PMID- 9724474 TI - Payer status, but not race, affects the cost of liver transplantation. AB - Prior studies evaluating the impact of race and payer on cost of liver transplantation did not adjust for clinical factors known to increase cost. We analyzed the impact of race and payer on the cost of liver transplantation after controlling for clinical factors. We analyzed data obtained on patient and graft survival, cost, race, age, sex, payer, and United Network for Organ Sharing (UNOS) status from 153 consecutive liver transplants in 130 patients performed at University of North Carolina Hospitals from September 1991 through December 1996. Race was classified as white or nonwhite, and payer status was classified as commercial or Medicare/Medicaid. Multivariate linear regression was used to compare costs, adjusting for age, sex, race, payer, and UNOS status. For the 130 patients, 1-year patient and graft survival rates were 88% and 82%, respectively. There were no significant differences in patient and graft survival or in the unadjusted average cost of liver transplantation by race or payer. After adjusting for demographic and clinical factors, the cost of transplantation was $28,494 more for Medicare/Medicaid recipients compared with the commercial insurance recipients (P = .02). The Medicare/Medicaid group had higher intensive care unit costs compared with the commercial insurance group ($17,807 and $9,359, respectively; P = .03), and a longer length of stay (41 and 31 days, respectively; P = .04). There was no significant difference in cost between whites and nonwhites adjusting for these factors. Medicare or Medicaid patients had a higher cost of transplantation compared with those with commercial insurance. The cost of liver transplantation was similar for whites and nonwhites. PMID- 9724475 TI - Parental psychosocial outcomes in pediatric liver and/or intestinal transplantation: pretransplantation and the early postoperative period. AB - Although liver transplantation has become an effective treatment for end-stage liver disease and liver/intestine transplantation is becoming an increasingly viable procedure for end-stage short-gut syndrome in children, little is known about the impact of these procedures on the child's family. Examination of the impact of these transplantations on the family is needed to identify psychosocial factors that may adversely affect the child's physical and emotional health and to plan for preventive interventions. The psychosocial impact of pediatric liver and/or intestine transplantation on parents was assessed in a cross-sectional sample of 41 mothers and 20 fathers evaluated pretransplantation and 2 months posttransplantation. Parental mental health, parenting stress, and quality of life were assessed, as well as demographic, child, and family characteristics as they related to parental outcomes. Parent adjustment did not differ with regard to time of assessment. A majority of parents (n = 31) reported elevated psychological symptoms on the Brief Symptom Inventory (BSI), with fathers showing greater distress than mothers (P < .05). Parents' total scores on the Parenting Stress Index and the Parent and Child Domain subscales were in the normal range. Quality of life was assessed by the Physical Health, General Health Perception, and Vitality subscales of the SF-36. Parents reported better physical functioning (P = .02) but lower vitality ratings than a normative population (P < .01). Family conflict was associated with higher psychological distress on the BSI (P = .02), whereas demographic factors, including the child's age, sex, and number of people in the household, proved most useful in predicting parenting stress and quality of life. These findings of significant psychological distress in parents of children undergoing liver and/or small-intestine transplantation have implications for the child's adaptation and underscore the need for psychosocial assessment of both parents in the perioperative period. Early identification of families at psychosocial risk and the development of interventions that may prevent or reduce psychological distress are indicated to ensure the best possible outcomes for these children and their families. PMID- 9724476 TI - Management of liver adenomatosis: results with a conservative surgical approach. AB - Liver adenomatosis is defined by the presence of multiple hepatic adenomas (more than three lesions). The natural history and treatment of liver adenomatosis are not yet well defined. The Mayo Clinic (Rochester, MN) experience with liver adenomatosis in the past 11 years was reviewed and a rational treatment approach is presented. Records from patients with liver adenomatosis and hepatic adenoma seen at the Mayo Clinic from January 1986 to June 1997 were reviewed. Estrogen- and progesterone-receptor status was assessed by immunohistochemistry. Eight women with liver adenomatosis were identified. All patients had undergone surgical treatment. Abdominal pain was the presenting symptom in 87.5% of the patients with adenomatosis and in 42.1% of the patients with hepatic adenoma. Tumor bleeding was present in 62.5% of the patients with adenomatosis and in 26.3% of the patients with hepatic adenomas. Bleeding occurred predominantly in lesions greater than 4 cm. All patients with liver adenomatosis reported improvement of symptoms after surgery, and the mean bleeding-free period after resection in 5 patients was 52.6 +/- 23.6 months. In 6 patients, estrogen receptor-positive and estrogen receptor-negative tumors were identified in the same liver. Based on the good outcome after resection in symptomatic patients with liver adenomatosis, we recommend resection of large (>/=5 cm) or symptomatic lesions with observation of smaller lesions (65 years was associated with early mortality after TIPS placement, but this trend was not statistically significant. All 4 subjects undergoing liver transplantation required perioperative pleural fluid drainage, but only 1 subject has experienced recurrent effusion. We conclude that TIPS may be a safe and effective temporizing treatment for carefully selected patients with refractory hepatic hydrothorax. However, patient survival is limited after TIPS and is primarily determined by availability of liver transplantation. PMID- 9724481 TI - Treatment of fulminant hepatic failure with intravenous prostaglandin E1. AB - Fulminant hepatic failure (FHF) is a severe, life-threatening disorder. Previous studies have suggested that intravenous prostaglandin treatment may improve survival in FHF. The present study was performed to further investigate the possible benefit of intravenous prostaglandin E1 (PGE1) for patients with FHF. A total of 18 patients, all excluded as candidates for hepatic transplantation, were studied. Thirteen of 18 participated in a randomized, double-blind, placebo controlled trial. PGE1 was administered by continuous infusion at a dose of 10 to 40 microg/h as tolerated. After 48 hours of blinded treatment, 3 of 7 patients randomized to placebo were converted to open-label PGE1 for lack of biochemical and/or clinical improvement. Mean values for alanine transaminase, aspartate transaminase, total bilirubin, prothrombin time, factor V percent, factor VII percent, hepatic encephalopathy score, days from onset of symptoms to initiation of treatment, and cause of FHF were similar between treatment groups. Ten of 18 patients (55%) enrolled in this trial survived. However, survival was not different between PGE1-(60%) and placebo (50%) treated patients. The greatest predictor of survival was the number of days from onset of symptoms to hospitalization, which was significantly (P = .002) shorter for survivors (3.3 v 12.4 days), regardless of PGE1 treatment. Six of 8 patients (75%) who began PGE1 therapy and 4 of 5 placebo-treated patients (80%) hospitalized within 10 days of onset of symptoms survived. By contrast, all 5 patients who were hospitalized and subsequently began PGE1 treatment 10 days or longer after the onset of symptoms died. We conclude that early recognition and hospitalization is the most important factor in reduction of mortality from FHF. It is unclear whether PGE1 treatment is beneficial when administered during this period. However, it is apparent that PGE1 was not effective for treatment of FHF if treatment started more than 10 days after onset of this clinical syndrome. PMID- 9724482 TI - Pharmacokinetics of tacrolimus and cyclosporine in short-bowel syndrome. PMID- 9724483 TI - Mycotic aneurysm of arterial conduit presenting as massive upper gastrointestinal hemorrhage after liver transplantation. PMID- 9724484 TI - De novo hepatitis B infection after liver transplantation: are all surgeons vaccinated against hepatitis B? PMID- 9724485 TI - Announcements and meetings PMID- 9724486 TI - Implications of the diabetes control and complications trial for renal outcomes and medical nutrition therapy. AB - The Diabetes Control and Complications Trial (DCCT) results have important implications related to the prevention and management of diabetic nephropathy. This paper provides an overview of the study design, methods and overall results with particular emphasis on renal outcomes and the role of medical nutrition therapy. The DCCT demonstrated that intensive diabetes therapy aimed at near normoglycemia resulted in a 39% reduction in the occurrence of microalbuminuria and a 54% reduction in the occurrence of albuminaria. Further analysis of DCCT data show that the impact of intensive therapy on renal outcomes may vary based on the stage of diabetic nephropathy, gender and prior exposure to hyperglycemia. This review highlights the clinical implications of the DCCT results for dietitians, nurses and physicians working with patients with diabetic nephropathy and describes post-DCCT advances in clinical practice and research. PMID- 9724487 TI - Effect of potassium concentration in dialysate on total body potassium. AB - OBJECTIVE: To investigate the effects of the presence/absence of potassium in the dialysate on total body potassium content in stable hemodialysis patients. DESIGN: Randomized selection. SETTING: Outpatient chronic hemodialysis unit at the James A. Haley VA Hospital in Tampa, Florida. PATIENTS: Six adult hemodialysis patients (mean age 48 +/- 11.61 years, range 32-65 years) participated in this study. They were all males, four African-Americans and two Caucasians. INTERVENTION: Subjects in a random order received dialysis using a dialysate containing no potassium for 3 months, followed by a 2 mEq/L of potassium for another 3 months. Total body potassium measurements and routine blood analysis were taken at baseline and after each three month period. All subjects received diet instruction pre and monthly on a 3 g/day potassium dietary restriction. MAIN OUTCOME MEASURED: There were no significant differences in the amount of total body potassium concentration between the two different dialysates containing 0 or 2 mEq/L potassium. Mean values of serum albumin were significantly higher when the subjects were dialyzed on a 0 potassium bath. CONCLUSION: The use of dialysate containing 0 or 2 mEq/L potassium concentration does not make a significant difference with regard to total body potassium concentration and maybe advantageous for the patients in terms of more freedom in nutritional intake as demonstrated by a significant increase in serum albumin when they were dialyzed on a 0 potassium bath. PMID- 9724488 TI - Change in nutritional status of patients on peritoneal dialysis. AB - OBJECTIVE: To document the prevalence of undernutrition/overnutrition in patients on peritoneal dialysis (PD) and to examine whether nutritional status (NS) changes with time on this form of dialysis. DESIGN: Retrospective observational study. Patients had been on PD >2 years. Data included age, gender, diagnosis, peritonitis rate, anthropometry and biochemistry. A classification system for NS was devised using BMI, TSF, MAMC and serum albumin. SETTING: Regional Peritoneal Dialysis Programme, University Teaching Hospital. PATIENTS: 82 patients were on PD on March 1994. A cohort of 28 patients remained on PD after 2 years and complete nutritional data was available for 23 of these patients (9 male, 14 female: mean age 58yrs). RESULTS: 65% of patients were classified as having an acceptable NS at the start of PD and 56% were classified as acceptable at the latest assessment. The prevalence of mild/moderate undernutrition both at the start of PD and at the latest assessment was 26% (different patients at each assessment). No patients were classified as severely undernourished. The prevalence of overnutrition at the start of PD was 9% and at the latest assessment was 17%. There was no statistically significant difference in NS between diabetics and non-diabetics nor between male and female patients although undernutrition was more frequently observed in males. Overnutrition increased with time in both genders but this did not reach statistical significance. There was no difference in initial NS between those who remained on PD and those who died. Change in NS was not related to peritonitis rate. CONCLUSION: Whereas this study has insufficient statistical power to avoid a Type II error it supports our clinical observation that NS does not substantially change with time in this population. There are, however, a small number of individuals who exhibit changes in NS. Given the difficulty in predicting change in NS with time, regular nutritional assessment is important to identify those who require more intensive dietetic intervention. PMID- 9724489 TI - Relationship between serum phosphorus levels and various outcome measures in adult hemodialysis patients. AB - OBJECTIVE: To compare three different mean serum phosphorus ranges on outcomes related to the control and treatment of hyperparathyroidism (HPTH), to nutritional status, and to quality of life (QOL) in adult hemodialysis (HD) patients. DESIGN: Patients were grouped based on the mean of five monthly phosphorus levels achieved during the study period. Group 1 included patients whose mean phosphorus levels over the period was <6.0 mg/dL (n = 24); group 2 averaged between 6.0 and 6.9 mg/dL (n = 14); and group 3 averaged >7.0 mg/dL (n = 16). Descriptive comparisons were made between phosphorus groups. PATIENTS: Fifty four stable, adult HD patients participated voluntarily. MAIN OUTCOME MEASURES: Intact-parathyroid hormone (iPTH), calcium x phosphorus product (Ca x P), and change in iPTH, albumin (alb), total protein (tpro), weight (wt) and body mass index (BMI), and scores on a QOL survey. Baseline physical and lab characteristics. RESULTS: No difference was found between phosphorus levels of <6.0 mg/dL and levels of 6.0 to 6.9 mg/dL in iPTH, Ca x P levels allowing safe calcitriol therapy, nor response to calcitriol treatment. Patients with phosphorus levels >7.0 mg/dL had midstudy iPTH greater than phosphorus levels <6.0 mg/dL. Otherwise the three groups did not differ significantly in iPTH levels. Phosphorus levels 6.0 to 6.9 mg/dL was associated with lowest wt and BMI, but alb and tpro did not differ between the phosphorus groups. Phosphorus levels of >7.0 was associated with highest creatinine levels and youngest age. Subjects in the phosphorus levels of <6.0 mg/dL gp were more likely than the 6.0 to 6.9 mg/dL gp to describe their diet as sufficient and, at baseline, were more likely to relate diet to QOL. CONCLUSION: Comparison of three levels of serum phosphorus on indicators of outcome in the control and treatment of secondary hyperparathyroidism showed no significant difference in outcome between phosphorus levels of <6.0 mg/dL and phosphorus levels 6.0 to 6.9 mg/dL. However, the data suggests that phosphorus levels of >7.0 mg/dL may relate to significantly higher iPTH and unacceptable Ca x P levels. There were no differences between the groups, suggesting less favorable outcome at any of the three phosphorus levels regarding nutritional status or QOL in this small group of stable, adult HD patients. PMID- 9724490 TI - The conservative management of chronic renal failure: results of a recent survey. AB - Opinions differ regarding the role of protein restriction in the progression of renal disease. Unfortunately the Modification of Diet in Renal Disease Study did not settle this controversy. To assess the use of low protein diets (LPDs) in the United Kingdom, renal dietitians were surveyed about the current dietetic practices and beliefs in their Renal Units. In the survey, 81% of questionnaires were returned (59 of 73), of these 35 Renal Units were using LPDs (<1 g/kg body weight). The majority (33) used LPDs for symptomatic relief and 20 for slowing progression. The use of LPDs in diabetic patients was similar to nondiabetic patients, but LPDs were started earlier by 12 Renal Units. Most (22) of the dietitians calculate LPDs on the basis of ideal body weight. Nutritional assessment was by weight (35), body mass index (26), serum albumin (31), and upper arm anthropometry (10). The lack of agreement, both within and outside of Renal Units will ensure that the debate regarding protein restriction continues. PMID- 9724491 TI - The dynamics of education: making a match. AB - The education process when applied to the renal population offers challenges to the health educator. Complexity of information, the numerous and often changing diet restrictions, along with the learning abilities of clients present potential obstacles to the education process. Selecting materials that match the client's learning abilities can enhance learning outcomes. The various methods to assess teaching materials are presented along with guidelines for the development of educational tools. PMID- 9724492 TI - Renal DETERMINE nutrition screening tools for the identification and treatment of malnutrition. AB - Nutrition screening is the first step in identifying and treating nutrition related problems in renal patients. The Renal DETERMINE Nutrition Screening Tools help health care professionals recognize the risk factors for malnutrition in renal patients and suggest interventions to prevent, control, or ameliorate problems when they are present. The Renal DETERMINE Nutrition Awareness Checklist provides a series of questions for the health care professional to ask the renal patient to better identify nutrition problems. It can also be used with renal patients to help educate and increase awareness of nutrition issues. The Renal DETERMINE Nutrition Screening Reference Sheets are then used to help the health care professional identify appropriate interventions for the nutrition problem. The Reference Sheets list the most common nutrition-related concerns for chronic renal insufficiency, hemodialysis, peritoneal dialysis, and post kidney transplant patients. For each risk factor, rationales are presented and interventions to resolve the nutrition related problems are provided. PMID- 9724493 TI - Ideas for treating low blood sugar for diabetics on dialysis. PMID- 9724494 TI - Abstracts from the seventh annual nfk spring clinical nephrology meetings march 26-29, 1998 nashville, tennessee PMID- 9724496 TI - Call for abstracts PMID- 9724495 TI - Message from the chairperson PMID- 9724497 TI - 9(th) international congress on nutrition and metabolism in renal disease PMID- 9724498 TI - Medicare policy concerning nutritional therapy for end stage renal disease patients. PMID- 9724499 TI - Levocarnitine and muscle metabolism in patients with end-stage renal disease. AB - Levocarnitine is a molecule required in mammalian energy metabolism. It removes the potentially toxic acyl groups from the cell helping to maintain normal metabolic functions. In addition, it facilitates the transport of long-chain fatty acids across the mitochondrial membrane for beta oxidation and subsequent energy production in skeletal muscle and myocardium. It has been shown in numerous studies that levocarnitine metabolism is abnormal in patients with end stage renal disease. Significant dialytic loss of levocarnitine has been reported in addition to dietary changes undertaken in this population, which may decrease dietary levocarnitine intake. Recent studies have shown that levocarnitine administration to hemodialysis patients has improved exercise performance, intradialytic muscle cramps and hypotension episodes, and overall well-being. Ongoing and future studies will help to formulate more definite recommendations on the dose and the duration of levocarnitine therapy in dialysis patients. PMID- 9724500 TI - Nutritional intervention and growth in children with chronic renal failure. AB - OBJECTIVE: To assess whether improving energy intake by tube feeding could prevent growth failure and improve growth rates in children with congenital renal failure. DESIGN: Prospective descriptive study. SETTING: Renal Units, Royal Alexandra Hospital for Children, and Westmead Hospitals. PATIENTS: All children with advanced chronic renal disease (glomerular filtration rate < 30 mL/min/1.73 m2) between 1992 and 1994. INTERVENTION: Tube feeding was commenced if height or weight standard deviation score (SDS) was below the normal range (> -2 SDS) or when height SDS was decreasing and oral intake was not meeting energy requirements. Energy requirements were calculated for median weight for chronological age and sex to provide for catch-up growth. MAIN OUTCOME MEASURES: Growth rate was measured by comparing height and weight SDS at the beginning and end of the study period. Normal growth rate is defined as no change in SDS over time, whereas catch-up growth is defined as an increase in SDS over time. RESULTS: Seven children, mean age 0.6 +/- 0.7 years, with advanced renal failure (mean glomerular filtration rate = 17 mL/min/1.73 m2) caused by congenital renal hypoplasia/dysplasia were studied. All subjects were eventually tube fed for a mean time of 18. 6 +/- 4.5 months. There was no significant change in height SDS (-0. 9 to -1.1) or weight SDS (-0.4 to -0.2). CONCLUSION: Optimizing nutritional intake by tube feeding children with advanced chronic renal failure from an early age resulted in no decline in growth rate; however, catch-up growth was not achieved. PMID- 9724501 TI - Potassium and sodium intake and excretion in calcium stone forming patients. AB - OBJECTIVE: To determine mean potassium (K) intake and its correlation with urinary calcium (uCa) and citrate excretion, as well as uCa, sodium (Na), and K levels of calcium stone forming patients. We determined the K-rich foods most commonly consumed by these patients. DESIGN: Case-control. SETTING: University affiliated outpatient renal Lithiasis Unit. PATIENTS AND CONTROLS: One hundred hypercalciuric calcium stone forming patients (CSF, 54 men/46 women), 37 with associated hypocitraturia, were sequentially enrolled in the study that was performed before the initiation of any care for their renal stones. The control group consisted of 100 age-matched healthy subjects (HS, 47 men/53 women) who were laboratory employees with no history of renal stones. INTERVENTION: The analyses consisted of a 3-day dietary record to determine the mean K and calcium (Ca) intakes, and a 24-hour urine sample with measurements of K, Ca, Na, and citrate. MAIN OUTCOME MEASURE: K and Na intake determined by dietary record. RESULTS: uCa and Na levels and the Na/K ratio were significantly higher for CSF versus HS (238 +/- 118 v 148 +/- 74 mg/24 hours, 238 +/- 100 v 181 +/- 68 mEq/24 hours, 6.6 +/- 3.5 v 5.1 +/- 2.3, respectively, P < .05). The mean citrate excretion was lower in CSF than in HS patients (410 +/- 265 v 530 +/- 240 mg/24 hours). Mean uCa did not differ between groups. CSF patients showed a higher sodium chloride intake compared with HS (14 +/- 4 vs 8 +/- 3 g/day). The mean Ca intake of CSF and HS were 559 +/- 327 and 457 +/- 363 mg/day, respectively. The mean K intake of CSF and HS were 58 +/- 17 and 51 +/- 27 mEq/day. A positive correlation was observed between uCa and urinary sodium (r = .40 and r = .65, P < .05), urinary potassium and urinary citrate (r = .25 and r = .53, P < .05), uCa and Na/K (r = .33 and r = .56, P < .05) respectively for CSF and HS. The following were the K-rich foods consumed at least once a day by these groups: beans (by 70% of CSF and 75% of HS), tomatoes (by 42% of CSF and 50% of HS), oranges (by 30% of CSF and 55% of HS), and bananas (by 42% of CSF and 23% of HS). CONCLUSION: Despite the consumption of K-rich foods at least once a day, the mean K intake by CSF patients was 58 mEq/day. This intake can still be considered to be low, although it meets recommended daily dietary allowance requirements. Therefore, we describe herein a population of CSF with high-Na intake and normal- to low-K intake, which may contribute to stone formation. PMID- 9724502 TI - Percentage body fat determination in hemodialysis and peritoneal dialysis patients: a comparison. AB - OBJECTIVE: To evaluate percentage body fat in hemodialysis (HD) and peritoneal dialysis (PD) patients. DESIGN: A prospective study of 20 HD patients and 20 PD patients. SETTING: Sol Goldman Renal Therapy Center, Lenox Hill Hospital, New York, NY; Baumritter Kidney Center Albert Einstein College of Medicine, Bronx, NY; Body Composition Unit, St Luke's Roosevelt Hospital, Columbia University, New York, NY. PATIENTS: Twenty HD (10 men, 10 women) patients, mean age 41.8 +/- 2.4 years and 20 PD (12 men, 8 women) patients, mean age 48.6 years +/- 3.0 years. INTERVENTION: This is a noninterventional study. PATIENTS signed consent to undergo dual-energy x-ray absorptiometry, total body potassium counting bioelectrical impedance analysis, total body water determination, and anthropmetric evaluation. MAIN OUTCOME MEASURES: Present and compare percentage body fat between HD and PD patients as determined by the methods used. RESULTS: Percentage fat is not different between HD and PD patients. Differences in absolute values of percent fat between techniques exist. CONCLUSION: HD patients and PD patients may be evaluated by the methods of body composition used. Percentage body fat will vary among techniques; therefore the same method should be used to follow a patient over time. PMID- 9724503 TI - The effect of dietary intervention on weight gains after renal transplantation. AB - OBJECTIVES: To determine the effect of early intensive dietary intervention and follow-up on weight gains in newly transplanted renal patients. To provide appropriate dietary advice posttransplant that included advice to reduce weight gains. DESIGN: Group A was studied prospectively and group B was studied retrospectively over a period of 1 year posttransplant. SETTING: Hospital transplant unit: inpatient ward and outpatient clinic. PATIENTS: Thirty-three transplant patients were studied: Group A consisted of 11 patients (9 men, 2 women) transplanted consecutively over 2 months, with a mean age of 39 years. Group B consisted of 22 patients (14 men, 8 women) who had been transplanted consecutively 4 years before the study, with a mean age of 40 years. Both groups had functioning grafts (serum creatinine <200 micromol/L [2.2 mg/dL]) over the study period, and similar triple immunosuppressive therapy (prednisolone, cyclosporine, and azathioprine). INTERVENTION: Group A received intensive, individualized dietary advice in stages, with regular follow-up for the first 4 months posttransplant. Thereafter group A did not receive any dietary advice or follow-up for the 8 months leading up to 1 year posttransplant. Group B had not received any dietary advice or follow-up posttransplant. MAIN OUTCOME MEASURE: Weight gained and body mass index (BMI) at 4 months and at 1 year posttransplant. RESULTS: The mean weight (BMI) for group A at baseline, 4 months and at 1 year posttransplant was 67 +/- 13 kgs (24.1 +/- 3.9 kg/m2), 69 +/- 12 kgs (24.6 +/- 3.5 kg/m2), and 73 +/- 12 kgs (26.1 +/- 3.4 kg/m2), respectively. The mean weight (BMI) for group B at baseline, 4 months and at 1 year posttransplant were 67 +/- 11 kgs (23.7 +/- 3.4 kg/m2), 74 +/- 9 kgs (26.3 +/- 3.3 kg/m2), and 79 +/- 12 kgs (27.9 +/- 4 kg/m2), respectively. Analysis of group A showed no significant difference in weight gained and BMI with dietary advice and follow-up at 4 months posttransplant compared with baseline. There was a significant difference in weight gain and BMI at 1 year posttransplant compared with 4 months posttransplant (P = .002, P = .002, respectively). Analysis between groups showed a significantly lower weight gain in group A compared with group B both at 4 months and at 1 year posttransplant (P = .01, P = .01 respectively). Group A had a significantly lower BMI than group B at 4 months and at 1 year posttransplant (P = .003, .006, respectively). At 1 year posttransplant, group A had a mean weight gain of 5.5 kg per patient compared with a mean of 11.8 kg per patient in group B. CONCLUSION: Early intensive dietary advice and follow-up is effective in controlling weight gains in the first year posttransplant. Dietary advice should be an important part of posttransplant treatment. PMID- 9724504 TI - Validation of serum transthyretin (prealbumin) as a nutritional parameter in hemodialysis patients. AB - OBJECTIVE: To evaluate the use of serum transthyretin (TTR) as a valid indicator of nutritional status in the hemodialysis patient and to validate the correlation of low-serum (TTR) levels with established nutrition assessment parameters. DESIGN: Prospective, cohort, correlation analysis. SETTING: Free-standing outpatient dialysis center. PATIENTS: Fifty-one stable, chronic hemodialysis patients meeting the following selection criteria: (1) received thrice weekly hemodialysis treatments for greater than 3 months, (2) absence of impaired hepatic function, (3) absence of chronic infection, inflammatory syndromes, or infections in the 3 months before the study, (4) not taking corticosteroids, and (5) willing to participate in the study as evidenced by signing of an informed consent. INTERVENTION: Serum TTR, albumin, blood urea nitrogen, creatinine, cholesterol, postdialysis weight and body mass index were measured monthly for 6 consecutive months. Normalized protein catabolic rate and KT/V were measured monthly for 3 consecutive months. MAIN OUTCOME MEASURES: Nutrition and biochemical indices. RESULTS: The overall mean TTR level was 32 mg/dL +/- 7 for the 6-month study period. Thirty-six percent of patients had mean TTR levels less than 30 mg/dL. TTR levels less than 30 mg/dL correlated significantly with urine outputs greater than 240 mL/24 hours, predialysis blood urea nitrogen < 18 mmol/L (<50 mg/dL), and normalized protein catabolic rate less than 0.8 g/kg/d (P < .05). A significant correlation was found between TTR and creatinine, albumin and loss of dry body weight (P < .05). Mean TTR levels less than 30 mg/dL were found in 33% of subjects with mean albumin levels greater than 35 g/L (>3.5 g/dL) and in 19% with mean albumin levels greater than 40 g/L (>4.0 g/dL). TTR levels were consistently lower in diabetics for all 6 months (statistically significant in 2 out of 6 months). CONCLUSION: Measuring serial serum TTR levels in hemodialysis patients is a reliable method for identifying patients in need of nutrition intervention. PMID- 9724505 TI - Nutritional control of pregnant women on chronic hemodialysis. AB - The authors describe their experience in the follow-up of four patients with chronic renal failure who became pregnant while being treated with chronic hemodialysis. The outcomes were successful and each gave birth to healthy babies. The adequate nutritional condition previous to the pregnancies added more safety to their management. Special dedication to the nutritional control enabled a good outcome of their pregnancies. It stressed the importance of the intervention of the nutritionist-dietitian in the follow-up of nephrologic patients and the integration of a multidisciplinary staff. PMID- 9724506 TI - Reducing cholesterol by diet for dialysis patients. PMID- 9724508 TI - Message from the chairperson PMID- 9724509 TI - CALL FOR ABSTRACTS. PMID- 9724510 TI - Crystal structure of an eight-base pair duplex containing the 3'-DNA-RNA-5' junction formed during initiation of minus-strand synthesis of HIV replication. AB - During initiation of minus-strand synthesis by HIV-1 reverse transcriptase, a 3' DNA-RNA-5' junction is formed involving the 3'-end of tRNAlys,3. The HIV-RT associated RNase H cleaves the RNA template strand specifically, opposite the newly synthesized DNA strand. We have determined the crystal structure at 1.9 A resolution of an eight-base pair hybrid duplex representing the junction to identify global or local structural perturbations which may be recognized by HIV RT RNase H. The junction octamer is in a global A-type conformation throughout. A base pair step with distinct stacking geometry and variable backbone conformation is located next to the main endonucleolytic cleavage site. This base pair step may serve as a recognition site for HIV-RT RNase H. PMID- 9724511 TI - On the mechanism of 5-enolpyruvylshikimate-3-phosphate synthase. AB - 5-Enolpyruvylshikimate-3-phosphate (EPSP) synthase catalyzes the condensation of shikimate 3-phosphate (S3P) and phosphoenolpyruvate (PEP) to form EPSP, a precursor for the aromatic amino acids. This paper examines a recent claim [Studelska, D. R., McDowell, L. M., Espe, M. P., Klug, C. A., and Schaefer, J. (1997) Biochemistry 36, 15555-15560] that the mechanism of EPSP synthase involves two covalent enzyme-intermediates, in complete contrast to a large body of literature that has already proven the involvement of a single noncovalent intermediate. The evidence in the paper of Studelska et al. is examined closely, and unequivocal proof is provided that those authors' NMR assignments to covalent structures are in error, and that in fact the species they observed were simply the product EPSP and a side-product EPSP ketal. Since those authors used rotational-echo double-resonance (REDOR) solid-state NMR to measure intermolecular and intramolecular distances in the proposed covalent intermediates, we have used REDOR to measure the same distances in enzyme-free and enzyme-bound preparations of purified EPSP, and enzyme-free preparations of purified EPSP ketal. The distance between the shikimate ring phosphorus atom and C8 in enzyme-free EPSP is 6.6 +/- 0.1 A, which lengthens to 7.4 +/- 0.1 A in the presence of the enzyme, and in enzyme-free EPSP ketal is 5.6 +/- 0.1 A. These are entirely consistent with those measured by Studelska et al., which were 7.5 +/- 0.5 A for a putative enzyme-enolpyruvyl species and 6.1 +/- 0.3 A for a putative enzyme-ketal species. PMID- 9724512 TI - Lipid structure and not membrane structure is the major determinant in the regulation of protein kinase C by phosphatidylserine. AB - This study addresses the molecular basis for protein kinase C's specific activation by phosphatidylserine. Specifically, we ask whether protein kinase C's phospholipid specificity arises from specific protein/lipid interactions or whether it arises from unique membrane-structuring properties of phosphatidylserine. We measured the interaction of protein kinase C betaII to membranes that differed only in being enantiomers to one another: physical properties such as acyl chain composition, membrane fluidity, surface curvature, microdomains, headgroup packing, and H-bonding with water were identical. Binding and activity measurements reveal that protein kinase C specifically recognizes 1, 2-sn-phosphatidyl-L-serine, independently of membrane structure. High-affinity binding and activation are abolished in the presence of enantiomeric membranes containing 2,3-sn-phosphatidyl-L-serine, 2, 3-sn-diacylglycerol, and 2,3-sn phosphatidylcholine. Our data also show that the stereoselectivity for 1,2-sn diacylglycerol is not absolute; 2,3-sn-diacylglycerol modestly increases the membrane affinity of protein kinase C provided that 1, 2-sn-phosphatidyl-L-serine is present. We also find that the stereochemistry of the bulk phospholipid, in this case phosphatidylcholine, has no significant influence on protein kinase C's membrane interaction. These data reveal that specific molecular determinants on protein kinase C stereospecifically recognize structural determinants of phosphatidylserine. PMID- 9724513 TI - Comprehensive DNA recognition through concerted interactions from adjacent zinc fingers. AB - Zinc fingers are small DNA-binding modules noted for their occurrence in a large number of eukaryotic transcription factors, and their use in protein engineering. Although it was expected that zinc fingers can bind to a wide diversity of DNA sequences, previous studies using model zinc finger domains from Zif268 (and Sp1) have revealed a potential limitation to the DNA-binding specificity. For example, phage display selection of individual zinc fingers to recognize trinucleotide DNA subsites returned fingers that bound specifically only to triplets of the form GNN, i.e., triplets with guanine at the 5' end. Following our recently reported work [Isalan, M., Choo, Y., and Klug, A. (1997) Proc. Natl. Acad. Sci. U.S.A. 94, 5617-5621], we now show that this limitation can be overcome by the concerted randomization of certain amino acid positions in adjacent zinc fingers that specify overlapping DNA subsites. This illustrates an important mechanism underlying DNA recognition by arrays of zinc fingers, and points the way to improved strategies for the design of highly specific zinc finger proteins that bind any given nucleotide sequence. PMID- 9724514 TI - Mutagenic specificity of (acetylamino)fluorene-derived DNA adducts in mammalian cells. AB - Site-specifically modified oligodeoxynucleotides were used to explore the mutagenic potential of dG-AAF and dG-AF adducts in mammalian cells. The miscoding properties of these arylamine adducts were established by analyzing fully extended products of primer extension reactions catalyzed by mammalian DNA polymerases alpha, beta, and delta. On DNA templates containing dG-AAF, pol alpha generated two-base deletions and promoted incorporation of small amounts of dCMP, dAMP, and dTMP opposite the lesion. Reactions with pol beta were associated exclusively with two-base deletions. Primer extension catalyzed by pol delta was strongly blocked by the adduct. On DNA templates containing dG-AF, all three DNA polymerases generated full-length products, preferentially incorporating dCMP opposite the lesion. A single-stranded shuttle vector containing 5'TCCTCCTCXCCTCTC (X = dG-AAF, dG-AF, or dG) was used to establish the frequency and specificity of dG-AAF- and dG-AF-induced mutations in simian kidney (COS-7) cells. Vectors containing a single dG-AAF or dG-AF adduct promote significant incorporation of dAMP and lesser amounts of dTMP opposite the lesion. dG-AAF also promoted some incorporation of dGMP and a two-base deletion. dG-AAF was 3.8 times more mutagenic than dG-AF (11% vs 2.9%) in COS cells. We conclude from this study that dG-AAF and dG-AF produce G --> T transversions and, to a much lesser degree, G --> A transitions in mammalian cells. PMID- 9724515 TI - Tryptophan substitutions surrounding the nucleotide in catalytic sites of F1 ATPase. AB - Novel tryptophan substitutions, surrounding the nucleotide bound in catalytic sites, were introduced into Escherichia coli F1-ATPase. The mutant enzymes were purified and studied by fluorescence spectroscopy. One cluster of Trp substitutions, consisting of beta-Trp-404, beta-Trp-410, beta-Asp-158 (lining the adenine-binding pocket), and beta-Trp-153 (close to the alpha/beta-phosphates), showed the same fluorescence responses to MgADP, MgAMPPNP, and MgATP and the same nucleotide binding pattern with MgADP and MgAMPPNP, with one site of higher and two sites of lower affinity. Therefore, in absence of catalytic turnover (and of gamma-subunit rotation), sites 2 and 3 appeared similar in affinity, and the region of the catalytic site sensed by these Trp substitutions did not change conformation with different nucleotides. In contrast, alpha-Trp-291 and beta-Trp 297, both close to the gamma-phosphate, showed very different fluorescence responses to MgADP versus MgAMPPNP, and in these cases the response was due exclusively or predominantly to nucleotide binding at the first, high-affinity catalytic site, thus allowing specific detection of this site. Titration with MgATP showed that the high-affinity site was present under conditions of steady state, Vmax MgATP hydrolysis. PMID- 9724516 TI - Binding of quinacrine to acidic phospholipids and pancreatic phospholipase A2. Effects on the catalytic activity of the enzyme. AB - Binding of quinacrine to phospholipids and porcine pancreatic phospholipase A2 (PLA2) was investigated using fluorescence resonance energy transfer, Langmuir films, assay for the enzymatic activity, and molecular modeling. No significant binding of this drug to the zwitterionic phosphatidylcholine was observed whereas a high affinity for acidic phospholipids was revealed by quenching of pyrene labeled phospholipid analogues. Partial reversal of this binding was observed due to the addition of 4 mM CaCl2. Quinacrine efficiently and independently of the lipid surface pressure penetrated into monolayers of phosphatidylglycerol while only a weak penetration into phosphatidylcholine films was evident. Quinacrine also bound to eosin-labeled PLA2, and the addition of 4 mM CaCl2 reversed this interaction almost completely. In the presence of acidic phospholipids both the drug and the enzyme were attached to the lipid surface. Studies on the influence of quinacrine on the activity of PLA2 toward pyrene-labeled phospholipid analogues revealed that the hydrolysis of phosphatidylcholine was progressively reduced as a function of increasing [quinacrine]. At low [CaCl2] and low quinacrine:lipid molar ratios (<1:5) quinacrine enhanced slightly the rate of hydrolysis of acidic phospholipids whereas at higher drug:lipid molar ratios (>1:2) an inhibition was observed. In the presence of 1 mM CaCl2 quinacrine inhibited PLA2-catalyzed hydrolysis of phosphatidylglycerol only when the drug:lipid molar ratio exceeded 1:1. The presence of 4 mM CaCl2 abolished nearly completely the inhibition with all the substrate analogues used. Our data suggest that the inhibition of PLA2 by quinacrine is due to its binding to the enzyme. This is supported also by molecular modeling which suggested a binding site for quinacrine close to the active site and Ca2+ binding site of the enzyme. Importantly, our data indicate that quinacrine binds avidly to acidic phospholipids and their presence may influence the drug-enzyme interaction and the inhibition of the enzyme action. Accordingly, presence of quinacrine may interfere also with other processes that require the presence of acidic lipids and/or Ca2+, such as the function of the nicotinic acetylcholine receptor. PMID- 9724517 TI - Pro region C-terminus:protease active site interactions are critical in catalyzing the folding of alpha-lytic protease. AB - alpha-Lytic protease is encoded with a large (166 amino acid) N-terminal pro region that is required transiently both in vivo and in vitro for the correct folding of the protease domain [Silen, J. L. , and Agard, D. A. (1989) Nature 341, 462-464; Baker, D., et al. (1992) Nature 356, 263-265]. The pro region also acts as a potent inhibitor of the mature enzyme [Baker, D., et al. (1992) Proteins: Struct.,Funct., Genet. 12, 339-344]. This inhibition is mediated through direct steric occlusion of the active site by the C-terminal residues of the pro region [Sohl, J. L., et al. (1997) Biochemistry 36, 3894-3904]. Through mutagenesis and structure-function analyses we have begun to investigate the mechanism by which the pro region acts as a single turnover catalyst to facilitate folding of the mature protease. Of central interest has been mapping the interface between the pro region and the protease and identifying interactions critical for stabilizing the rate-limiting folding transition state. Progressive C-terminal deletions of the pro region were found to have drastic effects on the rate at which the pro region folds the protease but surprisingly little effect on inhibition of protease activity. The observed kinetic data strongly support a model in which the detailed interactions between the pro region C-terminus and the protease are remarkably similar to those of known substrate/inhibitor complexes. Further, mutation of two protease residues near the active site have significant effects on stabilization of the folding transition state (kcat) or in binding to the folding intermediate (KM). Our results suggest a model for the alpha-lytic protease pro region-mediated folding reaction that may be generally applicable to other pro region-dependent folding reactions. PMID- 9724518 TI - The chirality of phosphatidylserine and the activation of protein kinase C. AB - The properties of phosphatidyl-L-serine (L-PS) and phosphatidyl-D-serine (D-PS) were compared. The two forms of PS have similar but nonidentical L to L phase transition temperatures. Mixtures of phosphatidylserine with phosphatidylethanolamine and cholesterol (molar ratio 1:1:2) show polymorphic behavior at higher temperatures and in the presence of Ca2+. Mixtures with L-PS undergo conversion to nonlamellar phases at lower temperatures than do similar mixtures with D-PS. The aggregation of vesicles upon addition of histones is greater for L-PS than for D-PS. With fluorescence digital imaging microscopy we could show differences in the extent of formation of histone-induced domains enriched in PS or in diacylglycerol. The most enriched domains were induced with histone in membranes containing L-PS. The MARCKS peptide showed no differences in domain formation between L-PS and D-PS. The maximal activity of protein kinase C was greater in the presence of L-PS when histone, which could form more enriched domains, was the substrate. However, with a MARCKS peptide substrate, which formed domains of equal enrichment with L-PS and D-PS, the maximal activity of protein kinase C was the same with D-PS and with L-PS. These observations demonstrate that L-PS and D-PS have different physical properties. These differences likely contribute to the greater ability of L-PS to activate protein kinase C. PMID- 9724519 TI - Cysteine reactivity and oligomeric structures of phospholamban and its mutants. AB - To test models for the pentameric structure of phospholamban (PLB) and study its structure and molecular dynamics in SDS solution, we characterized recombinant PLB and several of its mutants by (a) reactivity of cysteine residues toward DTNB [5, 5'-dithiobis(2-nitrobenzoic acid)] and a thiol-reactive spin label, (b) oligomeric state on SDS-PAGE, and (c) EPR of the spin-labeled proteins. WT-PLB has three cysteine residues (36, 41, and 46), all located in the hydrophobic C terminal transmembrane region. In SDS at pH 7.5, exhaustive reaction with either sulfhydryl reagent resulted in essentially 2 mol of cysteine reacted/mol of WT PLB, with only slight destabilization of the native pentameric structure. When WT PLB was denatured in guanidine at pH 8.1, all three cysteines reacted, disrupting the pentamer, which was restored upon cleavage of the disulfide bonds with DTT. In the tetrameric mutant C41L-PLB, the two remaining cysteine residues reacted, reversibly destabilizing the tetramer. In the monomeric mutant L37A-PLB, all three cysteines reacted. The pentameric double cysteine replacement mutant C36,46A-PLB showed negligible reactivity. We conclude that Cys-41 is the unreactive cysteine in PLB and is located at a crucial site for the maintenance of the pentameric structure. EPR spectra in SDS of spin-labeled WT-PLB and mutants correlate with the oligomeric state on SDS-PAGE; oligomeric proteins show decreased spin-label mobility compared with monomers. Molecular dynamics calculations were used to construct an atomic model for the transmembrane region of the PLB pentamer, constrained by previous mutagenesis results and the results of the present study. We conclude that (a) the mobilities of spin-labels attached to PLB and its mutants are sensitive to oligomeric state and (b) the pattern of cysteine reactivity, spin-label mobility, and oligomeric state supports a structural model for the PLB pentamer in which interactions between each pair of subunits are stabilized by a leucine-isoleucine zipper. PMID- 9724520 TI - NMR solution structure of the N3' --> P5' phosphoramidate duplex d(CGCGAATTCGCG)2 by the iterative relaxation matrix approach. AB - High-resolution 2D NMR spectra of the duplex CGCGAATTCGCG with deoxyribose sugars but with the normal phosphodiester linker replaced by an N3' --> P5' phosphoramidate (NP) group have been used to establish a solution structure for the duplex. Distance, angle, and base pair hydrogen-bonding constraints were used to refine the structure by use of the iterative relaxation matrix approach (IRMA). The phosphoramidate NH proton signal could be observed in DMSO at low temperature but not in H2O and D2O. For this reason, the structure was refined with the -NH in each of the two possible low-energy configurations. The structure with the nitrogen lone pair located between the nonbridging oxygen atoms of the 5'-phosphate group consistently had the lowest energy and RMSD values, consistent with an X-ray analysis of the same duplex [Tereshko, V., Gryaznov, S. , and Egli, M. (1998) J. Am. Chem. Soc. 120, 269-283]. In the refined structure, the sugars are in the C3'-endo conformation with the change from the normal C2'-endo conformation of deoxyribose apparently being driven by the gauche effect and the change in electronegativity from the 3'O to the 3'NH group. In agreement with preliminary studies [Ding, D., Gryaznov, S. M., Lloyd, D. H., Chandrasekaran, S., Yao, S., Ratmeyer, L., Pan, Y., and Wilson, W. D. (1996) Nucleic Acids Res. 24, 354-360], the backbone conformation in the NP duplex is very close to classical A form values. Comparison of phosphodiester and phosphoramidate structures suggests that their backbones have global conformations that are primarily a function of the low-energy state of the sugar ring. A somewhat more complex situation arises when base pair conformation is analyzed with many of the base pairs having a conformation between those of classical A- and B-form helices. The effects of the 2' substituent are obviously important in specifying the final conformation of the stacked bases in either an A-form or B-form helix. It is clear, however, that conversion of the normal phosphodiester of DNA into a phosphoramidate linkage yields a nucleic acid that behaves much more like RNA than DNA, and it has been shown that NP sequences can bind to RNA-directed proteins [Rigl, C. T., Lloyd, D. H., Tsou, D. S., Gryaznov, S. M., and Wilson, W. D. (1997) Biochemistry 36, 650 659]. PMID- 9724521 TI - Highly selective mechanism-based thrombin inhibitors: structures of thrombin and trypsin inhibited with rigid peptidyl aldehydes. AB - The crystal structures of three highly potent and selective low-molecular weight rigid peptidyl aldehyde inhibitors complexed with thrombin have been determined and refined to R values 0.152-0. 170 at 1.8-2.1 A resolution. Since the selectivity of two of the inhibitors was >1600 with respect to trypsin, the structures of trypsin-inhibited complexes of these inhibitors were also determined (R = 0.142-0.157 at 1.9-2.1 A resolution). The selectivity appears to reside in the inability of a benzenesulfonamide group to bind at the equivalent of the D-enantiomorphic S3 site of thrombin, which may be related to the lack of a 60-insertion loop in trypsin. All the inhibitors have a novel lactam moiety at the P3 position, while the two with greatest trypsin selectivity have a guanidinopiperidyl group at the P1 position that binds in the S1 specificity site. Differences in the binding constants of these inhibitors are correlated with their interactions with thrombin and trypsin. The kinetics of inhibition vary from slow to fast with thrombin and are fast in all cases with trypsin. The kinetics are examined in terms of the slow formation of a stable transition-state complex in a two-step mechanism. The structures of both thrombin and trypsin complexes show similar well-defined transition states in the S1 site and at the electrophilic carbon atom and Ser195OG. The trypsin structures, however, suggest that the first step in a two-step kinetic mechanism may involve formation of a weak transition-state complex, rather than binding dominated by the P2-P4 positions. PMID- 9724522 TI - Preferential interaction of the mRNA proofreading factor TFIIS zinc ribbon with rU.dA base pairs correlates with its function. AB - The transcriptional factor TFIIS helps overcome elongation barriers and enhances proofreading by RNA polymerase II. These TFIIS functions may be modulated by the TFIIS zinc ribbon domain through interactions with nucleic acids in the elongation complex. Within this zinc ribbon domain, the dipeptide sequences Asp261-Glu262 and Arg276-Trp277 have been shown to be critical for its function by mutant analysis. The sequence Asp261-Glu262 has been suggested to participate in metal binding within the RNA polymerase II active site. We now show that the sequence Arg276-Trp277 interacts with nucleic acids through a combination of electrostatic and stacking interactions. The interaction of the indole side chain of the tryptophan residue with nucleic acid bases is demonstrated by a characteristic and reversible decrease in the zinc ribbon fluorescence intensity as a function of oligonucleotide concentration. These interactions are salt sensitive (maximum interaction at 200 mM and no interaction at 500 mM NaCl), suggesting that the tryptophan stacking with nucleic acid base accompanies electrostatic contacts. The oligonucleotide-zinc ribbon interactions exhibit small but significant base preferences, as shown by the dependence of Keq on base composition, with decreasing Keq in the order U > T > A > C >> G. Within the variety of homopolymeric single- and double-stranded deoxy- and ribooligonucleotides, the oligonucleotide rU12-18.dA20 exhibited a 2-6-fold binding preference relative to other oligonucleotides. This preferential binding of the zinc ribbon to sequences composed of rU.dA base pairs, which are generally associated with elongation blocks, may help in overcoming elongation barriers. Since the mRNA proofreading and enhancement of elongation involve cleavage of ribonucleotide of the mismatched pair and the weakly paired rU.dA nucleotides, but not the stably paired rC.dG nucleotides, we propose that the Arg276-Trp277 sequence in the TFIIS zinc ribbon may serve as a scanner connected to the transcript cleavage apparatus for weakly paired or mismatched nucleotides by employing indole ring stacking with the bases as a criterion of determining their subsequent removal. The striking similarity in preference for mismatched and weakly paired nucleotides for binding and for excision suggests a functional relationship between binding and cleavage reactions. PMID- 9724523 TI - Selective recognition and cleavage of RNA loop structures by Ni(II).Xaa-Gly-His metallopeptides. AB - The recognition and cleavage of tRNAPhe and the TAR RNA of HIV-1 by metallopeptides of the general form Ni(II).Xaa-Gly-His (where Xaa is Gly, Lys, or Arg) were investigated. The results of RNA cleavage analyses suggest that KHSO5- or magnesium monoperoxyphthalate-activated metallopeptides (1) induce nucleobase damage which requires aniline acetate for complete RNA strand scission and (2) selectively target the loops of stem-loop structures of the above-named substrates. In targeting RNA loop regions, the metallopeptides may be sensitive to intraloop structural features, including the overall structural environment of the loop itself and possibly the presence of intraloop hydrogen bonding. Overall, these results suggest that the metallopeptides interact selectively within a loop, in a fashion reminiscent of many RNA binding proteins, instead of targeting RNA single-stranded character alone. These observations further suggest a possible metallopeptide-based strategy for the molecular recognition of native RNA structures and insight with regard to the general features available for ligand binding site discrimination. PMID- 9724524 TI - Coulombic forces in protein-RNA interactions: binding and cleavage by ribonuclease A and variants at Lys7, Arg10, and Lys66. AB - The interactions between bovine pancreatic ribonuclease A (RNase A) and its RNA substrate extend beyond the scissile P-O5' bond. Enzymic subsites interact with the bases and phosphoryl groups of the bound substrate. Those residues interacting with the phosphoryl group comprise the P0, P1, and P2 subsites, with the scissile bond residing in the P1 subsite. Here, the function of the P0 and P2 subsites of RNase A is characterized in detail. Lys66 (P0 subsite) and Lys7 and Arg10 (P2 subsite) were replaced with alanine residues. Wild-type RNase A and the K66A, K7A/R10A, and K7A/R10A/K66A variants were evaluated as catalysts for the cleavage of poly(cytidylic acid) [poly(C)] and for their abilities to bind to single-stranded DNA, a substrate analogue. The values of kcat and Km for poly(C) cleavage were affected by altering the P0 and P2 subsites. The kcat/Km values for poly(C) cleavage by the K66A, K7A/R10A, and K7A/R10A/K66A variants were 3-fold, 60-fold, and 300-fold lower, respectively, than that of wild-type RNase A. These values indicate that the P0 and P2 subsites contribute 0.70 and 2.46 kcal/mol, respectively, to transition-state binding. Binding experiments indicate that the P0 and P2 subsites contribute 0.92 and 1.21 kcal/mol, respectively, to ground state binding. Thus, the P0 subsite makes a uniform contribution toward binding the ground state and the transition state, whereas the P2 subsite differentiates, binding more tightly to the transition state than to the ground state. In addition, nucleic acid binding to wild-type RNase A is strongly dependent on NaCl concentration, but this dependence is diminished upon alteration of the P0 or P2 subsite. The logarithm of Kd is a linear function of the logarithm of [Na+] over the range 0.018 M 2 mM Ca in the absence of NaCl. SP-A binding to vesicles does not show an absolute specificity for the phospholipid structure, but the time course of the subsequent changes does. The results suggest that SP-A contacts between phospholipid interfaces could mediate the exchange of phospholipid species (trafficking and sorting) between lung surfactant pools in the hypophase and all accessible phospholipid interfaces of the alveolar space. PMID- 9724532 TI - Inhibition of the cAMP-dependent protein kinase by synthetic A-helix peptides. AB - The catalytic subunit of the cAMP-dependent protein kinase from Dictyostelium discoideum, PkaC, displays the same properties as its mammalian counterpart, except for being about twice as large in size. Sequence comparisons indicated the presence of a conserved alpha-helix (A-helix) within the N-terminal region of PkaC which could potentially establish close contacts with the catalytic core [Veron, M., et al. (1993) Proc. Natl. Acad. Sci. U.S.A. 90, 10618-10622]. We show in this report that a synthetic peptide with the A-helix sequence inhibits PKA activity, whereas unrelated peptides display no inhibitory activity. The inhibition seems competitive with respect to the kemptide substrate rather than due to binding to a secondary site. We further show by amino acid replacements that the last lysine of the A-helix sequence is involved in this specific inhibition. A model is proposed for the possible role of the A-helix. PMID- 9724533 TI - Substrate channeling and domain-domain interactions in bifunctional thymidylate synthase-dihydrofolate reductase. AB - The thymidylate synthase (TS) and dihydrofolate reductase (DHFR) enzymes are found on a single polypeptide chain in several species of protozoa such as the parasitic Leishmania major. Earlier studies with the bifunctional TS-DHFR enzyme from L. major have suggested that this enzyme exhibits a phenomenon known as substrate channeling [Meek, T. D., et al. (1985) Biochemistry 24, 678-686]. This is a process by which a metabolite or intermediate is directly transferred from one enzyme active site to the next without being released free into solution. The crystal structure for the bifunctional TS-DHFR enzyme from L. major was recently solved, and it was shown that the TS active site was located 40 A from the DHFR active site [Knighton, D. R., et al. (1994) Nat. Struct. Biol. 1, 186-194]. On the basis of the crystal structure, a novel mechanism has been proposed for the channeling of the intermediate, dihydrofolate, from the TS active site to the DHFR active site [Knighton, D. R., et al. (1994) Nat. Struct. Biol. 1, 186-194]. They suggest that the dihydrofolate is transferred via an "electrostatic" channel on the protein surface which connects the two active sites. In this report, we describe the use of a rapid transient kinetic analysis in examining the kinetics of substrate channeling as well as domain-domain interactions in the bifunctional TS-DHFR from L. major. PMID- 9724534 TI - Kinetic reaction scheme for the dihydrofolate reductase domain of the bifunctional thymidylate synthase-dihydrofolate reductase from Leishmania major. AB - In several species of protozoa, the catalytic activities for the enzymes dihydrofolate reductase (DHFR) and thymidylate synthase (TS) reside on a single polypeptide chain constituting a bifunctional thymidylate synthase-dihydrofolate reductase enzyme. In most other species, however, these enzymes occur as monofunctional catalytic activities on separate enzymes. In this study, the kinetic reaction scheme for the dihydrofolate reductase activity from the bifunctional thymidylate synthase-dihydrofolate reductase (TS-DHFR) isolated from the parasite Leishmania major is compared to that of the monofunctional DHFR purified from Escherichia coli. Examination using pre-steady-state kinetic methods reveals interesting differences between the bifunctional and monofunctional forms of the dihydrofolate reductase enzymes. The rate-limiting step in the kinetic pathway for the monofunctional E. coli enzyme is the release of product, tetrahydrofolate. In contrast, for the L. major bifunctional enzyme, the kinetic step which limits the steady-state turnover is a conformational change associated with the release of NADP+. A complete kinetic description for the dihydrofolate reductase reaction pathway for the bifunctional enzyme is presented. PMID- 9724535 TI - Truncation of limonene synthase preprotein provides a fully active 'pseudomature' form of this monoterpene cyclase and reveals the function of the amino-terminal arginine pair. AB - The monoterpene cyclase limonene synthase transforms geranyl diphosphate to a monocyclic olefin and constitutes the simplest model for terpenoid cyclase catalysis. (-)-4S-Limonene synthase preprotein from spearmint bears a long plastidial targeting sequence. Difficulty expressing the full-length preprotein in Escherichia coli is encountered because of host codon usage, inclusion body formation, and the tight association of bacterial chaperones with the transit peptide. The purified preprotein is also kinetically impaired relative to the mixture of N-blocked native proteins produced in vivo by proteolytic processing in plastids. Therefore, the targeting sequence, that precedes a tandem pair of arginines (R58R59) which is highly conserved in the monoterpene synthases, was removed. Expression of this truncated protein, from a vector that encodes a tRNA for two rare arginine codons (pSBET), affords a soluble, tractable 'pseudomature' form of the enzyme that is catalytically more efficient than the native species. Truncation up to and including R58, or substitution of R59, yields enzymes that are incapable of converting the natural substrate geranyl diphosphate, via the enzymatically formed tertiary allylic isomer 3S-linalyl diphosphate, to (-) limonene. However, these enzymes are able to cyclize exogenously supplied 3S linalyl diphosphate to the olefinic product. This result indicates a role for the tandem arginines in the unique diphosphate migration step accompanying formation of the intermediate 3S-linalyl diphosphate and preceding the final cyclization reaction catalyzed by the monoterpene synthases. The structural basis for this coupled isomerization-cyclization reaction sequence can be inferred by homology modeling of (-)-4S-limonene synthase based on the three-dimensional structure of the sesquiterpene cyclase epi-aristolochene synthase [Starks, C. M., Back, K., Chappell, J., and Noel, J. P. (1997) Science 277, 1815-1820]. PMID- 9724536 TI - Structural and physiologic determinants of human erythrocyte sugar transport regulation by adenosine triphosphate. AB - Human erythrocyte sugar transport is mediated by the integral membrane protein GLUT1 and is regulated by cytosolic ATP [Carruthers, A., and Helgerson, A. L. (1989) Biochemistry 28, 8337-8346]. This study asks the following questions. (1) Where is the GLUT1 ATP binding site? (2) Is ATP-GLUT1 interaction sufficient for sugar transport regulation? (3) Is ATP modulation of transport subject to metabolic control? GLUT1 residues 301-364 were identified as one element of the GLUT1 ATP binding domain by peptide mapping and N-terminal sequence analysis of proteolytic fragments of azidoATP-photolabeled GLUT1. Nucleotide binding and sugar transport experiments undertaken with dimeric and tetrameric forms of GLUT1 indicate that only tetrameric GLUT1 binds and is subject to modulation by ATP. Reconstitution experiments indicate that nucleotide and tetrameric GLUT1 are sufficient for ATP modulation of sugar transport. Feedback control of GLUT1 regulation by ATP was investigated by measuring sugar uptake into erythrocyte ghosts containing or lacking ATP and glycolytic intermediates. Only AMP and ADP modulate ATP regulation of transport. Reduced cytosolic pH inhibits ATP modulation of GLUT1-mediated 3OMG uptake and increases Kd(app) for ATP interaction with GLUT1. We conclude that tetrameric but not dimeric GLUT1 is subject to direct regulation by cytosolic ATP and that this regulation is antagonized by intracellular AMP and acidification. PMID- 9724537 TI - Channel-lining residues in the M3 membrane-spanning segment of the cystic fibrosis transmembrane conductance regulator. AB - The cystic fibrosis transmembrane conductance regulator (CFTR) forms a chloride selective channel. Residues from the 12 putative membrane-spanning segments form at least part of the channel lining. We need to identify the channel-lining residues in order to understand the structural basis for the channel's functional properties. Using the substituted-cysteine-accessibility method we mutated to cysteine, one at a time, 24 consecutive residues (Asp192-Ile215) in the M3 membrane-spanning segment. Cysteines substituted for His199, Phe200, Trp202, Ile203, Pro205, Gln207, Leu211, and Leu214 reacted with charged, sulfhydryl specific reagents that are derivatives of methanethiosulfonate (MTS). We infer that these residues are on the water-accessible surface of the protein and probably form a portion of the channel lining. When plotted on an alpha-helical wheel the exposed residues from Gln207 to Leu214 lie within an arc of 60 degrees; the exposed residues in the cytoplasmic half (His199-Ile203) lie within an arc of 160 degrees. We infer that the secondary structures of the extracellular and cytoplasmic halves of M3 are alpha-helical and that Pro205, in the middle of the M3 segment, may bend the M3 segment, moving the cytoplasmic end of the segment in toward the central axis of the channel. The bend in the M3 segment may help to narrow the channel lumen near the cytoplasmic end. In addition, unlike full length CFTR, the current induced by the deletion construct, Delta259, is inhibited by the MTS reagents, implying that the channel structure of Delta259 is different than the channel structure of wild-type CFTR. PMID- 9724538 TI - Modulation of the binding of signal peptides to lipid bilayers by dipoles near the hydrocarbon-water interface. AB - Interactions between signal (leader) sequences and membranes are critical to protein insertion and translocation across membranes. In this paper, circular dichroism, tryptophan fluorescence, electrophoretic mobility, dipole potential, and binding measurements were used to study the interaction of the signal sequence of the Escherichia coli LamB protein with various lipid bilayers. By modifying specific chemicophysical properties of both the signal sequence and bilayer, we analyzed some of the key factors underlying peptide-lipid interactions. We synthesized three analogues of the LamB signal peptide differing in their net charge (-2 to +4) and studied their binding to bilayers containing combinations of neutral lipids [egg phosphatidylcholine (EPC), sphingomyelin, cholesterol, ketocholesterol, and nitroxide-containing phospholipid] and a charged lipid (phosphatidylserine). All three peptides bound to EPC bilayers and underwent a random coil to alpha-helix transition upon binding. Microelectrophoresis experiments revealed that both the N and C termini were near the outer surface of the bilayer, suggesting that the peptides adopted a "hammock" configuration with both termini exposed to the aqueous phase and the core of the alpha-helix located near the hydrocarbon-water interface. The binding of these LamB peptides was not markedly dependent on the bilayer area per molecule, compressibility modulus, or dipole potential, but did depend on the charge of the peptide and bilayer interfacial region. Moreover, the binding of LamB peptides was essentially eliminated in bilayers composed of phospholipids with a nitroxide moiety at the 7 position in one of their acyl chains or in EPC bilayers containing equimolar ketocholestanol. We propose that the incorporation of nitroxide or ketone groups into the hydrocarbon region near the lipid headgroup increases the effective width of the hydrophilic interfacial region and prevents some of the hydrophobic amino acids in the alpha-helix from reaching the nonpolar hydrocarbon core, thereby diminishing the free energy of partitioning and inhibiting peptide binding. These results point to an important role for interfacial dipoles in peptide-lipid interactions. PMID- 9724539 TI - A recombinant monocysteine mutant (Ser to Cys-155) of fast skeletal troponin T: identification by cross-linking of a domain involved in a physiologically relevant interaction with troponins C and I. AB - Troponin T (TnT), a subunit of the heterotrimeric troponin (Tn) complex, is essential for the Ca2+ regulation of vertebrate striated muscle contraction both in vivo and in vitro. With the exception of bovine cardiac TnT, all known vertebrate TnT isoforms lack a thiol group, a property which makes the wild-type proteins unsuitable as cross-linking substrate. We generated a mutant human fast skeletal TnT in which Ser155 was changed to Cys (TnT-Cys155). Mutation of this residue in TnT as well as in vitro expression in Escherichia coli and purification of the recombinant mutant protein did not affect its biological properties in terms of in vitro binding to troponin I (TnI), troponin C (TnC), actin-tropomyosin (actin-Tm), and actomyosin ATPase activity. TnT-Cys155 was labeled with 4-maleimidobenzophenone (BP-TnT155) and photo-cross-linked to TnI, TnC, Tm, and all of the thin filament proteins. BP-TnT155 did not cross-link to Tm and showed weak Ca2+/Mg2+-independent cross-linking with TnI in the binary complex and in the presence of all thin filament protein components. BP-TnT155 showed Ca2+/Mg2+-dependent cross-linking with TnC in the binary and ternary complexes and Ca2+-favored cross-linking with TnI in the ternary complex. Thus, residue 155 of TnT is within 10 A (the length of cross-linker) of TnC in the presence or absence of Ca2+ and comes within 10 A of both TnI and TnC in the presence of Ca2+. TnT residue 155 is in close proximity to or may even partly encompass the Tm binding site. These results suggest that TnT, in association with TnI, may participate in the "information transfer" mediated by the Ca2+ binding signal from TnC to Tm and the region around TnT residue 155 probably acts as a linker between troponin and actin-Tm in this signal transmission process. Our results also suggest that TnT contains at least one Ca2+/Mg2+-dependent TnC binding region located between its Tm and TnI binding regions. A recombinant truncated fragment of TnI, TnI96-181, containing amino acid residues 96-181 and labeled with BP at Cys-133, failed to cross-link with TnT, indicating that the region around Cys-133 of TnI is not involved in binary interaction with TnT. PMID- 9724540 TI - Translocase-bound SecA is largely shielded from the phospholipid acyl chains. AB - Protein translocation in Escherichia coli is mediated by the SecA ATPase bound to the SecYEG membrane protein complex. SecA translocation ATPase activity as well as protein translocation is dependent on the presence of negatively charged lipids. By using a phospholipid with an acyl chain linked photoactivatable group, the lipid accessibility of SecA bound at the translocase was explored. SecA bound to lipid vesicles containing negatively charged lipids was found to be readily accessible for labeling by the photoactivatable phospholipid. The presence of an excess amount of SecYEG complex resulted in a remarkable reduction in the amount of lipid-accessible SecA irrespective of the nucleotide-bound form of SecA. These data demonstrate that the SecYEG-bound SecA is largely shielded from the phospholipid acyl chains and suggest the presence of two distinct pools of membrane-bound SecA that differ in the degree of lipid association. PMID- 9724541 TI - TNP-ATP and TNP-ADP as probes of the nucleotide binding site of CheA, the histidine protein kinase in the chemotaxis signal transduction pathway of Escherichia coli. AB - The interaction of CheA with ATP has important consequences in the chemotaxis signal transduction pathway of Escherichia coli. This interaction results in autophosphorylation of CheA, a histidine protein kinase. Autophosphorylation of CheA sets in motion a chain of biochemical events that enables the chemotaxis receptor proteins to communicate with the flagellar motors. As a result of this communication, CheA allows the receptors to control the cell swimming pattern in response to gradients of attractant and repellent chemicals. To probe CheA interactions with ATP, we investigated the interaction of CheA with the fluorescent nucleotide analogues TNP-ATP [2'(3')-O-(2,4,6 trinitrophenyl)adenosine 5'-triphosphate] and TNP-ADP. Spectroscopic studies indicated that CheA bound TNP-ATP and TNP-ADP with high affinity (micromolar Kd values) and caused a marked enhancement of the fluorescence of the TNP moiety of these modified nucleotides. Analysis of titration experiments indicated a binding stoichiometry of two molecules of TNP-ATP (TNP-ADP) per CheA dimer and suggested that the two binding sites on the CheA dimer operate independently. Binding of TNP-ATP to CheA was inhibited by ATP, and analysis of this inhibition indicated that the CheA dimer binds 2 molecules of ATP. Competition experiments also indicated that CheA binds TNP-ATP considerably more tightly than it binds unmodified ATP. Binding of TNP-ADP to CheA was inhibited by ADP in a similar manner. TNP-ATP was not a substrate for CheA and served as a potent inhibitor of CheA autophosphorylation (Ki < 1 microM). The glycine-rich regions (G1 and G2) of CheA and other histidine protein kinases have been presumed to play important roles in ATP binding and/or catalysis of CheA autophosphorylation, although few experimental tests of these functional assignments have been made. Here, we demonstrate that a CheA mutant protein with Gly-->Ala substitutions in G1 and G2 has a markedly reduced affinity for ATP and ADP, as measured by Hummel-Dreyer chromatography. This mutant protein also bound TNP-ATP and TNP-ADP very poorly and had no detectable autokinase activity. Surprisingly, a distinct single-site substitution in G2 (Gly470-->Lys) had no observable effect on the affinity of CheA for ATP and ADP, despite the fact that it rendered CheA completely inactive as an autokinase. This mutant protein also bound TNP-ATP and TNP-ADP with affinities and stoichiometries that were indistinguishable from those observed with wild-type CheA. These results provide some preliminary insight into the possible functional roles of G1 and G2, and they suggest that TNP-nucleotides are useful tools for exploring the effects of additional mutations on the active site of CheA. PMID- 9724542 TI - Heterogeneous processing of a G protein gamma subunit at a site critical for protein and membrane interactions. AB - The G protein gamma5 subunit is selectively associated with specific G protein alpha subunits [Wilcox, M. D., et al. (1995) J. Biol. Chem. 270, 4189] and is localized preferentially in focal adhesion plaques [Hansen, C. A., et al. (1996) J. Cell Biol. 126, 811]. What determines the differential association of G proteins and their subunits with specific cellular structures or compartments is not clear, but one factor could be variation in the pattern of processing of the proteins. To study gamma5 subunit diversity and modifications, G protein subunits were fractionated on an HPLC phenyl column and analyzed with a gamma5-specific antiserum. The gamma5 eluted from the column as two peaks of immunoreactivity. Analysis by matrix-assisted laser desorption ionization (MALDI) mass spectrometry and electrospray ionization tandem mass spectrometry revealed that the first immunoreactive peak corresponded to the predicted gamma5 isoform (N-terminally acetylated after removal of methionine, C-terminally geranylgeranylated and carboxymethylated with removal of the last three amino acids), while the second peak of immunoreactivity contained a gamma5 isoform isoprenylated at the C terminus but retaining its three terminal amino acids. This alternatively processed protein is the predominant gamma5 subunit isoform associated with Go and Gi proteins purified from bovine brain. These results describe a new C terminal processing pattern for G protein gamma subunits and establish the principle that G protein gamma subunits can be heterogeneously modified at their C-termini. This is a site on the gamma subunit critical for membrane and protein protein interactions of G proteins. These results open the possibility that one determinant of the localization of G proteins in cells could be the pattern of processing of their gamma subunit constituents. PMID- 9724543 TI - Cu,Zn superoxide dismutase from Photobacterium leiognathi is an hyperefficient enzyme. AB - The catalytic rate constant of recombinant Photobacterium leiognathi Cu,Zn superoxide dismutase has been determined as a function of pH by pulse radiolysis. At pH 7 and low ionic strength (I = 0.02 M) the catalytic rate constant is 8.5 x 10(9) M-1 s-1, more than two times the value found for all the native eukaryotic Cu,Zn superoxide dismutases investigated to date. Similarly, Brownian dynamics simulations indicate an enzyme-substrate association rate more than two times higher than that found for bovine Cu,Zn superoxide dismutase. Titration of the paramagnetic contribution to the water proton relaxation rate of the P. leiognathi with increasing concentration of halide ions with different radii indicates that the proteic channel delimiting the active site is wider than 4.4 A. This is at variance with that found on the eukariotic enzymes, and provides a rationale for the high catalytic rate of the bacterial enzyme. Evidence for solvent exposure of the active site different from that observed in the eukaryotic enzyme is suggested from the pH dependence of the water proton relaxation rate and of the EPR spectrum line shape, which indicate the occurrence of a prototropic equilibrium at pH 9.1 and 9.0, respectively. The pH dependence of the P. leiognathi catalytic rate has a trend different from that observed in the bovine enzyme, indicating that groups differently exposed to the solvent are involved in the modulation of the enzyme-substrate encounter. PMID- 9724544 TI - Membrane-bound cytochrome cz couples quinol oxidoreductase to the P840 reaction center complex in isolated membranes of the green sulfur bacterium Chlorobium tepidum. AB - The reaction of quinol oxidoreductase and membrane-bound c-type cytochromes was studied in chlorosome-depleted membranes isolated from Chlorobium tepidum. Rapid oxidations of c-type cytochromes were detected after flash excitation. Their re reductions occurred in parallel with the reduction of cytochrome b, especially in the presence of antimycin A, whereas reductions of both cytochromes c and b were suppressed by added stigmatellin. These results indicate the tight coupling between the photosynthetic reaction center and quinol oxidoreductase. Turnovers of two types of cytochromes c were detected. One was assigned to the monoheme type cytochrome c (designated cytochrome cz), which is known to be tightly bound to the reaction center complex. The other was a new c-type cytochrome, cytochrome c-556, which functions the same as cytochrome c1. The steps of electron-transfer scheme, menaquinol --> Rieske FeS center --> cytochrom c-556 --> cytochrome cz - > P840, are estimated to have reaction times of 20 ms and 560, 150, and 40 microseconds, respectively. We conclude that quinol oxidoreductase and the reaction center complex in Chlorobium tepidum are linked by two distinct membrane bound cytochromes, cz and c-556, with no involvement of water-soluble cytochromes. PMID- 9724545 TI - Functional implications of the proximal hydrogen-bonding network in myoglobin: a resonance Raman and kinetic study of Leu89, Ser92, His97, and F-helix swap mutants. AB - Resonance Raman spectra have been obtained for both the equilibrium deoxy derivative and the 10 ns photoproduct of the CO derivative of several mutants of sperm whale myoglobin. The particular mutations on the F-helix were chosen to expose the role of the proximal hydrogen-bonding network in maintaining the position of the heme, the proximal histidine, and the heme-7-propionate. In each mutant, one or more hydrogen bonds are altered or eliminated. A careful comparison of the spectra from the equilibrium and transient five coordinate species indicates that the tertiary relaxation after photodissociation is nearly complete within 10 ns, as is the case in the WT protein. The iron-proximal histidine stretching mode (nu(Fe-His)) and several low-frequency propionate sensitive modes in the Raman spectra reveal the impact of specific disruptions in the hydrogen-bonding network on the heme pocket geometry. Two categories of perturbation are observed with respect to nu(Fe-His): (1) a shift in the peak frequency without a change in line shape and (2) changes in the overall line shape which may or may not be accompanied by a frequency shift. The alterations in the nu(Fe-His) band are interpreted as arising from conformational heterogeneity and local geometrical changes within the pocket, including movement of the heme group, and are discussed in terms of changes in the population distribution as revealed via a curve-fitting analysis. None of the frequency shifts in the nu(Fe-His) band are as large as that reported for the His93Gly(imidazole) mutant, suggesting that the covalent linkage between the heme and His93 plays a crucial role in maintaining the geometry of the proximal pocket. Molecular modeling indicates that the nu(Fe-His) frequency shifts observed in the present study originate from changes in the His93 imidazole ring azimuthal angle. The systematic variations in the interactions of the heme-7 propionate in the mutants have exposed several properties of the propionate sensitive Raman bands. The frequencies of nu9 (the 240 cm-1 shoulder on the nu(Fe His) band) and delta(cbetacccd) at approximately 370 cm-1 appear to be correlated. A decrease in hydrogen-bond strength to this propionate in response to changes in stereochemistry or degree of disorder is associated with a decrease in the frequency of both nu9 and delta(cbetacccd). The mutations that cause a weakening of the hydrogen bonding to the heme-7-propionate also result in changes in nu(Fe-His) which are interpreted as evidence that this propionate participates in the anchoring of the heme within the heme pocket. Changes in gamma7 at approximately 300 cm-1, gamma6 at approximately 335 cm-1, and nu8 at approximately 342 cm-1 are discussed in terms of pocket disorder. A titration from pH 5.1 to 7.4 suggests that His97 is protonated in the WT protein by pH 5.1. Geminate-rebinding studies on these mutants indicate that disruption of the hydrogen-bonding network has only modest effects on ligand-binding kinetics, suggesting that the role of the hydrogen-bonding network may be one of maintaining heme pocket stability rather than of specific protein function. PMID- 9724546 TI - Probing the backbone dynamics of oxidized and reduced rat microsomal cytochrome b5 via 15N rotating frame NMR relaxation measurements: biological implications. AB - Rotating frame 15N relaxation NMR experiments have been performed to study the local mobility of the oxidized and reduced forms of rat microsomal cytochrome b5, in the microsecond to millisecond time range. Measurements of rotating frame relaxation rates (R1rho) were performed as a function of the effective magnetic field amplitude by using off-resonance radio frequency irradiation. Detailed analysis of the two data sets resulted in the identification of slow motions along the backbone nitrogens for both oxidation states of the protein. The local mobility of reduced and oxidized cytochrome b5 turned out to be significantly different; 28 backbone nitrogens of the oxidized form were shown to participate in a conformational exchange process, while this number dropped to 12 in the reduced form. The correlation time, tauex, for the exchange processes could be determined for 21 and 9 backbone nitrogens for oxidized and reduced cytochrome b5, respectively, with their values ranging between 70 and 280 microseconds. The direct experimental evidence provided in this study for the larger mobility of the oxidized form of the protein is consistent with the different backbone NH solvent exchangeability recently documented for the two oxidation states [Arnesano, F., et al. (1998) Biochemistry 37, 173-184]. Our experimental observations may have significant biological implications. The differential local mobility between the two oxidation states is proposed to be an important factor controlling the molecular recognition processes in which cytochrome b5 is involved. PMID- 9724547 TI - Hybridization of peptide nucleic acid. AB - The thermodynamics of hybridization and the conformations of decameric mixed purine-pyrimidine sequence PNA/PNA, PNA/DNA, and DNA/DNA duplexes have been studied using fluorescence energy transfer (FET), absorption hypochromicity (ABS), isothermal titration calorimetry (ITC), and circular dichroism (CD) techniques. The interchromophoric distances determined in the FET experiments on fluorescein- and rhodamine-labeled duplexes indicate that the solution structures of the duplexes are extended helices in agreement with available NMR (PNA/DNA) and crystal X-ray data (PNA/PNA). The melting thermodynamics of the duplexes was studied with both FET and ABS. The thermodynamic parameters obtained with ABS are in good agreement with the parameters from calorimetric measurements while FET detection of duplex melting gives in most cases more favorable free energies of hybridization. This discrepancy between FET and ABS detection is ascribed to the conjugated dyes which affect the stability of the duplexes substantially. Especially, the dianionic fluorescein attached via a flexible linker either to PNA or to DNA seems to be involved in an attractive interaction with the opposite dicationic lysine when hybridized to a PNA strand. This interaction leads to an increased thermal stability as manifested as a 3-4 degreesC increase of the melting temperature. For the PNA/DNA duplex where fluorescein is attached to the PNA strand, a large destabilization (DeltaTm = -12 degreesC) occurs relative to the unlabeled duplex, probably originating from electrostatic repulsion between the fluorescein and the negatively charged DNA backbone. In the case of the PNA/PNA duplex, the sense of helicity of the duplex is reversed upon conjugation of fluorescein via a flexible linker arm, but not when the fluorescein is attached without a linker to the PNA. PMID- 9724548 TI - Differences between DNA base pair stacking energies are conserved over a wide range of ionic conditions. AB - Base pair stacking free energy parameters in a low ionic strength solvent were determined from an analysis of DNA fragments using temperature gradient gel electrophoresis (TGGE). Transition midpoint temperatures (Tu) were determined for the first melting domain (52 +/- 4 bp) of 16, 339 bp DNAs that differed from each other by single base pair substitutions. The data were combined with previously obtained Tu data from 17 similar DNAs that had single base pair changes at different sites [Ke, S. H., and Wartell, R. M. (1995) Biochemistry 34, 4593 4599]. The Tu values were used to evaluate free energy differences (deltaDeltaG) between 31 pairs of DNAs. Linear equations relating the deltaDeltaG values to changes in base pair stacking were analyzed by singular value decomposition (SVD) to determine the 10 nearest neighbor free energy parameters. The order of stability of the parameters, TA < AT < AA < AG < GT approximately TC approximately TG < CC < GC approximately CG, was essentially the same as the hierarchy determined in 1 M Na+ [Allawi, H. T., and SantaLucia, J., Jr. (1997) Biochemistry 36, 10581-10594]. The experimental free energy differences were in good agreement with predictions made using nearest-neighbor parameters determined from several previous studies conducted in medium or high salt concentrations. Conversely the parameters determined in the current study produced good predictions of free energy differences previously determined from 59 DNA oligomers in 1 M Na+. The results indicate that differences between base pair stacking energies are conserved across a wide range of ionic conditions, and in both oligomer and polymer DNA contexts. PMID- 9724549 TI - Conformational state of ovalbumin at acidic pH as evaluated by a novel approach utilizing intrachain sulfhydryl-mixed disulfide exchange reactions. AB - Ovalbumin contains four cysteine sulfhydryls (Cys11, Cys30, Cys367, and Cys382) and one cystine disulfide (Cys73-Cys120). A highly reactive aromatic disulfide, 2,2'-dipyridyl disulfide, reacts specifically with Cys367 of ovalbumin at pH 2.2 generating a mixed disulfide protein derivative [Tatsumi, E., and Hirose, M. (1997) J. Biochem. 122, 300-308]. The mode of conformational fluctuation in ovalbumin was investigated at pH 2.2 using the mixed disulfide derivatives of the cystine-intact and cystine-reduced protein forms. In the presence of a high concentration of urea, both the mixed disulfide derivatives underwent rapid cysteine sulfhydryl/mixed disulfide exchanges, thereby releasing the quantitative amount of 2-thiopyridone. A peptide mapping analysis for disulfide-forming cysteines revealed that this release was mostly accounted for by the nucleophile attack on the Cys367-mixed disulfide by the nearest cysteine residue in the primary structure, Cys382. At the acidic pH, the exchange reaction was practically restricted to the cysteine sulfhydryl/mixed disulfide exchanges; no other exchange reaction, such as the cysteine sulfhydryl/cystine disulfide exchange reaction, was detected. In the absence of urea, the cystine-reduced form, but not the cystine-intact form, underwent significant sulfhydryl/mixed disulfide exchange reactions at a physiological temperature, as determined by the release of 2-thiopyridone. A kinetic analysis for the generation of disulfide forming cysteines with Cys367 at 37 degreesC revealed that the rate for the intrachain exchange reaction was quite different for the five cysteine sulfhydryls. The effective concentrations of the five cysteine sulfhydryls relative to the Cys367-mixed disulfide were determined by using three related model reactions: the obtained values were 11.4, 4.6, 15.2, 5.9, and 8.9 microM for Cys11, Cys30, Cys73, Cys120, and Cys382, respectively. Implications of the effective concentrations for the conformational state of acidic ovalbumin are discussed. PMID- 9724550 TI - Arginine-42 and threonine-45 are required for FAD incorporation and catalytic activity in human monoamine oxidase B. AB - Monoamine oxidase B (MAO B) is an integral protein of the outer mitochondrial membrane that is involved in the deamination of vasoactive and neuroactive amines. The oxidation of these amine substrates requires the cofactor FAD, which is covalently bound to Cys-397 of human MAO B. Previously, Glu-34 and Tyr-44 of MAO B have been identified as residues which engage in noncovalent interactions with FAD that are required for subsequent covalent FAD binding and generation of catalytic activity. In this study, we have identified two additional residues, Arg-42 and Thr-45, which form noncovalent contacts with FAD that are prerequisite steps to the covalent attachment of FAD. Arg-42 and Thr-45, along with Tyr-44, comprise part of a highly conserved flavin binding sequence, RXY(T,S), that is found in other flavoproteins, several of which have well-defined X-ray crystal structures. We tested the roles of Arg-42 and Thr-45 in MAO B by constructing mutant MAO B cDNAs which encode amino acid substitutions at these residues and expressed the variant proteins in COS-7 cells. Substitution of Arg-42 or Thr-45 with alanine resulted in complete loss of MAO B activity and FAD incorporation. However, conservative substitutions of Arg-42 with lysine or Thr-45 with serine resulted in MAO B variants that retain both partial activity and partial FAD incorporation. These results indicate that Arg-42 and Thr-45 form critical noncovalent interactions with FAD that are required for the subsequent activation of MAO B by covalent coupling of FAD. PMID- 9724551 TI - The role of P-glycoprotein and canalicular multispecific organic anion transporter in the hepatobiliary excretion of drugs. PMID- 9724552 TI - Caco-2 cell permeability of a new (hydroxybenzyl)ethylenediamine oral iron chelator: correlation with physicochemical properties and oral activity. AB - This study describes the transport of CGP 75254A, a novel oral iron chelator, across Caco-2 cells in an attempt to model intestinal epithelial cell permeability in man. CGP 75254A was dosed to the apical side of Caco-2 cell monolayers, together with [14C]mannitol as an internal permeability standard. The apparent permeability (Papp) was calculated from the cumulative appearance of drug in the basolateral fluid with time. The [14C]mannitol Papp indicated that the Caco-2 monolayers remained intact and that the iron chelator was not toxic to the cells. Permeabilities of CGP 75254A were compared with the Caco-2 permeabilities of compounds of known absorption in man. The results predict that absorption of CGP 75254A is likely to be virtually complete at pH values between 5.5 and 7.0. However, at pH 8.0 permeability is predicted as negligible. Cell permeability data are in full accordance with key physicochemical properties of CGP 75254A and suggest that the drug is passively absorbed. The results, which suggest likely quantitative absorption in vivo, are supported by preliminary pharmacological experiments in marmosets. PMID- 9724553 TI - Stability of lipid/DNA complexes during agitation and freeze-thawing. AB - It is well established that cationic liposomes facilitate the delivery of DNA and offer substantial advantages over viral-based delivery systems. However, these synthetic vectors readily aggregate in liquid formulations which in clinical trials requires preparation of lipid/DNA complexes at the bedside immediately before injection. This temporal requirement could be eliminated if complexes were formulated as stable preparations that could be shipped, stored, and administered as needed. To this end, our study investigates the stability of lipid/DNA complexes during physical stresses that might be encountered during shipping and storage, i.e., agitation and freeze-thawing. Our data show that agitation significantly reduces transfection rates in complexes prepared with three different commercially available lipid formulations. Additional experiments indicate that slow freezing is more damaging than rapid freezing, and that sucrose is able to preserve transfection and complex size during freeze-thawing. These results are consistent with previous reports and demonstrate that frozen formulations may be suitable for maintaining transfection rates of lipid/DNA complexes. Under certain conditions, we observe a reproducible 3-fold increase in transfection after freeze-thawing that is prevented by high concentrations of sucrose. Together, these data suggest that physical stresses can alter structural characteristics of lipid/DNA complexes that can markedly affect rates of DNA delivery. PMID- 9724554 TI - Physical characterization of nedocromil sodium hydrates. AB - Nedocromil sodium, which is used in the treatment of reversible obstructive airway diseases, such as asthma, is found to exist in the following hydrate phases: the heptahemihydrate, the trihydrate, a monohydrate, and an amorphous phase which contains variable amounts of water (1.5-3.0 mol). An anhydrate phase is formed from the trihydrate at zero humidity at >/= 150 degrees C, but is rapidly hydrated under ambient conditions. The physical and thermodynamic properties of the four hydrate phases were characterized using differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), powder X-ray diffraction (PXRD) at ambient and elevated temperatures, hot-stage microscopy (HSM), solid phase interconversion at various relative humidities (RH), intrinsic dissolution rate (IDR), equilibrium solubility measurements, and critical RH measurements. Below 100 degrees C in open pan TGA, the heptahemihydrate and the amorphous forms lose virtually all their water, the monohydrate loses negligible amounts of water, whereas the trihydrate loses the first two moles of water. From 130 degrees C to 200 degrees C in open pan TGA the trihydrate and the monohydrate lose their last mole of water to form the anhydrate. In crimped pan DSC, the thermal events observed are analogous to those observed in open pan TGA, but the temperatures are increased by about 75 degreesC for all except the heptahemihydrate, for which the thermal events are more complex. When the heptahemihydrate is heated in a crimped pan, a melting endotherm is observed at about 75 degrees C followed by three dehydration endotherms. For the crystalline hydrate phases at 22 degrees C, the ranges of stability are as follows: the monohydrate from 0 to 6.4% RH; the trihydrate from 6.4 to 79.5% RH; the heptahemihydrate above 80% RH. A microbalance study showed that the heptahemihydrate is kinetically stable over the range 11 to 79.5% RH. The IDR in water at 25 degrees C under constant hydrodynamic conditions decreases in the rank order: monohydrate > trihydrate > heptahemihydrate, corresponding to the rank order of free energy with respect to the aqueous solution. The equilibrium aqueous solubility of the heptahemihydrate at 25.0 +/- 0.2 degrees C is 0.956 +/- 0.010 M. PMID- 9724555 TI - Effects of Tween 80 and sucrose on acute short-term stability and long-term storage at -20 degrees C of a recombinant hemoglobin. AB - The addition of low levels of surfactant polyoxyethylene 20 sorbitan monooleate, Tween 80, to recombinant hemoglobin in phosphate-buffered saline minimized the level of protein aggregation during acute freeze-thaw studies. Addition of sucrose alone to the phosphate-buffered saline formulation, up to 0.5 M, provided minimal protection against freeze-thaw induced aggregation. In contrast to the acute stability studies, long-term storage at -20 degrees C induced aggregation and methemoglobin formation in those formulations containing only Tween 80 in phosphate-buffered saline. Addition of sucrose between 0.1 and 0.5 M to the formulation prevented formation of aggregates and severely arrested methemoglobin formation during the long-term -20 degrees C storage. Specific binding of Tween 80 to the hemoglobin was not observed using 16-doxyl stearic acid partitioning techniques with electron paramagnetic resonance. Minor structural changes to the protein secondary structure during freezing in the absence and presence of Tween 80 were observed with Fourier transform infrared spectroscopy. The alterations were partially prevented by addition of the sucrose. It is likely that the Tween 80 severely reduced protein aggregation during the acute stability studies by preventing the hemoglobin from reaching the air-liquid interface or the liquid surface interfaces. The reduction in methemoglobin formation and aggregation observed during long-term storage can be accounted for on the premise that the sucrose reduced localized unfolding of the protein in a manner similar to the preferential exclusion theory (Arakawa, T.; and Timasheff, S. N. 1982, Biochemistry 1982, 21, 6536-6544). These studies demonstrate that acute formulation screening studies, albeit useful, may not necessarily predict protein stability during long-term storage. PMID- 9724556 TI - Aggregation of recombinant human interferon gamma: kinetics and structural transitions. AB - Protein aggregation is a complex phenomenon that can occur in vitro and in vivo, usually resulting in the loss of the protein's biological activity. While many aggregation studies focus on a mechanism due to a specific stress, this study focuses on the general nature of aggregation. Recombinant human interferon-gamma (rhIFN-gamma) provides an ideal model for studying protein aggregation, as it has a tendency to aggregate under mild denaturing stresses (low denaturant concentration, temperature below the Tm, and below pH 5). All of the aggregates induced by these stresses have a similar structure (high in intermolecular beta sheet content and a large loss of alpha-helix) as determined by infrared and circular dichroism spectroscopy. Thermally induced and denaturant-induced aggregation processes follow first-order kinetics under the conditions of this study. Spectroscopic and kinetic data suggest that rhIFN-gamma aggregates through an intermediate form possessing a large amount of residual secondary structure. In contrast to the aggregates formed under denaturing stresses, the salted-out protein has a remarkably nativelike secondary structure. PMID- 9724557 TI - Pharmaceutical properties of related calanolide compounds with activity against human immunodeficiency virus. AB - The present studies were undertaken to compare the relative pharmacokinetic parameters and bioavailability of two chemically related natural products which are nonnucleoside inhibitors of reverse transcriptase. Both (+)-calanolide A (Cal A; NSC 675451) and (+)-dihydrocalanolide A (DHCal A; NSC 678323) are currently under development for the treatment of HIV infections. HPLC-based analytical assays were developed for both compounds using modifications of a previously published procedure. The assays were used to compare the intravenous pharmacokinetics of the dihydro analogue relative to the parent compound, Cal A, and to determine the relative oral bioavailability of each drug in CD2F1 mice. Although the pharmacokinetic parameters of each drug were similar (Cal A, 25 mg/kg: AUC: 9.4 [microg/mL]. hr, t1/2beta: 0.25 h,, t1/2gamma: 1.8 h, clearance: 2.7 L/h/kg versus DHCal A, 25 mg/kg: AUC: 6.9 [microg/mL].hr, t1/2beta: 0.22 h,, t1/2gamma: 2.3 h, clearance: 3.6 L/h/kg), the oral bioavailability of DHCal A (F = 46. 8%) was markedly better than that obtained for Cal A (F = 13.2%). The relative ability of Cal A and DHCal A to change to their inactive epimer forms, (+)-calanolide B and (+)-dihydrocalanolide B, respectively, was also determined. While conversion of active to inactive forms of the drugs was noted to occur in vitro especially under acidic conditions, no epimer forms of either compound were noted in plasma of mice after administration of either CalA or DHCal A. Considered together with preliminary toxicology findings, the pharmacokinetic data obtained in the present series of experiments suggest that selection of the dihydro derivative of (+)-calanolide A may be a reasonable choice for further preclinical development and possible Phase I clinical evaluation. PMID- 9724558 TI - Absorption behavior of orally administered drugs in rats treated with propantheline. AB - The effect of gastrointestinal (GI) transit rate on the absorption behavior of orally administered drugs was investigated using rats pretreated with propantheline. The propantheline-treatment reduced the transit rate in all segments to approximately 50%. The absorption behavior was examined for three model drugs with different absorption characteristics: theophylline as a highly absorbable drug without the first-pass elimination, ampicillin as a poorly absorbable one, and cephalexin as a highly absorbable one via carrier-mediated transport system. In the GI transit-retarded state, the Tmax of the plasma concentration-time curve was delayed in all the three drugs. However, the extent of bioavailability was not changed in theophylline and cephalexin. On the other hand, the extent of bioavailability of ampicillin was increased in rats pretreated with propantheline. This might be caused by the increased residence time in the absorption site, i.e., small intestine. These results were generally predicted by use of the convolution method based on the GI-Transit-Absorption Model, which was developed in our previous study, using the GI transit rate parameters in rats pretreated with propantheline. The analysis using this model could clarify that the substantial absorption site of cephalexin moved to the upper region of the small intestine by the reduction of the GI transit rate. PMID- 9724559 TI - Simulating lipophilicity of organic molecules with a back-propagation neural network. AB - From a training set of 7200 chemicals, a back-propagation neural network (BNN) model was developed for calculating the 1-octanol/water partition coefficient (log P) of molecules containing nitrogen, oxygen, halogen, phosphorus, and/or sulfur atoms. Chemicals were described by means of autocorrelation vectors encoding hydrophobicity, molar refractivity, H-bonding acceptor ability, and H bonding donor ability. A 35/32/1 composite network composed of four configurations was selected as the final model (root-mean-square error (RMS) = 0.37, r = 0.97) because it provided the best simulation results (RMS = 0.39, r = 0.98) on an external testing set of 519 molecules. This final model compared favorably with a recently published BNN model using variables (atoms and bonds) derived from connection matrices. PMID- 9724560 TI - Isomerization of ceftibuten in aqueous solution. AB - The isomerization reactions of ceftibuten and ceftibuten-related compounds in aqueous solution were investigated to estimate the substitution effect on the isomerization reaction and identify the three proximal dissociation constants of ceftibuten kinetically from the pH-rate profiles. The isomerization reaction of ceftibuten-related compounds was influenced by the substituents near the double bond at the C7-side chain, and the electron-withdrawing substituent was found to increase the isomerization rate. Ceftibuten isomerized at the C7-side chain, and the isomerization rate was influenced by the dissociation of the C7-side chain carboxylic acid and aminothiazole in the acidic pH region. The dissociation constants of ceftibuten were assigned by comparing the isomerization rates of ceftibuten with its related compounds at various pH conditions, and the pKas 2.3, 3.2, and 4.5 were attributed to the 4-carboxylic acid, 7-carboxylic acid, and 7 aminothiazole, respectively. PMID- 9724561 TI - Stereoselective pharmacokinetics of chlorpheniramine and the effect of ranitidine. AB - This single-dose, randomized, crossover study was carried out to investigate the potential effect of ranitidine on the pharmacokinetics of chlorpheniramine. The study also afforded an opportunity to add to the limited data currently available on the stereoselective pharmacokinetics of chlorpheniramine. Healthy subjects received a single oral 4 mg dose of racemic chlorpheniramine on two separate occasions: alone, and on day 6 of dosing with ranitidine 75 mg b.i.d. for 8 days. Serum concentrations and urinary recovery of (S)-(+)- and (R)-(-) chlorpheniramine were unaffected by administration of ranitidine, indicating no pharmacokinetic drug-drug interaction. The observed chlorpheniramine pharmacokinetic data were consistent with previous data and indicated approximately 2.5-fold higher serum concentrations of the (S)-(+) enantiomer. Previously reported high variability in chlorpheniramine pharmacokinetics was greatly reduced by well-controlled food and fluid intake. PMID- 9724562 TI - A theoretical study of the interaction of anhydrotetracycline with Al(III). AB - In this article the complexation of anhydrotetracycline (AHTC), the major toxic decomposition product of the antibiotic tetracycline, with Al(III) has been investigated using the AM1 semiempirical and ab initio Hartree-Fock levels of theory. Different modes of complexation have been considered with the structure of tetra- and pentacoordinated complexes being fully optimized. In the gas phase, processes ii and iii, which lead to the complexes with stoichiometry MHL2+, are favored. Structure II ([AlLH2(OH)(H2O)]2+) has the metal coordinated to the O11 and O12 groups and the O3 group protonated and is the global minimum on the potential energy surface for the interaction. In water solution, the Al(III) is predicted to form predominantly a tetracoordinated complex at the Oam and O3 site (V) of the AHTC with the stoichiometry MH2L3+ (process i). The experimental proposal is the complexed form with the metal ion coordinated to the O11-O12 moiety (site II). The intramolecular proton transfer, which leads to the most stable Al(III)-AHTC MHL2+ complex, has not been considered by the experimentalists. The experimental structure was found to be unfavorable in our calculations in both gas phase and water solution. All the semiempirical results are in perfect agreement with the ab initio calculations. So, we suggest that the experimental assignments should be revised, taking into account the results obtained in the present study. PMID- 9724563 TI - Intracellular or intercellular localization of the polar pathway of penetration across stratum corneum. AB - A pharmacokinetic hypothesis of stratum corneum with two parallel pathways, lipophilic and porous hydrophilic, is not well documented yet. Still questionable is the localization of the pores, and the present experiments were designed to elucidate the contribution of extracellular lipids and intracellular keratin to the structure of this pathway. Percutaneous penetration of baclofen, a model zwitterion, was studied in vitro using human cadaver skin. Aqueous or ethanolic saturated solutions of the drug (Cs = 4.6 and 0.4 mg/mL, respectively) were applied on the skin that was pretreated with: methanol/chloroform (Me/Ch) or acetone-chloroform (Ac/Ch) (1:1) mixtures, or with these solvents followed by 0.2% solution of sodium lauryl sulfate (SLS). As controls, baclofen penetration through the intact full-thickness skin was determined, and the fluxes were 0.18 +/- 0.08 and 0.14 +/- 0.07 microg/cm2/h for aqueous and ethanolic solutions, respectively. When Me/Ch was used for 1 h, an expected increase of the penetration was observed, but the lag time, Tlag, was still nearly 20 h. When the less polar mixture, Ac/Ch, was used, no flux enhancement was observed, and with ethanol as the vehicle, decreased penetration was even noted. No effect on baclofen penetration was observed when SLS was used for 1 h after delipidization of the skin was done with either the Me/Ch or Ac/Ch mixture. The results suggest that the polar pathway may be located intercellularly and comprises aqueous regions surrounded by polar lipids, which create the walls of such microchannels. PMID- 9724564 TI - Polymer erosion and drug release characterization of hydroxypropyl methylcellulose matrices. AB - Polymer erosion of matrices of similarly substituted hydroxypropyl methylcellulose (HPMC) polymers was examined, and drug release in terms of diffusion and erosion contributions was characterized, focusing on matrices containing either polymer alone or a drug content of 25% level with no added excipients. A novel approach was utilized to separate diffusional and erosional contributions to drug release. Diffusional drug release was determined by fitting release data versus (time)0.45, and the drug release due to erosion was quantified by subtracting the percent predicted for diffusional drug release from the total drug release at each specific time point. Drug release resulting from polymer erosion was linear versus time and was found to be a function of the number average molecular weight of the polymer. In contrast, diffusional release rates were comparable for all HPMC grades studied and, thus, were independent of number average molecular weight of the polymers studied. Under stirring conditions of 10-100 rpm as well as static condition, the detachment of individual polymer chains at the matrix surface occurred at a faster rate relative to diffusion away from the matrix surface. The erosion study indicated that polymer diffusion of the HPMC polymer chains through the aqueous diffusion layer was the rate-limiting step for polymer erosion. In general, polymer erosion was found to be inversely related to the polymer number average molecular weight. A scaling law was used to relate polymer erosion rate with the respective polymer number average molecular weight. Similar relationships were obtained for matrices with and without drug at a stirring rate of 100 rpm. PMID- 9724565 TI - Use of Fourier transform infrared (FTIR) spectroscopy to follow the adsorption of heptane and 1,4-dioxane vapors on a zinc oxide surface. AB - Vapor adsorption isotherms of two nonpolar model compounds, heptane and 1,4 dioxane, were determined for a very small particle size zinc oxide (ZnO) powder (median particle size approximately 23 nm) in the lower relative vapor pressure (P/Po) region. The ZnO samples for all adsorption measurements were dried at 400 degrees C for 4 h. A new method, which employed an FTIR spectrometer with a long path gas cell (IR path length of 3.0 m), was developed for the organic vapor adsorption measurements. The amount adsorbed was determined by mass balance. This method allows accurate quantification of organic vapors and is sensitive to very low P/Po values. The heptane and 1, 4-dioxane vapor adsorption isotherms appeared to exhibit the expected Type II behavior. The surface areas obtained for ZnO from BET analyses of the heptane and 1,4-dioxane vapor adsorption isotherms (36.9 and 30.3 m2/g) compared reasonably well to the surface area obtained from BET analysis of the nitrogen vapor adsorption isotherm (32.6 m2/g). The amount of vapor adsorbed by ZnO at P/Po equal to 0.1, in terms of number of moles, was observed to decrease in the order: water10 >> 1,4-dioxane > heptane. It was inferred that, while heptane was only adsorbed via a dipole-induced dipole interaction, 1,4-dioxane was physically adsorbed via an interaction dominated by the oxygen lone-pair orbital. Presumably, this interaction was more comparable to a weak dipole-dipole interaction. These results are consistent with the expected strengths of interaction. PMID- 9724566 TI - Short-term chronoamperometric screening of chlorpromazine-package interactions. AB - A new electroanalytical method has been developed to measure and predict solute sorption interactions with solid surfaces. By maximizing surface-to-volume ratios, this method significantly reduces the study time of drug-package interactions and allows prediction of possible long-term effects. Chronoamperometry experiments were run in 40 microL drops of solution containing drug placed on a solid substrate disk of about 7 mm diameter in a sample cell designed to accommodate a miniaturized three-electrode setup. Logarithmic current signatures obtained by computing Delta(ln i)/Delta(ln t) were used to define the experimental conditions necessary to avoid the kinetic complications of chlorpromazine oxidation in the interpretation of the results of the chronoamperometric analysis. Results of sorption studies of chlorpromazine to glass, polypropylene, high density polyethylene, poly(ethylene terephthalate), ethylene vinyl acetate, and poly(vinyl chloride) are presented. The small volume sorption experiments demonstrated that chlorpromazine interacts most quickly with PVC and HDPE and least with glass and polypropylene. Long term stability tests confirmed these predictions, thereby indicating that the small volume method makes drug-package interaction studies feasible in early development. The generation and analysis of Delta(ln i)/Delta(ln t) signature curves extends the usefulness of the electroanalytical method to other systems by accurately identifying the appropriate time domains for steady state or Cottrell behavior. PMID- 9724567 TI - Comparison of the effects of potential parenteral vehicles for poorly water soluble anticancer drugs (organic cosolvents and cyclodextrin solutions) on cultured endothelial cells (HUV-EC). AB - The effect of dilution of parenteral vehicles (organic cosolvent and 0.1 M cyclodextrin solutions) on cultured endothelial cells (HUV-EC) were compared in vitro. Cell morphology was observed by phase contrast light microscopy and cell viability by measuring 3-[4, 5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) reduction or intracellular lactate dehydrogenase (LDH) activity and total protein. Disruption of the HUV-EC monolayer was observed at dilutions of 1 in 20 for the melphalan and PEP cosolvents, 1 in 100 for an investigational drug cosolvent, and 1 in 10 for 0.1 M dimethyl-beta-cyclodextrin. In comparison, 0.1 M SBE7M- and HP-beta-cyclodextrin caused only minor disruption at a 1 in 5 dilution. MTT reduction, intracellular LDH, and total protein were decreased following exposure to 1 in 10 dilution of the melphalan cosolvent. For other test solutions, intracellular LDH activity and total protein were measured, and reductions were observed following exposure to 1 in 10, 20, and 50 dilutions of the investigational drug cosolvent and 1 in 5 dilution of DM-beta-cyclodextrin (0.1 M). At a dilution of 1 in 10, no delayed toxicity was observed for cosolvents or cyclodextrin solutions. Hence, 0.1 M SBE7M- or HP-beta-cyclodextrin formulations may be less damaging to the venous endothelium at the site of injection than organic cosolvent formulations. PMID- 9724568 TI - Liposomes, a potential immunoadjuvant and carrier for a cryptococcal vaccine. AB - Mice immunized with a cryptococcal culture filtrate antigen (CneF) emulsified in complete Freund's adjuvant (CFA) develop an anticryptococcal cell-mediated immune response (CMI). CMI is detected by delayed-type hypersensitivity (DTH) reactions and by enhanced clearance of Cryptococcus neoformans from infected tissues. The objective of this research was to evaluate anticryptococcal DTH reactivity and clearance of cryptococci from groups of mice immunized with CneF encapsulated into liposomes (CneF-liposome) and compare the results to results from mice immunized with CneF-CFA. CBA/J mice were injected subcutaneously with vaccines or control formulations (saline-liposome or saline-CFA). Six days later the mice were footpad tested to assess their DTH response to CneF or the animals were challenged intravenously with 10(5) viable C. neoformans to determine clearance of infection. Clearance was evaluated 7 days later by enumeration of cryptococcal colony forming units (CFU) in lungs, spleens, livers, and brains of the infected mice. The CneF-liposome formulation induced a positive anticryptococcal DTH response and elicited increased clearance of C. neoformans from tissues as compared to mice treated with saline-liposome. Even though the CneF-liposome preparation did not induce as strong of a DTH response or as much protection as did CneF-CFA, our results indicate that liposomes are promising carriers for immunization with cryptococcal antigen and that such immunization can provide some protection to subsequent infection with C. neoformans. PMID- 9724569 TI - Preparation and in vitro characterization of gentamycin-impregnated biodegradable beads suitable for treatment of osteomyelitis. AB - A new method for preparing poly(L-lactide) (PLA) biodegradable beads impregnated with an ionic aminoglycoside, gentamycin, is described. The process employs hydrophobic ion pairing to solubilize gentamycin in a solvent compatible with PLA, followed by precipitation with a compressed antisolvent (supercritical carbon dioxide). The resulting precipitate is a homogeneous dispersion of the ion paired drug in PLA microspheres. The microspheres are approximately 1 microm in diameter and can be compressed into beads (3-6 mm in diameter) strung on surgical sutures for implantation. The bead strings exhibit no significant change in release kinetics upon sterilization with a hydrogen peroxide plasma (Ster-Rad). The kinetics of gentamycin release from the PLA beads are consistent with a matrix-controlled diffusion mechanism. While nonbiodegradable poly(methyl methacrylate) (PMMA) beads initially release gentamycin in a similar manner, the drug release from PMMA ceases after 8 or 9 weeks, while the PLA beads continue to release drug for over 4 months. Moreover, only 10% of the gentamycin is released from the PMMA beads, while PLA beads release more than 60% of their load, if serum is present in the release medium. The PLA system displays improved release kinetics relative to PMMA, is biodegradable, is unaltered by gas sterilization, can be used for a range of antibiotics, and can be manipulated without disintegration. These are all desirable properties for an implantable drug delivery system for the prevention or treatment of osteomyelitis. PMID- 9724570 TI - Effects of freeze-dry processing conditions on the crystallization of pentamidine isethionate. AB - The results of this study show that pentamidine isethionate (PI) can exist in at least four crystalline forms-three anhydrates designated as forms A, B, and C, and a trihydrate. Form C is the high-temperature modification, produced by heating forms A, B, and the trihydrate above 130 degrees C and cannot be produced under actual lyophilization conditions. The crystal forms of PI present after freeze-drying depend on the initial solution concentration and the thermal history of freezing. At low concentrations of PI (4% and less), form A is observed regardless of freezing method. At a higher concentration (10%), the crystal forms observed are a function of the freezing method. Three freezing methods were used to effect different cooling rates: (1) cooling on the shelf to 2 degrees C and holding for 3 h prior to decreasing the temperature to -45 degrees C, (2) directly cooling on the shelf from room temperature to -45 degrees C, and (3) dipping the vials in liquid nitrogen. The results show that form A, form B, or a mixture of both forms are present in the freeze-dried solid depending upon whether the trihydrate crystallizes during freezing or not. Since form B can only be produced by dehydration of the trihydrate at low temperature, the presence of this form in the freeze-dried powders depends on the nucleation and growth of the trihydrate during freezing. Photostability studies have demonstrated marked differences between freeze-dried solids frozen under different conditions. The results underscore the importance of recognizing that seemingly subtle differences in processing conditions can have a significant impact on critical quality attributes of freeze-dried products. PMID- 9724571 TI - Flux enhancement effects of ionic surfactants upon passive and electroosmotic transdermal transport. AB - This study focused upon the enhancement effects of ionic surfactants upon passive and electroosmotic transdermal flux. The first phase of the study involved validating theories relating surface properties of a membrane to electroosmotic solvent flow under appropriate experimental conditions using a synthetic model membrane (stack of 50 Nuclepore membranes). Numerical solutions to the Poisson Boltzmann equation and the equations of fluid motion served as the theoretical basis for the experimental studies. Important outcomes of the model membrane studies were that electroosmotic solvent flow velocity was enhanced by the addition of an anionic surfactant, sodium dodecyl sulfate, and reversed by the addition of a cationic surfactant, dodecyltrimethylammonium bromide. The effective membrane pore wall surface charge densities were determined under a variety of experimental conditions. Adsorption of dodecyl sulfate to the pore wall increased the net negative charge on the pore wall. A reversal of the net pore wall surface charge density resulted from the adsorption of dodecyltrimethylammonium. The interrelationship between electroosmosis, surfactant adsorption, and ionic strength was also evaluated. The second phase of the study was an investigation of the effects of sodium dodecyl sulfate upon the transport of neutral polar permeants through human epidermal membrane (HEM). Fluxes of [14C]urea and [3H]sucrose were simultaneously measured across HEM samples under passive and 250 mV conditions; flux measurements were made before, during, and after HEM exposure to sodium dodecyl sulfate. A systematic analysis of the experimental data made it possible to elucidate the specific contributions of sodium dodecyl sulfate and the applied electric potential to the overall flux enhancement. Sodium dodecyl sulfate enhanced the intrinsic passive permeability of the HEM, and it also enhanced the contribution of electroosmosis to the flux during iontophoresis. PMID- 9724572 TI - Absolute bioavailability and dose proportionality of BMS-181885, an antimigraine agent, following the administration of single intranasal doses to cynomolgus monkeys. PMID- 9724573 TI - Effect of cilastatin on renal handling of vancomycin in rats. AB - To provide insights into the possibility of reducing the nephrotoxicity of vancomycin (VCM) by cilastatin, the effect of cilastatin on the renal handling of VCM, as well as on glomerular filtration rate (GFR) and plasma protein binding of VCM, were studied using rats. After a bolus intravenous (iv) dose of VCM (100 mg/kg), concomitant cilastatin administration (100 mg/kg, iv) resulted in a significant increase in the total VCM clearance and significant decrease in the kidney uptake clearance of VCM, defined as kidney VCM concentration vs AUC ratio. Moreover, after a 3-h continuous iv infusion of VCM (18 or 90 mg/h/kg), significant decrease in the kidney uptake clearance of VCM was observed with concomitant cilastatin iv infusion (300 mg/h/kg). On the other hand, GFR and VCM plasma protein binding did not show any significant change with cilastatin. From the observation that cilastatin decreased the kidney uptake clearance of VCM and enhanced its urinary excretion, it was suggested that cilastatin inhibited the reabsorption of VCM in the renal proximal tubular cells. Thus, it may be possible that cilastatin alleviates the nephrotoxicity of VCM due to reduced accumulation and accelerated renal excretion of VCM. PMID- 9724574 TI - The Microwave Spectrum of n-Butyraldehyde Oxime. AB - The microwave spectrum of n-butyraldehyde oxime was observed in the frequency region 26.5-40 GHz. Four rotational conformers were found to exist in the gas phase; among these, two conformers belonged to the (E)-geometrical isomer and the other two to the (Z)-geometrical isomer. The microwave spectrum attributed to one of these two rotational conformers of (E)-butyraldehyde oxime was analyzed, and its rotational constants for the ground vibrational state were determined: A = 15883(379), B = 1269.97(1), C = 1251.60(1) MHz. The conformational structure of the molecule is discussed, referring to the rotational constants obtained and the ab initio molecular orbital calculation. Copyright 1998 Academic Press. PMID- 9724575 TI - First High-Resolution Study of the 13CHD3 Infrared Spectrum: Rotational Analysis of the Ground State and the Fundamentals nu5 and nu3/nu6. AB - The rotational structure in three fundamental bands nu3, nu5, and nu6 of 13CHD3 has been analyzed. As this study evidently is the first high-resolution investigation on the spectrum of this molecule, the determination of the ground state rotational constants was the necessary first step. The nu5 band (1290.536 cm-1) could be treated as an isolated band whereas it was necessary to analyze the bands nu3 (994.713 cm-1) and nu6 (1030.243 cm-1) simultaneously because of very strong interactions. The total number of assigned lines was 1116. The spectral region 850-1530 cm-1 was measured on a Fourier transform spectrometer and the 13C species was in natural abundance in CHD3. Copyright 1998 Academic Press. PMID- 9724576 TI - High Frequency Transitions in the Rotational Spectrum of SO2. AB - A large number of rotational transitions of 32S16O2, 34S16O2, and 32S18O16O have been measured in the mm-, submm-, and terahertz ( approximately 1 THz) spectral regions. These data sets have been combined with all previously measured SO2 microwave and selected far infrared data to obtain a highly precise set of ground state rotational constants for these isotopomers. The rotational constants for the three isotopomers are in MHz as follows: Parameter32S16O234S16O232S18O16O A60778.54977 (44)58991.18295 (51)59101.1690 (27) B10318.07348 (7)10318.50993 (9)9724.64284 (56) C8799.703399 (70)8761.302481 (97)8331.56018 (51) Centrifugal distortion constants up to P10 are included in the fit. A frequency listing of all the data used in the frequency range between about 7 GHz and 1 THz is included. Copyright 1998 Academic Press. PMID- 9724577 TI - On the 5d 1Pig --> 2 (1)Sigma+u and 5d 1Pig --> C 1Piu Fluorescence in 7Li2. AB - Following excitation of the 5d 1Pig Rydberg state of 7Li2 by optical-optical double resonance, fluorescence has been observed in the infrared region to the 2 (1)Sigma+u and C 1Piu states. Analysis of high-resolution Fourier transform spectra yields term energies and rotational constants of the lowest seven vibrational levels of the inner well of the 2 (1)Sigma+u "double minimum" state. The equilibrium term value and dissociation energy have been determined to be Te = 30101.45 +/- 0.12 cm-1 and De = 5621.3 +/- 0.2 cm-1. The v = 0 and 1 levels of the C 1Piu state have been analyzed, resulting in new values of Te = 30551.0 +/- 0.1 cm-1 and De = 7773.3 +/- 0.2 cm 1. Copyright 1998 Academic Press. PMID- 9724578 TI - Fourier Transform Spectroscopy of Carbonyl Sulfide from 4800 to 8000 cm-1 and New Global Analysis of 16O12C32S. AB - We have measured the FT spectrum of natural OCS from 4800 to 8000 cm-1 with a near Doppler resolution and a line-position accuracy between 2 and 8 x 10(-4) cm 1. For the normal isotopic species 16O12C32S, 37 vibrational transitions have been analyzed for both frequencies and intensities. We also report six bands of 16O12C34S, five bands of 16O13C32S, two bands of 16O12C33S, and two bands of 18O12C32S. Important effective Herman-Wallis terms are explained by the anharmonic resonances between closely spaced states. As those results complete the study of the Fourier transform spectra of natural carbonyl sulfide from 1800 to 8000 cm-1, a new global rovibrational analysis of 16O12C32S has been performed. We have determined a set of 148 molecular parameters, and a statistical agreement is obtained with all the available experimental data. Copyright 1998 Academic Press. PMID- 9724579 TI - Free Jet Absorption Millimeter Wave Spectrum of Pyrrolidine: Assignment of a Second, Equatorial, the Most Stable Conformer. AB - The equatorial conformer of pyrrolidine has been discovered while investigating the rotational free jet spectrum of the pyrrolidine-water adduct. It is more stable than the axial species, previously assigned with conventional microwave spectroscopy (W. Caminati, H. Oberhammer, G. Pfafferott, R. R. Filgueira, and C. H. Gomez, 1984. J. Mol. Spectrosc., 106, 217-266). The assignment of the equatorial conformer was missed in that microwave investigation because of the accidentally zero or almost zero value of its ua dipole moment component; its low J uc-type transitions, very weak in the room temperature spectrum, are the strongest lines in the jet. Copyright 1998 Academic Press. PMID- 9724580 TI - The S1(1A1)-S0(1A1) Electronic Transition of Jet-Cooled o-Difluorobenzene. AB - A detailed study of the S1(1A1)-S0(1A1) transition of jet-cooled o difluorobenzene has been completed using the two techniques of laser-induced fluorescence excitation and dispersed, single vibronic level fluorescence spectroscopy. Analysis of over 60 dispersed fluorescence spectra resulted in both the assignment of 22 excited state vibrational frequencies and the confirmation of 23 ground state frequencies. The spectrum is dominated by Franck-Condon activity in totally symmetric vibrations with long progressions in the ring breathing mode, nu9. By analogy with benzene and the para- and meta-substituted isomers, two vibronic coupling mechanisms are postulated to be responsible for the wealth of weaker symmetry-forbidden structure that has been observed. Single quantum changes in b2 vibrations are postulated to appear due to first order vibronic coupling to a higher lying B2 electronic state. Combinations of b1 and a2 modes are postulated to appear from second order vibronic coupling to an A1 electronic state. This second order coupling causes a pronounced Duschinsky mixing among excited state b1 and a2 modes with respect to their ground state counterparts. Franck-Condon factors are calculated for the a1 progression-forming modes, anharmonic contributions are evaluated, one strong Fermi resonance is identified and analyzed, and the Duschinsky rotation matrix elements are evaluated for the most strongly affected modes, nu17 and nu18. Several transitions in the oDFB-oDFB van der Waals dimer and oDFB-Ar complex are also assigned in the spectrum. Copyright 1998 Academic Press. PMID- 9724581 TI - Rotational Energy Levels and Line Intensities for 2S+1Sigma-2S+1Sigma Transitions in an Open-Shell Diatomic Molecule Weakly Bonded to a Closed-Shell Partner. AB - This paper concerns rotational energy levels and line intensities for electronic, vibrational, and microwave transitions in an open-shell complex consisting of an open-shell diatomic molecule and a closed-shell partner. The electronic state of the open-shell diatomic fragment is a 2S+1Sigma state, where S >/= 12, the close shell partner could be a rare gas atom or a diatomic molecule or a planar polyatomic molecule. We are considering a near-rigid rotor model for a nonlinear complex, taking into account thoroughly all effects of the electron spin and the quartic centrifugal distortion correction terms. The total Hamiltonian is expressed as H=Hrot+Hsr+Hss+Hcd+Hsrcd+Hsscd. We have derived all the nonvanishing matrix elements of the Hamiltonian operators in the molecular basis set. The rotational energy levels are calculated by numerical diagonalization of the total Hamiltonian matrix for each J value. The nonvanishing matrix elements of the electric dipole moment operator are derived in the molecular basis set for electronic, vibrational, and microwave transitions within the complex. Expectation values of the quantum numbers and of the parities of the rotational states are derived in the molecular basis set. Relative intensities of the allowed rotational transitions, expectation values of the quantum numbers and the parities are calculated numerically in the space of the eigenvectors obtained from diagonalization of the Hamiltonian matrix. The formalism and the computer program of this paper are considered as extensions to our previous work [W. M. Fawzy and J. T. Hougen, J. Mol. Spectrosc. 137, 154-165 (1989); W. M. Fawzy, J. Mol. Spectrosc. 160, 84-96 (1993)] and are expected to be particularly useful for analyzing and fitting high-resolution spectra of weakly bonded oxygen complexes. A brief discussion of the Hamiltonian operators, the matrix elements, and the computer program is given. Copyright 1998 Academic Press. PMID- 9724582 TI - Spectroscopy and Lifetimes of the 1(2)Piu State of Na+2: A New Comparison between Theory and Experiment. AB - A detailed theoretical study of the spectroscopy of the 1(2)Piu state of Na+2 is presented, including the determination of Dunham coefficients from recent effective potential calculations of the Na+2 molecular potential curves. We emphasize the discrepancies existing among several experiments and among several theoretical models available in the literature. The radiative lifetime of the rovibrational levels of the 1(2)Piu state are also computed and are found in agreement within 20% with experiment. Copyright 1998 Academic Press. PMID- 9724583 TI - High-Resolution Infrared Fourier-Transform Spectroscopy and Rotational Constants of the nu4 Band of BrNO2. AB - For the first time, high-resolution infrared gas-phase absorption spectra of the BrNO2 molecule were recorded using a Fourier-transform spectrometer. In this paper, the nu4 bands of the 79BrNO2 and 81BrNO2 isotopomers around 1670 cm-1 are investigated. Although the spectra are highly congested, rotational and centrifugal distortion constants for the ground and v4 = 1 states of 79BrNO2 and 81BrNO2 were determined. The results show that BrNO2 is a planar molecule of C2nu symmetry and confirm predictions from a recent ab initio study. Copyright 1998 Academic Press. PMID- 9724584 TI - Lifetime Measurements of the C3Delta State of TiS and the E3Pi State of TiO. AB - Lifetime measurements of the C3Delta electronic state of the titanium sulfide molecule and of the E3Pi electronic state of the titanium oxide molecule have been performed using population probing with resonant two-photon ionization in a molecular beam. The lifetimes for the TiS C3Delta1 state were found to be 243 +/- 7 ns, 278 +/- 9 ns, and 286 +/- 15 ns for the v = 0, v = 1, and v = 2 levels, respectively. The lifetime for the TiO E3Pi0 state was found to be 4.9 +/- 0.2 us for the v = 0 level. Copyright 1998 Academic Press. PMID- 9724585 TI - Infrared Laser Velocity Modulation Spectroscopy of SiCl+(X1Sigma+) up to v = 6. AB - Many fundamental and hot band absorption lines of 28Si35Cl+, 28Si37Cl+, 29Si35Cl+, and 30Si35Cl+ have been detectedbetween 630 and 700 cm-1 in a SiCl4/He discharge using diode laser velocity modulation spectroscopy. The datahave been fitted to give seven mass independent coefficients Ukl. The derived spectroscopic constants for 28Si35Cl+ include omegae = 678.24316(18) cm-1 and Be = 0.2870288(14) cm-1. The equilibrium internuclear distance is re =1.9439105(46) A. The equilibrium dissociation energy calculated from Dunham's expanded Morse potential is De =49 431(57) cm-1. Copyright 1998 Academic Press. PMID- 9724586 TI - High-Resolution Study of the X22Pi3/2 --> X12Pi1/2 Fine Structure Transitions of PbF and PbCl. AB - The near-infrared emission spectra of the X22Pi3/2 --> X12Pi1/2 fine structure transitions of PbF and PbCl have been investigated by high-resolution Fourier transform spectrometry. Despite the large fine structure splitting, the bands can be equally well analyzed with Hund's case (a) or case (c) formalisms. Accurate rotational constants for the v = 0 vibrational levels of 208PbF and 208Pb35Cl and hyperfine structure constants for 207PbF have been derived. Copyright 1998 Academic Press. PMID- 9724587 TI - High-Resolution Infrared Spectrum of Monoiodoacetylene Between 2000 and 3000 cm 1. AB - The high-resolution infrared spectrum of monoiodoacetylene measured with a Bruker IFS 120 HR Fourier transform spectrometer in the spectral range of 2000-3000 cm-1 has been studied in detail. The strongest bands observed in the wavenumber region investigated are the C-C stretching fundamental nu2 (2034-2082 cm-1) and the accompanying hot bands of the types nu2 + nun - nun, where n = 3, 4, or 5, associated with the lowest stretching state v3 = 1 and with the low-lying bending states v4 = 1 and v5 = 1, 2, respectively. The combination bands of the types nu2 + nun, where n is as above, starting direct from the ground state have also been observed. In addition, the weak overtone band 2nu2 (4081-4121 cm-1) has been measured to study the anharmonicity of the C-C stretching mode. The new, more accurate values for the ground state rotational constants have been derived by combining the ground state combination differences calculated from our IR data available in the present work as well as in our previous investigations concerning the HCCI molecule to the accurate MW transitions from the literature. The rotational structures of the overtone level v2 = 2 and the combination levels v2 = vn = 1, where n is as above, have been investigated by analyzing the observed spectra with a model including various Fermi- and l-type resonances. As a result, the values for the harmonic frequency omega02 and the anharmonicity constants x22, x23, x24, and x25 are determined. Copyright 1998 Academic Press. PMID- 9724588 TI - Fourier Transform Emission Spectroscopy of TaO. AB - The emission spectrum of TaO, excited in a tantalum hollow cathode lamp, has been observed at high resolution using a Fourier transform spectrometer. In addition to previously known transitions, a number of new bands have been identified and assigned as belonging to two new electronic transitions: H2Pi1/2-X2Delta3/2 and K2Phi5/2-X2Delta3/2. A rotational analysis ofthe 0-0 and 0-1 bands of the H2Pi1/2 X2Delta3/2 transition and of the 0-1, 1-2, 0-0, 1-0, and 2-1 bands of theK2Phi5/2 X2Delta3/2 transition has been carried out, providing the following equilibrium constants for the ground X2Delta3/2 state:omegae = 1028.9060(15) cm-1, omegaexe = 3.58928(66) cm-1, Be = 0.40289737(139) cm-1, alphae = 0.00185445(83) cm-1, andre = 1.6873430(29) A. The principal molecular constants for the H2Pi1/2 state are T00 = 20 634.32758 (40) cm-1,B0 = 0.3766867(31) cm-1, and r0 = 1.7450604(72) A, while the equilibrium constants for the K2Phi5/2 state areomegae = 905.4549(15) cm-1, omegaexe = 3.67601(64) cm-1, Be = 0.37965102(36) cm-1, alphae = 0.00189370(21) cm-1, andre = 1.7382343(8) A. Although the H2Pi1/2 and K2Phi5/2 states have been observed previously in matrix isolation experiments, our work on these states is the first in the gas phase. Copyright 1998 Academic Press. PMID- 9724589 TI - Modified Morse Potential for Diatomic Molecules. AB - We present a diatomic potential which closely resembles the standard Morse function but incorporates additional flexibility for fitting experimental vibrational energy-gap data. This flexibility is accommodated by introducing a continuously variable radially dependent change in the exponent of the Morse function, which in practice is adequately realized via a relatively small number of constant parameters. As an illustration, the method is applied to calculate the quantum vibrational levels of the X1Sigma+g ground electronic state of the N2 molecule. Copyright 1998 Academic Press. PMID- 9724590 TI - The H1Sigma+g (v = 0 and 1) and EF1Sigma+g (v = 28 and 32) States of D2: Term Values and Fluorescence Lifetimes. AB - The extreme ultraviolet-visible double resonant excitation method was applied to the gerade Rydberg states of D2 molecules. Tunable coherent extreme ultraviolet radiation near 103 nm prepared D2 in the B1Sigma+u (v = 7, J) state. Visible laser light subsequently brought them to the H1Sigma+g (v = 0 and 1), EF1Sigma+g (v = 28 and 32), and GK1Sigma+g (v = 2 and 5) states. Our term values for the GK1Sigma+g state were in complete agreement with previous values, while deviations beyond the experimental error were found for the H1Sigma+g (v = 1) and EF1Sigma+g (v = 28 and 32) states. The fluorescence lifetimes, measured under a collision-free environment for the first time, were in reasonable agreement with ab initio calculations. Copyright 1998 Academic Press. PMID- 9724591 TI - Methyl Chloride nu5 Region Lineshape Parameters and Rotational Constants for the nu2, nu5, and 2nu3 Vibrational Bands. AB - Fourier spectra of methyl chloride have been obtained at ambient laboratory temperature under self-broadening conditions. Five thousand two hundred and sixty rovibrational transitions of the (nu2, nu5, 2nu3)-vibrational-band triad have been assigned, and new ground and excited state rotational constants for CH335Cl and CH327Cl have been determined. The nu5 and nu2 vibrational-band intensities are determined to be, respectively, 42(1.2) and 31.7(0.9) cm-2 atm-1. Self broadening coefficients of individual rovibrational lines are determined for the nu5 band of CH335Cl and CH337Cl at 296 K. The self-broadening coefficients peak broadly close to the maximum in the Boltzmann rotational population; no specific trends of the broadening coefficients are observed with the rotational quantum number K. With the exception of the RQ(J, 0) branch, the 1260-1650 cm-1 spectral region can be well modeled by the superposition of overlapping Voigt line profiles. To adequately model the densely packed RQ(J, 0) branch, however, we included collisional line mixing with A+ to A- in addition to A- to A- and A+ to A+ |DeltaJ| >/= 1, DeltaK = 0 collisional transitions allowed. Copyright 1998 Academic Press. PMID- 9724592 TI - Rotational Spectra of the Isotopic Species of Chloroform: Experimental and Ab Initio Structures. AB - The millimeter-wave spectra of three different samples of chloroform (CHCl3, CDCl3, and 13CHCl3) have been measured between 145 and 470 GHz which corresponds to J values between 22 and 70. We report accurate rotational and centrifugal distortion constants for the ground vibrational states of 11 isotopic species. The experimental ro, rs, rIepsilon, rrhom, and rz structures have been determined using the determined rotational constants. The structure has also been calculated ab initio at the SCF, MP2, RQCISD, and B3LYP levels using triple zeta polarized basis sets. The experimental results are found in excellent agreement with the ab initio predictions. An approximate equilibrium structure has been obtained by combining the experimental results and the ab initio calculations: re(C-H) = 1.080 (2) A, re(C-Cl) = 1.760 (2) A, and anglee(HCCl) = 108.23 (2) degrees. Copyright 1998 Academic Press. PMID- 9724593 TI - The Pure Rotational Spectra of TiO(X3Delta) and TiN(X2Sigma+). AB - The pure rotational transitions in the 220-460 GHz spectral range of TiO (X3Deltar) and TiN(X2Sigma+) have been observed using a source-modulated submillimeter-wave spectrometer. In addition, the two lowest rotational transitions of TiO at 63 and 94 GHz have been observed using molecular beam pump/probe microwave optical double resonance (PPMODR) technique. An improved set of spectroscopic parameters for TiO(X3Delta) was generated by combining the new PPMODR and submillimeter-wave absorption measurements. Similarly, an improved set of spectroscopic parameters for TiN(X2Sigma+) was generated by combining the new submillimeter-wave absorption measurements with the previously recorded PPMODR measurements (D. A. Fletcher, C. T. Scurlock, K. Y. Jung, and T. C. Steimle, 1993, J. Chem. Phys. 99, 4288). The determined Lambda-type doubling parameter for the TiO(X3 Delta) is compared with the theoretical model. Copyright 1998 Academic Press. PMID- 9724594 TI - Analysis of the Rotational Spectra of SiH3CN and Its Isotopomers: Experimental and Ab Initio Determinations of the Dipole Moment and the Structure. AB - The ground state rotational spectra of SiH3CN and its 29Si, 30Si, 13C, 15N, d1, d2, and d3 isotopic species have been measured by Fourier transform microwave spectroscopy and by millimeterwave spectroscopy. Accurate rotational, centrifugal distortion, and 14N and D nuclear quadrupole coupling constants have been derived. The dipole moment of the parent species has also been measured, u = 3.4400(42) D. The structure, force field, dipole moment, and nuclear quadrupole coupling constants have been calculated ab initio at the SCF, MP2, and B3LYP levels using triple zeta polarized basis sets. The experimental ro, rs, and rz structures have been determined. An approximate equilibrium structure has been obtained by combining the experimental results and the ab initio calculations: re(C&tbond;N) = 1.159 A, re(Si&sbond;C) = 1.848 A, re(Si&sbond;H) = 1.470 A, and angle(HSiC) = 107.4 degrees. Copyright 1998 Academic Press. PMID- 9724595 TI - LIF Study of the b1Sigma+(b0(+)) left and right arrow X3Sigma-(X10(+),X21) Transitions of SbH and SbD. AB - The b1Sigma+(b0(+)) left and right arrow X3Sigma-(X10(+),X21) transitions of SbH and SbD have been studied by laser-induced fluorescence measurements. Ground state SbH(SbD) radicals were generated in a fast-flow system by reaction of microwave-discharged hydrogen (deuterium) with antimony vapor. Vibrational levels v = 0 and 1 of the b1Sigma+ state were excited with a pulsed dye laser. In addition to five electric dipole branches SR, QP, QR, QQ, and OP, weak magnetic dipole branches PQ were observed in the 0-0 bands of the b1Sigma+ <-- X3Sigma- transitions. Rotational and vibrational constants of the X3Sigma- and b1Sigma+ states were deduced from high-resolution excitation spectra. Time-resolved measurements of the fluorescence decay and measurements of relative line and band intensities allowed determination of the radiative lifetime of the b1Sigma+ state of SbH (tau = 173 +/- 3 us) and of the electric and magnetic dipole transition moments u0 = +/-0.036 D, u1 = -/+0.058 D, |M| = 1.62 Bohr magnetons. Copyright 1998 Academic Press. PMID- 9724596 TI - Frequencies of Optically Pumped Sub-Doppler Far-Infrared Laser Lines of Methanol. AB - Sub-Doppler signals from the gain curves of four lines of a transversely pumped far-infrared methanol laser were used to stabilize the frequencies of the laser. The frequencies of four of the strongest FIR laser lines (70, 119, 123, and 570 um) were measured. Lasers locked to these lines have a frequency reproducibility of a few parts in 10(8) and an uncertainty of less than 30 kHz. Copyright 1998 Academic Press. PMID- 9724598 TI - The Infrared Spectrum of H+3 Revealed. PMID- 9724597 TI - The Low-Lying States of He2. AB - The near-infrared emission spectrum of He2, excited in a Be hollow cathode discharge, has been recorded at high resolution using a Fourier transform spectrometer. The c3Sigma+g-a3Sigma+u (0-0, 1 1, 2-2, 1-0, and 2-1) and C1Sigma+g-A1Sigma+u (0-0 and 1-1) transitions have been observed in the 9000-15 000 cm-1 spectral region. A global analysis of the six lowest excited states of He2 (c3Sigma+g, b3Pig, a3Sigma+u, C1Sigma+g, B1Pig, and A1Sigma+u) was carried out by combining our measurements with previously reported infrared data for the b3Pig-a3Sigma+u system [S. A. Rogers et al., Mol. Phys. 63, 901 (1988)] and with laser measurements for the B1Pig-A1Sigma+u transition [H. Solka et al., Mol. Phys. 60, 1179 (1987)]. To account for the fine structure in the a3Sigma+u state, high precision r.f. measurements were included in the global fit. A consistent set of improved molecular constants was derived for the c3Sigma+g (v = 0, 1, and 2), b3Pig (v = 0 and 1), a3Sigma+u (v = 0, 1, and 2), C1Sigma+g (v = 0 and 1), B1Pig (v = 0 and 1), and A1Sigma+u (v = 0 and 1) levels. A study of the vibrational dependence of these constants was also performed, leading to the equilibrium parameters for the six electronic states. Copyright 1998 Academic Press. PMID- 9724599 TI - The A2Sigma+-X2Pii (0, 1) Band and Reanalyses of the Blue and Ultraviolet Transitions of AgO. PMID- 9724601 TI - Fibrous proteins. PMID- 9724602 TI - Why fibrous proteins are romantic. AB - Here I give a personal account of the great history of fibrous protein structure. I describe how Astbury first recognized the essential simplicity of fibrous proteins and their paradigmatic role in protein structure. The poor diffraction patterns yielded by these proteins were then deciphered by Pauling, Crick, Ramachandran and others (in part by model building) to reveal alpha-helical coiled coils, beta-sheets, and the collagen triple helical coiled coil-all characterized by different local sequence periodicities. Longer-range sequence periodicities (or "magic numbers") present in diverse fibrous proteins, such as collagen, tropomyosin, paramyosin, myosin, and were then shown to account for the characteristic axial repeats observed in filaments of these proteins. More recently, analysis of fibrous protein structure has been extended in many cases to atomic resolution, and some systems, such as "leucine zippers," are providing a deeper understanding of protein design than similar studies of globular proteins. In the last sections, I provide some dramatic examples of fibrous protein dynamics. One example is the so-called "spring-loaded" mechanism for viral fusion by the hemagglutinin protein of influenza. Another is the possible conformational changes in prion proteins, implicated in "mad cow disease," which may be related to similar transitions in a variety of globular and fibrous proteins. PMID- 9724603 TI - Supercoiled protein motifs: the collagen triple-helix and the alpha-helical coiled coil. AB - The collagen triple-helix and the alpha-helical coiled coil represent the two basic supercoiled multistranded protein motifs. Originally they were characterized in fibrous proteins, but have been found more recently in a number of other proteins containing rod-shaped domains. Coiled-coil domains are responsible for the oligomerization of proteins, as well as other specific functions, while the triple-helix domains associate to form supramolecular structures and bind a variety of ligands. Both structures were originally solved by fiber diffraction, and recent crystallographic studies on small proteins and peptide models have confirmed the structure and provided molecular details. The differences in the molecular conformations of these two motifs and the interactions stabilizing these conformations are discussed. The molecular structures of both motifs constrain the amino acid sequence to recognizable patterns, requiring the (Gly-X-Y)n repeating sequence for the collagen triple helix and a less stringent heptad repeat requirement (h-x-x-h-x-x-x)n for the coiled-coil domains, where h represents hydrophobic residues. The features and roles of these supercoiled domains in proteins are considered when they are found adjacent in the same protein. PMID- 9724604 TI - The coiled-coil helix in the neck of kinesin. AB - Kinesin is a microtubule-dependent motor protein. We have recently determined the X-ray structure of monomeric and dimeric kinesin from rat brain. The dimer consists of two motor domains, held together by their alpha-helical neck domains forming a coiled coil. Here we analyze the nature of the interactions in the neck domain (residues 339-370). Overall, the neck helix shows a heptad repeat (abcdefg)n typical of coiled coils, with mostly nonpolar residues in positions a and d. However, the first segment (339-355) contains several nonclassical residues in the a and d positions which tend to weaken the hydrophobic interaction along the common interface. Instead, stabilization is achieved by a hydrophobic "coat" formed by the a and d residues and the long aliphatic moieties of lysines and glutamates, extending away from the coiled-coil core. By contrast, the second segment of the kinesin neck (356-370) shows a classical leucine zipper pattern in which most of the hydrophobic residues are buried at the highly symmetrical dimer interface. The end of the neck reveals the structure of a potential coiled-coil "trigger" sequence. PMID- 9724605 TI - Nuclear lamins: their structure, assembly, and interactions. AB - Nuclear lamins are intermediate filament-type proteins that are the major building blocks of the nuclear lamina, a fibrous proteinaceous meshwork underlying the inner nuclear membrane. Lamins can also be localized in the nuclear interior, in a diffuse or spotted pattern. Nuclei assembled in vitro in the absence of lamins are fragile, indicating that lamins mechanically stabilize the cell nucleus. Available evidence also indicates a role for lamins in DNA replication, chromatin organization, spatial arrangement of nuclear pore complexes, nuclear growth, and anchorage of nuclear envelope proteins. In this review we summarize the current state of knowledge on the structure, assembly, and possible functional roles of nuclear lamins, emphasizing the information concerning the ability of nuclear lamins to self-assemble into distinct oligomers and polymers. PMID- 9724606 TI - Hard alpha-keratin intermediate filament chains: substructure of the N- and C terminal domains and the predicted structure and function of the C-terminal domains of type I and type II chains. AB - The quantity of sequence data now available for both Type I and Type II hard alpha-keratin IF proteins makes it possible to analyze their N- and C-terminal domains and ascertain features of likely structural and/or functional importance. The N-terminal domains of both chain types can be divided into acidic (NA) and basic (NB) subdomains, where NA is 29 and 34 residues long, respectively, for Type I and II chains and is located immediately adjacent to the end of the rod domain. NB constitutes the remainder of the N-terminal domain and is about 27 and 70 residues long for the two chain types, respectively. The glycine residue contents, however, are high in NA(I) and NB(II), but low in NA(II) and NB(I). Subdomain NB(II) contains four consecutive nonapeptide quasirepeats of the form GGGFGYRSX. The C-terminal domain of Type I chains, termed C(I), is characterized by a PCX motif repeated 10 times, 7 of them contiguously. From an analysis of the conformation of like peptides from crystal structures it has been shown that this region will probably adopt a polyproline II left-handed helical structure with three residues per turn. In contrast, the C-terminal domain of Type II hard alpha keratin chains (known as C(II)) contains a periodic distribution of hydrophobicities that, together with other predictive techniques, allow its conformation (a twisted four-stranded antiparallel beta-sheet) to be predicted with some degree of confidence. In addition, it is possible to suggest two partners with which this domain will interact. The first is with segment L12 in the rod domain and the second is with another C(II) domain in an antiparallel neighboring molecule. The latter possibility appears most likely. In either case the aggregation would likely serve to stabilize the molecular assembly through the interaction of two beta-sheets via their apolar faces and, in so doing, would position a number of cysteine residues in external positions that would allow them to form a number of covalent disulfide bonds with other molecules. PMID- 9724607 TI - Small proline-rich proteins are cross-bridging proteins in the cornified cell envelopes of stratified squamous epithelia. AB - The cornified cell envelope (CE) is a specialized structure which contributes barrier function to stratified squamous epithelial cells. It is composed of an amalgam of several structural proteins that are rendered insoluble by isopeptide bond crosslinking by transglutaminases. One set of the structural proteins present in CEs of most such epithelia are the small proline rich (SPR) proteins, which are a family of about 12 related structural proteins. We have recovered a large number of peptides containing isopeptide crosslinks, including 236 involving SPR proteins, following proteolysis of CEs isolated from foreskin epidermal tissue and cultured epidermal keratinocytes. Analysis of this database has provided novel information on their function. First, we found that SPRs became crosslinked to many other structural proteins within the CE. Second, multiple glutamine and lysine residues located only on the amino- and carboxy termini of the SPR proteins were involved in crosslinking, so that the two ends are functionally equivalent. Third, the SPRs functioned as cross-bridging proteins, by directly adjoining other CE structural proteins. In the specialized case of the epidermal CE, the SPRs cross-bridged between loricrin. In cultured keratinocytes which make little loricrin and serve as a model for internal stratified squamous epithelia, the SPRs formed extensive cross-bridges among themselves. Thus SPRs are ubiquitous cross-bridging proteins whose differential expression patterns apparently reflect specific barrier requirements of different epithelia. PMID- 9724608 TI - Gly-X-Y tripeptide frequencies in collagen: a context for host-guest triple helical peptides. AB - The collagen triple-helix consists of a repeating (Gly-X-Y)n sequence. In theory, there are more than 400 possible Gly-X-Y triplets, but analysis of sequences from fibrillar and nonfibrillar collagens shows that only a limited set of triplets are found in significant numbers, and many are never observed. The nonrandom frequency of Gly-X-Y triplets makes it practical to experimentally approach the stability of much of the collagen sequence through the study of a limited set of host-guest peptides. In these peptides, individual Gly-X-Y triplets constitute the guest, while the host consists of Gly-Pro-Hyp tripeptides. A set of host guest peptides was designed to contain the most common nonpolar and charged triplets found in collagen. All formed stable triple-helices, with their melting temperature depending on the identity of the guest triplet. While including less than 10% of all possible triplets, the data set covers 50-60% of collagen sequences and provides a starting point for establishing a stability scale to predict the relative stability of important collagen regions, such as the matrix metalloproteinase cleavage site or binding sites. PMID- 9724609 TI - A consensus model for molecular packing of type I collagen. AB - In this review, recent results from X-ray diffraction studies of tendon are used to develop an understanding of the molecular packing of type I collagen in tendon fibrils. These cover the definition of the unit cell as triclinic, the lateral architecture of molecular packing in a fibril and the molecular packing topology of a structure that gives good agreement with X-ray diffraction data. The proposed model is a 1D staggered left handed microfibril; the molecular orientation of the telopeptides indicates that there are interconnections between microfibrils that may explain the difficulty in isolating individual microfibrillar structures. This is the first structure that defines the absolute molecular packing of molecular segments based on X-ray diffraction data. These results are discussed in the light of direct and indirect evidence relating to molecular packing such as mineralization, natural crosslink position, and biomechanical evidence. The ability of the proposed structure to fulfill many of the structural and biochemical criteria point towards the structure providing a basis for a consensus model of collagen packing. PMID- 9724610 TI - Collagen packing in the dogfish egg case wall. AB - The collagen which forms the egg case of the dogfish, Scyliorhinus canicula, is assembled in a three-dimensional network that results in a very resistant capsule. The capsule presumably accomplishes both a protective and a filtering role for the embryo contained within it. In the present work we have obtained electron micrographs of metal-shadowed, deep-etched replicas of the egg case and we have analyzed the micrographs using computer-based Fourier methods. The replicas provide details of the three-dimensional structure that have not been recorded before, allowing us to add particulars to the [1,0,0], [1,1,0], and [0,0,1] views previously obtained from plastic sections (C. Knupp et al., 1996, J. Struct. Biol. 117, 209-221). A molecular packing arrangement consistent with all of the present data is proposed. Although this collagen type is unique and is substantially shorter (45 nm) than other collagens, investigations into its structure may give an insight into related collagen types. The role of this collagen as both a protective structure and one with filtering properties controlling permeability is discussed in terms of other collagens with similar functions. PMID- 9724611 TI - The collagen fibril: the almost crystalline structure. AB - The structure of collagen fibrils has intrigued many investigators over the years. A crystal structure has been available for some time, but the crystal structure has been difficult to reconcile with other observations about collagen fibrils such as their roundness and their growth from paraboloidal tips. Several alternative models recently have been suggested, but none of them fully account for all the data. One recent approach to solving the fibrillar structure is to define specific binding sites on the collagen monomer that direct self-assembly of monomers into fibrils. PMID- 9724612 TI - Fibrillar structure and mechanical properties of collagen. AB - Collagen type I is among the most important stress-carrying protein structures in mammals. Despite their importance for the outstanding mechanical properties of this tissue, there is still a lack of understanding of the processes that lead to the specific shape of the stress-strain curve of collagen. Recent in situ synchrotron X-ray scattering experiments suggest that several different processes could dominate depending on the amount of strain. While at small strains there is a straightening of kinks in the collagen structure, first at the fibrillar then at the molecular level, higher strains lead to molecular gliding within the fibrils and ultimately to a disruption of the fibril structure. Moreover, it was observed that the strain within collagen fibrils is always considerably smaller than in the whole tendon. This phenomenon is still very poorly understood but points toward the existence of additional gliding processes occurring at the interfibrillar level. PMID- 9724613 TI - X-Ray diffraction studies of fibrillin-rich microfibrils: effects of tissue extension on axial and lateral packing. AB - X-ray diffraction of hydrated fibrillin rich microfibrils, in the form of zonular filaments from bovine eyes, demonstrated meridional diffraction peaks indexing on a fundamental periodicity of approximately 56 nm in the relaxed state. The effect of sample extensions of up to 50% in length produced an increase in the axial periodicity of only 4% as judged by alteration of the diffraction peak position of the third meridional order. This effect was shown to be reversible. Further extension to 100% of the tissue rest length caused extensive deterioration in the quality of the diffraction and resulted in a more complex meridional diffraction series, where the fundamental axial periodicity also changed to a length of approximately 80 nm. The fibrillin diffraction image also contains an equatorial diffraction peak that is enhanced upon tissue extension. The measurement of the molecular spacing from the equatorial diffraction profile indicated that the closest approach of molecules gave a broad interference peak of spacing 28 nm, this is nearly twice the molecular diameter as estimated from electron microscopy of dehydrated samples. PMID- 9724614 TI - Myosin rod-packing schemes in vertebrate muscle thick filaments. AB - Muscle myosin filament backbones are known to be aggregates of long coiled-coil alpha-helical myosin rods, but the packing arrangement is not understood in detail. Here we present new data on fish muscle myosin filaments from low-angle X ray diffraction and from freeze-fracture, deep-etch electron microscopy which put constraints on the kind of models that might explain all of the observations. In particular, it is known in the case of vertebrate striated muscle thick filaments that the myosin head array in resting muscle is not perfectly helical but contains periodic perturbations. We show by analysis of low-angle X-ray diffraction patterns from resting bony fish muscle that any radial, azimuthal, and axial perturbations of the myosin head origins on the filament surface (due to perturbed myosin rod packing) must all be rather small and that the main perturbations are in the myosin head configurations (i.e., tilts, slews, rotations) on those origins. We provide evidence that the likely arrangement of titin molecules on the myosin filament is with them aligned parallel to the filament long axis, rather than following helical tracks. We also show from freeze-fracture studies of fish muscle that the myosin filament backbone (including titin and other extra proteins) has a radius of about 65-75 A and appears to contain a small (approximately 15-20 A radius) hollow core. Together with previously published evidence showing that the myosin rods are nearly parallel to the thick filament long axis, these results are consistent with the curved crystalline layer model of Squire (J. M. Squire, 1973, J. Mol. Biol. 77, 291-323), and they suggest a general structure for the C-zone part of the thick filament PMID- 9724615 TI - Changes in interfilament spacing mimic the effects of myosin regulatory light chain phosphorylation in rabbit psoas fibers. AB - The modulatory effect of myosin regulatory light chain phosphorylation in mammalian skeletal muscle, first documented as posttetanic potentiation of twitch tension, was subsequently shown to enhance the expression and development of tension at submaximal levels of activating calcium. Structural analyses demonstrated that thick filaments with phosphorylated myosin regulatory light chains appeared disordered: they lost the near-helical, periodic arrangement of myosin head characteristic of the relaxed state. We suggested that disordered heads may be more mobile than ordered heads and are likely to spend more time close to their binding sites on thin filaments. In this study we determined that the physiological effects of phosphorylation could be mimicked by decreasing the lattice spacing between the thick and the thin filaments, either by osmotic compression with dextran or by increasing the sarcomere length of permeabilized rabbit psoas fibers. Phosphorylation of regulatory light chains by incubation of permeabilized fibers with myosin light chain kinase and calmodulin, followed by low levels of activating calcium, potentiated tension development at resting or lower sarcomere lengths in the absence of dextran but had no additional effect on tension potentiation or development in fibers with decreased lattice spacing due to either osmotic compression or increased sarcomere length. PMID- 9724616 TI - Structural and functional responses of mammalian thick filaments to alterations in myosin regulatory light chains. AB - The ordered array of myosin heads, characteristic of relaxed striated muscle thick filaments, is reversibly disordered by phosphorylating myosin regulatory light chains, decreasing temperature and/or ionic strength, increasing pH, and depleting nucleotide. In the case of light chain phosphorylation, disorder, most likely due to a change in charge affecting the light chain amino-terminus, reflects increased myosin head mobility, thus increased accessibility to actin, and results in increased calcium sensitivity of tension development. Thus, interactions between the unphosphorylated regulatory light chain and the filament backbone may help maintain the overall order of the relaxed filament. To define this relationship, we have examined the structural and functional effects of such manipulations as exchanging wild-type smooth and skeletal myosin light chains into permeabilized rabbit psoas fibers and removing regulatory light chains (without exchange) from such fibers. We have also compared the structural and functional parameters of biopsied fibers from patients with severe familial hypertrophic cardiomyopathy due to a single amino acid substitution in the regulatory light chains to those exhibited by fibers from normal relatives. Our results support a role for regulatory light chains in reversible ordering of myosin heads and suggest that economy of energy utilization may provide for evolutionary preservation of this function in vertebrate striated muscle. PMID- 9724617 TI - Differential protein localization in sarcomeric and nonsarcomeric contractile structures of cultured cardiomyocytes. AB - The use of cardiomyocyte cell culture models allows the identification of various cell mediators that bring about changes in subcellular structures and gene expression associated with hypertrophy. The effects of insulin-like growth factor I (IGF-I), basic fibroblast growth factor (bFGF), and triiodothyronine (T3) on gene expression and on the structural organization of myofibrillar and cytoskeletal proteins were compared in adult atrial (aARC) and ventricular (vARC) as well as in neonatal ventricular rat cardiomyocytes (vNRC) in long-term culture. Structural changes were evaluated by confocal microscopy and correlated to biochemical alterations. In vARC, IGF-I enhanced myofibrillar growth, whereas bFGF or T3 restricted sarcomere assembly to the central cell area, forming a sharp boundary in more than 50% of the cells. However, myosin occurred both in the cross-striated myofibrillar structures and in patches running along the nonsarcomeric fibrillar structures (also called stress fiber-like structures) in the cell periphery. In cells treated with either bFGF or T3, the expression of alpha-smooth muscle actin (alpha-sm actin) was greatly increased. This actin isoform was incorporated mainly into the nonsarcomeric contractile structures outside the area where myofibrils ended abruptly. alpha-sm actin protein increased up to 14- to 17-fold while the mRNA showed a moderate increase of 2- to 4-fold. This suggests that alpha-sm actin is mainly regulated at the translational or posttranslational level. In contrast, the cytoskeletal proteins alpha-actinin and vinculin increased only moderately (less than 2-fold) but also showed a relocalization in cells with restricted myofibrils. In aARC and in vNRC, alpha-sm actin was only moderately upregulated by bFGF or T3 and no drastic morphological changes were observed. In conclusion, IGF-I, bFGF, and T3 induced characteristic structural phenotypes depending on the type of cardiomyocyte. Large amounts of alpha-sm actin as expressed in bFGF and T3 treated vARC seem to be incompatible with sarcomere assembly. PMID- 9724618 TI - The stability of tropomyosin at acid pH: effects of anion binding. AB - The alpha-helical coiled-coil tropomyosin homodimer, alpha alphaTm, unfolds cooperatively with T1/2 = 47 degreesC, at neutral pH, 0.5 M NaCl. At pH 2, where each chain contains 55 positive charges, no cooperative unfolding occurs to at least 80 degreesC. The NaCl and K2SO4 dependence of thermal unfolding of alpha alphaTm and a less stable protein, beta betaTm, were studied with circular dichroism. For alpha alphaTm, 10 mM NaCl, or 0.5 mM K2SO4, was sufficient to increase the unfolding temperature by 80 degreesC. For beta betaTm, similar concentrations of NaCl and K2SO4 as for alpha alphaTm increased the unfolding temperature of the most stable domain. Titrations indicated that two to three anions bind preferentially to the beta betaTm intermediate. Thus anions bind to Tm at acid pH values to greatly stabilize the helix. But even in the absence of added salt, Tm is more stable at pH 2 than pH 7, suggesting destabilization by negatively charged amino acids. PMID- 9724619 TI - A conserved C-terminal assembly region in paramyosin and myosin rods. AB - The assembly of myosin and paramyosin into filaments in muscle has been shown to depend in part on the interactions of regular periodic patches of charge on the surface of the rod regions of these alpha-helical coiled-coil proteins. It has also been known for some time that a relatively small region near the C-terminus of both molecules is critical for both solubility and assembly. This domain appears to function as a modulator of assembly in both proteins. Recently, a specific 29-residue region in the C-terminus of human fast muscle myosin rod has been shown to be essential for filament formation, and this sequence has been shown to be present in other vertebrate and invertebrate myosins. We show here that paramyosin also displays this specific conserved domain. Moreover, we have found that this domain is part of a longer distinctive region in both paramyosin and myosin: this region lacks the periodic variation in charge found in the rest of both coiled coils, has a unique charge profile, a relatively neutral total charge, and a high proportion of large apolar residues in surface positions. These results may be useful in designing site-directed mutagenesis studies to identify the target regions on neighboring molecules which interact with this C terminal domain and so establish the mechanism of its function. PMID- 9724621 TI - Complete unfolding of the titin molecule under external force. AB - Titin (also known as connectin) is a giant filamentous protein that spans the distance between the Z- and M-lines of the vertebrate muscle sarcomere. Several earlier studies have implicated titin as playing a fundamental role in maintaining sarcomeric structural integrity and generating the passive force of muscle. The elastic properties of titin were characterized in recent single molecule mechanical works that described the molecule as an entropic spring in which partial unfolding may take place at high forces during stretch and refolding at low forces during release. In the present work titin molecules were stretched using a laser tweezer with forces above 400 pN. The high external forces resulted in complete mechanical unfolding of the molecule, characterized by the disappearance of force hysteresis at high forces. Titin refolded following complete denaturation, as the hysteresis at low forces reappeared in subsequent stretch-release cycles. The broad force range throughout which unfolding occurred indicates that the various globular domains in titin require different unfolding forces due to differences in the activation energies for their unfolding. PMID- 9724620 TI - PEVK extension of human soleus muscle titin revealed by immunolabeling with the anti-titin antibody 9D10. AB - Titin is a giant protein that spans half of the striated muscle sarcomere. The I band portion of titin extends as the sarcomere is stretched, developing what is known as passive force. This portion of the molecule is composed mainly of tandem immunoglobulin (Ig) segments, consisting of serially linked Ig-like domains, and a recently discovered unique sequence termed the PEVK segment. The tandem Ig and PEVK segments have been suggested to extend sequentially when sarcomeres are stretched, with PEVK extension dominating at moderate to high degrees of sarcomere stretch (M. Gautel and D. Goulding, 1996, FEBS Lett. 385, 11-14; W. Linke et al., 1996, J. Mol. Biol. 261, 62-71; K. Trombitas et al., 1998). Previously we observed that the anti-titin antibody 9D10 labels a region in the I band that increases in width as sarcomeres are stretched. Here we tested whether 9D10 labels the PEVK segment. The 9D10-labeled region of human soleus fibers was followed by immunoelectron microscopy as sarcomeres were stretched. It was found that 9D10 labeled a region in the I-band that was approximately 100 nm wide at a sarcomere length of 2.4 micrometer and approximately 550 nm wide at a sarcomere length of 4.0 micrometer. Results were compared with those obtained with sequence specific antibodies that were used to mark the boundaries of the PEVK segment. Findings indicate that 9D10 labels the PEVK segment from close to its N-terminal end to its C-terminal end. Consistent with this conclusion are the results on cardiac myocytes that express a much shorter PEVK segment than skeletal muscle and where 9D10 labels a region that is much less wide than in skeletal muscle. The anti-titin antibody 9D10 is a useful tool for investigating the extensible behavior of the PEVK segment in both skeletal and cardiac muscles. PMID- 9724622 TI - A survey of the primary structure and the interspecies conservation of I-band titin's elastic elements in vertebrates. AB - Titin is a >3000-kDa large filamentous protein of vertebrate-striated muscle, and single titin molecules extend from the Z disc to the M line. In its I-band section, titin behaves extensible and is responsible for myofibrillar passive tension during stretch. However, details of the molecular basis of titin's elasticity are not known. We have compared the motif sequences of titin elastic elements from different vertebrate species and from different regions of the molecule. The I-band titin Ig repeats that are expressed in the stiff cardiac muscle and those that are tissue-specifically expressed in more elastic skeletal muscles represent distinct subgroups. Within the tissue-specifically expressed Ig repeats, a super-repeat structure is found. For the PEVK titin sequences, we surveyed interspecies conservation by hybridization experiments. The sequences of the titin gene which code for the C-terminal region of the PEVK domain are conserved in the genomes of a larger variety of vertebrates, whereas the N terminal PEVK sequences are more divergent. Future comparisons of titin gene sequences from different vertebrates may improve our understanding of how titin contributes to species diversity of myofibrillar elasticity. Within one species, different classes of Ig repeat families may contribute to elastic diversity of the titin spring in different segments. PMID- 9724623 TI - Folding and function of repetitive structure in the homotrimeric phage P22 tailspike protein. AB - The Salmonella bacteriophage P22 recognizes its host cell receptor, lipopolysaccharide, by means of six tailspikes, thermostable homotrimers of 72 kDa polypeptides. Biophysical results on the binding reaction, together with high resolution structural information from X-ray crystallography, have shed light on the interactions determining the viral host range. Folding and assembly of the tailspike protein in vitro have been analyzed in detail, and the data have been compared with observations on the in vivo assembly pathway. Repetitive structural elements in the tailspike protein, like a side-by-side trimer of parallel beta helices, a parallel alpha-helical bundle, a triangular prism made up from antiparallel beta-sheets, and a short segment of a triple beta-helix can be considered building blocks for larger structural proteins, and thus, the results on P22 tailspike may have implications for fibrous protein structure and folding. PMID- 9724624 TI - Sequence profile of the parallel beta helix in the pectate lyase superfamily. AB - The parallel beta helix structure found in the pectate lyase superfamily has been analyzed in detail. A comparative analysis of known structures has revealed a unique sequence profile, with a strong positional preference for specific amino acids oriented toward the interior of the parallel beta helix. Using the unique sequence profile, search patterns have been constructed and applied to the sequence databases to identify a subset of proteins that are likely to fold into the parallel beta helix. Of the 19 families identified, 39% are known to be carbohydrate-binding proteins, and 50% belong to a broad category of proteins with sequences containing leucine-rich repeats (LRRs). The most striking result is the sequence match between the search pattern and four contiguous segments of internalin A, a surface protein from the bacterial pathogen Listeria monocytogenes. A plausible model of the repetitive LRR sequences of internalin A has been constructed and favorable 3D-1D profile scores have been calculated. Moreover, spectroscopic features characteristic of the parallel beta helix topology in the pectate lyases are present in the circular dichroic spectrum of internalin A. Altogether, the data support the hypothesis that sequence search patterns can be used to identify proteins, including a subset of LRR proteins, that are likely to fold into the parallel beta helix. PMID- 9724625 TI - Structure and evolution of parallel beta-helix proteins. AB - Three bacterial pectate lyases, a pectin lyase from Aspergillus niger, the structures of rhamnogalacturonase A from Aspergillus aculeatus, RGase A, and the P22-phage tailspike protein, TSP, display the right-handed parallel beta-helix architecture first seen in pectate lyase. The lyases have 7 complete coils while RGase A and TSP have 11 and 12, respectively. Each coil contains three beta strands and three turn regions named PB1, T1, PB2, T2, PB3, and T3 in their order of occurrence. The lyases have homologous sequences but RGase A and TSP do not show obvious sequence homology either to the lyases or to each other. However, the structural similarities between all these molecules are so extensive that divergence from a common ancestor is much more probable than convergence to the same fold. The region PB2-T2-PB3 is the best conserved region in the lyases and shows the clearest structural similarity. Not only is the pleating and the direction of the hydrogen bonding in the sheets conserved, but so is the unusual alphaL-conformation turn between the two sheets. However, the overall shape, the position of long loops, a conserved alpha-helix that covers the amino-terminal end of the parallel beta-helix and stacks of residues in alphaR-conformation at the start of PB1 all suggest a common ancestor. The functional similarity, that the enzymes all bind alpha-galactose containing polymers at an equivalent site involving PB1 and its two flanking turn regions, further supports divergent evolution. We suggest that the stacking of the coils and the unusual near perpendicular junction of PB2 and PB3 make the parallel beta-helix fold especially likely to maintain similar main chain conformations during divergent evolution even after all vestige of similarity in primary structure has vanished. PMID- 9724626 TI - Polypeptide chain folding in the hydrophobic core of hamster scrapie prion: analysis by X-ray diffraction. AB - Conversion of the noninfectious, cellular form of the scrapie prion (PrPC) to the infectious form (PrPSc) is thought to be driven by an alpha-helical to beta-sheet conformational transition. The N-truncated polypeptide PrP27-30, which encompasses residues 90-231 of PrPSc and from which the truncated peptide is derived by limited proteolysis, assembles into amyloid rods that are rich in the beta-sheet conformation. The N-terminal half of PrP27-30, which includes residues 90-145 of PrP (SHa90-145) and contains the two putative alpha-helical domains H1 (PrP109-122) and H2 (PrP129-141), appears to be particularly crucial in the alpha --> beta conversion. To assess their role in this conformational transition, we have analyzed in detail X-ray diffraction patterns from the prion-related peptides A8A (PrP113-120), H1, and SHa90-145. We used iterative Fourier synthesis with beta-silk as an initial model for assigning phases. For H1, the lyophilized and acetonitrile-solubilized/dehydrated specimens gave two different electron density maps. The former showed that the beta-sheets were composed of small side chains as in A8A. The latter showed two types of beta-sheets having smaller and larger side chains, suggesting a turn. Such a turn was not observed in the lyophilized H1, indicating that the internal turn in H1 depends on the physical chemical environment. In SHa90-145, the beta-chains are assembled in approximately 40 A-wide crystal domains (termed beta-crystallites), and the electron density maps of these crystallites showed evidence for turns within both the H1 and H2 domains. The molecular folding of H1-H2 is compared here with the recent NMR solution structure of recombinant hamster prion, and the effect of pH on the conformational change is discussed. The most compact structure based on the X-ray diffraction analysis showed that the N-terminal, smaller residues of H2 fold back and are hydrogen-bonded with the C-terminal, smaller residues of H1. Similar folding is observed in the NMR solution structure. Comparison of the NMR structures at different pH with the X-ray diffraction results suggests that histidine and lysine residues in the N-terminal sequence of PrP may figure in the alpha --> beta structure transition of PrP. PMID- 9724627 TI - CpG methylation, chromatin structure and gene silencing-a three-way connection. AB - The three-way connection between DNA methylation, gene activity and chromatin structure has been known for almost two decades. Nevertheless, the molecular link between methyl groups on the DNA and the positioning of nucleosomes to form an inactive chromatin configuration was missing. This review discusses recent experimental data that may, for the first time, shed light on this molecular link. MeCP2, which is a known methylcytosine-binding protein, has been shown to possess a transcriptional repressor domain (TRD) that binds the corepressor mSin3A. This corepressor protein constitutes the core of a multiprotein complex that includes histone deacetylases (HDAC1 and HDAC2). Transfection and injection experiments with methylated constructs have revealed that the silenced state of a methylated gene, which is associated with a deacetylated nucleosomal structure, could be relieved by the deacetylase inhibitor, trichostatin A. Thus, methylation plays a pivotal role in establishing and maintaining an inactive state of a gene by rendering the chromatin structure inaccessible to the transcription machinery. PMID- 9724628 TI - Active site mutants in the six regulatory particle ATPases reveal multiple roles for ATP in the proteasome. AB - A family of ATPases resides within the regulatory particle of the proteasome. These proteins (Rpt1-Rpt6) have been proposed to mediate substrate unfolding, which may be required for translocation of substrates through the channel that leads from the regulatory particle into the proteolytic core particle. To analyze the role of ATP hydrolysis in protein breakdown at the level of the individual ATPase, we have introduced equivalent site-directed mutations into the ATPbinding motif of each RPT gene. Non-conservative substitutions of the active-site lysine were lethal in four of six cases, and conferred a strong growth defect in two cases. Thus, the ATPases are not functionally redundant, despite their multiplicity and sequence similarity. Degradation of a specific substrate can be inhibited by ATP-binding-site substitutions in many of the Rpt proteins, indicating that they co-operate in the degradation of individual substrates. The phenotypic defects of the different rpt mutants were strikingly varied. The most divergent phenotype was that of the rpt1 mutant, which was strongly growth defective despite showing no general defect in protein turnover. In addition, rpt1 was unique among the rpt mutants in displaying a G1 cell-cycle defect. Proteasomes purified from an rpt2 mutant showed a dramatic inhibition of peptidase activity, suggesting a defect in gating of the proteasome channel. In summary, ATP promotes protein breakdown by the proteasome through multiple mechanisms, as reflected by the diverse phenotypes of the rpt mutants. PMID- 9724629 TI - DPM2 regulates biosynthesis of dolichol phosphate-mannose in mammalian cells: correct subcellular localization and stabilization of DPM1, and binding of dolichol phosphate. AB - Biosynthesis of glycosylphosphatidylinositol and N-glycan precursor is dependent upon a mannosyl donor, dolichol phosphate-mannose (DPM). The Thy-1negative class E mutant of mouse lymphoma and Lec15 mutant Chinese hamster ovary (CHO) cells are incapable of DPM synthesis. The class E mutant is defective in the DPM1 gene which encodes a mammalian homologue of Saccharomyces cerevisiae Dpm1p that is a DPM synthase, whereas Lec15 is a different mutant, indicating that mammalian DPM1 is not sufficient for DPM synthesis. Here we report expression cloning of a new gene, DPM2, which is defective in Lec15 cells. DPM2, an 84 amino acid membrane protein expressed in the endoplasmic reticulum (ER), makes a complex with DPM1 that is essential for the ER localization and stable expression of DPM1. Moreover, DPM2 enhances binding of dolichol phosphate, a substrate of DPM synthase. Mammalian DPM1 is catalytic because a fusion protein of DPM1 that was stably expressed in the ER synthesized DPM without DPM2. Therefore, biosynthesis of DPM in mammalian cells is regulated by DPM2. PMID- 9724630 TI - Phosphoinositide signaling and turnover: PtdIns(3)P, a regulator of membrane traffic, is transported to the vacuole and degraded by a process that requires lumenal vacuolar hydrolase activities. AB - The Golgi/endosome-associated Vps34 phosphatidylinositol 3-kinase is essential for the sorting of hydrolases from the Golgi to the vacuole/lysosome. Upon inactivation of a temperature-conditional Vps34 kinase, cellular levels of PtdIns(3)P rapidly decrease and it has been proposed that this decrease is due to the continued turnover of PtdIns(3)P by cytoplasmic phosphatases. Here we show that mutations in VAM3 (vacuolar t-SNARE) and YPT7 (rab GTPase), which are required to direct protein and membrane delivery from prevacuolar endosomal compartments to the vacuole, dramatically increase/stabilize PtdIns(3)P levels in vivo by disrupting its turnover. We find that the majority of the total pool of PtdIns(3)P which has been synthesized, but not PtdIns(4)P, requires transport to the vacuole in order to be turned over. Unexpectedly, strains with impaired vacuolar hydrolase activity accumulate 4- to 5-fold higher PtdIns(3)P levels than wild-type cells, suggesting that lumenal vacuolar lipase and/or phosphatase activities degrade PtdIns(3)P. Because vacuolar hydrolases act in the lumen, PtdIns(3)P is likely to be transferred from the cytoplasmic membrane leaflet where it is synthesized, to the lumen of the vacuole. Interestingly, mutants that stabilize PtdIns(3)P accumulate small uniformly-sized vesicles (40-50 nm) within prevacuolar endosomes (multivesicular bodies) or the vacuole lumen. Based on these and other observations, we propose that PtdIns(3)P is degraded by an unexpected mechanism which involves the sorting of PtdIns(3)P into vesicles generated by invagination of the limiting membrane of the endosome or vacuole, ultimately delivering the phosphoinositide into the lumen of the compartment where it can be degraded by the resident hydrolases. PMID- 9724631 TI - Clathrin coats at 21 A resolution: a cellular assembly designed to recycle multiple membrane receptors. AB - We present a map at 21 A resolution of clathrin assembled into cages with the endocytic adaptor complex, AP-2. The map was obtained by cryo-electron microscopy and single-particle reconstruction. It reveals details of the packing of entire clathrin molecules as they interact to form a cage with two nested polyhedral layers. The proximal domains of each triskelion leg depart from a cage vertex in a skewed orientation, forming a slightly twisted bundle with three other leg domains. Thus, each triskelion contributes to two connecting edges of the polyhedral cage. The clathrin heavy chains continue inwards under the vertices with local 3-fold symmetry, the terminal domains contributing to 'hook-like' features which form an intermediate network making possible contacts with the surface presented by the inner adaptor shell. A node of density projecting inwards from the vertex may correspond to the C-termini of clathrin heavy chains which form a protrusion on free triskelions at the vertex. The inter-subunit interactions visible in this map provide a structural basis for considering the assembly of clathrin coats on a membrane and show the contacts which will need to be disrupted during disassembly. PMID- 9724632 TI - Specific binding to a novel and essential Golgi membrane protein (Yip1p) functionally links the transport GTPases Ypt1p and Ypt31p. AB - The regulation of vesicular transport in eukaryotic cells involves Ras-like GTPases of the Ypt/Rab family. Studies in yeast and mammalian cells indicate that individual family members act in vesicle docking/fusion to specific target membranes. Using the two-hybrid system, we have now identified a 248 amino acid, integral membrane protein, termed Yip1, that specifically binds to the transport GTPases Ypt1p and Ypt31p. Evidence for physical interaction of these GTPases with Yip1p was also demonstrated by affinity chromatography and/or co immunoprecipitation. Like the two GTPases, Yip1p is essential for yeast cell viability and, according to subcellular fractionation and indirect immunofluorescence, is located to Golgi membranes at steady state. Mutant cells depleted of Yip1p and conditionally lethal yip1 mutants at the non-permissive temperature massively accumulate endoplasmic reticulum membranes and display aberrations in protein secretion and glycosylation of secreted invertase. The results suggests for a role for Yip1p in recruiting the two GTPases to Golgi target membranes in preparation for fusion. PMID- 9724633 TI - Muskelin, a novel intracellular mediator of cell adhesive and cytoskeletal responses to thrombospondin-1. AB - We have used an expression cloning strategy based on a cell-attachment assay screen to seek identification of molecules required in cellular responses to thrombospondin-1, a regulated macromolecular component of extracellular matrix. We report the identification and functional characterization of a novel, widely expressed, intracellular protein, named muskelin, which contains dispersed motifs with homology to the tandem repeats first identified in the Drosophila kelch ORF1 protein. In adherent C2C12 cells, muskelin localizes in the cytoplasm and at cell margins. Over-expression of muskelin in C2C12 cells promotes cell attachment to the thrombospondin-1 C-terminal domain, alters the mechanisms of attachment to intact thrombospondin-1 and correlates with decreased formation of fascin microspikes and increased assembly of focal contacts by cells adherent on thrombospondin-1. Reciprocally, cell attachment, spreading and cytoskeletal organization are specifically reduced in TSP-1-adherent cells after antisense depletion of muskelin. These results establish a requirement for muskelin in cell responses to thrombospondin-1 and demonstrate that such responses involve a novel process which is integrated into the regulation of cell-adhesive behaviour and cytoskeletal organization. PMID- 9724634 TI - Oligopeptidase B-dependent signaling mediates host cell invasion by Trypanosoma cruzi. AB - Mammalian cell invasion by the intracellular protozoan parasite Trypanosoma cruzi is mediated by recruitment and fusion of host cell lysosomes, an unusual process that has been proposed to be dependent on the ability of parasites to trigger intracellular free calcium concentration ([Ca2+]i) transients in host cells. Previous work implicated the T.cruzi serine hydrolase oligopeptidase B in the generation of Ca2+-signaling activity in parasite extracts. Here we show that deletion of the gene encoding oligopeptidase B results in a marked defect in host cell invasion and in the establishment of infections in mice. The invasion defect is associated with the inability of oligopeptidase B null mutant trypomastigotes to mobilize Ca2+ from thapsigargin-sensitive stores in mammalian cells. Exogenous recombinant oligopeptidase B reconstitutes the oligopeptidase B-dependent Ca2+ signaling activity in null mutant parasite extracts, demonstrating that this enzyme is responsible for the generation of a signaling agonist for mammalian cells. PMID- 9724635 TI - Integrating cytosolic calcium signals into mitochondrial metabolic responses. AB - Stimulation of hepatocytes with vasopressin evokes increases in cytosolic free Ca2+ ([Ca2+]c) that are relayed into the mitochondria, where the resulting mitochondrial Ca2+ ([Ca2+]m) increase regulates intramitochondrial Ca2+-sensitive targets. To understand how mitochondria integrate the [Ca2+]c signals into a final metabolic response, we stimulated hepatocytes with high vasopressin doses that generate a sustained increase in [Ca2+]c. This elicited a synchronous, single spike of [Ca2+]m and consequent NAD(P)H formation, which could be related to changes in the activity state of pyruvate dehydrogenase (PDH) measured in parallel. The vasopressin-induced [Ca2+]m spike evoked a transient increase in NAD(P)H that persisted longer than the [Ca2+]m increase. In contrast, PDH activity increased biphasically, with an initial rapid phase accompanying the rise in [Ca2+]m, followed by a sustained secondary activation phase associated with a decline in cellular ATP. The decline of NAD(P)H in the face of elevated PDH activity occurred as a result of respiratory chain activation, which was also manifest in a calcium-dependent increase in the membrane potential and pH gradient components of the proton motive force (PMF). This is the first direct demonstration that Ca2+-mobilizing hormones increase the PMF in intact cells. Thus, Ca2+ plays an important role in signal transduction from cytosol to mitochondria, with a single [Ca2+]m spike evoking a complex series of changes to activate mitochondrial oxidative metabolism. PMID- 9724636 TI - The alternative product from the human CDKN2A locus, p14(ARF), participates in a regulatory feedback loop with p53 and MDM2. AB - The two distinct proteins encoded by the CDKN2A locus are specified by translating the common second exon in alternative reading frames. The product of the alpha transcript, p16(INK4a), is a recognized tumour suppressor that induces a G1 cell cycle arrest by inhibiting the phosphorylation of the retinoblastoma protein by the cyclin-dependent kinases, CDK4 and CDK6. In contrast, the product of the human CDKN2A beta transcript, p14(ARF), activates a p53 response manifest in elevated levels of MDM2 and p21(CIP1) and cell cycle arrest in both G1 and G2/M. As a consequence, p14(ARF)-induced cell cycle arrest is p53 dependent and can be abrogated by the co-expression of human papilloma virus E6 protein. p14(ARF) acts by binding directly to MDM2, resulting in the stabilization of both p53 and MDM2. Conversely, p53 negatively regulates p14(ARF) expression and there is an inverse correlation between p14(ARF) expression and p53 function in human tumour cell lines. However, p14(ARF) expression is not involved in the response to DNA damage. These results place p14(ARF) in an independent pathway upstream of p53 and imply that CDKN2A encodes two proteins that are involved in tumour suppression. PMID- 9724637 TI - A novel p53-inducible gene coding for a microtubule-localized protein with G2 phase-specific expression. AB - Wild-type (wt) p53 can act as a sequence-specific transcriptional activator and it is believed that p53 elicits at least part of its biological effects by regulating the expression of specific target genes. By using a differential subtractive hybridization approach in a murine cell line stably transfected with a temperature-sensitive p53 mutant (Val135), we isolated a set of genes markedly induced by wt p53. One of them, provisionally named B99, was further characterized; its transcriptional induction was dependent on wt p53 function and the corresponding protein product was shown to accumulate after DNA damage in different cell types. Immunofluorescence analysis located the B99 protein to the microtubule network. Flow cytometry revealed that upon activation of p53 function the endogenous B99 protein was selectively induced in the G2 fraction of the cell population. When B99 was ectopically expressed in p53-null murine fibroblasts, B99-transfected cells displayed an increased fraction with a 4N DNA content, indicative of interference with G2 phase progression. Taken together these data suggest that B99 might play a role in mediating specific biological activities of wt p53 during the G2 phase. PMID- 9724638 TI - Cell-cycle arrest and inhibition of G1 cyclin translation by iron in AFT1-1(up) yeast. AB - Although iron is an essential nutrient, it is also a potent cellular toxin, and the acquisition of iron is a highly regulated process in eukaryotes. In yeast, iron uptake is homeostatically regulated by the transcription factor encoded by AFT1. Expression of AFT1-1(up), a dominant mutant allele, results in inappropriately high rates of iron uptake, and AFT1-1(up) mutants grow slowly in the presence of high concentrations of iron. We present evidence that when Aft1 1(up) mutants are exposed to iron, they arrest the cell division cycle at the G1 regulatory point Start. This arrest is dependent on high-affinity iron uptake and does not require the activation of the DNA damage checkpoint governed by RAD9. The iron-induced arrest is bypassed by overexpression of a mutant G1 cyclin, cln3 2, and expression of the G1-specific cyclins Cln1 and Cln2 is reduced when yeast are exposed to increasing amounts of iron, which may account for the arrest. This reduction is not due to changes in transcription of CLN1 or CLN2, nor is it due to accelerated degradation of the protein. Instead, this reduction occurs at the level of Cln2 translation, a recently recognized locus of cell-cycle control in yeast. PMID- 9724639 TI - A link between MAP kinase and p34(cdc2)/cyclin B during oocyte maturation: p90(rsk) phosphorylates and inactivates the p34(cdc2) inhibitory kinase Myt1. AB - M-phase entry in eukaryotic cells is driven by activation of MPF, a regulatory factor composed of cyclin B and the protein kinase p34(cdc2). In G2-arrested Xenopus oocytes, there is a stock of p34(cdc2)/cyclin B complexes (pre-MPF) which is maintained in an inactive state by p34(cdc2) phosphorylation on Thr14 and Tyr15. This suggests an important role for the p34(cdc2) inhibitory kinase(s) such as Wee1 and Myt1 in regulating the G2-->M transition during oocyte maturation. MAP kinase (MAPK) activation is required for M-phase entry in Xenopus oocytes, but its precise contribution to the activation of pre-MPF is unknown. Here we show that the C-terminal regulatory domain of Myt1 specifically binds to p90(rsk), a protein kinase that can be phosphorylated and activated by MAPK. p90(rsk) in turn phosphorylates the C-terminus of Myt1 and down-regulates its inhibitory activity on p34(cdc2)/cyclin B in vitro. Consistent with these results, Myt1 becomes phosphorylated during oocyte maturation, and activation of the MAPK-p90(rsk) cascade can trigger some Myt1 phosphorylation prior to pre-MPF activation. We found that Myt1 preferentially associates with hyperphosphorylated p90(rsk), and complexes can be detected in immunoprecipitates from mature oocytes. Our results suggest that during oocyte maturation MAPK activates p90(rsk) and that p90(rsk) in turn down-regulates Myt1, leading to the activation of p34(cdc2)/cyclin B. PMID- 9724640 TI - Cysteine string protein (CSP) is an insulin secretory granule-associated protein regulating beta-cell exocytosis. AB - Cysteine string proteins (CSPs) are novel synaptic vesicle-associated protein components characterized by an N-terminal J-domain and a central palmitoylated string of cysteine residues. The cellular localization and functional role of CSP was studied in pancreatic endocrine cells. In situ hybridization and RT-PCR analysis demonstrated CSP mRNA expression in insulin-producing cells. CSP1 mRNA was present in pancreatic islets; both CSP1 and CSP2 mRNAs were seen in insulin secreting cell lines. Punctate CSP-like immunoreactivity (CSP-LI) was demonstrated in most islets of Langerhans cells, acinar cells and nerve fibers of the rat pancreas. Ultrastructural analysis showed CSP-LI in close association with membranes of secretory granules of cells in the endo- and exocrine pancreas. Subcellular fractionation of insulinoma cells showed CSP1 (34/36 kDa) in granular fractions; the membrane and cytosol fractions contained predominantly CSP2 (27 kDa). The fractions also contained proteins of 72 and 70 kDa, presumably CSP dimers. CSP1 overexpression in INS-1 cells or intracellular administration of CSP antibodies into mouse ob/ob beta-cells did not affect voltage-dependent Ca2+ channel activity. Amperometric measurements showed a significant decrease in insulin exocytosis in individual INS-1 cells after CSP1 overexpression. We conclude that CSP is associated with insulin secretory granules and that CSP participates in the molecular regulation of insulin exocytosis by mechanisms not involving changes in the activity of voltage-gated Ca2+-channels. PMID- 9724641 TI - G protein hyperactivation of the Caenorhabditis elegans adenylyl cyclase SGS-1 induces neuronal degeneration. AB - Expression of a constitutively activated version of the heterotrimeric G protein alpha-subunit Galphas results in the swelling and vacuolization of a specific subset of ventral nerve cord motoneurons of Caenorhabditis elegans. A second site modifier (sgs-1) that completely suppresses this neuronal degeneration has been isolated. sgs-1 was cloned and was shown to encode an adenylyl cyclase which is most similar to mammalian adenylyl cyclase type IX. Mutations in sgs-1 change residues that are conserved among different adenylyl cyclases. These mutations are located in the two catalytic domains and in the first multiple transmembrane spanning region of the predicted protein. An sgs-1 reporter construct shows a general neuronal expression pattern, demonstrating that sgs-1 is expressed in the neurons that are susceptible to activated Galphas-induced cell death. A second C.elegans adenylyl cyclase gene (acy-2) was analyzed as well. In contrast to sgs 1, acy-2 shows a restricted expression pattern and loss of acy-2 function results in early larval lethality. These results suggest that SGS-1 is a target of Galphas signaling in motoneurons, whereas an interaction of Galphas with ACY-2, probably in the canal-associated neurons, is required for viability. PMID- 9724642 TI - OCA-B integrates B cell antigen receptor-, CD40L- and IL 4-mediated signals for the germinal center pathway of B cell development. AB - Many of the key decisions in lymphocyte differentiation and activation are dependent on integration of antigen receptor and co-receptor signals. Although there is significant understanding of these receptors and their signaling pathways, little is known about the molecular requirements for signal integration at the level of activation of gene expression. Here we show that in primary B cells, expression of the B-cell specific transcription coactivator OCA-B (also known as OBF-1 or Bob-1) is regulated synergistically by the B-cell antigen receptor, CD40L and interleukin signaling pathways. Consistent with the requirement for multiple T cell-dependent signals to induce OCA-B, we find that OCA-B protein is highly expressed in germinal center B cells. Accordingly, germinal center formation is blocked completely in the absence of OCA-B expression in B cells, whereas the helper functions of OCA-B-deficient T cells are indistinguishable from controls. The requirement for OCA-B expression in B cells is germinal center specific since the development of primary B cell follicles, the marginal zone and plasma cells are all intact. Thus, OCA-B is the first example of a transcriptional coactivator that is both synergistically induced by and required for integration of signals that mediate cell fate decisions. PMID- 9724643 TI - Temperature-sensitive Gbeta mutants discriminate between G protein-dependent and independent signaling mediated by serpentine receptors. AB - Deletion of the single gene for the Dictyostelium G protein beta-subunit blocks development at an early stage. We have now isolated temperature-sensitive alleles of Gbeta to investigate its role in later development. We show that Gbeta is directly required for adenylyl cyclase A activation and for morphogenetic signaling during the entire developmental program. Gbeta was also essential for induction of aggregative gene expression by cAMP pulses, a process that is mediated by serpentine cAMP receptors (cARs). However, Gbeta was not required for cAR-mediated induction of prespore genes and repression of stalk genes, and neither was Gbeta needed for induction of prestalk genes by the differentiation inducing factor (DIF). cAMP induction of prespore genes and repression of stalk genes is mediated by the protein kinase GSK-3. GSK-3 also determines cell-type specification in insects and vertebrates and is regulated by the wingless/wnt morphogens that are detected by serpentine fz receptors. The G protein-dependent and -independent modes of cAR-mediated signaling reported here may also exist for the wingless/wnt signaling pathways in higher organisms. PMID- 9724644 TI - Insulin induces transcription of target genes through the hypoxia-inducible factor HIF-1alpha/ARNT. AB - Hypoxic stress induces the expression of genes associated with increased energy flux, including the glucose transporters Glut1 and Glut3, several glycolytic enzymes, nitric oxide synthase, tyrosine hydroxylase, erythropoietin and vascular endothelial growth factor (VEGF). Induction of these genes is mediated by a common basic helix-loop-helix-PAS transcription complex, the hypoxia-inducible factor-1alpha (HIF-1alpha)/aryl hydrocarbon nuclear translocator (ARNT). Insulin also induces some of these genes; however, the underlying mechanism is unestablished. We report here that insulin shares with hypoxia the ability to induce the HIF-1alpha/ARNT transcription complex in various cell types. This induction was demonstrated by electrophoretic mobility shift of the hypoxia response element (HRE), and abolished by specific antisera to HIF-1alpha and ARNT, and by transcription activation of HRE reporter vectors. Furthermore, basal and insulin-induced expression of Glut1, Glut3, aldolase A, phosphoglycerate kinase and VEGF was reduced in cells having a defective ARNT. Similarly, the insulin-induced activation of HRE reporter vectors and VEGF was impaired in these cells and was rescued by re-introduction of ARNT. Finally, insulin-like growth factor-I (IGF-I) also induced the HIF-1alpha/ARNT transcription complex. These observations establish a novel signal transduction pathway of insulin and IGF-I and broaden considerably the scope of activity of HIF-1alpha/ARNT. PMID- 9724645 TI - Transcription through a simple DNA repeat blocks replication elongation. AB - The influence of d(G)n.d(C)n repeats on plasmid replication in Escherichia coli cells was analyzed using electrophoretic analysis of replication intermediates. These repeats impeded the replication fork in a length- and orientation-dependent manner. Unexpectedly, the replication arrest relied primarily on the repeats' transcription. When the d(C)n sequence served as the transcriptional template, both transcription and replication were blocked. This was true for transcription driven by either bacterial or phage RNA polymerases. We hypothesize that the replication fork halts after it encounters a stalled ternary complex of the RNA polymerase, the DNA template and the r(G)n transcript. This constitutes a novel mechanism for the regulation of replication elongation. The effects of this mechanism on repeat length polymorphism and genome rearrangements are discussed. PMID- 9724647 TI - Conformational changes necessary for gene regulation by Tet repressor assayed by reversible disulfide bond formation. AB - We constructed and characterized four Tet repressor (TetR) variants with engineered cysteine residues which can form disulfide bonds and are located in regions where conformational changes during induction by tetracycline (tc) might occur. All TetR mutants show nearly wild-type activities in vivo, and the reduced proteins also show wild-type activities in vitro. Complete and reversible disulfide bond formation was achieved in vitro for all four mutants. The disulfide bond in NC18RC94 immobilizes the DNA reading head with respect to the protein core and prevents operator binding. Formation of this disulfide bond is possible only in the tc-bound, but not in the operator-bound conformation. Thus, these residues must have different conformations when bound to these ligands. The disulfide bonds in DC106PC159' and EC107NC165' immobilize the variable loop between alpha-helices 8 and 9 located near the tc-binding pocket. A faster rate of disulfide formation in the operator-bound conformation and a lack of induction after disulfide formation show that the variable loop is located closer to the protein core in the operator-bound conformation and that a movement is necessary for induction. The disulfide bond in RC195VC199' connects alpha-helices 10 and 10' of the two subunits in the dimer and is only formed in the tc-bound conformation. The oxidized protein shows reduced operator binding. Thus, this bond prevents formation of the operator-bound conformation. The detection of conformational changes in three different regions is the first biochemical evidence for induction-associated global internal movements in TetR. PMID- 9724646 TI - Interaction of PC4 with melted DNA inhibits transcription. AB - PC4 is a nuclear DNA-binding protein that stimulates activator-dependent class II gene transcription in vitro. Recent biochemical and X-ray analyses have revealed a unique structure within the C-terminal domain of PC4 that binds tightly to unpaired double-stranded (ds)DNA. The cellular function of this evolutionarily conserved dimeric DNA-binding fold is unknown. Here we demonstrate that PC4 represses transcription through this motif. Interaction with melted promoters is not required for activator-dependent transcription in vitro. The inhibitory activity is attenuated on bona fide promoters by (i) transcription factor TFIIH and (ii) phosphorylation of PC4. PC4 remains a potent inhibitor of transcription in regions containing unpaired ds DNA, in single-stranded DNA that can fold into two antiparallel strands, and on DNA ends. Our observations are consistent with a novel inhibitory function of PC4. PMID- 9724648 TI - Active recruitment of sigma54-RNA polymerase to the Pu promoter of Pseudomonas putida: role of IHF and alphaCTD. AB - The sequence elements determining the binding of the sigma54-containing RNA polymerase (sigma54-RNAP) to the Pu promoter of Pseudomonas putida have been examined. Contrary to previous results in related systems, we show that the integration host factor (IHF) binding stimulates the recruitment of the enzyme to the -12/-24 sequence motifs. Such a recruitment, which is fully independent of the activator of the system, XylR, requires the interaction of the C-terminal domain of the alpha subunit of RNAP with specific DNA sequences upstream of the IHF site which are reminiscent of the UP elements in sigma70 promoters. Our data show that this interaction is mainly brought about by the distinct geometry of the promoter region caused by IHF binding and the ensuing DNA bending. These results support the view that binding of sigma54-RNAP to a promoter is a step that can be subjected to regulation by factors (e.g. IHF) other than the sole intrinsic affinity of sigma54-RNAP for the -12/-24 site. PMID- 9724649 TI - Cloning and characterization of mCtBP2, a co-repressor that associates with basic Kruppel-like factor and other mammalian transcriptional regulators. AB - Basic Kruppel-like factor (BKLF) is a zinc finger protein that recognizes CACCC elements in DNA. It is expressed highly in erythroid tissues, the brain and other selected cell types. We have studied the activity of BKLF and found that it is capable of repressing transcription, and have mapped its repression domain to the N-terminus. We carried out a two-hybrid screen against BKLF and isolated a novel clone encoding murine C-terminal-binding protein 2 (mCtBP2). mCtBP2 is related to human CtBP, a cellular protein which binds to a Pro-X-Asp-Leu-Ser motif in the C terminus of the adenoviral oncoprotein, E1a. We show that mCtBP2 recognizes a related motif in the minimal repression domain of BKLF, and the integrity of this motif is required for repression activity. Moreover, when tethered to a promoter by a heterologous DNA-binding domain, mCtBP2 functions as a potent repressor. Finally, we demonstrate that mCtBP2 also interacts with the mammalian transcripition factors Evi-1, AREB6, ZEB and FOG. These results establish a new member of the CtBP family, mCtBP2, as a mammalian co-repressor targeting diverse transcriptional regulators. PMID- 9724650 TI - Binding of trithorax and Polycomb proteins to the bithorax complex: dynamic changes during early Drosophila embryogenesis. AB - In Drosophila, the maintenance of developmentally important transcription patterns is controlled at the level of chromatin structure. The Polycomb group (PcG) and trithorax group (trxG) genes encode proteins involved in chromatin remodelling. PcG genes have been proposed to act by packaging transcriptional repressed chromosomal domains into condensed heterochromatin-like structures. Some of the trxG proteins characterized so far are members of chromatin opening complexes (e.g. SWI/SNF and GAGA/NURF) which facilitate binding of transcription factors and components of the basal transcriptional machinery. Genetic and biochemical data suggest that these two groups of regulatory factors may act through a common set of DNA elements. In the present study, we have investigated the binding of Trithorax (TRX) and Polycomb (PC) protein in the bithorax complex (BX-C) during embryogenesis. In addition, we have identified the minimal fragments from the Ultrabithorax (Ubx) regulatory region that are capable of recruiting TRX to chromosomal sites containing them. Comparative analysis of the binding of the two proteins shows that TRX and PC bind target sequences (PcG regulated elements, PREs) by cellular blastoderm, when BX-C transcription begins. At the same stage, TRX but not PC is strongly associated with core promoters. Later, at germ band extension, the time of derepression in Polycomb mutants, PC binding is also detected outside core PREs and additionally binds to the fragments containing promoters. PMID- 9724651 TI - Chromatin immunoselection defines a TAL-1 target gene. AB - Despite the major functions of the basic helix-loop-helix transcription factor TAL-1 in hematopoiesis and T-cell leukemogenesis, no TAL-1 target gene has been identified. Using immunoprecipitation of genomic fragments bound to TAL-1 in the chromatin of murine erythro-leukemia (MEL) cells, we found that 10% of the immunoselected fragments contained a CAGATG or a CAGGTG E-box, followed by a GATA site. We studied one of these fragments containing two E-boxes, CAGATG and CAGGTC, followed by a GATA motif, and showed that TAL-1 binds to the CAGGTG E-box with an affinity modulated by the CAGATG or the GATA site, and that the CAGGTG GATA motif exhibits positive transcriptional activity in MEL but not in HeLa cells. This immunoselected sequence is located within an intron of a new gene co expressed with TAL-1 in endothelial and erythroid cells, but not expressed in fibroblasts or adult liver where no TAL-1 mRNA was detected. Finally, in vitro differentiation of embryonic stem cells towards the erythro/megakaryocytic pathways showed that the TAL-1 target gene expression followed TAL-1 and GATA-1 expression. These results establish that TAL-1 is likely to activate its target genes through a complex that binds an E-box-GATA motif and define the first gene regulated by TAL-1. PMID- 9724652 TI - TAFII105 mediates activation of anti-apoptotic genes by NF-kappaB. AB - The transcription factor NF-kappaB is important for expression of genes involved in immune responses, viral infections, cytokine signaling and stress. In addition NF-kappaB plays a crucial role in protecting cells from TNF-alpha-induced apoptotic stimuli, presumably by activating anti-apoptotic genes. Here we report that the sub-stoichiometric TFIID subunit TAFII105 is essential for activation of anti-apoptotic genes in response to TNF-alpha, serving as a transcriptional coactivator for NF-kappaB. The putative coactivator domain of TAFII105 interacts with the activation domain of the p65/RelA member of the NF-kappaB family, and further stimulates p65-induced transcription in human 293 cells. Moreover, inhibition of TAFII105 activity by overexpression of a dominant negative mutant of TAFII105 decreased NF-kappaB transcriptional activity and severely reduced cell survival in response to TNF-alpha. Similarly, expression of anti-sense TAFII105 RNA sensitized the cells to TNF-alpha cytotoxicity. These results suggest that TAFII105 is involved in activation of anti-apoptotic genes by NF kappaB. PMID- 9724653 TI - Sequential DNA damage-independent and -dependent activation of NF-kappaB by UV. AB - NF-kappaB activation in response to UV irradiation of HeLa cells or of primary human skin fibroblasts occurs with two overlapping kinetics but totally different mechanisms. Although both mechanisms involve induced dissociation of NF-kappaB from IkappaBalpha and degradation of IkappaBalpha, targeting for degradation and signaling are different. Early IkappaBalpha degradation at 30 min to approximately 6 h is not initiated by UV-induced DNA damage. It does not require IkappaB kinase (IKK), as shown by introduction of a dominant-negative kinase subunit, and does not depend on the presence of the phosphorylatable substrate, IkappaBalpha, carrying serines at positions 32 and 36. Induced IkappaBalpha degradation requires, however, intact N- (positions 1-36) and C-terminal (positions 277-287) sequences. IkappaB degradation and NF-kappaB activation at late time points, 15-20 h after UV irradiation, is mediated through DNA damage induced cleavage of IL-1alpha precursor, release of IL-1alpha and autocrine/paracrine action of IL-1alpha. Late-induced IkappaBalpha requires the presence of Ser32 and Ser36. The late mechanism indicates the existence of signal transfer from photoproducts in the nucleus to the cytoplasm. The release of the 'alarmone' IL-1alpha may account for some of the systemic effects of sunlight exposure. PMID- 9724654 TI - Association of RPA with chromosomal replication origins requires an Mcm protein, and is regulated by Rad53, and cyclin- and Dbf4-dependent kinases. AB - Eukaryotic cells use multiple replication origins to replicate their large genomes. Some origins fire early during S phase whereas others fire late. In Saccharomyces cerevisiae, initiator sequences (ARSs) are bound by the origin recognition complex (ORC). Cdc6p synthesized at the end of mitosis joins ORC and facilitates recruitment of Mcm proteins, which renders origins competent to fire. However, origins fire only upon the subsequent activation of S phase cyclin dependent kinases (S-CDKs) and Dbf4/Cdc7 at the G1/S boundary. We have used a chromatin immunoprecipitation assay to measure the association with ARS sequences of DNA primase and the single-stranded DNA binding replication protein A (RPA) when fork movement is inhibited by hydroxyurea (HU). RPA's association with origins requires S-CDKs, Dbf4/Cdc7 kinase and an Mcm protein. The recruitment of DNA primase depends on RPA. Furthermore, early- and late-firing origins differ not in the timing of their recruitment of an Mcm protein, but in the timing of RPA's recruitment. RPA is recruited to early but not to late origins in HU. We also show that Rad53 kinase is required to prevent RPA association with a late origin in HU. Our data suggest that the origin unwinding accompanied by RPA association is a key step, regulated by S-CDKs, Dbf4/Cdc7 and Rad53p. Thus, in the presence of active S-CDKs and Dbf4/Cdc7, Mcms may open origins and thereby facilitate the loading of RPA. PMID- 9724655 TI - Mechanistic studies of initiator-initiator interaction and replication initiation. AB - Unlike the chromosome of Escherichia coli that needs only one replication initiator protein (origin recognition protein) called DnaA, many plasmid replicons require dual initiators: host-encoded DnaA and a plasmid-encoded origin recognition protein, which is believed to be the major determinant of replication control. Hitherto, the relative mechanistic roles of dual initiators in DNA replication were unclear. Here, we present the first evidence that DnaA communicates with the plasmid-encoded pi initiator of R6K and contacts the latter at a specific N-terminal region. Without this specific contact, productive unwinding of plasmid ori gamma and replication is abrogated. The results also show that DnaA performs different roles in host and plasmid replication as revealed by the finding that the ATP-activated form of DnaA, while indispensable for oriC replication, was not required for R6K replication. We have analyzed the accessory role of the DNA bending protein, integration host factor (IHF), in promoting initiator-origin interaction and have found that IHF significantly enhances the binding of DnaA to its cognate site. Collectively, the results further advance our understanding of replication initiation. PMID- 9724656 TI - Copy number control of IncIalpha plasmid ColIb-P9 by competition between pseudoknot formation and antisense RNA binding at a specific RNA site. AB - Replication of a low-copy-number IncIalpha plasmid ColIb-P9 depends on expression of the repZ gene encoding the replication initiator protein. repZ expression is negatively controlled by the small antisense Inc RNA, and requires formation of a pseudoknot in the RepZ mRNA consisting of stem-loop I, the Inc RNA target, and a downstream sequence complementary to the loop I. The loop I sequence comprises 5' rUUGGCG-3', conserved in many prokaryotic antisense systems, and was proposed to be the important site of copy number control. Here we show that the level of repZ expression is rate-limiting for replication and thus copy number, by comparing the levels of repZ expression and copy number from different mutant ColIb-P9 derivatives defective in Inc RNA and pseudoknot formation. Kinetic analyses using in vitro transcribed RNAs indicate that Inc RNA binding and the pseudoknot formation are competitive at the level of initial base paring to loop I. This initial interaction is stimulated by the presence of the loop U residue in the 5' rUUGGCG-3' motif. These results indicate that the competition between the two RNA RNA interactions at the specific site is a novel regulatory mechanism for establishing the constant level of repZ expression and thus copy number. PMID- 9724657 TI - Base excision repair initiation revealed by crystal structures and binding kinetics of human uracil-DNA glycosylase with DNA. AB - Three high-resolution crystal structures of DNA complexes with wild-type and mutant human uracil-DNA glycosylase (UDG), coupled kinetic characterizations and comparisons with the refined unbound UDG structure help resolve fundamental issues in the initiation of DNA base excision repair (BER): damage detection, nucleotide flipping versus extrahelical nucleotide capture, avoidance of apurinic/apyrimidinic (AP) site toxicity and coupling of damage-specific and damage-general BER steps. Structural and kinetic results suggest that UDG binds, kinks and compresses the DNA backbone with a 'Ser-Pro pinch' and scans the minor groove for damage. Concerted shifts in UDG simultaneously form the catalytically competent active site and induce further compression and kinking of the double stranded DNA backbone only at uracil and AP sites, where these nucleotides can flip at the phosphate-sugar junction into a complementary specificity pocket. Unexpectedly, UDG binds to AP sites more tightly and more rapidly than to uracil containing DNA, and thus may protect cells sterically from AP site toxicity. Furthermore, AP-endonuclease, which catalyzes the first damage-general step of BER, enhances UDG activity, most likely by inducing UDG release via shared minor groove contacts and flipped AP site binding. Thus, AP site binding may couple damage-specific and damage-general steps of BER without requiring direct protein protein interactions. PMID- 9724660 TI - Palivizumab, a Humanized Respiratory Syncytial Virus Monoclonal Antibody, Reduces Hospitalization From Respiratory Syncytial Virus Infection in High-risk Infants. AB - Objective. To determine the safety and efficacy of prophylaxis with palivizumab in reducing the incidence of hospitalization because of respiratory syncytial virus (RSV) infection in high-risk infants. Methods. A randomized, double-blind, placebo-controlled trial was conducted at 139 centers in the United States, the United Kingdom, and Canada. During the 1996 to 1997 RSV season, 1502 children with prematurity (75th percentile). MDI, PDI, and REEL scores were compared for the three groups using analysis of variance. To evaluate the relative contributions of physiologic stability, intracranial abnormalities, GA, and early postnatal nutritional intakes, multiple regression analyses were performed using cumulative SNAP score, an intraventricular hemorrhage (IVH) score (incorporating IVH and periventricular leukomalacia), GA, and a weight-change score for the first month as independent variables, and MDI, PDI, and REEL quotients as dependent variables. Regression analyses were repeated, with cumulative SNAP subscores for oxygenation, hypotension, acidosis, and hypoxia/ischemia included with IVH score, GA, and first month weight z score change as independent variables, and MDI, PDI, and REEL quotients as dependent variables. RESULTS: The infants with the highest degree of physiologic instability (cumulative SNAP scores greater than the 75th percentile) had significantly lower MDI scores at 1 year of age and lower PDI scores at 1 year and at 2 to 3 years of age than did infants who were more physiologically stable. Sixty-seven percent of infants with cumulative SNAP scores greater than the 75th percentile had neurodevelopmental abnormalities at 2 to 3 years of age (cerebral palsy or delayed mental, motor, or language development). Using multiple regression analyses, higher cumulative SNAP scores, IVH scores, and GA were associated with lower 1-year MDI scores. Higher cumulative SNAP scores and IVH scores were associated with lower 1-year PDI scores. By 2 years, only higher cumulative SNAP scores were significantly associated with lower MDI and PDI scores. With respect to language development, only lower weight-change scores over the first month were significantly associated with poorer receptive language development. Lower weight-change scores over the first month and higher hypotension scores were significantly associated with poorer expressive language development. In the secondary regression analyses, higher IVH score, higher cumulative oxygenation scores, and higher hypoxia/ischemia scores all were significantly associated with lower 1-year MDI scores. By 2 to 3 years of age, only higher oxygenation scores were significantly associated with lower MDI scores. CONCLUSIONS: Prolonged physiologic instability was associated with deleterious neurodevelopmental consequences for extremely premature infants through 2 to 3 years of age, independent of effects of intracranial abnormalities and GA. PMID- 9724684 TI - Hormonal findings in African-American and Caribbean Hispanic girls with premature adrenarche: implications for polycystic ovarian syndrome. AB - BACKGROUND: Premature adrenarche refers to the early maturation of the adrenal zona reticularis such that the resultant modest hyperandrogenism causes the early appearance of pubic hair before the age of 8 years in girls and 9 years in boys. The precise etiology of premature adrenarche is not known. However, recent studies indicate that certain girls with premature adrenarche are at risk of developing functional ovarian hyperandrogenism, polycystic ovarian syndrome, and hyperinsulinism. Caribbean Hispanic women in general are at increased risk of developing polycystic ovarian syndrome, and African-Americans are at increased risk of developing the complications of hyperinsulinism. Previously, girls with premature adrenarche were reported to have androgens in the range found in normal children in the early stages of puberty. We noted that many of our African American and Caribbean Hispanic patients with premature adrenarche had androgens that were much higher than what has been reported previously. OBJECTIVE: This retrospective study was performed to characterize the adrenocorticotropin stimulated androgen response in an African-American and Caribbean Hispanic population of girls with premature adrenarche. METHODOLOGY: The androgen response to adrenocorticotropin stimulation in 72 African-American and Caribbean Hispanic girls with premature adrenarche was compared with those reported for normal girls in early puberty (Tanner stages II and III). The mean age was 6.8 +/- 0.8 years, bone age was 8 +/- 1.5 years, pubic hair was Tanner stages II and III, and body mass index was 18.6 +/- 4. RESULTS: Of the girls, 28% were found to have elevated stimulated 17OHPregnenolone (17OHPreg) levels that were >2 SD units above the mean for normal early pubertal children. The stimulated ratio of 17OHPreg/17OHProgesterone also was elevated in 18% of the girls and showed a modest correlation with body mass index. CONCLUSION: In contrast to previous studies of girls of mixed ethnic backgrounds with premature adrenarche, 28% of the 72 African-American and Caribbean Hispanic girls with premature adrenarche had adrenocorticotropin-stimulated 17OHPreg levels that were significantly higher than those seen in early pubertal girls. Because 17OHPreg hyperresponsiveness has been described previously in women with hirsutism or polycystic ovarian syndrome, the similar finding in many African-American and Caribbean Hispanic girls with premature adrenarche suggests that the two conditions may share a common mechanism for their hyperandrogenism. Therefore, the hyperandrogenism in certain African-American and Caribbean Hispanic girls with premature adrenarche may not be benign and may be the first presentation of polycystic ovarian syndrome. PMID- 9724685 TI - Bilirubin conjugation, reflected by conjugated bilirubin fractions, in glucose-6 phosphate dehydrogenase-deficient neonates: a determining factor in the pathogenesis of hyperbilirubinemia. AB - BACKGROUND AND OBJECTIVE: Glucose-6-phosphate dehydrogenase (G-6-PD) deficiency is frequently associated with neonatal hyperbilirubinemia, which in severe cases may cause kernicterus and death. Because G-6-PD-deficient individuals frequently undergo acute, trigger-induced hemolytic episodes, increased hemolysis has frequently been implied in the pathogenesis of this neonatal hyperbilirubinemia. However, in Sephardic Jewish G-6-PD-deficient neonates, the rate of hemolysis, reflected by blood carboxyhemoglobin values corrected for inspired carbon monoxide, has been shown to be elevated, not only in those who developed hyperbilirubinemia, but also, to a similar extent, in those who remained only moderately jaundiced. Because at any point, serum total bilirubin values reflect a balance between bilirubin production on the one hand and bilirubin conjugation and elimination on the other, we suspected bilirubin conjugation to be a key factor in the pathogenesis of the hyperbilirubinemia. Physiologically, a fraction of conjugated bilirubin refluxes from the hepatocyte to the serum, and accurate determination of serum conjugated bilirubin fractions can be used to mirror intrahepatocytic bilirubin. Using this principle, we previously demonstrated a decreased diconjugated bilirubin fraction in hyperbilirubinemic G-6-PD-deficient neonates compared with hyperbilirubinemic G-6-PD-normal controls, suggesting diminished bilirubin conjugation. This conjugated bilirubin pattern probably reflects the recently described interaction between G-6-PD deficiency and the variant promoter for the gene encoding the bilirubin conjugating enzyme UDP glucuronosyltransferase, as seen in Gilbert's syndrome. Therefore, we postulated that efficiency of bilirubin conjugation is a crucial factor in the development of hyperbilirubinemia in G-6-PD-deficient neonates. We hypothesized that those G 6-PD-deficient neonates who develop hyperbilirubinemia would have decreased bilirubin conjugation ability, whereas those with a more efficient conjugating system would have a lesser degree of bilirubinemia. METHODS: Term, healthy, male, G-6-PD-deficient neonates with no other obvious predisposing cause for hyperbilirubinemia were selected at random when their serum diazo total bilirubin values ranged from 171 to 254 micromol/L (10-14.9 mg/dL). At this point, simultaneous with the diazo bilirubin determination, serum was collected and frozen for high-performance liquid chromatography (HPLC) measurement of serum bilirubin fractions. The infants were followed clinically and with serum diazo bilirubin determinations until they either did not exceed a serum diazo bilirubin value of 254 micromol/L (14.9 mg/dL) (nonhyperbilirubinemic) or until bilirubin values rose above this level (hyperbilirubinemic), by a process of self selection. A method of alkaline methanolysis, followed by reverse-phase HPLC, was used to measure unconjugated bilirubin and the mono- and diconjugated fractions of serum conjugated bilirubin. Total HPLC bilirubin and total conjugated bilirubin values were calculated from these measured bilirubin fractions. Patients also were classified according to the serum total conjugated bilirubin value as low bilirubin conjugators (serum total conjugated bilirubin less than median) or as high bilirubin conjugators (serum total conjugated bilirubin greater than median). The data were analyzed by comparing serum conjugated bilirubin fractions between the hyperbilirubinemic and nonhyperbilirubinemic groups and the risk of developing hyperbilirubinemia in the low bilirubin conjugators, relative to that of the high bilirubin conjugators. RESULTS: Neonates were sampled at 53 +/- 12 and 58 +/- 12 hours for the subsequently hyperbilirubinemic and nonhyperbilirubinemic groups, respectively (NS). Initial (ie, at the time of sampling) serum total diazo bilirubin values (mean +/- SD) were almost identical for the subsequently hyperbilirubinemic and nonhyperbilirubinemic groups (214 +/ PMID- 9724686 TI - Human milk feedings and infection among very low birth weight infants. AB - BACKGROUND: Preterm infants are immunologically immature at birth. Previous studies have demonstrated that human milk protects against infection in full-term infants, but there are few studies of its effect for preterm infants. OBJECTIVE: To examine the effect of human milk feedings on infection incidence among very low birth weight (VLBW) infants during their initial hospitalization. STUDY DESIGN: The sample consisted of 212 consecutive VLBW infants admitted to the Georgetown University Medical Center neonatal intensive care unit (NICU) during 1992-1993 and surviving to receive enteral feeding. Type of feeding (human milk vs formula), presence of infection and sepsis/meningitis (clinical signs and positive cultures for pathogenic organisms), and potential confounding variables were abstracted from medical records. Multiple logistic regression was used to control for confounders. RESULTS: The incidence of infection (human milk [29.3%] vs formula [47.2%]) and sepsis/meningitis (human milk [19.5%] vs formula [32.6%]) differed significantly by type of feeding. Major risk factors for infection were similar in both groups. Human milk feeding was independently correlated with a reduced odds of infection (odds ratio [OR] = 0.43; 95% confidence interval [CI]: 0.23-0.81), controlling for gestational age, 5-minute Apgar score, mechanical ventilation days, and days without enteral feedings; and was independently correlated with a reduced odds of sepsis/meningitis (OR = 0.47, 95% CI:0.23-0. 95), controlling for gestational age, mechanical ventilation days, and days without enteral feedings. CONCLUSIONS: The incidence of any infection and sepsis/meningitis are significantly reduced in human milk-fed VLBW infants compared with exclusively formula-fed VLBW infants. PMID- 9724687 TI - Factors associated with restraint use of children in fatal crashes. AB - OBJECTIVE: This study was designed to characterize the restraint use of children in fatal crashes and to determine factors that were related to child restraint use. METHODS: Crashes in which a vehicle occupant died were selected from the 1994 Fatal Analysis Reporting System data. Restraint use of children (0-9 years) in these crashes was characterized and examined in relation to vehicle and driver characteristics. RESULTS: Restraint use declined with increasing age of children and increasing number of occupants. Restraint use was also lower in older vehicles, in pickups and large vans, between 3 AM and 6 AM, and in rural areas. Driver characteristics associated with lower restraint use of child passengers included unrestrained drivers, younger drivers, and alcohol use at the time of crash. Driver restraint use was the strongest predictor of child restraint use. Differences in restraint use by age of the child, number of occupants, time, urban/rural area, and driver restraint use persisted in a logistic model. CONCLUSION: Restraint use in crashes is substantially lower than that in observational studies, suggesting that these studies are not reflective of high risk conditions for crashes in which children are involved. Child occupant protection counseling must stress restraint use under all conditions of travel. Local, state, and national efforts must be aimed at increasing family occupant protection for the entire family. PMID- 9724688 TI - Leaf initiation. New developments in an expanding field PMID- 9724689 TI - Resistance of nicotiana benthamiana to phytophthora infestans is mediated by the recognition of the elicitor protein INF1 AB - Phytophthora infestans, the agent of potato and tomato late blight disease, produces a 10-kD extracellular protein, INF1 elicitin. INF1 induces a hypersensitive response in a restricted number of plants, particularly those of the genus Nicotiana. In virulence assays with different P. infestans isolates, five Nicotiana species displayed resistance responses. In all of the interactions, after inoculation with P. infestans zoospores, penetration of an epidermal cell was observed, followed by localized necrosis typical of a hypersensitive response. To determine whether INF1 functions as an avirulence factor in these interactions, we adopted a gene-silencing strategy to inhibit INF1 production. Several transformants deficient in inf1 mRNA and INF1 protein were obtained. These strains remained pathogenic on host plants. However, in contrast to the wild-type and control transformant strains, INF1-deficient strains induced disease lesions when inoculated on N. benthamiana. These results demonstrate that the elicitin INF1 functions as an avirulence factor in the interaction between N. benthamiana and P. infestans. PMID- 9724690 TI - Localized upregulation of a new expansin gene predicts the site of leaf formation in the tomato meristem. AB - Expansins are extracellular proteins that increase plant cell wall extensibility in vitro and are thought to be involved in cell expansion. We showed in a previous study that administration of an exogenous expansin protein can trigger the initiation of leaflike structures on the shoot apical meristem of tomato. Here, we studied the expression patterns of two tomato expansin genes, LeExp2 and LeExp18. LeExp2 is preferentially expressed in expanding tissues, whereas LeExp18 is expressed preferentially in tissues with meristematic activity. In situ hybridization experiments showed that LeExp18 expression is elevated in a group of cells, called I1, which is the site of incipient leaf primordium initiation. Thus, LeExp18 expression is a molecular marker for leaf initiation, predicting the site of primordium formation at a time before histological changes can be detected. We propose a model for the regulation of phyllotaxis that postulates a crucial role for expansin in leaf primordium initiation. PMID- 9724691 TI - A mutation within the leucine-rich repeat domain of the Arabidopsis disease resistance gene RPS5 partially suppresses multiple bacterial and downy mildew resistance genes. AB - Recognition of pathogens by plants is mediated by several distinct families of functionally variable but structurally related disease resistance (R) genes. The largest family is defined by the presence of a putative nucleotide binding domain and 12 to 21 leucine-rich repeats (LRRs). The function of these LRRs has not been defined, but they are speculated to bind pathogen-derived ligands. We have isolated a mutation in the Arabidopsis RPS5 gene that indicates that the LRR region may interact with other plant proteins. The rps5-1 mutation causes a glutamate-to-lysine substitution in the third LRR and partially compromises the function of several R genes that confer bacterial and downy mildew resistance. The third LRR is relatively well conserved, and we speculate that it may interact with a signal transduction component shared by multiple R gene pathways. PMID- 9724692 TI - Temperature-sensitive splicing in the floral homeotic mutant apetala3-1. AB - The floral homeotic gene APETALA3 (AP3) is required for stamen and petal development in Arabidopsis. The previously described ap3-1 allele is temperature sensitive and carries a missense mutation near a 5' splice site. The missense mutation lies within a domain of the AP3 protein that is thought to be important for protein-protein interactions, which suggests that temperature sensitivity of ap3-1 could reflect an unstable interaction with cofactors. Here, we show instead that the ap3-1 mutation causes a temperature-dependent splicing defect and that temperature sensitivity is not a property of the protein products of ap3-1 but of RNA processing, possibly because of unstable base pairing between the transcript and small nuclear RNAs. The unexpected defect of the ap3-1 mutant offers unique opportunities for genetic and molecular studies of splice site recognition in plants. PMID- 9724693 TI - An intragenic suppressor of the Arabidopsis floral organ identity mutant apetala3 1 functions by suppressing defects in splicing. AB - The Arabidopsis floral organ identity gene APETALA3 (AP3) specifies the identity of petals and stamens in the flower. In flowers mutant for the temperature sensitive ap3-1 allele, the petals and stamens are partially converted to sepals and carpels, respectively. ap3-1 contains a single nucleotide change in the AP3 gene that alters both an amino acid in the AP3 protein and the 5' splice consensus site for intron 5. Surprisingly, the Ap3-1 mutant phenotype is not due to the missense mutation but instead is due to defects in splicing; specifically, exon 5 is frequently skipped by the splicing machinery at the restrictive temperature. In a screen for suppressors of ap3-1, we isolated an intragenic suppressor, ap3-11, that functions to suppress the splicing defects of ap3-1. Using a reverse transcriptase-polymerase chain reaction assay, we demonstrate that the percentage of full-length exon 5-containing AP3 RNAs correlates with the phenotype of the flowers in both ap3-1 and ap3-11. Rather surprisingly, the ap3 11 suppressor mutation is located in intron 4. One model explaining the function of ap3-11 is that the ap3-11 suppressor creates a novel branch point sequence that causes exon 5 to be more frequently recognized by the splicing machinery. The identification of such a suppressor strongly suggests that exon-scanning models of intron-exon recognition are operative in plants. PMID- 9724694 TI - Phytochrome E influences internode elongation and flowering time in Arabidopsis. AB - From a screen of M2 seedlings derived from gamma-mutagenesis of seeds doubly null for phytochromes phyA and phyB, we isolated a mutant lacking phyE. The PHYE gene of the selected mutant, phyE-1, was found to contain a 1-bp deletion at a position equivalent to codon 726, which is predicted to result in a premature stop at codon 739. Immunoblot analysis showed that the phyE protein was undetectable in the phyE-1 mutant. In the phyA- and phyB-deficient background, phyE deficiency led to early flowering, elongation of internodes between adjacent rosette leaves, and reduced petiole elongation. This is a phenocopy of the response of phyA phyB seedlings to end-of-day far-red light treatments. Furthermore, a phyE deficiency attenuated the responses of phyA phyB seedlings to end-of-day far-red light treatments. Monogenic phyE mutants were indistinguishable from wild-type seedlings. However, phyB phyE double mutants flowered earlier and had longer petioles than did phyB mutants. The elongation and flowering responses conferred by phyE deficiency are typical of shade avoidance responses to the low red/far-red ratio. We conclude that in conjunction with phyB and to a lesser extent with phyD, phyE functions in the regulation of the shade avoidance syndrome. PMID- 9724695 TI - Cyanide restores N gene-mediated resistance to tobacco mosaic virus in transgenic tobacco expressing salicylic acid hydroxylase AB - Salicylhydroxamic acid (SHAM), an inhibitor of alternative oxidase (AOX), blocks salicylic acid-induced resistance to tobacco mosaic virus (TMV) but does not inhibit pathogenesis-related PR-1 protein synthesis or resistance to fungal and bacterial pathogens. We found that the synthetic resistance-inducing chemical 2, 6-dichloroisonicotinic acid also induced Aox transcript accumulation and SHAM sensitive resistance to TMV. The respiratory inhibitors antimycin A and KCN also induced Aox transcript accumulation and resistance to TMV but did not induce PR-1 accumulation. Tobacco plants of the TMV-resistant cultivar Samsun NN transformed with the salicylic acid hydroxylase (nahG) gene could no longer restrict virus to the inoculation site, resulting in spreading necrosis instead of discrete necrotic lesions. Treatment with KCN restored TMV localization and normal lesion morphology. SHAM antagonized this effect, allowing virus escape and spreading necrosis to resume. The results demonstrate the importance of the SHAM-sensitive (potentially AOX-dependent) signal transduction pathway in mediating virus localization early in the hypersensitive response. PMID- 9724696 TI - Relocation of a Ca2+-dependent protein kinase activity during pollen tube reorientation AB - Pollen tube reorientation is a dynamic cellular event that is crucial for successful fertilization. We have shown previously that pollen tube orientation is regulated by cytosolic free calcium ([Ca2+]c). In this paper, we studied the activity of a Ca2+-dependent protein kinase during reorientation. The kinase activity was assayed in living cells by using confocal ratio imaging of BODIPY FL bisindolylmaleimide. We found that growing pollen tubes exhibited higher protein kinase activity in the apical region, whereas nongrowing cells showed uniform distribution. Modification of growth direction by diffusion of inhibitors/activators from a micropipette showed the spatial redistribution of kinase activity to predict the new growth orientation. Localized increases in [Ca2+]c induced by photolysis of caged Ca2+ that led to reorientation also increased kinase activity. Molecular and immunological assays suggest that this kinase may show some functional homology with protein kinase C. We suggest that the tip-localized gradient of kinase activity promotes Ca2+-mediated exocytosis and may act to regulate Ca2+ channel activity. PMID- 9724697 TI - A recessive heterochronic mutation, plastochron1, shortens the plastochron and elongates the vegetative phase in rice AB - We describe two recessive alleles of a rice heterochronic gene, plastochron1-1 (pla1-1) and pla1-2, that reduce the length of the plastochron to approximately half that of the wild type. Because the onset of the reproductive phase in pla1 was not temporally affected, the number of leaves produced in the vegetative phase was nearly twice that produced in the wild type. Panicle development was severely disturbed in pla1 mutants. In pla1-1, many primordia of primary rachis branches were converted into vegetative shoots. These ectopic shoots repeated the initiation of panicle development and the conversion of primary rachis branches into shoots. In the weak allele pla1-2, however, only the basal one or two primordia developed as vegetative shoots, and the remaining primordia developed to produce a truncated panicle. These results indicate that both vegetative and reproductive programs are expressed simultaneously during the reproductive phase of pla1; however, the degree varied depending on the strength of the allele. Accordingly, pla1 is a heterochronic mutation that extends the vegetative period. The shoot apical meristem of pla1 was larger than that of the wild type, although the shape was not modified. An in situ hybridization experiment using the histone H4 gene as a probe revealed that cell divisions are accelerated in the pla1 meristem. The PLA1 gene is considered to regulate the duration of the vegetative phase by controlling the rate of leaf production in the meristem. PMID- 9724699 TI - Glutathione metabolic genes coordinately respond to heavy metals and jasmonic acid in Arabidopsis. AB - Glutathione plays a pivotal role in protecting plants from environmental stresses, oxidative stress, xenobiotics, and some heavy metals. Arabidopsis plants treated with cadmium or copper responded by increasing transcription of the genes for glutathione synthesis, gamma-glutamylcysteine synthetase and glutathione synthetase, as well as glutathione reductase. The response was specific for those metals whose toxicity is thought to be mitigated through phytochelatins, and other toxic and nontoxic metals did not alter mRNA levels. Feeding experiments suggested that neither oxidative stress, as results from exposure to H2O2, nor oxidized or reduced glutathione levels were responsible for activating transcription of these genes. Jasmonic acid also activated the same suite of genes, which suggests that it might be involved in the signal transduction pathway for copper and cadmium. Jasmonic acid treatment increased mRNA levels and the capacity for glutathione synthesis but did not alter the glutathione content in unstressed plants, which supports the idea that the glutathione concentration is controlled at multiple levels. PMID- 9724698 TI - PIOX, a new pathogen-induced oxygenase with homology to animal cyclooxygenase. AB - Changes in gene expression induced in tobacco leaves by the harpin HrpN protein elicitor were examined, and a new cDNA, piox (for pathogen-induced oxygenase), with homology to genes encoding cyclooxygenase or prostaglandin endoperoxide synthase (PGHS), was identified. In addition to the amino acid identity determined, the protein encoded by piox is predicted to have a structural core similar to that of ovine PGHS-1. Moreover, studies of protein functionality demonstrate that the PIOX recombinant protein possesses at least one of the two enzymatic activities of PGHSs, that of catalyzing the oxygenation of polyunsaturated fatty acids. piox transcripts accumulated after protein elicitor treatment or inoculation with bacteria. Expression of piox was induced in tissues responding to inoculation with both incompatible and compatible bacteria, but RNA and protein accumulation differed for both types of interactions. We show that expression of piox is rapidly induced in response to various cellular signals mediating plant responses to pathogen infection and that activation of piox expression is most likely related to the oxidative burst that takes place during the cell death processes examined. Cyclooxygenase catalyzes the first committed step in the formation of prostaglandins and thromboxanes, which are lipid-derived signal molecules that mediate many cellular processes, including the immune response in vertebrates. The finding of tobacco PIOX suggests that more similarities than hitherto expected will be found between the lipid-based responses for plant and animal systems. PMID- 9724700 TI - Molecular localization of a redox-modulated process regulating plant mitochondrial electron transport AB - Using in organellar assays, we found that significant tobacco alternative oxidase (AOX) activity is dependent on both reduction of a putative regulatory disulfide bond and the presence of pyruvate, which may interact with a Cys sulfhydryl. This redox modulation and pyruvate activation thus may be important in determining the partitioning of electrons to AOX in vivo. To investigate these regulatory mechanisms, we generated tobacco plants expressing mutated AOX proteins. Mutation of the most N-terminal Cys residue (Cys-126) to an Ala residue produced an AOX that could not be converted to the disulfide-linked form, thus identifying this Cys residue as being responsible for redox modulation. Although this mutation might be expected to produce an AOX with constitutive high activity in the presence of pyruvate, we found it to have minimal in organellar activity in the presence of pyruvate. Nonetheless, the Cys-126 mutation did not appear to have compromised the catalytic function of AOX, given that cells expressing the protein displayed high rates of cyanide-resistant respiration in vivo. The striking difference between in vivo and in organellar results suggests that an additional mechanism(s), as yet unidentified by in organellar assays, may promote activity in vivo. Mutation of the Cys residue nearest the presumptive active site (Cys-176) to an Ala residue did not prevent disulfide bond formation or affect the ability of AOX to be stimulated by pyruvate, indicating that this Cys residue is involved in neither redox modulation nor pyruvate activation. PMID- 9724701 TI - The plant wound hormone systemin binds with the N-terminal part to its receptor but needs the C-terminal part to activate it. AB - Suspension-cultured cells of Lycopersicon peruvianum respond with rapid medium alkalinization and a strong increase of a MAP kinase-like activity when treated with subnanomolar concentrations of the plant wound hormone systemin. Systemin fragments comprising the N-terminal 14 amino acids (syst1-14) or the C-terminal four amino acids (syst15-18), added singly or in combination, were inactive as inducers of these responses. Syst1-14 but not syst15-18 antagonized activity of intact systemin in a competitive manner. Likewise, intact systemin showed stimulatory, syst1-14 antagonistic activity, and syst15-18 showed no activity in leaf pieces of tomato (L. esculentum) plants assayed for the induction of ethylene biosynthesis. To study the molecular basis of perception, we extended the C-terminal end of systemin by a tyrosine residue and radioiodinated it to yield systemin-125I-iodotyrosine. In membrane preparations of L. peruvianum, this radioligand exhibited rapid, saturable, and reversible binding to a single class of binding sites. Binding showed a dissociation constant of approximately 1 nM, and binding of radioligand was efficiently competed by unlabeled systemin but not by syst15-18 or structurally unrelated peptides. Binding was also competed by the systemin antagonists syst1-14 and syst-Ala-17 (IC50 of 500 and 1000 nM, respectively). Thus, this binding site exhibits the characteristics expected for a functional systemin receptor. Based on these results, we propose a two-step mechanism for systemin action, with binding of the N-terminal part to the receptor as the first step and activation of responses with the C-terminal part as the second step. PMID- 9724702 TI - A novel signaling pathway controlling induced systemic resistance in Arabidopsis. AB - Plants have the ability to acquire an enhanced level of resistance to pathogen attack after being exposed to specific biotic stimuli. In Arabidopsis, nonpathogenic, root-colonizing Pseudomonas fluorescens bacteria trigger an induced systemic resistance (ISR) response against infection by the bacterial leaf pathogen P. syringae pv tomato. In contrast to classic, pathogen-induced systemic acquired resistance (SAR), this rhizobacteria-mediated ISR response is independent of salicylic acid accumulation and pathogenesis-related gene activation. Using the jasmonate response mutant jar1, the ethylene response mutant etr1, and the SAR regulatory mutant npr1, we demonstrate that signal transduction leading to P. fluorescens WCS417r-mediated ISR requires responsiveness to jasmonate and ethylene and is dependent on NPR1. Similar to P. fluorescens WCS417r, methyl jasmonate and the ethylene precursor 1 aminocyclopropane-1-carboxylate were effective in inducing resistance against P. s. tomato in salicylic acid-nonaccumulating NahG plants. Moreover, methyl jasmonate-induced protection was blocked in jar1, etr1, and npr1 plants, whereas 1-aminocyclopropane-1-carboxylate-induced protection was affected in etr1 and npr1 plants but not in jar1 plants. Hence, we postulate that rhizobacteria mediated ISR follows a novel signaling pathway in which components from the jasmonate and ethylene response are engaged successively to trigger a defense reaction that, like SAR, is regulated by NPR1. We provide evidence that the processes downstream of NPR1 in the ISR pathway are divergent from those in the SAR pathway, indicating that NPR1 differentially regulates defense responses, depending on the signals that are elicited during induction of resistance. PMID- 9724703 TI - Nitric oxide in plant immunity. PMID- 9724704 TI - Skiing the black diamond slope: progress on the biochemistry of translesion DNA synthesis. PMID- 9724705 TI - Diverse RNA substrates for aminoacylation: clues to origins? PMID- 9724706 TI - The central questions in pain perception may be peripheral. PMID- 9724707 TI - Altering the biochemical state of individual cultured cells and organelles with ultramicroelectrodes. AB - We describe an efficient technique for the selective chemical and biological manipulation of the contents of individual cells. This technique is based on the electric-field-induced permeabilization (electroporation) in biological membranes using a low-voltage pulse generator and microelectrodes. A spatially highly focused electric field allows introduction of polar cell-impermeant solutes such as fluorescent dyes, fluorogenic reagents, and DNA into single cells. The high spatial resolution of the technique allows for design of, for example, cellular network constructions in which cells in close contact with each other can be made to possess different biochemical, biophysical, and morphological properties. Fluorescein, and fluo-3 (a calcium-sensitive fluorophore), are electroporated into the soma of cultured single progenitor cells derived from adult rat hippocampus. Fluo-3 also is introduced into individual submicrometer diameter processes of thapsigargin-treated progenitor cells, and a plasmid vector cDNA construct (pRAY 1), expressing the green fluorescent protein, is electroporated into cultured single COS 7 cells. At high electric field strengths, observations of dye-transfer into organelles are proposed. PMID- 9724708 TI - Photochemical protease: site-specific photocleavage of hen egg lysozyme and bovine serum albumin. AB - Site-specific photocleavage of hen egg lysozyme and bovine serum albumin (BSA) by N-(l-phenylalanine)-4-(1-pyrene)butyramide (Py-Phe) is reported. Py-Phe binds to lysozyme and BSA with binding constants 2.2 +/- 0.3 x 10(5) M-1 and 6.5 +/- 0.4 x 10(7) M-1, respectively. Photocleavage of lysozyme and BSA was achieved with high specificity when a mixture of protein, Py-Phe, and an electron acceptor, cobalt(III) hexammine (CoHA), was irradiated at 344 nm. Quantum yields of photocleavage of lysozyme and BSA were 0.26 and 0.0021, respectively. No protein cleavage was observed in the absence of Py-Phe, CoHA, or light. N-terminal sequencing of the protein fragments indicated a single cleavage site of lysozyme between Trp-108 and Val-109, whereas the cleavage of BSA was found to be between Leu-346 and Arg-347. Laser flash photolysis studies of a mixture of protein, Py Phe, and CoHA showed a strong transient with absorption centered at approximately 460 nm, corresponding to pyrene cation radical. Quenching of the singlet excited state of Py-Phe by CoHA followed by the reaction of the resulting pyrenyl cation radical with the protein backbone may be responsible for the protein cleavage. The high specificity of photocleavage may be valuable in targeting specific sites of proteins with small molecules. PMID- 9724709 TI - A recipe for randomness. AB - Despite many diverse theories that address closely related themes-e. g., probability theory, algorithmic complexity, cryptoanalysis, and pseudorandom number generation-a near-void remains in constructive methods certified to yield the desired "random" output. Herein, we provide explicit techniques to produce broad sets of both highly irregular finite and normal infinite sequences, based on constructions and properties derived from approximate entropy (ApEn), a computable formulation of sequential irregularity. Furthermore, for infinite sequences, we considerably refine normality, by providing methods for constructing diverse classes of normal numbers, classified by the extent to which initial segments deviate from maximal irregularity. PMID- 9724710 TI - Rapid polyether cleavage via extracellular one-electron oxidation by a brown-rot basidiomycete. AB - Fungi that cause brown rot of wood are essential biomass recyclers and also the principal agents of decay in wooden structures, but the extracellular mechanisms by which they degrade lignocellulose remain unknown. To test the hypothesis that brown-rot fungi use extracellular free radical oxidants as biodegradative tools, Gloeophyllum trabeum was examined for its ability to depolymerize an environmentally recalcitrant polyether, poly(ethylene oxide) (PEO), that cannot penetrate cell membranes. Analyses of degraded PEOs by gel permeation chromatography showed that the fungus cleaved PEO rapidly by an endo route. 13C NMR analyses of unlabeled and perdeuterated PEOs recovered from G. trabeum cultures showed that a major route for depolymerization was oxidative C---C bond cleavage, a reaction diagnostic for hydrogen abstraction from a PEO methylene group by a radical oxidant. Fenton reagent (Fe(II)/H2O2) oxidized PEO by the same route in vitro and therefore might account for PEO biodegradation if it is produced by the fungus, but the data do not rule out involvement of less reactive radicals. The reactivity and extrahyphal location of this PEO-degrading system suggest that its natural function is to participate in the brown rot of wood and that it may enable brown-rot fungi to degrade recalcitrant organopollutants. PMID- 9724711 TI - Nitric oxide dioxygenase: an enzymic function for flavohemoglobin. AB - Nitric oxide (NO*) is a toxin, and various life forms appear to have evolved strategies for its detoxification. NO*-resistant mutants of Escherichia coli were isolated that rapidly consumed NO*. An NO*-converting activity was reconstituted in extracts that required NADPH, FAD, and O2, was cyanide-sensitive, and produced NO3-. This nitric oxide dioxygenase (NOD) contained 19 of 20 N-terminal amino acids identical to those of the E. coli flavohemoglobin. Furthermore, NOD activity was produced by the flavohemoglobin gene and was inducible by NO*. Flavohemoglobin/NOD-deficient mutants were also sensitive to growth inhibition by gaseous NO*. The results identify a function for the evolutionarily conserved flavohemoglobins and, moreover, suggest that NO* detoxification may be a more ancient function for the widely distributed hemoglobins, and associated methemoglobin reductases, than dioxygen transport and storage. PMID- 9724712 TI - Quantitative assessment of enzyme specificity in vivo: P2 recognition by Kex2 protease defined in a genetic system. AB - The specificity of the yeast proprotein-processing Kex2 protease was examined in vivo by using a sensitive, quantitative assay. A truncated prepro-alpha-factor gene encoding an alpha-factor precursor with a single alpha-factor repeat was constructed with restriction sites for cassette mutagenesis flanking the single Kex2 cleavage site (-SLDKR downward arrowEAEA-). All of the 19 substitutions for the Lys (P2) residue in the cleavage site were made. The wild-type and mutant precursors were expressed in a yeast strain lacking the chromosomal genes encoding Kex2 and prepro-alpha-factor. Cleavage of the 20 sites by Kex2, expressed at the wild-type level, was assessed by using a quantitative-mating assay with an effective range greater than six orders of magnitude. All substitutions for Lys at P2 decreased mating, from 2-fold for Arg to >10(6)-fold for Trp. Eviction of the Kex2-encoding plasmid indicated that cleavage of mutant sites by other cellular proteases was not a complicating factor. Mating efficiencies of strains expressing the mutant precursors correlated well with the specificity (kcat/KM) of purified Kex2 for comparable model peptide substrates, validating the in vivo approach as a quantitative method. The results support the conclusion that KM, which is heavily influenced by the nature of the P2 residue, is a major determinant of cleavage efficiency in vivo. P2 preference followed the rank order: Lys > Arg > Thr > Pro > Glu > Ile > Ser > Ala > Asn > Val > Cys > AsP > Gln > Gly > His > Met > Leu > Tyr > Phe > Trp. PMID- 9724713 TI - Overexpression of the large subunit of the protein Ku suppresses metallothionein I induction by heavy metals. AB - Metallothioneins (MT) are involved in the scavenging of the toxic heavy metals and protection of cells from reactive oxygen intermediates. To investigate the potential role of the protein Ku in the expression of MT, we measured the level of MT-I mRNA in the parental rat fibroblast cell line (Rat 1) and the cell lines that stably and constitutively overexpress the small subunit, the large subunit, and the heterodimer of Ku. Treatment with CdS04 or ZnS04 elevated the MT-I mRNA level 20- to 30-fold in the parental cells and the cells (Ku-70) that overproduce the small subunit or those (Ku-7080) overexpressing the heterodimer. By contrast, the cells (Ku-80) overexpressing the large subunit of Ku failed to induce MT-I. In vitro transcription assay showed that the MT-I promoter activity was suppressed selectively in the nuclear extracts from Ku-80 cells. The specificity of the repressor function was shown by the induction of hsp 70, another Cd inducible gene, in Ku-80 cells. Addition of the nuclear extract from Ku-80 cells at the start of the transcription reaction abolished the MT-l promoter activity in the Rat 1 cell extract. The transcript once formed in Rat 1 nuclear extract was not degraded by further incubation with the extract from Ku-80 cells. The repressor was sensitive to heat. The DNA-binding activities of at least four transcription factors that control the MT-I promoter activity were not affected in Ku-80 cells. These observations have set the stage for further exploration of the mechanisms by which the Ku subunit mediates suppression of MT induction. PMID- 9724714 TI - Evolution of an enzyme active site: the structure of a new crystal form of muconate lactonizing enzyme compared with mandelate racemase and enolase. AB - Muconate lactonizing enzyme (MLE), a component of the beta-ketoadipate pathway of Pseudomonas putida, is a member of a family of related enzymes (the "enolase superfamily") that catalyze the abstraction of the alpha-proton of a carboxylic acid in the context of different overall reactions. New untwinned crystal forms of MLE were obtained, one of which diffracts to better than 2.0-A resolution. The packing of the octameric enzyme in this crystal form is unusual, because the asymmetric unit contains three subunits. The structure of MLE presented here contains no bound metal ion, but is very similar to a recently determined Mn2+ bound structure. Thus, absence of the metal ion does not perturb the structure of the active site. The structures of enolase, mandelate racemase, and MLE were superimposed. A comparison of metal ligands suggests that enolase may retain some characteristics of the ancestor of this enzyme family. Comparison of other residues involved in catalysis indicates two unusual patterns of conservation: (i) that the position of catalytic atoms remains constant, although the residues that contain them are located at different points in the protein fold; and (ii) that the positions of catalytic residues in the protein scaffold are conserved, whereas their identities and roles in catalysis vary. PMID- 9724715 TI - Defective insulin secretion and enhanced insulin action in KATP channel-deficient mice. AB - ATP-sensitive K+ (KATP) channels regulate many cellular functions by linking cell metabolism to membrane potential. We have generated KATP channel-deficient mice by genetic disruption of Kir6.2, which forms the K+ ion-selective pore of the channel. The homozygous mice (Kir6.2(-/-)) lack KATP channel activity. Although the resting membrane potential and basal intracellular calcium concentrations ([Ca2+]i) of pancreatic beta cells in Kir6.2(-/-) are significantly higher than those in control mice (Kir6.2(+/+)), neither glucose at high concentrations nor the sulfonylurea tolbutamide elicits a rise in [Ca2+]i, and no significant insulin secretion in response to either glucose or tolbutamide is found in Kir6.2(-/-), as assessed by perifusion and batch incubation of pancreatic islets. Despite the defect in glucose-induced insulin secretion, Kir6.2(-/-) show only mild impairment in glucose tolerance. The glucose-lowering effect of insulin, as assessed by an insulin tolerance test, is increased significantly in Kir6.2(-/-), which could protect Kir6.2(-/-) from developing hyperglycemia. Our data indicate that the KATP channel in pancreatic beta cells is a key regulator of both glucose and sulfonylurea-induced insulin secretion and suggest also that the KATP channel in skeletal muscle might be involved in insulin action. PMID- 9724716 TI - Ribonuclease A variants with potent cytotoxic activity. AB - Select members of the bovine pancreatic ribonuclease A (RNase A) superfamily are potent cytotoxins. These cytotoxic ribonucleases enter the cytosol, where they degrade cellular RNA and cause cell death. Ribonuclease inhibitor (RI), a cytosolic protein, binds to members of the RNase A superfamily with inhibition constants that span 10 orders of magnitude. Here, we show that the affinity of a ribonuclease for RI plays an integral role in defining the potency of a cytotoxic ribonuclease. RNase A is not cytotoxic and binds RI with high affinity. Onconase, a cytotoxic RNase A homolog, binds RI with low affinity. To disrupt the RI-RNase A interaction, three RNase A residues (Asp-38, Gly-88, and Ala-109) that form multiple contacts with RI were replaced with arginine. Replacing Asp-38 and Ala 109 with an arginine residue has no effect on the RI-RNase interaction. In addition, these variants are not cytotoxic. In contrast, replacing Gly-88 with an arginine residue yields a ribonuclease (G88R RNase A) that retains catalytic activity in the presence of RI and is cytotoxic to a transformed cell line. Replacing Gly-88 with aspartate also yields a ribonuclease (G88D RNase A) with a decreased affinity for RI and cytotoxic activity. The cytotoxic potency of onconase, G88R RNase A, and G88D RNase A correlate with RI evasion. We conclude that ribonucleases that retain catalytic activity in the presence of RI are cytotoxins. This finding portends the development of a class of chemotherapeutic agents based on pancreatic ribonucleases. PMID- 9724717 TI - A novel DNA-binding motif in MarA: the first structure for an AraC family transcriptional activator. AB - A crystal structure for a member of the AraC prokaryotic transcriptional activator family, MarA, in complex with its cognate DNA-binding site is described. MarA consists of two similar subdomains, each containing a helix-turn helix DNA-binding motif. The two recognition helices of the motifs are inserted into adjacent major groove segments on the same face of the DNA but are separated by only 27 A thereby bending the DNA by approximately 35 degrees. Extensive interactions between the recognition helices and the DNA major groove provide the sequence specificity. PMID- 9724718 TI - Crystal structures of eukaryotic translation initiation factor 5A from Methanococcus jannaschii at 1.8 A resolution. AB - Eukaryotic translation initiation factor 5A (eIF-5A) is a ubiquitous protein found in all eukaryotic cells. The protein is closely associated with cell proliferation in the G1-S stage of the cell cycle. Recent findings show that the eIF-5A proteins are highly expressed in tumor cells and act as a cofactor of the Rev protein in HIV-1-infected cells. The mature eIF is the only protein known to have the unusual amino acid hypusine, a post-translationally modified lysine. The crystal structure of eIF-5A from Methanococcus jannaschii (MJ eIF-5A) has been determined at 1.9 A and 1.8 A resolution in two crystal forms by using the multiple isomorphous replacement method and the multiwavelength anomalous diffraction method for the first crystal form and the molecular replacement method for the second crystal form. The structure consists of two folding domains, one of which is similar to the oligonucleotide-binding domain found in the prokaryotic cold shock protein and the translation initiation factor IF1 despite the absence of any significant sequence similarities. The 12 highly conserved amino acid residues found among eIF-5As include the hypusine site and form a long protruding loop at one end of the elongated molecule. PMID- 9724719 TI - Protein kinase A-catalyzed phosphorylation of heat shock protein 60 chaperone regulates its attachment to histone 2B in the T lymphocyte plasma membrane. AB - Accumulating evidence suggests that the mitochondrial molecular chaperone heat shock protein 60 (hsp60) also can localize in extramitochondrial sites. However, direct evidence that hsp60 functions as a chaperone outside of mitochondria is presently lacking. A 60-kDa protein that is present in the plasma membrane of a human leukemic CD4(+) CEM-SS T cell line and is phosphorylated by protein kinase A (PKA) was identified as hsp60. An 18-kDa plasma membrane-associated protein coimmunoprecipitated with hsp60 and was identified as histone 2B (H2B). Hsp60 physically associated with H2B when both molecules were in their dephospho forms. By contrast, PKA-catalyzed phosphorylation of both hsp60 and H2B caused dissociation of H2B from hsp60 and loss of H2B from the plasma membrane of intact T cells. These results suggest that (i) hsp60 and H2B can localize in the T cell plasma membrane; (ii) hsp60 functions as a molecular chaperone for H2B; and (iii) PKA-catalyzed phosphorylation of both hsp60 and H2B appears to regulate the attachment of H2B to hsp60. We propose a model in which phosphorylation/dephosphorylation regulates chaperoning of H2B by hsp60 in the plasma membrane. PMID- 9724720 TI - Resected RNA pseudoknots and their recognition by histidyl-tRNA synthetase. AB - Duplexes constituted by closed or open RNA circles paired to single-stranded oligonucleotides terminating with 3'-CCAOH form resected pseudoknots that are substrates of yeast histidyl-tRNA synthetase. Design of this RNA fold is linked to the mimicry of the pseudoknotted amino acid accepting branch of the tRNA-like domain from brome mosaic virus, known to be charged by tyrosyl-tRNA synthetases, with RNA minihelices recapitulating accepting branches of canonical tRNAs. Prediction of the histidylation function of the new family of minimalist tRNA like structures relates to the geometry of resected pseudoknots that allows proper presentation to histidyl-tRNA synthetase of analogues of the histidine identity determinants N-1 and N73 present in tRNAs. This geometry is such that the analogue of the major N-1 histidine determinant in the RNA circles faces the analogue of the discriminator N73 nucleotide in the accepting oligonucleotides. The combination of identity elements found in tRNAHis species from archaea, eubacteria, and organelles (G-1/C73) is the most efficient for determining histidylation of the duplexes. The inverse combination (C-1/G73) leads to the worst histidine acceptors with charging efficiencies reduced by 2-3 orders of magnitude. Altogether, these findings open new perspectives for understanding evolution of tRNA identity and serendipitous RNA functions. PMID- 9724721 TI - Redesigning an FKBP-ligand interface to generate chemical dimerizers with novel specificity. AB - FKBP ligand homodimers can be used to activate signaling events inside cells and animals that have been engineered to express fusions between appropriate signaling domains and FKBP. However, use of these dimerizers in vivo is potentially limited by ligand binding to endogenous FKBP. We have designed ligands that bind specifically to a mutated FKBP over the wild-type protein by remodeling an FKBP-ligand interface to introduce a specificity binding pocket. A compound bearing an ethyl substituent in place of a carbonyl group exhibited sub nanomolar affinity and 1,000-fold selectivity for a mutant FKBP with a compensating truncation of a phenylalanine residue. Structural and functional analysis of the new pocket showed that recognition is surprisingly relaxed, with the modified ligand only partially filling the engineered cavity. We incorporated the specificity pocket into a fusion protein containing FKBP and the intracellular domain of the Fas receptor. Cells expressing this modified chimeric protein potently underwent apoptosis in response to AP1903, a homodimer of the modified ligand, both in culture and when implanted into mice. Remodeled dimerizers such as AP1903 are ideal reagents for controlling the activities of cells that have been modified by gene therapy procedures, without interference from endogenous FKBP. PMID- 9724722 TI - Zinc-dependent dimers observed in crystals of human endostatin. AB - The crystal structure of human endostatin reveals a zinc-binding site. Atomic absorption spectroscopy indicates that zinc is a constituent of both human and murine endostatin in solution. The human endostatin zinc site is formed by three histidines at the N terminus, residues 1, 3, and, 11, and an aspartic acid at residue 76. The N-terminal loop ordered around the zinc makes a dimeric contact in human endostatin crystals. The location of the zinc site at the amino terminus, immediately adjacent to the precursor cleavage site, suggests the possibility that the zinc may be involved in activation of the antiangiogenic activity following cleavage from the inactive collagen XVIII precursor or in the cleavage process itself. PMID- 9724723 TI - Altering the anaerobic transcription factor FNR confers a hemolytic phenotype on Escherichia coli K12. AB - The recent outbreaks of Escherichia coli O157-associated food poisoning have focused attention on the virulence determinants of E. coli. Here, it is reported that single base substitutions in the fnr gene encoding the oxygen-responsive transcription regulator FNR (fumarate and nitrate reduction regulator) are sufficient to confer a hemolytic phenotype on E. coli K12, the widely used laboratory strain. The mechanism involves enhancing the expression of a normally dormant hemolysin gene (hlyE) located in the E. coli chromosome. The mutations direct single amino acid substitutions in the activating regions (AR1 and AR3) of FNR that contact RNA polymerase. It is concluded that altering a resident transcription regulator, or acquisition of a competent heterologous regulator, could generate a pool of hemolytic, and therefore more virulent, strains of E. coli in nature. PMID- 9724724 TI - Substrate recruitment to cyclin-dependent kinase 2 by a multipurpose docking site on cyclin A. AB - An important question in the cell cycle field is how cyclin-dependent kinases (cdks) target their substrates. We have studied the role of a conserved hydrophobic patch on the surface of cyclin A in substrate recognition by cyclin A cdk2. This hydrophobic patch is approximately 35A away from the active site of cdk2 and contains the MRAIL sequence conserved among a number of mammalian cyclins. In the x-ray structure of cyclin A-cdk2-p27, this hydrophobic patch contacts the RNLFG sequence in p27 that is common to a number of substrates and inhibitors of mammalian cdks. We find that mutation of this hydrophobic patch on cyclin A eliminates binding to proteins containing RXL motifs without affecting binding to cdk2. This docking site is critical for cyclin A-cdk2 phosphorylation of substrates containing RXL motifs, but not for phosphorylation of histone H1. Impaired substrate binding by the cyclin is the cause of the defect in RXL substrate phosphorylation, because phosphorylation can be rescued by restoring a cyclin A-substrate interaction in a heterologous manner. In addition, the conserved hydrophobic patch is important for cyclin A function in cells, contributing to cyclin A's ability to drive cells out of the G1 phase of the cell cycle. Thus, we define a mechanism by which cyclins can recruit substrates to cdks, and our results support the notion that a high local concentration of substrate provided by a protein-protein interaction distant from the active site is critical for phosphorylation by cdks. PMID- 9724725 TI - Glucocorticoids and progestins signal the initiation and enhance the rate of myelin formation. AB - Dexamethasone and progesterone have been found to accelerate the time of initiation and enhance the rate of myelin synthesis in Schwann cell/neuronal cocultures. The expression of mRNA for cytochrome P450scc (converts cholesterol to pregnenolone), 3beta-hydroxysteroid dehydrogenase (converts pregnenolone to progesterone), and the progesterone receptor were detected and markedly induced during peak myelin formation in the cocultures. The mRNA for the glucocorticoid receptor was detected, but was found to be constituitively expressed. In addition, the specific activity of 3beta-hydroxysteroid dehydrogenase was measured and found to increase by 10-fold. The mRNA for cytochrome P450scc and 3beta-hydroxysteroid dehydrogenase also were found to be induced during the differentiation of O-2A precursor cells to oligodendrocytes. Fibroblast growth factor and platelet-derived growth factor were found to have proliferative effects on Schwann cells, but they had no effect on the initiation or the rate of myelin formation. These results demonstrate that myelin-forming cells have inducible enzymes responsible for steroid biosynthesis and suggest a critical role for endogenous steroid hormones in signaling the initiation and enhancing the rate of myelin formation. PMID- 9724728 TI - Branch migration during Rad51-promoted strand exchange proceeds in either direction. AB - The Saccharomyces cerevisiae Rad51 protein is important for genetic recombination and repair of DNA double-strand breaks in vivo and can promote strand exchange between linear double-stranded DNA and circular single-stranded DNA in vitro. However, unlike Escherichia coli RecA, Rad51 requires an overhanging complementary 3' or 5' end to initiate strand exchange; given that fact, we previously surmised that the fully exchanged molecules resulted from branch migration in either direction depending on which type of end initiated the joint molecule. Our present experiments confirm that branch migration proceeds in either direction, the polarity depending on whether a 3' or 5' end initiates the joint molecules. Furthermore, heteroduplex DNA is formed rapidly, first at the overhanging end of the linear double-stranded DNA's complementary strand and then more slowly by progressive lengthening of the heteroduplex region until strand exchange is complete. Although joint molecule formation occurs equally efficiently when initiated with a 3' or 5' overhanging end, branch migration proceeds more rapidly when it is initiated by an overhanging 3' end, i.e., in the 5' to 3' direction with respect to the single-stranded DNA. PMID- 9724726 TI - Design, synthesis, and characterization of a photoactivatable flavocytochrome molecular maquette. AB - We report the construction of a synthetic flavo-heme protein that incorporates two major physiological activities of flavoproteins: light activation of flavin analogous to DNA photolyase and rapid intramolecular electron transfer between the flavin and heme cofactors as in several oxidoreductases. The functional tetra alpha-helix protein comprises two 62-aa helix-loop-helix subunits. Each subunit contains a single cysteine to which flavin (7-acetyl-10-methylisoalloxazine) is covalently attached and two histidines appropriately positioned for bis-his coordination of heme cofactors. Both flavins and hemes are situated within the hydrophobic core of the protein. Intramolecular electron transfer from flavosemiquinone generated by photoreduction from a sacrificial electron donor in solution was examined between protoporphyrin IX and 1-methyl-2-oxomesoheme XIII. Laser pulse-activated electron transfer from flavin to meso heme occurs on a 100 ns time scale, with a favorable free energy of approximately -100 meV. Electron transfer from flavin to the lower potential protoporphyrin IX, with an unfavorable free energy, can be induced after a lag phase under continuous light illumination. Thus, the supporting peptide matrix provides an excellent framework for the positioning of closely juxtaposed redox groups capable of facilitating intramolecular electron transfer and begins to clarify in a simplified and malleable system the natural engineering of flavoproteins. PMID- 9724727 TI - Expression of mouse telomerase catalytic subunit in embryos and adult tissues. AB - Telomerase is a ribonucleoprotein complex that elongates telomeres, allowing the stable maintenance of chromosomes during multiple cell divisions. Here, we describe the isolation and characterization of the catalytic subunit of mouse telomerase, mTERT (mouse telomerase reverse transcriptase), an essential protein component of the telomerase complex. During embryonic development, mTERT mRNA is abundantly expressed in the whole embryo, especially in regions of intense proliferation. We found that the mTERT mRNA expression in both embryonic and adult tissues is independent of the essential RNA component of telomerase, mTR, and therefore, of the formation of active telomerase complexes. mTERT protein is present exclusively in tissues with telomerase activity, such as testis, spleen, and thymus. mTERT protein is barely detectable in the thymus of mTR-/- mice, suggesting that mTERT protein stability in this tissue may depend on the actual assembly of active telomerase complexes. Finally, we found that mouse and human telomerase catalytic subunit is located in the cell nucleus, and its localization is not regulated during cell cycle progression. PMID- 9724729 TI - Cloning and characterization of a third human lysyl hydroxylase isoform. AB - Lysyl hydroxylase (EC 1.14.11.4), a homodimer, catalyzes the formation of hydroxylysine in collagens. Recently, an isoenzyme termed lysyl hydroxylase 2 has been cloned from human sources [M. Valtavaara, H. Papponen, A.-M. Pirttila, K. Hiltunen, H. Helander and R. Myllyla (1997) J. Biol. Chem. 272, 6831-6834]. We report here on the cloning of a third human lysyl hydroxylase isoenzyme, termed lysyl hydroxylase 3. The cDNA clones encode a 738 amino acid polypeptide, including a signal peptide of 24 residues. The overall amino acid sequence identity between the processed human lysyl hydroxylase 3 and 1 polypeptides is 59%, and that between the processed lysyl hydroxylase 3 and 2 polypeptides is 57%, whereas the identity to the processed Caenorhabditis elegans polypeptide is only 45%. All four recently identified critical residues at the catalytic site, two histidines, one aspartate, and one arginine, are conserved in all these polypeptides. The mRNA for lysyl hydroxylase 3 was found to be expressed in a variety of tissues, but distinct differences appear to exist in the expression patterns of the three isoenzyme mRNAs. Recombinant lysyl hydroxylase 3 expressed in insect cells by means of a baculovirus vector was found to be more soluble than lysyl hydroxylase 1 expressed in the same cell type. No differences in catalytic properties were found between the recombinant lysyl hydroxylase 3 and 1 isoenzymes. Deficiency in lysyl hydroxylase 1 activity is known to cause the type VI variant of the Ehlers-Danlos syndrome, and it is therefore possible that deficiency in lysyl hydroxylase 3 activity may lead to some other variant of this syndrome or to some other heritable connective tissue disorder. PMID- 9724730 TI - Evolution of a quadripartite hybrid virus by interspecific exchange and recombination between replicase components of two related tripartite RNA viruses. AB - Cucumber mosaic virus (CMV) and tomato aspermy virus (TAV) belong to the Cucumovirus genus. They have a tripartite genome consisting of single-stranded RNAs, designated 1, 2, and 3. Previous studies have shown that viable pseudorecombinants could be created in vitro by reciprocal exchanges between CMV and TAV RNA 3, but exchanges of RNAs 1 and 2 were replication deficient. When we coinoculated CMV RNAs 2 and 3 along with TAV RNAs 1 and 2 onto Nicotiana benthamiana, a hybrid quadripartite virus appeared that consisted of TAV RNA 1, CMV RNAs 2 and 3, and a distinctive chimeric RNA originating from a recombination between CMV RNA 2 and the 3'-terminal 320 nucleotides of TAV RNA 2. This hybrid arose by means of segment reassortment and RNA recombination to produce an interspecific hybrid with the TAV helicase subunit and the CMV polymerase subunit. To our knowledge, this is the first report demonstrating the evolution of a new plant or animal virus strain containing an interspecific hybrid replicase complex. PMID- 9724731 TI - The three members of the pocket proteins family share the ability to repress E2F activity through recruitment of a histone deacetylase. AB - The transcription factor E2F plays a major role in cell cycle control in mammalian cells. E2F binding sites, which are present in the promoters of a variety of genes required for S phase, shift from a negative to a positive role in transcription at the commitment point, a crucial point in G1 that precedes the G1/S transition. Before the commitment point, E2F activity is repressed by members of the pocket proteins family. This repression is believed to be crucial for the proper control of cell growth. We have previously shown that Rb, the founding member of the pocket proteins family, represses E2F1 activity by recruiting the histone deacetylase HDAC1. Here, we show that the two other members of the pocket proteins family, p107 and p130, also are able to interact physically with HDAC1 in live cells. HDAC1 interacts with p107 and Rb through an "LXCXE"-like motif, similar to that used by viral transforming proteins to bind and inactivate pocket proteins. Indeed, we find that the viral transforming protein E1A competes with HDAC1 for p107 interaction. We also demonstrate that p107 is able to interact simultaneously with HDAC1 and E2F4, suggesting a model in which p107 recruits HDAC1 to repress E2F sites. Indeed, we demonstrate that histone deacetylase activity is involved in the p107- or p130-induced repression of E2F4. Taken together, our data suggest that all members of the E2F family are regulated in early G1 by similar complexes, containing a pocket protein and the histone deacetylase HDAC1. PMID- 9724733 TI - Lagging strand replication of rolling-circle plasmids: specific recognition of the ssoA-type origins in different gram-positive bacteria. AB - Many bacterial plasmids replicate by a rolling-circle mechanism that involves the generation of single-stranded DNA (ssDNA) intermediates. Replication of the lagging strand of such plasmids initiates from their single strand origin (sso). Many different types of ssos have been identified. One group of ssos, termed ssoA, which have conserved sequence and structural features, function efficiently only in their natural hosts in vivo. To study the host specificity of sso sequences, we have analyzed the functions of two closely related ssoAs belonging to the staphylococcal plasmid pE194 and the streptococcal plasmid pLS1 in Staphylococcus aureus. The pLS1 ssoA functioned poorly in vivo in S. aureus as evidenced by accumulation of high levels of ssDNA but supported efficient replication in vitro in staphylococcal extracts. These results suggest that one or more host factors that are present in sufficient quantities in S. aureus cell free extracts may be limiting in vivo. Mapping of the initiation points of lagging strand synthesis in vivo and in vitro showed that DNA synthesis initiates from specific sites within the pLS1 ssoA. These results demonstrate that specific initiation of replication can occur from the pLS1 ssoA in S. aureus although it plays a minimal role in lagging strand synthesis in vivo. Therefore, the poor functionality of the pLS1 in vivo in a nonnative host is caused by the low efficiency rather than a lack of specificity of the initiation process. We also have identified ssDNA promoters and mapped the primer RNAs synthesized by the S. aureus and Bacillus subtilis RNA polymerases from the pE194 and pLS1 ssoAs. The S. aureus RNA polymerase bound more efficiently to the native pE194 ssoA as compared with the pLS1 ssoA, suggesting that the strength of RNA polymerase-ssoA interaction may play a major role in the functionality of the ssoA sequences in Gram-positive bacteria. PMID- 9724732 TI - Farnesyltransferase inhibitors induce dramatic morphological changes of KNRK cells that are blocked by microtubule interfering agents. AB - Farnesyltransferase inhibitors (FTIs) exhibit the remarkable ability to inhibit transformed phenotypes of a variety of human cancer cell lines and to block the growth of cancer cells in a number of animal model systems. In this paper, we report that the addition of FTI to v-K-ras- transformed NRK cells (KNRK) results in dramatic morphological changes. Within 24 h after the addition of FTI, the round morphology of KNRK cells was changed to an elongated (flattened and spread out) morphology resembling those of untransformed NRK cells. No morphological effects were seen when similar concentrations of FTI were added to NRK cells. Phalloidin staining showed that FTI treatment did not restore the disrupted actin cytoskeleton in KNRK cells. In contrast, FTI addition resulted in the appearance of extensive microtubule networks in KNRK cells. The addition of a low concentration (1.2 nM) of vincristine or vinblastine, agents that interfere with microtubule dynamics, blocked the FTI-induced morphological changes in KNRK cells. In contrast, cytochalasin B, which interferes with actin polymerization, did not block the morphological changes. The FTI-induced morphological changes were associated with a decrease in the percentage of cells in S-phase, and the addition of 1.2 nM vincristine did not have additional effects on cell cycle progression. A higher concentration (12 nM) of vincristine caused synergistic effect with FTI to enrich dramatically KNRK cells in G2/M phase. These results suggest that FTI affects cell morphology and that microtubule dynamics are involved in these processes. PMID- 9724734 TI - The location of the carboxy-terminal region of gamma chains in fibrinogen and fibrin D domains. AB - Elongated fibrinogen molecules are comprised of two outer "D" domains, each connected through a "coiled-coil" region to the central "E" domain. Fibrin forms following thrombin cleavage in the E domain and then undergoes intermolecular end to-middle D:E domain associations that result in double-stranded fibrils. Factor XIIIa mediates crosslinking of the C-terminal regions of gamma chains in each D domain (the gammaXL site) by incorporating intermolecular epsilon-(gamma glutamyl)lysine bonds between amine donor gamma406 lysine of one gamma chain and a glutamine acceptor at gamma398 or gamma399 of another. Several lines of evidence show that crosslinked gamma chains extend "transversely" between the strands of each fibril, but other data suggest instead that crosslinked gamma chains can only traverse end-to-end-aligned D domains within each strand. To examine this issue and determine the location of the gammaXL site in fibrinogen and assembled fibrin fibrils, we incorporated an amine donor, thioacetyl cadaverine, into glutamine acceptor sites in fibrinogen in the presence of XIIIa, and then labeled the thiol with a relatively small (0.8 nm diameter) electron dense gold cluster compound, undecagold monoaminopropyl maleimide (Au11). Fibrinogen was examined by scanning transmission electron microscopy to locate Au11-cadaverine-labeled gamma398/399 D domain sites. Seventy-nine percent of D domain Au11 clusters were situated in middle to proximal positions relative to the end of the molecule, with the remaining Au11 clusters in a distal position. In fibrin fibrils, D domain Au11 clusters were located in middle to proximal positions. These findings show that most C-terminal gamma chains in fibrinogen or fibrin are oriented toward the central domain and indicate that gammaXL sites in fibrils are situated predominantly between strands, suitably aligned for transverse crosslinking. PMID- 9724735 TI - A potential mediator of collagenous block copolymer gradients in mussel byssal threads. AB - Mussel byssal threads contain unusual block copolymer-like proteins that combine collagen with flanking domains that resemble silk-fibroin (preCol-D) or elastin (preCol-P). These are distributed in complementary gradients along the length of the threads and as precursors in the mussel foot. We discuss a 76-kDa precursor, preCol-NG, from a cDNA library of the foot where it has no gradient but rather is distributed evenly along the distal to proximal axis. A pepsin-resistant fragment of preCol-NG has been confirmed in byssal threads. Like preCol-D and -P, this protein has a central collagenous domain, flanking domains, an acidic patch, and histidine-rich termini. The flanking domains of preCol-NG resemble the glycine rich proteins of plant cell walls with tandem XGlyn repeats where X denotes alanine, leucine, or asparagine but not proline. Similarity with the (glycine alanine) repeats and poly(alanine) runs of arthropod silks also exists. Based on available evidence, a model of preCol axial assembly is proposed in which preCol NG functions as a mediator between preCol-D/-P molecules. This is consistent with the observed progression of mechanical properties in byssal threads. PMID- 9724736 TI - A method for directed evolution and functional cloning of enzymes. AB - A general scheme is described for the in vitro evolution of protein catalysts in a biologically amplifiable system. Substrate is covalently and site specifically attached by a flexible tether to the pIII coat protein of a filamentous phage that also displays the catalyst. Intramolecular conversion of substrate to product provides a basis for selecting active catalysts from a library of mutants, either by release from or attachment to a solid support. This methodology has been developed with the enzyme staphylococcal nuclease as a model. An analysis of factors influencing the selection efficiency is presented, and it is shown that phage displaying staphylococcal nuclease can be enriched 100 fold in a single step from a library-like ensemble of phage displaying noncatalytic proteins. Additionally, this approach should allow one to functionally clone natural enzymes, based on their ability to catalyze specific reactions (e.g., glycosyl transfer, sequence-specific proteolysis or phosphorylation, polymerization, etc.) rather than their sequence- or structural homology to known enzymes. PMID- 9724737 TI - Multiple promoters direct the tissue-specific expression of novel N-terminal variant human vitamin D receptor gene transcripts. AB - The effects of 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] are mediated by the vitamin D receptor (VDR), a member of the nuclear receptor superfamily of transcriptional regulators. We have identified upstream exons of the human (h) VDR gene that are incorporated into variant transcripts, two of which encode N terminal variant receptor proteins. Expression of the hVDR gene, which spans more than 60 kb and consists of at least 14 exons, is directed by two distinct promoters. A tissue-specific distal promoter generates unique transcripts in tissues involved in calcium regulation by 1, 25-(OH)2D3 and can direct the expression of a luciferase reporter gene in a cell line-specific manner. These major N-terminal differences in hVDR transcripts, potentially resulting in structural differences in the expressed receptor, may contribute to cellular responsiveness to 1,25-(OH)2D3 through tissue differences in the regulation of VDR expression. PMID- 9724738 TI - Footprints on the viral DNA ends in moloney murine leukemia virus preintegration complexes reflect a specific association with integrase. AB - Retroviral DNA integration is mediated by the preintegration complex, a large nucleoprotein complex derived from the core of the infecting virion. We previously have used Mu-mediated PCR to probe the nucleoprotein organization of Moloney murine leukemia virus preintegration complexes. A region of protection spans several hundred base pairs at each end of the viral DNA, and strong enhancements are present near the termini. Here, we show that these footprints reflect a specific association between integrase and the viral DNA ends in functional preintegration complexes. Barrier-to-autointegration factor, a cellular protein that blocks autointegration of Moloney murine leukemia virus DNA, also plays an indirect role in generating the footprints at the ends of the viral DNA. We have exploited Mu-mediated PCR to examine the effect of mutations at the viral DNA termini on complex formation. We find that a replication competent mutant with a deletion at one end of the viral DNA still exhibits a strong enhancement about 20 bp from the terminus of the mutant DNA end. The site of the enhancement therefore appears to be at a fixed distance from the ends of the viral DNA. We also find that a mutation at one end of the viral DNA, which renders the virus incompetent for replication, abolishes the enhancements and protection at both the U3 and U5 ends. A pair of functional viral DNA ends therefore are required to interact before the chemical step of 3' end processing. PMID- 9724739 TI - MEKK1/JNK signaling stabilizes and activates p53. AB - Activation of the tumor suppressor p53 by stress and damage stimuli often correlates with induction of stress kinases, Jun-NH2 kinase (JNK). As JNK association with p53 plays an important role in p53 stability, in the present study we have elucidated the relationship between the JNK-signaling pathway and p53 stability and activity. Expression of a constitutively active form of JNKK upstream kinase, mitogen-activated protein kinase kinase kinase (DeltaMEKK1), increased the level of the exogenously transfected form of p53 in p53 null (10.1) cells as well as of endogenous p53 in MCF7 breast cancer cells. Increased p53 level by forced expression of DeltaMEKK1 coincided with a decrease in p53 ubiquitination in vivo and with prolonged p53 half-life. Computerized modeling of the JNK-binding site (amino acids 97-116; p7 region) enabled us to design mutations of exposed residues within this region. Respective mutations (p53(101-5 8)) and deletion (p53(Deltap7)) forms of p53 did not exhibit the same increase in p53 levels upon DeltaMEKK1 expression. In vitro phosphorylation of p53 by JNK abolished Mdm2 binding and targeting of p53 ubiquitination. Similarly, DeltaMEKK1 expression increased p53 phosphorylation by immunopurified JNK and dissociated p53-Mdm2 complexes. Transcriptional activity of p53, as measured via mdm2 promoter-driven luciferase, exhibited a substantial increase in DeltaMEKK1 expressing cells. Cotransfection of p53 and DeltaMEKK1 into p53 null cells potentiated p53-dependent apoptosis, suggesting that MEKK1 effectors contribute to the ability of p53 to mediate programmed cell death. Our results point to the role of MEKK1-JNK signaling in p53 stability, transcriptional activities, and apoptotic capacity as part of the cellular response to stress. PMID- 9724740 TI - Implications of macromolecular crowding for signal transduction and metabolite channeling. AB - The effect of different total enzyme concentrations on the flux through the bacterial phosphoenolpyruvate:carbohydrate phosphotransferase system (PTS) in vitro was determined by measuring PTS-mediated carbohydrate phosphorylation at different dilutions of cell-free extract of Escherichia coli. The dependence of the flux on the protein concentration was more than linear but less than quadratic. The combined flux-response coefficient of the four enzymes constituting the glucose PTS decreased slightly from values of approximately 1.8 with increasing protein concentrations in the assay. Addition of the macromolecular crowding agents polyethylene glycol (PEG) 6000 and PEG 35000 led to a sharper decrease in the combined flux-response coefficient, in one case to values of approximately 1. PEG 6000 stimulated the PTS flux at lower protein concentrations and inhibited the flux at higher protein concentrations, with the transition depending on the PEG 6000 concentration. This suggests that macromolecular crowding decreases the dissociation rate constants of enzyme complexes. High concentrations of the microsolute glycerol did not affect the combined flux-response coefficient. The data could be explained with a kinetic model of macromolecular crowding in a two-enzyme group-transfer pathway. Our results suggest that, because of the crowded environment in the cell, the different PTS enzymes form complexes that live long on the time-scale of their turnover. The implications for the metabolic behavior and control properties of the PTS, and for the effect of macromolecular crowding on nonequilibrium processes, are discussed. PMID- 9724741 TI - The domain structure of protoporphyrinogen oxidase, the molecular target of diphenyl ether-type herbicides. AB - Protoporphyrinogen oxidase (EC 1-3-3-4), the 60-kDa membrane-bound flavoenzyme that catalyzes the final reaction of the common branch of the heme and chlorophyll biosynthesis pathways in plants, is the molecular target of diphenyl ether-type herbicides. It is highly resistant to proteases (trypsin, endoproteinase Glu-C, or carboxypeptidases A, B, and Y), because the protein is folded into an extremely compact form. Trypsin maps of the native purified and membrane-bound yeast protoporphyrinogen oxidase show that this basic enzyme (pI > 8.5) was cleaved at a single site under nondenaturing conditions, generating two peptides with relative molecular masses of 30,000 and 35,000. The endoproteinase Glu-C also cleaved the protein into two peptides with similar masses, and there was no additional cleavage site under mild denaturing conditions. N-terminal peptide sequence analysis of the proteolytic (trypsin and endoproteinase Glu-C) peptides showed that both cleavage sites were located in putative connecting loop between the N-terminal domain (25 kDa) with the betaalphabeta ADP-binding fold and the C-terminal domain (35 kDa), which possibly is involved in the binding of the isoalloxazine moiety of the FAD cofactor. The peptides remained strongly associated and fully active with the Km for protoporphyrinogen and the Ki for various inhibitors, diphenyl-ethers, or diphenyleneiodonium derivatives, identical to those measured for the native enzyme. However, the enzyme activity of the peptides was much more susceptible to thermal denaturation than that of the native protein. Only the C-terminal domain of protoporphyrinogen oxidase was labeled specifically in active site-directed photoaffinity-labeling experiments. Trypsin may have caused intramolecular transfer of the labeled group to reactive components of the N-terminal domain, resulting in nonspecific labeling. We suggest that the active site of protoporphyrinogen oxidase is in the C-terminal domain of the protein, at the interface between the C- and N-terminal domains. PMID- 9724742 TI - Activation of iron regulatory protein-1 by oxidative stress in vitro. AB - Iron regulatory protein-1 (IRP-1), a central cytoplasmic regulator of cellular iron metabolism, is rapidly activated by oxidative stress to bind to mRNA iron responsive elements. We have reconstituted the response of IRP-1 to extracellular H2O2 in a system derived from murine B6 fibroblasts permeabilized with streptolysin-O. This procedure allows separation of the cytosol from the remainder of the cells (cell pellet). IRP-1 in the cytosolic fraction fails to be directly activated by addition of H2O2. IRP-1 activation requires the presence of a nonsoluble, possibly membrane-associated component in the cell pellet. The streptolysin-O-based in vitro system faithfully recapitulates characteristic hallmarks of IRP-1 activation by H2O2 in intact cells. We show that the H2O2 mediated activation of IRP-1 is temperature dependent and sensitive to treatment with calf intestinal alkaline phosphatase (CIAP). Although IRP-1 activation is unaffected by addition of excess ATP or GTP to this in vitro system, it is negatively affected by the nonhydrolyzable nucleotide analogs adenylyl imidodiphosphate and guanylyl-imidophosphate and completely blocked by ATP-gammaS and GTP-gammaS. The in vitro reconstitution of this oxidative stress-induced pathway has opened a different avenue for the biochemical dissection of the regulation of mammalian iron metabolism by oxidative stress. Our data show that H2O2 must be sensed to stimulate a pathway to activate IRP-1. PMID- 9724743 TI - Structure of FokI has implications for DNA cleavage. AB - FokI is a member an unusual class of restriction enzymes that recognize a specific DNA sequence and cleave nonspecifically a short distance away from that sequence. FokI consists of an N-terminal DNA recognition domain and a C-terminal cleavage domain. The bipartite nature of FokI has led to the development of artificial enzymes with novel specificities. We have solved the structure of FokI to 2.3 A resolution. The structure reveals a dimer, in which the dimerization interface is mediated by the cleavage domain. Each monomer has an overall conformation similar to that found in the FokI-DNA complex, with the cleavage domain packing alongside the DNA recognition domain. In corroboration with the cleavage data presented in the accompanying paper in this issue of Proceedings, we propose a model for FokI DNA cleavage that requires the dimerization of FokI on DNA to cleave both DNA strands. PMID- 9724744 TI - FokI dimerization is required for DNA cleavage. AB - FokI is a type IIs restriction endonuclease comprised of a DNA recognition domain and a catalytic domain. The structural similarity of the FokI catalytic domain to the type II restriction endonuclease BamHI monomer suggested that the FokI catalytic domains may dimerize. In addition, the FokI structure, presented in an accompanying paper in this issue of Proceedings, reveals a dimerization interface between catalytic domains. We provide evidence here that FokI catalytic domain must dimerize for DNA cleavage to occur. First, we show that the rate of DNA cleavage catalyzed by various concentrations of FokI are not directly proportional to the protein concentration, suggesting a cooperative effect for DNA cleavage. Second, we constructed a FokI variant, FokN13Y, which is unable to bind the FokI recognition sequence but when mixed with wild-type FokI increases the rate of DNA cleavage. Additionally, the FokI catalytic domain that lacks the DNA binding domain was shown to increase the rate of wild-type FokI cleavage of DNA. We also constructed an FokI variant, FokD483A, R487A, which should be defective for dimerization because the altered residues reside at the putative dimerization interface. Consistent with the FokI dimerization model, the variant FokD483A, R487A revealed greatly impaired DNA cleavage. Based on our work and previous reports, we discuss a pathway of DNA binding, dimerization, and cleavage by FokI endonuclease. PMID- 9724745 TI - Epidermal trans-urocanic acid and the UV-A-induced photoaging of the skin. AB - The premature photoaging of the skin is mediated by the sensitization of reactive oxygen species after absorption of ultraviolet radiation by endogenous chromophores. Yet identification of UV-A-absorbing chromophores in the skin that quantitatively account for the action spectra of the physiological responses of photoaging has remained elusive. This paper reports that the in vitro action spectrum for singlet oxygen generation after excitation of trans-urocanic acid mimics the in vivo UV-A action spectrum for the photosagging of mouse skin. The data presented provide evidence suggesting that the UV-A excitation of trans urocanic acid initiates chemical processes that result in the photoaging of skin. PMID- 9724746 TI - Homologous pairing in stretched supercoiled DNA. AB - By using elastic measurements on single DNA molecules, we show that stretching a negatively supercoiled DNA activates homologous pairing in physiological conditions. These experiments indicate that a stretched unwound DNA locally denatures to alleviate the force-driven increase in torsional stress. This is detected by hybridization with 1 kb of homologous single-stranded DNA probes. The stretching force involved (approximately 2 pN) is small compared with those typically developed by molecular motors, suggesting that this process may be relevant to DNA processing in vivo. We used this technique to monitor the progressive denaturation of DNA as it is unwound and found that distinct, stable denaturation bubbles formed, beginning in A+T-rich regions. PMID- 9724747 TI - The ATPase and protease domains of yeast mitochondrial Lon: roles in proteolysis and respiration-dependent growth. AB - The ATP-dependent Lon protease of Saccharomyces cerevisiae mitochondria is required for selective proteolysis in the matrix, maintenance of mitochondrial DNA, and respiration-dependent growth. Lon may also possess a chaperone-like function that facilitates protein degradation and protein-complex assembly. To understand the influence of Lon's ATPase and protease activities on these functions, we examined several Lon mutants for their ability to complement defects of Lon-deleted yeast cells. We also developed a rapid procedure for purifying yeast Lon to homogeneity to study the enzyme's activities and oligomeric state. A point mutation in either the ATPase or the protease site strongly inhibited the corresponding activity of the pure protein but did not alter the protein's oligomerization; when expressed at normal low levels neither of these mutant enzymes supported respiration-dependent growth of Lon-deleted cells. When the ATPase- or the protease-containing regions of Lon were expressed as separate truncated proteins, neither could support respiration-dependent growth of Lon-deleted cells; however, coexpression of these two separated regions sustained wild-type growth. These results suggest that yeast Lon contains two catalytic domains that can interact with one another even as separate proteins, and that both are essential for the different functions of Lon. PMID- 9724748 TI - Suppression of adenovirus E1A-induced apoptosis by mutated p53 is overcome by coexpression with Id proteins. AB - The rat 3Y1 derivative cell lines, EId10 and EId23, established by introducing the adenovirus E1A12S, Id-1H, and Id-2H cDNAs linked to the hormone-inducible promoter, express these proteins upon treatment with dexamethasone and elicit apoptosis, although these cell lines express mutated p53. The E1A mutants containing a deletion in either the N terminus or the conserved region 1 were unable to induce apoptosis in cooperation with Ids. Western blot analysis of the immunoprecipitates prepared from the dexamethasone-treated EId10 cell extract showed that Id-2H preferentially binds to E1A and E2A (E12/E47) helix-loop-helix transcription factors in vivo, but scarcely to the retinoblastoma protein. After induction of E1A and Ids, EId10 and EId23 cells began to accumulate in S phase and undergo apoptosis before entering G2 phase, suggesting that abnormal synthesis of DNA induced by coexpression of E1A, Id-1H, and Id-2H results in the induction of apoptosis. Apoptosis also is induced in mouse A40 (p53-/-) cells by E1A alone or E1A plus Ids after transient transfection of the expression vectors. The induction of apoptosis is stimulated by coexpression with wild-type p53; however, apoptosis induced by E1A alone was suppressed completely by coexpression with mutated p53, whereas apoptosis induced by E1A plus Ids was stimulated by the mutated p53 as done by wild-type p53. These results suggest that the suppressive function of mutated p53 is overcome by Ids. PMID- 9724749 TI - The adenomatous polyposis coli-binding protein EB1 is associated with cytoplasmic and spindle microtubules. AB - The evolutionarily conserved protein EB1 originally was identified by its physical association with the carboxyl-terminal portion of the adenomatous polyposis coli (APC) tumor suppressor protein, an APC domain commonly mutated in familial and sporadic forms of colorectal neoplasia. The subcellular localization of EB1 in epithelial cells was studied by using immunofluorescence and biochemical techniques. EB1 colocalized both to cytoplasmic microtubules in interphase cells and to spindle microtubules during mitosis, with pronounced centrosome staining. The cytoskeletal array detected by anti-EB1 antibody was abolished by incubation of the cells with nocodazole, an agent that disrupts microtubules; upon drug removal, EB1 localized to the microtubule-organizing center. Immunofluorescence analysis of SW480, a colon cancer cell line that expresses only carboxyl-terminal-deleted APC unable to interact with EB1, demonstrated that EB1 remained localized to the microtubule cytoskeleton, suggesting that this pattern of subcellular distribution is not mediated by its interaction with APC. In vitro cosedimentation with taxol-stabilized microtubules demonstrated that a significant fraction of EB1 associated with microtubules. Recent studies of the yeast EB1 homologues Mal3 and Bim1p have demonstrated that both proteins localize to microtubules and are important in vivo for microtubule function. Our results demonstrate that EB1 is a novel component of the microtubule cytoskeleton in mammalian cells. Associating with the mitotic apparatus, EB1 may play a physiologic role connecting APC to cellular division, coordinating the control of normal growth and differentiation processes in the colonic epithelium. PMID- 9724750 TI - WW domain-mediated interactions reveal a spliceosome-associated protein that binds a third class of proline-rich motif: the proline glycine and methionine rich motif. AB - Pre-mRNA splicing requires the bridging of the 5' and 3' ends of the intron. In yeast, this bridging involves interactions between the WW domains in the splicing factor PRP40 and a proline-rich domain in the branchpoint binding protein, BBP. Using a proline-rich domain derived from formin (a product of the murine limb deformity locus), we have identified a family of murine formin binding proteins (FBP's), each of which contains one or more of a special class of tyrosine-rich WW domains. Two of these WW domains, in the proteins FBP11 and FBP21, are strikingly similar to those found in the yeast splicing factor PRP40. We show that FBP21 is present in highly purified spliceosomal complex A, is associated with U2 snRNPs, and colocalizes with splicing factors in nuclear speckle domains. Moreover, FBP21 interacts directly with the U1 snRNP protein U1C, the core snRNP proteins SmB and SmB', and the branchpoint binding protein SF1/mBBP. Thus, FBP21 may play a role in cross-intron bridging of U1 and U2 snRNPs in the mammalian A complex. PMID- 9724751 TI - Reconstitution of HIV-1 rev nuclear export: independent requirements for nuclear import and export. AB - The Rev protein of HIV-1 actively shuttles between nucleus and cytoplasm and mediates the export of unspliced retroviral RNAs. The localization of shuttling proteins such as Rev is controlled by the relative rates of nuclear import and export. To study nuclear export in isolation, we generated cell lines expressing a green fluorescent protein-labeled chimeric protein consisting of HIV-1 Rev and a hormone-inducible nuclear localization sequence. Steroid removal switches off import thus allowing direct visualization of the Rev export pathway in living cells. After digitonin permeabilization of these cells, we found that a functional nuclear export sequence (NES), ATP, and fractionated cytosol were sufficient for nuclear export in vitro. Nuclear pore-specific lectins and leptomycin B were potent export inhibitors. Nuclear export was not inhibited by antagonists of calcium metabolism that block nuclear import. These data further suggest that nuclear pores do not functionally close when luminal calcium stores are depleted. The distinct requirements for nuclear import and export argue that these competing processes may be regulated independently. This system should have wide applicability for the analysis of nuclear import and export. PMID- 9724752 TI - Relationship between donor age and the replicative lifespan of human cells in culture: a reevaluation. AB - Normal human diploid fibroblasts have a finite replicative lifespan in vitro, which has been postulated to be a cellular manifestation of aging in vivo. Several studies have shown an inverse relationship between donor age and fibroblast culture replicative lifespan; however, in all cases, the correlation was weak, and, with few exceptions, the health status of the donors was unknown. We have determined the replicative lifespans of 124 skin fibroblast cell lines established from donors of different ages as part of the Baltimore Longitudinal Study of Aging. All of the donors were medically examined and were declared "healthy," according to Baltimore Longitudinal Study of Aging protocols, at the time the biopsies were taken. Both long- and short-lived cell lines were observed in all age groups, but no significant correlation between the proliferative potential of the cell lines and donor age was found. A comparison of multiple cell lines established from the same donors at different ages also failed to reveal any significant trends between proliferative potential and donor age. The rate of [3H]thymidine incorporation and the initial rates of growth during the first few subcultivations were examined in a subset of cell lines and were found to be significantly greater in fetal lines than in postnatal lines. Cell lines established from adults did not vary significantly either in initial growth rate or in [3H]thymidine incorporation. These results clearly indicate that, if health status and biopsy conditions are controlled, the replicative lifespan of fibroblasts in culture does not correlate with donor age. PMID- 9724753 TI - Regulation of sorting and post-Golgi trafficking of rhodopsin by its C-terminal sequence QVS(A)PA. AB - Several mutations that cause severe forms of the human disease autosomal dominant retinitis pigmentosa cluster in the C-terminal region of rhodopsin. Recent studies have implicated the C-terminal domain of rhodopsin in its trafficking on specialized post-Golgi membranes to the rod outer segment of the photoreceptor cell. Here we used synthetic peptides as competitive inhibitors of rhodopsin trafficking in the frog retinal cell-free system to delineate the potential regulatory sequence within the C terminus of rhodopsin and model the effects of severe retinitis pigmentosa alleles on rhodopsin sorting. The rhodopsin C terminal sequence QVS(A)PA is highly conserved among different species. Peptides that correspond to the C terminus of bovine (amino acids 324-348) and frog (amino acids 330-354) rhodopsin inhibited post-Golgi trafficking by 50% and 60%, respectively, and arrested newly synthesized rhodopsin in the trans-Golgi network. Peptides corresponding to the cytoplasmic loops of rhodopsin and other control peptides had no effect. When three naturally occurring mutations: Q344ter (lacking the last five amino acids QVAPA), V345M, and P347S were introduced into the frog C-terminal peptide, the inhibitory activity of the peptides was no longer detectable. These observations suggest that the amino acids QVS(A)PA comprise a signal that is recognized by specific factors in the trans-Golgi network. A lack of recognition of this sequence, because of mutations in the last five amino acids causing autosomal dominant retinitis pigmentosa, most likely results in abnormal post-Golgi membrane formation and in an aberrant subcellular localization of rhodopsin. PMID- 9724755 TI - MLL, a mammalian trithorax-group gene, functions as a transcriptional maintenance factor in morphogenesis. AB - Determinative events in vertebrate embryogenesis appear to require the continuous expression of spatial regulators such as the clustered homeobox genes. The mechanisms that govern long-term patterns of gene expression are not well understood. In Drosophila, active and silent states of developmentally regulated loci are maintained by trithorax and Polycomb group. We have examined the developmental role of a mammalian homolog of trx and putative oncogene, Mll. Knockout mice reveal that Mll is required for maintenance of gene expression early in embryogenesis. Downstream targets of Mll including Hoxa7 are activated appropriately in the absence of Mll but require Mll for sustaining their expression. The Mll-/- phenotype manifests later in development and is characterized by branchial arch dysplasia and aberrant segmental boundaries of spinal ganglia and somites. Thus, Mll represents an essential mechanism of transcriptional maintenance in mammalian development, which functions in multiple morphogenetic processes. PMID- 9724754 TI - Inhibition of Stat1-mediated gene activation by PIAS1. AB - STAT (signal transducer and activator of transcription) proteins are latent cytoplasmic transcription factors that become activated by tyrosine phosphorylation in response to cytokine stimulation. Tyrosine phosphorylated STATs dimerize and translocate into the nucleus to activate specific genes. Different members of the STAT protein family have distinct functions in cytokine signaling. Biochemical and genetic analysis has demonstrated that Stat1 is essential for gene activation in response to interferon stimulation. Although progress has been made toward understanding STAT activation, little is known about how STAT signals are down-regulated. We report here the isolation of a family of PIAS (protein inhibitor of activated STAT) proteins. PIAS1, but not other PIAS proteins, blocked the DNA binding activity of Stat1 and inhibited Stat1-mediated gene activation in response to interferon. Coimmunoprecipitation analysis showed that PIAS1 was associated with Stat1 but not Stat2 or Stat3 after ligand stimulation. The in vivo PIAS1-Stat1 interaction requires phosphorylation of Stat1 on Tyr-701. These results identify PIAS1 as a specific inhibitor of Stat1-mediated gene activation and suggest that there may exist a specific PIAS inhibitor in every STAT signaling pathway. PMID- 9724756 TI - The Drosophila LIM-only gene, dLMO, is mutated in Beadex alleles and might represent an evolutionarily conserved function in appendage development. AB - The process of wing patterning involves precise molecular mechanisms to establish an organizing center at the dorsal-ventral boundary, which functions to direct the development of the Drosophila wing. We report that misexpression of dLMO, a Drosophila LIM-only protein, in specific patterns in the developing wing imaginal disc, disrupts the dorsal-ventral (D-V) boundary and causes errors in wing patterning. When dLMO is misexpressed along the anterior-posterior boundary, extra wing outgrowth occurs, similar to the phenotype seen when mutant clones lacking Apterous, a LIM homeodomain protein known to be essential for normal D-V patterning of the wing, are made in the wing disc. When dLMO is misexpressed along the D-V boundary in third instar larvae, loss of the wing margin is observed. This phenotype is very similar to the phenotype of Beadex, a long studied dominant mutation that we show disrupts the dLMO transcript in the 3' untranslated region. dLMO normally is expressed in the wing pouch of the third instar wing imaginal disc during patterning. A mammalian homolog of dLMO is expressed in the developing limb bud of the mouse. This indicates that LMO proteins might function in an evolutionarily conserved mechanism involved in patterning the appendages. PMID- 9724758 TI - Deletion of long-range regulatory elements upstream of SOX9 causes campomelic dysplasia. AB - Campomelic dysplasia (CD) is a rare, neonatal human chondrodysplasia characterized by bowing of the long bones and often associated with male-to female sex-reversal. Patients present with either heterozygous mutations in the SOX9 gene or chromosome rearrangements mapping at least 50 kb upstream of SOX9. Whereas mutations in SOX9 ORF cause haploinsufficiency, the effects of translocations 5' to SOX9 are unclear. To test whether these rearrangements also cause haploinsufficiency by altering spatial and temporal expression of SOX9, we generated mice transgenic for human SOX9-lacZ yeast artificial chromosomes containing variable amounts of DNA sequences upstream of SOX9. We show that elements necessary for SOX9 expression during skeletal development are highly conserved between mouse and human and reveal that a rearrangement upstream of SOX9, similar to those observed in CD patients, leads to a substantial reduction of SOX9 expression, particularly in chondrogenic tissues. These data demonstrate that important regulatory elements are scattered over a large region upstream of SOX9 and explain how particular aspects of the CD phenotype are caused by chromosomal rearrangements 5' to SOX9. PMID- 9724757 TI - The optx2 homeobox gene is expressed in early precursors of the eye and activates retina-specific genes. AB - Vertebrate eye development begins at the gastrula stage, when a region known as the eye field acquires the capacity to generate retina and lens. Optx2, a homeobox gene of the sine oculis-Six family, is selectively expressed in this early eye field and later in the lens placode and optic vesicle. The distal and ventral portion of the optic vesicle are fated to become the retina and optic nerve, whereas the dorsal portion eventually loses its neural characteristics and activates the synthesis of melanin, forming the retinal pigment epithelium. Optx2 expression is turned off in the future pigment epithelium but remains expressed in the proliferating neuroblasts and differentiating cells of the neural retina. When an Optx2-expressing plasmid is transfected into embryonic or mature chicken pigment epithelial cells, these cells adopt a neuronal morphology and express markers characteristic of developing neural retina and photoreceptors. One explanation of these results is that Optx2 functions as a determinant of retinal precursors and that it has induced the transdifferentiation of pigment epithelium into retinal neurons and photoreceptors. We also have isolated optix, a Drosophila gene that is the closest insect homologue of Optx2 and Six3. Optix is expressed during early development of the fly head and eye primordia. PMID- 9724760 TI - Top-down versus bottom-up and the Ruritanian bean bug. AB - In a recent article, Hunter uses the late George Varley and George Gradwell's long-term data on the winter moth (Operophtera brumata) and green tortrix (Tortrix viridana) populations to propose a method of quantifying the relative importance of top-down effects (because of natural enemies) and bottom-up effects (because of resource competition) in influencing population dynamics. We believe this approach is deeply flawed. Using Varley and Gradwell's winter moth study, we show that the problems with Hunter's analysis lie in his misinterpretation of the population dynamics and his inappropriate use of statistical techniques. We also emphasize the importance of distinguishing clearly between two quite different things: firstly, top-down and bottom-up regulation of populations and secondly, the much simpler task of categorizing factors affecting changes in population density as either top-down or bottom-up processes. PMID- 9724759 TI - DNase I-hypersensitive sites I and II of the human growth hormone locus control region are a major developmental activator of somatotrope gene expression. AB - High-level expression of the human growth hormone (hGH) gene is limited to somatotrope and lactosomatotrope cells of the anterior pituitary. We previously identified a locus control region (LCR) for the hGH gene composed of four tissue specific DNase I-hypersensitive sites (HS) located between -14.6 kb and -32 kb 5' to the hGH transcription start site that is responsible for establishing a physiologically regulated chromatin domain for hGH transgene expression in mouse pituitary. In the present study we demonstrated that the LCR mediates somatotrope and lactosomatotrope restriction on an otherwise weakly and diffusely expressed hGH transgene. The subregion of the LCR containing the two pituitary-specific HS, HSI and HSII (-14.6 to -16.2 kb relative to the hGH promoter and denoted HSI,II), was found to be sufficient for mediating somatotrope and lactosomatotrope restriction, for appropriately timed induction of hGH transgene expression between embryonic days 15.5 and 16.5, and for selective extinction of hGH expression in mature lactotropes. When studied by cell transfection, the HSI,II fragment selectively enhanced transcription in a presomatotrope-derived cell line, although at levels (2- to 3-fold) well below that seen in vivo. The LCR activity of the HSI,II element was therefore localized by scoring transgene expression in fetal founder pituitaries at embryonic day 18.5. The data from these studies indicated that a 404-bp segment of the HSI,II region encodes a critical subset of LCR functions, including the establishment of a productive chromatin environment, cell-specific restriction and enhancement of expression, and appropriately timed induction of the hGH transgene during embryonic development. PMID- 9724761 TI - A conserved mode of head segmentation in arthropods revealed by the expression pattern of Hox genes in a spider. AB - Chelicerates constitute a basic arthropod group with fossil representatives from as early as the Cambrian period. Embryonic development and the subdivision of the segmented body region into a prosoma and an opisthosoma are very similar in all extant chelicerates. The mode of head segmentation, however, has long been controversial. Although all other arthropod groups show a subdivision of the head region into six segments, the chelicerates are thought to have the first antennal segment missing. To examine this problem on a molecular level, we have compared the expression pattern of Hox genes in the spider Cupiennius salei with the pattern known from insects. Surprisingly, we find that the anterior expression borders of the Hox genes are in the same register and the same relative segmental position as in Drosophila. This contradicts the view that the homologue of the first antennal segment is absent in the spider. Instead, our data suggest that the cheliceral segment is homologous to the first antennal segment and the pedipalpal segment is homologous to the second antennal (or intercalary) segment in arthropods. Our finding implies that chelicerates, myriapods, crustaceans, and insects share a single mode of head segmentation, reinforcing the argument for a monophyletic origin of the arthropods. PMID- 9724762 TI - Expression of homeobox genes shows chelicerate arthropods retain their deutocerebral segment. AB - Expression patterns of six homeobox containing genes in a model chelicerate, the oribatid mite Archegozetes longisetosus, were examined to establish homology of chelicerate and insect head segments and to investigate claims that the chelicerate deutocerebral segment has been reduced or lost. engrailed (en) expression, which has been used to demonstrate the presence of segments in insects, fails to demonstrate a reduced deutocerebral segment. Expression patterns of the chelicerate homologs of the Drosophila genes Antennapedia (Antp), Sex combs reduced (Scr), Deformed (Dfd), proboscipedia (pb), and orthodenticle (otd) confirm direct correspondence of head segments. The chelicerate deutocerebral segment has not been reduced or lost. We make further inferences concerning the evolution of heads and Hox genes in arthropods. PMID- 9724763 TI - Nonsynonymous substitution in abalone sperm fertilization genes exceeds substitution in introns and mitochondrial DNA. AB - Strong positive Darwinian selection acts on two sperm fertilization proteins, lysin and 18-kDa protein, from abalone (Haliotis). To understand the phylogenetic context for this dramatic molecular evolution, we obtained sequences of mitochondrial cytochrome c oxidase subunit I (mtCOI), and genomic sequences of lysin, 18-kDa, and a G protein subunit. Based on mtDNA differentiation, four north Pacific abalone species diverged within the past 2 million years (Myr), and remaining north Pacific species diverged over a period of 4-20 Myr. Between species nonsynonymous differences in lysin and 18-kDa exons exceed nucleotide differences in introns by 3.5- to 24-fold. Remarkably, in some comparisons nonsynonymous substitutions in lysin and 18-kDa genes exceed synonymous substitutions in mtCOI. Lysin and 18-kDa intron/exon segments were sequenced from multiple red abalone individuals collected over a 1,200-km range. Only two nucleotide changes and two sites of slippage variation were detected in a total of >29,000 nucleotides surveyed. However, polymorphism in mtCOI and a G protein intron was found in this species. This finding suggests that positive selection swept one lysin allele and one 18-kDa allele to fixation. Similarities between mtCOI and lysin gene trees indicate that rapid adaptive evolution of lysin has occurred consistently through the history of the group. Comparisons with mtCOI molecular clock calibrations suggest that nonsynonymous substitutions accumulate 2-50 times faster in lysin and 18-kDa genes than in rapidly evolving mammalian genes. PMID- 9724764 TI - Proposed mechanism for stability of proteins to evolutionary mutations. AB - It is shown that the sequence-ordering tendencies induced by design into different fast-folding, thermally stable native structures interfere. This interference results in a type of quasiorthogonality between optimal native structures, which divides sequence space into fast-folding, thermally stable families surrounded by slow-folding, low stability shells. A concrete example of this effect is provided by using a simple alpha carbon type model in which a complete correspondence is established between sequence and structure. It is speculated that gaps can occur in the space of protein-like sequences separating the sequence families and resulting in a mechanism for stability and diversity of protein sequence information. PMID- 9724765 TI - Adult fitness consequences of sexual selection in Drosophila melanogaster. AB - Few experiments have demonstrated a genetic correlation between the process of sexual selection and fitness benefits in offspring, either through female choice or male competition. Those that have looked at the relationship between female choice and offspring fitness have focused on juvenile fitness components, rather than fitness at later stages in the life cycle. In addition, many of these studies have not controlled for possible maternal effects. To test for a relationship between sexual selection and adult fitness, we carried out an artificial selection experiment in the fruit fly, Drosophila melanogaster. We created two treatments that varied in the level of opportunity for sexual selection. Increased opportunity for female choice and male competition was genetically correlated with an increase in adult survivorship, as well as an increase in male and female body size. Contrary to previous, single-generation studies, we did not find an increase in larval competitive ability. This study demonstrates that mate choice and/or male-male competition are correlated with an increase in at least one adult fitness component of offspring. PMID- 9724766 TI - Multiple independent origins of mitochondrial gene order in birds. AB - Mitochondrial genomes of all vertebrate animals analyzed to date have the same 37 genes, whose arrangement in the circular DNA molecule varies only in the relative position of a few genes. This relative conservation suggests that mitochondrial gene order characters have potential utility as phylogenetic markers for higher level vertebrate taxa. We report discovery of a mitochondrial gene order that has had multiple independent originations within birds, based on sampling of 137 species representing 13 traditionally recognized orders. This provides evidence of parallel evolution in mitochondrial gene order for animals. Our results indicate operation of physical constraints on mitochondrial gene order changes and support models for gene order change based on replication error. Bird mitochondria have a displaced OL (origin of light-strand replication site) as do various other Reptilia taxa prone to gene order changes. Our findings point to the need for broad taxonomic sampling in using mitochondrial gene order for phylogenetic analyses. We found, however, that the alternative mitochondrial gene orders distinguish the two primary groups of songbirds (order Passeriformes), oscines and suboscines, in agreement with other molecular as well as morphological data sets. Thus, although mitochondrial gene order characters appear susceptible to some parallel evolution because of mechanistic constraints, they do hold promise for phylogenetic studies. PMID- 9724767 TI - Replicational and transcriptional selection on codon usage in Borrelia burgdorferi. AB - With more than 10 fully sequenced, publicly available prokaryotic genomes, it is now becoming possible to gain useful insights into genome evolution. Before the genome era, many evolutionary processes were evaluated from limited data sets and evolutionary models were constructed on the basis of small amounts of evidence. In this paper, I show that genes on the Borrelia burgdorferi genome have two separate, distinct, and significantly different codon usages, depending on whether the gene is transcribed on the leading or lagging strand of replication. Asymmetrical replication is the major source of codon usage variation. Replicational selection is responsible for the higher number of genes on the leading strands, and transcriptional selection appears to be responsible for the enrichment of highly expressed genes on these strands. Replicational transcriptional selection, therefore, has an influence on the codon usage of a gene. This is a new paradigm of codon selection in prokaryotes. PMID- 9724768 TI - Unusual horizontal transfer of a long interspersed nuclear element between distant vertebrate classes. AB - We have shown previously by Southern blot analysis that Bov-B long interspersed nuclear elements (LINEs) are present in different Viperidae snake species. To address the question as to whether Bov-B LINEs really have been transmitted horizontally between vertebrate classes, the analysis has been extended to a larger number of vertebrate, invertebrate, and plant species. In this paper, the evolutionary origin of Bov-B LINEs is shown unequivocally to be in Squamata. The previously proposed horizontal transfer of Bov-B LINEs in vertebrates has been confirmed by their discontinuous phylogenetic distribution in Squamata (Serpentes and two lizard infra-orders) as well as in Ruminantia, by the high level of nucleotide identity, and by their phylogenetic relationships. The horizontal transfer of Bov-B LINEs from Squamata to the ancestor of Ruminantia is evident from the genetic distances and discontinuous phylogenetic distribution. The ancestor of Colubroidea snakes is a possible donor of Bov-B LINEs to Ruminantia. The timing of horizontal transfer has been estimated from the distribution of Bov B LINEs in Ruminantia and the fossil data of Ruminantia to be 40-50 My ago. The phylogenetic relationships of Bov-B LINEs from the various Squamata species agrees with that of the species phylogeny, suggesting that Bov-B LINEs have been maintained stably by vertical transmission since the origin of Squamata in the Mesozoic era. PMID- 9724769 TI - A general procedure for locating and analyzing protein-binding sequence motifs in nucleic acids. AB - In the last decade, two tools, one drawn from information theory and the other from artificial neural networks, have proven particularly useful in many different areas of sequence analysis. The work presented herein indicates that these two approaches can be joined in a general fashion to produce a very powerful search engine that is capable of locating members of a given nucleic acid sequence family in either local or global sequence searches. This program can, in turn, be queried for its definition of the motif under investigation, ranking each base in context for its contribution to membership in the motif family. In principle, the method used can be applied to any binding motif, including both DNA and RNA sequence families, given sufficient family size. PMID- 9724770 TI - Tn5/IS50 target recognition. AB - This communication reports an analysis of Tn5/IS50 target site selection by using an extensive collection of Tn5 and IS50 insertions in two relatively small regions of DNA (less than 1 kb each). For both regions data were collected resulting from in vitro and in vivo transposition events. Since the data sets are consistent and transposase was the only protein present in vitro, this demonstrates that target selection is a property of only transposase. There appear to be two factors governing target selection. A target consensus sequence, which presumably reflects the target selection of individual pairs of Tn5/IS50 bound transposase protomers, was deduced by analyzing all insertion sites. The consensus Tn5/IS50 target site is A-GNTYWRANC-T. However, we observed that independent insertion sites tend to form groups of closely located insertions (clusters), and insertions very often were spaced in a 5-bp periodic fashion. This suggests that Tn5/IS50 target selection is facilitated by more than two transposase protomers binding to the DNA, and, thus, for a site to be a good target, the overlapping neighboring DNA should be a good target, too. Synthetic target sequences were designed and used to test and confirm this model. PMID- 9724771 TI - The APC variants I1307K and E1317Q are associated with colorectal tumors, but not always with a family history. AB - Classical familial adenomatous polyposis (FAP) is a high-penetrance autosomal dominant disease that predisposes to hundreds or thousands of colorectal adenomas and carcinoma and that results from truncating mutations in the APC gene. A variant of FAP is attenuated adenomatous polyposis coli, which results from germ line mutations in the 5' and 3' regions of the APC gene. Attenuated adenomatous polyposis coli patients have "multiple" colorectal adenomas (typically fewer than 100) without the florid phenotype of classical FAP. Another group of patients with multiple adenomas has no mutations in the APC gene, and their phenotype probably results from variation at a locus, or loci, elsewhere in the genome. Recently, however, a missense variant of APC (I1307K) was described that confers an increased risk of colorectal tumors, including multiple adenomas, in Ashkenazim. We have studied a set of 164 patients with multiple colorectal adenomas and/or carcinoma and analyzed codons 1263-1377 (exon 15G) of the APC gene for germ-line variants. Three patients with the I1307K allele were detected, each of Ashkenazi descent. Four patients had a germ-line E1317Q missense variant of APC that was not present in controls; one of these individuals had an unusually large number of metaplastic polyps of the colorectum. There is increasing evidence that there exist germ-line variants of the APC gene that predispose to the development of multiple colorectal adenomas and carcinoma, but without the florid phenotype of classical FAP, and possibly with importance for colorectal cancer risk in the general population. PMID- 9724772 TI - An SmtB-like repressor from Synechocystis PCC 6803 regulates a zinc exporter. AB - ORF slr0798, now designated ziaA, from Synechocystis PCC 6803 encodes a polypeptide with sequence features of heavy metal transporting P-type ATPases. Increased Zn2+ tolerance and reduced 65Zn accumulation was observed in Synechococcus PCC 7942, strain R2-PIM8(smt), containing ziaA and upstream regulatory sequences, compared with control cells. Conversely, reduced Zn2+ tolerance was observed following disruption of ziaA in Synechocystis PCC 6803, and ziaA-mediated restoration of Zn2+ tolerance has subsequently been used as a selectable marker for transformation. Nucleotide sequences upstream of ziaA, fused to a promoterless lacZ gene, conferred Zn2+-dependent beta-galactosidase activity when introduced into R2-PIM8(smt). The product of ORF sll0792, designated ZiaR, is a Zn2+-responsive repressor of ziaA transcription. Reporter gene constructs lacking ziaR conferred elevated Zn2+-independent expression from the ziaA operator-promoter in R2-PIM8(smt). Gel retardation assays detected ZiaR dependent complexes forming with the zia operator-promoter and ZiaR-DNA binding was enhanced by treatment with a metal-chelator in vitro. Two mutants of ZiaR (C71S/C73S and H116R) bound to, and repressed expression from, the ziaA operator promoter but were unable to sense Zn2+. Metal coordination to His-imidazole and Cys-thiolate ligands at these residues of ZiaR is thus implicated in Zn2+ perception by Synechocystis PCC 6803. PMID- 9724773 TI - Increased anxiety of mice lacking the serotonin1A receptor. AB - Brain serotonin (5-HT) has been implicated in a number of physiological processes and pathological conditions. These effects are mediated by at least 14 different 5-HT receptors. We have inactivated the gene encoding the 5-HT1A receptor in mice and found that receptor-deficient animals have an increased tendency to avoid a novel and fearful environment and to escape a stressful situation, behaviors consistent with an increased anxiety and stress response. Based on the role of the 5-HT1A receptor in the feedback regulation of the 5-HT system, we hypothesize that an increased serotonergic neurotransmission is responsible for the anxiety like behavior of receptor-deficient animals. This view is consistent with earlier studies showing that pharmacological activation of the 5-HT system is anxiogenic in animal models and also in humans. PMID- 9724774 TI - Two modes of transvection: enhancer action in trans and bypass of a chromatin insulator in cis. AB - Ed Lewis introduced the term "transvection" in 1954 to describe mechanisms that can cause the expression of a gene to be sensitive to the proximity of its homologue. Transvection since has been reported at an increasing number of loci in Drosophila, where homologous chromosomes are paired in somatic tissues, as well as at loci in other organisms. At the Drosophila yellow gene, transvection can explain intragenic complementation involving the yellow2 allele (y2). Here, transvection was proposed to occur by enhancers of one allele acting in trans on the promoter of a paired homologue. In this report, we describe two yellow alleles that strengthen this model and reveal an unexpected, second mechanism for transvection. Data suggest that, in addition to enhancer action in trans, transvection can occur by enhancer bypass of a chromatin insulator in cis. We propose that bypass results from the topology of paired genes. Finally, transvection at yellow can occur in genotypes not involving y2, implying that it is a feature of yellow itself and not an attribute of one particular allele. PMID- 9724775 TI - New susceptibility locus for rheumatoid arthritis suggested by a genome-wide linkage study. AB - Rheumatoid arthritis (RA), the most common autoimmune disease, is associated in families with other autoimmune diseases, including insulin-dependent diabetes mellitus (IDDM). Its genetic component has been suggested by familial aggregation (lambdas = 5), twin studies, and segregation analysis. HLA, which is the only susceptibility locus known, has been estimated to account for one-third of this component. The aim of this paper was to identify new RA loci. A genome scan was performed with 114 European Caucasian RA sib pairs from 97 nuclear families. Linkage was significant only for HLA (P < 2.5.10(-5)) and nominal for 19 markers in 14 other regions (P < 0.05). Four of the loci implicated in IDDM potentially overlap with these regions: the putative IDDM6, IDDM9, IDDM13, and DXS998 loci. The first two of these candidate regions, defined in the RA genome scan by the markers D18S68-D18S61-D18S469 (18q22-23) and D3S1267 (3q13), respectively, were studied in 194 additional RA sib pairs from 164 nuclear families. Support for linkage to chromosome 3 only was extended significantly (P = 0.002). The analysis of all 261 families provided a linkage evidence of P = 0. 001 and suggested an interaction between this putative RA locus and HLA. This locus could account for 16% of the genetic component of RA. Candidate genes include those coding for CD80 and CD86, molecules involved in antigen-specific T cell recognition. In conclusion, this first genome scan in RA Caucasian families revealed 14 candidate regions, one of which was supported further by the study of a second set of families. PMID- 9724776 TI - The reductive enzyme thioredoxin 1 acts as an oxidant when it is exported to the Escherichia coli periplasm. AB - Thioredoxin 1 is a major thiol-disulfide oxidoreductase in the cytoplasm of Escherichia coli. One of its functions is presumed to be the reduction of the disulfide bond in the active site of the essential enzyme ribonucleotide reductase. Thioredoxin 1 is kept in a reduced state by thioredoxin reductase. In a thioredoxin reductase null mutant however, most of thioredoxin 1 is in the oxidized form; recent reports have suggested that this oxidized form might promote disulfide bond formation in vivo. In the Escherichia coli periplasm, the protein disulfide isomerase DsbC is maintained in the reduced and active state by the membrane protein DsbD. In a dsbD null mutant, DsbC accumulates in the oxidized form. This oxidized form is then able to promote disulfide bond formation. In both these cases, the inversion of the function of these thiol oxidoreductases appears to be due to an altered redox balance of the environment in which they find themselves. Here, we show that thioredoxin 1 attached to the alkaline phosphatase signal sequence can be exported into the E. coli periplasm. In this new environment for thioredoxin 1, we show that thioredoxin 1 can promote disulfide bond formation and, therefore, partially complement a dsbA strain defective for disulfide bond formation. Thus, we provide evidence that by changing the location of thioredoxin 1 from cytoplasm to periplasm, we change its function from a reductant to an oxidant. We conclude that the in vivo redox function of thioredoxin 1 depends on the redox environment in which it is localized. PMID- 9724777 TI - PAGE-1, an X chromosome-linked GAGE-like gene that is expressed in normal and neoplastic prostate, testis, and uterus. AB - We have used a combination of computerized database mining and experimental expression analyses to identify a gene that is preferentially expressed in normal male and female reproductive tissues, prostate, testis, fallopian tube, uterus, and placenta, as well as in prostate cancer, testicular cancer, and uterine cancer. This gene is located on the human X chromosome, and it is homologous to a family of genes encoding GAGE-like proteins. GAGE proteins are expressed in a variety of tumors and in testis. We designate the novel gene PAGE-1 because the expression pattern in the Cancer Genome Anatomy Project libraries indicates that it is predominantly expressed in normal and neoplastic prostate. Further database analysis indicates the presence of other genes with high homology to PAGE-1, which were found in cDNA libraries derived from testis, pooled libraries (with testis), and in a germ cell tumor library. The expression of PAGE-1 in normal and malignant prostate, testicular, and uterine tissues makes it a possible target for the diagnosis and possibly for the vaccine-based therapy of neoplasms of prostate, testis, and uterus. PMID- 9724778 TI - A cis-acting element that directs the activity of the murine methylation modifier locus Ssm1. AB - Silencing of chromosomal domains has been described in diverse systems such as position effect variegation in insects, silencing near yeast telomeres, and mammalian X chromosome inactivation. In mammals, silencing is associated with methylation at CpG dinucleotides, but little is known about how methylation patterns are established or altered during development. We previously described a strain-specific modifier locus, Ssm1, that controls the methylation of a complex transgene. In this study we address the questions of the nature of Ssm1's targets and whether its effect extends into adjacent sequences. By examining the inheritance of methylation patterns in a series of mice harboring deletion derivatives of the original transgene, we have identified a discrete segment, derived from the gpt gene of Escherichia coli, that is a major determinant for Ssm1-mediated methylation. Methylation analysis of sequences adjacent to a transgenic target indicates that the influence of this modifier extends into the surrounding chromosome in a strain-dependent fashion. Implications for the mechanism of Ssm1 action are discussed. PMID- 9724779 TI - Chromosomal transposition of a Tc1/mariner-like element in mouse embryonic stem cells. AB - Mouse has become an increasingly important organism for modeling human diseases and for determining gene function in a mammalian context. Unfortunately, transposon-tagged mutagenesis, one of the most valuable tools for functional genomics, still is not available in this organism. On the other hand, it has long been speculated that members of the Tc1/mariner-like elements may be less dependent on host factors and, hence, can be introduced into heterologous organisms. However, this prediction has not been realized in mice. We report here the chromosomal transposition of the Sleeping Beauty (SB) element in mouse embryonic stem cells, providing evidence that it can be used as an in vivo mutagen in mice. PMID- 9724780 TI - Equilibrium distributions of microsatellite repeat length resulting from a balance between slippage events and point mutations. AB - We describe and test a Markov chain model of microsatellite evolution that can explain the different distributions of microsatellite lengths across different organisms and repeat motifs. Two key features of this model are the dependence of mutation rates on microsatellite length and a mutation process that includes both strand slippage and point mutation events. We compute the stationary distribution of allele lengths under this model and use it to fit DNA data for di-, tri-, and tetranucleotide repeats in humans, mice, fruit flies, and yeast. The best fit results lead to slippage rate estimates that are highest in mice, followed by humans, then yeast, and then fruit flies. Within each organism, the estimates are highest in di-, then tri-, and then tetranucleotide repeats. Our estimates are consistent with experimentally determined mutation rates from other studies. The results suggest that the different length distributions among organisms and repeat motifs can be explained by a simple difference in slippage rates and that selective constraints on length need not be imposed. PMID- 9724782 TI - A knob-associated tandem repeat in maize capable of forming fold-back DNA segments: are chromosome knobs megatransposons? AB - A class of tandemly repeated DNA sequences (TR-1) of 350-bp unit length was isolated from the knob DNA of chromosome 9 of Zea mays L. Comparative fluorescence in situ hybridization revealed that TR-1 elements are also present in cytologically detectable knobs on other maize chromosomes in different proportions relative to the previously described 180-bp repeats. At least one knob on chromosome 4 is composed predominantly of the TR-1 repeat. In addition, several small clusters of the TR-1 and 180-bp repeats have been found in different chromosomes, some not located in obvious knob heterochromatin. Variation in restriction fragment fingerprints and copy number of the TR-1 elements was found among maize lines and among maize chromosomes. TR-1 tandem arrays up to 70 kilobases in length can be interspersed with stretches of 180-bp tandem repeat arrays. DNA sequence analysis and restriction mapping of one particular stretch of tandemly arranged TR-1 units indicate that these elements may be organized in the form of fold-back DNA segments. The TR-1 repeat shares two short segments of homology with the 180-bp repeat. The longest of these segments (31 bp; 64% identity) corresponds to the conserved region among 180-bp repeats. The polymorphism and complex structure of knob DNA suggest that, similar to the fold-back DNA-containing giant transposons in Drosophila, maize knob DNA may have some properties of transposable elements. PMID- 9724781 TI - A modifier screen in the eye reveals control genes for Kruppel activity in the Drosophila embryo. AB - Irregular facets (If) is a dominant mutation of Drosophila that results in small eyes with fused ommatidia. Previous results showed that the gene Kruppel (Kr), which is best known for its early segmentation function, is expressed ectopically in If mutant eye discs. However, it was not known whether ectopic Kr activity is either the cause or the result of the If mutation. Here, we show that If is a gain-of-function allele of Kr. We then used the If mutation in a genetic screen to identify dominant enhancers and suppressors of Kr activity on the third chromosome. Of 30 identified Kr-interacting loci, two were cloned, and we examined whether they also represent components of a natural Kr-dependent developmental pathway of the embryo. We show that the two genes, eyelid (eld) and extramacrochaetae (emc), which encode a Bright family-type DNA binding protein and a helix-loop-helix factor, respectively, are necessary to achieve the singling-out of a unique Kr-expressing cell during the development of the Malpighian tubules, the excretory organs of the fly. The results indicate that the Kr gain-of-function mutation If provides a tool to identify genes that are active during eye development and that a number of them function also in the control of Kr-dependent developmental processes. PMID- 9724783 TI - Determination of gene organization in individual haplotypes by analyzing single DNA fragments from single spermatozoa. AB - To determine human Ig heavy chain variable region (VH) gene segment organization on individual homologous chromosomes, an efficient approach has been developed. Single spermatozoa were used as subjects for the study. Upon sperm lysis, VH regions in each sperm were randomly sheared into fragments by the random Brownian force. The fragments were separated from each other by aliquoting the lysate into a certain number of tubes. The gene segments in the VH1 and VH4 families in each tube were identified by denaturing gradient gel electrophoresis after PCR amplification. The polymorphic VH sequences were used to determine the parental origins of the analyzed sperm. VH segment organization in the parental haplotypes was determined by aligning the overlapping fragments from the spermatozoa with the corresponding haplotypes. Based on this comparison between the resulting haplotype maps and the composite map reported previously, the VH region on chromosome 14 could be subdivided into four portions. The numbers and compositions of the VH gene segments differ considerably among the maps in two portions, but are highly conserved in the other two. The data also indicate that the VH region on chromosome 15 may contain a large duplicated block with copy number varying among haplotypes. The approach used in the present study may be used to construct high-resolution haplotype maps without molecular cloning. PMID- 9724784 TI - Identification of HLA class II-restricted determinants of Mycobacterium tuberculosis-derived proteins by using HLA-transgenic, class II-deficient mice. AB - T helper 1 cells play a major role in protective immunity against mycobacterial pathogens. Since the antigen (Ag) specificity of CD4(+) human T cells is strongly controlled by HLA class II polymorphism, the immunogenic potential of candidate Ags needs to be defined in the context of HLA polymorphism. We have taken advantage of class II-deficient (Ab0) mice, transgenic for either HLA-DRA/B1*0301 (DR3) or HLA-DQB1*0302/DQA*0301 (DQ8) alleles. In these animals, all CD4(+) T cells are restricted by the HLA molecule. We reported previously that human DR3 restricted T cells frequently recognize heat shock protein (hsp)65 of Mycobacterium tuberculosis, and only a single hsp65 epitope, p1-20. DR3.Ab0 mice, immunized with bacillus Calmette-Guerin or hsp65, developed T cell responses to M. tuberculosis, and recognized the same hsp65 epitope, p1-20. Hsp65-immunized DQ8.Ab0 mice mounted a strong response to bacillus Calmette-Guerin but not to p1 20. Instead, we identified three new DQ8-restricted T cell epitopes in the regions 171-200, 311-340, and 411-440. DR3.Ab0 mice immunized with a second major M. tuberculosis protein, Ag85 (composed of 85A, 85B, and 85C), also developed T cell responses against only one determinant, 85B p51-70, that was identified in this study. Importantly, subsequent analysis of human T cell responses revealed that HLA-DR3+, Ag85-reactive individuals recognize exactly the same peptide epitope as DR3.Ab0 mice. Strikingly, both DR3-restricted T cell epitopes represent the best DR3-binding sequences in hsp65 and 85B, revealing a strong association between peptide-immunodominance and HLA binding affinity. Immunization of DR3.Ab0 with the immunodominant peptides p1-20 and p51-70 induced T cell reactivity to M. tuberculosis. Thus, for two different Ags, T cells from DR3.Ab0 mice and HLA-DR3+ humans recognize the same immunodominant determinants. Our data support the use of HLA-transgenic mice in identifying human T cell determinants for the design of new vaccines. PMID- 9724785 TI - beta-chemokines and neutralizing antibody titers correlate with sterilizing immunity generated in HIV-1 vaccinated macaques. AB - One of the obstacles to AIDS vaccine development is the variability of HIV-1 within individuals and within infected populations, enabling viral escape from highly specific vaccine induced immune responses. An understanding of the different immune mechanisms capable of inhibiting HIV infection may be of benefit in the eventual design of vaccines effective against HIV-1 variants. To study this we first compared the immune responses induced in Rhesus monkeys by using two different immunization strategies based on the same vaccine strain of HIV-1. We then utilized a chimeric simian/HIV that expressed the envelope of a dual tropic HIV-1 escape variant isolated from a later time point from the same patient from which the vaccine strain was isolated. Upon challenge, one vaccine group was completely protected from infection, whereas all of the other vaccinees and controls became infected. Protected macaques developed highest titers of heterologous neutralizing antibodies, and consistently elevated HIV-1-specific T helper responses. Furthermore, only protected animals had markedly increased concentrations of RANTES, macrophage inflammatory proteins 1alpha and 1beta produced by circulating CD8(+) T cells. These results suggest that vaccine strategies that induce multiple effector mechanisms in concert with beta chemokines may be desired in the generation of protective immune responses by HIV 1 vaccines. PMID- 9724786 TI - Antigenicity of fullerenes: antibodies specific for fullerenes and their characteristics. AB - The recent interest in using Buckminsterfullerene (fullerene) derivatives in biological systems raises the possibility of their assay by immunological procedures. This, in turn, leads to the question of the ability of these unprecedented polygonal structures, made up solely of carbon atoms, to induce the production of specific antibodies. Immunization of mice with a C60 fullerene derivative conjugated to bovine thyroglobulin yielded a population of fullerene specific antibodies of the IgG isotype, showing that the immune repertoire was diverse enough to recognize and process fullerenes as protein conjugates. The population of antibodies included a subpopulation that crossreacted with a C70 fullerene as determined by immune precipitation and ELISA procedures. These assays were made possible by the synthesis of water-soluble fullerene derivatives, including bovine and rabbit serum albumin conjugates and derivatives of trilysine and pentalysine, all of which were characterized as to the extent of substitution and their UV-Vis spectra. Possible interactions of fullerenes with the combining sites of IgG are discussed based on the physical chemistry of fullerenes and previously described protein-fullerene interactions. They remain to be confirmed by the isolation of mAbs for x-ray crystallographic studies. PMID- 9724787 TI - Generation of mucosal cytotoxic T cells against soluble protein by tissue specific environmental and costimulatory signals. AB - We compared peripheral and mucosal primary CD8 T cell responses to inflammatory and noninflammatory forms of antigen in a T cell-adoptive transfer system. Immunization with the soluble antigen, ovalbumin (ova), administered i.p. or orally without adjuvant, activated nonmucosal CD8 T cells but did not induce cytotoxic activity. However, after activation, the transferred cells entered the intestinal mucosa and became potent antigen-specific killers. Thus, exogenous intact soluble protein entered the major histocompatibility complex class I antigen presentation pathway and induced mucosal cytotoxic T lymphocytes. Moreover, distinct costimulatory requirements for activation of peripheral versus mucosal T cells were noted in that the CD28 ligand, B7-1, was critical for activated mucosal T cell generation but not for activation of peripheral CD8 T cells. The costimulator, B7-2, was required for optimum activation of both populations. Infection with a new recombinant vesicular stomatitis virus encoding ovalbumin induced lytic activity in mucosal as well as peripheral sites, demonstrating an adjuvant effect of inflammatory mediators produced during virus infection. Generation of antiviral cytotoxic T lymphocytes was also costimulation dependent. The results indicated that induction of peripheral tolerance via antigen administration may not extend to mucosal sites because of distinct costimulatory and inflammatory signals in the mucosa. PMID- 9724788 TI - Endothelial cell death, angiogenesis, and microvascular function after castration in an androgen-dependent tumor: role of vascular endothelial growth factor. AB - The sequence of events that leads to tumor vessel regression and the functional characteristics of these vessels during hormone-ablation therapy are not known. This is because of the lack of an appropriate animal model and monitoring technology. By using in vivo microscopy and in situ molecular analysis of the androgen-dependent Shionogi carcinoma grown in severe combined immunodeficient mice, we show that castration of these mice leads to tumor regression and a concomitant decrease in vascular endothelial growth factor (VEGF) expression. Androgen withdrawal is known to induce apoptosis in Shionogi tumor cells. Surprisingly, tumor endothelial cells begin to undergo apoptosis before neoplastic cells, and rarefaction of tumor vessels precedes the decrease in tumor size. The regressing vessels begin to exhibit normal phenotype, i.e., lower diameter, tortuosity, vascular permeability, and leukocyte adhesion. Two weeks after castration, a second wave of angiogenesis and tumor growth begins with a concomitant increase in VEGF expression. Because human tumors often relapse following hormone-ablation therapy, our data suggest that these patients may benefit from combined anti-VEGF therapy. PMID- 9724789 TI - A resistant genetic background leading to incomplete penetrance of intestinal neoplasia and reduced loss of heterozygosity in ApcMin/+ mice. AB - Previous studies of Min/+ (multiple intestinal neoplasia) mice on a sensitive genetic background, C57BL/6 (B6), showed that adenomas have lost heterozygosity for the germ-line ApcMin mutation in the Apc (adenomatous polyposis coli) gene. We now report that on a strongly resistant genetic background, AKR/J (AKR), Min induced adenoma multiplicity is reduced by about two orders of magnitude compared with that observed on the B6 background. Somatic treatment with a strong mutagen increases tumor number in AKR Min/+ mice in an age-dependent manner, similar to results previously reported for B6 Min/+ mice. Immunohistochemical analyses indicate that Apc expression is suppressed in all intestinal tumors from both untreated and treated AKR Min/+ mice. However, the mechanism of Apc inactivation in AKR Min/+ mice often differs from that observed for B6 Min/+ mice. Although loss of heterozygosity is observed in some tumors, a significant percentage of tumors showed neither loss of heterozygosity nor Apc truncation mutations. These results extend our understanding of the effects of genetic background on Min induced tumorigenesis in several ways. First, the AKR strain carries modifiers of Min in addition to Mom1. This combination of AKR modifiers can almost completely suppress spontaneous intestinal tumorigenesis associated with the Min mutation. Second, even on such a highly resistant genetic background, tumor formation continues to involve an absence of Apc function. The means by which Apc function is inactivated is affected by genetic background. Possible scenarios are discussed. PMID- 9724790 TI - In vivo magnetic resonance vascular imaging using laser-polarized 3He microbubbles. AB - Laser-polarized gases (3He and 129Xe) are currently being used in magnetic resonance imaging as strong signal sources that can be safely introduced into the lung. Recently, researchers have been investigating other tissues using 129Xe. These studies use xenon dissolved in a carrier such as lipid vesicles or blood. Since helium is much less soluble than xenon in these materials, 3He has been used exclusively for imaging air spaces. However, considering that the signal of 3He is more than 10 times greater than that of 129Xe for presently attainable polarization levels, this work has focused on generating a method to introduce 3He into the vascular system. We addressed the low solubility issue by producing suspensions of 3He microbubbles. Here, we provide the first vascular images obtained with laser-polarized 3He. The potential increase in signal and absence of background should allow this technique to produce high-resolution angiographic images. In addition, quantitative measurements of blood flow velocity and tissue perfusion will be feasible. PMID- 9724791 TI - Synthesis and biological activities of potent peptidomimetics selective for somatostatin receptor subtype 2. AB - A series of nonpeptide somatostatin agonists which bind selectively and with high affinity to somatostatin receptor subtype 2 (sst2) have been synthesized. One of these compounds, L-054,522, binds to human sst2 with an apparent dissociation constant of 0.01 nM and at least 3,000-fold selectivity when evaluated against the other somatostatin receptors. L-054,522 is a full agonist based on its inhibition of forskolin-stimulated adenylate cyclase activity in Chinese hamster ovary-K1 cells stably expressing sst2. L-054,522 has a potent inhibitory effect on growth hormone release from rat primary pituitary cells and glucagon release from isolated mouse pancreatic islets. Intravenous infusion of L-054,522 to rats at 50 microgram/kg per hr causes a rapid and sustained reduction in growth hormone to basal levels. The high potency and selectivity of L-054, 522 for sst2 will make it a useful tool to further characterize the physiological functions of this receptor subtype. PMID- 9724792 TI - Conversion of a radioresistant phenotype to a more sensitive one by disabling erbB receptor signaling in human cancer cells. AB - Inhibition of cell growth and transformation can be achieved in transformed glial cells by disabling erbB receptor signaling. However, recent evidence indicates that the induction of apoptosis may underlie successful therapy of human cancers. In these studies, we examined whether disabling oncoproteins of the erbB receptor family would sensitize transformed human glial cells to the induction of genomic damage by gamma-irradiation. Radioresistant human glioblastoma cells in which erbB receptor signaling was inhibited exhibited increased growth arrest and apoptosis in response to DNA damage. Apoptosis was observed after radiation in human glioma cells containing either a wild-type or mutated p53 gene product and suggested that both p53-dependent and -independent mechanisms may be responsible for the more radiosensitive phenotype. Because cells exhibiting increased radiation-induced apoptosis were also capable of growth arrest in serum-deprived conditions and in response to DNA damage, apoptotic cell death was not induced simply as a result of impaired growth arrest pathways. Notably, inhibition of erbB signaling was a more potent stimulus for the induction of apoptosis than prolonged serum deprivation. Proximal receptor interactions between erbB receptor members thus influence cell cycle checkpoint pathways activated in response to DNA damage. Disabling erbB receptors may improve the response to gamma irradiation and other cytotoxic therapies, and this approach suggests that present anticancer strategies could be optimized. PMID- 9724793 TI - Identification of inflections in T-cell counts among HIV-1-infected individuals and relationship with progression to clinical AIDS. AB - Studies of circulating T (CD3(+)) lymphocytes have shown that on a population basis T-cell numbers remain stable for many years after HIV-1 infection (blind T cell homeostasis), but decline rapidly beginning approximately 1.5-2.5 years before the onset of clinical AIDS. We derived a general method for defining the loss of homeostasis on the individual level and for determining the prevalence of homeostasis loss according to HIV status and the occurrence of AIDS in more than 5,000 men enrolled in the Multicenter AIDS Cohort Study. We used a segmented regression model for log10 CD3(+) cell counts that included separate T-cell trajectories before and after a time (the T-cell inflection point) where the loss of T-cell homeostasis was most likely to have occurred. The average slope of CD3(+) lymphocyte counts before the inflection point was close to zero for HIV- and HIV+ men, consistent with blind T-cell homeostasis. After the inflection point, the HIV+ individuals who developed AIDS generally showed a dramatic decline in CD3(+) cell counts relative to HIV- men and HIV+ men not developing AIDS. A CD3(+) cell decline of greater than 10 percent per year was present in 77% of HIV+ men developing AIDS but in only 23% of HIV+ men with no onset of AIDS. Our findings at the individual level support the blind T-cell homeostasis hypothesis and provide strong evidence that the loss of homeostasis is an important mechanism in the pathogenesis of the severe immunodeficiency that characterizes the late stages of HIV infection. PMID- 9724794 TI - Functional expression of the Wilson disease protein reveals mislocalization and impaired copper-dependent trafficking of the common H1069Q mutation. AB - Wilson disease is an autosomal recessive disorder of hepatic copper metabolism caused by mutations in a gene encoding a copper-transporting P-type ATPase. To elucidate the function of the Wilson protein, wild-type and mutant Wilson cDNAs were expressed in a Menkes copper transporter-deficient mottled fibroblast cell line defective in copper export. Expression of the wild-type cDNA demonstrated trans-Golgi network localization and copper-dependent trafficking of the Wilson protein identical to previous observations for the endogenously expressed protein in hepatocytes. Furthermore, expression of the Wilson cDNA rescued the mottled phenotype as evidenced by a reduction in copper accumulation and restoration of cell viability. In contrast, expression of an H1069Q mutant Wilson cDNA did not rescue the mottled phenotype, and immunofluorescence studies showed that this mutant Wilson protein was localized in the endoplasmic reticulum. Consistent with these findings, pulse-chase analysis demonstrated a 5-fold decrease in the half life of the H1069Q mutant as compared with the wild-type protein. Maintenance of these transfected cell lines at 28 degreesC resulted in localization of the H1069Q protein in the trans-Golgi network, suggesting that a temperature sensitive defect in protein folding followed by degradation constitutes the molecular basis of Wilson disease in patients harboring the H1069Q mutation. Taken together, these studies describe a tractable expression system for elucidating the function and localization of the copper-transporting ATPases in mammalian cells and provide compelling evidence that the Wilson protein can functionally substitute for the Menkes protein, supporting the concept that these proteins use common biochemical mechanisms to effect cellular copper homeostasis. PMID- 9724795 TI - ETO, fusion partner in t(8;21) acute myeloid leukemia, represses transcription by interaction with the human N-CoR/mSin3/HDAC1 complex. AB - The t(8;21) translocation between two genes known as AML1 and ETO is seen in approximately 12-15% of all acute myeloid leukemia (AML) and is the second-most frequently observed nonrandom genetic alteration associated with AML. AML1 up regulates a number of target genes critical to normal hematopoiesis, whereas the AML1/ETO fusion interferes with this trans-activation. We discovered that the fusion partner ETO binds to the human homolog of the murine nuclear receptor corepressor (N-CoR). The interaction is mediated by two unusual zinc finger motifs present at the carboxyl terminus of ETO. Human N-CoR (HuN-CoR), which we cloned and sequenced in its entirety, encodes a 2,440-amino acid polypeptide and has a central domain that binds ETO. N-CoR, mammalian Sin3 (mSin3A and B), and histone deacetylase 1 (HDAC1) form a complex that alters chromatin structure and mediates transcriptional repression by nuclear receptors and by a number of oncoregulatory proteins. We found that ETO, through its interaction with the N CoR/mSin3/HDAC1 complex, is also a potent repressor of transcription. This observation provides a mechanism for how the AML1/ETO fusion may inhibit expression of AML1-responsive target genes and disturb normal hematopoiesis. PMID- 9724796 TI - A viral gene that activates lytic cycle expression of Kaposi's sarcoma-associated herpesvirus. AB - Herpesviruses exist in two states, latency and a lytic productive cycle. Here we identify an immediate-early gene encoded by Kaposi's sarcoma-associated herpesvirus (KSHV)/human herpesvirus eight (HHV8) that activates lytic cycle gene expression from the latent viral genome. The gene is a homologue of Rta, a transcriptional activator encoded by Epstein-Barr virus (EBV). KSHV/Rta activated KSHV early lytic genes, including virus-encoded interleukin 6 and polyadenylated nuclear RNA, and a late gene, small viral capsid antigen. In cells dually infected with Epstein-Barr virus and KSHV, each Rta activated only autologous lytic cycle genes. Expression of viral cytokines under control of the KSHV/Rta gene is likely to contribute to the pathogenesis of KSHV-associated diseases. PMID- 9724797 TI - Identification of the gene (lgtG) encoding the lipooligosaccharide beta chain synthesizing glucosyl transferase from Neisseria gonorrhoeae. AB - The lipooligosaccharide from Neisseria gonorrhoeae (GC), consists of lipid A, an oligosaccharide core and three branches, alpha, beta, and gamma. We report the cloning of the gene (lgtG, lipooligosaccharide glycosyl transferase G) encoding the glucosyl transferase of GC that initiates the beta chain which consists of a lactosyl moiety. This gene contains a homopolymeric tract of cytidine [poly(C)] and we demonstrate that changes in the number of Cs in poly(C) account for the variation of beta chain expression in different GC strains. Biochemical analyses and mass spectrometry clearly attribute the reactivity of mAb 2C7 to the presence of the lactosyl beta chain. In addition, we demonstrate that in the absence of the lactosyl group, a phosphoethanolamine is added to generate a new antigenic epitope as evidenced by the gain of reactivity to mAb 2-L1-8. These results show that, like the alpha chain, the beta chain of lipooligosaccharide is subject to antigenic variation. PMID- 9724798 TI - Segment-specific pattern of sympathetic preganglionic projections in the chicken embryo spinal cord is altered by retinoids. AB - Sympathetic preganglionic neurons exhibit segment-specific projections. Preganglionic neurons located in rostral spinal segments project rostrally within the sympathetic chain, those located in caudal spinal segments project caudally, and those in midthoracic segments project either rostrally or caudally in segmentally graded proportions. Moreover, rostrally and caudally projecting preganglionic neurons are skewed toward the rostral and caudal regions, respectively, of each midthoracic segment. The mechanisms that establish these segment-specific projections are unknown. Here we show that experimental manipulation of retinoid signaling in the chicken embryo alters the segment specific pattern of sympathetic preganglionic projections and that this effect is mediated by the somitic mesoderm. Application of exogenous retinoic acid to a single rostral thoracic somite decreases the number of rostrally projecting preganglionic neurons at that level. Conversely, disrupting endogenous synthesis of retinoic acid in a single caudal thoracic somite increases the number of rostrally projecting preganglionic neurons at that level. The number of caudally projecting neurons does not change in either case, indicating that the effect is specific for rostrally projecting preganglionic neurons. These results indicate that the sizes of the rostrally and caudally projecting populations may be independently regulated by different factors. Opposing gradients of such factors along the longitudinal axis of the thoracic region of the embryo could be sufficient, in combination, to determine the segment-specific identity of preganglionic projections. PMID- 9724799 TI - Expression of a dominant negative TrkB receptor, T1, reveals a requirement for presynaptic signaling in BDNF-induced synaptic potentiation in cultured hippocampal neurons. AB - We have developed a method to analyze the relative contributions of pre- and postsynaptic actions of a particular gene product in neurons in culture and potentially in slices using adenovirus-mediated gene transfer. A recombinant virus directed the expression of both a GFP reporter protein and TrkB.T1, a C terminal truncated dominant negative TrkB neurotrophin receptor. When expressed in the presynaptic cell at synapses between embryonic hippocampal neurons in culture, the dominant negative TrkB.T1 inhibited two forms of synaptic potentiation induced by the neurotrophin brain-derived neurotrophic factor (BDNF): (i) greater evoked synaptic transmission and (ii) higher frequency of spontaneous miniature synaptic currents. These inhibition effects are not seen if the transgene is expressed only in the postsynaptic cell. We conclude that BDNF TrkB signal transduction in the presynaptic terminal leads to both types of potentiation and is therefore the primary cause of synaptic enhancement by BDNF in these neurons. PMID- 9724800 TI - Characterization of a calmodulin kinase II inhibitor protein in brain. AB - Ca2+/calmodulin-dependent protein kinase II (CaM-KII) regulates numerous physiological functions, including neuronal synaptic plasticity through the phosphorylation of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid-type glutamate receptors. To identify proteins that may interact with and modulate CaM KII function, a yeast two-hybrid screen was performed by using a rat brain cDNA library. This screen identified a unique clone of 1.4 kb, which encoded a 79-aa brain-specific protein that bound the catalytic domain of CaM-KII alpha and beta and potently inhibited kinase activity with an IC50 of 50 nM. The inhibitory protein (CaM-KIIN), and a 28-residue peptide derived from it (CaM-KIINtide), was highly selective for inhibition of CaM-KII with little effect on CaM-KI, CaM-KIV, CaM-KK, protein kinase A, or protein kinase C. CaM-KIIN interacted only with activated CaM-KII (i. e., in the presence of Ca2+/CaM or after autophosphorylation) by using glutathione S-transferase/CaM-KIIN precipitations as well as coimmunoprecipitations from rat brain extracts or from HEK293 cells cotransfected with both constructs. Colocalization of CaM-KIIN with activated CaM KII was demonstrated in COS-7 cells transfected with green fluorescent protein fused to CaM-KIIN. In COS-7 cells phosphorylation of transfected alpha-amino-3 hydroxy-5-methyl-4-isoxazolepropionic acid-type glutamate receptors by CaM-KII, but not by protein kinase C, was blocked upon cotransfection with CaM-KIIN. These results characterize a potent and specific cellular inhibitor of CaM-KII that may have an important role in the physiological regulation of this key protein kinase. PMID- 9724801 TI - Role for neuronally derived fractalkine in mediating interactions between neurons and CX3CR1-expressing microglia. AB - A recently identified chemokine, fractalkine, is a member of the chemokine gene family, which consists principally of secreted, proinflammatory molecules. Fractalkine is distinguished structurally by the presence of a CX3C motif as well as transmembrane spanning and mucin-like domains and shows atypical constitutive expression in a number of nonhematopoietic tissues, including brain. We undertook an extensive characterization of this chemokine and its receptor CX3CR1 in the brain to gain insights into use of chemokine-dependent systems in the central nervous system. Expression of fractalkine in rat brain was found to be widespread and localized principally to neurons. Recombinant rat CX3CR1, as expressed in Chinese hamster ovary cells, specifically bound fractalkine and signaled in the presence of either membrane-anchored or soluble forms of fractalkine protein. Fractalkine stimulated chemotaxis and elevated intracellular calcium levels of microglia; these responses were blocked by anti-CX3CR1 antibodies. After facial motor nerve axotomy, dramatic changes in the levels of CX3CR1 and fractalkine in the facial nucleus were evident. These included increases in the number and perineuronal location of CX3CR1-expressing microglia, decreased levels of motor neuron-expressed fractalkine mRNA, and an alteration in the forms of fractalkine protein expressed. These data describe mechanisms of cellular communication between neurons and microglia, involving fractalkine and CX3CR1, which occur in both normal and pathological states of the central nervous system. PMID- 9724802 TI - Mental chronometry using latency-resolved functional MRI. AB - Vascular responses to neural activity are exploited as the basis of a number of brain imaging techniques. The vascular response is thought to be too slow to resolve the temporal sequence of events involved in cognitive tasks, and hence, imaging studies of mental chronometry have relied on techniques such as the evoked potential. Using rapid functional MRI (fMRI) of single trials of two simple behavioral tasks, we demonstrate that while the microvascular response to the onset of neural activity is delayed consistently by several seconds, the relative timing between the onset of the fMRI responses in different brain areas appears preserved. We examined a number of parameters that characterize the fMRI response and determined that its onset time is best defined by the inflection point from the resting baseline. We have found that fMRI onset latencies determined in this manner correlate well with independently measurable parameters of the tasks such as reaction time or stimulus presentation time and can be used to determine the origin of processing delays during cognitive or perceptual tasks with a temporal accuracy of tens of milliseconds and spatial resolution of millimeters. PMID- 9724803 TI - Selectively enhanced contextual fear conditioning in mice lacking the transcriptional regulator CCAAT/enhancer binding protein delta. AB - CCAAT/enhancer binding protein delta (C/EBPdelta) is a transcriptional regulator implicated in the hepatic acute phase response and in adipogenic and myeloid cell differentiation. We found that C/EBPdelta is widely expressed in the peripheral and central nervous systems, including neurons of the hippocampal formation, indicating a role in neural functions. To examine the role of C/EBPdelta in vivo, we generated mice with a targeted deletion of the C/EBPdelta gene. This mutation does not interfere with normal embryonic and postnatal development. Performance in a battery of behavioral tests indicates that basic neurological functions are normal. Furthermore, performance in a Morris water maze task suggests that C/EBPdelta mutant mice have normal spatial learning. However, in the contextual and auditory-cue-conditioned fear task, C/EBPdelta null mice displayed significantly more conditioned freezing to the test context than did wild-type controls, but equivalent conditioning to the auditory cue. These data demonstrate a selectively enhanced contextual fear response in mice carrying a targeted genomic mutation and implicate C/EBPdelta in the regulation of a specific type of learning and memory. PMID- 9724804 TI - Isoform-specific effects of human apolipoprotein E on brain function revealed in ApoE knockout mice: increased susceptibility of females. AB - Apolipoprotein E (apoE) mediates the redistribution of lipids among cells and is expressed at highest levels in brain and liver. Human apoE exists in three major isoforms encoded by distinct alleles (epsilon2, epsilon3, and epsilon4). Compared with APOE epsilon2 and epsilon3, APOE epsilon4 increases the risk of cognitive impairments, lowers the age of onset of Alzheimer's disease (AD), and decreases the response to AD treatments. Besides age, inheritance of the APOE epsilon4 allele is the most important known risk factor for the development of sporadic AD, the most common form of this illness. Although numerous hypotheses have been advanced, it remains unclear how APOE epsilon4 might affect cognition and increase AD risk. To assess the effects of distinct human apoE isoforms on the brain, we have used the neuron-specific enolase (NSE) promoter to express human apoE3 or apoE4 at similar levels in neurons of transgenic mice lacking endogenous mouse apoE. Compared with NSE-apoE3 mice and wild-type controls, NSE-apoE4 mice showed impairments in learning a water maze task and in vertical exploratory behavior that increased with age and were seen primarily in females. These findings demonstrate that human apoE isoforms have differential effects on brain function in vivo and that the susceptibility to apoE4-induced deficits is critically influenced by age and gender. These results could be pertinent to cognitive impairments observed in human APOE epsilon4 carriers. NSE-apoE mice and similar models may facilitate the preclinical assessment of treatments for apoE related cognitive deficits. PMID- 9724805 TI - beta-adrenergic receptor-induced activation of nerve growth factor gene transcription in rat cerebral cortex involves CCAAT/enhancer-binding protein delta. AB - Stimulation of beta-adrenergic receptors (BAR) by clenbuterol (CLE) increases nerve growth factor (NGF) biosynthesis in the rat cerebral cortex but not in other regions of the brain. We have explored the transcription mechanisms that may account for the cortex-specific activation of the NGF gene. Although the NGF promoter contains an AP-1 element, AP-1-binding activity in the cerebral cortex was not induced by CLE, suggesting that other transcription factors govern the brain area-specific induction of NGF. Because BAR activation increases cAMP levels, we examined the role of CCAAT/enhancer-binding proteins (C/EBP), some of which are known to be cAMP-inducible. In C6-2B glioma cells, whose NGF expression is induced by BAR agonists, (i) CLE increased C/EBPdelta-binding activity, (ii) NGF mRNA levels were increased by overexpressing C/EBPdelta, and (iii) C/EBPdelta increased the activity of an NGF promoter-reporter construct. Moreover, DNase footprinting and deletion analyses identified a C/EBPdelta site in the proximal region of the NGF promoter. C/EBPdelta appears to be responsible for the BAR mediated activation of the NGF gene in vivo, since CLE elicited a time-dependent increase in C/EBPdelta-binding activity in the cerebral cortex only. Our data suggest that, while AP-1 may regulate basal levels of NGF expression, C/EBPdelta is a critical component determining the area-specific expression of NGF in response to BAR stimulation. PMID- 9724806 TI - Nerve growth factor rapidly suppresses basal, NMDA-evoked, and AMPA-evoked nitric oxide synthase activity in rat hippocampus in vivo. AB - In adult forebrain, nerve growth factor (NGF) influences neuronal maintenance and axon sprouting and is neuroprotective in several injury models through mechanisms that are incompletely understood. Most NGF signaling is thought to occur after internalization and retrograde transport of trkA receptor and be mediated through the nucleus. However, NGF expression in hippocampus is rapidly and sensitively regulated by synaptic activity, suggesting that NGF exerts local effects more dynamically than possible through signaling requiring retrograde transport to distant afferent neurons. Interactions have been reported between NGF and nitric oxide (NO). Because NO affects both neural plasticity and degeneration, and trk receptors can mediate signaling within minutes, we hypothesized that NGF might rapidly modulate NO production. Using in vivo microdialysis we measured conversion of L-[14C]arginine to L-[14C]citrulline as an accurate reflection of NO synthase (NOS) activity in adult rat hippocampus. NGF significantly reduced NOS activity to 61% of basal levels within 20 min of onset of delivery and maintained NOS activity at less than 50% of baseline throughout 3 hr of delivery. This effect did not occur with control protein (cytochrome c) and was not mediated by an effect of NGF on glutamate levels. In addition, simultaneous delivery of NGF prevented significant increases in NOS activity triggered by the glutamate receptor agonists N-methyl-D-aspartate (NMDA) and alpha-amino-3-hydroxy 5-methyl-4-isoxazolepropionic acid (AMPA). Rapid suppression by NGF of basal and glutamate-stimulated NOS activity may regulate neuromodulatory functions of NO or protect neurons from NO toxicity and suggests a novel mechanism for rapidly mediating functions of NGF and other neurotrophins. PMID- 9724807 TI - How the owl resolves auditory coding ambiguity. AB - The barn owl (Tyto alba) uses interaural time difference (ITD) cues to localize sounds in the horizontal plane. Low-order binaural auditory neurons with sharp frequency tuning act as narrow-band coincidence detectors; such neurons respond equally well to sounds with a particular ITD and its phase equivalents and are said to be phase ambiguous. Higher-order neurons with broad frequency tuning are unambiguously selective for single ITDs in response to broad-band sounds and show little or no response to phase equivalents. Selectivity for single ITDs is thought to arise from the convergence of parallel, narrow-band frequency channels that originate in the cochlea. ITD tuning to variable bandwidth stimuli was measured in higher-order neurons of the owl's inferior colliculus to examine the rules that govern the relationship between frequency channel convergence and the resolution of phase ambiguity. Ambiguity decreased as stimulus bandwidth increased, reaching a minimum at 2-3 kHz. Two independent mechanisms appear to contribute to the elimination of ambiguity: one suppressive and one facilitative. The integration of information carried by parallel, distributed processing channels is a common theme of sensory processing that spans both modality and species boundaries. The principles underlying the resolution of phase ambiguity and frequency channel convergence in the owl may have implications for other sensory systems, such as electrolocation in electric fish and the computation of binocular disparity in the avian and mammalian visual systems. PMID- 9724808 TI - Modulation of a calcium-sensitive nonspecific cation channel by closely associated protein kinase and phosphatase activities. AB - Regulation of nonspecific cation channels often underlies neuronal bursting and other prolonged changes in neuronal activity. In bag cell neurons of Aplysia, it recently has been suggested that an intracellular messenger-induced increase in the activity of a nonspecific cation channel may underlie the onset of a 30-min period of spontaneous action potentials referred to as the "afterdischarge. " In patch clamp studies of the channel, we show that the open probability of the channel can be increased by an average of 10. 7-fold by application of ATP to the cytoplasmic side of patches. Duration histograms indicate that the increase is primarily a result of a reduction in the duration and percentage of channel closures described by the slowest time constant. The increase in open probability was not observed using 5'-adenylylimidodiphosphate, a nonhydrolyzable ATP analog, and was blocked in the presence of H7 or the more specific calcium/phospholipid dependent protein kinase C (PKC) inhibitor peptide(19-36). Because the increase in activity observed in response to ATP occurred without application of protein kinase, our results indicate that a kinase endogenous to excised patches mediates the effect. The effect of ATP could be reversed by exogenously applied protein phosphatase 1 or by a microcystin-sensitive phosphatase also endogenous to excised patches. These results, together with work demonstrating the presence of a protein tyrosine phosphatase in these patches, suggest that the cation channel is part of a regulatory complex including at least three enzymes. This complex may act as a molecular switch to activate the cation channel and, thereby, trigger the afterdischarge. PMID- 9724809 TI - A molecular mechanism for signaling between seven-transmembrane receptors: evidence for a redistribution of G proteins. AB - Although activation of one seven-transmembrane receptor can influence the response of a separate seven-transmembrane receptor, e. g., the phenomenon of synergism, the underlying mechanism(s) for this signaling process is unclear. The present study investigated communication between two receptors that exhibit classical synergism, e.g., human platelet thrombin and thromboxane A2 receptors. Activation of thrombin receptors caused an increase in ligand affinity of thromboxane A2 receptors. This effect (i) was shown to be specific, since a similar increase in ligand affinity was not caused by ADP or A23187; (ii) did not require cytosolic components, e.g., kinases, proteases, phosphatases, etc., because it occurred in isolated platelet membranes; (iii) was G protein-mediated because it was blocked by an Galphaq C terminus antibody; and (iv) was associated with a net increase in Galphaq coupling to thromboxane A2 receptors. Collectively, these data provide evidence that seven-transmembrane receptors that share a common Galpha subunit can communicate with each other via a redistribution of their G proteins. Thus, activation of thrombin receptors increases Galphaq association with thromboxane A2 receptors thereby shifting them to a higher affinity state. This signaling phenomenon, which modulates receptor ligand affinity, may serve as a molecular mechanism for cellular adaptive processes such as synergism. PMID- 9724810 TI - Nociceptin/orphanin FQ-induced nociceptive responses through substance P release from peripheral nerve endings in mice. AB - We have studied the in vivo signaling mechanisms involved in nociceptin/orphanin FQ (Noci)-induced pain responses by using a flexor-reflex paradigm. Noci was 10,000 times more potent than substance P (SP) in eliciting flexor responses after intraplantar injection into the hind limb of mice, but the action of Noci seems to be mediated by SP. Mice pretreated with an NK1 tachykinin receptor antagonist or capsaicin, or mice with a targeted disruption of the tachykinin 1 gene no longer respond to Noci. The action of Noci appears to be mediated by the Noci receptor, a pertussis toxin-sensitive G protein-coupled receptor that stimulates inositol trisphosphate receptor and Ca2+ influx. These findings suggest that Noci indirectly stimulates nerve endings of nociceptive primary afferent neurons through a local SP release. PMID- 9724811 TI - Cyclooxygenase-2 inhibition prevents delayed death of CA1 hippocampal neurons following global ischemia. AB - The inducible isoform of the enzyme cyclooxygenase-2 (COX2) is an immediate early gene induced by synaptic activity in the brain. COX2 activity is an important mediator of inflammation, but it is not known whether COX2 activity is pathogenic in brain. To study the role of COX2 activity in ischemic injury in brain, expression of COX2 mRNA and protein and the effect of treatment with a COX2 inhibitor on neuronal survival in a rat model of global ischemia were determined. Expression of both COX2 mRNA and protein was increased after ischemia in CA1 hippocampal neurons before their death. There was increased survival of CA1 neurons in rats treated with the COX2-selective inhibitor SC58125 [1-[(4 methylsulfonyl) phenyl]-3-trifluoro-methyl-5-[(4-fluoro)phenyl] pyrazole] before or after global ischemia compared with vehicle controls. Furthermore, hippocampal prostaglandin E2 concentrations 24 h after global ischemia were decreased in drug treated animals compared with vehicle-treated controls. These results suggest that COX2 activity contributes to CA1 neuronal death after global ischemia. PMID- 9724813 TI - Interaction between inducible nitric oxide synthase and cyclooxygenase-2 after cerebral ischemia. AB - Focal cerebral ischemia is associated with expression of both inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), enzymes whose reaction products contribute to the evolution of ischemic brain injury. We tested the hypothesis that, after cerebral ischemia, nitric oxide (NO) produced by iNOS enhances COX-2 activity, thereby increasing the toxic potential of this enzyme. Cerebral ischemia was produced by middle cerebral artery occlusion in rats or mice. Twenty-four hours after ischemia in rats, iNOS-immunoreactive neutrophils were observed in close proximity (<20 micrometer) to COX-2-positive cells at the periphery of the infarct. In the olfactory bulb, only COX-2 positive cells were observed. Cerebral ischemia increased the concentration of the COX-2 reaction product prostaglandin E2 (PGE2) in the ischemic area and in the ipsilateral olfactory bulb. The iNOS inhibitor aminoguanidine reduced PGE2 concentration in the infarct, where both iNOS and COX-2 were expressed, but not in the olfactory bulb, where only COX-2 was expressed. Postischemic PGE2 accumulation was reduced significantly in iNOS null mice compared with wild-type controls (C57BL/6 or SV129). The data provide evidence that NO produced by iNOS influences COX-2 activity after focal cerebral ischemia. Pro-inflammatory prostanoids and reactive oxygen species produced by COX-2 may be a previously unrecognized factor by which NO contributes to ischemic brain injury. The pathogenic effect of the interaction between NO, or a derived specie, and COX-2 is likely to play a role also in other brain diseases associated with inflammation. PMID- 9724812 TI - ZK200775: a phosphonate quinoxalinedione AMPA antagonist for neuroprotection in stroke and trauma. AB - Stroke and head trauma are worldwide public health problems and leading causes of death and disability in humans, yet, no adequate neuroprotective treatment is available for therapy. Glutamate antagonists are considered major drug candidates for neuroprotection in stroke and trauma. However, N-methyl-D-aspartate antagonists failed clinical trials because of unacceptable side effects and short therapeutic time window. alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) antagonists derived from the quinoxalinedione scaffold cannot be used in humans because of their insolubility and resulting renal toxicity. Therefore, achieving water solubility of quinoxalinediones without loss of selectivity and potency profiles becomes a major challenge for medicinal chemistry. One of the major tenets in the chemistry of glutamate antagonists is that the incorporation of phosphonate into the glutamate framework results in preferential N-methyl-D aspartate antagonism. Therefore, synthesis of phosphonate derivatives of quinoxalinediones was not pursued because of a predicted loss of their selectivity toward AMPA. Here, we report that introduction of a methylphosphonate group into the quinoxalinedione skeleton leaves potency as AMPA antagonists and selectivity for the AMPA receptor unchanged and dramatically improves solubility. One such novel phosphonate quinoxalinedione derivative and competitive AMPA antagonist ZK200775 exhibited a surprisingly long therapeutic time window of >4 h after permanent occlusion of the middle cerebral artery in rats and was devoid of renal toxicity. Furthermore, delayed treatment with ZK200775 commencing 2 h after onset of reperfusion in transient middle cerebral artery occlusion resulted in a dramatic reduction of the infarct size. ZK200775 alleviated also both cortical and hippocampal damage induced by head trauma in the rat. These observations suggest that phosphonate quinoxalinedione-based AMPA antagonists may offer new prospects for treatment of stroke and trauma in humans. PMID- 9724814 TI - Syntaxin 1A inhibits CFTR chloride channels by means of domain-specific protein protein interactions. AB - Previously we showed that the functional activity of the epithelial chloride channel that is encoded by the cystic fibrosis gene (CFTR) is reciprocally modulated by two components of the vesicle fusion machinery, syntaxin 1A and Munc 18. Here we report that syntaxin 1A inhibits CFTR chloride channels by means of direct and domain-specific protein-protein interactions. Syntaxin 1A stoichiometrically binds to the N-terminal cytoplasmic tail of CFTR, and this binding is blocked by Munc-18. The modulation of CFTR currents by syntaxin 1A is eliminated either by deletion of this tail or by injecting this tail as a blocking peptide into coexpressing Xenopus oocytes. The CFTR binding site on syntaxin 1A maps to the third predicted helical domain (H3) of this membrane protein. Moreover, CFTR Cl- currents are effectively inhibited by a minimal syntaxin 1A construct (i.e., the membrane-anchored H3 domain) that cannot fully substitute for wild-type syntaxin 1A in membrane fusion reactions. We also show that syntaxin 1A binds to and inhibits the activities of disease-associated mutants of CFTR, and that the chloride current activity of recombinant DeltaF508 CFTR (i.e., the most common cystic fibrosis mutant) can be potentiated by disrupting its interaction with syntaxin 1A in cultured epithelial cells. Our results provide evidence for a direct physical interaction between CFTR and syntaxin 1A that limits the functional activities of normal and disease associated forms of this chloride channel. PMID- 9724815 TI - The negative feedback actions of progesterone on gonadotropin-releasing hormone secretion are transduced by the classical progesterone receptor. AB - Progesterone (P) powerfully inhibits gonadotropin-releasing hormone (GnRH) secretion in ewes, as in other species, but the neural mechanisms underlying this effect remain poorly understood. Using an estrogen (E)-free ovine model, we investigated the immediate GnRH and luteinizing hormone (LH) response to acute manipulations of circulating P concentrations and whether this response was mediated by the nuclear P receptor. Simultaneous hypophyseal portal and jugular blood samples were collected over 36 hr: 0-12 hr, in the presence of exogenous P (P treatment begun 8 days earlier); 12-24 hr, P implant removed; 24-36 hr, P implant reinserted. P removal caused a significant rapid increase in the GnRH pulse frequency, which was detectable within two pulses (175 min). P insertion suppressed the GnRH pulse frequency even faster: the effect detectable within one pulse (49 min). LH pulsatility was modulated identically. The next two experiments demonstrated that these effects of P are mediated by the nuclear P receptor since intracerebroventricularly infused P suppressed LH release but 3alpha-hydroxy-5alpha-pregnan-20-one, which operates through the type A gamma aminobutyric acid receptor, was without effect and pretreatment with the P receptor antagonist RU486 blocked the ability of P to inhibit LH. Our final study showed that P exerts its acute suppression of GnRH through an E-dependent system because the effects of P on LH secretion, lost after long-term E deprivation, are restored after 2 weeks of E treatment. Thus we demonstrate that P acutely inhibits GnRH through an E-dependent nuclear P-receptor system. PMID- 9724816 TI - Two-dimensional confocal images of organization, density, and gating of focal Ca2+ release sites in rat cardiac myocytes. AB - In cardiac myocytes Ca2+ cross-signaling between Ca2+ channels and ryanodine receptors takes place by exchange of Ca2+ signals in microdomains surrounding dyadic junctions, allowing first the activation and then the inactivation of the two Ca2+-transporting proteins. To explore the details of Ca2+ signaling between the two sets of receptors we measured the two-dimensional cellular distribution of Ca2+ at 240 Hz by using a novel confocal imaging technique. Ca2+ channel triggered Ca2+ transients could be resolved into dynamic "Ca2+ stripes" composed of hundreds of discrete focal Ca2+ releases, appearing as bright fluorescence spots (radius congruent with 0.5 micrometer) at reproducible sites, which often coincided with t-tubules as visualized with fluorescent staining of the cell membrane. Focal Ca2+ releases triggered stochastically by Ca2+ current (ICa) changed little in duration ( congruent with7 ms) and size (congruent with100,000 Ca ions) between -40 and +60 mV, but their frequency of activation and first latency mirrored the kinetics and voltage dependence of ICa. The resolution of 0.95 +/- 0. 13 reproducible focal Ca2+ release sites per micrometer3 in highly Ca2+-buffered cells, where diffusion of Ca2+ is limited to 50 nm, suggests the presence of about one independent, functional Ca2+ release site per half sarcomere. The density and distribution of Ca2+ release sites suggest they correspond to dyadic junctions. The abrupt onset and termination of focal Ca2+ releases indicate that the cluster of ryanodine receptors in individual dyadic junctions may operate in a coordinated fashion. PMID- 9724818 TI - Transport of uncharged organic solutes in Xenopus oocytes expressing red cell anion exchangers (AE1s). AB - When expressed in Xenopus oocytes, the trout red cell anion exchanger tAE1, but not the mouse exchanger mAE1, elicited a transport of electroneutral solutes (sorbitol, urea) in addition to the expected anion exchange activity. Chimeras constructed from mAE1 and tAE1 allowed us to identify the tAE1 domains involved in the induction of these transports. Expression of tAE1 (but not mAE1) is known to generate an anion conductance associated with a taurine transport. The present data provide evidence that (i) the capacity of tAE1 and tAE1 chimeras to generate urea and sorbitol permeability also was associated with an anion conductance; (ii) the same inhibitors affected both the permeability of solutes and anion conductance; and (iii) no measurable water transport was associated with the tAE1 dependent conductance. These results support the view that fish red blood cells, to achieve cell volume regulation in response to hypotonic swelling, activate a tAE1-associated anion channel that can mediate the passive transport of taurine and electroneutral solutes. PMID- 9724817 TI - Oocytes are a source of catecholamines in the primate ovary: evidence for a cell cell regulatory loop. AB - Catecholamines, thought to derive from the extrinsic innervation of the ovary, participate in the regulation of ovarian development and mature gonadal function. Recently, intraovarian neurons containing tyrosine hydroxylase (TH), the rate limiting enzyme in catecholamine biosynthesis, were described in the ovary of nonhuman primates. We now show that the primate ovary expresses both the genes encoding TH and dopamine beta-hydroxylase (DBH), the key enzymes in norepinephrine (NE) biosynthesis. Ovarian neurons were identified as a site of TH and DBH gene expression, and surprisingly, oocytes were identified as an exclusive site of DBH synthesis. Oocytes contain neither TH mRNA nor protein, indicating that they are unable to synthesize dopamine (DA). They did, however, express a DA transporter gene identical to that found in human brain. The physiological relevance of this transporter system and DBH in oocytes was indicated by the ability of isolated oocytes to metabolize exogenous DA into NE. Isolated follicles containing oocytes-but not those from which the oocytes had been removed-responded to DA with an elevation in cAMP levels; this elevation was prevented by propranolol, a beta-adrenoreceptor antagonist. The results suggest that oocytes and somatic cells are linked by a neuroendocrine loop consisting of NE synthesized in oocytes from actively transported DA and cAMP produced by somatic follicular cells in response to NE-induced beta-adrenoreceptor activation. PMID- 9724819 TI - In vivo responses of the primate corpus luteum to luteinizing hormone and chorionic gonadotropin. AB - Although it is well established that the secretory activity of the corpus luteum absolutely depends on the presence of pituitary-derived luteinizing hormone (LH), it is unknown why the life span of the corpus luteum is extended during early pregnancy by the placental production of chorionic gonadotropin (CG) but regresses in the presence of LH despite the fact that CG and LH have similar actions on the corpus luteum. To compare the responses of the corpus luteum to LH and human CG (hCG), cynomolgus monkeys whose endogenous gonadotropin secretion was blocked during the luteal phase of the menstrual cycle with a gonadotropin releasing hormone antagonist were i.v. infused with either LH or CG. Infusion of LH at a constant rate overcame the gonadotropin-releasing hormone antagonist mediated premature luteal regression but failed to prolong the functional life span of the corpus luteum. Continuous infusions of hCG did not effect a pregnancy like pattern of gonadotropin secretion, but the functional life span of the corpus luteun was extended in two of three animals. Infusion of either LH or hCG in an exponentially increasing manner prolonged the functional life span of the corpus luteum beyond its normal duration. These results indicate that luteal regression at the termination of nonfertile menstrual cycles is caused by a large reduction in the responsiveness of the aging corpus luteum to LH, which can be overcome by elevated concentrations of either LH or CG. PMID- 9724820 TI - A response regulator of cyanobacteria integrates diverse environmental signals and is critical for survival under extreme conditions. AB - Microorganisms must sense their environment and rapidly tune their metabolism to ambient conditions to efficiently use available resources. We have identified a gene encoding a response regulator, NblR, that complements a cyanobacterial mutant unable to degrade its light-harvesting complex (phycobilisome), in response to nutrient deprivation. Cells of the nblR mutant (i) have more phycobilisomes than wild-type cells during nutrient-replete growth, (ii) do not degrade phycobilisomes during sulfur, nitrogen, or phosphorus limitation, (iii) cannot properly modulate the phycobilisome level during exposure to high light, and (iv) die rapidly when starved for either sulfur or nitrogen, or when exposed to high light. Apart from regulation of phycobilisome degradation, NblR modulates additional functions critical for cell survival during nutrient-limited and high light conditions. NblR does not appear to be involved in acclimation responses that occur only during a specific nutrient limitation. In contrast, it controls at least some of the general acclimation responses; those that occur during any of a number of different stress conditions. NblR plays a pivotal role in integrating different environmental signals that link the metabolism of the cell to light harvesting capabilities and the activities of the photosynthetic apparatus; this modulation is critical for cell survival. PMID- 9724823 TI - Editorial PMID- 9724821 TI - A single gene of chloroplast origin codes for mitochondrial and chloroplastic methionyl-tRNA synthetase in Arabidopsis thaliana. AB - One-fifth of the tRNAs used in plant mitochondrial translation is coded for by chloroplast-derived tRNA genes. To understand how aminoacyl-tRNA synthetases have adapted to the presence of these tRNAs in mitochondria, we have cloned an Arabidopsis thaliana cDNA coding for a methionyl-tRNA synthetase. This enzyme was chosen because chloroplast-like elongator tRNAMet genes have been described in several plant species, including A. thaliana. We demonstrate here that the isolated cDNA codes for both the chloroplastic and the mitochondrial methionyl tRNA synthetase (MetRS). The protein is transported into isolated chloroplasts and mitochondria and is processed to its mature form in both organelles. Transient expression assays using the green fluorescent protein demonstrated that the N-terminal region of the MetRS is sufficient to address the protein to both chloroplasts and mitochondria. Moreover, characterization of MetRS activities from mitochondria and chloroplasts of pea showed that only one MetRS activity exists in each organelle and that both are indistinguishable by their behavior on ion exchange and hydrophobic chromatographies. The high degree of sequence similarity between A. thaliana and Synechocystis MetRS strongly suggests that the A. thaliana MetRS gene described here is of chloroplast origin. PMID- 9724822 TI - Protein kinase CK2 interacts with and phosphorylates the Arabidopsis circadian clock-associated 1 protein. AB - The circadian clock-associated 1 (CCA1) gene encodes a Myb-related transcription factor that has been shown to be involved in the phytochrome regulation of Lhcb1*3 gene expression and in the function of the circadian oscillator in Arabidopsis thaliana. By using a yeast interaction screen to identify proteins that interact with CCA1, we have isolated a cDNA clone encoding a regulatory (beta) subunit of the protein kinase CK2 and have designated it as CKB3. CKB3 is the only reported example of a third beta-subunit of CK2 found in any organism. CKB3 interacts specifically with CCA1 both in a yeast two-hybrid system and in an in vitro interaction assay. Other subunits of CK2 also show an interaction with CCA1 in vitro. CK2 beta-subunits stimulate binding of CCA1 to the CCA1 binding site on the Lhcb1*3 gene promoter, and recombinant CK2 is able to phosphorylate CCA1 in vitro. Furthermore, Arabidopsis plant extracts contain a CK2-like activity that affects the formation of a DNA-protein complex containing CCA1. These results suggest that CK2 can modulate CCA1 activity both by direct interaction and by phosphorylation of the CCA1 protein and that CK2 may play a role in the function of CCA1 in vivo. PMID- 9724824 TI - Potential benefits of resistance exercise training on nutritional status in renal failure. AB - Resistance or strength exercise training may help reverse the malnutrition common among patients in chronic renal failure and delay the progression of renal disease. Resistance training is characterized by resisting, lifting, and lowering weights. It results in muscle mass accretion, improved physical function, and slowed progression of muscle wasting. Resistance exercise training for a period of 8 to 12 weeks results in significant increases in muscle mass, muscle strength, and muscle function in frail "healthy" elderly individuals as well as in specific patient populations. States of malnutrition leading to muscle wasting directly affect lean tissue mass and functional capacity. Even at dietary protein intake below the Recommended Dietary Allowances, resistance training appears to exert an anabolic effect by improving energy intake and protein use allowing nitrogen retention. The potential benefits of resistance exercise extend beyond this direct impact on protein metabolism. They include improvements in functional capacity such as gait, balance, mobility, strength, exercise tolerance, improved glucose uptake, insulin sensitivity, and self-efficacy and self-esteem. Currently, the effects of resistance exercise in renal patients are unknown, although they are well shown in the case of other diseases. The potential benefits that resistance exercise training may have on muscle mass and function, nutritional status, hyperglycemia, disease progression, and the overall mental well-being of renal patients deserve further investigation. As an adjunct to current treatment modalities for chronic renal failure, resistance exercise may serve as a cost-effective, interdisciplinary, noninvasive approach to counteract malnutrition and improve the quality of life. PMID- 9724825 TI - The hemodialysis pilot study: nutrition program and participant characteristics at baseline. The HEMO Study Group. AB - OBJECTIVE: Describe the nutrition program (assessments and interventions) and the participants' baseline nutritional characteristics in the Hemodialysis Pilot Study. DESIGN: Cross sectional survey in which hemodialysis patients were examined during 10 weeks of baseline (BL), before randomization study interventions (dose and flux). SETTING: Four hemodialysis centers (eight dialysis units in total). PATIENTS: Twenty-nine male (mean age, 63 years; range, 34 to 75) and 20 female (mean age, 61 years; range, 29 to 73) hemodialysis patients. INTERVENTIONS: None during BL. MAIN OUTCOME MEASURES: Feasibility of implementing the proposed nutrition program before conducting the full-scale trial, and description of baseline characteristics related to nutrition. RESULTS: A nutrition program was developed to assess nutritional status during BL and follow up periods and to intervene in patients with weight loss or decreasing serum albumin. Methods for collecting biochemical, dietary and anthropometric data were implemented at four clinical centers. At baseline, mean protein intake estimated by single pool normalized protein catabolic rate was 0.95 +/- 0.21 gm/kg adjusted body weight (ABW) (n = 42) and by diet record assisted recalls (n = 47) 0.94 +/- 0.36 gm/kg ABW/d, respectively. Mean energy intake was 22.8 +/- 8 kcal/kg ABW/day (n = 39). Mean serum albumin concentration using the bromcresol green method was 3.8 +/- 0.4 gm/dL (n = 40). Mean body mass index was within the normal limits of 19-27 kg/m2. Mean skinfold thicknesses in females, but not males, were shifted toward the lower end of usual distributions for healthy individuals. CONCLUSIONS: The goal of designing, developing, and implementing the diet and nutrition component, and related data collection for the HEMO pilot study was accomplished at four separate clinical centers. Baseline mean protein and energy intake were low, suggesting that continuing dietary surveillance is needed. The ongoing full scale HEMO study will provide the first prospective analysis of dietary intake, nutritional status, and outcome in maintenance hemodialysis patients as a function of dialysis dose and membrane flux. PMID- 9724826 TI - Using the hemodialysis prognostic nutrition index and urea reduction ratio to predict morbidity and mortality: a pilot study of the 1995 council on renal nutrition national research question. AB - OBJECTIVE: To validate the use of the hemodialysis prognostic nutrition index (HPNI) in an alternate hemodialysis population and to determine if use of urea reduction ratio would improve use in outcome prediction for morbidity and mortality. DESIGN: Prospective random cohort. SETTING: Hospital based non-for profit outpatient dialysis unit. PATIENTS: Forty chronic hemodialysis patients, 50% men, 50% black, 16% diabetic, 67.2 mean months on hemodialysis, mean age 54.5 years. INTERVENTIONS: None; observational; tracking of routinely collected demographic, biochemical, and clinical data. MAIN OUTCOME MEASURES: Number of times and days hospitalized, mortality RESULTS: Plotting of HPNI against urea reduction ratio produced risk quadrants for hospitalization that were more predictive than HPNI alone. CONCLUSION: Application continues as a multicenter collaborative Council on Renal Nutrition National Research Question. PMID- 9724827 TI - Safety and tolerance of medical nutritional products as sole sources of nutrition in people on hemodialysis. AB - OBJECTIVE: Establish and compare the safety and tolerance of three medical nutritional products when used as sole sources of nutrition in stable hemodialysis patients. DESIGN: Prospectively randomized, controlled, single blind, parallel design. SETTING: Three outpatient hemodialysis clinics. PARTICIPANTS: Seventy-nine normally nourished, stable, anuric, adequately dialyzed, adult outpatients with end-stage renal disease (ESRD) and requiring thrice weekly hemodialysis. INTERVENTION: A 3-week trial was conducted. During the first week, baseline medical history and physical examination, gastrointestinal symptom, urea kinetic, bowel habit, and biochemical data were collected while participants ingested their usual diet. During the last 2 weeks, the same data were collected while participants orally ingested 35 kcal/kg actual weight/d of one of three medical nutritional products as a sole source of nutrition. Products were a standard medical nutritional (EN-9527) and two renal nutritionals (EN-9528 and EN-9529). The latter product was a reformulation of EN 9528 and contained added beta-carotene and fructooligosaccharides. MAIN OUTCOME MEASURES: Gastrointestinal symptoms, bowel habits (stool frequency and consistency), routine blood chemistries, urea kinetics, and normalized protein catabolic rate (nPCR) RESULTS: All three groups achieved a mean energy and protein intake of approximately 35 kcal/kg/d and 1.25 g protein/kg/d during the last 10 days of the sole source feeding period. Adherence with the formula ingestion targets was assessed using both a patient-completed product consumption log and nPCR. By intent to treat analysis, there were no changes in number or severity of gastrointestinal symptoms, stool frequency or stool consistency, or urea kinetics between the baseline week and during product consumption. In comparison to the standard formulation, the disease-specific formulations resulted in improved serum phosphorus and calcium-phosphorus product. Patients receiving the fructooligosaccharide-containing product (EN-9529), by Chi-squared analysis, had less constipation than for the comparable product without oligosaccharides (EN-9528) or the standard medical nutritional (EN-9527). CONCLUSION: Use of enteral nutritionals as a sole source of nutrition is both possible and well tolerated in hemodialyzed patients. Selection of a disease specific formulation offered advantages over a standard formulation in the management of biochemical complications of renal disease when the products were used as a sole source of nutrition. PMID- 9724828 TI - The relationship between plasma homocysteine and amino acid concentrations in patients with end-stage renal disease. AB - OBJECTIVE: Observe the relationship between homocysteine and other amino acids in patients with end-stage renal disease (ESRD). DESIGN: A cross-sectional study of amino acids and homocysteine and comparison of the correlations between ESRD and control group. SETTING: Nephrology unit of Soonchunhyang University hospital in Chunan, Korea. PARTICIPANTS: Forty-five ESRD patients and 30 control volunteers. MAIN OUTCOME MEASURES: Plasma amino acids and homocysteine. RESULTS: Concentrations of asparate, proline and cysteine were higher and serine, tyrosine, valine, isoleuline, leucine, and lysine levels were lower in the ESRD group than in control group. The branched chain amino acids (BCAAs) and essential amino acid were lower in the ESRD group than in the control group, but there was no difference in non-essential amino acid and total amino acid between the two groups. The mean plasma total homocysteine concentrations were 6 +/- 1 mmol/L in the control group and 14 +/- 4 mmol/L in the ESRD group (P < .001). In the ESRD group, homocysteine concentrations showed a direct correlation with the concentration of histidine (R2: .403, P < .001), valine (R2: .324, P < .01), leucine (R2: .400, P < .01), isoleucine (R2: .351, P < .005), cysteine (R2: .287, P < .001), methionine (R2: .256, P < .01), BCAA (R2: .50, P < .01), and essential amino acid (R2: .416, P < .01). In the control group, no correlation between the homocysteine and amino acid concentrations was found. CONCLUSIONS: Contrary to the control group, the homocysteine concentrations showed a direct correlation with the concentration of valine, leucine, isoleucine, methionine, and histidine levels in the ESRD group. Altered essential amino acid metabolism, specifically BCAAs and histidine, influence hyperhomocysteinemia in ESRD. Further study is needed to confirm this theory. PMID- 9724829 TI - Medicinal herb use and the renal patient. AB - Medicinal herb use, although a popular branch of alternative medicine, may be inappropriate for the renal patient. The pharmacological activity, chemical components, and microbial content of herbs, as well as their ability to interfere with prescription medications, make medicinal herbs potentially dangerous for the renal patient. The purpose of this report is to inform the medical professional of the implications of renal patients using medicinal herbs. PMID- 9724830 TI - Make phosphorus education hit home! PMID- 9724833 TI - MEETING ANNONCEMENTS (Educational opportunities) PMID- 9724832 TI - Message from the chairperson PMID- 9724834 TI - 1998 spring clinical meeting call for abstracts PMID- 9724835 TI - Quality of life 10 years after a very severe traumatic brain injury (TBI): the perspective of the injured and the closest relative. AB - This study is a further follow-up of a group of 15 very severely injured TBI patients who have earlier been followed-up 5 years after the injury, and their closest relatives. The aim of this study was to evaluate the factors related to the quality of life of the injured and the strain felt by the relatives. The information was gathered by questionnaires for the injured and the relative and clinical ratings based on the observations of a clinician. The self-reported quality of life of both the injured and their closest relatives was rather high in spite of the various physical, cognitive and emotional/behavioral disturbances. However, the strain felt by many of the relatives was still high 10 years after the injury although it had decreased over the years. The neurobehavioral and emotional disturbances had the most significant effect on the quality of life of the injured and strain felt by the relative. The relationship between the quality of life of the injured and strain felt by the relative was not linear. The implications of the findings for developing different forms of rehabilitation and support systems is discussed. PMID- 9724836 TI - An evaluation of subjective and objective measures of fatigue in patients with brain injury and healthy controls. AB - Three self-report scales and an objective measure were examined for their value in assessing fatigue in patients with brain injury. Patients with brain injury and healthy controls completed the Fatigue Impact Scale (FIS), Visual Analogue Scale for Fatigue (VAS-F) and Fatigue Severity Scale (FSS). Fatigue was objectively measured via a continuous thumb pressing task. Patients scored higher on all fatigue measures than did participants without brain injury. Significant group differences were found on the FIS, the vigour subscale of the VAS-F, and the FSS. The FIS provided a comprehensive assessment of patients' fatigue experience. The FSS, although differentiating between groups, did not provide as comprehensive an examination of fatigue as the FIS and the scale's internal consistency requires review. No significant group differences in fatigue ratings were found on the VAS-F, possibly due to the scale's failure to differentiate between fatigue and sleepiness. The objective measure of fatigue found patients with brain injury fatigued more quickly than participants without brain injury. Although group differences were not significant, this trend suggest that further examination of this fatigue measure is warranted. Overall, patients with brain injury were found to experience significant levels of fatigue and the FIS provided the most comprehensive examination of fatigue. PMID- 9724837 TI - Treatment of aggression and irritability after head injury. AB - Thirteen patients who experienced problems with irritability and aggression following closed head injury (CHI) participated in a non-blind, 8 week open trial and sertraline HCl. Significant reduction in irritability and aggressive outbursts was observed. No significant changes were observed in depressive symptomatology. Results suggest that serotonergic agents may be useful in treating aggression and irritability after head injury. Further placebo controlled studies using serotonergic agents are indicated. PMID- 9724838 TI - Parental report of occurrences and consequences of traumatic brain injury among delinquent and non-delinquent youth. AB - Completed questionnaires from parents of youths attending a public middle school or high school and parents of youths admitted to an institution for juvenile delinquents provided information about incidents of traumatic brain injury (TBI) in their children. Results revealed that approximately 40% of the non-delinquent youth and 50% of the delinquent youth had sustained one or more TBIs during their childhood or youth. The majority of injuries appeared to be mild and had no permanent consequences. However, the parents of more than one-third of the delinquent youth with TBI histories reported long-term effects on academic performance, behavior and emotional control, activity level, and/or interactions with friends and family members; parental reports of long-term effects occurred significantly less frequently among the non-delinquent youth. The most common causes of TBI differed between the two adolescent populations. Non-delinquent youth sustained TBIs most frequently from blows to the head during sporting events, and delinquent youth sustained TBIs with approximately equal frequency from sporting events, fall, motor vehicle accidents and fights. PMID- 9724839 TI - Driving following traumatic brain injury: prevalence, exposure, advice and evaluations. AB - Survivors of traumatic brain injury often have long-term sensory, cognitive and motor deficits that may impair vehicle operation. However, relatively little is known about the driving status and driving characteristics of brain injury survivors. To better understand driving following traumatic brain injury, a survey of driving status, driving exposure, advice received about driving and evaluations of driving competency was administered to a convenience sample of traumatic brain injury survivors (n = 83). The majority of survey participants had experienced either moderate or severe traumatic brain injuries based on the Glasgow Coma Scale. A total of 60% of the survey participants reported that they were currently active drivers. Most individuals (> 60%) who had returned to driving reported driving every day and more than 50 miles per week. Traumatic brain injury survivors frequently received advice about driving from family members, physicians or non-physician health care professionals, but over half (63%) had not been professionally evaluated for driving competency. The presence of high driving exposure, coupled with a lack of widespread driving fitness testing, suggests that some traumatic brain injury survivors have characteristics that may evaluate their risk for vehicle crashes. However, subsequent prospective studies that directly assess driver safety will be needed to confirm this possibility. PMID- 9724840 TI - Gerstmann's syndrome associated with chronic subdural haematoma: a case report. AB - We report a patient who exhibited Gerstmann's syndrome in association with a chronic subdural haematoma. A 71-year-old right-handed woman presented with mild right arm and leg weakness that began 2 weeks prior to admission. Neurological examination on admission revealed a mild right hemiparesis. Neuropsychological examination revealed right-left disorientation, finger agnosia, agraphia, and acalculia, but no language disturbance. A computerized tomographic (CT) scan revealed a large left frontoparietal, extra-axial hypodense fluid collection containing scattered hypodense foci. A left parietal evacuation of the haematoma was performed. Following surgery the patient dramatically improved. We suggest that the direct compression by the chronic subdural haematoma or a hemispheric pressure difference caused Gerstmann's syndrome. This is an unusual report of a Gerstmann's syndrome following chronic subdural haematoma. PMID- 9724841 TI - Medroxyprogesterone in the treatment of aggressive hypersexual behavior in traumatic brain injury. AB - Sexual function is among the many areas affected by traumatic brain injury. The most common change is decreased sexual performance and satisfaction, for the brain injured person and the sexual partner. Hypersexuality, especially inappropriate sexual comments and gestures, is also a common result of traumatic brain injury. A case of hypersexuality in a severely disabled brain injured man is presented. He was successfully treated with medroxyprogesterone acetate after failure of multiple other treatment strategies. The literature is reviewed. An evaluation and treatment strategy for sexual dysfunction post traumatic brain injury is presented. PMID- 9724843 TI - Seizures in the de-institutionalized patients with early brain injuries. AB - The number of individuals with early brain injury and multiple disabilities discharged from institutions had increased steadily over the past 10 years. The most frequently encountered problems by patients with early brain injury and epilepsy placed in the community were reviewed. Unavailability of specialized services, inadequate reimbursement, high turnover and lack of knowledge by caregivers, lack of understanding/acceptance of the concepts of seizure intractability and pseudoseizures, unrealistic expectations by overseeing agencies and/or family, and side effects of antiepileptic medications were the most common seizure-related problems afflicting these patients. Everything else being equal, good exchange of information between the various caregivers and continued education of caregivers, both time consuming endeavors, are the crucial factors for the successful management of seizures in this population. PMID- 9724842 TI - Cases of two patients whose food aversions disappeared following severe traumatic brain injury. AB - Following traumatic brain injury (TBI) the complete disappearance of food aversions was observed in the cases of two patients. In one of the cases, a young female, this change in her food aversion was manifested several months after the TBI, from the time when she was able to eat normally. The other patient, a young man, exhibited the disappearance of his food aversion immediately after recovery from his unconscious state following TBI. These results indicate that the disappearance of food aversions was a consequence of TBI. PMID- 9724844 TI - Paroxetine and amotivational syndrome. PMID- 9724845 TI - From naive to memory. Development and regulation of CD4+ T cell responses. AB - Specific immune responses proceed through and are regulated at several stages: activation of naive cells and their differentiation into effector cells, completion of effector functions, development of memory cells, and subsequent reactivation of memory cells. To understand the development and regulation of CD4+ T cells in immune responses, naive CD4+ T cells were enriched from T cell receptor (TCR) transgenic mice, and used to generate effector and memory populations in vivo and in vitro. The expression of a common TCR on all of these developmental subsets has allowed us to compare directly their phenotype, cytokine profiles, activation requirements, and susceptibility to apoptosis. Our experiments have revealed interesting distinctions among naive, effector, and memory subsets of CD4+ T cells and have important implications for our understanding of immune responses. PMID- 9724846 TI - A gene therapy approach to treatment of autoimmune disease. AB - New insights into the underlying mechanisms for the development of autoimmune diseases in humans and various animal models continue to increase with our understanding of factors that drive polarization of T helper (Th) responses and tolerance. This information has led to the development of new treatment strategies, including oral tolerance clinical trails and the use of altered peptide ligands in animal models. These approaches have shown some promise and provided additional insight into the disease processes. The use of gene therapy in many disease states continues to increase. We are starting to see the application of gene therapy in chronic diseases in humans. Gene therapy has been used in several animal models of autoimmune disease with promising preliminary results. In this article, an overview will be provided for the use of gene therapy in autoimmune disease. PMID- 9724847 TI - Immune cell signaling aberrations in human lupus. AB - A large array of heterogeneous aberrations of the immune system have been described in systemic lupus erythematosus (SLE). Since the function and the fate of the immune system cells are governed principally by the biochemical events that follow ligation of specialized cell-surface receptors, we will review in this article recent developments in our understanding of abnormalities in the biochemistry of signals generated either by the antigen-receptor complex or by systems of costimulatory cell-surface molecules, like the CD28/CTLA4:CD80/CD86 and the CD40:CD40L pairs found on the surface membrane of lupus immune cells. PMID- 9724848 TI - Intestinal intraepithelial T lymphocytes. Our T cell horizons are expanding. AB - The alimentary tract is an essential structure for the ingesting of nutrients from the outside, and even most primitive animals have a straight tract that runs from the mouth to the anus. We come into contact with the outside world through our skin and mucous membranes. The surface area of the enteric mucous membrane, which absorbs nutrients, is enlarge through its ciliary structure, and the enteric cavity creates by far the largest external world that we come into contact with. For instance, the enteric mucosal surface of the human gastrointestinal tract covered by a single layer of epithelial cells corresponds to the size of one-and-a-half tennis courts, and the innumerable number of epithelial cells covering this mucous surface are entirely replaced by new epithelial cells in the space of just several days. Simultaneously, the fact that 60-70% of peripheral lymphocytes are congregating in the gastrointestinal tract supports the notion that the enteric mucous membrane represents an extremely dangerous locale, where numerous harmless/precarious external antigens come in through the wide array of food we injest on a daily basis, and the literally infinite amounts of normal intestinal flora intermingled from time to time with life-threatening microbes surge across. Surprisingly, approximately one out of the five cells in the intestinal epithelium are lymphocytes, most of which are ill-defined T cells having unusual, but distinctive characteristics and situated apparently so close to external antigens in the entire body. This article deals with the information that has been accumulated mainly in the past decade concerning the development, phenotypes, and possible function of these yet unacknowledged mucosal T cells that lurk in the anatomical front of the intestine. PMID- 9724849 TI - Partial lipodystrophy, mesangiocapillary glomerulonephritis, and complement dysregulation. An autoimmune phenomenon. AB - Partial lypodistrophy (PLD) is a rare disease in which, there is loss of fat usually from the upper part of the body. The disease is frequently associated with mesangiocapillary (membranoproliferative) glomerulonephritis Type II (MCGN II). In the early 1970s, it was noticed that MCGN II and/or PLD was sometimes associated with dysfunction of the complement system as reported in several case descriptions and studies. Subsequently, an IgG autoantibody was detected-C3 nephritic factor (C3NeF). The target of this autoantibody is the alternative pathway C3 convertase-C3bBb. There are sporadic case reports that linked PLD, MCGNII, and C3NeF with autoimmune diseases. This association may be more than a coincidence. The complement deficiency may lead to perturbation of the immune system, which may trigger some of the autoimmune diseases. This article will be focused on the association among PLD, MCGN II and C3NeF. PMID- 9724850 TI - A sequential study of circulating immune complexes, complement and immunoglobulins in borderline tuberculoid leprosy patients with and without reactions. AB - Sequential estimates of the levels of circulating immune complexes (CIC), complement catabolic fragment C3d, complement-mediated immune complex solubilization (CMS) and immunoglobulins were made in 24 newly diagnosed with borderline tuberculoid leprosy over a 20 month period after initiation of chemotherapy. Fourteen of these patients had not suffered from reversal reactions either at the time of presentation or during the follow-up. The levels of CIC were evaluated in them from the third to the eleventh month after starting chemotherapy and immunoglobulin G (IgG) levels were evaluated up to eight months. The concentrations of C3d and immunoglobulins A (IgA) and M (IgM) were normal in these patients. The other ten patients had reversal reaction at the time of diagnosis which subsided by the third month after starting treatment. They did not have reversal reactions later. The levels of CIC and IgG were elevated and those of CMS were depressed throughout the study period. Serum C3d level was initially elevated but came down to normal by the third month while IgA and IgM levels were within normal limits. The relevance of these findings to the genesis of reversal reaction is discussed in this communication. PMID- 9724851 TI - Is PGL-1 also present in Leishmania donovani promastigotes? AB - A soluble antigen complex (SAC) derived from the ruptured promastigotes of Leishmania donovani parasites (LD-SAC) was used for complement fixation test (CFT) in leprosy Cases of tuberculoid and borderline tuberculoid leprosy, post kala azar dermal leishmaniasis (TT, BT, PKDL) and control sera gave negative CFT. Smear-positive cases of borderline (BB, BL) and lepromatous (LL) leprosy and drug resisting cases of pulmonary tuberculosis gave positive CFT; smear-negative cases of LL leprosy sera also gave positive CFT. Sera of smear-negative inactive LL patients contained only PGL-1 and PDIM antigens for a long time after they become inactive. Therefore, the positive CFT in inactive LL makes us suspect whether PGL 1 is present in LD promastigotes. PMID- 9724852 TI - Studies on rapid assessment methods in leprosy. AB - A study was undertaken in Pudukottai district, Tamilnadu, India to test rapid assessment methods: viz (i) sample surveys with lower coverages for clinical examination in estimating the disease problem in the community, (ii) utility of registered case prevalence for estimating the actual prevalence in a given area, (iii) leprosy in school-going children and its utility in estimating leprosy prevalence in the community, and (iv) information on disability and smear positivity in estimating leprosy prevalence; and develop correction factors for estimating leprosy situation. A sample of 23 clusters from 582 clusters of contiguous villages and hamlets was further divided into two random sub-samples for two surveys with differing coverages. One team covered nine clusters comprising 34 villages with a population of 17,562 and examined 15,596 with a population of 26,927 and examined 16,622 (62%) persons for leprosy. The results showed that: (i) leprosy sample surveys with lowered coverages would tend to miss valuable information, in terms of quality and quantity; (ii) from 'known case' registers, to estimate the true burden of leprosy disease and to monitor its trend over time is inadequate; (iii) school surveys are of limited value for estimating the disease burden in the community or to monitor its trend over time; (iv) the number of smear-positive cases is to small to serve as an indicator for the total case load in the community; and (v) the prevalence of active disease and that of grade 2 disability in the community are poorly correlated. Reliable methods other than those used here need to be developed for evaluation and monitoring of the disease burden particularly in the post-MDT era. PMID- 9724853 TI - Neoplastic transformation of chronic ulcers in leprosy patients--a retrospective study of 23 consecutive cases. AB - A retrospective analysis of chronic ulcers among leprosy patients seen over the last 20 years yielded 23 cases of neoplastic transformation. It showed a peak at the sixth decade, an incidence of 3.66/100 among hospitalised ulcer cases and male/female ratio of 1.6:1. Borderline tuberculoid was the most common type of leprosy involved (40%). Squamous cell carcinoma was the most common neoplasia. Its usual site was plantar ulcers. Heel ulcers showed relatively greater predeliction for malignancy (38.5%). Histopathological proof of malignancy is desirable and that may require multiple biopsies. Metastasis is rare but potentially fatal. The surgical treatment must provide a functional, trouble-free limb. Forefoot or Lisfranc's amputation for distal third ulcers and below-knee amputation for large midfoot and ulcers are procedures of choice. Wide excision may be used in select cases. PMID- 9724854 TI - Clinical ocular study in leprosy patients at a sanitary dermatological hospital in Brazil. AB - This study is based upon the observation f 363 leprosy patients having different types of the disease. At the time of the examination, we did not have any previous knowledge about the type of leprosy the patients were having. Thus, the eye examination was done without the knowledge of clinical diagnosis. The ocular examination protocol covered the following: visual activity, facial muscle function, eyebrows, eyelashes, lacrimal system, pupil, eye motility, corneal sensitivity, Schirmir's test and study of the anterior segment of the ocular bulb with a slit-lamp. The study patients included 275 cases of lepromatous leprosy, 57 tuberculoid, 29 indeterminate and two dimorphous cases. The age of the patients ranged between 18 and 82 years, and 229 of them were males. Among those patients, 183 were whites, 23 were black and 157 were mulatto. PMID- 9724855 TI - Study of ocular changes in leprosy patients. AB - In this study, 997 leprosy patients were examined, 528 of them with lepromatous leprosy (53%), 199 with borderline leprosy (20%), 167 with tuberculoid leprosy (16%) and 103 (10.3%) with indeterminate leprosy. Changes in the ocular bulb were noted in 314 patients (31.5%) specially in those with lepromatous leprosy. These alterations were greater with increasing age of the patient and length of disease. Severe ocular lesions were rare, probably due to previous systemic treatment. The "pearls" in the fundus of the eye resulting from leprosy were also studied. PMID- 9724856 TI - Utilizing primary health care workers for case detection. AB - Under the National Leprosy Elimination Programme it takes at least one year for the paramedical worker to survey the allotted population for case detection. An alternative strategy in warranted for States like Bihar still having a high case load and poorly functioning leprosy programme. An intensive case finding programme using Primary Health Care (PHC) workers was organized in Bhojpur district, Bihar State, India. The whole population (3, 173, 701 in 1996) of the district was screened within a period of four days and confirmation of suspected cases was carried out in four days. During this screening procedure, 1586 new leprosy cases were detected (NCDR = 5 cases per 10,000) and all were started on MDT. The new cases constituted 26.4% of active cases existing on record before the screening. After this experience, the prevalence rate of active cases increased from 19 to 24 10,000. If such rapid screening programmes are done at least twice a year, it will greatly hasten the process of elimination of leprosy. PMID- 9724857 TI - Polyarteritis nodosa masquerading as neuritic leprosy. PMID- 9724858 TI - Single dose multidrug therapy for single lesion paucibacillary leprosy. PMID- 9724859 TI - Single dose multidrug therapy for single lesion paucibacillary leprosy. PMID- 9724860 TI - Single dose multidrug therapy for single lesion paucibacillary leprosy. PMID- 9724861 TI - Single dose multidrug therapy for single lesion paucibacillry leprosy. PMID- 9724862 TI - Shortening duration of treatment of multibacillary leprosy. PMID- 9724863 TI - Hidden cases of leprosy. PMID- 9724864 TI - Delayed resolution versus treatment failure in PB leprosy patients. PMID- 9724865 TI - Bacteriological status of newly registered cases. PMID- 9724866 TI - Primary amyloidosis: not a cause of peripheral nerve enlargement. PMID- 9724867 TI - Dapsone syndrome: an incomplete form. PMID- 9724868 TI - Joint ILEP/WHO workshop on reaching undetected leprosy patients in endemic countries, Geneva, Switzerland, 18-19 July 1997. PMID- 9724869 TI - Report of an informal consultation on "Community action for the elimination of leprosy" Dhaka Bangladesh 5-6 March 1997. PMID- 9724870 TI - Apical organelles and host-cell invasion by Apicomplexa. AB - Host-cell invasion by apicomplexan parasites involves the successive exocytosis of three different secretory organelles; namely micronemes, rhoptries and dense granules. The findings of recent studies have extended the structural homologies of each set of organelles between most members of the phylum and suggest shared functions of each set. Micronemes are apparently used for host-cell recognition, binding, and possibly motility; rhoptries for parasitophorous vacuole formation; and dense granules for remodeling the vacuole into a metabolically active compartment. In addition, gene cloning and sequencing have demonstrated conserved domains, which are likely to serve similar functions in the invasion process. This is especially true for microneme proteins containing thrombospondin-like domains, which are likely to be involved in binding to sulphated glycoconjugates. One such protein was recently shown to be required for the motility of Plasmodium sporozoites. These molecules have been shown to be shed on the parasite and/or cell surfaces during the invasion process in Plasmodium, Toxoplasma and Eimeria. For rhoptries and dense granules, the association between exocytosed proteins and the parasitophorous vacuole membrane had been analyzed extensively in Toxoplasma, as these proteins are likely to play a crucial role in metabolic interactions between the parasites and their host cells. The development of parasite transformation by gene transfection has provided powerful tools to analyze the fate and function(s) of the corresponding proteins. PMID- 9724871 TI - Structure and function of the parasitophorous vacuole in Eimeria species. AB - The intracellular life-cycle of Eimeria are located in the host cell within a membrane-bound parasitophorous vacuole. The invasion process and the formation of the parasitophorous vacuole are mediated by characteristic organelles within the apical complex. During invasion, the parasitophorous-vacuole membrane is manipulated by the parasite and functions later in the development cycle as a molecular sieve, allowing the exchange of metabolites between parasite and host cell. Unlike the cyst-forming coccidia, there is little evidence of parasitophorous-vacuole membrane transformation in the later stages of the lifecycle of Eimeria species. Compared with the human pathogens Plasmodium and Toxoplasma, rather little is known about the parasitophorous vacuole and parasitophorous-vacuole membrane of animal pathogens of the genus Eimeria. PMID- 9724872 TI - Advances in the life cycle of Toxoplasma gondii. AB - This paper reviews recent studies on the life cycle of Toxoplasma gondii. Tachyzoites, bradyzoites, and sporozoites are the three infectious stages of T. gondii. Humans and animals become infected mainly by ingesting bradyzoites or oocytes. After ingestion, both bradyzoites and sporozoites convert to tachyzoites inside tissues. The conversion of tachyzoites to bradyzoites and bradyzoites to tachyzoites is of biological and clinical significance because bradyzoites are less susceptible to chemotherapy and reactivation of bradyzoites to tachyzoites is considered the cause of fatal toxoplasmosis in AIDS patients. Of all the methods currently available to assess stage conversion of T. gondii, feeding infective stages to cats is the most reliable method. Felidae, the definitive hosts of T. gondii excrete oocysts 3-10 days after ingesting tissue cysts/bradyzoites, > or = 18 days after ingesting oocysts, and > or = 13 days after ingesting tachyzoites. PMID- 9724873 TI - Toxoplasma gondii: ESTs and gene discovery. AB - Partial cDNA sequences or Expressed Sequence Tags (ESTs) have proven to be an economical way to gain information about expressed genes in a variety of organisms. Further, ESTs can be generated for strain or developmental stage comparisons. Currently there are over 10, 000 ESTs for Toxoplasma gondii derived from RH tachyzoite, ME49 tachyzoite and ME 49 bradyzoite cDNA libraries. A set of Web pages and tools have been developed to proved easy access and rapid analysis of these data. Top Hits lists, T. gondii-specific databases/search tools and cluster analyses can be browsed or used to rapidly gain insight into the structure and potential function of genes/proteins held within the database. The previously characterized Eimeria protein Etp 100 has been used to demonstrate how it is possible to use these tools to extract and assemble information about the putative T. gondii homologue. PMID- 9724874 TI - A polymorphism in a DNA polymerase alpha gene intron differentiates between murine virulent and avirulent strains of Toxoplasma gondii. AB - The IC intron, found within the DNA polymerase alpha gene of Toxoplasma gondii, was used to evaluate the genetic relationship among 10 strains of T. gondii. Sequence comparison detected polymorphisms within this 652 bp intron which correlated with murine virulence. The results reported here suggest that T. gondii contains two lineages, corresponding with their virulence, evolving independently following their separation. The extensive homology of the IC sequences within the virulent and avirulent groups affirms the close relationship of the strains within the group, as reflected by the identical nucleotide substitutions and dinucleotide insertions/deletions observed. In addition, the presence of the Nde I restriction enzyme site within the IC intron of avirulent strains allows definition of a T. gondii strain as murine virulent or avirulent without needing to test it in vivo. PMID- 9724875 TI - Gene sequence and transcription differences in 70 kDa heat shock protein correlate with murine virulence of Toxoplasma gondii. AB - This study compared the genes encoding cytoplasmic 70 kDa heat shock protein in virulent and avirulent strains of Toxoplasma gondii, to determine whether differences may contribute to the variation in protein expression levels previously reported for this protozoan parasite. A T. gondii PCR probe with homology to Eimeria acervulina cytoplasmic 70 kDa heat shock protein was used to screen a genomic DNA mini-library and isolate the gene from the virulent RH strain. The entire coding region was subsequently amplified from the avirulent ME49 strain by PCR. Alignment of the gene sequences revealed that the virulent RH strain had four copies of a seven-aa repeat unit (GGMPGGM) at the 3'- end of the gene compared with five copies in the avirulent ME49 strain. Comparison of this region among other virulent and avirulent strains revealed that this difference was consistent with virulence. Copy number estimation revealed that this gene is single-copy in both the RH and the ME49 strains. Analysis of mRNA expression revealed a 1.5- to 2-fold increase in transcription of this gene in virulent strains when compared with avirulent strains. For each strain, mRNA was observed at similar levels whether grown in vivo or in vitro. Also, heat-shock treatment of tachyzoites prior to harvest did increase mRNA levels in vitro. This suggests that post-transcriptional regulation of cytoplasmic 70 kDa heat shock protein may occur in T. gondii. Differences in cytoplasmic 70 kDa heat shock protein have been demonstrated at genomic and transcriptional levels in virulent strains compared with avirulent strains, suggesting that this 70 kDa heat shock protein may play an important role in the virulence of T. gondii. PMID- 9724876 TI - Polymerase chain reaction approaches for the detection of Neospora caninum and Toxoplasma gondii. AB - This review summarizes existing knowledge on the development and use of the polymerase chain reaction for the detection of DNA from Neospora and Toxoplasma. Several strategies which utilise the polymerase chain reaction for the diagnosis of toxoplasmosis in humans and livestock have been described and they principally target the B1 repetitive sequence, the P30 gene or ribosomal DNA. Experience has shown that the polymerase chain reaction has proven insufficiently robust to serve as a diagnostic test alone although when used in conjunction with other diagnostic techniques it does prove to be a useful aid. The marketing of a commercial polymerase chain reaction kit may well solve some of the inadequacies seen using "home made" polymerase chain reaction technology which are commonly used in diagnostic laboratories around the world. Recent progress on the development of polymerase chain reaction diagnostics for Neospora has been rapid and is discussed in detail. PMID- 9724877 TI - Genetic analysis of the essential components of the immunoprotective response to infection with Eimeria vermiformis. AB - The immune responses generated after infection with Eimeria spp. are complex, include both cellular and humoral components, and lead to protection against re infection. To facilitate the rational development of the next generation of anticoccidial vaccines it is important that the nature of the immunoprotective response against infection with Eimeria spp. is determined. In this brief report we discuss results that were obtained using a combination of genetic and cellular approaches to dissect the essential immune effector components that operate against infection with Eimeria vermiformis. Mice rendered deficient of immune function by targeted gene disruption at a variety of immune loci represent an integral component of our studies and include those with targeted gene disruption at loci that encode the B- and T-cell receptors (BCR, TCR), antigen presentation molecules and immune-effector molecules. Our studies demonstrated that TCR-alpha beta + T cells are essential for immunoprotection during both primary and secondary infection. Moreover, during primary infection the major effector cell type is a population of major histocompatibility complex class II-restricted, interferon-gamma-producing TCR-alpha-beta T cell consistent with a T helper 1 phenotype. In addition, there is a supplementary role for another class of cells (presumably T cells) that are restricted to either non-classical antigen presentation molecules or classical major histocompatibilty complex class I loaded via an atypical pathway. Mice with a deficiency in interleukin-6 were slightly more susceptible to primary infection than intact animals, consistent with the reported effects of interleukin-6 upon the generation of T helper 1-type responses in vivo. In terms of the host response to re-infection, TCR-alpha-beta T cells were essential for immunity, but the requirement for specific cell subsets and effector mechanisms was much less stringent. Mice deficient in gamma delta T cells, classical major histocompatibility complex class I, non-classical antigen presentation pathways, the cytokines interferon-gamma, interleukin-4, interleukin-6 and the cytolytic effector molecules perforin or FasL were completely immune to secondary infection. Moreover, major histocompatibility complex class II-deficient I-A-beta-/- mice were capable of mounting a substantial response to secondary infection, manifest by a 95% reduction in oocyst output compared with primary infection. These data have important consequences for the development of immune intervention strategies and indicate that vaccine development may be targeted toward the generation of a wider range of effector mechanisms than those that operate during primary infection. PMID- 9724878 TI - Role of T lymphocytes and cytokines in coccidiosis. AB - Development of a vaccine for avian coccidiosis has been hampered by lack of understanding of the various components of the host immune system leading to protective immunity. Clear understanding of the cellular dichotomy in cytokine production in mice and the availability of immunological reagents, as well as gene knock-out mice, now makes in-depth immunological study in this species feasible. From studies of various parasitic infection models in mice, it is becoming clear that complex regulation by cytokines is involved in host immunity. Furthermore, the studies in mice clearly indicated an important role of various effector mechanisms involving T lymphocytes, macrophages, natural killer (NK) cells and cytokines in resistance to coccidiosis. In comparative studies of coccidiosis in chickens, in-vivo and in-vitro studies revealed that interferon gamma, tumor necrosis factor and transforming growth factor-beta are induced following Eimeria infection. Depletion studies revealed the importance of CD8+TCR alpha-beta+ T lymphocytes in host protective immunity to avian coccidiosis. Taken together, studies in mice and chickens are providing a better understanding of the role of effector cells and soluble factors which control immune responses to Eimeria parasites. PMID- 9724879 TI - Bovine T cell responses to Cryptosporidium parvum infection. AB - To better understand the immune mechanisms important for clearing of the primary infection and the subsequent development of resistance to Cryptosporidium parvum infection, several groups have recently characterised changes within the lymphoid cell population of the intestinal mucosa and associated lymphoid tissue in calves with cryptosporidiosis. In naive animals, infection results in a significant increase in the number of CD4+ and CD8+ T cells present within the intraepithelial lymphocyte population, lamina propria and Peyer's patch of the ileum. This is accompanied by a rapid and transient increase in the number of gamma/delta T cells present within the intestinal villi. In response to a challenge infection in immune calves, there is a substantial increase in the number of CD4+ T cells present in the Peyer's patch of the ileum and a specific localization of CD8+ T cells to the epithelium of the intestinal villi. Together, these data demonstrate that C, parvum elicits a strong cell-mediated response following both primary and secondary infections in calves, and that CD8+ T cells may play an important role in the bovine immune response to C. parvum infection. PMID- 9724880 TI - Epidemiological aspects of the use of live anticoccidial vaccines for chickens. AB - This review address the epidemiology (epizootiology) of coccidiosis in commercial chickens with emphasis on the effects on the use of live vaccines. Surveys suggest that all seven valid species of chicken coccidia (Eimeria acervulina, Eimeria brunetti, Eimeria maxima, Eimeria mitis, Eimeria necatrix, Eimeria praecox and Eimeria tenella) are ubiquitous. All species are pathogenic to various extents. New results are presented on the pathogeneicities of E. acervulina, E. mitis and E. Praecox. Unless ingested by chickens, oocysts in poultry-house litter may die after about 3 weeks. Oocyst sporulation may be better in drier, rather than wetter, litter. Whether sporulated or not, up to 20% of ingested oocysts may pass undamaged through a chicken's intestine. The excreted, sporulated oocysts can be immediately reingested to initiate an infection; the unsporulated oocysts can still sporulate after passing through the intestine. The seven species differ in their times of appearance in commercial flock; hence particular vaccines may be designed for rearing standard broilers for up to about 6 weeks or for breeding stock. Attenuated, precocious lines of Eimeria in vaccines have low reproductive potentials, thus avoiding crowding, developing optimally, and stimulating immune response with minimal tissue damage. Cross-immunity between Eimeria species is probably minimal. There is reciprocity between the immune status of chicken and their excretion of oocysts for each species, ensuring continual stimulation of immune responses in birds on litter. Paracox vaccine, comprising all seven Eimeria species, is shown here to stimulate immunity to each of them independently. Total oocyst accumulation in litter following Paracox vaccination at 1 week comprises a small peak of vaccinal oocysts at 2-4 weeks, then a higher peak of the local virulent population at 4-7 weeks, which rapidly wanes. The attenuated drug-sensitive vaccinal oocysts probably interbreed with the corresponding wild species, reducing both virulence and drug-resistance in the local population. Anticoccidial vaccines may not induce complete immunity in chickens with lowered immunocompetence due to stressors, including certain viral disease. Future development of live vaccines for standard broilers may be expected in the relatively short term. The useful lives of anticoccidial drugs might be extended by rotating them with live vaccines. PMID- 9724881 TI - Use of live oocyst vaccines in the control of avian coccidiosis: experimental studies and field trials. AB - Areas addressed in this study on the use of live oocyst vaccines to control coccidiosis include: the influence of immunocompetency of the strains and sex of the birds used; methods of delivery of vaccine; immunological variation between different strains of the same coccidial species; and the effects of combining vaccine with anticoccidial medication. The results show that vaccination with live oocysts elicited significant protection against coccidiosis, both with experimentally induced and naturally acquired coccidial infection, resulting in average bird weight gains and feed efficiency similar to that obtained with conventional anticoccidial medication. PMID- 9724882 TI - Progress on developing a recombinant coccidiosis vaccine. AB - The past 10 years of research aimed at developing subunit vaccines against a number of apicomplexans, including Eimeria, Plasmodium and Toxoplasma, have, if anything, revealed the complex nature of parasite-host interactions. The Knowledge gained from this research has shown why developing a subunit vaccine based on a single recombinant antigen from one developmental stage of the parasite was an overly optimistic approach. Many apicomplexan parasites have acquired unique strategies to evade host immunity. The variable expression of genes encoding erythrocyte membrane protein 1 of Plasmodium falciparum [1] (Berendt et al. Parasitology 1994;108:S19-S28) exemplifies one such strategy. The particular mechanism for evading immune destruction depends on a number of interrelated factors, not least of which is the parasite life-cycle and the availability of susceptible hosts. The goal of any vaccine, be it an attenuated organism or a recombinant antigen, is to break the cycle of infection. The development of a recombinant vaccine against apicomplexan parasites will depend on identifying those antigens and intracellular processes that are vital to the parasite survival and those which exist merely as a way of evading immunity. The information that follows is a review of both molecular biology/biochemistry of eimerian parasites and factors that influence host immune responses to coccidia. PMID- 9724883 TI - Progress in recombinant vaccine development against coccidiosis. A review and prospects into the next millennium. AB - The increasing problems encountered by the poultry industry, despite the extensive use of drugs, have emphasised the need for an immunological solution for the economic damage caused by the Eimeria parasite. Although immunity develops relatively fast following a natural infection, to induce protection by using parasite extracts or single antigens appears more difficult. Nevertheless, the development of a vaccine based on defined antigens seems the best solution in the long run. At the VIth International Coccidiosis Conference in 1993 the first promising results were reported from small-scale experiments using recombinant antigens. This review summarises the advances in this field of research from 1993 onwards. Although since then not many reports have been published about the effects of using recombinant. antigens as a vaccine against coccidiosis, a number of interesting new proteins which could be considered good targets for such a vaccine have been described and are referred to herein. Proteins involved in the process of invasion of the host cell by the extracellular parasite are regarded as key components in the developmental cycle of the parasite. These components possibly bind to receptors on the host cell. Interference with this process could be a target of the protective immune response. Progress has also been made in characterising the immune mechanisms activated by infection with the parasite. From experimental mouse models and from studies in chickens, a better insight has been obtained towards the involvement of CD4- and CD8-type T cells in, respectively, the inductor and the effector branch of the immune response, although not all questions have been answered. Several antigens have been selected using T-cell stimulation and cytokine assays and these are reviewed. In a third section, mostly unpublished results of our own experiments dealing with the use of live vectors to present defined antigens such as Ea1A and EaSC2, a parasite refractile body transhydrogenase and a lactate dehydrogenase, respectively, are summarised. Partial protection could be induced using Salmonella typhimurium as a carrier for these antigens, in that the oocyst output was reduced by up to 50% after challenge and weight gain could be improved by 5 10% over non-vaccinated challenged chickens, when tested in a floor-pen trial. Similar results were obtained when these antigens were presented by viral vectors such as Fowlpox virus or Herpes virus of turkey. These data seem to offer good prospects for the accomplishment of a safe and efficaceous vaccine based on recombinant DNA technology. These expectations are corroborated by recent breakthrough in transfection of related parasites such as Plasmodium and Toxoplasma gondii, and by the increasing amount of genomic information becoming available every day, the impact of which cannot even be estimated yet. These new technologies will allow us to solve the complex problems that we once created ourselves. PMID- 9724884 TI - Dietary modulation of avian coccidiosis. AB - During the past several years, our laboratory has been investigating the anticoccidial activities of various natural products that have potential use as dietary supplements for coccidiosis control. Sources of fats containing high concentrations of n-3 fatty acids such as menhaden oil and flaxseed oil and flaxseed, when added to starter rations and fed to chicks from one day of age, effectively reduce lesions caused by the caecal parasite Eimeria tenella, but not lesions caused by Eimeria maxima. Our results are consistent with reports of effects of diets high in n-3 fatty acids on other protozoan parasites which suggest that the state of oxidative stress induced by these diets in the cells of both host and parasites is responsible for their parasitic actions. Artemisinin, a naturally occurring (Artemisia annua) endoperoxide and effective antimalarial significantly lowers lesions from E. tenella when given at low levels as a feed additive. The mechanism of its action is also considered to involve induction of oxidative stress. Diets supplemented with 8 p.p.m. gamma-tocopherol (abundant in flaxseeds) or with 1% of the spice tumeric, reduce mid-small intestinal lesion scores and improve weight gains during E. maxima infections. These compounds may exert their anticoccidial activity because they are effective antioxidants. Betaine, a choline analogue found in high concentrations in sugar beets, improves nutrient utilisation by animals under stress. When provided as a dietary supplement at a level of 0.15% it has enhanced the anticoccidial activity of the ionophore, salinomycin. Betaine may act as an osmoprotectant whereby it improves the integrity and function of the infected intestinal mucosa. In in vivo studies, betaine plus salinomycin significantly inhibit invasion of both E. tenella and E. acervulina. However, subsequent development of E. acervulina is inhibited more effectively with this combination treatment than development of E. tenella. PMID- 9724887 TI - Commentary: compasses to help navigate the terrain on mental health classification. AB - OBJECTIVE: To study current patterns of panic disorder (PD) recognition and management by primary care physicians (PCPs). METHOD: We administered a vignette describing a female PD patient to 189 PCPs. RESULTS: Three-quarters of respondents believed that PD was at least 50 percent probable, and the mean PD likelihood rating was 63 percent. Diagnostic suspicion was significantly higher for PD than for other anxiety disorders, major depressive disorder, and cardiac disorders. Medication was rated as significantly more necessary than medical testing and mental health referral. A benzodiazepine was suggested by 78 percent of respondents, while 35 percent suggested a serotonin reuptake inhibitor (SRI). Under half rated the patient as requiring medical testing, mostly for hyperthyroidism (70%) and/or cardiovascular disorder (62%), and half felt that the patient required mental health referral. CONCLUSIONS: The data suggest that most PCPs are able to recognize PD. However, they may be excessively inclined to prescribe benzodiazepines rather than more appropriate medications. PMID- 9724885 TI - Evaluation of the efficacy of anticoccidial drugs against Eimeria species in the fowl. AB - Anticoccidial drugs undergo a vigorous series of test during development in order to demonstrate their efficacy and safety. Evaluation of efficacy may also be necessary after a drug is utilised in the field in order to establish whether resistance has developed. The principles and procedures used to evaluate anticoccidial efficacy were reviewed in a series of papers presented at a symposium held at the University of Georgia in May 1969. It is a testimony to the quality of this symposium that much of the information presented remains valid today. In the USA, general procedures that should be followed in new drug investigations are given in various guidelines and memoranda issued by the Food and Drug Administration. Specific guidelines relate to anticoccidial investigations. Although not a legal requirement, they provide a standard for those investigators seeking approval of a new anticoccidial drug. These guidelines are periodically revised, and investigators should consult the FDA for advice on specific protocols. Many of these procedures are also appropriate for the determination of drug resistance. PMID- 9724886 TI - The classification of mental disorders in primary care: a guide through a difficult terrain. AB - BACKGROUND: Primary care physicians traditionally have a strong interest in the mental health of their patients. Three classification systems are available for them to diagnose, label, and classify mental disorders: 1) The ICD-10 approach with three options, 2) The DSM-IV approach with two options, and 3) the ICPC approach with two options. This article lists important similarities and differences between the systems to help potential users choose the option that best meets their needs. METHODS: Definitions for depressive disorder, anxiety disorder, and somatization disorder are compared on five characteristics of classification: 1. the domain, 2. the scope, 3. the nature of the definitions, 4. focus on episodes of care, and 5. clinical guidelines. RESULTS: Primary care physicians and psychiatrists have different perspectives, reflected in different classifications. Each system has specific possibilities and limitations with regard to the diagnosis of mental disorders. For common mental disorders it is possible, however, to choose codes from one system while maintaining compatibility with the other two. Comparability as to the diagnostic content of the different classes, however, is more difficult to establish. The available classification systems give both primary care physicians and psychiatrists options to diagnose, label, and to classify mental disorders from their own perspective, but once a system has been chosen the clinical comparability of a patient with the same diagnosis in other systems is limited. CONCLUSION: Compatibility among systems can be optimized by strictly following a number of rules. The conversion between ICPC and ICD-10 (and consequently DSM-IV) allows simultaneous use of ICPC and ICD-10 as a classification and DSM-IV as the standard nomenclature. This is of particular interest for computer based patient records in primary care. The clinical comparability of the same diagnosis in different systems however is limited by the characteristics of the different system. PMID- 9724888 TI - Primary care physician responses to a panic disorder vignette: diagnostic suspicion and clinical management. PMID- 9724889 TI - The relationship between religious activities and blood pressure in older adults. AB - OBJECTIVE: To examine the relationship between religious activities and blood pressure in community-dwelling older adults. METHOD: Blood pressure and religious activities were assessed in a probability sample of 3,963 persons age sixty-five years or older participating in the Duke EPESE survey. Participants were asked if their doctor had ever informed them that they had high blood pressure and if they were currently taking medication for high blood pressure. After the interview, systolic and diastolic blood pressure were measured following a standardized protocol. Data were available for three waves of the survey (1986, 1989-90, and 1993-94). Analyses were stratified by age (65-74 vs. over 75) and by race (Whites vs. Blacks) and were controlled for age, race, gender, education, physical functioning, body mass index, and, in longitudinal analyses, blood pressure from the previous wave. RESULTS: Cross-sectional analyses revealed small (1-4 mm Hg) but consistent differences in measured systolic and diastolic blood pressures between frequent (once/wk) and infrequent (< once/wk) religious service attenders. Lower blood pressures were also observed among those who frequently prayed or studied the Bible (daily or more often). Blood pressure differences were particularly notable in Black and younger elderly, in whom religious activity at one wave predicted blood pressures three years later. Among participants who both attended religious services and prayed or studied the Bible frequently, the likelihood of having a diastolic blood pressure of 90 mm Hg or higher was 40 percent lower than found in participants who attended religious services infrequently and prayed or studied the Bible infrequently (OR 0.60, 95% CI, 0.48-0.75, p < .0001). Among participants told they had high blood pressure, religiously active persons were more likely to be taking their blood pressure medication; this could not, however, explain the differences in blood pressure observed. While most religious activity was associated with lower blood pressure, those who frequently watched religious TV or listened to religious radio actually had higher blood pressures. CONCLUSIONS: Religiously active older adults tend to have lower blood pressures than those who are less active. This applies to attendance at religious services and private religious activities, but not to religious media. Physiological mechanisms are discussed. PMID- 9724891 TI - Identification and classification of factitious disorders: an analysis of cases reported during a ten year period. AB - OBJECTIVE: The current article offers a new conceptualization of factitious disorders based on cases reported in the literature. METHOD: The current analysis examines twenty-nine cases of factitious disorder patients over the course of ten years (1986-1996). Cases were found through PSYCHLIT and MEDLINE searches. Patient variables examined include: demographics, occupational status, marital status, childhood history, extent of medical history, the presence of a borderline personality disorder, and presence of a psychosocial stressor prior to the onset of the factitious disorder. RESULTS: An examination of the cases found demographic patterns of illness presentation consistent with previous reviews of the disorder. From the examination emerged two distinct types of factitious presentations-one acute, one chronic. A two-dimensional approach is introduced in an attempt to understand various disease presentations. Cases were classified based on proposed Current Life Stress and Chronic Life Pattern dimensions. These dimensions are measures of the extent to which the patient's factitious presentation is in response to an immediate psychosocial stressor, or an action consistent with a long-term maladaptive behavior pattern resulting from an underlying character pathology. Three patient groups were identified based on estimated patient levels of each dimension. The three groups are: Stress Response, Life Response, and Mixed Response. CONCLUSION: Recommendations are made to increase the role of physicians in the detection of factitious patients, as well as to move toward a more uniform reporting of cases of factitious disorders in the literature. PMID- 9724890 TI - Use of the Brief Psychiatric Rating Scale as a predictor of psychiatric admission for non-suicidal patients. AB - OBJECTIVE: We hypothesized that a systematic determination of symptom severity would predict psychiatric admission for non-suicidal patients referred for a psychiatric evaluation in an urban emergency department (ED) setting. METHOD: In a pilot study involving consecutive patients referred for a psychiatric evaluation in an urban ED, symptom severity was quantified using the Brief Psychiatric Rating Scale (BPRS). The BPRS scores of all non-suicidal patients were subjected to receiver operator characteristic (ROC) curve analysis to determine the sensitivity, specificity, and optimal cutoff score of the BPRS in predicting admission for non-suicidal patients. RESULTS: A BPRS cutoff score of 39 had a sensitivity of 85.71 percent and a specificity of 86.11 percent. The area under the ROC curve was .8671 (Somer's D = .7342) and the standard error of the curve was .1124. The cutoff score of 39 correctly identified six of seven non suicidal patients who were hospitalized. CONCLUSIONS: It is anticipated that use of the BPRS in the ED will be further refined when a larger patient sample is studied. Potentially, a subset of BPRS items could be identified which would be more sensitive in predicting admission than the full BPRS and increase the overall efficiency of administering the BPRS in the ED. PMID- 9724892 TI - Laparoscopic and psychologic evaluation of women with chronic pelvic pain. AB - OBJECTIVE: Pelvic pain can account for up to 40 percent of laparoscopies performed by gynecologists. This report compares the psychological profiles and efficacy of laparoscopic surgery at long-term follow-up in a series of laparoscopy-positive and laparoscopy-negative patients with chronic pelvic pain. METHOD: A retrospective chart review was performed on patients diagnosed with chronic pelvic pain combined with postoperative written questionnaires and self rating scales. These questionnaires were used to assess long-term post laparoscopy follow-up of the physical and psychological status of women with positive findings at laparoscopy compared to those women with negative findings. RESULTS: There were no statistically significant demographic differences between respondents and nonrespondents. In the respondents, no statistically significant differences were noted even with long-term follow-up when comparing responses of the laparoscopy-positive and laparoscopy-negative groups on the above questionnaires. CONCLUSION: Though reporting modest improvement in pelvic pain since laparoscopy, both groups reported a high incidence of anxiety, depression, physical worries, and marital/sexual problems. PMID- 9724893 TI - Risperidone: treatment response in adult and geriatric patients. AB - OBJECTIVE: To compare the efficacy and side effects of risperidone in younger adult and geriatric patients. METHODS: Open retrospective study of 102 consecutive intakes, prescribed risperidone, by a mental health team. All patients were non-hospitalized community residents. Prior to initiation of risperidone, and at termination of study period, Clinical Global Impression (CGI) scores were used to track progress. Variables monitored were: concurrent use of other antipsychotics, compliance, side effects, and maintenance dosage. RESULTS: The most common DSM-IV diagnoses were schizophrenia in the younger adult group and late onset delusional disorders in the geriatric group. Compliance was good for both groups. The geriatric group demonstrated a greater treatment response which was reached at a significantly lower dosage. There was no statistically significant difference in the occurrence of side effects. Examination of response by diagnostic category indicated that geriatric patients with late onset delusional disorder showed the best response while adults with either schizophrenia or affective syndromes also showed positive response. CONCLUSIONS: Risperidone, at lower than recommended doses, shows promise in the treatment of late onset delusional disorders and behavior syndrome of dementia. The side effect profile was benign, as was suggested by experience in treating schizophrenia. Scientifically more rigorous prospective studies for the indications and efficacy of risperidone in late onset psychotic disorders and psychoses and behavior syndromes associated with dementing illness are overdue. PMID- 9724894 TI - Polymer surface chemistry and bone cell migration. AB - Implant devices for orthopaedic applications may be improved if the surface of the biomaterial provides for osteointegration. To understand the effect of hydrophilicity on colonisation by human bone derived (HBD) cells, we compared untreated polystyrene (PS) and a sulfuric acid-treated PS surface for mechanisms of cell migration. The chemical composition of the acid-treated PS surface was analysed by monochromatic X-ray photoelectron spectroscopy and found to contain various oxidatively produced groups and a minor amount of sulfonate groups. It was found that migration of HBD cells on both PS and acid-treated PS surface was dependent on the presence of vitronectin (Vn) and was higher on the hydrophilic acid-treated surface. Minimal migration of HBD cells occurred on either surface in the absence of Vn, even when fibronectin was present in the culture medium. Using radiolabelled protein, it was shown that Vn adsorption onto the acid treated surface was two to three fold greater than that on the hydrophobic PS. When HBD cells were seeded onto a patterned surface in a medium containing Vn, the cells preferentially colonised the hydrophilic region and few, if any, cells traversed the haptotactic boundary from the hydrophilic to the hydrophobic side. Thus the enhanced HBD cell migration seen on the acid-treated PS compared with the untreated PS surface and the haptotactic boundary phenomenon, relate to Vn adsorption. PMID- 9724895 TI - Platelet adhesion on the gradient surfaces grafted with phospholipid polymer. AB - We have synthesized omega-methacryloyloxyalkyl phosphorylcholine (MAPC) polymers as new blood-compatible materials, with attention to the surface structure of the biomembrane and investigated their blood compatibility. The blood compatibility observed on the MAPC polymers is due to their strong affinity to phospholipids. When the blood comes in contact with the MAPC polymer, phospholipids in the plasma preferentially adsorb on the surface, compared with the plasma proteins or cells. The adsorbed phospholipids construct a biomembrane-like structure on the MAPC polymer surface. The MAPC polymers then have an excellent blood compatibility. In this study, we prepared a gradient poly(MAPC)-grafted polyethylene (PE) surface using a corona discharge treatment method to clarify the effect of the chemical structure of the MAPC unit on the blood compatibility of the MAPC polymers. The surface composition of MAPC and the hydrophilicity on the poly(MAPC)-grafted PE surface were determined by X-ray photoelectron spectroscopic (XPS) analysis and contact angle measurement with water, respectively. The phosphorus/carbon (P/C) ratio determined by the XPS analysis increased, but the water contact angle decreased with increasing corona irradiation energy. These results indicated that the surface density of the MAPC unit was increased. More than 2.5 cm from the starting point of the corona irradiation, the P/C ratio and water contact angle of the surface achieved a constant level. Thus, the surface was completely covered with the grafted poly(MAPC) chain. The effect of the methylene chain length of the MAPC unit on surface properties was also observed. The phospholipid polar group of the MAPC unit was effectively exposed on the surface as the chain length became longer. Moreover, the hydrophobicity of the surface was increased with the increase in the methylene chain length of the MAPC unit. The number of platelets adhering to the poly(MAPC)-grafted PE surface was reduced from the same point where the P/C ratio became constant. PMID- 9724896 TI - Specific activation of platelets by surface-adsorbed plasma proteins. AB - Platelets were isolated from human blood by Percoll density gradient centrifugation in a low Ca2+/high Mg2+ buffer. The buffer reversibly inactivates the cells during separation. The purity of the isolated cells (> 99%) was determined by flow cytometry, their viability was confirmed by fluorescein diacetate hydrolysis, and their morphology was studied with TEM. Plasma proteins were adsorbed onto hydrophobic glass surface, and pure platelets were added and incubated for up to 30 min at 37 degree C. Platelet activation was determined by cell spreading, formation of microparticles and surface exposure of CD62P indicating the release of alpha-granules. Surface-immobilized IgG was shown to cause the release of microparticles and cell lysis, in accordance with data published by others. Surface-immobilized vWF was shown to induce CD62P exposure on the platelet cell surface. The specificity of this response was demonstrated by adsorbing plasma proteins from normal and factor VIII-deficient plasma. PMID- 9724897 TI - Macrophage--polymer interactions. AB - One of the most notable and initial indications of the body's response to an implanted material is the inflammatory response, a process which is known to be largely influenced by the activation of macrophages and other cell types. Thus, the magnitude of the inflammatory response can be related to the level of activation of macrophages. As certain well defined morphological changes are known to accompany cell activation, we quantitatively evaluated the cell- substrate interactions, between a range of polymeric materials and isolated macrophages in vitro, using microscopy and image analysis. This enabled us to assess the morphology of the cells, and how the morphology of the macrophages was influenced by changes in time and substrate. This method provided detailed images which were used to evaluate cell-substrate interactions. It was found that, polyurethane (PU) and poly(dimethylsiloxane) (Si) samples displayed a similar pattern in cell behaviour, whilst macrophages placed in contact with polyvinylchloride (PVC) samples continually displayed a more activated morphology with increasing time throughout the test period. As well as providing a detailed analysis of cell-substrate interactions this study also highlighted the advantages of employing an image analysis program for a more comprehensive and reproducible form of assessment of cellular morphology. PMID- 9724898 TI - The modulation of cellular responses to poly(2-hydroxyethyl methacrylate) hydrogel surfaces: phosphorylation decreases macrophage collagenase production in vitro. AB - We examined the regulation of collagenase production by the monocyte/macrophage THP-1 cell line when these cells were exposed to poly(2-hydroxyethyl methacrylate) (PHEMA) hydrogel surfaces with different chemistries and morphologies. Tissue culture modified polystyrene (TCP), used as a control surface, induced the maximum collagenase response. Copolymer hydrogels containing 2-ethoxyethyl methacrylate (EMA) or methyl methacrylate (MMA) also induced a high response, while PHEMA hydrogels induced a low level response and the phosphorylated hydrogel induced no response. This pattern was altered when the morphology of the hydrogels was changed to that of a sponge. The overall enzyme response to the sponge hydrogels was lower than that to the homogeneous hydrogels. Sponges containing EMA and MMA produced low level response relative to the TCP control. PHEMA and phosphorylated sponges produced little and no response respectively. The dramatically reduced enzyme response to phosphorylated surfaces was not a consequence of cell death, and may be a phenomenon related to changes in cell surface charge. PMID- 9724899 TI - Small intestinal submucosa: a substrate for in vitro cell growth. AB - The extracellular matrix (ECM) of the small intestinal submucosa (SIS) was harvested by removing the superficial layers of the mucosa and the external muscular layers. The remaining 80 microns thick sheet was disinfected and sterilized by methods which removed all cellular components. The SIS-ECM, retaining its native 3-dimensional microarchitecture and composition, was evaluated for its ability to support in vitro cell growth. Six separate cell types were seeded either alone or in coculture with other cells upon this matrix, grown in selected media, a examined daily for time periods ranging from 48 h to 2 weeks. The six cell types tested were NIH Swiss mouse 3T3 fibroblast, NIH 3T3/j2 fibroblasts, primary human fibroblasts, primary human keratinocytes, human microvascular endothelial cells (HMECs), and an established rat osteosarcoma (ROS) cell line. All cell types showed the ability to attach a proliferate. All fibroblast cell line and the keratinocytes proliferated and/or migrated into the 3-dimensional scaffold of the SIS matrix. The ROS cells and the HMECs were confined in their growth pattern to the surface of the matrix. Coculturing of NIH 3T3/J2 fibroblasts and primary human keratinocytes resulted in a distinctive spatial orientation of the two cell types. The fibroblast populated the mid substance of the 3-dimensional matrix and the keratinocytes formed an epidermal structure with rete ridge-like formation and stratification when the composite was lifted to an air liquid interface in culture. In summary, SIS provides a substratum with a 3-dimensional scaffold that allows for cell migration and spatial organization. The substratum is suitable for in vitro studies of the interaction between epithelial or mesenchymal cells and a naturally occurring extracellular matrix. PMID- 9724900 TI - Artificial juxtacrine stimulation for tissue engineering. AB - To endow biomaterials with the ability to regulate cell functions such as proliferation, differentiation, and apoptosis, growth factor proteins were covalently immobilized. The proteins were immobilized on various matrices using different chemical methods. It was shown that insulin and epidermal growth factor stimulated cellular functions even after immobilization. Pattern-immobilization of growth factor proteins clearly demonstrated the stimulation by immobilized proteins. In other words, this type of stimulation by non-diffusional growth factors enabled us to regulate tissue formation with artificial biomaterials. The stimulation was enhanced by coimmobilization with adhesion factors. These stimulations due to the immobilized growth factors may mimic juxtacrine stimulation of membrane-anchored growth factors such as heparin-binding epidermal growth factor, transforming growth factor-alpha, and tumor necrosis factor-alpha. PMID- 9724901 TI - Fertility regulation in cattle. AB - This paper reviews the physiological, endocrinological and pharmaceutical literature pertaining to the design, development and optimisation of subcutaneous and intravaginal progestogen-containing drug delivery systems used in the control of synchrony and ovulation in cattle. PMID- 9724902 TI - Proniosome based transdermal delivery of levonorgestrel for effective contraception. AB - A proniosome based transdermal drug delivery system of levonorgestrel (LN) was developed and extensively characterized both in vitro and in vivo. The proniosomal structure was liquid crystalline-compact niosomes hybrid which could be converted into niosomes upon hydration. The system was evaluated in vitro for drug loading, rate of hydration (spontaneity), vesicle size, polydispersity, entrapment efficiency and drug diffusion across rat skin. The effect of composition of formulation, amount of drug, type of Spans, alcohols and sonication time on transdermal permeation profile was observed. The stability studies were performed at 4 degrees C and at room temperature. The biological assay for progestational activity included endometrial assay and inhibition with the formation of corpora lutea. The study demonstrated the utility of proniosomal transdermal patch bearing levonorgestrel for effective contraception. PMID- 9724903 TI - In vitro and in vivo release of ciprofloxacin from PLGA 50:50 implants. AB - Poly(lactides-co-glycolides) [PLGA] are widely investigated biodegradable polymers and are extensively used in several biomaterials applications as well as drug delivery systems. These polymers degrade by bulk hydrolysis of ester bonds and break down into their constituent monomers, lactic and glycolic acids which are excreted from the body. The purpose of this investigation was to develop and characterize a biodegradable, implantable delivery system containing ciprofloxacin hydrochloride (HCl) for the localized treatment of osteomyelitis and to study the extent of drug penetration from the site of implantation into the bone. Osteomyelitis is an inflammatory bone disease caused by pyogenic bacteria and involves the medullary cavity, cortex and periosteum. The advantages of localized biodegradable therapy include high, local antibiotic concentration at the site of infection, as well as, obviation of the need for removal of the implant after treatment. PLGA 50:50 implants were compressed from microcapsules prepared by nonsolvent-induced phase-separation using two solvent-nonsolvent systems, viz., methylene chloride-hexane (non-polar) and acetone-phosphate buffer (polar). In vitro dissolution studies were performed to study the effect of manufacturing procedure, drug loading and pH on the release of ciprofloxacin HCl. The extent of penetration of the drug from the site of implantation was studied using a rabbit model. The results of in vitro studies illustrated that drug release from implants made by the nonpolar method was more rapid as compared to implants made by the polar method. The release of ciprofloxacin HCl. The extent of the penetration of the drug from the site of implantation was studied using a rabbit model. The results of in vitro studies illustrated that drug release from implants made by the nonpolar method was more rapid as compared to implants made by the polar method. The release of ciprofloxacin HCl from the implants was biphasic at < or = 20% w/w drug loading, and monophasic at drug loading levels > or = 35% w/w. In vivo studies indicated that PLGA 50:50 implants were almost completely resorbed within five to six weeks. Sustained drug levels, greater than the minimum inhibitory concentration (MIC) of ciprofloxacin, up to 70 mm from the site of implantation, were detected for a period of six weeks. PMID- 9724904 TI - Equilibrium swelling of poly(methacrylic acid-g-ethylene glycol) hydrogels. Effect of swelling medium and synthesis conditions. AB - Responsive hydrogel networks comprising of poly(methacrylic acid) (PMAA) backbone and oligomeric ethylene glycol (PEG) grafts were synthesized by free-radical solution polymerization and their equilibrium swelling properties were characterized in aqueous solutions of a homologous series of alcohols. These hydrogels are known to exhibit swelling transitions in response to external stimuli which lead to formation or disruption of hydrogen-bonded complexes between the backbone and the grafts. Swelling studies performed in aqueous mixtures of methanol, ethanol and propanol revealed that the effectiveness of an alcohol in breaking the PMAA/PEG complexes increased as the aliphatic segment length of the alcohol was increased. These results confirm the importance of hydrophobic interactions for stabilizing the complexes. Studies performed to determine the effect of the synthesis conditions on the equilibrium swelling properties revealed that the equilibrium degree of swelling increased as the solvent fraction during synthesis was increased. Finally, molecular stimulations revealed that it is sterically possible to form complexes with a 1:1 stoichiometry between chains of poly(methacrylic acid) and poly(ethylene glycol) with essentially no additional bond strain. PMID- 9724905 TI - Formulation and in vitro evaluation of ibuprofen-Carbopol 974P-NF controlled release matrix tablets. III: Influence of co-excipients on release rate of the drug. AB - In order to assess the potential of carbopol 974P-NF as matrix material in hydrophilic matrix tablets containing a slightly water-soluble drug, ibuprofen (IBF), controlled release matrix tablets of ibuprofen and carbopol 974P-NF, at different drug to polymer ratios, were prepared by the direct compression method. The influence of the concentration of the matrix material (carbopol 974P) and several co-excipients (lactose, microcrystalline cellulose, and starch) on the release rate of the drug was investigated. An in vitro dissolution test in pH 7.2 phosphate buffer solution showed that drug release from all the formulations containing carbopol 974P was considerably prolonged in concentration-dependent manners. Increasing the amount of carbopol 974P in tablets resulted in a reduction in the drug release rate and a linearization of the drug release curve. When the influence of the co-excipients on the release of the drug was examined, all of the co-excipients used in this study enhanced the release rate of IBF. However, lactose demonstrated slower and more linear release behavior as compared in microcrystalline cellulose or starch. The dissolution T50 and T90 values for the three co-excipients were in the order of lactose > microcrystalline cellulose > starch. PMID- 9724906 TI - Dual-stimuli-responsive drug release from interpenetrating polymer network structured hydrogels of gelatin and dextran. AB - Interpenetrating polymer network (IPN)-structured hydrogels of gelatin (Gtn) and dextran (Dex) were prepared with lipid microspheres (LMs) as a drug microreservoir, and LM release from these hydrogels was examined in relation to their dual-stimuli-responsive degradation. A phase morphology in the IPN structured hydrogels was varied with the preparation temperature, i.e. above or below the sol-gel transition temperature (Ttrans) of Gtn. The IPN-structured hydrogel prepared below Ttrans exhibited a specific degradation-controlled LM release behavior: LM release from the hydrogel in the presence of either alpha chymotrypsin or dextranase alone was completely hindered, whereas LM release was observed in the presence of both enzymes. It is concluded that dual-stimuli responsive drug release can be achieved by specific degradation of a particular IPN-structured hydrogel. PMID- 9724907 TI - Arterial uptake of biodegradable nanoparticles for intravascular local drug delivery: results with an acute dog model. AB - Biodegradable nanoparticles (NP) with a spherical diameter ranging from 70 to 160 nm were investigated for potential usefulness for the local intraluminal therapy of restenosis, the disease process responsible for arterial reobstruction following angioplasty. NPs containing a water-insoluble anti-proliferative agent U-86983 (U-86, Pharmacia and Upjohn, Kalamazoo, MI) were formulated from oil water emulsions using biodegradable polymers such as poly(lactic acid-co-glycolic acid) (PLGA), and specific additives after particle formation, to enhance arterial retention using either heparin, didodecylmethylammonium bromide (DMAB), or fibrinogen, or combinations. Femoral and carotid arteries of male mongrel dogs were isolated in situ, and were then subjected to a balloon angioplasty. A NP suspension of a predetermined concentration was then infused into the artery for various durations. This was followed by a 30 min restoration of blood flow through the vessel. The arterial segments were excised and analyzed for drug levels. From the drug loading the NP and the drug levels in the artery, the quantity of nanoparticles retained was calculated and expressed as microgram per 10 mg dry arteries. In general, repeated short infusions of nanoparticle suspension (15 s x 4) were two-fold more effective in terms of higher arterial U 86 levels than a single prolonged infusion (60 s). A single 15 s infusion was not significantly different than a 60 s compared to non-modified NPs (39.2 +/- 2.5 and 49.1 +/- 2.4 vs. 21.5 +/- 0.6 micrograms/10 mg mean +/- s.e., respectively). A comparably enhanced NP uptake was noted with a combined heparin/DMAB modification. Increasing the concentration of NP in infusate from 5 to 30 mg ml-1 significantly increased arterial NP uptake level (from 22.5 +/- 3.5 to 83.7 +/- 1.4 micrograms/10 mg). Thus, the results support the view that modified nanoparticles along with optimized infusion conditions could enhance arterial wall drug concentrations of agents to treat restenosis. PMID- 9724908 TI - Release of enzymes from liposomes during cheese ripening. AB - Changes in proteolysis and in residual enzymatic activity as a function of time were compared in model cheeses, made with either free enzymes or liposomes containing enzymes and in control model cheeses. Cheeses were ripened under different conditions of pH, fat content and temperature. The release of enzymes from liposomes was significantly stimulated by increasing the fat content from 0 to 20% and the pH from 4.9 to 5.5. Ripening temperature (6 degrees C or 13 degrees C) did not affect 2 months of ripening, proteolysis was 30% lower in liposome-than in free enzyme-treated cheeses, indicating a possible inhibition of released enzymes. PMID- 9724910 TI - Recent advances in evolutionary psychology and psychiatry. PMID- 9724909 TI - HPMA copolymer bound adriamycin overcomes MDR1 gene encoded resistance in a human ovarian carcinoma cell line. AB - N-(2-Hydroxypropyl)methacrylamide (HPMA) copolymer-adriamycin (ADR) conjugate containing lysosomally degradable oligopeptide (GFLG) side chains terminated in ADR was synthesized. The effect of free and HPMA copolymer-bound ADR on the viability of A2780 sensitive and A2780/AD multidrug resistant human ovarian carcinoma cells was studied in vitro. As expected, the IC50 dose for the HPMA copolymer-ADR conjugate was higher than for free ADR reflecting the difference in the mechanism of cell entry. The resistant A2780/AD cells demonstrated about 40 times higher resistance to free ADR than the sensitive A2780 cells. On the contrary, there was only a small difference in cytotoxicity of the HPMA copolymer ADR conjugate toward sensitive A2780 or MDR resistant A2780/AD cells. The IC50 value for A2780/AD was only about 20% higher than the value for sensitive A2780 cells. These data seem to indicate that the HPMA copolymer-ADR conjugate may, at least partially, avoid the ATP driven P-glycoprotein (Pgp) efflux pump. The analysis of the expression of the MDR1 gene which encodes the Pgp, has shown that free ADR in high doses stimulated MDR1 gene expression in sensitive A2780 cells. At the same time both free and HPMA copolymer-ADR conjugate partially inhibited the expression of the MDR1 and beta 2 m genes in multidrug resistant A2780/AD cells. PMID- 9724911 TI - The "ideologic" view of transference. PMID- 9724912 TI - Freud and castration: a new look into the origins of psychoanalysis. PMID- 9724913 TI - The metaphor of time in psychoanalytic technique. PMID- 9724914 TI - Some thoughts concerning borderline pathology and fear of aloneness. PMID- 9724915 TI - Enactments that don't appear as enactments. PMID- 9724916 TI - The unexplored complementarity of short-term and long-term analytic approaches. PMID- 9724917 TI - The integration of sensorial channels through progressive mirror drawing in the psychotherapy of schizophrenic patients with disturbances in verbal language. PMID- 9724918 TI - "Like those crystal balls of yore..." fragility within the analytic relation: real or fancied? PMID- 9724920 TI - All about Barbie: distortions of a transitional object. PMID- 9724919 TI - Kleinian guilt, determinism, and free will: implications for clinical theory and treatment. AB - The purpose of this discussion of teleological and deterministic concepts in clinical theory and treatment was not to offer definitive answers to a timeless philosophical problem. Indeed, a solution to the problem of free will in a deterministic science eludes even the greatest thinkers (cf. Howard and Conway, 1987). Following Feyerabend (1984), the purpose was instead to question one of the "general and lasting boundary conditions" (p.3)-causal explanations introduced by Enlightenment-era philosophers of science to "protect" (p.3) science. I have attempted to demonstrate the logical inconsistencies of Klein's theory of etiology of infantile guild using both a teleological perspective that questions the assumption of causal explanations imposed on a scientific inquiry of human experience, and a deterministic perspective that affirms this assumption. When taken to their logical conclusions, both perspectives reduce to mutually irreconcilable theoretical and clinical absurdities. As as clinician I myself have chosen to work within a deterministic framework. I have found that perceiving the patient deterministically allows me to empathize with even the most unspeakable intrapsychic experiences. I share with my patient the antecedent conditions that efficiently cause our common experiences. It is this empathy for our share human condition-in al its transcendence, frailty, and profanity communicated through interpretation that seems to give birth to the healing process (Goodman, 1991). This discussion doubtless also contains inconsistencies. Nevertheless, it contributes to the ongoing discourse of clinical theory and treatment because it draws attention to the enormous, yet still largely implicit, debt that clinical theory and treatment owe to philosophy. It is a great irony that the discipline that has historically put scientific inquiry in shackles also has the power to set it free. Whether teleology is the philosophical perspective to set clinical theory and treatment free, however, seems unlikely. In choosing to discuss Klein's theory of infantile guilt, I have taken seriously R.J. Rychlak and Rychlak's (1990) implicit support (cf. p.48) for a teleology that functions throughout the life span, et find it difficult to imagine both the unconscious and conscious minds with freely premising capacities-as if human being were actually two persons in one. One the other hand, reducing human beings to the status of Hobbesian machines-as the deterministic perspective seems to imply presents a similarly unattractive alternative. To understand the nature of human suffering and the process of healing it, however, one thing is certain: A philosophical discourse of clinical theory and treatment must continue. PMID- 9724921 TI - International Congress of Gynecologic Endoscopy, 27th annual meeting of the American Association of Gynecologic Laparoscopists. Atlanta, Georgia, USA. November 10-15, 1998. Abstracts. PMID- 9724922 TI - The intracellular potassium and chloride channels: properties, pharmacology and function (review). AB - Channels selective for potassium or chloride ions are present in membranes of intracellular organelles such as sarcoplasmic (endoplasmic) reticulum, mitochondria, nucleus, synaptic vesicles, and chromaffin, and zymogen granules. They probably play an important role in cellular events such as compensation of electrical charges during transport of Ca2+, delta pH formation in mitochondria or V-ATPase containing membrane granules, and regulation of volume changes, due to potassium and chloride transport into intracellular organelles. Intracellular potassium and chloride channels could also be the target for pharmacologically active compounds. This mini-review describes the basic properties, pharmacology, and current hypotheses concerning the functional role of intracellular potassium and chloride channels. PMID- 9724923 TI - The modulation of membrane structure and stability by dimethyl sulphoxide (review). AB - Dimethyl sulphoxide is a widely used agent in cell biology. It is well known as a cryoprotectant, cell fusogen and a permeability enhancing agent. These applications depend, to a greater or lesser extent, on the effect of dimethyl sulphoxide on the stability and dynamics of biomembranes. The aim of this review is to examine progress of the research which has been directed towards studies of the interactions between dimethyl sulphoxide and membranes, particularly that with the lipid components of cell membranes, as seen in its effects on model membrane systems. Models are proposed to explain the mechanism whereby dimethyl sulphoxide may mediate its effects on biological functions by its effects on the stability and properties of the membrane lipid matrix. PMID- 9724924 TI - Microcalorimetric study on the phase behaviour of S-layer coated liposomes. AB - Isolated S-layer subunits from Bacillus coagulans E38-66/v1 were recrystallized on positively charged, unilamellar liposomes composed of dipalmitoylphosphatidylcholine, cholesterol and hexadecylamine. The thermotropic phase behaviour of S-layer coated and uncoated liposomes was characterized by differential scanning microcalorimetry indicating for both preparations a broad transition around 50 degrees C due to the chain-melting from a liquid-ordered gel like to a liquid-ordered fluid phase as described for phosphatidylcholine/cholesterol mixtures. The slightly higher phase transition temperature for the S-layer coated liposomes were explained by increased intermolecular order. Cross-linking the S-layer subunits covalently to hexadecylamine with glutaraldehyde induced phase separation within the liposomes. Based on deconvolution of the normalized excess heat capacity functions it was proposed that the different lipid domains arise from phospholipids representing different degrees of mobility. PMID- 9724925 TI - Assignment of the human serotonin 4 receptor gene (HTR4) to the long arm of chromosome 5 (5q31-q33). AB - In the present study, we report the chromosomal localization of the human 5-HT4 receptor gene (HTR4) by the analysis of somatic cell hybrids. Based upon genomic sequences of the HTR4 gene, a primer set was selected that reacted with human genomic DNA but not mouse or hamster genomic DNA. Using monochromosomal hybrid cell lines of the NIGMS Mapping Panel 2 we localized the HTR4 gene to human chromosome 5. To confirm the localization on chromosome 5 and to further sublocalize the HTR4 gene, the radiation hybrid panel RH3 was similarly analysed. Significant linkage was obtained to genetic marker D5S2654. This localizes the HTR4 gene to human chromosome 5q31-q33. PMID- 9724926 TI - Expression of the mammalian renal peptide transporter PEPT2 in the yeast Pichia pastoris and applications of the yeast system for functional analysis. AB - It has recently been identified the PEPT2 cDNA encodes the high affinity proton coupled peptide transporter in rabbit kidney cortex. PEPT2 represents a 729 amino acid protein with 12 putative transmembrane domains that mediates H+/H3O+ dependent electrogenic transmembrane transport of di- and tripeptides and of selected peptidomimetics. Here the functional expression of PEPT2 in the methylotropic yeast Pichia pastoris is described under the control of a methanol inducible promoter. Western blot analysis of Pichia cell membranes prepared from a recombinant clone identified a protein with an apparent molecular mass of about 85-87 kDa. Peptide uptake into cells expressing PEPT2 was up to 80 times higher than in control cells. Cells of recombinant clones showed a saturable peptide transport activity for the hydrolysis resistant dipeptide 3H-D-Phe-Ala with an app. K0.5 of 0.143 +/- 0.016 mM. Inhibition of 3H-D-Phe-Ala uptake by selected di and tripeptides and beta-lactam antibiotics revealed the same substrate specificity as obtained in renal membrane vesicles or for PEPT2 when expressed in Xenopus laevis oocytes. A novel fluorescence based assay for assessing transport function based on a coumarin-labeled fluorescent peptide analogue has also been developed. Moreover, using a histidyl auxotrophe strain a PEPT2 expressing cell clone in which transport function can be monitored by a simple yeast growth test was established. In conclusion, this is one of only a few reports on successful functional expression of mammalian membrane transport proteins in yeast. The high expression level will provide a simple means for future studies either on the structure-affinity relationship for substrate interaction with PEPT2 or for selection of mutants generated by random mutagenesis. PMID- 9724927 TI - Amphiphile-induced phosphatidylserine exposure in human erythrocytes. AB - Nonionic and anionic water-soluble amphiphiles were shown to increase strongly the binding of fluorescein isothiocyanate-conjugated annexin V (FITC-annexin V) in human erythrocytes pretreated with the aminophospholipid translocase (APLT) inhibitor n-ethylmaleimide (NEM). At high sublytic amphiphile-concentrations the binding of FITC-annexin V, monitored in a flow cytometer, was time- and temperature-dependent and occurred heterogeneously in the cell population, with 43-81% of cells being stained above background following incubation for 60 minutes at 37 degrees C. The increased FITC-annexin V binding apparently indicates an increased flop rate of phosphatidylserine (PS) to the outer membrane leaflet. When the NEM-pretreatment was omitted, the FITC-annexin V binding was markedly, but not completely, reduced. In erythrocytes incubated with a zwitter ionic amphiphile, a small increase in FITC-annexin V binding was detected, while cationic amphiphiles did not induce an increased FITC-annexin V binding. The potency of amphiphiles to induce PS exposure was not related to the type of shape alteration or vesiculation induced. Our results indicate a significant role of the charge status of a membrane intercalated amphiphile for its capability to induce PS exposure. PMID- 9724928 TI - Heterosexually acquired HIV infection in Doncaster. PMID- 9724929 TI - Salmonella food poisoning linked to north London bar mitzvah. PMID- 9724931 TI - Drugs and the elderly. PMID- 9724930 TI - The latest on irritable bowel syndrome. PMID- 9724932 TI - Taking the threat out of threatening questions. AB - A major threat to the validity and reliability of health and social surveys is that questions asked are regarded as threatening. This leads to the unpredictability of social desirability bias, which can skew responses in a variety of directions. The types of situations in which this may occur are outlined, together with a ten point approach that will assist in overcoming the problems. PMID- 9724933 TI - Secondary prevention of stroke--an update. AB - Stroke disease remains a national priority due to the high morbidity and mortality. Good clinical practice dictates that risk factors are identified and corrected. Various therapeutic regimens have been tested in trials of sufficient strength to give answers. In the second European Stroke Prevention Study (ESPSZ), low dose aspirin (50 mg/d), sustained release dipyridamole (200 mg/d) were shown to reduce recurrence of stroke and TIA. The combination was twice as effective. Also in the Clopidogrel/aspirin Prevention of Recurrence of Ischaemic Events (CAPRIE) trial clopidogrel was shown to be superior to aspirin. In clinical practice patients with stroke or TIA should be investigated rapidly, their risk factors corrected and appropriate drug regimens implemented. PMID- 9724934 TI - Antiseptics: a forgotten weapon in the control of antibiotic resistant bacteria in hospital and community settings? AB - The aim of this study was to ascertain the activity of a selection of widely-used antiseptic/disinfectant agents against antibiotic resistant bacteria and strains isolated from patients infected with clinically significant species. Four antiseptic agents (Dettol, Dettol Hospital Concentrate, Savlon and Betadine) were tested against Staphylococcus aureus, Methicillin Resistant S. aureus (MRSA), Enterococcus hirae, Vancomicin Resistant Enterococcus sp (VRE), Escherichia coli and E. coli 0157. The antiseptics were applied at recommended use dilutions and at a half and a quarter of those concentrations in a standard suspension test (EST). Organic material was added to mimic the presence of blood, protein and other such contaminants to be found in the clinical situation. All antiseptics tested were effective against both the antibiotic sensitive and antibiotic resistant strains of S. aureus and E. hirae as well as normal and clinical strains of E. coli at recommended concentrations. All but Betadine were also effective against the antibiotic resistant bacteria at a half and a quarter of normal concentration. The iodine containing antiseptic, however, failed the test against MRSA at a half normal concentration and showed virtually no activity against MRSA at a quarter normal concentration. PMID- 9724935 TI - The Food Standards Agency: a force for change. PMID- 9724936 TI - Pharmacy involvement in emergency preparedness/response. PMID- 9724937 TI - Decreasing requirement for lithium carbonate therapy in bipolar affective disorders (hypomanic type) following the onset of chronic renal insufficiency. AB - The importance of regular monitoring of both serum lithium and creatinine levels, together with thyroid function assessment, in a patient taking lithium carbonate therapy for bipolar affective disorder (hypomanic type) is emphasised. In this case it was the gradual rise of serum creatinine that alerted the physician to the onset of insidiously progressive renal impairment. In the absence of any evidence for another aetiology, it was concluded that a possible cause for the renal problem was the lithium therapy itself. By reducing the dosage it was found that serum lithium levels were maintained within the reference range, thus avoiding the potential psychiatric consequences of high concentrations--which could well have occurred had the former dosage been continued during the period of deteriorating renal function. The situation is currently being carefully monitored in case another cause for renal disease, other than that of a side effect of therapy, emerges at a later date. PMID- 9724938 TI - Changes in the numbers of dentists and dental caries levels in 12-year-olds in the countries of the European Union and economic area. AB - This study compares numbers of dentists in the countries of the European Union (EU) from 1970 to 1994 with dentist to population ratios and dental caries levels in 12-year-olds, sets the changes which have emerged against other changing dental disease patterns (including those unconnected with caries levels) and goes on to determine the significance of the findings to the problems of dental workforce planning. Data for the numbers of dentists and dentist to population ratios were obtained from published tables. Data for past caries levels in 12 years-olds were obtained from the WHO Global Data Bank. All other data were obtained from Chief Dental Officers in all countries of the EU. In 1994 there were 222,090 practising dentists in the EU and 12,853 other clinical workers of whom 11,493 were dental hygienists. Since 1970, the dentist:population ratios for Spain and Portugal have improved markedly, the improvement for other countries has been less marked and in Austria a small reduction has occurred. Six countries show a considerable reduction in caries levels for 12-year-olds between 1970 and the 1990s, five show smaller reductions and three show a deterioration. As a consequence of the general improvement in caries levels in most of these countries it is probable that workloads in relation to the treatment of caries are falling, particularly for younger age groups. However, because of the overall ageing of populations in many industrialised countries the workload for older age groups is increasing, as older adults increase in number, a greater proportion retain their teeth and are afflicted by a range of problems, which include, but are not exclusive to, dental caries. There are few data for these older age groups than for caries levels in 12-year-olds. It was concluded that those planning the EU dental workforce of the future should take account of reliable epidemiological data for all groups of the population and, as these are not currently available, that suitable periodic oral health surveys covering all population age groups should be carried out regularly in all EU member states. PMID- 9724939 TI - Community care: fracturing managerial frames. AB - Managers come to their task with their frames of reference--certain assumptions- a mind set. They wish to be seen as rational and consistent; to live rent-free in their comfort zones. There is the lure of 'Social Proof'; to act like others placed in a similar situation. Deviance is seen as counter-cultural. This approach is no longer tenable; the antiquity of an error is no reason for its continuance. The entrepreneurial manager must move from the concept of 'fit'- resources to match objectives--to one of 'stretch', which argues for a misfit between strategy and resources; between funding and aspirations. One must do more with less. PMID- 9724940 TI - Bile acids, fibre and colon cancer: the story unfolds. AB - Are the changes in faecal bile acid concentrations the cause of colorectal cancer or one of its effects? This is an area of controversy mainly due to the lack of a clear explanation as to how the bile acid concentrations are controlled under different circumstances. This review presents an outline of the evidence that bile acids are both a causal factor in colorectal cancer and that their concentrations are affected by it in a synergistic manner. It also offers an explanation of how some dietary fibre protects against colorectal cancer. PMID- 9724941 TI - NHS reorganisations. PMID- 9724942 TI - 'Not too much, not too little, but just enough?': observations on continuing professional development in Public Health in the north of England. AB - Three inquiries about Public Health continuing professional development were undertaken in the Northern and Yorkshire Region of the National Health Service during 1995-96. Public Health Physicians were asked about their experience of continuing education and for their views on a regional policy for continuing professional development. Health Authority Chief Executives were asked about their reactions to Public Health Physicians continuing educational needs. The overall response rates for the Public Health Physicians were very disappointing. Most of the Chief Executives (a much smaller group) responded to the inquiry. A large minority of Public Health Physicians believed that their continuing education in the preceding two years had been adequate. Most wished their future continuing education activities to be multi-disciplinary. One finding with considerable significance for those managing Public Health education, both specialist and continuing, was that many of those with teaching responsibilities had not been trained to teach. Despite apparent concordance between the views of Chief Executives and those of Public Health Physicians; on some important points there were inconsistencies in the comments of Chief Executives, which suggested lack of understanding of both the roles of their professional colleagues and the need for their continuing education. The inquiries gave rise to a sense of apathy and under-confidence, manifested in some reluctance to accept policing of continuing education. There is a need to experiment with learning and teaching approaches in order to progress from the current traditional educational methods. PMID- 9724943 TI - Research, ethics and privacy: the limits of knowledge. AB - This paper considers the need for personal privacy within the context of epidemiological research. It concludes that privacy can be protected by early anonymization and aggregation of personal health data without prejudicing the viability of a research project. During the period before anonymization, however, a secure legal framework is necessary to prevent unauthorized access to potentially sensitive information. Within such a framework ethical codes need to be identified and monitored by an appropriate Local Research Ethics Committee (or a Multicentre Ethics Committee acting on behalf of a number of local committees). Present arrangements within the health care system in the UK for the handling of such data remain very unsatisfactory and put patient privacy at risk. PMID- 9724944 TI - Comorbidity of hospital-treated psychiatric and physical disorders with special reference to schizophrenia: a 28 year follow-up of the 1966 northern Finland general population birth cohort. AB - We studied the comorbidity of psychiatric and physical disorders in a sample (n = 11,017) from the unselected, general population, Northern Finland 1966 Birth Cohort. During the period 1982-1994, hospital-treated psychiatric patients were more likely than people without psychiatric diagnoses to have been treated for physical disease in hospital wards, 298 out of 387 (77%) vs 6687 out of 10,630 (62.9%) (OR = 2.0, 95% CI = 1.6-2.5). Injuries, poisonings and indefinite symptoms were a more common reason for hospital treatment in people with schizophrenia or other psychiatric disorder as compared with people without a psychiatric disorder. Men with psychiatric disorder had more than a 50-fold risk for poisoning by psychotropic drugs (OR = 52.6, 95% CI = 27.7-99.8), women with psychiatric disorder a 20-fold risk (OR = 19.0, 95% CI = 9.5-38.1) and schizophrenics more than a 30-fold (OR = 37.5, 95% CI = 19.1-73.8). Men with psychiatric disorders were more commonly hospitalised for a variety of gastrointestinal disorders and circulatory diseases (OR = 2.3, 95% CI = 1.2-4.4), as compared with men with no psychiatric disorder. Respiratory diseases (OR = 2.2, 95% CI = 1.2-4.2, vertebral column disorders (OR = 4.2, 95% CI = 1.8-9.9), gynaecological disorders (OR = 2.1, 95% CI = 1.2-3.6) and induced abortions (OR = 1.8, 95% CI = 1.2-2.7) were more prevalent in women with psychiatric disorder than in other women. Epilepsy was strongly associated with schizophrenia (OR = 11.1, 95% CI = 4.0-31.6). Nervous and sensory organ diseases in general (OR = 2.5, 95% CI = 1.1-5.8) and inflammatory diseases of the bowel (OR = 12.8, 95% CI = 3.8-42.7) were also overrepresented in schizophrenia when compared with people without a psychiatric disorder. Our results indicate that physicians must be alert for psychiatric disorder, and mental health professionals must be aware of the considerable morbidity in their patients. PMID- 9724945 TI - Mortality in men of Irish heritage in West Scotland. AB - Britons of Irish parentage have been found to exhibit poorer health and to die at a younger age than the general population. This paper expands the investigation of Irish mortality patterns in Britain, to include men with patrilineal Irish descent from the immigration of the 19th and 20th centuries. Five thousand, seven hundred and sixty-six male employees aged between 35 and 64 y were examined in 27 workplace settings in Glasgow, Grangemouth and Clydebank between 1970 and 1973. Twenty-one years' mortality follow-up was analysed from a survey involving a health questionnaire and medical examination, using name analysis to identify those of patrilineal Irish descent. Fitting Cox's proportional hazards model to date of death, using date of birth and Irish names as covariates, resulted in the patrilineal Irish showing elevated mortality from all cases (relative risk 1.22; 95% CI [1.08, 1.38]) and coronary heart disease (relative risk 1.53; 95% CI [1.27, 1.83]). Mortality risk for men with an Irish surname was also elevated for cerebrovascular disease (relative risk 1.30; 95% CI [0.86, 1.95]), respiratory disease (relative risk 1.17; 95% CI [0.73, 1.86]) and injury or poisoning (relative risk 1.42; 95% CI [0.78, 2.61]), although the low numbers of men dying from these causes, meant that differences did not reach statistical significance at the 5% level. No differences were observed for cancer or other causes. Previous work has shown high mortality for second generation Irish, whereas this study indicates high all-cause mortality and an excess of deaths for coronary heart disease in the much larger group of men with patrilineal Irish descent from the immigration of the 19th and 20th centuries. PMID- 9724946 TI - Risk factors for development of dehydration in children aged under five who have acute watery diarrhoea: a case-control study. AB - OBJECTIVE: To identify factors for development of dehydration in under five year olds with acute watery diarrhoea. DESIGN: Hospital based unmatched case-control study. SETTING: Diarrhoea Treatment Unit, Government Medical College Hospital, Nagpur, India. PARTICIPANTS: The study included 387 cases of diarrhoea having severe or moderate dehydration and 387 controls suffering from diarrhea with mild or no dehydration. RISK FACTORS: The study included infancy, female sex, religion, residing in urban slums or rural area, under nutrition, cessation of breast feeding during diarrhoeal episode, fluid intake decrease/stopped during diarrhoea, ORS not received, home available funds (HAF) not received, both ORS and HAF not received, non-washing of hands by mother before preparation of food, after defaecation, after disposal of faeces, history of measles in the previous six months, frequency of stools > 8/d, frequency of vomiting more than twice per day and temperature more than 99 degrees F, as risk factors for development of dehydration. STATISTICAL ANALYSIS: Univariate analysis included OR, 95% CI for OR and Chi-square test. Multivariate analysis was carried out by unconditional multiple logistic regression (MLR). RESULTS: This study identified the significance of infancy, religion, severe undernutrition, non-washing of hands by mother before preparation of food, frequency of stool > 8/d, frequency of vomiting > 2/d, history of measles in previous six months, withdrawal of breast feeding during diarrhoea, withdrawal of fluids during diarrhoea and not giving ORS, HAF or both during diarrhoea, in the outcome of development of moderate or severe dehydration. CONCLUSIONS: Timely intervention in the preventable risk factors included in this study may prevent the development of moderate or severe dehydration in the children suffering form acute watery diarrhoea. PMID- 9724947 TI - Developing guidelines for community child health staff and examining the referral pathways and outcomes of care in the support of emotionally and behaviourally disturbed children. AB - The East Sussex Public Health Commissioners had enquired whether referral guidelines were needed to ensure that the children with the greatest concerns achieved support from the Child and Adolescent Mental Health Services (CAMHS). Community child health doctors and nurses recorded all contacts with children with emotional and behavioural difficulties (EBD) over a one month period. Data on presenting difficulties and on intervention pathways were recorded. The same children were reviewed five months later and the outcome was measured by clinical assessment and by support offered. The work showed that child health staff were managing cases both through single advisory sessions and through planned on-going support. Additionally, the use of the three tier pathways recommended by the Department of Health was demonstrated. Analysis of recorded data was used to describe referral pathways and to give guidance on the characteristics of children who needed referral to CAMHS. PMID- 9724948 TI - Using radiology records to improve epidemiological information in paediatric fractures: a feasibility study. AB - OBJECTIVES: To assess the feasibility of using routine computerised radiology records for community injury surveillance data using fractures in the child population as an example. DESIGN: Radiology and in-patient computerised files were accessed to extract information concerning type of fracture, age, sex, and home address. Diagnostic coding of radiological report was carried out using the ICD-9 classification. Children were assigned to local authority wards using home postcodes derived from home addresses. Ward fracture rates were calculated using 1991 census data. The association between ward fracture rates and deprivation was explored using Townsend scores. SETTING: North Tyneside General Hospital. SUBJECTS: Children age 10-14 y receiving care as in-patients or out-patients for long-bone fractures. RESULTS: Between April 1991 and March 1996 a total of 497 long-bone fractures were identified. Fractures in boys exceeded those in girls by a ratio of 2:1. The most common fracture identified was of the radius and ulna. There was no evidence of an ecological association between long-bone fracture rates in children aged 10-14 y and social deprivation. CONCLUSIONS: Computerised radiological records may be used to improve epidemiological information concerning fractures. However, at present, considerable time and effort is required to access the information, to identify and to classify, long-bone fractures. Such data could be used to assist in the audit of clinical care and long-term outcomes, and to inform effective local planning and evaluation of injury prevention initiatives. PMID- 9724949 TI - The effect of a lindane and mercury polluting incident on the health of a community: the Somerton Health Survey. AB - OBJECTIVES: This study sought to identify possible illnesses of people exposed to lindane and methyl mercury following a pollution incidence in Somerton, Somerset, UK. METHODS: A self-administered questionnaire survey was posted to 1500 residents in three selected areas of Somerton to identify symptoms of possible illness over a 3 month period. RESULTS: There was a 74% response rate. People living near the stream had higher levels of reported mental symptoms and itching skin than in controls. Poisoning as the cause of the mental symptoms was excluded as the individuals had normal blood levels of lindane or mercury. Other symptoms were no higher in one area than another or from one time period to another. CONCLUSIONS: The survey, using controls in time and space, was able to explore, the pollution incident's contribution to the toxicity of residents and how this related to mental symptoms experienced by the residents. PMID- 9724950 TI - HBV serum marker detection and relative factor analysis of 2925 new students. AB - The HBV serum marker of 2925 new students was detected by enzyme linked immunosorbent assay. This produced results with a high degree of precision in a large study sample. The detection results showed that the HBsAg positive rate of new students is closely related to the family location, being either from urban districts or rural areas, and also the educational levels of their parents. PMID- 9724951 TI - Smoking patterns of university students in Eskisehir, Turkey. AB - Over the last decade, there has been a major decline in the prevalence of smoking among Western populations whilst in most developing countries there is a serious health problem of increasing smoking prevalence, especially among young age groups. The purpose of this study was to determine the prevalence of smoking among university students in Eskisehir in Turkey and to study the effects of some socio-demographic factors on the habit of smoking. Self-administered questionnaires were distributed to 1474 students (591 female and 883 male). This survey instrument contained a section soliciting socio-demographic information which was followed by questions relating to the students' smoking behaviour and the presence of a significant individual who has an effect on the smoking and drinking behaviour of the student. Multiple logistic regression was used to calculate odds ratios for the independent effects of the socio-demographic factors. The prevalence of smoking among university students was 42.5% (being highest among the students of the Faculty of Arts and lowest among the students of the medical faculty (60.9% vs 33.9%). The results of the analysis indicated that: being male, studying arts or education as opposed to medicine, being a final year student, being resident with friends, having a father with a lower education level and have a family member(s) who smokes were independently associated with smoking. The prevalence of smoking was greatest among those who drank alcohol (OR = 5.20). When comparing our results with reports from other countries, we conclude that the habit of cigarette smoking is one of the most important public health problems in Turkey. PMID- 9724952 TI - The impending kidney transplant crisis for the Asian population in the UK. AB - Kidney transplantation offers the opportunity of an improved quality of life for those patients suffering from renal failure. Unfortunately, this treatment is not available to all people as this is influenced by the increasing demand for a limited supply of suitable organs. This situation is particularly alarming for the UK's Asian population with their higher susceptibility to end-stage renal failure which has resulted in a greater demand for transplants. Consequently, the proportion of Asians on transplant waiting lists is growing rapidly. Coupled with this are problems of cross-racial tissue type matching which has led to longer waiting times for a transplant. The situation is clear, there is an urgent need to address the number of Asians requiring a kidney transplant otherwise the human and economic costs will be very severe. In the short term there needs to be a greater number of donors coming forward from the Asian communities to increase the pool of suitable organs. In the long term, there needs to be greater attention on preventive strategies to reduce the number of Asians requiring renal replacement therapy. PMID- 9724953 TI - The role of complementary medicine in European and Asian patients with inflammatory bowel disease. AB - Three hundred and eighty-two patients with known inflammatory bowel disease (IBD) (190 European and 192 Asians) and 190 with coeliac disease were sent a previously validated questionnaire to investigate patients' use of alternative medicine and their views on its effectiveness. Details sought included whether they have ever consulted an alternative practitioner, whether they had followed a course of treatment and its clinical effects. Information about where patients had heard about such alternative practitioners and whether they were told to discontinue their current allopathic medication was sought. Results were analysed after three consecutive mailings, including one in Gujurati to Asian patients. A randomly selected group was re-interviewed four months later. To validate the study alternative medicine practitioners were also interviewed to investigate what percentage of their attendees have IBD and how many of those clients were Asians. One hundred and fifty-eight questionnaires were returned from European patients with IBD (response rate = 83%), 145 from patients with coeliac disease (response rate = 76%) but only 81 Asian patients with IBD (response rate = 42%). Forty seven European and Asian patients with inflammatory bowel disease sought advice or treatment from an alternative practitioner, compared with only 11 with coeliac disease (chi(2) = 11.64, df = 12, P < 0.003). There was no significant difference in consultation rates between Asian and European patients with IBD (Yates corrected chi(2) = 0.78, ns). The most common practitioners consulted by all groups were homeopaths (n = 23) and herbalists (n = 27) but 20 patients consulted more than one practitioner at a time. Patients with coeliac disease and European patients with IBD had consulted osteopaths (n = 6) and reflexologists (n = 7). Ten patients with IBD had also attended a spiritualist and five Asian patients a hakim. Common sources of information about alternative remedies included friends and relatives (n = 13), the media (n = 11), word of mouth (n = 11) and family practitioners (n = 6). Most patients were advised to continue their current medications, although two had been told to stop and 10 advised to reduce the dose of their allopathic medications. Twenty alternative medicine practitioners stated that overall between 2-5% of their attendees have IBD with 10% of those clients being Asian. Asians preferred to consult Asian practitioners rather than European practitioners. There was no clear consensus as to whether complementary therapies were felt beneficial, although many patients with IBD believed them to be helpful. PMID- 9724954 TI - Internet and international health policy. PMID- 9724955 TI - Analysis of complex survey data--a solution using SAS. PMID- 9724956 TI - Promises made. PMID- 9724957 TI - Two new atypical antipsychotics: advantages and disadvantages. PMID- 9724958 TI - Ribozymes to the rescue: repairing genetically defective mRNAs. PMID- 9724959 TI - Propagating epigenetic states through meiosis: where Mendel's gene is more than a DNA moiety. PMID- 9724960 TI - Pathways to enlightenment. PMID- 9724961 TI - Population genetics and human origins--haplotypes are key! PMID- 9724962 TI - You are what you eat: a gene transfer ratchet could account for bacterial genes in eukaryotic nuclear genomes. AB - Recent phylogenetic analyses reveal that many eukaryotic nuclear genes whose prokaryotic ancestry can be pinned down are of bacterial origin. Among them are genes whose products function exclusively in cytosolic metabolism. The results are surprising: we had come to believe that the eukaryotic nuclear genome shares a most recent common ancestor with archaeal genomes, thus most of its gene should be 'archaeal' (loosely speaking). Some genes of bacterial origin were expected as the result of transfer from mitochondria, of course, but these were thought to be relatively few, and limited to producing proteins reimported into mitochondria. Here, I suggest that the presence of many bacterial genes with many kinds of functions should not be a surprise. The operation of a gene transfer ratchet would inevitably result in the replacement of nuclear genes of early eukaryotes by genes from the bacteria taken by them as food. PMID- 9724963 TI - Splitting the ATM: distinct repair and checkpoint defects in ataxia telangiectasia. AB - Ataxia-telangiectasia (A-T) is an autosomal recessive human disorder that, because of its multisystem nature, is of interest to scientists and clinicians from many disciplines. A-T patients have defects in the neurological and immune systems, telangiectasia in the eyes and face, and are, in addition, cancer-prone and radiation-sensitive. A-T cell lines have a range of diverse phenotypes including sensitivity to ionizing radiation and defects in cell-cycle checkpoint control. The ATM protein is a member of the PI 3-kinase-like superfamily, and it has been widely accepted that A-T cells represent mammalian cell-cycle checkpoint mutants and that the radiation sensitivity is a consequence of this defect. However, several lines of evidence suggest that A-T cells have distinct repair and checkpoint defects. A-T cells therefore appear to harbour dual checkpoint/repair defects. Here, we review the evidence supporting this contention and consider its implications for an analysis of the A-T phenotype. PMID- 9724964 TI - A locus control region regulates yeast recombination. AB - The yeast Saccharomyces can switch its mating type by a highly choreographed recombination event in which 'a' or 'alpha' sequences at the mating-type (MAT) locus are replaced by opposite mating-type sequences copied from one of two donors, HML and HMR, located near the two ends of the same chromosome III. MAT alpha cells 'know' to choose HML, while MAT alpha cells preferentially recombine with HMR. Donor preference is regulated by a 250 bp recombination enhancer, that controls recombination of the entire left arm of chromosome III. Recent studies have shown how this locus-control region is turned on and off. PMID- 9724965 TI - Understanding cell migration guidance: lessons from sex myoblast migration in C. elegans. AB - Studies of sex myoblast (SM) migration in the nematode Caenorhabditis elegans have shown that multiple guidance mechanisms cooperate to ensure the accurate and reproducible targeting of the SMs. Many issues arise in the analysis of SM migration, including the action of multiple guidance mechanisms, redundant sources of guidance information, the multiple uses of molecular components, and whether factors affect cell fate determination events or the guidance mechanisms themselves. These issues are common to many cell migration events and make the analysis of SM migration instructive to our general understanding of how cell migrations are controlled. PMID- 9724966 TI - Of genes and genomes and the origin of maize. AB - The crop plant maize (corn) is remarkably dissimilar to its recent wild ancestor, teosinte, making it an extremely interesting model for the study of evolution. Investigations into the evolution of maize are currently being performed at the molecular and morphological levels. Three independent lines of research are poised to shed light on the molecular basis of this spectacular transformation: (1) determining the structure and origin of the maize genome; (2) understanding the role of transposable elements in maize evolution; and (3) elucidating the genetic basis for morphological differences between maize and its wild ancestor teosinte. PMID- 9724968 TI - It's a knockout! PMID- 9724967 TI - Using metabolic pathway databases for functional annotation. PMID- 9724969 TI - Is HIV treatment becoming too complex for nonspecialists? PMID- 9724970 TI - Blind to the danger? PMID- 9724971 TI - Folic acid and the pill. PMID- 9724972 TI - Watch your step! PMID- 9724973 TI - Surgery in stereo. PMID- 9724974 TI - Surgery in stereo. PMID- 9724975 TI - The improving outcomes of coronary artery bypass graft surgery in Ontario, 1981 to 1995. AB - BACKGROUND: There is continuing uncertainty over the relative contribution of outcomes monitoring to changes in surgical outcomes over time. The authors studied temporal trends in the clinical characteristics and short-term outcomes of patients who underwent coronary artery bypass grafting (CABG) in Ontario before and after the implementation, in 1993, of a province-wide program to provide feedback on cardiac surgery outcomes. METHODS: The authors analysed data from hospital discharge abstracts on the clinical characteristics and in-hospital death rates of all 67,784 patients who underwent isolated CABG in Ontario between Apr. 1, 1981, and Mar. 31, 1996. RESULTS: Death rates were relatively stable during the first half of the 1980s, then declined gradually in the second half of the decade; this decline continued into the first half of the 1990s. In the 1990s patients were older than those in the 1980s, and a higher proportion had coexisting diseases. Between 1986/87 and 1995/96 the unadjusted death rate decreased by 52% (5.0% v. 2.4%) (p < 0.001). The annual relative rate of decline was approximately 6% (95% confidence interval 5% to 7%) in the period before the outcomes feedback program was implemented and about 9% (95% confidence interval 7% to 11%) in the period after implementation. INTERPRETATION: Rates of death after CABG have been declining steadily in Ontario since the mid-1980s. Outcomes based quality improvement interventions may facilitate; but are not a prerequisite for, improvements in the quality of surgical care. PMID- 9724976 TI - Kidney graft loss in children: implications for program development. AB - BACKGROUND: Graft survival in children who undergo kidney transplantation is lower than that in adults. The objective of the study was to review the experience of the first 22 years of operation of the regional pediatric kidney transplantation unit for Atlantic Canada, based at the IWK-Grace Health Centre, Halifax, and to use the results to improve graft survival. METHODS: All cases of kidney transplantation performed at the centre from 1971 to 1992 were reviewed and the data compiled with the use of a predetermined database outline. Data for first transplants were analysed and compared with those in North American databases. Of the 40 graft failures, 19 (48%) occurred within the first 3 months after transplantation, a rate similar to that at other centres. The overall survival rates tended to be slightly lower than those of international databases. The introduction of cyclosporine A as an immunosuppressant, in 1985, did not provide the expected marked improvement in survival. Infection frequently accompanied acute rejection, and there was a delay in treatment of infections and rejection after discharge home. On the basis of these preliminary findings, several program changes were made: 1) a sequential immunosuppression protocol was implemented, 2) the intensity of the medical surveillance was increased for the first 3 months after transplantation, with aggressive treatment of infections and rejections, 3) a dedicated pediatric transplantation team was established as a subset of the adult team and 4) pediatric-specific selection criteria for cadaver donors were formulated. After these changes were implemented, data were collected and analysed up to June 30, 1997. RESULTS: Graft survival rates at 1, 2 and 5 years improved dramatically. After the beginning of 1993, there were only 2 graft losses among 22 transplants. Only one of these occurred in the first 3 months, and it was due to recurrent disease. Twenty-four rejection episodes occurred (10 in the first 3 months after transplantation), but all were reversed easily with high-dose steroid therapy. INTERPRETATION: Sequential immunosuppression with close medical surveillance and early aggressive treatment of infection and rejection contribute to a marked improvement in kidney graft survival in children. PMID- 9724977 TI - Toward improved coronary artery revascularization: is this as good as it gets? PMID- 9724978 TI - Suicide and language. PMID- 9724979 TI - The one and only Mrs. Jones. PMID- 9724980 TI - The future of scientific medicine. PMID- 9724981 TI - The grammar of interpretive medicine. PMID- 9724982 TI - A light on medical practice in 19th-century Canada: the medical manuscripts of Dr. John Mackieson of Charlottetown. AB - During his long career as a physician in Charlottetown, Dr. John Mackieson (1795 1885) compiled 4 medical manuscripts: 2 sets of case records, a synopsis of the medical conditions that were common in his day and a formulary. As primary sources, these documents provide information about medicine in 19th-century Canada and augment our knowledge of the problems of medical practice in that era. They illustrate aspects of the work of Dr. Mackieson, a generalist with interests in surgery and obstetrics, and they facilitate an understanding of the rationale underlying the treatments that he and his contemporaries used. Although 150 years old, the case records can be appreciated for their relevance to the art of medicine. Two excerpts from the case records, presented in this article, provide a sense of Dr. Mackieson's writings and introduce a discussion on the significance of these manuscripts in relation to the ideas on disease and treatment that governed medical practice, both in Prince Edward Island and elsewhere in Canada, in the 19th century. PMID- 9724983 TI - I'm afraid it's bad news. PMID- 9724985 TI - US guidelines on way, but agreement on health impact of endocrine disrupters still lacking. AB - National and international agencies are looking at regulating certain chemicals that may have a detrimental effect on the endocrine system. But is there enough research to support such a move? PMID- 9724984 TI - Alcohol "on board," man overboard--boating fatalities in Canada. PMID- 9724987 TI - Emphasis on MD-patient communication to start in medical school, UBC decides. PMID- 9724986 TI - Armed forces worried as physicians flee from military life. PMID- 9724988 TI - Disability management efforts can reduce number of injuries, improve bottom line. PMID- 9724989 TI - Radical surgery for early gastric cancer. AB - The role of radical surgery for early gastric cancer has become a topic of considerable debate. Despite excellent results from Japan and several retrospective and uncontrolled trials, results from two large prospective randomized trials appear to demonstrate no benefit from D2 compared to the D1 resections. These trials have prompted a move away from radical lymph-node dissection. We argue that this reasoning is flawed and based not on the lack of efficacy of the D2 resection but in an attempt to reduce post-operative mortality and morbidity. Post-operative complications are largely a result of distal pancreatectomy and splenectomy and the relative inexperience of surgeons performing the operations. By preserving these organs and concentrating surgery to specialized centres the complication rate of radical surgery can be significantly reduced to approximate that of non-radical surgery. Lymph-node metastasis to the N2 nodes in early gastric cancer has been shown to be as high as 23%. Non-radical surgery poses significant risks of leaving residual disease. Radical surgery must remain the operation of choice if non-curative surgery for a curable condition is to be avoided. PMID- 9724990 TI - Justification for endoscopic treatment of subgroups of early gastric cancer. PMID- 9724991 TI - Campaign strategy and tactics for clinical trials in surgical oncology. AB - The strategic planning of the campaign against cancer and the employment of clinical trials on a national and supranational basis could be improved. Lessons and principles derived from other spheres of ordered activity might be usefully applied to the realm of clinical trials. Principles taught by military strategists may find practical applications in the development and prosecution of successful clinical trials of cancer therapy. In this paper we reflect upon the parallels and divergences between the principles of campaign strategy needed for the successful prosecution of military objectives and the efficient and effective prosecution of major clinical trials. PMID- 9724992 TI - False family history of breast cancer in the family cancer clinic. AB - AIMS: Awareness of hereditary breast and ovarian cancer in both the general public and the medical profession is increasing. Individuals who may be at risk on the basis of a family history are requesting risk determination and appropriate management in a variety of settings. Risk determination relies largely on pedigree analysis and epidemiological data. METHODS: We describe five individuals presenting in the family cancer or genetic counselling clinic where a factitious family or personal history led to erroneous risk estimation. Common factors in these families are a history of benign breast disease, poor communication within families, long survival with early onset or bilateral disease, a lack of detailed knowledge of the illness and treatment in close relatives and inconsistencies in the history in repeated consultations. PMID- 9724993 TI - Staging the axilla in breast cancer: an audit of lymph-node retrieval in one U.K. regional centre. AB - AIMS: Many surgeons undertake a level 1 axillary dissection in patients with invasive breast cancer. This dissection yields a variable number of lymph nodes for histological study. In this study, we report the consequences of this policy for staging of the axilla. METHODS: Between January 1995 and December 1995, 236 patients with a diagnosis of invasive breast cancer underwent axillary surgery. RESULTS: A median of eight nodes was identified (range 0-30). In only 11 patients less than four nodes were identified. An increase in the number of nodes harvested was associated with a higher proportion of node-positive patients and a higher number of metastatic nodes identified. CONCLUSIONS: We concluded that a standardized approach to axillary dissection consistently yields an adequate sample of lymph nodes for staging purposes. Most importantly, larger node samples yield higher detection rates for metastasis. This has a significant bearing on patient selection for adjuvant chemotherapy when compared with more limited sampling practices, including solitary sentinel node detection and biopsy. PMID- 9724994 TI - Total thyroidectomy for differentiated thyroid carcinoma: primary and secondary operations. AB - AIMS: There is considerable controversy concerning the most appropriate surgical treatment of patients with differentiated thyroid carcinoma (DTC). Although some authors have advocated subtotal thyroidectomy because of the decreased surgical morbidity and the lack of improved survival with a more extensive procedure, total thyroidectomy has been defended by others as a treatment of choice with lower morbidity. METHODS: We reviewed 106 consecutive patients who had been treated with total thyroidectomy for DTC to determine the complication rate. Forty-seven patients had primary operations and 59 had reoperations with completion of total thyroidectomy. RESULTS: Residual tumour in the remnant thyroid tissue was found in 53.8% of patients who underwent prophylactic completion thyroidectomy. Permanent hypoparathyroidism was present in one (0.9%) patient and accidental transient unilateral recurrent laryngeal nerve injury occurred in 2.8% of the entire series. No patient had permanent bilateral recurrent nerve palsy. Furthermore, the risk of complication was not significantly different when comparing primary total thyroidectomy or completion surgery. CONCLUSIONS: We recommend total thyroidectomy as a safe treatment for DTC with a low rate of morbidity. PMID- 9724995 TI - Vascular complications of isolated limb perfusion. AB - AIM: Isolated limb perfusion (ILP) is a complex vascular procedure which uses an extracorporeal circuit with high doses of cytostatic drugs and often hyperthermia for the treatment of extremity tumours. Our study investigated the incidence, treatment and subsequent outcome of vascular complications after ILP, about which little is known. METHODS: A retrospective study was performed, in which we found 10 vascular complications after 466 ILPs (2.1%). RESULTS: In eight patients, acute arterial obstruction developed in the immediate post-operative period, resulting from a thrombus at the arteriotomy site. Prompt reintervention with thrombectomy restored the circulation in all patients. One patient developed an arterial thrombus in the brachial artery due to compression of the surrounding tumour 12 days after ILP, which was successfully treated with thrombectomy and freeing the artery from the tumour. One patient was treated conservatively for digital micro-emboli. All complications occurred in women, maybe because of their generally smaller vessel size. No limbs were lost and all patients were free of any vascular problem after a median follow-up of 3.6 years. CONCLUSIONS: We conclude that vascular complications after ILP are rare, consist mainly of thrombosis at the arteriotomy site and can be successfully treated by prompt thrombectomy. Therefore, close observation of the peripheral circulation after ILP is necessary. PMID- 9724996 TI - Vascular interventions during post-chemotherapy retroperitoneal lymph-node dissection for metastatic testis cancer. AB - AIMS: Complete excision of nodal masses during post-chemotherapy retroperitoneal lymph-node dissection (RPLND) for metastatic non-seminomatous germ-cell tumours (NSGCT) of the testis often requires vascular surgical intervention. We report our experience of vascular interventions and complications in a large series of men undergoing postchemotherapy RPLND. METHODS: A retrospective review of vascular interventions during post-chemotherapy RPLND in 98 patients was undertaken (103 procedures). RESULTS: Macroscopic tumour clearance was complete in 95/98 men (97%). Vascular intervention was required in all cases. Major complications included acute tubular necrosis in one patient who had undergone left nephrectomy and extensive dissection around the right renal artery, progressive atrophy of the ipsilateral kidney in three men and a colonic stricture and associated colocutaneous fistula in one patient after division of the inferior mesenteric artery. Iliac and femoral venous thrombosis developed in both patients in whom the inferior vena cava (IVC) was excised and in one patient after partial IVC excision. Eight of the 98 patients have died. No late vascular complications have occurred to date. CONCLUSION: Complete tumour clearance can be achieved in most post-chemotherapy RPLNDs but invariably involves vascular intervention. Metastatic NSGCT should be treated by surgeons with the ability to undertake the vascular procedures required. PMID- 9724997 TI - Cervical lymph-node metastasis from cutaneous melanoma of the head and neck: a search for prognostic factors. AB - AIMS: To identify prognostic factors determining overall survival in patients with surgically treated neck node metastases of cutaneous melanoma. METHODS: A retrospective study was carried out in 70 patients who were surgically treated with curative intent for cervical lymph-node metastasis from cutaneous head and neck melanoma at our institution between 1960 and 1986. RESULTS: Median follow-up of the 14 patients still alive was 10 years. Of the 70 patients, 64 underwent a radical neck dissection, four a modified radical neck dissection and two a postero-lateral neck dissection. In 63 patients, the node dissection was for palpable involved nodes and in seven for microscopic disease. Survivals after 5 and 10 years were 23% (SE 5%) and 20% (SE 5%), respectively. Five-year survival was 62% (SE 17%) for patients with a melanoma less than 1.5 mm thick and 16% for lesions thicker than 1.5 mm. A regional recurrence in the neck occurred in 16 (23%) patients, of whom 14 were found also to have distant metastases. All patients with regional recurrence died from disease. CONCLUSIONS: Of the 15 patient-, tumour- and treatment-related factors tested, only the Breslow thickness of the primary lesion carried prognostic significance for survival (Bonferroni corrected P-value: 0.026). PMID- 9724998 TI - Primary malignant melanoma of the mucous membranes. AB - AIMS: To investigate malignant mucosal melanoma (MMM), a rare disease and one which has, till lately, remained unrecognized. Incidence of MMM ranges from 2 to 10% in various series. METHODS: We retrospectively reviewed the cases of malignant melanoma treated at the Regional Cancer Centre, Trivandrum, India, over a period of 15 years. RESULTS: A total of 163 cases of melanoma were identified, of which 21 had a lesion in mucosal sites. There were eight cases of upper aero digestive tract (UADT) melanoma, seven cases of rectal melanoma, five cases with lesions in the vagina and one case with a lesion in the urethra. The mean age of the patients was 52.8 years; mean age of presentation in urogenital and anorectal lesions was similar to overall mean age, while this was lower (47.5 years) for UADT lesions. Almost half of the patients presented with ulcer or nodule with or without pigmentation. Pain was present in three-quarters and vaginal bleeding was present in all cases of vaginal lesions. Almost one-third of the patients failed locally while another third developed distant metastasis during the follow-up period. A 2-year disease-free survival rate of 13.2% (95% CI: 2.2-34.1) was observed, which dropped to 6.6% (95% CI: 0.4-25.7) after 3 years. Survival appeared a little better in UADT melanoma compared to urogenital and anorectal melanoma; however, the difference was not statistically significant. CONCLUSIONS: Malignant mucosal melanoma seems to have an aggressive biological behaviour with a high incidence of local failure and metastasis. PMID- 9724999 TI - Anterior compartment resection of the thigh in soft-tissue sarcomas. AB - AIMS: Soft-tissue sarcomas of the anterior thigh present technical problems due to the proximity of the femoral vessels, and the disability caused by a standard anterior compartment resection. METHODS: We treated 44 consecutive patients with primary sarcomas in the anterior thigh with wide resection (n = 15), and modified (n = 26) or standard (n = 3) compartment resection. No patient had amputation as primary treatment. RESULTS: The overall rate of local recurrence was 6/44 (14%). Local recurrence was observed in 1/3 patients with standard anterior compartment resection and 5/41 (12%) of those with wide excision or modified compartment resection. It was noted in 1/6 (17%) patients with adjuvant radiation and 5/38 (13%) of those treated with surgery alone. One of six patients with local recurrence required amputation. The 5-year survival rate was 66% varying significantly according to grade. CONCLUSIONS: Limb preservation was possible in 98% of patients. Wide resection or modified compartment resection was feasible in the majority (93%) of patients resulting in improved function. PMID- 9725000 TI - Does the narrow operating field in perineal radical prostatectomy lead to more positive surgical margins? AB - AIMS: To assess the risk of leaving cancer-positive surgical margins in the perineal approach for radical prostatectomy as compared to the retropubic approach. METHODS: Seventy-six patients with clinically organ-confined prostate cancer (stage T1-2 NoMo) underwent radical prostatectomy. The 57 patients who underwent retropubic prostatectomy were compared to 19 patients in whom the perineal approach was undertaken. The two groups were compared for pre-operative PSA levels, clinical stage, biopsy Gleasson score, and any correlation between pre- and post-operative stage and grade of the disease and rate of cancer positive surgical margins. RESULTS: Although there were no significant differences in the rate of organ-confined diseases and specimen Gleasson score in the two groups, the rate of positive surgical margins in the perineal approach was significantly lower (15.7 vs 29.8%) and the rate of extracapsular disease with negative margins was significantly higher (15.7 vs 7%). CONCLUSIONS: The narrow surgical field in the perineal approach for radical prostatectomy does not pose a higher risk for positive surgical margins and it might be the procedure of choice in stage T1C prostate cancer with a Gleasson score of below 7. PMID- 9725001 TI - Technique of sentinel node biopsy in breast cancer. PMID- 9725002 TI - Gamma emission imaging in the management of breast disorders. AB - Breast cancer is the commonest malignancy to affect women. The malignant process may present clinicians with problems in establishing the diagnosis expeditiously, accurately staging the disease and assessing tumour response to primary systemic chemotherapy. Considerable recent interest has focused on the application of imaging techniques that utilize tumour-specific gamma-ray-emitting radiopharmaceuticals to resolve these problems. The wide availability of gamma camera systems makes single photon-imaging techniques, using radiopharmaceuticals incorporating conventional isotopes, attractive options. However, results concerning the detection of the primary breast cancer and the staging of axillary lymph nodes suggest that these techniques would appear to offer no significant advantages, when compared with those obtained using standard diagnostic methods. Dual gamma-ray-emission imaging by positron emission tomography (PET) may offer an alternative solution. Studies performed show that PET can accurately detect primary breast cancers, stage locoregional lymph nodes and visualize distant tumour metastases. Furthermore, PET may be able to monitor early tumour response to chemotherapy agents. It would appear, therefore, that dual gamma emission might have an important role to play in the management of patients with breast cancer. PMID- 9725003 TI - Breast carcinoma in residual breast tissue after prophylactic bilateral subcutaneous mastectomy. AB - We report a case of breast carcinoma 6 years after a prophylactic subcutaneous mastectomy. The incidence of breast carcinoma after prophylactic mastectomy is probably less than 2%. If removal of breast tissue is performed to prevent breast cancer, we advocate a complete simple bilateral mastectomy, including nipple areola complex, axillary tail and pectoral fascia. PMID- 9725004 TI - Association of breast cancer with meningioma. AB - We report a case of meningioma subsequently developed in a patient with bilateral breast carcinoma, which was originally thought to be single brain metastases. A brief review of the literature is presented with emphasis on the unique association between the two neoplasms, which suggests a possible hormonal relationship. The knowledge of this association is important in the differential diagnosis of patients with breast cancer who develop central nervous system manifestations. PMID- 9725006 TI - Giant splenomegaly caused by splenic metastases of melanoma. AB - Splenic metastases are rare and usually occur in the setting of widespread visceral metastases. Splenomegaly as manifestation of metastatic spread is extremely rare. A patient with melanoma and metastases to the skin and lung is described. He developed a giant painful splenomegaly. The splenectomy specimen demonstrated that the spleen was occupied by metastases from the melanoma. Metastases of melanoma may cause extreme enlargement of the spleen. If the patient's general condition is good, splenectomy is indicated in order to prevent spontaneous rupture of the spleen. PMID- 9725005 TI - Metastasis in a benign duodenal stromal tumour. AB - Gastro-intestinal stromal tumour (GIST) is increasingly recognized as a distinct entity within the group of soft tissue tumours. Mostly, GIST arises from the muscular components of the stromal layer, but the tumour may also originate from the autonomic nerve system, recently designated as gastro-intestinal autonomic nerve tumour (GANT). The majority of GIST is located in the stomach and small intestine; only 4% of GIST is found in the duodenum. Clinical and pathological criteria to differentiate benign from malignant GIST are not well established. Tumour size and mitotic activity are commonly considered as important features, predicting biological behaviour and outcome. It has been suggested that the clinical course of the GANT-type tumours may be more aggressive. We present a case of a radically resected duodenal stromal tumour with benign features, in a young woman, with metastases to the liver and peritoneum occurring 8 years after the initial diagnosis. PMID- 9725007 TI - Somatostatin-receptor-negative carcinoid tumour responsible for Cushing's syndrome. AB - A 30-year-old man presenting with Cushing's syndrome was admitted in 1995. A diagnosis of ectopically ACTH-secreting, primary-unknown abdominal carcinoid tumour was made. The patient's plasma ACTH and cortisol levels failed to decrease after short-term treatment with octreotide, and somatostatin-receptor scintigraphy did not show any accumulation in the tumour. The patient died 1 year after admission. This is a relatively rare somatostatin-receptor-negative case of this disease and it appears necessary to test for the presence of somatostatin receptors before treating with octreotide. PMID- 9725008 TI - The XIX Congress of the International Society for Analytical Cytology. Colorado Springs, 27 February to 5 March 1998. PMID- 9725009 TI - Determination of drug levels in human serum by circular dichroism. AB - A study of the applicability of circular dichroism (CD) for the determination of drug levels in human serum is described and a new method for the quantitative determination of optically active absorbing drugs having Cotton effects at wavelengths about 250 nm in human serum and/or plasma is proposed. The principal advantages of this method are speed, economy, and simplicity, no derivatization or chromatographic separation steps being needed. The validity of the CD determination was confirmed by analysis of variance, beta-lactam antibiotics being chosen as model drugs. In addition, the validation studies performed confirm the accuracy and precision of the proposed method. For beta-lactam antibiotics lacking Cotton effects above 250 nm, an alternative method based on the extraction of the drug from serum is considered. PMID- 9725010 TI - Unexpected difference in enantioselective retention on cellulase (CHB I) silica stationary phase caused by exchange of potassium for sodium ion in the mobile phase. AB - An increase in both retention and enantioselectivity for some beta-blocking agents was observed when exchanging potassium to sodium ion in the buffer used as mobile phase. A large effect of ionic strength on retention was observed, while the enantioselectivity was constant. PMID- 9725011 TI - Investigations of the metastable decay of DNA under ultraviolet matrix-assisted laser desorption/ionization conditions with post-source-decay analysis and hydrogen/deuterium exchange. AB - The fragmentation of positive ions of DNA under the conditions of matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) was investigated by post source decay (PSD) analysis and hydrogen/deuterium (H/D) exchange. Spectra of five different synthetic 4mer oligonucleotides were recorded. As a main result the hypothesis was confirmed that for these ions all fragment ions result from processes, initiated by protonation/deuteration of a suitable base followed by a loss of this base as a neutral or ion and further backbone cleavages. The three bases adenine, guanine, and cytosine all exhibit comparable lability for fragmentation. The spectra show evidence for an interaction of the adenine base with the phosphate backbone. Signals of fragments containing TT- and CT cycloadducts were observed in the spectra. PMID- 9725012 TI - Exploring infrared wavelength matrix-assisted laser desorption/ionization of proteins with delayed-extraction time-of-flight mass spectrometry. AB - We report a study of the application of delayed extraction (DE) to infrared wavelength matrix-assisted time-of-flight mass spectrometry (IR-MALDI-TOF-MS) of proteins. The shapes of the spectral peaks obtained with DE-IR-MALDI-MS are compared with those obtained from the same samples and matrix using continuous extraction (CE) IR-MALDI-MS. Application of DE results in significant improvements in the peak resolution, revealing spectral features (in proteins with molecular masses < 12 kDa) that were not resolved in the corresponding CE-IR Maldi mass spectra. Particularly significant is a series of peaks on the high mass side of the protonated protein peaks that arise through replacement of protons by adventitious sodium ions in the sample. We deduced that these sodium replacement species are a significant contributor to the broad tails (and resulting peak asymmetries) that are a feature of the DE-IR-MALDI mass spectra of proteins with molecular masses > or = 17 kDa. The peak width reduction observed in IR-MALDI by DE suggests that, as in UV-MALDI, the initial velocity distribution for ions produced in the MALDI process contributes to the peak broadness in the CE mass spectra. In a systematic comparison between DE UV-MALDI and DE IR-MALDI, we determined that photochemical matrix adduction is present in UV-MALDI but absent in IR-MALDI. In addition, we find that protein ions produced by IR irradiation are less internally excited (i.e., cooler), exhibiting less fragmentation, more Na+ replacement and/or unspecified noncovalent adduction, and more heme adduction with apomyoglobin. Thus, IR-MALDI appears to be a softer means for producing gas-phase protein ions than is UV-MALDI. It will be of considerable practical interest to determine whether large protein ions produced by IR-MALDI are sufficiently cool to survive transport through reflecting TOF mass spectrometers (without loss of small neutral species such as H2O, NH3, and CO2) and the extended time periods required for detection by quadrupole ion trap and Fourier transform ion cyclotron resonance mass analyzers. PMID- 9725014 TI - Investigation of low-voltage on-resonance ion selection for Fourier transform mass spectrometry. AB - Low-voltage on-resonance ion selection (LOIS) was recently introduced as an alternative technique for ion selection and storage. Under high pressure conditions and similar to the technique of quadrupolar axialization, unwanted (unselected) trapped ions are eliminated from the analysis cell through collisions with cell plates following orbital expansion. The ions remaining after tens of seconds of mass selection can be detected with better coherence, leading to improvements in ion detection and sensitivity. Here, experiments designed to test ion remeasurement and ion transfer capabilities are presented. Simulations of ion motion give insight into the possible mechanism of ion cooling, which does appear to be the same as that of the axialization process. Because of its ease of use, lack of need for additional hardware devices, and comparable ion selection results, LOIS is an attractive alternative for trapped ion experiments. PMID- 9725013 TI - Infrared, surface-assisted laser desorption ionization mass spectrometry on frozen aqueous solutions of proteins and peptides using suspensions of organic solids. AB - Surface-assisted, laser desorption ionization (SALDI) time-of-flight mass spectra of proteins and peptides have been obtained from bulk frozen aqueous solutions by adding solid organic powders to the solutions before freezing. Abundant analyte ions were obtained with a 3.28 microgram Nd:YAG/OPO laser. 20 compounds were evaluated as solid additives, and 16 yielded protein mass spectra. Successful solids included compounds like pyrene, aspartic acid, and polystyrene. The best results were obtained with nicotinic acid and indole-2-carboxylic acid, which yielded protein mass spectra anywhere on the sample and with every laser shot. Compared with ultraviolet-matrix-assisted laser desorption ionization on the same instrument, cryo-IR-SALDI had a comparable detection limit (approximately equal to 1 micro M), a lower mass resolution for peptides, and a higher mass resolution for large proteins. Approximately 2500 cryo-IR-SALDI mass spectra were obtained from a single spot on a 0.3-mm-thick frozen sample before the metal surface was reached. About 0.1 nL of frozen solution was desorbed per laser shot. The extent of protein charging varied between the SALDI solids used. With thymine, myoglobin charge states up to MH12(+12) were observed. It is tentatively concluded that observed ions are performed in the frozen sample. PMID- 9725015 TI - Derivatization of 5-fluorouracil with 4-bromomethyl-7-methoxycoumarin for determination by liquid chromatography-mass spectrometry. AB - A robust new analytical method has been developed for the determination of 5 fluorouracil (5-FU) in human plasma samples using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). The method is based on a liquid-liquid extraction procedure, precolumn derivatization, reversed-phase HPLC separation, and detection using atmospheric pressure chemical ionization and selected reaction monitoring. The derivatization agent used was 4-bromomethyl-7 methoxycoumarin. The internal standard for the assay procedure was a stable isotope labeled analog of 5-FU. The lower limit of quantitation was 1.0 ng/mL using 500 microL aliquots of plasma. Sample throughput on the mass spectrometer was approximately 17 samples/h (3.5 min/sample). The method was fully validated. The recovery of 5-FU averaged 76.1%. The accuracy of the assay, assessed from quality control samples, ranged from 99.1% to 104.3% (% theoretical). The overall interassay precision (% RSD) was 2.7%, and the intraassay precision (% RSD) ranged from 1.5% to 3.9%. The derivatized samples were found to be stable under sample analysis conditions and during refrigerator storage. The method was specific for the determination of 5-FU. PMID- 9725017 TI - Modified development in transgenic tobacco plants expressing a rolA::GUS translational fusion and subcellular localization of the fusion protein. AB - The rolA gene is transferred naturally by Agrobacterium rhizogenes to the genome of host plants, where it induces dramatic changes in development of transformed plants, including dwarfism and leaf wrinkling. The predicted translation product of the rolA gene is a small (11.4 kDa), basic (pI = 11.2) protein, which has no clearly significant similarity to sequences in the data bases. We have introduced into the tobacco genome a gene encoding a rolA::GUS fusion protein. Expression of this gene led to synthesis of an RNA and a protein of expected size, and the transformed plants exhibited the dwarfism and leaf wrinkling typical of rolA plants, but to a lesser degree than plants transformed with the wild-type rolA gene. The distribution of beta-glucuronidase (GUS) activity was compared in subcellular fractions of leaf extracts from plants expressing either the rolA::gus gene or a control gus construct. As expected, in the control plants, GUS activity was essentially cytosolic. In contrast, in plants expressing the rolA::gus gene the highest specific activity was associated with the plasmalemma fraction. PMID- 9725022 TI - Four classes of salicylate-induced tobacco genes. AB - We have identified and characterized fragments of 15 salicylic acid (SA) early response genes. The kinetics of induction and response to cycloheximide (CHX) treatment allowed classification of genes into four groups. Classes I-III are characterized by immediate-early responses, showing increased accumulation of mRNA within 30 min of SA treatment. Moreover, CHX did not block induction of these genes, indicating that latent cellular factors mediate the SA response. Class IV genes were induced more slowly, but still within 2 to 3 h of SA treatment, and required protein synthesis for expression. Although identified in this study as SA-responsive genes, several could also be induced by other compounds. Two genes were characterized in more detail, including isolation of cDNA sequences and additional analysis of gene expression. Sequence analysis revealed that the class I gene, C18-1, is the previously identified ethylene response element binding protein 1 (EREBP1), an ethylene-induced transcription factor for basic pathogenesis-related (PR) genes, whereas the class III gene, G8 1, is a novel sequence. G8-1 was found to be strongly induced only by SA and its active analogs and was exquisitely sensitive to low SA concentrations. These and other genes were found to be activated at early times following tobacco mosaic virus infection of resistant tobacco genotypes. PMID- 9725025 TI - Factors influencing the expression in vitro of Candida albicans stress mannoproteins reactive with salivary secretory IgA. AB - We have examined the influence of subinhibitory concentrations of several antifungals, the different glucose and ammonium sulphate concentrations in the culture medium as well as the strain variability on the expression in vitro of stress mannoproteins reactive with salivary sIgA in C. albicans and other Candida spp isolates. Irrespective of the conditions used, no reactivity with salivary sIgA was observed in yeast cells grown at 25 degrees C. However, when grown at 37 degrees C, all of the 10 C. albicans strains, but only 9 out of 28 non-C. albicans isolates studied showed reactivity with salivary sIgA. Cells grown at 37 degrees C in medium containing maximum concentrations of glucose and ammonium sulphate expressed the antigens reactive with sIgA during longer periods of time than the cells grown in medium with minimal concentrations of the same compounds. The regulatory role showed by the concentration of glucose and ammonium sulphate on the antigenic expression was subordinated, nevertheless, to the most important factor, the temperature of incubation. Only isolates showing low susceptibility expressed the antigens reactive with sIgA under the influence of subinhibitory concentration of antifungals. However, induced resistance to one of the antifungals tested (5 fluorocytosine) allowed the antigenic expression at elevated subinhibitory concentrations even in previous susceptible strains. In conclusion, in addition to the temperature, factors such as characteristics of the strain, the concentration of glucose and ammonium sulphate in the culture medium and the resistance to antifungals played a role on the expression of C. albicans antigens reactive with sIgA, which could be of clinical relevance in the course of infection. PMID- 9725026 TI - Yeasts in the gastrointestinal tract of preweaned calves and possible involvement of Candida glabrata in neonatal calf diarrhea. AB - To examine the possibility of a mycotic involvement in neonatal calf diarrhea (NCD) the presence of fungi was assessed in (a) the intestinal contents of dead calves and fecal samples submitted for routine laboratory examination, (b) fecal specimens, sampled once in winter and once in summer, of calves raised on 2 farms with different management systems, and (c) mucosal scrapings of various segments of the digestive tract of a diarrheic calf, massively shedding Candida glabrata. C. glabrata was the most prevalent fungal species isolated from the routine samples. It was the only fungus which was shed by the calves on the 2 farms, for continuous, more or less prolonged periods, but exclusively in the winter months. Diarrhea and C. glabrata shedding seemed to be associated. C. glabrata colonized the abomasum (the functional equivalent of the monogastric stomach) but not the other segments of the digestive tract of the euthanized calf. Based on the findings of this study it seems that while some yeast species may be considered as commensals of the digestive tract of calves, and consequently their isolation from intestinal contents or fecal samples has no clinical significance, others, such as C. glabrata may be involved in enteric pathogenic processes. Moreover, characteristics of the culture, previous chemotherapeutic treatments, the animal's age and possibly climatic conditions should be taken into account before deciding on the fungal isolate's clinical relevance. Determination of mycotic involvement in NCD by routine mycological examination of intestinal contents and fecal samples of diarrheic calves may be useful to avoid unnecessary and potentially harmful antibacterial therapy. PMID- 9725027 TI - Immunohistochemical detection for Actinomyces sp. in swine tonsillar abscess and granulomatous mastitis. AB - The tonsils of eleven pigs and the mammary glands of a sow were used to investigate actinomycotic lesions due to Actinomyces sp. infection. At necropsy, there was no abnormality on these tonsils, on the other hand, numerous abscesses containing sulfur granules were found in the mammary. Histopathologically, the Actinomyces sp. lesions were noted as crypt abscesses in the tonsils and as pus forming granulomas in the mammary glands. The microorganisms in both lesions were composed of bead-like cocci, bacillary cells and short, branching filaments, those cells being positive by the Gram's and Grocott's methods. Clubs were formed around the microbial clumps in these lesions. Immunohistochemically, there were cross-reactions between antibodies of Actinomyces sp. Chiba 101 (101) and swine actinomycetes of 7 species: A. bovis, A. hyovaginalis, A. israeli, A. naeslundii, A. pyogenes, A. suis) formerly Eubacterium suits) and A. viscosus. However it was possible to differentiate Actinomyces sp. 101 from them by absorption and dilution of the antiserum, then the microorganisms in the tonsillar crypt abscesses and the granulomatous mastitis were labelled with an immunoperoxidase technique using the absorbed Actinomyces sp. 101 antiserum. Thus, these immunolabelling properties are suggestive of the presence of 'A. suis' (Grasser) Franke 1973. PMID- 9725029 TI - Mycoflora and naturally occurring mycotoxins in poultry feeds in Argentina. AB - The purpose of this work was to determine the mycoflora and mycotoxins natural incidence in poultry feeds from 2 factories in Rio Cuarto, Cordoba. One hundred and thirty samples were taken from May/1996 to May/1997. The most dominant species isolated of poultry feed samples belonged to the genera Aspergillus spp 85% and Fusarium spp 70%. From Aspergillus genus eleven species were identified and A. flavus was the most frequent. Nine species were identified from the Fusarium genus and the predominant was F. moniliforme. Penicillium ranked third in the number of isolated cases. From this genus twelve species were collected of which P. brevicompactum (15%), P. restrictum (14%) and P. purpurogenum (12%) were the most common. The most significant mycotoxin from poultry feeds was aflatoxin B1 (AFB1) found in 48% of the samples, with levels ranging from 10 to 123 ng/g. For zearalenone (ZEA) the levels were 327 to 5, 850 ng/g and DON was not detected from the samples. Due to the fact that in Argentina there is little information about this topic, these data on poultry feeds in our region would be of worldwide interest. PMID- 9725031 TI - Shock chloramination: potential treatment for Chironomidae (Diptera) larvae nuisance abatement in water supply systems. AB - In the early 1990s, infestations of midge larvae (Chironomidae, Chironomus sp.) were discovered in the potable water system of Tel Aviv, Israel. Control measures, such as draining and cleaning tanks, spraying water into the tank's air space, and electrocution traps of midge adults, were either inadequate or ineffective. In this system, monochloramine concentrations of up to 0.75 mg/liter are used routinely as a secondary disinfectant. This chemical was tested in the laboratory as a toxicant of midge larvae. The mortality of 4th instar midge larvae after short exposure to high chloramine concentrations (LC50 values of 32 mg/liter for 75 min) suggested the efficacy of instituting a Shock Chloramination treatment program. Tanks were partially drained until they contained only 20 cm of water and were then temporarily disconnected. Chloramine was added to this water to produce a concentration of approximately 70 mg/liter for 1-2 h. Subsequently, all dead chironomids were flushed out, and the tank was refilled to attain the operational volume of water. A 2nd identical treatment of water in the tank was suggested 7 d later to kill midges from reproductive adults and egg masses that survived the 1st treatment. This treatment program was tested in commercial covered tanks and gave complete control of these pests for 6-10 wk. These results suggest that this treatment program may effectively prevent midge outbreaks in Israel's drinking water supply system during the height of the summer. PMID- 9725032 TI - The indole alkaloid tryptamine impairs reproduction in Drosophila melanogaster. AB - The plant-produced indole alkaloid tryptamine is one of a large array of neuroactive substances that may affect insect behavior, development, and physiology. We tested the role of tryptamine on insect reproduction using the fruit fly, Drosophila melanogaster (Meigen), as a model system. Measurements were made of reproductive success, oviposition rate, and preadult survival of insects on artificial diets containing tryptamine, its precursor tryptophan, as well as glycine and serotonin (5-hydroxytryptamine). Drosophila reproduction was reduced to 15% of controls when adult insects mated and the young were allowed to develop on medium containing 75 mM tryptamine. Tryptamine-induced depression in reproductive success was due to decreased oviposition rate and preadult survival. Serotonin, but not tryptophan or glycine, also reduced oviposition rate. Preference tests indicated that tryptamine may act as an antiattractant or antifeedant in this species. The accumulation of the indole alkaloid tryptamine in plants may provide a mechanism for reducing insect reproduction, which is potentially useful in protecting crop plants. PMID- 9725030 TI - In vivo effects of fumonisin B1-producing and fumonisin B1-nonproducing Fusarium moniliforme isolates are similar: fumonisins B2 and B3 cause hepato- and nephrotoxicity in rats. AB - Fumonisins are mycotoxins produced by Fusarium moniliforme, F. proliferatum, and related Fusarium species found on corn. They occur naturally in corn-based feeds and foods and are suspected human esophageal carcinogens. Fumonisin B1 (FB1), the most common homologue, causes the animal diseases associated with F. moniliforme. Hepato- and nephrotoxicities, disrupted sphingolipid metabolism, and liver cancer have been found in rats fed FB1. To determine the in vivo effects of diets containing fumonisins B2 (FB2) or B3 or (FB3), male rats were fed culture materials (CM) of FB1 non-producing F. moniliforme isolates to provide low (4.6 6.7 ppm), mid (32-49 ppm) or high (219-295 ppm) dietary levels of either FB2 (FB2CM) or FB3 (FB3CM). Other groups were fed culture material of an FB1 producing isolate (FB1CM) providing 6.9, 53 or 303 ppm total fumonisins (FB1: FB2: FB3 = 1.0: 0.38: 0.15) and a tenth group was fed a control diet having no detectable fumonisins. One-half (n = 5/group) the animals were killed after three weeks, at which time the toxicological and histopathological effects of the three culture materials were similar, mimicked the effects of FB1, and included decreased body weight gains, serum chemical indicators of hepatotoxicity, decreased kidney weights, and apoptosis of hepatocytes and kidney tubular epithelium. FB1CM, FB2CM, and FB3CM affected sphingolipids, causing increased sphinganine to sphingosine ratios (Sa/So) in both liver and kidneys. The remaining animals (n = 5/group0 were fed a control diet for three additional weeks. All body weight and tissue specific effects, including increased Sa/So, induced by the FB2Cm, FB3CM and low level FB1CM diets were absent following the recovery period. Except for mild biliary lesions found in the high dose of FB1CM group and a few apoptotic hepatocytes present in one mid- and two high-dose FB1CM rats, no evidence of toxicity remained in these groups, following the recovery period. PMID- 9725033 TI - Isolation of aerobic microbes from Ixodes scapularis (Acari: Ixodidae), the vector of Lyme disease in the eastern United States. AB - The spirochete Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Benner is transmitted by Ixodes scapularis Say, a vector of Lyme disease. As a 1st step into investigating the possibility of biocontrol of the tick, we identified the microbiota associated with the ticks. We collected, identified, and determined the sex of ticks from foliage and deer. Seventy-three initial bacterial isolates were recovered from 43 ticks (27 adults and 16 nymphs). The bacteria isolated from nymphs were qualitatively different (mainly gram-negative cocci) from the bacteria isolated from adult ticks (gram-negative and gram-positive rods). To determine long-term viability, these isolates were stored for 6 mo under laboratory conditions. After storage, 63 surviving bacterial isolates were characterized using the Biology System of identification by substrate utilization. Forty-four isolates were identified to the species level. Our characterization efforts focused on the 40 spore-forming bacteria, which could prove useful in the biocontrol of ticks. Eleven species of Bacillus were identified. Bacillus thuringiensis-B. cereus was the predominant species group isolated. Six isolates from this group formed crystals. PMID- 9725035 TI - Evaluation of three (Z)-9-tricosene formulations for control of Musca domestica (Diptera: Muscidae) in caged-layer poultry units. AB - A field trial comparing the effectiveness of toxic targets impregnated with different formulations of the Musca domestica L. female sex pheromone (Z)-9 tricosene was conducted in a caged-layer, deep-pit poultry unit in southern England. Targets baited with 5 g of technical grade (Z)-9-tricosene, or 5 g of a 40% polymer bead formulation, caught significantly greater numbers of M. domestica than control targets. This increase in attractiveness of the pheromone impregnated targets persisted for at least 24 wk. However, mean daily catch rates of M. domestica at targets baited with 5 g of a 2% wettable powder formulation did not significantly differ from control levels. Technical grade and bead formulations of the pheromone attracted significantly more males than females. However, the catches of female M. domestica at these pheromone-impregnated targets were significantly greater than female catches at control targets. Monitoring with sticky cards indicated that the introduction of toxic targets successfully suppressed adult M. domestica population density for up to 13 wk. Possible hypotheses explaining the effect of (Z)-9-tricosene on female attraction are discussed. PMID- 9725036 TI - [Experience as a model hospital for new good clinical practice (GCP) regulations]. AB - The new good clinical practice (GCP) regulations established by the Ministry of Health and welfare aim to promote clinical trials of new drugs that are ethical, reliable, and scientifically sound. According to these regulations, utmost importance should be placed on considerations of trial participant safety, welfare and the protection of his or her human rights. To this end, the responsibilities of the sponsor, the institution at which the trials are conducted, and the physician in charge have been clarified. These regulations stipulate that a system must be established which makes it possible for a third party to verify the course and conduct of a clinical trial. The new GCP regulations not only set standards for the development of new drugs, they also mark the beginning of a new era in medical science and treatment in Japan. PMID- 9725037 TI - [Carcinogenesis via microbial infection]. AB - Recent progress in the field of infectious diseases involving carcinogenesis has been striking. Extensive studies of Helicobacter pylori, and hepatitis type B and C virus showed that they are the primary cause of gastric cancer and hepatoma, respectively. Also some parasites such as Opistorchis viverrini and Schistosoma haematobium are also putative causes of cholangiocarcinoma and urinary bladder cancer, respectively. All of them require a chronic infection of more than 15 years. More than 50% of Japanese cancers are thus considered to be caused by chronic infection. The classic theory of carcinogenesis is radiation, chemicals and viral infection. Recent studies in free radical and biochemical research in our infectious diseases show all carcinogenesis involves free radical generation such as superoxide (O2.-), nitric oxide (NO), and their adducts peroxynitrite (ONOO-), H2O2 hydrooxyl radical (.OH), HClO, and NO2Cl as well as alkylperoxy radicals. All these molecular species are capable of modifying nucleic acid and DNA or RNA; furthermore a strand break is frequently observed, and hence potent mutagenicity and a probable cause of cancer. Thus, the unifying theory of carcinogenesis may most likely involve the mechanism of free radicals. This means a paradigm shift is needed in the public health policy for the tactics of cancer prevention. PMID- 9725038 TI - [Comparison of screening-detected and symptomatic lung cancer patients]. AB - Survival rates were studied in 416/27 screening-detected and 1,099/188 symptomatic patients with non-small cell lung cancer (NSCLC)/small cell lung cancer (SCLC). Screening-detected patients with both NSCLC and SCLC had earlier stage disease, showed better PS distribution, and a greater part of the patients underwent standard therapy than the symptomatic cohort with a significant difference. Median survival time, 5- and 10-year survival rates were 1,220 days, 44.4% and 34.9%, respectively, in the screening-detected patients, and 248 days, 11.3% and 7.5%, respectively, in the symptomatic patients with NSCLC. They were 584 days, 21.2% and 15.9%, respectively, in the screening-detected patients, and 257 days, 4.8% and 2.3% in the symptomatic patients with SCLC. PMID- 9725039 TI - [Characteristics of screening-detected breast cancer and trends in its therapy]. AB - Recently breast cancer has become one of the most common malignancies and one of the major causes of cancer death in Japan. In order to detect earlier stage breast cancer and to stop the increasing mortality, there is a greater need for mass screening. Mass screening performed by physical examination has a history of about thirty years in Japan, but it has had no effect in reducing breast cancer mortality. A new modality, mammography, is effective to detect non-palpable lesions, so it is expected to improve the accuracy of screening and reduce the mortality of breast cancer. Randomized trials in western countries have demonstrated that screening with mammography is effective for breast cancers in women aged 50 -69. In Japan, limited studies reported that effectiveness of screening was improved when mammography was combined with physical examination for detecting early breast cancer, but it is still unknown whether screening reduces breast cancer mortality. Since indications of breast conservation therapy have been rapidly extended it follows that breast conservation surgery for screening-detected cancer has increased. In Japan, because of its medical system, it seems impossible to conduct randomized trials for evaluating the efficacy of screening. Thus, it is important to interpret the screening data in western countries and to compare data with those from screening trials of Japan in the future. PMID- 9725040 TI - [Feature of screening-detected cancer and progress of treatment--esophageal cancer]. AB - The recent increase in the detection of esophageal mucosal cancer has been changing the direction of treatment. The rate of esophageal cancer detection in mass screening by X-ray is 0.008%, which is 1/13 that of gastric cancer. Moreover, the rate by endoscopy is higher; the former is 0.1% and the later is 0.6%. Further, endoscopic screening using iodine staining for a high risk group like alcoholism has 3.6% detectability on esophageal cancer and 1.7% on gastric cancer. The rate of cancer-detection of upper intestinal organs comes to 5.35% in all. Most of the esophageal cancer detected by endoscopy is mucosal cancer, which is treatable by endoscopic mucosal resection (EMR). The result of the treatment is 100% 5 year-survival in cases of m1 and 2 esophageal cancer. EMR of esophagus preserving treatment is truly effective for patients. Endoscopic examination using iodine staining for the high risk group is excellent for mass screening of esophageal cancer. PMID- 9725041 TI - [Characteristics and progress of treatment for gastric cancers detected by mass screening]. AB - We devised an X-ray examination to detect early gastric cancer, especially small gastric cancer, during the period between 1996 and 1997. As a result, the rate of gastric cancer detection by mass screening was 0.50%; and the rate of early gastric cancer detection was 79%. 1. Characteristics of gastric cancer detected by mass screening: Characteristics of gastric cancer were depressive-type undifferentiated cancer less than 4.0 cm diameter in work-place screening and depressive-type differentiated cancer less than 4.0 cm in area screening; 2. gastric cancer less than 2.0 cm in diameter was 75 (68%) of the 110 cases (total detected gastric cancer). 2) We have to consider that gastric cancer differs by sex and age. 3) EMR accounted for 39 (35%) of the 110 cases in treatment. This method resulted in a higher gastric cancer detection rate and served to maintain QOL. PMID- 9725042 TI - [Features of colorectal cancer with fecal-occult-blood tests--comparison with colorectal cancer with no screening]. AB - BACKGROUND: There is now evidence from three randomized controlled trials (from Minnesota, Nottingham and Funen) that screening average-risk individuals for colorectal cancer (CRC) with fecal-occult-blood tests (FOBT) can reduce mortality from CRC. In Japan, mortality rates from CRC have increased and mass screening with FOBT has been performed since 1992. Although there is growing proportion of CRC with FOBT, there is no conclusive evidence that they reduce mortality from this cause. We evaluated the feature of CRC with FOBT, as one of the methods for evaluation of the efficacy of screening with this test. METHODS: Between January, 1982 and December, 1996, 2071 cases with CRC resected in our hospital were considered. We evaluated the clinicopathological findings about age, sex, location of tumor, size depth, incidence of lymph node metastasis, stage and prognosis of CRC with FOBT (376) compared with CRC with no screening (controls; 1695). RESULTS: CRC with FOBT were earlier stage and smaller-sized cancers than controls. No significant difference was found in the incidence of lymph node metastasis at each depth and prognosis in each stage. CONCLUSIONS: By screening with FOBT, we can detect at an earlier stage and with a smaller cancer, with the same biological behavior as controls. Now, we must encourage colon screening, continuing to research ways to improve identification of high-risk subgroups and increase complicance. PMID- 9725043 TI - [Characteristics of uterine cancer detected by mass screening]. AB - In Miyagi Prefecture, mass screening for cervical cancer was initiated in 1961. We organized the project in cooperation with the Miyagi Cancer Society. Because cases detected in mass screening were treated earlier, their prognoses were better than cases diagnosed in outpatient clinics. A high proportion of patients detected in stage 0 and I showed improved prognoses. In 1983, the central government established the first Health and Medical Services Law for the Aged to support the project. The standardized death rate of cervical cancer in Miyagi Prefecture fell from 12.1 per 100,000 in 1961 to 4.0 per 100,000 in 1994. A case control study revealed that women who were screened, compared with women who had no prior screening, had an odds ratio for invasive cervical cancer of 0.14. The time interval of following the last negative smear was assessed, and we found that an odds ratio for a one-year interval was 0.09. However, there still remain problems, such as the lack of a further increase in the screening rate, fixation of examinees, and increase in the incidence of young women. Uterine body cancer is one of the increasing malignancies in Japan, as well as worldwide. Its epidemiological characteristics are as follows; 1) over 50 years old, 2) infertile and irregular menstruation, 3) post-menopause, and 4) atypical genital bleeding. Screening for uterine body cancer was started in 1987 under the second Health and Medical Services Law for the Aged. The target of the screening is limited to cases with the high-risk factors above described. We reported the results of mass screening in Miyagi Prefecture, elucidated the characteristics of uterine cancer detected by mass screening, and indicated the problems. PMID- 9725044 TI - [Characteristics of screening-detected prostate cancer on health checkup]. AB - Prostate cancer has the highest incidence and the second most common cause of death in U.S. men. In Japanese men, prostate cancer is not so common but is the most increasing neoplasm. Detection of early-stage cancer and early curative treatment are the most effective ways to reduce mortality from this disease. Therefore the mass screening system for prostate cancer to reduce mortality has become an important issue in recent years in Japan. The screening project for prostate cancer was conducted using only the prostate specific antigen (PSA) levels of primary screening with the cut off level of 4.0 ng/ml for candidates undergoing a medical checkup every 2 years. Totally, 20,338 men had health checkups every 2 years and 899 men requested the screening checkup for prostate cancer and their serum PSA level. Fifty (5.6%) men showed abnormal PSA level, 41 men had a second screening, 27 men underwent needle biopsy of prostate, and finally 14 cases of prostate cancer were detected. The detection rate calculated for prostate cancer was 1.56%. Thirteen cases out of 14 (92.9%) were clinically diagnosed as localized cancer. In 10 cases, total prostatectomy was performed and the pathological findings showed none of them to be clinically insignificant cancers. In conclusion, from our result the screening of prostate cancer using PSA only is considered to be very useful to detect early clinically significant prostate cancer. PMID- 9725045 TI - [Radiotherapy combined with daily administration of low-dose cisplatin for squamous cell carcinomas of the head and neck]. AB - The effects of radiotherapy combined with daily administration of low-dose cisplatin (CD DP) and radiotherapy alone for squamous cell carcinomas of the head and neck were compared clinically and histologically. There was no difference in the response rate between two groups with and without CDDP for pre-operative irradiation (30-40 Gy). However, the complete response rate in the radical irradiation group (60-70 Gy) with CDDP was significantly higher than without CDDP. In the histologic effect assessed by the classification of Shimozato in 9 of 19 patients undergoing radical irradiation with CDDP, 3 patients in this group showed a grade III effect, and the other 6 a grade IV effect. Only 5 of 11 patients having irradiation alone showed grade III or IV effect. In conclusion, full-dose radiotherapy combined with CDDP provided a high level of organ preservation and local control because of the high clinical and histological complete response rate at the primary site. PMID- 9725046 TI - [Combination chemotherapy with continuous infusion of low-dose cisplatin and UFT for advanced non-small cell lung cancer]. AB - A combination chemotherapy with continuous infusion of cisplatin (5 mg/body, day 1-5) and UFT (400-600 mg/body, day 1-5) was administered to thirteen patients for advanced non-small cell lung cancer. Myelosuppression and other toxicity were mild, and the quality of life of the patients was good. The response rate of thirteen patients was 23% (CR 0, PR 3). It was considered that chemotherapy using cisplatin (5 mg/body, day 1-5) and UFT (400-600 mg/body, day 1-5) was well tolerated and effective for the treatment of non-small cell lung cancer. PMID- 9725047 TI - [A randomized controlled trial with methotrexate (MTX), 5-fluorouracil (5-FU) and pirarubicin (THP) vs 5-FU alone in advanced or recurrent gastric carcinoma. Tokai Hokuriku THP Study Group]. AB - A randomized controlled trial was designed to investigate the therapeutic benefit of a combination chemotherapy consisting of MTX, 5-FU and THP in patients with advanced or recurrent gastric carcinoma. The patients were randomized into two groups; Group A patients (n = 37) underwent our combined chemotherapy, whereas Group B (n = 34) underwent chemotherapy with 5-FU alone as a control. There were no significant differences in various background factors between the groups. The median survival time was roughly 170 days after the randomization for the patients with advanced cancer (n = 26 for Group A and n = 25 for Group B), with no significant difference between the groups. Two long survivors, however, belonged to Group A. The median survival time of 161 days for Group A (n = 11) was longer than that of Group B (84 days, n = 9), but the difference was not statistically significant. The incidence of toxicities (leukopenia in particular) exceeding JCOG grade 3 was significantly higher for Group A, but no morbidity was observed. These results imply that patients with advanced or recurrent gastric carcinoma may benefit from a regimen of MTX, 5-FU and THP. PMID- 9725049 TI - [Antitumor effect of human lactoferrin against newly established human pancreatic cancer cell line SPA]. AB - This paper describes the antitumor effect of human lactoferrin against the human pancreatic cancer cell line SPA, which was newly established in our laboratory from a metastatic liver tumor of pancreatic origin. In tissue culture, the cancer cells proliferated rapidly at 16 hours of doubling time, and produced tumor markers into the culture medium at a high concentration. Subcutaneous and intraperitoneal transplantation of these neoplastic cells into nude mice resulted in tumor formation and carcinomatous peritonitis. The inhibitory effect of human lactoferrin on the cell growth of SPA was found both in vivo and in vitro. There was significant inhibition of cell growth in vivo at the concentration of 1 microgram/ml of hLf in the culture medium. And in the in vivo assay, hLf delayed the growth of subcutaneously transplanted tumors into BALB/c nude mice, and the effect was retained for two weeks. These results indicate the possibility that hLf will become one of the new drugs for adjuvant therapy against pancreatic cancer. PMID- 9725048 TI - [Correlation between pyrimidine nucleoside phosphorylase (PyNPase)/platelet derived endothelial cell growth factor and histological prognostic factor, and influences of 5'-deoxy-5-fluorouridine (5'-DFUR) administration on PyNPase levels. 5'-DFUR Joint Research Group in the Osaka Area for Gastric Cancer]. AB - PURPOSE: Pyrimidine nucleoside phosphorylase (PyNPase), among which thymidine phosphorylase (dThdPase) exists mainly in human tumor tissues, is an enzyme to convert 5'-deoxy-5-fluorouridine (5'-DFUR) to 5-fluorouracil. Recently, it was reported that dThdPase was identical to platelet-derived endothelial cell growth factor, angiogenetic factor. Therefore, we expect that there is possibility of dThdPase being a prognostic factor. METHODS: We investigated for a possible correlation between PyNPase activities in tumor tissues and prognostic factors of histological findings, examined the influences of preoperative oral 5'-DFUR administration to PyNPase levels and investigated for a correlation between HPLC methods and ELISA methods in patients with gastric cancer. RESULTS: Higher levels of PyNPase were observed in patients with advanced t,n,v, and ly factors. PyNPase levels decreased by 5'-DFUR in patients with differentiated cases. A high correlation was found between HPLC and ELISA methods. CONCLUSION: This study suggests that we must investigate possibility of PyNPase being a prognostic factor in more detail. PMID- 9725050 TI - [Steady state and disappearance of the metabolites of miproxifene phosphate in the treatment of breast cancer]. AB - We gave miproxifene phosphate to six patients with recurrent breast cancer and to one patient with advanced breast cancer. This drug was orally administered at a daily dose of 20 mg in the morning, and serial blood samples were obtained just before the drug administration. Treatment was discontinued in 16 days in the patient with advanced breast cancer. Tumor response was 2 PR and 4 NC (3MR) with an efficacy rate of 29%. Adverse effects of grade 2, such as anorexia, nausea or vomiting and fatigue with grade 3 flushing and chilling were observed in the one patient with advanced breast cancer. This climacteric syndrome disappeared after cessation of administration. In one of the patients with recurrent breast cancer, a calf muscle cramp was observed. Steady plasma levels were observed in one week or two for miproxifene and in 2 to 8 weeks for desmethyl miproxifene, which were active metabolites of miproxifene phosphate. The half lives of these metabolites for disappearance were calculated in three patients. That of miproxifene was 27 to 36 hours and that of desmethyl miproxifene was 156 to 202 hours. Miproxifene phosphate is a promising drug for breast cancer, and the results of pharmacokinetics of active metabolites will suggest the time to obtain maximum efficacy and for it to disappear. PMID- 9725051 TI - [Results of combination therapy of UFT-E with low-dose CDDP for patients with advanced recurrent breast cancer]. AB - CDDP, as a modulator for 5-FU, has already been described as a very effective treatment for gastrointestinal tract cancer. We administered a dose of 400 mg of UFT-E orally every day, and 10 mg of CDDP by drip infusion twice weekly, for more than 10 weeks to 12 outpatients with metastatic local, pulmonary, hepatic, osteal or multiple-organ cancer which showed a poor response to the pretreatment, and assessed its efficacy and drug toxicity. In terms of the clinical efficacy of this therapy, CR was noted in one patient and PR in 2 patients with a response rate of 25%. The incidence of drug toxicity was low. Complications included temporal transient nausea and anorexia in two patients and leukopenia grade 2 as bone marrow suppression in 3 patients. From the standpoint of QOL, as well as in terms of both antitumor effect and drug toxicity, the therapy mentioned above was believed to be effective for outpatients with advanced recurrent breast cancer. PMID- 9725053 TI - [Serum factors in cancer patients affecting the antitumor effect of 5 fluorouracil]. AB - A human colon cancer cell line, HCT-15, was found to proliferate in human sera. Sera of cancer patients undergoing continuous intravenous administration of 5 fluorouracil (5-FU) were tested for the antitumor effect by counting HCT-15 cells stained with Giemsa solution. 5-FU concentrations in those sera were not correlated with the antitumor effects. After HCT-15 cells were incubated in sera of cancer patients or healthy volunteers for 24 hours followed by removal of those sera, those cells were incubated in a culture medium containing 500 or 1000 ng/ml of 5-FU for 48 hours. It was shown that preincubation of HCT-15 cells with sera of cancer patients but not sera of healthy volunteers decreased the sensitivity of HCT-15 cells to 5-FU. These results suggest that there may be some factors in the serum of a cancer patient, which affect the antitumor effect of 5 FU. PMID- 9725052 TI - [The in vitro combination-effect of toremifene with CAF (cyclophosphamide, adriamycin, 5-fluorouracil) on growth of various human mammary carcinomas]. AB - Toremifene (TOR) is a new antiestrogenic agent, a triphenylethylene derivative that was developed as an alternative to tamoxifene (TAM). TOR has been observed to be more effective than TAM with milder toxicity at high doses. We examined the in vitro combination-effect of TOR with cyclophosphamide, adriamycin, 5 fluorouracil and three drug mixture (CAF) on the growth of various human mammary carcinomas. The combination shows a semi-additive or additive growth inhibitory effect on all estrogen positive cells used here except one cell line. In particular, the additive or synergic combination-effect was observed on TAM resistant cells. Furthermore, TOR exhibits a chemosensitizing activity in ADR resistant cells by expressing P-glycoprotein coded by MDR-1 (multidrug resistance gene). The chemosensitizing activity is dose-dependent of TOR. As described above, the combination of TOR with CAF shows more than a semi-additive effect in this experiment. In conclusion, the addition of high-dose TOR to CAF therapy might be useful for advanced/recurrent breast cancer. PMID- 9725054 TI - [Low-dose CDDP and continuous 5-FU treatment of advanced gastric cancer with peritoneal dissemination: a case with long disease-free survival]. AB - The patient was diagnosed to have gastric cancer (T3 N3 M0 P3, Stage IV b). We conducted LcFP therapy. CDDP, 7 mg/m2/day, day 1-5 i.v. drip for 2 hours, and 5 FU, 170 mg/m2/day, day 1-7, i.v. continuously for 24 hours. After 3 courses (one course: 4 LcFPs followed by one rest week), down staging (T3 N2 M0 P1. Stage IV a) and improvement of performance status were obtained, and then surgical resection was undertaken. After operation one course of LcFP therapy served as adjuvant chemotherapy. The patient has survived over one year and 8 months to date in a tumor-free condition. LcFP therapy promises to be useful in the clinical management of advanced gastric cancer. PMID- 9725055 TI - [A case of retroperitoneal malignant lymphoma successfully treated by chemotherapy]. AB - A 71-year-old woman was admitted to our hospital with the complaint of lower abdominal pain. CT scan revealed a huge retroperitoneal mass. On laparotomy the tumor was found to have extensively invaded the retroperitoneum, and an incisional biopsy was performed. Histopathological diagnosis of the biopsied specimen demonstrated diffuse malignant lymphoma of the pleomorphic type. Six cycles of CHOP chemotherapy were very effective for complete remission. Our review of 18 cases in the Japanese literature showed that chemotherapy and radiotherapy were effective for retroperitoneal malignant lymphoma, so due care must be taken in opting for surgery. PMID- 9725057 TI - [Prevention of cytomegalovirus pneumonia in patients with adult T-cell leukemia by nested PCR monitoring]. PMID- 9725056 TI - [Low-dose oral etoposide and subcutaneous injection of low-dose cytosine arabinoside (Et-A non i.v.) with or without minimum anthracycline for refractory acute non-lymphoblastic leukemia]. AB - Low-dose oral etoposide (50 mg for 10 to 14 days) and low-dose subcutaneous injection of cytosine arabinoside (15-20 mg/12 hours for 14 to 17 days) (Et-A non i.v.) were given to refractory acute non-lymphoblastic leukemia patients. Case 1 was a 47-year-old woman who had a relapsed M0. B-DOMP therapy failed but she recovered from severe infection. She had to go home to take care of her children. Et-A non i.v. was given for 14 days and 11 mg of mitoxantrone was added on the 3rd day. She achieved a complete remission (CR). Case 2 was a 72-year-old woman classified M4. Low-dose cytosine arabinoside (Ara-C) with additional daunorubicin (DNR) therapy failed. She became weaker due to the gastrointestinal toxicity of DNR. Et-A non i.v. was tried. Oral etoposide (Et) for 10 days and Ara-C for 14 days proved effective, and she achieved CR. Case 3 was a 59-year-old man with M4 developed from myelodysplastic syndrome, complicated with infection. Ara-C low dose therapy was begun, but it was not effective. It was switched to Et-A non i.v., 40 mg of DNR was added, with 12 mg of mitoxantrone and double administration of 3 mg of vindesine. He also achieved CR. Et-A non i.v. therapy is effective and available for induction therapy with low toxicity and convenient to daily life. PMID- 9725059 TI - [Staging classification of malignant lymphoma]. AB - Ann Arbor staging classification is used for staging of Hodgkin's disease. Recommendation by Cotswolds meeting is now widely accepted. For Non-Hodgkin's lymphoma, however, Ann Arbor staging classification can be available only for nodal lymphoma. Specified staging classifications will be suitable in extranodal Non-Hodgkin's lymphoma. PMID- 9725058 TI - [Peripheral blood stem cell transplantation in adult acute lymphoblastic leukemia as postremission therapy]. PMID- 9725060 TI - Blood transfusion and postoperative wound infection in intracapsular femoral neck fractures. AB - The relationship between blood transfusion and postoperative wound infection was studied in 695 operations for cervical hip fractures. A total of 156 (22%) patients were transfused with a total of 392 units of blood. A total of 31 (4.5%) patients developed a postoperative superficial or deep wound infection. Cultures identified Staphylococcus aureus as the cause in 71% and E. coli in 9.7% of the infections. A total of 11 out of 156 (7.05%) transfused patients developed a wound infection. In contrast only 20 out of 539 (3.71%) non-transfused patients developed a wound infection (p < 0.05). PMID- 9725061 TI - Knee injuries produced by recreational sports follow a different pattern than casual injuries. AB - Data was collected on 208 Chinese males who underwent knee arthroscopy following trauma in the period from January 1993 through December 1994. For the purpose of comparison and analysis, the patients were classified according to their sporting habits as sedentary patients, recreational athletes, or competitive athletes. There were no significant differences in age between the three groups (average age: 25.82 +/- 10.6; range: 12 to 73 years). A hemarthrosis was present in 122 patients, and an effusion in 35. The most common mechanism of injury was direct impact (105 patients), followed by a twisting injury (58 patients). The most common arthroscopic finding was an anterior cruciate ligament (ACL) tear (partial in-37 patients and complete in 43 patients). In both sedentary and athletic patients, a hemarthrosis was significantly associated with ACL and meniscal tears (p = 0.02). When comparing the pattern of arthroscopic pathology between sports and casual injuries, a significantly greater number of ACL tears (both complete and partial) were caused during sports activity (p = 0.032). Sports injuries also resulted in a significantly greater number of meniscal injuries (p = 0.028). However, sedentary patients suffered from a greater prevalence of tibial osteochondral fractures (10%) than did those in the sports group (5%) (p = 0.04). PMID- 9725062 TI - The results of total knee arthroplasty in workers' compensation patients. AB - Total knee arthroplasty was evaluated in 10 patients with post-traumatic osteoarthrosis secondary to work-related knee injuries (age- and sex-matched with 10 controls who had total knee arthroplasties for nonwork-related osteoarthrosis) to determine if Workers' Compensation status influenced treatment outcome. Using the Hospital for Special Surgery Knee Rating System (maximum possible score: 100), most recent follow-up scores averaged 64.1 for Workers' Compensation patients and 91.9 for controls. Subjective indices (pain, function) were significantly different between groups (p < 0.05), but objective indices (range of motion, strength, deformity, instability) were not. No significant differences were noted between groups on either immediate postoperative or most recent follow up radiographs (which were assessed for alignment and radiolucencies at implant surfaces, respectively). Suboptimal outcomes can be anticipated in total knee arthroplasties performed on Workers' Compensation patients, particularly in cases where claims have not been settled at the time of surgery. PMID- 9725063 TI - Long-term follow-up after intertrochanteric osteotomies for avascular necrosis of the femoral head. AB - Between 1978 to 1994, 110 osteotomies were performed in 94 patients. Fifty hips (40 patients) were included in this study. Six hips were in stage II avascular necrosis (Arlet-Ficat staging) and 44 hips were in stage III. In 14 patients an etiologic association was discovered, the remaining 26 were considered idiopathic. The postoperative assessment (Harris score) showed that there is no statistic differences between sexes. Results strongly depend on the amplitude of the necrotic (Kerboul) angle (p < 0.01), the preoperative mobility of the hip (p < 0.01), and age. The best postoperative results are usually obtained in young, active, patients with unilateral involvement, a necrotic angle of less than 200 degrees, a good preoperative range of hip motion, and in which the osteotomy is performed before the collapse of sequestrum. PMID- 9725064 TI - Analysis of rising from a chair after total knee arthroplasty. AB - The relationship between the range of motion following total knee arthroplasty (TKA) and the height of chairs when rising from a seated position was analyzed. Forty-six TKA subjects were evaluated; 16 had osteoarthritis, 30 had rheumatoid arthritis. Subjects were divided into two groups based on their degree of knee flexion (average: 96.0 degrees; range: 75 degrees to 135 degrees; group 1 contained 24 subjects with < 100 degrees of flexion and group 2 contained 22 subjects with > 100 degrees of flexion) in order to evaluate the acceptable knee flexion angle required for comfortably rising from a chair. To evaluate the flexion-extension angle of the knee and hip joints, three goniometers, a large reaction force plate, and a switch sensor on the chair surface to detect the initiation of rising were used. One chair had a height equal to the subjects' lower leg length, while the height of the second chair was 120% of the subjects' lower leg length. Analysis showed that TKA patients with less knee flexion (< 100 degrees) required a high angular velocity of the hip and excessive swing velocity to lift the trunk forward than did those with a larger degree of knee flexion (> 100 degrees). We concluded that a minimum of 100 degrees of postoperative flexion is desired and that a higher chair is more suitable for TKA patients. PMID- 9725065 TI - The suspected scaphoid fracture. How useful is a unit policy? AB - The records of 196 patients presenting with a clinical suspicion of scaphoid injury were reviewed to evaluate how junior accident and emergency doctors in a teaching hospital managed these patients. The management that was provided was assessed, and it was ascertained whether the presence of a unit policy meant that accident and emergency junior trainees managed patients accordingly. We found that 82% of patients were immobilized for 2 to 13 weeks, with 60 patients (37%) being immobilized for 6 weeks or more. Of the 196 patients presenting with clinical suspicion of scaphoid fracture, a definite scaphoid fracture was found in only 12%. Less than half of the patients (46%) were reviewed by senior accident and emergency doctors or by senior orthopaedic surgeons. Despite the presence of a unit policy, patients were being immobilized for prolonged periods in the absence of a radiographically evident scaphoid fracture. Advice from more experienced members of the staff was not being sought in dubious cases. PMID- 9725066 TI - Urografin injection demonstrated soft tissue ganglion communication with an intraosseous ganglion cyst. AB - Intraosseous ganglion cysts rarely occur in the hand or wrist. We present a case of a soft tissue ganglion cyst that communicated with the intraosseous ganglion of the scaphoid. Typical diagnostic studies include radiography, computed tomography, and magnetic resonance imaging. The diagnosis was made in this case by injection of Urografin into the soft tissue ganglion. The patient underwent excision of both ganglion cysts, resulting in a satisfactory outcome. PMID- 9725068 TI - Osteoid osteoma of the proximal phalanx of a toe. A case report. AB - A case of osteoid osteoma of proximal phalanx of second toe is presented. The clinical, radiological, and pathological features are discussed. Osteoid osteoma is rare in toe phalanges but should be considered when a patient presents with unexplained chronic pain in his or her toes. PMID- 9725067 TI - Spontaneous regression of a herniated disk. A case report with a four year follow up. AB - A 32-year-old male was seen with a "large" lumbar disk herniation on a magnetic resonance image. His pain was described as mild and he was treated nonoperatively. Four years later the disk showed almost complete spontaneous regression. No other reports of a longer follow-up MR image interval have been reported in the literature. PMID- 9725069 TI - Thoracic disk herniation in an 82-year-old patient. Treatment with the transpedicular approach. AB - A rare case of thoracic disk herniation in an 82-year-old female is reported. The patient was referred with a 2-month history of weakness and paresthesia of her left leg. On examination she had a severe paresis of the left leg and mild paresis of the right leg. Myelography and magnetic resonance imaging showed a T10 T11 disk herniation. The unilateral transpedicular approach was used and a large prolapse was surgically removed. The patient made an uneventful recovery and her neurologic function recovered to almost normal. PMID- 9725070 TI - Articular interposition of broken trochanteric wires. AB - A case of recurrent dislocation of total hip arthroplasty with intra-articular migration of broken trochanteric wires and interposition between the articulating surfaces of the prosthetic components is reported. The patient was treated with revision total arthroplasty with good result. This unusual complication should always be considered when performing trochanteric osteotomy in total hip replacement. PMID- 9725072 TI - Viruses and multiple sclerosis. AB - Animal models illustrate how viruses and host genetic factors may interact to cause immune-mediated demyelination. Similar mechanisms may take place in at least some forms of multiple sclerosis, a disease that is histopathologically heterogeneous. No 'multiple sclerosis virus' has been found yet, although recent data on human herpesvirus-6 antigens in multiple sclerosis brain warrant further investigation. Multiple sclerosis associated retrovirus, a recently described retroviral sequence isolated from multiple sclerosis material, is a member of the endogenous retrovirus-9 family. The association between the expression of this virus associated retrovirus and multiple sclerosis is only tentative. PMID- 9725073 TI - Recent advances in immunology in multiple sclerosis. AB - Multiple sclerosis is an inflammatory demyelinating disease of the central nervous system that is of possible autoimmune origin. This article is divided into three parts, reviewing recent advances in three selected topics regarding the immunology of multiple sclerosis. The first part addresses the consequences of T cell and oligodendrocyte death in the inflammatory lesions. The second covers the recent experimental evidence favouring the involvement of infectious agents in the pathophysiology of central nervous system autoimmune diseases. The third part concerns the mode of action of interferon-beta in multiple sclerosis. These new advances have lead to a better understanding of the immunopathology of multiple sclerosis and therefore open new therapeutic possibilities. PMID- 9725074 TI - Magnetic resonance imaging in multiple sclerosis. AB - Conventional magnetic resonance imaging (MRI) techniques acquire signal mainly from differences in relaxation properties and density of free water protons. Thus, the sensitivity in depicting lesions is high but pathological specificity is poor. Efforts are being made to increase the diagnostic power of MRI; better correlation with the clinical presentation and the use of better MRI criteria have increased the specificity of the conventional T2 sequences. New sequences, such as fast spin echo, turbo spin echo (TSE) or those derived from inversion recovery (fluid attenuated inversion recovery), have improved the detection of lesions. Acute phase monitoring focuses on changes in disease activity (new, recurring, enlarging, gadolinium-enhancing lesions) and chronic phase monitoring allows appreciation of the burden of the disease. However, MRI is considered as a secondary outcome measure in phase III trials because of insufficient correlation with the clinical disability. There is a continuing search for techniques that correlate better with clinical measures of the disease, such as the quantification of 'black holes' on T1-weighted images or the cerebral and spinal atrophy. The basic aspects of the pathological lesions in multiple sclerosis such as oedema, membrane disruption, demyelination, gliosis, cellular infiltration and axonal loss, can be studied more precisely by the new magnetic resonance techniques, which should better describe the actual clinical impact of the destructive process. In the past year the importance of axonal loss has simultaneously been confirmed by magnetic resonance spectroscopy and pathological findings. However, magnetization transfer imaging, magnetic resonance diffusion imaging and functional MRI are under intensive investigation for a better analysis of these different factors that impact on the reversibility of the patients disability. PMID- 9725075 TI - Symptomatic treatment in multiple sclerosis. AB - The increased awareness of the impact and complexity of management of symptoms in multiple sclerosis has resulted in advances in the understanding of their mechanisms, and in improvements in their measurement and management. It has also highlighted the paucity of evidence-based practice in this area and the need to develop agreed and comprehensive management strategies. PMID- 9725076 TI - Demyelinating inflammatory neuropathies, including Guillain-Barre syndrome. AB - The highly complex and multiple mechanisms responsible for the development of demyelinating neuropathies are reviewed, in particular Guillain-Barre syndrome and its variant Miller Fisher syndrome, chronic inflammatory demyelinating neuropathy, multifocal motor neuropathy, anti-myelin-associated glycoprotein neuropathy, as well as experimental models. Recent investigations into the role of auto antibodies against myelin proteins, or glycolipids have given insights into the pathogenesis of demyelinating inflammatory neuropathies. PMID- 9725077 TI - Functional brain networks in movement disorders. PMID- 9725078 TI - Neuroreceptor imaging: new developments in PET and SPECT imaging of neuroreceptor binding (including dopamine transporters, vesicle transporters and post synaptic receptor sites). AB - Positron emission tomography and single photon emission computed tomography have been used to measure receptor concentration and function through the use of a variety of radiotracers and data analysis techniques. Changes in presynaptic function and postsynaptic receptor concentration reflect both loss due to disease and compensatory responses. PMID- 9725079 TI - Experimental developments in movement disorders: update on proposed free radical mechanisms. AB - Free radicals have been implicated in the pathogenesis of movement disorders such as Parkinson's disease and Huntington's disease. Some basic aspects about free radicals as they relate to oxidative stress in neurodegeneration are summarized. Old and new experimental findings pertinent to oxidative damage in movement disorders are reviewed. Finally, the degree to which toxin-induced and genetically engineered experimental models have been useful in delineating parts of the mechanisms involved in the cascade of events that lead to neuronal death is emphasized. PMID- 9725080 TI - Fetal tissue transplantation [correction of transplanation] in Parkinson's disease. AB - Since the first successful attempts in 1990, human embryonic tissue transplantation has attracted the attention of multiple investigators and clinicians as a serious candidate therapy for Parkinson's disease. Although over two hundred patients have undergone the procedure, multiple issues and questions remain unresolved. We will address this topic emphasizing the recent advances in the technical aspects of the transplantation procedure in light of the limited animal and clinical experience available. PMID- 9725081 TI - Multiple system atrophy. AB - Although the precise definition of multiple system atrophy has been difficult, a recent consensus in diagnostic criteria for multiple system atrophy has been achieved. This should lead to progress in defining the underlying pathophysiology of the neuroendocrine, autonomic and motor deficits characteristic of multiple system atrophy. Hopefully, these developments will lead to effective treatment. PMID- 9725082 TI - Advances in Huntington's disease: implications for experimental therapeutics. AB - The gene mutation causing Huntington's disease was identified in 1993 as an expanded trinucleotide repeat within the coding region for a 348-kd protein called huntingtin. The mechanism by which this cytosine-adenosine-guanosine repeat produces the progressive signs and symptoms of Huntington's disease remains uncertain, but recent advances have begun to provide insights into this process. Promising developments include transgenic mouse models of Huntington's disease with neuronal intranuclear inclusions, the identification of differential neuronal features which might account for the selective vulnerability of neurons seen in Huntington's disease and further evidence for the role of excitotoxicity and impaired mitochondrial energy production. These observations have suggested new therapeutic strategies, and have lent further support for experimental therapeutics aimed at improving mitochondrial function and reducing excitotoxic injury. PMID- 9725083 TI - Dystonia. AB - Many different disorders have dystonia as the only or primary sign. The list of causes for dystonia increases yearly and now includes three mapped loci for primary torsion dystonia, although other susceptibility genes are suspected. Study of one of these primary torsion dystonia loci (DYT1) has culminated in the cloning of a gene which codes for a novel protein, torsin A. Physiological and positron emission tomography analyses suggest that dystonia results from impaired inhibition at cortical and subcortical levels; these physiological changes may in turn be due to striatal dysfunction and a mismatch or imbalance between the direct and indirect pathways. Future study of normal and mutant torsin A, as well as the identification of other primary torsion dystonia genes, should help elucidate the mechanisms underlying dystonia. PMID- 9725084 TI - Tic disorders: new developments in Tourette syndrome and related disorders. AB - The constellation of motor and vocal tics and certain of the other neuropsychiatric symptoms seen in Tourette syndrome are thought to have an organic basis, although the nature of the neurobiological lesion is uncertain. The syndrome is usually familial but the presumed genetic substrate has not been identified. A number of models currently under debate include a proposed autoimmune contribution. PMID- 9725086 TI - Movement disorders. PMID- 9725085 TI - Demyelinating diseases. PMID- 9725087 TI - Spondyloarthropathies. PMID- 9725088 TI - HLA-B27 and other predisposing factors in spondyloarthropathies. AB - Studies from around the world have confirmed the association between HLA-B27 and human spondyloarthropathies. The onset of many HLA-B27-linked arthritides follows an infection with enterobacteria. How bacteria interact with HLA-B27 and modify the immune system to give rise to the clinical disease is currently unclear. The roles of other genetic factors, including major histocompatibility complex class II genes and other genes located within this region (Tap/Lmp), have been postulated in certain spondyloarthropathies. We are using transgenic and knockout mice to answer some of these unsolved issues. This review discusses recent findings from our laboratories. PMID- 9725089 TI - Clinical, epidemiologic, and therapeutic aspects of ankylosing spondylitis. AB - Ankylosing spondylitis (AS) almost invariably starts before the age of 50, and clinical features suggestive of AS in older age should lead to consideration of other rheumatic disorders. Clinical manifestations of extraskeletal tissue such as renal amyloidosis and lung disease may occur. However, the detection of amyloidosis may not invariably infer poor prognosis, and associated lung disease may include apical fibrosis and also interstitial lung disease. Although the clinical significance and pathogenesis of osteoporosis in AS remain unclear, reduced bone mass may be found in a significant number of patients. Population surveys on AS have shown a correlation between the population frequency of HLA B27 and prevalence of AS. However, neither B27 subgroup distribution nor low frequency of B27 can explain the rarity of AS among certain African regions. Also representing an area of future research is the detection of both disease-related variables and sociomedical factors influencing the final outcome of this disease. PMID- 9725090 TI - Psoriatic arthritis. AB - Recent population-based studies have examined the incidence, prevalence, and survival rates of patients with psoriatic arthritis (PsA). Although there are still no completely satisfactory diagnostic criteria for PsA, Moll and Wright's criteria--although not a true classification/diagnostic scheme--are the most used. The sensitivity of these criteria is low (61%), as are the sensitivities of the European Spondyloarthropathy Study Group and Amor classification criteria for the whole spectrum of spondyloarthropathy. In some patients, PsA can occur only with peripheral enthesitis, particularly Achilles tendinitis or dactylitis. These patients may represent a subset of PsA that is not defined by the Moll and Wright or European. Spondyloarthropathy Study Group criteria and is, therefore, poorly recognized as such. Recent studies have analyzed the expression of adhesion molecules, cytokines, and chemokines in the synovial fluid and synovial membrane of patients with PsA. The therapeutic approach to PsA must be multidisciplinary, involving dermatologists and rheumatologists. Currently the most widely used second-line drugs are methotrexate, sulfasalazine, and cyclosporine. Used in combination, these drugs will probably become a well-established therapy for PsA. PMID- 9725091 TI - Undifferentiated arthritis and reactive arthritis. AB - The terms undifferentiated arthritis and undifferentiated characterize arthritides that do not fit into well-known clinical disease categories (e.g., seronegative rheumatoid arthritis and reactive arthritis) and that are an early stage or forme fruste of a definite rheumatic disease, an overlap syndrome between such diseases, or an unknown, etiologically undefined disease that remains to be differentiated from other types of arthritis or spondylarthritis. Undifferentiated arthritis and undifferentiated spondylarthritis share some clinical features with reactive arthritides. Recent data suggest that, at least in Chlamydia-induced reactive arthritis, the triggering bacteria persist in affected joints for some time during the course of the disease in a viable but nonreplicative state, providing an antigenic stimulus for a bacteria-specific immune reaction in the joint. The clinical manifestations of reactive arthritis include not only Reiter's syndrome or clinically suspected postinfectious arthritis but also undifferentiated oligoarthritis and spondylarthritis. The optimal treatment remains to be defined, but there is increasing data that antibiotic therapy is not as effective in cases of well-established reactive arthritis as has been suggested. PMID- 9725092 TI - Acute anterior uveitis and spondyloarthropathies. AB - Acute anterior uveitis (AAU) is characterized by sudden-onset, mostly unilateral exacerbations of an inflammation of the iris and ciliary body. The duration of illness is short if the patient is treated with corticosteroids. Half of all patients with any type of anterior uveitis are HLA-B27-positive, and more than half of the B27-positive patients have spondyloarthropathy. Ophthalmologists should therefore refer all patients with AAU who are HLA-B27-positive to a rheumatologist. Because attacks of AAU are extremely painful and frightening, most spondyloarthropathy patients with AAU will seek out an ophthalmologist on their own. The anterior chamber of the eye and the joints are mesenchymal cavities that are cleaned by macrophages. Anterior chamber-associated immune deviation is the mechanism by which specific regulatory T cells normally produce sufficient transforming growth factor-beta to impair inflammatory reactions that might hamper vision. Another mechanism of immune privilege is Fas-ligand induced apoptosis. Because the cells of the anterior eye express Fas-ligand, infiltrating cells are apoptotically killed. Comparable mechanisms may occur at a lower level in joints. The cause of AAU and spondyloarthropathy is unknown. B27 is probably only responsible for one quarter of the pathogenesis, other non-B27 genetic factors for another quarter, and unknown exogenous factors for the remaining half. It is possible that Gram-negative bacteria such as Klebsiella or Yersinia are involved in the pathogenesis in a yet unknown way. PMID- 9725093 TI - Arthritis and HLA-B27 in native North American tribes. AB - Whereas it is clear that HLA-B27 is increased in many North American natives, the prevalence varies a great deal. There is still a paucity of data on a large number of tribes. There is, however, a suggestion that HLA-B27 positive prevalence follows cultural/linguistic groupings. Reactive arthritis, sacroiliitis, and ankylosing spondylitis also appear to be increased in these people, but their relationship to HLA-B27 and possibly additional genetic predisposing factors is far from clear. Given these data and the known association of infectious agents, more extensive studies are warranted, especially in those tribes with a large enough population to achieve statistical significance. PMID- 9725094 TI - Bacterial arthritis. AB - Reports pertinent to bacterial arthritis in 1997 included two large, multi-year surveys of joint infection in patients from defined European health districts, noting trends including the declining incidence of gonococcal arthritis and an increasing number of prosthetic joint infections. Children with infected joints generally fare better than adults despite having proportionately more infections due to gram-negative organisms, of which Hemophilus influenzae comprises an ever smaller portion as the fastidious Kingella kingea is emerging. Joint infections remain an uncommon complication of immunodeficiency due to HIV, with responsible agents, affected sites, and clinical course also influenced by certain HIV comorbidities such as intravenous drug user and hemophilia. The rare immunodeficient patient with hypogammaglobulinemia retains a nearly unique susceptibility to joint infection with mycoplasmas, which can cause considerable morbidity if not promptly recognized and treated. Polymerase chain reaction can detect remnants of bacteria in the face of negative conventional cultures, but inoculation of synovial fluid into blood cultures bottles may be a more immediate and practical method to increase the yield in suspected septic arthritis. PMID- 9725095 TI - Mycobacterial and fungal arthritis. AB - Tuberculosis, atypical mycobacteria, and fungi are uncommon causes of infectious monarticular arthritis. The importance of these infections relates both to the difficulty in distinguishing them from other infectious and inflammatory arthritides as well as to their increasing incidence. Recently reported cases indicate more common use of newer molecular biological techniques such as polymerase chain reaction to achieve a more rapid and accurate diagnosis. PMID- 9725096 TI - Lyme disease. AB - A record number of cases of Lyme disease was reported in the United States in 1996. The steady increase in Lyme disease cases coincides with a growth in deer populations in endemic regions. Application of careful epidemiologic principles have led to the identification of two new pathogens that produce syndromes closely related to Lyme disease. Coinfection with Borrelia burgdorferi and the agent of human granulocytic ehrlichiosis have been documented. Coinfection with multiple organisms may alter the clinical course of Lyme disease. Better understanding of the pathogenesis of neuroborreliosis has provided clues into mechanisms responsible for persistent symptoms and will serve as a foundation for the design and implementation of appropriate therapeutic recommendations. A safe vaccine for the prevention of Lyme disease in humans has been developed, and preliminary reports from large clinical vaccine efficacy trials look promising. PMID- 9725097 TI - Retrovirus-associated rheumatic syndromes. AB - The influence of environmental factors in the initiation of autoimmune rheumatic diseases is still under debate. Infections with viruses (e.g., retroviruses) or expression of gene products encoded by endogenous retroviruses are believed to contribute to both loss of tolerance for autoantigens and immunosuppression. In various rheumatic disorders the detection of retroviral antibodies provides evidence for retroviral gene expression and suggests a role in the etiopathogenesis. Molecular mimicry, defects in apoptosis, altered autoantigens, and alterations of monocyte or macrophage as well as dendritic cell functions may contribute to the molecular mechanisms causing the loss of tolerance. On the other hand, destruction of lymphocytes during lytic retrovirus infections as well as envelope gene-env derived immunosuppressive retroviral gene products may prevent chronic inflammatory diseases and tissue destruction. Furthermore, retrovirus encoded super-antigens with the potency to skew the T-cell repertoire may seriously modify the host's immune system. PMID- 9725099 TI - Risk factors and prevention of fractures in the elderly. AB - Osteoporotic fractures represent a major public health problem in Western countries, and effective, preventive interventions are urgently needed. Whether it is worthwhile to begin treatment for osteoporosis prevention in elderly women has been the subject of some controversy. Although a 65- or 70-year-old woman is entering the highest period of risk for substaining hip and other fractures, it has been suggested that starting at this age may be too late for effective prevention. However, recent research supports the view that elderly women will greatly benefit from intervention such as physical training, dietary supplementation, or even estrogen replacement therapy. Several studies also suggest that focusing on the prevention of bone loss addresses only part of the problem. Preventing falls may also contribute substantially to reducing fracture rates in the elderly. PMID- 9725100 TI - Social and economic aspects of osteoporosis. AB - The first part of this article draws together descriptive studies on the direct medical costs of osteoporosis. The second part is devoted to economic evaluation of 1) osteoporosis treatment on the basis of clinical criteria and 2) preventive strategies, some of them including bone mineral density screening. These analyses point out the importance of taking into account quality of life and compliance issues. Research into dependency of the elderly has made it possible to obtain knowledge of the long-term economic consequences of osteoporosis. Development of research into dependency and its causes leads to the question as to whether the treatment and prevention of osteoporosis should be regulated in an isolated manner or whether it should be part of a larger health care policy with respect to aging. PMID- 9725102 TI - Pathophysiology and treatment of idiopathic hypercalciuria. AB - Nearly 50 years after its initial description by Dr. F. Albright, the term idiopathic hypercalciuria (IH) is still in use. The exact mechanism of hypercalciuria is still unknown despite extensive pathophysiologic investigations; recent advances represent the focus of this review. A precise definition of true IH is proposed, taking into account the various nutritional conditions influencing calcium excretion. The potential pathogenic mechanisms are discussed, and the limits of the classical Pak's pathophysiological classification are recalled. The evidence supporting the role of an increased intestinal calcium absorption, a defect in renal tubular calcium reabsorption, or an increased bone loss as a primary mechanism in IH are successively examined. Since overall available human data indicates that all three mechanisms may be found in IH, the hypothesis that a broader disorder encompassing all these various abnormalities may be involved in IH is discussed. Three global hypotheses to account for IH physiopathology are examined: a diffuse defect in fatty acid content of cell membranes, an increased expression of the vitamin D receptor of the 25(OH) vitamin D 1 alpha-hydroxylase, or of the calcium sensor receptor and a monocyte disease. Finally, the available clinical data justifying the therapeutic approaches are reviewed, and guidelines for dietary recommendations regarding calcium and also animal protein, sodium chloride, alcohol, carbohydrate, phosphate, and potassium intakes are proposed, and drug therapy indications are discussed. PMID- 9725103 TI - Treatment of skeletal involvement in neoplastic bone diseases. AB - Skeletal complications are frequent and increase morbidity dramatically in patients with malignancies. The role of bone resorption has been stressed leading to a key role for osteoclasts in the bone lesions. Specific and sensitive markers of bone resorption are now available. Bisphosphonates, powerful inhibitors of osteoclasts, can benefit patients with neoplastic bone diseases. PMID- 9725101 TI - Nutrition and osteoporosis. AB - Nutritional factors have a significant influence on the cause of osteoporosis. Calcium supplementation may be particularly effective in populations with a low calcium diet. Supplementations of 500 mg/d may produce about 4% gain in skeletal calcium in adolescents. Supplementations of 800 mg/d may prevent bone loss in postmenopausal women. The results of clinical trials also suggested that such supplementation may prevent hip and vertebral fractures in the elderly. The largest effect of calcium supplementation occurs in the first year of treatment, whereas sustained effects are not proven. Vitamin D supplementation may be particularly useful in vitamin D-deficient elderly. In this group, hip fractures may be prevented by vitamin D administration. Urinary sodium excretion is correlated with urinary calcium excretion in humans, and a direct effect of high sodium intake on loss at the hip has been demonstrated. Observational epidemiologic studies suggested a negative effect of a high protein intake on bone density, although there are no results from clinical trials to support this view. Dietary fiber, phytate, oxalate, and caffeine intake may have a small negative effect on calcium absorption. PMID- 9725104 TI - Spondylarthropathies. PMID- 9725105 TI - Infectious arthritis and immune dysfunction. PMID- 9725106 TI - Metabolic bone disease. PMID- 9725107 TI - A direct comparison of inhalant effects on locomotor activity and schedule controlled behavior in mice. AB - Increases in rates of rodent behavior have been commonly seen with exposure to abused vapors. In the 1st study, 30-min exposures to vapors of toluene, trichloroethane (TCE), or methoxyflurane produced increases in locomotor activity of mice at lower concentrations and decreases at higher concentrations. In the 2nd study, the effects of these vapors on schedule-controlled behavior were determined in mice lever pressing under a multiple fixed-ratio, 20-fixed interval (FI), 3-min schedule. Only concentration-related decreases in response rates were obtained in both components. In the 3rd study, toluene and TCE again produced only decreases in rates of responding under a simple FI 3-min schedule; biphasic effects were produced by methoxyflurane and amyl nitrite. The increases in rates of behavior often seen with abused vapors depend on the testing conditions. PMID- 9725108 TI - Associative tolerance to nicotine analgesia in the rat: tail-flick and hot-plate tests. AB - Previous assessments of associative nicotine tolerance may have confounded associative effects with novelty-induced stress effects, instrumental learning effects, or both. That is, subjects were tested in novel environments, allowed to practice the test response, or both during the tolerance development phase. In the first study, 32 male Sprague-Dawley rats were injected with various doses of nicotine and tested for nociception in the tail-flick and hot-plate tests to assess nicotine's analgesic effects. In the second study, 35 rats received nicotine explicitly paired or unpaired with a distinctive test context. All animals were equally preexposed to the test environment, and none had the opportunity to practice the test response. Paired rats developed greater nicotine tolerance than unpaired rats. This context-dependent (associative) tolerance effect was found with both tail-flick and hot-plate tests. PMID- 9725109 TI - Effects of amphetamine, butorphanol, and morphine pretreatment on the maintenance and reinstatement of cocaine-reinforced responding. AB - Three groups of rats (N = 23) self-administered intravenous cocaine (0.4 mg/kg/infusion) during the initial 2 hr of daily 7-hr sessions. During the remaining 5 hr of these sessions, responding produced saline infusions. On test days, each rat was pretreated with an assigned dose of either butorphanol, morphine, amphetamine, or saline before the self-administration session followed by a priming injection of cocaine (3.2 mg/kg i.v.) or saline at the beginning of the 4th hr. Results showed that both amphetamine (0.5, 1.0, and 2.0 mg/kg) and morphine (0.3, 1.0, and 2.0 mg/kg) pretreatment dose-dependently reduced cocaine self-administration whereas only amphetamine affected (increased) reinstatement responding. Butorphanol pretreatment (8 mg/kg) suppressed cocaine self administration and blocked the effect of the cocaine priming injections on pretreatment testing days. These findings indicate that (a) pretreatment drugs with similar stimulus properties potentiate reinstatement of responding and (b) pretreatment drugs can differentially affect maintenance and reinstatement of responding. PMID- 9725110 TI - State-dependent stimulus control: cuing attributes of acute cocaine rebound in rats. AB - Sprague-Dawley (Rattus norvegicus) rats were trained in a drug discrimination task using the state-dependent interoceptive stimulus attributes of cocaine's delayed or rebound effects (CDE) versus "normal" basal homeostasis. Rats were injected with either 32 mg/kg cocaine or equivalent volumes of saline (SAL), subcutaneously, 13 hr before the sessions. Rats demonstrated > 90% discriminative accuracy. Test sessions showed a time-dependent acute cocaine isodirectional rebound state that engendered a shift from predominantly SAL- to CDE-appropriate responding approximately 7 hr after the high training dose injection and lasted for approximately 10 hr (17 hr postinjection). The delayed or rebound state was dose dependent and engendered only a biphasic partial generalization with acute cocaine injections. There were no detectable levels of cocaine or any of its behaviorally active metabolites at the 13-hr postinjection interval. Tests conducted with various doses of lidocaine, chlordiazepoxide, N-methyl-d-aspartic acid, ketamine, and buspirone engendered SAL- or default-appropriate responding. The anxiogenic drug, pentylenetetrazole, produced partial generalization to the cocaine rebound cue. PMID- 9725111 TI - Attenuation of cocaine-induced locomotor activity by butyrylcholinesterase. AB - A primary enzyme for the metabolism of cocaine is butyrylcholinesterase (BChE). To determine whether the systemic administration of BChE can increase the metabolism of cocaine sufficiently to alter a behavioral effect, rats were tested in a locomotor activity chamber after receiving 17 mg of cocaine per kg intraperitoneally. In rats pretreated intravenously with 5,000 IU of horse serum derived BChE, the locomotor activity effect was significantly attenuated. BChE pretreatment increased plasma BChE levels approximately 400-fold. When added to rat plasma, this amount of BChE reduced the cocaine half-life from over 5 hr to less than 5 min. BChE altered the cocaine metabolic pattern such that the relatively nontoxic metabolite ecgonine methyl ester was produced, rather than benzoylecgonine. These results suggest that systemic administration of BChE can increase the metabolism of cocaine sufficiently to alter a behavioral effect of cocaine and thus should be investigated as a potential treatment for cocaine abuse. PMID- 9725112 TI - Memory association and personality as predictors of alcohol use: mediation and moderator effects. AB - We investigated the nature of the effects of memory associations on alcohol use and abuse. First, we determined if effects of memory associations on drinking problems are mediated entirely through the frequency of alcohol consumption or, alternatively, if such effects are more direct. Second, personality traits were assessed to evaluate whether they were confounded with memory association in their effects or whether they might moderate the effects of memory associations on alcohol use and abuse. The results showed that memory association measures directly and independently predicted alcohol consumption; these measures indirectly predicted problems from drinking, including drunk driving. None of the assessed personality variables moderated the predictive effects of memory association. The results are consistent with the view that memory associations influence behavior through cognitive processes that are not affected by personality traits or by cognitions emanating from such traits. PMID- 9725113 TI - Hyperbolic temporal discounting in social drinkers and problem drinkers. AB - Two studies compared participants, distinguished by their typical alcohol consumption, on the degree to which they discounted the value of delayed, hypothetical amounts of money. Heavy social drinkers in Study 1 and problem drinkers in Study 2 both showed greater temporal discounting than light social drinkers; this difference was stronger in Study 2. Both studies found that a hyperbolic function described temporal discounting more accurately than an exponential function. These results are consistent with extending behavioral theories of intertemporal choice to characterize the determinants of alcohol consumption. The discounting differences also are consistent with more general behavioral economic and economic theories of addiction, although the hyperbolic functional form is inconsistent with the exponential discounting function in economic theory. The drinker groups also differed on impulsiveness and time orientation questionnaires, with light drinkers being less impulsive and more future oriented; however, these measures were not strongly correlated with the measure of temporal discounting. PMID- 9725114 TI - Ecological momentary assessment in a behavioral drinking moderation training program. AB - We assessed predictors of self-reported excessive drinking (> 5 drinks) in a sample of heavy drinkers. Participants were randomly assigned to moderation training or a waiting-list control condition. They were trained in ecological momentary assessment (EMA) involving self-monitoring of drinking and other variables on a small hand-held computer, the electronic diary (ED). During the 8 week study, participants were compliant in their use of the ED for both random prompts and the entry of data related to specific drinking episodes. Generalized estimating equations were used to fit models involving predictors related to past history of drinking, aspects of the training program, drinking restraint, and episode-specific mood. The models indicated robust predictors of decreased and increased drinking. Our results suggest that EMA is a useful methodology for assessing drinking and related behaviors. PMID- 9725115 TI - Effect of stage of change on cue reactivity in continuing smokers. AB - Three classical conditioning models (the conditioned compensatory response, conditioned withdrawal, and conditioned appetitive motivational models) postulate that drug cues evoke physiological and emotional responses associated with motivational states that prompt drug use. There is accumulating evidence to suggest that factors other than classical conditioning can influence emotional and physiological reactivity to drug stimuli. This study tested whether stage of change affects the nature of reactivity to smoking cues among continuing smokers. Precontemplators (smokers not considering quitting) and contemplators (smokers considering quitting in the near future) watched videotapes containing smoking cues. Emotional and physiological responses to the smoking video were contrasted with responses to a neutral videotape. Precontemplators had lower heart rates than did contemplators in response to the smoking videotape. Both contemplators and precontemplators evinced increased positive affect in response to the smoking cue. A comparison sample of nonsmokers did not show any reactivity to the smoking cue. Implications of these findings for conditioning theories of smoking are discussed. PMID- 9725116 TI - Smoking urges affect language processing. AB - We examined the hypothesis that cigarette smoking urges take up working memory resources and interfere with language comprehension. Thirty-six smokers and 36 nonsmokers participated in the experiment. Eighteen members of each group were exposed to an imagery script that was intended to elicit a smoking urge in smokers, and the other 18 were exposed to a neutral script. The participants then performed a sentence comprehension task. The results showed that the urge script had a detrimental effect on the smokers' comprehension accuracy. As expected, there was no urge effect for the nonsmokers. Furthermore, smokers read the sentences significantly faster than nonsmokers. The results are discussed with reference to cognitive theories of urges and a capacity model of language processing. PMID- 9725117 TI - Nicotine-mecamylamine treatment for smoking cessation: the role of pre-cessation therapy. AB - The nicotinic antagonist mecamylamine was evaluated in a randomized smoking cessation trial. Four groups of participants (n = 20 per group) received nicotine plus mecamylamine, nicotine alone, mecamylamine alone, or no drug for 4 weeks before cessation. After the quit-smoking date, all subjects received nicotine plus mecamylamine treatment for 6 weeks. Nicotine skin patches (21 mg/24 hr) and mecamylamine capsules (2.5-5.0 mg twice per day) were used. Precessation mecamylamine significantly prolonged the duration of continuous smoking abstinence; abstinence rates at the end of treatment were 47.5% with mecamylamine and 27.5% without mecamylamine. Nicotine + mecamylamine reduced ad lib smoking, smoking satisfaction, and craving more than either drug alone. Moreover, the orthostatic decrease in blood pressure caused by mecamylamine was offset by nicotine. Mecamylamine before smoking cessation may be an effective adjunct to nicotine patch therapy. PMID- 9725119 TI - A new open-tubular approach to capillary electrochromatography. AB - While most capillary electrochromatography is based on the use of packing materials similar to those for HPLC, this method creates a new surface on the inner wall of the fused-silica tubing that is then modified with various organic moieties. In this format there is no need for frits, and bubble formation is not a serious problem. With 50-microns-i.d. capillaries, molecules are separated by a combination of differences in electrophoretic mobility and solute/bonded phase interactions. A number of examples are presented, which include tetracyclines, peptides, and proteins. Peaks for all compounds, even basic ones, exhibit both good efficiency and symmetry. The reproducibility of migration times is excellent even after extended use of the column. PMID- 9725118 TI - Advances in capillary electrochromatography. AB - This paper presents practical aspects of capillary electrochromatography (CEC). Preparation of capillary columns, including terminating frits and immobilization of the packed bed, is described in detail. Longevity and reproducibility of CEC columns is demonstrated. Instrumental aspects, sample introduction, sensitivity of detection, and gradient elution are discussed in detail. A number of examples that illustrate the versatility of CEC, in particular, for providing an isocratic separation alternative to conventional gradient separation in HPLC, are given. PMID- 9725120 TI - Rapid separation and quantification of major caseins and whey proteins of bovine milk by capillary electrophoresis. AB - A rapid capillary zone electrophoresis (CZE) method was established for separating and quantifying major casein and whey proteins in milk. Optimum sample preparation and electrophoretic conditions in a coated capillary maintained at 40 degrees C allowed accurate and reproducible quantification of milk proteins in a single analysis. Sample and run buffer allowed caseins to be maintained in solution by using a combination of urea and a nonionic detergent in phosphate buffer at pH 2.5. Quantitative CZE protein data were derived by calculating percentages and concentrations (mg/mL) of alpha-casein, beta-casein, alpha lactalbumin, and beta-lactoglobulin. Calibration curves followed linear relationships with highly significant (p < 0.1) correlation coefficients. Relative standard deviations of less than 0.82 (%) for migration times and 2.18 (%) for percent protein indicated that the technique was reproducible. Electrophoretic protein profiles of fresh bovine milk and rehydrated dry milk showed marked quantitative differences in whey protein concentrations. Whey protein represented 12.37 +/- 0.07% beta-lactoglobulin and 3.05 +/- 0.08% alpha lactalbumin of total protein in typical fresh milk, while only 1.90 +/- 0.16% beta-lactoglobulin and 0.86 +/- 0.04% alpha-lactalbumin of total protein were detected in a commercial rehydrated milk powder. By quantifying these differences, the established technique may allow the detection of substitution of fresh milk with rehydrated milk powder. The accuracy and reproducibility of the technique permitted the quantitation of individual protein concentrations in milk samples, which agreed with ranges reported in the literature. CZE may be well suited for routine use by dairies and regulatory agencies, since it allows the determination of milk proteins in less than 60 min. PMID- 9725121 TI - Capillary electrophoretic mobility shift assay (CEMSA) of a protein-DNA complex. AB - The utility of capillary electrophoresis in the study of DNA-protein binding is demonstrated, using the minimal DNA binding domain of the onco-protein c-Myb (R2R3) and a specific target DNA sequence as a model system. The capillary electrophoresis method is based on simple UV detection at 260 nm with a linear polymer buffer and a coated capillary, and requires no labeling or derivatization of the DNA. A specific protein-DNA complex is observable as a retarded peak, which increases with increasing protein concentration with a corresponding reduction in the free DNA peak. With DNA and protein preparations of known concentrations, a test for sequence-specific binding can be completed within 10 min. PMID- 9725122 TI - Stacking of neutral analytes in micellar electrokinetic chromatography. AB - On-line concentration techniques for neutral analytes by sample stacking in micellar electrokinetic chromatography are reviewed. Discussions regarding the fundamentals and practical applications are conveyed. A high gain in sensitivity of 10- to more than 100-fold using normal capillary cell dimension is provided without crucial loss of resolution by the techniques. More than 1000-fold gain in sensitivity (lowering limits of detection to the nM range) is obtained together with an extended pathlength cell. PMID- 9725123 TI - Prediction of selectivity for the separation of double-stranded DNA fragments in electrophoresis. AB - The electrophoretic resolution (selectivity) of double-stranded DNA (dsDNA) restriction fragments is studied for different organisms (phi X174, lambda phage, and pGEM-3 plasmids) by various separation and detection methods. The selectivity can be predicted by plotting the difference in chain length over their average chain length (base pair difference/average base pair magnitude of delta bp/Ave. bp) against each DNA fragment pair. Experimental conditions and the nature of the DNA can influence the predicted results. PMID- 9725124 TI - Determination of suramin by micellar electrokinetic chromatography with direct serum injection. AB - A simple and reproducible micellar electrokinetic chromatography (MEKC) method has been developed for the quantitation of suramin serum levels to be used for its therapeutic drug monitoring (TDM). A running buffer solution of 20 mM sodium borate, 75 mM sodium dodecyl sulfate (SDS), and 4 M urea at pH 9.2 were employed and samples were introduced directly into the capillary. The voltage applied for sample separation was 25 kV and UV detection was at 254 nm. Linearity was proved over the range 47.6 micrograms/mL-523.6 micrograms/mL of suramin (r: 0.9996). Orange G was used as internal standard and no interferences of common drugs simultaneously administered to patients were observed. Recovery values of the intraday and interday assays were between 92.5 and 97.2%. PMID- 9725126 TI - Proposition: radiation hormesis should be elevated to a position of scientific respectability. PMID- 9725125 TI - Real-time B-mode ultrasound quality control test procedures. Report of AAPM Ultrasound Task Group No. 1. PMID- 9725127 TI - Calculation of enhanced dynamic wedge factors for symmetric and asymmetric photon fields. AB - A method is introduced to calculate wedge factors for an enhanced dynamic wedge (EDW). An analytic formula has been derived that allows the determination of wedge factors at the center of symmetric and asymmetric photon fields. The formalism is an extension of the "MU fraction approximation," which holds that the dynamic wedge factor is equal to the fraction of MU delivered to the point of calculation. Extensive data are presented, comparing measured enhanced dynamic wedge factors with the current method and the MU fraction model for both symmetric and asymmetric fields. For both 6 and 18 MV photons, the current method demonstrates improved results: Agreement to within 1% is obtained in all symmetric fields and within 2% for all asymmetric fields compared with discrepancies of up to 4% obtained with the MU fraction model. PMID- 9725128 TI - Verification of the omni wedge technique. AB - The optimal field shape achieved using a multileaf collimator (MLC) often requires collimator rotation to minimize the adverse effects of the scalloped dose distribution the leaf steps produce. However, treatment machines are designed to deliver wedged fields parallel or perpendicular to the direction of the leaves. An analysis of cases from our clinic showed that for 25% of the wedged fields used to treat brain and lung tumors, the wedge direction and optimal MLC orientation differed by 20 degrees or more. The recently published omni wedge technique provides the capability of producing a wedged field with orientation independent of the orientation of the collimator. This paper presents a comparison of the three-dimensional (3D) dose distributions of the omni wedged field with distributions of wedged fields produced using both the universal and dynamic wedge techniques. All measurements were performed using film dosimetry techniques. The omni wedge generated fields closely matched the conventional wedged fields. Throughout 95% of the irradiated volume (excluding the penubra), the dose distribution of the omni wedged field ranged from +5.5 to -3.5 +/- 1.5% of that of the conventionally wedged fields. Calculation of the omni wedged field is as accurate as conventional wedged field calculation when using a 3D treatment planning systems. For two-dimensional treatment planning systems, where one must assume that the omni wedged field is identical to a conventional field, the calculated field and the delivered field differs by a small amount. PMID- 9725129 TI - Multileaf collimator leaf sequencing algorithm for intensity modulated beams with multiple static segments. AB - The "stop and shoot" method of producing intensity modulation using combinations of static multileaf collimator (MLC) segments has a number of advantages including precise dose delivery, easy verification, and general availability. However, due to the potential limitation of prolonged treatment time, it is essential to keep the number of required segments to a reasonable number. We propose an algorithm to minimize the number of segments for an intensity modulated field. In this algorithm, the sequence of delivery intensity is proposed to be a series of powers of 2, depending on the maximum intensity level in the matrix. The MLC leaf position sequence is designed directly on the two dimensional intensity matrix to irradiate the largest possible area in each segment. The algorithm can be applied directly to MLC systems with different motion constraints. This algorithm has been evaluated by generating 1000 random 15 x 15 cm intensity matrices, each having from 3 to 16 intensity levels. Five clinical intensity modulated fields generated from the NOMOS CORVUS planning system for a complex clinical head and neck case were also tested with this and two other algorithms. The results of both the statistical and clinical studies showed that for all the intensity matrices tested, the proposed algorithm results in the smallest number of segments with a moderately increased monitor units. Thus it is well-suited for use in static MLC intensity modulation beam delivery. For MLC systems with interleaf motion constraint, we prove mathematically that this constraint reduces the tongue and groove effect at the expense of an increase of 25% in the number of segments. PMID- 9725130 TI - Use of CEA TVS film for measuring high energy photon beam dose distributions. AB - CEA TVS film is a therapy verification film that has been recently introduced in the North American market. This film features linear characteristic curves for photon energies from 137Cs to 18 MV as reported by Cheng and Das [Med. Phys. 23, 1225 (1996)]. In Saskatoon, TVS film was investigated for its application in the measurement of dose distributions with 4 and 18 MV linacs and a 60Co unit. The TVS film jacket has a layer of conductive material that has a minimal effect on the film's response. Film sensitivity generally increases for exposures normal to the incident beam as compared with parallel exposures, but was highly dependent on beam energy and depth of measurement. Fractional depth doses obtained in the parallel orientation agreed well with ion chamber measurements for the linac beams at depths beyond Dmax; ion chamber measurements differed by a maximum of 1.6% and 2.6% for the 4 and 18 MV beams, respectively. In the buildup region, an increase in film response was found when compared to the ion chamber measurements for both linac beams. With the 60Co beam, the TVS film showed an increase in sensitivity with depth as the proportion of scattered soft x rays increases; the maximum difference between ion chamber and film fractional depth doses was 7.8%. The TVS film demonstrates a substantial improvement over Kodak X-Omat V film for measuring depth doses in the parallel orientation, for all beams considered. Generally, the results confirm TVS film as an accurate and practical dosimeter for the measurement of dose distributions in high energy photon beams. PMID- 9725131 TI - A stereotactic radiation therapy device for retinoblastoma using a noncircular collimator and intensity filter. AB - The proximity of the lens to the retina makes the treatment of retinoblastoma a challenge for external beam radiation therapy. The approximately 1 mm separation between the posterior edge of the lens and the anterior region of the retina causes a trade-off between coverage of the entire retina and excessive dose to the lens. A stereotactic, LINAC based, lens sparing technique for treating retinoblastoma is presented. The technique uses noncoplanar arcs with the lens at isocenter. A special noncircular collimator blocks the lens but it also causes the dose distribution to vary across the retina. A fluence modulation filter is used to reduce the dose inhomogeneity across the target. The resulting dose distribution is roughly hemispheric, providing both anterior coverage of the retina and lens blocking unlike conventional techniques. The method used to develop the collimator and filter assembly is presented. Dosimetry of the assembly was carried out using radiochromic film, and the results were entered in a treatment planning system. The dose distribution as measured in a phantom is provided and compared to calculations. PMID- 9725132 TI - Linear accelerator output variations and their consequences for megavoltage imaging. AB - An experimental study of radiation output intensity fluctuations of a Philips SL25 linear accelerator is presented. Measurements are obtained using an electronic portal imaging device, and the consequences of the measured fluctuations for various different applications of megavoltage imaging including portal imaging, transit dosimetry and megavoltage computed tomography (MVCT) are discussed with examples. Fluctuations in output of +/- 0.7% (1 SD) are seen on every radiation pulse after photon noise and uncertainties caused by the detection system have been accounted for. Large fluctuations are also seen during the initial beam stabilization period (15%), during normal accelerator operation after the beam has been on for more than 1 min (4.5%) and during are therapy as a repeatable function of gantry angle (9%). Such output intensity fluctuations are shown to produce image artifacts in portal imaging devices with scanned detector readout and can also produce systematic errors in detector calibration that would lead to uncertainty in transit dose calculations. The propagation of these intensity fluctuations through MVCT image reconstruction is shown to produce ring artifacts in the reconstructed image. Sample portal and MVCT images are presented. All observed fluctuations in accelerator output are well within the manufacturer's specifications and do not affect the total dose delivered during normal treatment. Finally, megavoltage imaging is shown to be a powerful tool for accelerator quality assurance and treatment verification. PMID- 9725133 TI - Measurement of Prepl Pwall factors in electron beams and in a 60Co beam for plane parallel chambers. AB - This paper describes a method to measure the product of Prepl Pwall correction factors for ionization chambers and presents our measured values of Prepl Pwall for Markus plane-parallel chambers in electron beams. It is shown that the measured values of Prepl Pwall can be fitted to an equation, Prepl Pwall = c1 + c2 R50 + c3 (R50)2, for Markus chambers at the new reference depth for electron beams (6 MeV < or = nominal energy E < or = 20 MeV). We also present our measured values of Prepl Pwall for NACP and Markus chambers in a water phantom irradiated in a 60Co beam. PMID- 9725134 TI - Tissue-air ratios for narrow 60Co gamma-ray beams. AB - This study introduces a table of tissue-air ratios (TAR) for narrow 60Co gamma ray beams. The table is consistent with recently published TAR data for broad 60Co gamma-ray beams [Table 4.1, Br. J. Radiol. Suppl. 25 (1996)]. Narrow-beam TARs are derived analytically from broad-beam data of Table 4.1 and are tabulated for circular fields ranging from 0.2 to 2.2 cm in radius--an approximate equivalent of a 0.4 cm x 0.4 cm to 4 cm x 4 cm square-field range. The extent of depth is from 0.5 to 30 cm in water. PMID- 9725135 TI - Dosimetric evaluation of a widely used kilovoltage x-ray unit for endocavitary radiotherapy. AB - In this paper we present the dosimetric data of a Therapax DTX300 kilovoltage x ray unit for endocavitary rectal irradiation. The unit if operated at tube voltage of 40-60 kVp (30 mA) with an added filtration of 0.2-0.4 mm Al generates acceptable beam qualities comparable to those of the original Papillon technique. Relative dosimetric measurements were performed at the cone end (37.2 cm SSD) of a 3 cm diameter rectal cone using various detectors to ensure the accuracy. A Monte Carlo method was used to calculate correction factors for the diode used in the percentage depth-dose (PDD) measurement, and to study the effect of the detector size on the beam profile. The PDD data were determined using the diode measurement corrected for its energy and angular response. It was found that the PTW N23342 and Markus parallel-plate chamber can be used directly to measure the PDD for this beam quality with 2% uncertainty. Measurement and Monte Carlo results have shown that the detector size has a significant effect on the penumbral profile. Film and diode detectors have a better spatial resolution compared to ionization chambers, but they may give an incorrect profile tail due to either nonlinear response at low energy or angular dependence. This can be corrected using the ionization-chamber measurement, based on the Monte Carlo analysis. The isodose distributions for this x-ray unit are presented. PMID- 9725136 TI - A statistical investigation of the scaling factor method of beta-ray dose distribution derivation: the scaling factor for water to bone. AB - Reliable methods of estimating doses are essential for the use of beta emitting radionuclides for radiotherapy. The passage of electrons through matter is a very complex phenomenon due to the large number of elastic and inelastic interactions resulting in scattering and energy losses. The analytical solution for the electron transport being intractable, the problem has been addressed by the Monte Carlo technique. Empirical or semiempirical less time consuming methods, such as the scaling factor method, may appear more preferable in practice when dealing with complicated source distributions. The method, proposed by Cross and co workers [AECL Report Nos. AECL-1617 (1982), AECL 10521 (1992)] consists in the derivation of beta-ray dose distribution in other media from those in water by using a "scaling factor" or "relative attenuation factor" on distance and a closely related renormalization factor imposed by the energy conservation. This work investigates the accuracy of the scaling factor method using a statistical approach, a generalized chi 2 test, focusing on the particular case of potential interest, the scaling factor for water to bone. The direct comparison of the shapes of the depth dose deposition curves in the two media indicates discrepancies of less than 5% up to at least 60% of the range in bone, a depth within which 95% of the initial energy is deposited. The scaling factor derived by this method, 0.9720 +/- 0.0012, confirms the existing experimentally determined value of 0.973 +/- 1% [AECL Report No. AECL-10521 (1992)]. The accuracy of the determination is increased by almost a factor of 10. A way of improving the scaling method, especially for depth over the 60% continuous slowing down approximation range, by using a modulation function is also proposed. PMID- 9725137 TI - Application of the scaling factor method to estimation of beta dose distributions for dissimilar media separated by a planar interface. AB - The most accurate method of calculating beta dose distribution currently relies on the Monte Carlo technique. The major drawback of the method is the long computing time required to follow a large number of "electron histories" in order to achieve good statistics, which makes the method unattractive for practical radiation therapy. A way to avoid the Monte Carlo calculations for homogeneous media was suggested by Cross and co-workers (AECL Report Nos. 7617, 1982; 10521, 1992), and is known as the "scaling factor" method. It consists of the determination of the depth dose distribution in a medium based on known data about the dose distribution in an arbitrary reference medium (e.g., air, water) by the use of a scaling factor on distance and a closely related renormalization factor imposed by energy conservation. This work is an attempt to extend the applicability of the scaling factor method to dissimilar media to a planar interface. The investigation was done for an isotropic source of the radioisotope 32P and an interface between water and medium "i," where medium "i" could be any medium with atomic number in the range 8 < Z < 50. The method was checked using three randomly chosen elements 40Zr, 32Ge, and 26Fe, each forming planar interfaces with water at either 100 or 350 mg/cm2. Discrepancies of less than 5% were detected (acceptable for practical radiotherapy) for the depth within which at least 95% of the initial energy is deposited. PMID- 9725138 TI - Yttrium-90 biodistribution by yttrium-87 imaging: a theoretical feasibility analysis. AB - Yttrium-90 is widely used in radioimmunotherapy but does not emit photons for imaging. Indium-111, a chemically similar atom has been used instead as a tracer for the distribution of 90Y-labeled antibodies. Recent advances in gamma camera technology have made it possible to image at energies higher than 364 keV. This work provides a theoretical analysis of the feasibility of using 87Y (485 keV, 92.2% yield) as a tracer for in vivo imaging of 90Y-labeled antibodies. Yttrium 87 may be produced by the 87Sr(p,n)87Y reaction in a cyclotron and has a 3.3 day half-life. This reaction also yields a metastable state of 87Y (87mY). Yttrium 87, itself decays to a metastable state of 87Sr (87mSr). The level of these contaminants and their anticipated impact on the utility of 87Y as a tracer for 90Y are examined theoretically. Dosimetry is performed to assess the absorbed dose associated with using 87Y. A 100 h delay following the end of bombardment of an isotopically enriched 87Sr target reduces the activity of the metastable state of 87Y in the product by 10- to 15-fold, with greater delays resulting in a further reduction. A rapid equilibration between 87Y and its daughter, 87mSr, is expected, in vivo. Although strontium is known to concentrate in bone, the electron emissions of 87mSr are short range, thereby making possible biologic effects highly position dependent; the photon emissions allow for independent imaging which may be used to perform dosimetry and thereby directly assess potential toxicity. 87Y merits further consideration as an imaging tracer for 90Y. PMID- 9725139 TI - Seed localization in eye plaque brachytherapy. AB - We describe a simple method for localizing seeds in eye plaques used for brachytherapy treatment of intraocular tumors and age-related macular degeneration. The method is based on obtaining magnified photocopy images of plaques with seeds in place and then using simple geometrical considerations to reconstruct seed coordinates with about 2% accuracy. A spreadsheet program is used for reconstruction and dosimetric calculations. PMID- 9725140 TI - Astigmatic single photon emission computed tomography imaging with a displaced center of rotation. AB - A filtered backprojection algorithm is developed for single photon emission computed tomography (SPECT) imaging with an astigmatic collimator having a displaced center of rotation. The astigmatic collimator has two perpendicular focal lines, one that is parallel to the axis of rotation of the gamma camera and one that is perpendicular to this axis. Using SPECT simulations of projection data from a hot rod phantom and point source arrays, it is found that a lack of incorporation of the mechanical shift in the reconstruction algorithm causes errors and artifacts in reconstructed SPECT images. The collimator and acquisition parameters in the astigmatic reconstruction formula, which include focal lengths, radius of rotation, and mechanical shifts, are often partly unknown and can be determined using the projections of a point source at various projection angles. The accurate determination of these parameters by a least squares fitting technique using projection data from numerically simulated SPECT acquisitions is studied. These studies show that the accuracy of parameter determination is improved as the distance between the point source and the axis of rotation of the gamma camera is increased. The focal length of the focal line perpendicular to the axis of rotation is determined more accurately than the focal length to the focal line parallel to this axis. PMID- 9725141 TI - Analysis of methods for reducing false positives in the automated detection of clustered microcalcifications in mammograms. AB - Clustered microcalcifications are often the first sign of breast cancer in a mammogram. Nevertheless, all clustered microcalcifications are not found by an individual radiologist reading a mammogram. The use of a second reader may find those clusters of microcalcifications not found by the first reader, thereby improving the sensitivity of detecting clustered microcalcifications. Our laboratory has developed a computerized scheme for the detection of clustered microcalcifications, which can act like a second reader, that is undergoing clinical evaluation. This paper concerns the feature analysis stage of the computer scheme, which is designed to remove some of the false-computer detections. We have examined three methods of feature analysis, namely, rule based (the method currently used), an artificial neural network (ANN), and a combined method. In an independent database of 50 images, at a sensitivity of 83%, the average number of false positive (FP) detections per image was: 1.9 for rule-based, 1.6 for ANN, and 0.8 for the combined method. We demonstrate that the combined method performs best because each of the two stages eliminates different types of false positives. PMID- 9725142 TI - Automated lung segmentation in digital lateral chest radiographs. AB - We are developing a fully automated computerized scheme for segmenting the lung fields in digital lateral chest radiographs. Existing computer-aided diagnostic (CAD) schemes and automated lung segmentation methods have focused exclusively on the posteroanterior view, despite the diagnostic importance of the lateral view. Information from the lateral radiograph is routinely incorporated by radiologists in their decision-making process, and thus computer analysis of lateral images may potentially add another dimension to current CAD schemes. Automated analysis of the lung fields in lateral images will necessarily require accurate segmentation. Our scheme employs an initial procedure to eliminate external and subcutaneous pixels. Global gray-level histogram analysis then allows for the identification of a range of gray-level thresholds. An iterative gray-level thresholding scheme is implemented using this range of thresholds to construct a series of binary images in which contiguous regions are identified and geometrically analyzed. Regions determined to be outside the lungs are prevented from contributing to binary images at later iterations. Adaptive local gray-level thresholding is applied along the initial contour that results from the global thresholding procedure to extend the contour closer to the true lung borders. This local thresholding method uses regions of interest of various dimensions, depending on the enclosed anatomy. Smoothing and polynomial curve fitting complete the segmentation. A database of 100 normal and 100 abnormal lateral images was analyzed. Quantitative comparison of computer-segmented lung regions with lung regions manually delineated by two radiologists indicated that 83% and 84% of normal and abnormal images, respectively, displayed segmentation contours within three standard deviations of the mean inter-radiologist contour degree-of overlap value. PMID- 9725143 TI - New classes of helical weighting algorithms with applications to fast CT reconstruction. AB - The focus of this paper is on CT helical weighting algorithms using one source rotation, or 2 pi, worth of projection data. Currently known 2 pi helical weighting algorithms include a fan-angle dependency, and do not lend themselves to fast reconstruction, for two reasons. First, it can be shown that the weight distributions present a line of discontinuity across the sinogram (projection space), which defines two separate sinogram regions. Second, the expressions for the weighting functions differ for those two regions. Accordingly, reconstruction of P different image planes (all using a given projection) requires P weightings and filterings of that projection. In this paper, it is shown that, by first generalizing the concept of the interpolation/extrapolation function used in the weighting, to the concept of distance function, and second by selecting particular classes of such distance functions, the discontinuity across the sinogram can be eliminated. By imposing specific sufficient conditions on such distance functions, single analytical expressions across the entire 2 pi sinogram are obtained. Decomposition of these particular "single" distance functions leads to two-filtering reconstruction algorithms, for which a given projection needs to be filtered only two times for an arbitrary number P of reconstruction planes. Finally, another generalization of the concept of helical weighting leads to one filtering weight functions that depend only on the sum of the projection--and fan angles. Accordingly, after rebinning the fan-beam projections to parallel projections, the corresponding 2 pi helical weighting algorithms do not include a dependency over the ray parameter. Equivalently, for these algorithms, weighting commutes with filtering, and reconstruction of an arbitrary number P of image planes requires only one filtering per projection. These algorithms are shown to be consistent with the hypothesis of a linear z variation of the projections. PMID- 9725144 TI - Lens design for large-area x-ray sensitive vidicons. AB - Large-area x-ray sensitive vidicons have the potential to be superior to conventional x-ray image intensifiers for medical fluoroscopy. To build a large area x-ray vidicon, an electron lens system is necessary to provide perpendicular beam landing. In this paper, we discuss theoretically how to build two and three cylinder coaxial lenses for this purpose. Some general design rules and a preferred design which satisfies our design criteria are presented. Limitations of these simple types of electron lens are discussed. Two advanced designs for vidicons with short lengths or reduced beam landing error are also included. Among them, the deflection-center-change system is introduced here for the first time. PMID- 9725145 TI - Comment on "Real-time B-mode ultrasound quality control test procedures" [Med. Phys. 25, 1385-1406 (1998)]. PMID- 9725146 TI - [Pain relief therapy: Oxycodone--a new retard opioid with effective analgesic action]. PMID- 9725147 TI - Pacemaker trials: software or hardware randomization? PMID- 9725148 TI - Effects of postmyocardial infarction scar size, cardiac function, and severity of coronary artery disease on QT interval dispersion as a risk factor for complex ventricular arrhythmia. AB - The aim of the study was to determine the relation between QT dispersion and ventricular arrhythmia after myocardial infarction, as well as the effects of postinfarction scar size, cardiac function, and severity of coronary artery disease on QT dispersion. Three hundred three patients, 3 months after myocardial infarction, and a group of 21 healthy subjects were evaluated. QT dispersion was the difference between maximal and minimal QT interval in 12-ECG leads. Postinfarction scar size was determined by Selvester's QRS scoring system. Cardiac function was evaluated by echocardiography and exercise stress test, and the severity of coronary artery disease by the number and degree of coronary artery stenoses. QT dispersion increased significantly in relation to the severity of arrhythmia (< 50 premature ventricular complexes vs ventricular tachycardia; 61.6 [+/- 12.3] vs 84.8 [+/- 16.4] ms, P < 0.001). QT dispersion > 80 ms was associated with ventricular tachycardia with the sensitivity of 68% and specificity of 88%. QT dispersion also increased significantly, dependent on the postinfarction scar size (0% vs > or = 33% of left ventricular myocardium; 61.8 [+/- 16.4] vs 74.7 [+/- 16] ms, P < 0.001), as well as in the case of significantly impaired cardiac function. Although QT dispersion increased with the number of diseased vessels and the degree of stenoses, the differences were not significant (P > 0.05). In conclusion, QT dispersion is a risk marker of complex ventricular arrhythmia in the chronic stage of myocardial infarction. Multiple regression analysis indicates that only the postinfarction scar size has an independent effect on QT dispersion (R2 = 0.39, P < 0.05). PMID- 9725150 TI - Atrial lead implantation during atrial flutter or fibrillation? AB - In patients with sinoatrial disease, unexpected atrial flutter (Af) or fibrillation (AF) is a common problem during implantation of atrial-based pacing systems. As an alternative approach to blind atrial lead placement, lead positioning could be optimized by atrial electrogram mapping. It was the object of this study to evaluate if atrial lead implantation according to this approach and during ongoing arrhythmia is reasonable or if it should be postponed until restoration of sinus rhythm (SR). Twenty-nine consecutive patients (group I) with sick sinus syndrome received a dual-chamber pacemaker during an episode of Af (n = 11) or AF (n = 18). All but two atrial leads were of the screw-in type and had bipolar sensing. Atrial lead position was optimized by mapping the electrogram under fluoroscopy to find locations with high potential amplitudes. The patients were followed for 15.1 +/- 9.8 months, and atrial sensing threshold (AST), atrial pulse width threshold (PWT) at 2.0 V, the pacing mode programmed, and the clinical outcome (OUT) were recorded. The control group consisted of 30 patients (group II) who equally had a history of AF or Af, but were in SR during implantation. The atrial peak-to-peak potential (APEAK) after final lead placement was lower for AF (median value 2.5 mV, lower-upper quartile: 1.7-3.1 mV) as compared to Af (3.8 mV, 2.7-4.9 mV, P < 0.05) and SR (4.1 mV, 3.3-6.2 mV, P < 0.001). There was a correlation (P < 0.01) between APEAK during Af/AF and the postoperative AST immediately after restoration of SR. No lead in any group had to be corrected due to improper sensing in the postoperative course. Median chronic AST was 2.8 mV (2.0-4.0 mV) in group I and 4.0 mV (2.8-4.0 mV) in group II. Median chronic PWT at 2.0 V was 0.15 ms (0.12-0.26 ms) in group I and 0.15 ms (0.09-0.20 ms) in group II. There was no significant difference in chronic AST and PWT between both groups. All but two patients in group I preserved SR as the basic rhythm. A stable SR was observed in 10 of 29 patients, intermittent Af/AF was documented in 17 of 29 patients, seven of whom were asymptomatic. There was no significant difference in OUT between group I and II. Hence, sinus rhythm is not a prerequisite of atrial lead implantation. Mapping the Af or AF waves appears to be useful to guide lead placement and to achieve sufficient sensing and pacing conditions after conversion to sinus rhythm. PMID- 9725149 TI - Stent dilation of superior vena cava and innominate vein obstructions permits transvenous pacing lead implantation. AB - The purpose of this study was to assess the feasibility of stent dilation of venous obstructions/occlusions to permit transvenous pacing lead implantation. Innominate vein or superior vena cava (SVG) obstruction may preclude the implantation of transvenous pacing leads. Patients with d-transposition of the great arteries, after a Mustard or Senning procedure, and children with previously placed transvenous pacing leads are at higher risk for this vascular complication. From May 1993 to January 1996, eight pediatric patients who underwent transvenous pacing lead implantation or replacement were found to have significant innominate vein or SVC obstruction or occlusion. Utilizing intravascular stents, a combined interventional and electrophysiological approach was used to relieve the venous obstruction and to permit implantation of a new transvenous pacing lead. Two patients had complete SVC occlusion requiring puncture through the obstruction with a transseptal needle. Vessel recanalization was achieved with balloon dilation and stent implantation. The remaining six patients had severe venous obstruction with a mean minimum diameter of 3.1 +/- 3.3 mm. The mean pressure gradient across the obstructed veins was 8.6 +/- 7.3 mmHg. Following implantation of 15 Palmaz P308 stents in eight vessels, the mean diameter increased to 14.2 +/- 1.9 mm and the mean pressure gradient across the stented vessels decreased to 1.0 +/- 2.0 mmHg. A transvenous pacing lead was implanted successfully through the stent(s) immediately or 6-8 weeks later. Innominate vein and SVC obstruction can be safely and effectively relieved with intravascular stents and permit immediate or subsequent transvenous pacing lead implantation. PMID- 9725151 TI - Usefulness of plasma catecholamines during head-up tilt as a measure of sympathetic activation in vasovagal patients. AB - Vasovagal syncope is a common clinical disorder which has been traditionally related to a vasovagal reflex precipitated by an initial excess sympathetic stimulation. We hypothesized that the increase in plasma catecholamines during head-up tilt is more accentuated in patients with tilt induced vasovagal syncope. To test this hypothesis, plasma catecholamines were measured in supine posture and during head-up tilt in patients with a history suggestive of vasovagal syncope. Of these, 13 had a normal response to tilt (nonvasovagal group; age 41 +/- 19 [SD]years) and 11 had a vasovagal response to tilt (vasovagal group; 39 +/ 20 years). In the supine posture at rest, plasma epinephrine and norepinephrine were not significantly different between the nonvasovagal and the vasovagal groups (39 +/- 28 ng/L vs 46 +/- 38 ng/L, P = 0.5792, 335 +/- 158 ng/L vs 304 +/- 124 ng/L, P = 0.6007, respectively). Furthermore, the tilt induced changes in plasma epinephrine and norepinephrine were not different between the two groups (20 +/- 20 ng/L vs 35 +/- 55 ng/L, P = 0.3562, 264 +/- 158 ng/L vs 242 +/- 205 ng/L, P = 0.7724, respectively) suggesting that differences in the hemodynamic response to tilt are not predictable by the supine levels of circulating plasma catecholamines, and that the extent of plasma catecholamines increase during tilt does not determine the hemodynamic outcome of the tilt test. Since orthostatic changes of plasma catecholamines could be influenced by volume factors, we assessed plasma renin activity and aldosterone as surrogates of blood volume. Baseline plasma renin activity and aldosterone were not significantly different between the two groups. We conclude that inasmuch as plasma catecholamines reflect the status of sympathetic activity, our data do not support the hypothesis that accentuation of sympathetic activity precedes necessarily the tilt induced vasovagal syncope. However, one should take in consideration that multiple factors may influence catecholamine levels and catecholamines kinetics. A hyperresponsiveness of beta-receptors to catecholamines in patients with vasovagal syncope may be suggested but needs to be tested. PMID- 9725153 TI - Koch's triangle sized up: anatomical landmarks in perspective of catheter ablation procedures. AB - Koch's triangle contains the compact part of the atrioventricular node and is anatomically delineated by the Eustachian ridge, the membranous septum, and the insertion of the tricuspid valve. Inferiorly, Koch's triangle ends at the site of the os of the coronary sinus and, in part, is continuous with the sub-Eustachian pouch. Catheter ablation procedures for several types of reentrant tachycardias are based on identifying these anatomical landmarks. Variability in the dimensions of Koch's triangle thus may be clinically relevant. We examined 50 hearts. Anatomical landmarks measured were the Eustachian ridge, the tricuspid valve, the overall length between the membranous septum and the coronary sinus, the width of the coronary sinus, the Eustachian ridge in its roof and the distance to the tricuspid valve, and that of the sub-Eustachian pouch. Individual variations were marked. The mean values (+/- SD) were: Eustachian ridge 29.4 +/- 5.3 mm, tricuspid valve 28.9 +/- 4.5 mm, coronary sinus 10.8 +/- 2.2 mm, Eustachian ridge 3.7 +/- 2.3 mm, space underneath coronary sinus 8.6 +/- 3.4 mm, and sub-Eustachian pouch 26.8 +/- 3.3 mm. The overall length varied between 15 and 38 mm, with a mean of 26.3 +/- 4.5. In conclusion, Koch's triangle shows considerable individual variations in size. Given the fact that the absolute figures for the range in size of the compact atrioventricular node is much less than that of Koch's triangle, these variations have implications for catheter ablation procedures. PMID- 9725152 TI - Association between cardiac autonomic function and coping style in healthy subjects. AB - The link between personality and cardiac function is insufficiently characterized. We postulated that in a healthy population, cardiac autonomic function is linked to coping style. In 276 healthy volunteers, between the ages of 18 and 71, the Utrecht Coping List was used to evaluate different coping strategies. Trait anxiety was scored by the Spielberger State Trait Anxiety Inventory. A 24-hour Holter recording was used to calculate heart rate variability (HRV). For HRV parameters and coping mechanisms this study demonstrated gender-specific differences and correlations with age. In men (n = 141) higher active coping was associated with less global autonomic activity or SDANN (rs = -0.27, P < 0.001). This relationship was most prevalent in young (18 30 years) men (rs = -0.45, P < 0.005). Higher expression of negative emotions or anger was related to both higher vagal (rs = 0.23 for rMSSD, P < 0.01) tone and higher LF power (rs = 0.23, P < 0.01). In young men expression of negative emotions or anger was associated with LF power (rs = 0.37, P < 0.01) and in middle-aged (31-50 years) men with vagal tone (rs = 0.43 for rMSSD, P < 0.005) and heart rate (rs = -0.41, P < 0.005). Higher comforting ideas was related to higher LF power (rs = 0.23 for LF power, P < 0.005), and this especially in middle-aged men (rs = 0.37, P < 0.01). In women (n = 135), no significant correlations between coping style and HRV indices were found. We conclude that in normal individuals, at least in men, our findings suggest a relationship between coping style and cardiac autonomic function. PMID- 9725154 TI - An integrated dual sensor system automatically optimized by target rate histogram. AB - The use of combined sensors and advanced algorithms using different principles can improve rate performance over a single sensor system. Combinations of sensors and more sophisticated algorithms, however, invariably increase the complexity of pacemaker programming. An automatically optimized combined minute ventilation and activity DDDR pacemaker was developed to minimize repeated sensor adjustment. The device used subthreshold (below cardiac stimulation threshold) lead impedance to detect lead configuration at implantation automatically, followed by "implant management," including setting of lead polarity and initiation of DDDR pacing. Automatic sensor adaptation was achieved by programming a "target rate histogram" based on the patient's activity level and frequency of exertion, and the rate profile optimization process matched the recorded integrated sensor response to the target rate histogram profile. In nine patients implanted with the DX2 pacemakers, the implant management gave 100% accuracy in the detection of lead polarity. Rate profile optimization automatically increased the pacing rate during exercise between discharge and 3-month follow-up (hall walk: 78 +/- 3 vs 98 +/- 3 beats/min, and maximal treadmill exercise: 89 +/- 6 vs 115 +/- 5 beats/min, P < 0.001) with a significant increase in exercise duration during maximal exercise (7.18 +/- 1 min vs 9.56 +/- 2 min, P = 0.05). The accuracy of rate profile optimization versus manual programming was assessed at 1 month, and there was no significant difference between pacing rate kinetics and maximal pacing rate between the two methods of programming. In conclusion, pacemaker automaticity can be initiated at implantation and the self-optimized rate adaptive response appeared to be comparable to that derived from a manual programming procedure, which may reduce the need to perform time consuming sensor programming. PMID- 9725155 TI - Automatic adjustment of pacemaker stimulation output correlated with continuously monitored capture thresholds: a multicenter study. European Microny Study Group. AB - Pacing threshold is affected by many factors. A pacing system able to confirm capture at each beat and automatically adjust its output close to the actual pacing threshold is highly desirable. This study evaluates the safety and efficacy of the Autocapture function of the Pacesetter Microny SR+. One hundred thirteen patients were recruited from 16 centers in 7 European countries and followed up for 1 year. All pacemakers were implanted with Pacesetter's low polarization, bipolar leads. The key feature of Autocapture is the immediate delivery of a 4.5 V safety backup pulse 62.5 ms after any ineffective ongoing low output pulse. Holter recordings confirmed total reliability of this feature without any exit block. The measured evoked response (ER) signal was stable over time. Acute and chronic pacing thresholds measured by VARIO and Autocapture tests correlated (r > 0.79) over the period of the study. The incidence of backup pulses was 1.1% during pacing. With Autocapture programmed ON, the overall total current consumption was 4.1 microA for VVI and 5.0 microA for VVIR pacing. This study proved that the Autocapture safely and reliably regulates the pacemaker's output according to the prevailing threshold thus providing maximum patient safety and prolonging service life. PMID- 9725156 TI - Koch's triangle in pediatric age: correlation with extra- and intracardiac parameters. AB - The atrioventricular node is situated in the lower atrial septum, at the apex of the Koch's triangle. The dimensions of the Koch's triangle are studied in adult humans, while no data exist about them in pediatric age. The knowledge of the dimensions of Koch's triangle in childhood is very important for safe and correct application of radiofrequency energy during transcatheter ablation. The dimensions of Koch's triangle were determined in 69 human pediatric hearts. The median age of the children was 3 months, with a range from 1 day to 14 years, 30 were female and 39 were male. Relations between body weight (extracardiac parameter) and tricuspid valve diameter (intracardiac parameter) were determined in all hearts to show morphometric modifications with growth. The distribution of body weight was not Gaussian and no correlation could be obtained between Koch's triangle dimensions and body weight. However, it was possible to identify that the mean ratio between the cathetus of the Koch's triangle corresponding to the annulus of the tricuspid valve and the tricuspid valve diameter was 0.45 +/- 0.16, with a highly significant correlation coefficient (r = 0.653, P < 0.001). Therefore, by knowing: (1) the diameter of the tricuspid valve, and (2) the constant ratio between the cathetus of the Koch's triangle and the tricuspid valve diameter, it is possible to calculate the length of the segment of the tricuspid annulus along which the transcatheter application of radiofrequency current can be applied to ablate the slow-pathway, thus reducing the risks of damage of the atrioventricular node. PMID- 9725157 TI - Sudden Wenckebach periods and their relationship to neurocardiogenic syncope. AB - Throughout a 9-month period during which 1,125 Holter tapes were reviewed prospectively we identified 13 nonmedicated patients with an arrhythmia, which for the purposes of this presentation was categorized, because of their mode of initiation, as sudden Wenckebach periods (WP). The episodes emerged abruptly from a normal (< or = 200 ms) PR interval with sudden prolongation of PR and PP intervals (and reversed PR-RP relationship) that took place over 1-8 cycles. The postpaced PR interval was shorter than that of the last conducted beat. The episodes were separated into two groups. Group I included 11 patients with symptoms other than syncope and Group II included 2 patients with syncope. There were 26 episodes of sudden WP in Group I. Twenty-five terminated in a single (and one in double) blocked P waves. Most episodes occurred between 10 PM and 7 AM. Symptoms did not correlate with the episodes. Mean 24-hour rates were < 90. In Group II there were 22 episodes, all occurring between 6 AM and 10 PM. The mean sinus cycle lengths before the phenomenon started to occur in Group I (861 +/- 185 ms) as well as the cycle lengths at the onset of block (1,096 +/- 215 ms) were statistically longer than those in Group II (591 +/- 40 ms and 747 +/- 63 ms, respectively, P < 0.0001). Although the mode of onset in the episodes in Group II was similar to Group I, 16 episodes terminated in 2-6 blocked P waves. Thus, the entire number of episodes could be categorized as an unusual type (because of the PR prolongation) of paroxysmal, or advanced second degree AV block. Because these patients had negative electrophysiological studies, positive tilt tests, and absent syncope after oral propranolol therapy, they were considered as having neurocardiogenic syncope. In addition, the faster than normal (> 100) mean 24-hour rates) suggested that they also had so-called inappropriate sinus tachycardia. In summary, Group I consisted of patients with a normal, benign, vagal-induced second-degree AV block, whereas the Holter findings in Group II appeared to reflect unusual (but natural, i.e., nonprovoked) electrocardiographic manifestations of certain patients with neurocardiogenic syncope. PMID- 9725158 TI - QRS duration widening: reduced synchronization of endocardial activation or transseptal conduction time? AB - Antegrade activation of the His-Purkinje system (HPS) results in synchronized activation of the right ventricular (RV) and left ventricular (LV) endocardia forming normal, narrow QRS duration (QRSD). An alteration in septal activation and transseptal conduction time have been reported to be the causes for QRSD widening seen with bundle branch block. However, reduced synchronization of activation of RV and LV endocardia as another potential mechanism for QRSD widening has not been systematically studied. Fifteen consecutive patients underwent radiofrequency ablation (RFA) for treatment of supraventricular tachycardia. After RFA, mean QRSD in normal sinus rhythm was 86 +/- 8 ms with mean HV interval of 40 +/- 5 ms. Right atrial (RA), coronary sinus (CS), simultaneous (S) RA-CS, RV apex (RVA), LV apex (LVA), and SRVA-LVA pacing were performed. Mean QRSD with RA, CS, SRA-CS pacing was similar to normal sinus rhythm (87 +/- 7, 87 +/- 8 and 88 +/- 8 ms respectively). Mean QRSD was significantly longer with SRVA-LVA and either RVA or LVA pacing alone compared to normal sinus rhythm (106 +/- 8, 146 +/- 12 and 157 +/- 13 ms, respectively). However, QRSD was significantly shorter with SRVA-LVA pacing compared to either RVA or LVA pacing alone (P < 0.0001). We conclude that shorter QRSD with SRVA-LVA pacing compared to either RVA or LVA pacing alone is due to elimination of transseptal conduction delay; longer QRSD with SRVA-LVA pacing compared to sinus or atrial paced rhythm is due to reduced synchronization of endocardial activation secondary to ectopic entry of impulses into the HPS network and inability to take advantage of the branching structure of the HPS. Therefore, in addition to transseptal conduction delay, reduced synchronization of endocardial activation is another potential mechanism for QRSD widening. PMID- 9725159 TI - Effects of left atrial dilatation on the endocardial atrial defibrillation threshold: a study in an ovine model of pacing induced dilated cardiomyopathy. AB - Left atrial (LA) dilation is a common finding in patients with chronic atrial fibrillation (AF). Progressive dilatation may alter the atrial defibrillation threshold (ADFT). In our study, epicardial electrodes were implanted on the LA free wall and right ventricular apex of eight adult sheep. Large surface area, coiled endocardial electrodes were positioned in the coronary sinus and right atrium (RA). LA dilatation was induced by rapid ventricular pacing (190 beats/min) for 6 weeks and echocardiographically assessed weekly along with the ADFT (under propofol anesthesia). LA effective refractory period (ERP) was measured every 2-3 days using a standard extra stimulus technique and 400 ms drive. The AF cycle length (AFCL) was assessed from LA electrograms. During the 6 weeks of pacing the mean LA area increased from 6.1 +/- 1.5 to 21.3 +/- 2.4 cm2. There were no significant changes in the mean ADFT (122 +/- 15 V), circuit impedance (46 +/- 5 omega), or LA AFCL (136 +/- 23 ms). There was a significant increase in the mean LA ERP (106 +/- 10 ms at day 0, and 120 +/- 13 ms at day 42 of pacing). In this study, using chronically implanted defibrillation leads, the minimal energy requirements for successful AF were not significantly altered by ongoing left atrial dilatation. This finding is a further endorsement of the efficiency of the coronary sinus/RA shock vector. Furthermore, the apparent stability of the AF present may be a further indication of a link between the type of AF and the ADFT. PMID- 9725160 TI - Should unipolar leads be implanted in the atrium? A Holter electrocardiographic comparison of threshold adapted unipolar and high sensitive bipolar sensing. AB - The accuracy of atrial sensing plays a central role in dual chamber pacing. Recent Holter electrocardiographic studies showed a high incidence of atrial malsensing. We investigated the efficacy of bipolar atrial sensing at high sensitivity compared to threshold adapted unipolar sensing. One hundred consecutive patients with identical dual chamber pacemakers and bipolar atrial leads were investigated. Mean and individual range of 40 unipolar and bipolar telemetered atrial potentials were calculated; sensing threshold was determined by a semiautomatic sensing test. Oversensing was investigated with the help of a muscle provocation test. Twenty-four-hour Holter monitoring was performed at the highest bipolar sensitivity as well as at a unipolar sensitivity of half the measured sensing threshold. Mean atrial potential was significantly lower during bipolar mode compared to the unipolar sensing configuration, 3.66 +/- 1.75 versus 3.85 +/- 1.62 mV, P = 0.02. The bipolar atrial potentials showed a higher individual range than the unipolar signals, 2.44 +/- 2.62 versus 1.79 +/- 0.92 mV, P < 0.01. Sensing threshold did not differ significantly, 2.76 +/- 1.33 versus 2.67 +/- 1.29 mV. Mean oversensing threshold was 1.21 mV at unipolar configuration, whereas oversensing could not be provoked at a bipolar sensitivity of 0.5 mV. The incidence of atrial undersensing was significantly higher at threshold adapted unipolar sensing compared to bipolar sensing at highest atrial sensitivity, 35% versus 22%, P = 0.04. Oversensing did not occur at bipolar sensing, but was observed in 56% of patients at unipolar mode. Thirty-two percent of patients showed both atrial undersensing and oversensing at the unipolar sensing configuration. The muscle provocation test reached a sensitivity of 89% and a specificity of 95% in prediction of atrial oversensing during daily life. In conclusion, unipolar atrial potentials are more stable than bipolar ones. On the other hand, bipolar atrial sensing is less prone to the perception of myopotentials. Programming a high bipolar sensitivity significantly improves atrial sensing. Thus, bipolar leads should generally be implanted in the atrium. PMID- 9725161 TI - Intracardiac QRS electrogram width--an arrhythmia detection feature for implantable cardioverter defibrillators: exercise induced variation as a base for device programming. AB - Delivery of inappropriate therapy of implantable cardioverter defibrillators (ICD) due to inaccurate arrhythmia detection represents a major clinical problem. Different arrhythmia detection criteria such as the "stability" of the cycle length or the suddenness of "onset" of tachycardia have been implemented in ICD software to prevent inappropriate therapy. The new Medtronic model 7223Cx ICD offers an additional detection parameter (QRS width), which reflects changes in the duration of ventricular depolarization as a tool to distinguish supraventricular from ventricular tachycardias. Although this criterion can be programmed based on ECG parameters derived from resting ECGs, this may not be sufficient since QRS width is subject to considerable changes due to transient myocardial ischemia, changes in autonomic tone, or frequency dependent effects of antiarrhythmic drugs. The present study aimed to determine frequency dependent changes in QRS width in individual patients at rest and during symptom-limited exercise testing in 16 patients with documented ventricular tachycardia (N = 13) or ventricular fibrillation (N = 3). The optimal EGM slew threshold and the individual variation of QRS width were determined. Measurements obtained at the end of the implantation procedure were compared to those performed at hospital discharge. The majority of patients showed a wider variation in QRS duration as measured from 30 consecutive cycles during exercise as compared to rest. For example, the QRS range (i.e., the difference between the maximal and the minimal QRS width measured) averaged 7 +/- 3 ms at rest and increased to 11 +/- 3 ms during exercise (P = 0.004) with an increase of > or = 4 ms observed in 11 (69%) of 16 patients. In 13 (81%) of 16 patients a reprogramming of at least one QRS width parameter from its value at the time of implantation was necessary. Thus, the QRS width measured from the intracardiac EGM shows significant intraindividual variations in different physiological conditions. For optimal programming of the QRS width parameter, measurements obtained during exercise are important. PMID- 9725162 TI - Reversion to back-up mode (VOO) in a DDD pacemaker model. AB - Sorin recently identified mode reversion to backup mode (VOO). They estimate a prevalence of 1% and suggest electromagnetic interference (EMI) as the cause. We identified ten patients being followed in our pacing clinic, of whom four were found to have suffered mode reversion. We, therefore, conclude that the prevalence of this problem may be greater than predicted. None of the patients who were found to have mode reversion gave any history to suggest that they had been exposed to EMI. PMID- 9725163 TI - Magnetic electroanatomical mapping for ablation of focal atrial tachycardias. AB - Uniform success for ablation of focal atrial tachycardias has been difficult to achieve using standard catheter mapping and ablation techniques. In addition, our understanding of the complex relationship between atrial anatomy, electrophysiology, and surface ECG P wave morphology remains primitive. The magnetic electroanatomical mapping and display system (CARTO) offers an on-line display of electrical activation and/or signal amplitude related to the anatomical location of the recorded sites in the mapped chamber. A window of electrical interest is established based on signals timed from an electrical reference that usually represents a fixed electrogram recording from the coronary sinus or the atrial appendage. This window of electrical interest is established to include atrial activation prior to the onset of the P wave activity associated with the site of origin of a focal atrial tachycardia. Anatomical and electrical landmarks are defined with limited fluoroscopic imaging support and more detailed global chamber and more focal atrial mapping can be performed with minimal fluoroscopic guidance. A three-dimensional color map representing atrial activation or voltage amplitude at the magnetically defined anatomical sites is displayed with on-line data acquisition. This display can be manipulated to facilitate viewing from any angle. Altering the zoom control, triangle fill threshold, clipping plane, or color range can all enhance the display of a more focal area of interest. We documented the feasibility of using this single mapping catheter technique for localizing and ablating focal atrial tachycardias. In a consecutive series of 8 patients with 9 focal atrial tachycardias, the use of the single catheter CARTO mapping system was associated with ablation success in all but one patient who had a left atrial tachycardia localized to the medial aspect of the orifice of the left atrial appendage. Only low power energy delivery was used in this patient because of the unavailability of temperature monitoring in the early version of the Navistar catheter, the location of the arrhythmia, and the history of arrhythmia control with flecainide. No attempt was made to limit fluoroscopy time in our study population. Nevertheless, despite data acquisition from 120-320 anatomically distinct sites during global and more detailed focal atrial mapping, total fluoroscopy exposure was typically < 30 minutes and was as little as 12 minutes. The detailed display capabilities of the CARTO system appear to offer the potential of enhancing our understanding of atrial anatomy, atrial activation, and their relationship to surface ECG P wave morphology during focal atrial tachycardias. PMID- 9725164 TI - Use of a real-time three-dimensional magnetic navigation system for radiofrequency ablation of accessory pathways. AB - Using conventional technology, accessory pathway ablation often requires prolonged exposure of the team and patient to ionizing radiation. Further, although the primary success rate (approximately 90%) and the rate of recurrence (approximately 10%) are acceptable, there is room for improvement. Finally, inadvertent ablation of the compact node and AV/His-Purkinje system still occurs particularly with septal accessory pathways. The Biosense CARTO Nonfluoroscopic Mapping and Navigation System (CARTO System) when used to locate the accessory pathway and guide delivery of radio frequency energy to the accessory pathway, has the potential to reduce radiation exposure, improve primary ablation success, and reduce the rate of recurrence and improve safety. This article describes our experience with the CARTO Biosense System relating to setting up the CARTO System specifically for WPW mapping/ablation, and features of the CARTO System, which are particularly advantageous for mapping and ablation of accessory pathways. PMID- 9725165 TI - NASPE expert consensus statement: physician workforce in cardiac electrophysiology and pacing. NASPE task force, Washington, D.C. PMID- 9725166 TI - Human precision of operating a digitizing board: implications for electrocardiogram measurements. AB - To investigate the precision achieved by human measurement on a digitizing board, 100 healthy volunteers (46 women, mean age 36 +/- 12 years) were asked to measure 15 times on artificial pattern composed of 15 points. A high precision digitizing board (programmed to the technical accuracy of +/- 50 microns) was used, and mean and maximum errors in measuring the same distance repeatedly and relocalizing the same point repeatedly were obtained for each volunteer. A median mean and maximum error of 0.2 mm and 1.0 mm were found for repeated distance measurement. When simulating QT dispersion measurement (measuring the same distance 12 times), median value of 20 ms was obtained for ECGs of 25 mm/s paper speed. The study concludes that human precision of operating a digitizing board is rather poor. A recommendation is given to use either a computer screen for manual measurement of ECGs or to provide an operator of the digitizing board with an immediate feedback of the precision and measurement stability achieved so that erroneous measurement can be actively rejected. PMID- 9725167 TI - Amiodarone induced pulmonary fibrosis in infancy. AB - We report a case of pulmonary fibrosis in an infant receiving amiodarone for treatment of intractable atrioventricular reentrant tachycardia secondary to Wolff-Parkinson-White syndrome. At 9 months, a screening chest radiograph showed a diffuse interstitial infiltrate in an asymptomatic, thriving infant. Amiodarone was discontinued and the pulmonary fibrosis resolved gradually over 6 months. This case documents the first report of amiodarone induced pulmonary fibrosis in the pediatric age group. We speculate that as amiodarone is used more frequently to manage pediatric arrhythmias, pulmonary fibrosis, a known complication of this antiarrhythmia in adults may be seen with increasing frequency in children. PMID- 9725168 TI - Radiofrequency ablation of idiopathic left ventricular tachycardia with changing ECG morphology. AB - Idiopathic left ventricular tachycardia is a distinct clinical entity with a typical ECG of right bundle branch block and left axis deviation. We presented a 39-year-old man with idiopathic left ventricular tachycardia, which demonstrated change in the configuration of QRS complex during successive radiofrequency catheter ablation. We proposed that this idiopathic left ventricular tachycardia may have alternative pathways within the reentrant circuit leading to different exits. PMID- 9725169 TI - Triple chamber pacemaker for end-stage heart failure in a patient with a previously implanted automatic defibrillator. AB - A 63-year-old man with an ischemic dilated cardiomyopathy previously implanted with an implantable cardioverter defibrillator (ICD) received a triple chamber pacemaker as an ultimate therapeutic resort for end-stage congestive heart failure. After implant, the tolerance to physical exercise increased and NYHA class decreased from III to II. Echocardiography assessed ventricular contraction resynchronization during DDD biventricular pacing as compared to VVI pacing. No major pacemaker-ICD interaction was noted during testing or follow-up. We conclude that sequential biventricular pacing is feasible in the presence of an ICD. PMID- 9725170 TI - Asymptomatic superior vena caval occlusion: a complication of nonthoracotomy implantation of cardioverter defibrillator. AB - A patient with ischemic dilated cardiomyopathy and history of ventricular fibrillation received an implantable cardioverter defibrillator by the nonthoracotomy approach. Four years later, during elective replacement of an exhausted pulse generator, a superior vena caval thrombotic occlusion with collateral circulation through the azygos and emiazygos vein systems was documented. This occlusion occurred despite an anticoagulant treatment in the standard therapeutic range. We speculate that thrombotic occlusion might be secondary to a mechanical vessel injury. PMID- 9725171 TI - The field of research in cardiology. PMID- 9725172 TI - Underdetection of ventricular tachycardias. PMID- 9725173 TI - Characterisation of recently emerged multiple antibiotic-resistant Salmonella enterica serovar typhimurium DT104 and other multiresistant phage types from Danish pig herds. AB - A total of 670 isolates of Salmonella enterica were isolated from Danish pig herds, phage typed and tested for susceptibility to amoxycillin + clavulanate, ampicillin, colistin, enrofloxacin, gentamicin, neomycin, spectinomycin, streptomycin, tetracyclines, and trimethoprim + sulphadiazine. S enterica serovar typhimurium (S typhimurium) isolates resistant to ampicillin, streptomycin and tetracycline and three isolates of S typhimurium DT104, two from 1994 and one from 1995, were further tested for resistance against chloramphenicol and sulphonamide and analysed by pulsed-field gel electrophoresis (PFGE) using the restriction enzyme Xba I. Overall, 66 per cent of the 670 isolates were sensitive to all the antimicrobial agents tested. Eleven isolates of S typhimurium were resistant to ampicillin, streptomycin and tetracycline and also resistant to other antibiotics in different resistance patterns. Seven different multiresistant clones were identified. The most common clones were four isolates of DT104 and three isolates of DT193. Two of the three S typhimurium DT104 from 1994 and 1995 were sensitive to all the antimicrobials tested whereas the remaining isolate from 1994 was resistant to spectinomycin, streptomycin and sulphonamides. All three isolates showed PFGF profiles identical to the four multiresistant DT104 isolates. Compared with most other countries antimicrobial resistance among S enterica isolated from Danish pig herds is uncommon. However, several different multiresistant clones were found. PMID- 9725175 TI - Diagnostic reliability of clinical signs in cows with suspected bovine spongiform encephalopathy. AB - The clinical findings in 50 cows with suspected and subsequently confirmed bovine spongiform encephalopathy (BSE) (group A) were compared with the clinical signs in 22 cows with suspected BSE, but with no histological evidence of the disease (group B). The chi-square test for association was used to compare the frequencies with which diagnostic signs or combinations of signs, were positive in the cows of groups A and B. When the frequency of a sign differed significantly, its sensitivity, specificity, efficiency and positive and negative predictive values were calculated. With respect to changes in behaviour the cows in group A more frequently showed increased excitability, nervous ear and eye movements, increased salivation and increased licking of the muzzle than the cows of group B. With respect to changes in sensitivity the cows in group A were more frequently hypersensitive to touch, noise and light than the cows of group B. With respect to changes in locomotion the cows in group A were more frequently ataxic than the cows in group B. PMID- 9725174 TI - A comparison of DNA amplification, isolation and serology for the detection of Chlamydia psittaci infection in cats. AB - Chlamydia psittaci is a significant cause of conjunctivitis in cats, but can be difficult to diagnose owing to the small number of organisms in conjunctival swabs. In the United Kingdom laboratory diagnosis is based on three techniques: isolation of the infectious organism, amplification of chlamydial DNA by the polymerase chain reaction (PCR) or the detection of anti-chlamydial antibodies by immunofluorescence assay. To determine the most sensitive method these techniques were compared in the field. The PCR based on previously published protocols was less sensitive than isolation, but by modifying the protocol its sensitivity was increased by a factor of 25 to 1250 and it was then more sensitive than isolation. The modified PCR detected chlamydia in samples containing non infectious organisms. Serology was of limited use in predicting which cats shed C psittaci although seronegative cats were negative by PCR and isolation. The modified PCR was the most sensitive and robust method for confirming C psittaci infection in cases of conjunctivitis in pet cats. PMID- 9725177 TI - Feline skin granuloma associated with Mycobacterium avium. PMID- 9725176 TI - Antibodies to orthopoxvirus in domestic cats in Norway. AB - The prevalence of antibodies to orthopoxvirus in 217 sera collected from domestic cats in the western part of Norway was 10.1 per cent as measured by a competitive ELISA. In one of the seropositive cats antibodies were also detected by an immunofluorescence assay. The average age of the cats sampled was 4.9 years, but the average age of the seropositive individuals was 7.3 years, higher than the average age of clinical cowpox virus cases in Britain (4.2 years), and in Germany (3.9 years). Antibodies against feline immunodeficiency virus (FIV) were detected in nine of 30 (30 per cent) of the seropositive cats, and in five of 30 (17 per cent) of the seronegative cats, which suggests that FIV infection may influence the susceptibility of domestic cats to orthopoxvirus, or vice versa. Orthopoxvirus infections, have recently been detected in rodent populations in several areas of Norway, and the infection may therefore be present in cats all over the country; cat owners and animal handlers should be aware of this (re)emerging zoonosis. PMID- 9725178 TI - Antimicrobial activity in vitro and in vivo of a canine ear cleanser. PMID- 9725179 TI - Foreign body intestinal perforation and intra-abdominal abscess formation as a complication of enteroplication in a dog. PMID- 9725180 TI - Reasons for the euthanasia of dogs and cats. PMID- 9725181 TI - Quality control in laboratory testing. PMID- 9725182 TI - Neuronal vacuolation in young rottweilers. PMID- 9725183 TI - Prognostic indicators for toxic mastitis in dairy cows. AB - During a three-year study, 54 cows with toxic mastitis were examined and a number of clinical and laboratory measurements were taken. Twenty-five (46.3 per cent) of the cows died, and in comparison with those which survived, they had a significantly higher packed cell volume (PCV) (P < 0.01), longer eyelid skin tent time (P < 0.01) and lower rectal temperature (P < 0.01). In a model designed to predict the probability of survival, these variables correctly predicted survival in 84 per cent of cases and death in 73 per cent of cases. The cows with toxic mastitis had a significantly higher PCV than a normal cohort of cows sampled at the end of the study. PMID- 9725184 TI - Study of leg weakness in two commercial broiler flocks. AB - The major causes of leg weakness/lameness were investigated in two male commercial broiler flocks. The numbers of dead and lame birds culled from the flocks each day were recorded by the flock managers. Forty-four lame birds and 22 sound birds were examined postmortem during a period of six weeks and the proximal and distal end of each femur, tibiotarsus and tarsometatarsus were examined histologically. Attempts were made to isolate bacteria and viruses from the proximal end of each femur. Blood samples were examined for antibodies to chicken anaemia virus (CAV), infectious bursal disease virus (IBDV) and Mycoplasma species. Bacterial chondronecrosis with osteomyelitis was identified in the proximal end of the femur of eight of the 44 lame birds, and in the proximal end of the tibiotarsus of a further bird (20.4 per cent). Gram-positive bacteria were present in all the lesions. Staphylococcus aureus was recovered from 62.5 per cent of the lesions confirmed by histology. Bacterial chondronecrosis associated with S aureus has thus been identified as an important cause of leg weakness in these commercial broilers. Lesions suggestive of the condition were visible macroscopically in only 11.1 per cent of the cases subsequently diagnosed by histology and bone histology is therefore required before a diagnosis can be excluded. Angular limb deformities (13.6 per cent) and spondylolisthesis (11.4 per cent) were the most common macroscopic lesions identified as causes of lameness. The overall incidence of tibial dyschondroplasia was similar in both the lame and sound broilers, but severe lesions were found only in lame birds (4.5 per cent). PMID- 9725186 TI - Retrospective survey of allergen immunotherapy in canine atopy. AB - The clinical records of 277 cases of canine atopy treated with specific allergen immunotherapy were reviewed. A good response was defined as control with immunotherapy either alone or with topical agents, a partial response as control with immunotherapy and other systemic agents, and a poor response as no perceived benefit and the immunotherapy discontinued. The mean follow-up period was 29.2 months (range 10 to 85 months). Ninety-one cases (33 per cent) were lost to follow-up or failed to comply with the therapeutic protocol. Of the remaining 186 cases, 40 (21.5 per cent) had a good response to immunotherapy, 74 (39.8 per cent) had a partial response, and 72 (38.7 per cent) had a poor response. Immunotherapy was therefore of long-term benefit in 114 dogs (61.3 per cent). No significant differences in response rates were associated with the breed or sex of the dog, or the age of onset of the disease, or with the type or number of allergens included in a vaccine. Dogs which had clinical signs for more than 61 months before immunotherapy had a significantly poorer response rate (23.5 per cent, P < 0.05). In-house cases had a significantly better response rate (95.2 per cent, P < 0.05) than externally managed cases. PMID- 9725185 TI - Serological and immunohistochemical study of African swine fever in wild boar in Spain. AB - A serological and immunohistochemical study of African swine fever was carried out in wild boar killed in seven municipalities in the north of the province of Cordoba during two hunting seasons (1991-92 and 1992-93), when the area was affected by the disease. Fourteen of 147 wild boar analysed by ELISA and immunoblotting had antibodies to African swine fever virus. The immunohistochemical study revealed that four cases (two seropositive and two seronegative) showed immunoreactivity to the anti-VP73 monoclonal antibody. Two of the VP73+ wild boar had severe generalised haemorrhages consistent with the acute from of the disease, and another had lesions consistent with subacute African swine fever, but none of the remaining 144 animals had gross or microscopic changes suggestive of the disease. These results indicate that wild boar can suffer from African swine fever without showing clinical signs. The disease in wild boar was associated with the disease in domestic pigs. Thus, no African swine fever-positive boar were found either in one municipality with no out-breaks in domestic pigs or in three municipalities with only one outbreak in pigs during the hunting seasons and during the previous year. These results suggest that European wild boar do not play an important role as carriers of the virus of African swine fever. PMID- 9725187 TI - Carriage of trichomonads, Hexamita species and Blastocystis species by adult pheasants. PMID- 9725188 TI - Oestrus synchronisation combined with buserelin administration in beef cattle. PMID- 9725189 TI - Investigations of a rinderpest outbreak in buffaloes in Pakistan. PMID- 9725191 TI - Mastitis in a dairy herd associated with Corynebacterium bovis. PMID- 9725190 TI - Recall of Droplix. PMID- 9725192 TI - Bogus veterinary surgeon. PMID- 9725193 TI - Osteocarcinoma in a Siamese kitten. PMID- 9725194 TI - Cataracts in farmed Atlantic salmon. PMID- 9725195 TI - Rabies in a Zambian bat. PMID- 9725196 TI - Fc gamma RII-B1 regulates the presentation of B cell receptor-bound antigens. AB - Fc gamma receptors (Fc gamma RII) on B lymphocytes negatively regulate B cell receptor (BCR)-dependent activation upon cross-linking of the two receptors. The mechanism reflects the ability of the Fc gamma RII cytoplasmic tail to recruit specific phosphatases that inactivate elements of the BCR-signaling cascade. We now show that cross-linking also blocks the processing and presentation of BCR bound Ag. This occurs because the Fc gamma RII isoform typically expressed by B cells (Fc gamma RII-B1) is incompetent for endocytosis. When cross-linked, Fc gamma RII-B1 acts as a dominant negative inhibitor of BCR endocytosis. In contrast, cross-linking of endocytosis-competent Fc gamma RII isoforms did not inhibit endocytosis or processing of BCR-bound Ag. Thus, Fc gamma RII-B1 acts not only to prevent B cell activation under conditions of Ab excess, but also to prevent clonotypic T cell activation by inhibiting the ability of B cells to generate specific MHC class II-bound TCR ligands. PMID- 9725197 TI - CXCR4 is a functional coreceptor for infection of human macrophages by CXCR4 dependent primary HIV-1 isolates. AB - The identification of HIV-1 coreceptors has provided a molecular basis for the tropism of different HIV-1 strains. CXC chemokine receptor-4 (CXCR4) mediates the entry of both primary and T cell line-adapted (TCLA) syncytia-inducing strains. Although macrophages (M phi) express CXCR4, this coreceptor is assumed to be nonfunctional for HIV-1 infection. We addressed this apparent paradox by infecting human monocyte-derived M phi with primary and TCLA isolates that were rigorously characterized for coreceptor usage and by adding the natural CXCR4 ligand, stem cell differentiation factor-1, to specifically block CXCR4-mediated entry. Our results show that primary HIV-1 isolates that selectively use CXCR4 productively infected both normal and C-C chemokine receptor-5-null M phi. By contrast, M phi supported the entry of CXCR4-dependent TCLA strains with variable efficiency but were not productively infected. Thus, the tropism of HIV isolates results from complex virus/host cell interactions both at the entry and postentry levels. PMID- 9725198 TI - Human anaphylatoxin C4a is a potent agonist of the guinea pig but not the human C3a receptor. AB - The interaction of human anaphylatoxin C4a with the guinea pig (gp) and human (hu) C3a receptors (C3aR) was analyzed using human rC4a, which exhibited C4a specific activity on guinea pig platelets. A gpC3aR of 475 residues with a large second extracellular loop and a peptide sequence approximately 60% identical to the huC3aR was isolated from a genomic DNA library and found to be expressed in guinea pig heart, lung, and spleen. HEK-293 cells cotransfected with this clone, and a cDNA encoding G alpha-16 specifically bound (Kd = 1.6+/-0.7 nM) and responded functionally to C3a with an intracellular calcium mobilization (ED50 = 0.18+/-0.02 nM). Human rC4a weakly bound to both the hu- and gpC3aR (IC50 > 1 microM). However, only HEK-293 cells expressing the gpC3aR responded functionally to rC4a (ED50 = 8.7+/-0.52 nM), while cells expressing the huC3aR did not (c < or = 1 microM). Thus, through an interaction with the C3aR, huC4a may elicit anaphylatoxic effects in guinea pigs but not in man. PMID- 9725199 TI - Dendritic cells require maturation via CD40 to generate protective antitumor immunity. AB - A critical role for CD40/CD154 interactions in the generation of protective cell mediated tumor immunity has been demonstrated previously. Herein, we show that the failure to generate systemic tumor immunity in the absence of CD40/CD154 interactions correlates with an inhibition of Th1-type cytokine production following tumor vaccination. Furthermore, protective antitumor responses can be restored in CD40-deficient mice by the coadministration of CD40+/+ but not CD40-/ dendritic cells (DCs) with tumor Ag, suggesting that CD40 is critical for the maturation and function of DCs in vivo. Finally, we demonstrate that an IL-12 transduced but not a mock-transduced tumor vaccine induces systemic tumor immunity in anti-CD154-treated and CD154-deficient mice. These data suggest that impaired antitumor responses in the absence of CD40/CD154 interactions are the result of a lesion in APC function, namely IL-12 production, and that CD40 plays a critical role in the maturation of DCs in vivo. PMID- 9725200 TI - Constitutive IL-10 production accounts for the high NK sensitivity, low MHC class I expression, and poor transporter associated with antigen processing (TAP)-1/2 function in the prototype NK target YAC-1. AB - Tumor cells that are treated with rIL-10 or transfected with the IL-10 gene show phenotypic changes. These include low but peptide-inducible expression of MHC class I, low sensitivity to specific CTL-mediated lysis, and increased NK sensitivity. In vitro-established mouse tumor lines were screened for IL-10 expression and production, and a large proportion of plasmocytomas or T cell lymphomas were found to produce IL-10. Since one of these lines was the prototype NK target cell YAC-1, we investigated whether the high IL-10 production of this cell line was related to its high NK sensitivity and its defects in MHC class I expression. The decrease in H-2 expression following the in vitro culture of in vivo-passaged YAC-1 cells was accompanied by a gradual increase in IL-10 production, whereas the reverse was found when passing in vitro-grown YAC-1 in vivo as an ascites tumor in syngenic mice. In addition, differences in YAC-1 MHC class I expression correlated with alterations in the functional activity of TAP 1/2 proteins. YAC-1 cells that were transduced with a retroviral IL-10 antisense construct (Y-IL-10 AS) only produced about half of the IL-10 that was produced by YAC-1 transduced with the control construct (Y-IL-10 Mock). Relative to Y-IL-10 Mock cells, the expression of H-2 on Y-IL-10 AS cells was markedly increased, and NK sensitivity was decreased. These data argue for a mechanism wherein IL-10 production is causally related to the low H-2 expression, decreased TAP function, and high NK sensitivity of YAC-1 cells. PMID- 9725201 TI - MHC class II transport from lysosomal compartments to the cell surface is determined by stable peptide binding, but not by the cytosolic domains of the alpha- and beta-chains. AB - Inside APCs, MHC class II molecules associate with antigenic peptides before reaching the cell surface. This association takes place in compartments of the endocytic pathway, more related to endosomes or lysosomes depending on the cell type. Here, we compared MHC class II transport from endosomal vs lysosomal compartments to the plasma membrane. We show that transport of MHC class II molecules to the cell surface does not depend on the cytosolic domains of the alpha- and beta-chains. In contrast, the stability of the alphabeta-peptide complexes determined the efficiency of transport to the cell surface from lysosomal, but not from endosomal, compartments. In murine B lymphoma cells, SDS unstable and -stable complexes were transported to the cell surface at almost similar rates, whereas after lysosomal relocalization or in a cell line in which MHC class II molecules normally accumulate in lysosomal compartments, stable complexes were preferentially addressed to the cell surface. Our results suggest that when peptide loading occurs in lysosomal compartments, selective retention and lysosomal degradation of unstable dimers result in the expression of highly stable MHC class II-peptide complexes at the APC surface. PMID- 9725202 TI - Beta-galactoside-binding protein (beta GBP) alters the cell cycle, up-regulates expression of the alpha- and beta-chains of the IFN-gamma receptor, and triggers IFN-gamma-mediated apoptosis of activated human T lymphocytes. AB - In this paper, the effects of beta-galactoside binding protein (beta GBP), the LGALS1 gene product, on the cell cycle progression and expansion of activated human T lymphocytes were studied. Beta GBP drastically inhibits the IL-2 induced proliferation of PHA-activated T lymphocytes as well as the IL-2 independent proliferation of malignant T lymphocytes by arresting them in the S and G2/M phases of the cell cycle. In addition, beta GBP up-regulates the expression of both the alpha- and the beta-chains of the IFN-gamma R on activated T lymphocyte membrane. None of these effects depend on sugar binding: saturating amounts of lactose do not affect the cell cycle block nor IFN-gamma R up-modulation. The increased expression of both chains renders beta GBP-treated T lymphoblasts sensitive to IFN-y-induced apoptosis. Taken as a whole, these findings suggest that beta GBP plays an important immunoregulatory role by switching off T lymphocyte effector functions. They also provide the first evidence of up modulation of IFN-gamma R expression on T lymphocytes by a negative cell growth regulator. PMID- 9725204 TI - Autoimmunity without diabetes in transgenic mice expressing beta cell-specific CD86, but not CD80: parameters that trigger progression to diabetes. AB - To define more clearly the roles of CD80 (RIP-CD80) and CD86 (RIP-CD86) in the activation of autoreactive T cells in vivo, we generated transgenic mice expressing either or both costimulatory molecules on the beta cells of the pancreas. While RIP-CD80 mice do not show any sign of autoimmunity, at the age of 7 mo RIP-CD86 transgenic mice develop a lymphoid infiltrate with both IFN-gamma- and IL-4-positive cells in the vicinity of the islets; these mice, however, never progress to diabetes. This fundamental difference in the ability of CD80 and CD86 to activate self-reactive T cells in vivo is, however, obliterated when the level of TCR signaling is increased by either TNF-alpha or transgenic MHC class II expression. These results support the suggestion that CD80 and CD86 mainly differ at the level of the intensity of the signals they deliver. PMID- 9725203 TI - Switch of CD4+ T cell differentiation from Th2 to Th1 by treatment with cathepsin B inhibitor in experimental leishmaniasis. AB - When activated, CD4+ T helper cells differentiate functionally into one of two subsets, Th1 or Th2. Before the Th differentiation, Ags must be processed into peptide epitopes and presented to CD4+ T cells in association with MHC class II molecules. However, the proteases responsible for this Ag processing have not been well defined. When BALB/c mice susceptible to infection with Leishmania major were treated with a specific inhibitor (CA074) of cathepsin B, a lysosomal cysteine protease that digests exogenous antigenic proteins, those mice acquired resistance against infection with L. major and showed the shift of immune responses from Th2 to Th1; that is, they produced specific IgG2a Ab and generated IFN-gamma in contrast to untreated and infected mice that produced IgG1 and IgE and generated IL-4. CA074 interfered with the digestion of L. major Ags with lysosomal enzymes in vivo as well as in vitro. However, this inhibitor did not show any direct influence on the growth of L. major and the functions of T cells stimulated with anti-CD3 Ab. These findings indicate that cathepsin B inhibitor could switch CD4+ T cell differentiation from Th2 to Th1, suggesting that the alteration in Ag processing modulates the polarity of Th differentiation. PMID- 9725206 TI - Gallium arsenide modulates proteolytic cathepsin activities and antigen processing by macrophages. AB - Gallium arsenide (GaAs) is a semiconductor utilized in the electronics industry. Chemical exposure of animals causes a local inflammatory reaction, but systemic immunosuppression. Mice were administered i.p. 200 mg/kg GaAs crystals or latex beads, or vehicle. Five days after exposure, splenic macrophages were defective, whereas thioglycolate-elicited peritoneal macrophages (PEC) were more efficient in processing the Ag, pigeon cytochrome c, than vehicle control macrophages. Various aspects of the MHC class II Ag-processing pathway were examined. Both macrophage populations normally presented a peptide fragment to the CD4+ T cells. Surface MHC class II expression on the PEC was up-regulated, but splenic cells had normal MHC class II expression. PEC had elevated levels of glutathione and cysteine, major physiologic reducing thiols. However, the cysteine content of splenic macrophages was diminished. Proteolytic activities of aspartyl cathepsin D, and thiol cathepsins B and L were decreased significantly in splenic macrophages. On the other hand, thiol cathepsin activities were increased selectively in PEC. Latex bead-exposed PEC were not more potent APC, and their thiol cathepsin activities were unchanged, indicating that phagocytosis and nonspecific irritation were not responsible. The phenotype of PEC directly exposed to GaAs mirrored cytokine-activated macrophages, in contrast to splenic macrophages from a distant site. Therefore, GaAs exposure differentially modulated cathepsin activities in splenic macrophages and PEC, which correlated with their Ag-processing efficiency. Perhaps such distinct alterations may contribute to the local inflammation and systemic immunotoxicity caused by chemical exposure. PMID- 9725205 TI - Quiescence-inducing and antiapoptotic activities of IL-15 enhance secondary CD4+ T cell responsiveness to antigen. AB - IL-15 shows functional redundancy with IL-2 due to its usage of the beta and gamma c subunit of the IL-2R. Yet, the requirement of IL-15 for an IL-15R alpha chain for high affinity interaction and the separate cellular sources of IL-2 and IL-15 suggest divergent activities for both cytokines. We compared the growth inducing and proapoptotic or antiapoptotic activities of IL-15 and IL-2 on mature CD4+ T lymphocytes in the presence or absence of TCR occupancy. We found that the nature of IL-15 activity was critically dependent on the activation status of the T cells. In the absence of TCR triggering, IL-15 did not exert the growth factor activity of IL-2, but induced a quiescent phenotype, characterized by maintenance of the cells in the G0/G1 phase of the cell cycle and down-regulation of CD25, CD71, and CD95 expression. In the presence of appropriate TCR engagement, the IL 15-induced quiescent T cells were resistant against TCR-induced cell death and proliferated strongly. IL-2-treated cells, on the contrary, were sensitized to cell death, resulting in a negative feedback on cellular expansion and weak proliferative responsiveness. Consecutive action of IL-15 during the distinct phases of an in vitro immune response markedly increased the cell output of a second antigenic stimulation, as compared with IL-2. These results imply that during immune reactivity in vivo, IL-15 may take over from the transiently available IL-2 the role of survival factor but not of growth factor, hence promoting the long term maintenance of resting, Ag-experienced CD4+ T cells. PMID- 9725207 TI - Human endothelial cells effectively costimulate cytokine production by, but not differentiation of, naive CD4+ T cells. AB - We compared costimulatory signals provided by human endothelial cells (ECs) to those provided by conventional bone marrow-derived APCs, i.e., peripheral blood adherent mononuclear cells (PBAMCs), by measuring their effects on cytokine production by naive or memory CD4+ T cells stimulated by PHA. In these assays, ECs effectively costimulate secretion of IL-2, IFN-gamma, and IL-4 from both naive and memory CD4+ T cells, quantified by ELISA or intracellular cytokine staining. ECs, which lack B7 molecules, use predominantly leukocyte-function associated Ag 3 (LFA-3) to provide costimulation. ECs are comparable to or better than PBAMCs, which use both the LFA-3 and B7 molecules, at costimulating IL-2 and IL-4 production. ECs are less effective than PBAMCs at costimulating IFN-gamma production by naive T cells. ECs do not secrete IL-12, and addition of exogenous IL-12 enables ECs to costimulate IFN-gamma at a level comparable to that observed with PBAMCs. ECs do not promote differentiation of naive T cells to Th1-like cells, whereas PBAMCs do. Again, addition of exogenous IL-12 enables ECs to do so. Transfection of ECs to express B7-1 or B7-2 is less effective than IL-12 supplementation for restoring these responses. These experiments suggest that a deficiency in costimulation due to lack of B7 molecule expression does not fully explain the inability of ECs to activate resting naive CD4+ T cells. PMID- 9725208 TI - Peripheral immune tolerance blocks clonal expansion but fails to prevent the differentiation of Th1 cells. AB - Clonal anergy in Ag-specific CD4+ T cells is shown in these experiments to inhibit IL-2 production and clonal expansion in vivo. We also demonstrate that the defect in IL-2 gene inducibility can be achieved in both naive and Th1-like memory T cells when repeatedly exposed to aqueous peptide Ag. Nevertheless, this induction of clonal anergy did not interfere with the capacity of naive T cells to differentiate into Th1-like effector cells, nor did it prevent such helper cells from participating in T-dependent IgG2a anti-hapten responses and delayed type hypersensitivity reactions. Thus, clonal anergy can contribute to the development of Ag-specific immune tolerance by limiting the size of a Th cell population, but not by disrupting its effector function. PMID- 9725209 TI - Vaccination with BV8S2 protein amplifies TCR-specific regulation and protection against experimental autoimmune encephalomyelitis in TCR BV8S2 transgenic mice. AB - TCR determinants overexpressed by autopathogenic Th1 cells can naturally induce a second set of TCR-specific regulatory T cells. We addressed the question of whether immune regulation could be induced naturally in a genetically restricted model in which a major portion of TCR-specific regulatory T cells expressed the same target TCR BV8S2 chain as the pathogenic T cells specific for myelin basic protein (MBP). We found vigorous T cell responses to BV8S2 determinants in naive mice that could be further potentiated by vaccination with heterologous BV8S2 proteins, resulting in the selective inhibition of MBP-specific Th1 cells and protection against experimental encephalomyelitis. Moreover, coculture with BV8S2 specific T cells or their supernatants reduced proliferation, IFN-gamma secretion, and encephalitogenic activity of MBP-specific T cells. These results suggest that immune regulation occurs through a nondeletional cytokine-driven suppressive mechanism. PMID- 9725210 TI - Tumor-infiltrating lymphocytes exhibiting high ex vivo cytolytic activity fail to prevent murine melanoma tumor growth in vivo. AB - The identification of tumor-associated Ags recognized by CD8+ CTL and prevention of tumor outgrowth by adoptive transfer of these CTL demonstrates that CD8+ T cells play a major role in antitumor immunity. We have generated B16.F10 melanoma cells that express the glycoprotein epitope amino acid 33-41 (GP33) of the lymphocytic choriomeningitis virus (LCMV) to examine antitumor CD8+ T cell response in C57BL/6 mice immune to LCMV and in mice transgenic for the LCMV GP33 specific P14 TCR (P14 TCR mice). We find that B16.F10GP33 tumor cells grew in syngeneic C57BL/6 mice without inducing T cell tolerance. LCMV infection or adoptive transfer of LCMV-specific effector T cells delayed but did not prevent growth of preestablished tumors in these mice. However, B16.F10GP33 tumor cells were rejected in mice immune to LCMV and in mice treated with LCMV-specific effector T cells on the same day as the tumor. Surprisingly, B16.F10GP33 tumor cells grew in P14 TCR transgenic mice despite an abundance of tumor-associated Ag specific CD8+ T cells. In these mice, freshly isolated tumor-infiltrating lymphocytes exhibited an activated phenotype and displayed high GP33-specific cytolytic activity when assessed ex vivo. Thus, B16.F10GP33 melanoma cells are able to initiate, but not to sustain, a GP33-specific CTL response sufficient to clear the tumor enduringly. PMID- 9725211 TI - TNF-related apoptosis-inducing ligand (TRAIL) induces apoptosis in Fas ligand resistant melanoma cells and mediates CD4 T cell killing of target cells. AB - We have previously shown that melanoma cells were resistant to apoptosis induced by TNF family members Fas ligand (FasL), TNF-alpha, and CD40L. FasL also was not involved in CD4 T cell-mediated killing of melanoma cells. In the present study, we have tested melanoma cells for their susceptibility to apoptosis induced by human TNF-related apoptosis-inducing ligand (TRAIL) and the ability of a mAb against TRAIL to inhibit apoptosis and CD4 CTL-mediated killing of melanoma and Jurkat target cells. The results show that TRAIL-induced apoptosis in cells from 7 of 10 melanoma cell lines tested as well as in Jurkat T cells. Susceptibility to apoptosis was increased in some of the cell lines by treatment with cyclohexamide or actinomycin D. The melanoma cells were resistant to apoptosis induced by FasL, TNF-alpha, and CD40L. mAb M180 against TRAIL inhibited apoptosis induced by TRAIL. It was also found to inhibit CD4 CTL-mediated killing of Jurkat T cells as well as autologous and allogeneic melanoma cells. The degree of inhibition produced by the mAb varied between different clones of CTL and according to the susceptibility of the target cells to TRAIL-induced apoptosis. These results suggest that TRAIL is an important mediator of cell death induced by CTL and may have an important therapeutic role against human melanoma. PMID- 9725212 TI - Protein kinase C regulates Fas (CD95/APO-1) expression. AB - Fas (CD95/APO-1) is a transmembrane protein of the TNF/neuron growth factor receptor family. Ligation of Fas by specific Abs or Fas ligand (FasL/CD95 ligand) induces rapid apoptotic cell death in a variety of cell types. Despite progress in understanding the death signals transduced from Fas, very little is known with regard to the mechanisms by which Fas expression is regulated. Using our previously established murine T cell hybridoma model A1.1, we show that specific protein kinase C (PKC) inhibitors could block activation-induced Fas expression and apoptosis. The activation of PKC with PMA or 1-oleoyl-2-acetyl-sn-glycerol could mimic the TCR signal by inducing the expression of Fas but not FasL. PKC dependent Fas expression was also observed in several murine and human tumor cell lines. Since the inhibition of Ca2+ redistribution by an inhibitor of intracellular Ca2+ mobilization, 8-(diethylamino)-octyl-3,4,5-trimethoxybenzoate hydrochloride, inhibited TCR-induced FasL but not Fas, the expression of Fas appears to be independent of Ca2+ mobilization. Significantly, expression of the newly identified Fas-regulatory gene, TDAG51, was found to be dependent upon the activity of PKC. PKC activation only induced Fas expression in cells expressing wild-type TDAG51. Thus, Fas expression is likely mediated by PKC through TDAG51. PMID- 9725213 TI - Differential requirements of CD45 protein tyrosine phosphatase for cytolytic activities and intrathymic and extrathymic development of intestinal intraepithelial lymphocytes. AB - CD45 is a transmembrane protein tyrosine phosphatase essential for Ag receptor mediated signaling in both T and B cells. In this study we investigated roles of CD45 in development and cytolytic activities of murine intestinal intraepithelial lymphocytes (i-IEL) using CD45 exon 6 knockout (CD45-/-) mice. Interestingly, the total cell number of i-IEL was significantly reduced in CD45-/-mice during aging (10-20 wk of age), whereas the i-IEL number was normally increased in the wild type littermates. Especially, the number of gamma(delta)TCR+ i-IEL decreased markedly in CD45-/- mice during aging. The i-IEL in CD45-/- mice were more susceptible to in vitro spontaneous apoptosis than the normal i-IEL, implying that CD45 is required for maintenance of the cellularity of i-IEL. Results from in vivo analyses of the extrathymic and intrathymic development of i-IEL suggested that CD45-mediated signaling is required for the intrathymic, but not the extrathymic, development of i-IEL. Moreover, the whole i-IEL from CD45-/- mice showed a significantly reduced cytolytic activity, and the residual cytolytic activity was completely diminished by depleting CD45+ i-IEL, suggesting that CD45 is indispensable for the TCR-mediated cytolytic activity of i-IEL. Furthermore, we found differential contributions of CD45 and p56lck to development and induction of cytolytic activities of i-IEL. PMID- 9725214 TI - IL-18 augments perforin-dependent cytotoxicity of liver NK-T cells. AB - The liver contains abundant cytotoxic cells, including NK-T cells, NK cells, and CTLs. However, the regulation of this cytotoxicity is not fully understood. In this study, we investigated the effect of a recently described cytokine, IL-18, which is present in large quantities in the liver, on the cytotoxicity of intrahepatic lymphocyte subpopulations. This effect of IL-18 was assessed by assaying the in vitro cytotoxicity of purified NK-T, NK, and T cells against a CD95- and perforin-sensitive T cell line, Jurkat. The results show that IL-18 enhances the killing activity of liver NK-T cells by a CD95-independent, perforin dependent pathway. IL-18 also augments liver NK cell activity, but the exact mechanisms of this killing remain to be elucidated. Finally, the augmentation of the killing activities of liver NK-T and NK cells by IL-18 is not due to soluble TNF-alpha, because none of these cell populations had detectable TNF-alpha production. PMID- 9725215 TI - Critical role of IL-12 in dendritic cell-induced differentiation of naive B lymphocytes. AB - Dendritic cells (DC) are potent APCs initiating immune responses. In a previous report, we demonstrated that DC directly enhance both proliferation and differentiation of CD40-activated naive and memory B cells. The present study deciphers the molecular mechanisms involved in DC-dependent regulation of B cell responses. Herein, we have identified IL-12 as the mandatory molecule secreted by CD40-activated DC that promote the differentiation of naive B cells into plasma cells secreting high levels of IgM. In fact, IL-12 synergizes with soluble IL-6R alpha-chain (sgp80), produced by DC, to drive naive B cell differentiation. IL-12 is critical for the differentiation of naive B cells into IgM plasma cells, whereas IL-6R signaling mainly promotes Ig secretion by already differentiated B cells. The differentiation of naive B cells in cocultures of B cells, T cells, and DC is IL-12 dependent, definitely demonstrating that the role of DC in humoral responses is not confined to the activation of T cells and further extending the physiologic relevance of DC/B cell interaction. Finally, this study also identifies differential requirements for DC-dependent naive and memory B cell differentiation, the latter being IL-12 independent. Altogether these results indicate that, in addition to prime T cells toward Thl development, DC, through the production of IL-12, may also directly signal naive B cell during the initiation of the immune response. PMID- 9725216 TI - Substance P (neurokinin-1) receptor is a marker of human mucosal but not peripheral mononuclear cells: molecular quantitation and localization. AB - Reciprocal communication between the immune system and the neuroendocrine system is mediated via a common chemical language of shared ligands and receptors. The neuropeptide substance P (SP) has been implicated as a mediator of immunomodulation. The evidence for substance P receptors on human lymphocytes is, however, controversial. The aims of the present study are to investigate substance P receptor (SPR) expression in human peripheral and mucosal mononuclear cells and to identify cellular sites of expression in human colonic mucosa. Using reverse-transcriptase PCR, we demonstrate that PBMC isolations are negative for SPR mRNA expression, whereas lamina propria mononuclear cell (LPMC) isolations express on average eight SPR mRNA transcripts per cell. In situ hybridization performed on surgically resected colonic tissue confirms the expression of SPR mRNA in LPMC in vivo. SPR mRNA signal was detected in LPMC, lymphoid follicles, and epithelium. The complementary technique of immunohistochemistry gave a similar distribution of SPR expression that colocalized with CD45 immunoreactivity. Dual-fluorochrome flow cytometry revealed SPR expression by CD4, CD45RO, CD45RA, CD8, CD19, and CD14 LPMC subsets, but not PBMC. Our findings suggest that SPR expression is distinctive of human colonic mucosal mononuclear cells and support a direct role for SP in mucosal immunomodulation. PMID- 9725217 TI - On histocompatibility barriers, Th1 to Th2 immune deviation, and the nature of the allograft responses. AB - In the present study, we have sought to determine the basis for the frequent failure of Th1 to Th2 immune deviation to blunt the severity of allograft rejection, as such immune deviation has proven highly effective in the treatment of several T cell-dependent autoimmune states. Our study demonstrates that treating islet allograft recipient mice with anti-IL-12 mAb is highly effective in producing Th1 to Th2 immune deviation in several model systems (i.e., fully MHC, partially MHC, or multiple minor Ag barriers). Nevertheless, anti-IL-12 failed to prolong the engraftment of fully MHC-mismatched islet allografts. However, anti-IL-12-treated recipients carrying MHC-matched but multiple minor Ag mismatched allografts experienced prolonged engraftment; allograft tolerance was frequently achieved in the DBA/2J (H-2d) to BALB/c (H-2d) strain combination. In another model, in which the host response was dominated by CD4+ T cells responding to donor allopeptides presented upon host APCs in the context of self MHC class II molecules, anti-IL-12 treatment proved to be extremely potent. Thus, Th1 to Th2 immune deviation produces prolonged engraftment as compared with recipients of MHC-mismatched allografts when rejection is dependent upon indirectly presented allogeneic peptides and a reduced mass of responding alloreactive T cells. PMID- 9725218 TI - Maturation of B cell precursors is impaired in thymic-deprived nude and old mice. AB - We have previously reported that bone marrow B cell precursors from thymic deprived nude and old mice express less recombination-activating gene-1 (RAG-1) mRNA than they do in young euthymic mice. We now report that both nude and old mice have decreased bone marrow pre-B cells and that fewer pre-B cells express RAG protein. This combination of events appears to be the basis for the lower level of bone marrow RAG mRNA in thymic-deprived mice. A link between thymic function and B cell development was suggested by the similar kinetics of thymic involution and of declining bone marrow RAG-1 gene expression during aging. Support for this hypothesis was obtained by demonstrating that injection of supernatant medium from activated CD8+ but not CD4+ young T cells from mice increases RAG mRNA, RAG protein, and the number of bone marrow pre-B cells in nude and old mice. Furthermore, in vivo CD8+ T cells also regulate bone marrow RAG gene expression. Thus, mice deficient in CD8+ T cells expressed levels of RAG 1 mRNA in their bone marrow that were only 10% of those observed in normal or CD4+ T cell-deficient mice. IL-16 was detected in the supernatant medium from activated T cell cultures, and injection of nanogram quantities of recombinant IL 16 (rIL-16) into nude or old mice increased the levels of RAG mRNA in bone marrow B cell precursors and the number of bone marrow pre-B cells. We conclude that the impaired development of B cells within the bone marrow of thymic-deprived nude and old mice can be reversed, at least in part, by the administration of rIL-16. PMID- 9725219 TI - Distinct stage-specific cis-active transcriptional mechanisms control expression of T cell coreceptor CD8 alpha at double- and single-positive stages of thymic development. AB - Developing thymocytes that give rise to CD8+ (cytotoxic) and CD4+ (helper) alpha beta-TCR T lymphocytes go through progressive stages of expression of coreceptors CD8 and CD4 from being negative for both (the double-negative stage), to coexpressing both (the double-positive (DP) stage), to a mutually exclusive sublineage-specific expression of one or the other (the single-positive (SP) stage). To delineate the mechanisms underlying regulation of CD8 during these developmental transitions, we have examined expression of a series of mouse CD8 alpha gene constructs in developing T cells of conventional and CD8 alpha "knock out" transgenic mice. Our results indicate that cis-active transcriptional control sequences essential for stage- and sublineage-specific expression lie within a 5' 40-kb segment of the CD8 locus, approximately 12 kb upstream of the CD8 alpha gene. Studies to characterize and sublocalize these cis sequences showed that a 17-kb 5' subfragment is able to direct expression of the CD8 alpha gene up to the CD3intermediate DP stage but not in more mature DP or SP cells. These results indicate that stage-specific expression of CD8 alpha in developing T cells is mediated by the differential activity of multiple functionally distinct cis-active transcriptional control mechanisms. It will be important to determine the relationship of "switching" between these cis mechanisms and selection. PMID- 9725220 TI - Transcription of a minimal promoter from the NF-IL6 gene is regulated by CREB/ATF and SP1 proteins in U937 promonocytic cells. AB - NF-IL6 is an important transcriptional regulator of genes induced in activated monocytes/macrophages, and NF-IL6 is the only CCAAT/enhancer-binding protein (C/EBP) family member whose steady-state mRNA levels increase upon activation of monocytes (1). We show that increased transcription of the NF-IL6 gene is responsible, at least in part, for induction of NF-IL6 mRNA following activation of U937 promonocytic cells. We have identified a 104-bp minimal promoter region of the NF-IL6 gene that is sufficient for basal and activation-dependent induction of transcription in U937 cells. This region contains binding sites for the cAMP response element-binding protein/activation transcription factor (CREB/ATF) and Sp1 families of transcription factors. Each site is functionally important and contributes independently to transcription of the NF-IL6 gene in U937 cells. PMID- 9725221 TI - The role of enhancer A in the locus-specific transactivation of classical and nonclassical HLA class I genes by nuclear factor kappa B. AB - HLA class I expression is tightly controlled at the transcriptional level by several conserved regulatory elements in the proximal promoter region. In this study, the two putative kappa B motifs of enhancer A (kappa B1 and kappa B2) of the classical and nonclassical HLA class I genes were investigated for their binding properties of transcription factors and tested for their contribution to the NF-kappa B-induced route of transactivation. It was shown that NF-kappa B induced transactivation through enhancer A is most important for the HLA-A locus, which contains two NF-kappa B binding sites. Although the enhancer A of HLA-B contains only one NF-kappa B binding site (kappa B1), there was still a moderate transactivation by NF-kappa B. Since HLA-F, which also possesses one NF-kappa B binding site but lacks protein binding to its KB2 site, was not transactivated by NF-kappa B, the NF-kappa B-mediated transactivation through the kappa B1 motif in HLA-B is most probably facilitated by binding of the transcription factor Spl to the upstream kappa B2 site. Thus, transcriptional regulation of HLA class I genes by NF-kappa B is restricted to the HLA-A and HLA-B loci. PMID- 9725222 TI - Rheumatoid factor specificity of a VH3-encoded antibody is dependent on the heavy chain CDR3 region and is independent of protein A binding. AB - Rheumatoid factors (RF) recognize conformational determinants located within the Fc portion of IgG. By analyzing a panel of monoclonal rheumatoid arthritis (RA) derived RFs, we previously demonstrated that the somatically generated light chain complementarity-determining region 3 (CDR3) contributes to RF specificity. We have now generated a panel of heavy chain mutants of the B'20 Ab, a high affinity RA-derived IgM RF. B'20 also binds avidly to protein A and weakly to ssDNA and tetanus toxoid. B9601, a RF negative Ab that is highly homologous to B'20 but does not bind any of the Ags tested, and RC1, a low affinity polyreactive RF, were used to generate heavy chain mutants with framework (FR) and CDR switches. The mutated heavy chains were cotransfected into a myeloma cell line with the germline counterpart of the B'20 light chain, and the expressed Ig tested for antigenic specificity. We show that both RF specificity and polyreactivity of B'20 is dependent on its unique heavy chain CDR3 region. Replacement with a B9601 CDR3 shortened to the same length as the B'20 CDR3, and with only 5 amino acid differences, did not restore Fc binding. Conversely, absence of protein A binding of B9601 is due to the presence of a serine residue at position 82a in the B9601 heavy chain FR3 region. Together, our data suggest that Ig gene recombination events can generate B cells with autoantibody specificities in the preimmune repertoire. Abnormal release, activation, expansion, or mutation of such cells might all contribute to the generation of a high titer RF response in patients with RA. PMID- 9725223 TI - Asymmetric contribution to Ig repertoire diversity by V kappa exons: differences in the utilization of V kappa 10 exons. AB - The mouse has approximately 140 germline V kappa genes, and functional V kappa exons are expressed at roughly equivalent levels in the preimmune repertoire. We have examined the expression of individual members of the V kappa 10 family. V kappa 10A and V kappa 10B genes have been utilized in numerous hybridomas and myelomas, while V kappa 10C has not. In this study, we have cloned the V kappa 10C gene and shown that it is structurally functional, has the expected promoter elements and recombination signal sequences, and that it is capable of recombination. V kappa 10C mRNA, however, is present at levels at least 1000-fold lower than V kappa 10A and V kappa 10B in adult spleens. While there are no sequence differences in the octamer or TATA box between V kappa 10C and V kappa 10A, there are three nucleotide changes in the promoter region. These promoters equally drive the expression of a reporter gene in B cells or plasma cells, but the V kappa 10A promoter is able to drive expression in pre-B cell lines significantly better than the V kappa 10C promoter (p < 0.05). V kappa 10C rearrangements can be detected in bone marrow and splenic DNA. Therefore, the lack of V kappa 10C expression may reflect the inability of V kappa 10C rearranged cells to undergo positive or negative selection. Our results suggest that the available Ab repertoire is shaped not only by the number of structurally functional genes, but also by the ability of assembled genes to be expressed at critical points during B cell maturation. PMID- 9725224 TI - Role of conserved glycosylation site unique to murine class I MHC in recognition by Ly-49 NK cell receptor. AB - The recognition of class I MHC molecules on target cells by the Ly-49 family of receptors regulates NK cytotoxicity. Previous studies have suggested that carbohydrates are involved in the recognition of class I MHC by Ly-49, although their precise role remains unclear. Here, we examined the role of asparagine linked carbohydrates of the murine class I MHC in the binding to Ly-49A and Ly 49C. We have generated H-2Dd mutants that lack the highly conserved glycosylation sites at amino acid residues 86 in the alpha1 domain and 176 in the alpha2 domain, respectively. These mutant Dd cDNAs were transfected into leukemic cell lines, and the binding of the transfected cells to COS cells expressing Ly-49A or Ly-49C, as well as their susceptibility to lysis by Ly-49A+ NK cells, was examined. Only the mutation of the alpha2 domain glycosylation site significantly reduced the binding of Dd to Ly-49A and Ly-49C. Cells expressing Dd with the mutation at this site were partially resistant to killing by Ly-49A+ NK cells. These results suggest that, while carbohydrates linked to residue 176 seem to function as a part of the ligand structure for the Ly-49 family of NK receptors, there are additional structural features involved in this recognition. This glycosylation site is highly conserved among murine class I MHC but is not found among those of other species, suggesting that its role is unique to the murine immune system. It further suggests that murine class I MHC and Ly-49 gene families may have evolved in concert. PMID- 9725225 TI - The assembly and stability of MHC class II-(alpha beta)2 superdimers. AB - X-ray crystallography of several MHC class II molecules revealed a structure described as a dimer of heterodimers, or a superdimer. This discovery led to the hypothesis that MHC class II molecules may interact with the TCR and CD4 as an (alpha beta)2 superdimer, potentially providing more stable and stimulatory interactions than can be provided by the simple alpha beta heterodimer alone. In this study, using chemical cross-linking, we provide evidence for the existence of the superdimers surface of B cells. We further characterize the superdimers and demonstrate that in lysates of B cells, I-Ek dimers and superdimers are derived from the same population of I-Ek molecules. Purified, I-Ek molecules in solution also exist as a mixture of 60-kDa dimers and 120-kDa superdimers, indicating that I-Ek has an intrinsic ability to form 120-kDa complexes in the absence of other cellular components. Peptide mapping showed that the alpha beta and (alpha beta)2 complexes are closely related and that the superdimers do not contain additional polypeptides not present in the dimers. The (alpha beta)2 complex displays thermal and pH stability similar to that of the alpha beta complex, both being denatured by SDS at temperatures above 50 degrees C and at a pH below 5. These data support the model that MHC class II has an intrinsic ability to assume the (alpha beta)2 superdimeric conformation, which may be important for interactions with the TCR and CD4 coreceptor. PMID- 9725226 TI - The murine IL-13 receptor alpha 2: molecular cloning, characterization, and comparison with murine IL-13 receptor alpha 1. AB - Two components of a receptor complex for IL-13, the IL-4R and a low affinity IL 13-binding chain, IL-13R alpha 1, have been cloned in mice and humans. An additional high affinity binding chain for IL-13, IL-13R alpha 2, has been described in humans. We isolated a cDNA from the thymus that encodes the murine orthologue of the human IL-13R alpha 2. The predicted protein sequence of murine IL-13R alpha 2 (mIL-13R alpha 2) has 59% overall identity to human IL-13R alpha 2 and is closely related to the murine low affinity IL-13-binding subunit, IL-13R alpha 1. The genes for both mIL-13-binding chains map to the X chromosome. A specific interaction between mIL-13R alpha 2.Fc protein and IL-13 was demonstrated by surface plasmon resonance using a BIACORE instrument. Ba/F3 cells that were transfected with mIL-13R alpha 2 expressed 5000 molecules per cell and bound IL-13 with a single Kd of 0.5 to 1.2 nM. However, these cells did not proliferate in response to IL-13, and the IL-4 dose response was unaffected by high concentrations of IL-13. In contrast, the expression of mIL-13R alpha 1 by Ba/F3 cells resulted in a sensitive proliferative response to IL-13. Consistent with its lower affinity for IL-13, IL-13R alpha 1.Fc was 100-fold less effective than IL-13R alpha 2.Fc in neutralizing IL-13 in vitro. These results show that mIL-13R alpha 2 and mIL-13R alpha 1 are not functionally equivalent and predict distinct roles for each polypeptide in IL-13R complex formation and in the modulation of IL-13 signal transduction. PMID- 9725228 TI - Protection from lymphoma cell metastasis in ICAM-1 mutant mice: a posthoming event. AB - It has been hypothesized that the intercellular adhesion receptors used by normal cells could also be operative in the spreading of circulating malignant cells to target organs. In the present work, we show that genetic ablation of the ICAM-1 gene confers resistance to T cell lymphoma metastasis. Following i.v. inoculation of LFA-1-expressing malignant T lymphoma cells, we found that ICAM-1-deficient mice were almost completely resistant to the development of lymphoid malignancy compared with wild-type control mice that developed lymphoid tumors in the kidneys, spleen, and liver. Histologic examinations confirmed that ICAM-1 deficient mice, in contrast to wild-type mice, had no evidence of lymphoid infiltration in these organs. The effect of ICAM-1 on T cell lymphoma metastasis was observed in two distinct strains of ICAM-1-deficient animals. Nonetheless, lymphoma cells migrated with the same efficiency to target organs in both normal and ICAM-1-deficient mice, indicating not only that ICAM-1 expression by the host is essential in lymphoma metastasis, but also that this is so at stages subsequent to homing and extravasation into target organs. These results point to posthoming events as a focus of future investigation on the control of metastasis mediated by ICAM-1. PMID- 9725227 TI - A plasmid encoding murine granulocyte-macrophage colony-stimulating factor increases protection conferred by a malaria DNA vaccine. AB - Using the murine parasite Plasmodium yoelii (Py) as a model for malaria vaccine development, we have previously shown that a DNA plasmid encoding the Py circumsporozoite protein (PyCSP) can protect mice against sporozoite infection. We now report that mixing a new plasmid PyCSP1012 with a plasmid encoding murine granulocyte-macrophage colony-stimulating factor (GM-CSF) increases protection against malaria, and we have characterized in detail the increased immune responses due to GM-CSF. PyCSP1012 plasmid alone protected 28% of mice, and protection increased to 58% when GM-CSF was added (p < 0.0001). GM-CSF plasmid alone did not protect, and control plasmid expressing inactive GM-CSF did not enhance protection. GM-CSF plasmid increased Abs to PyCSP of IgG1, IgG2a, and IgG2b isotypes, but not IgG3 or IgM. IFN-gamma responses of CD8+ T cells to the PyCSP 280-288 amino acid epitope increased but CTL activity did not change. The most dramatic changes after adding GM-CSF plasmid were increases in Ag-specific IL-2 production and CD4+ T cell proliferation. We hypothesize that GM-CSF may act on dendritic cells to enhance presentation of the PyCSP Ag, with enhanced IL-2 production and CD4+ T cell activation driving the increases in Abs and CD8+ T cell function. Recombinant GM-CSF is already used in humans for medical purposes, and GM-CSF protein or plasmids may be useful as enhancers of DNA vaccines. PMID- 9725229 TI - Bacterial surface proteins recognized by CD4+ T cells during murine infection with Listeria monocytogenes. AB - Optimal immunity to the Gram-positive pathogen Listeria monocytogenes (LM) requires both CD8+ and CD4+ antigen-specific T cell responses. Understanding how CD4+ T cells function in an immune response to LM and how bacterial proteins are processed to peptide/MHC class II complexes in infected cells requires identification of these proteins. Using LacZ-inducible, LM-specific CD4+ T cells as probes, we identified two immunogenic LM proteins by a novel expression cloning strategy. The antigenic peptides contained within these proteins were defined by deletion analysis of the genes, and their antigenicity was confirmed with synthetic peptides. The nucleotide sequences of the genes showed that they encode previously unknown LM proteins that are homologous to surface proteins in other bacterial species. Consistent with their surface topology, mild trypsin treatment of LM protoplasts ablated T cell recognition of these Ags. These findings establish a general strategy for identifying unknown CD4+ T cell Ags and demonstrate that LM surface proteins can provide the peptides for presentation by MHC class II molecules that are specific targets for CD4+ T cells during murine LM infection. PMID- 9725230 TI - Specific antiviral activity demonstrated by TGTP, a member of a new family of interferon-induced GTPases. AB - The GTPase superfamily includes a diversity of molecules whose functions are regulated through the binding and hydrolysis of GTP. This superfamily can be segregated into families of functionally related molecules that typically share amino acid sequence similarity within and around the nucleotide-binding domains. A new family of putative GTPases, including IRG-47, LRG-47, IGTP, and TGTP/Mg21, has recently emerged that share significant sequence identity (25-40%). Expression of these molecules has been shown to be selectively induced by IFN gamma and in some cases by IFN-alpha beta or bacterial LPS. This induction pattern implicates these putative GTPases as part of the innate defense of cells to infection, but their role in such defense has not yet been defined. We have previously described the cloning of TGTP and now confirm its intrinsic activity as a GTPase. We found that TGTP is strongly induced by endogenous IFN-alpha beta produced in response to standard lipofection of plasmid DNA or polyinosinic polycytidylic acid. The ability of endogenously produced IFN-alpha beta to efficiently induce expression of TGTP under these conditions suggested that TGTP might participate in defense against viral infection. This proposal was borne out when TGTP-transfected L cells displayed relative resistance to plaque formation by vesicular stomatitis virus but not herpes simplex virus. This observation places TGTP among a small family of innate antiviral agents and has implications for the functions of other members of this family of GTPases. PMID- 9725231 TI - Molecular cloning and immunologic reactivity of a novel low molecular mass antigen of Mycobacterium tuberculosis. AB - Polypeptide Ags present in the culture filtrate of Mycobacterium tuberculosis were purified and evaluated for their ability to stimulate PBMC from purified protein derivative (PPD)-positive healthy donors. One such Ag, which elicited strong proliferation and IFN-gamma production, was further characterized. The N terminal amino acid sequence of this polypeptide was determined and used to design oligonucleotides for screening a recombinant M. tuberculosis genomic DNA library. The gene (Mtb 8.4) corresponding to the identified polypeptide was cloned, sequenced, and expressed in Escherichia coli. The predicted m.w. of the recombinant protein without its signal peptide was 8.4 kDa. By Southern analysis, the DNA encoding this mycobacterial protein was found in the M. tuberculosis substrains H37Rv, H37Ra, Erdman, and "C" strain, as well as in certain other mycobacterial species, including Mycobacterium avium and Mycobacterium bovis BCG (bacillus Calmette-Guerin, Pasteur). The Mtb 8.4 gene appears to be absent from the environmental mycobacterial species examined thus far, including Mycobacterium smegmatis, Mycobacterium gordonae, Mycobacterium chelonae, Mycobacterium fortuitum, and Mycobacterium scrofulaceum. Recombinant Mtb 8.4 Ag induced significant proliferation as well as production of IFN-gamma, IL-10, and TNF-alpha, but not IL-5, from human PBMC isolated from PPD-positive healthy donors. Mtb 8.4 did not stimulate PBMC from PPD-negative donors. Furthermore, immunogenicity studies in mice indicate that Mtb 8.4 elicits a Th1 cytokine profile, which is considered important for protective immunity to tuberculosis. Collectively, these results demonstrate that Mtb 8.4 is an immunodominant T cell Ag of M. tuberculosis. PMID- 9725232 TI - Natural killer cell lysis of cytomegalovirus (CMV)-infected cells correlates with virally induced changes in cell surface lymphocyte function-associated antigen-3 (LFA-3) expression and not with the CMV-induced down-regulation of cell surface class I HLA. AB - CMV and other viruses down-regulate the cell surface expression of class I HLA, and while this allows them to evade CTL, it may make infected cells more susceptible to lysis by NK cells, due to the failure to engage class I inhibitory receptors on the NK cell. We studied CMV infection and found that fibroblasts infected with virus strains Towne, Toledo, Davis, and C1FE were refractory to NK lysis, while those infected with strains AD169, C1F, or R7 were susceptible. All viral strains down-regulated class I HLA to a similar extent, and we concluded that there was no evidence for any correlation between the latter and susceptibility to NK lysis. In contrast, there was a strong correlation between NK killing of CMV-infected cells and cell surface levels of lymphocyte function associated antigen-3 (LFA-3). Fibroblasts infected with the Towne, Toledo, Davis, and C1FE strains of CMV down-regulated LFA-3 expression and were refractory to lysis, while strains AD169, C1F, and R7 up-regulated LFA-3 and were susceptible to NK killing. U373 MG (malignant glioma) cells expressed constitutively high levels of LFA-3 and were sensitive to NK lysis when infected with any of the above-listed CMV strains. We estimated that a minimum of between 29,000 and 71,000 LFA-3 molecules per target cell were needed for NK susceptibility. The effects on LFA-3 expression were due to immediate early/early viral gene products. We also demonstrated that fibroblasts infected with the strains Towne, Toledo, Davis, and C1FE expressed a ganciclovir-sensitive late CMV gene product, which delivered an inhibitory signal to NK cells. PMID- 9725233 TI - B and T cells are required for mouse mammary tumor virus spread within the mammary gland. AB - Mouse mammary tumor virus (MMTV) is an infectious retrovirus transmitted through milk from mother to newborns. MMTV encodes a superantigen (SAg) whose activity is indispensable for the virus life cycle, since a genetically engineered virus with a mutation in the sag gene neither amplified in cells of the immune system of suckling pups nor infected their mammary glands. When wild-type MMTV was injected directly into the mammary glands of uninfected pubescent mice, their lymphoid as well as mammary gland cells became virus infected. To test whether this infection of lymphoid cells was dependent on SAg activity and required for virus spread within the mammary gland, we performed mammary gland injections of wild-type MMTV(C3H) into two strains of transgenic mice that lacked SAg-cognate, V beta 14+ T cells. Neither the MTV-ORF or LEL strains showed infection of their mammary glands. Moreover, no MMTV infection of their peripheral lymphocytes was detected. Similar experiments with mice lacking B cells (mu-chain knockouts) showed no detectable virus spread in the mammary glands or lymphoid tissues. These data suggest that SAg activity and MMTV-infected lymphocytes are required, not only for initial steps of viral infection, but also for virus spread within the mammary gland. Virus spread at late times in infection determines whether MMTV induces mammary tumors. PMID- 9725234 TI - Migration inhibitory factor induces killing of Leishmania major by macrophages: dependence on reactive nitrogen intermediates and endogenous TNF-alpha. AB - Macrophage migration inhibitory factor (MIF) is a product of activated T cells, anterior pituitary cells, and macrophages. MIF plays an important role in LPS induced shock and delayed-type hypersensitivity. Furthermore, MIF exhibits a proinflammatory spectrum of action, promoting TNF-alpha production by macrophages, and counter-regulates glucocorticoid suppression of cytokine production. Here, we report that purified recombinant MIF activates murine macrophages to kill Leishmania major, with maximal effects at concentrations above 1 microg/ml. This MIF-mediated activation is specific, since it can be blocked completely by anti-MIF mAb. The MIF-mediated activation is dependent on TNF-alpha produced endogenously by macrophages, because the administration of anti-TNF-alpha antiserum markedly reduced the MIF effect. No MIF-mediated activation was observed in macrophages derived from TNF receptor p55 knockout mice, thus demonstrating the requirement of the smaller TNF receptor molecule for autocrine TNF-alpha signaling. A highly specific inhibitor of the inducible nitric oxide synthase (iNOS), L-N6-(1-iminoethyl)lysine, dihydrochloride, also inhibited the action of MIF, suggesting an important role for iNOS in the antiparasitic properties of MIF. In line with this, no MIF-mediated activation was detected analyzing macrophages derived from iNOS-deficient mice. The effect of MIF was blocked completely by the macrophage-deactivating cytokines IL-10, IL 13, and TGF-beta. Finally, the expression of MIF mRNA and protein was up regulated in lymph nodes of mice during the first week after infection with L. major. MIF therefore represents a cytokine involved not only in the recruitment of proinflammatory cells during infection but also in the complex regulation of the antimicrobial activity of these cells. PMID- 9725235 TI - The mannose receptor mediates induction of IFN-alpha in peripheral blood dendritic cells by enveloped RNA and DNA viruses. AB - Peripheral blood dendritic cells (DC) produce IFN-alpha in response to challenge by many enveloped viruses including herpes simplex virus (HSV) and HIV, whereas Sendai virus predominantly stimulates IFN-alpha production by monocytes. Glycosylated viral envelope proteins are known to be important for the induction of IFN-alpha. In this study we demonstrate that stimulation of IFN-alpha synthesis by HSV is inhibited by a number of monosaccharides, including fucose, N acetylglucosamine, and N-acetylgalactosamine as well as the yeast polysaccharide mannan, supporting a role for lectin(s) in the IFN-alpha stimulation pathway. Furthermore, antiserum to the mannose receptor (MR) also inhibited HSV, vesicular stomatitis virus, and HIV-induced IFN-alpha production, but failed to inhibit the IFN-alpha induced by Sendai virus. We further demonstrated that freshly isolated blood DC and IFN-alpha-producing cells responding to HSV stimulation express the MR. This study therefore implicates the MR as an important receptor for the nonspecific recognition of enveloped viruses by DC and the subsequent stimulation of IFN-alpha production by these viruses. Thus, the MR probably serves as a critical link between innate and adaptive immunity to viruses, especially given the role of the MR in Ag capture by DC and the importance of IFN-alpha in shaping immunity. PMID- 9725236 TI - CTL response to Mycobacterium tuberculosis: identification of an immunogenic epitope in the 19-kDa lipoprotein. AB - The successful resolution of infection with Mycobacterium tuberculosis (M.tb) is believed to involve the induction of CTLs that are capable of killing cells harboring this pathogen, although little information is known about the MHC restriction or fine specificity of such CTLs. In this study, we used knowledge of the HLA-A*0201-binding motif and an immunofluorescence-based peptide-binding assay to screen for potential HLA-A*0201-binding epitopes contained in the 19-kDa lipoprotein of M.tb (M.tb19). CD8+ T cells derived from HLA-A*0201+ patients with active tuberculosis (TB) as well as tuberculin skin test-positive individuals who had no history of TB were used as effector cells to determine whether these epitopes are recognized by in vivo-primed CTLs. An in vitro vaccination system using HLA-A*0201+ dendritic cells (DCs) as APCs was used to determine whether these epitopes can sensitize naive CD8+ T cells in vitro, leading to the generation of Ag-specific CTLs. The results show that an HLA-A*0201-binding peptide comprised of residues 88 to 97 of M.tb19 (P88-97) is recognized by circulating CD8+ CTLs from both healthy tuberculin skin test-positive individuals and patients with active TB but not by tuberculin skin test-negative subjects. Moreover, dendritic cells pulsed with this peptide induced class I MHC-restricted CTLs from the T cells of healthy unsensitized persons. Finally, CTL lines that were specific for P88-97 were shown to lyse autologous monocytes that had been infected acutely with the H37Ra strain of M.tb. These results demonstrate that M.tb19 elicits HLA class I-restricted CTLs in vitro and in vivo that recognize endogenously processed Ag. Epitopes of the type identified here may prove useful in the design of an M.tb vaccine. PMID- 9725237 TI - B7-1, but not CD28, is crucial for the maintenance of the CD4+ T cell responses in human leprosy. AB - We used human leprosy as a model to compare patterns of costimulatory molecule expression in respect to the clinical/immunologic spectrum of disease. We found that B7-1, B7-2, and CD28 transcripts dominated in tuberculoid leprosy patients, who have potent T cell responses to Mycobacterium leprae. In contrast, CTLA-4 was more strongly expressed in lesions from lepromatous patients, who manifest specific T cell anergy to the leprosy bacterium. T cell clones from tuberculoid lesions were CD4+CD28+ or CD4+CD28-, and T cell clones from lepromatous lesions were predominantly CD8+CD28-. The M. leprae-specific recall response of CD4+ T cell clones from tuberculoid lesions was blocked by anti-B7-1 mAb, but not by anti-B7-2 mAb or CTLA-Ig. However, anti-CD28 and anti-CTLA-4 mAbs did not block activation of clones from tuberculoid lesions, suggesting that B7-1 may utilize another costimulatory pathway. Peripheral blood T cell responses in the lepromatous form were strongly regulated by CD28 during T cell activation, in contrast to the tuberculoid form. Thus, B7-1 costimulation could play a role in maintaining a strong immune response to the pathogen. PMID- 9725238 TI - The role of the bacterial membrane protein ActA in immunity and protection against Listeria monocytogenes. AB - ActA, an essential virulence factor of Listeria monocytogenes, is an integral membrane protein that is required for intracellular motility, cell-to-cell spread, and rapid dissemination of the bacteria in the infected host. To reveal cytotoxic T cell responses against ActA we introduced a recombinant soluble form of ActA into the MHC class I-processing compartment of APC using a variant of listeriolysin mutated within its immunodominant MHC class I epitope. With this experimental system we demonstrate that T cells are induced against ActA during a sublethal infection with L. monocytogenes. However, adoptively transferred cytotoxic CD8+ T cells specific for ActA did not protect mice against a subsequent challenge with this pathogen. This was due to an inability of APC to present ActA by either MHC class I or class II molecules as long as ActA remained tethered to the surface of intracellular viable bacteria. ActA was only presented when L. monocytogenes were engineered to secrete ActA or when the bacteria were killed by antibiotics during the assay. These findings raise questions on the general use of membrane proteins of pathogens as candidates for subunit vaccines. PMID- 9725239 TI - Th2-type CD4+ cells neither enhance nor suppress antitumor CTL activity in a mouse tumor model. AB - Many cervical cancers express the E7 protein of human papillomavirus 16 as a tumor-specific Ag (TSA). To establish the role of E7-specific T cell help in CD8+ CTL-mediated tumor regression, C57BL/6J mice were immunized with E7 protein or with a peptide (GF001) comprising a minimal CTL epitope of E7, together with different adjuvants. Immunized mice were challenged with an E7-expressing tumor cell line, EL4.E7. Growth of EL4.E7 was reduced following immunization with E7 and Quil-A (an adjuvant that induced a Th1-type response to E7) or with GF001 and Quil-A. Depletion of CD8+ cells, but not CD4+ cells, from an immunized animal abrogated protection, confirming that E7-specific CTL are necessary and sufficient for TSA-specific protection in this model. Immunization with E7 and Algammulin (an alum-based adjuvant) induced a Th2-like response and provided no tumor protection. To investigate whether a Th2 T helper response to E7 could prevent the development of an E7-specific CTL-mediated protection, mice were simultaneously immunized with E7/Algammulin and GF001/Quil-A or, alternatively, were immunized with GF001/Quil-A 8 wk after immunization with E7/Algammulin. Tumor protection was observed in each case. We conclude that an established Th2 response to a TSA does not prevent the development of TSA-specific tumor protective CTL. PMID- 9725240 TI - CpG DNA induces sustained IL-12 expression in vivo and resistance to Listeria monocytogenes challenge. AB - Vertebrates have evolved innate immune defense mechanisms that recognize and respond to structural patterns that are specific to microbial molecules. One such pattern recognition system is based on unmethylated CpG dinucleotides in particular sequence contexts (CpG motifs); these motifs are common in bacterial DNA but are under-represented ("CpG suppression") and methylated in vertebrate DNA. Mice that are injected with bacterial DNA or synthetic oligodeoxynucleotides (ODNs) containing CpG motifs respond with a rapid production of IL-12 and IFN gamma. The serum levels of IL-12 were increased for at least 8 days after a single injection of CpG ODNs, but IFN-gamma levels returned to baseline within 24 h. This Th1-like cytokine response to CpG motifs induces a state of resistance to infection by Listeria monocytogenes in susceptible specific pathogen-free BALB/c mice. Resistance developed within 48 h of pretreatment with CpG ODNs, persisted for at least 2 wk, and was dependent upon IFN-gamma secretion. These data support the hypothesis that CpG DNA motifs are a "danger signal" that activates protective innate immune defenses and may have therapeutic potential. PMID- 9725241 TI - Lung-specific transgenic expression of KC enhances resistance to Klebsiella pneumoniae in mice. AB - A vigorous host response is required to effectively clear pathogenic bacteria from the lungs and is dependent upon the recruitment and activation of neutrophils and macrophages. A family of chemotactic cytokines, referred to as chemokines, have been shown to participate in this complex protective response. In this study, we assessed the role of the C-X-C chemokine KC in lung antibacterial host defense using wild-type (wt) B6D2 mice or transgenic mice that had been bred on a B6D2 background expressing KC under the control of a Clara cell-specific promoter within the lung. The administration of Klebsiella pneumoniae to both wt and KC-transgenic mice resulted in a time-dependent expression of KC protein within the lung that peaked at 24 to 48 h postinoculation. When infected with K. pneumoniae, the KC-transgenic mice showed a striking improvement in survival compared with wt control mice. This improved survival was due to an increase in bacterial clearance, which occurred in association with a vigorous recruitment of neutrophils in the KC-transgenic mice compared with their wt control counterparts. No differences in the lung levels of the specific cytokines TNF-alpha, IFN-gamma, IL-12, and IL-10 were noted. However, inducible macrophage inflammatory protein-2 levels were significantly decreased in the KC-transgenic mice compared with the wt mice. This study indicates that the compartmentalized overexpression of KC in vivo results in increased lung bacterial clearance and improved survival, which occurs in association with enhanced polymorphonuclear leukocyte influx to the lung. PMID- 9725242 TI - Novel regulation of cyclooxygenase-2 expression and prostaglandin E2 production by IFN-gamma in human macrophages. AB - Cyclooxygenase-2 (COX-2) is the inducible enzyme in macrophages responsible for high output PG production during inflammation and immune responses. Although several stimuli are known to regulate COX-2, the molecular mechanisms modulating its expression by the cytokine network are poorly understood. As IFN-gamma priming is essential for macrophage accessory and effector cell functions, we investigated the effect of IFN-gamma on COX-2 expression in U937 human macrophages stimulated with IL-1beta. A dose- and time-dependent increase in COX 2 mRNA and protein expression was evoked by IL-1beta, whereas the levels of COX 1, the constitutively expressed isoform, remained unaltered. Interestingly, IFN gamma-primed cells showed 40 to 60% lower levels of COX-2 mRNA, protein expression, and PGE2 production as compared with nonprimed cells. IFN-gamma priming (50-500 U/ml) down-regulated COX-2 expression in a time- and dose dependent fashion. Furthermore, IFN-gamma inhibited COX-2 gene transcription in response to IL-1beta but not to LPS. In contrast, the rate of decay of COX-2 transcripts in nonprimed and primed macrophages was similar (t1/2 = 3.2 h). The down-regulatory effect of IFN-gamma on IL-1beta-induced COX-2 expression was abrogated with cycloheximide. These results highlight a novel mechanism of COX-2 regulation by IFN-gamma at the transcriptional level, which may affect the outcome of inflammatory and immune conditions. PMID- 9725243 TI - The roles of L-selectin, beta 7 integrins, and P-selectin in leukocyte rolling and adhesion in high endothelial venules of Peyer's patches. AB - Lymphocyte trafficking into Peyer's patches requires beta 7 integrins and L selectin. Here, we use intravital microscopy to examine leukocyte rolling and adhesion in Peyer's patch high endothelial venules (HEV) of wild-type, L-selectin deficient (L-/-), beta 7 integrin-deficient (beta 7-/-), and beta 7/L(-/-) mice. Although the leukocyte rolling flux fraction was reduced by 70%, Peyer's patches in L-/- mice were of normal size and cellularity. In beta 7-/- mice, the rolling flux fraction was normal, but the number of adherent leukocytes in HEV was greatly reduced. The median leukocyte rolling velocity was reduced in L-/- mice and increased in beta 7-/- mice, suggesting that beta 7 integrins and L-selectin mediate rolling in Peyer's patch HEV at different velocities. beta 7/L(-/-) exhibited both a low rolling flux fraction and low adhesion and had severely reduced Peyer's patch size and cellularity. The residual rolling in these mice was completely blocked by a P-selectin mAb. A significant P-selectin component was also detected in the other genotypes. Twenty-six percent of B and T lymphocytes isolated from Peyer's patches of wild-type mice expressed functional ligands for P-selectin, and this fraction was increased to 57% in beta 7/L(-/-) mice. Peyer's patch HEV were found to express P-selectin under the conditions of intravital microscopy, but not in situ. Our data suggest a novel P-selectin dependent mechanism of lymphocyte homing to Peyer's patches. In situ, beta 7 integrins and L-selectin account for all lymphocyte homing to Peyer's patches, but P-selectin-dependent rolling, as induced by minimal trauma, may support trafficking of effector T lymphocytes to Peyer's patches. PMID- 9725244 TI - IFN-gamma inhibits activation-induced expression of E- and P-selectin on endothelial cells. AB - E- and P-selectin are cell surface lectins that mediate leukocyte-endothelial cell adhesion and thereby participate in neutrophil recruitment into inflammatory sites. E-selectin can be induced on endothelial cells by various activators, including TNF-alpha, IL-1beta, and PMA. Induction of E-selectin is blocked by pretreatment of endothelial cells with IL-4 or TGF-beta, both of which have antiinflammatory properties in vivo. In addition to its well-known proinflammatory activities, IFN-gamma also has antiinflammatory effects in vivo, one of which is inhibition of neutrophil recruitment. To determine whether IFN gamma inhibits neutrophil recruitment by inhibiting adhesion molecule expression, the effect of IFN-gamma on activation-induced cell adhesion molecule expression by cultured HUVEC was evaluated. Pretreatment of endothelial cells with IFN-gamma for 24 to 72 h before 6- to 24-h activation with IL-1beta, TNF-alpha, or PMA resulted in significantly reduced levels of cell surface E-selectin, although levels of ICAM-1 and VCAM-1 were the same or increased. The reduction of cell surface E-selectin levels under these conditions was reflected in reduced levels of E-selectin mRNA, indicating an effect at the transcription level or RNA stability. Interestingly, the increase of cell surface P-selectin expression due to IL-4 treatment of HUVEC was also inhibited by IFN-gamma, while constitutive levels of P-selectin were not. These results suggest that the inhibition of neutrophil recruitment by IFN-gamma in vivo may be due, in part, to the ability of IFN-gamma to inhibit E- and P-selectin up-regulation. Furthermore, these findings emphasize the process of leukocyte recruitment as an important step through which IFN-gamma can direct the character of inflammatory reactions. PMID- 9725245 TI - IL-2 induces T cell adherence to extracellular matrix: inhibition of adherence and migration by IL-2 peptides generated by leukocyte elastase. AB - Migration of inflammatory cells requires cell adhesion and their subsequent detachment from the extracellular matrix (ECM). Leukocyte activation and migration must be terminated to stop inflammation. Here, we report that IL-2 enhances human T cell adherence to laminin, collagen type IV, and fibronectin (FN). In contrast, neutrophil elastase, an enzyme activated during inflammation, degrades IL-2 to yield IL-2 fractions that inhibit IL-2-induced T cell adhesion to FN. The amino acid composition of two of these IL-2 fractions, which appear to block T cell adherence to FN, were analyzed, and three peptides were consequently synthesized. The three peptides IVL, RMLT, and EFLNRWIT, but not the corresponding inversely synthesized peptides, inhibited T cell adhesion to FN induced by a variety of activators: IL-2, IL-7, macrophage inflammatory protein (MIP)-1beta, and PMA, as well as anti-CD3 and anti-beta1 integrin-activating mAb. Moreover, these IL-2 peptides inhibited T cell chemotaxis via FN-coated membranes induced by IL-2 and MIP-1beta. Inhibition of T cell adherence and migration apparently involves abrogation of the rearrangement of the T cell actin cytoskeleton. Thus, the migrating immune cells, the cytokines, and the ECM can create a functional relationship in which both inflammation-inducing signals and inhibitory molecules of immune responses can coexist; the enzymatic products of IL-2 may serve as natural feedback inhibitors of inflammation. PMID- 9725246 TI - Subcellular site of expression and route of vaccination influence pulmonary eosinophilia following respiratory syncytial virus challenge in BALB/c mice sensitized to the attachment G protein. AB - The attachment glycoprotein (G) of respiratory syncytial virus (RSV) is synthesized as two mature forms: a membrane-anchored form and a smaller secreted form. Mutant cDNAs were constructed that encoded one or the other form of the protein and were expressed in recombinant vaccinia viruses (rVV). Mice were immunized with rVV by dermal scarification or i.p. injection to determine the contribution of the membrane-anchored and secreted forms of the G protein on the augmentation of pulmonary pathology seen following RSV challenge. Mice scarified with rVV expressing the membrane-anchored G protein had a markedly reduced pulmonary eosinophilic response following RSV challenge compared with mice scarified with rVV expressing either wild-type or secreted G protein. The induction of pulmonary eosinophilia in rVV-primed mice was also dependent upon the route of vaccination. An eosinophilic response was not observed in any groups of mice immunized i.p. with rVV expressing any of the different forms of the G protein. The difference in pulmonary pathology observed between dermal scarification or i.p. vaccinated mice was not reflected in a difference in cytokine production by splenocytes from vaccinated and challenged mice restimulated with RSV in vitro. Both groups produced significant levels of IL-4 and IL-5. These data suggest that the local APCs and lymphoid environment, together with the form of the G protein, influence pulmonary pathology following RSV challenge. PMID- 9725247 TI - TNF-alpha released by comigrating monocytes promotes transendothelial migration of activated lymphocytes. AB - We investigated mechanisms that increase motility and transendothelial trafficking of activated lymphocytes. Freshly isolated lymphocytes stimulated with immobilized anti-CD3 for 2 h migrate into polymerized collagen in 1.99+/ 0.25-fold greater numbers and across confluent endothelial monolayers in 4.8+/ 0.5-fold greater numbers compared with leukocytes incubated with non-specific IgG. Activated lymphocytes form clusters with monocytes, and their increased motility was dependent on the presence of comigrating monocytes. Five lines of evidence support the idea that monocytes modulate lymphocyte motility through the release of TNF-alpha: 1) flow-cytometric analyses, using highly specific and avid mAbs to probe permeabilized whole blood leukocytes, showed that >80% of circulating monocytes contain intracellular TNF-alpha, whereas <5% contain IL-1 and none contain IL-6; 2) stimulation with immobilized anti-CD3 that was intended to activate lymphocytes also induced monocytes to release increased quantities of TNF-alpha; 3) rTNF-alpha, added in doses of 1 to 20 pg/ml to purified anti-CD3 stimulated lymphocytes, reproduced, in a dose-dependent manner, the motility enhancing effect of adding monocytes; 4) the transient increase in the expression of TNF R-I on CD3-activated T lymphocytes parallels their transiently increased motility; and 5) addition of anti-TNF-alpha, anti-TNF R-I, anti-TNF R-II, or soluble TNF R-I decreased the motility of stimulated lymphocytes. These results suggest that T lymphocyte stimulation via the CD3-TCR complex signals nearby monocytes to release TNF-alpha, which feeds back on the lymphocytes to increase their locomotor activity. PMID- 9725248 TI - Lipopolysaccharide and ceramide use divergent signaling pathways to induce cell death in murine macrophages. AB - Ceramide is a well-known apoptotic agent that has been implicated in LPS signaling. Therefore, we examined whether LPS-induced macrophage cytotoxicity is mediated by mimicking ceramide. Both LPS and the cell-permeable ceramide analogue, C2 ceramide, induced significant cell death in IFN-gamma-activated, thioglycollate-elicited peritoneal macrophages after 48 and 24 h, respectively. Ceramide-induced cell death was neither accompanied by DNA fragmentation nor phosphatidyl serine externalization, characteristics of apoptosis. In contrast, LPS induced a significant fraction of cells to undergo apoptosis, as demonstrated by DNA fragmentation and quantified by DNA analysis on FACS, yet the majority of the cells died in a necrotic fashion. C3H/HeJ Lps(d) macrophages were resistant to LPS-induced cell death and less sensitive to C2 ceramide-evoked cytotoxicity, when compared with Lps(n) macrophages. C2 ceramide plus IFN-gamma failed to activate release of nitric oxide (NO.), whereas LPS-induced cell death, but not C2-induced cytotoxicity, was blocked by an inhibitor of inducible NO. synthase (iNOS), NG-monomethyl-L-arginine. Macrophages from IFN regulatory factor-1 (-/-) mice shown previously to respond marginally to LPS plus IFN-gamma to express iNOS mRNA and NO., were refractory to LPS plus IFN-gamma-induced cytotoxicity and apoptosis. These data suggest that although LPS may mimic certain ceramide effects, signal transduction events that lead to cytotoxicity, as well as the downstream mediators, diverge. PMID- 9725249 TI - Important contributions of P-selectin glycoprotein ligand-1-mediated secondary capture to human monocyte adhesion to P-selectin, E-selectin, and TNF-alpha activated endothelium under flow in vitro. AB - In this study, an in vitro flow model and a blocking mAb to P-selectin glycoprotein ligand-1 (PSGL-1) were used to define the role of PSGL-1 in monocyte attachment and rolling on E- and P-selectin and in attachment and accumulation on 6-h TNF-alpha-activated HUVEC. KPL1, an adhesion-blocking mAb directed against the tyrosine sulfate motif of PSGL-1, abolished monocyte-adhesive interactions with P-selectin, but only partially blocked monocyte interaction with E-selectin. Further analysis showed that on E-selectin, KPL1 blocked only secondary (i.e., monocyte/monocyte) interactions, but did not block primary (i.e., monocyte/E selectin) interactions, with secondary adhesion accounting for 90% of the total adhesive interactions on either E- or P-selectin. On cytokine-activated HUVEC, monocytes initially attached and formed linear strings of adherent cells, which involved both primary and secondary adhesion. PSGL-1 or L-selectin mAb reduced string formation, and the combination of PSGL-1 and L-selectin mAb prevented monocyte strings and inhibited 86% of accumulation. Monocyte attachment and rolling on purified adherent monocytes were also critically dependent on PSGL-1 on the adherent monocytes. These studies document that secondary interactions between monocytes, mediated by PSGL-1, are crucial for monocyte initial attachment, rolling, and accumulation on activated endothelium under laminar shear flow. PMID- 9725250 TI - Bradykinin stimulates IL-8 production in cultured human airway smooth muscle cells: role of cyclooxygenase products. AB - IL-8 is an important neutrophil and eosinophil chemoattractant in asthma. A recent report has suggested that bradykinin (BK), an asthmatic mediator, induces the release of IL-8 in nonairway cells. We have recently reported that BK causes cyclooxygenase (COX)-2 induction and PGE2 release in human airway smooth muscle (ASM) cells. In this study, we tested the ability of BK to induce IL-8 from these cells and explored the role of COX products and COX-2 induction in this process. Confluent serum-deprived human ASM cells were studied. IL-8 was assayed by specific ELISA. Unstimulated cells released low levels of IL-8. BK enhanced IL-8 release in a concentration- and time-dependent fashion (maximum 50-fold increase over basal). The nonselective COX inhibitor indomethacin and the selective COX-2 inhibitor NS-398 strongly inhibited BK-stimulated PGE2 and IL-8 production. The COX substrate arachidonic acid also caused PGE2 and IL-8 production, and its effect was inhibited by nonselective COX inhibitors but unaffected by NS-398. Both the BK- and arachidonic acid-induced IL-8 production was inhibited by the protein synthesis inhibitors cycloheximide and actinomycin D and by the steroid dexamethasone. Furthermore, exogenous PGE2 and calcium ionophore A23187 also stimulated IL-8 release. BK-induced IL-8 release was mimicked by the BK B2 receptor agonist (Tyr(Me)8)-BK and was potently inhibited by the selective B2 receptor antagonist HOE-140. These results suggest that human ASM can be a source of IL-8 and also that endogenous prostanoids, involving both COX-1 and COX-2, have a novel role in mediating BK-induced IL-8 production. PMID- 9725251 TI - Contribution of endothelial selectins and alpha 4 integrins to eosinophil trafficking in allergic and nonallergic inflammatory reactions in skin. AB - The role of endothelial selectins in mediating eosinophil recruitment was assessed using the trafficking of 111In-labeled blood eosinophils in mouse skin. An intradermal injection of chemoattractants (leukotriene B4, macrophage inflammatory protein-l alpha, and eotaxin) resulted in a rapid accumulation of 111In eosinophils that was reduced 49 to 91% by anti-P-selectin mAb. An anti-E selectin mAb was ineffective, although a combined E- and P-selectin blockade resulted in >95% inhibition of all responses. The accumulation of a pulse of 111In eosinophils at sites of active cutaneous anaphylaxis (ACA) at 4 to 8 h and at 20 to 24 h after Ag challenge was completely dependent upon E- and P-selectin in combination, but not in isolation. In contrast, at 20 to 24 h after Ag challenge in a delayed-type hypersensitivity (DTH) reaction in skin, 111In eosinophil accumulation was largely independent of endothelial selectins, even when L-selectin was also blocked. An anti-alpha 4 integrin mAb significantly reduced 111In eosinophil trafficking in both allergic reactions but was slightly more effective in the DTH reaction compared with the ACA reaction. These results show that P-selectin and to a lesser extent E-selectin mediate eosinophil recruitment in skin in acute inflammatory reactions. In allergic, late-onset inflammatory reactions, neither P- nor E-selectin alone are sufficient to mediate eosinophil accumulation; when combined, they are essential for trafficking in ACA but are less important in the DTH reaction. Whether alpha 4 integrin-based strategies will be more effective than selectin-based strategies at inhibiting eosinophil recruitment in human disease remains to be determined. PMID- 9725252 TI - Advanced oxidation protein products as novel mediators of inflammation and monocyte activation in chronic renal failure. AB - We previously demonstrated the presence of advanced oxidation protein products (AOPP), a novel marker of oxidative stress in the plasma of uremic patients receiving maintenance dialysis. The present study in a cohort of 162 uremic patients showed that plasma concentrations of AOPP increased with progression of chronic renal failure and were closely related to advanced glycation end products (AGE)-pentosidine (r = 0.52, p < 0.001), taken as a marker of AGE. In vivo, the relevance of AOPP and AGE-pentosidine in monocyte-mediated inflammatory syndrome associated with uremia was evidenced by close correlations between AOPP or AGE pentosidine and monocyte activation markers, including neopterin, IL-1R antagonist, TNF-alpha, and TNF soluble receptors (TNF-sR55 and TNF-sR75). To determine the mechanisms by which AOPP and AGE could be directly involved in monocyte activation, AOPP-human serum albumin (HSA) and AGE-HSA were produced in vitro by treating HSA with oxidants or glucose, respectively. Spectroscopic analysis confirmed that AOPP-HSA contains carbonyls and dityrosine. Both AOPP-HSA and AGE-HSA, but not purified dityrosine, were capable of triggering the oxidative burst of human monocytes in cultures. The AOPP-HSA-induced respiratory burst was dependent on the chlorinated nature of the oxidant and on the molar ratio HSA/HOCI. Collectively, these data first demonstrate that AOPP act as a mediator of oxidative stress and monocyte respiratory burst, which points to monocytes as both target and actor in the immune dysregulation associated with chronic uremia. PMID- 9725253 TI - Effect of human C-reactive protein on chemokine and chemotactic factor-induced neutrophil chemotaxis and signaling. AB - C-reactive protein (CRP) is a unique serum pentraxin and the prototype acute phase reactant. CRP is a ligand for specific receptors on phagocytic leukocytes, and mediates activation reactions of monocytes/macrophages, but inhibits the respiratory burst of neutrophils (PMN). Since CRP selectively accumulates at inflammatory sites in which IL-8 is also produced, we tested the effects of CRP on the responsiveness of PMN to IL-8 and the bacterial chemotactic peptide, FMLP phenylalanine (FMLPP). Purified human CRP inhibited the chemotactic response of PMN to IL-8 and FMLPP. A mouse IgM mAb that was generated against the leukocyte CRP receptor (CRP-R) also inhibited the chemotactic response. Incubation of purified CRP with activated PMN generated CRP-derived peptides that also inhibited chemotaxis. A synthetic CRP peptide (residues 27-38) that binds to the CRP-R had weak chemotactic activity, whereas two other CRP synthetic peptides (residues 174-185 and 191-205) inhibited chemotaxis of PMNs to both IL-8 and FMLPP. CRP did not alter receptor-specific binding of IL-8, but exerted its effect at the level of signaling. CRP augmented both IL-8- and FMLPP-induced mitogen-activated protein kinase (extracellular signal-regulated kinase-2) activity. CRP at acute phase levels increased both agonist-induced and noninduced phosphatidylinositol-3 kinase activity. The results suggest a role for CRP as a regulator of leukocyte infiltration at inflammatory sites. PMID- 9725254 TI - Short-term and long-term cytokine release by mouse bone marrow mast cells and the differentiated KU-812 cell line are inhibited by brefeldin A. AB - Mast cells and basophils produce a wide range of cytokines, including large amounts of both IL-6 and granulocyte-macrophage CSF (GM-CSF). However, the route by which cytokines are secreted is poorly understood. In the current study, we used two inhibitors of vesicular transport, brefeldin A and monensin, to examine the routes of secretion of IL-6 and GM-CSF in the differentiated KU812 human cell line and cultured mouse bone marrow mast cells (mBMMC). Studies of cytokine production over 6 to 24 h demonstrated that IL-6 and GM-CSF release from both cell types were inhibited by brefeldin A (BFA) following activation with calcium ionophore, A23187. Monensin had similar inhibitory effects to that of BFA on the initial and ongoing IL-6 release from KU812 cells. In contrast, the amount of each cytokine remaining within the cells was significantly enhanced. Similar results were obtained following IgE-mediated activation of mBMMC. BFA significantly inhibited both the constitutive secretion of IL-6 and the immediate ionophore-induced increase in IL-6 release from KU812 cells at 20 min postactivation. However, treatment with these agents did not alter the release of histamine and beta-hexaminidase from either mBMMC or KU812 cells. These studies suggest that both the initial 20-min release of IL-6 and secretion of IL-6 and GM CSF over up to 24 h by mBMMC and differentiated KU-812 cells occur predominately through a vesicular transport-dependent mechanism, and that little, if any, IL-6 and GM-CSF is released through degranulation. PMID- 9725255 TI - Differential regulation of lipopolysaccharide (LPS) activation pathways in mouse macrophages by LPS-binding proteins. AB - LPS binding to its receptor(s) on macrophages induces the synthesis of inflammatory mediators involved in septic shock. While the signaling mechanism(s) remains to be fully defined, the human LPS-binding protein (LBP) is known to regulate responses to LPS by facilitating its binding to CD14 on human monocytes. The structurally related bactericidal permeability increasing protein (BPI) differs from LBP by inhibiting LPS-induced human monocyte activation. We have demonstrated that, unlike the human monocyte response to LPS, both LBP and BPI inhibited LPS-stimulated TNF-alpha production in mouse peritoneal macrophages. In contrast, LPS-dependent nitric oxide release was not affected by LBP. LPS induces the phosphorylation of a number of proteins in a dose and time-dependent manner, however, the pattern of LPS-induced phosphorylation was not reduced by either LBP or BPI under conditions that result in selective TNF-alpha inhibition. Further, activation of the transcription factor NF-kappaB in response to LPS was also not modified by either LBP or BPI. Finally, no differences were detected in TNF-alpha or inducible nitric oxide synthase mRNA accumulations induced by LPS in the presence or absence of either protein, whereas a slight decreased mRNA stability was observed in the group with LPS treatment. These results would suggest that many of the early signaling events contribute to LPS-induced macrophage signaling at a point preceding the divergence of pathways that differentially regulate TNF alpha and NO production. PMID- 9725256 TI - The influence of mast cells on pathways of transepithelial antigen transport in rat intestine. AB - Luminal Ag challenge of intestinal segments from sensitized rats results in a rapid (approximately 3 min) secretory response. We previously showed in horseradish peroxidase (HRP)-sensitized rats that the initial phase of transepithelial Ag transport occurred via a transcellular route and was enhanced by sensitization. However, following the hypersensitivity reaction, Ag also crossed between epithelial cells. The aim of this study was to determine the role of mast cells in the altered transepithelial Ag transport. White spotting mast cell-deficient rats and +/+ littermate controls were sensitized to HRP. After 10 to 14 days, jejunal segments were resected, mounted in Ussing chambers, and challenged with HRP on the luminal side. Electron microscopy of jejunum fixed at 2 min showed a similarly enhanced endocytic transport of HRP in sensitized +/+ and Ws/Ws rats compared with naive controls. In sensitized +/+ rats, a secretory response occurred approximately 3 min after challenge, and tissue conductance increased thereafter. Naive +/+ and sensitized Ws/Ws rats did not demonstrate a secretory response to HRP challenge, and conductance remained at baseline levels. The flux of HRP was elevated across tissue from sensitized +/+ rats but not across tissue from naive controls or sensitized Ws/Ws rats. The results indicate that sensitization enhances the initial phase of transepithelial uptake of Ag by transcytosis in a mast cell-independent manner. However, subsequent recruitment of the paracellular pathway for Ag transport in sensitized rats is dependent upon the presence of mast cells and occurs after the activation of such cells. PMID- 9725257 TI - The up-regulation of IL-6 and IL-8 in human endothelial cells by activated protein C. AB - The protein C/protein S anticoagulant pathway has been proposed to be a common link between coagulation and inflammation. Studies have suggested that a component of the anticoagulant pathway, activated protein C (APC), may play a role in the inflammatory response by modulating the effects of cytokines such as TNF and by blocking neutrophil activation. Cytokines are known to be intimately involved in the inflammatory response and to function in part to restore hemostatic balance. To begin to delineate what role APC may have in the inflammatory response, we have investigated the effect of APC on the production of the proinflammatory cytokines IL-6 and IL-8 in primary HUVEC, human microvascular endothelial cells, and human coronary artery endothelial cells. Our results have demonstrated that physiologic concentrations of APC significantly up regulated the production of both IL-6 and IL-8. This increase, which was seen at both the RNA and protein level, was not due to either thrombin or LPS contamination of the APC preparation. Additional studies also showed that the APC mediated up-regulation of IL-6 and IL-8 was IL-1 independent. Although neither purified protein C nor protein S alone had an effect on cytokine production, protein S, the cofactor for APC, significantly enhanced the ability of APC to up regulate IL-6/IL-8 production. These results provide further evidence for a role for APC in the inflammatory response. PMID- 9725258 TI - Intracellular expression of Fc gamma RIII (CD16) and its mobilization by chemoattractants in human eosinophils. AB - We characterized the existence, translocation, and reabsorption during cellular activation of a constitutively expressed intracellular CD16 in the human eosinophil. By two-color flow cytometry, we showed that 6.5+/-0.3% of nonpurified eosinophils expressed surface CD16. After digestion with phosphatidylinositol specific phospholipase C, surface CD16 on both neutrophils and eosinophils decreased substantially, suggesting that eosinophil CD16 is a glycosyl phosphatidylinositol-linked isoform. However, CD16 was substantially expressed intracellularly in human eosinophils. Epitope-specific binding to CLB-gran11 mAb from non-NA2/NA2 donors demonstrated that intracellular eosinophil CD16 also differed from the transmembrane isoform of CD16 expressed on NK cells or macrophages. Western blot analysis performed with 3G8 or DJ130c mAb showed a broad band at approximately 65 to 80 kDa, which was the same as neutrophil CD16 from the same NA2/NA2 donors. Upon stimulation by chemoattractants C5a, FMLP, or platelet-activating-factor, eosinophilic intracellular CD16 was rapidly translocated to the eosinophil surface, expressed maximally at 30 s, and then gradually disappeared from the cell surface during the next 10 min. Intracellular flow cytometry of stimulated eosinophils and sandwich ELISA of stimulated eosinophil supernatants demonstrated that the disappearance was due to its rapid release into medium and reabsorption by the cells. Our data identify a CD16B that is consistently expressed intracellularly but only rarely on the surface of nonactivated human eosinophils. This CD16 is transiently expressed during stimulation by chemoattractants. PMID- 9725259 TI - Macrophage-inflammatory protein-3 beta/EBI1-ligand chemokine/CK beta-11, a CC chemokine, is a chemoattractant with a specificity for macrophage progenitors among myeloid progenitor cells. AB - Chemoattractants are potential factors influencing cell migration. Stromal cell derived factor-1, a CXC chemokine, is the only chemokine reported to have chemotactic activity for hemopoietic progenitor cells (HPC). We report in this work another chemokine of the CC subfamily, which is chemotactic for HPC. Macrophage-inflammatory protein (MIP)-3 beta/EBI1-ligand chemokine/CK beta-11 attracted bone marrow and cord blood CD34+ cells. In contrast to stromal cell derived factor-1, which attracts multiple types of HPC, MIP-3beta attracted mainly CFU granulocyte macrophage, but not other HPC such as burst-forming unit erythrocyte or CFU granulocyte, erythrocyte, macrophage, and megakaryocyte. Chemoattracted CD34+ cells formed CFU granulocyte macrophage-like colonies, which were morphologically determined as large macrophages. These progenitors were selectively responsive to stimulation by macrophage CSF, demonstrating that MIP-3 beta attracts macrophage progenitors. Expression of CCR7, the receptor for MIP-3 beta, was detected at a mRNA level in the attracted CD34+ cells as well as input CD34+HPC. Expression of MIP-3 beta mRNA was not constitutive, but was inducible in bone marrow stromal cells by inflammatory agents such as bacterial LPS, IFN gamma, and TNF-alpha. Taken together, our findings suggest that MIP-3 beta is expressed in the bone marrow environment after induction with certain inflammatory cytokines and LPS, and may play a role in trafficking of macrophage progenitors in and out of the bone marrow in inflammatory conditions. PMID- 9725260 TI - Histamine potently suppresses human IL-12 and stimulates IL-10 production via H2 receptors. AB - IL-12 and IL-10, respectively, stimulate Th1 and Th2 immune responses. The development of some allergic reactions, infections, and tumors are associated with excessive histamine production and a shift toward Th2 responses. Here we address the possibility that this association is causally linked, at least in part, to modulation of IL-12 and IL-10 production by histamine. We report that histamine dose-dependently inhibited the secretion of human IL-12 (p70) and increased the production of IL-10 in LPS-stimulated whole blood cultures. These effects of histamine were antagonized by cimetidine, an H2 receptor antagonist, but not by selective H1 and H3 receptor blockers, and were mimicked by an H2 receptor agonist. The effects of histamine on IL-12 and IL-10 secretion were independent of endogenous secretion of IL-10 or exogenous addition of IL-12, while Ro 20-1724, a phosphodiesterase inhibitor, potentiated the effects of histamine on IL-12 and IL-10 production, implicating cAMP in its actions. Similar modulatory effects of histamine on IL-12 and IL-10 production, which were reversed by the H2 antagonist cimetidine, were observed in PBMC and isolated monocytes stimulated by Staphylococcus aureus Cowan strain 1 and LPS, respectively. Thus, histamine, via stimulation of H2 receptors on peripheral monocytes and subsequent elevation of cAMP, suppresses IL-12 and stimulates IL-10 secretion, changes that may result in a shift of Th1/Th2 balance toward Th2 dominance. This may represent a novel mechanism by which excessive secretion of histamine potentiates Th2-mediated allergic reactions and contributes to the development of certain infections and tumors normally eliminated by Th1-dependent immune mechanisms. PMID- 9725261 TI - The heat shock response inhibits RANTES gene expression in cultured human lung epithelium. AB - The chemokine RANTES is thought to be involved in the pathophysiology of inflammation-associated acute lung injury. Although much is known regarding signals that induce RANTES gene expression, relatively few data exist regarding signals that inhibit RANTES gene expression. The heat shock response, a highly conserved cellular defense mechanism, has been demonstrated to inhibit a variety of lung proinflammatory responses. We tested the hypothesis that induction of the heat shock response inhibits RANTES gene expression. Treatment of A549 cells with TNF-alpha induced RANTES gene expression in a concentration-dependent manner. Induction of the heat shock response inhibited subsequent TNF-alpha-mediated RANTES mRNA expression and secretion of immunoreactive RANTES. Transient transfection assays involving a RANTES promoter-luciferase reporter plasmid demonstrated that the heat shock response inhibited TNF-alpha-mediated activation of the RANTES promoter. Inhibition of NF-kappaB nuclear translocation with isohelenin inhibited TNF-alpha-mediated RANTES mRNA expression, indicating that RANTES gene expression is NF-kappaB dependent in A549 cells. Induction of the heat shock response inhibited degradation of the NF-kappaB inhibitory protein, I kappaBalpha but did not significantly inhibit phosphorylation of I-kappaBalpha. We conclude that the heat shock response inhibits RANTES gene expression by a mechanism involving inhibition of NF-kappaB nuclear translocation and subsequent inhibition of RANTES promoter activation. The mechanism by which the heat shock response inhibits NF-kappaB nuclear translocation involves stabilization of I kappaBalpha, without significantly affecting phosphorylation of I-kappaBalpha. PMID- 9725262 TI - Expression and functional activity of the IL-8 receptor type CXCR1 and CXCR2 on human mast cells. AB - To further elucidate mechanisms involved in mast cell accumulation at sites of cutaneous inflammation, we have studied the ability of human leukemic mast cells (HMC-1 cells) to express functionally active IL-8 receptors. Expression of mRNA for both types of IL-8 receptors (CXCR1 and CXCR2) was demonstrated by PCR and of both proteins by flow cytometry. Binding and competition studies with 125I labeled IL-8 and its homologue melanoma growth stimulating activity (125I-labeled MGSA) revealed two specific binding sites for IL-8, K1 = 1.1 x 10(11) M(-1) and K2 = 5 x 10(7) M(-1); and for MGSA, K1 = 2.8 x 10(10) M(-1) and K2 = 5 x 10(7) M( 1). This finding was supported by a dose-dependent rise of cytosolic free calcium concentration ([Ca2+]i) induced by both chemokines and to a lesser extent by the homologue neutrophil-activating peptide-2 (NAP-2). A significant migratory response of human leukemic mast cells (HMC-1) was observed with all three chemokines at a range from 10(-8) M to 10(-9) M. Moreover, the formation of cellular F-actin was induced in a rapid, dose-dependent fashion, with a maximally 1.7-fold increase at 10(-7) M. Using postembedding immunoelectron microscopy, we could show the expression of CXCRI on the cytoplasmatic membrane of isolated human skin mast cells whereas CXCR2 was located in mast cell-specific granules. These findings demonstrate for the first time the functional expression of both types of IL-8 receptors on human mast cells, suggesting a role for their ligands during mast cell activation and recruitment. PMID- 9725263 TI - Ex vivo anti-CD3 antibody-activated donor T cells have a reduced ability to cause lethal murine graft-versus-host disease but retain their ability to facilitate alloengraftment. AB - The purpose of this study was to determine whether ex vivo anti-CD3 Ab-activated T cells behaved in a biologically similar manner as naive T cells with respect to causing graft-vs-host disease (GVHD) and facilitating engraftment after allogeneic marrow transplantation. This question was addressed using two well defined MHC-incompatible murine models of GVHD (C57BL/6 (H-2b)-->BIO.BR (H-2k)) and engraftment (C57BL/6 (H-2b)-->AKR/J (H-2k)). Transplantation with anti-CD3 activated T cells significantly reduced GVHD compared with that in animals transplanted with equivalent numbers of naive T cells. Protection from GVHD was not T cell subset dependent, as highly enriched populations of either activated CD4+ or CD8+ T cells caused less lethal GVHD than comparable numbers of purified naive CD4+ or CD8+ T cells. Transplantation with activated T cells also resulted in protection from LPS-mediated GVH lethality in unirradiated F1 recipients. Analysis of immune recovery indicated that animals transplanted with activated T cells had thymic and splenic B cell reconstitution that compared favorably to that in non-GVHD control mice. When engraftment was analyzed, equivalent degrees of donor cell engraftment were observed when animals were transplanted with limiting numbers (5 x 10(5)) of naive vs activated B6 T cells. Further studies indicated that activated CD8+ T cells were exclusively responsible for enhancing engraftment and that facilitation of engraftment was dependent upon the direct recognition of host MHC alloantigens. Collectively, these data demonstrate that transplantation with anti-CD3 Ab-activated T cells results in a reduction in GVHD, but these cells retain their ability to facilitate alloengraftment. The use of this approach in allogeneic marrow transplantation may represent an alternative strategy to mitigate GVHD without compromising engraftment. PMID- 9725264 TI - Mature mainstream TCR alpha beta+CD4+ thymocytes expressing L-selectin mediate "active tolerance" in the nonobese diabetic mouse. AB - Pathogenic autoreactive T lymphocytes are mediators of spontaneous insulin dependent diabetes in nonobese diabetic (NOD) mice. This is demonstrated by their capacity to transfer diabetes into syngeneic immunoincompetent recipients. In addition, especially in prediabetic NOD mice, peripheral CD4+ T lymphocytes were identified that are highly effective, in conventional mixing cotransfer experiments, at preventing disease transfer. The present data demonstrate that mature heat-stable Ag-TCR alpha beta+CD8-thymocytes from prediabetic NOD mice also express this inhibitory capacity. Selection using an L-selectin (CD62L) specific Ab showed that TCR alpha/beta+CD4+CD62L+ thymocytes, emerging from the mainstream differentiation pathway, concentrate this ability to regulate autoreactive effectors. Compared with mature TCR alpha beta+CD8- thymocytes, significantly lower numbers of TCR alpha beta+CD4+CD62L+ were sufficient to achieve an efficient inhibition of disease transfer into NOD-scid recipients. This protective ability was potentiated following in vitro culture in the presence of IL-7. In contrast, TCR alpha beta+CD62L- thymocytes, highly enriched in class I-restricted NK T cells, were unable to influence diabetes transfer. Identical results were obtained using thymocytes that have been cultured in vitro for 4 days in the presence of IL-7. These results support the active role in NOD mice of a thymus-derived CD4+ subset that controls peripheral pathogenic autoimmune effectors. PMID- 9725265 TI - Impaired yield, phenotype, and function of monocyte-derived dendritic cells in humans at risk for insulin-dependent diabetes. AB - Dendritic cells (DC) present Ag to naive T cells and are therefore pivotal in shaping immune responses. DC may either immunize or tolerize T cells. Humans with pancreatic islet autoimmunity at high risk for insulin-dependent diabetes mellitus (IDDM) present the opportunity to investigate DC in autoimmune disease. We compared DC phenotype and function in 12 euglycemic, asymptomatic IDDM relatives with islet autoimmunity and controls matched for age, sex, and MHC class II alleles. DC were generated from adherent peripheral blood cells by culture with granulocyte/macrophage-CSF and IL-4. The yield of DC was significantly lower in IDDM relatives than in controls. While the DC phenotype, HLA-DR+CD14-, was expressed by > or =90% of the cells generated from relatives and controls, the proportion of cells that expressed CD1a and the costimulator molecules CD80 (B7-1) and CD86 (B7-2) was significantly lower in IDDM relatives. In addition, B7-1 and B7-2 expression per cell was significantly lower in IDDM relatives. These phenotypic changes were accompanied by reduced stimulation of autologous CD4 cells by DC from IDDM relatives. Similar findings were obtained in three recently diagnosed IDDM patients. These findings indicate that impairment of DC phenotype and function is a marker of islet autoimmunity and are consistent with a role for impaired DC function in the pathogenesis of autoimmune disease. PMID- 9725266 TI - Pathogenic Mycobacterium tuberculosis evades apoptosis of host macrophages by release of TNF-R2, resulting in inactivation of TNF-alpha. AB - Infection by Mycobacterium tuberculosis (MTB) induces human alveolar macrophage (AMphi) apoptosis by a TNF-alpha-dependent mechanism. The apoptotic response is postulated to be a defense mechanism, limiting the growth of this intracellular pathogen. Consistent with that model, recent studies showed that the virulent MTB strain H37Rv induces substantially less AMphi apoptosis than the attenuated strain H37Ra. We now report that AMphi infection with either H37Rv or H37Ra induces comparable levels of TNF-alpha measured by ELISA but that TNF-alpha bioactivity is reduced in supernatants of H37Rv-infected AMphi. Differential release of soluble TNFR2 (sTNFR2), with formation of inactive TNF-alpha-TNFR2 complexes accounted for the difference in TNF-alpha bioactivity in these cultures. Release of sTNFR2 by H37Rv-infected AMphi was IL-10 dependent since it was inhibited by neutralizing anti-IL-10 Ab. Thus, the effect of TNF-alpha produced by AMphi following infection can be modulated by virulent MTB, using IL 10 as an upstream mediator. PMID- 9725267 TI - cDNA encoding a single-chain antibody to HIV p17 with cytoplasmic or nuclear retention signals inhibits HIV-1 replication. AB - HIV-1 gag p17 protein is an attractive target for molecular intervention, because it is involved in the viral replication cycle at both the pre- and postintegration levels. In the present experiments, we targeted p17 by intracellularly expressing a cDNA encoding an Ab to p17. cDNA from a hybridoma secreting Ab to p17 was cloned, sequenced, reconstructed as a single-chain Ab fragment (scFv), and expressed in the cytoplasm or nucleus with appropriate retention signals. The expressed scFvs had no effect on T cell growth or CD4 expression and bound specifically to HIV-1 p17. Human CD4+ Jurkat T cells that expressed scFvs and were infected with HIV-1 showed a marked reduction in virus replication compared with cells expressing vector alone. The inhibition of virus replication was more pronounced when scFvs were expressed in the cytoplasm rather than the nucleus. From these studies, we conclude that the intracellular expression of a single-chain Ab to p17 inhibits HIV replication; in addition, the degree of inhibition is related to the intracellular targeting site. PMID- 9725269 TI - Increased susceptibility to postoperative sepsis in patients with impaired monocyte IL-12 production. AB - IL-12 is a potent immunoregulatory cytokine that is essential for the development of protective immunity, as demonstrated by numerous animal models of infection. Here, we provide evidence for a critical role of IL-12 in human sepsis. The results of a prospective study of 184 patients undergoing major elective surgery of the upper and lower gastrointestinal tract revealed that, in contrast to patients showing uneventful recovery, monocyte IL-12 production was severely and selectively impaired in patients developing postoperative sepsis. Moreover, the extent of monocyte IL-12 suppression correlated with the severity of postoperative sepsis. Monocyte IL-12 secretion was suppressed before surgery and remained low until the onset of sepsis. Therefore, the suppression of IL-12 secretion preceded the onset of postoperative sepsis but did not occur as a consequence of major surgery. In contrast, IL-1beta production was only reduced during the late postoperative course in patients developing postoperative sepsis, and TNF-alpha release was even increased at different time intervals before the onset of sepsis. Thus, reduced IL-12 release does not reflect a general defect in monocyte cytokine production. Consequently, these results establish a critical role for IL-12 in early resistance to postoperative infection and may allow for the development of novel therapeutic strategies designed to stimulate host defense mechanisms and to reduce the incidence and severity of septic complications. PMID- 9725268 TI - IA-2 (islet cell antigen 512) is the primary target of humoral autoimmunity against type 1 diabetes-associated tyrosine phosphatase autoantigens. AB - IA-2 (islet cell Ag 512) and IA-2 beta (phogrin/IAR) are related autoantigens associated with type 1 diabetes. To determine the critical regions for autoantibody binding and which of these autoantigens is the primary target, mutant and chimeric constructs were used to characterize Ab epitope binding in sera from 217 new onset patients with type 1 diabetes and sequential samples from 141 islet cell Ab positive first degree relatives of patients. All 22 relatives and 121 of 129 patients with IA-2/IA-2 beta Abs had reactivity to IA-2-specific epitopes. These epitopes were in the juxtamembrane region (residues 601-682) and the protein tyrosine phosphatase (PTP)-like domain of IA-2. Chimeras showed that IA-2 residues 795-889 were important for IA-2-specific Ab binding in the PTP-like domain, and mutation of IA-2 residues 877 and 911, previously indicated as relevant for phosphatase activity, also reduced Ab binding. In contrast, Ab binding to IA-2 beta was limited to its PTP-like domain, most IA-2 beta Abs recognized epitopes shared with IA-2, and only 20 patients and 2 relatives had Abs to IA-2 beta-specific epitopes. In 4 relatives, IA-2 and/or IA-2 beta Abs developed in follow-up samples. In each of these, Abs to IA-2-specific epitopes were the first detected. In three, spreading to epitopes shared between IA-2 and IA-2 beta in subsequent samples was seen. In the 17 relatives who developed type 1 diabetes, progression to disease was associated with reactivity to multiple IA 2/IA-2 beta epitopes. These data suggest that IA-2 is the primary phosphatase like autoantigen associated with type 1 diabetes and that studying autoantibody epitope diversity may assist in disease prediction. PMID- 9725271 TI - Eugene W. Caldwell lecture. Dream no small dreams. PMID- 9725270 TI - ARRS presidential address. Health care challenge. American Roentgen Ray Society. PMID- 9725272 TI - 1998 ARRS President's Award. The potential of in vivo vascular tissue engineering for the treatment of vascular thrombosis: a preliminary report. American Roentgen Ray Society. AB - OBJECTIVE: Current gene therapy and tissue engineering protocols suffer from a number of inherent limitations. In this study, we examine the feasibility of a new approach for the treatment of vascular thrombosis: in vivo tissue engineering. MATERIALS AND METHODS: Rabbit femoral veins were transfected in situ with either a previously characterized adenoviral-construct-expressing tissue plasminogen activator or a viral (adenoviral-construct-expressing beta galactosidase) or nonviral (buffer) control and used as cross sections (n = 3). Treated veins were then harvested and grafted into the ipsilateral common femoral artery as an interposition vein graft. A potent stimulus for thrombus formation was then introduced into the recipient artery downstream of the graft. Six days later, the rabbits were sacrificed, and the grafts and downstream arteries were harvested. Vessel segments were then examined for thrombus according to defined anatomic zones. Transfection efficiency and presence of smooth muscle cells in the vein graft were also evaluated. RESULTS: The engineered vein graft showed a significant reduction in thrombus formation within both the graft and the downstream artery relative to nonviral (buffer) and viral (adenoviral-Rous sarcoma virus beta-galactosidase [Adv/RSV-betagal]) controls. Underlying endothelial cell transfection efficiency of 90% was observed in viral controls (Adv/RSV-betagal). A 2.4-fold increase in smooth muscle alpha-actin positive cells in the engineered vein graft was seen compared with nonviral (phosphate buffered saline) controls. A 10-fold increase in smooth muscle alpha-actin positive cells in the engineered vein graft relative to viral (Adv/RSV-betagal) controls was also observed. CONCLUSION: In vivo tissue engineering is a new paradigm in molecular medicine that is a viable alternative to conventional gene therapy and tissue engineering for the treatment of vascular thrombosis. PMID- 9725273 TI - 1998 ARRS Executive Council Award. Radiology in the emergency department: technique for quantitative description of use and results. American Roentgen Ray Society. AB - OBJECTIVE: We sought to develop quantitative methods to describe the use and results of imaging studies in emergency department patients. MATERIALS AND METHODS: A computerized nonrelational database containing records of 3.5 million diagnostic reports generated by our radiology department from 1988 to 1997 was queried using Boolean and natural language search tools. Each record contained data fields for patient demographics, examination description and billing code, names of interpreting radiologists and referring physicians, patient history, report body, and report impression. RESULTS: Emergency department admissions and imaging studies were stable from 1991 to 1997, averaging 60,000 and 52,000 per year, respectively. Bone radiographs comprised 45.1% of examinations; chest radiographs, 44.6%; and abdominal radiographs, 10.4%. The percentages of radiographs interpreted as normal were 75.9% in 1992 and 75.3% in 1996, with cervical spine (88.7%), thoracic spine (86.3%), and knee (86.3%) yielding the highest proportion of studies with normal findings. The number of CT studies of the body increased from 1840 in 1993 to 3101 in 1997. Studies of the abdomen accounted for most of this increase (52.3% in 1993 to 66.0% in 1997). During evaluations for cervical spine injury, a mean of 6.5% of radiographic studies were followed by CT studies, and the findings of 89.0% of those CT studies were interpreted as normal. CONCLUSION: Radiology report databases represent a resource from which broad descriptions of the use and results of imaging studies can be obtained. Such descriptions may be useful in departmental and hospital administration, technology assessment, cost-effectiveness studies, and health policy formulation. PMID- 9725274 TI - Liability of the moonlighting resident. PMID- 9725275 TI - Thoracic manifestations of external beam radiotherapy. PMID- 9725276 TI - Pitfalls in diagnosis of pulmonary embolism with helical CT angiography. PMID- 9725277 TI - Traumatic transection of a right aortic arch. PMID- 9725278 TI - The correlation of the radiologic extent of lung transplantation edema with pulmonary oxygenation. AB - OBJECTIVE: The study set out to evaluate the relationship between the efficiency of pulmonary oxygenation and the extent of the reimplantation response as revealed on chest radiography after bilateral lung transplantation. MATERIALS AND METHODS: Postoperative chest radiographs of 31 patients who had undergone bilateral lung transplantation were evaluated for the extent of the reimplantation response. For each patient, the contemporaneous oxygenation indexes (partial pressure of oxygen in arterial blood divided by fraction of inspired oxygen) were calculated and correlated with a radiographic score produced from the evaluation of chest radiographs. RESULTS: The method of evaluating chest radiographs for the extent of the reimplantation response was shown to be reproducible. Although mean oxygenation indexes were found to decrease with increasing radiographic scores, this trend was not statistically significant. CONCLUSION: Although the extent of the reimplantation response on the early postoperative chest radiography inversely correlated with the oxygenation efficiency of the transplanted lungs, this finding was not statistically significant. PMID- 9725279 TI - Mediastinal teratoma: CT differentiation of ruptured and unruptured tumors. AB - OBJECTIVE: The purpose of this study was to differentiate ruptured from unruptured mediastinal teratomas using CT. MATERIALS AND METHODS; CT findings in 17 cases of surgically resected mediastinal teratomas were reviewed retrospectively. Preoperative rupture was found in seven patients during surgery. We compared the clinical symptoms and CT findings of ruptured tumors with those of unruptured tumors. On CT, we evaluated size, wall thickness, location of the mass, presence or absence of internal septation, homogeneity of the internal components of each compartment, calcification or fat within the mass, and ancillary findings in adjacent structures. RESULTS: Severe symptoms (chest pain or hemoptysis) were more commonly found in ruptured (71%) than in unruptured tumors. All ruptured mediastinal teratomas had a tendency to display inhomogeneity of the internal components, whereas 90% of unruptured masses showed homogeneous densities of internal components in each compartment of the mass. Ancillary CT findings in ruptured tumors included fat-containing masses in adjacent lung parenchyma in two patients, consolidation or atelectasis in the adjacent lung in three patients, pericardial effusion in one patient, and pleural effusion in four patients. CONCLUSION: In cases of mediastinal teratoma, CT findings of inhomogeneity of the internal components and changes in the adjacent lung parenchyma, pleura, or pericardium can be used as signs of tumor rupture. PMID- 9725280 TI - The retained intrapericardial sponge: value of the lateral chest radiograph. AB - OBJECTIVE: Our objective was to identify reasons for the difficulty in diagnosing retained intrapericardial sponges and to determine ways to improve diagnostic accuracy. CONCLUSION: All three intrapericardially retained sponges were in the posterior pericardium, a region not visible to the surgeon. Radiographic detection of the sponges on standard anteroposterior projections is difficult because of exposure factors, other confusing linear markers, and metallic densities such as sternal sutures. However, knowledge of the typical location of a lost sponge and use of lateral radiographic projections may aid in early detection of this rare complication. PMID- 9725282 TI - Solitary peripheral papilloma of the breast: a radiologic-pathologic correlation of a benign lesion that may mimic breast cancer on mammography. AB - OBJECTIVE: The objective of this study was to describe the mammographic appearance with pathologic correlation of solitary peripheral papillomas of the breast. CONCLUSION: Solitary peripheral papillomas of the breast are benign lesions that may present mammographic features suggestive of carcinoma. Solitary peripheral papilloma is a variant related to the solitary central duct papilloma but has a different mammographic appearance because of its location and histologic architecture. The associated risk of malignancy is unclear. PMID- 9725281 TI - Gadolinium-enhanced three-dimensional MR angiography of the aorta and peripheral arteries: evaluation of a multistation examination using two gadopentetate dimeglumine infusions. AB - OBJECTIVE: Three-dimensional gadolinium-enhanced MR angiography is a rapid and accurate method that can at times image only a limited amount of anatomy during an examination. We evaluated a technique that doubled the anatomy imaged by obtaining two separate gadolinium-enhanced MR angiograms during a single examination. MATERIALS AND METHODS: Twenty-three patients referred for MR evaluation of aortic or peripheral vascular disease underwent two successive gadolinium-enhanced three-dimensional MR angiographic examinations during a single MR examination. An injection of 15 ml of gadopentetate dimeglumine was used for the first MR angiogram, and 25 ml was used for the second MR angiogram. The angiograms were quantitatively and qualitatively evaluated to determine the effect of residual gadolinium from the initial MR angiogram on the second angiogram. RESULTS: The two studies depicted either the entire aorta to the femoral arteries (n = 10) or the distal aorta to the popliteal arteries (n = 13). The total mean gadolinium dose was 0.245 mmol/kg per patient. An average of 15 min elapsed between injections. The value of arterial signal-to-noise ratio (mean, 48.8 versus 56.4) and artery-to-vein contrast-to-noise ratio (mean, 45.5 versus 49.0) increased between the first and second angiograms, respectively. Residual gadolinium elevated the values for venous signal-to-noise ratio (mean, 2.3 versus 7.2) and background-to-muscle signal-to-noise ratio (mean, 5.5 versus 10.1) on the second MR angiogram. Qualitative evaluation by three observers showed no significant differences in diagnostic usefulness or overall image quality between the first and second MR angiograms. CONCLUSION: The use of two low-dose gadolinium-enhanced three-dimensional MR angiograms during a single examination is a feasible approach to increase anatomic coverage when performing gadolinium-enhanced three-dimensional MR angiography of the aorta and peripheral vessels. Although background enhancement is slightly elevated on the second angiogram, such enhancement does not significantly change diagnostic usefulness or overall image quality. PMID- 9725283 TI - Biopsy of breast microcalcifications using an 11-gauge directional vacuum assisted device. PMID- 9725284 TI - Prone table stereotactic breast biopsy: facilitating biopsy of posterior lesions using the arm-through-the-hole technique. PMID- 9725285 TI - Doppler waveforms of the normal and collateralized inferior mesenteric artery. AB - OBJECTIVE: Our purpose was to analyze Doppler waveform changes and the caliber of the inferior mesenteric artery as a collateral vessel in occlusive disease of the abdominal aorta or its main branches. SUBJECTS AND METHODS: Thirty-three patients were examined in three groups according to the location of their occlusive disease (group 1 [n = 5], occlusion of the celiac and superior mesenteric arteries; group 2 [n = 9], occlusion of the iliac artery; and group 3 [n = 19], occlusion of the abdominal aorta distal to the renal arteries). The main truncus of the inferior mesenteric artery was evaluated along its longitudinal axis using color duplex Doppler sonography. Peak systolic velocity, end-diastolic velocity, mean velocity, resistive index, and pulsatility index were determined from the Doppler spectrum. The inner diameter and cross-sectional area of the inferior mesenteric artery were measured, and blood flow volume was calculated. The data obtained from the three groups were compared with data from a control group (n = 24). RESULTS: In all three patient groups, the mean blood flow volume and the mean flow velocities were significantly higher, the mean pulsatility index was significantly lower, and the mean diameter of the vessel was significantly larger than in the control group. The blood flow volume in patients with aortic occlusion was significantly lower than that in patients with superior mesenteric artery occlusion. In the patients with iliac artery occlusion, the mean resistive index was not significantly different from that in the control group. CONCLUSION: An increase in blood flow volume and the presence of a monophasic waveform indicate increased collateral function of the inferior mesenteric artery. However, blood flow volume in patients with aortic occlusion does not increase as high as in patients with superior mesenteric artery occlusion, and a monophasic waveform is not a distinctive finding in iliac artery occlusion. PMID- 9725286 TI - Evaluation with Doppler sonography of mesenteric blood flow in celiac disease. AB - OBJECTIVE: The aim of this study was to investigate with Doppler sonography the variations of resistance in the superior mesenteric artery, both at fasting and in the postprandial state, in patients with celiac disease. SUBJECTS AND METHODS: Twenty-five patients with celiac disease (20 women, five men; mean age, 30 +/- 7 years) and 10 healthy volunteers (seven women, three men; mean age, 28 +/- 6 years) were examined with Doppler sonography. Nineteen patients were untreated (no dietary restrictions) and six patients were treated with a gluten-free diet at the time of the examination. Imaging was performed at both fasting and 15 min after an 1890-kJ meal. We introduced a parameter called "resistive difference," defined as the mathematic difference between the resistive index measured at fasting (highest value) and that measured at 15 min after the meal (lowest value) as a way to express the postprandial resistive change in the superior mesenteric artery. RESULTS: Untreated patients with flat mucosa showed a resistive difference of 0.03 +/- 0.05, followed by untreated patients with mucosal subatrophy (0.05 +/- 0.04), treated patients (0.09 +/- 0.02), and healthy volunteers (0.12 +/- 0.04). A statistically significant difference was noticed between the resistive difference of healthy volunteers and both those of the untreated patients with subatrophy (p = .016) and of the patients with complete atrophy (p = .011), as well as between the resistive difference of the treated patients and both those of the untreated patients with subatrophy (p = .021) and of the patients with complete atrophy (p = .020). CONCLUSION: We believe that Doppler measurement of resistive difference in the superior mesenteric artery can be an effective way to express severity of celiac disease and to document its regression after diet therapy. PMID- 9725287 TI - Use of self-expanding metallic stents in the management of obstruction of the sigmoid colon. AB - OBJECTIVE: The purpose of our study was to evaluate the clinical efficacy of a new self-expanding metallic endoprosthesis in the management of distal colonic obstruction in seven patients. CONCLUSION. The Wallstent enteral endoprosthesis is safe and effective in relieving obstruction in patients with resectable colonic tumors. Once in place, the Wallstent permits planned elective surgery and avoids a temporary stoma. In addition, the Wallstent can palliate patients with obstruction due to advanced colonic neoplasms. The results of our preliminary study are promising and show a low incidence of complications. PMID- 9725288 TI - CT of colonic malakoplakia in a patient with AIDS. PMID- 9725289 TI - Midgut volvulus: a rare cause of acute abdomen in an adult patient. PMID- 9725290 TI - Using triphasic helical CT to detect focal hepatic lesions in patients with neoplasms. AB - OBJECTIVE: Our purpose was to determine the value of triphasic helical CT (unenhanced, hepatic arterial, and portal venous phases) in the detection and characterization of focal hepatic lesions due to hepatomas or metastases. MATERIALS AND METHODS: One hundred two patients with known or suspected hepatomas or liver metastases underwent triphasic CT. The number and conspicuity of lesions were evaluated on each phase. RESULTS: Five hundred eighty-four lesions were detected in 102 patients. Patients with hypovascular malignancies had more lesions detected on the portal venous phase with increased conspicuity than on the other phases. Patients with hypervascular malignancies had lesions best detected on the hepatic arterial phase, which revealed small lesions that were not seen on the other phases in seven (21%) of the 33 patients with hypervascular metastases and hepatomas. No lesions were detected on the unenhanced phase that were not seen on the other phases. However, arterial phase images introduced new diagnostic dilemmas because not all lesions seen on the arterial phase alone were caused by hepatomas or metastases, even in patients with known malignancies; several lesions represented benign abnormalities that included focal nodular hyperplasia. CONCLUSION: The unenhanced phase is not routinely necessary for the detection of metastases or hepatomas. Hypovascular malignancies are best evaluated during the portal venous phase. Small lesions due to hypervascular metastases and hepatomas are best evaluated and may be detected only during the hepatic arterial phase, which should be used routinely in these patients. New dilemmas may develop from the increased sensitivity of the hepatic arterial phase for lesions. However, the hepatic arterial phase is of limited value with hypovascular malignancies. PMID- 9725291 TI - Multiphasic helical CT in diagnosis and staging of hilar cholangiocarcinoma. AB - OBJECTIVE: The purpose of our study was to assess the potential of thin-section multiphasic helical CT in diagnosis and staging of hilar cholangiocarcinomas. SUBJECTS AND METHODS: Identically collimated helical CT studies were performed before and during the hepatic artery dominant phase and during the portal vein dominant phase of contrast enhancement in 29 consecutive patients with proven hilar cholangiocarcinomas. Differences in attenuation between the tumor and the liver were calculated in each case by subtracting the average attenuation of the tumor from that of the liver. A four-point scale termed a "lesion conspicuity score" was used to determine rates of tumor detection. CT findings were correlated with surgically assessed extent of tumor, histologic findings, or both in all cases. RESULTS: Ten (34%) of the 29 hilar cholangiocarcinomas were detected on unenhanced images. All hilar cholangiocarcinomas (100%) were seen on hepatic artery dominant phase scans, and 25 (86%) of 29 hilar cholangiocarcinomas were seen on portal vein dominant phase scans, regardless of the morphologic appearance. An infiltrating stenotic lesion was found in 17 (59%) of 29 patients, an exophytic hilar lesion was found in 11 patients (38%), and one patient (3%) had an intraluminal polypoid lesion. Mean differences in enhancement between infiltrating stenotic lesions and the liver were significantly greater on hepatic artery dominant phase scans (28 +/- 10 H) than on portal vein dominant phase scans (10 +/- 8 H), whereas the mean difference in enhancement between the exophytic lesions and the liver was statistically greater during the portal vein dominant phase (p < .01). Two of the hilar cholangiocarcinomas were resectable at surgery, and 18 were not. The overall accuracy of helical CT for assessing resectability was 60%. In 10 (56%) of 18 patients, unresectable disease was correctly diagnosed with helical CT (sensitivity, 56%). Eight (44%) of 18 patients considered to have resectable tumors with helical CT had unresectable tumors at surgery. A resectable tumor was correctly diagnosed in two patients with helical CT. CONCLUSION: Multiphasic helical CT can be used to detect and classify hilar cholangiocarcinomas. However, the exact proximal tumor extent along bile ducts tends to be underestimated with helical CT; therefore, helical CT is inaccurate for determining resectability. PMID- 9725292 TI - Diagnostic criteria for fatty infiltration of the liver on contrast-enhanced helical CT. AB - OBJECTIVE: The purpose of the study was to develop quantitative and qualitative criteria for diagnosing fatty liver on contrast-enhanced helical CT. SUBJECTS AND METHODS: Differential liver-spleen attenuation was evaluated between 80 and 120 sec after injection in 76 patients who underwent contrast-enhanced helical CT. Unenhanced CT images had earlier established fatty liver when the liver minus spleen attenuation difference was less than or equal to -10 H (n = 18). Four observers who had not seen the unenhanced images used contrast-enhanced CT images to assess the presence of fatty liver on a five-point Likert scale, the presence of geographic areas spared from fatty infiltration, and the relative liver-spleen attenuation. The diagnostic accuracies of various imaging criteria were compared using McNemar's chi-square test (for sensitivity and specificity) and analysis of receiver operating characteristic curves. RESULTS: Sensitivity, specificity, and receiver operating characteristic curve areas for observers' qualitative judgments were 54%, 95%, and .91, respectively; for quantitative differential liver-spleen attenuation (80-100 sec; -20.5 H discriminatory value), the values were 86%, 87%, and .94, respectively; and for quantitative differential liver spleen attenuation (101-120 sec; -18.5 H discriminatory value), the values were 93%, 93%, and .98, respectively. Differential liver-spleen attenuation was time dependent; overlap was noted between healthy subjects and patients with fatty liver. Qualitatively, geographic sparing was highly specific (94%) for fatty liver, whereas liver attenuation greater than or equal to spleen attenuation excluded fatty liver in all but one case. CONCLUSION: Although quantitative and qualitative criteria for diagnosing fatty liver on helical CT can be determined, they are protocol-specific. Limited unenhanced hepatic CT remains the optimal technique for detection of fatty infiltration of the liver. PMID- 9725293 TI - Gadolinium-enhanced MR imaging of traumatic hepatic injury. AB - OBJECTIVE: Our objective was to evaluate the use of contrast-enhanced MR imaging in the examination of a limited number of patients with hepatic trauma who had also undergone CT. CONCLUSION: The conspicuity of most lesions was better on contrast-enhanced MR imaging than on unenhanced MR imaging. Also, conspicuity on contrast-enhanced MR imaging was at least subjectively equivalent to that on CT. PMID- 9725294 TI - Pelvic extension of retroperitoneal fluid: analysis in vivo. AB - OBJECTIVE: The purposes of this study were to describe the pathway of fluid flow from the retroperitoneal space into the pelvic extraperitoneal space on CT in vivo, to clarify the relation between its occurrence and the site or amount of retroperitoneal fluid, and to delineate the anatomic relation between the retroperitoneal spaces and the pelvic extraperitoneal space. MATERIALS AND METHODS: We reviewed the CT scans of 37 patients with retroperitoneal fluid collections. Patients who had undergone pelvic laparotomy and patients who had either fascial thickening alone or fluid within muscle (such as the psoas muscle or iliac muscle) alone were excluded. RESULTS: Fluid extension into the pelvic extraperitoneal space was seen in six patients (16%). Extension by the infrarenal extraperitoneal space was seen in all six of these patients, but extension by properitoneal fat was seen in only one of the six patients. In patients with large amounts of fluid in the infrarenal extraperitoneal space, we frequently saw extension into the pelvic extraperitoneal space. Extension of pancreatic fluid into the infrarenal extraperitoneal space occurred in only 15% of the 37 patients. However, it occurred in both patients with ruptured abdominal aortic aneurysms. Three pathways from the infrarenal extraperitoneal space into the pelvic extraperitoneal space were seen: extension dorsally medial to the iliac vessels (n = 6), extension dorsally lateral to the iliac vessels (n = 1), and extension medially into the prevesical space (n = 2). Coexistence of two of these three pathways was seen in three patients. CONCLUSION: In vivo, extension of retroperitoneal fluid into the pelvic extraperitoneal space is not rare and occurs more often by the infrarenal extraperitoneal space than by properitoneal fat. Extension of retroperitoneal fluid to the infrarenal extraperitoneal space can be attributed less frequently to sources distant to the pelvic cavity such as pancreatic fluid. Such extension often derives from sources that can produce large amounts of retroperitoneal fluid such as ruptured abdominal aortic aneurysms. Of the three pathways from the infrarenal extraperitoneal space to the pelvic extraperitoneal space, dorsal extension medial to the iliac vessels is the most common, and multiple pathways often coexist. PMID- 9725295 TI - Active arterial contrast extravasation on helical CT of the abdomen, pelvis, and chest. PMID- 9725296 TI - CT appearance of primary papillary serous carcinoma of the peritoneum. AB - OBJECTIVE: Our objective was to describe the CT characteristics of primary papillary serous carcinoma of the peritoneum. CONCLUSION: The presence of peritoneal masses, extensive omental calcification, and the absence of an ovarian mass on CT--particularly in postmenopausal women--is highly suggestive of primary papillary serous carcinoma of the peritoneum and should alert the radiologist to the possibility of this diagnosis. PMID- 9725297 TI - Osteosarcoma of the abdominal wall with spontaneous regression of lung metastases. PMID- 9725298 TI - Negative predictive value of imaging-guided abdominal biopsy results: cytologic classification and implications for patient management. AB - OBJECTIVE: Our purpose was to assess the negative predictive value of imaging guided abdominal biopsy results and correlate it with cytology classification, lesion size, needle gauge, and cancer history. MATERIALS AND METHODS: A retrospective study was performed of 100 patients with proven diagnoses who had undergone imaging-guided abdominal biopsies showing no cells that were malignant or suspicious for malignancy. Specimens were classified as normal or benign, nondiagnostic, or atypical. Negative predictive value was calculated for each cytologic category, lesion size, needle gauge, and cancer history. Logistic regression analysis was performed to allow us to identify predictors of false negative results. RESULTS: Overall negative predictive value was 67%. Other negative predictive values were normal or benign result, 78%; nondiagnostic result, 66%; and atypical result, 29%. Negative predictive value was greater when the lesion was large (> or =3 cm) (p = .031). Logistic regression analysis allowed us to predict a 9.3% chance of a false-negative result for a specimen of normal or benign cytology that was taken from a large lesion in a patient with no cancer history but an 87% chance of a false-negative when a specimen of atypical cytology was taken from a small (<3 cm) lesion in a patient with a cancer history. CONCLUSION: Imaging-guided abdominal biopsy specimens containing atypical cells should be viewed with caution. In patients without cancer, if a lesion is large and the specimen contains normal target organ or benign cells, the likelihood of a false-negative result may be low enough that imaging surveillance at appropriate intervals may be sufficient. PMID- 9725299 TI - Extravisceral masses in the peritoneal cavity: sonographically guided biopsies in 52 patients. AB - OBJECTIVE: Our objective was to determine the effectiveness of sonographically guided biopsies of extravisceral masses (masses outside the solid organs) in the peritoneal cavity. MATERIALS AND METHODS: We retrospectively reviewed the results of sonographically guided biopsies of extravisceral masses found in the peritoneal cavity of 52 patients (age range, 25-90 years old; mean age, 52 years) from June 1990 to December 1996. Fifty-one patients underwent biopsy through the abdominal wall, and one patient underwent transvaginal biopsy. Sonographic guidance was obtained using 3.5- to 7.0-MHz vector probes. The size, depth, and sonographic characteristics of the mass and the type of biopsy (aspirate versus core) were determined for all lesions. Pathology reports and clinical courses were reviewed. RESULTS: Placement of the biopsy needle within the lesion was successful in all patients. The mean depth from skin surface to lesion was significantly less (p < .0001) when shown by sonography (2.4 cm) than when shown by CT (3.8 cm). Biopsy results were true-positive for malignancy in 37 patients (no false-positives), true-negative for benign masses in 10 patients, and false negative for malignancy in three patients (sensitivity, 93%; specificity, 100%; accuracy, 94%). Nondiagnostic samples were obtained in two patients (4%). Treatment was based on diagnostic biopsy results in 43 patients (86%). CONCLUSION: Sonography is an effective alternative to CT in guiding biopsy of extravisceral masses in the peritoneal cavity. PMID- 9725300 TI - A biopsy compression device for use in cross-sectional or fluoroscopic imaging. PMID- 9725301 TI - Preoperative staging of cervical carcinoma: phased array coil fast spin-echo versus body coil spin-echo T2-weighted MR imaging. AB - OBJECTIVE: This study was performed to compare the diagnostic efficacy of MR imaging in the preoperative evaluation of invasive cervical cancer using the pelvic phased array coil in combination with fast spin-echo T2-weighted imaging and the body coil in combination with conventional spin-echo T2-weighted imaging. MATERIALS AND METHODS: Ninety-four women (22-68 years old) with invasive cervical cancer underwent MR imaging (at 1.5 T) using a body coil conventional spin-echo protocol (n = 62) or a phased array coil fast spin-echo protocol (n = 32). Imaging preceded surgery by no more than 5 weeks. MR images were evaluated for tumor size, local stage, and nodal metastasis using surgical pathology as the standard of reference. RESULTS: Overall staging accuracy for the body coil conventional spin-echo protocol (89%) was not significantly different from that of the phased array coil fast spin-echo protocol (91%). Both techniques also achieved similar accuracy in diagnosing parametrial invasion (95% versus 94%) and lymph node metastases (85% versus 91%) and in tumor sizing (correlation coefficient, .93 versus .94). CONCLUSION: In the preoperative staging of cervical carcinoma by MR imaging, both the newer (phased array coil fast spin-echo protocol) and the older (body coil conventional spin-echo protocol) techniques achieved similarly high accuracies in local staging, assessment of parametrial invasion, and evaluation of tumor size. Decreased imaging time and increased image resolution are advantages of the newer technique, although in our series they did not increase staging accuracy. PMID- 9725302 TI - Diagnostic sonography of HIV-associated nephropathy: new observations and clinical correlation. AB - OBJECTIVE: HIV-associated nephropathy is an important cause of morbidity that is characterized clinically by uremia and proteinuria and histologically by focal segmental glomerulosclerosis. In the largest series yet analyzed to our knowledge, we describe new sonographic findings and record the prevalence of other findings. We review the sonographic findings in a large group of HIV infected patients. MATERIALS AND METHODS: Seventy-six consecutive HIV-infected patients underwent renal sonography. Abnormalities seen on sonography were recorded. RESULTS: Of 152 kidneys imaged, sonography showed that 30 kidneys (20%) were enlarged. Abnormal echogenicity was present in 136 kidneys (89%). Eighty-one kidneys (53%) were globular; 58 (38%) had decreased corticomedullary definition; 74 (49%) had decreased renal sinus fat; and 66 (43%) had heterogeneous parenchyma, some with echogenic striations. CONCLUSION: Our data reveal several sonographic abnormalities that have not previously been described: decreased corticomedullary definition, decreased renal sinus fat, parenchymal heterogeneity, and globular renal configuration. These new findings were found mainly in patients with advanced HIV infection. PMID- 9725303 TI - Nonopaque crystal deposition causing ureteric obstruction in patients with HIV undergoing indinavir therapy. AB - OBJECTIVE: We describe the unique CT features of ureteric calculi in six HIV infected patients receiving indinavir, the most commonly used HIV protease inhibitor, which is associated with an increased incidence of urolithiasis. CONCLUSION: Ureteric obstruction caused by precipitated indinavir crystals may be difficult to diagnose with unenhanced CT. The calculi are not opaque, and secondary signs of obstruction may be absent or minimal and should be sought carefully. Images may need to be obtained using i.v. contrast material to enable diagnosis of ureteric stones or obstruction in patients with HIV infection who receive indinavir therapy. PMID- 9725304 TI - Extraadrenal myelolipoma: MR imaging findings. PMID- 9725305 TI - Lymphoproliferative disorders: CT findings in immunocompromised children. AB - OBJECTIVE: The objective was to evaluate the CT imaging appearance, distribution of disease, type of immunocompromised state, and outcome of children with Epstein Barr virus-induced lymphoproliferative disorders. MATERIALS AND METHODS: Medical records and imaging studies (from four tertiary children's medical centers) were reviewed for pathologically proven cases of lymphoproliferative disorders in patients less than 20 years old. Trends between the CT imaging appearance, distribution, and type of immunocompromised state and prognosis were noted and analyzed with Fisher's exact test. RESULTS: Twenty-seven cases were identified (mean age, 7 years 8 months). Eighteen children had undergone solid organ transplantation (heart, n = 9; liver, n = 7; kidney, n = 2), and four had undergone bone marrow transplantation. Five patients had primary immunodeficiencies. The CT appearance of lymphoproliferative disorders varied and included lymphadenopathy, focal mass or masses, and diffuse infiltration and enlargement of organs without focal mass. The distribution of disease included abdomen (n = 17), chest (n = 10), neck (n = 8), and brain (n = 1). In eight of nine heart transplant recipients, the disease predominantly involved the chest and neck, whereas in all seven liver transplant recipients, the disease was isolated to the abdomen (p = .001). The overall mortality rate of 44% was less related to anatomic extent (multiorgan, 46%; localized, 43%) than to type of immune dysfunction (p = .001): bone marrow transplantation (100%), primary immunodeficiency (80%), heart transplantation (55%), liver transplantation (0%), and kidney transplantation (0%). CONCLUSION: Lymphoproliferative disorders in children had a variable distribution, imaging appearance, and outcome. However, in recipients of solid organ transplants, the disease tended to occur in the anatomic region of the transplant. Mortality rates were more closely related to the type of underlying immune dysfunction than to distribution of disease. PMID- 9725306 TI - Nonmalignant renal disease in pediatric patients with Beckwith-Wiedemann syndrome. AB - OBJECTIVE: The objective of this retrospective review was to determine the incidence and spectrum of nonmalignant renal disease in patients with Beckwith Wiedemann syndrome. MATERIALS AND METHODS: Patient records were obtained from the Beckwith-Wiedemann Registry of the National Cancer Institute. Imaging findings and medical records of 152 neonates, infants, children, and adults with Beckwith Wiedemann syndrome (age range, 1 day to 30 years old; median age, 1 year 3 months old) were retrospectively reviewed by three radiologists. Available pathologic material also was reviewed. RESULTS: Thirty-eight (25%) of 152 patients with Beckwith-Wiedemann syndrome had 45 nonmalignant renal abnormalities, including medullary renal cysts (n = 19, 13%), caliceal diverticula (n = 2, 1%), hydronephrosis (n = 18, 12%), and nephrolithiasis (n = 6, 4%). Thirty-three (87%) of the 38 patients with nonmalignant renal disease were asymptomatic. Clinical manifestations of the remaining five patients included urinary tract infections (n = 4) and flank pain due to obstructive stone disease (n = 1). Nonmalignant renal disease was mistaken for Wilms' tumor in two patients, resulting in unnecessary nephrectomies. Seven children (18%) had Wilms' tumor and nonmalignant renal disease. CONCLUSION: Nonmalignant renal abnormalities occur in approximately 25% of patients with Beckwith-Wiedemann syndrome but are generally asymptomatic. Nonmalignant renal abnormalities should be considered in the differential diagnosis of a mass revealed during screening sonography of a patient with Beckwith-Wiedemann syndrome to avoid unnecessary surgery. PMID- 9725307 TI - Sonography of patellar abnormalities in children. AB - OBJECTIVE: We describe our experience using sonography to evaluate the patella and patellofemoral joint in three children with congenital abnormalities of the extensor mechanism of the lower extremity. CONCLUSION: Sonography can be used to evaluate the extensor mechanism and unossified patella in young children. Advantages of sonography over MR imaging are no need for sedation, speed of examination and ability to compare with the contralateral knee, low cost, and the sonographer's ability to dynamically evaluate the extensor mechanism. PMID- 9725308 TI - Color Doppler sonography of patellar tendinosis. PMID- 9725310 TI - Extended field-of-view sonography in musculoskeletal disorders. PMID- 9725309 TI - Sonography of Achilles tendon xanthomas in patients with heterozygous familial hypercholesterolemia. AB - OBJECTIVE: Xanthomas are an essential diagnostic criteria of familial hypercholesterolemia. The objective of this study was to determine if xanthomas of the Achilles tendon can be revealed on sonography when the condition is clinically unsuspected in patients with heterozygous familial hypercholesterolemia. SUBJECTS AND METHODS: Ninety-four patients (52 females, 42 males; 12-73 years old; mean age, 44 years) with a proven diagnosis of heterozygous familial hypercholesterolemia were studied. Achilles tendons of these patients were clinically evaluated and examined with sonography. Size and echo structure of the Achilles tendons were categorized as normal, having hypoechoic nodules (grade 1), or diffusely hypoechoic with a heterogeneous echo structure (grade 2). RESULTS: Sixty-two patients had positive physical examination findings for Achilles tendon xanthomas. Fifty-seven (92%) of these patients had abnormally large tendons (> or =7.1 mm) on sonography. Grade 1 or grade 2 echo structure compatible with xanthomatosis was found in at least one Achilles tendon of all 62 patients. Of the 94 patients in the study group, 32 patients had negative or indeterminate physical examination findings for Achilles tendon xanthomas. Sonography showed that two (6%) of these patients had an enlarged (> or =7.1 mm) Achilles tendon. Grade 1 or grade 2 echo structure compatible with xanthomatosis was found in 26 (81%) of these 32 patients. CONCLUSION: Sonography is significantly more sensitive than physical examination for the detection of Achilles tendon xanthomas in patients with heterozygous familial hypercholesterolemia and normal-sized Achilles tendons. Our study suggests that sonography may play an important role in the early diagnosis of heterozygous familial hypercholesterolemia. PMID- 9725311 TI - Medicine in American art. Night call. PMID- 9725312 TI - Accuracy of CT-guided needle biopsy of musculoskeletal neoplasms. AB - OBJECTIVE: The purpose of our study was to assess the accuracy of CT-guided biopsy of musculoskeletal neoplasms with respect to technique, anatomic site, and histology. MATERIALS AND METHODS: During a 3-year period (January 1992 to December 1994), 176 core needle biopsies and 45 fine-needle aspirations were performed under CT guidance on patients with musculoskeletal neoplasms. To assess the accuracy of these procedures, we compared the diagnosis at biopsy with the final diagnosis as determined at the time of definitive treatment of the lesion. All biopsy findings were categorized as a primary malignancy (excluding round cell lesions), round cell lesion, local recurrence, or metastatic carcinoma. In addition, each lesion was analyzed according to which biopsy technique was used, whether frozen tissue section or rapid cytologic evaluation was used, and at which anatomic site the mass was found. RESULTS: The accuracy for needle biopsy was 93% and that for fine-needle aspiration was 80%. The complication rate for both techniques was less than 1%. Accuracy rates for the four categories of primary malignancy, round cell lesion, local recurrence, and metastatic carcinoma were 87%, 75%, 94%, and 100%, respectively. The mismatch rates were similar in soft-tissue lesions (5/52) and bone lesions (16/169). Diminished accuracy was associated with round cell lesions (20%) and lesions located in the spine or the perivertebral region (20%). Nondiagnostic and insufficient specimens were found in 18 (8%) of the 221 patients. CONCLUSION. CT-guided biopsy of musculoskeletal malignancies is a safe and effective procedure if performed by a team of clinicians, pathologists, and radiologists who possess subspecialty expertise. PMID- 9725313 TI - Tear of the posterior shoulder stabilizers after posterior dislocation: MR imaging and MR arthrographic findings with arthroscopic correlation. AB - OBJECTIVE: The purpose of this article is to describe the findings on MR imaging and MR arthrography in posterior capsular tear and teres minor muscle injury after posterior dislocation. We also correlate MR imaging with the arthroscopic findings and present treatment options for these patients. CONCLUSION: MR imaging is helpful in diagnosing abnormalities caused by posterior dislocation injuries and in directing therapy. Teres minor muscle and capsular injuries may occur without the typical reverse Bankart lesion. Isolated teres minor muscle tears seen on MR imaging after posterior dislocation injury may cause pain. However, no consensus exists as to whether the lesions seen on MR images in these patients should be treated surgically or conservatively. PMID- 9725314 TI - MR imaging of the shoulder after an impingement test: how long to wait. AB - OBJECTIVE: The purpose of this study was to determine the appropriate minimum waiting time between an impingement test with subacromial injection and subsequent MR imaging to avoid misinterpretation if the injected fluid is still present. CONCLUSION: MR imaging should be delayed a minimum of 24 hr after a subacromial injection. Fluid in the subacromial space 24 hr after subacromial injection is unrelated to an impingement test. PMID- 9725315 TI - Long bone surface osteomas: imaging features that may help avoid unnecessary biopsies. AB - OBJECTIVE: Our purpose is to show that a combination of imaging techniques and periodic radiologic follow-up offers an alternative to biopsy in certain patients with long bone surface osteomas. CONCLUSION: Asymptomatic lesions that are consistent with osteoma on a combination of imaging studies can be followed up clinically and radiographically, allowing patients to avoid unnecessary biopsies. PMID- 9725316 TI - Muscle denervation patterns in upper limb nerve injuries: MR imaging findings and anatomic basis. PMID- 9725317 TI - Coccidioidal spondylitis: MR findings in 15 patients. AB - OBJECTIVE: MR imaging studies of 15 patients with documented vertebral column coccidioidomycosis infection were retrospectively reviewed to determine the MR imaging features of coccidioidal spondylitis. CONCLUSION: On MR imaging, coccidioidal spondylitis may be unifocal or multifocal. Involvement of an intervertebral disk, vertebral body marrow, and adjacent epidural and soft tissue is generally seen. PMID- 9725318 TI - Cerebral infarction: time course of signal intensity changes on diffusion weighted MR images. AB - OBJECTIVE: The objective of this study was to determine the time course of signal intensity changes on diffusion-weighted MR images after cerebral infarction. MATERIALS AND METHODS: Echoplanar diffusion-weighted MR images were obtained at 1.5 T in 212 patients referred for suspected cerebral infarction over a 6-month period. Of those patients, 85 met strict criteria for inclusion in this study: final clinical diagnosis of stroke, reliable timing of clinical ictus by history, and neurologic symptoms persisting longer than 48 hr after onset. Using adjacent or contralateral normal brain for comparison, diffusion-weighted images were visually analyzed retrospectively to evaluate for abnormalities in signal intensity. Because three patients were scanned on two occasions and five patients had two anatomically separable infarctions, 93 reliably dated brain lesions were analyzed. RESULTS: Diffusion-weighted images showed abnormal findings in 13 (100%) of 13 lesions less than 1 day old, 46 (96%) of 48 lesions 1-4 days old, 16 (94%) of 17 lesions 5-9 days old, three (60%) of five lesions 10-14 days old, and zero (0%) of 10 lesions more than 14 days old. CONCLUSION: Abnormal signal intensity was present on all diffusion-weighted MR studies obtained in patients within 24 hr of acute cerebral infarction and in up to 94% of patients scanned during the first 2 weeks after ictus. The percentage of abnormal diffusion studies declined with time, and no signal intensity abnormality was seen in stroke patients scanned more than 2 weeks after symptom onset. PMID- 9725319 TI - MR imaging of hemorrhagic brain lesions: a comparison of dual-echo gradient- and spin-echo and fast spin-echo techniques. AB - OBJECTIVE: Our objective was to assess the usefulness of the dual-echo gradient- and spin-echo (GRASE) technique in revealing acute hemorrhagic brain lesions and compare GRASE and fast spin-echo techniques for revealing acute hemorrhagic lesions and image artifacts. MATERIALS AND METHODS: Thirty-two consecutive patients with acute intracranial hemorrhage underwent dual-echo GRASE (TEeff1/TEeff2, 35/85) and fast spin-echo (25/110) imaging. The techniques were matched for TR (3032 msec), spatial resolution, and acquisition time. Two neuroradiologists reviewed the images independently, documenting the number, size (<1, >1, or 1 cm in diameter), location, and signal characteristics (hypointense versus hyperintense compared with brain) of detectable lesions. These observers also compared matched T2- and proton density-weighted GRASE and fast spin-echo images for paramagnetic lesion conspicuity, diamagnetic susceptibility artifacts, chemical shift artifacts along the phase- and frequency-encoding directions, and artifactual CSF hyperintensity in the thin curvilinear cortical sulci and the Virchow-Robin spaces on only the proton density-weighted images. RESULTS: The average number and conspicuity of dark (paramagnetic) lesions were significantly greater on GRASE than on fast spin-echo images (p < .05 and p < .001, respectively). We found no significant difference in the average number of bright lesions revealed by either technique (p > .1). Chemical shift artifacts along the phase-encoding directions were more prominent on GRASE than on fast spin-echo imaging. Chemical shift artifacts along the frequency-encoding directions and artifactual CSF hyperintensity were more prominent on fast spin-echo than on GRASE imaging. No visually apparent difference was found in the degree of diamagnetic susceptibility artifacts seen with the two techniques. CONCLUSION: Dual-echo GRASE imaging can be helpful in the examination of patients with suspected acute brain hemorrhage. PMID- 9725320 TI - Usefulness of optimized gadolinium-enhanced fast fluid-attenuated inversion recovery MR imaging in revealing lesions of the brain. AB - OBJECTIVE: The purpose of this study was to compare the contrast enhancement of lesions of the brain revealed by gadolinium-enhanced optimized fast fluid attenuated inversion recovery (FLAIR) MR imaging with that of lesions on gadolinium-enhanced optimized T1-weighted spin-echo MR imaging. SUBJECTS AND METHODS: Using computer simulations, we optimized the fast FLAIR parameters (TR, TEeff, and inversion time) and the T1-weighted spin-echo parameters (TR and TE) to provide maximum difference in signal intensity between enhancing lesions of the brain and white matter. Seventy-six consecutive patients referred for single dose gadolinium-enhanced MR imaging of the brain underwent both optimized techniques, which were matched for spatial resolution, bandwidth, and number of excitations. The gadolinium-enhanced fast FLAIR and T -weighted spin-echo MR images were evaluated independently by two observers for number and size of enhancing lesions and for the degree of gray-white matter differentiation. Contrast-to-noise ratios were measured for enhancing lesions 1.0 cm or larger in diameter using 8 x 8 pixel regions of interest in the enhancing lesions and normal white matter. RESULTS: The most revealing parameters for fast FLAIR MR imaging proved to be a TR of 1500 msec, an inversion time of 683 msec, and a TEeff of 16 msec. For T1-weighted spin-echo MR imaging, the optimized parameters were a TR of 550 msec and a TE of 16 msec. In 28 patients, we saw enhancing lesions of the brain with at least one MR imaging technique. More lesions were seen on the T1-weighted spin-echo sequence (n = 141) than on the fast FLAIR sequence (n = 94) (p < .03). Gray-white matter differentiation was significantly better on the fast FLAIR sequence (p < .001). Contrast-to-noise ratios of enhancing lesions were greater on the T1-weighted spin-echo sequence (p < .001). CONCLUSION: In this study, optimized gadolinium-enhanced conventional T1-weighted spin-echo MR imaging proved superior to gadolinium-enhanced fast FLAIR MR imaging in revealing lesions of the brain. PMID- 9725321 TI - CNS infections with free-living amebas: neuroimaging findings. AB - OBJECTIVE: The purpose of this report is to describe the clinical history, treatment, pathology, and imaging in two cases of rare CNS infection caused by free-living amebas. The Naegleria fowleri and Acanthamoeba species cause primary amebic meningoencephalitis and granulomatous amebic encephalitis, respectively. We describe the neuroimaging findings of a case involving a nonspecific cerebral edema pattern in primary amebic meningoencephalitis and a case involving focal enhancing lesions in granulomatous amebic encephalitis. CONCLUSION: Primary amebic meningoencephalitis and granulomatous amebic encephalitis have a grave prognosis and, although rare, should be considered in the differential diagnosis for patients who present with appropriate histories and imaging findings, including nonspecific brain edema on CT in primary amebic meningoencephalitis and focal punctate enhancing lesions in the posterior cranial fossa on T1-weighted MR imaging in granulomatous amebic encephalitis. PMID- 9725322 TI - MR imaging of CNS tractopathy: wallerian and transneuronal degeneration. PMID- 9725324 TI - Case 1: Horseshoe kidney with severe congenital hydronephrosis (ureteropelvic junction obstruction). PMID- 9725323 TI - Use of color and power Doppler sonography to identify feeding arteries associated with parathyroid adenomas. AB - OBJECTIVE: The objective of our study was to determine the value of using color and power Doppler sonography to reveal extrathyroidal feeding arteries in the detection of abnormal parathyroid glands. SUBJECTS AND METHODS: Forty-four patients with primary hyperparathyroidism were imaged prospectively with high resolution gray-scale, color flow, and power Doppler sonography. The presence of extrathyroidal arteries supplying the adenomas was noted. All patients underwent subsequent neck exploration. The locations of the abnormal glands were recorded. RESULTS: At surgery, 51 abnormal parathyroid glands were removed in the 44 patients. Sonography correctly revealed an adenoma in 40 of the 44 patients. Likewise, sonography revealed 42 of the 51 adenomas. Nine false-negative and two false-positive interpretations of the sonograms were made. Thus, overall sensitivity was 83%, specificity was 98%, and accuracy was 94%. Three of the false-negative interpretations were ectopic glands within the superior mediastinum. Excluding these three glands from analysis, the sensitivity for detection of adenomas within the neck was 88%, specificity was 98%, and accuracy was 95%. An extrathyroidal artery leading to a parathyroid adenoma was seen in 35 of the 42 adenomas revealed by sonography. The presence of an extrathyroidal artery leading to an adenoma was found to aid in the detection of an otherwise inconspicuous parathyroid gland in five patients, which improved sensitivity from 73% to 83%. CONCLUSION: Prominent vessels supplying parathyroid adenomas are frequently revealed by color flow and power Doppler sonography. These vessels can serve as "road maps" to abnormal parathyroid glands. PMID- 9725325 TI - Case 2: Adenocarcinoma of the female urethra. PMID- 9725326 TI - Case 3: Adrenocortical carcinoma. PMID- 9725327 TI - Case 4: Acute oliguric renal failure caused by hyperparathyroid hypercalcemic nephrocalcinosis nephropathy. PMID- 9725328 TI - Case 1: Cryptogenic organizing pneumonia. PMID- 9725329 TI - Case 2: Malignant pleural mesothelioma. PMID- 9725330 TI - Case 3: Neurofibromatosis I with metastases from a thigh neurofibrosarcoma. PMID- 9725331 TI - Case 4: Aortic pseudoaneurysm after coronary artery bypass. PMID- 9725332 TI - Case 1: TIPS-to-biliary system fistula. PMID- 9725334 TI - Case 3: Phlegmasia cerulea dolens. PMID- 9725333 TI - Case 2: Pelvic fracture with tear of the left internal pudendal artery. PMID- 9725335 TI - Case 4: Congenital arteriovenous malformation of the kidney. PMID- 9725336 TI - Case 1: Afferent loop syndrome. PMID- 9725337 TI - Case 2: Pseudomembranous colitis. PMID- 9725338 TI - Case 3: T-cell lymphoma of the small bowel developing in long-standing celiac disease (nontropical sprue). PMID- 9725339 TI - Case 4: Cecal volvulus. PMID- 9725340 TI - Case 1: Juxtacortical chondrosarcoma. PMID- 9725341 TI - Case 2: Iatrogenic soleus muscle infarct. PMID- 9725342 TI - Case 3: Aggressive osteoblastoma. PMID- 9725343 TI - Case 4: Synovial sarcoma. PMID- 9725344 TI - Case 1: Extraaxial neuroepithelial cyst involving leptomeninges and brain. PMID- 9725345 TI - Case 2: Isolated multiple sclerosis of the cervical cord. PMID- 9725346 TI - Case 3: Temporal bone rhabdomyosarcoma. PMID- 9725347 TI - Case 4: Neurocysticercosis with cervical meningeal involvement. PMID- 9725348 TI - MR imaging of fat necrosis. PMID- 9725349 TI - Computed radiology. PMID- 9725350 TI - More on the consequences of being accused of malpractice. PMID- 9725352 TI - Sonographic guidance for central venous access. PMID- 9725353 TI - Lymphomas in the lung associated with Sjogren's syndrome. PMID- 9725354 TI - Persistent hypoglossal artery. PMID- 9725355 TI - Intraventricular hemorrhage during MR imaging. PMID- 9725356 TI - Aberrant gastric venous drainage in focal fatty liver of segment IV: demonstration with sonography. PMID- 9725357 TI - Intervertebral cervical disk calcification requiring operative management in a child. PMID- 9725358 TI - Malignant lymphoma of mucosa-associated lymphoid tissue (MALT lymphoma) arising in the thymus: radiologic findings. PMID- 9725359 TI - Primary malignant fibrous histiocytoma of the liver: CT and MR findings. PMID- 9725360 TI - Early hyperenhancement of confluent hepatic fibrosis on dynamic MR imaging. PMID- 9725361 TI - Effects of cardioplegic solutions on vasoreactivity of the internal mammary artery. PMID- 9725362 TI - Nitric oxide downregulates lung macrophage inflammatory cytokine production. AB - BACKGROUND: Inflammatory cytokine production contributes to lung injury after lung ischemia reperfusion and during lung transplant rejection. Although nitric oxide has been demonstrated to reduce lung injury associated with the adult respiratory distress syndrome, it remains unknown whether the mechanism of nitric oxide's beneficial effects involves reducing lung macrophage inflammatory cytokine production. The purpose of this study was to determine whether nitric oxide downregulates lung macrophage inflammatory cytokine production. METHODS: Lung macrophages were harvested by bronchoalveolar lavage (10(6) macrophage per milliliter from normal Sprague-Dawley rats, 6 animals per group) and treated under ex vivo tissue culture conditions with the nitric oxide releasing compound S-nitoso-N-acetyl-D, L-penicillamine (0, 10(-5) 10(-4), 10(-3), 10(-2) mol/L) before induction of inflammatory cytokines with endotoxin, (50 ng/mL for 24 hours). Supernatants were assayed for inflammatory cytokine production (tumor necrosis factor alpha, interleukin-1beta) by enzyme-linked immunosorbent assay. RESULTS: Continuous nitric oxide release by S-nitoso-N-acetyl-D, L-penicillamine decreased lung macrophage tumor necrosis factor-alpha and interleukin-1beta production in a dose-dependent fashion (6 rats per group; data were analyzed for significance [p < 0.05] using two-way analysis of variance with Tukey's post-hoc correction). CONCLUSIONS: Nitric oxide decreases inflammatory cytokine production by lung macrophage. The mechanism of nitric oxide's beneficial effects may be partially attributable to decreased production of inflammatory cytokines. Nitric oxide may serve an expanded role for reducing inflammatory cytokine production during acute lung injury, ischemia-reperfusion-induced inflammation, or lung transplant rejection. PMID- 9725363 TI - Transbronchial gene transfer of endothelial nitric oxide synthase to transplanted lungs. AB - BACKGROUND: Experiments were designed to study the efficiency, distribution, and toxicity of transbronchial adenoviral-mediated transfer of endothelial constitutive nitric oxide synthase (ecNOS) gene to transplanted lungs. METHODS: Syngeneic orthotopic single-lung transplantation in the rat was performed after airway administration (300 microL, 1 x 10(9) pfu/mL) of either the ecNOS gene or the marker gene beta-Gal (control group) to donor lungs (n=4 each). After 4 days, transgene expression, inflammation, and the presence of apoptosis in the transplanted lungs were assessed by molecular, immunohistochemical, and histologic techniques. RESULTS: Gene transfer was confirmed by a positive polymerase chain reaction signal for the recombinant ecNOS gene, and recombinant messenger RNA by reverse transcription polymerase chain reaction. Positive immunohistochemical staining for ecNOS was present in more than 75% of pneumocytes only in ecNOS transduced lungs. Calcium-dependent nitric oxide synthase activity was increased in ecNOS- compared with betaGal-transduced lungs (2,139+/-756 versus 47+/-28 pmol x mg protein(-1) x h(-1); p < 0.05). Minimal to mild inflammation was observed in all transplanted lungs; fewer than 0.5% of cells in both groups were apoptotic. CONCLUSIONS: Transbronchial transfer of ecNOS gene to the transplanted lung results in transduction of pneumocytes with expression of a functionally active transgene product. PMID- 9725364 TI - Results of operation in Mycobacterium avium-intracellulare lung disease. AB - BACKGROUND: Although operation remains part of the management of Mycobacterium avium-intracellulare lung disease, few series have assessed operation in the era of better therapeutic drugs (especially clarithromycin). METHODS: From January 1, 1989, through June 30, 1997, 28 patients with M avirum-intracellulare lung disease underwent pulmonary resection. All were receiving multidrug therapy (17 of 28 were receiving clarithromycin) before and after operation. Eight patients underwent pneumonectomy (6 right, 2 left); 20 patients underwent partial resections including 18 with upper lobe lobectomies (14 right, 4 left). The most common indications for operation were medical treatment failure (15) and as part of initial therapy (9). RESULTS: Mean postoperative follow-up was 39 months. Complications occurred in 9 of 28 patients (32%), and included persistent air leak requiring surgical correction (5), early postoperative death (2), and late bronchopleural fistulae (1 patient). Twenty-three of 26 patients were known to be acid fast bacilli culture negative within 1 month of operation. Only 1 of 26 patients who survived 2 years is known to have had a relapse. CONCLUSIONS: Operation continues to play an important role in treatment of M avium intracellulare lung disease. More than 90% of patients become culture negative and remain so when they continue to receive drugs. Although morbidity is relatively high, it is manageable and the 12-month mortality in the current series was low (7%). PMID- 9725365 TI - Outcome of Medicare patients with emphysema selected for, but denied, a lung volume reduction operation. AB - BACKGROUND: Lung volume reduction operation shows promise in relieving symptoms and improving function in highly selected patients with emphysema. Withdrawal of Medicare funding for patients selected for operation by standard criteria created a matched control group with which to compare lung volume reduction recipients. METHODS: A retrospective study was done comparing 22 volume reduction candidates denied operation with 65 contemporaneous and comparable volume reduction recipients. Baseline physiologic characteristics were compared and longitudinal measures of pulmonary function were followed up for 24 months. RESULTS: Patients denied operation were similar to volume reduction recipients in all baseline measurements. Patients denied operation experienced a progressive worsening of their function, whereas volume reduction patients experienced sustained improvements. Absolute survival to date is 82% for the surgical group and 64% for the medical group. CONCLUSIONS: The improvement seen in volume reduction patients cannot be attributed to the effects of patient selection or preoperative and postoperative rehabilitation. PMID- 9725366 TI - Lung transplantation for cystic fibrosis: effective and durable therapy in a high risk group. AB - BACKGROUND: The purpose of this study was to review our experience with lung transplantation in patients with end-stage cystic fibrosis. METHODS: Eight-two patients with cystic fibrosis have undergone bilateral lung transplantation (n=76) or bilateral lower lobe transplantation (n=6) since October 1990. RESULTS: Actuarial survival for the entire cohort is 79% at 1 year and 57% at 5 years. The development of bronchiolitis obliterans syndrome is the leading cause of death after the first year. Freedom from bronchiolitis obliterans syndrome is 84% at 1 year and 51% at 3 years. Pulmonary function tests improve dramatically in recipients. There was no association between death within 1 year and recipient age, weight, graft ischemic time, cytomegalovirus seronegativity, or the presence of pan-resistant organisms. Similarly, there was no association between the development of bronchiolitis obliterans syndrome within 2 years and ischemic time, number of rejection episodes, cytomegalovirus seronegativity, or the presence of panresistant organisms. CONCLUSIONS: Despite their poor nutritional status and the presence of multiply resistant organisms, patients with cystic fibrosis can undergo bilateral lung transplantation with acceptable morbidity and mortality. PMID- 9725367 TI - Zenker's diverticulum in the elderly: is operation justified? AB - BACKGROUND: Surgical correction of symptomatic Zenker's diverticulum is effective; however, elderly symptomatic patients may be denied surgical intervention because of perceived increased risks. METHODS: To address this concern, we reviewed 75 patients (46 men and 29 women) found to have this condition during the past two decades. RESULTS: Median age was 79 years (range, 75 to 91 years). Preoperative symptoms included dysphagia in 69 patients (92%), regurgitation in 61 (81%), pneumonia in 9 (12%), halitosis in 3 (4%), and weight loss in 1 (1%). Gastroesophageal reflux symptoms were noted in 27 patients (36%). Diagnosis was made by barium swallow in 63 patients, esophagoscopy in 5, and a combination of both in 7. Surgical procedures included both diverticulectomy and myotomy in 57 patients (76%), myotomy alone in 9 (12%), diverticulopexy and myotomy in 5 (7%), and diverticulectomy alone in 4 (5%). There was no in-hospital mortality. Complications occurred in 8 patients (11%) and included esophagocutaneous fistula in 4, pneumonia and urinary tract infection in 1, and wound infection, myocardial infarction, and persistent diverticulum in 1 each. Follow-up was available in 72 patients (96%) and ranged from 8 days to 17 years (median, 3.3 years). At follow-up, 64 patients (88%) were asymptomatic and 4 (6%) were improved with minimal symptoms. The remaining 4 patients (6%) have had varying degrees of dysphagia and all have been treated with periodic esophageal dilations. CONCLUSIONS: Operation for symptomatic Zenker's diverticulum in the elderly is safe and effective and will result in resolution of symptoms and improved quality of life in most patients. PMID- 9725368 TI - Recombinant Kunitz protease inhibitor ameliorates reperfusion injury in rat lung transplantation. AB - BACKGROUND: Recombinant Kunitz protease inhibitor (rKPI-BG022) is more homologous to human Kunitz protease inhibitor than is aprotinin. Because aprotinin has been reported to inhibit free radicals, we hypothesized that rKPI would ameliorate reperfusion injury caused by free radicals. We examined its effect and the timing of administration in an in vivo rat lung transplantation model. METHODS: All lungs were flushed with low-potassium dextran-1% glucose solution and stored for 24 hours at 4 degrees C, then orthotopic left lung transplantations were performed. Rats were divided into 4 groups (n=6) as follows: group 1 served as control; in Group 2, rKPI was added to the flush solution (10 micromol/L); in group 3, rKPI (5 mg/kg) was administered intravenously to the recipient just after reperfusion; and in group 4, rKPI was added to the flush solution (10 micromol/L) and rKPI (5 mg/kg) was administered intravenously to the recipient just after reperfusion. Twenty-four hours after transplantation, the right main pulmonary artery and right main bronchus were ligated, and the rats were ventilated with 100% O2 for 5 minutes. Peak airway pressure, blood gas analysis, serum lipid peroxide level, tissue myeloperoxidase activity, and wet-dry weight ratio were measured. RESULTS: The partial oxygen tension values of group 2 were higher than those of groups 1 and 4 (groups 1, 2, and 4: 104.8+/-15.8, 245.1+/ 49.0, 101.4+/-4.5 mm Hg, respectively; p < 0.01). The partial carbon dioxide tension values of groups 3 and 4 were lower than those of group 1 (groups 1, 3, and 4: 74.5+/-5.7, 42.0+/-11.0, 46.0+/-8.4 mm Hg, respectively; p < 0.05). Peak airway pressures were lower in groups 2 and 3 than in groups 1 and 4 (groups 1, 2, 3, and 4: 22.5+/-0.5, 18.2+/-0.5, 19.2+/-0.8, 22.5+/-1.1 mm Hg; p < 0.01). Serum lipid peroxide levels in groups 2 and 3 were lower than those of groups 1 and 4 (groups 1, 2, 3, and 4: 0.793+/-0.037, 0.577+/-0.069, 0.560+/-0.029, and 0.785+/-0.053 nmol/mL, respectively; groups 2 and 3 vs group 1, and group 3 vs group 4: p < 0.01; group 2 vs group 4: p < 0.05). There were no differences in wet-dry weight ratio and tissue myeloperoxidase activity between the groups. CONCLUSION: Recombinant Kunitz protease inhibitor ameliorates reperfusion injury caused by free radicals in an in vivo rat lung transplantation model. Administration of rKPI through the flush solution and intravenous injection after reperfusion were effective separately, but the combination of the two administrations was not effective. PMID- 9725369 TI - Effect of blood transfusion on survival after esophagogastrectomy for carcinoma. AB - BACKGROUND: There is growing evidence that blood transfusion is associated with clinical factors that can lead to transfusion-induced immunosuppression. This effect can be beneficial or deleterious. METHODS: The effect of perioperative allogeneic blood transfusion on survival was studied retrospectively in 524 patients who were discharged from the hospital after esophagogastrectomy for carcinoma performed in a single unit over a 10-year period. RESULTS: The median operative blood loss for the series was 500 mL (range, 50 to 3,750 mL). Three hundred thirty-five patients (64%) received a perioperative allogeneic blood transfusion related to esophagogastrectomy, and 189 (36%) did not. The median perioperative blood transfusion administered was 900 mL (range, 300 to 12,950 mL). Perioperative allogeneic blood transfusion was associated with reduced survival for patients in stage III (p < 0.05) at 1 year, but no significant difference was found in this stage at 3 or 5 years after resection. Stage III disease accounted for 250 (48%) of the 524 patients discharged. CONCLUSIONS: Although perioperative allogeneic blood transfusion does not affect long-term survival after esophagogastrectomy for carcinoma, it does have a significant association with short-term survival in a group whose overall survival is often limited after resection. Attention should be directed toward minimizing operative blood loss and transfusing only for factors known to be clinically important, such as oxygen delivery and hemodynamics, not arbitrary hemoglobin levels. PMID- 9725370 TI - Evaluation of 18-hour lung preservation with oxygenated blood for optimal oxygen delivery. AB - BACKGROUND: Previous studies have shown that availability of oxygen during lung preservation to maintain aerobic metabolism may be essential for the optimal viability of preserved lung tissue. The purpose of this study was to evaluate lung preservation with oxygenated blood for optimal oxygen delivery to the lung graft in a rabbit model. METHODS: Eighteen excised rabbit lungs were flushed and stored for 18 hours at 10 degrees C with one of the following: Euro-Collins solution (EC; n=6), oxygenated homologous blood (OB; n=6), or low-potassium dextran solution (LPD; n=6). Poststorage lung functions were evaluated with isolated, blood-perfused lung model for 10 minutes. RESULTS: The mean oxygen tensions after reperfusion for the EC, OB, and LPD groups (47.0+/-2.8, 76.9+/ 13.1, 96.2+/-10.9 mm Hg at 10 minutes, respectively) were significantly different throughout the perfusion period (EC < OB < LPD, p < 0.05; EC < LPD, p < 0.01). Pulmonary artery pressure during the reperfusion period in the EC group (35.8+/ 4.4 mm Hg at 10 minutes) was higher than that in the OB and LPD groups (29.8+/ 4.3 and 22.4+/-2.2 mm Hg, respectively) (EC > OB, EC > LPD, p < 0.05). However, the E-selectin level in the reperfused blood in the OB group (5.04+/-0.24 ng/mL) was significantly elevated compared with that in other groups (EC, 3.56+/-0.54; LPD, 2.92+/-0.35 ng/mL, p < 0.05), which indicated enhanced neutrophil recruitment in the OB group. Comparisons of thrombomodulin and endothelin among the three groups did not reach statistical significance. CONCLUSIONS: We conclude that OB may enhance lung preservation as compared with EC solution, probably through its enriched oxygen delivery during storage and extracellular composition. However, the availability of oxygenated blood does not exceed that of LPD solution because of augmented neutrophil recruitment, which may activate neutrophil-endothelial interactions. PMID- 9725371 TI - Prospective, randomized comparison of extrapleural versus epidural analgesia for postthoracotomy pain. AB - BACKGROUND: Thoracic epidural analgesia is considered the method of choice for postthoracotomy analgesia, but it is not suitable for every patient and is associated with some risks and side effects. We therefore evaluated the effects of an extrapleural intercostal analgesia as an alternative to thoracic epidural analgesia. METHODS: In a prospective, randomized study, pain control, recovery of ventilatory function, and pulmonary complications were analyzed in patients undergoing elective lobectomy or bilobectomy. Two groups of 15 patients each were compared: one received a continuous extrapleural intercostal nerve blockade (T3 through T6) with bupivacaine through an indwelling catheter, the other was administered a combination of local anesthetics (bupivacaine) and opioid analgesics (fentanyl) through a thoracic epidural catheter. RESULTS: Both techniques were safe and highly effective in terms of pain relief and recovery of postoperative pulmonary function. However, minor differences were observed that, together with practical benefits, would favor extrapleural intercostal analgesia. CONCLUSIONS: These results led us to suggest that extrapleural intercostal analgesia might be a valuable alternative to thoracic epidural analgesia for pain control after thoracotomy and should particularly be considered in patients who do not qualify for thoracic epidural analgesia. PMID- 9725372 TI - Integrilin prevents prolonged bleeding times after cardiopulmonary bypass. AB - BACKGROUND: Cardiopulmonary bypass reduces platelet number and function, increases postoperative bleeding time, and is the major, unsolved cause of nonsurgical bleeding after open heart operations. Temporary inhibition of platelet function during cardiopulmonary bypass (platelet anesthesia) protects platelets and reduces postoperative bleeding time and bleeding. METHODS: Integrilin, a short-acting, reversible platelet glycoprotein IIb/IIIa inhibitor was studied in 28 baboons that had 60 minutes of normothermic cardiopulmonary bypass using peripheral cannulas. A control group, two groups that received different doses of Integrilin, and a group that received a combination of Integrilin and low-dose Iloprost were studied. Blood samples for platelet count, aggregation to adenosine diphosphate, beta-thromboglobulin, prothrombin fragment F1.2, thrombin-antithrombin complex, and fibrinopeptide A were obtained at seven time points. Template bleeding times were measured before and at five intervals after cardiopulmonary bypass. RESULTS: Both doses of Integrilin and the combination of Integrilin and Iloprost significantly protected platelet number, inhibited the response to adenosine diphosphate, and reduced postoperative bleeding times, but they did not reduce beta-thromboglobulin release except in the high-dose Integrilin group. Thrombin formation and activity were qualitatively, but not significantly, reduced in all treatment groups. Bleeding times were not significantly different from baseline at the time protamine was given in the combination group and 60 minutes after protamine administration in all treatment groups. CONCLUSIONS: Integrilin alone or in combination with Iloprost significantly reduces platelet activation during cardiopulmonary bypass and produces normal or near-normal bleeding times at the time protamine is given. PMID- 9725373 TI - Adenosine-enhanced ischemic preconditioning decreases infarct in the regional ischemic sheep heart. AB - BACKGROUND: Recently we have reported a myoprotective protocol, adenosine enhanced ischemic preconditioning, that extends the protection afforded by ischemic preconditioning in the isolated crystalloid-perfused heart. In this report the efficacy of adenosine-enhanced ischemic preconditioning in the in situ blood-perfused heart was investigated. METHODS: Sheep were subjected to 60 minutes of regional ischemia and 120 minutes of reperfusion. Ischemic preconditioned hearts received 5 minutes of zero flow regional ischemia and 5 minutes of reperfusion before regional ischemia. Adenosine-enhanced ischemic preconditioned hearts received a bolus injection of 10 mmol adenosine at the immediate start of ischemic preconditioning. Adenosine-treated hearts received an adenosine bolus, 10 minutes before regional ischemia. The ratio of infarct size to area at risk and mechanical function were determined. RESULTS: The infarct size to area at risk ratio in regional ischemia was 55.4%+/-2.1%. This ratio was significantly decreased with ischemic preconditioning and adenosine (22.2%+/-2.2% and 19.3%+/-1.4%, respectively; p < 0.001 versus regional ischemia) and adenosine enhanced ischemic preconditioning (8.0%+/-2.0%, p < 0.001 versus regional ischemia and ischemic preconditioning, and p < 0.01 versus adenosine). CONCLUSIONS: Adenosine-enhanced ischemic preconditioning significantly decreases infarct size in the in situ blood-perfused heart and provides superior protection compared with ischemic preconditioning. PMID- 9725374 TI - Surgical management of ascending and aortic arch disease: refined techniques with improved results. AB - BACKGROUND: Treatment of aneurysms of the ascending aorta, arch aorta, or both is surgically challenging and has traditionally carried a high hospital mortality rate. The use of refined operative techniques, including improved grafts, enhanced myocardial protection, retrograde cerebral perfusion with circulatory arrest, transesophageal echocardiography, and control of hematologic factors, has resulted in reduced hospital mortality rates. METHODS: We conducted a retrospective analysis of records of 117 consecutive patients who underwent 118 procedures between March 1987 and September 1997, for graft replacement of the ascending or transverse aortic arch with or without aortic valve reconstruction or replacement. There were 67 men (57.3%) and 50 women (42.7%). The mean age was 61.4 years (range, 16 to 81 years). Aortic abnormalities were medial degeneration in 59 patients (50.0%), dissection in 28 patients (23.7%), atherosclerosis in 16 patients (13.6%), Marfan's syndrome in 8 patients (6.8%), and other in 7 patients (5.9%). RESULTS: The ascending aorta alone was replaced in 58 patients (49.2%), ascending and arch aorta in 56 patients (47.5%), and isolated arch aorta in 4 patients (3.4%). Twenty-six patients (22.0%) required aortic valve reconstruction, 17 patients (14.4%) had separate aortic valve replacement, and 37 patients (31.4%) received a valve conduit. Overall hospital mortality rate was 3.4% (4 of 117 patients). Postoperative complications included myocardial infarction in 3 patients (2.5%), stroke in 7 patients (5.9%), pulmonary insufficiency in 22 patients (18.6%), renal insufficiency in 4 patients (3.4%), and reoperation for bleeding in 8 patients (6.8%). There were no deep sternal wound infections. Follow-up was completed for 112 (99.1%) of 113 survivors and ranged from 1 month to 10.6 years (mean, 39.5 months). Actuarial survival for patients discharged from the hospital was 87.9%+/-3.7% (standard error of the mean) at 3 years and 79.7%+/-5.8% at 6 years. CONCLUSIONS: Graft replacement of the ascending and transverse aortic arch, although technically demanding, can be performed with low hospital mortality and morbidity rates. PMID- 9725375 TI - Replacing the atherosclerotic ascending aorta is a high-risk procedure. AB - BACKGROUND: Improved techniques in cerebral and myocardial protection have made replacement of the chronically aneurysmal ascending thoracic aorta a safe and effective procedure. We hypothesized that patients with severe ascending or aortic arch atherosclerosis were at greater risk for operative complications during ascending aortic replacement because of the diffuse nature of their atherosclerotic process. METHODS: We retrospectively analyzed the records of 17 patients who received ascending aortic replacement during elective coronary artery bypass grafting (CABG) because of the intraoperative finding of severe atherosclerosis. All 17 patients underwent tube graft replacement of the ascending aorta under hypothermic circulatory arrest and retrograde cerebral perfusion before coronary artery bypass grafting. The outcomes for these patients were compared with those of a control group of 89 consecutive patients who underwent replacement for ascending thoracic aortic aneurysm. RESULTS: The hospital mortality rate for replacement of the ascending thoracic aorta for severe atherosclerosis was 23.5% (4/17) versus 2.25% (2 of 89) for the control group (p=0.006). The incidence of cerebrovascular accident in the atherosclerotic group was 17.6% (3/17) and 3.37% (3/89) for the control group (p=0.051). Nine of 17 atherosclerotic patients (52.9%) had operative morbidity. Only 20.2% (18 of 89) of the control patients had nonfatal postoperative complications. CONCLUSIONS: The severely atherosclerotic ascending aorta is a marker of diffuse atherosclerosis. Despite improved techniques of myocardial and cerebral protection, we have been unable to duplicate our success with ascending thoracic aneurysm repair. Preoperative screening of the ascending aorta by chest computed tomography may be appropriate in select high-risk patients to determine operability. PMID- 9725376 TI - Operation for acute and chronic aortic dissection: recent outcome with regard to neurologic deficit and early death. AB - BACKGROUND: We reviewed our experience in the repair of acute and chronic aortic dissection with regard to early neurologic deficit and death. METHODS: Between February 1991 and June 1996, we performed 206 operations on 195 patients for aortic dissection. Ascending or arch repair, or a combination (type A dissection) was performed on 92 of 206 patients (45%); 44 of 92 (48%) were acute dissection and 48 of 92 (52%) were chronic. Descending or thoracoabdominal repair (type B dissection) was performed on 114 of 206 patients (55%); 22 of 114 (19%) were acute and 92 of 114 (81%) were chronic. RESULTS: Among type A cases, strokes occurred in 6 of 92 patients (7%) overall; 4 of 44 (9%) were acute cases and 2 of 48 (4%) were chronic (p < 0.34). Early deaths for type A were 11 of 92 (12%) overall; 9 of 44 (20%) acute and 2 of 48 (4%) chronic (p < 0.02). In type B cases, neurologic complications were 15 of 114 (13%) overall; 7 of 22 (32%) were acute cases and 8 of 92 (9%) were chronic (p < 0.004). Early deaths for type B were 12 of 114 (11%) overall; 3 of 22 (14%) acute and 9 of 92 (10%) chronic (p < 0.6). Preoperative hypotension was significant in acute type A patients, with strokes in 2 of 7 (29%) hypotensives compared with 2 of 37 (5%) normotensives (p < 0.05) and early death in 4 of 7 (57%) hypotensives versus 5 of 37 (14%) normotensives (p < 0.009). CONCLUSIONS: Morbidity and mortality for repair of chronic dissection types A and B were acceptable. Preoperative hypotension in acute type A dissection was a major predisposing factor toward stroke (29% versus 5%, p < 0.05). Acute type B dissection had acceptable mortality (14%) but a high rate of neurologic complications (32%). PMID- 9725377 TI - Aortic allograft implantation techniques: pathomorphology and regurgitant jet patterns by Doppler echocardiographic studies. AB - BACKGROUND: The diagnosis of allograft-specific pathology by echocardiography has important consequences for patient counseling and research. This study describes the pathomorphologic findings and color Doppler jet patterns in a consecutive series of patients after allograft placement with either the subcoronary implantation or root replacement technique. METHODS: From 1987 to July 1996, the subcoronary allograft implantation technique and root replacement technique were used in 82 patients and 70 patients, respectively. These patients comprised the study group. RESULTS: The incidence of paravalvular leaks and eccentric regurgitant jets was higher with subcoronary implantation (41%) than with root replacement (11%). Patients with a subcoronary implanted allograft had a higher incidence of eccentric jets. CONCLUSIONS: These findings support the concept of preservation of valve geometry after root replacement, as allograft-specific pathomorphologic abnormalities and eccentric jets are more common after subcoronary implantation of allografts. Learning effects, however, cannot be excluded as the cause of these abnormalities. PMID- 9725378 TI - Preconditioning with cromakalim improves long-term myocardial preservation for heart transplantation. AB - BACKGROUND: Myocardial preservation for heart transplantation relies on hyperkalemic cardiac arrest and hypothermic storage. Our study investigated whether pretreatment with a potassium-channel opener (cromakalim) before prolonged storage in an extracellular fluid improves left ventricular recovery. METHODS: Rabbit hearts were submitted to 6-hours' cold storage and assessed on a blood-perfused isolated heart preparation. Hemodynamic recovery, enzyme release (creatine kinase and lactate dehydrogenase), and adenine nucleotide content were determined. Five groups were tested: control (n=6), no ischemia; UW group (n=7), hearts arrested with and stored in University of Wisconsin solution; STH group (n=5), hearts arrested with and stored in St. Thomas' Hospital solution; cromakalim group (n=6), hearts pretreated with cromakalim (30 microg/kg) before arrest with and storage in St. Thomas' Hospital solution; and glibenclamide group (n=5), hearts pretreated with cromakalim followed by glibenclamide (a potassium channel blocker) before arrest with and storage in St. Thomas' Hospital solution. RESULTS: Hemodynamic recovery was improved and enzyme release was lower in the UW group than in the STH group. Compared with the STH group, the group pretreated with cromakalim had significantly decreased left ventricular end-diastolic pressures, increased left ventricular developed pressures, increased maximal values of positive and negative rates of rise of left ventricular pressure, and increased time constant of isovolumetric relaxation. Hemodynamic recovery was similar in the UW group and cromakalim groups. Glibenclamide did not abolish the effects of cromakalim. None of the protocols affected myocardial energy stores. CONCLUSION: Pretreatment with cromakalim affords additional protection to that provided by cardioplegic arrest and prolonged cold storage using an extracellular solution. The intracellular mechanisms involved remain to be determined. PMID- 9725379 TI - Aortic valve replacement with the Biocor PSB stentless xenograft. AB - BACKGROUND: The midterm clinical results after aortic valve replacement with the Biocor PSB stentless xenograft on all patients operated between October 1992 and October 1996 were reviewed. METHODS: One hundred six patients, aged 70+/-6 years, had aortic valve replacement for aortic stenosis (67%), regurgitation (11%), or both (22%). Associated procedures were done in 49 patients (46%), including coronary artery bypass in 30 patients, mitral valve repair/replacement in 16, and ascending aorta replacement in 5 patients. Aortic cross-clamp and cardiopulmonary bypass times were 96+/-24 and 129+/-31 minutes, respectively. RESULTS: There were 3 (3%) early deaths due to low output (2 patients) and cerebrovascular accident (1 patient). Follow-up of survivors ranged from 6 to 66 months (mean, 39+/-14 months). Survival was 94%+/-2% and 90%+/-3% at 1 and 5 years. There were 5 late deaths due to cardiac cause (2), cancer (2), and pulmonary embolism (1 patient). No patient had structural valve deterioration, whereas 100% and 95%+/-3% were free from valve-related events at 1 and 5 years. There were two reoperations due to narrowing of the left coronary ostium and endocarditis, with an actuarial freedom from reoperation of 99%+/-1% and 98+/-1% at 1 and 5 years, respectively. Functional results demonstrated a mean peak transprosthetic gradient of 16+/-12 mm Hg, with only 1 patient (1%) with a 55 mm Hg gradient. No cases of valve regurgitation greater than mild were recorded at follow-up. Assessment of New York Heart Association functional class demonstrated a significant improvement (2.9+/-0.6 versus 1.4+/-0.7; p=0.01). All patients were free from anticoagulation. CONCLUSIONS: Aortic valve replacement using the Biocor PSB stentless xenograft offers excellent midterm survival, negligible valve deterioration, and a very low rate of valve-related events, which are comparable to estimates reported with other models of stentless xenografts and currently available stented xenografts. Hemodynamic performance is favorable and quality of life satisfactory. PMID- 9725380 TI - Minimal-access aortic and valvular operations, including the "J/j" incision. AB - BACKGROUND: We compared five current minimal-access approaches, namely, parasternal incision, transverse sternotomy, manubrial inverted "T" incision, incomplete mediastinotomy, and our "J/j" incision, to operations in matched patients, including aortic operations. METHODS: In a case-control study of 74 patients, 37 individuals consecutively underwent minimal-access operations (aortic valve, 18, including one mitral valve operation; composite valve graft, six, including one arch and one transaortic mitral valve operation for a patient with Marfan's syndrome; ascending aorta operation, two; root repair/reconstruction, three; mitral valve repair/replacement, seven, including one maze operation; and atrioseptal defect repair, one). The patients were matched by sex, age, surgeon, and operation with 37 control patients who had standard incisions. Patients having the "J/j" incision (n=25) had sternotomies from the first right intercostal space, or sternal notch, to the third to fifth right intercostal space. RESULTS: Minimal-access patients had a shorter postoperative hospital stay than standard incision patients (6.2 versus 8.2 days; p=0.0055), and required similar volumes of blood (0.86 versus 1.03 units; p=0.7243), postoperative morphine dosages (28 mg versus 40 mg, p=0.0643), and oral narcotics (8.1 versus 10.0 doses; p=0.3562). "J/j" incision patients, however, required less morphine (20.6 mg versus 40.9 mg; p=0.0028), but not fewer doses of oral narcotics (7.5 versus 9.9 doses; p=0.2640) and had the shortest postoperative stay (5.1 versus 8.1 days; p < 0.0001). No stroke or clinically noted neurocognitive deficit developed. One minimal-access patient (1/37, 2.7%) with severe preoperative pulmonary morbidity died of adult respiratory distress syndrome. Sternal nonunion developed in 1 patient with an inverted "T" manubrial incision. In a further seven patients, the "J/j" incision was used without a problem, for a total of 32 patients. This compared with a consecutive series of 125 aortic valve replacement operations without a death and 181 patients undergoing ascending arch operations with two 30-day hospital deaths (1.1%) and two strokes (1.1%). CONCLUSIONS: Minimal-access incisions are associated with shorter hospital stays. For the "J/j" incision, even if used for more extensive double-valve, ascending aortic arch, or composite valve operations, postoperative pain appears to be less and patients are discharged even earlier. PMID- 9725382 TI - Up to eight years' follow-up of 997 patients receiving the CarboMedics prosthetic heart valve. AB - BACKGROUND: The aim of the study was to evaluate our clinical experience with the CarboMedics Heart Valve Prosthesis. METHODS: Nine hundred ninety-seven consecutive patients underwent mechanical valve implantation (aortic, 771; mitral, 169; double, 52; tricuspid, 5) with this prosthesis from September 1987 through December 1993. The mean age was 62.3+/-13.7 years (range, 0.4 to 84 years); 56.6% (564 patients) were men. Four hundred seventy patients (47.1%) underwent additional surgical procedures. Mean follow-up was 4.1+/-2.2 years (range, 0 to 8.3 years) with a total of 4,040 patient-years. RESULTS: Early mortality was 5.0% (50/997; aortic, 4.4%; mitral, 6.4%; double, 9.6%). Late mortality was 14.8% (140/947). Survival at 7 years was 75.9%+/-1.8% (aortic, 78.4%+/-2%; mitral, 70.7%+/-4.5%; double, 60.8%+/-7.4%). When matched for sex and age and compared with the normal Norwegian population, our patients had an increased standard mortality ratio in both men (1.9+/-0.4) and women (2.9+/-0.6). The linearized rate of major thromboembolism was 0.9% per patient-year, valve thrombosis 0.2% per patient-year, major bleeding event 0.6% per patient-year, paravalvular leak needing reoperation 0.5% per patient-year, prosthetic valve endocarditis 0.1% per patient-year, and of all reoperations 0.6% per patient year. CONCLUSIONS: The CarboMedics Heart Valve Prosthesis has incidences of morbid events comparable with or better than reported for other mechanical valves. PMID- 9725381 TI - Na+/H+ exchange inhibition improves long-term myocardial preservation. AB - BACKGROUND: Na+/H+ exchange plays an important role in the ionic changes observed during myocardial ischemia and reperfusion. We investigated the cardioprotective efficacy of a selective Na+/H+ exchange inhibitor, 4-isopropyl-3-methylsulfonyl benzoylguanidin-methanesulfonate (HOE642), in a canine model of long-term heart preservation. METHODS: Canine donor hearts were stored for 24 hours in hyperkalemic crystalloid cardioplegic solution; in cardioplegic solution enriched with HOE642; in cardioplegic solution enriched with HOE642, with donor and recipient treated with HOE642; in standard cardioplegic solution, with donor and recipient treated with HOE642; or in standard cardioplegic solution, with only the recipient treated. After orthotopic transplantation, pressure-volume relationships were obtained and dogs were weaned from bypass. Morphology was studied. RESULTS: Myocardial compliance was well preserved when donor and recipient were treated. These groups had the lowest myocardial water content, and no morphologic signs of irreversible damage. In these groups, weaning from cardiopulmonary bypass was successful in 10 of 10 animals, with a cardiac index around 2 L x min(-1) x m(-2). Only 3 of 5 animals in each of the other three groups could be weaned, with significantly lower cardiac indices. CONCLUSIONS: Treatment with HOE642 in both donor and recipient improves myocardial compliance, postweaning cardiac index, and ultrastructure of donor hearts preserved for 24 hours and orthotopically transplanted. PMID- 9725383 TI - Inhibitory effect of photooxidation on intimal and medial thickening of saphenous vein. AB - BACKGROUND: The inhibitory effect of short-term photooxidation on medial and neointimal proliferation of human saphenous vein was investigated. METHODS: Culture medium-filled surgically prepared saphenous vein segments were photooxidized in 0.01% methylene blue solution for 5 minutes. Photooxidized and nonphotooxidized saphenous veins were checked for viability of endothelial cells by culturing vein segments for 21 days followed by histologic and immunohistochemical studies. RESULTS: Endothelial cells of saphenous vein segments remained unaffected after photooxidation. Both the intima and media of nonphotooxidized veins became highly cellular and thickened because of the proliferation and migration of smooth muscle cells. Like precultured fresh saphenous vein, intimal (0.031+/-0.017 mm; p=0.0067) and medial thicknesses (0.702+/-0.123 mm; p < 0.0001) and proliferating cell nuclear antigen-positive cell count (14+/-8/mm2; p=0.0005) of cultured photooxidized veins were significantly less than those of cultured nonphotooxidized veins (intimal thickness, 0.059+/-0.041 mm; medial thickness, 0.997+/-0.228 mm; proliferating cell nuclear antigen positive cell count, 34+/-16/mm2. CONCLUSIONS: Methylene blue-induced short-term photooxidation is effective in inhibition of intimal and medial thickening of saphenous vein. PMID- 9725384 TI - Mechanical properties of human saphenous veins from normotensive and hypertensive patients. AB - BACKGROUND: Different reactivities of saphenous vein grafts in hypertensive and normotensive patients could lead to differences in the postoperative patency of the grafts. METHODS: In saphenous vein rings isolated from remnants of aorta coronary grafts obtained from hypertensive and normotensive patients we studied the length-tension relationship; response to high levels of potassium, norepinephrine, and epinephrine; and relaxation in response to calcium deprivation. RESULTS: The rings from hypertensive patients were stiffer and developed more force (grams force/grams weight) than the rings from normotensive subjects to 80 mmol/L potassium (59+/-16 versus 25+/-5, p < 0.05) and to 1 micromol/L norepinephrine (61+/-8 versus 36+/-7, p < 0.05), but not to 10 micromol/L epinephrine (57+/-11 and 54+/-11; not significant). The rings from hypertensive patients relaxed more slowly than those of the normotensive subjects in a calcium-free medium (time to half-relaxation of 976+/-180 versus 548+/-81 seconds; p < 0.05). CONCLUSIONS: The saphenous vein from hypertensive patients is less distensible, slower to relax, and more reactive to at least two agonists. These differences could influence the graft's patency and the clinical outcome. PMID- 9725385 TI - Cold blood cardioplegia or intermittent cross-clamping in coronary artery bypass grafting? AB - BACKGROUND: We determined that cold blood cardioplegia and intermittent ventricular fibrillation with ischemia were associated with similar enzyme and myocardial protein leakage in a randomized, prospective study of 40 patients. We have continued to use both methods in our unit, according to surgeons' preference. METHODS: In our database we have reviewed 1,923 patients who have undergone first-time elective or urgent coronary artery bypass grafting from January 1992 to May 1997. RESULTS: Five hundred seventy-eight patients underwent coronary artery bypass grafting with cold blood cardioplegia and 1,345 had ventricular fibrillation and aortic cross-clamping. The preoperative factors were virtually identical. Intraoperative differences were only those inherent to the two techniques: temperature and cross-clamp time. Mortality was 2.5% for ventricular fibrillation and aortic cross-clamping arrest and 2.1% for cardioplegia (p=0.55 by chi2 test). There was a higher use of the intraaortic balloon pump in the ventricular fibrillation and aortic cross-clamping group (2.4% versus 1.0%; p=0.04), but no other differences in outcome were detected. CONCLUSIONS: A truly randomized trial to demonstrate which technique is superior is impractical at this level of difference because it would require 37,000 patients to avoid a beta error. We have to base our practice on the retrospective data available. Each technique has its merits in practice, which are discussed. PMID- 9725386 TI - Effects of cardioplegic solutions on the vasoreactivity of the internal mammary artery. AB - BACKGROUND: During free internal mammary artery grafting, cardioplegia administration can be performed through the internal mammary artery. The present study examined whether cardioplegic solutions produce arterial graft constriction and functional endothelial damage. METHODS: Forty internal mammary artery segments from 10 patients were incubated in Krebs solution (n=10), University of Wisconsin solution (n=10), Broussais Hospital solution (n=10), or blood cardioplegia (n=10). RESULTS: There was a significant difference in sensitivity to norepinephrine between segments in Krebs solution and those in University of Wisconsin solution or Broussais Hospital solution but not segments in blood cardioplegia. There was a significant difference in relaxation to acetylcholine between segments in Krebs solution and those in the three other cardioplegic solutions and between those in blood cardioplegia and segments in University of Wisconsin solution or Broussais Hospital solution. There was no significant difference in relaxation to sodium nitroprusside between segments in any of the solutions. CONCLUSIONS: These experiments suggest that storage in the different cardioplegic solutions studied does not preserve the initial vasoreactivity of the internal mammary artery. However, blood cardioplegia appears to be less deleterious in regard to endothelial and myogenic vascular function. PMID- 9725387 TI - Temporary luminal arteriotomy seal: II. Coronary artery bypass grafting on the beating heart. AB - BACKGROUND: This study assessed the feasibility of applying a temporary luminal arteriotomy seal during end-to-side coronary artery bypass grafting on the beating heart. METHODS: In 18 consecutive pigs, the left internal mammary artery was grafted to the left anterior descending coronary artery, and the arteriotomy was temporarily sealed luminally by a 200-microm-thick polyurethane seal. Endothelial denudation, medial necrosis, and intimal hyperplasia were measured quantitatively and compared with conventionally sutured anastomoses (n=4 pigs). RESULTS: Insertion and retrieval of the seal required 28+/-12 and 11+/-6 seconds, respectively. Including the arteriotomy, coronary artery occlusion was limited to about 80 seconds. The seal provided a bloodless arteriotomy in all anastomoses with unimpeded coronary artery blood flow. Endothelial denudation was limited to two thirds of the circumference of the coronary artery. No medial necrosis was found. Intimal hyperplasia at the suture line was small, although more pronounced when compared with conventionally sutured anastomoses. CONCLUSIONS: In off-pump, beating-heart coronary artery bypass grafting, the temporary luminal arteriotomy seal provided a bloodless arteriotomy with negligible obstruction to coronary artery blood flow, and with a minimum of arterial wall damage. It is conceivable that this seal may expand the indications for coronary surgical procedures without cardiopulmonary bypass. PMID- 9725388 TI - Coronary bypass flow during use of intraaortic balloon pumping and left ventricular assist device. AB - BACKGROUND: Intraaortic balloon pumping (IABP) and left ventricular assist device (LVAD) are used for left ventricular support when low cardiac output occurs after a coronary bypass operation for serious coronary artery disease. There are hemodynamic differences in blood flow in various kinds of coronary artery bypass grafts, caused by their inherent physiologic characteristics. The hemodynamic effects of left ventricular assistance with IABP and LVAD on blood flow through various coronary artery bypass grafts were investigated. METHODS: An ascending aorta-coronary bypass graft (ACB), an internal thoracic artery, and a descending aorta-coronary bypass graft were anastomosed to the left anterior descending coronary artery in a canine model. In this experimental model, the blood flow to the same coronary bed in the three types of grafts could be evaluated. Blood flow in the left anterior descending coronary artery through the three types of coronary bypass grafts was studied in this model during or in the absence of ventricular assistance. RESULTS: In the control study, the systolic blood flow did not differ among the three types of grafts, but the diastolic flow decreased in the following order: with the ACB, the internal thoracic artery, and the descending aorta-coronary bypass graft. The systolic flow during IABP and LVAD was similar to the control flows. Use of IABP increased the diastolic flow by 75.3%+/-12.4% of the control value in the ACB, 37.9%+/-25.0% in the internal thoracic artery, and 21.2%+/-11.4% in the descending aorta-coronary bypass graft. The LVAD increased the diastolic flow by 97.7%+/-18.7% of the control value in the ACB, 64.5%+/-25.7% in the internal thoracic artery, and 63.0%+/-27.9% in the descending aorta-coronary bypass graft. The diastolic blood flows in the left anterior descending coronary artery and the three types of grafts were significantly greater with IABP than the control values, and significantly greater with LVAD than with IABP and the control values. The degrees of increase of diastolic flows in the left anterior descending coronary artery and the ACB with IABP and LVAD were significantly greater than in the arterial grafts (p < 0.01). CONCLUSIONS: The diastolic flows in the internal thoracic artery and descending aorta-coronary bypass graft increased less than in the native left anterior descending coronary artery and ACB during left ventricular assistance, particularly with IABP. It is important for the selection of tactics for the management of catastrophic status after coronary bypass grafting to consider the hemodynamic characteristics of the graft. PMID- 9725389 TI - Vivostat system autologous fibrin sealant: preliminary study in elective coronary bypass grafting. AB - BACKGROUND: The Vivostat System is a medical device for the preparation of an autologous fibrin sealant from 120 mL of the patient's blood in the operating room. The system is fully automated and microprocessor controlled and is made up of three components: an automated processor unit, an automated applicator unit, and a disposable, single-patient-use unit, which includes a preparation set and a Spraypen applicator. The biochemical process is initiated by batroxobin, which acts upon the fibrinogen in the patient's plasma. The completion of the process depends entirely on endogenous thrombin in producing the sealant. METHODS: Twenty four volunteer patients undergoing elective primary coronary artery bypass grafting were randomized to either conventional hemostasis (control group) or the use of Vivostat fibrin sealant as an adjunct to conventional hemostasis. The patients were followed up at 1 month and 1 year. RESULTS: The preparation process was completed in 30 minutes. No safety issues associated with the use of the sealant were identified. From 120 mL of the patient's blood the yield of fibrin sealant was 4.5 mL (range, 3.9 to 4.8 mL). There was a favorable trend toward lower amounts of chest tube drainage in the Vivostat group. In the Vivostat group, 1 of 11 patients (9%) required a perioperative transfusion and in the control group 3 of 12 patients (25%) required a perioperative transfusion. CONCLUSIONS: It is possible to prepare autologous fibrin sealant with the Vivostat system in 30 minutes. No exogenous thrombin is required. The sealant has no known adverse effects and may prove to be a useful adjunct to hemostasis in cardiothoracic surgery. PMID- 9725390 TI - Beneficial effects of angiotensin-converting enzyme inhibitors during acute revascularization. AB - BACKGROUND: This experimental study was undertaken to determine whether using angiotensin-converting enzyme inhibitors during surgical revascularization of acutely ischemic myocardium would improve wall motion and limit infarct size. METHODS: Twenty pigs underwent 90 minutes of occlusion of the second and third diagonal arteries followed by 45 minutes of cardioplegic arrest and 180 minutes of reperfusion. In 10 animals, the angiotensin-converting enzyme inhibitor enalaprilat (0.05 mg/kg) was infused intravenously during coronary occlusion; 10 other animals received no angiotensin-converting enzyme inhibitors. Ischemic damage was assessed by the number of cardioversions required for ventricular tachycardia or fibrillation; wall motion scores using echocardiography (4=normal to -1=dyskinesia); and infarct size using histochemical staining. Epicardial coronary artery vasomotor function was assessed using standard organ chamber methodology. RESULTS: Enalaprilat-treated hearts had the least amount of ventricular irritability (0.84+/-0.24 versus 2.77+/-0.22 cardioversions; p < 0.01), the best recovery of wall motion score (3.20+/-0.15 versus 1.52+/-0.07; p < 0.0001), and the lowest infarct size (22.6%+/-1.4% versus 37.7%+/-3.0%; p < 0.001). Endothelium-independent relaxation was preserved in all hearts; however, endothelium-dependent relaxation was impaired in both groups. CONCLUSIONS: Angiotensin-converting enzyme inhibitors reduce myocardial damage during surgical revascularization of acutely ischemic myocardium. PMID- 9725391 TI - Experience with antegrade bihemispheric cerebral perfusion in aortic arch operations. AB - BACKGROUND: Various techniques have been used for cerebral protection in aortic arch operations. Antegrade cerebral perfusion has lost its popularity to hypothermic circulatory arrest to overcome the so-called cluttered operative field. Hypothermic circulatory arrest has its own problems of coagulopathy, time constraints, and prolongation of cardiopulmonary bypass time. METHODS: Since June 1986 we have used antegrade bihemispheric cerebral perfusion with moderate hypothermia in 20 patients with aortic arch disease. Twelve patients had aneurysm, 7 had dissection, and 1 had traumatic tear. Five patients had had previous sternotomy for ascending aortic replacement. In addition to arch reconstruction, 7 patients had aortic valve replacement or repair, 2 patients had Bentall procedure, and 3 had selective innominate reconstruction. The mean cerebral perfusion time was 51+/-29 minutes. In 7 patients the cerebral perfusion time was between 60 and 120 minutes. RESULTS: There was no in-hospital or 30-day mortality. The blood product requirements were significantly less with moderate hypothermia. One patient suffered cerebrovascular accident (5%). None of the 7 patients with cerebral perfusion times of 60 to 120 minutes had any neurologic deficits. These results are superior to those reported for hypothermic circulatory arrest with or without retrograde cerebral perfusion. CONCLUSIONS: Antegrade bihemispheric cerebral perfusion is an optimal adjunct for cerebral protection during aortic arch operations. PMID- 9725392 TI - Adrenomedullin in patients at high risk for pulmonary hypertension. AB - BACKGROUND: Adrenomedullin is a newly identified peptide with profound hypotensive effects. We investigated perioperative adrenomedullin levels among patients with congenital heart disease with and without pulmonary hypertension. METHODS: Levels of plasma adrenomedullin, endothelin-1, and nitric oxide metabolites were measured in three groups: (1) low pulmonary flow (n=11); (2) high flow/low pulmonary arterial pressure (less than 60% systemic pressure) (n=9); and (3) high flow/high pressure (n=10). Samples were obtained preoperatively, on and off pump, and 3, 6, and 12 hours after bypass. RESULTS: Adrenomedullin levels were highest in the low pulmonary flow group (189.7+/-15 pg/mL low flow versus 103.1+/-9.5 pg/mL high flow/low pulmonary and 139+/-17.5 pg/mL high flow/high pressure at 12 hours; p < or = 0.05). The arterial pressure/systemic pressure remained significantly lower in the high flow/low pulmonary pressure compared with the high flow/high pressure group (0.37+/-0.08 versus 0.62+/-0.11; p < 0.005). Perioperative endothelin-1 and nitric oxide levels remained low in the low pulmonary flow group but increased progressively in both high flow groups. CONCLUSIONS: Circulating plasma adrenomedullin appears to affect baseline vascular tone in patients with intact endothelial function. It may interact with nitric oxide and endothelin-1 to help regulate blood pressure perioperatively in patients with congenital heart disease. PMID- 9725393 TI - A 26-year experience with surgical management of tetralogy of Fallot: risk analysis for mortality or late reintervention. AB - BACKGROUND: Over the past decade repair of tetralogy of Fallot (TOF) in infancy has gained favor. It is still uncertain what effect early complete repair will have on survival or late reoperation on the right ventricular outflow tract. METHODS: To assess these outcomes, we reviewed our experience (1971-1997) with 294 patients undergoing operation at one institution. Median follow-up was 10.6 years (range, 0.1 to 26 years), and was complete for 90.2% patients. RESULTS: Primary complete repair was done in 199 patients (68%), and a staged repair in 62 patients (21%). Thirty-three patients had only a palliative procedure. Sixty eight patients (23.1%) had complex pathologic processes, including pulmonary atresia in 53. Hospital mortality for primary repair was 11.1% (22/199), for staged repair was 17.7% (11/62), and for palliative procedures was 15.5% (16/103 procedures). Since 1990 mortality has been 2.1%, 11.8%, and 0% respectively (p < 0.001), despite younger age at repair (0.6+/-0.1 versus 2.1+/-0.2 years; p < 0.001). Multivariate analysis identified longer period of hypothermic circulatory arrest, pulmonary artery patch angioplasty, earlier year of operation, and closure of the foramen ovale as risk factors for hospital death. For hospital survivors 20-year survival was 98%+/-3% for TOF with pulmonary stenosis and 88%+/ 9% for TOF with pulmonary atresia (p=0.09). Reintervention on the right ventricular outflow tract was needed in 14.1% (37/261) patients. Freedom from reintervention on the right ventricular outflow tract at 20 years was 86%+/-4% for TOF with pulmonary stenosis and 43%+/-16% for TOF with pulmonary atresia (p=0.001). For the subgroup TOF with pulmonary stenosis, this was 85%+/-5% after primary repair and 91%+/-8% after staged repair (not significant). At 15-year follow-up, this was 78%+/-10% for patients not older than 1 year at operation compared with 88%+/-4% for older patients (not significant). CONCLUSIONS: Early mortality after primary repair of TOF has significantly improved and late survival is excellent. Primary repair in infancy does not increase risk for reintervention on the right ventricular outflow tract. PMID- 9725394 TI - Effects of modified and classic Blalock-Taussig shunts on the pulmonary arterial tree. AB - BACKGROUND: The aim of this study was to assess by angiography the late effects of both classic and modified Blalock-Taussig shunts in terms of growth and development of stenosis and distortion. METHODS: At a mean of 51 months postoperatively, we retrospectively reviewed the results in 78 patients who underwent creation of Blalock-Taussig shunts (25 classic and 71 modified). RESULTS: At the level of the anastomosis, the shunt caused a reduction in diameter of the pulmonary artery in 49% of all shunts, major stenosis (>50% narrowing of the lumen) in 14%, and distortion of the pulmonary artery in 19%, findings that did not correlate with the type of shunt. Distortion did correlate with younger age at the time of shunt operation (p=0.01). CONCLUSIONS: After a Blalock-Taussig shunt, growth of the pulmonary arteries occurred but did not exceed the normal growth of the pulmonary arterial tree. Moreover, a shunt procedure can cause distortion and stenosis of the pulmonary artery, which may have important implications for future corrective surgical intervention. All these findings support earlier complete surgical repair of correctable congenital cardiac defects. PMID- 9725395 TI - Neonatal mechanical bridging to total orthotopic heart transplantation. AB - BACKGROUND: Until recently, newborns with medically intractable cardiac failure caused by congenital malformations were mostly doomed to death because of the severity of the disease, which precludes a palliative operation, or because of fatal deterioration before availability of a suitable donor heart. METHODS: The recently developed paracorporeal pneumatically driven Medos HIA ventricular assist device offers a therapeutic option for these small infants because it is manufactured in various sizes and is even suitable for cardiac assistance in neonates with a body surface area less than 0.3 m2. RESULTS: We report our initial experience with this device, which we used for univentricular bridging to total orthotopic cardiac transplantation in 3 infants. The device was inserted to support the left ventricle in two instances and to support the right heart in one. Successful bridging to transplantation was achieved in 2 infants for periods of 2 and 7 weeks. CONCLUSIONS: Our experience demonstrates the feasibility of univentricular mechanical support followed by successful cardiac transplantation in infants and newborns. PMID- 9725396 TI - Total cavopulmonary anastomosis (Fontan) in children with Down's syndrome. AB - BACKGROUND: There is a paucity of information to guide the management of the child with Down's syndrome and congenital heart disease in whom biventricular repair is precluded. METHODS: Through the cardiology and cardiovascular surgery databases of The Hospital for Sick Children and Toronto Congenital Cardiac Centre for Adults, we identified patients with trisomy 21 and ventricular hypoplasia who had undergone a Fontan procedure (or modification). RESULTS: Of 533 patients who had undergone a Fontan operation between 1976 and 1997, 4 had trisomy 21. All 4 patients had unbalanced complete atrioventricular septal defect with right ventricular hypoplasia in 3 and left ventricular hypoplasia in 1. Three patients survived, and 1 died of endocarditis. The 3 survivors have done well in the short term and medium term without complications related to the pulmonary vasculature. CONCLUSIONS: We suggest that in appropriately selected patients with trisomy 21 and ventricular hypoplasia who are unsuitable for two or one and a half ventricle repair, the Fontan procedure is not contraindicated and provides short-term and medium-term benefit. PMID- 9725397 TI - Upregulated and downregulated transcription of myocardial genes after pulmonary artery banding in pigs. AB - BACKGROUND: Acute or chronic pressure overload may occur during or after cardiac surgical procedures. Typical examples are aortic cross-clamping and pulmonary artery banding. It is well known that mechanical stress induces transcription of different myocardial genes. However, these results were mainly obtained from in vitro studies and experiments with rodents. This experiment was carried out to investigate molecular alterations after pressure overload in porcine hearts. METHODS: The study was performed with 35 Landrace pigs with a mean weight of 32+/ 1.2 kg. The five groups consisted of 7 pigs each, 3 sham-operated pigs and 4 banded pigs. The hearts were exised after different time intervals. We investigated messenger RNA expression of sarcoplasmic reticulum adenosine triphosphatase, phospholamban, alpha-/beta-myosin heavy chain, and atrial natriuretic factor by Northern blot analysis. RESULTS: The ratio of right ventricular weight to body weight increased significantly after 7 and 24 days in banded pigs (p < 0.05). Atrial natriuretic factor messenger RNA was significantly upregulated in banded pigs versus sham-operated pigs after 1 day (240%+/-7% versus 100%+/-6%; p < 0.01) and 3 days (520%+/-8% versus 100%+/-8%; p < 0.01). There was insignificant downregulation of sarcoplasmic reticulum adenosine triphosphatase and phospholamban after 1, 3, and 7 days. Myosin heavy chain messenger RNA expression remained unchanged. CONCLUSIONS: Pulmonary artery banding results in hypertrophic response of the porcine right ventricle; however, the weight increase is not the result of myosin heavy chain messenger RNA upregulation. Atrial natriuretic factor messenger RNA is locally expressed in mechanically stressed myocytes. Furthermore, pressure overload downregulates transcription of calcium-binding proteins that can influence ventricular contractility. These results may have an impact on cardiac surgical procedures. PMID- 9725398 TI - Decreased exhaled nitric oxide may be a marker of cardiopulmonary bypass-induced injury. AB - BACKGROUND: Nitric oxide is an endothelium-derived vasodilator. Cardiopulmonary bypass may induce transient pulmonary endothelial dysfunction with decreased nitric oxide release that contributes to postoperative pulmonary hypertension and lung injury. Exhaled nitric oxide levels may reflect, in part, endogenous production from the pulmonary vascular endothelium. METHODS: We measured exhaled nitric oxide levels before and 30 minutes after cardiopulmonary bypass in 30 children with acyanotic congenital heart disease and left-to-right intracardiac shunts undergoing repair. RESULTS: Exhaled nitric oxide levels decreased by 27.6%+/-5.6% from 7+/-0.8 to 4.4+/-0.5 ppb (p < 0.05) 30 minutes after cardiopulmonary bypass despite a reduction in hemoglobin concentration. CONCLUSIONS: The decrease in exhaled nitric oxide levels suggests reduced nitric oxide synthesis as a result of pulmonary vascular endothelial or lung epithelial injury. This may explain the efficacy of inhaled nitric oxide in the treatment of postoperative pulmonary hypertension. Furthermore, strategies aimed at minimizing endothelial dysfunction and augmenting nitric oxide production during cardiopulmonary bypass may decrease the incidence of postoperative pulmonary hypertension. Exhaled nitric oxide levels may be useful to monitor both cardiopulmonary bypass-induced endothelial injury and the effect of strategies aimed at minimizing such injury. PMID- 9725399 TI - Hematologic and economic impact of aprotinin in reoperative pediatric cardiac operations. AB - BACKGROUND: Aprotinin consistently reduces blood loss and transfusion requirements in adults during and after cardiac surgical procedures, but its effectiveness in children is debated. We evaluated the hemostatic and economic effects of aprotinin in children undergoing reoperative cardiac procedures with cardiopulmonary bypass. METHODS: Control, low-dose aprotinin, and high-dose aprotinin groups were established with 15 children per group. Platelet counts, fibrinogen levels, and thromboelastographic values at baseline and after protamine sulfate administration, number of blood product transfusions, and 6 hour and 24-hour chest tube drainage were used to evaluate the effects of aprotinin on postbypass coagulopathies. Time needed for skin closure after protamine administration and lengths of stay in the intensive care unit and the hospital were recorded prospectively to determine the economic impact of aprotinin. RESULTS: Coagulation tests performed after protamine administration rarely demonstrated fibrinolysis but did show significant decreases in platelet and fibrinogen levels and function. The thromboelastographic variables indicated a preservation of platelet function by aprotinin. Decreased blood product transfusions, shortened skin closure times, and shortened durations of intensive care unit and hospital stays were found in the aprotinin groups, most significantly in the high-dose group with a subsequent average reduction of nearly $3,000 in patient charges. CONCLUSIONS: In children undergoing reoperative cardiac surgical procedures, aprotinin is effective in attenuating postbypass coagulopathies, decreasing blood product exposure, improving clinical outcome, and reducing patient charges. PMID- 9725400 TI - Pulmonary function after one-lung ventilation in newborns: the basis for neonatal thoracoscopy. AB - BACKGROUND: To maintain good exposure during major video-assisted thoracic surgery it is necessary to deflate completely the ipsilateral lung. However, little is known about the effects of one-lung ventilation (OLV) on pulmonary function in newborn patients. METHODS: Ten neonatal domestic pigs with a mean age of 6+/-0.6 days were intubated and ventilated in pressure-controlled mode (inspired oxygen fraction=1.0). One-lung ventilation was maintained for 120 minutes. Serial measurements of hemodynamics and gas exchange were done before, during, and until 90 minutes after OLV. Pulmonary function testing was performed before and after OLV for each lung separately. RESULTS: With the inspired oxygen fraction set at 1.0, arterial oxygen saturation remained stable at 100% during OLV. Venous admixture and alveolar-arterial oxygen tension gradient increased slightly from the baseline value of 2.6% +/-0.3% to 3.8%+/-0.3% during OLV (mean+/-standard error of the mean; p=0.02), and from 358+/-28 to 407+/-18 mm Hg (not significant), respectively. Both values returned to baseline during the subsequent ventilation of both lungs. Static compliance and resistance of the ventilated lung did not change. Compliance of the collapsed lung decreased after reexpansion from 0.42+/-0.07 to 0.29+/-0.06 mL x cm H2O(-1) x kg(-1), p=0.008). Resistance remained unchanged (0.22+/-0.02 versus 0.25+/-0.05 cm H2O x L(-1) x s( 1); not significant). CONCLUSIONS: There were only minor effects on pulmonary function during and after OLV in the neonatal piglet. Alterations in gas exchange during OLV were minimal. Prolonged collapse of the lung with subsequent reexpansion was associated with a slight decrease in compliance, indicating some mild lung injury. PMID- 9725401 TI - Mediastinal germ cell tumor in a child with precocious puberty and Klinefelter syndrome. AB - An 8-year-old boy presented with precocious puberty and a mediastinal mass. A computer search showed that this rare presentation is most common with germ cell tumor of the mediastinum in children with Klinefelter syndrome. The tumor was completely resected after preoperative chemotherapy, and the patient is well 2 years after the operation. In patients with Klinefelter syndrome, germ cell tumors are 50 times more common than in patients without Klinefelter syndrome, usually contain nonseminomatous elements, present at an earlier age, and are seldom testicular in location. PMID- 9725403 TI - Delayed presentation of foreign body reaction secondary to retained pacing wires. AB - Temporary pacing wires are often left behind, assumed not to cause problems. We present 2 cases of delayed presentation of anterior mediastinal foreign body reaction secondary to retained pacing wires after coronary operations performed more than 5 years previously. PMID- 9725402 TI - Alternative to reconstruction of the pulmonary outflow tract in the Ross procedure. AB - We report our experience with 2 cases in which we used the native ascending aorta and a porcine valve to reconstruct the right ventricular outflow tract in the Ross procedure. Unfortunately, in many parts of the world, the lack of homografts for reconstruction of the right ventricular outflow tract limits the use of the Ross procedure. The technique described herein can be an alternative to a cryopreserved pulmonary homograft replacement for adult patients. PMID- 9725405 TI - Successful removal of massive cardiac neurilemoma with cardiopulmonary bypass. AB - A 46-year-old woman was referred to our hospital because of cardiac enlargement seen on a chest radiograph. Imaging studies showed a massive intrapericardial tumor with a size of 12x8x7 cm. Tumor dissection included inspection of the inner aspect of the superior vena cava with use of cardiopulmonary bypass, because the mass was tightly adherent to both superior vena cava and right atrium. The pathologic diagnosis was neurilemoma. PMID- 9725404 TI - Thoracic splenosis. AB - Thoracic splenosis is a rare pathologic entity resulting from seeding of splenic tissue in the pleural cavity after thoracoabdominal trauma. A 45-year-old man with a history of splenectomy secondary to abdominal trauma presented with a left lung mass and an inconclusive tissue diagnosis after needle biopsy. Thoracic splenosis was not suspected preoperatively, considered on an intraoperative frozen section, and established on permanent pathologic biopsy specimens obtained during thoracotomy. A history of thoracoabdominal trauma, combined with radiologic and radionuclide imaging studies, may establish the diagnosis without thoracotomy. PMID- 9725406 TI - Recovery from end-stage ischemic cardiomyopathy during long-term LVAD support. AB - A patient with ischemic cardiomyopathy and extremely reduced left ventricular function (left ventricular ejection fraction=0.10) presented to our institution for cardiac transplantation. Because of his worsening condition he was placed on the Novacor left ventricular assist device. During 3 months of support his left ventricular function recovered and he successfully underwent percutaneous transluminal coronary angioplasty and minimally invasive direct coronary artery bypass grafting procedures; subsequently he could be weaned from the left ventricular assist device and discharged. The patient is no longer considered for cardiac transplantation. PMID- 9725407 TI - Intraabdominal hemorrhage during combined coronary artery bypass and carotid endarterectomy. AB - Intraabdominal complications during cardiopulmonary bypass are extremely rare, with an incidence of less than 1% in multiple retrospective studies. These complications are associated with a high mortality, and their rapid diagnosis is critical to the outcome of the patient. We present a case of spontaneous intraabdominal hemorrhage after combined carotid endarterectomy and four-vessel coronary artery bypass grafting, which was diagnosed through a diaphragmatic window. PMID- 9725408 TI - Fetal cardiac tamponade due to an intrapericardial teratoma. AB - A case of an intrapericardial tumor diagnosed in utero at 26 weeks of gestation is presented. The prenatal echocardiographic follow-up of an incipient hydrops fetalis determined the management and the emergency surgical treatment. Histologically, the tumor appeared to be a benign teratoma, grade I. In the postoperative period an unexpected mediastinal tumor was found and removed later. This tumor also appeared to be a benign teratoma, grade 0. Both teratomas were independent and therefore primary. PMID- 9725409 TI - Isolated tricuspid regurgitation caused by a dilated tricuspid annulus. AB - In most of the previously reported cases of isolated tricuspid regurgitation, both tricuspid leaflets and subvalvar tissue have been absent, hypoplastic, or fused. For this reason, tricuspid valvoplasty was difficult and valve replacement was adopted in many cases. In the present case of a 52-year-old man, however, the tricuspid valve showed no abnormalities other than a severely dilated tricuspid annulus. Ring annuloplasty was performed, and this resulted in a subsequent satisfactory course without anticoagulant therapy. PMID- 9725410 TI - Paradoxical embolism of a shotgun pellet. AB - Paradoxical embolism of a projectile from the venous to arterial system is a rare occurrence, which can cause diagnostic confusion. We present a case of venous embolism of a shotgun pellet from the left upper extremity to the noncoronary sinus of the aortic valve across a secundum-type atrial septal defect. Prevention of distal embolism of the pellet was presumably a result of its containment by flow vortices created within the sinuses of Valsalva. PMID- 9725411 TI - Combined therapies for composite graft infection after Bentall's procedure. AB - We present a patient who suffered from composite graft infection and mediastinitis. After replacement of the infected composite graft, in addition to administration of antibiotics, continuous irrigation of the mediastinum with solutions containing povidone-iodine and cefazolin sodium and transposition of the greater omentum were performed. His postoperative course was uneventful. Combined therapies including mediastinal irrigation and omental transposition should be considered after an operation for composite graft infection complicated with mediastinitis. PMID- 9725412 TI - Infection with Mycobacterium tuberculosis complicating a pulmonary sequestration. AB - Pulmonary sequestration is a relatively rare malformation. Infection with common pyogenes is a frequent feature in the evolution of this disease. We report a case of intralobar sequestration infected with Mycobacterium tuberculosis in the absence of any other site of tuberculous infection. The patient underwent surgical removal of the affected lobe and subsequent antituberculous chemotherapy. At 1-year follow-up his clinical status is excellent. PMID- 9725413 TI - Cardiopulmonary bypass with danaparoid sodium and ancrod in heparin-induced thrombocytopenia. AB - Heparin is the standard anticoagulant for patients undergoing cardiopulmonary bypass. There are some patients for whom heparin is unsuitable and ancrod (a defibrinogenating enzyme) has been used as an alternative. We present a patient with heparin-induced thrombocytopenia in whom treatment ancrod was ineffective. The addition of danaparoid sodium (a heparinoid) allowed safe cardiopulmonary bypass. We discuss the reasons for this and suggest that the combination of ancrod and danaparoid sodium is a logical one in such cases. PMID- 9725415 TI - Management of recurrent ventricular tachycardia with ventricular assist device placement. AB - Life-threatening, recurrent ventricular tachycardia developed in a 54-year-old heart transplant candidate with ischemic cardiomyopathy. The episodes of ventricular tachycardia were refractory to aggressive medical management and implantable cardiac defibrillator placement. A Heartmate left ventricular assist device was implanted, in combination with isolated right coronary artery bypass grafting, which abolished any further episode of ventricular tachycardia. The patient successfully underwent cardiac transplantation 79 days later. PMID- 9725414 TI - Hemothorax in type I neurofibromatosis. AB - We report a case of life-threatening hemothorax caused by rupture of a left thyrocervical trunk aneurysm and arteriovenous fistula in a patient with type I neurofibromatosis. This lesion was treated with endovascular coil embolization. PMID- 9725416 TI - Transesophageal echocardiographic evaluation of perioperative coronary sinus trauma. AB - Retrograde cardioplegia is relatively safe, with a rate of coronary sinus rupture of 0.6%. With the advent of perioperative transesophageal echocardiography, it is now possible to detect and evaluate the extent of damage consequent to the use of retrograde cardioplegia and better formulate an intraoperative course of action. The evolution of these lesions can also be monitored by transesophageal echocardiography during the postoperative period. PMID- 9725417 TI - Off-pump coronary bypass grafting: how to use the Octopus Tissue Stabilizer. AB - Off-pump coronary artery bypass grafting requires immobilization of the coronary artery. A suction device (Octopus Tissue Stabilizer), attached to the epicardium and connected rigidly to the operating table rail, was used through limited and full surgical access for single-vessel and multivessel arterial revascularization, respectively. An outline for its application, as used by us to construct 122 anastomoses in 70 patients, including posterior wall grafting (in 9 patients) and sequential grafting on the anterior wall (in 17 patients), is presented. PMID- 9725418 TI - Training model for "beating-heart" coronary artery anastomoses. AB - The principal technical challenge in "off-pump" coronary surgery is to perform an accurate coronary anastomosis on a beating heart. For the purpose of training our residents in performing off-pump coronary artery anastomoses, we have developed a mechanical heart simulator in which trainees can practice performing these anastomoses repeatedly until a satisfactory level of skill and confidence is attained. PMID- 9725419 TI - Muscle-sparing anterior thoracotomy for one-stage bilateral lung volume reduction operation. AB - Bilateral lung volume reduction produces significant clinical and physiologic improvement in selected patients with end-stage emphysema. Current surgical approaches consist of median sternotomy and video-assisted thoracoscopy. This report describes an alternate technique of single-stage, bilateral lung volume reduction using muscle-sparing anterior thoracotomy in 18 patients with severe lung emphysema. PMID- 9725421 TI - Modified subclavian flap aortoplasty for coarctation repair in patients less than three months of age. AB - Subclavian flap aortoplasty is one of the best procedures for coarctation repair, but recoarctation still remains a problem in neonates and infants. We employed subclavian flap aortoplasty with resection of the whole layer of aortic ductal tissue in 9 patients less than 3 months of age with coarctation of the aorta and obtained satisfactory results. PMID- 9725420 TI - Pericardial closure using fascia lata in patients undergoing pneumonectomy with pericardiectomy. AB - In patients undergoing pneumonectomy with concomitant pericardiectomy, patch closure of the defect in the pericardium is required to prevent postoperative cardiac herniation. We used harvested autologous fascia lata as a pericardial patch in such operations on 6 patients with progressive lung cancer after induction chemoradiotherapy. PMID- 9725422 TI - Axillary thoracoscopy. AB - Thoracoscopy can be done safely and effectively through working ports placed in the axilla in patients whose pathology is in the upper half of the thorax. We have used this technique successfully in 37 patients with no complications. Advantages include superior cosmesis, optimal access to the apex of the chest, and, if necessary, easy conversion to axillary thoracotomy. PMID- 9725423 TI - Minimally invasive management for first and recurrent pneumothorax. AB - Minimally invasive techniques for treatment of pneumothorax should yield the standard of results set with open procedures: the operative morbidity should remain less than 15%, and the recurrence rate less than 1%. In the era before video-assisted thoracic surgery, two minimally invasive variants were used. Chemical pleurodesis resulted in an unsatisfactory recurrence rate of at least 15%. In contrast, pleurectomy and apical stapling performed through a transaxillary minithoracotomy compared favorably with larger thoracotomy approaches, and allowed a reduced hospital stay. Evaluation of video-assisted thoracic surgical operations for spontaneous pneumothorax is hampered by a lack of controlled studies. The general impression is that morbidity did not decline significantly; the main determinant of complications is the patient's underlying health status. However, published recurrence rates range from 5% to 10%, in spite of a shorter follow-up time span. Optimized results are achieved when classic principles combining apical wedge resection and pleurodesis are applied. Reduction of hospital stay is not only a result of the new technology, but also changing drainage and discharge policies. Reduction of cost is debatable, because many studies do not consider the cost of video equipment. The main advantage when compared with open thoracotomy is reduction of postoperative pain. The only two available controlled studies conclude that there is no obvious advantage of video assisted thoracic surgery when compared with conventional limited-access surgery. The future role of video-assisted thoracic surgery in this disease remains to be determined by a large-scale prospective evaluation. PMID- 9725424 TI - If I were a young thoracic surgeon... PMID- 9725425 TI - Coarctation with valvular lesions in adults. PMID- 9725426 TI - Postoperative mediastinitis and beta-adrenergic drugs. PMID- 9725427 TI - Sternal dehiscence in poststernotomy mediastinitis. PMID- 9725428 TI - Ascending versus descending balloon counterpulsation. PMID- 9725429 TI - Aortic coarctation associated with aortic or mitral valve disease: which lesion to correct first? PMID- 9725430 TI - Pneumoperitoneum for prolonged air leaks after lower lobectomies. PMID- 9725431 TI - Recurrence of the mediastinal growing teratoma syndrome. PMID- 9725432 TI - Operation on the thoracic aorta. PMID- 9725433 TI - Percutaneous drainage of pericardial cyst with right-sided heart failure. PMID- 9725434 TI - A simple method of aortic root venting for CABG. PMID- 9725435 TI - Aortic root hemorrhage and presealed composite grafts with porous sewing cuffs. PMID- 9725436 TI - Air leaks and pleural spaces after lung volume reduction. PMID- 9725437 TI - Modern management of traumatic rupture of the aortic isthmus. PMID- 9725438 TI - Left atrial catheter removal. PMID- 9725439 TI - One and one-half ventricle repairs. Introduction. PMID- 9725440 TI - What is a ventricle? AB - BACKGROUND: The concept of "one and a half" ventricular repair is to use "half" a ventricle to support the pulmonary circulation. The component makeup of any ventricle needs clarification for us to understand the nature of the so-called half ventricle. METHODS: The components of normal and abnormal ventricles are reviewed. RESULTS: Normal ventricles possess an inlet, an apical trabecular component, and an outlet. This tripartite approach is also logical in the description of congenitally malformed ventricles. Rudimentary and incomplete ventricles lack one or more of its component parts, and are usually hypoplastic. The location and morphology of the rudimentary ventricles correlate with the disposition of the atrioventricular conduction system. CONCLUSIONS: Recognition of the ventricular components permits determination of ventricular morphology and guidelines for the location of the atrioventricular conduction axis. PMID- 9725441 TI - Which hearts are unsuitable for biventricular correction? AB - BACKGROUND: The surgical option of biventricular repair requires two ventricles, each fully capable of supporting the systemic or pulmonary circulation. The morphologic substrates that may preclude some hearts from biventricular repair need to be assessed. METHODS: Heart specimens were reviewed to assess the morphologic mechanisms that produce an unbalanced ventricular mass and to identify features that would, potentially, be a contraindication for biventricular repair. RESULTS: Hearts with solitary and indeterminate ventricles, and hearts with essentially solitary ventricles, often have associated abnormalities of venoatrial connections and arrangement of the atrioventricular valves. In the majority of hearts with univentricular atrioventricular connections, the rudimentary and incomplete ventricle of either right or left morphology may be too small to support either the systemic or the pulmonary circulation. Straddling with overriding of the atrioventricular valve, unbalanced atrioventricular septal defect, and gross hypoplasia of one of the ventricles in hearts with biventricular connections are other mechanisms producing ventricular imbalance, which could preclude biventricular repair. CONCLUSIONS: The morphologic mechanisms that result in ventricular imbalance are mainly related to the sizes and morphology of the ventricles, septal malalignment, valvar morphology, and component make-up of the ventricles. These features will influence decision-making in considering the option of biventricular repair. PMID- 9725442 TI - Echocardiography of hypoplastic ventricles. AB - BACKGROUND: Evaluation of hypoplastic ventricles with echocardiography requires an appreciation of the ultrasound methods used to assess ventricles of normal size. In this review, we present an overview of the most common techniques used to measure ventricular size, which may be analyzed as long- or short-axis dimension, area, volume, or mass. In addition to methods for evaluation, we review pertinent studies of sonographic evaluation of hypoplastic ventricles in consideration of their suitability for biventricular repair. METHODS AND RESULTS: Standard methods of volumetric and functional evaluation of the right and left ventricles are described, with a focus on their suitability for and applicability to the patient with a small ventricle. When applied to the patient with a hypoplastic ventricle, assessment may be more complicated in some respects, and requires consideration of functional characteristics of the ventricle itself, as well as the size and function of the corresponding atrioventricular valve. CONCLUSIONS: Echocardiography allows for excellent evaluation of ventricular size, morphology, and function. This holds true in patients with a hypoplastic ventricle as well, although the task is somewhat more complicated in such patients. PMID- 9725443 TI - Which two ventricles cannot be used for a biventricular repair? Echocardiographic assessment. AB - BACKGROUND: A variety of factors can influence the suitability of a congenitally malformed heart for biventricular repair, including size, morphology, function, and dimensions and function of the inflow and outflow, among others. Although certain features have been identified that may indicate a lower probability of successful biventricular repair, our ability to predict whether a particular patient will be able to tolerate completely separate in-series systemic and pulmonary circulations remains imperfect. METHODS AND RESULTS: In this review, we discuss the echocardiographic evaluation of various factors that can influence a patient's suitability for two ventricle repair. We call on our own experience, and illustrate our discussion with a number of echocardiographic images. CONCLUSIONS: In most cases, echocardiography allows for full assessment of the anatomic and functional features that influence whether a patient is a suitable candidate for biventricular repair. Although a number of indices have been developed for determining who can and cannot be expected to undergo successful two ventricle repair, there remains substantial room for progress in this area. PMID- 9725444 TI - Biventricular hearts not amenable to biventricular repair. AB - Many hearts, although considered morphologically biventricular, may not be candidates for a biventricular repair. Such patients are best placed on a Fontan algorithm. This article reviews in broad principles those hearts that, despite being biventricular, do not lend themselves to a two-ventricle repair. PMID- 9725445 TI - The pathology of subaortic obstruction. AB - BACKGROUND: In hearts having the atriums connected only to a dominant left ventricle, typified by double-inlet left ventricle but seen also in lesions such as tricuspid atresia, subaortic obstruction, when it exists, is usually found at the level of the ventricular septal defect when the aorta is supported by the rudimentary right ventricle. METHODS: Heart specimens were examined to determine the nature and position of the ventricular septal defect existing between dominant left and rudimentary right ventricles when the ventriculoarterial connections are discordant. RESULTS: Most commonly, the ventricular septal defect is positioned between the muscular apical trabecular septum and the muscular outlet septum. This type of defect is found not only in double-inlet left ventricle, but also in hearts with absence of either the right or left atrioventricular connection when the other atrium is connected to a dominant left ventricle, irrespective of the position of the rudimentary and incomplete right ventricle. Obstructive lesions within the aortic arch are commonly associated with restriction at the site of the ventricular septal defect. The atrioventricular conduction bundle takes a constant course relative to the margin of the septal defect. CONCLUSIONS: Because subaortic obstruction is almost always caused by a restrictive ventricular septal defect, relief of the obstruction can be achieved by surgical enlargement of the septal defect, bearing in mind the course of the atrioventricular conduction system. PMID- 9725446 TI - Subaortic obstruction and the Fontan operation. AB - Systemic outflow tract obstruction in the heart with a functional single ventricle promotes myocardial hypertrophy, and this has been shown to be an unequivocal risk factor for poor outcome at Fontan procedure. Such systemic outflow tract obstruction may be congenital or acquired. Those factors contributing to acquired systemic outflow tract obstruction in those patients with a double-inlet left ventricle, a rudimentary right ventricle, and a discordant ventriculoarterial connection include the size of the ventricular septal defect, previous pulmonary artery banding, and other volume unloading surgical procedures. Staging with a bidirectional cavopulmonary connection and construction of a proximal pulmonary artery-aortic connection or ventricular septal defect enlargement has neutralized this factor. PMID- 9725447 TI - Atrioventricular valve dysfunction: evaluation by Doppler and cross-sectional ultrasound. AB - BACKGROUND: An important factor in the management and outcome of patients with complex univentricular or partial biventricular repair is atrioventricular valve function. Cross-sectional and Doppler echocardiography are versatile tools for the evaluation of atrioventricular valve function. However, it is important to understand the physics and applications of this technology to appreciate the strengths and limitations of echocardiography in this application. METHODS AND RESULTS: In this review, we discuss the preoperative and intraoperative echocardiographic evaluation of atrioventricular valve function in congenital heart disease. The focus is on atrioventricular valve regurgitation, which is the most common type of dysfunction in patients with univentricular or partial biventricular heart disease. We emphasize an understanding of basic jet physics, as well as technical considerations in the evaluation of atrioventricular valve function, with illustrations from our own experience. CONCLUSIONS: Cross sectional and Doppler cardiac ultrasound is the optimal tool for evaluation of atrioventricular valve function in the current era. Although the issue of quantifying regurgitant jets is not yet fully resolved, echocardiography allows for complete qualitative assessment of the anatomic and functional features that influence the function of the atrioventricular valves. PMID- 9725449 TI - The physiology of the bidirectional cavopulmonary connection. AB - This article reviews the indications for the bidirectional cavopulmonary connection and demonstrates its efficacy in reducing mortality for the Fontan procedure. The indications for adding an additional source of pulmonary blood flow to the bidirectional cavopulmonary connection are discussed, but this issue remains controversial. Also unclear is whether the bidirectional cavopulmonary connection promotes symmetric growth of the pulmonary arteries, or whether growth of the left pulmonary artery is disadvantaged. Finally, systemic venous collateralization is a well-recognized sequel after cavopulmonary connection. The clinical implications of this collateralization are reviewed. PMID- 9725448 TI - Cavopulmonary anastomosis in staging toward Fontan operation: pathologic substrates. AB - BACKGROUND: A review of the history and practice of cavopulmonary connections in staging toward Fontan operation and the pathologic experience at Padua University is presented. METHODS: Gross and histologic assessment of the heart-lung blocks removed at autopsy in the cases in which the cause of death could be related to a dysfunction of the cavopulmonary anastomosis were performed. RESULTS: The main complications were distortion of the intraatrial tunnel and thrombosis of the pulmonary branches of the cavopulmonary anastomosis. CONCLUSIONS: The cavopulmonary anastomosis remains a widespread procedure, both as a preliminary step to a Fontan operation and as an integral part of a Fontan or modified Fontan procedure in all those cardiac malformations characterized by a hypoplastic right or left ventricle in which these ventricles are too small to support the whole circulation. PMID- 9725450 TI - Additional pulmonary blood flow with the bidirectional Glenn anastomosis: does it make a difference? AB - BACKGROUND: The bidirectional cavopulmonary shunt has become a mainstay in the palliation of patients with a functional single-ventricle heart. However, there remain a number of unresolved issues regarding this procedure, many of which concern the response of the pulmonary vasculature to this unique circulatory physiology. Among the issues of debate are the role and effects of an additional source of pulmonary blood flow. METHODS: Between January 1990 and April 1997, 160 patients underwent bidirectional cavopulmonary anastomosis. Median age at operation was 7.8 months, and age ranged from 24 days to 43 years. An additional source of pulmonary blood flow was included in 93 patients (58%). A retrospective review of our experience with this cohort was performed with a focus on the role of additional pulmonary blood flow. RESULTS: Eight patients (5%) died in the early postoperative period, and the overall early failure rate (death or take down) was 7.5% (n=12). Eleven other patients underwent early reoperation to decrease (n=8) or increase (n=3) the amount of pulmonary blood flow. Early survivors were followed up for a median of 23 months, during which time 5 patients died and 30 patients underwent Fontan completion. Including early and late mortality, actuarial survival rates at 1 and 2 years were 91% and 88%, respectively. CONCLUSIONS: The bidirectional cavopulmonary shunt is a useful procedure in the early or intermediate-term management of patients with a functional univentricular heart. However, there is much still to be learned about this unique physiologic system. The role of accessory pulmonary blood flow remains unclear, as does the use of the bidirectional cavopulmonary shunt as long term palliation. Pulmonary arteriovenous fistulas are a serious concern, especially in young patients with heterotaxy syndrome. PMID- 9725451 TI - Pathologic substrates for 1 1/2 ventricular repair. AB - BACKGROUND: The concept of "one and a half ventricular repair" relates to situations where one ventricle is capable of pumping one half of the circulation while the other ventricle is deemed inadequate and requires off-loading by means of a shunt. The inadequate ventricle is usually assigned the role of pumping the pulmonary circulation. The majority of hearts potentially amenable to this repair will have one large ventricle associated with a smaller and more-or-less rudimentary ventricle. METHODS: In this review, we focused on hearts in which the morphologically left ventricle will continue to support the systemic circulation. RESULTS: Among the hearts with univentricular atrioventricular connections, a few cases of classic tricuspid atresia and cases of double-inlet left ventricle coexisting with concordant ventriculoarterial connections would be suitable for incorporating the right ventricle into the pulmonary circulation. This procedure may be feasible in some cases of straddling and overriding tricuspid valve. Hearts with pulmonary atresia and intact ventricular septum display a wide range of sizes of the right ventricular cavity. Although biventricular repair is an option for those with good-sized cavities, patients with hypoplastic right ventricles may be candidates for one and a half ventricular repair. CONCLUSIONS: For the lesions reviewed, and many others, one and a half ventricular repair can be an option. PMID- 9725452 TI - A practical approach to 1 1/2 ventricle repairs. AB - BACKGROUND: Perioperative and long-term problems associated with the Fontan circulation are substantial. There has been an exploration of extending the limits of a biventricular ventricular repair by using a superior vena cava-to pulmonary artery anastomosis. This type of repair is known as a 1 1/2 ventricle repair. METHODS: Patients having defects of the pulmonary ventricle in size or function have undergone 1 1/2 ventricle repairs with or without creation of an atrial septal defect. Repairs with tricuspid z values as small as -10 and predicted pulmonary ventricular volumes as low as 30% have been reported. The 1 1/2 ventricle repair technique has also been used in special situations associated with an arterial switch or double switch procedure. RESULTS: Mortality has ranged from 0% to 12%. Complications have included persistent elevation of superior vena cava pressure, intermittent periorbital edema, and 1 superior vena caval aneurysm requiring takedown. There appears to be an increased risk of perioperative pleural effusions and chylothorax. Protein-losing enteropathy and chronic atrial arrhythmias have not been present. CONCLUSIONS: Successful 1 1/2 ventricle repairs have been reported for morphologically small or poorly functioning pulmonary ventricles and special situations. Intermediate-term follow up is favorable when compared with reported outcomes for the Fontan circulation. PMID- 9725453 TI - Total anomalous pulmonary venous connections and consideration of the Fontan or one-ventricle repair. AB - There is now a considerable literature that babies with right atrial isomerism have a poor outcome. The reasons for this are complex and multifactorial, but may be related at least in part to intrinsically small and abnormal pulmonary veins. We reviewed a series of consecutive patients seen at a single institution and found that babies with right atrial isomerism, severe pulmonary outflow tract obstruction or atresia, and total anomalous obstructed pulmonary veins had a grim outlook, especially those requiring operation in the neonatal period. Others have reported a similarly concerning outcome. PMID- 9725455 TI - Pulmonary arteriovenous malformations in and out of the setting of congenital heart disease. AB - BACKGROUND: Pulmonary arteriovenous malformations can occur in a variety of clinical situations, including liver disease, systemic disorders, or after palliation of congenital heart disease, with serious clinical consequences. METHODS: We reviewed the potential mechanisms of this condition, diagnostic tools, and clinical management. RESULTS: Contrast echocardiography is an important diagnostic modality, which has been shown to be more sensitive than pulmonary arteriography, especially when rapid contrast injection is used. The finding that pulmonary capillary vasodilation is observed in hepatopulmonary syndrome, in cirrhotic patients, and after congenital heart repair is strongly suggestive that an unidentified hepatic factor is involved in inhibiting the development of pulmonary arteriovenous malformations. CONCLUSIONS: Prompt detection and treatment of pulmonary arteriovenous malformations is of utmost importance, to prevent serious clinical consequences. It may very well be the case that the etiology of arteriovenous malformations is multifactorial. We are now investigating the role of alterations of gene expression in the vascular remodeling that results in formation of pulmonary arteriovenous malformations. PMID- 9725454 TI - Anomalous pulmonary venous return in the staged palliation of functional univentricular heart defects. AB - BACKGROUND: Bidirectional cavopulmonary shunt and Fontan repair are now commonly performed in patients with a variety of forms of complex single ventricle, including those with anomalies of systemic, pulmonary, or systemic and pulmonary venous return. These anomalies are ideally dealt with during bidirectional cavopulmonary shunt, thereby minimizing the complexity of the eventual Fontan procedure. METHODS AND RESULTS: Between March 1990 and March 1997, 32 patients with functional single ventricle and anomalous pulmonary venous return underwent operation at our institution. Five of 25 patients who underwent neonatal palliation died in the early postoperative period, all of whom had obstructed anomalous pulmonary venous return. Twenty-one patients have undergone bidirectional cavopulmonary shunt, including 7 in whom this was the primary palliative procedure. There was one early and two late deaths after the bidirectional Glenn procedure, two in patients with asplenia syndrome and none in patients with previously obstructed pulmonary venous return. Seven patients have undergone Fontan completion, 5 with an extracardiac conduit. There was one early death and one take-down to a classic Glenn shunt, both in patients who did not undergo the extracardiac conduit Fontan operation. CONCLUSIONS: Anomalous pulmonary venous return can significantly complicate the management of the single ventricle patient, with the major impact on survival coming in the neonatal period. Palliation with the aim of performing an extracardiac conduit Fontan procedure allows greater latitude and more streamlined management in this group of patients. PMID- 9725456 TI - Consensus statement on hypoplastic and rudimentary ventricles: morphology, angiography, and echocardiography. PMID- 9725457 TI - Application of fluorescence resonance energy transfer in the clinical laboratory: routine and research. AB - Fluorescence resonance energy transfer (FRET) phenomenon has been applied to a variety of scientific challenges in the past. The potential utility of this biophysical tool will be revisited in the 21st century. The rapid digital signal processing in conjunction with personal computers and the wide use of multicolor laser technology in clinical flow cytometry opened an opportunity for multiplexed assay systems. The concept is very simple. Color-coded microspheres are used as solid-phase matrix for the detection of fluorescent labeled molecules. It is the homogeneous assay methodology in which solid-phase particles behave similarly to the dynamics of a liquid environment. This approach offers a rapid cost-effective technology that harnesses a wide variety of fluorochromes and lasers. With this microsphere technology, the potential applications for clinical flow cytometry in the future are enormous. This new approach of well-established clinically proven methods sets the stage to briefly review the theoretical and practical aspects of FRET technology. The review shows various applications of FRET in research and clinical laboratories. Combination of FRET with monoclonal antibodies resulted in a boom of structural analysis of proteins in solutions and also in biological membranes. Cell surface mapping of cluster of differentiation molecules on immunocompetent cells has gained more and more interest in the last decade. Several examples for biological applications are discussed in detail. FRET can also be used to improve the spectral characteristics of fluorescent dyes and dye combinations, such as the tandem dyes in flow and image cytometry and the FRET primers in DNA sequencing and polymerase chain reactions. The advantages and disadvantages of donor-acceptor dye combinations are evaluated. In addition, the sensitivity of FRET provides the basis for establishing fast, robust, and accurate enzyme assays and immunoassays. Benefits and limitations of FRET-based assays are thoroughly scrutinized. At the end of the paper we review the future of FRET methodology. PMID- 9725458 TI - Morphological analysis of in situ hybridization signals in cervical intraepithelial neoplasia containing human papillomavirus type 16 or 18: relationship with histological grade and DNA content. AB - Among 345 lesions histologically defined as cervical intraepithelial neoplasia (CIN) examined by in situ hybridization (ISH) for the presence of DNA from human papillomavirus (HPV) types 6/11, 16, 18, 31, 33, and 51, a group of 69 lesions (41 low grade and 28 high grade) containing HPV 16 or 18 was further characterized with the following criteria: DNA ploidy and morphological patterns of ISH spots, i.e., punctate or diffuse throughout the nuclei corresponding to integrated or episomal state of HPV DNA, respectively. The highest percentage of aneuploid lesions, the highest diploid index values, and the highest proportion of CIN with punctate ISH signals were associated with high-grade lesions. In addition, punctate ISH signals were also most frequently found in aneuploid CIN. These results underline that punctate ISH signals considered as integrated HPV DNA were preferentially associated with aneuploid and high-grade lesions, and lead to suggest that this later criteria could be used to predict the evolution of a lesion towards malignancy. PMID- 9725460 TI - Detection of myeloperoxidase by flow cytometry in acute leukemia. AB - The value of flow cytometric detection of myeloperoxidase (MPO) in the differential diagnosis of acute leukemia was evaluated in 57 cases of acute leukemia and in 9 leukemia cell lines. Cells were fixed and permeabilized with Fix & Perm cell permeabilization kit at room temperature for 15 min each, and stained with anti-MPO monoclonal antibody (MPO-7) by direct immunofluorescence. One myeloid cell line, HL-60, was MPO-positive, while the other myeloid cell lines (KG-1, K-562, and MEG-01) as well as lymphoid cell lines (KM-3, NALM-6, Raji, REH, and T-ALL-1) were MPO-negative as previously described. Among acute leukemias, MPO was detected in 23 of 26 cases of acute myeloid leukemia (AML), 7 of 23 cases of B-lineage acute lymphoblastic leukemia (ALL), 1 of 6 cases of T lineage ALL (T-ALL), and 1 of 2 cases of acute unclassified leukemia (AUL). The intensity of MPO expression in 6 of 7 B-lineage ALL cases was weak compared with AML labeling. There was no detectable cytochemical MPO in the cells of ALL, AUL, or AML that stained negative for anti-MPO. No relationship between the expression of MPO and myeloid lineage surface antigens was observed in ALL. Three cases of MPO-positive ALL and AUL could be reclassified as biphenotypic leukemia according to the revised Catovsky scoring system. These results indicate that anti-MPO is an excellent marker for the diagnosis and classification of acute leukemia and can be reliably detected by flow cytometry. This rapid technique should be a valuable addition to routine immunophenotyping of acute leukemia. PMID- 9725459 TI - Comparison of routine flow cytometric DNA analysis of fresh tissues in two laboratories: effects of differences in preparation methods and background models of cell cycle calculation. AB - Routine flow cytometric DNA analysis was compared in two laboratories by using matched fresh-frozen breast cancer and soft tissue sarcoma biopsy specimens. Laboratory I applied the Vindelov preparation method and an exponential background subtraction algorithm in the cell cycle calculation. Laboratory II used the Formalin-protease preparation technique and the sliced-nuclei background model. The results of the ploidy analysis showed good agreement between the two laboratories; however, the results of the cell cycle analysis showed considerable systematic differences between labs. Laboratory I obtained significantly lower values of S-phase fraction and higher values of G2-phase fraction than laboratory II. To explain these discrepancies, the effects of differences in the preparation methods and background subtraction algorithms were studied. The Vindelov preparation method yielded higher debris and aggregation levels than the Formalin protease technique and tended to give higher %S and %G2 values. When the two background models were used in the same histograms, the exponential background model tended to give %S values distinctly lower than and %G2 values almost identical to those obtained with the sliced-nuclei algorithm. The sum of these effects accounts for the observed inter-laboratory discrepancies. Different from the sliced-nuclei fit, the exponential background fit often did not accommodate to the original data in the <2c histogram region and resulted in a considerable inter-operator variability of %S calculation in histograms with <5% S. When aggregate correction was added to the sliced-nuclei algorithm, the differences between %S values in histograms from the two laboratories almost disappeared. PMID- 9725461 TI - Quantitative fluorescence: to count or not to count. Is that the question? PMID- 9725462 TI - Development of midazolam sublingual tablets: in vitro study. AB - Midazolam is a benzodiazepine with short elimination half-life, used as induction or continuous agent for general anesthesia. At present, only injectable solution is available from French hospital pharmacies. The aim of the study is the development of 5 mg midazolam sublingual tablets to realize a short general anesthesia without intravenous or intramuscular injection. Incorporation of citric acid in the tablet formulation leads to an increase of dissolution rates of active drug, but a decrease of diffusion through lipid membranes is observed with 10 mg of citric acid when using the Dibbern's Resomat three phases apparatus. One explanation of this result is that midazolam (pKa = 6.1) in presence of 10 mg of citric acid is ionised. The ionised form, more hydrophilic, cannot cross the artificial lipid membrane and therefore the diffusion decreases. On the other hand, the decrease of diffusion's rate, when pH increases, is explained by the precipitation of midazolam at pH higher than 6. A compromise between dissolution and diffusion results leads us to choose the sublingual formulation containing 5 mg of citric acid per tablet. PMID- 9725464 TI - Gastric juice as a dissolution medium: surface tension and pH. AB - Aspirated gastric juice from eight patients was measured for surface tension, bile salt concentration and pH. Surface tension ranged between 35 and 45 mM/m, while pH was usually in the range 1-2 and bile salt concentrations were usually between 0 and 1 mM. No correlations were found between the three parameters. These findings suggest that the low surface tension of gastric juice cannot be attributed solely to refluxed bile salts. Once the source of the reduced surface tension is identified dissolution test media should be adjusted to represent these conditions. PMID- 9725463 TI - Influence of shaking and surfactants on the release of bsa from plga microspheres. AB - The aim of the present work was to study the release of a model protein, bovine serum albumin (BSA) encapsulated within biodegradable poly (D,L-lactide-co glycolide) (PLGA) microspheres prepared by a modified solvent evaporation method using a double emulsion. These microspheres were characterized for size, morphology, surface adsorbed protein, encapsulation efficiency and release kinetics. Two types of in vitro assays were developed to evaluate the influence of shaking and the addition of surfactants on the release profile of encapsulated protein. Scanning electron microscopy (SEM) observation showed spherical and smooth surface particles, with a mean particle size of 20 microm and an encapsulation efficiency of 81%. Surface associated protein was about 25%. The in vitro release profile showed a biphasic pattern described by means of a biexponential equation. There was an initial burst effect due to the release of the protein adsorbed on the microsphere surface and a sustained release phase due to protein diffusion through the channels or pores formed in the polymer coat. The release obtained profiles in static and dynamic assays showed statistically significant differences in the amount of the released protein, whereas the release rate was not affected. The burst effect was 28.30+/-1.63% and 35.20+/ 1.50% of the total encapsulated protein for the static and dynamic assays respectively. The addition of surfactants (SDS) to the release medium increased the rate and the amount of drug released. In both assays the value of the slow release rate constant, beta, was 0.029+/-0.002 days(-1) when the surfactant was added, and 0.017+/-0.0014 days(-1) in the samples without surfactant. It is believed that the surfactant leads to an increase in the microsphere surface polarity which allows channel and pore formation inside the polymer through which the protein diffuses easily. PMID- 9725465 TI - Effect of PEG 4000 on the dissolution rate of naproxen. AB - Naproxen is a nonsteroidal anti-inflammatory drug characterized by its low wettability and poor water solubility. Solid dispersions naproxen:PEG 4000 have been prepared in order to improve the solubility and dissolution rate of the drug, since these factors can be the limiting steps for absorption and bioavailability of poorly soluble drugs. X-ray diffraction analysis, infrared spectroscopy and differential scanning calorimetry detected no physico-chemical interaction between the drug and the inert carrier PEG 4000. The phase diagram of the naproxen-PEG 4000 system produced by DSC and hot stage microscopy is reported. The intrinsic dissolution rate of naproxen is calculated. The dissolution kinetics of solid dispersions prepared by the solvent and melt methods are compared with those of free drug and physical mixture. The studies were carried out at 37 degrees C and pH 1.2 according to the dispersed amount method. The dissolution profiles obtained indicate that a significant dissolution enhancement occurs with solid dispersions in comparison with the physical mixture. In addition, the physical mixture showed a dissolution rate higher than the free drug. Dissolution rate constants were determined by fitting the experimental data to the cube root function, to get straight line plots. PMID- 9725466 TI - Dissolution kinetics for coprecipitates of diflunisal with PVP K30. AB - Diflunisal is a nonsteroidal anti-inflammatory drug that is poorly soluble in water. The present study describes the formulation of solid dispersions of the drug designed to increase its solubility. X-ray diffraction and DSC were used to examine the physico-chemical characteristics of solid dispersions of diflunisal and polyvinylpyrrolidone (PVP) prepared by the solvent method, using percentage proportional compositions ranging from 20:80 to 50:50. X-ray diffraction analysis detected that diflunisal is present in solid dispersions in crystalline or amorphous state depending on the PVP content. The thermal behavior of diflunisal observed in the DSC curves of solid dispersion systems, was attributed to a solid state interaction. The increased release of the PVP-drug dispersion as compared to the PVP-drug physical mixture was attributed to the formation of a complex resulting from the interaction of the drug and the polymer. PMID- 9725467 TI - Preparation and dissolution rate of gliquidone-PVP K30 solid dispersions. AB - Solid dispersions of gliquidone in PVP K30 were prepared by the solvent method. These dispersions were characterized using X-ray diffraction. In comparison with the drug alone, the physical mixtures and even more the solid dispersions showed an increase in the dissolution rate. Moreover these solid dispersions were stable during storage. PMID- 9725468 TI - Pharmacokinetic examination of antipyrine passage through the placenta and the small intestine in rats. AB - The placental and small intestinal barriers, though obviously different, show many functional as well as morphological similarities. When the surface area of both barriers in man was recalculated to a unit of body weight, nearly identical values (2.71 and 2.86 m2/kg of body mass, respectively) were obtained. The aims of the present study were (1) to compare mutual permeability of these two barriers to antipyrine (AP), and (2) to describe pharmacokinetics of AP in pregnant and non-pregnant rats. In placental studies AP showed that its rapid transfer through the placenta (k(tr) = 0.046 min(-1)) was governed by the mechanism of passive diffusion. In the closed circuit, FMCR(eq) was 1.085, t(eq) was 112.10 min and k(eq) was 0.020 min(-1). Absorptive studies performed on the rat small intestine indicated an identical mechanism of drug transport. The apparent first-order absorption rate constant of AP was 0.479 min(-1), and Tmax was 8.95 minutes. Differences in AP pharmacokinetics between pregnant and non pregnant rats were significant during the distribution phase (t(1/2) = 3.78 and 5.87 min, respectively), whereas the elimination phase was unaffected. AP has been demonstrated, as expected, to be an excellent marker for drug transport studies through different body barriers. PMID- 9725469 TI - Distribution of deramciclane (EGIS-3886) in rat brain regions. AB - The time related distribution and pharmacokinetics of double-labelled EGIS-3886 (EGIS-3886-phenyl-14C and -ethyl-3H) were studied in the plasma, hypophysis and 14 cerebral regions, including the spinal cord of the rat after a single oral treatment (acute experiments) and after repeated administration of one dose daily for six days (subacute experiments). The tissue levels of EGIS-3886 (deramciclane) were calculated from the simultaneously determined dpm values and the specific activities of the two radioisomers present in the dose administered. EGIS-3886 was rapidly absorbed from the gastrointestinal tract (t(max)=1.0 h). The concentration-time curves in the tissues can be described by a two compartment open model. The 3H-activity could be measured during the whole period of the acute experiment (96 h), whereas 14C-radioactivity fell below the detection limit within 24 h. The AUC(0-96) values for 3H were 10 to 15 times higher than that for 14C. In all samples examined, on the concentration time curves a peak characteristic of enterohepatic cycle can be seen at 12 h. The studies indicated that intact molecules entered brain tissues from the circulation. The results of the subacute experiments indicate that the 14C labelled EGIS-3886, or its metabolite(s) carrying the tracer, reach an equilibrium as early as on the second to third day, whilst the level of 3H radioactivity continually increases during the six days of repeated administration. In the subacute experiments the peak concentrations were reached at 0.5 h after the final treatment. However, their values for 3H were higher than in acute experiments. The last tendency was not observed in the case of 14C tracer. The AUC values of 3H-labelled EGIS-3886 determined in subacute experiments predominated over 14C; the ratios were 50 to 60 in all brain regions. The enterohepatic cycle, seen after a single dose, also operated after repeated dosage. The time related concentrations of EGIS-3886 in the hypophysis were at least two times higher than that in the plasma and the brain tissues. No significant difference was seen in the concentrations of EGIS-3886 in the symmetrical (left and right) regions of the brain. PMID- 9725471 TI - Catechol is the major product of salicylate hydroxylation in 1-methyl-4 phenylpyridinium ion treated rats. AB - Salicylate hydroxylation using hydroxyl free radicals results into formation of 2,3-dihydroxybenzoic acid, 2,5-dihydroxybenzoic acid and catechol. Inspite of the fact that in vitro experiments have shown that catechol is a minor product, we have shown by these in vivo studies that it is a substantial product. Since catechol as well as 2,3-dihydroxybenzoic acid have not been found to be produced enzymatically from salicylates, they appear useful as in vivo indicators for monitoring hydroxyl free radicals. Administration of 1-methyl-4-phenylpyridinium ion (MPP+) to rat striatum using microdialysis results into the formation of hydroxyl radicals. Salicylate perfusion enables the estimation of the three derivatives cited above. They increased significantly after MPP+ administration in comparison to the baseline values, with catechol being the most significant. The maximum amounts were achieved 60 min after MPP+ administration, and the mean percentage increase at this time point were 83.1% for 2,3-DBA (n = 6, P = 0.005), 81.25% for 2,5-DBA (n = 6, P = 0.011) and 1228.8% for catechol (n = 4, p = 0.00008). MPP+ caused substantial decrease of dopamine metabolites. Dihydroxyphenylacetic acid decreased to 13% and homovanillic acid to 11.4%. We conclude that catechol is an important indicator of hydroxyl free radical formation in this animal model which is well suited to study the role of free radicals in Parkinsonism. PMID- 9725470 TI - In vitro study of the binding of doxorubicin to heart. PMID- 9725472 TI - Pharmacokinetic parameters of netilmicin and protective effect of piperacillin regarding nephrotoxicity caused by netilmicin. AB - The pharmacokinetic interaction of Netilmicin and Piperacillin has been studied as well as the potential protective effect that Piperacillin exert on nephrotoxicity caused by Netilmicin, when both antibiotics are administered to rabbits by single and multiple dosage regimens. Netilmicin was administered at a dose of 7 mg/kg and 12 h interval, which allometrically correspond to 5 mg/kg at 24 h interval for men. Piperacillin was administered at a dose of 280 mg/kg at 12 h interval (the total number of doses of both antibiotics was 20). After single and multiple dose regimens plasma level curves of Netilmicin and renal concentration were determined using an HPLC technique. Besides that, an histologic study was carried out by electronic microscopy to determine the renal damage. A significant variation of some pharmacokinetic parameters of Netilmicin such as Vc and t(1/2) was observed when Netilmicin is administered together with Piperacillin; a similar modification in the renal accumulation and renal damage caused by Netilmicin was shown. PMID- 9725473 TI - Robust methods in bioequivalence assay; preliminary results. AB - The aim of this study is to compare four statistical methods for outlier identification in Bioequivalence tests. The methods are based in four confidence intervals, 'parametric','non- parametric','robust with the asymptotic distribution of the M-estimator' and 'robust with the bootstrap distribution'. The drug we used was Diltiazen, in a two sequence randomized crossover study design. The pharmacokinetic parameters measured were the area under the plasma concentration curve (AUC), and the peak drug concentration (CMAX). Time to peak drug concentration (TMAX), was not used here in order to separate the efficiency of the methods from the efficiency of the measurements. The methods were applied to simulated and experimental data. We made two simulations, one with normal data and another one with outliers data. When simulating normal data all methods showed similar profiles and high power. On the contrary, when simulating experiments with outliers data, the parametric method showed low power, whereas robust methods showed just a slight decrement in power. When we analyzed experimental data of AUC, if we used the parametric method (recommended by U.S.P), we were not able to conclude Bioequivalence, but with the other methods, this was possible. This disagreement between parametric and robust procedures was a sign of outliers data. We conclude that the robust methods in bioequivalence assays help us in the identification of outliers as observations with weight equal zero. PMID- 9725474 TI - Average parameters as a trend to reduce the residual variability in bioequivalence trials. AB - Bioavailability and bioequivalence evaluations of drug products carried out using the experimental maximum concentration (Cmax) and the experimental time to reach Cmax (Tmax), are not advisable for slow-release formulations and for trials performed with saliva as biologic fluid. When slow-release curves are considered the drug concentration profiles usually show multiple peaks, making it difficult to compute a Cmax,Tmax value. The saliva profile throughout time shows a high variability observed as more than one peak in the saliva concentration versus time curves. In both cases, even if there is a major peak, when the statistical analysis of the data is performed, an important variability in Cmax results in high values in the residual variance of the ANOVA test. Consequently, the power of the bioequivalence test decreases and sometimes it is not possible to conclude on bioequivalence. The average concentration (Cav), the average maximum concentration (Cmax,av) and Cmax,av/Cav x 100 (%Cmax,av) are proposed in this paper as possible parameters in order to evaluate the profile of the concentration-time curves, as they reduce the residual variability in bioequivalence studies. PMID- 9725475 TI - Average parameters in bioavailability studies: an application to slow-release amitriptyline formulation. AB - In order to assess the extent and the rate of absorption in bioavailability studies, area under the curve (AUC), experimental maximum concentration (Cmax) and experimental time to reach Cmax (Tmax), are used. But when slow-release formulations are considered, the drug concentration-time curves usually show multiple peaks, and it is difficult to compute a Cmax and Tmax value. In case a Cmax value is computed, important variability in this parameter results in high values in the residual variance of the ANOVA test. So in order to decrease the high variability, average parameters: average concentration (Cav), average maximum concentration (Cmax,av) and Cmax,av x 100/Cav (%Cmax,av), are proposed. These new parameters were applied in a bioavailability study of slow-release amitriptyline formulation. PMID- 9725476 TI - Which bioequivalence study for a racemic drug? Application to milnacipran. AB - Milnacipran, a new non tricyclic antidepressant drug, is a racemic mixture (F2207) composed of two enantiomers: F2695 and F2696, both demonstrated to be active. A randomized open label, single-dose latin square study was undertaken in 24 healthy volunteers to compare, based on racemate data, the relative bioavailability of two new formulations to that of a reference formulation. Later on, as suggested by actual regulatory trend, analysis was carried out on enantiomer data, although in a supportive way. Bioequivalence was assessed on calculation of 90% confidence intervals for log-transformed Cmax and AUC(0 infinity) and on Wilcoxon test for Tmax with a 5% level of significance. Based on racemate data, both test formulations were demonstrated to be equivalent to the reference capsule in terms of Cmax and AUC-(0-infinity). Differences in Tmax reached statistical significance, although their mean magnitude was small, and probably not relevant when related to antidepressant long-term therapy. When considering the test capsule - reference capsule comparison, the equivalence demonstrated for the racemate reflect that of each enantiomer. On the contrary, equivalence between the test tablet and the reference capsule demonstrated for the racemate, is not supported by both enantiomers as Cmax of F2696 fails to reach bioequivalence criteria, making more uncertain the conclusion of bioequivalence. From this experience, it seems than when equivalence is demonstrated close to the limits for the racemate, it is difficult, especially for a low variability drug such as milnacipran, to comply with equivalence criteria for both enantiomers. PMID- 9725478 TI - Relative bioavailability of different oral sustained release oxprenolol tablets. AB - The bioequivalence of oral dosage forms of oxprenolol was assessed in a triple crossover study on two groups of 12 volunteers each. Single 160 mg doses of oxprenolol hydrochloride were given after an overnight fast of either oxprenolol sustained-release tablets in a megaloporous system, a hydrophil matrix and Slow Trasicor (Ciba-Geigy) in the first group, or floating slow-release tablets administered with food or in absence of food, and rapid release Oxprenolol (Terapia, Cluj-Napoca) tablets, in the second group. Serum oxprenolol concentrations were measured by a gas chromatographic method. Pharmacokinetic parameters which describe bioavailability and general kinetic behavior of the drug were calculated from individual serum profiles. They were subjected to statistical analysis (paired Student's t test, p < 0.05). The customary bioequivalence criterion was used: 0.8 < parameter ratio(tested/standard) < 1.2. Megaloporous tablets showed bioequivalence with the reference sustained release product Slow-Trasicor. Hydrophil tablets showed moderate sustained-release characteristics. Floating tablets showed significantly greater oxprenolol absorption when taken with food and were non-bioequivalent with floating tablets without food, as well as with the reference rapid release tablets, of oxprenolol. However, fasting tablets were bioequivalent to the Slow-Trasicor product, when taken with food. PMID- 9725477 TI - Comparative bioavailability of two oral formulations of ipriflavone in healthy volunteers at steady-state. Evaluation of two different dosage schemes. AB - Ipriflavone (IP) is an isoflavone derivative with antiosteoporotic activity. This drug is extensively metabolized in humans and only negligible concentrations of unchanged IP can be detected in plasma. Metabolites M1 and M5 are predominant, while met abolites M2 and M3 are detected in minor amounts. The aim of this study was to compare the bioavailability of IP and its metabolites M1, M2, M3, and M5 at steady-state after administration of 200 mg tablets three times daily and 300 mg Scherer capsules twice daily during meals. IP plasma levels were below the limit of quantitation in 6 subjects out of 12 after administration of IP 200 mg tablets. On the other hand, after administration of the Scherer capsules IP plasma levels were quantifiable in all the volunteers. As regards IP metabolites, a mean increase in bioavailability, equal to 35%, was observed after administration of the Scherer capsules. Plasma level fluctuations, reflecting changes in absorption rate at steady-state, remained unvaried. The good bioavailability and fluctuation indexes of the Scherer capsules permit a simplification of the dosage scheme, reducing the daily administrations from three times to twice daily, thus improving the patients' compliance. In clinical practice this characteristic is not negligible, con sidering the mean age of the patients and the long-term treatment. Due to the high therapeutic index of IP, the increase in bioavailability does not cause any risk of accumulation or overdosage. PMID- 9725479 TI - Comparative bioavailability study of codeine and ibuprofen after administration of the two products alone or in association to 24 healthy volunteers. AB - The study objective was to compare the bioavailability of codeine and ibuprofen after oral administration of the two drugs alone or in association. The study was performed in three different periods, each separated by a wash-out of 6 days. Plasma concentrations were measured in 24 healthy volunteers after administration of a single oral dose of codeine phosphate (25 mg) and/or ibuprofen (200 mg). Codeine and ibuprofen assays were performed using two different HPLC methods. The relative bioavailabilities of codeine and ibuprofen (alone or in association) were 106 +/- 24% (mean +/- sd) and 101 +/- 19%, respectively. The results obtained demonstrated that bioavailabilities of codeine and ibuprofen were not modified when the two drugs were administrated alone or in association. PMID- 9725480 TI - Biopharmaceutical evaluation of time-controlled press-coated tablets containing polymers to adjust drug release. AB - This paper deals with press-coated modified release tablets in which the drug dose is situated in the core or is divided between the core and the coat. The coat contains polymer (sodium alginate or hydroxypropylmethyl cellulose, HPMC) to control drug release. The main objective was to investigate how the pharmacokinetic profile of the model drug could be modified by altering the proportion of the drug between the core and the coat. The effect of the amount of the polymer in the coat was also studied. Bioavailability tests were carried out on healthy volunteers. In the absorption curves of the tablets containing 50%, 67% and 80% of the drug in the core and 180 mg HPMC in the coat a bimodal profile was observed. No bimodal release pattern in the in vitro dissolution studies was found. If the whole dose was incorporated in the core the absorption curve has only one clear t(max) value at about 10 h. Doubling the amount of HPMC in the coat dramatically decreased drug absorption. It was concluded that, if a slightly reduced t(max)-value was required, the viscosity grade of HPMC used should be lowered. PMID- 9725481 TI - Food effect on the oral bioavailability of Manidipine: single dose, randomized, crossover study in healthy male subjects. AB - The effect of food on the oral bioavailability of a manidipine 20 mg tablet was studied after a single administration in 12 male healthy subjects. The clinical trial was conducted as an open, randomised, crossover study. In two different administration sessions, the subjects received a 20 mg manidipine tablet either in the fasting state or after a standardized breakfast. Plasma samples were collected before and at different times after each administration for up to 24 h. The concentrations of manidipine and its pyridine metabolite (M-XIII metabolite) were determined by HPLC with coulometric detection. The tolerability of manidipine was good. Only two cases of mild headache, one with each treatment, were reported. Food significantly improved the absorption, with an increase in AUC from 19.1 to 27.2 ng.h/ml (geometric mean, p<0.01) but did not modify the rate of absorption (tmax unchanged, median = 1.5 h). Peak plasma concentration was also increased (from 6.2 to 7.8 ng/ml), but the difference was not statistically significant (p=0.18). Other pharmacokinetic parameters (apparent elimination half-life and mean residence time) remained unchanged. The increase in bioavailability of manidipine administered with food is related to its high lipophilicity and may be explained through a solubilization effect produced by food and bile secretions. PMID- 9725482 TI - Is one paracetamol suppository of 1000 mg bioequivalent with two suppositories of 500 mg. AB - A common belief is that one tablet or suppository containing, e.g. 100 mg of a drug can be substituted, without any changes in the therapeutic effect, with two units of the same brand containing 50 mg of the drug. In the present study a single dose of paracetamol was administered to healthy volunteers as (a) two tablets of 500 mg, (b) two suppositories of 500 mg, and (c) one suppository of 1000 mg. There were statistically significant differences in all bioavailability parameters (t(max), C(max) and AUC) between the three treatments. The relative bioavailability of the 500 mg suppositories was 77% and that of the 1000 mg suppositories 66%. The absorption rate from suppositories was markedly lower than from the tablets. Especially low absorption rate was obtained with the suppository of 1000 mg. The two strengths, although having the same trade name, were not therefore bioequivalent. PMID- 9725483 TI - LIMS: from theory to practice. AB - This paper gives a definition and some basic knowledge about Laboratory Information Management Systems (LIMS) as well as their impact on the organisation, the laboratory and the co-workers. The major advantages and disadvantages of LIMS are pointed out. Two practical experiences are described. The first is related to an in house development of a PC based system which has to integrate a Vax VMS system (Multichrom) and PC based analytical and analysis softwares. The second experience is dealing with the selection and implementation of a commercial package in a pharmacokinetic laboratory. In both cases the human and time aspects were important. PMID- 9725484 TI - Bioavailability of the iron formulated as natural ferric protein (TM/FMOA) and natural ferric protein + folic acid (TM/FMOA+FOL). AB - Numerous oral iron preparations are available for the treatment of iron deficiency anemia but only very few studies have been designed to measure the bioavailability of iron preparations. The aim of this assay is to determine the bioavailability of iron formulated as natural ferric protein (TM/FMOA)(FMOA=Ferrimannitol ovalbumin) and natural ferric protein with folic acid (TM/FMOA+FOL) in healthy and anemic rats. Determination of the bioavailability of iron has been carried out by studying the serum level-time curves after i.v. administration of a single dose of 1 mg iron/kg in the form of ferrous sulfate, and after oral administration of 2 mg of iron/kg in the form of TM/FMOA or TM/FMOA+FOL. The Cmax values obtained after oral administration of TM/FMOA and TM/FMOA+FOL are greater in the case of anemic rats, while the tmax values are similar in both types of animals. The bioavailability (F) is greater for the anemic rats (80% approx), and the presence of folic acid does not appear to influence the bioavailability of the iron. This bioavailability data with TM/FMOA can predict good results in further experiments in human beings with iron deficiencies. PMID- 9725485 TI - Amiloride pharmacokinetics in rat. AB - The kinetics of amiloride was investigated in plasma, urine, faeces and tissues of rats after oral (10 mg/kg) and i.v. (10 mg/kg bolus and 35 microg/h for 4-days infusion) administration. Initially the experimental data were analyzed by a multiexponential model, then a compartmental model was developed to describe the drug kinetics in plasma, urine, faeces and tissues after the i.v. bolus and the oral administration simultaneously. Aim of the model was also to predict the drug kinetics in plasma and tissues of rats after continuous i.v. infusion. The results of the prediction and the discrepancies between prediction and observed data allowed a deeper insight into the pharmacokinetics of amiloride. PMID- 9725486 TI - 14C-NaVP and 14C-PEV repeated dose study in rat. Pharmacokinetic study in rats after repeated oral administrations of 14C-valproic acid sodium salt and 14C valproic acid pivaloyl oxymethyl ester. AB - The absorption, excretion and tissue distribution of radioactivity after repeated oral equimolar doses of 14C-valproic acid sodium salt (NaVP) or 14C-valproic acid pivaloyl oxymethyl ester (PEV) was investigated in male rats treated once a day for 14 consecutive days. The 14th day plasma time-course of radioactivity after PEV administrations was characterised by a slow absorption rate with a delayed peak (tmax 2 h, Cmax 7.52 +/- 1.35 microg eq./ml), followed by a plateau lasting up to 8 h. After NaVP treatment, the main peak of radioactivity was observed 0.5 h after administration (Cmax 8.30 +/- 1.26 microg eq./ml) followed by a secondary peak due to biliary enterohepatic recycling. Starting from 4 h onwards, radioactivity levels after PEV treatment were higher than those after NaVP (AUCtau = 113.3 h.microg eq./ml after PEV vs 71.9 h.microg eq./ml after NaVP), but concentrations declined with similar terminal half-lives (52.8 h for PEV and 49.7 h for NaVP). Radioactivity recovered (0-432 h interval) in urine accounted for 79.3% (PEV) and 56.1% (NaVP) while, in faeces accounted for 9.1% (PEV) and 26.1% (NaVP) of total administered dose (14 days). The difference is attributable to a higher excretion of radioactivity in the bile for NaVP. The missing fraction in the total radioactivity balance is probably excreted in expired air, as observed in single dose studies. Radioactivity excreted in bile (0-8 h interval of the last 14th day) accounted for 5.1% (NaVP) and 0.23% (PEV) of the total administered dose (14 days). A possible explanation of this difference may be a different metabolism pattern for the two compounds. The negligible biliary excretion observed after PEV administration is probably due to an inhibition of the glucuronation of valproic acid (or other metabolites) caused by the pivalic acid. Due to the presence of the enterohepatic recycle, the radioactivity levels in intestine, 0.5 and 2 h after administration, were higher after NaVP administration. According to higher plasma levels, the radioactivity concentrations in liver, kidneys and some fat tissues were found to be slightly higher after PEV administration. At 120 h after the last treatment of both compounds, relevant tissue concentrations were observed in mesenteric lymphnodes, perirenal and brown fat. The tissue-plasma radio activity ratio appeared quite similar for the two compounds. PMID- 9725487 TI - Pharmacokinetic study in rats after single intravenous and oral administrations of [14C]-ITF-296. AB - Pharmacokinetics of [14C]-ITF-296 and its metabolites, ITF-1124 and ITF-1577, were studied in rats after a single intravenous (2.5 mg/kg) and oral (10 mg/kg) administration. Radioactivity was measured by LSC while unchanged drug and its metabolites in plasma were assayed by an HPLC-UV method. The absorption of [14C] ITF-296 after oral administration is practically complete. Elimination of radioactivity occurs mainly in urine (higher than 80%) and the recovery of the dose (higher than 95%) takes place up to 96 h after both treatments. Both by i.v. and p.o. route the results show that the radioactivity is largely excreted in the bile and reabsorbed in the intestine. The tissue distribution study indicates that there is no accumulation or localization of radioactivity in the major organs or blood and no radioactivity levels are found 96 h after either treatment. In addition, whole body autoradiography confirms the tissue distribution pattern, showing no differences between albino and pigmented rats. PMID- 9725489 TI - Pharmacokinetics of Camonagrel in experimental animal: rat, rabbit and dog. AB - Camonagrel is a novel selective thromboxane synthetase inhibitor. The aim of this study was to determine its main pharmacokinetic parameters in rats, rabbits and dogs after intravenous and oral administration at doses of 10 mg kg(-1). Plasma and urine concentrations of camonagrel were analyzed by HPLC with UV detection. Pharmacokinetics of camonagrel was generally fitted to a two-compartmental model and the values which defined the absorption process were: Cmax = 15.96 microg.ml( 1), Tmax approximately 0.33 h, AUC(0-infinity) (oral) approximately 12.45 microg x h x ml(-1) (rat, n=3 per pont); Cmax approximately 2.04 mg x ml(-1), Tmax approximately 1.50 h, AUC(0-infinity) (oral) approximately 4.85 microg x h x ml( 1) (rabbit, n=3); Cmax approximately 18.60 microg x ml(-1), Tmax approximately 0.44 h, AUC(0-infinity) (oral) approximately 13.40 microg x h x ml(-1) (dog, n=4). The more representative values in the distribution and elimination phase were: protein binding rate approximately 80% in the three species ("in vitro" experiment); t(1/2beta) approximately 0.22 h (rat, i.v.), = 0.28 h (rabbit i.v.) and approximately 0.45 h (dog i.v.); CI approximately 635.73 ml x h(-1) (rat i.v.), approximately 448.26 ml x h(-1) (rabbit i.v.) and approximately 463.8 ml x h(-1) (dog i.v.). The absolute bioavailability of camonagrel was approximately 79.1% in rat, approximately 21.7% in rabbit and approximately 59.3% in dog. Available elimination data in rat indicated that Camonagrel was mainly excreted in urine (approximately 80%) as unchanged drug. An unknown minor metabolite (approximately 10%) was observed only after oral dosing. Finally, the main pharmacokinetic parameters of camonagrel in rats, rabbits and dogs are presented, which allow to define its absorption, distribution and elimination processes in these species. PMID- 9725488 TI - Pharmacokinetic study in dogs and monkeys after single intravenous and oral administrations of [14C]-ITF-296. AB - Pharmacokinetics of [14C]-ITF-296 and its metabolites, ITF-1124 and ITF-1577, were studied in dogs and monkeys after a single intravenous (2.5 mg/kg) and oral (10 mg/kg) administration. Radioactivity was measured by LSC while unchanged drug and its metabolites in plasma were assayed by an HPLC-UV method. The absorption of [14C]-ITF-296 after oral administration is practically complete both in dogs and in monkeys. The determination of unchanged drug and its metabolites shows quite a similar profile in dogs and monkeys for ITF-296 and ITF-1124 and a different time-course for ITF-1577. Elimination of radioactivity occurs mainly in urine (namely 70-80%) for both species and the recovery of the dose (higher than 90%) takes place up to 96 h after both treatments. PMID- 9725490 TI - Pharmacokinetics of midazolam and its main metabolite 1-hydroxymidazolam in intensive care patients. AB - The pharmacokinetics of midazolam and of its main metabolite, 1-hydroxymidazolam, were investigated in intensive care patients after intravenous bolus of 0.2 mg/kg followed by a 0.1 mg/kg/h intravenous infusion of midazolam over 2 hours. A wide interpatient variability of the main pharmacokinetic parameters of midazolam was found. The mean values of elimination half life and volume of distribution, 4.5 +/- 5.4 h and 1.7 +/- 0.7 l/kg respectively, were higher than those reported in healthy subjects. Total plasma clearance was significantly increased in patients taking drugs that induce hepatic metabolism. Significant concentrations of the unconjugated form of 1-hydroxymidazolam were recovered in plasma. The volume of distribution and the elimination half life of the metabolite were higher than those of the parent drug. These results show that 1-hydroxymidazolam might contribute to the pharmacodynamic effect of midazolam and consequently must be taken into account during pharmacokinetic and pharmacodynamic studies. PMID- 9725491 TI - Stereoselective metabolism of mexiletine in Chagasic women with ventricular arrhythmias. AB - Following a week of racemic mexiletine HCl at 200 mg tid (2x100 mg capsules), stereoselective aliphatic hydroxylation was studied in eight Chagasic women with chronic ventricular arrhythmias (52-67 yrs) with no history of renal or hepatic diseases. Blood samples were collected at dose interval up to 24 h of drug administration. Plasma concentrations of R(-) and S(+) mexiletine (MEX) and its metabolite hydroxymethylmexiletine (HMM) were determined by HPLC after derivatization with chiral reagent. The differences between R(-) and S(+) enantiomers were compared by paired t-test. Results are mean (95% CI). The following differences (p < 0.05) between R(-) and S(+) enantiomers, respectively, were found: MEX AUCss(0-8) 2.34 (1.84-2.85) vs 2.55 (1.97-3.13) microg.ml(-1) x h(-1); MEX CL/f 11.27 (7.77-14.77) vs 10.46 (7.18-13.74)ml.min(-1).Kg(-1); HMM Cmax 38.26 (24.3-52.22) vs 16.73 (10.1-23.29)ng.ml(-1); HMM Tmax 4.71 (2.67-6.76) vs 3.29 (1.24-5.33) h and HMM AUCss(0-8) 253.50 (165.39-341.61) vs 103.70 (69.51 137.90)ng.ml(-1).h(-1). The AUCss(0-8) ratio R(-)/S(+) for MEX was 0.93 (0.87 0.98) while for HMM was 2.50 (2.16-2.85). Distribution of MEX and HMM enantiomers were not significantly different. In this study we demonstrate that kinetic disposition of mexiletine exhibits stereoselectivity in vivo and that aliphatic hydroxylation is favored for R(-) mexiletine in Chagasic women with ventricular arrhythmias. PMID- 9725492 TI - Technetium-99m labelling of human immunoglobulin and preliminary clinical evaluation in tumour patients. AB - In the present investigation we have human immunoglobulin labelled with 99Tc(m), applying two different systems. The radiochemical characteristics of the labelled antibody were studied by conventional, radioanalytical methods. Further on, pharmacokinetics of this 99Tc(m)-labelled biomolecule were investigated, by i.v. administration in women, checked for tumours of the ovaries, uterus and cervix. Scintigraphic findings were compared to the results of other imaging techniques (CT, US, X rays), as well as to surgical findings. Our studies indicated that 99Tc(m)-human immunoglobuline can be applied successfully for the scintigraphic detection of several malignant and benign tumours of the female genital system. Tumour accumulation is probably due to the activation of particular cells, as macrophages and lymphocytes, responsible for inflammatory and immunological responses. PMID- 9725493 TI - Pharmacokinetics of milnacipran in liver impairment. AB - The pharmacokinetics of single 50 mg oral and intravenous doses of milnacipran, a new non tricyclic antidepressant drug, were compared in 11 chronic liver impaired (LI) subjects and in 6 control subjects. Hepatic impairments, classified according to the PUGH scale were moderate (1 grade A), intermediate (6 grade B) and severe (4 grade C). Concentrations of unchanged drug and its conjugated form (its main metabolite) were measured in plasma and urines. In control subjects, milnacipran present high absolute bioavailability (mean value of 90%). Around 50% of the dose are excreted in urines as unchanged, while around 14% are excreted as glucuroconjugate. The remaining is composed of free and conjugated phase I inactive metabolites. Administration of milnacipran in LI subjects results in non significant changes in its pharmacokinetics, although its variability is increased. Unchanged drug exposure is not modified in LI subjects, while plasma levels of the conjugate are slightly decreased compared to the control group. This could either be due to a slight reduction in the conjugation process, or to a change in the distribution of the drug as urine excretion of both unchanged and conjugated forms are not modified compared to the control group. A few LI subjects present supra-bioavailability resulting in higher drug exposure after oral administration than after intravenous infusion. These modifications are not clinically relevant as drug exposure of the parent drug is not modified. As the unchanged drug is the only compound responsible for the activity of milnacipran, no dosage adjustment is needed in patients presenting liver impairment. PMID- 9725494 TI - Pharmacokinetics of milnacipran in renal impairment. AB - The pharmacokinetics of a single 50 mg dose of milnacipran, a new non tricyclic antidepressant drug, were compared in 8 chronic renal failure subjects (Clc(reat) between 9 to 84.5 ml.min(-1)) and in 6 healthy volunteers. Concentrations of unchanged (F2207 racemate and F2695 and F2696, enantiomers) and glucuroconjugated drug (main metabolite) were measured using HPLC and GC-MS. As for drugs mainly eliminated via renal route, the pharmacokinetics of milnacipran were markedly affected by impaired renal function with the elimination half-life of severely impaired subject being approximately three times that of the control group. Milnacipran apparent total clearance and renal clearance were positively correlated with glomerular filtration rate, while non-renal clearance and apparent volume of distribution were unaffected by renal impairment. Plasma concentrations of the glucuroconjugate were gradually increased in plasma, while its total urine excretion remained unchanged. As for the racemate, pharmacokinetics of each enantiomer were modified by renal failure, although, as predictable from its higher renal clearance value, it was more marked for F2696 than for F2695. Considering that modifications were shown to be proportional to the degree of renal impairment and that milnacipran presents low variability, the necessary dose adjustment is therefore easy to predict. PMID- 9725495 TI - Distribution of fenofibric acid in lipoprotein fractions of patients. AB - The antidyslipidemic agent fenofibrate (procetofen) is hydrolysed in vivo to its main active metabolite--fenofibric (procetofenic) acid. This metabolite is usually determined in pharmacokinetic studies, because plasma concentrations of fenofibrate are practically undetectable. Presented study is focussed on the distribution of fenofibric acid into lipoprotein (VLDL, LDL, IDL and HDL) fractions of human and (for comparison) minipig blood plasma, which has not been studied yet. In order to obtain more accurate results, a new HPLC method based on the use of newly synthetized internal standards was developed. Four homologues of fenofibric acid prepared have identical chromophoric part of their molecules and hence the same UV spectra as fenofibric acid. From this point of view, these standards are more suitable for determination of fenofibric acid than the formerly used ones--naproxen or bezafibrate. Fenofibric acid levels in the high density lipoprotein fraction has been shown to be significantly higher (in both human and minipig plasma) than in the other lipoprotein fractions. This fact may be explained by higher affinity of the fenofibric acid to proteins constituting major part of the high density lipoprotein fraction. PMID- 9725496 TI - Evaluation of a bayesian pharmacokinetic program for phenytoin concentration predictions in outpatient population. AB - The present work evaluates the performances of a Bayesian program (PKS) for phenytoin concentration predictions in an outpatient population. The retrospective study involved 19 epileptic adults receiving oral phenytoin. The program was used to predict estimated serum concentrations from 0, 1, 2 or 3 feedback concentrations. Measurements of prediction bias (ME) decreased as soon as one steady-state concentration (Css) was used for estimations. Precision (MAE) was significantly improved with 1 Css and was even better and stable with 2 and 3 Css. Likewise, RMSE (composite of bias and precision) regularly decreased when the number of Css used increased. On a clinical way, 12% of the estimations were unacceptable (prediction error > 5 mg/l) with 1 Css and less than 3% with 2 or 3 Css. This number of rejected estimations increased to 45% when no feedback concentration was used. Besides, the program was able to predict important rises of serum levels in spite of relative low increase of the dose when 1 Css at least was known. Thus, the phenytoin dosing program has acceptable performances when at least 1 Css is known, and represents a potential tool to assist the clinician in the particular condition of outpatient population. PMID- 9725497 TI - OKT3 monitoring in the treatment of steroid-resistant acute rejection of hepatotransplant recipients. AB - OKT3 is a monoclonal antibody used as T-specific immunosuppressor agent in the treatment of acute rejection of hepato- or renal-transplanted patients. The immunosuppressor effect is related to the elimination and modulation of T-cells after the binding between OKT3 and the specific antigen CD3+. This drug has been used in the treatment of acute rejection. The more frequent side effects is the immunogenic reaction Human Antibody Mouse Antibody (HAMA). The aim of this study is the evaluation of the dose and the administration route of the OKT3. The results of the antibody monitoring in the plasma of the treated patients and the analysis of the clinical data were evaluated to focus a valid therapeutic protocol as well as a more rational time sampling of the circulating drug to achieve a correct monitoring. The results show a gradual increase of the hematic concentration of the drug, positively correlating the clinical data of hepatic biopsy and lymphocytic screening. These results have permitted to modify the therapeutic protocol previously performed. It has been defined the administration route choosing i.v. infusion (5 mg/die/2 h), moreover it the therapy has been shortened to 6 days. The HAMA were also evaluated and the analysis of the data showed a negative results, suggesting the possibility of the OKT3 retreatment in the cases of rescue. PMID- 9725498 TI - Surfactant effects on the in vitro percutaneous absorption of diclofenac sodium. AB - Nonionic surfactants, which are a safe class of enhancers, may offer means of enhancing drug permeation through the skin. In order to determine this effect, the influence of four nonionic surfactants on the percutaneous absorption of diclofenac sodium from carbopol gels containing 40% propylene glycol was investigated. In vitro diffusion experiments were carried out using excised full thickness abdominal rat skin as well as cellulose nitrate membranes. The data of this study clearly revealed that Tween 80 decreased diclofenac penetration rate. This was due to a decrease in thermodynamic activity as a result of micellar complexation. In contrast, the more hydrophobic sorbitans enhanced diclofenac skin penetration, probably due to changes in the barrier properties of the skin and in the vehicle-stratum corneum partition coefficient. The most enhancing effect was induced by Span 20, a surfactant with a C12 saturated hydrophobic group. However, diffusional lag times for all the tested surfactants were longer than for the control gel. PMID- 9725499 TI - An improved everted gut sac as a simple and accurate technique to measure paracellular transport across the small intestine. AB - An improved everted gut sac system has been developed in which the sacs were carefully prepared from rat small intestine and incubated in tissue culture medium. Under these conditions, the tissue showed good morphology at the electron microscope level, and was metabolically active for up to 2 h at 37 degrees C. Mannitol, an established probe of paracellular transport, was transported from the mucosal to the serosal side of the sac tissue. Excellent kinetic data showed that transport was linear up to 75 min and over a wide range of concentrations (0.025 - (10 mM). Mannitol was not detected in the tissue and transport was enhanced by EGTA, confirming the paracellular route of passage. Sacs prepared from colon also showed mannitol transport, but at a slower rate. Comparisons with Caco-2 cell monolayers showed that the everted sacs exhibited higher levels of paracellular transport than the cultured cell line. The improved everted gut sac system is an inexpensive and relatively simple technique with considerable potential as an in vitro tool to study the mechanisms, kinetics and enhancement of drug absorption across the small intestine at different sites and in the colon. PMID- 9725500 TI - Chromatographic investigation and computer simulation of L-deprenyl metabolism. PMID- 9725502 TI - Acetaminophen presystemic biotransformation vs bioavailability in therapeutical dosage range. AB - The influence of dose on the absorption and presystemic biotransformation of acetaminophen was studied in 15 healthy volunteers after administration of 3 different oral doses (250 mg, 500 mg, 750 mg) following a 3 x 3 Latin Square Design. The analytical method developed (High Performance Liquid Chromatography, HPLC) allows the rapid and simultaneous determination of acetaminophen and its major metabolites (its glucuronide and sulphate conjugates) by direct injection of urine samples. The statistical analysis did not reveal significant differences (alpha < 0.05) among treatments in the percentage of dose excreted and MRT and VRT values. Consequently, our results indicate that the absorption and biotransformation of acetaminophen is not affected by the dose in the usual therapeutical range. PMID- 9725501 TI - Determination of lansoprazole in biological fluids and pharmaceutical dosage by HPLC. AB - A simple and rapid (extractionless) high-performance liquid chromatographic method with UV detection at 230 nm was developed for the determination of lansoprazole in biological fluids and pharmaceutical dosage. Niflumic acid was added as internal standard. The separation was performed at ambient temperature on a C18 Spherisorb column with acetonitrile + 0.1 M sodium acetate (40:60, v/v, pH 7) as mobile phase. The retention time was 5.2 min for niflumic acid and 6.7 min for lansoprazole. The detection limit was 20 ng/ml using a 100 microl loop. The method was successfully applied to a pharmacokinetic study of lansoprazole in humans. PMID- 9725503 TI - Use of the repeated cross-over design in assessing bioequivalence: (within and between subjects variability - Schuirmann Confidence Intervals estimation). AB - In 1992, the Division of Bioequivalence in the Office of Generic Drugs published a guide to Statistical procedures for bioequivalence studies using a standard two treatments cross-over design (1). This paper describes the application of the guidelines to a practical protocol and the recent Proc MIXED (SAS) will be shown to be much more convenient than the traditional Proc GLM for theoretical and practical reasons (correct estimation of residuals, analysis of the within subjects variation, direct calculation of the Schuirmann 90% Confidence Intervals). This new procedure was applied to a study protocol on riluzole (Rilutek) including a replicate design with the within-subject and between subject variances being estimated on Cmax and AUC biopharmaceutic parameters. PMID- 9725504 TI - Biological and clinical study kinetics of biodegrading apatite-collagen implant for substituting bone tissue defects. AB - We studied kinetics of biotransformation synthetic implant for substituting defects of the bone tissue using methods of computer tomography, scintigraphy and morphology in vivo and in vitro. We found that the character of biotransformation is dependent on the degree of loaded or unloaded bone tissue. PMID- 9725505 TI - Treatment of lactose intolerance with exogenous beta-D-galactosidase in pellet form. AB - The effectiveness of a new beta-D-galactosidase pellet formulation in the treatment of lactose intolerance was studied. The encapsuled beta-D-galactosidase (lactase) pellets were first tested in vitro for their enzymatic activity within an environment simulating gastric conditions and subsequently within an environment simulating duodenal conditions. Effectiveness was measured by the % of glucose formed by hydrolysis of lactose. The pellets were found to retain their enzymatic activity in gastric pH conditions (mean 69 +/- 1 mg/dl glucose) and were found to hydrolyse lactose in human duodenal fluid (106.35 +/- 1 mg/dl). Finally the effectiveness of the new lactase formulation on glucose absorption was studied in 8 lactose intolerant subjects in a randomized, double blind, crossover trial. After fasting, the subjects were given one capsule containing 100 u/ml beta-galactosidase (i.e. 10 pellets of 10 u/ml each) or one capsule containing placebo pellets, followed by 100 g lactose dissolved in water. The washout period between lactose challenges was one week. Plasma glucose concentrations were measured before and at intervals after the challenges and the subjects completed symptom questionnaires every eight hours for 24 hours. Results showed a statistically significant increase in plasma glucose levels 30, 60, 90 and 120 min after lactose ingestion (repeated measures analysis of variance, p<0.01). Subjective ratings of the severity of abdominal cramping, belching, flatulence, vomiting and diarrhoea were significantly decreased following ingestion of the lactase pellets and lactose (no incidence of diarrhoea) compared with after ingestion of placebo and lactose. The results of the study were considered to be very promising as the beta-D-galactosidase formulation (which was produced at very low cost and with great ease) resisted inactivation in the stomach, effectively transformed lactose to glucose in vivo and reduced symptoms of lactose intolerance. PMID- 9725506 TI - Ethics and the progress of medical sciences. PMID- 9725508 TI - The vascular center at Brigham and Women's Hospital. PMID- 9725507 TI - Vascular Center Program: historical perspective and Northwestern experience. PMID- 9725509 TI - The Mayo Vascular Center experience. AB - American medicine is trending toward an increasing number of specialty care centers. Cancer centers, transplant centers, and sports medicine centers are only a few common examples. Vascular centers are relatively new entities that are forming for obvious reasons. As the general population ages, peripheral vascular disease has become more prevalent. Several types of medical, surgical, and radiological specialists are involved in the diagnosis and treatment of such patients. Creating multispecialty vascular centers is one method to focus expert care on the patient, to alleviate some of the turf battles between specialties, and to contain burgeoning Medicare costs. PMID- 9725510 TI - Unique aspects of anticipated shipboard vascular trauma. AB - This article mentions notable historical examples of vascular injuries that occur at sea. It traces the development of modern ships and the concomitant capability to provide medical care to the personnel who go in harm's way on these ships. The importance of vascular surgery training for the general surgeon assigned to sea duty is stressed. PMID- 9725511 TI - Surgical approach to military vascular injuries. AB - PURPOSE OF THIS STUDY: Vascular injuries caused by high-velocity military missiles are associated with bone fracture, soft-tissue, nerve and tendon injuries. In this study we will discuss the surgical strategy and results of vascular injuries, which require a different approach from primary and elective surgical procedure. BASIC METHODS: Surgical interventions were performed in 116 patients. Vascular lesions were localized on the lower extremity in 53, upper extremity in 55, and nine were in other regions. Vascular injuries were concomitant with bone fracture in 46 and nerve injuries in 36 patients. Vascular repair was performed after orthopedic stabilization in vessels with an ischemic period of less than 4 hours. PRINCIPAL FINDINGS: Fasciotomy was performed after vascular repair in the 22 cases that had arrived after 8 hours. Amputation was required in two cases. There was one mortality. CONCLUSIONS: The best results are obtained when a multidisciplinary and emergency approach are used by the team of vascular, orthopedic, plastic and neurosurgeons who are experienced in military injuries. PMID- 9725512 TI - Cryptogenic Salmonella-infected ruptured aortic aneurysms. AB - Salmonella-infected ruptured aortic aneurysms are a difficult surgical problem. Five patients with Salmonella-infected ruptured aortic aneurysms are presented. All patients were initially treated by in situ graft replacement, wide debridement of the infected aortic tissue and long-term systemic antibiotic therapy. The surgical technique of choice in acute situations, such as a rupture of the aneurysm, remains controversial. In the recent literature 26 reported cases were reviewed. Based on our clinical experience and these case reports, in situ bypass reconstruction can be justified as a surgical technique that gives at least a good short-term prognosis. PMID- 9725513 TI - Infrainguinal reconstructions: influence of surgical experience on outcome. AB - OBJECTIVES: To evaluate the influence of surgical experience and operation technique on the outcome of infrainguinal reconstructions. DESIGN: A longitudinal observational study of patients undergoing reconstructions to the popliteal and crural arteries. SETTING: A regional hospital and an academic referral centre. MATERIALS: 392 patients undergoing 442 reconstructions, 218 to the popliteal and 228 to the crural vessels. CHIEF OUTCOME MEASURES: Graft patency and leg salvage. MAIN RESULTS: At the popliteal level, 5-year patency after autogenous vein reconstruction was 76%. No difference was found between in situ and transposed vein reconstructions, but prosthetic reconstructions had a worse patency. At the crural level, 5-year patency for in situ vein, transposed vein and prosthetic bypass were 66, 55 and 21%, respectively. Corresponding leg salvage for chronic critical leg ischaemia was 91% with vein and 55% with prosthetic reconstruction at popliteal level, and 66 and 33% at the crural level. Surgical experience improved the outcome, as 3-year patency for in situ and transposed vein bypasses to the popliteal level were 82 and 95% for experienced surgeons, and 53 and 75% for less experienced surgeons. Corresponding leg salvage rates for reconstructions to the popliteal and crural levels were 85 and 67% for experienced surgeons, and 61 and 52% for less experienced surgeons. CONCLUSIONS: Outcome of infrainguinal reconstructions is influenced by the reconstruction level. Surgical experience and choice of the appropriate reconstruction technique can improve outcome. PMID- 9725514 TI - Diagnosis of limbs and neck arterial trauma using duplex ultrasonography. AB - OBJECTIVE: To evaluate duplex ultrasonography for diagnosis of arterial trauma in limbs and neck. METHOD: Fifty-one wounds in 47 patients, with indication for arteriography, were prospectively studied and grouped according to the presence (PCS group: 21 wounds, 41.2%) or absence (ACS group: 30 wounds, 58.8%) of clinical signs of arterial injury. All underwent duplex ultrasonography and arteriography. RESULTS: Arteriography disclosed arterial injury in 21 wounds, of which 19 were visualized by duplex ultrasonography. In the other 30 wounds neither methods disclosed any arterial injury. The sensitivity of duplex ultrasonography was 90.5%, the specificity was 100% and the accuracy was 96.1%. In PCS group duplex ultrasonography showed 14 injuries (93.3%) and one false negative result, and in ACS group, five injuries (83.3%) and one false-negative result in the ACS group. CONCLUSIONS: Duplex ultrasonography reproduces the results of arteriography as a non-invasive diagnostic method in trauma of the limbs and neck. PMID- 9725515 TI - Endothelial damage due to ischemia and reperfusion is prevented with SIN-1. AB - BACKGROUND: Acute ischemia followed by reperfusion results in direct endothelial damage characterized by cell swelling, increased permeability and loss of acetylcholine-mediated vasorelaxation. Ischemia followed by reperfusion in a New Zealand white rabbit hindlimb has been shown to result in loss of acetylcholine induced relaxation of superficial femoral arteries. This loss of relaxation in response to acetylcholine is a reflection of the decreased nitric oxide availability that occurs with reperfusion injury. The purpose of this investigation was to evaluate the effect of SIN-1, a direct nitric oxide donor, on this endothelial injury. METHODS: New Zealand white rabbits underwent complete ischemia of the right hindlimb for 3 h followed by 2 h of reperfusion. Aliquots of 20 ml of either 0.88-mM SIN-1 or normal saline was infused via a lateral branch of the right common iliac artery during the first 20 min of reperfusion. Sham vessels were subjected to the 5-h operative intervention to control for anesthetic effect. Control vessels were harvested from rabbits not exposed to ischemia or reperfusion. Superficial femoral artery rings were evaluated in vitro for endothelial cell-mediated relaxation. Rings were contracted with potassium chloride and norepinephrine and then exposed to standardized incremental doses of acetylcholine to measure percent relaxation. Artery sections were sent for hematoxylin and eosin staining. RESULTS: No significant differences were seen in contraction caused by either potassium chloride or norepinephrine in all four experimental groups. Saline infused vessel rings relaxed a mean of 8.42 +/- 2.39% and 49.57 +/- 8.65% in response to acetylcholine doses of 3 x 10(-8) M and 1 x 10(-7) M, respectively. In contrast, SIN-1 infused vessels relaxed a mean of 57.82 +/- 2.65% and 100.23 +/- 1.53% to the same doses of acetylcholine. Control and sham arteries showed a similar relaxation response as compared with SIN-1 infused vessels. Differences in relaxation when comparing saline infused vessels with SIN-1 infused, sham and control arteries, were significantly different at each dose of acetylcholine from 3 x 10(-8) M to 1 x 10(-7) M (P < 0.05, ANOVA). Histologic examination of the vessels revealed no morphologic differences among the experimental groups. All vessels were structurally normal with an intact endothelium. CONCLUSION: In this model of rabbit hindlimb ischemia, preservation of endothelial cell-mediated vasorelaxation occurs with administration of intra arterial SIN-1 during reperfusion. This preservation of endothelial function cannot be explained by histologic changes in the arterial wall or attributed to altered arterial contractility in response to potassium chloride or norepinephrine. PMID- 9725516 TI - Transcranial cerebral oximetry during carotid endarterectomy: agreement between frontal and lateral probe measurements as compared with an electroencephalogram. AB - OBJECTIVE: Transcranial cerebral oximetry, which is considered a novel technique, was evaluated during carotid endarterectomy. For practical reasons, the use of a single probe attached to the forehead and overlying the territory of the anterior cerebral artery is recommended. Other monitoring systems (transcranial Doppler, electroencephalograms (EEG)) focus more on the territory of the middle cerebral artery. The aim of this study was to evaluate whether a probe in the frontal area is as representative for monitoring cerebral ischaemia during carotid cross clamping as a probe in the lateral area. DESIGN: Clinical prospective study. MATERIALS: Sixty patients who underwent carotid endarterectomy were studied with continuous and simultaneous EEG and transcranial cerebral oximetry. Forty-three patients (72%) simultaneously underwent frontal and lateral transcranial cerebral oximetry. The lateral probe was only used in 17 patients. METHODS: The percentage decrease of transcranial cerebral oximetry was calculated during cross-clamping. Using the EEG findings as the gold standard in order to detect cerebral ischaemia during carotid cross-clamping, the relationship with transcranial cerebral oximetry was described in terms of sensitivity, specificity and the area under the curve in a receiver operating characteristic curve. RESULTS: The 95% confidence interval of the area under the curve of the receiver operating characteristic of the lateral probe was 0.61-1.00 and that of the frontal probe was 0.65-1.00; therefore there is virtually no difference between the two methods. In 18% of the cases the lateral transcranial cerebral oximetry was hindered by practical failures. CONCLUSION: Considering the lack of additional information and the practical problems experienced with the lateral probe, it was concluded that transcranial cerebral oximetry with a single frontal probe is a practical non-invasive monitoring system and is at least as representative for monitoring cerebral ischaemia during carotid cross-clamping as a lateral probe. PMID- 9725517 TI - Experience with cryopreserved arterial allografts in the treatment of prosthetic graft infections. AB - The authors present a retrospective study on 30 patients with prosthetic graft infection. Included are 25 patients with aortic graft infection, three with infection of a femorodistal bypass and two with infected axillofemoral grafts. There were 23 isolated primary prosthetic graft infections and seven aorto enteric fistulas. Treatment consisted of graft excision and replacement with cryopreserved arterial homografts, harvested from brain-death multi-organ donors. The in situ technique was used in 27 cases. Eight patients died postoperatively and two deaths were from allograft related complications. The operative mortality rate was 11% for isolated aortic graft sepsis and the early limb salvage rate was 100%. Persistent or recurrent infection was noted in two cases. The mean follow up of the series was 24.5 months and occlusive complications occurred in five patients (23%), which resulted in two major amputations. Serial CT scans showed abnormalities in six of the 22 survivors, all of them related to the aortic segment of the allograft. It is concluded that in situ reconstruction with cryopreserved arterial allografts represents an acceptable alternative, especially in the treatment of isolated aortic graft sepsis. Continued follow-up towards late deterioration and/or occlusive complications remains mandatory. PMID- 9725518 TI - Prevention of limb loss in critical ischaemia by arterialization of the superficial venous system: an experimental study in dogs. AB - The possibility of ischaemic limb salvage by arterializing the superficial venous system was examined in a canine study. The experiment was carried out on four healthy dogs in three stages. In the first stage, collateral circulation to the hind limb was abolished. In the second stage, all branches of the common femoral artery were ligated, which created a model of ischaemia incompatible with limb survival. Revascularization was achieved by anastomosing the valvulotomized long saphenous vein to the common femoral artery, proximal to its ligation. The dogs were monitored for 2 weeks. All limbs maintained tissue oxygenation similar to that of the control contralateral limb. In the third stage, the artery was ligated proximal to the femoro-saphenous anastomosis and the limb monitored for 7 hours. Acute loss of motor function of the limb resulted. In the present study, arterialization of the valvulotomized long saphenous vein prevented limb loss in critical ischaemia. PMID- 9725519 TI - Sequential vein bypass grafting: tactics and long-term results. AB - The sequential bypass grafting technique has many advantages over coronary artery bypass grafting with single grafts. The aim of this study was to evaluate the consequences of sequential bypass graft failure. Between 1 January 1984 and 31 December 1996, 3846 patients underwent primary coronary artery bypass vein grafting. A total of 3490 patients received sequential vein bypass grafts and 356 patients received single vein bypass grafts (9%). There were 6177 sequential bypass grafts (3490 postero-lateral grafts (56%) and 2687 in the antero-lateral position (44%)) and 1468 single grafts (972 vein grafts and 496 internal thoracic artery grafts). Overall, there were 80 hospital deaths (2.1%). Mortality in relation to type of grafts used was: 13 deaths in 356 patients with only single graft (3.7%) and 67 deaths in 3490 patients who received sequential vein grafts (1.9%). Of 3766 hospital survivors, 3731 were followed for an average of 76 months. During follow-up, 85 patients died (2.3%), 15 patients (0.4%) underwent cardiac transplantation and 52 (1.4%) had re-do coronary artery bypass vein grafting. Graft-percutaneous transluminal coronary angioplasty was performed in 56 patients (1.5%), 37/1390 single bypass grafts (2.7%) and 19/6023 sequential bypass grafts (0.3%). There were 272/6023 symptomatic sequential graft occlusions (4.5%) (182 were in postero-lateral position and 90 in the antero-lateral position). There were 66/667 single vein graft occlusions (9.9%) and 15 symptomatic internal thoracic artery graft occlusions (2.1%) during follow-up. In 97% of patients, presenting symptoms of postero-lateral sequential bypass graft occlusion took the form of a renewed angina with a myocardial infarction rate of 3% and a mortality rate of 7%. Corresponding figures for antero-lateral sequential bypass grafts were 22, 78 and 68%, and anterior single vein bypass grafts were 70, 30 and 15%, respectively. The overall 10-year survival rate in patients with sequential bypass grafts was 81.2% and the cumulative patency rate (1464 angio-controls of 2576 sequential vein grafts) was 72.2%. A symptomatic occlusion of a postero-lateral sequential vein bypass results in a low incidence of myocardial infarction with low mortality, when the terminal anastomosis is connected to a high flow vessel. An antero-lateral sequential vein bypass graft has better long-term patency than single vein bypass, but should occlusion occur, it would usually be associated with a higher myocardial infarction and mortality rates than a single vein graft. The highest risk for failure of a sequential graft in the antero-lateral position occurs when the left anterior descending artery (LAD) is small or severely diseased. In this situation the single graft technique with internal thoracic artery appears to be safer. PMID- 9725520 TI - Moderately hypothermic cardiopulmonary bypass and selective cerebral perfusion in ascending aorta and aortic arch surgery. Preliminary experience in twenty-two patients. AB - Deep hypothermic cardiocirculatory arrest is the commonest method of brain protection during transverse aortic arch surgery. Its principle drawbacks consist in the limited safe ischemic period and in the coagulative, renal and pulmonary complications related to low body temperatures and prolonged cardiopulmonary bypass time. Different selective cerebral perfusion techniques have recently raised the interest of some surgical teams in an effort to obviate these problems. The authors' initial experiences with 22 patients, ranging in age from 19 to 78 years (mean, 55 +/- 15 years), who underwent ascending aorta and/or aortic arch replacement using selective cerebral perfusion and moderately hypothermic cardiopulmonary bypass are reported here. Acute aortic dissection and atherosclerotic aneurysm were the commonest lesions observed: ascending aorta associated with partial or complete arch replacement was the most widely performed procedure. With regard to the perfusion technique, after regular cardiopulmonary bypass had been established through the iliac vessels, selective cerebral perfusion was started after aortic arch vessels cannulation (innominate artery, bilateral common carotid artery, innominate artery and left common carotid artery, or right common carotid artery) using a single roller pump separately from the systemic circulation, and brain perfusion was achieved by blood cooled at 30 degrees C, at a flow rate that ranged from 300 ml/min to 1500 ml/min, at a perfusion pressure of approximately 65 mmHg, with the patient maintained at moderate hypothermia (30 degrees C rectal). To perform distal aortic repair, if transverse aortic arch or proximal descending aorta cross clamping was not feasible, cardiopulmonary bypass flow was lowered to 300-350 ml/min and an open anastomosis was performed, while independently assuring cerebral perfusion (six patients). There were three hospital deaths (mortality rate of 13.6%; s.d. 6.0-25.5%; 70% confidence limit), but none because of cerebral accident. No paraplegia occurred. One patient suffered from right hemiparesis, neither renal nor pulmonary complications were observed. Two chest reexplorations were necessary for bleeding, which were partially related to hemocoagulative disorders. In our experience, the technique of moderately hypothermic cardiopulmonary bypass and selective cerebral perfusion in aortic surgery has provided good results with regard to cerebral protection and organ function preservation. Therefore, allowing a prolonged distal aortic reconstruction period, it may be considered as a safe alternative to profound hypothermia associated with cardiocirculatory arrest in aortic arch surgery. PMID- 9725521 TI - Inflammatory response and oligo-element alterations following cardiopulmonary bypass in patients undergoing coronary artery bypass grafting. AB - Oligo-elements such as zinc (Zn), selenium (Se) and copper (Cu) have a significant influence on the function of the immune system. Various immunological and inflammatory changes are known to occur in patients undergoing cardiopulmonary bypass. The aim of this study was to evaluate changes in serum oligo-elements levels during and following cardiopulmonary bypass. The serum levels of Zn, Se and Cu were determined in 67 consecutive patients, with coronary artery disease admitted for coronary artery bypass grafting. Blood samples for oligo-elements, analysis were withdrawn into metal-free tubes just prior to the start of cardiopulmonary bypass; at 30, 60 and 90 min into cardiopulmonary bypass; following weaning from cardiopulmonary bypass; 30 min after termination of cardiopulmonary bypass; at 24 h; and on the 5th postoperative day. Trace elements analyses were performed using atomic absorption spectrophotometry. Interleukin 6 and 8, as well as serum albumin, creatine phosphokinase, lactate dehydrogenase and creatine phosphokinase-MB fractions were also analyzed. The mean age was 63 +/- 9 years and 91% (61) were men. The mean preoperative left ventricular function was 52 +/- 12%, Canadian Cardiovascular Society (CCS) angina class was 3.7 +/- 0.5 and 30% (20) of the operations were re-do's. All patients had normothermic cardiopulmonary bypass. Mean cardiopulmonary bypass-time was 85 +/- 31 min. One patient was lost for the recovery sampling (hospital mortality, 1.5%). Nine patients had a postoperative cardiac index < 2.0 liter/min per m2, which required pharmacological support and additional intra-aortic balloon pump in two of them. Other postoperative complications were few. There was a rapid depletion of S-selenium and S-Zn levels, which were halved at 30 min after cardiopulmonary bypass and remained low throughout the study period. The Cu/Zn ratio increased significantly at the start of cardiopulmonary bypass, which indicated an inflammatory reaction and was not normalized until the 5th postoperative day. Length of ischemia time, presence of diabetes. hypertension and hyperlipidemia did not influence the results, while a prolonged cardiopulmonary bypass-time > 120 min resulted in a higher Cu/Zn ratio than observed for shorter cardiopulmonary bypass-times. This indicates a more profound inflammatory response. Inflammatory parameters responded in the same manner as described earlier by others. These data indicate that severe loss of various oligo elements occur in patients undergoing coronary artery bypass grafting and suggests that a supplementary administration of zinc and perhaps also selenium could be appropriate during cardiopulmonary bypass. PMID- 9725522 TI - Radical debridement and omental transposition for post sternotomy mediastinitis. AB - OBJECTIVE: Reported mortality for postoperative mediastinitis treated by debridement alone can reach 40%. The authors' experience with radical debridement and omental transposition is reviewed. METHODS: Between May 1990 and August 1996, 14 patients with untractable mediastinitis had a transfer of the greater omentum: 11 after coronary artery bypass grafting (CABG) (6 bilateral internal thoracic arteries ITA grafts), one after a heart transplant, one after an aortic valve replacement and CABG, and one after a repair of the aortic isthmus related to a motor vehicle accident. The mean age was 63 +/- 8 years. Infection was proven in all patients by cultures of intraoperative specimens. Two patients had such a large sternal defect that no primary closure could be attempted. The remaining 12 patients had a mean of 1.4 +/- 0.7 previous debridement. Five patients had a total sternectomy. After radical debridement, the omentum was transferred over the entirety of the wound and covered with a meshed thin skin graft. All patients had a minimum of 4 weeks of i.v. antibiotic therapy. RESULTS: There was no operative death. Apart from one focal necrosis and one traumatic dehiscence of the omentum, there was no hospital complication. Sepsis was controlled in all patients. The median hospital stay was 31 days (range 20-154 days). At a median follow-up of 20 months (range: 6-44 months), there were two late deaths: one sudden and unexpected death and one after a re-do CABG. The remaining patients had resumed their previous activities. One patient had developed an incisional hernia and another underwent further surgery for cosmetic reasons. CONCLUSION: Radical debridement and omental transposition may achieve a cure for postoperative mediastinitis with good mid-term results. PMID- 9725524 TI - Critical lower limb ischaemia following excision of the rectum. AB - Critical lower limb ischaemia can occur following rectal surgery by a number of mechanisms. Patients with aorto-iliac stenosis or occlusion may be dependent on collateral circulation to the lower limbs from the visceral arteries supplying the descending colon, sigmoid colon and the rectum. Division of these collaterals can precipitate critical ischaemia of the leg. This is an uncommon scenario but one that should be considered in arteriopaths undergoing rectal surgery. Two cases of this complication are reported and the mechanisms discussed. PMID- 9725523 TI - Use of the right gastroepiploic artery as a coronary artery bypass graft in 307 patients. AB - From October 1988 to October 1995 the right gastroepiploic artery was used as a conduit for coronary surgery in 307 patients. Their average age was 56.5 years (range 25-75) and 274 patients (89%) were male. Twenty-six cases (8.5%) were re operations and 58 patients (19%) were operated upon on an urgent or semi-urgent basis. Target coronary vessels were the right coronary artery and its branches in 280 cases (91.4%), the circumflex artery in 25 cases (8%) and the left anterior descending artery in two cases. The right gastroepiploic artery was used as an in situ graft in 303 cases (98.7%) and as a free graft in 4 (1.3%). A total of 291 patients (94.8%) also received at least one mammary artery graft: both mammary arteries were used in 167 patients (54.4%). An average of 3.6 distal anastomoses were made per patient, three of them with arterial grafts. Eleven (3.2%) right gastroepiploic artery grafts were doubled with saphenous vein intraoperatively because of persistent myocardial ischemia. In-hospital mortality was 1.6% (five patients). Perioperative myocardial infarction occurred in twelve patients (3.9%). Follow-up now averages 26 months (range 6-88). There have been five late deaths (1.6%). A total of 265 (89.2%) patients are angina free. Of the total, 145 patients have been investigated with a maximal-stress test coupled with scintigraphy: residual myocardial ischemia was found in 10 patients, right gastroepiploic artery was related in three. Ninety-six patients have undergone angiographic restudy at a mean of 12 months (range 8-88) postoperatively. Patency of the right gastroepiploic artery grafts was 91.8%. This study confirms that the right gastroepiploic artery can be used as a conduit for coronary artery bypass surgery with minimal mortality or morbidity. Mid-term patency rates and clinical outcome are encouraging. PMID- 9725525 TI - AIDS in ENT in children. AB - Human immunodeficiency virus (HIV) infection is one of the most widespread diseases in the world. By the end of 1995, 800,000 HIV infected persons were suspected in Thailand, although the reported number of symptomatic HIV patients was only 13,267 and the number of cases of acquired immunodeficiency syndrome (AIDS) was 31,439. Approximately 5.2% of AIDS patients are cases of paediatric AIDS, contracted mostly by perinatal transmission and with a 25% vertical transmission rate. In a study of paediatric AIDS patients in the Children's Hospital, Thailand, from 1992 to 1995, the five most common clinical manifestations were hepatosplenomegaly (82.85%), persistent pneumonia (64.4%), oral candidiasis (59.6%), chronic diarrhoea (58.4%) and failure to thrive (51.2%). In addition to oral candidiasis, other ENT (ear nose-throat) presentations were lymphadenopathy (41.6%), repeated upper respiratory tract infection (39.5%), otitis media (18.4%), parotitis (5.2%) and sinusitis (0.8%). PMID- 9725526 TI - Acquired tracheo-esophageal fistula in the pediatric population. AB - Acquired tracheo-esophageal fistula (TEF) in the paediatric population is a rare entity, an acquired fistula can be due to tracheotomy tubes and tracheotomy cuffs. Patients with burns are at greater risk from these tracheotomy complications. Acquired TEF can also occur due to a foreign body impaction. Prevention and early diagnosis are important. In patients with possible airway burns, 'safe' intracuff pressures maybe too high. To avoid further damage of the mucosa, the patient should have a small air leak maintained if a cuff is used. Most acquired TEF do not close spontaneously and surgical closure is required. Our paper presents two cases of acquired TEF in the pediatric population and reviews the literature on this subject. PMID- 9725527 TI - The development of a paediatric quality of life questionnaire to measure post operative pain following tonsillectomy. AB - In this paper we describe the process through which a Paediatric Quality of Life Questionnaire was developed to assess the pain experienced by children following tonsillectomy. The impetus behind the questionnaire development was the clinical observation that the pain experienced by children undergoing tonsillectomy was not being detected and analgesia was not being prescribed in adequate amounts. The eight-item questionnaire provides the independent observer with an objective measure of the physical and emotional distress a child experiences post tonsillectomy to which analgesic requirements can be titrated. The questionnaire was tested on a sample of 48 children aged between 2 and 13 years (average age 7.1 years) who underwent elective dissection tonsillectomy by the same surgeon using a standardised operative technique. The conclusions substantiate the hypothesis that there is the need for an assessment tool based on non-verbal behaviour to measure post-operative pain in children. In the discussion we summarise the increasing role of quality of life assessments in surgical practice. PMID- 9725528 TI - Effect of contralateral masking on the latency of otoacoustic emissions elicited by acoustic distortion products. AB - Otoacoustic emissions (OAE) are sound products generated by the outer hair cells (OHC) in the inner ear. The OHC are capable of moving spontaneously or in response to acoustic stimuli (spontaneous otoacoustic emissions and evoked otoacoustic emissions), these movements are known as electromotility. Electromotility is affected when contralateral acoustic stimulation is introduced to the ear. Different types of stimuli may produce this response. Clicks, pure tones, and white masking noise have been used as contralateral stimulation. This effect appears to be mediated by the medial efferent olivocochlear bundle. Contralateral masking produces suppression of OAE, especially on the amplitude. However, the effect of contralateral masking on the latency of distortion product otoacoustic emissions (DPOAE) has not been studied. The purpose of this paper is to investigate whether contralateral masking, with wide band masking noise, may produce a significant change on the latency of the DPOAE. Three different latency measurements of DPOAE measurements were made on low, middle and high frequencies of fl including 574 Hz, 2454 Hz and 4919 Hz. Each one of these frequencies was measured with and without contralateral masking. Twenty-eight ears of 15 subjects were studied. Non-significant differences (P > 0.05) between masked and unmasked conditions were found in all cases. It is concluded that contralateral masking does not appear to affect latency of DPOAE. PMID- 9725529 TI - Temporomandibular joint dysfunction in children: evaluation of treatment. AB - Thirty-eight pediatric patients with temporomandibular joint (TMJ) dysfunction were diagnosed and treated. The etiology for the TMJ dysfunction was trauma in 30 (79%) patients, degenerative joint disease in two (5%) patients, growth disturbances in two (5%) patients and tumor in one (3%) patient. In three (8%) patients the etiology was unclear. The treatment modalities were: non-invasive therapy in 19 (50%) patients, occlusal therapy in 10 (26%) patients and surgical treatment in nine (24%) patients. The reported symptoms of temporomandibular joint dysfunction using the Helkino anamnestic index revealed that, at the initial examination, eight (21%) reported mild symptoms and 30 (79%) severe symptoms. One year later, 33 (87%) reported no symptoms, four (10%) mild symptoms and one (3%) severe symptoms. These differences were significant (P < 0.05-0.01). Maximum mouth opening 1 year after treatment as compared to the initial examination increased (P < 0.05) in all three treatment modalities. Deviation of the mandible on opening, 1 year after treatment as compared to the initial examination, decreased (P < 0.05) in all three treatment modalities. No differences were found between the modalities in both the maximum mouth opening or deviation of the mandible. TMJ dysfunction in children can be effectively treated by non-surgical treatment modalities. Surgery should be considered only when the non-surgical therapies were ineffective. PMID- 9725530 TI - Otologic management in children with the CHARGE association. AB - OBJECTIVES: To characterize otologic management of two patient groups, those with the CHARGE association and those not strictly labeled as CHARGE but with several features of the disorder (CHARGE-like), in order to determine: (1) the clinical validity and utility of managing CHARGE-like children in a similar manner to patients with the strictly defined CHARGE association, (2) the progression and prognosis of hearing loss and (3) the identification of factors that may predict the degree of hearing loss. DESIGN: Case series. SETTING: Tertiary care urban children's hospital. PATIENTS: 37 children, 22 in the CHARGE group and 15 in the CHARGE-like group. INTERVENTIONS: Otorhinolaryngologic and audiologic management. MAIN OUTCOME MEASURES: Otorhinolaryngologic and audiologic evaluation. RESULTS: All patients required otologic and/or audiologic care. Bilateral hearing loss was found in 32 patients (86%) and unilateral hearing loss in five patients (14%) when hearing was assessed in the absence of otitis media. Among the 32 patients with bilateral hearing loss, 31 (97%) were able to be fit with useful hearing aids. External ear anomalies were present in 25/37 (68%) patients, and middle ear and ossicular anomalies were identified in four cases (4/37, 11%), 36/37 (97%) patients required surgical management of otitis media. Three patients (3/37, 8%) exhibited radiographic evidence of inner ear deformity. Facial nerve dysfunction was noted in the records of 14/37 (38%) patients. No statistically significant difference was found when CHARGE and CHARGE-like patients were compared for degree of hearing loss (P = 0.5964), type of hearing loss (P = 0.2657), worsening of hearing level (P = 0.7908), or anomalies of the external ear (P = 0.6921), ossicles (P = 0.7908), inner ear (P = 0.7908) or facial nerve (P = 0.6409). Patients with external ear anomalies did not exhibit statistically different degrees (P = 0.3125) or types (P = 0.1515) of hearing loss from patients without auricular anomalies. The presence of facial nerve anomaly correlated significantly (P = 0.0021) with profound hearing loss. CONCLUSIONS: Children who are CHARGE-like may be may be considered equivalent in terms of otologic and audiologic management to children strictly defined as CHARGE patients. These children all require otologic care due to the high prevalence of middle ear disease and the underlying permanent hearing loss that is both stable and aidable. The degree of hearing loss cannot be predicted by external ear morphology, but may be predicted by facial nerve palsy. PMID- 9725531 TI - Safety of powered instrumentation for adenoidectomy. AB - A recent study established the utility of an endoscopic shaver for adenoidectomy in children by the transoral approach and showed that power assisted adenoidectomy (PAA) was significantly faster with a trend toward decreased blood loss. The purpose of this study was to demonstrate the safety of power assisted adenoidectomy in a large cohort of patients. A retrospective review was performed of 329 patients who had adenoidectomy by powered instrumentation. Postoperative complications were documented and compared with a similar group that had curette adenoidectomy. Complications watched for included prolonged recovery, postoperative hemorrhage, readmission for dehydration, velopharyngeal insufficiency, and nasopharyngeal stenosis. No postoperative complications were seen in the power assisted adenoidectomy group. This review confirms the safety of power assisted adenoidectomy. PMID- 9725532 TI - Nasal dermoid cysts in siblings. AB - Nasal dermoid cysts are congenital malformations which result from anomalous embryological development. Two cases occurring in siblings are presented. There have been several previous reports that the condition may be familial but in this report the initial suspicion of the milder anomaly in the younger child was raised primarily because of his older brother's history. This further report suggests that the incidence of familial occurrence of this anomaly may be greater than current literature suggests, and the possibility that other family members may be affected should be remembered by clinicians treating patients with this condition. PMID- 9725533 TI - Supernumerary nostril: a rare congenital deformity. AB - Duplication anomalies of the nose include polyhinia (double nose) and supernumerary nostril (assessory nostril). These are rare congenital nasal deformities resulting from aberrant embryological development. Differential diagnoses include glioma, encephalocele, nasal dermoid, nasolacrimal duct duplication, mid facial cleft and proboscis lateralis (K. Nakamura, T. Onizuka. Plast. Reconstr. Surg. 80 (3) (1987) 436-441). Our review of the English language literature revealed eight reported cases of duplication anomalies of the nose. Four of these were cases of polyrhinia (double nose). Of the cases remaining, one patient had a supernumerary nostril in association with a cleft lip, leaving only three reported cases of an isolated supernumerary nostril. We present a newborn infant with an isolated right supernumerary nostril. MRI, CT and photographic documentation are provided. Pertinent embryology, anatomy and a thorough review of the literature are included. PMID- 9725534 TI - Impacted aural foreign body requiring endaural incision and canal widening for removal. AB - Foreign bodies of the ear are a common finding in children. The majority of these can be removed relatively easily. We present a case which required endaural incision and widening of the external meatus before the foreign body could be successfully removed. Entrapment of the hard, oval-shaped, smooth and slippery foreign body deep to the hour-glass narrowing of the external canal, proximity of the object to the tympanic membrane and localised tissue oedema were the indications for the surgery. This case demonstrates the danger of unskillful attempts to remove aural foreign bodies. PMID- 9725535 TI - Sinus histiocytosis with massive lymphadenopathy (Rosai-Dorfman disease). AB - Sinus histiocytosis with massive lymphadenopathy (SHML) is a distinct clinicopathological entity described by Rosai and Dorfman and differentiated by other childhood histiocytoses by its distinct characteristics. This is a rare pathology and should be kept in mind for differential diagnosis of neck masses, especially in childhood. In this article a case with SHML is presented and clinical features of the disease given. PMID- 9725536 TI - Endoscopic adenoidectomy in a case of Scheie syndrome (MPS I S). AB - The mucopolysaccharidoses (MPS) are a heterogeneous group of relatively rare, progressive, inherited lysosomal storage disorders, characterized by a deficiency of lysosomal enzymes which are responsible for the stepwise degradation of glycosaminoglycans. Their deficiency leads to the accumulation of glycosaminoglycans in various organs causing progressive disruption of cellular functions and multiple systemic effects including otolaryngological problems, upper airway obstructive disease being the most common. Scheie syndrome (MPS I S) is due to the deficient activity of alpha-L-iduronidase leading to the intralysosomal accumulation of dermatan sulfate and heparan sulfate. We present our experience in one such case occuring in a 6-year-old girl with ENT manifestations and who underwent a successful endoscopic adenoidectomy for symptomatic adenoid hypertrophy. This procedure was preferred over a conventional adenoidectomy in order to avoid complications associated with abnormal cervical vertebrae. PMID- 9725537 TI - Internal jugular vein thrombosis in a child due to a 'pencil point injury' of the palate. AB - Penetrating trauma of the soft and hard palate are common in children and have been termed 'pencil point injuries.' Although such injuries are usually minor, the English literature has reported over 25 cases complicated by thrombosis of the internal carotid artery. We describe an unusual case of a 6 year old girl who presented with fever, cervical swelling and torticollis, following a pencil point injury. Physical examination and CT scan confirmed the diagnosis of internal jugular vein thrombosis (IJVT). The management of pencil point injuries and IJVT in children is reviewed and the possible mechanisms of IJVT in the case described here, are discussed. PMID- 9725538 TI - News from the Interamerican Association of Pediatric Otorhinolaryngology II IAPO's Congress, Sao Paulo, Brazil, September 1997 Presidential Address: challenges of emerging and reemerging diseases. PMID- 9725539 TI - News from the Interamerican Association of Pediatric Otorhinolaryngology II IAPO's Congress, Sao Paulo, Brazil, September 1997 Presidential Address: on pollution. PMID- 9725540 TI - The relevance of T-type calcium antagonists: a profile of mibefradil. AB - L- and T-type voltage-dependent transmembrane calcium channels are important for normal functioning of the cardiovascular system. T-type channels are a heterogeneous group, and have physiologic and pathophysiologic relevance in a number of organ systems, including the heart and central nervous system. They appear to be involved in the control of blood pressure in patients with essential hypertension and in protection from ischemic damage. Alterations of both L- and T type calcium channels are involved in the development of hypertension. Pharmacologic modulation of T-type calcium channels appears to reduce membrane calcium flux and ameliorate hypertension. During early ischemic damage, T-type calcium channels appear to remain functional whereas L-type channels are already inactivated. T-type calcium channels also appear to be involved in the development of supraventricular arrhythmias, some forms of arrhythmias in cardiomyopathy, and cardiac hypertrophy. The heterogeneity of T-type calcium channels should make it possible to target drugs to specific subgroups of T-type calcium channels. A new class of calcium antagonist, the benzimidazolyl substituted tetraline derivatives, has been shown to block both L- and T-type calcium channels. The first member of this class approved for clinical use is mibefradil. Clinical studies have demonstrated the efficacy of mibefradil in lowering blood pressure and as an antianginal and antiischemic agent. At clinically recommended doses, mibefradil has a heart rate lowering effect without a negative inotropic effect, and a favorable side effect profile. Because it is metabolized by the cytochrome P450 pathway, it should be used cautiously with other agents similarly metabolized. PMID- 9725541 TI - Teaching clinical pharmacology and therapeutics: selective for fourth-year medical students. AB - Teaching clinical pharmacology remains both a lifelong learning process and a lifelong challenge for clinical pharmacologists and other medical educators. In the current information age, with an explosion of drug-related data, the prime topic for discussion is how to teach clinical pharmacology. This article describes our response to the challenges in developing a selective course in clinical pharmacology, and our experience from the first 2 years of the course. Our emphasis is on how to provide in an efficient way knowledge, skills, and attitudes students will need as physicians in the coming decades. Faculty from the Center for Clinical Pharmacology at the University of Pittsburgh in conjunction with faculty from the University of Pittsburgh School of Medicine have developed a one-month intensive course in clinical pharmacology. The integrated course program consists of four overlapping components: 1) general clinical pharmacology (focused on individualization of drug therapy); 2) rational prescribing principles (general principles of drug selection, how to prepare a personal formulary); 3) disease-specific clinical topics (pharmacotherapy of diseases and medical conditions most commonly seen in routine medical practice); and 4) workshops for special attention topics (pharmacokinetics, pain treatment, toxicology, dialysis). In congruence with established educational goals, the course includes drug-, patient-, and disease-oriented concepts. A variety of learning formats (didactic and interactive lectures, one-day problem-based learning sessions, small group case discussions, self-directed and small group directed learning, quizzes, and computer-assisted learning) are used to teach students how to apply the general concepts of clinical pharmacology and rational pharmacotherapy to clinical medicine, and to prepare them to become independent lifelong learners in therapeutics. Student feedback from the first 2 years of this course indicates that this multi-modal teaching format is effective. The majority of students who took the course in clinical pharmacology in 1997 found it to be very beneficial. PMID- 9725542 TI - Assessment of drug accumulation in the evaluation of pharmacokinetic data. AB - The evaluation of drug accumulation is approached from a practical point of view. Estimates of accumulation indices as obtained from standard estimators-AUC, peak levels, and trough levels (RAUAUC Rmax and Rmin, respectively)-are compared and differences analyzed. The estimators are based on the concentration-time curve characteristics area under the concentration-time curve (AUC), maximum concentration, and trough level. Simulated data are used for the analysis, both noise-free and with random error added. The data are based on pharmacokinetic parameters derived from a clinical study. The numerical procedures can be reproduced by the interested reader with little effort. It is shown empirically that if Rmin, > RAUC then simple kinetic behavior cannot be safely assumed, but accumulation is a complex function of time. Rmax as obtained from the data and an estimate of this value based on time to peak concentration (tmax) and apparent elimination rate constant (lambda(z)) after a single dose and at steady state can then be compared in an attempt to exclude time-dependent kinetics. This new numerical procedure can provide valuable and even pivotal information regarding the accumulation kinetics of a compound under investigation. Recommendations on how to use the available concentration-time information to the best advantage are presented. It is concluded that the assessment of drug accumulation should not be confined to the calculation of just one estimate, because the three estimators RAUC, Rmax. and Rmin reflect different aspects of accumulation. Moreover, all information about accumulation should be carefully analyzed in the clinical context. This way the relevant accumulation can be identified for safe and efficacious drug treatment. PMID- 9725543 TI - Pharmacokinetics and pharmacodynamics of zolmitriptan in patients with mild to moderate hypertension: a double-blind, placebo-controlled study. AB - Zolmitriptan is a potent selective 5HT1B/1D receptor agonist for acute migraine therapy. Zolmitriptan has vasoconstrictor activity in cerebral vessels and may cause slight elevations of blood pressure in subjects without hypertension. Therefore, the pharmacokinetics and pharmacodynamics of zolmitriptan (5, 10, and 20 mg) were evaluated in 16 patients with mild to moderate hypertension (controlled by hydrochlorothiazide 50 mg once daily) and 17 healthy age- and sex matched control subjects in a randomized, placebo-controlled, double-blind, four period crossover study. The pharmacokinetics of zolmitriptan and its metabolites were dose proportional. Although area under the concentration-time curve (AUC0 infinity) and maximum concentration (Cmax) were slightly higher in patients with hypertension at all doses, this was only statistically significant for AUC at the 20-mg dose. Differences between subjects with and without hypertension were not clinically significant. Zolmitriptan produced a small increase in blood pressure, but this was similar in subjects with and without hypertension and was of no clinical significance. Zolmitriptan was well tolerated in both groups. Zolmitriptan plasma concentrations were higher in women than in men, with higher values of AUC and Cmax and lower total clearance in women. These results indicate that zolmitriptan can be administered for treatment of migraine in patients with controlled hypertension without dose adjustment. PMID- 9725544 TI - Effect of hepatic impairment on the pharmacokinetics of zolmitriptan. AB - Zolmitriptan, an oral 5HT1D agonist for the acute treatment of migraine, is cleared from the systemic circulation mainly by hepatic metabolism. Consequently, changes in hepatic function may result in changes in the pharmacokinetics of zolmitriptan. This open, parallel-group study was conducted to compare the pharmacokinetics and tolerability of a single 10-mg dose of zolmitriptan in healthy subjects and patients with hepatic impairment. A total of 37 participants entered and completed the study, including 10 healthy volunteers, 11 patients with moderate hepatic impairment, 10 patients with severe hepatic impairment without ascites, and 6 patients with severe hepatic impairment with ascites. The metabolism of zolmitriptan was reduced in patients with severe hepatic impairment compared with healthy subjects, resulting in higher peak plasma concentrations (47%), increased exposure (226%), and prolonged half-life (157%). The changes were similar in the presence and absence of ascites. Smaller changes were observed in patients with moderate hepatic impairment. Plasma concentrations of the three major metabolites of zolmitriptan were reduced in the patients with hepatic impairment. Patients with moderate hepatic impairment require no dosage adjustment, but the recommended daily intake of zolmitriptan may need to be reduced in patients with severe hepatic impairment. PMID- 9725545 TI - Oral bioavailability and disposition characteristics of irbesartan, an angiotensin antagonist, in healthy volunteers. AB - Absolute oral bioavailability and disposition characteristics of irbesartan, an angiotensin II receptor antagonist, were investigated in 18 healthy young male volunteers. Subjects received [14C] irbesartan as a 30-minute intravenous infusion (50 mg), [14C] irbesartan orally as a solution (50 mg or 150 mg), or irbesartan capsule (50 mg). Irbesartan was rapidly and almost completely absorbed after oral administration, and exhibited a mean absolute oral bioavailability of 60% to 80%. Mean total body clearance was approximately 157 mL/min, and renal clearance was 3.0 mL/min. Volume of distribution at steady state was 53 L to 93 L, and terminal elimination half-life was approximately 13 to 16 hours. Hepatic extraction ratio was low (0.2). There were no major circulating metabolites, and approximately 80% of total plasma radioactivity was attributable to unchanged irbesartan. Regardless of route of administration, approximately 20% of dose was recovered in urine and the remainder in feces. PMID- 9725546 TI - Pharmacokinetics of torsemide in patients with decompensated and compensated congestive heart failure. AB - Plasma pharmacokinetics of oral furosemide have been shown to be influenced by degree of decompensation in patients with congestive heart failure (CHF). This open-label, sequential comparison trial was conducted to determine whether CHF decompensation also alters the pharmacokinetics and pharmacodynamics of torsemide. Twelve patients with CHF, defined by either hemodynamic parameters or clinical signs and symptoms, were enrolled. On admission for treatment of their CHF, the patients were given 100 mg oral torsemide (phase A). A second dose of oral torsemide 100 mg was administered after hemodynamic parameters and clinical signs and symptoms of decompensated CHF resolved (phase B). Plasma and urine samples were collected over a 24-hour period for determination of torsemide concentrations and urine sodium. Hemodynamic measurements and physical signs and symptoms also were evaluated. During phase A, patients had significantly greater urine output and fractional sodium excretion compared with phase B. A significant increase in the area under the plasma concentration-time curve (AUC) was observed during phase B compared with phase A. However, no significant differences in maximal excretion rate of torsemide were noted between phase A and phase B. Heart failure status slightly affects the plasma pharmacokinetics of torsemide; however, this does not significantly alter the maximal urinary excretion rate of torsemide. PMID- 9725547 TI - Influence of age on the safety, tolerability, and pharmacokinetics of the novel HMG-CoA reductase inhibitor cerivastatin in healthy male volunteers. AB - The safety, tolerability, and pharmacokinetics of cerivastatin, a novel, synthetic, potent, and highly selective HMG-CoA reductase inhibitor, were studied in 48 young and elderly male volunteers in a randomized, double-blind, placebo controlled study. Eight men ranging from 18 to 38 years of age (young) and 15 men ranging from 65 to 78 years of age (elderly) received 0.1-mg cerivastatin tablets daily for 7 days. The remaining subjects (8 young and 17 elderly) received matching placebo tablets. Cerivastatin was well tolerated in elderly and young subjects. Adverse events were mild and occurred less frequently in the participants receiving cerivastatin than in those receiving placebo. In those participants given cerivastatin, the incidence of adverse events was similar for both age groups (4 of 8 young subjects and 8 of 15 elderly subjects). Transient and mild elevations in creatine kinase and transaminase levels were evenly distributed across the cerivastatin and placebo groups. Pharmacokinetic parameters, including area under the concentration curve (AUC), peak plasma concentration (Cmax), time to Cmax (tmax), and elimination half-life (t1/2), were similar between the two age groups. The mean elimination t1/2 for both groups was approximately 4 hours. These results indicate that cerivastatin is well tolerated in elderly male volunteers at a dosage of 0.1 mg/day. Further, the pharmacokinetics of cerivastatin are not altered as a consequence of age. Dose adjustment is therefore not required in elderly men. PMID- 9725548 TI - Pharmacokinetics and pharmacodynamics of scopolamine after subcutaneous administration. AB - The effects of subcutaneously administered scopolamine on quantitative electroencephalogram (qEEG) and cognitive performance were evaluated and correlated with pharmacokinetic parameters in a randomized, double-blind placebo controlled crossover study of 10 healthy male volunteers. Changes in qEEG and cognition were determined for 8 hours after drug administration. Scopolamine produced dose- and time-dependent impairments of attention and memory and a time dependent increase in delta power (1.25-4.50 Hz) and a decrease in fast alpha power (9.75-12.50 Hz) on qEEG compared with placebo. Maximum serum concentrations of scopolamine occurred 10 to 30 minutes after drug administration. Mean peak serum concentrations (free base) were 3.27, 8.99, and 18.81 ng/mL after administration of 0.4, 0.6 mg, and 0.8 mg scopolamine, respectively. Elimination half-life was approximately 220 minutes. The findings indicate temporary changes in qEEG and psychometric tests, and support the possible use of such a testing model for impaired cognitive functions such as age-related memory disturbances. PMID- 9725549 TI - Plasma protein binding of letrozole, a new nonsteroidal aromatase enzyme inhibitor. AB - The protein binding characteristics of the nonsteroidal aromatase inhibitor letrozole were determined using 14C-labeled letrozole. The binding of letrozole in human serum was 60.1 +/- 2.9% as a mean obtained in six individual sera and was similar in human plasma. The binding in human serum remained constant at concentrations of letrozole ranging from 10 to 500 ng/mL. A similar binding value in human serum was obtained using equilibrium dialysis and ultrafiltration technique. Albumin (binding 55.1 +/- 1.4%) is the main protein involved in the drug binding to plasma proteins. Increases in letrozole concentration (10-500 ng/mL) had no effect on binding. Albumin binding appeared to be nonsaturable with a binding capacity of 2 L/mmol. Binding to alpha1-acid glycoprotein and to gamma globulins was lower than 10%. The fraction in erythrocytes with a hematocrit of 0.4 was found to be 35.2 +/- 2.7%. Letrozole binding to serum proteins of rat, dog, mouse, and rabbit was approximately 10% lower than that in human serum and approximately 20% lower than that in baboons. Tamoxifen (100-1,500 ng/mL) had no effect on the in vitro binding of letrozole. Ex vivo binding in plasma from patients after repeated administration of letrozole alone (61.4 +/- 2.6%) was the same as after combined administration of letrozole and tamoxifen (60.0 +/- 3.2%). PMID- 9725550 TI - Safety, pharmacokinetics, and antiretroviral activity of the potent, specific human immunodeficiency virus protease inhibitor nelfinavir: results of a phase I/II trial and extended follow-up in patients infected with human immunodeficiency virus. AB - The safety, antiretroviral activity, and pharmacokinetic profile of nelfinavir, a potent and specific inhibitor of human immunodeficiency virus (HIV) protease, were assessed in a small open-label phase I/II dose-ranging study in protease inhibitor-naive HIV-positive men. A total of 22 patients with baseline plasma HIV RNA > or = 20,000 copies/mL and CD4+ counts between 200 and 500 cells/mm3 were enrolled in the study. Of the 22 patients, 20 were evaluated for activity; 10 patients assigned to 771 mg/day base equivalent (300 mg three times daily) and 10 patients assigned to 1,026 mg/day base equivalent (600 mg twice daily) given monotherapy. A capsule formulation of nelfinavir was used. The initial study period was 28 days; patients showing a virologic response of 1 log10 reduction were eligible for enrollment in an extension phase and addition of nucleoside analogues. A maximally tolerated dose of nelfinavir was not established. A dose response relationship was observed for four (40%) patients in the 771-mg group and six (60%) patients in the 1,026-mg group experiencing a reduction from baseline in plasma HIV RNA of at lest 1 log during the 28-day study. Of these patients, five sustained the reduction in plasma HIV RNA beyond day 28 (2 patients receiving 771 mg/day and 3 patients receiving 1,026 mg/day). Median increases from baseline in CD4+ counts at day 28 were 216 cell/mm3 and 86 cell/mm3 in the 771-mg and 1,026-mg groups, respectively. After oral administration, median nelfinavir plasma concentrations on day 28 reached a maximum at 1 hour (2,966 ng/mL) in the 771-mg group and at 3 hours (3,157 ng/mL) in the 1,026-mg group. Data for 22 patients were included in the safety analysis; 12 patients (55%) reported at least one grade 2 or worse (moderate, severe, or very severe) adverse event. The most common grade 2 or worse adverse event was diarrhea, reported by two patients (20%) receiving 771 mg/day and seven patients (70%) receiving 1,026 mg/day; followed by nausea, flatulence, asthenia, and headache (each reported in 1 patient [10%] in the 771-mg group) and dizziness (reported in 1 patient [10%] receiving 1,026 mg/day). In the small subgroup (n = 6) who continued taking nelfinavir for longer periods (between 8 and 15 months), virologic responses were sustained in the majority of patients with good tolerability. Nelfinavir is an active HIV-protease inhibitor with favorable pharmacokinetics, good tolerability, and sustained antiviral effects. Results of this early phase I/II dose-ranging study provided data for the safety and antiretroviral activity of nelfinavir and led to the selection of higher doses for phase II/III trials to further optimize virologic and immunologic responses. PMID- 9725551 TI - Metabolic disposition of 14C-bromfenac in healthy male volunteers. AB - The metabolic disposition of 14C-bromfenac, an orally active, potent, nonsteroidal, nonnarcotic, analgesic agent was investigated in six healthy male subjects after a single oral 50-mg dose. The absorption of radioactivity was rapid, producing a mean maximum plasma concentration (Cmax) of 4.9 +/- 1.8 microg x equiv/mL, which was reached 1.0 +/- 0.5 hours after administration. Unchanged drug was the major component found in plasma, and no major metabolites were detected in the plasma. Total radioactivity recovered over a 4-day period from four of the six subjects averaged 82.5% and 13.2% of the dose in the urine and feces, respectively. Excretion into urine was rapid; most of the radioactivity was excreted during the first 8 hours. Five radioactive chromatographic peaks, a cyclic amide and four polar metabolites, were detected in 0- to 24-hour urine samples. Similarity of metabolite profiles between humans and cynomolgus monkeys permitted use of this animal model to generate samples after a high dose for structure elucidation. Liquid chromatography/mass spectrometry (LC/MS) analysis of monkey urine samples indicated that the four polar metabolites were two pairs of diastereoisomeric glucuronides whose molecular weight differed by two daltons. Enzyme hydrolysis, cochromatography, and LC/MS experiments resulted in the identification of a hydroxylated cyclic amide as one of the aglycones, which formed a pair of diastereoisomeric glucuronides after conjugation. Data also suggested that a dihydroxycyclic amide formed by the reduction of the ketone group that joins the phenyl rings formed the second pair of diastereoisomeric glucuronides. Further, incubation of various reference standards in control (blank) urine and buffer with and without creatinine indicated that the hydroxy cyclic amide released from enzyme hydrolysis can undergo ex vivo transformations to a condensation product between creatinine and an alpha-keto acid derivative of the hydroxy cyclic amide that is formed by oxidation and ring opening. Further experiments with a dihydroxylated cyclic amide after reduction of the keto function indicated that it too can form a creatinine conjugate. PMID- 9725552 TI - Atorvastatin does not produce a clinically significant effect on the pharmacokinetics of terfenadine. AB - The effect of atorvastatin, a CYP3A4 substrate, on the pharmacokinetics of terfenadine and its carboxylic acid metabolite, fexofenadine, were evaluated. Single 120-mg doses of terfenadine were given 2 weeks apart to healthy volunteers with 80-mg daily doses of atorvastatin administered from 7 days before through 2 days after the second terfenadine dose. Concentrations of terfenadine and fexofenadine were measured for 72 hours after each terfenadine dose. Administration of terfenadine alone or in combination with atorvastatin produced no alterations in the QTc interval. For terfenadine, atorvastatin coadministration produced an 8% decrease in maximum concentration (Cmax), a 35% increase in area under the concentration-time curve extrapolated to infinity (AUC0-infinity), and a 2% decrease in elimination half-life (t1/2). For fexofenadine, atorvastatin coadministration produced a 16% decrease in Cmax, a 2% decrease in AUC0-infinity and a 51 % increase in t1/2. None of these changes achieved statistical significance. Coadministration of atorvastatin with terfenadine does not result in a clinically significant drug interaction. Because 80 mg is the highest atorvastatin dose used clinically, drug interactions mediated by CYP3A4 inhibition are unlikely in clinical practice. PMID- 9725554 TI - Advances in the automated detection of metaphase chromosomes labeled with fluorescence dyes. AB - Applications of fluorescence in situ hybridization (FISH) for translocation studies and biological dosimetry would benefit substantially from reliable and efficient automatic detection of metaphase chromosomes labeled with fluorescent dyes. We replicated and evaluated a fluorescence metaphase finder previously developed at the Medical Research Council (MRC), Human Genetics Unit (Scotland) and at Lawrence Berkeley Laboratory (LBL; California). The MRC/LBL system seemed to detect nearly all of the metaphases on the test slides, but it presented an unacceptable number of false positives (about five false positives per one true positive). Furthermore, we determined that the system actually overcalled true detections by counting certain metaphase spreads twice (duplicates). Through modifications of the MRC/LBL system, we developed the Lawrence Livermore National Laboratory (LLNL) system, which minimizes the detection of duplicates, incorporates new detection features, uses a binary decision tree (BDT) for classification, and provides functionalities to improve scanning accuracy and improve the post-detection review. To test the new system, DAPI-stained preparations of metaphase chromosomes from blood lymphocytes of four unrelated donors were placed on slides in drops ranging from 7 mm to 20 mm in diameter. Drops contained between 5 and 200 scorable metaphases each. The LLNL system achieved approximately 90% detection of non-duplicated metaphases as verified by an expert cytogeneticist, with typically less than one false positive per every one true positive detected. PMID- 9725553 TI - Enzyme kinetic reactions and fluorochrome uptake rates measured in individual cells by laser scanning cytometry. AB - This study was designed to explore the utility of a microscope-based cytofluorometer, the laser scanning cytometer (LSC), for time-resolved kinetic measurements. This instrument measures fluorescence of individual cells rapidly, with high sensitivity and accuracy. Also recorded in a list mode fashion are the spatial X-Y coordinates of the cell on the slide as well as the time of individual cell measurement. Repeated measurement of each of a group of cells within a selected area of the slide, thus, yields information on their fluorescence parameters (integrated value, maximal pixel intensity, or fluorescence area) as a function of time. Using the fluorogenic substrate di (leucyl)-rhodamine 110, we measured the kinetic activity of L-aminopeptidase in HL-60 cells and in monocytes, granulocytes, and lymphocytes from human blood. Likewise, the rate of fluorescein diacetate (FDA) hydrolysis by esterases was measured in HL-60 cells. Also studied was the rate of uptake of the lysosomo trophic fluorochrome acridine orange (AO) by human leukocytes. Several hundred cells per sample were measured rapidly with a time resolution of 20-60 s. The resolution was inversely proportional to the number of cells within the measured population. The kinetic curves constructed for individual cells had clearly defined slope and plateau regions. During data analysis the kinetic plots were matched with the respective cells; the latter were identified by their position on the slide and classified by their fluorescence or image after staining with Giemsa. Great intercellular variability was noted in enzyme kinetics among cells of the same type, and differences were seen in rate of AO uptake between granulocytes, monocytes, and lymphocytes. Fluorescence fading and recovery was observed especially in the case of FDA hydrolysis and AO uptake. Our results indicate that LSC can be used to rapidly measure the rate of uptake or accumulation of a particular fluorochrome or the kinetics of enzymatic reactions in individual cells of large populations to reveal intercellular variability or the presence of a cell subpopulation with different kinetic properties. PMID- 9725555 TI - Single-cell measurement of superoxide anion and hydrogen peroxide production by human neutrophils with digital imaging fluorescence microscopy. AB - Besides flow cytometry, fluorescence microscopy combined with computerized image analysis offers an alternative tool for assessing phagocyte oxidant generation at the single-cell level. This technique provides an opportunity for the direct visualization of cells and simultaneous measurement of cellular fluorescence intensity. Thus, we developed a simple method for the quantitative evaluation of intracellular superoxide anion and hydrogen peroxide production with image cytometry by using hydroethidine and dihydrorhodamine 123 dyes, respectively. Human neutrophils stimulated with phorbol dibutyrate and labeled by these fluorogenic substrates showed intense, well recognizable red or green fluorescence. The intensity of signals from individual granulocytes of cytospin preparations were quantitatively measured in digitized images. There was a great heterogeneity in response to the stimulus within the granulocyte population as shown by the integrated fluorescence intensity values. In agreement with the results of parallel flow cytometric experiments, this simple image analysis performed on cells of cytospin preparations was able to detect the defects in the oxidative metabolism of neutrophils from patients with cervix carcinoma. We demonstrated that even minor alterations in superoxide anion/hydrogen peroxide generation can be detected by image cytometry as efficiently as by flow cytometry. This result validates imaging microscopy as an alternative to flow cytometry in such experiments. In addition, the image cytometric technique allows the observation of the kinetics of free radical production in individual cell under adherent conditions. Therefore, we carried out image analysis of the oxidative burst of neutrophils adherent to uncoated glass and fibronectin- and type IV collagen-coated surfaces in response to stimulation with phorbol dibutyrate or N-formyl-methionyl-leucyl-phenylalanine. We elaborated a calibration technique for the quantitative measurement of the ethidium bromide generation mediated by superoxide anion within individual adherent granulocytes. The ethidium bromide production varied between 0.48 and 1.17 amol/cell/min. PMID- 9725556 TI - Automated breast tumor diagnosis and grading based on wavelet chromatin texture description. AB - In this paper, wavelets were employed for multi-scale image analysis to extract parameters for the description of chromatin texture in the cytological diagnosis and grading of invasive breast cancer. Their value was estimated by comparing the performance of co-occurrence, densitometric, and morphometric parameters in an automated K-nearest neighbor (Knn) classification scheme based on light microscopic images of isolated nuclei of paraffin-embedded tissue. This design allowed a multifaceted cytological retrospective study of which the practical value can be judged easily. Results show that wavelets perform excellently with classification scores comparable with densitometric and co-occurrence features. Moreover, because wavelets showed a high additive value with the other textural groups, this panel allowed a very profound description with higher recognition scores than previously reported (76% for individual nuclei, 100% for cases). Morphometric parameters performed less well and only slightly increased correct classification. The major drawback, besides image segmentation errors demanding operator supervision, emanated to be the few false-negative cases, which restrict the immediate practical use. However, an enlargement of the parameter set may avoid this misclassification, resulting in an applicable expert system of practical use. PMID- 9725557 TI - Fluorescence-imaging assay for cytotoxic plaque formation and for growth toward confluency of adherent cells. AB - A nondestructive fluorescence-imaging assay is described for quantitating the number and size of plaques formed over time by cytotoxic effector cells in a monolayer of target cells. It can also be used to assay the growth of adherent cells toward confluence. The method involves the use of fluorescein conjugated to high molecular weight dextran. The dextran is excluded by adherent cells, thereby making the medium around cells more fluorescent than the cells themselves. The area of the plate that is fluorescent can be determined by confocal fluorescence imaging microscopy. With this new method, changes in the confluency of adherent cells or in the number and area of cytotoxic plaques can be assayed repeatedly over an extended period of time, without manipulation of the cells or of the medium. PMID- 9725558 TI - Cell cycle effects and induction of apoptosis caused by infection of HL-60 cells with human granulocytic ehrlichiosis pathogen measured by flow and laser scanning cytometry. AB - Human granulocytic ehrlichiosis (HGE) is an occasionally severe and even fatal disease caused by an agent closely related to Ehrlichia equi and Ehrlichia phagocytophila, which is transmitted by ticks. Little is known about the pathogen itself, which only very recently has been isolated. The agent can be cultivated in vitro because it replicates in human promyelocytic leukemic HL-60 cells. Using multiparameter flow cytometry and laser scanning cytometry (LSC) we have investigated changes in HL-60 cells following their infection with the pathogen. Its presence within the infected HL-60 cells was detected and its intracellular level measured inmmunocytochemically using antibodies obtained from HGE-infected patients. The percentage of the infected cells measured by flow cytometry or LSC correlated well with the estimates by microscopy on the Giemsa-stained specimens. In the infected cultures, the cells had diminished levels of cyclins D3 and E as well as the cyclin dependent kinase inhibitor p21WAF1/CIP1 and were arrested predominantly in G0/1. The apoptosis-associated regulatory proteins were also affected by cell infection: expression of Bcl-2 was decreased in the infected cells whereas expression of Bax become more variable, with some cells showing higher levels of this protein. The infected cells developed numerous DNA strand breaks characteristic of apoptosis. The presence of the pathogen was also detected by LSC in cells from peripheral blood of the infected patients; after relocation and visual inspection ("CompuSort") the pathogen-positive cells were identified as leukocytes. This unique ability of LSC to detect, quantify, and visualize HGE in infected cells made this instrument particularly useful to measure the degree of infection in peripheral blood of the patients and study effects of the infectious agent on the cell cycle and apoptosis of the host cells. PMID- 9725559 TI - Cell cycle effects of CB30865, a lipophilic quinazoline-based analogue of the antifolate thymidylate synthase inhibitor ICI 198583 with an undefined mechanism of action. AB - CB30865 (p-[N-(7-bromo-3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl+ ++)-N (prop-2-ynyl)amino]-N-(3-pyridylmethyl)benzamide) is a quinazoline-based pyridine containing compound that emerged from a programme aimed at the development of thymidylate synthase (TS) inhibitors as anticancer agents. Its structure is based on the antifolate ICI 198583, but with a pyridine ring replacing the glutamate. Despite its structure, CB30865 does not act in vitro via inhibition of TS or, apparently, other known folate-dependent pathways, and extensive mechanistic studies suggest that it acts via a novel locus with respect to conventional antitumour agents. However, CB30865 is highly potent against a variety of human tumour cell lines (e.g., 50%-inhibitory concentration [IC50] values in the 1-10 nM range). Thus, the cell cycle effects of CB30865 were investigated. DNA histogram analysis of W1L2 human lymphoblastoid, L1210 murine leukaemia, and CH1 human ovarian cells (propidium iodide staining) has demonstrated that CB30865 does not cause a phase-specific arrest at concentrations that have been shown to inhibit colony formation. This is unexpected for an anticancer agent. In W1L2 cells, using bromodeoxyuridine (BrdUrd) labelling and bivariate Hoechst/ propidium iodide staining, it was revealed that 0.003-0.15 microM CB30865 (1-50 x 72 h IC50) caused cells to arrest in all phases of the cell cycle simultaneously after 20-24 h exposure. This effect was also observed in CH1 and L1210 cells, though the arrest was at slightly different times. Thus, using this technique, it has been demonstrated that CB30865 induces an unusual and delayed cell cycle arrest, which provides further evidence for a novel locus of action for this compound. PMID- 9725561 TI - Comparison of two flow cytometric methods enumerating CD4 T cells and CD8 T cells. AB - The enumeration of blood CD4+ (single-positive) T cells by flow cytometry is subject to errors such as counting CD4+ monocytes or CD4+CD8+ (double-positive) T cells as CD4+ T cells. Relatively accurate count can be obtained when CD4+ T cells are defined as CD3+CD4+CD8- mononuclear cells (MNCs), using 3-color flow cytometry. However, using this approach, further classification into CD4+ T cell subsets requires expensive 4-color flow cytometry. In an attempt to enumerate CD4+ T cells using only 2 colors, we have defined CD4+ T cells as MNCs expressing CD4 and not expressing the markers of other MNCs (CD8, CD13, CD14, and CD16) and compared the results to those obtained with the 3-color method in which CD4+ T cells were defined as CD3+CD4+CD8- MNCs. Both methods produced similar results. An analogous approach was undertaken to enumerate CD8+ T cells as MNCs expressing CD8 and not expressing the markers of other MNCs. However, when compared with the percents obtained by the 3-color method in which CD8+ T cells were defmed as CD3+CD4-CD8+ MNCs, the 2-color method overestimated the percent of CD8+ T cells. This was likely due to counting CD8low null cells as CD8+ T cells. When the percentages of CD8high T cells were evaluated, both methods produced similar results. We conclude that the 2-color method is suitable for the enumeration of CD4+ T cells and CD8high T cells and reserves the third color for further enumeration of CD4+ or CD8high T cell subsets. PMID- 9725560 TI - Flow cytometric characterization of proliferating natural killer lymphocytes from bone marrow donors in the mixed lymphocyte reaction. AB - Detection of natural killer (NK) cells in the mixed lymphocyte reaction (MLR) was investigated by flow cytometry and cytotoxicity toward the K562 cell line. Peripheral blood mononuclear cells (PBMCs) from normal individuals were stimulated with either lymphoblastoid cell lines (LCLs) or PBMCs. This approach allowed the following observations: 1) after stimulation by LCLs, the percentage of NK cells increased concomitantly with the level of cytotoxicity, in contrast to when PBMCs were used as stimulators; 2) anti-interleukin-2 monoclonal antibody strongly inhibited NK proliferation in the MLR, but antibody to interferon gamma did not; and 3) purified NK cells were unable to proliferate against LCLs. All these data suggest that NK cells induced by LCL stimulators depend on T cells to proliferate in the MLR. Flow cytometric detection of NK cells in the MLR was also used on cryopreserved PBMCs from 31 bone marrow donors, because it has been reported that an enhanced donor NK cell activity was correlated with the development of graft-versus-host disease after HLA-identical sibling bone marrow transplantation. NK cell proliferation under LCL stimulation was intense and varied greatly among the donors tested. However, no statistical correlation was observed between LCL-induced donor NK cell proliferation in the MLR and the occurrence of graft-versus-host disease after bone marrow grafting. PMID- 9725562 TI - Specificity of binding of HIV-1 anti-p24 antibodies to CD4+ lymphocytes from HIV infected subjects. AB - The clinical utility of a flow cytometric assay (FCA) for intracellular HIV p24 antigen was evaluated in a group of HIV-1-infected subjects. A previously described method, p24-FCA (1), was modified to minimize nonspecific staining and to include irrelevant isotype-matched control antibodies. Blood mononuclear cells from 32 HIV-1 seropositive subjects and 14 HIV-1 seronegative controls were examined. In HIV-1 seropositive individuals, the proportion of CD4+ lymphocytes that bound the p24 monoclonal and the isotype control antibodies were not different. The frequency of cells binding p24 antibodies increased with declining CD4 counts and was highest in patients with low CD4+ lymphocyte counts. Although results of p24-FCA do reflect disease progression, the high nonspecific binding of monoclonal antibodies in infected subjects obscures specific p24 binding and precludes its use as an accurate measure of infection within single cells. PMID- 9725563 TI - Rapid detection of rod photoreceptor apoptosis by flow cytometry. AB - Apoptosis is a form of cell death that plays an important role during physiological homeostasis and numerous pathological conditions. Retinitis pigmentosa is a group of retinal degenerative disorders in which rod photoreceptors excessively die via apoptosis. Current analytical tools for the investigation of photoreceptor cell death are histological in nature and typically time-consuming and laborious; as such, they are frequently limited to small sample sizes. In recent years, apoptosis research has made extensive use of flow cytometry to analyze several cellular events characteristic of apoptosis, including DNA strand nicking and cell shrinkage. This study presents a flow cytometric assay for the detection of rod photoreceptor apoptosis using the rod specific cell marker rhodopsin, the terminal deoxyuridine 5'-triphosphate (dUTP) transferase nick end-labeling (TUNEL) assay, and the light-scattering properties of rod photoreceptors. This technique enables the rapid and reproducible detection of rod photoreceptor apoptosis in vitro with potential applications for in vivo analysis. PMID- 9725564 TI - Immunohistochemical localization of fibroblast growth factors (FGFs) and FGF receptor-1 in human normal salivary glands and pleomorphic adenomas. AB - Basic and acidic fibroblast growth factors (bFGF and aFGF) are heparin-binding growth factors, and promote fibrogenesis and angiogenesis. We investigated the immunohistochemical localization of bFGFE, aFGF, and FGF receptor-1 in pleomorphic adenomas. In the normal salivary glands, bFGF was localized in the basement membranes of intercalated ducts, acini and basal cells of the excretory ducts, while aFGF was localized focally in the intercalated ductal cells and basal cells of the excretory ducts. In pleomorphic adenomas, bFGF was immunolocalized in the basement membranes around the solid nests of myoepithelial cells, around the neoplastic myoepithelial cells in the myxoid areas, and in the lacuna cells in the chondroid areas. In contrast, chondroid areas exhibited no immunoreactivity with aFGE. Positive signals for aFGF were localized in luminal cells of the tubuloglandular structures in pleomorphic adenomas. FGF receptor-1 immunolocalized in the lacuna cells and myoepithelial cells in the solid and myxoid areas. These observations suggest that bFGF and FGF receptor-1 produced by myoepithelial cells inhibited terminal differentiation and enchondral ossification in pleomorphic adenomas. These results also suggest important roles for FGFs in the formation of various structures with mesenchymal-like histology. PMID- 9725565 TI - Immunohistochemical demonstration of bone morphogenetic protein-2 and type II collagen in pleomorphic adenoma of salivary glands. AB - Immunohistochemical investigation of bone morphogenetic protein-2 (BMP-2) and type II collagen, two cartilage-associated proteins, was undertaken using monoclonal antibodies in 20 cases of salivary pleomorphic adenoma (PA) in order to explore their possible roles in chondroid differentiation of this tumor. Other salivary gland tumors, including adenoid cystic carcinoma (17 cases), polymorphous low-grade adenocarcinoma (10 cases), basal cell adenoma (3 cases), basal cell adenocarcinoma (1 case), and epithelial-myoepithelial carcinoma (2 cases), were also examined for comparison. In PA, BMP-2 immunoreactivity was detected in the luminal and non-luminal cells of the tubulo-ductal structures, plasmacytoid cells, and other scattered tumor cells in solid areas. In addition, tumor cells in chondroid areas in most cases (14/15), and stellate cells in myxoid areas in many cases (7/19), were also intensely labeled for BMP-2. Furthermore, BMP-2 was also detected in the non-neoplastic ductal cells in salivary glands, whereas no other salivary gland tumors were positively stained for this protein. Type II collagen was localized in the intercellular matrix of chondroid areas and in a few chondroid differentiating cells in myxoid areas, confirming its cartilage-specificity. A proportional relationship was observed between BMP-2 expression and chondroid formation, although BMP-2 was also stained in occasional PAs without chondroid formation. It is speculated that BMP-2 might be secreted by tumor cells and play a role in chondroid formation in PA by inducing some tumor cells to produce type II collagen and other chondroid matrical substances, like glycosaminoglycans. The expression of BMP-2 is specific to PA and may possibly be used as a useful marker in differentiating PA from other salivary gland tumors. PMID- 9725566 TI - Loss of heterozygosity at 5q21-22 (adenomatous polyposis coli gene region) in oral squamous cell carcinoma is common and correlated with advanced disease. AB - We determined the frequency of loss of heterozygosity (LOH) at chromosome 5q21-22 (adenomatous polyposis gene region) in oral SCC from 49 patients using PCR-based assays. Of 43 informative (heterozygous) tumors, 41.9% [95% confidence interval (CI)=27.0, 57.9] contained LOH at 5q21-22. LOH at 5q21-22 was strongly associated with stage at diagnosis: 100%, (3/3), 50% (13/26), and 14% (2/14) of tumors from patients with distant metastases, regional spread, and localized disease, respectively, contained this genetic alteration (P=0.01). There were no statistically significant associations between LOH at 5q21-22 and other patient or tumor characteristics, but LOH was more commonly found in the tumors of heavy smokers, infrequent alcohol consumers, and in tumors containing either p53 mutations or HPV-DNA. In univariate analyses, LOH at 5q21-22 was associated with poor prognosis (hazard ratio=1.8, 95%, CI 0.8, 4.5); this relationship did not persist after adjustment for stage of disease (hazard ratio=1.1, 95% CI=0.4, 3.1). These data provide further evidence that inactivation of the APC gene and/or other genes at 5q21-22 is common and may be involved in the development and/or progression of oral SCC. Larger studies are needed to determine whether LOH at 5q21-22 is linked to known oral SCC etiologic factors and/or the prognosis of oral SCC patients, as well as to genetic instability at other loci involved in these malignancies. PMID- 9725567 TI - Expression of CD44 variant exons by primary and metastatic oral squamous carcinomas. AB - Abnormal CD44 expression in many neoplasms correlates with behaviour, but reports on its role in oral squamous carcinoma are contradictory. CD44 expression was characterised in a closely matched series of oral carcinomas with and without metastases in both frozen and formalin-fixed tissue and correlated with behaviour and histological grading parameters. Eleven primary oral squamous carcinomas without metastases and nine primary carcinomas with 19 matched metastases were stained immunocytochemically for CD44H and products of variant exons v3, v4/5, v6 and v9. Patterns of staining in frozen and formalin-fixed tissue were correlated with invasive front grading and behaviour using exact inferential statistics. Most primary carcinomas stained for all exons tested but some showed loss of expression of v4/5. Loss of expression was more marked in metastases, but there was no correlation between expression and behaviour or grade. Stromal surfaces of epithelial cells often expressed variant exon products reflecting loss of polarity. This, together with selective loss of v4 and v5 in primary carcinomas and their more frequent loss in metastases, suggests that CD44 may play a role in metastasis of some oral squamous carcinomas. PMID- 9725568 TI - Expression of cadherins and catenins in oral epithelial dysplasia and squamous cell carcinoma. AB - The immunocytochemical expression of cadherins and catenins was examined during the process of oral carcinogenesis by comparing their expression in normal and dysplastic epithelium with primary and metastatic carcinomas. While control epithelium showed normal distribution for P and E cadherin and the catenins, in severe dysplasia P-cadherin was upregulated. In other cases and in carcinoma-in situ adjacent to infiltrating carcinomas, membranous expression of the cadherins and catenins was reduced or lost. The changes in expression of E-cadherin and the catenins suggest that disruption of the E-cadherin/catenin complex is a late event associated with invasion. In primary carcinomas reduced membranous and cytoplasmic staining were observed for both cadherins and catenins. Abnormal localisation of E-cadherin occurred in the more superficial parts of the better differentiated carcinomas, suggesting abnormality to the E-cadherin complex(es). In contrast, membranous expression of cadherins and catenins was reduced or lost in the deep invasive margin of primary carcinomas and in most poorly differentiated carcinomas. For E-cadherin at least, this reduction appears associated with differentiation, invasion and possibly prognosis. Possible mechanisms involved for changes in expression of the cadherins and associated catenins and areas for further study are discussed. PMID- 9725569 TI - Epithelial p53 gene expression and mutational analysis, combined with growth fraction assessment, in oral lichen planus. AB - The immunohistochemical detection of epithelial p53 protein expression in oral lichen planus (OLP) biopsies was supplemented with molecular analysis for mutations of the p53 gene using the polymerase chain reaction - single stranded conformational polymorphism (PCR-SSCP) technique. p53 protein expression, in the basal epithelial cell layer, as detected by the DO7 and 1801 antibodies, was significantly more frequent in OLP compared with other oral keratoses and normal mucosa, as was the growth fraction. The 10 OLP biopsies that had the most frequent p53 staining (plus a case of OLP found in continuity with a SCC) were screened by the PCR-SSCP technique, but no mutations were detected in the p53 gene (exons 5 9). The p53 overexpression in the OLP samples may be a physiological response to the hyper-proliferative state, as revealed by the growth fraction determination. This may usefully serve to protect against mutagenesis, and so be a factor in the low incidence of carcinoma associated with OLP. PMID- 9725571 TI - Multiple calcifying hyperplastic dental follicles. AB - Multiple calcifying hyperplastic dental follicles is a rare condition characterized by multiple unerupted teeth with abundant calcifications and rests of odontogenic epithelium in their enlarged dental follicles. This article describes an additional example of this condition. PMID- 9725570 TI - Adhesion of oral C. albicans to human buccal epithelial cells following limited exposure to antifungal agents. AB - The major aetiologic agent of oral candidosis is C. albicans, and adhesion to oral mucosal surfaces is considered a vital prerequisite for successful colonisation and subsequent infection by this agent. Although many antimycotics are available for the treatment of oral candidosis, the diluent effect of saliva and the cleansing action of the oral musculature often tend to reduce the availability of the agents below that of the effective therapeutic concentration. Therefore, the yeasts undergo only a limited exposure to the antifungals during therapy. Hence, the main aim of the present study was to determine the in vitro adhesion of ten isolates of oral C. albicans to buccal epithelial cells (BEC) following a short exposure to sublethal concentrations of four antifungal agents. The yeasts were exposed to sublethal concentrations of nystatin (x6 MIC), 5 fluorocytosine (x8 MIC), ketoconazole (x4 MIC) and fluconazole (x4 MIC) for a period of 1 h. Following subsequent removal of the drug, the adhesion of these isolates to BEC was assessed by a previously described adhesion assay. The mean percentage reductions of candidal adhesion to BEC following exposure to sublethal concentrations of nystatin, 5-fluorocytosine, ketoconazole and fluconazole were 72.88%, 16.52%, 40.16% and 24.36%, respectively. Ultrastructural studies revealed that short exposure to nystatin and the azoles (but not 5-fluorocytosine) resulted in aberrant cellular features. These findings indicate that subtherapeutic levels of antifungals may modulate candidal colonisation of the oral mucosa and thereby suppress the invasive potential of the pathogen. PMID- 9725572 TI - Variations of the uncinate process of the lateral nasal wall with clinical implications. AB - The morphology of the uncinate process (UP) and nasal fontanelle is described in 119 human specimens, which were examined both before and after removal of the mucosa. Forms of the UP are classified and based on which site the process is articulated, and each form is characterized in relation to the endonasal endoscopic operative technique. Type I: The infero-posterior tip of the UP is articulated to the inferior concha (turbinate). This was the most frequently observed type. Subtype I-b: The UP adhered to the inferior concha along the antero-inferior margin. The anterior nasal fontanelle was closed by the UP adhesion; therefore, special attention is required not to damage the lacrimal bone. Type N: The tip of the UP had no articulation and made a free edge. It reduces the bony resistance at surgery. Type S: The tip articulated to the superior structures, such as the bulla ethmoid, medial orbital wall, tegument of the maxillary sinus, and basal area of the ethmoid sinus. These structures are known as high-risk areas of endonasal surgery (Levine, 1993). Type P: The tip articulated with the perpendicular plate of the palatine bone. The UP was prolonged posteriorly. Attention should be paid to the sphenopalatine artery, which goes through the posterior edge of the middle concha. Four additional variations (combinations of the above basic types, Variations IS, IP, SP, and ISP) were also observed. PMID- 9725573 TI - The topographic relationships of the unpaired visceral branches of the aorta. AB - We examined the metric relationship among the origins of the unpaired visceral branches of the aorta, their relationship to the total descending aorta (TDA), and the relationship between the TDA and stature to see whether a graft for the TDA, e.g., from the left subclavian artery to the aortic bifurcation, which includes its visceral branches, could be pre-constructed. These proportions were compared between the genders and between adults and children to see whether any differences exist. Thirty-four adult aortae and eight juvenile aortae were examined. These segments-TDA, aortic bifurcation to celiac artery, aortic bifurcation to superior mesenteric artery, and aortic bifurcation to inferior mesenteric artery, were defined as the distances between the origins of the left subclavian, celiac, superior mesenteric, and inferior mesenteric arteries, respectively, to the aortic bifurcation. Stature was known only in 15 adult cadavers. The absolute lengths of the segments were correlated to each other and the ratios of these absolute lengths (proportional lengths) were calculated. The statistical significance was examined by Student's T-test and variability by the F test. The TDA correlated well with aortic bifurcation to celiac artery and aortic bifurcation to superior mesenteric artery, whereas a weaker correlation existed with aortic bifurcation to inferior mesenteric artery. The ratio aortic bifurcation to celiac artery and aortic bifurcation to superior mesenteric artery to TDA was less variable than the ratio aortic bifurcation to inferior mesenteric artery to TDA. The abdominal aorta measured approximately a one-third of TDA. No differences in correlation nor in ratio were found between genders and between adults and children. No correlation between stature and TDA was found. The two upper unpaired visceral branches originate from the aorta in a prefixed site, which correlates closely with the length of the descending aorta, whereas the lower one has a more variable point of origin. This is true for all ages and for both genders. Aortic length does not correlate with stature. It is not possible thus to predict the length of the descending aorta by stature. However, a model of the descending aorta can be constructed, but with less accuracy, for the inferior mesenteric artery. PMID- 9725574 TI - Double or bifid zygomaticus major muscle: anatomy, incidence, and clinical correlation. AB - The anatomy of the double or bifid zygomaticus major muscle is investigated in a series of 50 hemifacial cadaver dissections. The double zygomaticus major muscle represents an anatomical variation of this muscle of facial expression. This bifid muscle originates as a single structure from the zygomatic bone. As it travels anteriorly, it then divides at the sub-zygomatic hollow into superior and inferior muscle bundles. The superior bundle inserts at the usual position above the comer of the mouth. The inferior bundle inserts into the modiolus below the corner of the mouth. The incidence of the double zygomaticus major muscle was 34% in the present study, as it was found to be present in 17 of 50 cadaver dissections. This study shows that variation in the individual morphology of the mimetic muscles can be a common finding. Clinically, the double or bifid zygomaticus major muscle may explain the formation of cheek "dimples." The inferior bundle was observed in several specimens to have a dermal attachment along its mid-portion, which tethers the overlying skin. When an individual with this anatomy smiles, traction on the skin may create a dimple due to this dermal tethering effect. PMID- 9725575 TI - Quantification of antagonist muscle coactivation in children with spastic diplegia. AB - The purpose of this study was to investigate the use of the Pearson Product Moment correlation coefficient to quantify muscle coactivation using electromyography. The subjects were two children with spastic diplegia. Surface electrodes were used to record muscle activity from the soleus and tibialis anterior muscles during voluntary attempts at dorsiflexion and plantarflexion of the ankle against resistance. The linear envelope signals were smoothed with a Hamming filter at a cutoff frequency of 4 Hz and intervals of the resultant pairs of curves subjected to the PPM test. Both subjects demonstrated significant coactivation on their right side and independent activity on the left, indicated by high positive and negative coefficients, respectively. This method shows promise for description of side differences in diplegics and for assessing the effects of physical therapy and other interventions. PMID- 9725576 TI - Mechanical strain at the pisotriquetral joint. AB - Pain and tenderness on the palmar and ulnar aspects of the wrist in the area of the pisiform bone is fairly common. Chronic pain in the pisiform area may be due to tendinitis of the insertion of the flexor carpi ulnaris, bony fractures or osteoarthrosis of pisotriquetral joint which is the second most common degenerative arthritis in wrist after the scaphotrapezial osteoarthrosis (Fischer, 1988, Radiologe 28:338-344). Subperiostal excision of pisiform bone is customarily performed after unsuccessful initial non-operative treatment (Carroll and Coyle, 1985, J. Hand Surg. 10:703-707; Johnston and Tonkin, 1986, Clin. Orthop. 210:137-142; Nuesch et al., 1993, Handchir. Mikrochir. Plast. Chir. 25:42 45). Although the postoperative results seem to be rather good, possible malfunction based on excision has not been considered previously by investigators. The aim of this study was to improve our understanding of the role of the pisiform and the pisotriquetral joint in the transfer of forces within the carpus. In a first part we examined 112 pisotriquetral joints by qualitative, quantitative and densitometrical analysis of joint surfaces. Secondly, mechanical tests were performed to investigate the distribution of forces within pisiform and the pisotriquetral joint. The results demonstrate that the pisiform mechanically contributes to the stability of the ulnar column of the wrist. The pisiform, and its unique anatomical holding mechanism, discharges two main functions. It holds the triquetrum in a correct position and prevents its subluxation even in extreme extension. Furthermore, it acts as a fulcrum (hypomochlion) while transducing powerful forearm forces to the hand. The excision of pisiform should be reconsidered. PMID- 9725577 TI - Classification of communications between the musculocutaneous and median nerves. AB - In 16 out of 79 cadavers 22 communications were found between the musculocutaneous and median nerves. In six subjects they were present bilaterally. There were three types, based on the sites of communication. Type I: The communication was proximal to the entrance of the musculocutaneous nerve into coracobrachialis (9/22); Type II: The communication was distal to the muscle (10/22); Type III: The nerve as well as the communicating branch did not pierce the muscle (3/22). Bilateral communications were not necessarily of the same type. The possible clinical implications of these communications (relating either to the surgical approach to the shoulder joint, or to entrapment syndromes) are discussed. PMID- 9725578 TI - The pterygoideus propius muscle revisited. AB - The pterygoideus proprius muscle is an anomaly in the infratemporal fossa that has immovable attachments to the infratemporal crest and the lateral pterygoid plate of the sphenoid bone. We report the identification of three cases of this anomaly, suggest its embryological origin, and speculate about its function. PMID- 9725579 TI - Magnetic resonance tomographic angiography of the arterial circle (of Willis). PMID- 9725580 TI - Problem in diagnostic imaging: a patient with sciatica. AB - This article presents the imaging findings in a patient with sciatica. The reader is invited to identify the labeled anatomical structures and the lesions present. The discussion focuses on the anatomy of the lumbar spine and the relative merits of the various methods of imaging a patient with sciatica. PMID- 9725581 TI - Anomalous thymic branch of the right common carotid artery. AB - A previously unreported anomalous thymic artery that branched from the anterior aspect of right common carotid artery approximately 1 cm above bifurcation of the brachiocephalic artery was found during routine dissection. It traveled inferiorly through a plexus of inferior thyroid veins for 6 cm in front of the brachiocephalic artery and crossed the anterior surface of the trachea where it divided into two branches that supplied the right and left lobes of the thymus. The development and blood supply of the thymus and their clinical anatomy are reviewed. PMID- 9725582 TI - The nature of the superior sinus venosus defect. AB - The location, and morphology, of the superior sinus venosus interatrial communication remains contentious. As part of a clinical study, we examined anatomic specimens and echocardiograms so as to clarify the arrangement of the normal atrial septal structures, and compared them with the arrangement found in the superior sinus venosus defect. The pathognomonic diagnostic criterion in the abnormal hearts was overriding of the intact muscular rim of the oval fossa by the mouth of the superior caval vein. This muscular rim is, in reality, a tube of myocardium which encloses a core of extracardiac adipose tissue. Understanding of this anatomic conundrum clarifies the understanding of the structures of both the normal atrial septum and sinus venosus defects. PMID- 9725583 TI - Is inguinal hernia a defect in human evolution and would this insight improve methods of surgical repair. PMID- 9725584 TI - The forest and the trees. PMID- 9725585 TI - An experimental evaluation of recent electron dosimetry codes of practice. AB - As from the 1 January 1997, the recent IPEMB code of practice for electron dosimetry is the recommended protocol for electron beam dosimetry in the UK, replacing the previous HPA code of practice and its IPSM addendum. New recommendations for electron beam dosimetry have also been formulated recently by the AAPM and the IAEA on the use of parallel-plate ionization chambers in high energy electron beams. Against this background, the procedures recommended in each of these codes of practice have been followed from intercomparison of the field instrument ionization chamber with a secondary standard through to the determination of absorbed dose at the reference position in the electron beam. Absorbed doses have been determined for a number of electron beam energies ranging from nominal 5 MeV through to 17 MeV, and for four different types of field instrument ionization chamber: an NE2571 graphite walled cylindrical chamber; an NACP parallel-plate chamber; a Markus parallel-plate chamber; and a Roos parallel-plate chamber. The differences in the determination of absorbed dose between the IPEMB protocol and the HPA/IPSM protocol vary from +0.5% to +1.6% at the depth of maximum dose. In addition the IPEMB measured doses are 0.2% larger than those measured following the IAEA code of practice. It may also be stated that the IPEMB measured doses at the depth of maximum dose are up to 1.5%, but generally less than 1.0%, lower than those measured by the AAPM protocol. PMID- 9725586 TI - A Monte Carlo study of the quality dependence factors of common TLD materials in photon and electron beams. AB - A Monte Carlo simulation of the quality dependence of different TL materials, in the form of discs 3.61 mm in diameter and 0.9 mm thick, in radiotherapy photon beams relative to 60Co gamma-rays has been performed. The beam qualities ranged from 50 kV to 25 MV x-rays. The TL materials were: CaF2, CaSO4, LiF and Li2B4O7. The effects of the dopants on energy deposition in the TL material have also been determined for the highly sensitive LiF:Mg:Cu:P (TLD-100H) and for CaF2:Mn. It was found that there was a significant difference in the quality dependence factor derived from Monte Carlo simulations between LiF and LiF:Mg:Cu:P but not between CaF2 and CaF2:Mn. The quality dependence factors for Li2B4O7 varied from 0.990 +/- 0.008 (1 sd) for 25 MV x-rays to 0.940 +/- 0.009 (1 sd) for 50 kV x rays relative to 60Co gamma-rays; Monte Carlo simulations were also performed for Li2B4O7 in megavoltage electron beams. For CaF2, the quality dependence factor varied from 0.927 +/- 0.008 (1 sd) for 25 MV x-rays to 10.561 +/- 0.008 (1 sd) for 50 kV x-rays. The figure for CaSO4 ranged from 0.943 +/- 0.008 (1 sd) for 25 MV x-rays to 9.010 +/- 0.008 (1 sd) for 50 kV x-rays. The quality dependence factor for CaF2 increases by up to 5% with depth and by up to 15% with field size for the kilovoltage x-ray beams. For LiF-TLD, however, there was no significant dependence on the field size or depth of irradiation in the kilovoltage energy range. PMID- 9725587 TI - Ion recombination corrections for the NACP parallel-plate chamber in a pulsed electron beam. AB - The NACP electron chamber is one of three parallel-plate chambers recommended for use in the UK. Measurements with this chamber type have indicated a problem in determining the recombination correction. This is due to a variation of the ionization current I with polarizing voltage V which deviates from the accepted Boag theory. It is shown that there is a chamber-dependent threshold voltage below which the NACP chamber follows the Boag theory. Above this voltage the chamber should be used with caution, although it is still possible to correct for the dependence of the chamber response on the dose per pulse. The existence of such deviations from theory demonstrates the usefulness of the 1/I against 1/V plot and the limitations of the Boag two-voltage analysis. Values for the initial recombination and the coefficient of general recombination are measured for several NACP chambers. It is shown that from these one can derive a value for the effective plate separation and the collector radius of each chamber. Differences in the behaviour of NACP chambers manufactured by Scanditronix and Dosetek are discussed and the implications of free-electron collection are considered. PMID- 9725588 TI - Accurate portal dose measurement with a fluoroscopic electronic portal imaging device (EPID) for open and wedged beams and dynamic multileaf collimation. AB - Measuring portal dose with an electronic portal imaging device (EPID) in external beam radiotherapy can be used to perform routine dosimetric quality control checks on linear accelerators and to verify treatments (in vivo dosimetry). An accurate method to measure portal dose images (PDIs) with a commercially available fluoroscopic EPID has been developed. The method accounts for (i) the optical 'cross talk' within the EPID structure, (ii) the spatially nonuniform EPID response and (iii) the nonlinearity of the EPID response. The method is based on a deconvolution algorithm. Measurement of the required input data is straightforward. The observed nonlinearity of the EPID response was largely due to the somewhat outdated EPID electronics. Nonlinearity corrections for more modern systems are expected to be smaller. The accuracy of the method was assessed by comparing PDIs measured with the EPID with PDIs measured with a scanning ionization chamber in a miniphantom, located at the same position as the fluorescent screen. For irradiations in open, wedged and intensity modulated 25 MV photon beams (produced with dynamic multileaf collimation) EPID and ionization chamber measurements agreed to within 1% (1 SD). PMID- 9725589 TI - Volume recombination parameter in ionization chambers. AB - The parameter m2 governing the volume recombination in ionization chambers has been measured under conditions which allow strict application of the basic theory. The method consists of measuring the ratio of ionization currents I(V1) and I(V2) obtained at two given voltages V1 and V2 as a function of I(V1). The value of m2 is derived from a linear extrapolation to zero current. Several pairs of voltages (V1, V2) were used. The value of m2 obtained in this work is 3.97 x 10(14) s m(-1) C(-1) V(2) with a relative uncertainty of 1.7%. The dependence of m2 on atmospheric conditions has also been investigated. PMID- 9725590 TI - Thermoluminescent response and relative efficiency of TLD-100 exposed to low energy x-rays. AB - The dose-response of LiF:Mg,Ti (TLD-100) exposed to 15 and 35 kVp (8.0 +/- 0.1 and 8.1 +/- 0.1 keV effective energy respectively) x-rays and 60Co gamma-rays has been measured in the dose interval from (1.2-5.4) x 10(3) Gy for x-rays, and from 0.14 to 850 Gy for gamma-rays. In both cases the total TL signal and glow curve peaks 3 to 9 show supralinearity. The supralinearity function f(D) is similar for both x-ray beams, except for peak 8, where a 30% difference is observed. The maxima of f(D) for the total TL signal and peaks 5 to 8 are 2.1, 1.7, 6.4, 3.3 and 7.5 respectively for 8.1 keV x-rays and 3.7, 3.1, 13.6, 9.9 and 11.0 for gamma-rays. The measured relative efficiencies for x-rays with respect to 60Co, for the total TL signal and peaks 5 and 7, were 1.04, 0.97 and 3.2 respectively. PMID- 9725591 TI - Application of a diamond detector to brachytherapy dosimetry. AB - The feasibility of using a diamond detector for the dosimetry of brachytherapy sources has been investigated. A high-activity 192Ir source was selected for this purpose. The dosimetric characteristics measured included the photon fluence anisotropy in air, transverse dose profiles in planes parallel to the plane containing the HDR source and isodose distributions. The 'in-air' anisotropy of the photon fluence relative to seed orientation was measured at 5 and 10 cm from the source centre and compared with TLD measurements. Transverse dose distributions in planes parallel to the plane containing the source long axis were measured in a water phantom and compared with calculations performed with a treatment planning system. Isodose distributions were also measured in several planes around the 192Ir source. Measurements on two sources indicate that the 'in air' photon fluence anisotropy measured by the diamond detector and the TLDs is very similar. Dose profiles measured at several distances from the source are also found to be in good agreement with the calculated dose profiles and isodose distributions. Results of this feasibility study indicate that the diamond detector, with its excellent spatial resolution and nearly tissue equivalent and isotropic radiation response, is an appropriate detector for dose measurements around brachytherapy sources. PMID- 9725592 TI - In-water calibration of PDR 192Ir brachytherapy sources with an NE2571 ionization chamber. AB - An ionometric calibration procedure for 192Ir PDR brachytherapy sources in terms of dose rate to water is presented. The calibration of the source is performed directly in a water phantom at short distances (1.0, 2.5 and 5.0 cm) using an NE2571 Farmer type ion chamber. To convert the measured air-kerma rate in water to dose rate to water a conversion factor (CF) was calculated by adapting the medium-energy x-ray dosimetry protocol for a point source geometry (diverging beam). The obtained CF was verified using two different methods. Firstly, the CF was calculated by Monte Carlo simulations, where the source-ionization chamber geometry was modelled accurately. In a second method, a combination of Monte Carlo simulations and measurements of the air-kerma rate in water (at 1.0, 2.5 and 5.0 cm distance) and in air (1 m distance) was used to determine the CF. The obtained CFs were also compared with conversion factors calculated with the adapted dosimetry protocol for high-energy photons introduced by Tolli. All calculations were done for a Gammamed PDR 192Ir source-NE2571 chamber geometry. The conversion factors obtained with the four different methods agree to within 1% at the three distances of interest. We obtained the following values (medium energy x-ray protocol): CF(1 cm) = 1.458; CF(2.5 cm) = 1.162; CF(5.0 cm) = 1.112 (1 sigma = 0.7% for the three distances of interest). The obtained results were checked with TLD measurements. The values of the specific dose rate constant and the radial dose function calculated in this work are in accordance with the literature data. PMID- 9725593 TI - Developing a dose-volume histogram computation program for brachytherapy. AB - A dose-volume histogram (DVH) computation program was developed for brachytherapy treatment planning in an attempt to benefit from the DVH's ability to present graphically information on 3D dose distributions. The program is incorporated into a planning system that utilizes a pair of orthogonal radiographs to localize the radiation sources. DVHs are calculated for the volume of tissue enclosed by an isodose surface (e.g. half the value of the reference isodose). The calculation algorithm is based on a non-uniform random sampling that gives a denser point distribution at the centre of the implants. Our program was tested and proved to be fast enough for clinical use and sufficiently accurate (i.e. computation time of 20 s and less than 2% relative error for one point source, for 100,000 calculation points). The accuracy improves when a larger calculation point number is used, but the computation time also increases proportionally. The DVH is presented in the form of a simple graph or table, or as Anderson's 'natural' DVH graph. The cumulative DVH tables can be used to extract a series of indexes characterizing the homogeneity and the dose levels of the distribution in the treatment volume and the surrounding tissues. If a reference plan is available, the DVH results can be assessed relative to the reference plan's DVH. PMID- 9725594 TI - A case study comparing the relative benefit of optimizing beam weights, wedge angles, beam orientations and tomotherapy in stereotactic radiotherapy of the brain. AB - A treatment-planning case study has been performed on a patient with a medium sized, convex brain tumour. The study involved the application of advanced treatment-plan optimization techniques to improve on the dose distribution of the 'standard plan' used to treat the patient. The standard plan was created according to conventional protocol at the Royal Marsden NHS Trust, and consisted of a three-field (one open and two wedged) non-coplanar arrangement, with field shaping to the beam's-eye view of the planning target volume (PTV). Three optimized treatment plans were created corresponding to (i) the optimization of the beam weights and wedge angles of the standard plan, (ii) the optimization of the beam orientations, beam weights and wedge angles of the standard plan, and (iii) a full fluence tomotherapy optimization of 1 cm wide (at isocentre), 270 degree arcs. (i) and (ii) were created on the VOXELPLAN research 3D treatment planning system, using in-house developed optimization algorithms, and (iii) was created on the PEACOCK tomotherapy planning system. The downhill-simplex optimization algorithm is used, in conjunction with 'threshold-dose' cost function terms enabling the algorithm to optimize specific regions of the dose volume histogram (DVH) curve. The 'beam-cost plot' tool is presented as a visual aid to the selection of beneficial beam directions. The methods and pitfalls in the transfer of plans and patient data between the two planning systems are discussed. Each optimization approach was evaluated, relative to the standard plan, on the basis of DVH and dose statistics in the PTV and organs at risk (OARs). All three optimization approaches were able to improve on the dose distribution of the standard plan. The magnitude of the improvement was greater for the optimized beam-orientation and tomotherapy plans (up to 15% and 30% for the maximum and mean OAR doses). A smaller improvement was observed in the beam weight and wedge-angle optimized plan (up to 5% and 10% in the maximum and mean OAR doses). In the tomotherapy plan, difficulty was encountered achieving an acceptable homogeneity of dose in the PTV. This was improved by treating the gross tumour volume (GTV) and (PTV - GTV) regions as separate targets in the inverse planning, with the latter region prescribed a slightly higher dose to reduce edge under-dosing. In conclusion, for the medium-sized convex tumour studied, the tomotherapy dose distribution showed a significant improvement on the standard plan, but no significant improvement over a conventional three-field plan where the beam orientations, beam weights and wedge angles had been optimized. PMID- 9725595 TI - Calculation of the spatial variation of relative biological effectiveness in a therapeutic proton field for eye treatment. AB - The relative biological effectiveness (RBE) of protons under conditions suitable for eye treatment has been studied. A complete three-dimensional modelling of the beam delivery system has been performed. Proton Monte Carlo transport calculations have been performed to obtain the proton energy distributions at different positions in a water phantom including the influence of range shifter, modulator wheel, scattering foils and collimators. A beam with a kinetic energy of 68 MeV +/- 250 keV has been simulated with respect to the HMI-Berlin eye treatment facility. The dependence of the RBE on absorbed dose and position within a spread-out Bragg peak (SOBP) has been investigated with the track structure model. Due to a decreasing proton energy with depth, the energy transfer per pathlength within the SOBP increases, affecting the RBE. An RBE increasing with depth as well as with decreasing absorbed dose has been found for the endpoint inactivation of V79 and CH2B2 hamster cells. RBE values at the distal end of the SOBP up to 1.3 and 1.5 have been found at a dose of 14 Gy and 2 Gy respectively. Within the SOBP plateau, no lateral variation of RBE has been found for a given depth. The model used offers the possibility of introducing a variable RBE in treatment planning. PMID- 9725596 TI - Predicting the radiation control probability of heterogeneous tumour ensembles: data analysis and parameter estimation using a closed-form expression. AB - A closed-form formula describing the tumour control probability (tcp) of a heterogeneous collection of tumours has been obtained by analytically averaging the homogeneous double-exponential tcp formula over inter-tumour distributions of clonogen radiosensitivity, density and repopulation rate, tumour volume and dose. The formula can be straightforwardly and relatively quickly fitted to clinical data, yielding radiobiological parameter values for use in tcp modelling. The formula was fitted to published tcp data which catalogued tumour control records grouped by dose and tumour volume, and treatment duration. Fitted parameter values, confidence intervals and goodness-of-fit statistics were determined. The sets of parameter values obtained are unique only to within a scaling factor. The formula provides non-rejectable fits to data which grouped tcp by dose and volume when radiosensitivity parameters take values close to laboratory estimates, the fitted volume dependence parameter, however, taking rather high values. Good fits are obtainable with the intuitively reasonable volume parameter value of one, but with radiosensitivity values around one-third of their laboratory estimates. Non rejectable fits to data which grouped tcp by dose and treatment duration may be obtained with radiosensitivity and repopulation rate parameters lying close to laboratory estimates. PMID- 9725597 TI - Optimization of beam orientation in radiotherapy using planar geometry. AB - This paper proposes a new geometrical formulation of the coplanar beam orientation problem combined with a hybrid multiobjective genetic algorithm. The approach is demonstrated by optimizing the beam orientation in two dimensions, with the objectives being formulated using planar geometry. The traditional formulation of the objectives associated with the organs at risk has been modified to account for the use of complex dose delivery techniques such as beam intensity modulation. The new algorithm attempts to replicate the approach of a treatment planner whilst reducing the amount of computation required. Hybrid genetic search operators have been developed to improve the performance of the genetic algorithm by exploiting problem-specific features. The multiobjective genetic algorithm is formulated around the concept of Pareto optimality which enables the algorithm to search in parallel for different objectives. When the approach is applied without constraining the number of beams, the solution produces an indication of the minimum number of beams required. It is also possible to obtain non-dominated solutions for various numbers of beams, thereby giving the clinicians a choice in terms of the number of beams as well as in the orientation of these beams. PMID- 9725598 TI - Quality assurance of central axis dose data for photon beams by means of a functional representation of the tissue phantom ratio. AB - A recently proposed four-parameter functional representation for the tissue phantom ratio (TPR) in the domain of electronic equilibrium was tested for accuracy and applied to quality assurance of central axis dose data. The four parameters are energy dependent and are found for each beam by a minmax search. For photon energies of 4 MV and greater, the functional representation was accurate to within 1% of measured data for all depths and field sizes exceeding 10 cm. The representation was also found to be robust. With only nine measurements of the TPR (at the extremes and middle of the electronic equilibrium domain) used to determine the four parameters, the representation reproduced the TPR value of any depth and field size to within 1% of measured data for photon energies of 6 MV or greater. The representation was insensitive to random measurement errors of a magnitude comparable to that expected in clinical practice. Other findings indicated that the representation degrades gracefully as the domain is extended into regions not in electronic equilibrium. When used with a QA program, the functional representation of TPR provides a means of detecting and correcting errors of measurement and data transcription of central axis dose data. PMID- 9725599 TI - Quantitative evaluation of 2 x 2 arrays of Lucite cone applicators in flat layered phantoms using Gaussian-beam-predicted and thermographically measured SAR distributions. AB - SAR distributions from four different E-field-orientated 2 x 2 arrays of incoherently driven Lucite cone applicators (LCAs) were investigated. The LCAs operated at 433 MHz with an aperture of 10.5 cm x 10.5 cm each. Two techniques were used to obtain SAR distributions in flat layered phantoms: Gaussian beam (GB) predictions and thermographical (TG) imaging. The GB predictions showed that the effective field size of the different array configurations varied by up to 3%. The TG-measured SAR distribution showed significant deviations from the GB predicted SAR distributions (maximum 34.6%). The difference between GB-predicted and TG-measured SAR levels (averaged per 10% GB-predicted SAR intervals) equalled less than 11.3% for the parallel E-field orientated array and respectively 15.1% for the clockwise-orientated array. When antennae in the clockwise-orientated array were more widely spread (array aperture 23 cm x 23 cm) in order to diminish their mutual interactions, these differences decreased to 12.4%. However, the overall difference within the 50% SAR or higher range decreased from 14% to 9%. The results lead us to conclude that LCAs can be used clinically and their antenna interactions are not considered to be a problem under clinical conditions. PMID- 9725600 TI - A thin target approach for portal imaging in medical accelerators. AB - A new thin-target method (patent pending) is described for portal imaging with low-energy (tens of keV) photons from a medical linear accelerator operating in a special mode. Low-energy photons are usually produced in the accelerator target, but are absorbed by the target and flattening filter, both made of medium- or high-Z materials such as Cu or W. Since the main contributor to absorption of the low-energy photons is self-absorption by the thick target through the photoelectric effect, it is proposed to lower the thickness of the portal imaging target to the minimum required to get the maximum low-energy photon fluence on the exit side of the target, and to lower the atomic number of the target so that predominantly photoelectric absorption is reduced. To determine the minimum thickness of the target, EGS4 Monte Carlo calculations were performed. As a result of these calculations, it was concluded that the maximum photon fluence for a 4 MeV electron beam is obtained with a 1.5 mm Cu target. This value is approximately five times less than the thickness of the Cu target routinely used for bremsstrahlung production in radiotherapeutic practice. Two sets of experiments were performed: the first with a 1.5 mm Cu target and the second with a 5 mm Al target (Cu mass equivalent) installed in the linear accelerator. Portal films were taken with a Rando anthropomorphic phantom. To emphasize the low energy response of the new thin target we used a Kodak Min-R mammographic film and cassette combination, with a strong low-energy response. Because of its high sensitivity, only 1 cGy is required. The new portal images show a remarkable improvement in sharpness and contrast in anatomical detail compared with existing ones. It is also shown that further lowering of the target's atomic number (for example to C or Be) produces no significant improvement. PMID- 9725601 TI - Calculation of backscatter factors for diagnostic radiology using Monte Carlo methods. AB - Backscatter factors were determined for x-ray beams relevant to diagnostic radiology using Monte Carlo methods. The phantom size considered most suitable for calibration of dosimeters is a cuboid of 30 x 30 cm2 front surface and 15 cm depth. This phantom size also provides a good approximation to adult patients. Three different media were studied: water, PMMA and ICRU tissue; the source geometry was a point source with varying field size and source-to-phantom distance. The variations of the backscatter factor with phantom medium and field geometry were examined. From the obtained data, a set of backscatter factors was selected and proposed for adoption as a standard set for the calibration of dosimeters to be used to measure diagnostic reference doses. PMID- 9725602 TI - A new method of readout in radiochromic film dosimetry. AB - Radiochromic film as a dosimetry medium offers several advantages in high resolution radiography. A new technique of readout was developed to measure the optical density distributions of the film in purely directed light. This technique implements radiochromic film dosimetry near the film's absorption maximum by using a single-mode top-surface emitting laser diode (675.2 nm). The effective sensitivity of the film, compared with a helium-neon laser densitometer (632.8 nm), is increased approximately threefold. Good accuracy, high spatial resolution and simple assembly of the readout system is achieved. Beam profiles of the four final collimator helmets of a Leksell Gamma Knife (Elekta Inc., Sweden) were experimentally determined. Measured profiles and full-widths at half maximum are consistent with the computer generated data of the dose planning system (Kula 4.4, Elekta Inc., Sweden). The output factor of the 4 mm collimator (the smallest collimator with the steepest dose gradient), essential for the application of well defined doses, was checked. The measurements established an output factor of 826 +/- 9 that lies 9 +/- 1% lower than the adjusted one. PMID- 9725603 TI - Computer-aided radiation therapy simulation: image intensifier spatial distortion correction for large field of view digital fluoroscopy. AB - An accurate method of correcting spatial distortion in digital fluoroscopy images has been developed for generating fluoroscopy-based large field of view images for computer-aided radiation therapy simulation. This method is applicable to arbitrary gantry rotations and arbitrary shifts of the image intensifier relative to the central axis of the x-ray beam. It is therefore suitable for conventional radiation therapy simulation techniques that involve the arbitrary positioning of the image intensifier by the operator. Spatial distortion is modelled as two image intensifier orientation-dependent components, the first resulting from the projection of the x-ray image onto the curved surface of the image intensifier front end, and the second produced by the image intensifier electron optics, interactions with external magnetic fields and the video system. A geometrical model approximates the first component. The second component is modelled by a third-order polynomial transformation. A weighted mean approach is employed to achieve accurate distortion correction when the image intensifier is oriented differently from the calibration orientations. Mean and maximum residual errors (measured in the plane of the isocentre) of 0.4 mm and 1.0 mm respectively have been achieved with just 48 calibration orientations in four dimensions (gantry rotation and lateral, longitudinal and vertical shifts of the image intensifier). PMID- 9725604 TI - Clinical evaluation of a high-resolution new peripheral quantitative computerized tomography (pQCT) scanner for the bone densitometry at the lower limbs. AB - Precision, long-term stability, linearity and accuracy of the x-ray peripheral quantitative computerized tomographic (pQCT) bone scanner XCT 3000 (Norland Stratec Medical Sys.) were evaluated using the European Forearm Phantom (EFP). In vivo measurements were assessed using a standardized procedure at the distal femur and the distal tibia. In the patient-scan mode, the spatial resolution of the system was 1.04 +/- 0.05 lp/mm as measured at the 10% level of the modulation transfer function (MTF). The contrast-detail diagram (CDD) yielded a minimal difference in attenuation coefficient (AC) of 0.07 cm(-1) at an object size of 0.5 mm. The effective dose for humans was calculated to be less than 1.5 microSv per scan. Short-term precision in vivo was expressed as root mean square standard deviation of paired measurements of 20 healthy volunteers (RMSSD = 0.5%). At the distal femur total volumetric density (ToD) and total cross-sectional area (ToA) were found to be less sensitive to positioning errors than at the distal tibia. Structural parameters like the polar cross-sectional moment of inertia (CSMIp) or the polar cross-sectional moment of resistance (CSMRp) showed a good short-term precision at the distal femur (RMSSD = 1.2 and 1.4%). The relation between the two skeletal sites with respect to CSMIp or CSMRp showed a high coefficient of determination (r2 = 0.77 and 0.74). PMID- 9725605 TI - An investigation of pulse-timing techniques for broadband ultrasonic velocity determination in cancellous bone: a simulation study. AB - Berlage wavelets are used to simulate ultrasonic pulses in an unbounded, homogeneous, isotropic and absorptive medium. Intrinsic absorption of the medium is properly described by Kolsky's attenuation, which considers velocity dispersion to meet the causality condition. Several current time-domain velocity measurement techniques have been investigated using numerically simulated pulses for three normalized BUA values: 20, 40 and 60 dB MHz(-1) cm(-1), which mimic experimentally determined values for cancellous bone. The velocities, calculated using first motion transit times, are used as references supported by the Fermat principle of least time. The simulated results for fixed sample thickness indicate that pulse-broadening increases with the transit time of the reference point and the intrinsic absorption of the medium. Comparison shows that the first zero-crossing method yields 3-6% errors in velocity results, better than the cross-correlation method. However, the zero-crossing method gives inconsistent velocity measurement for a medium of 40 dB MHz(-1) cm(-1)1 absorption and three different thicknesses: 0.2, 0.4 and 0.6 cm. A novel technique for velocity measurement is presented using the peak of the envelope of a signal as a reference point to measure transit time difference. The envelope of a signal represents the instantaneous amplitude of the associated analytic signal. The velocities derived using this method differ from the true velocities by only 1.2 2.4%, more accurate than those obtained by the first zero-crossing method. The envelope peak has the additional merits of easy detection and robustness. Most importantly, the envelope technique may be used to yield accurate velocity measurement in cases where an accurate determination of the first motion transit time is sometimes prohibited due to the presence of noise. PMID- 9725606 TI - A rule based method for context sensitive threshold segmentation in SPECT using simulation. AB - Robust techniques for automatic or semi-automatic segmentation of objects in single photon emission computed tomography (SPECT) are still the subject of development. This paper describes a threshold based method which uses empirical rules derived from analysis of computer simulated images of a large number of objects. The use of simulation allowed the factors affecting the threshold which correctly segmented objects to be investigated systematically. Rules could then be derived from these data to define the threshold in any particular context. The technique operated iteratively and calculated local context sensitive thresholds along radial profiles from the centre of gravity of the object. It was evaluated in a further series of simulated objects and in human studies, and compared to the use of a global fixed threshold. The method was capable of improving accuracy of segmentation and volume assessment compared to the global threshold technique. The improvements were greater for small volumes, shapes with large surface area to volume ratio, variable surrounding activity and non-uniform distributions. The method was applied successfully to simulated objects and human studies and is considered to be a significant advance on global fixed threshold techniques. PMID- 9725607 TI - Evaluation of attenuation corrections using Monte Carlo simulated lung SPECT. AB - SPECT (single photon emission computed tomography) images are distorted by photon attenuation. The effect is complex in the thoracic region due to different tissue densities. This study compares the effect on the image homogeneity of two different methods of attenuation correction in lung SPECT; one pre-processing and one post-processing method. This study also investigates the impact of attenuation correction parameters such as lung contour, body contour, density of the lung tissue and effective attenuation coefficient. The Monte Carlo technique was used to simulate SPECT studies of a digital thorax phantom containing a homogeneous activity distribution in the lung. Homogeneity in reconstructed images was calculated as the coefficient of variation (CV). The isolated effect of the attenuation correction was assessed by normalizing pixel values from the attenuation corrected lung by pixel values from the lung with no attenuation effects. Results show that the CV decreased from 12.8% with no attenuation correction to 4.4% using the post-processing method and true densities in the thoracic region. The impact of variations in the definition of the body contour was found to be marginal while the corresponding effect of variations in the lung contour was substantial. PMID- 9725608 TI - Plane polarized x-ray fluorescence system for the in vivo measurement of platinum in head and neck tumours. AB - A plane polarized x-ray fluorescence system based on an orthovoltage radiotherapy treatment unit, the Pantak DXT-300, has been developed and optimized to measure tumour platinum concentration. The platinum derives from platinum based chemotherapy agents, such as cisplatin and carboplatin used to treat tumours in the head and neck region. Photons from an x-ray tube are polarized by scattering through 90 degrees, and used to stimulate the emission of characteristic platinum x-rays from the tumour. Information about the drug in the tumour could be of use in establishing dose-response relationships, in order to optimize the treatment and reduce the toxicity of the drug. The performance of the system was optimized with respect to the applied x-ray tube voltage, the current and the filter material; the effect on the minimum detection limit (MDL) of the thickness of the overlying tissue surrounding the tumour was investigated in detail. The lowest MDL is achieved using 0.25 mm of tin filter and x-ray tube voltage of 220 kV. This is 5.6 ppm for a tumour surrounded with 20 mm of overlying tissue, a measurement time of 2000 s and an estimated skin dose of 3.0 mGy. This represents the most sensitive in vivo XRF system to date. We have embarked on a clinical pilot study to measure the platinum concentration in tumours of the head and neck, and expect initial results to be available in the near future. PMID- 9725609 TI - Multiple ionization in the earlier stages of water radiolysis. AB - We have studied the fragmentation of water vapour molecules induced by collision with a Xe44+ beam at 6.7 MeV/u. From the measurement of the fragment time of flight, we show that the amount of fragmentation due to multiple ionization is very large. In the case of single ionization, we are able to reproduce accurately the experimental cross sections by calculating for each molecular level the single-ionization cross section in the framework of the CDW-EIS theory and with a diagram of dissociation modified with respect to the diagram obtained in the case of dipolar ionization. By using qualitative arguments based on the ability of the medium to neutralize a charged species, we tentatively extend our result to liquid water. From our analysis, we show that ionizations involving three or more ejected electrons could enhance the oxygen production. For the physicochemical phase we estimate that the rate of oxygen production by multiple ionization represents approximately 18% of the OH rate produced by single ionization. PMID- 9725610 TI - Power deposition in the head and neck of an anatomically based human body model for plane wave exposures. AB - At certain frequencies, when the human head becomes a resonant structure, the power absorbed by the head and neck, when the body is exposed to a vertically polarized plane wave propagating from front to back, becomes significantly larger than would ordinarily be expected from its shadow cross section. This has possible implications in the study of the biological effects of electromagnetic fields. Additionally the frequencies at which these resonances occur are not readily predicted by simple approximations of the head in isolation. In order to determine these resonant conditions an anatomically based model of the whole human body has been used, with the finite-difference time-domain (FDTD) algorithm to accurately determine field propagation, specific absorption rate (SAR) distributions and power absorption in both the whole body and the head region (head and neck). This paper shows that resonant frequencies can be determined using two methods. The first is by use of the accurate anatomically based model (with heterogeneous tissue properties) and secondly using a model built from parallelepiped sections (for the torso and legs), an ellipsoid for the head and a cylinder for the neck. This approximation to the human body is built from homogeneous tissue the equivalent of two-thirds the conductivity and dielectric constant of that of muscle. An IBM SP-2 supercomputer together with a parallel FDTD code has been used to accommodate the large problem size. We find resonant frequencies for the head and neck at 207 MHz and 193 MHz for the isolated and grounded conditions, with absorption cross sections that are respectively 3.27 and 2.62 times the shadow cross section. PMID- 9725611 TI - Neuromagnetic recordings of the human peripheral nerve with planar SQUID gradiometers. AB - Magnetic fields produced by a travelling volley in the human ulnar nerve have been successfully measured in a lightly shielded environment. Recordings of the tangential component of the magnetic field were made using a planar second-order gradiometer integrated with a first-order gradiometric superconducting quantum interference device (SQUID). Devices were fabricated in our clean-room facility at the University of Strathclyde and measurements taken in an eddy-current shielded room at the Wellcome Biomagnetism Unit. We use no additional shielding and no electronic differencing or field-nulling techniques. Evoked magnetic fields of 60 fT peak-to-peak were obtained after 1536 averages but they could be seen easily as early as 512 averages. Measurements were made over four points above the ulnar nerve on the upper arm and from these the conduction velocity was calculated as 60 m s(-1). PMID- 9725612 TI - A simple test device for electrometers. AB - Electrometers used in dosimetric instruments need to be checked regularly in order to measure and maintain the prescribed dose to the patient. This paper describes the function of a simple but accurate test device for electrometers. A number of electrometers have been tested and compared with calibrations performed by the Swedish Standard Dosimetry Laboratory (SSDL). The device can be used to test current, resistance, voltage and charge measurements. The charge can be conducted to the electrometer in different ways, and the input resistance of the electrometer can also be determined. The calibration factors obtained by the device are in good agreement with results obtained from calibrations at the SSDL. PMID- 9725613 TI - Gap-stepped MLC leaves with filler blades can eliminate tongue-and-groove underdoses when delivering IMRT with maximum efficiency. PMID- 9725614 TI - Thermal conductivity of uterine tissue in vitro. AB - Thermotherapy of the uterus has emerged as an alternative to hysterectomy in the treatment of menorrhagia, from whence it follows that the thermal properties of uterine tissue have become of importance. This study presents measurements of the thermal conductivity and the water content of uterine tissue in vitro. A steady state thermal conductivity apparatus, based on the comparison of test samples with a material with known thermal conductivity, is described. Measurements were conducted on tissue samples from eleven patients, directly after hysterectomy. Samples with and without endometrium, as well as coagulated samples, were examined. The thermal conductivity of myometrial tissue was found to be 0.536 +/- 0.012 W m(-1) K(-1) (mean +/- 1 SD) and the corresponding water content was 81.2 +/- 1.5% (mean +/- 1 SD). Measurements on samples with both endometrium and myometrium showed similar thermal conductivity (0.542 +/- 0.008 W m(-1) K(-1), mean +/- 1 SD) and water content (81.6 +/- 0.7%, mean +/- 1 SD). It was also indicated that coagulation causes dehydration, resulting in a lower thermal conductivity. PMID- 9725615 TI - Effects of read-out light sources and ambient light on radiochromic film. AB - Both read-out light sources and ambient light sources can produce a marked effect on coloration of radiochromic film. Fluorescent, helium neon laser, light emitting diode (LED) and incandescent read-out light sources produce an equivalent dose coloration of 660 cGy h(-1), 4.3 cGy h(-1), 1.7 cGy h(-1) and 2.6 cGy h(-1) respectively. Direct sunlight, fluorescent light and incandescent ambient light produce an equivalent dose coloration of 30 cGy h(-1), 18 cGy h(-1) and 0 cGy h(-1) respectively. Continuously on, fluorescent light sources should not be used for film optical density evaluation and minimal exposure to any light source will increase the accuracy of results. PMID- 9725616 TI - Combining various projection access schemes with the algebraic reconstruction technique for low-contrast detection in computed tomography. AB - We separately applied three different projection access orders (the multilevel scheme (MLS), the sequence access scheme (SAS) and the random permutation scheme (RPS)) to the algebraic reconstruction technique (ART) in an attempt to improve low-contrast object detection in computed tomography (CT). We used both simulated (from various numbers of projections of low-contrast and contrast detail phantoms) and actual data (from megavoltage portal CT) during our study. When coupled with ART, SAS and RPS led to poor low-contrast detection and required multiple iterations for convergence. In contrast, one-iteration MLS yielded the most uniform reconstructions with the highest contrast-to-noise ratios. Therefore, MLS-ART provides the highest dose efficiency of all current reconstruction techniques for imaging low-contrast objects from a small number of projections. PMID- 9725617 TI - Evaluation of the accuracy and precision of lung aerosol deposition measurements from planar radionuclide imaging using simulation. AB - Planar images of known, theoretical distributions of radioaerosol in the lung have been simulated using lung models derived from magnetic resonance studies on human subjects. Total lung activity was evaluated from the simulated images together with the absolute penetration index (PI) and a relative value expressed as a fraction of that in a simulated ventilation image. The accuracy and precision of these measurements were calculated by comparison with the true values used in the simulation. Total activity was assessed with systematic errors within 5% and precision within 6.5%. Measured PIs varied only slowly with true PI and inter-model variation masked changes between measurements on the different distributions. The relative PI reduced inter-model variation and provided significant differences between all the distributions. PI was significantly affected by misalignment of the lung region of interest. The conducting airways deposition fraction (CADF) used in the simulation correlated linearly with the fractional activity in a central lung region, allowing CADF to be estimated with a precision of 21%. PMID- 9725618 TI - Chameleon sequences in the PDB. AB - The Brookhaven Protein Data Bank has been searched for sequences that can be found both in helix and sheet conformation. The longest such sequences consist of seven residues. PMID- 9725620 TI - Prediction of an anti-IgE binding site on IgE. AB - A combination of computation techniques have been used to determine the binding site and amino acid residues on IgE that are critical for binding to a therapeutic anti-IgE. Homology modeling was used to model parts of IgE and of an antibody that binds to IgE. Docking simulations using shape descriptions were then carried out using the models to determine which residues in the IgE are involved in the binding interaction. The anti-IgE has been determined to bind close to some of the residues that are believed to be in the Fc(epsilon)RI receptor site on IgE, therefore preventing IgE from binding to Fc(epsilon)RI on mast cells and basophils and causing the release of pharmacologic mediators from these cells. Experiments have been suggested to verify the binding site and the residues involved in binding to anti-IgE. PMID- 9725619 TI - Electrotactins: a class of adhesion proteins with conserved electrostatic and structural motifs. AB - The concept of an electrostatic motif on the surface of biological macromolecules as a definite topographical pattern of electrostatic potentials in three dimensional space, provides a powerful tool for identification of functionally important regions on the surface of structurally related macromolecules. Using this approach, we identify a functional region common to cholinesterases (ChEs) and to a set of neural cell-adhesion proteins that have been suggested to be structurally related to cholinesterases due to their high sequence similarity, but lacking the key catalytically active serine. Quantitative analysis of the electrostatic surface potential in the area surrounding the entrance to the active site of acetylcholinesterase, and in the analogous zone for the ChE-like domain of the adhesion proteins reveals very good correlation. These findings, examined in the context of previous evidence involving this same region in a possible cell-recognition function for ChEs, leads us to define a class of adhesion proteins which we have named 'electrotactins'. PMID- 9725622 TI - Comparison of the specificity of homo- and heterodimeric linked HIV-1 and HIV-2 proteinase dimers. AB - The specificity of linked homo- and heterodimeric HIV-1 and HIV-2 proteinases was characterized by using oligopeptide substrates. For two substrates the k(cat)/Km values for the heterodimers were the mean values for those of the homodimers, suggesting that these substrates could productively bind into the heterodimers in both directions. However, for two other substrates the k(cat)/Km values for the heterodimers were higher than those of the homodimers, suggesting that these substrates could productively bind into the enzymes in a preferable direction. However, the mode of binding does not seem to depend on the sequential position of the subunits. The studied linked homo- and heterodimers may represent intermediate stages in the evolution of bilobal aspartic proteinases. As divergence in sequence of the two halves of such a proteinase increases, the possibility of bidirectional binding is likely lost at the expense of the optimized side-chain subsite interactions. The differences in observed and calculated k(cat)/Km values revealed dependence of the substrate specificity at one subsite of the enzyme from the next residue in sequence of substrate. These findings were also supported by molecular modeling studies. PMID- 9725621 TI - Estimates of relative binding free energies for HIV protease inhibitors using different levels of approximations. AB - Although the free energy perturbation approach is a rigorous method for estimating the relative binding free energy between an enzyme and its inhibitors, it is computationally expensive. This paper examines the accuracy at different levels of approximations, following the series expansion of free energy derived by Aqvist et al. Level-0 calculates only the enzyme-inhibitor interaction energy at the minimum energy configuration without solvent. In Level-0MD, the inhibitor configurations are sampled by molecular dynamics. These levels assume that the second- and higher order terms in the series expansion can be neglected and that the interaction energies in the bound and unbound states are equal. Level-1 does not assume equal interaction energies in the bound and unbound states. Level-1S includes the solvent contribution but both enzyme and inhibitor are fixed. In Level-1SMD, the inhibitor configurations are sampled by molecular dynamics. Level 2SMD retains the second-order term. We chose seven HIV-1 protease inhibitors for study: A77003, A76889, A76928, A78791, A74704, JG365 and MVT101. Level-0 and Level-0MD were found to give essentially the same relative interaction energies by using the AMBER force field, suggesting that fixing atomic positions may be a good approximation in some cases. However, as expected, Level-0 or Level-0MD gave poor predictions for the relative binding free energies between hydrophobic inhibitors (e.g. A77003) and more hydrophilic inhibitors (e.g. JG365). Level-1SMD produced a much better correlation between calculated and experimental results. Inclusion of the second-order term did not improve the accuracy. PMID- 9725623 TI - Probing the stabilizing role of C-terminal residues in trimethylamine dehydrogenase. AB - In trimethylamine dehydrogenase, a homodimeric iron-sulfur flavoprotein, the C terminal 17 residues of each subunit (residues 713-729) embrace residues on the other subunit. The role of this unusual mode of interaction at the subunit interface was probed by isolating three mutant forms of trimethylamine dehydrogenase in which the C-terminus of the enzyme was deleted by five residues [delta(725-729], 10 residues [delta(720-729)] and 17 residues [delta(713-729)]. The solution properties and conformational states of the three mutant enzymes were investigated using optical, fluorescence and circular dichroism spectroscopies, ANS binding and a novel and conformationally sensitive hydrodynamic method. The data reveal that sequential deletion of the C-terminus of trimethylamine dehydrogenase does not affect significantly dimer stability or the overall structural integrity of the enzyme. However, deletion of the C terminus severely compromises, but does not abolish, the ability of the enzyme to become covalently coupled with the redox cofactor FMN in the active site, located over 20 A from the C-terminus. Hydrodynamic studies reveal minor conformational changes in the deletion mutants that lead to a more compact enzyme structure. These conformational changes are probably transmitted to the active site via altering the interaction of the C-terminus with the second helix in the beta/alpha barrel of trimethylamine dehydrogenase, leading to poor flavinylation during the folding of the enzyme and assembly with FMN. PMID- 9725624 TI - A study of conserved in-loop and out-of-loop glycine residues in the large subunit of ribulose bisphosphate carboxylase/oxygenase by directed mutagenesis. AB - The replacement of all 22 completely conserved glycine residues in the large subunit of ribulose bisphosphate carboxylase/oxygenase from Anacystis nidulans by directed mutagenesis is described. In each beta/alpha barrel of the large subunit there are 12 completely conserved glycines in six of eight loops at the C-termini of eight beta-strands and four in loops at N-terminal ends of the beta-strands. Two completely conserved glycines are also in each beta/alpha barrel backbone and four more are in a large N-terminal portion preceding the barrel in a given L subunit. Substitution of glycines in loops that are C-terminal to beta-strands by proline was more deleterious to carboxylase activity than that by alanine supporting the postulates that these loops contribute to catalysis and substrate binding and that in some cases the glycines may serve as hinges enabling movement of the loops. In contrast, substitution of glycines at the N-terminal ends of beta-strands in the beta/alpha barrel more often led to failure to detect L subunits or their assembly into L8S8 complex. Substitution of these and the other conserved glycines by proline was more deleterious to carboxylase activity than by alanine in enzymes that assembled. PMID- 9725625 TI - Random mutagenesis on the Pseudomonas lipase activator protein, LipB: exploring amino acid residues required for its function. AB - LipB, lipase activator protein from Pseudomonas aeruginosa TE3285, specifically recovers the enzymatic activity of denatured inactive lipase. To find important amino acid residues of LipB in this reactivation, random mutagenesis using error prone PCR was performed on a gene encoding the functional region of LipB. The resultant DNA library was introduced into the lipase expression system using Escherichia coli, and LipB mutants lacking lipase activity were selected by two screening procedures. First, on agar plates containing tributyrin as a substrate for lipase, single colonies lacking active lipase secretion were selected as clones missing the active LipB. Second, to exclude nonsense and frameshift mutants, the molecular size of LipB in the given clones was confirmed by Western blotting. From the selected mutants, of which multiple residues are replaced, five single-residue substituted mutants were newly prepared. Consequently, Y99C, Y99H, S102R and R115C mutants formed no detectable complex with the lipase and lost the in vitro reactivation activity. In the case of Y99C and R115C, their single cysteine residue formed the intermolecular disulfide bridge. Thus, Tyr99 and Arg115 are likely to exist on the molecular surface of LipB, and are candidates for residues that make direct interaction with the denatured lipase in the reactivation process. PMID- 9725626 TI - Modification of a receptor-binding surface of epidermal growth factor (EGF): analogs with enhanced receptor affinity at low pH or at neutrality. AB - Six mutants of human epidermal growth factor (EGF), which carry single point substitutions within a surface patch proposed to juxtapose the bound receptor, were prepared and characterized for receptor affinity and mitogenicity. Receptor affinities relative to EGF are G12Q > H16D > Y13W > Q43A approximately = H16A approximately = EGF >> L15A. Notably, the reduced receptor affinity of mutant L15A indicates that Leu15 probably contributes substantially to receptor binding whereas unaltered receptor affinities observed for analogs H16A and Q43A indicate that neither His16 nor Gln43 contributes significantly to this interaction. On the other hand, the observed enhanced receptor affinities of analogs G12Q, Y13W and H16D highlight surface loci where additional productive receptor-binding contacts can be introduced. Interestingly, at acidic pH analog H16A reveals substantially greater receptor affinity than that of EGF, a property which may offer enhanced therapeutic utility in acidic environments in vivo. PMID- 9725627 TI - Spectroscopic study of Y210C lambda-repressor: implications for cooperative interaction. AB - A non-cooperative mutant of lambda-repressor, Y210C, has been purified and characterized. The mutant protein does not show any evidence of cooperative interaction as judged by difference near-UV circular dichroism spectra of DNA. The mutant protein also shows much weaker self-assembly as revealed by fluorescence anisotropy measurement. The far-UV circular dichroism spectrum of the protein shows a modest but significant reduction in the 220 nm range, suggesting a structural change. The Lehrer plot of acrylamide quenching of Y210C repressor at a predominantly dimeric concentration (0.5 microM) is almost identical with that of the wild-type protein at the same concentration. Transmission of operator-induced conformational change is also preserved in the mutant protein. Like that of the wild-type protein, cysteines of the mutant protein are unreactive to sulfhydryl reagents under native conditions. Most importantly, C210 is unreactive to sulfhydryl reagents under native conditions. This fact, coupled with the structural change observed in the far-UV CD spectra, suggests that C210 is located at the interior of the protein and exerts its effect indirectly on cooperative contact probably through destabilization of a reverse turn, of which it is an important part. PMID- 9725628 TI - Selection of cadmium specific hexapeptides and their expression as OmpA fusion proteins in Escherichia coli. AB - In searching for novel peptides with affinity for cadmium, the phage display technique was utilized. In the selection procedure, cadmium ions were immobilized on a metal chelating Sepharose gel. The peptides selected from a hexapeptide library showed no homology to naturally occurring metallothioneins. From the phage clones selected in the biopanning process, phages with affinity for Cd-109 in free solution were identified. The peptide His-Ser-Gln-Lys-Val-Phe, which was found to exhibit the strongest relative affinity for Cd-109, was cloned into Escherichia coli as a fusion to the cell surface exposed area of the outer membrane protein OmpA. Escherichia coli cells expressing this peptide showed increased survival in growth media containing up to 1.2 mM CdCl2 when compared with cells not expressing this peptide on their surface. PMID- 9725629 TI - High level expression and secretion of Fc-X fusion proteins in mammalian cells. AB - We have developed a general expression system that enhances the production and secretion of proteins in mammalian cells. The protein of interest is expressed as a fusion to a signal peptide and the Fc fragment of immunoglobulin as the N terminal fusion partner, which can direct the cellular processes into expressing and secreting high levels of many different types of proteins. These include secretory proteins, enzymes and soluble domains of membrane proteins, as well as nuclear and regulatory proteins. Typical expression levels of these proteins from stable cell lines ranged from several to 100 microg/ml in conditioned media. The Fc domain helps to solubilize hydrophobic proteins and provides a handle for easy detection and purification of the fusion proteins; and it can be cleaved off by treatment with protease if desired. PMID- 9725630 TI - Selection of anthropometric indicators for classification of abdominal fatness--a critical review. AB - In the literature, a variety of anthropometric indicators for abdominal obesity have been suggested. The criteria for their selection vary, and they have been justified mainly on the basis of being correlated with other risk factors, with morbidity and mortality, or to be predictors of the amount of visceral fat. Many of the studies, however, suffer from methodological limitations: they are based on a small number of subjects, often derived from cross-sectional data, based on indirect measurement of risk, or the indicators are complicated to interpret biologically or difficult to use in a public health context. The literature lacks a systematic evaluation of the proposed indicators taking into account possible differences between genders, age categories and ethnic groups and different diseases and mortality. Similar considerations relate to the cut-off points based on the indicators of abdominal obesity. The suggested cut-off points for waist hip ratio have been based on rather arbitrary criteria, and the studies where cut off points for waist circumference have been suggested have methodological shortcomings as well, such as being based on cross-sectional data and arbitrary cut-off points for other variables. It is also a reason for concern that so far all suggested cut-off points for abdominal obesity have been based on results obtained in Caucasian populations. Moreover, they are based on assessment of risk and their appropriateness in the use of intervention has not been evaluated. Therefore, no consensus about the appropriateness of the different cut-off points has been reached. We conclude that there is an apparent lack of consistency in the field and therefore a more scientifically and theoretically solid basis for the selection and use of anthropometric indicators of abdominal obesity and cut off points based on them should be a high priority in this research field in the near future. PMID- 9725631 TI - Short term response of circulating leptin to feeding and fasting in man: influence of circadian cycle. AB - OBJECTIVE: To investigate whether acute feeding induces changes in circulating leptin levels in humans and whether these changes vary according to nycthemeral cycle. METHODS: First experiment. Eighteen male subjects were given a fatty meal at 08.00 h. Blood sampling was performed for 10 h following this meal. Second experiment. Thirteen male subjects were given either a mixed meal or remained fasting either at night (starting at 01.00 h) or during the day (starting at 13.00 h). Blood samples were drawn every hour for a period of 8 h. RESULTS: First experiment. Serum leptin levels increased progressively from a mean (s.d.) baseline of 3.8 +/- 2.2 ng/ml to a value of 4.5 +/- 2.7 ng/ml (P < 0.01) 8 h after the fatty meal. Second experiment. During the day, serum leptin levels increased progressively from 2.65 +/- 1.7 to 3.34 +/- 2.2 ng/ml (P < 0.001) 6 h after the test-meal and decreased from 2.68 +/- 1.5 to 1.9 +/- 1.1 ng/ml (P < 0.001) 8 h after the beginning of the fasting experiment. Similar results were obtained at night. No statistically significant differences in leptin levels were observed between day and night sessions in response to feeding (mean area under the curve: 3.0 +/- 4.1 vs 4.1 +/- 4.1 ng/ml) and fasting (-2.9 +/- 2.2 vs -1.5 +/ 2.2 ng/ml). CONCLUSION: In two independent experiments, human serum leptin levels increase following food intake. This response is not influenced by nycthemeral cycle. PMID- 9725632 TI - 3-Hydroxybutyrate inhibits noradrenaline-induced thermogenesis in lean but not in obese Zucker rats. AB - OBJECTIVE: To determine the effect of 3-hydroxybutyrate (3OHB) on the thermogenic response to noradrenaline (NA) in lean and genetically obese Zucker fa/fa rats. DESIGN: Rats were infused with 18.7 nmol x kg(-1) x min(-1) of NA, supplemented, for 15 min, with 66.7 micromol x kg(-1) x min(-1) of R-3-hydroxybutyrate (3OHB). SUBJECTS: Pentobarbital-anaesthetized lean and obese Zucker rats. MEASUREMENTS: Aortic and interscapular brown adipose tissue (BAT) temperature; plasma NA, 3OHB, glucose and insulin levels during infusion. RESULTS: The NA-induced increase in aortic and BAT temperature was more marked in lean than in obese rats. In lean rats, the rise was arrested by 3OHB; but in obese rats 3OHB had no effect. Infusion of saline, glucose or 3OHB in the absence of NA did not induce changes in either temperature. NA infusion resulted in a rapid increase in plasma NA to 45-50 nM in both groups; this plateau was maintained for up to 60 min. The presence of 3OHB decreased the plasma NA of lean rats, but did not affect the plasma NA of the obese rats. Blood 3OHB rose to 1.2 mM during 3OHB infusion in both groups, and decreased on cessation of infusion. Blood glucose levels increased with NA infusion in both groups; the presence of high 3OHB levels decreased glucose levels only in lean rats. CONCLUSION: The changes in NA levels induced by 3OHB may help explain the effects observed on temperature and glucose. The defective thermogenic system of obese rats cannot be modulated by 3OHB, unlike thermogenesis in lean rats, on which 3OHB has a marked effect. PMID- 9725633 TI - Heart rate and blood pressure variability in obese normotensive subjects. AB - OBJECTIVE: To assess autonomic modulation of cardiovascular activity in massively obese subjects. DESIGN: Cross-sectional clinical study. SUBJECTS: 43 age-matched normotensive subjects: 15 moderately obese (body mass index (BMI) < 40); 14 massively obese (BMI > 40) and 14 nonobese controls (BMI < 26). MEASUREMENTS: Using power spectral analysis, heart rate and arterial pressure variability were determined at rest and after sympathetic stress (tilt). Two spectral components were analysed: a low-frequency (LF) component at around 0.1 Hz, predominantly reflecting sympathetic modulation and a high-frequency (HF) component at around 0.26 Hz, reflecting parasympathetic modulation. RESULTS: Spectral data for heart rate showed that the massively obese subjects had lower LF [mean +/- s.e.m.] normalized units (NUs) at rest (35.1 +/- 0.9) and after tilt (56.1 +/- 2.1), than the moderately obese subjects (LF NUs at rest 53.9 +/- 4.2, P < 0.001; LF NUs tilt: 66.8 +/- 5.6, P < 0.001) and nonobese control subjects (LF NUs at rest, 56.6 +/- 3.0, P < 0.001); (LF NUs tilt: 81.7 +/- 1.7, P < 0.001). Data for systolic arterial pressure variability measured at rest exhibited the inverse pattern, the massively obese group having higher mean LF values (LF mm Hg2 rest: 15.0 +/- 1.4; LF mm Hg2 tilt: 15.7 +/- 1.5) than the moderately obese group (LF mm Hg2 rest 3.2 +/- 0.7, P < 0.001; LF mm Hg2 tilt: 7.2 +/- 2.0, P < 0.001) and than the nonobese control subjects (LF mm Hg2 rest 3.5 +/- 0.5, LF mm Hg2 tilt 8.5 +/- 0.8, P < 0.001). Regression detected a significant association between BMI and LF of systolic pressure (beta = 0.364; P = 0.0007), In LF of heart rate (beta = -5.555; P = 0.00001) and very low frequency (VLF) of diastolic pressure (beta = -3.305; P = 0.0020). CONCLUSION: Obesity seems to increase sympathetic modulation of arterial pressure, but diminishes modulation of heart rate. Because our obese subjects had high plasma noradrenaline levels, their low LF power of heart rate could reflect diminished adrenoceptor responsiveness. PMID- 9725634 TI - Effect of exercise and tepary bean type diet on body composition and fat accretion in obese Zucker rats. AB - OBJECTIVE: The effectiveness of a tepary bean high fat type diet, compared to a purified type high fat diet and exercise, on body composition in fatty Zucker rats was determined. SUBJECTS AND DESIGN: Approximately 6-week-old female fa/fa Zucker rats were divided into four groups of 10 rats each: TE, fed the tepary bean type diet and exercised; TN, fed the tepary bean type diet and not exercised; CE, fed the purified type control diet and exercised; CN, fed the purified type control diet and not exercised. The exercise modality was treadmill running and the experiment lasted 13 weeks. MEASUREMENTS: Body weight, cumulative food intake, body composition, weights of adipose tissues and liver, heart and gastrocnemius muscle. RESULTS: At the end of the 13 week experiment, TE rats weighed 511 +/- 22 g and were significantly lighter than TN, 588 +/- 15 g; CE, 606 +/- 22 g; and CN, 660 +/- 27 g. All are means +/- s.e.m. The carcass of CN rats had 58, 20 and 13% more fat than TE, TN and CE rats, respectively; P < 0.01. Lean body mass was the same for all the groups of rats and ranged from means of 216-228 g. However, TE rats had significantly more fat free dry mass (FFDM) than CN rats; 68 +/- 4 vs 58 +/- 2 (means +/- s.e.m.) and tended to have more FFDM than TN and CE rats. Inguinal fat depots weighed 20-30% less in T than in C rats (diet comparisons) and also 20-30% less in E than in N rats (exercise comparisons). Perirenal/retroperitoneal fat depots weighed 25% less in TN than in CN rats and 38% less in TE than in CE rats. Exercise did not reduce perirenal/retroperitoneal fat depot weights. Parametrial fat depot weights were not influenced by diet or exercise. CONCLUSIONS: In diets which provided 37% of the energy from fat, the incorporation of tepary beans attenuated weight gain, and subcutaneous and visceral fat gain compared to a purified type diet. Exercised rats gained less weight and subcutaneous, but not visceral fat, than non-exercised rats. PMID- 9725635 TI - Influence of diet, physical activity and parents' obesity on children's adiposity: a four-year longitudinal study. AB - OBJECTIVE: To assess the relationships between diet, body composition, physical activity, parents' obesity and adiposity in children at the age of 8 y and four years later. STUDY DESIGN: Prospective observational study of anthropometric measures initiated in 1992, follow-up examination in 1996. METHODS: 112 prepubertal (age: 8.6 +/- 1.0 y) children were studied. Energy and nutrient intakes were assessed by diet history, body composition by anthropometry and physical activity, by a questionnaire. Obesity was defined as relative body mass index (BMI) (rel BMI) > 120%, where rel BMI = (BMI/BMI at 50th centile for age and gender) x 100. RESULTS: Prevalence of obesity was not statistically different at baseline (22.3%) than four years later (19.8%): rel BMI at the age of 8 y was positively self-related with rel BMI at the age of 12 y (r = 0.73, P < 0.001). After four years, eight (32%) obese children became non obese and five (6%) non obese children became obese. Multiple regression analysis (stepwise procedure) revealed that, in the final equation, the mother's BMI and TV viewing (independent variables) accounted for 17% of the children's rel BMI variance at the age of 8 y (R = 0.42, P < 0.001) while the parents' BMIs accounted for 13.5% of the children's rel BMI variance at the age of 12 y (R = 0.37, P < 0.001). Other variables such as total energy intake, nutrient intake percentage and amount of physical activity, were all rejected. An autoregressive unbalanced measures model regression analysis recognised the mother's and father's BMIs as the only variables able to predict rel BMI in the children (mother's BMI coeff. 2.53 (s.e.m. 0.26), P < 0.0001; father's BMI coeff. 2.07 (s.e.m. 0.23), P < 0.0001). A multivariate logistic regression analysis was also performed. The children who participated in the follow-up, were divided into two groups based on the positive or negative change in the rel BMI between final and baseline measurements. Of all the variables considered, only rel BMI at baseline was selected in the final equation. Other variables such as age, gender, energy and nutrient intake, TV viewing and amount of physical activity, as well as the parents' BMI, were all removed. CONCLUSIONS: The parents' obesity was the main risk factor for obesity in this group of children. Sedentary behaviour (TV viewing) was independently associated with overweight at the age of 8 y. Physical activity and energy and nutrient intakes did not significantly affect the change in rel BMI over the four-year period when the parents' obesity was taken into account. PMID- 9725636 TI - Leptin: its pharmacokinetics and tissue distribution. AB - OBJECTIVE: The pharmacokinetics and tissue distribution of leptin in rats was investigated. DESIGN: A catheter was inserted in the right jugular vein of rats on the day prior to experiment. The next day, blood was sampled and then a tracer dose of radioiodinated hormone was administered via the catheter. Thereafter, small (200 microl) samples of blood were taken at regular intervals. Two experiments were conducted over different sampling times. TCA precipitated radioactivity was counted in samples of plasma and tissues. Pharmacokinetic parameters were calculated after fitting a bi-exponential equation describing a two-pool model of plasma leptin distribution. Selected time-point plasma samples were fractioned using size exclusion chromatography and the leptin distribution determined. RESULTS: The two pool model described the pharmacokinetics of leptin in two forms: an initial fast decaying pool (t(1/2) = 3.4 min) and a slower decaying pool (t(1/2) = 71 min) with an overall clearance rate of 6.16 ml/min/kg. Size exclusion chromatography showed a persistent peak (all time-points tested) of 125I-leptin corresponding to the plasma albumin peak. The size of the free 125I-leptin peak became diminished or absent in later time-point plasma samples. Tissue distribution of leptin at 60 min and 180 min time-points showed that the small intestine contained the highest concentration of leptin, almost four times the level found in kidneys, liver, stomach and lungs. 125I-leptin was least abundant in skin, muscle, heart, caecum and brain. CONCLUSION: The pharmacokinetics of leptin are affected by three important factors: 1) its ability to bind to a plasma carrier molecule which increases its half-life; 2) its association with abundant peripheral tissue binding sites which creates an additional pool of leptin and 3) the rate of synthesis of leptin which may be less important than originally believed as the prolonged half-life and the additional pool of tissue binding sites are important factors in determining its plasma concentration. PMID- 9725637 TI - Association between obesity and high blood pressure: reporting bias related to gender and age. AB - OBJECTIVE: To examine the validity of self-reported information on obesity and high blood pressure (HBP) in relation to gender and age, and to explore the impacts of their misclassification on the association between obesity and HBP. DESIGN: Community based cross-sectional study. SUBJECTS: 1791 adult subjects living in Humboldt, Saskatchewan, Canada. MEASUREMENTS: Objectively measured HBP was positive if systolic blood pressure (BP) was > or = 140 mm Hg, diastolic BP was > or = 90 mm Hg or the subject was currently using antihypertensive medication. Self-reported HBP was positive if the subjects gave an affirmative response to the question: 'Has a doctor ever said you had high blood pressure?' Body mass index (BMI) was calculated as weight (kg)/height (m)2. Obesity was defined as a BMI > 27 kg/m2. Measured obesity and reported obesity were based on measured and self-reported information on height and weight, respectively. RESULTS: The sensitivity of self-reported HBP was low, and was lower for men than for women, and for younger subjects than for older subjects. The specificity was similar for both genders. Obese individuals had higher sensitivity and lower specificity than non-obese individuals. The differential misclassification of self-reported HBP caused a bias away from the null when the relative risk for HBP in relation to obesity was estimated. CONCLUSIONS: As a result of the gender- and age-related misclassification of self-reported HBP, the modification role of gender and age on the association between obesity and HBP could be altered. The bias caused by self-reported obesity was relatively small and was either toward or away from the null. PMID- 9725638 TI - The Eating Inventory in obese women: clinical correlates and relationship to weight loss. AB - OBJECTIVES: Describe the physical and psychological correlates of the Eating Inventory (EI) (also known as the Three-Factor Eating Questionnaire) factors in an obese sample, and determine the relationship between the three EI factors and weight loss. DESIGN: Consecutive series of obese women enrolled between 1987 and 1996 in clinical trials of weight loss treatments. PARTICIPANTS: 223 obese women with a weight of 100.7 +/- 15.5 kg, an age of 41.4 +/- 8.8 y and a body mass index (BMI) of 37.2 +/- 5.6 kg/m2. MEASURES: The EI and a variety of physical (weight, body composition and resting energy expenditure) and psychological (mood and binge eating) measures were assessed before and after 5-6 months of treatment. RESULTS: Before treatment, higher restraint scores were associated with lower body weights (P = 0.02), while higher disinhibition scores were associated with greater binge eating severity (P<0.0001). Weight loss treatment was associated with significant increases in restraint and decreases in disinhibition and hunger (all Ps<0.0001). Greater increases in restraint during treatment were associated with larger weight losses (P<0.0001). CONCLUSIONS: The three factors of the EI showed clinical utility in a sample of women receiving treatment for obesity. PMID- 9725639 TI - Is body composition important in young people's weight management decision making? AB - AIM: Young people are often seen as an important target for prevention of overweight, but we know little about the factors which are important for their weight management decisions. This study aimed to evaluate the extent to which elements of body composition and dimensions were implicated in their decisions to change their weight. PARTICIPANTS: Participants were 116 male and 126 female volunteers from a tertiary college in the south-west of England with a mean age of 17.90 (s.d. = 1.90) y. MEASURES: Body composition measures included height (m), weight (kg), waist and hip circumferences (mm), shoulder and hip girths (mm) and skinfold thicknesses (triceps, calf and subscapular, mm). Students also provided self-reported information on dietary practices they had undertaken in the last year. RESULTS: More females reported attempting weight loss in the last year compared to males (51.5 vs 17.6%), although more males reported attempting weight gain (19.3 vs 2.3%). Reported dietary strategies for both males and females centred around eating less fatty foods, eating less than usual and exercising more. However, a significant proportion of females also reported excessive strategies, such as self-induced vomiting and regular crash dieting. Results indicated that females who desired to lose weight were significantly (P < 0.05) heavier, and bigger in terms of circumferences and skeletal breadths, compared to females who did not wish to change their weight. For males, the 'weight loss' group were significantly (P < 0.05) bigger, heavier and fatter compared to the 'no change' and 'weight gain' group. For females only, the previously significant main effect for body dimensions across weight management groups (Pillais F(14,188) = 3.81, P < 0.001) was extinguished when controlling for bone dimensions (Pillais F(12,198) = 1.68, P = 0.074). CONCLUSION: These results indicate that frame size, particularly in the lower trunk, is a more important predictor of female weight management decision-making, than their levels of fatness. For males, fatness appears to drive their weight management decision-making processes to a greater extent. PMID- 9725640 TI - Resting metabolic rate in preadolescent girls at high risk of obesity. AB - OBJECTIVE: To investigate whether children at high risk of obesity have a reduced resting metabolic rate (RMR). DESIGN: Cross-sectional study. SUBJECTS: 93 healthy girls (age: 8-12 y) were allocated to one of four groups, according to the subjects' and their parents' weight status: group 1, overweight children with both parents overweight (OB/OB2; n = 17); group 2, normal weight children with both parents overweight (N/OB2; n = 28); group 3, overweight children of discordant parents (OB/OB1; n = 21) and group 4, normal weight children with both parents normal weight (N/OB0; n = 27). MEASUREMENTS: Weight, height, fat-free mass (FFM) and fat mass (FM) by bioelectrical impedance analysis (BIA), RMR (indirect calorimetry) for the duration of 25 min. RESULTS: Controlling for disparities in body composition, age and physical maturity, significant differences were found in adjusted group means of RMR (OB/OB2 1181 kcal/d; N/OB2 1276 kcal/d; OB/OB1 1234 kcal/d; N/OB0 1199 kcal/d; P < 0.02) with the OB/OB2 girls showing the lowest energy expenditure. CONCLUSION: We found evidence that preadolescent girls at risk of obesity, are not generally predisposed to a higher body weight, because of a greater metabolic efficiency. In fact, our data show that more emphasis should be laid on defining different subgroups of both overweight and normal weight subjects in studies investigating metabolic rate. PMID- 9725641 TI - Inhibition of insulin release by enterostatin. AB - OBJECTIVE: These studies were designed to investigate the mechanism through which enterostatin inhibits insulin secretion from pancreatic islets. DESIGN: A static islet incubation method was used to study the effects of enterostatin on insulin secretion induced by various secretagogues and to investigate the effect of calcium ions and 8-Br-cyclic AMP on the response to enterostatin. Measurements of islet cAMP concentrations in response to enterostatin were also made. RESULTS: Enterostatin (10(-9) to 10(-5) M) inhibited insulin secretion from islets incubated in the presence of 16.7 mM glucose in a dose-dependent manner. Enterostatin also inhibited insulin secretion stimulated by glybenclamide (5.0 and 10 microM), phorbol 12-myristate-13-acetate (TPA) (50 and 100 nM), and the kappa-opioid agonist U50,488 (100 nM). The inhibitory effect of enterostatin on TPA-induced insulin secretion was attenuated but still remained in the absence of extracellular Ca2+. The enterostatin inhibition of insulin secretion was blocked by 8-Br-cAMP (1 mM) independent of extracellular Ca2+. Enterostatin reduced the increase in intracellular cyclic AMP (cAMP) content produced by U50,488 (100 nM) and the changes in cAMP content were parallel with changes in insulin release. CONCLUSION: Enterostatin may suppress insulin secretion through the reduction of cAMP, but other mechanisms may also be possible. PMID- 9725642 TI - Peripheral fat metabolism during infusion of an exogenous triacylglycerol emulsion. AB - OBJECTIVE: To test the hypothesis that intravenous infusion of lipid would bring about changes in adipose tissue metabolism, which would tend to spare net fat mobilization, and to attempt to identify the mediators of such responses. DESIGN: The triacylglycerol (TG) emulsion, Intralipid, was infused and metabolic changes in subcutaneous adipose tissue and forearm muscle were assessed by measurements of arterio-venous differences. SUBJECTS: Six normal male subjects aged 21-37 y, with body mass index (BMI) 23.0-25.9 kg/m2. RESULTS: Plasma TG and non-esterified fatty acid (NEFA) concentrations rose during infusion as expected. The rise in systemic plasma NEFA concentration occurred despite decreased NEFA release from adipose tissue. Intralipid infusion resulted in a suppression of intracellular lipolysis in adipose tissue, by mechanisms which are not clear. Plasma leptin concentrations, measured in a search for the regulator of lipolysis, showed consistent leptin release from adipose tissue which did not change significantly with time. CONCLUSION: The suppression of intracellular lipolysis in adipose tissue during Intralipid infusion is a new observation and may reflect a novel mechanism for regulation of fat storage. PMID- 9725643 TI - The effects of dietary-induced obesity on the biomechanical properties of femora in male rats. AB - OBJECTIVE: To assess the effects of diet-induced obesity (DIO), on the biomechanical and biochemical properties of the femur in mature male rats. DESIGN AND SUBJECTS: Two groups of male rats were studied. The DIO experimental group was fed a high caloric diet and a 31% sucrose solution as drinking fluid for a month, whereas the control group was fed lab chow and tap water. MEASUREMENT: Body weight; body water; lean body mass; femoral length; average cortical thickness; outer anteroposterior diameter; outer mediolateral diameter; cortex area; moment of inertia; cortical and cancellous bone hydration; tendon and muscle hydration; ash content of cortical and cancellous bone; ultimate load; deflection at ultimate load; ultimate strength; stiffness; elastic modulus and energy absorption capacity. RESULTS: 'Gainers' (final body weight in excess of three standard error of mean of the controls) were 19.1% heavier, with higher body fat, whereas body water, lean body mass, hydration of cancellous bone and ash content of cortical bone were lower, when compared to controls. Rats that failed to gain weight, despite the high caloric diet, were termed 'resisters' (weight gain less than three standard error of mean of the controls). Ultimate load, deflection at ultimate load and femoral energy absorption capacity were significantly higher in the experimental group when compared to the controls. However, no differences were found among the groups with respect to ultimate stress and stiffness. CONCLUSION: The weight gain produced by DIO may lead to bone adaptation and improved biomechanics. PMID- 9725644 TI - Serum leptin, food intake and preferences for sugar and fat in obese women. AB - OBJECTIVE: To examine the association of leptin with food intake and preferences for sugar and fat in obese humans. METHOD: Food intake and preferences for sugar and fat were measured in 35 obese women by a four-day food record and three hedonic tests, respectively. RESULTS: High fasting serum leptin concentration adjusted for body fat mass and dietary underreporting was associated with low dietary energy and fat intakes. In addition, trends towards lower preference for chocolate as well as for the taste of high-fat, low-sugar mixture were observed in those with higher leptin concentration. CONCLUSION: High serum leptin concentration could be associated in obese women with lower dietary energy and fat intakes, and possibly with the lower preference for fat. These findings need to be verified in further human studies. PMID- 9725645 TI - Serum leptin and weight loss in severely obese patients undergoing biliopancreatic diversion. AB - OBJECTIVE: To evaluate the influence of body fat and food intake on serum leptin concentration. DESIGN: Longitudinal study of a group of obese patients prior to and at, long term follow-up, after biliopancreatic diversion (BPD), when body weight was steadily reduced and food consumption was similar to or greater than preoperatively. RESULTS: In obese patients, very high serum leptin concentrations were found. Following the operation, with the body weight stable and normalized, a sharp fall of serum leptin concentration had occurred, with values returned to normal range. CONCLUSION: The changes in serum leptin concentration observed in the long term after weight loss are substantially accounted for by the loss of body fat and appear unrelated to the reduction of oral food intake. PMID- 9725646 TI - Poor systematic reviews and meta-analyses may be misleading. PMID- 9725647 TI - TRI2SOLID: an application of reverse engineering methods to the creation of CAD models of bone segments. AB - For many biomechanical engineering activities it would be useful to have the three dimensional (3D) geometry of bone segments available in form of vectorial models within computer aided design (CAD) environments. In this paper a new method for the semi-automatic conversion of a stack of CT images of a femur into a CAD solid model is described. This method is relatively simple, accurate, and requires only a 3D CAD plus a few additional programs available in the public domain. The proposed method was used to convert the CT scan data set of a human femur into a valid CAD model; the resulting solid was two times more accurate than that obtained using the commonly used procedure based on 2D segmentation. PMID- 9725648 TI - Classification and identification of proteins by means of common and specific amino acid n-tuples in unaligned sequences. AB - Unaligned amino acid sequences can be characterized by their composition of amino acid n-tuples (i.e. doublets, triplets, quadruplets, etc.). In this study we investigated the performance of two statistics, termed commonality and specificity, that are derived from n-tuple counts using a set of G-protein coupled receptor (GPCR) sequences. The commonality of a tuple is defined as its relative occurrence in the sequences that belong to a given GPCR subtype. The specificity of a tuple is derived from its relative occurrence in the sequences of a given GPCR subtype and from its relative non-occurrence in the sequences that do not belong to this subtype. A graphical presentation, termed 'polygram', is described for the visualization of common and specific tuples. The method can be applied to the classification of unknown GPCR sequences. It can also be applied to the identification of fragments of GPCRs, such as may occur in chimeric receptors. The method is generally applicable to other protein families and other types of coding. PMID- 9725649 TI - Modeling and forecasting monthly patient volume at a primary health care clinic using univariate time-series analysis. AB - Two univariate time-series analysis methods have been used to model and forecast the monthly patient volume at the family and community medicine primary health care clinic of King Faisal University, Al-Khobar, Saudi Arabia. Models were based on nine years of data and forecasts made for 2 years. The optimum ARIMA model selected is an autoregressive model of the fourth order operating on the data after differencing twice at the nonseasonal level and once at the seasonal level. It gives mean and maximum absolute percentage errors of 1.86 and 4.23%, respectively, over the forecasting interval. A much simpler method based on extrapolating the growth curve of the annual means of the patient volume using a polynomial fit gives the better figures of 0.55 and 1.17%, respectively. This is due to the fairly regular nature of the data and the lack of strong random components that require ARIMA processes for modeling. PMID- 9725650 TI - Magnetic resonance image processing and structured grid generation of a human abdominal bifurcation. AB - Magnetic resonance angiography (MRA) offers a non-invasive approach to the acquisition of anatomically accurate human arterial structure. Combining the latest computational fluid dynamics (CFD) techniques with clinical data from MRA, the detailed haemodynamics information in the human circulation system can be obtained. In this paper, a novel computer method is presented, which generates automatically a computational grid for a human abdominal bifurcation from a set of conventional MRA images. The method covers the complete sequence from MR image segmentation, 3-D model construction, grid generation, to grid quality evaluation. Results demonstrate that the computer program developed is capable of generating a good quality grid for human arterial bifurcations from MRA images with minimum user input. The resultant grid can be used directly for further computer simulation of the flow. PMID- 9725651 TI - Development in a Windows environment of a radiation treatment planning system for personal computers. AB - A new personal computer (PC) radiotherapy treatment planning system (RTPS) is presented. The PC-based RTPS is designed to run in the Microsoft Windows 3.11 environment (and later versions), for computers equipped with 486 or Pentium processors. The algorithm used by the new PC-based program for dose calculation belongs to the 'radiological pathlength' category and it was previously implemented on VAX 711 computers at Memorial Sloan-Kettering Cancer Center (MSKCC) in New York, NY, within EXTREP-III RTPS. The EXTREP-III program is a two dimensional RTPS (with restricted three-dimensional capabilities), developed and used in clinical practice at MSKCC during the 1980s. The PC-based program is implemented in the Visual Basic (version 3.0) language and supports features commonly available in most photon-mode RTPSs: dose calculation for fixed, isocentric and rotational irradiation techniques, dose corrections for both internal inhomogeneities and external inhomogeneities (boluses and compensators), association of machine-specific beams with various wedges and blocks, etc. The graphic interface of the PC-based RTPS is completely new and is designed to meet the requirements of fast and accurate planning. The user interface consists of an event-oriented button-based console which allows users to perform planning and to have isodose charts overlaid on patient computed tomography images initially loaded in the program. The PC-based RTPS tests, performed in order to assess its accuracy and speed of computation, show good results. The acceptable computation times obtained, the good accuracy in dose computation and the user-friendly interface of the program are sufficient reasons to consider the PC-based RTPS a good quality-price ratio tool for radiation treatment planning in cancer therapy. PMID- 9725652 TI - The Multi Unit Activity analyzer: a Windows based hardware-software system for low cost, high speed analog to digital data conversion, data acquisition and window discrimination. AB - The Multi Unit Activity analyzer is a hardware-software package for multi purpose, two-channel data acquisition, with a computer dependent maximal digitizing frequency selectable from 1 to 27,000 s(-1) on both channels simultaneously. The hardware is connected to an IBM compatible PC through one of the serial ports (standard RS 232 interface). Software was developed to view digitized signals and record or read them on or from the harddisk. The program can also perform amplitude based window discrimination on the raw signal, on-line or during replay. The system is used for recording and analyzing multi unit activity from neuronal tissue in our electrophysiology lab but it can be applied in a variety of other settings. Basic programming routines are available that allow customized data acquisition. PMID- 9725653 TI - The effect of under- and overnutrition on essential fatty acid metabolism in childhood. AB - Both protein-energy malnutrition (PEM) and obesity represent major challenges for paediatric nutrition. The aim of this review is to summarise available data regarding the effect of PEM and obesity on the availability of essential- and long-chain polyunsaturated fatty acids (LC-PUFAs) in childhood. Significantly lower arachidonate (C20:4n-6, AA) values in malnourished children than in controls is a consistent finding in all studies, whereas it is controversial whether the availability of docosahexaenoate (C22: 6n-3, DHA) is also affected. We found significantly lower percentages (% wt/wt) of both AA and DHA in plasma phospholipids [AA: 7.0 (0.7) vs 8.7 (0.8); DHA: 0.90 (0.2) vs 2.6 (0.7), median (interquartile range), P < 0.001] in severely malnourished children aged 29 (7) months than in control subjects. Product/substrate ratios indicated reduced delta 5-desaturation in children with PEM. We speculate that severely malnourished children may benefit from enhanced dietary supply of both n-6 and n-3 LC-PUFAs. In obese adults AA has been reported to constitute a lower percentage of plasma phospholipid fatty acids, and AA supplementation of weight reduction diets has been suggested. In contrast, we found significantly higher plasma phospholipid AA values [12.6 (2.4) vs 8.3 (1.4), P < 0.001] in markedly obese children aged 13.8 (1.1) years than in non-obese controls. Product/substrate ratios of the delta-6 desaturase enzyme indicated enhanced conversion activity. These data suggest that obese children do not require LC-PUFA supplementation to low fat diets. The available data indicates that both PEM and obesity alter fatty acid composition of plasma and erythrocyte membrane lipids. The underlying mechanism appears to be altered activity of the bioconversion of essential fatty acids to their LC-PUFA metabolites. PMID- 9725654 TI - By how much does fruit and vegetable consumption reduce the risk of ischaemic heart disease? AB - OBJECTIVE: To quantify the relationship between fruit and vegetable consumption and the incidence of ischaemic heart disease. DESIGN: A meta-analysis of cohort studies of the relationship between ischaemic heart disease and markers of fruit and vegetable consumption, namely dietary intake of fruit, vegetables, carotenoids, vitamin C, fruit fibre and vegetable fibre, and serum concentration of carotenoids and vitamin C, adjusted for other risk factors. MAIN OUTCOME MEASURES: Risk of ischaemic heart disease at the 90th centile of consumption relative to that at the 10th, equivalent to about a four-fold difference in fruit consumption and a doubling of vegetable consumption. RESULTS: The association with ischaemic heart disease was of similar magnitude for all six dietary markers of fruit and vegetable consumption. The median of the six estimates was that risk was 15% (range 12-19%) lower at the 90th centile of consumption than at the 10th. The estimates were generally adjusted for the possible confounding effect of other heart disease risk factors. The serum studies of vitamin C were consistent with this; those of carotenoids suggested a larger difference (43%) but were not adjusted for the important confounding effect of smoking. The substances in fruit and vegetables responsible for the protective effect on heart disease are uncertain but the effect is commensurate with the estimated protective effects of the potassium and folate in fruit and vegetables. Beta-carotene or vitamin E are not likely to be important because randomised trials of these vitamins in large doses have shown no reduction in heart disease mortality. CONCLUSIONS: The risk of ischaemic heart disease is about 15% lower at the 90th than the 1Oth centile of fruit and vegetable consumption. PMID- 9725655 TI - Validation of a short food frequency questionnaire to assess consumption of cereal foods, fruit and vegetables in Chinese Singaporeans. AB - OBJECTIVE: To assess the ability of a 16-item food frequency questionnaire (FFQ) to measure consumption of cereal foods, fruit and vegetables in Chinese Singaporeans. DESIGN: Subjects completed the questionnaire twice, at the beginning and end of a six-week period during which they also provided three 24 h diet recalls. Estimates of intake from the questionnaire were compared with those from diet recalls. SUBJECTS: Subjects were recruited from a range of occupational groups through random sampling across divisions in the headquarters of the Singapore Ministry of Health. Of the 81 subjects initially recruited, three failed to complete the diet recalls, one was excluded due to changes in diet, and seven did not return the second questionnaire. RESULTS: Mean difference in food group consumption estimated by the two methods did not differ significantly from zero for fruit (0.00 serving, s.d.=0.54, 95% CI= -0.13, +0.12, P=0.95) or vegetables (-0.05, s.d.=0.29, 95% CI= -0.12, +0.02, P=0.13). For cereal foods, the mean difference was small, but significantly different from zero only in women (-0.32 servings, s.d. = 0.92, 95% CI = -0.59, -0.06, P=0.02). At an individual level, cereal food intake as measured by the FFQ may be 37% below or 59% above the diet recall values; and values for total fruit and vegetables may be half or double the recall values. Among subjects whose intake was classified into the lowest quartiles by diet recalls, 78% and 94% respectively, fell into the lowest two questionnaire quartiles for cereal foods, and total fruit and vegetables. The ability of the questionnaire to predict those having inadequate intake based on recall data was more than 90% for the three food groups. CONCLUSION: The short questionnaire cannot replace the three-day recalls in intake assessment for individuals, but it could be used to screen for low consumers in intervention programmes, to assess mean food group intake in population groups, and to rank individuals into broad categories of food group intake. PMID- 9725656 TI - Relationship between dietary habits, age, lifestyle, and socio-economic status among adult Norwegian women. The Norwegian Women and Cancer Study. AB - OBJECTIVE: To examine how dietary intake varies with age in a nation-wide sample of adult Norwegian women, and to evaluate the impact of lifestyle and socio economic status on important dietary aspects. DESIGN: Cross-section study. SETTING AND SUBJECTS: A food frequency questionnaire was mailed to a random, nation-wide sample of 20 000 women aged 45-69y, and 9885 questionnaires were accepted for nutritional analyses. RESULTS: Dietary habits differed moderately with age. The oldest women reported a higher consumption of potatoes and fish, whereas the youngest reported more coffee, meat, and alcohol. The reported intake of fruit, vegetables, and potatoes was lower than recommended in all age groups. Older women had a slightly better distribution of energy yielding nutrients than younger women, although the median percentage of energy from fat was too high in all age groups. The median dietary fibre density of the diet was close to the recommended level in all age groups, yet lowest among the youngest women. Practising a healthy lifestyle and having a higher socio-economic status were associated with reporting a healthier diet. However, adjusting for lifestyle and socio-economic factors did not substantially alter the associations between diet and age. CONCLUSIONS: Older women tend to have a healthier diet than younger women. The relationship does not seem to be strongly confounded by lifestyle and socio-economic status, although these factors are also related to dietary habits. PMID- 9725657 TI - Skinfold thickness measurements are better predictors of body fat percentage than body mass index in male Spanish children and adolescents. AB - OBJECTIVE: To develop equations, from some simple anthropometric measurements, for the prediction of body density from underwater weighing in male spanish children and adolescents. SUBJECTS: One hundred and seventy-five males, aged 7.0 16.9 y, participated in this study, they were recruited from primary and secondary schools. MEASUREMENTS: Body weight and height and skinfold thicknesses by anthropometry, body density by underwater weighing. RESULTS: Correlations between body density and body mass index (BMI) were high until 14.0-16.9y. Correlations between body density and log sigma 4 skinfolds were higher than those with BMI at all ages. Log sigma 4 skinfolds explained between 61% (14.0 16.9 y) and 68% (11.0-13.9 y) of the body density variance. Regression equations for body density from BMI and triceps skinfold thickness explained between 51% (14.0-16.9y) and 68% (7.0-10.9 y) of the body density variance. CONCLUSIONS: The best estimators of body density in the children and adolescents studied were log sigma 4 skinfolds and a combination of BMI and triceps skinfold. PMID- 9725658 TI - Prevalence of self-reported food hypersensitivity among school children in The Netherlands. AB - OBJECTIVES: To provide national figures on the prevalence of self-reported food hypersensitivity (S-FH), and the association with socio-demographic variables and some health indicators in schoolchildren in The Netherlands. DESIGN: As part of the Child Health Monitoring System, data were collected from 4450 children, who were invited for a routine health assessment (response 97%). A questionnaire on food hypersensitivity was completed by the parents of the children in primary school and by the children in secondary school themselves. The measurements on health status were taken by the school physician or nurse during the school health assessment. SUBJECTS: Children aged 4-15 y in The Netherlands in three groups in primary school, and in the second year of secondary school. RESULTS: The prevalence of S-FH was 7.2%. Food additives and chocolate were most frequently avoided. Of the children with S-FH, 40% avoided food exclusively either on their own accord or on advice of relatives. School absence due to illness, use of medication, and medical treatment were more prevalent in children with S-FH, and their health status was more often considered moderate or poor by the school physician or nurse. CONCLUSION: Seven percent of school-aged children avoid certain types of food or ingredients because of S-FH. The prevalence of food allergy or food intolerance is probably lower, since many children with S-FH had not undergone any diagnostic tests. To prevent unnecessary food restriction, more information for parents is needed about the possible effects of food restriction on the health of their children, and they should be encouraged to seek further diagnosis. PMID- 9725659 TI - Dietary intake of cobalamin in elderly people who have abnormal serum cobalamin, methylmalonic acid and homocysteine levels. AB - OBJECTIVE: To determine if poor dietary intake can explain the cobalamin-related abnormalities often seen in the elderly. DESIGN: Prospective laboratory survey with a follow-up dietary assessment. SETTING: Social centers for the elderly and an outpatient clinic. SUBJECTS: Ninety-five free-living subjects >60y old with abnormal or suspicious findings in cobalamin-related tests and 78 subjects >60y old with normal results. INTERVENTIONS: Serum cobalamin, methylmalonic acid and homocysteine determinations to assess cobalamin status and a one year food frequency questionnaire to assess cobalamin intake. RESULTS: Only three of the 173 subjects (1.7%), one of whom had normal cobalamin status, ingested <2 microg cobalamin/d, the Recommended Daily Allowance. Sixty-nine subjects (39.9%) ingested <6 microg/d, but they did not have more abnormal serum cobalamin or metabolite values than those ingesting >6 microg. Ordering all subjects by quintiles according to cobalamin intake revealed no significant trends or differences in any of the serum values either. Moreover, arranging subjects by results of tests of cobalamin status showed that the subjects with abnormal cobalamin status did not differ in cobalamin intake from those with normal cobalamin status, although they did differ in use of supplements. Finally, cobalamin intake, with or without supplements, did not correlate with serum cobalamin or metabolite levels. The absence of any association between cobalamin status and intake contrasts sharply with the significant correlation between folate intake and folate status (P = 0.0001). CONCLUSIONS: The high frequency of mildly abnormal cobalamin status in the elderly cannot be attributed to poor intake of cobalamin. Nondietary explanations, such as malabsorption and other phenomena, must always be sought to explain mild cobalamin deficiency in the elderly. PMID- 9725660 TI - Dietary assessment in the elderly: validation of a semiquantitative food frequency questionnaire. AB - OBJECTIVE: The study was conducted to assess the relative validity of a 170-item semiquantitative food frequency questionnaire (SFFQ) adapted for use in the elderly. DESIGN AND SUBJECTS: The study was carried out in a sample of 80 men and women aged 55-75 y participating in a community based prospective cohort study in Rotterdam, The Netherlands. The two-step dietary assessment comprised a simple self-administered questionnaire (20 min) followed by a structured interview with trained dietitians (20 min) based on the completed questionnaire. Multiple food records (FR) collected over a one year period served as reference method. 24 h urine urea was used as indirect marker for protein intake. RESULTS: Compared with FR, the SFFQ generally overestimated nutrient intake as reflected by difference in means and the ratio of SFFQ to FR. Energy adjustment reduced the observed overestimation. Pearson's correlation coefficients varied from close to 0.5 to about 0.9 for crude data, and after adjustment for age, sex, total energy intake, and for within-person variability in daily intake for 0.4-0.8. Cross classification into quintiles resulted in correct classification into the same or adjacent quintile of 75.8% for crude and 76.8% for energy-adjusted data. Validation of protein intake estimated by SFFQ with protein excretion from 24h urine urea indicated overestimation of protein intake by SFFQ. Spearman correlation coefficient between protein intake estimated from urea excretion and SFFQ was 0.67. CONCLUSIONS: Adaptation of a SFFQ for use in the elderly resulted in a valid and time-efficient dietary assessment instrument. Its ability to adequately rank study subjects according to their dietary intake support its application in epidemiological studies in the elderly. PMID- 9725661 TI - Low-dose spiral computed tomography for measuring abdominal fat volume and distribution in a clinical setting. AB - OBJECTIVES: Computed tomography (CT) has been used to measure body composition, however, a technique with reduced radiation exposure has not yet been introduced. This study tested a low-dose spiral CT technique on a phantom to determine its validity and reproducibility. The method was then applied for volume and distribution measurements in patients. DESIGN: Construction and measurement of a phantom followed by measurement of patients referred to CT for clinical indications. SETTING: Radiology Department, University Hospital. SUBJECTS: Twenty four post-gastrectomy patients. INTERVENTION: A 22 cm phantom with a known amount of water and fat was scanned using high- and low-dose technique, standard and double table speed during a volumetric scan. The low-dose technique was implemented in the patient group. Total volume, total fat and four defined compartmental fat volumes in the truncal area were measured. RESULTS: The mean fat volume measured using the low-dose CT technique in the phantom was 0.2% above the actual fat content. The coefficient of variation for this method was 5%. By using low-dose, double speed instead of standard-dose technique, radiation exposure to the skin was decreased by more than 90% (equivalent to 4 mGy) of what is used in diagnostic imaging. The patient scans showed that no significant differences in BMI and total measured volume existed between female and male patients, but percent fat and percent subcutaneous fat were significantly larger in women (P = 0.006 and 0.002, respectively), as were percent intraabdominal and mediastinal fat in men (P = 0.002 and 0.003 respectively). CONCLUSIONS: Low-dose spiral CT accurately measures fat volume in vitro, and can be used in vivo for compartmental fat measurements. PMID- 9725662 TI - Breakfasts high in monoglyceride or triglyceride: no differential effect on appetite or energy intake. AB - OBJECTIVE: To test whether isoenergetic isoenergetically-dense doses of dietary 1 monoglyceride or triglyceride differentially influence appetite and meal-to-meal energy intake in man. DESIGN: Six men and six women were each studied twice in a 2 d protocol. On day 1 (maintenance day) they were fed a medium fat (MF) maintenance diet (MF: 40% fat, 47% carbohydrate and 13% protein by energy) calculated at 1.6 x resting metabolic rate (RMR). On day 2 (manipulation day) at 08.30 h subjects consumed a high-fat breakfast designed to contain 80-85% of RMR, composition 10% protein, 56% fat and 34% carbohydrate by energy, with 65% of energy for fat derived as either 1-monoglyceride or triglyceride. Food and energy intake were monitored at lunch (given at 12.30pm) and throughout the remainder of the day. During this time subjects had ad libitum access to isoenergetic, isoenergetically dense MF (40:47:13) foods (550kJ/100g), until 22.30pm). Subjective hunger and satiety were tracked hourly, during waking hours. RESULTS: There was no significant effect of fat type on food or energy intake at lunch or during the ad libitum period. There was no diet effect on subjective hunger (F(1,10)0.00; P= 0.975) in the inter-meal periods of morning or afternoon, nor during the whole day. Subjects found both diets to be similarly pleasant (F(1,61)0.84;P= 0.364). Men and women responded similarly, except that men ate more on all occasions than women. CONCLUSIONS: This study suggests that when a large dose of l-monoglyceride or triglyceride is incorporated into a breakfast meal, it behaves in a manner that is very similar to triglyceride in terms of the effects on hunger, appetite or feeding behaviour. PMID- 9725663 TI - Overfeeding fat as monoglyceride or triglyceride: effect on appetite, nutrient balance and the subsequent day's energy intake. AB - OBJECTIVE: To examine the effect of overfeeding isoenergetic diets enriched in 1 monoglyceride or triglyceride on nutrient oxidation and appetite throughout the day that it was given and the subsequent day's food and energy intake. DESIGN: Six men [mean (s.d.) weight; 76.89 (7.00) kg, height; 1.77 (0.05) m, age; 26.4 (6.0) y], were each studied twice in a 3 d protocol. On day 1 (maintenance day) they were fed a medium fat (MF) maintenance diet (MF: 40% fat, 47% carbohydrate and 13% protein by energy) calculated at 1.6 x resting metabolic rate (RMR). Subjects entered the calorimeter at 06.30 on day 2 for 49.5 h. On day 2 (manipulation day) subjects consumed a MF diet at 1.6 x RMR with an additional 0.45 x RMR as either 1-monoglyceride or triglyceride. On day 3 (outcome day), subjects had ad libitum access to isoenergetic, isoenergetically dense MF (40 :47: 13, 550 kJ/ 100 g) foods. Subjective hunger and satiety were tracked hourly, during waking hours throughout days 1-3. RESULTS: There was no significant effect of diet on nutrient oxidation or balance either during day 2 (manipulation day) or day 3 (outcome day), fat oxidation was similar on both diets. Subjective hunger was not affected by diet on either day with mean values of 34.3 and 35.0 mm (SED 5.2) on manipulation day (day 2) and outcome day (day 3), 35.3 and 40.8 mm (SED 5.2) on the 1 -monoglyceride or triglyceride diets respectively. Day 3 food and energy intake were unaffected by the previous day's dietary treatment, with mean intakes of 15.9 and 15.6 MJ (SED 1.07) on the 1-monoglyceride or triglyceride treatments, respectively. CONCLUSIONS: This study suggests that when 1-monoglyceride is covertly incorporated into a diet at unusually high levels, it behaves in a manner that is very similar to triglyceride, in its effects on appetite, feeding behaviour and net nutrient balance. PMID- 9725664 TI - Composition of body weight differences in subjects with the same body height. AB - OBJECTIVE: To determine the composition of body weight differences in men and women with the same height. DESIGN: A study of a random population sample of Danish subjects, aged 35-65 y. SUBJECTS: An age and sex stratified random sample of 3608 subjects of which 2987 (83%) subjects appeared for study. Of these, 548 men and 498 women were randomly selected for this present work based on their heights. RESULTS: The mean values of the slope of body fat against body fat free mass were 2.88 (s.e.m. = 0.09) in women and 1.84 (s.e.m. = 0.07) in men. There were no significant differences between the different body height classes. CONCLUSIONS: See results. PMID- 9725665 TI - Genetic heterogeneity of Indian field isolates of foot-and-mouth disease virus serotype O as revealed by partial sequencing of 1D gene. AB - The sequence of 165 nucleotides at the 3' end of the 1D gene, determined from RT PCR amplified cDNA fragments, of 25 type O strains isolated from different parts/regions of India during 1987 1995 and the vaccine strain (R2/75) currently in use in India were subjected to phylogenetic analysis. One isolate from the neighbouring country Nepal was also included in the study. The virus/ field strains showed high degree of genetic heterogeneity among themselves with % divergence in nucleotide sequence ranging from 1.2 to 19.4%. The Indian strains were much away (13.3 20.6%) from the exotic type O strains of O1BFS, O1K, and O1Campos. The type O strains analyzed were classified into three genotypes basing on level of divergence observed in nucleotide sequence. The type O vaccine virus (R2/75) was > 71% divergent (7.3-15.2%) from the field strains which revealed significant ( > 5%) genetic heterogeneity between the two. The phylogenetic analysis identified three distinct lineages, viz., (i) lineage 1 represented by the exotic strains, (ii) lineage 2 represented by 25 of the field strains which clustered into seven subgroups/sublines (2a-2g), and (iii) lineage 3 represented by a unique field isolate which shared the branching/origin with the vaccine strain. The lineage 2 which comprised of 25 of the 26 type O field strains analyzed, was placed almost at equidistance from the lineages 1 and 3 in the phylogenetic tree. The vaccine strain was closer to the viruses in lineage 2. Though there was no specific distribution pattern of sequences in different geographical regions of India, the viruses/ sequences in subgroup 2f appeared to be restricted to the southern states. Comparison of deduced amino acid sequence in the immunodominant regions 133-160 and 200-208 of the 1D gene product (VP1) showed that the two viruses in lineage 3 had unique amino acid residues at the positions 138 (D), 139 (G), 144 (I), and 158 (A) compared to rest of the strains including the exotic ones. Comparison of amino acid residues at critical positions 144, 148, 149, 151, 153, 154, and 208 revealed similarity between the type O strains analyzed. The virus strains showed variation (V/L/I) at position 144. One field strain showed replacement from Q149-->E and another from P208-->L. Thus, the study revealed that the type O FMD virus populations circulating in India and causing disease outbreaks are genetically much heterogeneous but related at the immunodominant region of VP1 polypeptide, and there are more than one genetically distinct virus populations in almost every region of the country which is possible due to unrestricted animal movement in the country. The involvement of vaccine virus in disease outbreaks was ruled out as the field strains (excluding the one in lineage 3) were phylogenetically distinct from it. PMID- 9725666 TI - Characterization of a recombinant human calicivirus capsid protein expressed in mammalian cells. AB - The capsid protein of the Hawaii strain of human calicivirus was expressed in the transient MVA/bacteriophage T7 polymerase hybrid expression system in order to examine its processing in mammalian cells. Selected amino acid modifications (an insertion, deletion, and substitution) at the predicted amino terminus of the capsid protein as well as the presence or absence of the ORF3 gene were examined for their effect on capsid expression. The protein was expressed efficiently in cell lines derived from three different species, with most of the expressed protein remaining localized within the cells. There was no evidence for N-linked glycosylation or myristylation of the 57 kDa capsid protein. Hawaii virus-like particles (HV VLPs), efficiently produced in the baculovirus expression system, were not observed in this expression system under the conditions in this study. PMID- 9725667 TI - Influence of host species on the evolution of the nonstructural (NS) gene of influenza A viruses. AB - The matrix (M) and nonstructural (NS) genes of influenza A viruses each encode two overlapping proteins. In the M gene, evolution of one protein affects that of the other. To determine whether or not this evolutionary influence operating between the two M proteins also occurs in the NS gene, we sequenced the NS genes of 36 influenza A viruses isolated from a broad spectrum of animal species (wild and domestic birds, horses, pigs, humans, and sea mammals) and analyzed them phylogenetically, together with other previously published sequences. These analyses enabled us to conclude the following host species-related points that are not found in the other influenza A virus genes and their gene products. (1) The evolution of the two overlapping proteins encoded by the NS gene are lineage dependent, unlike the M gene where evolutionary constraints on the Ml protein affect the evolution of the M2 protein (Ito et al.. J. Virol. 65 (1991) 5491 5498). (2) The gull-specific lineage contained nonH13 gull viruses and the non gull avian lineage contained H13 gull viruses, indicating that the gull-specific lineage does not link to the H13 HA subtype in the NS gene unlike findings with other genes. (3) The branching topology of the recent equine lineage (H7N7 viruses isolated after 1973 and H3N8) indicates recent introduction of the NS, M, and PB2 genes into horses from avian sources by genetic reassortment. PMID- 9725668 TI - Secretion and purification of HCV E1 protein forms as glutathione-S-transferase fusion in the baculovirus insect cell system. AB - We have expressed the E1 protein of Hepatitis C Virus (HCV) in a new recombinant form by using a baculovirus transfer vector directing the expression of proteins fused to the carboxy-terminus of glutathione-S-transferase (GST). The E1 domain was expressed varying at its carboxy terminus in order to retain (GST-E1) or delete (GST-E1b) the C-terminal hydrophobic region that may be involved in membrane association. Following infection with the recombinant virus, GST-E1b was efficiently secreted into the culture media and could be purified in a single step with the minimum of denaturation by glutathione affinity chromatography. The purified product was specifically immunoprecipitated by HCV positive human sera suggesting the maintenance of an immuno-relevant tertiary structure despite removal of the hydrophobic anchor. By contrast, cells infected with a recombinant baculovirus expressing GST-E1 gave a fusion protein with an appropriate molecular weight but also a series of polypeptides of lower molecular weight consistent with cleavage at the C-terminus of E1. GST-E1 was not secreted into the medium and was associated predominantly with the membrane fraction following cell disruption; the lower molecular weight forms were soluble and secreted. PMID- 9725669 TI - Characterization of the interaction of the human respiratory syncytial virus phosphoprotein and nucleocapsid protein using the two-hybrid system. AB - The interaction between the human respiratory syncytial virus phosphoprotein (P) and nucleocapsid (N) protein has been investigated using the two hybrid system in yeast and in tissue culture cells. Deletion analysis identified two regions in the P protein involved in this interaction. The immediate carboxy-terminal 20 amino acids were essential for interaction with the N protein. Point mutations in this region demonstrated that alteration of two conserved, phosphorylated, serine residues reduced binding to 50% of that of the native protein. The introduction of two proline residues to disrupt the predicted alpha-helical domain in this region dramatically reduced the ability of the mutant P protein to interact with the N protein. A second region which affected the interaction of the two proteins was located adjacent to the essential carboxy-terminal area. Deletion of this second region resulted in an increase in the strength of the interaction between the two proteins. These data shows that the RSV P protein, while having no amino acid sequence identity with the equivalent P protein of other negative strand viruses, is likely to have similar structural and functional features. PMID- 9725670 TI - Characterization of the structural proteins of hepatitis C virus expressed by an adenovirus recombinant. AB - Human adenoviruses have been used for mammalian expression vectors and recombinant vaccines for heterologous antigens. We constructed and characterized an infectious adenovirus recombinant containing core-E1-E2 genes of hepatitis C virus (HCV). The core protein was produced mainly during the early phase of viral infection. Expression of HCV E1 and E2 envelope proteins was detected by an immunoprecipitation with HCV-positive patient's sera. The purified E1 and E2 proteins appeared to be composed of mainly a heterodimeric form via noncovalent interaction, as previously observed in other mammalian expression systems. A small portion of E1 and E2 monomers as well as E1E2 aggregates by interdisulfide linkage were detected. Apparently heterodimeric E1E2 complexes were serologically reactive. The results suggest that adenovirus is an useful HCV antigen-expression vector. PMID- 9725672 TI - Identification of the 5' terminal structure of influenza virus genome RNA by a newly developed enzymatic method. AB - A combination of T4 polynucleotide kinase, Escherichia coli alkaline phosphatase, yeast Saccharomyces cerevisiae capping enzyme consisting of alpha (RNA guanylyltransferase) and beta (RNA 5'-triphosphatase) subunits. and its alpha subunit without RNA 5'-phosphatase activity was used to establish a simple enzymatic method for determination of RNA species with 5'-hydroxyl, 5' monophosphate, 5'-diphosphate or 5'-triphosphate termini. Using this method, we found that viral genome RNA (vRNA) segments of both A-type and C-type influenza viruses carry tri- or diphosphates at their 5' termini. The conclusion was based on the observations that: (i) 5' phosphorylation of vRNAs by T4 polynucleotide kinase takes place only after phosphatase treatment; and (ii) capping of vRNAs can be observed with both the intact yeast capping enzyme and its alpha subunit alone devoid of RNA 5'-triphosphatase activity; but (iii) the level of capping is higher for the alphabeta holoenzyme than the alpha subunit though the relative level varies depending on RNA preparations. The results support the de novo initiation for the RNA replication although transcription of influenza vRNAs is initiated by host cell capped RNAs as primers. PMID- 9725671 TI - A highly conserved genomic region in baculoviruses: sequence analysis of an 11.3 kbp DNA fragment (46.5-55.1 m.u.) of the Spodoptera exigua multicapsid nucleopolyhedrovirus. AB - A DNA fragment of 11.3 kilobase pairs (kbp) in size of the baculovirus Spodoptera exigua multicapsid nucleopolyhedrovirus (SeMNPV) genome (46.5 to 55.1 m.u.) was completely sequenced. Analysis of the sequence revealed eleven potential open reading frames (ORF). Ten of these ORFs showed significant amino acid identity to Autographa californica MNPV (AcMNPV) and Orgyia pseudotsugata MNPV (OpMNPV) genes p6.9, lef5, 38K, p19, p143, p25, p18, vp33, lef4, and vp39. One ORF (XC12) has no homolog in other baculoviruses and may be unique to SeMNPV. All but three of these putative genes are preceded by the consensus baculovirus late promoter element (5'-ATAAG-3'). The genetic organization and the putative map of transcripts of this fragment suggested that this region is highly similar to a region in AcMNPV fragment EcoRI-D. Comparison of the genetic organization of this 11.3 kbp fragment with the genomes of AcMNPV, OpMNPV, Bombyx mori NPV (BmNPV) and SeMNPV revealed that this region is highly conserved among baculovirus genomes. This is in contrast to the genetic organization of the polyhedrin-p10 region, which is much more diverged, but has been taken as point of reference to orient baculovirus physical maps. Through its diversity the latter region, however, would be an excellent candidate to determine baculovirus relatedness and phylogeny. The presence of conserved and diverged regions in baculovirus genomes with respect to gene order is reminiscent to the situation in other large DNA viruses, such as herpes- and poxviruses, where conserved central and diverged terminal parts are common characteristics. The role of this feature in the genomic organization of large DNA viruses is discussed with particular emphasis on virus replication and evolution. PMID- 9725673 TI - Infectivity of turnip mosaic potyvirus cDNA clones and transcripts on the systemic host Arabidopsis thaliana and local lesion hosts. AB - Full-length cDNA clones of turnip mosaic virus were assembled under the control of T7 or 35S promoter. The 35S or nopaline synthase terminator signals were introduced downstream the full length cDNA controlled by 35S promoter. Both the capped in vitro transcripts from T7 controlled template, and the cDNAs from 35S controlled plasmids were infectious on Arabidopis thaliana plants according to systemically induced symptoms and to ELISA assays. The cDNAs from 35S controlled plasmids induced local lesions in Chenopodium amaranticolor and Chenopodium quinoa plants. A spontaneous silent C/T transition, giving rise to an additional SpeI restriction site, not present in the original viral RNA template, was used as a marker of the origin of infection. PMID- 9725674 TI - Genomic heterogeneity maps to tandem repeat sequences in the herpes simplex virus type 2 UL region. AB - Two Bam HI Y and W fragments in the unique long sequence (UL) of the herpes simplex virus type 2 (HSV-2) were found to be heterogeneous in size among clones derived from a single strain as well as from epidemiologically unrelated isolates. More detailed restriction maps of these BamHI fragments were constructed and the heterogeneous subfragments were defined, cloned, and sequenced in order to investigate the mechanism causing the size difference. The subfragment of BamHI Y contained a tandem repeat sequence consisting of different numbers of 15 bp, 5'AGGGGCGGCTGGGGC3' as one unit among three isolates, and the subfragment of BamHI W contained the other tandem repeat sequence, 9 bp, 5'CCTCCCGCC3'. In the UL of the HSV-2 strain, these tandem repeat sequences were conserved and each repeat number appeared to be highly variable through viral genome replication. These results showed that the fragment length polymorphisms in these regions were attributable to the variation of unit numbers of the tandem repeat sequences. PMID- 9725675 TI - Traditional medicine and practices in burn care: need for newer scientific perspectives. PMID- 9725676 TI - Vancomycin resistant enterococci. PMID- 9725677 TI - Methicillin-resistant Staphylococcus aureus in burns patients--why all the fuss? AB - Procedures designed to limit spread of methicillin-resistant Staphylococcus aureus (MRSA) in burns units demand time and resources. To assess the significance of MRSA in burns patients we performed a retrospective review of MRSA colonization in in-patients over a 41-month period at the North Trent Sub regional Burns Unit. Patients were compared with MRSA free controls, matched for age and percentage body surface area (BSA) burn and admitted during the same time period. Length of stay, number of operations and deaths were outcome indicators. All patients managed non-operatively were excluded, leaving 40 MRSA patients and 46 controls. There was no statistical difference between the two groups with regard to number of operations (p= 0.07), duration of admission (p = 0.12) or mortality (p = 0.09). Of the control group, 83% had wound swabs positive for methicillin-sensitive Staphylococcus aureus (MSSA). there was no statistical difference in any outcome variables between this sub-group of controls and MRSA patients. Colonization with S. aureus (both MRSA and MSSA) was associated with larger burns (p<0.05), twice as many operative procedures (p<0.05) and prolonged admissions (p<0.01). Mortality was unaltered by staphylococcal colonization (p = 0.8). Although our study lacks power, we would suggest that methicillin resistance per se is not associated with increased morbidity or mortality in burns patients. PMID- 9725678 TI - Posttraumatic stress and maladjustment among adult burn survivors 1 to 2 years postburn. Part II: the interview data. AB - The purpose of this study was to assess the reaction to burn trauma, the hospitalisation characteristics and psychosocial adjustment in 174 burn victims who agreed to participate in a structured interview. It was postulated that once patients sustain a burn, the level of loneliness feelings and feelings of shame is a function of the visibility of the scars. Contrary to expectation, the visibility of the burn scars was not a predictor of pathological feelings of shame. By contrast, social introversion was a factor significantly associated with the development of burn related pathological feelings of shame. PMID- 9725679 TI - An investigation of the factors associated with an increased risk of psychological morbidity in burn injured patients. AB - Previous research aimed at identifying factors that increase the risk of major burns patients experiencing psychological problems post-burn has generally ignored the potential role of psychological factors. In a prospective study, patients with burn injuries ranging from < 1 per cent up to 40 per cent were interviewed within 2 weeks of sustaining the burn and followed up at ca 3 months post-burn in order to assess the effects of both non-psychological and psychological factors on their subsequent mental health. The factors investigated included burn related information, demographic information, previous psychiatric history, levels of psychological morbidity at 2 weeks post-burn, responsibility for the injury, previous life events, compensation claims and factors from the coping literature including appraisal, coping strategies and coping efficacy. Forward stepwise multiple regression analyses were used to investigate the relationships between these factors and subsequent mental health. Post-burn psychological morbidity was strongly associated with psychological factors including levels of psychological morbidity in the first 2 weeks of sustaining the injury and factors from the coping literature. PMID- 9725680 TI - Childhood burn injuries: circumstances of occurrences and their prevention in Ribeirao Preto, Brazil. AB - The objectives of this study were to verify which circumstances are present in burn accidents of patients under 12 years of age and to gather information to ground strategies to prevent those accidents. Parents or guardians of 26 patients aged under 12 years, admitted to the burns unit of the Clinical Hospital of Ribeirao Preto Medical School, Brazil, were interviewed, from March 1996 to March 1997. Fifty percent of the injured children were under 3 years of age and had suffered a scald. The kitchen and the backyard were identified as the places where the majority of accidents (84.6%) happened. At least one parent was present in 80.7% of cases. The results speak for the necessity of implementation of programs to prevent burn accidents, focusing on the domestic setting, chiefly activities in the kitchen. PMID- 9725681 TI - Scalds in children caused by water from electrical kettles: effect of prevention through information. AB - Electrical kettles (el-kettles) were virtually unknown in Danish households in the mid-1980s, but have since become more common. In 1996, (65 per cent of all Danish households had an el-kettle. As the number of el-kettles have increased, so have the number of scalds caused by water from toppled el-kettles. The first patient with an el-kettle scald was admitted to the Burns Centre at Hvidovre hospital in 1988. From 1988 to 1993 29 patients were admitted with this type of scald; 15 patients in 1993 alone. All the patients were toddlers 5-30 months of age. When el-kettle scalds were compared to scalds caused by other mechanisms in children under 5 years of age, it was found that the former occurred to younger children than the latter (60 per cent of el-kettle scalded children were less than 1 year of age compared to 28 per cent scalded by other means), the scalds were more extensive (median of TBSA were 13 per cent and 6 per cent, respectively), and the scalds were deeper (53 per cent and 24 per cent, respectively, needed skin-grafting). In 1993 campaigns were started to inform parents that the cord of the el-kettle should be short and not hang over the edge of the table. In the following years a considerable decrease in the number of el kettle scalds was found. When the number of expected el-kettle scalds was estimated from the number of Danish households having an el-kettle, it was found that more than half the expected number of el-kettle scalds were avoided. PMID- 9725682 TI - Relationship between parental emotional states, family environment and the behavioural adjustment of pediatric burn survivors. AB - The purpose of this study was to examine the relationship between psychosocial adjustment of the burned child and characteristics of the child's family. It hypothesized that parents who perceived their children without major behavioural problems would possess supportive family values and would, themselves, be better adjusted psychologically than those parents who perceived their children as possessing multiple behavioural problems. A stratified random sampling technique was used to select 35 (29 boys, 6 girls) paediatric burn survivors, ages 9 to 18, 1-5 years post-burn, with burn sizes ranging from 3 to 92% burn. Subjects' parents were administered the Child Behaviour Checklist (CBCL), the Family Environment Scale (FES), the Impact of Events Scale (IES), and the Beck Depression Inventory (BDI). The subjects were divided into two groups on the basis of the total problem CBCL scores. i.e. troubled (T > or = 60) or untroubled (T<60). One-way ANOVA tests revealed no significant differences between the two groups in the way parents reacted to trauma (IES) and parental depression (BDI). Significant differences (p<0.01) were revealed between the two groups on FES subscales. The parents of the untroubled group scored higher on 'Cohesion' and 'Organization' and lower on 'Conflict'. These parents also scored higher (p< or =0.05) on 'Achievement Orientation'. The results indicate that work with the family to promote cohesion. to decrease conflict, to enhance stability and to promote expectation of positive achievement must he a part of the rehabilitation of the burned child. PMID- 9725683 TI - Epidemiological data and mortality rate of patients hospitalized with burns in Brazil. AB - This retrospective analysis of burn patients in a University Hospital in the state of Sao Paulo, Brazil, was carried out to characterize this population and to identify the factors that affect the mortality rate. All patients hospitalized from January 1990 to April 1995 (n = 229, 3.6 patients/month) and who terminated treatment were included. Of these, 80.8% (185 patients) were hospitalized within 24 h of the burn. Occupational and/or domestic accidents were responsible for most of the burns (78.6%), which were mainly caused by a direct flame (71.2%). with alcohol being the flammable fluid most frequently used. The average patient treated at the center was a male of 9 years of age or less with 20-40% burned body surface, who received care within 24 h after suffering an accidental alcohol burn and who was hospitalized for < or =30 days. The mortality rate was 18.8% for all patients and increased with burned body surface and age, and for suicide patients. Suicide attempts for all patients > or = 18 years were the cause of 46 .5% (20/43) of the burns involving women and of 8.9% (8/90) of the burns involving men. The mortality rate was significantly higher for self-inflicted burns (42.9%) than for accidental burns (20.2%). PMID- 9725684 TI - Preventable burns associated with the misuse of gasoline. AB - Gasoline is intended for use as a motor fuel, but the universal availability of gasoline in the home encourages misuse as a solvent, insecticide, accelerant or cleaning solution. The careless or inappropriate use of gasoline may result in burn injury. We examined the circumstance of gasoline-related injury in a population admitted to one burn centre to determine the potential for burn prevention efforts. A retrospective review of all burn admissions to one centre for the years 1978 to 1996 demonstrated hat 1011 of 4339 acute admissions (23.3%) were gasoline-related. This group had an average total burn size of 29.8% total body surface (TBSA) and an average full thickness injury of 14.4% TBSA. There were 144 fatalities resulting from gasoline-associated burn injury. Where such determination could be made, the use of gasoline was judged to be inappropriate or unsafe in 687 of 788 cases (87.1%). Ninety of 144 fatalities (62.5%) were associated with inappropriate or unsafe use of gasoline. The careless or inappropriate use of gasoline poses significant risk of burn injury. The indoor use of gasoline, as well as use of gasoline for purposes other than as a motor fuel, should be strongly discouraged. PMID- 9725685 TI - Industrial burns in Jamshedpur, India: epidemiology, prevention and first aid. AB - Industrial burn injuries result in significant morbidity, infrequent mortality and man-hour loss, leading to loss of productivity. With a view to study the epidemiology of industrial burns in Jamshedpur and first aid awareness, we analysed 815 patients (142 inpatients and 673 outpatients) with industrial burns seen by us during the period from January 1993 to December 1996. 69% of these injuries were caused by contact with hot objects, while the rest were caused by flame, electrical and chemical agents. Our audio-visual awareness promotion programme for burns safety and first aid awareness amongst our officers and employees at various levels, has been successful in reducing the incidence of burns. The campaign has also popularised the use of cool water as the best first aid measure. PMID- 9725686 TI - Background pain in burn patients: routine measurement and recording of pain intensity in a burn unit. AB - It goes without saying that pain following a burn must be treated but it is not so evident to measure and document the intensity of pain and the efficacy of treatment. Since 1994 the authors have routinely measured background pain, that is, at rest, along with temperature and pulse rate. For analysis and quality assessment a relational database programme is used in the ward. In this paper the authors' experience is reported from a consecutive series of 98 patients with burn injuries who assessed the intensity of pain on a visual analogue scale. There were great intra- and inter-individual variations in pain intensity. Highest values were found during the first week of treatment when female patients experienced pain more intensively than male. For other time periods there was no statistical significant difference between the sexes. Pain intensity and severity of burn was not related except during the second week when patients with major burns had a tendency to express more pain than moderate burns. Measurement of background pain along with other routine registrations is easy and not time consuming. Patients needing intensified pain treatment can be identified. For research and quality assessment a computerized patient register is of great help. PMID- 9725687 TI - Nitric oxide synthesis in myocardium following burn injury in rats. AB - We investigated nitric oxide and cyclic GMP production in myocardium early after burn injury in rats. Nitric oxide synthase activity was measured in cytosol from the left ventricular wall of burned rats. Cytosol from control group animals was shown to contain mainly Ca2+-dependent nitric oxide synthase (cNOS) with a small amount of Ca2+-independent nitric oxide synthase (iNOS). Following burn injury, there was a marked increase in iNOS activity with a peak at 8 h post-burn, however, myocardial cNOS activity was found to decline. Parallel to iNOS induction there was a significant increase in myocardial nitric oxide and cyclic GMP production. All these changes were alleviated by treatment of the rats with dexamethasone. Since increases in cyclic GMP levels in the heart were associated with reduced myocardial contractility, it is possible that enhanced production of nitric oxide by a Ca2+-independent NO synthase accounts, at least in part, for the depression of myocardial contractility seen in burn animals and patients. PMID- 9725688 TI - Topical D-myo-inositol-1,2,6-trisphosphate and hexylbetaine treatment reduces albumin extravasation in experimental rat skin burn injury. AB - The anti-inflammatory agent D-myo-inositol-1,2,6-trisphosphate (1,2,6-IP3) has shown beneficial effects in experimental burns following systemic administration. The purpose of this study was to investigate the effect of topical 1,2,6-IP3 cream on a standardised full-thickness 1 cm2 burn injury in rats. The experimental cream contained a transcutaneous absorption enhancer, hexylbetaine. Five different treatment groups were used. Two experimental groups of burned rats received either 1,2,6-IP3 cream with hexylbetaine (n = 10) or without hexylbetaine (n = 10). Two burned control groups were treated either with hexylbetaine cream (n = 10) or placebo cream (n = 10), while a third control group was untreated (n = 14). The various creams (0.5 g) were administered to the experimental burn area and allowed to remain for 3 h covered with an occlusive dressing. Spectrophotometrical quantification of Evans blue albumin extravasation was used to evaluate the effect of the experimental creams on vascular permeability following the burn trauma. Results showed a significant reduction of albumin extravasation both by 1,2,6-IP3 (p<0.05) and by hexylbetaine alone (p<0.01), as compared to placebo cream-treated animals. The transcutaneous absorption enhancer hexylbetaine did not further improve the effect of 1,2,6-IP3 on burn oedema. In conclusion, both topical 1,2,6-IP3 and hexylbetaine induced a significant reduction of albumin extravasation in burned skin. The effect of 1,2,6-IP3 could be related to previously shown anti-inflammatory actions of the agent, while the mechanisms of actions of hexylbetaine remain to be investigated. PMID- 9725689 TI - Apoptosis of hair follicle cells in the second-degree burn wound unders hypernatremic conditions. AB - Progressive burn wound necrosis is an important factor as a cause of delayed healing during clinical therapy of burns. Among the causes of progressive necrosis have been attributed an insufficient blood supply or a dehydration at the zone of stasis just beneath the zone of coagulation. In a previous study evidence was presented that hypernatremia, an osmotic injury, may act to promote progressive tissue or cell death of the superficial dermal wound resulting from a heat injury. To test this hypothesis pathological features of cell death in the second-degree burn wound in the rat with hypernatremia were investigated and evidence for apoptosis in hair follicle cells was observed. Rats in the hypernatremic group were administered 10 ml of hypertonic sodium solution (850 meq 1(-1)) and the control rats were treated with 10 ml of hyponatremic solution (100 meq 1(-1)) to prevent hypernatremia. After 24 h postburn the average incidence of hair follicles (ratio to the normal skin) in the hypernatremic group was 30.1 +/-11.6 per cent and significantly lower when compared with the control group (87.6+/-6.0 per cent). The numbers of hair follicles were studied by haematoxylin and eosin stain, and the apoptotic process was investigated by an immunochemical assay and electron microscopy. PMID- 9725690 TI - Oral fluid therapy in paediatric burns (5-10%): an appraisal. AB - Fluid therapy by the oral route is the accepted method of treatment for smaller burns in children (less than 10%). [Settle JAD. Burns - the first five days. Essex: Smith and Nephew Pharmaceuticals Ltd, 1986.] A phone survey was carried out of all the hospitals in the United Kingdom that manage burns, to record their oral fluid therapy practices for burns (5-10% BSA) in paediatric patients. Included in the survey was an assessment of the uniformity of the contents of the fluids, their palatability and acceptance by patients and any side-effects from this form of treatment. There appears to be no uniformity in policies regarding fluid therapy in children with this percentage of burns. Treatment ranged from a formula guided resuscitation therapy (as practised generally with large burns) to a 'drink as you like' policy. Fluids used varied from electrolyte to non electrolyte containing solutions and fruit juices and were, therefore, markedly different in content. The electrolyte solutions were reported as being non palatable unless flavoured with fruit juices. No complication was reported although one unit queried a possible case of fluid overload. Potential complications from this mode of therapy are discussed. The cost implications of using various fluid types are also presented. PMID- 9725691 TI - Burn wound excision and massive blood transfusion did not affect perioperative vancomycin levels. AB - INTRODUCTION: The effect of burns surgery that requires intraoperative transfusion of five or more units of blood on serum vancomycin levels was assessed. METHODS: Serum vancomycin levels were measured 10 min and 6 h after vancomycin administration with surgery being performed in the interval. The following day the same dose of vancomycin was given and serum vancomycin levels measured at the same times without intervening surgery. RESULTS: Thirteen operations involving nine patients who required a mean blood transfusion of 9.2 units were studied. There was very little difference between 10-min levels, 6-h levels and the change over interval (absolute and percentage) on the day of surgery and the following day. The recorded serum levels were often at the lower end of the desired range, especially in patients who underwent the larger operations. This was the case on both day of surgery and the control day. CONCLUSIONS: Large volume blood loss and replacement during burns surgery did not significantly affect perioperative vancomycin levels. 1998 Elsevier Science Ltd for ISBI. PMID- 9725692 TI - A burn-like lesion caused by a testosterone transdermal system. AB - A wide range of drugs are capable of being administered as transdermal drug delivery systems (TDDS). Despite their advantages, these systems are known to have adverse effects. This report describes localised skin reactions with reference to a burn-like lesion on the shoulder of a patient caused by a testosterone TDDS. Patients should be advised of the potential adverse effects of these systems and also of suitable application sites, avoiding areas liable to prolonged pressure. PMID- 9725693 TI - Citric acid treatment of severe electric burns complicated by multiple antibiotic resistant Pseudomonas aeruginosa. AB - A case of severe electric burns complicated by multiple antibiotic resistant Pseudomonas aeruginosa not responding to various antibiotics administered systemically is presented. Citric acid (3%) was used successfully to eliminate Pseudomonas aeruginosa from burn wounds and infection was completely controlled in 14 days. Citric acid treatment is evidently of value in the clinical control of burn wound colonization caused by difficult strains of Pseudomonas aeruginosa. PMID- 9725694 TI - Burn management in a patient with autism. AB - The successful use of elective post-operative sedation and ventilation following tangential surgical excision of burns in an autistic child. non-compliant with conventional management is reported. Other strategies for treating autistic children are discussed. PMID- 9725695 TI - Hypopigmentation along subcutaneous veins following intrakeloid triamcinolone injection: a case report and review of literature. PMID- 9725696 TI - Recent references. PMID- 9725697 TI - Expression of L1 and PSA during sprouting and regeneration in the adult hippocampal formation. AB - The objective of the present study was to evaluate the expression of polysialic acid (PSA) and the cell adhesion molecule L1 during axonal regeneration and sprouting after injury to the adult rat brain. All animals received a complete lesion of the fimbria-fornix (FF). Grafts of nerve growth factor (NGF)- or beta galactosidase (betaGal)-producing fibroblasts were placed in the FF lesion cavity and induced septohippocampal cholinergic regeneration or sympathetic tyrosine hydroxylase (TH)-positive sprouting, respectively. Cholinergic regeneration was evaluated from 2 to 8 weeks following grafting of NGF-producing fibroblasts in the FF lesion cavity. In the graft area, choline acetyltransferase (ChAT) positive fibers expressed L1 and PSA. Once cholinergic axons reached the hippocampal formation (HF), they no longer expressed L1 or PSA. Eight weeks after a lesion of the FF and transplantation of betaGal-producing fibroblasts, TH positive fibers sprouted in the denervated HF and expressed L1 but not PSA. At the zone of reactive gliosis, PSA but not L1 expression was increased following a lesion of the FF and transplantation of NGF- or betaGal-producing fibroblasts. In animals that received a graft of NGF-producing fibroblasts in the FF lesion cavity, numerous ChAT-positive axons were observed along these areas rich in PSA and reactive astrocytes. Taken together, these results suggest that the expression of PSA and L1 is upregulated on regenerating cholinergic axons during axonal elongation and downregulated upon target innervation. In contrast, TH positive fibers that sprout in the denervated HF express and maintain their expression of L1. Finally, the expression of PSA in the area of reactive gliosis may contribute to a permissive environment for axonal regrowth. PMID- 9725698 TI - Phenotypic conversion of distinct muscle fiber populations to electrocytes in a weakly electric fish. AB - In most groups of electric fish, the electric organ (EO) derives from striated muscle cells that suppress many muscle phenotypic properties. This phenotypic conversion is recapitulated during regeneration of the tail in the weakly electric fish Sternopygus macrurus. Mature electrocytes, the cells of the electric organ, are considerably larger than the muscle fibers from which they derive, and it is not known whether this is a result of muscle fiber hypertrophy and/or fiber fusion. In this study, electron micrographs revealed fusion of differentiated muscle fibers during the formation of electrocytes. There was no evidence of hypertrophy of muscle fibers during their phenotypic conversion. Furthermore, although fish possess distinct muscle phenotypes, the extent to which each fiber population contributes to the formation of the EO has not been determined. By using myosin ATPase histochemistry and anti-myosin heavy chain (MHC) monoclonal antibodies (mAbs), different fiber types were identified in fascicles of muscle in the adult tail. Mature electrocytes were not stained by the ATPase reaction, nor were they labeled by any of the anti-MHC mAbs. In contrast, mature muscle fibers exhibited four staining patterns. The four fiber types were spatially arranged in distinct compartments with little intermixing; peripherally were two populations of type I fibers with small cross-sectional areas, whereas more centrally were two populations of type II fibers with larger cross-sectional areas. In 2- and 3-week regenerating blastema, three fiber types were clearly discerned. Most (> 95%) early-forming electrocytes had an MHC phenotype similar to that of type II fibers. In contrast, fusion among type I fibers was rare. Together, ultrastructural and immunohistochemical analyses revealed that the fusion of muscle fibers gives rise to electrocytes and that this fusion occurs primarily among the population of type II fibers in regenerating blastema. PMID- 9725699 TI - Absence of plasticity of the frequency map in dorsal cochlear nucleus of adult cats after unilateral partial cochlear lesions. AB - In adult animals, lesions to parts of the auditory receptor organ, the cochlea, can produce plasticity of the topographic (cochleotopic) frequency map in primary auditory cortex and a restricted or patchy plasticity in the auditory midbrain. This effect is similar to the plasticity of topographic maps of the sensory surface seen in visual and somatosensory cortices after restricted damage to the appropriate receptor surface in these sensory systems. There is dispute about the extent to which subcortical effects contribute to cortical plasticity. Here, we have examined whether topographic map plasticity similar to that seen in the auditory cortex and the midbrain is observed in the adult auditory brainstem. When partial cochlear lesions were produced in the same manner as those that were produced in the cortex and midbrain studies, we found no plasticity of the frequency map in the dorsal cochlear nucleus (DCN). Small regions of the DCN that were deprived of their normal, most sensitive frequency (characteristic frequency; CF) input by the cochlear lesion appeared to have acquired new CFs at frequencies at or near the edge of the cochlear lesion. However, examination of thresholds at the new CFs established that the changes simply reflected the residue of prelesion input to those sites: The patterns of CF thresholds were very well predicted by simple calculations of the patterns that were expected from such residual input. The results of this study suggest that the DCN does not exhibit the type of plasticity that has been found in the auditory cortex and midbrain; therefore, it does not account for the changes in responsiveness observed in the higher level structures under similar experimental conditions. PMID- 9725700 TI - Morphological development of afferent segregation and onset of synaptic transmission in the trigeminothalamic pathway of the wallaby (Macropus eugenii). AB - A light and electron microscopic study has been made of the time of formation of whisker-related patterns in trigeminothalamic afferents and the onset of synapse formation between afferents and cells in the ventroposteromedial nucleus (VPM) of the marsupial mammal, the wallaby, by labelling afferents with a carbocyanine dye. A parallel in vitro study was made of the functional development of the trigeminothalamic pathway to the VPM. Evoked synaptic responses could be recorded in the VPM from the time that afferents first reached the VPM at postnatal day 15 (P15). At all stages, the excitatory response comprised both N-methyl-D-aspartate and non-N-methyl-D-aspartate-mediated components. At P40, the response decreased markedly in duration, coinciding with the onset of synaptogenesis. This implies that transmission is occurring prior to synapse formation, probably through transmitter release from growth cones. At P50, synaptic responses became dominated by a fast, non-N-methyl-D-aspartate potential, and this coincided with the first appearance of whisker-related patterns in the VPM. A gamma-aminobutyric acid (subtype A)-mediated, inhibitory component was also present from the time of afferent arrival. These findings support the idea that functional interactions between afferents and their targets may play a role in pattern formation in the somatosensory thalamus. PMID- 9725701 TI - Projection patterns from the raphe nuclear complex to the ependymal wall of the ventricular system in the rat. AB - The goal of the present study was to characterize the anatomical and neurochemical relationships that the raphe nuclear complex maintains with respect to lateralized and centralized components of the ventricular system. From this investigation, we 1) determined the ipsilateral vs. contralateral distribution of raphe efferents to the ependymal wall of the lateral ventricle, 2) assessed the degree of collateralization of individual ependymal projection neurons to other sites along the ventricular path, 3) compared the topography of raphe neurons that project to the ventricular lining as well as the lumen of the fourth and lateral ventricles, and 4) evaluated the neurochemical identity of raphe neurons that innervate the ventricular system. After tracer injections into the lateral ventricle, labeled cells were distributed evenly on both sides of the midline in the dorsomedial subregion of the intermediate dorsal raphe nucleus. Further rostrally, labeled cells were clustered bilaterally above the medial longitudinal fasciculi and extended into the median raphe nucleus. Injections that involved the ependymal wall of the lateral ventricle resulted in prominent ipsilateral labeling within the dorsal raphe nucleus, just ventral to the cerebral aqueduct. Most of the labeled cells in this latter group had collateral projections to other sites along the ventricular path. Most of the ventricle projection cells contained serotonin but not nicotinamide adenine dinucleotide phosphate diaphorase. These findings indicate that the raphe nuclear complex is topographically organized with respect to the ventricular system. Selected subsets of serotoninergic dorsal raphe neurons may influence discrete segments of the ventricular system independently as well as coordinate functions throughout the system through axon collaterals to other sites along the ventricular neuraxis. PMID- 9725702 TI - Neuronal projections from the mesencephalic raphe nuclear complex to the suprachiasmatic nucleus and the deep pineal gland of the golden hamster (Mesocricetus auratus). AB - Neuronal projections from the mesencephalic raphe system to the suprachiasmatic nucleus and the pineal complex were mapped in this study of the golden hamster, by use of the anterograde tracer Phaseolus vulgaris-leucoagglutinin and the retrograde tracer cholera toxin subunit B. From the median raphe nucleus, a rostral projection ascended in the ventral part of the mesencephalon to continue in the medial forebrain bundle of the forebrain. Nerve fibres from this bundle innervated the ventral and medial parts of the suprachiasmatic nucleus. At the level of the interpeduncular nucleus of the mesencephalon, fibres of the ventral bundle bent dorsally to reach the epithalamic area and to continue in the forebrain in a periventricular position. Some of these fibres innervated the dorsal tip of the suprachiasmatic nucleus. The dorsal raphe nucleus was the origin of a nerve fibre bundle, located in the periaqueductal gray of the mesencephalon, innervating the deep pineal gland and pineal stalk. Injection of cholera toxin B into the suprachiasmatic nucleus labelled cells in the median raphe. Combination of the retrograde tracing from the suprachiasmatic nucleus and serotonin transmitter immunohistochemistry showed that some of the cholera toxin B-immunoreactive nerve cells also contained serotonin. Thus, this study of the golden hamster shows a serotonergic projection from the median raphe nucleus to the suprachiasmatic nucleus and a projection from the dorsal raphe nucleus to the deep pineal gland supporting physiological indications of an influence of serotonin on the photoreceptive circadian system of the brain. PMID- 9725703 TI - Oligodendrocytes are not inherently programmed to myelinate a specific size of axon. AB - Current studies support the morphological classification of oligodendrocytes proposed by Del Rio Hortega ([1922] Bol. R. Soc. Esp. Hist. Nat. 10:25-29; [1924] C.R. Soc. Biol. 91:818-820), in which cells either myelinate multiple internodes that are associated with small axons, or they myelinate restricted/single internodes of large-diameter axons. The reasons why an oligodendrocyte myelinates a particular calibre of axon are unknown. Because progenitors are generated in restricted, subventricular zones, an intrinsic program would imply that germinal centres contain a mixture of cells, each committed to myelinate axons of a particular size. Conversely, each cell could have the potential ability to myelinate any size axon. We tested this latter hypothesis that oligodendrocyte progenitors are uncommitted in their ability to myelinate a particular axon size. We introduced oligodendrocyte lineage cells from the optic nerve, which normally encounter only small-diameter axons, to a myelin-deficient environment containing a large range of axon sizes. Dissociated, mixed glial cells from the optic nerve were characterised immunocytochemically and were grafted into the spinal cord ventral column of neonatal, myelin-deficient rat mutants. Examination of the patches of myelin produced by these cells at different times after transplantation revealed that optic nerve oligodendrocytes were capable of producing a widespread, nonselective myelination of axons that were destined to have both small or large calibres. Thus, an axonal or local signal, and not an intrinsic program, is probably responsible for the previously described oligodendrocyte diversity. PMID- 9725704 TI - Oxytocinergic inputs to the nucleus of the solitary tract and dorsal motor nucleus of the vagus in neonatal rats. AB - The paraventricular nucleus of the hypothalamus (PVN) modulates vagal digestive motor functions via oxytocinergic projections to the nucleus of the solitary tract (NST) and dorsal motor nucleus of the vagus (DMV) in adult rats. Little is known regarding the structural or functional maturation of these projections. The present study examines the postnatal development of immunocytochemically identified oxytocinergic fibers in gastric subregions of the medial NST-DMV. For this purpose, a monoclonal antibody (PS36) that recognizes both oxytocin (OT) neurophysin and its prohormone was used to identify oxytocinergic fibers. PS36 positive fibers already were present within the NST-DMV in rats on the day of birth. Retrograde transport of cholera toxin neural tracer from the NST-DMV in newborn rats confirmed that PVN neurons were the sole source of these oxytocinergic fibers. The cumulative length of PS36-positive fibers in sampled subregions of the medial NST and DMV increased approximately 23-fold and 94-fold, respectively, between birth and adulthood. The observed postnatal increases in PS36 immunolabeling could reflect increased delivery of immunoreactive antigen from hypothalamic perikarya to distal axons and/or increasing oxytocinergic innervation of the NST-DMV. Additional work will be needed to address these questions and to determine the time course during which central oxytocinergic pathways become mature in their ability to influence vagally mediated digestive functions. PMID- 9725705 TI - Postnatal cytoarchitecture of the rat medial geniculate body. AB - The medial geniculate body (MGB) is a thalamic structure that provides vital information flow to the forebrain for complex acoustic processing. The development of cytoarchitectural features of the MGB was examined in rat to identify age-related patterns of growth in major geniculate compartments that have been described previously (Clerici and Coleman [1990] J. Comp. Neurol. 297:14-31; Clerici et al. [1990] J. Comp. Neurol. 297:32-54): the ventral (MGv), dorsal (MGd), and medial (MGm) divisions. Results show that, on the day of parturition, all major nuclei of each division are characterized, including the ovoid (OV) and ventral (LV) nuclei of MGv; the dorsal, deep dorsal (DD), caudodorsal, limitans, and suprageniculate nuclei of MGd; and the MGm. The MGv and MGd, which display comparable areas at birth, show rapid growth to postnatal day 7 (PND7), which then slows until PND11, around the time of ear canal opening; subsequently, MGv accelerates growth to reach larger adult size. From PND11 to PND16, thionin facilitates parcellation by extensive staining of dendritic processes of MGd, MGm, and lateral posterior nucleus neurons but not neurons of the MGv or the dorsal lateral geniculate nucleus. Golgi stains after birth reveal restricted dendritic arborizations in MGv cells and dichotomous branching patterns of MGd neurons. Somal size in MGB increases dramatically subsequent to afferent innervation and again following onset of auditory function. Somal growth occurs between all postnatal age groups tested for OV, LV, and DD nuclei, although LV segments related to high and low frequencies do not differ. Cell packing density predicts the expanse of major MGB divisions better than somal size. These results demonstrate the integrity and growth patterns of MGB nuclei and divisions from nascence and provide a substrate for subsequent study of anatomical and physiological development of the MGB. PMID- 9725706 TI - Reorganization of sensory modalities evoked by microstimulation in region of the thalamic principal sensory nucleus in patients with pain due to nervous system injury. AB - Stimulation of the somatosensory system is more likely to evoke pain in patients with chronic pain after nervous system injury than in patients without somatosensory abnormalities. We now describe results of stimulation through a microelectrode at microampere thresholds (threshold microstimulation; TMIS) in the region of the human thalamic principal sensory nucleus (ventral caudal; Vc) during operations for treatment of movement disorders or of chronic pain. Patients were trained preoperatively to use a standard questionnaire to describe the location (projected field) and quality of sensations evoked by TMIS intraoperatively. The region of Vc was divided on the basis of projected fields into areas representing the part of the body where the patients experienced chronic pain (pain affected) or did not experience chronic pain (pain unaffected) and into a control area located in the thalamus of patients with movement disorders and no experience of chronic pain. The region of the Vc was also divided into a core region and a posterior-inferior region. The core was defined as the region above a standard radiologic horizontal line (anterior commissure posterior commissure line; ACPC line) where the majority of cells responded to innocuous somatosensory stimulation. The posterior-inferior area was a cellular area posterior and inferior to the core. In both the core and the posterior inferior regions, the proportion of sites where TMIS evoked pain was larger in pain-affected and unaffected areas than in control areas. The number of sites where thermal (warm or cold) sensations were evoked was correspondingly smaller, so that the total of pain-plus-thermal (sensation of warmth or cold) sites was the same in all areas. Therefore, sites pain where stimulation evoked pain in patients with neuropathic pain (i.e., pain following an injury to the nervous system) may correspond to sites where thermal sensations were evoked by stimulation in patients without somatosensory abnormality. PMID- 9725707 TI - Distribution of prepro-nociceptin/orphanin FQ mRNA and its receptor mRNA in developing and adult mouse central nervous systems. AB - Nociceptin/orphanin FQ (N/OFQ) and its receptor share similarities to opioids and their receptors in terms of the molecular structure and signaling pathway, but the two systems exhibit different actions in vivo. To understand the mechanism of N/OFQ-system actions, we examined, by in situ hybridization analysis, the distribution of preproN/OFQ and N/OFQ receptor mRNAs in the developing and adult mouse central nervous systems (CNS). In most neural regions, preproN/OFQ mRNA was mainly expressed in a small population of middle-sized neurons. These neurons were scattered between large projection-type neurons or within the neuropil, suggestive of interneurons. In some other nuclei (lateral septum, bed nucleus of the stria terminalis, reticular thalamic nucleus, inferior colliculus, and rostral periolivery nucleus), preproN/OFQ mRNA was expressed in a number of large projection-type neurons. By contrast, N/OFQ receptor mRNA was evenly expressed in most neurons of the adult CNS. Considering the inhibitory actions of N/OFQ, the distinct cellular expression pattern of the N/OFQ system suggests that the release of N/OFQ from interneurons may lower neuronal and synaptic activities of neighboring neurons, leading to integration or modulation of local circuits. Furthermore, the cellular expression pattern, distinct from that of the opioid system, may provide a possible molecular/cellular basis for the different in vivo actions of N/OFQ and opioids. In embryonic stages, both preproN/OFQ and N/OFQ receptor mRNAs were highly and widely expressed in the mantle zone, suggesting the possible importance of N/OFQ signaling in CNS development. PMID- 9725708 TI - Pituitary expression of protein kinase C isotypes during early development. AB - Protein kinase C (PKC) is a critical regulator of signal transduction and cell function in many tissues, including pituitary. Although PKC influences pituitary hormone secretion in adults, its role in determining characteristic perinatal patterns of hormone secretion and synthesis is not known, and the expression of major PKC isotypes in perinatal pituitary is poorly defined. We therefore determined the developmental, cell-specific expression of the major PKC isotypes, using Western analysis and double label immunohistochemistry, in pituitaries of perinatal and mature rats. Expression of specific PKC isotypes was strikingly age dependent. Pituitary expression of PKC alpha was particularly high in neonates and declined significantly with age, with levels in adult rats approximately half those of neonates as assessed by Western analysis. Similarly, immunohistochemistry indicated that PKC alpha was less abundant in adult than in neonatal pituitaries; the most intensely staining cells of both age groups were identified as somatotrophs and gonadotrophs. In contrast to PKC alpha, pituitary expression of PKC epsilon increased approximately two-fold with advancing age as assessed by Western analysis; this age-dependent pattern was confirmed by immunohistochemistry. Perinatal pituitaries expressed PKC epsilon in some somatotrophs and in all gonadotrophs, whereas PKC epsilon expression was limited to gonadotrophs in the mature pituitary. Pituitary expression of PKC betaII, delta, and zeta did not differ with age, and PKC gamma was not detected in pituitaries of any age group. These results indicate that expression of PKC isotypes within the pituitary is developmentally regulated in a cell-specific and isotype-specific manner, and are consistent with the concept that PKC contributes to the regulation of pituitary function during early development. PMID- 9725709 TI - Intracranial androgenic activation of male-typical behaviours in house mice: concurrent stimulation of the medial preoptic area and medial nucleus of the amygdala. AB - This experiment examined whether testosterone proprionate (T) action in the medial preoptic area (MPO) would synergize with T action in the medial nucleus of the amygdala (AME) for the expression of androgen-dependent behaviors in house mice. Cannulae containing T were bilaterally implanted into the MPO, the AME, or both areas concurrently (MPO/AME) of castrated males. In addition, other castrates were implanted subcutaneously with empty Silastic capsules (BSIL) or Silastic capsules containing T (TSIL). All subjects were examined for the following androgen-dependent, male-typical behaviors: mounting, urinary scent marking, preference for female urine over male urine, preference for female over male conspecifics and ultrasonic mating vocalizations. MPO implants restored ultrasonic vocalizations and preference for females, but had little or no effect upon urine marking, mounting or preference for female urine. In contrast, AME implants were ineffective at restoring any of these male-typical behaviors. The combined MPO/AME implants were not more effective in restoring male-typical behaviors than MPO implants alone, thus providing no evidence for synergy in hormone action between these two brain areas. In general, castration (BSIL) resulted in low levels of all behaviors whereas systemic T replacement (TSIL) resulted in high levels of behavior, verifying the androgen-dependence of these behaviors. Group differences in male-typical behavior could not be accounted for by differences in general activity levels. Moreover, none of the brain-implanted groups had larger seminal vesicles than those of the BSIL. Thus, when the brain implants affected behavior, they most probably did so through their effects within the brain. Although the AME is a target for steroid hormones and is an important area for the expression of male-typical behaviors, intracranial T implants into the AME did not demonstrate a role for androgen in the AME in restoring male-typical behaviors in castrated mice. PMID- 9725710 TI - Photoperiod affects the ability of testosterone to alter proopiomelanocortin mRNA, but not luteinizing hormone-releasing hormone mRNA, levels in male sheep. AB - This study tested the hypothesis that photoperiod affects the ability of testosterone to reduce proopiomelanocortin (POMC) mRNA levels in the arcuate nucleus and luteinizing hormone-releasing hormone (LHRH) mRNA levels in both the preoptic area (POA) or medial basal hypothalamus (MBH). Twenty castrated male sheep were assigned to one of four treatment groups (i): short days (SD; n=5) (ii), short days with testosterone (SD+T; n=5) (iii), long days (LD; n=5), or (iii) long days with testosterone (LD+T; n=5). Blood samples were collected twice weekly for the last 3 weeks of photoperiod treatment and assessed for LH to validate the response to photoperiod. After evaluating LH levels, one animal each from the LD+T and SD+T groups was excluded from the analyses. Mean concentrations of LH were lower (P<0.01) in the LD+T group than in the other treatment groups, which did not differ (P>0.10) from each other. Neither POA nor MBH LHRH mRNA levels were affected (P>0.10) by treatment. Conversely, POMC mRNA levels were suppressed (P<0.01) in the LD+T males compared with the other treatment groups which did not differ (P>0.10) from each other. These observations suggest that photoperiod specific, testosterone-induced alterations in LHRH mRNA levels are not a mechanism whereby testosterone suppresses LHRH release, and that increased beta-endorphin synthesis and release do not mediate testosterone-induced seasonal suppression of LHRH release. PMID- 9725711 TI - Neuroendocrine suppression of female courtship in a wild passerine: corticotropin releasing factor and endogenous opioids. AB - During emergencies in their natural environments, vertebrates initiate coping mechanisms that redirect behavior away from nonessential activities and towards survival. Reproductive behaviors are suppressed. Evidence from field studies on passerine birds shows that this inhibition may not depend on the suppression of gonadal sex steroids, since during the breeding season they remain elevated despite activation of the stress response. We hypothesize that an alternate, central mechanism mediates the inhibition of reproductive behavior during stress in passerines. In this study, we tested the intracerebroventricular effects of endogenous opioids and corticotropin-releasing factor (CRF), neuropeptides implicated in the stress response, on courtship behavior in wild-caught female white-crowned sparrows. Beta-endorphin (beta-EN) significantly inhibited copulation solicitation, an estrogen-dependent courtship display, 30 min after treatment. Naloxone, an opioid antagonist, enhanced the behavior. CRF caused a suppression of solicitation that was reversible by pretreatment with naloxone, suggesting an intermediary role for endogenous opioids in CRF-induced suppression of courtship. The effects of beta-EN and CRF on solicitation appear to be independent of any general effects on locomotor activity. These results support our hypothesis that stress neuropeptides orchestrate coping behaviors in wild birds. PMID- 9725712 TI - Does prolactin mediate induced nest-building behaviour in pseudopregnant gilts treated with PGF2alpha? AB - Nest-building behaviour occurs 6-24 h before parturition in pigs (gestation=116 days). Pseudopregnancy in pigs (induced with oestradiol valerate injections) lasts 50-80 days. We have shown that prostaglandin F2alpha (PG) administration on day 47 of pseudopregnancy induces nest-building and changes to plasma prolactin, oxytocin, cortisol and progesterone similar to those seen before normal parturition. Peripheral prolactin has been proposed as a modulator of nest building. This study assessed nest-building behaviour in prolactin-deprived gilts. Jugular vein catheters were inserted on day 39 of pseudopregnancy and blood samples collected daily from days 40-48. Animals were injected im with either 40 mg bromocriptine in 2 ml 70% ethanol (n=8) or vehicle (n=7) at 17.00 h on day 46 and 09.00 h on day 47 of pseudopregnancy. PG (15 mg Lutalyse: Upjohn) was injected im at 11.00 h on day 47. Blood and behavioural samples were taken from 90 min before PG to 6 h post-PG. Plasma prolactin increased in control but not bromocriptine treated animals following PG (P<0.05). Elevations in oxytocin, cortisol and progesterone (P<0.05) above pre-PG concentrations were also seen, but of these only progesterone showed between group differences [greater (P<0.05) in control gilts on both days 47 and 48]. PG significantly (P<0.05) increased both the rate and proportion of total time spent performing straw/floor-directed behaviours not including foraging (an index of nesting behaviour) in both treatment groups with no significant differences between groups. There were also no significant differences between groups in time spent performing pen fixture directed activities before or after PG. Bromocriptine suppressed the rise in prolactin concentrations after PG without suppressing nest-building behaviour. We conclude that peripheral prolactin is not an essential component of the nest building complex in pigs. PMID- 9725713 TI - Anorectic effect of pituitary adenylate cyclase activating polypeptide (PACAP) in rats: lack of evidence for involvement of hypothalamic neuropeptide gene expression. AB - We investigated the effect of centrally administered pituitary adenylate cyclase activating polypeptide (PACAP) on feeding in rats, and the involvement of hypothalamic neuropeptide gene expression using in situ hybridization. Intracerebroventricular injection of PACAP (1000 pmol/rat) significantly decreased food intake in a dose-dependent manner. In PACAP-treated rats, neuropeptide Y (NPY) mRNA levels in the arcuate nucleus and galanin mRNA levels in the paraventricular nucleus increased, and corticotropin-releasing hormone (CRH) mRNA levels in the paraventricular nucleus decreased. In rats fasted for 72 h, NPY mRNA levels increased, and CRH mRNA levels decreased, but galanin mRNA levels were unchanged. These results indicate that the anorectic function of PACAP is not mediated by NPY or CRH, and that PACAP increases galanin synthesis. PMID- 9725715 TI - Chronic administration of leptin into the lateral ventricle induces sexual maturation in severely food-restricted female rats. AB - In many species, delayed sexual maturation occurs when metabolic conditions are not satisfactory. Recently, leptin was shown to be involved in the regulation of food intake and body mass. Furthermore, leptin administration was shown to advance sexual maturation in mice and to rescue sexual function in adverse metabolic conditions. We examined plasma leptin levels in female rats during development and evaluated the role of leptin on sexual maturation in rats subjected to food restriction. In normal rats, plasma leptin levels were low at day 24 of life, then steadily increased during the juvenile period, reaching 740+/-56 pg/ml at 40 days at time of vaginal opening (VO) and further increasing by day 60 (957+/-73 pg/ml). Food restriction initiated at day 25 strongly impaired this increase, in proportion to the severity of the restriction. With a daily food intake reduced to 7-8 g/day, that permanently prevented VO, plasma leptin levels were very low at day 53 (169+/-67 pg/ml). Following switch to ad libitum feeding, plasma leptin reached high levels within 2 days (1577+/-123 pg/ml), and VO occurred 4 days later. If the severe food restriction was maintained and a central infusion of leptin (10 microg/day) was initiated, a significant decrease in body weight compared with vehicle-infused controls was observed. In these conditions, VO occurred in eight out of the nine leptin treated rats, representing induction of the process of sexual maturation confirmed by increases in ovarian and uterine weights. This induction of sexual maturation exclusively results from a central effect of leptin because no leak of the i.c.v. administered leptin to the general circulation was observed. These data suggest that the rising plasma levels of leptin in the prepubertal period represent a signal to the brain indicating that the young animal is metabolically ready to go through the process of sexual maturation. PMID- 9725714 TI - Ovarian steroid-independent diurnal rhythm in cyclic GMP/nitric oxide efflux in the medial preoptic area: possible role in preovulatory and ovarian steroid induced LH surge. AB - The aim of this study was to evaluate the relationship between cyclic LH hypersecretion and nitric oxide (NO) release in the medial preoptic area (MPOA), the hypothalamic site implicated in induction of LH hypersecretion. The MPOA extracellular cyclic GMP (cGMP) efflux (an index of NO release), was monitored by microdialysis. Quite unexpectedly, we observed a daily afternoon rise in the MPOA cGMP efflux in cycling female rats on proestrus and diestrus II, in ovariectomized (ovx) rats and in ovx rats treated with ovarian steroids to induce the LH surge. The daily rise in cGMP efflux occurred earlier in diestrous and in estradiol benzoate (EB)-treated ovx rats than in ovx rats. Progesterone (P) injection to estrogen-primed ovx rats further advanced the onset of the rise close to the earliest time of rise as seen on proestrus. The afternoon increase in the cGMP efflux in proestrous rats was abolished by pentobarbital treatment that blocked the LH surge. Intracerebroventricular (i.c.v.) injection of 1 H [1,2,4]oxadiazo[4,3-a]quinoxalin-one (ODQ), a selective inhibitor of soluble guanylyl cyclase, suppressed the P-induced LH surge in EB-primed ovx rats, but not basal LH secretion in unprimed ovx rats. Analysis of brain NOS (bNOS) levels in the POA by Western blotting showed that the morning bNOS levels were higher in the POA of EB-treated rats than in unprimed ovx rats. Further, with the exception of ovx rats treated with sequential EB and P treatment, the POA bNOS levels rose significantly in the afternoon in unprimed ovx and EB-treated ovx rats. Collectively, these findings reveal a diurnal rhythm in the MPOA cGMP/NO efflux that is ovarian steroid-independent. Ovarian steroids apparently shift the timing of the afternoon rise in cGMP/NO efflux to synchronize with the activation of steroid-dependent neuronal systems responsible for the LH surge. PMID- 9725716 TI - Stress-induced changes of gene expression in the paraventricular nucleus are enhanced in spontaneously hypertensive rats. AB - Heightened hypothalamic-pituitary-adrenal (HPA) axis responses have been implicated in hypertension in the spontaneously hypertensive rat (SHR), but the exact mechanisms involved are poorly understood. To determine changes in gene expression in SHR in the paraventricular nucleus (PVN), stress-induced accumulation of CRF, CRF type 1 receptor (CRFR-1) genes, and immediate-early genes were examined using in situ hybridization in young (5 weeks old) and adult (12 weeks old) stroke-prone SHR (SHRSP), compared with normotensive Wistar Kyoto (WKY) rats. Restraint stress-induced accumulation of c-fos, jun B, and NGFI-B mRNA, and CRF hnRNA in the PVN was significantly higher in young and adult SHRSP than in WKY rats at 30 min, except for c-fos in young rats. CRFR-1 mRNA expression in the PVN was also significantly higher in adult SHRSP than in WKY rats at 120 min after stress onset. CRF mRNA was increased in response to stress in young SHRSP. The basal CRF mRNA level in the PVN was significantly lower in adult SHRSP than in WKY rats. Young SHRSP exhibit greater ACTH responses to stress without significant changes in plasma corticosterone concentrations. The adult SHRSP exhibited lower baseline concentrations of corticosterone and similar corticosterone response to stress with enhanced secretion of ACTH. Overall, these results demonstrated that stress-induced activation of immediate early genes and CRF gene transcription in the PVN, and ACTH secretion is enhanced in early hypertensive, young, and adult SHRSP, suggesting that they are probably not the result of chronic alterations in blood pressure. The abnormal hypothalamic pituitary response to stress thus appears to be related to the development of hypertension. PMID- 9725717 TI - Bad nursing records will make nursing scholarship suspect. PMID- 9725718 TI - Nurse-patient relationships: the context of nurse prescribing. AB - Nurse prescribing was initiated in the United Kingdom in October 1994 in eight demonstration sites. The evaluation of this extension to the community nurses' role explored both economic and qualitative benefits to patients, carers, nurses and other health care professionals. In this paper the impact of nurse prescribing on patients is explored. Benefits experienced by patients are described along with the difficulties encountered. The patients' views regarding nurses as prescribers are also explored. Data were collected by means of interviews with patients/carers, the focus of which was to evaluate changes associated with nurse prescribing. Patients raised a number of issues associated with their relationship with nurses. Patients valued nurses for both their accessibility and approachability, which led them to discuss health issues which would not otherwise have been brought to the attention of the general practitioner. The arguments which support the incorporation of these qualities into an expanded nursing role are presented. PMID- 9725719 TI - The nature of philosophy of science, theory and knowledge relating to nursing and professionalism. AB - It appears that nursing has devoted an extravagant amount of concentration to the subject of professionalism and professionalization. Consequently, it has created and persists to create some proportion of controversy amid nursing authors, particularly in the Western World at the present time. According to Silva, philosophy, knowledge and theory are intrinsically linked. These notions are important to consider independently and to clarify their relationships, if nursing's knowledge base is to be built on a strong foundation. Perhaps then, if nursing wishes to continue on the trail of professionalization, nurses need to return to and reconsider its foundations and accomplishments. The aim of this discussion paper is to explore the nature of the philosophy of science, knowledge and theory and their interrelationships, with particular reference to professionalization by considering where nursing has come from and consequently the way forward to ascertaining professional status. PMID- 9725720 TI - Fruits without labour: the implications of Friedrich Nietzsche's ideas for the caring professions. AB - Seldom is the work of philosophers invoked by health professionals when examining aspects of care from a philosophical perspective. Instead, students of health care, especially nurses, have been introduced to 'philosophies' which are often superficially examined and poorly understood. This practice fails to develop in students an appreciation of the work of philosophers or to acquire the art of critical thinking. The introduction of models and theories of nursing in the past three decades has alerted nurses to the importance of possessing critical skills in order to identify sound theory and implement good practice. This paper goes beyond mere philosophising and examines aspects of mental health care from the perspectives of one of nineteenth century Europe's most notable philosophers, Friedrich Nietzsche. It argues that understanding his work can enhance one's ability to reflect on nursing practice, as well as bringing a new dimension to how we analyse 'mental health' problems. His work provides many insights into how we can improve our understanding of the effect of mental illness and mental health care on the individual, and how we conceptualise the process of care. This paper provides an overview of his life's work, his impact on the history of ideas and develops some of the more provocative implications of his work for mental health care. PMID- 9725721 TI - Can paternalism be justified in mental health care? AB - Whilst current mental health care provision has made a substantial move towards empowering its users it retains a paternalistic approach with legislation such as 'supervised discharge'. This apparent paradox creates potential dilemmas for the mental health nurse and suggests there is a need for critical analysis of the justification for paternalism within mental health care. This paper discusses the conflict between the ethical principles of autonomy and beneficence which paternalism invokes. On reviewing the ethical theories of deontology and utilitarianism it appears that a prerequisite for autonomy is rationality, the absence of which provides justification of 'weak' paternalism. However, this paper contends that the assessment of rationality has the potential to be subjective and value laden. The use of competency tests can also be problematic by masking the essential ethical dilemma that is intrinsic to such assessment. A case study is used to illustrate the strengths and weaknesses of the competing arguments. This leads to the conclusion that whilst there may be some situations when the use of paternalism can be justified in mental health care, it should be exercised with caution. When there is disagreement between nurse and patient on what is considered to be in the patient's best interests, it should not be assumed that the patient is wrong or irrational. PMID- 9725722 TI - Cut from the right wood: spiritual and ethical pluralism in professional nursing practice. AB - Today, different cultures and contexts of nursing adhere to different codes of ethics. This pluralism may be traced back to differing beliefs about the nature of man and the world, involving different approaches to, and understandings of, spirituality. How significant is this pluralism of beliefs surrounding spirituality for proper nursing practice? I argue that certain introductory nursing textbooks perceive the significance of spirituality for nursing practice as marginal, because of certain assumptions as to what constitutes a proper, or professional, practice. After arguing that such assumptions are problematic, especially from an ethical point of view, I will advance an alternative understanding of professional practice, by drawing upon Alasdair MacIntyre's work. The aim is to give the spiritual dimension of nursing care its rightful place. PMID- 9725723 TI - Fuzzy thinking. AB - Philosophical analysis is increasingly applied to nursing issues. Whereas pre analytical discussions might be sloppy, analysis should bring rigour. This paper explores some mistakes and misconceptions concerning nurse-related reasonings. It uses, as a running theme, a recent paper in the Journal of Advanced Nursing by Rolfe. The mistakes and misleadings to be found in the paper are not uncommon; so the paper's correction quest aims more widely than single author critique. The mistakes and misconceptions could initially appear abstract and without practical outcome; in order to avoid the threatened appearance, a nursing case study is presented, displaying some undesirable consequences for a nurse who is misled by her reading. It is hoped that this paper shows how some accuracy may be well leading. PMID- 9725724 TI - Issues of non-compliance in mental health. AB - This paper explores and questions the ideas and notions in respect of non compliant behaviours. Traditionally, research into this area has focused on identifying individual characteristics from within the adult branch of nursing and within the confines of the medical model of compliance. Little research has investigated the phenomena of non-compliance within the specialty of mental health. It may be suggested that identifying characteristics in this setting is futile. There are many factors determining a client's compliance behaviours. These include gender, social class, race and socio-economic status. Those suffering from mental ill health are often the most vulnerable in society and it may be argued that as such are often coerced into accepting treatment with those who refuse frequently being labelled as non-conformist. The ability of the client to refuse treatment should be based upon informed consent, but is often dependent upon the social prejudice of the health care professional, their misuse of power to bring about compliance behaviours and their inability to act as an effective advocate. Finally, by eliciting information, shared negotiation, planning and improved communication health professionals may improve compliance behaviours based upon the client's needs and wishes. PMID- 9725725 TI - Paradigm dialogues and dogma: finding a place for research, nursing models and reflective practice. AB - Debates within nursing can take the form of useful discussion and critique or verbal conflict generated from inflexible paradigm positions intolerant of differing stances perceived to be of no value. It is suggested that the most profitable process is debate that generates understanding of the strengths and limitations of various tools and techniques and helps identify suitable usage rather than uncritical advocacy or outright rejection. Suggested uses will thus themselves become subject to further debate and practitioners will be encouraged to adopt a use that suits their practice setting and role. The quantitative qualitative research debate, criticisms of the nursing process and nursing models and reflective practice are examined. Dogmatic positions are highlighted and uses that are potentially controversial are identified. PMID- 9725726 TI - The fatigue experience: persons with HIV infection. AB - Fatigue is a frequently reported symptom by persons with HIV infection and one that has an adverse impact on activities of daily living and overall quality of life. Although the concept of fatigue has been studied extensively and discussed in the literature, little is known about the experience of fatigue by persons with HIV infection. A hermeneutic phenomenological study was conducted to investigate the subjective experience of HIV-associated fatigue and to describe the management of fatigue in the context of daily life. In-depth interviews were done with 10 adult patients of an outpatient HIV/AIDS clinic. Thematic analysis identified three concerns that represented the meaning of fatigue for the participants. The first concern was 'Fatigue as a signal of AIDS'. A second concern was 'The mind, the body, the social experience of fatigue'. The third concern was 'Choosing ways to live with fatigue and addiction'. The findings provide insights for nursing practice regarding the subjective meaning of fatigue for patients with HIV and the need for nurses to explore this topic with patients. PMID- 9725727 TI - Health-within-illness: concept development through research and practice. AB - This paper presents the concept of health-within-illness as an opportunity that increases meaningfulness of life through connectedness or relatedness with the environment and/or awareness of self during a state of compromised well-being. The conceptual development evolved through a research programme on women diagnosed with breast cancer and examples from the research are used to illustrate various components of the definition. Evidence from nursing literature and practicing nurses further enhances development of the health-within-illness concept. Although the health-within-illness idea was originally proposed by Moch (1989) and has been referred to in nursing literature, a conceptual definition is not available. Suggestions for further research and practice are included. PMID- 9725728 TI - It hurts most around the heart: a phenomenological exploration of women's anger. AB - Women's anger experience has been poorly understood and insufficiently researched. Yet the emotion of anger is vitally important to women's physical and mental health, and to the quality of their relationships. This phenomenological study was undertaken as an expansion and extension of the Women's Anger Study, the first large survey of the genesis, manifestations and correlates of anger in American women. Although the earlier study contributed to understanding of anger, a deeper examination of the context and meanings of anger experience was sought. Twenty-nine Caucasian women ranging in age from 21 to 66 years were interviewed. Illustrative occupations ranged from homemaker, student, waitress to business executive, professor, and human service professional. Analysis involved thematizing by the researchers independently and within a multidisciplinary phenomenological research group. The thematic structure of women's anger involved a building over time of a confusing mixture of feelings (hurt, frustration, disillusionment) precipitated by a violation of the core values of the self. The precipitant of anger was unfair and/or disrespectful treatment or lack of reciprocity in relationships. When the anger was confined within self, the woman felt helpless and powerless. However, powerlessness was also evident when anger was externalized in an outburst. To the study participants, an angry outburst meant a loss of control rather than ability to achieve control. Women reported a sense of power when using anger to restore justice, respect, and relationship reciprocity. Clinicians can assist women to reflect on their core values and use the power of their anger effectively. Further studies are in progress to examine the relevance of these findings for women of other races and cultural contexts. PMID- 9725729 TI - Critical thinking as an outcome of nursing education. What is it? Why is it important to nursing practice? AB - Critical thinking is currently a highly valued educational outcome throughout the educational spectrum, but particularly so in relation to higher and professional education. International concerns have focused upon citizen's thinking abilities and thus the nations' abilities to function and compete in complex societies and economies. Educationalists are questioning the wisdom of teaching content as opposed to teaching how to think as a means of dealing with relentless information development and change. Nursing education is also embracing the construct critical thinking as a desirable educational outcome. Some commentators refer to the importance of critical thinking in nursing practice as a central component or in some cases a given. This paper intends to examine the background to the construct and address in detail some of the important questions surrounding critical thinking, and its association with competent nursing practice. PMID- 9725730 TI - The place of metaphor and language in exploring nurses' emotional work. AB - An understanding of the importance of language as a symbolic artefact, particularly with reference to metaphor, has been little addressed within the nursing literature. Consequently, the potential richness of people's accounts may be missed. Data from a study which explored the nature of hospice work and nurses' experiences of professional and personal bereavements are used to illustrate the different understanding which an exploration of metaphor can bring to research. Meanings of metaphor and its role in language are initially presented before drawing on data concerned with the emotional aspects of hospice work. Root metaphors of emotion are identified and examples from nurses' accounts indicate how nurses draw on these metaphors to articulate their experiences. The strategies nurses employ to continue working within the hospice culture are identified both practically in their work and metaphorically in the language they use. PMID- 9725731 TI - The power of the word: some post-structural considerations of qualitative approaches in nursing research. AB - This paper takes a post-structural position to discuss some issues related to the use of qualitative research methodologies in nursing research. Two fundamental assumptions which underpin qualitative approaches are explored in relation to how subjectivity is constructed in the process of data collection. These assumptions reveal an ideological position which proposes that reality can be apprehended by capturing the individual's point of view (subjectivity) and that qualitative researchers can directly represent this lived experience in language (linguistic representation). Post-structural approaches challenge these assumptions which are often an uncritically accepted part of the common sense world and therefore taken to be natural. It proposes that these views of subjectivity and language are not natural but rather cultural. There is a recognition of the wider sociopolitical and historical context in which nursing research takes place and a displacement of normative constructions of subjectivity. It provides nursing with the possibility of challenging existing power structures which determine the individual's experiences of health and illness. PMID- 9725732 TI - Collecting and analysing qualitative data: issues raised by the focus group. AB - The focus group has gained considerable popularity as a means of gathering qualitative data in nursing research. This paper examines some of the methodological issues raised by the collection and analysis of focus group data. In respect of data collection, the role of the focus group moderator and the method of recording data are crucial considerations. In particular, the moderator's personal skills and attributes have a considerable influence on the nature and quality of the data gathered. When analysing the data, three principal issues arise. First, it is difficult, and probably misguided, to attempt to infer an attitudinal consensus from focus group data. An apparent conformity of view is an emergent property of the group interaction, not a reflection of individual participants' opinions. Second, measuring strength of opinion from focus group data is problematic. The indicators used to measure attitudes in orthodox survey research are largely inapplicable to the context of focus groups. When comparing data from different focus groups, inferences may be drawn as to the presence of absence of certain views or issues across groups, but not in terms of their relative strength. Third, both methodological and epistemological objections can be raised against attempts to generalize from focus group data. Theoretical generalization is likely to be more feasible than empirical generalization, and if the latter is considered fruitful, it is likely to be of a provisional nature. The effective use of focus groups as a means of gathering qualitative data in nursing research requires due attention to problematic methodological issues such as those explored in this paper. PMID- 9725734 TI - Homophobia: an evolutionary analysis of the concept as applied to nursing. AB - Homophobia is a socially accepted, culturally based belief, which is heavily influenced by an individual's or a community's inherent attitudes, beliefs and values. This conceptual analysis of homophobia has endeavoured to review existing literature on homophobia and subsequently identify and examine the phobic constituents of the concept. References to homophobia are mostly from the 1970 1980 period and there is much unacknowledged conceptual baggage that accompanies the term, which results in restrictive and inappropriate ideas about this concept. This is mainly the consequence of comparisons of homophobia to other phobias, which directly infers fear of homosexuals, while in reality homophobia is more of a biased disgust at homosexuals' lifestyles. This paper attempts to re conceptualize homophobia so that empirical research can begin to test the critical attributes of the concept. This forms the basis for the development of a comprehensive social psychological theory of attitudes towards homosexuals. Such a theory would transcend the unilateral and unidimensional concept of homophobia as a fear and help the understanding of attitudes and feelings towards homosexuals. PMID- 9725733 TI - Collecting patients' views and perceptions of continence services: the development of research instruments. AB - This paper describes one part of a feasibility study carried out in England which examined the costs, quality and effectiveness of continence services in areas with different approaches to provision. It involved the design of instruments to collect the views and perceptions of patients and an investigation of ways to develop a methodology to implement comparative studies. It is the design and piloting of the questionnaires and the way patients responded that forms the focus of this paper. The main sample group recruited for the study were women who had recently sought formal help with urinary incontinence and were likely to receive conservative treatment or management in the community. They were interviewed and asked to complete four questionnaires at two points in time. A smaller sub-group of disabled women, interviewed only once, were included to compare cost profiles for different client groups. The questionnaires which were developed address the impact of urinary incontinence (using a standard scale), the effectiveness of service provision in terms of patients' clinical history, expectations and hoped-for outcomes, service receipt and its cost, and patients' satisfaction with several aspects of service provision. In total 118 women were interviewed, including 28 disabled women. The study generated a set of survey instruments which might be used for a variety of purposes including audit and future research and which could inform purchaser and provider decisions by using patients' perspectives of quality of life outcomes to enhance service development. PMID- 9725735 TI - Trends in community care and patient participation: implications for the roles of informal carers and community nurses in the United Kingdom. AB - This paper argues that the interfaces between formal and informal care-giving are changing as a result of two current trends; the increased scope of home-based nursing care and the emphasis on participation both within nursing and in the wider health and social care arenas. These various changes are explored in relation to the provision of intensive and complex nursing care in the home. It will be argued that the changing interfaces between formal and informal care have important implications for the respective roles of nurses and informal carers which hitherto have been relatively overlooked. These implications urgently need addressing in research, policy and public debate if professional nurses are to provide appropriate help and support to informal carers. PMID- 9725736 TI - A review of current theories of death and dying. AB - Recently, attention has been focused on the care of dying individuals and their families. Although this area of care remains topical for health care professionals, there have been few attempts undertaken to explore critically both the seminal and more recent theories of death and dying in order to extend and develop the theoretical scope and orientation of this field further. The purpose of this paper is therefore twofold: first, to provide a general overview of the current theories of death and dying, and second, to discuss some specific and general contributions and limitations of the current theories. To this end, a review of seminal and more recent theoretical frameworks is presented in this paper. Implications for future research in theory development in the field of death and dying are highlighted. PMID- 9725737 TI - Countertransference in the nurse-patient relationship: a review of the literature. AB - Countertransference is a psychoanalytical concept which, when applied to nursing, refers to the unconscious response of the nurse to the patient. Psychoanalytical concepts such as the unconscious are infrequently mentioned in the nursing literature and have received little research attention. In this paper the literature about the nurse's countertransference is reviewed. In order that the psychodynamic aspects of this phenomenon are more fully appreciated, both the concepts of the unconscious and transference are first described. The nurse's countertransference has many expressions. The literature under review has highlighted the expression of countertransference through physical symptoms, through the nurse's level of involvement with the patient and through the nurse's positive and negative descriptions of patients. The value of recognizing countertransference is universally acknowledged. It is suggested that countertransference in the nurse-patient relationship should be explored further. The knowledge provided would help provide greater insights into the nurse-patient relationship, and ultimately will be reflected in the quality of care which the patient receives. PMID- 9725738 TI - Schizophrenia: all in the mind or locked in the brain? AB - This paper questions whether a natural science language can be transposed either into the care of individuals affected by mental illness or the educational curricula of those preparing to care for them. The importance of biological research into schizophrenia is not denied. However, it is suggested that paradigms which depend upon ownership (of knowledge) may be less worthwhile to schizophrenic people than an approach which rests upon a philosophy of being. In this sense, a consideration of the place of consciousness in investigations into brain function is stated to be relevant: issues of mind and brain are central to discussions about schizophrenia. It is not denied that the laboratory-bench may ultimately unravel genetic susceptibility to schizophrenia. However, a biology of persons - however persuasive its language - can lead, in the case of schizophrenia, to formulations of human defect. Forms of care which proceed from determinism can lead, as they did in the past, to the curtailment of individual aspirations for both carers and patients. PMID- 9725739 TI - Specialist nurse training programme: dealing with asking for organ donation. AB - The issue of cadaveric organ transplantation is by its very nature emotional as it is associated with the very traumatic time of a loved one's death. Making a request for organs needs to be handled very sensitively by health professionals when discussing the issue with a family. Those nurses working in critical care areas are most likely to confront this issue and need to be equipped for dealing with ensuing events. The major challenge for the nurse is to address the concerns with brain death and organ donation in an environment of grief and sadness. Asking for organ consent is the most important element of all and needs to be done in the most sensitive manner, providing appropriate support to the donor family. To facilitate this process specialist training programmes in the nursing curriculum are imperative. Education programmes should incorporate presentations, role play situations and discussions based upon past experiences of organ requests. This would hopefully result in increased rates of donor consent and thereby a reduction in transplant waiting lists. PMID- 9725740 TI - Innovation in the assessment of nursing theory and its evaluation: a team approach. AB - In response to recent national changes in United Kingdom (UK) nurse education (e.g. devolution of assessment, moves to higher education, revision of the aims) and to local concerns (e.g. fairness to students, validity and reliability of written assessments, helping staff with less experience of assessment, student learning) an initiative has been developed at Southampton based on a team approach to marking and moderating. A five-stage evaluation was designed to accompany implementation of the initiative. The evaluation, carried out by a lecturer and an independent educational evaluator, involved both tutors/lecturers and students. Interviews, questionnaires and observation methods were used. Benefits of the initiative and of the particular model of evaluation included: increased knowledge and confidence in the validity and reliability of the marking and moderating process undertaken by tutor-teams; increased fairness to students; in-service tutor training related to student assessment; knowledge that assessment-promoted learning was taking place. A review of the total assessment programme was an unexpected outcome, including a review of the frequency and timing of assessments and of the written guidelines. The five-stage evaluation developed a feeling of involvement and heightened self-knowledge. Curricular understanding also increased; this helped to achieve the initiative as designed and intended. We recommend this model of evaluation; it promotes involvement of all concerned, students as well as staff, and generates valuable process knowledge. It can be used in pre- and post-registration nurse education. PMID- 9725741 TI - The new ward manager: an evaluation of the changing role of the charge nurse. AB - Changes in health service policy and in the structure of nursing have made charge nurses into ward managers with the autonomy and power to improve the quality and effectiveness of patient care. In a climate where clinical effectiveness and quality of care are seen as central to the provision of appropriate health care, this study explores the experiences of one National Health Service trust in implementing this change in the charge nurse's role, and in doing so highlights the importance of effective change management. Nurse managers were interviewed and all ward-based G grade (range A-I) charge nurses within the trust were sent a questionnaire to ascertain: the level of satisfaction with the way the changes had been introduced; whether they were in favour of the changes; and if they had sufficient time, knowledge, resources, preparation and support to enable them to undertake their new role. A random sample of charge nurses were also interviewed. It was found that the majority of charge nurses (61%) were in favour of the development of their role, believing it to be both inevitable and necessary. However, many felt that the change process had been managed ineffectively. There had been insufficient consultation during the change process, preparation and support were perceived as inadequate, the roles of the various protagonists were often unclear, and the lack of supernumerary status led to role conflict and confusion. As a consequence the new ward managers were often unable to fulfil the true potential of this demanding but exciting role. PMID- 9725742 TI - Bridging the research-practice gap: exploring the research cultures of practitioners and managers. AB - To ensure effective utilization of research in nursing more evidence is needed which illuminates the way nurses think about research, the value which they put on it, and how they envisage that it may help or hinder them in their everyday work. This English study aimed to meet these objectives by describing the research culture of practising nurses, health visitors and midwives, and their managers. It rests on two assumptions. Firstly that the reasons why practitioners do, or do not, base their practice on research are complex, and secondly, that interventions to increase research utilization must be grounded in an appreciation of this complex 'whole'. Thus the study took a qualitative approach to exploring: what participants thought and felt about research; the current status of research based practice; and the opportunities and constraints to increasing research based practice. The results confirm the hypothesis that many factors, both individual and organizational affect research utilization. Furthermore, practitioners and managers hold differing perceptions regarding the nature of research, its role, and the opportunities and constraints which effect its dissemination and utilization. The implications of the results for education, policy and practice are discussed. PMID- 9725743 TI - Lifestyle practices and the health promoting environment of hospital nurses. AB - Lifestyle practices and the health promoting environment of hospital nurses This paper examined the lifestyle practices of hospital nurses and the impact of specific interventions in the hospital environment. The perception of nurse as health promoter and as carer of AIDS patients was also examined. A self administered questionnaire was used to collect data at two different time periods. The sample represented 729 nurses (at pre- and post-time periods), both qualified and student nurses. Qualified nurses reported the highest stress levels while student nurses reported more negative lifestyle practices such as smoking, alcohol consumption and drug use. A greater number of current smokers (29%) consumed alcohol and used drugs than non-smokers. The impact of intervention strategies around compliance with smoking policy and work-site walk routes reduced exposure to passive smoking at work for qualified nurses and increased exercise participation for both groups of nurses. Workplace was identified as the main source of stress which included relationships at work and demands of the job. Hospital nurses experiencing high work stress were more likely to use professional support and personal coping (discuss problems with friends/family, have a good cry and eat more) than others. Nurses believed in the importance of health promotion as part of their work; however, qualified nurses felt more confident and gave more health related information than student nurses. Student nurses perceived a lower risk of contacting AIDS through work and a higher concern/worry in caring for AIDS patients than qualified nurses. PMID- 9725744 TI - Feedback: neonatal nurse practitioners. PMID- 9725745 TI - Nicotinic acetylcholine involvement in cognitive function in animals. AB - Nicotinic cholinergic systems are involved with several important aspects of cognitive function including attention, learning and memory. Nicotinic cholinergic receptors are located in many regions of the brain, including areas important for cognitive function such as the hippocampus and frontal cortex. Nicotinic agonists have been found in rodent and non-human primate studies to improve performance on a variety of memory tasks. In a complementary fashion, nicotinic antagonists such as mecamylamine impair working memory function. In humans, similar effects have been seen. Nicotinic agonist treatment can improve attention, learning and memory and nicotinic antagonist treatment can cause deficits. To define the neural substrates of nicotinic involvement in cognitive function, three areas of investigation are underway. 1) Critical neuroanatomic loci for nicotinic effects are beginning to be determined. The hippocampus, frontal cortex and midbrain dopaminergic nuclei have been found to be important sites of action for nicotinic involvement in memory function. 2) Nicotinic receptor subtype involvement in cognitive function is being studied. There has been considerable recent work identifying nicotinic receptor subunit conformation including alpha and beta subunits. Nicotinic receptor subtypes appear to be associated with different functional systems; however, much remains to be done to determine the precise role each subtype plays in terms of cognitive function. 3) Nicotinic interactions with other transmitter systems are being assessed. Nicotine receptors interact in important ways with other systems to affect cognitive functioning, including muscarinic ACh, dopamine, norepinepherine, serotonin, glutamate, and other systems. Nicotinic function in clinical populations and potential for therapeutics has been investigated for Alzheimer's disease, Parkinson's disease, schizophrenia and attention deficit/hyperactivity disorder. Areas which need to receive greater attention are the exact anatomical location and the specific receptor subtypes critically involved in nicotine's effects. In addition, more work needs to be done to develop and determine the efficacy and safety of novel nicotinic ligands for use in the long-term treatment of human cognitive disorders. PMID- 9725746 TI - Behavioral approaches to the assessment of attention in animals. AB - Increasing awareness that disorders of attention may underlie cognitive dysfunctions associated with intoxication and neurodegenerative disease has stimulated research into the neural bases of attention. Because attention comprises a constellation of hypothetical cognitive processes, it can only be inferred from behavior, of either human or non-human subjects, under appropriate experimental conditions. Many behavioral procedures have been proposed for modeling attention in animals, but not all of these procedures have been systematically associated with specific attentional processes. This review endeavors to evaluate critically the construct validity of these procedures (i.e., to determine the degree to which a given procedure assesses a particular process) and to suggest experiments to improve the conceptual links between these procedures and the processes they purport to assess. Five categories of processes have been identified from the animal literature: orienting, expectancy, stimulus differentiation (including stimulus salience, discrimination of a critical stimulus from its context, and selection among stimuli), sustained attention, and parallel processing. The review discusses the strengths and weaknesses of specific behavioral procedures for assessing these categories of attentional processes and, given the conceptual uncertainties involved, it attempts to summarize the present state of knowledge of the pharmacology and neurobiology of attention. PMID- 9725747 TI - Assessment of sustained and divided attention in rats: aspects of validity; [comment]. PMID- 9725748 TI - Nicotine enhances sustained attention in the rat under specific task conditions. AB - Although nicotine has cognitive enhancing effects in both animals and humans, most studies in humans have only shown consistent improvements in sustained attention. Moreover, many studies with smokers have been criticised, since nicotine may simply be relieving withdrawal-induced deficits. The present study investigated the effect of nicotine on sustained attention in drug-naive rats using a five-choice serial reaction time task. Initially, the task was demonstrated to satisfy some of the criteria for the construct validity of a vigilance task: reducing signal length and either increasing or decreasing the inter-trial interval significantly (P<0.05) impaired performance. Whether nicotine (0.05-0.4 mg/kg, SC) reversed the deficits induced by a signal length of 0.25 s (weak signal) or an inter-trial-interval of either 20 s (low event rate) or 1 s (high event rate) was assessed. Nicotine (0.15 mg/kg) improved accuracy and decreased omission errors under low event rate conditions only. However, nicotine (0.05/0.15 mg/kg) improved reaction time and increased anticipatory responses under both weak signal and low event rate conditions. There was no effect of nicotine on performance under high event rate conditions. Under the low event rate condition, nicotine enhanced the ability of rats to maintain attention (i.e. accuracy) throughout a session. These findings suggest (i) that nicotine's effect on attention depends upon task characteristics; (ii) these effects on attention may reflect self-reports by smokers that nicotine aids concentration, particularly in stressful situations, and (iii) nicotinic agonists may have therapeutic benefits in patient populations suffering from attentional deficits. PMID- 9725750 TI - The pharmacology of impulsive behaviour in rats II: the effects of amphetamine, haloperidol, imipramine, chlordiazepoxide and other drugs on fixed consecutive number schedules (FCN 8 and FCN 32). AB - The effects of drugs on one aspect of impulsive behaviour were evaluated using a schedule in which rats were trained to complete a fixed consecutive number of responses on one of two levers before pressing the second to obtain a reinforcer (FCN). Terminating the chain before completing the FCN resulted in the omission of the food, and can be considered an impulsive decision. Two groups of food deprived rats were trained to press either 8 or 32 times on the left lever (FCN lever) of a two lever operant chamber before pressing the right lever (Reinforcement lever) to deliver a food pellet. Responding on the Reinforcement lever before completion of the sequence resulted in a short time-out and the rat had to begin the sequence again. After responding had stabilised, the rats were treated with a range of doses of a number of drugs. Impulsivity was assessed by several measures, including the mean chain length and the proportion of chains terminating in food delivery, and the distribution of chain lengths was analysed. The efficiency of the rats was similar under both FCN 8 and FCN 32, although it was more difficult to maintain a consistent baseline under FCN 32. Under the FCN 8 schedule, significant decreases in chain length were obtained with d amphetamine (0.8-2.4 mg/kg), haloperidol (0.1 mg/kg), ethanol (1 and 3 g/kg) and chlordiazepoxide (10.0 mg/kg), and there were alterations in other measures consistent with an increase in impulsivity. Imipramine (1-10 mg/kg), citalopram (1-10 mg/kg) and metergoline (0.3-3.0 mg/kg) had no effect on mean chain length, although the first two drugs shifted the chain length distribution to the left. d Amphetamine (0.4-1.2 mg/kg) and PCPA (100 mg/kg) reduced chain length and had other effects consistent with increased impulsivity under FCN 32 schedule, whereas imipramine had little, and citalopram no, effect. Taken generally, effect of the active drugs was relatively non-specific, including both a reduction in response rate and alterations in choice measures proposed to reflect an increase in impulsivity. Detailed analysis of the effect of amphetamine revealed that three processes were at work: chain shortening, an increased preference for the lever most closely associated with food delivery, and a gradual shift in the control over responding from the response sequence (pattern) to the individual lever press (act). PMID- 9725749 TI - Sabcomeline (SB-202026), a functionally selective M1 receptor partial agonist, reverses delay-induced deficits in the T-maze. AB - Sabcomeline, (SB-202026 [R-(Z)-alpha-(methoxyimino)-1 -azabicyclo [2.2.2] octane 3-acetonitrile]), a functionally selective muscarinic M1 receptor partial agonist, was tested in rats trained to perform a delayed, reinforced alternation task in a T maze, a test of short-term spatial memory. For comparison the cholinesterase inhibitor tacrine (THA-9-amino- 1,2,3,4-tetrahydroaminoacridine) and the non-selective muscarinic receptor agonist RS86 (2-ethyl-8-methyl-2,8 diazospiro [4.5]-decane-1,3-dione hydrobromide) were also tested and all three compounds were also compared using a conditioned taste aversion (CTA) task. Sabcomeline (0.001-1.0 mg/kg i.p.) significantly reversed the T-maze choice accuracy deficit induced by a 20-s delay at 0.03 and 0.1 mg/kg. RS86 (0.1-3.0 mg/kg i.p.) reversed the deficit at 1.0 mg/kg and THA (0.1-3.0 mg/kg i.p.) had no effect at any dose. All three compounds induced conditioned taste aversion with minimum effective doses (MED) of 0.3, 1.0 and 3.0 mg/kg, respectively. The results show that sabcomeline reverses delay induced deficits in T-maze choice accuracy in a rewarded alternation task at doses approximately 10 times lower than those required to induce conditioned taste aversion. RS86 was equipotent in both tests. These data support the findings of clinical studies which have shown that SB-202026 provides significant symptomatic improvement in patients with probable Alzheimer's disease at doses which do not induce cholinergic side effects. PMID- 9725751 TI - The pharmacology of impulsive behaviour in rats III: the effects of amphetamine, haloperidol, imipramine, chlordiazepoxide and ethanol on a paced fixed consecutive number schedule. AB - The behavioural trait of impulsivity may be made up of different components, including rapid decision making, intolerance to the delay of reward and a tendency to terminate chains of responses prematurely. It has been proposed to measure the last of these in rats using fixed consecutive number (FCN) schedules. The present study uses a modified version of the FCN procedure in which responding was paced by retracting the response lever for short periods between presses. In this way, the experimenter could control the maximum rate of responding. The procedure was made up of two components based on an FCN 8 schedule of food reinforcement. In the Fast component, lever presses were spaced by a minimum of 2 s and in the Slow component by a minimum of 5 s. The average chain length was significantly shorter, and the rats were less efficient in the Slow component. Five drugs were tested on this baseline, imipramine (1.0-10.0 mg/kg), ethanol (300-3000 mg/kg administered PO), haloperidol (0.01-0.1 mg/kg), chlordiazepoxide ( 1.0-10.0 mg/kg) and d-amphetamine (0.2-0.8 mg/kg). All the drugs reduced responding at the highest dose, but imipramine was different from the others in that it increased the average number of responses in the chain and produced a shift in the chain length distribution to the right, possibly reflecting a reduction in impulsivity. The other four drugs reduced chain length at the highest dose, although in the case of ethanol this effect was very small and, unlike the other three drugs, did not result in a shift in the distribution to the left. The paced FCN procedure can differentiate the effects of different drugs on one aspect of impulsivity, and is likely to be a useful procedure for further study of this aspect of behaviour. PMID- 9725753 TI - Triazolam-induced changes in alcoholic thought processes. AB - This study was designed to examine and contrast cognitive effects (explicit memory and access to semantic knowledge) of the benzodiazepine Halcion (triazolam) in ten normal volunteers and ten cognitively un-impaired detoxified alcoholics. The two groups were indistinguishable from one another under placebo conditions on all measures of cognitive functioning. Under Halcion test conditions (0.375 mg p.o.), both groups were about equally impaired in their recall of to-be-remembered information. However, alcoholics, were more likely to recall information that they were not asked to remember (intrusion errors) on all measures of explicit remembering. Alcoholics also generated relatively uncommon (low frequency) responses from semantic memory, rather than common, categorically related associations in response to stimuli such as types of vegetables, flowers, and fruit following the administration of Halcion, but were not different from normal volunteers in the types of responses generated under placebo conditions. These findings suggest that a drug challenge that simulates many of the effects of acute alcohol administration induces alcoholics to think and remember differently (qualitatively) from normal volunteers. PMID- 9725752 TI - Memory for emotional events: differential effects of centrally versus peripherally acting beta-blocking agents. AB - Substantial evidence from animal research indicates that enhanced memory associated with emotional experiences involves activation of the beta-adrenergic system. This hypothesis is further supported by the finding in human subjects that blockade of beta-adrenergic receptors with propranolol selectively reduced memory for emotional events. In the present study, we compared the effects of propranolol, a lipid soluble drug which crosses the blood-brain barrier easily, with those of nadolol, a water soluble drug which crosses the blood-brain barrier to a considerably lesser extent, to determine whether the effect involved peripheral or central beta-adrenergic receptors. The effects of these drugs, taken before subjects watched a slide show that was either emotionally arousing or relatively neutral in content, were tested 1 week later with a surprise memory test. Consistent with previous results, propranolol impaired memory (recall and recognition) in the subjects who saw the emotional version of the slide show. In contrast, nadolol did not impair memory of the emotional slide show. These results indicate that the blockade of central beta-adrenergic receptors is responsible for the reduction in storage of emotional events. The results support the view that memory of a mild emotional event involves activation of central, but not necessarily peripheral beta-adrenergic receptors. PMID- 9725754 TI - Confidence level and feeling of knowing for episodic and semantic memory: an investigation of lorazepam effects on metamemory. AB - The effects of lorazepam (0.026 or 0.038 mg/kg), a benzodiazepine, and of a placebo on metamemory, i.e. knowledge about one's own memory capabilities, were investigated in 36 healthy volunteers. Accuracy of confidence levels (CL) in the correctness of recalled answers and accuracy of feeling of knowing (FOK) the answers when recall fails were measured using a sentence memory task assessing episodic memory and a task consisting of general information questions and assessing semantic memory. Lorazepam impaired episodic memory. Unexpectedly, it also impaired performance in both the recall and recognition phases of the task assessing semantic memory, suggesting that it decreased the ability to distinguish between correct and incorrect information. In episodic memory, lorazepam 0.038 mg/kg-treated subjects exhibited an impaired CL accuracy, compared to placebo-treated subjects, and their FOK accuracy was at chance. In semantic memory, their overall CL and FOK accuracy was apparently spared. However, these subjects selectively overestimated their CL judgements for incorrect answers; moreover, secondary analyses showed that FOK accuracy for a subset of low-accuracy items was virtually nil. These results suggest that lorazepam impairs metamemory for both episodic and semantic memory. PMID- 9725755 TI - Lorazepam but not diazepam impairs identification of pictures on the basis of specific contour fragments. AB - We tested the effects of lorazepam 0.038 mg/kg and diazepam 0.3 mg/kg on the identification of pictures of everyday objects that were specifically modified to examine the role of different parts of the external contour. By pressing the space bar of a computer keyboard, observers could add 1% of the total contour of each picture until it was recognized. Identification thresholds were measured in three display conditions, depending on where along the contour the addition of contour pixels started. In the minima condition, stimuli were initially displayed with only minima (i.e., locations along the contour where negative curvature is strongest); all parts with negative curvature were then built up gradually from the minima and only later on were the fragments with positive curvature shown until the contour became closed at the maxima (i.e., locations where positive curvature is strongest). In the maxima condition, initially only the maxima were displayed, with all positive contour built up first and then the negative curvature until the minima were reached. In the inflections condition, the points along the contour shown first were inflections (i.e., points where curvature is locally zero because the sign of curvature changes there) and contour was built up by adding parts of positive and negative curvature at both sides of each inflection until the extrema (minima and maxima) were reached to close the contour of the picture. In general, picture identification was more difficult (i.e., a larger portion of the contour was required) in the minima condition than in the maxima and the inflections conditions. The diazepam group did not differ significantly from the placebo group, while the lorazepam group had significantly lower performance in all three display conditions. Results are discussed in relation to previous research showing impaired perceptual integration and impaired implicit memory under lorazepam influence. PMID- 9725756 TI - Proactive interference and temporal context encoding after diazepam intake. AB - Two experiments were designed to test whether the memory impairment induced by benzodiazepines (BZDs) is due to impaired memory for temporal context. In both experiments, subjects were administered either diazepam (15 mg oral) or placebo, and a standard BZD impairment on prose recall as well as a decreased subjective arousal was found. Key tasks to explore temporal context memory were an A-B A-C proactive interference paradigm and a list discrimination task. Initial learning of both groups on these tasks was broadly matched. In experiment 1, diazepam did not increase susceptibility to proactive interference using semantically related words. However, in experiment 2, using unrelated word pairs, diazepam markedly increased the number of prior list intrusions. Furthermore, after diazepam intake, subjects were clearly impaired in learning unrelated word pairs. Subjects after diazepam intake were not impaired in the list discrimination task. We conclude that (1) diazepam impairs the forming of new associations, whether this is the formation of links between two or more targets or between targets and context, (2) a temporal context encoding deficit cannot account for a broader diazepam-induced memory impairment. PMID- 9725757 TI - A further examination of the time-dependent effects of oxazepam and lorazepam on implicit and explicit memory. AB - Until recently, research indicated that all benzodiazepines impair explicit memory, while only lorazepam impairs priming. Stewart and associates provided preliminary data which indicated that both oxazepam and lorazepam may impair implicit memory, but in a time-dependent fashion. The present study was designed to replicate Stewart et al.'s findings after overcoming several limitations of the original study. Thirty subjects were administered an acute dose of lorazepam (2 mg), oxazepam (30 mg) or a placebo and were tested with an implicit (word-stem completion) test and an explicit (cued recall) test. However, subjects were only tested at 170 min post-drug (close to oxazepam's theoretical peak concentration) to rule out the possible "explicit memory contamination" explanation of the Stewart et al. implicit memory findings. Consistent with previous research, both drugs impaired explicit memory relative to placebo. Also, both lorazepam and oxazepam impaired priming performance, supporting the "time-dependence" interpretation of the Stewart et al. findings. The results also indicate that episodic memory is impaired by both benzodiazepines in a time-dependent fashion even when the research methodology used involves everyday memory demands. PMID- 9725759 TI - Effortful processing is a requirement for nicotine-induced improvements in memory. AB - We report two studies examining the effects of nicotine on memory in minimally deprived smokers. In experiment 1, semantically related words were recalled significantly better than unrelated words following nicotine, even when volunteers were explicitly instructed to target the unrelated word set for recall. Experiment 2 examined the effect of nicotine on two different types of lexical association: association by joint category membership (semantically related items), and association by derived meaning ("encapsulated" word pairs). Nicotine-induced improvements in recall were observed only for category associates and not for encapsulated word pairs. This implies that explicit, effortful processing of material in the presence of nicotine is necessary for improved recall performance to be observed. PMID- 9725758 TI - Dual task performance after diazepam intake: can resource depletion explain the benzodiazepine-induced amnesia? AB - It was tested whether a depletion in resources can account for the benzodiazepine induced memory impairment. In two experiments, it was examined whether dividing attention had a disproportionately detrimental effect on learning semantically related and unrelated word pairs after diazepam intake. Word pairs had to be learned in both a single task condition and while performing a visual discrimination task concurrently (dual task condition). Moreover, the complexity of the visual discrimination task was manipulated systematically. Diazepam (15 mg, orally) or placebo was administered in a double-blind, between-subjects design. Subjects after diazepam intake were clearly impaired in learning unrelated word pairs, but not in learning related word pairs. Dividing attention in the dual task condition was associated with a reduction in learning unrelated word pairs, but this was not disproportionately reduced after diazepam intake. Moreover, the magnitude of resource depletion did not correlate with the severity of the diazepam-induced memory impairment. In general, the pattern of results does not support the hypothesis that a depletion of resources can explain the benzodiazepine-induced memory impairment. PMID- 9725760 TI - The influence of acetylsalicylic acid on cognitive processing: an event-related potentials study. AB - The central effects of acetylsalicylic acid (ASA) are discussed controversially. In animal models, it has been shown that ASA can interact with the central serotonergic and catecholaminergic neuronal system. However, the relevance of this interaction for humans is still unknown. We performed a study on the influence of ASA on central cognitive processing. In 25 healthy subjects (age 21 56 years), visually evoked event-related potentials (ERP) and reaction time under IV ASA medication were recorded. ERP were evoked by an oddball paradigm. As compared to placebo, ASA decreased the latency of the P3 component significantly in a time interval of 20-40 min after administration. The latency of the N2 component was significantly decreased about 25 min after administration; the latency of the exogenous P2 component was not influenced by ASA. The mean choice reaction time was significantly decreased by ASA 35 min after administration. At this time point, there was a significant correlation between decrease in reaction time and increase in ASA plasma level. The data show that IV administration of ASA has an accelerating effect on the endogenous components of visual ERP and on reaction time. This finding suggests that ASA can influence central cognitive processing, possibly by ASA induced changes of neurotransmitters. Since serotonin can be released by ASA and serotonin release leads to a decrease of ERP latencies. we assume that ASA most likely influences cognitive processing via the central serotonergic transmitter system. PMID- 9725761 TI - FISHing for complements on chromosome 12p. PMID- 9725762 TI - Virtues of being faithful: can we limit the genetic variation in human immunodeficiency virus? AB - Human immunodeficiency virus (HIV) infections are characterized by a high degree of viral variation. The genetic variation is thought to be a combined effect of a high error rate of reverse transcriptase (RT), viral genomic recombination, the selection forces of the human immune system, the requirement for growth in multiple cell types during pathogenesis, and persistent immune activation associated with HIV disease. This hypermutability gives the virus an ability to escape mechanisms of innate immune surveillance and therapeutic interventions. Indeed, HIV variants that are resistant to drugs that antagonize both the HIV protease and RT enzymes are well described. Furthermore, there are seemingly no procedures to restrict this disarming property of HIV to mutate rapidly. Recently we have shown that some of the drug-resistant RTs display an increased in vitro polymerase fidelity. The question is whether this finding will stimulate new approaches that will not only help the immune system to deal with the virus more efficiently but also to reduce or delay resistance to various classes of anti-HIV drugs. The pros and cons of this concept and the influence of viral replication rates and viral fitness on HIV variability are discussed. PMID- 9725763 TI - Early growth response factor 1: a pleiotropic mediator of inducible gene expression. AB - The zinc-finger transcription factor, early growth response factor-1 (Egr-1) is an immediate-early gene product inducibly expressed in response to diverse stimuli. This review examines the emerging role of Egr-1 in vascular pathobiology. PMID- 9725764 TI - Clinical implications of mast cell-bacteria interaction. AB - Mast cells are traditionally known for mediating allergic reactions. In addition, these cells have been implicated in the pathogenesis of a variety of clinical conditions such as atopic and contact dermatitis, bullous pemphigoid, fibrotic lung disease, neurofibromatosis, psoriasis, scleroderma, rheumatoid arthritis, interstitial cystitis, ulcerative colitis, and Crohn's disease, but their role in host defense was an enigma until recently. Owing to the strategic location of mast cells at the host environment interface, their role in bacterial infections has been studied by a number of investigators. Latest reports show that mast cells have an ability to modulate the host's innate immune response to infectious agents. This review discusses the clinical implications of mast cell-bacteria interactions. PMID- 9725765 TI - Understanding, predicting and controlling the emergence of drug-resistant tuberculosis: a theoretical framework. AB - The high prevalence of tuberculosis in developing countries and the recent resurgence of tuberculosis in many developed countries suggests that current control strategies are suboptimal. The increase in drug-resistant cases exacerbates the control problems. Currently employed epidemic control strategies are not devised on the basis of a theoretical understanding of the transmission dynamics of Mycobacterium tuberculosis. We describe and discuss a theoretical framework based upon mathematical transmission models that can be used for understanding, predicting, and controlling tuberculosis epidemics. We illustrate how the theoretical framework can be used to predict the temporal dynamics of the emergence of drug resistance, to predict the epidemiological consequences of epidemic control strategies in developing and developed countries, and to design epidemic control strategies. PMID- 9725766 TI - Molecular and cell biological aspects of neuroendocrine tumors of the gastroenteropancreatic system. AB - Neurons and neuroendocrine cells share a variety of common characteristics. Cell and molecular biological studies in recent years have improved our understanding of physiological and pathophysiological processes such as cellular growth, adhesion, and secretion of neuroendocrine cells. Here we review current findings from the area of basic research and from current clinical research relevant to improving the diagnosis and therapy of neuroendocrine tumors of the gastroenteropancreatic system. PMID- 9725767 TI - Triple-color FISH analysis of 12p amplification in testicular germ-cell tumors using 12p band-specific painting probes. AB - Forty-nine surgical specimens and nine germ cell tumor lines were analyzed by triple-color FISH using microdissected probes for the cytogenetic bands of chromosome arm 12p (12p11.2, p12, and p13). FISH analysis demonstrated amplification of material from all three bands in all tumors. This amplification was in the form of increased copy number of 12p or i(12p) and/or 12p amplified regions (AMP12p). The number of copies of 12p was variable (4-11 copies) from case to case but tended to remain relatively constant in all clones of the same tumor, even when the amplification took the form of an amplified region composed of 12p material. In tumors with multiple clones, i(12p) and AMP12p were never found in the same cell. No correlation was found between 12p copy number and tumor type. We describe, for the first time, a relative overrepresentation of 12p13 or 12p12-p13 regions in six tumors (two surgical samples and four cell lines), either as "partial 12p" (five cases) or within a 12p amplified region (one case). The ubiquitous amplification of all three 12p bands in germ-cell tumors supports the hypothesis that 12p harbors more than one gene important for oncogenesis of adult male germ-cell tumors, and that these genes may be located in different areas of 12p. PMID- 9725768 TI - Developmental study on reduction and kinks of the tail in a new mutant knotty tail mouse. AB - The knotty-tail (knt/knt) mouse has a short and knotty tail. The tail deformity is caused by a decrease in the number of caudal vertebrae and a deformity of them in the distal part of the tail. The objective of the study was to determine how reduction and kinks of the tail region were formed during secondary body formation. By day 12.0 pc, the somitogenesis of knt/knt embryos was completed; the number of caudal somites more or less agreed with those of the caudal vertebrae in knt/knt mice and were similar to those of knt/+ embryos. On the other hand, the somitogenesis of knt/+ embryos continued up to day 12.5 pc. The somites below about the sixth caudal somite were wedge-shaped with a dorsal apex in knt/knt embryos. The location of abnormal somites also corresponded well to that of deformed caudal vertebrae. Abnormal somitogenesis was always preceded by abnormalities in the presomitic region. Under gross observation, this could be seen to become markedly thickened, and histologically its dorsoventral diameter increased in the transverse plane on days 10.5-12.0 pc. In the mesenchyme there was often obvious cell death at the boundary of the unsegmented area and the tail bud after day 10.5 pc. These results suggested that the shortness of tail was primarily caused by the agenesis of distal caudal vertebrae following the agenesis of distal caudal somites, and partly by the disappearance of the presomitic part due to cell death, while the tail kinks were caused by the deformation of each caudal vertebra following disturbances of the caudal somites. Also, it is highly probable that the prominent cell death at the boundary of the unsegmented area and the tail bud may involve a defect or deformity of somites in this mutant. PMID- 9725769 TI - Ultracytochemistry of 3beta-hydroxysteroid dehydrogenase in Leydig cell precursors and vascular endothelial cells of the postnatal rat testis. AB - In the biosynthesis of steroid hormones 3beta-hydroxysteroid dehydrogenase (3beta HSD) is a key enzyme. The present report describes the subcellular localization of the enzyme in the fetal-type Leydig cells, the fibroblast-like precursors of adult-type Leydig cells and in endothelial cells of interstitial capillaries. Histochemical methods for light microscopy and ultracytochemical methods for electron microscopy were used on rat testes of postnatal day 15. 3beta-HSD reactivity was located at subcellular levels by means of the ferricyanide method. A specific, distinct localization of reaction product in the form of copper ferrocyanide precipitates was observed on the membranes of the smooth endoplasmic reticulum not only in the fetal-type Leydig cells and the fibroblast-like precursors of adult-type Leydig cells, but also focally in the endothelial cells of interstitial blood capillaries. Topographically, the 3beta-HSD-positive precursors were most often found in the outer layer of the boundary tissue and surrounding interstitial blood vessels. The capillaries with 3beta-HSD-positive endothelial cells were usually located in the vicinity of 3beta-HSD-positive Leydig cells. For the first time, 3beta-HSD has been located at the subcellular level in precursors of adult-type Leydig cells and focally in capillary endothelial cells associated with them. Due to the close association between 3beta-HSD-positive vascular endothelial cells and Leydig cells a paracrine relationship between the two cell types may be involved in the acute regulation of steroidogenesis by blood-borne luteinizing hormone. PMID- 9725770 TI - Reactive gliosis of immature Bergmann glia and microglial cell activation in response to cell death of granule cell precursors induced by methylazoxymethanol treatment in developing rat cerebellum. AB - The morphology, organization and expression of proliferating cell nuclear antigen (PCNA) and the cytoskeletal proteins vimentin and GFAP in immature Bergmann glial cells were studied after a developmental injury induced by a single dose of the cytotoxic agent methylazoxymethanol (MAM) administered on postnatal day 5. This drug, which produces cell death of cerebellar granule cell precursors, did not induce apoptosis in Bergmann glial cells, which are in a proliferative stage. After MAM treatment, PCNA staining showed a severe depletion of PCNA-positive granule cell precursors, whereas PCNA-positive Bergmann glial nuclei in the Purkinje cell layer were preserved. Moreover, the quantitative analysis revealed an increase in the density of both Purkinje cells and PCNA-positive Bergmann glial cells per mm of Purkinje cell layer in MAM-treated rats relative to age matched controls, but the numerical ratio between these two cell populations remains invariable after MAM treatment. Vimentin and GFAP immunocytochemistry revealed a reinforcement of the Bergmann glial palisade with overexpression of both proteins and thicker immunoreactive glial processes in MAM-treated rats. At the ultrastructural level, Bergmann glial processes closely associated with dying cells in different stages of apoptosis were observed. Frequently, these processes enclosed dying cells in extracellular compartments. Furthermore, phagosomes containing apoptotic bodies were found in Bergmann fibers of MAM-treated rats. These data indicate that the cell death of granule cell precursors triggers a reactive response in immature Bergmann glia. We suggest that this response reflects the plasticity of Bergmann glia to control the neuronal microenvironment in the maturing molecular layer, protecting healthy cells against the potentially harmful contents of dying cells. In situ labeling of cell death with the TUNEL method revealed that the cell death of granule cell precursors is of the apoptotic type. The participation of ameboid microglial cells in the phagocytosis of apoptotic cells was shown with tomato lectin histochemistry and ultrastructural analysis. Moreover, the presence of mitosis in this microglial population demonstrates its proliferative activity in regions of extensive cell death. PMID- 9725771 TI - New indirect pathways subserving the pupillary light reflex: projections of the accessory oculomotor nuclei and the periaqueductal gray to the Edinger-Westphal nucleus and the thoracic spinal cord in rats. AB - The pupillary light reflex (PLR) is under the control of retinal ganglion cells projecting to the olivary pretectal nucleus (OPN). The OPN has a major projection to the Edinger-Westphal (EW) nucleus, which exerts its parasympathetic action on the iris musculature via the ciliary ganglion. The accessory oculomotor nuclei (AON) and the periaqueductal gray (PAG) receive input from the OPN and influence the PLR. The present study in rats aimed to elucidate the possible projections from the AON and PAG to the EW nucleus. The anterograde tracer Phaseolus vulgaris leucoagglutinin (PHA-L) was iontophoretically injected into the interstitial nucleus of Cajal (INC), the nucleus of the posterior commissure (NPC), the nucleus of Darkschewitsch (ND) and the rostral part of the PAG. The projections were studied at the light and electron microscopic level. The INC, NPC and ND have small projections to the EW nucleus, whereas the rostral part of the PAG densely projects to the EW nucleus. Without exception INC, NPC, ND and PAG varicosities are presynaptic to dendritic profiles in the EW nucleus and contain electron dense mitochondria, round vesicles and make asymmetric synaptic contacts. In addition the ND and PAG project to the thoracic level of the spinal cord. The fibres are presynaptic to dendritic profiles and contain electron dense mitochondria, round vesicles and make asymmetric synaptic contacts. The present observations allow the conclusion that the parasympathetic preganglionic neurons in the EW nucleus are not only controlled by the OPN-EW pathway but also by indirect pathways running via the AON and PAG. Moreover light-sensitive information is also transferred via an OPN-PAG-spinal cord pathway to the sympathetic superior cervical ganglion (SCG) that innervates the iris, suggesting that the PAG may have an integrative function in the sympathetic and parasympathetic control of the PLR. PMID- 9725772 TI - Epidermal growth factor, vascular endothelial growth factor and progesterone promote placental development in rat whole-embryo culture. AB - The development and survival of rat embryos in whole-embryo culture is limited by the lack of any maternal blood circulation in a purely fetal placenta. If the resulting placental insufficiency could be overcome for some time by an increase of the placental exchange area, a prolonged culture period would result and facilitate the development of embryos. In the present study, several attempts to stimulate proliferation and growth of the fetal placenta were made by the addition of vascular endothelial growth factor (VEGF), epidermal growth factor (EGF) and progesterone to the culture medium. Rat embryos were routinely explanted with their embryonic membranes at 10.5 days of gestation. Decidua, parietal yolk sac, Reichert's membrane and the layer of superficial trophoblastic giant cells were removed. The explants were cultured and gassed continuously for 24 h in rotating plastic tubes containing rat serum, diluted to 50% with modified COON's F12 medium. Either of the two growth factors or progesterone were added to each culture tube and a control group was cultured without any factor. After the addition of each of these factors the stimulatory effect on placental growth was assessed by morphometric evaluation of several placental parameters from semithin cross-sections: On adding each of the factors the whole cross-sectional area of the placenta significantly increased, as did the area of the fetal placental mesenchyme. VEGF also increased the area of the trophoblast, and the area of the blood vessels enclosed within the trophoblast, by an average of 9.4% and 23.6%, respectively. Thus, VEGF treatment resulted in a measurable extension of the exchange area of the fetal placenta. PMID- 9725773 TI - Differences in genetic backgrounds affecting gonadal differentiation between two local populations of the Japanese wrinkled frog (Rana rugosa). AB - Gonadal differentiation in two local populations - Hamakita and Hiroshima - of the Japanese wrinkled frog (Rana rugosa) was examined from hatching to the end of metamorphosis. The Hamakita male has X and Y chromosomes distinguishable morphologically from each other, while X and Y chromosomes in the Hiroshima male cannot be distinguished morphologically. In the two populations, primordial gonads differentiated into testes or ovaries by stage I of Taylor and Kollros, and larvae at stage XXV - the end of metamorphosis - possessed definitive testes or ovaries. However, immature testes at stages II-III in the Hiroshima population showed a higher incidence of testes with ovarian characteristics - meiotic figures and gonadal cavities - than those in the Hamakita population. In addition, proliferative activities in the testicular cells of the immature testes in the Hiroshima population were very low. On the other hand, there were no remarkable differences in the histological processes of ovary development between the two populations. Intraspecific hybridization between females of the Hamakita population and males of the Hiroshima population retarded testis differentiation after stage XV, while the reciprocal hybridization showed normal gonadal sex differentiation and development. These observations suggest that R. rugosa, possessing heteromorphic sex chromosomes in the male, establishes firm gonochorism, where the feminization involves the suppression of testis differentiation, and that masculinization is performed by resistance to this suppression as well as active proliferation of testicular cells. PMID- 9725774 TI - Prenatal development of the vestibular ganglion and vestibulocerebellar fibres in the rat. AB - We have used carbocyanine dye tracing techniques in conjunction with photoconversion and electronmicroscopy to examine the prenatal development of the central and peripheral processes of those vestibular ganglion cells projecting to the cerebellum. Developmental changes in the number of vestibular ganglion cells were assessed in paraffin-embedded material by nucleolar counting. In agreement with the results of parvalbumin staining, afferents to the cerebellum from the vestibular ganglion pursued a superficial course during early fetal life (E13 to E15). From E16 to E19, this superficial position was progressively lost and vestibulocerebellar fibres were seen to be directed towards the ventricular surface (prospective posterior/inferior vermis). The change in the course of vestibular afferents to the cerebellum coincided with a profound reduction in the number of ganglion cells which could be retrogradely 1a-belled from the cerebellar anlage (mean+/-SD: E16-2040+/-1130; E19-510+/-440). During that same period the total number of vestibular ganglion cells rose to peak at a mean of 9200 at E19, although there was a subsequent decline to an average of 4660 at P0. This population size was maintained through to adult life (4600). We also examined the development of connections between the vestibular ganglion and the vestibular apparatus. Peripheral processes of vestibular ganglion cells invaded the macula utricle and saccule and cristae of the semicircular canals from E13. We found that the peripheral vestibular ganglion cell processes themselves did not show any significant morphological changes from E16 to E21, but the sensory epithelium itself adopts a mature pseudostratified appearance by E21. This suggests that the loss of vestibular ganglion cells from E19 to birth is not related to major morphological changes in the peripheral axons, at least as revealed by carbocyanine dye labelling of these from the cerebellum, but may be associated with differentiation of the sensory epithelium to the mature pseudostratified form. Electronmicroscopy of photoconverted vestibulocerebellar fibres showed that at E14 these afferents were grouped in tight bundles of up to 20 axons. No particular association with the superficially placed external granular layer cells was found at that age. By E16 photoconverted vestibulocerebellar axons were no longer as tightly bundled and could be seen coursing more ventrally through the cerebellar anlage. The findings indicate that vestibulocerebellar fibres are not likely to physically facilitate external granular layer migration, since they do not attain a particularly close structural association with those cells. The observed developmental changes in the number of vestibular ganglion cells projecting to the cerebellum and the total number of vestibular ganglion cells suggests that changes in the course of vestibulocerebellar fibres are associated at first with retraction of cerebellar afferents, and subsequently with developmental cell death in the ganglion. PMID- 9725775 TI - Expression of growth hormone receptor in the bovine mammary gland during prenatal development. AB - Growth hormone (GH) is known to play a key role in postnatal growth and differentiation. The role of GH and its receptor (GHR) in prenatal development, however, is still controversial. Using reverse transcription polymerase chain reaction (RT-PCR), in situ hybridization (ISH) and immunohistochemistry we demonstrated the presence of GHR mRNA and protein in bovine mammary glands during fetal development. RT-PCR revealed GHR transcripts in fetal mammary glands from the third to the ninth month of pregnancy. By non-radioactive ISH, GHR mRNA was localized in the glandular epithelium, the surrounding mesenchymal cells, endothelial cells of vessels and in the stratum basale of the epidermis of fetal mammary glands. From the sixth month of fetal life onwards, GHR transcripts were also found in the cytoplasm of adipocytes. Immunohistochemical studies using the monoclonal antibody mAb 263 revealed the same distribution pattern as the mRNA. Our results imply that the growth hormone receptor is distinctly expressed in the immature mammary gland, suggesting that GH is involved in growth and differentiation of the fetal mammary gland. PMID- 9725776 TI - Low serum aluminum values in dialysis patients with increased bone aluminum levels. AB - BACKGROUND: Using an HPLC/ETAAS hybrid speciation technique we previously demonstrated iron to have a multifold effect on the binding of aluminum to transferrin by limiting the number of available binding sites and decreasing the affinity of transferrin for aluminum. Theoretically, at a 60% iron-transferrin saturation the aluminum-transferrin fraction in serum should not exceed 30 microg/l. In the present study previous experimental data were confronted with recent clinical observations in patients with either normal iron status or iron overload. PATIENTS AND RESULTS: Serum aluminum levels and iron overload: In 38 dialysis patients with a normal iron status and of whom 63% received Al(OH)3 for phosphate binding 26 (68%) had a serum aluminum level >30 microg/l. On the other hand out of 28 transfusional iron overloaded patients; 68% of them taking Al(OH)3, only 1 subject (4%) had a serum aluminum value in excess of 30 microg/l. Taking patients of both groups receiving Al(OH)3 together a significant (p = 0.001) negative correlation (r = -0.5017) was found between the iron-transferrin saturation and the serum aluminum levels. Iron status and parenteral aluminum loading: Also could a significant (p = 0.001) negative correlation (r = -0.6383) between these parameters be found in an independent group of 44 patients which were acutely intoxicated by the use of aluminum-contaminated dialysis fluids. Since in this population aluminum loading occurred parenterally and not via the gastrointestinal tract, a direct effect of iron on the transferrin binding of aluminum rather than on the element's gastrointestinal absorption must have been responsible for the inverse relationship. Bone aluminum and iron overload: Out of 22 patients with a normal iron status (mean + SD serum ferritin: 216 +/- 245 microg/l; iron-transferrin saturation 20.4 +/- 9.6%), all of them having aluminum overload (bone aluminum level >15 microg/g and/or positive Aluminon staining) none of them presented with a serum aluminum <30 microg/l (mean +/- SD: 82.2 +/- 51.6 microg/l). On the other hand out of 13 iron overloaded patients (serum ferritin >800 microg/l; iron-transferrin saturation 61.4 +/- 17.6%) 10 (77%) presented the proposed criteria of aluminum overload in the presence of a serum aluminum level <30 microg/l. CONCLUSIONS: Our data indicate that in dialysis patients with iron overload (iron-transferrin saturation >60%; serum ferritin >800 microg/l) serum aluminum levels are low (<30 microg/l) despite exposure to aluminum by the intake of Al(OH)3 or the use of aluminum-contaminated dialysis fluids. Low serum aluminum nevertheless may be associated with aluminum overload and even aluminum-related bone disease. An effect of iron on the serum aluminum speciation may at least in part explain our observations. Our findings allow a more accurate interpretation of baseline serum aluminum values. PMID- 9725777 TI - Factors influencing serum aluminum in CAPD patients. AB - The aim of this study was to find out the relationship between body iron stores and serum aluminum levels among 82 stable CAPD patients. The influence of other factors such as time on CAPD and residual renal function was also considered. Thirty-three patients received aluminum hydroxide as a phosphate binder, and they had significantly higher aluminum levels (36.45 microg/l) than the patients who were not taking aluminum preparations (17.2 microg/l, p = 0.001). A statistically significant correlation between serum aluminum levels and residual renal function and time on CAPD was also observed (p <0.05). However, there was no relationship between serum aluminum levels and serum iron, ferritin and transferrin saturation, neither between body iron stores and total excretion of aluminum (p >0.05). In previous reports, low serum iron levels were associated with high serum aluminum concentration among hemodialysis patients. However, this effect was not observed in the CAPD population under study. The highest risk of hyperaluminemia was found in the patients who were taking aluminum hydroxide, had worse residual renal function and had been longer on CAPD. PMID- 9725778 TI - Specialized chronic care for dialysis patients--a five-year study. AB - BACKGROUND: At least 40% of those starting onto renal dialysis at the present time are aged over 65 years old. With old age comes increased comorbidity and decreased functional status. The long term management of older patients is limited by the need for rehabilitation and by placement concerns. We describe a 5 year experience of a pilot program, created in 1991 on the recommendation of the Metropolitan Toronto District Health Council, to rehabilitate and care for elderly and disabled patients on either hemodialysis or peritoneal dialysis. METHODS AND RESULTS: This retrospective, observational study reports on a total of 185 patients admitted over a 5-year period to the Riverdale Chronic Dialysis Unit for chronic care or rehabilitation. The mean age of patients admitted was 67 years (quartiles 61 and 75 years). Eighty-five percent of patients had 2 or more severe comorbidities, while 60% had 3 or more active medical issues. The most commonly used dialysis modality was hemodialysis (80%). Of the 185 patients followed 34% were discharged home, 35% died and 13% were still resident at the time of completion of the study. The most common acute medical problems seen in these patients related to their vascular access and necessitated temporary transfer to an acute nephrology center. A total of 4.7 transfers were recorded for each patient-year of follow up. CONCLUSIONS: This study describes the adaptation of facilities already present in our area, to allow better management and placement of older dialysis patients. Transfer of patients from a high level acute care facility to a chronic care facility makes economic and practical sense and may allow better long term health care planning as well as more stability for the family or care-givers. PMID- 9725779 TI - Dihydropyridine type calcium channel blocker-induced turbid dialysate in patients undergoing peritoneal dialysis. AB - We previously reported that manidipine, a new dihydropyridine type calcium channel blocker, produced chylous peritoneal dialysate being visually indistinguishable from infective peritonitis in 5 patients undergoing continuous ambulatory peritoneal dialysis (CAPD) [Yoshimoto et al. 1993]. To study whether such an adverse drug reaction would also be elicited by other commonly prescribed calcium channel blockers in CAPD patients, we have conducted postal inquiry to 15 collaborating hospitals and an institutional survey in International Medical Center of Japan as to the possible occurrence of calcium channel blocker associated non-infective, turbid peritoneal dialysate in CAPD patients. Our diagnostic criteria for drug-induced turbidity of dialysate as a) it developed within 48 h after the administration of a newly introduced calcium channel blocker to the therapeutic regimen, b) absence of clinical symptoms of peritoneal inflammation (i.e., pyrexia, abdominal pain, nausea or vomiting), c) the fluid containing normal leukocyte counts and being negative for bacterial and fungal culture of the fluid, and d) it disappeared shortly after the withdrawal of the assumed causative agent. Results showed that 19 out of 251 CAPD patients given one of the calcium channel blockers developed non-infective turbid peritoneal dialysis that fulfilled all the above criteria. Four calcium channel blockers were suspected to be associated with the events: benidipine [2 out of 2 (100%) patients given the drug], manidipine [15 out of 36 (42%) patients], nisoldipine [1 out of 11 (9%) patients] and nifedipine [1 out of 159 (0.6%)] in descending order of frequency. None of the patients who received nicardipine, nilvadipine, nitrendipine, barnidipine and diltiazem (25, 7, 2, 1 and 8 patients, respectively) exhibited turbid dialysate. In conclusion, we consider that certain dihydropyridine type calcium channel blockers would cause turbid peritoneal dialysate being similar to that observed in patients developing infective peritonitis. To avoid unnecessary antibiotic therapy the possibility of this adverse reaction should be ruled out whenever a CAPD patient receiving a dihydropyridine type calcium channel blocker develops turbid dialysate. PMID- 9725780 TI - Bone density and skeletal metabolism are altered in idiopathic hypercalciuria. AB - OBJECTIVE: To study bone density in hypercalciuric patients, when classified according to the main metabolic defect. METHODS: We studied 49 patients, aged 19 60 years with calcium stones and idiopathic hypercalcuria. All subjects underwent an evaluation of mineral metabolism and a spinal and femoral DEXA measurement. Then, patients were classified as having Fasting (FH, 31 subjects) or Absorptive (AH, 18 patients) Hypercalciuria according to a standard oral calcium load. RESULTS: Spinal bone density was lower only in FH patients as compared to controls (p <0.001). Bone alkaline phosphatase and urine hydroxyproline were higher with respect to controls only in patients with FH (p <0.005 and p <0.015, respectively). After low calcium diet, hydroxyproline excretion continued to be higher in FH patients (p <0.05). Although in the normal range, serum and urine uric acid were higher in hypercalciuric subjects (p <0.03 and p <0.005, respectively); blood pH was lower in hypercalciuric patients than in controls (p <0.01). In FH patients urine hydroxyproline negatively correlated with spinal and femoral density (p <0.001 and p <0.005, respectively), and blood pH positively correlated with spinal density. CONCLUSIONS: a disordered bone metabolism and bone loss are present only in patients with fasting hypercalciuria. An excessive acid load, possibly of dietary origin, might be involved as a pathogenetic factor. PMID- 9725781 TI - Midodrine is effective and safe therapy for intradialytic hypotension over 8 months of follow-up. AB - Intradialytic hypotension (IDH) is a common and frustrating complication of hemodialysis. Certain end stage renal disease (ESRD) patients recurrently manifest this disabling condition. Both patient-specific factors (autonomic insufficiency, cardiac disease) and dialysis treatment-related factors (ultrafiltration, increased core body temperature) are thought to have significant causative roles. Most therapeutic interventions to date have been either unsuccessful or poorly tolerated. However, recent studies have shown that midodrine, an oral peripheral alpha-1 adrenergic agonist, is an effective and safe therapy for symptomatic IDH in the short-term. We report our experience with the predialysis use of midodrine for IDH in 13 hemodialysis (HD) patients over a 5 to 8 month period. Thirteen patients (8 male, 5 female, mean age 63.9 yrs) with recurrent symptomatic IDH were given midodrine 10 mg orally 30 min before each HD session. Blood pressures (pre-HD BP, lowest intradialytic [ID] BP, post-HD BP) and body weights were tracked for each HD treatment. Values for 10 HD sessions prior to midodrine therapy (Baseline) were compared to values (10 HD sessions each) during the 1st, 5th and 8th month of midodrine therapy. Data were analyzed using ANOVA for repeated measures and paired t-tests, with each patient serving as his/her own control. Patients were monitored for 5 months (n = 13) and 8 months (n = 8), respectively. All lowest intradialytic BPs, post-HD SBPs, and MAPs were significantly improved (p <0.05) on midodrine therapy. This effect was maintained during all periods of follow-up. There was no significant difference in mean albumin, hematocrit, Kt/V, calcium, and sodium between baseline and all periods of follow-up. Mean ultrafiltration volume per HD session was not significantly different than baseline over the course of study. A subjective improvement in hypotensive symptoms was also noted. Importantly, there were no adverse reactions to midodrine in all periods of follow-up. Midodrine appears to be an effective and safe treatment for HD patients with symptomatic IDH, and remains beneficial when used for an extended period of time. PMID- 9725782 TI - Effects of benazepril on insulin resistance and glucose tolerance in uremia. AB - This study tested whether the angiotensin-converting enzyme inhibitor (ACEI) benazepril can improve the insulin resistance and glucose tolerance in uremia. Fifteen uremic hypertensive patients were treated with benazepril in a dose of 10 20 mg per day for ten weeks, and ten healthy subjects, matched in age, sex ratio and body mass index (BMI), served as the control group. Before and after the treatment, an oral 75 g glucose tolerance test (OGTT) and insulin release test (IRT) were performed in two groups above, and the blood glucose and serum insulin concentrations at 0, 60, 120 and 180 minutes after glucose load were examined, and the insulin glycoregulatory activity, including insulin sensitivity index (ISI), glucose uptake rate (M), total areas under the glucose and insulin curves during OGTTs (AUCG AUCINS), was calculated. The changes of serum potassium and renal function before and after treatment were observed. It showed that (1) benazepril could reduce blood pressure significantly (SBP decreased from 174.8 +/ 12.0 mmHg to 151.5 +/- 9.0 mmHg, p <0.001; DBP decreased from 108.0 +/- 8.2 mmHg to 95.3 +/- 9.0 mmHg, p <0.001). The total response rate was 86.7%. (2) After treatment with benazepril for ten weeks, the blood glucose and serum insulin concentrations after glucose load and AUCG, AUCINS values in the uremic patients were significantly lower than before treatment, but were still significantly higher than in the controls. The values of ISI and M in the uremic patients after treatment were much higher than before treatment, but were still significantly lower than in the control subjects. (3) The differences of serum potassium and creatinine levels before and after treatment were not significant. These findings indicate that benazepril can not only reduce blood pressure effectively and safely, but also partly improve insulin resistance, hyperinsulinemia and glucose intolerance in uremia. PMID- 9725784 TI - Successful treatment of severe azathioprine-induced hepatic veno-occlusive disease in a kidney-transplanted patient with transjugular intrahepatic portosystemic shunt. AB - Azathioprine-induced veno-occlusive disease of the liver mainly described after kidney transplantation is as rare as severe with a high mortality due to acute portal hypertension and liver failure. A kidney-transplanted patient with severe azathioprine-induced veno-occlusive disease of the liver and worsening despite drug discontinuation was treated by emergency transjugular intrahepatic portosystemic shunt. Whereas the veno-occlusive disease was controlled, the patient developed severe intractable portosystemic encephalopathy successfully treated by a stent reducer maintaining a certain degree of portal diversion. Twelve months after transjugular intrahepatic portosystemic shunt, liver function was normalized and the stent was thrombosed with a subnormal liver histology. Thirty-six months after transjugular intrahepatic portosystemic shunt the patient is alive with normal liver function tests and kidney graft function. Transjugular intrahepatic portosystemic shunt for treatment of severe veno-occlusive disease of the liver is an alternative to tide the patient over until recovery of liver function. PMID- 9725783 TI - Clinical characteristics of polycystic kidney disease with end-stage renal disease. The Kanazawa Renal Disease Study Group. AB - AIMS: Polycystic kidney disease (PKD) is one of important causes of end-stage renal disease (ESRD). However, there have been few detailed reports on PKD patients with ESRD. The present study was designed to clarify the clinical characteristics of PKD patients with ESRD. SUBJECTS AND METHODS: We collected data from 22 renal divisions in our region, where 63 of 1246 patients with ESRD (male/female: 31/32) were proven to suffer from PKD (5.06%). RESULTS: The average age at the induction of renal replacement therapy was 52.4 +/- 10.0 years. Of these patients, 32 (50.8%) had some family members with apparent PKD. Three (4.8%) and 4 (6.3%) had a history of subarachnoidal hemorrhage and intracerebral hemorrhage, respectively. One (1.6%) suffered from tuberous sclerosis. The prevalence of hypertension treated with antihypertensives, anemia treated with rHuEPO, hepatic cyst, pancreatic cyst, intracranial aneurysm and colonic diverticulum were 66.7%, 58.7%, 85.7%, 16.0%, 33.3% and 50.0%, respectively. CONCLUSION: There was no marked difference in general characteristics or history between the present subjects and those described in previous reports. However, the prevalence of complications seemed to be higher than previously estimated. PMID- 9725785 TI - Nocardiosis in a recently transplanted renal patient. AB - Opportunistic infections are common after renal transplantation because of the use of immunosuppression. Nocardiosis is a rare but important cause of morbidity and mortality among renal transplant recipients. Depending upon the transplant center, the estimated incidence of nocardiosis among renal transplant recipients varies widely from 0 to 20%. We report the first case of nocardiosis in a recently transplanted renal patient maintained on tacrolimus, prednisone, and mycophenolate mofetil. It is likely that the nocardial infection in our patient was related to the development of her diabetes mellitus, and to her early episode of rejection and treatment which included high-dose steroids, and the addition of mycophenolate mofetil. Our case illustrates the importance of maintaining a heightened awareness so that nocardiosis may be diagnosed early and treated successfully. PMID- 9725787 TI - Use of a phosphorus-enriched hemodialysate to prevent hypophosphatemia in a patient with renal failure-related pericarditis. AB - We describe a patient who suffered from renal failure-associated pericarditis and underwent daily 3.5-hour hemodialysis treatments for 17 days. The initially elevated serum phosphorus level gradually fell to below normal on days 11 and 12 as a result of the intensive dialytic therapy. Phosphorus was added to the "base concentrate" of a dual-concentrate, bicarbonate-based dialysate delivery system on days 13 to 17. Because of this phosphorus-enrichment, we were able to maintain the patient's serum phosphorus levels within normal limits in spite of continued daily dialysis treatments. PMID- 9725786 TI - Nocardial brain abscess in a renal transplant recipient successfully treated with triple antimicrobials. AB - Nocardia is a serious opportunistic infection in renal transplant recipients and nocardial brain abscess in these patients has a high mortality. In addition to antimicrobial therapy, treatment usually involves craniotomy and excision of the abscess. We describe a renal transplant recipient maintained on cyclosporine and prednisone developing Nocardia Asteroides brain abscess. After stereotactic aspiration of the abscess, successful treatment was achieved by triple therapy with trimethoprim sulfamethoxazole (TMP/SMX), ceftriaxone and amikacin. The allograft function remained stable. Long-term prophylaxis with TMP/SMX is necessary to prevent the relapse of nocardia. PMID- 9725788 TI - Baroreceptor sensitivity in kidney transplant recipients. PMID- 9725789 TI - Some cephalosporins inhibit serum aminotransferase activities in uremic patients. PMID- 9725790 TI - The role of combination effects on the etiology of malignant nasal tumours in the wood-working industry. AB - Adenocarcinomas of the nose are regarded as an occupational disease in Germany and other European countries, and workers exposed to oak or beech wood-dust in the course of their work receive compensation if they incur adenocarcinoma of the nose. This has prompted a joint research project to record the functional and morphological changes of the nasal mucosa and/or paranasal sinus of 149 exposed subjects and 33 controls in accordance with a defined occupational exposure. To ensure the quantitative and qualitative reliability of the exposure data, 1,349 measurements at the company workplace were taken and analyses of 614 wood samples performed; parallel to this, the genotoxic effects of the most important substances used in the wood-working industry were tested. Apart from this, latency periods and morbidity rates in Germany were investigated. Partial findings of this research projects have been evaluated by the International Agency for Research on Cancer (IARC). According to these evaluations and the findings presented here, the following points can be made: i) Morphological changes in the nasal mucosa after exposure to wood-dust resulted in an increase in cylindrocellular hyperplasias and, in functional terms, a tendency towards improved nasal clearance was observed. Chromium and formaldehyde, on the other hand, tended to give rise to an increase in the number of squamous metaplasias. This might explain the preference for the histological types of adenocarcinoma among subjects exposed to wood-dust. ii) In tissue samples more dysplasias were found among those exposed to oak and beech wood-dust. Subjects exposed to wood preserving agents had dysplasias only if they were simultaneously exposed to oak and beech wood-dust. The latter effect did not quite reach the level of significance (p = 0.07) on account of the low numbers of cases. iii) The investigation of genotoxic effects showed that oak and beech contain genotoxic substances that can be dissolved by means of ethanol and cyclohexane; they also showed that 3 out of 8 wood preservatives, 5 out of 16 stains, and 2 out of 11 paints from the wood-working industry are genotoxic too. Apart from this, lindane and PCP have proved to be genotoxic in the nasal cells of rats and human beings. Analysis of 614 wood samples from wood-preserving agents showed that almost 73% contained agents of this type, even in woods described by the companies as being guaranteed free of wood preservatives. iv) According to an analysis of 147 cases accepted since 1985 as a pensionable occupational illness by the Holz Berufsgenossenschaft (an industrial compensation society for employees in the wood-working industry), the disease was much more apparent in small companies where there is multi-factorial exposure, than in large companies where the exposure factor tends to have a single component. This points to the combined effects of hardwood dusts and chemicals as being the cause. v) According to published findings, the incidence of the disease in England seems to be on the decline. In Germany, increasing latency periods also point to a decline in the number of cases, although both countries have only very recently introduced effective prevention measures against exposure to wood-dust. This also leads to the assumption that wood-dusts cannot be the only cause of this type of cancer. vi) These findings tally with the evaluation by the IARC confirming the special part that hardwood dusts play in the development of nasal cancer. The findings presented here also indicate combined effects as being the cause of this type of cancer. This hypothesis cannot be confirmed until the conclusion of long-term animal experiments, currently being conducted, to test how the effects of chemicals such as lindane, PCP, and chromate compare with use of oak wood-dust. PMID- 9725791 TI - Common and emerging infectious causes of hematological malignancies in the young. AB - Comparative epidemiological studies have for a long time suggested a link (or links) between infectious agents and hematological malignancies in the young. Identification of Epstein-Barr virus (EBV) as the major cause of specific subtypes of Burkitt's lymphoma and Hodgkin's disease 20 and 10 years ago, respectively, and the recent involvement of human T-cell leukemia virus in non Hodgkin's lymphomas of the T-cell lineage in young adults in Jamaica have given further credit to early presumptions that these diseases have an infectious etiology. The spectrum of possibly involved viruses: old, EBV, and new, herpesviruses 6, 7 and 8, and unknown retroviruses - as well as the list of partially or totally unresolved disease entities: Hodgkin's disease in adolescents, non-Hodgkin's lymphomas in the immunocompromised, and acute lymphocytic leukemia - is rapidly expanding. Both direct and indirect transforming effects of the above-mentioned viruses are being rapidly disclosed. However, the complex interaction between the different viruses and other causes of hematological malignancies in the young guarantees that many things remain to be discovered also in the future. PMID- 9725793 TI - The Danish Integrated Antimicrobial Resistance Monitoring and Research Programme (DANMAP). PMID- 9725792 TI - Molecular detection and quantitation of the chemokine RANTES mRNA in neurological brain. AB - Accumulation of T-cells in the brains of patients with neurological disorders prompted a molecular analysis of brain tissue for expression of the chemokine RANTES, which is known to be a T-cell activator and chemoattractant. A fast, sensitive and reproducible technique was developed, based on the polymerase chain reaction and nonradioactive detection. The method could detect and quantitate RANTES in small amounts of brain tissue from all patients with multiple sclerosis, and in some patients with other neural or inflammatory diseases. The data indicate constitutive expression of RANTES in brain from some neurological disorders where its downregulation can have therapeutic benefits. PMID- 9725794 TI - Surveillance of antimicrobial resistance in bacteria isolated from food animals to antimicrobial growth promoters and related therapeutic agents in Denmark. AB - This study was conducted to describe the occurrence of acquired resistance to antimicrobials used for growth promotion among bacteria isolated from swine, cattle and poultry in Denmark. Resistance to structurally related therapeutic agents was also examined. Three categories of bacteria were tested: 1) indicator bacteria (Escherichia coli, Enterococcus faecalis, Enterococcus faecium), 2) zoonotic bacteria (Campylobacter, Salmonella, Yersinia enterocolitica), and 3) animal pathogens (E. coli, Staphylococcus aureus, coagulase-negative staphylococci (CNS), Staphylococcus hyicus, Actinobacillus pleuropneumoniae). All antimicrobials used as growth promoters in Denmark and some structurally related therapeutic agents (in brackets) were included: Avilamycin, avoparcin (vancomycin), bacitracin, carbadox, flavomycin, monensin, olaquindox, salinomycin, spiramycin (erythromycin, lincomycin), tylosin (erythromycin, lincomycin), and virginiamycin (pristinamycin). Bacterial species intrinsically resistant to an antimicrobial were not tested towards that antimicrobial. Breakpoints for growth promoters were established by population distribution of the bacteria tested. A total of 2,372 bacterial isolates collected during October 1995 to September 1996 were included in the study. Acquired resistance to all currently used growth promoting antimicrobials was found. A frequent occurrence of resistance were observed to avilamycin, avoparcin, bacitracin, flavomycin, spiramycin, tylosin and virginiamycin, whereas resistance to carbadox, monensin, olaquindox and salinomycin was less frequent. The occurrence of resistance varied by animal origin and bacterial species. The highest levels of resistance was observed among enterococci, whereas less resistance was observed among zoonotic bacteria and bacteria pathogenic to animals. The association between the occurrence of resistance and the consumption of the antimicrobial is discussed. The results show the present level of resistance to growth promoters in bacteria from food animals in Denmark. They will form the baseline for comparison with future prospective studies, thereby enabling the determination of trends over time. PMID- 9725795 TI - Characteristics of four cowpox virus isolates from Norway and Sweden. AB - We report the first isolation of cowpox virus from a domestic cat in Norway, and the first confirmed isolation of cowpox virus from a human case in Norway. These two Norwegian cowpox virus isolates, as well as two Swedish human isolates, were partially characterized and compared with each other and with cowpox virus Brighton and vaccinia virus strain Western Reserve. Restriction enzyme analysis of the genomes revealed differences between all six viruses examined, but suggested that the two Norwegian isolates are closely related, as are the two Swedish isolates. Restriction endonuclease digestion of genomic DNA demonstrated that one of the Swedish isolates and the two Norwegian isolates have larger genomes than vaccinia virus strain Western Reserve, but smaller than cowpox Brighton. All four Scandinavian isolates lacked a 72 base-pair region within the A-type inclusion body protein gene which is present in the prototype cowpox virus Brighton. PMID- 9725796 TI - Comparisons of DNA-mediated immunization procedures directed against surface glycoproteins of human immunodeficiency virus type-1 and hepatitis B virus. AB - DNA vaccination methods were compared to examine the in vivo expression of HIV-1 gp160 and beta-galactosidase, and the resulting immune response. Beta galactosidase plasmid showed expression rates of 2-5% of muscle fibers with or without pretreatments using bupivacaine or cardiotoxin facilitators 1 or 5 days earlier, respectively. In contrast, HIV gp160 expression was lower in untreated or bupivacaine-treated muscles, but was improved by pretreatment with cardiotoxin. Equal expression of beta-galactosidase and HIV gp160 was obtained using gene gun delivery to the epidermis. Unlike the i.m. in situ expression of gp160, the anti-HIV antibody response did not improve after muscle pretreatments but depended on the vaccination intervals. Gene gun delivery of pMN160 also resulted in a slow and low titered antibody response. In contrast, a single i.m. injection of plasmid encoding another viral envelope, HBsAg, resulted in earlier seroconversion to high titers without the need for pretreatments or boostings. Intradermal inoculation by gene gun using 100-fold less DNA resulted in the same anti-HBsAg antibody profile only after boostings. In contrast to the differences in antibody responses, a specific CTL response was obtained in all cases. Bupivacaine-treated muscles showed an extreme degree of edema with disruption of connective tissue (endo- and mesomysium) and was not well tolerated (4 of 19 mice died). Cardiotoxin created muscle necrosis and occasional (2 of 20 mice) development of fibrotic muscles. It is concluded that in vivo expression cannot be properly predicted using reporter gene experiments and that the resulting immune response does not follow directly with the expression rate. It is suggested that the antibody response may depend primarily on the nature of the antigen expressed rather than the DNA vaccination method. It is proposed that gene gun or i.m. injection be used without pretreatment in the case of DNA vaccination with plasmid encoding HIV MN gp160. PMID- 9725797 TI - Encephalocele with an area mimicking giant cell fibroblastoma. Case report. AB - We report a unique case of encephalocele with an area mimicking giant cell fibroblastoma. A 4-month-old boy had been born with a parietal midline mass of the head, measuring 3x3x1.5 cm. Microscopically, it consisted of fibrous connective tissue and brain tissue. The fibrous tissue revealed spindle cells and multinucleated giant cells lining sinusoid-like spaces. This area was reminiscent of giant cell fibroblastoma. These cells were immunopositive for vimentin and negative for CD34. The giant cell fibroblastoma-like lesion in our case seems to be non-neoplastic and hamartomatous, and we think that the immunohistochemistry using CD34 is useful for differentiating the two lesions. PMID- 9725798 TI - Lung morphometry by unbiased methods in emphysema: bronchial and blood vessel volume, alveolar surface area and capillary length. AB - We have estimated lung volume, bronchial volume, vessel volume, alveolar surface area and capillary length in patients who died of lung failure due to emphysema and chronic obstructive pulmonary disease (COPD) and in patients with no clinical signs of respiratory disease. Unbiased morphometric methods were applied to the right lung. The patients with emphysema had equal lung volumes and bronchial and vessel volumes compared to the control group. The alveolar surface area and surface density were significantly decreased to about 67%, of control values. The capillary length and length density were significantly decreased to about 68% of control values. The proportional decreases in alveolar surface area and capillary length suggest that compensatory capillary proliferation has not occurred. These unbiased morphometric studies of emphysema have yielded results in end-stage emphysema that are comparable to those previously reported using biased methods. However, the unbiased methods may provide new insights when applied to earlier stages of the disease. PMID- 9725799 TI - Overexpression of ras is an independent prognostic factor in colorectal adenocarcinoma. AB - The expression of ras was investigated by using immunohistochemistry in 245 primary colorectal adenocarcinomas and 49 corresponding metastases in the lymph nodes. One hundred and forty-four (59%) of the primary tumours presented as ras positive and 37 (76%) were positive in metastases. The ras expression was positively related to cell proliferation (p=0.01) and significantly increased in tumours with aneuploidy (68%) compared to tumours with diploidy (51%) and tetraploidy (53%, p=0.01). The frequency of ras expression was increased from Dukes' stage A to stages B-D (41% vs 62%, p=0.01). ras expression was compared in 40 paired primary tumours and their corresponding metastases, and the difference in expression did not reach statistical significance (73% vs 83%, p=0.32). In survival analyses, ras overexpression predicted a poor prognosis independent of Dukes' stage, DNA ploidy and S-phase fraction (p=0.049). We did not find any significant relationship between ras expression and patients' sex, age, tumour location, growth pattern, differentiation, p53 expression or heat shock protein. The results indicate that the alteration of ras expression may be involved in the instability of DNA and cellular overproliferation, but not in the progression to advanced stage and the development of metastases. The expression of ras was an important biological marker for evaluating the prognosis in patients with colorectal adenocarcinoma. PMID- 9725800 TI - Managing cough as a defense mechanism and as a symptom. A consensus panel report of the American College of Chest Physicians. PMID- 9725801 TI - Overexpression of IGF-I in skeletal muscle of transgenic mice does not prevent unloading-induced atrophy. AB - This study examined the association between local insulin-like growth factor I (IGF-I) overexpression and atrophy in skeletal muscle. We hypothesized that endogenous skeletal muscle IGF-I mRNA expression would decrease with hindlimb unloading (HU) in mice, and that transgenic mice overexpressing human IGF-I (hIGF I) specifically in skeletal muscle would exhibit less atrophy after HU. Male transgenic mice and nontransgenic mice from the parent strain (FVB) were divided into four groups (n = 10/group): 1) transgenic, weight-bearing (IGF-I/WB); 2) transgenic, hindlimb unloaded (IGF-I/HU); 3) nontransgenic, weight-bearing (FVB/WB); and 4) nontransgenic, hindlimb unloaded (FVB/HU). HU groups were hindlimb unloaded for 14 days. Body mass was reduced (P < 0.05) after HU in both IGF-I (-9%) and FVB mice (-13%). Contrary to our hypothesis, we found that the relative abundance of mRNA for the endogenous rodent IGF-I (rIGF-I) was unaltered by HU in the gastrocnemius (GAST) muscle of wild-type FVB mice. High-level expression of hIGF-I peptide and mRNA was confirmed in the GAST and tibialis anterior (TA) muscles of the transgenic mice. Nevertheless, masses of the GAST and TA muscles were reduced (P < 0.05) in both FVB/HU and IGF-I/HU groups compared with FVB/WB and IGF-I/WB groups, respectively, and the percent atrophy in mass of these muscles did not differ between FVB and IGF-I mice. Therefore, skeletal muscle atrophy may not be associated with a reduction of endogenous rIGF I mRNA level in 14-day HU mice. We conclude that high local expression of hIGF-I mRNA and peptide in skeletal muscle alone cannot attenuate unloading-induced atrophy of fast-twitch muscle in mice. PMID- 9725802 TI - Blockage of gonadotropin-induced first ovulation caused by thyroidectomy and its possible mechanisms in rats. AB - To determine the role of the thyroid gland on the ovarian functions during the initiation process of puberty, we examined the effects and its mechanisms of hypothyroidism on the first ovulation induced by equine chorionic gonadotropin (eCG) in immature female rats. Animals were thyroidectomized on day 22 and were injected with 5 IU of eCG on day 26 to induce the first ovulation on day 29. The number of antral follicles that secrete inhibin and the ovarian weight were significantly increased in thyroidectomized rats (Tx rats) 48 h after eCG treatment compared with those in non-Tx rats. However, thyroidectomy (Tx) significantly suppressed the rates of ovulating animals on day 29. The blockage of ovulation in Tx rats was recovered by administration of human chorionic gonadotropin or luteinizing hormone (LH)-releasing hormone (LHRH) on day 28. Inhibition of serum LH (not follicle-stimulating hormone) levels induced by Tx was almost restored to control levels by injection of LHRH. A significant increment in prolactin levels in Tx rats was also observed on day 28. The present data indicate that Tx before puberty in female rats causes the blockage of the first ovulation and that the inhibitory effects on ovulation are mainly due to the reduction in the preovulatory LH surge, which is partially mediated through an inhibition of LHRH action on the secretion of LH. PMID- 9725804 TI - Acute effects of triiodothyronine on glucose and fatty acid metabolism during reperfusion of ischemic rat hearts. AB - Clinical studies have demonstrated improved myocardial recovery after severe ischemia in response to acute triiodothyronine (T3) treatment. We determined whether T3 improves the recovery of ischemic hearts by improving energy substrate metabolism. Isolated working rat hearts were perfused with 5.5 mM glucose and 1.2 mM palmitate and were subjected to 30 min of no-flow ischemia. Glycolysis, glucose oxidation, and palmitate oxidation were measured during aerobic reperfusion by adding [5-3H]glucose, [U-14C]glucose, or [9,10-3H]palmitate to the perfusate, respectively. During reperfusion, cardiac work in untreated hearts recovered to a lesser extent than myocardial O2 consumption (MVO2), resulting in a decreased recovery of cardiac efficiency, which recovered to only 25% of preischemic values. Treatment of hearts with T3 (10 nM) before ischemia increased glucose oxidation during reperfusion, which was associated with a significant increase in pyruvate dehydrogenase (PDH) activity, the rate-limiting enzyme for glucose oxidation. In contrast, T3 had no effect on MVO2, glycolysis, or palmitate oxidation. This resulted in a significant decrease in H+ production from glycolysis uncoupled from glucose oxidation (2.7 +/- 0.3 and 1.9 +/- 0.3 micromol . g dry wt-1 . min-1 in control and T3-treated hearts, respectively, P < 0.05), as well as a 3.2-fold improvement in cardiac work and a 2.3-fold increase in cardiac efficiency compared with untreated postischemic hearts (P < 0.05). These data suggest that T3 can exert acute effects that improve the coupling of glycolysis to glucose oxidation, thereby decreasing H+ production and increasing cardiac efficiency as well as contractile function during reperfusion of the postischemic heart. PMID- 9725805 TI - Whole body fat oxidation is related to in situ adipose tissue lipolytic response to isoproterenol in males. AB - A high 24-h respiratory quotient (RQ), i.e., low fat oxidation, predicts weight gain. To determine whether impaired fat mobilization (lipolysis) may contribute to weight gain, we studied the relation between lipolytic response to nonselective beta-adrenergic stimulation and RQ measured in a respiratory chamber in 21 males (11 Caucasians, 10 Pima Indians; age 32 +/- 5 yr, weight 93 +/- 24 kg, body fat 30 +/- 8%; means +/- SD) and 23 females (10 Caucasians, 13 Pima Indians; age 32 +/- 9 yr, weight 95 +/- 26 kg, body fat 44 +/- 8%). Lipolytic response was assessed as the relative increase in dialysate glycerol concentration (% above baseline) when isoproterenol (1 micromol/l) was added to the perfusate of a microdialysis probe inserted in the abdominal subcutaneous adipose tissue. In males, but not in females, basal RQ measured during sleep from 0500 to 0630 and adjusted for waist circumference was negatively correlated to lipolytic response (r = -0.66, P = 0.001). The results suggest that in males, impaired beta-adrenergic-mediated lipolysis may contribute to low rates of fat oxidation, a condition known to predispose to weight gain. PMID- 9725803 TI - Bidirectional regulation of uncoupling protein-3 and GLUT-4 mRNA in skeletal muscle by cold. AB - To elucidate the possible role of the mitochondrial uncoupling protein (UCP)-3 in skeletal muscle as a regulator of adaptive thermogenesis and energy balance, we examined the modulation by cold exposure (5 degrees C) of UCP-3 and glucose transporter isoform GLUT-4 mRNAs in male Sprague-Dawley rats. In skeletal muscle, UCP-3 and GLUT-4 mRNAs increased two- to threefold between 6 and 24 h of cold exposure and then decreased to 50% of the control value after 6 days in the cold. In contrast, skeletal muscle UCP-2 mRNA showed a small increase on day 3 and returned to normal after 6 days. The bidirectional regulation of UCP-3 and GLUT-4 mRNAs in skeletal muscle by cold suggests that UCP-3 may be a major mediator of acute adaptive thermogenesis but then is downregulated, along with GLUT-4, in the chronic state to preserve energy. In contrast, cold exposure caused only transient changes of UCP-2 and GLUT-4 mRNA in heart. These data are consistent with the necessity of the heart to continuously expend energy to maintain blood circulation, regardless of environmental conditions. PMID- 9725806 TI - Aging and fasting regulation of leptin and hypothalamic neuropeptide Y gene expression. AB - To investigate the role of aging on the fasting-induced suppression of leptin gene expression and increase in hypothalamic neuropeptide Y (NPY) gene expression, we fasted or fed ad libitum male F-344xBN rats aged 3, 24, and 31 mo for 2 days. We examined leptin mRNA levels in retroperitoneal, inguinal, and epididymal white adipose tissue (WAT); serum leptin levels; and NPY mRNA levels in the hypothalamus. We found that leptin mRNA levels were increased from 3 to 24 mo and leveled off between 24 and 31 mo in both retroperitoneal WAT and inguinal WAT but were unchanged with age in epididymal WAT. Serum leptin levels increased with age, whereas hypothalamic NPY mRNA levels did not change with age. Fasting suppressed leptin gene expression in all three WATs and serum leptin. Moreover, this suppression of serum leptin and of leptin message in retroperitoneal WAT was less in aged rats. Conversely, fasting increased hypothalamic NPY message, again to a lesser extent in aged rats. In both fed (ad libitum) and fasted rats, there was a strong correlation between serum leptin and hypothalamic NPY mRNA levels in the young but not in either of the two aged groups. These data suggest that aged F-344xBN rats are leptin resistant and that the fasting regulation of serum leptin, leptin mRNA, and hypothalamic NPY mRNA is impaired in aged rats. PMID- 9725807 TI - Mutational analysis of the carboxy-terminal phosphorylation site of GLUT-4 in 3T3 L1 adipocytes. AB - The carboxy terminus of GLUT-4 contains a functional internalization motif (Leu 489Leu-490) that helps maintain its intracellular distribution in basal adipocytes. This motif is flanked by the major phosphorylation site in this protein (Ser-488), which may play a role in regulating GLUT-4 trafficking in adipocytes. In the present study, the targeting of GLUT-4 in which Ser-488 has been mutated to alanine (SAG) has been examined in stably transfected 3T3-L1 adipocytes. The trafficking of SAG was not significantly different from that of GLUT-4 in several respects. First, in the absence of insulin, the distribution of SAG was similar to GLUT-4 in that it was largely excluded from the cell surface and was enriched in small intracellular vesicles. Second, SAG exhibited insulin dependent movement to the plasma membrane (4- to 5-fold) comparable to GLUT-4 (4- to 5-fold). Finally, okadaic acid, which has previously been shown to stimulate both GLUT-4 translocation and its phosphorylation at Ser-488, also stimulated the movement of SAG to the cell surface similarly to GLUT-4. Using immunoelectron microscopy, we have shown that GLUT-4 is localized to intracellular vesicles containing the Golgi-derived gamma-adaptin subunit of AP-1 and that this localization is enhanced when Ser-488 is mutated to alanine. We conclude that the carboxy-terminal phosphorylation site in GLUT-4 (Ser-488) may play a role in intracellular sorting at the trans-Golgi network but does not play a major role in the regulated movement of GLUT-4 to the plasma membrane in 3T3-L1 adipocytes. PMID- 9725808 TI - Relation between transamination of branched-chain amino acids and urea synthesis: evidence from human pregnancy. AB - Protein and nitrogen (N) accretion by the mother is a major adaptive response to pregnancy in humans and animals to meet the demands of the growing conceptus. Quantitative changes in whole body N metabolism were examined during normal pregnancy by measuring the rates of leucine N (QN) and carbon (QC) kinetics with the use of [1-13C,15N]leucine. Rate of synthesis of urea was measured by [15N2]urea tracer. Pregnancy-related change in total body water was quantified by H2[18O] dilution, and respiratory calorimetry was performed to quantify substrate oxidation. A significant decrease in the rate of urea synthesis was evident in the 1st trimester (nonpregnant 4.69 +/- 1.14 vs. pregnant 3.44 +/- 1.11 micromol . kg-1 . min-1; means +/- SD, P < 0.05). The lower rate of urea synthesis was sustained through the 2nd and 3rd trimesters. QN was also lower in the 1st trimester during fasting; however, it reached a significant level only in the 3rd trimester (nonpregnant 166 +/- 35 vs. 3rd trimester 135 +/- 16 micromol . kg-1 . h-1; P < 0.05). There was no significant change in QC during pregnancy. A significant decrease in the rate of transamination of leucine was evident in the 3rd trimester both during fasting and in response to nutrient administration (P < 0.05). The rate of deamination of leucine was correlated with the rate of urea synthesis during fasting (r = 0.59, P = 0.001) and during feeding (r = 0.407, P = 0. 01). These data show that pregnancy-related adaptations in maternal N metabolism are evident early in gestation before any significant increase in fetal N accretion. It is speculated that the lower transamination of branched chain amino acids may be due to decreased availability of N acceptors such as alpha-ketoglutarate as a consequence of resistance to insulin action evident in pregnancy. PMID- 9725809 TI - Regulation of glucose production by NEFA and gluconeogenic precursors during chronic glucagon infusion. AB - We previously reported that simulation of the chronic hyperglucagonemia seen during infection was unable to recreate the infection-induced increase in hepatic glucose production. However, chronic hyperglucagonemia was accompanied by a fall in the arterial levels of gluconeogenic precursors as opposed to a rise as is seen during infection. Thus our aim was to determine whether an infusion of gluconeogenic precursors could increase hepatic glucose production in a setting of hyperglucagonemia. Studies were done in 11 conscious chronically catheterized dogs in which sampling (artery and portal and hepatic veins) and infusion catheters (splenic vein) were implanted 17 days before study. Forty-eight hours before infusion of gluconeogenic (GNG) precursors, a sterile fibrinogen clot was placed into the peritoneal cavity. Glucagon was infused over the subsequent 48-h period to simulate the increased glucagon levels (approximately 500 pg/ml) seen during infection. On the day of the experiment, somatostatin was infused peripherally, and basal insulin and simulated glucagon were infused intraportally. After a basal period, a two-step increase in lactate and alanine was initiated (120 min/step; n = 5). Lactate (Delta479 +/- 25 and Delta1, 780 +/- 85 microM; expressed as change from basal in periods I and II, respectively) and alanine (Delta94 +/- 13 and Delta287 +/- 44 microM) levels were increased. Despite increases in net hepatic GNG precursor uptake (Delta0.7 +/- 0.3 and Delta1.1 +/- 0.4 mg glucose . kg-1 . min-1), net hepatic glucose output did not increase. Because nonesterified fatty acid (NEFA) levels fell, in a second series of studies, the fall in NEFA was eliminated. Intralipid and heparin were infused during the two-step substrate infusion to maintain the NEFA levels constant in period I and increase NEFA availability in period II (Delta -29 +/- 29 and Delta689 +/- 186 microM; n = 6). In the presence of similar increases in net hepatic GNG precursor uptake and despite increases in arterial glucose levels (Delta17 +/- 5 and Delta38 +/- 12 mg/dl), net hepatic glucose output increased (Delta0.6 +/- 0.1 and Delta0.7 +/- 0.2 mg . kg-1 . min-1). In summary, a chronic increase in glucagon, when combined with an acute increase in gluconeogenic precursor and maintenance of NEFA supply, increases hepatic glucose output as is seen during infection. PMID- 9725810 TI - The opiate sufentanil alters the inflammatory, endocrine, and metabolic responses to endotoxin in dogs. AB - Sufentanil is a synthetic mu-opioid receptor agonist frequently used in anesthesia and critically ill patients. To evaluate the effects of sufentanil on the inflammatory, neuroendocrine, and metabolic responses to endotoxin, we studied six dogs during saline infusion (control), during sufentanil infusion (1.5 microg . kg-1 . h-1), after endotoxin injection (1.0 microg/kg iv), and during combined endotoxin and sufentanil administration. The rate of appearance of glucose was determined by infusion of [6,6-2H2]glucose. Sufentanil depressed the endotoxin-induced increase in body temperature (36.9 +/- 0.3 vs. 40.6 +/- 0.5 degrees C, P < 0.05). Sufentanil depressed the tumor necrosis factor (TNF) response to endotoxin by approximately 60% (P < 0.01) but increased the interleukin-6 (IL-6) response by approximately 70% (P < 0.01). Sufentanil per se induced a transient neuroendocrine activation. Sufentanil also increased plasma concentrations of insulin and catecholamines after endotoxin (P < 0.05 vs. endotoxin alone) and increased plasma glucose levels by approximately 36% (from 6.1 +/- 0.1 to 8.3 +/- 0.6 mmol/l, P < 0.05 vs. endotoxin alone). Endotoxin stimulated glucose production transiently by 95% (24.2 +/- 3.2 vs. control 12.4 +/- 1.0 micromol . kg-1 . min-1, P < 0.05). Paradoxically, sufentanil inhibited this endotoxin-induced stimulation of glucose production (P < 0.05 vs. endotoxin alone). In conclusion, sufentanil modulates the response to intravenous endotoxin by dissociating the TNF and IL-6 response, increasing insulin and catecholamine levels, and depressing the increase in glucose production. Therefore, opiates alter inflammatory, endocrine, and metabolic regulation in endotoxemia. PMID- 9725811 TI - Effects of epinephrine on glucose metabolism in contracting rat skeletal muscles. AB - The effects of epinephrine on glucose metabolism during contractile activity and insulin stimulation were investigated in fast-twitch (epitrochlearis) and slow twitch (soleus) muscles from Wistar rats. All muscles were mounted on contraction apparatuses, and some muscles were stimulated electrically for 30 min in vitro. Glucose uptake and glucose phosphorylation were measured with 2-[1, 2-3H(N)]deoxy D-glucose and glucose transport with 3-O-[methyl-3H]methyl-D-glucose. D-[1 14C]mannitol was used to correct for extracellular space. In epitrochlearis, both contraction and insulin increased glucose transport by threefold, and combined they showed an additive effect. Epinephrine (10(-6) M) did not influence glucose transport across the membrane during contractile activity or insulin stimulation. In the absence of epinephrine, similar glucose phosphorylation was obtained during contraction and during insulin stimulation in epitrochlearis ( approximately 12 mmol . kg dry wt-1 . 30 min-1). In the presence of epinephrine, 9.5 +/- 0. 6 mmol . kg dry wt-1 . 30 min-1 glucose was phosphorylated during contraction, whereas only 2.0 +/- 0.3 mmol . kg dry wt-1 . 30 min-1 was phosphorylated during insulin stimulation (P < 0.01), despite a similar glucose 6 phosphate concentration. Comparable results were obtained in soleus. In conclusion, our data suggest that epinephrine inhibits glucose phosphorylation much less during contraction than during insulin stimulation. PMID- 9725812 TI - Impaired phasic insulin and amylin secretion in diabetic rats. AB - We have proposed that a hyperstimulated insulin secretion causing beta-cell degranulation is the basis for the impaired glucose-potentiated insulin secretion in type 2 diabetes ("overworked beta-cell"). To confirm this idea, we previously investigated tolbutamide-infused euglycemic rats. Two novel kinds of beta-cell dysfunction were observed: altered phasic glucose-potentiated insulin secretion with preferential sparing of the first phase and a raised secreted ratio of amylin to insulin. The current study tested these parameters in 90% (intact beta cell insulin stores) and 95% (markedly lowered insulin stores) pancreatectomized (Px) diabetic rats. Rats underwent pancreas perfusion 5-6 wk postsurgery. Controls showed nonchanging insulin secretion during a 20-min perfusion of 16.7 mM glucose + 10 mM arginine. In contrast, both Px groups showed an altered phasic pattern, with the first phase being supernormal (for the beta-cell mass) but the second phase reduced in tandem with the insulin content. Amylin secretion from control and 90% Px rats paralleled the insulin output, so that the amylin-to insulin ratio averaged 0. 12 +/- 0.03% in the controls and 0.16 +/- 0.01% in the 90% Px rats over the two secretory phases. In contrast, the amylin-to-insulin ratio in 95% Px rats equaled that of controls during the first phase (0.12 +/- 0.1%) but was twice normal during the second phase (0.32 +/- 0.4%). These results confirm the validity of the overworked beta-cell schema by showing identical beta cell functional defects in Px rats and tolbutamide-infused normoglycemic rats. PMID- 9725813 TI - Age-related differences in the pancreatic beta-cell response to hyperglycemia after eccentric exercise. AB - Eccentric exercise (ECC) causes muscle damage, insulin resistance, and increased pancreatic beta-cell secretion in young individuals. However, the effects of age on the pancreatic beta-cell response to glucose after ECC are unknown. Hyperglycemic clamps (180 min, 10.0 mM) were performed on eight young (age 22 +/- 1 yr) and eight older (age 66 +/- 2 yr) healthy sedentary males without exercise (CONT) and 48 h after ECC. ECC increased (P < 0.02) muscle soreness ratings and plasma creatine kinase concentrations in both groups. Insulin and C-peptide secretions were similar between young and older subjects during CONT clamps. ECC increased (P < 0.05) first-phase (0-10 min) C-peptide area under the curve in young (4.2 +/- 0.4 vs. 3.7 +/- 0.6 nM . min; ECC vs. CONT, respectively) but not in older subjects (3.2 +/- 0.7 vs. 3.5 +/- 0.7 nM . min; ECC vs. CONT), with significant group differences (P < 0.02). Indeed, ECC repressed (P < 0.05) first phase peak C-peptide concentrations in older subjects (0. 93 +/- 0.16 vs. 1.12 +/ 0.11 nM; ECC vs. CONT). Moreover, first-phase C-peptide-to-insulin molar ratios suggest age-related differences (P < 0.05) in insulin/C-peptide clearance after ECC. Furthermore, the observed C-peptide response after ECC was related to abdominal adiposity [r = -0.62, P < 0.02, and r = -0.66, P < 0. 006, for first and second (10-180 min) phases, respectively]. In conclusion, older individuals did not exhibit the compensatory increase in beta-cell secretion observed among young individuals after ECC. Thus, with increasing age, the pancreatic beta-cell may be less responsive to the physiological stress associated with ECC. PMID- 9725814 TI - Palmitate transport and fatty acid transporters in red and white muscles. AB - We performed studies 1) to investigate the kinetics of palmitate transport into giant sarcolemmal vesicles, 2) to determine whether the transport capacity is greater in red muscles than in white muscles, and 3) to determine whether putative long-chain fatty acid (LCFA) transporters are more abundant in red than in white muscles. For these studies we used giant sarcolemmal vesicles, which contained cytoplasmic fatty acid binding protein (FABPc), an intravesicular fatty acid sink. Intravesicular FABPc concentrations were sufficiently high so as not to limit the uptake of palmitate under conditions of maximal palmitate uptake (i.e., 4.5-fold excess in white and 31.3-fold excess in red muscle vesicles). All of the palmitate taken up was recovered as unesterified palmitate. Palmitate uptake was reduced by phloretin (-50%), sulfo-N-succinimidyl oleate (-43%), anti plasma membrane-bound FABP (FABPpm, -30%), trypsin (-45%), and when incubation temperature was lowered to 0 degrees C (-70%). Palmitate uptake was also reduced by excess oleate (-65%), but not by excess octanoate or by glucose. Kinetic studies showed that maximal transport was 1.8-fold greater in red vesicles than in white vesicles. The Michaelis-Menten constant in both types of vesicles was approximately 6 nM. Fatty acid transport protein mRNA and fatty acid translocase (FAT) mRNA were about fivefold greater in red muscles than in white muscles. FAT/CD36 and FABPpm proteins in red vesicles or in homogenates were greater than in white vesicles or homogenates (P < 0.05). These studies provide the first evidence of a protein-mediated LCFA transport system in skeletal muscle. In this tissue, palmitate transport rates are greater in red than in white muscles because more LCFA transporters are available. PMID- 9725816 TI - Lower activity of oxidative key enzymes and smaller fiber areas in skeletal muscle of postobese women. AB - Muscle fiber morphology and activities of four key enzymes, as well as energy metabolism, were determined in nine normal-weight postobese women and nine matched control subjects. No differences in fiber type composition, but a smaller mean fiber area and area of fiber types I and IIb, were found in postobese compared with control subjects (P < 0.05). The activities of beta-hydroxyacyl-CoA dehydrogenase (HADH) and citrate synthase (CS) were 20% lower in postobese than in control subjects (P < 0.05). However, the activities of lactate dehydrogenase and lipoprotein lipase were not significantly different between postobese and control subjects. Basal metabolic rate and respiratory exchange ratio were also similar, but maximal oxygen uptake (VO2 max) tended to be lower in postobese than in control subjects (P = 0.06). When adjustments were made for differences in VO2 max, HADH and CS were not different between postobese and control subjects. In conclusion, these data suggest that smaller fiber areas and lower enzyme activities, i.e., markers of aerobic capacity of skeletal muscle, but not fiber composition, may be factors predisposing to obesity. PMID- 9725815 TI - Endotoxin-induced migration of monocytes and PECAM-1 phosphorylation are abrogated by PAF receptor antagonists. AB - The trafficking of monocytes across the endothelial lining of the blood vessel increases in response to bacterial infection at sites of inflammation. However, the molecular events involved in the diapedesis of monocytes in response to endotoxin are not completely understood. Our studies revealed that signaling by lipopolysaccharide (LPS) in human umbilical vein endothelial cells (HUVEC) resulted in a threefold increase in the transendothelial migration of monocyte like HL-60 cells and a sevenfold increase in the phosphorylation of platelet endothelial cell adhesion molecule-1 (PECAM-1). The transmigration induced by LPS was inhibited by an antibody to PECAM-1. Both the phosphorylation of PECAM-1 and transendothelial migration of monocytes were inhibited by a platelet-activating factor (PAF) receptor antagonist, indicating the autocrine effect of PAF in these events. Treatment of HUVEC with LPS caused a fourfold increase in PAF receptor mRNA expression that was completely blocked by the PAF receptor antagonist. We conclude that PAF, generated by HUVEC in response to LPS or gram-negative bacterial infection, acts in an autocrine manner, causing PECAM-1 phosphorylation and thus the transendothelial migration of monocytes. PMID- 9725817 TI - Plasma leptin concentrations are only transiently increased in nephrectomized rats. AB - Leptin is an adipocyte-secreted hormone that has effects on appetite and energy expenditure. Several studies have shown that end-stage renal disease results in elevated plasma leptin concentrations and that the kidney is responsible for most of leptin elimination in rodents. Leptin metabolism was investigated in rats that underwent unilateral nephrectomy to experimentally limit renal elimination function. Within 4 h of nephrectomy, plasma leptin concentrations increased from 2.9 +/- 0.8 to 5.8 +/- 1.0 & microg/l but thereafter rapidly (<24 h) decreased to prenephrectomy concentrations, despite continued elevated plasma creatinine levels. Sham-operated rats maintained presurgical concentrations of leptin and creatinine throughout the experiment. Kinetic studies of 125I-labeled leptin elimination showed that fractional catabolic rates and half-lives of leptin in circulation were similar at 48 h in nephrectomized and sham-operated rats, suggesting that production of leptin was unchanged after nephrectomy. Excretion of 125I derived from leptin in urine of nephrectomized rats was similar to that of sham-operated rats, and residual radioactivity was increased in the remaining kidneys excised from nephrectomized rats. These results demonstrate that 1) leptin concentrations are quickly restored to presurgical levels in nephrectomized rats, and 2) it is leptin elimination, not leptin production, that compensates to maintain leptin concentrations. Rapid metabolic adaptation of remaining renal tissue may explain the restoration of normal leptin elimination in nephrectomized rats. PMID- 9725818 TI - Arginine-induced insulin release is decreased and glucagon increased in parallel with islet NO production. AB - Nitric oxide (NO) produced by islet constitutive NO synthase (cNOS) is a putative modulator of islet hormone secretion. We show here for the first time that the release of insulin induced by L-arginine or L-homoarginine is inhibited and that of glucagon stimulated in parallel with the rate of islet NO production. It was found that L-homoarginine was approximately 25-30% less potent than L-arginine as an insulin secretagogue but equally potent as a glucagon secretagogue. Biochemical determination of islet cNOS activity revealed that the NO production with L-homoarginine as substrate was only approximately 40% of that of L arginine. Selective inhibition of islet cNOS potentiated insulin release during amino acid stimulation. Moreover, inhibition of cNOS suppressed glucagon release, more so with L-arginine than with L-homoarginine as secretagogue, reflecting the relative rates of their NO production. In K+-depolarized islets, inhibition of cNOS enhanced the insulin response to L-arginine by 50% and that to L homoarginine by 23%, largely corresponding to their relative NO production. The intracellular NO donor hydroxylamine dose dependently inhibited insulin but increased glucagon secretion in K+-depolarized and amino acid-stimulated islets. We conclude that both amino acids have a dual action on insulin release, since their stimulatory effects are reduced in parallel with the rates of their NO production. Furthermore, the greater NO production induced by L-arginine relative to L-homoarginine corresponds to NO-mediated increases in glucagon release. These NO effects are mainly exerted independently of membrane depolarization events. PMID- 9725819 TI - Leptin expression in adipose tissue from obese humans: depot-specific regulation by insulin and dexamethasone. AB - We investigated the in vitro regulation of leptin expression in adipose tissue from severely obese women and men before and after culture with insulin (7 nM) and/or dexamethasone (25 nM). Leptin mRNA and leptin secretion were two- to threefold higher in subcutaneous vs. omental adipose tissue before culture. Dexamethasone transiently increased leptin mRNA approximately twofold in both depots after 1 day of culture [P < 0.01 vs. basal (no hormone control)], but leptin secretion was only increased in omental adipose tissue (P < 0.005 vs. basal). Insulin did not increase leptin mRNA in either depot but increased leptin secretion approximately 1.5- to 3-fold in subcutaneous tissue throughout 7 days of culture (P < 0.05 vs. basal). The combination of insulin and dexamethasone increased leptin mRNA and leptin secretion approximately two- to threefold in both depots at day 1 (P < 0.005 vs. basal or insulin) and maintained leptin expression throughout 7 days of culture. We conclude that insulin and glucocorticoid have depot-specific effects and function synergistically as long term regulators of leptin expression in omental and subcutaneous adipose tissue from obese subjects. PMID- 9725820 TI - Monocarboxylate transporter expression in mouse brain. AB - Although glucose is the major metabolic fuel needed for normal brain function, monocarboxylic acids, i.e., lactate, pyruvate, and ketone bodies, can also be utilized by the brain as alternative energy substrates. In most mammalian cells, these substrates are transported either into or out of the cell by a family of monocarboxylate transporters (MCTs), first cloned and sequenced in the hamster. We have recently cloned two MCT isoforms (MCT1 and MCT2) from a mouse kidney cDNA library. Northern blot analysis revealed that MCT1 mRNA is ubiquitous and can be detected in most tissues at a relatively constant level. MCT2 expression is more limited, with high levels of expression confined to testes, kidney, stomach, and liver and lower levels in lung, brain, and epididymal fat. Both MCT1 mRNA and MCT2 mRNA are detected in mouse brain using antisense riboprobes and in situ hybridization. MCT1 mRNA is found throughout the cortex, with higher levels of hybridization in hippocampus and cerebellum. MCT2 mRNA was detected in the same areas, but the pattern of expression was more specific. In addition, MCT1 mRNA, but not MCT2, is localized to the choroid plexus, ependyma, microvessels, and white matter structures such as the corpus callosum. These results suggest a differential expression of the two MCTs at the cellular level. PMID- 9725821 TI - Restoration of insulin-like growth factor I action in skeletal muscle of old mice. AB - This study examined the effects of long-term chronic voluntary wheel exercise on the ability of insulin-like growth factor I (IGF-I) to stimulate rates of protein synthesis in the soleus muscle of old C57Bl/6 mice. Factors contributing to any changes in hormone action were analyzed at the level of hormone receptor binding, protein content, and gene expression. Chronic exercise resulted in an increased skeletal muscle mass (10-22%) and a 56% increase in IGF-I-stimulated rates of protein synthesis (P < 0.05). IGF-I receptor mRNA was increased 46%, IGF-I receptor protein was increased 65%, and the binding capacity of the IGF-I high affinity site was increased sixfold (P < 0.05) with chronic wheel exercise. Insulin receptor protein content was decreased 35% (P < 0.05), whereas GLUT-4 content was increased 47% with chronic exercise (P < 0.05). This study demonstrates that old animals retain a plasticity for IGF I receptor and glucose transporter expression that may have valuable physiological consequences. PMID- 9725822 TI - Amylin-mediated inhibition of insulin-stimulated glucose transport in skeletal muscle. AB - We examined the effects of amylin on 3-O-methyl-D-glucose (3-O-MG) transport in perfused rat hindlimb muscle under hyperinsulinemic (350 microU/ml, 2,100 pmol/l) conditions. Amylin at 100 nmol/l concentration inhibited 3-O-MG transport relative to control in all three basic muscle fiber types. Transport decreased in slow-twitch oxidative (from 5.65 +/- 1.13 to 3.46 +/- 0.71 micromol . g-1 . h-1), fast-twitch oxidative (from 6.84 +/- 0.90 to 4.84 +/- 0.76 micromol . g-1 . h-1), and fast-twitch glycolytic (from 1.27 +/- 0.20 to 0.60 +/- 0.05 micromol . g-1 . h-1) muscle. Amylin inhibition of insulin-stimulated glucose transport in skeletal muscle was accompanied by a 433 +/- 72% increase in intracellular glucose 6-phosphate (G-6-P) despite the absence of extracellular glucose. The source of hexose units for the formation and maintenance of G-6-P was likely glycogen. Amylin increased glycogenolysis, increased lactate formation, and decreased glycogen synthase activity. Furthermore, the kinetics of glycogen synthase suggest that this enzyme may control intracellular G-6-P concentration. Despite the large increase in G-6-P, no detectable increase in uridine diphosphate-N-acetylhexosamines occurred, suggesting that the proposed glucosamine pathway may not be involved in transport inhibition. However, decreases in uridine diphosphate hexoses were detected. Therefore, uridine or hexosamine-based metabolites may be involved in amylin action. PMID- 9725823 TI - Gluconeogenesis and the Cori cycle in 12-, 20-, and 40-h-fasted humans. AB - Six subjects were infused with [U-13C]glucose (0.03-0.05 mg . kg-1 . min-1) starting 8-9 h after a meal, and the production of glucose, the recycling of glucose (the Cori cycle), the dilution of glucose by unlabeled carbon into the hepatic lactate-pyruvate pool, and gluconeogenesis were determined in these fasted volunteers by use of mass isotopomer analysis and equations previously described [J. A. Tayek and J. Katz. Am. J. Physiol. 272 (Endocrinol. Metab. 35): E476-E484, 1997]. A primed continuous 11-h infusion was started at 6:00 AM, and the above parameters were calculated after 3 h (for the 12-h fast) and at the end of the infusion (for the 20-h fast). Another group of five subjects was fasted for 40 h, and the above parameters were calculated as before. At 12, 20, and 40 h of fasting, respectively, blood glucose was 93 +/- 2, 83 +/- 2, and 71 +/- 2 (SE) mg/dl; glucose production was 2.3, 1.8, and 1.77 mg . kg-1 . min-1; the recycling of labeled carbon was 8, 15, and 15%, and that of glucose molecules (Cori cycle) was 18, 35, and 36%; the contribution of gluconeogenesis to glucose production was 41, 71, and 92% or 0.96, 1.29, and 1.64 mg . kg-1 . min-1; and the contribution of other sources to glucose production was 1.37, 0.53, and 0.15 mg . kg-1 . min-1. The recycling of glucose is important in prolonged fasting for the maintenance of plasma glucose concentration. We demonstrate here that gluconeogenesis can be easily measured and that it accounts for approximately 90% of glucose production after a 40-h fast. PMID- 9725824 TI - Gender differences in leptin levels during puberty are related to the subcutaneous fat depot and sex steroids. AB - Little is known about the influence of adiposity and hormone release on leptin levels in children and adolescents. We utilized criterion methods to examine the relationships among sex steroids, body composition (4 compartment), abdominal visceral and subcutaneous fat (magnetic resonance imagery), total subcutaneous fat (sum of 9 skinfolds), energy expenditure (doubly labeled water), aerobic fitness, and serum leptin levels in prepubertal and pubertal boys (n = 16; n = 13) and girls (n = 12; n = 15). The sum of skinfolds accounted for more variance in leptin levels of all girls [coefficient of determination (R2) = 0.70, P < 0.001] and all boys (R2 = 0.60, P < 0.001) than the total fat mass (girls, R2 = 0.52, P < 0.001; boys, R2 = 0.23, P < 0.001). Total energy expenditure, corrected for the influence of fat-free mass, correlated inversely with leptin (R2 = 0.18, P = 0.02). Gender differences in leptin disappeared when corrected for sex steroid levels or the combination of adiposity and energy expenditure. In multiple regression, the sum of skinfolds and free testosterone and estrogen levels accounted for 74% of the variance in leptin levels. We conclude that serum leptin levels are positively related to subcutaneous adiposity but negatively related to androgen levels. Energy expenditure may be negatively related to leptin levels by reduction of the adiposity, or a common genetic factor may influence both the activity and serum leptin levels. PMID- 9725825 TI - Genes and chromomeres: A puzzle in three dimensions. PMID- 9725826 TI - Characterization of an allele-nonspecific intragenic suppressor in the yeast plasma membrane H+-ATPase gene (Pma1). AB - We have analyzed the ability of A165V, V169I/D170N, and P536L mutations to suppress pma1 dominant lethal alleles and found that the P536L mutation is able to suppress the dominant lethality of the pma1-R271T, -D378N, -D378E, and -K474R mutant alleles. Genetic and biochemical analyses of site-directed mutants at Pro 536 suggest that this amino acid may not be essential for function but is important for biogenesis of the ATPase. Proteins encoded by dominant lethal pma1 alleles are retained in the endoplasmic reticulum, thus interfering with transport of wild-type Pma1. Immunofluorescence studies of yeast conditionally expressing revertant alleles show that the mutant enzymes are correctly located at the plasma membrane and do not disturb targeting of the wild-type enzyme. We propose that changes in Pro-536 may influence the folding of the protein encoded by a dominant negative allele so that it is no longer recognized and retained as a misfolded protein by the endoplasmic reticulum. PMID- 9725827 TI - The [KIL-d] cytoplasmic genetic element of yeast results in epigenetic regulation of viral M double-stranded RNA gene expression. AB - [KIL-d] is a cytoplasmically inherited genetic trait that causes killer virus infected cells of Saccharomyces cerevisiae to express the normal killer phenotypes in a/alpha cells, but to show variegated defective killer phenotypes in a or alpha type cells. Mating of [KIL-d] haploids results in "healing" of their phenotypic defects, while meiosis of the resulting diploids results in "resetting" of the variegated, but mitotically stable, defects. We show that [KIL d] does not reside on the double-stranded RNA genome of killer virus. Thus, the [KIL-d] effect on viral gene expression is epigenetic in nature. Resetting requires nuclear events of meiosis, since [KIL-d] can be cytoplasmically transmitted during cytoduction without causing defects in killer virus expression. Subsequently, mating of these cytoductants followed by meiosis generates spore clones expressing variegated defective phenotypes. Cytoduction of wild-type cytoplasm into a phenotypically defective [KIL-d] haploid fails to heal, nor does simultaneous or sequential expression of both MAT alleles cause healing. Thus, healing is not triggered by the appearance of heterozygosity at the MAT locus, but rather requires the nuclear fusion events which occur during mating. Therefore, [KIL-d] appears to interact with the nucleus in order to exert its effects on gene expression by the killer virus RNA genome. PMID- 9725828 TI - Identification of a calcineurin-independent pathway required for sodium ion stress response in Saccharomyces cerevisiae. AB - The calcium-dependent protein phosphatase calcineurin plays an essential role in ion homeostasis in yeast. In this study, we identify a parallel ion stress response pathway that is independent of the calcineurin signaling pathway. Cells with null alleles in both STD1 and its homologue, MTH1, manifest numerous phenotypes observed in calcineurin mutants, including sodium, lithium, manganese, and hydroxyl ion sensitivity, as well as alpha factor toxicity. Furthermore, increased gene dosage of STD1 suppresses the ion stress phenotypes in calcineurin mutants and confers halotolerance in wild-type cells. However, Std1p functions in a calcineurin-independent ion stress response pathway, since a std1 mth1 mutant is FK506 sensitive under conditions of ion stress. Mutations in other genes known to regulate gene expression in response to changes in glucose concentration, including SNF3, RGT2, and SNF5, also affect cell growth under ion stress conditions. Gene expression studies indicate that the regulation of HAL1 and PMR2 expression is affected by STD1 gene dosage. Taken together, our data demonstrate that response to ion stress requires the participation of both calcineurin dependent and -independent pathways. PMID- 9725829 TI - Identification of functional connections between calmodulin and the yeast actin cytoskeleton. AB - One of four intragenic complementing groups of temperature-sensitive yeast calmodulin mutations, cmd1A, results in a characteristic functional defect in actin organization. We report here that among the complementing mutations, a representative cmd1A mutation (cmd1-226: F92A) is synthetically lethal with a mutation in MYO2 that encodes a class V unconventional myosin with calmodulin binding domains. Gel overlay assay shows that a mutant calmodulin with the F92A alteration has severely reduced binding affinity to a GST-Myo2p fusion protein. Random replacement and site-directed mutagenesis at position 92 of calmodulin indicate that hydrophobic and aromatic residues are allowed at this position, suggesting an importance of hydrophobic interaction between calmodulin and Myo2p. To analyze other components involved in actin organization through calmodulin, we isolated and characterized mutations that show synthetic lethal interaction with cmd1-226; these "cax" mutants fell into five complementation groups. Interestingly, all the mutations themselves affect actin organization. Unlike cax2, cax3, cax4, and cax5 mutations, cax1 shows allele-specific synthetic lethality with the cmd1A allele. CAX1 is identical to ANP1/GEM3/MCD2, which is involved in protein glycosylation. CAX4 is identical to the ORF YGR036c, and CAX5 is identical to MNN10/SLC2/BED1. We discuss possible roles for Cax proteins in the regulation of the actin cytoskeleton. PMID- 9725831 TI - The Saccharomyces cerevisiae RAD9, RAD17, RAD24 and MEC3 genes are required for tolerating irreparable, ultraviolet-induced DNA damage. AB - In wild-type Saccharomyces cerevisiae, a checkpoint slows the rate of progression of an ongoing S phase in response to exposure to a DNA-alkylating agent. Mutations that eliminate S phase regulation also confer sensitivity to alkylating agents, leading us to suggest that, by regulating the S phase rate, cells are either better able to repair or better able to replicate damaged DNA. In this study, we determine the effects of mutations that impair S phase regulation on the ability of excision repair-defective cells to replicate irreparably UV damaged DNA. We assay survival after UV irradiation, as well as the genetic consequences of replicating a damaged template, namely mutation and sister chromatid exchange induction. We find that RAD9, RAD17, RAD24, and MEC3 are required for UV-induced (although not spontaneous) mutagenesis, and that RAD9 and RAD17 (but not REV3, RAD24, and MEC3) are required for maximal induction of replication-dependent sister chromatid exchange. Therefore, checkpoint genes not only control cell cycle progression in response to damage, but also play a role in accommodating DNA damage during replication. PMID- 9725830 TI - Spe3, which encodes spermidine synthase, is required for full repression through NRE(DIT) in Saccharomyces cerevisiae. AB - We previously identified a transcriptional regulatory element, which we call NRE(DIT), that is required for repression of the sporulation-specific genes, DIT1 and DIT2, during vegetative growth of Saccharomyces cerevisiae. Repression through this element is dependent on the Ssn6-Tup1 corepressor. In this study, we show that SIN4 contributes to NRE(DIT)-mediated repression, suggesting that changes in chromatin structure are, at least in part, responsible for regulation of DIT gene expression. In a screen for additional genes that function in repression of DIT (FRD genes), we recovered alleles of TUP1, SSN6, SIN4, and ROX3 and identified mutations comprising eight complementation groups of FRD genes. Four of these FRD genes appeared to act specifically in NRE(DIT)mediated repression, and four appeared to be general regulators of gene expression. We cloned the gene complementing the frd3-1 phenotype and found that it was identical to SPE3, which encodes spermidine synthase. Mutant spe3 cells not only failed to support complete repression through NRE(DIT) but also had modest defects in repression of some other genes. Addition of spermidine to the medium partially restored repression to spe3 cells, indicating that spermidine may play a role in vivo as a modulator of gene expression. We suggest various mechanisms by which spermidine could act to repress gene expression. PMID- 9725832 TI - Roles of prenyl protein proteases in maturation of Saccharomyces cerevisiae a factor. AB - In eukaryotes small secreted peptides are often proteolytically cleaved from larger precursors. In Saccharomyces cerevisiae multiple proteolytic processing steps are required for production of mature 12-amino-acid a-factor from its 36 amino-acid precursor. This study provides additional genetic data supporting a direct role for Afc1p in cleavage of the carboxyl-terminal tripeptide from the CAAX motif of the prenylated a-factor precursor. In addition, Afc1p had a second role in a-factor processing that was independent of, and in addition to, its role in the carboxyl-terminal processing in vivo. Using ubiquitin-a-factor fusions we confirmed that the pro-region of the a-factor precursor was not required for production of the mature pheromone. However, the pro-region of the a-factor precursor contributed quantitatively to a-factor production. PMID- 9725833 TI - Genetic analysis of the Caenorhabditis elegans MAP kinase gene mpk-1. AB - The Caenorhabditis elegans mpk-1 gene encodes a MAP kinase protein that plays an important role in Ras-mediated induction of vulval cell fates. We show that mutations that eliminate mpk-1 activity result in a highly penetrant, vulvaless phenotype. A double mutant containing a gain-of-function mpk-1 mutation and a gain-of-function mek mutation (MEK phosphorylates and activates MPK-1) exhibits a multivulva phenotype. These results suggest that mpk-1 may transduce most or all of the anchor cell signal. Epistasis analysis suggests that mpk-1 acts downstream of mek-2 (encodes a MEK homolog) and upstream of lin-1 (encodes an Ets transcription factor) in the anchor cell signaling pathway. Finally, mpk-1 may act together with let-60 ras in multiple developmental processes, as mpk-1 mutants exhibit nearly the same range of developmental phenotypes as let-60 ras mutants. PMID- 9725834 TI - Genetic and molecular characterization of the caenorhabditis elegans gene, mel 26, a postmeiotic negative regulator of mei-1, a meiotic-specific spindle component. AB - We have previously described the gene mei-1, which encodes an essential component of the Caenorhabditis elegans meiotic spindle. When ectopically expressed after the completion of meiosis, mei-1 protein disrupts the function of the mitotic cleavage spindles. In this article, we describe the cloning and the further genetic characterization of mel-26, a postmeiotic negative regulator of mei-1. mel-26 was originally identified by a gain-of-function mutation. We have reverted this mutation to a loss-of-function allele, which has recessive phenotypes identical to the dominant defects of its gain-of-function parent. Both the dominant and recessive mutations of mel-26 result in mei-1 protein ectopically localized in mitotic spindles and centrosomes, leading to small and misoriented cleavage spindles. The loss-of-function mutation was used to clone mel-26 by transformation rescue. As suggested by genetic results indicating that mel-26 is required only maternally, mel-26 mRNA was expressed predominantly in the female germline. The gene encodes a protein that includes the BTB motif, which is thought to play a role in protein-protein interactions. PMID- 9725835 TI - Two pleiotropic classes of daf-2 mutation affect larval arrest, adult behavior, reproduction and longevity in Caenorhabditis elegans. AB - The nematode Caenorhabditis elegans responds to overcrowding and scarcity of food by arresting development as a dauer larva, a nonfeeding, long-lived, stress resistant, alternative third-larval stage. Previous work has shown that mutations in the genes daf-2 (encoding a member of the insulin receptor family) and age-1 (encoding a PI 3-kinase) result in constitutive formation of dauer larvae (Daf c), increased adult longevity (Age), and increased intrinsic thermotolerance (Itt). Some daf-2 mutants have additional developmental, behavioral, and reproductive defects. We have characterized in detail 15 temperature-sensitive and 1 nonconditional daf-2 allele to investigate the extent of daf-2 mutant defects and to examine whether specific mutant traits correlate with each other. The greatest longevity seen in daf-2 mutant adults was approximately three times that of wild type. The temperature-sensitive daf-2 mutants fell into two overlapping classes, including eight class 1 mutants, which are Daf-c, Age, and Itt, and exhibit low levels of L1 arrest at 25.5 degrees. Seven class 2 mutants also exhibit the class 1 defects as well as some or all of the following: reduced adult motility, abnormal adult body and gonad morphology, high levels of embryonic and L1 arrest, production of progeny late in life, and reduced brood size. The strengths of the Daf-c, Age, and Itt phenotypes largely correlated with each other but not with the strength of class 2-specific defects. This suggests that the DAF-2 receptor is bifunctional. Examination of the null phenotype revealed a maternally rescued egg, L1 lethal component, and a nonconditional Daf c component. With respect to the Daf-c phenotype, the dauer-defective (Daf-d) mutation daf-12(m20) was epistatic to daf-2 class 1 alleles but not the severe class 2 alleles tested. All daf-2 mutant defects were suppressed by the daf-d mutation daf-16(m26). Our findings suggest a new model for daf-2, age-1, daf-12, and daf-16 interactions. PMID- 9725836 TI - Molecular evolution of the Cecropin multigene family in Drosophila. functional genes vs. pseudogenes. AB - Approximately 4 kb of the Cecropin cluster region have been sequenced in nine lines of Drosophila melanogaster and one line of the sibling species D. simulans, D. mauritiana, and D. sechellia. This region includes three functional genes (CecA1, CecA2, and CecB), which are involved in the insect immune response, and two pseudogenes (CecPsi1 and CecPsi2). The level of silent polymorphism in the three Cec genes is rather high (0.028), and there is no excess of nonsynonymous polymorphism. There is no evidence of gene conversion in the history of these genes. The interspecific comparison has revealed that in the three species of the simulans cluster the CecA2 gene is partially deleted and has therefore lost its function and become a pseudogene; in each of the species, subsequent deletions have accumulated. Divergence estimates indicate that the CecPsi1 and CecPsi2 pseudogenes are highly diverged, both between themselves and relative to the other three Cec genes. However, both CecPsi1 and CecPsi2 have conserved transcriptional signals and splice sites, and they present an open reading frame; also, correctly spliced transcripts have been detected for both CecPsi1 and CecPsi2. The data support that these genes are either active genes with some null alleles or young pseudogenes. PMID- 9725837 TI - Induced chromosomal exchange directs the segregation of recombinant chromatids in mitosis of Drosophila. AB - In meiosis, the segregation of chromosomes at the reductional division is accomplished by first linking homologs together. Genetic exchange generates the bivalents that direct regular chromosome segregation. We show that genetic exchange in mitosis also generates bivalents and that these bivalents direct mitotic chromosome segregation. After FLP-mediated homologous recombination in G2 of the cell cycle, recombinant chromatids consistently segregate away from each other (x segregation). This pattern of segregation also applies to exchange between heterologs. Most, or all, cases of non-x segregation are the result of exchange in G1. Cytological evidence is presented that confirms the existence of the bivalents that direct this pattern of segregation. Our results implicate sister chromatid cohesion in maintenance of the bivalent. The pattern of chromatid segregation can be altered by providing an additional FRT at a more proximal site on one chromosome. We propose that sister chromatid exchange occurs at the more proximal site, allowing the recombinant chromatids to segregate together. This also allowed the recovery of reciprocal translocations following FLP-mediated heterologous recombination. The observation that exchange can generate a bivalent in mitotic divisions provides support for a simple evolutionary relationship between mitosis and meiosis. PMID- 9725838 TI - Analysis in Drosophila melanogaster of the interaction between sex combs reduced and extradenticle activity in the determination of tarsus and arista identity. AB - Sex Combs Reduced (SCR) activity is proposed to be required cell nonautonomously for determination of tarsus identity, and Extradenticle (EXD) activity is required cell autonomously for determination of arista identity. Using the ability of Proboscipedia to inhibit the SCR activity required for determination of tarsus identity, we found that loss-of-EXD activity is epistatic to loss-of SCR activity in tarsus vs. arista determination. This suggests that in the sequence leading to arista determination SCR activity is OFF while EXD activity is ON, and in the sequence leading to tarsus determination SCR activity is ON, which turns EXD activity OFF. Immunolocalization of EXD in early third-instar larval imaginal discs reveals that EXD is localized in the nuclei of antennal imaginal disc cells and localized in the cytoplasm of distal imaginal leg disc cells. We propose that EXD localized to the nucleus suppresses tarsus determination and activates arista determination. We further propose that in the mesodermal adepithelial cells of the leg imaginal discs, SCR is required for the synthesis of a tarsus-inducer that when secreted acts on the ectoderm cells inhibiting nuclear accumulation of EXD, such that tarsus determination is no longer suppressed and arista determination is no longer activated. PMID- 9725839 TI - Van Gogh: a new Drosophila tissue polarity gene. AB - Mutations in the Van Gogh gene result in the altered polarity of adult Drosophila cuticular structures. On the wing, Van Gogh mutations cause an altered polarity pattern that is typical of mutations that inactivate the frizzled signaling/signal transduction pathway. The phenotype however, differs from those seen previously, as the number of wing cells forming more than one hair is intermediate between that seen previously for typical frizzled-like or inturned like mutations. Consistent with Van Gogh being involved in the function of the frizzled signaling/signal transduction pathway, Van Gogh mutations show strong interactions with mutations in frizzled and prickle. Mitotic clones of Van Gogh display domineering cell nonautonomy. In contrast to frizzled clones, Van Gogh clones alter the polarity of cells proximal (and in part anterior and posterior) but not distal to the clone. In further contrast to frizzled clones, Van Gogh clones cause neighboring wild-type hairs to point away from rather than toward the clone. This anti-frizzled type of domineering nonautonomy and the strong genetic interactions seen between frizzled and Van Gogh suggested the possibility that Van Gogh was required for the noncell autonomous function of frizzled. As a test of this possibility we induced frizzled clones in a Van Gogh mutant background and Van Gogh clones in a frizzled mutant background. In both cases the domineering nonautonomy was suppressed consistent with Van Gogh being essential for frizzled signaling. PMID- 9725841 TI - The female-determining gene F of the housefly, Musca domestica, acts maternally to regulate its own zygotic activity. AB - In Musca domestica, the common housefly, female development requires the continuous activity of the sex-determining gene F from early embryogenesis until metamorphosis. To activate F in embryogenesis, two conditions must be met: There must be no male-determining M factor in the zygotic genome, and the egg must be preconditioned by F activity in the maternal germ line. This maternal activity can be suppressed by introducing an M factor into the maternal germ line, which causes all offspring, including those that do not carry M, to develop as males. By transplantation of pole cells (germline progenitor cells) we have constructed such females with a genetically male germ line and, simultaneously, males with a genetically female germ line carrying a constitutive allele of F [F(Dominant) (F(D))]. Crosses between these animals yielded offspring that, despite the presence of M in the maternal germ line, were of female sex, solely due to zygotic F(D) brought in via the sperm. This shows that zygotic F function alone is sufficient to promote female development and that in the wild-type situation, maternal F product serves no other function but to activate the zygotic F gene. PMID- 9725840 TI - A genetic screen for novel components of the notch signaling pathway during Drosophila bristle development. AB - The Notch receptor is the central element in a cell signaling mechanism controlling a broad spectrum of cell fate choices. Genetic modifier screens in Drosophila and subsequent molecular studies have identified several Notch pathway components, but the biochemical nature of signaling is still elusive. Here, we report the results of a genetic modifier screen of the bristle phenotype of a gain-of-function Notch allele, Abruptex16. Abruptex mutations interfere with lateral inhibition/specification events that control the segregation of epidermal and sensory organ precursor lineages, thus inhibiting bristle formation. Mutations that reduce Notch signaling suppress this phenotype. This screen of approximately 50,000 flies led to the identification of a small number of dominant suppressors in seven complementation groups. These include known components in the pathway, Notch, mastermind, Delta, and Hairless, as well as two novel mutations. The first, A122, appears to interact with Notch only during bristle development. The other, M285, displays extensive genetic interactions with the Notch pathway elements and appears, in general, capable of suppressing Notch gain-of-function phenotypes while enhancing Notch loss-of-function phenotypes, suggesting that it plays an important role in Notch signaling. PMID- 9725842 TI - Wolbachia transfer from Drosophila melanogaster into D. simulans: Host effect and cytoplasmic incompatibility relationships. AB - Wolbachia are maternally transmitted endocellular bacteria causing a reproductive incompatibility called cytoplasmic incompatibility (CI) in several arthropod species, including Drosophila. CI results in embryonic mortality in incompatible crosses. The only bacterial strain known to infect Drosophila melanogaster (wDm) was transferred from a D. melanogaster isofemale line into uninfected D. simulans isofemale lines by embryo microinjections. Males from the resulting transinfected lines induce >98% embryonic mortality when crossed with uninfected D. simulans females. In contrast, males from the donor D. melanogaster line induce only 18 32% CI on average when crossed with uninfected D. melanogaster females. Transinfected D. simulans lines do not differ from the D. melanogaster donor line in the Wolbachia load found in the embryo or in the total bacterial load of young males. However, >80% of cysts are infected by Wolbachia in the testes of young transinfected males, whereas only 8% of cysts are infected in young males from the D. melanogaster donor isofemale line. This difference might be caused by physiological differences between hosts, but it might also involve tissue specific control of Wolbachia density by D. melanogaster. The wDm-transinfected D. simulans lines are unidirectionally incompatible with strains infected by the non-CI expressor Wolbachia strains wKi, wMau, or wAu, and they are bidirectionally incompatible with strains infected by the CI-expressor Wolbachia strains wHa or wNo. However, wDm-infected males do not induce CI toward females infected by the CI-expressor strain wRi, which is found in D. simulans continental populations, while wRi-infected males induce partial CI toward wDm infected females. This peculiar asymmetrical pattern could reflect an ongoing divergence between the CI mechanisms of wRi and wDm. It would also confirm other results indicating that the factor responsible for CI induction in males is distinct from the factor responsible for CI rescue in females. PMID- 9725843 TI - Intra- and interspecies variation among Bari-1 elements of the melanogaster species group. AB - We have investigated the distribution of sequences homologous to Bari-1, a Tc1 like transposable element first identified in Drosophila melanogaster, in 87 species of the Drosophila genus. We have also isolated and sequenced Bari-1 homologues from D. simulans, D. mauritiana, and D. sechellia, the species constituting with D. melanogaster the melanogaster complex, and from D. diplacantha and D. erecta, two phylogenetically more distant species of the melanogaster group. Within the melanogaster complex the Bari-1 elements are extremely similar to each other, showing nucleotide identity values of at least 99.3%. In contrast, Bari-1-like elements from D. diplacantha and D. erecta are on average only 70% similar to D. melanogaster Bari-1 and are usually defective due to nucleotide deletions and/or insertions in the ORFs encoding their transposases. In D. erecta the defective copies are all located in the chromocenter and on chromosome 4. Surprisingly, while D. melanogaster Bari-1 elements possess 26-bp inverted terminal repeats, their D. diplacantha and D. erecta homologues possess long inverted terminal repeats similar to the terminal structures observed in the S elements of D. melanogaster and in several other Tc1 like elements of different organisms. This finding, together with the nucleotide and amino acid identity level between D. diplacantha and D. erecta elements and Bari-1 of D. melanogaster, suggests a common evolutionary origin and a rapid diversification of the termini of these Drosophila Tc1-like elements. PMID- 9725844 TI - Interactions among dosage-dependent trans-acting modifiers of gene expression and position-effect variegation in Drosophila. AB - We have investigated the effect of dosage-dependent trans-acting regulators of the white eye color gene in combinations to understand their interaction properties. The consequences of the interactions will aid in an understanding of aneuploid syndromes, position-effect variegation (PEV), quantitative traits, and dosage compensation, all of which are affected by dosage-dependent modifiers. Various combinations modulate two functionally related transcripts, white and scarlet, differently. The overall trend is that multiple modifiers are noncumulative or epistatic to each other. In some combinations, developmental transitions from larvae to pupae to adults act as a switch for whether the effect is positive or negative. With position-effect variegation, similar responses were found as with gene expression. The highly multigenic nature of dosage-sensitive modulation of both gene expression and PEV suggests that dosage effects can be progressively transduced through a series of steps in a hierarchical manner. PMID- 9725845 TI - Increased transmitter release and aberrant synapse morphology in a Drosophila calmodulin mutant. AB - The ubiquitous calcium-binding protein calmodulin (CaM) has been implicated in the development and function of the nervous system in a variety of eukaryotic organisms. We have generated mutations in the single Drosophila Calmodulin (Cam) gene and examined the effects of these mutations on behavior, synaptic transmission at the larval neuromuscular junction, and structure of the larval motor nerve terminal. Flies hemizygous for Cam3c1, a mutation in the first Ca2+ binding site, exhibit behavioral, neurophysiological, and neuroanatomical abnormalities. In particular, adults exhibit defects in locomotion, coordination, and flight. Larvae exhibit increased neurotransmitter release from the motor nerve terminal at low [Ca2+] in the presence of the K+ channel-blocking drug quinidine. In addition, synaptic bouton structure at motor nerve terminals is altered. These effects are distinct from those produced by altering the activity of the CaM target enzymes CaM-activated kinase II (CaMKII) and CaM-activated adenylyl cyclase (CaMAC). Furthermore, previous in vitro studies of mutant Cam3c1 demonstrated that although its Ca2+ affinity is decreased, Cam3c1 protein can activate CaMKII, CaMAC, and CaM-activated phosphatase calcineurin in a manner similar to wild-type CaM. Thus, the Cam3c1 mutation might affect Ca2+ buffering or interfere with the activation or inhibition of a CaM target distinct from CaMKII or CaMAC. PMID- 9725846 TI - A composite genetic map of the parasitoid wasp Trichogramma brassicae based on RAPD markers. AB - Three linkage maps of the genome of the microhymenopteran Trichogramma brassicae were constructed from the analysis of segregation of random amplified polymorphic DNA markers in three F2 populations. These populations were composed of the haploid male progeny of several virgin F1 females, which resulted from the breeding of four parental lines that were nearly fixed for different random amplified polymorphic DNA markers and that were polymorphic for longevity and fecundity characters. As the order of markers common to the three mapping populations was found to be well conserved, a composite linkage map was constructed. Eighty-four markers were organized into five linkage groups and two pairs. The mean interval between two markers was 17.7 cM, and the map spanned 1330 cM. PMID- 9725847 TI - Overexpression Beadex mutations and loss-of-function heldup-a mutations in Drosophila affect the 3' regulatory and coding components, respectively, of the Dlmo gene. AB - LIM domains function as bridging modules between different members of multiprotein complexes. We report the cloning of a LIM-containing gene from Drosophila, termed Dlmo, which is highly homologous to the vertebrate LIM-only (LMO) genes. The 3' untranslated (UTR) of Dlmo contains multiple motifs implicated in negative post-transcriptional regulation, including AT-rich elements and Brd-like boxes. Dlmo resides in polytene band 17C1-2, where Beadex (Bx) and heldup-a (hdp-a) mutations map. We demonstrate that Bx mutations disrupt the 3'UTR of Dlmo, and thereby abrogate the putative negative control elements. This results in overexpression of Dlmo, which causes the wing scalloping that is typical of Bx mutants. We show that the erect wing phenotype of hdp-a results from disruption of the coding region of Dlmo. This provides molecular grounds for the suppression of the Bx phenotype by hdp-a mutations. Finally, we demonstrate phenotypic interaction between the LMO gene Dlmo, the LIM homeodomain gene apterous, and the Chip gene, which encodes a homolog of the vertebrate LIM interacting protein NLI/Ldb1. We propose that in analogy to their vertebrate counterparts, these proteins form a DNA-binding complex that regulates wing development. PMID- 9725848 TI - Detection of the ongoing sorting of ancestrally polymorphic SINEs toward fixation or loss in populations of two species of charr during speciation. AB - The FokI family of short interspersed repetitive elements (SINEs) has been found only in the genomes of charr fishes (genus Salvelinus). In an analysis of the insertion of FokI SINEs using PCR, we characterized six loci at which FokI SINEs have been inserted into the genomes of Salvelinus alpinus (Arctic charr) and/or S. malma (Dolly Varden). An analysis of one locus (Fok-223) suggested that a sister relationship exists between S. alpinus and S. malma and the SINE at this locus might have been inserted in a common ancestor of these two species, being fixed in all extant populations examined. By contrast, SINEs at two other loci (Fok-211 and Fok-206) were present specifically in the genome of S. alpinus, with polymorphism among populations of this species. Moreover, the presence or absence of the SINEs of the other three loci (Fok-214, Fok-217, and Fok-600) varied among populations of these two species. The most plausible interpretation of this result is that SINEs, which were ancestrally polymorphic in the genome of a common ancestor of these two species, are involved in an ongoing process of differential sorting and subsequent fixation in the various populations of each species. PMID- 9725849 TI - The complete nucleotide sequence of a snake (Dinodon semicarinatus) mitochondrial genome with two identical control regions. AB - The 17,191-bp mitochondrial DNA (mtDNA) of a Japanese colubrid snake, akamata (Dinodon semicarinatus), was cloned and sequenced. The snake mtDNA has some peculiar features that were found in our previous study using polymerase chain reaction: duplicate control regions that have completely identical sequences over 1 kbp, translocation of tRNALeu(UUR) gene, shortened TpsiC arm for most tRNA genes, and a pseudogene for tRNAPro. Phylogenetic analysis of amino acid sequences of protein genes suggested an unusually high rate of molecular evolution in the snake compared to other vertebrates. Southern hybridization experiments using mtDNAs purified from multiple akamata individuals showed that the duplicate state of the control region is not a transient or unstable feature found in a particular individual, but that it stably occurs in mitochondrial genomes of the species. This may, therefore, be regarded as an unprecedented example of stable functional redundancy in animal mtDNA. However, some of the examined individuals contain a rather scanty proportion of heteroplasmic mtDNAs with an organization of genes distinct from that of the major mtDNA. The gene organization of the minor mtDNA is in agreement with one of models that we present to account for the concerted evolution of duplicate control regions. PMID- 9725850 TI - The complete nucleotide sequence of the mitochondrial DNA of the dogfish, Scyliorhinus canicula. AB - We have determined the complete nucleotide sequence of the mitochondrial DNA (mtDNA) of the dogfish, Scyliorhinus canicula. The 16,697-bp-long mtDNA possesses a gene organization identical to that of the Osteichthyes, but different from that of the sea lamprey Petromyzon marinus. The main features of the mtDNA of osteichthyans were thus established in the common ancestor to chondrichthyans and osteichthyans. The phylogenetic analysis confirms that the Chondrichthyes are the sister group of the Osteichthyes. PMID- 9725851 TI - Molecular evolution of two lineages of L1 (LINE-1) retrotransposons in the california mouse, Peromyscus californicus. AB - The large number of L1 [long interspersed elements (LINE)-1] sequences found in the genome is due to the insertion of copies of the retrotransposon over evolutionary time. The majority of copies appear to be replicates of a few active, or "master" templates. A continual replacement of master templates over time gives rise to lineages distinguishable by their own unique set of shared sequence variants. A previous analysis of L1 sequences in deer mice, Peromyscus maniculatus and P. leucopus, revealed two active L1 lineages, marked by different rates of evolution, whose most recent common ancestor predates the expansion of the Peromyscus species. Here we exploit lineage-specific, shared-sequence variants to reveal a paucity of Lineage 2 sequences in at least one species, P. californicus. The dearth of Lineage 2 copies in P. californicus suggests that Lineage 2 may have been unproductive until after the most recent common ancestor of P. californicus and P. maniculatus. We also show that Lineage 1 appears to have a higher rate of evolution in P. maniculatus relative to either P. californicus or P. leucopus. As a phylogenetic tool, L1 lineage-specific variants support a close affinity between P. californicus and P. eremicus relative to the other species examined. PMID- 9725853 TI - Quantitative trait loci affecting body weight and fatness from a mouse line selected for extreme high growth. AB - Quantitative trait loci (QTL) influencing body weight were mapped by linkage analysis in crosses between a high body weight selected line (DU6) and a control line (DUKs). The two mouse lines differ in body weight by 106% and in abdominal fat weight by 100% at 42 days. They were generated from the same base population and maintained as outbred colonies. Determination of line-specific allele frequencies at microsatellite markers spanning the genome indicated significant changes between the lines on 15 autosomes and the X chromosome. To confirm these effects, a QTL analysis was performed using structured F2 pedigrees derived from crosses of a single male from DU6 with a female from DUKs. QTL significant at the genome-wide level were mapped for body weight on chromosome 11; for abdominal fat weight on chromosomes 4, 11, and 13; for abdominal fat percentage on chromosomes 3 and 4; and for the weights of liver on chromosomes 4 and 11, of kidney on chromosomes 2 and 9, and of spleen on chromosome 11. The strong effect on body weight of the QTL on chromosome 11 was confirmed in three independent pedigrees. The effect was additive and independent of sex, accounting for 21-35% of the phenotypic variance of body weight within the corresponding F2 populations. The test for multiple QTL on chromosome 11 with combined data from all pedigrees indicated the segregation of two loci separated by 36 cM influencing body weight. PMID- 9725852 TI - Deleterious mutations at the mitochondrial ND3 gene in South American marsh rats (Holochilus). AB - Statistical analyses of DNA sequences have revealed patterns of nonneutral evolution in mitochondrial DNA of mice, humans, and Drosophila. Here we report patterns of mitochondrial sequence evolution in South American marsh rats (genus Holochilus). We sequenced the complete mitochondrial ND3 gene in 82 Holochilus brasiliensis and 21 H. vulpinus to test the neutral prediction that the ratio of nonsynonymous to synonymous nucleotide changes is the same within and between species. Within H. brasiliensis we observed a greater number of amino acid polymorphisms than expected based on interspecific comparisons. This contingency table analysis suggests that many amino acid polymorphisms are mildly deleterious. Several tests of the frequency distribution also revealed departures from a neutral, equilibrium model, and these departures were observed for both nonsynonymous and synonymous sites. In general, an excess of rare sites was observed, consistent with either a recent selective sweep or with populations not at mutation-drift equilibrium. PMID- 9725854 TI - Cosegregation of single genes associated with fertility restoration and transcript processing of sorghum mitochondrial orf107 and urf209. AB - Defective nuclear-cytoplasmic interactions leading to aberrant microgametogenesis in sorghum carrying the IS1112C male-sterile cytoplasm occur very late in pollen maturation. Amelioration of this condition, the restoration of pollen viability, involves a novel two-gene gametophytic system, wherein genes designated Rf3 and Rf4 are required for viability of individual gametes. Rf3 is tightly linked to, or represents, a single gene that regulates a transcript processing activity that cleaves transcripts of orf107, a chimeric mitochondrial open reading frame specific to IS1112C. The mitochondrial gene urf209 is also subject to nucleus specific enhanced transcript processing, 5' to the gene, conferred by a single dominant gene designated Mmt1. Examinations of transcript patterns in F2 and two backcross populations indicated cosegregation of the augmented orf107 and urf209 processing activities in IS1112C. Several sorghum lines that do not restore fertility or confer orf107 transcript processing do exhibit urf209 transcript processing, indicating that the activities are distinguishable. We conclude that the nuclear gene(s) conferring enhanced orf107 and urf209 processing activities are tightly linked in IS1112C. Alternatively, the similarity in apparent regulatory action of the genes may indicate allelic differences wherein the IS1112C Rf3 allele may differ from alleles of maintainer lines by the capability to regulate both orf107 and urf209 processing activities. PMID- 9725855 TI - The association of flowering time quantitative trait loci with duplicated regions and candidate loci in Brassica oleracea. AB - A population of 150 doubled haploid lines of rapid cycling Brassica oleracea, derived from an F1 from a var. alboglabra x var. italica cross, was scored for flowering time in two trials. Using information on 82 mapped molecular markers, spread evenly across the nine linkage groups, QTL were identified at six locations; one each on linkage groups O2 and O3 and two each on linkage groups O5 and O9. In total, these QTL explained 58 and 93% of the genetical variation in the two trials. Three of these QTL, on linkage groups O2, O3, and O9, were situated in regions showing considerable homology both with each other and with chromosome regions of B. nigra that have been shown to affect flowering time. These same regions are all homologous to a single tract of Arabidopsis chromosome 5, which contains a number of the flowering-related genes, one or more of which may be candidates for the QTL found in Brassica. PMID- 9725856 TI - Flowering-time genes modulate the response to LEAFY activity. AB - Among the genes that control the transition to flowering in Arabidopsis is a large group whose inactivation causes a delay in flowering. It has been difficult to establish different pathways in which the flowering-time genes might act, because mutants with lesions in these genes have very similar phenotypes. Among the putative targets of the flowering-time genes is another group of genes, which control the identity of individual meristems. Overexpression of one of the meristem-identity genes, LEAFY, can cause the precocious generation of flowers and thus early flowering. We have exploited the opposite phenotypes seen in late flowering mutants and LEAFY overexpressers to clarify the genetic interactions between flowering-time genes and LEAFY. According to epistatic relationships, we can define one class of flowering-time genes that affects primarily the response to LEAFY activity, and another class of genes that affects primarily the transcriptional induction of LEAFY. These observations allow us to expand previously proposed models for the genetic control of flowering time. PMID- 9725857 TI - Arabidopsis TSO1 regulates directional processes in cells during floral organogenesis. AB - Flowers of the previously described Arabidopsis tso1-1 mutant had aberrant, highly reduced organs in place of petals, stamens, and carpels. Cells of tso1-1 flowers had division defects, including failure in cytokinesis, partial cell wall formation, and elevated nuclear DNA content. We describe here two new tso1 alleles (tso1-3 and tso1-4), which caused defects in ovule development, but had little effect on gross floral morphology. Early ovule development occurred normally in tso1-3 and tso1-4, but the shapes and alignments of integument cells became increasingly more disordered as development progressed. tso1-3 ovules usually lacked embryo sacs due to a failure to form megaspore mother cells. The cell division defects described for the strong tso1-1 mutant were rarely observed in tso1-3 ovules. The aberrations in tso1-3 mutants primarily resulted from a failure in directional expansion of cells and/or coordination of this process among adjacent cells. Effects of tso1-3 appeared to be independent of effects of other ovule development mutations, with the exception of leunig, which exhibited a synergistic interaction. The data are consistent with TSO1 acting in processes governing directional movement of cellular components, indicating a likely role for TSO1 in cytoskeletal function. PMID- 9725858 TI - A subset of conserved tRNA genes in plastid DNA of nongreen plants. AB - The plastid genome of the nonphotosynthetic parasitic plant Epifagus virginiana contains only 17 of the 30 tRNA genes normally found in angiosperm plastid DNA. Although this is insufficient for translation, the genome is functional, so import of cytosolic tRNAs into plastids has been suggested. This raises the question of whether the tRNA genes that remain in E. virginiana plastid DNA are active or have just fortuitously escaped deletion. We report the sequences of 20 plastid tRNA loci from Orobanche minor, which shares a nonphotosynthetic ancestor with E. virginiana. The two species have 9 intact tRNA genes in common, the others being defunct in one or both species. The intron-containing trnLUAA gene is absent from E. virginiana, but it is intact, transcribed, and spliced in O. minor. The shared intact genes are better conserved than intergenic sequences, which indicates that these genes are being maintained by natural selection and, therefore, must be functional. For the most part, the tRNA species conserved in nonphotosynthetic plastids are also those that have never been found to be imported in plant mitochondria, which suggests that the same rules may govern tRNA import in the two organelles. A small photosynthesis gene, psbI, is still intact in O. minor, and computer simulations show that some small nonessential genes have an appreciable chance of escaping deletion. PMID- 9725859 TI - Bottleneck effect on genetic variance. A theoretical investigation of the role of dominance. AB - The phenomenon that the genetic variance of fitness components increase following a bottleneck or inbreeding is supported by a growing number of experiments and is explained theoretically by either dominance or epistasis. In this article, diffusion approximations under the infinite sites model are used to quantify the effect of dominance, using data on viability in Drosophila melanogaster. The model is based on mutation parameters from mutation accumulation experiments involving balancer chromosomes (set I) or inbred lines (set II). In essence, set I assumes many mutations of small effect, whereas set II assumes fewer mutations of large effect. Compared to empirical estimates from large outbred populations, set I predicts reasonable genetic variances but too low mean viability. In contrast, set II predicts a reasonable mean viability but a low genetic variance. Both sets of parameters predict the changes in mean viability (depression), additive variance, between-line variance and heritability following bottlenecks generally compatible with empirical results, and these changes are mainly caused by lethals and deleterious mutants of large effect. This article suggests that dominance is the main cause for increased genetic variances for fitness components and fitness-related traits after bottlenecks observed in various experiments. PMID- 9725860 TI - The population genetics of synthetic lethals. AB - Synthetic lethals are variants at different loci that have little or no effect on viability singly but cause lethality in combination. The importance of synthetic lethals and, more generally, of synthetic deleterious loci (SDL) has been controversial. Here, we derive the expected frequencies for SDL under a mutation selection balance for the complete haploid model and selected cases of the diploid model. We have also obtained simple approximations that demonstrate good fit to exact solutions based on numerical iterations. In the haploid case, equilibrium frequencies of carrier haplotypes (individuals with only a single mutation) are comparable to analogous single-locus results, after allowing for the effects of linkage. Frequencies in the diploid case, however, are much higher and more comparable to the square root of the single-locus results. In particular, when selection operates only on the double-mutant homozygote and linkage is not too tight, the expected frequency of the carriers is approximately the quartic root of the ratio between the mutation rate and the selection coefficient of the synthetics. For a reasonably wide set of models, the frequencies of carriers can be on the order of a few percent. The equilibrium frequencies of these deleterious alleles can be relatively high because, with SDL, both dominance and epistasis act to shield carriers from exposure to selection. We also discuss the possible role of SDL in maintaining genetic variation and in hybrid breakdown. PMID- 9725861 TI - Statistical analysis of ordered tetrads. AB - Ordered tetrad data yield information on chromatid interference, chiasma interference, and centromere locations. In this article, we show that the assumption of no chromatid interference imposes certain constraints on multilocus ordered tetrad probabilities. Assuming no chromatid interference, these constraints can be used to order markers under general chiasma processes. We also derive multilocus tetrad probabilities under a class of chiasma interference models, the chi-square models. Finally, we compare centromere map functions under the chi-square models with map functions proposed in the literature. Results in this article can be applied to order genetic markers and map centromeres using multilocus ordered tetrad data. PMID- 9725862 TI - Statistical analysis of half-tetrads. AB - Half-tetrads, where two meiotic products from a single meiosis are recovered together, arise in different forms in a variety of organisms. Closely related to ordered tetrads, half-tetrads yield information on chromatid interference, chiasma interference, and centromere positions. In this article, for different half-tetrad types and different marker configurations, we derive the relations between multilocus half-tetrad probabilities and multilocus ordered tetrad probabilities. These relations are used to obtain equality and inequality constraints among multilocus half-tetrad probabilities that are imposed by the assumption of no chromatid interference. We illustrate how to apply these results to study chiasma interference and to map centromeres using multilocus half-tetrad data. PMID- 9725863 TI - Simultaneous estimation of all the parameters of a stepwise mutation model. AB - Minisatellite and microsatellite are short tandemly repetitive sequences dispersed in eukaryotic genomes, many of which are highly polymorphic due to copy number variation of the repeats. Because mutation changes copy numbers of the repeat sequences in a generalized stepwise fashion, stepwise mutation models are widely used for studying the dynamics of these loci. We propose a minimum chi square (MCS) method for simultaneous estimation of all the parameters in a stepwise mutation model and the ancestral allelic type of a sample. The MCS estimator requires knowing the mean number of alleles of a certain size in a sample, which can be estimated using Monte Carlo samples generated by a coalescent algorithm. The method is applied to samples of seven (CA)n repeat loci from eight human populations and one chimpanzee population. The estimated values of parameters suggest that there is a general tendency for microsatellite alleles to expand in size, because (1) each mutation has a slight tendency to cause size increase and (2) the mean size increase is larger than the mean size decrease for a mutation. Our estimates also suggest that most of these CA-repeat loci evolve according to multistep mutation models rather than single-step mutation models. We also introduced several quantities for measuring the quality of the estimation of ancestral allelic type, and it appears that the majority of the estimated ancestral allelic types are reasonably accurate. Implications of our analysis and potential extensions of the method are discussed. SINCE the discovery that a large number of loci with tandemly repeated sequences in human and many eukaryote species are highly polymorphic because of copy number variation of the repeats in different individuals (Jeffreys 1985; Litt and Luty 1989; Weber and May 1989), allele size data from such loci are rapidly becoming the dominant source of genetic markers for genome mapping, forensic testing, and population studies. Loci with repeat sequences longer than 5 bp are generally referred to as minisatellite or variable number tandem repeat loci, and those with repeat sequences between 2 to 5 bp are referred to as microsatellite or short tandem repeat loci (Tautz 1993). Because mutations change the copy number of such loci in a stepwise fashion, rapid accumulation of population samples from minisatellite and microsatellite loci has resurrected the interest of the stepwise mutation model (SMM), which was popular in the 1970s. PMID- 9725864 TI - Genealogical inference from microsatellite data. AB - Ease and accuracy of typing, together with high levels of polymorphism and widespread distribution in the genome, make microsatellite (or short tandem repeat) loci an attractive potential source of information about both population histories and evolutionary processes. However, microsatellite data are difficult to interpret, in particular because of the frequency of back-mutations. Stochastic models for the underlying genetic processes can be specified, but in the past they have been too complicated for direct analysis. Recent developments in stochastic simulation methodology now allow direct inference about both historical events, such as genealogical coalescence times, and evolutionary parameters, such as mutation rates. A feature of the Markov chain Monte Carlo (MCMC) algorithm that we propose here is that the likelihood computations are simplified by treating the (unknown) ancestral allelic states as auxiliary parameters. We illustrate the algorithm by analyzing microsatellite samples simulated under the model. Our results suggest that a single microsatellite usually does not provide enough information for useful inferences, but that several completely linked microsatellites can be informative about some aspects of genealogical history and evolutionary processes. We also reanalyze data from a previously published human Y chromosome microsatellite study, finding evidence for an effective population size for human Y chromosomes in the low thousands and a recent time since their most recent common ancestor: the 95% interval runs from approximately 15, 000 to 130,000 years, with most likely values around 30,000 years. PMID- 9725888 TI - Exploring local calcium feedback: trying to fool mother nature. PMID- 9725889 TI - Effects of partial sarcoplasmic reticulum calcium depletion on calcium release in frog cut muscle fibers equilibrated with 20 mM EGTA. AB - Resting sarcoplasmic reticulum (SR) Ca content ([CaSR]R) was varied in cut fibers equilibrated with an internal solution that contained 20 mM EGTA and 0-1.76 mM Ca. SR Ca release and [CaSR]R were measured with the EGTA-phenol red method (. J. Gen. Physiol. 106:259-336). After an action potential, the fractional amount of Ca released from the SR increased from 0.17 to 0.50 when [CaSR]R was reduced from 1, 200 to 140 microM. This increase was associated with a prolongation of release (final time constant, from 1-2 to 10-15 ms) and of the action potential (by 1-2 ms). Similar changes in release were observed with brief stimulations to -20 mV in voltage-clamped fibers, in which charge movement (Qcm) could be measured. The peak values of Qcm and the fractional rate of SR Ca release, as well as their ON time courses, were little affected by reducing [CaSR]R from 1,200 to 140 microM. After repolarization, however, the OFF time courses of Qcm and the rate of SR Ca release were slowed by factors of 1.5-1.7 and 6.5, respectively. These and other results suggest that, after action potential stimulation of fibers in normal physiological condition, the increase in myoplasmic free [Ca] that accompanies SR Ca release exerts three negative feedback effects that tend to reduce additional release: (a) the action potential is shortened by current through Ca-activated potassium channels in the surface and/or tubular membranes; (b) the OFF kinetics of Qcm is accelerated; and (c) Ca inactivation of Ca release is increased. Some of these effects of Ca on an SR Ca channel or its voltage sensor appear to be regulated by the value of [Ca] within 22 nm of the mouth of the channel. PMID- 9725890 TI - Model of sarcomeric Ca2+ movements, including ATP Ca2+ binding and diffusion, during activation of frog skeletal muscle. AB - Cannell and Allen (1984. Biophys. J. 45:913-925) introduced the use of a multi compartment model to estimate the time course of spread of calcium ions (Ca2+) within a half sarcomere of a frog skeletal muscle fiber activated by an action potential. Under the assumption that the sites of sarcoplasmic reticulum (SR) Ca2+ release are located radially around each myofibril at the Z line, their model calculated the spread of released Ca2+ both along and into the half sarcomere. During diffusion, Ca2+ was assumed to react with metal-binding sites on parvalbumin (a diffusible Ca2+- and Mg2+-binding protein) as well as with fixed sites on troponin. We have developed a similar model, but with several modifications that reflect current knowledge of the myoplasmic environment and SR Ca2+ release. We use a myoplasmic diffusion constant for free Ca2+ that is twofold smaller and an SR Ca2+ release function in response to an action potential that is threefold briefer than used previously. Additionally, our model includes the effects of Ca2+ and Mg2+ binding by adenosine 5'-triphosphate (ATP) and the diffusion of Ca2+-bound ATP (CaATP). Under the assumption that the total myoplasmic concentration of ATP is 8 mM and that the amplitude of SR Ca2+ release is sufficient to drive the peak change in free [Ca2+] (Delta[Ca2+]) to 18 microM (the approximate spatially averaged value that is observed experimentally), our model calculates that (a) the spatially averaged peak increase in [CaATP] is 64 microM; (b) the peak saturation of troponin with Ca2+ is high along the entire thin filament; and (c) the half-width of Delta[Ca2+] is consistent with that observed experimentally. Without ATP, the calculated half-width of spatially averaged Delta[Ca2+] is abnormally brief, and troponin saturation away from the release sites is markedly reduced. We conclude that Ca2+ binding by ATP and diffusion of CaATP make important contributions to the determination of the amplitude and the time course of Delta[Ca2+]. PMID- 9725891 TI - The diphtheria toxin channel-forming T domain translocates its own NH2-terminal region across planar bilayers. AB - The T domain of diphtheria toxin, which extends from residue 202 to 378, causes the translocation of the catalytic A fragment (residues 1-201) across endosomal membranes and also forms ion-conducting channels in planar phospholipid bilayers. The carboxy terminal 57-amino acid segment (322-378) in the T domain is all that is required to form these channels, but its ability to do so is greatly augmented by the portion of the T domain upstream from this. In this work, we show that in association with channel formation by the T domain, its NH2 terminus, as well as some or all of the adjacent hydrophilic 63 amino acid segment, cross the lipid bilayer. The phenomenon that enabled us to demonstrate that the NH2-terminal region of the T domain was translocated across the membrane was the rapid closure of channels at cis negative voltages when the T domain contained a histidine tag at its NH2 terminus. The inhibition of this effect by trans nickel, and by trans streptavidin when the histidine tag sequence was biotinylated, clearly established that the histidine tag was present on the trans side of the membrane. Furthermore, the inhibition of rapid channel closure by trans trypsin, combined with mutagenesis to localize the trypsin site, indicated that some portion of the 63 amino acid NH2-terminal segment of the T domain was also translocated to the trans side of the membrane. If the NH2 terminus was forced to remain on the cis side, by streptavidin binding to the biotinylated histidine tag sequence, channel formation was severely disrupted. Thus, normal channel formation by the T domain requires that its NH2 terminus be translocated across the membrane from the cis to the trans side, even though the NH2 terminus is >100 residues removed from the channel-forming part of the molecule. PMID- 9725892 TI - Mechanism of ATP-sensitive K channel inhibition by sulfhydryl modification. AB - ATP-sensitive potassium (KATP) channels are reversibly inhibited by intracellular ATP. Agents that interact with sulfhydryl moieties produce an irreversible inhibition of KATP channel activity when applied to the intracellular membrane surface. ATP appears to protect against this effect, suggesting that the cysteine residue with which thiol reagents interact may either lie within the ATP-binding site or be inaccessible when the channel is closed. We have examined the interaction of the membrane-impermeant thiol-reactive agent p chloromercuriphenylsulphonate (pCMPS) with the cloned beta cell KATP channel. This channel comprises the pore-forming Kir6.2 and regulatory SUR1 subunits. We show that the cysteine residue involved in channel inhibition by pCMPS resides on the Kir6.2 subunit and is located at position 42, which lies within the NH2 terminus of the protein. Although ATP protects against the effects of pCMPS, the ATP sensitivity of the KATP channel was unchanged by mutation of C42 to either valine (V) or alanine (A), suggesting that ATP does not interact directly with this residue. These results are consistent with the idea that C42 is inaccessible to the intracellular solution, and thereby protected from interaction with pCMPS when the channel is closed by ATP. We also observed that the C42A mutation does not affect the ability of SUR1 to endow Kir6.2 with diazoxide sensitivity, and reduces, but does not prevent, the effects of MgADP and tolbutamide, which are mediated via SUR1. The Kir6.2-C42A (or V) mutant channel may provide a suitable background for cysteine-scanning mutagenesis studies. PMID- 9725893 TI - Molecular analysis of ATP-sensitive K channel gating and implications for channel inhibition by ATP. AB - The beta cell KATP channel is an octameric complex of four pore-forming subunits (Kir6.2) and four regulatory subunits (SUR1). A truncated isoform of Kir6.2 (Kir6.2DeltaC26), which expresses independently of SUR1, shows intrinsic ATP sensitivity, suggesting that this subunit is primarily responsible for mediating ATP inhibition. We show here that mutation of C166, which lies at the cytosolic end of the second transmembrane domain, to serine (C166S) increases the open probability of Kir6.2DeltaC26 approximately sevenfold by reducing the time the channel spends in a long closed state. Rundown of channel activity is also decreased. Kir6.2DeltaC26 containing the C166S mutation shows a markedly reduced ATP sensitivity: the Ki is reduced from 175 microM to 2.8 mM. Substitution of threonine, alanine, methionine, or phenylalanine at position C166 also reduced the channel sensitivity to ATP and simultaneously increased the open probability. Thus, ATP does not act as an open channel blocker. The inhibitory effects of tolbutamide are reduced in channels composed of SUR1 and Kir6.2 carrying the C166S mutation. Our results are consistent with the idea that C166 plays a role in the intrinsic gating of the channel, possibly by influencing a gate located at the intracellular end of the pore. Kinetic analysis suggests that the apparent decrease in ATP sensitivity, and the changes in other properties, observed when C166 is mutated is largely a consequence of the impaired transition from the open to the long closed state. PMID- 9725895 TI - Criteria for the molecular identification of the volume-sensitive outwardly rectifying Cl- channel. PMID- 9725894 TI - Block of the Kir2.1 channel pore by alkylamine analogues of endogenous polyamines. AB - Inward rectification induced by mono- and diaminoalkane application to inside-out membrane patches was studied in Kir2.1 (IRK1) channels expressed in Xenopus oocytes. Both monoamines and diamines block Kir2.1 channels, with potency increasing as the alkyl chain length increases (from 2 to 12 methylene groups), indicating a strong hydrophobic interaction with the blocking site. For diamines, but not monoamines, increasing the alkyl chain also increases the steepness of the voltage dependence, at any concentration, from a limiting minimal value of approximately 1.5 (n = 2 methylene groups) to approximately 4 (n = 10 methylene groups). These observations lead us to hypothesize that monoamines and diamines block inward rectifier K+ channels by entering deeply into a long, narrow pore, displacing K+ ions to the outside of the membrane, with this displacement of K+ ions contributing to "extra" charge movement. All monoamines are proposed to lie with the "head" amine at a fixed position in the pore, determined by electrostatic interaction, so that zdelta is independent of monoamine alkyl chain length. The head amine of diamines is proposed to lie progressively further into the pore as alkyl chain length increases, thus displacing more K+ ions to the outside, resulting in charge movement (zdelta) increasing with the increase in alkyl chain length. PMID- 9725896 TI - Identifying swelling-activated channels from ion selectivity patterns. PMID- 9725898 TI - An alpha-tubulin mutant destabilizes the heterodimer: phenotypic consequences and interactions with tubulin-binding proteins. AB - Many effectors of microtubule assembly in vitro enhance the polymerization of subunits. However, several Saccharomyces cerevisiae genes that affect cellular microtubule-dependent processes appear to act at other steps in assembly and to affect polymerization only indirectly. Here we use a mutant alpha-tubulin to probe cellular regulation of microtubule assembly. tub1-724 mutant cells arrest at low temperature with no assembled microtubules. The results of several assays reported here demonstrate that the heterodimer formed between Tub1-724p and beta tubulin is less stable than wild-type heterodimer. The unstable heterodimer explains several conditional phenotypes conferred by the mutation. These include the lethality of tub1-724 haploid cells when the beta-tubulin-binding protein Rbl2p is either overexpressed or absent. It also explains why the TUB1/tub1-724 heterozygotes are cold sensitive for growth and why overexpression of Rbl2p rescues that conditional lethality. Both haploid and heterozygous tub1-724 cells are inviable when another microtubule effector, PAC2, is overexpressed. These effects are explained by the ability of Pac2p to bind alpha-tubulin, a complex we demonstrate directly. The results suggest that tubulin-binding proteins can participate in equilibria between the heterodimer and its components. PMID- 9725897 TI - The role of preassembled cytoplasmic complexes in assembly of flagellar dynein subunits. AB - Previous work has revealed a cytoplasmic pool of flagellar precursor proteins capable of contributing to the assembly of new flagella, but how and where these components assemble is unknown. We tested Chlamydomonas outer-dynein arm subunit stability and assembly in the cytoplasm of wild-type cells and 11 outer dynein arm assembly mutant strains (oda1-oda11) by Western blotting of cytoplasmic extracts, or immunoprecipitates from these extracts, with five outer-row dynein subunit-specific antibodies. Western blots reveal that at least three oda mutants (oda6, oda7, and oda9) alter the level of a subunit that is not the mutant gene product. Immunoprecipitation shows that large preassembled flagellar complexes containing all five tested subunits (three heavy chains and two intermediate chains) exist within wild-type cytoplasm. When the preassembly of these subunits was examined in oda strains, we observed three patterns: complete coassembly (oda 1, 3, 5, 8, and 10), partial coassembly (oda7 and oda11), and no coassembly (oda2, 6, and 9) of the four tested subunits with HCbeta. Our data, together with previous studies, suggest that flagellar outer-dynein arms preassemble into a complete Mr approximately 2 x 10(6) dynein arm that resides in a cytoplasmic precursor pool before transport into the flagellar compartment. PMID- 9725899 TI - Characterization of ATM expression, localization, and associated DNA-dependent protein kinase activity. AB - Ataxia telangiectasia-mutated gene (ATM) is a 350-kDa protein whose function is defective in the autosomal recessive disorder ataxia telangiectasia (AT). Affinity-purified polyclonal antibodies were used to characterize ATM. Steady state levels of ATM protein varied from undetectable in most AT cell lines to highly expressed in HeLa, U2OS, and normal human fibroblasts. Subcellular fractionation showed that ATM is predominantly a nuclear protein associated with the chromatin and nuclear matrix. ATM protein levels remained constant throughout the cell cycle and did not change in response to serum stimulation. Ionizing radiation had no significant effect on either the expression or distribution of ATM. ATM immunoprecipitates from HeLa cells and the human DNA-dependent protein kinase null cell line MO59J, but not from AT cells, phosphorylated the 34-kDa subunit of replication protein A (RPA) complex in a single-stranded and linear double-stranded DNA-dependent manner. Phosphorylation of p34 RPA occurred on threonine and serine residues. Phosphopeptide analysis demonstrates that the ATM associated protein kinase phosphorylates p34 RPA on similar residues observed in vivo. The DNA-dependent protein kinase activity observed for ATM immunocomplexes, along with the association of ATM with chromatin, suggests that DNA damage can induce ATM or a stably associated protein kinase to phosphorylate proteins in the DNA damage response pathway. PMID- 9725900 TI - Evidence for the presence of 5S rRNA in mammalian mitochondria. AB - Mammalian mitochondrial ribosomes contain two prokaryotic-like rRNAs, 12S and 16S, both encoded by mitochondrial DNA. As opposed to cytosolic ribosomes, however, these ribosomes are not thought to contain 5S rRNA. For this reason, it has been unclear whether 5S rRNA, which can be detected in mitochondrial preparations, is an authentic organellar species imported from the cytosol or is merely a copurifying cytosol-derived contaminant. We now show that 5S rRNA is tightly associated with highly purified mitochondrial fractions of human and rat cells and that 5S rRNA transcripts derived from a synthetic gene transfected transiently into human cells are both expressed in vivo and present in highly purified mitochondria and mitoplasts. We conclude that 5S rRNA is imported into mammalian mitochondria, but its function there still remains to be clarified. PMID- 9725901 TI - Binding of insulin-like growth factor (IGF)-binding protein-5 to smooth-muscle cell extracellular matrix is a major determinant of the cellular response to IGF I. AB - Insulin-like growth factor-binding protein-5 (IGFBP-5) has been shown to bind to fibroblast extracellular matrix (ECM). Extracellular matrix binding of IGFBP-5 leads to a decrease in its affinity for insulin-like growth factor-I (IGF-I), which allows IGF-I to better equilibrate with IGF receptors. When the amount of IGFBP-5 that is bound to ECM is increased by exogenous addition, IGF-I's effect on fibroblast growth is enhanced. In this study we identified the specific basic residues in IGFBP-5 that mediate its binding to porcine smooth-muscle cell (pSMC) ECM. An IGFBP-5 mutant containing alterations of basic residues at positions 211, 214, 217, and 218 had the greatest reduction in ECM binding, although three other mutants, R214A, R207A/K211N, and K202A/R206N/R207A, also had major decreases. In contrast, three other mutants, R201A/K202N/R206N/R208A, and K217N/R218A and K211N, had only minimal reductions in ECM binding. This suggested that residues R207 and R214 were the most important for binding, whereas alterations in K211 and R218, which align near them, had minimal effects. To determine the effect of a reduction in ECM binding on the cellular replication response to IGF-I, pSMCs were transfected with the mutant cDNAs that encoded the forms of IGFBPs with the greatest changes in ECM binding. The ECM content of IGFBP-5 from cultures expressing the K211N, R214A, R217A/R218A, and K202A/R206N/R207A mutants was reduced by 79.6 and 71.7%, respectively, compared with cells expressing the wild type protein. In contrast, abundance of the R201A/K202N/R206N/R208A mutant was reduced by only 14%. Cells expressing the two mutants with reduced ECM binding had decreased DNA synthesis responses to IGF-I, but the cells expressing the R201A/K202N/R206N/R208A mutant responded well to IGF-I. The findings suggest that specific basic amino acids at positions 207 and 214 mediate the binding of IGFBP 5 to pSMC/ECM. Smooth-muscle cells that constitutively express the mutants that bind weakly to ECM are less responsive to IGF-I, suggesting that ECM binding of IGFBP-5 is an important variable that determines cellular responsiveness. PMID- 9725902 TI - Phosphorylation of sic1, a cyclin-dependent kinase (Cdk) inhibitor, by Cdk including Pho85 kinase is required for its prompt degradation. AB - In the yeast Saccharomyces cerevisiae, Sic1, an inhibitor of Clb-Cdc28 kinases, must be phosphorylated and degraded in G1 for cells to initiate DNA replication, and Cln-Cdc28 kinase appears to be primarily responsible for phosphorylation of Sic1. The Pho85 kinase is a yeast cyclin-dependent kinase (Cdk), which is not essential for cell growth unless both CLN1 and CLN2 are absent. We demonstrate that Pho85, when complexed with Pcl1, a G1 cyclin homologue, can phosphorylate Sic1 in vitro, and that Sic1 appears to be more stable in pho85Delta cells. Three consensus Cdk phosphorylation sites present in Sic1 are phosphorylated in vivo, and two of them are required for prompt degradation of the inhibitor. Pho85 and other G1 Cdks appear to phosphorylate Sic1 at different sites in vivo. Thus at least two distinct Cdks can participate in phosphorylation of Sic1 and may therefore regulate progression through G1. PMID- 9725903 TI - Partially processed pre-rRNA is preserved in association with processing components in nucleolus-derived foci during mitosis. AB - Previous studies showed that components implicated in pre-rRNA processing, including U3 small nucleolar (sno)RNA, fibrillarin, nucleolin, and proteins B23 and p52, accumulate in perichromosomal regions and in numerous mitotic cytoplasmic particles, termed nucleolus-derived foci (NDF) between early anaphase and late telophase. The latter structures were analyzed for the presence of pre rRNA by fluorescence in situ hybridization using probes for segments of pre-rRNA with known half-lives. The NDF did not contain the short-lived 5'-external transcribed spacer (ETS) leader segment upstream from the primary processing site in 47S pre-rRNA. However, the NDF contained sequences from the 5'-ETS core, 18S, internal transcribed spacer 1 (ITS1), and 28S segments and also had detectable, but significantly reduced, levels of the 3'-ETS sequence. Northern analyses showed that in mitotic cells, the latter sequences were present predominantly in 45S-46S pre-rRNAs, indicating that high-molecular weight processing intermediates are preserved during mitosis. Two additional essential processing components were also found in the NDF: U8 snoRNA and hPop1 (a protein component of RNase MRP and RNase P). Thus, the NDF appear to be large complexes containing partially processed pre-rRNA associated with processing components in which processing has been significantly suppressed. The NDF may facilitate coordinated assembly of postmitotic nucleoli. PMID- 9725904 TI - A novel synaptobrevin/VAMP homologous protein (VAMP5) is increased during in vitro myogenesis and present in the plasma membrane. AB - cDNA clones encoding a novel protein (VAMP5) homologous to synaptobrevins/VAMPs are detected during database searches. The predicted 102-amino acid VAMP5 harbors a 23-residue hydrophobic region near the carboxyl terminus and exhibits an overall amino acid identity of 33% with synaptobrevin/VAMP1 and 2 and cellubrevin. Northern blot analysis reveals that the mRNA for VAMP5 is preferentially expressed in the skeletal muscle and heart, whereas significantly lower levels are detected in several other tissues but not in the brain. During in vitro differentiation (myogenesis) of C2C12 myoblasts into myotubes, the mRNA level for VAMP5 is increased approximately 8- to 10-fold. Immunoblot analysis using antibodies specific for VAMP5 shows that the protein levels are also elevated approximately 6-fold during in vitro myogenesis of C2C12 cells. Indirect immunofluorescence microscopy and immunoelectron microscopy reveal that VAMP5 is associated with the plasma membrane as well as intracellular perinuclear and peripheral vesicular structures of myotubes. Epitope-tagged versions of VAMP5 are similarly targeted to the plasma membrane. PMID- 9725905 TI - A novel fluorescence-based genetic strategy identifies mutants of Saccharomyces cerevisiae defective for nuclear pore complex assembly. AB - Nuclear pore complexes (NPCs) are large proteinaceous portals for exchanging macromolecules between the nucleus and the cytoplasm. Revealing how this transport apparatus is assembled will be critical for understanding the nuclear transport mechanism. To address this issue and to identify factors that regulate NPC formation and dynamics, a novel fluorescence-based strategy was used. This approach is based on the functional tagging of NPC proteins with the green fluorescent protein (GFP), and the hypothesis that NPC assembly mutants will have distinct GFP-NPC signals as compared with wild-type (wt) cells. By fluorescence activated cell sorting for cells with low GFP signal from a population of mutagenized cells expressing GFP-Nup49p, three complementation groups were identified: two correspond to mutant nup120 and gle2 alleles that result in clusters of NPCs. Interestingly, a third group was a novel temperature-sensitive allele of nup57. The lowered GFP-Nup49p incorporation in the nup57-E17 cells resulted in a decreased fluorescence level, which was due in part to a sharply diminished interaction between the carboxy-terminal truncated nup57pE17 and wt Nup49p. Interestingly, the nup57-E17 mutant also affected the incorporation of a specific subset of other nucleoporins into the NPC. Decreased levels of NPC associated Nsp1p and Nup116p were observed. In contrast, the localizations of Nic96p, Nup82p, Nup159p, Nup145p, and Pom152p were not markedly diminished. Coincidentally, nuclear import capacity was inhibited. Taken together, the identification of such mutants with specific perturbations of NPC structure validates this fluorescence-based strategy as a powerful approach for providing insight into the mechanism of NPC biogenesis. PMID- 9725906 TI - Real-time imaging of the axonal transport of granules containing a tissue plasminogen activator/green fluorescent protein hybrid. AB - A hybrid protein, tPA/GFP, consisting of rat tissue plasminogen activator (tPA) and green fluorescent protein (GFP) was expressed in PC12 cells and used to study the distribution, secretory behavior, and dynamics of secretory granules containing tPA in living cells with a neuronal phenotype. High-resolution images demonstrate that tPA/GFP has a growth cone-biased distribution in differentiated cells and that tPA/GFP is transported in granules of the regulated secretory pathway that colocalize with granules containing secretogranin II. Time-lapse images of secretion reveal that secretagogues induce substantial loss of cellular tPA/GFP fluorescence, most importantly from growth cones. Time-lapse images of the axonal transport of granules containing tPA/GFP reveal a surprising complexity to granule dynamics. Some granules undergo canonical fast axonal transport; others move somewhat more slowly, especially in highly fluorescent neurites. Most strikingly, granules traffic bidirectionally along neurites to an extent that depends on granule accumulation, and individual granules can reverse their direction of motion. The retrograde component of this bidirectional transport may help to maintain cellular homeostasis by transporting excess tPA/GFP back toward the cell body. The results presented here provide a novel view of the axonal transport of secretory granules. In addition, the results suggest that tPA is targeted for regulated secretion from growth cones of differentiated cells, strategically positioning tPA to degrade extracellular barriers or to activate other barrier-degrading proteases during axonal elongation. PMID- 9725907 TI - Selective perturbation of apical membrane traffic by expression of influenza M2, an acid-activated ion channel, in polarized madin-darby canine kidney cells. AB - The function of acidification along the endocytic pathway is not well understood, in part because the perturbants used to modify compartmental pH have global effects and in some cases alter cytoplasmic pH. We have used a new approach to study the effect of pH perturbation on postendocytic traffic in polarized Madin Darby canine kidney (MDCK) cells. Influenza M2 is a small membrane protein that functions as an acid-activated ion channel and can elevate the pH of the trans Golgi network and endosomes. We used recombinant adenoviruses to express the M2 protein of influenza virus in polarized MDCK cells stably transfected with the polymeric immunoglobulin (Ig) receptor. Using indirect immunofluorescence and immunoelectron microscopy, M2 was found to be concentrated at the apical plasma membrane and in subapical vesicles; intracellular M2 colocalized partly with internalized IgA in apical recycling endosomes as well as with the trans-Golgi network marker TGN-38. Expression of M2 slowed the rate of IgA transcytosis across polarized MDCK monolayers. The delay in transport occurred after IgA reached the apical recycling endosome, consistent with the localization of intracellular M2. Apical recycling of IgA was also slowed in the presence of M2, whereas basolateral recycling of transferrin and degradation of IgA were unaffected. By contrast, ammonium chloride affected both apical IgA and basolateral transferrin release. Together, our data suggest that M2 expression selectively perturbs acidification in compartments involved in apical delivery without disrupting other postendocytic transport steps. PMID- 9725908 TI - Organization of highly acetylated chromatin around sites of heterogeneous nuclear RNA accumulation. AB - Histones found within transcriptionally competent and active regions of the genome are highly acetylated. Moreover, these highly acetylated histones have very short half-lives. Thus, both histone acetyltransferases and histone deacetylases must enrich within or near these euchromatic regions of the interphase chromatids. Using an antibody specific for highly acetylated histone H3, we have investigated the organization of transcriptionally active and competent chromatin as well as nuclear histone acetyltransferase and deacetylase activities. We observe an exclusion of highly acetylated chromatin around the periphery of the nucleus and an enrichment near interchromatin granule clusters (IGCs). The highly acetylated chromatin is found in foci that may reflect the organization of highly acetylated chromatin into "chromonema" fibers. Transmission electron microscopy of Indian muntjac fibroblast cell nuclei indicates that the chromatin associated with the periphery of IGCs remains relatively condensed, most commonly found in domains containing chromatin folded beyond 30 nm. Using electron spectroscopic imaging, we demonstrate that IGCs are clusters of ribonucleoprotein particles. The individual granules comprise RNA rich fibrils or globular regions that fold into individual granules. Quantitative analysis of individual granules indicates that they contain variable amounts of RNA estimated between 1.5 and >10 kb. We propose that interchromatin granules are heterogeneous nuclear RNA-containing particles, some of which may be pre-mRNA generated by nearby transcribed chromatin. An intermediary zone between the IGC and surrounding chromatin is described that contains factors with the potential to provide specificity to the localization of sequences near IGCs. PMID- 9725909 TI - Differential expression and functions of cortical myosin IIA and IIB isotypes during meiotic maturation, fertilization, and mitosis in mouse oocytes and embryos. AB - To explore the role of nonmuscle myosin II isoforms during mouse gametogenesis, fertilization, and early development, localization and microinjection studies were performed using monospecific antibodies to myosin IIA and IIB isotypes. Each myosin II antibody recognizes a 205-kDa protein in oocytes, but not mature sperm. Myosin IIA and IIB demonstrate differential expression during meiotic maturation and following fertilization: only the IIA isoform detects metaphase spindles or accumulates in the mitotic cleavage furrow. In the unfertilized oocyte, both myosin isoforms are polarized in the cortex directly overlying the metaphase arrested second meiotic spindle. Cortical polarization is altered after spindle disassembly with Colcemid: the scattered meiotic chromosomes initiate myosin IIA and microfilament assemble in the vicinity of each chromosome mass. During sperm incorporation, both myosin II isotypes concentrate in the second polar body cleavage furrow and the sperm incorporation cone. In functional experiments, the microinjection of myosin IIA antibody disrupts meiotic maturation to metaphase II arrest, probably through depletion of spindle-associated myosin IIA protein and antibody binding to chromosome surfaces. Conversely, the microinjection of myosin IIB antibody blocks microfilament-directed chromosome scattering in Colcemid treated mature oocytes, suggesting a role in mediating chromosome-cortical actomyosin interactions. Neither myosin II antibody, alone or coinjected, blocks second polar body formation, in vitro fertilization, or cytokinesis. Finally, microinjection of a nonphosphorylatable 20-kDa regulatory myosin light chain specifically blocks sperm incorporation cone disassembly and impedes cell cycle progression, suggesting that interference with myosin II phosphorylation influences fertilization. Thus, conventional myosins break cortical symmetry in oocytes by participating in eccentric meiotic spindle positioning, sperm incorporation cone dynamics, and cytokinesis. Although murine sperm do not express myosin II, different myosin II isotypes may have distinct roles during early embryonic development. PMID- 9725911 TI - Human and yeast cdk-activating kinases (CAKs) display distinct substrate specificities. AB - Cell cycle progression is controlled by the sequential functions of cyclin dependent kinases (cdks). Cdk activation requires phosphorylation of a key residue (on sites equivalent to Thr-160 in human cdk2) carried out by the cdk activating kinase (CAK). Human CAK has been identified as a p40(MO15)/cyclin H/MAT1 complex that also functions as part of transcription factor IIH (TFIIH) where it phosphorylates multiple transcriptional components including the C terminal domain (CTD) of the large subunit of RNA polymerase II. In contrast, CAK from budding yeast consists of a single polypeptide (Cak1p), is not a component of TFIIH, and lacks CTD kinase activity. Here we report that Cak1p and p40(MO15) have strikingly different substrate specificities. Cak1p preferentially phosphorylated monomeric cdks, whereas p40(MO15) preferentially phosphorylated cdk/cyclin complexes. Furthermore, p40(MO15) only phosphorylated cdk6 bound to cyclin D3, whereas Cak1p recognized monomeric cdk6 and cdk6 bound to cyclin D1, D2, or D3. We also found that cdk inhibitors, including p21(CIP1), p27(KIP1), p57(KIP2), p16(INK4a), and p18(INK4c), could block phosphorylation by p40(MO15) but not phosphorylation by Cak1p. Our results demonstrate that although both Cak1p and p40(MO15) activate cdks by phosphorylating the same residue, the structural mechanisms underlying the enzyme-substrate recognition differ greatly. Structural and physiological implications of these findings will be discussed. PMID- 9725912 TI - Rho and Rab small G proteins coordinately reorganize stress fibers and focal adhesions in MDCK cells. AB - The Rho subfamily of the Rho small G protein family (Rho) regulates formation of stress fibers and focal adhesions in many types of cultured cells. In moving cells, dynamic and coordinate disassembly and reassembly of stress fibers and focal adhesions are observed, but the precise mechanisms in the regulation of these processes are poorly understood. We previously showed that 12-O tetradecanoylphorbol-13-acetate (TPA) first induced disassembly of stress fibers and focal adhesions followed by their reassembly in MDCK cells. The reassembled stress fibers showed radial-like morphology that was apparently different from the original. We analyzed here the mechanisms of these TPA-induced processes. Rho inactivation and activation were necessary for the TPA-induced disassembly and reassembly, respectively, of stress fibers and focal adhesions. Both inactivation and activation of the Rac subfamily of the Rho family (Rac) inhibited the TPA induced reassembly of stress fibers and focal adhesions but not their TPA-induced disassembly. Moreover, microinjection or transient expression of Rab GDI, a regulator of all the Rab small G protein family members, inhibited the TPA induced reassembly of stress fibers and focal adhesions but not their TPA-induced disassembly, indicating that, furthermore, activation of some Rab family members is necessary for their TPA-induced reassembly. Of the Rab family members, at least Rab5 activation was necessary for the TPA-induced reassembly of stress fibers and focal adhesions. The TPA-induced, small G protein-mediated reorganization of stress fibers and focal adhesions was closely related to the TPA-induced cell motility. These results indicate that the Rho and Rab family members coordinately regulate the TPA-induced reorganization of stress fibers and focal adhesions that may cause cell motility. PMID- 9725910 TI - Activation of androgen receptor function by a novel nuclear protein kinase. AB - Androgen receptor (AR) belongs to the nuclear receptor superfamily and mediates the biological actions of male sex steroids. In this work, we have characterized a novel 130-kDa Ser/Thr protein kinase ANPK that interacts with the zinc finger region of AR in vivo and in vitro. The catalytic kinase domain of ANPK shares considerable sequence similarity with the minibrain gene product, a protein kinase suggested to contribute to learning defects associated with Down syndrome. However, the rest of ANPK sequence, including the AR-interacting interface, exhibits no apparent homology with other proteins. ANPK is a nuclear protein that is widely expressed in mammalian tissues. Its overexpression enhances AR dependent transcription in various cell lines. In addition to the zinc finger region, ligand-binding domain and activation function AF1 of AR are needed, as the activity of AR mutants devoid of these domains was not influenced by ANPK. The receptor protein does not appear to be a substrate for ANPK in vitro, and overexpression of ANPK does not increase the extent of AR phosphorylation in vivo. In view of this, it is likely that ANPK-mediated activation of AR function is exerted through modification of AR-associated proteins, such as coregulatory factors, and/or through stabilization of the receptor protein against degradation. PMID- 9725913 TI - Tim23p contains separate and distinct signals for targeting to mitochondria and insertion into the inner membrane. AB - The Tim23 protein is an essential inner membrane (IM) component of the yeast mitochondrial protein import pathway. Tim23p does not carry an amino-terminal presequence; therefore, the targeting information resides within the mature protein. Tim23p is anchored in the IM via four transmembrane segments and has two positively charged loops facing the matrix. To identify the import signal for Tim23p, we have constructed several altered versions of the Tim23 protein and examined their function and import in yeast cells, as well as their import into isolated mitochondria. We replaced the positively charged amino acids in one or both loops with alanine residues and found that the positive charges are not required for import into mitochondria, but at least one positively charged loop is required for insertion into the IM. Furthermore, we find that the signal to target Tim23p to mitochondria is carried in at least two of the hydrophobic transmembrane segments. Our results suggest that Tim23p contains separate import signals: hydrophobic segments for targeting Tim23p to mitochondria, and positively charged loops for insertion into the IM. We therefore propose that Tim23p is imported into mitochondria in at least two distinct steps. PMID- 9725914 TI - Differential distribution of dynamin isoforms in mammalian cells. AB - Dynamins are 100-kDa GTPases that are essential for clathrin-coated vesicle formation during receptor-mediated endocytosis. To date, three different dynamin genes have been identified, with each gene expressing at least four different alternatively spliced forms. Currently, it is unclear whether these different dynamin gene products perform distinct or redundant cellular functions. Therefore, the focus of this study was to identify additional spliced variants of dynamin from rat tissues and to define the distribution of the dynamin family members in a cultured rat epithelial cell model (Clone 9 cells). After long distance reverse transcription (RT)-PCR of mRNA from different rat tissues, the full-length cDNAs encoding the different dynamin isoforms were sequenced and revealed four additional spliced variants for dynamin I and nine for dynamin III. Thus, in rat tissues there are a total of at least 25 different mRNAs produced from the three dynamin genes. Subsequently, we generated stably transfected Clone 9 cells expressing full-length cDNAs of six different spliced forms tagged with green fluorescent protein. Confocal or fluorescence microscopy of these transfected cells revealed that many of the dynamin proteins associate with distinct membrane compartments, which include clathrin-coated pits at the plasma membrane and the Golgi apparatus, and several undefined vesicle populations. These results indicate that the dynamin family is more extensive than was originally predicted and suggest that the different dynamin proteins are localized to distinct cytoplasmic or membrane compartments. PMID- 9725915 TI - Sequence determinants for regulated degradation of yeast 3-hydroxy-3 methylglutaryl-CoA reductase, an integral endoplasmic reticulum membrane protein. AB - The degradation rate of 3-hydroxy-3-methylglutaryl CoA reductase (HMG-R), a key enzyme of the mevalonate pathway, is regulated through a feedback mechanism by the mevalonate pathway. To discover the intrinsic determinants involved in the regulated degradation of the yeast HMG-R isozyme Hmg2p, we replaced small regions of the Hmg2p transmembrane domain with the corresponding regions from the other, stable yeast HMG-R isozyme Hmg1p. When the first 26 amino acids of Hmg2p were replaced with the same region from Hmg1p, Hmg2p was stabilized. The stability of this mutant was not due to mislocalization, but rather to an inability to be recognized for degradation. When amino acid residues 27-54 of Hmg2p were replaced with those from Hmg1p, the mutant was still degraded, but its degradation rate was poorly regulated. The degradation of this mutant was still dependent on the first 26 amino acid residues and on the function of the HRD genes. These mutants showed altered ubiquitination levels that were well correlated with their degradative phenotypes. Neither determinant was sufficient to impart regulated degradation to Hmg1p. These studies provide evidence that there are sequence determinants in Hmg2p necessary for degradation and optimal regulation, and that independent processes may be involved in Hmg2p degradation and its regulation. PMID- 9725916 TI - Interactions between growth factors and integrins: latent forms of transforming growth factor-beta are ligands for the integrin alphavbeta1. AB - The multipotential cytokine transforming growth factor-beta (TGF-beta) is secreted in a latent form. Latency results from the noncovalent association of TGF-beta with its processed propeptide dimer, called the latency-associated peptide (LAP); the complex of the two proteins is termed the small latent complex. Disulfide bonding between LAP and latent TGF-beta-binding protein (LTBP) produces the most common form of latent TGF-beta, the large latent complex. The extracellular matrix (ECM) modulates the activity of TGF-beta. LTBP and the LAP propeptides of TGF-beta (isoforms 1 and 3), like many ECM proteins, contain the common integrin-binding sequence RGD. To increase our understanding of latent TGF beta function in the ECM, we determined whether latent TGF-beta1 interacts with integrins. A549 cells adhered and spread on plastic coated with LAP, small latent complex, and large latent complex but not on LTBP-coated plastic. Adhesion was blocked by an RGD peptide, and cells were unable to attach to a mutant form of recombinant LAP lacking the RGD sequence. Adhesion was also blocked by mAbs to integrin subunits alphav and beta1. We purified LAP-binding integrins from extracts of A549 cells using LAP bound to Sepharose. alphavbeta1 eluted with EDTA. After purification in the presence of Mn2+, a small amount of alphavbeta5 was also detected. A549 cells migrated equally on fibronectin- and LAP-coated surfaces; migration on LAP was alphavbeta1 dependent. These results establish alphavbeta1 as a LAP-beta1 receptor. Interactions between latent TGF-beta and alphavbeta1 may localize latent TGF-beta to the surface of specific cells and may allow the TGF-beta1 gene product to initiate signals by both TGF-beta receptor and integrin pathways. PMID- 9725918 TI - Tubulin polyglycylation: differential posttranslational modification of dynamic cytoplasmic and stable axonemal microtubules in paramecium. AB - Polyglycylation, a posttranslational modification of tubulin, was discovered in the highly stable axonemal microtubules of Paramecium cilia where it involves the lateral linkage of up to 34 glycine units per tubulin subunit. The observation of this type of posttranslational modification mainly in axonemes raises the question as to its relationship with axonemal organization and with microtubule stability. This led us to investigate the glycylation status of cytoplasmic microtubules that correspond to the dynamic microtubules in Paramecium. Two anti glycylated tubulin monoclonal antibodies (mAbs), TAP 952 and AXO 49, are shown here to exhibit different affinities toward mono- and polyglycylated synthetic tubulin peptides. Using immunoblotting and mass spectrometry, we show that cytoplasmic tubulin is glycylated. In contrast to the highly glycylated axonemal tubulin, which is recognized by the two mAbs, cytoplasmic tubulin reacts exclusively with TAP 952, and the alpha- and beta- tubulin subunits are modified by only 1-5 and 2-9 glycine units, respectively. Our analyses suggest that most of the cytoplasmic tubulin contains side chain lengths of 1 or 2 glycine units distributed on several glycylation sites. The subcellular partition of distinct polyglycylated tubulin isoforms between cytoplasmic and axonemal compartments implies the existence of regulatory mechanisms for glycylation. By following axonemal tubulin immunoreactivity with anti-glycylated tubulin mAbs upon incubation with a Paramecium cellular extract, the presence of a deglycylation enzyme is revealed in the cytoplasm of this organism. These observations establish that polyglycylation is reversible and indicate that, in vivo, an equilibrium between glycylating and deglycylating enzymes might be responsible for the length of the oligoglycine side chains of tubulin. PMID- 9725917 TI - Regulation of the actin cytoskeleton by thrombin in human endothelial cells: role of Rho proteins in endothelial barrier function. AB - Endothelial barrier function is regulated at the cellular level by cytoskeletal dependent anchoring and retracting forces. In the present study we have examined the signal transduction pathways underlying agonist-stimulated reorganization of the actin cytoskeleton in human umbilical vein endothelial cells. Receptor activation by thrombin, or the thrombin receptor (proteinase-activated receptor 1) agonist peptide, leads to an early increase in stress fiber formation followed by cortical actin accumulation and cell rounding. Selective inhibition of thrombin-stimulated signaling systems, including Gi/o (pertussis toxin sensitive), p42/p44, and p38 MAP kinase cascades, Src family kinases, PI-3 kinase, or S6 kinase pathways had no effect on the thrombin response. In contrast, staurosporine and KT5926, an inhibitor of myosin light chain kinase, effectively blocked thrombin-induced cell rounding and retraction. The contribution of Rho to these effects was analyzed by using bacterial toxins that either activate or inhibit the GTPase. Escherichia coli cytotoxic necrotizing factor 1, an activator of Rho, induced the appearance of dense actin cables across cells without perturbing monolayer integrity. Accordingly, lysophosphatidic acid, an activator of Rho-dependent stress fiber formation in fibroblasts, led to reorganization of polymerized actin into stress fibers but failed to induce cell rounding. Inhibition of Rho with Clostridium botulinum exoenzyme C3 fused to the B fragment of diphtheria toxin caused loss of stress fibers with only partial attenuation of thrombin-induced cell rounding. The implication of Rac and Cdc42 was analyzed in transient transfection experiments using either constitutively active (V12) or dominant-interfering (N17) mutants. Expression of RacV12 mimicked the effect of thrombin on cell rounding, and RacN17 blocked the response to thrombin, whereas Cdc42 mutants were without effect. These observations suggest that Rho is involved in the maintenance of endothelial barrier function and Rac participates in cytoskeletal remodeling by thrombin in human umbilical vein endothelial cells. PMID- 9725919 TI - The dynamics of golgi protein traffic visualized in living yeast cells. AB - We describe for the first time the visualization of Golgi membranes in living yeast cells, using green fluorescent protein (GFP) chimeras. Late and early Golgi markers are present in distinct sets of scattered, moving cisternae. The immediate effects of temperature-sensitive mutations on the distribution of these markers give clues to the transport processes occurring. We show that the late Golgi marker GFP-Sft2p and the glycosyltransferases, Anp1p and Mnn1p, disperse into vesicle-like structures within minutes of a temperature shift in sec18, sft1, and sed5 cells, but not in sec14 cells. This is consistent with retrograde vesicular traffic, mediated by the vesicle SNARE Sft1p, to early cisternae containing the target SNARE Sed5p. Strikingly, Sed5p itself moves rapidly to the endoplasmic reticulum (ER) in sec12 cells, implying that it cycles through the ER. Electron microscopy shows that Golgi membranes vesiculate in sec18 cells within 10 min of a temperature shift. These results emphasize the dynamic nature of Golgi cisternae and satisfy the kinetic requirements of a cisternal maturation model in which all resident proteins must undergo retrograde vesicular transport, either within the Golgi complex or from there to the ER, as anterograde cargo advances. PMID- 9725920 TI - Coupled translocation events generate topological heterogeneity at the endoplasmic reticulum membrane. AB - Topogenic determinants that direct protein topology at the endoplasmic reticulum membrane usually function with high fidelity to establish a uniform topological orientation for any given polypeptide. Here we show, however, that through the coupling of sequential translocation events, native topogenic determinants are capable of generating two alternate transmembrane structures at the endoplasmic reticulum membrane. Using defined chimeric and epitope-tagged full-length proteins, we found that topogenic activities of two C-trans (type II) signal anchor sequences, encoded within the seventh and eighth transmembrane (TM) segments of human P-glycoprotein were directly coupled by an inefficient stop transfer (ST) sequence (TM7b) contained within the C-terminus half of TM7. Remarkably, these activities enabled TM7 to achieve both a single- and a double spanning TM topology with nearly equal efficiency. In addition, ST and C-trans signal anchor activities encoded by TM8 were tightly linked to the weak ST activity, and hence topological fate, of TM7b. This interaction enabled TM8 to span the membrane in either a type I or a type II orientation. Pleiotropic structural features contributing to this unusual topogenic behavior included 1) a short, flexible peptide loop connecting TM7a and TM7b, 2) hydrophobic residues within TM7b, and 3) hydrophilic residues between TM7b and TM8. PMID- 9725921 TI - Mutations of the cystic fibrosis gene in patients with chronic pancreatitis. AB - BACKGROUND: The pancreatic lesions of cystic fibrosis develop in utero and closely resemble those of chronic pancreatitis. Therefore, we hypothesized that mutations of the cystic fibrosis transmembrane conductance regulator (CFTR) gene may be more common than expected among patients with chronic pancreatitis. METHODS: We studied 134 consecutive patients with chronic pancreatitis (alcohol related disease in 71, hyperparathyroidism in 2, hypertriglyceridemia in 1, and idiopathic disease in 60). We examined DNA for 22 mutations of the CFTR gene that together account for 95 percent of all mutations in patients with cystic fibrosis in the northwest of England. We also determined the length of the noncoding sequence of thymidines in intron 8, since the shorter the sequence, the lower the proportion of normal CFTR messenger RNA. RESULTS: The 94 male and 40 female patients ranged in age from 16 to 86 years. None had a mutation on both copies of the CFTR gene. Eighteen patients (13.4 percent), including 12 without alcoholism, had a CFTR mutation on one chromosome, as compared with a frequency of 5.3 percent among 600 local unrelated partners of persons with a family history of cystic fibrosis (P<0.001). A total of 10.4 percent of the patients had the 5T allele in intron 8 (14 of 134), which is twice the expected frequency (P=0.008). Four patients were heterozygous for both a CFTR mutation and the 5T allele. Patients with a CFTR mutation were younger than those with no mutations (P=0.03). None had the combination of sinopulmonary disease, high sweat electrolyte concentrations, and low nasal potential-difference values that are diagnostic of cystic fibrosis. CONCLUSIONS: Mutations of the CFTR gene and the 5T genotype are associated with chronic pancreatitis. PMID- 9725922 TI - Relation between mutations of the cystic fibrosis gene and idiopathic pancreatitis. AB - BACKGROUND: It is unknown whether genetic factors predispose patients to idiopathic pancreatitis. In patients with cystic fibrosis, mutations of the cystic fibrosis transmembrane conductance regulator (CFTR) gene typically cause pulmonary and pancreatic insufficiency while rarely causing pancreatitis. We examined whether idiopathic pancreatitis is associated with CFTR mutations in persons who do not have lung disease of cystic fibrosis. METHODS: We studied 27 patients (mean age at diagnosis, 36 years), 22 of whom were female, who had been referred for an evaluation of idiopathic pancreatitis. DNA was tested for 17 CFTR mutations and for the 5T allele in intron 8 of the CFTR gene. The 5T allele reduces the level of functional CFTR and is associated with an inherited form of infertility in males. Patients with two abnormal CFTR alleles were further evaluated for unrecognized cystic fibrosis-related lung disease, and both base line and CFTR-mediated ion transport were measured in the nasal mucosa. RESULTS: Ten patients with idiopathic chronic pancreatitis (37 percent) had at least one abnormal CFTR allele. Eight CFTR mutations were detected (prevalence ratio, 11:1; 95 percent confidence interval, 5 to 23; P<0.001). In three patients both alleles were affected (prevalence ratio, 80:1; 95 percent confidence interval, 17 to 379; P<0.001). These three patients did not have lung disease typical of cystic fibrosis on the basis of sweat testing, spirometry, or base-line nasal potential difference measurements. Nonetheless, each had abnormal nasal cyclic AMP-mediated chloride transport. CONCLUSION: In a group of patients referred for evaluation of idiopathic pancreatitis, there was a strong association between mutations in the CFTR gene and pancreatitis. The abnormal CFTR genotypes in these patients with pancreatitis resemble those associated with male infertility. PMID- 9725923 TI - Spontaneous initiation of atrial fibrillation by ectopic beats originating in the pulmonary veins. AB - BACKGROUND: Atrial fibrillation, the most common sustained cardiac arrhythmia and a major cause of stroke, results from simultaneous reentrant wavelets. Its spontaneous initiation has not been studied. METHODS: We studied 45 patients with frequent episodes of atrial fibrillation (mean [+/-SD] duration, 344+/-326 minutes per 24 hours) refractory to drug therapy. The spontaneous initiation of atrial fibrillation was mapped with the use of multielectrode catheters designed to record the earliest electrical activity preceding the onset of atrial fibrillation and associated atrial ectopic beats. The accuracy of the mapping was confirmed by the abrupt disappearance of triggering atrial ectopic beats after ablation with local radio-frequency energy. RESULTS: A single point of origin of atrial ectopic beats was identified in 29 patients, two points of origin were identified in 9 patients, and three or four points of origin were identified in 7 patients, for a total of 69 ectopic foci. Three foci were in the right atrium, 1 in the posterior left atrium, and 65 (94 percent) in the pulmonary veins (31 in the left superior, 17 in the right superior, 11 in the left inferior, and 6 in the right inferior pulmonary vein). The earliest activation was found to have occurred 2 to 4 cm inside the veins, marked by a local depolarization preceding the atrial ectopic beats on the surface electrocardiogram by 106+/-24 msec. Atrial fibrillation was initiated by a sudden burst of rapid depolarizations (340 per minute). A local depolarization could also be recognized during sinus rhythm and abolished by radiofrequency ablation. During a follow-up period of 8+/-6 months after ablation, 28 patients (62 percent) had no recurrence of atrial fibrillation. CONCLUSIONS: The pulmonary veins are an important source of ectopic beats, initiating frequent paroxysms of atrial fibrillation. These foci respond to treatment with radio-frequency ablation. PMID- 9725924 TI - Risk factors for preeclampsia, abruptio placentae, and adverse neonatal outcomes among women with chronic hypertension. National Institute of Child Health and Human Development Network of Maternal-Fetal Medicine Units. PMID- 9725925 TI - Images in clinical medicine. Thromboangiitis obliterans (Buerger's disease). PMID- 9725926 TI - Use and cost effectiveness of smoking-cessation services under four insurance plans in a health maintenance organization. AB - BACKGROUND: Lack of information about the effect of insurance coverage on the demand for and use of smoking-cessation services has prevented widescale adoption of coverage for such services. METHODS: In a longitudinal, natural experiment, we compared the use and cost effectiveness of three forms of coverage with those of a standard form of coverage for smoking-cessation services that included a behavioral program and nicotine-replacement therapy. The study involved seven employers and a total of 90,005 adult enrollees. The standard plan offered 50 percent coverage of the behavioral program and full coverage of nicotine replacement therapy. The other plans offered 50 percent coverage of both the behavioral program and nicotine-replacement therapy (reduced coverage), full coverage of the behavioral program and 50 percent coverage of nicotine replacement therapy (flipped coverage), or full coverage of both the behavioral program and nicotine-replacement therapy. RESULTS: Estimated annual rates of use of smoking-cessation services ranged from 2.4 percent (among smokers with reduced coverage) to 10 percent (among those with full coverage). Smoking-cessation rates ranged from 28 percent (among users with full coverage) to 38 percent (among those with standard coverage). The estimated percentage of all smokers who would quit smoking per year as a result of using the services ranged from 0.7 percent (with reduced coverage) to 2.8 percent (with full coverage). The average cost to the health plan per user who quit smoking ranged from $797 (with standard coverage) to $1,171 (with full coverage). The annual cost per smoker ranged from $6 (with reduced coverage) to $33 (with full coverage). The annual cost per enrollee ranged from $0.89 (with reduced coverage) to $4.92 (with full coverage). CONCLUSIONS: Use of smoking-cessation services varies according to the extent of coverage, with the highest rates of use among smokers with full coverage. Although the rate of smoking cessation among the benefit users with full coverage was lower than the rates among users with plans requiring copayments, the effect on the overall prevalence of smoking was greater with full coverage than with the cost-sharing plans. PMID- 9725927 TI - Acute vestibular syndrome. PMID- 9725928 TI - Pancreatitis and mutations of the cystic fibrosis gene. PMID- 9725929 TI - Ethical guidelines for physician compensation based on capitation. PMID- 9725930 TI - Capturing the unexpected benefits of medical research. PMID- 9725931 TI - Total and viable airborne bacterial load in two different agricultural environments using gas chromatography-tandem mass spectrometry and culture: a prototype study. AB - Airborne exposure to bacterial components found in agricultural environments can lead to pulmonary inflammation. Total (viable and nonviable) bacterial load was monitored in a stable and a dairy by a new approach, gas chromatography-tandem mass spectrometry measurement of muramic acid, a component of gram positive and gram negative bacterial peptidoglycan. Also used to assess the gram negative bacterial load were 3-hydroxy fatty acids, markers of bacterial lipopolysaccharide. Culture, an established procedure for assessing the viable bacterial portion of airborne dust, served as a basis for comparison. The muramic acid and 3-hydroxy fatty acid concentrations (total C12:0, C14:0, and C16:0) showed a correlation with an R2 of 0.81. Dust and muramic acid levels also correlated. However, although relative muramic acid levels were lower in the stable than the dairy, colony forming units (CFU) were considerably higher in the stable. The total bacterial load (estimated from muramic acid values) for both the stable and dairy was also higher than would have been predicted from culture. These results suggest that nonculture based approaches and culture provide complementary but independent measurements of airborne biopollution. PMID- 9725932 TI - Sampling and analytical method development for qualitative assessment of airborne mycobacterial species of the Mycobacterium tuberculosis complex. AB - This article presents a novel, qualitative approach for detecting airborne M. tuberculosis. Culturing or sample purification is not required. A DNA diagnostic method involving the polymerase chain reaction (PCR) coupled to an enzymatically generated color reaction was used for direct detection of M. bovis BCG (Bacillus of Calmette-Guerin), a surrogate for pathogenic M. tuberculosis. Fewer than 10 mycobacteria were detected with no culturing using this bioanalytical method. Analysis was completed in 1 to 1.5 days, in contrast to traditional culturing methods requiring a minimum of 2-3 weeks. To evaluate an air sampling method coupled to a PCR bioanalytical method, liquid cultures of the surrogate were aerosolized and collected for PCR analyses using 37-mm filter cassettes containing polytetrafluoroethylene filters. An Andersen six-stage (viable) particle sizing sampler was employed as a reference sampler. Aerosolized BCG impacted onto Andersen agar plates required incubation periods of 6-8 weeks before small colony forming units could be detected and enumerated. Although the BCG mean length of the rod-shaped particles was 8.3 microns, the airborne BCG particles were collected predominantly on the Andersen 4-6 stages, representing aerodynamic diameters 0.7 to 3.3 microns. Approximately 25 mycobacteria were detected without culturing using the PCR-filter cassette method. This approach could be used to detect airborne mycobacterial species of the M. tuberculosis complex and could permit the early detection of contaminated indoor air. Also, the efficacy of environmental controls could be evaluated and monitored. This approach could also be used to study the expulsion of infectious particles from patients and may permit risk assessment in regard to personal respiratory protection. PMID- 9725933 TI - ASTM F739 method for testing the permeation resistance of protective clothing materials: critical analysis with proposed changes in procedure and test-cell design. AB - ASTM (American Society for Testing and Materials) Method F739-96 specifies a test cell design and procedures for measuring the permeation resistance of chemical protective clothing. Among the specifications are open-loop collection stream flow rates of 0.050 to 0.150 L/min for a gaseous medium. At elevated temperatures the test must be maintained within 1 degree C of the set point. This article presents a critical analysis of the effect of the collection stream flow rate on the measured permeation rate and on the temperature uniformity within the test cell. Permeation tests were conducted on four polymeric glove materials with 44 solvents at 25 degrees C. Flow rates > 0.5 L/min were necessary to obtain accurate steady-state permeation rate (SSPR) values in 25 percent of the tests. At the lower flow rates the true SSPR typically was underestimated by a factor of two or less, but errors of up to 33-fold were observed. No clear relationship could be established between the need for a higher collection stream flow rate and either the vapor pressure or the permeation rate of the solvent, but test results suggest that poor mixing within the collection chamber was a contributing factor. Temperature gradients between the challenge and collection chambers and between the bottom and the top of the collection chamber increased with the water bath temperature and the collection stream flow rate. Use of a test cell modified to permit deeper submersion reduced the gradients to < or = 0.5 degrees C. It is recommended that all SSPR measurements include verification of the adequacy of the collection stream flow rate. For testing at nonambient temperatures, the modified test cell described here could be used to ensure temperature uniformity throughout the cell. PMID- 9725934 TI - Evaluation of clothing systems to determine heat strain. AB - This article describes the basic evaluation process and test methodology employed when temperature extremes for clothing systems must be considered as part of the U.S. Army's Health Hazard Assessment for material in the development and acquisition process. The goals of the evaluation are to select clothing systems that minimize the hazards of heat strain and to predict the heat strain for persons wearing such clothing. Clothing evaluations begin with biophysical assessments that determine the thermal characteristics (vapor permeability and insulation) for textiles via guarded hot plate tests and for clothing systems via thermal manikin tests. The results from biophysical tests can be used to select the textile and/or clothing with the best thermal characteristics. The data from manikin evaluations also can be used in prediction modeling. Human physiological testing is best done in a controlled laboratory environment, although for realism and user acceptability field trials may also be conducted. Proven test and measurement methods must be employed, and tests must control for confounding variables; subjects serve as their own controls, and test environment and procedures are consistent between trials. The process and test methodology described can be applied to the evaluation of civilian clothing systems as well as to the military systems for which they were developed. PMID- 9725935 TI - A descriptive study of U.S. OSHA penalties and inspection frequency for musculoskeletal disorders in the workplace. AB - Information on the frequency and cost of OSHA enforcement penalties for musculoskeletal disorders (MSD) in the literature is limited. Such information would be of value to organizations in estimating the likelihood and financial impact of enforcement activity in their operations. This descriptive study utilized data from federal Occupational Safety and Health Administration (OSHA) inspections to examine the distribution of penalty costs arising from inspections with MSD-related citations from January 1985 to June 1994 and to estimate the probability of OSHA inspection in general and OSHA citation for MSD hazards from October 1985 to September 1993. The mean and median values of proposed penalties were $47,707 and $3600 respectively. A substantial influence of 1991 changes to the penalty structure was noted with decreasing mean and increasing median penalty values. Penalty values increased with establishment size and were higher for unplanned than for planned inspections. The probability of a federal OSHA inspection for any establishment ranged from 1:50 in 1986 to 1:100 in 1993 whereas the estimated probability of an inspection with MSD-related citations ranged from 1:167,000 in 1996 to as much as 1:38,000 during the peak of enforcement activity in 1990. The probability of an inspection with MSD citations for the largest establishments during the period was more than 1000 times greater than that for the smallest. The results of this study may be utilized by organizations seeking to demonstrate the advantages of reducing musculoskeletal morbidity in the workplace. PMID- 9725936 TI - Laboratory investigation of the mass stability of sampling cassettes from inhalable aerosol samplers. AB - A study was conducted to evaluate the mass stability of the materials used in the construction of samplers with internal cassettes for the gravimetric measurement of inhalable aerosol exposures. The internal cassettes from IOM samplers were studied. Results indicate that the mass stability of filters is uniform, but the mass stability of the cassette material may dramatically affect the results of the measurement. Cassettes constructed from plastic exhibited drastic shifts in mass depending on the environmental conditions of their storage. Under room humidity, the plastic cassettes absorbed 1 to 2 mg of water over several days. When these cassettes were placed in a desiccator, they lost mass consistently but did not approach a stable mass. Studies repeated with cassettes made of stainless steel showed negligible mass variability. Based on this study, the use of stainless steel cassettes is recommended for gravimetric determinations of aerosol exposure, although field blanks may in some cases be used for correction of data from plastic cassettes. This study shows the need to evaluate the mass stability of the cassette material of any sampling device where an internal cassette is weighed together with the filter. PMID- 9725937 TI - Herb remedy. PMID- 9725939 TI - Weightlessness and the human body. PMID- 9725940 TI - Attention-deficit hyperactivity disorder. PMID- 9725941 TI - [New discoveries on target blood pressure and ACE inhibitors. Swedish guidelines on hypertension are still correct]. PMID- 9725943 TI - [Should surgery of inguinal hernia be performed by human "machines"?]. PMID- 9725942 TI - [Nitroglycerin ointment is a mild treatment agent for anal fissure. Surgery may cause permanent injury to the internal anal sphincter]. PMID- 9725944 TI - [Selma Lagerlof was not old at that time]. PMID- 9725945 TI - [A question from Iris: why nothing happens?]. PMID- 9725946 TI - [Who is responsible when seriously ill people travel?]. PMID- 9725947 TI - [Be careful with the -ism!]. PMID- 9725948 TI - [Radiographs of poor quality should have been quality-controlled]. PMID- 9725949 TI - [Incorrect about the prescription rights?]. PMID- 9725950 TI - [Recommendations on the treatment of borrelia infection should be taken with reservation]. PMID- 9725951 TI - [Can Swedish clinical trials keep the world-wide leading position?]. PMID- 9725952 TI - [Aromatherapy a complementary therapy]. PMID- 9725953 TI - [Most of the eye problems can be managed in primary health care. Simple check lists and "languages skills" are often the correct way]. PMID- 9725954 TI - [Left ventricular hypertrophy--an important, often unrecognized risk factor]. AB - Epidemiological studies both in the general population and in series of hypertensive patients have shown the presence of left ventricular hypertrophy (LVH), as determined by echocardiography, to be a powerful risk factor for cardiovascular events. Although LVH is generally associated with hypertension, less than half of hypertensive patients develop LVH. Insulin resistance, endothelial dysfunction, increased blood viscosity, decreased arterial compliance, and increased angiotensin-converting enzyme activity have all been associated with the development of LVH. Although findings in observational studies suggest LVH regression to have a beneficial effect in terms of decreased cardiovascular risk, confirmation of this awaits the results of prospective clinical trials currently being carried out. PMID- 9725955 TI - [Old-age assistance is to be multicultural--a special care for only a minority]. AB - According to the census, more than 100,000 foreign-born people over 65 years of age now live in Sweden. They represent nearly eight per cent of the elderly population, and come from over a hundred different countries. During the 1990s many ethnically and religiously based old-age and health-care services have been developed for this population. The experiences of these services and their importance in the planning of care facilities are discussed in the article; research and scientific evaluations comparing health and social conditions of the foreign-born and Swedish elderly populations are also reviewed. PMID- 9725956 TI - [The number of MRI examinations of cervical spine should be reduced]. PMID- 9725957 TI - [Deficient cooperation between emergency services and primary health care. Can a large gap in the steering system be adequately adjusted?]. PMID- 9725958 TI - [A successful attempt with a consulting service for boys]. AB - As adolescence is a critical period of development, and as boys are less inclined than girls to approach the school facility for adolescent counselling, segregated consulting hours were introduced for boys to attract those with problems. The frequency of consultations by boys increased by 25 per cent, and 70 per cent of the boys reported a preference for the segregated consulting hours; 75 per cent appreciated the absence of girls from the waiting room; and of the 42 per cent with special preferences regarding the gender of the staff encountered, half reported preferring a man. Most of the boys presented with defined problems, though many revealed other problems, often relating to sexuality, in the course of consultation. The availability of segregated consulting hours for boys with adolescent problems is important, and often the only way to reach young boys who need help. PMID- 9725959 TI - [One more case of an Asian fungal infection in an HIV-positive man]. PMID- 9725960 TI - [A survey according to the demands of Psychiatriutredningen in the southern Bohusl[n: How is the living situation of mentally disabled? A study which may be a basis of supportive measures]. PMID- 9725961 TI - [Fibrinolysis or angioplasty in acute myocardial infarction?]. AB - Early reperfusion during myocardial infarction limits myocardial injury and reduces mortality. Fibrinolysis (with streptokinase, or tissue or recombinant plasminogen activators) is today an established method for the treatment of myocardial infarction patients manifesting ST-segment elevation or left bundle branch block at ECG (electrocardiography), effective reperfusion being obtained in fifty per cent of cases. Extensive developments are under way, both of fibrinolytic substances and of various adjuvant treatments. A satisfactory alternative treatment to fibrinolysis is percutaneous transluminal coronary angioplasty (PTCA), a method which can be used when fibrinolysis is contraindicated or during cardiogenic shock, or when there is no sign of reperfusion in response to fibrinolytic treatment. Provided the facilities and competence are available, PTCA can even be used as primary treatment instead of fibrinolysis. PMID- 9725963 TI - [Only shadows remain in Spinalonga]. PMID- 9725962 TI - [Defective iron metabolism in genetic hemochromatosis. The mechanisms remain unknown in spite of genetic advances]. AB - Genetic haemochromatosis (GH) is one of the most common hereditary diseases, with a prevalence of 1-5/1000 in the Western world. In 90 per cent of cases a mutation is found in an MHC-class-like gene designated HFE, involving a substitution at position 282 of the HFE protein and resulting in defective binding of beta(2) microglobulin. Animals with beta(2)-microglobulin deficiency develop iron overload, indicating this protein to be involved in the regulation of iron metabolism. Hepatic iron overload results in increased production of oxygen free radicals and peroxidation of membrane lipids, thus causing damage to lysosomes, mitochondria and the endoplasmic reticulum. These cellular events may progress to cell death, fibrogenesis, and the development of liver cirrhosis which is associated with a 200-fold increase in risk of hepatocellular carcinoma. In addition to the risk of diabetes, arthralgia, cardiac arrhythmia, pituitary insufficiency and hypogonadism, iron excess is also associated with aggravation of the cytotoxic effects exerted on hepatocytes by other agents such as alcohol or hepatotrophic viruses. The treatment of iron overload in GH consists of weekly venesection until the serum ferritin level is normalized, followed by maintenance therapy. Survival rates are normal if the disease is detected and treated before complications have developed. PMID- 9725964 TI - 1997 annual report of the American Association of Poison Control Centers Toxic Exposure Surveillance System. PMID- 9725965 TI - Effects of risperidone in overdose. AB - This study was a 13-month prospective, descriptive case series of risperidone overdose reported by telephone to a regional poison control center (PCC) serving Philadelphia, PA. Patients were seen in local Philadelphia-area emergency departments. The variables examined were medical history, therapeutic use of risperidone, time postingestion, reported coingestants, clinical findings, decontamination and treatment, electrocardiograph results, laboratory data, standard toxicologic screen results, and length of time in hospital. Thirty-one patients (29 adult/adolescent, 2 pediatric) with reported risperidone overdose were identified. Risperidone was the sole ingestant in 15 cases (1 mg to 180 mg). The major observed effects in this group included lethargy (7), spasm/dystonia (3), hypotension (2), tachycardia (6), and dysrhythmia (1). Sixteen cases involved coingestants, including benzodiazepines, selective serotonin reuptake inhibitors, ethanol, tricyclic antidepressants, lithium, anticonvulsants, diphenhydramine, ibuprofen, and anticholinergic agents. Major effects in these patients included lethargy (10), coma (1), seizure (1), tachycardia (7), bradycardia (1), hypotension (4), and a syndrome of muscle spasms, diaphoresis, and fever. Treatment provided for patients in this study included antiarrhythmics (1), diphenhydramine (2), anticonvulsant (1), vasopressor agent (1), endotracheal intubation/assisted ventilation (5), and supportive care. One patient who coingested imipramine died of medical complications. In the remaining patients, symptoms resolved with 24 hours in the majority, with all patients asymptomatic at 72 hours postingestion. These data show that risperidone toxicity manifests primarily as mild central nervous system effects and reversible neuromuscular and cardiovascular effects. PMID- 9725966 TI - Postpartum myocardial infarction induced by methergine. AB - A 28-year-old woman presented to the emergency department for evaluation of acute chest pain. She lacked risk factors for coronary artery disease and her initial electrocardiogram (ECG) was nondiagnostic. Within 45 minutes of presentation she developed nausea, vomiting, restrosternal chest pain, and ECG changes compatible with an acute inferoposterior myocardial infarction. Emergent cardiac catheterization revealed three-vessel coronary artery ectasia and two-vessel occlusion. She underwent emergency coronary artery bypass grafting. Her myocardial ischemia was believed to have been induced by methergine, which she had been taking over the preceding 3 days. The etiology and pathophysiology of coronary artery ectasia, as well as the cardiovascular effects of methergine and a related drug, ergotamine, are discussed. PMID- 9725967 TI - Treatment of acute anticholinergic poisoning with physostigmine. AB - Five cases of acute anticholinergic poisoning presenting to an inner-city emergency department (ED) are discussed. All five patients presented with classic signs and symptoms of anticholinergic toxicity, which included tachycardia, hot, dry and flushed skin, markedly dilated and fixed pupils, and pronounced delirium. The patients were violently agitated, and physical restraint was required. Initial treatment with benzodiazepines did not diminish their combative behavior. Treatment with intravenous physostigmine salicylate resulted in a decrease in agitation within 15 to 20 minutes of therapy. No untoward effects occurred as a result of treatment with physostigmine. PMID- 9725968 TI - Skin necrosis and venous thrombosis from subcutaneous injection of charcoal lighter fluid (naptha). AB - A 33-year-old white man injected approximately 4 cc of charcoal lighter fluid (99.4% naptha/0.6% inert ingredients) subcutaneously into his left antecubital fossa. The injection resulted in the toxic necrosis of his volar forearm skin extending proximally to mid-humerus and distally to the metacarpophalangeal joints of the left hand dorsally over a 6-day period. The patient ultimately required extensive surgical debridement, secondary operative closure, and approximately 150 cm2 of split-thickness skin grafting. This case demonstrates the potential for widespread, delayed toxic necrosis of the skin resulting from subcutaneous injection of naptha. This patient's case appears to represent the most severe and widespread case of toxic necrosis of the skin resulting from the subcutaneous injection of hydrocarbons reported in the literature. This case also demonstrates extensive toxic thrombophlebitis not reported in prior cases involving subcutaneous injection of hydrocarbons. PMID- 9725969 TI - Anaphylaxis to topical bacitracin zinc ointment. AB - This report describes a case of systemic anaphylaxis to bacitracin zinc ointment in a 24-year-old man who was injured in a motorcycle accident. Extensive abrasions on the patient's extremities were cleaned with Shurclens before application of viscous Xylocaine and bacitracin zinc ointment. Five minutes later, the patient exhibited symptoms of severe anaphylaxis and required the administration of epinephrine, antihistamines, intravenous fluids, and corticosteroids. Two weeks later, he underwent prick/puncture skin testing to Shurclens and bacitracin zinc ointment as well as prick/puncture, intracutaneous, and subcutaneous challenge with Xylocaine. Only the result of the prick test to bacitracin zinc ointment was positive. Although bacitracin is considered to be a safe topical antibiotic, physicians should be aware of the potential not only for delayed hypersensitivity but also for acute IgE-mediated allergic reactions and life-threatening anaphylaxis. PMID- 9725970 TI - Airway edema following household bleach ingestion. AB - Household bleach is a frequent nonpharmaceutical childhood ingestion in the US. It is regarded as a benign ingestion. A rare presentation is reported of poisoning by low-concentration hypochlorite household bleach in a toddler that led to severe respiratory sequelae. The literature on respiratory and nonrespiratory sequelae of liquid household bleach ingestion is reviewed. PMID- 9725971 TI - Anticholinergic poisoning with adulterated intranasal cocaine. AB - In recent years, emergency physicians have encountered a growing number of patients who present with anticholinergic toxicity after using adulterated heroin. Anticholinergic poisoning caused by adulterated cocaine is far less common. This report describes the case of a 39-year-old man who arrived in the emergency department several hours after the nasal insufflation of cocaine. Classic symptoms of anticholinergic toxicity were evident on examination, including dry, flushed skin, agitation, tachycardia, mydriasis, and absence of bowel sounds. Treatment included intravenous fluids and lorazepam, with resolution of symptoms over several hours. Urine samples revealed the presence of cocaine metabolites as well as the anticholinergic drug atropine, and infrequently encountered adulterant of cocaine. Anticholinergic poisoning is reviewed, and the physical examination findings that distinguish this syndrome from the closely related sympathomimetic syndrome typical of cocaine are detailed. Current treatment recommendations for anticholinergic poisoning are summarized. PMID- 9725972 TI - Systemic and topical effects of intradermal hydrofluoric acid. AB - A 35-year-old man attempted suicide by subcutaneous self-administration of hydrofluoric acid (5 cc of a domestic rust removal solution containing 7% hydrofluoric acid). A burn 9 x 7 cm in diameter immediately appeared at the injection site (left forearm and fold of the elbow). In the emergency department, the burn was copiously washed with isotonic solution and treated with cutaneous and subcutaneous injections of magnesium chloride, 10% solution of calcium gluconate, and 2% xylocaine, then continuously maintained under topical treatment with calcium gluconate. Seven hours after injection, the patient was severely hypocalcemic (Ca+2 0.64 mmol/L). Ten hours after injection, in addition to the persistent hypocalcemia (Ca+2 0.81 mmol/L), hyponatremia (123 mmol/d), hypokalemia (3.4 mmol/L), and hypochloremia (95.6 mmol/L) had developed. The hypocalcemia was corrected with infusion of calcium gluconate (8.92 mEq of Ca+2 as total amount). The patient underwent surgical intervention 7 days after admission, followed by several interventions of plastic surgery. PMID- 9725973 TI - Acute pancreatitis following lisinopril rechallenge. AB - Acute pancreatitis has many causes, the most common being biliary tract disease and alcoholism. Other etiologic categories are abdominal trauma; postoperative, including endoscopic retrograde cholangiopancreatography; metabolic, including hypercalcemia and hypertriglyceridemia; Infectious; idiopathic; and drug-induced. The drugs most strongly associated with pancreatitis are sulfonamides, thiazides, furosemide, estrogens, and tetracycline. Approximately 100 cases of pancreatitis induced by angiotensin-converting enzyme inhibitor have been reported to the US Food and Drug Administration, of which about 20 involved lisinopril. We report a case of pancreatitis occurring only 3 hours after intake of lisinopril by a man without other risk factors for the illness. The patient had experienced a similar but less severe reaction to this medication 3 months earlier. This case probably represents the first time a patient was rechallenged with lisinopril and had a more significant adverse reaction. PMID- 9725974 TI - Periorbital urticaria and topical fluorescein. AB - Topical fluorescein application is a routine component of the ophthalmic examination. Although complications of fluorescein angiography have been well documented, adverse reactions to topical application of this agent have not been previously reported. This report describes a case of topical fluorescein-induced urticaria. PMID- 9725975 TI - Amlodipine overdose causes prolonged calcium channel blocker toxicity. AB - Amlodipine is a relatively new agent that has the longest half-life of all calcium channel blockers. This report describes a severe overdose that resulted in prolonged and severe hemodynamic compromise for up to 10 days, but responded to aggressive therapy with calcium, glucagon, and other vasoactive medicines. PMID- 9725976 TI - Multiple cocaine-induced seizures and corresponding cocaine and metabolite concentrations. AB - The etiology of seizures associated with cocaine use is unclear. Because cocaine seizures are relatively uncommon, they should be diagnosed by exclusion and a neurological workup to rule out central nervous system (CNS) catastrophe should be made. This report describes the clinical findings, treatment, and blood cocaine and metabolite concentrations in a patient who, on two separate occasions, had seizures associated with crack cocaine ingestion. Approximately 1 hour after the ingestion incidents, the patient had multiple, generalized seizures that abated spontaneously. His workup for CNS bleeding, infection, and trauma was negative. Cocaine concentrations on the first incident peaked at 2.48 mg/L and on the second incident peaked at 3.9 mg/L. Other clinical findings included tachycardia, hypertension, diaphoresis, and disorientation. Blood cocaine and metabolite analysis revealed extremely high concentrations. Other than the incident of seizures and transient cardiovascular aberrations, these high concentrations were tolerated by the patient without further sequelae. A review of cocaine-induced seizures and treatment is included. PMID- 9725977 TI - Hypertonic sodium bicarbonate in an acute flecainide overdose. PMID- 9725978 TI - Use of pharmacokinetics to determine the duration of dialysis in management of methanol poisoning. PMID- 9725979 TI - Warfarin-induced skin necrosis. AB - Skin necrosis is an uncommon complication of warfarin (Coumadin; Dupont Pharma, Wilmington, DE) therapy. The presentation may mimic other disorders. This article reports a case of a 72-year-old woman who presented to the emergency department complaining of swelling and ecchymosis to her left breast and right foot. The patient had been hospitalized for coronary artery bypass grafting, and had been discharged from the hospital earlier that day. This article reviews the pathophysiology and clinical features of warfarin-induced skin necrosis. PMID- 9725980 TI - Alcoholic beverages: proof and flammability. PMID- 9725981 TI - Acute gastric perforation in a chloral hydrate overdose. PMID- 9725982 TI - Trinder's bedside test for qualitative determination of salicylate ingestions. PMID- 9725983 TI - Spontaneous pneumothorax in cocaine sniffers. PMID- 9725984 TI - Ibuprofen overdose presenting with severe agitation and hypothermia. PMID- 9725986 TI - 11th Symposium on Molecular Biology of Hematopoiesis and Treatment of Myeloproliferative Diseases. Bormio, Italy, June 25-29, 1998. Abstracts. PMID- 9725985 TI - Acute hepatitis induced by omeprazole. PMID- 9725987 TI - Clair Patterson's influence on environmental research. PMID- 9725988 TI - Modulation of O6-alkylating agent induced clastogenicity by enhanced DNA repair capacity of bone marrow cells. AB - The murine bone marrow micronucleus assay has been used to examine (1) the potentiation of fotemustine and streptozotocin induced-clastogenicity by the O6 alkylguanine-DNA alkyltransferase (ATase) inactivator O6-benzylguanine (O6-beG) and (2) the level of protection afforded against this potentiation by retrovirus mediated expression of an O6-beG-resistant mutant of human ATase (haTPA/GA) in mouse bone marrow. Both fotemustine and streptozotocin induced significantly higher levels of micronucleated polychromatic erythrocytes (p < 0.001 for the highest doses studied) compared to those seen in vehicle-treated animals. The number of micronuclei produced by either agent was dramatically elevated by pretreatment with O6-beG (p < 0.001). Furthermore, in myeloablated mice reconstituted with bone marrow expressing the O6-beG-resistant hATPA/GA as a result of retroviral gene transfer, the frequency of micronucleus formation following exposure of mice to otherwise clastogenic doses of fotemustine or streptozotocin, in the presence of O6-beG, wash highly significantly reduced (p < 0.001 for both agents) relative to that in mock transduced controls. These data clearly implicate O6-chloroethyl- and O6-methylguanine as clastogenic lesions in vivo and establish ATase as a major protective mechanism operating to reduce the frequency of such damage. The potentiation of drug induced clastogenicity by O6 beG suggests that the clinical use of this inactivator in combination with O6 alkylating agents, could substantially increase the risk of therapy related malignancy. Nevertheless the use of hATPA/GA as a protective mechanism via gene therapy may overcome this risk. PMID- 9725989 TI - Antimutagenic activity of carotenoids in green peppers against some nitroarenes. AB - In Mexico, as well as in Central and South American countries, the consumption of peppers (Capsicum annuum) has been tradition for thousands of years; the per capita dietary intake of peppers is about 40 g/day. Peppers are an important source of beta-carotene and vitamin A, which have antimutagenic and/or anticarcinogenic properties. In the present study, Salmonella typhimurium tester strain YG1024 in the plate-incorporation test was used to examine the antimutagenicity of carotenois extracted from five different types of Capsicum spp. ('Chilaca', 'Poblano', 'Serrano', 'Jalapeno' and 'Pimiento') which were chosen, based on their consumption and availability on the local market. Extracts from these peppers were tested against 1-6-dinitropyrene (1,6-DNP) and 1,8 dinitropyrene (1,8-DNP) mutagenicity. Dose-response mutagenicity curves of 1-NP; 1,6-DNP and 1,8-DNP were obtained. For the antimutagenicity studies, doses of 0.05 microgram/plate, 0.20 ng/plate and 0.06 ng/plate for 1-NP, 1,6-DPN and 1,8 DNP respectively were chosen, and the number of net revertants/plate were 1008 for 1-NP, 512 for 1,6-DNP, and 712 for 1,8-DPN. Trans-beta-carotene and the extracts were not toxic to the bacteria at the concentrations tested. The extracts obtained from the peppers showed more inhibition than pure trans-beta carotene on 1-NP; 1,6-DNP and 1,8-DNP mutagenicity. Chilaca pepper extract required 0.36 g (34 nmol expressed as trans-beta-carotene equivalents) of fresh pepper to inhibit 94% on 1-NP mutagenicity, 78% on 1,6-DNP mutagenicity and 84% on 1,8-DNP mutagenicity. Bell pepper ('Pimiento') extract required 1.53 g (50 nmol expressed as trans-beta-carotene) to obtain 87%, 79% and 73% inhibition on 1 NP; 1,6-DNP and 1,8-DNP mutagenicity respectively. Since pure beta-carotene inhibited only approximately 50% the mutagenicity of nitroarenes, these results suggest that each one of the pepper extracts have more than one antimutagenic compound (e.g., beta-carotene and xanthophylls) and those functional nutrients apparently have a synergistic effect. PMID- 9725990 TI - Application of the comet assay for monitoring DNA damage in workers exposed to chronic low-dose irradiation. I. Strand breakage. AB - We examined a group of people professionally at risk of exposure to low doses of ionizing radiation (altogether 49 individuals). Age, use of therapeutic drugs, work-related exposure to hazardous agents, previous exposures to diagnostic X rays, such as patient and nuclear medical examination, were registered. For each individual, the occupational radiation burden received over the past period of 5 years was taken from the official personal records based on film dosimetry controlled every month. A matched group of controls was chosen among the administrative employees (40 individuals). The mean age of the studied population at the time of blood sampling was 49 years (range 24-69). The individuals were divided into groups according to risk of exposure and sex. The alkaline comet assay was used to measure DNA breaks and alkali-labile sites. We compared the mean tail moments, tail length and percentage of DNA in the tail. There was a significant difference between the control and hazard groups in DNA damage. Higher DNA damage was also found for men than for women in the control group. There was no relation of DNA damage to age either in control or hazard group. Additionally, analysis of distributions of tail moment values pointed to a considerable individual diversity even in the control group. Therefore, further investigations were necessary into the suitability of the comet assay as a biological dosimetry method; the results obtained so far warrant such investigations. PMID- 9725991 TI - Application of the comet assay for monitoring DNA damage in workers exposed to chronic low-dose irradiation. II. Base damage. AB - In the preceding paper [M. Wojewodzka, M. Kruszewski, T. Iwanenko, A.R. Collins, I. Szumiel, Application of the comet assay for monitoring DNA damage in workers exposed to chronic low dose irradiation. I. Strand breakage., Mutat. Res. 416 (1998) 21-35], we reported the results of DNA damage examination carried out for a group of people (49 individuals) professionally at risk of exposure to low doses of ionizing radiation as measured by the alkaline comet assay. Here, we used the method in combination with oxidative base damage-specific endonucleases to estimate base damage in the same individuals. These were endonuclease III (endoIII) and formamidopyrimidine glycosylase (FPG). In contrast to the previous investigations, we found no statistically significant difference in base damage between the control and hazard groups. Interestingly, the hazard group exhibited lower level of enzyme-sensitive sites than the control; however, this different was not significant. No correlation of base damage with age was found, similarly as in the case of DNA damage measured by the alkaline comet assay. Interindividual variability of base damage precluded exposure estimation for single individuals, since several members of the control group exhibited high comet parameters. PMID- 9725992 TI - Cytogenetic consequences after occupational exposure to antineoplastic drugs. AB - Cytogenetic monitoring was carried out on a group of 38 nurses who reconstitute antineoplastic drugs in order to determine the extent of chromosomal damage. Genotoxic activities of antineoplastic drugs are studied by chromosome aberration assay, micronucleus assay, sister chromatid exchange (SCE) frequency high frequency cells (HFC) analysis, and mitotic activity of peripheral lymphocytes. Results confirmed that occupational exposure to a mixture of antineoplastic drugs may cause genome damages. The results of this study show that biomonitoring after exposure to a mixture of antineoplastic drugs which express clastogenic and aneugenic activity should involve a battery of cytogenetic methods. PMID- 9725993 TI - Coke oven workers study: the effect of exposure and GSTM1 and NAT2 genotypes on DNA adduct levels in white blood cells and lymphocytes as determined by 32P postlabelling. AB - The DNA adduct levels in total white blood cells (WBC) and lymphocytes (LYM) isolated from the blood of the same individuals were evaluated using the 32P postlabelling assay for bulky aromatic adducts. In this study, 68 male coke oven workers and 56 machines workers as a matched control were enrolled. Personal monitors were used to evaluate exposure to eight carcinogenic PAHs, including B[alpha]P, during an 8-h working shift. The exposure among coke even workers ranged widely from 0.6 to 547 micrograms/m3 and from 2 to 62,107 ng/m3, for carcinogenic PAHs and B[alpha]P, respectively. The respective values in controls were from 0.07-1.64 microgram/m3 and from 1-63 ng/m3. A significant correlation between WBC- and LYM-DNA adduct levels was found (r = 0.591, P < 0.001). DNA adduct levels in both WBC and LYM were significantly elevated in coke oven workers as compared with controls, but adduct levels were generally low (WBC: medians 2.61 vs. 1.83 LYM: 2.47 vs. 1.65 adducts/10(8) nucleotides). LYM-DNA adduct levels were significantly higher for smokers as compared with nonsmokers in both the exposed and control groups. No such differences in WBC-DNA adduct levels were observed. Positive significant correlations were found at the individual level between DNA adducts in both cell types and carcinogenic PAHs and/or B[alpha]P in the inhaled air (r = 0.38-0.45, P < 0.001). A significant correlation at the individual level between LYM-DNA adducts and urinary cotinine was also observed (r = 0.37, P < 0.001). No differences in DNA adduct levels could be attributed to GSTM1 or NAT2 genotype in either group. Nor was there any clear association of DNA adduct levels with combined GSTM1/NAT2 genotypes. The effect of personal exposure to carcinogenic PAHs on DNA adduct levels in both cell types was also investigated using a logistic regression model with adjustment for possible modulating effect of confounders (smoking, GSTM1, NAT2, age, plasma levels of vitamins A and E, body mass index and diet). The results showed that coke oven workers had a significantly (P < 0.05) increased adjusted Odds Ratio (OR = 4.2 and 3.9 for WBC and LYM-DNA adducts) for occurrence of higher DNA adduct levels as compared to controls. The results also showed that the relative risk of an increased prevalence of 'abnormal' values of DNA adduct levels was exposure-dose related. The influence of confounding variables was found not to be significant in this study of relatively limited size. In spite of this, the results suggest that the DNA adduct levels in LYM seem to be affected by smoking (OR = 1.8 for smokers) and are modulated by the influence of NAT2 genotypes (OR = 1.6 for slow acetylators). Our findings indicate that both cell types are generally suitable to monitor occupational exposure to PAHs, and the results suggest that coke oven workers, smoking individuals and slow acetylators sustain more genetic damage in their LYM-DNA from exposure to carcinogenic PAHs than individuals without these actors. PMID- 9725994 TI - The effect of flavonoids on ofloxacin-induced mutagenicity in Euglena gracilis. AB - The antimutagenicity of 14 naturally occurring flavonoids (20 mumol/l) on ofloxacin (43 mumol/l and 86 mumol/l)-induced bleaching (mutagenicity) was studied in Euglena gracilis. The flavonoids chrysin, techtochrysin, chrysin-5 methylether galangin, galangin-5-methylether, pinocembrin and pinobanksin possess considerable antimutagenic properties against ofloxacin-induced bleaching of E. gracilis. Apigenin and isalpinin had only weak antimutagenic potency. Pinobanksin 5-methylether and pinobanksin-3-acetate showed very weak or no antimutagenic effect. However, kempferol, quercetin-3-methylether and quercetin-3,3' dimethylether showed co-mutagenic or no antimutagenic effect depending on the concentration of ofloxacin. Two possible modes of action of the flavonoids on ofloxacin-induced bleaching of E. gracilis are discussed. PMID- 9725995 TI - Lack of genotoxicity of the herbicide atrazine in cultured human lymphocytes. AB - The widely used herbicide atrazine was evaluated for genotoxicity in cultured human peripheral blood lymphocytes. Sister-chromatid exchanges (SCE), chromosome aberrations (CA) and micronuclei (MN) were scored as genetic endpoints. To detect eventual metabolic modification in the genotoxicity of this herbicide, the cultures of SCE and MN demonstration were also treated with S9 microsomal fraction. From our results we can conclude that atrazine was able to exert a weak cytotoxic effect. However, the overall evaluation of the genotoxicity data indicate that this herbicide is not effective in the three assays conducted, irrespective of the presence of metabolic activation, which would mean a general lack of effectiveness of atrazine to induce clastogenic and aneugenic damage in cultured human lymphocytes. PMID- 9725996 TI - Cytogenetic analysis of South Asian berry pickers in British Columbia using the micronucleus assay in peripheral lymphocytes. AB - Micronuclei in peripheral blood lymphocytes from British Columbia seasonal farmworkers and controls were evaluated using the cytokinesis-block technique. The farmworkers harvested berry crops and were likely occupationally exposed to pesticides. Subjects were 39 female subjects of South Asian descent; 18 farmworkers employed during 1993 and 21 age-matched controls. The mean age was 55.9 years. Micronuceli were also scored for the presence of kinetochores. No significant difference was found between the frequency of micronucleated binucleates in the farmworkers group (19.20/1000 binucleates), and the control group (21.76/1000 binucleates). However, among the farmworkers employed in 1993, there was a positive, but not statistically significant, association between micronucleated cell frequency and weeks worked: 16.44/1000 binucleates in those working less than 20 weeks; 23.78/1000 binucleates in those working 20 to 23 weeks; and 25.43/1000 binucleates in those working more than 23 weeks. In those who had ever been employed as farmworkers, there was an elevated frequency of micronucleated cells in the group with the longest history of employment as a farmworker (25.28/1000 binucleates) compared to those with the shortest employment history (16.48/1000 binucleates). This trend remained evident after adjusting for age, red blood cell folate, meat consumption, coffee consumption and recent vaccination. A positive association between the consumption of meat and micronucleus frequency was also observed. Non-meat eaters were likely life long vegetarians. Micronuclei in farmworkers had a lower frequency of kinetochore positive micronuclei than controls. This study indicates that South Asian berry pickers in British Columbia may be at risk for genetic damage. More studies in other ethnic groups and in males are needed to generalize the findings of this study. More direct measures of exposure are needed to elucidate the sources of genotoxicity. PMID- 9725997 TI - Studies on the genotoxicity of the mammalian lignans enterolactone and enterodiol and their metabolic precursors at various endpoints in vitro. AB - The mammalian lignans enterolactone (ENL) and enterodiol (END) are formed by intestinal bacteria from the plant lignans matairesinol (MAT) and secoisolariciersinol (SEC), respectively, which are ingested with different types of food. ENL and END are weak estrogens. According to epidemiological and biochemical studies, lignans may act as anticarcinogens, but little is known about their genotoxic potential. We have therefore investigated the effects of ENL, END, MAT and SEC on cell-free microtubule assembly and at the following genetic endpoints in cultured male Chinese hamster V79 cells: disruption of the cytoplasmic microtubule complex, induction of mitotic arrest, induction of micronuclei and their characterization by CREST staining, and mutagenicity at the HPRT gene locus. The lignans were tested at concentrations of 200 microM in the cell-free system and 100 microM in cultured cells, which represents the limit of solubility in each assay. The established aneuploidogen diethylstilbestrol and the clastogen 4-nitroquinoline-N-oxide were used as positive reference compounds. As none of the four lignans had any activity at the endpoints studied, we conclude that ENL, END, MAT and SEC are devoid of aneuploidogenic and clastogenic potential under the experimental conditions used in this study. PMID- 9725999 TI - Mutagenicity testing (+/-)-camphor, 1,8-cineole, citral, citronellal, (-)-menthol and terpineol with the Salmonella/microsome assay. AB - The essential oils and their monoterpenoid constituents have been widely used as fragrances in cosmetics, as flavouring food additives, as scenting agents in a variety of household products, as active ingredients in some old drugs, and as intermediates in the synthesis of perfume chemicals. The present study was undertaken to investigate the mutagenic potential of six monoterpenoid compounds: two aldehydes (citral and citronellal), a ketone ((+/-)-camphor), an oxide (1,8 cineole, also known as eucalyptol), and two alcohols (terpineol and (-)-menthol). It is part of a more comprehensive toxicological screening of monoterpenes under way at our laboratory. Mutagenicity was evaluated by the Salmonella/microsome assay (TA97a, TA98, TA100 and TA102 tester strains), without and with addition of an extrinsic metabolic activation system (lyophilized rat liver S9 fraction induced by Aroclor 1254). In all cases, the upper limit of the dose interval tested was either the highest non-toxic dose or the lowest dose of the monoterpene toxic to TA100 strain in the preliminary toxicity test. No mutagenic effect was found with (+/-) camphor, citral, citronellal, 1,8-cineole, and (-) menthol. Terpineol caused a slight but dose-related increase in the number of his+ revertants with TA102 tester strain both without and with addition of S9 mixture. The results from this study therefore suggest that, with the exception of terpineol, the monoterpenoid compounds tested are not mutagenic in the Ames test. PMID- 9725998 TI - Inhibition of PhIP mutagenicity by caffeine, lycopene, daidzein, and genistein. AB - The heterocyclic amine 2-amino-1-methyl-6-phenylimidazo[4,5-beta]pyridine NPhIP) is a major dietary component in individuals eating cooked meats or fish. This heterocyclic amine requires biochemical activation, mainly through cytochrome P4501A2, and can be detoxified chiefly by 4'hydroxylation through other cytochromes, and be in turn converted through phase 2 enzymes to readily excreted conjugates. The active form of PhIP is mutagenic in Salmonella typhimurium TA98 and is a useful substrate to study the possible chemoprotective action of phytochemicals. We found that black and green tea depressed the mutagenicity of PhIP in dose-related fashion, and decaffeinated tea was less powerful an inhibitor. This led to the study of caffeine, that displayed effective dose related inhibition of the mutagenicity of PhIP. Other antioxidants such as lycopene, the active antioxidant from tomatoes, and daidzein and genistein from soy products, also had a dose-related inhibition of the mutagenicity of PhIP. We conclude that PhIP is a good substrate found in several human foods to determine the protective effect of phytochemicals from vegetables, and beverages. PMID- 9726000 TI - Investigation of lung tumour induction in C3H/HeH mice, with and without tumour promotion with urethane, following paternal X-irradiation. AB - In series of papers Nomura has reported that parental irradiation can lead to an enhanced incidence of lung and other tumours. However, in a recent study with BALB/cJ mice, using optimum conditions as defined by Nomura, we were unable to confirm this. We have now repeated the investigation using a different inbred strain, C3H/HeH, with and without tumour promotion in the F1 by urethane, again using protocols defined by Nomura. In a series of replicate studies spanning over 2 years, males were exposed to single, acute doses of 0, 250 and 500 cGy X-rays and thereafter placed with two females each in each of two consecutive weeks. Half the offspring from each treatment group and each week of mating were given 5 mmol/kg body weight of the urethane, while the remainder remained untreated. Most of the offspring produced were killed and scored for lung tumours at 6 months of age, while the rest were examined at 12 months of age. The proportion of fertile females and litter size provided evidence of a dose-dependent mutational response to the paternal irradiation, but no trace of a radiation-enhanced lung tumour incidence was detected among the progeny, whether in the urethane or non-urethane groups at 6 or 12 months of age, and whether assessed by numbers of mice with tumours, clusters of tumours, or cluster size. As seen in the BALB/cJ study, significant differences among different replicates were found, again suggesting a cyclical or seasonal variation in tumour incidence, but the variations seen with the two strains were not the same. The need for concurrent controls for tumour work was, nevertheless, again indicated. The overall findings do not therefore accord with those of Nomura. Furthermore, they do not support the causal association between the raised incidence of childhood leukaemia and non-Hodgkins lymphoma near Sellafield and the father's recorded radiation exposure during employment in the nuclear industry, as suggested by the Gardner report. PMID- 9726001 TI - Genotoxicity and oxidative stress induced by pesticide exposure in bovine lymphocyte cultures in vitro. AB - The genotoxic activity of the pesticides gliphosate, vinclozolin and DPX-E9636 was studied in in vitro cultures of bovine lymphocytes, using chromosome aberration (CA) and sister chromatid exchange (SCE) frequencies as genetic end points and a variation of glucose 6-phosphate dehydrogenase (G6PD) enzyme activity as a marker of changes in the normal cell redox state. Results indicated a statistically significant increase of structural aberrations, sister chromatid exchanges and G6PD activity, suggesting that the pesticides tested induce either oxidative stress or a mutagenic effect in this species. The evaluation of both mitotic index and cell viability, after pesticide exposure, demonstrates a high cytotoxic effect which is always associated with the observed genotoxic effect. PMID- 9726003 TI - Nitro-polycyclic aromatic hydrocarbon contents of fumes from heated cooking oils and prevention of mutagenicity by catechin. AB - According to earlier studies, fumes from cooking oils were found to be mutagenic and several polycyclic aromatic hydrocarbons (PAHs), (benzo(a)pyrene (B(a)P), benz(a)antracene (B(a)A), and dibenz(a,h)anthracene (DB(ah)A)) were identified. Fume samples from three different commercial cooking oils frequently used in Taiwan were collected and nitro-polycyclic aromatic hydrocarbons (NPAHs) were extracted from the samples and identified by HPLC chromatography. Extracts from three cooking oil fumes contained 1-nitropyrene (1-NP) and 1,3-dinitropyrene (1,3 DNP). Concentrations of 1-NP and 1,3-DNP were 1.1 +/- 0.1 and 0.9 +/- 0.1 micrograms/m3 in fumes from lard oil, 2.9 +/- 0.3 and 3.4 +/- 0.2 micrograms/m3 in soybean oil, 1.5 +/- 0.1 and 0.4 +/- 0.1 micrograms/m3 in peanut oil, respectively. The preventive effect of three natural antioxidants (gamma tocopherol (TOC), lecithin (LEC), and catechin (CAT)) for the reduction of mutagenicity and amounts of PAHs and NPAHs of fumes from cooking oils were evaluated. Mutagenicity of cooking oil fumes occurred, and the concentration of B(a)P were significantly reduced (p < 0.05), by adding CAT into cooking oils before heating. B(a)A, DB(ah)A, and two NPAHs were not detected when the concentration of CAT was 500 ppm in all three cooking oil fumes. These results indicate that fumes of cooking oils contained PAHs and NPAHs that may be a risk factor for lung cancer among cooks and the carcinogens could be reduced by adding the natural antioxidant, catechin. PMID- 9726002 TI - Effect of endogenous carotenoids and defective RpoS sigma factor on spontaneous mutation under starvation conditions in Escherichia coli: evidence for the possible involvement of singlet oxygen. AB - Under starvation conditions, a variety of stationary phase genes are up-regulated under the control of the stationary phase sigma factor RpoS including at least two peroxidases and a protective DNA binding protein Dps. Previous work suggested that the reversion to prototrophy of certain amino acid auxotrophs of Escherichia coli that occurs when the bacteria are starved of a required amino acid results from the accumulation of oxidative damage to guanine residues in DNA. We report here that three strains lacking RpoS are indistinguishable from wild type in their ability to undergo this starvation-associated mutation, suggesting that basal levels of catalase activity are more than adequate in these strains, and that the induction of catalases and other proteins controlled by rpoS does not contribute to the protection of the DNA, at least in cells starved in early stationary phase. In comparison, the introduction of a plasmid specifying the production of singlet oxygen scavengers (carotenoids) in stationary phase cells led to a roughly twofold reduction in mutant yield. The results suggest that singlet oxygen may be an important endogenously produced mutagen in resting cells. PMID- 9726004 TI - Atypical background somatic mutant frequencies at the HPRT locus in children and adults with Down syndrome. AB - People with Down syndrome are 10-30 fold more likely to develop leukemia than the normal population. To date, little is known regarding the molecular mechanisms underlying this phenomenon. We have previously demonstrated that the spontaneous somatic mutant frequency (Mf) at a reporter gene, hypoxanthine-guanine phosphoribosyl transferase (HPRT), from a normal population showed a strict age dependency with an exponential increase in Mf from birth to late adolescents with a subsequent linear 2-5% increase per year in adults. In this study, we compared HPRT Mf in children and adults with Down syndrome using the HPRT T-cell cloning assay. We determined the Mf at the HPRT locus in 27 subjects with Down syndrome from ages 6 months to 53.4 years. Results demonstrated that background somatic Mf at the HPRT locus in children and adults with Down syndrome are not dependent on age as seen in a normal control population. Results also show that adults with Down syndrome have a significantly lower Mf than normal adults, and that children with Down syndrome have a significantly higher Mf than normal children, although the latter appears to be due to a decreased cloning efficiency (CE). These observations demonstrate that the frequency of spontaneous somatic mutations in children and adults with Down syndrome are atypical compared to normal controls, and suggest that the genetic mechanisms associated with background somatic mutational events in children and adults with Down syndrome may be different. PMID- 9726005 TI - Study of the frequencies of CYP1A1 gene polymorphisms and glutathione S transferase mu1 gene in primary breast cancers: an update with an additional 114 cases. AB - We studied the polymorphisms m1 (Msp1 restriction site) and m2 (codon Val substitution) of CYP1A1 gene and the copy number of glutathione S-transferase mu1 (GSTM1) gene on 487 DNA of breast cancer primary tumours from Caucasian group. Tumours of patients aged 55 years and under at diagnosis presented a great proportion of wild m1 (-/-) genotype; 83.6% vs. 69.5% (p < 0.0006), and a higher percentage of copy number of GSTM1 equal or under one copy; 65.2% vs. 53.4% (p < 0.011) for older patients m1 and m2 variants are closely linked (p < 0.0000). Tumour with a low copy number of GSTM1 is correlated with high histological grading (p < 0.01) and high Cathepsin D concentrations (p < 0.02). The combinations of different genotypes showed that association wild m1 (-/-) genotype and copy number of GSTM1 inferior or equal to one copy is correlated with an early onset of breast cancer primary tumour 44% vs. 6.4% for m1 (-/+) or (+/+) genotype and copy number of GSTM1 superior to one (p < 0.0000). The CYP1A1 gene wild form seems to be associated with early cancer development in Caucasian patients. PMID- 9726006 TI - Frequencies of hprt mutant lymphocytes in marijuana-smoking mothers and their newborns. AB - Reports of increases in the prevalence of marijuana smoking, especially among young people, have led to concerns about possible genotoxic effects from marijuana use due to exposure to the mutagenic and carcinogenic agents present in marijuana smoke. Prior studies of the adverse health consequences of marijuana smoking, using disease outcomes, have sometimes been confounded by the fact that most marijuana smokers also smoke tobacco. In the present study, the potential mutagenic effects of marijuana smoking were investigated with a somatic cell mutation assay that detects mutations occurring in vivo in the hprt gene. Subjects were volunteers recruited from a prenatal clinic that performs urine drug screens on all consenting patients. Blood samples were collected from 17 subjects whose drug screens indicated marijuana use, but who did not smoke tobacco or use cocaine or opiates, and 17 non-smokers with negative drug screens. Absence of tobacco use was confirmed by plasma cotinine tests. Cord blood samples were collected from newborns of 5 of the marijuana smokers and 5 non-smokers. Lymphocytes were isolated, cryopreserved, and later thawed and assayed with the autoradiographic hprt assay. The frequency of variant (mutant) lymphocytes (Vf) in the 17 non-smokers (+/- standard error) was 1.93 (+/- 0.17) per million evaluatable cells. The Vf of 17 marijuana smokers was more than three-fold higher, 6.48 (+/- 0.48) x 10(-6), a significant difference, p < 0.001. Cord blood lymphocytes from 5 newborns of non-smokers had a Vf of 0.85 (+/- 0.23) x 10(-6), compared to 2.55 (+/- 0.60) x 10(-6) for 5 newborns of marijuana smokers, significantly higher, p < 0.05. Because of the known association between increases in somatic mutations and the development of malignancies, this study indicates that marijuana smokers may have an elevated risk of cancer. For pregnant marijuana smokers, there is also concern for the possibility of genotoxic effects on the fetus, resulting in heightened risk of birth defects or childhood cancer. PMID- 9726007 TI - Free radicals generated in yeast by the Salmonella test-negative carcinogens benzene, urethane, thiourea and auramine O. AB - A large fraction of carcinogens score negative in short-term genotoxicity assays such as the Salmonella reverse mutation (Ames) assay. More information is needed about the mechanism of action of such Salmonella-negative carcinogens. Many Salmonella-negative carcinogens induce deletions due to intrachromosomal recombination in Saccharomyces cerevisiae with an apparent threshold. We have previously shown that the Salmonella-negative carcinogens cadmium, aniline, chloroform and carbon tetrachloride generate free radical species in S. cerevisiae. We have further investigated the possible generation of intracellular free radical species by the diverse Salmonella-negative carcinogens benzene, urethane, thiourea and auramine O. The toxicity and recombinagenicity of thiourea and auramine O was reduced in the presence of the free radical scavenger N-acetyl cysteine. N-acetyl cysteine did not protect against toxicity or recombination induced by the Salmonella-positive carcinogens ethyl methane sulfonate, methyl methane sulfonate or nitroquinoline-N-oxide. A strain deficient in the enzyme superoxide dismutase, which catalyses the dismutation of superoxide anion radical, was hypersensitive to killing by benzene, urethane and thiourea. The sod strain was only slightly more sensitive to the Salmonella-positive carcinogens. Intracellular oxidation of the free radical-sensitive reporter compound dichlorofluorescin diacetate was increased in yeast cultures exposed to benzene, urethane and auramine O; again, the Salmonella mutagens had no effect on oxidation of the dye. These data show that free radical species are produced in Saccharomyces cerevisiae following exposure to benzene, urethane, thiourea and auramine O, and suggest a possible role for oxidative stress is recombination induced by these carcinogens. PMID- 9726008 TI - AT-rich sequences flanking the 5'-end breakpoint of the 4977-bp deletion of human mitochondrial DNA are located between two bent-inducing DNA sequences that assume distorted structure in organello. AB - The 4977-bp deletion is the most common deletion among more than 90 large-scale deletions of human mitochondrial DNA (mtDNA) that are associated with aging and mitochondrial myopathies. The reason why the frequency of occurrence of this common deletion is so high in aged and myopathic human tissues is not clear. Since several studies proved that unusual DNA structures play very important roles in a number of recombination events, we hypothesized that some kind of unusual DNA structure may flank the breakpoints of the 4977-bp mtDNA deletion. We used two-dimensional (2-D) gel electrophoresis to assess the mobility abnormalities of the PCR-amplified DNA fragments encompassing the sequences of nucleotide position (np) 7901 to 9058 of human mtDNA. The results showed that the sequences of np 7901-8732 and np 8251-9058 exhibited retarded and increased mobilities, respectively, and that the sequence of np 8285-8676 showed normal mobility in the 2-D gel. This indicates that the 5'-end breakpoint of the 4977-bp deletion is located within the junction site of two flanking bent-inducing DNA sequences. We confirmed this notion by using osmium tetroxide (OsO4) to probe mtDNA in organello. The results showed that the two AT-rich sequences flanking the 5'-end breakpoint of the 4977-bp deletion are susceptible to OsO4 modification. These findings suggest that the DNA sequences of the 5'-end breakpoint of the common mtDNA deletion are rendered to assume a more distorted structure than B-DNA by these two flanking bent-inducing DNA sequences in organello and thereby render this region to be more vulnerable to attack by reactive oxygen species and free radicals. PMID- 9726009 TI - Radio-sensitive murine thymoma cell line 3SB: characterization of its apoptosis resistant variants induced by repeated X-irradiation. AB - 3SB, a mouse thymoma cell line, is one of the most radio-sensitive cells (D0 = 0.3 Gy), and its rapid apoptosis (4 h after 5 Gy irradiation, 90% apoptosis) seems to play a decisive role in enhancing the radiosensitivity. To understand the molecular mechanisms underlying extremely high radiosensitivity and rapid apoptosis, we attempted to isolate X-ray-resistant (XR) variants from 3SBH5, a stable subclone of 3SB, by repeating exposure of the cells to 2-5 Gy X-rays. Four independent stable XR variants, R111, R223, R316 and R429, were isolated by the repeated irradiation protocols. All XR cells possessed about 3 times higher D10 values than that of their parental 3SBH5. They were also resistant to apoptosis; only 10% cells underwent apoptosis 4 h after 5 Gy irradiation. The p53 protein was induced in all the cell lines after 5 Gy X-irradiation. These variants showed a cross resistance to a chemical reagent daunorubicin (DNR) that is known to be involved in the ceramide-mediated apoptosis. DNR, as well as C2-ceramide (5 muM) induced apoptosis in parental 3SBH5 cell, but not in two XR variants, R233 and R316 cells. Present result suggests that the induction of X-ray resistance by repeated X-irradiation might be achieved, at least partly, by the enhanced resistance to the ceramide-mediated apoptosis. PMID- 9726010 TI - Methods for registration of spontaneous DNA instability in mammalian cells. AB - A phenomenon of spontaneous DNA instability displays itself as the low level of repair DNA synthesis that takes place during any cell cycle phases. However, there is a problem in detection of very low intensive repair DNA synthesis. This paper suggests two approaches to detect the spontaneous DNA instability. The first method involves a blockade of the DNA gaps sealing by a combination of inhibitors, hydroxyurea and arabinofuranosyl cytosine. An accumulation of single strand gaps leads to production of DNA double strand breaks and results to reproductive inactivation of cells. It was shown that registration of both these events by different methods (such as viscoelastometry of DNA, orthogonal pulse electrophoresis or comet assay for double strand breaks as well as effectiveness of colony growth for cell inactivation) may be used as suitable measure of the spontaneous DNA instability. The second approach bases on photolysis of bromodeoxyuridine incorporated into repair DNA patches during the spontaneous repair DNA synthesis. Long wave UV irradiation of cells containing bromodeoxyuridine labeled DNA stained with Hoechst 33342 causes their inactivation. Experimental results presented confirm that both methods actually detect the spontaneous DNA instability. It takes note of the spontaneous DNA instability varies for cells from different tissues and species and increases during aging. PMID- 9726011 TI - Mutation analyses of KRAS exon 1 comparing three different techniques: temporal temperature gradient electrophoresis, constant denaturant capillary electrophoresis and allele specific polymerase chain reaction. AB - Mutations in the KRAS gene is a key event in the carcinogenesis of many human cancers and may serve as a diagnostic marker and a target for therapeutic intervention. In this study we have applied three different techniques for mutation detection of KRAS exon 1 mutations: Allele specific polymerase chain reaction (AS-PCR), temporal temperature gradient electrophoresis (TTGE) and constant denaturant capillary electrophoresis (CDCE). Samples from 191 sporadic colon carcinomas were analyzed. AS-PCR were performed with oligonucleotides specific for know mutations in codon 12 and 13 of the KRAS gene. In TTGE analyses, linear ramping of the temperature were performed during electrophoresis in a constant denaturant gel. CDCE analyses were performed using fluorescin labeled PCR-products. Separation was achieved under constant denaturing conditions using high temperature in a gel-filled capillary followed by laser detection. A mutated KRAS gene was found in 42/191 (22.0%) of the samples using AS-PCR, in 62/191 (32.5%) using TTGE and in 66/191 (34.6%) of the samples using CDCE. In the TTGE and CDCE analyses the sequence of the mutant were determined by comparing the electrophoretic pattern to that of known mutations or by mixing the sample with known mutations prior to reanalysis. In a titration experiment mixing mutant and wild-type alleles prior to PCR, the sensitivity for mutation detection was shown to be 10(-2) for TTGE and under optimized conditions 10(-3) for CDCE. PMID- 9726012 TI - A human lymphoid leukemia cell line with a V(D)J recombinase-mediated deletion of hprt. AB - Large deletions of exons 2 and 3 of the hprt gene are the most common type of hprt mutation in lymphocytes of newborn infants, and their frequency increases in cultured human T-lymphoid cells as a result of exposure to etoposide. Sequenced PCR products for these deletions are consistent with a V(D)J recombinase-mediated mechanism underlying their genesis. Herein, we describe the isolation and characterization of an etoposide-induced mutant CEM cell line that is clonal for a V(D)J recombinase-mediated exon 2 + 3 deletion. Human CCRF-CEM cells were exposed to 5 muM etoposide for 4 h, selected in 6-thioguanine, and an exon 2 + 3 deletion mutant was isolated through serial limiting dilution, using a PCR-based assay for detection of the exon 2 + 3 deletion. Untreated CEM cells and cells treated with 6-thioguanine alone were similarly subcultured. The exon 2 + 3 deletion-containing line was termed SJCEM808 and had a slightly longer doubling time than the control lines, tended to clump in suspension, and was characterized by cell membrane blebbing. Compared to the parent line, SJCEM808 had similar cytogenetic abnormalities, lower CD2, CD1, and CD10 expression, and negligible RAG-1 expression. However, RAG-1 expression was down-regulated in some untreated parental subclones following similar subculturing. The sequence of the exon 2 + 3 deletion mutation exhibited nucleotide insertions, and the breakpoints were adjacent to heptamer signal recognition sequences in intact hprt, consistent with a V(D)J recombinase-mediated mechanism underlying its genesis. There were no MLL gene or interlocus T-cell receptor (TCR) rearrangements. These results indicate that non-homologous recombination following etoposide treatment is neither necessarily accompanied by other large DNA rearrangements nor simply a pre-lethal event, and this cell line may serve as a useful tool for studying illegitimate V(D)J recombinase-mediated deletions. PMID- 9726013 TI - Oncoviral DNAs induce transposition of endogenous mobile elements in the genome of Drosophila melanogaster. AB - Previously, we have shown that particles of Rous sarcoma virus or cloned fragments of RSV cDNA as well as DNA of oncogenic simian adenovirus Sa7, injected into the polar plasm of early Drosophila melanogaster embryos, were able to induce, with high frequency, unstable visible mutations in different groups of genetic loci. The genetic instability of the recovered mutations, i.e., their ability to revert to normal state or to generate new mutant alleles at the affected locus, was manifest in mutant lines through several generations. The molecular analysis undertaken in this study of the yellow-scute loci region which is highly sensitive to the microinjected Sa7 DNA, and of the white locus, that frequently mutates under the influence of RSV cDNA, clearly shows that the induced mutations and reversions are accompanied by insertion/excision of endogenous mobile elements. This conclusion is confirmed by in situ hybridization experiments which demonstrate that the adenovirus DNA is able to change, though with different efficiency, the chromosomal localization of certain Drosophila retrotransposons. These results partially elucidate the molecular mechanism of the genetic instability in D. melanogaster induced by microinjection of oncoviruses into early embryos, implying that is results from mobilization of endogenous transposons which play the role of insertional elements directly causing unstable mutations. PMID- 9726015 TI - Induction by nitriles of sex chromosome aneuploidy: tests of mechanism. AB - Genetic and biochemical assays were conducted to determine if nitrile induced adult paralysis and germline aneuploidy in female Drosophila melanogaster requires a biochemical activation mechanism which results in the release of free cyanide. Two nitriles predicted to differ substantially in their susceptibility to enzymatic cyanide release were found to be equally effective inducers of aneuploidy. Regardless of differences in chemical structure, nitriles seem to be affecting a common cellular target as judged by the lack of synergistic effects when two nitriles are presented simultaneously. Mitochondrial respiration was not inhibited by acetonitrile under conditions in which sodium cyanide completely blocked respiration. A sensitive luciferase enzyme inhibition assay suggests that some, but not all, nitriles may affect hydrophobic protein interactions. These results suggest that there is no single biochemical mechanism by which all nitriles induce aneuploidy, although the cellular target disrupted is probably the same for each chemical. The implications of these findings for structural alert based pre-screening of mutagens are discussed. PMID- 9726014 TI - Mutation spectrum in the lacI gene, induced by gamma-radiation in aqueous solution under oxic conditions. AB - Irradiation of DNA in a cellular environment leads to many types of DNA damage, resulting from various effects of gamma-radiation. One of these effects is the formation of water-derived radicals (e.g., .OH radicals), which are formed in the vicinity of DNA (indirect effect). To study the influence of the indirect effect on gamma-radiation-induced mutations, a newly constructed plasmid, containing the lacI gene as a target gene, was irradiated with 60Co gamma-radiation in aqueous solution, in the presence of oxygen. Under these circumstances, only .OH radicals will be responsible for the induced mutations. Sequence analysis of the gamma radiation-induced mutations showed that 96% of all mutations were base pair substitutions, 87% of which occurred in the lacI gene, the others are formed in the lac operator part. All gamma-radiation-induced mutations in the lacI gene occurred exclusively on G:C base pairs, and no mutations at A:T base pairs could be detected. In the spontaneous mutation spectrum, 83% of all mutations were base pair substitutions, 35% of which occurred in the lacI gene and 48% in the lac operator part. Base pair substitutions on G:C base pairs were very similar in the gamma-radiation-induced and in the spontaneous mutation spectrum, implying a high contribution of .OH radicals to spontaneous mutagenesis. A:T to G:C transitions accounted for 10% of all spontaneous base pair substitutions in the lacI gene and are probably the result of effects, other than just .OH radicals. It can be concluded that .OH radicals are an important source for mutations at G:C base pairs. In this paper, the extracellular gamma-radiation-induced mutation spectrum is also compared to the previously obtained, intracellular gamma-radiation induced mutation spectrum of the lacI gene. Comparison shows some differences, such as relative high amounts of mutations at A:T base pairs, G:C to T:A transversions and frameshift mutations in the intracellular gamma-radiation induced mutation spectrum, as compared to the extracellular gamma-radiation induced mutation spectrum. Since the extracellular gamma-radiation-induced mutation spectrum shows that .OH radicals are mainly responsible for base pair substitutions on G:C base pairs, mutations at A:T base pairs in the intracellular gamma-radiation-induced mutation spectrum are apparently the result of additional or other factors. PMID- 9726016 TI - Sister chromatid exchange data and Gram-Charlier series. AB - Bowman et al. [K.O. Bowman, M.A. Kastenbaum, L.R. Shenton, Fitting multi parameter distributions to SCE data, Mutat. Res., 358 (1996) 15-24.] showed how discrete Pearson and discrete Johnson translation-system distributions may be fitted to sister chromatid exchange (SCE) data presented by Bender et al. [M.A. Bender, R.J. Pearston, R.C. Leonard, B.E. Pyatt, P.C. Gooch, On the distribution of spontaneous SCE in human peripheral blood lymphocytes, Mutat. Res., 281 (1992) 227-232.]. When their performances were measured by the chi-squared test of goodness of fit, these distributions proved to be only moderately better alternatives to the poorly fitting Poisson, binomial, and negative binomial distributions. In this paper, we extend our search for better characterizations of the SCE data by calling upon the Gram-Charlier type B approximation of the negative binomial distribution. We introduce an innovative extension of methods described in a little-known paper by Aitken and Gonin [A.C. Aitken, H.T. Gonin, On fourfold sampling with and without replacement, Proc. R. Soc. Edinburgh, 55 (1934) 114-125.], and show how this leads to fits of the SCE data that, in general, are within acceptable levels of probability. Moreover, we show how a theorem by Cramer [H. Cramer, Mathematical Methods of Statistics, Princeton Univ. Press, 1946.], relating to the scale factor m2/m'1 and its asymptotic distribution, may be used to discriminate between smokers and nonsmokers of the same gender. PMID- 9726017 TI - Development of long PCR techniques to analyze deletion mutations of the human hprt gene. AB - DNA primers and reaction conditions for long polymerase chain reaction amplification (PCR) of portions of the human hprt gene are presented. Use of these primers with DNA from previously defined hprt deletion mutants demonstrated production of the expected smaller size products as compared to DNA from wild type cells. These primers will be of value in the rapid analysis and subsequent sequencing of hprt deletion mutations in human cells. PMID- 9726018 TI - Increased radiation-induced chromosome breakage after progesterone addition at the G1/S-phase transition. AB - Pregnant females appear to have an increased chromosomal sensitivity to gamma irradiation. This hypersensitivity was found to parallel the increase of gestation hormone amounts [M. Ricoul, L. Sabatier, B. Dutrillaux, Increased chromosome radiosensitivity during pregnancy, Mutat. Res. 374(1997) 73-78]. An in vitro experiment was developed to study the effect of progesterone. We performed irradiations of whole blood from normal human donors and chromosome were analysed in first generation metaphases. By comparison to untreated controls, all cultures in which progesterone was added around the 24th h of culture exhibited an increased frequency of chromosome rearrangements, principally dicentrics and rings, which confirms the role of progesterone in the results of in vivo studies. BrdU incorporation studies suggested that progesterone was particularly efficient just before the entry into S-phase, which corresponds to the G1/S transition period. Cultures with an increased frequency of chromosome breakage had a slightly higher mitotic index than controls. It is suggested that progesterone may stimulate DNA repair in cells which reached the end of G1-phase with unrepaired breaks. This would allow the cells to enter the S-phase and survive, although some illegitimate repair leads to chromosome rearrangements, visible at the following metaphase. PMID- 9726019 TI - Chromosome abnormalities in peripheral blood lymphocytes from Macaca fascicularis and Erythrocebus patas (Cercopithecidae, Catarrhini) after X-ray irradiation. AB - In this paper, we describe the results of a qualitative and quantitative study of chromosomal reorganizations in X-irradiated (1 Gy and 2 Gy) lymphocytes from Macaca fascicularis (MFA) and Erythrocebus patas (EPA). A total of 515 breakpoints in M. fascicularis and 271 breakpoints in E. patas have been detected, identified and localized in the ideogram of the species. The Chi square test indicates that the distribution of breakpoints along the chromosomes is not random in M. fascicularis, and is not random for the p and q arms and bands in both species. Chromosome 5 of M. fascicularis (MFA5), chromosome 1 of E. patas (EPA1), and chromosome arms MFA5q, and EPA 1p are significantly more affected than expected, while chromosome MFA9 is less affected. Terminal regions of chromosome arms accumulate a higher number of breakpoints than the rest of the chromosome (44.4% in M. fascicularis and 45.98% in E. patas); 92.06% and 91.97% of breakpoints are observed in G negative bands in M. fascicularis and E. patas, respectively. PMID- 9726020 TI - Comparison of mutant frequencies and types of mutations induced by thiotepa in the endogenous Hprt gene and transgenic lacI gene of Big Blue rats. AB - The utility of the lacI transgene of Big Blue rats as a reporter of in vivo mutation was evaluated by comparing the frequency and types of mutations induced by thiotepa in the transgene and the endogenous Hprt gene. Transgenic rats were given i.p. injections of 1.4 mg/kg of thiotepa three times per week over a period of 4 weeks (a total dose of 16.8 mg/kg); 1 week after the last injection, mutation assays were performed on spleen lymphocytes isolated from the animals. Thiotepa treatment increased the lacI mutant frequency from 34.8 +/- 4.1 x 10(-6) in control animals to 140.9 +/- 64.8 x 10(-6) (p = 0.0020) and the Hprt mutant frequency from 3.5 +/- 1.5 x 10(-6) to 41.1 +/- 23.2 x 10(-6) (p = 0.0028). Sequence analysis of lacI mutant DNA and Hprt mutant cDNA produced similar overall mutation patterns: G:C-->T:A transversion was the most common base pair substitution (32% of independent mutations in the lacI gene and 28% of Hprt mutations), and deletions and insertions accounted for 34% of mutations in the lacI gene and 28% in the Hprt gene. The majority of thiotepa-induced base pair substitutions in the Hprt gene occurred with the mutated purine on the non transcribed DNA strand, while no strand-related bias was found for mutations in the lacI gene. Substitutions at G:C base pairs in the lacI gene, but not in the Hprt gene, were found disproportionately in CpG sites. In addition, multiplex polymerase chain reaction analysis of genomic DNA from the Hprt mutants indicated that 34% had relatively large deletions; none of these deletions was detected by the cDNA analysis. The results indicate that the frequency of thiotepa-induced mutants in Big Blue rats was 2.8-fold greater in the lacI gene than in the Hprt gene. Although the Hprt gene recovered a fraction of large deletions not found among the lacI mutants, the effects of transcription-coupled DNA repair in the Hprt gene and the targeting of base pair substitutions to G:C base pairs in CpG sites may have contributed to the higher mutant frequencies induced by thiotepa in the lacI transgene compared with the Hprt gene. PMID- 9726021 TI - Protease activation following UV irradiation is linked to hypomutability in human cells selected for resistance to combination of UV and antipain. AB - In order to examine the relationship between activation of an antipain-sensitive protease and suppression of mutability in UV (UVC)-irradiated human cells, a human cell variant with the high protease activity induced by UV was established and characterized for its susceptibility to UV-induced mutagenicity. Cells of a hypermutable cell strain, RSa, were mutagenized with ethyl methanesulfonate and irradiated with 10 J/m2 UV, followed by exposure to 20 mM antipain for 34 h. Whereas the combined treatment was totally lethal to RSa cells not treated with ethyl methanesulfonate, one surviving clone was isolated from the mutagenized cells and designated UVAP-1. When fibrinolytic protease activity was measured from extracts of the cell, it was found that the protease activity was elevated promptly after UV irradiation, reaching the maximum at 10 min post-irradiation. This protease activity was inhibited by antipain. After UV irradiation the phenotypic mutation frequencies of UVAP-1 cells were much lower than those of the parent RSa cells, as evaluated by the generation of clones resistant to ouabain killing. Furthermore, mutation at the K-ras codon 12 in genomic DNA was detected in RSa cells but not in UVAP-1 cells. Thus, the protease activation was correlated with the decreased levels of UV-mutagenicity in UVAP-1 cells, supporting the possible involvement of the antipain-sensitive protease activity in the regulation of cellular mutability following UV irradiation. PMID- 9726023 TI - Ras p21 protein levels in human plasma from patients with chronic obstructive pulmonary disease (COPD) compared with lung cancer patients and healthy controls. AB - To explore the value of an increase in ras p21 proteins in plasma as a biomarker for the carcinogenic process or for the general disease state, we have directly analysed for ras p21 proteins, plasma samples from Polish human patients with chronic obstructive pulmonary disease (COPD). They were compared with appropriate controls and also with the Polish lung cancer patients previously examined before treatment [D. Anderson, J.A. Hughes, A. Cebulska-Wasilewska, E. Nizankowska, B. Graca, Ras oncoproteins in human plasma from lung cancer patients and healthy controls, Mutat. Res. 349 (1996) 121-126]. An elevated level of ras p21 proteins was considered to be greater than 2 standard deviations (SD) above the mean negative control values. Nine out of 20 COPD patients (mean age = 65.9 years) had increased ras p21 protein levels when compared with 20 age-matched (mean age = 62.4 years) controls of the present study with a mean + 2 SD of 0.70. Eighteen out of 40 lung cancer patients (mean age = 60.1 years) had increased ras p21 protein levels compared with their concurrent controls (mean age = 40.2 years) with a mean + 2 SD of 2.53. However when compared with the age-matched controls of this present study, there were 35 out of 40 (87.5%) with increased levels. When the COPD patients and lung cancer patients were compared with 101 historical controls (age range 25-76 years, of those whose age was recorded) from unexposed healthy populations from Poland, Estonia and Spain with a mean + 2 SD of 1.83, then 4 out of 20 (20%) COPD patients and 30 out of 40 (75%) lung cancer patients had increased levels. Whether using concurrent controls, age-matched controls or historical controls, the data would suggest that an increase in ras p21 protein levels in plasma from lung cancer patients could be a possible prognostic marker or biomarker for lung cancer. COPD patients when compared with historical controls or age-matched controls had lower ras p21 protein values than cancer patients. Their ras p21 protein values might also be a biomarker for cancer. It is possible that some of these COPD patients were in the process of developing cancer or perhaps would die from COPD before cancer develops. It cannot be ruled out that the increases could be a biomarker of exposure since many of the lung cancer patients and most of the COPD patients were smokers. PMID- 9726022 TI - Genotoxicity of ribo- and deoxyribonucleosides of 8-hydroxyguanine, 5 hydroxycytosine, and 2-hydroxyadenine: induction of SCE in human lymphocytes and mutagenicity in Salmonella typhimurium TA 100. AB - The induction of SCE by ribo- and deoxyribonucleosides of 8-hydroxyguanine, 5 hydroxycytosine, and 2-hydroxyadenine was tested using human peripheral blood lymphocytes. All of these compounds caused an increase in SCE frequency. The potency of SCE induction was as follows: 5-OH-C, 5-OH-dC > 8.OH-G > 8-OH-dG > 2 OH-A, 2-OH-dA. These results suggest that the oxidized nucleosides are incorporated into DNA with different efficiencies (or are repaired with different efficiencies) and exhibit genotoxicity in human blood cells. Ribo- and deoxyribo derivatives of 5-OH-Cyt and 2-OH-Ade also showed mutagenic activity in Salmonella typhimurium TA 100. PMID- 9726024 TI - Correlation of UV-induced mutational spectra and the in vitro damage distribution at the human hprt gene. AB - We have determined the in vitro DNA damage distribution induced by 254 mm UV in the human hprt gene. The sequence-specific nature of the DNA damage for both main classes of UV-induced photoproducts, i.e., cyclobutane pyrimidine dimers (CPDs) and the pyrimidine <6-4> pyrimidone photoproducts (64PyPy), was evaluated. Utilizing an automated DNA sequencer plus auxiliary software, semi-automated analyses were performed for peak quantitation and retention-time to sequence position correlation. 64PyPy were predominantly formed at 5'-YTC-3' sites (p < 0.02; where Y = C,T). The effect of the 3'flanking nucleotide on the 64PyPy formation at 5'-TC-3' sites was 64PyPy at 5'-TCT-3' sites were induced at lower frequencies compared to 5'-TCM-3' sites (where M = A or C; p < 0.03). No effect of flanking nucleotides was detected for CPDs recovered at 5'-TT-3' sites. Sites of mutations in the hprt gene were compared to the sites of DNA damage. Two regions of frequently mutated nucleotides corresponded to sites of high deposition of damage. The two sites either had a high frequency of CPDs or 64PyPy, which implicated both types of photoproducts as premutagenic lesions. PMID- 9726025 TI - A theory for growth hormesis. PMID- 9726026 TI - Consideration of short-term carcinogenicity tests using transgenic mouse models. PMID- 9726027 TI - Why are there more mutations in males? PMID- 9726028 TI - The mutagenic potential of chlorothalonil: in vivo chromosome aberration studies. PMID- 9726029 TI - Hypotheses on the role of cytokines in peptic ulcer disease. AB - Helicobacter pylori is the cause of chronic type B gastritis and occurs in almost all patients with duodenal ulcers. Infection with H. pylori is characterized by an increased production of several inflammatory cytokines. Increasing evidence suggests a central role of these cytokines in the pathogenesis of H. pylori associated gastritis and peptic ulcer disease. Cytokines may be crucial in the recruitment and activation of inflammatory cells and in stimulation of gastrin release. In addition to their proinflammatory properties, cytokines may also inhibit the ulcer occurrence by stimulation of prostaglandins and somatostatin release and by direct impairment of acid secretion. The balance of these factors may determine the clinical outcome of H. pylori infection. PMID- 9726030 TI - Regulation of iron metabolism in the acute-phase response: interferon gamma and tumour necrosis factor alpha induce hypoferraemia, ferritin production and a decrease in circulating transferrin receptors in cancer patients. AB - BACKGROUND: The acute-phase response and anaemia of chronic disease are characterized by hypoferraemia associated with an increased ferritin synthesis, which might be mediated by the activated cytokine cascade. METHODS: We examined the prolonged effects of isolated limb perfusion (ILP) with recombinant human tumour necrosis factor alpha (rTNF), recombinant human interferon gamma (rIFN gamma) and melphalan on interleukin (IL) 6 and acute-phase protein levels, iron status and serum transferrin receptor (sTfR) levels in 12 patients with melanoma or sarcoma. Patients were treated with ILP during 90 min after pretreatment with rIFN-gamma during 2 days. RESULTS: After ILP, leakage of TNF resulted in systemic peak levels at 3 min followed by an increase in IL-6 with maximum levels at 4h. C reactive protein (CRP) rose at 4 h to peak levels at day 2, whereas alpha 1 antitrypsin and alpha 1-acid glycoprotein increased to maximum levels at day 3. Albumin and transferrin levels decreased after ILP and recovered after day 2. Serum iron and sTfR levels decreased during pretreatment and after ILP to minimum levels at 8 h and day 1 respectively. This was associated with an increase in serum ferritin levels, which paralleled CRP values. CONCLUSIONS: Our data point to a central role for the cytokine network in the modulation of iron metabolism in the acute-phase response and anaemia of chronic disease. TNF, possibly via induction of IL-6, and IFN-gamma induce hypoferraemia, which may in part result from a decrease in tissue iron release based on a primary stimulation of ferritin synthesis. The fall in sTfR levels may reflect an impaired erythroid growth and/or TfR expression mediated by TNF and IFN-gamma. PMID- 9726031 TI - Gelatinases in drug-induced toxic epidermal necrolysis. AB - BACKGROUND: The matrix metalloproteinases (MMPs) MMP2 and MMP9 play a significant role in epidermal detachment, inflammation and re-epithelialization. We have evaluated their activity in toxic epidermal necrolysis (TEN). DESIGN: The level and pattern of activity of MMP2 and MMP9 were investigated by measuring the degradation of 3H-labelled gelatin and by zymography in blister fluid from six TEN patients and compared the results with three other blistering conditions: bullous pemphigoid (n = 6), second-degree burn (n = 13) or suction blister (n = 3). RESULTS: A higher amount of MMP2 was found in TEN blister fluid with the constant presence of a significantly larger proportion of the activated forms of MMP2, a particular feature of TEN, than the other blistering diseases studied. CONCLUSION: This study emphasizes the potential role of MMP2 in the specific inflammatory reaction and reparation process in TEN skin. PMID- 9726032 TI - Lipoprotein (a) induces angiogenesis on the chick embryo chorioallantoic membrane. AB - BACKGROUND: Both lipoprotein (a) [Lp(a)] and angiogenesis have been shown to be associated with initiation and progression of atherosclerotic plaque. Lp(a) and two neutralizing anti-Lp(a) antibodies were investigated for their capacity to affect the vasoproliferative processes of the chick embryo chorioallantoic membrane (CAM), a useful model for such an investigation. METHODS: Gelatin sponges loaded with Lp(a) alone or together with anti-Lp(a) antibodies, or with vehicle alone, phosphate-buffered saline (PBS), were implanted in vivo onto the CAM at incubation day 8. Four days later, sponges and the adjacent CAM tissues were assessed for the extent of angiogenesis in terms of microvessel counts. RESULTS: Lp(a)-loaded sponges gave significantly higher counts than those loaded with the LP(a)-anti-Lp(a) antibodies complex, which overlapped those treated with PBS. The angiogenic response was similar to that obtained with basic fibroblast growth factor, a well known angiogenic molecule. CONCLUSION: These data suggest that Lp(a) is capable of inducing angiogenesis in vivo, which might account for its ability to enhance and support atherosclerosis. PMID- 9726033 TI - Tumour necrosis factor beta alleles and hyperinsulinaemia in coronary artery disease. AB - BACKGROUND: Hyperinsulinaemia and dyslipoproteinaemia are markers and risk factors for coronary artery disease (CAD) and non-insulin-dependent diabetes mellitus (NIDDM). We investigated the influence of a tumour necrosis factor beta (TNF-beta) gene polymorphism on serum parameters related to these metabolic disorders in patients with CAD. METHODS: A total of 199 patients with CAD and 81 control subjects with angiographically normal coronary arteries were studied. A digestion of amplified DNA with NcoI revealed three fragment patterns: homozygosity for TNF-beta *1 or TNF-beta *2 and heterozygosity (TNF-beta *1/*2). RESULTS: Patients with CAD who had increased serum insulin or C-peptide (fasting and after glucose load) were predominantly heterozygous for TNF-beta (72% vs. 47%) and less frequently homozygous for TNF-beta *2 (22% vs. 43%, P = 0 x 0.03). CONCLUSION: This study demonstrates an association of TNF-beta alleles with the risk factor hyperinsulinaemia in CAD. Genomic variants of TNF-beta may therefore contribute to the complex susceptibility for the metabolic syndrome in patients with CAD. PMID- 9726034 TI - Functional and structural properties of Na+/K(+)-ATPase enzyme in neonatal erythrocytes. AB - BACKGROUND: The Na+/K(+)-pump is the main regulator enzyme of intracellular monovalent cation concentration. There are only limited data available concerning its structure and function in healthy neonates, in comparison with data available regarding its structure and function in children. PATIENTS AND METHODS: Samples of 100 microL of anticoagulated blood were taken from 53 healthy neonates (age under 6th postnatal day, median age 3.5 days) and 61 healthy children (median age 12.4 months, range 6-36 months,). The Na+/K(+)-ATPase activity, its sensitivity to ouabain (a digoxin-analogue substance) and the expression of Na+/K(+) ATPase subunit isoforms were determined. RESULTS: The enzyme activity (429.2 +/- 17.2 versus 295.5 +/- 10.2 U, P < 0 x 0.001) and I50 value for ouabain inhibition (1.50 +/- 0.10 versus 0.96 +/- 0.10 mumol L-1, P < 0.05) was higher in neonates. More alpha 1 subunits (relative density: 1.16 +/- 0.10 versus 0.75 +/- 0.03, P < 0.001) and higher alpha 1/ alpha 2 ratio (4.14 +/- 0.21 versus 2.02 +/- 0.16, P < 0.01) were detected. CONCLUSION: This is the first study demonstrating changes of Na+/K(+) -ATPase molecules not only in enzyme activity, but also on protein level. Our results might contribute to the understanding of the resistance of neonatal cell membranes toward the pharmacodynamic actions of cardiac glycosides. PMID- 9726035 TI - Factors influencing the effects of murine cytomegalovirus on the pancreas. AB - BACKGROUND: As human cytomegalovirus (HCMV) infections are implicated in insulin dependent diabetes mellitus (IDDM), the effects of murine (M)CMV infection of inbred mice on the pancreas are of interest. RESULTS: Inflammation and periacinar oedema peaked on day 3 and were replaced by a focal inflammation, but infected cells were rare. The islets were spared in C57BL mice. Insulitis normally seen in non-obese diabetic (NOD) mice was accelerated, but infected NOD mice did not become glycosuric. Isotypes of total and autoreactive antibodies suggested a shift to a Th 1 response (IgG2a) in all MCMV-infected mice. MCMV-induced pancreatitis was not affected by MHC genes but was similar or less severe in BALB/c mice. As these lack the Cmv1 gene, which provides a protective natural killer (NK) cell response in C57BL congenic mice, the C57BL background may carry a pancreatitis susceptibility gene able to counter NK-mediated restriction of viral replication. Consistently, congenic mice expressing Cmvl on a BALB/c background did not display pancreatitis, unless depleted of NK cells. In vivo treatment with soluble cytokine receptors suggested that interleukin 1 (IL-1) and/or tumour necrosis factor alpha contribute to acinar necrosis in C57BL mice. PMID- 9726036 TI - Adenine nucleotides modulate epithelial wound healing in vitro. AB - BACKGROUND: Adenine nucleotides have been demonstrated to enhance structural and functional regeneration in experimental renal injury in rats. The mechanism of adenine nucleotide action have not been elucidated. The aim of this study was to characterize the effects of adenine nucleotides on intestinal epithelial wound healing in vitro. METHODS: The effects of adenine nucleotides on cell migration, cell proliferation and cell adhesion were studied in the non-transformed small intestinal epithelial cell line IEC-6 using an in vitro wounding model, a colorimetric BrdU assay and a hexosaminidase adhesion assay. RESULTS: The adenine nucleotides ADP and ATP were found to significantly stimulate epithelial cell restitution (migration) in vitro. Stimulation of epithelial restitution averaged 42% for ADP and 57% for ATP. In addition, adenine nucleotides inhibited the proliferation of rat small intestinal epithelial cells, averaging 56% for ADP and 74% for ATP. Enhancement of intestinal epithelial restitution and inhibition of epithelial cell proliferation by adenine nucleotides were mediated through transforming growth factor (TGF)-beta-independent pathways. CONCLUSION: These findings suggest that adenine nucleotides exert functional effects on intestinal epithelial cell populations and may play a role in the morphogenesis of the gastrointestinal tract and its remodeling after injury. PMID- 9726037 TI - Inhibition of platelet-vessel wall interactions by thromboxane receptor antagonism in a human in vitro system: potentiation of antiplatelet effects of aspirin. AB - BACKGROUND: Pharmacological inhibition of arachidonic acid metabolism has proven therapeutically useful in the prevention of cardiovascular events. METHODS: We have investigated the ability of Bay u 3405, a synthetic thromboxane antagonist, to interfere with platelet aggregation and arachidonic acid metabolism. The antiplatelet action was also analysed in a perfusion system in which vascular subendothelium was exposed to circulating human blood (10 min; shear rate = 800 s 1). Platelet interactions were morphometrically analysed and results compared with those obtained in studies with blood from donors taking aspirin (acetylsalicylic acid, ASA) (500 mg day-1). The additional effect of Bay u 3405 on the antiplatelet action of ASA was also evaluated. RESULTS: Bay u 3405 caused a dose-dependent inhibition of platelet aggregation induced by U46619 with a maximal effect at concentrations > or = 0.01 microgram mL-1. Higher concentrations (> or = 0.05 micrograms mL-1) also inhibited aggregations induced by ADP or collagen. Bay u 3405 did not interfere with platelet arachidonic acid metabolism. In perfusion studies, Bay u 3405 (0.01 microgram mL-1) significantly decreased the total surface of the vessel covered by platelets (%CS = 18.7 +/- 1.09 vs. 24.4 +/- 1.94; P < 0.05) and the formation of large aggregates %T = 7.5 +/- 0.87 vs. 19.3 +/- 1.61; P < 0.01). ASA treatment reduced platelet aggregate formation (%T = 13.7 +/- 2.06; P < 0.05) but did not affect the total surface covered by platelets. The in vitro addition of Bay u 3405 to blood from ASA treated donors further reduced the formation of large aggregates (%T = 2.7 +/- 0.79; P < 0.01 vs. ASA). CONCLUSIONS: In vitro effect of Bay u 3405 on platelet function were superior to those observed with ASA. The thromboxane antagonism antagonism provided by Bay u 3405 further enhanced the inhibition of platelet aggregate formation found after ASA treatment. PMID- 9726038 TI - In vitro culture growth of erythroid progenitors and serum erythropoietin assay in the differential diagnosis of polycythaemia. AB - BACKGROUND: We assessed the in vitro culture growth of erythroid progenitors [burst forming unit-erythroid (BFU-E)] and serum erythropoietin (EPO) levels in different groups of polycythaemia to determine the discriminative power in differential diagnosis of polycythaemia. METHODS: We used the methylcellulose culture technique to study the growth of endogenous erythroid colonies (EECs) and EPO-dependent BFU-E from bone marrow (BM) and/or peripheral blood (PB) cells from 40 patients with polycythaemia vera (PV), 13 with secondary polycythaemia (SP), 19 with pure erythrocytosis (PE), five with PE and PV evolution later (PE-PV), and 12 with relative polycythaemia (RP). The serum EPO levels were measured by radioimmunoassay before treatment in 47 patients, 23 SP patients, 19 PE patients, five PE-PV patients and 16 RP patients, as well as after treatment in 38 PV patients, five PE-PV patients and 12 PE patients. RESULTS: The results of the erythroid progenitor culture assay showed that the numbers of EPO-dependent BFU-E in BM did not differ significantly among groups. The PB BFU-E were significantly higher in PV than in SP or PE, and no statistical difference were found among patients with SP, PE and RP. There was a correlation between BM BFU-E and PB BFU E in the individual PV and PE patients. EECs were present in all BM and PB cultures of untreated and phlebotomy-treated PV and PE-PV patients, but were absent in 6 of 17 PV patients who had received cytotoxic therapy. EECs were not found in SP, PE and RP. PB could substitute for BM in the EEC or the BFU-E assay. Both pretreatment and post-treatment serum EPO levels of PV and PE-PV were similar, which were significantly lower than SP, PE or RP. The serum EPO levels in treated PV or PE-PV patients who had normal haematocrit values were not significantly different from those in untreated patients. In contrast, the phlebotomy-treated PE patients had significantly higher serum EPO values than untreated PE patients. In the differentiation between PV and PE, the sensitivity, specificity, positive predictive value and negative predictive value of post treatment serum EPO levels at a cut-off level of < or = 9 UL-1 were 74%, 92% and 52% respectively. The discriminative power of post-phlebotomy serum EPO levels was even higher with a positive predictive value of 80% and negative predictive value of 92% for the prediction of PV evolution in patients with pure erythrocytosis of unknown origin. CONCLUSION: The present study showed that apart from EEC assay, the post-phlebotomy serum EPO level was a sensitive and specific parameter in the differential diagnosis of polycythaemia, in particular for the identification of PV among patients with unclassifiable polycythaemia. PMID- 9726039 TI - Effect of S-adenosylmethionine versus tauroursodeoxycholic acid on bile acid induced apoptosis and cytolysis in rat hepatocytes. AB - BACKGROUND: S-adenosylmethionine (SAMe) increases survival in alcoholic liver cirrhosis and may have a beneficial effect in cholestatic liver disease. SAMe repletes glutathione stores and protects tissue from oxygen free radicals. The effect of SAMe on bile acid-induced apoptosis is unknown. In the present study the possible hepatoprotective effect of SAMe was evaluated and compared with that of tauroursodeoxycholic acid (TUDCA). METHODS: Primary rat hepatocytes treated with glycochenodeoxycholic acid (GCDCA) were used as a model for cholestasis induced hepatocellular damage, which served to study the effects of SAMe and TUDCA on bile acid-induced apoptosis and cytolysis. RESULTS: SAMe reduced bile acid-induced apoptosis but did not prevent bile acid-induced cytolysis. Compared with SAMe, TUDCA was more efficient in reducing apoptosis due to toxic bile acids. The combination of SAMe and TUDCA had additive effects in reducing apoptosis. CONCLUSION: The reduction in bile acid-induced apoptosis by SAMe may represent one of the factors responsible for its beneficial effects in the treatment of liver diseases. PMID- 9726040 TI - The effect of smoking on post-heparin lipoprotein and hepatic lipase, cholesteryl ester transfer protein and lecithin:cholesterol acyl transferase activities in human plasma. AB - INTRODUCTION: Smoking is associated with dyslipidaemia, particularly raised plasma triglycerides and reduced high-density lipoprotein (HDL)-cholesterol and a delayed clearance of triglyceride in fat tolerance tests. The aim of this study was to investigate whether these phenomena could be explained by a reduced lipoprotein lipase activity in smokers. METHODS: A group of 40 healthy individuals [plasma cholesterol 5.07 (SD 0.90) mmol L-1, plasma triglyceride 1.02 (SD 0.39) mmol L-1)] was studied to examine the effects of smoking on plasma enzyme activities, particularly post-heparin lipase activities. The group comprised 20 smokers and 20 non-smokers, who were matched for age, gender and body mass index (BMI). RESULTS: Post-heparin lipoprotein lipase (LPL) activity [3.89 (SD 1.58) vs. 5.85 (SD 2.30) mumol free fatty acids (FFA) mL-1 h-1, P < 0.005], but not post-heparin hepatic lipase (HL) activity, was reduced in smokers. Plasma cholesteryl ester transfer protein (CETP) activity and lecithin: cholesterol acyl transferase (LCAT) activity were measured in a subgroup of 18 individuals, comprising nine smokers with nine matched non-smokers. There was no difference in CETP activities between two groups, but smokers had a significantly reduced plasma LCAT activity [112 (SD 23) vs. 152 (SD 24) nmol cholesterol mL-1 h 1, P < 0.005]. In both smokers (r=-0.53, P < 0.05) and non-smokers (r=-0.54, P < 0.05), HDL2 concentration was negatively associated with HL activity. In non smokers, HDL3 concentration was negatively associated with CETP activity (r= 0.77, P < 0.05), whereas in smokers HDL3 concentration was negatively associated with LCAT activity (r= -0.78, P < 0.050). CONCLUSION: It was shown by direct measurement that the activity of plasma post-heparin LPL is reduced in smokers, independently of age, gender and BMI. It is concluded that this enzyme perturbation associated with smoking may contribute to the development of the atherogenic lipoprotein phenotype seen in smokers. PMID- 9726042 TI - Endolymphatic sac tumours. AB - This review article surveys clinical and pathological literature on endolymphatic sac tumours (ELST) and summarizes characteristics that describe the entity. ELST are rare neuroectodermal neoplasms in the petrous bone, originating from inner ear structures. They can be encountered sporadically or in von Hippel-Lindau disease. The most prominent symptom is sensorineural deafness. Historically, nomenclature of invasive adenoid tumours in the petrous bone has been divergent, the term papillary adenocarcinoma used most frequently. Histologically, they have a follicular or papillary and adenoid pattern that can be easily confused with various other neoplastic conditions including metastatic carcinoma. It remains to be verified whether similar tumours (papillary adenocarcinomas) can originate from the middle ear. Middle ear adenomas have a similar appearance but probably originate from neural crest cells in the middle ear. ELST can express a variety of epitopes (including cytokeratin and neuroectodermal markers) which can be detected immunohistochemically. In cases in von Hippel-Lindau disease the cerebello-pontine angle should be included in routine radiological examinations to detect ELST before the tumours lead to deafness. In apparently sporadic cases of ELST, genetic testing for von Hippel-Lindau disease should be considered. Correct distinction of ELST from metastatic carcinoma prevents futile searches for unknown primary tumours. PMID- 9726041 TI - Gastrin/cholecystokinin type B receptors in the kidney: molecular, pharmacological, functional characterization, and localization. AB - BACKGROUND: Gastrin/cholecystokinin type B receptors (CCKBRs) can be found on parietal cells and smooth muscle cells and are the predominant brain CCK receptors. Recent cloning studies indicate that this is receptor type might also be expressed in the kidney. MATERIALS AND METHODS: We used Northern blot analysis in guinea pig. kidney and reverse transcriptase polymerase chain reaction (RT PCR) in several murine kidney cells lines to evaluate this organ for the expression of the CCKBRs. The receptor was pharmacologically characterized by displacement experiments using [125I]-BH-CCKs and various agonists and antagonists. Polyclonal antibodies vs. the CCKBRs were raised in chicken, and immunohistochemistry on tissue sections was used to localize the receptor within the organ. The effect of gastrin on renal cell growth was measured using proximal tubulus (MCT) cells, which were cultured with gastrin (10-9 M) for 24-72 h. Cell counts and [3H]-thymidine incorporation experiments were performed. RESULTS: CCKBR transcripts can be detected in kidney RNA (tubules > glomeruli > interstitium). RT-PCR revealed CCKBR transcripts in proximal tubules (MCT cells) and in mesangium (MMC). The medullary thick ascending limb of Henle's loop and several control tissues such as liver and muscle were negative. Displacement experiments using [125I]-BH-CCK and various agonists and antagonists identified binding sites with typical CCKBR pharmacology. CCKBRs were localized in the proximal tubulus, distal collecting ducts and mesangium cells. Treatment of rested MCT cells with gastrin 17-1 induced cell proliferation and [3H]-thymidine incorporation by at least 40% compared with normal growth (P < 0.05). CONCLUSION: These results show for the first time that CCKBRs are present in selected areas of the kidney, and strongly confirm our previous observation that this organ expresses binding sites for [125I]-gastrin. Furthermore, gastrin might act as a growth factor in the kidney. PMID- 9726043 TI - De-differentiated chondrosarcoma is not a 'de-differentiated' chondrosarcoma. AB - AIMS: De-differentiated chondrosarcoma is characterized by the presence of two distinct chondroid and nonchondroid tumour portions. The aim of our study was to investigate the distribution of extracellular matrix components in this tumour entity and thus to shed light on its histogenetic origin. METHODS AND RESULTS: Histochemical and immunohistochemical analyses were performed for collagen subtypes I, II, III and VI and cartilage proteoglycans in three samples of de differentiated as well as conventional chondrosarcomas (various grades). In the chondroid tumour areas of de-differentiated chondrosarcoma, typical cartilage matrix components could be detected similar to chondroid areas of grade 1 and 2 conventional chondrosarcomas. In contrast, the tumour matrix of the nonchondroid portions of de-differentiated chondrosarcomas contained matrix molecules which are typical for fibroblastic tissue. This matrix composition was not identical with less differentiated (nonchondroid) areas of grades 2 and 3 conventional chondrosarcomas. CONCLUSIONS: Our results confirm the chondroid nature of the differentiated portion of de-differentiated chondrosarcoma and indicate a nonchondrocytic nature of the nonchondroid portion. De-differentiated chondrosarcoma should not be considered as a 'de'-differentiated chondrosarcoma (grade 4 neoplasm), but as a tumour entity showing two types of mesenchymal differentiation. PMID- 9726044 TI - Angiomyolipoma predominantly composed of smooth muscle cells: problems in histological diagnosis. AB - AIMS: Five cases of angiomyolipoma (AML) composed exclusively or predominantly of smooth muscle cells (SMC) are presented to emphasize the histological diversity and to caution against errors in histological diagnosis. METHODS AND RESULTS: Four tumours were located in the liver and one in the renal capsule. Three patients were female and two were male, ranging from 36 to 76 years of age with a mean age of 50 years. One patient with a renal capsular tumour was associated with tuberous sclerosis. Two tumours were composed predominantly of a spindle shaped SMC component, whereas three others were composed predominantly of epithelioid SMC elements. AMLs composed exclusively or predominantly of spindle shaped smooth muscle cells (SMCs) stimulated leiomyoma, whereas AMLs composed exclusively or predominantly of epithelioid SMCs resembled epithelioid leiomyoma or leiomyosarcoma or other sarcoma when cellular atypia was present. However, both spindle and epithelioid SMCs were characteristically positive for HMB-45 melanoma-specific antibody; no other tissue components in either the liver or kidney were reactive to HMB-45. CONCLUSIONS: AML is often composed predominantly of SMC elements, and morphological features of the SMC elements are quite variable. Therefore, careful attention must be given to histological assessment of AML. Whenever a pathologist encounters an unfamiliar hepatic or renal tumour, the possibility of AML should be considered. Reactivity for HMB-45, however, confirmed the diagnosis of AML. PMID- 9726046 TI - Altered contractile proteins and neural innervation in idiopathic megarectum and megacolon. AB - AIM: The aetiology of idiopathic megarectum and idiopathic megacolon is unknown. We postulated that biochemical or ultrastructural abnormalities may be pathologically important, as has been observed in patients with chronic idiopathic intestinal pseudo-obstruction. METHODS AND RESULTS: Resection specimens from five patients with idiopathic megarectum or megacolon were processed for paraffin wax-embedded tissue histology (haematoxylin and eosin, Gomori trichrome and picrosirius stains, phosphatase activity and periodic acid Schiff staining), and frozen tissue for histochemistry and electron microscopy. The antibodies used in the immunohistochemistry were to myosin light chain kinase, smooth muscle myosin, alpha and beta actins, filamin, tropomyosin, phosphorylated and dephosphorylated neurofilaments and N-CAM. Variable hypertrophy of the muscularis mucosae and externa, no atrophy, and a variable nerve density decrease in longitudinal muscle and increase in the lamina propria was seen. In one patient beta actin and myosin light chain kinase immunoreactivity was reduced. CONCLUSION: Variable changes in innervation, and an abnormal contractile protein immunoreactivity pattern in one patient, may be of pathogenic importance. These clinically homogeneous conditions are likely to be due to a range of underlying pathogenic abnormalities. A search for further specific biochemical abnormalities is justified. PMID- 9726045 TI - Value of histochemical stains for copper in the diagnosis of Wilson's disease. AB - AIMS: The histochemical demonstration of hepatic copper is important in the diagnosis of Wilson's disease (WD). Conflicting results have been published with regard to the ability of different histochemical methods to demonstrate copper storage in the liver. Therefore, we evaluated the diagnostic value of three available histochemical methods in a large series of patients affected by WD. METHODS AND RESULTS: Seventy-nine consecutive liver needle biopsies, from 74 patients, 39 males and 35 females, aged 4-60 years (mean age 28.5 years) were stained with orcein, rhodanine and using Timm's method. On the basis of the histological picture, liver biopsies were subdivided into three groups: group A, steatosis; group B, interface hepatitis; group C, chronic hepatitis with bridging fibrosis and/or cirrhosis. In group A, 30.4% of the cases were positive using Timm's method, vs 13.2% using the rhodanine and 17.5% using the orcein method. In group B, Timm's method was positive in 40.1% while rhodanine and orcein showed positivity in 26.7%. In group C, the Timm's method stained 58.6%, rhodanine 36.6% and orcein 29.3% positively. CONCLUSIONS: Our data show that: (1) Timm's silver stain is the most sensitive method for the demonstration of copper in all cases of WD; (2) rhodanine and orcein have minor value in the diagnosis of WD, especially in the early stages of the disease; (3) to increase the diagnostic value of histochemistry for copper multiple histochemical stains in serial sections are required; and (4) although hepatic copper concentration is highest in the early stages of WD, the histochemical demonstration fails in a large number of cases. PMID- 9726047 TI - CD44v6 expression is related to progression in renal epithelial tumours. AB - AIMS: Expression of CD44 variant isoform including exon 6 has been associated to tumour progression in several carcinomas. However, no studies have been performed to assess the prognostic value of the expression of this marker in renal cell tumours. METHODS AND RESULTS: We studied 58 renal cell tumours. All patients were followed up for at least 3 years after nephrectomy. Tumours were analysed for expression of CD44v6 assessed by two isoform-specific monoclonal antibodies. RT PCR was performed to detect CD44 variant transcripts in 10 cases in which immunohistochemistry was negative. Twenty-two tumours showed reactivity in at least 1% cells for both antibodies with a strong membrane pattern. RT-PCR did not show CD44v6 transcripts in any of 10 studied tumours. Immunohistochemical staining was more frequent in perivascular areas or in areas of vascular invasion. In fact, CD44v6 expression correlated well with nuclear grade (P = 0.009), stage at diagnosis (P = 0.04) and appearance of metastasis after nephrectomy (P = 0.007). Although univariate survival analysis showed stage (P < 0.001), grade (P = 0.009) and CD44v6 expression (P = 0.04) to be significant predictive factors, only stage remained significant (P = 0.0013) in the multivariate analysis. CONCLUSIONS: CD44v6 expression, assessed immunohistochemically, is related to tumour progression. However, its prognostic value in renal cell tumours is dependent on tumour stage at diagnosis. PMID- 9726048 TI - Parathyroid glands in uraemic patients with refractory hyperparathyroidism: histopathology and p53 protein expression analysis. AB - AIMS: Refractory hyperparathyroidism is a state of parathyroid hyperfunction and hypercalcaemia in uraemic patients with previous secondary hyperplasia. We studied histopathological features and p53 expression in 49 parathyroid glands of uraemic patients (n = 21) with refractory hyperparathyroidism in order to investigate whether p53 abnormalities could be present in parathyroid hyperplasias of chronic renal failure. METHODS AND RESULTS: Nodular hyperplasia was found in 77.5% of the glands (n = 38). The proportion of oxyphil cells and acinar cell arrangements was higher in nodular hyperplasia than in diffuse hyperplastic glands P < 0.001). Duration of renal disease and haemodialysis treatment tended to be longer in patients with nodular hyperplasia. There was no correlation between serum intact PTH (iPTH), calcium and hyperplasia pattern. A trend for a higher glandular mass was found in nodular type hyperplasia (1.88 +/- 2.13 g) than in diffuse type hyperplasia (0.87 +/- 1.28 g; P = 0.06). Nuclear p53 immunoreactivity was shown in 55% of the hyperplastic glands, whereas it was not detected in 12 normal parathyroid glands used as controls. p53 staining was present in c. 82% of the diffuse hyperplastic glands and in 47% of the nodular hyperplastic glands (P = 0.08). CONCLUSIONS: Nodular type hyperplasia was the predominant histopathological pattern in uraemic patients with refractory hyperparathyroidism in our study. Nodular hyperplastic glands characteristically had higher percentage of oxyphil cells, acinar cell arrangements and mass than diffuse hyperplastic glands. A high prevalence of p53 protein expression was found in hyperplastic glands of uraemic patients. Our results suggest that p53 abnormalities might be involved in the pathogenesis of parathyroid hyperplasia in chronic renal failure. PMID- 9726049 TI - Multiple synchronous lung cancers and atypical adenomatous hyperplasia in Li Fraumeni syndrome. AB - AIM: We report a patient with Li-Fraumeni syndrome who developed multiple synchronous primary lung cancers together with atypical adenomatous hyperplasia, and discuss the pathogenesis of the lesions. CASE DETAILS: A 57-year-old woman with Li-Fraumeni syndrome and a history of smoking was diagnosed as having multiple synchronous primary lung tumours, all of which were treated by surgical excision. Histological examination showed three separate well-differentiated adenocarcinomas and a single focus of atypical adenomatous hyperplasia. A constitutional mutation of the p53 gene had been identified, both in this patient and other members of her family. CONCLUSION: Multiple synchronous primary lung tumours have not been reported before in association with this syndrome. With both smoking and Li-Fraumeni syndrome known to be associated with mutations in the p53 gene, a cumulative effect is suggested in this patient. PMID- 9726050 TI - Partial regression in primary carcinoma of the lung: does it occur? AB - AIMS: To study immunohistochemically a group of 28 primary lung cancers which demonstrated histological features reminiscent of those characteristic of regression in malignant melanoma, to determine the phenotype of their immune cell infiltrates. METHODS AND RESULTS: Using a standard s-ABC immunoperoxidase method then quantification by the Leica Q500 MC image analyser, the cellular infiltrate in these tumours, when compared to 67 control cases, showed excess CD3 + T lymphocytes (P < 0.007), with an increase in CD4:CD8 ratio and increased CD68 + macrophages (P = 0.00001). CD57 + natural killer cells and S100 + Langerhans cells were also increased, but not quite significantly (P = 0.048 and P = 0.072, respectively). CONCLUSIONS: This immunophenotype resembles that shown in regressing skin malignancies and suggests a similar process occurring in lung cancer. Regressing lung cancers are associated with a better prognosis than matched controls (P = 0.0034), some showed radiological evidence of growth retardation and the group had an excess of the large cell undifferentiated histological type. PMID- 9726051 TI - Apoptosis is associated with atypical or malignant change in meningiomas. An in situ labelling and immunohistochemical study. AB - AIMS: Although necrosis is an important phenomenon with implications for grading and prognostication in meningiomas, the alternate form of cell death, apoptosis, has not been extensively studied. In this series, we aimed to determine whether apoptosis in meningiomas correlated with histological types and grading. We also looked for a relationship between expression of apoptosis-related genes bcl-2, p53, c-myc and apoptosis meningiomas. METHODS AND RESULTS: Fifty-one meningiomas of diverse histological subtypes and grades were investigated with in situ end labelling of DNA fragments as well as immunohistochemical analysis of three apoptosis-related genes: p53, bcl-2 and c-myc. Our results showed that the apoptosis index was significantly higher in high-grade meningiomas (0.12%, n = 12) in than the benign meningiomas (0.023%, n = 39) P = 0.001) but there was no difference among the different histological subtypes of the benign meningiomas (P = 0.125). There is no obvious relationship between p53, bcl-2 and c-myc staining and apoptosis index in this group of meningiomas. CONCLUSION: We conclude that apoptosis is an important phenomenon in meningiomas and that it is associated with atypical or malignant changes in meningiomas. Apoptosis in meningiomas has no clearcut relationship with expression of p53, bcl-2 and c-myc. PMID- 9726052 TI - Spindle epithelial tumour with thymus-like element (SETTLE): the predominantly monophasic variant. AB - AIMS: To describe two cases of spindle epithelial tumour with thymus-like element (SETTLE) which are composed predominantly of spindle cells. In addition, to highlight some unusual histological features in SETTLE and discuss its separation from histological mimics. METHODS AND RESULTS: The thyroid masses were in a 4 year-old boy and a 25-year-old male. Both patients were euthyroid and were well except for thyromegaly. The specimens were formalin fixed, and immunohistochemistry was performed on this material using a panel of antibodies following microwave antigen retrieval. Morphologically, the dominant pattern was of sheets of spindle cells arranged in several patterns. There was mild pleomorphism and occasional mitoses. There were very small foci of squamous epithelium and occasional ductular structures. The stroma was composed of fibrous tissue and isolated areas of calcification were noted. The spindle cells showed strong and diffuse immunoreactivity with AE1/3, CAM5.2 and vimentin. CONCLUSION: We describe two cases of SETTLE that are composed mainly of spindle cells and only a very focal ductular component. In addition, calcification was noted within the stroma in one of the cases. These predominantly spindle examples of SETTLE must be separated from synovial sarcoma, which is a more mitotically active, aggressive tumour displaying only patchy immunopositivity with epithelial markers. PMID- 9726053 TI - Focal endometrial stromal hyperplasia. AB - AIMS: To describe a patient in whom multiple endometrial biopsies revealed stromal proliferative lesions causing diagnostic difficulty. METHODS AND RESULTS: A 39-year-old woman with a history of menorrhagia and dysmenorrhoea underwent two endometrial biopsies and hysterectomy. In each case histopathology revealed apparently multifocal benign stromal proliferative lesions with no evidence of invasive growth pattern. The lesions were characterized by increased cellularity, nuclear enlargement and spindle cell morphology but no mitotic activity or nuclear pleomorphism were seen. The possibility of low grade stromal neoplasm was considered in the biopsy specimens. The hysterectomy revealed no evidence of malignancy but foci of ovarian endometriosis and cervical stromal endometriosis were present. CONCLUSION: We suggest that the lesions represent focal endometrial stromal hyperplasia, a potential mimic of stromal nodule or stromal sarcoma in biopsy samples. PMID- 9726054 TI - Massive lymphocytic infiltration of uterine leiomyomas associated with GnRH agonist treatment. AB - AIMS: To report two unusual cases of massive lymphocytic infiltration of uterine leiomyomas, following GnRH agonist treatment. Previous reports have described a variety of alterations in leiomyoma histology following such therapy. METHODS AND RESULTS: These cases were studied using haematoxylin and eosin stains, and immunohistochemistry for B and T-cell markers (CD20/CD79a and CD3/UCHL-1) was performed. Leiomyomas were heavily infiltrated predominantly by mature lymphocytes of T-cell phenotype. Associated myocyte degenerative changes were present. CONCLUSIONS: Massive lymphocytic infiltration of leiomyomas may occur as a result of GnRH agonist treatment, although the mechanism is unclear. PMID- 9726055 TI - HPV typing and testing in gynaecological pathology: has the time come? PMID- 9726056 TI - Biotin inclusions: a potential pitfall in immunohistochemistry. PMID- 9726057 TI - bcl-2, p53 immunophenotypes and apoptosis in squamous cell carcinoma of the oesophagus. PMID- 9726058 TI - Osteoid and bone formation in a nasal mucosal melanoma and its metastasis. PMID- 9726059 TI - Atypical fibroxanthoma with granular cells. PMID- 9726061 TI - Bronchial epithelial-myoepithelial carcinoma. PMID- 9726060 TI - Synchronous occurrence of cutaneous lymphadenoma and syringoid eccrine carcinoma in a single patient. PMID- 9726062 TI - Epithelioid lymphangioleiomyomatosis-like tumour of the uterus in a patient without tuberous sclerosis: a lesion mimicking epithelioid leiomyosarcoma. PMID- 9726063 TI - Desmoplastic malignant melanoma of the uterine cervix: a rare primary malignancy in the uterus mimicking a sarcoma. PMID- 9726064 TI - Natural variation of cervical range of motion: a one-way repeated-measures design. AB - OBJECTIVE: To describe the natural variation of the active and passive cervical range of motion (ROM) in asymptomatic subjects over a 3-wk period. STUDY DESIGN: One-way repeated measures of active and passive cervical ROM. SETTING: Institute of Medical Biology (Center of Biomechanics) at Odense University, Denmark. PARTICIPANTS: Forty asymptomatic students from the University of Odense. Male/female ratio, 20:20; mean age, 23.9 yr (range, 20-30 yr). INTERVENTION: Measurements of the active and passive cervical ROM were taken using the electrogoniometer CA-6000 Spine Motion Analyzer. Each subject was measured six times during a 3-wk period. The measurements were performed at the same time of the day. The device gives the maximum end ROM for the motion plane examined. RESULTS: The natural variation in active and passive ROM was found to be in the order of chi +/- 20 degrees for flexion/extension, chi +/- 12 degrees for lateral flexion and chi +/- 14 degrees for rotation. CONCLUSION: In asymptomatic subjects, the individual natural variation is quite large for active and passive cervical flexion/extension, lateral flexion and rotation. When measuring individual patients, one should allow for a natural variation of 12-20 degrees. PMID- 9726065 TI - The relationship between posture and curvature of the cervical spine. AB - OBJECTIVE: To study the relationship between posture and curvature of the cervical spine in healthy subjects. SUBJECTS: The study was composed of 54 healthy students (25 men and 29 women) aged 20-31 yr with a mean age of 24.7 yr. METHODS: Lateral radiographs were taken of the head and cervical spine of the subjects while standing in a neutral position. Cervical spine posture was quantified by the angle of a reference line, composed of reference points of the upper six cervical vertebrae, with the horizontal axis. The curvature of the cervical spine was classified visually as lordotic, straight or reversed. RESULTS: A relationship was found between posture and curvature of the cervical spine (p = .006); a more forward posture of the cervical spine was related to a partly reversed curvature; and a more upright posture was related to a lordotic curvature. Moreover, men more often exhibited a straight curvature, and women more often exhibited a partly reversed curvature. CONCLUSION: The curvature of the cervical spine is related to the subject's posture and gender. PMID- 9726067 TI - Spinal manipulation in China. AB - BACKGROUND: Although there is a long history of the use spinal manipulation in China, little has been formally written on this topic. OBJECTIVE: To discuss the history and status of Chinese spinal manipulation. DISCUSSION: This article describes the history of Chinese spinal manipulation as revealed through past formal textbook publications on this topic, along with journal articles describing both the basic science foundation and clinical applications of such manipulation. In addition, the current status of Chinese spinal manipulation is described. CONCLUSION: There is a long history of the use of spinal manipulation throughout China, though some basic problems of application remain. Current research may help to resolve these problems. PMID- 9726066 TI - Chiropractic radiologists: a survey of demographics, abilities, educational attitudes and practice trends. AB - OBJECTIVE: To assess the education, abilities, practices and opinions of chiropractic radiologists. DESIGN: Eight-page survey with two mailings. PARTICIPANTS: One hundred seventy-five diplomates of the American Chiropractic Board of Radiology listed with the American Chiropractic College of Radiology office. RESULTS: Of the surveys mailed, 111 of the 175 were returned from the individuals listed with the American Chiropractic College of Radiology office. After excluding the deceased and those with a self-reported retired status, the response rate was calculated as 66%. Of those who responded, 81% were male; the mean age was 46.4 +/- 11.2 yr (mean +/- standard deviation). Most respondents were residents of California, followed by Canada and Illinois. Thirty-six percent taught at chiropractic colleges. They felt most confident in interpreting plain film studies of the musculoskeletal system. Many have undergone advanced training in specialized imaging. They felt topics related to patient positioning, plain film physics, arthritides, trauma, avascular necrosis and tumors are most important to a chiropractic curriculum. Most respondents felt the use of plain film radiology was most appropriate when the clinical presentation of the patient suggests the strong possibility of underlying disease. CONCLUSIONS: Radiology is an important topic to the education of students and the practice of chiropractic. The results of this survey will assist future decision-making regarding student education and clinical use of radiology. PMID- 9726068 TI - Strong and weak measures of efficacy: a comparison of chiropractic with biomedicine in the management of back pain. AB - A holistic, biocultural model for identifying efficacy in the management of back pain by chiropractic and medical doctors is proposed based on adopting the concepts of vertical reasoning and dual-level control. Using this approach, four different ways in which efficacy can be measured are identified: (a) anatomical physiological (curing of disease), (b) body/mind (healing of illness), (c) sociocultural (termination of sickness) and (d) political-economic (ending no access to care). These four are conceptualized as relating to levels in a hierarchy of structures of increasing size and complexity ranging from atoms and molecules to societies and nations. In measuring efficacy at each level, it is important to distinguish weak from strong measures. Strong measures conform to a Guttman scale model; each higher measure of efficacy also demonstrates efficacy at all levels lower than itself. Weak measures fail to conform to a Guttman-like scale, and are therefore of questionable value when used to support decisions relating to health care policy. PMID- 9726069 TI - Chiropractic management of a patient with subluxations, low back pain and epileptic seizures. AB - OBJECTIVE: To describe the chiropractic management of a patient presenting with complaints of low back pain and epileptic seizures. The discussion also addresses epilepsy and the current concepts of this disorder; possible mechanisms for the neurological effects of the chiropractic adjustment at sites of subluxation and its therapeutic implications are proposed. CLINICAL FEATURES: A 21-year-old woman with low back pain reported that she had fainted during the night and hit her head. She had been diagnosed since childhood with grand mal (tonicclonic) seizures as well as petit mal seizures. She had a seizure approximately every 3 hr, with a duration between 10 sec and 30 min for each episode. Examination indicated signs of subluxation/dysfunction at the L5-S1, C6-C7 and C3-C4 spinal levels. There was no evidence of cranial nerve involvement or any upper motor neuron lesion. Radiographic analysis revealed retrolisthesis of L5, hypolordosis of the cervical spine and hyperextension of the C6-C7 motion segment. INTERVENTION AND OUTCOME: Chiropractic adjustments using a specific-contact, short-lever arm, high-velocity, low-amplitude maneuver (i.e., Gonstead) were applied to the subluxations at the cervical, thoracic and lumbopelvic region. The patient's reported low back pain and neck complaints improved and her seizure frequency decreased. At 1.5-yr follow-up, the patient reported her low back complaints had resolved and her seizures had decreased (period between seizures as great as 2 months). CONCLUSION: Results encourage further investigation of possible neurological sequalae, such as epileptic seizures, from spinal dysfunction identified as vertebral subluxation complexes by chiropractors and treated by specific spinal adjustments. PMID- 9726070 TI - Spina bifida occulta mimicking a destructive lesion. AB - OBJECTIVE: To discuss an unusual presentation of spina bifida occulta (SBO) mimicking an aggressive bone lesion. CLINICAL FEATURES: A 59-yr-old man suffered from intermittent neck pain. The plain film radiographs revealed rarefaction of the C6 spinous process that mimicked an aggressive bone lesion. Computed tomography and previous radiographs of the region confirmed an SBO defect of the C6 level. INTERVENTION AND OUTCOME: Supportive therapy was applied to the patient for his presenting neck complaint. Chiropractic manipulative treatment was not applied to the lower cervical spine until the possibility of an aggressive bone lesion was excluded. CONCLUSION: An unusual presentation of SBO was presented. This case demonstrated the usefulness of specialized imaging to further delineate questionable anatomical presentations, which at times mimic destructive bone lesions. PMID- 9726071 TI - Nonspecific intervention in chiropractic care. PMID- 9726072 TI - Changes in brain function after manipulation of the cervical spine. PMID- 9726073 TI - Magnetic resonance imaging and clinical follow-up: study of 27 patients receiving chiropractic care for cervical and lumbar disc herniations. PMID- 9726074 TI - Most commonly used methods of detecting spinal subluxation and the preferred term for its description: a survey of chiropractic in Victoria, Australia. PMID- 9726075 TI - The effectiveness of chiropractic management of fibromyalgia patients: a pilot study. PMID- 9726076 TI - The effects of spinal manipulation on the intensity of emotional arousal in phobic subjects exposed to threat stimulus: a randomized, controlled, double blind clinical trial. PMID- 9726077 TI - The effects of spinal manipulation on the intensity of emotional arousal in phobic subjects exposed to threat stimulus: a randomized, controlled, double blind clinical trial. PMID- 9726078 TI - Rocking away the blues. PMID- 9726079 TI - Estrogen slows Parkinson's progression. PMID- 9726080 TI - A promising program for psychoeducation. PMID- 9726081 TI - Therapeutic holding: outcomes of a pilot study. AB - 1. Although decreasing the use of seclusion and restraints in the management of aggressive children is a critical issue facing pediatric psychiatric inpatient programs, finding effective alternatives has been a difficult challenge. 2. Therapeutic holding appears to be as effective as seclusion and restraint with respect to managing aggressive behaviors in the psychiatrically disordered child. 3. Therapeutic holding has the potential to reduce the episodes of mechanical restraints and to be perceived by children as less punitive. PMID- 9726082 TI - Wartime stressors and health outcomes: women in the Persian Gulf War. AB - This descriptive correlational study of war time stressors and stress responses of women from the Persian Gulf War examined numerous stressors both physical and psychological. The psychological stressors more directly impacted postwar physical and psychological symptoms than did physical stressors. These findings add to our understanding of women's reactions to wartime stress and the types of stressors affecting women. The study provides more data to support the contention that sexual harassment is widely prevalent in the military. The study did not find data to support concerns about maternal guilt on leaving children, nor any significant evidence of stress symptomology from this situation. The results of this study confirmed the call by Wolfe, Brown, Furey, and Levin (1993) for more precise evaluation of wartime stressors in view of the changing gender composition of military forces and the subsequent increase of women in combat roles. Clinicians should be alerted to recognize gender-specific experiences. Education of military women about stressors and coping mechanisms should be broadened to address the development issue of intimacy versus isolation. Nurses, both military and civilian, must understand the effect of isolation and discrimination on women both in combat and in other high stress situations. The need for continued study of the problem of sexual harassment is confirmed. Understanding the scope of the problem and the health care outcomes strengthens the role of prevention and intervention for nurses and their clients. PMID- 9726083 TI - Effectiveness of a counseling intervention to assist family caregivers of chronically ill relatives. AB - 1. Caregivers need information about community services, as well as ways to improve their ability to manage their relative's care. 2. Nurses can help ease a caregiver through the emotionally painful process of placing a relative in a nursing home by clarifying concerns and offering support and guidance. 3. Individual counseling is an effective intervention to assist family caregivers because the nurse can focus on the diverse needs and specific goals of each caregiver. PMID- 9726084 TI - Safer-sex education for persons with mental illness. AB - 1. Safer-sex education is crucial for controlling the spread of HIV and this education is within the realm of nursing practice. 2. Safer-sex education for persons with mental illness must include repetitive, interactive education capitalizing on verbal, visual, written, tactile, and motor skill teaching methods to compensate for learning disabilities that have been identified in this population. 3. In this study, individual rather than a group teaching approach to the sensitive subject of safer-sex provided the most learning. The study subjects did not exhibit overt sexual behavior, increased sexual acting out, regression, or overstimulation when safer-sex education was presented. PMID- 9726085 TI - An introduction to consumer politics. PMID- 9726086 TI - Irinotecan: toward clinical end points in drug development. AB - The objective response rate is the initial method to assess the activity of a novel anticancer agent. Response rates may not characterize a new agent's clinical benefit, however, especially if moderate to severe toxicity may be associated with treatment. Clinical end points, such as improvement in survival or relief of disease-related symptoms, provide clinicians with a more rational basis for selecting therapies. Irinotecan (CPT-11 [Camptosar]) was introduced into clinical practice based primarily on its consistent phase II activity (response rates of 13.3% to 21.7% in patients with advanced colorectal cancer refractory to 5-fluorouracil [5-FU]). Two recently presented, randomized, phase III trials have examined irinotecan's impact on clinical end points of survival, quality of life, tolerability, and control of disease-related symptoms in 5-FU refractory colorectal cancer patients. The first trial prospectively compared irinotecan and best supportive care (BSC) to BSC alone in patients with 5-FU resistant metastatic colorectal cancer. Irinotecan-treated patients had significantly prolonged survival, improved quality of life, and better control of disease-related symptoms. The second trial compared irinotecan to infusional 5-FU schedules in colorectal cancer patients whose tumors had progressed within 3 months of prior 5-FU. Patients treated with irinotecan lived significantly longer than those given infusional 5-FU and had a comparable quality of life. These randomized trials demonstrate an evolution in our understanding of the clinical utility of irinotecan. PMID- 9726087 TI - Rational design of irinotecan administration based on preclinical models. AB - Most clinical drug regimens for irinotecan (CPT-11 [Camptosar]) have been empirically based on classic in vivo pharmacokinetic and pharmacodynamic considerations. We propose an alternative approach that attempts to provide a rationally designed schedule of irinotecan administration based on preclinical data. HL60 cells grown in suspension or as subcutaneously implanted solid xenografts in nude mice served as in vitro and in vivo models to rest the activity of irinotecan or its active metabolite, SN-38. For SN-38, within an effective drug concentration range, scheduling drug administration based on duration of DNA synthesis inhibition significantly potentiated cell kill in vitro, and increasing drug concentrations at suboptimal scheduling did not result in additive cell kill. These data suggested that even though high drug doses may be attainable in vivo, they may not be required to achieve maximum antitumor activity. To test this hypothesis, a sensitive in vivo model to test the toxicity and antitumor activity of CPT-11 is required, which is provided in the human myeloid HL60 xenograft model grown in nude mice. In this model, CPT-11 at a dose 50 mg/kg, daily x5 (MTD) achieved 100% complete tumor regression. This model should be useful to test the hypothesis that for irinotecan, administration of a minimum effective dose (MED) at an optimal schedule can achieve maximum antitumor activity and should therefore prevail over the classic approach of administering the MTD. PMID- 9726088 TI - Irinotecan: a review of the initial phase I trials. AB - The unique mechanism of action of irinotecan (CPT-11 [Camptosar]), topoisomerase I inhibition, together with the results of preclinical studies, suggest that the drug's antitumor and toxicologic effects may be schedule-dependent. To further explore this possibility, we reviewed the initial phase I studies of various administration schedules that have been conducted in Japan, France, and the United States. This review showed toxicities to be fairly consistent across dosing schedules, although the severity and extent of diarrhea and neutropenia differed somewhat. The institution of intensive loperamide therapy and perhaps myeloid growth factors may have allowed for further dose escalation on some schedules, although it is unclear whether dosing intensity should be pursued without regard to dosing frequency. Preliminary antitumor activity of irinotecan noted in a study of leukemia and lymphoma supports the theory that the drug may exhibit schedule-dependent antitumor activity. The results of these early studies of irinotecan should be taken into account when designing subsequent trials of the agent alone or in combination with other chemotherapeutics in specific tumor types. PMID- 9726089 TI - Pharmacology of irinotecan. AB - Irinotecan (CPT-11 [Camptosar]), a semisynthetic derivative of the plant alkaloid camptothecin, is bioactivated by carboxylesterases (EC3.1.1-) to the topoisomerase I inhibitor SN-38, a minor metabolite. Bioactivation of intravenously administered irinotecan by carboxylesterases occurs predominantly in the liver. Two human carboxylesterase isoforms responsible for SN-38 formation have been characterized. At relevant hepatic irinotecan concentrations up to 12 micrograms/mL, a low-Km isoform is responsible for irinotecan bioactivation. High concentrations of drugs commonly coadministered with irinotecan do not inhibit carboxylesterase activity. Intestinal carboxylesterases can also generate SN-38, followed by subsequent oral absorption. A second major polar metabolite of irinotecan, aminopentanecarboxylic acid (APC), is the product of CYP3A4-mediated oxidation of the terminal piperidine ring. APC is 100-fold less active than SN-38 as a topoisomerase I inhibitor and is a relatively weak inhibitor of acetylcholinesterase. SN-38 is eliminated mainly through conjugation by hepatic uridine glucuronosyltransferase (UGT*1.1), the same isoezyme responsible for glucuronidation of bilirubin. Grade 4 irinotecan-related toxicity (ie, neutropenia, diarrhea) has recently been reported in two patients with deficient UGT*1.1 activity. SN-38 glucuronide (SN-38G), which has only 1/100th the antitumor activity of SN-38, is actively secreted into the bile by a canalicular multispecific organic anion transporter. Deconjugation of SN-38G to SN-38 by beta glucuronidase produced by the intestinal flora may contribute to enterohepatic recirculation of SN-38 and delayed intestinal toxicity. PMID- 9726090 TI - Thymidylate synthase as a predictor of response. AB - It has been hypothesized that intratumoral thymidylate synthase (TS) gene expression might be used to select therapy for patients with disseminated colorectal cancer. We recently reported the results of a clinical trial in 46 patients with disseminated or recurrent colorectal cancer testing whether expression of TS within the primary tumor, as assessed by quantitative polymerase chain reaction (PCR) methodology, would predict the responsiveness of that cancer of fluoropyrimidine-based therapy. This trial demonstrated that intratumoral TS/beta-actin messenger RNA (mRNA) ratio can accurately predict which metastatic colorectal tumors will be resistant to a leucovorin-modulated 5-FU infusion and which have a high likelihood of responding to such a regimen. Results of other studies of adjuvant therapy in gastric cancer and colorectal cancer also indicated that TS expression within the tumor is predictive of response of 5-FU based therapy. It may be possible to use this parameter prospectively to decide which patients should receive fluorinated pyrimidine therapy: Patients whose tumors express low TS levels would be likely to benefit from such therapy, whereas limited preliminary data suggest that patients whose tumors express high TS levels may benefit from irinotecan (CPT-11[Camptosar]). PMID- 9726091 TI - US pivotal studies of irinotecan in colorectal carcinoma. AB - Phase I trials of irinotecan (CPT-11 [Camptosar]), conducted at Johns Hopkins and the University of Texas, San Antonio, demonstrated some activity in patients with refractory advanced cancer. Three pivotal phase II studies of irinotecan in advanced colorectal carcinoma were conducted at The University of Texas, San Antonio, Mayo/North Central Cancer Treatment Group (NCCTG), and the CPT-11 Study Group in a total of 304 patients. All patients had received prior fluorouracil (5 FU) chemotherapy, and over 90% had progressed while on treatment within the last 6 months. The initial starting dose of irinotecan ranged from 100 to 150 mg/m2. The overall response rate was 12.8% (95% confidence interval, 9.1% to 16.6%) with a 15% response rate at a recommended starting dose of 125 mg/m2. The response durations and overall median survivals were similar in the three studies. The principal toxicities included diarrhea, nausea, vomiting, and neutropenia. Severe diarrhea was limited by use of an intensive loperamide regimen and appropriate dose modification. The three pivotal studies of irinotecan in advanced colorectal carcinoma demonstrate consistent response rates and duration, with manageable toxicity. Future studies will focus on the use of irinotecan in chemotherapeutically naive colorectal carcinoma, the adjuvant treatment of colon carcinoma, combination chemotherapeutic regimens, and treatment of other malignant diseases. PMID- 9726092 TI - Irinotecan in the first-line treatment of colorectal cancer. AB - Irinotecan (CPT-11 [Camptosar]) is currently approved for use as a second-line agent in the treatment of metastatic colorectal cancer. Phase II studies have also shown substantial single-agent activity of irinotecan in the first-line treatment of metastatic colorectal cancer. Response rates appear to be similar to those seen with standard first-line regimens, although direct randomized comparisons have not yet been reported. In the absence of definitive data showing irinotecan to be superior, its routine use as a single agent in the first-line treatment of colorectal cancer may be hard to justify, given the significant cost differential between irinotecan and current standard first-line regimens. Studies exploring combinations of irinotecan with fluorouracil may identify a first-line role for these combination regimens. Also, use of specific molecular markers may permit the identification of selected patients with tumor characteristics that would specifically favor consideration of upfront irinotecan monotherapy. PMID- 9726093 TI - Irinotecan plus 5-FU and leucovorin in advanced colorectal cancer: North American trials. AB - But fluorouracil (5-FU) and irinotecan (CPT-11 [Camptosar]) have shown activity in metastatic colorectal cancer and are approved for its treatment in the United States. Preclinical experiments in cell cultures and human tumor xenografts have indicated potential synergy when irinotecan is combined with 5-FU and leucovorin. The synergy appears to be sequence-dependent and is optimal when irinotecan exposure precedes 5-FU exposure by at least 24 hours. Four North American trials have been reported in which the three drugs were used together in either simultaneous, sequential, or alternating schedules. All three schedules showed activity in patients with metastatic colorectal cancer. The concern that diarrhea, which can be a dose-limiting toxicity with both irinotecan and 5-FU, would prevent the two drugs from being combined in reasonable doses has not proven to be a clinical issue. Phase III trials comparing the combination of the three drugs in a variety of schedules against 5-FU plus leucovorin alone are currently under way or in the planning stages. PMID- 9726094 TI - European experience with irinotecan plus fluorouracil/folinic acid or mitomycin. AB - Tremendous progress has been made in the medical treatment of advanced colorectal cancer during the past 2 to 3 years, due to the availability of several new drugs. Of these new agents, irinotecan (CPT-11 [Camptosar]) seems to be one of the most active against advanced colorectal cancer. It is, therefore, a good candidate for combination with the more classic cytotoxic agents for this disease. This article summarizes several European phase I and II studies in which irinotecan has been combined with (1) fluorouracil (5-FU) alone, given as a repeated bolus injection or a protracted infusion; (2) 5-FU modulated by folinic acid (leucovorin) according to different schedules; or (3) mitomycin (Mutamycin). All of these studies have demonstrated clinical responses in patients with advanced colorectal carcinoma, including complete responses. The toxicity profiles of the various combinations seem to be acceptable; neutropenia and delayed diarrhea are the most frequent side effects. Large phase III studies are still warranted to demonstrate the benefit of these irinotecan-based regimens. PMID- 9726095 TI - Alternative dosing schedules for irinotecan. AB - Most of the clinical experience with irinotecan (CPT-11 [Camptosar]) has been with either a weekly or an every-3-week schedule. Recent phase I trials have explored new routes and schedules of administration. One approach attempts to maximize dose frequency and intensity by giving irinotecan every 2 weeks. A phase I trial of this approach is now complete and has led to a phase II trial in patients with recurrent colorectal cancer. Data suggest that smaller doses of a topoisomerase I inhibitor administered repeatedly may result in greater antitumor activity than large doses administered intermittently. A phase I trial has been performed in adults in which irinotecan was administered daily for 5 consecutive days, followed by 2 days off, for 2 weeks out of 3. Similar trials are under way in children. Oral administration, another strategy that has undergone phase I testing, has several theoretical advantages:(1) The acidic pH of the stomach favors maintenance of irinotecan in the active lactone ring form. (2) Irinotecan is more rapidly and extensively converted to SN-38 by tissue carboxylesterases found in high concentrations in the gut and liver. (3) Low doses can be delivered over a protracted period. (4) The oral route enhances patient convenience. These alternative dosing schedules may facilitate integration of irinotecan into combination chemotherapy and combined-modality treatment regimens. PMID- 9726096 TI - Gastrointestinal toxicity or irinotecan. AB - Irinotecan (CPT-11 [Camptosar]) is an important new chemotherapeutic drug that demonstrates activity against a broad spectrum of malignancies, including carcinomas of the colon, stomach, and lung. Unfortunately, frequent and often severe gastrointestinal toxicities, particularly diarrhea, have limited its more widespread use. A cholinergic syndrome resulting from the inhibition of acetylcholinesterase activity by irinotecan is frequently seen within the first 24 hours after irinotecan administration but is easily controlled with atropine. Late diarrhea occurs in the majority of patients, however, and is National Cancer Institute (NCI) grade 3 or 4 in up to 40%. The late syndrome appears to be related to the effects on the bowel of SN-38, the active metabolite of irinotecan, which undergoes biliary excretion and inactivation. Early recognition and treatment of late diarrhea with high-dose loperamide have reduced, although not entirely eliminated, patient morbidity. Further study is needed to identify the mechanism of irinotecan-induced late diarrhea and to evaluate potential new therapies. PMID- 9726097 TI - Irinotecan plus cisplatin in patients with advanced non-small-cell lung cancer. AB - During the 1980s, platinum-based regimens were yielding response rates typically less than 25%, median survival durations of about 25 weeks, and 1-year survival rates less than 25% in patients with advanced non-small-cell lung cancer (NSCLC). Currently, results from single institution phase II trials of agents introduced in the 1990s show a doubling of these numbers, and results from multiinstitutional trials are demonstrating response rates ranging from 30% to 40%, median survival durations of 40 weeks, and 1 year survivals of 40%. Single agent irinotecan shows significant activity against NSCLC in preclinical and early phase I/II clinical studies, with activity similar to that for other new agents. Therapeutic synergy is observed in preclinical tumor models when irinotecan and cisplatin are combined, and phase I/II trials of this combination have demonstrated response rates > or = 50%. Herein the author provides an overview of data from phase II trials of irinotecan and focuses on preliminary results of a large US multicenter phase II trial of weekly irinotecan plus monthly cisplatin in 52 patients with advanced NSCLC. A response rate of 28.9% (95% CI, 16.5%-41.2%) and a median survival of 9.9 months were observed in this trial. US studies to design a more optimal irinotecan/cisplatin regimen in the same patient population are ongoing, and early results are encouraging. PMID- 9726098 TI - Extending principles learned in model systems to clinical trials design. AB - Clinical results with irinotecan (CPT-11 [Camptosar]) and other camptothecin derivatives in various cancers, although encouraging, have fallen short of the expectations predicted by preclinical models. One proposed explanation for this is that preclinical xenograft models do not predict for the sensitivity of human cancer. In this article, we describe the results of several studies suggesting that this explanation is incorrect. Instead, our results indicate that the discrepancy between clinical response rates and findings in preclinical models may be due to a failure to incorporate the principles learned from preclinical studies into the design of clinical trials. Our analysis suggests that if differences in host tolerance are taken into account, the xenograft models are quite accurate predictors of clinical response. Moreover, application of the principles derived from preclinical models to the design of clinical trials may significantly enhance clinical response rates. Thus, the camptothecin analogs provide a paradigm for better integrated, pharmacokinetically driven, preclinical and clinical development of new drugs. PMID- 9726099 TI - Irinotecan in cervical cancer. AB - Several studies have evaluated the use of irinotecan (CPT-11 [Camptosar]), a topoisomerase inhibitor, in the treatment of refractory or recurrent cervical cancer. Various schedules have been used. Response rates have ranged from 13% to 20%. One phase I study of the combination of cisplatin (Platinol) and irinotecan has been completed. Toxicities have been hematologic and gastrointestinal. The latter remains problematic. Further studies of irinotecan in combination with other drugs, particularly cisplatin, are recommended. PMID- 9726100 TI - Current status of therapy for advanced gastric carcinoma. AB - Advanced gastric carcinoma remains an incurable disease with a median survival of 6 to 9 months, and available therapeutic approaches are predominantly palliative. In small controlled trials, systemic chemotherapy has improved survival and quality of life of patients with advanced gastric carcinoma when compared with best supportive care. Patients with good performance status (Zubrod < or = 2), low tumor bulk, and good organ function are most likely to benefit from chemotherapy or combined-modality therapy. There is no generally accepted standard chemotherapy for advanced gastric carcinoma. Fluorouracil-and/or cisplatin-based combinations are often employed. Recently, several new classes of drugs have demonstrated activity against advanced disease. These include the taxanes (paclitaxel [Taxol] and docetaxel [Taxotere]), camptothecins (irinotecan [Camptosar], and flurouracil prodrugs (second-and third-generation agents, such as UFT [uracil and tegafur] and S-1). Early results with either single-agent therapy or combinations of new agents (irinotecan, paclitaxel, and docetaxel) and more conventional agents (cisplatin [Platinol] and fluorouracil) are encouraging. Several of these results need to be confirmed and eventually studied in well designed, phase III trials. Similarly, a number of new combinations may be used in the future as preoperative therapies for gastric carcinoma. Nearly all of the new agents have radiosensitizing properties. This affords another opportunity to investigate new chemotherapeutic agents in conjunction with radiation therapy in patients with locoregional gastric carcinoma. PMID- 9726101 TI - Irinotecan in lymphoma, leukemia, and breast, pancreatic, ovarian, and small-cell lung cancers. AB - Irincotecan (CPT-11 [Camptosar] has a board range of antitumor activity. Extensive preclinical and early clinical work has demonstrated its activity against many tumor types--head and neck, esophagus, stomach, pancreas, liver, colon/rectum, kidney, lymph nodes, ovary, uterine, cervix, sarcoma, melanoma, acute and chronic leukemia, mesothelioma, and cancers of unknown primary site. Most of the phase II and III trials have focused on colorectal and other gastrointestinal, non-small-cell lung, and cervical cancers (discussed elsewhere in this monograph). This article presents preliminary results of studies exploring the use of irinotecan in lymphoma, leukemia, and breast, pancreatic, ovarian, and small-cell lung cancers. In all of these studies, the number of patients enrolled is small, drug doses and schedules differ (often within the same case series), and little information is available on response duration and overall survival. Nevertheless, irinotecan has shown reproducible if at times modest activity in almost all of the diseases in which it has been studied. Future research should be directed at conducting well-designed clinical trials of irinotecan alone and in combination with other agents. PMID- 9726102 TI - Irinotecan and cisplatin in upper gastrointestinal malignancies. AB - Irinotecan (CPT-11 [Camptosar]), an active agent in the treatment of fluorouacil refractory colorectal cancer, has antitumor activity in upper gastrointestinal cancers. Clinical trials from Japan indicate antitumor responses in gastric and pancreatic cancers. Cisplatin (Platinol), a central agent in the treatment of upper gastrointestinal malignancies, is a logical drug to study in combination with irinotecan in upper gastrointestinal cancers. In vitro studies have shown important sequence-dependent synergy of cisplatin/irinotecan combination therapy. Irinotecan appears to prevent removal of cisplatin-induced DNA-interstrand cross links. Initial phase I and III trials of cisplatin plus irinotecan appear to confirm this synergy, with Japanese trials in gastric cancer showing an encouraging rate of response with acceptable toxicity. A phase I trial conducted at Memorial Sloan-Kettering Cancer Center has demonstrated the safety and tolerability of weekly cisplatin and irinotecan. Currently, a phase II trial of this weekly regimen is under way in patients with metastatic or recurrent esophageal cancer. The response proportion compares favorably to standard therapy, with relatively mild toxicity. Other phase II studies, including single agent irinotecan in esophageal cancer and the combination of cisplatin and irinotecan in gastric cancer, are being initiated at other US institutions. PMID- 9726103 TI - Camptothecin radiation sensitization: mechanisms, schedules, and timing. AB - Based on high tumoricidal activity of the camptothecin analogs topotecan (Hycamtin), irinotecan (CPT-11[Camptosar]), and 9-aminocamptothecin (9-AC) in preclinical studies, clinical trials began testing these agents against human cancers. The cytotoxic activity of camptothecins in the clinic has been lower than predicted from the laboratory, however, and new approaches are needed. One method that holds promise is the use of the camptothecins as radiation sensitizers. The camptothecin dose, schedule, method of administration, and timing of administration, when given with irradiation, are likely to be important factors for these new S-phase radiation sensitizers. Phase I trials of the camptothecins as radiation sensitizers have begun, and multicenter phase II studies are planned by the Radiation Therapy Oncology Group (RTOG). One new approach based on preclinical observations that deserves clinical evaluation is chronomodulated camptothecin delivery with irradiation in order to widen the therapeutic window. PMID- 9726104 TI - Assessing the impact of chemotherapy on tumor-related symptoms in advanced colorectal cancer. AB - In all patients with advanced colorectal cancer, disease eventually progresses following fluorouracil (5-FU) therapy, with a worsening of disease-related symptoms and quality of life (QOL). Irinotecan (CPT-11 [Camptosar]) has produced response rates of 16% to 27% in patients with 5-FU-refractory colorectal cancer, along with a modest survival gain and possible palliative benefit. Disease related symptoms and QOL are major end points of palliation, although few studies have assessed them as primary end points of response. The concept of palliative response benefit has been applied successfully to randomized trials of systemic therapy in prostate and pancreatic cancers. This article will describe how this concept has been used in the design of a current phase II trial assessing the palliative benefit of irinotecan in patients with 5-FU-refractory colorectal cancer. This trial uses disease-related symptoms and performance status as primary end points. Palliative response was defined prospectively as one or more of the following; (1) 50% decrease in pain score; (2) 50% decrease in narcotic analgesic usage; or (3) 10-point increase in Karnofsky performance scale from baseline for 4 weeks, without deterioration of any of these parameters. The difficulties in using patient-oriented end points as response criteria in this trial and in the clinic will be addressed. PMID- 9726112 TI - Biculturalism and perceived competence of Latino immigrant adolescents. AB - The present study investigated acculturation to the Hispanic and American cultures and self-perceptions of competence among 123 Latino immigrant adolescents. The study tested a contextual model of biculturalism by examining whether different acculturation styles predicted perceived competence in life spheres with different cultural demands. Perceived competence was assessed using Harter's (1988). Self-Perceptions of Competence Profile for Adolescents for the life spheres of school, peers (both Latino and non-Latino), and global self worth. In addition, an analogous scale to assess perceptions of competence in the family was constructed for that sphere. The study found some support for a contextual model of acculturation. Acculturation to American culture predicted positive self-perceptions of competence with American peers, while acculturation to Hispanic culture predicted positive self-perceptions of competence with Latino peers. Perceived family competence, however, was predicted by acculturation to American rather than Hispanic culture. Results with respect to biculturalism are tentative, with a trend relating biculturalism to positive self-perceptions of global self-worth. However, because many of the conditions stipulated by the model were not met, results with respect to biculturalism raise questions about current approaches to operationalizing the construct. PMID- 9726113 TI - The community response to rape: victims' experiences with the legal, medical, and mental health systems. AB - This research examined how the legal, medical, and mental health systems respond to the needs of rape victims. A national random sample of rape victim advocates (N = 168) participated in a phone interview that assessed the resources available to victims in their communities, as well as the specific experiences of the most recent rape victim with which they had completed work. Results from hierarchical and iterative cluster analysis revealed three patterns in victims' experiences with the legal, medical, and mental health systems. One group of victims had relatively positive experiences with all three systems, a second group had beneficial outcomes with only the medical systems, and the final group had difficult encounters with all three systems. Multinominal logistic regression was then used to evaluate an ecological model predicting cluster membership. Community-level factors as well as features of the assault and characteristics of the victims predicted unique variance in victims' outcomes with the legal, medical, and mental health systems. These findings provide empirical support for a basic tenet of ecological theory: environmental structures and practices influence individual outcomes. Implications for ecological theory and interventions to improve the community response to rape victims' needs are discussed. PMID- 9726114 TI - Academic achievement of African American preadolescents: the influence of teacher perceptions. AB - The purpose of this study was to identify the factors associated with the academic success of predominantly, middle-class African American preadolescent students. This study proposed an ecological model that considered the interaction of family environment, teacher perceptions of social skills, and student characteristics. The estimated model explained 58% of the variance in grade point average. Path analysis revealed three direct effects on grade point average, (a) grade level (negative), (b) teacher perceptions of social skills, and (c) academic ability. Findings revealed that teacher perceptions of social skills was a stronger predictor of grade point average than academic ability. Two indirect effects on grade point average were found. The first indirect effect was negative: gender predicted academic ability, which predicted teacher perceptions of social skills, which predicted grade point average. The second indirect effect was positive and it was from ability to teacher perceptions to grade point average. Implications for policy and practice are made that suggest a collaborative model of school counseling designed to "promote" the social and academic competence of African American students. Interventions that enhance teacher practices are also suggested. PMID- 9726115 TI - An ecologically differentiated, multifactor model of adolescent network orientation. AB - The paper presents a test of an ecologically differentiated model of social network orientation for adolescents that distinguished between different social network reference groups (family, peers, and nonfamily adults). The model was tested in two consecutive studies. Study 1 describes initial model development (N = 120). Study 2 presents a confirmatory factor analysis with a second sample (N = 430) to replicate the factor structure developed in Study 1. Results supported a three-factor model of network orientation that differentiated between network reference groups. Analyses of concurrent and predictive validity indicated that orientation to network reference groups was differentially related to the perceived quality and frequency of support from members of respective social network groups. Group differences (gender, race) regarding network orientation to different network reference groups were consistent with studies of other social network processes. Implications for the study of the network orientation and the study of social networks more generally are discussed. PMID- 9726116 TI - Alcohol and drug use, and depression among Hispanic men in early adulthood. AB - Community research and clinical practice have shown that alcohol and drug use and depression are interrelated. Among Hispanics, acculturation may play a role in these relationships. To investigate these relationships as well as alcohol related problems, we interviewed 288 Puerto Rican, Dominican, and Colombian men in early adulthood. No significant differences emerged in the proportions of abstainers across the three groups. Colombians drank significantly more frequently and had more alcohol-related problems than Dominicans. Dominicans were at least risk for and least likely to have alcohol-related problems. Puerto Ricans were much more likely to use drugs than the other Hispanic men. Drug use was associated with an increased likelihood of heavy drinking which, in turn, increased the risk of drug use and depression. Acculturation decreased the risk of drug use. Results are discussed in terms of implications for community psychology research and interventions. PMID- 9726117 TI - Partners and fellow patients: two sources of emotional support for women with breast cancer. AB - This study examined the helping process that occurred when 26 breast cancer patients (the disclosers) talked about their illness-related concerns with their partner and, in a separate conversation, with a fellow patient (the volunteer helpers). The conversations were rated by trained observers and by the disclosers in terms of several process and outcome variables. From the observers' perspective, the volunteer helpers were more helpful, empathic, and supportive, less critical, and used more self-disclosure than the partners. Disclosers did not differentiate between the two types of helper, and gave generally high ratings to both conversations. Strengths and weaknesses of each type of helper were identified. Findings are discussed in relation to the literature on formal and informal helping, and implications for training nonprofessional helpers are suggested. PMID- 9726118 TI - Remaining radical? Organizational predictors of rape crisis centers' social change initiatives. AB - Rape crisis centers have undergone significant changes since their birth during the feminist movement of the 1970s. As has happened with many other radical social movements, there is growing evidence that the antirape movement has become more institutionalized. This research used a combination of quantitative and qualitative methods to examine the current structure and functions of a national random sample of 168 rape crisis centers. An organizational-level model predicting involvement in three types of social change activities was tested: (a) participation in public demonstrations to raise awareness about sexual assault; (b) political lobbying for violence against women legislation; and (c) primary prevention programs to eliminate sexual violence against women. Results of logit modeling suggested that how long a rape crisis center had been in existence moderated the relationships between organizational characteristics and involvement in community activism. Findings of this study suggest that although many of today's centers bear little resemblance to the grass-roots collectives of years past, rape crisis centers have been remarkably adaptive in weathering changing political climates to continue to provide comprehensive services for rape victims. PMID- 9726119 TI - Ultrastructural characterization of atrial natriuretic peptide receptors (ANP-R) mRNA expression in rat kidney cortex. AB - Atrial natriuretic peptide (ANP) and two complementary peptides named brain natriuretic peptide and C-type natriuretic peptide are involved in diuresis, natriuresis, hypotension and vasorelaxation. Their actions are mediated by highly selective and specific ANP receptors. Three subtypes have been characterized and cloned: ANP receptor A, -B and -C. In the present study, the mRNA for each subtype was detected by ultrastructural in situ hybridization on ultrathin sections of Lowicryl-embedded tissue and frozen tissue. The distribution of mRNA (visualized by gold particles) for each subtype was found to differ in different cells of the nephron. The three subtypes of this receptor family were expressed in all the parts of the nephron, but their expression levels were different. The ANPR-A mRNA was the most abundant in cells of glomerulus, proximal and distal tubules. The subtype C was the least expressed mRNA in glomerulus. In contrast, the subcellular localization of the three mRNAs was similar; they were found in the cytoplasmic matrix and the euchromatin of the nucleus. In conclusion, the differential expression of these mRNAs in kidney cortex indicates that these three peptides act directly in differing parts of nephron regions which are the glomerulus, the proximal and distal tubules. PMID- 9726120 TI - Atomic force microscopy and modeling of natural elastic fibrillin polymers. AB - A central issue in the understanding of Marfan syndrome deals with the functional architecture of fibrillin-containing microfibrils. Fibrillin-rich microfibrils are long extracellular matrix fibrillar components exhibiting a 50 nm periodic beaded-structure with a width of around 20-25 nm after rotary shadowing and a 10 12 nm diameter when observed in ultra-thin sections. They are composed of fibrillin monomers more or less associated with many other components which are, for the most part, poorly characterized up to date. They are known to be elastic but few data have been accumulated to understand their properties. Atomic force microscopy (AFM) allowed us to morphologically differentiate fibrillin-rich microfibrils from other fibrillar components and to investigate the thin structure of these beaded filaments in their native state. They showed, in AFM, a periodic beaded structure ranging from 50 to 60 nm and a width of about 40 nm. The different sizes of fibrillin-containing microfibrils previously observed after rotary shadowing and in ultra-thin sections was resolved with our technique and is revealed to be 10 nm in diameter. Each beaded microfibril appears to be composed of heterogeneous beads connected by 2-3 arms. An orientation of the microfibrils has been shown, and allows us to propose a complementary model of microfibrillar monomer association. PMID- 9726121 TI - Lysosomal involvement in the removal of clofibrate-induced rat liver peroxisomes. A biochemical and morphological analysis. AB - Peroxisomal proliferators induce in rodents hepatic hyperplasia and hypertrophy; the significant increase in the peroxisomal population is accompanied by specific and reversible induction of some peroxisomal enzymes. In suckling rats born from clofibrate-treated mothers, a massive removal of proliferated organelles occurs within 3 days of recovery. In the present paper we examined the early stages of the recovery period in liver of male rats treated with clofibrate for 5 days. The lysosomal involvement in the removal of drug-induced peroxisomes was investigated under physiological conditions, i.e. in the absence of inhibitors of the autophagic process. Biochemical results indicate that peroxisomal beta-oxidation, but not catalase activity, returns to the control values within the examined period. Total acid phosphatase activity is not affected by clofibrate treatment, but following fractionation on a linear density gradient the lysosomal marker enzyme activity is shifted towards lower density values, particularly at day 1 and 2 of recovery. This class of organelles possibly represents lysosomes involved in active autophagic processes. Acid phosphatase cytochemistry shows an increase of lysosome number at day 1 of recovery. Combination of acid phosphatase cytochemistry either with catalase cytochemistry or with catalase immunogold labelling allows to reveal organelles containing both marker enzymes. These results strongly support the involvement of autophagic processes in the removal of proliferated peroxisomes. PMID- 9726122 TI - Leishmania major: cell type dependent distribution of a 43 kDa antigen related to silent information regulatory-2 protein family. AB - In previous studies we have characterized several Leishmania major polypeptides and showed that one member of this group (LmSIR2rp) shared significant homology to silent information regulator 2 (SIR2) of Saccharomyces cerevisiae, a protein playing a role in both telomeric and mating type loci repression in these organisms. In the present study, by using molecular and immunological approaches, we could identify LmSIR2rp homologues in different Leishmania species and developmental stages (e.g. logarithmic (LP) and stationary phase promastigotes (SP) and amastigotes). The reactive antigen was also detected in Trypanosoma cruzi extracts. Surprisingly, immunofluorescence assays revealed that LmSIR2rp is associated mainly with cytoplasmic granules of different sizes and numbers depending on the life stage of the parasite used. No reactivity was observed in the nucleus, in agreement with the Western blot showing an absence of immunoreactivity of anti-LmSIR2rp immune serum against parasite nuclear extracts. Furthermore, immunoprecipitation of [35S]methionine-labeled promastigote antigens after pulse chase experiments, using anti-LmSIR2rp fusion protein antibodies, showed that the protein is among parasite excreted-secreted antigens (ESA). Moreover, immunofluorescence assays conducted with short time incubations of either purified LmSIR2rp or viable promastigotes with murine macrophages, revealed that LmSIR2rp could be bound to the macrophage surface. The unexpected cytoplasmic localization of LmSIR2rp and its presence in ESA may suggest a new mode of action for silent information regulatory factor homologues. PMID- 9726123 TI - Regulation of pectic enzymes from the exo-1 mutant strain of Neurospora crassa: effects of glucose, galactose, and galacturonic acid. AB - The exo-1 mutant of Neurospora crassa produced and secreted pectolytic activities when incubated in the presence of pectin-containing biological materials. This study shows that polygalacturonase, pectate lyase and pectin lyase activities were induced in media supplemented with galactose or galacturonic acid, indicating that these sugars induced the synthesis of pectinases. Pectinesterase activity was undetectable. Polygalacturonase activity was better induced by galactose than by galacturonic acid. The reverse was true for lyase activities. The inducing effect of galactose and galacturonic acid seemed to be different: (i) a mixture of galactose and galacturonic acid synergistically increased the production of pectic enzymes, as compared to that in the presence of one of these sugars; (ii) the inducing effect of galacturonic acid was partially repressed by glucose; (iii) in contrast, the inducing effect of galactose, rather than repressed, was enhanced by the presence of glucose. Altogether, these data point out to a complex mechanism of regulation of pectolytic enzymes by pectin containing organic substances. PMID- 9726124 TI - Purification and properties of methanol dehydrogenase from Methylosinus sp. WI 14. AB - Similarly to the recently described methanol dehydrogenase (MDH) from Methylocystis sp. GB 25 (Grosse et al. 1997) MDH from Methylosinus sp. WI 14 is able to catalyse the oxidation of methanol to formate directly. The enzyme was purified about 9-fold to electrophoretic homogeneity and is localised in the soluble fraction. The relative molecular mass of the native enzyme has been determined to be 140 kDa. It is composed of two identical subunits of relative molecular mass 70 kDa. The amino terminal sequence shows a strong similarity (a match of 80% over the first 20 amino acids) to the MDH from Methylocystis sp. GB 25. PQQ could be detected as the prosthetic group of MDH in the purified enzyme fraction by using the apoenzyme of a membrane-bound glucose dehydrogenase from Pseudomonas aeruginosa. A PQQ ratio of 1.3 per mole MDH was estimated. The purified enzyme has an optimum activity at pH 9.0 and at 57 degrees C. MDH from Methylosinus sp. WI 14 oxidises only primary alcohols up to octanol and several aldehydes. The estimated K(m)-values vary between 0.18 mM for the sorbic alcohol and 6.3 mM for butanol and show no dependence upon the chain length. PMID- 9726126 TI - Postantibiotic effect of some antibiotics on the metabolism of Pseudomonas aeruginosa. AB - The influence of the postantibiotic effects (PAEs) of ciprofloxacin, pefloxacin, imipenem, meropenem and amikacin in the suprainhibitory concentrations (2 x and 4 x MIC) on the metabolic processes of P. aeruginosa was studied. The synthesis of macromolecules was expressed by influencing of the incorporation rate of [14C] adenine and [14C] leucine. Remarkable affecting of both biosynthetic processes evoked the suprainhibitory concentration 4 x MIC of meropenem by inhibition of the nucleic acids synthesis to 76.1% and proteins synthesis to 61.1% against the control. The suprainhibitory concentration 4 x MIC of both pefloxacin and ciprofloxacin affected the highest suppression of the endogenous respiration to 16.5% and to 20.3%, respectively. The respiration was influenced the least after the effect of meropenem in the both suprainhibitory concentrations tested. According to our knowledge, this is first report about the evaluation of the endogenous respiration after PAE. In this study we demonstrated the inhibitory effects of 4 x MIC concentration of antibiotics studied on the metabolic processes of P. aeruginosa. The results suggest a multiple mechanism for the PAE. PMID- 9726125 TI - Key enzymes for the degradation of benzoate, m- and p-hydroxybenzoate by some members of the order Actinomycetales. AB - A preliminary screening of numerous species of the order Actinomycetales, especially of the genera Mycobacterium, Nocardia, Rhodococcus, Pseudonocardia, and Streptomyces, showed that many of them are able to metabolize benzoate (B) and p-hydroxybenzoate (pHB) as indicated by growth and change of color of the pH indicator of an agar medium. Subsequent experiments with liquid cultures which allowed the analysis of substrate utilization by thin layer chromatography confirmed these results. The study of the degradative pathway proved that B was metabolized via catechol (C), pHB via protocatechuate (P) and m-hydroxybenzoate (mHB) via gentisate (G). The aromatic ring of C and P was subjected to an ortho cleavage; only one strain of Noc. asteroides degraded C via a meta-cleavage, but P via an ortho-cleavage. Cell free extracts of four selected organisms exhibited activity of C-1,2-dioxygenase (C-1,2-O) and/or P-3,4-dioxygenase (P-3,4-O), depending on the growth substrate used for precultivation. In Streptomyces C-1,2 O was only found in cells grown on B, and P-3,4-O only in cells grown on pHB. On the contrary, in Rhodococcus rhodochrous B-cells oxidized C as well as P, while P cells possessed only P-3,4-O-activity. PMID- 9726127 TI - Medicaid estate planning: practices and perceptions of Medicaid workers, elder law attorneys, and certified financial planners. AB - This study examined Medicaid estate planning (MEP) through the experiences and perceptions of three groups in Connecticut: Medicaid eligibility workers (n = 128), elder law attorneys (n = 41), and certified financial planners (n = 29). Respondent groups varied significantly with regard to their perceptions of prevalence and magnitude of MEP, the nature of transferred assets, mechanisms for transfers, and characteristics of the "typical" client participating in asset divestiture for the purpose of qualifying for Medicaid. This substantial lack of concordance among those professionals most closely involved with MEP poses challenges for policy and research in this area. PMID- 9726129 TI - Everyday competence in later life: current status and future directions. AB - This article reviews the literature on the antecedents and outcomes of everyday competence in later life and discusses future directions. It is argued that there is a fairly solid knowledge base with regard to the antecedents and outcomes but not in terms of the components and mechanisms of older adults' everyday competence. Five key issues are identified and discussed in terms of a future research agenda. For each key issue, directions for future research are outlined and a transactional approach is advocated. It is emphasized that older adults should be viewed as proactive individuals who are motivated to minimize the losses and maximize the gains associated with the aging process. PMID- 9726128 TI - Home-based resistance training: predictors of participation and adherence. AB - This study identified factors associated with exercise participation and adherence in a sample of 102 sedentary, functionally limited, community-dwelling adults aged 60 to 94 years who participated in a home-based resistance training program. Stepwise regression analyses revealed that baseline physical factors (i.e., higher levels of mobility, weaker muscle strength, and fewer numbers of new medical conditions) were associated with higher rates of participation in the home program. Positive attitudes and a sense of control toward exercise, lower levels of confusion and depressive moods, and the development of fewer new medical problems during the program were related to higher levels of adherence to the program. Findings revealed that although physical health variables were the primary indicators of an older person's overall participation in the program, it was the psychological factors that were most important to adherence to this home based program. PMID- 9726130 TI - Living arrangement transitions among America's older adults. AB - This analysis describes the relationship between age and transitions from four living arrangements: living alone, living with spouse only, living with a child, and living with a spouse and child. Data from the National Survey of Families and Households, collected in 1987-88 and 1992-93, are used to calculate destination specific hazard rates by age and then construct multiple-decrement life tables. Living alone or with a spouse are the most stable living arrangements during the early stages of later life, whereas for the oldest-old, living with a child is the most stable living arrangement. The young-old tend to exit living arrangements through changes in coresidence, whereas transitions among the oldest old are primarily due to institutionalization and death. PMID- 9726131 TI - Historical and contextual correlates of parallel services for elders in African American communities. AB - Employing ethnographic and historical research methods, this article examines the organization and delivery of parallel services (locally generated alternatives to formal, externally controlled services) for older residents in four of the all Black towns of Oklahoma. These towns are rural, social-ecological enclaves that generally have remained totally African American since they were established in the late 1800s and early 1900s. Elders, even impaired older people, are prominent agents in the production of alternative services, and the organization of parallel services can be directly related to historical and contextual characteristics of the communities. The results demonstrate the value of taking into account community history and context, as well as the mix of parallel services, when developing formal services for minority communities. PMID- 9726132 TI - Special care units for demented patients: a multicenter study. AB - The local government of Regione Lombardia, Italy, recently (1994) funded a clinical and research project specifically devoted to dementia (Piano Alzheimer). A central role in this project has been reserved for the special care units (SCUs) for demented patients with behavioral disturbances. In order to evaluate their effectiveness, eight SCUs took part in this study. A specifically designed care program, focusing on environment and staff, was implemented in each SCU. Cognitive, functional, and somatic health status, and use of psychotropic drugs and of physical restraints were assessed at baseline, and after 3 and 6 months in 55 consecutively admitted patients. The data show an overall reduction in behavioral disturbances and a decreased use of psychotropic drugs and physical restraints. PMID- 9726133 TI - The role of religion/spirituality in coping with caregiving for disabled elders. AB - This study examined how religious/spiritual coping was related to specific conditions of caregiving and psychological distress among 127 informal caregivers to community-residing disabled elders. Support was found for the hypothesis that religious/spiritual coping influences caregiver distress indirectly through the quality of the relationship between caregiver and care recipient. Caregivers who used religious or spiritual beliefs to cope with caregiving had a better relationship with care recipients, which was associated with lower levels of depression and role submersion. PMID- 9726134 TI - Validation of the indicators of abuse (IOA) screen. AB - It is important to enable social service agency practitioners to identify cases in which seniors are abused by their caregivers. The Indicators of Abuse (IOA) screening measure provides an abuse screening tool, based on abuse indicators, for use by practitioners. The study (N = 341) supports the validity of the 29 item set of indicators of the IOA, which discriminates abuse cases (84.4% of the time) from nonabuse cases (99.2% of the time). An abuse-indicator model evolving from the IOA suggests three main types of abuse signals: (a) caregiver personal problems/issues; (b) caregiver interpersonal problems/ issues; (c) care receiver social support shortages and past abuse. PMID- 9726135 TI - Older adults and the news media: utilization, opinions, and preferred reference terms. AB - We surveyed 868 community-dwelling older adults about their (a) utilization of media news, (b) opinion of news media coverage of older people, and (c) preferences among terms used by news media to refer to senior citizens. Those with more than high school education read more newspaper news than those with less education. All groups watched TV news more frequently than they read the newspaper. Respondents had reservations about the news media's accuracy in, attitude toward, and interest in stories regarding seniors. Clear education and gender-related patterns emerged, whereas age proved nonsignificant in many analyses. Respondents' preferred reference terms were: (nouns) senior citizen, retiree, senior, and older adult; (adjectives) retired, senior, and mature. Respondents disliked: (nouns) old man/woman, old person, oldster, old timer, and geezer; (adjectives) old, aged, gray, and geriatric. PMID- 9726136 TI - Barriers to nutritional well-being for rural elders: community experts' perceptions. AB - Older adults use public and private services, as well as personal resources, to meet nutritional needs. In-depth interviews conducted with 73 service providers and community experts in two rural North Carolina counties were analyzed for these experts' perceptions of barriers to adequate nutrition for older adults. Perceived barriers included characteristics of the county and programs, transportation, and kin, as well as older adult medical and economic conditions, food habits, knowledge, and attitudes. The importance given each of these domains varied by respondents' area of expertise. Community experts and providers may not see the connection between their services and nutritional well-being of older adults. PMID- 9726137 TI - Building bridges between families and nursing home staff: the Partners in Caregiving Program. AB - Compelling evidence exists that conflict and communication problems occur between nursing home staff and family members of residents. However, few interventions have been documented that simultaneously address the needs of both groups. The Partners in Caregiving program was created to train staff and family members in communication techniques and conflict resolution skills. Through a joint meeting with facility administrators, both groups also have the opportunity to influence facility practices. Evaluation data indicated that satisfaction with the program was extremely high, and that positive changes in staff-family interactions occurred. PMID- 9726138 TI - Malaria: the call for action. PMID- 9726139 TI - Serum complement C4 levels during acute malarial infection and post-treatment period. AB - The researches are conducted in adults and children infected with the species P. falciparum and P. vivax. The results indicated that complement C4 level showed a marked fall during the course of acute malaria in both the children and adults infected with both species. A highly significant hypocomplementaemia was recorded (p < 0.01). Children infected with P. falciparum developed acute cerebral malaria, which was associated with marked hypocomplementaemia. A marked rise in C4 level was noticed on 10th day and it was found normal with reference to control, on 20th and 30th day. A considerable reduction in C4 complement was recorded in P. falciparum when compared to P. vivax. PMID- 9726140 TI - Intestinal parasites in the state of Bahrain. AB - Intestinal parasites infections are common in the state of Bahrain, but the incidence is decreasing due to improvement in health and social standards. A community-based study sample of all population of Bahrain was done including all ages and nationalities. From 1st July 1984 to 28th February 1986, the first community-based study sample of all population of Bahrain was done including all ages and nationalities. A total number of 2123, about 0.006 of the population of 1981 census was chosen for the study. The study shown that 739 persons were infected with intestinal protozoa and helminths. The figure representing 34.8% of total sample size, whom 283 persons were symptomatic, about 38.3% of total infected sample size. Giardia lamblia and Trichuris trichuria were the most common parasites among the infected persons. PMID- 9726141 TI - In vitro susceptibility of filamentous fungi to itraconazole. AB - The in vitro activity of itraconazole was investigated against 25 clinical isolates of filamentous fungi by agar dilution method. The isolated included Aspergillus sp., hyalohypomycetes, dematiaceous fungi and zygomycetes. Intraconazole was more active, inhibiting 50% (MIC 50) and 90% (MIC 90) of the Aspergillus sp., at a concentration of 0.5 and 2.5 ug ml-1 (MIC range 0.1 and 5 micrograms ml-1) Ketoconazole (MIC range 0.5-10 ug ml-1) required 1 and 5 ug ml-1 for inhibiting 50% and 90% of the isolates. For the hyalophypomycetes and dematiaceous fungi, the MIC 50s for itraconazole were 1 and 0.5 ug ml-1 and Ketoconazole required 2.5 ug ml-1 for both the groups. For the zygomycetes, the MIC range and MIC 50s for Ketoconazole were 1-100 and 10 ug ml-1 whereas the values for itraconazole were 5- > 100 and > 100 micrograms ml-1. PMID- 9726142 TI - A comparative study between rapid urease (modified), CLO test, culture and histopathological examination for Helicobacter pylori in patients with acid peptic diseases. AB - A modified Rapid urease test developed by us was evaluated as a screening test for Helicobacter pylori (H. pylori) during and endoscopy survey on patients with Acid Peptic Diseases (APD) and Non Ulcer Dyspepsia (NUD). This was compared with commercially available CLO (Campylobacter Like Organism) test, culture and histopathological examination. The modified Rapid urease test gave a sensitivity of 89.83% and a specificity of 100%, when compared to 95% sensitivity and specificity for commercially available CLO test. Our modified Rapid urease test is simple, economical and a quick test in identifying H. pylori in routine screening of patients with APD and NUD. PMID- 9726143 TI - Serotyping and transferability of drug resistance in clinical isolates of E. coli. AB - Forty nine multiple drug resistant strains of E. coli isolated from UTI were serotyped. The pattern was found to be 057 (eight strains); 0109 (four strains); 020, 038, 068, 0106, 0148. Rough (three each). 012, 054, 0101, 0160 (two each) and 02, 032, 046, 053, 060, 065, 090, 091, 0117, 0118, untypable (one each). The resistance pattern of all E. coli were identified and matted with recepient strain in penassay broth and in human urine. In a penassay broth transfer of resistance was demonstrated in 38 strains (77.5%) while in human urine transfer was demonstrated only in 14 strains (28.57%). PMID- 9726144 TI - Stool culture as a diagnostic aid in the detection of Entamoeba histolytica in the faecal specimens. AB - "The use of stool culture in Boeck & Drbohlav's biphasic amoebic medium as a routine diagnostic aid in the detection of Entamoeba histolytica in the faeces, is evaluated in the present study. A total of 3803 faecal specimens were examined for the presence of E.histolytica by direct smear, formalin ether concentration and culture during a study period of 1982-1990. A total of 259 stool specimens were positive for the parasite by any or all of these methods 42 (16.21%) stool specimen not cultured in Boeck & drbohlav's medium were possible by direct smear and concentration methods. 99 (38.22%) stool specimens were positive by all of these methods (direct smear, concentration and culture). The culture detected E. histolytica in additional 62 (23.93%) stool specimens which were negative by both the direct smear and concentration methods. Results of this study recommends the use of stool culture as a routine diagnostic aid in the laboratory, for the detection E. histolytica in the faeces". PMID- 9726145 TI - In vitro susceptibility of multiple drug resistant Salmonella typhimurium to newer fluroquinolone derivatives. AB - A total of 105 strains of Salmonella typhimurium resistant to Ampicillin, Co trimoxazole and Nalidixic acid were included in the present study. Among the new line of Fluroquinolones resistance to Ciprofloxacin (3.8%), Norflox (16.19%) and Ofloxacin (24.76%) was very low as compared to older antibiotics. All the 3 fluoroquinolones had MIC values less than 1 mcg/ml. PMID- 9726147 TI - Interobserver agreement in the diagnosis of cervical smears. AB - The present study was carried out to reveal the magnitude of individual variation in the diagnosis of Pap smears between two cytoscreeners and their compatability with cytopathologists and subsequent final diagnostic comparison with biopsy in 1,17411 cervical smears collected from different hospitals of Delhi during ten year period. Smears diagnosed as dysplasia at initial level by any one of the cytoscreeners were screened by cytopathologists for confirmation of diagnosis. An overall agreement of 94.9 percent was observed between two screens. 79.5 percent was agreement between screeners and cytopathologists. An agreement between cytology and histology in the diagnosis of dysplasia and malignancy were found to be 61.9 percent and dysplasia and malignancy were found to be 61.9 percent and 40.1 percent respectively. From the above study, it was observed that consistency among the two screeners was fairly acceptable. Keeping this observation in view, we can possibly practice two tier screening in place of three. PMID- 9726146 TI - Disk diffusion susceptibility testing of dermatophytes with imidazoles. AB - In vitro susceptibility testing of 43 isolates of dermatophytes was carried out against imidazoles-ketoconazole, miconazole and econazole and griseofulvin by agar dilution and disk diffusion methods. Econazole was the most effective drug inhibiting all the isolates at a concentration of 0.1 microgram ml-1. The MIC 50s and MIC 90s for ketoconazole and miconazole were 1 and 2.5 mg ml-1 whereas the values for griseofulvin were 1 and 5 micrograms ml-1. Good correlation was seen between the MIC and sizes of zones of inhibition around the disks. Regression analysis was used to measure the degree of correlation between the MIC values and matched averaged zones of inhibition and the correlation coefficients for econazole, ketoconazole, miconazole and griseofulvin were -0.5554, -0.5886, 0.8558 and -0.8268 (p < 0.001) respectively. PMID- 9726148 TI - Role of AgNORs in diagnosis of early malignant lesions of gall bladder. AB - Sections from eighty nine specimens of gall bladder including 40 cases of chronic cholecystitis, 9 dysplasia 15 well differentiated adenocarcinoma, 14 moderately differentiated adenocarcinoma and 11 poorly differentiated adenocarcinomas were studied for argyrophilic nucleolar organizer regions (AgNOR's). The difference between mean +/- SD NOR counts of chronic cholecystitis (1.97 +/- 0.28), dysplasia (5.6 +/- 0.88) well differentiated adenocarcinoma (7.2 +/- 0.30) and moderately differentiated adenocarcinoma (8.0 +/- 0.63) was statistically significant (p < 0.001). NOR counts of poorly differentiated adenocarcinoma (8.5 +/- 0.78) were higher than moderately differentiated carcinoma but the difference was not statistically significant. Despite the significant difference in the mean values, a considerable overlap in the NOR counts of individual cases of different groups was observed suggesting that though NOR counts can not act as a specific diagnostic parameter for diagnosis of early carcinoma and dysplasia in isolated cases, they may prove to be a good adjunct to already existing parameters like imaging techniques or cytology. PMID- 9726149 TI - Fine needle aspiration cytology of hepatocellular carcinoma. AB - Fine needle aspiration cytology (FNAC) is an extremely useful technique in the evaluation of hepatic masses. This study was undertaken with the aim of describing the morphological spectrum seen in fine needle aspirates from hepatocellular carcinoma (HCC) seen in our patients hailing from South India. Thirty two cases of HCC were studied. Trabacular pattern covered by endothelium was the most common. Pseudoglandular, spindle cell and dispersed patterns were also seen. Individual tumour cells were generally reminiscent of hepatocytes, and had a prominent nucleolus. The presence of intranuclear and intracytoplasmic inclusions were notable features. FNA cytology in HCC is sufficiently distinctive to form an invaluable tool in the diagnosis of this malignancy. PMID- 9726150 TI - Fine needle aspiration biopsy in monitoring human renal transplant. AB - Renal transplant dysfunction poses a diagnostic dilemma. Clinical evaluation frequently is inaccurate and needle biopsy carries a significant risk of bleeding. Fine needle aspiration biopsy (FNAB) being an easy to perform, a less traumatic, and rapid technique, was employed in 22 cases of clinically suspected renal allograft rejection. The results were described as nil rejection, mild and severe acute rejection, chronic rejection and cyclosporin nephrotoxicity. Seven of these cases were also needle biopsied. Histology confirmed the cytology findings in all. However, an additional finding of acute vascular episode was observed in a case of chronic rejection. Fine needle aspiration biopsy diagnosis helped in altering the immunosuppressive therapy in 16 graft rejections (excluding 2 cases of irreversible rejection) and 2 cases of cyclosporin toxicity. No rejection was found in 2 cases. Thus, 20 renal grafts could be brought back to normal function. Negligible incidence of complication viz. microscopic haematuria of short duration was noted in only 1 case. PMID- 9726151 TI - ACTH acts directly on melanocytes to stimulate melanogenesis--an in vitro study. AB - A total of 108 whole skin organ cultures taken from vitiliginous skin were incubated in MEM containing ACTH. It was observed that 53.7% that is 58 showed a positive response with an increase in pigment production and enzyme activity, as observed on frozen sections stained for dopaoxidase activity. On immunohistochemical staining for locating ACTH binding, it is observed that 27.3% control skin and 72.7% ACTH treated skin show positivity. The ACTH is seen to bind with the melanocyte membrane as well as the cytoplasm. This indicates that ACTH can bind to the MSH-receptors expressed by the melanocyte. Thus, ACTH acts directly on the melanocyte to enhance melanogenesis and does not require to act via the adrenal-pituitary axis. This also indicates that the response is not associated with immune suppression by ACTH. PMID- 9726152 TI - Effect of attachment surface on the growth and differentiation of breast cells. AB - This experimental work was undertaken to study the difference in cell growth due to different attachment surface. Three types of attachment surface were studied (i) collagenized surface (ii) glass surface and (iii) plastic surface. Rat tail collagen suspension was prepared and coated on culture flasks. Human breast epithelial cells and breast carcinoma cells were cultured for three weeks. Cell counts were made before and after one, two and three weeks of culture. Out of five cases only two survived for more than a week. The best survival was observed on plastic surface. Collagenized and glass surfaces had similar results. Thus plastic surface is probably the best compared to glass and collagenized surface, possibly because the plastic surface provides better adhesion to malignant cells. It was also seen that in plastic surface the cells get lined up around a basement membrane like structure growing a resemblance to ductal structure. This is contradictory to the prevalent view that collagenized surface is best for in vitro growth and differentiation. PMID- 9726153 TI - Phase contrast microscopic evaluation of placental pathology in premature gestation. AB - Numerous representative samples taken from forty placentas immediately after delivery from mothers associated with prematurity (i.e. less than 38 weeks), were semiquantitatively studied by phase contrast microscopy. Twenty placentas delivered from mothers without any antenatal complication served as control. Hypoplasia of the syncytium, stromal edema, ischaemia were prominent findings on phase contrast microscopy in the study group. Increased basement membrane thickening and high villous edema scores observed on light microscopy were statistically significant in prematurity as compared to controls. Observations by phase contrast and light microscopy were found complementary to each other. Phase contrast microscopy provided quicker results without disadvantage of fixation artefact and was found distinctly superior over the conventional histological methods. PMID- 9726154 TI - Granulomatous hepatitis: a retrospective study. AB - 51 cases of granulomatous hepatitis were seen among 1234 liver biopsies over a 10 year period. Tuberculosis was the commonest cause seen in 55 percent of cases. Other causes included leprosy, sarcoidosis, histoplasmosis, brucellosis, amoebic liver abscess, lymphoma and malignant granuloma. 12 percent of cases remained undiagnosed. Clinically these patients presented with pyrexia and hepatosplenomegaly. Jaundice was uncommon. Many showed elevated alkaline phosphatase levels, anaemia and raised ESR Granulomatous hepatitis of unknown aetiology with FUO was seen in 6 percent cases only. PMID- 9726155 TI - Cytodiagnosis of lower respiratory tract lesions by transthoracic needle aspiration. AB - A comprehensive prospective and retrospective study of 130 cases was carried out to evaluate the safety, adequacy and diagnostic accuracy of Transthoracic Needle Aspiration (TTNA) in various pulmonary lesions. Using both guided and unguided TTNA diagnostic material was procured in 86.15% cases. Rotex II, Chiba, lumber puncture and ordinary needles were used depending upon the type of lesion. Overall diagnostic accuracy of the procedure was 79.46 percent. Complication after the procedure were transient and self limiting. Haemoptysis was noted in 3.84 percent cases and pneumothorax in a single case. Thus TTNA can be safely included in the investigative protocol of lung lesions. PMID- 9726156 TI - Toxoplasma gondii IgM antibody prevalence study in patients suffering from neurological disorders. AB - One hundred and twenty seven patients belonging to Neurosurgery (49), Neuromedicine (48), Cardiac medicine (30) wards and Blood donors (30) as healthy controls were investigated for the prevalence of Toxoplasmosis by means of detecting specific IgM antibody against Toxoplasma gondii (T. gondii) employing Enzyme Immuno Assay (EIA). The detection rate of specific IgM antibody against T.gondii was found to be 32.7% (16/49) among Neurosurgery patients, 20.8% (10/48) among Neuromedical patients and 20% (6/30) among Cardiac medical patients. None of the voluntary blood donors tested was found to have T. gondii IgM antibody. Maximum prevalence rate was found among female patients undergoing Neurosurgery (42.3%) followed by Neuromedical patients (40%). There is an increasing rate of prevalence of Toxoplasmosis from the lower age group upto thirty years and a declining prevalence rate among the higher age groups. The present study revealed high prevalence rate of Toxoplasmosis in Neurosurgery patients (32.7%) and in particular among female (35.2%) than male (17.8%) patients. PMID- 9726157 TI - Epithelioid granulomas in Hodgkin's disease--prognostic significance. AB - Prognostic significance of non-caseating epithelioid granulomas in association with Hodgkin's disease has been studied. Such granulomas were found in 15 of the total of 104 cases of Hodgkin's disease encountered between Jan. 1981 and June 1990. These 15 patients were compared with 30 concurrent patients of Hodgkin's disease who did not have associated granulomas. All the patients were initially staged, treated and followed up for a period of two years. There was no significant difference in overall survival rate between the granuloma group and the control group. However, in relapse free survival rate in advanced stages of the disease (III & IV), although the difference between granuloma group and the control group was not statistically significant (p = 0.11), yet the relapse free survival curves revealed a tendency towards better survival with lesser number of relapses and longer remissions in granuloma group. PMID- 9726158 TI - Autosomal recessive polycystic kidney disease (ARPKD)--a report of two cases. AB - Autosomal Recessive Polycystic Kidney (ARPKD) is a very rare entity (1 in 15,000 live births) and mostly not compatible with life. Early diagnosis and genetic councelling may help prevent such births. Two interested cases are presented. PMID- 9726159 TI - Acute renal failure due to bilateral renal lymphoma. PMID- 9726160 TI - Diagnosis of malaria--an overview. PMID- 9726161 TI - High-performance membrane chromatography of supercoiled plasmid DNA. AB - Membrane adsorbers are well established in protein chromatography. The present paper investigated for the first time the behavior of polynucleotides on these stationary phases, taking a 7.2-kb predominantly supercoiled plasmid as example. Gradient and isocratic elution was studied. In contrast to protein high performance membrane chromatography (HPMC), isocratic elution is possible in DNA chromatography. In the case of gradient elution, much higher salt concentrations can be used in the starting buffer. Under optimized conditions, both approaches led to a splitting of the single plasmid peak into three maximums, which corresponded to the three-albeit isolated-bands in the agarose gel. Presumably the three fractions were supercoiled, nicked, and open circular plasmid DNA. Linearization of the plasmid lowered the adsorption energy, and the linearized plasmid eluted earlier than the nonlinearized one. The HPMC experiments were compared to similar ones performed using a conventional packed-bed anion-exchange column (BioScale Q2, 7 x 52 mm, 10-micron porous particles) and a novel monolithic-type anion-exchange column (UNO Q1, 7 x 35 mm). The results and characteristic differences observed in these experiments were interpreted in the light of the newly developed theory of HPMC. PMID- 9726162 TI - In situ antigen immobilization for stable organic-phase immunoelectrodes. AB - A new method based on enzymatic single-step in situ synthesis of hapten-carrier conjugates on electrodes is described yielding stable, reproducible, and reusable organic-phase immunoelectrodes (OPIEs). The electrodes developed were tailored for analyte detection in organic solvents and allow for the analysis of soil extracts without further sample processing and cleanup. Catalyzed by transglutaminase from a variant of Streptoverticillium mobaraense, the reaction proceeds in aqueous solution with and without addition of organic media in only 1.5 hours. In this study, the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) was chosen as model compound and chemically amino-functionalized prior to its enzymatic immobilization. The high reproducibility of the immobilization procedure allowed for batch calibration of the immunoelectrodes. Moreover, pure methanol or treatment with diluted sulfuric acid used for regeneration studies did not disturb the hapten layer. The OPIE consists of screen-printed carbon electrodes, monoclonal anti-2,4-D antibodies, and the immunochemical recognition reaction and was optimized with regard to a high stability in organic media. For electrochemical detection, horseradish peroxidase was used as enzyme label together with H2O2 as substrate and hexacyanoferrate (II)/(III) as mediator. The OPIE showed high stability upon storage over 93 days. Response times of 17 s (t95) were found to be advantageous compared to those of other biosensors. Including the immunochemical reactions, the complete assay takes 30 min. A calibration curve for 2,4-D in 30% methanol/buffer obtained with 70 electrodes within 4 weeks revealed a detection limit of 9 mg/L, a sensitivity of 1.3 nA L mg 1 cm-2, and a repeatability of 6.8%. Although we calculated a lowered repeatability for reused electrodes of 13.4% and a slightly decreased sensitivity of 0.9 nA L mg-1 cm-2, multiple-used OPIEs could also be applied for calibration. PMID- 9726163 TI - Cyclic voltammetric simulation for electrochemically mediated enzyme reaction and determination of enzyme kinetic constants. AB - A cyclic voltammetric simulation that can be applied to an electrochemically mediated enzyme reaction involving any substrate and mediator concentrations was developed. Concentration polarization of the substrate in the vicinity of an electrode was considered as well as mediator concentration. Reversible and quasi reversible electrochemical reactions with one electron followed by an enzyme reaction with two electrons were modeled. The differential equations for the mediator and substrate were solved using digital simulation techniques. The calculated cyclic voltammograms showed prepeaks when there was a low substrate concentration, high mediator concentration, and high enzyme activity. Digital simulation was applied to the determination of the kinetic constants of glucose oxidase (GOx). Cyclic voltammetry was carried out experimentally in a phosphate buffer solution containing GOx, ferrocene derivatives, and glucose. The ratio of the catalytic to the diffusion-controlled current, ik/id, was evaluated. The kcat, KMM, and KMS values were determined from the current values obtained by simulation and by experimentation at various enzyme, mediator, and substrate concentrations. The kcat, KMM, and KMS values for GOx, ferrocenedimethanol, and glucose were 340 s-1, 110 microM, and 30 mM, respectively. PMID- 9726165 TI - Optical time-of-flight chemical detection: absorption-modulated fluorescence for spatially resolved analyte mapping in a bidirectional distributed fiber-optic sensor. AB - A continuous chemically sensitive optical fiber is used with optical time-of flight chemical detection (OTOF-CD) for spatially resolved analyte mapping. To enhance signal levels and to improve their reproducibility, two novel principles for signal generation and processing are introduced. In the first, the fluorescene of an analyte-insensitive fluorophore is monitored as a function of the evanescent wave absorption of an analyte-sensitive indicator. The resulting signal levels are well above those encountered in optical time domain reflectometry methods that rely upon backscattering for spatially resolved detection. As a result, the method could significantly expand the range of species that can be detected with absorption reagents used in OTOF sensors. The second method raises signal-to-noise ratios by 3-4.5-fold for measurements made at the far ends of the sensing fiber. It functions by sending probe laser pulses into and monitoring their return sequentially from both ends of the sensing fiber. Because the two pulses provide complementary information, only the first half of each of the collected waveforms is used for analyte quantitation. The introduced concepts were experimentally verified with a distributed sensor constructed from a 40-m-long continuous chemically sensitive optical fiber. This sensing element was produced by immobilization of an ammonia-sensitive absorbing reagent (phenol red) and an analyte-insensitive fluorophore (rhodamine 640) into the original silicone cladding of the plastic-clad silica fiber. PMID- 9726164 TI - A parallel multiharmonic frequency-domain fluorometer for measuring excited-state decay kinetics following one-, two-, or three-photon excitation. AB - We report on the performance of a new, multiharmonic frequency-domain instrument that uses the high harmonic content of a passively mode-locked, pulse-picked femto-second Ti-sapphire laser as the excitation source for the determination of one-, two-, or three-photon excited time-resolved fluorescence anisotropy and intensity decay kinetics. In operation, the new instrument can provide a complete frequency-domain data set at 100 modulation frequencies in less than 1 min. The new instrument exhibits 5-10-ps measurement precision and it can rapidly and accurately recover complex excited-state fluorescence anisotropy and intensity decay kinetics under one-, two-, or three-photon excitation for dilute or optically dense samples that exhibit single or multiexponential decay kinetics. This latter aspect of the instrument is demonstrated by successfully determining the excited-state intensity decay kinetics for a dilute aqueous solution of rhodamine 6G dissolved in a high concentration of bromocresol green. This approach is extended by determining the excited-state fluorescence intensity decay kinetics of dilute fluorescein directly in undiluted, whole blood as a function of pH under two-photon excitation conditions. The high-speed capabilities of the new instrument are exploited by performing two-photon excited fluorescence anisotropy decay experiments on the fly for site-selectively labeled bovine serum albumin as it undergoes enzymatic digestion by trypsin. PMID- 9726166 TI - On-the-fly fluorescence lifetime detection of dye-labeled DNA primers for multiplex analysis. AB - Mixtures of dye-labeled, M13-forward DNA primers were separated by capillary gel electrophoresis and detected on-the-fly, using fluorescence lifetime measurements, to evaluate four-decay detection for multiplex DNA sequencing. Three different four-dye systems were used, two that were excited at 488 nm and one that was excited at 514 nm. Each dye-labeled primer was identified on the basis of the lifetime of the conjugated dye using nonlinear least squares or the maximum entropy method to analyze the lifetime data. Overlapping electrophoretic peaks were generated by making multiple injections of mixtures of the dye-labeled primers. The overlapping peaks were resolved by fitting the data to two-, three- or four-component lifetime models used in nonlinear least-squares analysis in which each lifetime component was fixed to the predetermined lifetime of the corresponding dye-labeled primer. In two of the dye systems, the lifetimes of the four dye-labeled primers were sufficiently different to allow peak resolution. In the other dye system, addition of 10% DMSO to the run buffer changed the lifetime of one dye-labeled primer, allowing it to be resolved from another dye-labeled primer with similar lifetime. PMID- 9726167 TI - Adapting selected nucleic acid ligands (aptamers) to biosensors. AB - A flexible biosensor has been developed that utilizes immobilized nucleic acid aptamers to specifically detect free nonlabeled non-nucleic acid targets such as proteins. In a model system, an anti-thrombin DNA aptamer was fluorescently labeled and covalently attached to a glass support. Thrombin in solution was selectively detected by following changes in the evanescent-wave-induced fluorescence anisotropy of the immobilized aptamer. The new biosensor can detect as little as 0.7 amol of thrombin in a 140-pL interrogated volume, has a dynamic range of 3 orders of magnitude, has an inter-sensing-element measurement precision of better than 4% RSD over the range 0-200 nM, and requires less than 10 min for sample analysis. The aptamer-sensor format is generalizable and should allow sensitive, selective, and fast determination of a wide range of analytes. PMID- 9726168 TI - Cerenkov radiation as a UV and visible light source for time-resolved fluorescence. AB - We demonstrate the first use of Cerenkov radiation for the measurement of fluorescence lifetimes. Relativistic beta particles from the nuclear decay of 90 Sr and 90Y generate a spectral continuum in a quartz waveguide. Light flashes of < 100-ps duration are delivered simultaneously to a sample cell and a reference photomultiplier. A simple, digitally based cross-correlation signal processor allows extraction of the sample fluorescence decay kinetics without distortions which can result from the random excitation pulse sequence. We characterize both the pulse duration and the pulse intensity of the light that is emitted from the waveguide. Although the excitation intensity is very weak, we demonstrate that accurate lifetime measurements are possible with only a few hundred seconds of integration time. Tests on a variety of compounds illustrate the utility of the light source throughout the UV and blue regions of the spectrum. We also discuss future design improvements and potential applications of this new approach to time-resolved fluorescence. PMID- 9726169 TI - Analytical performance of accelerator mass spectrometry and liquid scintillation counting for detection of 14C-labeled atrazine metabolites in human urine. AB - Accelerator mass spectrometry (AMS) has been applied to the detection of 14C labeled urinary metabolites of the triazine herbicide, atrazine, and the analytical performance of AMS has been directly compared to that of liquid scintillation counting (LSC). Ten human subjects were given a dermal dose of 14C labeled atrazine over 24 h, and urine from the subjects was collected over a 7 day period. Concentrations of 14C in the samples have been determined by AMS and LSC and range from 1.8 fmol/mL to 4.3 pmol/mL. Data from these two methods have a correlation coefficient of 0.998 for a linear plot of the entire sample set. Accelerator mass spectrometry provides superior concentration (2.2 vs 27 fmol/mL) and mass (5.5 vs 54,000 amol) detection limits relative to those of LSC for these samples. The precision of the data provided by AMS for low-level samples is 1.7%, and the day-to-day reproducibility of the AMS measurements is 3.9%. Factors limiting AMS detection limits for these samples and ways in which these can be improved are examined. PMID- 9726170 TI - Quantitative determination of biological sulfhydryl groups by postcolumn derivatization and elucidation of microheterogeneity of serum albumins. AB - A quantitative analytical system for biological sulfhydryl compounds has been developed using an ion-pair reagent with isocratic elution and an on-line postcolumn derivatization with Ellman-type reagents. As human or bovine serum albumin has 35 cysteinyl residues, one cysteinyl residue exists as a free sulfhydryl moiety, and this gives rise to the microheterogeneity in serum albumin. Here we report for the first time the quantitative characterization of the microheterogeneity of serum albumin. Cysteine was found to be the major molecule attached to the sulfydryl group of the serum albumins. Although glutathione could not be detected, the Cys-Gly element of glutathione was found. Freshly prepared human serum albumin from healthy volunteers contained 0.46 nmol of Cys/mL of serum, 0.24 nmol of Cys-Gly/mL of serum, and very small amounts of glutathione (0.02 nmol/mL). PMID- 9726171 TI - Competitive binding assay for thyroxine using in vitro selected oligonucleotides. AB - A new binding assay that uses oligodeoxyribonucleotides (DNAs) obtained by the in vitro selection method, instead of antibodies, to bind to the target molecule, thyroxine (T4), is described. The DNAs which selectively bound to the T4 were selected, labeled with biotin or radioisotope, and then used to detection of T4 in the presence of liothyronine (T3), which has a chemical structure similar to that of T4. PMID- 9726172 TI - Mass spectrometry. PMID- 9726173 TI - Nursing leadership and autonomous professional practice of registered nurses. AB - Autonomous professional practice continues to be elusive for registered nurses. Autonomous professional practice implies that nurses would be free to determine the procedures for carrying out their nursing work. In other works, they would be able to make independent decisions about their own nursing practice. This article reports research that describes the nature of nursing leadership that supports autonomous professional practice of registered nurses. PMID- 9726174 TI - Perspective: the Internet--an invaluable career planning and development resource. PMID- 9726175 TI - The emergence of a culture that promotes evidence based clinical decision making within an acute care setting. AB - Nursing research programs within acute care hospitals are essential to the development and integration of nursing knowledge, difficult to implement and rarely evaluated. The purpose of this paper is three fold: (1) to describe the development, structures, and processes of a nursing research program within an acute care teaching hospital and (2) to describe selected evaluation outcomes and (3) to discuss future directions. PMID- 9726176 TI - Research utilization: evaluation of initiatives in a public health nursing division. AB - The need to incorporate research findings into public health nursing has never been greater. This paper describes briefly, the initiatives to promote research in a public health nursing division and the results of the evaluation questionnaire. Results indicated that public health nurses valued research and felt comfortable with the concepts and phases of the research utilization model. They would engage in research activities if conducted at team meetings and when time was allotted. They identified administrative support and the supportive environment as being positive facilitators to research utilization. Despite these findings, the majority (67.5%) were not changing their practice as a result of the initiatives. They identified having difficulty formulating a research question and needing assistance with article critique. Time was cited as the greatest deterrent. They felt they did not have time to read research or engage in the steps of the research utilization model. Recommendations from the evaluation include the need to designate time for research utilization at team meetings. Once the nurses comfort level and value placed on research utilization increases, they may be motivated to initiate research activities on their own. PMID- 9726177 TI - Home care quality management. Where are we now? Where do we go from here? AB - This article is the first of a series of three focusing on key management processes in home care--quality management, financial and information management, and, service and human resource management. The articles examine the current state of practice in Ontario and suggest a desirable future state. In this first article, home care is defined and a brief description is provided from a national and provincial perspective. Quality is also defined and the drivers for quality in home care outlined. Using a framework of key principles, the current home care program is assessed and conclusions drawn about where it sits on the quality continuum. Activities to move home care closer to true quality management are then suggested for the province, community, and service provider levels. PMID- 9726178 TI - Case management: a literature review. AB - The achievement of an integrated system of care is one of the major goals of Canadian health care restructuring. Although many of the structural barriers to service integration have been removed, integration of care for individuals with long-term, complex health and human needs has not yet been achieved. Case management is now being considered in many countries as a method of integrating and coordinating health and social service systems. Central to the effectiveness of case management is the role of the case manager. The case manager provides clients with continuity, consistency, and coordination of care across all clinical settings and boundaries. Comprehensive case management practice requires professionals with the knowledge and skills to work within existing bureaucratic and organizational service systems. This article is a summary of the literature on long-term care case management, with a focus on Canadian studies. PMID- 9726179 TI - From nursing data to information to evidence: are we prepared? PMID- 9726180 TI - Will evidence-based nursing practice make practice perfect? AB - Evidence-based practice, or evidence-based decision-making, is rapidly developing as a growth industry in nursing and the health professions more widely. It has its origins in the work of the British epidemiologist Archie Cochrane and has recently been re-energized in Canada by the National Forum on Health and its call for a culture of evidence-based decision-making. Before we adopt evidence-based nursing (EBN) as a mantra for the 21st century, we should examine its origins and its consequences, and we should probe related concepts, 2 of which are the nature and structure of practice-based knowledge and the nature and structure of evidence generally. Findings of a recent survey of nurses in western Canada are used to illustrate that nurses use a broad range of practice knowledge, much of which is experientially based rather than research-based. PMID- 9726181 TI - Implementation of a clinical information system in nurse-managed care. AB - The Penn Nursing Network Information System Project is a collaborative effort of practitioners, academic researchers, and a health-care software developer. The Penn Nursing Network, a group of nurse-managed practices owned and operated by the University of Pennsylvania School of Nursing, has taken a leadership role in the project. PNN is developing an information system specific to the needs of nurse-managed care and creating a data warehouse for nursing centres in the Philadelphia region. Important components of this project include the identification of key data elements to represent the problems treated, interventions performed, and outcomes sensitive to the nursing care provided. The Omaha System provided a useful framework for capturing the necessary data elements. However, additional data were needed. In addition, attention was paid to the development of a software program that would complement the workflow of the practitioner while capturing data efficiently. The main goal of the project is development of a longitudinal database reflective of clinical practice, to be used for both research and evaluation. PMID- 9726183 TI - Nursing informatics. PMID- 9726182 TI - Classification systems for health concerns, nursing strategies, and client outcomes: nursing practice with families who have a child with a chronic illness. AB - This paper describes 3 classification systems developed from a study of the effectiveness of a nursing intervention in improving the psychosocial adjustment of children with a chronic illness. The study nurses' documentation of the nursing care provided to the 163 participating families was content analyzed. Systems were developed to classify the types of: (a) health concerns or issues that were the focus of the nursing, (b) actions the nurses used to help families achieve their goals, and (c) outcomes observed by the nurses. These classification systems have furthered our understanding of the McGill Model of Nursing, and they describe the scope of nursing practice based on this nursing perspective with a particular population (families who have a child with a chronic illness). These systems could be used to describe and measure nursing practice with this and other groups of clients. PMID- 9726184 TI - Stressors in families with a child with a chronic condition: an analysis of qualitative studies and a framework. AB - This article presents a comprehensive, research-based, clinically useable framework of stressors and tasks for families with a child with a chronic condition. Terms such as stressors, tasks, challenges, concerns, and problems are commonly used to describe the struggles and triumphs of these families. Their unique and changing nature has complicated comprehensive description. The steps in the early development of the Burke Stressors and Tasks Framework for Families with a Child with a Chronic Condition, and its clinical uses, are briefly described. The final step in the development of the Framework is discussed in more detail. This was a "meta-analysis" of qualitative research findings that confirmed the components of the Framework. Conclusions are drawn for subsequent research steps and clinical uses of the Framework. PMID- 9726185 TI - Falls risk factors in an acute-care setting: a retrospective study. AB - Research findings have been contradictory regarding risk factors for falls in the acute-care setting. Identification of factors that place individuals at risk of falling in this setting are a priority because falls result in high morbidity and mortality and thus increased healthcare costs. The purpose of this study was to extend knowledge beyond the known risk factors of age and medical diagnosis by comparing the characteristics of 301 adults who fell while hospitalized with a matched sample of adults who did not fall while hospitalized. A descriptive, retrospective, comparative design was used. The fall and non-fall group were matched on age and primary medical diagnosis at the time of discharge. Data were collected from hospital incident reports and medical records. Logistic regression for matched groups identified 5 risk factors, as follows. Incontinence. The odds of falling were 11.3 (CI = 3.85, 33.05) times greater for those who were incontinent than for those who were not incontinent. Long hospital stay. The odds of falling were 9.9 (CI = 4.89, 19.88) times greater for those hospitalized 19 days or longer than for those hospitalized less than 19 days. Dependency for ambulation. The odds of falling were 6 (CI = 2.83, 12.84) times greater for those who were dependent for ambulation than for those who were independent. Independency for hygiene. The odds of falling were 2.5 (CI = 1.23, 4.88) times greater for those who were independent for hygiene than for those who were dependent. Lack of regular exercise. The odds of falling were twice as high (CI = 1.00, 3.82) for those who did not exercise regularly as for those who exercised regularly. These findings suggest that ongoing assessment may be more important than the admission assessment in identifying risk factors for falls in the acute care setting. No 2 studies have found exactly the same set of risk factors, although some findings are consistent across studies. This suggests that those risk factors that are consistent across studies may identify persons who are at the greatest risk for falls and that other risk factors for falls are specific to a patient population. PMID- 9726186 TI - [Quality of life perception by women suffering from stage III or IV primary pulmonary hypertension and receiving prostacyclin treatment]. AB - The goal of this study was to examine perception of quality of life in women suffering from stage III or IV primary pulmonary hypertension and being treated with prostacycline. This medication is a non-specific vasodilator with a short half-life. This was an explorative and descriptive multiple-case study (3 cases). The interview guides were elaborated according to Zhan's conceptual model of quality of life (1992). These helped in collecting qualitative data for content analysis. The results indicated that the 3 women were not satisfied with their quality of life, in terms of physical, psychological, social, and economic well being, during the 12 weeks following the initiation of their treatment. Even though medical studies have shown a decrease in the mortality rate of individuals suffering from primary pulmonary hypertension within the first 12 weeks of prostacycline treatment, this case study cannot conclude that the quality of life of these 3 women had improved. A longer data collection period, with a larger sample size of patients, is recommended for future research. PMID- 9726187 TI - From rhetoric to action: establishing community participation in AIDS-related research. AB - Acquired Immunodeficiency Syndrome (AIDS) has been described as the most challenging disease in modern history. For many people with HIV/AIDS (PWA), issues of appropriate respite care and supported housing are a pressing concern. To meet the housing requirements for PWAs, it is essential to engage this community in determining its own housing needs. To that end, a participatory action research (PAR) investigation was undertaken with the PWA community to ascertain the desirability and feasibility of a supported-living/respite-care community home. PAR has been heralded as an important research methodology in ensuring research relevance and community participation leading to effective change. Although nursing interest in PAR is increasing, there are few nursing examples that describe the process of undertaking PAR. The purpose of this article is to describe, explain, and critique the use of PAR in the case of a supported-living/respite-care home. A PAR framework is presented and the process of conducting PAR is outlined. PMID- 9726188 TI - Courage in middle-aged adults with long-term health concerns. AB - The purpose of this study was to develop a substantive grounded theory of courage among middle-aged adults with long-term health concerns. Twenty-five persons from rural and non-metropolitan areas of Central Illinois were selected to participate in this study based on theoretical sampling procedures. Interviews of 1 to 2 hours using openended questions were audiotaped and transcribed verbatim. The data were analysed using grounded theory methods. Courage among middle-aged adults with long-term health concerns was determined to consist of an ongoing progressive-regressive process of becoming and being courageous. Being courageous involves being fully aware of and accepting the threat of a long-term health concern, solving problems using discernment, and developing enhanced sensitivities to personal needs and the world in general. Courageous behaviour consists of taking responsibility and being productive. Courage is not limitless, and the process of becoming and being courageous is dependent on intrapersonal and interpersonal factors. Health-care providers facilitate this process by demonstrating competence and communicating effectively. Outcomes of being courageous include personal integrity and thriving in the midst of normality. PMID- 9726189 TI - Canadian nurses' views on assignment of publication credit for scholarly and scientific work. AB - A total of 184 Canadian nurses who were expected to publish scholarly and/or scientific work or whose roles provide for socialization of nurses in academic endeavours, research, and publication were asked to respond to 42 scenarios. This study replicated, with some modifications, surveys conducted in 1981, 1985, and 1987 to determine the views of American nurses on assignment of publication credit. The scenarios in the present survey required judgements about how authorship and footnote credit should be allocated among groups involved in research and academic writing; in some scenarios all the individuals were nurses (in both clinical and academic settings), while other scenarios featured collaboration between nurses and other health-care professionals or focused on interactions between nursing professors and students. While consensus of greater than 80% was achieved for only 7 of the 42 scenarios (modal responses), the respondents' written comments revealed 2 recurrent themes: that credit should be based entirely on contribution, rather than status; and that, as much as possible, authorship and footnote acknowledgement should be discussed and resolved before contentious issues arise. There was widespread agreement on these 2 principles. However, there was disagreement concerning collaborative academic work, particularly concerning the forms of collaboration that merit authorship credit and the forms that are sufficiently acknowledged through footnoting. Both the model responses and the areas of disagreement are discussed. PMID- 9726190 TI - Sudden unexplained death in epilepsy: what people with epilepsy and their families need to know. PMID- 9726191 TI - Ethical dilemmas in people with epilepsy who drive. AB - Concern about driving safety in people with epilepsy has been present since the automobile became a widely used source of transportation (Siegel, 1988). Unfortunately, we still do not have adequate means to determine who can and cannot drive. Clinicians must use their best medical judgment to make these determinations. They then must consider the aspects of their state's driving laws and legal consequences of their judgments. In the 44 states that do not require clinician reporting of seizures, they must deliberate about breaching or maintaining the confidentiality of patients with uncontrolled seizures who continue to drive. In making this deliberation they should consider both the probability and magnitude of harm posed by continued driving of an individual patient. Judgments to breach confidentiality should include assessment of risks and benefits to the patient and to society by taking such action. Ultimately, each patient must be judged individually. PMID- 9726192 TI - Preventing accidental injury in people with epilepsy. PMID- 9726193 TI - Living safely with seizures: assistive technology, rehabilitation technology, and resources. PMID- 9726195 TI - The evolution of the auditory midlatency response in evaluating unconscious memory formation during general anesthesia. AB - The inability to objectively evaluate the amnesic status of an anesthetized patient has been a perplexing problem for the anesthesia provider. One approach thought to be effective in evaluating the amnesic status of the anesthetized patient is the auditory midlatency response (AMLR). The AMLR is an electrophysiological response that is recorded from scalp electrodes 10 to 80 ms after the auditory pathways begin to process acoustic stimuli. The response is thought to reflect the anesthetized patient's ability to consolidate an acoustic stimuli into an explicit or implicit memory. This article defines the amnesic state, describes the various components of AMLR, discusses clinical uses of the AMLR as an indicator of memory formation during the anesthetized state, and explains the clinical implications of using the AMLR in the surgical suite. Recent results have noted that the Pa waveform, the first positive deflection of the AMLR, may be the component that may serve as an intraoperative indicator of the anesthetized patient's ability to potentially consolidate an intraoperative acoustic stimuli into a memory. With the establishment of the Pa waveform of the AMLR as a reliable indicator of intraoperative memory formation, the AMLR can then be used to significantly decrease the occurrences of traumatic neurosis in the surgical patient and subsequent medicolegal consequences for the health care team. Thus, the use of the AMLR strives to promote a safer intraoperative environment for both the patient and the anesthesia provider. PMID- 9726194 TI - Awareness during general anesthesia: new technology for an old problem. AB - The possibility of awareness during general anesthesia causes apprehension for the patient and the Certified Registered Nurse Anesthetist (CRNA). The goals of general anesthesia are to prevent the sensation of pain and produce a state of sedation, hypnosis, and unconsciousness so the patient will not remember the surgical procedure. An inadequate level of anesthesia can result in patient awareness during surgery. The current practice of anesthesia relies on indirect hemodynamic measurements such as blood pressure and heart rate to monitor the sedative hypnotic state of the patient's brain during general anesthesia. Hemodynamic responses are not reliable for predicting awareness just as blood pressure and heart rate are not indicative of consciousness. Electroencephalogram (EEG) waveforms are known to be affected by anesthetics. Characteristic EEG waveforms are a direct indication of the patient's level of consciousness. Unprocessed and computer-processed EEG recordings have been used in an attempt to monitor the patient's level of consciousness during general anesthesia. A raw or unprocessed EEG recording to monitor the level of consciousness during general anesthesia is problematic. The EEG signal is complex, affected by artifact, and it requires a dedicated interpreter. Conventional processed EEG monitoring systems are problematic because of the complexity of the equipment and technical difficulty of reading the EEG recording. The purpose of this article is to describe the history of awareness during anesthesia and introduce a new processed EEG monitor, the Bispectral Index (BIS) (Aspect Medical Systems, Inc., Natick, MA) with implications for future clinical use and research. PMID- 9726196 TI - Planning and implementing an anesthesia crisis resource management course for student nurse anesthetists. AB - Development of Anesthesia Crisis Resource Management (ACRM) skills is a highly desirable outcome of a nurse anesthesia educational program. Access to an ACRM course is limited by a variety of factors including cost, availability of a center, time constraints, and the lack of adequately prepared CRNA faculty. The authors describe a Nurse Anesthesia Program's planning and implementation of an ACRM course by using a high fidelity human simulator. Key supportive and logistical elements necessary to implement an ACRM course are described. The authors address administrative concerns, faculty preparation needs, and explain the process of course design. Strengths of the course and problems encountered during the implementation of the experience are described. Feedback from participating students, faculty perceptions, and lessons learned from the experience are shared. PMID- 9726197 TI - The use of an anesthesia simulator in graduate and undergraduate education. AB - The use of simulation for educational training is common in many industries but is a new advancement in student instruction in anesthesia, medicine, or allied health. The Human Patient Simulator (HPS) allows students and clinicians to learn and practice a variety of technical skills as well as manage basic and complex clinical situations in a modifiable and reproducible environment. The HPS has been extensively integrated in many graduate and undergraduate Nursing and Allied Health courses at the Medical College of Georgia. Use of the HPS stems from the theory of situated cognition, which states that students best learn "what to do" and "how to do" in a real world environment. The HPS provides a real world environment for student learning in various fields. The theory of situated cognition is discussed in conjunction with the implementation of the HPS into various classes. PMID- 9726198 TI - Anesthesia for an achondroplastic dwarf with bilateral vocal cord granuloma: use of a Xomed Hunsaker Mon-Jet ventilation tube. AB - Technological advances have greatly improved the management of the patient undergoing microlaryngeal surgery. The use of a laser, high frequency jet ventilation (HFJV), total intravenous anesthetic techniques (TIVA), and specially designed endotracheal tubes (Xomed Hunsaker Mon-Jet ventilation tube [Xomed Surgical Products, Jacksonville, FL]) are recognized as cutting edge approaches to the management of these cases. The use of high technology adjuncts requires considerable skill and knowledge from the perspective of the anesthesia provider. This case report describes an approach to anesthetic management for a patient with a history of achondroplastic dwarfism having laser excision of bilateral vocal cord granulomas. Further increasing the complexity of the case was the surgical use of an autogenous tissue glue, which required a period of 'airway silence' during the application process. PMID- 9726199 TI - Quarterly update. Anesthetic drug interactions. PMID- 9726201 TI - JCAHO revised interpretation PMID- 9726200 TI - Where do your legislative representatives stand on the issue of home care? PMID- 9726203 TI - Mail-order pharmacy. Teaching your patients to use it safely. AB - People of all ages are discovering the convenience and savings of mail-order pharmacy use. On the surface, this option seems like a commonsense way to save money on healthcare, but the use of mail-order pharmacy among the elderly may increase the risk of unintentional medication noncompliance. This article discusses the advantages and risks of using this service and presents guidelines that can maximize the safety of mail-order pharmacy. PMID- 9726202 TI - The impact of the Balanced Budget Act of 1997 on home healthcare agencies and nurses. AB - "Medicare home health agencies are facing a regulatory purgatory as measures to reduce home health spending by about $16 billion over five years take effect with little government guidance. Home health providers are grappling with new surety bond requirements, the banning of venipuncture as a qualifying procedure for other home health services, and an interim payment system to scale back reimbursement levels before a prospective payment system takes effect in fiscal year 2000" (Cassil, 1998). The challenges of surviving and conquering all of the current issues facing home healthcare nurses and agencies seem overwhelming. All of these legislative and regulatory issues with the resulting financial and clinical implications are occurring as agency staff and administrators continue to provide quality home healthcare services to patients of all ages. Please share your strategies for meeting the challenges of 1998 with the editor. PMID- 9726204 TI - The 4 rights of compression therapy for patients with chronic venous insufficiency and venous ulceration. AB - Compression therapy has become an ever increasing component in the care of patients with Chronic Venous Insufficiency (CVI) and venous ulcerations. Although compression therapy is of great benefit to many of these patients, this treatment option does not come without risks. The increased use of compression therapy has coincided with an increase in its improper application. This article simplifies the principles of compression and highlights the common pitfalls of its use, with an emphasis on the home healthcare setting. PMID- 9726205 TI - Cardiovascular assessment for home healthcare nurses. Part 1: Initial cardiovascular assessment. AB - A description of the technique for standard cardiovascular assessment as it applies to patients in the home care setting is provided in this two-part article. The first, Initial Cardiovascular Assessment, covers basic physical assessment of the cardiovascular patient. The second article, Assessing Blood Pressure and Cardiac Function, which will appear in the August, 1998 issue, provides information that home care nurses can use to develop a thorough method of blood pressure, heart, and breath sound assessment. PMID- 9726206 TI - Does your home healthcare client have lymphadenopathy? AB - The routine head-to-toe home care assessment should include screening for abnormal lymph nodes. This can provide critical information about existing disease states or underlying problems that may have been overlooked. This article details the lymph node examination for the home care client that is easily incorporated into the initial examination, allows for fast documentation of findings, and provides helpful assessment information for disease detection in adults and children. PMID- 9726208 TI - The ABCs of a job interview. PMID- 9726207 TI - Lives on hold: evaluation of a caregiver's support program. AB - Evaluation research of a caregiver community-based support program is described. Initially, caregiver-participants characterized their lives as being "on hold." They were overwhelmed, isolated, and lacking time for a personal life. Socialization activities and learning to care for themselves were the most powerful and helpful aspects of the program. Suggestions are made for developing a caregivers' support program within a home healthcare agency. PMID- 9726209 TI - Keeping Steve at home. Coordinating services for long-term benefits. PMID- 9726210 TI - Florence Nightingale's lamp ... 1998. PMID- 9726211 TI - Change: managing transition. PMID- 9726212 TI - A postoperative anoxic event that ended life. PMID- 9726214 TI - Acute bereavement services and routine referral as a mechanism to increase donation. AB - Efforts to develop a mechanism that improves donation rates and provides better service to patients and staff are presented in this article. The University of Nebraska Medical Center has incorporated a routine referral process into its acute bereavement services requiring representatives to respond to each in-house death to provide consistent support and management for the decedent's family and the hospital staff. Every family was offered bereavement support and the opportunity to consent to autopsy as well as to organ and tissue donation if medically appropriate. Key data related to death and consent discussions were documented, routinely reviewed, and reported to a central location. Appropriate and timely access of these data helped to modify the program and assess its need for additional education or intervention. PMID- 9726213 TI - Analysis of donor versus nondonor demographics. AB - A retrospective analysis of the demographic features of all potential organ donors over a 3-year period (1994-1996) at one organ procurement organization was conducted. The potential donor pool of 495 people was 42% female and 58% male, with a slight difference in consent by gender. The mean income difference between donors and nondonors was less than $3000 per year (obtained from zip code census data). Educational achievement affected donation at the lowest educational levels. Cause of death influenced donation, with motor vehicle crash victims donating more often. The strongest factor in consent for donation was ethnicity; whites were more likely to donate than were other ethnic groups. The combination of gender, ethnicity, and cause of death improved the probability of determining a positive outcome to 63%. Demographic information on donors and nondonors can increase public and professional understanding as well as influence decision making to improve donation. PMID- 9726215 TI - Barriers to organ donation in the Jewish community. AB - It has long been recognized that members of the Jewish community generally do not sign organ donor cards or consent to the donation of the organs of their family members. In order to address this issue, the position of Jewish law on organ donation was examined and a sample of the Jewish population of Toronto was surveyed in an attempt to better understand the reasons for the observed reluctance to donate within this community. The results confirmed that the rate of signing organ donor cards was much lower in the Jewish community than in the general population, and although other reasons do exist, the major barrier to donation was a perception that Jewish law prohibits such action. The study of Jewish law revealed that organ donation is permitted and, in fact, encouraged by all branches of modern Judaism. Finally, in response to these results, a guide titled "Organ Donation: A Jewish Perspective" was compiled to help explain both the religious and medical aspects of organ donation for Jewish people and transplant personnel. PMID- 9726216 TI - A statewide assessment of attitudes, beliefs, and behaviors among blacks toward donation. AB - This study is part of a developing statewide campaign to increase donation in the black community. Focus groups were conducted to validate information obtained from a market research firm's telephone survey. Among those surveyed, 86.6% indicated they were in favor of donation; 13.4% indicated they were not. The most common reasons for opposition were religious issues and unfamiliarity with the donation process. Among those who were in favor of donation, 65% were some-what likely and very likely to donate; 74% were somewhat likely and very likely to give consent for donation of a loved one's organs. When asked where they would expect to learn about donation, respondents overwhelmingly chose the medical community. Respondents also listed cultural sensitivity as a criterion for choosing who would handle the donation request. A campaign to address identified issues has been developed and will be implemented statewide. PMID- 9726217 TI - Determining preferred educational methods for neurological surgery residents regarding organ donation. AB - Design and implementation of professional education, especially physician education, continues to challenge procurement professionals. At the request of the American Association of Neurological Surgeons and the Congress of Neurological Surgeons, the United Network for Organ Sharing undertook a project to develop educational materials for neurological surgery residents. A survey tool was developed and administered on site at 11 neurological surgery residency programs in the United States. The survey explored the types of learning environments, teaching methods, educational resources, and audiovisual aids that neurological surgery residents typically experience during their residency programs. In addition, the survey sought to uncover the residents' informational needs regarding organ and tissue donation presentations as well as their educational program preferences. Based on our findings, neurological surgery residents prefer presentations that are brief and to the point, that are easily understood, that require no reading, that contain limited important information, and that always include food. PMID- 9726218 TI - Analysis of organ donors in the peripartum period. AB - This study was a retrospective review of 252 brain-dead potential donors from 1990 to 1996, including 5 organ donors in the peripartum period. The purpose of the study was to determine the effects of pregnancy on organ donor management and recipient outcome. Case analysis of 5 pregnant donors identified problems with hemodynamic stability and electrolyte abnormalities, including hypernatremia, hyperchloremia, and hypocalcemia. In addition, blood glucose was frequently elevated. Two donors were treated for diabetes insipidus. All 5 donors produced organs for 20 transplant recipients. Five heart recipients (including 1 heart lung), 4 liver recipients, 4 kidney recipients, and 4 pancreas-kidney recipients have reported excellent outcomes. The use of organs from brain-dead organ donors in the peripartum period has minimal impact on donor management and recipient outcome. PMID- 9726219 TI - An in-house coordinator program to increase organ donation in public teaching hospitals. AB - The organ donor referral rates at Ben Taub General Hospital, a level 1 county trauma facility in Houston, Tex, were in the high 90s from 1993 through 1996. However, organs were procured from only 25% of those potential donors who were medically suitable. LifeGift Organ Donation Center identified two issues concerning the hospital's organ donor referrals: early medical management problems and a low consent rate. The hospital and LifeGift developed an in-house coordinator program along with specific protocols that have markedly increased the quantity of organ donors. In fiscal year 1994, 15 organ donors were recovered; in fiscal year 1995, following the implementation of the program, 25 organ donors were recovered. In fiscal year 1996, 39 organ donors were recovered. The in-house coordinators' consent rate was 72% of medically suitable potential donors. Early referrals for evaluation contributed to these successes. PMID- 9726220 TI - Hemodiabsorption in treatment of hepatic failure. AB - Conducted at the University of California, Los Angeles, this randomized, prospectively controlled study was designed to measure the effects of a hemodiabsorption device in the treatment of 11 patients with hepatic failure and stage III to IV encephalopathy. The results of 5 days of treatment with the hemodiabsorption device were compared with those of standard intensive care procedures. The BioLogic-DT System, a simple sorbent-based system, demonstrated a slight improvement in the neurologic status of patients, a significant improvement in the physiologic status of patients, and an improved outcome for treated patients as opposed to nontreated patients. PMID- 9726221 TI - Identifying donor concerns to increase live organ donation. AB - Increasing organ donation from live donors is a means to increase the pool of kidneys available for transplantation. To increase the number of live kidney donors, the major concerns of donors must be determined and addressed. This article describes the health and socioeconomic concerns of 61 live kidney donors from the Johns Hopkins Institutions between February 1, 1995 and December 1, 1997. Seventy-five percent of donors reported they had concerns, the most common ones being the effect of donation on their future health, how many work days they would miss, the ability to return to the same activities, and the pain they would experience. Other reported concerns consisted of fear of dying, the risks to a future pregnancy with one kidney, the fear that a son or daughter may need a kidney in the future, and the impact of donation on the family health insurance premium. Interventional strategies including the use of the laparoscopic donor nephrectomy procedure are offered to address these concerns. PMID- 9726222 TI - Pregnancy outcomes in 10 female pancreas-kidney recipients. AB - Female recipients of pancreas-kidney transplants may have an increased chance for pregnancy, because transplantation often restores fertility. Data on pregnancy after pancreas-kidney transplantation were analyzed by the National Transplantation Pregnancy Registry at US transplant centers. Ten recipients who were on cyclosporine-based immunosuppression were studied. A total of 15 pregnancies had resulted, of which 12 were live births. Among the 12 newborns, prematurity and low birth weight occurred in 75% and 83% of the cases, respectively. Three had complications associated with prematurity. Two thirds of the infants were delivered by cesarean section. All children are developing well with no apparent residual problems. During pregnancy, hypertension and urinary tract infections occurred frequently among recipients. Two recipients had three subsequent graft losses within 2 years of giving birth; however, both were successfully retransplanted. Successful pregnancy is possible for female pancreas kidney recipients. PMID- 9726223 TI - Religious outreach for organ and tissue donation. PMID- 9726224 TI - Dominance of cyclooxygenase-2 in the regulation of pancreatic islet prostaglandin synthesis. AB - Dramatic, scientifically important discoveries in prostaglandin (PG) pharmacology and physiology have taken place over the past decade. Chief among these discoveries is the identification of two separate forms of cyclooxygenase (COX), a constitutive and an inducible form, both of which exist in most tissues. The pancreatic islet is an exception to this rule because it continually and dominantly expresses the inducible form, COX-2. It has also been learned that nonsteroidal anti-inflammatory drugs affect the two forms of COX with different potencies, a finding with far-reaching clinical implications. An equally important finding is that PGE2, which is known to negatively modulate glucose induced insulin secretion, has at least four different subtypes of receptors with different mechanisms of action and metabolic consequences. These recent changes in our understanding of the molecular regulation of PG synthesis call for a reconsideration of previous hypotheses involving PGE2 as a regulator of beta-cell function in physiological and pathophysiological states. PMID- 9726226 TI - Expression of glucokinase in cultured human muscle cells confers insulin independent and glucose concentration-dependent increases in glucose disposal and storage. AB - Insulin resistance, as is found in skeletal muscle of individuals with obesity and NIDDM, appears to involve a reduced capacity of the hormone to stimulate glucose uptake and/or phosphorylation. The glucose phosphorylation step, as catalyzed by hexokinase II, has been described as rate limiting for glucose disposal in muscle, but overexpression of this enzyme under control of a muscle specific promoter in transgenic mice has had limited metabolic impact. In the current study, we investigated in a cultured muscle model whether expression of glucokinase, which in contrast to hexokinase II is not inhibited by glucose-6 phosphate (G-6-P), would have a pronounced metabolic impact. We used a recombinant adenovirus containing the cDNA-encoding rat liver glucokinase (AdCMV GKL) to increase the glucose phosphorylating activity in cultured human muscle cells by fourfold. G-6-P levels increased in AdCMV-GKL-treated cells in a glucose concentration-dependent manner over the range of 1-30 mmol/l, whereas the much smaller increases in G-6-P in control cells were maximal at glucose concentrations <5 mmol/l. Further, cells expressing glucokinase accumulated 17 times more 2-deoxyglucose-6-phosphate than control cells. In AdCMV-GKL-treated cells, the time-dependent rise in G-6-P correlated with an increase in the activity ratio of glycogen synthase. AdCMV-GKL-treated cells also exhibited a 2.5 to 3-fold increase in glycogen content and a four- to fivefold increase in glycolytic flux, proportional to the increase in glucose phosphorylating capacity. All of these observations were made in the absence of insulin. Thus we concluded that expression of glucokinase in cultured human muscle cells results in proportional increases in insulin-independent glucose disposal, and that muscle glucose storage and utilization becomes controlled in a glucose concentration-dependent manner in AdCMV-GKL-treated cells. These results encourage testing whether delivery of glucokinase to muscle in vivo has an impact on glycemic control, which could be a method for circumventing the failure of insulin to stimulate glucose uptake and/or phosphorylation in muscle normally in insulin-resistant subjects. PMID- 9726225 TI - Depot-related gene expression in human subcutaneous and omental adipocytes. AB - Human omental adipocytes display a range of biochemical properties that distinguish them from adipocytes of subcutaneous origin. However, information about site-related gene expression in human fat cells is limited. We have previously demonstrated that leptin mRNA is markedly overexpressed in abdominal subcutaneous (SC) compared with omental (Om) adipocytes. To further investigate depot-specific differences in adipocyte gene expression, we have measured, in paired samples of isolated human adipocytes obtained from SC and Om fat depots, the expression of mRNAs encoding a number of proteins involved in the control of adipocyte metabolism. In contrast to the marked site-related expression of leptin, genes encoding lipoprotein lipase (LPL), hormone-sensitive lipase (HSL), peroxisome proliferator-activated receptor-gamma (PPAR-gamma), tumor necrosis factor-alpha (TNF-alpha), and adipsin were not consistently differentially expressed. Of note, a highly significant inverse correlation between adipocyte PPAR-gamma expression and BMI (r = -0.7, P = 0.0005) was found. In parallel experiments, differential display was used in an attempt to identify novel and/or unexpected adipocyte genes that were expressed in a site-related manner. No transcript that was unique to one or another depot was found, but cellular inhibitor of apoptosis protein-2 (cIAP2) mRNA, which has not previously been reported in adipocytes, was expressed at higher levels in Om than SC adipocytes (Om > SC in all eight subjects; mean Om:SC ratio 1.9 +/- 0.2, P < 0.01). Because cIAP2 may be involved in the regulation of TNF-alpha signaling, this raises the possibility that depot-specific differences may exist in the regulation of adipocyte apoptosis. Thus, of the mRNAs examined to date, only leptin and cIAP2 show consistent site-related expression, suggesting that these molecules may have important roles in determining functional properties particular to individual adipose depots. Given the importance of PPAR-gamma in adipocyte development and insulin sensitivity, the inverse correlation between adipocyte PPAR-gamma mRNA levels and adiposity may represent a local regulatory mechanism restraining fat accumulation and/or may be related to the reduction of insulin sensitivity that occurs with increasing fat mass. PMID- 9726227 TI - Long-term survival of segmental pancreas isografts in NOD/Lt mice treated with anti-CD4 and anti-CD8 monoclonal antibodies. AB - Spontaneously diabetic nonobese diabetic (NOD/Lt) mice were treated with anti-T cell monoclonal antibodies (mAbs) at the time of grafting with vascularized segmental pancreas isografts. Recipients were either untreated or given anti-CD4 and/or anti-CD8 mAbs (0.5 mg/20-g mouse on each of 4 consecutive days), which reduced target cell levels to <5% of normal. Graft function was monitored by measuring blood glucose (BG) levels. Transplants were removed for histological examination when BG returned to >20 mmol/l for two consecutive readings. Isografts from 3- to 4-week-old prediabetic mice placed in untreated diabetic NOD mice ceased functioning in 9-13 days with a mean survival time (MST) +/- SD of 10 +/- 2. Treatment with anti-CD4 prolonged survival significantly (MST = 61 +/- 35 days, P < 0.05 compared with untreated control mice). Anti-CD8 treatment was less effective, but it still significantly improved graft survival (MST = 24 +/- 9 days, P < 0.05 compared with untreated control mice). Anti-CD8 plus anti-CD4 treatment was highly effective in inhibiting autoimmune destruction of the grafts (MST = 97 +/- 8 days). This clearly demonstrates that transient inactivation of most T-cells with anti-CD4 plus anti-CD8 mAbs effectively controls autoimmune disease in the isograft, despite recovery of CD4 and CD8 T-cells to normal levels. Although insulitis developed in the long-term grafts, insulitis scores did not increase between 33 and 100 days, and none of the mice progressed to IDDM in 100 days. Histology showed a predominantly peri-islet T-cell and macrophage infiltrate with ductal expression of the cytokines interleukin (IL)-4, IL-2, and interferon-gamma. There was little infiltrate or expression of cytokines within the islets. Thus, mAb treatment at the time of grafting allowed isograft survival and prevented progression from insulitis to beta-cell destruction. PMID- 9726228 TI - Expression of Gal alpha(1,3)gal on neonatal porcine islet beta-cells and susceptibility to human antibody/complement lysis. AB - Neonatal porcine pancreases may be a potential source of islets for transplantation into patients with type 1 diabetes; however, whether these cellular grafts will be susceptible to damage by human natural antibody-mediated rejection remains controversial. Although we and others have demonstrated that porcine islets bind human IgG and IgM, it remains unknown if they express the xenoreactive antigen Gal alpha(1,3)Gal beta(1,4)GlcNAc-R (Gal epitope). In this study, by using the Gal-specific lectin IB4 for immunohistochemistry and fluorescence-activated cell sorter (FACS) analysis, we determined which cell types present in porcine neonatal islet cell (NIC) aggregates express the Gal epitope and which ones are susceptible to lysis by activation of the human complement. After FACS analysis, 30.0 +/- 3.0% of porcine NICs were shown to express Gal, whereas 70.0 +/- 2.0% did not. Histological assessment of Gal expressing cells revealed that 54.9 +/- 8.8% stained positive for either insulin or glucagon. In contrast, 68.8 +/- 8.4% of the Gal-negative population stained positive for the pancreatic hormones insulin and glucagon. Incubation of either the Gal-positive or -negative cells with human AB serum plus complement for 1.5 h resulted in the lysis of >90% of the cells. These results demonstrate that porcine NIC aggregates are composed of Gal-expressing cells and that expression of Gal is not restricted to nonendocrine cells. Furthermore, both Gal-positive and Gal-negative cells are susceptible to human antibody/complement-mediated cytolysis, suggesting that this form of immunological destruction is an obstacle that will need to be overcome before porcine NIC aggregates can be used clinically. PMID- 9726229 TI - Tissue specificity of sulfonylureas: studies on cloned cardiac and beta-cell K(ATP) channels. AB - Sulfonylureas stimulate insulin secretion from pancreatic beta-cells by closing ATP-sensitive K+ (K(ATP)). The beta-cell and cardiac muscle K(ATP) channels have recently been cloned and shown to possess a common pore-forming subunit (Kir6.2) but different sulfonylurea receptor subunits (SUR1 and SUR2A, respectively). We examined the mechanism underlying the tissue specificity of the sulfonylureas tolbutamide and glibenclamide, and the benzamido-derivative meglitinide, using cloned beta-cell (Kir6.2/SUR1) and cardiac (Kir6.2/SUR2A) K(ATP) channels expressed in Xenopus oocytes. Tolbutamide inhibited Kir6.2/SUR1 (Ki approximately 5 micromol/l), but not Kir6.2/SUR2A, currents with high affinity. Meglitinide produced high-affinity inhibition of both Kir6.2/SUR1 and Kir6.2/SUR2A currents (Kis approximately 0.3 micromol/l and approximately 0.5 micromol/l, respectively). Glibenclamide also blocked Kir6.2/SUR1 and Kir6.2/SUR2A currents with high affinity (Kis approximately 4 nmol/l and approximately 27 nmol/l, respectively); however, only for cardiac-type K(ATP) channels was this block reversible. Physiological concentrations of MgADP (100 micromol/l) enhanced glibenclamide inhibition of Kir6.2/SUR1 currents but reduced that of Kir6.2/SUR2A currents. The results suggest that SUR1 may possess separate high-affinity binding sites for sulfonylurea and benzamido groups. SUR2A, however, either does not possess a binding site for the sulfonylurea group or is unable to translate the binding at this site into channel inhibition. Although MgADP reduces the inhibitory effect of glibenclamide on cardiac-type K(ATP) channels, drugs that bind to the common benzamido site have the potential to cause side effects on the heart. PMID- 9726230 TI - Functional analysis of a conditionally transformed pancreatic beta-cell line. AB - Development of beta-cell lines for cell therapy of diabetes is hindered by functional deviations of the replicating cells from the normal beta-cell phenotype. In a recently developed cell line, denoted betaTC-tet, derived from transgenic mice expressing the SV40 T antigen (Tag) under control of the tetracycline (Tc) gene regulatory system, growth arrest can be induced by shutting off Tag expression in the presence of Tc. Here, we compared differentiated cell functions in dividing and growth-arrested betaTC-tet cells, both in culture and in vivo. Proliferating cells stably maintained normal glucose responsiveness for >60 passages in culture. Growth-arrested cells survived for months in culture and in vivo and maintained normal insulin production and secretion. After growth arrest, the cells gradually increased their insulin content three- to fourfold. This occurred without significant changes in insulin biosynthetic rates. At high passage numbers, proliferating betaTC-tet cells exhibited an abnormal increase in hexokinase expression. However, the upregulation of hexokinase was reversible upon growth arrest. Growth-arrested cells transplanted intraperitoneally into syngeneic recipients responded to hyperglycemia by a significant increase in insulin secretion. These findings demonstrate that transformed beta-cells maintain function during long periods of growth arrest, suggesting that conditional transformation of beta-cells may be a useful approach for developing cell therapy for diabetes. PMID- 9726231 TI - Role of cAMP in upregulation of insulin secretion during the adaptation of islets of Langerhans to pregnancy. AB - Islets undergo a number of upregulatory changes to meet the increased demand for insulin during pregnancy, including an increase in glucose-stimulated insulin secretion with a reduction in the stimulation threshold. Treatment with the lactogenic hormone prolactin (PRL) in vitro has been shown to induce changes in islets similar to those observed during pregnancy. We examined cAMP production in islets treated with PRL to determine if changes in cAMP are involved in the upregulation of insulin secretion. Insulin secretion and cAMP concentrations were measured from islets in response to a suprathreshold (6.8 mmol/l) or high (16.8 mmol/l) glucose concentration in the presence of the phosphodiesterase inhibitor isobutylmethylxanthine. Insulin secretion increased by 2.1-, 5.0-, and 5.9-fold at the suprathreshold glucose concentration and by 1.6-, 2.3-, and 2.9-fold at the higher glucose concentration after 1, 3, and 5 days of PRL treatment, respectively. After a similar pattern, cAMP metabolism increased by 1.2-, 1.6-, and 2.1-fold at the suprathreshold glucose concentration and by 1.2-, 1.7-, and 2.2-fold at the high glucose concentration after 1, 3, and 5 days of PRL treatment, respectively. The similar increases in insulin secretion and cAMP concentration suggest that changes in cAMP metabolism are involved in lactogen induced upregulation of insulin secretion. To gain additional insight into the role of cAMP in the upregulation of islet function after lactogen treatment, we examined the relationship between changes in cAMP concentration and insulin secretion. Under all conditions (differing glucose concentrations and time periods), the increase in insulin release was directly proportional to the increase in cAMP. Thus increased glucose-stimulated insulin secretion from lactogen-treated islets could be accounted for by increased generation of cAMP and did not appear to require any further specific changes in intracellular processes mediated by cAMP. Because the PRL receptor is not directly involved in cAMP metabolism, the lactogen-induced increase in cAMP was most likely due to the increase in glucose metabolism that we have previously demonstrated in PRL treated islets and in islets during pregnancy. PMID- 9726232 TI - Ca2+-independent phospholipase A2 contributes to the insulinotropic action of cholecystokinin-8 in rat islets: dissociation from the mechanism of carbachol. AB - Insulin secretion induced by cholecystokinin-8 (CCK-8) was recently suggested to involve phospholipase A2 (PLA2) activation. In this study, we examined whether CCK-8 stimulates the Ca2+-independent form of PLA2 in isolated rat islets, in comparison with stimulation by the PLA2-activating cholinergic agonist carbachol. We found that CCK-8 (100 nmol/l; 5.6 mmol/l glucose) induces lysophosphatidylcholine accumulation from [3H]palmitate-prelabeled islets (170 +/ 39%; P = 0.003) as well as arachidonic acid (AA) efflux from [3H]AA-prelabeled islets (190 +/- 13%; P < 0.001), and that p-amylcinnamoylantranilic acid (ACA) (50 micromol/l)-mediated PLA2 inhibition reduces CCK-8-induced AA efflux (52 +/- 11%; P = 0.001) and insulin secretion (67 +/- 16%; P < 0.001). Neither the Ca2+ channel antagonist verapamil (100 micromol/l) nor the Ca2+ATPase inhibitor thapsigargin (1 micromol/l) affected CCK-8-induced AA efflux and insulin secretion. Furthermore, despite removal of extracellular Ca2+, CCK-8 still increased AA efflux (48 +/- 14%; P = 0.006) and insulin secretion (105 +/- 46%; P = 0.025). In contrast, carbachol (100 micromol/l)-stimulated AA efflux was reduced by verapamil by 36 +/- 6% (P < 0.001) and abolished by removal of extracellular Ca2+. Overnight protein kinase C (PKC) downregulation by 12-O tetradecanoyl phorbol-13-acetate (TPA) (500 nmol/l) reduced CCK-8-induced AA efflux (45 +/- 12%; P = 0.003) and insulin secretion (40 +/- 16%; P = 0.020). No additive action regarding either AA formation or insulin secretion was seen by combining TPA overnight and ACA, which implies the involvement of an additional PLA2- and PKC-independent signaling mechanism. The results show that CCK-8, in contrast to carbachol, activates Ca2+-independent PLA2 in islets and that the PLA2-activating capacity of CCK-8 is partly PKC dependent. Hence, Ca2+ independent PLA2 seems important for the insulinotropic effect of CCK-8, but not for that of carbachol. PMID- 9726233 TI - Blood-to-brain glucose transport and cerebral glucose metabolism are not reduced in poorly controlled type 1 diabetes. AB - To test the hypothesis that blood-to-brain glucose transport is reduced in poorly controlled type 1 diabetes, we studied seven patients with a mean (+/- SD) HbA1c level of 10.1 +/- 1.2% and nine nondiabetic subjects during hyperinsulinemic, mildly hypoglycemic (approximately 3.6 mmol/l, approximately 65 mg/dl) glucose clamps. Blood-to-brain glucose transport and cerebral glucose metabolism were calculated from rate constants derived from blood and brain time-activity curves- the latter determined by positron emission tomography (PET)--after intravenous injection of [1-(11)C]glucose using a model that includes a fourth rate constant to account for regional egress of 11C metabolites. Cerebral blood flow and cerebral blood volume were determined with intravenous H2(15)O and inhaled C(15)O, respectively, also by PET. At plateau plasma glucose concentrations of 3.6 +/- 0.0 and 3.7 +/- 0.1 mmol/l, rates of blood-to-brain glucose transport were similar in the two groups (23.7 +/- 2.2 and 21.6 +/- 2.9 micromol x 100 g( 1) x min(-1), P = 0.569, in the control subjects and the patients, respectively). There were also no differences in the rates of cerebral glucose metabolism (16.8 +/- 0.8 and 16.3 +/- 1.2 micromol x 100 g(-1) x min(-1), P = 0.693, respectively). Plasma epinephrine (1,380 +/- 340 vs. 450 +/- 170 pmol/l, P = 0.0440) and glucagon (26 +/- 5 vs. 12 +/- 1 pmol/l, P = 0.0300) responses to mild hypoglycemia were reduced in the patients with type 1 diabetes. We conclude that neither blood-to-brain glucose transport nor cerebral glucose metabolism is measurably reduced in people with poorly controlled type 1 diabetes. PMID- 9726234 TI - Alterations in skeletal muscle gene expression of ob/ob mice by mRNA differential display. AB - To identify molecules that contribute to insulin resistance, we compared the patterns of gene expression in skeletal muscle of the obese ob/ob mouse, a genetic model of obesity and severe insulin resistance, with that of its thin littermate (ob/+) using the mRNA differential display method. From about 9,000 cDNAs displayed, we found 12 differentially expressed in ob/ob mice skeletal muscle that could be recovered from the differential display gels and confirmed by Northern blot analysis and sequenced. Eight mRNAs were overexpressed in ob/ob muscle: Id2 (a negative regulator of the basic helix-loop-helix family of transcription factors), fast skeletal muscle troponin T, ribosomal protein L3, the integral protein of the peroxisomal membrane 22PMP, the mammalian homolog of geranylgeranyl pyrophosphate synthase, an mRNA related to phosphatidylinositol glycan-specific phospholipase D, and two unknown mRNAs. The level of overexpression of these mRNAs in skeletal muscle varied from a 500% increase to as little as a 25% increase. Two mRNAs were underexpressed 20-35%, including the f-subunit of mitochondrial ATP synthase and a retrovirus-related DNA. Two proteins with multiple transcripts, skeletal muscle alpha-tropomyosin and one for a repetitive sequence, showed a change in mRNA pattern of expression in the muscle of the ob/ob mouse. Because the primary genetic defect in the ob/ob mouse is known to be in the leptin gene, these data indicate how acquired alterations in gene expression of multiple classes of proteins may play a role in the complex pathogenesis of insulin resistance in obesity and diabetes. PMID- 9726235 TI - Glucose utilization and production in patients with maturity-onset diabetes of the young caused by a mutation of the hepatocyte nuclear factor-1alpha gene. AB - Mutations of the hepatocyte nuclear factor (HNF)-1alpha gene cause impaired insulin secretion and hyperglycemia in patients with maturity-onset diabetes of the young (MODY)3. Whether these mutations also affect glucose metabolism in tissues other than the beta-cell has not yet been documented. We therefore assessed, in five MODY3 patients and a dozen healthy control subjects, insulin secretion, oxidative and nonoxidative glucose disposal, and glucose production during a two-step hyperglycemic clamp and a euglycemic hyperinsulinemic (0.4 mU x kg(-1) x min(-1)) clamp. Compared with healthy control subjects, MODY3 patients had higher fasting plasma glucose (+100%) but similar rates of fasting glucose production and oxidation. Both the early and late phases of insulin secretion were virtually abolished during the hyperglycemic clamp, and glucose production was suppressed by only 43% in MODY3 patients vs. 100% in healthy control subjects. The rate of glucose infusion required to produce a 5 mmol/l increase above basal glycemia was reduced by 30%, net nonoxidative glucose disposal (which is equal to net glycogen deposition) was inhibited by 39%, and net carbohydrate oxidation during hyperglycemia was 25% lower in MODY3 patients compared with control subjects. Insulin-stimulated glucose utilization and oxidation measured during the hyperinsulinemic clamp (at approximately 200 pmol/l insulin) were identical in MODY3 patients and in healthy control subjects, indicating that peripheral insulin sensitivity was not altered. Suppression of endogenous glucose production was, however, mildly impaired. It is concluded that MODY3 patients have severely depressed glucose-induced insulin secretion. The development of hyperglycemia in these patients appears to be caused by a decreased stimulation of glucose utilization, oxidation, and nonoxidative glucose disposal as well as by a blunted suppression of endogenous glucose output. These phenomena are essentially secondary to insulinopenia, whereas insulin sensitivity remains intact. PMID- 9726236 TI - Mice expressing human but not murine beta3-adrenergic receptors under the control of human gene regulatory elements. AB - Beta-adrenergic receptors (ARs) are expressed predominantly in adipose tissue, and beta3-selective agonists are effective anti-obesity drugs in rodents. Rodent and human beta3-ARs differ with respect to expression in white versus brown adipocytes as well as their ability to be stimulated by beta3-AR-selective agonists. Humans express beta3-AR mRNA abundantly in brown but not white adipocytes, while rodents express beta3-AR mRNA abundantly in both sites. To determine the basis for this difference, we have transgenically introduced 74 kilobases (kb) of human beta3-AR genomic sequence into gene knockout mice lacking beta3-ARs. Importantly, human beta3-AR mRNA was expressed only in brown adipose tissue (BAT) of transgenic mice, with little or no expression being detected in white adipose tissue (WAT), liver, stomach, small intestine, skeletal muscle, and heart. This pattern of expression differed from that observed in mice bearing a murine beta3-AR genomic transgene in which beta3-AR mRNA was expressed in both WAT and BAT, but not in other sites. Furthermore, we have transgenically introduced smaller human constructs containing -14.5 and -0.6 kb of upstream sequence into beta3-AR gene knockout mice. Both -14.5 and -0.6 kb constructs were expressed in BAT but not WAT. Thus, human but not murine cis-regulatory elements direct beta3-AR gene expression preferentially to brown adipocytes. Identification of responsible cis-regulatory element(s) and relevant trans-acting factor(s) should provide insight into mechanisms controlling human beta3-AR gene expression. In addition, the beta3-AR agonist, CGP-12177, stimulated oxygen consumption in mice expressing human but not murine beta3-ARs by 91% compared with only 49% in control beta3-AR gene knockout mice, demonstrating that the human beta3-AR can functionally couple with energy expenditure. These "humanized" mice should assist us in the development of drugs that may become effective anti obesity agents in humans. PMID- 9726237 TI - Brief twice-weekly episodes of hypoglycemia reduce detection of clinical hypoglycemia in type 1 diabetes mellitus. AB - We tested the hypothesis that as few as two weekly brief episodes of superimposed hypoglycemia (i.e., doubling the average frequency of symptomatic hypoglycemia) would reduce physiological and behavioral defenses against developing hypoglycemia and reduce detection of clinical hypoglycemia in patients with type 1 diabetes mellitus (T1DM). Compared with nondiabetic controls, six patients with well-controlled T1DM (HbA1c, 7.5 +/- 0.7% [mean +/- SD]) exhibited absent glucagon responses and reduced epinephrine (P = 0.0027), norepinephrine (P = 0.0007), pancreatic polypeptide (P = 0.0030), and neurogenic symptom (P = 0.0451) responses to hypoglycemia as expected. In these patients, 2 h of induced hypoglycemia (50 mg/dl, 2.8 mmol/l) twice weekly for 1 month, compared in a random-sequence crossover design with an otherwise identical 2 h of induced hyperglycemia (150 mg/dl, 8.3 mmol/l) twice weekly for 1 month, further reduced the epinephrine (P = 0.0001) and pancreatic polypeptide (P = 0.0030) responses, tended to further reduce the norepinephrine and neurogenic symptom responses to hypoglycemia, and reduced cognitive dysfunction during hypoglycemia (P = 0.0271), all assessed in the investigational setting. In the clinical setting, induced hypoglycemia did not alter overall glycemic control, but did reduce the total number of symptomatic hypoglycemic episodes detected by the patients from 49 to 30 per month and lowered the mean +/- SE self-monitored blood glucose level during symptomatic hypoglycemia from 51 +/- 2 mg/dl (2.8 +/- 0.1 mmol/l) to 46 +/ 3 mg/dl (2.6 +/- 0.2 mmol/l) (P < 0.01). It also reduced the proportion of low regularly scheduled self-monitored values that were symptomatic by approximately 33%. Thus as little as doubling the frequency of symptomatic hypoglycemia further reduced both the key epinephrine response and clinical awareness of developing hypoglycemia, changes reasonably expected to increase the risk of severe iatrogenic hypoglycemia in T1DM. PMID- 9726238 TI - Hypoxia upregulates glucose transport activity through an adenosine-mediated increase of GLUT1 expression in retinal capillary endothelial cells. AB - Elevation of intracellular glucose within retinal vascular cells is believed to be an important causal factor in the development of diabetic retinopathy. The intracellular glucose concentration is regulated by both the rate of glucose metabolism and glucose transport. Because retinal hypoxia often precedes proliferative diabetic retinopathy, we have studied the regulation of the glucose transport system by hypoxia in cultured bovine retinal endothelial cells (BRECs). Because retinal ischemia is known to increase intracellular adenosine levels, which subsequently regulate hypoxia-inducible genes, such as vascular endothelial growth factor and erythropoietin, the role of adenosine and its receptor-mediated pathways has also been evaluated. Hypoxia (0.5% O2, 5% CO2, and 94.5% N2) stimulated GLUT1 mRNA expression in BRECs in a time-dependent manner with an 8.9 +/- 1.5-fold (P < 0.01) increase observed after 12 h. GLUT1 mRNA expression returned to baseline (1.4 +/- 0.3-fold of control) within 12 h after reinstitution of normoxia. N6-Cyclopentyl adenosine (adenosine A1 receptor agonist, Kd = 1 nmol/l) did not affect GLUT1 mRNA expression at concentrations up to 1 micromol/l, while 2-p-(2-carboxyethyl)-phenethyl-amino-5'-N ethylcarboxamidoadenosine and 5'-(N-ethylcalboxamido)-adenosine (adenosine A2 receptor [A2R] agonists, Kd = 15 and 16 nmol/l, respectively) increased mRNA levels at concentrations as low as 10 nmol/l. Maximal stimulation was 2.3 +/- 0.2 and 2.1 +/- 0.2-fold, respectively (P < 0.01). The adenosine A2a receptor antagonist 8-(3-chlorostyryl)caffeine (CSC) (Kd = 100 nmol/l for A2R) inhibited hypoxia-stimulated GLUT1 mRNA expression by 40 +/- 8% at 100 nmo/l. Hypoxia upregulated GLUT1 protein expression by 3.0 +/- 0.3-fold after 12 h (P < 0.01), but this response was attenuated by CSC (P < 0.05). Hypoxia increased glucose transport activity by 2.1 +/- 0.3-fold (P < 0.001) after 12 h, a response inhibited 65% by CSC (P < 0.01). A protein kinase A (PKA) inhibitor (H89, 20 micromol/l) suppressed hypoxia-induced GLUT1 mRNA expression by 42 +/- 9% (P < 0.01). These data suggest that hypoxia in BRECs upregulates glucose transport activity through an increase of GLUT1 expression that is partially mediated by adenosine, A2R, and the cAMP-PKA pathway. PMID- 9726239 TI - Intrauterine diabetes exposure and the risk of renal disease in diabetic Pima Indians. AB - The association between the diabetic intrauterine environment and renal disease was examined cross-sectionally in 503 Pima Indians with type 2 diabetes. Subjects were selected from participants in an ongoing study of diabetes and its complications in the Gila River Indian Community of Arizona. Subjects' exposure to diabetes in utero was established from periodic examinations conducted as part of the study. The prevalence of elevated urinary albumin excretion (UAE) (albumin to-creatinine ratio > or = 30 mg/g) was 40% (83 of 207) in the offspring of nondiabetic mothers, 43% (105 of 246) in the offspring of prediabetic mothers (i.e., women who were not diabetic at the time of the pregnancy but who developed diabetes after the pregnancy), and 58% (29 of 50) in the offspring of mothers who had diabetes during pregnancy. After controlling for age, sex, duration of diabetes, HbA1c, and mean arterial pressure in the offspring in a logistic regression analysis using generalized estimating equations, maternal diabetes during pregnancy was strongly associated with elevated UAE. The odds of elevated UAE in the offspring of mothers who had diabetes during pregnancy was 3.8 times (95% CI 1.7-8.4) that of the offspring of prediabetic mothers; the odds of elevated UAE in the offspring of nondiabetic and prediabetic mothers were similar (odds ratio of 0.94; 95% CI 0.59-1.5). We concluded that exposure to a diabetic intrauterine environment increases the risk of elevated UAE in diabetic Pima Indians. The effect of this exposure appears to be independent of other susceptibility factors that lead to nephropathy in diabetes. PMID- 9726240 TI - Differential effect of the antidiabetic thiazolidinediones troglitazone and pioglitazone on human platelet aggregation mechanism. AB - Troglitazone and pioglitazone, antidiabetic thiazolidinediones, are known to improve insulin resistance. However, the effect of these drugs on platelet aggregation remains unclear. The chemical structure of troglitazone contains vitamin E. Accordingly, we studied the effect of troglitazone, pioglitazone, and vitamin E on thrombin-induced platelet aggregation, metabolism of phosphoinositide, protein phosphorylation, protein kinase C (PKC)-alpha and beta, and phosphatidylinositol (PI) 3-kinase activation in vitro in human platelets. Maximum platelet aggregation by ADP, collagen, and thrombin decreased in the presence of 0.1-1 micromol/l troglitazone and 500 nmol/l vitamin E for 60 min compared with controls. However, pioglitazone did not inhibit ADP-, collagen , or thrombin-induced platelet aggregation. Pretreatment with troglitazone and vitamin E, but not with pioglitazone, resulted in decreases in thrombin-induced phosphatidic acid production, hydrolysis of phosphatidylinositol 4,5-bisphosphate by phospholipase C, and 47-kDa protein phosphorylation. Thrombin-induced PKC alpha and -beta activation in membrane fraction was suppressed by pretreatment with troglitazone and vitamin E, but not with pioglitazone. Separately, troglitazone and pioglitazone stimulated PI 3-kinase activity, but thrombin induced PI 3-kinase activation was suppressed by pretreatment with troglitazone and pioglitazone for 60 min. These results suggest that troglitazone and vitamin E, but not pioglitazone, have a potent inhibitory effect on platelet aggregation via suppression of the thrombin-induced activation of phosphoinositide signaling in human platelets. Finally, the chemical structure of vitamin E may contribute to the inhibitory effect of troglitazone on platelet aggregation in human platelets. PMID- 9726242 TI - Effect of angiotensin-converting enzyme (ACE) gene polymorphism on progression of renal disease and the influence of ACE inhibition in IDDM patients: findings from the EUCLID Randomized Controlled Trial. EURODIAB Controlled Trial of Lisinopril in IDDM. AB - We examined whether the ACE gene insertion/deletion (I/D) polymorphism modulates renal disease progression in IDDM and how ACE inhibitors influence this relationship. The EURODIAB Controlled Trial of Lisinopril in IDDM is a multicenter randomized placebo-controlled trial in 530 nonhypertensive, mainly normoalbuminuric IDDM patients aged 20-59 years. Albumin excretion rate (AER) was measured every 6 months for 2 years. Genotype distribution was 15% II, 58% ID, and 27% DD. Between genotypes, there were no differences in baseline characteristics or in changes in blood pressure and glycemic control throughout the trial. There was a significant interaction between the II and DD genotype groups and treatment on change in AER (P = 0.05). Patients with the II genotype showed the fastest rate of AER progression on placebo but had an enhanced response to lisinopril. AER at 2 years (adjusted for baseline AER) was 51.3% lower on lisinopril than placebo in the II genotype patients (95% CI, 15.7 to 71.8; P = 0.01), 14.8% in the ID group (-7.8 to 32.7; P = 0.2), and 7.7% in the DD group (-36.6 to 37.6; P = 0.7). Absolute differences in AER between placebo and lisinopril at 2 years were 8.1, 1.7, and 0.8 microg/min in the II, ID, and DD groups, respectively. The significant beneficial effect of lisinopril on AER in the II group persisted when adjusted for center, blood pressure, and glycemic control, and also for diastolic blood pressure at 1 month into the study. Progression from normoalbuminuria to microalbuminuria (lisinopril versus placebo) was 0.27 (0.03-2.26; P = 0.2) in the II group, and 1.30 (0.33-5.17; P = 0.7) in the DD group (P = 0.6 for interaction). Knowledge of ACE genotype may be of value in determining the likely impact of ACE inhibitor treatment. PMID- 9726241 TI - Reversed circadian blood pressure rhythm is associated with occurrences of both fatal and nonfatal vascular events in NIDDM subjects. AB - To assess the significance of reversed circadian blood pressure (BP) rhythms as a predictive factor of vascular events in NIDDM, vital status after an average 4 year follow-up was determined in 325 NIDDM subjects in whom the circadian BP profile had been monitored between 1988 and 1996. Circadian BP rhythm was analyzed by the COSINOR (a compound word for cosine and vector) method, as previously described. After exclusion of 37 dropped-out subjects, 288 were recruited to the further analysis, of which 201 had a normal circadian BP rhythm (group N) and the remaining 87 had a reversed one (group R). There was no difference in sex, HbA1c, the prevalence of smokers, serum lipids, or serum electrolytes between groups N and R at baseline, whereas age, the prevalence of hypertension, serum creatinine, and diabetic complications were more pronounced in group R than in group N. During the follow-up period (which averaged 52 months in group N and 36 months in group R), fatal and nonfatal vascular (cerebrovascular, cardiovascular, peripheral vascular arteries, and retinal artery) events occurred in 20 subjects in group N and 56 in group R. Unadjusted survival times and event-free times were estimated by the Kaplan-Meier product limit method, and there was a significant difference in both unadjusted survival and event-free survival rates between groups N and R (P < 0.001 each; log-rank test). The Cox proportional-hazards model adjusted for age, sex, circadian BP pattern, duration of diabetes, therapy for diabetes, various diabetic complications, and hypertension demonstrated that circadian BP pattern and age exhibited significant, high adjusted relative risks for fatal events, and that diabetic nephropathy, postural hypotension, and hypertension as well as circadian BP pattern exhibited significant, high adjusted relative risks with respect to the occurrence of various nonfatal vascular events. These results suggest that reversed circadian BP rhythm is associated with occurrences of both fatal and nonfatal vascular events in NIDDM subjects. PMID- 9726243 TI - Upregulation of osteopontin expression in renal cortex of streptozotocin-induced diabetic rats is mediated by bradykinin. AB - The model of streptozotocin (STZ)-induced diabetes in Wistar rats was used to study the expression of osteopontin during development of diabetic nephropathy. Diabetes was confirmed by serum glucose levels exceeding 16 mmol/l during the experimental period of 12 weeks. During this period of time, diabetic nephropathy developed, as characterized by a reduced glomerular filtration rate (2.7 +/- 0.3 ml/min in controls vs. 1.7 +/- 0.1 ml/min in diabetic rats) and proteinuria (8.3 +/- 1.7 mg/24 h in controls vs. 22.0 +/- 4 mg/24 h in diabetic rats). Northern blot analysis revealed a time-dependent upregulation of renal cortical osteopontin expression reaching 138 +/- 6% of control levels after 2 weeks and 290 +/- 30% (mean +/- SE, n = 6-9) after 12 weeks. By immunostaining, the increased osteopontin expression could be located to the tubular epithelium of the renal cortex. Chronic treatment of animals with ramipril (3 mg/kg) during the 12-week experimental period led to a further increase in osteopontin mRNA expression in diabetic animals, amounting to 570 +/- 73% (mean +/- SE, n = 6) of controls. Increased levels of osteopontin were not associated with accumulation of monocyte/macrophages that were identified by the cell type specific monoclonal antibody ED-1. The increased osteopontin expression in ramipril-pretreated rats was abolished by application of the bradykinin B2-receptor antagonist, icatibant (0.5 mg/kg). In addition, increased osteopontin expression in diabetic rats, which did not receive any treatment after STZ injection, could as well be reduced by icatibant given for the final 2 weeks of the experimental period. These data suggest that a strong bradykinin B2-receptor-mediated upregulation of osteopontin occurs during the pathogenesis of experimental diabetic nephropathy in rats. PMID- 9726244 TI - A common variant in PPP1R3 associated with insulin resistance and type 2 diabetes. AB - Selected candidate genes have been analyzed in the Pima Indians of Arizona based on evidence that insulin resistance and type 2 diabetes have significant genetic determinants. An amino acid substitution at codon 905 of the glycogen-targeting subunit of type 1 protein phosphatase that regulates skeletal muscle glycogenesis was recently reported to be associated with changes in insulin action in Danish subjects. In addition to the variant at 905, we report here a novel substitution at codon 883 and common variant of an "ATTTA" element in the 3'-untranslated region (UTR) of the corresponding gene (PPP1R3). The 3'-UTR variant resembled the mRNA-destabilizing AT(AU)-rich elements (AREs) and resulted in a 10-fold difference in reporter mRNA half-life, was correlated with PPP1R3 transcript and protein concentrations in vivo, and was associated with insulin resistance and type 2 diabetes in the Pimas. The variant is more common in Pimas (0.56) than in Caucasians (0.40). Because of its apparent effect on expression of PPP1R3, it may, in part, contribute to the higher prevalence of type 2 diabetes in this Native American population. PMID- 9726245 TI - Investigation of linkage of chromosome 8 to type 1 diabetes: multipoint analysis and exclusion mapping of human chromosome 8 in 593 affected sib-pair families from the U.K. and U.S. PMID- 9726246 TI - Uncoupling protein 3 is reduced in skeletal muscle of NIDDM patients. AB - Two recently described proteins in the mitochondrial uncoupling protein (UCP) family, UCP-2 and UCP-3, have been linked to phenotypes of obesity and NIDDM. We determined the mRNA levels of UCP-2 and UCP-3 in skeletal muscle of NIDDM patients and of healthy control subjects. No difference in the mRNA levels or in the protein expression of UCP-2 was observed between the two groups. In contrast, mRNA levels of UCP-3 were significantly reduced in skeletal muscle of NIDDM patients compared with control subjects. In the NIDDM patients, a positive correlation between UCP-3 expression and whole-body insulin-mediated glucose utilization rate was also noted. These results suggest that UCP-3 regulation may be altered in states of insulin resistance. PMID- 9726247 TI - Developmental regulation of LR11 expression in murine brain. AB - Receptors belonging to the low density lipoprotein receptor (LDLR) superfamily play important biological roles in addition to mediating lipoprotein metabolism. The recent discovery of a novel mosaic LDLR family member by us (Yamazaki H., Bujo, H., Kusunoki, J., Seimiya, K., Kanaki, T., Morisaki, N., Schneider, W.J., and Saito, Y. (1996) J. Biol. Chem. 271, 24761-24768) and others, which we termed LR11, offers the opportunity to gain new insights into receptor multifunctionality. The predominant expression of LR11 in brain and the presence of elements found in neural adhesion molecules suggested a function(s) in the central nervous system (CNS). In order to gain information about this complex receptor in an accessible system, we have molecularly characterized the murine LR11 and report on its detailed localization and developmental expression pattern. The primary sequence of the murine protein further establishes that LRlls are among the closest relatives within the LDLR family and that brain is the predominant site of expression. In situ hybridization showed that neuronal bodies such as Purkinje cells in the cerebellum and other neurons in the hippocampal formations and the cerebral cortex are particularly rich in LR11 transcripts. The developmental pattern of LR11 expression in brain, which peaks at 2 weeks, is in contrast to those of two other LDLR family members, the very low density lipoprotein receptor and the LDLR. During early development, murine LR11 expression levels are highly dependent on neural cell types. These findings are compatible with function(s) of LR11 in neural organization and, possibly, pathogenesis of degenerative brain diseases. In addition, detailed knowledge of LR11 biology will help to elucidate the roles of other mosaic proteins that share with LR11 elements whose function is not yet known. PMID- 9726248 TI - Mouse nicotinamide N-methyltransferase gene: molecular cloning, structural characterization, and chromosomal localization. AB - Nicotinamide N-methyltransferase (NNMT) catalyzes the N-methylation of nicotinamide and structurally related compounds. There are large strain-dependent variations in the expression of NNMT activity in mouse liver during growth and development, raising the possibility of developmental regulation of the gene. Therefore, we set out to clone and structurally characterize the mouse NNMT gene, Nnmt. The gene spanned approximately 16 kb and consisted of three exons, 348 bp, 208 bp, and 487 bp in length, with an initial 1228-bp intron and a second intron that was approximately 14 kb in length. The locations of the splice junctions within the gene were highly conserved compared with those in genes for structurally related methyltransferase enzymes. The Nnmt gene contained no canonical TATA box sequences, but an "initiator" (Inr) sequence was located at the site of transcription initiation as determined by 5' rapid amplification of cDNAs ends. A promoter was located within the initial 750 bp of the 5' flanking region of the gene according to studies of the expression of a reporter gene in HepG2 cells. 5'-Flanking region sequences for mouse strains with high and low hepatic NNMT activity differed with regard to a series of nucleotide substitutions, insertions, and deletions, with the most striking difference being a 12-bp insertion/deletion. The Nnmt gene mapped to mouse chromosome 9 in an area of conserved synteny to human chromosome 11q, consistent with the localization of the human NNMT gene to 11q23. Cloning and structural characterization of the mouse Nnmt gene will make it possible to study molecular genetic mechanisms involved in the expression of this important methyltransferase. PMID- 9726249 TI - CCAAT/enhancer binding proteins beta and delta regulate alpha1-acid glycoprotein gene expression in rat intestinal epithelial cells. AB - Isoforms of CCAAT/enhancer binding protein (C/EBP) are expressed in rodent intestine as well as in the rat intestinal epithelial cell line IEC-6. However, no specific roles have been attributed to these isoforms in intestinal epithelial cells. To determine whether C/EBP family members could be implicated in the regulation of acute-phase response gene expression in intestinal epithelial cells, we have studied the effect of glucocorticoids on expression of the alpha1 acid glycoprotein gene and C/EBP isoforms in IEC-6 cells. Glucocorticoids induced alpha1-acid glycoprotein gene expression in these cells. This induction coincided with an increase of DNA-binding capacity of both C/EBPbeta and C/EBPdelta to the B1 and B2 C/EBP-interacting sites localized in the rat AGP promoter. Transforming growth factor beta, (TGFbeta), a cytokine involved in the transcriptional regulation of several acute-phase plasma proteins, antagonized the glucocorticoid dependent induction of alpha1-acid glycoprotein gene expression. In parallel, TGFbeta downregulated the DNA-binding capacities of both the C/EBPbeta and C/EBPdelta isoforms. Mutations of the B1 or the B2 C/EBP-interacting site strongly reduced the responsiveness of the alpha1-acid glycoprotein promoter to glucocorticoids and TGFbeta. These results demonstrate a functional role for C/EBPbeta and C/EBPdelta in rat intestinal epithelial cells and suggest that these isoforms represent important modulators of the acute-phase response and of glucocorticoid, as well as TGFbeta, responsiveness. PMID- 9726250 TI - Mouse HNF-3/fork head homolog-1-like gene: structure, chromosomal location, and expression in adult and embryonic kidney. AB - Screening of a mouse kidney cDNA library with a HNF-3/fork head domain probe revealed cDNA Hfh-1L containing the highly conserved fork head DNA-binding domain. The Hfh1L cDNA shows 92.7% homology at the nucleic acid level with the fork head gene HFH-1 from rat. Southern blot analyses demonstrated that the Hfh 1L gene is highly conserved in a wide variety of species, including goldfish and frog. Sequencing the corresponding genomic clone, we found that the Hfh-1L gene is most likely intronless. By interspecific back-cross analysis, the Hfh-1L gene was localized to mouse chromosome 13. In order to analyze the expression pattern of Hfh-1L, we performed Northern blot analyses and revealed a 2.7-kb transcript in adult kidney and stomach. In situ hybridization experiments of adult mouse kidney showed Hfh-1L expression in the outer medulla of the kidney and the transitional epithelium. In light of the significance of a number of fork head genes in early embryonic development, the pattern of expression during murine embryogenesis was examined by reverse transcriptase-polymerase chain reaction (RT PCR), and Hfh-1L transcripts were detected in mouse embryos at every stage tested from day 10.5 to 16.5 postconception (p.c.) and in the developing metanephros of 14.5- and 15.5-day p.c. embryos. This expression pattern suggests that the Hfh-1L gene is involved in the development of the kidney. PMID- 9726251 TI - A composite regulatory element in the first intron of the estrogen-responsive very low density apolipoprotein II gene. AB - During periods of egg laying in the chicken, when circulating levels of estrogen are increased, the liver-specific estrogen-dependent very low density apolipoprotein II (apoVLDLII) gene is expressed. This expression takes place primarily at the level of transcription, driven by two estrogen response elements that reside in the promoter. In transient transfection assays, expression is increased fourfold when the first intron is added to the promoter construct, indicating that 75% of the regulation comes from intron A. Using in vitro DNase I footprinting, six protein-binding sites were revealed throughout the first intron. The functional significance of these binding sites was evaluated by mutation and transient transfection. Two of the protein-binding regions were shown to increase transcription. Site-specific mutations introduced at either the +66 to +86 or +112 to +129 sites disrupted trans-factor binding and reduced the estrogen-dependent expression by 45% and 34%, respectively. A plasmid containing both mutations resulted in a 43% decrease in expression, indicating that the contributions of these regions are not additive. Competition with known sequences in electrophoretic mobility shift assays suggested that the +66 to +86 site binds a chicken member of the nuclear receptor transcription factor family. PMID- 9726252 TI - Molecular cloning and expression of stromalin protein from Drosophila melanogaster: homologous to mammalian stromalin family of nuclear proteins. AB - We report the cloning of a new cDNA from Drosophila melanogaster that encodes an open reading frame of 1116 amino acid residues. It is the insect homolog of the previously reported stromalin (SA) family of nuclear proteins in mammals (Carramolino et al. [1997]. Gene 195, 151-159). Taking into account the identical domain present in all the SA family members characterized to date, we have carried out polymerase chain reaction (PCR) using degenerate oligonucleotides from the 5' and 3' ends of one of those regions of the molecule and cDNA from D. melanogaster embryos. We isolated the homologous domain of the putative Drosophila SA molecule (DSA). This cDNA fragment was used as a radiolabeled probe for screening a cDNA library from Drosophila embryos, and we have cloned a full length cDNA for the SA homolog from an insect. The protein shows a good degree of identity with the mammalian stromalins SA-1 and SA-2, with the N and C ends being the most divergent regions of the molecule. The mRNA coding for this protein shows a molecular size of about 3.7 kb by Northern blot analysis and is essentially expressed in embryonic stage. The in situ hybridization experiments indicate that the DSA messenger is expressed mainly in neurogenic territories in the embryonic development of Drosophila. The DSA protein has been cloned and expressed in a baculovirus system, and polyclonal antibodies were generated against the recombinant molecule. Western blot analysis using these antibodies detected a main band corresponding to about 120 kDa, principally in embryos. PMID- 9726253 TI - Modulation of 17alpha-hydroxylase/17,20-lyase activity of guinea pig cytochrome P450c17 by site-directed mutagenesis. AB - Microsomal cytochrome P450c17 (17a-hydroxylase/17,20-Lyase) catalyzes two reactions in the delta5 and delta4 pathways leading to the production of C19 steroids. Transient expression of human, bovine, porcine, rat, and mouse P450c17 cDNAs showed that the protein has 17alpha-hydroxylase and 17,20-Lyase activities, converting pregnenolone and progesterone into delta5- and delta4-Cl9 steroids, respectively, although the rat and mouse proteins have a preferential pathway toward the delta4 steroids. The guinea pig (gp) P450c17 shares 46% to 70% amino acid identity with the corresponding proteins of other species, and further characterization indicated that the guinea pig enzyme only converts progesterone to androstenedione. In this study, we have tried to identify amino acid(s) in the gpP450c17 that governs such a steroid specificity. Among the various mutants that we have created, change of the arginine (R) residue at position 200 to an asparagine (N) (R200N) in the gpP450c17 protein increased reactivity toward pregnenolone compared with the wild-type enzyme. Pregnenolone was converted into 17alpha-hydroxypregnenolone and dehydroepiandrosterone. However, this gain occurred at the expense of the 17,20-lyase activity toward 17alpha hydroxyprogesterone. The R200N mutation in the gpP450c17 protein introduced a potential N-linked glycosylation site (200Asn-X-Thr202); however, substitution of the Thr202 residue by an asparagine (R200N/T202N), which abolishes the site, did not change the preference of the gpP450c17 mutant for pregnenolone. Furthermore, introduction of a putative glycosylation site at amino acid 185 in the gpP450c17 enzyme did not alter substrate specificity. The properties of the amino acid were also investigated, and neither the charge nor the size of the sidechain elicited change in the substrate specificity of gpP450c17. Thus, our results demonstrate that the mutation of arginine to asparagine at position 200 changes the substrate specificity of the gpP450c17 enzyme. PMID- 9726254 TI - Usage of cryptic splice sites in citrullinemia fibroblasts suggests role of polyadenylation in splice-site selection during terminal exon definition. AB - Citrullinemia is a human genetic disease caused by a deficient argininosuccinate synthetase. In fibroblasts established from a citrullinemia patient with a mutation at the 3' splice site of the terminal intron of the gene, three cryptic 3' splice sites; i.e., SA1275, SA1636, and SA1663, residing on the terminal exon were activated. The usage of the cryptic sites showed a gradient, with the most downstream site having the highest usage; i.e., SA1663 > SA1636 > SA1275. However, when these cryptic sites were relocated to the internal exon, SA1636 was used the most. The splice-site strength of SA1636 was at least 10-fold higher than that of SA1663 in this situation. The results suggest that the preferential usage of SA1663 residing on the terminal exon may depend on its proximity to the poly(A) signal rather than on the strength of the splice site. Furthermore, when the strength of the downstream-most splice site increased, almost all the RNAs spliced to this site. However, in the presence of the wild-type splice site, all the RNAs were processed to the authentic site. Apparently, the selection of splice site can be revealed only when the sites being selected do not differ too much in their strength. By using a naturally occurring human mutant gene as a model, this study reveals that polyadenylation may play an important role in the selection of splice site during terminal exon definition. PMID- 9726255 TI - Cloning and functional analysis of C. elegans 7B2. AB - The neuroendocrine protein 7B2 is a binding protein for the prohormone convertase 2 (PC2) and is required for the intracellular conversion of proPC2 to active PC2. Both full-length 7B2 and its carboxy-terminal 31-residue peptide (CT peptide) are capable of potent inhibition of PC2; the 7B2 protein thus regulates both the biosynthesis and the activity of PC2. Vertebrate 7B2s are highly conserved (92% 97% homology), and thus, species comparison has not been informative in assessing the crucial protein domains responsible for bioactivity. We here report the cloning of the Caenorhabditis elegans 7B2 protein. Although weakly conserved with the vertebrate sequences (23% similarity with mouse 7B2), C. elegans 7B2 contains the signature PPNPCP motif as well as a highly conserved heptapeptide within the CT peptide. In in vitro assays, C. elegans 7B2 possessed significant inhibitory activity against recombinant vertebrate PC2 (IC50 130 nM), and in two functional tests, the amino-terminal domain of C. elegans 7B2 facilitated the activation of proPC2. We conclude that despite low amino acid conservation overall, both functional domains within 7B2 have been conserved between the C. elegans and the vertebrate proteins. PMID- 9726256 TI - Early 1H magnetic resonance spectroscopy of acute head injury: four cases. AB - In an attempt to examine in vivo the early metabolic consequences of severe acute head injury, 1H MRS was performed in four patients from 8 to 25 h (mean 15 h) following trauma. In three of these patients, decompressive surgery was performed 4-5 h prior to the MRS. High levels of lactate (area of lactate peak >50% of the mean areas of the NAA, choline-containing, and creatine-containing compound peaks) were found at 8 h posttrauma in the one patient who was not operated on and at 10 h posttrauma in one of the patients who underwent surgery. In the other two postoperative patients, at 18 and 25 h after trauma, lactate levels were found to be low (lactate peak <20% of the mean area of the other three peaks). In the one patient who had a follow-up at 6 days and who had the largest initial lactate levels, these remained high. These findings suggest that high levels of lactate may not be an inevitable consequence of severe head injury and that similar MRS studies should be performed on each individual patient before therapies to reduce lactate are considered. There appeared to be no correlation between the relative amounts of lactate and outcome. PMID- 9726257 TI - Structured interviews for the Glasgow Outcome Scale and the extended Glasgow Outcome Scale: guidelines for their use. AB - The Glasgow Outcome Scale (GOS) is the most widely used outcome measure after traumatic brain injury, but it is increasingly recognized to have important limitations. It is proposed that shortcomings of the GOS can be addressed by adopting a standard format for the interview used to assign outcome. A set of guidelines are outlined that are directed at the main problems encountered in applying the GOS. The guidelines cover the general principles underlying the use of the GOS and common practical problems of applying the scale. Structured interview schedules are described for both the five-point GOS and an extended eight-point GOS (GOSE). An interrater reliability study of the structured interviews for the GOS and GOSE yielded weighted kappa values of 0.89 and 0.85, respectively. It is concluded that assessment of the GOS using a standard format with a written protocol is practical and reliable. PMID- 9726258 TI - Analyzing outcome of treatment of severe head injury: a review and update on advancing the use of the Glasgow Outcome Scale. AB - The Glasgow Outcome Scale (GOS), two decades after its description, remains the most widely used method of analyzing outcome in series of severely head-injured patients. This review considers limitations recognized in the use of the GOS and discusses a new approach to assessment, using a structured questionnaire-based interview. Assignments can be made to an extended eight-point scale (GOSE) as well as the original five-point approach-in each case, with a high degree of interobserver consistency. The assignments are coherent with the principles of the World Health Organization classification of impairments, disabilities, and handicaps, and their validity is supported by strong associations with the results of neuropsychological testing and assessment of general health status. The need to allow for disability existing before injury, issues concerning the time of assessment after injury, and subdivisions of the scale into "favorable" and "unfavorable" categories are discussed. It is concluded that, in its improved structured format, the Glasgow Outcome Scale should remain the primary method of assessing outcome in trials of the management of severe head injury. PMID- 9726260 TI - Mild traumatic brain injury does not modify the cerebral blood flow profile of secondary forebrain ischemia in Wistar rats. AB - The rat hippocampus is hypersensitive to secondary cerebral ischemia after mild traumatic brain injury (TBI). An unconfirmed assumption in previous studies of mild TBI followed by forebrain ischemia has been that antecedent TBI did not alter cerebral blood flow (CBF) dynamics in response to secondary ischemia. Using laser Doppler flowmetry (LDF), relative changes in regional hippocampal CA1 blood flow (hCBF) were recorded continuously to quantitatively characterize hCBF before, during, and after 6 min of forebrain ischemia in either normal or mildly traumatized rats. Two experimental groups of fasted male Wistar rats were compared. Group 1 (n = 6) rats were given 6 minutes of transient forebrain ischemia using bilateral carotid clamping and hemorrhagic hypotension. Group 2 (n = 6) rats were subjected to mild (0.8 atm) fluid percussion TBI followed 1 h after trauma by 6 min of transient forebrain ischemia. The laser Doppler flow probe was inserted stereotactically to measure CA1 blood flow. The electroencephalogram (EEG) was continuously recorded. During the forebrain ischemic insult there were no intergroup differences in the magnitude or duration of the decrease in CBF in CA1. In both groups, CBF returned to preischemic values within one minute of reperfusion but traumatized rats had no initial hyperemia. There were no intergroup differences in the CBF threshold when the EEG became isoelectric. These data suggest that the ischemic insult was comparable either with or without antecedent TBI in this model. This confirms that this model of TBI followed by forebrain ischemia is well suited for evaluating changes in the sensitivity of CA1 neurons to cerebral ischemia rather than assessing differences in relative ischemia. PMID- 9726259 TI - Sustained sensory/motor and cognitive deficits with neuronal apoptosis following controlled cortical impact brain injury in the mouse. AB - A mouse model of traumatic brain injury was developed using a device that produces controlled cortical impact (CCI), permitting independent manipulation of tissue deformation and impact velocity. The left parietotemporal cortex was subjected to CCI [1 mm tissue deformation and 4.5 m/s tip velocity (mild), or 6.0 m/s (moderate)] or sham surgery. Injured animals showed delayed recovery of pedal withdrawal and righting reflexes compared to sham-operated controls. Significant severity-related deficits in forepaw contraflexion and performance on a rotarod device were evident for up to 7 days. Using a beam walking task to measure fine motor coordination, pronounced deficits were apparent for at least 2 and 4 weeks following mild and moderate CCI, respectively. Cognitive function was evaluated using the water maze. Impairment of place learning, related to injury severity, was observed in mice trained 7-10 days following CCI. Similarly, working memory deficits were evident in a variation of this task when examined 21-23 days postinjury. Mild CCI caused necrosis of subcortical white matter with minimal damage to somatosensory cortex. Moderate CCI produced extensive cortical and subcortical white matter damage. Triple fluorescence labeling with terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL), antineuronal nuclear protein (NeuN), and Hoechst 33258 of parallel sections showed frequent apoptotic neurons. These findings demonstrate sustained and reproducible deficits in sensory/motor function and spatial learning in the CCI injured mouse correlating with injury severity. Mechanisms of neuronal cell death after trauma as well as strategies for evaluating novel pharmacological treatment strategies may be identified using this model. PMID- 9726261 TI - Effects of moderate, central fluid percussion traumatic brain injury on nitric oxide synthase activity in rats. AB - Experimental traumatic brain injury (TBI) damages cerebral vascular endothelium and reduces cerebral blood flow (CBF). The nitric oxide synthase (NOS) substrate, L-arginine, prevents CBF reductions after TBI, but the mechanism is not known. This study examined the possibility that post-traumatic hypoperfusion is due to reductions in the substrate sensitivity of NOS which are overcome by L-arginine. Isoflurane-anesthetized rats were prepared for TBI (midline fluid-percussion, 2.2 atm), sham-TBI, or no surgery (control), and were decapitated 30 min after injury or sham injury. The brains were removed and homogenized or minced for measurements of crude soluble or cell-dependent stimulated NOS activity, respectively. Baseline arterial oxygen, carbon dioxide, pH, or hemoglobin levels did not differ among control, sham, or TBI groups. Total cortical soluble NOS activity in TBI-treated rats was not significantly different from either untreated or sham groups when 0.45 microM or 1.5 microM L-arginine was added. Also, there were no differences in cell-dependent NOS activity among the three groups stimulated by 300 microM N-methyl-D-aspartate, 50 mM K+, or 10 microM ionomycin. These data suggest that TBI reduces CBF by a mechanism other than altering the substrate specificity or activation of nNOS. PMID- 9726262 TI - Endothelial-mediated dilations following severe controlled cortical impact injury in the rat middle cerebral artery. AB - The mechanisms associated with dysfunction of the cerebral vasculature following head trauma have not yet been fully elucidated. In an attempt to shed more light on the matter, we investigated the endothelial-mediated dilations in the rat middle cerebral artery (MCA) following severe traumatic brain injury (TBI). Rats were subjected to severe controlled cortical impact injury (CCI; 5 m/s, 130 ms duration, 3 mm deformation) over the right parietal cortex. At 24 h postinjury, ipsilateral segments of MCA and corresponding contralateral segments were isolated, mounted in a vessel chamber, and pressurized. The responses to 2 methylthio-ATP (2MeSATP), a selective agonist for the P2Y1 purinoceptors, N(omega)-nitro-L-arginine (L-NAME), an NO synthase inhibitor, and S-nitroso-N acetylpenicillamine (SNAP), an exogenous NO donor, were determined. 2MeSATP elicited concentration dependent dilations in all MCAs studied. Ipsilateral MCAs harvested following TBI or sham-TBI, showed similar maximum dilations to 2MeSATP [70 +/- 4% (n = 17) and 72 +/- 6% (n = 13), respectively]. However, TBI reduced the concentration of 2MeSATP necessary to elicit one-half of the maximum dilation (EC50) from 15 to 9 nM (p < 0.05). Inhibition of NO synthase with 10(-5) M L-NAME abolished the dilation to 2MeSATP in both TBI and sham-TBI MCAs. The constriction to L-NAME was significantly reduced in TBI MCAs compared to sham vessels. Dilations to SNAP, an NO donor, were not altered by TBI indicating that the mechanisms of dilation involving NO in the vascular smooth muscle were not affected. Unlike other pathological conditions which often diminish endothelial mediated responses, severe TBI enhanced the sensitivity to 2MeSATP without altering the maximum response. PMID- 9726263 TI - Toxic effect of hemoglobin on spinal cord neurons in culture. AB - The vulnerability of spinal cord neurons to hemoglobin was quantitatively assessed in primary cultures derived from fetal mice. Exposure to hemoglobin for 28 h in a serum-free medium resulted in concentration-dependent neuronal death, with an EC50 of 0.9 microM; glia were not injured. Neuronal death was decreased by the ferric iron chelator deferoxamine, the alpha-tocopherol analogue Trolox C, ascorbate, and exogenous catalase, but was potentiated by superoxide dismutase. Neuronal death was also increased by depletion of cellular glutathione with the gamma-glutamylcysteine synthetase inhibitor buthionine sulfoxamine; inhibition of endogenous catalase with 3-amino-1,2,4-triazole had no significant effect. These results suggest that hemoglobin is toxic to spinal neurons via an iron-dependent, oxidative mechanism involving a hydrogen peroxide intermediate, and support the hypothesis that hemoglobin release may contribute to neuronal loss after spinal cord trauma. PMID- 9726264 TI - Participant selection biases in a randomized clinical trial of alcoholism treatment settings and intensities. AB - Eligible participants and decliners in a randomized study of inpatient, intense outpatient, and standard outpatient treatments for alcoholics were compared and contrasted on a series of demographic, social stability, psychological, legal, drug use, problem severity, and treatment history variables. Among 302 individuals meeting eligibility requirements, those agreeing to participate, compared with decliners, were more likely to be unemployed, be residentially less stable, have legal problems, use other drugs, have a more severe alcohol problem, have a recent treatment history, and were less likely to have problems with violence. Participants also were more likely to be male and non-white, although gender and racial effects were not significant when other variables were controlled for. The implications of these findings for generalizing the results of inpatient-outpatient studies are discussed, and the need for routine reporting of decliner characteristics in research reports is stressed. PMID- 9726265 TI - The Alcoholics Anonymous Affiliation Scale: development, reliability, and norms for diverse treated and untreated populations. AB - Affiliation with Alcoholics Anonymous (AA) is an important variable to measure in many clinical and research activities. This paper reports on the development of an AA affiliation scale, and demonstrates its utility in a sample of 927 alcohol treatment seekers and 674 untreated problem drinkers. The scale is short (9 items), covers a range of AA experiences, and is internally consistent across diverse demographic groups, multiple health services settings, and treated and untreated populations. The validity of the scale is supported by the findings that treatment seekers report significantly higher AA affiliation than do untreated problem drinkers, and inpatients report higher affiliation than outpatients. Potential clinical and research applications of the scale are proposed. PMID- 9726267 TI - A critical evaluation of several alcohol screening instruments using the CIDI-SAM as a criterion measure. AB - Four alcohol screening instruments (the AUDIT, CAGE, MAST, and Svanum's scale) were administered to a sample of 306 undergraduate students at a Midwestern university and were compared with regard to several test characteristics, using the alcohol section of the CIDI-SAM (DSM-IV version) as the criterion measure. The performance of these instruments was evaluated using two subsets of subjects: (1) students who currently met diagnostic criteria for alcohol dependence (n = 35); and (2) students who met diagnostic criteria for alcohol dependence in the past and/or at present (i.e., lifetime diagnosis; n = 50). The AUDIT performed significantly better than the other three instruments in identifying students who were currently alcohol dependent, providing a moderate degree of clinical utility with this group. The four instruments did not differ significantly in their ability to identify students with a lifetime diagnosis; each measure provided only a modest degree of clinical utility with this group. PMID- 9726266 TI - How do physicians define "light," "moderate," and "heavy" drinking? AB - Although widely used, terms associated with consumption of alcohol--such as "light," "moderate," and "heavy"--are unstandardized. Physicians conveying health messages using these terms therefore may impart confusing information to their patients or to other physicians. As an initial attempt to assess if informal standardization exists for these terms, the present study surveyed physicians for their definitions of such terms. Physicians operationally defined "light" drinking as 1.2 drinks/day, "moderate" drinking as 2.2 drinks/day, and "heavy" drinking as 3.5 drinks/day. Abusive drinking was defined as 5.4 drinks/day. There was considerable agreement for these operational definitions, indicating there is indeed an informal consensus among physicians as to what they mean by these terms. Gender and age did not influence these definitions, but self-reported drinking on the part of physicians was a factor. We also asked physicians for their opinions regarding the effects of "light," "moderate," and "heavy" drinking on health in general and specifically on health-related implications for pregnant women, and whether they felt their patients shared these beliefs. PMID- 9726268 TI - Moderate alcohol consumption and estrogen levels in postmenopausal women: a review. AB - This report reviews the literature to evaluate association between moderate alcohol consumption and estrogen levels in healthy postmenopausal women. Of the eight studies available in literature on postmenopausal women who were not on estrogen therapy, two analyzed urine samples and six analyzed blood samples for estrogen levels. Of the two urine sample studies, only one reported positive association (p < 0.05) between alcohol consumption and estrogen (estrone and estradiol) levels that increased by 16 to 20%. Of the six blood sample studies, only two--one in American women and one in European women--reported significant increases (p < 0.05) in estradiol levels in response to alcohol consumption. In the American women study, estradiol levels increased only with wine and not with beer or whiskey. In the European women study, estradiol levels increased in Danish and Portuguese women, but not in Spanish women. Thus, further studies are required to establish correlation between moderate alcohol consumption and estrogen levels in postmenopausal women. Of the two studies on postmenopausal women who were on estrogen replacement therapy, one administered estradiol through transdermal patch (0.15 mg) and one orally (1 mg/day). In both studies, blood estradiol levels were measured after administering a single dose of ethanol orally (0.7-0.75 g/kg of body weight). Estradiol levels were increased by 22 and 300% in the transdermal patch and oral studies, respectively. These results suggest that alcohol consumption may increase blood estradiol levels in postmenopausal women who are on estrogen replacement therapy, and this may increase the risk of breast cancer. PMID- 9726269 TI - Effects of moderate alcohol consumption on the central nervous system. AB - The concept of moderate consumption of ethanol (beverage alcohol) has evolved over time from considering this level of intake to be nonintoxicating and noninjurious, to encompassing levels defined as "statistically" normal in particular populations, and the public health-driven concepts that define moderate drinking as the level corresponding to the lowest overall rate of morbidity or mortality in a population. The various approaches to defining moderate consumption of ethanol provide for a range of intakes that can result in blood ethanol concentrations ranging from 5 to 6 mg/dl, to levels of over 90 mg/dl (i.e., approximately 20 mM). This review summarizes available information regarding the effects of moderate consumption of ethanol on the adult and the developing nervous systems. The metabolism of ethanol in the human is reviewed to allow for proper appreciation of the important variables that interact to influence the level of exposure of the brain to ethanol once ethanol is orally consumed. At the neurochemical level, the moderate consumption of ethanol selectively affects the function of GABA, glutamatergic, serotonergic, dopaminergic, cholinergic, and opioid neuronal systems. Ethanol can affect these systems directly, and/or the interactions between and among these systems become important in the expression of ethanol's actions. The behavioral consequences of ethanol's actions on brain neurochemistry, and the neurochemical effects themselves, are very much dose- and time-related, and the collage of ethanol's actions can change significantly even on the rising and falling phases of the blood ethanol curve. The behavioral effects of moderate ethanol intake can encompass events that the human or other animal can perceive as reinforcing through either positive (e.g., pleasurable, activating) or negative (e.g., anxiolysis, stress reduction) reinforcement mechanisms. Genetic factors and gender play an important role in the metabolism and behavioral actions of ethanol, and doses of ethanol producing pleasurable feelings, activation, and reduction of anxiety in some humans/animals can have aversive, sedative, or no effect in others. Research on the cognitive effects of acute and chronic moderate intake of ethanol is reviewed, and although a number of studies have noted a measurable diminution in neuropsychologic parameters in habitual consumers of moderate amounts of ethanol, others have not found such changes. Recent studies have also noted some positive effects of moderate ethanol consumption on cognitive performance in the aging human. The moderate consumption of ethanol by pregnant women can have significant consequences on the developing nervous system of the fetus. Consumption of ethanol during pregnancy at levels considered to be in the moderate range can generate fetal alcohol effects (behavioral, cognitive anomalies) in the offspring. A number of factors--including gestational period, the periodicity of the mother's drinking, genetic factors, etc.--play important roles in determining the effect of ethanol on the developing central nervous system. A series of recommendations for future research endeavors, at all levels, is included with this review as part of the assessment of the effects of moderate ethanol consumption on the central nervous system. PMID- 9726270 TI - Predicting relapse to substance abuse as a function of personality dimensions. AB - The goal of the present study was to determine whether personality traits are related to return to heavy drinking or drug use following treatment for substance abuse. Personality characteristics of one hundred and eight patients residing on an inpatient substance abuse treatment program were assessed. Personality traits were examined using the 5-factor model of personality as measured by the NEO Personality Inventory. These patients were then followed for 1 year after discharge from the treatment program. These substance abuse patients scored higher than the NEO-Personality Inventory normative sample on the personality domains of Neuroticism and Conscientiousness. A survival analysis showed that Neuroticism and Conscientiousness from the NEO-Personality Inventory were significant predictors of relapse. Odds ratios showed that the risk of relapsing was greatest for those patients who were both low in conscientiousness and high in neuroticism. The relevance of these two broad personality dimensions to the development and maintenance of addiction is discussed. Treatment implications for patients who possess these personality risk factors are outlined. PMID- 9726271 TI - Self-reported flushing and genotypes of ALDH2, ADH2, and ADH3 among Taiwanese Han. AB - The aims of this study are to investigate whether self-reported facial flushing postalcohol consumption (PAC) among subjects with ALDH2*1/*1 can be attributed to ADH2 or ADH3 and whether the prediction of ALDH2 genotype can be improved by examining the combination of flushing and other accompanying reactions of PAC sensitivity. Fifty-eight subjects of Han ancestry in Taiwan were interviewed for alcohol-sensitivity reactions and their blood samples were genotyped for ALDH2, ADH2, and ADH3. For subjects with ALDH2*1/*1 (n = 46), 70% reported to have no flushing PAC and 30% reported flushing PAC. When subjects with ALDH2*1/*1 had ADH2*1/*1 (n = 11), all reported to have no flushing; otherwise, 35% (for ADH2*1/*2, n = 17) and 44% (for ADH2*2/*2, n = 18) reported flushing. For subjects with ALDH2*1/*1 and at least one ADH2*2 allele, the genotype of ADH3 was not associated with self-reported flushing. PAC flushers with ALDH2*1/*1 (50%) were more likely to report nausea than those with ALDH2*1/*2 (8%). The probability of ALDH2*1/*1 given flushing reported was 0.29, while the probability of ALDH2*1/*1 given both flushing and nausea reported was 0.71. The results indicate that self-reported flushing is determined by both ALDH2 and ADH2 and that prediction of ALDH2 genotype on the basis of self-reported flushing and nausea can help identify subjects at increased risk for alcoholism. PMID- 9726272 TI - Does carbohydrate-deficient transferrin predict the severity of alcohol withdrawal syndrome? AB - Alcohol withdrawal often causes severe complications. However, many addicts deny any abuse. Thus, the diagnosis of alcohol abuse frequently becomes difficult. Laboratory parameters are often used to support the diagnosis of alcohol abuse. Furthermore, laboratory parameters should facilitate the prediction of the severity of alcohol withdrawal syndrome (AWS). The most promising laboratory parameter indicating a recent elevated alcohol consumption is carbohydrate deficient transferrin (CDT). The aim of this study was to examine whether the measurement of CDT at admission can indicate a higher risk for the development of a complicated AWS. The severity of AWS was assessed by the AWS scale, consisting of two subscales for somatic and mental symptoms. CDT was measured by different methods (radioimmunoassay and HPLC). The radioimmunoassay for CDT (CDTect) yielded the best prediction. Our results showed a weak correlation between CDTect and the severity of AWS. However, there were great gender differences. In men, CDTect had the highest positive predictive value for a severe AWS (86.7%), whereas in women mean corpuscular volume was the best predictor (77.8%). However, the sensitivity of CDTect in men (25.5%), as well as mean corpuscular volume in females (29.2%), was too low for a screening test in a general hospital. PMID- 9726273 TI - An initial study of the relationship between platelet adenylyl cyclase activity and alcohol use disorder criteria. AB - Low platelet adenylyl cyclase (AC) activity has been previously proposed to be a trait marker reflecting a genetic predisposition to alcohol dependence. To determine whether low platelet AC activity in alcohol-dependent subjects may be related to specific diagnostic criteria of DSM-IV and ICD-10 alcohol use disorders, we analyzed responses obtained in structured clinical interviews of 36 subjects who were determined to be alcohol-dependent Platelet AC activity when stimulated by guanylyl-imidodiphosphate [Gpp(NH)p] or forskolin was significantly lower in alcohol-dependent subjects as a group, compared with controls. When we analyzed the responses of the alcohol-dependent subjects to questions used to establish the diagnosis of alcohol abuse/dependence and dichotomized the subjects by positive or negative responses, we found that Gpp(NH)p- and forskolin stimulated platelet AC activities were significantly lower among those alcohol dependent subjects who had positive responses to questions related to drinking despite negative effects on mood ("Did you ever continue to drink even though you knew it was making you feel depressed, uninterested in things, or suspicious or distrustful of other people?"), drinking despite negative effects on health ("Did you ever continue to drink even though you knew it was causing you a health problem or making a health problem worse?"), or violence when drinking ("Did you get into physical fights while drinking or right after drinking?"). The alcohol dependent subjects who had negative responses to these questions exhibited Gpp(NH)p- and forskolin-stimulated platelet AC activity that did not differ significantly from values in control subjects. The DSM-IV diagnosis of antisocial personality disorder did not distinguish alcohol-dependent subjects with regard to platelet AC activity. Gpp(NH)p and forskolin-stimulated AC activity may distinguish certain subtypes of alcoholics (i.e., those who develop negative mood in response to drinking, those who continue drinking despite health effects, and those who become violent while drinking). PMID- 9726275 TI - Voices of the afflicted. AB - Over the past 10 years, I have been privileged to conduct educational forums for audiences containing many recovering alcoholics or otherwise chemically dependent persons. In these forums about the addictive diseases and their treatment and research possibilities, significant interaction with the audience members occurs. During these interactions, certain anecdotal phenomena seem to predominate. The repetitive nature of these reports suggests the need for systematic investigation. As with editorial comments in major medical journals, observed phenomena and unanswered questions from those afflicted can be valuable in the generation of testable hypotheses. Perhaps the ideas presented herein will be useful in the development of future research on alcohol abuse and alcohol dependence. PMID- 9726274 TI - Effects of cues on memory in alcoholics and controls. AB - This study was conducted to determine whether alcoholics' memory difficulties are due, in part, to access (retrieval) or to availability (retention) deficits. Forty-four alcoholics (n = 20 females) and 44 controls (n = 22 females) learned a paired associate list consisting of 12 adjective-CCC trigram pairs. Half of the subjects in each group learned the list to a low degree of learning (DOL; 4/12 pairs); the remainder to a high DOL (8/12 pairs). Two distinct environmental contexts (providing implicit cues) were used during acquisition. Subjects then completed a cued recall (an explicit cue) test in either the same or a different room. Alcoholics were significantly inferior in the acquisition phase on trials required to reach criterion, regardless of DOL required [F(1,68) = 10.92, p = 0.002]. The main effect for implicit cuing was not significant; similarly, there were no significant interactions. In contrast, the explicit cue manipulation produced a significant group x DOL interaction on the number of trigrams correctly recalled [F = (1,77) = 6.38, p = 0.01]; alcoholics' recall did not benefit from the higher DOL in contrast to a significant improvement in recall by controls. The failure of alcoholics to demonstrate improvement with higher levels of learning is consistent with a deficit in the availability of information. The results confirm previous reports of recovering alcoholics' verbal learning and memory dysfunction, and suggest that these deficits may be attributed, in part, to a deficit in the availability of information (retention). PMID- 9726276 TI - Investigation into the use of alcohol at thermal spas: can a spa represent a place for primary prevention? AB - Primary prevention is one of the most effective means used nowadays to make the general population aware of most of the problems connected with health, but information campaigns have not yet shown results of alcohol-related problems at thermal spas. The aim of this study was to assess the mean daily alcohol intake (grams/day) in a thermal spa population. The sample was conducted over a 6 month period and consisted of 290 subjects. The mean age was 59.01 years (range 23-89, SD +/-14.20) (150 males and 140 females). All of the subjects underwent a medical examination free of charge and an alcohol case history report. The medical staff working at the spa included two doctors, a biologist, a cardiologist, and a dietician. In the final 176 subjects studied, the mean daily alcohol intake was 32.33 g/day, significantly higher in the males than the females (p < 0.001). The 176 subjects were subsequently divided into three subgroups according to their daily alcohol intake: group A, <40 g/day; group B, 40 to 79 g/day; group C, > or =80 g/day. The number of subjects in group A was significantly greater than in group B and group C (p < 0.001). It is important to emphasize that 33.5% of the subjects studied presented a mean daily alcohol intake of > or =40 g/day and therefore were more exposed to the risk of the onset of alcohol-related problems. In view of these results, thermal spas should thus be considered an additional place in which to identify subjects at risk for problematic use of alcohol and represent an environment in which primary prevention campaigns could be started. PMID- 9726277 TI - Sustained-release naltrexone for alcoholism treatment: a preliminary study. AB - This 12-week study examined the bioavailability, tolerability, and potential efficacy of an injectable sustained-release preparation (SRP) of naltrexone (NTX). Twenty alcohol-dependent subjects took NTX 50 mg po daily for 2 weeks, followed by a 2-week, no-medication Washout Period, a 4-week Injection Period, and a 4-week Follow-up Period. Fifteen subjects (75%) received a single subcutaneous injection of 206 mg of sustained-release NTX, and five subjects (25%) received a placebo injection. All subjects also received eight weekly coping skills sessions during the Oral NTX, and the Washout and Injection Periods. RESULTS: After injection, NTX plasma concentrations exceeded a mean of 1 ng/ml for 21 days. Adverse effects produced by the SRP of NTX were comparable with those resulting from oral NTX therapy. Compared with placebo, the SRP of NTX significantly reduced the frequency of heavy drinking days during the Injection and Follow-up Periods. CONCLUSIONS: The results of this preliminary study support the potential clinical utility of the SRP of NTX for treatment of alcohol dependence. PMID- 9726278 TI - A family-based analysis of whether the functional promoter alleles of the serotonin transporter gene HTT affect the risk for alcohol dependence. AB - A population association between a regulatory variation in the promoter of the serotonin transporter gene (HTT) and severe alcohol dependence was recently reported. We analyzed this potential association in a large number of systematically ascertained families in the United States; these families had at least three first-degree relatives who were alcohol-dependent. Analyses focused on individuals defined as alcohol-dependent by criteria from ICD-10 and on subsets of these individuals reporting withdrawal-related symptoms. Application of the transmission disequilibrium test did not provide support for either linkage or association between this functional polymorphism and alcohol dependence; there was no significant bias in the transmission of either allele to the alcohol-dependent offspring. We also report that African Americans differ from Caucasians in allele frequencies for this polymorphism. PMID- 9726280 TI - Intravenous self-administration of ethanol in beta-endorphin-deficient mice. AB - Extensive research on both human alcoholics and in animal models of alcoholism has implicated the release of endogenous opioids in the consumption of ethanol. Various experiments using opioid antagonists have indicated that these drugs cause both humans and animals to decrease their consumption of ethanol. However, it remains unclear exactly which of the endogenous opioids mediates the rewarding effects of ethanol. The present experiment used intravenous self-administration of ethanol to determine whether beta-endorphin (BE)-deficient mice differed from wild-type (WT) mice in ethanol self-administration. The BE-deficient mice completely lack BE, but are otherwise similar to the WT mice. By using intravenous self-administration, we were able to rule out any ability of BE to mediate differences in ethanol consumption via palatability factors alone. Both types of mice were 7 generations backcrossed onto a C57BL/6J inbred strain background. During nine daily, 2-hr free-operant sessions, 14 BE-deficient and 17 WT mice could nosepoke for 75 mg/kg ethanol infusions delivered intravenously on an fixed-ratio 3 schedule with a 2-sec time-out after each reinforcer delivery. Reinforcer delivery occurred following nosepokes in only one of two holes. Contrary to what was expected, BE-deficient mice acquired selective operant responding for ethanol, whereas WT mice did not. Although the two genotypes did not differ in either operant or locomotor behavior during the first session, by the end of the nine sessions, BE deficient mice were reliably nosepoking for ethanol, whereas WT mice were not. These findings may indicate that BE is not essential for the postingestive reinforcing effects of ethanol in these animals. PMID- 9726279 TI - Ethanol acutely decreases calcium transients in cultured human myotubes. AB - Ethanol consumption frequently leads to a number of skeletal muscle disorders, including acute and chronic alcoholic myopathy. Ethanol has been found to interfere with signal transduction mechanisms in cardiac and smooth muscle cells. We studied the effects of ethanol on the intracellular calcium ([Ca2+]i) transients responsible for excitation-contraction coupling in human myotubes from chronic alcoholic patients and healthy controls. Cultured myotubes were loaded with the fluorescent Ca2+ indicator fura-2 and evaluated on a single-cell basis. Following electrical stimulation, ethanol caused a significant reversible dose dependent reduction in [Ca2+]i transient amplitude, achieving a mean decrease of 36+/-5% at 300 mM ethanol (p < 0.01), without modifying the basal [Ca2+]i. This acute effect of ethanol was similar in myotubes obtained from chronic alcoholics and controls. Similarly, ethanol caused a dose-dependent reduction of [Ca2+]i transient amplitude in control samples when depolarization was elicited by 100 mM KCl (p < 0.01). Several potential mechanisms of ethanol action were studied in control muscle samples. Sarcolemmal Ca2+ entry was measured indirectly by monitoring Mn2+-quenching of intracellular fura-2 via the nitrendipine-sensitive Ca2+ channels during electrical pacing. Ethanol at doses of 100 mM and greater caused a dose-dependent reduction in the rate of quench (p < 0.01). In addition, the intracellular pool of Ca2+ releasable by caffeine was found to be reduced at 300 mM ethanol (p < 0.05). We conclude that ethanol reduces the [Ca2+]i transients underlying excitation-contraction coupling in human myotubes, and that this occurs to a similar extent in cells obtained from chronic alcoholics and controls. This acute effect of ethanol was primarily due to an inhibitory effect of ethanol on sarcolemmal Ca2+ influx via voltage-operated Ca2+ channels, although there may also be an effect on the Ca2+ sarcoplasmic reticulum loading state. PMID- 9726281 TI - Confirmation of quantitative trait loci for alcohol preference in mice. AB - An F2 intercross derived from C57BL/6 and DBA/2 progenitor inbred strains was used to test for replication of quantitative trait loci (QTLs) for alcohol preference nominated by a previous study using BXD recombinant inbred (RI) strains (Rodriguez et al., Alcohol. Clin. Exp. Res. 19:367-379, 1995). Fourteen provisional QTLs were nominated in the original RI study with a p < 0.05 criterion. In the present study, a genome scan (101 microsatellite markers) was conducted on an F2 population (n = 218). Three significant QTLs were detected on chromosomes 1, 4, and 9, and three suggestive QTLs were detected on chromosomes 2, 3, and 10. Of these six QTLs, four were consistent with the previous RI nominations. The replication rate of 28.6% (4 of 14) is in agreement with the results of simulation studies performed by Belknap et al. (Behav. Genet. 26:149 160, 1996) and supports the methodological argument for a multistage research design for nominating and replicating QTLs. PMID- 9726282 TI - The reinforcing effects of ethanol are altered by the endogenous neurosteroid, allopregnanolone. AB - We examined the effect of systemic administration of the endogenously occurring progesterone metabolite, allopregnanolone, on oral self-administration of ethanol by male rats. Rats were trained to perform an operant response for presentation of 0.1 ml of a solution of 10% ethanol in water using the sucrose fading technique. After acquisition of stable lever-press responding on a fixed-ratio 4 schedule, subjects received subcutaneous injections of 1, 3, or 10 mg/kg of allopregnanolone, or vehicle, 20 min prior to the self-administration session. Pretreatment with 3 mg/kg, but not 1 or 10 mg/kg, increased the mean total number of lever press responses made to obtain ethanol, and therefore increased the mean total number of ethanol presentations. The number of responses and response rate were examined as a function of the number of "runs" within the 30-min session; a "run" was defined as a series of consecutive responses with an interresponse interval of <1 min. The increase in total responses after 3 mg/kg was due in part to an increased number of responses for the first run of the session, with no effect on response rates. However, the higher dose of 10 mg/kg decreased response rates within the first run. Thus, allopregnanolone alters ethanol-reinforced responding at concentrations lower than those that depress rates of responding. The effects of administration of the benzodiazepene, diazepam, were determined for comparison with those of the neurosteroid. The subcutaneous injection of 0.3, 1.0, or 3.0 mg/kg of diazepam did not produce any clear dose-dependent changes in measures of ethanol-reinforced operant responding, supporting the suggestion of differences in the contribution of the benzodiazepene and neurosteroid binding sites to GABA(A) receptor function. The results indicate that exogenous administration of allopregnanolone dose-dependently alters ethanol-reinforced operant responding, and suggest that this endogenously occurring neurosteroid could mediate some of the reinforcing effects of ethanol. PMID- 9726283 TI - Role of catalase in in vitro acetaldehyde formation by human colonic contents. AB - Ingested ethanol is transported to the colon via blood circulation, and intracolonic ethanol levels are equal to those of the blood ethanol levels. In the large intestine, ethanol is oxidized by colonic bacteria, and this can lead to extraordinarily high acetaldehyde levels that might be responsible, in part, for ethanol-associated carcinogenicity and gastrointestinal symptoms. It is believed that bacterial acetaldehyde formation is mediated via microbial alcohol dehydrogenases (ADHs). However, almost all cytochrome-containing aerobic and facultative anaerobic bacteria possess catalase activity, and catalase can, in the presence of hydrogen peroxide (H2O2), use several alcohols (e.g., ethanol) as substrates and convert them to their corresponding aldehydes. In this study we demonstrate acetaldehyde production from ethanol in vitro by colonic contents in a reaction catalyzed by both bacterial ADH and catalase. The amount of acetaldehyde produced by the human colonic contents was proportional to the ethanol concentration, the amount of colonic contents, and the length of incubation time, even in the absence of added nicotinamide adenine dinucleotide or H2O2. The catalase inhibitors sodium azide and 3-amino-1,2,4-triazole (3-AT) markedly reduced the amount of acetaldehyde produced from 22 mM ethanol in a concentration dependent manner compared with the control samples (0.1 mM sodium azide to 73% and 10 mM 3-AT to 67% of control). H2O2 generating system [beta-D(+) glucose + glucose oxidase] and nicotinamide adenine dinucleotide induced acetaldehyde formation up to 6- and 5-fold, respectively, and together these increased acetaldehyde formation up to 11-fold. The mean supernatant catalase activity was 0.53+/-0.1 micromol/min/mg protein after the addition of 10 mM H2O2, and there was a significant (p < 0.05) correlation between catalase activity and acetaldehyde production after the addition of the hydrogen peroxide generating system. Our results demonstrate that colonic contents possess catalase activity, which probably is of bacterial origin, and indicate that in addition to ADH, part of the acetaldehyde produced in the large intestine during ethanol metabolism can be catalase dependent. PMID- 9726284 TI - In vivo induction of tyrosylprotein sulfotransferase by ethanol: role of increased enzyme synthesis. AB - Tyrosine sulfation is a posttranslational modification involved in the synthesis, secretion, and biological activity of proteins and peptides. Our previous studies have demonstrated that the enzyme activity was induced by ethanol. In the present work, the induction was studied in detail. Initial experiments were conducted to examine the time course of tyrosylprotein sulfotransferase (TPST) induction in rats pair-fed liquid diets containing either ethanol or carbohydrate substitute (controls). Marked elevation of TPST activity (3-fold) was measured on day 10 in the liver and gastric mucosa of ethanol-fed rats. Ethanol-mediated enhancement was also noticed by Western-blot analysis with anti-TPST antibody in both the liver and gastric mucosa on days 5 and 10. We then determined the steady-state TPST protein turnover in ethanol-fed and control animals that were given 35S methionine after 10 days of pair-feeding with liquid diet. The rates of TPST synthesis assessed by measuring initial rates of incorporation of 35S-methionine into TPST was increased in the liver and gastric mucosa of animals fed with ethanol. Monophasic exponential decay curves showed that TPST protein half-lives for liver (control: 34 hr, ethanol: 32 hr) and gastric mucosa (control: 52 hr, ethanol: 48 hr) did not differ between control and ethanol groups. Our overall results indicate that the in vivo induction of TPST by ethanol involves increased enzyme synthesis rather than decreased enzyme degradation. PMID- 9726285 TI - Quantitation of the mass of fatty acid ethyl esters synthesized by Hep G2 cells incubated with ethanol. AB - Fatty acid ethyl esters (FAEE), esterification products of fatty acids and ethanol, have been increasingly implicated as mediators of ethanol-induced organ damage. The first goal of this study was to determine the mass of FAEE synthesized by Hep G2 cells exposed to a given dose of ethanol. The second goal was to determine whether all fatty acids in cells are equally available for FAEE synthesis. Hep G2 cells and essential fatty acid deficient Hep G2 cells (Hep G2 EFD) were used to study the synthesis of FAEE upon exposure to ethanol. A two pool fatty acid model was created: (1) a "previously incorporated pool" formed by incubating the cells with 14C-labeled fatty acids for 24 hr; and (2) a "newly incorporated pool" formed by incubating cells with 3H-labeled fatty acids for 0.5 hr. The FAEE production from each pool was then determined. The total production of FAEE within 3 hr by Hep G2 cells in culture was 150 to 250 pmol/mg cell protein. The fatty acids most recently incorporated into the cells were preferred as substrates for FAEE synthesis because a higher percentage of fatty acids from the newly incorporated pool was used for FAEE synthesis than from the previously incorporated pool. Furthermore, a dose-response relationship was observed between the amount of fatty acid in the newly incorporated pool and FAEE production, but not between the amount of fatty acid in the previously incorporated pool and FAEE synthesis. Taken together, the results indicate that a relatively small amount of endogenously synthesized FAEE is generated from specific intracellular pools of fatty acid since not all fatty acids are equally available for FAEE synthesis. This indicates that if endogenous FAEE are toxic, they exert their toxic effect at very low intracellular FAEE concentrations. PMID- 9726286 TI - Effects of ethanol on recombinant glycine receptors expressed in mammalian cell lines. AB - We examined the effects of acute ethanol exposure on recombinant human glycine receptors transiently transfected into HEK 293 cells and stably transfected into Ltk- fibroblast-like cells. In our study of the effects of ethanol, we used the whole-cell patch-clamp configuration. Relatively low concentrations of ethanol (25 mM and 50 mM) did not affect glycine-gated currents in any of the cell lines studied. Higher concentrations of ethanol (100 mM and 200 mM) significantly potentiated glycine responses only in stably transfected Ltk- cells expressing alpha1 and alpha2 subunits and in HEK 293 cells transiently expressing alpha2 subunits. Cells stably expressing alpha1 versus alpha2 glycine receptors were modulated equally by ethanol. Both glycine alpha1 and glycine alpha1beta receptors transiently expressed in HEK 293 cells were insensitive to all concentrations of ethanol tested; however, there was a trend toward potentiation at 100 and 200 mM ethanol concentrations. A population of cells (41-87%) that was sensitive to the potentiating effects of 100 and 200 mM ethanol (defined as more than 10% potentiation) was identified in both cell lines tested. In these sensitive cells, ethanol (100 and 200 mM) produced significant potentiation, independent of the cell line and the glycine receptor subunit tested. Together with published results from studies with Xenopus oocytes, these data indicate that the sensitivity of recombinant glycine receptors to ethanol depends upon the expression system. PMID- 9726288 TI - Taste reactivity to alcohol and basic tastes in outbred mice. AB - The taste reactivity test was used to determine the response of outbred mice to orally infused taste solutions. For the initial measures, mice (n = 10) were tested with 3%, 6%, 9%, and 12% (v/v) alcohol and four taste solutions: sucrose, sodium chloride, hydrochloric acid, and quinine hydrochloride (a single concentration of each). A second group of naive mice (n = 16) was tested with 5%, 10%, 20%, 30%, and 40% alcohol. The final set of measures with naive mice (n = 26) was taken with a range of sucrose concentrations: 0.01 M, 0.05 M, 0.1 M, 0.5 M, and 1.0 M. In general, mice made similar reactivity responses to all solutions tested. A predominant component of the mouse response to all infused fluids was forelimb flailing; gaping was also a common response to all solutions. Despite the large number of aversive-type responses, mice rejected very little fluid via passive drip or fluid expulsion. The single, significant difference in responding to the four taste stimuli was that mice made fewer aversive responses to sucrose. Differential responding to the 5 to 40% alcohol concentrations and sucrose concentrations was observed. Mice increased ingestive responding as the concentration of alcohol and sucrose increased. Aversive responding decreased reliably only with increases in the sucrose concentration. Data provide the first reported taste reactivity responses of mice to orally infused taste solutions. These results can be compared with the extant data available in rats and can also be used as a basis for exploring taste factors in genetically defined mouse populations. PMID- 9726287 TI - Electrophysiological characterization of cerebellar neurons from adult rats exposed to ethanol during development. AB - The purpose of this study was to investigate the spontaneous activity of mature rat cerebellar neurons that had been exposed to ethanol (EtOH) during postnatal days 4 to 10, which corresponds to the third trimester in humans. Newborn Sprague Dawley rats were implanted with gastric feeding tubes and were artificially reared from postnatal days 4 to 10 with two different diets. The experimental group received 4.5 g/kg/day of EtOH delivered in a milk solution. Controls received similar feeding with an isocaloric supplement replacing the EtOH. Electrophysiological evaluations were performed after an EtOH-free rearing period. Although lobules IX and X of the cerebellar vermis appeared morphologically smaller in the animals neonatally exposed to EtOH, compared with controls, extracellular recordings from both Purkinje cells and Golgi interneurons in adult rats showed no differences in spontaneous activity or firing pattern between the control and EtOH-exposed animals. Similarly, excitations and inhibitions of Purkinje neuron activity evoked by parallel pathway stimulation appeared unaffected by the developmental EtOH exposure. However, we did observe a significant decrease in the proportion of Purkinje neurons generating complex spike bursts in the group exposed to EtOH neonatally. These data suggest that, although fewer Purkinje neurons may survive the brain growth spurt if exposed to EtOH during this critical period of development, those that do survive appear to function normally. The observed abnormality in complex spike production may result from EtOH effects on developing neurons in the inferior olive that give rise to the climbing fibers that cause this bursting pattern in Purkinje neurons. PMID- 9726289 TI - Effects of chronic ethanol consumption and aging on proenkephalin and neurotensin. AB - We examined the combined effects of chronic ethanol consumption and aging on mRNA and peptide for proenkephalin (PE), the precursor of met- and leu-enkephalin. This study also evaluated the effects of aging and alcohol on the level of the mRNA encoding the common precursor of neurotensin (NT) and neuromedin N (NN). PE mRNA and NT/NN mRNA were quantitated in multiple brain areas of 5- and 24-month old male Fischer 344 rats. Aging, but not chronic ethanol consumption, altered PE mRNA and peptide levels. Aging was accompanied by a loss of PE peptide and PE mRNA in the rostral striatum. In aged rats, PE mRNA was also reduced in the shell region of the nucleus accumbens. The decline in PE mRNA in the rostral striatum and shell region of the nucleus accumbens was caused by a reduction in the number of cells that contain PE mRNA. The percentage of PE mRNA-containing neurons that express a high amount of PE mRNA was also lower in the rostral striatum of 24 month-old rats. The effects of aging may impair motor function and alter the rewarding properties of ethanol consumption. Neither aging nor alcohol changed the PE mRNA levels in the core region of the nucleus accumbens, the frontal cortex, and the piriform cortex. In contrast to PE mRNA, neither aging nor chronic ethanol consumption affected NT/NN mRNA in the regions analyzed. Normal NT/NN mRNA levels were found in the lateral septum and two hippocampal brain areas: the dorsal subiculum and CA1 regions. PMID- 9726290 TI - Inhibition of intracolonic acetaldehyde production and alcoholic fermentation in rats by ciprofloxacin. AB - Heavy drinking is associated with many gastrointestinal symptoms and diseases, such as rapid intestinal transit time, diarrhea, colon polyps, and colorectal cancer. Acetaldehyde produced from ethanol by intestinal microbes has recently been suggested to be one of the pathogenetic factors related to alcohol associated gastrointestinal morbidity. Furthermore, acetaldehyde is absorbed from the colon into portal blood and may thus contribute to the development of alcoholic liver injury. The present study was aimed to investigate the significance of gut aerobic flora in intracolonic acetaldehyde formation. For this study, 58 male Wistar rats (aged 9 to 11 weeks) were used. Half of the rats received ciprofloxacin for four consecutive days. Control rats (n = 29) received standard chow. On the fifth day of treatment, 1.5 g/kg body weight of ethanol was administered intraperitoneally to 19 rats receiving ciprofloxacin and 19 control rats. Ten ciprofloxacin-treated and 10 control rats received equal volumes of physiological saline intraperitoneally. Two hours after the injection of ethanol or saline, the samples of colonic contents and blood were obtained. Acetaldehyde and ethanol levels of the samples were determined by headspace gas chromatography. The intracolonic acetaldehyde level 2 hr after ethanol administration was 483+/-169 microM (maximum: 2.7 mM). High intracolonic acetaldehyde after ethanol injection was significantly reduced by ciprofloxacin treatment. After ciprofloxacin, intracolonic acetaldehyde levels before and after the injection of ethanol were 25+/-4.8 and 23+/-15 microM, respectively. Ciprofloxacin treatment resulted also in significantly higher blood (p < 0.005) and intracolonic (p < 0.0001) ethanol levels than in the control animals. Furthermore, ciprofloxacin treatment totally abolished the formation of endogenous ethanol in the large intestine. This study demonstrates that alcoholic fermentation and intracoIonic acetaldehyde production can be blocked by diminishing the amount of intracolonic aerobic bacteria with ciprofloxacin. Our findings indicate that the bacteriocolonic pathway for ethanol oxidation is mediated almost exclusively by gut aerobic microbes, and this knowledge may provide new insights into the studies on the pathogenesis of alcohol-related gastrointestinal symptoms and diseases. PMID- 9726291 TI - Microsomal acetaldehyde oxidation is negligible in the presence of ethanol. AB - The microsomal ethanol oxidizing system (MEOS), inducible by ethanol and acetone, oxidizes ethanol to acetaldehyde, which causes many toxic effects associated with excess ethanol. Recent studies reported that rat liver microsomes also oxidize acetaldehyde, thereby challenging the validity of the assessment of MEOS activity by measuring acetaldehyde production and suggesting that MEOS activity results in the accumulation not of acetaldehyde but, rather, of its less toxic metabolite, acetate. To address these issues, we compared both metabolic rates of ethanol and acetaldehyde and the effect of ethanol on the acetaldehyde metabolism. Liver microsomes were prepared from Sprague-Dawley rats induced either with acetone for 3 days or ethanol for 3 weeks. NADPH-dependent acetaldehyde (300 microM) metabolism was measured in two ways: (1) by detection of acetaldehyde disappearance by headspace gas chromatography, and (2) by assessment of acetaldehyde oxidation by liquid scintillation counting of acetate formed from [1,2-14C]acetaldehyde. Ethanol (50 mM) oxidation was measured by gas chromatography. In acetone- and ethanol-induced rat liver microsomes, the acetaldehyde disappearance (p < 0.0001) and oxidation (p < 0.0001) rates were both significantly increased. The rates of acetaldehyde oxidation paralleled those of p-nitrophenol hydroxylation (r = 0.974, p < 0.0001), with a Km of 82+/ 14 microM and a Vmax of 4.8+/-0.5 nmol/min/mg protein in acetone-induced microsomes. Acetaldehyde disappearance in acetone-induced microsomes and acetaldehyde oxidation in acetone-induced and ethanol-induced microsomes were significantly lower than the corresponding ethanol oxidation, with rates (nmol/min/mg protein) of 4.6+/-0.6 versus 9.0+/-0.8 (p < 0.005), 4.4+/-0.3 versus 9.1+/-0.5 (p < 0.0005), and 14.0+/-0.9 versus 19.5+/-1.8 (p < 0.05), respectively. The presence of 50 mM ethanol decreased this metabolism to 0.9+/ 0.3 (p < 0.005), 0.5+/-0.1 (p < 0.001), and 1.8+/-0.3 (p < 0.001), resulting in rates of acetaldehyde metabolism of only 9.8+/-3.2%, 6.0+/-0.5%, and 9.5+/-1.2% (respectively) of those of ethanol oxidation. In conclusion, rat liver microsomes oxidize acetaldehyde at much lower rates than ethanol, and this acetaldehyde metabolism is strikingly inhibited by ethanol. Accordingly, acetaldehyde formation provides an accurate assessment of MEOS activity. Furthermore, because acetaldehyde production vastly exceeds its oxidation, the net result of MEOS activity is the accumulation of this toxic metabolite. PMID- 9726292 TI - The alcohol deprivation effect in the alcohol-preferring P rat under free drinking and operant access conditions. AB - The alcohol-deprivation effect (ADE) was examined under 4-hr operant and 24-hr free-choice alcohol access in the alcohol-preferring (P) rat after deprivation intervals from 2 to 4 weeks. Results indicated that adult male P rats responding for 6 weeks on a concurrent FR-5/FR-1 schedule of reinforcement for alcohol and water, respectively, and then deprived of alcohol for 2 weeks, demonstrated a 40% increase in alcohol responding during the first 60 min of alcohol reinstatement. The alcohol deprivation effect was temporary, however, as responding did not differ from baseline levels on the second day of reinstatement. In a second experiment, weanling male and female P rats received 7 weeks of continuous access to alcohol, beginning at 21 days of age, and were then deprived of alcohol for 4 weeks. On the first day of alcohol reinstatement, P rats exhibited a 40% to 45% increase from baseline alcohol drinking levels, with alcohol intake returning to baseline levels by the 3rd day of reinstatement. Although alcohol intake was higher in females than in males when adjustment was made for body weight, there were no gender differences in the magnitude of the alcohol deprivation effect. Taken together, these results indicate that the ADE is a long-lasting phenomenon that occurs under both operant and continuous access conditions in the P rat, and thus these rats may be useful models for the study of factors involved in relapse of alcohol drinking. PMID- 9726293 TI - Defective intracellular processing of lactase-phlorizin hydrolase protein in rats prenatally exposed to ethanol. AB - We have previously shown that fetal exposure to ethanol in rats produces both structural and biochemical abnormalities in absorptive enterocytes. Among the indicators of injury are derangements in the expression of lactase-phlorizin hydrolase (LPH), which is an essential enzyme for the assimilation of milk. In an animal model of fetal alcohol syndrome, unsuckled newborn rats prenatally exposed to maternal ethanol revealed a 10- to 15-fold increase in the number of LPH mRNA molecules per absorptive enterocyte, compared with controls (Estrada et al., Alcohol. Clin. Exp. Res. 20:1662-1668, 1996). However, lactase activity per cell was similar in both groups. The aim of this study was to characterize the effect of prenatal exposure to ethanol on the processing of LPH mRNA and protein. RNase protection assays using 3'- and 5'-directed antisense RNA probes revealed that the LPH mRNA from ethanol-exposed pups is full length. However, metabolic labeling, followed by immunoprecipitation using an anti-LPH monoclonal antibody, demonstrated a significant alteration in LPH protein processing. Intestinal explants from 21-day ethanol-exposed fetuses that were chased 30 min after a [35S]methionine pulse showed greater amounts of newly synthesized LPH precursors (205 and 220 kDa) and low molecular weight degradation products than controls. However, despite the increases in LPH precursor, the amount of 130 kDa mature LPH was similar in ethanol-exposed and control explants. These data suggest an increase in intracellular degradation of LPH precursor in rats prenatally exposed to ethanol, which occurs before its insertion into the microvillus membrane. Biosynthesis of LPH appears to be upregulated at the transcriptional level, which overcomes the degradation of LPH precursor during processing. PMID- 9726294 TI - Results of elective lumbar discectomy for patients involved in the workers' compensation system. AB - We compared the outcomes from lumbar discectomy for patients who were workers' compensation claimants and/or who were involved in active litigation with patients who underwent elective lumbar discectomy, but who were not involved with either compensation or litigation. Eighty-two consecutive patients who underwent elective lumbar discectomy by the senior author were identified from 1989 through 1994. Those patients who underwent a primary discectomy with a minimum of 6 months' follow-up were studied. Patients were excluded if a spinal fusion was performed or if a multilevel laminectomy procedure was required. Patients were classified as compensation patients if they were involved in either worker's compensation claims or active litigation at the time of the lumbar discectomy. The compensation group was further divided into three subsets of patients: those involved in active litigation without compensation, those involved in both compensation and litigation, and those pursuing workers' compensation claims without litigation. The control group was comprised of patients who were not in any way involved with compensation or litigation. Outcome assessment and ratings were determined independently by the coauthors, not the primary surgeon. Outcome was based on pain, employment status, analgesic use, and level of activity. Fifty four patients met the inclusion criteria. Average follow-up for the compensation patients was 40 weeks. Follow-up for the noncompensation patients averaged 51 weeks. Eighty-one percent of our patients in the noncompensation group achieved a good result. Only 1 of 27 patients was categorized as having a poor outcome. Conversely, patients who were actively involved in the compensation and/or litigation process had significantly poorer outcomes, with only 29% of the patients receiving a good outcome evaluation (p = < 0.0002). Legal involvement was associated with poorer outcome in compensation patients (p = < 0.001). PMID- 9726295 TI - Motor deficit in lumbar spinal stenosis: a retrospective study of a series of 50 patients. AB - Severe motor weakness is an infrequent symptom in the course of lumbar stenosis. The objectives of this study are threefold: to describe the motor deficit, evaluate the prognosis factors, and determine the type of stenosis most likely to be complicated by motor loss. Fifty consecutive patients with a mean age of 65 years, operated on for a lumbar stenosis and with a severe motor deficit, have been retrospectively studied with a mean follow-up of 38 months. The overall functional result was evaluated according to the Beaujon scoring system. The motor capacity was rated from 0 (complete paralysis) to 5 (normal strength). Prognosis factors were investigated with a multivariate analysis model. Motor weakness was rated as zero 11 times, as one 8 times, as two 8 times, and as three 23 times. According to our rating scale, the overall results were considered excellent in 25 cases, good in 17 cases, and fair in the 8 remaining cases. Regression of motor weakness was complete 15 times, partial 25 times, and null 10 times. In this study, favorable prognosis parameters of motor weakness recovery were as follows: association with a discal herniation, stenosis at one level, preoperative duration of motor weakness <6 weeks, age <65, and monoradicular deficit. In contrast, severity of the initial motor weakness, association with sphincter abnormalities, presence or not of degenerative spondylolisthesis, or of a complete block on the myelogram were not influential variables. PMID- 9726296 TI - Usefulness of MRI in isolated upper cervical spine fractures in adults. AB - A prospective analysis of patients admitted with isolated upper cervical spine fractures and who had magnetic resonance (MR) imaging performed within 48 h of the inciting traumatic event was completed to determine the clinical usefulness and cost effectiveness of routine MR screening. In patients with an identified neurologic deficit, MR findings changed the treatment of 25% (one of four) of the patients, whereas MR findings did not change the treatment of any patient identified without a neurologic deficit. We recommend that in adult patients with an isolated upper cervical spine fracture, MR should not be routinely ordered in patients without a neurologic deficit. This advanced imaging modality is not a useful or cost-effective screening device for patients presenting with a fracture of the upper cervical spine without neurologic deficit. PMID- 9726298 TI - Augmentation of anterior transvertebral screws using threaded teflon anchoring. AB - This is a biomechanical study to investigate the effect of augmentation of single anterior transvertebral screws. Two subsequent experiments (pullout and caudal loading) were performed in 78 porcine vertebral bodies of equal bone mineral density using 6.5-mm transvertebral screws augmented or not with specially designed Teflon anchoring. Three different types of 35- and 45-mm unaugmented screws (Kaneda, TSRH, and Zielke) were inserted at 90 degrees laterally in the vertebral body and were tested for pullout force. The pullout load to loosen the three different unaugmented screws did not significantly differ. The pullout force to loosen the 35- and 45-mm Zielke screws was 507 +/- 22 and 860 +/- 50 N, respectively. The corresponding pullout load to loosen the construct Zielke screw Teflon anchoring was 1,005 +/- 148 N and 1,306 +/- 135 N for the 35- and 45-mm screws, respectively. When comparing the pullout force needed for unaugmented versus the augmented Zielke screw of the same length, there was a statistically significant (p < 0.0001) difference. During the caudal loading test, the unaugmented 45-mm Zielke screw showed that a uniform slope up to the yield point occurred at 0.35 +/- 0.12 mm of displacement, with an average tilting force for the tested screw of 1,362 +/- 151 N, when the screw became loose. The caudal loading test for the augmented Zielke screw was interrupted at 2,000 N because the load applied exceeded the load capacity of the testing machine, and thus no loosening occurred. The findings of this in vitro study showed that the Teflon anchoring provides superior anchorage and stability of single transvertebral Zielke screws. However, further biomechanical and clinical studies are required before using this device or its modification. PMID- 9726297 TI - Oblique MRI as a useful adjunct in evaluation of cervical foraminal impingement. AB - Magnetic resonance (MR) imaging is considered the gold standard for soft-tissue disease. The traditional MR imaging series uses axial and sagittal views. The purpose of this study was to demonstrate that oblique MR imaging provides valuable information about the cervical foramen not available from the conventional MR imaging technique. Ten asymptomatic individuals volunteered for MR imaging. Measurements were taken of height, width, and area for the nerves and foramen at the entrance and mid zones. Nerves were graded as normal, contacted, or deformed. Normal foraminal morphology in asymptomatic individuals and characteristics that compromise the space available for the nerve root were identified. Nerves with minimal or no contact had significantly greater foraminal widths than nerves with significant contact. Although there was a correlation between nerve contact and foraminal width, regression analysis did not demonstrate a correlation between disc height and foraminal size. PMID- 9726299 TI - Foraminal pressure changes during intervertebral distraction simulating anterior cervical discectomy. AB - Distraction of the disc space over baseline height has been shown to increase foraminal size. The purpose of this procedure is to determine pressure changes, with disc space distraction simulating an anterior cervical discectomy and fusion (ACDF). An analysis of pressure changes during disc space distraction at C5-C6 was performed. Data were analyzed for maximal pressure observed and for pressure change with prolonged distraction. Five cadaveric specimens underwent a discectomy at the C5-C6 level. Distraction of the disc space was performed and pressure measurements were obtained from within the foramen. Measurements were made for maximal pressure with an intact posterior longitudinal ligament (PLL), divided PLL, and with the nerve root removed from within the foramen. Pressures were also recorded with prolonged distraction until a steady state was achieved. Incremental distraction of +2, +4, and +6 mm resulted in pressure increases within the foramen. Sectioning of the PLL did not affect these increases. Removal of the nerve root from the foramen resulted in pressure increases; however, these were not significantly different from baseline. Prolonged distraction produced an initial increase and a gradual return toward baseline. Final pressures still differed significantly from baseline. Increase intraforaminal pressures can be seen with increasing disc space distraction such as occurs during an ACDF. This suggests that either the foramen narrows in at least one dimension and/or soft tissue attachments to the nerve produce a tensile force in the nerve as they tighten. The pressure increases relax over time. PMID- 9726300 TI - Torsional injury resulting in disc degeneration: I. An in vivo rabbit model. AB - Torsional injuries may be a precursor to intervertebral disc degeneration, but published rabbit models indicate a latent time of 6 months. We describe a rabbit model in which instability and disc degeneration appear within 3 months. Sixty five male New Zealand rabbits underwent presurgical irradiation to inhibit heterotopic bone formation. Control animals then underwent either a soft-tissue release or facetectomy and capsulotomy, whereas experimental animals received surgery and an acute 30 degrees torsional lumbar injury. Capsulotomy, as well as facetectomy without torsion, failed to effect disc degeneration. However, the rabbits that received torsion exhibited clear indications of degenerative disc changes (thinning, increased PLA2 levels, and decreased nucleus pulposus volume) within 60-90 days. The observations associate disc degeneration with a destabilizing acute torsional injury. PMID- 9726301 TI - Torsional injury resulting in disc degeneration in the rabbit: II. Associative changes in dorsal root ganglion and spinal cord neurotransmitter production. AB - The mechanism mediating the chronic pain associated with lumbar disc degeneration may involve neurotransmitters elaborated by dorsal root ganglion (DRG). This hypothesis has been tested in an applicable rabbit model of disc degeneration. Twenty control male rabbits underwent a soft-tissue release; 20 experimental rabbits sustained a facetectomy and capsulotomy and received an acute torsional lumbar injury. The levels of calcitonin gene-related peptide, vasoactive intestinal peptide, and substance P were measured in the DRG, spinal cord, and disc at 10, 30, 60, and 90 days postoperatively. Torsional injury was associated with a statistically significant increase in most DRG and spinal cord neurotransmitter values after 60-90 days. These points in time marked the periods of maximum biomechanical instability and disc narrowing. Such data support concepts about the association between chronic lumbar spinal instability, disc degeneration, and pain. PMID- 9726302 TI - Pressure measurements in herniated lumbar disks: association with radiologic and clinical data. AB - This prospective observational study of 85 herniated disks was conducted to investigate the relationship between diskomanometry and radiologic and clinical parameters. The mean injection pressure (P0) was 282 kPa, the residual pressure after 60 s (P60) was 181 kPa, and the loss of pressure (LOP) was 34%. P0 and P60 were moderately intercorrelated (r = 0.51). The following associations were significant: low P0 and P60 in advanced annular degeneration and disruption; low P60 and great LOP in large hernias and narrow disks. P0 and P60 were not associated with the pain response at diskography, magnetic resonance signal intensity, or patient age, sex, weight, body mass index, type of occupation, or duration of symptoms. Diskomanometric data seem to be influenced by the level of annular degeneration and disruption, size of herniation and height of the disk space. PMID- 9726303 TI - Biomechanical evaluation of posterior and anterior lumbar interbody fusion techniques. AB - This study determined the biomechanical differences between anterior and posterior lumbar interbody fusion (ALIF and PLIF). Ten cadaveric spines were tested. Five specimens had ALIF and five had PLIF at L4-L5. Stabilization was performed with pedicle screws and rods (Cotrel-Dubboset, Sofamor-Danek, Memphis, TN, U.S.A.). Angular motion was measured in flexion, extension, bending, and torsion on the intact, instrumented, and "fused" specimens. Instrumentation alone caused a significant decrease in segmental motion in all loading modes (p < 0.01). After the simulated fusion procedures, all specimens were most stable in flexion, and significantly less stable in extension (p = 0.04). Comparing directly, ALIF was significant more stable in left torsion (p = 0.03) with trends in left bending (p = 0.08) and right torsion (p = 0.07). Thus, from a purely biomechanical perspective, ALIF appears to be slightly superior to PLIF. PMID- 9726304 TI - Quantitative discomanometry: technique and reproducibility in vitro. AB - Quantitative discomanometry is a study of intradiscal pressure and volume measurements during the injection of fluid into an endplate-disc-endplate complex. The purpose of this article was to describe the technique of quantitative discomanometry, determine the reproducibility of the injection technique in a cadaveric thoracolumbar spine, and standardize the technique for future clinical investigations. Nineteen fresh human cadaveric thoracolumbar discs were injected using quantitative discomanometry to determine: (a) the time necessary for a disc to return to a baseline pressure-volume curve, (b) the reproducibility of the technique in vitro, (c) effects of the injection approach and position of the needle in the disc, and (d) effects of the type and length of tubing as well as gauge of spinal needle. A pressure-volume curve was obtained for each disc injection. Reproducibility was measured by nine parameters obtained from each pressure-volume curve: intrinsic pressure, leakage pressure, initial slope, slope between 0-0.1 ml, slope between 1-4 ml, pressure at 2 ml, pressure at 4 ml, maximum pressure, and volume at maximum pressure. The results demonstrated that (a) the injector apparatus was reproducible, (b) the time necessary for a disc to return to a baseline pressure-volume curve was 24 h, (c) the technique using fresh human cadaveric thoracolumbar discs was reproducible, (d) the anterior and posterolateral approaches had similar results if the needle was placed into the center of the nucleus pulposus using radiographic control, and (e) the type and length of tubing, and gauge of needle did not affect the results. PMID- 9726306 TI - Posttraumatic spinal cord lesions without skeletal or discal and ligamentous abnormalities: the role of MR imaging. AB - The thoracic spine is different from other mobile segments of the spine because of the presence of ribs and their articulations. The rib cage makes the thoracic spine much more stable and, during trauma, provides additional strength and energy-absorbing capacity. This leads to the conclusion that severe trauma is required to damage the thoracic spine, and the skeletal injury is usually evident on radiographs. A spontaneous reducible vertebral luxation (dislocation) is not easy to identify, even with magnetic resonance (MR) imaging. Subtle changes in thoracic spine osseous injuries are not seen on radiographs but may be demonstrated on computed tomography (CT) scans. MR imaging can also demonstrate the posterior ligamentous lesions. In this study, we present three cases of thoracic spinal cord changes without spinal fracture and one disk herniation (degenerative chronic disease). These patients had a permanent neurologic deficit (complete paraplegia); plain radiographs and CT scans showed nothing abnormal. MR imaging showed lesions in the thoracic spinal cord and, in one case, a posttraumatic disk herniation. In cases of post-traumatic cord lesions, MR imaging provides diagnostic information that appears to exceed other imaging modalities. The existence of a neurologic deficit indicates MR as the first examination in cases of traumatic spinal lesions. PMID- 9726305 TI - Pain provocation tests for the assessment of sacroiliac joint dysfunction. AB - A double-blind trial was carried out to determine the sensitivity and specificity of three commonly used pain provocation tests for sacroiliac joint dysfunction. The trial involved 40 patients, all of whom reported pain when they were subjected to each of the three tests. Half of the patients (20) had the symptomatic sacroiliac joint injected with 4 ml of 1% lignocaine, whereas the other 20 patients received 4 ml of normal saline to the painful joint. The level of pain produced by each of the three tests was assessed pre- and posttest injection using a visual analogue scale of 0-100. If the pain could be suppressed by 70% with injection of either normal saline or 1% lignocaine into the symptomatic sacroiliac joint under image intensification, the test was considered to be positive for pain arising from the sacroiliac joint. None of the patients receiving normal saline had their pain suppressed to any significant degree, whereas those patients receiving 1% lignocaine had their pain suppressed sufficiently for the three pain provocation tests to have a specificity of 100% for each test and a sensitivity range of 77-87%. This study indicates that the three tests, when used in combination, have a high predictive value for pain arising from the sacroiliac joint. PMID- 9726308 TI - Three cases of spinal cord tumor originating from the second cervical nerve root. AB - Determination of the level of spinal cord tumors that develop around the foramen magnum or high cervical region can be difficult, because the symptoms of such tumors are quite variable. The surgical approach to use in such cases remains controversial. We describe here three cases of spinal cord tumor originating from the second cervical nerve root. Initial symptoms included occipital pain, glove type numbness, and paresthesia of the upper extremities. In all three cases, myelopathic signs preceded radicular signs. It was possible to resect the tumors almost completely by using a posterior approach with microsurgical technique. Several reports have concerned the best surgical approach for high cervical spinal cord tumors. However, by using our method, tumors originating from the C2 root could be exposed without destroying the facet joints, because the nerve root runs dorsal to the lateral facet joints. We therefore recommend resecting these tumors except when they involve the vertebral artery and facet joints. PMID- 9726307 TI - Monoradiculopathy of the fifth lumbar nerve root due to lumbar disc herniation between lumbar one and lumbar two vertebrae. AB - An extremely rare case is reported of a 34-year-old man who had a drop foot due to a herniated disc between the first and second lumbar vertebrae with a monoradiculopathy of the fifth lumbar nerve root. The diagnosis was made on the basis of myelography and magnetic resonance imaging (MRI), which revealed a disc centrolateral herniation at the level between the first and second lumbar vertebrae. The patient underwent anterior discectomy and fusion with the use of iliac bone graft. Because of increasing local kyphosis and associated symptoms, a posterior TSRH instrumentation was added successfully. Postoperatively the patient had alleviation of his symptoms, and at the 6-year follow-up evaluation, he was completely symptomless. The spine surgeon should be aware of the possibility of this rare location of lower lumbar nerve root compression within the dural sac. In such a case, myelography and MRI seemed to be superior to the computed tomography scan. PMID- 9726309 TI - Familial neurilemoma of the spinal cord in a mother and daughter. AB - Neurilemoma of the spinal cord occurred in a mother and daughter. Case 1 was a 75 year-old woman with gait disturbance. Examination revealed weakness of the lower extremities, and magnetic resonance (MR) imaging showed an intradural extramedullary tumor at T12. After laminectomy, the histologic diagnosis was mixed Antoni type A and B neurilemoma. Case 2 was a 48-year-old woman (daughter of case 1). She presented with cervical pain and numbness of both hands. Examination revealed weakened intrinsic muscles of the right hand and paresthesia of the right upper arm. MR imaging showed a giant hourglass-shaped extradural tumor at C2 and C3. The histologic diagnosis was Antoni type A neurilemoma. Only six families with neurilemoma have been reported, including our patients. Gene analysis of such patients may clarify the etiology of neurilemoma. PMID- 9726310 TI - Paraganglioma of the cauda equina: a case presenting features of increased intracranial pressure. AB - Paragangliomas are benign tumors arising from the heterotopic sympathetic ganglion. They occur more often in the carotid body, glomus jugulare, mediastinum, and paraaortic region; central nervous system locations include the petrous ridge, pineal region, and sella turcica. We report the clinical and imaging features of an unusual case of paraganglioma of the cauda equina. Magnetic resonance imaging (MRI) of the thoracolumbar region should be performed in cases of increased intracranial pressure whenever the head examination does not reveal the exact cause of the problem. PMID- 9726311 TI - Hybrid total hip arthroplasty in patients under the age of fifty: a five- to ten year follow-up. AB - In 37 patients, 45 total hip replacements were performed using contemporary cementing techniques, an uncemented Harris-Galante I acetabular component and a cemented precoated Iowa femoral component in patients under the age of 50 at the time of their surgery; 36 patients with 43 hybrid hips were living, 1 patient with 2 hybrid hips was decreased. No patients were lost to follow-up. At 5- to 10 year follow-up, eight hips were revised for aseptic loosening. No acetabular components, and eight femoral components (18%) were revised for aseptic loosening. When looking at radiographic results, including revision as well as those components that were probably or definitely loose on radiographs, 0 acetabular components and 11 femoral components (24%) were radiographically loose. These results demonstrate the excellent durability of the uncemented Harris-Galante acetabular component in the younger patient. However, the Iowa grit-blasted methyl methacrylate precoated femoral component had a magnitude increase in the prevalence of revision for aseptic femoral loosening when compared to the senior author's long-term Charnley results in this age group. The authors attribute the failure to the rough surface finish applied to the femoral component. However, the polymethyl methacrylate proximal precoating and the femoral component design may also contribute to the femoral failures. PMID- 9726312 TI - Polyethylene wear in cases using femoral stems of similar geometry, but different metals, porous layer, and modularity. AB - In this report, 83 total hip arthroplasties in 75 patients with femoral stems of similar geometry but different metals, porous surfaces, and femoral head-neck design were compared at a mean follow-up of 66 months (range, 40-104 months). One type of acetabular component and polyethylene were implanted in all hips. The femoral stem was monoblock in 25 hips, and in 58 it had a modular head-neck piece; 70 stems had chrome-cobalt heads, and 13 heads were titanium. Equally satisfactory clinical results were obtained with either type of femoral implant (i.e., modular and monoblock). The calculated average annual linear polyethylene wear was significantly higher for the titanium stems with a plasma-spray porous surface and chrome-cobalt head on a Morse taper than the chrome-cobalt, beaded, monoblock stems (0.22 mm/year vs 0.07 mm/year, P < .0001). The prevalence of periprosthetic osteolysis was higher for these modular stems ( 15.7% vs 0%), but this difference was not statistically significant (P = .09). Gross corrosion was present on the taper surfaces of an autopsy-retrieved femoral implant with a modular cobalt-chrome head on a titanium stem. Particles of chromium 3 orthophosphate were present at the taper rim and in the periarticular tissues. PMID- 9726313 TI - Intraoperative flexion against gravity as an indication of ultimate range of motion in individual cases after total knee arthroplasty. AB - To assess a method of predicting the final postoperative flexion in individual cases after total knee arthroplasty, 364 primary posterior cruciate-retaining total knee arthroplasties were reviewed retrospectively. The knees were subdivided into three preoperative flexion groups--I: poor motion (0 degrees to 85 degrees), II: intermediate motion (90 degrees to 110 degrees), and III: good motion (115 degrees to 140 degrees). There were 302 cases of osteoarthritis and 62 rheumatoid knees (12 juvenile rheumatoid). Correlation was made between preoperative; intraoperative, and postoperative (minimum 2-year follow-up) passive knee flexion for individuals. Intraoperative flexion against gravity was measured after capsular closure by passively flexing the patient's hip 90 degrees and allowing the weight of the lower leg to flex the knee joint. The overall mean value of postoperative flexion for all three groups was similar to preoperative and intraoperative flexion in both osteoarthritis and rheumatoid arthritis. In the poor motion group (I), postoperative flexion (103 degrees) was increased over preoperative flexion (84 degrees) but similar to intraoperative flexion (104 degrees). In the intermediate group (II), postoperative flexion (110 degrees) was similar to both the preoperative flexion (108 degrees) and intraoperative flexion (110 degrees). In the good group (III), postoperative flexion (119 degrees) tended to be less than preoperative flexion (123 degrees) and more than intraoperative flexion (116 degrees), but the differences were not statistically significant. When comparing preoperative and intraoperative flexion to postoperative flexion for individual cases, 55% of knees had postoperative flexion +/-10 degrees of their preoperative value, while 97% of knees had postoperative flexion +/-10 degrees of their intraoperative value. This study indicates that the final postoperative mean flexion for a group of patients with poor preoperative flexion (<85 degrees) and for individual cases (regardless of their preoperative mobility) can best be predicted by intraoperative flexion against gravity rather than by a preoperative value. PMID- 9726314 TI - Opportunities for control of hospital costs for total joint arthroplasty after initial cost containment. AB - Financial analysis of 391 joint replacement operations performed during 1996 determined that almost 80% of the hospital cost for joint replacement procedures was generated in the operating room, nursing units, recovery room, and pharmacy during the first 48 hours of hospitalization. Attempts to control or reduce the hospital cost of joint replacement operations should focus on these specific areas of opportunity. PMID- 9726315 TI - Examination of wear in Duraloc acetabular components: two- to five-year evaluation of Hylamer and Enduron liners. AB - To help increase the longevity of polyethylene components used in total joint arthroplasty, new forms of ultrahigh molecular weight polyethylene (UHMWPE) have been developed. This study reports on the in vivo performance of Hylamer (DePuy DuPont Orthopaedics, Newark, DE), a high crystalline form of UHMWPE, and on the performance of conventional Enduron polyethylene (DePuy, Warsaw, IN). With the use of a specially designed computer system, the time course of two-dimensional femoral head penetration into 80 Hylamer and 140 Enduron acetabular liners in situ from two to five years was compared. At the most recent follow-up (mean, 3.6 years), the rate of femoral head penetration into the Hylamer liners was lower than the rate of head penetration into the Enduron liners (0.15 mm/year versus 0.20 mm/year, P = . 10). Temporal analysis revealed that the Hylamer liners had lower penetration rates than those of the conventional polyethylene liners in the first three postoperative years, but that head-penetration rates between the two groups became similar with increased time in situ. Continued examination of both types of polyethylene components is imperative before conclusions on long-term performance can be drawn. PMID- 9726316 TI - Universal Total Wrist Implant: experience with a carpal component fixed with three screws. AB - The Universal Wrist Implant was used to treat 31 patients (37 wrists), who had symptoms indicating pancarpal arthritis of the wrist, diagnosed as total wrist arthroplasty. Their mean age was 58.1 years. Follow-up ranged from 48 to 120 months with a mean of 79.4 months (6.7 years). The carpal component of the Universal Total Wrist is fixed to the carpus by titanium screws. Unlike other total wrist prostheses, the primary fixation of the carpal component is in the capitate and not in the third metacarpal. Intercarpal fusion provides a solid bony support for the carpal plate and results in improved longevity. Articular surface of the radial component is inclined 20 degrees, similar to the articular surface of the radius. Components can be inserted with or without cement. In three patients, the prosthesis had to be removed due to infection and persistent dislocation. Of the remaining 34 wrists, 30 (88%) achieved excellent pain relief. Complications occurred in 12 cases (32%). Of these 12 complications 9 (75%) resolved with appropriate treatment. The most common complication with this nonconstrained prosthesis was dislocation. The Universal Total Wrist Implant provides a predictable option to preserve motion and relieve pain when managing wrist joint arthritis. PMID- 9726317 TI - Viability of the acetabular bone bed at revision surgery following cemented primary arthroplasty. AB - Loosening of total hip replacements is often associated with severe loss of periprosthetic bone. The notion exists that the remaining bone is sclerotic, avascular, and displays little osteogenic activity, and that it therefore potentially compromises the revitalization of bone grafts used to restore bony defects. To verify this opinion we studied the bone characteristics in acetabular bone biopsies taken at primary total hip arthroplasty (PTH) and revision total hip arthroplasty (RTH) for a cemented PTH. In 6 PTH patients and in 10 RTH patients, acetabular bone biopsies were taken from the roof, the center, and the lower rim of each acetabulum. Specimens were evaluated by light microscopy and histomorphometrically measured for specimen size, bone area, perimeter, active osteoid perimeter, number of vessels, and osteoclasts. The vascularity and vitality appeared to be comparable in the RTH and PTH bone biopsies. However, the trabecular organization of the RTH bone differed from that of the PTH biopsies. In the PTH biopsies, the trabeculae were running perpendicular to the subchondral bone layer, whereas in the RTH biopsies the layers of bone were oriented parallel to the implant surface. There was abundant remodeling activity in the RTH bone, with large quantities of active osteoid and osteoclasts. These histologic parameters differed, but not statistically significant, from the PTH biopsies. In conclusion, we found that at revision, the acetabular bone was viable with sufficient vascularity and remodeling activity to provide an acceptable recipient host bone bed for revision surgery combined with bone grafting. PMID- 9726318 TI - Effect of acetabular component orientation on recurrent dislocation, pelvic osteolysis, polyethylene wear, and component migration. AB - We retrospectively reviewed 75 total hip arthroplasties to examine the effect of acetabular component position. In group A, 38 of the components were implanted according to manufacture's instructions with all peripheral fins in contact with acetabular bone; as such, the acetabular components were in a relatively vertical position with a mean angle of inclination of 61.9 degrees. Three of these patients developed recurrent dislocations necessitating revision of the acetabular component. In group B, 37 hips, a more horizontal orientation was used despite the fact that all of the peripheral fins of the acetabular component did not engage acetabular bone; in this group the mean angle of inclination was 49.7 degrees. Only one of these hips recurrently dislocated and required revision. There were no problems in this group associated with provisional component stability caused by inadequate peripheral fixation. Radiographs of all patients were obtained at 4 years after surgery (range, 4.0-4.3 years). Pelvic osteolysis had occurred in 24% of hips in group A and 13% of group B. Asymmetric polyethylene wear was observed in 5.1% of the hips in group A; no hip in group B showed wear asymmetry. Acetabular component migration developed in 19% of group A hips and 5% of group B hips. The Mayo clinical hip score was excellent in both groups: group A 71/80, group B 73/80. At an intermediate follow-up it is clear that significant problems can be encountered when this component is positioned in a relatively vertical position to facilitate engaging all four peripheral fins in bone. We have addressed this problem by placing the cup in a more anatomic position of inclination while maintaining provisional rim fixation. This has resulted in a decreased incidence of pelvic osteolysis and fewer complications overall. PMID- 9726319 TI - Clinical results of the midstem porous-coated anatomic uncemented femoral stem in primary total hip arthroplasty: a five- to nine-year prospective study. AB - The clinical and radiologic results of an inclusive series of 60 patients (70 hips) who had primary total hip arthroplasty using the porous-coated anatomic (PCA) midstem femoral prosthesis was prospectively studied. The midstem component features a proximal circumferential porous bead coating similar to the PCA primary stem; but increased proximal thickness, increased length, and a distal anterior curve for additional rotational stability. The mean Harris Hip Score rose from 39.5 points before surgery to 91.3 points at a minimum follow-up of 5 years (average, 69 months); 88% were good or excellent. Moderate or severe thigh pain on a visual analogue scale was reported by 30% of cases, and was more common in women. Radiographic analysis indicated preservation of proximal bone stock and bony ingrowth in 87%, but stem subsidence in 9%. One stem has been revised for subsidence and thigh pain (1.4%), and one stem is radiographically loose, but the patient refuses surgical revision. Endosteal osteolysis was rarely seen (2.8%) and was benign in appearance. Acetabular components used included 63 nonmodular PCA metal-backed cups and 7 hemispherical porous ingrowth cups fixed with screws. One PCA cup was revised for loosening (1.4%), and one is radiographically loose but stable (1.4%). Only one cup exhibited an area of osteolysis. At this intermediate follow-up the clinical outcome of the midstem component is stable and excellent. The radiographic results appear superior to the PCA primary stem, with a lower incidence of stem subsidence and osteolysis. The prevalence of thigh pain is a concern and we recommend regular follow-up of patients with the midstem femoral implant, and the use of a visual analogue thigh pain scale when any femoral prosthesis is evaluated. PMID- 9726320 TI - Five-year clinical and radiographic follow-up of the uncemented long-term stable fixation total hip arthroplasty. AB - Thirty-nine uncemented porous-coated long-term stable fixation total hip prostheses with a minimum 5-year follow-up, were retrospectively reviewed for clinical and radiographic outcome. Clinical evaluation was performed using the modified Harris Hip Score. Additionally, all patients completed a satisfaction questionnaire. Anteroposterior view and lateral view radiographs were obtained and compared with immediate postoperative films utilizing the Hip Society radiographic evaluation form for uncemented implants. The follow-up period averaged 69 months (range, 60-87 months). At the most recent follow-up visit Harris Hip Scores averaged 88 points (range, 68-100 points), with 79% good or excellent results. Of the 8 hips (6 patients) with fair or poor results, 5 patients (6 hips) were Charnley category C patients. All patients were satisfied with their surgery and all, but 2 stated that their function had significantly improved. The incidence of significant thigh pain was 13%. Calcar osteolysis was present in 13 of 39 femurs with the majority of cases being minimal. All but one femoral component demonstrated bone ingrowth. There was no distal femoral osteolysis present and no femoral revisions have been performed or are planned. Acetabular osteolysis was present in 7 of 39 hips, with 4 of the 7 centered around acetabular fixation screws. All patients who had acetabular bone loss had some degree of femoral osteolysis. Thus far, one patient has required acetabular revision secondary to osteolysis. Acetabular osteolysis in this series was more profound than on the femoral side and two other patients are being considered for revision due to pelvic side osteolysis. In conclusion, the uncemented long-term stable fixation femoral component proved to be durable in this series of patients. The circumferential porous coating on the femoral implant may protect against distal osteolysis. A concerning rate of severe pelvic osteolysis and impending failure was noted and may lead to a greater need for revision surgery with longer follow-up. PMID- 9726321 TI - Influence of intramedullary versus extramedullary alignment guides on final total knee arthroplasty component position: a radiographic analysis. AB - A prospective study of 116 consecutive Kinemax cemented posterior cruciate ligament-retaining total knee arthroplasties was carried out. Similar surgical technique was used with a single variable: 61 were implanted using intramedullary guides on the tibia and 55 were implanted using extramedullary guides on the tibia. A radiographic study was performed after at least 1 year of follow-up to evaluate postoperative component position and compare the difference in the accuracy of positioning of the femoral and tibial components. Radiographic analysis showed that satisfactory position was achieved using both types of instrumentation. No statistically significant difference was observed in either the coronal or sagittal plane of the femoral component and the sagittal plane positioning of the tibial component. However, the coronal plane positioning of the tibial component revealed a statistically significant difference (P < .01), with intramedullary guides being superior to extramedullary guides. Also observed, was that using either technique, patients with less accurate postoperative positioning tended to be obese, with wide intramedullary canals. Patients with significant extraarticular deformities, marked bowing, and those with prior surgery or fractures may not be suitable for intramedullary guides, and they may require the use of extramedullary guides and intraoperative radiographic control. The ideal indication for the use of intramedullary instrumentation is in the patient who is not obese, with no extraarticular deformity, and with a well-defined, but not excessively wide, tibial medullary canal. Since tibial component malalignment in general, and coronal plane malalignment in particular, may adversely affect the long-term survival of total knee arthroplasties, the use of intramedullary alignment instrumentation is recommended when possible. PMID- 9726322 TI - An oblong revision cup for large acetabular defects: design rationale and two- to seven-year follow-up. AB - Loosening and migration of acetabular components often lead to extensive bony defects with an elongated, oval acetabular cavity. In these cases standard implants will not reestablish and maintain sufficient stability without leaving bone defects or using massive bone grafts or excess cement and additional metal rings or shells, disadvantages that are overcome by using an oblong revision cup without cement. The titanium shell is available in different sizes, is screwed to the autochthonous acetabular bone and houses an oblong polyethylene inlay, designed to reestablish the normal anatomic hip center. Of 109 consecutive revision cups, 102, implanted for American Academy of Orthopaedic Surgeons (AAOS) defects types I-IV, were followed up clinically and radiologically for 2 to 7 years (mean, 3.6 years). Primary stability was achieved in all cases. In 40% no bone grafting was necessary at all. The radiological follow-up revealed good remodeling of the surrounding bone and osseointegration of the implants. Zonal radiolucent lines, always smaller than 2 mm, were seen in 18 cases, only once completely and in only 5 cases partially progressing. Six cups migrated slightly (< or =2 mm), two moderately (3-5 mm), all without clinical symptoms, and two more than 5 mm. Migration and radiolucencies were mainly seen in patients with allografts and major defects, which indicates that bone ingrowth appears more unlikely in such cases. Few asymptomatic cases showed zonal sclerotic lines. There were two aseptic loosenings, one in a case with pelvic discontinuity, the other in a patient with severe rheumatoid arthritis following two previous revisions. Survivorship analysis based on implant removal because of aseptic loosening as the endpoint shows a cumulative success rate of 98.1% at 8 years. PMID- 9726323 TI - Differences in the rates of penetration determined from radiographic and shadowgraphic measurements of acetabular sockets. AB - A number of previous studies have observed a marked difference in the penetration depths recorded between direct and radiographic measurement, although no assessment of how these discrepancies affect the rates of penetration has been undertaken. In this report, the penetration depth of 95 sockets was determined from both prerevision radiographs and casts of the retrieved sockets using the uniradiographic and shadowgraphic techniques, respectively. It was observed that the mean discrepancy between the penetration rates derived from the two measurement methods was 0.046 (SE 0.017) mm yr(-1), which was significantly different from zero at P = .007. It was concluded that the penetration rates derived from this method of radiographic assessment seriously underestimated the true value as measured by a direct method by approximately 20%. As a consequence, the discrepancy in penetration rates between those sockets that are loose at revision surgery (direct measurement) and the general population of acetabular components (radiographic measurement) may not be as large as previously thought. PMID- 9726324 TI - Methyl methacrylate levels in unwashed salvage blood following unilateral total knee arthroplasty. AB - To evaluate the safety of autologous reinfusion of drain blood in total knee arthroplasty (TKA), eight patients were prospectively evaluated to quantify levels of methyl methacrylate (MMA) monomer in systemic blood, and in their drain blood after unilateral cemented TKA. The systemic blood was analyzed before and after reinfusion of the drain blood. The drain blood was analyzed before reinfusion, and both before and after filtration through a 40-microm filter. A separate study was performed on 10 patients to assess the effect of blood, time, and filtration on MMA levels. Levels of MMA monomer in salvage blood were low enough to allow safe reinfusion. Systemic blood showed no evidence of MMA monomer either before reinfusion of salvage blood or at 5 minutes after reinfusion. Elimination of MMA is dependent on the time that MMA is exposed to blood and is independent of filtration. PMID- 9726325 TI - Effect of the posterior cruciate ligament in knee-joint proprioception in total knee arthroplasty. AB - The primary purpose of the study was to examine the role of the posterior cruciate ligament (PCL) in knee-joint proprioception after total knee arthroplasty (TKA). Knee-joint proprioception was measured in 10 patients with nonsacrificed PCL TKAs and 10 with sacrificed PCL TKAs. Knee-joint proprioception was evaluated through reproduction of static knee angles using a Penny and Giles electrogoniometer. The primary variable was absolute angular error (AAE). AAE was defined as the absolute value of the difference between the test angle and the patient's perceived version of the test angle. Proprioception deficit was compared to the WOMAC questionnaire which evaluates pain, stiffness, and physical function of the lower extremity. No significant difference was found between the nonsacrificed PCL TKA (4.33 degrees +/- 1.52 degrees) and sacrificed PCL TKA (4.38 degrees +/- 1.39 degrees) AAE values (P > .4). Furthermore, no significant differences were observed in the WOMAC questionnaire scores for all three parameters between the two types of knee prosthesis (P > .35). The current findings suggest that the preservation of the PCL in TKA may not improve knee joint proprioception and subsequently may not improve TKA functional performance. PMID- 9726326 TI - Late sciatic nerve entrapment following pelvic plate reconstruction in total hip arthroplasty. AB - Delayed sciatic neuropathy due to pelvic reconstruction plate loosening following complex acetabular reconstruction in total hip arthroplasty seems not be have been previously reported. We identified a 79-year-old woman who developed progressive neurologic signs of entrapment 6 months following reconstruction of a pelvic discontinuity due to fracture nonunion caused by radiation necrosis. Magnetic resonance imaging of the lumbar spine was unrevealing and electromyography demonstrated a peripheral neurogenic process involving the sciatic nerve. Sciatic nerve exploration was done at 12 months after surgery finding a loose screw in the pelvic plate impinging the nerve. Substantial improvement in clinical symptoms resulted from removal and nerve release. PMID- 9726327 TI - Impingement after total knee arthroplasty caused by cement extrusion and proximal tibiofibular instability. AB - A 57-year-old patient with rheumatoid arthritis showed posterolateral impingement after total knee arthroplasty. The radiographs showed bone cement extrusion posterolateral to the tibial tray. Arthrotomy through a posterolateral approach revealed that the impingement was caused not only by cement extrusion against the fibular head but also by proximal tibiofibular joint instability. It was speculated that rheumatoid arthritis had caused proximal tibiofibular instability, active knee motion had caused fibular head shift by tension of biceps femoris and the fibular head had been impinged on the extruded cement. In cementing the tibial tray, especially in a rheumatoid patient, it is of paramount importance to take caution against posterolateral cement extrusion in order to minimize the risk of fibular head impingement during total knee arthroplasty. PMID- 9726328 TI - Osteolysis associated with cemented total knee arthroplasty. AB - Osteolysis has not been mentioned as a complication or cause of failure of cemented total knee arthroplasties in long-term follow-up studies. We are aware of a single case report of osteolysis after cemented total knee arthroplasty. We report the case of an 87-year-old woman with massive osteolysis beneath a cemented tibial component. PMID- 9726329 TI - Intraoperative pacemaker dysfunction caused by the use of electrocautery during a total hip arthroplasty. AB - Pacemaker dysfunction encountered during orthopedic procedures is a rare but potentially life-threatening complication. With an increasing number of orthopedic procedures performed on the aging population, it is not uncommon to encounter patients with pacemakers requiring major orthopedic intervention. Most, if not all, major orthopedic procedures performed today require the use of electrocautery for hemostasis. In this article we review the literature for pacemaker complications and report a case of pacemaker failure after a single use of the unipolar electrocautery on a patient undergoing a total hip replacement. PMID- 9726330 TI - Avulsion fracture of the ischium following complex total hip arthroplasty: an unusual cause of hip pain. AB - Total hip arthroplasty in the high riding dislocated hip is a technically difficult undertaking, with major reconstruction required on both the acetabular and femoral sides. With reconstruction at a near-anatomic hip center, reduction of the arthroplasty is difficult because of the long-standing limb shortening. The major block to reduction is tension of the soft tissues, particularly the hamstrings. We report a case of ischial tuberosity avulsion fracture following such a complex reconstruction despite femoral shortening subtrochanteric osteotomy. This illustrates the importance of the hamstring group in maintaining the dislocation and emphasizes the need to prevent overtension of the soft tissues in such complex reconstructive procedures. PMID- 9726331 TI - Review of thrombotic thrombocytopenic purpura in the setting of systemic lupus erythematosus. AB - OBJECTIVE: Thrombotic thrombocytopenic purpura (TTP) has been described in association with systemic lupus erythematosus (SLE) rarely. The diagnosis of TTP as a process separate from SLE may be difficult because both share similar features, including thrombotic microangiopathy. METHODS: A case is described of the simultaneous occurrence of TTP and SLE. The clinical, laboratory, and histologic findings of the patient are reported. The association of TTP and SLE in the literature is analyzed. We review separately the pathogenesis, role of antiphospholipid antibodies, and the differential diagnosis of TTP complicating the course of SLE. RESULTS: Forty cases of TTP in association with SLE are reported in the world literature. Three distinct groups were defined by the presentation of TTP that occurred subsequent to, before, or simultaneous with SLE (groups 1, 2, and 3, respectively). Renal biopsy in a patient with lupus nephritis may reveal thrombotic microangiopathy, which may be seen independently or represent a concomitant systemic thrombotic process such as TTP, disseminated intravascular coagulation, or antiphospholipid antibody syndrome. CONCLUSION: TTP in association with SLE is rare, and the diagnosis may be challenging. Although the etiology of TTP remains elusive, certain autoimmune mechanisms, platelet abnormalities, and fibrinolytic disorders may be shared with SLE and provide the basis for their association. Management requires timely diagnosis and aggressive treatment by therapeutic plasma exchange. PMID- 9726332 TI - Autoimmune thrombocytopenia in primary antiphospholipid syndrome and systemic lupus erythematosus: the response to splenectomy. AB - OBJECTIVE: To study the outcome of splenectomy in the management of thrombocytopenia. METHODS: Cases of systemic lupus erythematosus (SLE) or primary antiphospholipid syndrome (PAPS) complicated by severe thrombocytopenia were identified from a database of patients attending outpatients over the period 1978 to 1996. Clinical presentation, laboratory investigations, and response to medical treatment and/or splenectomy were documented. RESULTS: A total of 17 patients had severe thrombocytopenia; splenectomy was performed on 13: 9 with SLE, 3 with PAPS, and 1 with lupus-like disease (LLD). After splenectomy, six of nine patients with SLE and all three patients with PAPS gained complete remission of thrombocytopenia. There were no complications from splenectomy. The remaining four patients, three with SLE and one with PAPS, remained in a stable partial or full remission with oral medication and did not require splenectomy. CONCLUSION: This study re-emphasizes the place for splenectomy in SLE patients and supports its role in the management of thrombocytopenia in PAPS. PMID- 9726333 TI - Outcome of pregnancy in three patients with primary antiphospholipid syndrome after stroke. AB - OBJECTIVE: Ischemic stroke is the most common neurological manifestation in patients with antiphospholipid syndrome (APS). Pregnancy in APS patients markedly increases the risk of thrombosis. There is no data on pregnancy outcome in patients with APS with a history of an ischemic stroke. We report our experience with three APS patients with a history of stroke who had successful pregnancies and deliveries. PATIENTS: Three patients with APS and previous stroke were treated with small doses of aspirin and anticoagulants during pregnancy. RESULTS: The patients remained free of attacks of cerebral ischemia during their pregnancies and at follow-up periods of 1 to 4 years. CONCLUSIONS: Successful pregnancy and delivery is possible in APS patients with a history of stroke, treated with low-dose aspirin and anticoagulants. A previous episode of cerebral ischemia should not be considered an absolute contraindication for an APS patient to become pregnant. PMID- 9726334 TI - Coexisting Sjogren's syndrome and sarcoidosis in the lung. AB - CONTEXT: Sjogren's syndrome (SS) and sarcoidosis are diseases of unknown origin that are considered to result from abnormal regulation of the immune system. Pulmonary involvement by SS and sarcoidosis may have similar clinical and radiographic manifestations, making it difficult for the clinician to distinguish between these diseases. OBJECTIVES: This study was undertaken to analyze the characteristics of SS and sarcoidosis in the lung to identify distinguishing features that may assist clinicians in the differentiation of these conditions. DESIGN: We present two cases with severe pulmonary impairment in which the distinction between SS and sarcoidosis required lung tissue biopsy. The literature regarding the pulmonary manifestations of these diseases is reviewed. RESULTS: The clinical, pathological, radiographic, and physiological characteristics of lung disease in the setting of SS and sarcoidosis can be very similar, preventing a diagnosis solely on clinical grounds. This is exemplified in the two cases reported. In one patient who carried the diagnosis of sarcoidosis, examination of lung tissue revealed lymphocytic interstitial pneumonitis consistent with SS. In the other patient, who had previously been diagnosed with SS on clinical grounds, examination of lung tissue showed lymphocytic interstitial pneumonitis with scattered noncaseating granulomas, suggesting the possibility of coexisting SS and sarcoidosis. A literature review indicated that lung involvement by SS may be difficult to distinguish from that of sarcoidosis. Furthermore, several cases have been reported in which both diseases coexisted. CONCLUSIONS: Because SS and sarcoidosis may coexist and present with similar pulmonary manifestations, aggressive evaluation including tissue biopsy may be required. However, even tissue biopsy may not distinguish between these entities unless noncaseating granulomas are seen (in the case of sarcoidosis) or isolated lymphocytic interstitial pneumonitis is detected (in the case of SS). When both features (ie; noncaseating granuloma and lymphocytic interstitial pneumonitis) are encountered in the same organ, we believe these diseases are coexisting. Distinguishing both conditions may have prognostic implications, because sarcoidosis may present as an autolimiting process and frequently resolves spontaneously without significant residual functional impairment. In contrast, pulmonary involvement with SS often leads to permanent defects and may progress to incapacitating disease. PMID- 9726336 TI - Colchicine: 1998 update. AB - OBJECTIVE: To present an update of the use of colchicine in patients with familial Mediterranean fever (FMF) and other rheumatic and nonrheumatic diseases. DATA SOURCES: Published studies on colchicine retrieved from MEDLINE searches from 1987 to 1997 and reports presented at national and international meetings. STUDIES SELECTION AND EXTRACTION: All studies were reviewed by the authors. Reports addressing the topics of colchicine pharmacokinetics, biological effects, indications for use, and side effects were selected. DATA SYNTHESIS: Colchicine is an alkaloid that may interfere with microtubule formation, thereby affecting mitosis and other microtubule-dependent functions. It has a bioavailability of 25% to 50% when administered orally. Colchicine and its metabolites are excreted through the urinary and biliary tracts. It may be used while breast feeding; however, amniocentesis should be performed when used in pregnancy. The drug may be given to children with FMF. The efficacy of colchicine has been proved in FMF, gout, Behcet's disease, and cirrhosis. Its place in the treatment of scleroderma, sarcoidosis, and skin disorders remains to be determined. Gastrointestinal side effects occur early and are most common manifestations of colchicine toxicity. Severe colchicine toxicity results in multiple organ failure, convulsions, coma, and death. Potentially, effective treatment with Fab anti-colchicine antibodies unfortunately is unavailable; therefore, treatment is supportive. CONCLUSIONS: Colchicine is a relatively safe and effective medication for several disorders when used in appropriate dosage in patients with normal kidney and liver function. PMID- 9726335 TI - Dactylitis: implications for clinical practice. AB - OBJECTIVES: To assess the specificity of dactylitis for the diagnosis of spondyloarthropathy, sarcoidosis, and gout; and to characterize dactylitis specifically associated with gout. METHODS: Dactylitis was prospectively assessed among all individuals presenting to the Arthritis Center of Northeast Ohio from 1986 to 1996. RESULTS: Dactylitis was observed in 12% of individuals with spondyloarthropathy, 17% with sarcoidosis, and 5% with gout, but not in 96 patients with rheumatoid arthritis or in 2,434 patients with osteoarthritis, neck or back pain, or collagen vascular diseases. Among individuals with spondyloarthropathy, dactylitis was present in 22% with psoriatic, 28% with Reiter's syndrome, and only 7% with undifferentiated spondyloarthropathy. Gouty dactylitis was found only in individuals with polyarticular disease. CONCLUSIONS: Dactylitis is a valuable clue in the differential diagnosis of arthritis. Compared with the wider spectrum in children, sausage-shaped digits have only a few known causes in adults: Reiter's syndrome, psoriatic arthritis, sarcoidosis, flexor tendon sheath infections, and gout. In our series, the presence of dactylitis eliminated rheumatoid arthritis from the differential diagnosis. PMID- 9726337 TI - The role of inflammation in disk herniation-associated radiculopathy. AB - OBJECTIVE: The causes and physiopathology of low-back pain and acute lumbar radiculopathy remain unclear. A compression of the nerve root by protruded disk has been suggested but explains only partially the physiopathology of radicular pain. This article provides an overview of the role of inflammation in disk herniation-associated radiculopathy. METHODS: A review of the relevant literature in American and European medical journals was performed. RESULTS: Several studies have identified inflammatory mediators (phospholipase A2, prostaglandin E2, leukotrienes, nitric oxide, immunoglobulins, pro-inflammatory cytokines such as interleukin [IL]-1alpha, IL-1beta, IL-6, and tumor necrosis factor alpha [TNFalpha]) and autoimmune reaction (macrophages expressing IL-1beta, intercellular adhesion molecules) in disk herniation. An appealing hypothesis is that the leakage of these agents may produce an excitation of the nociceptors, a direct neural injury, a nerve inflammation, or an enhancement of sensitization to other pain-producing substances (such as bradykinin), leading to the nerve root pain. However, the role of these inflammatory mediators in the pathophysiology of lumbar radiculopathy has not been proven. Several findings suggest that this inflammatory response, which occurs in the early stage of disk herniation, is transient. Indeed, most studies of chronic disk herniation samples failed to demonstrate inflammation. CONCLUSION: Although inflammation may partially explain lumbar radiculopathy, involvement of inflammatory mediators in the physiopathology of disk herniation-associated radiculopathy has not been proven. PMID- 9726338 TI - Molecular tools for epidemiologic study of infectious diseases. PMID- 9726339 TI - Symptoms of acute otitis media. AB - BACKGROUND: The decision to seek medical advise for children during upper respiratory infections is largely based on the parental assumption that the child's symptoms are related to acute otitis media. The symptoms related to acute otitis media, however, are considered nonspecific. METHODS: Altogether 857 healthy day-care children (mean age, 3.7 years) were followed up for 3 months, and the symptoms of each child were compared during upper respiratory infections with and without acute otitis media. RESULTS: A total of 138 children had upper respiratory infections with and without acute otitis media. The symptom with the strongest association with acute otitis media was earache [relative risk (RR), 21.3; 95% confidence intervals (CI), 7.0 to 106, P < 0.0001] but sore throat (RR = 3.2; CI = 1.1 to 11; P = 0.027), night restlessness (RR = 2.6; CI = 1.1 to 6.9; P = 0.024) and fever (RR = 1.8; CI = 1.1 to 3.2; P = 0.025) also had significant associations. Logistic regression analysis showed 71% of the cases to be correctly diagnosed on the basis of the symptoms of earache and night restlessness. The parents were able to predict the presence of acute otitis media with a sensitivity and specificity of 71 and 80%, respectively (positive predictive value, 51%; negative predictive value, 90%). CONCLUSIONS: Despite the limited value of symptoms in differentiating acute otitis media from upper respiratory infection, the parents are able to predict acute otitis media somewhat reliably. More symptoms than have been reported earlier appeared to be associated with acute otitis media. PMID- 9726340 TI - Microbiology of acute otitis media in Costa Rican children. AB - BACKGROUND: Because of the increasing number of resistant middle ear pathogens reported from different centers worldwide, an active surveillance of the microbiology and susceptibility pattern of middle ear pathogens is required for proper antimicrobial recommendations among different regions of the world. OBJECTIVE: To study the microbiology and susceptibility pattern of middle ear pathogens obtained from Costa Rican children with acute otitis media. METHODS: Between 1992 and 1997 a diagnostic tympanocentesis was performed in 398 Costa Rican patients with acute otitis media. Middle ear fluid was obtained for culture and minimal inhibitory concentrations were determined by the E-test technique in those isolates obtained between October, 1995, and January, 1997. RESULTS: The most common pathogens cultured were Streptococcus pneumoniae (30%), Haemophilus influenzae (14%), Staphylococcus aureus (4%) and Streptococcus pyogenes (4%). Moraxella catarrhalis was uncommon. Beta-lactamase production was low (3.7%) among the H. influenzae isolates but frequent among the Staphylococcus aureus (57.1%) and M. catarrhalis (100%) strains. Overall 9 of 46 S. pneumoniae isolates (19.6%) exhibited decreased susceptibility to penicillin of which 8 isolates (17.4%) showed intermediate and one strain (2.2%) high level resistance. Among the penicillin-susceptible S. pneumoniae isolates, susceptibility to the following antimicrobials was: 81%, azithromycin; 89%, clarithromycin; and 100%, ceftriaxone and trimethoprim-sulfamethoxazole (TMP-SMX). Among the penicillin resistant S. pneumoniae isolates the percentage of susceptible strains was 89% for azithromycin, clarithromycin and ceftriaxone and 67% for TMP-SMX. CONCLUSIONS: Based on this microbiologic information the agents considered first line drugs in the treatment of acute otitis media in Costa Rica remain amoxicillin or TMP-SMX. PMID- 9726341 TI - Response to a heptavalent conjugate Streptococcus pneumoniae vaccine in children with recurrent infections who are unresponsive to the polysaccharide vaccine. AB - OBJECTIVE: To determine whether children with recurrent respiratory infections who failed to respond to the conventional polysaccharide vaccine would respond to a pneumococcal conjugate vaccine. METHODS: Children referred to our clinic for recurrent respiratory infections who had no known primary or secondary immunodeficiencies were immunized with a 23-valent pneumococcal polysaccharide vaccine. IgG antibodies to pneumococcal serotypes 1, 3, 4, 6B, 9V, 14, 18C, 19F and 23F were determined by enzyme-linked immunosorbent assay before and 4 to 6 weeks after immunization. An adequate IgG antibody response to an individual serotype was arbitrarily defined as a postimmunization antibody titer > or =1.3 microg/ml or at least 4 times the preimmunization value. Immunization with an experimental CRM197-heptavalent pneumococcal conjugate vaccine was offered to patients without an adequate response to 4 or more vaccine serotypes (nonresponders). Post-conjugate immunization antibody concentrations were measured 4 to 6 weeks later. RESULTS: In nonresponder patients (n = 17) geometric mean post-conjugate immunization (C) serum antibody concentrations (microg/ml) compared with post-polysaccharide (PS) concentrations were: (serotype, C vs. PS) 4, 1.11 vs. 0.30 (P = 0.000227); 6B, 0.46 vs. 0.20 (P = 0.017267); 9V, 0.82 vs. 0.29 (P = 0.002163); 14, 1.88 vs. 0.27 (P = 0.000615); 18C, 0.98 vs. 0.32 (P = 0.021962); 19F, 1.24 vs. 0.34 (P = 0.002844); and 23F, 0.87 vs. 0.16 (P = 0.000194). In responder patients (n = 67), after 1 dose of the polysaccharide vaccine, geometric mean antibody concentrations were: 4, 1.05; 6B, 0.96; 9V, 1.55; 14, 1.65; 18C, 1.62; 19F, 1.30; and 23F, 1.02. CONCLUSIONS: Our results show that a pneumococcal conjugate vaccine is capable of inducing an IgG response in patients with recurrent infections who had failed to mount an adequate response to the polysaccharide vaccine. Conjugate vaccines may be of value in the management of children with recurrent pneumococcal respiratory infections. PMID- 9726342 TI - Reevaluation of antipyretics in children with enteric fever. AB - BACKGROUND: The Committee on Infectious Diseases of the American Academy of Pediatrics recommends withholding antipyretic administration to patients with enteric fever because of the risk of shock developing as a consequence. OBJECTIVE: To evaluate the effects of antipyretics on blood pressure in children with enteric fever. METHODS: A retrospective review of medical records of patients admitted to Texas Children's Hospital from January, 1977, to October, 1997, with a diagnosis of enteric fever. All febrile episodes were evaluated for the use of antipyretics and evidence of hypotension or cardiovascular decompensation associated with them. RESULTS: Twenty-nine patients with enteric fever were identified. Salmonella typhi caused 23 of these infections. Antipyretics were used in all but one patient. We did not find any association between the use of antipyretics and the development of hypotension. One patient developed shock and adult respiratory distress syndrome >36 h after start of antibiotic therapy and unrelated to fever or antipyretic use. Two patients had evidence of dehydration. No other complications occurred. CONCLUSIONS: We did not find any complications associated with the use of acetaminophen or ibuprofen in children with enteric fever. The effects of antipyretics in enteric fever should be further studied. PMID- 9726343 TI - Albendazole therapy in children with focal seizures and single small enhancing computerized tomographic lesions: a randomized, placebo-controlled, double blind trial. AB - BACKGROUND: Single small enhancing computerized tomographic (CT) lesions (SSECTLs) are common in children with focal seizures. These are considered to represent solitary cysticercus granulomas. Controversy exists regarding their treatment. OBJECTIVE: To evaluate the efficacy of albendazole in cases of focal seizures with SSECTLs. DESIGN: Randomized, placebo-controlled, double blind trial. SETTING: Pediatric service of Nehru Hospital, PGIMER, an urban tertiary care teaching hospital. SUBJECTS: 63 children between 2 and 12 years of age with focal seizures for <3 months and SSECTLs. INTERVENTION: All children were randomly assigned to receive either albendazole (15 mg/kg/ day) or placebo for 28 days. CT scan was done at 1 and 3 months after beginning treatment. Codes opened after 6 months of inclusion in the study showed that 31 had received albendazole and 32 had received placebo. All children were followed up for at least 15 months. RESULTS: Disappearance of lesions on CT scan was noted in 41% of albendazole vs. 16.2% of placebo patients after 1 month of follow-up (P < 0.05) and 64.5% of albendazole- vs. 37.5% of placebo-treated patients after 3 months of follow-up (P < 0.05). During the first 4 weeks of therapy seizure recurrence was seen in 9.7% of albendazole vs. 3.2% of placebo-treated children (odds ratio, 3.32; 95% confidence interval, 0.33 to 33.8). After 4 weeks seizure recurrence was seen in 31.3% of placebo-treated children vs. 12.9% of albendazole-treated children (odds ratio, 3.07; 95% confidence interval, 1.18 to 11.15). CONCLUSIONS: Albendazole therapy results in significantly faster and increased resolution of solitary cysticercus lesions (SSECTLs) and appears to reduce the risk of late seizure recurrences. PMID- 9726344 TI - Pediatric visceral leishmaniasis in southern France. AB - PURPOSE: The purposes of this study were to describe the characteristics of pediatric visceral leishmaniasis in southern France and to evaluate a new scheme of therapy. METHODS: Hospital records of 59 children with visceral leishmaniasis were retrospectively reviewed. The period of the study was from 1981 to 1997. RESULTS: All children but one lived or had previously dwelled in the south of France. None was coinfected with human immunodeficiency virus or known to be immunocompromised. The mean age was 31 months; 10 children were younger than 1 year when admitted to the hospital. The male:female ratio was 0.73. Fever and splenomegaly were present in 90 and 100%, respectively. Anemia, leukopenia and thrombocytopenia were commonly observed, especially in the youngest patients. Hypergammaglobulinemia was noted in 64%. A biopsy sample of the bone marrow was always performed, but direct microscopic examination failed to identify Leishmania in 13 (22%) cases. In these patients specific serology and genomic amplification with polymerase chain reaction were useful tools for the diagnosis. All patients were initially treated with meglumine antimonate (Glucantime). Twenty-six (44%) patients receiving the drug experienced at least one adverse event during treatment. Treatment failure occurred in six children (10%), who were subsequently cured with liposomal amphotericin B. Three additional children were treated with liposomal amphotericin B. All the children were finally cured and no death was observed. CONCLUSION: Our experience suggests that liposomal amphotericin B is effective therapy for visceral leishmaniasis in children. PMID- 9726346 TI - A statewide survey of immunization rates in Minnesota school age children: implications for targeted assessment and prevention strategies. AB - BACKGROUND: A retrospective statewide immunization survey of the 69115 Minnesota children who entered kindergarten in 1992 was conducted. METHODS: Information was collected from school immunization records on date of birth, dates of vaccination for each dose of vaccine, address of residence and race/ethnicity (when available). Immunization rates were assessed retrospectively for each month of a child's life from 2 to 48 months of age. Age-appropriate immunization was defined as receipt of all scheduled vaccines within 30 days of the recommended age. RESULTS: Immunization levels varied by vaccine, age of the child and race/ethnicity. For example at 19 months of age, 73% of students had received measles, mumps, rubella vaccine; however, only 39% had received their fourth dose of diphtheria, tetanus and pertussis vaccine. White, non-Hispanic students consistently had higher vaccination rates than children of other racial/ ethnic groups. For example 45% of white, non-Hispanic students were age-appropriately vaccinated at 16 months of age compared with 25% of Blacks, 30% of American Indians, 30% of white Hispanics and 28% of Asian-Pacific Islanders (Mantel Haenzel chi square, P < 0.001 for each comparison). Furthermore coverage rates frequently varied significantly by neighborhood, thereby identifying pockets of underimmunization within communities. CONCLUSION: Our data demonstrate that vaccination rates can vary substantially by age, race/ ethnicity and neighborhood. Detailed immunization assessment is necessary so that effective targeted interventions can be developed. PMID- 9726345 TI - Value of cerebrospinal fluid cytochemical examination for the diagnosis of congenital toxoplasmosis at birth in France. AB - BACKGROUND: Because of routine screening and treatment of pregnant women for Toxoplasma infection in France, most neonates born to mothers who seroconverted during pregnancy are either not infected or asymptomatic. Early diagnosis relies mainly on radiologic, ophthalmologic and biologic tests. Cerebrospinal fluid (CSF) cytochemical evaluation is one of several tests performed in parallel to increase the overall sensitivity of the diagnostic evaluation. Our goal was to assess the value of cytochemical examination and to confirm whether using a portion of available CSF for this analysis is legitimate. METHODS: The individual performance of each of the two cytochemical tests and their combined value when used in parallel were assessed. These findings were then compared with the anti Toxoplasma IgM and IgA serum titers and the clinical, ophthalmologic and radiologic findings at birth. RESULTS: CSF cytochemical analysis was possible in only 52% of the 233 children in the study. Our results in 112 children indicated poor sensitivity estimates. There was no significant change in the posttest probability of infection compared with the pretest estimation of risk in cases of a negative finding. After a mean follow-up of 80 months there was no evidence that CSF cytochemistry helped predict the risk of sequelae. CONCLUSION: In our setting cytochemical examination did not significantly contribute to the diagnosis of congenital infection at birth. Because of the limited quantity of CSF available, we suggest the use of other methods with higher yield. PMID- 9726347 TI - Outbreak of Acinetobacter spp. bloodstream infections in a nursery associated with contaminated aerosols and air conditioners. AB - BACKGROUND: Acinetobacter spp. are multidrug-resistant bacteria that grow well in water and cause infections with unexplained, increased summer prevalence. In August, 1996, eight infants acquired Acinetobacter spp. bloodstream infection (A BSI) while in a nursery in the Bahamas; three infants died and an investigation was initiated. METHODS: A case patient was defined as any newborn in the nursery during August 6 to 13, 1996, with A-BSI. To identify risk factors for A-BSI we conducted a retrospective cohort study and performed environmental cultures and air sampling using settle plates. The genetic relatedness of environmental isolates was assessed by pulsed field gel electrophoresis. RESULTS: Of 33 patients in the nursery 8 (24%) met the case definition. Patients with peripheral iv catheters were more likely to develop A-BSI (8 of 21 vs. O of 10, P < 0.05). Multivariate analysis among patients with iv catheters indicated that only exposure to one nurse was an independent risk factor for developing A-BSI (P < 0.005). Nursery settle plates were more likely to grow Acinetobacter spp. than were settle plates from other hospital areas (8 of 9 vs. 0 of 5, P < 0.005); cultures from nursery air conditioners also grew Acinetobacter spp. Environmental isolates were genetically diverse. After installation of a new air conditioner in May, 1995, A-BSIs occurred more frequently during months of increased absolute humidity or environmental dew point. CONCLUSIONS: Acinetobacter spp. may cause nosocomial BSI and death among infants during periods of polyclonal airborne dissemination; breaks in aseptic technique during i.v. medication administration may facilitate transmission from the environment to the patient. Environmental conditions that increase air conditioner condensate may predispose to airborne dissemination via contaminated aerosols and increase the risk of nosocomial A BSI. PMID- 9726348 TI - A prospective study of astrovirus diarrhea of infancy in Mexico City. AB - AIM: To describe the epidemiologic and clinical characteristics of astrovirus associated diarrhea in a cohort of young children from a periurban community in Mexico City. METHODS: From November, 1988, through December, 1991, a total of 214 children were enrolled in a longitudinal study of diarrhea and monitored from birth to 18 months of age. A stool specimen was collected during each episode of diarrhea. Specimens from a total of 510 diarrhea episodes were tested for astrovirus by enzyme immunoassay and examined for other enteric pathogens. The antigenic types of astrovirus were determined by a typing enzyme immunoassay. RESULTS: Astrovirus was detected in 26 (5%) of 510 diarrhea episodes, with an incidence rate of 0.1 episode/child year; the highest rate was in children 13 to 18 months of age. Astrovirus-associated diarrhea was characterized by a median of 4 stools (range, 2 to 10) during the first 24 h, a median duration of 3 days (range, 1 to 21), vomiting (20%), and fever (7%). No cases of dehydration or repeat symptomatic infections were observed. Coinfection with another pathogen was detected in 11 of the 26 episodes (42%). Serotype 2 (35%) was most common, followed by serotypes 4 (15%), 3 (11%), and 1 and 5 (4% each); 31% were nontypable. Astrovirus-associated diarrhea was less severe, as measured by the number of stools (4.3 +/- 1.9), than diarrhea caused by rotavirus (7.1 +/- 2.8) or when coinfections occurred (5.5 +/- 1.6; P = 0.008). CONCLUSIONS: Astrovirus was associated with 5% of the episodes of diarrhea in this cohort of young Mexican children and presented as a mild secretory diarrhea. Five predominant antigenic types were detected with type 2 being the most common. PMID- 9726349 TI - Early experience with protease inhibitors in human immunodeficiency virus infected children. PMID- 9726350 TI - Off label use of antimicrobial agents in infants, children and adolescents: a time for action. PMID- 9726351 TI - Community-acquired methicillin-resistant staphylococcus aureus infections. PMID- 9726352 TI - Management of vaginal candidiasis: a review. PMID- 9726353 TI - Strategies to promote appropriate antimicrobial use. PMID- 9726354 TI - Hantavirus pulmonary syndrome treated with inhaled nitric oxide. PMID- 9726355 TI - Neuroimaging findings in children perinatally infected with the human immunodeficiency virus. PMID- 9726356 TI - Effect of superimposed infections on viral replication in human immunodeficiency virus type 1-infected children. PMID- 9726357 TI - Invasive Kingella kingae infection associated with stomatitis in children. PMID- 9726358 TI - Laryngeal tuberculosis. PMID- 9726359 TI - Neonatal hand abscess, osteomyelitis and meningitis caused by Streptococcus pneumoniae. PMID- 9726360 TI - Thrombocytopenia, hepatitis and pleural effusion in a three-month-old Mexican boy. PMID- 9726361 TI - Circumcision and infectious diseases. PMID- 9726362 TI - Transplacental measles immunity. PMID- 9726363 TI - Should we use vancomycin as prophylaxis to prevent neonatal nosocomial coagulase negative staphylococcal septicemia? PMID- 9726364 TI - Modification of superoxide dismutase (SOD) mRNA and activity by a transient hypoxic stress in cultured glial cells. AB - In order to understand the role of superoxide dismutase (SOD) in response to transient hypoxia or hypoxia-reperfusion in astrocytes, the present study performed an in vitro investigation using rat glial cells in culture. Hypoxia was induced by an incubation with nitrogen gas for 10 min and that followed a further reperfusion with air for 10 min was indicating as hypoxia-normoxia. Activity of SOD was determined by the reduction of nitroblue tetrazolium (NTB). Changes of mRNA for Cu,Zn-SOD or Mn-SOD were also characterized using Northern blotting analysis. Transient hypoxia increased the activity of Mn-SOD but not that of Cu,Zn-SOD in glial cells. Expression of mRNA for SOD was also elevated in cells received hypoxia and the mRNA level for Mn-SOD raised higher than that for Cu,Zn SOD. In cells received hypoxia-reperfusion, these changes of SOD both the activity and the mRNA level were not observed. Otherwise, the SOD protein amount, both Cu,Zn-SOD and Mn-SOD, identified by Western blotting was not changed in glial cells receiving hypoxic stress or not. The obtained results suggest that gene expression and activity of Mn-SOD in glial cells can be activated in response to the transient hypoxic stress. PMID- 9726365 TI - Paracrine production of nerve growth factor during rat dorsal root ganglion development. AB - Nerve growth factor is a target derived growth factor. In the peripheral nervous system, it is produced by tissues innervated by the sympathetic nervous system and small sensory neurons. In the present study, we tested the hypothesis that an alternate source of nerve growth factor must be available to support dorsal root ganglion neurons before they make connection with the target. Using reverse transcriptase polymerase chain reaction (RT-PCR), we detected nerve growth factor mRNA at embryonic day 12 to 17, but not in adult dorsal root ganglia. In situ hybridization studies revealed positive staining in satellite/supportive cells juxtaposed to dorsal root ganglion neurons. Our study suggests that nerve growth factor from supporting cells may have a paracrine function during development of primary sensory neurons. PMID- 9726366 TI - Potential role for imidazole in the rhythmic respiratory activity of the in vitro neonatal rat brainstem. AB - We used the imidazole-binding agent, diethylpyrocarbonate (DEPC), to test the hypothesis that rhythmic respiratory activity of the in vitro neonatal rat brainstem-spinal cord preparation was functionally dependent on imidazole. Neural activity was recorded from spinal nerves (C1-C4) during superfusion with 95%O2/5%CO2 buffer at pH 7.3 and T = 26 degrees C. Superfusate containing DEPC (40 mM) caused cessation of rhythmic activity within minutes. In eight of 33 preparations, microinjection of DEPC (32 nmol) onto the ventral medullary surface (VMS) reduced burst amplitude by at least 50% within 10 min, and in 12 of 33 preparations, microinjection of DEPC produced neural apnea. Therefore, we conclude that proteins containing imidazole near the VMS are critically important for the maintenance of rhythmic respiratory activity in vitro. Furthermore, alphastat regulation of respiration may be an essential trait of this preparation. PMID- 9726367 TI - Rat oligodendrocyte O-2A precursor cells and the CG-4 oligodendrocyte precursor cell line express cadherins, beta-catenin and the neural cell adhesion molecule, NCAM. AB - Oligodendrocyte precursor cell migration throughout the developing central nervous system (CNS) and cessation of migration are poorly understood but are likely to involve cell adhesion molecules. The expression and distribution of neural cell adhesion molecule (NCAM), cadherins and beta-catenin were investigated in the CG-4 cell line and primary rat oligodendrocyte progenitor cells (O-2A) by immunofluorescence and Western blotting. NCAM was expressed by both cell types and was found all over the surface of both CG-4 cells and O-2A progenitor glia. The presence of a cadherin was detected in both CG-4 cells and O 2A progenitor glia, and this molecule was distributed all over the cell body and cell processes at different stages of differentiation. Beta-catenin showed a very similar distribution to that of the cadherin. We conclude that CG-4 cells are a valid model system to study cell adhesion molecule expression and function in oligodendrocyte progenitor cells. PMID- 9726368 TI - Evidence for NOS-containing renal neuronal somata transiently expressing a catecholaminergic phenotype during development in the rat. AB - Transiently catecholaminergic cells (TC-cells) expressing tyrosine hydroxylase (TH) have been shown in a variety of tissues during embryonic life. To investigate the possible relationship of nitric oxide synthase (NOS)-containing renal neuronal somata (RNS) and the TC-cells, we examined serial 100 microm slices of whole kidneys for TH-immunofluorescence and NADPH-d histochemistry during prenatal and postnatal development. The number of TH-cells increased during the prenatal period, peaked at birth and were very rare by PD21. A subpopulation of TH-immunoreactive RNS displayed NADPH-d activity. By PD21 the TH positive RNS had practically disappeared while the number of NADPH-d positive RNS was markedly increased. These results suggest that kidneys possess transient catecholaminergic cells which display NOS-activity and that NOS expression may be the end-point in the differentiation of the RNS. PMID- 9726369 TI - Beta1 integrin deficiency impairs migration and differentiation of mouse embryonic stem cell derived neurons. AB - Cell-matrix interaction plays an important role during neuronal development, which is demonstrated by comparing wild type (D3)- and beta1 integrin-deficient (G201) embryonic stem cell derived neurons. In D3 preparations complex networks of functionally coupled neurons with bi- and multipolar morphologies develop. In contrast, neuronal differentiation is retarded in G201 derived neurons, recognised by limited migration and restricted morphological differentiation. Furthermore, beta1 integrin deficiency causes a delay in expression of major neurotransmitters like GABA and glutamate as well as of synaptophysin. These findings indicate a prominent role of beta1 integrin for both morphological and chemical differentiation. PMID- 9726370 TI - Sciatic and femoral nerve sensory neurones occupy different regions of the L4 dorsal root ganglion in the adult rat. AB - The topographical distribution of sciatic and femoral nerve sensory neuronal somata in the L4 dorsal root ganglion of the adult rat was mapped after retrograde tracing with one or two of the dyes Fast Blue, Fluoro-Gold, or Diamidino Yellow. The tracers were applied to the proximal transected end of either nerve alone, or from both nerves in the same animal using separate tracers. Three-dimensional reconstructions of the distribution of labelled neurones were made from serial sections of the L4 dorsal root ganglion which is the only ganglion that these two nerves share. The results showed that with little overlap, femoral nerve neurones distribute dorsally and rostrally whereas sciatic nerve neurones distribute medially and ventrally. This finding indicates the existence of a somatotopical organisation for the representation of different peripheral nerves in dorsal root ganglia of adult animals. PMID- 9726372 TI - Performance of visually triggered wrist movements task in monkey: an application of information theory to evaluate deficits following unilateral substantia nigra pars reticulata lesion. AB - On the basis of an information theory approach, visual-motor efficiency (VME) was analyzed during pointing movement tasks in rhesus monkeys. This application was used to evaluate deficits in the ability to perform pointing movements as well as to chart the progress of the monkey during the chronic stage recovery period following unilateral lesion of the medial part of the substantia nigra pars reticulata. This unilateral lesion produced slight but significant chronic contralateral head roll tilt associated with body turning and induced profound modification of the VME at the beginning of the chronic stage of the symptomatology. Recovery developed in the following weeks, leading to a 3-fold increase in VME. However, at the end of the test (4 weeks postoperative), performance remained sub-normal: a decrease of 20-30% when compared to the control. PMID- 9726371 TI - Effect of acute exposure to 3-nitropropionic acid on activities of endogenous antioxidants in the rat brain. AB - The response of endogenous antioxidants to acute exposure of the mitochondrial inhibitor, 3-nitropropionic acid (3-NPA), was investigated in selected rat brain regions. Rats treated with 3-NPA (30 mg/kg, s.c.) were sacrificed at 30, 60, 90 and 120 min after injection to examine the alterations in reduced glutathione levels (GSH), and activities of antioxidant enzymes, superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) in the hippocampus (HIP), frontal cortex (FC), and caudate nucleus (CN). CAT activity increased in the HIP 90 min after 3-NPA treatment. While cytosolic copper/zinc SOD (CuZn-SOD) and mitochondrial manganese SOD (Mn-SOD) levels increased in the FC at 120 min, only the Mn-SOD increased in the CN 90 min after treatment. The activity of GPx decreased in the HIP 120 min after 3-NPA injection. When compared with the control, administration of 3-NPA led to GSH depletion in HIP within 120 min. The depletion of GSH and induction of antioxidant enzyme activities after the 3-NPA exposure suggest conditions favorable for oxidative stress. PMID- 9726373 TI - Labeling of N-acetylaspartate and N-acetylaspartylglutamate in rat neocortex, hippocampus and cerebellum from [1-13C]glucose. AB - Both N-acetylaspartate (NAA) and N-acetylaspartylglutamate (NAAG) are localized almost exclusively to neurons, and have become important markers of neuronal viability in a number of cerebral pathological conditions. Using nuclear magnetic resonance spectroscopy combined with [1-13C]glucose administration (200 min infusion) we show that the synthesis of both NAA and NAAG can be observed. Label was incorporated into NAA from labeled acetate and from labeled aspartate, while NAAG was labeled from labeled glutamate. The low fractional enrichment of NAA (ca. 3%) relative to aspartate (20%) suggests a slow turnover rate, while NAAG (20.0%) and glutamate (25.2%) labeling were nearly equal, suggesting that NAAG labeling is near steady state. The rapid turnover of NAAG suggests an important role in glutamate delivery, while the slow rate of NAA turnover implies that its major role is as substrate for the formation of NAAG. PMID- 9726374 TI - Localisation of the nitric oxide (NO)/cGMP-pathway in the vestibular system of guinea pigs. AB - The exact distribution of nitric oxide-synthases (NOS) in the vestibular system has not been described satisfying yet. Immunostaining, using specific antibodies to the three known NOS-isoforms, to cyclic guanosine monophosphate (cGMP) and soluble guanylyl-cyclase (sGC), the second messenger system of nitric oxide (NO), was performed on paraffin sections of temporal bone from guinea pigs. eNOS could be detected in vestibular ganglion cells and in nerve fibres, including the calyces, surrounding the type 1 hair cells (HC). bNOS was found in the sensory epithelium, ganglion cells and in bone, while iNOS could not be found. NOS detection was accompanied by reactivity to sGC and to cGMP. This finding implies that b- and eNOS-generated NO is involved in regulative processes in neurotransmission and regulation of blood flow. PMID- 9726375 TI - IL-10 reduces rat brain injury following focal stroke. AB - The effects of the anti-inflammatory cytokine, IL-10, on brain injury following permanent focal ischemia were determined. Rats subjected to occlusion of the right middle cerebral artery (MCAO) were administered IL-10 (1 microg) centrally into the lateral ventricle 30 min and 3 h post MCAO or systemically into the tail vein (5 or 15 microg/h) starting 30 min post MCAO for 3 h. Brains were removed 24 h later and infarct size was measured. IL-10 administered centrally significantly (P < 0.01) reduced infarct size by 20.7% +/- 6.0 compared to vehicle. Systemic IL 10 administration at 5 and 15 microg/h significantly (P < 0.05) decreased infarct size (40.3% +/- 14.0 and 30.7% +/- 13.7, respectively). These studies indicate that an anti-inflammatory therapeutic approach using IL-10 can provide neuroprotection in ischemic stroke. PMID- 9726376 TI - Local application of collagen containing brain-derived neurotrophic factor decreases the loss of function after spinal cord injury in the adult rat. AB - We studied the effect of local application of brain-derived neurotrophic factor (BDNF) on functional recovery after dorsal spinal cord transection in the adult rat. BDNF was applied at the site of the lesion in rat tail collagen type I. Locomotion was measured for 4 weeks using the BBB locomotor rating scale. One day after injury and application of BDNF the performance of treated rats was significantly increased as compared to controls (BBB-score 11.5+/-1.3 (mean +/- SEM) and 7.5+/-1.3, respectively). This difference remained significant during the first week. Histological examination of the spared spinal cord tissue at the lesion centre 4 weeks after lesioning showed no significant difference between control and BDNF-treated animals. The results indicate that local application of BDNF results in a decreased loss of function in the partially transected rat spinal cord starting one day after injury. PMID- 9726377 TI - Regenerating axons of the rat require a local source of proteins. AB - Regenerating axons need proteins to grow and we explored whether a local supply is necessary. Crushed peroneal nerves were entubulated with silicone sleeves, plain or loaded with cycloheximide (CHX); some nerves were frozen to kill resident cells. When a plain sleeve was placed distal to the crush, axons regrew 5.0 mm in 3 days (pinch test), and 4.6 mm when the sleeve was placed around a frozen nerve (n.s). CHX administered distal to the crush reduced the elongation by approximately 58% (P < 0.01) in unfrozen or frozen nerves whilst its administration central to the crush was ineffectual. Immunostaining of nerves with GAP-43 gave similar values. Under the electron microscope, axonal sprouts were less frequent when CHX was used irrespective of the cellular or acellular condition of the nerve. Therefore, an inhibitor of protein synthesis reduces axonal regrowth, an effect mediated neither by parent neurones nor by resident cells. We propose that axons synthesize proteins. PMID- 9726378 TI - Adenosine A1 and A2A receptor antagonists stimulate motor activity: evidence for an increased effectiveness in aged rats. AB - The motor effects of selective adenosine A1 and A2A receptor antagonists were tested in young (2 months) and aged (24 months) Wistar rats. In young rats, both the selective A2A receptor antagonist 5-amino-7-(2-phenylethyl)-2-2(2-furyl) pyrazolo[4,3-e]-1,2,4-triazo++ + lo[1,5-c]pyrimidine (SCH 58261, minimal effective dose 2 mg/kg intraperitoneally (i.p.)) and the selective A1 receptor antagonist 8-cyclopentyl-1,3-dimethylxanthine (CPT, minimal effective dose 1.2 mg/kg i.p.) stimulated motor activity. In old rats, both compounds induced significant motor activation starting from doses lower than those required in young animals. Specifically, the minimal effective doses of SCH 58261 and CPT in aged rats were 1 and 0.6 mg/kg i.p, respectively. The results indicate that both adenosine A1 and A2A receptors play a functional role in the control of motor activity, and, therefore, the blockade of both receptor subtypes is involved in the motor stimulating properties of methylxanthines. Also the evidence indicates, for the first time, that in aged animals the motor inhibitory adenosinergic tone seems to be increased with respect to young animals. PMID- 9726379 TI - Filamentous alpha-synuclein inclusions link multiple system atrophy with Parkinson's disease and dementia with Lewy bodies. AB - Alpha-synuclein forms the major component of Lewy bodies and Lewy neurites, the defining neuropathological characteristics of Parkinson's disease and dementia with Lewy bodies. Here we show that alpha-synuclein is also the major component of the filamentous inclusions of multiple system atrophy which comprises several neurodegenerative diseases with a shared filamentous pathology in nerve cells and glial cells. These findings provide an unexpected link between multiple system atrophy and Lewy body disorders and establish that alpha-synucleinopathies constitute a major class of human neurodegenerative disorder. PMID- 9726380 TI - Review article: the diagnosis and treatment of haematinic deficiency in gastrointestinal disease. AB - Deficiency of any of the vitamins and minerals essential for normal erythropoiesis (haematinics), including iron, copper, cobalt, vitamins A, B12, B6, C, E, folic acid, riboflavin and nicotinic acid, may be associated with defective erythropoiesis and anaemia. Iron, vitamin B12 and folate are the haematinics for which deficiency states manifest most often clinically and are the focus of this review. The normal absorption of these haematinics and gastrointestinal causes of their deficiency are described. Investigations, including the use of homocysteine metabolite levels and new techniques such as serum transferrin receptor assays, and treatment of haematinic deficiency are discussed in detail. PMID- 9726381 TI - Increased bone resorption in patients with Crohn's disease. AB - BACKGROUND: Patients with Crohn's disease are at risk of osteoporosis and premature fracture. However, the pathophysiology underlying bone loss remains poorly understood and the optimum treatment has not been established. AIM: To investigate mechanisms of bone loss in Crohn's disease using biochemical markers of bone turnover. METHODS: Bone mineral density was measured at the hip and spine using dual-energy X-ray absorptiometry in 117 patients (48 male) with Crohn's disease. Bone turnover was assessed by measuring serum osteocalcin (BGP), pro collagen carboxy-terminal propeptide (PICP), bone specific alkaline phosphatase (BALP) and urinary deoxypyridinoline (DPD); and compared to age-matched healthy controls (n = 28). RESULTS: Bone mineral density was reduced (z-score < -1) in 48 (41%) patients with Crohn's disease. Mean values for bone formation markers in patients with Crohn's disease were all within the normal reference range (BGP 8.92 (+/- 3.23) ng/mL (normal range 3.4-10.0), BALP 17.6 (+/- 12.6) U/L (normal range 11.6-43.3), PICP 95.1 (+/- 46.5) ng/mL (normal range 69-163)) and were not significantly different to the control population. However, mean urinary DPD was significantly higher in patients with Crohn's disease compared to healthy controls (10.97 (+/- 9.22) nM DPD/mM creatinine vs. 5.02 (+/- 1.03) nM DPD/mM creatinine, difference in means = 5.95, 95% CI: -9.6 to -2.3, P = 0.00001) and compared to the UK reference range DPD levels were increased in 74 (63%) patients. CONCLUSIONS: Bone resorption as evidenced by urinary DPD was frequently increased in patients with Crohn's disease and was significantly higher than in an age-matched control population. The high levels of urinary DPD suggest increased bone collagen degradation may contribute to osteoporosis in patients with Crohn's disease. These results suggest anti-resorptive agents such as the bisphosphonates may be effective treatment for osteoporosis in Crohn's disease. PMID- 9726382 TI - Olsalazine versus mesalazine in the treatment of mild to moderate ulcerative colitis. AB - AIM: To compare the efficacy and tolerability of olsalazine sodium with enteric coated mesalazine in inducing endoscopic remission in patients with mild to moderate active ulcerative colitis. PATIENTS AND METHODS: Patients with mild to moderate active ulcerative colitis were randomized to receive either olsalazine sodium, 3 g/day (n = 88), or mesalazine, 3 g/day (n = 80), for up to 12 weeks. RESULTS: Of the patients treated with olsalazine sodium, 52.2% achieved endoscopic remission, compared with 48.8% of patients treated with mesalazine. This difference was not significant (P = 0.67). There was a nonsignificant trend for patients with left-sided colitis or a more severe endoscopic grade to achieve remission if they were treated with olsalazine sodium than if they were treated with mesalazine. Both treatments were comparable with respect to clinical activity index and an investigator's global assessment. Seventy patients reported one or more adverse events; adverse events were seen in 45% of olsalazine sodium treated patients and in 36% of mesalazine-treated patients. Eleven patients treated with olsalazine sodium and nine patients treated with mesalazine withdrew from the study because of adverse events. One patient treated with olsalazine sodium compared with two treated with mesalazine stopped treatment because of diarrhoea. Serious adverse events occurred in three patients treated with olsalazine sodium and in four treated with mesalazine. CONCLUSION: Therapeutic effectiveness and tolerance to the treatment did not differ between olsalazine sodium, 3 g/day, and mesalazine, 3 g/day, in inducing endoscopic remission in patients with mild to moderate active ulcerative colitis within 12 weeks of treatment. PMID- 9726383 TI - Interferon alpha with ribavirin for the treatment of chronic hepatitis C in non responders or relapsers to interferon monotherapy. AB - BACKGROUND: A more effective therapy for chronic hepatitis C virus-infected patients is needed. AIM: To evaluate the efficacy, tolerance and timing of response to interferon alpha plus ribavirin in 60 patients with no response or reactivation after interferon alpha alone. METHODS: Sixty patients, 42 non responders and 18 relapsers, received 3 million units three times weekly of interferon alpha-2b plus 1-1.2 g ribavirin daily, for 6 months. Basal biochemical and virological (HCV RNA and genotype) parameters were determined. Clinical examination, recording adverse effects, and laboratory tests, including viraemia, were carried out at 1, 2, 3 and 6 months. RESULTS: A significant (P < 0.001) progressive decrease of HCV RNA and alanine transaminase (ALT) levels was observed during treatment. On finalizing the sixth month, 42 patients (70%) had normal ALT and 26 (43.3%) were HCV RNA negative. Of these 26 complete responders, in 20 the viraemia was undetectable by the third month, while a late clearance at the sixth month of treatment was observed in six patients. Response rates were higher in previous responders to interferon alone (P < 0.05). Mild adverse effects appeared in 46 patients (79.6%), but only three were withdrawn due to serious side-effects. Significantly (P < 0.001), haemoglobin and leucocytes decreased, and bilirubin, ferritin and uric acid increased in the first month of treatment, with no changes thereafter. CONCLUSIONS: Interferon alpha plus ribavirin progressively decreased HCV RNA and ALT levels, achieving a complete response in the six months of treatment in 26 (43.3%) patients. This combined therapy was well tolerated. PMID- 9726385 TI - Short-term low-dose pantoprazole-based triple therapy for cure of Helicobacter pylori infection in duodenal ulcer patients. AB - BACKGROUND: The eradication of Helicobacter pylori infection has been achieved using various therapy regimens, but the efficacy of the proton-pump inhibitor pantoprazole as part of these regimens has not yet been widely tested. AIM: To evaluate the efficacy and tolerability of a 1-week low-dose pantoprazole-based triple therapy in patients with H. pylori-positive duodenal ulcer. METHODS: In an open single-centre prospective study, 71 patients with endoscopically proven active duodenal ulcer and H. pylori infection received pantoprazole 40 mg o.m. for 4 weeks, and during the first week a combination antimicrobial treatment comprising tinidazole 500 mg b.d. plus clarithromycin 250 mg b.d. H. pylori eradication was defined as concordant negative histology and rapid urease test performed at endoscopy 4-6 weeks after the end of treatment, confirmed 4 weeks later by 13C-urea breath test. RESULTS: Sixty-six patients (93%) completed the trial and five patients were lost to follow-up. H. pylori infection was cured in 61 out of the 66 patients who completed the trial (per-protocol analysis: 92.4%, 95% CI: 83.2-97.5%; intention-to-treat analysis: 85.9%, 95% CI: 75.7-93.0%). At final endoscopy, 65 out of 66 patients had healed ulcer (98.5%). Mild adverse events occurred in six patients (9.1%). CONCLUSIONS: One-week low-dose pantoprazole-based triple therapy is a simple, effective and well-tolerated regimen for ulcer healing and H. pylori eradication in patients with duodenal ulcer. PMID- 9726384 TI - One-week use of ranitidine bismuth citrate, amoxycillin and clarithromycin for the treatment of Helicobacter pylori-related duodenal ulcer. AB - BACKGROUND: Proton pump inhibitors have been widely used in combination with amoxycillin, clarithromycin or metronidazole for the treatment of Helicobacter pylori infection. AIM: To study the effects of 1-week ranitidine bismuth citrate (RBC)-based triple therapy in the treatment of H. pylori-related duodenal ulcers. METHOD: Patients with duodenal ulcers and H. pylori infection were prospectively randomized to receive either RBC with amoxycillin and clarithromycin for 1 week (RAC), or omeprazole with amoxycillin and clarithromycin for 1 week (OAC). No additional ulcer healing drug was used after the 1-week medication. Patients were assessed for H. pylori eradication, ulcer healing and side-effects after receiving the therapies. RESULTS: One hundred consecutive patients were recruited to this study, with 50 patients randomized to each treatment group. In the intention-to-treat analysis, duodenal ulcers were completely healed in 45 (90%) patients in the RAC group and 43 (89.6%) in the OAC group (P = 1.0). H. pylori eradication was confirmed in 47 (94%) in the RAC group and 42 (87.5%) in the OAC group (P = 0.31). There was no significant difference in the severity of side effects experienced by the two treatment groups. CONCLUSION: One-week RBC-based triple therapy is an effective treatment for H. pylori-related duodenal ulcers. The therapeutic effects are comparable to a 1-week course of proton pump inhibitor-based triple therapy. PMID- 9726386 TI - Changes in Helicobacter pylori-induced gastritis in the antrum and corpus during and after 12 months of treatment with ranitidine and lansoprazole in patients with duodenal ulcer disease. AB - BACKGROUND: Several studies have shown that acid-suppressing treatment leads to aggravation of Helicobacter pylori gastritis in the corpus. It remains unclear whether this augmentation of the inflammation reverts to baseline after termination of treatment. METHODS: In 114 H. pylori-infected duodenal ulcer patients we investigated the gastritis parameters in antral and corpus mucosa before treatment, after 6 and 12 months of therapy, and 6 months after termination of treatment with 15 mg lansoprazole or 150 mg ranitidine/day. RESULTS: Lansoprazole and ranitidine led to a significant aggravation of gastritis in the corpus after 6 and 12 months of treatment. However, while there was no change in gastritis in the antrum with ranitidine, treatment with lansoprazole led to partial elimination of H. pylori with improvement of the inflammation in this part of the stomach. Following termination of therapy, the observed changes reverted to baseline. No increase in intestinal metaplasia and/or atrophy in the antrum or corpus was observed. CONCLUSION: Both substances are associated with an aggravation of H. pylori gastritis in the corpus. However, only lansoprazole leads to a partial elimination of H. pylori with improvement of the inflammation in the antrum. The changes provoked by acid-suppressing treatment revert to baseline after termination of therapy. PMID- 9726387 TI - Incidence of duodenal ulcer healing after 1 week of proton pump inhibitor triple therapy for eradication of Helicobacter pylori. The Lansoprazole Helicobacter Study Group. AB - BACKGROUND: A number of clinical studies have assessed the efficacy of short-term twice-daily Helicobacter pylori eradication regimens but few have investigated the proportion of patients in whom duodenal ulcer disease was healed with these regimens. AIM: To compare the safety and efficacy of four 1-week H. pylori eradication regimens in the healing of H. pylori associated duodenal ulcer disease. METHODS: Following endoscopic confirmation of duodenal ulcer disease and a positive CLO test, patients underwent a 13C-urea breath test to confirm H. pylori status. Treatment with one of four regimens: LAC, LAM, LCM or OAM, where L is lansoprazole 30 mg b.d., A is amoxycillin 1 g b.d., M is metronidazole 400 mg b.d., C is clarithromycin 250 mg b.d., and O is omeprazole 20 mg b.d., was assigned randomly to those patients who were H. pylori positive, with 62 (LAC), 64 (LAM), 61 (LCM) and 75 (OAM) patients in each treatment group. Follow-up breath tests and endoscopies were performed at least 28 days after the end of treatment. RESULTS: Duodenal ulcer disease was healed 28 days after treatment in 53/62 (85.5%) patients who were treated with LAC, 52/64 (81.3%) of patients treated with LAM, 49/61 (80.3%) of patients treated with LCM and 60/75 (80.0%) of patients treated with OAM (intention-to-treat analysis, n = 262, assumed unhealed if no follow-up endoscopy was performed). All the treatments were of similar efficacy (P = 0.85, chi-squared test) with regard to the healing of duodenal ulcer disease. CONCLUSIONS: The four 1-week treatment regimens were equally effective in healing H. pylori associated duodenal ulcer disease. PMID- 9726388 TI - Effects of cholinergic agents on anorectal physiology. AB - BACKGROUND: Despite their potential therapeutic benefit, the effects of cholinergic agents on anal function have been poorly investigated. AIM: To analyse the effects of neostigmine and atropine on anorectal responses to rectal isobaric distension. METHODS: This was a placebo-controlled, randomized, double blind crossover study, performed in 12 healthy volunteers who received intravenously, on 3 separate days, neostigmine, atropine or the placebo. During each day of the experiment, seven pressure steps (ranging from 1 to 31 mmHg) in three different protocols of rectal isobaric distension (phasic, stepwise and tonic) were applied using an electronic barostat. Manometric responses of the anal canal, adaptative volumes and perception scores of the rectum were recorded. RESULTS: During stepwise distension, a significant drug effect was encountered at the anal level. No drug effect was observed on the other investigated parameters (rectal volumes and rectal perception scores) or for the other modes of distension. Compared to placebo, neostigmine significantly decreased pressures at the upper level of the anal canal for both recto anal inhibitory reflex and mean resting pressures. In contrast, atropine significantly increased pressures at the lower part of the anal canal but did not modify upper anal pressures. CONCLUSION: The present study suggests that cholinergic effects result more from an indirect action on intermediate neurotransmitters and rectal myenteric neurons, than from a direct action on anal targets. PMID- 9726389 TI - The effect of cisapride on dyspepsia symptoms and the electrogastrogram in patients with non-ulcer dyspepsia. AB - BACKGROUND: The electrogastrogram (EGG), which records gastric myoelectrical activity, is abnormal in one-third of adult patients with non-ulcer dyspepsia (NUD). AIM: To observe the effects of cisapride on EGG in adults with NUD. METHODS: Twenty-seven NUD patients who had undergone a pre- and post-prandial EGG were entered into an open study. All patients completed a dyspepsia symptom questionnaire and were then treated with cisapride 10 mg t.d.s. The dyspepsia questionnaire was repeated in all those completing 4 weeks of treatment. Those with an initial abnormal EGG (< 70% of slow wave activity at 2-4 cycles per minute) had a repeat EGG at the end of the study. RESULTS: Treatment with cisapride was associated with a significant improvement in the post-prandial EGG (P = 0.007). After 4 weeks of treatment, 7 of 13 abnormal EGGs normalized. Symptom scores improved significantly in the 13 patients with an abnormal EGG who completed treatment (P < 0.0003), but not in NUD patients with a normal EGG (P = 0.48). CONCLUSION: In this open study, treatment of NUD with cisapride was associated with significant symptom improvement in patients with an abnormal pre treatment EGG, but not those with a normal EGG, with a significant improvement of the post-prandial EGG. PMID- 9726390 TI - The influence of cisapride on gastric tone and the perception of gastric distension. AB - BACKGROUND: Delayed gastric emptying, impaired gastric accommodation to a meal and hypersensitivity to gastric distension have been implied in the pathophysiology of functional dyspepsia. Dyspeptic patients are often treated with the prokinetic drug cisapride. AIM: To assess the effects of cisapride on perception of gastric distension and gastric accommodation to a meal. METHODS: Eighteen healthy volunteers underwent a gastric barostat study on two occasions, after pretreatment with placebo or cisapride 10 mg q.d.s. Graded isobaric and isovolumetric distensions were performed until the subjects reported discomfort. Volume and pressure changes were recorded and perception was scored by a questionnaire. In 10 volunteers, the amplitude of the gastric accommodation to a mixed liquid meal was also measured. RESULTS: Pre-treatment with cisapride significantly lowered thresholds for perception and for discomfort, both during isobaric (4.3 +/- 0.7 vs. 3.2 +/- 0.7 and 12.2 +/- 1.2 vs. 9.2 +/- 0.9 mmHg above minimal distending pressure (MDP), respectively, P < 0.05) and isovolumetric (256 +/- 46 vs. 200 +/- 35 and 644 +/- 36 vs. 511 +/- 40 mL, respectively, P < 0.05) distensions. Cisapride significantly enhanced the size of the meal-induced fundus relaxation (143 +/- 37 vs. 270 +/- 50 mL, P < 0.05). CONCLUSIONS: Cisapride enhances both the perception of gastric distension and the gastric accommodation to a meal. These data suggest that cisapride may provide benefit to patients with impaired postprandial relaxation of the fundus. PMID- 9726391 TI - Activation of genes for spasmolytic peptide, transforming growth factor alpha and for cyclooxygenase (COX)-1 and COX-2 during gastric adaptation to aspirin damage in rats. AB - BACKGROUND: NSAIDs, such as aspirin (ASA), cause widespread mucosal damage, but repeated ASA insults appear to induce mucosal tolerance (adaptation) to this injury. The mechanism of the gastric adaptation to the damage induced by ASA has not been fully explained. AIM: To determine the role of the mucosal gene expression for spasmolitic peptide (SP) (a member of trefoil peptides) and transforming growth factor alpha (TGF alpha) as well as for cyclooxygenase (COX) 1 and COX-2 during gastric adaptation to ASA in rats. METHODS: Gastric lesions were produced by ASA (100 mg/kg in 1.5 mL of 0.2 M HCl) applied intragastrically (i.g.) as a single dose. every day for 5 days. Control rats were given 1.5 mL of vehicle (0.2 M HCl i.g.) as a single dose, during 5 consecutive days. Gastric blood flow (GBF) was measured by H2-gas clearance technique and gastric mucosal specimens were taken for the assessment of cell proliferation rate in gastric mucosa by bromodeoxyuridine (BrdU) uptake, mucosal generation of prostaglandin E2 measured by radioimmunoassay, and for expression of SP, TGF alpha COX-1 and COX-2 mRNA as determined by RT-PCR. To quantify the relative amounts of mRNA for SP and TGF alpha, southern blotting analysis of the PCR products was performed and the intensity of PCR products was compared with that of beta-actin used as a standard. RESULTS: ASA applied once produced numerous gastric erosions, but with repeated ASA doses the adaptation to this NSAID developed, the area of gastric lesions being reduced by 86% after six consecutive ASA insults. This adaptation to ASA was accompanied by approximately a 90% reduction in prostaglandin E2 biosynthesis, by a significant rise in BrdU uptake by glandular cells predominantly in the neck region of gastric glands and by expression of SP (SP/beta-actin ratio; 0.96 +/- 0.08 in ASA-adapted mucosa vs. 0.38 +/- 0.05 in the control mucosa) and TGF alpha (TGF alpha/beta-actin ratio: 0.97 +/- 0.07 in ASA-adapted mucosa vs. 0.77 +/- 0.06 in the control mucosa). COX-1 expression was detected in vehicle-control gastric mucosa and after single exposure to ASA or after six consecutive ASA insults, while COX-2 mRNA was not detected in vehicle control gastric mucosa, but appeared after single ASA insult and was sustained after subsequent ASA doses. CONCLUSIONS: (i) Gastric adaptation to aspirin injury involves enhanced cell proliferation which appears to be mediated by increased expression of SP and TGF alpha, and (ii) rapid upregulation of COX-2 expression following single and repeated ASA insults may represent a compensatory response to suppression of prostaglandin generation by this NSAID. PMID- 9726392 TI - Relationship between fundic endocrine cells and gastric acid secretion in hypersecretory duodenal ulcer diseases. AB - BACKGROUND: Acid hypersecretion is associated with duodenal ulcer disease in the following conditions: Zollinger-Ellison syndrome (ZES) and antral gastrin cell hyperfunction (AGCH) due to hypergastrinaemia, or hypersecretory duodenal ulcer (HDU) without hypergastrinaemia. AIM: To evaluate whether quantitative changes in fundic ECL and D cells may be involved in acid hypersecretion. PATIENTS AND METHODS: Seven ZES, six AGCH and six HDU Helicobacter pylori-positive patients were compared. Basal (BAO) and pentagastrin-stimulated gastric acid secretions (PAO), and morphometry of fundic ECL and D cells were performed. The six AGCH and six HDU patients were investigated again using the same tests 1 year after H. pylori eradication. RESULTS: Median PAO values were no different in all the hypersecretory conditions studied. The median volume density of ECL cells in ZES was significantly higher than in controls (2.75, range 1.74-5.8 vs. 0.73, 0.52 1.11: P < 0.05), whereas it was in the control range in AGCH and HDU patients (0.77, range 0.20-1.39 and 0.99, range 0.42-1.51; respectively). The count of fundic D cells was significantly lower in AGCH patients than in all other investigated groups (median 0.16, range 0.1-0.52; P < 0.05). Cure of infection in AGCH and HDU patients did not modify the ECL cell volume density, whereas a significant increase in the count of fundic D cells was observed in AGCH patients. Thus, the ECL/D cell index was significantly affected in AGCH patients (P < 0.05), being higher during H. pylori infection (median 6, range 0.7-9.25) than after the cure (median 2.12, range 1.10-3.5). BAO and PAO were not affected by H. pylori eradication in either group. CONCLUSIONS: The study provides evidence, for the first time, that quantitative alterations in the fundic endocrine cells are not involved in acid hypersecretion of patients with hypersecretory states, and that eradication of H. pylori does not restore normal acid secretion values. PMID- 9726393 TI - Comparison of rabeprazole 20 mg vs. omeprazole 20 mg in the treatment of active gastric ulcer--a European multicentre study. The European Rabeprazole Study Group. AB - BACKGROUND: Rabeprazole sodium is the newest member of a class of substituted benzimidazole molecules known as proton pump inhibitors. Other proton pump inhibitors have been shown to be effective in healing active, benign gastric ulcers. METHODS: In this randomized, double-blind, multicentre study, conducted at 25 European sites, rabeprazole and omeprazole were compared in patients with active gastric ulcers. Two hundred and twenty-seven patients with active benign gastric ulcer were randomized to receive either rabeprazole 20 mg (n = 113) or omeprazole 20 mg (n = 114) once daily for 3 or 6 weeks, with healing monitored by endoscopy. RESULTS: After 3 weeks, complete healing (ITT analysis) was documented in 58% of patients given rabeprazole and 61% in patients given omeprazole (N.S.). After 6 weeks the healing rates were identical in both groups at 91%. Rabeprazole treated patients had numerically greater symptom relief at all 12 points of comparison. The differences significantly favoured rabeprazole at week 3 for daytime pain improvement (P = 0.023) and at week 6 for pain frequency (P = 0.006) and complete resolution of night pain (P = 0.022). Both drugs were well-tolerated over the 6-week treatment course. Mean changes from baseline to end-point in fasting serum gastrin were comparable. No significant differences in laboratory parameters were seen. CONCLUSION: In this study, rabeprazole produced healing rates comparable to omeprazole at weeks 3 and 6, but provided more consistent and occasionally significantly superior symptom improvement. Both treatments were well-tolerated. PMID- 9726395 TI - The Dornier Lithotripter U/15/50: a multifunctional and multidisciplinary workstation. AB - Since February 1, 1996, we have been using the Dornier Lithotripter U/50 in our Stone Center, first and foremost for SWL treatment of urinary lithiasis. In a limited series of 136 patients treated from February 1, 1996, through April 30, 1996, we obtained encouraging results: the average retreatment rate for stones of all sizes was 13.2%, and the stone-free rate at 3 months was 90.4%. We performed auxiliary endourologic procedures in only 5.9% of patients (2.2% before and 3.7% after SWL). Thus, we obtained an Effectiveness Quotient of 77. These results are compared with those in the literature. Apart from SWL treatments in urology, the system was also used for shockwave treatments in orthopedics, for endourologic procedures, and for diagnostic imaging. Because of the encouraging results in SWL and its excellent performance in multifunctional and multidisciplinary use, we consider the Dornier U/50 to be the best lithotripter we have worked with to date. PMID- 9726394 TI - Trends in prescribing H2-receptor antagonists and proton pump inhibitors in primary care. AB - BACKGROUND: H2-receptor antagonists and proton pump inhibitors account for approximately 15% of primary care prescribing costs in the UK. AIM: To examine the use of antisecretory drugs in primary care between October 1991 and September 1996. METHOD: Analysis of prescribing data from an ongoing postal survey performed every 3 months on a rolling quota of 250 UK general practitioners (GPs), identified from a representative sampling frame of 1000 GPs. RESULTS: There were 8811 new courses of proton pump inhibitors and 11,948 new courses of H2-receptor antagonists during this study. The number of new prescriptions for proton pump inhibitors increased by 174.5%, but decreased for H2-receptor antagonists by 12.5%. Proton pump inhibitors were mostly prescribed for reflux disease (52.7%) and H2-receptor antagonists for non-specific dyspepsia (43.6%). Proton pump inhibitors (14.1%) were less likely to be stopped than H2-receptor antagonists (35.3%) overall, and they were less likely to be stopped because of perceived ineffectiveness (5.3%) than H2-receptor antagonists (23.8%). The rate of stopping treatment because of side-effects was about 3% for both classes of drug. CONCLUSIONS: Prescribing of proton pump inhibitors has increased sharply each year since 1991. One reason may be that GPs perceive proton pump inhibitors to be more effective than H2-receptor antagonists. PMID- 9726396 TI - Intracorporeal or extracorporeal lithotripsy for distal ureteral calculi? Effect of stone size and multiplicity on success rates. AB - Over a period of 57 months, 404 patients with distal ureteral calculi were treated by in situ SWL on a Storz Modulith SL 20 lithotripter and 163 by ureteroscopy (URS) and Swiss Lithoclast stone fragmentation. The case notes on these patients were reviewed for comparison of the initial stone number and individual length and for the calculation of the stone-free, treatment, retreatment, secondary procedure, and complication rates. Complete data were available on 447 patients. The median stone length was 7.0 (range 4-25) mm in the SWL group and 8.0 (range 5-13) mm in the URS group. The single-treatment stone free rates for the SWL and URS groups were 74.8% and 89.7%, respectively, for single stones and 50.0% and 88.9%, respectively, for multiple (>1) stones. The mean treatment rates for the SWL and URS groups were 1.97 and 1.03, respectively, for single stones and 2.83 and 1.00, respectively, for multiple stones. The mean treatment rate for single stones subjected to SWL increased with increasing stone length (1.57 for stones <8 mm and 2.38 for stones >8 mm), whereas this was not the case for patients submitted to URS (1.20 and 1.27, respectively). The re treatment rate for each group showed a reciprocal trend. Of the SWL group, 25.9% of the patients eventually required URS to render them stone-free. Nearly all (96%) of the patients undergoing SWL were treated as outpatients. The mean hospitalization in the URS group was 1.1 days. Three patients who underwent URS sustained a ureteral perforation, which was managed successfully by double-J stent insertion. The ideal primary treatment for small (<8 mm) distal ureteral calculi is in situ SWL, with URS plus Lithoclast fragmentation being reserved for failed SWL, single stones >8 mm in length, and multiple stones. PMID- 9726397 TI - Ureteroscopic and radiographic imaging of the upper urinary tract. AB - Direct endoscopic viewing of the upper urinary tract offers multiple benefits, including recognition and diagnosis, and also permits unsurpassed accuracy in positioning working instruments for biopsy and treatment. We have demonstrated the normal and abnormal endoscopic findings within the upper urinary tract in patients who were treated ureteroscopically for numerous indications, including calculi, filling defects, or ureteral obstruction. Small (7 to 7.5F) rigid and flexible ureteroscopes were employed in all patients. Selected endoscopic images of the subject lesions were photographed using a 35-mm camera with an endoscopic zoom lens and ISO-400 film. Endoscopic findings are illustrated in 13 patients, with case summaries in 11. Selected radiographic and endoscopic images show the essential features in each case and demonstrate the value of ureteroscopic examination. PMID- 9726398 TI - Organ retrieval systems for endoscopic nephrectomy: a comparative study. AB - Laparoscopic or retroperitoneoscopic interventions such as nephrectomy or tumor nephrectomy call for the removal of large quantities of tissue, which can no longer be extracted via the relatively confined lumen of a cannula. For this purpose, a variety of organ retrieval systems have been designed and are commercially available with the aim of safe tissue retrieval. This paper summarizes the results of an experimental and clinical comparison of the most important organ entrapment systems suitable for endoscopic nephrectomy. The LapSacs was the first organ bag especially designed for laparoscopic nephrectomy. Despite various new modifications of this entrapment system, it still represents one of the best alternatives and has been used worldwide with success. However, because of its simplicity, it requires a certain laparoscopic expertise and involves a learning curve. Newly developed retrieval systems (i.e., LapBag, Extraction Bag, Endo-Catch) offer some advantages regarding the handling of the bag, which may be particularly useful during retroperitoneoscopic nephrectomy with a restricted working space. Retrieval systems (i.e., Endobag, Endopouch) with low resistance to tearing forces or permeability to tumor cells or bacteria (i.e., Espiner Bag) cannot be recommended for endoscopic nephrectomy. PMID- 9726399 TI - Balloon catheter dilatation in the treatment of ureteral and ureteroenteric stricture. AB - Balloon catheter dilatation is a low-cost alternative to open surgery in patients with ureteral strictures, leading to low morbidity and short hospitalization. The goal of this study was to evaluate the results of this technique in patients with inflammatory ureteral strictures or ureteroenteric strictures after radical cystectomy. Twenty-five ureteral strictures in 20 (15 male, 5 female) patients were consecutively treated by high-pressure balloon dilatation: 14 cases of ureteroenteric stricture (9 after ileal cutaneous diversion, and 5 after orthotopic enterocystoplasty) and 11 of ureteral stricture from various inflammatory causes (tuberculosis, iatrogenic injury, radiation therapy, parasitosis). Dilatation was performed by an antegrade (ureteroenteric strictures) or retrograde (inflammatory strictures) approach using a balloon insufflated up to 10 to 20 atm for 5 to 15 minutes. The ureter was stented for a mean time of 2.1 months (range 1-5 months). Results were evaluated clinically and radiologically (intravenous urogram or CT scan). Immediate success was assessed by intraoperative radiologic monitoring. Long-term success was defined as the absence of recurrence of the stenosis after 6 months. Nineteen procedures were successful among the 23 evaluable cases. With a mean follow-up of 16 months (range 6-39 months), the long-term success rate was 52%: 40% in ureteral strictures and 61% in ureteroenteric strictures. Five strictures secondary to cutaneous diversion and six caused by radiation therapy recurred after dilatation. After cutaneous diversion, the failure occurred mostly at the anastomosis and involved the crossed-over ureter. This study shows that high pressure balloon dilatation of ureteral strictures has a high early success rate and a long-term success rate of 52%. It can therefore be considered as an alternative to open surgery. PMID- 9726400 TI - Ureteroscopic lithoclast lithotripsy: a cost-effective option. AB - Seventy-four consecutive cases of ureteral stones listed for ureteroscopic lithotripsy were studied prospectively. In all cases, the Wolf 7.5F or 9F ureteroscope was used in conjunction with the Swiss Lithoclast system. Dormia baskets were employed on four occasions to prevent forward propulsion of fragments. Ureteroscopic access to the stones was successful in 70 patients (95%). Lithoclast lithotripsy was successfully applied in 68 patients (92%), with complete fragmentation noted in 62 patients (91%), one requiring two sessions. The 6-week stone-free rate was 96% for these patients. Five patients with partial fragmentation had successful adjuvant SWL. The overall successful fragmentation rate was thus 84% (62 of 74) and 91% (67 of 74) in combination with adjuvant SWL. Cost analysis indicated a three-fold advantage for the Lithoclast over Candela lasertripsy. Ureteroscopic Lithoclast lithotripsy is a cost-effective treatment modality for ureteral stones. PMID- 9726401 TI - Laparoscopic nephroureterectomy for upper tract transitional-cell cancer: technical aspects. AB - Standard surgical therapy for most patients with upper tract transitional-cell carcinoma (TCC) is total nephroureterectomy with excision of an ipsilateral cuff of bladder; this procedure is performed through two separate or one long abdominal incision. The laparoscopic approach has the same goals. At Washington University, our approach has been similar to the open operation in that the procedure is performed transperitoneally and a cuff of bladder is secured. Herein, our current method for laparoscopic nephroureterectomy is described and illustrated in detail. In addition, other laparoscopic techniques for performing the nephrectomy portion and handling the distal ureter are described. Overall, this technique is still evolving as laparoscopic surgeons attempt to balance the goal of a thorough nephroureterectomy with the need to make the procedure cost effective. PMID- 9726402 TI - Laser division of intraluminal sutures. AB - Neodymium:YAG and holmium:YAG lasers were used to remove intraluminal sutures from three patients. In two, the suture was in the bladder and had served as a nidus for stone formation. In the other, the suture was in the ureter. An in vitro study showed that the Nd:YAG laser could divide all types of suture readily, whereas the Ho:YAG laser could divide all materials except Gortex. These cases illustrate another application for lasers. PMID- 9726403 TI - Supracostal percutaneous nephrolithotomy for upper pole caliceal calculi. AB - The incidence of upper pole calculi is 15% of all caliceal calculi. The management of such calculi has been simplified since the advent of extracorporeal shockwave lithotripsy (SWL). In our experience, however, there is a subset of upper pole caliceal calculi wherein certain features can render SWL less than adequate treatment, namely diameter >1.5 cm, narrowing of the caliceal infundibulum, either singly or combined, and morbid obesity. In such instances, percutaneous nephrolithotomy (PCNL) is indicated. Percutaneous access to an upper pole calix can be difficult by a subcostal track. The supracostal 12th rib approach provides direct and efficient access to an upper pole calix and is ideally suited for upper pole calculi. Twenty-one patients with large or complex upper pole calculi were treated by supracostal PCNL. The maximum diameter of the calculi ranged from 7 to 40 mm. Eight were branched (staghorn). There was one horseshoe kidney, and calculi were bilaterally represented in another patient. Two patients were morbidly obese. All procedures were performed in one stage under general anesthesia. Following cystoscopy and ureteral catheterization, the upper pole calix was accessed directly with the aid of C-arm fluoroscopy and retrograde ureteral contrast injection. The percutaneous tract was dilated to a maximum of 26 F, a working sheath was inserted, and the calculi were extracted after ultrasonic or pneumatic fragmentation. One patient required secondary SWL for residual fragments. There were no intrathoracic complications, and blood loss was minimal. Large or complex upper pole caliceal calculi, particularly in the morbidly obese, can be treated effectively by PCNL using supracostal percutaneous access. PMID- 9726404 TI - Renal tuberculosis presenting as lateralizing hematuria diagnosis by ureteronephroscopy and selective upper tract urine culture. AB - A 48-year-old man presented with painless left lateralizing macroscopic hematuria. Ureteroscopy revealed flocculent material in the left kidney and white linear streaks on some upper pole papillae; urine culture from the left renal pelvis was positive for tuberculosis, whereas voided urine cultures were negative. Renal tuberculosis should be included in the differential diagnosis of lateralizing hematuria, especially in the absence of an obvious cause for the bleeding. Direct culture of urine from the renal pelvis may have more sensitivity than culture of voided urine in this circumstance. PMID- 9726405 TI - Percutaneous removal of stone from caliceal diverticulum in patient with nephroptosis. AB - A novel method is introduced for percutaneous stone extraction from a lower pole caliceal diverticulum in a patient with nephroptosis, also known as a floating kidney. The patient was fully recovered and asymptomatic at 2 months postoperatively with her kidney fixed in the flank position. PMID- 9726406 TI - Renal effects of carbon dioxide insufflation in rabbit pneumoretroperitoneum model. AB - To determine the effects of carbon dioxide insufflation on renal function in a pneumoretroperitoneum model, 24 adult New Zealand rabbits were divided into four groups, six rabbits in each. The first group underwent a 2-hour CO2 insufflation at a pressure of 10 mm Hg in the retroperitoneal space after balloon dissection. In another group, the same procedure was maintained for 4 hours. In the sham treated groups, the procedure was similarly carried out but without CO2 insufflation. In all four groups, serum and urine creatinine concentrations and renal artery and renal vein blood flow rates were determined separately at the beginning and at the end of the procedure and at 24 hours. Urine output was also recorded at the end of the procedure and at 24 hours. The serum creatinine in the 2- and 4-hour study groups had increased significantly at the end of the procedure, accompanied by a significant decrease in the urine creatinine value. Renal artery and renal vein blood flow rates and urine output were reduced in both groups during the study. All changes in the serum and urine creatinine, renal artery and vein flow rates, and urine output was more pronounced in the 4 hour group. All measures returned to their prestudy values by 24 hours. Pneumoretroperitoneum causes reversible renal dysfunction, which becomes more pronounced with prolonged insufflation. Further research is needed to show the impact of our findings in high-risk patients undergoing retroperitoneoscopic surgery. PMID- 9726407 TI - Transient cavitation and acoustic emission produced by different laser lithotripters. AB - Transient cavitation and shockwave generation produced by pulsed-dye and holmium:YAG laser lithotripters were studied using high-speed photography and acoustic emission measurements. In addition, stone phantoms were used to compare the fragmentation efficiency of various laser and electrohydraulic lithotripters. The pulsed-dye laser, with a wavelength (504 nm) strongly absorbed by most stone materials but not by water, and a short pulse duration of approximately 1 microsec, induces plasma formation on the surface of the target calculi. Subsequently, the rapid expansion of the plasma forms a cavitation bubble, which expands spherically to a maximum size and then collapses violently, leading to strong shockwave generation and microjet impingement, which comprises the primary mechanism for stone fragmentation with short-pulse lasers. In contrast, the holmium laser, with a wavelength (2100 nm) most strongly absorbed by water as well as by all stone materials and a long pulse duration of 250 to 350 microsec, produces an elongated, pear-shaped cavitation bubble at the tip of the optical fiber that forms a vapor channel to conduct the ensuing laser energy to the target stone (Moss effect). The expansion and subsequent collapse of the elongated bubble is asymmetric, resulting in weak shockwave generation and microjet impingement. Thus, stone fragmentation in holmium laser lithotripsy is caused primarily by thermal ablation (drilling effect). PMID- 9726408 TI - Necrosis of the urethra and tissue sloughing: a delayed complication after transurethral microwave thermotherapy. AB - Transurethral microwave thermotherapy is a minimally invasive treatment for benign prostatic hyperplasia designed to destroy hyperplastic tissue without damaging the urethra. We present an unexpected complication of prostatic urethral necrosis and tissue sloughing after thermotherapy and discuss its possible cause. PMID- 9726409 TI - Combination of pneumatic lithotripsy and transurethral prostatectomy in bladder stones with benign prostatic hyperplasia. AB - We present our experience with combined pneumatic lithotripsy and transurethral resection of the prostate (TURP) in 52 patients with bladder stone(s) and benign prostatic hyperplasia (BPH). All stones were fragmented with the pneumatic Swiss Lithoclast. Pneumatic lithotripsy and evacuation caused a mean increase of 16 minutes in operating time. No complications, other than mild hematuria, were observed intraoperatively because of pneumatic lithotripsy. We observed early and long-term complications related to the procedure in 13% of patients. The average hospital stay was 3.2 days. The combination of pneumatic lithotripsy and TURP appears to be an effective, safe, and economical treatment method for patients with bladder stone(s) and BPH. PMID- 9726410 TI - Uncommon complications of permanent stents. AB - The UroLume endoprosthesis has been used for recurrent bulbar urethral strictures, benign prostatic hyperplasia, and detrusor-external sphincter dyssynergia. Complications of the UroLume have been described by the North American Multicenter Trial as migration, encrustation, hyperplastic tissue growth, in addition to pain and irritative voiding symptoms. Generally, complications have been minimal, and few of the stents required removal. Patients with bulbar strictures were felt to be at relatively greater risk of hyperplastic tissue reaction if their stricture was secondary to trauma or if they had a prior urethroplasty. On the other hand, patients with benign prostatic hyperplasia were at risk of developing complications if they had median lobe enlargement or a short (<2.5 cm) prostatic urethra. We describe two cases in which off-label compassionate use of UroLumes in the anterior urethra and in an irradiated urethra led to adverse reactions such as stent separation, poor epithelialization, hyperplastic tissue growth, encrustation, or obstruction necessitating removal. PMID- 9726411 TI - Surgeon's workshop: a simple method of antegrade collagen placement for postprostatectomy incontinence. AB - We describe a modified method for antegrade placement of periurethral collagen in incontinent men. In this technique, a flexible cystoscope is used to guide the placement of a specially designed needle into the bladder neck and external sphincter region via an antegrade transvesical approach. The 19-gauge, 28-cm long needle is passed suprapubically without a sheath. The procedure may be performed with local anesthesia and intravenous sedation only and takes less than 30 minutes. PMID- 9726412 TI - Discrimination between native and intentionally misfolded conformations of proteins: ES/IS, a new method for calculating conformational free energy that uses both dynamics simulations with an explicit solvent and an implicit solvent continuum model. AB - A new method for calculating the total conformational free energy of proteins in water solvent is presented. The method consists of a relatively brief simulation by molecular dynamics with explicit solvent (ES) molecules to produce a set of microstates of the macroscopic conformation. Conformational energy and entropy are obtained from the simulation, the latter in the quasi-harmonic approximation by analysis of the covariance matrix. The implicit solvent (IS) dielectric continuum model is used to calculate the average solvation free energy as the sum of the free energies of creating the solute-size hydrophobic cavity, of the van der Waals solute-solvent interactions, and of the polarization of water solvent by the solute's charges. The reliability of the solvation free energy depends on a number of factors: the details of arrangement of the protein's charges, especially those near the surface; the definition of the molecular surface; and the method chosen for solving the Poisson equation. Molecular dynamics simulation in explicit solvent relaxes the protein's conformation and allows polar surface groups to assume conformations compatible with interaction with solvent, while averaging of internal energy and solvation free energy tend to enhance the precision. Two recently developed methods--SIMS, for calculation of a smooth invariant molecular surface, and FAMBE, for solution of the Poisson equation via a fast adaptive multigrid boundary element--have been employed. The SIMS and FAMBE programs scale linearly with the number of atoms. SIMS is superior to Connolly's MS (molecular surface) program: it is faster, more accurate, and more stable, and it smooths singularities of the molecular surface. Solvation free energies calculated with these two programs do not depend on molecular position or orientation and are stable along a molecular dynamics trajectory. We have applied this method to calculate the conformational free energy of native and intentionally misfolded globular conformations of proteins (the EMBL set of deliberately misfolded proteins) and have obtained good discrimination in favor of the native conformations in all instances. PMID- 9726413 TI - Recognition of single-stranded DNA by nuclease P1: high resolution crystal structures of complexes with substrate analogs. AB - The reaction mechanism of nuclease P1 from Penicillium citrinum has been investigated using single-stranded dithiophosphorylated di-, tetra-, and hexanucleotides as substrate analogs. The complexes crystallize in tetragonal and orthorhombic space groups and have been solved by molecular replacement. The high resolution structures give a clear picture of base recognition by P1 nuclease at its two nucleotide-binding sites, especially the 1.8 A structure of a P1 tetranucleotide complex which can be considered a P1-product complex. The observed binding modes are in agreement with a catalytic mechanism where the two closely spaced zinc ions activate the attacking water while the third, more exposed zinc ion stabilizes the leaving 03' oxyanion. Stacking as well as hydrogen bonding interactions with the base 5' to the cleaved phosphodiester bond are important elements of substrate binding and recognition. Modelling of a productive P1-substrate complex based on the solved structures suggests steric hindrance as the likely reason for the resistance of Rp-phosphorothioates and phosphorodithioates. Differences with the highly homologous nuclease S1 from Aspergillus oryzae are discussed. PMID- 9726414 TI - Statistical mechanics of protein folding by exhaustive enumeration. AB - It is hard to construct theories for the folding of globular proteins because they are large and complicated molecules having enormous numbers of nonnative conformations and having native states that are complicated to describe. Statistical mechanical theories of protein folding are constructed around major simplifying assumptions about the energy as a function of conformation and/or simplifications of the representation of the polypeptide chain, such as one point per residue on a cubic lattice. It is not clear how the results of these theories are affected by their various simplifications. Here we take a very different simplification approach where the chain is accurately represented and the energy of each conformation is calculated by a not unreasonable empirical function. However, the set of amino acid sequences and allowed conformations is so restricted that it becomes computationally feasible to examine them all. Hence we are able to calculate melting curves for thermal denaturation as well as the detailed kinetic pathway of refolding. Such calculations are based on a novel representation of the conformations as points in an abstract 12-dimensional Euclidean conformation space. Fast folding sequences have relatively high melting temperatures, native structures with relatively low energies, small kinetic barriers between local minima, and relatively many conformations in the global energy minimum's watershed. In contrast to other folding theories, these models show no necessary relationship between fast folding and an overall funnel shape to the energy surface, or a large energy gap between the native and the lowest nonnative structure, or the depth of the native energy minimum compared to the roughness of the energy landscape. PMID- 9726415 TI - Can one predict protein stability? An attempt to do so for residue 133 of T4 lysozyme using a combination of free energy derivatives, PROFEC, and free energy perturbation methods. AB - Free energy derivatives, pictorial representation of free energy changes (PROFEC) and free energy perturbation methods were employed to suggest the modifications that may improve the stability of a mutant T4 lysozyme with a S-2-amino-3 cyclopentylpropanoic acid residue (Cpe) at position 133. The free energy derivatives and PROFEC methods were used to locate promising sites where modifications may be introduced. The effects of several candidate modifications on the enzyme's stability were analyzed by the free energy perturbation method. We found that this scheme is able to effectively suggest modifications that may increase the enzyme's stability. The modifications investigated are the introduction of a methyl, a tert-butyl or a trifluoromethyl group at the Cepsilon2 position and a cyclopropyl group between the Cdelta2 and Cepsilon2 position on the cyclopentyl ring. The stereochemistry of the introduced groups (in the alpha or beta configurations) was studied. Our calculations predict that the introduction of a methyl group in the alpha configuration or a cyclopropyl group in the beta configuration will increase the stability of the enzyme; the introduction of the two groups in the other configurations and the other modifications will decrease the stability of the enzyme. The results indicate that packing interactions can strongly influence the stability of the enzyme. PMID- 9726416 TI - Variability of the canonical loop conformations in serine proteinases inhibitors and other proteins. AB - Canonical loops of protein inhibitors of serine proteinases occur in proteins having completely different folds. In this article, conformations of the loops have been analyzed for inhibitors belonging to 10 structurally different families. Using deviation in Calpha-Calpha distances as a criterion for loop similarity, we found that the P3-P3' segment defines most properly the length of the loop. When conformational differences among loops of individual inhibitors were compared using root mean square deviation (rmsd) in atomic coordinates for all main chain atoms (deltar method) and rmsd operating in main chain torsion angles (deltat method), differences of up to 2.1 A and 72.3 degrees, respectively, were observed. Such large values indicate significant conformational differences among individual loops. Nevertheless, the overall geometry of the inhibitor-proteinase interaction is very well preserved, as judged from the similarity of Calpha-Calpha distances between Calpha of catalytic Ser and Calpha of P3-P3' residues in various enzyme-inhibitor complexes. The mode of interaction is very well preserved both in the chymotrypsin and subtilisin families, as distances calculated for subtilisin-inhibitor complexes are almost always within the range of those for chymotrypsin-inhibitor complexes. Complex formation leads to conformational changes of up to 160 degrees for chi1 angle. Side chains of residue P1 and P2' adopt in different complexes a similar orientation (chi1 angle = -60 degrees and -180 degrees, respectively). To check whether the canonical conformation can be found among non-proteinase-inhibitor Brookhaven Protein Data Bank structures, two selection criteria--the allowed main chain dihedral angles and Calpha-Calpha distances for the P3-P3' segment--were applied to all these structures. This procedure detected 132 unique hexapeptide segments in 121 structurally and functionally unrelated proteins. Partial preferences for certain amino acids occurring at particular positions in these hexapeptides could be noted. Further restriction of this set to hexapeptides with a highly exposed P1 residue side chain resulted in 40 segments. The possibility of complexes formation between these segments and serine proteinases was ruled out in molecular modeling due to steric clashes. Several structural features that determine the canonical conformation of the loop both in inhibitors and in other proteins can be distinguished. They include main chain hydrogen bonds both within the P3-P3' segment and with the scaffold region, P3-P4 and P3'-P4' hydrophobic interactions, and finally either hydrophobic or polar interactions involving the P1' residue. PMID- 9726417 TI - Assembly of protein structure from sparse experimental data: an efficient Monte Carlo model. AB - A new, efficient method for the assembly of protein tertiary structure from known, loosely encoded secondary structure restraints and sparse information about exact side chain contacts is proposed and evaluated. The method is based on a new, very simple method for the reduced modeling of protein structure and dynamics, where the protein is described as a lattice chain connecting side chain centers of mass rather than Calphas. The model has implicit built-in multibody correlations that simulate short- and long-range packing preferences, hydrogen bonding cooperativity and a mean force potential describing hydrophobic interactions. Due to the simplicity of the protein representation and definition of the model force field, the Monte Carlo algorithm is at least an order of magnitude faster than previously published Monte Carlo algorithms for structure assembly. In contrast to existing algorithms, the new method requires a smaller number of tertiary restraints for successful fold assembly; on average, one for every seven residues as compared to one for every four residues. For example, for smaller proteins such as the B domain of protein G, the resulting structures have a coordinate root mean square deviation (cRMSD), which is about 3 A from the experimental structure; for myoglobin, structures whose backbone cRMSD is 4.3 A are produced, and for a 247-residue TIM barrel, the cRMSD of the resulting folds is about 6 A. As would be expected, increasing the number of tertiary restraints improves the accuracy of the assembled structures. The reliability and robustness of the new method should enable its routine application in model building protocols based on various (very sparse) experimentally derived structural restraints. PMID- 9726418 TI - Multiple conformational states of a new hematopoietic cytokine (megakaryocyte growth and development factor): pH- and urea-induced denaturation. AB - The effect of pH and urea on the conformation of recombinant human megakaryocyte growth and development factor (rHuMGDF) was determined by circular dichroism, intrinsic fluorescence spectroscopy, and equilibrium ultracentrifugation. The conformation of rHuMGDF was dependent on pH and urea concentration. Multiple folding forms were evidenced by multiple pH-induced transitions and urea-induced equilibrium transitions that deviated from a simple two-state process. In neutral to alkaline pH, rHuMGDF exists as a monomer, but an acid-induced conformational state self-associates to form a soluble aggregate. A folding intermediate(s) was observed with a more stable secondary structure than tertiary structure and was dependent on the pH of the urea-induced denaturation. The differences in the stabilities of the folding states were most distinct in the pH range of 4.5 to 6.5. The presence of intermediates in the folding pathway of rHuMGDF are similar to findings of previous studies of related growth factors that share a common three-dimensional structure. PMID- 9726419 TI - Structural basis for specificity of papain-like cysteine protease proregions toward their cognate enzymes. AB - Synthetic peptides corresponding to the proregions of papain-like cysteine proteases have been shown to be good and selective inhibitors of their parental enzymes. The molecular basis for their selectivity, quite remarkable in some cases, is not fully understood. The recent determination of the crystal structures of three distinct papain-like cysteine protease zymogens allows detailed structural comparisons to be made. The reasons for the specificity shown by each proregion toward its cognate enzyme are explained in terms of the three dimensional structure of the proregion and the interface between the mature enzyme and the proregion. These comparisons reveal that insertion and substitution of amino acids within the proregion cause major rearrangement of sidechains on the enzyme/proregion interface, allowing detailed surface and charge recognition. PMID- 9726420 TI - Camel single-domain antibody inhibits enzyme by mimicking carbohydrate substrate. AB - Whereas antibodies have demonstrated the ability to mimic various compounds, classic heavy/light-chain antibodies may be limited in their applications. First, they tend not to bind enzyme active site clefts. Second, their size and complexity present problems in identifying key elements for binding and in using these elements to produce clinically valuable compounds. We have previously shown how cAb-Lys3, a single variable domain fragment derived from a lysozyme-specific camel antibody naturally lacking light chains, overcomes the first limitation to become the first antibody structure observed penetrating an enzyme active site. We now demonstrate how cAb-Lys3 mimics the oligosaccharide substrate functionally (inhibition constant for lysozyme, 50 nM) and structurally (lysozyme buried surface areas, hydrogen bond partners, and hydrophobic contacts are similar to those seen in sugar-complexed structures). Most striking is the mimicry by the antibody complementary determining region 3 (CDR3) loop, especially Ala104, which mimics the subsite C sugar 2-acetamido group; this group has previously been identified as a key feature in binding lysozyme. Comparative simplicity, high affinity and specificity, potential to reach and interact with active sites, and ability to mimic substrate suggest that camel heavy-chain antibodies present advantages over classic antibodies in the design, production, and application of clinically valuable compounds. PMID- 9726421 TI - Reactivation of the totally inactive pancreatic lipase RP1 by structure-predicted point mutations. AB - Both classical pancreatic lipase (DPL) and pancreatic lipase-related protein 1 (DPLRP1) have been found to be secreted by dog exocrine pancreas. These two proteins were purified to homogeneity from canine pancreatic juice and no significant catalytic activity was observed with dog PLRP1 on any of the substrates tested: di- and tri-glycerides, phospholipids, etc. DPLRP1 was crystallized and its structure solved by molecular replacement and refined at a resolution of 2.10 A. Its structure is similar to that of the classical PL structures in the absence of any inhibitors or micelles. The lid domain that controls the access to the active site was found to have a closed conformation. An amino-acid substitution (Ala 178 Val) in the DPLRP1 may result in a steric clash with one of the acyl chains observed in the structures of a C11 alkyl phosphonate inhibitor, a transition state analogue, bound to the classical PL. This substitution was suspected of being responsible for the absence of DPLRP1 activity. The presence of Val and Ala residues in positions 178 and 180, respectively, are characteristic of all the known PLRP1, whereas Ala and Pro residues are always present in the same positions in all the other members of the PL gene family. Introducing the double mutation Val 178 Ala and Ala 180 Pro into the human pancreatic RP1 (HPLRP1) gene yielded a well expressed and folded enzyme in insect cells. This enzyme is kinetically active on triglycerides. Our findings on DPLRP1 and HPLRP1 are therefore likely to apply to all the RP1 lipases. PMID- 9726423 TI - Platelet-derived growth factor induces chemotaxis of neuroepithelial stem cells. AB - The ability of differentiating cells to migrate within the developing central nervous system (CNS) depends on extrinsic guidance signals, some of which are growth factors. In this study we have investigated the chemotactic response of cultured stem cells from the embryonic rat cortex to platelet-derived growth factor (PDGF). Nestin-positive stem cells from the developing CNS can be maintained and expanded in vitro under serum-free conditions in the presence of basic fibroblast growth factor (bFGF). Northern blot analysis of PDGF receptor expression revealed both alpha- and beta-receptors on bFGF-treated neural stem cells. Both PDGF-AA and PDGF-BB readily induced directed migration of cultured neuroepithelial cells as measured in a microchemotaxis assay. Blocking of the migratory response was achieved by incubation with PDGF isoform-specific antibodies. More than 90% of the migrating cells were nestin-positive and incorporation of BrdU was also seen suggesting the cells to be immature and not yet committed to a specific cell lineage. These findings suggest a role for PDGF in cell migration in the developing cortex. PMID- 9726422 TI - Potassium currents and excitability in second-order auditory and vestibular neurons. AB - Potassium channels are involved in the control of neuronal excitability by fixing the membrane potential, shaping the action potential, and setting firing rates. Recently, attention has been focused on identifying the factors influencing excitability in second-order auditory and vestibular neurons. Located in the brainstem, second-order auditory and vestibular neurons are sites for convergence of inputs from first-order auditory or vestibular ganglionic cells with other sensory systems and also motor areas. Typically, second-order auditory neurons exhibit two distinct firing patterns in response to depolarization: tonic, with a repetitive firing of action potentials, and phasic, characterized by only one or a few action potentials. In contrast, all mature vestibular second-order neurons fire tonically on depolarization. Already, certain fundamental roles have emerged for potassium currents in these neurons. In mature auditory and vestibular neurons, I(K), the delayed rectifier, is required for the fast repolarization of action potentials. In tonically firing auditory neurons, I(A), the transient outward rectifier, defines the discharge pattern. I(DS), a delayed rectifier-like current distinguished by its low threshold of activation, is found in phasically firing auditory and some developing vestibular neurons where it limits firing to one or a few spikes, and also may contribute to forming short-duration excitatory postsynaptic potential (EPSPs). Also, I(DS) sets the threshold for action potential generation rather high, which may prevent spontaneous discharge in phasically firing cells. During development, there is a gradual acquisition and loss of some potassium conductances, suggesting developmental regulation. As there are similarities in membrane properties of second-order auditory and vestibular neurons, investigations on firing pattern and its underlying mechanisms in one system should help to uncover fundamental properties of the other. PMID- 9726424 TI - Decreased expression of the metabotropic glutamate receptor-4 gene is associated with neuronal apoptosis. AB - Cultured cerebellar granule neurons die by apoptosis when switched from medium containing elevated potassium (K+) and serum to serum-free medium containing low K+. Although cell death begins at about 16 hr, commitment to death occurs within 6 hr after the lowering of K+. We have used this paradigm to examine the role of metabotropic glutamate receptor (mGluR) genes in the regulation of neuronal survival. We find that the expression of one of the mGluR genes, the type-4 gene, is associated with increased neuronal survival. Lowering of K+ leads to an 80% decrease in mGluR-4 mRNA expression within 6 hr. Downregulation of mGluR-4 messenger RNA (mRNA) does not occur if low K+-induced death is prevented by treatment with insulin-like growth factor I or adenosine 3',5'-cyclic monophosphate. If transcription is inhibited by actinomycin D, the difference in mGluR4 mRNA expression between cells switched to high-K+ medium and those switched to low-K+ medium is dramatically reduced, suggesting that decreased mGluR-4 gene transcription rather than increased mRNA breakdown is mainly responsible for the apoptosis-associated decrease in mGluR4 levels. Blockade of transcription also reduces mGluR4 mRNA expression in healthy neurons by more than 50% within 4 hr, suggesting that the mGluR4 mRNA has a relatively short half life. In pharmacological experiments, we observe that the specific group III mGluR agonists such as L-amino-4-phosphobutyric acid and O-phospho-L-serine inhibit low K+-induced apoptosis. On the other hand, a selective mGluR4 antagonist, (RS)-alpha-cyclopropyl-4-phosphono-phenylglycine, induces apoptosis even in the presence of elevated K+. These results indicate that elevated mGluR4 expression or the activation of this receptor promotes survival and that an inhibition of such survival mechanisms contributes to apoptosis. PMID- 9726425 TI - Accelerated demyelination of peripheral nerves in mice deficient in connexin 32 and protein zero. AB - Mutant mice that lack either protein zero (P0) or connexin 32 (Cx32) were generated previously to investigate the function of these myelin proteins in peripheral nerves and to assess the value of these mice as animal models for hereditary human peripheral neuropathies. Mice that are completely devoid of P0 expression (P0(+/0)) show a complex phenotype that is characterized by hypomyelination, compromised myelin compaction, and degeneration of myelin and axons early in life. In contrast, young mouse mutants that have retained one wild type allele of the P0 gene (P0(+/0)) reveal morphologically normal myelin but start to develop signs of demyelination and remyelination at 4 months of age. A similar late-onset myelin deficiency was observed in Cx32-deficient mice (Cx32(0/0)). We have now generated mice deficient for Cx32 and P0. In animals that lack both proteins (Cx32(0/0)/P0(0/0), the phenotype is morphologically identical to mice that solely lack P0. Animals that lack Cx32 and carry one functional P0 allele (Cx32(0/0/P0(+/0)) revealed demyelination and remyelination as evidenced by thin myelin and Schwann cell onion bulb formation already at the age of 4 weeks, a time point when no pathology was observed in the single mutants. These morphological deficits were also more prominent in 4-month-old Cx32(0/0)/P0(+/0)animals compared to the single mutants. Our data support the view that Cx32 and P0 are crucial molecules for the maintenance of myelin. Furthermore, the function of Cx32 in the peripheral nervous system appears to be largely dispensable when myelin compaction is impaired. PMID- 9726426 TI - Antibodies from patients with Parkinson's disease react with protein modified by dopamine oxidation. AB - To determine whether specific antibodies are present in PD, we used an enzyme linked immunosorbant assay (ELISA) that identifies increased immunoglobulin (IgG) levels towards a synthetic substrate prepared by incubating ovalbumin with dopamine and copper sulfate. Altered absorption spectrum and specific chemical detection demonstrated quinone modification of the ovalbumin. This modified protein was demonstrated to react with serial dilutions of PD sera. A threshold dilution of 1:500 was subsequently used to screen sera from patients with PD (n = 21), amyotrophic lateral sclerosis (n = 7), Alzheimer's disease (n = 7) and other neurological disease controls (n = 7). The assay produced a positive result in 7/21 PD patients and 0/21 disease controls (P < 0.02, Kruskal-Wallis test). Further testing of sera from untreated PD patients (n = 6) identified one positive sample. Thus, a subset of Parkinson's disease (PD) patients has immunoglobulin (IgG) to ovalbumin modified by dopamine oxidation. The presence of antibody reactivity to quinone-modified proteins could contribute to or amplify the inflammatory response in PD. PMID- 9726427 TI - Potentiating interactions between morphogenetic protein and neurotrophic factors in developing neurons. AB - mRNA for bone morphogenetic protein receptor type II (BMPR-II) was mapped to different neurons in peripheral ganglia and spinal cord of the chicken embryo. The expression of this serine/threonine kinase receptor partially overlaps with that of tyrosine kinase receptors Trk and Ret. Biological activities of osteogenic protein-1 (OP-1), a documented ligand for BMPR-II, were tested in explanted embryonic chicken ganglia and dissociated ganglionic neurons. OP-1 had only a limited stimulatory effect on neuronal survival. However, OP-1 combined with either neurotrophin-3 (NT-3, a relative of nerve growth factor) or glial cell line-derived neurotrophic factor (GDNF) potentiated neuronal survival three- to fourfold. We also show that OP-1 strongly potentiates nerve fiber outgrowth from ganglia stimulated with NT-3 or GDNF. Signaling by BMPR-II in neurons may potentiate the tyrosine kinase pathway activated by NT-3 and GDNF. The data suggest that morphogenetic proteins may modulate neurotrophic activities during neuronal development and plasticity. PMID- 9726428 TI - Neuronal differentiation of chromaffin cells in vitro, induced by extremely low frequency magnetic fields or nerve growth factor: a histological and ultrastructural comparative study. AB - The application of nerve growth factor (NGF) to primary adrenal medulla chromaffin cell cultures induces phenotypic changes characterized mainly by the presence of neurites. A similar effect has been seen when these cells are stimulated by extremely low frequency magnetic fields (ELFMF). In this study, newborn rat chromaffin cells were cultured and subjected to NGF or ELFMF in order to compare their histological and ultrastructural characteristics. Cells cultured in the presence of NGF developed cytoplasmic projections and their distal ends showed growth cones as well as filopodia. With scanning and transmission electron microscopy, an increased submembranous electron density was observed in the nuclei of cells as well as irregular, wavy neuritic projections with a moderate number of varicosities, as well as the prevalence of intermediate filaments among the cytoskeleton components. Cells stimulated with ELFMF presented straighter neuritic extensions with a greater number of varicosities. With the transmission electron microscope, numerous neurotubules were observed, both in the cell soma and in their neuritic extensions. In both groups, growth cones were clearly identified by their ultrastructural characteristics. The differences seen in the cytoskeleton of cells stimulated with NGF or ELFMF suggest differential stimulation mechanisms possibly determining the biochemical, electrophysiological, and morphological characteristics in both types of cell cultures. PMID- 9726429 TI - Strong nuclear factor-kappaB-DNA binding parallels cyclooxygenase-2 gene transcription in aging and in sporadic Alzheimer's disease superior temporal lobe neocortex. AB - Cyclooxygenase-2 (COX-2; EC 1.14.99.1) RNA message abundance in 25 control and Consortium to Establish a Registry for Alzheimer's Disease (CERAD)-confirmed sporadic Alzheimer's disease (AD) brains is remarkably heterogeneous when compared with 55 other AD brain RNA message levels that were previously characterized (Lukiw and Bazan: J Neurosci Res 50:937-945, 1997). Examination of nuclear protein extracts (NPXTs) that were derived from control and AD-affected brain neocortical nuclei (n = 20; age range, 60-82 years; postmortem interval, 0.5-6.5 hours) by using gel shift, gel supershift, and cold oligonucleotide competition assay revealed a highly significant relationship between the extent of inflammatory transcription factor, nuclear factor (NF)-kappaB: DNA binding and the abundance of the COX-2 RNA signal (P < 0.0001; analysis of variance). No strong correlation with AP-1-DNA binding was noted (P > 0.045). These data are the first linking inflammation-related transcription factor NF-KB-DNA binding to up-regulation of transcription from a key inflammatory gene, COX-2, in both normally aging brain and in AD-affected neocortex. Systematic deletion of NF-KB DNA binding sites in human COX-2 promoter constructs attenuates COX-2 transcriptional induction by mediators of inflammation. Strong NF-kappaB-DNA binding has been reported previously to temporally precede COX-2 gene transcription in human epithelial (A549), hamster B-cell (HIT-T15), human endothelial (HUVEC), human lymphoblast (IM9), human fibroblast (IMR90), rat glioma/mouse neuroblastoma (NG108-15), human keratinocyte (NHEK), mouse fibroblast (NIH 3T3), rat neuroblastoma (SH-SY5Y) cell lines and in mouse and rat brain hippocampus, indicating a highly conserved inflammatory signaling pathway that is common to diverse species and cell types. The mouse, rat, and human COX-2 immediate promoters, despite 7.5 x 10(7) years of DNA sequence divergence, each retain multiple recognition sites specific for NF-kappaB-DNA binding. These data suggest that basic gene induction mechanisms, which have been conserved over long periods of evolution, that increase NF-kappaB-DNA binds ing may be fundamental in driving transcription from inflammation-related genes, such as COX-2, that operate in stressed tissues, in normally aging cell lines, and in neurodegenerative disorders that include AD brain. PMID- 9726430 TI - Enhanced glial cell line-derived neurotrophic factor mRNA expression upon (-) deprenyl and melatonin treatments. AB - Glial cell line-derived neurotrophic factor (GDNF) has been shown to be a preferentially selective neurotrophic factor for dopamine (DA) neurons. In the present study, we have examined the distribution of GDNF mRNA expression in several major DA-containing cell body and terminal areas and the regulation of GDNF mRNA expression upon various pharmacological treatments. Results indicated that there is a relatively higher GDNF mRNA level in neurons of the nigrostriatal and mesolimbic dopaminergic pathways. Upon chronic 1-methyl-4-phenyl1,2,3,6 tetrahydropyridine (MPTP) treatment (30 mg/ kg, i.p., for 7 days), DA level was decreased, whereas GDNF mRNA expression was increased in the striatum, suggesting that more GDNF is synthesized and expressed to cope with the neurotoxin insult. Furthermore, among several DA neuron protective and/or therapeutic agents examined, both intrastriatal injections of (-)-deprenyl (1.25 microg and 2.5 microg) and melatonin (30 microg, 60 microg, and 120 microg) significantly enhanced GDNF mRNA expression in the striatum, whereas the same concentrations of (-)-deprenyl did not affect monoamine oxidase B (MAOB) activity, although it increased glutathione peroxidase (GPx) and/or superoxide dismutase (SOD) activities. Similarly, the same concentrations of melatonin did not alter SOD or GPx activities, except that the highest dose of melatonin (120 microg) increased lipid peroxidation in the striatum. Conversely, GM1 ganglioside injection (45 microg) lacked of an effect on GDNF mRNA expression. Together, these results suggest that both (-)-deprenyl and melatonin up-regulate GDNF gene expression at threshold doses lower than that needed for altering MAOB activity and/or the antioxidant enzyme systems, respectively. These results provide new information on the neuroprotective and therapeutic mechanisms of (-)-deprenyl and melatonin on DA neurons. PMID- 9726431 TI - Binding of cholera toxin B subunit: a surface marker for murine microglia but not oligodendrocytes or astrocytes. AB - GM1 ganglioside is a receptor for the B subunit of cholera toxin. In lymphocytes, B subunit elicits an influx of extracellular Ca++ (Dixon et al., 1987). To investigate this signaling pathway in glia, we assessed the presence of GM1 ganglioside on the surface of cultured murine central nervous system (CNS) glia by binding of fluorescein-labeled B subunit. B subunit binding was compared to binding of peanut agglutinin, wheat germ agglutinin, and Bandeiraea (Griffonia) simplicifolia lectin (BSL)I, a microglial marker. Antibodies to glial fibrillary acidic protein, A007/O4 antigens, and galactocerebroside were used to identify astrocytes, immature oligodendrocytes (OLs) and mature OLs, respectively. Binding patterns differed based on cell type and developmental stage. Wheat germ and peanut agglutinins bound to the surface of microglia, astrocytes, and immature OLs; neither lectin bound to any significant extent to the surface of membrane sheets of mature OLs, although wheat germ agglutinin was rapidly endocytosed. Cells identified as microglia by BSL I binding and morphology were the only cells to stain brightly on the surface with B subunit. Thus, surface GM1 ganglioside appears to be a highly enriched marker for microglia in these mixed glial cultures. The effects of B subunit on intracellular Ca++ were examined by laser cytometry in glial cultures loaded with Indo-1. No Ca++ responses were observed in microglia. Mature OLs were examined for Ca++ responses to B subunit before and after surface levels of GM1 ganglioside were increased by incubation with exogenous GM1 ganglioside. Again, no Ca++ responses were observed. Thus, cultured microglia and mature OLs do not have the GM1-mediated signal transduction pathway seen in lymphocytes. However, the presence of GM1 ganglioside on microglia may play a role in giving rise to antibodies to this glycolipid in some CNS inflammatory diseases. PMID- 9726432 TI - Uric acid protects neurons against excitotoxic and metabolic insults in cell culture, and against focal ischemic brain injury in vivo. AB - Uric acid is a well-known natural antioxidant present in fluids and tissues throughout the body. Oxyradical production and cellular calcium overload are believed to contribute to the damage and death of neurons that occurs following cerebral ischemia in victims of stroke. We now report that uric acid protects cultured rat hippocampal neurons against cell death induced by insults relevant to the pathogenesis of cerebral ischemia, including exposure to the excitatory amino acid glutamate and the metabolic poison cyanide. Confocal laser scanning microscope analyses showed that uric acid suppresses the accumulation of reactive oxygen species (hydrogen peroxide and peroxynitrite), and lipid peroxidation, associated with each insult. Mitochondrial function was compromised by the excitotoxic and metabolic insults, and was preserved in neurons treated with uric acid. Delayed elevations of intracellular free calcium levels induced by glutamate and cyanide were significantly attenuated in neurons treated with uric acid. These data demonstrate a neuroprotective action of uric acid that involves suppression of oxyradical accumulation, stabilization of calcium homeostasis, and preservation of mitochondrial function. Administration of uric acid to adult rats either 24 hr prior to middle cerebral artery occlusion (62.5 mg uric acid/kg, intraperitoneally) or 1 hr following reperfusion (16 mg uric acid/kg, intravenously) resulted in a highly significant reduction in ischemic damage to cerebral cortex and striatum, and improved behavioral outcome. These findings support a central role for oxyradicals in excitotoxic and ischemic neuronal injury, and suggest a potential therapeutic use for uric acid in ischemic stroke and related neurodegenerative conditions. PMID- 9726433 TI - Ca2+ changes and 86Rb efflux activated by hyposmolarity in cerebellar granule neurons. AB - Hyposmotic swelling increased 86Rb release in cultured cerebellar granule neurons (1 day in vitro [DIV]) with a magnitude related to the change in osmolarity. 86Rb release was partially blocked by quinidine, Ba2+, and Cs+ but not by TEA, 4-AP, or Gd3+. 86Rb efflux decreased in Cl(-)-depleted cells or cells treated with DDF or DIDS, suggesting an interconnection between Cl- and K+ fluxes. Swelling induced a substantial increase in [Ca2+]i to which both external and internal sources contribute. However, 86Rb efflux was independent of [Ca2+]0, unaffected by depleting the endoplasmic reticulum (ER) by ionomycin or thapsigargin and insensitive to charybdotoxin, iberiotoxin, and apamin. Swelling-activated 86Rb efflux in differentiated granule neurons after 8 DIV, which express Ca2+ sensitive K+ channels, was not different from that in 1 DIV neurons, nor in time course, net release, Ca2+-dependence, or pharmacological sensitivity. We conclude that the swelling-activated K+ efflux in cerebellar granule neurons is not mediated by Ca2+-sensitive large conductance K+ channels (BK) as in many cell types but resembles that in lymphocytes where it is possibly carried by voltage gated K+ channels. PMID- 9726434 TI - Studies of IFN-induced transcriptional activation uncover the Jak-Stat pathway. AB - Sharing the Milstein Award with George R. Stark and Ian M. Kerr in the fall of 1997 brought this invitation to record personal reflections on our experiments concerning the mechanisms of action of interferon. Our work and that of the Kerr and Stark laboratories uncovered the Jak-Stat pathway through which signals from cell surface receptors reach genes in the cell nucleus. This review concentrates on that to which I can speak most reliably, that is, experiments done by my young colleagues at Rockefeller University. PMID- 9726435 TI - Proinflammatory cytokines lowering erythropoietin production. AB - The plasma level of erythropoietin (Epo) in anemic patients suffering from inflammation is often low in relation to the blood hemoglobin concentration. Various proinflammatory cytokines have been tested for their action on the synthesis of Epo. Interleukin 1 (IL-1) and tumor necrosis factor-alpha(TNF-alpha) suppress Epo gene expression in isolated perfused rat kidneys and in human hepatoma cell cultures. IL-6 inhibits in the kidney, and conflicting results have been reported for its effect on Epo synthesis in hepatic cells. Several other cytokines tested were without effect. Thus, mainly IL-1 and TNF-alpha seem to be responsible for the defect in Epo production in severe systemic and renal inflammatory diseases. PMID- 9726436 TI - Recombinant IFN-gamma treatment of a patient with hyperimmunoglobulin E syndrome triggered autoimmune thrombocytopenia. AB - We report a pediatric patient with hyperimmunoglobulin E syndrome (HIES) treated with recombinant IFN-gamma (rIFN-gamma) for 2 1/4 years who developed autoimmune thrombocytopenia and was positive for serum antiplatelet antibody and antinuclear antibody (ANA). She was then treated with i.v. methylprednisolone pulse therapy followed by oral immunosuppressive drugs. With this therapy, her platelet count increased and was maintained within the normal range for more than a year. We retrospectively examined her sera stored at -40 degrees C for ANA and found that the ANA level was increased from 1:40 to 1:640 with the rIFN-y therapy. Therefore, we believe that, in this case, rIFN-y treatment may have played a crucial role in triggering autoimmune thrombocytopenia. Furthermore, this case demonstrates that caution must be observed in administering rIFN-gamma to genetically predisposed patients. PMID- 9726437 TI - Biomodulation with sequential intravenous IFN-alpha2b and 5-fluorouracil as second-line treatment in patients with advanced colorectal cancer. AB - A phase II trial was carried out by the Grupo Oncologico Cooperativo del Sur (G.O.C.S.) to assess the efficacy and toxicity of a biochemical modulation of 5 fluorouracil (5-FU) by i.v. pretreatment with interferon (IFN)-alpha2b in patients with advanced colorectal carcinoma refractory to previous therapy with 5 FU modulated by methotrexate (MTX) or leucovorin (LV) or both. Between January 1993 and October 1995, 34 patients were entered on the study. The treatment was IFN-alpha2b 5 x 10(6)/m2 IU in a 1-h i.v. infusion, followed immediately by 5-FU 600 mg/m2 i.v. bolus injection. Courses were repeated weekly until observation of progressive disease or severe toxicity. One patient could not be assessed for response. Objective regression was observed in 2 of 33 patients (6%, 95% confidence interval, 0%-14%). No patient achieved a complete response. Two patients had partial responses (6%). No change was recorded in 14 patients (41%), and progressive disease occurred in 17 (52%). The median time to treatment failure was 3 months, and the median survival was 5 months. Toxicity was within acceptable limits. The main side effects were mucositis and diarrhea. Four episodes of grade 2 stomatitis were observed, causing dosage modifications. The most frequent toxic effects attributable to IFN-alpha2b were mild fatigue and fever. In conclusion, second-line therapy with i.v. IFN-alpha2b preceding 5-FU has shown an interesting profile of activity in a patient population with clearly unfavorable characteristics. From this perspective, further appropriately designed studies are needed to identify the greatest potential of IFN-alpha2b as a modulator of 5-FU. PMID- 9726438 TI - IFN-alpha2b suppresses the fibrogenic effects of insulin-like growth factor-1 in dermal fibroblasts. AB - The interferon (IFN) proteins, including IFN-alpha2b have been used as antifibrogenic factors to modulate the expression of extracellular matrix (ECM) proteins associated with fibroproliferative disorders in skin. This study was conducted to determine if IFN-alpha2b can counteract the fibrogenic effects of insulin-like growth factor-1 (IGF-1), which is present in large quantity in fibrotic dermis. Human dermal fibroblasts were established in culture and treated with either vehicle (control), 2000 U/ml IFN-alpha2b alone, 100 ng/ml IGF-1 alone, or both IFN-alpha2b and IGF-1. The results showed that treatment with IFN alpha2b inhibited the proliferation of dermal fibroblasts, reduced the steady state levels of type I procollagen mRNA in the cells, and reduced the production of collagen as measured by hydroxyproline in conditioned medium. However, this treatment also increased levels of collagenase mRNA in the cells and collagenase activity in the medium. Cells treated with IGF-1 showed increased proliferation and collagen production and decreased collagenase. Cells treated with both IFN alpha2b and IGF-1 exhibited a 44% reduction in hydroxyproline production (p < 0.05) and a 363% increase in collagenase activity over cells treated with IGF-1 alone (p < 0.01). These results indicate that when IGF-1 and IFN-alpha2b are used individually, they function as fibrogenic and antifibrogenic factors for dermal fibroblasts, respectively, and that fibrogenic effects of IGF-1 on cell proliferation, collagen, and collagenase expression can be counteracted by IFN alpha2b. These findings support the potential use of IFN-alpha2b as a therapeutic agent for treatment of fibroproliferative disorders, such as postburn hypertrophic scarring. PMID- 9726439 TI - Granulocyte colony-stimulating factor activates protein kinase A in granulocytic but not monocytic precursors or neutrophils. AB - Granulocyte colony-stimulating factor (G-CSF) regulates survival, proliferation, differentiation, and activation of myeloid cells. G-CSF-R signaling mechanisms other than tyrosine kinase activation have not been documented. We explored the potential involvement of cAMP-dependent protein kinase A (PKA) in G-CSF-R signal transduction. In this report, we provide the first direct evidence of PKA modulation by G-CSF-R. G-CSF treatment of granulocytic precursor cell lines (HL 60, NFS-60, KG-1) resulted in PKA activation, measured by phosphorylation of Kemptide, a peptide substrate. In contrast, the myelomonocytic cell lines (WEHI 3B,U-937) and peripheral blood neutrophils (PMNC) showed a rapid decrease in PKA activity in response to G-CSF. H-89, a specific inhibitor of PKA, blocked G-CSF induced PKA activation in HL-60 cells but did not affect ligand-induced downmodulation of G-CSF-R. Indomethacin, an inhibitor of the cyclooxygenase pathway and prostaglandin synthesis, did not inhibit PKA induction in G-CSF treated HL-60 cells. Our results demonstrate the involvement of PKA in G-CSF-R signal transduction and suggest a lineage-restricted, developmental stage specific regulation of this pathway in myeloid cells. PMID- 9726440 TI - Cytokines produced early in picornavirus infection reflect resistance or susceptibility to disease. AB - Gender bias favoring female resistance to picornavirus disease is not seen in ICR Swiss mice following infection with the MM strain of encephalomyocarditis virus (EMCV) (causing encephalitis and death) as it is with D variant of EMCV (causing diabetes in males). To define this difference, an in vitro virus-infected splenocyte culture system was used to explore virus effects on lymphoid cells. Infected and sham-infected splenocyte cultures, prepared from both genders of mice and infected with either virus variant, were examined for immunoregulatory cytokines in the first 24 h of infection using ELISA or bioassays. Disease resistance was associated with increased levels of interferon-y (IFN-gamma) and undetectable levels of interleukin-10 (IL-10) by 12 h postinfection in splenocytes from ICR Swiss females infected with EMCV-D. Disease susceptibility was associated with high levels of IL-10 at 12 h after infection of spleen cells from ICR Swiss males infected with EMCV-D or from both genders infected with EMCV MM. This information was used to protect susceptible mice against picornavirus disease (either diabetes or death) by giving them an inducer of IFN-alpha/beta, to induce natural killer (NK)-like cells to produce high levels of IFN-gamma and rat monoclonal anti-IL-10 to neutralize the effects of mouse IL-10. PMID- 9726442 TI - Genomic features of human PKR: alternative splicing and a polymorphic CGG repeat in the 5'-untranslated region. AB - The double-stranded RNA-dependent protein kinase (PKR) is a serine/threonine kinase that plays an important role in the antiviral activities of interferon (IFN). To determine the organization and regulation of the PKR locus, lambda phage and bacterial artificial chromosome (BAC) clones containing the human PKR gene were isolated. Characterization of these clones revealed that PKR has 17 exons and 16 introns dispersed in a genomic region of 50 kb. Sequence analysis of the PKR 5'-flanking region identified a canonical IFN-stimulated response element (ISRE), GAAAACGAAACT. Transient transfection of PKR promoter constructs linked to a luciferase reporter gene into human T98G cells indicated that this 5'-flanking region is capable of functioning as a basal promoter that is also inducible by IFN-alpha and IFN-beta but not IFN-gamma. Interestingly, the PKR gene contains a polymorphic CGG trinucleotide repeat in exon 1. In addition, four PKR alleles, varying in repeat number from 7 to 10, were detected in 30 individual chromosomes. The PKR gene undergoes alternative splicing of exon 2, which gives rise to two forms of 5'-untranslated exons of different length. Although the human and murine PKR proteins have high homology, comparison of their gene structures reveals divergence in 5'-flanking regions, suggesting distinct regulation at the genomic level. PMID- 9726441 TI - An IgG3-IL-2 fusion protein recognizing a murine B cell lymphoma exhibits effective tumor imaging and antitumor activity. AB - Antibody (Ab)-based tumor therapeutics use the tumor-binding specificity of the Ab to target Fc functions or associated molecules to the site of the tumor. We have used an Ab-interleukin-2 (IL-2) fusion protein to deliver IL-2 to a murine B cell lymphoma (38C13). This anti-Id IgG3-CH3-IL-2, which recognizes the idiotype present on the surface of the lymphoma has a half-life in mice approximately 17 fold longer than the half-life reported for IL-2. Gamma camera studies showed that anti-Id IgG3-CH3-IL-2 localizes at the site of a subcutaneous tumor in mice. The anti-Id IgG3-CH3-IL-2 also shows enhanced antitumor activity compared with the combination of Ab and IL-2 administered together. However, the mechanism of antitumor activity appears to depend on the dose and the treatment schedule used. A single dose of fusion protein prevented tumor in only 50% of the animals, although all the survivors showed some evidence of immunologic memory. Although multiple doses are more effective in preventing tumor growth (87% survivors), they are ineffective in generating protective immunologic memory. Our results suggest that Ab-IL-2 fusion proteins will be useful in the diagnosis and treatment of human B cell lymphomas and other related malignancies. PMID- 9726443 TI - Coadministration of IFN-gamma enhances antibody responses in chickens. AB - The development of new generation vaccines has focused on the use of natural immunologic adjuvants that are capable of enhancing a protective immune response. The use of cytokines as immunomodulators in livestock animals, particularly poultry, is becoming more feasible with the recent cloning of several cytokine genes and the progression of new delivery technologies, such as live vectors and DNA delivery. Given that chickens are reared under intensive conditions that are conducive to infection by opportunistic pathogens, the primary mechanism for disease control in poultry is early and effective vaccination. However, many poultry vaccines offer only short-term protection or give nonuniform responses within flocks. We have developed a model system with which to measure the adjuvant potential of cytokines in chickens. This involves measuring antibody levels following coadministration of chicken interferon-gamma (Ch-IFN-gamma) with sheep red blood cells (SRBC). Groups of SPF and commercial broiler birds were injected with two different doses of SRBC with and without coadministration of Ch IFN-y. Three weeks later, all birds were boosted with SRBC alone. Sera were collected weekly and anti-SRBC antibody titers (total Ig and IgG) were determined by hemagglutination. Priming Ch-IFN-gamma resulted in enhanced primary and secondary (IgG) antibody responses that persisted at higher levels when compared with birds that received SRBC alone. Second, coadministration of Ch-IFN-y allowed a 10-fold lower dose of antigen to be as effective as a high dose of antigen that was given without Ch-IFN-gamma. Third, treatment with Ch-IFN-y resulted in an increase in the proportion of birds responding to antigen challenge. These results suggest the potential use for Ch-IFN-gamma as a vaccine adjuvant. PMID- 9726445 TI - Cytokine expression in hepatocytes: role of oxidant stress. AB - Inflammatory mediators, including cytokines and chemokines, are associated with the pathology of chronic liver disease. Interleukin-8 (IL-8) in humans and macrophage inflammatory protein-2 (MIP-2) in rodents, both members of the C-X-C family of chemokines, are particularly potent neutrophil attractants and have been implicated in chronic liver diseases. In the liver, cytokine secretion is usually associated with non-parenchymal cells, particularly Kupffer cells. In the present studies, chemokine gene expression and secretion were investigated in hepatocytes treated with various stimulators. Using human Hep G2 cells, it was demonstrated that, in contrast to lipopolysaccharides (LPS), both tumor necrosis factor-alpha (TNF-beta) and H2O2 are potent inducers of IL-8, presumably acting via protein kinase C (PKC)-dependent pathways. MIP-2 expression occurred in freshly isolated rat hepatocytes following treatment with TNF-alpha, LPS, and to a lesser degree, H2O2. Both IL-8 and MIP-2 secretion were inhibited, although to varying degrees, by such antioxidants as TMTU, DMSO, catalase, and N acetylcysteine. Furthermore, in vitro TNF-alpha neutralization experiments and transfection of Hep G2 cells with an IL-8 construct confirmed that TNF-alpha and H2O2 directly stimulate IL-8 secretion. RT-PCR analyses indicated that chemokine secretion induced by these agents operates via increased gene expression. Furthermore, a variety of cytokine genes were found to be expressed by hepatocytes, including MCP-1, cytokine-induced neutrophil chemoattractant (CINC), and IL-6. Taken together, these studies indicate that hepatocytes respond to biologically relevant levels of common activators, including H2O2, to produce cytokines and chemokines that contribute to pathophysiologic and repair processes in the liver. PMID- 9726446 TI - Correlation of the appearance of anti-interferon antibodies during treatment and dimunition of efficacy: summary of an International Workshop on Anti-Interferon Antibodies. PMID- 9726444 TI - A study of filgrastim (rG-CSF) priming of etoposide/cisplatin in advanced non small cell lung cancer. AB - A previous phase II study (CALGB 9132) of etoposide/cisplatin + rG-CSF in patients with advanced non-small cell lung cancer (NSCLC) showed a marked difference in the absolute neutrophil count (ANC) nadirs between courses 1 and 2. Median ANC nadirs for courses 1 and 2 were 200 and 2500, respectively, suggesting a priming effect for rG-CSF. The present study was designed to determine whether rG-CSF given prior to the first cycle of chemotherapy would decrease the severity and duration of neutropenia. Twelve patients with stage IIIB or IV NSCLC and performance status 0-1 received rG-CSF 5 microg/kg for 5 consecutive days starting 7 days before treatment with etoposide 200 mg/m2 on days 1-3 and cisplatin 35 mg/m2 on days 1-3, repeated every 3 weeks. Patients also received rG CSF 5 microg/kg s.c. day 4 to postnadir ANC > 10,000. The median WBC nadir, ANC nadir, and platelet nadir after the first cycle of chemotherapy in the historical group (CALGB 9132) were 1300 cells/microl, 200 cells/microl, and 80,000 cells/microl, respectively. In the present study, the median WBC nadir, ANC nadir, and platelet nadir were 1300 cells/microl, 144 cells/microl, and 56,000 cells/microl, respectively. The median time for ANC to reach 10,000 cells/microl was 15 days in both the historical and the present study. For course 2, the WBC, ANC, platelet nadirs, and duration of grade 4 neutropenia were 2600, 1450, 70,000, and 0 days, respectively. This study failed to show a priming effect for rG-CSF when given in this dose and schedule. PMID- 9726447 TI - MR sialography? PMID- 9726448 TI - Understanding the natural history of saccular aneurysms. PMID- 9726449 TI - Transdural spinal cord herniation: acquired or developmental? PMID- 9726450 TI - When seeing double is not always bad. PMID- 9726451 TI - Clinical utility of positron emission tomography with 18F-fluorodeoxyglucose in detecting residual/recurrent squamous cell carcinoma of the head and neck. AB - PURPOSE: The use of positron emission tomography with 18F-fluorodeoxyglucose (FDG PET) to detect residual/recurrent squamous cell carcinoma of the head and neck has been tested only in small groups of patients. Our purpose, therefore, was to evaluate the ability of this technique to detect the presence of tumor at both primary and nodal sites in a large cohort of patients. METHODS: All patients referred for PET scanning over a 2.5-year period with a question of residual or recurrent squamous cell carcinoma of the head and neck were identified. Thirty five of 44 patients had sufficient follow-up to be meaningful to our analysis (range, 6-33 months). PET scans were interpreted visually with knowledge of the clinical history and correlative anatomic imaging findings. Detection of disease involving primary and nodal sites was assessed independently. Additionally, because each patient had been referred in an attempt to resolve a specific clinical problem, the usefulness of PET in accurately addressing these questions was assessed. RESULTS: At the primary site, sensitivity and specificity for residual/recurrent disease were 100% and 64%, respectively; for nodal disease, sensitivity and specificity were 93% and 77%, respectively. In helping to resolve the clinical question being asked, the positive predictive value of the test result was 65% and the negative predictive value was 91%. CONCLUSION: The high sensitivity and negative predictive value of PET scanning in our cohort of patients suggest an important role for this technique in the care of patients with suspected residual/recurrent head and neck carcinoma. The lower figures obtained for specificity and positive predictive value reflect the fact that increased FDG uptake may be due to either tumor or inflammation. PMID- 9726452 TI - PET and recurrent squamous cell carcinoma of the head and neck: a surgeon's view. PMID- 9726454 TI - Fluoroscopic MR of the pharynx in patients with obstructive sleep apnea. AB - PURPOSE: The purpose of our study was to introduce an ultrafast MR imaging technique of the pharynx as a diagnostic tool for viewing the mechanism of obstruction in patients with obstructive sleep apnea. METHODS: Six healthy volunteers and 16 patients with obstructive sleep apnea were examined on a 1.5-T whole-body imager using a circular polarized head coil. Ultrafast two-dimensional fast low-angle shot sequences were obtained in midsagittal and axial projections during transnasal shallow respiration at rest, during simulation of snoring, and during performance of the Muller maneuver. All patients underwent physical examination, transnasal fiberoptic endoscopy, and polysomnography. RESULTS: Five to six images were obtained per second with an in-plane resolution of 2.67 x 1.8 mm and 2.68 x 2.34 mm, allowing visualization of motion of the tongue, soft palate, uvula, and posterior pharyngeal surface. MR findings correlated well with results of clinical examination. The length of obstruction in the oropharynx, which cannot be ascertained by transnasal endoscopy of the pharynx, was clearly visible MR images. Differences between patients with obstructive sleep apnea and healthy subjects in terms of the degree of obstruction in the velopharynx and oropharynx depicted on MR images during the Muller maneuver were highly significant. CONCLUSION: We believe that ultrafast MR imaging is a reliable noninvasive method for use in the evaluation of obstructive sleep apnea. PMID- 9726453 TI - MR sialography in patients with Sjogren syndrome. AB - PURPOSE: The purpose of this study was to evaluate the effectiveness of MR sialography of the parotid gland ducts in the diagnosis and staging of Sjogren syndrome. METHODS: MR imaging was performed on a 1.5-T unit with a neck phased array coil. MR sialographic source images were obtained using a heavily T2 weighted fast spin-echo sequence with spectral fat suppression. All images were analyzed on the basis of maximum intensity projection reconstruction. Five healthy control subjects and 51 patients with definite Sjogren syndrome (43 with primary disease and eight with secondary disease) were examined with MR sialography. A labial gland biopsy was performed in all patients and histopathologic grading was done by means of focal scores. The findings of MR sialography were compared with the results of labial gland biopsy to determine the effectiveness of the technique in the diagnosis and staging of Sjogren syndrome. RESULTS: In all five control subjects, the main duct and the primary branching ducts of the parotid glands were clearly visible on MR sialographic images. In patients with Sjogren syndrome, a punctate, globular, cavitary, or destructive appearance was well seen within the parotid glands. Findings obtained at MR sialography correlated well with the results of labial gland biopsy. CONCLUSION: MR sialography has the potential to produce diagnostic findings in the parotid gland ducts of patients with Sjogren syndrome. Our results suggest that this method will augment and possibly replace X-ray sialography. PMID- 9726455 TI - Frequency and significance of a small distal ICA in carotid artery stenosis. AB - PURPOSE: Accurate calculation of the percentage of stenosis is crucial for identifying candidates for endarterectomy. Our goal was to quantify the reduction in diameter of the distal internal carotid artery (ICA) as a function of proximal ICA stenosis and to discuss the implications of distal ICA narrowing on the calculation of percentage of stenosis using the criteria of the North American Symptomatic Carotid Endarterectomy Trial (NASCET). METHODS: We retrospectively reviewed the carotid angiograms of 81 patients referred for evaluation of carotid stenosis. The caliber of the ICA stenosis and the diameters of the normal distal ICA, the common carotid artery, and the internal maxillary artery were remeasured with precision calipers. The percentage of stenosis derived from the NASCET criteria were compared with vessel diameter and with the difference in size of the ipsilateral and contralateral distal ICAs. We then recalculated the percentage of stenosis by substituting the presumed normal contralateral distal ICA diameter for the ipsilateral distal ICA diameter. RESULTS: In carotid arteries without significant stenosis (<70%), the distal ICA diameter measured 5.94+/-1.10 mm, but in vessels with severe stenosis (>70%), the distal ICA diameter measured 4.69+/-1.23 mm. After recalculation, four of 26 vessels were upgraded in classification from moderate (40% to 69%) to severe (>70%) stenosis. CONCLUSION: The diameter of the distal ICA begins to decrease when the proximal stenosis is 60% or greater. If the ICA distal to a stenosis is smaller than the contralateral ICA, recalculating the percentage of stenosis by substituting measurements of the contralateral distal ICA diameter may be warranted. PMID- 9726456 TI - CT of pilomatrixoma in the cheek. AB - Pilomatrixoma is an uncommon benign tumor arising from hair follicles. They occur most commonly in the head and neck region, and are usually found in girls during the first two decades of life. These tumors may contain calcification, which, when present, is helpful in suggesting the diagnosis. We present a classic case of pilomatrixoma in the cheek of a young woman. The tumor was documented on CT studies, which showed a subcutaneous, noninvasive mass with calcifications. PMID- 9726457 TI - Lateral retropharyngeal node metastasis from carcinoma of the upper gingiva and maxillary sinus. AB - Clinically unsuspected metastases to the lateral retropharyngeal nodes from carcinomas of the upper gingiva or maxillary sinus were found in five patients on follow-up CT examinations. Such uncommon metastases may follow the afferent lymphatic channels from the palate or pharyngeal region or arrive by retrograde lymphatics from positive neck nodes. Careful examination of lateral retropharyngeal nodes may be required in cancers of these primary sites. PMID- 9726458 TI - Carcinoma showing thymiclike differentiation (CASTLE tumor). AB - A 67-year-old woman had had a neck mass for 10 years, which recently increased in size. Sonographic, CT, and MR examinations showed a mass in the carotid and posterior spaces (triangle) extending from below the submandibular gland to the supraclavicular fossa, displacing the common carotid artery and the sternomastoid anteriorly. The mass was solid, noncalcified with lobulated outlines, hypoechoic on sonograms, of soft-tissue density on CT scans, isointense on T1-weighted MR images, hyperintense on T2-weighted MR images, and enhanced mildly after injection of contrast material on CT and MR studies. Histologic examination revealed a carcinoma showing thymiclike differentiation, a rare tumor of the neck and thyroid gland. PMID- 9726459 TI - Malleus bar as a rare cause of congenital malleus fixation: CT demonstration. AB - Among congenital ossicular anomalies without external ear atresia, malleus fixation is least common. We report a case of congenital malleus fixation by a bony bar connecting the malleus neck to the posterior tympanic wall, which was depicted on thin-section CT scans. PMID- 9726460 TI - Mastoid pneumocele causing atlantooccipital pneumatization. AB - A 49-year-old woman had a palpable mass in her occipital region. Plain radiographs and CT examination revealed extensive atlantooccipital pneumatization with findings consistent with the diagnosis of mastoid pneumocele. Decompression was achieved with placement of a myringotomy tube, resulting in prompt symptomatic relief. On a follow-up CT examination, the pneumatized areas had become opacified and new bone formation was present. PMID- 9726461 TI - MR-guided endoscopic sinus surgery. AB - We describe an interactive, intraoperative imaging-guided method for performing endoscopic sinus surgery (ESS) within a vertically open MR system. The procedure was performed with intraoperative imaging using a 0.5-T magnet with a 56-cm vertical gap. Interactive control of imaging planes was accomplished by optical tracking with two infrared light-emitting diodes mounted on an aspirator probe. The probe's position defined the location of the orthogonal imaging planes. Twelve patients with varying degrees of sinus disease underwent ESS with MR imaging guidance. Patients had acute and chronic sinusitis, nasal polyposis causing airway obstruction, or tumor requiring tissue biopsy. All procedures were performed with the patients under general anesthesia. The integration of endoscopy with optical tracking and intraoperative interactive imaging allowed localization of anatomic landmarks during ESS. No complications were encountered. PMID- 9726462 TI - Technique for arterial-phase contrast-enhanced three-dimensional MR angiography of the carotid and vertebral arteries. AB - Our goal was to evaluate whether contrast-enhanced three-dimensional MR angiography using the MR Smartprep technique would enable us to obtain arterial phase MR angiograms of the carotid and vertebral arteries. The study included 35 patients with suspected lesions of the neck in whom the MR Smartprep technique was used for MR angiography performed with a 1.5-T superconducting system. The tracker volume was placed primarily in the middle part of the right common carotid artery. The imaging volume was placed in a coronal direction to include the carotid and vertebral arteries from the aortic arch to the skull base. A centric phase-ordering scheme was used. Imaging times were 20 to 38 seconds for 14 patients and 11 to 16 seconds for 21 patients. By using a smaller tracker volume and an imaging time of less than 16 seconds, we were able to achieve a 100% successful triggering rate and to delineate selectively arterial-phase carotid and vertebral arteries with almost no venous contamination. Contract enhanced 3-D MR angiography with the MR Smartprep technique was useful for showing arterial-phase carotid and vertebral arteries selectively. PMID- 9726463 TI - Evaluation of Cushing disease: cavernous or inferior petrosal sinus sampling? PMID- 9726464 TI - Perspectives on multiple sclerosis. PMID- 9726465 TI - Dementia resulting from dural arteriovenous fistulas: the pathologic findings of venous hypertensive encephalopathy. AB - PURPOSE: Dural arteriovenous fistulas (DAVFs) are acquired arteriovenous shunts located within the dura. The highly variable natural history and symptomatology of DAVFs range from subjective bruit to intracranial hemorrhage and are related to the lesion's pattern of venous drainage and its effect on the drainage of adjacent brain. We examined the prevalence and features of DAVFs in patients with progressive dementia or encephalopathy. METHODS: The records and radiologic studies of 40 consecutive patients with DAVFs treated at our institution were reviewed. RESULTS: Five (12.5%) of 40 consecutive patients with DAVFs had encephalopathy or dementia. In each patient, high flow through the arteriovenous shunt combined with venous outflow obstruction caused impairment of cerebral venous drainage. Hemodynamically, the result was widespread venous hypertension causing diffuse ischemia and progressive dysfunction of brain parenchyma. Results of CT or MR imaging revealed abnormalities in each patient, reflecting the impaired parenchymal venous drainage. Pathologic findings in one patient confirmed the mechanism of cerebral dysfunction as venous hypertension. The hemodynamic mechanism and resulting abnormality appeared identical to that seen in progressive chronic myelopathy resulting from a spinal DAVF (Foix-Alajouanine syndrome). Remission of cognitive symptoms occurred in each patient after embolization. CONCLUSION: Venous hypertensive encephalopathy resulting from a DAVF should be considered a potentially reversible cause of vascular dementia in patients with progressive cognitive deficits. PMID- 9726466 TI - Venous hypertensive encephalopathy. PMID- 9726467 TI - CSF spaces in idiopathic normal pressure hydrocephalus: morphology and volumetry. AB - PURPOSE: Idiopathic normal pressure hydrocephalus (NPH) is an important cause of dementia in the elderly; however, idiopathic NPH is often difficult to differentiate from normal aging and vascular dementias in which brain atrophy with ventricular dilatation (hydrocephalus ex vacuo or central atrophy) is present. To elucidate the distinctive features of the distribution of CSF in idiopathic NPH, we used MR imaging to investigate the morphologic features and volume of the CSF space in patients with idiopathic NPH compared with those with other dementias. METHODS: We assessed the size of four CSF compartments (the ventricle, basal cistern, sylvian space, and suprasylvian subarachnoid space) in 11 shunt-responsive patients with idiopathic NPH by semiquantitative and volumetric analyses of coronal T1-weighted MR images. The results were compared with those in 11 age- and sex-matched patients with Alzheimer disease and in 11 patients with vascular dementia. RESULTS: In patients with idiopathic NPH, the CSF volume was significantly increased in the ventricles and decreased in the superior convexity and medial subarachnoid spaces as compared with patients with other dementias. The sylvian CSF volume in patients with idiopathic NPH was significantly greater than in patients with Alzheimer disease. The volume of the basal cistern was comparable among the three groups. In several patients with idiopathic NPH, focally dilated sulci were observed over the convexity or medial surface of the hemisphere. CONCLUSION: Our results indicate that findings of enlarged basal cisterns and sylvian fissures and of focally dilated sulci support, rather than exclude, the diagnosis of shunt-responsive idiopathic NPH and suggest that this condition is caused by a suprasylvian subarachnoid block. PMID- 9726468 TI - MR prediction of shunt response in NPH: CSF morphology versus physiology. PMID- 9726469 TI - Reversible hyperintensity of the anterior pituitary gland on T1-weighted MR images in a patient receiving temporary parenteral nutrition. AB - We report a patient with severe anorexia nervosa, treated with temporary total parenteral nutrition (TPN), in whom reversible hyperintensity of the anterior pituitary gland was seen on T1-weighted MR images. The anterior pituitary was isointense with white matter before TPN therapy and became markedly hyperintense after 3 months of treatment. The intensity normalized after TPN therapy was discontinued. The transient hyperintensity was also seen in the basal ganglia and dorsal brain stem. We believe the hyperintensity of the anterior pituitary may be attributed to the TPN therapy. PMID- 9726470 TI - Radiologic and pathologic findings of intracerebral schwannoma. AB - We report the radiologic and pathologic findings of an intracerebral schwannoma. MR imaging studies showed a superficially located cystic mass with an enhancing nodule and evidence of peritumoral edema or gliosis. PMID- 9726471 TI - Cat-scratch disease with an extraaxial mass. AB - CT and MR imaging of the brain and gallium-67 scintigraphy showed an enhancing, gallium-avid mass in the left middle cranial fossa of a 10-year-old girl. Craniotomy revealed an inflammatory mass related to the left trigeminal nerve. The lesion contained rodlike bacteria, and serologic tests were positive for cat scratch disease. Neurologic involvement in cat-scratch disease is uncommon, and the presence of organisms in neural tissue has not been reported. PMID- 9726472 TI - Real-time reconstruction and high-speed processing in functional MR imaging. AB - Access to fully processed activation maps in near real time during a functional MR examination enables run-to-run assessment of results. This is particularly useful in clinical studies, since the results of the functional MR examination can be ascertained before the patient leaves the MR suite, permitting interactive tailoring of the functional MR study. We describe how a real-time MR system can be customized to complete the following tasks in less than 3 minutes: obtain an 81-second acquisition of a multisection functional MR imaging time series using single-shot echo-planar imaging, perform image reconstruction, extract functional MR activation maps using cross-correlation and thresholding, and superimpose activation maps on previously acquired T1-weighted anatomic images. PMID- 9726473 TI - Evaluation of intraaneurysmal blood velocity by time-density curve analysis and digital subtraction angiography. AB - PURPOSE: Our purpose was to evaluate intraaneurysmal blood velocity by using time density curve analysis and digital subtraction angiography. METHODS: In 31 aneurysms, aneurysmal blood velocity was examined with digital subtraction angiography to determine mean transit time (MTF), peak density time (time to peak opacification) (PDT), and time to half-peak opacification (T1/2). Thirty frames per second were acquired, and the time-density curve was calculated. Regions of interest were drawn on the proximal parent artery, on the distal parent artery, and on the aneurysm itself. RESULTS: There was no significant difference in MTT of blood velocity in the proximal site on the parent artery, in the distal site on the parent artery, and in the aneurysm. Similarly, there was no significant difference in PDT in the parent artery, in the distal site on the parent artery, and in the aneurysm; nor was there a significant difference in T1/2 in the parent artery, in the distal site on the parent artery, and in the aneurysm; that is, intraaneurysmal blood velocity was similar to that in the parent artery. PDT and T1/2 of small aneurysms were faster than that of large aneurysms; that is, blood velocity of small aneurysms was faster than that of large aneurysms. CONCLUSION: Intraaneurysmal blood velocity in small aneurysms is similar to that in the parent artery; consequently, the central stream probably reaches the aneurysmal wall. Intraaneurysmal blood velocity in large aneurysms appears to be somewhat slower than that in small aneurysms. PMID- 9726474 TI - Histologic and morphologic comparison of experimental aneurysms with human intracranial aneurysms. AB - PURPOSE: Vein pouch aneurysms are the most commonly created experimental lesions in neuroendovascular research. We sought to determine whether an experimental aneurysm that is derived from a pancreatic elastase-digested arterial sac (EDASA) models the histology and morphology of human cerebral aneurysms more accurately than the vein pouch aneurysm does. METHODS: EDASAs were created in the common carotid arteries of four rabbits, and vein pouch aneurysms were created in the common carotid arteries of four pigs. Five recently ruptured human cerebral aneurysms were obtained at autopsy. Identical histologic preparations were made for all specimens, and a vascular pathologist performed blinded histologic analyses. Morphologic dimensions were measured with a micrometer at 40-fold magnification. RESULTS: In each human cerebral aneurysm, there was complete absence of internal elastic lamina and tunica media, and none showed evidence of mural inflammation or neointimal proliferation. Average wall thickness was 51 microm. All vein pouch aneurysms had a well-developed internal elastic lamina and tunica media, and all exhibited profound inflammation and neointimal proliferation. Average wall thickness was 290 microm. EDASAs were devoid of internal elastic lamina, their tunica medias were mildly atrophic, and the sac walls contained only mild inflammation and neointimal proliferation. Average wall thickness was 46 microm. CONCLUSIONS: EDASAs model the morphologic and histologic characteristics of human cerebral aneurysms more accurately than vein pouch aneurysms do. PMID- 9726475 TI - Detection of microemboli distal to cerebral aneurysms before and after therapeutic embolization. AB - PURPOSE: Recently developed interventional radiologic techniques, such as embolization with platinum coils, may induce thrombus formation within an aneurysm. The aim of the present study was to investigate the frequency of microemboli distal to untreated and treated cerebral aneurysms. METHODS: Among a total of 110 patients treated with platinum coil embolization, 35 patients (27 women and eight men, aged 50+/-10 years) who were at high risk of ischemic complications underwent emboli detection with a transcranial Doppler sonographic monitoring system. All patients were studied before and after coil embolization. The aneurysms were located at the internal carotid artery (n=14), the basilar artery (n=10), the middle cerebral artery (n=7), or the vertebral artery (n=4). Twenty-nine (85%) of 35 patients were monitored within 6 hours of the completion of treatment. RESULTS: Microemboli distal to the aneurysm were not detected in any of the patients before treatment. Microemboli were detected in 11 patients (31%) after embolization (mean, 16+/-21 per hour; range, 1-74 per hour). Microemboli were detected in five (71%) of seven patients in whom ischemic complications occurred after treatment, but in only six (21%) of 28 asymptomatic patients. This difference was statistically significant. The rate of occurrence of emboli in patients with ischemic complications (23+/-30 emboli per hour) was higher than in asymptomatic patients (10+/-7 emboli per hour), but this difference was not statistically significant. CONCLUSION: Microemboli were detected significantly more often in patients who suffered from cerebral ischemia after coil embolization of an intracranial aneurysm. This observation supports the definition of a high-risk group of patients with incomplete embolization or with a large-diameter, broad-neck aneurysm. The early detection of microemboli after treatment may be an indicator for excessive intraaneurysmal thrombus formation and could influence the decision for prophylactic treatment with heparin or aspirin. PMID- 9726476 TI - Intracranial pressure monitoring during intraarterial papaverine infusion for cerebral vasospasm. AB - PURPOSE: Intraarterial papaverine infusions are performed to reverse cerebral arterial vasospasm resulting from subarachnoid hemorrhage, but such infusions may lead to increases in intracranial pressure (ICP). This study was undertaken to determine when ICP monitoring is indicated during papaverine treatment. METHODS: Seventy-eight vessels were treated in 51 sessions in 28 patients with symptomatic vasospasm. ICP, papaverine doses, and infusion rates were recorded during treatment sessions. The procedural data, Hunt and Hess scores, Fisher grades, Glasgow Coma Scale scores, and ages for all subjects were reviewed and analyzed retrospectively. RESULTS: Baseline ICP ranged from 0 to 34 mm Hg. With typical papaverine doses of 300 mg per territory and infusion times ranging from 5 to 60 minutes per vessel, ICP increases above baseline during papaverine infusion ranged from 0 to 60 mm Hg. Significant (> or = 20 mm Hg) ICP increases during therapy were observed even in patients with low baseline ICP and with papaverine infused at the slowest rate. Patients with a baseline ICP of more than 15 mm Hg were much more likely to have significant ICP increases than were patients with a baseline ICP of 0 to 15 mm Hg. Hunt and Hess scores, Fisher grades, age, and Glasgow Coma Scale scores on admission and immediately before treatment did not correlate with ICP increases during papaverine infusion. Patients with ICP increases of more than 10 mm Hg during therapy were more likely to experience adverse clinical events than were patients with ICP increases of < or = 10 mm Hg. Reduction in the rate of papaverine infusion, or termination of infusion, resulted in reversal of drug-induced ICP elevation. CONCLUSION: ICP monitoring during intraarterial papaverine infusions for cerebral vasospasm is recommended for all patients and is particularly important for patients with elevated baseline ICP. Continuous ICP monitoring facilitates safe and time-efficient drug delivery. PMID- 9726477 TI - MR angiography with ultrashort echo time in cerebral aneurysms treated with Guglielmi detachable coils. AB - We evaluated a time-of-flight three-dimensional MR angiographic sequence with an ultrashort echo time for its ability to characterize the perfusional state of cerebral aneurysms that had been treated with Guglielmi detachable coils and to depict adjacent cerebral arteries. The results were compared with findings at conventional MR angiography and digital subtraction angiography. Adjacent vessels were seen better in 36% of patients imaged with the new technique. Both MR angiographic methods detected residual cerebral aneurysmal perfusion with a tendency to overestimate the patent portion of the aneurysm. PMID- 9726478 TI - Surgical transvenous embolization of a cortically draining carotid cavernous fistula via a vein of the sylvian fissure. AB - Percutaneous transvenous embolization is one of the most effective treatments of intracranial dural arteriovenous fistulas (AVFs) involving the dural sinuses. We present a unique case of surgical transvenous embolization in a 48-year-old man with a dural AVF of the cavernous sinus who presented with intracerebral hematoma. The dural AVF drained only into the vein of the sylvian fissure on angiography. Transvenous embolization via the vein of the sylvian fissure during craniotomy obliterated the AVF completely. PMID- 9726479 TI - Transdural spinal cord herniation: imaging and clinical spectra. AB - PURPOSE: Transdural herniation of the spinal cord is a rarely reported clinical entity, and many of the existing reports were published before the advent of MR imaging. We describe five current cases and compare them with findings in 25 cases reported in the literature to delineate the clinical and imaging spectra of transdural spinal cord herniation. METHODS: MR imaging, CT myelography, and conventional myelography were performed in five patients with transdural herniation of the spinal cord. These studies, along with clinical findings, are described. Intraoperative photographs are included for one case. The salient features of both the current and previously reported cases are summarized in tabular form. RESULTS: In three cases, transdural spinal cord herniation occurred posttraumatically, in one case the cause was iatrogenic and in the others the herniation occurred spontaneously. Imaging features not previously reported include dorsally directed herniations at thoracolumbar levels (two patients), apparent (lacking surgical confirmation) syringomeyelia (one case), a vertebral body nuclear trail sign (one case), and intramedullary hyperintensities on MR images (two cases). Clinical features not previously reported include unilateral pyramidal-sensory deficits (one case) and isolated unilateral pyramidal signs (one case). Clinical findings similar to previous reports include progressive paraparesis (two cases) and progressive Brown-Sequard syndrome (one case). CONCLUSION: Our five cases illustrate certain clinical and imaging findings not previously reported, and, together with the established features of the 25 cases in the literature, delineate the spectra of transdural spinal cord herniation. PMID- 9726480 TI - Spontaneous thoracic spinal cord herniation through an anterior dural defect. AB - A 44-year-old woman was examined for progressive left lower extremity weakness and spasticity. Thoracic spine MR imaging and CT myelography showed a ventral dural defect at T7-T8 with an extradural subarachnoid fluid collection and extradural herniation of the spinal cord. Intraoperative sonography confirmed the appropriate level for dural entry and the finding of spinal cord herniation. After reduction of the herniated spinal cord, the patient experienced gradual improvement in neurologic function. PMID- 9726481 TI - Brown-Sequard syndrome of the cervical spinal cord after chiropractic manipulation. AB - We report a case of increased signal in the left hemicord at the C4 level on T2 weighted MR images after chiropractic manipulation, consistent with contusion. The patient displayed clinical features of Brown-Sequard syndrome, which stabilized with immobilization and steroids. Follow-up imaging showed decreased cord swelling with persistent increased signal. After physical therapy, the patient regained strength on the left side, with residual decreased sensation to pain involving the right arm. PMID- 9726482 TI - Anterior spinal artery syndrome associated with severe stenosis of the vertebral artery. AB - We describe a 55-year-old man with quadriparesis and impaired pain and temperature sensation in whom T2-weighted MR images revealed a high-intensity lesion in the cord at C3-4. Angiography showed occlusion of the right vertebral artery and severe stenosis of the left vertebral artery. We concluded that the stenosis of the vertebral artery led to the anterior spinal artery syndrome and to a disturbance of consciousness. PMID- 9726483 TI - Are the brains of monozygotic twins similar? A three-dimensional MR study. AB - PURPOSE: The role of genetic mechanisms and the influence of environmental events in human brain development have been difficult to evaluate. The purpose of this study was to compare the cerebral cortical morphology and midline structures of monozygotic twin pairs using MR imaging. METHODS: Six observers, blinded to twin pairings, evaluated the 3-D renderings of the cortical surface and midline structures from MR images of seven monozygotic twin pairs. A morphometric analysis of the corpus callosum and of the distance between the anterior and posterior commissures was also performed. RESULTS: Despite surprising anatomic differences, the brains of the twin pairs were similar enough to enable the observers to distinguish twin pairs from unrelated subjects. Five of six observers correctly identified the brains of all seven twin pairs; the remaining observer failed to make a correct match in only one of seven pairs. Three of six observers identified the midline sagittal images of the related twins in all seven pairs, and the other three identified the related midline sagittal images in five of seven pairs. The results were statistically significant. CONCLUSION: Although the observed differences in morphologic characteristics between twins necessarily reflect nongenetic influences, the cortical patterns and midline structures of monozygotic twins probably are genetically similar. PMID- 9726484 TI - Imaging studies in a unique familial dysmyelinating disorder. AB - We report the imaging findings in five patients with a unique dysmyelinating disorder. MR studies of these infants showed obstructive hydrocephalus caused by mass effect produced by an enlarged cerebellum. The white matter of an enlarged cerebrum and cerebellum showed delayed myelination. Proton spectroscopy showed normal N-acetylaspartate (NAA) levels. While the dysmyelinating disorder was clearly differentiated from Canavan disease by an absence of elevated NAA and differing histopathologic findings and autosomal-dominant inheritance pattern, there were similarities to this disease in the presentation and, to some extent, in the initial imaging findings. PMID- 9726485 TI - MR imaging and localized proton MR spectroscopy in late infantile neuronal ceroid lipofuscinosis. AB - PURPOSE: Late juvenile neuronal ceroid lipofuscinosis (NCL) is a lysosomal neurodegenerative disorder caused by the accumulation of lipopigment in neurons. Our purpose was to characterize the MR imaging and spectroscopic findings in three children with late infantile NCL. METHODS: Three children with late infantile NCL and three age-matched control subjects were examined by MR imaging and by localized MR spectroscopy using echo times of 135 and 5. Normalized peak integral values were calculated for N-acetylaspartate (NAA), choline, creatine, myo-inositol, and glutamate/glutamine. RESULTS: MR imaging revealed volume loss of the CNS, most prominently in the cerebellum. The T2-weighted images showed a hypointense thalamus and hyperintense periventricular white matter. Proton MR spectra revealed progressive changes, with a reduction of NAA and an increase of myo-inositol and glutamate/glutamine. In long-standing late infantile NCL, myo inositol became the most prominent resonance. Lactate was not detectable. CONCLUSION: MR imaging in combination with proton MR spectroscopy can facilitate the diagnosis of late infantile NCL and help to differentiate NCL from other neurometabolic disorders, such as mitochondrial or peroxisomal encephalopathies. PMID- 9726486 TI - MR in a patient with Zellweger syndrome presenting without cortical or myelination abnormalities. PMID- 9726487 TI - Transformation of health care through innovative use of information technology: challenges for health and medical informatics education. AB - Information storage and processing continues to become increasingly important for health care, and offers enormous potential to be realised in the delivery of health care. Therefore, it is imperative that all health care professionals should learn skills and gain knowledge in the field of health informatics, or medical informatics, respectively. Working Group 1, Health and Medical Informatics Education, of the International Medical Informatics Association (IMIA WG1) seeks to advance the knowledge of how these skills are taught in courses for the various health care professions around the world, and includes physicians, nurses, administrators, and specialists in medical informatics. IMIA WG1 held its 6th International Conference on Health and Medical Education in Newcastle, Australia, in August 1997. The theme of the conference was 'Transformation of Healthcare through Innovative Use of Information Technology'. This special issue of the International Journal of Medical Informatics on Health and Medical Informatics Education contains selected papers presented at the conference. In addition to the central topic, Educating Health Care Professionals in Medical Informatics the topics telematics, distance education and computer based training were also discussed at the conference. PMID- 9726488 TI - Health and medical informatics education: perspectives for the next decade. AB - It is argued that the progress of information processing and information technology changes our societies. Examples are given that there is a significant economic relevance of information technology for medicine and healthcare and for the quality of healthcare as well. In order to adequately pursue the goal of 'Transforming healthcare through innovative use of information technology for the 21st century' (the topic of the 6th International Conference on Health and Medical Informatics Education and of this special issue of the International Journal of Medical Informatics), health professionals are needed who are well educated in health informatics or medical informatics, respectively. Raising the scope and the quality of education in the field of health and medical informatics would help to raise the quality and efficiency of healthcare. In this context the International Medical Informatics Association (IMIA) and its working group 1 (WG1) on Health and Medical Informatics Education can make a contribution by disseminating information and by elaborating recommendations on courses and programs in health and medical informatics. For this purpose IMIA WG1 has established a WWW site (http://www.imia.org/wg1) with information on health and medical informatics programs and courses. All teachers and institutions are encouraged to submit information about courses and programs offered and to set pointers to their own WWW sites. In addition, a mailing list was installed to facilitate communication between all persons involved in health and medical informatics education. For subscription, a message has to be sent to 'listserv@relay.urz.uni-heidelberg.de'. The body of the message should read 'SUBSCRIBE IMIA-WG1'. PMID- 9726489 TI - Health and medical informatics education for nurses and health service managers. AB - Health and Medical informatics is a discipline encompassing and combining aspects of all health, medical and informatics disciplines. Consequently, the topics to be covered in any educational program can vary considerably both in depth and breadth. Given that such programs need to meet the needs of a very diverse health professional workforce, educators need to develop curricula to suit specific target groups although common topic areas need to be included. This paper presents the state of play regarding nursing informatics education. It discusses informatics education for health service managers primarily in Australia through the use of a case study and compares these with some other similar programs. It then explores some of the issues encountered which are seen as impediments to the progression of health and medical informatics education, the most significant of which is traditional University organisational structures which do not readily facilitate multidisciplinary educational programs. PMID- 9726490 TI - Twenty five years of medical informatics education at Heidelberg/Heilbronn: discussion of a specialized curriculum for medical informatics. AB - The specialized university curriculum for medical informatics (MI) at the University of Heidelberg/School of Technology Heilbronn is one of the oldest educational approaches in the field of MI and has been successful now for 25 years with about 1000 graduates (Diplom-Informatikerin der Medizin or Diplom Informatiker der Medizin). It belongs to the category of dedicated master's programs for MI and is based on the concept of MI as a medical discipline of its own. It is oriented towards the total spectrum of MI ranging from health care economics, biosignal and medical image processing, medical documentation, to information and knowledge processing in medicine. It is a 4.5 years program with a strong emphasis on the methodological foundations of MI and on practical education in a number of specific laboratories. A total of 35 students are admitted each semester and in total about 440 students are enrolled. The faculty consists of 17 full-time members and about 25 part-time lecturers. The authors report on characteristics, structure and contents of the new fifth version of the curriculum and discuss the features of a specialized curriculum for MI with respect to the challenges for MI in the 21st century. PMID- 9726491 TI - HMI education for HIMs. AB - This paper analyses the curricula of the four Australian university programs for health information managers (HIMs) in relation to their coverage of health and medical informatics (HMI). The overlap between HIMs and HMIs should be increased through exchange of information at conferences such as this as well as communication and co-operation between the Schools of HIM and those offering health informatics related training at other Australian universities. PMID- 9726492 TI - Education and training of medical informatics in the medical curriculum. AB - The teaching of medical informatics is of importance for students in medicine and health care, realizing that they will be the health professionals of the future. Training in medical informatics is also of value for practicing clinicians who are overwhelmed by the avalanche of systems that are available on the market. Some examples of operational systems are presented here to indicate that health care has changed dramatically over the last decades. This paper intends to contribute to the drafting of IMIA guidelines for teaching medical informatics by (1) reporting on the experience at the Faculty of Medicine and Health Sciences of the Erasmus University Rotterdam as part of the curriculum, (2) reporting on the implementation of guidelines for teaching medical informatics in The Netherlands since these guidelines were drafted in 1986, and (3) by introducing the teaching material contained in the new Handbook of Medical Informatics and on its Web site. PMID- 9726493 TI - Integrated medical informatics with small group teaching in medical education. AB - National Taiwan University College of Medicine (NTUCM) introduced small groups of teaching and basic-clinical integrated courses for medical students in 1992. By using computer network and multimedia techniques, this study tried to overcome barriers to learning in small group teaching. The Department of Medical Informatics of NTUCM established campus networking and computer classrooms and provided Internet and intranet network services including mail, netnews, bulletin board systems (BBS), world wide web (WWW), gopher, ftp and local file servers. To implement an interactive learning environment, the authors first tried mail lists, newsgroups and BBS. Next an integrated learning system prototype on the WWW was developed to provide functions including online syllabus, discussion boards simulated to BBS, online talk, interactive case studies, virtual classroom with video on demand (VOD) and Internet medical resources. The results showed that after the medical students completed the required course of medical informatics and had good network access using a network to communicate with each other became a daily practice. In the future, the system will extend to the tutoring of clinical practice and continuing medical education. The authors expect a national medical education network and more international cooperation and exchange. PMID- 9726494 TI - Medical information processing: an interactive course for the Internet. AB - Medical students in Germany have to study the basics of medical information processing within a special curriculum which is part of the ecological course. This curriculum offers an introduction to principles of medical informatics. Starting with a conventional textbook, a computer-based training (CBT) program has been developed using the technologies of the internet and the World Wide Web (WWW). Features of the program include a well structured presentation of the information within the software and a high degree of interactivity. Early experiences suggest that this program enhances the learning in the domain of medical information processing. The program may be viewed via the URL: htfp.//www.med-rz.uni-sb.de/med_fak/imbei/pro jekt. PMID- 9726495 TI - Education in informatics in medicine and the health sciences--the need for relevance. AB - At the University of Newcastle, the 5-year undergraduate programme in Medicine has been developed as a problem-based, self-directed, fully integrated curriculum. Curricular integration involves not only the basic and clinical sciences, but also population medicine, critical reasoning and the development of a broad range of professional skills. Medical informatics has been seen as an increasingly important professional skill and the integrated nature of the curriculum has provided an appropriate setting for the introduction and continuing development of this component of the curriculum. Over the last 3 years this component has been developed to be incorporated into the curriculum for the health sciences within the faculty, becoming health informatics. Informatics for undergraduate students in medicine and allied health professions must be developed as a relevant and useful component of the curriculum. PMID- 9726496 TI - Medical informatics education by medical professors within their discipline. AB - A system for medical informatics education for medical students has been developed in the medical school. This paper describes the concept underlying the development of this system and its progressive outcomes over 8 years. In order to stimulate students to acquire computer-related knowledge and skills, this subject has been integrated into the course works of various medical subjects such as physiology. In addition, acquired knowledge and skills are evaluated within each subject by the production of reports for example, using computers. This provides a concrete example for students of the relevance of the information sciences to the solving of medical problems. A well equipped computer facility for the study of medicine also plays a significant role in inspiring student incentive. A computer room equipped with Macintosh computers was opened adjacent to the main medical library and is used in the same manner as the library, with books replaced by computers. In addition, all new students acquire their own Macintosh PowerBook. These various initiations have facilitated concept that the computer may be applied to medical problem solving at any time or place and may become as commonplace as a pen in daily medical practice. PMID- 9726497 TI - Integrating learning contexts in a medical informatics program--preparing for the introduction of PACS. AB - This paper analyzes the possibilities to extend learning contexts within a one year medical informatics educational program (MIP) at a Radiology Department in Sweden. The MIP was carried out within a theoretical framework based upon the integration of four learning contexts, which were inspired by Nonaka's theory of organizational knowledge creation. A summary of organizational knowledge creation theory and the ideas behind the learning contexts are presented. The main objective of the study was to investigate what would be the major benefits from the use of various learning contexts in a one-year medical informatics program? The MIP was found to form a basis for better learning conditions by increasing the flexibility, accommodating greater numbers of students as well as offering better possibilities for continuous learning. Evaluation of the MIP revealed that 98% of radiology department staff as compared to 39% of the intensive care unit staff, who had followed the hospital routine program, felt competent enough to independently use the functions of a new medical system. Although there are good reason to believe that the superior confidence for information technology (IT) is due to the integration of learning contexts, it can not be excluded that it may be due to other reasons also. PMID- 9726498 TI - Education of medical informatics in Bosnia and Herzegowina. AB - Time of information in which the authors live resulted in the increase of the amount of the information exponential growth of the new kind of knowledge, flourishing of the familiar ones and the appearance of the new sciences. Medical (health) informatics occupies the central place in all the segments of modern medicine in the past 30 years--in practical work, education and scientific research. In all that, computers have taken over the most important role and are used intensively for the development of the health information systems. Following activities develop within the area of health informatics: health-documentation, health-statistics, health-informatics and bio-medical, scientific and professional information. The pioneer in the development of the health statistics and informatics in Bosnia and Herzegovina (BiH) was Dr Evgenije Sherstnew, who was the Chief of Health Statistics in the Ministry of Health of BiH from 1946 1952, and who founded and led, from 1952 to the end of his life, the Department of Medical Documentation and Health Statistics of the Central Health Institute of BiH, the core around which a group of experts for the development of this field have gathered. In the eighties computers were intensively used as a tool for the processing medical data and with them the development of health information systems at the level of the outpatient-clinics, hospitals, clinical centers, as well as the integral information system of health, health insurance and the social security system of BiH began. Finally, Society for Medical Informatics of BiH, which as a professional association gathers experts in the area of health informatics, actively propagates this profession in the Republic, was founded. With reform of the lectures and curriculum at the medical faculty in Sarajevo, the course in 'Medical Informatics' has been in 1992. into the second semester, since it was assumed that an early insight into the principles of information along with studies of so called basic pre-clinic sciences, especially basics of information, would make things easier for the students the more informative education is in the course of their medical studies. The medical faculty in Sarajevo also established and accepted a course of health informatics and economics of post-graduate studies in 1979, of which the main objective is education of experts for work informatics jobs in health care system and services, especially for needs of the future information systems in BiH. PMID- 9726499 TI - Nursing students' blood pressure measurement following CD-ROM and conventional classroom instruction: a pilot study. AB - This paper describes a pilot investigation of first year nursing students' adherence to the recommended auscultatory blood pressure measurement procedure following three different forms of instruction; conventional classroom demonstration of the technique, a self instructional CD-ROM tutorial program, and a combination of both methods. This investigation was carried out over a 5-day time period using 27 students during normal teaching time. Students' adherence to the procedure was determined by observation using a performance checklist. Results suggest that the CD-ROM is no substitute for real life, hands on experience, although when used as an adjunct to traditional teaching methods, it can enhance learning. PMID- 9726500 TI - Use with care: possibilities and constraints offered by computers in nursing clinical education. AB - Funding constraints, demands for improved efficiency, low staff to student ratios and improved computer software (D. Wright, The use of multimedia computers and software in nurse education, Health Inf. 1 (3) (1995) 101-107.) all combine to produce a situation where a dramatic increase in the use of CBE for tertiary nurse education appears inevitable. In particular there is a very strong movement towards the development of nurse education materials able to be delivered via the World Wide Web (WWW). Given the speed and the extent of these developments it is essential that CBE developments are subject to adequate review and critique. In particular it is important that the pedagogical concepts under pinning the CBE are congruent with the technological innovation being implemented. This paper provides such a review and critique through the discussion of a project concerned with the integration of a computerised hospital nursing care planning database into the undergraduate nursing curriculum. The evaluation of the project not only raises many issues and concerns relevant to CBE in general, but also provides examples of the ways in which CBE can be used to link theory to practice. It also highlights the need for congruency between pedagogy, technology and student learning. Implications arising from the project are discussed at length, including the possibility for clinicians and academics to work collaboratively on database development. PMID- 9726501 TI - An international health and nursing informatics module for distance education. AB - This paper describes why a module about health and nursing informatics is a necessary component for nursing education. Several developments in society and health care force health providers to manage the large amount of health data adequately. A module about health and nursing informatics was developed in international cooperation by three schools of nursing from Germany, The Netherlands and the UK. The content and learning activities of the realized example module for distance learning are described. Future plans include making this course material available in different languages on the World Wide Web. PMID- 9726502 TI - Developing curriculum in nursing informatics in Europe. AB - The NIGHTINGALE Project (NIGHTINGALE Project: HC1109 DGXIII Contract and Technical Annex, European Commission, December 1995) which started on the 1st of January, 1996, after the approval of the European Commission, has a 36 month duration. It is essential in planning and implementing a strategy in training the nursing profession in using and applying healthcare information systems. NIGHTINGALE contributes towards the appropriate use of the developed telematics infrastructure across Europe by educating and training nurses in a harmonious way across Europe in the upcoming field of nursing informatics. NIGHTINGALE develops courseware material based on the curriculum development process using multimedia technologies. Computer based training software packages in nursing informatics will be the basis of the training material and the corresponding courses. CD-ROM based training and reference material will also be provided in the courses whereas the traditional booklets, teaching material and textbooks can also play an adequate role in training. NIGHTINGALE will disseminate all information and courseware material freely to all interested parties through the publications of the proceedings of the conferences, through the establishment of the world wide web (WWW) server in nursing informatics for Europe (http://www.dn.uoa.gr/nightingale), which will become a depository of nursing information knowledge across Europe as well as a dissemination node of nursing informatics throughout the European members states for the benefit and welfare of the European citizen. PMID- 9726503 TI - Computers in the Faculty of Health Science--5 years on. AB - In 1989, the Faculty of Health Science at Central Queensland University (CQU) committed itself to a major instructional development project for its nursing education program. Partially supported by a National Priority (Reserve) Fund (NPRF) grant, the Faculty undertook a multi-year project to develop computer based learning materials. The success of that project was mixed. However, by 1993 staff and students within the Faculty were using computers regularly, many of the staff were involved in developing computer-based instructional materials, and some staff were using available computer-based tools to extend the scope of the standard Health Science materials developed under the project. As well, the Faculty was committed to using Faculty funds for technical support staff to maintain its computer infrastructure and to assist both staff and students in using that infrastructure. The authors review the original NPRF project, then explore the Faculty's continued development and use of computer-based learning materials through an examination of student and staff evaluations, personal recollections, and subsequent projects. This paper demonstrates that the NPRF project spawned a number of other computer-based developments, including the offering of health informatics degree programs at the undergraduate and postgraduate levels, the development of the Faculty's current Internet site and a CD-ROM based interactive multimedia project for diabetes education. It also demonstrates that moderate levels of funding can maintain a computer-literate Faculty which views computers as essential tools for learning. PMID- 9726504 TI - The teaching of computer programming and digital image processing in radiography. AB - The increased use of digital processing techniques in Medical Radiations imaging modalities, along with the rapid advance in information technology has resulted in a significant change in the delivery of radiographic teaching programs. This paper details a methodology used to concurrently educate radiographers in both computer programming and image processing. The students learn to program in visual basic applications (VBA), and the programming skills are contextualised by requiring the students to write a digital subtraction angiography (DSA) package. Program code generation and image presentation interface is undertaken by the spreadsheet Microsoft Excel. The user-friendly nature of this common interface enables all students to readily begin program creation. The teaching of programming and image processing skills by this method may be readily generalised to other vocational fields where digital image manipulation is a professional requirement. PMID- 9726505 TI - Medical informatics training programme to support the Romanian health care management information system. AB - The Health Management Information System (HMIS) project initiated by the Romanian Ministry of Health as a component of the healthcare reform is aiming to ensure the technical, functional and operative support for: (i) a better overview of the population health status, of the medical care needs and of the Health System performances; (ii) the improvement of the resource allocation and consumption; and (iii) reform support. This system is supposed to assure a better information flow from the lower to the upper levels of the healthcare network by help of a modern IT support. In the first development stage the system is planned to link the Ministry of Health with the 41 District Health Authorities (DHAs) and with more than 200 pilot health units. The implementation of such a large system raises serious problems of acceptability and a thorough training programme for both technical staff and end users must be considered in order to face this challenge. PMID- 9726506 TI - The graduate training in medical information sciences in the Academic Medical Centre at the University of Amsterdam. AB - Since its inception in 1987, the 4-year Medical Information Sciences (MIS) curriculum at the Academic Medical Centre (AMC), Amsterdam has gone through several major changes. The present curriculum started in 1994. The course takes 4 years, the first 3 years are programmed in integrated modules of 7 weeks in duration each. In these modules much attention is given to interactive teaching, problem based learning and private study. Typical for the Amsterdam curriculum is a strong emphasis on the role and significance of data and information in health care and its management. The authors see information technology per se as auxiliary to this orientation. Presently, about 150 students follow the courses. PMID- 9726507 TI - Assessing the progress of the M.Sc. course in health informatics under the ERASMUS programme. AB - Health informatics is an emerging and important multi-disciplinary field that involves, informatics but also medicine, nursing, engineering, biology and other related subjects. A co-ordination of this field at a postgraduate level becomes important now in Europe where other European Community programs such as the 'Telematics for Health Care' will require at the Fourth Framework Programme (94 99) adequate human resources of higher potential and knowledge. This European M.Sc. course meets all the above objectives. The Curriculum was developed according to the results of the ERASMUS Workshop, which was held in Athens on 13 15 September 1990 under the ERASMUS Contract number ICP-90-G-0009/12. The implementation now runs under the contract ICP-95-G-1038/12. The 6-year evaluation of the course based both on staff and student evaluation proved that the M.Sc. course in health informatics has been successful. PMID- 9726508 TI - The need for a coherent curriculum and supported infrastructure in health informatics education--the HEALNet experience. Health Evidence Application and Linkage Network. AB - The Canadian National Centres of Excellence Network, Health Evidence Application and Linkage Network (HEALNet) is a network linking universities, the private sector and other public sector organisations in supporting research and fostering commercial opportunities. Its aim is to develop high quality health care decision support methodologies and tools that support the transfer of research evidence into practice. The HEALNet education program aims to support the training of highly qualified personnel as well as the development of educational approaches so that health personnel can better exploit such methodologies and decision support tools given the new opportunities of information technology. There is a need to draw from the different types of expertise within HEALNet to provide an integrated curriculum. At the same time, there is a concern regarding the lack of a coherent infrastructure in health informatics in many Canadian medical schools. PMID- 9726510 TI - Education and training in health informatics: the IT-EDUCTRA project. AB - In this contribution both the EDUCTRA project of the European Advanced Informatics in Medicine Programme and the IT-EDUCTRA project of the Telematics Applications Programme (Health Sector) are described. EDUCTRA had as aim to investigate which gaps in knowledge health professionals have with respect to health informatics and to suggest ways to remedy this. It was assumed that health professionals had a basic understanding of health informatics and that additional educational material only had to cover the knowledge necessary for appreciating the new products coming from the AIM programme. A state-of-the-art survey revealed that the knowledge of health professionals with respect to health informatics was deplorable. Guidelines for curricula were therefore proposed to enable potential teachers to design courses. IT-EDUCTRA is a continuation of the EDUCTRA project. It has as aim to create learning materials covering a broad area of health informatics. PMID- 9726509 TI - Different approaches to the tasks of educating and training information systems professionals, within the National Health Service (UK). AB - In 1994, La Sainte Union College of Higher Education (LSU) developed an MSc in Health Informatics course, in conjunction with Southampton University NHS Trust (SUHT). The original part-time, 1 day per week mode of delivery has since been broadened to include a distance leaning route and recently a block release mode, by which students combine usage of the distance learning materials with attendance in College for an intensive 2-day taught element. Because the course was designed in close co-operation with a major teaching hospital, it has always been 'market led' to meet the needs both of the individual students and of the organisations that they work for. At the same time, students acquire a quality assured qualification from a premier UK university, a qualification that holds credence outside the National Health Service (NHS). At the same time as LSU and SUHT were developing the MSc in Health Informatics, the UK NHS Training Division (NHSTD) started to promote a professional qualification for health service professionals. the so-called 'Statement of Recognition' (SoR). In contrast to the academic format of an MSc, the SoR was not a formal course, but a combination of modules designed to help candidates demonstrate their competence and achievement at work by portfolio evidence. This approach has national standing throughout the UK in a set of qualifications known as NVQs (National Vocational Qualifications). The NHSTD, through its successor, the Institute of Health Care Development (IHCD), has further refined this competency based model, culminating in the launch in 1996 of the Diploma and Advanced Diploma in Information and Technology (Health). Professionals within the area of Information Management and Technology (IM&T) in the NHS now have the alternatives of an academic or a competency route to achieve their goals. This paper traces the development of and the relationship between, these two approaches to the educational and training of healthcare professionals. It will illustrate the shift from Information Technology (IT) to Information Management skills, which is a pre-requisite to satisfy the changing needs of information users. It will also consider how a single Master's course can lead to a range of courses which meet the needs of professionals at various levels. Finally, it makes some recommendations for future developments of the programmes, suggestions which may have some relevance outside of the UK. PMID- 9726511 TI - Determinants of microcomputer technology use: implications for education and training of health staff. AB - In hospitals and other Healthcare settings, increasingly, hands-on computer use is becoming an important behaviour for effective job performance. The literature has identified differences that relate to computer use between occupational categories in health services. The objectives of this study were to identify factors that determine computer acceptance among occupational groups in Community Health and to predict the factors that relate to computer use. A survey was administered to all Community Health staff in one health service area. Health administrators were found to have a significantly higher training in computers, a higher frequency of use and a higher level of skill for both applications (word processing (WP) and database (DB)) than nurses. The results of a regression analysis shows that about 55% of the variation in the use of WP is explained by computer skills, perceived usefulness (PU) and designation. In the case of DB use, PU was the only significant predictor explaining 53% of the variation. Both level of education and prior training were not significant predictors. The implication for health informatics education (and service training) of these findings is that, in the workplace, health professionals would use computers when they perceive it to be useful for performance in their jobs. PMID- 9726513 TI - Strategic planning of the master programme in health informatics at Aalborg University: targeting and updating the programme, to meet explicit customer needs. AB - Education is essentially giving people new skills and qualifications to fulfil certain tasks. In planning and managing educational programmes it is crucial to know what skills and what qualifications are needed to carry out the tasks in question, not to mention the importance of knowing what tasks are relevant to carry out. The programme in health informatics at Aalborg University produces health informatics professionals. The students are developing skills in solving informatics problems in health care organisations. The programme has been running for 3 years now and to maintain the perception of the aim for the programme a number of activities have been launched. In the following, the programme will be presented, the activities to obtain information on how to keep the programme targeted and updated will be described and the changes that are going to be introduced will be outlined. PMID- 9726512 TI - Teaching the fundamentals of information systems management in health care. Lecture and practical training for students of medical informatics. AB - For the management of information systems in health care, it is important that projects are systematically planned and carried out. This is a major task for medical informatics professionals which should be taught in a medical informatics curriculum. In the respective lecture in the Heidelberg/Heilbronn medical informatics curriculum, we teach fundamentals of the management of information systems and of projects. The examples of the lecture are taken from hospital information systems. Furthermore, we have developed a 5-step method for the systematic, goal-oriented planning of projects. The lecture is complemented by a comprehensive practical training, so that the methods taught can be applied to a particular, relevant problem of the Heidelberg University Hospital. PMID- 9726514 TI - The university training programs in Austria. AB - A description is given of Austria's currently operational framework of training opportunities and post-doctoral courses in Medical Informatics. The description includes a brief outline of the currently valid curriculum in informatics and the methods on how to specialise in medical informatics within this framework. PMID- 9726515 TI - On development of medical informatics education via European cooperation. AB - In this paper, we show different activities of the European Center for Medical Informatics, Statistics and Epidemiology (EuroMISE Center) of Charles University and Academy of Sciences in the field of medical informatics, statistics and epidemiology education and training. The development of these activities started within the TEMPUS-PHARE project in 1993 and they are continuing with the support of another project, particularly the IT EDUCTRA (Information Technologies Education and Training) (Fourth Framework Programme) project. New approaches using the Internet as well as newly developed programmes are described. PMID- 9726516 TI - Medical problem based learning supported by intranet technology: a natural student centred approach. AB - In response to the explosion in medical information, there have been considerable recent changes in medical curriculum development. The move to problem based learning (PBL) is, in part, a result of these changes. The Faculty of Medicine at the University of Sydney has exploited a WWW based intranet for the development, delivery, management and evaluation of it's problem based, graduate medical program (GMP). This system has been employed to develop the 72 medical problems that contribute to the first two years of the GMP. The activities of more than 400 members of the faculty have been coordinated using the intranet to develop the wide range of resources to support learning in the program. Daily management of the curriculum is also enabled using Web site posting of bulletins, e-mail and ongoing development of technology training. Coupled with the PBL problems is a formative assessment system that provides questions and feedback that cover the whole range of learning topics. Part of the student and staff evaluation is supported both informally and formally through the use of a 'Feedback' button on each web page and web delivered structured formal evaluations, respectively. PMID- 9726517 TI - Validation of computer-mediated open-ended standardized patient assessments. AB - Quantitative information regarding the development of students' clinical reasoning skills is valuable in assessing third- and fourth-year medical students' clinical knowledge. Standardized patient cases are often used to obtain this quantified information. These cases typically involve a post-patient examination consisting of a series of closed-ended multiple-choice questions. Many medical educators question whether the results from the closed-ended multiple-choice questions truly reflect students' clinical knowledge and reasoning proficiency. Since 1995, the Kirksville College of Osteopathic Medicine (KCOM) has used a computer program, SOAP Note Plus, for standardized patient assessment of students to automate the post-encounter documentation and evaluation. This paper describes the development of the SOAP Note Plus program and a validation study which was conducted to determine the empirical association between the computer-mediated closed-ended and open-ended post-encounter standardized patient assessments. Correlation and GENOVA statistics were used in the analysis of the performance of third-year medical students on the closed ended and the open-ended assessments following standardized patient encounters and the relationship to their undergraduate GPA, first 2-year medical school GPA and the actual clinical rotation evaluations. The initial results show the positive relationship between the open-ended assessment and the actual clinical rotation evaluations. undergraduate GPA and the first two-years of medical school GPA. PMID- 9726518 TI - Development and validation of an evaluation tool for multimedia resources in health education. AB - The last decade saw a rapid increase in the use of multimedia in health education. Easy availability, accessibility, low cost of technological resources and the expanding body of research on the role of multimedia in student learning, among others, have all contributed to this increase in usage. Since one of the roles of educators is to assess and select learning resources based on curriculum goals and student needs, the development of standardized methods for multimedia evaluation becomes vital. To the learner, it is important for reviews of the quality of the resource to be readily available. An evaluation too was developed based on the recognition of this need. The validity of the tool was tested using experts in technology and education. Reliability was determined using faculty and students who reviewed the same software, using the tool. In addition, graduate students reviewed two versions of a nursing program, of varying quality. The results indicate that the tool is reliable and valid. It is envisaged that this tool can be utilized by health educators for evaluating multimedia resources and setting up a much needed clearinghouse for health education resources. PMID- 9726519 TI - Production of CAL-programs in medicine, odontology and veterinary medicine in Sweden. AB - At the recommendation of the Swedish Government, the Council for the Renewal of Undergraduate Education was established in 1990. In 1993 the Council was declared a permanent National Agency by Swedish Parliament and became part of the newly established National Agency for Higher Education in 1995. The purpose of the Council for Renewal of Undergraduate Education is to promote and support endeavors to develop quality and renewal of undergraduate education. In particular the council awards grants to development activities. Once a year, teachers at Swedish universities, university colleges and professional schools can apply for funding. Applications are accepted for projects directed towards undergraduate education in all disciplines. The Council selects 15-20 projects and each project is funded for 1-3 years. An advisory group--MEDCAL (Computer Assisted Learning (CAL) in MEDicine, Odontology and Veterinary Medicine)- consisting of representatives from all universities supports the Council with registration and evaluation of programs, offers their opinions on the production of CAL and collaborates with similar organizations in other countries, e.g. Australia, Denmark, Germany, Great Britain and USA. In all, 12 projects within the frame of MEDCAL will be reported. PMID- 9726520 TI - Multimedia system based on programmed instruction in medical genetics: construction and evaluation. AB - A multimedia system used as an auxiliary didactic tool for teaching medical genetics, HGEN, is based on non-linear programmed instruction and multimedia. HGEN was implemented in layers for PC compatible using MULTIMEDIA TOOLBOOK and DELPHI. It includes basic medical genetics concepts (inheritance patterns and cytogenetics) and it is based on multiple choice questions with images, diagrams and animations. The student-program interaction occurs by choosing an alternative in a question and receiving a specific answer as feedback and additional information. Links send the students to a glossary, to short descriptions of diseases used as examples, or to references for further studies. In order to evaluate the performance of the HGEN the authors used two questionnaires: (a) about the students' background in using computers; and (b) the system efficiency as a didactic tool, the software quality and user satisfaction. HGEN was used by 63 students from three medical schools and it was considered efficient as a learning tool (100%). Furthermore the implementation of a navigation map for frequencies analysis of followed path enable the study to detect structure and content inadequacies that could be correct for version 1.1 and proved to be an efficient tool to optimize a didactic software. PMID- 9726521 TI - The implementation of an integrated on-line health education system at RMIT. AB - The Faculty of Biomedical and Health Sciences at RMIT has been developing an on line health education system using a systems thinking approach, to create a learning environment whose basis is supported by Information Technology (IT). The centre-piece of this system is the Faculty Learning Centre, which has been created, both in space and layout, to promote collaborative learning between the students, so that the educator is physically assimilated with the student body. This facility is supplemented by the Faculty WWW server, which has been the main vehicle for course material dissemination to students. To ensure an effective on line teaching environment, the position of an on-line facilitator has been created, whose responsibilities include both the continual evaluation of the system and the implementation of appropriate system changes. Aspects have included the production of a staff development training program and extensive user documentation. This paper discusses the systems thinking approach used to implement this integrated on-line system, and the establishment of explicit educational rationales in the use of IT to support learning strategies. Some examples of the on-line educational programs are also presented. PMID- 9726522 TI - Challenges for delivering healthcare education through telematics. AB - Within the context of the education of professional healthcare providers, the authors give an overview of the challenges faced by those wishing to introduce telematics as both a mechanism for content delivery and as subject content itself. After presenting a brief overview of the current state of telematics applications to healthcare education, focusing on the European sphere, the authors outline the challenges before discussing the collaborative and communicative issues in detail. The authors conclude by suggesting that, while the authors believe that telematics is a necessary direction for the future development of healthcare education for professionals, the collaboration and communication challenges are of greater importance than the technical and policy challenges and that there is a need to educate the majority of educators, based in the experiences of the enthusiasts. PMID- 9726523 TI - Distance learning, problem based learning and dynamic knowledge networks. AB - This paper is an attempt to develop a distance learning model grounded upon a strict integration of problem based learning (PBL), dynamic knowledge networks (DKN) and web tools, such as hypermedia documents, synchronous and asynchronous communication facilities, etc. The main objective is to develop a theory of distance learning based upon the idea that learning is a highly dynamic cognitive process aimed at connecting different concepts in a network of mutually supporting concepts. Moreover, this process is supposed to be the result of a social interaction that has to be facilitated by the web. The model was tested by creating a virtual classroom of medical and nursing students and activating a learning session on the concept of knowledge representation in health sciences. PMID- 9726524 TI - The current and future role of the Internet in patient education. AB - The Internet technology known as the World Wide Web is rapidly emerging as the most powerful medium of mass communication this century and it can be harnessed to dispense global, cost-effective, high-quality, multimedia patient education material. This paper reviews how the Internet has progressed from delivering simple static, text-based material to sophisticated interactive Web sites based on CGI Technology. Interactive Web sites can be used to deliver health assessment questionnaires and Web-based decision-support systems can give patients advice on the emergency management of acute medical problems. The advantages and drawbacks of this new technology, including information regulation and quality are discussed. The role of the Hospital Intranet as a patient education resource is described. The paper concludes by illustrating how patients can appreciate the 3 D structure of bones and organs using virtual reality in a VRML Web environment. PMID- 9726525 TI - Maximising the world wide web for high quality educational and clinical support to health and medical professionals in rural areas. AB - Attracting and retaining medical and allied health professionals to rural and remote regions is a perennial problem, world-wide. It is exacerbated by the associated problem of providing high quality, cost-effective educational and clinical support to health professionals and trainees located in such areas. The world wide web is being used as a tool to address these twin problems to provide a means of communication, interaction and educational and clinical support to geographically dispersed and isolated pools of users. An interactive electronic notice board has been developed as a clinical problems' discussion forum to act as a venue for critical debate about clinical issues and to provide a searchable archive of information to assist in the management of clinical problems. A description of the forum will demonstrate the potential of the world wide web as a learning and a clinical support tool. PMID- 9726526 TI - Small worlds and medical expertise: implications for medical cognition and knowledge engineering. AB - This paper proposes and defends the small worlds hypothesis, which states that expert physicians organize diagnostic knowledge on the basis of similarities between disease categories, forming 'small worlds' consisting of small subsets of diseases and their distinguishing features. Examining existing data from several previous studies, the authors provide support for the small worlds hypothesis and for a characterization of the process of expert medical diagnostic reasoning as a succession of limited comparisons involving related diagnostic hypotheses. In one study, subjects were presented clinical endocrine cases one statement at a time and were prompted to think aloud after presentation of each statement. A combination of discourse and protocol analysis techniques were used to investigate hypothesis generation and evaluation. In another study, dialogues from doctor-patient interviews were examined. It was found that expert subjects rapidly select relatively small sets of plausible diagnostic hypotheses (small worlds) and focus on the most relevant medical findings that distinguish among the diseases in such small worlds. Results from both studies indicate that expert physicians use efficient strategies for discriminating among these alternative hypotheses in a stepwise process. In contrast, non-experts often generate large numbers of possible diagnostic hypotheses, belonging to widely differing disease categories. The results provide empirical support for the theoretical basis of small worlds. The implications of these results for the study of medical expertise and knowledge engineering are discussed, as well as considerations for the development of decision support systems. PMID- 9726527 TI - A Dutch medical language processor: part II: evaluation. AB - This paper provides a preliminary evaluation of a general Dutch medical language processor (DMLP). Four examples of different potential applications (based on different linguistic modules) are presented, each with its own evaluation method. Finally, a critical review of the used evaluation methods is offered according to the state of the art in medical language processing. PMID- 9726529 TI - An overview of the effect of computer-assisted management of anticoagulant therapy on the quality of anticoagulation. AB - Risks and benefits of anticoagulant therapy depend directly of the quality of anticoagulation. We carried out a meta-analysis of published randomized trials to assess the overall effectiveness of computer-assisted prescription systems on the quality of anticoagulation. Randomized controlled trials were identified through electronic searches of the Medline database (1966-1997) and systematic analyses of the references of articles. Two investigators selected relevant papers and summarized data from the studies. The outcome variable was the proportion of days within the target range of anticoagulation. A pooled estimate of the common odds ratio of being in the target range and its confidence interval was obtained by the Mantel-Haenszel method. Nine trials having included 1336 patients were identified. Computer systems were based on a pharmacokinetic-pharmacodynamic model and a bayesian prediction method. Most of them concerned the oral anticoagulant warfarin. The global odds ratio of being in the target range was 1.29 [95% CI: 1.17-1.49], thus meaning that the use of a computer for anticoagulation optimization increased by 29% the proportion of visits where patients were appropriately treated. The proportion of clinical events was too low for allowing a summary analysis, but major hemorrhages tended to be less frequent among patients of the computer groups than among patients of the control groups (2.0 versus 3.9%). Evidence from randomized controlled trials supports the effectiveness of computer-aided anticoagulant prescription. Widespread use of these systems in ambulatory care could increase the benefit/risk ratio of anticoagulant treatment at a low cost. PMID- 9726528 TI - ARIANE: integration of information databases within a hospital intranet. AB - Large information systems handle massive volume of data stored in heterogeneous sources. Each server has its own model of representation of concepts with regard to its aims. One of the main problems end-users encounter when accessing different servers is to match their own viewpoint on biomedical concepts with the various representations that are made in the databases servers. The aim of the project ARIANE is to provide end-users with easy-to-use and natural means to access and query heterogeneous information databases. The objectives of this research work consist in building a conceptual interface by means of the Internet technology inside an enterprise Intranet and to propose a method to realize it. This method is based on the knowledge sources provided by the Unified Medical Language System (UMLS) project of the US National Library of Medicine. Experiments concern queries to three different information servers: PubMed, a Medline server of the NLM; Theriaque, a French database on drugs implemented in the Hospital Intranet; and a Web site dedicated to Internet resources in gastroenterology and nutrition, located at the Faculty of Medicine of Nice (France). Accessing to each of these servers is different according to the kind of information delivered and according to the technology used to query it. Dealing with health care professional workstation, the authors introduced in the ARIANE project quality criteria in order to attempt a homogeneous and efficient way to build a query system able to be integrated in existing information systems and to integrate existing and new information sources. PMID- 9726530 TI - Integration of a blood pressure controller and an infusion toolbox system using client-server technology. AB - This paper shortly describes an Infusion Toolbox and a blood pressure (BP) control application. It explains how we applied an agent model and client-server technology to integrate them. We show that by using this framework and object oriented technologies the necessary changes to the existing applications are reduced. Many elements of the tested original systems could be re-used, which enhances safe development. PMID- 9726531 TI - A general classification of U-shaped dose-response relationships in toxicology and their mechanistic foundations. AB - The development of a comprehensive database of chemical hormetic responses (i.e., U- or inverted U-shaped dose-response relationships) using objective a priori study design, statistical and study replication criteria has recently been reported. An assessment of this database reveals the existence of a wide range of hormetic dose-response relationships including those demonstrating a direct stimulation or an overcompensation response to a disruption of homeostasis. These two broad types of hormetic responses are affected by temporal factors and display unique patterns of dose-range stimulation, magnitude of stimulatory response and relationship of the maximum stimulatory response to the NOAEL. A general classification of U-shaped dose-response relationships is proposed to provide a more organized framework to evaluate the highly distinctive and diverse hormetic responses within the context of establishing underlying biological mechanisms and exploring risk assessment implications. PMID- 9726532 TI - Effects on developmental landmarks and reproductive capability of 3,3',4,4' tetrachlorobiphenyl and 3,3',4,4',5-pentachlorobiphenyl in offspring of rats exposed during pregnancy. AB - 1. Pregnant Wistar rats were treated orally with a single dose of 100 microg 3,3',4,4'-tetrachlorobiphenyl (PCB 77)/kg b.w. or 10 microg 3,3',4,4',5 pentachlorobiphenyl (PCB 126)/kg b.w. on day 15 of pregnancy. The control rats received peanut oil at the same day. Developmental landmarks were assessed in all offspring rats and reproductive effects of PCB 77 and PCB 126 on male offspring were studied on postnatal day 65 (at puberty) and on postnatal day 140 (at adulthood). 2. The ano-genital distance as well as the ratio ano-genital distance to body length was reduced in male pups of the PCB 126 group and the age at vaginal opening was significantly delayed in the female pups. 3. Testis, brain weights and daily sperm production were permanently increased and seminal vesicle weights were decreased in male offspring of the PCB 77 group. In male rats of PCB 126 group, the brain weights were permanently increased and ventral prostate weights permanently reduced. In both PCB groups, however, serum testosterone concentration was reduced only at adulthood. Additionally, the male rats of the PCB 126 group showed alterations in sexual behavior. In these rats the number of mounts with intromissions was significantly increased. 4. The results of this study show that PCB 126 elicits some TCDD-like reproductive effects after in utero exposure, while the reproductive effects of in utero exposure to PCB 77 on male offspring may be attributed to the neonatal hypothyroidism induced by the substance during early fetal development. Further studies using multiple doses and providing thyroid hormone data will be necessary to support this hypothesis. PMID- 9726533 TI - Protection of human upper respiratory tract cell lines against sulphur mustard toxicity by hexamethylenetetramine (HMT). AB - 1. Sulphur mustard ('mustard gas', HD) is a highly toxic chemical warfare agent which affects the skin and respiratory tract. The primary targets of inhaled HD are the epithelia of the upper respiratory tract. Hexamethylenetetramine (HMT) has been shown to protect human lung cells against HD toxicity and has also been shown to be effective in vivo against the chemical warfare agent phosgene. The ability of HMT to protect against the toxicity of HD was investigated in the human upper respiratory tract cell lines BEAS-2B and RPMI 2650. 2. HD was highly toxic to both cell lines, with LC50 values of 15-30 microM. HMT, at a concentration of 10 mM, was shown to protect the cell lines against the toxic effects of 20 microM and 40 microM HD. Results demonstrated that it was necessary for HMT to be in situ at the time of exposure to HD for effective cytoprotection. No protection was seen when cells were treated with HMT following exposure to HD, or where HMT was removed prior to HD exposure. 3. Results suggest that HMT may be effective prophylaxis for exposure to HD by inhalation. PMID- 9726535 TI - Protection of human upper respiratory tract cell lines against sulphur mustard toxicity by glutathione esters. AB - 1. Human and animal lung cells have been used successfully to model the toxic effects of inhaled sulphur mustard (HD). The epithelia of the upper respiratory tract are, however, the primary targets of inhaled HD. The aim of this study was to assess the potential of the mono- and di-isopropyl esters of glutathione (MIPE and DIPE respectively) as cytoprotectants in the human upper respiratory tract cell lines BEAS-2B and RPMI 2650. 2. The optimal concentrations for cytoprotection were shown to be 1.0 mg/ml for both DIPE and MIPE. Both compounds were found to protect cells by pretreatment, slightly less protection was observed in cells simultaneously exposed to sulphur mustard. The greatest protection was shown where MIPE or DIPE were in in situ at the time of exposure to HD. The optimum pre-treatment times were found to be 1 h for MIPE and 2 h for DIPE. Limited protection of cells treated with MIPE or DIPE immediately following HD exposure was also demonstrated. No protection was observed if MIPE or DIPE were not administered immediately following HD exposure. 3. Results suggest that MIPE and DIPE may be effective treatments for exposure to HD by inhalation. PMID- 9726534 TI - Subchronic effects of ochratoxin A on young adult rat brain and partial prevention by aspartame, a sweetener. AB - 1. Ochratoxin A (OTA) is a mycotoxin produced by several fungi, especially Aspergillus and Penicillium species. Many food and foodstuffs can be contaminated by ochratoxin A, which is consequently found in blood of animals and humans. 2. The distribution into the brain of young adult rats fed OTA for 1 to 6 weeks and some consequences have been investigated in the present study. 3. Our results on rats given OTA (289 microg/kg/48 h) indicated that OTA accumulated in the whole brain as function of time according to a regression curve, Y=-8.723 a+16.72 with a correlation coefficient of r=0.989, where Y-axis is the OTA concentration in ng/g of brain and X-axis is the duration of the treatment in weeks. The brain OTA contents was 11.95 +/- 2.2, 23.89 +/- 4.4, 39.9 +/- 4.5, 50.3 +/- 7.3, 78.8 +/- 6.3, 94 +/- 16 ng/g of brain in the mycotoxin-treated animals for respectively 1, 2, 3, 4, 5 and 6-weeks treatment. OTA induced modifications of free amino-acid concentrations in the brain, mainly, Tyrosine (Tyr) and phenylalanine (Phe). Tyr decreased significantly as compared to control (p < 0.05). Phe increased significantly as compared to control (p < 0.05). 4. Aspartame, (25 mg/kg/48 h) a structural analogue of OTA largely modified the distribution and prevented the accumulation of OTA in the brain since the respective brain OTA contents decreased respectively to 9.6 +/- 7.9, 19.2 +/- 3.0, 26.8 +/- 4.2, 19.7 +/- 1.9, 13.7 /- 5.6 and 11.0 +/- 6.0 ng/g of tissue, for the same duration of treatment. It also prevented the modifications of Tyr and Phe levels. 5. The histological investigations showed several necrotic cells with pyknotic nucleus, detected in OTA treated animals with higher frequency as compared to the controls and Aspartame treated ones. Aspartame appeared to significantly prevent this nuclear effect as well, the meaning of which is discussed. PMID- 9726536 TI - Castration prevents calcium channel blocker-induced gingival hyperplasia in beagle dogs. AB - 1. The purpose of this study was to investigate testosterone's role on the calcium channel antagonist oxodipine-inducing gingival hyperplasia in a dog model. 2. Two experiments were conducted using castrated and intact male dogs. Oxodipine was administered orally for 90 days, at a dose of 24 mg/kg/day. In the first experiment, the occurrence of gingival hyperplasia was evaluated. In the second, the gingival index (GI) and gingival hyperplasia index (GHI) were recorded and correlated with serum levels of testosterone. 3. A significant positive correlation between GI, GHI and plasma testosterone was noted. Castrated dogs were injected with testosterone, 4 months after the start of oxodipine treatment, while in the non-castrated dogs, administration of oxodipine was stopped. Castration correlated with lack of GH, while testosterone injection to the same dogs was associated with an increase of GI and GHI. 4. Since it is known that testosterone receptors are present in the gingiva, it is proposed that oxodipine-induced gingival hyperplasia could be mediated by the calcium channel blocker on plasma testosterone levels. PMID- 9726537 TI - Unique cerebral dysfunction following triphenyltin acetate poisoning. AB - 1. In animal studies, TPTA was found to be neurotoxic. In humans, variable CNS pictures have been described with or without significant EEG findings. Brain CT does not usually reveal any abnormalities. 2. Our patient presented with intermittent unique spontaneous involuntary movement of hands, facial twitching, silly smile and crying. Diplopia, drowsiness, giddiness, vertigo, bidirectional nystagmus, impairment of calculation ability, as well as disorientation to time, people and place also developed. EEG showed mild cortical dysfunction without seizures. MRI and Tc-99m HMPAO brain SPECT revealed no significant findings. TPTA may cause cellular dysfunction of brain without structural damage, which results in variable CNS clinical presentations. 3. Nadir of leucopenia was noted on the sixth day after consumption of TPTA. Liver impairment occurred on the ninth day. Borderline demyelinated neuropathy developed on the fifty-third day. CNS abnormalities, delayed peripheral neuropathy, hepatitis and leucopenia deserve monitoring for a prolonged period, even when the victim initially presents with GI upset only after consumption of TPTA. PMID- 9726538 TI - A dose-dependent delayed hypersensitivity reaction to acetaminophen after repeated acetaminophen intoxications. AB - We report a case of a 29-year-old woman with a borderline personality disorder who presented with intentional substantial acetaminophen (paracetamol) overdosage on nine occasions during a period of 21 months. In most cases, the patient presented at the hospital within 4 h after ingestion and was treated with gastric lavage, activated charcoal, laxatives and intravenous N-acetylcysteine. During the sixth overdosage the patient developed a rash on her chest and shoulders which was considered an anaphylactoid reaction to N-acetylcysteine. Therefore she was treated with oral methionine subsequently, but developed the rash again. The rash was then ascribed to the repeated high-doses of acetaminophen and treatment with N-acetylcysteine was reinstituted. This case shows that when an anaphylactoid reaction occurs after an acetaminophen overdose and treatment with N-acetylcysteine, acetaminophen must also be taken into account as the cause of the anaphylactoid reaction before effective therapy with N-acetylcysteine is withheld. PMID- 9726539 TI - Neurotrophins, neurones and peripheral nerve regeneration. AB - Successful peripheral nerve regeneration requires optimal conditions both in the macro-environment and micro-environment. Many methods have been used to improve the macro-environment for the regenerating nerve. However, much less is known about the micro-environment, and in particular the complex neurochemical interactions involved. Several neurotrophic factors have been shown to play an essential trophic role in the development, maintenance and regulation of neuronal function. These include nerve growth factor (NGF) and several recently identified members of the NGF family, namely brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), neurotrophin-4/5 (NT-4/5) and neurotrophin-6 (NT-6). In this review we summarize recent studies of the effects of these neurotrophins on neurones, especially their effects on motor neurones and their axonal outgrowth. We discuss prospects for the future and point out what remains to be understood about the role of neurotrophins to enhance peripheral nerve regeneration. PMID- 9726540 TI - Ulnar lengthening in juvenile chronic arthritis. AB - Ten wrists in eight children with severe destructive changes in the wrist due to juvenile chronic arthritis underwent distraction lengthening of the ulna between 1983 and 1996 in an attempt to restore wrist alignment, which had not been preserved by previous splinting. Eight wrists in six patients (four female and two male) were evaluated. The average age at the time of operation was 13.8 years (range, 12-15.5). The average follow-up period was 70 months (range, 12-152). The average ulna minus deformity was between 8 and 9 mm. It was found that ulnar lengthening can be done safely and the procedure seems to stabilize the carpus, is likely to improve appearance, may improve function and in the majority of cases does eliminate the use of an external splint. Results seem to be stable for at least 3 to 5 years. PMID- 9726541 TI - Correction of soft tissue contractures of the wrist using the Ilizarov technique. AB - The Ilizarov technique can allow new histiogenesis of soft tissue as well as bone. Three cases of wrist contracture successfully treated by this method are described. One patient had transient superficial pin site infections. There were no other complications. At 2 years follow-up there was no recurrence of contractures. PMID- 9726542 TI - Distraction and microvascular epiphysis transfer for radial club hand. AB - This paper presents a different technique of treatment for Bayne type IV radial club hand using a microvascular joint transfer in order to reconstruct the absent half of the wrist joint, aiming for better movement and stability at the wrist joint with preservation of longitudinal growth. The method uses preoperative soft tissue distraction to obtain proper alignment of the hand on the ulna before the second metatarsophalangeal joint with the whole metatarsal bone is transplanted. The treatment takes about 4 months and the optimum period for surgery is during the second year of life. Pollicization is added later in the normal manner. The new technique has been used in 12 cases by the author since 1987 and the results of the first nine cases are reported with a mean follow-up of 6 years. This technique appears to be promising but is demanding because of the microvascular joint transplantation at an early age. PMID- 9726543 TI - Lengthening of a one-bone forearm. A sequel of neonatal osteomyelitis. AB - An 8-year-old girl presented with marked shortening of the right forearm due to destruction of both the radius and ulna secondary to neonatal osteomyelitis. A one-bone forearm operation was performed to achieve a stable forearm. Two years later, the one-bone forearm was lengthened for 6 months by callus distraction (callotasis) achieving 12 cm of extra length. The patient was last followed up at the age of 16. The appearance and functional outcome of the right upper limb had been improved and she was independent in all activities of daily living. PMID- 9726544 TI - Reconstruction of fingertip amputations by partial composite toe transfer with short vascular pedicle. AB - Five patients under 15 years-of-age, with a fingertip amputation through the nail plate, were treated with a custom-made partial toe transfer. Two of the patients had had previous attempts at surgical reconstruction using either a local flap or replantation. Delay between initial injury and reconstruction ranged from 2 to 60 days. In all cases the flap was harvested from the second toe. This "custom-made" compound transfer included the exact amount of pulp, nail bed and bone required for reconstruction. All flaps were harvested on a short vascular pedicle, with anastomoses performed at a digital level on the recipient site. Good to excellent cosmetic results were obtained in all cases, with a nearly normal-looking fingertip. Duration of hospital stay ranged from 4 to 7 days. We recommend this technique for treatment of distal amputation close to the proximal nail fold, in young individuals. PMID- 9726545 TI - Ring avulsion injuries managed with homodigital and heterodigital Venous Island Conduit (VIC) flaps. AB - Ring avulsion injury frequently results in vascular insufficiency (venous or arterial) and soft tissue injury. We report four cases requiring revascularization where venous congestion and dorsal skin cover were achieved using a composite pedicled venous flow-through flap. We have termed this a Venous Island Conduit (VIC) Flap. Two types of flaps, homodigital and heterodigital, were used, depending on the severity of the injury. The techniques and results are discussed. Venous island conduit flaps are ideally suited to the management of ring avulsion injury and have several advantages over the alternatives. PMID- 9726546 TI - The subcutaneous turn-over flap in the treatment of difficult wounds of the digits. PMID- 9726547 TI - The radial forearm fascial flap as a salvage procedure for chronic tuberculosis of the hand and wrist. AB - A patient with tuberculous infection of the hand and wrist developed a recurrent draining wound of the right forearm. After recurrent failure of surgical debridement and wound closure under antituberculous therapy, wound closure was established by means of a radial forearm fascial flap with an excellent functional and cosmetic result. Extra-pulmonary tuberculosis must be kept in mind in the diagnosis of slowly growing tumours and chronic wounds in the upper extremity. PMID- 9726548 TI - A 10-year review of high-pressure injection injuries to the hand. AB - High-pressure injection injuries to the hand are uncommon, but often result in poor outcome or even amputation. We report a review of the 28 cases treated surgically in our department over the last 10 years and have examined the factors leading to increased morbidity. The severity of these injuries was related to the nature of the injected material, involvement of the tendon sheath and proximal spread of the injected substance. All cases in which the digit was noted to be poorly perfused from the outset resulted in amputation. We conclude that early amputation should be considered in cases in which the affected digit is initially cool or poorly perfused. PMID- 9726549 TI - Hand lacerations. An audit of clinical examination. AB - Over a 6-month period 147 consecutive deep hand lacerations referred to a plastic surgery department were explored in 136 adult patients. At operation 121 complete tendon divisions, and 72 nerve injuries were found. Accident and emergency (A&E) officers diagnosed only 64% of tendon divisions compared with 84% for the admitting hand surgeons. Nerve injuries were more accurately diagnosed, 87% by A&E officers compared with 94% by hand surgeons. These findings support the view that significant palmar hand lacerations should be referred for a hand surgery opinion. PMID- 9726550 TI - Outcome following the rehabilitation of hand trauma patients. The importance of a subjective functional assessment. AB - Objective measures of hand function have been used to assess the outcome of rehabilitation following trauma. However, subjective assessments of function have been avoided by clinicians due to the difficulty in proving their validity and reliability. We have developed a subjective hand function scoring system (HFS), based on an activities of daily living assessment, which is used in planning and monitoring progress through rehabilitation. The HFS for 64 traumatic hand injuries were assessed on admission and discharge, and a significant improvement was found. There was a positive correlation between the HFS on admission and both the severity of injury, and the length of time off work. This scoring system is not validated, but this study illustrates the use of subjective functional scoring systems in the planning, delivery and evaluation of rehabilitation. PMID- 9726551 TI - Transthecal digital nerve block. An anatomical appraisal. AB - Injection studies using methylene blue and latex were used in 60 digits from 40 cadavers to study how anaesthetic fluid injected into the flexor tendon sheath might spread around the proximal part of the finger. The injected solution escaped from the flexor tendon sheath around the vincular vessels which are present near the base and head of the proximal phalanx. Outside the digital canal, the dye flowed smoothly through the perivascular loose areolar tissue and spread alongside the main digital vessels and nerves and their palmar and dorsal branches. PMID- 9726552 TI - A method to locate the radial digital nerve of the index finger. AB - The radial digital nerve of the index finger is susceptible to injury during penetrating trauma or elective release of the A1 pulley. The intersection of a line drawn down the midline of the index finger and the proximal palmar crease identifies the location of the radial digital nerve. This method of identifying the topography of the nerve should assist the surgeon in determining the likelihood of injury after penetrating trauma, and preventing injury during elective procedures. PMID- 9726553 TI - Defining a normal finger web in children. AB - We calculated the interdigital web ratios for children between the ages of 5 and 6 and 10 and 11 years. We compared these ratios with those seen in adults and conclude that there is a change in finger web morphology with age. We found that a photocopier provided images of similar quality to conventional photography. PMID- 9726554 TI - The reliability of physical examination for carpal tunnel syndrome. AB - The goal of this study was to determine the interobserver and intraobserver reliability of static and moving two-point discrimination, Semmes-Weinstein monofilament testing, Tinel's test, manual motor testing of abductor pollicis brevis, vibration and Phalen's test in the diagnosis of carpal tunnel syndrome. Twelve patients with suspected carpal tunnel syndrome were examined in an outpatient setting. The interobserver reliability was satisfactory for all tests except for Semmes-Weinstein monofilament testing. Intraobserver reliability was also satisfactory for all tests. Static two point discrimination had higher reliability than moving two-point discrimination. Seven tests for the diagnosis of carpal tunnel syndrome were reliable in the hands of skilled health care professionals. Hand surgeons and hand therapists examined patients more reliably than occupational health workers. PMID- 9726555 TI - Chronic post-traumatic radial instability of the metacarpophalangeal joint of the finger. Long-term results of ligament reconstruction. AB - Chronic painful post-traumatic instability of the radial collateral ligament complex of the metacarpophalangeal joint of a finger was treated by tendon graft reconstruction in 24 patients. Seventeen patients (20 joints) were available for a retrospective study at a mean follow-up time of 105 months. Eighty percent of the joints showed excellent or good results, with relief of pain, return of adequate stability, a near normal range of motion and absence of degenerative changes. PMID- 9726556 TI - The importance of the distal radioulnar joint in distal radial fractures. AB - In a prospective study we evaluated the results of 272 distal radial fractures by their involvement of the distal radioulnar joint. Impaired function following altered anatomy at the distal radius can be explained by dysfunction of the distal radioulnar joint. Ulnar styloid avulsions contribute to a poorer result because of their effect on distal radioulnar joint function. PMID- 9726557 TI - The clinical importance of carpal instabilities following distal radial fractures. AB - A prospective study was undertaken to determine the clinical importance of the different carpal instabilities following dorsally displaced distal radial fractures (Colles' type). All patients were followed for 1 year and a Cooney score and X-ray evaluation were done. Nine different carpal instabilities were evaluated. Only dissociative DISI and ulnar translocation showed significant clinical differences at 1 year follow-up. It is therefore recommended that a dissociative DISI, usually caused by scapholunate dissociation, should be treated by percutaneous pinning at the time of the initial treatment. PMID- 9726559 TI - Morphology and distribution of nerve endings in the human triangular fibrocartilage complex. AB - We studied the morphology and distribution of nerve endings in the human triangular fibrocartilage complex using both silver staining and immunohistochemical staining using a protein specific to nerve fibres. Free nerve endings were found in the ulnar side of the triangular fibrocartilage complex, especially in the ulnar collateral ligament, meniscus homologue and the adjacent collagen fibre area of the peripheral part of the ulnar side of the articular disc. Meissner's and Krause's corpuscles were observed in the ulnar collateral ligament and meniscus homologue. The fact that free nerve endings were observed in the meniscus homologue and adjacent collagen fibre area of the peripheral part of the ulnar side of the articular disc suggests that this disc may be a source of wrist pain. The presence of nerve end bulbs in the triangular fibrocartilage complex also suggests a possible role for corpuscles as mechanoreceptors. PMID- 9726558 TI - Distal ulnar tumours. Results of management by en bloc resection in nine patients and review of the literature. AB - A variety of reconstructive procedures have been suggested for stabilizing the ulnar shaft following resection of the distal ulna for tumour. We present the results of a series of nine distal ulnar tumour resections in which four different stabilization techniques were employed. We based our results on an evaluation of function, pain, motion, strength and instability. We obtained good or excellent results in seven patients treated with a soft tissue stabilization of the ulnar stump. One patient did not undergo any stabilization procedure and scored fair in our system. A further patient who required a radiocarpal arthrodesis also had an inferior result. These results suggest that soft tissue stabilization of the ulnar stump should be performed whenever possible. PMID- 9726560 TI - Psychomotor development in children with triphalangeal thumbs. A preliminary study. AB - In order to explore the influence of an isolated congenital hand malformation on psychomotor development, we performed an exploratory, observational study on 18 children with triphalangeal thumbs. The investigative procedure consisted of a hand function examination, a semi-structured interview with the mother about the development of the child, the so-called "Hand test", and the "Child Behaviour Check List". Our observations suggest specific developmental difficulties in fine motor skills and language development, but the children showed no signs of behavioural psychopathology. PMID- 9726561 TI - A large family with type IV radial polydactyly. AB - This study examines one of the largest pedigrees with radial polydactyly type IV (uncomplicated polysyndactyly) comprising a total of 69 individuals, of whom 26 have been affected over six generations. Typical manifestations of the pedigree were bilateral radial and ulnar digital duplications, as well as syndactyly between the middle and ring fingers and the second and third toes. There was no craniofacial anomaly in any of the 17 cases examined physically. This observation suggests that radial polydactyly type IV and Greig craniofacial-synostosis syndrome with similar digital manifestations are clinically-distinct entities. PMID- 9726562 TI - Classification of the mirror hand-multiple hand spectrum. AB - A rare variant of mirror hand is described. The hand had eight fingers and the forearm contained an ulna and a hypoplastic radius. A classification of the mirror hand-multiple hand spectrum is offered and its embryology discussed. PMID- 9726563 TI - An unusual carpal coalition associated with fifth ray anomalies in the hand. AB - I report the case of a 15-year-old white girl with an asymptomatic scapholunotriquetral fusion associated with a capitometacarpal coalition in the right wrist. She had additional anomalies of the fifth ray including shortening and ulnar deviation of the fifth metacarpal, brachydactyly of the little finger, fusion of the fourth and fifth metacarpals and absence of the ulnar styloid. PMID- 9726564 TI - Is occupation an aetiological factor in the development of trigger finger? AB - We investigated the occupation histories of 178 patients with idiopathic trigger finger. When compared with the 1991 Census data, the distribution of their occupations was not significantly different from the local general population. It is concluded that the vast majority of trigger fingers develop for reasons other than occupation. PMID- 9726565 TI - Trigger finger in a child as a complication of interphalangeal dislocation. PMID- 9726566 TI - A rare combined fracture and ligamentous injury of the thumb. AB - Bennett's fracture of the base of the metacarpal of the thumb and avulsion of the ulnar collateral ligament of the thumb are both relatively common injuries. It is, however, rare for the two lesions to occur simultaneously and we report such a case. PMID- 9726567 TI - Atraumatic dislocation of the trapeziometacarpal joint secondary to osteoarthritis. AB - We report a case of atraumatic dislocation of the trapeziometacarpal joint secondary to osteoarthritis. An attritional rupture of the anterior oblique carpometacarpal ligament is thought to have occurred, allowing complete dislocation which has led to a reduction in the patient's pain. PMID- 9726568 TI - Palmar metacarpophalangeal joint dislocation. AB - Palmar dislocations of the long finger metacarpophalangeal joint are extremely rare and easily missed at the first clinical examination. We describe a palmar metacarpophalangeal dislocation of the long finger following a hyperflexion injury. The presentation, aetiology and treatment are discussed. PMID- 9726569 TI - Rupture of the tendon of flexor digitorum profundus in association with an endchondroma of the terminal phalanx. PMID- 9726570 TI - Avascular necrosis of the trapezoid bone. AB - A case of idiopathic avascular necrosis of the right trapezoid is presented. The aetiology was not clear. Treatment consisted of bone curettage, autologous bone graft and revascularization with a dorsal metacarpal artery. PMID- 9726571 TI - Avascular necrosis of the metacarpal head. AB - A case of avascular non-traumatic necrosis of the metacarpal head in a 36-year old woman is reported. Treatment was by curettage of the necrotic bone and packing with cancellous grafts. PMID- 9726572 TI - A false aneurysm in the hand of a child. AB - This is a case report of a 4-year-old child presenting with a false aneurysm of a common digital artery. Failure to diagnose it led to incorrect treatment which was followed by rupture of the aneurysm and life-threatening haemorrhage. PMID- 9726573 TI - Delayed rupture of an extensor digitorum tendon following repeated attempts at intravenous cannulation. AB - A case of delayed extensor tendon rupture is reported. This followed repeated attempts at intravenous cannulation 16 months previously. The differential diagnosis and treatment are discussed. PMID- 9726575 TI - Carpal tunnel decompression under local anaesthetic and tourniquet control. PMID- 9726574 TI - Tendon interposition in a juxta-epiphyseal fracture of the proximal phalanx. AB - Juxta-epiphyseal fractures of the proximal phalanx are relatively common. A case report and literature review calls attention to the possibility of entrapment of either a flexor or extensor tendon in this type of fracture. PMID- 9726576 TI - Carpal tunnel decompression under local anaesthetic and tourniquet control. PMID- 9726577 TI - The use of positron emission tomography scanning in occult and recurrent head and neck cancer. PMID- 9726578 TI - Dysembryoplastic neuroepithelial tumour. AB - We report here a classic case of dysembryoplastic neuroepithelial tumour, which was situated in the left frontal lobe of a 13-year-old boy who presented with intractable partial complex seizures since 3 years of age. This tumour entity was first described in 1988, and has been incorporated in the 2nd edition (1993) of the WHO histological typing of CNS tumours. Fewer than 100 cases have been reported from all over the world. To our knowledge, this is the first report of this tumour from India. It is important to recognise this entity since surgery is the only means of cure and radio- or chemotherapy is unwarranted. PMID- 9726579 TI - Perioperative specific management of blood volume loss in craniosynostosis surgery. AB - Accurate assessment and replacement of blood loss and fluid-electrolyte deficit during craniosynostosis repair is difficult owing to patient size and the diversity of surgical technique. Forty-three patients undergoing primary craniosynostosis repair over a 10-year period were studied retrospectively to determine blood loss and fluid deficit and to assess blood transfusion practices during both intraoperative and postoperative periods. Blood loss was calculated on the basis of estimated red cell mass (ERCM) and fluid-electrolyte imbalance was investigated with blood samplings. Blood transfusion was considered appropriate if the postoperative or posttransfusion ERCM was within 12% of the preoperative value. Estimated fluid requirement (EFR) was used in 4 ml kg(-1) h( 1) except for neonates. Intraoperatively, 80% of all patients were appropriately managed with respect to blood transfusion and EFR. Postoperatively only 20% of the patients receiving transfusions were transfused appropriately. In 23.3% of these patients (10/43) unexpected respiratory distress developed immediately after their recovery from the anesthesia. With the measurement of estimated blood volume and allowable blood loss, appropriate transfusion could be achieved for the successful treatment of the primary craniosynostosis. PMID- 9726581 TI - Medullomyoblastoma. A rare cerebellar tumour in children. AB - Seven patients between the ages of 3 and 24 years were admitted to our hospital in the last 28 years who had a histological diagnosis of medullomyoblastoma. These patients presented with classic symptoms of a posterior fossa midline mass associated with evidence of raised ICP. A CT scan in each patient revealed a uniformly high-attenuating tumour in the posterior fossa with gross hydrocephalus. In all seven patients a ventriculoperitoneal shunt was placed prior to definitive surgery. Radical tumour excision was carried out in all cases 3-5 days after CSF diversion. The histological diagnosis was made on H&E-stained slides. In two cases each, the tumour tissue was subjected to electron microscopy and immunohistochemical studies. Six of the seven patients survived the operation. One patient died 21 days after surgery as a result of shunt block and shunt infection. All surviving patients received cranial and spinal radiation 2-4 weeks after surgery, and also chemotherapy. The cranial radiation dose ranged from 4500 to 5000 rad, while the spinal radiation dose was limited to 1500 rad. Patients were followed up carefully. Three patients died within 6 months, and the remaining three between 2.5 and 3 years after surgery. None of the patients in our study survived longer than 3 years. One patient had developed paraplegia. This study highlights the details of an uncommon entity and reports the largest collection of such cases in the literature. PMID- 9726580 TI - Epidemiology and prognosis in children treated for intracranial tumours in Denmark 1960-1984. AB - A total of 911 Danish children under 15 years of age were treated for an intracranial tumour in the 25-five year period 1960-1984. All cases were followed up to the end of 1994 or to emigration or death if one of these came sooner. The mean annual incidence was 32.5 per million children with a slight increase over the 25 years. The male/female ratio was 1.15 and close to the M/F ratio for the entire Danish population of children. Of the tumours, 46% were located in the supratentorial and 54% in the infratentorial compartment, and 94% were verified histologically. In order of frequency the most common types were astrocytomas (all grades, 35%), medulloblastomas (20%), ependymomas (14%), and craniopharyngiomas (5%). Total removal of the tumour was performed in 277 and partial removal, including biopsy, in 490 children. In 57 patients a shunt operation only was performed, and 87 children did not have an operation or died before the correct diagnosis was established. Radiotherapy was administered in 55%. The outcome depended on extent of removal, radiation, location and histology of the tumour. Most (784 or 86%) of the children survived more than 1 month after diagnosis or operation, and 353 children (39% of the whole series, 47% of those alive more than 1 month after diagnosis) were alive at follow-up. Of the survivors 29% had a tumour in the supratentorial midline, 26% one in the lateral part of the supratentorial area, 31% a cerebellar tumour and 13% a IV ventricle tumour. It was possible for 66% of the survivors with supratentorial and 90% of those with infratentorial tumours to lead a normal life. The long-term prognosis was especially good for children with cerebellar and supratentorial astrocytomas and optic chiasma tumours. Children with juvenile cerebellar astrocytoma had the best prognosis: 90% were alive at the end of the follow-up period, as against 20% of those with medulloblastoma and 6% of those with glioblastoma. A comparison of the data from the present series and from a similar Danish series of intracranial tumours in 533 children seen in the years 1935-1959 shows no significant differences in location or histology, a slight increase in annual incidence, and improved survival rates during the 50 years in question. PMID- 9726582 TI - Chemotherapy for spinal cord astrocytoma: can natural history be modified? AB - Standard treatment of spinal cord astrocytomas is based upon surgery, followed by radiotherapy when resection is incomplete or when histology is of high grade. Owing to the major consequences of radiotherapy on the spine in childhood, alternative therapies must be explored. The potential role of chemotherapy in the management of spinal cord astrocytoma remains to be defined. Two patients are described. The first was a 19-month-old child with an anaplastic astrocytoma of the cervical spinal cord that progressed rapidly after initial partial resection. Chemotherapy was begun according to the UKCCSG Baby Brain Protocol, with marked clinical improvement. Reassessment by MRI at 4 months showed improvement, and at the end of treatment no evaluable disease remained. The second was a 4-year-old child with a recurrent low-grade astrocytoma. Chemotherapy according to the SIOP Protocol for Low Grade Gliomas was administered for 3 months, after which marked tumour regression was seen, with neurological recovery. These patients demonstrate the potential value and low morbidity of chemotherapy in spinal cord astrocytoma. The management of this rare tumour is discussed. PMID- 9726583 TI - Computer simulation of a neurosurgical operation: craniotomy for hypothalamic hamartoma. AB - Although magnetic resonance imaging has revolutionised the management of intracranial lesions with improved visualisation of anatomical structures, it only produces two-dimensional images, from which the clinician has to extrapolate a three-dimensional interpretation. Several approaches can be used to create 3D images; the discipline of image segmentation has encompassed a number of these techniques. Such techniques allow the clinician to delineate areas of interest. The resulting computer-generated outlines can be reconstructed in a three dimensional arrangement. Although a plethora of "generic" segmentation techniques exist, we have developed a refined form, dependent on general and particular properties of the anatomical structures under investigation. High-contrast structures such as the ventricles and external surface of the head are found by using a localised adaptive thresholding technique. Less definable structures, with poor or nonexistent signal change across neighbouring structures, such as brain stem or pituitary, are found by applying an "energy minimisation"-based technique. To demonstrate the techniques we used the example of an 8-year-old boy with uncontrolled gelastic seizures due to a hypothalamic hamartoma, who is being considered for surgery. We were able to demonstrate the anatomical relationships between the hypothalamic hamartoma and adjacent structures such as optic chiasm, brain stem and ventricular system. We were subsequently able to create a video, reproducing the stages of craniotomy for excision of this tumour. By creating true 3D objects, we were able at any stage of the simulation to visualise structures situated contralaterally to the approaching surgical dissector. These 3D representations of the structures can be either invisible or opaque, in order to afford 3D localisation as the "virtual" surgical dissection proceeds. The clinical application of such techniques will enable surgeons to improve their understanding of anatomical relations of intracranial lesions and has obvious implications in image-guided surgery. PMID- 9726584 TI - The role of somatosensory evoked potentials in spinal dysraphism--do they have a prognostic significance? AB - Somatosensory evoked potentials (SEP) are not routinely used in spinal dysraphism. In this study 38 patients (29 children and 9 adults) with spinal dysraphism were prospectively studied with the objective of evaluating whether SEPs were a prognostic tool that could be used to predict clinical improvement after repair of a spinal dysraphic lesion. For all patients, preoperative SEP and postoperative SEP (within 1 week of operation) were recorded. Fifteen of these patients also had follow-up postoperative SEP recordings taken within 3 months of operation. A clinical examination was performed at the time of each SEP. Thirty patients had tethered cord, 12 had diastematomyelia and 15 had intra- and/or extradural tumours, which included lipomas and dermoid and epidermoid tumours. Twenty-one children and all adults had abnormal preoperative SEPs. Sixteen children and 4 adults had improved SEPs postoperatively. All these children and 2 of the 4 adults also experienced clinical improvement. Improvement in SEPs preceded clinical improvement in 12/20 patients. We observe that SEPs have a good prognostic value. PMID- 9726585 TI - Pneumocephalus after shunting for hydrocephalus. AB - A case of tension pneumocephalus that occurred after ventriculoperitoneal shunting is presented. We have reviewed 12 cases of pneumocephalus in association with ventriculoperitoneal shunt placement. This phenomenon occurs when air is forced through the shunt or enters through the cranial base because of: iatrogenic postsurgical connection, congenital fistula, trauma, or thinning of the cranial base. Ways of preventing and treating this problem are outlined. PMID- 9726586 TI - Unusual presentation of a sinonasal carcinoma mimicking an aneurysm rupture. AB - Although the association of subarachnoid hemorrhage (SAH) and tumoral lesions in adult is well known, hemorrhage from a sinonasal carcinoma extending to the intracranial cavity is exceedingly rare. In this paper, the authors report on a 12-year-old girl who presented with SAH caused by a sinonasal carcinoma located in the anterior skull base area. To our knowledge, this is the first report of a sinonasal carcinoma concomitant with SAH. PMID- 9726587 TI - A case of Rathke's cleft cyst with apoplexy. AB - We report a case of Rathke's cleft cyst associated with cholesterin granuloma in an 8-year-old girl with apoplexy. She was admitted to our hospital in April 1996 because of repeated headache and deep ophthalmic pain, without any visual disturbance. Computed tomography (CT) of the pituitary demonstrated an intrasellar isodense mass extending to the suprasellar cistern. Magnetic resonance imaging (MRI) showed a high-intensity mass on both T1- and T2-weighted images. The preoperative diagnosis of this lesion was Rathke's cleft cyst associated with a craniopharyngioma and/or hemorrhage. Transsphenoidal microsurgery was performed, and a bloody coffee-like serous and mucinous yellowish substance was evacuated. Curettage of the wall removed the yellowish hard mass and soft membranous tissue. Histological examination of this tumor revealed a Rathke's cleft cyst with cholesterin granuloma. PMID- 9726588 TI - Obesity, diabetes and the central nervous system. PMID- 9726589 TI - Relation of fibre intake to HbA1c and the prevalence of severe ketoacidosis and severe hypoglycaemia. EURODIAB IDDM Complications Study Group. AB - The effect of dietary fibre intake on glycaemic control is still controversial. This study analysed the intake of natural dietary fibre in patients with Type I diabetes mellitus enrolled in the EURODIAB IDDM Complications Study to determine any associations with HbA1c levels and with the prevalence of severe ketoacidosis or severe hypoglycaemia. Dietary intake was assessed by a 3-day dietary record. The relation between intake of fibre (total, soluble and insoluble) and HbA1c was examined in 2065 people with Type I diabetes. Associations with severe ketoacidosis (requiring admission to hospital) and severe hypoglycaemia (requiring the help of another person) were analysed in 2687 people with Type I diabetes. Total fibre intake (g/day) was inversely related to HbA1c (p = 0.02), independently of carbohydrate intake, total energy intake and other factors regarding lifestyle and diabetes management. Severe ketoacidosis risk fell significantly with higher fibre intake (p = 0.002), with an adjusted odds ratio of 0.48 (95 % confidence interval 0.27 to 0.84) in the highest quartile ( > or = 23.0 g fibre/day) compared with the lowest quartile ( < or = 13.7 g fibre/day). The occurrence of severe hypoglycaemia was not related to fibre intake. Beneficial effects of fibre on HbA1c and the risk of severe ketoacidosis were particularly pronounced in patients from southern European centres. This study shows that higher fibre intake is independently related to a reduction in HbA1c levels in European people with Type I diabetes. Furthermore, increased fibre intake may reduce the risk of severe ketoacidosis. These beneficial effects were already observed for fibre intake within the range commonly consumed by people with Type I diabetes. PMID- 9726590 TI - Immunological heterogeneity in type I diabetes: presence of distinct autoantibody patterns in patients with acute onset and slowly progressive disease. AB - Type I diabetes mellitus may represent a heterogeneous disorder with a distinct pathogenesis in patients with young and adult onset of the disease. To investigate whether serological markers directed to different autoantigens have the potential to distinguish acute onset from slowly progressive Type I diabetes we analysed antibodies to tyrosine phosphatases IA-2/ICA512 (IA-2A) and IA 2beta/phogrin (IA2betaA), antibodies to GAD65 (GADA) and cytoplasmic islet cell antibodies (ICA) in a non-selected group of diabetic patients clinically classified as having Type I or Type II diabetes at diagnosis. Both IA-2A and IA 2betaBA were found to be positively associated with onset before the age of 20 years and the presentation of classical features of Type I diabetes. In Type I diabetes 56 % (112/200) of patients were positive for IA-2A and 38 % (76/200) for IA-2betaA. In contrast, only 1 of 785 (0.1 %) patients with Type II diabetes had IA-2A and all of them were negative for IA-2betaA (p < 0.001). Among the patients with Type II diabetes 7.6% (n = 60) were ICA positive and 2.8% (n = 22) had GADA suggesting the presence of slowly progressive Type I diabetes. GADA were found in 8 of 60 (13.3 %) ICA positive subjects which was lower than the percentage detected in patients with acute onset of diabetes (115/157 73.2%) (p < 0.001). Blocking of double antibody positive sera showed that only 3 of 8 (37.5 %) patients with slowly progressive diabetes had ICA restricted to GAD or IA-2 whereas ICA were completely inhibited in 12 of 20 (60.0 %) patients with Type I diabetes. Among 193 patients with Type II diabetes available for follow-up, 35 % of ICA positives, 58 % of GADA positives and 60 % of those positive for both markers required insulin by 3 years. However, using strict criteria for the switch to insulin treatment the corresponding sensitivity of each marker was only low (9%, 10% and 5%). We show that clinical subtypes of Type I diabetes are associated with distinct humoral autoimmunity. IA-2A and GADA were associated with classical features of Type I diabetes whereas GADA and an uncharacterized ICA subspecificity indicate slowly progressive disease. PMID- 9726591 TI - Prospective assessment of severe hypoglycaemia in diabetic children and adolescents with impaired and normal awareness of hypoglycaemia. AB - To establish whether impaired hypoglycaemic awareness is associated with increased rate of severe hypoglycaemia and to assess clinical predictors of severe episodes without warning symptoms a prospective study of 130 insulin dependent diabetic children and adolescents was undertaken for 1 year. Using a structured questionnaire, 48 patients reported impaired awareness and 82 reported normal awareness of hypoglycaemia at baseline of the study. The two groups did not differ regarding clinical and metabolic characteristics. Episodes of severe hypoglycaemia were recorded for 1 year. The rate of severe hypoglycaemia was higher in the group with impaired awareness than in the group with normal awareness (p < 0.0001). Of the severe hypoglycaemic episodes, 34.0% developed without warning symptoms. Patients with impaired awareness experienced more severe episodes without warning symptoms than those with normal awareness (p = 0.0054). Severe hypoglycaemia occurred more frequently in patients with impaired awareness aged 6 years and less (p = 0.0041) than in older counterparts. Impaired awareness reported at baseline [adjusted odds ratio (OR): 5.8; p =0.0021], age 6 years or less (3.4; p = 0.0121), previous severe episode (4.8; p = 0.0043) and more than 5 % of home blood glucose readings 3.3 mmol/l or less in the preceding month (4.2; p = 0.0211) proved to be independently predictive of severe hypoglycaemic events without warning symptoms. In conclusion, impaired hypoglycaemic awareness is associated with an increased rate of severe hypoglycaemia in diabetic children and adolescents. One third of severe episodes developed without warning symptoms. Impaired awareness, young age and recent biochemical or severe hypoglycaemias are independent risk factors for such episodes. Avoidance of hypoglycaemia should be a priority in preschool children with diabetes. PMID- 9726592 TI - Increasing prevalence of Type II diabetes in American Indian children. AB - Until recently, Type II diabetes was considered rare in children. The disease is, however, increasing among children in populations with high rates of Type II diabetes in adults. The prevalence of Type II diabetes was determined in 5274 Pima Indian children between 1967 and 1996 in three 10-year time periods, for age groups 5-9, 10-14 and 15-19 years. Diabetes was diagnosed using World Health Organisation criteria, based on an oral glucose tolerance test. The prevalence of diabetes increased over time in children aged 10 years and over: in boys from 0 % in 1967-1976 to 1.4% in 1987-1996 in the 10-14 year old age group, and from 2.43% to 3.78% for age group 15-19 and in girls from 0.72 % in 1967-1976 to 2.88 % in 1987-1996 in the 10-14 year old age group, and from 2.73 % to 5.31 % for age group 15-19 years. Along with the increase in the prevalence of Type II diabetes (p < 0.0001), there was an increase in weight (calculated as percentage of relative weight, p < 0.0001), and in frequency of exposure to diabetes in utero (p < 0.0001). The increasing weight and increasing frequency of exposure to diabetes in utero accounted for most of the increase in diabetes prevalence in Pima Indian children over the past 30 years. Type II diabetes is now a common disease in American Indian children aged 10 or more years and has increased dramatically over time, along with increasing weight. A vicious cycle related to an increase in the frequency of exposure to diabetes in utero appears to be an important feature of this epidemic. PMID- 9726593 TI - The incidence and characteristics of silent cerebral infarction in elderly diabetic patients: association with serum-soluble adhesion molecules. AB - The purpose of this study was to investigate the relationship between complications arising from silent cerebral infarction (SCI) and changes in the levels of serum-soluble adhesion molecules in 82 elderly diabetic patients aged 60 years and older. SCI was found in 43 % of the 82 patients, with incidence increasing in relation to age. The prevalence of SCI was higher in subjects with hypertension, poor metabolic control and increased fibrinolysis. The levels of soluble intercellular adhesion molecule-1 (sICAM-1), vascular cell adhesion molecule-1 (sVCAM-1) and E-selectin (sE-selectin) were higher in diabetic patients than in non-diabetic subjects (p < 0.05, p < 0.001, and p < 0.05, respectively). Also, sICAM-1 and sVCAM-1 were found at increased levels in diabetic patients with SCI compared to those without SCI (p < 0.01 and p < 0.05, respectively). In particular, the level of sICAM-1 was increased in patients with SCI due to perforating arterial occlusion, while the level of sVCAM-1 was increased in patients with SCI due to cortical arterial occlusion. However, no significant difference was found in sE-selectin levels. Overall average of the intima and media thickness (IMT) of the common carotid arteries increased with age. IMT proved to be greater in patients with SCI than in patients without SCI (p < 0.05), and showed a weak but significant positive correlation with sVCAM-1, while no correlation was found with either sICAM-1 or sE-selectin levels. In conclusion, measurement of serum adhesion molecules may be useful for diagnosing the early stages of brain damage and for prophylactic treatment which may prevent the onset or progression of SCI. PMID- 9726594 TI - Short-wavelength sensitive visual field loss in patients with clinically significant diabetic macular oedema. AB - The aim of the study was to compare the sensitivity of short-wavelength and conventional automated static threshold perimetry for the psychophysical detection of abnormality in patients with clinically significant diabetic macular oedema. The sample comprised 24 patients with clinically significant diabetic macular oedema (mean age 59.75 years, range 45-75 years). One eye of each patient was selected. Exclusion criteria included the presence of lenticular opacity. The sensitivity of the macular visual field of each patient was determined with programme 10-2 of the Humphrey Field Analyser on two occasions, using both short wavelength and conventional stimulus parameters; the results of the second session were analysed to minimise learning effects. A pointwise horizontal hemifield asymmetry analysis was derived for short-wavelength perimetry (thereby negating the influence of pre-receptoral absorption); the pointwise pattern deviation probability plot was analysed for conventional perimetry. Abnormality was defined as 3 or more contiguous stimulus locations with negative asymmetries (short-wavelength) or reduced sensitivity values (conventional) that resulted in a statistical probability level of p less than 0.05. The fields of 8 patients were abnormal as assessed by conventional perimetry while all were classified as abnormal using short-wavelength perimetry. In the 8 patients who exhibited both abnormal conventional and abnormal short-wavelength perimetry results, the extent of field loss was generally greater using short-wavelength perimetry. The position of the localised field loss (i.e. as distinct from field loss that was generalised across the visual field) assessed by short-wavelength perimetry corresponded with the clinical mapping of the area of diabetic macular oedema but the extent of this loss was generally greater than that suggested by clinical assessment. Short-wavelength automated perimetry offers improved sensitivity for the psychophysical detection of clinically significant diabetic macular oedema. PMID- 9726595 TI - Plasma phospholipid transfer protein activity is related to insulin resistance: impaired acute lowering by insulin in obese Type II diabetic patients. AB - Cholesteryl ester transfer protein (CETP) and phospholipid transfer protein (PLTP) have important functions in high density lipoprotein (HDL) metabolism. We determined the association of plasma CETP and PLTP activities (measured with exogenous substrate assays) with insulin resistance, plasma triglycerides (TG) and non-esterified fatty acids (NEFA), and assessed the lipid transfer protein response to insulin during a 6-7 h hyperinsulinaemic euglycaemic clamp in non obese and obese healthy subjects and patients with Type II (non-insulin dependent) diabetes mellitus (n = 8 per group). Plasma PLTP activity was higher in obese healthy subjects and obese Type II diabetic patients compared with non obese healthy subjects (p < 0.05 to 0.01) and was correlated with insulin resistance, plasma TG and NEFA (p = 0.02 to < 0.01). In non-obese healthy subjects, insulin decreased plasma TG and increased the HDL cholesteryl ester (CE)/TG ratio (p < 0.01 compared with saline infusion). Plasma PLTP activity fell by 14% at the end of the clamp (p < 0.01 compared with saline) but CETP activity did not change. The decreases in plasma NEFA, TG and PLTP activity and the rise in HDL CE/TG were smaller in obese Type II diabetic patients than in non-obese healthy subjects (p < 0.01 for all). Baseline HDL CE/TG was negatively correlated with plasma TG (p < 0.001, n = 32) and PLTP activity (p < 0.01) but not with CETP activity. Likewise, the rise in HDL CE/TG during the clamp was related to the fall in plasma TG (p < 0.001) and in PLTP activity (p < 0.02). It is concluded that plasma PLTP, but not CETP, is regulated by insulin in an acute setting. High plasma PLTP activity is associated with insulin resistance in conjunction with altered NEFA and triglyceride metabolism. High plasma TG and PLTP activity have coordinate effects on HDL metabolism. PMID- 9726596 TI - Reduced gene expression of UCP2 but not UCP3 in skeletal muscle of human obese subjects. AB - Massive overweight is an increasing health problem and underlies several complications which in turn result in premature death. The mechanisms underlying the imbalance between energy intake and energy expenditure, that lead to obesity in humans, are still only partly understood. In rodents, heat generation and the burning of calories by the mitochondrial uncoupling protein 1 (UCP1) are important for metabolic control. However, UCP1 is exclusively expressed in brown fat which is only present in limited amounts in human adults. The recent characterization of two new uncoupling proteins, UCP2 and UCP3, may elucidate potentially important pathways for energy expenditure regulation in man. The aim of this study was to investigate whether obesity is accompanied by aberrations in UCP2 and UCP3 regulation. Expression of these two genes was examined using in situ hybridization in six lean and six obese, but otherwise healthy, men. The UCP2 expression was decreased by 28 % (p = 0.001) in the abdominal muscle of the obese subjects. No differences in UCP3 expression were observed between obese and control subjects, although there was great variation in the expression between subjects. In conclusion, these data suggest an impaired activity of the mitochondrial uncoupling protein UCP2, but probably not UCP3, in obese subjects. This may result in decreased energy expenditure and contribute to the development and maintenance of obesity. PMID- 9726597 TI - Uncoupling protein-2 gene: reduced mRNA expression in intraperitoneal adipose tissue of obese humans. AB - The mitochondrial uncoupling protein-2 (UCP-2) is a recently discovered homologue of the brown adipose tissue-specific uncoupling protein and could be involved in the regulation of energy balance. Since obesity is associated with disturbed energy homeostasis, we tested the hypothesis that UCP-2 gene expression is deficient in this disorder. We determined, by a competitive reverse transcription polymerase chain reaction assay, UCP-2 mRNA expression in intra- and extraperitoneal adipose tissues of 107 morbidly obese subjects and 31 lean control subjects. In both obese and non-obese subjects, UCP-2 mRNA abundance was higher in the intraperitoneal than in the extraperitoneal tissue (p < 0.05), but no association was observed between intra- and extraperitoneal expression in either group. Compared with lean control subjects, both male and female obese subjects displayed significantly lower average UCP-2 mRNA expression in the intraperitoneal adipose tissue (p < 0.006), while UCP-2 mRNA abundance in extraperitoneal adipose tissue was not different between obese and non-obese men and women. Intraperitoneal UCP-2 mRNA remained low in nine obese subjects who lost 23 +/- 12 kg of weight over a period of 10 +/- 5 months subsequent to weight reducing surgery. These data support the concept that impaired adipose tissue expression of UCP2 may play a role in the pathophysiology of obesity. PMID- 9726599 TI - Spleen cells of non-obese diabetic mice fed with pig splenocytes display modified proliferation and reduced aggressiveness in vitro against pig islet cells. AB - A new means of modifying xenogeneic reaction to pig islet cells, which involves pre-feeding with pig spleen cells, was investigated for the first time in the non obese diabetic (NOD) mouse. Compared with controls, mice fed with pig spleen cells displayed much higher splenocyte proliferation in response to pig spleen and islet cells (p < 0.0001). This enhanced proliferation was specific for the species providing the fed cells. Positive relationships (p < 0.01) were found between increased splenocyte proliferation in response to pig spleen or islet cells and the number of cells per feeding or the number of daily feedings. Concomitantly, while co-incubation with splenocytes from control mice led to inhibition of both basal and stimulated insulin releases from pig islet cells (p < 0.001), this aggressiveness was abolished (p < 0.001) after co-culture with splenocytes from mice fed with pig spleen cells. The proliferative responses of splenocytes from fed or control mice to pig islet or spleen cells were abolished after removal of plastic-adherent cells, indicating that the major indirect pathway of T-cell activation was unchanged by pig spleen cell feeding. The main T splenocyte subsets involved were restricted to MHC class II as they did not proliferate in the presence of monoclonal antibodies (mAbs) directed at I-A molecules. In mice fed with pig spleen cells, as well as in control mice, the blocking of CD4 + T cells with mAbs led to abolition of proliferation (p < 0.002), while the blocking of CD8 + led to a less marked effect. However, an increase in the blocking effect of anti-CD8 mAbs was noted in mice fed with pig spleen cells (p < 0.02). In control mice, the main splenocyte subset involved during proliferation in response to pig islet cells was Thl, since interferon gamma (IFNgamma) production increased significantly (p < 0.01) while that of interleukin-10 (IL-10) increased only slightly. The main change observed in mice fed with pig spleen cells was a marked increase in basal IL-10 production (p < 0.01) and the basal IL-10/IFNgamma ratio (p < 0.001). It seems likely that feeding with pig spleen cells shifted the Th1/Th2 balance towards a dominance of Th2-type class II-restricted CD4 + T cells, which may have been conducive to activating CD8 + suppressor T cells. In any event, oral administration of pig cells modified xenogeneic cellular response, which may have implications for xenografts of pig islets. In a more general sense, physiological feeding of cells from xenogeneic species would appear to have certain effects on the immune system. PMID- 9726598 TI - Mannitol 1-phosphate mediates an inhibitory effect of mannitol on the activity and the translocation of glucokinase in isolated rat hepatocytes. AB - When tested in the presence of an inhibitor of sorbitol dehydrogenase, both mannitol and sorbitol caused a progressive inhibition of the detritiation of [2 3H]glucose in isolated rat hepatocytes. The purpose of the present work was to investigate the possibility that this effect was mediated by the regulatory protein of glucokinase. When added to hepatocytes, mannitol decreased the apparent affinity of glucokinase for glucose and increased the concentration of fructose required to stimulate detritiation, without affecting the concentration of fructose 1-phosphate. Its effect could be attributed to the formation of mannitol 1-phosphate, a potent agonist of the regulatory protein, which, similarly to fructose 6-phosphate, reinforces its inhibitory action. Formation of mannitol 1-phosphate in hepatocytes was dependent on the presence of mannitol and was stimulated by compounds that increase the concentration of glucose 6 phosphate. Liver extracts catalysed the conversion of mannitol to mannitol 1 phosphate about 7 times more rapidly in the presence of glucose 6-phosphate than of ATP. The glucose 6-phosphate-dependent formation was entirely accounted for by a microsomal enzyme, glucose-6-phosphatase and was not due to a loss of latency of this enzyme. In hepatocytes in primary culture, mannitol decreased the detritiation rate and counteracted the effect of fructose to stimulate glucokinase translocation. Taken together, these results strongly support a central role played by the regulatory protein in the control of glucokinase activity and translocation in the liver, as well as a feedback control exerted by fructose 6-phosphate on this enzyme. PMID- 9726601 TI - Common amino acid substitutions in insulin receptor substrate-4 are not associated with Type II diabetes mellitus or insulin resistance. AB - The family of insulin receptor substrates (IRS1-4) is defined by proteins with an overall similar structure. IRS-1 and IRS-2 have been shown to have key roles in cellular transmission of the action of insulin, insulin-like growth factor-1 and various cytokines. We have previously identified amino acid polymorphisms in the human IRS-1 and IRS-2 proteins. Given the documented importance of IRS-1 and -2 in insulin signalling and the implications of distribution of these genes for the pathogenesis of insulin resistance and diabetes, we decided that the most recently identified member of the IRS family, IRS-4, was a relevant candidate to examine for genetic variability which might be associated with subsets of diabetes or insulin resistance. The gene encoding IRS-4 was analysed by the single strand conformation polymorphism technique in 83 Danish Caucasians with Type II (non-insulin-dependent) diabetes mellitus. Five amino acid polymorphisms were identified: Leu34Phe, Arg411Gly, Gly584Cys, His879Asp and Lys883Thr. In an association study of 324 patients with Type II diabetes and 267 control subjects with normal glucose tolerance the polymorphism at codon 34 was found with allelic frequencies of 3.9 and 2.3 %, respectively, the variant at codon 411 with allelic frequencies of 3.9 and 5.6%, respectively, and the variant at codon 879 with frequencies of 19.2 and 18.0%, respectively. Each carrier of the codon 34 polymorphism was also a carrier of the codon 411 and codon 879 variants and similarly, carriers of the variant at codon 411 were also carriers of the polymorphism at codon 879. The variants at codon 584 and 883 were each found in only one Type II diabetic patient. The allelic frequencies of the variants at codon 411 and 879 were also determined in 380 young healthy subjects (4.6 and 18.1 %, respectively). The insulin sensitivity index as estimated by Bergman's minimal model of the young healthy subjects carrying either polymorphism was indistinguishable from the carriers of wild-type IRS-4. Moreover, none of the men were heterozygous for the IRS-4 polymorphisms indicating that the gene is located on the X-chromosome. In conclusion, amino acid polymorphisms in human IRS-4 are common in Caucasians but are not associated with Type II diabetes or with insulin resistance in young healthy subjects. PMID- 9726600 TI - Pioglitazone-induced increase of insulin sensitivity in the muscles of the obese Zucker fa/fa rat cannot be explained by local adipocyte differentiation. AB - Thiazolidinediones are potent antidiabetic compounds, which act by enhancing peripheral insulin sensitivity. They are also activators of the peroxisome proliferator activated receptor gamma in adipose tissue. Pioglitazone induces in vivo adipocyte differentiation in the obese Zucker fa/fa rat and hence the capacity of adipose tissue to utilize glucose. Nevertheless, muscles are the major site for insulin-mediated glucose disposal. The increase of muscle glucose utilization under thiazolidinedione treatment could be secondary to local adipose tissue differentiation. This possibility is supported by the fact that a thiazolidinedione-induced myoblast conversion into adipocytes has been described in vitro. To address this problem, we have studied the in vivo effect of a pioglitazone treatment on insulin-induced glucose utilization and the expression of genes exclusively expressed in mature adipocytes in three muscles differing by their fibre composition in Zucker (fa/fa) rats. Whereas pioglitazone treatment increased insulin-stimulated glucose utilization to the same extent in all muscle types, an adipocyte differentiation was only present in the oxidative muscle, the soleus. Soleus muscle was also the only one in which the presence of genes specific for adipose tissue could be detected before the pioglitazone treatment. There was no detectable expression of adipocyte specific genes in the extensor digitorum longus or in the epitrochlearis muscles before or after the drug treatment. We conclude that pioglitazone effects on muscle glucose metabolism cannot be due to a local adipocyte differentiation, and that the conversion of myoblasts into adipocytes under thiazolidinedione stimulation observed in vitro is, if it exists, a marginal phenomenon in vivo. PMID- 9726602 TI - Glycosaminoglycan therapy for long-term diabetic complications? PMID- 9726603 TI - Fosinopril decreases levels of soluble vascular cell adhesion molecule-1 in Type II diabetic patients suffering from microalbuminuria. PMID- 9726604 TI - Dramatic recovery of counter-regulatory hormone response to hypoglycaemia after intensive insulin therapy in poorly controlled Type I diabetes mellitus. PMID- 9726605 TI - Hereditary haemochromatosis mutations (HFE) in patients with Type II diabetes mellitus. PMID- 9726606 TI - Serum levels of vascular endothelial growth factor in diabetic subjects: the relationship with blood pressure. PMID- 9726607 TI - Clinical experience with heart transplantation in infants. AB - OBJECTIVE: Orthotopic heart transplantation has become an accepted therapeutic concept for adult patients with endstage heart disease. In newborns and infants this procedure is still a matter of discussion because of unknown long-term results and the lack of donor organs. METHODS: Since March 1988 we have performed 40 orthotopic heart transplantation in 39 infants who were from 1 to 280 days of age. Indications for transplantation included hypoplastic left-heart syndrome (n = 28), dilative cardiomyopathy (n = 4), endocardial fibroelastosis (n = 4) and other complex structural anomalies (n = 3). The mean waiting period for transplantation was 53 days. A donor-recipient weight ratio up to 4.0 was accepted. Profound hypothermic circulatory arrest was used for graft implantation in all those patients who required extensive aortic arch reconstruction (71%). The initial immunomodulation was based on Cyclosporine, Azathioprine and Prednisolone. Patients who underwent transplantation during the first 6 weeks of life received a chronic single-drug therapy with Cyclosporine after 1 year. RESULTS: There were six peri-operative deaths caused by drug-resistant right heart failure in three cases, humoral rejection (n = 1), CMV infection (n = 1) and multi organ failure (n = 1). One infant died late, due to rejection. The actuarial survival rate for the entire group is now 82%. There is a remarkable influence of increasing experience. Whereas six of 15 infants who had heart transplantation between 1988 and 1993 died early post-operatively (survival rate: 60%), only one late death occurred among 24 recipients in the period from 1994 to April 1997 (survival rate: 96%). Episodes of rejection occurred once or several times in about half of the patients in this series (48%). All surviving children are living at home in excellent condition. CONCLUSIONS: Heart transplantation during early infancy is a rational and durable therapy for heart diseases with irreversible myocardial failure or severe structural anomalies. The intermediate term results have been encouraging in many centers, but more data must be accumulated to determine the sequelae of chronic immunosuppression. The lack of donor organs remains one of the major problems in pediatric heart transplantation. PMID- 9726608 TI - Orthotopic cardiac transplantation for the failing Fontan circulation. AB - OBJECTIVE: Modified Fontan procedures are now employed in several conditions unsuitable for bi-ventricular repair. Selection criteria have been relaxed. The procedure is palliative. Longterm outlook is unknown. This study evaluated factors associated with the development of a failing Fontan circulation and transplantation results. METHODS: Retrospective review of patients referred to a single centre for cardiac transplant assessment. RESULTS: Between 1985 and 1996, 46 of 448 cardiac transplants were performed for congenital heart disease. Nine of these were performed in patients with a failing Fontan circulation (four adults, five children). In six cases, the dominant ventricle had left ventricular (LV) morphology. Congenital anomalies included double outlet right ventricle (three cases), double inlet left ventricle (two cases), tricuspid atresia (two cases), and pulmonary atresia with intact ventricular septum (one case). Fontan procedures were performed in absence of sinus rhythm (four cases), atrio ventricular (AV) valve regurgitation (two cases), aortic regurgitation and systolic LV dysfunction (one case), elevated mean pulmonary artery pressure (one case), and older age (>7 years, eight cases). Three patients required early re operation and two needed permanent pacing. Subsequent deterioration associated with loss of sinus rhythm (four cases) and progressive AV valve regurgitation (seven cases) led to transplant assessment (at < 1 year, five cases; at 2-12 years, four cases). All patients were listed for transplantation. Three patients required intravenous inotropic support and three patients with lymphocytotoxic antibodies needed prospective crossmatching. Donor cardiectomy was modified to facilitate implantation. The recipient operation involved pulmonary artery reconstruction (using pericardium), modified atrial and direct caval anastomoses. Three patients died within 24 h of surgery (two graft failures, one haemorrhage). In operative survivors (n = 6), intensive care stay was 3-16 days, and hospital stay ranged from 14 to 32 days. There have been no subsequent deaths (follow up, 0.5-4.7 years). CONCLUSION: In high-risk Fontan candidates, transplantation may be preferable at the outset. Previous surgery, lymphocytotoxic antibodies, indeterminate pulmonary vascular resistance, emergency status, sub-optimal donor selection, and perioperative bleeding contribute to peri-operative mortality. In survivors, the outcome remains very encouraging. PMID- 9726609 TI - Is routine post-operative surveillance for cytomegalovirus infection following heart transplantation necessary? AB - OBJECTIVE: Cytomegalovirus infection (CMV) is an important cause of morbidity and mortality following cardiac transplantation. The purpose of the present study was to ascertain whether routine post-operative screening for CMV infection influenced clinical management. METHODS: Laboratory and case notes of 220 patients who received cardiac transplantation between November 1986 and October 1996 were reviewed. The range of follow-up was one to 120 (median 36) months. CMV surveillance involved blood tests for early antigen detection weekly for the first 6 post-operative weeks, fortnightly thereafter until the end of the third post-operative month and every 6 weeks to the end of the first post-operative year. Otherwise monitoring was performed if the patients had clinical symptoms suggestive of CMV infection. CMV sero-negative IgG recipients (R) of sero positive IgG donor (D) organs and/or blood products received hyper-immune gammaglobulin for the first three post-operative months. Four patient groups were noted, namely R+D+ (59 patients), R+D- (70 patients), R-D+ (35 patients) and R-D- (56 patients). RESULTS: CMV antigenaemia was present in 40% (89) of patients and 48% (43) of these patients developed clinical features of CMV infection and received ganciclovir therapy. The distribution of clinical CMV infection requiring treatment was 25% (9/35) in the R+D- group, 50% (16/32) in the R+D+ group and 85% (18/22) in the R-D+ group. None of the patients in the R-D- group developed CMV antigenaemia. Forty six (52%) patients who were CMV antigen positive but who did not develop symptoms were not treated with ganciclovir and have remained well. CONCLUSION: Our results suggest that routine screening for CMV following cardiac transplantation is unnecessary. Surveillance did not result in the instigation of treatment for CMV unless there were associated clinical features of CMV infection. PMID- 9726610 TI - Surgery for coarctation of the aorta in infants younger than 3 months: end-to-end repair versus subclavian flap angioplasty: is either operation better? AB - OBJECTIVE: Recurrent coarctation is a complication which is seen at a consistent rate following all types of repair for coarctation of the aorta. Particularly disappointing late results are reported in younger infants, under 3 months of age. This retrospective analysis was undertaken to compare the outcomes on late follow-up between subclavian flap angioplasty and resection and end-to-end repair, in this age group. METHODS: Over a 12-year period, between 1982 and 1994, 86 infants under 3 months of age underwent surgical repair of coarctation (39 resections and end-to-end repair, and 47 subclavian flap angioplasty procedures). Operative mortality was not significantly different (P = 0.6) between resection and end-to-end repair (5.1%) and subclavian flap angioplasty (8.5%). All operative deaths (six patients) were in infants with associated ventricular septal defects. The mean follow-up for all patients was 7.95 years +/- 4.10 (range 0-14.5 years). The 5-year survival for resection and end-to-end repair was 87 +/- 5%, compared to 75 +/- 7% for subclavian flap angioplasty (P = 0.2). RESULTS: Recurrent coarctation occurred in nine patients who needed reoperation. The reoperation-free rates at both 5 and 10 years for resection and end-to-end anastomosis, and subclavian flap repair were 86 +/- 6% and 90 +/- 5%, respectively. The recurrence in the resection and end-to-end anastomosis group were due to constrictive scarring at the anastomosis, whereas periductal tissue and growth of posterior aortic ridge caused recurrence in the subclavian flap angioplasty group. There were no deaths during reoperation for recurrence. CONCLUSIONS: Both procedures are extremely effective for coarctation repair in young infants and run a similar risk of recurrence, which are due to completely different mechanisms. The surgeon's expertise is the major determinant of outcome. PMID- 9726611 TI - Lung volume reduction or lung transplantation for end-stage pulmonary emphysema? AB - OBJECTIVE: As the waiting period for lung transplant (LT) candidates with end stage pulmonary emphysema (COPD) continues to increase, there is a need for alternative treatments to reduce the morbidity and mortality associated with COPD. We hypothesized that lung reduction (LR) may avoid the need for subsequent LT in patients on the waiting list that are also candidates for LR. METHODS: From July 1994 to December 1995, 20 patients received LR as alternative to LT. The average age was 58 +/- 7 years; 11 were males. Eighteen patients had primary COPD and two had alpha-1 antitrypsin deficiency. Eighteen LRs were thoracoscopic (two bilateral and 16 unilateral) and two were done through a median sternotomy. RESULTS: At a follow-up of 32 +/- 4 months, 19 patients are alive (19/20 = 95%). Fifteen patients (15/20 = 75%) are currently off the LT list and doing well: FEV1 is 40 +/- 18% predicted at 2 years compared with 22.7 +/- 6% before LR (P < 0.001); FVC is 84 +/- 13% at 2 years compared with 55 +/- 7% (P < 0.001) and the RV is 145 +/- 59% compared with 270 +/- 58% (P < 0.001). One patient (5%) required extra-corporeal membrane oxygenation (ECMO) after LR to the contralateral side of the first procedure and subsequently died. Two patients (10%) are currently listed for LT because of persistent symptoms. One patient (5%) in whom deterioration was secondary to exposure to toxic fumes, underwent successful LT. One patient (5%) is doing well from the pulmonary standpoint but is being worked up for new severe coronary artery disease (CAD). The freedom from LT is 95% (19/20) and the freedom from repeat LR is 85% (17/20). CONCLUSIONS: LR has the potential to offer an effective palliative alternative to LT in 75% of selected patients up to 32 months of follow-up. Widespread use of bilateral LR is anticipated to further improve the results. PMID- 9726612 TI - Unilateral thoracoscopic reduction pneumoplasty for asymmetric emphysema. AB - OBJECTIVE: We prospectively analyzed the surgical and functional results of unilateral thoracoscopic reduction pneumoplasty which we performed by choice in patients with asymmetric emphysema. METHODS: Between October 1995 and June 1997, 119 emphysematous patients were examined and 34 were operated upon. Among these, 14 selected patients with asymmetric distribution of emphysema in the lungs underwent unilateral reduction pneumoplasty (ten right, and four left). There were 13 males and one female, with a mean age of 62 years. Eligibility criteria included bullous and non-bullous end-stage emphysema with severe limitation to daily activity. RESULTS: No patient required conversion to thoracotomy. Mean operative time ranged between 70 and 240 min with a mean of 103 min. There was no postoperative mortality but five patients developed one or more complications: five prolonged air leaks (>7 days); two pulmonary infections; one empyema. No patient required postoperative mechanical ventilation. Median hospital stay was 8 days. At the 3-month follow-up the mean FEV1 increased from 0.8 l to 1.2 l (P < 0.001). Mean FVC increased from 2.6 l to 2.9 l (P < 0.001). The Medical Research Council dyspnea score decreased from a mean of 3.2 to 1.8 (P < 0.001). CONCLUSIONS: Asymmetric distribution is a frequent finding in patients with severe emphysema. Unilateral thoracoscopic reduction pneumoplasty may represent an ideal approach in this selected group of patients. PMID- 9726613 TI - Pulmonary resection for malignancy in the elderly: is age still a risk factor? AB - OBJECTIVE: There is an increasing number of elderly patients presenting with potentially-resectable lung malignancy. The objective of this study is to evaluate the modern perioperative morbidity and mortality in patients undergoing oncologic lung resection and to analyse the trend over a 26-year period in our experience. METHODS: Between 1971 and 1996, 1506 patients underwent lung resection for malignancy. We reviewed the 30-day perioperative risk in a group of 385 (25.6%) patients aged 70 years and older operated on for intended cure of lung malignancy. Operations included 293 (77%) lobectomies, 24 pneumonectomies (6%), 16 bilobectomies (4%) and 52 wedge or segmental resections (13%). The pathology was bronchogenic carcinoma in 89% and metastasis or other tumours in 11% of patients. We compared the 30-day perioperative risk between the elderly group (age 70 or greater) and a cohort of 180 patients (control) 69 years and younger. RESULTS: The mortality for all resections in elderly group was 4.2% (16/385) and was 1.6% for the control group. Mortality in the octogenarian group was 2.8%. Female gender correlated with a decreased risk of death, with only two of 16 deaths in females (P < 0.005). Overall morbidity was higher in the study than in control patients (34% vs. 25%, n.s.), although major morbidity was similar in both groups (13.2% vs. 13%). Abnormal pulmonary-function testing and positive cardiac history did not correlate with increase overall or specific risk. Pneumonectomy carried a higher risk for death, with three of 24 deceased (12.5%; P < 0.05). Changes in outcome were analysed over two time periods: the mortality in the early period (1971-1982), 11.1% (8/72), was significantly elevated above the control group, while mortality in the modern period (1983 1994) was not, with a rate of 2.6% (8/313). CONCLUSIONS: In our series, mortality associated with operative treatment for lung malignancy in the elderly declined, so age alone no longer appears to be a risk factor. Age remains a risk factor for overall, but not major, morbidity. Pneumonectomy should undertaken cautiously in this age group. Based on this data, functional elderly patients should not be denied curative lung resection based on age alone. PMID- 9726614 TI - Postoperative complications after bronchoplastic procedures in the treatment of bronchial malignancies. AB - OBJECTIVE: The purpose of this study was to determine the frequency of postoperative complications after bronchoplastic procedures in the treatment of pulmonary malignant tumors and to analyze the factors influencing the complication rate. METHODS: During a 5-year-period (1992-1996) 79 patients (68 male, 11 female, mean age 57 years) underwent reconstructive operations for bronchial malignancies. We performed 58 bronchoplastic procedures and 21 combined broncho- and angioplastic procedures. Among the bronchoplastic procedures the number of sleeve resections (n = 44) and wedge resections (n = 35) were comparable. RESULTS: Fifty-nine patients (74,7%) showed a regular postoperative course; 12 patients (15,2%) had severe postoperative complications (death, re operation). Concerning the primary operation the operative 30-day mortality was 5.1% (n = 4) and including the two deaths after re-operation it was 7.6% (n = 6). After subdividing the patients into three groups (severe, less severe and no complications) we tried to determine predictors for occurrence of postoperative complications. There was a higher rate of severe complications in the age group 61-70 years (6/25 = 24%) as compared with younger patients between 51 and 60 years (4/38 = 10,5%; P < 0,05). Concerning the location, the outcome was better after sleeve- or wedge lobectomies of the upper lobes (four complications/51 patients = 7.8%) compared with procedures of the lower lobes (3/14 = 21.4%). The data could not prove a lower frequency of severe postoperative complications or specific morbidity after pleural coverage following bronchial sleeve resection. The complication rate was higher when sleeve resection of the bronchus was performed (10/44 = 22.7%) as compared with wedge resections (2/35 = 5.7%; P = 0,011) and after resection of T3/T4 tumors (6/28 = 21,4%) compared with T1/T2 tumors (4/37 = 10.8%; P < 0,05). CONCLUSIONS: Bronchoplastic procedures represent a fairly safe therapy opportunity in patients with centrally localized bronchial carcinoma and compromised or uncompromised pulmonary function. In this study the complication rate was higher after sleeve resection of the bronchus as compared with wedge resection. Pleural coverage of the anastomosis was not effective to prevent major complications due to dehiscence of the bronchial anastomosis. A pedicled muscle flap could be a valuable alternative. PMID- 9726615 TI - Ventricular remodelling and revascularization in severe left ventricular dysfunction. AB - OBJECTIVE: To evaluate the role of surgical revascularization in the presence of severe, global impairment of left ventricular function without discrete aneurysm formation or mitral regurgitation. The high mortality and morbidity associated with this group, together with the limited benefits tend to prompt referral for cardiac transplantation. METHODS: Fifty-three patients initially referred for transplantation, in addition to coronary revascularization, underwent mitral annuloplasty (group A = 23), free wall remodelling by endoaneurysmorrhaphy (group B = 17) or mitral annuloplasty and free wall reconstruction (group C = 13). The mean ages were 59, 56 and 57 years for groups A, B and C, respectively. Detailed assessment of pre- and post-operative physical and psychological status were carried out. RESULTS: Follow-up was for a mean period of 22-26 months. All patients reported substantial improvement in quality of life, both physical and psychological parameters and in NYHA functional class status. Objective evidence of improvement in ejection fraction was seen in all three groups but especially in group A. There were five early deaths, four were due to inadequate revascularization due to the poor quality of target vessels. There were three late deaths and one patient that required transplantation. CONCLUSION: We conclude that patients with severe left ventricular dysfunction can be candidates for surgical revascularization and optimization of ventricular geometry with acceptable mortality. The importance of achieving complete revascularization is emphasized in this series. PMID- 9726616 TI - Elimination of cardiopulmonary bypass: a prime goal in reoperative coronary artery bypass surgery. AB - OBJECTIVE: The purpose of this study was to evaluate morbidity and mortality in reoperative coronary artery bypass surgery using the New York State database. METHODS: Patients undergoing reoperative coronary artery bypass between January 1995 and December 1996 were included. Patients were operated using cardiopulmonary bypass (CPB group, n = 184) or without cardiopulmonary bypass (non-CPB group, n = 105) by surgeon preference. Groups were compared for preoperative risk factors, postoperative mortality and major complications. RESULTS: Crude mortality was lower in the non-CPB group, despite a higher expected mortality, resulting in a risk-adjusted mortality of 1.3% versus 2.7% for the CPB group (NS). Of non-CPB patients, 91.4% were without complications, while only 72.1% of CPB patients (P < 0.0001) were complication-free. Major complications were significantly reduced in non-CPB patients compared to CPB patients: stroke 0% versus 3.8% (P < 0.04), cardiovascular complications 4.8% versus 15.8% (P < 0.005), other major complications 1.9% versus 10.4% (P < 0.007). Postoperative IABP support was needed in 1.9% of the non-CPB group patients and in 14.2% of the CPB group (P < 0.0007). CONCLUSIONS: The main object of reoperative CABG is to relieve symptoms, since the survival benefit of the procedure has not been demonstrated. Performance of reoperative coronary artery bypass surgery without cardiopulmonary bypass significantly reduces morbidity. We conclude that cardiopulmonary bypass should be avoided whenever possible in reoperative coronary bypass surgery. PMID- 9726617 TI - Non-invasive measurement of cardiac output during coronary artery bypass grafting. AB - OBJECTIVE: A new device, using whole body bioresistance measurements and a new equation for calculating stroke volume has been developed. Using this equation, an attempt was made to correlate whole body bioresistance cardiac output with thermodilution cardiac output in patients undergoing coronary artery bypass grafting. METHODS: Thirty-one adults undergoing elective coronary artery bypass grafting were studied prospectively. Simultaneous paired cardiac output measurements by whole body bioresistance and thermodilution were made at five time points during coronary artery bypass grafting: in anesthetized patients before incision (T1), after sternotomy (T2), after opening the pericardium (T3), ten min post bypass (T4), and in the intensive care unit (T5). The patients had a mean of three thermodilution cardiac outputs compared with a mean of three bioimpedance measurements at each time point. The bias and precision between the methods were calculated. RESULTS: There was good correlation between bioresistance cardiac output (nCO) and thermodilution cardiac output (ThCO) measurements in both groups for all recorded times. The patients' mean ThCO and nCO, as well as bias and precision between methods were calculated. Mean ThCO ranged between 4.14 and 5.06 l/min; mean nCO ranged between 4.12 and 4.97 l/ min. Bias calculations ranged between -0.072 and 0.104 l/min. Precision (2 SD) calculations ranged between 0.873 and 1.228 l/min for 95% confidence intervals. Pearson's correlation ranged from 0.919 to 0.938. CONCLUSIONS: Cardiac output measured with the new device correlates well with the thermodilution measurements of cardiac output during and immediately following coronary artery bypass grafting. The overall agreement between the two methods was good. The new device is an accurate non-invasive method of measuring cardiac output during coronary artery bypass grafting. PMID- 9726618 TI - Calcium sensitization as new principle of inotropic therapy in end-stage heart failure? AB - OBJECTIVE: Due to shortage of donor hearts and increasing waiting-lists of patients with end-stage heart disease, new pharmacological principles for bridging therapies are necessary. The positive inotropic effects of cAMP increasing drugs (e.g. catecholamines, phosphodiesterase-inhibitors) are diminished in the failing myocardium. Hence, we investigated the usefulness and mechanism of the two calcium sensitizers, levosimendan and CGP 48506 in preparations from end-stage failing human hearts since the exact mechanism of the positive inotropic effects is not yet clearly understood. METHODS: Failing human hearts which required orthotopic heart transplantation due to idiopathic dilated cardiomyopathy were investigated. Contraction experiments were performed using muscle strips of ventricles. Calcium sensitization was investigated in skinned fibers and phosphodiesterase activity was measured in ventricular homogenate. In addition, cAMP levels were quantified in myocytes from guinea-pig hearts. RESULTS: In muscle stripes from failing human hearts levosimendan (10 micromol/l) increased the force of contraction only to 112.8 +/- 6.7% of predrug values. In contrast, CGP 48506 increased the force of contraction to 311 +/- 59% of predrug values at 100 micromol/l. The time to peak tension and time of relaxation were increased to 175 +/- 4% and 205 +/- 15% of control levels at 100 micromol/l. Skinned fibers from failing human hearts were sensitized to calcium with an EC50 of 10 micromol/l. Other mechanisms of action were excluded since CGP 48506 affected neither the activity of phosphodiesterase isoenzymes I-IV in failing human hearts, nor cAMP levels in guinea-pig cardiomyocytes. On the other hand, levosimendan (1 micromol/l) increased cAMP content from 6.3 +/- 0.3 to 8.1 +/- 0.3 pmol/mg protein. CONCLUSION: CGP 48506 is an inotropic agent with calcium sensitizing properties in the human heart, that is devoid of inhibitory activity on human cardiac phosphodiesterase isoenzymes. It offers, therefore, a new form of positive inotropic therapy that can be useful for the bridging treatment of heart failure before transplantation. On the other hand, levosimendan is a calcium sensitizer showing less-effective inotropic effects accompanied by increased cAMP levels. PMID- 9726619 TI - Effects of adenosine infusion with or without leukocyte depletion on recovery after hypothermic ischemia in neonatal lamb hearts. AB - OBJECTIVE: Leukocytes have been shown to have an important role in ischemia/reperfusion injury. Adenosine also reduced this ischemia/reperfusion injury. There is an interaction between adenosine and leukocyte via receptor mediated function. To determine whether beneficial effects of adenosine on reperfusion injury is mediated by changes in leukocyte function, we studied the effects of adenosine with and without leukocyte depletion during reperfusion on the functional recovery of the neonatal myocardium after cold cardioplegic arrest. MATERIALS AND METHODS: We infused adenosine (350 micromol/l) during the first 20 min of reperfusion for adenosine treated group and adenosine-leukocyte treated group. The other two groups were perfused with leukocyte treated blood or untreated blood. All the groups were subjected to 2 h of cold cardioplegic ischemia (n = 8 in each group). At 30 min of reperfusion, LV function was measured. Coronary blood flow and oxygen consumption (MVO2) were also measured to evaluate the metabolic recovery. RESULTS: Adenosine treated, adenosine-leukocyte treated, and leukocyte treated groups showed better functional recovery than the control group (maximum developed pressure: control = 74.6 +/- 5.6%, adenosine treated = 97.6 +/- 9.5%, adenosine-leukocyte treated = 98.5 +/- 5.6%, leukocyte treated = 82.5 +/- 6.0%. P < 0.05). Both adenosine treated and adenosine leukocyte treated groups showed better recovery than leukocyte treated group (P < 0.05). Coronary blood flow was higher in adenosine-leukocyte treated group compared to other groups (P < 0.05). MVO2/beat was higher in adenosine treated, adenosine-leukocyte treated, and leukocyte treated groups than control group (P < 0.05). CONCLUSION: Adenosine, with or without leukocyte depletion, had similar beneficial effect on recovery of systolic and diastolic functions, which involved other mechanisms in addition to the leukocyte inhibitory effect. PMID- 9726620 TI - Neuroprotective effects of preischemia subcutaneous magnesium sulfate in transient cerebral ischemia. AB - OBJECTIVE: Neurological injury due to transient cerebral ischemia is a potential complication of cardiovascular surgery. The neuroprotective effect of magnesium, when given subcutaneously before the ischemia, was assessed in a rat model of transient global cerebral ischemia. METHODS: Thirty-six male Wistar albino rats were included to this randomized, controlled, prospective study. In 24 animals, ischemia was induced with four-vessel occlusion technique with the duration of 15 min. MgSO4 was given 600 mg/kg subcutaneously 48 h before the procedure in group 1 (n = 12). Similar volume of saline solution was used in animals of control group (group 2, n = 12). The animals in group 3 (sham group, n = 12) were anesthetized and subjected to operative dissections without vascular occlusion. Physiological parameters and somatosensory evoked-potentials (SEP) were monitored in animals before ischemia, during ischemia and in the first 30 min of reperfusion. Their neurological outcome had been clinically evaluated and scored up to 4 days postischemia. The intergroup differences were compared. Then the animals were sacrificed and their brains were processed for histopathological examination. RESULTS: In group 3, SEP amplitudes did not change during the procedures, and all animals recovered without neurologic deficits. At the end of ischemic period, the average amplitude was reduced to 5 +/- 3% of the baseline in all ischemic animals. This was followed by a gradual return to 87 +/- 10% and 83 +/- 8% of the initial amplitude after 30 min of reperfusion in group 1 and group 2, respectively (P > 0.05). The average neurological score was significantly higher in group 1 than in group 2 at 48, 72 and 96 h after the ischemic insult (P < 0.05). Histological observations were clearly correlated with the neurological findings. CONCLUSION: The results suggest that subcutaneous MgSO4 reduces cerebral injury and preserves neurologic function when given two days before the transient global ischemia in rats. PMID- 9726621 TI - Medial smooth muscle cell loss in arterial allografts occurs by cytolytic cell induced apoptosis. AB - OBJECTIVE: Experimental arterial allografts, used as models of chronic rejection, undergo marked loss of smooth muscle cells (SMC) from their media prior to the development of occlusive, intimal proliferative lesions. Medial SMC loss has been described in human heart transplants, and may be related to the development of occlusive coronary lesions which are the hallmark of chronic rejection. This SMC loss does not exhibit the characteristics of necrotic cell death. We sought to determine whether medial SMC loss in arterial allografts occurs by apoptosis. We further investigated these allografts for cytolytic cell-derived inducers of apoptosis, Finally, we compared two different strain combinations to assess the impact of varying histoincompatability on medial SMC loss. METHODS: Evidence for internucleosomal DNA degradation, which is characteristic of apoptosis, was sought by the in situ terminal deoxynucleotidyl transferase nick end labelling (TUNEL) method carried out on Lewis to Fisher rat femoral artery transplants (disparate at minor loci only) and Brown Norway to Lewis aortic transplants (fully disparate at major and minor loci). Isografts (Lewis to Lewis) served as controls. In a separate series of experiments graft mRNA was extracted and analysed by reverse transcription-polymerase chain reaction (RT-PCR) with primers for molecular inducers of apoptosis (TNF-alpha, Fas ligand, perforin, and granzyme-B) which are derived from cytolytic cells known to be present in allografts. RESULTS: Allograft media contained large numbers of TUNEL stained nuclei in both strain combinations. Neither isografts nor ungrafted femoral artery segments stained positive for apoptosis. RT-PCR on whole allografts in both strain combinations revealed sustained upregulation of perforin, granzyme-B, Fas-ligand and TNF-alpha mRNA concomitant with medial SMC loss. Autografts demonstrated sustained up regulation of TNF-alpha, and perforin, but only brief upregulation of granzyme-B, and no upregulation of Fas-ligand. CONCLUSIONS: These data strongly suggest that medial SMC loss in allograft arteriopathy occurs by apoptosis. Further, RT-PCR data indicate that cytolytic cell-derived inducers of apoptosis are upregulated in these grafts and may be accountable for medial SMC apoptotic cell death. Finally, fully-disparate (Brown Norway to Lewis) and minor only incompatible (Lewis to Fisher) strain combinations both resulted in marked intimal proliferation, medial SMC loss by apoptosis, and similar patterns of expression of cytolytic cell derived inducers of apoptosis. Insofar as intimal proliferative lesion-formation may be dependent on medial damage (as in arterial injury models), understanding the mechanism of medial SMC loss may provide a novel therapeutic approach to human cardiac transplant arteriopathy. PMID- 9726622 TI - Extrapericardial solitary fibrous tumour of the pericardium. AB - Solitary fibrous tumour (SFT) occurs most commonly in the pleura and is extremely rare in the pericardium. The authors report a case of a 60-year-old man in whom a large mediastinal mass was accidentally discovered. Computed tomography showed involvement of the left anterosuperior mediastinum with displacement of the trachea, large vessels and oesophagus; histopathological findings after complete resection of the neoplasia demonstrated an SFT of the pericardium, the first reported case with extrapericardial pattern of growth. A review of the literature on SFTs of the pericardium is provided. PMID- 9726623 TI - Use of the Harmonic Scalpel for harvesting arterial conduits in coronary artery bypass. AB - A simple and effective technique is described here for harvesting the gastroepiploic artery (GEA) and radial artery (RA) using the Harmonic Scalpel. The mean time of harvesting GEA was 9 min and that of RA was 17 min. There were no injuries or spasms of those grafts and the postoperative angiograms performed in 28 patients. This shows 100% patency of the conduits. The GEA and RA are safely harvested by using the Harmonic Scalpel and the use of arterial conduits in coronary artery bypass grafting (CABG) seems to be easily achieved. PMID- 9726624 TI - Involvement of delta-opioid receptors in the effects induced by endogenous enkephalins on learned helplessness model. AB - Pharmacological, neurochemical and behavioural findings support a possible role of endogenous opioids in clinical depression. There is evidence from animal studies that delta-opioid receptors are involved in several behavioural responses to opioids, including motivational activities. In the present study, the mixed enkephalin catabolism inhibitor, RB 101 (N(R,S)-2-benzyl-3[(S)-(2-amino-4 methylthiobutyldithio]-1-oxoprop yl)-L-phenylalanine benzyl ester) (1.25, 2.5 and 5 mg/kg), induced a dose-dependent antidepressant-like effect in a learned helplessness model. Thus, RB 101 reversed escape deficits in rats previously subjected to inescapable shocks, suggesting the involvement of endogenous enkephalins in depression. Similar effects were observed after administration of the selective delta-opioid receptor agonist, BUBU (Tyr-D.Ser-(O-tert-butyl)-Gly Phe-Leu-Thr(O-Tet-butyl-OH) (1 and 2 mg/kg). Moreover, RB 101 effects were antagonized by administration of naltrindole (NTI) (0.1 mg/kg), which points to a preferential involvement of delta-opioid receptors in this enkephalin-controlled behaviour. As RB 101 has been reported to be almost devoid of opiate-related side effects, it could represent a promising alternative in the treatment of depressive patients who are unresponsive to, or intolerant of, classical antidepressants. PMID- 9726625 TI - Effects of pre-exposure and co-administration of the cannabinoid receptor agonist CP 55,940 on behavioral sensitization to cocaine. AB - Rats given cocaine (15 mg/kg, i.p.) every second day over a 2-week period displayed a progressively greater locomotor response to the drug over days indicating behavioral sensitization. When the cannabinoid receptor agonist CP 55,940 ((-)-cis-3-[2-hydroxy-4-(1,1-dimethylheptyl)phenyl]-trans-4-(3-hyd roxypropyl)cyclohexanol) (10, 25 or 50 microg/kg) was administered under a similar regime, no such sensitization was observed. Rather, the two highest doses of CP 55,940 (25 and 50 microg/kg) caused locomotor suppression that lasted throughout administration. When rats pre-exposed 10 times to CP 55,940 were challenged with cocaine (15 mg/kg), no exaggerated locomotor response to cocaine was evident relative to non pre-exposed rats. When these rats were subsequently re-tested with CP 55,940, the cannabinoid continued to produce a dose-dependent suppression of locomotor activity. Finally, when CP 55,940 (50 microg/kg) was co administered with cocaine, it significantly reduced the locomotor hyperactivity produced by the drug but did not block the development of behavioral sensitization. These results show that CP 55,940 does not sensitize locomotor activity with repeated administration in the same way as cocaine, and that pre exposure or concurrent exposure to CP 55,940 does not enhance sensitivity to the subsequent behavioral effects of cocaine. PMID- 9726626 TI - Effect of subchronic treatment with metrifonate and tacrine on brain cholinergic function in aged F344 rats. AB - The effects of 21-day treatment with the acetylcholinesterase inhibitors metrifonate (80 mg kg(-1) per os (p.o.)) and tacrine (3 mg kg(-1) p.o.), twice daily, on cortical and hippocampal cholinergic systems were investigated in aged rats (24-26 months). Extracellular acetylcholine levels were measured by transversal microdialysis in vivo; choline acetyltransferase and acetylcholinesterase activities were measured ex vivo by means of radiometric methods. Basal cortical and hippocampal extracellular acetylcholine levels, measured 18 h after the last metrifonate treatment, were about 15 and two folds higher, respectively, than in control and tacrine-treated rats. A challenge with metrifonate further increased cortical and hippocampal acetylcholine levels by about three and four times, respectively. Basal extracellular acetylcholine levels, measured 18 h after the last treatment with tacrine were not statistically different from those of the control rats. A challenge with tacrine increased cortical and hippocampal extracellular acetylcholine levels by about four and two times. A 75% inhibition of cholinesterase activity was found 18 h after the last metrifonate administration, while only a 15% inhibition was detectable 18 h after the last tacrine administration. The challenge with metrifonate or tacrine resulted in 90 and 80% cholinesterase inhibition, respectively. These results demonstrate that in aging rats a subchronic treatment with metrifonate results in a long-lasting, cholinesterase inhibition, and a persistent increase in acetylcholine extracellular levels which compensate for the age-associated cholinergic hypofunction. Metrifonate is therefore a potentially useful agent for the cholinergic deficit accompanying Alzheimer's disease. PMID- 9726627 TI - 5-hydroxytryptamine3 (5-HT3) receptor-mediated depolarisation of the rat isolated vagus nerve: modulation by trichloroethanol and related alcohols. AB - The ability of 2,2,2-trichloroethanol (TCE) and related alcohols to modify the 5 hydroxytryptamine3 (5-HT3) receptor-mediated depolarisation of the rat isolated cervical vagus nerve were investigated by extracellular electrophysiological recording using the 'grease gap' technique. TCE at millimolar concentrations increased the magnitude of the 5-HT3 receptor-mediated depolarisations of the rat vagus nerve by a number of agonists (5-HT, phenylbiguanide (PBG), quipazine). Concentration response curves generated for the 5-HT3 receptor agonists. 5-HT and PBG, in the absence and presence of TCE (5 mM) indicated that the potentiation in agonist-induced depolarisation was due to an increase in both agonist potency and apparent efficacy. Following apparent complete 5-HT3 receptor desensitisation (induced by either 5-HT or PBG; 100 microM for 90 min), application of TCE (5 mM) in the continued presence of either agonist induced a depolarisation of the vagus nerve. In addition to TCE, a number of related alcohols (tribromoethanol, isopentanol and 5-chloropentanol but not ethanol) at millimolar concentrations also potentiated depolarisation of the vagus nerve induced by 5-HT. Combined application of both TCE (0.1-20 mM) and isopentanol (20 mM) indicated that the potentiation of the 5-HT3 receptor-mediated depolarisation by these alcohols was not additive. The present studies indicate that the 5-HT3 receptor expressed on the cervical vagus nerve is susceptible to allosteric modulation by a number of alcohols including the anaesthetic agent TCE. Such an interaction may have relevance to the nausea and vomiting experienced by some patients following recovery from general anaesthesia. PMID- 9726628 TI - The effects of S-nitrosocaptopril on renal filtration and blood pressure in rats. AB - The present investigation was performed to evaluate the effects of S nitrosocaptopril, a novel vasodilator possessing the capacities of both an angiotensin converting enzyme inhibitor and an NO donor, on blood pressure and renal function in rats. S-nitrosocaptopril produced acute reductions in mean arterial pressure after both oral dosing (5, 10, 50 mg/kg) to chronically catheterized awake rats and intravenous administrations (0.125, 1.25, 12.5 mg/kg) to anesthetized rats. The hypotensive magnitude and duration of S nitrosocaptopril were dose-dependent. Acute pressure-associated reductions in the glomerular filtration rate and urine flow were observed only at high concentration of S-nitrosocaptopril (12.5 mg/kg, i.v.) in both awake and anesthetized rats. These decreases were transient, followed by an overshoot of glomerular filtration rate and urine flow above basal values. In contrast, captopril (i.v.) did not produce any significant acute effects on mean blood pressure and glomerular filtration rate in either awake or anesthetized rats. In rats with acute hypertension induced by NG-monomethyl-L-arginine (L-NMMA, 30 mg/kg, i.v.), S-nitrosocaptopril (0.125 mg/kg, i.v.) significantly abolished the hypertensive effects. In contrast, the hypertension was not affected by captopril. In two-kidney one-clipped Goldblatt hypertensive rats, oral administration of S-nitrosocaptopril (25 mg/kg, b.i.d.) for 10 days significantly reduced systolic blood pressure and preserved glomerular filtration rate. The oral antihypertensive effect of S-nitrosocaptopril was more potent than captopril (P < 0.05). In conclusion, these findings indicate that: (1) S-nitrosocaptopril provides both acute and chronic anti-hypertensive effects orally and intravenously, whereas captopril has only moderate chronic oral effects; and (2) S-nitrosocaptopril preferentially decreases blood pressure without markedly affecting glomerular filtration rate. PMID- 9726629 TI - Mechanisms underlying constrictor and dilator responses to perivascular nerve stimulation in canine lingual arteries. AB - In isolated canine lingual arteries denuded of the endothelium, transmural electrical stimulation (2-20 Hz) produced a frequency-related contraction which was not significantly influenced by prazosin but which was reversed to a relaxation by alpha,beta-methylene ATP. The responses were abolished by tetrodotoxin. The stimulation-induced relaxation was abolished by treatment with NG-nitro-L-arginine (L-NA, 10(-6) M) and restored by the addition of L-arginine. Neurogenic relaxation resistant to L-NA was not observed after electrical stimulation, even though the pulse width and stimulus intensity were raised. Under treatment with prazosin, alpha,beta-methylene ATP and indomethacin, the arterial strips responded to nicotine (10(-4) M) with a marked relaxation that was abolished by hexamethonium. The relaxation was significantly inhibited but not abolished by L-NA (10(-5) M), and raising the concentration of the inhibitor to 10(-4) M, did not produce additional inhibition. In the strips treated with L NA, the nicotine-induced relaxation was abolished or markedly reduced under desensitization with vasoactive intestinal peptide (VIP) or calcitonin gene related peptide (CGRP) and by treatment with high concentrations of beraprost, a stable analog of prostaglandin I2, but was unaffected by CGRP or VIP receptor antagonists. Relaxant responses to a low concentration of nicotine (5 x 10(-6) M) were abolished by L-NA and restored by L-arginine. Histochemical study demonstrated many nerve fibers and bundles containing NADPH diaphorase in the adventitia of the arteries. It is concluded that the neurogenic arterial contraction is induced mainly by ATP via stimulation of P2X purinoceptors, and that the relaxation induced by electrical stimulation or a low concentration of nicotine is mediated by nitric oxide (NO) released from perivascular nerves. In high concentrations, nicotine elicits marked relaxations possibly due to the liberation of NO from the nerve and also vasodilator substances that increase the content of cyclic AMP in the tissue. CGRP and VIP are unlikely to be involved. PMID- 9726630 TI - Protective and preventive effects of teprenone on gastric mucosal lesions in rats. AB - We have reported that neutrophil infiltration into gastric mucosa is closely related to gastric mucosal lesion development in rats with water immersion restraint stress. In this study, we examined the effect of teprenone, which is known to prevent gastric mucosal injury through stimulation of gastric mucus synthesis and secretion, on neutrophil infiltration into the gastric mucosa of rats with water immersion restraint stress. Pre- and post-administration of teprenone (200 mg/kg, p.o.) significantly attenuated the neutrophil infiltration into the gastric mucosa and the lesion development found at 6 h of water immersion restraint stress with preservation of gastric mucosal hexosamine and adherent mucus levels. These results indicate that teprenone exerts protective and preventive actions against water immersion restraint stress-induced gastric mucosal lesions in rats and suggest that these actions could be related to the preservation of gastric mucus synthesis and secretion and inhibition of neutrophil infiltration into the gastric mucosal tissue. PMID- 9726631 TI - Role of sarcoplasmic reticulum in the myorelaxant activity of nitric oxide donors in guinea pig gastric fundus. AB - The relaxant effect of two nitric oxide (NO) donors: sodium nitroprusside and 3 morpholino-sydnonimine (SIN-1) on circular smooth muscle strips isolated from guinea pig gastric fundus was studied with the view to elucidating the mechanism, which underlies the NO-induced relaxation of this tissue. Both sodium nitroprusside (10(-9)-10(-5) M) and SIN-1 (10(-9)-10(-4) M) suppressed the spontaneous fundus tone and hyperpolarized the muscle cells by about 5 mV. They antagonized the acetylcholine (10(-6) M)-induced tone and exerted their relaxant effects even when Ca2+ influx into the cells was triggered through the Na+/Ca2+ exchanger. Sodium nitroprusside and SIN-1 antagonized the contraction induced by cyclopiazonic acid (10(-5) M), a specific inhibitor of the sarcoplasmic reticulum Ca2+-ATPase. In the presence of high concentrations of sodium nitroprusside or SIN-1, cyclopiazonic acid (10(-5) M) exerted only a slight if any contractile effect. After the complete relaxation induced by sodium nitroprusside or SIN-1, the K+-channel blockers, tetraethylammonium, apamin and charybdotoxin, as well as the Ca2+ ionophore, A 23187, induced high-amplitude contractions, suggesting that the Ca2+ sensitivity of the contractile myofilaments was not affected. The results suggest that NO, released from NO donors increases the sarcoplasmic reticulum Ca2+ uptake thereby enhancing the vectorial sarcoplasmic reticulum Ca2+ release toward the plasmalemma to elicit membrane hyperpolarization and relaxation in guinea pig gastric fundus. PMID- 9726632 TI - Phosphodiesterase inhibitors suppress alpha2-adrenoceptor-mediated 5 hydroxytryptamine release from tracheae of newborn rabbits. AB - The outflow of 5-hydroxytryptamine (5-HT) from isolated tracheae of newborn rabbits was determined by high pressure liquid chromatography with electrochemical detection. This 5-HT outflow reflects release from neuroendocrine epithelial cells of the airway mucosa, as previously shown. Phenylephrine, via alpha2B-adrenoceptors, caused a transient increase in 5-HT outflow, maximally by about 250%, an effect mediated by liberation of intracellular Ca2+, as previously shown. The non-selective phosphodiesterase inhibitor 2-isobutyl-1-methylxanthine (IBMX) concentration-dependently inhibited phenylephrine-induced 5-HT release (completely at 100 microM, IC50: 1.3 microM). Likewise, benzafentrine (inhibitor of phosphodiesterase 3 and 4) and siguazodan (inhibitor of phosphodiesterase 3) also almost completely inhibited phenylephrine-induced 5-HT release with IC50 values of 1.7 and 4.2 microM, respectively. Rolipram (inhibitor of phosphodiesterase 4), in a concentration of 10 microM, which exceeds more than 10 fold the reported IC50 for phosphodiesterase 4, did not significantly affect phenylephrine-induced 5-HT release. 5-HT release induced by depolarizing concentrations of K+ (45 mM), which largely depends on extracellular Ca2+, was not affected by IBMX. In conclusion, phosphodiesterases, with characteristics of phosphodiesterase 3, appear to play an important role in the control of cyclic nucleotide mediated inhibition of 5-HT release from neuroendocrine epithelial cells. PMID- 9726633 TI - Effects of glucocorticoids on apoptosis of infiltrated eosinophils and neutrophils in rats. AB - The effects of glucocorticoids on the survival of rat eosinophils and neutrophils infiltrated into the peritoneal cavity were examined. Glucocorticoids including dexamethasone, prednisolone and hydrocortisone inhibited the survival of rat peritoneal eosinophils at 10(-6) M, whereas they prolonged survival of rat peritoneal neutrophils at 10(-8) M. Sex steroids including estradiol and progesterone did not affect cell survival. Dexamethasone decreased the viability of eosinophils after 3 days of incubation and maintained the viability of neutrophils until 4 days after incubation concentration dependently. The EC50 of dexamethasone for inhibition of the survival of eosinophils was 1.5 x 10(-8) M, and that for the spontaneous death of neutrophils was 6.4 x 10(-10) M, suggesting that glucocorticoids at concentrations that inhibit eosinophil survival prolong neutrophil survival. Analysis of DNA fragmentation of cultured eosinophils and neutrophils revealed that glucocorticoids enhance eosinophil apoptosis but inhibit neutrophil apoptosis. The effects of dexamethasone on viability and DNA fragmentation were counteracted by the glucocorticoid receptor antagonist, mifepristone, concentration dependently. These findings indicate that glucocorticoids induce contradictory effects via the glucocorticoid receptor on rat eosinophils and neutrophils extravasated to an inflammatory locus such as the peritoneal cavity by modulating apoptosis. PMID- 9726634 TI - Action of chloroquine on nitric oxide production and parasite killing by macrophages. AB - Chloroquine is known to inhibit several functions of macrophages, but its effect on the nitric oxide (NO)-dependent parasite killing capacity of macrophages has not been documented. NO synthesis by interferon-gamma-induced mouse and casein elicited rat macrophages was significantly and irreversibly inhibited by chloroquine. The activity of the inducible NO synthase was not directly altered, but previous incubation of macrophages with chloroquine decreased it. Chloroquine did not alter arginase activity or arginine uptake. NADPH diaphorase activity, an indicator of NO synthase was impaired. Western blotting showed that inducible NO synthase synthesis was blocked by chloroquine. The blocking of NO formation by chloroquine resulted in increased infection of mouse peritoneal macrophages by Trypanosoma cruzi (T. cruzi). This suggests that chloroquine decreases NO formation by macrophages by inhibiting the induction of NO synthase. The findings are further evidence that NO is involved in the anti-parasitic response of macrophages. PMID- 9726635 TI - Pharmacological properties of the GABA(A) receptor complex from brain regions of (hypoemotional) Roman high- and (hyperemotional) low-avoidance rats. AB - The pharmacological properties of benzodiazepine binding sites of the gamma aminobutyric acid (GABA)A receptor complex from cortical, hippocampal and cerebellar membranes of Roman high-avoidance (RHA/Verh) and Roman low-avoidance (RLH/Verh) rats were investigated. No major differences between the two lines were found in the binding parameters of [3H]flunitrazepam (a non-selective agonist). [3 H]zolpidem (a Type I selective agonist) or [3 H]ethyl 8-azido-6 dihydro-5-methyl-6-oxo-4H-imidazol[1,5-a]-[1,4]benzodiazepine- 3-carboxylate (Ro15-4513) (a partial inverse agonist). Neither the Kd values nor the Bmax for these ligands differed between RHA/Verh and RLA/Verh rats in any of the brain regions studied. As a result, the proportion of Type I binding sites in cortical and hippocampal membranes of RHA/Verh and RLA/Verh rats or the 'diazepam sensitive' and the 'diazepam-insensitive' binding sites in cerebellar membranes, calculated from the [3H]flunitrazepam and [3H]zolpidem maximal binding sites or from [3H]Ro15-4513 binding (in the absence or in presence of diazepam), respectively, was also similar. Furthermore, there were no differences between the two rat lines in the allosteric interactions between GABA and the benzodiazepine binding sites (labeled with [3H]flunitrazepam) in all three areas tested or the Type I binding sites (labeled with [3H]zolpidem) in the hippocampus. In contrast, RLA/Verh rats showed a significant reduction in the allosteric interactions between GABA and [3H]zolpidem binding sites in the cortex. As a whole, these results indicate the absence of generalized between line differences in the GABA(A) receptor complex showing, at the same time, the existence of some specific differences in allosterism within the GABA(A) complex. These differences may contribute to the divergent emotional responses which characterize the RHA/Verh and RLA/Verh rat lines. PMID- 9726636 TI - Effect of the protein kinase inhibitors, 1-(5-isoquinolinylsulfonyl)-2 methylpiperazine H-7 and N-(2-[methylamino]ethyl)-5-isoquinoline-sulfonamide H-8 on Lewis lung carcinoma tumor progression. AB - The effects of 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine H-7 (a cAMP dependent protein kinase and protein kinase C inhibitor), n-(2 [methylamino]ethyl)-5-isoquinoline-sulfonamide H-8 (a cAMP- and cGMP-dependent protein kinase inhibitor) and indomethacin (IND, a cyclooxygenase inhibitor) on both the spontaneous metastatic ability of 3LL (Lewis lung carcinoma) tumor cells and anti-tumor host response were studied. The study of tumor progression showed that H-7 and H-8 (2 mg kg(-1) day(-1) , i.p., for 8 days) significantly reduced the mean number of metastases (0.8 +/- 0.2 and 1.0 +/- 0.7, respectively, P < 0.05) with respect to the number of lung metastases (4.2 +/- 2.1) observed in the control group. In turn, the highest tumor-specific cytotoxicity response (50% increase vs. non-treated target cells) was observed when both animal and tumor cells were treated with H-8. This suggests that the protein kinase inhibitors could inhibit tumor progression toward lung metastases formation by blocking the immunosuppressor mechanism triggered by agents that increase intracellular cAMP. PMID- 9726637 TI - Interaction of lipophilic VIP derivatives with recombinant VIP1/PACAP and VIP2/PACAP receptors. AB - Stearyl vasoactive intestinal polypeptide has been reported to be a VIP (vasoactive intestinal polypeptide) receptor agonist of high potency with an original bioavailability and action. We synthesized three fatty acyl derivatives, myristyl-, palmityl- and stearyl-[Nle17]VIP, and tested their capacity to recognize recombinant rat- and human VIP1- and VIP2/PACAP (pituitary adenylate cyclase-activating polypeptide) receptors and to stimulate adenylate cyclase activity. The three lipophilic analogues bound with high affinity (from 0.5 to 20 nM) to both receptor subtypes but did not distinguish between them. In preparations expressing a high density of human VIP1/PACAP receptors, the three lipophilic analogues had the same efficacy as VIP and [Nle17]VIP. In preparations expressing the rat receptors, stearyl-[Nle17]VIP had a lower efficacy than the other peptides tested. In preparations expressing a low level of VIP1/PACAP receptors and in those expressing VIP2/PACAP receptors, all analogues behaved like partial agonists. The lowest efficacy was observed for stearyl-[Nle17]VIP on the VIP2/PACAP receptor subclass. Based on our results, a complex pattern of in vivo biological effects of the lipophilic VIP derivatives should be expected: these compounds might behave as full agonists, partial agonists, or antagonists of the VIP response, depending on the number and the subtype of receptor expressed. PMID- 9726638 TI - Discrimination between plasma membrane and intracellular target sites of sphingosylphosphorylcholine. AB - On the background of the emerging concept of G protein-coupled sphingolipid receptors, Ca2+ mobilization by sphingosylphosphorylcholine (SPPC) in intact cells and SPPC-induced Ca2+ release in permeabilized cells, both occurring at similar, micromolar concentrations, were characterized and compared. In intact human embryonic kidney (HEK-293) cells, SPPC rapidly increased [Ca2+]i by mobilization of Ca2+ from thapsigargin-sensitive stores. In saponin-permeabilized HEK-293 cells, SPPC released stored Ca2+, in a manner similar to but independent of inositol 1,4,5-trisphosphate. Only the action of SPPC on intact cells, but not that in permeabilized cells, was, at least in part, sensitive to pertussis toxin. In addition and most important, Ca2+ release by SPPC in permeabilized cells was not stereoselective, whereas in intact cells only the naturally occurring D erythro-SPPC, but not L-threo-SPPC, increased [Ca2+]i. Stereoselectivity of SPPC induced [Ca2+]i increase was also demonstrated in bovine aortic endothelial cells. In conclusion, Ca2+ mobilization by SPPC in intact cells is independent of the previously described SPPC-gated Ca2+ channel on endoplasmic reticulum but probably mediated by a membrane sphingolipid receptor. Thus, SPPC can regulate Ca2+ homeostasis by acting apparently at two cellular targets, which exhibit clearly distinct recognition patterns. PMID- 9726639 TI - Synthesis of dopamine from L-3,4-dihydroxyphenylalanine by human amniotic epithelial cells. AB - In this study, the ability of human amniotic epithelial cells to synthesize dopamine from L-3,4-dihydroxyphenylalanine (L-DOPA) was examined. Dopamine synthesis was significantly increased time and L-DOPA concentration dependently, suggesting the presence of an aromatic L-amino acid decarboxylase enzyme. This was confirmed by the decrease in dopamine synthesis in the presence of an aromatic L-amino acid decarboxylase inhibitor, benserazide. These findings suggest that human amniotic epithelial cells have the capacity to take up and convert L-DOPA into dopamine. PMID- 9726640 TI - The delta1-opioid receptor antagonist, 7-(benzospiroindanyl)naltrexone [correction of 7-benzylspiroindanylnaltrexone], prolongs renal allograft survival in a rat model. AB - In this study we demonstrate allograft survival in a rat model of renal transplantation using the delta1-opioid receptor antagonist, 7 (benzospiroindanyl)naltrexone [corrected]. Treatment with 7 (benzospiroindanyl)naltrexone [corrected] caused 50% of the rats to survive longer than 100 days (untreated, 11 +/- 3 days). Naltrindole, a delta-opioid receptor antagonist without subtype selectivity, also promoted graft survival but was substantially less effective, suggesting that antagonism at delta1-opioid receptors is involved in allograft survival. PMID- 9726642 TI - Asp10 in Lys-gamma2-MSH determines selective activation of the melanocortin MC3 receptor. AB - The melanocortin MC3 and MC4 receptors are the main melanocortin receptors expressed in brain. Of the endogenous melanocortins, gamma2-melanocortin stimulating hormone (MSH) selectively activates the melanocortin MC3 receptor, whereas alpha- and beta-MSH activate all melanocortin receptors. The aim was to gain an insight into the contribution of amino acids in positions 5 and 10 of melanocortins to the selectivity of [Nle4]Lys-gamma2-MSH for the melanocortin MC3 receptor versus the melanocortin MC4 receptor. Introduction of Asp10 into [Nle4]alpha-MSH as in [Nle4,Gly5,Asp10]alpha-MSH selectively increased the EC50 value for the melanocortin MC4 receptor. Conversely, removal of Asp10, as in [Nle4,Gly10]Lys-gamma2-MSH, selectively decreased the EC50 value for the melanocortin MC4 receptor. Thus, Asp10 in Lys-gamma2-MSH determined selectivity for the melanocortin MC3 receptor versus the melanocortin MC4 receptor. PMID- 9726641 TI - Glucocorticoids inhibit the bradykinin B2 receptor increase induced by interleukin-1beta in human bronchial smooth muscle cells. AB - We studied the effect of the glucocorticoids, dexamethasone and budesonide, on the interleukin-1beta-induced increase of bradykinin B2 receptors in cultured human bronchial smooth muscle cells, a cellular model of bronchial hyperreactivity. Both compounds prevented the increase of the bradykinin B2 mRNA and the bradykinin-induced inositol phosphate accumulation. These results demonstrate a direct effect of glucocorticoids on airway smooth muscle hyperresponsiveness mediated through inhibition of the over-expression of receptors for contractile mediators induced by inflammatory mediators. PMID- 9726643 TI - Topical ondansetron attenuates nociceptive and inflammatory effects of intradermal capsaicin in humans. AB - Topical application of the 5-HT3 receptor antagonist ondansetron (50-250 microg) delivered in a pluronic lecithin organogel vehicle (PLO, 0.5 ml) produced dose dependent attenuation of nociceptive and inflammatory effects of intradermally injected capsaicin (10 microg/10 microl) in humans. Significant, dose-dependent analgesic effects were produced by 100 microg and 250 microg doses of ondansetron; these doses also reduced mechanical hyperalgesia produced by capsaicin. However, only 250 microg dose of ondansetron diminished capsaicin induced inflammatory flare. PMID- 9726644 TI - GR205171 blocks apomorphine and amphetamine-induced conditioned taste aversions. AB - The tachykinin NK1 receptor antagonist, GR205171 ([2-methoxy-5-(5-trifluoromethyl tetrazol-1-yl)-benzyl]-(2S-phenyl -piperidin-3S-yl)-amine), is a potent inhibitor of emesis induced by a wide variety of emetogens. This is in contrast to 5-HT3 (5 hydroxytryptamine3) receptor antagonists, such as ondansetron, which have a more restricted antiemetic profile. The present study evaluated the efficacy of GR205171, in comparison with ondansetron to block the acquisition of a conditioned taste aversion induced by either apomorphine (0.25 mg kg(-1) s.c.) or by amphetamine (0.5 mg kg(-1) s.c.) in rats. Pretreatment with GR205171 (0.1-1.0 mg kg(-1) s.c.) and ondansetron (0.001-0.1 mg kg(-1) s.c.) produced a dose dependent blockade of conditioned taste aversions evoked by apomorphine. In contrast, the acquisition of conditioned taste aversions induced by amphetamine was inhibited by GR205171 (0.3-0.5 mg kg(-1) s.c.), but only attenuated by ondansetron (0.001-0.1 mg kg(-1) s.c.). These results suggest that tachykinin NK1 receptor antagonists may have potential in the treatment of drug-induced conditioned aversive behaviour and nausea. PMID- 9726645 TI - Disruption of dopamine D1 receptor gene expression attenuates alcohol-seeking behavior. AB - The role of the dopamine D1 receptor subtype in alcohol-seeking behaviors was studied in mice genetically deficient in dopamine D1 receptors (D1 -/-). In two tube free choice limited (1-5 h) and continuous (24 h) access paradigms, mice were exposed to water and increasing concentrations of ethanol (3%, 6% and 12% w/v). Voluntary ethanol consumption and preference over water were markedly reduced in D1 -/- mice as compared to heterozygous (D1 +/-) and wild-type (D1 +/+) controls, whereas overall fluid consumption was comparable. When offered a single drinking tube containing alcohol as their only source of fluid for 24 h, D1 -/- mice continued to drink significantly less alcohol than D1 +/+ and D1 +/- mice. Dopamine D2 receptor blockade with sulpiride caused a small but significant reduction in alcohol intake and preference in D1 +/+ mice and attenuated residual alcohol drinking in D1 -/- mice. Dopamine D1 receptor blockade with SCH-23390 very effectively reduced alcohol intake in D1 +/+ and D1 +/- mice to the level seen in untreated D1 -/- mice. These findings suggest involvement of both dopamine D1 and D2 receptor mechanisms in alcohol-seeking behavior in mice; however, these implicate D1 receptors as having a more important role in the motivation for alcohol consumption. PMID- 9726646 TI - In vivo and in vitro evaluation of AMPA receptor antagonists in rat hippocampal neurones and cultured mouse cortical neurones. AB - The effects of four glutamate receptor antagonists on alpha-amino-3-hydroxy-5 methylisoxazole-4-propionic acid (AMPA)- and N-methyl-D-aspartate (NMDA) responses were evaluated using both in vitro and in vivo electrophysiological techniques: whole cell patch-clamp recordings from cultured mouse cortical neurones and microiontophoresis in the rat hippocampus. The compounds tested were NBQX (2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo(F)quinoxaline), GYKI 52466 (1-(4 amino-phenyl)-4-methyl-7,8-methyl-endioxyl-5H-2,3-benzodiaze pine), PNQX (pyrido[3, 4-f]quinoxaline-2,3-dione, 1,4,7,8,9,10-hexahydro-9-methyl-6-nitro-, methanesulfonate), NS377 (7-ethyl-5-phenyl-1,6,7,8-tetrahydro-1,7-diaza-as indacene-2 ,3-dione), and MK-801 ((+)-5-methyl-10,11-dihydro-5H dibenz(a,d)cycloheptene-5,10-imine hydrogen maleate). In vitro, the IC50 values (in microM) for inhibition of AMPA-evoked inward currents were approximately 0.4 for NBQX, approximately 7.5 for GYKI 52466, approximately 1 for PNQX and approximately 15 for NS377. PNQX and NS377 also inhibited NMDA-induced currents with IC50 values at approximately 5 and approximately 18 microM, respectively, while NBQX at 60 microM and GYKI 52466 at 100 microM had only weak effects. The ED50 values in micromol/kg i.v. for inhibition of AMPA-evoked hippocampal neuronal spike activity in vivo were approximately 32 for NBQX, approximately 19 for GYKI 52466, approximately 17 for PNQX and approximately 11 for NS377 with efficacy values (maximal inhibition) between 71% and 81%. The ED50 values (in [Lmol/kg i.v.) and efficacy values for inhibition of NMDA-evoked hippocampal neuronal spike activity were approximately 28 with an efficacy of 61% for NBQX, approximately 16 with 35% for PNQX and approximately 6 with 61% for NS377. GYKI 52466 did not significantly affect NMDA responses, whereas MK-801 showed NMDA specificity in vivo. PMID- 9726647 TI - The role of BK(Ca) channels in the nitric oxide-mediated regulation of adrenal catecholamine secretion. AB - We examined whether high conductance Ca2+-activated K+ (BK(Ca)) channels are involved in the modulatory action of nitric oxide (NO) on the secretion of adrenal catecholamines in response to splanchnic nerve stimulation and acetylcholine in anesthetized dogs. The NO donor 3-(2-hydroxy-1-methyl-2 nitrosohydrazino)-N-methyl-1-propanamin e (NOC 7), the BK(Ca) channel blocker charybdotoxin and acetylcholine were administered intraarterially (i.a.) into the adrenal gland. NOC 7 infusion (2 microg min(-1)) inhibited increases in catecholamine output induced by splanchnic nerve stimulation (1-3 Hz) and acetylcholine (0.75-3 microg). Charybdotoxin infusion (100 ng min(-1)) did not affect increases in catecholamine output induced by splanchnic nerve stimulation and acetylcholine. Charybdotoxin blocked the NOC 7-induced inhibition of increases in catecholamine output induced by splanchnic nerve stimulation but not by acetylcholine. These results suggest that NO may inhibit the secretion of adrenal catecholamines induced by splanchnic nerve stimulation through activation of BK(Ca) channels. PMID- 9726648 TI - Electrical but not chemical kindling increases sensitivity to some phencyclidine like behavioral effects induced by the competitive NMDA receptor antagonist D CPPene in rats. AB - We have previously reported that a competitive N-methyl-D-aspartate (NMDA) receptor antagonist, DL-[E]-2-amino-4-methyl-5-phosphono-3-pentenoic acid (CGP 37849), produces stereotyped behaviors and hyperlocomotion in amygdala kindled rats at doses which do not induce such phencyclidine (PCP)-like behaviors in nonkindled rats, indicating that kindling predisposes rats to such adverse effects of competitive NMDA receptor antagonists. From these data we predicted that epileptic patients may exhibit a hypersensitivity to PCP-like adverse effects of competitive NMDA receptor antagonists, which was subsequently confirmed in a clinical trial with D-CPPene (SDZ EAA-494; 3-(2-carboxypiperazine 4-yl)propenyl-1-phosphonate). For further exploration of the functional alterations in NMDA receptor responsiveness produced by kindling, we studied whether the enhanced susceptibility of amygdala-kindled rats to PCP-like adverse effects of CGP 37849 is also observed with D-CPPene. Furthermore, we determined whether the enhanced susceptibility of kindled rats to such adverse effects occurs only after relatively short intervals following the last seizure, as used in our previous study, or is a more permanent phenomenon. For this purpose, we compared adverse effects in kindled rats not only with naive (non-implanted) controls, as done in our previous study, but used electrode-implanted nonkindled rats as an additional control to assess the possible bias of mere electrode implantation. In addition, we studied whether the enhanced susceptibility to NMDA receptor antagonists of electrically kindled rats is also present in chemically kindled animals. In some experiments, the PCP-like uncompetitive NMDA receptor antagonist MK-801 (dizocilpine) was included for comparison. In amygdala kindled rats, D-CPPene produced significantly more stereotyped behaviors than in electrode-implanted or naive nonkindled controls. The enhanced sensitivity of electrically kindled rats to PCP-like stereotypies induced by D-CPPene was observed both 7 and 180 days after the last kindled seizure, indicating a long lasting if not permanent hypersensitivity to these adverse effects. In addition, more intense circling was observed in amygdala kindled rats, whereas hyperlocomotion only tended to be more intense after D-CPPene in kindled rats. These alterations in D-CPPene-induced behaviors were not observed after chemical kindling with pentylenetetrazole, but D-CPPene induced significantly less hypothermia in chemically kindled rats both 7 and 70 days after the last seizure. The data demonstrate that kindling produces long-lasting alterations in some adverse effects of D-CPPene, substantiating that epileptogenesis as initiated by kindling renders the brain more susceptible to PCP-like behavioral side effects of competitive NMDA receptor antagonists. PMID- 9726649 TI - Evidence for a unique profile of levetiracetam in rodent models of seizures and epilepsy. AB - The protective and adverse effect potentials of levetiracetam ((S)-alpha-ethyl-2 oxo-pyrrolidine acetamide) in rodent models of seizures and epilepsy were compared with the profile of several currently prescribed and newly developed antiepileptic drugs. Levetiracetam was devoid of anticonvulsant activity in the acute maximal electroshock seizure test and in the maximal pentylenetetrazol seizure test in mice (up to 540 mg/kg, i.p.) but exhibited potent protection against generalised epileptic seizures in electrically and pentylenetetrazol kindled mice (ED50 values = 7 and 36 mg/kg, respectively, i.p.). This differs markedly from established and most new antiepileptic drugs which induce significant protection in both the acute seizure tests and the kindling models. Furthermore, levetiracetam was devoid of anticonvulsant activity in several maximal chemoconvulsive seizure tests although an interesting exception was the potent protection observed against secondarily generalised activity from focal seizures induced by pilocarpine in mice (ED50 value = 7 mg/kg, i.p.), pilocarpine and kainic acid in rats (minimum active dose = 17 and 54 mg/kg, respectively, i.p.). The protection afforded by levetiracetam on the threshold for methyl-6,7 dimethoxy-4-ethyl-beta-carboline-3-carboxylate (DMCM)-induced seizures persisted after chronic administration (17-170 mg/kg, i.p., twice daily/14 days) and levetiracetam did not lower the seizure threshold for the proconvulsant action of the inverse benzodiazepine receptor agonist, N-methyl-beta-carboline-3 carboxamide (FG 7142). The main metabolite of levetiracetam (ucb L057; (S)-alpha ethyl-2-oxo-1-pyrrolidine acetic acid) was found to be inactive in sound sensitive mice after acute administration of doses up to 548 mg/kg, i.p. Levetiracetam induced only minor behavioural alterations in both normal and amygdala-kindled rats (54-1700 mg/kg, i.p.) resulting in an unusually high safety margin between rotarod impairment and seizure suppression of 148 in corneally kindled mice and 235 in Genetic Absence Epilepsy Rats from Strasbourg. In comparison, existing antiepileptic drugs have ratios between 2 and 17 in the corneally kindled mouse model. These studies reveal a unique profile of levetiracetam in rodent models. Characteristics are a general lack of anticonvulsant activity against maximal, acute seizures and selective protection with a very high safety margin in genetic and kindled animals and against chemoconvulsants producing partial epileptic seizures. This activity differs markedly from that of the established and newly introduced antiepileptic drugs and appears to derive from the parent compound since its major metabolite was inactive in all models studied. Together these results therefore suggest that levetiracetam may offer an effective, broad-spectrum treatment of epileptic seizures in patients, with a minimum of adverse effects. PMID- 9726650 TI - Flesinoxan treatment reduces 5-HT1A receptor mRNA in the dentate gyrus independently of high plasma corticosterone levels. AB - Flesinoxan acts as a full 5-HT1A receptor agonist and displays anxiolytic and anti-depressant properties. 5-HT1A receptor agonists, including flesinoxan, increase corticosterone (B) levels in the blood and reduces 5-HT1A receptor mRNA expression in the hippocampus. In this study, we examined whether the 5-HT1A receptor downregulation induced by flesinoxan involves corticosterone control of 5-HT1A receptor gene transcription. In experiment I, intact male Wistar rats (180 200 g) were treated with flesinoxan (1.0, 3.0 and 10 mg/kg bw, sc) or vehicle and decapitated 3 h later. Flesinoxan administration resulted in a significant, dose dependent downregulation of 5-HT1A receptor mRNA in the dentate gyrus and dorsal raphe nucleus. In experiment II, rats were sham-operated and implanted with a cholesterol pellet (100 mg) or were adrenalectomized and implanted with a corticosterone pellet (20 mg corticosterone + 80 mg cholesterol). Flesinoxan injection also caused a dose-dependent decrease of 5-HT1A mRNA in the dentate gyrus of adrenalectomized animals with corticosterone replacement. There was no effect in the dorsal raphe nucleus. In experiment III, adrenalectomized and adrenalectomized + corticosterone rats were sc injected with flesinoxan (10 mg/kg bw) or vehicle, and flesinoxan appeared to downregulate 5-HT1A receptor expression in the dentate gyrus independently of corticosterone as well. No significant effects were observed in the dorsal raphe nucleus. It is concluded that flesinoxan reduces 5-HT1A receptor expression in the dentate gyrus both through homologous downregulation and a corticosterone-mediated effect on the serotonergic (5-HT) system. PMID- 9726651 TI - Validation of S-1'-[18F]fluorocarazolol for in vivo imaging and quantification of cerebral beta-adrenoceptors. AB - S-1'-[18F]fluorocarazolol (S-(-)-4-(2-hydroxy-3-(1'-[18F]fluoroisopropyl) aminopropoxy)carba zole, a non-subtype-selective beta-adrenoceptor antagonist) has been investigated for in vivo studies of beta-adrenoceptors. Previous results indicated that uptake of this radioligand in heart and lung can be inhibited by beta-adrenoceptor agonists and antagonists. In the present study, blocking, displacement and saturation experiments were performed in rats, in combination with metabolite analysis to investigate the suitability of this radioligand for in vivo positron emission tomography (PET) imaging and quantification of beta adrenoceptors in the brain. The results demonstrate that, (i) the uptake of S-1' [18F]fluorocarazolol reflects specific binding to beta-adrenoceptors, (ii) binding of S-1'-[18F]fluorocarazolol to atypical or non-beta-adrenergic sites is negligible, (iii) uptake of radioactive metabolites in the brain is less than 25% of total radioactivity, 60 min after injection, (iv) in vivo measurements of receptor densities (Bmax) in cortex, cerebellum, heart, lung and erythrocytes are within range of densities determined from in vitro assays, (v) binding of S-1' [18F]fluorocarazolol can be displaced. In conclusion, S-1'-[18F]fluorocarazolol seems to possess the appropriate characteristics to visualize and quantify beta adrenoceptors in vivo in the central nervous system using PET. PMID- 9726652 TI - Involvement of spinal dopamine receptors in mediation of the hypotensive and bradycardic effects of systemic quinpirole in anaesthetised rats. AB - This study examined the involvement of spinal dopamine D2 receptors in the cardiovascular effects induced by intravenous administration of the selective dopamine D2 receptor agonist quinpirole, as has been previously reported for the hypotensive action of systemic bromocriptine. In normotensive pentobartitone anaesthetised rats, intravenous injection of quinpirole (25 to 1000 microg/kg) decreased mean aortic pressure and heart rate in a dose-related manner. The intravenous (0.5 mg/kg) or intrathecal (40 microg/rat at T9-T10) pretreatment with domperidone, a dopamine D2 receptor antagonist that does not cross the blood brain barrier, significantly reduced the maximal hypotensive and bradycardic responses to intravenous quinpirole (1000 microg/kg). In contrast, the latter effects were fully abolished either by intravenous metoclopramide (5 mg/kg) or combined pretreatment with intravenous and intrathecal domperidone. In addition, when injected intrathecally at the T9-T10 level of the spinal cord, quinpirole (7.7 to 61.4 microg/rat) also produced dose-dependent depressor and bradycardic effects which could be blocked by intrathecal, but not intravenous, domperidone pretreatment. This suggests that, in anaesthetised normotensive rats, the hypotensive and bradycardic responses to intravenous quinpirole are fully mediated by dopamine D2 receptors, some of which are located in the peripheral circulation and some of which are located within the spinal cord. The latter finding is novel, suggesting that partial spinal mediation may not be peculiar to bromocriptine, as was previously thought. Rather, partial spinal mediation may be common to most dopamine D2 receptor agonists. PMID- 9726653 TI - Vasorelaxant effect of olprinone, an inhibitor of phosphodiesterase 3, on mesenteric small artery and vein of rabbits. AB - The effects of olprinone, a cardiotonic agent that inhibits cyclic GMP (cGMP) inhibited phosphodiesterase, was studied on isolated rabbit mesenteric small artery and vein. In the presence of indomethacin and propranolol, olprinone at concentrations of 10 nM to 10 microM and 1 microM to 100 microM relaxed norepinephrine-stimulated mesenteric artery and vein in a concentration-dependent manner, respectively. The relaxation was not endothelium-dependent in the artery. Removal of the endothelium, however, increased marginally the response of the vein to olprinone. Olprinone-induced relaxation was less pronounced in arteries contracted with high KCl solution + norepinephrine than in those contracted with norepinephrine alone. Nicardipine inhibited this attenuating effect of high KCl solution on the olprinone-induced relaxation. Olprinone (1 microM) enhanced the relaxation of artery and vein in response to a cAMP-increasing agent, 6-(3 dimethylaminopropionyl) forskolin (NKH477), but not to a cGMP- increasing agent, glyceryl trinitrate. Norepinephrine (10 microM) and caffeine (5 mM) elicited a transient, phasic contraction of the artery in Ca2+-free solution. Both olprinone and NKH477 attenuated more potently the norepinephrine-induced contraction than the caffeine-induced contraction. When norepinephrine (10 microM) and caffeine (5 mM) were successively applied in Ca2+-free solution, the contractile effect of caffeine was diminished compared to that in artery which had not been pretreated with norepinephrine. When the contraction in response to norepinephrine was partially attenuated by 1 microM olprinone, the following contraction evoked by caffeine was enlarged. It is concluded that olprinone relaxes the small artery more strongly than the vein via its direct action on smooth muscles. It is suggested that olprinone attenuates norepinephrine-induced contraction through inhibition of receptor-operated transmembrane Ca2+ influx and Ca2+ release from intracellular storage sites. PMID- 9726654 TI - A role of myofilament Ca2+ sensitivity in enhanced vascular reactivity in cardiomyopathic hamsters. AB - We compared the contractile responses to vasoconstrictors in aortas from 20- to 22-week old cardiomyopathic hamsters, BIO 53.58 strain, and age-matched F1b strain controls. Aortas from cardiomyopathic hamsters exhibited greater contractions in response to phenylephrine, angiotensin II, and high K+ than did the controls. Neither endothelium removal nor the presence of indomethacin and N(omega)-nitro-L-arginine (L-NNA) affected the enhanced contractile responses to these vasoconstrictors, indicating no involvement of endogenous prostanoids and nitric oxide from the endothelium. The contractile response to phorbol-12,13 dibutyrate (PDB) was also more markedly increased in cardiomyopathic aortas regardless of whether extracellular Ca2+ was present. The contractile response of cardiomyopathic aorta to phenylephrine was more sensitive to the inhibitory actions of the protein kinase C inhibitors staurosporine and calphostin C than was that of control aorta. These results suggest that activation of protein kinase C is partly involved in the enhanced phenylephrine response of cardiomyopathic aorta. None of nifedipine, ryanodine, and cyclopiazonic acid modified the maximum contractions induced by phenylephrine in either cardiomyopathic aortas or controls. The Ca2+ sensitivity of tension was significantly increased in beta-escin-skinned smooth muscle of mesenteric artery from cardiomyopathic hamsters compared to that of controls. PDB induced Ca2+ sensitization, but significantly only in cardiomyopathic hamsters. We propose that the enhanced vascular reactivity in cardiomyopathic hamsters may primarily result from increased Ca2+ sensitivity of contractile proteins. In addition, protein kinase C-mediated Ca2+ sensitization may further contribute to the enhanced vascular response to agonists. PMID- 9726655 TI - Role for adenosine A1 and A2 receptors in femoral vasodilatation induced by intra arterial adenosine in rabbits. AB - The vasodilator effects of adenosine injected into the femoral artery (i.a.) of rabbits were analyzed. Single bolus i.a. doses of adenosine (0.3-10 microg) and 5'-(N-cyclopropyl)-carboxamidoadenosine (CPCA) (0.03-1 microg), an adenosine A2 receptor agonist, produced dose-dependent increases in femoral blood flow and decreases in resistance, almost without affecting blood pressure, heart rate, left ventricular (LV) pressure, and LVd P/dt max, even though CPCA elicited slight decreases in arterial blood pressure and LV pressure. On the other hand, bolus i.a. injections of N6-cyclopentyladenosine (CPA) (1-30 microg), an adenosine A1 receptor agonist, caused a relatively weak increase in blood flow, but markedly affected cardiac parameters, especially heart rate and LVd P/dt max. I.v. treatment with 3,7-dimethyl-1-propargylxanthine (DMPX)(2 mg kg(-1)), an antagonist of adenosine A2 receptors, or 8-phenyltheophylline (1 mg kg(-1)), an antagonist of adenosine A1 receptors, significantly attenuated the vasodilator response to adenosine, but not that to acetylcholine. Decreases in blood pressure, heart rate, LV pressure, LVdP/dt max and femoral vascular resistance, and increases in the blood flow elicited by CPA were not significantly modified by the DMPX treatment, but when this was combined with 8-phenyltheophylline, the responses to CPA were completely abolished. The present results indicate that the adenosine-induced femoral vasodilatation in rabbits may be mediated throughout activation of both adenosine A1 and A2 receptors. PMID- 9726656 TI - The effect of nociceptin, an endogenous ligand for the ORL1 receptor, on rat colonic contraction and transit. AB - To assess the role of ORL1 (opioid receptor-like 1) receptor in the bowel movement, we investigated the effect of nociceptin on colonic contraction and transit in rats. Nociceptin (0.1-100 nM) concentration-dependently caused an immediate tonic contraction followed by rhythmic waves of contractions in the isolated colon. The response to nociceptin (10 nM) was not affected by the classical opioid receptor antagonists, naloxone, naltrindole and nor binaltorphimine. Suppression of effect of inhibitory neurotransmitters using pituitary adenylate cyclase activating polypeptide(6-38) (PACAP-(6-38); 3 microM), vasoactive intestinal polypeptide(10-28) (VIP-(10-28); 3 microM) and N(omega)-nitro-L-arginine methyl ester (L-NAME; 100 microM) did not influence the nociceptin-induced contractions. In anesthetized rats, intravenous administration of nociceptin (1 microg/kg) or morphine (1 mg/kg) caused phasic contractions in the proximal colon. Pretreatment with naloxone (300 microg/kg, i.v.) abolished the contractions induced by morphine, but not by nociceptin. The rate of large intestinal transit was dose-dependently accelerated by nociceptin (0.03-3 microg/kg, s.c.), but was retarded by morphine (1.7-5 mg/kg, s.c.). These results indicate that stimulation of ORL1 receptor accelerates the colonic contraction and transit independently from opioid receptors. PMID- 9726657 TI - ONO-5046 is a potent inhibitor of neutrophil elastase in human pleural effusion after lobectomy. AB - The imbalance of neutrophil elastase and alpha1-antitrypsin in pleural effusion after lobectomy and the effects of the neutrophil elastase inhibitors, sodium N [2-[4-(2,2-Dimethylpropionyloxy)phenyl-sulfonylamino]benzo yl]aminoacetic acid (ONO-5046) and purified alpha1-antitrypsin, on neutrophil elastase activity were determined. The amount of neutrophil elastase complexed to alpha1-antitrypsin, measured by an enzyme-linked immunosorbent assay, was 170 times higher in pleural effusion than in blood 3 h after lobectomy. The alpha1-antitrypsin levels measured by laser nephelometry did not increase in either blood or pleural effusion. Although neutrophil elastase activity, measured by the hydrolysis of succinyl-(Ala)3-p-nitroanilide, was not detected in blood, it was increased in pleural effusion 3 h and 24 h after lobectomy. ONO-5046, but not alpha1 antitrypsin, reduced the neutrophil elastase activity in pleural effusion. There is an imbalance of neutrophil elastase and alpha1-antitrypsin in pleural effusion after lobectomy. ONO-5046 is a potent inhibitor of neutrophil elastase activity in human pleural effusion. PMID- 9726658 TI - In vitro inhibition of human colonic motility with SR 59119A and SR 59104A: evidence of a beta3-adrenoceptor-mediated effect. AB - The new beta3-adrenoceptor is present in the gastrointestinal tract of various species. This study aimed to show that this receptor modulates human colonic motility in vitro. We used circular muscle strips from the human colon suspended in single organ baths containing Krebs solution and subjected to an initial 1.5-2 g tension. We measured the effects of different beta3-adrenoceptor agonists, including SR 59104A (N-[(6-hydroxy-1,2,3,4-tetrahydronaphthalen-(2R)-2-yl)methyl] (2 R)-2-hydroxy-2-(3-chlorophenyl)ethanamine hydrochloride), SR 59119A (N-[(7 methoxy-1,2,3,4-tetrahydronaphthalen-(2R)-2-yl)methyl]-(2R) -2-hydroxy-2-(3 chlorophenyl)ethanamine hydrochloride), BRL 37344 (R,R + S,S) [4-[2-[[2-(3 chlorophenyl)-2-hydroxyethyl]-amino] propyl] phenoxy] acetic acid), and of isoprenaline and salbutamol in the absence or in the presence of propranolol alone or in combination with the beta3-adrenoceptor antagonist SR 59230A (3-(2 ethylphenoxy)-1-[(1S)-1,2,3,4-tetrahydro-naphthalen-1- ylamino]-(2S)-2-propanol oxalate) on amplitude of spontaneous contractions. To evaluate a possible beta2 adrenoceptor-mediated effect, we studied the action of these compounds on human isolated bronchi. On the human isolated colon, SR 59119A, SR 59104A and isoprenaline reduced the initial amplitude of spontaneous contractions by 60%. The curves obtained in the presence of antagonists suggested an action mediated by beta3-adrenoceptor stimulation, since propranolol did not antagonize the action of SR 59119A and SR 59104A, whereas the combination of propranolol and SR 59230A significantly displaced the concentration-response curve of these agonists to the right. This study provides pharmacological evidence of modulation of human colonic motility, and especially of the amplitude of spontaneous contractions, by the atypical beta-adrenoceptor, the beta3-adrenoceptor. PMID- 9726659 TI - The response of rat colonic mucosa to 5-hydroxytryptamine in health and following restraint stress. AB - The present study characterized the rat colonic secretory response to 5 hydroxytryptamine (5-HT) and determined alterations in this response following stress. 5-HT stimulated rat colonic short-circuit current in a concentration dependent fashion (pD2 = 5.19). This response was subject to desensitization and was mimicked by the indolealkylamines with a rank order potency of 5-HT approximately alpha-methyl-5-HT > 5-carboxytryptamine approximately 5 methoxytryptamine. 2-Methyl-5-HT was a partial agonist. The colonic response to 5 HT was unaltered by methysergide (10 microM), ritanserin (0.1 microM), ondansetron (1 microM) or clozapine (10 microM), but was antagonized by the 5-HT4 receptor antagonists SB204070 (pD'2 = 9.32), GR113808 (pKb = 8.56), DAU6285 (pKb = 6.07) and SDZ205557 (pKb = 6.80). The response of colonic epithelial and oesophageal tunica muscularis mucosae to 5-HT is therefore mediated by a similar 5-HT4 receptor. Following wrap restraint stress, the colonic response to 5-HT became bimodal. Half of the preparations were hyper-responsive, while the rest were hypo-responsive to 5-HT. This 5-HT4 receptor may therefore be involved in stress related changes in fluid transport. PMID- 9726660 TI - Modulation of endothelial prostaglandin synthesis by corticotropin releasing factor and antagonists. AB - Corticotropin releasing factor (CRF) is a hypothalamic hormone that also displays autocrine/paracrine roles at peripheral sites. High concentrations of CRF have been identified in endothelial cells and other inflammatory tissues. We investigated the effects of CRF and antagonists in the regulation of prostaglandin synthesis in bovine aortic endothelial cells, and also characterized the binding of CRF in these cells. Interleukin-1alpha increased prostacyclin (prostaglandin I2) synthesis in endothelial cells and this response to interleukin-1alpha was abolished by simultaneous exposure to CRF. The effect of CRF on interleukin-1alpha-induced prostaglandin synthesis was antagonised by the CRF receptor antagonist alpha-helical CRF-(9-41). In addition, this as well as another CRF receptor antagonist, namely [D-Phe12]CRF-(12-41), when applied alone at low concentrations inhibited the interleukin-1alpha-induced prostaglandin synthesis similarly to CRF, suggesting partial agonistic action. Binding of [125I]-labeled CRF in endothelial cells was saturable and fitted a two sites model. Kd for the higher-affinity class of receptors was 0.2 +/- 0.02 nM, and Bmax 0.79 +/- 0.095 fmol/mg protein. The lower-affinity class of receptors had a Kd of 1.77 +/- 0.14 microM and Bmax 0.97 +/- 0.12 fmol/mg protein. These findings suggest a direct role for CRF in the local regulation of inflammation. PMID- 9726661 TI - Decreased protein kinase C activation mediates inhibitory effect of norathyriol on serotonin-mediated endothelial permeability. AB - We examined the mechanisms of norathyriol on the serotonin-induced increased permeability of rat heart endothelial cell monolayers. The present study showed that the activation of rat heart endothelial cell protein kinase C by phorbol myristate acetate led to the dose-dependent increase in endothelial permeability to albumin, an effect that was inhibited by staurosporine (a protein kinase inhibitor). Staurosporine also attenuated the serotonin-induced increase in permeability. Norathyriol abolished both serotonin- and phorbol myristate acetate induced permeability. We investigated whether norathyriol, by inhibiting protein kinase C activation, attenuated the serotonin-induced permeability. Immunofluorescence studies demonstrated that norathyriol prevented the redistribution of protein kinase C isozymes following stimulation with serotonin. Western blot analysis showed that norathyriol significantly inhibited the serotonin-induced translocation of the alpha protein kinase C isozyme from the cytosolic to the particulate fraction. In conclusion, norathyriol attenuates the serotonin-induced permeability of rat heart endothelial cells to macromolecules in association with inhibition of protein kinase C activation. This decrease in endothelial cell permeability may be one of the mechanisms for the protective effects of norathyriol against edema formation in response to inflammatory agonists in vivo. PMID- 9726662 TI - Highly selective sigma receptor ligands elevate inositol 1,4,5-trisphosphate production in rat cardiac myocytes. AB - Exposure of cardiac myocytes from adult rat ventricles to the highly selective, high affinity sigma receptor ligands 1S,2 R-cis-N-[2-(3,4-dichlorophenyl)ethyl]-N methyl-2-(1-pyrrolidinyl)-cycloh exylamine (BD-737) (0.1-100 nM) and N-[2-(3,4 dichlorophenyl)ethyl]-N,N',N'-trimethylethylenediamine (BD-1047) (0.01-10 nM), caused potentiation of electrically-evoked amplitudes of contraction and Ca2+ transients, while exposure to 100 nM BD-1047 caused attenuation of these amplitudes. In addition, BD-737 (1-100 nM) and BD-1047 (10-100 nM) caused an increase in the incidence of spontaneous twitches. These effects were inhibited when the incubation with BD-737 was done in the presence of the phospholipase C inhibitor, neomycin, or after pre-incubation with thapsigargin or caffeine which deplete the sarcoplasmic reticulum Ca2+ stores. Inositol 1,4,5-trisphosphate (IP3) production in cardiac myocytes was determined by the IP3 binding protein assay. Both substances caused an increase in the intracellular concentration of IP3. BD-737 caused a rapid transient increase to 3.2-fold in 1 min and stabilization at 2.1-fold of control thereafter. BD-1047 caused a gradual increase reaching 4.4-fold after 5 min. The results suggest that the effects of these sigma receptor ligands on contractility and spontaneous contractions are mediated by activation of phospholipase C and elevation of intracellular IP3 level. PMID- 9726663 TI - Inhibition of peroxynitrite-mediated tyrosine nitration by a novel pyrrolopyrimidine antioxidant. AB - Peroxynitrite is a cytotoxic, free radical species that is formed by the combination of superoxide and nitric oxide. The goal of the present study was to examine the ability of a novel antioxidant, U-101033E, to prevent peroxynitrite mediated oxidative damage of red blood cell membrane proteins. Treatment of red blood cell membranes with peroxynitrite resulted in oxidative damage as evidenced by the presence of both membrane protein cross-linking and nitration of tyrosine residues. Membrane protein cross-linking was the result of oxidation of sulfhydryl groups and was completely blocked by the addition of dithiothreitol. Dithiothreitol also prevented peroxynitrite-mediated nitration of tyrosine red blood cell proteins. U-101033E prevented nitrotyrosine formation in peroxynitrite treated red blood cell membrane proteins in a concentration-dependent manner, with maximal protection observed at 100 microM U-101033E. However, at a similar concentration where U-101033E prevented tyrosine nitration, it had little or no effect on membrane protein cross-linking. Our results suggest that U-101033E may be intercepting a peroxynitrite-derived reactive nitrogen species that is capable of nitrating tyrosine residues. The ability of U-101033E to prevent tyrosine nitration by peroxynitrite represents a new role for this class of antioxidants and suggests that the pyrrolopyrimidines may be useful in the treatment of diseases where peroxynitrite-mediated injury is implicated. PMID- 9726664 TI - Effect of adrenomedullin on cAMP and cGMP levels in rat cardiac myocytes and nonmyocytes. AB - The purpose of the present study was to determine if cardiac myocytes and nonmyocytes secrete adrenomedullin, to investigate the effects of adrenomedullin on cAMP and cGMP levels in cardiac myocytes and nonmyocytes, to study the effect of calcitonin gene-related peptide (CGRP) receptor antagonist CGRP-(8-37) and adrenomedullin-specific receptor antagonist, adrenomedullin-(22-52) on response to adrenomedullin and CGRP. Neonatal (days 1-2) cardiac myocytes and nonmyocytes were prepared from the ventricle of Wistar rats. Not only cardiac myocytes, but also nonmyocytes secrete almost equal amounts of adrenomedullin into the media. Both adrenomedullin and CGRP increased the cAMP levels, not the cGMP levels, both in the myocytes and nonmyocytes. In myocytes, CGRP-(8-37), almost completely inhibited the adrenomedullin- and CGRP-induced cAMP formation. In nonmyocytes, CGRP-(8-37) completely inhibited the cAMP levels induced by adrenomedullin and CGRP. More profound antagonistic effect of CGRP-(8-37) on cAMP levels induced by adrenomedullin was observed in nonmyocytes than in myocytes. In contrast, antagonistic effect of adrenomedullin-(22-52) for adrenomedullin-stimulated cAMP formation was considerably less potent than CGRP-(8-37) both in myocytes and nonmyocytes. Adrenomedullin-(22-52) did not affect the cAMP formation induced by CGRP either in myocytes or nonmyocytes. These results suggest that myocytes and nonmyocytes secrete adrenomedullin and that adrenomedullin increases cAMP levels possibly via different receptors in myocytes and nonmyocytes. PMID- 9726665 TI - Validity of visual stabilization conditions used with computerized dynamic platform posturography. AB - Temporal and spatial relationships between head motion and tilt of a visual surround were evaluated in conditions 3 and 6 of the sensory organization test (SOT) to determine the validity of visual stabilization during computerized dynamic platform posturography (CDP). Seven healthy subjects participated (mean age 42.4 years, SD = 19 years, range 25-72 years). Sagittal head translation and head pitch were evaluated separately with respect to displacement of the visual surround. Overall, the mean correlation between sagittal head motion and surround tilt was r = 0.68 for C3 and r = 0.93 for C6. The average phase (aggregate findings supported with trial-by-trial analysis), however, showed that sagittal head displacement leads motion of the visual surround ( + 452 ms for C3 and + 250 ms for C6). The mean correlation between head pitch and surround tilt was r = 0.30 for C3 and r = -0.72 for C6. Head pitch also leads visual surround motion (+567 ms for C3 and +539 ms for C6), but with greater variability than sagittal head translation. The results showed that visual stabilization was not consistently achieved during SOT conditions 3 and 6. The fundamental assumption that posture behaves as an inverted pendulum, with control of equilibrium originating in the lower extremities, does not appear to be applicable to the voluntary corrections of sway required during the SOT. PMID- 9726666 TI - The management of horizontal-canal paroxysmal positional vertigo. AB - Horizontal-canal paroxysmal positional vertigo (HC-PPV) is a vestibular syndrome due to canalolithiasis of the horizontal canal. The more common posterior-canal paroxysmal positional vertigo has a well defined and effective therapy, while there have been few reports on physical therapy for HC-PPV, and these have been tried in relatively few patients. We report the results of two different types of treatment of HC-PPV in 92 patients. A group of 21 untreated patients acted as a control group. One method, known as forced prolonged position (FPP), proposes liberating the affected canal by gravitation, and involves having the patient lie on the healthy side for many hours. The other method (the barbecue rotation) is a liberatory manoeuvre which proposes to expel the otoconia from the canal by rotating the patient 270 degrees around the longitudinal axis of the body in rapid steps of 90 degrees. FPP was successful in more than 70% of our patients; the barbecue rotation had slightly less successful but more immediate results. Both methods enable otoconial debris to migrate into the posterior canal. We suggest treating all patients with the two methods in succession. PMID- 9726667 TI - Actual and perceived postural sway during balance specific and non-specific proprioceptive stimulation. AB - A group of patients with balance complaints (n = 16) was compared with a group of normal subjects (n = 17) by means of posturography, subjective assessments of balance, anxiety and unsteadiness when standing on a force platform with eyes closed. Postural instability was induced by vibratory stimulation of the calf muscles (20, 40, 60, 80 and 100 Hz). As a control condition, the arm (biceps) was stimulated at similar frequencies. In order to control for arousal, blood pressure and heart beat were assessed. Furthermore, questionnaire responses on psychological measures were collected. Results showed clear differences between the groups in terms of imbalance and self-reports. However, the 2 groups displayed similar increases of imbalance during calf stimulation and no increase during arm stimulation. Patients generally rated less increase of unsteadiness when the calf was stimulated than did the controls. No differences in arousal were found between the groups or within conditions. Results are discussed in terms of the proposed desynchrony between symptoms and complaints. PMID- 9726668 TI - Postural reactions induced by vertical motion of visual scenes and the effects of weightlessness. AB - Postural reactions induced by vertical optokinetic stimulation were recorded for 5 subjects in a ground-based study, and for one astronaut before, during, and after a 25-day spaceflight. On the ground, the amplitude of visually-induced postural reactions generally increased with stimulus velocity and saturated around 60 degrees/s, with an angle of body tilt which never exceeded 2-3 degrees. For velocities higher than 20 degrees/s, backward body tilt during upgoing optokinetic stimulation was larger than forward body tilt during downgoing stimulation. In weightlessness, the angle of body tilt was reduced compared to ground values, but after the flight the postural reactions were larger than before the flight. If the limited angle of body tilt on Earth is due to an inhibition from the graviceptive inputs which do not confirm the visual inputs, the larger angle of tilt might reflect that this inhibition was less effective after spaceflight. This ineffectiveness might reflect a confusion between body tilt and translation as the result of adaptation to weightlessness. PMID- 9726669 TI - Unilateral vestibulotoxicity due to systemic gentamicin therapy. AB - Systemic gentamicin can cause acute bilateral, simultaneous, symmetrical loss of vestibular function manifested by symptoms and signs of chronic vestibular insufficiency (ataxia and oscillopsia). We report 6 patients presenting with ataxia and oscillopsia, but without a history of vertigo, who had severe unilateral loss of vestibular function on caloric testing. The absence of vertigo in these patients could be explained by two possible mechanisms: either, the unilateral loss of vestibular function was subacute, occurring over several days so that compensation could occur, or bilateral vestibular loss occurred which was then followed by asymmetrical recovery of vestibular function. The second hypothesis is supported by the observation that vestibular hair cells can regenerate after aminoglycoside damage. PMID- 9726670 TI - Subjective visual horizontal during follow-up after unilateral vestibular deafferentation with gentamicin. AB - The subjective visual horizontal (SVH) was measured by means of a small, rotatable, luminous line in darkness in the upright head and body position and at 10, 20 and 30 degrees of tilt to the right and left before, and repeatedly during a follow-up period of 1 year after intratympanic gentamicin instillations in 12 patients with recurrent vertigo attacks. This treatment caused a loss of the bithermal caloric responses on the diseased side. Shortly after treatment there was a significant tilt of SVH towards the treated side (group mean = 10.6 degrees). Repeated testing made it possible to characterize mathematically the changes with time for SVH. For the group of patients as a whole this otolithic component of vestibular compensation was best described by a power function, SVH = 8.65t(-0.16) degrees, where t is time in days after maximum tilt of SVH. After 1 year, SVH was still significantly tilted towards the treated side (group mean = 3.16 degrees). Gentamicin treatment also caused a significant reduction in the perception of head and body tilt towards the deafferented side, while the perception of tilt towards the healthy side did not show any significant changes. During follow-up there was a gradual improvement in the perception of tilt towards the treated side. However, a significant asymmetry in roll-tilt perception was still present 1 year after deafferentation. There was no correlation between SVH in the upright position and roll-tilt perception, suggesting that these parameters are to some extent dependent on different afferent input from the vestibular organ. They were also found to be complementary for the detection of vestibular disturbance. PMID- 9726671 TI - Antiviral treatment of idiopathic sudden sensorineural hearing loss: a prospective, randomized, double-blind clinical trial. AB - A subclinical viral labyrinthitis has been postulated in the literature to elicit Idiopathic Sudden Sensorineural Hearing Loss. An etiological role for the herpes virus family is assumed. Corticosteroids possess a limited beneficial effect on hearing recovery in ISSHL. In this study, the therapeutic value of the antiherpetic drug aciclovir (Zovirax) on hearing recovery in 44 ISSHL patients receiving prednisolone is evaluated in a multicentre clinical trial. The study is designed prospectively, randomized, double-blind and placebo-controlled. Subjective parameters include hearing recovery, a pressure sensation on the affected ear, disequilibrium or vertigo and tinnitus. Audiometric parameters include pure tone and speech audiometry. A one-year follow up is obtained. Both the pressure sensation and disequilibrium or vertigo have a good prognosis, but tinnitus, occurring in most patients, has a poor prognosis. Hearing recovery prognosis depends on the severity of initial hearing loss, and not on vestibular involvement. No beneficial effect from combining aciclovir with prednisolone can be established in ISSHL. PMID- 9726672 TI - Diagnostic potential of distortion product otoacoustic emissions in severe or profound sensorineural hearing loss. AB - To evaluate the potential of distortion product otoacoustic emissions (DPOAEs) in differential diagnosis of hearing loss, these were routinely measured in 232 ears of severe or profound sensorineural hearing loss. Normally recordable DPOAEs were found in 16 ears (8 patients) and the results were confirmed through retests after intervals; positive responses of transiently evoked otoacoustic emissions (TEOAEs) were additionally tested. The findings suggest that nerve deafness and hair cell deafness may be partly distinguishable. PMID- 9726673 TI - Sensitivity of auditory brainstem response in acoustic neuroma screening. AB - Auditory brainstem response (ABR) is the reference screening technique for acoustic neuromas, but because of a few false negatives and the increasing performance of magnetic resonance imaging (MRI), its role as the standard method has been questioned. We assessed sensitivity of screening tests in 89 patients with surgically proven acoustic neuromas. Sensitivity of ABR was 92%; 94% for extracanalicular neuromas and 77% for intracanalicular neuromas. For stapedius reflex (SR), sensitivity was 84% and for caloric vestibular response (CVR) 86%. The combined sensitivity of ABR + SR was 97% and of ABR + RS + CVR 98%. For false negatives, the greatest diameter including the intracanalicular portion was always less than 18 mm, with a mean of 15 mm, and none of these tumours reached the brainstem. For patients with unilateral cochleo-vestibular deficit, we propose ABR and SR as first-line screening tests. These tests are repeated at 6 months and at 1 year in the case of normal results. MRI is ordered for patients whose auditory threshold is too low and for those whose ABR or SR results favour retrocochlear disease. PMID- 9726674 TI - Histologic demonstration of glycosaminoglycans in inner ear fluids. AB - Hyaluronic acid is a glycosaminoglycan (GAG) which has been demonstrated by biochemical and histochemical methods to be present in inner ear fluids and tissues. However, consistent labeling of hyaluronic acid or other GAGs in inner ear fluid compartments in histologic sections has not been previously reported. Staining and characterization of GAGs in the normal inner ear fluids of humans and guinea pigs are described in this report. It was initially observed that alcian blue produces dense staining of the endolymphatic and perilymphatic fluid compartments in celloidin embedded material. Results obtained with alcian blue were subsequently compared to staining obtained with hematoxylin/eosin and periodic acid/Schiff base. Enzyme digestion was also performed with testicular hyaluronidase (bovine and ovine), streptomyces hyaluronidase, chondroitase ABC, and alpha-amylase. Results of the differential staining and enzyme digestion studies suggest that the substances in inner ear fluids that stain with alcian blue are a combination of chondroitin sulfate-4, -5 and/or -6 and hyaluronic acid. PMID- 9726675 TI - Acute effects of semicircular canal destruction on the cochlea, with and without preceding Pseudomonas aeruginosa exotoxin A treatment. AB - Acute electrophysiological and morphological changes in the cochlea following destruction of the semicircular canals (SCCs) were investigated to elucidate differences in cochlear vulnerability to surgical procedure under two conditions: normal healthy condition and a pathological condition induced by Pseudomonas aeruginosa exotoxin A (PaExoA). Frequency-specific auditory brainstem response recording and examination by light microscopy (LM) and scanning electron microscopy (SEM) were performed in the acute stage, immediately following, and 2, 5, and 10 days after the intervention. All normal healthy rats showed noticeable hearing loss immediately after the operation, predominantly in lower frequency areas, followed by complete recovery within 5 days. LM revealed rupture and slight distension of Reissner's membrane. SEM revealed considerable disarray of the stereocilia, especially on the outer hair cells (OHCs) of the third row in the upper half-turns of the cochlea. By contrast, all rats under pathological conditions showed delayed and incomplete hearing recovery from postoperative hearing loss. LM revealed various kinds of cochlear reaction, such as distension of Reissner's membrane, infiltration of inflammatory cells into the cochlea, and severe inflammatory change. Damage both of inner hair cells in the basal turn and of OHCs in the upper half-turns was more noticeable, compared with that under normal conditions. These findings indicate that SCC destruction under pathological conditions induced by PaExoA is detrimental to postoperative hearing recovery, even if the preoperative hearing level had appeared electrophysiologically normal. PMID- 9726676 TI - Effects of betahistine on vestibular receptors of the frog. AB - Betahistine is widely used in the symptomatic treatment of peripheral and central vestibular disorders. However, its remains unknown whether the drug can act directly on inner ear sensory organs. To this end, the effects of betahistine (10(-7)-10(-2) M) were examined on isolated preparations of frog semicircular canal mounted in a double-celled bath which allowed drug administration both in the endolymphatic and in the perilymphatic fluid. The effects of betahistine were evaluated by recording ampullar receptor potentials and nerve firing rate both at rest and during mechanical stimulation of the isolated preparation. The results demonstrated that endolymphatic administration of betahistine had no effect, whereas its perilymphatic administration could reduce greatly ampullar receptor resting discharge but had little effect on mechanically evoked responses. This observation may explain the anti-vertigo effects of betahistine. Vertigo is normally due to uncontrolled changes in vestibular receptor resting discharge. It is therefore probable that any factor able to reduce the resting firing rate of vestibular receptors and, in consequence, its variations, may have an anti vertigo action. PMID- 9726677 TI - Chronic study on the neuronal excitability of the cochlear nuclei of the cat following electrical stimulation. AB - To examine the safety of auditory brainstem prosthesis, the cat cochlear nuclei were implanted and stimulated chronically with bipolar surface electrodes using charge-balanced biphasic current pulses at rates of 250 pulses/s. The stimulation was continuous 16 h/day for up to 12 weeks. The electrically-evoked auditory brainstem response (EABR) was used to monitor neuronal excitability of the cochlear nuclei following the chronic electrical stimulation. The body weight, respiration, and body temperature of the cats were monitored throughout the experiment. The amplitudes and latencies of the EABR waves were measured fortnightly and compared before, during and after the electrical stimulation. The results showed that the respiration, body weight and body temperature of the cats remained within normal limits during the chronic stimulation. During the stimulation, no change was found in the EABR waveform, but a decreased threshold and wider dynamic range were observed after the stimulation. There was no significant change in the amplitudes and latencies of the EABR waves after stimulation. The present findings suggest that chronic bipolar electrical stimulation with surface electrodes at rates of 250 pulses/s is safe for neuronal excitability of the cochlear nuclei in the cat. PMID- 9726678 TI - Dose-dependent response of vestibular hair cells of guinea pigs following streptomycin ototoxiation. AB - Although the involvement of apoptosis has been suggested in the loss of vestibular hair cells due to aminoglycosides, dose-dependent effects of aminoglycosides have not been determined. We therefore examined dose-dependent effects of streptomycin on the degeneration of hair cells of guinea pig ampullar cristae using TUNEL stain and Hoechst nuclear stain. Streptomycin induced apoptosis of hair cells in a dose-dependent manner. Even following high-dose applications, most of the affected cells showed apoptotic features. Apoptosis may therefore play a predominant role in the deletion of vestibular hair cells affected by aminoglycosides. PMID- 9726679 TI - Morphometry of fish inner ear otoliths after development at 3g hypergravity. AB - Size and asymmetry (size difference between the left and right sides) of inner ear otoliths of larval cichlid fish were determined after a long-term stay in moderate hypergravity conditions (3g; centrifuge), in the course of which the animals completed their ontogenetic development from hatch to freely swimming. Neither the normal morphogenetic development nor the timely onset and gain of performance of swimming behaviour were impaired by the experimental conditions. However, both utricular and saccular otoliths (lapilli and sagittae, respectively) were significantly smaller after hyper-g exposure compared to 1g control specimens raised in parallel. The asymmetry of sagittae was significantly increased in the experimental animals, whereas the respective asymmetry of lapilli was pronouncedly decreased compared with the 1g controls. These findings suggest that growth and development of bilateral asymmetry of otoliths are guided by the environmental gravity vector. Some of the hyper-g animals revealed a kinetotic behaviour on transfer to normal 1g earth conditions, which was similar to the behaviour observed in previous experiments on the transfer from 1g to microgravity (parabolic aircraft flights). The lapillar asymmetry of kinetotic samples was found to be significantly higher than that of normally behaving experimental specimens. No differences in asymmetry of sagittae were obtained between the two groups. This supports an earlier theoretical concept, according to which human static space sickness might be based on asymmetric utricular otoliths. PMID- 9726680 TI - Production of otoconia in the endolymphatic sac in the Japanese red-bellied newt, Cynops pyrrhogaster: light and transmission electron microscopic study. AB - The formation of otoconia in the endolymphatic sac (ES) of the larval newt, Cynops pyrrhogaster, has been studied by light and transmission electron microscopy. Some of the epithelial cells of the ES contain an abundance of swollen vesicles, Golgi complexes, rough endoplasmic reticula and ribosomes at the late larval stages 50 and 51, approximately 26-30 days after eggs are laid. Five days later, at stage 52, crystals are present in the vacuoles between the epithelial cells. Serial sections indicate that these vacuoles actually form small canals which lie in the wall and join the lumen of the ES. Reconstruction of the ES shows that several canals are contained in the ES wall. At stage 56, about 72 days after eggs are laid, a large number of otoconia are present in the ES lumen, while the otoconia disappear from the canals. It appears that the otoconia are first produced in the canals and then released to the lumen. Some epithelial cells of the ES are thought to expel the organic and inorganic material to the canals to form the otoconia in situ. The process of formation of the otoconia in the ES is discussed. PMID- 9726681 TI - Presenilin-1 mRNA and beta-amyloid precursor protein mRNA are expressed in the developing rat olfactory and vestibulocochlear systems. AB - The mRNA expression of presenilin-1 (PS1) and beta-amyloid precursor protein (betaAPP) was investigated in the embryonic day 20 rat olfactory bulb, nasal cavity, and inner ear using in situ hybridization histochemistry. In the olfactory bulb, PS1 mRNA was strongly expressed in both olfactory bulb neuroepithelium and the differentiating olfactory bulb. In contrast, betaAPP mRNA was preferentially expressed in differentiating fields. In the nasal cavity, PS1 mRNA was strongly expressed throughout the olfactory epithelium, while betaAPP mRNA expression was concentrated in the middle part of the epithelium. In the membrane labyrinth of the inner ear, although PS1 mRNA was evenly distributed in both sensory epithelium and supporting cells, betaAPP mRNA was exclusively expressed in the sensory epithelium. These data suggest that PS1 is expressed earlier than betaAPP, and that PS1 and betaAPP co-operatively play pivotal roles in the development of the olfactory and vestibulocochlear systems. PMID- 9726682 TI - Prevention of vestibular deafferentation-induced spontaneous nystagmus with pretreatment of Ca2+ channel/N-methyl-D-aspartic acid receptor antagonists in guinea pigs. AB - Involvement of Ca2+ channel and N-methyl-D-aspartic acid (NMDA) receptors with induction of the spontaneous nystagmus (SN) after unilateral labyrinthectomy (UL) was evaluated by examining the effect of verapamil and MK-801 in guinea pigs. An injection of verapamil or MK-801 before the intratympanic application of arsanilate significantly (p < 0.05) suppressed the frequency of the SN towards the arsanilate-applied side. The frequency of the SN towards the intact side in these drug-treated animals was much lower than in the control animals until 36 h after the application of arsanilate. In addition, 60 days subsequent to induction of the UL, application of the drug before we injected the arsanilate into the opposite middle ear suppressed the SN towards the second arsanilate-injected side, but not towards the first injection side. We suggest that Ca2+ channel and NMDA receptor may be involved in the induction of the SN, and that the pretreatment of their antagonists could be applied in preventing the vestibular deafferentation-induced SN. PMID- 9726683 TI - Epidemiology and treatment of otitis media with effusion in children in the first year of primary school. AB - In this multicentre study we evaluated the prevalence and risk factors of otitis media with effusion (OME) in Italian school-children and the effectiveness of medical treatment of chronic OME with a new cephalosporin, ceftibuten. During two winter periods, 3413 children, aged 5 to 7 years, were examined for the presence of OME by means of pneumotoscopy and a portable, hand-held tympanometer. The prevalence of asymptomatic OME was 14.2%, with no difference as regards sex, age, month of examination or geographic area. Younger children had significantly more bilateral than unilateral effusion. A recent episode of acute otitis media and previous tonsillectomy or adenoidectomy were associated with an increased risk of OME in multivariate logistic regression models. The presence of OME was unrelated to such factors as birthweight, prematurity, sibling or parental history of allergy, duration of daycare attendance, family history of ear infections. After 12 weeks, 26.6% of children with OME still had middle-ear fluid: 52 were randomized to ceftibuten (9 mg/kg q.d. for 14 days) and 59 to no treatment (nasal saline drops allowed). Children treated with ceftibuten had a significantly better resolution of middle-ear effusion after 4 and 8 weeks. As mass screening programmes for OME in the year of school entry are questioned, a focus only on children with known risk factors seems advisable. Ceftibuten can be useful in reducing the duration of middle-ear effusion. PMID- 9726684 TI - Gammadelta T cell receptor repertoire in middle ear effusions in children. AB - In order to elucidate the immune response in otitis media with effusion, polymerase chain reaction was employed to examine gammadelta T cell receptor repertoire in the middle ear effusions of patients with otitis media with effusion. RNAs were extracted from 13 middle ear effusions of 10 children with otitis media with effusion. Vgamma2 was the most frequently used Vgamma gene. As for Vdelta gene usage, Vdelta2 amplification gave the strongest signal in 10 out of 13 samples. The results suggest that gammadelta T cells bearing Vgamma2/Vdelta2 T cell receptors accumulate in the middle ear effusions in children, and that these T cells may respond to certain bacteria or bacterial products in the middle ear. PMID- 9726685 TI - Exchange rates of gases across the tympanic membrane in rhesus monkeys. AB - The exchange rates of CO2 and He across the tympanic membrane were estimated in 5 monkeys. For these experiments, the monkey was anesthetized and one arm of a polyethylene "T" tube was introduced into the external canal of the test ear and sealed to the ambient environment with wax. One arm of the T tube was attached to a pressure transducer and the other to an argon gas source via a valve. Silica tubing sealed within the probe provided periodic gas samples for composition analysis by an online mass spectrometer. Prior to each experiment, the probe was washed with Argon. In 5 experiments the probe was sealed within the external canal of animals with physiological middle ear gas compositions, and in 5 experiments the probe was sealed within the external canal of animals whose middle ears were partially washed with He. The gas in the probe was sampled and analyzed at 10-min intervals for up to 4 h. The results documented a significant increase in the percentage composition of CO2 but not He in the experiments conducted with physiological middle ear gas compositions, and increases in both He and CO2 in the experiments conducted after the middle ear was washed with He. Estimated, average exchange constants for He and CO2 were 0.0005 microl/min/mmHg and 0.0103 microl/min/mmHg, respectively. Using data from previous experiments, the relative trans-mucosal to trans-tympanic membrane gas exchange rates in monkeys are estimated to be in the order of 10:1 for inert gases and 180:1 for chemically active gases. PMID- 9726686 TI - Ventilator-associated sinusitis: antroscopic findings and bacteriology when excluding contaminants. AB - The objective of this study was to investigate maxillary sinus infectious or inflammatory disease in long-term mechanically ventilated patients, using a prospective case series design. The subjects were 33 critically ill patients receiving ventilator treatment for more than 7 days, in the general and neurosurgical intensive care units of a tertiary care hospital. Bilateral antroscopies were performed, and antral secretion and mucosal samples were collected for bacterial culturing. Different stages of inflammatory disease were found in 85% of the antra. Infectious sinusitis was diagnosed in 6%, with a predominance of mixed anaerobic infections. Monoisolates of anaerobic or facultative anaerobic bacteria were found in 5% of the antra without clinical signs of infection. In conclusion, more stringent requirements are needed for the diagnosis of infectious sinusitis in ventilator-treated patients. With improved diagnostic techniques there was a significantly lower frequency of infectious sinusitis than presented in previous reports. The bacteria predominating as infectious agents were different to those in previous reports. Furthermore, reactive inflammatory disease with bacterial colonization occurred. PMID- 9726687 TI - Children with nocturnal upper airway obstruction: postoperative orthodontic and respiratory improvement. AB - Twenty children, aged 4-9 years, underwent adeno/tonsillectomy because of unequivocal anamnestic nocturnal obstructive breathing. Preoperatively, apnea hypopnea index was > 5 in 10 cases only, AI > 1 in 17. Nineteen children had signs of increased respiratory labour in movement recordings and inspiratory EMG activity. Oxygen desaturation index was 0 in 7 children, and nadir SaO2 was > or = 90% in 10. Cephalometry and dentition study models initially revealed significant changes, chiefly lateral cross-bite (n = 11) and vertical growth direction of the mandible. Tonsillar size or duration of disease was not correlated with the severity of polysomnographic findings, nor were orthodontic variables. Symptoms disappeared promptly postoperatively. After one year, respiratory recordings were normalized or improved in the majority of children, and orthodontic variables normalized or improved in all children. CONCLUSION: Oximetry and airflow recordings may be normal in children who benefit from treatment of anamnestic nocturnal obstruction. Craniofacial deformities are common and improve significantly with surgical treatment of the airway obstruction. PMID- 9726688 TI - Membranous basal cell adenoma of the salivary gland: a clinicopathologic study of 12 cases. AB - Twelve cases with membranous basal cell adenoma of the salivary gland were analysed clinicopathologically. Its histology was characterized by palisading of peripheral cells and an excessive hyaline basal membrane. The differential diagnosis involved the solid variant of basal cell adenoma, basal cell adenocarcinoma, solid subtype of adenoid cystic carcinoma and basaloid squamous cell carcinoma. Four cases had coexisting dermal cylindromas of the scalp. Screening for skin lesions of the scalp or other locations is suggested. In 8 cases, the tumors demonstrated multifocal origin. Total parotidectomy rather than superficial parotidectomy is suggested to avoid the recurrence of the tumor. One case showed malignant transformation and cervical lymph node metastases. A close follow-up after treatment is necessary. PMID- 9726689 TI - Comparison and evolution of perceptual and acoustic characteristics of voice after supracricoid partial laryngectomy with cricohyoidoepiglottopexy. AB - The present prospective study, based on a series of 12 male patients managed with supracricoid partial laryngectomy with cricohyoidoepiglottopexy (SCPL-CHEP), was designed i) to compare the perceptual and acoustic parameters before surgery and at 6 months after sugery, ii) to evaluate the evolution of the perceptual and acoustic parameters between 6 and 18 months postoperatively, and iii) to determine the correlations between the perceptual and acoustic parameters preoperatively and at 18 months postoperatively. The roughness was found to be significantly worsened after SCPL-CHEP. The jitter, shimmer, noise-to-harmonic ratio, and degree of voiceless increased significantly after SCPL-CHEP. Neither acoustic nor perceptual parameters varied significantly between 6 and 18 months postoperatively. Preoperatively, a strong statistical correlation was found between grade, roughness and strain and all acoustic parameters but F0. Breathiness was statistically correlated with all acoustic parameters but jitter. Postoperatively the only statistical correlation noted was between roughness and F0. PMID- 9726690 TI - Cytotoxic and genotoxic effects of paclitaxel (Taxol) and radiation in a squamous cell carcinoma cell line of the larynx. AB - Paclitaxel (Taxol) is an antimicrotubular agent which blocks the cells in the G2/M phase of the cell cycle. Because of this mechanism it is presumed that this drug could function as a radiation sensitizer. The cytotoxic and genotoxic effects of paclitaxel and a combination of paclitaxel and radiation were studied in the human laryngeal carcinoma cell line HLac 79. The growth of the cells was significantly reduced at concentrations of paclitaxel as low as 10 nM. Flow cytometry data showed a G2/M block after exposure to paclitaxel. Radiation at 12 and 24 h after drug treatment exerted an additive but no radiation sensitizing effect. As genotoxic effect paclitaxel induced multinucleated cells, possibly in a synergistic manner, at low concentrations (10 nM) and radiation doses up to 3 Gy. PMID- 9726691 TI - Caroverine in tinnitus treatment. PMID- 9726692 TI - Lack of drug-drug interaction between cerivastatin and nifedipine. AB - Cerivastatin is a novel, potent HMG-CoA reductase inhibitor. It is primarily cleared via demethylation and hydroxylation with involvement of cytochrome P450 (CYP) 3A4 and subsequent biliary and renal excretion of the metabolites. Both cerivastatin and the dihydropyridine calcium antagonist nifedipine, which is primarily metabolized by CYP 3A4, are used concomitantly in the prevention and therapy of coronary heart disease. To study the drug-drug interaction potential, the mutual effects of cerivastatin and nifedipine were investigated in a controlled, randomized, non-blind 3-way crossover study in healthy male subjects. Single oral doses of 0.3 mg cerivastatin or of 60 mg nifedipine were administered either alone or concomitantly under fasting conditions. The mean AUC- and Cmax ratios (combination treatment versus monotherapy) including 90% confidence intervals were 1.04 (0.98 - 1.10) and 1.00 (0.93 - 1.07) for cerivastatin, and 0.98 (0.73 - 1.32) and 0.95 (0.80 - 1. 13) for nifedipine, respectively. Our results indicate that no mutual drug-drug interaction between cerivastatin and nifedipine occurs. PMID- 9726693 TI - Pharmacokinetics of dexibuprofen administered as 200 mg and 400 mg film-coated tablets in healthy volunteers. AB - The pharmacokinetic properties of 2 film-coated preparations containing 200 mg and 400 mg dexibuprofen were compared in a single-dose, crossover study in 16 healthy, male volunteers. Dexibuprofen was absorbed rapidly (tmax 2.1 - 2.2 hours) reaching maximum concentrations of 12.4 microg/ml (200 mg), respectively 12.0 microg/ml (400 mg dose adjusted). For the characteristics AUC(0-12h) and AUC(0-infinity) arithmetic means of 49.2 (microg) x (h/ml)(200 mg) and 48.2 (microg) x (h/ml)(400 mg dose-adjusted), respectively 50.5 (microg) x (h/ml)(200 mg), and 49.2 (microg) x (h/ml)(400 mg) were calculated. No relevant differences for the pharmacokinetic characteristics terminal half-life, clearance, volume of distribution, and mean residence time were observed. A linear dose-relationship was shown over the investigated dose range. Mean ratios after dosage adjustment of the test preparation using the "2 one-sided t-tests" procedure were calculated. Bioequivalence was assessed for AUC(0-12h) with a mean ratio of 97.7% (90% CI: 92.4 - 103.3%), for AUC(0-infinity) with 97.1% (90% CI: 91.4 - 103.1%), and for Cmax with 97.5% (90% CI: 91.7 - 103.8%). Both dexibuprofen preparations were well tolerated. No changes in hematological and biochemical parameters were detected. PMID- 9726694 TI - Transsynovial kinetics of lonazolac and its hydroxy metabolite in synovitis patients. AB - OBJECTIVE: The study was designed to characterize the synovial distribution profiles and kinetics of the non-steroidal antiinflammatory agent, lonazolac, in patients with synovitis after multiple dosing with 300 mg tablets of lonazolac calcium salt. METHODS: Forty patients (36 male, 4 female) aged 21 to 50 years (mean: 38+/-9 years) undergoing arthroscopy of the knee joint for surgical reasons were given 7 total doses of drug administered as 300 mg oral tablets of lonazolac-calcium taken twice daily. Patients were assigned to one of 4 treatment groups (n = 10) in which arthroscopy was carried out 1, 2, 6, or 12 h after the seventh lonazolac dose. Samples of blood, synovial fluid, and synovial membrane were obtained during each operation and used to determine total concentrations of lonazolac and its main metabolite in plasma and synovial fluid by HPLC assay with UV detection. Free lonazolac concentrations in body fluids were determined after ultrafiltration by the same HPLC technique using a fluorescence detector. Tissue concentrations were assayed after additional steps using solvent and solid phase extractions. Total protein contents in plasma and synovial fluid were measured spectrophotometrically. RESULTS: Plasma drug levels were highest at 1 hour after dosing with mean peak concentrations of 1.8 mg/l total lonazolac, 1.2 mg/l total metabolite, and 9 microg/l free lonazolac. Profiles indicated a biphasic decline. Concentration vs. time profiles in synovial fluid were flattened compared to plasma profiles with mean peak values of 440 microg/l total lonazolac, 370 microg/l total metabolite, and 7 microg/l free lonazolac attained 2 hours after dosing. The mean unbound fraction of lonazolac was higher in synovial fluid (1.9%) compared to plasma (0.7%). Transsynovial partition coefficients increased continuously during a dosing interval from 0.16 to 3.15 for total lonazolac and from 0.56 to 5.05 for free lonazolac. Mean total protein contents for each group of patients ranged from 70 to 76 g/l for plasma and 32 to 42 g/l for synovial fluid. Total drug concentrations in synovial membrane were highest in tissues obtained 1 hour after dosing with mean values of approximately 1.0 microg/g dry weight. Tissue samples obtained at later times indicated that lonazolac profiles in tissue more closely resemble profiles obtained for plasma than for synovial fluid. Protein concentration ratios (synovial fluid : plasma) were between 0.45 and 0.58. Except for the absorption phase, transsynovial drug partition coefficients were always higher than the protein concentration ratios. CONCLUSIONS: Protein content is not an important factor for drug partition into inflamed joints after multiple dosing with lonazolac. Lonazolac distributes well into synovial fluid with therapeutically effective concentrations of unbound drug measured within 2 hours after dosing. Total lonazolac levels in synovial fluid exceed those measured in plasma at 6 to 12 hours after administration. PMID- 9726695 TI - Blood pressure and aortic elastic properties--verapamil SR/trandolapril compared to a metoprolol/hydrochlorothiazide combination therapy. AB - The effects of 2 fixed antihypertensive combination drugs on blood pressure and aortic elastic properties were compared in 2 parallel groups. Twenty-six patients for 6 months received a calcium antagonist plus ACE inhibitor (verapamil SR 180 mg/trandolapril 1 mg (Vera/Tran)) and 25 patients a beta-adrenoceptor antagonist plus diuretic (metoprolol 100 mg/hydrochlorothiazide 12.5 mg (Meto/HCTZ)). In addition to blood pressure (SBP, DBP), carotidofemoral pulse wave velocity (PWV) was assessed non-invasively. Total peripheral resistance (TPR) was determined from cardiac output derived by electrical impedance cardiography. Sitting DBP decreased for -14.4 mmHg following Vera/Tran compared with -9.2 mmHg following Meto/HCTZ (p = 0.02 for difference between treatments). Blood pressure was normalized (i.e. DBP < 90 mmHg) in 69% of patients with Vera/Tran and in 52% with Meto/HCTZ. PWV was lowered with Vera/Tran to a higher extent than with Meto/HCTZ (differences between group means -0.46 to -0.98 m/sec, statistically not significant). Vera/Tran induced a decrease in TPR of about 15% of baseline values, whereas Meto/HCTZ showed no influence. Treatment-related adverse events following Meto/HCTZ were bradycardia and associated symptoms; following Vera/Tran these were cough and edema in 1 case each. In the Meto/HCTZ group, there were more withdrawals/drop-outs (9/25) than in the Vera/Tran group (2/26). The somewhat more intense reduction in PWV with Vera/Tran is indicative of an increase in aortic elastic properties associated with the more potent decrease in BP. In the present study, the combination of calcium antagonist plus ACE inhibitor was found to be an effective and well tolerated antihypertensive regimen and in these respects appears to have some advantages compared with a combination of beta-blocker plus diuretic. PMID- 9726696 TI - Pharmacokinetic characteristics of cisapride in elderly patients. AB - We demonstrated pharmacokinetic characteristics of cisapride in elderly patients (85+/-6 years) who had suffered from constipation based on intestinal motility dysfunction. After a single p.o. dose of 2.5 mg cisapride tablet on the first day, plasma unchanged cisapride concentrations were detected at 9 points within 24 hours, using high-performance liquid chromatography method. Subsequently, the patients were treated with cisapride 3 times daily during next 26 days (from the day 2 to the day 27). On the 28th day, a cisapride tablet was given once in the morning and blood samples were obtained with the same procedure as that on day 1. The obtained elimination half-life averaged 21.8+/-10.6 hours, which seems to be considerably longer than those previously reported. The area under the time concentration curve (AUC) on the day 28 was 2.7 times larger than AUC on the day 1, suggesting that plasma cisapride concentrations can become higher than expected by physicians when given 3 times daily to extremely elderly patients. Because cisapride sometimes induces fatal ventricular arrhythmias and sudden death in a concentration-dependent manner, this medicine should be administered once or twice per day to such patients. PMID- 9726698 TI - Beneficial effects of acarbose on familiar hypertriglyceridemias. AB - Elevated serum triglyceride levels may be related to the following clinical features: increased blood coagulation and viscosity, increased serum fibrinogen levels, decreased fibrinolysis, and for serum levels over 1000 mg/dl, a strong increase of acute pancreatitis rate. Pharmacological choice among the numerous drugs to treat hypertriglyceridemias is currently debated. Our study was aimed to assess the therapeutic efficacy of acarbose in the treatment of non-diabetic subjects, affected by familiar hypertriglyceridemia (FH). We studied 18 non diabetic patients (10 males, 8 females; mean age 57.61+/-6.85 years) without family history of diabetes mellitus affected by familiar hypertriglyceridemia. The study protocol planned a treatment period of 20 weeks, divided into five 4 week courses and made up as follows: diet plus acarbose therapy (4 weeks); diet therapy alone (4 weeks) alternatively. In the second and fourth 4-week courses diet plus acarbose were administered, while diet therapy alone was administered in the first, third, and fifth 4-week courses. Acarbose doses consisted of 50 mg (1/2 pill) twice daily. Mean serum triglyceride levels, after first month of dietary treatment, underwent a significant reduction from 481.5 +/- 67.1 mg/dl to 389.5 +/- 62.7 mg/dl, even if they did not reach the optimal levels to keep on the dietary therapy alone. After the first month of treatment with acarbose associated to diet, we observed a further reduction of serum triglycerides levels (p = 0.02). When diet alone was administered, mean triglyceride serum levels underwent a significant enhancement (p = 0.003). Restarting for the second time the association treatment, we observed a noteworthy reduction of mean serum triglyceride levels (p = 0.0001). Acarbose acts on the pathogenesis of FH, lowering the production of endogenous triglycerides. Our data suggested that acarbose can be considered a valid therapeutic tool in the treatment of familiar hypertriglyceridemias, also in non-diabetic patients. PMID- 9726697 TI - Unpredictable response to vasodilator therapy in primary pulmonary hypertension. AB - A 49-year-old man with severe primary pulmonary hypertension unresponsive to multiple vasodilators showed beneficial response to intravenous (i.v.) isoproterenol with significant decrease in the pulmonary vascular resistance and increase in the cardiac output. However, the patient became dependent on i.v. isoproterenol and eventually died suddenly following an episode of bradyarrhythmia. Autopsy confirmed severe primary pulmonary hypertension. As the response to vasodilator therapy can be unpredictable in PPH, sequential hemodynamic evaluation of drugs is necessary for finding an optimal therapeutic agent. PMID- 9726699 TI - The CYP3A4 inhibitor intraconazole does not affect the pharmacokinetics of a new calcium-sensitizing drug levosimendan. AB - Itraconazole is a potent inhibitor of CYP3A4 isoenzyme and it can cause clinically significant interactions with some other drugs. Levosimendan is a new calcium-sensitizing drug intended for congestive heart failure. We aimed to study possible interactions of itraconazole with levosimendan in healthy volunteers. Twelve healthy male volunteers were included into a randomized, double-blind, two phase crossover study. A wash-out period of 4 weeks was held between the phases. The subjects were given orally itraconazole 200 mg or placebo daily for 5 days. On the fifth day, they received a single oral dose of 2 mg of levosimendan. Levosimendan plasma concentrations were determined up to 12 hours and ECG, heart rate, and blood pressure followed-up to 8 hours after intake of levosimendan. Itraconazole had no significant effects on the pharmacokinetic parameters of levosimendan. Neither were there any differences in heart rate, PQ-, QTc- or QRS intervals between the placebo and itraconazole phases. The systolic blood pressure was decreased slightly more (p < 0.05) during the itraconazole phase than during the placebo phase. In conclusion, because the potent CYP3A4 inhibitor itraconazole had no significant pharmacokinetic interaction with levosimendan, interactions with CYP3A4 inhibitor, and oral levosimendan are unlikely. PMID- 9726700 TI - Asymmetry of the mean-variability tradeoff raises questions about the model in investigations of individual bioequivalence. AB - Tradeoff between changes of intraindividual variations of 2 drug formulations and of the difference between their means is a characteristic of a procedure suggested for the determination of individual bioequivalence [Schall and Luus 1993] and to be proposed by the Food and Drug Administration for adoption. Hauck et al. [1996] investigated properties of the tradeoff. Their procedure was applied and extended in the present study. The tradeoff was shown to be asymmetric. Notably, a small change in intrasubject variations can elicit, under various conditions, a comparatively large change in the allowable difference between means which can still be compatible with the declaration of bioequivalence. For instance, when the intraindividual coefficients of variations are 40% and 38% for the reference and test formulations, respectively, the allowable difference between means may increase, as a benefit, by 12.3%. A penalty by 11.2% is elicited if the intrasubject variations of the reference and test products are 40 and 42%, respectively. In addition, 4-period crossover trials were simulated. Ratios of estimated variances of the 2 formulations followed an F-distribution. Distributions of changes in allowable deviations between means were calculated from the tradeoff relationships; generally substantial changes were noted with high probabilities. For example, with an intraindividual variation of 30% there is an estimated 37% probability that a benefit of 10% increase, or larger, is gained by chance in the allowable difference between means, and an additional 36% probability that a penalty of a 10%, or larger, decrease in the allowable difference is suffered. With an intrasubject variation of 40%, the estimated probabilities are 42% and an additional 42% for a 10% expansion and contraction, respectively, of the allowable difference between means. Consequently, the strong asymmetry of the tradeoff could result in very large probabilities for benefits and penalties. Therefore, the investigated model assessing individual bioequivalence does not appear to be suitable for implementation. PMID- 9726701 TI - A new route, jet injection for anesthetic induction in children. IV. Midazolam plasma levels. AB - The jet injector route for midazolam was used in 40 children 1 - 6 years of age undergoing various short duration surgical procedures. A randomly selected dose of 100, 150, and 200 microg/kg was given by a jet injector and compared to 80 microg/kg by conventional i.m. injection with syringe and needle to induce sedation/anesthesia. Because of clinical limitations, plasma midazolam levels were measured for only up to 32 min post-injection. Peak levels were 70.4, 105.8, 157.3, and 135.5 ng/ml in the corresponding groups. Plasma levels reached their peak faster after 200 microg/kg jet injection than after 80microg/kg i.m. midazolam. Furthermore, midazolam plasma levels were sustained longer after 200 microg/kg by jet injection. Larger doses of midazolam are required by jet injection than by i.m. injection. Individual subjects showed considerable variability in plasma levels of midazolam by both methods of administration, although jet injection was more convenient and less traumatic. PMID- 9726702 TI - Middle ear gas--its composition in the normal and in the tubulated ear. A methodological and clinical study. PMID- 9726703 TI - Evidence for the needs of out-of-hospital thrombosis prophylaxis: introduction. PMID- 9726704 TI - The incidence of symptomatic venous thromboembolism after enoxaparin prophylaxis in lower extremity arthroplasty: a cohort study of 1,984 patients. Canadian Collaborative Group. PMID- 9726705 TI - Ultrasonographic screening for deep vein thrombosis following arthroplasty fails to reduce posthospital thromboembolic complications: the Postarthroplasty Screening Study (PASS). PMID- 9726706 TI - Out-of-hospital prognosis after hip surgery in patients with normal venography at discharge: clinical outcome at 3 months. PMID- 9726707 TI - Out-of-hospital prophylaxis with low-molecular-weight heparin in hip surgery: the French study--venographic outcome at 35 days. PMID- 9726708 TI - Out-of-hospital prophylaxis with low-molecular-weight heparin in hip surgery: the Swedish study. PMID- 9726709 TI - Busywork feeds upon itself. PMID- 9726710 TI - Cardiac effects of beta-agonists in patients with COPD. PMID- 9726711 TI - Identifying smokers at risk for developing airway obstruction. PMID- 9726712 TI - Duration of glucocorticoid treatment and outcome in sepsis: is the right drug used the wrong way? PMID- 9726713 TI - Suspended life or extending death? PMID- 9726714 TI - Timing tracheotomy: calendar watching or individualization of care? PMID- 9726715 TI - Can tuberculosis cause sarcoidosis? New techniques try to answer an old question. PMID- 9726716 TI - The effect of adding ipratropium bromide to salbutamol in the treatment of acute asthma: a pooled analysis of three trials. AB - OBJECTIVE: To assess the effect on FEV1 and clinical outcomes of adding ipratropium bromide to salbutamol in the treatment of acute asthma. METHODS: We conducted a pooled analysis of three randomized double-blinded clinical trials conducted in the United States, Canada, and New Zealand. The studies enrolled 1,064 patients aged 18 to 55 years who presented at the emergency department with acute asthma. Patients were randomized to treatment with a combination of nebulized 2.5 mg salbutamol plus 0.5 mg ipratropium bromide, or 2.5 mg salbutamol alone. Medications were administered at baseline and, in the US study, at 45 min. FEV1 was measured at baseline, 45 min, and 90 min. Patients were followed up for 48 h after hospital discharge for occurrence of asthma exacerbation and hospitalization. RESULTS: Treatment groups were comparable at baseline. Of the 1,064 patients randomized, 1,015 patients (95%) remained in the study for measurement at 45 min, and 961 patients (90%) completed the final measurement at 90 min. Comparison of overall improvement in FEV1 at 45 min indicated a better response for patients receiving combination therapy (mean difference=43 mL, 95% confidence interval [CI]=-20, 107). The distribution of change in FEV1 was skewed by a small number of patients with extreme values (38 of 1,064=3.6%) that may have been due to unreliable lung function testing. Removing these outliers produced a larger and more precise estimate of effect (mean difference=55 mL, 95% CI=2,107). Because the distribution was skewed, we performed nonparametric analyses that showed evidence of a beneficial effect of combination therapy. The difference between median values at 45 min is 40 mL (Wilcoxon p value=0.03). In addition, 4.9% (95% CI=-1%, 11%) more patients in the combination group achieved at least 20% of their potential improvement, as measured by the difference between their baseline FEV1 and their predicted FEV1. Patients receiving combination therapy had lower risk for each of three clinical outcomes: the need for additional treatment (relative risk [RR]=0.92, 95% CI=0.84, 1.0), risk of asthma exacerbation (RR=0.84, 95% CI=0.67, 1.04), and risk of hospitalization (RR=0.80, 95% CI=0.61, 1.06). CONCLUSION: Adding ipratropium bromide to salbutamol in the treatment of acute asthma produces a small improvement in lung function, and reduces the risk of the need for additional treatment, subsequent asthma exacerbations, and hospitalizations. These apparent benefits of adding ipratropium bromide were independent of the amount of beta-agonist that had been used earlier in the attack, and possibly related to a recent upper respiratory tract infection. Confirmatory studies are needed, especially for clinical outcomes. PMID- 9726717 TI - Influence of beclomethasone and salmeterol on the perception of methacholine induced bronchoconstriction. AB - BACKGROUND: Patient evaluation of asthma severity and medication needs is mostly based on respiratory symptoms and may be influenced by changes in perception of bronchoconstriction-induced sensations. However, the influence of asthma medication on the ability to perceive symptoms is still to be documented. This study evaluated the effects of short-term and regular use of salmeterol on the perception of methacholine-induced bronchoconstriction (MIB) in subjects with mild asthma, using inhaled salbutamol on an "as required" basis (n=15), and in subjects with moderate asthma, using daily inhaled beclomethasone (mean daily dose, 640 microg; n=15) in addition to salbutamol to control their asthma. METHODS: Methacholine challenges (MC) were performed at entry into the study, and then before, 1, and 12 h following inhalation of 50 microg of salmeterol or a placebo, after a 15-day baseline period; and after 4 weeks of twice daily use of those treatments. The measurements were then repeated with the alternate treatment after a 15-day washout period. Finally, a last MC was performed after another 15-day washout period. For each MC, the perception score of bronchoconstriction-associated breathlessness at 20% fall in FEV1 (PS20) was evaluated on a modified Borg scale from 0 to 10. RESULTS: Subjects using regular beclomethasone had a higher baseline PS20 than those using only salbutamol (means: 3.06 0.06 and 2.01+/-0.07, p=0.0001). Short- and long-term use of salmeterol did not change significantly the PS20 compared with placebo (p>0.05) in either group (with or without corticosteroid). Although there were some intraindividual variations, mean PS20 did not vary significantly throughout the study. CONCLUSION: These observations show that the perception of bronchoconstriction-associated breathlessness is not influenced by regular use of salmeterol. Patients using inhaled corticosteroids show a greater perception of MIB. PMID- 9726719 TI - Prevalence of physician-diagnosed asthma in Saskatchewan, 1981 to 1990. AB - STUDY OBJECTIVE: To examine trends in asthma prevalence in the Province of Saskatchewan using the Medical Claim Insurance Branch (MCIB) database. DESIGN: For each calendar year from 1981 to 1990, first visits to physicians for asthma were identified from the MCIB database. Age- and sex-specific prevalence rates were obtained for each calendar year by dividing the number of first asthma visits by the number of subjects in the age- and sex-specific group insured by the Provincial Government of Saskatchewan in that calendar year. RESULTS: Asthma prevalence increased in children and adults from 1981 to 1990. In the calendar year 1990, the prevalence of asthma was 5.1% in children < or = 4 years old, 4.4% in children 5 to 14 years old, 2.2% in young adults 15 to 34 years old, and 1.9% in adults 35 to 64 years old. Boys had higher asthma prevalence than girls in the age groups 0 to 4 years and 5 to 14 years, but it was reversed in older age groups, with women having greater asthma prevalence than men. Asthma diagnoses were verified by checking for asthma-related drug purchase in the Prescription Drug Plan database. Among the 0- to 4-year-old children with physician-diagnosed asthma, 57.3% purchased at least one asthma-related drug in 1981 and 72.3% purchased that in 1990. Among adults, asthma-related drug purchase was >73% in 1990. CONCLUSIONS: Asthma prevalence increased in children and adults in the province of Saskatchewan. Reasons for the increase are not clear and further studies are required to determine factors related to the increase. PMID- 9726718 TI - Hemodynamic, cardiac, and electrolyte effects of low-dose aerosolized terbutaline sulfate in asthmatic patients. AB - STUDY OBJECTIVE: Aerosolized beta2-agonists have been associated with increased morbidity in asthmatics. These drugs cause transient increases in heart rate and decreases in serum potassium levels after these drugs are first utilized. This study is designed to elucidate whether beta-adrenergic tolerance to the hemodynamic, cardiac, and electrolyte effects of inhaled terbutaline occurs during 14 days of maintenance therapy. DESIGN: Eight patients with stable asthma weaned off beta2-agonist therapy were studied in a randomized, double-blinded, placebo-controlled study utilizing aerosolized terbutaline, 400 microg q6h. Hemodynamic measurements and M-mode echocardiography were performed before and 15 and 30 min after the initial dose of terbutaline or placebo and after a dose of aerosolized terbutaline after 14 days of aerosolized terbutaline maintenance therapy. Holter monitors were worn on the first day of placebo or terbutaline therapy and on day 14 of terbutaline therapy. Plasma potassium, bicarbonate, and glucose levels, pH, renin activity, and serum insulin and aldosterone levels were measured before and after 24 and 48 h after terbutaline or placebo therapy and after 14 days of aerosolized terbutaline maintenance therapy. RESULTS: Terbutaline increased cardiac index and decreased systemic vascular resistance greater after 14 days of therapy compared with the first dose (5.2+/-0.5 vs 4.4+/ 0.6 L/min/m2; p<0.05; and 760+/-62 vs 1,016+/-118 dyne x s x cm(-5), p<0.01). After 14 days of terbutaline therapy, the mean maximum heart rate and number of episodes of heart rate > 100 beats/min were higher compared with the other study day (p<0.05). Plasma potassium level decreased from 4.29+/-0.09 to 3.65+/-0.16 mmol/L after 24 h of terbutaline and to 3.90+/-0.11 mmol/L after 48 h. Plasma potassium level returned to baseline after 14 d of terbutaline therapy. Plasma glucose and serum insulin levels rose significantly 24 h and 48 h after terbutaline and returned to baseline after 14 d of terbutaline therapy. Serum aldosterone level decreased significantly as serum potassium level decreased in the first 48 h of terbutaline therapy but returned to baseline levels after 14 d of terbutaline. CONCLUSIONS: Cardiovascular beta2-receptors in patients with stable asthma do not develop tolerance to the effects of low-dose aerosolized terbutaline after 14 days of maintenance therapy. In contrast, the homeostatic mechanisms regulating serum potassium develop tolerance to low-dose terbutaline maintenance therapy. Lack of cardiovascular tolerance to maintenance doses of aerosolized beta2-agonists may be important in increased morbidity if excessive amounts of these drugs are administered during asthma exacerbations. PMID- 9726720 TI - Childhood asthma and the indoor environment in a subtropical area. AB - STUDY OBJECTIVES: The objective of this study is to examine the relationship between indoor environmental factors and childhood asthma in a subtropical area. DESIGN: A case-control study was performed using participants of a prevalence survey that included 165 schoolchildren with asthma and 165 age- and gender matched control subjects. SETTING: The study was confined to 4,164 schoolchildren aged 6 to 12 years attending eight primary schools in Kaohsiung County rural municipalities who participated in a prevalence study concerning the health effects of the indoor environment. PARTICIPANTS: Cases (n=165) were defined as children with current asthma confirmed by a physician. Control subjects (n=165) were selected from the same school and class and matched for age and gender, and they did not have a previous diagnosis of asthma, history of physician-confirmed atopic diseases, persistent wheezing, cough, or phlegm, or reported chest illness, pneumonia, or bronchitis. MEASUREMENTS AND RESULTS: Information regarding the home environment was obtained using a structured written questionnaire, completed by the parents of the children. Of the many indoor environmental factors included in this study, only home dampness showed an association with asthma (odds ratio=2.65). CONCLUSIONS: We conclude that dampness in the home is a new public health issue in subtropical areas. PMID- 9726721 TI - Follow-up of occupational asthma after removal from or diminution of exposure to the responsible agent: relevance of the length of the interval from cessation of exposure. AB - STUDY OBJECTIVE: We set the hypothesis that follow-up surveys of occupational asthma (OA) could now show better improvement in the asthmatic condition because of a more prolonged interval since removal from exposure than in previously reported studies. PATIENTS/METHODS: Ninety-nine subjects with OA were assessed and were separated into two groups according to the duration of cessation of exposure: (1) group removed for > or = 5 years: 48 subjects studied 8.9+/-2.2 years after cessation of exposure; (2) group removed for <5 years: 51 subjects with OA, comparable in terms of history and functional results at time of diagnosis, with a time lapse from last exposure of 3.1+/-1.2 years. On the follow up visit, questionnaires including information on the current and previous use of inhaled steroids, spirometry, and methacholine tests were administered and results were compared with those obtained at the time of diagnosis. RESULTS: At the follow-up visit, no significant changes in spirometry were observed in the two groups. However, a significant improvement in provocative concentration of methacholine causing a 20% fall in FEV1 (PC20) from a mean value of 1.5 to 3.7 mg/mL was documented (p<0.001). The proportion of subjects having normal PC20 at the follow-up visit was significantly higher in the group removed from exposure for >5 years than in the group removed for < or = 5 years (16/33 vs 8/42; p=0.01). Stepwise logistic regression showed that follow-up PC20 could be predicted from baseline PC20 (p<0.001, odds ratio [OR]=4.1, 95% confidence interval [CI]=1.8 to 9.1), duration of exposure (p=0.04, OR=0.9, 95% CI=0.8 to 1.0), the interval since removal from exposure (p=0.002, OR=1.7, 95% CI=1.2 to 2.5), and the type of agent; subjects with OA due to high-molecular-weight agent showed a less favorable outcome (p=0.04, OR=0.2, 95% CI=0.03 to 1.0). Current and past treatments with inhaled steroids were not significant predictors. CONCLUSION: Results obtained in the group of this study removed for >5 years show better prognostic figures than those reported in most previous studies. Comparison with the group removed for a shorter interval and the stepwise logistic regression analysis suggest that the longer duration of the interval from cessation of exposure appears to be a factor determining this difference. PMID- 9726722 TI - Sensitivity of aerosol bolus behavior to methacholine-induced bronchoconstriction. AB - STUDY OBJECTIVES: Airway narrowing causes alterations in the shape of an exhaled aerosol bolus that can serve as indexes of airway changes during bronchoprovocation. We compared the sensitivities of aerosol bolus behavior and specific airway conductance (SGaw) during bronchoprovocation in normal subjects. DESIGN AND PARTICIPANTS: Fifteen normal, nonsmoking subjects were studied. Doubling methacholine (MCh) concentrations were delivered during tidal breathing. After each dose, SGaw was determined followed by inhalation of narrow pulses of 1 microm particles introduced into 1-L breaths. Inhaled and exhaled particle concentrations were measured with light scattering photometry. Using plots of concentration vs volume, the exhaled bolus was compared with the inhaled bolus for measurements of volumetric change in mode location (modal shift), particle deposition, and dispersion. To determine baseline intrasubject variability, sham studies using buffer solution were performed on five subjects. RESULTS: MCh caused a proximal modal shift, and increased dispersion and deposition of the exhaled bolus. At most doses, a greater percentage of subjects showed significant change (p<0.05) from baseline for modal shift and deposition than for SGaw. Aerosol bolus behavior displayed less intrasubject variability than did SGaw during sham studies. CONCLUSION: Aerosol bolus behavior is at least as sensitive as SGaw in detecting MCh-induced airway constriction in normal subjects and exhibits less intrasubject variability. PMID- 9726723 TI - Cardiac effects of formoterol and salmeterol in patients suffering from COPD with preexisting cardiac arrhythmias and hypoxemia. AB - OBJECTIVE: There are several reports of documented adverse cardiac effects during treatment with beta-agonists. Since one should be aware that this may be a problem in patients with preexisting cardiac disorders, we have conducted a randomized, single-blind, balanced, crossover, placebo-controlled study to assess the cardiac effects of two single doses of formoterol (12 microg and 24 microg) and one single dose of salmeterol (50 microg) in 12 patients suffering from COPD with preexisting cardiac arrhythmias and hypoxemia (PaO2<60 mm Hg). DESIGN: Each patient was evaluated at a screening visit that included spirometry, blood gas analysis, plasma potassium measurement, and 12-lead ECG. In following nonconsecutive days, all patients underwent Holter monitoring 24 h during each of the four treatments. Holter monitoring was started soon before drug administration in the morning. Plasma potassium level was measured before drug inhalation, at 2-h intervals for 6 h, and at 9, 12, and 24 h following administration. None of our patients took rescue medication during the 24-h period. RESULTS: Holter monitoring showed a heart rate higher after formoterol, 24 microg, than after formoterol, 12 microg, and salmeterol, 50 microg, and supraventricular or ventricular premature beats more often after formoterol, 24 microg. Formoterol, 24 microg, significantly reduced plasma potassium level for 9 h when compared with placebo, whereas formoterol, 12 microg, was different after 2 h and salmeterol, 50 microg, from 4 to 6 h. CONCLUSIONS: The results of this study suggest that if a COPD patient is suffering from preexisting cardiac arrhythmias and hypoxemia, long-acting beta-agonists may have adverse effects on the myocardium, although the recommended single dose of salmeterol and formoterol, 12 microg, allows a higher safety margin than formoterol, 24 microg. PMID- 9726724 TI - Identification of smokers susceptible to development of chronic airflow limitation: a 13-year follow-up. AB - BACKGROUND: Cigarette smoking is the cardinal cause of COPD, but only a relatively small percentage of smokers have development of clinically overt disease. OBJECTIVES: To identify high-risk subjects and to assess the prognostic significance of "small airways" tests. SETTING: University teaching hospital. SUBJECTS: Fifty-six smokers and ex-smokers of mean age 62.5 years (SD, 2.7) with a smoking history of 40.6 (18.9) pack-years were studied at the end of a 13-year follow-up period. MEASUREMENTS: Questionnaire and lung function tests, including static and dynamic lung volumes, airway resistance, maximal expiratory flow rates, and small airways tests, such as nitrogen slope of the alveolar plateau (N2 slope) and closing volume. RESULTS: Eighty-two percent of subjects with a normal FEV1/vital capacity (VC) ratio at the start of the study (half of them with abnormal results of small airways tests) still had a normal FEV1/VC ratio 13 years later. In the remainder, all but one had final FEV1/VC values >60%. About 80% of subjects with a decreased FEV1/VC at the start (subjects with airflow obstruction) reached at the end of study lower than predicted FEV1/VC values. Only about 10% of these subjects showed an accelerated loss of FEV1, reaching end FEV1/VC values of <45%. Initial N2 slope predicted about 80% of end FEV1 values. CONCLUSION: Middle-aged smokers are at no evident risk of functional deterioration if their FEV1/VC ratio is normal. This is so even if results of small airways tests are abnormal. A decreased FEV1/VC ratio has no serious implications in itself. Only an associated high N2 slope adds the necessary information to predict a low FEV1. Present data suggest that a subgroup of smokers in their 50s, characterized by a low FEV1/VC ratio and a high N2 slope, are probably the susceptible smokers at high risk for development of COPD. PMID- 9726725 TI - Corticosteroids in life-threatening varicella pneumonia. AB - BACKGROUND: Varicella pneumonia that results in respiratory failure or progresses to the institution of mechanical ventilation carries a significant morbidity and mortality despite intensive respiratory support and antiviral therapy. There has been no reported study of the role of corticosteroids in life-threatening varicella pneumonia. DESIGN AND METHODS: This was an uncontrolled retrospective and prospective study of all adult patients with a diagnosis of varicella pneumonia who were admitted to the ICUs of the Johannesburg group of academic hospitals in South Africa between 1980 and 1996. Patient demographics, clinical and laboratory features, necessity for mechanical ventilation, and complications were reviewed. The outcome and therapy of varicella pneumonia was evaluated with particular reference to the use of corticosteroids. Patients with comorbid disease and those already taking immunosuppressive agents were excluded. Key endpoints included length of ICU and hospital stay and mortality. MEASUREMENTS AND RESULTS: Fifteen adult patients were evaluated, six of whom received corticosteroids in addition to antiviral and supportive therapy. These six patients demonstrated a clinically significant therapeutic response. They had significantly shorter hospital (median difference, 10 days; p<0.006) and ICU (median difference, 8 days; p=0.008) stays and there was no mortality, despite the fact that they were admitted to the ICU with significantly lower median ratios between PaO2 and fraction of inspired oxygen than those patients (n=9) who did not receive corticosteroid therapy (86.5 vs 129.5; p=0.045). CONCLUSION: When used in addition to appropriate supportive care and early institution of antiviral therapy, corticosteroids appear to be of value in the treatment of previously well patients with life-threatening varicella pneumonia. The observations presented in this study are important and should form the basis for a randomized controlled trial, as no other relevant studies or guidelines are available. PMID- 9726726 TI - Sequential evaluation of serum adenosine deaminase in patients treated for tuberculosis. AB - STUDY OBJECTIVE: To delineate the course of serum adenosine deaminase (s-ADA) in patients with tuberculosis who are receiving effective therapy. SETTING: A medical ward and an outpatient clinic in a general hospital. PATIENTS: Twenty five immunocompetent patients with pleural or pulmonary tuberculosis. INTERVENTIONS: All patients received standard chemotherapeutic regimens with isoniazid, rifampin, and pyrazinamide. MEASUREMENTS AND RESULTS: Six measurements of several variables, including s-ADA, were carried out at different periods of time during the 6 months of follow-up. There were no significant differences in s ADA values between sexes and there was no significant correlation with age or with the other variables analyzed. There was a significant decline in the s-ADA values during the first 2 months in the patients as a whole (p=0.04), followed by a stabilization of the s-ADA activity. This decline was due to a marked decrease in the s-ADA in the 13 patients (52%) who had initial high levels of the enzyme (p=0.03), whereas there were no changes in those patients with normal initial levels (p=0.27). Patients with increased s-ADA activity at the time of the first measurement reported symptoms for a longer period than patients with normal s-ADA (median, 15 vs 10 days; p=0.02). CONCLUSIONS: s-ADA levels in patients with tuberculosis decrease during the initial months of effective treatment. Perhaps this decrease might reflect the normalization of the altered lymphocyte turnover induced by tuberculosis. The measurement of s-ADA could be of some help to evaluate the response to therapy, particularly in those patients with increased values of the enzyme. PMID- 9726727 TI - Blastomycosis in northeast Tennessee. AB - STUDY OBJECTIVES: To study the epidemiologic and clinical features of blastomycosis in northeast Tennessee. DESIGN: Retrospective review of blastomycosis cases in the region from 1980 through 1995. SETTING: Hospitals located in the Tri-Cities region of northeast Tennessee. PATIENTS: Seventy-two patients with confirmed blastomycosis infection. INTERVENTIONS: None. RESULTS: During the 1980 to 1995 study period, we documented 72 cases of blastomycosis. The mean age was 52 years (range, 13 to 86 years), most were male (69.4%), and nine were immunocompromised. A possible environmental exposure was noted for 28 patients. Pulmonary involvement represented the most common site of infection (61 cases), but multiorgan involvement was common (17 cases). Most patients with pulmonary blastomycosis (66%) presented with a chronic illness, and radiologic findings usually revealed local consolidation or a mass-like lesion. Nine patients developed ARDS with an associated mortality rate of 89%, compared with a 10% mortality for non-ARDS pulmonary cases. Antifungal treatment regimens varied widely, with amphotericin B often used for sicker patients. An epidemiologic evaluation revealed that the mean yearly incidence rate for blastomycosis quadrupled between 1980 and 1987 (0.31 cases/ 100,000 population) and 1988 to 1995 (1.23 cases/100,000 population) (p=0.00001). Most new blastomycosis cases in the 1988 to 1995 period occurred in three counties in the region where significant new construction projects have been underway. CONCLUSION: Blastomycosis is endemic in northeast Tennessee and the number of cases is increasing, coinciding with major new construction in the region. Clinicians in the area must be alert to this condition. PMID- 9726728 TI - Pulmonary infiltrates in neutropenic patients with acute leukemia during chemotherapy: outcome and prognostic factors. AB - STUDY OBJECTIVE: To determine predictors of mortality from pulmonary infiltrates in neutropenic patients with acute leukemia during chemotherapy, and the significance of those factors related to the underlying malignancy and its therapy as well as of those related to the severity of the illness associated with pulmonary infiltrates. DESIGN: A historical cohort study. SETTING: A university teaching hospital and tertiary referral center. PATIENTS AND METHODS: Overall, 53 patients with neutropenia during chemotherapy and with first episodes of pulmonary infiltrates during a 4-year period were studied. Prognostic analysis included 38 variables. Multivariate analyses were performed by logistic regression. RESULTS: The survival rate from pneumonia was 57% (30/53). The following eight parameters were significantly associated with death in univariate analysis: comorbidity present; development of "late" pulmonary infiltrates (> or = 14 days after hospital admission); heart rate > or = 100 beats/min; a ratio heart rate/systolic blood pressure (HR/SBP) > or = 1.2; urea nitrogen > 7 mmol/L; radiographic score > or = 3; neutropenia < 1.0x10(9)/L at the treatment end point; and failed complete remission. In a multivariate model including only parameters available at diagnosis of pulmonary infiltrates, the presence of a ratio HR/SBP > or = 1.2 and of a radiographic score > or = 3 remained independently associated with death. In a second model also including the evolutionary parameter neutropenia < or = 1.0x10(9)/L at the treatment end point, both parameters remained significant together with neutropenia <1.0x 10(9)/L at the treatment end point. The presence of a ratio HR/SBP > or = 1.2 was a strong marker of early death. CONCLUSION: Both therapy- and malignancy-associated neutropenia as well as the severity of illness associated with pulmonary infiltrates are independent prognostic factors. Patients with a ratio HR/SBP > or = 1.2 at diagnosis of pulmonary infiltrates suffer from potentially reversible acute illness, are at risk for early death and, therefore, may be appropriate candidates for treatment in an ICU. PMID- 9726729 TI - Respiratory nitric oxide levels in experimental human influenza. AB - BACKGROUND: Exhaled oral nitric oxide (NO), a reported marker of inflammation in the respiratory tract, can be elevated by "upper respiratory tract infections." However, the responsible viruses and the time course of this rise in NO are not clear. OBJECTIVE: To determine the expired nasal and oral NO levels during experimentally induced influenza A infection in 14 healthy volunteers without a history of asthma, rhinitis, or sinusitis. METHODS: After being housed in individual rooms, susceptible volunteers were inoculated with 10(6) 50% tissue culture infective dose of influenza A/Texas/36/91/(H1N1) virus on a single occasion by intranasal drops. Volunteers remained in the isolation unit for 8 days and returned for follow-up 21 days after inoculation. Symptom scores and nasal washing for viral culture were obtained daily. NO samples from the mouth and nose were obtained on days 0 through 4, 8, and 21 by having the patient perform a slow vital capacity maneuver through a plastic tube into a Mylar balloon. RESULTS: All patients had influenza virus cultured from nasal washings (12 of 14 on day 1, 14 of 14 by day 5). Patient symptom scores peaked on day 3 (mean+/-SE; 15.4+/-3.2) and returned to baseline by day 8. Preinfection exhaled nasal NO (right, 28.4+/-3.7 parts per billion [ppb]; left, 27.7+/-4.6 ppb) was significantly higher than oral NO (5.8 ppb; p<0.001). Exhaled oral NO was significantly elevated on day 8 postinoculation (12.9+/-0.8 ppb; p<0.01 Bonferroni) and returned to baseline at follow-up. Nasal NO levels showed a slight decrease on days 2 to 4 but returned to baseline by day 8. CONCLUSION: Experimental influenza virus infection can increase oral but not nasal exhaled NO levels. The timing of exhaled NO changes suggests that NO does not contribute to illness manifestations directly. PMID- 9726730 TI - Maximum cardiac output during incremental exercise by first-pass radionuclide ventriculography. AB - STUDY OBJECTIVE: To validate a noninvasive first-pass radionuclide ventriculographic (FPRV) measurement of maximum cardiac output (Qv) during exercise. DESIGN: Comparison of Qv to that measured by the Fick principle (Qf) at peak exercise. SETTING: Academic cardiopulmonary exercise laboratory. PATIENTS: Seventy-eight consecutive patients without a history of septal defect undergoing clinically indicated maximum incremental cardiopulmonary exercise testing with pulmonary arterial catheterization and FPRV. MEASUREMENTS AND RESULTS: Ventilation and gas exchange were measured breath-by-breath or by a mixing chamber/mass spectrometer system. Arterial and mixed venous O2 content were measured each minute during exercise. When patients without left-to-right ventricular stroke count ratio evidence for left-sided regurgitation were isolated, peak Qv was linearly related to Qf (r=0.75, p=0.0001). To account for a small systematic overestimation (bias) of Qf by Qv, the linear equation for the Qv/Qf relation was derived for patients studied between 1990 and 1993 and applied to those studied subsequently. The resulting corrected peak Qv was tightly related to peak Qf (r=0.90, p<0.001) with confidence intervals for slope and intercept overlapping identity. CONCLUSION: FPRV can reasonably estimate maximum cardiac output during incremental exercise in patients for whom the technique has ruled out left-sided cardiac regurgitant lesions. PMID- 9726731 TI - The new onset of atrial arrhythmias following major noncardiothoracic surgery is associated with increased mortality. AB - STUDY OBJECTIVES: To examine the incidence and consequences of atrial arrhythmias in surgical ICU patients following major noncardiac, nonthoracic surgery. DESIGN: Prospective observational study. SETTING: University hospital surgical ICU. PATIENTS: Four hundred sixty-two consecutive patients after noncardiothoracic surgery. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: Patients were assigned to one of three groups: group 1-new-onset atrial arrhythmias (n=47); group 2-history of atrial arrhythmias (n=58); and group 3-no atrial arrhythmias (n=357). New arrhythmias occurred in 10.2% of patients. Most began within the first 2 postoperative days. These patients had a higher mortality rate (23.4%), longer ICU stay (8.5+/-17.4 [SD] days), and extended hospital stay (23.3+/-23.6 days) than patients without atrial arrhythmias (mortality, 4.3%; ICU stay, 2.0+/-4.5 days; hospital stay; 13.3+/-17.7 days; p<0.02). Thirteen percent of patients had a history of atrial arrhythmias. They had a higher mortality rate (8.6%) and longer ICU stays (2.9+/-4.9 days; p<0.02) than patients without arrhythmias. Most deaths in the two arrhythmia groups were not due to cardiac problems, but to sepsis or cancer. CONCLUSIONS: Patients admitted to a surgical ICU after noncardiothoracic surgery with a history of or who developed new atrial arrhythmias had greater mortality and longer ICU stays than patients without arrhythmias. The incidence of new-onset arrhythmias was lower than reported after cardiac and thoracic surgery, but higher than in the general population. Atrial arrhythmias were not the cause of death and appear to be markers of increased mortality and morbidity. PMID- 9726732 TI - Transient bradycardia induced by carotid sinus pressure increases outflow obstruction in hypertrophic obstructive cardiomyopathy but not in valvular aortic stenosis. AB - BACKGROUND: The murmur of hypertrophic obstructive cardiomyopathy (HOCM) increases in intensity in about 80% of those patients in whom carotid sinus pressure (CSP) slows the heart rate. This does not occur in valvular aortic stenosis (AS). STUDY OBJECTIVES, DESIGN, AND PATIENTS: It was hypothesized that left ventricular (LV) obstruction increases with CSP in HOCM and not in AS. Furthermore, it was not clear whether it was the sudden bradycardia or CSP itself that was responsible for the effect noted. Therefore, studies were performed using two different interventions: (1) Doppler echocardiography was performed before and during CSP in 36 HOCM patients and 21 AS patients; (2) two patients with DDD pacemakers and HOCM were examined before and after pacemaker rate slowing. Finally, atrial pacing was performed in three HOCM patients at catheterization, and atrial pacing was either slowed or stopped (without CSP). RESULTS: LV outflow velocity and pressure gradient increased in 28 of 30 HOCM patients (92%) in whom heart rate decreased with CSP. The peak instantaneous pressure gradient increased from 45+/-37 to 77+/-53 mm Hg (p<0.005), and the velocity contour became more typical of HOCM. The pressure gradient increased from 30 mm Hg to 64 and 81 mm Hg, respectively, in the two patients with DDD pacemakers after pacemaker rate slowing. Similar results were seen with slowing or cessation of atrial pacing at catheterization. In contrast, the pressure gradient increased in only three of 21 AS patients (14%), to 44+/-28 from 41+/-25 mm Hg, and remained unchanged in the other 18. CONCLUSION: This study shows that LV outflow velocity and pressure gradient increase markedly in most HOCM patients (92%) if CSP succeeds in slowing the heart rate, but not in patients with valvular AS. A similar effect is obtained by simply decreasing the atrial rate in patients with DDD or atrial pacemakers. This increase in outflow tract obstruction is sufficient to account for the increase in murmur intensity. Decreased afterload (secondary to greater aortic decompression with the longer diastole), increased intrinsic force of contraction with the bradycardia (the Woodworth effect), and Starling's law may play independent roles in the dynamic increase in obstruction observed during CSP in patients with HOCM. Worsening of mitral regurgitation was not clearly shown to contribute to the increase in murmur, but it cannot readily be ruled out. PMID- 9726733 TI - Assessment of left ventricular diastolic function after single lung transplantation in patients with severe pulmonary hypertension. AB - OBJECTIVES: This study was designed to observe left ventricular filling by Doppler echocardiography before and after single lung transplantation in patients with severe pulmonary hypertension. BACKGROUND: Right ventricular pressure overload causes the deformation of the left ventricle by septal flattening toward its cavity, which may result in impaired left ventricular early filling. Recent studies have demonstrated the ability of single lung transplantation to restore right ventricular function in patients with severe pulmonary hypertension. However, changes in left ventricular filling after single lung transplantation have not been well studied. METHODS: We performed Doppler echocardiography in nine patients with severe pulmonary hypertension before, early (<3 months), and late (>1 year) after single lung transplantation. The study group consisted of eight female patients and one male patient with mean age of 32 years (range, 15 to 48 years). Six patients were diagnosed as having primary pulmonary hypertension and three as having secondary pulmonary hypertension. Nine age matched normal subjects served as a control group. Doppler measurements included the following: transmitral flow early (E) and atrial (A) velocities, integrals (Ei and Ai), and left ventricular isovolumic relaxation time. The ratio of E/A and atrial filling fraction (Ai/Ei+Ai, AFF) were also determined. Left ventricular geometry was assessed from mid-short axis view with a circular shape factor (CSF). RESULTS: Early after lung transplantation, the left ventricular geometry became more circular with CSF (mean+/-SD) increasing from 0.63+/-0.09 to 0.88+/-0.05 (p<0.05). However, impaired early filling persisted in the patient group (E/A 0.7+/-0.1 vs preoperative 0.6+/-0.1, AFF 0.61+/-0.1 vs 0.64+/-0.1; both p=not significant). One year later, the left ventricular filling had returned to normal range with E/A 1.4+/-0.6 and AFF 0.35+/-0.1. CONCLUSIONS: This study observed that the impaired left ventricular early filling persisted shortly after single lung transplantation in patients with severe pulmonary hypertension, despite findings that left ventricular geometry was restored earlier after reversal of pulmonary hypertension. The abnormal filling pattern appeared to be resolved 1 year later. The findings suggest the impaired early filling may be caused by intrinsic left ventricular abnormalities other than ventricular interaction in these patients. PMID- 9726734 TI - Troponin I, troponin T, or creatine kinase-MB to detect perioperative myocardial damage after coronary artery bypass surgery. AB - STUDY OBJECTIVES: To compare cardiac troponin I (cTnI), cardiac troponin T (cTnT), and creatine kinase MB (CKMB mass) in patients with and without new Q wave on the ECG following coronary artery bypass graft (CABG) surgery. PATIENTS: After ethic committee's approval and informed consent, 82 patients, mean age 63+/ 10 years, scheduled for CABG were included. INTERVENTIONS: Arterial blood samples were drawn during cardiopulmonary bypass, before, and 6, 12, 24, and 48 h after aortic cross-clamp release. cTnI, cTnT, and CKMB mass were measured. The appearance of new Q wave on the ECG performed preoperatively and 24 h postoperatively was used to assess myocardial lesion independently of biological markers. RESULTS: There were 69 patients without new Q wave on the ECG (group 1) and 13 with (group 2). In group 1, cTnI reached a peak of 2.1 microg/L (median, interquartile range [IQ]=2.4) at 12 h, cTnT increased progressively with a peak of 0.22 microg/L (IQ=0.2) at 48 h, and CKMB presented an earlier peak of 10 microg/L (IQ=6.2) at 6 h. Starting with the same median value, group 2 patients presented significantly higher peaks: cTnI: 17 microg/L (IQ=16) at 12 h; cTnT: 1.4 microg/L (IQ=2.3) at 12 h; and CKMB mass: 74 microg/L (IQ=61) at 6 h. Receiver operating characteristic (ROC) curves were constructed. The area under the curve was 0.90 for cTnI, 0.84 for CKMB, and 0.81 for cTnT (not significant). The best cutoff values to discriminate between group 1 and group 2 patients were determined with the ROC curves: cTnI=5 microg/L; CKMB mass=20 microg/L; cTnT=0.3 microg/L. Sensitivity, specificity, and positive and negative values for cTnI (5 microg/L) were 91%, 82%, 53%, and 98%, respectively. CONCLUSIONS: There was little differences among cTnI, cTnT, and CKMB after CABG to diagnose myocardial damage as assessed by new Q wave on the ECG. There was a trend of cTnI to be a better discriminator than cTnT, but it did not reach statistical significance. PMID- 9726735 TI - The effects of preoperative therapy with angiotensin-converting enzyme inhibitors on clinical outcome after cardiovascular surgery. AB - OBJECTIVE: To determine the effect of preoperative therapy with angiotensin converting enzyme (ACE) inhibitors on clinical outcome after cardiovascular surgery. STUDY: Inception cohort. SETTING: A tertiary care 54-bed cardiothoracic ICU. PATIENTS: All admissions to an ICU over a 42-month period after cardiovascular surgery. INTERVENTION: Extraction of preoperative, operative, and ICU data from a database. OUTCOME MEASURES: Incidence of acute organ dysfunction, length of mechanical ventilation, ICU stay, and death after cardiovascular surgery. RESULTS: The study cohort consisted of four groups: normal or moderately impaired left ventricular function control (group A, n=6,400); normal or moderately impaired left ventricular function treated with ACE inhibitors (group B, n=1,375); severe left ventricular dysfunction control (group C, n=1,905); and severe left ventricular dysfunction treated with ACE inhibitors (group D, n=1,650). The incidence of three or more organ dysfunction was similar on comparison of group A vs group B (5% vs 6%) or group C vs group D (15% vs 13%). There were no differences in the total duration of mechanical ventilation or length of stay in the ICU in group A vs group B or group C vs group D. Death occurred in 2% of groups A and B, and at 6% in groups C and D. Preoperative severe left ventricular dysfunction in both groups C and D was associated with an increased incidence of three or more organ dysfunction, duration of mechanical ventilation, length of stay in ICU, and death after surgery. Multivariate analysis indicated that therapy with ACE inhibitors did not affect the clinical outcome after cardiovascular surgery. CONCLUSION: Preoperative therapy with ACE inhibitors did not influence the clinical outcome after cardiac surgery. It is unlikely that therapy with ACE inhibitors can alter the clinical sequelae of cardiopulmonary bypass and cardiac surgical procedures performed in high-risk patients because of underlying severe left ventricular dysfunction. PMID- 9726736 TI - Bronchial responsiveness and angiotensin-converting enzyme gene polymorphism in sarcoidosis patients. AB - BACKGROUND: Angiotensin-converting enzyme (ACE) inactivates bradykinin and tachykinins, which are potent bronchoconstrictors and mediators of inflammatory reactions. It has recently been shown that an insertion (I)/deletion (D) polymorphism in the ACE gene accounts for variation in serum ACE level. We investigated bronchial responsiveness in patients with sarcoidosis to determine whether it might be associated with ACE gene polymorphism. SUBJECTS: Bronchial responsiveness was assessed in 21 patients with sarcoidosis, 21 patients with asthma, and 18 healthy control subjects. ACE polymorphism was also examined in the 21 patients with sarcoidosis. METHODS: Bronchial responsiveness was measured by recording respiratory resistance with continuous inhalation of methacholine from 49 to 25,000 microg/mL in concentration. The ACE genotype was determined using the polymerase chain reaction. RESULTS: We found a significant increase in bronchial responsiveness in sarcoidosis patients as compared with healthy control subjects (p<0.01). In the sarcoidosis group, patients with the II genotype demonstrated significantly more coughing (p<0.05) and a greater bronchial responsiveness (p<0.05) than did those with DI or DD genotypes. CONCLUSION: Patients with sarcoidosis have increased bronchial responsiveness to some extent, the degree apparently depending on the ACE genotype. PMID- 9726737 TI - Pulmonary Wegener's granulomatosis: correlation between high-resolution CT findings and clinical scoring of disease activity. AB - STUDY OBJECTIVE: To evaluate the usefulness of high-resolution CT (HRCT) for monitoring pulmonary disease activity in Wegener's granulomatosis (WG). DESIGN: Prospective study of CT and clinical data. SETTING: Main referral hospital for rheumatic diseases and department of diagnostic radiology of collaborating university hospital. PATIENTS: Seventy-three patients with WG underwent 98 staging examinations using HRCT. The status of pulmonary disease activity at the time of examination was scored according to clinical, bronchoscopic, BAL, and radiographic findings as follows: activity (n=25, group 1), past activity (n=45, group 2) and lack of any pulmonary disease (n=28, group 3). HRCT findings were correlated with the clinical scoring of pulmonary disease activity. RESULTS: Of 98 staging examinations 78 (79.6%) revealed abnormal CT scans showing the following main abnormalities: (a) nodules or masses (group 1: 16 [60.4%], group 2: 9 [20%]); (b) parenchymal bands (group 1: 12 [48%], group 2: 27 [60%], group 3: 6 [21.5%]); (c) septal thickening (group 1: 8 [32%], group 2: 6 [13.3%]); (d) parenchymal opacification (group 1: 7 [28%], group 2: 4 [8.9%]); and (e) pleural irregularity (group 1: 14 [56%], group 2: 22 [49%], group 3: 9 [32%]). Nodules/masses and areas of parenchymal opacification were significantly associated with florid disease activity of the lungs. Parenchymal bands and septal thickening were observed in both groups with pulmonary involvement, but statistical analysis revealed no significant difference. Pleural irregularities were nonspecific. CONCLUSION: HRCT may be a useful adjunct to clinical scoring of pulmonary disease activity in patients with WG and suspected lung involvement. PMID- 9726738 TI - Colchicine, D-penicillamine, and prednisone in the treatment of idiopathic pulmonary fibrosis: a controlled clinical trial. AB - STUDY OBJECTIVE: We compared the long-term efficacy of the combination of colchicine and/or D-penicillamine with prednisone, in comparison to prednisone alone in patients with idiopathic pulmonary fibrosis (IPF). DESIGN: Nonrandomized prospective study in patients with IPF confirmed by biopsy specimen. SETTING: National Institute of Respiratory Diseases, Mexico. PATIENTS: Fifty-six IPF patients were included in this study. Patients received either colchicine/ prednisone (n=19), D-penicillamine/prednisone (n=11), D penicillamine/colchicine/prednisone (n=11), or prednisone alone (n=15). Prednisone therapy was started at 1.0 mg/kg/d for 1 month followed by a biweekly taper to a maintenance dose of 15 mg/d. Colchicine was administered at a daily dose of 1.0 mg, and D-penicillamine was given at a daily dose of 600 mg. MEASUREMENTS AND RESULTS: Response to therapy was assessed by changes in lung function test results as measured by total and vital lung capacities, arterial blood gas analysis at rest breathing room air, and survival. No significant differences either in lung mechanics or in arterial gases were found in any group relative to the baseline measurement. Thirteen of the 56 patients died during the first 2 years, and 29 were dead at 5 years follow-up. Comparison of survival curves by Cox regression model showed no statistically significant difference among the four groups. Known side effects attributable to prednisone were more common and severe than those attributable to the other drugs. CONCLUSIONS: Our results suggest that neither colchicine nor D-penicillamine modified the progressive course of prednisone-treated IPF, and that the search for new drugs is imperative. PMID- 9726739 TI - The validity of radiographic estimation of total lung capacity in patients with respiratory disease. AB - STUDY OBJECTIVE: To evaluate the validity of a state-of-the-art computerized planimetry technique for estimation of total lung capacity (TLC) from chest radiographs, when applied to patients with clinical lung disease receiving routine chest radiographs. DESIGN: Retrospective clinical survey. SETTING: An occupational medicine diagnostic clinic. PATIENTS: A convenience sample of 40 subjects with asbestos-related lung disease, 5 patients with nonasbestos-related restrictive defects, 15 subjects with occupational asthma, and 10 subjects with irritant tracheobronchitis. RESULTS: Estimation of TLC using state-of-the-art computerized algorithms demonstrated limited agreement with conventional measures of TLC when applied to patients with occupational lung disease receiving routine chest radiographs. The most pronounced differences occurred in patients with asbestos-related lung disease and restrictive defects, where the radiographic method of measurement significantly overestimated helium dilution TLC by 986 mL (r=0.73, p<0.001) and 1,135 mL (r=0.82, p<0.05), respectively. Good inspiratory effort was associated with significantly increased radiographic TLC relative to helium dilution TLC; however, radiographic features did not fully account for the observed differences between radiographic and helium dilution techniques. CONCLUSIONS: Our findings suggest that this planimetric technique should not be used as a substitute for conventional measures of TLC in clinic populations receiving routine radiographs. The large diagnostic group specific mean differences observed between radiographic and conventional measures of TLC also suggest that this method is of limited utility in clinical evaluation of occupational lung disease. PMID- 9726740 TI - Long-term within-subject variability of inspiratory neural drive response to hypoxia. AB - STUDY OBJECTIVE: We analyze the within-subject variation of mouth occlusion pressure (P0.1) response to progressive isocapnic hypoxic stimulation over long time periods in normal subjects. PATIENTS AND INTERVENTIONS: We studied 21 healthy subjects (14 male and 7 female; aged 40+/-12 yrs) (mean+/-SD). Lung volumes, basal P0.1, and P0.1 response to hypoxia were measured on two separate occasions 2 months apart, under similar ambient and clinical conditions. RESULTS: There was no significant change in clinical condition, FVC, FEV1, arterial oxygenation saturation, end-tidal and mixed venous PCO2 levels, or P0.1 between the two visits. The mean P0.1 responses to hypoxia in the two explorations were 0.032+/-0.022 and 0.034+/-0.022 kPa/%, respectively. There was a moderate intrasubject variability of P0.1 response to hypoxia, with a coefficient of reproducibility of 0.01 kPa/%. Power calculations to establish the optimal sample size required for hypoxic stimulation are presented. CONCLUSION: Long term within subject variability of P0.1 response to hypoxia is moderate. This intrinsic variability needs to be emphasized when interpreting the effects of experimental interventions on hypoxic sensitivity. PMID- 9726741 TI - Pulmonary metastases of endocrine origin: the role of surgery. AB - PURPOSE: To determine the clinical course and outcome of patients undergoing pulmonary resection for metastatic endocrine tumors. METHODS: Retrospective review of 47 patients with known endocrine tumors and pulmonary metastases who were evaluated for surgical resection between 1975 and 1996. RESULTS: Tumors evaluated included the following: carcinoid (16), thyroid (12), pancreatic adenocarcinoma (10), adrenocortical carcinoma (6), pheochromocytoma (2), and parathyroid (1). Thirty-three patients were asymptomatic. Hormone secretion was noted in five patients. Twenty-five patients, who had isolated lung metastases, good control of the primary tumor, and no medical contraindication had surgical resection. The number of pulmonary nodules was not a limiting factor as long as all disease could be resected with adequate residual pulmonary function. CT was successful in directing resection in all patients. Twenty-six operations were performed in 25 patients and 22 patients were treated medically. Wedge resection was performed for lesions <2 cm (15), and lobectomy for larger or multiple nodules (10). Four patients had bilateral nodules resected. There was no operative mortality and no major complications. Actuarial 5-year survival was 61% for surgically treated patients. Independent predictors of poor survival included positive mediastinal lymph nodes at time of surgery (p=0.004) and shorter disease free interval (p=0.01). At a median of 6.7+/-1.2 years, six patients have developed radiographic appearance of a recurrence. A single patient with recurrent Hurthle cell cancer has had a successful reresection. The remaining patients have received chemotherapy. No patient with pancreatic carcinoma or adrenocortical carcinoma was a candidate for resection. All medically treated patients died within 6 months. CONCLUSION: Patients with endocrine tumors and pulmonary metastases are usually asymptomatic, their conditions are diagnosed accurately with CT, and they can achieve long-term survival comparable to other tumors (sarcoma) after pulmonary metastasectomy. CLINICAL IMPLICATIONS: Patients with carcinoid, thyroid, pheochromocytoma, and parathyroid tumors with pulmonary metastases should undergo surgical resection if there is the following: (1) no evidence of extrathoracic disease; (2) good control of the primary tumor; (3) no medical contraindications for surgery; and (4) pulmonary function that can tolerate resection of all documented disease. The role of adjuvant chemotherapy in patients with positive lymph nodes needs further study. PMID- 9726742 TI - Identification and modification of environmental noise in an ICU setting. AB - STUDY OBJECTIVES: Noise levels in the hospital setting are exceedingly high, especially in the ICU environment. We set out to determine what caused the noises producing sound peaks > or = 80 A-weighted decibels (dBA) in our ICU settings, and attempted to reduce the number of sound peaks > or = 80 dBA through a behavior modification program. DESIGN: The study was divided into two separate phases: noise identification and a trial of behavior modification. During the noise identification phase we simultaneously recorded sound peaks and the loudest noise heard subjectively by one observer in the medical ICU (MICU) and the respiratory ICU (RICU). During the behavior modification phase of the study we implemented a behavior modification program, geared toward noise reduction, in all of the MICU staff. Sound levels were monitored before and at the end of the behavior modification trial. SETTING: The MICU and RICU of a 720-bed teaching hospital in Providence, RI. PARTICIPANTS: All ICU staff during the study period. INTERVENTIONS: Once the noises that were determined to be amenable to behavior modification were identified, a behavior modification program was conducted during a 3-week period in our MICU. Baseline and post-behavior modification noise recordings were compared in 6-h intervals after sites were matched by number of patients in a room and Acute Physiology and Chronic Health Evaluation II (APACHE II) scores. MEASUREMENTS AND RESULTS: We identified several causes of sound peaks > or = 80 dBA amenable to behavior modification; television and talking accounted for 49%. We also significantly reduced the 24-h mean peak noise level (p=0.0001), as well as the mean peak noise level (p=0.0001) and the number of sound peaks > or = 80 dBA (p=0.0001) in all 6-h blocks except for the 12 AM to 6 AM period. CONCLUSIONS: We conclude that many of the noises causing sound peaks > or =80 dBA are amenable to behavior modification and that it is possible to reduce the noise levels in an ICU setting significantly through a program of behavior modification. PMID- 9726743 TI - The use of continuous i.v. sedation is associated with prolongation of mechanical ventilation. AB - STUDY OBJECTIVE: To determine whether the use of continuous i.v. sedation is associated with prolongation of the duration of mechanical ventilation. DESIGN: Prospective observational cohort study. SETTING: The medical ICU of Barnes-Jewish Hospital, a university-affiliated urban teaching hospital. PATIENTS: Two hundred forty-two consecutive ICU patients requiring mechanical ventilation. INTERVENTIONS: Patient surveillance and data collection. MEASUREMENTS AND RESULTS: The primary outcome measure was the duration of mechanical ventilation. Secondary outcome measures included ICU and hospital lengths of stay, hospital mortality, and acquired organ system derangements. A total of 93 (38.4%) mechanically ventilated patients received continuous i.v. sedation while 149 (61.6%) patients received either bolus administration of i.v. sedation (n=64) or no i.v. sedation (n=85) following intubation. The duration of mechanical ventilation was significantly longer for patients receiving continuous i.v. sedation compared with patients not receiving continuous i.v. sedation (185+/-190 h vs 55.6+/-75.6 h; p<0.001). Similarly, the lengths of intensive care (13.5+/ 33.7 days vs 4.8+/-4.1 days; p<0.001) and hospitalization (21.0+/-25.1 days vs 12.8+/-14.1 days; p<0.001) were statistically longer among patients receiving continuous i.v. sedation. Multiple linear regression analysis, adjusting for age, gender, severity of illness, mortality, indication for mechanical ventilation, use of chemical paralysis, presence of a tracheostomy, and the number of acquired organ system derangements, found the adjusted duration of mechanical ventilation to be significantly longer for patients receiving continuous i.v. sedation compared with patients who did not receive continuous i.v. sedation (148 h [95% confidence interval: 121, 175 h] vs 78.7 h [95% confidence interval: 68.9, 88.6 h]; p<0.001). CONCLUSION: We conclude from these preliminary observational data that the use of continuous i.v. sedation may be associated with the prolongation of mechanical ventilation. This study suggests that strategies targeted at reducing the use of continuous i.v. sedation could shorten the duration of mechanical ventilation for some patients. Prospective randomized clinical trials, using well-designed sedation guidelines and protocols, are required to determine whether patient-specific outcomes (eg, duration of mechanical ventilation, patient comfort) can be improved compared with conventional sedation practices. PMID- 9726744 TI - Utility of transbronchial biopsy in patients with acute respiratory failure: a postmortem study. AB - STUDY OBJECTIVE: To determine the diagnostic yield of histologic specimens obtained by postmortem transbronchial biopsy (TBB) in patients with acute respiratory failure requiring mechanical ventilation. DESIGN: Standard postmortem histologic examination of lung tissue specimens. SETTING: An urban university affiliated hospital. PATIENTS OR PARTICIPANTS: Thirty patients with diffuse pulmonary infiltrates and acute respiratory failure, who underwent postmortem examination. INTERVENTIONS: Following removal of the lungs from the thorax. TBBs were obtained from the lower lobe of each deflated lung and comparison was made to a 1-cm3 tissue block obtained from the ipsilateral lower lobe. MEASUREMENTS AND RESULTS: Standard postmortem histologic examination provided a specific diagnosis in 85% of the 60 lungs examined, and histologic evidence of acute pneumonia was present in 30% of the lungs. The overall yield of TBB was 48% for establishing a specific histologic diagnosis and 15% for the diagnosis of acute pneumonia. Using standard postmortem histologic examination as the gold standard, the sensitivity and specificity of TBB for making a specific diagnosis were 57% and 100% respectively, with corresponding positive and negative predictive values of 100% and 29%. For the histologic diagnosis of acute pneumonia, the sensitivity of TBB was 50%, the specificity was 100%, and the positive and negative predictive values were 100% and 82%, respectively. The kappa statistic for the agreement between the two diagnostic methods was 0.28 for establishing a specific diagnosis and 0.58 for the diagnosis of acute pneumonia. Obtaining 12 TBBs rather than six TBBs did not increase the diagnostic yield for TBB. CONCLUSIONS: These findings suggest poor overall agreement between standard postmortem histologic examination and TBB specimens. Although not performed in a clinical setting, this postmortem investigation suggests that TBB may be of limited value in mechanically ventilated patients with acute respiratory failure because of its low sensitivity. PMID- 9726745 TI - Does positive end-expiratory pressure ventilation improve left ventricular function? A comparative study by transesophageal echocardiography in cardiac and noncardiac patients. AB - STUDY OBJECTIVES: Positive end-expiratory pressure (PEEP) has been proposed to improve cardiac output in patients with left ventricular (LV) dysfunction. This study was designed to compare quantitative global and regional LV performance in response to PEEP in patients with normal and poor LV function. DESIGN: A prospective clinical trial. SETTING: Adult medical ICU in a university hospital. PATIENTS: Twelve critically ill patients requiring respiratory support and divided into two groups according to baseline transesophageal echocardiographic (TEE) measurements: normal LV dimensions and fractional area of contraction (FAC=61+/-5%) (n=7) and dilated cardiomyopathy with reduced FAC (21+/-1%) (n=5). MEASUREMENTS AND RESULTS: All patients were studied when two successive levels of PEEP (best PEEP as the highest value of respiratory compliance and high PEEP as best PEEP+10 cm H2O) were applied. Global systolic LV performance and quantitative regional wall motion analysis performed by the centerline method were assessed on the TEE transgastric short-axis view. End-systolic wall stress (ESWS) was used as a reliable indication of LV afterload. PEEP reduced LV dimensions asymmetrically in both groups of patients and septolateral diameter significantly decreased without affecting global LV systolic performance. Additionally, high PEEP produced a significant impairment in septal kinetics as evidenced by the centerline method. High PEEP also decreased ESWS for all patients (-27% in normal group and -23% in cardiac group, p<0.05) without significant improvement in global systolic LV performance (FAC: +2% in normal group and +0% in cardiac group; not significant). CONCLUSIONS: PEEP cannot be recommended routinely to improve LV performance in patients with severe dilated cardiomyopathy. PMID- 9726746 TI - Doxycycline pleurodesis in rabbits: comparison of results with and without chest tube. AB - We have reported previously that there is a high incidence of hemothorax and substantial mortality in rabbits that are given tetracycline derivatives intrapleurally. However, such complications have not been reported in humans when pleurodesis is attempted with tetracycline derivatives. One primary difference in the two situations is that a chest tube is placed only in humans. The objective of this study was to evaluate the hypothesis that chest tube placement would prevent the development of hemothoraces and lead to better pleurodesis in rabbits given doxycycline intrapleurally. Eighty New Zealand White male rabbits received doxycycline, 20 mg/kg, in a total volume of 2 mL. One half of the rabbits were randomized to receive a chest tube at the time of the injection and were subjected to pleural fluid aspiration twice daily. The remaining rabbits (control group) received no chest tube and no aspiration. Ten rabbits from each group were killed on days 4, 7, 14, and 28. The intrapleural injection of doxycycline induced the production of large exudative effusions. The insertion of chest tubes prevented the development of hemothorax (0/20 in chest tube group, 15/20 in control group, p<0.001). The insertion of chest tubes was also associated with a significant reduction in mortality and a significant improvement in pleurodesis. When pleurodesis is attempted in rabbits with intrapleural doxycycline, the insertion of a chest tube will prevent hemothorax and lead to a better pleurodesis. PMID- 9726747 TI - Intrathoracic and extrathoracic sources of exhaled nitric oxide in porcine endotoxemic shock. AB - OBJECTIVES: Nitric oxide (NO), a highly reactive species produced by the activity of NO synthases (NOS), is normally present in the exhaled air of humans and animals. Exhaled NO concentration increases significantly in humans with sepsis and animals, but neither the source nor NOS isoforms responsible for this rise in pulmonary NO production are known. The main objective of this study is to determine the sites and the mechanisms of enhanced NO production in the exhaled air of endotoxemic pigs. DESIGN: Randomized, controlled, animal study. SETTING: University-based animal research facility. SUBJECTS: Thirteen pathogen-free adult female pigs (22 to 27 kg). INTERVENTIONS: Anesthetized pigs were divided into two groups: control and lipopolysaccharides (LPS) (septic) groups. In both groups, extrathoracic (upper airways, nasal, and paranasal) and intrathoracic (bronchi, bronchioles, and alveoli) compartments were ventilated equally with two separate ventilators connected to two tracheal tubes. The LPS group received slow infusion (over 2 h) of Escherichia coli endotoxin (10 microg/kg/h), whereas saline solution was infused into the control group. Expired air of the two compartments was collected throughout the 2-h observation period. The animals were then killed and the lungs were quickly excised and frozen. MEASUREMENTS: Hemodynamic variables were measured in both groups. NO concentration in the exhaled air of both compartments was measured with a chemiluminescence analyser. Pulmonary NOS activity was evaluated by measuring the conversion of L-[2,3H]-arginine to L [2,3H]-citrulline, and pulmonary expression of NOS was evaluated by immunoblotting. RESULTS: Baseline NO concentration in both groups was significantly higher in the extrathoracic vs intrathoracic compartment (average of 5.2 vs 3.4 parts per billion). Endotoxin infusion elicited a significant and early (after 45 min) rise in exhaled NO concentration in the extrathoracic compartment. Exhaled NO in the intrathoracic compartment also rose significantly but after 90 min of endotoxin infusion. Measurement of lung NOS activity showed a substantial rise in Ca++/calmodulin-dependent activity in the LPS group with no rise in Ca++/calmodulin-independent activity. Immunoblotting of lung tissue samples indicated the absence of the inducible isoform in both groups of animals. Moreover, LPS injection elicited no significant alterations in the pulmonary expression of the endothelial and the neuronal isoforms. CONCLUSIONS: Both extrathoracic and intrathoracic compartments contribute to the rise in exhaled NO production in experimental septic shock. The rise in exhaled NO production is due to increased activity of constitutive NOS isoforms as a result of increased cofactor availability and/or downregulation of the endogenous inhibitors of NOS. PMID- 9726748 TI - Effects of repetitive use and cleaning techniques of disposable jet nebulizers on aerosol generation. AB - STUDY OBJECTIVE: Patients with cystic fibrosis use disposable jet nebulizers for the self-administration of antibiotics, DNase, and bronchodilators several times per day. Most patients elect to reuse their disposable nebulizers. The purpose of this study was to determine if significant changes in particle size distribution or output (mL/min) occurred with reuse. DESIGN: In vitro studies were performed using four disposable models and one durable jet nebulizer for up to 100 runs; measurements of particle size and output were obtained at 10 run intervals, using saline solution alone, tobramycin, gentamicin, or a mixture of albuterol and cromolyn. Particle size determinations were made with a laser diffraction analyzer. RESULTS: There was no significant difference between the baseline performance of the four disposable models and the durable Pari LC, when measuring particle size distribution of the aerosol; the Pari LC had an output rate two to three times higher than the four disposable models. For each of the four solutes tested, there was no clinically significant change in performance for up to 100 cycles, when the nebulizers were properly cleaned between uses. Unwashed units containing tobramycin started to fail by 40 runs. CONCLUSIONS: When properly maintained, there was no trend of deterioration of performance with repeated use of disposable nebulizers. Microbial contamination was not addressed in this study and must be considered prior to recommendations for the reuse of disposable nebulizers. PMID- 9726749 TI - Selection of peak flowmeters in ambulatory asthma patients: a review of the literature. AB - The National Asthma Education and Prevention Program recently published updated guidelines that stress the importance of peak flow monitoring for patients with moderate-to-severe persistent asthma. In this specific patient population, a peak flowmeter provides a simple, quantitative, objective measurement of large airway function. The purpose of this article is to describe indications for peak flow monitoring in asthmatic patients, review technical requirements for peak flowmeters as described by the National Heart, Lung, and Blood Institute, and evaluate the literature on commercially available peak flow devices to aid the health professional in selecting an appropriate meter for the patient with moderate-to-severe persistent asthma. PMID- 9726752 TI - Problems encountered in high-level research in developing countries. PMID- 9726750 TI - Perceptions of prognosis, treatment, and treatment impact on prognosis in non small cell lung cancer. AB - STUDY OBJECTIVES: We designed and performed a survey to assess the roles and knowledge level of physicians, by specialty, in the management of non-small cell lung cancer (NSCLC). The primary objective was to evaluate the potential variability of physician perceptions of prognosis, recommended treatments, and the impact of treatment on prognosis for each stage of NSCLC. DESIGN: A random national sample of physicians from one of five specialties (primary care, pulmonary medicine, medical oncology, radiation oncology, or thoracic surgery) were invited to complete a 15-min questionnaire. Five case scenarios were presented, each representing a different stage of NSCLC. Participants were asked to state the expected prognosis, recommended treatment, and expected impact of the recommended treatment on prognosis for each scenario. MEASUREMENTS AND RESULTS: Of the 442 physicians contacted, 350 completed the survey (86% response rate). In general, physicians agreed that the primary treatment of patients with stage I disease should be surgery. For all other stages of disease, however, there was a wide range of opinion regarding the treatment of choice and expected impact of treatment on prognosis. In particular, a significant proportion of physicians recommended only supportive care for patients with stage IV disease, despite demonstrated benefits of chemotherapy. CONCLUSIONS: Physicians from differing specialties have varied opinions on the management of NSCLC, particularly stages II to IV. Education regarding state-of-the-art therapy should be directed not only at traditional audiences (ie, medical oncologists) but also the major physician specialties involved in the care of NSCLC patients. Wider implementation of treatment advances in late-stage disease could significantly increase the numbers of patients living for a longer time. PMID- 9726751 TI - The timing of tracheotomy: a systematic review. AB - STUDY OBJECTIVE: To examine the impact of the timing of tracheotomy on the duration of mechanical ventilation, the secondary changes to the trachea, and the clinical course of critically ill patients in the ICU. DESIGN: A systematic review of the literature. METHODS: Two independent reviewers conducted a MEDLINE search for relevant literature in the form of randomized or observational controlled clinical studies. Studies were selected for review by criteria determined a priori; and the methodologic quality of selected studies was evaluated by duplicate independent review, also using criteria determined a priori. RESULTS: Five studies were identified, of which three were quasirandomized and none were blinded. Agreement between reviewers of methodologic quality was high (kappa=0.87). CONCLUSIONS: There is insufficient evidence to support that the timing of tracheotomy alters the duration of mechanical ventilation or extent of airway injury in critically ill patients. PMID- 9726753 TI - Mediastinoscopic subtotal removal of mediastinal cysts. AB - STUDY OBJECTIVE: Evaluate the use of mediastinoscopy in the surgical diagnosis and treatment of mediastinal cystic masses in adults. DESIGN: Case reports and literature review. SETTING: Academic department of surgery. PATIENTS: Three consecutive adults with mediastinal masses identified on plain radiographs and CT. INTERVENTIONS: Operative mediastinoscopy. MEASUREMENTS AND RESULTS: All patients were successfully treated with removal of cyst wall, establishment of diagnosis, and same-day hospital discharge. CONCLUSIONS: Simple mediastinoscopic removal of mediastinal cysts offers the same potential for diagnosis and treatment as more conventional methods, with a potential for less morbid and more cost-effective care. PMID- 9726754 TI - Dyspnea and chest pain associated with lung mass. PMID- 9726755 TI - Acute mental changes in a 68-year-old man with bladder cancer. PMID- 9726756 TI - Bilateral pneumothorax following air bag deployment. AB - Air bags have been shown to decrease mortality from automobile accidents. Herein is a unique case of bilateral pneumothorax following deployment and rupture of an air bag with no other associated chest trauma. One may posit that rupture of the air bag allowed high-pressure gases to be expelled into the patient's lungs resulting in explosive barotrauma. PMID- 9726757 TI - A case of concomitant tuberculosis and sarcoidosis with mycobacterial DNA present in the sarcoid lesion. AB - A 35-year-old Chinese woman initially presented with histologically and bacteriologically confirmed tuberculous lymphadenitis. She was also found to have thrombocytopenia, elevated serum alkaline phosphatase, and bilateral lung infiltrates. After 15 months of antituberculosis treatment, despite resolution of the cervical lymphadenopathy, she started to experience dyspnea. Chest radiograph appearance, thrombocyte count, and liver biochemistry had all deteriorated as well. Histologic findings from tissues obtained via transbronchial biopsy and open lung biopsy were consistent with sarcoidosis but also showed the presence of mycobacterial DNA by the polymerase chain reaction. She subsequently achieved a very good response clinically, radiographically, hematologically, and biochemically with 1-year of corticosteroid treatment for her sarcoidosis, and she remained relapse-free afterwards. The concomitant presence of tuberculosis and sarcoidosis in this patient together with the presence of mycobacterial DNA in the sarcoid lesion reiterate the possibility that mycobacteria or some of its components may be capable of inducing the immune response and the pathologic changes of sarcoidosis. PMID- 9726759 TI - Enuresis and obstructive sleep apnea in adults. AB - Adult enuresis is an unusual symptom of obstructive sleep apnea (OSA). Although it is described as a classic symptom of childhood OSA, enuresis is encountered infrequently in adult sleep medicine. Five adults with enuresis associated with sleep apnea presented to our Sleep Disorders Center. In all five cases, the onset of enuresis was associated with the progression of sleep apnea symptoms. In each case, the enuresis resolved with treatment with nasal continuous positive airway pressure. Current medical literature on the postulated mechanisms of nocturia and enuresis in sleep apnea is reviewed. Based on the experience of the authors and review of the medical literature, one may conclude that severe OSA may lead to new-onset enuresis in adults and that effective treatment of OSA is associated with resolution of enuresis. PMID- 9726758 TI - Acute community-acquired pneumonia due to Aspergillus in presumably immunocompetent hosts: clues for recognition of a rare but fatal disease. AB - This article reports a case of acute community-acquired pneumonia due to Aspergillus fumigatus in a healthy patient and reviews 11 previously reported cases occurring in presumably immunocompetent hosts. The diagnosis was delayed for all patients; mortality was 100%. Clues that might suggest Aspergillus as a pathogen in community-acquired pneumonia include a chest radiograph revealing diffuse infiltrates or new cavitation; lack of bacterial or viral cause; a preceding influenza A infection; and respiratory secretion cultures positive for Aspergillus. When these clues are present, the physician should consider an early biopsy of lung tissue. Increased recognition and more timely diagnosis in future cases will improve the outcome of this rare but fatal infection. PMID- 9726760 TI - Migration and right atrial perforation of an Accufix atrial lead retention wire following partial lead removal during myomectomy. AB - A 36-year-old man with a history of hypertrophic obstructive cardiomyopathy presented to the emergency room with "stabbing" chest pain. He had undergone dual chamber pacemaker implantation in 1993 using an atrial lead (Accufix; Telectronics; Englewood, Colo) and a myomectomy in 1996 during which the distal portion of the atrial lead was removed. Digital fluoroscopy revealed that the retention wire had migrated out of the remaining atrial lead and perforated the right atrium. The retention wire was successfully removed percutaneously. The need for complete removal of the retention wire in the Accufix lead at the time of open-heart surgery is emphasized. PMID- 9726761 TI - Metastatic breast disease 40 years after the initial diagnosis. AB - This is the case of a patient with metastatic disease diagnosed 40 years after a radical mastectomy which was followed by radiation treatment for breast cancer. The patient had nonspecific symptoms for 3 years, and a lymph node biopsy revealed the underlying cause to be recurrent breast cancer. Excision of the largest metastases combined with chemotherapy resulted in a further 3-year remission. PMID- 9726762 TI - Gemfibrozil-warfarin drug interaction resulting in profound hypoprothrombinemia. AB - The following describes a patient on a stable regimen of warfarin who developed severe hypoprothrombinemia and bleeding 4 weeks after starting gemfibrozil. Despite a warning by the manufacturer, only one report of this interaction has been published in the literature. This interaction may be overlooked by clinicians, which may result in a serious bleeding risk for patients on warfarin. PMID- 9726763 TI - Pyogenic vertebral osteomyelitis presenting as exudative pleural effusion: a series of five cases. AB - Five patients are reported who have pleural effusion and pyogenic vertebral osteomyelitis. In four of the five patients, the presenting problem was a large pleural effusion, and three of these four patients had an exudative effusion. Initial evaluation and investigations in these patients were directed toward the pleuropulmonary disease, delaying the diagnosis of osteomyelitis, and in two patients, this delay resulted in neurologic complications. In the fifth patient, the pleural effusion was initially small; however, during the course of a workup for osteomyelitis, the effusion increased rapidly. Two out of the five patients had empyema, and the other three patients had a large pleural effusion associated with and apparently caused by vertebral osteomyelitis. Vertebral osteomyelitis should be considered in the differential diagnosis of pleural effusion of uncertain cause especially if there is associated back pain. PMID- 9726764 TI - Primary cardiac sarcoma: a novel treatment approach. AB - Primary cardiac sarcomas carry a dismal prognosis with no known curative therapy using standard treatment approaches. By its very location, the possibility of a radical complete resection--the underlying principle in the management of any soft-tissue sarcoma--is precluded. While literally in a continuous "blood bath," cardiac sarcomas are associated with a very high rate of hematogenous metastases. This report describes the management of a case in a 51-year-old white man with a high-grade unresectable cardiac sarcoma who was treated with hyperfractionated (twice daily) radiotherapy to a total dose of 7,050 cGy along with a radiosensitizer, (5'-iododeoxyuridine. The patient currently is disease-free and functioning well more than 5 years following this novel treatment approach. PMID- 9726765 TI - Code 99--who's watching? PMID- 9726766 TI - Chopin's illness. PMID- 9726767 TI - The long suffering of Frederic Chopin, revisited. PMID- 9726769 TI - Use of nonheparinized syringes for collecting pleural fluid samples. PMID- 9726768 TI - Chopin's malady. PMID- 9726771 TI - Significance of airway colonization by Burkholderia gladioli in lung transplant candidates. PMID- 9726770 TI - Nonprescription bronchodilator use in asthma. PMID- 9726772 TI - Parapneumonic effusion does not equal empyema. PMID- 9726773 TI - Esophageal disorders and chronic cough in children. PMID- 9726774 TI - Nasal bridge oximetry: an alternative site in poor peripheral pulsations. PMID- 9726775 TI - Lung cancer staging--a proposal. PMID- 9726776 TI - Thoracic tumors in pregnant women. PMID- 9726777 TI - Effects of motorcycle exhaust on cytochrome P-450-dependent monooxygenases and glutathione S-transferase in rat tissues. AB - The effects of motorcycle exhaust (ME) on cytochrome P-450 (P-450)-dependent monooxygenases were determined using rats exposed to the exhaust by either inhalation, intratracheal, or intraperitoneal administration. A 4-wk ME inhalation significantly increased benzo[a]pyrene hydroxylation, 7 ethoxyresorufin O-deethylation, and NADPH-cytochrome c reductase activities in liver, kidney, and lung microsomes. Intratracheal instillation of organic extracts of ME particulate (MEP) caused a dose- and time-dependent significant increase of monooxygenase activity. Intratracheal treatment with 0.1 g MEP extract/kg markedly elevated benzo[a]pyrene hydroxylation and 7-ethoxyresorufin O deethylation activities in the rat tissues 24 h following treatment. Intraperitoneal treatment with 0.5 g MEP extract/kg/d for 4 d resulted in significant increases of P-450 and cytochrome b5 contents and NADPH-cytochrome c reductase activity in liver microsomes. The intraperitoneal treatment also markedly increased monooxygenases activities toward methoxyresorufin, aniline, benzphetamine, and erythromycin in liver and benzo[a]pyrene and 7-ethoxyresorufin in liver, kidney, and lung. Immunoblotting analyses of microsomal proteins using a mouse monoclonal antibody (Mab) 1-12-3 against rat P-450 1A1 revealed that ME inhalation, MEP intratracheal, or MEP intraperitoneal treatment increased a P-450 1A protein in the hepatic and extrahepatic tissues. Protein blots analyzed using antibodies to P-450 enzymes showed that MEP intraperitoneal treatment caused increases of P-450 2B, 2E, and 3A subfamily proteins in the liver. The ME inhalation, MEP intratracheal, or MEP intraperitoneal treatment resulted in significant increases in glutathione S-transferase activity in liver cytosols. The present study shows that ME and MEP extract contain substances that can induce multiple forms of P-450 and glutathione S-transferase activity in the rat. PMID- 9726778 TI - Analysis of air pollution particulate-mediated oxidant stress in alveolar macrophages. AB - Adverse health effects of urban air pollution particulates may be attributable to particle-mediated oxidant stress and inflammation. Intracellular oxidant production in normal hamster alveolar macrophages (AMs) was measured upon exposure to concentrated ambient particulates (CAPs), residual oil fly ash (ROFA), and their water-soluble and particulate fractions. ROFA and CAPs caused increases in dichlorofluorescin (DCFH) oxidation, a fluorescent measure of intracellular reactive oxygen species (ROS) production, comparable to the positive control, phorbol myristate acetate (PMA). The water-soluble component of both CAPs and ROFA (CAPs, S and ROFA, S) significantly increased AM oxidant production over negative control. CAPs samples and components showed substantial day-to-day variability in their oxidant effects. Metal chelation by desferrioxamine (DF, 1 mM) caused significant inhibition of particulate-induced AM oxidant production. ROFA exposure resulted in increased macrophage inflammatory protein-2 (MIP-2) message in AMs and in increased tumor necrosis factor alpha (TNF-alpha) production by the monocyte-macrophage cell line, RAW 264.7. TNF-alpha production was inhibitable by the antioxidant N-acetylcysteine (NAC). The data suggest that metal components adsorbed to urban air pollution particulates can significantly contribute to particulate ability to cause oxidant stress and cytokine production in AMs. PMID- 9726779 TI - Acute exposure to mercury from amalgam: no short-time effect on the peripheral blood lymphocytes in healthy individuals. AB - Mercury, released from dental amalgam, has been considered to adversely affect the human immune system. This study has been performed in order to evaluate if an acute low-dose mercury exposure, achieved by total amalgam removal in 10 healthy individuals, would affect the immunocompetent cells in human blood when the mercury level in blood and plasma was increasing. Induction of lymphocyte proliferation, measured as spontaneous de novo DNA synthesis, and total T cells, CD4+ T cells, CD8+ T cells, and B cells, was studied prior to and 7, 31, and 48 h after amalgam removal. In addition, the levels of interleukin-6 (IL-6) and C reactive protein (CRP) in serum/plasma were measured. Despite a significant increase of the plasma mercury levels within 24 h after intervention, no significant influence on the peripheral blood lymphocytes could be detected during the first 48 h. The serum IL-6 levels increased significantly within 48 h after intervention, but were still low and within normal range. No influence on the CRP levels up to 7 d after amalgam removal was detected. PMID- 9726780 TI - Lack of suppressive effects of mixtures containing low levels of methylmercury (MeHg), polychlorinated dibenzo-p-dioxins (PCDDS), polychlorinated dibenzofurans (PCDFS), and aroclor biphenyls (PCBS) on mixed lymphocyte reaction, phagocytic, and natural killer cell activities of rat leukocytes in vitro. AB - Rat splenocyte mixed leukocyte reaction (MLR), splenic natural killer (NK) cell activity, and phagocytic activities of splenic, peritoneal, and peripheral blood leukocytes (PBLs) were evaluated in vitro to determine the immunotoxicity of mixtures containing low levels of methylmercury (MeHg), polychlorinated dibenzo-p dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), and Aroclor polychlorinated biphenyls (PCBs). The mixtures were based on the concentrations of the chemicals in fish flesh. Leukocytes from male Fischer rats were exposed to MeHg (0.1-2 microg/ml), PCDD/PCDF mixtures (1-15 pg/ml) of three PCDDs (2,3,7,8 tetrachlorodibenzo-p-dioxin, 1,2,3,7,8-pentachlorodibenzo-p-dioxin, and 1,2,3,4,7,8-hexachlorodibenzo-p-dioxin) and two PCDFs (2,3,7,8 tetrachlorodibenzofuran and 1,2,3,7,8-pentachlorodibenzofuran), three Aroclor PCB (Aroclor 1242, 1254, and 1260) mixtures (0.01-0.5 microg/ml), or combinations of MeHg/PCB/PCDD/PCDF mixtures for 24 or 72 h before immunological assays. Phagocytosis and NK cell cytotoxicity were evaluated with a flow cytometer, and MLR of Fischer rat responder splenocytes cultured with mitomycin C-treated Long Evans splenocytes by [3H]thymidine uptake. Exposure to MeHg (2 microg/ml) alone or with PCB/ PCDD/PCDF resulted in significant cytolethality in rat splenocytes, peritoneal leukocytes, and PBLs at 24 h exposure. Treatment with Aroclor PCB mixtures, PCDD/PCDF mixtures, 0.1 microg MeHg/ml (noncytolethal), or PCB/PCDD/PCDF mixtures with 0.1 microg MeHg/ml caused no suppression of splenocyte MLR response, splenic NK cell-mediated lysis of Yac-l cells, or phagocytosis of fluorescent beads by splenic, peritoneal, and peripheral blood phagocytic cells. The results indicate that in vitro exposure of rat leukocytes to low levels of MeHg, Aroclor PCB mixtures, PCDD/PCDF mixtures, or MeHg/PCB/PCDD/PCDF mixtures had no suppressive effects on the immune functions assayed, and thus produced no additive immunotoxicity. However, in order to predict the potential risk of these chemical mixtures to the human immune system, in vivo animal studies with blood (tissue) levels compatible with the levels of MeHg, PCBs, and PCDDs/PCDFs in exposed human populations should be evaluated. PMID- 9726781 TI - Environmental causes for sinonasal cancers in pet dogs, and their usefulness as sentinels of indoor cancer risk. AB - A case-control study was conducted to investigate the environmental causes of sinonasal cancers among pet dogs. Sinonasal cancer (SNC) cases and digestive cancer controls from the years 1989 through 1993 were obtained from a veterinary histopathology database. Owners were mailed a self-administered survey requesting information on canine factors, owner demographics, household exposures (including environmental tobacco smoke), and local pollution. A total of 129 case owners and 176 control owners returned completed surveys: a response rate of approximately 72%. Only household exposures were associated with increased SNC risk. Use of indoor coal or kerosene heaters represented the strongest risk factors, with significant adjusted odds ratios of 4.2 and 2.2 respectively. Environmental tobacco smoke exposure was not a risk factor and was suggestive of a nonsignificant, mildly protective effect at the lower exposure levels. Increasing nasal length was a significant risk factor, and there was effect modification between nasal length and coal or kerosene combustion. No self-reported measures of local pollution, such as urban status or residence within 1 mile of a factory, were associated with SNC risk. These results suggest that canine SNC has a strong environmental component and highlight the importance of indoor exposures, especially from fossil fuel combustion products. These results also suggest that pet dogs represent excellent sentinels for indoor cancer risk and that canine SNC cases can be used as early markers of household exposure to carcinogenic combustion products. PMID- 9726782 TI - Heavy metals and fertility. AB - Heavy metals have been identified as factors affecting human fertility. This study was designed to investigate whether the urinary heavy metal excretion is associated with different factors of infertility. The urinary heavy metal excretion was determined in 501 infertile women after oral administration of the chelating agent 2,3-dimercaptopropane-1-sulfonic acid (DMPS). Furthermore, the influence of trace element and vitamin administration on metal excretion was investigated. Significant correlations were found between different heavy metals and clinical parameters (age, body mass index, nationality) as well as gynecological conditions (uterine fibroids, miscarriages, hormonal disorders). Diagnosis and reduction of an increased heavy metal body load improved the spontaneous conception chances of infertile women. The DMPS test was a useful and complementary diagnostic method. Adequate treatment provides successful alternatives to conventional hormonal therapy. PMID- 9726783 TI - Gender differences in acute N-(3,5-dichlorophenyl)-2-hydroxysuccinimide (NDHS) and N-(3,5-dichlorophenyl)-2-hydroxysuccinamic acid (2-NDHSA) nephrotoxicity in Fischer 344 rats. AB - N-(3,5-Dichlorophenyl)succinimide (NDPS) is an agricultural fungicide that induces nephrotoxicity as its major toxicity. NDPS is also a more potent nephrotoxicant in female than in male rats. The purpose of this study was to examine the nephrotoxic potential of the two NDPS metabolites N-(3,5 dichlorophenyl)-2-hydroxysuccinimide (NDHS) and N-(3,5-dichlorophenyl)-2 hydroxysuccinamic acid (2-NDHSA) in age-matched male and female Fischer 344 rats to determine if gender differences exist for the nephrotoxicity induced by the two NDPS metabolites. Rats (4 per group) were administered a single intraperitoneal (ip) injection of NDHS or 2-NDHSA (0.025 or 0.05 mmol/kg) or vehicle, and renal function was monitored for 48 h. Neither compound induced significant nephrotoxicity in male rats at the doses tested. However, in female rats both metabolites induced marked nephrotoxicity at the 0.05 mmol/kg dose level, and treatment with 0.025 mmol/kg 2-NDHSA induced some changes in renal function (transient diuresis, transient proteinuria, decreased organic ion accumulation). Little effect on renal function was induced in females by treatment with 0.025 mmol/kg NDHS. At toxic levels in female rats, the renal lesions were located primarily in the S2 and S3 segments of the proximal tubule. These results indicate that, like the parent compound, gender differences exist in the nephrotoxic potential of NDHS and 2-NDHSA. The results also suggest that in females, as in males, NDPS nephrotoxicity is mediated via NDHS and/or 2-NDHSA. However, it is not clear if the ultimate nephrotoxicant species following NDPS exposure is different in males and females or if the same ultimate nephrotoxicant species is produced in both species but handled differently by male and female kidneys. Thus, further studies are needed to determine the exact nature of the ultimate nephrotoxicant species and the mechanisms of the observed gender differences. PMID- 9726784 TI - Inhibition of rat lung mixed-function oxidase activity following repeated low level toluene inhalation: possible role of toluene metabolites. AB - Toluene is a commonly used solvent that has been shown to alter mixed-function oxidase (MFO) activity, in an organ- and isozyme-specific pattern, following intraperitoneal administration. The purpose of this study was to determine whether similar changes occurred following repeated, low-level inhalation exposure, and to investigate the role of toluene metabolites in these alterations. Exposure to 375 ppm toluene, 6 h/d for up to 5 d, resulted in significant inhibition of the activity of pulmonary arylhydrocarbon hydroxylase (AHH), cytochrome P-4502B1 (CYP2B1), and CYP4B1, but not CYP1A1. After exposure to lower toluene levels (125 ppm, 6 h/d, 3 d), the activities of lung AHH, CYP2B1, and CYP4B1 were also significantly decreased, but in a dose-related manner. MFO activity was not consistently altered in liver. Control pulmonary or liver microsomes were incubated with various concentrations (0.01-10 mM) of toluene or its metabolites and CYP2B1, CYP1A1, and/or CYP4B1 activities were subsequently determined. Benzaldehyde produced a significant dose-related inhibition in the activity of all three lung P-450s examined (IC50 10(-3) M). Toluene was found to be a more potent inhibitor of lung CYP2B1 and CYP1A1 (IC50, 10(-4) M) than benzaldehyde, but neither toluene nor benzyl alcohol was an effective inhibitor of lung CYP4B1. Toluene and its metabolites were weaker inhibitors of CYP1A1 than of CYP2B1. For CYP2B1 and CYP1A1, the order of inhibitory potency was toluene > benzaldehyde > benzyl alcohol and suggests that both the parent molecule and its metabolites may act in concert to inhibit catalytic activity of these cytochromes. The MFO inhibition seen after repeated low-level toluene inhalation exposure could result in altered metabolic profiles of other xenobiotics in an organ-specific fashion. PMID- 9726785 TI - Thermoregulation in rats exposed perinatally to dioxin: core temperature stability to altered ambient temperature, behavioral thermoregulation, and febrile response to lipopolysaccharide. AB - Recent studies have shown that perinatal exposure to 2,3,7,8-tetrachlorodibenzo-p dioxin (TCDD, dioxin) alters thermoregulatory function in adult rats and hamsters, indicated by a reduced body temperature during the animal's nocturnal phase. The present study was designed to assess the behavioral thermoregulation, ability to develop a fever, and thermoregulatory stability as a function of ambient temperature (Ta) in rats exposed perinatally to TCDD. Pregnant Long-Evans rats were exposed on gestational day (GD) 15 to 1 microg TCDD/kg (po). The male offspring were implanted with transmitters to monitor core temperature (Tc) and motor activity (MA). The 24-h pattern of core temperature was affected by TCDD exposure, characterized by a reduced nocturnal Tc. At some ages, the diurnal Tc of the TCDD group was elevated. This dysfunction in temperature regulation was most apparent at 7 and 11 mo of age. The 24-h pattern of MA was also altered by TCDD. The hypothermic effects of TCDD were most pronounced at cooler Ta values of 10 to 22 degrees C. In contrast, behavioral thermoregulation, assessed by measuring the selected Ta and Tc of rats in a temperature gradient, was unaffected by TCDD. The ability to develop a fever following administration of lipopolysaccharide (LPS) endotoxin (Escherichia coli; 50 microg/kg) was accentuated in the TCDD-treated animals. The data confirm a nocturnal hypothermia in rats prenatally exposed to TCDD. However, the normal behavioral regulation of Tc suggests that hypothalamic thermoregulatory centers are not permanently altered. The accentuated fever in TCDD animals shows possible functional alterations in the neuroimmune and/or thermoregulatory axes involved in fever. PMID- 9726786 TI - Selenium research in Serbia, Yugoslavia. AB - Data on selenium (Se) deficiency in Serbia are presented following 10 years of research. We studied the Se content in ores, stream sediment, soil, cereal crops, and garlic grown in these soils, food, and human serum and hair from 55 communities. Most of the results indicated a serious Se deficiency. In some communities, the Se content of grain, garlic, and human serum and hair approached that of the low-Se belt in China. We assume that an extremely low Se level in the human population could be a risk factor for the development of Balkan nephropathy (BN) and for the very high incidence of urinary tract tumors (UTT) in endemic areas, as well as high mortality rates of malignant diseases. On the other hand, some regions with relatively higher serum Se levels had significantly lower mortality rates of cancer and cardiovascular diseases. PMID- 9726787 TI - Geochemistry of selenium. AB - Selenium (Se) is one of the most peculiar chemical elements in the geo- and biospheres. It partly resembles sulfur and tellurium; however, its behavior in the geosphere and its functions in the biosphere are very specific. Despite a relatively large database, its cycling in both the natural environment and in that modified by human activities requires further study. Selenium is rather concentrated in the geospheric cycle and is also bioconcentrated. The values of its accumulation ratios are: 5 for soil/sandstone, 2 for animal tissues/sandstone, and 5 for animal tissues/grain. For a specific plant/soil system, the bioconcentration factor for plants always has to be estimated because some plants can absorb extremely high concentrations of Se. Their ability to accumulate and tolerate high Se levels is related to different Se metabolisms. These plants play a significant role in geochemical prospecting and animal nutrition. This paper presents some geochemical observations toward a better understanding of the environmental properties of Se. PMID- 9726788 TI - Selenium status in soils in Yugoslavia. AB - In the last 8 years, a number of data on selenium (Se) content and distribution in the soil of Yugoslavia (Serbia and Montenegro) became available. The Se content of the soil in Yugoslavia varies in a broad range 39 to 440 microg/kg (mean value 230 microg/kg; n = 284). The soil clay fraction is rich in Se (range 146 to 586 microg/kg) in relation to sand and silt fractions. The available Se content (after extraction with ammonium acetate and EDTA) varies in the range of 1.2 to 28.2% of the total Se content. The speciation of Se is shown for the soils derived from volcanic rocks in Serbia. In addition, the influence of some soil properties on the Se content, as well as its content in the rocks, sediment, and wild plants in Yugoslavia, is discussed. The geographical distribution of Se in the soil of Yugoslavia shows that it is found in inadequate amounts in many agricultural regions. Its low content in soil has been thought to be associated with a higher incidence of some diseases. The Se content of the soil in Yugoslavia is not fully known. There is a great need to make a systematic geochemical mapping for Se and other trace elements in the soil. PMID- 9726789 TI - Influence of long-term fertilization on the selenium content of calcareous chernozem soil. AB - Available data on the selenium (Se) content in Yugoslavian soils indicate that the element is present in small amounts (< 500 microg/kg in Se-deficient soils). There are no data on the effect of various fertilizers on the Se content in Yugoslavian soils. In our study, we examined the effect of the long-term application of mineral and organic fertilizers (farmyard manure and cornstalks) on the content of Se in calcareous chernozem soil. The experiment of the Maize Research Institute in Belgrade was set up in 1971 and soil samples were examined by chemical analysis after 23 years. The following important conclusions can be drawn based on the analytical data obtained. The total content of Se in the investigated experimental variants in calcareous chernozem soil ranges from 166 to 593 microg/kg. All the variants had a higher content of Se than soil samples taken before the experiment was set up. Comparison with the control (variant without fertilizers) indicated that the Se content increased in the experimental variants where farmyard manure had been applied. This effect was noticed to a depth of 80 cm. Application of farmyard manure should be considered as a means of increasing the levels of Se in Se-deficient soil. Correlation coefficients between total Se content in the soil and some important agrochemical properties of the investigated soils are presented in this paper. PMID- 9726790 TI - Selenium content in sulfide ores from the Chalkidiki peninsula, Greece. AB - Selenium (Se) was assessed in galena, sphalerite, and pyrite samples. These are components of mixed sulfide ores from the Olympias and Madem Lakkos-Mavres Petres deposits and the Skouries porphyry-copper deposit. We used atomic absorption spectroscopy (AAS) with a hydride generator system. The highest concentration of Se (516 ppm) was found in the fine-grained galena at the -135 level of the Olympias deposits. In the Madem Lakkos-Mavres Petres deposit, the highest concentration of Se (33 ppm) was found in the pyrites of the level 30. The concentration of Se in the arsenopyrites and chalcopyrites is lower than the detection limit of the analytical method (< 100 ppb). The concentrated chalcopyrite from the porphyry copper deposit at Skouries exhibits a significant Se content (average 200 ppm) in contrast to the chalcopyrite from the Olympias and the Madem Lakkos-Mavres Petres. Variations in the Se content of the sulfide minerals studied could be caused by redox-pH and/or temperature conditions, as well as by the difference in crystal structure. The Se found in the areas studied may positively affect the environment. Sulfide minerals are oxidized by microorganisms, infiltrate in the soil-water in the form of selenate or selenite ion, and directly or indirectly influence the human organism. PMID- 9726791 TI - Selenium content of sulfide ores related to ophiolites of Greece. AB - Several deposits of sulfide mineralization have been described in the ophiolites of Greece. Based on their mineralogical and chemical composition and the host rocks, two types can be distinguished: (1) the Fe-Cu-Ni-Co type consisting of pyrrhotite, chalcopyrite, Co-pentlandite, pyrite, magnetite + arsenides, +/- chromite, hosted in serpentinites, gabbros or diabases, which have variable geochemical characteristics, and (2) sulfide mineralization of the Cyprus type containing variable proportions of pyrite, chalcopyrite, bornite, and sphalerite. The spatial association with shear zones and fault systems, which is a common feature in both types of mineralization, provided the necessary permeability for the circulation of the responsible mineralized hydrothermal fluids. The selenium (Se) content in representative samples of both types of mineralization from the ophiolites of Pindos (Kondro, Perivoli, and Neropriona), Othrys (Eretria and A. Theodoroi), Veria (Trilofon), and Argolis (Ermioni) shows a wide variation. The highest values of Se (130 to 1900 ppm) were found in massive Fe-Cu sulfide ores from Kondro, in particular the Cu-rich portions (average 1300 ppm Se). The average values of Se for the Othrys sulfides are low (< 40 ppm Se). The Se content in a diabase breccia pipe (50 x 200 m) with disseminated pyrite mineralization (Neropriona) ranges from < 1 to 35 ppm Se. The highest values were noted in strongly altered samples that also exhibited a significant enrichment in platinum (1 ppm Pt). Sulfide mineralization (irregular to lens-like masses and stringers) associated with magnetite, hosted in gabbros exposed in the Perivoli area (Tsouma hill), shows a content ranging from 40 to 350 ppm Se. The distribution of Se in the studied type of the sulfide mineralization may be of genetic significance, indicating that the Se level, which often is much higher than in typical magmatic sulfides related to mafic-ultramafic rocks (average 90 100 ppm Se), may positively affect the environment. PMID- 9726792 TI - Selenium and arsenic in the environment in Finland. AB - A characteristic feature of glaciated Precambrian environments is their low selenium content, as a chalcophile element, Se, replaces sulfur in many of the sulfide minerals, for example, pyrite, chalcopyrite, pyrrhotite, and pentlandite. The average Se concentration in rocks and related till deposits in Finland is in the range of 0.01 to 0.2 mg/kg. Due to geological conditions, Se concentrations in surface and ground water are low in Finland compared with other countries. In a nationwide study dealing with the hydrogeochemistry of headwater streams, the median Se concentration in streams during August to September 1990 was 30 to 180 microg/L. For comparison, Se concentrations in shallow well waters are generally in the range of 50 to 1000 microg/L. The Se concentrations in stream sediments varied from 0.03 to 3.94 mg/kg. There was a highly significant correlation between the Se concentrations in stream water and in stream sediment. The streams with Se concentrations exceeding the general level in both water and sediment were most common in southern Finland. A speciation study on Finnish stream waters revealed that there were equal proportions of Se complexed with humic substances (36%) and Se as a selenate species (36%), whereas selenite accounted for less than 10% of total Se. About 8% of the Se in stream water occurred in particulate form. In an effort to enhance the Se intake of Finns through diet, Se supplemented fertilizers have been used nationwide since 1985. While greatly improving Se levels in the population, the measure has raised concerns about undesirable environmental effects. Therefore, the amount of Se added to fertilizers has been reduced since 1991. Differing in behavior from Se, arsenic is considered one of the most toxic metals derived from the natural environment. Alarm has been triggered in Finland by the recent lowering from 50 microg/L to 10 microg/L of the upper level of As permissible in potable water, the recent information of high As concentrations in water from drilled bedrock wells, and the findings of international medical studies suggesting that As is a carcinogen. The most important source of As is arsenopyrite (FeAsS). Hence, high As concentrations most frequently occur in areas of sulfide mineralization, often in connection with occurrences of mafic rocks such as gabbros, amphibolites, and peridotites. The As concentrations in till fines, the most common glaciogenic soil type in Finland, reflect those in bedrock. The concentrations in groundwater are controlled by the chemical composition of the bedrock and the soil and prevailing hydrogeochemical conditions, for example, pH and Eh levels. Arsenic concentrations are lowest in surface water and swiftly flowing shallow ground water discharged by springs and are somewhat higher in shallow wells dug into overburden. By far, the highest As concentrations are to be found in wells drilled into bedrock (maximum 1 to 2 mg/L), although the concentrations vary by several orders of magnitude from well to well. The highest probability of encountering deleteriously arsenious well water is in areas with characteristic As anomalies in the till and bedrock. Hence, it is important to understand local geological conditions, particularly in the case of wells drilled into bedrock. The risk of deleteriously high As concentrations occurring in captured springs and shallow wells is slight. PMID- 9726793 TI - Variation of selenium content in growing wild plants during vegetative period. AB - There is evidence that selenium (Se) is essential for humans, animals, and some species of microorganisms. This study examines the dynamics of the change of Se accumulation in some growing wild plants: Astragalus onobrychis var. Chlorocarpus (Gris.) Stoj. et Stef. (Leguminosae), Oxytropis pilosa (L.) DC., Salvia officinalis, L., and Simphytum ottomanum L. The influences of some ecological factors (Se content in soil, temperature, and rainfall) have also been surveyed. The Se content of plants from Serbia was 44.5 to 177.0 microg/kg of dry weight. Astragalus onobrychis var. Chlorocarpus (Gris.) Stoj. et Stef. (Leguminosae) accumulates the highest Se content in the blooming stage (161.3 microg/kg). All soils studied were deficient in selenium (< 250 microg/kg). We found a significant correlation between Se content in plants and in the soil (p = 0.88). Rainfall has an influence on Se content in Oxytropis, Salvia, and Simphytum, but not in Astragalus. Our results suggest that Se cannot have a similar role in all plants examined. PMID- 9726794 TI - Selenium in soil, grass, and human serum in the Zlatibor mountain area (Serbia): geomedical aspects. AB - The Zlatibor district in Serbia has lower mortality rates of malignant and cardiovascular diseases (CVD) compared with other regions in Serbia. To better understand the influence of the geochemical environment, we collected and analyzed soil from various bedrocks and the grass growing on them. We also analyzed spring and stream waters, including large water supply accumulations, for major chemical elements and examined the serum of healthy adults in the large area of Zlatibor for selenium (Se) and magnesium (Mg). Our studies included villages, small towns, and the town of Uzice. Our results showed a variable Se content in the soil over different bedrocks. In general, soil in this area has a higher Se content than in other regions of Serbia. The Se content of the grass is influenced by bedrock and soil mineralogy, but mostly by soil pH and the date of collection. For example, in late summer, grass contains twice as much Se than in spring. Mg2+HCO3(-)-type waters occur in the ultramafic massif of Zlatibor in a concentration of 44 to 68 mgMg/L. The serum Se values were higher in the Zlatibor area than in other regions of Serbia (62.6 +/- 14.9 microgSe/L; n = 158). The serum Mg content (22.7 +/- 2.2 mg/L; n = 158) was in the uppermost part of the reference range. Taking into account their biological role, the Se and Mg levels in the human population in the Zlatibor area could influence the lower mortality rates of cancer and CVD in this region compared with other regions in Serbia. PMID- 9726795 TI - Geomedical aspects of selenium: Norwegian investigations. AB - Selenium has many health effects, both beneficial and harmful. Chemical climatology has been a valuable background for explanation of the geomedical effects of this element. Geomedicine is the science dealing with the influence of ordinary environmental factors on the geographical distribution of health problems in humans and animals. PMID- 9726796 TI - Selenium glutathione peroxidase: some aspects in man. AB - The levels of activity of selenium glutathione peroxidase (GSH-Px) in animals, in circulating blood cells, and in pathologic conditions in man are reviewed. The results are discussed in relation to circulating lipoperoxides and to selenium supplementation. PMID- 9726798 TI - Selenium intake as a modulator of responsiveness to oxidative stress. AB - A deficiency in important components of the endogenous antioxidative defense system (AODS) against the production of reactive oxygen species, including free radicals, results in the accumulation of oxidative damage, inducing oxidative stress. A dietary deficiency in selenium (Se), an important part of AODS, can increase the sensitivity of a living system to oxidative stress. We investigated the effects of Se supplementation, in the form of Se-enriched yeast, on the AODS resistance of red blood cells (RBC) to experimentally induced oxidative stress. We analyzed the alterations in main components of the AODS, such as the amount of reduced (GSH) oxidized glutathione (GSSG), Se-dependent glutathione peroxidase (GSH-Px), Se content, catalase (CAT), and thiobarbituric acid reactive substances (TBARS), in RBC of male Wistar rats exposed to gamma rays and supplemented with Se-enriched yeast (SeY) in drinking water. The results suggested that the increased Se level generally exhibited a protective effect against whole body irradiation, reducing the expenditure of the AODS components in defense. These reductions differed depending on the time observed and the parameter investigated but, generally, SeY supplementation induced a faster restoration of the AODS after this kind of oxidative stress. PMID- 9726797 TI - Seasonal changes in the antioxidative defense in ground squirrels (Citellus citellus): possible role of GSH-Px. AB - As seasonal hibernators, ground squirrels decrease their body temperature to 7 degrees C and hibernate during the winter. Maintenance at 30 degrees C prevents seasonal changes of body temperature and animals remain euthermic and active. We measured selenium (Se)-dependent glutathione peroxidase (GSH-Px), as well as the activity of other antioxidative components such as superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), and glutathione-S-transferase (GST) and the amount of low-molecular-weight antioxidants glutathione (GSH), ascorbic acid (AsA), and vitamin E (vit E) in spring, summer, and winter in ground squirrels continuously kept at a temperature of 30 degrees C. We examined liver and interscapular brown adipose tissue (IBAT) as thermogenic tissues, as well as the brain and the kidneys. During the winter, we found a decrease in enzymatic activity and an increase in the level of low molecular antioxidants in all tissues. Correlation analysis revealed a similarity in the composition of antioxidative defense (AD) among the tissues examined. The results obtained clearly demonstrated numerous correlative expressions of antioxidative components in this experimental model, especially of GSH-Px, suggesting the complexity of the system responsible for the maintenance of physiological homeostasis. PMID- 9726799 TI - The effect of cadmium and selenium on the antioxidant enzyme activities in rat heart. AB - Two month-old Wistar male albino rats were exposed during a 30-day period to a daily oral intake ad libitum of either 200 microg/mL Cd (as CdCl2), 0.1 microg/mL Se (as Na-selenite), or the same dosages of Cd + Se in drinking water. The daily intake from the water was calculated to be 15 mg Cd/kg and 7 microg Se/kg. Cadmium (Cd) accumulates in the heart (p < 0.005) and, in rats, decreases both body mass growth (p < 0.005) and heart mass (p < 0.02). Selenium (Se) significantly decreases the negative effect of Cd on body mass growth. In the hearts of Cd-treated rats, cadmium caused the decrease (p < 0.05) of selenium dependent glutathione peroxidase (GSH-Px, EC 1.11.1.9) activity. At the same time, the activities of total superoxide dismutase (total SOD, EC 1.15.1.1), manganese-containing superoxide dismutase (Mn SOD), and copper-zinc-containing superoxide dismutase (CuZn SOD) were increased (p < 0.005). The activities of total SOD, CuZn SOD (p < 0.005), GSH-Px (p < 0.02), and glutathione-S-transferase (GST, p < 0.005) were increased in the hearts of Se-treated rats. However, by concomitant administration of Cd and Se, these changes were diminished (total SOD, GST) or were completely eliminated (Mn SOD, GSH-Px). These results indicate that Se only partly diminishes the effects of Cd cardiotoxicity. PMID- 9726800 TI - The effects of selenium deficiency, dietary selenium, and vitamin E supplementation on the oxidative status of pig liver. AB - The aim of this work was to determine the effect of selenium (Se) deficiency on the porcine liver oxidative stability and to investigate Se content and oxidative status in porcine liver after dietary supplementation with vitamin E (vit E), sodium selenite, and selenized yeast. Experimental animals were fed a basal corn meal, low in Se and vit E, for a 4-week depletion period before being given the experimental diets containing different levels of Se and/or vit E for 5 months. Dietary treatments were the basal diet with no additions (control); the basal diet supplemented with 25 mg of vit E/kg of feed (group I); basal diet + 0.3 mg selenite-Se/kg (group II); basal diet + 0.3 mg selenized yeast-Se/kg (group III); basal diet + 0.1 mg selenite-Se + 10 mg vit E/kg (group IV); and basal diet + 0.3 mg selenite-Se + 25 mg vit E/kg (group V). The Se content in pig liver samples was 33 to 192% lower in the control group than in all the other groups. Dietary Se from selenized yeast had a more pronounced effect on Se level than dietary sodium selenite. The highest Se content was found in liver samples from the Se + vit E supplemented group (group V). All the dietary supplementation schemes significantly improved the oxidative status of porcine liver compared with the control group samples. The best results were obtained by simultaneous dietary supplementation with Se + vit E (groups IV and V) > group III > group II > group I. PMID- 9726801 TI - Seasonal changes in the activity of antioxidative defense in the kidneys of the euthermic ground squirrel (Citellus citellus). AB - The aim of this work was to determine the activity of the antioxidant enzymes: superoxide dismutase (EC 1.15.1.1; SOD), catalase (EC 1.11.1.6; CAT), glutathione peroxidase (EC 1.11.1.9; GSH-Px), glutathione-S-transferase (EC 2.5.1.18; GST), glutathione reductase (EC 1.6.4.2; GR) and the low molecular mass antioxidants: ascorbic acid (ASA) and vitamin E (vit E) in the kidney of ground squirrels during circannual changes. Keeping the ground squirrel at the temperature of thermic neutrality (30 degrees C) provides a stable euthermic state during the whole year and thus any change is due to the circannual rhythm. The highest specific activity of all examined antioxidative defense enzymes in the kidney was found in the spring, when ground squirrels are seasonally the most active. In the summer, lower specific activity of GSH-Px as well as of SOD and CAT were noted and, when expressed per g wet mass, only a decrease in GSH-Px activity was recorded. In the kidney of ground squirrels kept at 30 degrees C, the lowest specific activity of all examined enzymes was found during the winter and, when expressed per g wet mass, only the SOD activity was lower than in the spring and summer. Higher amounts of vitamins C and E were found in the ground squirrel kidneys in the summer. The results obtained in this work demonstrate that circannual regulation of metabolic activity, which is inherent to seasonal hibernators, is also expressed at the level of antioxidative defense in the kidneys. PMID- 9726802 TI - Selenium-dependent GSH-Px in erythrocytes of patients with hypertension treated with ACE inhibitors. AB - This study is aimed at examining whether essential arterial hypertension (HTN) or ACE inhibitors have any effect on erythrocyte selenium (Se)-dependent and Se-non dependent glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) activity. Eleven patients with HTN (2 men and 9 women) and 9 healthy volunteers were included in this study after clinical examination and laboratory investigation. The activities of all three enzymes were determined and then the patients were assigned to receive ACE inhibitor therapy consisting of captopril, 25 to 50 mg daily, or enalapril, 10 to 40 mg daily. After 1 year, the determination of antioxidant enzymes was repeated. Our results showed that the initial values of Se-dependent GSH-Px in patients treated with ACE inhibitors were significantly lower (19.60 +/- 3.50 microM NADPH/min(-1)/mgHb(-1)) compared with the controls (28.64 +/- 4.93 microM NADPH/min(-1)/mgHb(-1); p < 0.001), whereas the activity of Se-non-dependent GSH-Px was significantly enhanced (13.55 +/- 1.46 microM NADPH/min(-1)/mgHb(-1); p < 0.001) compared with the control group (9.44 +/- 0.81 microM NADPH/min(-1)/mgHb(-1); p < 0.001). ACE inhibitors did not significantly change the activity of Se-dependent GSH-Px or Se-non dependent GSH-Px. No significant alteration was observed in SOD activity. PMID- 9726803 TI - Antioxidative defense in human myocardial reperfusion injury. AB - The activity of glutathione peroxidase (GSH-Px) as well as the activities of other antioxidative enzymes such as CuZn superoxide dismutase (CuZn SOD), catalase (CAT), glutathione reductase (GR) in erythrocytes, the plasma activity of glutathione-S-transferase (GST), and the plasma levels of vitamin E and vitamin C were evaluated in nine patients with acute myocardial infarction (AMI). Blood samples were taken before and 1, 3, 6, and 24 hours after the institution of thrombolytic therapy. The results were compared with those in 30 healthy volunteers. A significant decrease in catalase (CAT) activity and vitamin E content in patients before and after thrombolytic therapy as compared with controls was recorded. Our results confirmed that a disturbed oxidative/antioxidative balance is present after AMI and after thrombolytic therapy. PMID- 9726804 TI - Blood and plasma selenium levels and GSH-Px activities in patients with arterial hypertension and chronic heart disease. AB - The purpose of this study was to examine the relationship between arterial hypertension (HTN), chronic heart disease (CHD), and selenium (Se) status. Blood and plasma Se concentrations and Se-dependent GSH-Px activities were determined in 40 patients (HTN = 20; CHD = 20) and 17 healthy volunteers aged 41 to 66 years. Whole blood and plasma Se concentrations were significantly lower in the patients with HTN (19.1% and 26.3%, respectively) and CHD (33.1% and 29.4%, respectively) compared with the values obtained in the controls. The hypertensive patients had lower plasma Se-GSH-Px (26.7%), and those with CHD had both lower whole blood (19.5%) and plasma Se-GSH-Px activities (30.2%). A significant positive correlation between plasma Se-GSH-Px activity and ejection fraction (EF) was found in patients with CHD. There were significant correlations between plasma and whole blood Se concentration, plasma Se concentration and Se-GSH-Px activity, and whole blood Se and Se-GSH-Px activity. Our results showed that hypertensive patients and those with CHD had lower Se levels compared with controls. We conclude that low Se content might be a risk factor for development of HTN and CHD. PMID- 9726805 TI - The efficacy of selenium, WR-2721, and their combination in the prevention of adriamycin-induced cardiotoxicity in rats. AB - It is known that the antineoplastic drug adriamycin (ADR) can cause cardiotoxic effects. Some data imply that pretreatment with selenium (Se) and the radio- and chemoprotector, amifostine (WR-2721), may confer a protective effect. The aim of this study was to evaluate the efficacy of single doses of Se and WR-2721, alone or in combination, in the prevention of acute ADR-induced cardiotoxicity in male Wistar rats. Se, in the form of sodium selenite (1.6 mg/kg i.p.), and WR-2721 (300 mg/kg i.p.) were given 24 hours and 20 minutes, respectively, before ADR (6 mg/kg i.v.). The cardiotoxicity of ADR was recorded 48 hours after its administration because earlier studies revealed that structural damage of the myocardium occurs within this period. Evaluation of these toxic effects, as well as of the cardioprotective efficacy of the administered drugs, was performed using (1) ECG-records before and during the infusion of the proarrhythmogenic compound, aconitine (8 microg/kg/min i.v.) and (2) the serum activity of creatine kinase (CK), aspartate aminotransferase-(AST), lactate dehydrogenase (LDH), and its isoenzyme alpha-hydroxybutyrate dehydrogenase (alpha-HBDH). The results showed that the arrhythmogenic dose of aconitine was significantly reduced in ADR treated rats (57.22 vs. 99.65 microg/kg in control; p < 0.05) and that this proarrhythmogenic compound caused a significant increase in heart rate in such animals compared to controls. Pretreatment with Se, WR-2721, and their combination partly reversed the arrhythmogenic dose of aconitine to control (72.09, 82.1, and 88.99 microg/kg, respectively). Se failed to prevent an aconitine-induced increase in heart rate, whereas WR-2721 and their combination successfully counteracted this effect. In addition, ADR produced a significant increase in the serum activity of all monitored enzymes. Pretreatment with Se failed to prevent this increase, whereas pretreatment with WR-2721 did. The best result was obtained with their combination. We conclude that the radio- and chemoprotector, WR-2721, particularly in combination with Se, may provide a significant protective effect against acute ADR-induced cardiotoxicity in rats. PMID- 9726806 TI - Effect of X-rays on selenium content in rat jaws. AB - It is known that X-radiation of the growing jaws results in impaired dental development. We investigated the role of selenium (Se) in radiation-induced dental growth retardation. The heads of 8-day-old female rats were irradiated with a single dose of 9.6 Gy of X-rays. Another group of animals was irradiated under the same conditions but, for radioprotective purposes, were also in deep hypothermia during radiation exposure. Nonirradiated animals served as controls. The Se content in the upper and lower jaws was analyzed by hydride generation atomic absorption spectrophotometry. In the upper jaws of the animals exposed to irradiation, the Se concentration was significantly lower than in those irradiated under conditions of hypothermia (p < 0.05), although both groups showed no significant difference in Se concentration when compared to nonirradiated controls. In the lower jaws, the concentration of Se was significantly reduced in the irradiated group when compared with controls and the rats irradiated under conditions of hypothermia (p < 0.05). There was no significant difference in Se concentration between rats irradiated under conditions of hypothermia and nonirradiated controls. There was no significant change in Se concentration in the jaws of the rats protected by hypothermia during radiation exposure compared with nonirradiated controls. Although Se loss in the irradiated bone could be prevented by hypothermia, we could not prove that Se loss per se is the cause of growth retardation nor that its retention has a radioprotective effect. PMID- 9726807 TI - Effect of selenium-enriched yeast pretreatment on the antioxidative defense in the skin of rats exposed to heat shock. AB - Skin protection against heat shock and the specificity in the organization of antioxidative defenses were examined in rats given oral antioxidative pretreatment with selenium (Se)-enriched yeast and vitamins E, C, and A for 15 days and then exposed to hyperthermia. The activity of antioxidative enzymes in the skin and the liver was monitored 1 hour and 3 hours after heat shock. Glutathione peroxidase (GSH-Px) activity was increased in the skin after heat shock in the groups supplemented with antioxidants, but not in the controls. In contrast, the activity of liver GSH-Px was increased only in the controls receiving antioxidants. Heat shock led to a decrease in liver superoxide dismutase (SOD) activity at 1 hour in the antioxidant-supplemented group, but this was unchanged in the liver of all other groups and in the skin. The activity of thioredoxin reductase (TR) in the skin was increased in the antioxidant supplemented group 1 hour after heat shock, whereas the hepatic thioredoxin reductase activity was decreased. The activities of catalase (CAT), glutathione reductase (GR), and glutathione-S-transferase (GST) were unaffected by either treatment. These results suggest that supplementation with antioxidants protects the skin against heat shock, especially with respect to the GSH-Px and TR activity. The different response of the skin in comparison with the liver probably reflects differences in organization and regulation of antioxidative defenses. PMID- 9726808 TI - Effects of cereal supplementation with selenium. AB - Selenium (Se) is an essential nutrient. Surveys of Se status showed that Serbia is a low Se area. To increase the Se concentration, we supplemented Se to domestic animals and humans. The latest results showed that increasing Se content in domestic animals and humans induces a small increase in mean Se level. Exceptions were residents of urban, economically developed communities in which an increase in Se level were significant. The soil-plant system is the main natural source of Se for animals and humans. We used field treatments to raise the Se level in plants to the desired level. Foliar application of Se as Na2SeO3 was tested on cereals growing on different soil types. The influence of different growth factors on the uptake and distribution of Se in plants was tested and discussed. The foliar application of 6 g Se/ha is sufficient to raise the native Se content in whole wheat to levels of 42 to 67 microg/kg and in corn to levels of 19 to 36 microg/kg. The application of 12 g Se/ha is sufficient to raise Se levels in wheat to 65 to 180 microg/kg and in corn to 31 to 46 microg/kg. PMID- 9726809 TI - Selenium-dependent and selenium-non-dependent glutathione peroxidase in patients with Balkan endemic nephropathy. AB - We studied the activity of erythrocyte selenium (Se)-dependent, Se-non-dependent glutathione peroxidase (GSH-Px), and superoxide dismutase (SOD) in uremic patients (UP) in clinically healthy members from families affected with Balkan nephropathy (HMF/BEN) and in healthy volunteers from endemic settlements (control group). The SOD activity was not significantly different in the groups studied and the Se-non-dependent GSH-Px activity in HMF/BEN and UP was not different from the control group. However, the activity of Se-dependent GSH-Px in UP was lower compared with the control group, whereas the mean value of the Se-dependent GSH Px activity in HMF/BEN was not significantly different when compared with the other two investigated groups. PMID- 9726810 TI - Glutathione peroxidase in amyotrophic lateral sclerosis: the effects of selenium supplementation. AB - The activity of glutathione peroxidase (GSH-Px) as well as the activities of other antioxidative enzymes: CuZn superoxide dismutase (CuZn SOD), catalase (CAT), glutathione reductase (GR) in erythrocytes, as well as the activity of plasma glutathione transferase (GST), and the plasma content of vitamins E and C were evaluated in 35 sporadic amyotrophic lateral sclerosis (sALS) patients. The results revealed significantly decreased activity of both GSH-Px and CuZn SOD in sALS patients compared with the control. These data showed that a disturbed oxidative/antioxidative balance in sALS patients exists not only in motoneurons but also in the blood. The effect of exogenously administered selenium (Se), antioxidants, amino acids, a Ca2+ channel blocker such as nimodipine, and their combination in Alsamin was evaluated by screening parameter levels after 9 weeks of treatment. Only the use of all components together enhanced the activity of GSH-Px and the amount of vitamin E in sALS patients. Judging by the results of clinical trials, this treatment slowed the course of the disease. PMID- 9726811 TI - The influence of selenium and neuroleptics on the rat brain nuclear regulatory mechanism in carcinogenesis. AB - The potential clinical benefit of the antitumor effect of selenium (Se) has recently been confirmed in tumor-bearing animals and human tumor cells in culture. In clinical medicine, the reduced incidence of cancer among schizophrenic patients was attributed to neuroleptic medication. However, there has been little information on the effect of Se, carcinogen, and neuroleptic on the brain cells. This investigation was carried out on the brains of male Wistar rats treated with inorganic Se, 9,10-dimethyl-1,2-benzanthracene, and chlorpromazine. Chromatin was prepared and purified from isolated brain cell nuclei. Various protein species (histones and nonhistone proteins), RNA, and DNA were extracted by different extraction procedures. A higher relative content of nonhistone proteins was found in the group of animals treated with Se alone, carcinogen alone, and Se plus carcinogen administered simultaneously when compared with other experimental and control groups. The ratio of nonhistone proteins and histones of < 1.0 in the group of animals treated with neuroleptic + carcinogen or with neuroleptic indicates a lower content of nonhistone proteins when compared to histones. We obtained a more pronounced susceptibility to degradation by DNase I in the group of animals treated with neuroleptic + carcinogen or with neuroleptic compared to chromatin in the animals treated with carcinogen alone. We conclude that neuroleptic increases protein synthesis, as well as chromatin susceptibility to enzymatic degradation, thus achieving an opposite effect of carcinogen. PMID- 9726812 TI - Reactive oxidants from nitric oxide, oxidants and cellular signalling, and repair of oxidative DNA damage: a Chemical Pathology Study Section workshop. PMID- 9726813 TI - Platelet-activating factor and the growth of fibroblasts. PMID- 9726814 TI - Distal chromosome 17q loss in Barrett's esophageal and gastric cardia adenocarcinomas: implications for tumorigenesis. AB - The molecular genetic mechanisms underlying esophageal cancer are poorly understood. However, a novel gene that may be involved in esophageal carcinogenesis was recently localized by others to distal 17q by linkage analysis of kindreds with palmoplantar keratoderma and squamous cell carcinoma of the esophagus. To help determine whether a distal 17q gene may also be involved in the pathogenesis of primary Barrett's esophageal and gastric cardia adenocarcinomas, we performed loss of heterozygosity (LOH) analysis of 21 Barrett's and 18 gastric cardia adenocarcinomas at loci spanning 17q: cen-BRCA1 SSTR2-D17S2058-D17S929-D17S722-+ ++D17S937-D17S802-tel. Over 50% of the Barrett's and cardia adenocarcinomas demonstrated loss of an allele at one or more informative distal 17q markers. One common overlapping region of loss involved loci mapped to distal 17q24-proximal 17q25, which tentatively defines a potential chromosomal region distal to BRCA1 involved in the pathogenesis or progression of both types of adenocarcinomas. LOH analysis of DNA from matched microdissected sections of Barrett's metaplasia suggested that loss of D17S2058 in this region may be an early event in the malignant transformation of Barrett's metaplasia. No statistically significant correlations between 17q LOH and tumor stage or patient survival were noted. In summary, LOH mapping of 17q in Barrett's and cardia adenocarcinomas suggests the existence of at least one putative distal 17q tumor suppressor gene involved in the pathogenesis of these tumors. PMID- 9726815 TI - Expression of cytochrome P450 2A5 in preneoplastic and neoplastic mouse liver lesions. AB - Cytochrome P450 (CYP) 2A5 is involved in the metabolism of carcinogens like aflatoxin B1 and N-nitrosodiethylamine (NDEA), and CYP2A5 levels are increased in some pathological states of the liver (e.g., infectious hepatitis and porphyria). We analyzed the expression of CYP2A5 during experimental liver carcinogenesis in three different mouse strains (C3H/He, C57BL/6J, and B6C3F1) with immunohistochemical techniques and in situ hybridization. In normal liver, CYP2A5 protein and mRNA were detected in centrilobular hepatocytes only. Phenobarbital treatment increased the number of CYP2A5-positive centrilobular hepatocytes and the CYP2A5-positive areas were extended into the middle zone in all strains, but periportal hepatocytes remained negative. Fifty percent of the spontaneous foci in untreated mice, over 90% of the foci in mice treated with NDEA or phenobarbital and all of the hepatocellular adenomas and carcinomas displayed positive immunostaining and a strong CYP2A5 mRNA signal by in situ hybridization. In the liver tumors metastasized to the lung, expression of CYP2A5 had largely disappeared. CYP2A5 expression in neoplastic and putative preneoplastic lesions, although sometimes heterogeneous, was apparently independent of the typical zonal expression pattern in normal tissue. As expected, the C57BL/6J mice developed fewer foci and tumors than the C3H/He and B6C3F1 mice, but the phenotype of CYP2A5 overexpression was similar in all the strains. Our data suggest that the increased expression of CYP2A5 may play an important role in the development of liver cancer in mice and may be used as a novel marker for spontaneous and NDEA induced mouse liver foci. PMID- 9726817 TI - RRR-alpha-tocopheryl succinate induction of prolonged activation of c-jun amino terminal kinase and c-jun during induction of apoptosis in human MDA-MB-435 breast cancer cells. AB - We have demonstrated that RRR-alpha-tocopheryl succinate (10 microg/mL vitamin E succinate (VES) treatment of estrogen receptor-negative MDA-MB-435 human breast cancer cells induces 9, 19, 51, and 72% apoptotic cells on days 1-4, respectively, after treatment, which involves transforming growth factor-beta signaling. Here, we show that VES-triggered apoptosis of MDA-MB-435 cells induced prolonged elevated expression of c-jun mRNA and protein (neither of which was caused by major increases in stability) and also induced enhanced activator protein-1 (AP-1) binding to the consensus DNA oligomer. Furthermore, VES treatments resulted in increased AP-1 transactivation activity, as measured with an AP-1 promoter/luciferase reporter construct and by the measurement of increased mRNA expression of the AP-1-dependent endogenous gene collagenase. Evidence of VES-induced involvement of the c-jun amino-terminal kinase in these AP-1-dependent events was suggested by data showing prolonged activity of this kinase, as measured by a kinase assay using glutathione S-transferase-c-jun as the substrate. The c-jun-dependent transcriptional activity was verified by cotransfection of a chimeric transcription factor having a galactose 4 DNA binding domain coupled with the transactivation domain of c-jun plus the reporter plasmid 5X GAL4-luciferase. MDA-MB-435 cells infected with an adenovirus expression vector containing the TAM-67 sequence for dominant/negative-acting mutant c-jun or transiently transfected with c-jun antisense exhibited a 50-77% reduction in VES-mediated apoptosis as compared with control adenovirus-infected or control sense oligomer-transfected cells. PMID- 9726816 TI - Induction of apoptosis by thiuramdisulfides, the reactive metabolites of dithiocarbamates, through coordinative modulation of NFkappaB, c-fos/c-jun, and p53 proteins. AB - Prolinedithiocarbamate (PDTC) and diethyldithiocarbamate (DDTC) are cancer chemopreventive agents and can be biotransformed to prolinethiuramdisulfide (PTDS) and tetraethylthiuramdisulfide (disulfiram; DTDS), respectively. We found that the reactive metabolites PTDS and DTDS induced apoptosis after G1/S arrest. Phosphorylation of cyclin E, inhibition of cyclin-dependent kinase 2 activity, and degradation of cyclin E were found in human hepatoma Hep G2 cells during apoptosis. Moreover, PTDS and DTDS decreased the level of bcl-2 but increased the level of p53. In contrast, PDTC, DDTC, and ammonium dithiocarbamate (ADTC) did not induce apoptosis; rather they led to the induction of p53 and p21 followed by G1/S arrest. PDTC, DDTC, and ADTC also arrested cells in G1 phase. We then examined the effects of PTDS and DTDS on the signal transduction mechanisms leading to apoptosis. Although the transcription factors NFkappaB and AP-1 cooperatively decreased their DNA-binding activities to kappaB and 12-O tetradecanoylphorbol-13-acetate-responsive elements, respectively, and p53 increased DNA-binding activity in the early stage but decreased it in the latter stage after treatment with PTDS, when the human Hep G2 cells were undergoing apoptosis. In summary, our results indicated that (i) PTDS and DTDS induced apoptosis and G1/S arrest mediated by p53, whereas PDTC, DDTC, and ADTC induced p53-dependent p21 expression leading to G1/S arrest; (ii) PDTC, DDTC, and ADTC induced p21/KIP1/CIP1 expression in a p53-dependent pathway leading to G1/S arrest; and (iii) NFkappaB, AP-1, and bcl-2 were downregulated during PTDS- and DTDS-induced apoptosis. These results suggested that PTDS and DTDS induced p53 dependent apoptosis, whereas PDTC, DDTC, and ADTC induced G1/S arrest. Apoptosis is regulated by the modulation of intracellular effectors such as NFkappaB, AP-1, and bcl-2 and activation of p53 in early stages. PMID- 9726818 TI - Ki-ras mutation and cell proliferation of lung lesions induced by 1-nitropyrene in A/J mice. AB - In this study, lung lesions were found in male A/J mice 24 wk after intraperitoneal injection of 1-nitropyrene (1-NP). The lesions were classified into three categories: alveolar/bronchiolar hyperplasia, adenoma, and adenocarcinoma. The proliferation kinetics of cells in the lesions were evaluated by assessing proliferating cell nuclear antigen (PCNA) expression and silver staining nucleolar organizer regions (AgNORs). Furthermore, the role of the Ki ras gene in tumorigenesis was studied by detecting point mutations in Ki-ras codons 12, 13, and 61 by polymerase chain reaction and sequence analysis. The PCNA-positive rates (+/- standard deviations) in various samples were as follows: 0% for specimens from six untreated animals and six uninvolved areas, 4.26 +/- 3.94% for 19 hyperplasias (hyperplasias vs normal lung tissue, P < 0.01), 13.24 +/- 6.35% for 25 adenomas (adenomas vs hyperplasias, P < 0.01), and 38.0 +/- 9.63% for four adenocarcinomas (adenocarcinomas vs adenomas, P < 0.01). The corresponding mean AgNOR scores were as follows: 1.10 +/- 0.05 for the untreated animals, 1.32 +/- 0.09 for the uninvolved areas, 1.72 +/- 0.59 for the hyperplasias (hyperplasias vs normal lung tissue, P > 0.05), 2.74 +/- 0.70 for the adenomas (adenomas vs hyperplasias, P < 0.01), and 5.22 +/- 0.62 for the adenocarcinomas (adenocarcinomas vs adenomas, P < 0.01). Ki-ras gene mutations were identified in three of four (75%) adenocarcinomas, six of 23 (26%) adenomas, and two of 17 (12%) hyperplasias. No mutations were found in normal lung tissue. The most frequent Ki-ras mutation was an arginine (CGA)AT --> GC transition at codon 61 in exon 2. The PCNA-positive rates and AgNOR scores of cases with Ki-ras mutations were higher than those without an identified mutation (P < 0.05). Ki ras mutations at codon 61 (Arg) may therefore influence the growth or development of 1-NP-induced lung lesions in A/J mice. PMID- 9726819 TI - Persistence of an encephalitogenic T cell clone in the spinal cord during chronic, relapsing experimental autoimmune encephalomyelitis. AB - The CDR3 region of the TCR beta-chain of a CD4+, Th1, Vbeta2+ encephalitogenic T cell clone was used as an idiotypic marker to track the location of the clone in vivo. cDNA prepared from the spinal cord, thymus, lymph nodes, spleen, and liver of the recipients at various stages of EAE was amplified using Vbeta2 and Cbeta region primers, and the products immobilized. The membrane was probed with a 32P labeled oligonucleotide complementary to the CDR3 region of the T cell clone. The probe reacted strongly with products from the spinal cord, spleen and liver and less strongly with products from lymph nodes and thymus of mice with acute EAE. The signal was greatly diminished in the spinal cord and other tissues during recovery from acute disease and reappeared in the spinal cord at each relapse. PMID- 9726820 TI - Targeting the B7/CD28:CTLA-4 costimulatory system in CNS autoimmune disease. AB - The B7/CD28:CTLA-4 costimulatory pathway plays a critical role in determining the fate of immune responses (activation vs. down-regulation) and is a highly promising therapeutic target for treating autoimmune diseases. In this review, we highlight the mechanisms by which this costimulatory pathway operates emphasizing the role of the different components in the pathogenesis of relapsing experimental autoimmune encephalomyelitis, a CD4 T cell-mediated autoimmune model of multiple sclerosis. The separate and distinct roles of B7-1, B7-2 and CTLA-4 in positive and negative regulation of autoimmune pathogenesis are considered and a working model is proposed. PMID- 9726821 TI - The steroid hormone dehydroepiandrosterone (DHEA) breaks intranasally induced tolerance, when administered at time of systemic immunization. AB - The induction of intranasal tolerance might be dependent on specific characteristics of mucosal, nose-draining lymph nodes. Such a specific characteristic might lie in the metabolism of the steroid hormone DHEA. Conversion of the prohormone DHEAS into DHEA is dependent on DHEAS-sulphatase activity in lymph nodes. This activity is low in mucosa-draining lymph nodes compared to peripheral lymph nodes, leading to differences in microenvironment. However, administration of DHEA before the induction of intranasal tolerance, could not change tolerance induction. We next determined the effect of DHEA after the induction of intranasally induced tolerance and demonstrated that the steroid hormone and some of its derivatives are able to break tolerance, when administered at time of systemic immunization. These findings might have implications for the regulation of intranasal tolerance and the use of DHEA. PMID- 9726822 TI - Localization of cyclooxygenase-2 and prostaglandin E2 receptor EP3 in the rat lumbar spinal cord. AB - Cyclooxygenase-2 (COX-2) is now considered to be the major constitutively expressed COX isozyme in the central nervous system. The present immunocytochemical study details localization of COX-2 immunoreactivity in rat spinal cord along with the expression of prostaglandin E2 receptor subtype EP3. Prominent COX-2 staining was observed in the nuclear envelope of neurons throughout the spinal cord, especially in the superficial dorsal horn laminae and motoneurons of lamina IX, as well as in glial cells of the white matter. Expression of EP3 receptor was strictly confined to afferent terminal areas in the superficial dorsal horns. PMID- 9726823 TI - A new anti-rheumatic drug, T-614, effectively suppresses the development of autoimmune encephalomyelitis. AB - In the present study, we examined the therapeutic effects of T-614 (3-formylamino 7-methylsulfonylaminoxy-4H-1-benzopyran-4-one), a new anti-rheumatic drug, on a T cell-mediated autoimmune disease, experimental autoimmune encephalomyelitis (EAE). T-614 dose-dependently suppressed the development of active EAE induced in Lewis rats by immunization with myelin basic protein (MBP) when administered for 2 weeks starting on the day of immunization (day 0 to 14). Amelioration of clinical signs was also obtained by the treatment at the effector phase (day 7 to 14) of the disease. Furthermore, T-614 treatment of recipient rats that had received MBP-sensitized lymphoid cells resulted in suppression of the clinical severity of EAE. Immunohistological examination revealed that the number of TCR alpha beta-expressing T cells and the extent of MHC class II expression in the spinal cord of rats treated with T-614 was markedly reduced. In vitro study using MBP-specific T cells showed that the addition of T-614 inhibited the proliferative responses of T cells and the production of pro-inflammatory cytokines such as IFN-gamma, IL-6 and TNF produced by T and accessory cells. Taken together, these findings imply that T-614 suppresses the development of EAE by inhibiting the proliferation of autoreactive T cells and pro-inflammatory cytokine production not only by T cells but also by macrophages/microglia. This may be attributable to the result that T-614 is more effective at the effector phase rather than the induction phase. Thus, this drug has a potential value for the treatment of various T cell-mediated autoimmune diseases including multiple sclerosis (MS) as well as rheumatoid arthritis. PMID- 9726824 TI - Increased vascular endothelial growth factor (VEGF) and transforming growth factor beta (TGFbeta) in experimental autoimmune uveoretinitis: upregulation of VEGF without neovascularization. AB - Experimental autoimmune uveoretinitis (EAU) was induced in Lewis rats and B10.A mice by immunization with S-antigen (S-Ag) to study the potential roles of vascular endothelial growth factor (VEGF) and the beta1 and beta2 isoforms of transforming growth factor (TGFbeta1 and TGFbeta2) during the progression of the disease. VEGF has been implicated as an angiogenic factor in ischemic retinopathies; however, Lewis rats developing EAU have high levels of VEGF in the retina, but no neovascularization. In the present study, immunohistochemical staining for VEGF, TGFbeta1 and TGFbeta2 was performed on the retinas of Lewis rats developing EAU or with oxygen-induced ischemic retinopathy. In rats immunized with S-antigen, a marked upregulation of VEGF was immunohistochemically visualized from the inner nuclear layer to the inner limiting membrane prior to blood-retinal barrier (BRB) failure and lymphocytic infiltration. VEGF is normally induced by hypoxia and its induction leads to neovascularization. Coincident with the increase in VEGF, there was increased immunoreactivity for TGFbeta1 and TGFbeta2 within the same layers of the retina. In contrast, rats with ischemic retinopathy and retinal neovascularization showed only a modest increase in VEGF immunoreactivity, which is largely confined to retinal ganglion cells and inner retinal vessels, and little or no increase in TGFbeta1 or TGFbeta2. In addition, in mice developing EAU, which does not have an abrupt onset as it does in rats and may involve neovascularization, a comparable upregulation of VEGF in the inner retina to that seen in rats developing EAU occurs with no increase in TGFbeta1 or TGFbeta2. Since TGFbeta can inhibit endothelial cell proliferation, it is likely that an increase in TGFbeta may prevent VEGF from exerting its endothelial growth activity in the rat EAU model, but VEGF may be operative in inducing BRB failure. These data suggest that there is a complex interaction among growth factors in the retina and that retinal neovascularization may require an imbalance between stimulatory and inhibitory factors. PMID- 9726825 TI - Stress-related modulation of central nervous system immunity in a murine model of herpes simplex encephalitis. AB - This study assesses the immunomodulatory effects of stress on the pathogenesis of herpes simplex encephalitis (HSE) in a mouse model. Physical restraint served as the stressor and HSE developed subsequent to HSV-1 inoculation into the tongues of subject animals. Clinical data showed that stressed mice lost more weight and had greater mortality rates than unrestrained animals during the course of infection. Histologic tissue sections demonstrated a stress-related reduction of the cellular inflammatory response in the central nervous system (CNS). This model may be useful to further investigate the mechanisms of stress-related immunosuppression in the CNS. PMID- 9726827 TI - Analysis of immune lesions in neurocysticercosis patients: central nervous system response to helminth appears Th1-like instead of Th2. AB - Neurocysticercosis (NCC) caused by the helminth Taenia solium is the most common parasitic infection of the human central nervous system (CNS) worldwide. Because clinical symptoms are associated with localized immunological responses in the brain, characterization of these responses are pivotal for understanding the pathogenesis of cysticercosis. Immunohistochemical analysis of brain specimens from several patients with cysticercosis revealed at least four types of immune responses, including: (i) an antibody response (IgM + plasma cells), (ii) a predominant NK response, (iii) an infiltrate with abundant macrophages and granulocytes, and (iv) an intense infiltrate with a predominance of macrophages and T cells. The intensity and type of immunity appeared to be associated somewhat with the parasite's viability and anatomical location. In most of the lesions, cell mediated responses were evident and proinflammatory cytokines including IL12 predominated. Moreover, IL4 was undetectable in the immune infiltrates. Thus, the CNS response to this helminth, unlike the systemic response, is predominately Th1-like. PMID- 9726826 TI - Identification of haemopoietic biglycan in hyperplastic thymus associated with myasthenia gravis. AB - Generally biglycan, a small proteoglycan, has been thought to play a role as an extracellular matrix and/or a reservoir for other factors, such as TGF-beta and collagens. Recently, we have found that a soluble 100 kDa biglycan, produced from the rat thymic myoid cells and the brain glial cells, predominantly stimulates growth and differentiation of monocytic lineage cells from various lymphatic organs, including microglias. In the present study, we attempted to identify biological significance of the corresponding molecules in human, using five myasthenic thymuses (three with hyperplasia and two with thymoma) that had been surgically removed for therapeutic purpose. With immunohistochemistry, many biglycan positive cells were detected in the germinal center of the three hyperplastic thymuses, but not in the two thymuses associated with lymphocytic thymoma. Biglycan purified from the hyperplastic thymuses by an immunoaffinity column was found as a monomer with apparent molecular size of 95-100 kDa and self associated oligomers of greater than 201 kDa. The purified biglycan markedly stimulated the growth and differentiation of monocytic cells from haemopoietic stem cells of the rat bone marrow. These results suggest that particular cells, which produce haemopoietic biglycan, play significant roles in generation and maintenance of the hyperplastic changes associated with myasthenia gravis. PMID- 9726829 TI - Langerhans cells in the neuro-immuno-cutaneous system. AB - Langerhans cells are epidermal antigen presenting cells. They are able to express some neuronal markers. They express many neuropeptide receptors. Neuropeptides are able to modulate Langerhans cell functions. Langerhans cells are closely connected with nerve fibers in the epidermis. Thus, Langerhans cells and the nervous system are anatomically and functionally associated. We suggest that they belong to a neuroimmunocutaneous system. The interactions between nervous and immune system are probably important in the pathophysiogeny of inflammatory dermatoses. PMID- 9726828 TI - Differential recognition of MBP epitopes in BALB/c mice determines the site of inflammatory disease induction. AB - Although myelin basic protein (MBP)-recognizing T cells are not readily obtained after immunization of BALB/c mice with MBP (reflecting the BALB/c resistance to actively induced experimental autoimmune encephalomyelitis (EAE)), they can be expanded and cloned after several rounds of in vitro culture. The majority of BALB/c-derived clones recognize an epitope defined by MBP peptide 59-76. When transferred to naive BALB/c recipients, these clones cause classical EAE, with characteristic inflammation and demyelination of the central nervous system (CNS). We previously showed that two related clones recognizing a minor epitope, defined by MBP peptide 151-168, cause inflammation and demyelination preferentially of the peripheral nervous system (PNS). Because MBP has alternatively spliced isoforms, residues 151-168 are not present contiguously in all MBP isoforms. In order to determine whether induction of PNS disease is idiosyncratic to these sister clones, or related to their properties of epitope recognition, an independent T-cell line with similar recognition properties was studied. Clone 116F, derived from a BALB/c shiverer mouse, expresses a different T-cell receptor (TCR), with distinct TCR contact residues, but like the previously described T cells, this clone requires residues from both exons 6 and 7 for optimal stimulation. When adoptively transferred to BALB/c recipients, this clone preferentially induces disease of the PNS. A control BALB/c shiverer derived MBP 59-76-recognizing clone, in contrast, induces CNS disease. These data strongly suggest that the site of disease initiation may correlate with epitope recognition, particularly when alternative isoforms are involved. PMID- 9726830 TI - Accumulation of passively transferred primed T cells independently of their antigen specificity following central nervous system trauma. AB - The central nervous system (CNS) enjoys a unique relationship with the immune system. Under non-pathological conditions, T cells move through the CNS but do not accumulate there. CNS trauma has been shown to trigger a response to CNS self antigens such as myelin basic protein (MBP). Here, we examined whether the injured CNS tissue undergoes changes that permit T cell accumulation. We found that injury to CNS white matter, such as the optic nerve, led to a transiently increased accumulation of T cells (between days 3 and 21). In Lewis rats with unilaterally injured optic nerves, systemic administration of passively transferred T cells recognizing either self-antigen (MBP) or non-self-antigen (ovalbumin) resulted in accumulation of the T cells in injured optic nerve, irrespective of their antigenic specificity. The effect of the T cells on the damaged nerve, the lack of selectivity in T cell accumulation and the mechanism underlying non-selective accumulation are discussed. PMID- 9726831 TI - Concentration-dependent effects of pentoxifylline on migration and myelin phagocytosis by macrophages. AB - The effects of pentoxifylline (POX) on macrophage migration and myelin uptake were studied in an in vitro model of myelin phagocytosis. The POX is a phosphodiesterase inhibitor which inhibits TNF-alpha (tumor necrosis factor alpha) production and reduces ICAM-1 (intercellular adhesion molecule-1) expression by macrophages. Both of these molecules have earlier been shown to be involved in the process of myelin recognition and degradation. In the present series of experiments, cocultured peripheral nerves and macrophages were treated with different concentrations of POX. Untreated controls were massively invaded by macrophages which ingested the degenerating myelin sheaths. High concentrations of POX (100 microg ml(-1)) inhibited macrophage invasion of the nerves. Lower POX concentrations (50 microg ml(-1)), in contrast, lead to an increased myelin uptake by phagocytic cells without affecting macrophage migration. These data indicate that POX may regulate different effector functions of macrophages such as migration and myelin phagocytosis during Wallerian degeneration. This is important for inflammatory demyelinating conditions in the central or peripheral nervous system (PNS) in which macrophages are also important effector cells. Since POX is used as an immunomodulatory drug in demyelinating diseases, its effects on the described macrophage functions may be of high relevance. An increased myelin uptake during Wallerian degeneration may also support a more efficient axonal regeneration by removing axonal outgrowth inhibitors. PMID- 9726832 TI - Influence of adhesion molecule expression by human brain microvessel endothelium on cancer cell adhesion. AB - Cultures of endothelial (En) cells derived from human brain microvessels were established in order to characterize adhesion molecule expression and to assay the adhesion properties of neoplastic cell lines to monolayers of En cells. Low constitutive expression of beta1 integrin (CD29), and ICAM-2 (CD102) was detected on human brain microvessel En cells. The beta1 chain of the VLA integrin family, ICAM-1, E-selectin (CD62E) and VCAM-1 (CD106) but not ICAM-2 and PECAM-1 (CD31) expression was upregulated by IL1-alpha, and TNF-alpha proinflammatory cytokines. High expression of PECAM-1 was found on non-activated human brain EN cells. In order to study the potential role of adhesion molecules in neoplastic cell adhesion two tumor cell lines were chosen. Adhesion of a cell line (DU145) derived from a cerebral metastasis of prostate carcinoma to human brain microvessel En cell monolayers was less pronounced compared to adhesion of a primary prostate carcinoma cell line (ND1). Adhesion of cerebral metastatic neoplastic cell line (DU145) was not significantly influenced by incubation of endothelial cells with different proinflammatory cytokines. The adhesion capability of primary prostate carcinoma line (NDI) was significantly upregulated by TNF-alpha proinflammatory cytokine. Furthermore, the adhesion of ND1 was partly inhibited using anti-E-selectin and VCAM-1 monoclonal antibodies. There was no significant effect of anti-adhesion antibodies on the adhesion characteristics of the cerebral metastatic (DU145) cell line. Our data demonstrate that different mechanisms are involved in the adhesion of neoplastic cells to cerebral En cells and turn our attention to the importance of adhesion molecule expression in the formation of metastases. PMID- 9726833 TI - Astrocytes induce hyporesponses of myelin basic protein-reactive T and B cell function. AB - We have isolated infiltrating mononuclear cells (inMNC) and lymph node MNC (lnMNC) from Lewis rats with actively induced experimental allergic encephalomyelitis (EAE), and compared their responses to myelin basic protein (MBP). MBP-induced proliferation and MBP-specific IgG secreting cells were lower in inMNC compared to lnMNC, while MBP-reactive IFN-gamma secreting cells in inMNC were higher. Using an in vitro culture system, we observed that astrocytes derived from newborn Lewis rats not only suppressed T cell proliferation and IFN gamma production but also reduced MBP-specific IgG production by B cells. Astrocyte-derived soluble factor (s), rather than direct cell-to-cell interactions, seems to be responsible for the inhibitory effect. Supernatants from IFN-gamma-stimulated astrocytes exhibited stronger suppressive effects than supernatants from unstimulated astrocytes, whereas supernatants from microglia did not cause downregulation of T and B cell functions. These results indicate that astrocytes are major effector cells in inhibiting functions of MBP-reactive T and B cells. Astrocytes down-regulated expression of ICAM-1 and MHC class II in lnMNC, but did not induce apoptosis of lnMNC. The results in the present study do not exclude the possibility that astrocytes induce T cell apoptosis in a cognate fashion. Taken together, the inhibitory properties of astrocytes may contribute to the confinement of inflammatory lesions in multiple sclerosis and EAE. Our report compares immunoreactivities of inMNC and lnMNC in EAE and elucidates the role of astrocytes in the inactivation of MBP-reactive T and B cells. PMID- 9726834 TI - Reciprocal stimulation between TNF-alpha and nitric oxide may exacerbate CNS inflammation in experimental autoimmune encephalomyelitis. AB - Nitric oxide (NO) and TNF-alpha are both highly active pleotypic modulators of cell function that are abundantly generated during inflammation. Experiments in animal systems have linked the generation of NO and TNF-alpha to autoimmune pathogenesis, and blockade of either NO or TNF-alpha has been shown to impede disease development. In this study, we show that NO and TNF-alpha can act mutually to stimulate each other's production. While IFN-gamma alone induces NO release from microglia, astrocytes are provoked into significant NO production only by a combination of IFN-gamma and TNF-alpha. Since both TNF-alpha and NO are abundantly generated during T-glial cell interaction, we asked whether and how NO affects TNF-alpha production. Using an in vitro system in which TNF-alpha secretion is induced in MBP-reactive T cells by co-culture with syngeneic astrocytes, we observed that the efficiency of TNF-alpha secretion was markedly increased, in a dose-dependent fashion, by addition of micromolar concentrations of a chemical generator of NO donor, sodium nitroprusside (SNP). Similarly, low concentrations of SNP significantly enhanced the IL-2 dependent growth of MBP reactive T cells. These results suggest that autoimmune pathogenesis initiated by inflammatory responses within the CNS may result in part from a vicious cycle in which TNF-alpha and NO mutually provoke each other's production. PMID- 9726835 TI - Role of the target organ in determining susceptibility to experimental autoimmune myasthenia gravis. AB - Injection of anti-AChR antibodies in passive transfer experimental autoimmune myasthenia gravis (EAMG) results in increased degradation of acetylcholine receptor (AChR) and increased synthesis of AChR alpha-subunit mRNA. Passive transfer of anti-Main Immunogenic Region (MIR) mAb 35 in aged rats does not induce clinical signs of disease nor AChR loss. The expression of the AChR subunit genes was analyzed in susceptible and resistant rats. In aged EAMG resistant rats, no increase in the amount of AChR alpha-subunit mRNA was measured. In vivo AChR degradation experiments did not show any increase in AChR degradation rates in aged resistant rats, in contrast to young susceptible rats. Taken together, these data demonstrate that resistance of the AChR protein to antibody-mediated degradation is the primary mechanism that accounts for the resistance to passive transfer EAMG in aged rats. PMID- 9726836 TI - Induction of cyclooxygenase (COX)-2 but not COX-1 gene expression in apoptotic cell death. AB - In this study we assessed the regulation of cyclooxygenase (COX)-2 in models of apoptotic cell death in vivo and in vitro. By 6 h after hippocampal colchicine injection in rat, COX-2 (but not COX-1) mRNA expression was elevated. The induction of COX-2 mRNA expression preceded temporally and overlapped anatomically the cellular morphological features of apoptosis in the granule cell layer of the dentate gyrus. Similarly, in an established in vitro model of apoptosis in P19 cells, COX-2 induction preceded apoptosis in response to serum deprivation by 12 h. These studies suggest that COX-2 may be involved in the early mechanisms leading to apoptosis. PMID- 9726838 TI - Increased levels of IFN-gamma in the trigeminal ganglion correlate with protection against HSV-1-induced encephalitis following subcutaneous administration with androstenediol. AB - Androstenediol (AED) is a metabolic product of dehydroepiandrosterone (DHEA), an adrenal steroid known to possess immunomodulatory characteristics. The present study was undertaken to assess the efficacy of AED treatment in mice ocularly infected with herpes simplex virus type 1 (HSV-1). The subcutaneous administration of 320 mg/kg AED 4 h prior to viral inoculation was found to enhance the survival of HSV-1-infected mice while lower doses (32.0-100.0 mg/kg) were without effect. However, there were no apparent differences in the viral load in the eye or trigeminal ganglion (TG) 3 or 6 days post infection (p.i.) in vehicle- or AED (320 mg/kg)-treated mice. Likewise, there were no differences in the expression of cytokine or chemokine mRNAs in the eyes or TG early (i.e., 3 days p.i.) following infection. However, by 6 days p.i., there was a significant increase in the expression of the chemokines IP-10, MCP-1, and RANTES and the cytokines interleukin-6 (IL-6) and interferon-gamma (IFN-gamma) in the AED (320 mg/kg)-treated mice compared to vehicle-treated controls as determined by reverse transcription (RT)-polymerase chain reaction (PCR) and quantitative PCR (for IFN gamma). Likewise, there was a corresponding increase in IFN-gamma and IL-2 but not IL-12 protein in the TG of AED-treated mice 6 days p.i. AED-treatment also induced a rise in splenic natural killer activity in a dose- and time-dependent fashion. Collectively, these results suggest that the protective effect following subcutaneous administration of AED is associated in a rise in selective type 1 cytokines (IL-2 and IFN-gamma) as well as natural killer activity. PMID- 9726837 TI - Role of central mu-opioid receptors in the modulation of nitric oxide production by splenocytes. AB - Previous studies have shown that administration of morphine results in alterations of splenic macrophage nitric oxide production. The present studies were conducted to determine the subtype of opioid receptor involved in the modulation of macrophage nitric oxide production. Moreover, the present work was directed at determining whether nitric oxide production is regulated through opioid receptors in the central nervous system (CNS) or via opioid receptors found directly on splenocytes. The study shows that intracerebroventricular (i.c.v.) administration of the mu-selective opioid agonist, DAMGO, to rats dose dependently increases the production of nitric oxide by splenocytes stimulated with toxic shock syndrome toxin (TSST-1). The effect of DAMGO is blocked by prior i.c.v. administration of N-methylnaltrexone. In contrast, i.c.v. administration of the kappa-selective agonist, U69,593, and the delta-selective agonist, DPDPE, have no significant effect on the production of nitric oxide. Furthermore, the in vitro administration of DAMGO, DPDPE, or U69,593 to splenocytes cultures does not significantly alter the production of nitric oxide by splenocytes. In addition, the present work shows that elevation of nitric oxide production by i.c.v. administration of DAMGO produces functional changes in splenic lymphocytes. Collectively, these results indicate that mu-opioid receptors within the CNS are involved in the regulation of splenic nitric oxide production. PMID- 9726839 TI - Expression of interleukin 6 in the rat striatum following stereotaxic injection of quinolinic acid. AB - Stereotaxic intrastriatal injection of the naturally occurring N-methyl-D aspartate (NMDA) agonist quinolinic acid (QA) serves as a valuable in vivo model to study excitotoxic cell damage in the central nervous system (CNS). Although morphological changes such as neuronal loss, glial activation and remote reactions following QA injection have been described in some detail, much less is known about the molecular mechanisms mediating the accompanying glial response. Cytokines are known to play a crucial role in almost all kinds of CNS alterations. We now demonstrate that IL-6, a multifunctional glycoprotein which belongs to the family of neurokines, is expressed endogenously in the rat striatum following QA injection. Using Northern blot analysis, a massive but transient upregulation of IL-6 mRNA could be detected. This started 3 h after QA injection, reached a maximum at 6 h and disappeared within 24 h. That activated microglia are the most likely cellular source of the observed corresponding IL-6 protein expression could be concluded by comparing the immunocytochemical pattern of IL-6 expression and microglial activation. Interestingly, astrocytes initially downregulate their expression of glial fibrillary acidic protein (GFAP) in the excitotoxically injured striatum, but show a delayed increase in GFAP immunoreactivity starting in the periphery of the striatum, subsequently expanding to the core. The early transient IL-6 expression may play an important role in initiating the delayed astrocytic response following excitotoxic cell injury. PMID- 9726840 TI - Augmented intrathecal secretion of interferon-gamma in response to Borrelia garinii in neuroborreliosis. AB - The Lyme disease agent Borrelia garinii has been suggested to be neurotrop, preferentially affecting the nervous system. We compared the secretion of interferon-gamma in response to outer surface proteins from Borrelia garinii and Borrelia afzelii in 10 patients with neuroborreliosis. In cerebrospinal fluid, stimulation with Borrelia garinii revealed higher numbers of interferon-gamma secreting cells in all patients, whereas in blood, only five displayed higher numbers. This further strengthens the hypothesis of Borrelia garinii being associated with the development of neuroborreliosis. PMID- 9726841 TI - Chemotactic activity and IL-8 levels in the cerebrospinal fluid in canine steroid responsive meningitis-arteriitis. AB - Steroid responsive meningitis-arteriitis (SRMA) is a systemic immune disorder, characterized by inflammatory-stenosing lesions of the meningeal arteries and meningitis. The predilection of the disease for the central nervous system (CNS) remains unexplained. In this study, chemotactic activity and chemotactic factors were measured in the cerebrospinal fluid (CSF) of dogs with SRMA. CSF of dogs with SRMA exerted a marked chemotactic activity for leukocytes. Neutrophils were attracted to a similar degree as by CSF from animals with bacterial encephalitis. Chemotactic activity was also noted for mononuclear cells, however, by far weaker than in CSF from animals with viral encephalitis. While the inflammatory process could be suppressed with glucocorticoid treatment, the chemotactic activity of CSF persisted. We could identify IL-8-like activities using a desensitization assay in the CSF of animals with SRMA and also found increased IgA levels. Increased chemotactic activity for polymorphonuclear leukocytes correlated positively with the levels of IL-8-like activity in CSF. Our observations clearly suggest that in SRMA chemotactic factors are generated in the CNS. These include IL-8, but probably also others. The intensity of this production appears to correlate with IgA levels in the CSF suggesting either a causal link or reflecting the severity of the inflammation. PMID- 9726842 TI - Inhibition of group B streptococcal growth by IFN gamma-activated human glioblastoma cells. AB - Group B streptococci are the most important bacteria inducing neonatal septicemia and meningitis. The aim of this study was to assess the role of IFNgamma in the induction of anti-microbial effector mechanisms in human brain tumor cells. Different human glioblastoma/astrocytoma cell lines, stimulated with IFNgamma, restricted the growth of group B streptococci. In addition, we found that TNF alpha is able to enhance the IFNgamma-mediated anti-microbial effect. In contrast to group B streptococci, other bacteria which are also capable of inducing meningitis, like E. coli and all but one of the tested Streptococcus pneumoniae strains, were not influenced by the IFNgamma treated cells. We found that the IFNgamma or the IFNgamma/TNF alpha induced activation of indoleamine 2,3 dioxygenase is responsible for the inhibition of streptococcal growth, since the addition of supplemental L-tryptophan completely blocks the IFNgamma induced bacteriostasis. PMID- 9726843 TI - Cytokine gene expression in cells derived from CSF of multiple sclerosis patients. AB - Cytokines are produced by numerous cell types in the peripheral blood and central nervous system (CNS), and have been implicated in the immunopathogenesis of multiple sclerosis (MS). We examined the relationship between cytokine gene expression of cerebrospinal fluid (CSF) derived cells from MS patients and disease activity as measured by contrast enhanced MRI. There was a significant correlation between the number of CSF cells and the number of contrast enhancing MRI lesions. Cytokine gene expression of TNF-alpha, IFN-gamma, and IL-10 was routinely seen, but IL-4 expression was absent except in two clinically quiescent patients. Trends were observed toward decreased TNF-alpha expression, but increased IL-10 expression, after treatment with IFN-beta1b. PMID- 9726844 TI - Vasoactive intestinal peptide inhibits IL-12 and nitric oxide production in murine macrophages. AB - Vasoactive intestinal peptide (VIP) is a naturally occurring neuropeptide widely distributed in the nervous system. In this study, we investigated the effect of VIP on IL-12, TNF alpha and nitric oxide (NO) production in macrophages following activation with lipopolysaccharide (LPS) or superantigens. In vitro studies show that at physiologic concentrations, VIP inhibited IL-12 and NO but not TNF alpha production in macrophages which were stimulated with LPS or superantigens. The inhibitory effect of VIP on IL-12 production appeared to be cAMP mediated since other cAMP inducing agents were also potent in inhibiting IL-12 production. Since IL-12 plays a critical role in T cell function, we suggest that naturally occurring neural hormones can regulate the type and direction of the immune response. PMID- 9726845 TI - Addicted women: when your patient can't stop drinking, smoking, shopping, eating. AB - While they do not necessarily present the same health risk, such other problems as compulsive shopping and eating have similarly serious implications for both physical and emotional health. All of these behaviors share a common theme reflecting difficulty with impulse control. In some cases, identifying patients with impulse control problems is straightforward; the pack of cigarettes or nicotine stains are easily visible, or the patient's weight can be measured in the office. Other problems, such as alcohol abuse or compulsive shopping, are less observable. To further compound the difficulty with identification is the fact that physicians may not ask routinely about the existence of such problems. Whether a practitioner inquires about the existence of these or other impulse control problems may be related to the difficulty one faces when the problem is finally identified. Addictive behaviors and impulse control disorders are notoriously difficult to treat. High relapse rates and the frustration of both patient and health care provider when efforts to change fail are a testimonial to the tenacity of these behaviors. Recent research highlighting how to meet patients at their level of readiness to change a risk-related behavior offers a perspective on what clinicians can do to assist their patients in approaching and ultimately changing these behaviors. The stages of change model will be described as a method for conceptualizing how behavior change can be facilitated in patients with these various problems. In addition, issues relevant to making a referral will be discussed. PMID- 9726846 TI - Alternatives to hysterectomy for benign conditions. AB - Hysterectomy is the second most commonly performed major operation in the United States. Approximately one in three women will have this operation, resulting in 590,000 procedures per year. The most common indications for hysterectomy are leiomyomata uteri, abnormal uterine bleeding, endometriosis, pelvic pain, and pelvic organ prolapse. Although hysterectomy is an appropriate therapeutic option for some women with these conditions, in many instances less radical alternatives may be offered. Leiomyomata may be managed expectantly if symptoms are not bothersome; for women with troubling leiomyomata symptoms, alternatives to hysterectomy include: endoscopic removal or destruction of myomas, arterial embolization, or hormonal therapy to inhibit or modify bleeding. Endometriosis and abnormal uterine bleeding of leiomyomata are both amenable to hormonal therapy. Pelvic pain is most effectively approached with a thorough evaluation (particularly for nongynecologic illness), with specific therapy directed at the cause of the pain. Pelvic organ prolapse may respond symptomatically to pelvic floor exercises, or to the use of a pessary. After alternatives to removal of the uterus are discussed, the informed woman may decide that hysterectomy is the option best suited to her. It is unusual for hysterectomy to be her only option. PMID- 9726847 TI - Relationship between creatine kinase activity and semen characteristics in subfertile men. AB - OBJECTIVE: Creatine kinase is an indicator of sperm maturity. We studied whether sperm creatine kinase levels differ between normal healthy donors and subfertile patients and determined the correlation between sperm creatine kinase level and semen quality in subfertile men. MATERIAL AND METHODS: Semen samples were obtained from 76 subfertile and 15 healthy normal donors after 48 to 72 hours of sexual abstinence. Sperm characteristics were assessed with a computer-assisted semen analyzer. Morphology was evaluated by Kruger's strict criteria and World Health Organization methods. The thiobarbituric acid assay was used to measure lipid peroxidation; sperm creatine kinase activity was measured using a commercial kit after detergent extraction (Triton X-100). RESULTS: Creatine kinase levels were significantly higher (P < .001) in subfertile men (median = 0.197 U/10(8) sperm) compared with donors (median = 0.061 U/10(8) sperm). In subfertile men, creatine kinase levels correlated significantly with lipid peroxidation levels (r = .49; P = 0.03) and sperm concentration (r = -.70; P < .001), and with normal sperm forms by Kruger's (r = -.30; P = 0.01) and WHO methods (r = -.32; P < .005). Creatine kinase levels and spermatozoal characteristics did not correlate significantly in donors. Compared with subfertile normospermic men, creatine kinase activity was significantly higher in oligospermic and asthenospermic men (P <.001). CONCLUSIONS: The inverse relationship between creatine kinase level and sperm concentration and morphological forms suggests that creatine kinase levels can be a reliable marker for semen quality in subfertile men. An elevated creatine kinase level and its correlation with lipid peroxidation levels may reflect biochemically immature spermatozoa. PMID- 9726848 TI - An International Symposium on Ovarian Aging and Failure. September 4-5, 1998, Brussels, Belgium. Abstracts. PMID- 9726849 TI - Invasion of protozoa by Legionella pneumophila and its role in bacterial ecology and pathogenesis. PMID- 9726850 TI - Signal transduction in the protozoan host Hartmannella vermiformis upon attachment and invasion by Legionella micdadei. AB - The intracellular pathogens Legionella micdadei and Legionella pneumophila are the two most common Legionella species that cause Legionnaires' disease. Intracellular replication within pulmonary cells is the hallmark of Legionnaires' disease. In the environment, legionellae are parasites of protozoans, and intracellular bacterial replication within protozoans plays a major role in the transmission of Legionnaires' disease. In this study, we characterized the initial host signal transduction mechanisms involved during attachment to and invasion of the protozoan host Hartmannella vermiformis by L. micdadei. Bacterial attachment prior to invasion of H. vermiformis by L. micdadei is associated with tyrosine dephosphorylation of multiple host cell proteins, including a 170-kDa protein. We have previously shown that this 170-kDa protein is the galactose N acetylgalactosamine (Gal/GalNAc)-inhibitable lectin receptor that mediates attachment to and invasion of H. vermiformis by L. pneumophila. Subsequent bacterial entry targets L. micdadei into a phagosome that is not surrounded by the rough endoplasmic reticulum (RER). In contrast, uptake of L. pneumophila mediated by attachment to the Gal/GalNAc lectin is followed by targeting of the bacterium into an RER-surrounded phagosome. These results indicate that despite similarities in the L. micdadei and L. pneumophila attachment-mediated signal transduction mechanisms in H. vermiformis, the two bacterial species are targeted into morphologically distinct phagosomes in their natural protozoan host. PMID- 9726852 TI - Cloning and expression of the Listeria monocytogenes scott A ptsH and ptsI genes, coding for HPr and enzyme I, respectively, of the phosphotransferase system. AB - The phosphoenolpyruvate (PEP)-dependent phosphotransferase system (PTS) utilizes high-energy phosphate present in PEP to drive the uptake of several different carbohydrates in bacteria. In order to examine the role of the PTS in the physiology of Listeria monocytogenes, we identified the ptsH and ptsI genes encoding the HPr and enzyme I proteins, respectively, of the PTS. Nucleotide sequence analysis indicated that the predicted proteins are nearly 70% similar to HPr and enzyme I proteins from other organisms. Purified L. monocytogenes HPr overexpressed in Escherichia coli was also capable of complementing an HPr defect in heterologous extracts of Staphylococcus aureus ptsH mutants. Additional studies of the transcriptional organization and control indicated that the ptsH and ptsI genes are organized into a transcription unit that is under the control of a consensus-like promoter and that expression of these genes is mediated by glucose availability and pH or by by-products of glucose metabolism. PMID- 9726851 TI - Isolation, characterization, and heterologous expression of the novel lantibiotic epicidin 280 and analysis of its biosynthetic gene cluster. AB - Epicidin 280 is a novel type A lantibiotic produced by Staphylococcus epidermidis BN 280. During C18 reverse-phase high-performance liquid chromatography two epicidin 280 peaks were obtained; the two compounds had molecular masses of 3,133 +/- 1.5 and 3,136 +/- 1.5 Da, comparable antibiotic activities, and identical amino acid compositions. Amino acid sequence analysis revealed that epicidin 280 exhibits 75% similarity to Pep5. The strains that produce epicidin 280 and Pep5 exhibit cross-immunity, indicating that the immunity peptides cross-function in antagonization of both lantibiotics. The complete epicidin 280 gene cluster was cloned and was found to comprise at least five open reading frames (eciI, eciA, eciP, eciB, and eciC, in that order). The proteins encoded by these open reading frames exhibit significant sequence similarity to the biosynthetic proteins of the Pep5 operon of Staphylococcus epidermidis 5. A gene for an ABC transporter, which is present in the Pep5 gene cluster but is necessary only for high yields (G. Bierbaum, M. Reis, C. Szekat, and H.-G. Sahl, Appl. Environ. Microbiol. 60:4332-4338, 1994), was not detected. Instead, upstream of the immunity gene eciI we found an open reading frame, eciO, which could code for a novel lantibiotic modification enzyme involved in reduction of an N-terminally located oxopropionyl residue. Epicidin 280 produced by the heterologous host Staphylococcus carnosus TM 300 after introduction of eciIAPBC (i.e., no eciO was present) behaved homogeneously during reverse-phase chromatography. PMID- 9726853 TI - Isolation of Mycobacterium paratuberculosis from milk by immunomagnetic separation. AB - An immunomagnetic separation (IMS) technique was developed to facilitate selective isolation of Mycobacterium paratuberculosis cells from milk. Rabbit polyclonal antibodies against radiation-killed intact M. paratuberculosis cells were produced and used to coat sheep anti-rabbit immunoglobulin G (IgG) type M 280 Dynabeads. The rabbit anti-M. paratuberculosis IgG-coated beads (IMB) reacted strongly with laboratory strains of M. paratuberculosis as determined by slide agglutination, and microscopic examination confirmed that M. paratuberculosis cells attached to the IMB. The IMB were found to have a maximum binding capacity of 10(4) to 10(5) CFU of M. paratuberculosis. Studies showed that IMS selectively recovered M. paratuberculosis from inoculated milk containing as few as 10 CFU of M. paratuberculosis per ml when 10 microliter IMB (ca. 10(6) beads) was added to 1 ml of milk and the preparation was incubated for 30 min at room temperature with gentle agitation. Larger volumes of milk (10 and 50 ml) were centrifuged and resuspended in 1 ml of phosphate-buffered saline-0.05% Tween 20 prior to IMS in order to increase the sensitivity of the method. Currently, primary isolation of M. paratuberculosis from a milk sample relies on chemical decontamination, followed by culturing on Herrold's egg yolk medium, which must be incubated at 37 degreesC for up to 18 weeks. The potential value of our IMS method is as an aid for rapid detection of M. paratuberculosis in milk when it is used in conjunction with end point detection methods, such as IS900 PCR or an enzyme-linked immunosorbent assay. PMID- 9726854 TI - Modeling of the competitive growth of Listeria monocytogenes and Lactococcus lactis in vegetable broth. AB - Current mathematical models used by food microbiologists do not address the issue of competitive growth in mixed cultures of bacteria. We developed a mathematical model which consists of a system of nonlinear differential equations describing the growth of competing bacterial cell cultures. In this model, bacterial cell growth is limited by the accumulation of protonated lactic acid and decreasing pH. In our experimental system, pure and mixed cultures of Lactococcus lactis and Listeria monocytogenes were grown in a vegetable broth medium. Predictions of the model indicate that pH is the primary factor that limits the growth of L. monocytogenes in competition with a strain of L. lactis which does not produce the bacteriocin nisin. The model also predicts the values of parameters that affect the growth and death of the competing populations. Further development of this model will incorporate the effects of additional inhibitors, such as bacteriocins, and may aid in the selection of lactic acid bacterium cultures for use in competitive inhibition of pathogens in minimally processed foods. PMID- 9726856 TI - Purification and characterization of laccase from Chaetomium thermophilium and its role in humification. AB - Chaetomium thermophilium was isolated from composting municipal solid waste during the thermophilic stage of the process. C. thermophilium, a cellulolytic fungus, exhibited laccase activity when it was grown at 45 degreesC both in solid media and in liquid media. Laccase activity reached a peak after 24 h in liquid shake culture. Laccase was purified by ultrafiltration, anion-exchange chromatography, and affinity chromatography. The purified enzyme was identified as a glycoprotein with a molecular mass of 77 kDa and an isoelectric point of 5.1. The laccase was stable for 1 h at 70 degreesC and had half-lives of 24 and 12 h at 40 and 50 degreesC, respectively. The enzyme was stable at pH 5 to 10, and the optimum pH for enzyme activity was 6. The purified laccase efficiently catalyzed a wide range of phenolic substrates but not tyrosine. The highest levels of affinity were the levels of affinity to syringaldazine and hydroxyquinone. The UV-visible light spectrum of the purified laccase had a peak at 604 nm (i.e., Cu type I), and the activity was strongly inhibited by Cu chelating agents. When the hydrophobic acid fraction (the humic fraction of the water-soluble organic matter obtained from municipal solid waste compost) was added to a reaction assay mixture containing laccase and guaiacol, polymerization took place and a soluble polymer was formed. C. thermophilium laccase, which is produced during the thermophilic stage of composting, can remain active for a long period of time at high temperatures and alkaline pH values, and we suggest that this enzyme is involved in the humification process during composting. PMID- 9726855 TI - Analysis of Escherichia coli O157:H7 survival in ovine or bovine manure and manure slurry. AB - Farm animal manure or manure slurry may disseminate, transmit, or propagate Escherichia coli O157:H7. In this study, the survival and growth of E. coli O157:H7 in ovine or bovine feces under various experimental and environmental conditions were determined. A manure pile collected from experimentally inoculated sheep was incubated outside under fluctuating environmental conditions. E. coli O157:H7 survived in the manure for 21 months, and the concentrations of bacteria recovered ranged from <10(2) to 10(6) CFU/g at different times over the course of the experiment. The DNA fingerprints of E. coli O157:H7 isolated at month 1 and month 12 were identical or very similar. A second E. coli O157:H7-positive ovine manure pile, which was periodically aerated by mixing, remained culture positive for 4 months. An E. coli O157:H7-positive bovine manure pile was culture positive for 47 days. In the laboratory, E. coli O157:H7 was inoculated into feces, untreated slurry, or treated slurry and incubated at -20, 4, 23, 37, 45, and 70 degreesC. E. coli O157:H7 survived best in manure incubated without aeration at temperatures below 23 degreesC, but it usually survived for shorter periods of time than it survived in manure held in the environment. The bacterium survived at least 100 days in bovine manure frozen at -20 degreesC or in ovine manure incubated at 4 or 10 degreesC for 100 days, but under all other conditions the length of time that it survived ranged from 24 h to 40 days. In addition, we found that the Shiga toxin type 1 and 2 genes in E. coli O157:H7 had little or no influence on bacterial survival in manure or manure slurry. The long-term survival of E. coli O157:H7 in manure emphasizes the need for appropriate farm waste management to curtail environmental spread of this bacterium. This study also highlights the difficulties in extrapolating laboratory data to on-farm conditions. PMID- 9726858 TI - Localization and solubilization of the Iron(III) reductase of geobacter sulfurreducens AB - The iron(III) reductase activity of Geobacter sulfurreducens was determined with the electron donor NADH and the artificial electron donor horse heart cytochrome c. The highest reduction rates were obtained with Fe(III) complexed by nitrilotriacetic acid as an electron acceptor. Fractionation experiments indicated that no iron(III) reductase activity was present in the cytoplasm, that approximately one-third was found in the periplasmic fraction, and that two thirds were associated with the membrane fraction. Sucrose gradient separation of the outer and cytoplasmic membranes showed that about 80% of the iron(III) reductase was present in the outer membrane. The iron(III) reductase could be solubilized from the membrane fraction with 0.5 M KCl showing that the iron(III) reductase was weakly bound to the membranes. In addition, solubilization of the iron(III) reductase from whole cells with 0.5 M KCl, without disruption of cells, indicated that the iron(III) reductase is a peripheral protein on the outside of the outer membrane. Redox difference spectra of KCl extracts showed the presence of c-type cytochromes which could be oxidized by ferrihydrite. Only one activity band was observed in native polyacrylamide gels stained for the iron(III) reductase activity. Excision of the active band from a preparative gel followed by extraction of the proteins and sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed the presence of high-molecular-mass, cytochrome containing proteins in this iron(III) reductase activity band. From these experimental data it can be hypothesized that the iron(III) reductase of G. sulfurreducens is a peripheral outer membrane protein that might contain a c-type cytochrome. PMID- 9726857 TI - Cloning, nucleotide sequence, and expression in Escherichia coli of levansucrase genes from the plant pathogens Pseudomonas syringae pv. glycinea and P. syringae pv. phaseolicola. AB - Plant-pathogenic bacteria produce various extracellular polysaccharides (EPSs) which may function as virulence factors in diseases caused by these bacteria. The EPS levan is synthesized by the extracellular enzyme levansucrase in Pseudomonas syringae, Erwinia amylovora, and other bacterial species. The lsc genes encoding levansucrase from P. syringae pv. glycinea PG4180 and P. syringae pv. phaseolicola NCPPB 1321 were cloned, and their nucleotide sequences were determined. Heterologous expression of the lsc gene in Escherichia coli was found in four and two genomic library clones of strains PG4180 and NCPPB 1321, respectively. A 3. 0-kb PstI fragment common to all six clones conferred levan synthesis on E. coli when further subcloned. Nucleotide sequence analysis revealed a 1,248-bp open reading frame (ORF) derived from PG4180 and a 1,296-bp ORF derived from NCPPB 1321, which were both designated lsc. Both ORFs showed high homology to the E. amylovora and Zymomonas mobilis lsc genes at the nucleic acid and deduced amino acid sequence levels. Levansucrase was not secreted into the supernatant but was located in the periplasmic fraction of E. coli harboring the lsc gene. Expression of lsc was found to be dependent on the vector-based Plac promoter, indicating that the native promoter of lsc was not functional in E. coli. Insertion of an antibiotic resistance cassette in the lsc gene abolished levan synthesis in E. coli. A PCR screening with primers derived from lsc of P. syringae pv. glycinea PG4180 allowed the detection of this gene in a number of related bacteria. PMID- 9726859 TI - Characterization of the bactericidal effects of sodium nitroprusside and other pentacyanonitrosyl complexes on the food spoilage bacterium Clostridium sporogenes. AB - The potent bactericidal activity of sodium nitroprusside (SNP; Na2[Fe(CN)5(NO)]) towards Clostridium sporogenes has been investigated. SNP inhibited cell growth in the concentration range of 10 to 40 microM. Concentrations above 80 microM caused irreversible loss of cell viability and cell lysis. Inhibition of cell growth was similar in complex and in defined media. SNP was found to be unreactive towards individual components of the defined medium, with the exception of cysteine. The chemical characteristics responsible for the potency of SNP were investigated by synthesizing analogs of SNP in which the Fe was replaced by different metals. The inhibitory potency of the pentacyanonitrosyl complexes decreased in the order Fe > Cr > V, which correlates with N-O stretching frequency (vNO). In contrast, the Ru complex which had a vNO comparable to that of Fe was a poor inhibitor. Electron paramagnetic resonance spectroscopy showed that SNP was rapidly reduced to the paramagnetic Fe(I) compound [Fe(CN)4(NO)](2-) on contact with cells. Analysis of fractions from SNP treated cells showed 90% oxidation of thiols in the cell walls compared with those in control cells. The toxicity of SNP involves S-nitrosation and reduction, the lack of toxicity of the Ru analog being consistent with the fact that it has poor reactivity towards thiols. When C. sporogenes cells were exposed to sublethal concentrations of SNP and viewed under the electron microscope, they showed blisters on the surface. These results point to the cell wall surface as a primary point of attack of the nitrosyl complex. PMID- 9726862 TI - In vitro and In vivo characterization of a small-colony variant of the primary form of photorhabdus luminescens MD (Enterobacteriaceae) AB - A small-colony variant (Vsm) of the primary form (Vp) of Photorhabdus luminescens MD from in vitro and in vivo cultures is described. Unlike the primary form, Vp, the Vsm variant is not the preferred diet of its nematode symbiont, a Heterorhabditis sp., does not support development and reproduction of the nematode, and is less pathogenic than Vp to Galleria mellonella larvae. Vsm cells were carried by 25% of infective juveniles, but they comprised a very low percentage ( approximately 0.4%) of the total cells carried by the juvenile. In vitro subculture and in vivo injection into the larvae with either Vp or Vsm always produced a mixture of both Vp and Vsm. In nematode-bacterium-infected G. mellonella larvae, the Vp population in the hemocoel was high (4 x 10(9) to 5 x 10(9) CFU/g of wet insect tissue) at 24 h after infection, decreased about 10 fold by 48 h, and then regained a high level at day 5 before decreasing at day 7 and then remaining relatively constant through day 15 postinfection. The Vsm population, under the same conditions as those of Vp, increased gradually to a high level (9 x 10(8) CFU/g of wet insect tissue) at day 5 postinfection and then declined gradually through day 15. PMID- 9726861 TI - Competitive dominance among strains of luminous bacteria provides an unusual form of evidence for parallel evolution in Sepiolid squid-vibrio symbioses. AB - One of the principal assumptions in symbiosis research is that associated partners have evolved in parallel. We report here experimental evidence for parallel speciation patterns among several partners of the sepiolid squid luminous bacterial symbioses. Molecular phylogenies for 14 species of host squids were derived from sequences of both the nuclear internal transcribed spacer region and the mitochondrial cytochrome oxidase subunit I; the glyceraldehyde phosphate dehydrogenase locus was sequenced for phylogenetic determinations of 7 strains of bacterial symbionts. Comparisons of trees constructed for each of the three loci revealed a parallel phylogeny between the sepiolids and their respective symbionts. Because both the squids and their bacterial partners can be easily cultured independently in the laboratory, we were able to couple these phylogenetic analyses with experiments to examine the ability of the different symbiont strains to compete with each other during the colonization of one of the host species. Our results not only indicate a pronounced dominance of native symbiont strains over nonnative strains, but also reveal a hierarchy of symbiont competency that reflects the phylogenetic relationships of the partners. For the first time, molecular systematics has been coupled with experimental colonization assays to provide evidence for the existence of parallel speciation among a set of animal-bacterial associations. PMID- 9726860 TI - Role of endoproteolytic dibasic proprotein processing in maturation of secretory proteins in Trichoderma reesei. AB - Cell extracts of Trichoderma reesei exhibited dibasic endopeptidase activity toward the carboxylic side of KR, RR, and PR sequences. This activity was stimulated by the presence of Ca2+ ions and localized in vesicles of low bouyant density; it therefore exhibited some similarity to yeast Kex2. Analytical chromatofocusing revealed a single peak of activity. The dibasic endopeptidase activity was strongly and irreversibly inhibited in vitro as well as in vivo by 1 mM p-amidinophenylmethylsulfonyl fluoride (pAPMSF) but not by PMSF at concentrations up to 5 mM. We therefore used pAPMSF to study the role of the dibasic endopeptidase in the secretion of protein by T. reesei. Secretion of xylanase I (proprotein processing sequence -R-R- downward arrow-R- downward arrow A-) and xylanase II (-K-R- downward arrow-Q-) was strongly inhibited by 1 mM pAPMSF, and a larger, unprocessed enzyme form was detected intracellularly under these conditions. Secretion of cellobiohydrolase II (CBH II; -E-R- downward arrow Q-) was only slightly inhibited by pAPMSF, and no accumulation of unprocessed precursors was detected. In contrast, secretion of CBH I (-R-A- downward arrow-Q ) was stimulated by pAPMSF addition, and a simultaneous decrease in the concentration of intracellular CBH I was detected. Similar experiments were also carried out with a single heterologous protein, ShBLE, the phleomycin-binding protein from Streptoalloteichus hindustanus, fused to a series of model proprotein-processing sequences downstream of the expression signals of the Aspergillus nidulans gpdA promoter. Consistent with the results obtained with homologous proteins, pAPMSF inhibited the secretion of ShBLE with fusions containing dibasic (RK and KR) target sequences, but it even stimulated secretion in fusions to LR, NHA, and EHA target sequences. Addition of 5 mM PMSF, a nonspecific inhibitor of serine protease, nonspecifically inhibited the secretion of heterologous proteins from fusions bearing the NHA and LR targets. These data point to the existence of different endoproteolytic proprotein processing enzymes in T. reesei and demonstrate that dibasic processing is obligatory for the secretion of the proproteins containing this target. PMID- 9726863 TI - Comparative study of pressure-induced germination of Bacillus subtilis spores at low and high pressures. AB - We have studied pressure-induced germination of Bacillus subtilis spores at moderate (100 MPa) and high (500 to 600 MPa) pressures. Although we found comparable germination efficiencies under both conditions by using heat sensitivity as a criterion for germination, the sensitivity of pressure germinated spores to some other agents was found to depend on the pressure used. Spores germinated at 100 MPa were more sensitive to pressure (>200 MPa), UV light, and hydrogen peroxide than were those germinated at 600 MPa. Since small, acid-soluble proteins (SASPs) and dipicolinic acid (DPA) are known to be involved in spore resistance to UV light and hydrogen peroxide, we studied the fate of these compounds during pressure germination. DPA was released upon both low- and high-pressure germination, but SASP degradation, which normally accompanies nutrient-induced germination, occurred upon low-pressure germination but not upon high-pressure germination. These results adequately explain the UV and hydrogen peroxide resistance of spores germinated at 600 MPa. The resistance to pressure inactivation of 600-MPa-germinated spores could also, at least partly, be attributed to alpha/beta-type SASPs, since mutants deficient in alpha/beta-type SASPs were more sensitive to inactivation at 600 MPa. Further, germination at 100 MPa resulted in rapid ATP generation, as is the case in nutrient-induced germination, but no ATP was formed during germination at 600 MPa. These results suggest that spore germination can be initiated by low- and high-pressure treatments but is arrested at an early stage in the latter case. The implications for the use of high pressure as a preservation treatment are discussed. PMID- 9726864 TI - Biotransformation of the major fungal metabolite 3,5-dichloro- p-anisyl alcohol under anaerobic conditions and its role in formation of Bis(3,5-dichloro-4 Hydroxyphenyl)methane. AB - Higher fungi have a widespread capacity for biosynthesis of organohalogens. Commonly occurring chloroaromatic fungal metabolites can end up in anaerobic microniches at the boundary of fungal colonies and wetland soils. The aim of this study was to investigate the environmental fate of a major fungal metabolite, 3, 5-dichloro-p-anisyl alcohol, under anaerobic conditions. This compound was incubated with methanogenic sludge to study its biotransformation reactions. Initially, 3,5-dichloro-p-anisyl alcohol was readily demethylated in stoichiometric quantities to 3, 5-dichloro-4-hydroxybenzyl alcohol. The demethylated product was converted further via two routes: a biotic route leading to the formation of 3,5-dichloro-4-hydroxybenzoate and 2,6-dichlorophenol, as well as an abiotic route leading to the formation of bis(3, 5-dichloro-4 hydroxyphenyl)methane. In the first route, the benzyl alcohol moiety on the aromatic ring was oxidized, giving 3, 5-dichloro-4-hydroxybenzoate as a transient or accumulating product, depending on the type of methanogenic sludge used. In sludge previously adapted to low-molecular-weight lignin from straw, a part of the 3,5-dichloro-4-hydroxybenzoate was decarboxylated, yielding detectable levels of 2,6-dichlorophenol. In the second route, 3, 5-dichloro-4-hydroxybenzyl alcohol dimerized, leading to the formation of a tetrachlorinated bisphenolic compound, which was identified as bis(3,5-dichloro-4-hydroxyphenyl)methane. Since formation of this dimer was also observed in incubations with autoclaved sludge spiked with 3,5-dichloro-4-hydroxybenzyl alcohol, it was concluded that its formation was due to an abiotic process. However, demethylation of the fungal metabolite by biological processes was a prerequisite for dimerization. The most probable reaction mechanism leading to the formation of the tetrachlorinated dimer in the absence of oxygen is presented, and the possible environmental implications of its natural occurrence are discussed. PMID- 9726865 TI - Role of the regulatory gene areA of Aspergillus oryzae in nitrogen metabolism. AB - The areA gene of Aspergillus oryzae was cloned by cross-hybridization with the Aspergillus nidulans areA gene and was found to encode an 866-amino-acid protein that is very similar to other fungal nitrogen regulatory proteins. The A. oryzae areA gene can complement A. nidulans areA loss-of-function mutations. Functional analyses indicated that the N-terminal region of the A. oryzae AreA protein was dispensable for function and revealed a probable acidic activation domain in the protein. C-terminal truncation of the protein resulted in derepression of several nitrogen-controlled activities in A. nidulans, while deletions extending into the conserved GATA type zinc finger region abolished the activator function. The A. oryzae areA gene was inactivated by replacement with the A. oryzae pyrG gene. Strains containing the resulting areA deletion grew as well as the wild-type strain on glutamine but were unable to grow vigorously on other nitrogen sources, including ammonium. While A. oryzae exhibited reduced growth on 10 mM ammonium, the results of growth tests indicated that areA mutants of both A. oryzae and A. nidulans were affected in utilization of low concentrations of ammonium. The levels of the major nitrogen assimilatory enzymes, NADP-linked glutamate dehydrogenase (EC 1.4.1.4) and glutamine synthetase (EC 6.3.1.2), were determined. In both A. oryzae and A. nidulans areA mutants, the NADP-glutamate dehydrogenase levels were reduced, whereas the glutamine synthetase levels were not affected. These results suggest that the AreA protein may play an important role in the regulation of nitrogen assimilation in addition to its previously established regulatory role in nitrogen catabolism. PMID- 9726866 TI - Nucleic acid (DNA, RNA) quantification and RNA/DNA ratio determination in marine sediments: comparison of spectrophotometric, fluorometric, and HighPerformance liquid chromatography methods and estimation of detrital DNA AB - In this study, we compared three methods for extraction and quantification of RNA and DNA from marine sediments: (i) a spectrophotometric method using the diphenylamine assay; (ii) a fluorometric method utilizing selective fluorochromes (thiazole orange for total nucleic acids and Hoechst 33258 for DNA); and (iii) a high-pressure liquid chromatography (HPLC) method which uses a specific column to separate RNA and DNA and UV absorption of the nucleic acids for quantification. Sediment samples were collected in the oligotrophic Cretan Sea (eastern Mediterranean, from 40 to 1,540 m in depth) and compared to the distribution and composition of the benthic microbial assemblages (i.e., bacteria and microprotozoa). DNA concentrations measured spectrophotometrically and by HPLC were not significantly different, while fluorometric yields were significantly lower. Such differences appear mainly due to fact that the stain-DNA complex is strongly dependent on the DNA composition and structure. RNA concentrations determined by the three methods displayed some differences; fluorometric and spectrophotometric methods obtain RNA concentration by difference and therefore may be biased by DNA estimates. By contrast, the HPLC method provides independent assessments of RNA and DNA concentrations. We tentatively estimated the contribution of the detrital DNA to the total DNA pools in two ways. The two calculations provided quite similar results indicating that the majority of the DNA pool in the deep-sea sediments was detrital. Microbial RNA generally accounted for almost the entire sedimentary RNA pools below 100-m depth. RNA concentrations were found to decrease along the Cretan shelf and slope. The RNA/DNA ratio calculated by using fluorometric DNA concentrations was significantly correlated with values of sediment community oxygen consumption only below 100-m depth (dominated by the microbial biomass). These data suggest that the RNA/DNA ratio, based on fluorometric estimates of DNA, can be used as an indicator of benthic metabolic activity, but only when metazoan contribution to the microbial DNA is negligible. PMID- 9726867 TI - Rapid determination of bacterial abundance, biovolume, morphology, and growth by neural network-based image analysis AB - Annual bacterial plankton dynamics at several depths and locations in the Baltic Sea were studied by image analysis. Individual bacteria were classified by using an artificial neural network which also effectively identified nonbacterial objects. Cell counts and frequencies of dividing cells were determined, and the data obtained agreed well with visual observations and previously published values. Cell volumes were measured accurately by comparison with bead standards. The survey included 690 images from a total of 138 samples. Each image contained approximately 200 bacteria. The images were analyzed automatically at a rate of 100 images per h. Bacterial abundance exhibited coherent patterns with time and depth, and there were distinct subsurface peaks in the summer months. Four distinct morphological classes were resolved by the image analyzer, and the dynamics of each could be visualized. The bacterial growth rates estimated from frequencies of dividing cells were different from the bacterial growth rates estimated by the thymidine incorporation method. With minor modifications, the image analysis technique described here can be used to analyze other planktonic classes. PMID- 9726868 TI - Contribution of indole-3-acetic acid production to the epiphytic fitness of erwinia herbicola AB - Erwinia herbicola 299R produces large quantities of indole-3-acetic acid (IAA) in culture media supplemented with L-tryptophan. To assess the contribution of IAA production to epiphytic fitness, the population dynamics of the wild-type strain and an IAA-deficient mutant of this strain on leaves were studied. Strain 299XYLE, an isogenic IAA-deficient mutant of strain 299R, was constructed by insertional interruption of the indolepyruvate decarboxylase gene of strain 299R with the xylE gene, which encodes a 2,3-catechol dioxygenase from Pseudomonas putida mt-2. The xylE gene provided a useful marker for monitoring populations of the IAA-deficient mutant strain in mixed populations with the parental strain in ecological studies. A root bioassay for IAA, in which strain 299XYLE inhibited significantly less root elongation than strain 299R, provided evidence that E. herbicola produces IAA on plant surfaces in amounts sufficient to affect the physiology of its host and that IAA production in strain 299R is not solely an in vitro phenomenon. The epiphytic fitness of strains 299R and 299XYLE was evaluated in greenhouse and field studies by analysis of changes in the ratio of the population sizes of these two strains after inoculation as mixtures onto plants. Populations of the parental strain increased to approximately twice those of the IAA-deficient mutant strain after coinoculation in a proportion of 1:1 onto bean plants in the greenhouse and onto pear flowers in field studies. In all experiments, the ratio of the population sizes of strain 299R and 299XYLE increased during periods of active growth on plant tissue but not when population sizes were not increasing with time. PMID- 9726869 TI - Biosensor determination of the microscale distribution of nitrate, nitrate assimilation, nitrification, and denitrification in a diatom-inhabited freshwater sediment AB - High-resolution NO3- profiles in freshwater sediment covered with benthic diatoms were obtained with a new microscale NO3- biosensor characterized by absence of interference from chemical species other than NO2- and N2O. Analysis of the microprofiles obtained indicated no nitrification during darkness, high rates of nitrification and a tight coupling between nitrification and denitrification during illumination, and substantial rates of NO3- assimilation during illumination. Nitrification during darkness could be induced by purging the bulk water with O2 gas, indicating that the stimulatory effect on nitrification by illumination was caused by algal production of O2. NH4+ addition did not stimulate nitrification during darkness when O2 was restricted to the upper 1-mm layer, and there was thus a low nitrification potential in the permanently oxic top 1 mm of the sediment. PMID- 9726870 TI - Initial transformations in the biodegradation of benzothiazoles by Rhodococcus isolates. AB - Benzothiazole-2-sulfonate (BTSO3) is one of the side products occurring in 2 mercaptobenzothiazole (MBT) production wastewater. We are the first to isolate an axenic culture capable of BTSO3 degradation. The isolate was identified as a Rhodococcus erythropolis strain and also degraded 2-hydroxybenzothiazole (OBT) and benzothiazole (BT), but not MBT, which was found to inhibit the biodegradation of OBT, BT, and BTSO3. In anaerobic resting cell assays, BTSO3 was transformed into OBT in stoichiometric amounts. Under aerobic conditions, OBT was observed as an intermediate in BT breakdown and an unknown compound transiently accumulated in several assays. This product was identified as a dihydroxybenzothiazole. Benzothiazole degradation pathways seem to converge into OBT, which is then transformed further into the dihydroxy derivative. PMID- 9726871 TI - Comparative studies of class IIa bacteriocins of lactic acid bacteria. AB - Four class IIa bacteriocins (pediocin PA-1, enterocin A, sakacin P, and curvacin A) were purified to homogeneity and tested for activity toward a variety of indicator strains. Pediocin PA-1 and enterocin A inhibited more strains and had generally lower MICs than sakacin P and curvacin A. The antagonistic activity of pediocin-PA1 and enterocin A was much more sensitive to reduction of disulfide bonds than the antagonistic activity of sakacin P and curvacin A, suggesting that an extra disulfide bond that is present in the former two may contribute to their high levels of activity. The food pathogen Listeria monocytogenes was among the most sensitive indicator strains for all four bacteriocins. Enterocin A was most effective in inhibiting Listeria, having MICs in the range of 0.1 to 1 ng/ml. Sakacin P had the interesting property of being very active toward Listeria but not having concomitant high levels of activity toward lactic acid bacteria. Strains producing class IIa bacteriocins displayed various degrees of resistance toward noncognate class IIa bacteriocins; for the sakacin P producer, it was shown that this resistance is correlated with the expression of immunity genes. It is hypothesized that variation in the presence and/or expression of such immunity genes accounts in part for the remarkably large variation in bacteriocin sensitivity displayed by lactic acid bacteria. PMID- 9726872 TI - Enzymatic properties of a novel liquefying alpha-amylase from an alkaliphilic Bacillus isolate and entire nucleotide and amino acid sequences. AB - A novel liquefying alpha-amylase (LAMY) was found in cultures of an alkaliphilic Bacillus isolate, KSM-1378. The specific activity of purified LAMY was approximately 5,000 U mg of protein-1, a value two- to fivefold greater between pH 5 and 10 than that of an industrial, thermostable Bacillus licheniformis enzyme. The enzyme had a pH optimum of 8.0 to 8.5 and displayed maximum activity at 55 degreesC. The molecular mass deduced from sodium dodecyl sulfate polyacrylamide gel electrophoresis was approximately 53 kDa, and the apparent isoelectric point was around pH 9. This enzyme efficiently hydrolyzed various carbohydrates to yield maltotriose, maltopentaose, maltohexaose, and maltose as major end products after completion of the reaction. Maltooligosaccharides in the maltose-to-maltopentaose range were unhydrolyzable by the enzyme. The structural gene for LAMY contained a single open reading frame 1, 548 bp in length, corresponding to 516 amino acids that included a signal peptide of 31 amino acids. The calculated molecular mass of the extracellular mature enzyme was 55,391 Da. LAMY exhibited relatively low amino acid identity to other liquefying amylases, such as the enzymes from B. licheniformis (68.9%), Bacillus amyloliquefaciens (66.7%), and Bacillus stearothermophilus (68.6%). The four conserved regions, designated I, II, III, and IV, and the putative catalytic triad were found in the deduced amino acid sequence of LAMY. Essentially, the sequence of LAMY was consistent with the tertiary structures of reported amylolytic enzymes, which are composed of domains A, B, and C and which include the well-known (alpha/beta)8 barrel motif in domain A. PMID- 9726874 TI - Aerobic growth on nitroglycerin as the sole carbon, nitrogen, and energy source by a mixed bacterial culture. AB - Nitroglycerin (glycerol trinitrate [GTN]), an explosive and vasodilatory compound, was metabolized by mixed microbial cultures from aeration tank sludge previously exposed to GTN. Aerobic enrichment cultures removed GTN rapidly in the absence of a supplemental carbon source. Complete denitration of GTN, provided as the sole C and N source, was observed in aerobic batch cultures and proceeded stepwise via the dinitrate and mononitrate isomers, with successive steps occurring at lower rates. The denitration of all glycerol nitrate esters was found to be concomitant, and 1, 2-glycerol dinitrate (1,2-GDN) and 2-glycerol mononitrate (2-GMN) were the primary GDN and GMN isomers observed. Denitration of GTN resulted in release of primarily nitrite-N, indicating a reductive denitration mechanism. Biomass growth at the expense of GTN was verified by optical density and plate count measurements. The kinetics of GTN biotransformation were 10-fold faster than reported for complete GTN denitration under anaerobic conditions. A maximum specific growth rate of 0.048 +/- 0.005 h-1 (mean +/- standard deviation) was estimated for the mixed culture at 25 degreesC. Evidence of GTN toxicity was observed at GTN concentrations above 0. 3 mM. To our knowledge, this is the first report of complete denitration of GTN used as a primary growth substrate by a bacterial culture under aerobic conditions. PMID- 9726875 TI - A new intermediate in the mineralization of 3,4-dichloroaniline by the white rot fungus Phanerochaete chrysosporium. AB - Phanerochaete chrysosporium ATCC 34541 has been reported to be unable to mineralize 3,4-dichloroaniline (DCA). However, high mineralization is now shown to occur when a fermentation temperature of 37 degrees and gassing with oxygen are used. Mineralization did not correlate with lignin peroxidase activity. The latter was high under C limitation and low under N limitation, whereas the reverse was true for mineralization. The kinetics of DCA metabolism was studied in low-N and low-C and C- and N-rich culture media by metabolite analysis and 14CO2 determination. In all cases, DCA disappeared within 2 days, and a novel highly polar conjugate termed DCAX accumulated in the growth medium. This metabolite was a dead-end product under C and N enrichment. In oxygenated low-C medium and in much higher yield in oxygenated low-N medium, DCAX was converted to DCA-succinimide and then mineralized. DCAX was purified by high-performance liquid chromatography and identified as N-(3,4-dichlorophenyl)-alpha-ketoglutaryl delta-amide by high-performance liquid chromatography and mass spectroscopy, gas chromatography and mass spectroscopy, and nuclear magnetic resonance spectroscopy. The formation of conjugate intermediates is proposed to facilitate mineralization because the sensitive amino group of DCA needs protection so that ring cleavage rather than oligomerization can occur. PMID- 9726873 TI - Substrate specificity of and product formation by muconate cycloisomerases: an analysis of wild-type enzymes and engineered variants. AB - Muconate cycloisomerases play a crucial role in the bacterial degradation of aromatic compounds by converting cis,cis-muconate, the product of catechol ring cleavage, to (4S)-muconolactone. Chloromuconate cycloisomerases catalyze both the corresponding reaction and a dehalogenation reaction in the transformation of chloroaromatic compounds. This study reports the first thorough examination of the substrate specificity of the muconate cycloisomerases from Pseudomonas putida PRS2000 and Acinetobacter "calcoaceticus" ADP1. We show that they transform, in addition to cis,cis-muconate, 3-fluoro-, 2-methyl-, and 3-methyl-cis, cis muconate with high specificity constants but not 2-fluoro-, 2-chloro-, 3-chloro-, or 2,4-dichloro-cis,cis-muconate. Based on known three-dimensional structures, variants of P. putida muconate cycloisomerase were constructed by site-directed mutagenesis to contain amino acids found in equivalent positions in chloromuconate cycloisomerases. Some of the variants had significantly increased specificity constants for 3-chloro- or 2,4-dichloromuconate (e.g., A271S and I54V showed 27- and 22-fold increases, respectively, for the former substrate). These kinetic improvements were not accompanied by a change from protoanemonin to cis,cis-dienelactone as the product of 3-chloro-cis,cis-muconate conversion. The rate of 2-chloro-cis,cis-muconate turnover was not significantly improved, nor was this compound dehalogenated to any significant extent. However, the direction of 2-chloro-cis,cis-muconate cycloisomerization could be influenced by amino acid exchange. While the wild-type enzyme discriminated only slightly between the two possible cycloisomerization directions, some of the enzyme variants showed a strong preference for either (+)-2-chloro- or (+)-5-chloromuconolactone formation. These results show that the different catalytic characteristics of muconate and chloromuconate cycloisomerases are due to a number of features that can be changed independently of each other. PMID- 9726877 TI - Conversion of methionine to thiols by lactococci, lactobacilli, and brevibacteria AB - Methanethiol has been strongly associated with desirable Cheddar cheese flavor and can be formed from the degradation of methionine (Met) via a number of microbial enzymes. Methionine gamma-lyase is thought to play a major role in the catabolism of Met and generation of methanethiol in several species of bacteria. Other enzymes that have been reported to be capable of producing methanethiol from Met in lactic acid bacteria include cystathionine beta-lyase and cystathionine gamma-lyase. The objective of this study was to determine the production, stability, and activities of the enzymes involved in methanethiol generation in bacteria associated with cheese making. Lactococci and lactobacilli were observed to contain high levels of enzymes that acted primarily on cystathionine. Enzyme activity was dependent on the concentration of sulfur amino acids in the growth medium. Met aminotransferase activity was detected in all of the lactic acid bacteria tested and alpha-ketoglutarate was used as the amino group acceptor. In Lactococcus lactis subsp. cremoris S2, Met aminotransferase was repressed with increasing concentrations of Met in the growth medium. While no Met aminotransferase activity was detected in Brevibacterium linens BL2, it possessed high levels of L-methionine gamma-lyase that was induced by addition of Met to the growth medium. Met demethiolation activity at pH 5.2 with 4% NaCl was not detected in cell extracts but was detected in whole cells. These data suggest that Met degradation in Cheddar cheese will depend on the organism used in production, the amount of enzyme released during aging, and the amount of Met in the matrix. PMID- 9726876 TI - Identification and characterization of Leuconostoc carnosum, associated with production and spoilage of vacuum-packaged, sliced, cooked ham. AB - Leuconostoc carnosum was shown to be the specific spoilage organism in vacuum packaged, sliced, cooked ham showing spoilage during 3 weeks of shelf life. Identification of the specific spoilage organism was done by use of phenotypic data and ClaI, EcoRI, and HindIII reference strain ribopatterns. One hundred L. carnosum isolates associated with the production and spoilage of the ham were further characterized by pulsed-field gel electrophoresis (PFGE), together with some meat-associated Leuconostoc species: L. citreum, L. gelidum, L. mesenteroides subsp. dextranicum, and L. mesenteroides subsp. mesenteroides. ApaI and SmaI digests divided the industrial L. carnosum strains into 25 different PFGE types, ApaI and SmaI types being consistent. Only one specific PFGE type was associated with the spoiled packages. This type also was detected in air and raw meat mass samples. The spoilage strain did not produce bacteriocins. Only seven isolates belonging to three different PFGE types produced bacteriocins. Similarity analysis of the industrial L. carnosum strains revealed a homogeneous cluster which could be divided into eight subclusters consisting of strains having at most three-fragment differences. The L. carnosum cluster was clearly distinguished from the other meat-associated leuconostoc clusters, with the exception of the L. carnosum type strain. Ribotyping can be very helpful in the identification of L. carnosum, but its discriminatory power is too weak for strain characterization. PFGE provides good discrimination for studies dealing with the properties of homogeneous L. carnosum strains. PMID- 9726878 TI - Purification and characterization of L-methionine gamma-lyase from brevibacterium linens BL2 AB - L-Methionine gamma-lyase (EC 4.4.1.11) was purified to homogeneity from Brevibacterium linens BL2, a coryneform bacterium which has been used successfully as an adjunct bacterium to improve the flavor of Cheddar cheese. The enzyme catalyzes the alpha,gamma elimination of methionine to produce methanethiol, alpha-ketobutyrate, and ammonia. It is a pyridoxal phosphate dependent enzyme, with a native molecular mass of approximately 170 kDa, consisting of four identical subunits of 43 kDa each. The purified enzyme had optimum activity at pH 7.5 and was stable at pHs ranging from 6.0 to 8.0 for 24 h. The pure enzyme had its highest activity at 25 degreesC but was active between 5 and 50 degreesC. Activity was inhibited by carbonyl reagents, completely inactivated by DL-propargylglycine, and unaffected by metal-chelating agents. The pure enzyme had catalytic properties similar to those of L-methionine gamma-lyase from Pseudomonas putida. Its Km for the catalysis of methionine was 6.12 mM, and its maximum rate of catalysis was 7.0 &mgr;mol min-1 mg-1. The enzyme was active under salt and pH conditions found in ripening Cheddar cheese but susceptible to degradation by intracellular proteases. PMID- 9726879 TI - Confirmation of the human-pathogenic microsporidia Enterocytozoon bieneusi, Encephalitozoon intestinalis, and Vittaforma corneae in water. AB - Microsporidia, as a group, cause a wide range of infections, though two species of microsporidia in particular, Enterocytozoon bieneusi and Encephalitozoon intestinalis, are associated with gastrointestinal disease in humans. To date, the mode of transmission and environmental occurrence of microsporidia have not been elucidated due to lack of sensitive and specific screening methods. The present study was undertaken with recently developed methods to screen several significant water sources. Water concentrates were subjected to community DNA extraction followed by microsporidium-specific PCR amplification, PCR sequencing, and database homology comparison. A total of 14 water concentrates were screened; 7 of these contained human-pathogenic microsporidia. The presence of Encephalitozoon intestinalis was confirmed in tertiary sewage effluent, surface water, and groundwater; the presence of Enterocytozoon bieneusi was confirmed in surface water; and the presence of Vittaforma corneae was confirmed in tertiary effluent. Thus, this study represents the first confirmation, to the species level, of human-pathogenic microsporidia in water, indicating that these human pathogenic microsporidia may be waterborne pathogens. PMID- 9726880 TI - Variations of bacterial populations in human feces measured by fluorescent in situ hybridization with group-specific 16S rRNA-targeted oligonucleotide probes. AB - Six 16S rRNA-targeted oligonucleotide probes were designed, validated, and used to quantify predominant groups of anaerobic bacteria in human fecal samples. A set of two probes was specific for species of the Bacteroides fragilis group and the species Bacteroides distasonis. Two others were designed to detect species of the Clostridium histolyticum and the Clostridium lituseburense groups. Another probe was designed for the genera Streptococcus and Lactococcus, and the final probe was designed for the species of the Clostridium coccoides-Eubacterium rectale group. The temperature of dissociation of each of the probes was determined. The specificities of the probes for a collection of target and reference organisms were tested by dot blot hybridization and fluorescent in situ hybridization (FISH). The new probes were used in initial FISH experiments to enumerate human fecal bacteria. The combination of the two Bacteroides-specific probes detected a mean of 5.4 x 10(10) cells per g (dry weight) of feces; the Clostridium coccoides-Eubacterium rectale group-specific probe detected a mean of 7.2 x 10(10) cells per g (dry weight) of feces. The Clostridium histolyticum, Clostridium lituseburense, and Streptococcus-Lactococcus group-specific probes detected only numbers of cells ranging from 1 x 10(7) to 7 x 10(8) per g (dry weight) of feces. Three of the newly designed probes and three additional probes were used in further FISH experiments to study the fecal flora composition of nine volunteers over a period of 8 months. The combination of probes was able to detect at least two-thirds of the fecal flora. The normal biological variations within the fecal populations of the volunteers were determined and indicated that these variations should be considered when evaluating the effects of agents modulating the flora. PMID- 9726881 TI - Growth rate regulation of rRNA content of a marine synechococcus (Cyanobacterium) strain AB - The relationship between growth rate and rRNA content in a marine Synechococcus strain was examined. A combination of flow cytometry and whole-cell hybridization with fluorescently labeled 16S rRNA-targeted oligonucleotide probes was used to measure the rRNA content of Synechococcus strain WH8101 cells grown at a range of light-limited growth rates. The sensitivity of this approach was sufficient for the analysis of rRNA even in very slowly growing Synechococcus cells (&mgr; = 0.15 day-1). The relationship between growth rate and cellular rRNA content comprised three phases: (i) at low growth rates (< approximately 0.7 day-1), rRNA cell-1 remained approximately constant; (ii) at intermediate rates ( approximately 0. 7 - 1.6 day-1), rRNA cell-1 increased proportionally with growth rate; and (iii) at the highest, light-saturated rates (> approximately 1.6 day 1), rRNA cell-1 dropped abruptly. Total cellular RNA (as measured with the nucleic acid stain SYBR Green II) was well correlated with the probe-based measure of rRNA and varied in a similar manner with growth rate. Mean cell volume and rRNA concentration (amount of rRNA per cubic micrometer) were related to growth rate in a manner similar to rRNA cell-1, although the overall magnitude of change in both cases was reduced. These patterns are hypothesized to reflect an approximately linear increase in ribosome efficiency with increasing growth rate, which is consistent with the prevailing prokaryotic model at low growth rates. Taken together, these results support the notion that measurements of cellular rRNA content might be useful for estimating in situ growth rates in natural Synechococcus populations. PMID- 9726882 TI - Direct determination of carbon and nitrogen contents of natural bacterial assemblages in marine environments AB - In order to better estimate bacterial biomass in marine environments, we developed a novel technique for direct measurement of carbon and nitrogen contents of natural bacterial assemblages. Bacterial cells were separated from phytoplankton and detritus with glass fiber and membrane filters (pore size, 0.8 &mgr;m) and then concentrated by tangential flow filtration. The concentrate was used for the determination of amounts of organic carbon and nitrogen by a high temperature catalytic oxidation method, and after it was stained with 4',6 diamidino-2-phenylindole, cell abundance was determined by epifluorescence microscopy. We found that the average contents of carbon and nitrogen for oceanic bacterial assemblages were 12.4 +/- 6.3 and 2.1 +/- 1.1 fg cell-1 (mean +/- standard deviation; n = 6), respectively. Corresponding values for coastal bacterial assemblages were 30.2 +/- 12.3 fg of C cell-1 and 5.8 +/- 1.5 fg of N cell-1 (n = 5), significantly higher than those for oceanic bacteria (two-tailed Student's t test; P < 0.03). There was no significant difference (P > 0.2) in the bacterial C:N ratio (atom atom-1) between oceanic (6.8 +/- 1.2) and coastal (5.9 +/- 1.1) assemblages. Our estimates support the previous proposition that bacteria contribute substantially to total biomass in marine environments, but they also suggest that the use of a single conversion factor for diverse marine environments can lead to large errors in assessing the role of bacteria in food webs and biogeochemical cycles. The use of a factor, 20 fg of C cell-1, which has been widely adopted in recent studies may result in the overestimation (by as much as 330%) of bacterial biomass in open oceans and in the underestimation (by as much as 40%) of bacterial biomass in coastal environments. PMID- 9726883 TI - Characterization of a defined 2,3,5, 6-tetrachlorobiphenyl-ortho-dechlorinating microbial community by comparative sequence analysis of genes coding for 16S rRNA. AB - Defined microbial communities were developed by combining selective enrichment with molecular monitoring of total community genes coding for 16S rRNAs (16S rDNAs) to identify potential polychlorinated biphenyl (PCB)-dechlorinating anaerobes that ortho dechlorinate 2,3, 5,6-tetrachlorobiphenyl. In enrichment cultures that contained a defined estuarine medium, three fatty acids, and sterile sediment, a Clostridium sp. was predominant in the absence of added PCB, but undescribed species in the delta subgroup of the class Proteobacteria, the low-G+C gram-positive subgroup, the Thermotogales subgroup, and a single species with sequence similarity to the deeply branching species Dehalococcoides ethenogenes were more predominant during active dechlorination of the PCB. Species with high sequence similarities to Methanomicrobiales and Methanosarcinales archaeal subgroups were predominant in both dechlorinating and nondechlorinating enrichment cultures. Deletion of sediment from PCB dechlorinating enrichment cultures reduced the rate of dechlorination and the diversity of the community. Substitution of sodium acetate for the mixture of three fatty acids increased the rate of dechlorination, further reduced the community diversity, and caused a shift in the predominant species that included restriction fragment length polymorphism patterns not previously detected. Although PCB-dechlorinating cultures were methanogenic, inhibition of methanogenesis and elimination of the archaeal community by addition of bromoethanesulfonic acid only slightly inhibited dechlorination, indicating that the archaea were not required for ortho dechlorination of the congener. Deletion of Clostridium spp. from the community profile by addition of vancomycin only slightly reduced dechlorination. However, addition of sodium molybdate, an inhibitor of sulfate reduction, inhibited dechlorination and deleted selected species from the community profiles of the class Bacteria. With the exception of one 16S rDNA sequence that had the highest sequence similarity to the obligate perchloroethylene-dechlorinating Dehalococcoides, the 16S rDNA sequences associated with PCB ortho dechlorination had high sequence similarities to the delta, low-G+C gram-positive, and Thermotogales subgroups, which all include sulfur-, sulfate-, and/or iron(III)-respiring bacterial species. PMID- 9726884 TI - Biodegradation of atrazine by Agrobacterium radiobacter J14a and use of this strain in bioremediation of contaminated soil. AB - We examined the ability of a soil bacterium, Agrobacterium radiobacter J14a, to degrade the herbicide atrazine under a variety of cultural conditions, and we used this bacterium to increase the biodegradation of atrazine in soils from agricultural chemical distribution sites. J14a cells grown in nitrogen-free medium with citrate and sucrose as carbon sources mineralized 94% of 50 microgram of [14C-U-ring]atrazine ml-1 in 72 h with a concurrent increase in the population size from 7.9 x 10(5) to 5.0 x 10(7) cells ml-1. Under these conditions cells mineralized the [ethyl-14C]atrazine and incorporated approximately 30% of the 14C into the J14a biomass. Cells grown in medium without additional carbon and nitrogen sources degraded atrazine, but the cell numbers did not increase. Metabolites produced by J14a during atrazine degradation include hydroxyatrazine, deethylatrazine, and deethyl-hydroxyatrazine. The addition of 10(5) J14a cells g 1 into soil with a low indigenous population of atrazine degraders treated with 50 and 200 microgram of atrazine g-1 soil resulted in two to five times higher mineralization than in the noninoculated soil. Sucrose addition did not result in significantly faster mineralization rates or shorten degradation lag times. However, J14a introduction (10(5) cells g-1) into another soil with a larger indigenous atrazine-mineralizing population reduced the atrazine degradation lag times below those in noninoculated treatments but did not generally increase total atrazine mineralization. PMID- 9726885 TI - Viral pollution in the environment and in shellfish: human adenovirus detection by PCR as an index of human viruses. AB - A study of the presence of human viruses (adenoviruses, enteroviruses, and hepatitis A viruses [HAVs]) in environmental and shellfish samples was carried out by applying DNA and cDNA amplification techniques by PCR. The detection of human adenoviruses by PCR was also examined as a potential molecular test to monitor viral pollution. The samples studied were urban and slaughterhouse sewage, river water, seawater, and shellfish. Enteroviruses were quantified by PFU in Buffalo green monkey kidney cells and fecal coliforms and phages of Bacteroides fragilis HSP40 were also evaluated in some of the samples. The amplification of viral DNA and cDNA has shown a high prevalence of human viruses that would not be detected by the use of classical techniques, such as the quantification of PFU in cell lines. The results of the analysis of slaughterhouse sewage samples together with the test of farm animal feces indicate that the adenoviruses and the HAVs detected in the environment are mostly of human origin. A significative correlation between the detection of human viruses by PCR and the values of bacteriophages of B. fragilis HSP40 in urban raw sewage was observed. Human adenoviruses were the viruses most frequently detected throughout the year, and all the samples that were positive for enteroviruses or HAVs were also positive for human adenoviruses. The results suggest that the detection of adenoviruses by PCR could be used as an index of the presence of human viruses in the environment where a molecular index is acceptable. PMID- 9726886 TI - Mercury resistance is encoded by transferable giant linear plasmids in two chesapeake bay Streptomyces strains. AB - The Streptomyces strains CHR3 and CHR28, isolated from the Baltimore Inner Harbor, contained two and one, respectively, giant linear plasmids which carry terminally bound proteins. The plasmids pRJ3L (322 kb), from CHR3, and pRJ28 (330 kb), from CHR28, carry genes homologous to the previously characterized chromosomal Streptomyces lividans 66 operon encoding resistance against mercuric compounds. Both plasmids are transmissible (without any detectable rearrangement) to the chloramphenicol-resistant S. lividans TK24 strain lacking plasmids and carrying a chromosomal deletion of the mer operon. S. lividans TK24 conjugants harboring pRJ3L or pRJ28 exhibited profiles of mercury resistance to mercuric compounds similar to those of Streptomyces strains CHR3 and CHR28. PMID- 9726887 TI - PCR-based DNA amplification and presumptive detection of Escherichia coli O157:H7 with an internal fluorogenic probe and the 5' nuclease (TaqMan) assay. AB - Presumptive identification of Escherichia coli O157:H7 is possible in an individual, nonmultiplexed PCR if the reaction targets the enterohemorrhagic E. coli (EHEC) eaeA gene. In this report, we describe the development and evaluation of the sensitivity and specificity of a PCR-based 5' nuclease assay for presumptively detecting E. coli O157:H7 DNA. The specificity of the eaeA-based 5' nuclease assay system was sufficient to correctly identify all E. coli O157:H7 strains evaluated, mirroring the previously described specificity of the PCR primers. The SZ-primed, eaeA-targeted 5' nuclease detection assay was capable of rapid, semiautomated, presumptive detection of E. coli O157:H7 when >/=10(3) CFU/ml was present in modified tryptic soy broth (mTSB) or modified E. coli broth and when >/=10(4) CFU/ml was present in ground beef-mTSB mixtures. Incorporating an immunomagnetic separation (IMS) step, followed by a secondary enrichment culturing step and DNA recovery with a QIAamp tissue kit (Qiagen), improved the detection threshold to >/=10(2) CFU/ml. Surprisingly, immediately after IMS, the sensitivity of culturing on sorbitol MacConkey agar containing cefeximine and tellurite (CT-SMAC) was such that identifiable colonies were demonstrated only when >/=10(4) CFU/ml was present in the sample. Several factors that might be involved in creating these false-negative CT-SMAC culture results are discussed. The SZ-primed, eaeA-targeted 5' nuclease detection system demonstrated that it can be integrated readily into standard culturing procedures and that the assay can be useful as a rapid, automatable process for the presumptive identification of E. coli O157:H7 in ground beef and potentially in other food and environmental samples. PMID- 9726888 TI - Purification and characterization of three thermostable endochitinases of a noble Bacillus strain, MH-1, isolated from chitin-containing compost. AB - A thermophilic and actinic bacterium strain, MH-1, which produced three different endochitinases in its culture fluid was isolated from chitin-containing compost. The microorganism did not grow in any of the usual media for actinomyces but only in colloidal chitin supplemented with yeast extract and (2, 6-O-dimethyl)-beta cyclodextrin. Compost extract enhanced its growth. In spite of the formation of branched mycelia, other properties of the strain, such as the formation of endospores, the presence of meso-diaminopimelic acid in the cell wall, the percent G+C of DNA (55%), and the partial 16S ribosomal DNA sequence, indicated that strain MH-1 should belong to the genus Bacillus. Three isoforms of endochitinase (L, M, and S) were purified to homogeneity and characterized from Bacillus sp. strain MH-1. They had different molecular masses (71, 62, and 53 kDa), pIs (5.3, 4.8, and 4.7), and N-terminal amino acid sequences. Chitinases L, M, and S showed relatively high temperature optima (75, 65, and 75 degreesC) and stabilities and showed pH optima in an acidic range (pH 6.5, 5.5, and 5.5, respectively). When reacted with acetylchitohexaose [(GlcNAc)6], chitinases L and S produced (GlcNAc)2 at the highest rate while chitinase M produced (GlcNAc)3 at the highest rate. None of the three chitinases hydrolyzed (GlcNAc)2. Chitinase L produced (GlcNAc)2 and (GlcNAc)3 in most abundance from 66 and 11% partially acetylated chitosan. The p-nitrophenol (pNP)-releasing activity of chitinase L was highest toward pNP-(GlcNAc)2, and those of chitinases M and S were highest toward pNP-(GlcNAc)3. All three enzymes were inert to pNP-GlcNAc. AgCl, HgCl2, and (GlcNAc)2 inhibited the activities of all three enzymes, while MnCl2 and CaCl2 slightly activated all of the enzymes. PMID- 9726889 TI - Genetic relatedness among environmental, clinical, and diseased-eel Vibrio vulnificus isolates from different geographic regions by ribotyping and randomly amplified polymorphic DNA PCR. AB - Genetic relationships among 132 strains of Vibrio vulnificus (clinical, environmental, and diseased-eel isolates from different geographic origins, as well as seawater and shellfish isolates from the western Mediterranean coast, including reference strains) were analyzed by random amplified polymorphic DNA (RAPD) PCR. Results were validated by ribotyping. For ribotyping, DNAs were digested with KpnI and hybridized with an oligonucleotide probe complementary to a highly conserved sequence in the 23S rRNA gene. Random amplification of DNA was performed with M13 and T3 universal primers. The comparison between ribotyping and RAPD PCR revealed an overall agreement regarding the high level of homogeneity of diseased-eel isolates in contrast to the genetic heterogeneity of Mediterranean isolates. The latter suggests the existence of autochthonous clones present in Mediterranean coastal waters. Both techniques have revealed a genetic proximity among Spanish fish farm isolates and a close relationship between four Spanish eel farm isolates and some Mediterranean isolates. Whereas the differentiation within diseased-eel isolates was only possible by ribotyping, RAPD PCR was able to differentiate phenotypically atypical isolates of V. vulnificus. On the basis of our results, RAPD PCR is proposed as a better technique than ribotyping for rapid typing in the routine analysis of new V. vulnificus isolates. PMID- 9726890 TI - Genetic characterization and physiological role of endopeptidase O from Lactobacillus helveticus CNRZ32. AB - A previously identified insert expressing an endopeptidase from a Lactobacillus helveticus CNRZ32 genomic library was characterized. Nucleotide sequence analysis revealed an open reading frame of 1,941 bp encoding a putative protein of 71.2 kDa which contained a zinc-protease motif. Protein homology searches revealed that this enzyme has 40% similarity with endopeptidase O (PepO) from Lactococcus lactis P8-2-47. Northern hybridization revealed that pepO is monocistronic and is expressed throughout the growth phase. CNRZ32 derivatives lacking PepO activity were constructed via gene replacement. Enzyme assays revealed that the PepO mutant had significantly reduced endopeptidase activity when compared to CNRZ32 with two of the three substrates examined. Growth studies indicated that PepO has no detectable effect on growth rate or acid production by Lactobacillus helveticus CNRZ32 in amino acid defined or skim milk medium. PMID- 9726891 TI - Mode of action of linenscin OC2 against Listeria innocua. AB - Linenscin OC2 is a small hydrophobic substance produced by the orange cheese coryneform bacterium Brevibacterium linens OC2. Linenscin OC2 inhibits growth of gram-negative bacteria with an altered outer membrane permeability and gram positive bacteria. It is also able to lyse eucaryotic cells. The mode of action of linenscin OC2 on the Listeria innocua cytoplasmic membrane and the effects of environmental parameters were investigated. Addition of low doses of linenscin OC2 resulted in an immediate perturbation of the permeability properties of the cytoplasmic membrane and of the bacterial energetic state. Linenscin OC2 induced a loss of cytoplasmic potassium, depolarization of the cytoplasmic membrane, complete hydrolysis of internal ATP, efflux of inorganic phosphate, and transient increase in oxygen consumption. Potassium loss occurred in the absence of a proton motive force and was severely reduced at low temperatures, presumably as a result of increased ordering of the lipid hydrocarbon chains of the cytoplasmic membrane. We propose that linenscin OC2 interacts with the cytoplasmic membrane and that the permeability changes observed at low doses reflect the formation of pore-like structures in this membrane. PMID- 9726892 TI - Genotypic and phenotypic responses of a riverine microbial community to polycyclic aromatic hydrocarbon contamination. AB - The phenotypic and genotypic adaptation of a freshwater sedimentary microbial community to elevated (22 to 217 microgram [dry weight] of sediment-1) levels of polycyclic aromatic hydrocarbons (PAHs) was determined by using an integrated biomolecular approach. Central to the approach was the use of phospholipid fatty acid (PLFA) profiles to characterize the microbial community structure and nucleic acid analysis to quantify the frequency of degradative genes. The study site was the Little Scioto River, a highly impacted, channelized riverine system located in central Ohio. This study site is a unique lotic system, with all sampling stations having similar flow and sediment characteristics both upstream and downstream from the source of contamination. These characteristics allowed for the specific analysis of PAH impact on the microbial community. PAH concentrations in impacted sediments ranged from 22 to 217 microgram (dry weight) of sediment-1, while PAH concentrations in ambient sediments ranged from below detection levels to 1.5 microgram (dry weight) of sediment-1. Total microbial biomass measured by phospholipid phosphate (PLP) analysis ranged from 95 to 345 nmol of PLP g (dry weight) of sediment-1. Nucleic acid analysis showed the presence of PAH-degradative genes at all sites, although observed frequencies were typically higher at contaminated sites. Principal component analysis of PLFA profiles indicated that moderate to high PAH concentrations altered microbial community structure and that seasonal changes were comparable in magnitude to the effects of PAH pollution. These data indicate that this community responded to PAH contamination at both the phenotypic and the genotypic level. PMID- 9726893 TI - Assessment of reductive acetogenesis with indigenous ruminal bacterium populations and Acetitomaculum ruminis. AB - The objective of this study was to evaluate the role of reductive acetogenesis as an alternative H2 disposal mechanism in the rumen. H2/CO2-supported acetogenic ruminal bacteria were enumerated by using a selective inhibitor of methanogenesis, 2-bromoethanesulfonic acid (BES). Acetogenic bacteria ranged in density from 2.5 x 10(5) cells/ml in beef cows fed a high-forage diet to 75 cells/ml in finishing steers fed a high-grain diet. Negligible endogenous acetogenic activity was demonstrated in incubations containing ruminal contents, NaH13CO3, and 100% H2 gas phase since [U-13C]acetate, as measured by mass spectroscopy, did not accumulate. Enhancement of acetogenesis was observed in these incubations when methanogenesis was inhibited by BES and/or by the addition of an axenic culture of the rumen acetogen Acetitomaculum ruminis 190A4 (10(7) CFU/ml). To assess the relative importance of population density and/or H2 concentration for reductive acetogenesis in ruminal contents, incubations as described above were performed under a 100% N2 gas phase. Both selective inhibition of methanogenesis and A. ruminis 190A4 fortification (>10(5) CFU/ml) were necessary for the detection of reductive acetogenesis under H2-limiting conditions. Under these conditions, H2 accumulated to 4, 800 ppm. In contrast, H2 accumulated to 400 ppm in incubations with active methanogenesis (without BES). These H2 concentrations correlated well with the pure culture H2 threshold concentrations determined for A. ruminis 190A4 (3,830 ppm) and the ruminal methanogen 10-16B (126 ppm). The data demonstrate that ruminal methanogenic bacteria limited reductive acetogenesis by lowering the H2 partial pressure below the level necessary for H2 utilization by A. ruminis 190A4. PMID- 9726894 TI - Genetic analysis of Comamonas acidovorans polyhydroxyalkanoate synthase and factors affecting the incorporation of 4-hydroxybutyrate monomer. AB - The polyhydroxyalkanoate (PHA) synthase gene of Comamonas acidovorans DS-17 (phaCCa) was cloned by using the synthase gene of Alcaligenes eutrophus as a heterologous hybridization probe. Complete sequencing of a 4.0-kbp SmaI-HindIII (SH40) subfragment revealed the presence of a 1,893-bp PHA synthase coding region which was followed by a 1,182-bp beta-ketothiolase gene (phaACa). Both the translated products of these genes showed significant identity, 51.1 and 74.2%, respectively, to the primary structures of the products of the corresponding genes in A. eutrophus. The arrangement of PHA biosynthesis genes in C. acidovorans was also similar to that in A. eutrophus except that the third gene, phaB, coding for acetoacetyl-coenzyme A reductase, was not found in the region downstream of phaACa. The cloned fragment complemented a PHA-negative mutant of A. eutrophus, PHB-4, resulting in poly-3-hydroxybutyrate accumulation of up to 73% of the dry cell weight when fructose was the carbon source. The heterologous expression enabled the incorporation of 4-hydroxybutyrate (4HB) and 3 hydroxyvalerate monomers. The PHA synthase of C. acidovorans does not appear to show any preference for 4-hydroxybutyryl-coenzyme A as a substrate. This leads to the suggestion that in C. acidovorans, it is the metabolic pathway, and not the specificity of the organism's PHA synthase, that drives the incorporation of 4HB monomers, resulting in the efficient accumulation of PHA with a high 4HB content. PMID- 9726895 TI - New nitrogen-fixing microorganisms detected in oligotrophic oceans by amplification of Nitrogenase (nifH) genes. AB - Oligotrophic oceanic waters of the central ocean gyres typically have extremely low dissolved fixed inorganic nitrogen concentrations, but few nitrogen-fixing microorganisms from the oceanic environment have been cultivated. Nitrogenase gene (nifH) sequences amplified directly from oceanic waters showed that the open ocean contains more diverse diazotrophic microbial populations and more diverse habitats for nitrogen fixers than previously observed by classical microbiological techniques. Nitrogenase genes derived from unicellular and filamentous cyanobacteria, as well as from the alpha and gamma subdivisions of the class Proteobacteria, were found in both the Atlantic and Pacific oceans. nifH sequences that cluster phylogenetically with sequences from sulfate reducers or clostridia were found associated with planktonic crustaceans. Nitrogenase sequence types obtained from invertebrates represented phylotypes distinct from the phylotypes detected in the picoplankton size fraction. The results indicate that there are in the oceanic environment several distinct potentially nitrogen fixing microbial assemblages that include representatives of diverse phylotypes. PMID- 9726896 TI - Effect of nitrogen source on growth and trichloroethylene degradation by methane oxidizing bacteria. AB - The effect of nitrogen source on methane-oxidizing bacteria with respect to cellular growth and trichloroethylene (TCE) degradation ability were examined. One mixed chemostat culture and two pure type II methane-oxidizing strains, Methylosinus trichosporium OB3b and strain CAC-2, which was isolated from the chemostat culture, were used in this study. All cultures were able to grow with each of three different nitrogen sources: ammonia, nitrate, and molecular nitrogen. Both M. trichosporium OB3b and strain CAC-2 showed slightly lower net cellular growth rates and cell yields but exhibited higher methane uptake rates, levels of poly-beta-hydroxybutyrate (PHB) production, and naphthalene oxidation rates when grown under nitrogen-fixing conditions. The TCE-degrading ability of each culture was measured in terms of initial TCE oxidation rates and TCE transformation capacities (mass of TCE degraded/biomass inactivated), measured both with and without external energy sources. Higher initial TCE oxidation rates and TCE transformation capacities were observed in nitrogen-fixing mixed, M. trichosporium OB3b, and CAC-2 cultures than in nitrate- or ammonia-supplied cells. TCE transformation capacities were found to correlate with cellular PHB content in all three cultures. The results of this study suggest that the nitrogen-fixing capabilities of methane-oxidizing bacteria can be used to select for high-activity TCE degraders for the enhancement of bioremediation in fixed nitrogen-limited environments. PMID- 9726897 TI - Induction of acid resistance of Salmonella typhimurium by exposure to short-chain fatty acids. AB - Exposure to short-chain fatty acids (SCFA) is one of the stress conditions Salmonella typhimurium encounters during its life cycle, because SCFA have been widely used as food preservatives and SCFA are also present at high concentrations in the gastrointestinal tracts of host animals. The effects of SCFA on the acid resistance of the organism were examined in an attempt to understand the potential role of SCFA in the pathogenesis of S. typhimurium. The percent survival of S. typhimurium at pH 3.0 was determined after exposure to SCFA for 1 h at pH 7.0. The percent acid survival, which varied depending on the SCFA species and the concentration used, was 42 after exposure to 100 mM propionate at pH 7.0 under aerobic incubation conditions, while less than 1% could survive without exposure. The SCFA-induced acid resistance was markedly enhanced by anaerobiosis (64%), lowering pH conditions (138% at pH 5.0), or increasing incubation time (165% with 4 h) during exposure to propionic acid. When protein synthesis during exposure to propionate was blocked by chloramphenicol, the percent acid survival was less than 1, indicating that the protein synthesis induced by exposure to propionate is required for the induction of the acid resistance. The percent acid survival determined with the isogenic mutant strains defective in acid tolerance response revealed that AtrB protein is necessary for the full induction of acid resistance by exposure to propionate, while unexpectedly, inactivation of PhoP significantly increased acid resistance over that of the wild type (P < 0.05). The results suggest that the virulence of S. typhimurium may be enhanced by increasing acid resistance upon exposure to SCFA during its life cycle and further enhanced by anaerobiosis, low pH, and prolonged exposure time. PMID- 9726899 TI - Cellular events involved in survival of individual arbuscular mycorrhizal symbionts growing in the absence of the host AB - A survival strategy operating in the absence of the host was shown in obligately biotrophic arbuscular mycorrhizal (AM) symbionts. When no host-derived signals from the surrounding environment were perceived by germinating spores, fungal hyphae underwent a programmed growth arrest and resource reallocation, allowing long-term maintenance of viability and host infection capability. The early stages of mycelial growth of AM fungi were studied by a combination of time-lapse and video-enhanced light microscopy, image analysis, and immunodetection, with the aim of acquiring knowledge of cell events leading to the arrest of mycelial growth. The time-course of growth arrest was resolved by precisely timing the growth rate and magnitude of the mycelium originating from individual spores of Glomus caledonium. Extensive mycelial growth was observed during the first 15 days; thereafter, fungal hyphae showed retraction of protoplasm from the tips, with formation of retraction septa separating viable from empty hyphal segments. This active process involved migration of nuclei and cellular organelles and appeared to be functional in the ability of the fungus to survive in the absence of a host. Immunodetection of cytoskeletal proteins, metabolic activity, and the retention of infectivity of germinated spores confirmed the developmental data. The highest amounts of tubulins were detected when hyphal growth had ceased but when retraction of protoplasm was most active. This was consistent with the role of the cytoskeleton during protoplasm retraction. Succinate dehydrogenase activity in hyphae proximal to the mother spore was still detectable in 6-month old mycelium, which remained viable and able to form appressoria and produce symbiotic structures. PMID- 9726898 TI - Genetic diversity of the biofilm covering Montacuta ferruginosa (Mollusca, bivalvia) as evaluated by denaturing gradient gel electrophoresis analysis and cloning of PCR-amplified gene fragments coding for 16S rRNA. AB - The shell of the bivalve Montacuta ferruginosa, a symbiont living in the burrow of an echinoid, is covered with a rust-colored biofilm. This biofilm includes different morphotypes of bacteria that are encrusted with a mineral rich in ferric ion and phosphate. The aim of this research was to determine the genetic diversity and phylogenetic affiliation of the biofilm bacteria. Also, the possible roles of the microorganisms in the processes of mineral deposition within the biofilm, as well as their impact on the biology of the bivalve, were assessed by phenotypic inference. The genetic diversity was determined by denaturing gradient gel electrophoresis (DGGE) analysis of short (193-bp) 16S ribosomal DNA PCR products obtained with primers specific for the domain Bacteria. This analysis revealed a diverse consortium; 11 to 25 sequence types were detected depending on the method of DNA extraction used. Individual biofilms analyzed by using the same DNA extraction protocol did not produce identical DGGE profiles. However, different biofilms shared common bands, suggesting that similar bacteria can be found in different biofilms. The phylogenetic affiliations of the sequence types were determined by cloning and sequencing the 16S rRNA genes. Close relatives of the genera Pseudoalteromonas, Colwellia, and Oceanospirillum (members of the gamma-Proteobacteria lineage), as well as Flexibacter maritimus (a member of the Cytophaga-Flavobacter-Bacteroides lineage), were found in the biofilms. We inferred from the results that some of the biofilm bacteria could play a role in the mineral formation processes. PMID- 9726902 TI - Symbiotic relationship of thiothrix spp. with An echinoderm AB - Immunoassay procedures were used to investigate the symbiotic relationship of Thiothrix spp. in the intestinal cecum of the spatangoid species Echinocardium cordatum. Thiothrix spp. were identified in nodule samples from E. cordatum digestive tubes based on microscopic examination, enzyme-linked immunosorbent assay, and indirect immunofluorescence. Thiothrix spp. protein made up as much as 84% of the total protein content of the nodules. This is the first identification of Thiothrix spp. internally symbiotic with marine invertebrates. PMID- 9726901 TI - Biofilms on indwelling urethral catheters produce quorum-sensing signal molecules in situ and in vitro. AB - Acylated homoserine lactones (AHLs) are chemical signals that mediate population density-dependent (quorum-sensing) gene expression in numerous gram-negative bacteria. In this study, gram-negative bacilli isolated from catheters were screened for AHL production by a cross-feeding assay utilizing an AHL-responsive Agrobacterium tumefaciens reporter strain. Positive reactions were obtained from 14 isolates of Pseudomonas aeruginosa; negative or weakly positive reactions were recorded for isolates of five other species. P. aeruginosa biofilms were then produced on catheters in a physical model of the bladder. Sections of colonized all-silicone catheters gave positive reactions for the quorum-sensing signal molecules as did sections that had been cleaned of biofilm and autoclaved. Control sections of unused catheters were negative in the tests. Sections from four of nine catheters that had been freshly removed from patients gave positive reactions for AHLs. Cleaned autoclaved sections of three of these catheters also gave strongly positive reactions for AHLs. These results demonstrate that AHLs are produced by biofilms as they develop on the catheters both in vitro in the model and in vivo in the patient's bladder. They represent the first demonstration of AHL production by biofilms in a clinical setting. PMID- 9726900 TI - Identification and activities in situ of Nitrosospira and Nitrospira spp. as dominant populations in a nitrifying fluidized bed reactor. AB - Bacterial aggregates from a chemolithoautotrophic, nitrifying fluidized bed reactor were investigated with microsensors and rRNA-based molecular techniques. The microprofiles of O2, NH4+, NO2-, and NO3- demonstrated the occurrence of complete nitrification in the outer 125 microgram of the aggregates. The ammonia oxidizers were identified as members of the Nitrosospira group by fluorescence in situ hybridization (FISH). No ammonia- or nitrite-oxidizing bacteria of the genus Nitrosomonas or Nitrobacter, respectively, could be detected by FISH. To identify the nitrite oxidizers, a 16S ribosomal DNA clone library was constructed and screened by denaturing gradient gel electrophoresis and selected clones were sequenced. The organisms represented by these sequences formed two phylogenetically distinct clusters affiliated with the nitrite oxidizer Nitrospira moscoviensis. 16S rRNA-targeted oligonucleotide probes were designed for in situ detection of these organisms. FISH analysis showed that the dominant populations of Nitrospira spp. and Nitrosospira spp. formed separate, dense clusters which were in contact with each other and occurred throughout the aggregate. A second, smaller, morphologically and genetically different population of Nitrospira spp. was restricted to the outer nitrifying zones. PMID- 9726903 TI - Genetically modified ruminal bacteria protect sheep from fluoroacetate poisoning. AB - Four strains of Butyrivibrio fibrisolvens, transformed with a gene encoding fluoroacetate dehalogenase, maintained a combined population of 10(6) to 10(7) cells ml-1 in the rumens of test sheep. Five inoculated sheep showed markedly reduced toxicological symptoms after fluoroacetate poisoning when behavioral, physiological, and histological effects were compared with those of five uninoculated control sheep. PMID- 9726904 TI - Antibiotic resistance in Acinetobacter spp. isolated from sewers receiving waste effluent from a hospital and a pharmaceutical plant. AB - The possible increase of antibiotic-resistant bacteria in sewage associated with the discharge of wastewater from a hospital and a pharmaceutical plant was investigated by using Acinetobacter species as environmental bacterial indicators. The level of susceptibility to six antimicrobial agents was determined in 385 Acinetobacter strains isolated from samples collected upstream and downstream from the discharge points of the hospital and the pharmaceutical plant. Results indicated that while the hospital waste effluent affected only the prevalence of oxytetracycline resistance, the discharge of wastewater from the pharmaceutical plant was associated with an increase in the prevalence of both single- and multiple-antibiotic resistance among Acinetobacter species in the sewers. PMID- 9726905 TI - Influence of growth mode and sucrose on susceptibility of Streptococcus sanguis to amine fluorides and amine fluoride-inorganic fluoride combinations. AB - This study evaluated the susceptibility to amine fluorides (AmFs) of planktonic and biofilm cultures of Streptococcus sanguis grown with and without sucrose. Cultures were incubated with AmFs (250 mg of fluoride liter-1) for 1 min. The susceptibility of biofilms was less than that of the planktonic form and was further decreased by growth in the presence of sucrose. PMID- 9726907 TI - Chemostat production of plantaricin C by Lactobacillus plantarum LL441. AB - Plantaricin C, a bacteriocin synthesized by Lactobacillus plantarum LL441, was optimally produced in chemostats kept at pH 5.0, 30 degreesC, 150 rpm, and a dilution rate of 0.05 h-1 when glucose was used as carbon source and a dilution rate of 0.10 to 0.12 h-1 when sucrose or fructose was used instead. Production was abolished at high dilution rates, i.e., when the cells grew rapidly in all carbon sources. PMID- 9726906 TI - Pseudomonas sp. strain 273, an aerobic alpha, omega-dichloroalkaneDegrading bacterium. AB - A gram-negative, aerobic bacterium was isolated from soil; this bacterium grew in 50% (vol/vol) suspensions of 1,10-dichlorodecane (1,10-DCD) as the sole source of carbon and energy. Phenotypic and small-subunit ribosomal RNA characterizations identified the organism, designated strain 273, as a member of the genus Pseudomonas. After induction with 1,10-DCD, Pseudomonas sp. strain 273 released stoichiometric amounts of chloride from C5 to C12 alpha, omega-dichloroalkanes in the presence of oxygen. No dehalogenation occurred under anaerobic conditions. The best substrates for dehalogenation and growth were C9 to C12 chloroalkanes. The isolate also grew with nonhalogenated aliphatic compounds, and decane-grown cells dechlorinated 1,10-DCD without a lag phase. In addition, cells grown on decane dechlorinated 1,10-DCD in the presence of chloramphenicol, indicating that the 1,10-DCD-dechlorinating enzyme system was also induced by decane. Other known alkane-degrading Pseudomonas species did not grow with 1,10-DCD as a carbon source. Dechlorination of 1,10-DCD was demonstrated in cell extracts of Pseudomonas sp. strain 273. Cell-free activity was strictly oxygen dependent, and NADH stimulated dechlorination, whereas EDTA had an inhibitory effect. PMID- 9726908 TI - Composition and susceptibility to chlorhexidine of multispecies biofilms of oral bacteria. AB - Using a constant-depth film fermentor, we have grown a six-membered biofilm community with a bacterial composition similar to that found in supragingival dental plaque. Cryosectioning revealed the distribution of bacteria throughout the biofilm. Exposure to 0.2% chlorhexidine for up to 5 min had little effect on biofilm viability. PMID- 9726909 TI - Prevalence of the Rhizobium etli-like allele in genes coding for 16S rRNA among the indigenous rhizobial populations found associated with wild beans from the Southern Andes in Argentina. AB - A collection of rhizobial isolates from nodules of wild beans, Phaseolus vulgaris var. aborigineus, found growing in virgin lands in 17 geographically separate sites in northwest Argentina was characterized on the basis of host range, growth, hybridization to a nifH probe, analysis of genes coding for 16S rRNA (16S rDNA), DNA fingerprinting, and plasmid profiles. Nodules in field-collected wild bean plants were largely dominated by rhizobia carrying the 16S rDNA allele of Rhizobium etli. A similar prevalence of the R. etli allele was observed among rhizobia trapped from nearby soil. Intragroup diversity of wild bean isolates with either R. etli-like or Rhizobium leguminosarum bv. phaseoli-like alleles was generally found across northwest Argentina. The predominance of the R. etli allele suggests that in this center of origin of P. vulgaris the coevolution of Rhizobium spp. and primitive beans has resulted in this preferential symbiotic association. PMID- 9726910 TI - Discrimination of psychrotrophic and mesophilic strains of the Bacillus cereus group by PCR targeting of major cold shock protein genes. AB - Detection of psychrotrophic strains (those able to grow at or below 7 degreesC) of the Bacillus cereus group (Bacillus cereus, Bacillus thuringiensis, and Bacillus mycoides) in food products is at present extremely slow with conventional microbiology. This is due to an inability to discriminate these cold adapted strains from their mesophilic counterparts (those able to grow only above 7 degreesC) by means other than growth at low temperature, which takes 5 to 10 days for detection. Here we report the development of a single PCR assay that, using major cold shock protein-specific primers and appropriate annealing temperatures, is capable of both rapidly identifying bacteria of the B. cereus group and discriminating between psychrotrophic and mesophilic strains. It is intended that this development help to more accurately predict the shelf life of refrigerated pasteurized food and dairy products and to reduce the incidence of food poisoning by psychrotrophic strains of the B. cereus group. PMID- 9726912 TI - Purification and properties of two thermostable alkaline xylanases from an alkaliphilic bacillus sp AB - Two xylanases, designated XylA and XylB, were purified from the culture supernatant of the alkaliphilic Bacillus sp. strain AR-009. The molecular masses of the two enzymes were estimated to be 23 kDa (XylA) and 48 kDa (XylB) by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The optimum pHs for activity were 9 for XylA and 9 to 10 for XylB. The temperature optima for the activity of XylA were 60 degreesC at pH 9 and 70 degreesC at pH 8. XylB was optimally active at 75 degreesC at pH 9 and 70 degreesC at pH 8. Both enzymes were stable in a broad pH range and showed good stability when incubated at 60 and 65 degreesC in pH 8 and 9 buffers. PMID- 9726911 TI - Influence of lipid composition on pediocin PA-1 binding to phospholipid vesicles. AB - Pediocin PA-1 bound to anionic lipid vesicles with saturated or unsaturated fatty acid chains in a lipid concentration-dependent fashion. Little change in binding parameters was observed for zwitterionic lipid vesicles. Decreasing the anionic lipid content of the vesicles gave a higher relative dissociation constant for the peptide-lipid interactions and further supports the electrostatic interaction model of binding. PMID- 9726913 TI - Expression of phanerochaete chrysosporium genes encoding lignin peroxidases, manganese peroxidases, and glyoxal oxidase in wood AB - Expression of Phanerochaete chrysosporium genes encoding ligninolytic enzymes was assessed in wood. Poly(A) RNA was extracted from colonized wood chips by magnetic capture, and specific transcripts were quantified by competitive reverse transcriptase PCR. mRNA levels varied substantially among lignin peroxidase genes, and transcript patterns were dramatically different from those in previous studies with defined media. PMID- 9726914 TI - Diversity of frankia strains in root nodules of plants from the families elaeagnaceae and rhamnaceae AB - Partial 16S ribosomal DNAs (rDNAs) were PCR amplified and sequenced from Frankia strains living in root nodules of plants belonging to the families Elaeagnaceae and Rhamnaceae, including Colletia hystrix, Elaeagnus angustifolia, an unidentified Elaeagnus sp., Talguenea quinquenervia, and Trevoa trinervis. Nearly full-length 16S rDNAs were sequenced from strains of Frankia living in nodules of Ceanothus americanus, C. hystrix, Coriaria arborea, and Trevoa trinervis. Partial sequences also were obtained from Frankia strains isolated and cultured from the nodules of C. hystrix, Discaria serratifolia, D. trinervis, Retanilla ephedra, T. quinquenervia, and T. trinervis (Rhamnaceae). Comparison of these sequences and other published sequences of Frankia 16S rDNA reveals that the microsymbionts and isolated strains from the two plant families form a distinct phylogenetic clade, except for those from C. americanus. All sequences in the clade have a common 2 base deletion compared with other Frankia strains. Sequences from C. americanus nodules lack the deletion and cluster with Frankia strains infecting plants of the family Rosaceae. Published plant phylogenies (based on chloroplast rbcL sequences) group the members of the families Elaeagnaceae and Rhamnaceae together in the same clade. Thus, with the exception of C. americanus, actinorhizal plants of these families and their Frankia microsymbionts share a common symbiotic origin. PMID- 9726915 TI - Modeling nonlinear red cell elasticity. PMID- 9726916 TI - Run, don't hop, through the nearest calcium channel. PMID- 9726917 TI - Theoretical analysis of the Ca2+ spark amplitude distribution. AB - A difficulty of using confocal microscopy to study Ca2+ sparks is the uncertainty of the linescan position with respect to the source of Ca2+ release. Random placement of the linescan is expected to result in a broad distribution of measured Ca2+ spark amplitudes (a) even if all Ca2+ sparks were generated identically. Thus variations in Ca2+ spark amplitude due to positional differences between confocal linescans and Ca2+ release site are intertwined with variations due to intrinsic differences in Ca2+ release properties. To separate these two sources of variations on the Ca2+ spark amplitude, we determined the effect changes of channel current or channel open time--collectively called the source strength, alpha--had on the measured Ca2+ spark amplitude histogram, N(a). This was done by 1) simulating Ca2+ release, Ca2+ and fluo-3 diffusion, and Ca2+ binding reactions; 2) simulation of image formation of the Ca2+ spark by a confocal microscope; and 3) using a novel automatic Ca2+ spark detector. From these results we derived an integral equation relating the probability density function of source strengths, f alpha (alpha), to N(a), which takes into account random positional variations between the source and linescan. In the special, but important, case that the spatial distribution of Ca(2+)-bound fluo-3 is Gaussian, we show the following: 1) variations of Ca2+ spark amplitude due to positional or intrinsic differences can be separated, and 2) f alpha (alpha) can, in principle, be calculated from the Ca2+ spark amplitude histogram since N(a) is the sum of shifted hyperbolas, where the magnitudes of the shifts and weights depend on f alpha (alpha). In particular, if all Ca2+ sparks were generated identically, then the plot of 1/N(a) against a will be a straight line. Multiple populations of channels carrying distinct currents are revealed by discontinuities in the 1/N(a) plot. 3) Although the inverse relationship between Ca2+ spark amplitude and decay time might be used to distinguish Ca2+ sparks from different channel populations, noise can render the measured decay times meaningless for small amplitude Ca2+ sparks. PMID- 9726918 TI - Structural studies of a stable parallel-stranded DNA duplex incorporating isoguanine:cytosine and isocytosine:guanine basepairs by nuclear magnetic resonance spectroscopy. AB - Isoguanine (2-hydroxyladenine) is a product of oxidative damage to DNA and has been shown to cause mutation. It is also a potent inducer of parallel-stranded DNA duplex structure. The structure of the parallel-stranded DNA duplex (PS duplex) 5'-d(TiGiCAiCiGiGAiCT) + 5'-d(ACGTGCCTGA), containing the isoguanine (iG) and 5-methyl-isocytosine (iC) bases, has been determined by NMR refinement. All imino protons associated with the iG:C, G:iC, and A:T (except the two terminal A:T) basepairs are observed at 2 degrees C, consistent with the formation of a stable duplex suggested by the earlier Tm measurements [Sugiyama, H., S. Ikeda, and I. Saito. 1996. J. Am. Chem. Soc. 118:9994-9995]. All basepairs are in the reverse Watson-Crick configuration. The structural characteristics of the refined PS-duplex are different from those of B-DNA. The PS duplex has two grooves with similar width (7.0 A) and depth (7.7 A), in contrast to the two distinct grooves (major groove width 11.7 A, depth 8.5 A, and minor groove width 5.7 A, depth 7.5 A) of B-DNA. The resonances of the amino protons of iG and C are clearly resolved and observable, but those of the G and iC are very broad and difficult to observe. Several intercalators with different complexities, including ethidium, daunorubicin, and nogalamycin, have been used to probe the flexibility of the backbone of the (iG, iC)-containing PS-duplex. All of them produce drug-induced UV/vis spectra identical to their respective spectra when bound to B-DNA, suggesting that those drugs bind to the (iG, iC)-containing PS-duplex using similar intercalation processes. The results may be useful in the design of intercalator-conjugated oligonucleotides for antisense applications. The study presented in this paper augments our understanding of a growing number of parallel-stranded DNA structures, including the G-quartet, the i-motif, and the unusual homo basepaired parallel-stranded double helix. Their possible relevance is discussed. PMID- 9726919 TI - Structural and morphological characterization of ultralente insulin crystals by atomic force microscopy: evidence of hydrophobically driven assembly. AB - Although x-ray crystal structures exist for many forms of insulin, the hormone involved in glucose metabolism and used in the treatment of diabetes, x-ray structural characterization of therapeutically important long-acting crystalline ultralente insulin forms has been elusive because of small crystal size and poor diffraction characteristics. We describe tapping-mode atomic force microscopy (TMAFM) studies, performed directly in crystallization liquor, of ultralente crystals prepared from bovine, human, and porcine insulins. Lattice images obtained from direct imaging of crystal planes are consistent with R3 space group symmetry for each insulin type, but the morphology of the human and porcine crystals observed by AFM differs substantially from that of the bovine insulin crystals. Human and porcine ultralente crystals exhibited large, molecularly flat (001) faces consisting of hexagonal arrays of close packed hexamers. In contrast, bovine ultralente crystals predominantly exhibited faces with cylindrical features assignable to close-packed stacks of insulin hexamers laying in-plane, consistent with the packing motif of the (010) and (011) planes. This behavior is attributed to a twofold increase in the hydrophobic character of the upper and lower surfaces of the donut-shaped insulin hexamer in bovine insulin compared to its human and porcine counterparts that results from minor sequence differences between these insulins. The increased hydrophobicity of these surfaces can promote hexamer-hexamer stacking in precrystalline aggregates or enhance attachment of single hexamers along the c axis at the crystal surface during crystal growth. Both events lead to enhanced growth of ?hk0? planes instead of (001). The insulin hexamers on the (010) and (110) faces are exposed "edge-on" to the aqueous medium, such that solvent access to the center of the hexamer and to solvent channels is reduced compared to the (001) surface, consistent with the slower dissolution and reputed unique basal activity of bovine ultralente insulin. These observations demonstrate that subtle variations in amino acid sequence can dramatically affect the interfacial structure of crystalline proteins. PMID- 9726921 TI - Autocorrelation function and power spectrum of two-state random processes used in neurite guidance. AB - During development neurons extend and retract cytoskeletal structures, chiefly microtubules and filopodia, to process informational cues from the extracellular environment and thereby guide growth cone migration toward an appropriate synaptic partner. This cytoskeleton-based exploration is achieved by stochastic switching, with microtubules and filopodia alternating between growing and shortening phases apparently at random. If stabilizing signals are not detected during the growth phase, then the structures switch to a shortening state, from which they can again return to a growth phase, and so forth. A useful means of characterizing these stochastic processes in a model-independent way is by autocorrelation and spectral analysis. Previously, we compared experiment to theory by performing Monte Carlo simulations and computing the autocorrelation function and power spectrum from the simulated dynamics, an approach that is computationally intensive and requires recalculation whenever model parameters are changed. Here we present analytical expressions for the autocorrelation function and power spectrum, which compactly characterize microtubule and filopodial dynamics based on the stochastic, two-state model. The model assumes that the phase times are of variable duration and gamma-distributed, consistent with experimental evidence for microtubules assembled in vitro from purified tubulin. The analytical expressions permit the precise quantitative characterization of changes in microtubule and filopodial searching behavior corresponding to changes in the shape of the gamma distribution. PMID- 9726920 TI - General model for lipid-mediated two-dimensional array formation of membrane proteins: application to bacteriorhodopsin. AB - Based on experimental evidence for 2D array formation of bacteriorhodopsin, we propose a general model for lipid-mediated 2D array formation of membrane proteins in lipid bilayers. The model includes two different lipid species, "annular" lipids and "neutral" lipids, and one protein species. The central assumption of the model is that the annular lipids interact more strongly with the protein than with the neutral lipids. Monte Carlo simulations performed on this model show that 2D arrays of proteins only form when there are annular lipids present. In addition, no arrays form if all of the lipids present are annular lipids. The geometry of the observed arrays is for the most part hexagonal. However, for a certain range of low annular lipid/protein ratios, arrays form that have geometries other than hexagonal. Using the assumption that the hydrocarbon chains of the annular lipids are restricted in motion when close to a protein, we expand the model to include a ground state and an excited state of the annular lipids. The main result from the extended model is that within a certain temperature range, increasing the temperature will lead to larger and more regular protein arrays. PMID- 9726922 TI - Theory for the hydrodynamic and electrophoretic stretch of tethered B-DNA. AB - We have developed a theory for the extension and force of B-DNA tethered at a fixed point in a uniform hydrodynamic flow or in a uniform applied electric field. The chain tethered in an electric field is considered to be subject to free electrophoresis compensated by free sedimentation in the opposite direction. This allows the use of results of free electrophoresis for including the effects of small ions. The force on the chain is derived for a sequence of ellipsoidal segments, each twice the persistence length of the wormlike chain. Hydrodynamic interaction between these segments is based on the long-range limit of flow around the prolate ellipsoids, as derived from equivalent Stokes spheres. The chain extension is derived by applying the entropic elasticity relation of Marko and Siggia (1995 Macromolecules. 28:8759-8770) to each segment for polymer chains under constant tension. We justify this procedure by comparing with extension results based on the Boltzmann averaged orientation of straight, freely jointed segments. Predicted results agree well with recent extension-flow experiments by Perkins et al., 1995. Science. 258:83-87, and with electrophoretic stretch experiments by Smith and Bendich (1990 Biopolymers. 29:1167-1173) on fluorescently stained B-DNA. We find that the equivalence of hydrodynamic and electrophoretic stretch, proposed by Long et al. (1996 Phys. Rev. Lett. 76:3858 3861; 1996 Biopolymers 39:755-759), is valid only for very small chain deformations, but not in general. PMID- 9726923 TI - Electrostatics and the ion selectivity of ligand-gated channels. AB - The nicotinic acetylcholine receptor (nAChR) is a cation-selective ion channel that opens in response to acetylcholine binding. The related glycine receptor (GlyR) is anion selective. The pore-lining domain of each protein may be modeled as a bundle of five parallel M2 helices. Models of the pore-lining domains of homopentameric nAChR and GlyR have been used in continuum electrostatics calculations to probe the origins of ion selectivity. Calculated pKA values suggest that "rings" of acidic or basic side chains at the mouths of the nAChR or GlyR M2 helix bundles, respectively, may not be fully ionized. In particular, for the nAChR the ring of glutamate side chains at the extracellular mouth of the pore is predicted to be largely protonated at neutral pH, whereas those glutamate side chains in the intracellular and intermediate rings (at the opposite mouth of the pore) are predicted to be fully ionized. Inclusion of the other domains of each protein represented as an irregular cylindrical tube in which the M2 bundles are embedded suggests that both the M2 helices and the extramembrane domains play significant roles in determining ion selectivity. PMID- 9726924 TI - Hexagonally packed DNA within bacteriophage T7 stabilized by curvature stress. AB - A continuum computation is proposed for the bending stress stabilizing DNA that is hexagonally packed within bacteriophage T7. Because the inner radius of the DNA spool is rather small, the stress of the curved DNA genome is strong enough to balance its electrostatic self-repulsion so as to form a stable hexagonal phase. The theory is in accord with the microscopically determined structure of bacteriophage T7 filled with DNA within the experimental margin of error. PMID- 9726925 TI - Trapping electrophoresis and ratchets: a theoretical study for DNA-protein complexes. AB - Recently, Griess and Serwer (1998. Biophys. J. 74:A71) showed that it was possible to use trapping electrophoresis and unbiased but asymmetrical electric field pulses to build a correlation ratchet that would allow the efficient separation of naked DNAs from identical DNAs that form a complex with a bulky object such as a protein. Here we present a theoretical investigation of this novel macromolecular separation process. We start by looking at the general features of this electrophoretic ratchet mechanism in the zero-frequency limit. We then examine the effects of finite frequencies on velocity and diffusion. Finally, we use the biased reptation model and computer simulations to understand the band-broadening processes. Our study establishes the main experimental regimes that can provide good resolution for specific applications. PMID- 9726926 TI - Evaluation of the electrostatic field strength at the site of exocytosis in adrenal chromaffin cells. AB - Exocytosis in secretory cells consists of release from intracellular storage granules directly into the extracellular space via fusion of the granule membrane with the plasma membrane of the cell. It is considered here as comprising two distinct processes. One is the close apposition of granule and plasma membranes. The other arises from interactions between the two membranes during the process of apposition, leading to the formation of a fusion pore. In the following it is shown for the case of the adrenal medullary chromaffin cell that the fusion pore can be ascribed to electroporation of the granule membrane, triggered by the strong electric field existing at the site of exocytosis. Based on an electric surface charge model of the cytoplasmic side of the plasma membrane, resulting from the negatively charged phosphatidylserine groups, it is found that the electrostatic field strength at the site of exocytosis reaches values on the order of 10(8) V/m at small intermembrane distances of 3 nm and lower. The field strength increases with the size of the disc-shaped plasma membrane region generating the electric field, reaching an approximate limit for a radius of 10 nm, at a surface charge density of 5.4 x 10(-2) C/m2. According to previous experimental evaluations of threshold field strength, this field is sufficiently strong to cause membrane electroporation. This step is a precondition for the subsequent membrane fusion during the ongoing process of apposition, leading to secretion. PMID- 9726928 TI - Nonlinear and asymmetric open channel characteristics of an ion-selective porin in planar membranes. AB - The open channel characteristics of the bacterial porin Omp32 from Comamonas acidovorans were investigated by means of conductance measurements in planar lipid bilayers of the Montal-Mueller type. Particularly at low salt conditions (< or = 30 mM KCl) Omp32 exhibited some unusual asymmetric and nonlinear functional properties. Current-voltage relationship measurements showed that conductance depends on the orientation of porin molecules and is a nonlinear function of the applied membrane potential. Conductance also depends on the salt concentration in a manner not common to porins and the salt concentration modulates the nonlinearity of conductance-voltage relationships. Omp32 is strongly anion selective. The nonlinear and asymmetric conductance of the open channel is a new observation in porins. PMID- 9726927 TI - Mechanism and specificity of lanthanide series cation transport by ionophores A23187, 4-BrA23187, and ionomycin. AB - A23187, 4-BrA23187, and ionomycin transport several lanthanide series trivalent cations at efficiencies similar to Ca2+, when compared at cation concentrations of approximately 10(-5) M, ionophore concentrations of approximately 10(-6) M, and a pH of 7.00. Selectivity sequences and the range of relative rates are as follows: A23187, Nd3+ > La3+ > Eu3+ > Gd3+ > Er3+ > Yb3+ > Lu3+ (approximately 34 fold); 4-BrA23187, Nd3+ > Eu3+ > Gd3+ > La3+ > Er3+ > Yb3+ > Lu3+ (approximately 34-fold); ionomycin, La3+ > Yb3+ > Nd3+ > Lu3+ > Er3+ > Eu3+ > Gd3+ (approximately 4-fold). At concentrations between 9 and 250 microM, La3+ is transported by an electroneutral mechanism, predominately through mixed complexes of the type (ionophore)2La-OH (A23187 and 4-BrA23187) or (ionophore)La-OH (ionomycin), when no membrane potential is present. For all three ionophores, an induced potential of approximately 160 mV accelerates transport by approximately 50-100%. However, measured values of H+/La3+ exchange indicate that only 4 BrA23187 displays a significant electrogenic activity under these conditions. At a La3+ concentration of 17 mM, transport by all three ionophores is electroneutral and apparently occurs through complexes of type (ionophore)3La (A23187 and 4-BrA23187) or (ionophore)La-OH (ionomycin). Analysis of these patterns in a context of comproportionation equilibria involving the transporting species and free La3+ indicates that the species containing three ionophore molecules are formed on the membrane when aqueous phase solution conditions would strongly favor a 1:1 complex, based upon previous studies in solution. The implications of this and other findings are discussed. PMID- 9726929 TI - Shaker and ether-a-go-go K+ channel subunits fail to coassemble in Xenopus oocytes. AB - Members of different voltage-gated K+ channel subfamilies usually do not form heteromultimers. However, coassembly between Shaker and ether-a-go-go (eag) subunits, members of two distinct K+ channel subfamilies, was suggested by genetic and functional studies (Zhong and Wu. 1991. Science. 252: 1562-1564; Chen, M.-L., T. Hoshi, and C.-F. Wu. 1996. Neuron. 17:535-542). We investigated whether Shaker and eag form heteromultimers in Xenopus laevis oocytes using electrophysiological and biochemical approaches. Coexpression of Shaker and eag subunits produced K+ currents that were virtually identical to the sum of separate Shaker and eag currents, with no change in the kinetics of Shaker inactivation. According to the results of dominant negative and reciprocal coimmunoprecipitation experiments, the Shaker and eag proteins do not interact. We conclude that Shaker and eag do not coassemble to form heteromultimers in Xenopus oocytes. PMID- 9726930 TI - Multiple channels mediate calcium leakage in the A7r5 smooth muscle-derived cell line. AB - Ca2+ entry under resting conditions may be important for contraction of vascular smooth muscle, but little is known about the mechanisms involved. Ca2+ leakage was studied in the A7r5 smooth muscle-derived cell line by patch-clamp techniques. Two channels that could mediate calcium influx at resting membrane potentials were characterized. In 110 mM Ba2+, one channel had a slope conductance of 6.0 +/- 0.6 pS and an extrapolated reversal potential of +41 +/- 13 mV (mean +/- SD, n = 8). The current rectified strongly, with no detectable outward current, even at +90 mV. Channel gating was voltage independent. A second type of channel had a linear current-voltage relationship, a slope conductance of 17.0 +/- 3.2 pS, and a reversal potential of +7 +/- 4 mV (n = 9). The open probability increased e-fold per 44 +/- 10 mV depolarization (n = 5). Both channels were also observed in 110 mM Ca2+. Noise analysis of whole-cell currents indicates that approximately 100 6-pS channels and 30 17-pS channels are open per cell. These 6-pS and 17-pS channels may contribute to resting calcium entry in vascular smooth muscle cells. PMID- 9726931 TI - Ion permeation and glutamate residues linked by Poisson-Nernst-Planck theory in L type calcium channels. AB - L-type Ca channels contain a cluster of four charged glutamate residues (EEEE locus), which seem essential for high Ca specificity. To understand how this highly charged structure might produce the currents and selectivity observed in this channel, a theory is needed that relates charge to current. We use an extended Poisson-Nernst-Planck (PNP2) theory to compute (mean) Coulombic interactions and thus to examine the role of the mean field electrostatic interactions in producing current and selectivity. The pore was modeled as a central cylinder with tapered atria; the cylinder (i.e., "pore proper") contained a uniform volume density of fixed charge equivalent to that of one to four carboxyl groups. The pore proper was assigned ion-specific, but spatially uniform, diffusion coefficients and excess chemical potentials. Thus electrostatic selection by valency was computed self-consistently, and selection by other features was also allowed. The five external parameters needed for a system of four ionic species (Na, Ca, Cl, and H) were determined analytically from published measurements of thre limiting conductances and two critical ion concentrations, while treating the pore as a macroscopic ion-exchange system in equilibrium with a uniform bath solution. The extended PNP equations were solved with these parameters, and the predictions were compared to currents measured in a variety of solutions over a range of transmembrane voltages. The extended PNP theory accurately predicted current-voltage relations, anomalous mole fraction effects in the observed current, saturation effects of varied Ca and Na concentrations, and block by protons. Pore geometry, dielectric permittivity, and the number of carboxyl groups had only weak effects. The successful prediction of Ca fluxes in this paper demonstrates that ad hoc electrostatic parameters, multiple discrete binding sites, and logistic assumptions of single-file movement are all unnecessary for the prediction of permeation in Ca channels over a wide range of conditions. Further work is needed, however, to understand the atomic origin of the fixed charge, excess chemical potentials, and diffusion coefficients of the channel. The Appendix uses PNP2 theory to predict ionic currents for published "barrier-and-well" energy profiles of this channel. PMID- 9726933 TI - Two-dimensional crystallization of Ca-ATPase by detergent removal. AB - By using Bio-Beads as a detergent-removing agent, it has been possible to produce detergent-depleted two-dimensional crystals of purified Ca-ATPase. The crystallinity and morphology of these different crystals were analyzed by electron microscopy under different experimental conditions. A lipid-to-protein ratio below 0.4 w/w was required for crystal formation. The rate of detergent removal critically affected crystal morphology, and large multilamellar crystalline sheets or wide unilamellar tubes were generated upon slow or fast detergent removal, respectively. Electron crystallographic analysis indicated unit cell parameters of a = 159 A, b = 54 A, and gamma = 90 degrees for both types of crystals, and projection maps at 15-A resolution were consistent with Ca ATPase molecules alternately facing the two sides of the membrane. Crystal formation was also affected by the protein conformation. Indeed, tubular and multilamellar crystals both required the presence of Ca2+; the presence of ADP gave rise to another type of packing within the unit cell (a = 86 A, b = 77 A, and gamma = 90 degrees), while maintaining a bipolar orientation of the molecules within the bilayer. All of the results are discussed in terms of nucleation and crystal growth, and a model of crystallogenesis is proposed that may be generally true for asymmetrical proteins with a large hydrophilic cytoplasmic domain. PMID- 9726932 TI - Binding of transducin and transducin-derived peptides to rhodopsin studies by attenuated total reflection-Fourier transform infrared difference spectroscopy. AB - Fourier transform infrared difference spectroscopy combined with the attenuated total reflection technique allows the monitoring of the association of transducin with bovine photoreceptor membranes in the dark. Illumination causes infrared absorption changes linked to formation of the light-activated rhodopsin transducin complex. In addition to the spectral changes normally associated with meta II formation, prominent absorption increases occur at 1735 cm-1, 1640 cm-1, 1550 cm-1, and 1517 cm-1. The D2O sensitivity of the broad carbonyl stretching band around 1735 cm-1 indicates that a carboxylic acid group becomes protonated upon formation of the activated complex. Reconstitution of rhodopsin into phosphatidylcholine vesicles has little influence on the spectral properties of the rhodopsin-transducin complex, whereas pH affects the intensity of the carbonyl stretching band. AC-terminal peptide comprising amino acids 340-350 of the transducin alpha-subunit reproduces the frequencies and isotope sensitivities of several of the transducin-induced bands between 1500 and 1800 cm-1, whereas an N-terminal peptide (aa 8-23) does not. Therefore, the transducin-induced absorption changes can be ascribed mainly to an interaction between the transducin-alpha C-terminus and rhodopsin. The 1735 cm-1 vibration is also seen in the complex with C-terminal peptides devoid of free carboxylic acid groups, indicating that the corresponding carbonyl group is located on rhodopsin. PMID- 9726934 TI - Asymmetrical contributions of subunit pore regions to ion selectivity in an inward rectifier K+ channel. AB - We have investigated aspects of ion selectivity in K+ channels by functional expression of wild-type and mutant heteromultimeric G protein-coupled inward rectifier K+ (GIRK) channels in Xenopus oocytes. Within the K+ channel pore (P) region signature sequence, a large number of point mutations in GIRK1 and GIRK4 subunits have been made at a key tyrosine residue--the "signature" tyrosine of the GYG. Studies of mutant GIRK1/GIRK4 heteromultimers reveal that the GIRK1 and GIRK4 subunits contribute asymmetrically to K+ selectivity. The signature tyrosine of GIRK1 can be mutated to many different residues while retaining selectivity; in contrast, the analogous position in GIRK4 must be tyrosine for maximum selectivity. Other residues of the P region also contribute to selectivity, and studies with GIRK1/GIRK4 chimeras reveal that an intact, heteromultimeric P region is necessary and sufficient for optimal K+ selectivity. We propose that the GIRK1 and GIRK4 P regions play roles similar to the two P regions of an emerging family of K+ channels whose subunits each have two P regions connected in tandem. We find different consequences between similar mutations in inward-rectifier and voltage-gated K+ channels, which suggests that the pore structures and selectivity mechanisms in the two classes of channel may not be identical. We confirm that GIRK4 subunits alone can form functional channels in oocytes, but we find that these channels are measurably permeable to Na2+ and Ca2+. PMID- 9726935 TI - Kinetics of Na(+)-dependent conformational changes of rabbit kidney Na+,K(+) ATPase. AB - The kinetics of Na(+)-dependent partial reactions of the Na+,K(+)-ATPase from rabbit kidney were investigated via the stopped-flow technique, using the fluorescent labels N-(4-sulfobutyl)-4-(4-(p-(dipentylamino)phenyl)butadienyl)py ridinium inner salt (RH421) and 5-iodoacetamidofluorescein (5-IAF). When covalently labeled 5-IAF enzyme is mixed with ATP, the two labels give almost identical kinetic responses. Under the chosen experimental conditions two exponential time functions are necessary to fit the data. The dominant fast phase, 1/tau 1 approximately 155 s-1 for 5-IAF-labeled enzyme and 1/tau 1 approximately 200 s-1 for native enzyme (saturating [ATP] and [Na+], pH 7.4 and 24 degrees C), is attributed to phosphorylation of the enzyme and a subsequent conformational change (E1ATP(Na+)3-->E2P(Na+)3 + ADP). The smaller amplitude slow phase, 1/tau 2 = 30-45 s-1, is attributed to the relaxation of the dephosphorylation/rephosphorylation equilibrium in the absence of K+ ions (E2P<==>E2). The Na+ concentration dependence of 1/tau 1 showed half-saturation at a Na+ concentration of 6-8 mM, with positive cooperatively involved in the occupation of the Na+ binding sites. The apparent dissociation constant of the high-affinity ATP-binding site determined from the ATP concentration dependence of 1/tau 1 was 8.0 (+/- 0.7) microM. It was found that P3-1-(2-nitrophenyl)ethyl ATP, tripropylammonium salt (NPE-caged ATP), at concentrations in the hundreds of micromolar range, significantly decreases the value of 1/tau 1, observed. This, as well as the biexponential nature of the kinetic traces, can account for previously reported discrepancies in the rates of the reactions investigated. PMID- 9726937 TI - Molecular ordering of interfacially localized tryptophan analogs in ester- and ether-lipid bilayers studied by 2H-NMR. AB - Perdeuterated indole-d6 and N-methylated indole-d6 were solubilized in lamellar liquid crystalline phases composed of either 1,2-diacyl-glycero-3-phosphocholine (14:0)/water or 1,2-dialkyl-glycero-3-phosphocholine(14:0/water. The molecular ordering of the tryptophan analogs was determined from deuteron quadrupole splittings observed in 2H-NMR spectra on macroscopically aligned lipid bilayers. NMR spectra were recorded with the bilayers oriented perpendicular to or parallel with the external magnetic field, and the values of the splittings differed by a factor of 2 between these distinct orientations, indicating fast rotational motion of the molecules about an axis parallel to the bilayer normal. In all cases the splittings were found to decrease with increasing temperature. Relatively large splittings were observed in all systems, demonstrating that the tryptophans partition into a highly anisotropic environment. Solubilization most likely occurs at the lipid/water interface, as indicated by 1H-NMR chemical shift studies. The 2H-NMR spectra obtained for each analog were found to be rather similar in ester and ether lipids, but with smaller splittings in the ether lipid under similar conditions. The difference was slightly less for the indole molecule. Furthermore, in both lipid systems the positions of the splittings from indole were different from those of N-methyl indole. The results suggest that 1) the tryptophan analogs are solubilized in the interfacial region of the lipid bilayer, 2) the behavior may be modulated by hydrogen bonding in the case of indole, and 3) hydrogen bonding with the lipid carbonyl groups is not likely to play a major role in the solubilization of single indole molecules in the ester lipid bilayer interface. PMID- 9726936 TI - Constitutive secretion of exogenous neurotransmitter by nonneuronal cells: implications for neuronal secretion. AB - Fibroblasts in cell culture were loaded with exogenous neurotransmitter acetylcholine (ACh). ACh secretion from loaded cells was detected by whole-cell patch clamp recordings from Xenopus myocytes manipulated into contact with ACh loaded cells. Two different approaches were used for ACh loading. In the first approach, fibroblasts were incubated in the culture medium containing ACh. Recordings from myocytes revealed fast inward currents that resemble miniature endplate currents found at neuromuscular synapses. The currents observed in recordings from myocytes were due to exocytosis of ACh-containing vesicles. Although exogenous ACh penetrated through the plasma membrane of fibroblasts during incubation and was present in the cytoplasm at detectable levels, cytoplasmic ACh did not contribute to the quantal ACh secretion. In the second approach, exogenous ACh was loaded into the cytoplasm of fibroblasts by microinjection. Under these experimental conditions, fibroblasts also exhibited spontaneous quantal ACh secretion. Analysis of the exocytotic events in fibroblasts following two different protocols of ACh loading revealed that the vesicular compartments responsible for uptake of exogenous ACh are associated with the endocytic recycling pathway. Extrapolation of our results to neuronal cells suggest that in cholinergic neurons, in addition to genuine synaptic vesicles, ACh can be secreted by the vesicles participating in endosomal membrane recycling. PMID- 9726938 TI - A carbon-13 nuclear magnetic resonance spectroscopic study of inter-proton pair order parameters: a new approach to study order and dynamics in phospholipid membrane systems. AB - We report a simple new nuclear magnetic resonance (NMR) spectroscopic method to investigate order and dynamics in phospholipids in which inter-proton pair order parameters are derived by using high resolution 13C cross-polarization/magic angle spinning (CP/MAS) NMR combined with 1H dipolar echo preparation. The resulting two-dimensional NMR spectra permit determination of the motionally averaged interpair second moment for protons attached to each resolved 13C site, from which the corresponding interpair order parameters can be deducted. A spin lock mixing pulse before cross-polarization enables the detection of spin diffusion amongst the different regions of the lipid molecules. The method was applied to a variety of model membrane systems, including 1,2-dimyristoyl-sn glycero-3-phosphocholine (DMPC)/sterol and 1-palmitoyl-2-oleoyl-sn-glycero-3 phosphocholine (POPC)/sterol model membranes. The results agree well with previous studies using specifically deuterium labeled or predeuterated phospholipid molecules. It was also found that efficient spin diffusion takes place within the phospholipid acyl chains, and between the glycerol backbone and choline headgroup of these molecules. The experiment was also applied to biosynthetically 13C-labeled ergosterol incorporated into phosphatidylcholine bilayers. These results indicate highly restricted motions of both the sterol nucleus and the aliphatic side chain, and efficient spin exchange between these structurally dissimilar regions of the sterol molecule. Finally, studies were carried out in the lamellar liquid crystalline (L alpha) and inverted hexagonal (HII) phases of 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE). These results indicated that phosphatidylethanolamine lamellar phases are more ordered than the equivalent phases of phosphatidylcholines. In the HII (inverted hexagonal) phase, despite the increased translational freedom, there is highly constrained packing of the lipid molecules, particularly in the acyl chain region. PMID- 9726939 TI - A mechanism of protein-mediated fusion: coupling between refolding of the influenza hemagglutinin and lipid rearrangements. AB - Although membrane fusion mediated by influenza virus hemagglutinin (HA) is the best characterized example of ubiquitous protein-mediated fusion, it is still not known how the low-pH-induced refolding of HA trimers causes fusion. This refolding involves 1) repositioning of the hydrophobic N-terminal sequence of the HA2 subunit of HA ("fusion peptide"), and 2) the recruitment of additional residues to the alpha-helical coiled coil of a rigid central rod of the trimer. We propose here a mechanism by which these conformational changes can cause local bending of the viral membrane, priming it for fusion. In this model fusion is triggered by incorporation of fusion peptides into viral membrane. Refolding of a central rod exerts forces that pull the fusion peptides, tending to bend the membrane around HA trimer into a saddle-like shape. Elastic energy drives self assembly of these HA-containing membrane elements in the plane of the membrane into a ring-like cluster. Bulging of the viral membrane within such cluster yields a dimple growing toward the bound target membrane. Bending stresses in the lipidic top of the dimple facilitate membrane fusion. We analyze the energetics of this proposed sequence of membrane rearrangements, and demonstrate that this simple mechanism may explain some of the known phenomenological features of fusion. PMID- 9726940 TI - Surface specific kinetics of lipid vesicle adsorption measured with a quartz crystal microbalance. AB - We have measured the kinetics of adsorption of small (12.5-nm radius) unilamellar vesicles onto SiO2, oxidized gold, and a self-assembled monolayer of methyl terminated thiols, using a quartz crystal microbalance (QCM). Simultaneous measurements of the shift in resonant frequency and the change in energy dissipation as a function of time provide a simple way of characterizing the adsorption process. The measured parameters correspond, respectively, to adsorbed mass and to the mechanical properties of the adsorbed layer as it is formed. The adsorption kinetics are surface specific; different surfaces cause monolayer, bilayer, and intact vesicle adsorption. The formation of a lipid bilayer on SiO2 is a two-phase process in which adsorption of a layer of intact vesicles precedes the formation of the bilayer. This is, to our knowledge, the first direct evidence of intact vesicles as a precursor to bilayer formation on a planar substrate. On an oxidized gold surface, the vesicles adsorb intact. The intact adsorption of such small vesicles has not previously been demonstrated. Based on these results, we discuss the capacity of QCM measurements to provide information about the kinetics of formation and the properties of adsorbed layers. PMID- 9726942 TI - Effect of lipid characteristics on the structure of transmembrane proteins. AB - The activity of embedded proteins is known to vary with lipid characteristics. Indeed, it has been shown that some cell-membrane proteins cannot function unless certain non-bilayer-forming lipids (i.e., nonzero spontaneous curvature) are present. In this paper we show that membranes exert a line tension on transmembrane proteins. The line tension, on the order of 1-100 kT/protein, varies with the lipid properties and the protein configuration. Thus, membranes composed of different lipids favor different protein conformations. Model predictions are in excellent agreement with the data of Keller et al. (Biophys. J. 1993, 65:23-27) regarding the conductance of alamethicin channels. PMID- 9726941 TI - The size of the unstirred layer as a function of the solute diffusion coefficient. AB - By monitoring the concentration distribution of several solutes that are diffusing at the same time under given mixing conditions, it was established that the unstirred layer (USL) has no clearly defined boundary. For the cases of solute permeation and water movement across planar bilayer lipid membranes, respectively, experiments carried out with double-barreled microelectrodes have shown that the thickness of the USL depends on which species is diffusing. Small molecules with a larger diffusion coefficient encounter an apparently thicker USL than larger molecules with a smaller diffusion coefficient. The ratio of the USL thicknesses of two different substances is equal to the third root of the ratio of the respective diffusion coefficients. This experimental finding is in good agreement with theoretical predictions from the theory of physicochemical hydrodynamics. PMID- 9726943 TI - Electropermeabilization of mammalian cells to macromolecules: control by pulse duration. AB - Membrane electropermeabilization to small molecules depends on several physical parameters (pulse intensity, number, and duration). In agreement with a previous study quantifying this phenomenon in terms of flow (Rols and Teissie, Biophys. J. 58:1089-1098, 1990), we report here that electric field intensity is the deciding parameter inducing membrane permeabilization and controls the extent of the cell surface where the transfer can take place. An increase in the number of pulses enhances the rate of permeabilization. The pulse duration parameter is shown to be crucial for the penetration of macromolecules into Chinese hamster ovary cells under conditions where cell viability is preserved. Cumulative effects are observed when repeated pulses are applied. At a constant number of pulses/pulse duration product, transfer of molecules is strongly affected by the time between pulses. The resealing process appears to be first-order with a decay time linearly related to the pulse duration. Transfer of macromolecules to the cytoplasm can take place only if they are present during the pulse. No direct transfer is observed with a postpulse addition. The mechanism of transfer of macromolecules into cells by electric field treatment is much more complex than the simple diffusion of small molecules through the electropermeabilized plasma membrane. PMID- 9726944 TI - The stiffness of rabbit skeletal actomyosin cross-bridges determined with an optical tweezers transducer. AB - Muscle contraction is brought about by the cyclical interaction of myosin with actin coupled to the breakdown of ATP. The current view of the mechanism is that the bound actomyosin complex (or "cross-bridge") produces force and movement by a change in conformation. This process is known as the "working stroke." We have measured the stiffness and working stroke of a single cross-bridge (kappa xb, dxb, respectively) with an optical tweezers transducer. Measurements were made with the "three bead" geometry devised by Finer et al. (1994), in which two beads, supported in optical traps, are used to hold an actin filament in the vicinity of a myosin molecule, which is immobilized on the surface of a third bead. The movements and forces produced by actomyosin interactions were measured by detecting the position of both trapped beads. We measured, and corrected for, series compliance in the system, which otherwise introduces large errors. First, we used video image analysis to measure the long-range, force-extension property of the actin-to-bead connection (kappa con), which is the main source of "end compliance." We found that force-extension diagrams were nonlinear and rather variable between preparations, i.e., end compliance depended not only upon the starting tension, but also upon the F-actin-bead pair used. Second, we measured kappa xb and kappa con during a single cross-bridge attachment by driving one optical tweezer with a sinusoidal oscillation while measuring the position of both beads. In this way, the bead held in the driven optical tweezer applied force to the cross-bridge, and the motion of the other bead measured cross-bridge movement. Under our experimental conditions (at approximately 2 pN of pretension), connection stiffness (kappa con) was 0.26 +/- 0.16 pN nm-1. We found that rabbit heavy meromyosin produced a working stroke of 5.5 nm, and cross bridge stiffness (kappa xb) was 0.69 +/- 0.47 pN nm-1. PMID- 9726945 TI - Spontaneous oscillatory contraction without regulatory proteins in actin filament reconstituted fibers. AB - Skinned skeletal and cardiac muscle fibers exhibits spontaneous oscillatory contraction (SPOC) in the presence of MgATP, MgADP, and inorganic phosphate (Pi)1 but the molecular mechanism underlying this phenomenon is not yet clear. We have investigated the role of regulatory proteins in SPOC using cardiac muscle fibers of which the actin filaments had been reconstituted without tropomyosin and troponin, according to a previously reported method (Fujita et al., 1996. Biophys. J. 71:2307-2318). That is, thin filaments in glycerinated cardiac muscle fibers were selectively removed by treatment with gelsolin. Then, by adding exogenous actin to these thin filament-free cardiac muscle fibers under polymerizing conditions, actin filaments were reconstituted. The actin filament reconstituted cardiac muscle fibers generated active tension in a Ca(2+) insensitive manner because of the lack of regulatory proteins. Herein we have developed a new solvent condition under which SPOC occurs, even in actin filament reconstituted fibers: the coexistence of 2,3-butanedione 2-monoxime (BDM), a reversible inhibitor of actomyosin interactions, with MgATP, MgADP and Pi. The role of BDM in the mechanism of SPOC in the actin filament-reconstituted fibers was analogous to that of the inhibitory function of the tropomyosin-troponin complex (-Ca2+) in the control fibers. The present results suggest that SPOC is a phenomenon that is intrinsic to the actomyosin motor itself. PMID- 9726946 TI - Structural characterization of the L-to-M transition of the bacteriorhodopsin photocycle. AB - Structural intermediates occurring in the photocycle of wild-type bacteriorhodopsin are trapped by illuminating hydrated, glucose-embedded purple membrane at 170 K, 220 K, 230 K, and 240 K. We characterize light-induced changes in protein conformation by electron diffraction difference Fourier maps, and relate these to previous work on photocycle intermediates by infrared (FTIR) spectroscopy. Samples illuminated at 170 K are confirmed by FTIR spectroscopy to be in the L state; a difference Fourier projection map shows no structural change within the 0.35-nm resolution limit of our data. Difference maps obtained with samples illuminated at 220 K, 230 K, and 240 K, respectively, reveal a progressively larger structural response in helix F when the protein is still in the M state, as judged by the FTIR spectra. Consistent with previous structural studies, an adjustment in the position or in the degree of ordering of helix G accompanies this motion. The model of the photocycle emerging from this and previous studies is that bacteriorhodopsin experiences minimal change in protein structure until a proton is transferred from the Schiff base to Asp85. The M intermediate then undergoes a conformational evolution that opens a hydrated "half-channel," allowing the subsequent reprotonation of the Schiff base by Asp96. PMID- 9726948 TI - Photoresponses of Halobacterium salinarum to repetitive pulse stimuli. AB - Halobacterium salinarum cells from 3-day-old cultures have been stimulated with different patterns of repetitive pulse stimuli. A short train of 0.6-s orange light pulses with a 4-s period resulted in reversal peaks of increasing intensity. The reverse occurred when blue light pulses were delivered as a finite train: with a 3-s period, the response declined in sequence from the first to the last pulse. To evaluate the response of the system under steady-state conditions of stimulation, continuous trains of pulses were also applied; whereas blue light always produced a sharply peaked response immediately after each pulse, orange pulses resulted in a declining peak of reversals that lasted until the subsequent pulse. An attempt to account for these results in terms of current excitation/adaptation models shows that additional mechanisms appear to be at work in this transduction chain. PMID- 9726947 TI - Connectivity of the retinal Schiff base to Asp85 and Asp96 during the bacteriorhodopsin photocycle: the local-access model. AB - In the recently proposed local-access model for proton transfers in the bacteriorhodopsin transport cycle (Brown et al. 1998. Biochemistry. 37:3982 3993), connection between the retinal Schiff base and Asp85 (in the extracellular direction) and Asp96 (in the cytoplasmic direction)is maintained as long as the retinal is in its photoisomerized state. The directionality of the proton translocation is determined by influences in the protein that make Asp85 a proton acceptor and, subsequently, Asp96 a proton donor. The idea of concurrent local access of the Schiff base in the two directions is now put to a test in the photocycle of the D115N/D96N mutant. The kinetics had suggested that there is a single sequence of intermediates, L<-->M1<-->M2<-->N, and the M2-->M1 reaction depends on whether a proton is released to the extracellular surface. This is now confirmed. We find that at pH 5, where proton release does not occur, but not at higher pH, the photostationary state created by illumination with yellow light contains not only the M1 and M2 states, but also the L and the N intermediates. Because the L and M1 states decay rapidly, they can be present only if they are in equilibrium with later intermediates of the photocycle. Perturbation of this mixture with a blue flash caused depletion of the M intermediate, followed by its partial recovery at the expense of the L state. The change in the amplitude of the C=O stretch band at 1759 cm-1 demonstrated protonation of Asp85 in this process. Thus, during the reequilibration the Schiff base lost its proton to Asp85. Because the N state, also present in the mixture, arises by protonation of the Schiff base from the cytoplasmic surface, these results fulfill the expectation that under the conditions tested the extracellular access of the Schiff base would not be lost at the time when there is access in the cytoplasmic direction. Instead, the connectivity of the Schiff base flickers rapidly (with the time constant of the M1<-->M2 equilibration) between the two directions during the entire L-to-N segment of the photocycle. PMID- 9726950 TI - Ionic strength-dependent physicochemical factors in cytochrome c3 regulating the electron transfer rate. AB - The effect of ionic strength on the macroscopic and microscopic redox potentials and the heme environment of cytochrome c3 from Desulfovibrio vulgaris Miyazaki F have been investigated by NMR and electrochemical methods. The redox potentials of this tetraheme protein are found to be ionic strength-dependent. Especially, the microscopic redox potentials of hemes 2 and 3 at the fourth reduction step increase significantly with increasing ionic strength, which is in contraction to the theoretical expectation. The coordinated imidazole proton signals are unaffected by ionic strength. However, the methyl and propionate proton signals of hemes 1 and 4 showed significant ionic strength dependencies that are distinct from those for hemes 2 and 3. This heme classification is the same as that found in the ionic strength dependencies of the microscopic redox potentials at the fourth reduction step. Furthermore, the effect of ionic strength on the electrostatic potentials at the heme irons has been examined on the theoretical basis. The electrostatic potential at heme 4 changes up to 1 M ionic strength, which was not expected from the observations reported on cytochromes so far. These results are discussed in connection with the reported anomalous ionic strength dependency of the reduction rate of cytochrome c3. PMID- 9726949 TI - Contributions of a highly conserved VH/VL hydrogen bonding interaction to scFv folding stability and refolding efficiency. AB - The assembly of single-chain Fv (scFv) antibody fragments, consisting of an interconnected variable heavy chain (VH) and variable light chain (VL), is a cooperative process that requires coupled folding and domain association. We report here an initial investigation of VH/VL domain-domain assembly with a site directed mutagenesis study that probes a highly conserved VH/VL hydrogen bonding interaction. Gln168 of the S5 scFv (Kabat VH 39) is absolutely conserved in 95% of all VH, and Gln44 (Kabat VL 38) is found in 94% of all kappa VL (Glx in 95% of all lambda VL). These side chains form two hydrogen bonds in head-to-tail alignment across the VH/VL interface. Double mutant cycles at Gln168 and Gln44 were constructed to first investigate their contribution to thermodynamic folding stability, second to investigate whether stability can be improved, and third to determine whether refolding efficiencies are affected by mutations at these positions. The results demonstrate that the Gln168-Gln44 interaction is not a key determinant of S5 scFv folding stability, as sequential modification to alanine has no significant effect on the free energy of folding. Several mutations that alter the glutamines to methionine or charged amino acids significantly increase the thermodynamic stability by increasing the m(g) associated with the unfolding isotherm. These effects are hypothesized to arise largely from an increase in the VH/VL association free energy that leads to tighter coupling between domain domain association and folding. All of the mutants also display a reduced antigen binding affinity. Single and double methionine mutants also displayed significant increases in refolding efficiency of 2.4- to 3-fold over the native scFv, whereas the double alanine/methionine mutants displayed moderate 1.9- to 2.4-fold enhancement. The results suggest that reengineering the VH/VL interface could be useful in improving the stability of single-chain antibodies, as Ala/Met mutations at these conserved positions increase the free energy of folding by 46% while minimally perturbing binding affinity. They also could be useful in improving scFv recovery from inclusion bodies as the mutations increase the refolding efficiency by more than twofold. PMID- 9726952 TI - Sedimentation analysis of noninteracting and self-associating solutes using numerical solutions to the Lamm equation. AB - The potential of using the Lamm equation in the analysis of hydrodynamic shape and gross conformation of proteins and reversibly formed protein complexes from analytical ultracentrifugation data was investigated. An efficient numerical solution of the Lamm equation for noninteracting and rapidly self-associating proteins by using combined finite-element and moving grid techniques is described. It has been implemented for noninteracting solutes and monomer-dimer and monomer-trimer equilibria. To predict its utility, the error surface of a nonlinear regression of simulated sedimentation profiles was explored. Error contour maps were calculated for conventional independent and global analyses of experiments with noninteracting solutes and with monomer-dimer systems at different solution column heights, loading concentrations, and centrifugal fields. It was found that the rotor speed is the major determinant for the shape of the error surface, and that global analysis of different experiments can allow substantially improved characterization of the solutes. We suggest that the global analysis of the approach to equilibrium in a short-column sedimentation equilibrium experiment followed by a high-speed short-column sedimentation velocity experiment can result in sedimentation and diffusion coefficients of very high statistical accuracy. In addition, in the case of a protein in rapid monomer-dimer equilibrium, this configuration was found to reveal the most precise estimate of the association constant. PMID- 9726951 TI - The photoexcited triplet state as a probe of chromophore-protein interaction in myoglobin. AB - The photoexcited metastable triplet state of Mg(2+)-mesoporphyrin IX (MgMPIX) or Mg(2+)-protoporphyrin IX (MgPPIX) located in the heme pocket of horse myoglobin (Mb) was investigated by optical and electron paramagnetic resonance (EPR) spectroscopy, and its properties were compared with the model complexes, MgMPIX, MgPPIX, and Mg2+ etioporphyrin I (MgETIOI), in noncoordinating and coordinating organic glasses. Zero-field splitting parameters, line shape, and Jahn-Teller distortion in the temperature range of 3.8-110 K are discussed in terms of porphyrin-protein interactions. The triplet line shapes for MgMPIXMb and MGPPIXMb show no temperature-dependent spectral line shape changes suggestive of Jahn Teller dynamics, and it is concluded that the energy splitting is >> 150 cm-1, suggesting symmetry breaking from the anisotropy of intermal electric fields of the protein, and consistent with previous predictions (Geissinger et al. 1995. J. Phys. Chem. 99:16527-16529). Both MgMPIXMb and MgPPIXMb demonstrate electron spin polarization at low temperature, and from the polarization pattern it can be concluded that intersystem crossing occurs predominantly into in-plane spin sublevels of the triplet state. The splitting in the Q0.0 absorption band and the temperature dependence and splitting of the photoexcited triplet state of myoglobin in which the iron was replaced by Mg2+ are interpreted in terms of effects produced by electric field asymmetry in the heme pocket. PMID- 9726953 TI - Continuous wave two-photon scanning near-field optical microscopy. AB - We have implemented continuous-wave two-photon excitation of near-UV absorbing fluorophores in a scanning near-field optical microscope (SNOM). The 647-nm emission of an Ar-Kr mixed gas laser was used to excite the UV-absorbing DNA dyes DAPI, the bisbenzimidazole Hoechst 33342, and ethidium bromide in a shared aperture SNOM with uncoated fiber tips. Polytene chromosomes of Drosophila melanogaster and the nuclei of 3T3 Balb/c cells labeled with these dyes were readily imaged. The fluorescence intensity showed the expected nonlinear (second order) dependence on the excitation power in the range of 8-180 mW. We measured the fluorescence intensity as a function of the tip-sample displacement in the direction normal to the sample surface in the single- and two-photon excitation modes (SPE, TPE). The fluorescence intensity decayed faster in TPE than in SPE. PMID- 9726954 TI - Arginine activates glycolysis of goat epididymal spermatozoa: an NMR study. AB - The present study explores the mechanism underlying the action of L-arginine on the metabolic activity of spermatozoa. Goat epididymal spermatozoa were incubated with different concentrations of L-arginine to determine its effect on the utilization of glucose, fructose, and pyruvate. NMR techniques have been applied to elucidate the effect of L-arginine, L-lysine, and L-ornithine on the glycolysis of epididymal goat spermatozoa. Whereas 31P NMR has been used to estimate the change of pH in the presence of different concentrations of L arginine, 13C NMR has been used to estimate the substrate consumption and lactate production. At optimal concentration of L-arginine, the forward metabolic rates have been found to increase by two to three times over control experiments. Arginine is not consumed in these reactions, but acts as an activator. Longitudinal relaxation time (T1) measurements indicate that the guanidino group of L-arginine plays an active role in binding to cells. The amino acid L-lysine is less effective, and L-ornithine is ineffective. PMID- 9726955 TI - Simulating the role of microtubules in depolymerization-driven transport: a Monte Carlo approach. AB - In this paper we present a model that simulates the role of microtubules in depolymerization-driven transport. The model simulates a system that consists of a 13-protofilament microtubule with "five-start" helical structure and a motor protein-coated bead that moves along one of the protofilaments of the microtubule, as in in vitro experiments. The microtubule is simulated using the lateral cap model, with substantial generalizations. For the new terminal configurations in the presence of the bead, rate constants for association and dissociation events of tubulin molecules are calculated by exploring the geometric similarities between different patterns of terminal configurations and by decomposing complex patterns into simpler patterns whose corresponding rate constants are known. In comparison with a previous model, in which simplifications are made about the structure of the microtubule and in which the microtubule can only depolymerize, the detailed structure of the microtubule is taken into account in the present model. Furthermore, the microtubule can be either polymerizing or depolymerizing. Force-velocity curves are obtained for both zero and non-zero tubulin guanosine 5'-triphosphate (GTP) concentrations. By analyzing the trajectory of the bead under different parameters, the condition for "run and pause" is analyzed, and the time scale of "run" and "pause" is found to be different for different motor proteins. We also suggest experiments that can be used to examine the results predicted by the model. PMID- 9726956 TI - Cell transit analysis of ligand-induced stiffening of polymorphonuclear leukocytes. AB - A mathematical treatment of the mechanical behavior of transiently bonded polymer networks is used to interpret measurements of the pressure-induced passage of plant cells through microporous membranes. Cell transit times are inferred to be proportional to the instantaneous shear modulus of the cell cortex, a parameters that we then relate to properties of the cortical F-actin matrix. These theoretical results are used to analyze published data on chemoattractant-induced changes of rigidity of polymorphonuclear leukocytes. We thereby rationalize previously noted, peculiar, power-law logarithmic dependences of transit time on ligand concentration. As a consequence, we are able to deduce a linear relationship between the extent of F-actin polymerization and the logarithm of the chemoattractant concentration. The latter is examined with regard to the G protein activation that is known to occur when chemoattractants bind to receptors on the surfaces of polymorphonuclear cells. PMID- 9726957 TI - Measuring two-dimensional receptor-ligand binding kinetics by micropipette. AB - We report a novel method for measuring forward and reverse kinetic rate constants, kf0 and kr0, for the binding of individual receptors and ligands anchored to apposing surfaces in cell adhesion. Not only does the method examine adhesion between a single pair of cells; it also probes predominantly a single receptor-ligand bond. The idea is to quantify the dependence of adhesion probability on contact duration and densities of the receptors and ligands. The experiment was an extension of existing micropipette protocols. The analysis was based on analytical solutions to the probabilistic formulation of kinetics for small systems. This method was applied to examine the interaction between Fc gamma receptor IIIA (CD16A) expressed on Chinese hamster ovary cell transfectants and immunoglobulin G (IgG) of either human or rabbit origin coated on human erythrocytes, which were found to follow a monovalent biomolecular binding mechanism. The measured rate constants are Ackf0 = (2.6 +/- 0.32) x 10(-7) micron 4 s-1 and kr0 = (0.37 +/- 0.055) s-1 for the CD16A-hIgG interaction and Ackf0 = (5.7 +/- 0.31) X 10(-7) micron 4 s-1 and kr0 = (0.20 +/- 0.042) s-1 for the CD16A rIgG interaction, respectively, where Ac is the contact area, estimated to be a few percent of 3 micron 2. PMID- 9726958 TI - Simulations of the erythrocyte cytoskeleton at large deformation. I. Microscopic models. AB - Three variations of a polymer chain model for the human erythrocyte cytoskeleton are used in large deformation simulations of microscopic membrane patches. Each model satisfies an experimental observation that the contour length of the spectrin tetramers making up the erythrocyte cytoskeleton is roughly square root of 7 times the end-to-end distance of the tetramer in vivo. Up to modest stress, each brushy cytoskeletal network behaves, consistently, like a low-temperature, planar network of Hookean springs, with a model-dependent effective spring constant, keff, in the range of 20-40 kBT/s(o)2, where T is the temperature and s(o) is the force-free spring length. However, several features observed at large deformation distinguish these models from spring networks: 1) Network dimensions do not expand without bound in approaching a critical isotropic tension (square root of 3 keff) that is a characteristic limit of Hookean spring nets. 2) In surface compression, steric interactions among the chain elements prevent a network collapse that is otherwise observed in compression of planar triangulated networks of springs. 3) Under uniaxial surface tension, isotropy of the network disappears only as the network is stretched by more than 50% of its equilibrium dimensions. Also found are definitively non-Hookean regimes in the stress dependence of the elastic moduli. Lastly, determinations of elastic moduli from both fluctuations and stress/strain relations prove to be consistent, implying that consistency should be expected among experimental determinations of these quantities. PMID- 9726959 TI - Simulations of the erythrocyte cytoskeleton at large deformation. II. Micropipette aspiration. AB - Coarse-grained molecular models of the erythrocyte membrane's spectrin cytoskeleton are presented in Monte Carlo simulations of whole cells in micropipette aspiration. The nonlinear chain elasticity and sterics revealed in more microscopic cytoskeleton models (developed in a companion paper; Boey et al., 1998. Biophys. J. 75:1573-1583) are faithfully represented here by two- and three-body effective potentials. The number of degrees of freedom of the system are thereby reduced to a range that is computationally tractable. Three effective models for the triangulated cytoskeleton are developed: two models in which the cytoskeleton is stress-free and does or does not have internal attractive interactions, and a third model in which the cytoskeleton is prestressed in situ. These are employed in direct, finite-temperature simulations of erythrocyte deformation in a micropipette. All three models show reasonable agreement with aspiration measurements made on flaccid human erythrocytes, but the prestressed model alone yields optimal agreement with fluorescence imaging experiments. Ensemble-averaging of nonaxisymmetrical, deformed structures exhibiting anisotropic strain are thus shown to provide an answer to the basic question of how a triangulated mesh such as that of the red cell cytoskeleton deforms in experiment. PMID- 9726960 TI - Glycosylphosphatidylinositol-anchored and secretory isoforms of mono-ADP ribosyltransferases. PMID- 9726961 TI - Activation of caspase-1 in the nucleus requires nuclear translocation of pro caspase-1 mediated by its prodomain. AB - The interleukin-1beta-converting enzyme-like protease precursor, pro-caspase-1, has an N-terminal prodomain that is removed during cleavage activation of the protease. Here we show that tumor necrosis factor treatment of HeLa cells induced apoptosis without detectable proteolytic activation of caspase-1 in the cytosol. Instead, tumor necrosis factor induced the translocation of pro-caspase-1 to the nucleus where it was proteolytically activated, releasing the intact prodomain. We identified a nuclear localization signal in the prodomain, which was required for translocation of both pro-caspase-1 as well as its prodomain to the nucleus. Surprisingly, transfected MCF-7 carcinoma or embryonic kidney 293T cells expressing the prodomain alone underwent apoptosis. These results show that death signal-induced nuclear targeting is a novel activity of a caspase prodomain and indicate that caspase-1 and its prodomain may have hitherto unsuspected nuclear functions in apoptosis. PMID- 9726962 TI - The copper chaperone CCS directly interacts with copper/zinc superoxide dismutase. AB - Dominantly inherited mutations in the gene encoding copper/zinc superoxide dismutase (SOD1) result in the fatal motor neuron disease familial amyotrophic lateral sclerosis (FALS). These mutations confer a gain-of-function to SOD1 with neuronal degeneration resulting from enhanced free radical generating activity of the copper present in the mutant enzyme. The delivery of copper to SOD1 is mediated through a soluble factor identified as the copper chaperone for SOD1 (CCS). Amino acid sequence alignment of SOD1 and CCS reveals a striking homology with conservation of the amino acids essential for mediating SOD1 homodimerization. Here we demonstrate that CCS and SOD1 directly interact in vitro and in vivo and that this interaction is mediated via the homologous domains in each protein. Importantly, CCS interacts not only with wild-type SOD1 but also with SOD1 containing the common missense mutations resulting in FALS. Our findings therefore reveal a common mechanism whereby different SOD1 FALS mutants may result in neuronal injury and suggest a novel therapeutic approach in patients affected by this fatal disease. PMID- 9726963 TI - Expression cloning and characterization of a transporter for large neutral amino acids activated by the heavy chain of 4F2 antigen (CD98). AB - A cDNA was isolated from rat C6 glioma cells by expression cloning which encodes a novel Na+-independent neutral amino acid transporter designated LAT1. For functional expression in Xenopus oocytes, LAT1 required the heavy chain of 4F2 cell surface antigen (CD98), a type II membrane glycoprotein. When co-expressed with 4F2 heavy chain, LAT1 transported neutral amino acids with branched or aromatic side chains and did not accept basic amino acids or acidic amino acids. The transport via LAT1 was Na+-independent and sensitive to a system L-specific inhibitor 2-aminobicyclo-(2,2,1)-heptane-2-carboxylic acid. These functional properties correspond to those of the classically characterized amino acid transport system L, a major nutrient transporter. In in vitro translation, LAT1 was shown to be a nonglycosylated membrane protein consistent with the property of 4F2 light chain, suggesting LAT1 is at least one of the proteins formerly referred to as 4F2 light chain. LAT1 exhibits relatively low but significant amino acid sequence similarity to mammalian cationic amino acid transporters and amino acid permeases of bacteria and yeasts, indicating LAT1 is a new member of the APC superfamily. Because of highly regulated nature and high level of expression in tumor cell lines, LAT1 is thought to be up-regulated to support the high protein synthesis for cell growth and cell activation. The cloning of LAT1 is expected to facilitate the research on the protein-protein interaction in the transporter field and to provide a clue to the search for still unidentified transporters. PMID- 9726964 TI - A novel regulator of p21-activated kinases. AB - Proteins of the p21-activated kinase (Pak) family have been implicated in the regulation of gene expression, cytoskeletal architecture, and apoptosis. Although the ability of Cdc42 and Rac GTPases to activate Pak is well established, relatively little else is known about Pak regulation or the identity of Pak cellular targets. Here we report the identification of two closely related Pak3 binding proteins, possibly arising from alternative splicing, designated p50 and p85(Cool-1) (cloned out of library). Both isoforms of Cool contain a Src homology 3 domain that directly mediates interaction with Pak3 and tandem Dbl homology and pleckstrin homology domains. Despite the presence of the Dbl homology-pleckstrin homology motif, a characteristic of Rho family activators, activation of Cdc42 or Rac by Cool is not detectable. Instead binding of p50(Cool-1), but not p85(Cool 1), to Pak3 represses its activation by upstream activators such as the Dbl oncoprotein, indicating a novel mechanism of regulation of Pak signaling. PMID- 9726965 TI - Loops and bulge/loops in iron-responsive element isoforms influence iron regulatory protein binding. Fine-tuning of mRNA regulation? AB - A family of noncoding mRNA sequences, iron-responsive elements (IREs), coordinately regulate several mRNAs through binding a family of mRNA-specific proteins, iron regulatory proteins (IRPs). IREs are hairpins with a constant terminal loop and base-paired stems interrupted by an internal loop/bulge (in ferritin mRNA) or a C-bulge (in m-aconitase, erythroid aminolevulinate synthase, and transferrin receptor mRNAs). IRP2 binding requires the conserved C-G base pair in the terminal loop, whereas IRP1 binding occurs with the C-G or engineered U-A. Here we show the contribution of the IRE internal loop/bulge to IRP2 binding by comparing natural and engineered IRE variants. Conversion of the internal loop/bulge in the ferritin-IRE to a C-bulge, by deletion of U, decreased IRP2 binding by >95%, whereas IRP1 binding changed only 13%. Moreover, IRP2 binding to natural IREs with the C-bulge was similar to the DeltaU6 ferritin-IRE: >90% lower than the ferritin-IRE. The results predict mRNA-specific variation in IRE dependent regulation in vivo and may relate to previously observed differences in iron-induced ferritin and m-aconitase synthesis in liver and cultured cells. Variations in IRE structure and cellular IRP1/IRP2 ratios can provide a range of finely tuned, mRNA-specific responses to the same (iron) signal. PMID- 9726967 TI - Uncoupling hydrophobicity and helicity in transmembrane segments. Alpha-helical propensities of the amino acids in non-polar environments. AB - Although the chains of amino acids in proteins that span the membrane are demonstrably helical and hydrophobic, little attention has been paid toward addressing the range of helical propensities of individual amino acids in the non polar environment of membranes. Because it is inappropriate to apply soluble protein-based structure prediction algorithms to membrane proteins, we have used de novo designed peptides (KKAAAXAAAAAXAAWAAXAAAKKKK-amide, where X indicates one of the 20 commonly occurring amino acids) that mimic a protein membrane-spanning domain to determine the alpha-helical proclivity of each residue in the isotropic non-polar environment of n-butanol. Peptide helicities measured by circular dichroism spectroscopy were found to range from theta222 = -17,000 degrees (Pro) to -38,800 degrees (Ile) in n-butanol. The relative helicity of each amino acid is shown to be well correlated with its occurrence frequency in natural transmembrane segments, indicating that the helical propensity of individual residues in concert with their hydrophobicity may be a key determinant of the conformations of protein segments in membranes. PMID- 9726966 TI - Subunits of the yeast SWI/SNF complex are members of the actin-related protein (ARP) family. AB - The yeast SWI/SNF chromatin remodeling complex is comprised of 11 tightly associated polypeptides (SWI1, SWI2, SWI3, SNF5, SNF6, SNF11, SWP82, SWP73, SWP59, SWP61, and SWP29). We have used matrix-assisted laser desorption ionization time-of-flight mass spectrometry to identify the genes that encode the SWP59 and SWP61 subunits. Surprisingly, we find that SWP59 and SWP61 are encoded by the ARP9 and ARP7 genes, respectively, which encode members of the actin related protein (ARP) family. Sequence analyses have shown that ARP9 and ARP7 are 24-26% identical (48-51% similar) to yeast actin and that they are likely to maintain the overall actin fold. Deletion of either the ARP9 or ARP7 gene causes typical swi/snf phenotypes, including growth defects on media containing galactose, glycerol, or sucrose as sole carbon sources. ARP9 and ARP7 are also required for expression of an HO-lacZ fusion gene and for full transcriptional enhancement by the GAL4 activator. The identification of two ARP family members as crucial subunits of the SWI/SNF complex suggests that the complex may contain a total of three different ATPase subunits; furthermore, the similarity of ARP7 and ARP9 to the HSP and HSC family of ATPases suggests the possibility that chromatin remodeling by SWI/SNF may involve chaperone-like activities. PMID- 9726968 TI - Expression of constitutively activated Gialpha2 in vivo ameliorates streptozotocin-induced diabetes. AB - Streptozotocin treatment in vivo generates a model of insulin-dependent diabetes mellitus via destruction of the pancreatic beta-cells responsible for insulin secretion. Tissue-specific expression of the Q205L constitutively activated mutant form of the G-protein Gialpha2 in vivo ameliorates streptozotocin-induced insulin-dependent diabetes mellitus in transgenic mice. Conditional expression of Q205L Gialpha2 in vivo in liver, skeletal muscle, and adipose tissue markedly rectifies glucose tolerance, fasting glucose levels, and glycogen synthase activation in the mice with insulin-dependent diabetes mellitus, providing a novel therapeutic target for diabetes. PMID- 9726969 TI - Solution structure and internal motion of a bioactive peptide derived from nerve growth factor. AB - The conformation and internal dynamics of a bioactive cyclic peptide, N-acetyl YCTDEKQCY, derived from the C-D loop of beta-nerve growth factor (beta-NGF) were analyzed by solution NMR spectroscopy. NMR experimental data were used to calculate an ensemble of peptide structures. All of the structures had a beta turn at residues Asp4-Gln7 but could be divided into two families according the presence or absence of a hydrogen bond at Gln7. Comparison of the calculated structures with the corresponding C-D loops from the x-ray structures of the NGF revealed striking similarity. The orientation of Glu5, Lys6, and Gln7 side chains in the NGF mimetic was very similar to the C-D loop of NGF. These residues are known to participate in interactions with the TrkA receptor. Relaxation measurements of the peptidomimetic alpha-carbons at 13C natural abundance and calculated dynamic parameters suggest that the loop region of peptide is well structured but that residues Thr3, Asp4, Glu5, and Lys6 undergo slow conformational exchange. These results suggest that conformational similarity and possibly peptide dynamics are responsible for the bioactivity of the peptide. PMID- 9726970 TI - p130, p107, and pRb are differentially regulated in proliferating cells and during cell cycle arrest by alpha-interferon. AB - We have determined how the phosphorylation of the retinoblastoma family (pRb, p107, and p130) is governed in individual cell cycle phases of Daudi B-cells during cell cycle exit triggered by alpha-interferon (alpha-IFN). alpha-IFN causes dephosphorylation of pRb and loss of p130 phosphorylated Form 3. However, the change in p130 phosphorylation in response to alpha-IFN occurs before dephosphorylation of pRb is complete because loss of p130 Form 3 occurs throughout the cell cycle prior to complete arrest in G1, whereas pRb is dephosphorylated only in G1. In contrast, p107 is dephosphorylated and is then depleted from cells as they exit the cell cycle. p130, predominantly in Form 1, and hypophosphorylated pRb bind an E2F DNA binding site; p130 complexes E2F-4, whereas pRb binds both E2F-4 and E2F-1. The phosphorylated forms of E2F-4 that bind to the E2F DNA site are different from hyperphosphorylated E2F-4, which predominates in primary hemopoietic cells in G0. We conclude that although cell cycle arrest induced by alpha-IFN may be mediated in part by formation of a complex containing p130 and E2F-4, alpha-IFN does not induce hyperphosphorylation of E2F-4, which characterizes primary hemopoietic cells in G0. PMID- 9726971 TI - An ectoprotein kinase of group C streptococci binds hyaluronan and regulates capsule formation. AB - A 56-kDa protein had been isolated and cloned from protoplast membranes of group C streptococci that had erroneously been identified as hyaluronan synthase. The function of this protein was reexamined. When streptococcal membranes were separated on an SDS-polyacrylamide gel and renatured, a 56-kDa protein was detected that had kinase activity for a casein substrate. When this recombinant protein was expressed in Escherichia coli and incubated in the presence of [32P]ATP, it was responsible for phosphorylation of two proteins with 30 and 56 kDa that were not present in the control lysate. The 56-kDa protein was specifically phosphorylated in an immunoprecipitate of a detergent extract of the recombinant E. coli lysate with antibodies against the 56-kDa protein, indicating that it was autophosphorylated. The E. coli lysate containing the recombinant protein could bind hyaluronan, and hyaluronan binding was abolished by the addition of ATP. Kinetic analysis of hyaluronan synthesis and release from isolated protoplast membranes indicated that phosphorylation by ATP stimulated hyaluronan release and synthesis. Incubation of membranes with antibodies to the 56-kDa protein increased hyaluronan release. The addition of [32P]ATP to intact streptococci led to rapid phosphorylation of two proteins, 56 and 75 kDa each at threonine residues. This phosphorylation was neither observed with [32P]phosphate nor in the presence of trypsin, indicating that the kinase was localized extracellularly. The addition of ATP to growing group C streptococci led to increased hyaluronan synthesis and release. However marked differences were found between group A and group C streptococci. Antibodies against the 56-kDa protein from group C streptococci did not recognize proteins from group A strains, and a homologous DNA sequence could not be detected by polymerase chain reaction or Southern blotting. In addition, Group A streptococci did not retain a large hyaluronan capsule like group C strains. These results indicated that the 56-kDa protein is an ectoprotein kinase specific for group C streptococci that regulates hyaluronan capsule shedding by phosphorylation. PMID- 9726972 TI - Calnexin association is not sufficient to protect T cell receptor alpha proteins from rapid degradation in CD4+CD8+ thymocytes. AB - During T cell development, assembly of the mutisubunit T cell receptor (TCR) complex is regulated by the differential stability of newly synthesized TCRalpha molecules, having a half-life of approximately 20 min in immature CD4+CD8+ thymocytes compared with >75 min in mature T cells. The molecular basis for TCRalpha instability in CD4+CD8+ thymocytes is unknown but has been postulated to involve abnormalities in N-glycan processing and calnexin assembly as perturbation of these pathways markedly destabilizes TCRalpha proteins in all other T cell types examined. Here, we compared the processing of TCRalpha glycoproteins and their assembly with calnexin and calreticulin chaperones in CD4+CD8+ thymocytes and splenic T cells. These studies show that TCRalpha glycoproteins synthesized in CD4+CD8+ thymocytes were processed in a similar manner as those made in splenic T cells and that TCRalpha proteins stably associated with calnexin in both cell types. Interestingly, however, TCRalpha association with the calnexin-related molecule calreticulin was decreased in CD4+CD8+ thymocytes compared with splenic T cells. Finally, TCRalpha degradation in CD4+CD8+ thymocytes was impaired by inhibitors of proteasome activity, which was correlated with stabilization of calnexin.TCRalpha complexes. These data demonstrate that calnexin association is not sufficient to protect TCRalpha proteins from rapid degradation in CD4+CD8+ thymocytes, suggesting that additional components of the quality control system of the endoplasmic reticulum operate to ensure the proper folding of nascent TCRalpha glycoproteins. PMID- 9726974 TI - The role of 3'-5' exonucleolytic proofreading and mismatch repair in yeast mitochondrial DNA error avoidance. AB - In the D171G/D230A mutant generated at conserved aspartate residues in the Exo1 and Exo2 sites of the 3'-5' exonuclease domain of the yeast mitochondrial DNA (mtDNA) polymerase (pol-gamma), the mitochondrial genome is unstable and the frequency of mtDNA point mutations is 1500 times higher than in the wild-type strain and 10 times higher than in single substitution mutants. The 10(4)-fold decrease in the 3'-5' exonuclease activity of the purified mtDNA polymerase is associated with mismatch extension and high rates of base misincorporation. Processivity of the purified polymerase on primed single-stranded DNA is decreased and the Km for dNTP is increased. The sequencing of mtDNA point mutations in the wild-type strain and in proofreading and mismatch-repair deficient mutants shows that mismatch repair contributes to elimination of the transitions while exonucleolytic proofreading preferentially repairs transversions, and more specifically A to T (or T to A) transversions. However, even in the wild-type strain, A to T (or T to A) transversions are the most frequent substitutions, suggesting that they are imperfectly repaired. The combination of both mismatch repair and proofreading deficiencies elicits a mitochondrial error catastrophe. These data show that the faithful replication of yeast mtDNA requires both exonucleolytic proofreading and mismatch repair. PMID- 9726973 TI - Stress-activated protein kinase/Jun N-terminal kinase is required for interleukin (IL)-1-induced IL-6 and IL-8 gene expression in the human epidermal carcinoma cell line KB. AB - The cytokine interleukin-1 (IL-1) is a major inflammatory hormone which activates a broad range of genes during inflammation. The signaling mechanisms triggered by IL-1 include activation of several distinct protein kinase systems. The stress activated protein kinase (SAPK), also termed Jun N-terminal kinase (JNK), is activated particularly strongly by the cytokine. In an attempt to delineate its role in activation of gene expression by IL-1, we inhibited the IL-1-induced SAPK/JNK activity by stable overexpression of either a catalytically inactive mutant of SAPKbeta (SAPKbeta(K-R)) or antisense RNA to SAPKbeta in human epidermal carcinoma cells. A detailed analysis of signal transduction in those cells showed that activation of neither NFkappaB nor p38 mitogen-activated protein kinase was affected, suggesting that we achieved specific blockade of the SAPK/JNK. In untransfected and vector-transfected KB cells, IL-1 induced a strong increase in expression of IL-6 and IL-8 mRNA, along with the synthesis of high amounts of the proteins. In two KB cell clones stably overexpressing the mutant SAPKbeta(K-R), and three clones stably overexpressing antisense RNA to SAPKbeta, expression of IL-6 and IL-8 in response to IL-1 was strongly reduced at both the mRNA and protein level. These data indicate that the SAPK/JNK pathway provides an indispensable signal for IL-1-induced expression of IL-6 and IL-8. PMID- 9726975 TI - Structural analysis of sequences O-linked to mannose reveals a novel Lewis X structure in cranin (dystroglycan) purified from sheep brain. AB - The Lewis X epitope, Galbeta1-4(Fucalpha1-3)GlcNAc-R, has been implicated in cell cell recognition events in a number of systems including the central nervous system and is expressed on diverse glycoconjugates including cell adhesion molecules, glycolipids, and the proteoglycan phosphacan. Although Lewis X sequences 3-linked to mannose have been described within proteoglycan fractions of mammalian brain, these have not been reported in other contexts and have been widely believed to be peculiar constituents of brain proteoglycans. In the present paper, we confirm the existence of Lewis X structures O-linked to mannose within the mammalian brain, demonstrate that these structures are present on a well defined mucin-like glycoprotein, cranin (dystroglycan), and report studies suggesting that the linkages involved may be predominantly 2-linked to mannose. Mannose-linked Lewis X is the latest in an increasing list of oligosaccharide recognition "tags" that have been shown to be expressed on cranin (dystroglycan) purified from brain. PMID- 9726976 TI - Bcl-xS and Bad potentiate the death suppressing activities of Bcl-xL, Bcl-2, and A1 in yeast. AB - Members of the Bcl-2 family can be grouped into three classes based upon their effects on cell death. The first class suppresses death and includes Bcl-2. A second group, which includes Bax, is lethal, whereas a third class, including Bcl xS, potentiates killing, although the members are not lethal by themselves. The proteins in the last class are proposed to exert their activity by binding to anti-apoptotic family members, thereby making the cell more susceptible to killing by another agent. To test this hypothesis, an inducible yeast expression system is reported that permits the functional analysis of three Bcl-2 family members. In yeast, Bax is lethal, and this activity is suppressed by Bcl-xL, Bcl 2, and A1. Co-expression of Bcl-xS did not diminish the ability of any of the anti-apoptotic members to antagonize Bax. Rather, co-expression of Bcl-xS potentiated the anti-death activity of all three proteins. This effect was not the result of changes in either the levels or integrity of Bax or anti-apoptotic proteins. Thus, Bcl-xS can bind to anti-apoptotic family members, but this association does not result in loss of biological activity. Therefore, Bcl-xS may act downstream of Bax and in a pathway that is conserved in yeast. PMID- 9726977 TI - Characterization of human TCR Vbeta gene promoter. Role of the dodecamer motif in promoter activity. AB - During T-lymphocyte development, the T-cell antigen receptor (TCR) gene expression is controlled by its promoter and enhancer elements and regulated in tissue- and development stage-specific manner. To uncover the promoter function and to define positive and negative regulatory elements in TCR gene promoters, the promoter activities from 13 human TCR Vbeta genes were determined by the transient transfection system and luciferase reporter assay. Although most of the TCR Vbeta gene promoters that we tested are inactive by themselves, some promoters were found to be constitutively strong. Among them, Vbeta6.7 is the strongest. 5'-Deletion and fragmentation experiments have narrowed the full promoter activity of Vbeta6.7 to a fragment of 147 base pairs immediately 5' to the transcription initiation site. A decanucleotide motif with the consensus sequence AGTGAYRTCA has been found to be conserved in most TCR Vbeta gene promoters. There are three such decamer motifs in the promoter region of Vbeta6.7, and the contribution of each such motif to the promoter activity has been examined. Further site-directed mutagenesis analyses showed that: 1) when two Ts in the decamer were mutated, the promoter activity was totally abolished; 2) when two additional nucleotides 3' to the end of decamer were mutated, the promoter activity was decreased to two-thirds of the full level; and 3) when the element with the sequence AGTGATGTCACT was inserted into other promoters, the original weak promoters become very strong. Taken together, our data suggest that the positive regulatory element in Vbeta6.7 should be considered a dodecamer rather than a decamer and that it confers strong basal transcriptional activity on TCR Vbeta genes. PMID- 9726978 TI - Metalloregulation of FRE1 and FRE2 homologs in Saccharomyces cerevisiae. AB - The high affinity uptake systems for iron and copper ions in Saccharomyces cerevisiae involve metal-specific permeases and two known cell surface Cu(II) and Fe(III) metalloreductases, Fre1 and Fre2. Five novel genes found in the S. cerevisiae genome exhibit marked sequence similarity to Fre1 and Fre2, suggesting that the homologs are part of a family of proteins related to Fre1 and Fre2. The homologs are expressed genes in S. cerevisiae, and their expression is metalloregulated as is true with FRE1 and FRE2. Four of the homologs (FRE3-FRE6) are specifically iron-regulated through the Aft1 transcription factor. These genes are expressed either in cells limited for iron ion uptake by treatment with a chelator or in cells lacking the high affinity iron uptake system. Expression of FRE3-FRE6 is elevated in AFT1-1 cells and attenuated in aft1 null cells, showing that iron modulation occurs through the Aft1 transcriptional activator. The fifth homolog FRE7 is specifically copper-metalloregulated. FRE7 is expressed in cells limited in copper ion uptake by a Cu(I)-specific chelator or in cells lacking the high affinity Cu(I) permeases. The constitutive expression of FRE7 in MAC1 cells and the lack of expression in mac1-1 cells are consistent with Mac1 being the critical transcriptional activator of FRE7 expression. The 5' promoter sequence of FRE7 contains three copper-responsive promoter elements. Two elements are critical for Mac1-dependent FRE7 expression. Combinations of either the distal and central elements or the central and proximal elements result in copper regulated FRE7 expression. Spacing between Mac1-responsive sites is important as shown by the attenuated expression of FRE7 and CTR1 when two elements are separated by over 100 base pairs. From the three Mac1-responsive elements in FRE7, a new consensus sequence for Mac1 binding can be established as TTTGC(T/G)C(A/G). PMID- 9726979 TI - Molecular cloning and functional characterization of murine sphingosine kinase. AB - Sphingosine-1-phosphate (SPP) is a novel lipid messenger that has dual function. Intracellularly it regulates proliferation and survival, and extracellularly, it is a ligand for the G protein-coupled receptor Edg-1. Based on peptide sequences obtained from purified rat kidney sphingosine kinase, the enzyme that regulates SPP levels, we report here the cloning, identification, and characterization of the first mammalian sphingosine kinases (murine SPHK1a and SPHK1b). Sequence analysis indicates that these are novel kinases, which are not similar to other known kinases, and that they are evolutionarily conserved. Comparison with Saccharomyces cerevisiae and Caenorhabditis elegans sphingosine kinase sequences shows that several blocks are highly conserved in all of these sequences. One of these blocks contains an invariant, positively charged motif, GGKGK, which may be part of the ATP binding site. From Northern blot analysis of multiple mouse tissues, we observed that expression was highest in adult lung and spleen, with barely detectable levels in skeletal muscle and liver. Human embryonic kidney cells and NIH 3T3 fibroblasts transiently transfected with either sphingosine kinase expression vectors had marked increases (more than 100-fold) in sphingosine kinase activity. The enzyme specifically phosphorylated D-erythro sphingosine and did not catalyze the phosphorylation of phosphatidylinositol, diacylglycerol, ceramide, D,L-threo-dihydrosphingosine or N, N dimethylsphingosine. The latter two sphingolipids were competitive inhibitors of sphingosine kinase in the transfected cells as was previously found with the purified rat kidney enzyme. Transfected cells also had a marked increase in mass levels of SPP with a concomitant decrease in levels of sphingosine and, to a lesser extent, in ceramide levels. Our data suggest that sphingosine kinase is a prototypical member of a new class of lipid kinases. Cloning of sphingosine kinase is an important step in corroborating the intracellular role of SPP as a second messenger. PMID- 9726980 TI - Nuleclear extracts of Crithidia fasciculata contain a factor(s) that binds to the 5'-untranslated regions of TOP2 and RPA1 mRNAs containing sequences required for their cell cycle regulation. AB - The Crithidia fasciculata replication protein A gene, RPA1, and topoisomerase II gene, TOP2, encode proteins involved in the replication of nuclear and mitochondrial DNA, respectively. Transcripts of both genes accumulate periodically during the cell cycle and attain their maximum levels just before S phase. Octamer consensus sequences within the 5'-untranslated region (UTR) of both genes have been shown to be necessary for cycling of these transcripts. Using a gel retardation assay, we show here that nuclear extracts of C. fasciculata contain a protein factor(s) that binds specifically to RNA from 5' UTRs of TOP2 and RPA1 genes. In addition, mutations in the consensus octamer sequence abolish binding to the RNA in both cases. Ultraviolet cross-linking using a radiolabeled TOP2 5'-UTR probe identified proteins with apparent molecular masses of 74 and 37 kDa in the RNA-protein complex. Nuclear extracts prepared from synchronized cells show that the binding activity varies during the cell cycle in parallel with TOP2 and RPA1 mRNA levels. These results suggest that the cell cycle regulation of the mRNA levels of trypanosomatid DNA replication genes may be mediated by binding of specific proteins to conserved sequences in the 5'-UTR of their transcripts. PMID- 9726981 TI - Tyrosine structural changes detected during the photoactivation of rhodopsin. AB - We present the first Fourier transform infrared (FTIR) analysis of an isotope labeled eukaryotic membrane protein. A combination of isotope labeling and FTIR difference spectroscopy was used to investigate the possible involvement of tyrosines in the photoactivation of rhodopsin (Rho). Rho --> MII difference spectra were obtained at 10 degrees C for unlabeled recombinant Rho and isotope labeled L-[ring-2H4]Tyr-Rho expressed in Spodoptera frugiperda cells grown on a stringent culture medium containing enriched L-[ring-2H4]Tyr and isolated using a His6 tag. A comparison of these difference spectra revealed reproducible changes in bands that correspond to tyrosine and tyrosinate vibrational modes. A similar pattern of tyrosine/tyrosinate bands has also been observed in the bR --> M transition in bacteriorhodopsin, although the sign of the bands is reversed. In bacteriorhodopsin, these bands were assigned to Tyr-185, which along with Pro-186 in the F-helix, may form a hinge that facilitates alpha-helix movement. PMID- 9726982 TI - Properties of human hemoglobins with increased polarity in the alpha- or beta heme pocket. Carbonmonoxy derivatives. AB - The spectroscopic, conformational, and functional properties of mutant carbonmonoxy hemoglobins in which either the beta-globin Val67(E11) or the alpha globin Val62(E11) is replaced by threonine have been investigated. The thermal evolution of the Soret absorption band and the stretching frequency of the bound CO were used to probe the stereodynamic properties of the heme pocket. The functional properties were investigated by kinetic measurements. The spectroscopic and functional data were related to the conformational properties through molecular analysis. The effects of this nonpolar-to-polar isosteric mutation are: (i) increase of heme pocket anharmonic motions, (ii) stabilization of the A0 conformer in the IR spectrum, (iii) increased CO dissociation rates. The spectroscopic data indicate that for the carbonmonoxy derivatives, the Val - > Thr mutation has a larger conformational effect on the beta-subunits than on the alpha-subunits. This is at variance with the deoxy derivatives where the conformational modification was larger in the heme pocket of the alpha-subunit (Cupane, A., Leone, M., Militello, V., Friedman, R. K., Koley, A. P., Vasquez, G. P., Brinigar, W. S., Karavitis, M., and Fronticelli, C. (1997) J. Biol. Chem. 272, 26271-26278). These effects are attributed to a different electrostatic interaction between Ogamma of Thr(E11) and the bound CO molecule. Molecular analysis indicates a more favorable interaction of the bound CO with Thr Ogamma in the beta-subunit heme pocket. PMID- 9726983 TI - Phosphoinositide 3-kinase regulates phospholipase Cgamma-mediated calcium signaling. AB - It has been demonstrated that the lipid products of the phosphoinositide 3-kinase (PI3K) can associate with the Src homology 2 (SH2) domains of specific signaling molecules and modify their actions. In the current experiments, phosphatidylinositol 3,4, 5-trisphosphate (PtdIns-3,4,5-P3) was found to bind to the C-terminal SH2 domain of phospholipase Cgamma (PLCgamma) with an apparent Kd of 2.4 microM and to displace the C-terminal SH2 domain from the activated platelet-derived growth factor receptor (PDGFR). To investigate the in vivo relevance of this observation, intracellular inositol trisphosphate (IP3) generation and calcium release were examined in HepG2 cells expressing a series of PDGFR mutants that activate PLCgamma with or without receptor association with PI3K. Coactivation of PLCgamma and PI3K resulted in an approximately 40% increase in both intracellular IP3 generation and intracellular calcium release as compared with selective activation of PLCgamma. Similarly, the addition of wortmannin or LY294002 to cells expressing the wild-type PDGFR inhibited the release of intracellular calcium. Thus, generation of PtdIns-3,4,5-P3 by receptor associated PI3K causes an increase in IP3 production and intracellular calcium release, potentially via enhanced PtdIns-4, 5-P2 substrate availability due to PtdIns-3,4,5-P3-mediated recruitment of PLCgamma to the lipid bilayer. PMID- 9726985 TI - Inhibition of prothrombinase by human secretory phospholipase A2 involves binding to factor Xa. AB - Human group II secretory phospholipase A2 (hsPLA2) exhibits significant anticoagulant activity that does not require its enzymatic activity. We examined which coagulation factor was targeted by hsPLA2 and analyzed which region of the protein may be involved in this inhibition. Prothrombin time coagulation assays indicated that hsPLA2 did not inhibit activated factor V (FVa) activity, whereas activated factor X (FXa) one-stage coagulation assays suggested that FXa was inhibited. The inhibitory effect of hsPLA2 on prothrombinase activity of FXa, FV, phospholipids, and Ca2+ complex was markedly enhanced upon preincubation of hsPLA2 with FXa but not with FV. Prothrombinase activity was also strongly inhibited by hsPLA2 in the absence of PL. High concentrations of FVa in the prothrombinase generation assay reversed the inhibitory effect of hsPLA2. By using isothermal titration calorimetry, we demonstrated that hsPLA2 binds to FXa in solution with a 1:1 stoichiometry and a Kd of 230 nM. By using surface plasmon resonance we determined the rate constants, kon and koff, of the FXa/hsPLA2 interaction and analyzed the Ca2+ effect on these constants. When preincubated with FXa, synthetic peptides comprising residues 51-74 and 51-62 of hsPLA2 inhibited prothrombinase assays, providing evidence that this part of the molecule, which shares similarities with a region of FVa that binds to FXa, is likely involved in the anticoagulant interaction of hsPLA2 with FXa. In conclusion, we propose that residues 51-62 of hsPLA2 bind to FXa at a FVa-binding site and that hsPLA2 decreases the prothrombinase generation by preventing FXa.FVa complex formation. PMID- 9726984 TI - Inhibition of hepatocytic autophagy by adenosine, aminoimidazole-4-carboxamide riboside, and N6-mercaptopurine riboside. Evidence for involvement of amp activated protein kinase. AB - To examine the role of AMP-activated protein kinase (AMPK; EC 2.7.1. 109) in the regulation of autophagy, rat hepatocytes were incubated with the AMPK proactivators, adenosine, 5-amino-4-imidazole carboxamide riboside (AICAR), or N6 mercaptopurine riboside. Autophagic activity was inhibited by all three nucleosides, AICAR and N6-mercaptopurine riboside being more potent (IC50 = 0.3 mM) than adenosine (IC50 = 1 mM). 2'-Deoxycoformycin, an adenosine deaminase (EC 3.5.4.4) inhibitor, increased the potency of adenosine 5-fold, suggesting that the effectiveness of adenosine as an autophagy inhibitor was curtailed by its intracellular deamination. 5-Iodotubercidin, an adenosine kinase (EC 2.7.1.20) inhibitor, abolished the effects of all three nucleosides, indicating that they needed to be phosphorylated to inhibit autophagy. A 5-iodotubercidin-suppressible phosphorylation of AICAR to 5-aminoimidazole-4-carboxamide riboside monophosphate was confirmed by chromatographic analysis. AICAR, up to 0.4 mM, had no significant effect on intracellular ATP concentrations. Because activated AMPK phosphorylates and inactivates 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase (EC 1.1.1.88), the rate-limiting enzyme in cholesterol synthesis, the strong inhibition of hepatocytic cholesterol synthesis by all three nucleosides confirmed their ability to activate AMPK under the conditions used. Lovastatin and simvastatin, inhibitors of HMG-CoA reductase, strongly suppressed cholesterol synthesis while having no effect on autophagic activity, suggesting that AMPK inhibits autophagy independently of its effects on HMG-CoA reductase and cholesterol metabolism. PMID- 9726986 TI - Binding site for S-adenosyl-L-methionine in a central region of mammalian reovirus lambda2 protein. Evidence for activities in mRNA cap methylation. AB - One or more proteins in mammalian reovirus core particles mediate two RNA methylation activities, (guanosine-7-N)-methyltransferase and (guanosine-2'-O) methyltransferase, that contribute to forming the 5' cap 1 structure on viral mRNA. We used UV irradiation to identify core proteins that bind S-adenosyl-L methionine (SAM), the methyl-group donor for both methyltransferases. A [methyl 3H]SAM-binding site was observed among the reovirus lambda proteins; was shown to be specific by competition with low levels of S-adenosyl-L-homocysteine, the product of methyl-group transfer from SAM; and was subsequently localized to protein lambda2. lambda2 mediates the guanylyltransferase reaction in cap formation and was previously proposed to mediate one or both methylation reactions as well. SAM binding was demonstrated for both lambda2 in cores and lambda2 expressed in insect cells from a recombinant baculovirus. Using three different methods to cleave lambda2, a binding site for SAM was tentatively localized to a central region of lambda2, between residues 792 and 1100, which includes a smaller region with sequence similarity to the SAM-binding pocket of other methyltransferases. Alanine substitutions at positions 827 and 829 within this predicted binding region greatly reduced the capacity of baculovirus expressed lambda2 protein to undergo UV cross-linking to SAM but had no effects on either the guanylyltransferase activity of this protein or its conformation as judged by partial proteolysis, suggesting that one or both of these residues is essential for SAM binding. Based on these findings, we propose that the two methyltransferase activities involved in mRNA capping by reovirus cores utilize a single SAM-binding pocket within a central region of lambda2. PMID- 9726987 TI - A human SPT3-TAFII31-GCN5-L acetylase complex distinct from transcription factor IID. AB - In yeast, SPT3 is a component of the multiprotein SPT-ADA-GCN5 acetyltransferase (SAGA) complex that integrates proteins with transcription coactivator/adaptor functions (ADAs and GCN5), histone acetyltransferase activity (GCN5), and core promoter-selective functions (SPTs) involving interactions with the TATA-binding protein (TBP). In particular, yeast SPT3 has been shown to interact directly with TBP. Here we report the molecular cloning of a cDNA encoding a human homologue of yeast SPT3. Amino acid sequence comparisons between human SPT3 (hSPT3) and its counterparts in different yeast species reveal three highly conserved domains, with the most conserved 92-amino acid N-terminal domain being 25% identical with human TAFII18. Despite the significant sequence similarity with TAFII18, native hSPT3 is not a bona fide TAFII because it is not associated in vivo either with human TBP/TFIID or with a TFIID-related TBP-free TAFII complex. However, we present evidence that hSPT3 is associated in vivo with TAFII31 and the recently described longer form of human GCN5 (hGCN5-L) in a novel human complex that has histone acetyltransferase activity. We propose that the human SPT3-TAFII31-GCN5-L acetyltransferase (STAGA) complex is a likely homologue of the yeast SAGA complex. PMID- 9726988 TI - Fatty acids activate transcription of the muscle carnitine palmitoyltransferase I gene in cardiac myocytes via the peroxisome proliferator-activated receptor alpha. AB - To explore the gene regulatory mechanisms involved in the metabolic control of cardiac fatty acid oxidative flux, the expression of muscle-type carnitine palmitoyltransferase I (M-CPT I) was characterized in primary cardiac myocytes in culture following exposure to the long-chain mono-unsaturated fatty acid, oleate. Oleate induced steady-state levels of M-CPT I mRNA 4.5-fold. The transcription of a plasmid construct containing the human M-CPT I gene promoter region fused to a luciferase gene reporter transfected into cardiac myocytes, was induced over 20 fold by long-chain fatty acid in a concentration-dependent and fatty acyl-chain length-specific manner. The M-CPT I gene promoter fatty acid response element (FARE-1) was localized to a hexameric repeat sequence located between 775 and 763 base pairs upstream of the initiator codon. Cotransfection experiments with expression vectors for the peroxisome proliferator-activated receptor alpha (PPARalpha) demonstrated that FARE-1 is a PPARalpha response element capable of conferring oleate-mediated transcriptional activation to homologous or heterologous promoters. Electrophoretic mobility shift assays demonstrated that PPARalpha bound FARE-1 with the retinoid X receptor alpha. The expression of M CPT I in hearts of mice null for PPARalpha was approximately 50% lower than levels in wild-type controls. Moreover, a PPARalpha activator did not induce cardiac expression of the M-CPT I gene in the PPARalpha null mice. These results demonstrate that long-chain fatty acids regulate the transcription of a gene encoding a pivotal enzyme in the mitochondrial fatty acid uptake pathway in cardiac myocytes and define a role for PPARalpha in the control of myocardial lipid metabolism. PMID- 9726989 TI - Transcription factor erythroid Kruppel-like factor (EKLF) is essential for the erythropoietin-induced hemoglobin production but not for proliferation, viability, or morphological maturation. AB - The erythroid Kruppel-like factor (EKLF) is essential for the transcription of betamaj globin in erythroid cells. We show here that RNA for this transcription factor did not alter during erythropoietin-induced differentiation of J2E cells; however, EKLF protein content decreased and was inversely related to globin production. This unexpected result was also observed during chemically induced maturation of two murine erythroleukemia cell lines. To explore the role of EKLF in erythroid terminal differentiation, an antisense EKLF construct was introduced into J2E cells. As a consequence EKLF RNA and protein levels fell by approximately 80%, and the cells were unable to manufacture hemoglobin in response to erythropoietin. The failure to produce hemoglobin was due to reduced transcription of not only globin genes but also key heme enzyme genes. However, numerous other genes, including several erythroid transcription factors, were unaffected by the decrease in EKLF. Although hemoglobin synthesis was severely impaired with depleted EKLF levels, morphological maturation in response to erythropoietin continued normally. Moreover, erythropoietin-induced proliferation and viability were unaffected by the decrease in EKLF levels. We conclude that EKLF affects a specific set of genes, which regulates hemoglobin production and has no obvious effect on morphological changes, cell division, or viability in response to erythropoietin. PMID- 9726990 TI - Identification of CX3CR1. A chemotactic receptor for the human CX3C chemokine fractalkine and a fusion coreceptor for HIV-1. AB - Fractalkine is a multimodular human leukocyte chemoattractant protein and a member of the chemokine superfamily. Unlike other human chemokines, the chemokine domain of fractalkine has three amino acids between two conserved cysteines, referred to as the CX3C motif. Both plasma membrane-associated and shed forms of fractalkine have been identified. Here, we show that the recombinant 76-amino acid chemokine domain of fractalkine is a potent and highly specific chemotactic agonist at a human orphan receptor previously named V28 or alternatively CMKBRL1 (chemokine beta receptor-like 1), which was shown previously to be expressed in neutrophils, monocytes, T lymphocytes, and several solid organs, including brain. CMKBRL1/V28 also functioned with CD4 as a coreceptor for the envelope protein from a primary isolate of HIV-1 in a cell-cell fusion assay, and fusion was potently and specifically inhibited by fractalkine. Thus CMKBRL1/V28 is a specific receptor for fractalkine, and we propose to rename it CX3CR1 (CX3C chemokine receptor 1), according to an accepted nomenclature system. PMID- 9726991 TI - Mapping of the DNA binding domain of the copper-responsive transcription factor Mac1 from Saccharomyces cerevisiae. AB - Mac1 from Saccharomyces cerevisiae activates transcription of genes, including CTR1 in copper-deficient cells. N-terminal fusions of Mac1 with the herpes simplex VP16 activation domain were used to show that residues 1-159 in Mac1 constitute the minimal DNA binding domain. Mac1-(1-159) purified from Escherichia coli contains two bound Zn(II) ions. Electrophoretic mobility shift assays showed direct and specific binding by Mac1-(1-159) to a DNA duplex containing the copper responsive element TTTGCTCA. The DNA binding affinity of Mac1-(1-159) for a duplex containing a single promoter element or an inverted repeat was 5 nM for the 1:1 complex. The N-terminal 40-residue segment of Mac1 is homologous to the DNA binding zinc module found in the copper-activated transcription factors Ace1 and Amt1. A MAC1 mutation yielding a Cys11 --> Tyr substitution at the first candidate zinc ligand position relative to Ace1 resulted in a loss of in vivo function. Two TTTGCTCA promoter elements are necessary for efficient Mac1 mediated transcriptional activation. The elements appear to function synergistically. Increasing the number of elements yields more than additive enhancements in CTR1 expression. PMID- 9726992 TI - The soluble alpha-glycerophosphate oxidase from Enterococcus casseliflavus. Sequence homology with the membrane-associated dehydrogenase and kinetic analysis of the recombinant enzyme. AB - The soluble flavoprotein alpha-glycerophosphate oxidase from Enterococcus casseliflavus catalyzes the oxidation of a "non-activated" secondary alcohol, in contrast to the flavin-dependent alpha-hydroxy- and alpha-amino acid oxidases. Surprisingly, the alpha-glycerophosphate oxidase sequence is 43% identical to that of the membrane-associated alpha-glycerophosphate dehydrogenase from Bacillus subtilis; only low levels of identity (17-22%) result from comparisons with other FAD-dependent oxidases. The recombinant alpha-glycerophosphate oxidase is fully active and stabilizes a flavin N(5)-sulfite adduct, but only small amounts of intermediate flavin semiquinone are observed during reductive titrations. Direct determination of the redox potential for the FAD/FADH2 couple yields a value of -118 mV; the protein environment raises the flavin potential by 100 mV in order to provide for a productive interaction with the reducing substrate. Steady-state kinetic analysis, using the enzyme-monitored turnover method, indicates that a ping-pong mechanism applies and also allows the determination of the corresponding kinetic constants. In addition, stopped-flow studies of the reductive half-reaction provide for the measurement of the dissociation constant for the enzyme. alpha-glycerophosphate complex and the rate constant for reduction of the enzyme flavin. These and other results demonstrate that this enzyme offers a very promising paradigm for examining the protein determinants for flavin reactivity and mechanism in the energy-yielding metabolism of alpha-glycerophosphate. PMID- 9726993 TI - Interaction of the clathrin-coated vesicle V-ATPase with ADP and sodium azide. AB - The kinetics of adenosine triphosphate (ATP)-dependent proton transport into clathrin-coated vesicles from bovine brain have been studied. We observe that the vacuolar proton-translocating ATPase (V-ATPase) from clathrin-coated vesicles is subject to two different types of inhibition by ADP. The first is competitive inhibition with respect to ATP, with a Ki for ADP of 11 microM. The second type of inhibition occurs after preincubation of the V-ATPase in the presence of ADP and Mg2+, which results in inhibition of the initial rate of proton transport followed by reactivation over the course of several minutes. The second effect is observed at ADP concentrations as low as 0.1-0.2 microM, indicating that a high affinity inhibitory complex is formed between ADP and the V-ATPase and is only slowly dissociated after the addition of ATP. We have further investigated the effect of sodium azide, an inhibitor of the F-ATPases that has been shown to stabilize an inactive complex between ADP and the F1-F0-ATP synthase (F-ATPase). We observed that azide inhibited ATP-dependent proton transport by the purified, reconstituted V-ATPase with a K0.5 of 0.2-0.4 mM but had no effect on ATP hydrolysis. Azide was shown not to increase the passive proton permeability of reconstituted vesicles and did not stimulate ATP hydrolysis by the reconstituted enzyme, in contrast with CCCP, which both abolished the proton gradient and stimulated hydrolysis. Thus, azide does not appear to act as a simple uncoupler of proton transport and ATP hydrolysis. Rather, azide may have some more direct effect on V-ATPase activity. Possible mechanisms by which azide could exert this effect on the V-ATPase and the contrasting effects of azide on the F- and V ATPases are discussed. PMID- 9726994 TI - Differential cross-regulation of the human chemokine receptors CXCR1 and CXCR2. Evidence for time-dependent signal generation. AB - Neutrophils and transfected RBL-2H3 cells were used to investigate the mechanism of cross-regulation of the human interleukin-8 (IL-8) receptors CXCR1 and CXCR2 by chemoattractants. In neutrophils, Ca2+ mobilization by the CXCR2-specific chemokine, growth-related oncogene alpha (Groalpha), was desensitized by prior exposure to the chemoattractants N-formylated peptides (fMLP) or a complement cleavage product (C5a). In contrast, growth-related oncogene alpha did not desensitize the latter receptors. To investigate this phenomenon, CXCR2 was stably expressed in RBL-2H3 cells and mediated phosphoinositide hydrolysis, Ca2+ mobilization, chemotaxis, and secretion. In cells co-expressing CXCR2 and receptors for either C5a (C5aR) or fMLP (FR), CXCR2 was cross-phosphorylated and cross-desensitized by C5a and fMLP. However, neither C5aR nor FR was cross phosphorylated or cross-desensitized by CXCR2 activation, although CXCR1 did mediate this process. Receptor internalization induced by IL-8 was more rapid and occurred at lower doses with CXCR2 than CXCR1, although both receptors mediated equipotent chemotaxis and exocytosis in RBL. Truncation of the cytoplasmic tail of CXCR2 (331T) prolonged its signaling relative to CXCR2, increased its resistance to internalization, and induced phospholipase D activation. 331T was resistant to homologous phosphorylation and cross-phosphorylation but not cross desensitization of its Ca2+ mobilization by fMLP or C5a, indicating an inhibitory site distal to receptor/G protein coupling. In contrast to CXCR2, stimulation of 331T cross-desensitized Ca2+ mobilization by both FR and C5aR. CXCR2 and the mutant 331T induced phospholipase C beta3 phosphorylation to an extent equivalent to that of CXCR1. Taken together, these results suggest that CXCR1 and CXCR2 bind IL-8 to produce a group of equipotent responses, but their ability to generate other signals, including receptor internalization, cross-desensitization, and phospholipase D activation, are very different. The latter phenomena apparently require prolonged receptor activation, which in the case of CXCR2 is precluded by rapid receptor phosphorylation and internalization. Thus, receptors coupling to identical G proteins may trigger different cellular responses dependent on the length of their signaling time, which can be regulated by receptor phosphorylation. PMID- 9726995 TI - Hypoxia-inducible mammalian gene expression analyzed in vivo at a TATA-driven promoter and at an initiator-driven promoter. AB - We have analyzed protein-DNA interactions in vivo at transcriptional control elements for two hypoxia-inducible genes in mouse hepatoma cells. The promoter for the phosphoglycerate kinase 1 (PGK1) gene contains an initiator element, but no TATA sequence, whereas the promoter for the glucose transporter 1 (Glut1) gene contains a TATA element but no initiator sequence. Our findings reveal hypoxia inducible, Arnt-dependent occupancy of DNA recognition sites for hypoxia inducible factor 1 (HIF-1) upstream of both target genes. The conserved recognition motif among the five recognition sites is 5'-CGTG-3'. The PGK1 promoter exhibits constitutive occupancy of a binding site for an unknown protein(s); however, we detect no protein-DNA interaction at the initiator element, in either uninduced or induced cells. The Glut1 promoter also exhibits constitutive protein binding; in addition, the TATA element exhibits partial occupancy in uninduced cells and increased occupancy under hypoxic conditions. We find no evidence for hypoxia-induced changes in chromatin structure of either gene. Time-course analyses of the Glut1 gene reveal a temporal relationship between occupancy of HIF-1 sites and TATA element occupancy. Our findings suggest that the promoters for both hypoxia-responsive genes constitutively maintain an accessible chromatin configuration and that HIF-1 facilitates transcription by recruiting and/or stabilizing a transcription factor(s), such as TFIID, at both promoters. PMID- 9726996 TI - Stimulation of ATPase activity of purified multidrug resistance-associated protein by nucleoside diphosphates. AB - Membrane vesicles prepared from cells expressing the multidrug resistance associated protein (MRP) transport glutathione S-conjugates of hydrophobic substrates in an ATP dependent manner. Purified MRP possesses ATPase activity which can be further stimulated by anticancer drugs or leukotriene C4. However, the detailed relationship between ATP hydrolysis and drug transport has not been established. How the ATPase activity of MRP is regulated in the cell is also not known. In this report, we have examined the effects of different nucleotides on the ATPase activity of purified MRP. We have found that pyrimidine nucleoside triphosphates have little effect on enzymatic activity. In contrast, purine nucleotides dATP, dGTP, and adenosine 5'-(beta,gamma-imido)triphosphate function as competitive inhibitors. Somewhat unexpectedly, low concentrations of all the nucleoside diphosphates (NDPs) tested, except UDP, stimulate the ATPase activity severalfold. ADP or GDP at higher concentrations was inhibitory, reflecting NDP binding to the substrate site. On the other hand, the enhancement of hydrolysis at low NDP concentrations must reflect interactions with a separate site. Therefore, we postulate the presence of at least two types of nucleotide binding sites on the MRP, a catalytic site(s) to which ATP preferentially binds and is hydrolyzed and a regulatory site to which NDPs preferentially bind and stimulate hydrolysis. Interestingly, the stimulatory effects of drugs transported by MRP and NDPs are not additive, i.e. drugs are not able to further stimulate the NDP activated enzyme. Hence, the two activation pathways intersect at some point. Since both nucleotide binding domains of MRP are likely to be required for drug stimulation of ATPase activity, the two sites that we postulate may also involve both domains. PMID- 9726997 TI - Phosphorylation of the Spo0B response regulator phosphotransferase of the phosphorelay initiating development in Bacillus subtilis. AB - The initiation of sporulation in Bacillus subtilis is regulated by the phosphorelay, a complex signal transduction system consisting of kinases and response regulators. The key component of a phosphorelay is the phosphotransferase, which recognizes two response regulators and transfers a phosphoryl group between them. In this reaction, the phosphoryl of one response regulator is transferred to a histidine on the phosphotransferase before phosphorylating an aspartate of the second response regulator. The phosphorylated histidine on the Spo0B phosphotransferase was found to be His30. Site-directed mutation of His30 to alanine destroyed its phosphotransferase activity in vitro and strains constructed with this mutation were unable to sporulate. None of the other 10 histidines of Spo0B was implicated in the phosphotransferase reaction. A structurally vulnerable site, histidine 23, was also identified through the mutational study. The His30 of Spo0B resides in a domain with little sequence homology to functionally equivalent domains in the phosphorelays of other bacterial and yeast systems, suggesting that the two types of phosphotransfer domains evolved convergently. PMID- 9726998 TI - Apolipoprotein(a) binds via its C-terminal domain to the protein core of the proteoglycan decorin. Implications for the retention of lipoprotein(a) in atherosclerotic lesions. AB - Although it is known that lipoprotein(a) (Lp(a)) binds to proteoglycans, the mechanism for this binding has not been fully elucidated. In order to shed light on this subject, we examined the interactions of decorin, a proteoglycan with a well defined protein core and a single glycosaminoglycan (GAG) chain, with Lp(a) and derivatives, namely Lp(a) deprived of apo(a), or Lp(a-), free apo(a), and the two main proteolytic fragments, F1 and F2. By circular dichroism criteria, the decorin preparations used had the same secondary structure as that previously reported for native decorin. Authentic low density lipoprotein from the same human donor was used as a control. In a solid phase system, Lp(a-)and low density lipoprotein bound to decorin in a comparable manner. This binding required Ca2+/Mg2+ ions, was lysine-mediated, and was markedly decreased in the presence of GAG-depleted decorin, suggesting the ionic nature of the interaction likely involving apoB100 and the GAG component of decorin. Free apo(a) also bound to decorin; however, the binding was neither cation-dependent nor lysine-mediated, unaffected by sialic acid depletion of apo(a), and markedly decreased when either reduced and alkylated apo(a) or reduced and alkylated decorin was used in the assay. Of note, the binding of apo(a) was unaffected when it was incubated with a spectrally native decorin that had been renatured from either 4 M guanidine hydrochloride by extensive dialysis or cooled from 65 to 25 degrees C. On the other hand, the binding significantly increased when decorin was depleted of GAGs, which by themselves had no affinity for apo(a). The binding of apo(a) to the decorin protein core was also elicited by the C-terminal domain of apo(a), and it was favored by high NaCl concentrations, 1 to 2 M. No binding was exhibited by the N-terminal domain accounting for the lack of effect of apo(a) size polymorphism on the binding. In the case of whole Lp(a), the binding to immobilized decorin was mostly GAG-dependent and ionic in nature. A minor contribution by apo(a) was detected when GAG-depleted decorin was used in the assay. Our results indicate that the binding of Lp(a) to decorin involves interactions both electrostatic (apoB100-GAG) and hydrophobic (apo(a)-decorin protein core), and that the binding of apo(a) requires decorin protein core to be in its native state. PMID- 9726999 TI - Probing the role of cysteine residues in the EcoP15I DNA methyltransferase. AB - Chemical modification using thiol-directed agents and site-directed mutagenesis has been used to investigate the role of cysteine residues of EcoP15I DNA methyltransferase. Irreversible inhibition of enzymatic activity was provoked by chemical modification of the enzyme by N-ethylmaleimide and iodoacetamide. 5, 5' Dithiobis(2-nitrobenzoic acid) titration of the enzyme under nondenaturing and denaturing conditions confirmed the presence of six cysteine residues without any disulfides in the protein. Aware that relatively bulky reagents inactivate the methyltransferase by directly occluding the substrate-binding site or by locking the methyltransferase in an inactive conformation, we used site-directed mutagenesis to sequentially replace each of the six cysteines in the protein at positions 30, 213, 344, 434, 553, and 577. All the resultant mutant methylases except for the C344S and C344A enzymes retained significant activity as assessed by in vivo and in vitro assays. The effects of the substitutions on the function of EcoP15I DNA methyltransferase were investigated by substrate binding assays, activity measurements, and steady-state kinetic analysis of catalysis. Our results clearly indicate that the cysteines at positions other than 344 are not essential for activity. In contrast, the C344A enzyme showed a marked loss of enzymatic activity. More importantly, whereas the inactive C344A mutant enzyme bound S-adenosyl-L-methionine, it failed to bind to DNA. Furthermore, in double and triple mutants where two or three cysteine residues were replaced by serine, all such mutants in which the cysteine at position 344 was changed, were inactive. Taken together, these results convincingly demonstrate that the Cys-344 is necessary for enzyme activity and indicate an essential role for it in DNA binding. PMID- 9727000 TI - Two-site interaction of nuclear factor of activated T cells with activated calcineurin. AB - Transcription factors belonging to the nuclear factor of activated T cells (NFAT) family regulate the expression of cytokine genes and other inducible genes during the immune response. The functions of NFAT proteins are directly controlled by the calcium- and calmodulin-dependent phosphatase calcineurin. Here we show that the binding of calcineurin to NFAT is substantially increased when calcineurin is activated with calmodulin and calcium. FK506.FKBP12 drug-immunophilin complexes inhibited the interaction of NFAT with activated calcineurin much more effectively than they inhibited the interaction with inactive calcineurin, suggesting that part of the interaction with activated calcineurin involved the enzyme active site. We have previously shown that NFAT is targeted to inactive calcineurin at a region distinct from the calcineurin active site (Aramburu, J., Garcia-Cozar, F. J., Raghavan, A., Okamura, H., Rao, A., and Hogan, P. G. (1998) Mol. Cell 1, 627-637); this region is also involved in NFAT binding to activated calcineurin, since binding is inhibited by an NFAT peptide spanning the calcineurin docking site on NFAT. The interacting surfaces are located on the catalytic domain of the calcineurin A chain and on an 86-amino acid fragment of the NFAT regulatory domain. NFAT binding to the calcineurin catalytic domain was inhibited by the calcineurin autoinhibitory domain and the RII substrate peptide, which bind in the calcineurin active site, as well as by the NFAT docking site peptide, which binds to a region of calcineurin distinct from the active site. We propose that, in resting cells, NFAT is targeted to a region of the calcineurin catalytic domain that does not overlap the calcineurin active site. Upon cell activation, displacement of the autoinhibitory domain by calmodulin binding allows NFAT to bind additionally to the calcineurin active site, thus positioning NFAT for immediate dephosphorylation at functional phosphoserine residues. PMID- 9727001 TI - A hyperprostaglandin E syndrome mutation in Kir1.1 (renal outer medullary potassium) channels reveals a crucial residue for channel function in Kir1.3 channels. AB - Loss of function mutations in kidney Kir1.1 (renal outer medullary potassium channel, KCNJ1) inwardly rectifying potassium channels can be found in patients suffering from hyperprostaglandin E syndrome (HPS), the antenatal form of Bartter syndrome. A novel mutation found in a sporadic case substitutes an asparagine by a positively charged lysine residue at amino acid position 124 in the extracellular M1-H5 linker region. When heterologously expressed in Xenopus oocytes and mammalian cells, current amplitudes from mutant Kir1.1a[N124K] channels were reduced by a factor of approximately 12 as compared with wild type. A lysine at the equivalent position is present in only one of the known Kir subunits, the newly identified Kir1.3, which is also poorly expressed in the recombinant system. When the lysine residue in guinea pig Kir1.3 (gpKir1.3) isolated from a genomic library was changed to an asparagine (reverse HPS mutation), mutant channels yielded macroscopic currents with amplitudes increased 6-fold. From single channel analysis it became apparent that the decrease in mutant Kir1.1 channels and the increase in mutant gpKir1.3 macroscopic currents were mainly due to the number of expressed functional channels. Coexpression experiments revealed a dominant-negative effect of Kir1.1a[N124K] and gpKir1.3 on macroscopic current amplitudes when coexpressed with wild type Kir1.1a and gpKir[K110N], respectively. Thus we postulate that in Kir1.3 channels the extracellular positively charged lysine is of crucial functional importance. The HPS phenotype in man can be explained by the lower expression of functional channels by the Kir1. 1a[N124K] mutant. PMID- 9727002 TI - Effect of insulin on farnesyltransferase. Specificity of insulin action and potentiation of nuclear effects of insulin-like growth factor-1, epidermal growth factor, and platelet-derived growth factor. AB - We have previously demonstrated that insulin activates farnesyltransferase (FTase) and augments the amounts of farnesylated p21 (Goalstone, M. L., and Draznin, B. (1996) J. Biol. Chem. 271, 27585-27589). We postulated that this aspect of insulin action might explain the "priming effect" of insulin on the cellular response to other growth factors. In the present study, we show the specificity of the effect of insulin on FTase. Insulin, but not insulin-like growth factor-1 (IGF-1), epidermal growth factor (EGF), or platelet-derived growth factor (PDGF), stimulated the phosphorylation of the alpha-subunit of FTase and the amounts of farnesylated p21. Even though all four growth factors utilized the Ras pathway to stimulate DNA synthesis, only insulin used this pathway to influence FTase. Insulin failed to stimulate FTase in cells expressing the chimeric insulin/IGF-1 receptor and in cells derived from the insulin receptor knock-out animals. Insulin potentiated the effects of IGF-1, EGF, and PDGF on DNA synthesis in cells expressing the wild type insulin receptor, but this potentiation was inhibited in the presence of the FTase inhibitor, alpha hydroxyfarnesylphosphonic acid. We conclude that the effect of insulin on FTase is specific, requires the presence of an intact insulin receptor, and serves as a conduit for the "priming" influence of insulin on the nuclear effects of other growth factors. PMID- 9727003 TI - Inhibition of 3T3-L1 adipocyte differentiation by expression of acyl-CoA-binding protein antisense RNA. AB - Several lines of evidence have recently underscored the significance of fatty acids or fatty acid-derived metabolites as signaling molecules in adipocyte differentiation. The acyl-CoA-binding protein (ACBP), which functions as an intracellular acyl-CoA pool former and transporter, is induced during adipocyte differentiation. In this report we describe the effects of expression of high levels of ACBP antisense RNA on the differentiation of 3T3-L1 cells. Pools of 3T3 L1 cells transfected with vectors expressing ACBP antisense RNA showed significantly less lipid accumulation as compared with cells transfected with the control vector. When individual clones were analyzed the degree of differentiation at day 10 was inversely correlated with the level of ACBP antisense RNA expression at day 0. Furthermore, in the clones with the highest levels of ACBP antisense expression, the induction of expression of the adipogenic transcription factors peroxisome proliferator-activated receptor gamma and CCAAT/enhancer-binding protein alpha as well as several adipocyte-specific genes was significantly delayed and reduced. The adipogenic potential of antisense-expressing cells was partially restored by transfection with a vector expressing high levels of ACBP. Taken together, these results are strong evidence that inhibition of differentiation is causally related to the decreased expression of ACBP, indicating that ACBP plays an important role during adipocyte differentiation. PMID- 9727004 TI - Mouse keratin 4 is necessary for internal epithelial integrity. AB - Keratins are intermediate filaments of epithelial cells. Mutations in keratin genes expressed in skin lead to human disorders, including epidermolysis bullosa simplex and epidermolytic hyperkeratosis. We examined the role of keratin 4 (K4) in maintaining the integrity of internal epithelial linings by using gene targeting to generate mice containing a null mutation in the epithelial K4 gene. Homozygous mice that do not express K4 develop a spectrum of phenotypes that affect several organs which express K4 including the esophagus, tongue, and cornea. The cellular phenotypes include basal hyperplasia, lack of maturation, hyperkeratosis, atypical nuclei, perinuclear clearing, and cell degeneration. These results are consistent with the notion that K4 is required for internal epithelial cell integrity. As mutations in K4 in humans lead to a disorder called white sponge nevus, the K4-deficient mice may serve as models for white sponge nevus and for understanding the role of K4 in cellular proliferation and differentiation. PMID- 9727005 TI - Transcriptional regulation of the differentiation-linked human K4 promoter is dependent upon esophageal-specific nuclear factors. AB - The stratified squamous epithelium comprises actively proliferating basal cells that undergo a program of differentiation accompanied by morphological, biochemical, and genetic changes. The transcriptional regulatory signals and the genes that orchestrate this switch from proliferation to differentiation can be studied through the keratin gene family. Given the localization of keratin 4 (K4) to the early differentiated suprabasal compartment and having previously demonstrated that targeted disruption of this gene in murine embryonic stem cells results in impairment of the normal differentiation program in esophageal and corneal epithelial cells, we studied the transcriptional regulation of the human K4 promoter. A panel of K4 promoter deletions were found in transient transfection assays to be predominantly active in esophageal and corneal cell lines. A critical cis-regulatory element resides between -163 and -140 bp and contains an inverted CACACCT motif. A site-directed mutated version of this motif within the K4 promoter renders it inactive, whereas the wild-type version is active in a heterologous promoter system. It specifically binds esophageal specific zinc-dependent transcriptional factors. Our studies demonstrate that regulation of the human K4 promoter is in part mediated through tissue-specific transcriptional factors. PMID- 9727006 TI - Purification and characterization of wild-type and mutant "classical" nitroreductases of Salmonella typhimurium. L33R mutation greatly diminishes binding of FMN to the nitroreductase of S. typhimurium. AB - "Classical" nitroreductase of Salmonella typhimurium is a flavoprotein that catalyzes the reduction of nitroaromatics to metabolites that are toxic, mutagenic, or carcinogenic. This enzyme represents a new class of flavin dependent enzymes, which includes nitroreductases of Enterobacter cloacae and Escherichia coli, flavin oxidoreductase of Vibrio fischeri, and NADH oxidase of Thermus thermophilus. To investigate the structure-function relation of this class of enzymes, the gene encoding a mutant nitroreductase was cloned from S. typhimurium strain TA1538NR, and the enzymatic properties were compared with those of the wild-type. DNA sequence analysis revealed a T to G mutation in the mutant nitroreductase gene, predicting a replacement of leucine 33 with arginine. In contrast to the wild-type enzyme, the purified protein with a mutation of leucine 33 to arginine has no detectable nitroreductase activities in the standard assay conditions and easily lost FMN by dialysis or ultrafiltration. In the presence of an excess amount of FMN, however, the mutant protein exhibited a weak but measurable enzyme activity, and the substrate specificity was similar to that of the wild-type enzyme. Possible mechanisms by which the mutation greatly diminishes binding of FMN to the nitroreductase are discussed. PMID- 9727007 TI - Proliferating and migrating mesangial cells responding to injury express a novel receptor protein-tyrosine phosphatase in experimental mesangial proliferative glomerulonephritis. AB - The mesangial cell provides structural support to the kidney glomerulus. A polymerase chain reaction-based cDNA display approach identified a novel protein tyrosine phosphatase, rPTP-GMC1, whose transcript expression is transiently and dramatically up-regulated during the period of mesangial cell migration and proliferation that follows mesangial cell injury in the anti-Thy 1 model of mesangial proliferative glomerulonephritis in the rat. In situ hybridization analysis confirmed that rPTP-GMC1 mRNA is up-regulated specifically by mesangial cells responding to the injury and is not detectable in other cells in the kidney or in many normal tissues. In cell culture, rPTP-GMC1 is expressed by mesangial cells but not by glomerular endothelial or epithelial cells (podocytes). The longest transcript (7.5 kilobases) encodes a receptor-like protein-tyrosine phosphatase consisting of a single catalytic domain, a transmembrane segment, and 18 fibronectin type III-like repeats in the extracellular segment. A splice variant predicts a truncated molecule missing the catalytic domain. rPTP-GMC1 maps to human chromosome 12q15 and to the distal end of mouse chromosome 10. The predicted structure of rPTP-GMC1 and its pattern of expression in vivo and in culture suggest that it plays a role in regulating the adhesion and migration of mesangial cells in response to injury. PMID- 9727008 TI - The Pad1+ gene encodes a subunit of the 26 S proteasome in fission yeast. AB - We have isolated a fission yeast mutant, mts5-1, in a screen for mutations that confer both methyl 2-benzimidazolecarbamate resistance (MBCR) and temperature sensitivity (ts) on Schizosaccharomyces pombe. This screen has previously isolated mutations in the 26 S proteasome subunits Mts2, Mts3, and Mts4. We show that the mutation in the mts5-1 strain occurs in the pad1(+) gene. pad1(+) was originally isolated on a multicopy plasmid that was capable of conferring staurosporine resistance on a wild type strain. mts5-1/pad1-1 has a similar phenotype to 26 S proteasome mutants previously isolated in the same screen and we show that Pad1 interacts genetically with two of these subunits, Mts3 and Mts4. In this study we describe the identification of Pad1 as a subunit of the 26 S proteasome in fission yeast. PMID- 9727009 TI - Bcl-2 activates the transcription factor NFkappaB through the degradation of the cytoplasmic inhibitor IkappaBalpha. AB - Nuclear factor kappaB (NFkappaB) is a ubiquitously expressed transcription factor that is regulated by the cytoplasmic inhibitor protein IkappaBalpha. Biological agents such as tumor necrosis factor alpha (TNFalpha), which activate NFkappaB, result in the rapid degradation of IkappaBalpha. Adenoviral-mediated gene transfer of Bcl-2 prevents apoptosis of neonatal ventricular myocytes induced by TNFalpha. In view of the growing evidence that NFkappaB may play an important role in regulating apoptosis, we determined whether TNFalpha and Bcl-2 could modulate the activity of NFkappaB in ventricular myocytes. Stimulation of myocytes with TNFalpha resulted in a 2.1-fold increase (p < 0.001) in NFkappaB dependent gene transcription and nuclear DNA binding. Similarly, a 1.9-fold increase (p < 0.0002) in NFkappaB-dependent gene transcription was observed in myocytes expressing Bcl-2. Nuclear DNA binding activity of NFkappaB was significantly increased in myocytes expressing Bcl-2, with a concomitant reduction in IkappaBalpha protein level. The Bcl-2-mediated loss of IkappaBalpha could be prevented by the proteasome inhibitor lactacystin, consistent with the notion that the targeted degradation of IkappaBalpha consequent to overexpression of Bcl-2 utilizes the ubiquitin-proteasome pathway. This was further tested in human 293 cells in which the N-terminal region of IkappaBalpha was identified to be an important regulatory site for Bcl-2. Deletion of this region or a serine to alanine substitution mutant at amino acids 32 and 36, which are defective for both phosphorylation and degradation, were more resistant than wild type IkappaBalpha to the inhibitory effects of Bcl-2. To our knowledge, this provides the first evidence for the regulation of IkappaBalpha by Bcl-2 and suggests a link between Bcl-2 and the NFkappaB signaling pathway in the suppression of apoptosis. PMID- 9727011 TI - Collagenase 2 (MMP-8) expression in murine tissue-remodeling processes. Analysis of its potential role in postpartum involution of the uterus. AB - Neutrophil collagenase or collagenase 2 (MMP-8) is unique among the family of matrix metalloproteinases (MMPs) because of its exclusive pattern of expression in inflammatory conditions. At present, no evidence of the occurrence of this enzyme in tissues other than human has been reported. In this work, we have cloned the murine homologue of human collagenase 2. The isolated cDNA contains an open reading frame coding for a polypeptide of 465 amino acids, which is 74% identical to its human counterpart. The mouse collagenase 2 exhibits the domain structure characteristic of several MMPs, including a signal sequence, a prodomain with the cysteine residue essential for enzyme latency, an activation locus with the Zinc-binding site, and a COOH-terminal fragment with sequence similarity to hemopexin. It also contains the three conserved residues (Tyr-209, Asp-230, and Gly-232) located around the Zinc-binding site and are distinctive of the collagenase subfamily. Northern blot analysis of RNAs isolated from a variety of mouse tissues revealed that collagenase 2 is expressed at late stages during mouse embryogenesis, coinciding with the appearance of hematopoietic cells. In addition, collagenase 2 was highly expressed in the postpartum uterus starting at 1 day postpartum and extending up to 5 days. Enzymatic analysis revealed that matrilysin, another MMP overexpressed in uterine tissue, is able to activate murine procollagenase 2. These data suggest that both enzymes could form an activation cascade resulting in the generation of the collagenolytic activity required during the process of massive connective tissue resumption occurring in the involuting uterus. PMID- 9727010 TI - Identification of a novel ankyrin isoform (AnkG190) in kidney and lung that associates with the plasma membrane and binds alpha-Na, K-ATPase. AB - Ankyrins are a family of adapter molecules that mediate linkages between integral membrane and cytoskeletal proteins. Such interactions are crucial to the polarized distribution of membrane proteins in transporting epithelia. We have cloned and characterized a novel 190-kDa member of this family from a rat kidney cDNA library, which we term AnkG190 based on the predicted size and homology with the larger neuronal AnkG isoform. AnkG190 displays a unique 31-residue amino terminus, a repeats domain consisting of 24 repetitive 33-residue motifs, a spectrin binding domain, and a truncated regulatory domain. Probes derived from the unique amino terminus hybridize to an 8-kilobase message exclusively in kidney and lung and specifically to the kidney outer medullary collecting ducts by in situ hybridization. Transfections of Madin-Darby canine kidney and COS-7 epithelial cell lines with a full-length AnkG190 construct result in (a) expression at the lateral plasma membrane, (b) functional assembly with the cytoskeleton, and (c) interaction with at least one membrane protein, the Na,K ATPase. Two independent Na,K-ATPase binding domains on AnkG190 are demonstrated as follows: one within the distal 12 ankyrin repeats, and a second site within the spectrin binding domain. Thus, ankyrins may interact with integral membrane proteins in a pleiotropic manner that may involve complex tertiary structural determinants. PMID- 9727012 TI - Novel members of the Vesl/Homer family of PDZ proteins that bind metabotropic glutamate receptors. AB - Vesl-1S (186 amino acids, also called Homer) is a protein containing EVH1- and PDZ-like domains whose expression in the hippocampus is regulated during long term potentiation (LTP), one form of synaptic plasticity thought to underlie memory formation (Kato, A., Ozawa, F., Saitoh, Y., Hirai, K., and Inokuchi, K. (1997) FEBS Lett. 412, 183-189; Brakeman, P. R., Lanahan, A. A., O'Brien, R., Roche, K., Barnes, C. A., Huganir, R. L., and Worley, P. F. (1997) Nature 386, 284-288). Here we report additional members of the Vesl/Homer family of proteins, Vesl-1L and Vesl-2. Vesl-1L (366 amino acids), a splicing variant of Vesl-1S, shares N-terminal 175 amino acids with Vesl-1S and contains additional amino acids at the C terminus. Vesl-2 (354 amino acids) was highly related to Vesl-1L in that both contain EVH1- and PDZ-like domains at the N terminus (86% conservation) and an MCC (mutated in colorectal cancer)-like domain and a leucine zipper at the C terminus. In contrast to vesl-1S, we observed no changes in the levels of vesl-1L and vesl-2 mRNAs during dentate gyrus LTP. All these proteins interacted with metabotropic glutamate receptors (mGluR1 and mGluR5) as well as several hippocampal proteins in vitro. Vesl-1L and Vesl-2, but not Vesl-1S, interacted with each other through the C-terminal portion that was absent in Vesl 1S. Vesl-1L and Vesl-2 may mediate clustering of mGluRs at synaptic junctions. We propose that Vesl-1S may be involved in the structural changes that occur at metabotropic glutamatergic synapses during the maintenance phase of LTP by modulating the redistribution of synaptic components. PMID- 9727013 TI - A novel mutation in the switch 3 region of Gsalpha in a patient with Albright hereditary osteodystrophy impairs GDP binding and receptor activation. AB - Albright hereditary osteodystrophy (AHO), a disorder characterized by skeletal abnormalities and obesity, is associated with heterozygous inactivating mutations in the gene for Gsalpha. A novel Gsalpha mutation encoding the substitution of tryptophan for a nonconserved arginine within the switch 3 region (Gsalpha R258W) was identified in an AHO patient. Although reverse transcription-polymerase chain reaction studies demonstrated that mRNA expression from wild type and mutant alleles was similar, Gsalpha expression in erythrocyte membranes from the affected patient was reduced by 50%. A Gsalpha R258W cDNA, as well as one with arginine replaced by alanine (Gsalpha R258A), was generated, and the biochemical properties of in vitro transcription/translation products were examined. When reconstituted with cyc- membranes, both mutant proteins were able to stimulate adenylyl cyclase normally in the presence of guanosine- 5'-O-(3-thiotriphosphate) (GTPgammaS) but had decreased ability in the presence of isoproterenol or AlF4- (a mixture of 10 microM AlCl3 and 10 mM NaF). The ability of each mutant to bind and be activated by GTPgammaS or AlF4- was assessed by trypsin protection assays. Both mutants were protected normally by GTPgammaS but showed reduced protection in the presence of AlF4-. The addition of excess GDP (2 mM) was able to rescue the ability of AlF4- to protect the mutants, suggesting that they might have reduced affinity for GDP. A Gsalpha R258A mutant purified from Escherichia coli had decreased affinity for GDP and an apparent rate of GDP release that was 10 fold greater than that of wild type Gsalpha. Sucrose density gradient analysis demonstrated that both Gsalpha R258W and Gsalpha R258A were thermolabile at higher temperatures and that denaturation of both mutants was prevented by the presence of 0.1 mM GTPgammaS or 2 mM GDP. The crystal structure of Gsalpha demonstrates that Arg258 interacts with a conserved residue in the helical domain (Gln170). Arg258 substitutions would be predicted to open the cleft between the GTPase and helical domains, allowing for increased GDP release in the inactive state, resulting in enhanced thermolability and reduced AlF4--induced adenylyl cyclase stimulation and trypsin protection, since activation by AlF4- requires bound GDP. PMID- 9727014 TI - Characterization of recombinant adrenodoxin reductase homologue (Arh1p) from yeast. Implication in in vitro cytochrome p45011beta monooxygenase system. AB - The mammalian electron transfer chain of mitochondrial cytochrome P450 forms involved in steroidogenesis includes very specific proteins, namely adrenodoxin reductase and adrenodoxin. Adrenodoxin reductase transfers electrons from NADPH to adrenodoxin, which subsequently donates them to the cytochrome P450 forms. The Saccharomyces cerevisiae ARH1 gene product (Arh1p) presents homology to mammalian adrenodoxin reductase. We demonstrate the capacity of recombinant Arh1p, made in Escherichia coli, to substitute for its mammalian homologue in ferricyanide, cytochrome c reduction, and, more importantly, in vitro 11beta-hydroxylase assays. Electrons could be transferred from NADPH and NADH as measured in the cytochrome c reduction assay. Apparent Km values were determined to be 0.5, 0.6, and 0.1 microM for NADPH, NADH, and bovine adrenodoxin, respectively. These values differ slightly from those of mammalian adrenodoxin reductase, except for NADH, which is a very poor electron donor to the mammalian protein. Subcellular fractionation studies have localized Arh1p to the inner membrane of yeast mitochondria. The biological function of Arh1p remains unknown, and to date, no mitochondrial cytochrome P450 has been identified. ARH1 is, however, essential for yeast viability because an ARH1 gene disruption is lethal not only in aerobic growth conditions but also, surprisingly enough, during fermentation. PMID- 9727015 TI - Flagellar motor-switch binding face of CheY and the biochemical basis of suppression by CheY mutants that compensate for motor-switch defects in Escherichia coli. AB - CheY is a response regulator protein of Escherichia coli that interacts with the flagellar motor-switch complex to modulate flagellar rotation during chemotaxis. The switch complex is composed of three proteins, FliG, FliM, and FliN. Recent biochemical data suggest a direct interaction of CheY with FliM. In order to determine the FliM binding face of CheY, we isolated dominant suppressors of fliM mutations in cheY with limited allele specificity. The protein products of suppressor cheY alleles were purified and assayed for FliM binding. Six out of nine CheY mutants were defective in FliM binding. Suppressor amino acid substitutions were mapped on the crystal structure of CheY showing clustering of reduced binding mutations on a solvent-accessible face of CheY, thus revealing a FliM binding face of CheY. To examine the basis of genetic suppression, we cloned, purified, and tested FliM mutants for CheY binding. Like the wild-type FliM, the mutants were also defective in binding to various CheY suppressor mutants. This was not expected if CheY suppressors were compensatory conformational suppressors. Furthermore, a comparison of flagellar rotation patterns indicated that the cheY suppressors had readjusted the clockwise bias of the fliM strains. However, a chemotaxis assay revealed that the readjustment of the clockwise bias was not sufficient to make cells chemotactic. Although the suppressors did not restore chemotaxis, they did increase swarming on motility plates by a process called "pseudotaxis." Therefore, our genetic selection scheme generated suppressors of pseudotaxis or switch bias adjustment. The binding results suggest that the mechanism for this adjustment is the reduction in binding affinity of activated CheY. Therefore, these suppressors identified the switch-binding surface of CheY by loss-of-function defects rather than gain-of function compensatory conformational changes. PMID- 9727016 TI - Real time visualization of agonist-mediated redistribution and internalization of a green fluorescent protein-tagged form of the thyrotropin-releasing hormone receptor. AB - The long isoform of the rat thyrotropin-releasing hormone receptor (TRHR) was modified by the addition of a vesicular stomatitis virus (VSV) epitope tag and green fluorescent protein (GFP). VSV-TRHR-GFP bound TRH with affinity similar to that of the unmodified receptor and stimulated [3H]inositol phosphate production. A clone stably expressing VSV-TRHR-GFP at some 120,000 copies/cell was selected to visualize this receptor during cellular exposure to TRH. Internalization was detected within 3-5 min after treatment with 1 x 10(-7) M TRH, with dramatic reductions in plasma membrane localization achieved within 10-15 min. The TRHR antagonist/inverse agonist chlordiazepoxide competitively inhibited internalization. Hyperosmotic sucrose inhibited internalization of VSV-TRHR-GFP, measured both by intact cell [3H]TRH binding studies and by confocal microscopy. Now TRH caused a redistribution of VSV-TRHR-GFP to highly punctate but plasma membrane-delineated foci. Pretreatment with the microtubule-disrupting agent nocodazole allowed internalization of the VSV-TRHR-GFP construct but only into vesicles that remained in close apposition to the plasma membrane. Covisualization of VSV-TRHR-GFP and Texas Red transferrin initially indicated entirely separate localizations. After exposure to TRH substantial amounts of VSV TRHR-GFP were present in vesicles overlapping those containing Texas Red transferrin. Such results demonstrate the G protein-coupling capacity and provide real time visualization of the processes of internalization of a TRH-receptor-GFP construct in response to agonist. PMID- 9727017 TI - Apoptosis induced by Drosophila reaper and grim in a human system. Attenuation by inhibitor of apoptosis proteins (cIAPs). AB - Previous genetic studies have established Reaper and Grim as central regulators of apoptosis in Drosophila melanogaster. Reaper and Grim induce extensive apoptosis in Drosophila, yet share no homology to known vertebrate proteins. In this study, we show for the first time that ectopic expression of Reaper or Grim induced substantial apoptosis in mammalian cells. Reaper- or Grim-induced apoptosis was inhibited by a broad range of caspase inhibitors and by human inhibitor of apoptosis proteins cIAP1 and cIAP2. Additionally, in vivo binding studies demonstrated that both Reaper and Grim physically interacted with human IAPs through a homologous 15-amino acid N-terminal segment. Deletion of this segment from either Reaper or Grim abolished binding to cIAPs. In vitro binding experiments indicated that Reaper and Grim bound specifically to the BIR domain containing region of cIAPs as deletion of this region resulted in loss of binding. The physical interaction was further confirmed by immunolocalization. When co-expressed, Reaper or Grim co-localized with cIAP1. However, deletion of the N-terminal 15 amino acids of Reaper or Grim abolished co-localization with cIAP1, suggesting that this homologous region can serve as a protein-protein interacting domain in regulating cell death. Moreover, by virtue of this interaction, we demonstrate that cIAPs can regulate Reaper and Grim by abrogating their ability to activate caspases and thereby inhibit apoptosis. This is the first function attributed to this 15-amino acid N-terminal domain that is the only region having significant homology between these Drosophila death inducers. PMID- 9727018 TI - Molecular analysis of human interleukin-9 receptor transcripts in peripheral blood mononuclear cells. Identification of a splice variant encoding for a nonfunctional cell surface receptor. AB - Genetic studies on mouse models of asthma have identified interleukin-9 (IL9) as a determining factor in controlling bronchial hyperresponsiveness, a hallmark of the disease. Recently, the human IL9 receptor (hIL9R) gene locus has also been implicated in determining susceptibility to bronchial hyperresponsiveness and asthma. In order to evaluate the structure and function of the encoded product, we analyzed receptor transcripts derived from peripheral blood mononuclear cells of 50 unrelated donors. Sequence analysis of the entire coding region identified a splice variant that contains an in frame deletion of a single residue at codon 173 (DeltaQ). This variant could be permanently expressed in a cytokine-dependent murine T-cell line but lacked the ability to induce proliferation in response to human IL9. In situ analyses of cells expressing the wild-type and DeltaQ receptors found both forms to be expressed at the cell surface, but the DeltaQ receptor was unable to bind hIL9 and could not be recognized by N-terminal specific antibodies. These findings demonstrate that hIL9RDeltaQ presents an altered structure and function and suggests its potential role in down-regulating IL9 signaling in effector cells and associated biological processes. PMID- 9727019 TI - Inhibitory interactions of the bradykinin B2 receptor with endothelial nitric oxide synthase. AB - It has been shown previously that the endothelial nitric-oxide synthase (eNOS) interacts reversibly with the plasmalemmal caveolae structural protein, caveolin 1. The eNOS-caveolin-1 interaction inhibits eNOS catalytic activity. In the present study, we show that eNOS also participates in reversible inhibitory interactions with the G protein-coupled bradykinin B2 receptor. eNOS and the B2 receptor are coimmunoprecipitated from endothelial cell lysates by antibodies directed against either of the two proteins. A glutathione S-transferase fusion protein containing intracellular domain 4 of the receptor is bound by purified recombinant eNOS in in vitro binding assays. The fusion protein selectively inhibits the activity of purified eNOS. A synthetic peptide corresponding to membrane-proximal residues 310-334 in intracellular domain 4 also potently inhibits eNOS activity (IC50 < 1 microM). Treatment of cultured endothelial cells with bradykinin or Ca2+ ionophore promotes a rapid dissociation of the eNOS.B2 receptor complex. These data demonstrate that the bradykinin B2 receptor physically associates with eNOS in a ligand- and Ca2+-dependent manner. Reversible and inhibitory membrane-docking interactions of eNOS, therefore, are not restricted to those with caveolin-1 but also occur with the bradykinin B2 receptor. PMID- 9727020 TI - Enhanced binding of altered H-NS protein to flagellar rotor protein FliG causes increased flagellar rotational speed and hypermotility in Escherichia coli. AB - H-NS is an Escherichia coli nucleoid protein known only to function as a modulator of gene expression. In this study, we found that specific single amino acid substitutions in H-NS caused an approximately 50% increase in flagellum rotational speed. In fluorescence anisotropy and chemical cross-linking assays, H NS interacted with the flagellar torque-generating rotor protein FliG to form a complex with a Kd of 2.15 microM. Furthermore, one of the altered H-NS proteins that exhibited high speed flagellum rotation bound FliG 50% tighter than wild type H-NS. These results demonstrate the first non-regulatory role for H-NS and provide a direct correlation between H-NS-FliG binding affinities, flagellar rotation, and motor torque generation. PMID- 9727021 TI - Mechanical stretch induces hypertrophic responses in cardiac myocytes of angiotensin II type 1a receptor knockout mice. AB - Many lines of evidence have suggested that angiotensin II (AngII) plays an important role in the development of cardiac hypertrophy through AngII type 1 receptor (AT1). To determine whether AngII is indispensable for the development of mechanical stress-induced cardiac hypertrophy, we examined the activity of mitogen-activated protein kinase (MAPK) family and the expression of the c-fos gene as hypertrophic responses after stretching cultured cardiac myocytes of AT1a knockout (KO) mice. When cardiac myocytes were stretched by 20% for 10 min, extracellular signal-regulated protein kinases (ERKs) were strongly activated in KO cardiomyocytes as well as wild type (WT) myocytes. Both basal and stimulated levels of ERKs were higher in cardiomyocytes of KO mice than in those of WT mice. Activation of another member of the MAPK family, p38(MAPK), and expression of the c-fos gene were also induced by stretching cardiac myocytes of both types of mice. An AT1 antagonist attenuated stretch-induced activation of ERKs in WT cardiomyocytes but not in KO cardiomyocytes. Down-regulation of protein kinase C inhibited stretch-induced ERK activation in WT cardiomyocytes, whereas a broad spectrum tyrosine kinase inhibitor (genistein) and selective inhibitors of epidermal growth factor receptor (tyrphostin, AG1478, and B42) suppressed stretch induced activation of ERKs in KO cardiac myocytes. Epidermal growth factor receptor was phosphorylated at tyrosine residues by stretching cardiac myocytes of KO mice. These results suggest that mechanical stretch could evoke hypertrophic responses in cardiac myocytes that lack the AT1 signaling pathway possibly through tyrosine kinase activation. PMID- 9727022 TI - A new metabolic link between fatty acid de novo synthesis and polyhydroxyalkanoic acid synthesis. The PHAG gene from Pseudomonas putida KT2440 encodes a 3 hydroxyacyl-acyl carrier protein-coenzyme a transferase. AB - To investigate the metabolic link between fatty acid de novo synthesis and polyhydroxyalkanoic acid (PHA) synthesis, we isolated mutants of Pseudomonas putida KT2440 deficient in this metabolic route. The gene phaG was cloned by phenotypic complementation of these mutants; it encoded a protein of 295 amino acids with a molecular mass of 33,876 Da, and the amino acid sequence exhibited 44% amino acid identity to the primary structure of the rhlA gene product, which is involved in the rhamnolipid biosynthesis in Pseudomonas aeruginosa PG201. S1 nuclease protection assay identified the transcriptional start site 239 base pairs upstream of the putative translational start codon. Transcriptional induction of phaG was observed when gluconate was provided, and PHA synthesis occurred from this carbon source. No complementation of the rhlA mutant P. aeruginosa UO299-harboring plasmid pBHR81, expressing phaG gene under lac promoter control, was obtained. Heterologous expression of phaG in Pseudomonas oleovorans, which is not capable of PHA synthesis from gluconate, enabled PHA synthesis on gluconate as the carbon source. Native recombinant PhaG was purified by native polyacrylamide gel electrophoresis from P. oleovorans-harboring plasmid pBHR81. It catalyzes the transfer of the acyl moiety from in vitro synthesized 3 hydroxydecanoyl-CoA to acyl carrier protein, indicating that PhaG exhibits a 3 hydroxyacyl-CoA-acyl carrier protein transferase activity. PMID- 9727023 TI - Ras isoforms vary in their ability to activate Raf-1 and phosphoinositide 3 kinase. AB - Ha-, N-, and Ki-Ras are ubiquitously expressed in mammalian cells and can all interact with the same set of effector proteins. We show here, however, that in vivo there are marked quantitative differences in the ability of Ki- and Ha-Ras to activate Raf-1 and phosphoinositide 3-kinase. Thus, Ki-Ras both recruits Raf-1 to the plasma membrane more efficiently than Ha-Ras and is a more potent activator of membrane-recruited Raf-1 than Ha-Ras. In contrast, Ha-Ras is a more potent activator of phosphoinositide 3-kinase than Ki-Ras. Interestingly, the ability of Ha-Ras to recruit Raf-1 to the plasma membrane is significantly increased when the Ha-Ras hypervariable region is shortened so that the spacing of the Ha-Ras GTPase domains from the inner surface of the plasma membrane mimicks that of Ki-Ras. Importantly, these data show for the first time that the activation of different Ras isoforms can have distinct biochemical consequences for the cell. The mutation of specific Ras isoforms in different human tumors can, therefore, also be rationalized. PMID- 9727024 TI - p35, the neuronal-specific activator of cyclin-dependent kinase 5 (Cdk5) is degraded by the ubiquitin-proteasome pathway. AB - Cyclin-dependent kinase 5 (Cdk5) was originally isolated by its close homology to the human CDC2 gene, which is a key regulator of cell cycle progression. However, unlike other Cdks, the activity of Cdk5 is required in post-mitotic neurons. The neuronal-specific p35 protein, which shares no homology to cyclins, was identified by virtue of its association and activation of Cdk5. Gene targeting studies in mice have shown that the p35/Cdk5 kinase is required for the proper neuronal migration and development of the mammalian cortex. We have investigated the regulation of the p35/Cdk5 kinase. Here we show that p35, the activator of Cdk5, is a short-lived protein with a half-life (t1/2) of 20 to 30 min. Specific proteasome inhibitors such as lactacystin greatly stabilize p35 in vivo. Ubiquitination of p35 can be readily demonstrated in vitro and in vivo. Inhibition of Cdk5 activity by a specific Cdk inhibitor, roscovitine, or by overexpression of a dominant negative mutant of Cdk5 increases the stability of p35 by 2- to 3-fold. Furthermore, phosphorylation mutants of p35 also stabilize p35 2- to 3-fold. Together, these observations demonstrate that the p35/Cdk5 kinase can be subject to rapid turnover in vivo and suggest that phosphorylation of p35 upon Cdk5 kinase activation plays a autoregulatory role in p35 degradation mediated by ubiquitin-mediated proteolysis. PMID- 9727025 TI - AP1 proteins mediate the cAMP response of the dopamine beta-hydroxylase gene. AB - Neurotransmitter biosynthesis is regulated by environmental stimuli, which transmit intracellular signals via second messengers and protein kinase pathways. For the catecholamine biosynthetic enzymes, dopamine beta-hydroxylase and tyrosine hydroxylase, regulation of gene expression by cyclic AMP, diacyl glycerol, and Ca2+ leads to increased neurotransmitter biosynthesis. In this report, we demonstrate that the cAMP-mediated regulation of transcription from the dopamine beta-hydroxylase promoter is mediated by the AP1 proteins c-Fos, c Jun, and JunD. Following treatment of cultured cells with cAMP, protein complexes bound to the dopamine beta-hydroxylase AP1/cAMP response element element change from consisting of c-Jun and JunD to include c-Fos, c-Jun, and JunD. The homeodomain protein Arix is also a component of this DNA-protein complex, binding to the adjacent homeodomain recognition sites. Transfection of a dominant negative JunD expression plasmid inhibits cAMP-mediated expression of the dopamine beta-hydroxylase promoter construct in PC12 and CATH.a cells. In addition to the role of c-Fos in regulating dopamine beta-hydroxylase gene expression in response to cAMP, a second pathway, involving Rap1/B-Raf is involved. These experiments illustrate an unusual divergence of cAMP-dependent protein kinase signaling through multiple pathways that then reconverge on a single element in the dopamine beta-hydroxylase promoter to elicit activation of gene expression. PMID- 9727026 TI - Impaired mismatch extension by a herpes simplex DNA polymerase mutant with an editing nuclease defect. AB - The D368A mutation within the 3'-5'-exonuclease domain of the herpes simplex type 1 DNA polymerase inactivates this nuclease and severely interferes with virus viability. Compared with the wild type enzyme, the D368A mutant exhibits substantially elevated rates of incorrect nucleotide incorporation, as measured in a LacZ reversion assay. This high rate occurs in the presence of high levels of dNTPs, a condition that forces the enzyme to extend mismatched primers. Hence, the mutant fails to correct many misincorporations that are removed in the wild type. In addition, the mutant shows a much reduced ability to replicate DNA templates primed with a 3'-mismatch as compared with wild type. This extension defect also appears more severe than observed for replicases which naturally lack editing nucleases. Based on these findings, we suggest that the inability of the D368A herpes simplex mutant polymerase to replicate beyond a mismatched base pair severely inhibits viral replication. PMID- 9727027 TI - Functional characterization of a plant importin alpha homologue. Nuclear localization signal (NLS)-selective binding and mediation of nuclear import of nls proteins in vitro. AB - Nuclear import of most nuclear proteins is initiated by recognition of the nuclear localization signal (NLS) by importin alpha. We recently isolated an importin alpha homologue from rice (rice importin alpha1) and demonstrated that transcription of the gene is down-regulated by light in rice leaves. To address the function of rice importin alpha1 in the process of nuclear import of proteins, we performed in vitro binding and nuclear import assays. The rice importin alpha1 showed specific binding to fusion proteins containing either monopartite or bipartite NLSs, but not to a fusion protein containing a Matalpha 2-type NLS, suggesting that there exists selective binding of rice importin alpha1 to different plant NLSs. The rice importin alpha1 is also capable of forming a complex with mouse importin beta and NLS protein in vitro. An in vitro nuclear import assay using permeabilized HeLa cells revealed that rice importin alpha1, in conjunction with other vertebrate transport factors, mediates the nuclear envelope docking of NLS proteins and their subsequent translocation into the nucleus. These data provide strong, direct evidence suggesting that rice importin alpha1 functions as a component of the NLS receptor in plant cells. PMID- 9727028 TI - Structural maintenance of chromosomes protein C-terminal domains bind preferentially to DNA with secondary structure. AB - Structural maintenance of chromosomes (SMC) proteins interact with DNA in chromosome condensation, sister chromatid cohesion, DNA recombination, and gene dosage compensation. How individual SMC proteins and their functional domains bind DNA has not been described. We demonstrate the ability of the C-terminal domains of Saccharomyces cerevisiae SMC1 and SMC2 proteins, representing two major subfamilies with different functions, to bind DNA in an ATP-independent manner. Three levels of DNA binding specificity were observed: 1) a >100-fold preference for double-stranded versus single-stranded DNA; 2) a high affinity for DNA fragments able to form secondary structures and for synthetic cruciform DNA molecules; and 3) a strong preference for AT-rich DNA fragments of particular types. These include fragments from the scaffold-associated regions, and an alternating poly(dA-dT)-poly(dT-dA) synthetic polymer, as opposed to a variety of other polymers. Reannealing of complementary DNA strands is also promoted primarily by the C-terminal domains. Consistent with their in vitro DNA binding activity, we show that overexpression of the SMC C termini increases plasmid loss without altering viability or cell cycle progression. PMID- 9727030 TI - Glucose repression in Saccharomyces cerevisiae is related to the glucose concentration rather than the glucose flux. AB - Glucose plays an important regulatory role in the yeast Saccharomyces cerevisiae, which is mostly reflected at the transcriptional level by glucose repression. The signal that initiates glucose repression is unknown, but data indicate that it is located at or above the level of glucose 6-phosphate, suggesting the involvement of either the intracellular or extracellular glucose concentration or the glucose flux in triggering glucose repression. We have investigated the role of the glucose flux and the extracellular glucose concentration in glucose repression by growing the cells in continuous culture under nitrogen limitation. By a step-wise increase in the glucose feed concentration, the glucose flux and extracellular glucose concentrations were modulated in an accurate way. Furthermore, the glucose flux and glucose concentrations were modulated independently of each other by increasing the dilution rate or by the use of fructose as a substrate. Using these approaches we demonstrate that glucose repression is related to the extracellular (or intracellular) glucose concentration rather than the glucose flux. At external glucose concentrations lower than 14 mM, glucose repression of SUC2 gene transcription was not triggered, whereas glucose repression of this gene was activated when the glucose concentration exceeded 18 mM. A comparable effect was observed for the glucose-repressible carbon source fructose. PMID- 9727029 TI - Interaction of human suppressor of cytokine signaling (SOCS)-2 with the insulin like growth factor-I receptor. AB - SOCS (suppressor of cytokine signaling) proteins have been shown to be negative regulators of cytokine receptor signaling via the Janus kinase (JAK)/signal transducer and activator of transcription (STAT) pathway. We have cloned a member of this family (hSOCS-2) by utilizing the insulin-like growth factor I receptor (IGF-IR) cytoplasmic domain as bait in a yeast two-hybrid screen of a human fetal brain library. The hSOCS-2 protein interacted strongly with the activated IGF-IR and not with a kinase negative mutant receptor in the two-hybrid assay. Mutation of receptor tyrosines 950, 1250, 1251, and 1316 to phenylalanine or deletion of the COOH-terminal 93 amino acids did not result in decreased interaction of the receptor with hSOCS-2 protein. hSOCS-1 protein also interacted strongly with IGF IR in the two-hybrid assay. Glutathione S-transferase-hSOCS-2 associated with activated IGF-IR in lysates of mouse fibroblasts overexpressing IGF-IR. Human embryonic kidney cells (293) were transiently transfected with vectors containing IGF-IR and FLAG epitope-tagged hSOCS-2. After IGF-I stimulation, activated IGF-IR was found in anti-FLAG immunoprecipitates and, conversely, FLAG-hSOCS-2 was found in anti IGF-IR immunoprecipitates. Thus, hSOCS-2 interacted with IGF-IR both in vitro and in vivo. HSOCS-2 mRNA was expressed in many human fetal and adult tissues with particularly high abundance in fetal kidney and adult heart, skeletal muscle, pancreas, and liver. These results raise the possibility that SOCS proteins may also play a regulatory role in IGF-I receptor signaling. PMID- 9727031 TI - Mitotic Raf-1 is stimulated independently of Ras and is active in the cytoplasm. AB - Raf-1 is a Ser/Thr protein kinase that is involved in regulation of proliferation, differentiation, and apoptosis. Recently, we and others showed that Raf-1 is not only activated in mitogenic pathways leading to cell cycle entry but also during mitosis. Transient expression studies in COS cells now demonstrate that, in contrast to growth factor-dependent activation of Raf-1, mitotic activation of Raf-1 is Ras-independent. Dominant negative RasS17N does not interfere with mitotic activation of Raf-1, whereas epidermal growth factor dependent stimulation of Raf-1 is inhibited. In addition, the Raf-1 mutant RafR89L, which cannot bind to activated Ras, is still stimulated in mitotic cells. Mitotic activation of Raf-1 seems to be partially dependent on tyrosine phosphorylation since the kinase activity of the Raf mutant RafYY340/341FF, which can no longer be activated by Src, is reduced in mitotic cells. Surprisingly, cell fractionation experiments showed that mitotic-activated Raf-1 is predominantly located in the cytoplasm in contrast to the mitogen-activated Raf-1 that is bound to the plasma membrane. In addition, mitotic activation of Raf-1 does not lead to stimulation of the mitogen-activated protein kinase kinase (MAPKK or MEK) and the extracellular signal-regulated protein kinase (ERK). These data demonstrate that in mitotic cells a Ras-independent mechanism results in a cytoplasmic active Raf-1 kinase which does not signal via the MEK/ERK pathway. These data demonstrate that in mitotic cells a Ras-independent mechanism results in a cytoplasmic active Raf-1 kinase which does not signal via the MEK/ERK pathway. PMID- 9727032 TI - Evidence that protein kinase Cepsilon mediates phorbol ester inhibition of calphostin C- and tumor necrosis factor-alpha-induced apoptosis in U937 histiocytic lymphoma cells. AB - Protein kinase C (PKC) activators, such as the tumor-promoting phorbol esters, have been reported to protect several cell lines from apoptosis induced by a variety of agents. Recent evidence suggests that PKCepsilon is involved in protection of cardiac myocytes from hypoxia-induced cell death (Gray, M. O., Karliner, J. S., and Mochly-Rosen, D. (1997) J. Biol. Chem. 272, 30945-30951). We investigated the protective effects of the phorbol ester 12-O tetradecanoylphorbol-13-acetate (TPA) on U937 histiocytic lymphoma cells induced to undergo apoptosis by tumor necrosis factor-alpha (TNF-alpha) or by the specific PKC inhibitor calphostin C. U937 cells were transiently permeabilized with a peptide (epsilonV1-2) derived from the V1 region of PKCepsilon that has been reported to specifically block translocation of PKCepsilon. The epsilonV1-2 peptide blocked the inhibitory effect of TPA on both TNF-alpha- and calphostin C induced apoptosis. A scrambled version of epsilonV1-2 and a peptide reported to inhibit PKCbeta translocation (betaC2-4) had no effect on the ability of TPA to inhibit apoptosis. These results suggest that PKCepsilon is required for the protective effect of TPA in TNF-alpha- and calphostin C-induced apoptosis. Furthermore, calphostin C reduced membrane-associated PKCepsilon activity and immunoreactivity, suggesting that PKCepsilon may play an important role in leukemic cell survival. PMID- 9727033 TI - Newly identified Chinese hamster ovary cell mutants are defective in biogenesis of peroxisomal membrane vesicles (Peroxisomal ghosts), representing a novel complementation group in mammals. AB - We isolated peroxisome biogenesis-defective mutants from Chinese hamster ovary cells by the 9-(1'-pyrene)nonanol/ultraviolet (P9OH/UV) method. Seven cell mutants, ZP116, ZP119, ZP160, ZP161, ZP162, ZP164, and ZP165, of 11 P9OH/UV resistant cell clones showed cytosolic localization of catalase, a peroxisomal matrix enzyme, apparently indicating a defect of peroxisome biogenesis. By transfection of PEX cDNAs and cell fusion analysis, mutants ZP119 and ZP165 were found to belong to a novel complementation group (CG), distinct from earlier mutants. CG analysis by cell fusion with fibroblasts from patients with peroxisome biogenesis disorders such as Zellweger syndrome indicated that ZP119 and ZP165 were in the same CG as the most recently identified human CG-J. The peroxisomal matrix proteins examined, including PTS1 proteins as well as a PTS2 protein, 3-ketoacyl-CoA thiolase, were also found in the cytosol in ZP119 and ZP165. Furthermore, these mutants showed typical peroxisome assembly-defective phenotype such as severe loss of resistance to 12-(1'-pyrene)dodecanoic acid/UV treatment. Most strikingly, peroxisomal reminiscent vesicular structures, so called peroxisomal ghosts noted in all CGs of earlier Chinese hamster ovary cell mutants as well as in eight CGs of patients' fibroblasts, were not discernible in ZP119 and ZP165, despite normal synthesis of peroxisomal membrane proteins. Accordingly, ZP119 and ZP165 are the first cell mutants defective in import of both soluble and membrane proteins, representing the 14th peroxisome-deficient CG in mammals, including humans. PMID- 9727034 TI - A novel component involved in ubiquitination is required for development of Dictyostelium discoideum. AB - A novel component of the ubiquitination system, called NOSA, is essential for cellular differentiation in Dictyostelium discoideum. Disruption of nosA does not affect the growth rate but causes an arrest in development after the cells have aggregated. nosA contains seven exons and codes for a developmentally regulated 3.5-kb mRNA. The 125-kDa NOSA protein is present in the cytosol at constant levels during growth and development. The C-terminal region of NOSA has homology with ubiquitin fusion degradation protein-2 (UFD2) of Saccharomyces cerevisiae and putative homologs in Caenorhabditis elegans and humans. UFD2 is involved in the ubiquitin-mediated degradation of model substrates in which ubiquitin forms part of the translation product, but ufd2 mutants have no detected phenotype. In accord with the homology to UFD2, we found differences in the ubiquitination patterns between nosA mutants and their parental cell line. While general in vivo and in vitro ubiquitination is minimally affected, ubiquitination of individual proteins is altered throughout growth and development in nosA mutants. These findings suggest that events involving ubiquitination are critical for progression through the aggregate stage of the Dictyostelium life cycle. PMID- 9727035 TI - Differential heparin sensitivity of alpha-dystroglycan binding to laminins expressed in normal and dy/dy mouse skeletal muscle. AB - The alpha-dystroglycan binding properties of laminins extracted from fully differentiated skeletal muscle were characterized. We observed that the laminins expressed predominantly in normal adult rat or mouse skeletal muscle bound alpha dystroglycan in a Ca2+-dependent, ionic strength-sensitive, but heparin insensitive manner as we had observed previously with purified placental merosin (Pall, E. A., Bolton, K. M., and Ervasti, J. M. 1996 J. Biol. Chem. 271, 3817 3821). Rat skeletal muscle laminins partially purified by heparin-agarose affinity chromatography also bound alpha-dystroglycan without sensitivity to heparin. We also confirm previous studies of dystrophic dy/dy mouse skeletal muscle showing that the alpha2 chain of merosin is reduced markedly and that the laminin alpha1 chain is not up-regulated detectably. However, we further observed a quantitative decrease in the expression of laminin beta/gamma chain immunoreactivity in alpha2 chain-deficient dy/dy skeletal muscle and reduced alpha-dystroglycan binding activity in laminin extracts from dy/dy muscle. Most interestingly, the alpha-dystroglycan binding activity of residual laminins expressed in merosin-deficient dy/dy skeletal muscle was inhibited dramatically (69 +/- 19%) by heparin. These results identify a potentially important biochemical difference between the laminins expressed in normal and dy/dy skeletal muscle which may provide a molecular basis for the inability of other laminin variants to compensate fully for the deficiency of merosin in some forms of muscular dystrophy. PMID- 9727036 TI - The repression of hormone-activated PEPCK gene expression by glucose is insulin independent but requires glucose metabolism. AB - Phosphoenolpyruvate carboxykinase (PEPCK) is a rate-controlling enzyme in hepatic gluconeogenesis, and it therefore plays a central role in glucose homeostasis. The rate of transcription of the PEPCK gene is increased by glucagon (via cAMP) and glucocorticoids and is inhibited by insulin. Under certain circumstances glucose also decreases PEPCK gene expression, but the mechanism of this effect is poorly understood. The glucose-mediated stimulation of a number of glycolytic and lipogenic genes requires the expression of glucokinase (GK) and increased glucose metabolism. HL1C rat hepatoma cells are a stably transfected line of H4IIE rat hepatoma cells that express a PEPCK promoter-chloramphenicol acetyltransferase fusion gene that is regulated in the same manner as the endogenous PEPCK gene. These cells do not express GK and do not normally exhibit a response of either the endogenous PEPCK gene, or of the trans-gene, to glucose. A recombinant adenovirus that directs the expression of glucokinase (AdCMV-GK) was used to increase glucose metabolism in HL1C cells to test whether increased glucose flux is also required for the repression of PEPCK gene expression. In AdCMV-GK-treated cells glucose strongly inhibits hormone-activated transcription of the endogenous PEPCK gene and of the expressed fusion gene. The glucose effect on PEPCK gene promoter activity is blocked by 5 mM mannoheptulose, a specific inhibitor of GK activity. The glucose analog, 2-deoxyglucose mimics the glucose response, but this effect does not require GK expression. 3-O-methylglucose is ineffective. Glucose exerts its effect on the PEPCK gene within 4 h, at physiologic concentrations, and with an EC50 of 6.5 mM, which approximates the Km of glucokinase. The effects of glucose and insulin on PEPCK gene expression are additive, but only at suboptimal concentrations of both agents. The results of these studies demonstrate that, by inhibiting PEPCK gene transcription, glucose participates in a feedback control loop that governs its production from gluconeogenesis. PMID- 9727037 TI - The domain structure of human receptor-associated protein. Protease sensitivity and guanidine HCl denaturation. AB - The 39-kDa receptor-associated protein (RAP), a specialized chaperone for endocytic receptors of the low density lipoprotein receptor gene family, is a triplicate repeat sequence (residues 1-100, 101-200, and 201-323, respectively), with the three repeats having different functional roles. The goal of the present study was to use a combination of protease sensitivity and guanidine denaturation analyses to investigate whether human RAP correspondingly contained multiple structural domains. Protease sensitivity analysis using six proteolytic enzymes of varying specificity showed that RAP has two protease-resistant regions contained within repeat 1 (residues 15-94) and repeat 3 (residues 223-323). Guanidine denaturation analysis showed that RAP has two phases in its denaturation, an early denaturation transition at 0.6 M guanidine HCl, and a broad second transition between 1.0 and 3.0 M guanidine HCl. Analysis of the denaturation of the individual repeats showed that, despite the similarity in sequence and protease sensitivity between repeats 1 and 3, repeat 1 was a stable structure, with a sharp transition midpoint at 2.4 M guanidine HCl, while repeat 3 was relatively unstable, with a transition midpoint at 0.6 M guanidine HCl. Repeat 2 had a denaturation profile almost identical to that of repeat 3. Denaturation analysis of the contiguous repeats 1 and 2 (residues 1-210) indicated that repeats 1 and 2 probably interact to form one structural domain represented by the broad transition, while repeat 3 constitutes a separate domain represented by the early transition. A two-domain model of RAP three-dimensional structure is proposed that integrates both structural and functional information, in which a helical segment from repeat 2 interacts with the known three-helix bundle of repeat 1 to form a four-helix bundle structural domain, while repeat 3 forms the other structural domain. PMID- 9727038 TI - Subunit interactions in the mammalian alpha-ketoglutarate dehydrogenase complex. Evidence for direct association of the alpha-ketoglutarate dehydrogenase and dihydrolipoamide dehydrogenase components. AB - Selective tryptic proteolysis of the mammalian alpha-ketoglutarate dehydrogenase complex (OGDC) leads to its rapid inactivation as a result of a single cleavage within the N-terminal region of its alpha-ketoglutarate dehydrogenase (E1) component, which promotes the dissociation of the dihydrolipoamide dehydrogenase (E3) enzyme and also a fully active E1' fragment. Similarities between the N terminal region of E1 and the dihydrolipoamide acetyltransferase (E2) and E3 binding components (E3BP) of the pyruvate dehydrogenase complex are highlighted by the specific cross-reactivities of subunit-specific antisera. Analysis of the pattern of release of E1 and E1' polypeptides from the OGDC during tryptic inactivation suggests that both polypeptide chains of individual E1 homodimers must be cleaved to permit the dissociation of the E1 and E3 components. A new protocol has been devised that promotes E1 dissociation from the oligomeric dihydrolipoamide succinyltransferase (E2) core in an active state. Significant levels of overall OGDC reconstitution could also be achieved by re-mixing the constituent enzymes in stoichiometric amounts. Moreover, a high affinity interaction has been demonstrated between the homodimeric E1 and E3 components, which form a stable subcomplex comprising single copies of these two enzymes. PMID- 9727039 TI - DNA-based positive control mutants in the binding site sequence of 434 repressor. AB - As detected by chemical nuclease treatments, the conformation of the 434 repressor-DNA complex depends on the sequence of the bound DNA (Bell, A. C., and Koudelka, G. B. (1993) J. Mol. Biol. 234, 542-553). We show here that these DNA sequence-dependent conformational changes alter the efficiency with which the repressor activates transcription from 434 PRM. Several lines of evidence suggest that binding site sequence affects the repressor's ability to activate transcription by altering the accessibility of the activation surface on the repressor to RNA polymerase. The results presented here show that in addition to affecting transcription by altering the overall binding affinity of protein for DNA, DNA sequence may also modulate the activity of the DNA-bound protein. PMID- 9727040 TI - A role for the p38 mitogen-activated protein kinase/Hsp 27 pathway in cholecystokinin-induced changes in the actin cytoskeleton in rat pancreatic acini. AB - Cholecystokinin (CCK) and other pancreatic secretagogues have recently been shown to activate signaling kinase cascades in pancreatic acinar cells, leading to the activation of extracellular signal-regulated kinases and Jun N-terminal kinases. We now show the presence of a third kinase cascade activating p38 mitogen activated protein (MAP) kinase in isolated rat pancreatic acini. CCK and osmotic stress induced by sorbitol activated p38 MAP kinase within minutes; their effects were dose-dependent, with maximal activation of 2.8- and 4.4-fold, respectively. The effects of carbachol and bombesin on p38 MAP kinase activity were similar to those of CCK, whereas phorbol ester, epidermal growth factor, and vasoactive intestinal polypeptide stimulated p38 MAP kinase by 2-fold or less. Both CCK and sorbitol also increased the tyrosyl phosphorylation of p38 MAP kinase. Using the specific inhibitor of p38 MAP kinase, SB 203580, we found that p38 MAP kinase activity was required for MAP kinase-activated protein kinase-2 activation in pancreatic acini. SB 203580 reduced the level of basal phosphorylation and blocked the increased phosphorylation of Hsp 27 after stimulation with either CCK or sorbitol. CCK treatment induced an initial rapid decrease in total F-actin content of acini, followed by an increase after 40 min. Preincubation with SB 203580 significantly inhibited these changes in F-actin content. Staining of the actin cytoskeleton with rhodamine-conjugated phalloidin and analysis by confocal fluorescence microscopy showed disruption of the actin cytoskeleton after 10 and 40 min of CCK stimulation. Pretreatment with SB 203580 reduced these changes. These findings demonstrate that the activation of p38 MAP kinase is involved not only in response to stress, but also in physiological signaling by gastrointestinal hormones such as CCK, where activation of Gq-coupled receptors stimulates a cascade in which p38 MAP kinase activates MAP kinase-activated protein kinase-2, resulting in Hsp 27 phosphorylation. Activation of p38 MAP kinase, most likely through phosphorylation of Hsp 27, plays a role in the organization of the actin cytoskeleton in pancreatic acini. PMID- 9727041 TI - High constitutive activity of the human formyl peptide receptor. AB - The formyl peptide receptor (FPR) couples to pertussis toxin (PTX)-sensitive Gi proteins to activate chemotaxis and exocytosis in neutrophils. PTX reduces not only formyl peptide-stimulated but also agonist-independent ("basal") Gi-protein activity, suggesting that the FPR is constitutively active. We aimed at identifying an inverse FPR agonist, i.e. a compound that suppresses constitutive FPR activity. In Sf9 insect cell membranes, the G-protein heterotrimer Gialpha2beta1gamma2 reconstituted N-formyl-L-methionyl-L-leucyl-L-phenylalanine (FMLP)-stimulated guanosine 5'-O-(3-thiotriphosphate) (GTPgammaS) binding and GTPgammaS-sensitive high affinity [3H]FMLP binding. The FPR "antagonist" cyclosporin H (CsH) potently and efficiently reduced basal GTPgammaS binding in Sf9 membranes. Another FPR antagonist, N-t-butoxycarbonyl-L-phenylalanyl-L-leucyl L-phenylalanyl-L-leucyl-L- phenylalanine did not inhibit basal GTPgammaS binding but blocked the inhibitory effect of CsH on GTPgammaS binding. Na+ reduced basal GTPgammaS binding and eliminated the inhibitory effect of CsH. Similar effects of FMLP, CsH, and Na+ as in Sf9 membranes were observed with FPR expressed in the mammalian cell line HEK293. Our data show that the human FPR possesses high constitutive activity. CsH is an inverse FPR agonist and stabilizes the FPR in an inactive state. Na+ also stabilizes the FPR in an inactive state and, thereby, diminishes inverse agonist efficacy. PMID- 9727042 TI - A preferred target DNA structure for retroviral integrase in vitro. AB - The retroviral integrase protein catalyzes the insertion of linear viral DNA ends into the host cell DNA. Although integration in vivo is not site-specific, the detection of local and regional preferences within cellular DNA suggests that the integration reaction can be influenced by specific features of host DNA or chromatin. Here we describe highly preferred in vitro integration sites for avian sarcoma virus and human immunodeficiency virus-1 integrases within the stems of plasmid DNA cruciform structures. The preferred sites are adjacent to the loops in the cruciform and are strand-specific. We suggest that the observed preference is due to the end-like character of the stem loop structure that allows DNA unpairing. From these results we propose that such unpairing may enhance both the processing and the joining steps in the integration reaction, and perhaps other cellular recombination reactions as well. PMID- 9727043 TI - Targeting of G protein-coupled receptors to the basolateral surface of polarized renal epithelial cells involves multiple, non-contiguous structural signals. AB - Truncations and chimeras of the alpha2A-adrenergic receptor (alpha2AAR) were evaluated to identify membrane domains responsible for its direct basolateral targeting in Madin-Darby canine kidney cells. An alpha2AAR truncation, encoding transmembrane (TM) regions 1-5, was first delivered basolaterally, but within minutes appeared apically, and at steady-state was primarily lateral in its immunocytochemical localization. A TM 1-5 truncation with the third intracellular loop revealed more intense lateral localization than for the TM 1-5 structure, consistent with the role of the third intracellular loop in alpha2AAR stabilization. Addition of TM 6-7 of A1 adenosine receptor (A1AdoR) to alpha2AARTM1-5 creates a chimera, alpha2AARTM1-5/A1AdoRTM6-7, which was first delivered apically, resulting either from loss of alpha2AAR sorting information in TM 6-7 or acquisition of apical trafficking signals within A1AdoRTM6-7. Evidence that alpha2AARTM6-7 imparts basolateral targeting information is revealed by the significant basolateral localization of the A1AdoRTM1 5/alpha2AARTM6-7 and A1AdoRTM1-5/alpha2AARTM6-7+i3 chimeras, in contrast to the dominant apical localization of A1AdoR. These results reveal that sequences within TM 1-5 and within TM 6-7 of the alpha2AAR confer basolateral targeting, providing the first evidence that alpha2AAR basolateral localization is not conferred by a single region but by non-contiguous membrane-embedded or proximal sequences. PMID- 9727044 TI - Lung endothelial dipeptidyl peptidase IV promotes adhesion and metastasis of rat breast cancer cells via tumor cell surface-associated fibronectin. AB - Endothelial cell adhesion molecules are partly responsible for the distinct organ distribution of cancer metastases. Dipeptidyl peptidase IV (DPP IV) expressed on rat lung capillary endothelia is shown here to be an adhesion receptor for rat breast cancer cells and to mediate lung colonization by these tumor cells. Fibronectin (FN) assembled on breast cancer cell surfaces into multiple, randomly dispersed globules from cellular and plasma FN is identified as the principal ligand for DPP IV. Ligand expression correlates quantitatively with the tumor cells' capabilities to bind to DPP IV and to metastasize to the lungs. DPP IV/FN mediated adhesion and metastasis are blocked when tumor cells are incubated with soluble DPP IV prior to conducting adhesion and lung colony assays. Adhesion is also blocked by anti-DPP IV monoclonal antibody 6A3 and anti-FN antiserum. However, adhesion to immobilized FN is unaffected by soluble plasma FN and, thus, can happen during hematogenous spread of cancer cells at high plasma FN concentrations. The ability of many cancer cells to capture FN molecules on their surface and to augment such deposits by FN self-association during passage in the blood suggests that DPP IV/FN binding may be a relatively common mechanism for lung metastasis. PMID- 9727045 TI - Multiple receptor domains interact to permit, or restrict, androgen-specific gene activation. AB - A critical problem within transcription factor families is how diverse regulatory programs are directed by highly related members. Androgen and glucocorticoid receptors (AR, GR) recognize a consensus DNA hormone response element (HRE), but they activate target genes with precise specificity, largely dependent on the promoter and cell context. We have assessed the role of different receptor domains in hormone-specific response by testing chimeras of AR and GR for their ability to activate the androgen-specific enhancer of the mouse sex-limited protein (Slp) gene. Although all of the mutant receptors activated simple HREs, only a few activated the androgen-specific element. One component shared by receptors functional on the AR-specific target was the AR DNA binding domain. Activation was not due to differential DNA affinity but rather to the AR DNA binding domain escaping suppression directed at the GR DNA binding domain in this enhancer context. A further mechanism increasing specific activation was cooperation of receptors at multiple and weak HREs, which was accentuated in the presence of both the AR N terminus and ligand binding domain. These domains together increased recognition of weak HREs, as demonstrated by in vitro DNase I footprinting and transactivation of mutant enhancers. Further, AR N-terminal subdomains reported to interact directly with the ligand binding domain relieved an inhibitory effect imposed by that domain. Therefore, functions intrinsic to AR augment steroid-specific gene activation, by evading negative regulation operating on the domains of other receptors and by enhancing cooperativity through intra- and inter-receptor domain interactions. These subtle distinctions in AR and GR behavior enforce transcriptional specificity established by the context of nonreceptor factors. PMID- 9727046 TI - Identity of the beta-globin locus control region binding protein HS2NF5 as the mammalian homolog of the notch-regulated transcription factor suppressor of hairless. AB - Previously, we characterized a DNA-binding protein, HS2NF5, that bound tightly to a conserved region within hypersensitive site 2 (HS2) of the human beta-globin locus control region (LCR) (Lam, L. T. , and Bresnick, E. H. (1996) J. Biol. Chem. 271, 32421-32429). The beta-globin LCR controls the chromatin structure, transcription, and replication of the beta-globin genes. We have now purified HS2NF5 to near-homogeneity from fetal bovine thymus. Two polypeptides of 56 and 61 kDa copurified with the DNA binding activity. The two proteins bound to the LCR recognition site with an affinity (3.1 nM) and specificity similar to mouse erythroleukemia cell HS2NF5. The amino acid sequences of tryptic peptides of purified HS2NF5 revealed it to be identical to the murine homolog of the suppressor of hairless transcription factor, also known as recombination signal binding protein Jkappa or C promoter binding factor 1 (CBF1). The CBF1 site within HS2 resides near sites for hematopoietic regulators such as GATA-1, NF-E2, and TAL1. An additional conserved, high affinity CBF1 site was localized within HS4 of the LCR. As CBF1 is a downstream target of the Notch signaling pathway, we propose that Notch may modulate LCR activity during hematopoiesis. PMID- 9727048 TI - Transcriptional regulatory patterns of the myelin basic protein and malic enzyme genes by the thyroid hormone receptors alpha1 and beta1. AB - While there is evidence that the two ubiquitously expressed thyroid hormone (T3) receptors, TRalpha1 and TRbeta1, have distinct functional specificities, the mechanism by which they discriminate potential target genes remains largely unexplained. In this study, we demonstrate that the thyroid hormone response elements (TRE) from the malic enzyme and myelin basic protein genes (METRE and MBPTRE) respectively, are not functionally equivalent. The METRE, which is a direct repeat motif with a 4-base pair gap between the two half-site hexamers binds thyroid hormone receptor as a heterodimer with 9-cis-retinoic acid receptor (RXR) and mediates a high T3-dependent activation in response to TRalpha1 or TRbeta1 in NIH3T3 cells. In contrast, the MBPTRE, which consists of an inverted palindrome formed by two hexamers spaced by 6 base pairs, confers an efficient transactivation by TRbeta1 but a poor transactivation by TRalpha1. While both receptors form heterodimers with RXR on MBPTRE, the poor transactivation by TRalpha1 correlates also with its ability to bind efficiently as a monomer. This monomer, which is only observed with TRalpha1 bound to MBPTRE, interacts neither with N-CoR nor with SRC-1, explaining its functional inefficacy. However, in Xenopus oocytes, in which RXR proteins are not detectable, the transactivation mediated by TRalpha1 and TRbeta1 is equivalent and independent of a RXR supply, raising the question of the identity of the thyroid hormone receptor partner in these cells. Thus, in mammalian cells, the binding characteristics of TRalpha1 to MBPTRE (i.e. high monomer binding efficiency and low transactivation activity) might explain the particular pattern of T3 responsiveness of MBP gene expression during central nervous system development. PMID- 9727047 TI - The phosphorylation state of CD3gamma influences T cell responsiveness and controls T cell receptor cycling. AB - The T cell receptor (TCR) is internalized following activation of protein kinase C (PKC) via a leucine (Leu)-based motif in CD3gamma. Some studies have indicated that the TCR is recycled back to the cell surface following PKC-mediated internalization. The functional state of recycled TCR and the mechanisms involved in the sorting events following PKC-induced internalization are not known. In this study, we demonstrated that following PKC-induced internalization, the TCR is recycled back to the cell surface in a functional state. TCR recycling was dependent on dephosphorylation of CD3gamma, probably mediated by the serine/threonine protein phosphatase-2A, but independent on microtubules or actin polymerization. Furthermore, in contrast to ligand-mediated TCR sorting, recycling of the TCR was independent of the tyrosine phosphatase CD45 and the Src tyrosine kinases p56(Lck) and p59(Fyn). Studies of mutated TCR and chimeric CD4 CD3gamma molecules demonstrated that CD3gamma did not contain a recycling signal in itself. In contrast, the only sorting information in CD3gamma was the Leu based motif that mediated lysosomal sorting of chimeric CD4-CD3gamma molecules. Finally, we found a correlation between the phosphorylation state of CD3gamma and T cell responsiveness. Based on these observations a physiological role of CD3gamma and TCR cycling is proposed. PMID- 9727049 TI - A p56lck-independent pathway of CD2 signaling involves Jun kinase. AB - The p56 Src family non-receptor tyrosine kinase has been shown to be critical for T lymphocyte differentiation and activation. Hence in the absence of p56, T cell receptor triggered activation does not occur. We now provide evidence for a CD2 based signaling pathway which, in contrast to that of the T cell receptor, is independent of p56. CD2-mediated interleukin-2 production occurs via activation of Jun kinase in cell lines lacking p56. Jun kinase then facilitates the binding of c-Jun/c-Fos heterodimers to the AP-1 consensus site and the subsequent transcriptional activity of the interleukin-2 promoter. These data elucidate differences between TCR and CD2 signaling pathways in the same T cells. PMID- 9727050 TI - Analysis of higher order intermediates and synapsis in the bent-L pathway of bacteriophage lambda site-specific recombination. AB - The integrase protein of bacteriophage lambda mediates recombination via four distinct pathways. The recent in vitro reconstitution of the efficient bidirectional reaction between two attLtenP'1 target sites now allows comparisons of this pathway, known as the bent-L pathway, with the inefficient bidirectional straight-L pathway and with the efficient but unidirectional pathways of integration and excision. To this end, a series of higher order intermediates of the bent-L pathway was characterized using gel mobility shift assays, two dimensional gel analysis, and footprinting. The analysis spans the initial binding of proteins to individual DNA target sites, synapsis of two partner DNA targets, and strand exchange. This study identifies a presynaptic "checkpoint" of recombination. It shows that synapsis is a slow step in the recombination reaction, while subsequent strand exchange is comparatively fast. Synaptic complexes contain a preponderance of recombinant products, suggesting that an energetically favorable but somewhat subtle conformational change drives strand exchange. In addition, comparison of wild-type integrase with a catalytically defective mutant of integrase, IntF, showed that, in addition to the catalysis defect, this mutant has different DNA-binding properties than the wild-type protein. PMID- 9727051 TI - Post-transcriptional regulation of endothelial nitric oxide synthase mRNA stability by Rho GTPase. AB - The mechanism by which 3-hydroxy-3-methylglutaryl (HMG)-CoA reductase inhibitors increase endothelial nitric oxide synthase (eNOS) expression is unknown. To determine whether changes in isoprenoid synthesis affects eNOS expression, human endothelial cells were treated with the HMG-CoA reductase inhibitor, mevastatin (1-10 microM), in the presence of L-mevalonate (200 microM), geranylgeranylpyrophosphate (GGPP, 1-10 microM), farnesylpyrophosphate (FPP, 5-10 microM), or low density lipoprotein (LDL, 1 mg/ml). Mevastatin increased eNOS mRNA and protein levels by 305 +/- 15% and 180 +/- 11%, respectively. Co treatment with L-mevalonate or GGPP, but not FPP or LDL, reversed mevastatin's effects. Because Rho GTPases undergo geranylgeranyl modification, we investigated whether Rho regulates eNOS expression. Immunoblot analyses and [35S]GTPgammaS binding assays revealed that mevastatin inhibited Rho membrane translocation and GTP binding activity by 60 +/- 5% and 78 +/- 6%, both of which were reversed by co-treatment with GGPP but not FPP. Furthermore, inhibition of Rho by Clostridium botulinum C3 transferase (50 microg/ml) or by overexpression of a dominant negative N19RhoA mutant increased eNOS expression. In contrast, activation of Rho by Escherichia coli cytotoxic necrotizing factor-1 (200 ng/ml) decreased eNOS expression. These findings indicate that Rho negatively regulates eNOS expression and that HMG-CoA reductase inhibitors up-regulate eNOS expression by blocking Rho geranylgeranylation, which is necessary for its membrane-associated activity. PMID- 9727052 TI - Secretory interleukin-1 receptor antagonist gene expression requires both a PU.1 and a novel composite NF-kappaB/PU.1/ GA-binding protein binding site. AB - The human secretory interleukin-1 receptor antagonist (secretory IL-1Ra) gene is controlled through three lipopolysaccharide (LPS)-responsive promoter elements, one of which was identified as an NF-kappaB binding site. Sequence analysis of the secretory IL-1Ra promoter identified a potential PU.1 binding site located between positions -80 and -90 on the complementary strand overlapping the NF kappaB site. Gel shift analysis using this potential binding site with nuclear extracts from RAW 264.7 macrophages demonstrated the formation of three complexes, one LPS-inducible and two constitutive. The inducible factor was identified as NF-kappaB, and the constitutive factors were identified as PU.1 and GA-binding protein. Site-directed mutagenesis of the -93 to -79 promoter region demonstrated that mutation of either the NF-kappaB 5'-half site or the PU.1/GA binding protein half-site alone did not significantly decrease LPS responsiveness. However, a mutation that disrupted the binding of all three factors resulted in a 50% decrease in LPS responsiveness. A second PU.1 binding site centered at -230 was identified by gel shift and supershift assays. Mutation of the core GGAA region resulted in a 50% decrease in LPS-responsive promoter activity. Mutation of both the distal and proximal LPS response elements led to an almost complete loss of responsiveness. These data therefore suggest that the regulation of IL-1Ra gene expression is a complex event involving the interactions of three different transcription factors with a single cis-acting element and that the two PU.1 binding sites are the major response elements for LPS-induced IL-1Ra gene expression. PMID- 9727053 TI - Towards a rational therapy for hepatitis B. PMID- 9727054 TI - Leptin inhibits insulin secretion by activation of phosphodiesterase 3B. AB - The molecular signaling events by which leptin exerts its functions in vivo are not well delineated. Here, we show a novel leptin signaling mechanism that requires phosphoinositide 3-kinase (PI 3-kinase)-dependent activation of cyclic nucleotide phosphodiesterase 3B (PDE3B) and subsequent suppression of cAMP levels. In pancreatic beta cells, leptin causes the activation of PDE3B, which leads to marked inhibition of glucagon-like peptide-1-stimulated insulin secretion. The effect of leptin is abolished when insulin secretion is induced with cAMP analogues that cannot be hydrolyzed by PDE3B. Selective inhibitors of PDE3B and PI 3-kinase completely prevent the leptin effect on insulin secretion and cAMP accumulation. The results demonstrate that one of the physiological effects of leptin, suppression of insulin secretion, is mediated through activation of PDE3B and suggest PDE3B as a mediator of leptin action in other tissues. PMID- 9727055 TI - Human beta-defensin 2 is a salt-sensitive peptide antibiotic expressed in human lung. AB - Previous studies have implicated the novel peptide antibiotic human beta-defensin 1 (hBD-1) in the pathogenesis of cystic fibrosis. We describe in this report the isolation and characterization of the second member of this defensin family, human beta-defensin 2 (hBD-2). A cDNA for hBD-2 was identified by homology to hBD 1. hBD-2 is expressed diffusely throughout epithelia of many organs, including the lung, where it is found in the surface epithelia and serous cells of the submucosal glands. A specific antibody made of recombinant peptide detected hBD-2 in airway surface fluid of human lung. The fully processed peptide has broad antibacterial activity against many organisms, which is salt sensitive and synergistic with lysozyme and lactoferrin. These data suggest the existence of a family of beta-defensin molecules on mucosal surfaces that in the aggregate contributes to normal host defense. PMID- 9727056 TI - PYK2 in osteoclasts is an adhesion kinase, localized in the sealing zone, activated by ligation of alpha(v)beta3 integrin, and phosphorylated by src kinase. AB - Osteoclast activation is initiated by adhesion to the bone surface, followed by cytoskeletal rearrangement, the formation of the sealing zone, and a polarized ruffled membrane. This study shows that PYK2/CAKbeta/RAFTK, a cytoplasmic kinase related to the focal adhesion kinase, is highly expressed in rat osteoclasts in vivo. Using murine osteoclast-like cells (OCLs) or their mononuclear precursors (pOCs), generated in a coculture of bone marrow and osteoblastic MB1.8 cells, we show: (a) tyrosine phosphorylation of PYK2 upon ligation of beta3 integrins or adhesion of pOCs to serum, vitronectin, osteopontin, or fibronectin but not to laminin or collagen; (b) coimmunoprecipitation of PYK2 and c-Src from OCLs; (c) PYK2 binding to the SH2 domains of Src; (d) marked reduction in tyrosine phosphorylation and kinase activity of PYK2 in OCLs derived from Src (-/-) mice, which do not form actin rings and do not resorb bone; (e) PYK2 phosphorylation by exogeneous c-Src; (f) translocation of PYK2 to the Triton X-100 insoluble cytoskeletal fraction upon adhesion; (g) localization of PYK2 in podosomes and the ring-like structures in OCLs plated on glass and in the sealing zone in OCLs plated on bone; and (h) activation of PYK2, in the presence of MB1.8 cells, parallels the formation of sealing zones and pit resorption in vitro and is reduced by echistatin or calcitonin and cytochalasin D. Taken together, these findings suggest that Src-dependent tyrosine phosphorylation of PYK2 is involved in the adhesion-induced formation of the sealing zone, required for osteoclastic bone resorption. PMID- 9727057 TI - Lipoprotein lipase expression exclusively in liver. A mouse model for metabolism in the neonatal period and during cachexia. AB - Lipoprotein lipase (LPL), the rate-limiting enzyme in triglyceride hydrolysis, is normally not expressed in the liver of adult humans and animals. However, liver LPL is found in the perinatal period, and in adults it can be induced by cytokines. To study the metabolic consequences of liver LPL expression, transgenic mice producing human LPL specifically in the liver were generated and crossed onto the LPL knockout (LPL0) background. LPL expression exclusively in liver rescued LPL0 mice from neonatal death. The mice developed a severe cachexia during high fat suckling, but caught up in weight after switching to a chow diet. At 18 h of age, compared with LPL0 mice, liver-only LPL-expressing mice had equally elevated triglycerides (10,700 vs. 14,800 mg/dl, P = NS), increased plasma ketones (4.3 vs. 1.7 mg/dl, P < 0.05) and glucose (28 vs. 15 mg/dl, P < 0.05), and excessive amounts of intracellular liver lipid droplets. Adult mice expressing LPL exclusively in liver had slower VLDL turnover than wild-type mice, but greater VLDL mass clearance, increased VLDL triglyceride production, and three- to fourfold more plasma ketones. In summary, it appears that liver LPL shunts circulating triglycerides to the liver, which results in a futile cycle of enhanced VLDL production and increased ketone production, and subsequently spares glucose. This may be important to sustain brain and muscle function at times of metabolic stress with limited glucose availability. PMID- 9727058 TI - beta2-glycoprotein-I (apolipoprotein H) and beta2-glycoprotein-I-phospholipid complex harbor a recognition site for the endocytic receptor megalin. AB - Screening of serum by using a surface plasmon resonance analysis assay identified beta2-glycoprotein-I/apolipoprotein H as a plasma component binding to the renal epithelial endocytic receptor megalin. A calcium-dependent megalin-mediated beta2 glycoprotein-I endocytosis was subsequently demonstrated by ligand blotting of rabbit renal cortex and uptake analysis in megalin-expressing cells. Immunohistochemical and immunoelectron microscopic examination of kidneys and the presence of high concentrations of beta2-glycoprotein-I in urine of mice with disrupted megalin gene established that megalin is the renal clearance receptor for beta2-glycoprotein-I. A significant increase in functional affinity for purified megalin was observed when beta2-glycoprotein-I was bound to the acidic phospholipids, phosphatidylserine and cardiolipin. The binding of beta2 glycoprotein-I and beta2-glycoprotein-I- phospholipid complexes to megalin was completely blocked by receptor-associated protein. In conclusion, we have demonstrated a novel receptor recognition feature of beta2-glycoprotein-I. In addition to explaining the high urinary excretion of beta2-glycoprotein-I in patients with renal tubule failure, the data provide molecular evidence for the suggested function of beta2-glycoprotein-I as a linking molecule mediating cellular recognition of phosphatidylserine-exposing particles. PMID- 9727059 TI - Immunoglobulin treatment reduces atherosclerosis in apo E knockout mice. AB - Atherosclerosis is associated with immune activation. T cells and macrophages infiltrate atherosclerotic plaques and disease progression is associated with formation of autoantibodies to oxidized lipoproteins. In the apo E knockout mouse, a genetic model of cholesterol-induced atherosclerosis, congenital deficiency of macrophages, lymphocytes, or interferon-gamma receptors result in reduced lesion formation. We have now evaluated whether immune modulation in the adult animal affects disease development. Injections of 7-wk-old male apo E knockout mice with polyclonal immunoglobulin preparations (ivIg) during a 5-d period reduced fatty streak formation over a 2-mo period on cholesterol diet by 35%. Fibrofatty lesions induced by diet treatment for 4 mo were reduced by 50% in mice receiving ivIg after 2 mo on the diet. ivIg treatment also reduced IgM antibodies to oxidized LDL and led to inactivation of spleen and lymph node T cells. These data indicate that ivIg inhibits atherosclerosis, that it is effective both during the fatty streak and plaque phases, and that it may act by modulating T cell activity and/or antibody production. Therefore, immunomodulation may be an effective way to prevent and/or treat atherosclerosis. PMID- 9727060 TI - Coordinated induction of plasminogen activator inhibitor-1 (PAI-1) and inhibition of plasminogen activator gene expression by hypoxia promotes pulmonary vascular fibrin deposition. AB - Oxygen deprivation, as occurs during tissue ischemia, tips the natural anticoagulant/procoagulant balance of the endovascular wall to favor activation of coagulation. To investigate the effects of low ambient oxygen tension on the fibrinolytic system, mice were placed in a hypoxic environment with pO2 < 40 Torr. Plasma levels of plasminogen activator inhibitor-1 (PAI-1) antigen, detected by ELISA, increased in a time-dependent fashion after hypoxic exposure (increased as early as 4 h, P < 0.05 vs. normoxic controls), and were accompanied by an increase in plasma PAI-1 activity by 4 h (P < 0.05 vs. normoxic controls). Northern analysis of hypoxic murine lung demonstrated an increase in PAI-1 mRNA compared with normoxic controls; in contrast, transcripts for both tissue-type plasminogen activator (tPA) and urokinase-type plasminogen activator (uPA) decreased under hypoxic conditions. Immunocolocalization studies identified macrophages as the predominant source of increased PAI-1 within hypoxic lung. Using a transformed murine macrophage line, striking induction of PAI-1 transcripts occurred under hypoxic conditions, due to both increased de novo transcription as well as increased mRNA stability. Consistent with an important role of the fibrinolytic system in hypoxia-induced fibrin accumulation, PAI-1 +/+ mice exposed to hypoxia exhibited increased pulmonary fibrin deposition based upon a fibrin immunoblot, intravascular fibrin identified by immunostaining, and increased accumulation of 125I-fibrinogen/fibrin in hypoxic tissue. In contrast, mice deficient for the PAI-1 gene (PAI-1 -/-) similarly exposed to hypoxic conditions did not display increased fibrin accumulation compared with normoxic PAI-1 +/+ controls. Furthermore, homozygous null uPA (uPA -/-) and tPA (tPA -/-) mice subjected to oxygen deprivation showed increased fibrin deposition compared with wild-type controls. These studies identify enhanced expression of PAI-1 as an important mechanism suppressing fibrinolysis under conditions of low oxygen tension, a response which may be further amplified by decreased expression of plasminogen activators. Taken together, these data provide insight into an important potential role of macrophages and the fibrinolytic system in ischemia induced thrombosis. PMID- 9727061 TI - A requirement for the rac1 GTPase in the signal transduction pathway leading to cardiac myocyte hypertrophy. AB - We have used adenoviral-mediated gene transfer of a constitutively active (V12rac1) and dominant negative (N17rac1) isoform of rac1 to assess the role of this small GTPase in cardiac myocyte hypertrophy. Expression of V12rac1 in neonatal cardiac myocytes results in sarcomeric reorganization and an increase in cell size that is indistinguishable from ligand-stimulated hypertrophy. In addition, V12rac1 expression leads to an increase in atrial natriuretic peptide secretion. In contrast, expression of N17rac1, but not a truncated form of Raf-1, attenuated the morphological hypertrophy associated with phenylephrine stimulation. Consistent with the observed effects on morphology, expression of V12rac1 resulted in an increase in new protein synthesis, while N17rac1 expression inhibited phenylephrine-induced leucine incorporation. These results suggest rac1 is an essential element of the signaling pathway leading to cardiac myocyte hypertrophy. PMID- 9727062 TI - Antigen-driven clonal proliferation of B cells within the target tissue of an autoimmune disease. The salivary glands of patients with Sjogren's syndrome. AB - Structures resembling germinal centers are seen in the salivary glands of patients with Sjogren's syndrome, but it is not known whether the microenvironment of these cell clusters is sufficient for the induction of a germinal center response. Therefore, we cloned and sequenced rearranged Ig V genes expressed by B cells isolated from sections of labial salivary gland biopsies from two Sjogren's syndrome patients. Rearranged V genes from B cells within one cell cluster were polyclonal and most had few somatic mutations. Two adjacent clusters from another patient each contained one dominant B cell clone expressing hypermutated V genes. None of the rearranged V genes was found in both clusters, suggesting that cells are unable to migrate out into the surrounding tissue and seed new clusters. The ratios of replacement to silent mutations in the framework and complementarity determining regions suggest antigen selection of high-affinity mutants. These results show that an antigen-driven, germinal center-type B cell response is taking place within the salivary glands of Sjogren's syndrome patients. In view of the recent demonstration of a germinal center response within the rheumatoid synovial membrane and the existence of similar structures in the target tissues of other autoimmune diseases, we propose that germinal center- type responses can be induced in the nonlymphoid target tissues of a variety of autoimmune diseases. PMID- 9727063 TI - Induction of insulitis by glutamic acid decarboxylase peptide-specific and HLA DQ8-restricted CD4(+) T cells from human DQ transgenic mice. AB - Insulin-dependent diabetes mellitus in humans is linked with specific HLA class II genes, e.g., HLA-DQA1*0301/ DQB1*0302 (DQ8). To investigate the roles of HLA DQ8 molecules and glutamic acid decarboxylase (GAD) in disease development, we generated DQ8(+)/I-Abo transgenic mice expressing functional HLA-DQ8 molecules and devoid of endogenous mouse class II. DQ8(+)/I-Abo mice produced antigen specific antibodies and formed germinal centers after immunization with GAD65 peptides. Two GAD peptide-specific (247-266 and 509-528), DQ8 restricted Th1 CD4(+) T cell lines, were generated from immunized DQ8(+)/I-Abo mice. They induced severe insulitis after adoptive transfer into transgene positive (but not negative) mice who were treated with a very low dose of streptozotocin that alone caused no apparent islet pathology. In addition to CD4, islet mRNA from these mice also showed expression of CD8, IFNgamma, TNFalpha, Fas, and Fas ligand. Our data suggest that a mild islet insult in the presence of HLA-DQ8 bearing antigen presenting cells promotes infiltration of GAD peptide reactive T cells into the islet. PMID- 9727064 TI - Developmental upregulation of human parathyroid hormone (PTH)/PTH-related peptide receptor gene expression from conserved and human-specific promoters. AB - The parathyroid hormone (PTH)/PTH-related peptide (PTHrP) receptor (PTHR) functions in skeletal development and mediates an array of other physiological responses modulated by PTH and PTHrP. PTHR gene transcription in mouse is controlled by two promoters: P1, which is highly and selectively active in kidney; and P2, which functions in a variety of tissues. P1 and P2 are conserved in human tissue; however, P1 activity in kidney is weak. We have now identified a third human promoter, P3, which is widely expressed and accounts for approximately 80% of renal PTHR transcripts in the adult. No P3 activity was detected in mouse kidney, indicating that renal PTHR gene expression is controlled by different signals in human and mouse. During development, only P2 is active at midgestation in many human tissues, including calvaria and long bone. This strongly suggests that factors regulating well conserved P2 control PTHR gene expression during skeletal development. Our results indicate that human PTHR gene transcription is upregulated late in development with the induction of both P1 and P3 promoter activities. In addition, P2-specific transcripts are differentially spliced in a number of human cell lines and adult tissues, but not in fetal tissues, giving rise to a shorter and less structured 5' UTR. Thus, our studies show that both human PTHR gene transcription and mRNA splicing are developmentally regulated. Moreover, our data indicate that renal and nonrenal PTHR gene expression are tightly coordinated in humans. PMID- 9727065 TI - Lamivudine treatment can restore T cell responsiveness in chronic hepatitis B. AB - High viral and/or antigen load may be an important cause of the T cell hyporesponsiveness to hepatitis B virus (HBV) antigens that is often observed in patients with chronic HBV infection. Reduction of viral and antigen load by lamivudine treatment represents an ideal model for investigating this hypothesis. HLA class II restricted T cell responses and serum levels of HBV-DNA, HBsAg, and HBeAg were studied before and during lamivudine treatment in 12 patients with hepatitis B e antigen positive chronic active hepatitis B to assess possible correlations between viral and/or antigen load and vigor of the T cell response. Cell proliferation to HBV nucleocapsid antigens and peptides and frequency of circulating HBV nucleocapsid-specific T cells were assessed to characterize CD4 mediated responses. A highly significant enhancement of the CD4-mediated response to HBV nucleocapsid antigens was already detectable in most patients 7-14 d after the start of lamivudine treatment. This effect was dramatic and persistent in 10 patients but undetectable in 2. It occurred concomitant with a rapid and marked reduction of viremia. Interestingly, lamivudine also enhanced the responses to mitogens and recall antigens, showing that its effect was not limited to HBV specific T cells. In conclusion, an efficient antiviral T cell response can be restored by lamivudine treatment in patients with chronic hepatitis B concurrently with reduction of viremia, indicating the importance of viral load in the pathogenesis of T cell hyporesponsiveness in these patients. Since lamivudine treatment can overcome T cell hyporeactivity, combining lamivudine with treatments directed to stimulate the T cell response may represent an effective strategy to induce eradication of chronic HBV infection. PMID- 9727066 TI - An in vitro model for cytogenetic conversion in CML. Interferon-alpha preferentially inhibits the outgrowth of malignant stem cells preserved in long term culture. AB - IFN-alpha has been shown to prolong survival in chronic myeloid leukemia patients, but its mechanism of action is still not understood. The human cobblestone area-forming cell (CAFC) assay allows for the measurement of the concentration of normal as well as malignant stem cells, while their progeny can be measured in parallel long-term culture (LTC) in flasks. Using CAFC and LTC assays, we have examined direct effects of IFN-alpha (500; 5,000 IU/ml) on the maintenance and outgrowth of CD34-enriched normal and malignant stem cells, obtained from six patients with an established major cytogenetic response to IFN alpha and from four nonresponding patients. CAFC concentrations were not affected by IFN-alpha. In contrast, IFN-alpha strongly inhibited the clonogenic output in flask LTC. Nucleated cells (NC) produced in LTC were evaluated by fluorescent in situ hybridization (FISH) for the presence of the Philadelphia (Ph) translocation. After 8 wk of LTC, the percentage of Ph+ NCs produced was significantly more inhibited by IFN-alpha in responding patients than in nonresponders. Control LTC without IFN-alpha showed no significant differences of Ph+ NC production between responders and nonresponders. These findings provide the first in vitro model for cytogenetic conversion and suggest that direct antiproliferative effects of IFN-alpha account for the cytogenetic response observed clinically. PMID- 9727067 TI - Vbeta4(+) T cells promote clearance of infection in murine pulmonary histoplasmosis. AB - T cells are essential for controlling infection with Histoplasma capsulatum. Because the T cell receptor is vital for transducing the biological activities of these cells, we sought to determine if exposure to this fungus induced an alteration in the Vbeta repertoire in lungs of C57BL/6 mice infected intranasally. Vbeta2(+) cells were elevated on day 3 after infection; Vbeta4(+) cells were higher than controls on days 7, 10, and 14 after infection. Vbeta10(+) cells were increased on days 14 and 21, and Vbeta11(+) exceeded controls only on day 14. We investigated the clonality and function of Vbeta4(+) cells because their expansion transpired during the critical time of infection, that is, when cellular immunity is activated. Sequence analysis demonstrated preferential use of a restricted set of sequences in the complementarity-determining region 3. Elimination of Vbeta4(+) cells from mice impaired their ability to resolve infection. In contrast, depletion of Vbeta7(+) cells, the abundance of which was similar to that of Vbeta4(+), did not alter elimination of the fungus. The identification of clonotypes of Vbeta4(+) cells suggests that a few antigenic determinants may drive proliferation of this subset, which is necessary for optimal clearance. PMID- 9727068 TI - CCAAT enhancer- binding protein beta is required for normal hepatocyte proliferation in mice after partial hepatectomy. AB - After two-thirds hepatectomy, normally quiescent liver cells are stimulated to reenter the cell cycle and proliferate to restore the original liver mass. The level of bZIP transcription factor CCAAT enhancer-binding protein beta (C/EBPbeta) increases in the liver during the period of cell proliferation. The significance of this change in C/EBP expression is not understood. To determine the role of C/EBPbeta in the regenerating liver, we examined the regenerative response after partial hepatectomy in mice that contain a targeted disruption of the C/EBPbeta gene. Posthepatectomy, hepatocyte DNA synthesis was decreased to 25% of normal in C/EBPbeta -/- mice. The reduced regenerative response was associated with a prolonged period of hypoglycemia that was independent of expression of C/EBPalpha protein and gluconeogenic genes. C/EBPbeta -/- livers showed reduced expression of immediate-early growth-control genes including the Egr-1 transcription factor, mitogen-activated protein kinase protein tyrosine phosphatase (MKP-1), and HRS, a delayed-early gene that encodes an mRNA splicing protein. Cyclin B and E gene expression were dramatically reduced in C/EBPbeta -/ livers whereas cyclin D1 expression was normal. The abnormalities in immediate early gene expression in C/EBPbeta -/- livers were distinct from those seen in IL 6 -/- livers. These data link C/EBPbeta to the activation of metabolic and growth response pathways in the regenerating liver and demonstrate that C/EBPbeta is required for a normal proliferative response. PMID- 9727069 TI - Active sodium-urea counter-transport is inducible in the basolateral membrane of rat renal initial inner medullary collecting ducts. AB - Rat inner medullary collecting ducts (IMCD3s) possess a luminal Na+-dependent, active urea secretory transport process, which is upregulated by water diuresis. In this study of perfused IMCDs microdissected from base (IMCD1), middle (IMCD2), or tip (IMCD3) of the inner medulla, we tested whether furosemide diuresis alters active urea transport. Rats received furosemide (10 mg/d s.c. for 3-4 d) and were compared with pair-fed control rats. Furosemide significantly decreased urine osmolality and urea clearance, and increased blood urea nitrogen. IMCD3s from furosemide-treated rats had significantly lower rates of active urea secretion than IMCD3s from control rats. IMCD2s showed no active urea transport in control or furosemide-treated rats. IMCD1s from control rats had no active urea transport, but IMCD1s from furosemide-treated rats expressed significant rates of active urea reabsorption. In IMCD1s, this active urea reabsorptive transport process was inhibited by: (i) 0. 25 mM phloretin (bath); (ii) 1 mM ouabain (bath); and (iii) replacing bath Na+ with NMDG+; it was stimulated by 10 nM bumetanide (bath). In summary, we found that furosemide decreased active urea secretion in IMCD3s and induced active urea reabsorption in IMCD1s. The new Na+- dependent, active urea reabsorptive transport process may be a basolateral Na+ urea antiporter. PMID- 9727070 TI - The human orphan nuclear receptor PXR is activated by compounds that regulate CYP3A4 gene expression and cause drug interactions. AB - The cytochrome P-450 monooxygenase 3A4 (CYP3A4) is responsible for the oxidative metabolism of a wide variety of xenobiotics including an estimated 60% of all clinically used drugs. Although expression of the CYP3A4 gene is known to be induced in response to a variety of compounds, the mechanism underlying this induction, which represents a basis for drug interactions in patients, has remained unclear. We report the identification of a human (h) orphan nuclear receptor, termed the pregnane X receptor (PXR), that binds to a response element in the CYP3A4 promoter and is activated by a range of drugs known to induce CYP3A4 expression. Comparison of hPXR with the recently cloned mouse PXR reveals marked differences in their activation by certain drugs, which may account in part for the species-specific effects of compounds on CYP3A gene expression. These findings provide a molecular explanation for the ability of disparate chemicals to induce CYP3A4 levels and, furthermore, provide a basis for developing in vitro assays to aid in predicting whether drugs will interact in humans. PMID- 9727071 TI - Two different functions for CD44 proteins in human myelopoiesis. AB - CD44 is important during myelopoiesis, although the contributions of variant CD44 proteins are unclear. We show here that in human long-term bone marrow culture antibodies recognizing a CD44 NH2-terminal epitope (mab 25-32) or a CD44v6 epitope (mab VFF18) inhibit myelopoiesis. However, mab 25-32 but not mab VFF18 affects myeloid colony formation. These data suggest that an early precursor cell compartment is the target for the 25-32 antibody, whereas the mab VFF18 targets later stages in myelopoiesis. Since the bulk of hemopoietic precursor cells are negative for the v6 epitope and only a minor subset of myeloid cells express the v6 epitope, we have used several human myeloid progenitor cell lines to unravel the function of different CD44 proteins. These cell lines produce variant CD44 proteins, predominantly a new variant CD44v4-v10, when stimulated towards myeloid differentiation. Features that can be acquired by the expression of CD44v4-v10 are an increased hyaluronate (HA) and a de novo chondroitin sulphate A (CS-A) binding. Although, the expression of CD44v4-v10 per se is necessary for HA and CS A binding, the protein backbone seems to require appropriate glycosylation. HA binding results in CD44-mediated cellular self-aggregation and adhesion to the stromal cell line MS-5. In summary, our data suggest that different CD44 proteins are important for at least two different steps in myelopoiesis. PMID- 9727072 TI - The route of estrogen replacement therapy confers divergent effects on substrate oxidation and body composition in postmenopausal women. AB - The route of estrogen replacement therapy has a major impact on the growth hormone (GH)/insulin-like growth factor-I (IGF-I) axis. Estrogen administration by the oral, but not the transdermal route, reduces IGF-I and increases GH levels in postmenopausal women. To investigate whether these perturbations have metabolic consequences, we compared the effects of 24 wk each of oral (Premarin 1.25 mg) and transdermal (Estraderm 100TTS) estrogen on energy metabolism and body composition in 18 postmenopausal women in an open-label randomized crossover study. Energy expenditure, lipid oxidation (lipid(ox)), and carbohydrate oxidation (CHOox) were measured by indirect calorimetry in the fasted and fed state before and after 2 and 6 mon treatment. Lean body mass, fat mass, and total body bone mineral density were measured by dual X-ray absorptiometry before and after 6 mon treatment. Mean (+/-SE) Luteinizing hormone levels fell to comparable levels during oral and transdermal estrogen, and bone mineral density was significantly increased by both treatments. Mean IGF-I was significantly lower during oral estrogen (77+/-7 versus 97+/-7 microg/liter, P < 0.05) treatment. Lipid(ox) 30-60 min after a standardized meal was significantly lower (36+/-5 versus 54+/-5 mg/min, P < 0.01) and CHOox higher (147+/-13 versus 109+/-12 mg/min, P < 0.05) with oral compared with transdermal estrogen. Oral estrogen resulted in a 1.2+/-0.5 kg (P < 0.05) increase in fat mass and a 1.2+/-0.4 kg (P < 0.01) decrease in lean mass compared with transdermal estrogen. Lean body mass (0.4+/-0.2 kg) and fat mass (0. 1+/-0.4 kg) did not change significantly during transdermal estrogen. In summary, when compared with the transdermal route, oral estrogen reduces lipid(ox), increases fat mass, and reduces lean body mass. The route of estrogen therapy confers distinct and divergent effects on substrate oxidation and body composition. The suppression of lipidox during oral estrogen therapy may increase fat mass although the fall in IGF-I may lead to a loss of lean body mass. The route-dependent changes in body composition observed during estrogen replacement therapy may have important implications for postmenopausal health. PMID- 9727073 TI - Mapping a gene involved in regulating dietary cholesterol absorption. The sitosterolemia locus is found at chromosome 2p21. AB - The molecular mechanisms regulating the amount of dietary cholesterol retained in the body as well as the body's ability to selectively exclude other dietary sterols are poorly understood. Studies of the rare autosomal recessively inherited disease sitosterolemia (OMIM 210250) may shed some light on these processes. Patients suffering from this disease appear to hyperabsorb both cholesterol and plant sterols from the intestine. Additionally, there is failure of the liver's ability to preferentially and rapidly excrete these non cholesterol sterols into bile. Consequently, people who suffer from this disease have very elevated plasma plant sterol levels and develop tendon and tuberous xanthomas, accelerated atherosclerosis, and premature coronary artery disease. Identification of this gene defect may therefore throw light on regulation of net dietary cholesterol absorption and lead to an advancement in the management of this important cardiovascular risk factor. By studying 10 well-characterized families with this disorder, we have localized the genetic defect to chromosome 2p21, between microsatellite markers D2S1788 and D2S1352 (maximum lodscore 4.49, theta = 0.0). PMID- 9727074 TI - Clonal expansion and somatic hypermutation of V(H) genes of B cells from cerebrospinal fluid in multiple sclerosis. AB - The cerebrospinal fluid (CSF) of multiple sclerosis (MS) patients is characterized by increased concentrations of immunoglobulin (Ig), which on electrophoretic analysis shows restricted heterogeneity (oligoclonal bands). CSF Ig is composed of both serum and intrathecally produced components. To examine the properties of intrathecal antibody-producing B cells, we analyzed Ig heavy chain variable (V(H)) region genes of B cells recovered from the CSF of 12 MS patients and 15 patients with other neurological diseases (OND). Using a PCR technique, we could detect rearrangements of Ig V(H) genes in all samples. Sequence analysis of complementarity-determining region 3 (CDR3) of rearranged VDJ genes revealed expansion of a dominant clone or clones in 10 of the 12 MS patients. B cell clonal expansion was identified in 3 of 15 OND. The nucleotide sequences of V(H) genes from clonally expanded CSF B cells in MS patients demonstrated the preferential usage of the V(H) IV family. There were numerous somatic mutations, mainly in the CDRs, with a high replacement-to-silent ratio; the mutations were distributed in a way suggesting that these B cells had been positively selected through their antigen receptor. Our results demonstrate that in MS CSF, there is a high frequency of clonally expanded B cells that have properties of postgerminal center memory or antibody-forming lymphocytes. PMID- 9727077 TI - Same Pit, Different Vipers. PMID- 9727075 TI - VLA-5 is expressed by mouse and human long-term repopulating hematopoietic cells and mediates adhesion to extracellular matrix protein fibronectin. AB - Fibronectin (FN), an extracellular matrix protein, is involved in the adhesion and migration of hematopoietic cells and has been shown to enhance retroviral gene transfer into primitive hematopoietic cells by co-localization of target cells and retrovirus when used as a substrate in vitro. We have previously found that mouse hematopoietic stem cells could be transduced on a FN fragment that included the recognition sequence Arg-Gly-Asp (RGD), suggesting that stem cells may express the integrin very late antigen (VLA)-5. To address this, we investigated the binding of mouse and human hematopoietic cells to recombinant peptides that contained one or a combination of the three principle cell-binding domains of FN. These domains included the VLA-5- binding sequence RGD, the VLA-4 binding site CS1, and the high affinity heparin-binding domain. Here we show that mouse long-term in vivo repopulating stem cells, as well as primitive human NOD/SCID mouse repopulating cells, can bind extracellular matrix protein FN by using integrin VLA-5 in vitro. This binding is specific and can be inhibited by antibodies to VLA-5. In addition, preincubation of BM cells with peptide CH-296, which contains all three primary FN-binding domains, decreased the engraftment of cells in the bone marrow in vivo, while intravenous injection of the same peptide induced an increase of progenitor cells in the spleen. In summary, our data demonstrate that VLA-5 is expressed on primitive mouse and human hematopoietic cells and suggest that there may be significant cooperation between integrin receptors and proteoglycan molecules in the engraftment of bone marrow cells and hematopoietic cell adhesion in vivo. PMID- 9727078 TI - Cysteine metabolism and metal toxicity. AB - Chronic, low level exposure to toxic metals is an increasing global problem. The symptoms associated with the slow accumulation of toxic metals are multiple and rather nondescript, and overt expression of toxic effects may not appear until later in life. The sulfhydryl-reactive metals (mercury, cadmium, lead, arsenic) are particularly insidious and can affect a vast array of biochemical and nutritional processes. The primary mechanisms by which the sulfhydryl-reactive metals elicit their toxic effects are summarized. The pro-oxidative effects of the metals are compounded by the fact that the metals also inhibit antioxidative enzymes and deplete intracellular glutathione. The metals also have the potential to disrupt the metabolism and biological activities of many proteins due to their high affinity for free sulfhydryl groups. Cysteine has a pivotal role in inducible, endogenous detoxication mechanisms in the body, and metal exposure taxes cysteine status. The protective effects of glutathione and the metallothioneins are discussed in detail. Basic research pertaining to the transport of toxic metals into the brain is summarized, and a case is made for the use of hydrolyzed whey protein to support metal detoxification and neurological function. Metal exposure also affects essential element status, which can further decrease antioxidation and detoxification processes. Early detection and treatment of metal burden is important for successful detoxification, and optimization of nutritional status is paramount to the prevention and treatment of metal toxicity. PMID- 9727076 TI - Human infection with Trypanosoma cruzi induces parasite antigen-specific cytotoxic T lymphocyte responses. AB - Experimental models of Chagas' disease, an infection caused by the intracellular protozoan Trypanosoma cruzi, have demonstrated the crucial immunoprotective role played by CD8(+) T lymphocytes. These cells dominate inflammatory foci in parasitized tissues and their elimination from mice leads to uncontrolled parasite replication and subsequent death of the infected host. A trypomastigote surface antigen, TSA-1, and two amastigote surface molecules, ASP-1 and ASP-2, were recently identified as targets of CD8(+) cytotoxic T lymphocytes (CTL) in T. cruzi-infected mice. Until now, however, there was no evidence for the development of parasite-specific CTL in T. cruzi-infected humans. In this study, human CTL specific for TSA-1-, ASP-1-, and ASP-2-derived peptides were detected in the peripheral blood mononuclear cells from 21 of 24 HLA-A2(+) T. cruzi infected patients. CTL recognition was antigen specific, A2-restricted, and CD8(+) T cell-dependent. Demonstration of human CTL against T. cruzi and against target molecules identified using the murine model provides important information for the optimal design and evaluation of vaccines to prevent or ameliorate Chagas' disease. PMID- 9727079 TI - Cerebrospinal fluid protein changes in multiple sclerosis after dental amalgam removal. AB - A relationship between multiple sclerosis (MS) and dental silver-mercury fillings has been suggested by some investigators, but never proven. This study documents objective biochemical changes following the removal of these fillings along with other dental materials, utilizing a new health care model of multidisciplinary planning and treatment. The dramatic changes in photolabeling of cerebrospinal fluid (CSF) proteins following these dental interventions suggest CSF photolabeling may serve as an objective biomarker for monitoring MS. The clear cut character of these changes should also encourage more research to better define this possible association between dental mercury and MS. PMID- 9727083 TI - Evolution and phylogeny of defense molecules associated with innate immunity in horseshoe crab. AB - This short review describes the molecular evolution and phylogeny of various defense molecules participating in the host defense of horseshoe crab. It is well known that invertebrate animals, which lack adaptive immune systems, have developed various defense systems, so called innate immunity, that respond to common antigens on the surface of potential pathogens. The systems include hemolymph coagulation, melanization, cell agglutination, antimicrobial action, active oxygen formation, and phagocytic action. Among them, hemolymph coagulation and phenoloxidase-mediated melanization, in addition to cell agglutination, are directly induced by foreign substances, that result in the engulfment of invading microbes. The immobilized invaders are finally killed by antimicrobial substances released mainly from many kinds of hemocytes. In the past two decades, we have investigated biochemically various defense molecules, using horseshoe crab as a model animal, and established extensively their molecular structures. These results now make it possible to discuss evolution and phylogeny of the defense molecules at a molecular level, in comparison with those derived from vertebrate animals. Here, the authors will describe the present state of our knowledge concerning molecules mainly associated with innate immunity. PMID- 9727084 TI - Physiologic importance of protein phosphatase inhibitors. AB - Reversible protein phosphorylation is an important mode of regulation of cellular processes. While earlier studies focused on protein kinases, it is now apparent that protein phosphatases play an equally integral role in the control of cellular phosphoproteins. This review examines the role played by endogenous inhibitors of three major protein serine/threonine phosphatases, PP1, PP2A and PP2B in the control of cell physiology. The discussion highlights novel paradigms for signal transduction by protein phosphatase inhibitors that provide important avenues for signal amplification, the timing of physiological responses and cross talk between distinct signal transduction pathways. New evidence also points to genetic abnormalities or altered expression of phosphatase inhibitors as potential mechanisms for human disease.Together, the data emphasize the physiological importance of protein phosphatase inhibitors and establish phosphatase regulation as a key feature of hormone signaling. PMID- 9727085 TI - Biological and molecular basis of human breast cancer. AB - Human breast cancer remains the most common malignancy in the American women. The ultimate cure of this disease relies on a better understanding of the mechanisms underlying the initiation and progression of this disease. The neoplastic transformation of HBEC in vitro represents a successful model for obtaining knowledge on the molecular and biological alterations that may contribute to the tumorigenic mechanisms. We have presented here a current understanding of chemically transformed HBEC in the following aspects: 1. Factors affecting the transformation of HBEC such as genetic predisposition and differentiation status and prior immortalization; 2. New targets for studying the mechanism of cell immortalization such as alterations in telomerase activity and differential expression of cell cycle dependent genes as well as others recently isolated through differential cloning such as H-ferritin, and a calcium binding protein; 3. Epigenetic and genetic mechanisms underlying cell transformation; 4. The association of microsatellite instability in specific loci on chromosomes 11, 13, and 16 with the progression of cell transformation; and 5. The application of microcell mediated chromosome transfer technique as an approach to testing the functional role of specific genes whose dysregulation or loss of function may contribute to the ultimate cell transformation. Further efforts in this cell system will be directed to determine the roles of identified molecular changes as well as the mapping/cloning of tumor suppressor or senescence genes such as those that may reside on chromosomes 11 or 17. PMID- 9727086 TI - Molecular and genetic analysis of iron uptake proteins in the brazilian purpuric fever clone of Haemophilus influenzae biogroup aegyptius. AB - Haemophilus influenzae biogroup aegyptius (H. aegyptius) is the etiological agent of Brazilian purpuric fever (BPF), a recently described pediatric disease that is often fatal. The vascular destruction that occurs in this disease is a distinctive trait, and little is known about the mechanism(s) of the overwhelming purpura fulminans that causes the high mortality associated with this pediatric infection. Iron is an essential micronutrient for nearly all living cells, and the mechanisms used by bacteria to acquire and internalize iron are often associated with virulence. Therefore, the focus of our studies is the molecular characterization of the iron uptake system used by H. aegyptius. Specifically, we are investigating the high-affinity transferrin binding proteins in the bacterial outer membrane, components of ABC transporter systems, and a possible regulatory mechanism for the genes encoding these proteins. A detailed understanding of the molecular nature of the regulatory genetic components and proteins involved in the acquisition of iron will broaden the knowledge of the pathogenesis of the disease caused by H. aegyptius and will also lead to a better understanding of the nature of other infections that affect the vascular system. PMID- 9727087 TI - Basonuclin, a zinc finger protein associated with epithelial expansion and proliferation. AB - Basonuclin is a zinc finger protein first described as a keratinoycte specific protein present in various stratified squamous epithelia found in epidermis, esophagis, cornea and virgina. Recent studies showed that its distribution also includes the germ cells of testis and ovary. The presence of basonuclin appeared always related to the cellular proliferative ability not just cell division, for it was found both in dividing and quiescent cells. Basonuclin disappeared when a cell became post-mitotic. This review examines the latest findings about the distribution, molecular and cellular biology of basonuclin and discusses its possible role in cell proliferation. PMID- 9727088 TI - 5-Hydroxytryptophan: a clinically-effective serotonin precursor. AB - 5-Hydroxytryptophan (5-HTP) is the intermediate metabolite of the essential amino acid L-tryptophan (LT) in the biosynthesis of serotonin. Intestinal absorption of 5-HTP does not require the presence of a transport molecule, and is not affected by the presence of other amino acids; therefore it may be taken with meals without reducing its effectiveness. Unlike LT, 5-HTP cannot be shunted into niacin or protein production. Therapeutic use of 5-HTP bypasses the conversion of LT into 5-HTP by the enzyme tryptophan hydroxylase, which is the rate-limiting step in the synthesis of serotonin. 5-HTP is well absorbed from an oral dose, with about 70 percent ending up in the bloodstream. It easily crosses the blood brain barrier and effectively increases central nervous system (CNS) synthesis of serotonin. In the CNS, serotonin levels have been implicated in the regulation of sleep, depression, anxiety, aggression, appetite, temperature, sexual behaviour, and pain sensation. Therapeutic administration of 5-HTP has been shown to be effective in treating a wide variety of conditions, including depression, fibromyalgia, binge eating associated with obesity, chronic headaches, and insomnia. PMID- 9727089 TI - Milk and other dietary influences on coronary heart disease. AB - While dietary links to ischemic heart disease (IHD) and coronary heart disease (CHD) mortality have been studied for many years, the correlation has not clearly been resolved, especially for older populations. In this paper, a multi-country statistical approach involving 32 countries is used to find dietary links to IHD and CHD for various age groups aged 35+. For IHD, milk carbohydrates were found to have the highest statistical association for males aged 35+ and females aged 65+, while for females aged 35-64, sugar was found to have the highest association. In the case of CHD, non-fat milk was found to have the highest association for males aged 45+ and females aged 75+, while for females 65-74, milk carbohydrates and sugar had the highest associations, and for females aged 45-64, sugar had the highest association. A number of mechanisms have been proposed in the literature that might explain the milk carbohydrate or non-fat milk association. One of the most prominent theories is that animal proteins contribute to homocysteine (Hcy) production; however, milk more than meat lacks adequate B vitamins to convert Hcy to useful products. Lactose and calcium in conjunction with Hcy from consumption of non-fat milk may also contribute to calcification of the arteries. PMID- 9727090 TI - The correspondence of S.A.S. Simpson and J.F. Tait with T. Reichstein during their collaborative work on the isolation and elucidation of the structure of electrocortin (later aldosterone). PMID- 9727091 TI - Synthesis of (19E)-3 beta,7 alpha-dihydroxy-17-oxoandrost-5-en-19-al 19-(O carboxymethyl)oxime, a new hapten for 7 alpha-hydroxydehydroepiandrosterone (3 beta,7 alpha-dihydroxyandrost-5-en-17-one). AB - The title compound was prepared in 11 steps from 17,17-ethylenedioxy-19 hydroxyandrost-5-en-3 beta-yl acetate. After tert-butyldimethylsilyl protection of the 19-hydroxyl group, a 7-oxo group was introduced by oxidation with 3,5 dimethylpyrazole-chromium trioxide complex, and then selectively reduced with L Selectride to give a 7 alpha-hydroxy derivative. This partially protected triol was acetylated and desilylated to 3,7-diacetate. Subsequent oxidation with pyridine-chromium trioxide complex gave 19-aldehyde, which was transformed into the corresponding protected 19-(O-carboxymethyl)oxime. Successive ketal cleavage, deacetylation, and methyl ester splitting gave the final (19E)-3 beta,7 alpha dihydroxy-17-oxoandrost-5-en-19-al 19-(O-carboxymethyl)oxime, designed as a hapten for 7 alpha-hydroxydehydroepiandrosterone immunoassays. PMID- 9727092 TI - Non-ACTH POMC fragments stimulate aldosterone production by human adrenal cells in vitro. AB - The possibility that non-ACTH proopiomelanocortin-derived fragments may stimulate aldosterone production has previously been studied using nonhuman cells with inconsistent results. We have examined the response of aldosterone to beta endorphin (beta-End) and joining peptide (JP) and compared these with the response to ACTH using eight cell suspensions prepared from human adrenal glands. ACTH, 10(-6), 10(-8), and 10(-10) M, consistently stimulated aldosterone accumulation above that occurring in unstimulated cells (150 +/- 83, 120 +/- 62, and 77 +/- 32 fmol/10(4) cells above basal, respectively; mean +/- SE; p < 0.05). beta-End significantly stimulated aldosterone production at 10(-6) and 10(-8) M (114 +/- 84 and 50 +/- 24 fmol/10(4) cells above basal; p < 0.05); 10(-10) M beta End did not provide significant stimulation. Furthermore, JP stimulated aldosterone biosynthesis (41 +/- 16 fmol/10(4) cells above basal; p < 0.05), only at the highest concentration used, 10(-6) M. The addition of 10(-8) M ACTH plus 10(-6) and 10(-10) M beta-End to human adrenal cells yielded values significantly greater than those achieved with either agent alone (267 +/- 152 and 183 +/- 89 fmol/10(4) cells above basal; p < 0.05). These data indicate for the first time that beta-End and JP have the capacity to stimulate aldosterone production in human adrenal cells in vitro. The physiological and potential clinical significance of these observations has yet to be elucidated. PMID- 9727093 TI - Partial synthetic derivatization of canrenone and characterization of its impact on the inhibitory effect on Na+/K(+)-ATPase activity in human heart muscle. AB - To improve the weak inhibitory effect of 3-oxo-17 alpha-pregna-4,6-diene-21,17 carbolactone (canrenone, II) on Na+/K(+)-ATPase activity in human heart muscle, we have investigated the impact of hydrogenation, reduction, glycosidation, and the introduction of a 3-sulfonamido residue on the inhibitory potency of canrenone. The greatest increase in potency (> 20 times) was found for 3 beta (alpha-L-rhamnopyranosyloxy)-5 beta, 17 alpha-pregnane-21, 17-carbolactone (IX). The 3-O-glycosides IX-XI are the first representatives of C/D-trans steroids with effector-receptor complex decay half-times longer than those of therapeutically used cardenolides. PMID- 9727094 TI - Methyl hypofluorite in the synthesis of 16-methoxyestradiol stereoisomers. AB - The usual chemistry of methyl hypofluorite provides a previously unexplored route for functionalizing the 16-position of estradiol. Three isomers of 16 methoxyestradiol were prepared via two synthetic routes, each using methyl hypofluorite. The estrogen receptor binding affinity of these compounds was determined to evaluate their potential as positron emission tomographic (PET) imaging agents targeting estrogen receptor-positive breast cancer. Radiolabeled methyl hypofluorite ([11C]CH3OF) would allow the rapid preparation of novel carbon-11 PET imaging agents. The 17-trimethylsilyl enol ethers of 3-benzyloxy and 3-trifloxyestrone were prepared as substrates to react with methyl hypofluorite. Conditions for the reaction of methyl hypofluorite with simple substrates were optimized to provide reasonable reaction yields with the steroidal substrates. Following introduction of the methoxy substituent at the 16 position, reduction and deprotection conditions were manipulated to yield the various methoxyestradiol isomers. Two-dimensional NMR techniques (HMQC and HMQC TOCSY) were instrumental in the characterization of the methoxyestradiol isomers. NOESY experiments confirmed the stereochemistry of the 16- and 17-positions. 16 alpha-Methoxyestradiol-17 beta and 16 beta-methoxyestradiol-17 beta each with the preferred beta orientation for the 17-alcohol, were determined to have relative binding affinities of 1.5% and 2.3%, respectively. The stereoisomer with the unfavored alpha orientation at the 17-position, 16 alpha-methoxyestradiol-17 alpha, exhibited only a 0.5% relative binding affinity for the estrogen receptor. The biological evaluation of these compounds was not pursued further because of their low binding affinities. PMID- 9727095 TI - Novel high-affinity steroidal estrogenic ligands: synthesis and receptor binding of 11 beta-vinyl-17 alpha-E/Z-phenylselenovinyl estradiols. AB - Previous studies from our laboratory demonstrated separately the tolerance of the estrogen receptor for the 17 alpha-phenylselenovinyl substituent and the enhancement of affinity imparted by the 11 beta-vinyl moiety. Our recent publication suggested that the two groups could be combined within a single structure and retain high affinity for the estrogen receptor. As a result, we have prepared in good overall yields the E- and Z-isomers of 11 beta-vinyl-17 alpha-phenylselenovinyl estradiol. Evaluation of the new steroids with receptor isolated from lamb cytosol indicated that both isomers are poorer ligands than estradiol at 4 degrees C, but both are better than estradiols. at 25 degrees C. This behavior had not been observed for the 11 beta-unsubstituted 17 alpha-E/Z phenylselenovinyl estradiols. Of particular interest was the observation that, unlike previous isomer pairs, the E-isomer possessed a greater affinity than the Z-isomer. The results suggest that relatively small changes in structure may impart significant differences in the interactions with the receptor and provide the basis for further ligand design. PMID- 9727097 TI - Waiting for a new heart: life on a rope. PMID- 9727096 TI - Microbial hydroxylation of acetylaminosteroids. AB - An investigation of the microbial biotransformation of a range of 3 beta-, 17 beta-, and 20-acetylamino C18 to C21 steroids by microorganisms known to hydroxylate conventional steroids has been undertaken, using Aspergillus ochraceus, Bacillus megaterium, Curvularia lunata, and Rhizoputus arrhizus. A. ochraceus and B. megaterium gave products of 11 alpha- and 15 beta-hydroxylation, respectively. In the case of C. lunata, the products were predominantly those of this organism's normal C-11 beta- and C-14 alpha-hydroxylating pathways, but in one instance, 3 beta-acetylamino-7 alpha-hydroxy-5 alpha-androstan-17-one, appeared to results from direction of the site of hydroxylation by the substitution pattern of the substrate. The products from R. arrhizus generally corresponded to those previously obtained from normal steroids of similar skeleton, with 6 beta- 11 alpha-hydroxylation predominating, but again the sites of hydroxylation and the range of hydroxylated products were found to depend on the substitution pattern of the substrate. PMID- 9727098 TI - Consent for cadaver organ and tissue donation. PMID- 9727099 TI - Hospital development and the performance of organ procurement organizations. AB - With more than 56,000 patients on the national waiting list for organ transplants and relatively little growth in the number of donors, organ procurement organizations now recognize the need to aggressively market their services and the range of donor procurement opportunities to hospital personnel. This study examines the types and levels of hospital development activities being conducted by organ procurement organizations, the characteristics of organ procurement organizations that are more involved in hospital development, and the relationship between hospital development and organ procurement. Results from a national survey indicate that, as of the mid-1990s, organ procurement organizations had not made major investments in hospital development despite an increased recognition of the importance of these activities. Organ procurement organizations whose directors were more committed to hospital development exhibited higher levels of hospital development activity. Efforts to formalize hospital development activities through the establishment of a hospital development department and evaluation standards were associated with more organs procured per donor. PMID- 9727100 TI - An in-house coordinator program to increase organ donation in public trauma hospitals. AB - A 4-year retrospective study was conducted regarding the donor potential, consent rates, and organ recovery at a large 500-bed public trauma hospital. An independent organ procurement organization hired two in-house coordinators, one white and one black, to work exclusively in the hospital. The duties of the in house coordinators included the following: working with nurses, physicians, and residents to identify donors; closely managing and coordinating the consent process; and assisting organ procurement coordinators in donor management. Following the program's implementation and the use of race-specific requesters, a 64% increase in consent rate resulted along with an overall increase of 94% in the number of organ donors. The consent rate of blacks increase 115%, whereas the number of black organ donors increased 154%. The Hispanic consent rate increased 48% with a corresponding increase of 83% in the number of Hispanic organ donors. In addition, the white consent rate increased from 55% (the 3-year average from 1993 to 1995) to 75% in 1996, resulting in a 36% increase following the implementation of the program. The investment of dedicated race-sensitive personnel in large urban county trauma facilities can result in a significant increase in donor conversion rates. PMID- 9727101 TI - The coordinator attrition problem in the United States: myth or reality? AB - Excessive attrition among organ procurement coordinators has been suspected problem for many years. In 1994 the United Network for Organ Sharing undertook a procurement coordinator attrition study. Initially, focus groups were conducted in conjunction with the 1994 North American Transplant Coordinators Organization's annual meeting. In 1996, 69 executive directors from organ procurement organizations were surveyed on the subject of procurement coordinator attrition. Thirty-five executive directors responded, resulting in a 51% response rate. The UNOS procurement coordinator attrition study explored actual attrition rates, relationships of certain demographic factors to attrition rates, economic impact of attrition on the organizations, and various job tenure issues. The period under study was January 1, 1990 through December 31, 1993. Results did not demonstrate an industry-wide attrition problem. Additionally, the study revealed no enduring attrition problem in any single organization, and some organizations were found to have no attrition during the entire study period. PMID- 9727102 TI - Standardizing donation education for nursing students: one organ procurement organization's approach. AB - Nurses are often the primary source of donor referrals to an organ procurement organization. Early exposure to donation information during nursing school may enhance nurses' ability to identify and refer potential donors once they are in nursing practice. Although organ procurement organizations strive to provide donation education to nursing students, this process can drain an organization's resources. Therefore, to facilitate the educational process with a minimum of disruption to the function of the organ procurement organization, a standardized approach to donation was developed. After the program had been in place for more than a year, students and organ procurement organization staff alike reported great satisfaction with this standardized approach. PMID- 9727103 TI - Adequate consent: its content in the donation discussion. AB - Although recent research has established a relationship between donation rates and the consent process for organ and tissue donation, little attention has been given to the content of the donation discussion. Recent studies suggest that families want more information to aid them in making the decision to consent. To address the family's needs, the interviewer should introduce the subject appropriately, respond fully to the family's questions, and provide the following information: the options of donating particular organs or tissues, the potential benefits of donation, the right not to donate, the effects on the funeral, the lack of cost of donation to the family, a description of the procurement procedures, and what will happen if the donated organs cannot be used for transplant. PMID- 9727104 TI - Incidence and treatment of cancer in transplant recipients. AB - Solid organ transplantation has been an accepted mode of therapy for the treatment of end-stage organ diseases for many years. Recipients' survival, however, has been hindered by organ rejection and the comorbid diseases that develop as a result of immunosuppresive therapy. In particular, organ transplant recipients have an increased risk of developing cancer de novo after transplantation. Prevalence ranges from 4 to 18% with an average incidence of 6%. Data submitted to the Cincinnati transplant tumor registry have revealed that cancers prevalent in the general population exhibited no increase in rate and may even show a slight decline in the transplant population. Length of survival after transplantation is associated with the likelihood of having cancer; the longer the recipient survives, the greater the chance. The actuarial risk among 124 cardiac transplant recipients was 2.7 +/- 1.9% at 1 year and 25.6 +/- 11% at 5 years. This article will review the current literature on the incidence and treatment of cancer in solid organ transplant recipients. PMID- 9727105 TI - Assessment of quality of life and related stressors following liver transplantation. AB - The aim of this descriptive study was to identify and compare transplant associated stressors among 30 adult liver transplant recipients following orthotopic transplantation for primary biliary cirrhosis. Recipients were divided in three groups according to length of time following transplantation: 12 months (group A), 1 to 5 years (group B), and 5 or more years (group C). With the use of a recipient stressor scale and a rating scale assessing health-related quality of live, patients were asked to indicate a degree of concern for 25 items. In addition, participants were asked to define quality of life before and after liver transplantation and to attribute a degree of positivity or negativity to 33 items in the subscales of physical health, physical activity, social activity, and general effect. Results from the stressor analysis identified health-related items, in particular the "possibility of rejection," as causing the greatest concern to respondents. Social and family-related items were considered least stressful. Comparative statistical analysis identified group B as significantly more stressed than either group A or group C. PMID- 9727106 TI - A comparison of role expectations and communication styles between transplant coordinators and transplant staff nurses. AB - The role of the transplant coordinator is multifaceted and demanding, requiring advanced clinical skills in addition to patient management responsibilities. Although much is known about the clinical skills of these coordinators, there is virtually no information available about their management abilities. A comparison of the beliefs of transplant coordinators and transplant staff nurses confirms that the coordinators have the role expectations and communication styles to allow them to function effectively in managing patient care as well as in working with physicians and other health professionals. PMID- 9727107 TI - Prevention, early detection, and treatment of breast cancer: a collaborative approach. AB - Breast disorders are not only common and concerning, but in many cases, represent a real risk of death. Dealing knowledgeably with breast-related concerns is a priority for all HCPs and requires a thorough understanding of primary and secondary prevention strategies; first, to reduce the incidence of breast cancer, and second, to reduce the morbidity and mortality associated with the disease. Coordinated and supportive care for women diagnosed with breast cancer is essential to promote optimal health and well-being. The knowledgeable primary HCP, working collaboratively with a breast specialist, can help women achieve these goals. PMID- 9727108 TI - Differential diagnosis of a breast mass. AB - The differential diagnosis of breast masses by the primary care provider should be based on thorough history and physical assessment, age-appropriate diagnostic tools, and careful follow-up. Confident determination of the benign nature of most breast lumps, combined with consistent screening measures, will promote optimum breast health. Additionally, early diagnosis of breast cancer is clearly enhanced by early detection of symptoms and contributes to successful treatment. PMID- 9727109 TI - The significance of nipple discharge: diagnosis and treatment regimes. AB - Breast cancer is one of the most dreaded diseases affecting women and is associated with a high degree of morbidity and mortality. Any breast complaint has the potential for creating a great deal of anxiety for patients and providers alike. An in-depth understanding of the pathophysiology of common breast complaints, particularly those with low probability of association with cancer such as nipple discharge, can serve to allay anxiety and prevent the financial and emotional burden of unnecessary diagnostic evaluations. It will then improve the quality of care for each patient experiencing the symptom. Although the greatest incidence of nipple discharge is not secondary to malignant processes, the fact that some are requires that all persons with nipple discharge receive the benefit of a thorough assessment. When planning a course of action to assess and treat nipple discharge, a thoughtful stepwise, planned approach is necessary. The evaluation of nipple discharge can be undertaken with minimal difficulty by performing a thorough history, a careful physical examination, and following a logical thought process in linking the type of discharge with the suitable adjunct diagnostic testing. Appropriate management evolves from this process. It is important to view the patient in total, considering such issues as family history, other risk factors, how disruptive the level of anxiety about the symptom. Primary care providers, working with their parents, are well positioned to design appropriate diagnostic and treatment regimes to assess and treat nipple discharge. A thoughtful, prudent approach to this symptom can save both health care dollars and lives. PMID- 9727110 TI - Issues in breast imaging. AB - Until there is a cure for breast cancer, clinicians must focus their attention on current modalities of early detection that can accurately diagnose the early stage of the disease. Mammography continues to be the most reliable and accurate tool for breast imaging although controversy continues to obscure the issue of appropriate mammographic screening for women 40-49 years of age. Agencies such as the American Cancer Society and the American College of Radiology recommend year screening for women older than 40 years of age. Patient risk factors should be examined thoroughly in making individual decisions regarding breast screening. Additionally, the historic perspective of mammography and new technologies emerging in improved breast imaging are presented. The radiologist's interpretation of mammography and the role of the primary care provider are also reviewed. PMID- 9727111 TI - Psychosocial aspects of breast cancer. AB - This article provides an overview of the psychosocial aspects of breast cancer. Common psychologic reactions during detection, diagnosis, and treatment include anxiety, denial, anger, and depression. Suggestions for primary care provider interventions such as providing education and support, are discussed. Many treatments for the disease seriously challenge a woman's self-image and sexuality. Family issues are equally important including relationships with children and spouses or partners. Social support can be a powerful force which facilitates successful coping with these challenges. The advent of genetic testing poses a whole set of psychologic and social issues which primary care providers will need to be aware of in the future. Primary care providers have an important role in determining the psychological outcome of breast cancer. By being aware of the common psychologic and social problems, they can be prepared to contribute to a successful outcome. PMID- 9727112 TI - An overview of surgical management of stage I and stage II breast cancer for the primary care provider. AB - Breast cancer is the most common cancer affecting women. With early detection and treatment, most patients survive cancer and lead full, active lives. Two principal treatments for stage I and stage II breast cancer are total mastectomy with axillary node dissection (modified radical mastectomy) and lumpectomy with axillary node dissection (breast conservation). Preoperative evaluation is aimed at the patient's ability to withstand the stress of anesthesia and surgery. Lumpectomy is a low-risk procedure; however, more serious complications may result from axillary node dissection, and wound infection occurs in 4-18% of mastectomy patients. Patient and family education is aimed at the operative experience as well as home care to promote physiological and psychologic adjustment postoperatively. Following mastectomy, the breast cancer patient may be fitted with a prosthesis or consider breast reconstruction with a saline implant, expander, or flap procedure. The primary care provider has the opportunity to establish a therapeutic alliance with the patient by providing coping strategies, support, and education throughout the process. PMID- 9727113 TI - Pediatric and adolescent breast health. AB - The incidence of serious breast pathology in children and adolescents is low. Despite this, children, adolescents, and their parents come to the primary care provider with many breast-related concerns. Tanner staging is a good predictor of ensuing physical sexual development and can be used as a tool to reassure the pubertal girl that she is normal. Variations such as polythelia, premature thelarche, gynecomastia, nipple irritation from exercise, and nipple piercing and tattooing can be managed with education and reassurance. Whereas breast cancer is extremely rare in adolescents, evaluation with ultrasonography may be indicated. The primary care provider plays a central role in educating young girls and adolescents about healthy breast practices such as avoidance of alcohol and cigarette smoking, exercise, low-fat diet, breast self-examination, and avoidance of piercing and tattooing. PMID- 9727114 TI - The medical treatment of breast cancer. PMID- 9727115 TI - Mastitis. PMID- 9727116 TI - Mastalgia. PMID- 9727117 TI - Breast self-exam. PMID- 9727118 TI - Needle localization surgical breast biopsy. PMID- 9727119 TI - A case study of a pregnant woman diagnosed with breast cancer during her third trimester. PMID- 9727120 TI - Nursing shortage: are we there yet, or is it still coming. PMID- 9727121 TI - Total parenteral nutrition in surgical patients. AB - Malnutrition is common in the hospitalized surgical patient. There is a strong association between improved nutritional status and a favorable postoperative recovery, while malnutrition has an adverse effect on surgical outcome (Driscoll & Blackburn, 1990; Souba, 1997). Advances in providing parenteral nutrition have had a positive impact on nutrition care of these patients and will be reviewed in this article. PMID- 9727122 TI - Evaluating physical and behavioral changes in older adults. AB - In older adults, subtle and sometime not so subtle physical or behavior changes can act as early warning signs of changing status. Nonspecific signs and symptoms occurring in older adults such as decline in previous functional capacity, urinary incontinence, anorexia, confusion, or unexplained falls may be signs of infection, medication interaction, dehydration, constipation, or sleep deprivation. Nurses, by critically assessing the situation early, may identify a developing problem. Prompt and early diagnosis of the underlying problem may save costly extended hospitalization or even prevent life-threatening complications. PMID- 9727123 TI - Doing more with less: using silent in-services for staff development. AB - A unit's greatest asset is nurses who are up-to-date in their practice. Time and money constraints demand innovative and creative educational methods. Silent inservices teach and empower while encompassing multiple learning styles in a cost-efficient manner. PMID- 9727124 TI - Relationship between nurse caring and patient satisfaction. AB - A correlational study examined patients' (n = 335) reports of nurse caring and satisfaction with nursing care, using the Caring Behaviors Inventory and Patient Satisfaction Instrument. A strong, positive correlation (r = 0.78, p < .001, R2 = 61.46%) was found. The outcomes of this study have important implications for adult health nurses. PMID- 9727125 TI - A new form of heparin is changing treatment of thrombotic disorders. PMID- 9727126 TI - Barmentoring: mentoring and critical nursing behaviors among novice nurses in clinical nursing practice. AB - The findings of this pilot study provide some support that there is a significant positive relationship between the level of achievement rating of critical case (basic psychomotor skills) and the level of barmentoring (encouragement) of novice nurses in clinical nursing practice. The leadership gained from the developmental refinement of the mentor/protege relationship is a key to acting and thinking as a professional in nursing practice. Although the mentor/protege relationship may contribute to personal and professional goal attainment, mentoring should not be regarded as a panacea, but as one modality for enhancing professional development (Messner, 1991; Yoder, 1990). The limitations of the study include a fairly homogenous sample, self-reported data, and lack of knowledge of the sample's preceptors. The sample size notably affects the generalizability of the findings as well as the statistical findings in the study. PMID- 9727127 TI - Career opportunities for doctoral-prepared nurses. AB - As we face the 21st century, transitions fare occurring in all of society's major institutions. Nowhere is this transition more evident than in education and the health care industry. There is an explosion of information and the use of educational technology is now opening doors never entered before. In health care, greater influence by third-party players and consumers can be seen as major factors in a new and more highly competitive health care system. With the advent of managed health care, clearly more money must be spent in nursing research since in mos instances, nursing is a big ticket item in any health care agency's budget. Determining best practice models and testing them, participating in outcomes research that looks in multidisciplinary practice, and developing new knowledge to serve the needs of our rapidly changing and diverse population requires more and better prepared nurse researchers. The 21st century will also bring unprecedented numbers of nursing faculty who are part of the baby boomer generation face-to face with retirement. The retirement of these doctoral prepared faculty will create a huge vacuum that will demand to be filled. Clearly, given the demographics in our country, as these doctoral-prepared nurses reach retirement age, they and their peers will begin to experience the chronic illnesses that are characteristic of our older citizens. More, not less, professional nurses will be needed to meet the demand for health promotion, disease prevention, and care of the acute and chronically ill. More, not less, nursing faculty will be needed to produce the require number of professional nurses and to replace their retiring colleagues. A doctoral degree in nursing will be the key to many opportunities and will bring greater rewards than those who hold the same degree today. For the next generation of graduates, the doctor degree in nursing will truly be a "hot ticket item". PMID- 9727128 TI - Physical restraints: meeting the standards/improving the outcomes. PMID- 9727129 TI - Valuing the competency of the medical-surgical nurse. PMID- 9727130 TI - Quality: the staff nurse's role. PMID- 9727131 TI - Central venous access--catheters, technology, and physiology. AB - Medical-surgical nurses increasingly encounter central venous access devices. Effective patient care involves both the routine management of these devices as well as identifying potential complications and appropriate treatments. The key physiologic principles research, and clinical practices associated with vascular access technologies are presented. PMID- 9727132 TI - Domestic violence: legal, practice, and educational issues. AB - Domestic violence is a recognized and growing public health concern in the United States. Health care professionals have a duty to improve the identification of victims of domestic violence, intervene effectively, and advocate for better education to break the cycle of abuse. PMID- 9727133 TI - Postoperative nursing care contributions to symptom distress and functional status after ambulatory surgery. AB - The relationship of postoperative patient-perceived nurse caring behaviors to symptom distress and functional status in 100 adult ambulatory surgical patients was examined. These behaviors explained 9.3% to 18.2% of the variance in functional status on the 1st, 4th, and 7th day postsurgery, and 10% of the variance in symptom distress on the 7th postoperative day after controlling for ASA physical status classification, preoperative symptom distress, and preoperative functional status. PMID- 9727134 TI - Radiation and chemotherapy as combined treatment for advanced head and neck cancer. AB - Combining radiation and chemotherapy can be a successful treatment for advanced head and neck cancer. This aggressive approach can prevent surgical complications but carries significant acute and long-term effects. Overall management is multidisciplinary and depends on skilled nursing intervention. PMID- 9727135 TI - Review of nursing home regulations. AB - Nursing homes are complex, specialized institutions for a growing segment of the population. Health care professionals should be aware of government regulations that currently influence nursing home practice. The effects of these regulations as well as consumer implications necessary for optimal care of nursing home residents will be addressed. PMID- 9727136 TI - Intellectual capital. PMID- 9727138 TI - Quality taken for granted. PMID- 9727139 TI - Achieving magnet status. AB - Creating and maintaining a professional practice environment that ensures quality patient outcomes continues to be a challenge in a cost-constrained health care environment. The ability to be recognized for excellence by nursing colleagues is made possible by the Magnet Nursing Services Recognition Program. This article will describe the application preparation, site visit, and evaluation experience of Robert Wood Johnson University Hospital. PMID- 9727137 TI - Quality amid turbulent times. PMID- 9727140 TI - CAPNA: a new development to increase quality in primary care. Columbia Advanced Practice Nurse Associates. AB - An innovative primary care practice managed and staffed by Columbia University School of Nursing faculty opened at Columbia-Presbyterian Eastside in midtown Manhattan on September 29, 1997. The faculty's first commercial venture, Columbia Advanced Practice Nurse Associates (CAPNA), responded to the need to further validate cost, quality, and competence benefits of advanced practice nurses. The aim is to show these benefits exist for patients who are not in the underserved populations historically served by advanced practice nurses. CAPNA is the first faculty group practice in nursing contracted with managed care companies as primary care providers under the same reimbursement arrangements as physicians. PMID- 9727141 TI - St. Joseph's Hospital and Medical Center. AB - To weather the chaos and turbulence in health care and maintain high quality patient services, hospital organizations must have a strong mission and value infrastructure and practice a teamwork philosophy. St. Joseph's Hospital and Medical Center has translated their vision into action. This article describes the foundation and structure that supported creative change in the roles of health care providers, the delivery system, education, and performance improvement. Successful methods are outlined for providing health care to ambulatory through tertiary level patients and from newborns through geriatrics while maintaining the mission to serve the poor. PMID- 9727142 TI - Expanding the role of the pediatric nurse from inpatient to community health. AB - Innovative programs that expand the role of nurses from inpatient to community health nursing benefit the hospital, the nursing staff, and the community. This hospital implemented a grant-funded prevention-oriented program for high-risk children that demonstrated positive outcomes to children and families, Utilization of pediatric nursing staff was maximized through collaboration of inpatient and home care departments. Nurses experienced increased job satisfaction and professional growth. PMID- 9727143 TI - Improving clinical effectiveness through an evidence-based approach: meeting the challenge for nursing in the United Kingdom. AB - Improving clinical effectiveness is a major challenge facing nurses working in the United Kingdom and requires a coordinated approach in order to ensure that the information about which interventions work is made available to those in a position to use it. This means that policy makers, administrators, and nurses need to base decision making on the best available evidence. In this article we explore the background to the drive for evidence-based practice and discuss how a group of nurse researchers have begun working with nurse administrators and practitioners in a large acute hospital to help change the rhetoric of evidence based practice in nursing into reality. PMID- 9727144 TI - Care management the right balance of care and management? AB - In 1990, nursing leadership at St. Peter's Medical Center, realizing a need to be creative in order to meet the cost and quality demands of the ever-changing health care system, developed a case management system model called Care Management. The qualifications for the role and the orientation of the Care Managers is describes changes in the program. Innovative programs integrated into the model include patient pathways, workers' compensation care management, and ambulatory care management. Outcomes described include length stay and cost reductions. Future direction of the program in the continuum of care is reviewed. PMID- 9727145 TI - Quality measures essential to the transformation of the Veterans Health Administration: implications for nurses as co-creators of change. AB - Health care systems are changing at an unprecedented rate, but few are making the changes in a system affecting nearly 200,000 staff in over 1,100 different sites of service delivery originating from 171 medical centers nationwide, as is the Veterans Health Administration. The issues of change, quality of care, morale and opportunities involved in being a nurse today in a system undergoing this magnitude of change is presented within the framework of the quality of care initiatives that have been launched by VA. The new organization design of VA, emphasizing local decision-making, a description of the multiple quality programs recently introduced and integrative strategies that have been used by the Nursing Strategic Healthcare Group, the VA corporate level policy and nursing programs information center for the country, to support the change process are discussed. PMID- 9727146 TI - Information technology: soul of a new organization. AB - If the total quality organization is the structure of the 1990s, then the structure of the new millennium is the world class organization--and information technology (IT) is the foundation upon which it will be built. This article explores how clinical information systems and IT are evolving to support several unique characteristics of the world class organization, including (1) how patient centered care and the advent of the interdisciplinary care team has led to the development of a new breed of clinical information systems; (2) how accountability has empowered nursing and led, in turn, to a need for systems and languages to measure and document nursing care and its outcomes; and (3) how care across the continuum and rapid, constant change demands open systems that are scalable and interoperable. PMID- 9727147 TI - Nasal receptors and reflexes. PMID- 9727148 TI - Treatment of sinusitis in the next millennium. AB - Sinusitis is an increasingly more important disease due to its increasing prevalence, costs, and recognition. Most acute sinusitis episodes follow colds or acute allergic rhinitis. Chronic sinusitis is most commonly due to allergic and nonallergic rhinitis or anatomical defects in the nose. Several common immunologic abnormalities usually present as sinusitis and may be recognized first by the allergist-immunologist. Treatment involves a carefully selected antibiotic prescribed for an adequate period of time, nasal hygiene using nasal saline washes, topical nasal corticosteroids, and decongestants. Medical management of sinusitis tends to be effective, even in patients with long standing sinus disease. PMID- 9727149 TI - Co-existence of asthma and allergic rhinitis: a 23-year follow-up study of college students. AB - The purpose of this study is to examine the co-existence of asthma and allergic rhinitis among former college students who were diagnosed with these diseases either before or after their freshman year. A total of 738 former Brown University students (69% males and 31% females) who were evaluated and underwent skin testing during their freshman year completed a 23-year follow-up questionnaire inquiring of their history of allergies and asthma. The mean age of the participants at the time of the follow-up study was 40 years. In this group, the cumulative incidence of asthma was 11.3% (84/738), hay fever was 41.5% (306/738), and nonseasonal allergic rhinitis was 14.0% (103/738). The cumulative incidence of allergic rhinitis (hay fever) and/or nonseasonal allergic rhinitis (was 45.8% (338/738). Among the 84 individuals with a cumulative incidence of asthma, 63 (75.0%) had a history of hay fever, 27 (32.1%) had a history of nonseasonal allergic rhinitis, and 72 (85.7%) had a history of allergic rhinitis. Among the 306 participants with a cumulative incidence of hay fever, 63 (20.6%) had a history of asthma. Twenty-seven (26.2%) of the 103 individuals with a history of nonseasonal allergic rhinitis had a cumulative incidence of asthma. Among the 338 individuals with a cumulative incidence of allergic rhinitis 72 (21.3%) had a history of asthma. Among the participants with a history of both asthma and hay fever, 44.8% developed hay fever first, 34.5% developed asthma first, and 20.7% developed both diseases at the same time. Among the individuals with a history of asthma and nonseasonal allergic rhinitis, 38.5% developed nonseasonal allergic rhinitis first, 30.8% developed asthma first, and 30.8% developed both diseases at the same time. This study further demonstrates the frequent co-existence of asthma and allergic rhinitis. Among asthmatics, allergic rhinitis occurred in 85.7%. Only 14.3% of asthmatics did not have allergic rhinitis. Among individuals with allergic rhinitis, asthma occurred in 21.3%. Also, allergic rhinitis often precedes or occurs at the same time as asthma. PMID- 9727150 TI - The emotional needs of allergic and asthmatic patients and their families. AB - This article describes the emotional needs of allergic and asthmatic patients and their families that must addressed if optimal management and well-being is to be achieved. Health care professionals need to recognize the psychosocial issues of patients and parents, which include the effect of stressors, illness perceptions, fears, and concerns and that the management of asthma and allergies requires a team effort, involving a wide variety of health care professionals, e.g., physicians, nurses, social workers, psychologists, teachers, as well as involvement of voluntary health organizations and local support groups. Asthma self-management educational programs address these emotional needs and provide the foundation for gaining control. PMID- 9727152 TI - Allergy and nursing. PMID- 9727153 TI - Rhinitis, sinusitis, and polyposis. PMID- 9727151 TI - Stressors and concerns in teen asthma. AB - Adolescence is a time of increased stress because of intellectual, physical, and sexual maturity culminating in a desire for autonomy. Chronic asthma is perceived as a burden handicapping autonomy. This is apparent in the impairment of athletic and social activity. Furthermore, there is the requirement for inhaled medications that are perceived as hampering crucial peer identification. Nonadherence is epidemic, best resolved by empathetic rapport with the adolescent, family, and peer group while maintaining the status of a culturally and ethnically sensitive professional respected by the adolescent. The adept provider negotiates treatment plans in consultation with the adolescent with mutual respect. Treatment needs to be simple; once or twice per day, with a clear action plan acknowledging when to call the provider. PMID- 9727154 TI - Food-induced allergy in childhood. PMID- 9727155 TI - Asthma: a practical overview. PMID- 9727156 TI - Asthma among the famous. Helen Hayes (1900-1993), American actress and author. PMID- 9727157 TI - Asthma among the famous. Harold D. West (1904-1974), American medical scientist and educator. PMID- 9727158 TI - Asthma among the famous. Robert Donat (1905-1958), British actor. PMID- 9727159 TI - Asthma among the famous. Elizabeth Bishop (1911-1979), American poet. PMID- 9727160 TI - [Prospects for control of the occurence of antibiotic resistance]. PMID- 9727161 TI - [Gradient extraction of ammonium specific ionophore antibiotics from mycelium]. AB - Streptomyces brunneofungs 118 and S.griseolus 224 were isolated from natural objects and shown to synthesize ammonium specific products belonging to macrotetrolide compounds. Gradient extraction was applied to the mycelium and it was demonstrated that the compounds were rather labile both in the native cells and on synthetic carriers and could be hydrolyzed by aqueous solutions of acetone and ethanol to various linear oligomers of narctinic acids. Acetone mainly stabilized the monomer and dimer fragments whereas in the ethanol extracts a complete set of the oligomers (from the monomer to the tetramer) was detectable. Graident extract of suspension of the microbial intact cells is useful in the study of some properties and the primary identification of biologically active hydrophobic products even at the early stages of their isolation. PMID- 9727162 TI - [Pyrimidine derivatives increase antibiotic therapy efficacy after ionizing irradiation]. AB - Pyrimidine derivatives increased the antibiotic therapy efficacy in albino rats irradiated with RUM-7 apparatus for close-focus roentgenotherapy. 2-Methyl-4 amino-6-oxypyrimidine was twice as efficient as oxymethyluracil and 6 times as efficient as methyluracil in the stimulation of the skin reparative regeneration. When the total irradiation was performed with LUCH-1 apparatus in a dose of 6 Gy the pyrimidine derivatives also increased the antibiotic therapy efficacy. After the prophylactic use of the pyrimidine derivatives for 7 days prior to the total irradiation their therapeutic effect increased, the level of the exudative component lowered, the tissue epithelization increased, the terms of the wound healing decreased and the animal lifespan increased. PMID- 9727163 TI - [Multicenter study of Staphylococcus susceptibility to antibiotics in Moscow and St. Petersburg]. AB - Antibiotic susceptibility of 898 Staphylococcus strains isolated in 9 medical centres of Moscow and St. Petersburg was tested. The frequency of methicillin resistant staphylococci (MRS) ranged from 0 to 40 per cent for Staphylococcus aureus and from 0 to 65.9 per cent for coagulase negative staphylococci (CNS). The highest frequencies of MRS were in hematologic, oncologic and traumatologic units as well as in intensive care units for newborns. The frequencies of MRS in St. Petersburg were much lower than those in Moscow. Among the methicillin susceptible staphylococci the frequency of the isolates that produced beta lactamases amounted to 81 per cent for S. aureus and to 60 per cent for CNS. Practically 100 per cent of the isolates was susceptible to ampicillin/sulbactam and cephalosporins of the 1st, 2nd and 3rd generations. 86-99 per cent of the isolates was susceptible to erythromycin, clindamycin, tetracycline, gentamicin and trimethoprim/sulfamethoxasole. All the MRS isolates were susceptible to vancomycin. 95, 84 and 70 per cent of the S.aureus strains were susceptible to fusidin, rifampicin and ciprofloxacin, respectively. The susceptibility of the CNS isolates was somewhat lower. PMID- 9727164 TI - [Susceptibility of beta-hemolytic streptococci to antibiotics and alternative drugs]. AB - Antibiotic susceptibility of the circulating beta-hemolytic streptococci of serogroups A, B, C and G isolated from healthy and sick children and adults within 1987-1996 (more than 900 cultures) was studied. It was shown that the MICs of betalactam antibiotics did not change (within the susceptibility levels) with respect to the serogroup A, C and G streptococci. The number of the strains resistant to erythromycin and lincomycin increased the same as the frequency of the strains simultaneously resistant to chloramphenicol, tetracycline and gentamicin. Tomicid and solkarmon, drugs alternative to antibiotics, were found to be active against the above streptococci groups and efficient in the treatment and prophylaxis of streptococcoses. A suggestion that betalactams are not safe in the treatment of the toxic shock syndrome due to the group A streptococci is discussed. PMID- 9727165 TI - [Program of empirical antibacterial therapy of community-acquired pneumonia]. AB - Clinical and bacteriological efficacies of some antibacterial agents were estimated with their differential use in the management of various groups of patients with community-acquired pneumonia. Group 1 included young and middle aged patients with mild pneumonia. Group 2 included young and middle-aged patients with moderate pneumonia. Group 3 included elderly patients with pneumonia and/or patients with concomitant diseases or certain factors complicating the main process. The patients of group 1 were treated with roxithromycin and spiramycin and showed a rapid clinical effect in 100 and 86 per cent of the cases and a rapid bacteriological effect in 84 and 75 per cent of the cases respectively. The patients of group 2 were treated with parenteral cefuroxime with positive clinical and bacteriological effects in 68 and 78 per cent of the cases respectively. The patients of group 3 were treated with ceftibuten and pefloxacin which provided a clinical effect in 91 and 70 per cent of the cases and a bacteriological effect in 72 and 100 per cent of the cases respectively. The results of the treatment with an account of the differences in the pathogen spectra made it possible to recommend as the 1st order agents for the empirical therapy of community-acquired pneumonia (1) macrolide antibiotics for young and middle-aged patients with mild pneumonia without concomitant diseases, (2) 2nd generation cephalosporins for patients with moderate pneumonia without severe concomitant diseases and (3) 3rd generation cephalosporins or fluoroquinolones for elderly patients with pneumonia and the patients with complicating factors. PMID- 9727166 TI - [Pharmacokinetic interaction of fluoroquinolones and other drugs]. PMID- 9727167 TI - Boxing, youth and children. AB - Boxing remains a popular spectator sport, success in which is achieved by the promotion of boxing in the childhood years. Despite the failure to ban boxing generally, there remains the realistic expectation that organised boxing by children below the age of consent (appropriately 16 years) is still achievable. There are two reasons for banning children under the age of 16 years from boxing. The first is that children have little awareness of risk, specifically the risk of chronic encephalopathy, which develops only after a lag period measured in decades or more. The second is that there is no place in contemporary society for a youth sport which has, as its primary goal, the infliction of acute brain damage on an opponent. This paper analyses the medical and ethico-social issues implicit in this subject. Boxing, in historical perspective, has been an altruistic amateur sport for boys and male youths, and has a proud tradition. But in the context of social evolution, and in current perspective to prevent exploitation of underprivileged youths, the time has come for an absolute ban on underage boxing. Such is achievable even if adult boxing remains an Olympic and television sport for some time to come. PMID- 9727168 TI - Prevention of serious bacterial infection in children with nephrotic syndrome. AB - Although nephrotic syndrome is well known as a predisposing factor to bacterial infection in children, especially peritonitis due to Streptococcus pneumoniae, data on the incidence of infection and the effectiveness of preventative measures are limited. With particular reference to pneumococcal disease, this review summarises the available data on the pattern and incidence of invasive bacterial infection in children with nephrotic syndrome, and the level of evidence for the use of penicillin chemoprophylaxis and pneumococcal immunisation. Although data on the effectiveness of pneumococcal immunization in children with nephrotic syndrome are limited, the safety profile of this vaccine makes the risk-benefit ratio favourable to use of the current polysaccharide vaccine in those over 2 years of age. Conjugate pneumococcal vaccines are likely to be more effective, particularly in children under 2 years of age and should be available by the year 2000. Although penicillin prophylaxis against pneumococcal infection is not of proven benefit for nephrotic syndrome, it is beneficial in sickle cell disease without appreciable risk. Subgroups of patients with nephrotic syndrome most likely to benefit from twice daily phenoxymethyl penicillin prophylaxis include children under 2 years of age, with unresponsive or frequently relapsing disease, or who have had a previous episode of pneumococcal infection. PMID- 9727169 TI - Soy protein formula. The Australian College of Paediatrics. PMID- 9727170 TI - Child rearing and cultural beliefs and practices amongst Thai mothers in Victoria, Australia: implications for the sudden infant death syndrome. AB - OBJECTIVE: To examine the perceptions of sudden infant death syndrome (SIDS) and to describe the role of cultural beliefs and practices on child rearing amongst Thai mothers in Victoria, Australia. METHODOLOGY: In-depth interviews and participant observation conducted with 30 Thai mothers during 1995-96. RESULTS: SIDS was not known amongst these Thai mothers prior to migration to Australia. However, they were aware of SIDS when they gave birth here and all of them expressed fear about their baby's death. Due to this fear, most mothers tended to follow Thai beliefs and practices strictly to prevent death. These included breast-feeding, not leaving the infant alone at nighttime, placing the infant on the side or back to sleep, and bedsharing. It is considered that there are numerous evil spirits who may harm the infant, but some are benevolent and protect the newborn, such as ancestral spirits and the guardian angel of a child. Several Thai rituals are carried out to protect the newborn from ill health and death, including inviting the soul of the infant to reside in his or her body, and ritual clipping and shaving the hair of the newborn within the first month of life. CONCLUSIONS: Cultural beliefs, rituals and child-rearing practices help Thai parents to overcome their fear of SIDS. Hypotheses derived from Asian parents' child rearing practices may be useful in further SIDS research. PMID- 9727171 TI - Antibiotic hypersensitivity reactions in cystic fibrosis. AB - OBJECTIVES: To document the frequency and severity of reactions to antibiotics in children and adolescents with cystic fibrosis (CF), to determine which drugs and routes of administration are most likely to produce reactions and to assess how these reactions limit the choice of antibiotics. METHODOLOGY: Medical records were reviewed to ascertain the number and routes of courses of antibiotics and all suspected drug reactions. Patients and their parents were interviewed about drug reactions. RESULTS: Fifty-three records were suitable for analysis. Eighteen of 53 subjects had experienced a reaction (34%). The intravenous route was most allergenic, with 33% of treated patients experiencing a reaction and 9.5% of courses provoking a reaction. Piperacillin was the most allergenic antibiotic. CONCLUSION: Drug hypersensitivity reactions are common in CF. Piperacillin is particularly allergenic. Whilst rarely life-threatening, the reactions are unpleasant and can limit our choices for antibiotic treatment of their bronchopneumonia. PMID- 9727172 TI - Postural drainage in cystic fibrosis: is there a link with gastro-oesophageal reflux? AB - OBJECTIVES: To determine the clinical effects of a change from postural drainage (PD) to positive expiratory pressure chest physiotherapy (PEP) in children with cystic fibrosis (CF) and symptoms of gastro-oesophageal reflux (GOR). To measure the effects of PD on GOR in children with CF. METHODS: Study 1: Six adolescents with CF and symptoms of GOR during PD were changed to upright PEP physiotherapy. The effects on lung function, reflux symptom scores and annual hospital days were measured. Study 2: Twenty-four children with CF (mean age 11 years) and symptoms suggestive of GOR underwent 24-h pH monitoring, including periods of chest physiotherapy. RESULTS: Study 1: All six patients reported a reduction in reflux symptoms during PEP therapy (P < 0.001). Lung function parameters improved during the first 6 months of PEP (P < 0.001). This improvement was sustained for a further 18 months. Annual hospital days decreased significantly (P < 0.0005). Study 2: Nine of 24 patients (37.5%) had pathological GOR. Reflux episodes were significantly increased during PD (P < 0.0001), as was fractional reflux time (P < 0.01). CONCLUSIONS: Upright PEP physiotherapy may be more appropriate than PD in selected patients with CF and symptomatic GOR. The role of GOR as a cofactor in the progression of pulmonary disease in CF needs further evaluation. PMID- 9727174 TI - Measles immunity and immunization status in under-5-year-old children in New South Wales: a population-based study. AB - OBJECTIVE: To estimate the proportion of 1-4-year-old New South Wales children immune to measles and compare the documented immunization history with serologically defined immune status. DESIGN: Population based seroprevalence survey piggybacked onto the National Survey of Lead in Children. Immune status was determined by two different enzyme immunoassays on plasma samples from subjects. SETTING: New South Wales, February-March 1995. OUTCOME MEASURES: Documented measles immunization collected by interview survey and serologically defined immunity. RESULTS: Of 689 survey subjects, 430 (62.4%) provided a blood sample. Adequate plasma remained for both assays for 347 children, of whom 279 (80.4%) were immune by both assays. Parents of 330 stated that their children were immunised, of whom 211 (63.9%) were able to produce corroborating records. Of these 211 subjects, 178 (84.4%) were immune compared to 87 (76.3%) of 114 without records (P = 0.07). CONCLUSIONS: We estimate the prevalence of true measles immunity in 1-4-year-old NSW children to be only 80%, a level inadequate to prevent outbreaks of measles in urban populations. Both long term and immediate strategies are required to increase the prevalence of immunity among NSW children; these may include lowering the age of the routine second measles dose and mounting a mass measles immunisation campaign to include preschool aged children. PMID- 9727173 TI - The use of human glutathione S-transferase A1 in the detection of cystic fibrosis liver disease. AB - OBJECTIVE: To determine the value of serum human glutathione S-transferase A1 (hGST A1) in the detection of cystic fibrosis liver disease (CFLD). METHODS: Sixty-three children (aged 0.5-16 years) with cystic fibrosis (CF) were screened prospectively for evidence of hepatobiliary abnormalities between February 1993 and February 1996. Comparison was made between clinical examination, abdominal ultrasonic scan, measurement of conventional liver enzymes (LFTs) and serum hGST A1 concentration in the detection of hepatobiliary abnormalities in children with CF. RESULTS: The 5-95% concentration of serum hGST A1 was 1.7-4.27 micrograms L-1 for the control group. The hGST A1 levels in the CF patients were significantly higher than in the non-CF group. Thirty-eight (60%) children had detectable hepatobiliary abnormalities. Ultrasound scanning detected the highest number of abnormalities (41%), followed by hGST A1 (30%). The presence of clinical liver disease was found in 19% of the children. The estimated sensitivities of detecting CFLD by clinical method, ultrasound scan, serum hGST A1, and LFTs would be 32%, 68%, 50% and 16%, respectively. CONCLUSIONS: Serum hGST A1 measurement increases the sensitivity of detecting hepatic abnormalities when included with clinical and ultrasound evaluation although, in some cases with advanced liver disease, serum hGST A1 may be normal. Conventional liver enzyme tests add little information in the detection of CF liver disease. PMID- 9727175 TI - A population-based survey of immunization coverage in children aged 2 years and younger in New South Wales. AB - OBJECTIVE: To provide a population-based baseline of immunization rates in children aged 2 years and younger in New South Wales (NSW) in 1992, permitting more accurate evaluation of the efficacy of current programmes. METHODS: A cross sectional population-based survey of 622 households from areas resident to over 73% of all children aged 4 years and younger in NSW. RESULTS: Of the 322 households with children aged 3-24 months, 212 (66%; confidence interval (CI): 57 75%) were up-to-date with the recommended immunization schedule, 68 (21%; CI: 15 27%) had not commenced any immunization, and 42 (13%; CI: 9-17%) were partially immunised. Ability to read English (odds ratio (OR): 5.43; CI: 2.37-12.44) and receipt of hepatitis B immunization (OR; 2.54; CI: 1.27-5.07%) were highly associated with up-to-date immunization; whilst a history of any illnesses, frequent doctor visits in the past 12 months (OR: 0.47; CI: 0.27-0.85%) and older age (16-24 months) (OR: 0.26; CI: 0.12-0.50%) were less likely to be associated with up-to-date immunization. CONCLUSIONS: In 1992 NSW had low levels of up-to date immunization. Significantly, one-fifth of NSW families with children aged 3 24 months did not have a record of any immunizations. This could not be explained by delay in commencing immunization. Poor competency in reading English was strongly associated with failure to immunise, suggesting that there had been inadequate targeting of immunization campaigns in non-English-speaking communities. PMID- 9727176 TI - Immunogenicity and reactogenicity associated with an 18-month booster dose of a new diphtheria-tetanus-whole cell pertussis vaccine. AB - OBJECTIVE: To establish safety and immunogenicity of a reformulated whole cell pertussis based diphtheria-tetanus-pertussis vaccine (DTPw) at the 18-month booster stage following a 2, 4, and 6-month primary immunization course. METHOD: Open trial in suburban Melbourne in 100 healthy children initially recruited through maternal and child health centres. Thirty-five subjects were bled prior to vaccination, and 4-6 weeks after vaccination. A 7-day diary card was used to record subject temperatures and other systemic and local clinical signs. RESULTS: The increase in antibody geometric mean titres (GMT) after boosting was 19.5-fold (95%ci 14.2, 27.2) for tetanus and 26.5-fold (95%ci 16.6, 42.4) for diphtheria. Pertussis antibody GMTs also all showed substantial increases following the booster, with mean fold changes in titre ranging from 7.3 (Agg2) to 31.3 (Fha). Seventeen percent of subjects (95%ci 10%, 26%) experienced axillary temperatures > or = 38 degrees C during the 24-h period following vaccination. Low rates of significant (> 25 mm) injection site redness (13%) and swelling (8%) were recorded at 24 h postvaccination. CONCLUSION: This vaccine was well tolerated by children at 18 months of age, and showed substantial boosting of antibody to all components. PMID- 9727177 TI - Immunization has no effect on arousal from sleep in the newborn infant. AB - OBJECTIVE: To investigate arousal from sleep in infants before and after triple antigen immunization (DTP). METHODOLOGY: Arousal thresholds were determined in both active sleep (AS) and quiet sleep (QS) the day before and the day following the first DTP immunisation in 14 infants. Arousal was induced using a pulsatile puff of air delivered alternately to the nostrils of the infant. RESULTS: Arousal threshold was significantly higher in QS compared to AS in both studies (P < 0.001). Mean arousal threshold in the pre immunization study was not different from that in the post immunization study in either sleep state. Core temperature and heart rate measured at the time of stimulus delivery were both significantly elevated in the post immunisation study (P < 0.001). Sleep duration and number of awakenings from sleep were unaffected by immunization. CONCLUSION: The first triple antigen immunisation has no effect on arousal from sleep in either AS or QS in this group of infants. PMID- 9727178 TI - Hospitalization for pneumonia in children in Auckland, New Zealand. AB - OBJECTIVE: To describe the epidemiology of hospitalizations for pneumonia in children in Auckland, New Zealand. METHODS: A consecutive sample of children hospitalised with pneumonia at the Starship Childrens Hospital from 1 July 1993 to 30 June 1996. Subjects were Pacific Island, Maori, and European/other children aged 0-14 years resident in north, west and central Auckland who were hospitalized with pneumonia. Comparisons were made of the number of hospitalisations by year, ethnicity, age and season; and of the hospitalisation rates by year, ethnicity and age. RESULTS: There were 681 children who were hospitalized with pneumonia during 1993-94, 731 during 1994-95 and 630 during 1995-96. The average annual hospitalization rate was 5.0 per 1000 children aged 0 14 years (95% CI 4.8-5.2). The average annual hospitalisation rate for Pacific Island children was 14.0 per 1000 (95% CI 13.0-14.9), for Maori children 6.7 per 1000 (95% CI 6.0-7.4) and for European/other children was 2.7 per 1000 (95% CI 2.6-2.9). Fifty-three per cent of the hospitalised children were less than 2 years of age. A larger percentage of Pacific Island (61%) and Maori (60%) children were aged less than 2 years compared to European/other (42%) children (P < 0.001). There was marked seasonal variability in the number of hospitalizations, with peaks in hospitalizations corresponding to peaks in positive respiratory viral isolates. CONCLUSIONS: Pneumonia was a consistent cause of hospitalisation for a large number of Auckland children during this 3 year period. Hospitalisation rates and age distribution varied with ethnicity. Hospitalization rates were highest for Pacific Island. intermediate for Maori and lowest for European/other children. Based on these hospitalisation data, pneumonia is a significant cause of morbidity for children in Auckland, New Zealand. PMID- 9727179 TI - Paediatric nephrotic syndrome in Auckland, New Zealand. AB - OBJECTIVE: To review children with nephrotic syndrome in Auckland, New Zealand. METHODS: All children admitted to Auckland Children's Hospital between January 1984 and December 1994 with nephrotic syndrome and a renal biopsy had their charts retrospectively reviewed and the appropriate information summarised. RESULTS: Fifty-seven children biopsied with nephrotic syndrome were available for review. The mean age at diagnosis was 5.4 years, standard deviation (SD) 3.9 years, with mean follow-up of 5.7 years. The histologies of the renal biopsies were: minimal change nephrotic syndrome (MCNS) in 37%, focal segmental glomerulosclerosis in 19%, membranoproliferative glomerulonephritis (MPGN) in 23% and other causes in 21%. Maori children were most likely to have MPGN. Steroid resistance was present in 66% of children. End stage renal failure developed in 26% of patients and chronic renal failure in 4% of patients. CONCLUSION: In this series the proportion of children with MCNS is low and significant numbers of children with nephrotic syndrome progressed to chronic renal failure and end stage renal failure. PMID- 9727180 TI - Outcome of closed head injury in Malaysian children: neurocognitive and behavioural sequelae. AB - OBJECTIVES: To compare the neurobehavioural outcome of children aged 6-12 years with severe closed head injury [sCHI] (coma > 24 h), mild-to-moderate head injury [mCHI] (coma < 6 h) and orthopaedic controls. METHODS: Twenty-nine children in each group, matched for age, sex and ethnicity, were assessed using the Glasgow outcome Scale (GOS), Weschler Intelligence Scale for Children (WISC-III), Movement Assessment Battery for Children (Movement ABC), Wide Range Assessment of Learning and Memory (WRAML) and a standardised neurological examination 6 months post-injury. Parental reporting of pre- and post-injury behaviour was documented using the Child Behaviour Checklist (CBCL). RESULTS: Seven (24.1%) children with sCHI and three (10.3%) orthopaedic controls had residual motor deficits. Three (10.3%) children with sCHI and none in the other groups faced problems with independent ambulation. Twenty-seven (93.1%) of those with sCHI and all children in the other groups had GOS scores of good recovery or moderate disability. Twenty-two (81.5%) sCHI, five (18.5%) mCHI and one (3.7%) orthopaedic control reported a deterioration in school performance. MANOVAS identified a significant injury group effect for performance skills (P = 0.007), verbal skills (P = 0.002), memory and learning (P = 0.001) and motor skills (P = 0.001). Repeated measures ANOVA for pre- and post-injury CBCL scores showed significant differences related to somatic complaints (P = 0.004), problems of socialising (P = 0.003), delinquency (P = 0.004), aggressiveness (P = 0.010), thought (P < 0.001) and attention (P < 0.001). Post-hoc univariate analysis showed the significant differences were between that of the sCHI children and the other two groups. CONCLUSION: Although most sCHI children seemed to have made good physical recovery, there were cognitive, motor, memory and learning difficulties and behavioural problems concomitant with a deterioration in school performance compared with those with lesser or no head injury. This highlights the need for better integrated rehabilitation services to enable a gradual return into mainstream school. PMID- 9727181 TI - Transcatheter occlusion of atrial septal defects: an initial experience with the Amplatzer septal occluder. AB - OBJECTIVE: To discuss the initial experience with the use of a new double disc occluding device, the Amplatzer septal occluder in transcatheter occlusion of secundum atrial septal defects (ASD). METHODS: Transvenous sizing of secundum ASD was performed in five children under general anaesthesia using transoesophageal echocardiographic and fluoroscopic guidance. An Amplatzer septal occluder equal to or minimally larger than the stretched diameter of the ASD was used for transcatheter occlusion of the defect in three suitable patients. Pulmonary balloon valvuloplasty with a 18 mm x 3 cm Mansfield balloon catheter was carried out in one patient with associated pulmonary valvar stenosis in the same procedure setting. RESULTS: Stretched diameters of the defects in the three patients ranged from 14 to 17 mm. Devices of sizes 14, 17 and 17 were deployed through 7F and 8F sheaths, respectively. The upper and lower rims of interatrial septum were more than 8 mm in all patients. All patients had successful occlusion with complete obliteration of the atrial left to right shunting. Simultaneous pulmonary balloon valvuloplasty for the valvar stenosis reduced pressure gradient from 53 mmHg to 22 mmHg across the valve prior to septal occlusion in one patient. No intraprocedural or short-term complication was encountered. CONCLUSIONS: The design of the Amplatzer septal occluder permits ease in loading, delivery, deployment and stable seating of the device. This initial experience shows that Amplatzer device occlusion is feasible, relatively safe and effective and appears to be a viable alternative to surgical closure of secundum atrial septal defects in properly selected patients. PMID- 9727182 TI - Aetiology of childhood proptosis. AB - OBJECTIVE: To retrospectively analyse causes of childhood proptosis and their investigations and treatment. METHODOLOGY: The records of children under 15 years of age presenting with proptosis to The Children's Hospital, Camperdown, Sydney, were reviewed for the period 1983-93 inclusive. RESULTS: Fifty-seven cases of proptosis were found. In order of frequency the causes were; orbital cellulitis 22, thyroid eye disease 8, optic nerve +/- optic chiasm glioma 8, orbital rhabdomyosarcoma 7, metastatic neuroblastoma 4, orbital neurofibroma 3, orbital haemangioma 2, metastatic Ewing's sarcoma 2 and orbital dermoid cyst 1. Treatment depended on the cause (antibiotics, antithyroid drugs, chemotherapy, radiotherapy, surgery) and was multidisciplinary. CONCLUSIONS: The most common cause of proptosis in children presenting to The Children's Hospital, Camperdown, Sydney, was infective orbital cellulitis. The most useful initial investigation was an orbital computed tomography scan. Treatment depended on the cause of the proptosis and was multidisciplinary. PMID- 9727184 TI - The relationship of age to pathology in pelviureteric junction obstruction. AB - OBJECTIVE: To ascertain the age distribution of the different pathological mechanisms which lead to the development of pelviureteric junction obstruction. METHODS: A series of 165 kidneys in 158 children who underwent pyeloplasty for pelviureteric junction obstruction were reviewed. Those 132 renal units (127 children) with uncomplicated pathology were selected for further study. The operative records were reviewed for the underlying cause of obstruction and the age at operation. RESULTS: Obstruction due to extrinsic compression by an aberrant lower pole vessel occurred in an older group (median age 67.3 mo) than those with a narrowing or angulation of the pelviureteric junction (median age 3.1 mo). CONCLUSIONS: Pelviureteric junction obstruction, secondary to a lower pole vessel presents at an older age. Doppler ultrasonography to detect a lower pole vessel may be of benefit in the management of equivocal cases of pelviureteric junction obstruction, particularly in prenatally diagnosed hydronephrosis. PMID- 9727183 TI - Antenatal steroids, condition at birth and respiratory morbidity and mortality in very preterm infants. AB - AIMS: To investigate (1) perinatal factors affecting condition at birth in very preterm infants (23-32 weeks) and (2) the relationship between poor condition at birth and neonatal respiratory morbidity and mortality. METHODOLOGY: Convenience sample (n = 479) drawn from a geographic population inception cohort. RESULTS: Antenatal steroid use reduced the risk of pH < or = 7.20 [Adjusted Odds Ratio 0.29 (95% CI 0.17, 0.50)], one minute Apgar < 4 [0.54 (0.33, 0.88)], and 5 min Apgar < 7 [0.44 (0.23, 0.84)]. Gestational age was significantly related to Apgar score but not cord arterial acid-base status. No other perinatal factor was significant. Poor condition at birth was associated with an increase in the incidence and severity of hyaline membrane disease and death. These effects were lessened in those exposed to steroids. CONCLUSION: Antenatal steroid use is associated with improved condition at birth and reduces the deleterious effects of poor condition at birth on early respiratory morbidity and mortality. PMID- 9727185 TI - Non-typhoid Salmonella gastroenteritis. AB - OBJECTIVE: To study the clinical features of non-typhoid Salmonella gastroenteritis and the incidence, risk factors and outcome of invasive complications in urban Malaysian children. To describe the serotypes of Salmonella species isolated and the pattern of antibiotic susceptibility. METHODOLOGY: Retrospective review of a group of 131 children with non-typhoid Salmonella gastroenteritis seen at the University Hospital, Kuala Lumpur, Malaysia from January 1994 to December 1996. RESULTS: Sixty-seven percent were infants below one year of age. Fever and vomiting were seen in nearly half of children. Seven children (5.3%) had invasive complications: 5 bacteraemia and 2 meningitis. Age below 6 months, fever > 38.0 degrees C, and dehydration on admission were significantly associated with invasive complications. The commonest serotypes isolated were S. enteritidis, S. paratyphi B, and S. bovis morbificans. A total of 94-100% of isolates were susceptible to commonly prescribed antibiotics. CONCLUSIONS: Children with Salmonella gastroenteritis below 6 months of age who are febrile and dehydrated should be treated empirically with antibiotics until the result of blood culture is available. PMID- 9727186 TI - The synergistic effects of stimulants and parental psychotherapy in the treatment of attention deficit hyperactivity disorder. AB - A 5-year-old boy presented with attention deficit hyperactivity disorder (ADHD), oppositional defiant disorder and separation anxiety disorder. The clinical assessment revealed longstanding parent-child relationship problems, ongoing family stress, and a chronic level of low grade depression in the mother. The treatment approach consisted of drug treatment of the child and long-term psychotherapy of the mother. At termination symptoms associated with ADHD were markedly reduced and parent-child relationship problems were no longer evident. It is argued that in a subgroup of children family stress and attachment difficulties may be involved in the development of ADHD. These difficulties should be considered separately in the treatment of children with ADHD, especially if still present after the symptomatic treatment. The treatment outcome raises the question whether or not certain symptoms attributed to ADHD may be reversible, and the long-term adverse outcome of the condition preventable. PMID- 9727187 TI - Suppression of elevated alanine aminotransferase activity in liver disease by vigabatrin. AB - A patient with hepatic cirrhosis due to Alpers disease is described who had a significantly raised plasma alanine aminotransferase (ALT) activity of 247 U/L which returned to normal (18 U/L) shortly after commencing treatment with the anticonvulsant drug, vigabatrin. Previous studies have reported smaller reductions in plasma ALT activity due to vigabatrin, generally in patients with normal liver function. This case demonstrates that vigabatrin may also reduce markedly elevated ALT activity to the normal range in patients with documented liver disease. Plasma ALT activity cannot be used as an index of liver cell damage in patients receiving vigabatrin. PMID- 9727188 TI - Prader-Willi syndrome: a new study of the Australian Paediatric Surveillance Unit. PMID- 9727189 TI - School screening: comparing outcomes for Australian and overseas born children. PMID- 9727190 TI - ADD/ADHD and hypermobile joints. PMID- 9727191 TI - When is conventional ventilation not conventional? PMID- 9727192 TI - Epidural haematoma and stroller-associated injury. PMID- 9727193 TI - Amelogenin dosage compensation in carcinoma of colon, lung, liver and kidney, is not a marker of clonality in males. AB - The analysis of patterns of X-chromosome inactivation is becoming increasingly utilized as a marker of clonal composition of tissues from women. To date, however, no analogous system has been found for the study of clonality in tissue from men. In the current study, the methylation patterns for portions of the amelogenin genes are tested, which are encoded on both the X- and Y-chromosome (AMGX and AMGY). The polymerase chain reaction (PCR) was used to amplify portions of AMGX and AMGY from genomic DNA of carcinomas of the colon, lung, liver and kidney, as well as from matched normal somatic tissues. The amplification target included Alu I methylation sensitive restriction endonuclease sites as well as a 189 bp sequence which is present in AMGX but is absent in AMGY. Polymerase chain reaction amplification of AMGX and AMGY was successful using genomic DNA from both tumour and normal control tissue in 24 of the 26 cases. Pretreatment of genomic DNA with Alu I blocked amplification of AMGX in all cases from both normal tissue and tumour. This indicates that AMGX and AMGY undergo a non-random pattern of methylation in both normal tissues and in tumours, precluding their use as a marker of clonality. Methylation of Alu I sites in AMGY suggests that the amelogenin genes undergo dosage compensation, which raises the possibility that the expression of amelogenin is not restricted to the development of the tooth bud but may also play some other role in various tissues of the body. PMID- 9727194 TI - Analysis of the 16S rRNA gene sequence of the coryneform bacterium associated with hyperkeratotic dermatitis of athymic nude mice and development of a PCR based detection assay. AB - By 16S rDNA sequencing the authors have characterized the coryneform bacteria associated with hyperkeratotic dermatitis (HD) of athymic nude mice isolated from six different outbreaks of the disease in Northern Italy. This analysis has allowed the authors to confirm the classification of the bacteria as Corynebacterium bovis and to develop a 16S rDNA-based polymerase chain reaction (PCR) detection assay. The test was performed directly on the DNA extracted from epidermal swabs. The PCR primers were chosen to match the 16S rDNA sequence fragments which differ most from the other Corynebacterium spp. The test was shown to be both sensitive and specific for C. bovis. Detection of as few as three viable bacterial cells was possible with the use of an oligonucleotide probe in a liquid hybridization assay. PMID- 9727195 TI - Mycoplasma fermentans DNA is infrequently detected in urine specimens from renal transplant recipients. AB - Mycoplasma fermentans is a likely causative agent of HIV-associated nephropathy. In a pilot study, M. fermentans DNA was detected with polymerase chain reaction (PCR) in urine samples from renal allograft recipients; nine (39.1%) out of 23 renal allograft recipients (most of whom had chronic allograft rejection) and none of the 20 controls, were infected with M. fermentans. A cross-sectional study was conducted to investigate the prevalence of M. fermentans in urine samples from renal allograft recipients. Midstream urine samples were centrifuged at 13,000 x g, purified with QIAamp and tested with PCR using RW004/RW005 and an internal control to screen for the presence of inhibitors. Of the 264 participants recruited, 263 completed the questionnaire (172 men, 92 women); 53 had chronic renal allograft rejection, 106 had chronic renal dysfunction without rejection, 69 had a normal renal allograft for more than 3 months and 35 had a renal allograft for less than 3 months. All urine samples yielded positive results for the internal control. Mycoplasma fermentans DNA was detected once i prospectively collected urine samples. The only individual infected with M. fermentans was also seropositive for HIV-1. This study demonstrates that M. fermentans can be at most sporadically detected in urine from patients living with a renal allograft but is not implicated in chronic rejection of allograft. PMID- 9727196 TI - Comparison of culture and acid-fast bacilli stain to PCR for detection of Mycobacterium tuberculosis in clinical samples. AB - The major drawback in effective use of polymerase chain reaction (PCR) for detecting Mycobacterium tuberculosis (MTB) in clinical samples is the presence of PCR inhibitors and unique cell components of the organism that complicate DNA extraction and subsequent PCR amplification. A PCR assay with a unique multistep DNA extraction method that minimizes these problems was compared in a prospective study to acid-fast bacilli stain (AFBS) and culture for detecting MTB in clinical samples. A total of 254 clinical specimens in two separate studies were processed for MTB by these techniques. While PCR and culture were 100% sensitive and specific, culture required up to 8 weeks of incubation and additional time to perform biochemical testing to identify the isolated micro-organism. Acid-fast bacilli stain had a specificity of about 87% and did not differentiate among Mycobacterial species. In contrast, the results from PCR were available within 48 h and did not require additional testing to attain a final result. Polymerase chain reaction was highly reliable for detection and confirmation and interpretation of positive AFBS results. The assay was easy to perform with a turn around time of about 2 days. PMID- 9727197 TI - Discrimination of Bordetella parapertussis and Bordetella pertussis organisms from clinical isolates by PCR using biotin-labelled oligonucleotide probes. AB - A recently developed shared-primer polymerase chain reaction (PCR) was investigated, in an ongoing pertussis surveillance study for discrimination of Bordetella parapertussis and Bordetella pertussis organisms, by using specific biotin-labelled oligonucleotide probes. From a total of 132 samples, 83 were positive by the B. parapertussis specific probe, 33 were positive by the B. pertussis specific probe and 16 samples containing Hemophilus influenzae as a negative control were below threshold by both probes. The shared-primer PCR in combination with specific oligonucleotide probes provides a rapid, sensitive and specific molecular diagnostic tool for future surveillance studies. In addition, it may be used to further investigate whether B parapertussis antigens should be added to acellular pertussis vaccines to protect against B. parapertussis infections. PMID- 9727198 TI - PNA molecular beacons for rapid detection of PCR amplicons. AB - The authors have developed a method for rapid detection of polymerase chain reaction (PCR) amplicons based on surface immobilized PNA-DNA hybrid probes ('molecular beacons') that undergo a fluorescent-linked conformational change in the presence of a complementary DNA target. Amplicons can be detected by simply adding a PCR reaction to a microtitre-well containing the previously immobilized probe, and reading the generated fluorescence. No further transfers or washing steps are involved. The authors demonstrate the specificity of the method for the detection of ribosomal DNA from Entamoeba histolytica. PMID- 9727199 TI - Development of a PCR microplate-capture hybridization method for simple, fast and sensitive detection of Salmonella serovars in food. AB - The authors have developed an easy and rapid detection and identification system for Salmonella spp. in food. The gene inv A was selected as the target sequence. Oligonucleotides derived from conserved regions of this gene were able to exclusively prime the amplification of a 389 bp fragment when Salmonella spp. DNA was used as the template. An internal Salmonella spp. specific DNA probe was used for confirmation of the amplified polymerase chain reaction(PCR)product, by Southern blot or microplate-capture hybridization assay. In this fashion the sensitivity of the method was increased 100-fold (4.5 fg total DNA). To validate the method, a total of 75 food samples were tested. The PCR-microplate capture hybridization assay is easy to perform and much faster than traditional detection methods for Salmonella spp. in food. Hybridization in microtitre plates is more readily observed than in Southern blot and is more sensitive than conventional agarose gel electrophoresis. PMID- 9727200 TI - Construction of an internal control for the detection of Chlamydia pneumoniae by PCR. AB - For the detection of Chlamydia pneumoniae by polymerase chain reaction (PCR) in respiratory samples, an internal control was constructed to monitor the efficiency of amplification in each reaction. The internal control was designed in a way that the same primer pair can be used to amplify the internal control and target DNA. Nasopharyngeal aspirates of children suffering from asthma (> 2 years of age; 24 patients) or bronchiolitis (< 2 years of age; 47 patients) were analysed for the presence of C. pneumoniae, using the internal control in each amplification reaction. Two specimens from asthma patients, both children of 8 years old, were positive for C. pneumoniae. The number of cases studied is too small to draw conclusions regarding the incidence of C. pneumoniae in different age categories in children. PMID- 9727201 TI - Absence of MEN2A- or 2B-type RET mutations in primary neuroblastoma tumour tissue. AB - Specific germline mutations in the RET proto-oncogene predispose to the familial cancer syndromes: multiple endocrine neoplasia (MEN) types 2A and 2B, and familial medullary thyroid carcinoma. Expression of the RET receptor tyrosine kinase is tightly restricted to tumours of neural crest origin, such as neuroblastoma, and neuroblastoma has been observed in RET transgenic mice. Neuroblastoma tumour cell lines transfected with the MEN2A RET gene exhibit spontaneous neuritic differentiation, whereas MEN2B-type RET transfectants demonstrate altered cell adhesion and enhanced metastatic potential. In this study, the authors examined genomic DNA from 26 primary neuroblastoma tumours for MEN2A and MEN2B RET mutations, using restriction enzyme digestion of polymerase chain reaction products as an alternative to direct sequencing. Examination of RET exons 10 (codons 611, 618, 620), 11 (codons 632, 633, 634) and 16 (codon 918) in all 26 tumours revealed no RET mutations. Taken together these data suggest that abnormalities of the RET signalling pathway, rather than oncogenic, MEN2 type RET activation by mutation, may play a role in neuroblastoma tumorigenesis. PMID- 9727202 TI - Direct quantitation of HIV by flow cytometry using branched DNA signal amplification. AB - Adaptation of the branched DNA signal amplification technology to flow cytometry has resulted in a quantitative nuclei-acid assay with significant advantages over the microwell-based format. In this assay, microbeads, rather than microwell plates, are derivatized with nucleic-acid capture probes and the derivatized beads are used to capture single nucleic-acid targets, which then capture fluorescent reporter probes via branched DNA. The assay detects DNA or RNA targets, has a current lower sensitivity limit of 500 human immunodeficiency virus (HIV) RNA molecules and responds linearly to target level from 500 to at least 50,000 molecules. Since microbeads can easily interrogate large volumes, viral lysis and genomic RNA capture can proceed in one step from comparatively large volumes, and sample preparation is greatly simplified compared to the microwell-format bDNA assay. PMID- 9727203 TI - Molecular identification of a Stenotrophomonas species used in the bioassay for erythromycin in aquaculture samples. AB - A bacterial strain isolated from aquaculture pond slurry, which was extremely sensitive to erythromycin, was used to detect erythromycin at levels as low as 0.05 micrograms ml-1 in aquaculture water, sediments and soil samples. Identification of the indicator organism was attempted by 16S rRNA sequencing, biochemical profile, fatty-acid analysis and polymerase chain reaction (PCR). GenBank comparison showed that the 16S rRNA sequence of the strain was similar to those of more than 20 copies of Xanthomonas and Stenotrophomonas. The position of the strain in a phylogenetic tree based on the 16S rRNA gene sequence comparison is in a cluster of Stenotrophomonas. The fatty-acid analysis also showed that the strain is similar to Stenotrophomonas maltophilia. However, the biochemical profile of the strain is most similar to Xanthomonas campestris, except that it can utilize maltose, which is similar to S. maltophilia. Polymerase chain reaction results showed that the strain is different from X. campestris, S. maltophilia and other Xanthomonas species tested. Based on these results, the authors named this strain as Stenotrophomonas sp. strain NCTR. PMID- 9727204 TI - Cellular Retinol Binding Protein 1 (RBP1): a frequent polymorphism, refined map position and exclusion as the Blepharophimosis Ptosis Epicanthus inversus Syndrome gene. PMID- 9727205 TI - [The present status of research and development of original drugs]. AB - The objective of the article is to make readers familiar with contemporary methods of research and development of original drugs and elucidate economic aspects of pharmaceutical research. The contemporary strategy of research and development in the Research Institute for Pharmacy and Biochemistry is discussed in relation to general trends. Pharmaceutical research and development are characterized by high costs which vary in the majority of pharmaceutical companies engaged in the development of new drugs between 12-16% of the total turnover. The author discusses problems associated with the return rate of these investments in conjunction with patent protection and the time taken to develop original drugs. Expedient use of the large financial funds calls for rational management of research and development. The author therefore pays attention to an analysis of rational methods of management of research and development which are applied in the selection of a research programme and of research projects, to modern methods of research and to the organization of research and development. The author deals subsequently in more detail with contemporary methods of research focused on the detection of leading structures, where molecular modelling and technology of combinatorial chemistry are used. By their combination with optimization procedures using statistical data analysis rational design of molecules is made possible. PMID- 9727206 TI - [Trans-fatty acids in the diet and their possible health risks]. AB - Trans fatty acids are unsaturated fatty acids which have at least one double bond in the trans configuration. Their dietary source are some foods of animal origin, but in particular hardened fats and products which contain them. Increased attention is paid to these substances in particular because they are suspected to act as risk factors for ischaemic heart disease. The authors discuss in the submitted review the chemistry, dietary sources, estimated intake and effects of trans fatty acids in the organism. PMID- 9727207 TI - [The effect of insulin in primary hyperaldosteronism]. AB - BACKGROUND: Deteriorated insulin resistance was described in patients with essential hypertension. The objective of the present study was to test at the receptor and postreceptor level the presence of insulin resistance in hypertension with primary hyperaldosteronism. METHODS AND RESULTS: The diagnosis of primary hyperaldosteronism was assessed by means of biochemical and imaging methods in 123 hypertensive patients with a normal glucose tolerance (mean age 49.8 +/- 12.9 years, range 20-65 years, BMI 28.3 +/- 4.8 kg/m2). The blood pressure was monitored for 24 hours by a Spacelab tonometer (systolic BP 168 +/- 21 mm Hg, diastolic BP 103 +/- 9 mm Hg, plasma aldosterone in a recumbent position 426 +/- 472 pg/ml (normal values under 150 pg/ml), concentration of serum potassium 3.6 +/- 0.6 mmol/l. The control group was formed by seven volunteers matched for age and BMI. The patients had a normal basal blood sugar level in the morning (5.0 +/- 0.6 mmol/l), the basal insulinaemia was 19.5 +/- 10.2 mU/l. The insulin resistance was examined using the method of an euglycaemic hyperinsulinaemic clamp on Biostator at an insulin infusion rate of 1 mU/kg/min. Concurrently insulin receptors on red blood cells were assessed. The plasma potassium concentration was maintained by means of insulin receptors on erythrocytes. The potassium plasma concentration was maintained by means of a linear dosage device with potassium at constant physiological levels (after previous supplementation). In patients with primary hyperaldosteronism the authors observed, as compared with healthy controls, a lower glucose consumption during the clamping (glucose disposal rate 18.7 +/- 4.8 vs 29.3 +/- 3.7 mumol/kg/min, p < 0.01), a rise of the metabolic glucose clearance (3.8 +/- 1.5 vs. 7.0 +/- 1.1 ml/kg/min, p < 0.01 and an index of tissue sensitivity for insulin) 23.7 +/- 9.8 vs. 37.5 +/- 11.6 mumol/kg/min per mU/l x 100, p < 0.02). The characteristics of insulin receptors in patients with primary hyperaldosteronism did not differ from normal values. No correlation was found between the plasma concentration of aldosterone and the index of tissue sensitivity for insulin (r = 0.011, NS). CONCLUSIONS: It may be stated that primary hyperaldosteronism is associated with insulin resistance at the postreceptor level. Its pathogenesis has not been elucidated so far and will be the object of future research. PMID- 9727208 TI - [Soluble TNF and IL-2 receptors in patients with breast carcinoma]. AB - BACKGROUND: Cytokines were shown both to enhance tumour growth and formation of metastases and to inhibit proliferation of tumour cells. TNF alpha may mediate apoptosis and necrosis of cancer cells, the exact role of IL-2 remains to be elucidated. Plasma levels of TNF alpha and TNF and IL-2 soluble receptors (sTNF R, sIL-2R) should thus be in some relation to the biological characteristics of the breast cancer. METHODS AND RESULTS: Plasma levels of TNF alpha, sTNF-R I and II and sIL-2R were measured in 31 women with different stages of breast cancer both before the institution of the therapy and after 3 months of the treatment. Plasma levels of both types of sTNF-Rs were higher in patients with breast cancer than in controls (sTNF-R I-2166.6 +/- 568.9 VS. 1121.3 +/- 260.6 pg/ml, p < 0.001, sTNF-R II-3792.8 +/- 958.9 vs. 1996.2 +/- 404.3 pg/ml, p < 0.001) with no significant difference between clinical stages. Plasma levels of both sTNF-R (0.871, p < 0.001) and sIL-2R tightly correlated one with each other. Plasma levels of TNF alpha decreased after treatment (from 3.92 +/- 1.86 to 3.40 +/- 1.15 pg/ml, p < 0.001), but plasma levels of sTNF-Rs and sIL-2R were not influenced by the treatment. CONCLUSIONS: Plasma levels of soluble TNF receptors may thus serve as a non-specific marker of the untreated breast cancer. Their relation to other biologic characteristics of this tumour is not clear. It remains also to be clarified if the long-term treatment leads to the normalization of sTNF-Rs plasma levels. PMID- 9727209 TI - [Treatment of severe hypertension and hypertensive crises]. AB - The author discusses treatment of severe hypertension and hypertensive crisis in the emergency as well as urgent form. It is in the first place a question of pharmacological treatment but in severe hypertension also non-pharmacological treatment is used to supplement effective control of high blood pressure. The author draws attention to the clinical symptomatology and possible complications of these life threatening conditions. He deals also with the most important groups of antihypertensive drugs and drugs for first aid, the effectiveness of different preparations, their mode of administration and anticipated duration of effect. PMID- 9727210 TI - ACT now! Assertive Community Treatment. PMID- 9727211 TI - Harbinger. I: The development and evaluation of the first PACT replication. AB - While Assertive Community Treatment (originally known as the PACT program) is now recognized around the world as an effective model for rehabilitation of persons with severe mental illness, this was not the case 20 years ago. Harbinger of Grand Rapids, in Kent County, Michigan, was the first replication of the PACT model which sought fidelity and included an experimental design for assessing effectiveness. The design and results are presented from an initial 30-month and a follow-up 66-month evaluation of Harbinger. The 30-month evaluation showed significant differences favoring Harbinger vs, the control group on independent living, employment, and client functioning. At 66-months, there were fewer experimental-control group differences. The differences in results are analyzed in terms of design and data collection problems, changes in the treatment environment for the control group, and the longitudinal course of mental illness. The discussion focuses on next steps in ACT research, utilizing program theory to better establish the mechanisms for successful intervention models. PMID- 9727212 TI - Harbinger. II: Deployment and evolution of assertive community treatment in Michigan. AB - Assertive Community Treatment (ACT) is now recognized as the model proven to be most successful in working with clients with long-term, severe mental illness. The first documented research replication study of ACT was Harbinger of Grand Rapids, in Kent County, Michigan. The Harbinger program influenced significant programmatic changes throughout the public mental health system in Michigan. This paper describes this evolution in community mental health locally and why these changes came about. The state-level strategy to implement replications of Harbinger is described, as well as funding and monitoring mechanisms that have now resulted in over 100 successful ACT programs in Michigan. For mental health administrators, the implications discussed include the future of ACT promotion and implementation, within the reality of a managed care framework. PMID- 9727213 TI - Bridging the gap between inpatient and outpatient providers using organizational elements of assertive community treatment. AB - Even among comprehensive local public mental health systems, there remain large gaps in continuity of care following discharge from inpatient settings. The authors describe a modification of the assertive community treatment (ACT) program model that links inpatients to ongoing community-based care, and provide preliminary evidence of its effectiveness as a component in a rationally organized comprehensive system of care. Given the recent trend toward managed Medicaid arrangements, there will be increased pressure to reduce clients' length of stay in ACT programs. State mental health authorities are cautioned to resist allowing managed care contractors to radically change the conditions under which ACT programs operate until there is greater evidence of the effectiveness of alternative approaches. PMID- 9727214 TI - Implementing assertive community treatment programs in rural settings. AB - The authors present a controlled evaluation of a rural adaptation of the assertive community treatment (ACT) model for clients with serious and persistent mental illness (SPMI). Four community mental health settings adopted an ACT model, while a fifth site blended ACT principles with those of the Rhinelander model, another approach to case management for persons with SPMI. A broad array of client and system outcomes were evaluated at 6, 12, and 24 months into the intervention. Twelve-month findings alerted us to potential problems in implementing the treatment model in study year 1; the implementation was qualitatively evaluated and weaknesses were addressed at the beginning of the second treatment year. Small, positive findings at 24 months suggested that the mid-study course correction may have had an impact. We present these findings along with descriptive data on the challenges of implementing complex services models. We give particular attention to describing implementation barriers to mental health services provision that are uniquely rural. PMID- 9727215 TI - The association between program characteristics and service delivery in assertive community treatment. AB - The authors describe the relationship between service intensity and staffing, organizational, client, and site characteristics in 19 programs based on the Thresholds Bridge adaptation of the assertive community treatment (ACT) model. Pearson correlations were examined between 14 program characteristics and intensity of ACT services. Several staffing and organizational attributes were related to service intensity: larger team size, shared caseloads, greater supervisor involvement in direct client services, and assignment of primary responsibility for the client to the team. The potential facilitating relationship between several aspects of team operation and intensive services is discussed as are implications for local implementation of ACT. PMID- 9727216 TI - Fact: integrating family psychoeducation and assertive community treatment. AB - Family Psychoeducation and Assertive Community Treatment are both state-of-the art service systems with rich empirical foundations, demonstrating unusual effectiveness in randomized clinical trials. Recent research suggests a possible additive effect on selected outcomes when the two approaches are integrated. This paper reviews the role of family support and intervention in the care of persons with serious mental illnesses, presents the research literature on psychoeducation, and highlights benefits of merging the multiple-family version of this approach into the work of assertive community treatment programs. PMID- 9727217 TI - Consumers as staff in assertive community treatment programs. AB - The last decade has witnessed the increasing importance of consumers as providers of mental health services. Assertive Community Treatment (ACT) teams and ACT variants, with their emphasis on rehabilitation and support in the client's natural environment, have hosted consumer-professional collaborations. The authors discuss one such program in which an ACT program for homeless mentally ill adults employed consumer advocates (CAs). Consumer advocates were found to have a service profile similar to other staff. Further, there is suggestive evidence that the employment of CAs created a more positive attitude toward persons with mental illness. Issues of role definition, boundaries, support with supervision and the importance of CAs' experiences with mental illness are discussed. PMID- 9727218 TI - Financing assertive community treatment. AB - Assertive community treatment (ACT) is a widely-used intervention for the most severely impaired persons with mental illness. Because it differs from traditional treatment approaches in its philosophy, its organization, and in the clients it serves, financing strategies that are appropriate for standard services may not be optimal for ACT. In developing new payment systems, policymakers must choose between targeted strategies that attempt to influence the treatment process directly and those that establish broad goals for effectiveness, access, and efficiency while allowing providers more latitude in the treatment process. These choices profoundly influence how and to whom ACT is available. PMID- 9727219 TI - Multisystemic treatment of violent and chronic juvenile offenders: bridging the gap between research and practice. AB - The lack of communication between researchers and practitioners has hindered the development of effective interventions for children and adolescents. Recently, however, significant headway in bridging this researcher-practitioner gap has been made due to the emergence of multisystemic therapy (MST) as a treatment approach that combines the rigor of science and the "real world" aspects of clinical practice in treating violent and chronic juvenile offenders and their families in community-based settings. MST addresses the multiple known determinants of delinquency and delivers services in the family's natural environment, with considerable emphasis on treatment fidelity. This article describes MST and provides a case example of how MST treatment principles are applied. PMID- 9727220 TI - Consumer turnover in service utilization patterns: implications for capitated payment. AB - Risk-adjusted capitation rates based on a group's expected cost to a provider have been proposed to create incentives for including people with severe mental disabilities in managed healthcare systems. The most appropriate methods for classifying people according to levels of risk, however, have yet to be determined. This research employed cluster analysis statistical techniques to identify patterns of services delivered to a sample of 4,346 consumers in Ohio. Findings about the extent of client movement in and out of service clusters over time suggest caution in the implementation of risk-adjusted capitation plans, particularly those based primarily on past service utilization. PMID- 9727221 TI - Practice protocols, parameters, pathways, and guidelines: a review. AB - The evolution of clinical practice protocols is described within the context of its origins in utilization review and utilization management. Physician concerns and barriers to implementation, as well as the role of the Agency for Health Care Policy and Research (AHCPR) are discussed. An example of competing guidelines from the AHCPR and the American Psychiatric Association is described in detail. The available literature on cost savings related to utilization management and clinical guidelines is reviewed and summarized. With over 1,800 medical practice guidelines catalogued, practice protocols have become ubiquitous and continued research into their cost and impact on quality are needed. PMID- 9727222 TI - The efficacy of involuntary outpatient treatment in Massachusetts. AB - One means to address some of the unintended consequences of the shift of treatment for individuals with serious mental illness from hospitals to communities has been involuntary outpatient treatment (IOT). Using Massachusetts data, 19 patients with court orders for IOT were matched to all and to best fits on demographic and clinical variables, and then to individuals with the closest fit on utilization before the IOT date. Outcomes indicated the IOT group had significantly fewer admissions and hospital days after the court order. The full impact of IOT requires more study, particularly directed toward IOT's effects on insight and quality of life. PMID- 9727223 TI - Rural models for integrating primary care and mental health services. AB - This paper presents findings from a study designed to identify and describe models for integrating primary care and mental health services in rural communities. Data were obtained from telephone interviews with staff at rural primary care sites around the country. Findings are based on the responses of 53 primary care organizations in 22 states. The authors identify four integration models--diversification, linkage, referral and enhancement--which appear to exist in combination, rather than as pure types. The proposed analytic framework outlines aspects of integration that are readily amenable to study. PMID- 9727224 TI - Organizational features affecting the use of coercion in the administration of psychiatric care. AB - Historically, the use of coercion in psychiatric hospital admissions, and research on such use, have reflected social circumstances that impact on psychiatric care. Currently, the social emphasis on cost-saving in the U.S. and corresponding shifts in the organization, financing, and management of psychiatric and mental health care, have begun to affect research on the use of coercion in psychiatric admissions. Such research has begun to incorporate hospital organizational dynamics which affect the use of coercion in these admissions. The authors propose that this emphasis should be expanded into a comprehensive research agenda that examines the most pertinent organizational features affecting the use of coercion in psychiatric hospital admissions. PMID- 9727225 TI - Perceived shortages of community support services. PMID- 9727226 TI - County mental health directors' perspectives on forensic mental health developments in New York State. PMID- 9727227 TI - Offenders with mental illness: prevalence and responsibility. PMID- 9727228 TI - The environmental trends facing state mental health agencies. PMID- 9727229 TI - Managed behavioral health and academia: is there a "fit" between services and training. Introduction. PMID- 9727231 TI - Training the trainees: the new imperatives. PMID- 9727230 TI - Training the existing workforce. PMID- 9727232 TI - Training the trainers. PMID- 9727233 TI - A survey of what clinicians should know. PMID- 9727234 TI - Observations. PMID- 9727235 TI - Capital budgeting techniques. PMID- 9727236 TI - Detection of renal arteries with fast spin-echo magnetic resonance imaging. AB - With the increasing use of non-invasive imaging with MR and volumetric CT to evaluate renal arteries, the ability to accurately detect the number and state of native renal arteries becomes critical if conventional angiography is to be supplanted in these settings. The present study evaluated the utility of a fast spin-echo (FSE) T2-weighted sequence to detect the number and course of renal arteries and their ostia compared to conventional angiography. Ten patients underwent conventional catheter angiography either for renal artery stenosis evaluation or as potential renal donors. Each patient then had an MR study of the renal arteries and kidneys with FSE MR (TR = 4000 ms, TE = 102 ms, eight-echo train length, 5-mm-thick interleaved 128 phase encodes, superior and inferior saturation pulses, number of excitations (NEX) = 4, on a 1.5-T superconducting magnet (GE, Milwaukee, WI, USA). Images were reviewed by two 'blinded' radiologists and renal arteries were counted and their ostia were evaluated. Results were compared with angiography and inter- and intra-observer statistics were calculated. All 10 patients underwent MR successfully, nine for renal artery stenosis (RAS) evaluation and one was a renal donor. A total of 24 renal arteries were imaged in 19 kidneys. Fast spin-echo MR is 95% accurate (95% CI: 88-100%) in detection of renal arteries, with no statistical difference between FSE MR and catheter angiography (McNemar P = 0.0). Inter- and intra-observer statistics demonstrate good-to-excellent agreement in renal artery detection (kappa: 0.63 0.90). In one case of RAS evaluation an incidental adrenal mass was detected as the aetiology of the patient's hypertension. Fast spin-echo MR can be a useful adjunct as part of the imaging for renal arteries with MRI. PMID- 9727237 TI - Computed tomography of abdominal neurogenic tumours. AB - Abdominal and pelvic neurogenic tumours are uncommon neoplasms in adults apart from those tumours found in patients with neurofibromatosis. Malignant degeneration occurs in 2.4-29% of neurofibromatosis. Biopsy of neurofibromas can be complicated by sensorimotor nerve deficit. Distinction of malignancy by imaging may circumvent biopsies of asymptomatic benign neurogenic tumours. Benign neurogenic neoplasm is suspected on CT scan if the tumour is in the region of known nerve ganglia or pathway, and is well demarcated, solid, homogeneous, hypodense relative to muscle, and enhances with contrast material. Malignant neurogenic tumours are often large, irregular, infiltrative, and necrotic with heterogeneous contrast enhancement. Computed tomography is valuable in distinguishing malignant from benign neurogenic neoplasms, predicting resectability, detecting distant metastases, and evaluating response to treatment. PMID- 9727238 TI - Longitudinal tibial stress fractures. AB - The tibia is a fairly common site for stress fractures of both the fatigue and insufficiency types. The majority are transverse or, less frequently, oblique. The present report outlines four cases of vertical stress fractures, an orientation that is very uncommon and leads to bone scan findings which can be misleading. PMID- 9727239 TI - The role of the barium enema in the diagnosis of colorectal neoplasia. AB - The recent interest in guidelines for colorectal cancer diagnosis, management and in screening has important implications for radiologists. The present article reviews the role of the barium enema in colorectal neoplasia diagnosis in symptomatic patients, and in the context of screening programmes. PMID- 9727240 TI - Audit of barium enema examinations in the diagnosis of colorectal carcinoma. AB - Barium enema is a commonly used technique. A number of papers have been published describing its accuracy in the diagnosis of colorectal carcinoma. A retrospective comparison of pathology and radiology reports was made to determine rates of demonstration and reporting of colorectal carcinoma on barium enema examinations for a 12-month period. PMID- 9727241 TI - Implementation of image-guided large-core needle biopsy of the breast on a limited budget. AB - Image-guided large-core needle biopsy (LCNB) of the breast is becoming widely accepted as an accurate, minimally invasive and economical way to obtain a tissue diagnosis of breast lesions. However, much of this work has been done with expensive dedicated equipment, often favouring stereotaxic guidance. Image-guided LCNB was introduced to Middlemore Hospital based around existing inexpensive equipment, and stereotaxic or ultrasound guidance was chosen depending on which showed the lesion best. Multidisciplinary clinical, radiological and pathological assessment of each case was performed. The results of 213 biopsies (112 stereotaxic and 101 ultrasound guidance) are reported here. Malignancy was shown or suspected in 85 biopsies (40%). The absolute sensitivity for malignancy was 97% with complete sensitivity of 100%. The absolute specificity was 100% and the complete specificity 98.5%. Large-core needle biopsy can be successfully implemented in a large institution without investment in expensive equipment while maintaining high ratios of malignant/benign biopsies. Attention to technique and careful multidisciplinary review are important to the success of such a programme. PMID- 9727242 TI - Radiologically placed hepatic artery catheter allows selection of patients with high-volume liver metastases for regional chemotherapy. AB - Regional chemotherapy has achieved high response rates in hepatic metastases from colorectal cancer and has been shown to improve survival significantly. The present paper reports the use of pre-operative regional therapy to establish marker response as a means of selection of patients for surgery. Fourteen patients underwent radiologically placed hepatic artery catheter (HAC) for chemotherapy. In the 11 patients with carcino-embryonic antigen (CEA) fall the patient proceeded to open surgical placement of HAC. The predictive effect of CEA fall following radiological HAC was good. Non-responding patients are clearly spared the discomfort and inconvenience and costs of an unnecessary operation. PMID- 9727243 TI - Set-up variation of patients treated with radiotherapy to the prostate measured with an electronic portal imaging device. AB - The set-up variation of 11 patients treated supine with radical radiotherapy for carcinoma of the prostate was measured with an electronic portal imaging device to determine the adequacy of set-up techniques and current margins, as well as the need for immobilization. During the treatments 172 images of the anterior fields and 159 images of the left-lateral fields were taken and the errors in treatment placement were measured by template matching. The variation in the superior-inferior direction was small, 1.4-1.6 mm (1 SD), while the medio-lateral variation was 2.8 mm (1 SD). The anterior-posterior variation was largest, 4.6 mm (1 SD) with an offset of 3.3 mm anterior. This anterior offset and large anterior posterior variation suggests that set-up techniques were not optimal for this direction. The 1 cm margin used was adequate for set-up variation except in a small number of cases, which was mainly due to the anterior trend. Random (treatment-to-treatment) variations were small (1.1-2.3 mm; 1 SD), indicating that immobilization would result in only modest improvement in reproducibility for these supine patients. PMID- 9727244 TI - Delayed presentation of primary testicular seminoma. AB - A case is reported here of delayed presentation of primary testicular seminoma, 9 years after initial presentation with retroperitoneal disease. The diagnostic difficulty associated with primary extragonadal germ cell tumour is emphasized. PMID- 9727245 TI - Rothmund-Thomson syndrome and tolerance of chemoradiotherapy. AB - Rothmund-Thomson syndrome (RTS) is a rare disorder with a predisposition for cutaneous and non-cutaneous malignancy. It is speculated that ultraviolet (UV) sensitivity and deficient DNA repair may account for this predisposition and influence the tolerance of chemoradiotherapeutic management. A case is reported of the management of an RTS patient with squamous cell carcinoma of the tongue who demonstrated increased radiosensitivity and tissue intolerance to chemotherapy. PMID- 9727246 TI - Clinical radiohypersensitivity screening using radiation-induced chromosomal aberrations. AB - Severe acute toxicity to radiotherapy (radiohypersensitivity) can limit the effective use of radiotherapy and it is not usually possible to identify such individuals before treatment commences. Five patients with acute radiohypersensitivity (RH) were detected over a 3-year period. All five RH subjects demonstrated a significantly higher degree of radiation-induced chromosomal aberrations (ICA) in fresh blood lymphocytes when compared to normal controls. Results indicate the feasibility of using the ICA assay in conjunction with other tests to screen for radiosensitivity. PMID- 9727247 TI - Letters from the radiation oncologist: do referring doctors give a damn? AB - Letters sent to referring and associated doctors by a radiation oncologist after consultation and treatment for 128 consecutive patients were reviewed. Overall, only 60% of information items previously identified as 'essential' were included in these letters. An anonymous survey of the referring doctors and general practitioners (GPs) for these 128 patients was conducted. A total of 93 of 103 respondents considered letter content to be satisfactory or very good, nine considered letter content to be only average and only one respondent considered content to be unsatisfactory. The possible reasons for high levels of satisfaction regarding correspondence by referring doctors and GPs in spite of the relatively low level of information content are explored. PMID- 9727248 TI - Cutaneous metastases from adenocarcinoma of unknown primary. AB - Cutaneous manifestations of malignancy are not uncommon, especially in advanced disease. They may also occur early in malignant disease or they may even signify recurrence particularly if they are paraneoplastic in nature. Clinical diagnosis can be difficult because of the wide spectrum of appearance of these lesions, and, in many cases, because of the lack of an identifiable underlying primary. Presented here is the case of a 65-year-old woman with multiple inflammatory cutaneous metastases, which were sclerodermoid in nature. These appeared 14 months after initial diagnosis of adenocarcinoma of unknown primary (ACUP) and signified the beginning of a rapid deterioration in her condition. The coexistence of limited systemic sclerosis (scleroderma) and ACUP initially raised several interesting diagnostic possibilities. Adenocarcinoma of unknown primary and the sclerodermoid reaction in malignancy are discussed. PMID- 9727249 TI - Two case reports: carcinoma of the cervix and carcinoma of the endometrium treated with radiotherapy after previous irradiation for benign uterine bleeding. AB - In the 1940s, 1950s and 1960s, low doses of radiotherapy were used to treat benign uterine bleeding. The cases of two women who received this form of therapy and later developed gynaecological malignancies and had high-dose pelvic radiotherapy are presented. A 76-year-old woman with an International Federation of Gynecology and Obstetrics (FIGO) stage-IIB squamous cell carcinoma of the cervix received external beam radiotherapy and intra-uterine brachytherapy and a 77-year-old women with a FIGO stage-IB endometrial adenocarcinoma received adjuvant postoperative pelvic radiotherapy. Both women had a significant past history of low-dose-rate intra-uterine irradiation for dysfunctional uterine bleeding. Therefore the theoretical question of carcinogenesis was raised, and also the practical questions of what dose had previously been given and what further dose could be safely given with regard to normal tissue tolerance. PMID- 9727250 TI - Extensive intracranial calcification secondary to hypoxia, presenting with dyspraxic gait. AB - The imaging appearances of a case of extensive intracranial calcification presenting with dyspraxic gait are described. Computed tomography showed bilateral calcification in the anterior, posterior and central watershed areas and in the basal ganglia. It is believed that the changes are secondary to previous hypoxaemia and hypotension, and that subsequent development of the symptoms is due to calcification in the dystrophic tissue. PMID- 9727251 TI - Arterial haemorrhage following instillation of silver nitrate in chyluria: treatment by coil embolization. AB - Chyluria is a urological manifestation of lymphatic system disease. Sclerotherapy of the renal pelvis (RPIS) using 1% silver nitrate, along with diethyl carbamazine, is the treatment most frequently used. Massive haematuria due to intrarenal aneurysm following RPIS has not been reported. A case is described here of arterial haemorrhage following instillation of silver nitrate which was treated by coil embolization. The haematuria was immediately stopped and the renal function returned to normal gradually. PMID- 9727252 TI - Iatrogenic gallstones: a ceftriaxone complication. AB - The authors report a case of biliary colic secondary to gallstones formed during a course of therapy with Ceftriaxone. Three weeks after cessation of therapy the gallstones dissolved. PMID- 9727253 TI - Spontaneous arterial thrombosis in association with pancreatic carcinoma: diagnosis and interventional management. AB - A 58-year-old woman presented with bilateral upper limb ischaemia. Digital subtraction angiography demonstrated upper limb arterial thrombosis, extensive on the left. This was initially treated with catheter-directed thrombolysis which cleared the thrombus on the right, although the left upper limb required supplementary surgical embolectomy. Subsequently, biopsy-proven pancreatic adenocarcinoma was detected. The upper limb arterial thrombosis appeared to be spontaneous in association with the malignancy because there was no obvious embolic source. PMID- 9727254 TI - Axillary lymph node calcification due to metastatic papillary carcinoma. AB - A case is presented in which calcification in an axillary lymph node seen on a screening mammogram was the presenting feature of metastatic papillary carcinoma, presumed to be of thyroid origin. The differential diagnosis of axillary lymph node calcification seen on mammography is reviewed. PMID- 9727255 TI - Brachytelephalangic chondrodysplasia punctata. AB - A case is reported here of a boy with brachytelephalangic chondrodysplasia punctata. This is the first case of this disorder reported in the Australian literature. PMID- 9727257 TI - Neuroradiologic findings in brown snake envenomation: computed tomography demonstration. AB - A case of fatal brown snake (Pseudonaja textilis) envenomation is presented. The CT examinations show rapid development and progression of atypical bilateral intracerebral haematomas (ICH) which produce a fatal outcome despite correction of the underlying coagulopathy. The striking CT appearance suggests coagulopathy and is predictive of increased mortality. PMID- 9727256 TI - Malignant ascites visualized on a radionuclide bone scan. PMID- 9727258 TI - Huge adrenal haemangioma. AB - Adrenal tumours are either functioning or non-functioning. Non-functioning adrenal tumours are generally asymptomatic and usually of enormous proportions at the time of presentation. A case is presented here of a patient with a huge right adrenal haemangioma which was successfully treated surgically. This unusual tumour was 25 cm in diameter, was well encapsulated and weighed 4 kg. The literature pertaining to this interesting case is reviewed. PMID- 9727259 TI - Splenic metastasis in germ cell tumour. AB - Metastasis to the spleen from carcinomas is a rare event. A case is described here of a 28-year-old man with germ cell tumour of the testis in whom splenic metastasis was demonstrated radiologically. The lesion resolved with chemotherapy and the patient has been in complete remission for 1 year. PMID- 9727260 TI - Hyperprolactinaemia: don't forget hypothyroidism! AB - Female patients with mild clinical symptoms signs of hypothyroidism may be overwhelmed by the accompanying manifestations of hyperprolactinaemia. Radiological imaging may show enlargement of the pituitary gland. In these cases a primary diagnosis of hypothyroidism may not be considered. Long-term bromocriptine mesylate treatment or surgery can be avoided if the diagnosis is considered and the thyroid function test performed. PMID- 9727261 TI - Congenital tuberculosis. AB - Congenital tuberculosis is a rare disease. The non-specific nature of presenting signs and symptoms (because of the lack of host response) and the fatal outcome in the absence of early therapy all underscore the importance of early diagnosis and treatment in infants. Recognition requires awareness that tuberculosis at this age has manifestations not found in older children. Here a case of congenital tuberculosis is presented, where changes were confined only to the thorax. Tuberculosis in the mother could be diagnosed only retrospectively. PMID- 9727262 TI - Torsion of the wandering spleen. AB - A case of torsion of the wandering spleen with splenic infarction is reported. The medical imaging studies performed with typical findings are described. Case reports have been reviewed. Although rare, torsion of the spleen remains an important differential diagnosis in patients presenting with acute abdominal pain. Early intervention is necessary to reduce the risk of splenic infarction and other complications, and an increased awareness of the condition together with the use of appropriate medical imaging can lead to the correct diagnosis more readily. PMID- 9727263 TI - Post-traumatic intra-osseous pseudomeningocele of the occipital bone. AB - Clinical, radiographic, and operative findings in two children with intra-osseous pseudomeningocele of the occipital bone due to cranial trauma are presented here, along with a literature review of this uncommon radiologic entity. PMID- 9727264 TI - Mistaken angiographic diagnosis of traumatic aorto-atrial fistula. AB - A case of angiographically occult traumatic aorto-caval fistula masquerading as aorto-right atrial fistula is presented. The importance of angiographic vigilance in the imaging of arterio-venous fistula is emphasized. PMID- 9727265 TI - Fluid-fluid levels in a simple bone cyst on magnetic resonance imaging. AB - An unusual case is presented here of simple bone cyst (SBC) with fluid-fluid levels on MR and cementum-like substance on microscopy in an atypical location in the distal femur. Fluid-fluid levels are commonly described in the literature within aneurysmal bone cysts, giant cell tumour, chondroblastomas and telangiectatic osteosarcomas, but a literature review revealed only one reported case with multiple fluid levels occurring in a simple bone cyst on MRI. A cementum-like matrix is diagnostic of SBC and is seen in approximately 10% of cases. PMID- 9727266 TI - Fracture epiphyseal separation of the distal humerus. AB - Seven patients seen with fracture separation of the distal humerus epiphysis have been analysed for the problems linked with the radiological diagnosis of this injury. Peculiar male predominance, exclusive left-side involvement, consistent postero-medial displacement of the epiphyseal fragment and ability to achieve near anatomic reduction by closed manipulation in fresh cases have been some of the other features observed. The literature has been briefly reviewed for this infrequent and usually misdiagnosed injury. PMID- 9727267 TI - Foetal intracardiac transfusion for the treatment of severe anaemia due to human parvovirus B-19 infection. AB - Intra-uterine parvovirus infection may result in severe foetal anaemia and death. Ultrasound diagnosis of foetal parvovirus is presented, together with ultrasound guided foetal transfusion to treat the anaemia. PMID- 9727268 TI - Radiology of the parapharyngeal space. AB - The parapharyngeal space (PPS) is a central space in the deep neck. Intrinsic lesions within this space are limited. Other spaces in the neck are closely related to the PPS and the direction of displacement of this space often suggests the origin of a lesion. The morphology of a lesion, together with the site of origin, helps in narrowing the diagnostic possibilities. The behaviour of the PPS often helps to determine whether a lesion is an intrinsic abnormality or whether it arises from a neighbouring space. Such information is crucial in the planning of surgical approach and the placement of drains. PMID- 9727269 TI - Case quiz. Superior sagittal sinus and left transverse sinus thrombosis. PMID- 9727270 TI - Cranial cystic epidermoid: report of two cases and review of the literature. PMID- 9727271 TI - Follow-up to management strategies for candidates for protease inhibitors and requiring treatment for Mycobacterium tuberculosis. PMID- 9727272 TI - Statewide surveillance for ehrlichiosis--Connecticut and New York, 1994-1997. PMID- 9727273 TI - Pain management. PMID- 9727274 TI - Interest in alternative medicine by first year medical students at the John A. Burns School of Medicine. PMID- 9727275 TI - Chart audit of inpatient treatment of schizophrenic patients: implications for development of coordinated care paths. AB - This study offers important information regarding the standard of care provided to schizophrenic patients treated at one inpatient facility. The findings were particularly useful in the development of a care path for this specific population. Areas for improvement identified in this research include medical tests, master treatment planning, documentation of care, patient teaching, and discharge planning. Given the limited health care dollars and the lack of a cure for schizophrenia, this research emphasizes the fact that treatment guidelines need to be aggressively tested as to their relevance to practice. PMID- 9727276 TI - Admissions, length of stay, and discharge barriers at the Hawaii State Hospital. AB - Based on data gathered from patients, psychiatrists, and social workers at the Hawaii State Hospital, it was determined that the majority of patients had been in the hospital for more than one year, were committed for forensic reasons, and did not need continued hospitalization. An inter-agency systems approach is needed to address the issue of length of patient stay. PMID- 9727277 TI - Cytological assessment of bronchoalveolar lavage: a valuable diagnostic tool in pulmonary diseases. PMID- 9727278 TI - Diagnosis of pulmonary tuberculosis by polymerase chain reaction for MPB64 gene: an evaluation in a blind study. AB - Polymerase chain reaction (PCR) has been found to be a sensitive and rapid method to confirm a clinical diagnosis of tuberculosis. We evaluated PCR for M. tuberculosis complex specific MPB64 gene for the diagnosis of pulmonary tuberculosis, in a double blind study. One hundred and eighty-two clinical samples (sputum, bronchioalveolar lavage and pleural fluid) from patients with a clinical diagnosis of pulmonary tuberculosis and 72 samples from patients with non-tubercular pulmonary lesions and normal healthy individuals were included. The samples were coded and clinical details were concealed from the laboratory, where conventional diagnostic methods and PCR were carried out independent of each other. On decoding and analysing the data, PCR was positive in 59% of single sputum samples from clinically diagnosed pulmonary tuberculosis, while M. tuberculosis could be grown in 18% of the samples. PCR could identify M. tuberculosis in 81.8% of the culture positive sputum samples. PCR was also positive in 71.4% of bronchioalveolar lavage (BAL) fluid and 60.7% pleural fluid samples from clinically suspected cases, which were mostly culture negative. On comparison with response to treatment, PCR was positive in 79.5% of patients who improved on anti-tuberculosis treatment, with a positive predictive value of 92%. PCR for MPB64 gene provides a useful alternative for the diagnosis of pulmonary tuberculosis from sputum and paucibacillary samples like BAL and pleural fluid in which conventional methods show low sensitivity, especially in areas from which strains show a low copy number of other PCR targets like the IS 6110 insertion sequence. PMID- 9727279 TI - Role of circulating inflammatory cytokines in patients during an acute attack of bronchial asthma. AB - Experimental studies show the unique aspects of cytokines profiles in various inflammatory diseases of the lung lead to different clinical manifestations. To elucidate the potential role of cytokines in the pathogenesis of bronchial asthma, plasma interleukins-1 beta, interleukin-6, interferon-gamma and tumour necrosis factor-alpha were measured in 32 asthmatics during an onset of acute asthma. Nine healthy volunteers were included as controls. Cytokine levels were measured by using commercially available ELISA kits. Our results showed that except for interleukin-6, increased concentrations of cytokines were not detected in the controls. Detectable concentrations of IL-6 and TNF-alpha were more common in patients than in controls. However, Interferon-gamma concentrations were below the threshold of detection in both patient and control groups. In conclusions, our results suggest that IL-6 and TNF-alpha are involved during the onset of an acute attack of asthma once the threshold limit has been passed. Hence, these two cytokines are important markers of the inflammatory components of acute asthma. PMID- 9727280 TI - Pattern of secondary acquired drug resistance to antituberculosis drug in Mumbai, India--1991-1995. AB - A retrospective observational study was conducted to find out whether secondary acquired drug resistance to isoniazid and ethambutol is high and to rifamycin and pyrazinamide is low, as is commonly believed in India. There were 2033 patients, whose sputum samples (6099) were reviewed from a specimen registry of the microbiology laboratory for the years 1991 to 1995. Of these, 521 (25.6%) patients [335 males and 186 females; age ranged from 11 to 75 years] had sputum positive culture and sensitivity for acid-fast bacilli (AFB). The drug resistance patterns in our study were: isoniazid (H) 15%, rifamycin (R) 66.8%, pyrazinamide (Z) 72.2%, ethambutol (E) 8.4%, streptomycin (S) 53.6%, cycloserine (C) 39.2% kanamycin (K) 25.1% and ethionamide (Eth) 65.3%. The resistance to streptomycin showed a significant fall over a year while there was a rise in resistance to cycloserine and kanamycin which is significant. The rate of secondary acquired resistance of isoniazid and ethambutol was low, and the rate of secondary acquired resistance to rifamycin and pyrazinamide was high, which is contarary to the common belief regarding these drugs in India. This implies that isoniazid is still a valuable drug in the treatment of multidrug resistance in India. PMID- 9727281 TI - Pulmonary function abnormality in patients with portal hypertension with or without chronic liver disease. AB - Pulmonary function (FVC, FEV1, PEFR, MMEF) and arterial blood gases (ABG) were analysed in 30 patients of portal hypertension. The aetiology of portal hypertension included cirrhosis of liver (n = 10), non cirrhotic portal fibrosis (NCPF, n = 10) and extrahepatic portal vein obstruction (EHPVO). Ten patients with chronic active hepatitis (CAH) without portal hypertension were also studied. Most pulmonary functions were abnormal (low) in portal hypertension and the most affected parameters, were FEV1, PEFR and MMEF (p < 0.05). The same was also observed in CAH, although in less number of patients. Hypoaxemia (26.7%) and wide alveolar--arterial oxygen gradient were observed most frequently in patients of portal hypertension. These patients also had a more alkaline blood pH. EHPVO patients had better lung function and arterial blood gas values. Patients with NCPF had greater impairment in pulmonary function. PMID- 9727282 TI - Allergic bronchopulmonary aspergillosis. AB - Allergic bronchopulmonary aspergillosis (ABPA) is an immunologically mediated lung disease which occurs predominantly in patients with asthma, and is caused by hypersensitivity to colonized Aspergillus fumigatus. It is a chronic, relapsing disorder which can clinically range from mild asthma to fibrotic lung disease. The immunopathogenesis of the disease is not clearly understood. Early diagnosis and aggressive therapy with oral corticosteroids can prevent the development of fibrotic lung disease and are therefore, the cornerstone of management. PMID- 9727283 TI - An unusual lung mass. PMID- 9727284 TI - Chronic post traumatic thoracic aortic aneurysm mimicking a benign cyst. AB - Aneurysm of thoracic aorta may mimic cystic lesions like bronchogenic cyst or hydatid cyst. We report a case of chronic post traumatic thoracic aortic aneurysm in a young lady, initially diagnosed as benign cystic lesion. MRI scan of chest confirmed the diagnosis and subsequently surgical removal of aneurysm was done. PMID- 9727285 TI - Unusual intrapulmonary foreign body: a pencil. AB - A case of unusual intrapulmonary foreign body in the form of a pencil is described. The penetration occurred following an insignificant fall. The diagnosis was made on CT scan and subsequently surgical removal was undertaken successfully. PMID- 9727286 TI - Tricuspid valve endocarditis following elective abortion. AB - Tricuspid valve endocarditis (TVE) following non-septic abortion without an evidence of pelvic inflammation and clinically audible murmur is rare. We report here two cases of TVE following elective abortion. PMID- 9727287 TI - Hydatid disease in children. AB - Although hydatid disease is an uncommon condition in children, yet it needs to be considered in the differential diagnosis of children presenting with homogenous round opacities on chest radiography and hepatomegaly. Six such cases are reported. PMID- 9727288 TI - Health challenges for the coming millennium: a paradigm for success. PMID- 9727289 TI - The entry of African-American students into US medical schools: an evaluation of recent trends. AB - A need to reassess US medical schools' admission of African-American students exists based on recent challenges to affirmative action. The Association of American Medical Colleges (AMMC) provided US medical school enrollment data and characteristics. Measures of enrollment were constructed for each medical school and aggregated by ownership type and state. After peaking at 1311 students in 1994, African-American medical school matriculation decreased by 8.7% by 1996. This decline was disproportionately generated by public medical schools. However, it was not limited to institutions that are located in states where anti affirmative action policies have been implemented. Several schools were consistently successful (e.g., UCLA, Case Western, and Robert Wood Johnson) or unsuccessful (e.g., Texas Tech and Texas A&M) in enrolling African-American students. Recent gains in the enrollment of African-American students are being reversed, particularly at public institutions. Implications exist, particularly for the health of poor and underserved communities that are more likely to be cared for by such students during their careers as physicians. PMID- 9727290 TI - Injection therapy for management of stenosing tenosynovitis (de Quervain's disease) of the wrist. AB - Stenosing tenosynovitis of the first dorsal compartment of the wrist (de Quervain's disease) is a common cause of radial wrist and hand pain and disability. Nonoperative management of the disease has been thought to provide only temporary relief of pain and swelling, while surgical intervention has been viewed as a more definitive treatment. This retrospective study examines the results of 58 patients who presented with de Quervain's disease from January 1993 through December 1995 and were initially treated with a combination steroid/lidocaine injection. After 1 to 3 years of follow-up, 35 patients had complete relief of symptoms with a single injection, 14 had relief of symptoms with two injections, 2 are still under observation, and 7 patients had to undergo operative management. These results indicate that in most patients with de Quervain's disease, treatment with a steroid/lidocaine injection can provide complete relief of symptoms. PMID- 9727291 TI - Predictors of infant mortality among college-educated black and white women, Davidson County, Tennessee, 1990-1994. AB - Strategies to reduce US infant mortality rates often focus on the black-white disparity in rates. Linked Infant Birth and Death Files for Davidson County, Tennessee, from 1990 through 1994 were used to determine infant outcomes for infants born to college-educated white and black women. Risks for adverse outcomes were identified by comparing infant deaths to live births using logistic regression analyses. The following variables entered the logistic model process: maternal and paternal age; race and education; nativity status; maternal risk factors; interpregnancy interval; parity; infant gender; tobacco or alcohol use; number of prenatal visits; trimester in which prenatal care began; marital status; gestational age; and birthweight. After adjustment for the effects of the other variables, a gestational age < 28 completed weeks of gestation was the most significant independent predictor of infant death. Black race was not identified as a significant predictor of infant mortality. Regardless of race, a decrease in infant mortality rates among college-educated women in this country depends on the prevention of preterm births. Strategies to diagnose early preterm labor must proceed from a comprehensive maternal care program for all women. Open channels of communication between patient and provider will form the cornerstone for preterm prevention-intervention programs. Analysis of state and local infant mortality data may identify regional differences in infant mortality rates and differences in risk factors associated with adverse infant outcomes. PMID- 9727292 TI - Acute renal failure in pregnancy: a review of clinical outcomes at an inner-city hospital from 1986-1996. AB - Acute renal failure is a serious complication of pregnancy. Over the past few decades, the overall incidence of acute renal failure in pregnancy has decreased in Western societies. In less developed countries, the incidence of acute renal failure in pregnancy has remained high. This retrospective study examined the incidence, morbidity, fetomaternal mortality, and renal prognosis among pregnant inner-city patients. Serum creatinine levels of all pregnant patients seen at an inner-city hospital from January 1986 to December 1996 were reviewed. Twenty-one consecutive cases of acute renal failure were identified for an incidence of 2 in 10,000 pregnancies. Maternal mortality was high (15.7%) as was morbidity, with a tendency toward a high rate of intrauterine fetal growth retardation. These results suggest that the outlook for acute renal failure in inner-city patients is dismal in sharp contrast to the prognosis for other patient groups with acute renal failure in pregnancy in Western societies. Preventive strategies should be aimed at this subpopulation with a view to improving early prenatal care as well as enhancing overall access to the health-care system. PMID- 9727293 TI - Highly destructive perianal Crohn's disease. AB - This article reports a case of highly destructive perianal Crohn's disease in a 15-year-old boy who presented with fecal impaction and incontinence. Both upper and lower gastrointestinal tract endoscopy were unrevealing. Treatment with intravenous prednisolone and broad-spectrum antibiotics supplemented by enteral feeding with an elemental diet resulted in prompt recovery. However, healing of his perianal lesions began only after a diverting colostomy. Awareness of this uncommon entity is important because prompt recognition can lead to early institution of appropriate treatment and avoid further morbidity. PMID- 9727294 TI - Attracting theorems for cyclic sets and impermanence. AB - We derive attraction theorems for a cycle connecting nonempty compact (isolated) invariant sets using an average Lyapunov function. An application is given to a concrete system having such a cycle with the object to obtain the impermanence of the system. PMID- 9727295 TI - Stochastic epidemics: the expected duration of the endemic period in higher dimensional models. AB - A method is presented to approximate the long-term stochastic dynamics of an epidemic modelled by state variables denoting the various classes of the population such as in SIR and SEIR model. The modelling includes epidemics in populations at different locations with migration between these populations. A logistic stochastic process for the total infectious population is formulated; it fits the long-term stochastic behaviour of the total infectious population in the full model. A good approximation is obtained if only the dynamics near the equilibria is fit. PMID- 9727296 TI - A state space model for the HIV epidemic in homosexual populations and some applications. AB - In this paper we have developed a state space model for the HIV epidemic in homosexual populations which have been divided into subpopulations according to sexual activity levels. In this model, the stochastic dynamic system model is the stochastic model of the HIV epidemic in terms of the chain multinomial model whereas the observation model is a statistical model based on the observed AIDS incidences. This model is applied to the San Francisco homosexual population for estimating the numbers of susceptible people, infective people and AIDS cases and for estimating the probabilities of HIV transmission from infective people to susceptible people given sexual contacts. The results show that the estimated numbers of AIDS incidence trace closely the observed numbers indicating the usefulness of the model. It is observed that the estimated numbers of latent people show multimodal curves and that HIV infection takes place during the primary stage and very late stage. The results have further shown that there are significant differences between the observed AIDS incidences and the estimates by the embedded deterministic model. These results indicate that using the embedded deterministic model to estimate the HIV-infected people and to predict future AIDS cases can be very misleading in some cases. PMID- 9727297 TI - Optimal harvesting under stochastic fluctuations and critical depensation. AB - We consider the derivation of the optimal harvesting strategy maximizing the expected cumulative yield from present up to extinction, under the assumption that the harvested population fluctuates stochastically and is subjected to an Allee-effect. By relying on both stochastic calculus and the classical theory of linear diffusions, we derive both the optimal harvesting thresholds at which harvesting should be initiated at full capacity and the value of the optimal strategy. In contrast to ordinary models which are absent of critical depensation, we show that the presence of an Allee-effect leads to the introduction of a lower harvesting threshold at which the population should be immediately depleted under the optimal policy. Moreover, we demonstrate that discounting increases the incentives to harvest and, therefore, increases the probability of a soon extinction of the harvested population. PMID- 9727298 TI - A sequential algorithm for the non-linear dual-sorption model of percutaneous drug absorption. AB - A sequential algorithm is developed for the non-linear dual-sorption model developed by Chandrasekaran et al. [1,2] which monitors pharmacokinetic profiles in percutaneous drug absorption. In the experimental study of percutaneous absorption, it is often observed that the lag-time decreases with the increase in the donor concentration when two or more donor concentrations of the same compound are used. The dual-sorption model has sometimes been employed to explain such experimental results. In this paper, it is shown that another feature observed after vehicle removal may also characterize the dual-sorption model. Soon after vehicle removal, the plots of the drug flux versus time become straight lines on a semilogarithmic scale as in the linear model, but the half life is prolonged thereafter when the dual-sorption model prevails. The initial half-life after vehicle removal with a low donor concentration is longer than that with a higher donor concentration. These features, if observed in experiments, may be used as evidence to confirm that the dual-sorption model gives an explanation to the non-linear kinetic behaviour of a permeant. PMID- 9727299 TI - Managed care, medical technology, and health care cost growth: a review of the evidence. PMID- 9727300 TI - Hospital conversions from for-profit to nonprofit status: the other side of the story. PMID- 9727301 TI - Do black elderly Medicare patients receive fewer services? An analysis of procedure use for selected patient conditions. PMID- 9727302 TI - Nursing facility staffing in the states: the 1991 to 1995 period. PMID- 9727303 TI - The rash that kills. PMID- 9727304 TI - Premedical prerequisites revisited. PMID- 9727305 TI - Brown University School of Medicine, Class of 1998. PMID- 9727306 TI - Physicians newly licensed in Rhode Island during 1997. PMID- 9727307 TI - Rapid onset tardive dyskinesia ("fly catcher tongue") in a neuroleptically naive patient induced by risperidone. PMID- 9727308 TI - Adult immunization. PMID- 9727309 TI - Contraceptive methods and utilization in Rhode Island. PMID- 9727310 TI - Palliative care for cancer patients--current issues. PMID- 9727311 TI - [Drainage of the urinary tract as preparation for extracorporeal lithotripsy]. AB - Upper urinary tract drainage in patients with chronic calculous pyelonephritis (CCP) results in not only successful anti-inflammatory and antibacterial treatment but also in more effective and safe ESWL. In 21 CCP patients with upper urinary tract drainage by means of catheter-stent, ESWL was performed using Lithostar-Plus (Siemens). Active inflammation with marked pyuria, bacteriuria and even moderate upper urinary tract dilation were indications for the upper urinary tract drainage with catheter-stent before and during ESWL in CCP patients. Upper urinary tract drainage with catheter-stent contributed to effective treatment of chronic pyelonephritis and allowed to perform ESWL. There were neither attacks of acute pyelonephritis nor upper urinary tract obstruction after catheter-stent removal. The catheter-stent allows to create closed drainage system with active evacuation function as it functions in physiological conditions. ESWL in patients with upper urinary tract drainage using catheter-stent is more effective and has lower risk of complications. PMID- 9727312 TI - [Functional state of the upper urinary tract during and after extracorporeal lithotripsy]. AB - Ureteral motility reaction to extracorporeal lithotripsy was studied in patients with urolithiasis. This reaction depended on the initial function of the urinary tracts. Multichannel impedance ureterography provides objective information on ureteral function. Individual approach is necessary in the treatment of urolithiasis patients. PMID- 9727313 TI - [Ways of prevention of kidney lesions during nephrolithotomy or extracorporeal lithotripsy in nephrolithiasis]. AB - Outcomes of nephrolithiasis treatment depend not only on renal dysfunction resultant from abnormal urine passage and concomitant infection but also on intraoperative mechanical trauma and ischemia. To compare renal damage induced by nephrolithotomy with that of shock-wave lithotripsy and effects of chemotherapy, 54 patients with nephrolithiasis were divided into 2 groups and 2 subgroups. Patients of group 1 underwent nephrolithotomy, those of group 2--extracorporeal shock-wave lithotripsy (ESWL). Subgroups "a" in both groups received no chemotherapy, subgroups "b" were treated with alpha-tocopherol, pentoxifylline, indomethacin. ESWL was found less traumatic. Postoperative drug prophylaxis promoted more pronounced inhibition of lipid peroxidation products and urinary enzymes activity, contributing to lessening of the operative trauma. Anti ischemic protection of the kidneys also improved renal function. PMID- 9727314 TI - [Changes in renal and testicular veins in left-sided varicocele and choice of the surgical method in children and adolescents]. AB - Selective phleborenotesticulography, ++tensiometry of the left iliac, right and left renal veins, vena cava inferior, duplex scanning of the left renal vein were performed in 356, 296 and 57 patients, respectively, of a total of 356 examinees aged 8-17 years with left-side varicocele. Among other tests were measurements of hormones in the blood from the left and right testes (n = 24), pO2, pCO2. Stenosis, aortomesenteric compression (AMC) of the left renal vein, left-side venous renotesticular hypertension (RTH) of the left renal vein were diagnosed in 12, 342 and 158 patients, respectively. Secondary genesis of left varicocele has been proved. The diagnosis of left-side phlebohypertensive nephropathy was made preoperatively. Estradiol content in the blood of the left testis was 1.7 times higher than from the right one. 4 groups of patients were formed: group 1 patients (n = 142) had stenosis, AMC, dilatation of the testicular vein (DTV) and RTH; group 2 patients (n = 18) had AMC, DTV, borderline high pressure; group 3 patients (n = 18) had AMC, RTH, multiple thin testicular veins; group 4 patients (n = 174) had moderate AMC without hypertension. 160 patients of group 1 and 2 have undergone two-direction venous testiculo-iliac anastomosis operation. 193 patients of groups 3 and 4 have undergone Ivanissevich's operation. After Ivanissevich's operation 2 patients with secondary varicocele of the third degree retained varicocele of the first degree. After establishment of anastomoses, neither varicocele recurrences nor anastomosis thrombosis were registered. PMID- 9727315 TI - [Evaluation of the functional state of the urinary tract in children with congenital hydronephrosis]. AB - The study of intrarenal and upper urinary tracts urodynamics in the presence of the ureteropelvic obstruction and of the function of the lower urinary tract shows that congenital hydronephrosis was in 42.8% of cases associated with bladder dysfunction presenting with hyperreflex-type dysfunction. Maladapted hyperreflex bladder was diagnosed in 29.4%, adapted--in 13.5% of cases. The hyporeflex dysfunction occurred in 15.9% of cases. The urinary bladder dysfunction in children with congenital hydronephrosis accounts for 58.7%. These aggravate the same defects of the upper urinary tracts and deteriorate the course of chronic obstructive pyelonephritis. PMID- 9727316 TI - [Effects of plasmapheresis and lasers on the state of urinary microflora in chronic pyelonephritis]. AB - Bacteria isolated from the urine of 142 patients with chronic pyelonephritis (CPN) were examined for pathogenic properties of the strains (bacteriuria, hemolytic, proteolytic properties, urease, adhesive activity, antibiotic resistance, the ability to inactivate bactericidal activity of the serum) to control the effect of the treatment: antibiotics combined with plasmapheresis or antibiotics combined with laser radiation (intravascular, transcutaneous, or both). Combined application of intravascular and transcutaneous laser irradiation in multimodality treatment reduces the number of highly pathogenic strains as well as antibactericidal activity of the urine strains. It also promotes normalization of bacteriuria level. Plasmapheresis is inferior to laser radiation but ranks the second in efficacy of action on urinary microflora. Thus, use of efferent methods, especially transcutaneous plus intravascular laser radiation, plasmapheresis, in combined treatment of pyelonephritis decreases pathogenicity of urine strains and normalizes bacteriuria. PMID- 9727317 TI - [Urine chemiluminescence in preclinical diagnosis of neonatal drug-induced nephropathy]. AB - Animal experiments demonstrate that gentamycin, kanamycin and carbenicillin affect lipid peroxidation in the kidneys. This can be registered by chemiluminescence of renal and urinary homogenates. This phenomenon was also used in functional examination of the kidneys in 161 neonates treated by antibiotics. The earliest renal dysfunctions due to nephrotoxicity were detected by chemoluminescence induced by ions of bivalent iron. The method is simple and rapid, this making it convenient in neonatal practice for detection of nephropathy before the symptoms presentation. PMID- 9727318 TI - [Morphofunctional state of ischemically damaged kidney transplant in preservation and reperfusion (an experimental study)]. AB - In cadaveric renal transplantation with simple cold storage, pretransplant evaluation of allograft viability creates an important problem. To assess the degree of ischemic damage and microcirculation of the renal allograft, experiment comparing parameters, like median perfusate flow and median intrarenal resistance, with histological findings was made. Kidney harvesting was performed from three groups of New Zealand rabbits by keeping warm ischemic time zero, thirty, sixty minutes for each group. Following perfusion with Euro-Collins solution and 24 and 48 hours of hypothermic preservation, perfusion was repeated. Perfusion parameters were measured and histopathological examination of the kidneys was made with light microscopy. In arterioles and glomerular, peritubular capillaries, red blood cell trapping was seen. It was also observed that an increase of warm ischemic time aggravates the erythrocyte aggregation and tubular degeneration. Following reperfusion, resistance measured from renal arteries by Protocol Propaq 106 manometer was named as terminal intrarenal resistance. It is noticeable that this simply measured parameter shows correlation with other parameters and histopathological findings. PMID- 9727320 TI - [Long-term results of proscar (finasteride, MSD) treatment of patients with benign prostatic hyperplasia]. AB - Patients with benign prostatic hyperplasia (BPH) received proscar (finasteride) in daily dose 5 mg for: at least 6 months (n = 206, 20.4%), 12 months (n = 513, 50.9%), 24 months (n = 164, 16.3%), 3-4 years (n = 125, 12.4%). Because the size of the prostatic gland reduced significantly (by 22%) only after one year of proscar treatment, the duration of this treatment should be continued for at least 12 months, in some patients as long as the whole life. The duration of the treatment should be decided for each individual patient. The improvement after the proscar treatment was observed in 95% of the patients treated for benign prostatic hyperplasia. PMID- 9727319 TI - [Changes in the levels of prostate-specific antigen and its molecular forms with alpha 1-antichymotrypsin in patients with benign prostatic hyperplasia]. AB - Analysis of the changes in the levels of total prostatic specific antigen (t-PSA) in patients with benign prostatic hyperplasia (BPH) shows that the patient's age, size of the prostatic gland and chronic bacterial prostatitis influence the levels of t-PSA but have no effect on the levels of PSA-ACT. The relationship between the levels of t-PSA and age in BPH patients is explained by growing mass of benign hyperplasia causing mechanical load on the intact prostatic tissue. The maximal concentration of t-PSA of 8.7 +/- 1.22 ng/ml was observed in BPH patients at the age of 61-70 years. BPH stages, chronic pyelonephritis, chronic non bacterial prostatitis, chronic renal failure are not essential for t-PSA and PSA ACT and can be neglected in interpretation of t-PSA values in BPH patients. PMID- 9727321 TI - [Features of transurethral electric resection of the prostate in patients undergoing thermal treatment]. AB - 23 patients with benign prostatic hyperplasia (BPH) aged 60-82 years underwent transurethral resection (TUR) of the prostate in different periods after thermal treatment which had appeared uneffective or brought complications. In the performance of the endoscopic techniques we found macroscopic changes of the prostatic parts of the urethra and bladder cervix characteristic for certain thermal impact (energy, power, site of exposure). Intraoperative bleeding of prostatic tissue was also different depending primarily on the time which had passed after the thermal treatment. Minimal bleeding occurred at least 3 months after the thermotherapy. Thus, thermal treatment of the prostate can be used in combined treatment of BPH for reducing intra- and postoperative hemorrhage due to subsequent TUR. Among the methods of thermal therapy, transurethral microwave thermotherapy is preferable as minimally invasive and deeply penetrating into the depth of the prostatic gland with maximal effect. TUR of the prostate should be performed not earlier than 3 months after thermotherapy which is indicated only for patients at high risk of intraoperative hemorrhage because of unaffected circulation. Therefore, it is desirable to include transrectal dopplerography of the prostate to urological examination of BPH patients. PMID- 9727322 TI - [Adrenal pseudocyst]. PMID- 9727323 TI - [Extraperitoneal cyst of the right spermatic cord]. AB - A rare case of an extraperitoneal cyst of the right spermatic cord is reported. Dysfunction of the right kidney diagnosed initially appeared to be due to compression of the low third of the right ureter by an extraperitoneal cyst. After removal the cyst the function of the right kidney completely recovered. PMID- 9727324 TI - [A rare and little known variant of hypospadias]. AB - Hypospadia is a congenital defect presenting as different variants of dysplasia of the distal urethra with ectopy of the urethral external orifice and distortion of the penis. A variant of hypospadia not yet described in Russian medical literature was observed in a newborn boy. He had normal penis and scrotum, but on the under-side of the penis there was a site of thin skin protruding as a resonance bag in urination. At the site of the thin skin the back urethral wall was absent with a cavity covered by the thin skin. In urination urine filled the cavity blowing up the skin above it to the form of the resonance bag. This variant of hypospadia was designated as segmental urethral hypoplasia. PMID- 9727326 TI - [Ultrasonography in the diagnosis of obstructive uropathies]. PMID- 9727325 TI - [One-stage bilateral uretero-cystoneostomy and plastic surgery of vesicovaginal fistula]. PMID- 9727327 TI - [Ambulatory monitoring of urodynamics in the diagnosis of urination disorders in women]. PMID- 9727329 TI - Chest pain. What to expect in the ER. PMID- 9727328 TI - Evaluation of certain veterinary drug residues in food. Forty-eighth report of the Joint FAO/WHO Expert Committee on Food Additives. AB - This report presents the conclusions of a Joint FAO/WHO Expert Committee convened to evaluate the safety of residues of certain veterinary drugs in foods and to recommend maximum levels for such residues in food. The first part of the report considers standards for the performance of studies, residues at the injection site, and several initiatives to promote transparency of the process for setting Maximum Residue Limits (MRLs). A summary follows of the Committee's evaluations of toxicological and residue data on a variety of veterinary drugs: two anthelminthic agents (moxidectin and tiabendazole), eight antimicrobial agents (ceftiofur, danofloxacin, dihydrostreptomycin, streptomycin, enrofloxacin, flumequine, gentamicin and spiramycin), one glucocorticosteroid (dexamethasone), and two insecticides (cyfluthrin and fluazuron). Annexed to the report are a summary of the Committee's recommendations on these drugs, including Acceptable Daily Intakes and MRL's and further toxicological studies and other information required. PMID- 9727330 TI - Health tips. Caring for blisters. PMID- 9727331 TI - New guidelines mean more Americans are overweight. PMID- 9727332 TI - Living with chronic illness. Redefining "normal". PMID- 9727333 TI - Rosacea. Are you red in the face? PMID- 9727334 TI - Lycopene. Another good reason to eat tomatoes. PMID- 9727335 TI - When I woke up this morning, part of the white of my left eye was blood red. I didn't injure it in any way, it doesn't hurt and my vision is unchanged, but it looks awful. Should I see my doctor? PMID- 9727336 TI - My 8-year-old granddaughter gets terrible headaches. Do children get migraines? What can we do for her? PMID- 9727337 TI - Mechanical blood-tissue interaction in contracting muscles: a model study. AB - A finite element (FE) model of blood perfused biological tissue has been developed. Blood perfusion is described by fluid flow through a series of 5 intercommunicating vascular compartments that are embedded in the tissue. Each compartment is characterized by a blood flow permeability tensor, blood volume fraction and vessel compliance. Local non-linear relationships between intra extra vascular pressure difference and blood volume fraction, and between blood volume fraction and the permeability tensor, are included in the FE model. To test the implementation of these non-linear relations, FE results of blood perfusion in a piece of tissue that is subject to increased intramuscular pressure, are compared to results that are calculated with a lumped parameter (LP) model of blood perfusion. FE simulation of blood flow through a contracting rat calf muscle is performed. The FE model used in this simulation contains a transversely isotropic, non-linearly elastic description of deforming muscle tissue, in which local contraction stress is prescribed as a function of time. FE results of muscle tension, total arterial inflow and total venous outflow of the muscle during contraction, correspond to experimental results of an isometrically and tetanically contracting rat calf muscle. PMID- 9727338 TI - Steady flow dynamics of prosthetic aortic heart valves: a comparative evaluation with PIV techniques. AB - Particle Image Velocimetry (PIV), capable of providing full-field measurement of velocities and flow stresses, has become an invaluable tool in studying flow behaviour in prosthetic heart valves. This method was used to evaluate the performances of four prosthetic heart valves; a porcine bioprostheses, a caged ball valve, and two single leaflet tilting disc valves with different opening angles. Flow visualization techniques, combined with velocity vector fields and Reynolds stresses mappings in the aortic root obtained from PIV, and pressure measurements were used to give an overall picture of the flow field of the prosthetic heart valves under steady flow conditions. The porcine bioprostheses exhibited the highest pressure loss and Reynolds stresses of all the valves tested. This was mainly due to the reduction in orifice area caused by the valve mounting ring and the valve stents. For the tilting disc valves, a larger opening angle resulted in a smoother flow profile, and thus lower Reynolds stresses and pressure drops. The St. Vincent valve exhibited the lowest pressure drop and Reynolds stresses. PMID- 9727339 TI - Dynamic knee loads during gait predict proximal tibial bone distribution. AB - This study tested the validity of the prediction of dynamic knee loads based on gait measurements. The relationship between the predicted loads at the knee and the distribution of bone between the medial and lateral sides of the tibia was examined. The motion and external forces and moments at the knee were measured during gait and a statically determinate muscle model was used to predict the corresponding forces on the medial and lateral tibial plateaus. In particular, the relationship between the knee adduction moment during gait and the ratio or distribution of medial to lateral tibial bone mineral content was studied. Bone mineral content was measured with dual energy X-ray absorptiometry in four regions, two proximal regions 20 mm in height, one medial and one lateral and two distal regions 20 mm in height, one medial and one lateral. The best single predictor of the medial lateral ratio of proximal bone mineral content (bone distribution) was the adduction moment (R2=0.31, p=0.003). Adding weight (negative coefficient. p=0.0004) and the ratio of the average predicted peak force on the medial plateau to the predicted peak force on the lateral plateau (positive coefficient, p=0.0033) to the regression model significantly increased the ability to predict the proximal medial lateral bone distribution (R2=0.72, p=0.0001). Distally neither the subject characteristics nor the gait moments and predicted forces were significant predictors of the bone distribution. The lack of a correlation distally may be reflective of the forces being more evenly distributed further from the tibial plateau. While it has long been suggested that the adduction moment is the primary determinate of the distribution of load between the medial and lateral plateaus, this is the first evidence of its relationship to the underlying bone distribution. PMID- 9727340 TI - Backside nonconformity and locking restraints affect liner/shell load transfer mechanisms and relative motion in modular acetabular components for total hip replacement. AB - Nonconformity between the polyethylene liner and the metal shell may exist in modular acetabular components by design, due to manufacturing tolerances, or from locking mechanisms that attach the polyethylene liner to the metal shell. Relative motion at the liner/shell interface has been associated with backside wear, which may contribute to osteolysis which has been clinically observed near screw holes. The purpose of this study was to investigate the effect of nonconformity and locking restraints on the liner/shell relative motion and load transfer mechanisms in a commercially available, metal-backed acetabular component with a polar fenestration. The finite element method was used to explore the hypothesis that backside nonconformity and locking restraints play important roles in long-term surface damage mechanisms that are unique to modular components, such as backside wear and liner extrusion through screw holes. The three-body quasi-static contact problem was solved using a commercially available explicit finite element code, which modeled contact between the femoral head, polyethylene liner, and the metal shell. Four sets of liner boundary conditions were investigated: no restraints, rim restraints, equatorial restraints, and both rim and equatorial restraints. The finite element model with a conforming shell predicted between 8.5 and 12.8 microm of incremental extrusion of the polyethylene through the polar fenestration, consistent with in vitro experiments of the same design under identical loading conditions. Furthermore, idealized rim and/or equatorial liner restraints were found to share up to 71% of the load across the liner/shell interface. Consequently, the results of this study demonstrate that backside nonconformity and locking restraints substantially influence backside relative motion as well as load transfer at the liner/shell interface. PMID- 9727341 TI - A comparative study of impact dynamics: wobbling mass model versus rigid body models. AB - We demonstrate that the results of inverse dynamics depend sensitively on the model used to simulate the human body. For this reason we produce "ideal" input data of the ground reaction force and the heel acceleration for a computer simulated drop jump with a wobbling mass model. Then we determine the internal torques and forces in the knee and hip joint via inverse dynamics by employing a rigid body model. It turns out that during the impact phase the analysis with the rigid body model yields completely incorrect internal torques and forces. PMID- 9727342 TI - Incompressibility of the solid matrix of articular cartilage under high hydrostatic pressures. AB - The objective of this study was to test the hypothesis that the organic solid matrix of articular cartilage is incompressible under physiological levels of pressure. Due to its anisotropic swelling behavior, an anisotropic version of the biphasic theory was used to predict the deformation and internal stress fields. This theory predicts that, under hydrostatic loading of cartilage via a pressurized external fluid, a state of uniform hydrostatic fluid pressure exists within the tissue regardless of the anisotropic nature of the solid matrix. The theory also predicts that if the solid matrix is intrinsically incompressible, the tissue will not deform under hydrostatic loading conditions. This prediction, i.e., no deformation, was experimentally tested by subjecting specimens of normal bovine articular cartilage to hydrostatic pressures. A new high pressure hydrostatic loading chamber was designed and built for this purpose. It was found that normal bovine articular cartilage, when subject to hydrostatic pressures up to 12 M Pa, does not deform measurably. This experimental finding supports one of the fundamental assumptions of the biphasic theory for cartilage, i.e., the organic solid matrix of the tissue is intrinsically incompressible when loaded within the normal physiologic range of pressures. Hydrostatic loading has often heen used in cartilage explant cultures for tissue metabolism studies. The findings of this study provides an accurate method to calculate the states of stress acting on the fluid and solid phases of the tissue in these hydrostatically loaded explant culture experiments, and suggest that tissue deformation will be minimal under pure hydrostatic pressurization. PMID- 9727343 TI - Analysis of 3D transient blood flow passing through an artificial aortic valve by Lattice-Boltzmann methods. AB - The development of flow instabilities due to high Reynolds number flow in artificial heart valve geometries inducing high strain rates and stresses often leads to hemolysis and related highly undesired effects. Geometric and functional optimization of artificial heart valves is therefore mandatory. In addition to experimental work in this field it is meanwhile possible to obtain increasing insight into flow dynamics by computer simulation of refined model problems. After giving an introductory overview we report the results of the simulation of three-dimensional transient physiological flows in fixed geometries similar to a CarboMedics bileaflet heart valve at different opening angles. The visualization of emerging complicated flow patterns gives detailed information about the transient history of the systems dynamical stability. Stress analysis indicates temporal shear stress peaks even far away from walls. The mathematical approach used is the Lattice Boltzmann method. We obtained reasonable results for velocity and shear stress fields. The code is implemented on parallel hardware in order to decrease computation time. Finally, we discuss problems, shortcomings and possible extensions of our approach. PMID- 9727344 TI - A model equation for the prediction of mechanical internal work of terrestrial locomotion. AB - By refining a previously published model, a simple equation for the estimation of the mechanical internal work during locomotion is presented. The only input variables are the progression speed, the stride frequency and the duty factor, i.e. the fraction of the stride duration at which a foot is in contact with the ground. The inclusion of this last variable, easily measurable, allows to obtain a single equation for both walking and running. The model predictions have been compared with the mechanical internal work experimentally obtained on humans in several conditions: speeds (range 0.8-3.3 m s(-1)), gaits (walking and running) and gradients (+/-15%). The close match between the two indicates that the model equation can be used whenever a direct measurement of the mechanical internal work is unavailable. PMID- 9727345 TI - Force feedback control of motor unit recruitment in isometric muscle. AB - The use of simple force feedback in an isometric muscle control system utilizing orderly recruitment of motor units is explored. Cat medial gastrocnemius motor units were stimulated with and without simple force feedback gain ranging from 0.7 to 0.9. Ramp, triangular, staircase, sinusoidal and bandwidth-limited pseudo random input recruitment signals were used to study tracking accuracy through linear correlation in ramp and triangular signals, cross correlation in sinusoidal and random signals, and rise time and steady state error in staircase signals. Dramatic improvements were found in most tested tracking variables with the use of feedback; squared correlation coefficients increased from a mean of 0.93 to 0.99 for ramp signals and from 0.76 to 0.98 in triangular signals. Mean peak cross-correlations improved from 0.85 to 0.98 in random signals and from 0.93 to 0.98 for sinusoidal inputs, and mean time to peak cross-correlations decreased from 144 to 24 ms in random signals and from 156 to 25 ms in sine waves. Rise times in staircase signals decreased from a mean of 520 to 175 ms, and mean steady state error decreased from 12 to 3%. Significant effects of the triangle cycle time, sinusoidal frequency and staircase step were found on the performance of the muscle force control system. In addition, the possible effects of intrinsic feedback mechanisms on the control system were examined by repeating the closed loop part of the study but with the sciatic nerve cut proximally. The tracking results were essentially and statistically the same as in the closed loop condition. It was concluded that a simple feedback configuration provided superior tracking performance for a practical application in which orderly recruitment is used to control muscles; furthermore, it was concluded that this type of system would be virtually immune to external disturbances such as spasticity resulting from intact spinal neural feedback mechanisms found in paralyzed individuals. PMID- 9727346 TI - Dynamics of coronary artery curvature obtained from biplane cineangiograms. AB - A system was developed to track the centerlines (axes) of human coronary arteries during the cardiac cycle from biplane cineangiograms. The system employs a time interpolation technique on one image plane and a smoothing technique on the other to create simultaneous image pairs. The image pairs are then used to reconstruct the three dimensional axes of the vessels during the cardiac cycle. Once the vessel axes are reconstructed, acceleration, velocity and geometric parameters such as curvature can be obtained. In the present work, a sequence of human angiograms is used to obtain the axial and temporal variation of the curvature of a portion of the left anterior descending coronary artery distal to the third diagonal branch during two cardiac cycles. For the case studied, the change in curvature during the cycle ranged from 0.25 to 1.8 cm(-1), depending upon the position of the site. PMID- 9727347 TI - Mechanical properties of the tendinous equine interosseus muscle are affected by in vivo transducer implantation. AB - Liquid metal strain gauges (LMSGs) were implanted in the tendinous interosseous muscle, also called suspensory ligament (SL), in the forelimbs of 6 ponies in order to quantify in vivo strains and forces. Kinematics and ground reaction forces were recorded simultaneously with LMSG signals at the walk and the trot prior to implantation, and 3 and 4 days thereafter. The ponies were euthanised and tensile and failure tests were performed on the instrumented tendons and on the tendons of the contra lateral limb, which were instrumented post mortem. The origo-insertional (OI) strain of the SL was computed from pre- and post-operative kinematics, using a 2D geometrical model. The LMSG-recorded peak strain of the SL was 5.4+/-0.9% at the walk and 9.1+/-1.3% at the trot. Failure occurred at 15.4+/ 2.1% (mean+/-S.D.). The LMSG strain was higher than the simultaneously recorded OI strain 0.5+/-0.7% strain at the walk and 2.2+/-1.1% strain at the trot. Post operative OI strains were only slightly higher than pre-operative values. Failure strains of in vivo instrumented SLs were 2.0+/-1.2% strain higher, and failure forces were slightly lower, than those of the contra lateral SLs that were instrumented post mortem. SL strains appeared to be considerably higher than those found in earlier acute experiments. Differences between in vivo LMSG and OI strains, supported by lower failure strains comparing in vivo and post mortem instrumented SLs, revealed that local changes in tendon mechanical properties occurred within 3 to 4 days after transducer implantation. Therefore, measurements of normal physiological tendon strains should be performed as soon as possible after transducer implantation. PMID- 9727348 TI - Fast and accurate automated measurements in digitized stereophotogrammetric radiographs. AB - Until recently, Roentgen Stereophotogrammetric Analysis (RSA) required the manual definition of all markers using a high-resolution measurement table. To automate this tedious and time-consuming process and to eliminate observer variabilities, an analytical software package has been developed and validated for the detection, identification, and matching of markers in RSA radiographs. The digital analysis procedure consisted of the following steps: (1) the detection of markers using a variant of the Hough circle-finder technique; (2) the identification and labeling of the detected markers; (3) the reconstruction of the three-dimensional position of the bone markers and the prosthetic markers; and (4) the computation of micromotion. To assess the influence of film digitization, the measurements obtained from nine phantom radiographs using two different film scanners were compared with the results obtained by manual processing. All markers in the phantom radiographs were automatically detected and correctly labeled. The best results were obtained with a Vidar VXR-12 CCD scanner, for which the measurement errors were comparable to the errors associated with the manual approach. To assess the in vivo reproducibility, 30 patient radiographs were analyzed twice with the manual as well as with the automated procedure. Approximately, 85% of all calibration markers and bone markers were automatically detected and correctly matched. The calibration errors and the rigid-body errors show that the accuracy of the automated procedure is comparable to the accuracy of the manual procedure. The rigid-body errors had comparable mean values for both techniques: 0.05 mm for the tibia and 0.06 mm for the prosthesis. The reproducibility of the automated procedure showed to be slightly better than that of the manual procedure. The maximum errors in the computed translation and rotation of the tibial component were 0.11 mm and 0.24, compared to 0.13 mm and 0.27 for the manual RSA procedure. The total processing time is less than 10 min per radiograph, including interactive corrections, compared to approximately 1 h for the manual approach. In conclusion, a new and widely applicable, computer-assisted technique has become available to detect, identify, and match markers in RSA radiographs and to assess the micromotion of endoprostheses. This new technique will be used in our clinic for our hip, knee, and elbow studies. PMID- 9727349 TI - Eccentric training does not increase sarcomere number in rabbit dorsiflexor muscles. AB - The hypothesis of this study was that eccentric muscle training induces serial sarcomere addition. Eccentric training of rabbit dorsiflexor muscles (50 maximal contractions, 2 times per week, for 12 weeks) increased sarcomere number by only 3% in superficial fascicles of the tibialis anterior (TA) and did not change sarcomere number in deep fascicles of the TA nor in proximal or distal fascicles of extensor digitorum longus. Thus eccentric training had little or no effect on sarcomere number. These results do not support previous speculation that eccentric training produces increases in sarcomere number. PMID- 9727350 TI - Neural induction in embryos. AB - Neural differentiation of the ectoderm is inhibited by bone morphogenetic protein 4 (BMP-4) in amphibia as well as mammalia. This inhibition is released by neural inducing factor(s), which are secreted from the dorsal mesoderm. Masked neuralizing factor(s) are already present in the ectoderm before induction. In homogenates from Xenopus oocytes and embryos neural inducing factors were found in the supernatant (centrifuged at 105000 g), in small vesicles and a ribonucleoprotein fraction. A neuralizing factor, which is a protein of small size, has been partially purified from Xenopus gastrulae. Genes that are expressed in the dorsal mesoderm and involved in the de novo synthesis of neuralizing factor(s) have been cloned. The differentiation of cells with a neuronal fate starts in the neural plate immediately after neural induction. Genes homologous to the Notch and Delta genes of lateral inhibition in insects are involved in this process. PMID- 9727351 TI - DNA replication cycle in parthenogenetically developing eggs of the starfish Asterina pectinifera. AB - Starfish oocytes artificially activated by a calcium ionophore will develop normally if the formation of polar bodies is suppressed. In the present paper, schedules of the DNA replication period (S phase) of these parthenogenotes were explicitly timed using 5-bromo-2'-deoxyuridine (BrdU) and anti-BrdU monoclonal antibody. Their schedule of S phase was identical to that of fertilized eggs. Consequently, an S phase regulation system is triggered even in parthenogenotes raised by dual treatment of egg activation and polar body suppression. The S phase schedule of parthenogenotes confirms the temporal pattern of chromosome duplication, observed by other researchers, leading to tetraploid parthenogenotes. The S phase determination also provides a basis for argument concerning the number of centrioles participating in parthenogenetic development. If polar body formation of activated eggs was not suppressed, the first S phase was normal, but the second S phase did not recur on time. A rigidly regulated system of DNA replication cycle, which should be an essential prerequisite for parthenogenesis, thus requires the content of polar bodies. PMID- 9727352 TI - Developmental regulation and tissue-specific localization of calmodulin mRNA in the protochordate Ciona intestinalis. AB - A full-length cDNA encoding a highly conserved calmodulin was isolated from a cDNA library prepared from hatched larvae of the ascidian Ciona intestinalis. Sequence analysis has identified a 447 b.p. open reading frame, encoding a putative protein of 149 amino acid residues, with a predicted molecular weight of 16.8 kDa, showing 85-98% identity to known calmodulins. Northern blot analysis revealed a single transcript of about 0.8 kb in length, which was maternally expressed and progressively increased during development, until late tail-bud stage. Whole-mount in situ hybridizations, carried out on embryos at different stages of development, showed that starting from the neurula stage, the C. intestinalis calmodulin (Ci-CaM) expression became restricted to the neuroectoderm and that in larvae it was specifically detected in the nervous system. PMID- 9727353 TI - Mapping of cholecystokinin transcription in transgenic mouse brain using Escherichia coli beta-galactosidase reporter gene. AB - Cholecystokinin (CCK), a neuro-gut peptide, occurs not only in the nervous but also in the digestive system. As a first step in elucidating whether CCK gene expression and its physiological functions co-operate in these separate organs, transgenic mice were produced using CCK promoter that directs bacterial beta galactosidase as a reporter gene. A new transgenic vector was constructed, inserting the SV40 poly A signal 5' to the CCK promoter to impede any transcription upstream of the transgene. A 2.4 kb.p. region upstream to the transcription start site of the mouse CCK gene was used as the promoter. Transgene expression was detected first at embryonic 13.5 days in the central nervous system and increased after birth. The distribution of cells expressing beta-galactosidase transgene agreed fairly well with that of in situ hybridization. In addition, the upregulation of CCK gene expression was clearly demonstrated by expressing beta-galactosidase after injury to the brain. These results indicated that the 2.4 kb.p. of the CCK gene promoter region was sufficient to direct appropriate tissue-specific gene expression in mice. PMID- 9727354 TI - Juxtaposition of two heterotypic monolayer cell cultures of chick limb buds. AB - In chick limb buds, mesenchymal cells of the progress zone (PZ-cells) at different developmental stages segregate one from the other in mixed cell cultures, suggesting they have different cell affinity. In order to learn the possible roles of such differences in the cells, two heterotypic leg PZ-cell populations (cells from stages 25/26 and 20/21) in vitro were juxtaposed to allow them to form the boundary. A method with double cylindrical columns was used to make adjoining monolayer cell cultures. It was shown that heterotypic juxtaposition produced two chondrogenic patterns along the boundary: aggregates of chondrocytes formed by stage 20/21 PZ-cells and a chondrocyte-free band formed by those at stage 25/26. Juxtaposition of PZ-cells and proximal cells also formed these patterns, while that between cells from anterior and posterior PZ formed indistinct patterns along the boundary. Homotypic PZ-cell juxtaposition did not produce these patterns. The results suggest that different cell affinity has a role in the segmentation of cartilage patterns at a point along the proximodistal axis, as well as a role in retaining cells in one area so as not to be recruited to other condensation areas. PMID- 9727355 TI - Retinoic acid affects patterning along the anterior-posterior axis of the ascidian embryo. AB - Because retinoic acid (RA) is known to affect anterior posterior patterning in vertebrate embryos, it was questioned whether it shows similar effects in a more primitive chordate, the ascidian Halocynthia roretzi. Ascidian embryos treated with RA exhibited truncated phenotypes in a dose-dependent manner similar to the anterior truncations seen in vertebrate embryos. The most severely affected larvae possessed a round trunk without the papillae characteristic of the anterior terminal epidermis. Retinoic acid also altered the expression of HrHox-1 and Hroth in a dose-dependent manner. Expression of HrHox-1 increased, whereas expression of Hroth decreased with increasing levels of RA. In treated embryos, HrHox-1 was first expressed pan-ectodermally, then degraded in all but specific regions of the embryo. By contrast, initiation of Hroth expression was not affected, but epidermal expression was lost while expression in the neural tube narrowed toward the anterior in tail-bud embryos. These alterations in the expression of homeobox genes appear to correlate closely to the morphological defects elicited by RA treatment, suggesting broad conservation of developmental patterning mechanisms within the Phylum Chordata. PMID- 9727356 TI - Apical ectodermal ridge induction by the transplantation of En-1-overexpressing ectoderm in chick limb bud. AB - In the early chick embryo, the dorsal ventral (DV) boundary organizes the apical ectodermal ridge (AER) structure in the limb bud field. Here it is reported that Engrailed-1 (En-1), a homolog of the Drosophila segment polarity gene engrailed expressed in the ventral limb ectoderm, participates in AER formation at the DV boundary of the limb bud. Restricted ectopic expression of En-1 in the dorsal side of the limb bud by transplantation of En-1-overexpressing ectoderm induces ectopic AER at the boundary of En-1-positive and -negative cells. The results suggest that En-1 is involved in AER formation at the DV boundary of the limb bud. PMID- 9727357 TI - An ascidian gene encoding an SH2-domain protein is expressed in the notochord cells of the embryo. AB - Differentiation of notochord cells in the ascidian embryo requires cell-cell interactions and signal transduction pathways. Isolation and characterization of an ascidian gene (HrSH2) from Halocynthia roretzi is reported. Sequence analysis suggests that HrSH2 encodes a polypeptide with an SH2 domain and a tyrosine kinase phosphorylation site, that are implicated in signaling pathways through tyrosine phosphorylation. Zygotic expression of HrSH2 was transient. The gene expression began at the 110-cell stage but was downregulated by the larval stage. Whole-mount in situ hybridization, taking advantage of well-known lineage, revealed that the HrSH2 transcript first appeared in primordial notochord cells as well as a few endoderm cells of the 110-cell embryo. During gastrulation and neurulation, expression in the endoderm was downregulated, and instead HrSH2 transcript became evident in notochord cells, nerve cord cells, endodermal strand cells and epidermal cells of the tail. These results suggest the possibility that HrSH2 is involved in the signal transduction pathways required for notochord formation and for differentiation of other cells in conjunction with the notochord. PMID- 9727358 TI - Timing and mechanisms of mesodermal and neural determination revealed by secondary embryo formation in Cynops and Xenopus. AB - We examined the timing and mechanisms of mesodermal and neural determination in Cynops, using the secondary embryo induced by transplantation of the prechordal endomesoderm. Two unique approaches were used: one was to observe gastrulation movements induced by the graft, and the other to measure the volumes of formed tissues. Transplanted graft pulled host animal cap cells inside to form a new notochord and other mesoderm of the secondary embryo, showing determination of mesoderm during gastrulation. The graft attained a certain width beneath the host ectoderm and moved near to the animal pole of the host by late gastrula, and a neural plate, which had a similar width to the graft, was formed covering the graft. The volume of neural tissues of the secondary embryo at tail-bud stages was about half that of the normal embryo, while the volumes of notochord were comparable in each case. These data suggest that prechordal endomesoderm, rather than notochord, determines the limit of neural plate in the overlying ectoderm. Similar dorsal grafts were transplanted at early gastrula in Xenopus but did not form well developed secondary embryos, demonstrating that the timing and mechanisms of mesoderm formation in Xenopus are different from those in Cynops. PMID- 9727359 TI - Improved hatching for in vitro quail embryo culture using surrogate eggshell and artificial vessel. AB - The establishment of avian embryonic culture is important both for the analysis of the developmental process and the establishment of transgenic chickens that produce useful biological materials in eggs. However, the hatchability of cultured embryos has been approximately 50%. We identified that the low rate of hatchability of cultured embryos was caused by limited oxygen and calcium availability. In quail embryo culture using chicken eggshell as a culture vessel, viability in the middle stage of culture was improved and 30% of embryos were hatched by oxygen enrichment. Furthermore, hatchability increased to 80% by supplementation with calcium lactate in addition to oxygen aeration. In the present study, a fully artificial vessel for quail embryo culture was designed using a gas-permeable Teflon membrane. By the addition of fine eggshell powder and calcium lactate, quail embryos grew and developed normally, and 43% of embryos hatched. Although the hatchability was lower than that of cultures using a surrogate eggshell, we achieved in hatching an avian embryo using a fully artificial vessel. PMID- 9727360 TI - Shh, Bmp-2 and Hoxd-13 gene expression in chick limb bud cells in culture. AB - The anteroposterior axis of the vertebrate limb bud is determined by signals from the zone of polarizing activity (ZPA). Sonic hedgehog (Shh) is expressed in the posterior mesoderm, which corresponds closely to ZPA activity. Moreover, Bmp-2 and HoxD genes are expressed in the broader posterior mesoderm, and it is thought that the ZPA signaling pathway consists of these gene products. Limb outgrowth and patterning, including expression of these genes, depend on the apical ectodermal ridge (AER). Fibroblast growth factors (FGF) have been identified as candidates for signal molecules from the AER. To further understand the ZPA signaling pathway and the participation of FGF, expressions of these genes were examined by reverse transcription-polymerase chain reaction in chick limb bud cells cultured with FGF-4. The present results indicate that FGF-4 cannot maintain Shh expression but can maintain Hoxd-13 expression in cultured posterior cells; moreover, Bmp-2 is expressed independently of FGF-4. These results suggest that Bmp-2 and Hoxd-13 expressions do not require a continuous expression of Shh. Further, it was demonstrated that posterior cells cultured with FGF-4 recovered Shh expression when grafted to the limb bud, indicating that FGF-4 maintains not Shh expression itself but competence of Shh expression. PMID- 9727361 TI - Glycosylation is not essential for vasopressin-dependent routing of aquaporin-2 in transfected Madin-Darby canine kidney cells. AB - Glycosylation has been shown to be important for proper routing and membrane insertion of a number of proteins. In the collecting duct, aquaporin-2 (AQP2) is inserted into the apical membrane after stimulation of vasopressin type-2 receptors and retrieved into an endosomal compartment after withdrawal of vasopressin. The extent of glycosylation of AQP2 in human kidney and urine and the effects of deglycoylation on routing of AQP2 in an AQP2-transfected Madin Darby canine kidney cell line (clone WT10) were investigated. Semiquantitative immunoblotting of human kidney membranes and urine showed an AQP2 glycosylation of 35 to 45% for medulla, papilla, and urine, with low variation among individuals. The 1-desamino-8-D-arginine vasopressin-induced transcellular osmotic water permeability (Pf) of WT10 cells by a factor of 2.6 +/- 0.2 was reduced to 1.5 +/- 0.1 after pretreatment with the glycosylation inhibitor tunicamycin. However, when WT10 cells were incubated with 8-br-cAMP, the Pf increased by a factor 2.8 +/- 0.2 and by 2.9 +/- 0.2 after prior incubation with tunicamycin. Immunoblot analyses revealed that in WT10 cells, 34% of AQP2 is glycosylated, which was reduced to 2% after tunicamycin treatment. Surface biotinylation and subsequent semiquantitative immunoblotting revealed that stimulation by cAMP increased the level of AQP2 in the apical membrane of WT10 cells 1.5-fold. independent of the presence of tunicamycin. However, in tunicamycin-treated WT10 cells, all AQP2 in the apical membrane was unglycosylated, whereas in untreated cells 30% of AQP2 in the apical membrane was glycosylated. These results prove that glycosylation has no function in the routing of AQP2 in Madin-Darby canine kidney cells. PMID- 9727363 TI - Dietary sulfate regulates the expression of the renal brush border Na/Si cotransporter NaSi-1. AB - Dietary inorganic sulfate (Si) intake is an important factor in the regulation of renal proximal tubular sodium-dependent Si transport (Na/Si cotransport). The purpose of the present study was to determine whether modulation of Na/Si cotransport activity by dietary Si is mediated through regulation of the renal expression of the recently cloned NaSi-1 protein located in the apical brush border membrane (BBM) of the proximal tubule. It was found that rats fed a high Si diet had a marked increase in the renal excretion of Si and a concomitant decrease in BBM Na/Si cotransport activity when compared with rats on a control Si diet. The 43% decrease in BBM Na/Si cotransport activity was associated with a 33% decrease in BBM NaSi-1 protein abundance, as determined by Western blotting, and a 2.7-fold decrease in cortical NaSi-1 mRNA abundance, as determined by Northern blotting. Furthermore, cortical mRNA from rats fed a high Si diet when injected into Xenopus laevis oocytes led to a 2.2-fold decrease in Na/Si cotransport activity compared with mRNA isolated from control Si diet rats. This study indicates that adaptation to a high Si diet is accompanied by a decrease in renal cortical NaSi-1 mRNA abundance, which results in reduced expression of the NaSi-1 protein at the level of the proximal tubular BBM. PMID- 9727362 TI - Renal Na/H exchanger NHE-3 and Na-PO4 cotransporter NaPi-2 protein expression in glucocorticoid excess and deficient states. AB - Administration of pharmacologic doses of glucocorticoid in vivo increases renal proximal tubule apical membrane Na/H exchange and decreases Na/PO4 cotransport activity (1). Current data suggest that the NHE-3 and NaPi-2 proteins mediate significant fractions of proximal tubule apical membrane Na/H exchange and Na/PO4 cotransport, respectively. This study examines whether glucocorticoid excess or deficiency affects NHE-3 and NaPi-2 protein abundance and the intrarenal distribution of these transporters. Protein abundance of NHE-3 and NaPi-2 in control rats was compared to rats rendered glucocorticoid-deficient by bilateral adrenalectomy, and to rats receiving pharmacologic doses of dexamethasone using immunoblots and immunohistochemistry. Adrenalectomy had modest effects on NHE-3 protein abundance, but dexamethasone administration to either adrenalectomized or sham-operated rats significantly increased NHE-3 protein abundance in both the proximal tubule and thick ascending limb, but not the thin descending limb. Adrenalectomy increased NaPi-2 protein abundance in the proximal tubule, whereas dexamethasone administration dramatically suppressed NaPi-2 protein on the apical membrane in both adrenalectomized and sham-operated animals. No significant reciprocal increase in subapical NaPi-2 staining was seen in the dexamethasone treated rats. The present study shows that glucocorticoids regulate proximal tubule apical membrane Na/H exchange and NaPi cotransport by changes in protein abundance of NHE-3 and NaPi-2, respectively. PMID- 9727364 TI - Function and expression of a novel rat salt-tolerant protein: evidence of a role in cellular sodium metabolism. AB - Higher dietary salt intake in humans is associated with higher BP, but the BP response to NaCl, so-called salt sensitivity, is heterogeneous among individuals. It has been postulated that modifications in cellular cation metabolism may be related to salt sensitivity in mammalian hypertension. The authors have isolated a novel rat complementary DNA, called salt-tolerant protein (STP), that can functionally complement Saccharomyces cervisiae HAL1, which improves salt tolerance by modulating the cation transport system. On high-salt (8% NaCl) diets, both Dahl salt-sensitive and salt-resistant rats displayed an elevated BP and increased STP mRNA expression. Immunohistochemistry using an anti-rat STP antibody demonstrated the presence of STP immunoreactivity in the proximal tubules. In cells that transiently expressed STP, the intracellular [Na+]/[K+] ratio was higher than that in control cells. STP contains predicted coiled-coil and Src homology 3 domains, and shows a partially high degree of nucleotide identity to human thyroid-hormone receptor interacting protein. These results suggest that STP may play an important role in salt sensitivity through cellular sodium metabolism by mediating signal transduction and a hormone-dependent transcription mechanism. PMID- 9727365 TI - Evidence against elevated sympathetic vasoconstrictor activity in borderline hypertension. AB - The relationship between sympathetic nerve activity and BP in the early stages of essential hypertension remains unclear. To investigate this relationship further, this study measured resting muscle sympathetic nerve activity (MSNA: representing peripheral vasoconstrictor activity), plasma catecholamines, BP, central venous pressure, and heart rate in 20 young (24 +/- 2 SD yr), lean (body mass index, 24.2 +/- 3.0 kg/m2), male subjects with borderline hypertension (BHT) and in 21 male normotensive (NT) control subjects matched for age and body mass index. A cold pressor test was also performed to evaluate sympathetic reflex responsiveness. Resting mean BP and heart rate were significantly higher in the BHT subjects compared with NT subjects (113 +/- 9 versus 89 +/- 9 mmHg; 74 +/- 8 versus 62 +/- 8 bpm; P < 0.0001 each) with no difference in central venous pressure. Resting MSNA levels tended to be lower in the BHT versus the NT group (12 +/- 6 versus 14 +/- 9 bursts/min, P = NS; 16 +/- 8 versus 22 +/- 13 bursts/100 heartbeats, P = 0.05) and did not correlate with either BP or body mass index. Significant positive correlations were found between resting MSNA and plasma norepinephrine levels in both groups (P < 0.05). Hemodynamic and sympathetic nerve responses to the cold pressor test were similar between the BHT and NT subjects. It is concluded that resting MSNA and plasma norepinephrine levels are correlated in young lean NT and BHT men; however, neither of these variables is correlated with BP. Because MSNA was similar in the two groups, the concept that augmented resting MSNA is important in the early developmental phase of essential hypertension must be reevaluated. PMID- 9727366 TI - Interactions between nitric oxide and renal nerves on pressure-diuresis and natriuresis. AB - The present study examined the effect of renal denervation on the impairment of the pressure-diuresis response produced by nitric oxide synthesis blockade. The experiments were performed in Inactin-anesthetized Munich-Wistar rats. The animals with innervated kidneys had lower baseline values of renal blood flow, GFR, sodium excretion (UNaV), and urine flow (V) than rats with denervated kidneys. Also, renal denervation shifted pressure-diuresis and natriuresis toward lower pressures. A low dose of N(omega)-nitro-L-arginine methyl esther (NAME, 3.7 nmol/kg per min) reduced UNaV and the fractional excretion of sodium (FENa) and blunted pressure-natriuresis only in rats with innervated kidneys, whereas it had no effects in rats with denervated kidneys. A medium dose of NAME (37 nmol/kg per min) lowered FENa only in rats with innervated kidneys. The administration of NAME (37 nmol/kg per min) blunted pressure-diuresis and natriuresis in kidneys with or without the renal nerves, but the effect was more pronounced in rats with innervated kidneys. A high dose of NAME (3.7 micromol + 185 nmol/kg per min) increased UNaV and FENa only in rats with innervated kidneys, whereas it reduced GFR, V, UnaV, and FENa in rats with denervated kidneys. However, pressure natriuresis and diuresis were blunted by this high dose of NAME independently of the presence or absence of renal nerves. These results demonstrate that renal nerves potentiate the renal effects of low doses of NAME on renal function and pressure-diuresis and natriuresis. However, high doses of NAME abolish pressure diuresis independently of renal nerves, and the natriuretic effect of NAME in innervated kidneys may be attributed to reflex inhibition of sympathetic tone due to the rise in arterial pressure. PMID- 9727367 TI - Nitric oxide release as an essential mitigating step in tubuloglomerular feedback: observations during intrarenal nitric oxide clamp. AB - Nitric oxide synthase inhibition in the kidney enhances tubuloglomerular feedback (TGF) responsiveness. This may reflect either the effect of reduced basal nitric oxide (NO) availability or the effect of impaired NO release that is physiologically induced by TGF activation. However, it is unknown whether the latter actually takes place. In this study, it was hypothesized that NO is released (from macula densa cells or endothelium) as part of the normal TGF loop, and mitigates the TGF response. In Sprague Dawley rats, TGF responsiveness was assessed (fall in tubular stop flow pressure, deltaSFP, upon switching loop of Henle perfusion rates from 0 to 40 nl/min) during an intrarenal NO clamp (systemic infusion of nitro-L-arginine, 10 microg/kg per min, followed by intrarenal nitroprusside infusion adjusted to restore renal blood flow [RBF]). This maneuver was presumed to fix intrarenal NO impact at a physiologic level. To validate the approach, TGF responsiveness during an intrarenal angiotensin II (AngII) clamp (systemic infusion of enalaprilat 0.2 mg/kg per min, followed by intrarenal AngII infusion) was also studied. AngII is presumed to modulate but not mediate, TGF, thus not to increase as part of the TGF loop. In untreated animals, RBF was 7.4 +/- 0.4 ml/min, and deltaSFP was 5.7 +/- 1.6 mmHg. Nitro-L arginine infusion alone reduced RBF to 5.3 +/- 0.5 ml/min (P < 0.05); with nitroprusside infusion, RBF was restored to 8.3 +/- 0.7 ml/min. In this condition (NO clamp), deltaSFP was markedly increased to 19.6 +/- 3.2 mmHg (P < 0.05). By contrast, deltaSFP, which was virtually abolished during enalaprilat alone (0.2 +/- 0.3 mmHg), was not significantly different from controls during AngII clamp (8.2 +/- 1.0 mmHg). These data suggest that NO may well be released upon TGF activation. By contrast, AngII is not dynamically involved in TGF activation, but may modulate the TGF response. Thus, dynamic release of NO during TGF activation mitigates the TGF response, so that it will offset the action of a primary, as yet undefined, vasoconstrictor mediator. The source of this NO, macula densa or endothelium, remains to be elucidated. PMID- 9727368 TI - Does dopamine use several signal pathways to inhibit Na-Pi transport in OK cells? AB - In opossum kidney (OK) cells, L-dihydroxyphenylalanine (10 microM) raised dopamine to 10 nM and inhibited Na-inorganic phosphate (Pi) uptake 20% (P = 0.001). Inhibition was completely blocked by carbidopa or SCH23390. Dopamine (1 microM) inhibited uptake 55% (half-maximal inhibition, 0.03 microM). Fenoldopam (0.1 microM, DA1 agonist) inhibited uptake 45 +/- 2%. DA1 antagonists (SKF83566 and SCH23390), but not DA2-antagonist (sulpiride), blocked dopamine inhibition. Quinpirole (DA2 agonist) did not modify Pi uptake. Bisindolylmaleimide (10 microM), a protein kinase C inhibitor, blocked inhibition of Pi uptake by phorbol ester but had no effect on the response to dopamine. Dopamine inhibited Pi uptake in cells that had been exposed to phorbol ester for 18 to 24 h. Dopamine inhibition was not reduced by 1 microM U73,122 but was reduced 20% by 10 microM, which is 10 times the concentration reported to completely inhibit phospholipase C in OK cells. Adenylate cyclase inhibitors SQ 22536 (100 microM) and 2,5 dideoxyadenosine (100 microM) reduced dopamine-stimulated cAMP production, but not dopamine inhibition of Pi uptake. Rp-cAMPS counteracted the inhibition of Pi uptake by Sp-cAMPS but had no effect on the dopamine response. H-89 inhibited dopamine-stimulated protein kinase A activity, but neither H-89 nor H-9 alone or with bisindolylmaleimide altered dopamine inhibition of Pi uptake. Genistein and herbimycin A (tyrosine kinase inhibitors) reduced Pi uptake. However, dopamine, a benzoquinone like several tyrosine kinase inhibitors, did not inhibit tyrosine kinase activity. Thus, dopamine inhibited Pi uptake in this OK cell clone by activating a G protein-linked pathway that operates independently from adenylyl cyclase, protein kinase A, protein kinase C, and protein tyrosine kinase. PMID- 9727369 TI - Upregulation of atrial natriuretic peptide gene expression in remnant kidney of rats with reduced renal mass. AB - Atrial natriuretic peptide (ANP) is synthesized in the kidney but its physiologic significance there is unclear. To determine whether renal expression of the ANP gene is regulated, renal ANP mRNA expression was assessed in remnant kidneys after 5/6 nephrectomy in Munich-Wistar rats. In normal sodium intake groups, ANP mRNA expression in the remnant kidney was significantly increased by 5.0 +/- 0.8 fold (n = 7, mean +/- SEM) at 4 d when compared with sham-operated controls (n = 6, all sham-operated groups) (*P < 0.001 by Scheffe's test) and by 28.3 +/- 5.1 fold at 14 d. This latter response was markedly diminished to 7.6 +/- 2.1-fold (n = 7, versus sham) in rats maintained on a low sodium diet. At 4 d, on the other hand, no significant downregulation was observed with dietary sodium restriction. Because natriuretic peptides have previously been shown by us to play a major role in the adaptive responses of remnant nephrons to renal mass ablation, these data suggest that ANP of renal origin may contribute to the overall mechanism for enhancing sodium excretion in the face of declining nephron number. PMID- 9727370 TI - Regulation of nuclear factor kappaB by corticosteroids in rat mesangial cells. AB - Nuclear factor kappaB (NF-kappaB) is one of the most important proinflammatory transcription factors. The anti-inflammatory activity of steroids in leukocytes is partly due to inhibition of signaling by NF-kappaB, but it is not known whether steroids inhibit NF-kappaB in kidney cells. Since the mesangial cell is important in several kidney diseases, especially mesangial proliferative glomerulonephritis, the aims of this study were: (1) to define the mechanism of NF-kappaB activation in rat glomerular mesangial cells; and (2) to determine whether steroids inhibit activation of NF-kappaB in these cells. Electrophoretic mobility shift assays (EMSA) showed that interleukin-1beta and tumor necrosis factor-alpha activated NF-kappaB from 15 min to 48 h after stimulation. Supershift EMSA demonstrated that p65 and p50 were the predominant subunits involved. Degradation of the inhibitory subunit IkappaB-alpha was first observed 15 min after stimulation by Western blot, was maximal at 15 to 30 min (>90% by densitometry), and had returned to near normal levels at 90 min. In contrast, IkappaB-beta was maximally degraded at 60 to 120 min and was still reduced at 48 h (<50% of the untreated level). Although treatment of mesangial cells with dexamethasone increased IkappaB-alpha mRNA by 1.92x and protein by 1.45x over controls, pretreatment did not inhibit degradation of IkappaB-alpha or -beta in response to stimulation, or prevent the increase in NF-kappaB binding activity shown by EMSA. However, dexamethasone significantly inhibited the increase in monocyte chemoattractant protein-1 mRNA seen after stimulation with interleukin 1beta, although this was not complete. It did not reduce transcription of an NF kappaB reporter. In comparison, the pyrrolidine derivative of dithiocarnamate (PDTC), a known inhibitor of NF-kappaB, prevented the increase in NF-kappaB binding activity and significantly reduced transcription of the NF-kappaB reporter. These studies suggest that steroids can partially inhibit transcriptional activation by NF-kappaB in mesangial cells but not through an increase in IkappaB-alpha protein alone. Their effect must occur at the promoter and may be restricted to some NF-kappaB-responsive genes. Therapies that block NF kappaB more effectively than steroids in mesangial cells, therefore, may be useful in the treatment of mesangial proliferative glomerulonephritis. PMID- 9727371 TI - Differential regulation of matrix metalloproteinases and their inhibitors in human glomerular epithelial cells in vitro. AB - The present study examines the effect of transforming growth factor-beta1 (TGF beta1) and interleukin-1beta (IL-1beta) on the regulation of gelatinase A, gelatinase B, tissue inhibitor of metalloproteinase-I (TIMP-I) and TIMP-II in human glomerular epithelial cells (GEC). The addition of TGF-beta1 resulted in the increased production and secretion of both gelatinase A (72-kD type IV collagenase) and gelatinase B (92-kD type IV collagenase), in a dose- and time dependent manner. In contrast, the addition of IL-1beta to GEC resulted in the stimulation of secretion of gelatinase B but not gelatinase A. When the secretion of the regulatory inhibitors was examined, IL-1beta or TGF-beta1 both resulted in an increased secretion of TIMP-I, whereas the secretion of TIMP-II was downregulated. Such results demonstrate an independent and opposite regulation of the enzymes and their inhibitors. Of particular interest was the observation of the differential regulation of gelatinase A and its specific inhibitor TIMP-II (which binds to the latent form of this enzyme) in response to TGF-beta1. These results for the first time indicate that in human GEC, matrix metalloproteinases (MMP), as well as their specific inhibitors, are independently regulated by different cytokines. MMP and their regulatory tissue inhibitors (TIMP) play an important role in tissue remodeling. The results of the present study serve to emphasize both the complex regulation of matrix metabolism in the glomerulus and the potential pathologic role of an imbalance between the proteinases and their inhibitors in various forms of glomerular disease. PMID- 9727373 TI - Body size and risk of kidney stones. AB - A variety of factors influence the formation of calcium oxalate kidney stones, including gender, diet, and urinary excretion of calcium, oxalate, and uric acid. Several of these factors may be related to body size. Because men on average have a larger body size and a threefold higher lifetime risk of stone formation than women, body size may be an important risk factor for calcium oxalate stone formation. The association between body size (height, weight, and body mass index) and the risk of kidney stone formation was studied in two large cohorts: the Nurses' Health Study (NHS; n = 89,376 women) and the Health Professionals Follow-up Study (HPFS; n = 51,529 men). Information on body size, kidney stone formation, and other exposures of interest was obtained by mailed questionnaires. A total of 1078 incident cases of kidney stones in NHS during 14 yr of follow-up and a total of 956 cases in HPFS during 8 yr of follow-up were confirmed. In both cohorts, the prevalence of a stone disease history and the incidence of stone disease were directly associated with weight and body mass index. However, the magnitude of the associations was consistently greater among women. Specifically, the age-adjusted prevalence odds ratio for women with body mass index > or = 32 kg/m2 compared with 21 to 22.9 kg/m2 was 1.76 (95% confidence interval, 1.50 to 2.07), but 1.38 (95% confidence interval, 1.16 to 1.65) for the same comparison in men. For incident stone formation, the multivariate relative risks for the similar comparisons were 1.89 (1.51 to 2.36) for women and 1.19 (0.83 to 1.70) in men. Height was inversely associated with the prevalence of stone disease but was not associated with incident stone formation. These results suggest that body size is associated with the risk of stone formation and that the magnitude of risk varies by gender. Additional studies are necessary to determine whether a reduction in body weight decreases the risk of stone formation, particularly in women. PMID- 9727372 TI - All four putative ligand-binding domains in megalin contain pathogenic epitopes capable of inducing passive Heymann nephritis. AB - Megalin (gp330) is the main target antigen involved in the induction of Heymann nephritis (HN), a rat model of human membranous nephropathy. Its large extracellular region contains four putative ligand-binding domains separated by spacer regions. Previously, it was reported that the second ligand-binding domain (LBD II) of megalin is involved in the pathogenesis of passive HN because it is capable of binding antibodies in vivo and initiating formation of immune deposits (ID). This study explores the possibility that pathogenic epitopes might also be present in the other putative ligand-binding domains. Recombinant fragments of ligand-binding domains (LBD) I through IV expressed in a baculovirus system were used to generate polyclonal domain-specific antibodies. Antibodies raised against each of the recombinant megalin fragments reacted preferentially with its respective antigen and with whole megalin by immunoblotting. Each of the antibodies also gave a characteristic brush-border staining for megalin by indirect immunofluorescence on rat kidney. When rats were injected with the domain-specific antibodies to test their ability to produce passive HN, glomerular ID were present in kidneys of all injected animals. The staining pattern in glomeruli of rats injected with LBD I, III, or IV was similar to that obtained with antibodies to LBD II. It is concluded that passive HN can be induced with antibodies against LBD I, III, and IV, as well as LBD II, and that each of the ligand-binding domains contains a pathogenic epitope. These findings provide further evidence for the multiple epitope model of HN. PMID- 9727374 TI - Association between the angiotensin-converting enzyme-insertion/deletion polymorphism and diabetic nephropathy: a methodologic appraisal and systematic review. AB - Recent studies have implicated a variant of the angiotensin-converting enzyme gene (ACE), associated with increased activity of this enzyme, in the development and progression of diabetic nephropathy. This study provides a systematic review of all cross-sectional, case-control, and cohort studies in patients with insulin dependent (IDDM) or non-insulin-dependent (NIDDM) diabetes mellitus of any race, examining the relationship between the ACE-insertion/deletion polymorphism and nephropathy. Nineteen studies in 21 populations published between 1994 and 1997 presenting data on 5336 patients were reviewed. Two investigators independently assessed the studies on methodologic quality, performance of study, and association between the ACE-insertion/deletion polymorphism and nephropathy. Separate analyses of the relationship between genotype and allele frequencies were performed for patients with IDDM and NIDDM by race, using Peto's odds ratio. In Caucasians with IDDM, pooling was not performed due to heterogeneity of the studies, but among the homogeneous studies, no association was detected. Likewise, no association was observed in Caucasian patients with NIDDM (odds ratio [OR], 1.10; 95% confidence interval [95% CI], 0.83 to 1.45). In Asian patients with NIDDM, the risk of nephropathy was increased in the presence of the DD or ID genotype (OR, 1.88; 95% CI, 1.42 to 2.85). Although this analysis fails to confirm an association between the ACE-insertion/deletion genotype and nephropathy in Caucasians with NIDDM or IDDM, a role for this genetic marker in Asian patients cannot be ruled out. However, due to methodologic limitations of individual studies, no definite conclusions can be drawn from this analysis. Clearly, more rigorous methodology needs to be applied in future studies. PMID- 9727375 TI - Angiotensin I-converting enzyme gene polymorphisms: relationship to nephropathy in patients with non-insulin dependent diabetes mellitus. AB - Nephropathy is a frequent complication of long-term diabetes. Strong evidence exists that genetic predisposition plays a major role in the development of diabetic nephropathy. The role of the angiotensin I-converting enzyme gene (ACE) in the susceptibility to nephropathy in diabetes, especially in non-insulin dependent diabetes mellitus (NIDDM), remains unclear. This study examines the association of two ACE polymorphisms: a 287-bp insertion/deletion (I/D) in intron 16 and PstI (A/G substitution in intron 7; alleles P/M) with renal complications in 941 NIDDM patients. From this group, for further analysis 127 patients were selected with overt proteinuria or chronic renal failure, 335 patients with microalbuminuria, and a control group of 254 normoalbuminuric patients with a diabetes duration of at least 10 yr. No significant differences in the distribution of ACE I/D and PstI genotypes or allele frequencies were observed between the examined groups. The results of this study strongly suggest that there is no association between the ACE gene I/D and PstI polymorphisms and nephropathy in NIDDM. PMID- 9727376 TI - Ceramide accumulation during oxidant renal tubular injury: mechanisms and potential consequences. AB - Ceramide is an important signaling molecule that is typically generated via sphingomyelinase (SMase)-mediated sphingomyelin (SM) hydrolysis. Although diverse forms of renal injury elicit ceramide accumulation, the molecular determinants of this change and its contribution to tissue damage are poorly defined. The present study uses iron (Fe/hydroxyquinoline)-mediated injury of cultured human proximal tubular (HK-2) cells to gain additional insights into these issues. A 4-h Fe exposure doubled ceramide levels in the absence of cell death. This was independent of de novo synthesis, since ceramide synthase inhibition (with fumonisin B1) had no effect. Oxidant stress directly suppressed, rather than stimulated, SMase activity by: (1) decreasing SMase levels; (2) depleting SMase stimulating glutathione; and (3) increasing SM resistance to SMase attack. Fe suppressed cell sphingosine levels (3 to 4 times ceramide/sphingosine ratio increments), suggesting a possible ceramidase block. Fe did not directly affect HK-2 ceramidase levels. However, arachidonic acid (C20:4) accumulation, a consequence of oxidant-induced phospholipase A2 (PLA2) activation, markedly suppressed ceramidase and stimulated SMase activity. Exogenous C20:4, as well as PLA2 (in doses simulating Fe-induced deacylation) recapitulated Fe's ceramide generating effect. Because C20:4 is directly cytotoxic, it was hypothesized that ceramide might offset some of C20:4's adverse effects. Supporting this possibility were the following: (1) C20:4 exacerbated Fe toxicity; (2) this was abrogated by ceramide treatment; and (3) ceramide blunted Fe-mediated cell death. CONCLUSIONS: (1) ceramide accumulation during acute cell injury can be an adaptive response to PLA2 activation/C20:4 generation; (2) C20:4-induced ceramidase inhibition, coupled with SMase stimulation, may trigger this result; and (3) these ceramide increments may exert a "biostat" function, helping to offset C20:4/PLA2- and "catalytic" iron-mediated tubular cell death. PMID- 9727377 TI - Plasma levels of pentosidine in diabetic patients: an advanced glycation end product. AB - Nonenzymatic reactions between glucose and proteins yield advanced glycation end products (AGE) such as pentosidine. AGE accumulate in diabetic patients, alter the structure and function of tissue proteins, stimulate cellular response, and have thus been implicated in diabetic tissue damage. The present study was undertaken to assess the factors determining plasma total pentosidine level in diabetic patients and the possible relation between plasma pentosidine level and diabetic complications. In diabetic patients, including patients with renal failure, plasma pentosidine levels, assessed by HPLC assay, were correlated with serum creatinine (P < 0.0001). In patients with normal renal function, pentosidine levels were correlated with blood glucose control (hemoglobin Alc: P = 0.0028; fructoselysine: P = 0.0133), serum creatinine (P = 0.029), patient age (P = 0.0416), duration of diabetes (P = 0.0431), and total cholesterol (P = 0.0056) and LDL-cholesterol (P = 0.0208). Multiple regression analysis revealed an independent influence of hemoglobin Alc and serum creatinine on pentosidine levels (r2 = 0.216, P = 0.0026). Pentosidine levels were higher in patients with than in those without hypertension (P = 0.043) or ischemic heart diseases (P = 0.0061). No such differences were observed between patients with and without albuminuria or retinopathy. Multiple regression analysis revealed an independent influence of plasma pentosidine on the presence of hypertension (r2 = 0.129, P = 0.0382) and of plasma pentosidine and HDL-cholesterol on the presence of ischemic heart disease (r2 = 0.326, P = 0.0012). The present study demonstrated that plasma pentosidine level was significantly influenced by the quality of glycemic control and renal function. Pentosidine level was also correlated with hypertension and ischemic heart disease, and might be taken as a biomarker of diabetic cardiovascular risk. PMID- 9727378 TI - Production of proinflammatory and regulatory monokines in hemodialysis patients shown at a single-cell level. AB - Immunologic complications of chronic renal failure are associated with the overproduction of proinflammatory cytokines by monocytes. This is partly due to renal failure itself but is further enhanced by hemodialysis treatment with frequent contact between blood and dialyzer membranes. Previous studies have shown an imbalance of proinflammatory and regulatory monokines in these patients. This study examines monokine production in hemodialysis patients using for the first time a very sensitive method of cytokine detection at a single-cell level by flow cytometry ("cytoflow technique"). Monocytes were stained intracellularly for the production of interleukin-6 (IL-6) and IL-10 after 20 h of culture with lipopolysaccharide. It was shown that high levels of proinflammatory IL-6 in hemodialysis patients are due to an increased number of monocytes producing this cytokine, while IL-6 synthesis per cell remains unchanged. In contrast, elevated levels of regulatory IL-10 are due to an increased synthesis per cell. This study demonstrates that in healthy subjects there is a population of monocytes producing exclusively IL-10 after 20 h of stimulation by lipopolysaccharide. This distinct population of regulatory monocytes is infrequent in dialysis patients, in whom most of the IL-10-positive monocytes also produce IL-6. These findings indicate that overproduction of proinflammatory factors in dialysis patients is at least in part due to a loss of cytokine-specific differentiation in monocytes. PMID- 9727379 TI - Clinical practice guidelines: prevention of cytomegalovirus disease after renal transplantation. AB - OBJECTIVE: To develop a set of comprehensive, standardized, evidence-based guidelines for the use of antiviral therapy to prevent cytomegalovirus disease in adult patients having undergone renal transplantation. OPTIONS: The use of medication, at the time of induction therapy or at the earliest sign of viremia. Treatments were evaluated by patient and donor serologic groups and the induction regimen used. OUTCOMES: The control of symptoms and features of cytomegalovirus disease over the first 6 mo to 1 yr after transplantation. EVIDENCE: Articles, compiled using a MEDLINE search from 1976 to July 1997, were reviewed by representatives of nephrology, microbiology, pharmacy, and epidemiology. Additional information was obtained from recent review articles and conference abstracts, and from experts in the field. VALUES: The evidence-based methods and values of the Canadian Task Force on the Periodic Health Examinations were used. High value was placed on studies with a randomized controlled design and blinded outcome observers. Study quality was classified as poor when cointervention was present (especially with regard to immunosuppressive regimens), when more than 20% of patients were lost to follow-up, and when intention to treat analysis was not performed. Recommendations were made with a graded system (grades A and B: Use of the intervention advised, based on high or fair quality evidence, respectively; grades D and E: Use of the intervention not advised, based on high or fair quality evidence, respectively: grade C: No recommendation made because of insufficient or conflicting evidence). RECOMMENDATIONS: (1) Seropositive recipient; donor seropositive or seronegative; immunosuppression with antilymphocyte products. Prophylaxis with antiviral therapy recommended (grade A recommendation). (2) Seronegative recipient; seropositive donor; immunosuppression with antilymphocyte products. Prophylaxis with antiviral therapy recommended (grade A recommendation) (3) Seronegative recipient; seropositive donor; conventional immunosuppression. Prophylaxis with antiviral therapy recommended (grade B recommendation). (4) Seronegative recipient; seronegative donor; any immunosuppressive regimen. No prophylaxis with antiviral therapy required (grade D/E recommendation). (5) Seropositive recipient: donor seropositive or seronegative; conventional immunosuppression. Prophylaxis left to the discrimination of the physician in charge (grade C recommendation). PMID- 9727380 TI - Antihuman immunoglobulin affinity immunoadsorption strongly decreases proteinuria in patients with relapsing nephrotic syndrome. AB - Approximately 20 to 30% of patients with idiopathic nephrotic syndrome and focal glomerulosclerosis experience a relapse of their nephrotic syndrome after transplantation. Previously, it has been shown that ex vivo immunoadsorption on protein A strongly (although transiently) reduces proteinuria in relapsing patients. To investigate whether the factor(s) that give rise to albuminuria are bound directly to protein A in the immunoadsorption procedure or are part of a complex with Ig, four patients with relapse of focal glomerulosclerosis presenting as nephrotic syndrome after transplantation were treated, sequentially, using a (non-protein A) anti-Ig affinity column and a protein A column. This study reports that the effect on proteinuria of immunoadsorption using an anti-Ig immunoaffinity column is comparable in its magnitude and kinetics to that of immunoadsorption on protein A. The two procedures were also equally effective in depleting the relapsing patients' plasma of a factor capable of altering the albumin permselectivity of isolated glomeruli in vitro. This study demonstrates for the first time that immunoglobulins have a role in the nephrotic syndrome. In addition, the fact that the two different immunoadsorption procedures both resulted in the removal of the same putative albuminuric factor in these patients and that no autoreactivity of eluted immunoglobulins was observed on human tissues strongly suggests that the factor or factors that may be responsible for immediate nephrotic syndrome after transplantation are bound to an immunoglobulin. However, no firm evidence can be yet provided against a direct role of immunoglobulins. PMID- 9727381 TI - Novel angiotensin peptides regulate blood pressure, endothelial function, and natriuresis. AB - Accumulating evidence suggests that angiotensin-(1-7) is an important component of the renin-angiotensin system, having actions that are either identical to or opposite that of angiotensin II. Angiotensin I can be directly converted to angiotensin-(1-7), bypassing formation of angiotensin II. This pathway is under the control of three enzymes: neutral endopeptidases 24.11 (neprilysin) and 24.15 and prolyl-endopeptidase 24.26. Two of the three angiotensin-forming enzymes (neprilysin and endopeptidase 24.15) also contribute to the breakdown of bradykinin and the atrial natriuretic peptide. Furthermore, angiotensin-(1-7) is a major substrate for angiotensin-converting enzyme. These observations suggest that the process of biotransformation between the various Ang peptides of the renin-angiotensin system and other vasodepressor peptides are intertwined through this enzymatic pathway. Substantial evidence suggests that angiotensin-(1-7) stimulates the synthesis and release of vasodilator prostaglandins, and nitric oxide, while also augmenting the metabolic actions of bradykinin. In addition, angiotensin-(1-7) alters tubular sodium and bicarbonate reabsorption, decreases Na+-K+-ATPase activity, induces diuresis, and exerts a vasodilator effect. These physiologic effects of angiotensin-(1-7) favor a blood pressure-lowering effect. The majority of the data currently available suggest that angiotensin-(1-7) mediates its effects through a novel non-AT1/AT2 receptor subtype. PMID- 9727382 TI - Beta-2 microglobulin in renal disease. PMID- 9727383 TI - Alport syndrome and thin glomerular basement membrane disease. PMID- 9727384 TI - Cyclosporin A in patients receiving renal allografts from cadaver donors. 1978. PMID- 9727385 TI - A-53930A and B, novel N-type Ca2+ channel blockers. AB - A-53930A, B and C, which inhibit N-type Ca2+ channels, were isolated from the culture broth of Streptomyces vinaceusdrappus SANK 62394. A-53930A and B were new compounds which contained a carbamoyl group on the 6-hydroxyl group of the D gulosamine part of streptothricin. A-53930C was identical to streptothricin B. A 53930A, B and C inhibited [125I]omega-conotoxin MVIIA binding to N-type Ca2+ channels (IC50= 0.17, 0.091 and 0.071 microM), but did not inhibit [3H]PN200-110 binding to L-type Ca2+ channels (IC50 > 50 microM). These compounds also inhibited [3H]norepinephrine release from chick cerebral cortex synaptosomes (IC50=91.0, 20.6 and 39.5 microM), indicating these compounds selectively block N type Ca2+ channels which are important for neurotransmitter release. It was also revealed that although A-53930C had antimicrobial activity against gram-negative and -positive bacteria and fungi, A-53930A and B showed weak activity only against gram-negative bacteria. PMID- 9727387 TI - Erabulenols, inhibitors of cholesteryl ester transfer protein produced by Penicillium sp. FO-5637. II. Structure elucidation of erabulenols A and B. AB - Structures of erabulenols A and B, novel fungal inhibitors of cholesteryl ester transfer protein were elucidated by spectroscopic studies including various NMR measurements. Erabulenols consist of a phenalenone skeleton and a 1,2,2 trimethyltetrahydrofuran moiety in common. Erabulenol B possesses an additional 2,6-dihydroxy-5-methyl-3-methylketonyl benzyl moiety. The absolute stereochemistry at the C-2' position of erabulenol A was deduced as S by comparison of the optical rotation with that of other related compounds. PMID- 9727386 TI - Erabulenols, inhibitors of cholesteryl ester transfer protein produced by Penicillium sp. FO-5637. I.Production, isolation and biological properties. AB - Penicillium sp. FO-5637, a soil isolate, was found to produce a series of inhibitors of cholesteryl ester transfer protein (CETP). Novel active compounds, designated erabulenols A and B, were isolated from the fermentation broth of the producing strain by solvent extraction, ODS column chromatography and HPLC. Erabulenols A and B inhibit human CETP activity with IC50 values of 47.7 and 58.2 microM in an in vitro assay system containing 200 microM BSA, respectively. PMID- 9727388 TI - TMC-52A to D, novel cysteine proteinase inhibitors, produced by Gliocladium sp. AB - New cysteine proteinase inhibitors, TMC-52A, B, C, and D, were isolated from the fungal fermentation broth. On the basis of a taxonomical study, the producing strain, F-2665, was characterized as Gliocladium sp. Spectroscopic analyses and chemical degradation have shown TMC-52A to D to be epoxysuccinyl peptides. TMC 52A to D strongly inhibited cysteine proteinases, in particular, cathepsin L with IC50 values of 13 nM, 10nM, 10nM, and 6nM, respectively. PMID- 9727389 TI - Hongoquercins A and B, new sesquiterpenoid antibiotics: isolation, structure elucidation, and antibacterial activity. AB - Two new sesquiterpenoid antibiotics, hongoquercins A and B, were isolated from the extracts of an unidentified fungus. The structures of both metabolites were determined by spectroscopic analysis. They are related to a class of compounds commonly found in brown algae and dictyoceratid sponges. Hongoquercin A exhibited moderate activity against gram-positive bacteria. PMID- 9727390 TI - New types of liposidomycins that inhibit bacterial peptidoglycan synthesis and are produced by Streptomyces. I. Producing organism and medium components. AB - Liposidomycins are atypical lipid-bearing nucleoside antibiotics that inhibit bacterial peptidoglycan synthesis. A producing strain was identified as a Streptomyces sp. from its cultural characteristics and physiological properties. It produced new types of liposidomycins that lacked sulfate and/or 3 methylglutaric acid moieties present in known liposidomycins by changing medium components. Sucrose and malt extract were particularly suitable sources for specific production of the new types of liposidomycins. PMID- 9727391 TI - New types of liposidomycins that inhibit bacterial peptidoglycan synthesis and are produced by Streptomyces. II. Isolation and structure elucidation. AB - Various new liposidomycins were isolated from a culture of the strain Streptomyces sp. SN-1061M by changing medium components and they were classified into four types (I-IV) based on their structures. They were purified by butanol extraction, silica gel and LH-20 column chromatographies, and high performance liquid chromatography on ODS columns. Type (I) has the original structure which has sulfate and 3-methylglutaric acid moieties. Type (II) has no 3-methylglutaric acid moiety and type (III) has no sulfate moiety. Type (IV) has neither moiety. Type (III) and (IV) compounds, which have no sulfate moiety, exhibited more potent antimicrobial activity. PMID- 9727392 TI - WA8242A1, A2 and B, novel secretary phospholipase A2 inhibitors produced by Streptomyces violaceusniger. III. Structure elucidation and total synthesis of WA8242B. AB - The structure of WA8242B, a potent novel inhibitor against phospholipase A2, was fully characterized by spectroscopic methods and chemical degradation. The success of total synthesis of WA8242B confirmed the structure and allowed the pharmacological study of WA8242B. The structures of WA8242A1 and A2 were also described. PMID- 9727393 TI - Characterization and optimization of in vitro assay conditions for (1,3)beta glucan synthase activity from Aspergillus fumigatus and Candida albicans for enzyme inhibition screening. AB - (1,3)Beta-D-glucan synthase (E.C.2.4.1.34. UDP-glucose: 1,3-beta-D-glucan 3-beta glucosyl transferase) catalyzes the polymerization of glucose ([1-3]-beta linkages) using UDP-glucose as substrate. We have determined optimal in vitro conditions for the assay of (1,3)beta-glucan synthase activity from Aspergillus fumigatus and Candida albicans. These included lysis of cells in the following for C. albicans, 100 mM HEPES, pH 8.0, 10 microM guanosine 5'-O-(3 thiotriphosphate) (GTPgammaS), 2 mM ethylenediaminetetraacetic acid (EDTA), disodium salt, 5 mM NaF, 250 mM sucrose, and 10 mM NaH2PO4; and for A. fumigatus, 50 mM HEPES, 10mM EDTA, 750 mM sucrose, 10 mM NaH2PO4, 100 mM cellobiose and 50 microM GTPgammaS. Resulting low-speed supernatants were used as enzyme sources to determine the optimal in vitro assay conditions. We have characterized the resulting enzyme activities and tested the optimized assays with known (1,3)beta glucan synthase inhibitors including cilofungin, papulacandin, aculeacin A, and echinocandin B. We have used both optimized assays to screen > 1000 extracts of marine macroorganisms and, using bioassay-guided purification, have identified (1,3)beta-glucan synthase inhibitors. PMID- 9727394 TI - Studies on the biosynthesis of terpenoid compounds produced by actinomycetes. 3. Biosynthesis of the isoprenoid side chain of novobiocin via the non-mevalonate pathway in Streptomyces niveus. PMID- 9727395 TI - 3'-Demethoxy-3'-hydroxystaurosporine-O-methyltransferase from Streptomyces longisporoflavus catalyzing the last step in the biosynthesis of staurosporine. PMID- 9727396 TI - FK041, a novel orally active cephalosporin: synthesis and biological properties. PMID- 9727397 TI - Enantioselective and convergent synthesis of the 20-membered lactam aglycon of vicenistatin antitumor antibiotic. PMID- 9727398 TI - Nonvascular contractile cells in sclera and choroid of humans and monkeys. AB - PURPOSE: To investigate by histochemistry and immunohistochemistry the distribution and innervation of nonvascular contractile cells in the sclera and choroid of humans and monkeys. METHODS: Globes were obtained from 2 macaque monkeys and 19 human cadavers that ranged in age from fetal life to 94 years. Immunohistochemistry was performed using monoclonal antibody against human smooth muscle (SM) alpha-actin and tyrosine hydroxylase (TH). The nicotinamide-adenine dinucleotide phosphate (NADPH)- diaphorase reaction was used as a marker for nitric oxide synthase. RESULTS: The scleras of all but fetal, newborn, and infant globes exhibited myofibroblasts, amelanotic, fibroblastlike cells having SM alpha actin immunoreactivity. In the choroid of all but fetal eyes, SM cells were present in the suprachoroidal layer, forming a reticulum of flattened laminae, and in the choriocapillaris where ovoid-to-spindle-shaped SM cells were arrayed in parallel layers immediately adjacent to Bruch's membrane. Contractile cells in the sclera and choroid were most concentrated subfoveally and were sparse anteriorly. Nerve terminals positive for NADPH- diaphorase were colocalized with SM alpha-actin-positive cells in the sclera and choroid, whereas TH-positive nerve terminals colocalized with SM cells in the choroid. Clusters of ganglion cells were present on the posterior surface of globes near SM cells. CONCLUSIONS: The posterior choroid and sclera of humans and monkeys contain nonvascular contractile cells. The presence of nerve terminals and adjacent ganglion cells suggests neural control of these contractile cells. The absence of such contractile cells in fetal, newborn, and infant eyes is an argument against a major role of these cells in promoting ocular enlargement. These contractile cells may instead participate in regulation of refractive state by maintenance of ocular size in the face of intraocular pressure or in intermediate-term regulation of choroidal thickness. PMID- 9727399 TI - Canine cone transducin-gamma gene and cone degeneration in the cd dog. AB - PURPOSE: To characterize the cDNA and the organization of the gene encoding the cone-specific gamma subunit of transducin (Tgamma c) and to examine this gene as a candidate for the recessively inherited cone photoreceptor degeneration in the cd dog. METHODS: Canine Tgamma c cDNA was cloned and sequenced. Polymerase chain reaction (PCR) was used to define the Tgamma c gene structure, northern blot analysis to examine the level of expression of Tgamma c mRNA in control and cd affected retinas, and immunocytochemistry to determine the presence and localization of Tgamma c in normal and cd retinas. RESULTS: Immunocytochemical results showed Tgamma c localized to cone photoreceptor outer segments in the normal retina, whereas no Tgamma c immunoreactivity was observed in the cd retinas. However, the level of transcription and the primary structure of the cloned cDNA coding for the 69-amino acid protein were identical in retinas from wild-type and affected dogs. CONCLUSIONS: Although Tgamma c immunoreactivity was specifically absent in the cd dog retina, no differences were detected between normal and cd retinas in the nucleotide sequence of Tgamma c mRNA or in its synthesis. These results indicate that a mutation in the Tgamma c gene may not be causally associated with the cd dog disease. These findings suggest that possible abnormalities in posttranslational modification of Tgamma c or defective assembly of the transducin alphabetagamma complex could lead to rapid degradation of Tgamma c. PMID- 9727400 TI - Molecular cloning of a new angiopoietinlike factor from the human cornea. AB - PURPOSE: To isolate tissue-specific gene products that contribute to corneal integrity. METHODS: A cDNA library was constructed and differentially hybridized. Cornea-specific clones were purified and further characterized. RESULTS: In this study cornea-specific gene products were isolated by differential cDNA hybridization. In addition to known cornea-specific gene products, a transcript was isolated coding for a protein homologous to the angiopoietins, a recently described family of (anti)angiogenic factors. Subsequently, the full cDNA was sequenced, and the identified open reading frame was named cornea-derived transcript 6 (CDT6). Similar to the angiopoietins, CDT6 contains a hydrophobic NH2-terminal sequence, a coiled-coil domain, and a COOH-terminal fibrinogenlike domain. Expression of CDT6 could be detected only in the cornea and not in several other adult human tissues. Within the cornea, expression of CDT6 is confined to the stromal layer. CONCLUSIONS: The human cornea shows high-level expression of a gene product homologous to the (anti)angiogenic factors, the angiopoietins. This homology, together with stromal-specific expression, suggests that this factor may contribute to the avascularity of the human cornea. PMID- 9727401 TI - Isolation and characterization of human cathepsin V: a major proteinase in corneal epithelium. AB - PURPOSE: To isolate and characterize a novel cathepsin gene, as part of the systematic isolation of genes uniquely active in corneal epithelium. METHODS: For the isolation of a full-length cDNA clone, a probe was selected from a set of expressed sequence tag clones classified as unique to corneal epithelium. Inserted cDNA was introduced into insect cells using a baculovirus expression system, and the secretion of recombinant protein was identified using antisera against a synthetic peptide. Proteolytic activity was determined using bovine serum albumin (BSA) as substrate. The expressions of the novel cathepsin in human cornea and other tissues were examined by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The full-length cDNA clone encoded a peptide of 334 amino acids with 82% identity with bovine cathepsin L and 77% identity with human cathepsin L when aligned. The recombinant protein produced in the baculovirus expression system cleaves BSA, and its activity was inhibited by the cysteine proteinase inhibitors E-64 and leupeptin, but not by pepstatin A, phenylmethylsulfonyl fluoride, and EDTA. By RT-PCR, a low level of expression was observed in some other epithelial tissues of ectodermal origin, but only in cornea was it higher than cathepsin L, which is known to be a general lysosomal cathepsin. Cathepsin V protein was detected in human corneal epithelium by western blot analysis, but not in tear fluid. CONCLUSIONS: The amino acid homology and proteolytic activity of the recombinant protein indicate that the novel gene is a new member of the cathepsins that have features of cysteine proteinase. Its uniquely high expression in corneal epithelium strongly implies an important role in corneal physiology. PMID- 9727402 TI - Myopes show increased susceptibility to nearwork aftereffects. AB - PURPOSE: Some aspects of accommodation may be slightly abnormal (or different) in myopes, compared with accommodation in emmetropes and hyperopes. For example, the initial magnitude of accommodative adaptation in the dark after nearwork is greatest in myopes. However, the critical test is to assess this initial accommodative aftereffect and its subsequent decay in the light under more natural viewing conditions with blur-related visual feedback present, if a possible link between this phenomenon and clinical myopia is to be considered. METHODS: Subjects consisted of adult late- (n = 11) and early-onset (n = 13) myopes, emmetropes (n = 11), and hyperopes (n = 9). The distance-refractive state was assessed objectively using an autorefractor immediately before and after a 10 minute binocular near task at 20 cm (5 diopters [D]). RESULTS: Group results showed that myopes were most susceptible to the nearwork aftereffect. It averaged 0.35 D in initial magnitude, with considerably faster posttask decay to baseline in the early-onset (35 seconds) versus late-onset (63 seconds) myopes. There was no myopic aftereffect in the remaining two refractive groups. CONCLUSIONS: The myopes showed particularly striking accommodatively related nearwork aftereffect susceptibility. As has been speculated and found by many others, transient pseudomyopia may cause or be a precursor to permanent myopia or myopic progression. Time-integrated increased retinal defocus causing axial elongation is proposed as a possible mechanism. PMID- 9727403 TI - Isolation, culture, and characterization of endothelial cells from Schlemm's canal. AB - PURPOSE: An important goal in glaucoma research has been to understand the functional contribution of trabecular meshwork (TM) and Schlemm's canal (SC) endothelia to aqueous humor outflow resistance. To date, TM cells are routinely cultured and used as a model by several laboratories. However, there has been only limited success in isolating SC cells. The current objective was to develop a technique for selective isolation and culture of endothelial cells from human SC. METHODS: The anterior chamber of human cadaveric eyes was cut into eight equal and radially symmetric wedge-shaped pieces. Using a dissecting microscope, a gelatin-coated suture (6-0 sterile nylon monofilament) was gently inserted into the lumen of SC and advanced into the canal. The cannulated pieces of tissue were placed in culture medium and maintained for 3 weeks. Sutures were removed from SC and cells seeded onto 3-cm culture plates. Morphology, growth characteristics, and expression of endothelial surface antigens and other cellular markers were evaluated. RESULTS: Of the 20 pairs of eyes that were cannulated, primary cells were obtained from 13. All SC cell isolates had a fusiform morphology; formed nonoverlapping, linearly arranged monolayers; and were contact inhibited. Schlemm's canal cell isolates reacted with antibodies specific for CD44 (hyaluron receptor), CD54 (intercellular adhesion molecule-1, ICAM-1), tissue-type plasminogen activator, and TM-inducible glucocorticoid-responsive protein myocilin (TIGR-MYOC). Unlike TM cells, however, TIGR-MYOC protein was not induced in SC cells after long-term dexamethasone treatment. Schlemm's canal cells endocytosed low-density lipoprotein and acetylated low-density lipoprotein, and in the presence of Matrigel organized into multicellular tubelike structures. CONCLUSIONS: Cannulation of SC with gelatin-coated suture material is an effective method for the isolation of human SC cells and provides a cellular model to study the potential role of SC cells in aqueous humor outflow function. PMID- 9727404 TI - Effect of beta-adrenergic agonists on paracellular width and fluid flow across outflow pathway cells. AB - PURPOSE: To determine whether the adrenergic agonists epinephrine and isoproterenol regulate fluid flow across endothelial cells cultured from the human aqueous outflow pathway and to evaluate associated cellular mechanisms. METHODS: Confluent monolayers of human trabecular meshwork (TM) or Schlemm's canal endothelial (SCE) cells were grown on porous filter supports. The monolayers were perfused with media while fluid flow, expressed as hydraulic conductivity (HC = microl/min/mm Hg/cm2), was continuously measured in preparations treated with isoproterenol, epinephrine, or control medium. Morphometric ultrastructural methods were used to measure the area occupied by the intercellular space and by each cell. RESULTS: SCE cells and TM cells exposed to isoproterenol or epinephrine responded with an increase in transendothelial fluid flow. Dose-response curves for both adrenergic agonists showed that HC increased linearly as a function of the log of the isoproterenol and epinephrine concentration. At 10(-4) M isoproterenol, the HC increased threefold, and threshold conditions were reached at 10(-9) M. The increase in HC was apparent after isoproterenol had been applied for 1 hour, reached a peak in 3 to 4 hours, and declined gradually to return to baseline conditions in 10 to 12 hours. Morphometric analyses showed that the area occupied by the intercellular space increased fourfold when isoproterenol was used at 10(-4) M, whereas the cell area decreased as a function of the concentration of adrenergic agonist. Epinephrine's effects on HC and cell morphology were blocked by pretreatment with equimolar concentrations of the nonselective beta-blocker, timolol. CONCLUSIONS: Epinephrine and isoproterenol increase flow through the paracellular pathway of SCE and TM cells through a beta-receptor mediated response that widens the intercellular space and reduces cell area. These findings support the hypothesis that epinephrine decreases the intraocular pressure in glaucoma therapy by promoting fluid flow across the SCE and TM cells lining tissues of the major aqueous outflow pathway. PMID- 9727405 TI - Analysis of immune deviation elicited by antigens injected into the subretinal space. AB - PURPOSE: To determine whether the subretinal space supports the induction of deviant immune responses to cell-associated and soluble antigens and to elucidate factors influencing the immunologic properties of the subretinal space. METHODS: P815 mastocytoma cells were used as cell-associated antigens and were inoculated into the anterior chamber (AC), the vitreous cavity (VC), the subretinal space, and subconjunctivally in H2-compatible, but minor H-incompatible, BALB/c mice. Ovalbumin, as a soluble antigen, was similarly injected into eyes, after which recipient animals were immunized with ovalbumin and complete Freund's adjuvant. Delayed-type hypersensitivity (DTH) was assessed by ear challenge. To alter the conditions in the subretinal space, the outer blood-retinal barrier was disrupted by compromising retinal pigment epithelial (RPE) cells with a systemic injection of sodium iodate. Immune privilege in the AC was abolished by mild corneal cauterization. RESULTS: Antigen-specific DTH did not develop in mice in which alloantigenic tumor cells or ovalbumin was injected into the AC, the VC, or the subretinal space, and the mice's spleens contained lymphocytes capable of suppressing DTH when adoptively transferred into naive mice. When RPE cells were compromised with sodium iodate, tumor cells or ovalbumin injected into the subretinal space or the VC did not induce immune deviation, although the AC of these eyes still promoted AC-associated immune deviation. By contrast, when immune privilege in the AC was abolished by corneal cauterization, neither tumor cells nor ovalbumin injected into the subretinal space or the VC of eyes elicited immune deviation. CONCLUSIONS: The subretinal space supports immune deviation for histoincompatible tumor cells and soluble protein antigens by actively suppressing antigen-specific DTH. Acute loss of immune privilege in the subretinal space and the VC does not cause loss of privilege in the AC, but abolition of immune privilege in the AC eliminates the capacity of the subretinal space and the VC to support immune deviation to antigens injected locally. PMID- 9727406 TI - Conditions affecting enhanced corneal allograft survival by oral immunization. AB - PURPOSE: To determine the optimal conditions for enhancing corneal allograft survival by oral immunization with donor-specific alloantigens. METHODS: CB6F1 mice were orally immunized with various doses of C3H/Hej corneal epithelial and endothelial cells before receiving orthotopic C3H/Hej corneal allografts. Paraformaldehyde-fixed corneal cells were compared with viable corneal cells for their capacity to promote corneal allograft survival. The mucosal adjuvant, cholera toxin B (CTB), was examined for its capacity to enhance corneal graft survival when given separately or conjugated to corneal cells used for oral immunization. Oral immunization was also evaluated for its capacity to prevent immunologic rejection in three high-risk settings: preimmunized hosts, hosts with prevascularized graft beds, and grafts that contain donor-specific Langerhans' cells. RESULTS: Optimal graft survival occurred when 2 x 10(6) corneal cells were administered orally 10 days before orthotopic corneal transplantation. Paraformaldehyde-fixed corneal cells were as effective as viable cells in preventing corneal graft rejection. Cholera toxin B enhanced the efficacy of oral immunization when conjugated with the orally administered corneal cells but was ineffectual when administered separately. Oral immunization with donor corneal cells enhanced corneal graft survival in all three high-risk settings. CONCLUSIONS: Oral immunization with donor cells is an effective strategy for enhancing corneal graft survival and preventing graft rejection in high-risk settings. Graft enhancement is optimized when the orally administered cells are conjugated with CTB and administered before corneal transplantation. Because fixed cells retain their capacity to enhance corneal graft survival, it may be possible to store donor cells for long-term use in high-risk hosts. PMID- 9727407 TI - Tissue-specific accumulation of latency-associated transcripts in herpes virus infected rabbits. AB - PURPOSE: Herpes simplex virus (HSV) DNA persists in the corneas of patients and animals with a history of herpetic keratitis. The purpose of this study was to detect viral transcripts in the corneas of latently infected rabbits with a history of herpetic keratitis to determine whether the viral DNA represents latent virus, characterized by the restricted transcription of HSV genes and accumulation of the stable latency-associated transcripts (LATs), as occurs in neurons. METHODS: Rabbits were injected in the subalveolar mucosa with HSV strain RE. After 30 days, corneas were infected by intrastromal injection of HSV. Corneal disease was evaluated, and 7 to 378 days after infection, the rabbits were killed. DNA and RNA were isolated from corneas and trigeminal ganglia and amplified by PCR using gene-specific primers. RESULTS: Herpetic keratitis developed in all rabbits. All corneas of these immune rabbits contained viral DNA as many as 120 days after infection and then the frequency decreased over the next 260 days. Overall, viral DNA was detected in all ganglia and in 57% of corneas. All latently infected ganglia but no corneas contained LATs. Transcripts of the early viral gene for thymidine kinase were detected in 23 of 30 ganglia and 10 of 17 corneas. Transcripts for the late viral glycoprotein C were not detected in either tissue. CONCLUSIONS: These data document that after HSV keratitis, viral DNA persists in corneas in the absence of stable LATs and with restricted expression of other viral genes. PMID- 9727408 TI - Role of MIP-2 in neutrophil migration and tissue injury in the herpes simplex virus-1-infected cornea. AB - PURPOSE: Neutrophils are the most prominent cell type to migrate initially into the herpes simplex virus type 1 (HSV-1)-infected murine cornea. The role the C-X C chemokines macrophage inflammatory protein (MIP)-2 and KC play in promoting this response was investigated. METHODS: MIP-2 and KC were quantitated by enzyme linked immunosorbent assay. Neutralization of endogenous MIP-2 and KC was achieved by subconjunctival inoculation of the appropriate antibody. Infected corneas were examined immunohistochemically for infiltrating leukocytes and assayed for myeloperoxidase activity using the dye o-dianisidine. Depletion of neutrophils and natural killer cells was accomplished by intraperitoneal administration of RB6-8C5 and asialo GM1 antibodies. RESULTS: Herpes simplex virus type 1, when introduced intracorneally, stimulated the production of MIP-2 and KC, with peak synthesis occurring 48 hours after infection. Dose-response studies showed that exogenous MIP-2 was three to four times more potent than KC in attracting neutrophils as assessed by myeloperoxidase assay and immunohistochemical staining. Subconjunctival administration of neutralizing antibody to MIP-2 resulted in a sharp decrease in neutrophil infiltration and significantly reduced corneal opacity scores. In contrast, in vivo treatment with neutralizing antibody to KC did not suppress ocular inflammation. Additional studies indicated that MIP-2 and KC could be made by corneal epithelial cells and that production was promoted by interleukin (IL)-1. In vivo depletion of neutrophils sharply reduced MIP-2 levels but did not affect KC levels. CONCLUSIONS: Collectively, the results suggest that MIP-2 is the major chemokine that attracts neutrophils into the HSV-1 infected cornea, where the cells directly or indirectly cause tissue injury. Resident corneal cells and inflammatory cells contribute to MIP-2 synthesis, whereas KC production seems to be confined largely to corneal cells. PMID- 9727409 TI - Abnormal eye development associated with Cat4a, a dominant mouse cataract mutation on chromosome 8. AB - PURPOSE: Cat4a, one of four mutant alleles at the mouse Cat4 locus, causes central corneal opacity and anterior polar cataract in heterozygotes and microphthalmia in homozygotes. The Cat4 locus has been mapped to chromosome 8, 31 cM from the centromere. In this study ocular development of Cat4a mutant mice was investigated to characterize the defects in eye morphogenesis. METHODS: Serial sections from eyes of wild-type, heterozygous, and homozygous littermates were examined by means of light microscopy at selected intervals from embryonic day 11 to postnatal day 1. Eyes of adult heterozygous and homozygous mice also were evaluated histologically. RESULTS: Failure of separation of the lens vesicle from the surface ectoderm was the earliest structural defect observed. In heterozygous embryos, the abnormality was limited to persistent connection of the anterior pole of the lens to the cornea. Adult heterozygotes had defects in the central corneal stroma and endothelium and anterior polar cataracts with or without keratolenticular adhesion. In homozygous embryos, the persistent connection of lens to surface ectoderm was associated with aborted lens development, failure of closure of the optic fissure, and impairment of growth of the eyecup. Microphthalmic eyes of adult homozygous mice had a poorly developed cornea, and the anterior chamber and vitreous compartment were absent. An extensively folded retina and remnants of a degenerated lens filled the interior of the globe. CONCLUSIONS: A developmental defect inhibits separation of the lens vesicle from surface ectoderm in mice heterozygous or homozygous for the Cat4a mutation. In homozygotes subsequent lens and eye morphogenesis are also severely affected. Cat4a shows phenotypical similarity to several other independent mouse mutations including Small eye, a mutation of the Pax6 gene. Cat4 may be one of several genes involved in a common developmental path and may be part of the Pax6 regulated gene cascade governing eye morphogenesis. PMID- 9727410 TI - Neuropeptide Y and the retinal pigment epithelium: receptor subtypes, signaling, and bioelectrical responses. AB - PURPOSE: To characterize the potential for neuropeptide Y (NPY) signaling in the retinal pigment epithelium (RPE) by identifying the NPY receptor subtypes present, determining the effect of NPY on second-messenger production and membrane conductance, and establishing the neural retina as a site of NPY gene expression. METHODS: Neuropeptide Y receptors present in bovine and human RPE were identified using ribonuclease protection assays and reverse transcriptase coupled polymerase chain reaction. Assays of cyclic adenosine monophosphate (cAMP) and inositol phosphate production were performed using anion exchange chromatography and RPE cultures labeled with tritiated adenine or myo-inositol, respectively. Open-circuit recordings of transepithelial potential and resistance were performed using intact bovine RPE-choroid preparations. Neuropeptide Y expressing cells in the retina were identified by staining for beta-galactosidase activity in eyes from mice in which lacZ replaces a portion of the NPY gene. RESULTS: Human RPE contained transcripts encoding Y1, Y2, and Y5 receptors, the predominant subtypes present in the central nervous system. Bovine RPE contained transcripts encoding Y2 receptors but not Y1 receptors. However, cultured cells contained transcripts encoding Y1 and Y2 receptors. Neuropeptide Y signaling in cultured bovine RPE occurred predominately through the Y2 receptor subtype, because nanomolar amounts of NPY and NPY13-36, but not [Leu31,Pro34]NPY, significantly inhibited isoproterenol-induced cAMP accumulation. Apical application of NPY increased the transepithelial potential in RPE-choroid preparations. This response was greatly diminished after basolateral membrane Cl- channels were blocked or changes in intracellular Ca2+ concentration were prevented with a Ca2+ chelator. The NPY gene was expressed in amacrine cells of the inner nuclear and ganglion cell layers of the mouse retina. CONCLUSIONS: The discovery of functionally coupled NPY receptors in the RPE represents the identification of a novel site of expression of this receptor family. The effects of NPY on the electrophysiologic properties of the bovine RPE are consistent with a potential paracrine role in regulating basolateral membrane Ca2+-sensitive Cl- conductance linked to Cl- and fluid transport. PMID- 9727411 TI - In vivo cell tracking by scanning laser ophthalmoscopy: quantification of leukocyte kinetics. AB - PURPOSE: To image peripheral blood leukocyte traffic in the normal retinal and choroidal vasculature and to quantify the differences in the circulation dynamics between normal and concanavalin A (ConA)-activated leukocytes. METHODS: Normal or ConA-activated splenocytes were fluorescently labeled in vitro with 6 carboxyfluorescein diacetate (CFDA) and reinfused in vivo where they were tracked in the retinal and choroidal circulations of syngeneic rats by means of a scanning laser ophthalmoscope (SLO). Simultaneous digital and video images were captured for as long as 30 minutes, and the initial 15 seconds of image sequences and leukocyte dynamics were analyzed from digitized images by recording the velocity of trafficking cells and the number of stationary cells that accumulated with time, using a customized software package. RESULTS: Mean velocity (+/-SD) was 29.8 +/- 15.3 mm/sec in the retinal arteries, 14.7 +/- 7.2 mm/sec in the retinal veins, and 3.0 +/- 3.6 mm/sec in the retinal capillaries. Mean velocity in the choroidal vessels was 6.1 +/- 6.0 mm/sec. No significant difference in leukocyte velocity was found between activated and normal leukocytes in any of the vessel systems. However, activated leukocytes were observed to accumulate more within the choroidal vasculature (P < 0.001) and the retinal capillaries (P < 0.001) than in control animals, but not in larger retinal vessels. CONCLUSIONS: A technique to measure the kinetics of circulating leukocytes in vivo has been developed. Although leukocyte activation itself is insufficient to cause slowing of leukocyte velocity, the data indicate that leukocyte adherence to endothelium can be induced in the absence of local or systemic activating stimuli. PMID- 9727412 TI - Increases in retinovascular prostaglandin receptor functions by cyclooxygenase-1 and -2 inhibition. AB - PURPOSE: To determine the relative contribution of cyclooxygenase (COX)-1 and COX 2 in regulating prostaglandin (PG) E2 and PGF2alpha receptors (EP and FP, respectively) densities and their functions in retinal vasculature of neonatal pigs. METHODS: Newborn pigs were treated intravenously every 8 hours for 48 hours with saline, 40 mg/kg nonselective COX inhibitor ibuprofen, 80 mg/kg COX-1 inhibitor valeryl salicylate, or 5 mg/kg DuP697 and 5 mg/kg NS398, COX-2 inhibitors. Retinal microvessel EP and FP receptor densities were measured by radioligand binding and receptor-coupled effects by determining second-messenger inositol 1,4,5-trisphosphate (IP3) and vasomotor responses. Retinal blood flow (RBF) response to incremental increases in blood pressure (BP) was measured by a microsphere technique. RESULTS: Valeryl salicylate, DuP697, and NS398 reduced retinal PGE2 and PGF2alpha concentrations in the newborn by approximately half, whereas ibuprofen caused further reduction to levels observed in adults. Retinal vessel EP1, EP3, and FP receptor densities increased approximately threefold after treatments with COX-1 or COX-2 inhibitors, and five- to sixfold after ibuprofen treatment. EP and FP receptor upregulation was associated with corresponding increases in IP3 production and retinal vasoconstriction in response to PGF2alpha, fenprostalene (an FP agonist), PGE2, 17-phenyl trinor PGE2 (an EP1 agonist), and M&B28,767 (an EP3 agonist) and with enhanced RBF autoregulation of high BP (> or =125 mm Hg). Conversely, EP2 receptor density and coupled functions were minimally affected by COX inhibition. CONCLUSIONS: Data suggest that increased COX-1- and COX-2-catalyzed prostaglandin synthesis contribute equivalently to the downregulation of retinovascular EP1, EP3, and FP receptors and their vasoconstrictor functions in newborn pigs; the EP2 receptor was not significantly influenced by ontogenic alterations in prostaglandin levels. PMID- 9727413 TI - Measuring oxygen tension in the anterior chamber of rabbits. AB - PURPOSE: Measuring the concentration of oxygen in the aqueous humor without penetrating the eye would provide a new dimension in understanding aqueous humor and corneal dynamics. In this study a preinvasive method was developed for determining the cameral oxygen concentration in anesthetized rabbits by measuring the excited-state lifetime of a phosphorescent dye. METHODS: A scanning ocular fluorometer was designed to excite phosphorescence with a brief flash of light and to measure the decay of luminescence for as long as 1000 microsec after excitation. The measurement window was scanned through the depth of the anterior chamber or fixed at the mid-anterior chamber. A depot of the phosphorescent dye Pd-uroporphyrin was injected into the vitreous of eight pigmented rabbits, and within a few days the dye was measurable in the anterior chamber. The excited state lifetime of this dye is inversely correlated to oxygen concentration and was calibrated by measuring the lifetime of dye in cuvettes equilibrated with oxygen-nitrogen mixtures. Oxygen tensions were determined from lifetimes measured in the open eye, under a polymethylmethacrylate (PMMA) contact lens, under two oxygen-permeable contact lenses, and immediately after lid closure. RESULTS: Oxygen tension in the mid-anterior chamber before placing a PMMA contact lens was 23 +/- 3 mm Hg (mean +/- SD; n = 6). After 20 minutes of PMMA lens wear, oxygen tension decreased to 4 +/- 2 mm Hg. When the focal diamond was scanned through the anterior chamber, oxygen tension was 24 +/- 5 mm Hg near the corneal endothelium and decreased to 17 +/- 8 mm Hg near the crystalline lens. Under the PMMA contact lens this gradient reversed: Oxygen tensions near the endothelium and lens were 3 +/- 2 mm Hg and 6 +/- 2 mm Hg, respectively. Lid closure for 10 minutes or longer decreased the mid-anterior chamber oxygen tension from 21 +/- 2 mm Hg (n = 19 measurements from seven animals) to 10 +/- 3 mm Hg (n = 15 measurements from five animals). CONCLUSIONS: Measuring excited-state lifetime of phosphorescent dyes in the anterior chamber provides a useful method for determining oxygen concentration in vivo, without penetrating the eye. Cameral oxygen tension under PMMA contact lenses are significantly lower than in the uncovered eye. The profile of oxygen tension through the anterior chamber suggests that oxygen is supplied transcorneally to the aqueous humor. PMID- 9727414 TI - Effect of nitroprusside on arteriolar constriction after retinal branch vein occlusion. AB - PURPOSE: The development of extended areas of nonperfused capillaries after branch vein occlusion (BVO) is correlated to the secondary constriction of the arteriole crossing the occluded area. The decrease in nitric oxide (NO) in tissue that occurs early after BVO accounts for the secondary arteriolar constriction. The present study shows that the administration of an NO donor can reverse the secondary arteriolar vasoconstriction observed after BVO. METHODS: Simultaneous preretinal NO profiles and arteriolar diameter measurements were performed in miniature pigs after experimental BVO. The effect of preretinal microinjections of the NO donor sodium nitroprusside (SNP) on the arteriolar diameter was studied. RESULTS: Significant arteriolar vasoconstriction (mean arteriolar diameter, 92.1% +/- 3.3% of control; n = 7; P = 7.4 x 10(-5)) and a simultaneous decrease in the preretinal NO concentration ([NO]) (preretinal [NO], 20% +/- 15.6% of control; n = 5; P = 0.0003) were observed 4 hours after BVO. Microinjection of the NO donor SNP (1 mM applied by puffer) near the constricted retinal arteriole caused a segmental, reversible arteriolar dilation that reached its maximum 20 minutes after the injection (mean arteriolar diameter; 110.8% +/- 7.5% of control; n = 6; P = 0.02) and was completely reversed 60 minutes later (n = 6). CONCLUSIONS: Local administration of NO donors may contribute to the restoration of the retinal arteriolar blood flow after BVO and thus may improve the supply of oxygen and nutrients to the injured tissue. PMID- 9727415 TI - Clinical and histopathologic features of canine oxygen-induced proliferative retinopathy. AB - PURPOSE: In previous studies the morphologic features of the acute vaso obliterative and vasoproliferative stages of oxygen-induced retinopathy (OIR) were quantified and described in the dog model of retinopathy of prematurity (ROP). In the present study the sequelae of these events were examined using fluorescein angiography and histologic, enzyme, and immunohistochemical techniques. METHODS: Thirty newborn animals were exposed to 95% to 100% oxygen for 4 days and returned to room air until they were 22 to 45 days of age. Before death some animals were anesthetized, and fluorescein angiography was performed. Retina and vitreous from some animals were processed for adenosine diphosphatase (ADPase) flat-embedding. In other cases, eyes were prepared for full-thickness eyewall sectioning or frozen for histochemical analysis. RESULTS: Fluorescein angiography, funduscopic examination, and ADPase preparations showed dilated and tortuous retinal vessels, pigmentary changes, incomplete vascularization of peripheral retina, vitreous hemorrhage, and persistence of massive intravitreal neovascularization. Full-thickness eyewall sections showed tractional retinal folds, tented intravitreal vascularized membranes, and vitreous synchysis. Immunohistochemical analysis showed inner retinal astrogliosis. Enzyme histochemistry showed high alpha glycerophosphate dehydrogenase activity in poorly differentiated neovascular formations and low activity in formations with mature pericytes and endothelial cells. CONCLUSIONS: End-stage OIR in the neonatal dog shares many features with the chronic human disease. These results provide additional support for the use of this model in experimental studies of ROP. PMID- 9727416 TI - Diabetic-like retinopathy: early and late intervention therapies in galactose-fed rats. AB - PURPOSE: To determine whether the diabetic-like thickening of retinal capillary basement membrane (RCBM) that develops in the galactose-fed rat model of diabetic ocular complications could be halted or ameliorated after 4 or 8 months of galactosemia by treatment with ARI-509, a potent new aldose reductase inhibitor (ARI), or by withdrawal of the galactose diet. METHODS: Weanling female Sprague Dawley rats were randomized into eight groups and fed laboratory chow plus 50% starch, control group (CON); 50% D-galactose, galactose-fed group (GAL); 50% D galactose with ARI-509 at 25 mg/kg or 10 mg/kg body wt per day, high-dose prevention group (HDP) and low-dose prevention group (LDP), respectively; 50% D galactose for 4 or 8 months and then intervention by addition of ARI-509 (25 mg/kg body wt per day), 4-month intervention group (4IN) and 8-month intervention group (8IN), respectively; or 50% D-galactose for 4 or 8 months and then intervention by withdrawing galactose and replacing it with the 50% starch diet, 4-month galactose withdrawal group (4GW) and 8-month galactose withdrawal group (8GW), respectively. After 4, 8, 16, and 24 months of the experimental diets, the levels of carbohydrates in tissues and the extent of RCBM thickening in capillaries of the outer plexiform layer were determined in all groups. RESULTS: Retinal polyol was reduced by 95% in all ARI-treated groups and by 100% in the 4GW and 8GW groups after withdrawal of the galactose. The mean RCBM thickness increased rapidly in GAL rats, becoming almost two times greater (189 +/- 9.4 nm) than in CON rats (103 +/- 3.4 nm) by 24 months. Treatment with ARI-509 in high and low doses (HDP, LDP) initiated with the introduction of the galactose diet significantly prevented RCBM thickening at all time points (P < 0.05). In contrast, intervention by withdrawing galactose from the diet or by adding the high dose of ARI-509 had no significant effect (P < 0.05) on RCBM thickening until the 24-month time point (4IN, 166 +/- 10.3 nm; 8IN, 161 +/- 8.2 nm; 4GW, 136 +/- 5.1 nm; 8GW, 163 +/- 9.6 nm). CONCLUSIONS: Both early and late interventions decreased RCBM thickening compared with that in untreated GAL rats. The decreased thickening, however, was not evident until 16 to 20 months after the intervention. Because RCBM thickening is one of the earliest changes in diabetic and galactosemic retinopathy, the findings suggest that RCBM thickening and possibly subsequent retinal lesions are caused by early biochemical alterations induced by the galactose diet that are not readily reversed. The delayed response to therapy is consistent with that observed in the Diabetes Control and Complications Trial. The cumulative evidence indicates that intervention should begin as early after onset of diabetes as possible, and long follow-up periods should be used to evaluate efficacy. PMID- 9727417 TI - Measurement of adenosine concentration in aqueous and vitreous. AB - PURPOSE: The release of adenosine by the ischemic retina may be an initial signal in the development of ischemic macular edema and neovascularization. The levels of adenosine have never been quantified in ocular fluids. In this study, a technique was developed for in vivo measurement of the concentration of adenosine in aqueous and vitreous. METHODS: Aqueous and vitreous samples were obtained from bovine eyes after death and from live porcine eyes with the subject under general anesthesia. Samples from live eyes were immediately incubated in the sampling syringe with pentoxifylline, erythro-9-(2-hydroxy-3-nonyl) adenine, and dipyridamole to prevent synthesis or degradation of adenosine during the collection procedure, filtered, and flash-frozen in liquid nitrogen. All samples were then filtered and purified on phenylboronate agarose columns and incubated with chloroacetaldehyde to convert the adenosine present in the sample to the fluorescent derivative 1,N6-ethenoadenosine. The 1,N6-ethenoadenosine was separated by high-pressure liquid chromatography and then measured by fluorometry. RESULTS: Levels of adenosine as low as 0.5 pmole could be detected with this procedure, compared with 20 pmoles by UV detection. By using this technique to measure adenosine levels in the eyes of normal weanling domestic pigs, it was determined that the adenosine concentration in the aqueous was 321.3 +/- 164.9 nM and in the vitreous was 210.8 +/- 41.5 nM. CONCLUSIONS: The conversion of adenine-containing compounds to fluorescent 1,N6-etheno derivatives offers analytical advantages of selectivity and sensitivity for the quantitative determination of these compounds, with the fluorometric detection providing substantially greater sensitivity than direct detection by UV absorption. The levels obtained in vivo from anesthetized but otherwise healthy pigs presumably reflected basal aqueous and vitreous adenosine levels under the described conditions. This method should be useful in investigating more directly the role of adenosine in models of retinal or ocular ischemia in vivo and in measuring adenosine levels in vitreous or aqueous samples from human patients. PMID- 9727418 TI - Severe corneal dystrophy phenotype caused by homozygous R124H keratoepithelin mutations. AB - PURPOSE: To determine the mutational status of the beta ig-h3 gene in five patients from four Japanese families affected with an unusual, severe form of corneal dystrophy. In these five cases, the corneas were remarkable for confluent round opacities in the superficial stromal layer. The beta ig-h3 gene coding for keratoepithelin was recently identified as the gene responsible for 5q-linked autosomal dominant corneal dystrophies. METHODS: Genomic DNA was isolated from leukocytes of five patients with the severe form of corneal dystrophy. To screen for point mutations, exons of the beta ig-h3 gene were amplified by polymerase chain reaction and were analyzed with the single-strand conformational polymorphism technique. Subsequently, the mutations were identified by a direct sequencing method and restriction enzyme digestion analysis. RESULTS: All five patients with the severe form of corneal dystrophy had homozygous R124H keratoepithelin mutations. Histopathologic examinations of the corneas obtained from two patients with the severe form showed granular, rod-shaped deposits. CONCLUSIONS: The severe phenotype was a pathologic variant of granular corneal dystrophy (GCD). All five patients had homozygous R124H keratoepithelin mutations. The R124H keratoepithelin mutation is the same mutation recently reported to be responsible for Avellino corneal dystrophy. The homozygous R124H keratoepithelin mutations are the cause of the severe variant of GCD characterized by juvenile-onset and confluent superficial opacity. PMID- 9727419 TI - Cytokine expression during orthotopic corneal allograft rejection in mice. AB - PURPOSE: The acquisition of cell-mediated immunity against donor antigens has been shown to be associated with rejection of orthotopic corneal allografts, but the mechanisms that cause corneal allograft destruction in grafted tissue remain obscure. To determine which T-cell subsets infiltrate graft tissue and cause graft rejection, cytokine expression was examined in corneal tissue after orthotopic corneal allograft. METHODS: BALB/c mice received orthotopic corneal allografts from either syngeneic BALB/c or allogeneic C57BL/6 donors. At 1 or 4 weeks after grafting, the mice were euthanatized, and their corneas were removed. Corneal tissue was frozen, homogenized, and placed in phosphate-buffered saline (PBS). Each sample consisted of five corneas in 500 ml PBS. After centrifugation, the supernatant was collected, and the concentration of the following cytokines was measured by enzyme-linked immunosorbent assay: interleukin (IL)-1alpha, IL-2, IL-4, IL-10, interferon (IFN)-gamma, and tumor necrosis factor (TNF)-alpha. RESULTS: Significantly increased amounts of proinflammatory cytokines (IL-1alpha and TNF-alpha) were detected in supernatants from all grafted corneas (both syngeneic and allogeneic) at 1 week after grafting. At 4 weeks after grafting, supernatants from normal corneas, corneas with syngeneic grafts, and corneas with accepted corneal allografts contained undetectable amounts of IL-2 and IFN-gamma, whereas supernatants from corneas with rejected corneal allografts contained significant amounts of IL-2 and IFN-gamma. There were no significant differences in the amounts of IL-4 or IL-10 among all samples. Histologic examination confirmed the expression of IL-2 and IFN-gamma in rejected corneal allografts. CONCLUSIONS: Because IL-2 and IFN-gamma are secreted primarily by T-helper type 1 (Th 1) cells, whereas IL-4 and IL-10 are secreted by T-helper type 2 (Th 2) cells, these results indicate that Th 1-type cytokines, rather than Th 2-type cytokines, contribute to the rejection of orthotopic corneal allografts in graft tissue. PMID- 9727420 TI - Decorin and biglycan of normal and pathologic human corneas. AB - PURPOSE: Corneas with scars and certain chronic pathologic conditions contain highly sulfated dermatan sulfate, but little is known of the core proteins that carry these atypical glycosaminoglycans. In this study the proteoglycan proteins attached to dermatan sulfate in normal and pathologic human corneas were examined to identify primary genes involved in the pathobiology of corneal scarring. METHODS: Proteoglycans from human corneas with chronic edema, bullous keratopathy, and keratoconus and from normal corneas were analyzed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), quantitative immunoblotting, and immunohistology with peptide antibodies to decorin and biglycan. RESULTS: Proteoglycans from pathologic corneas exhibit increased size heterogeneity and binding of the cationic dye alcian blue compared with those in normal corneas. Decorin and biglycan extracted from normal and diseased corneas exhibited similar molecular size distribution patterns. In approximately half of the pathologic corneas, the level of biglycan was elevated an average of seven times above normal, and decorin was elevated approximately three times above normal. The increases were associated with highly charged molecular forms of decorin and biglycan, indicating modification of the proteins with dermatan sulfate chains of increased sulfation. Immunostaining of corneal sections showed an abnormal stromal localization of biglycan in pathologic corneas. CONCLUSIONS: The increased dermatan sulfate associated with chronic corneal pathologic conditions results from stromal accumulation of decorin and particularly of biglycan in the affected corneas. These proteins bear dermatan sulfate chains with increased sulfation compared with normal stromal proteoglycans. PMID- 9727421 TI - Quantitative evaluation of papilledema in pseudotumor cerebri. AB - PURPOSE: To determine the feasibility of adapting confocal scanning laser (CSL) tomography of the optic disc for quantitative evaluation of papilledema in pseudotumor cerebri (PTC). METHODS: Confocal scanning laser tomography of the optic disc was performed in 11 patients with diagnosed PTC and 12 visually normal control subjects of similar age. In five patients with active papilledema, CSL tomography was performed serially over several months. To quantify optic disc characteristics, surface topography was measured in 0.1-mm steps along the horizontal and vertical meridians and four oblique meridians. Best fit polynomial functions, describing surface topography along each meridian, were derived using linear regression analysis. RESULTS: Third-order polynomials provided excellent fits (significantly better than the second-order functions) to the surface topography for all meridians in the control subjects and patients with PTC. In control subjects and PTC patients an asymmetry in the slope of the optic disc contours was evident along the horizontal but not the vertical meridian. In patients with active papilledema a significant elevation of the center of the disc was accompanied by a change in overall surface topography. Each of the PTC patients followed up serially had a pronounced posterior deformation of the disc (i.e., a reduction in papilledema) that was initially apparent in the temporal meridian and did not proceed uniformly across all meridians. CONCLUSIONS: Confocal scanning laser tomography can quantify the magnitude and monitor the resolution of papilledema in PTC. Studies of optic nerve head topography may provide further insight into optic nerve compliance with elevated intracranial pressure. PMID- 9727422 TI - Ultrastructural changes in rabbit ciliary body after extraocular mitomycin C. AB - PURPOSE: To study the ultrastructural changes in ciliary body epithelium of the rabbit eye after subconjunctival injections of mitomycin C. METHODS: One eye of six New Zealand white rabbits was given a subconjunctival injection at the 12 o'clock position with 0.005, 0.02, 0.08, 0.1, 0.12, or 0.16 mg mitomycin C. The fellow eye was given a subconjunctival injection of balanced salt solution. Two weeks after treatment, the eyes were enucleated, and the ciliary body was exposed and submerged in fresh 4% paraformaldehyde/2% glutaraldehyde in 0.1 M phosphate buffer, pH 7.4, at 4 degrees C. Electron microscopy of the ciliary body was performed at two sites: the injection site (12-o'clock position) and 180 degrees away (6-o'clock position). RESULTS: At dosages of 0.1 mg and higher, ciliary body epithelial cells beneath the injection site were thinned. There were vacuoles and expansion of intracellular and intercellular spaces. Plasma membrane infoldings were disrupted, and the apical membrane was thinned. Mitochondria and nuclei were normal. Ciliary body epithelium at 6-o'clock position showed only mild architectural distortion of the plasma membrane infoldings. Eyes that received lower doses of mitomycin C (0.005 mg, 0.02 mg, and 0.08 mg) and balanced salt solution showed normal ciliary body epithelium at the injection site and 180 degrees away. CONCLUSIONS: Subconjunctival injection of mitomycin C in the rabbit produces dose-dependent localized ultrastructural changes of the ciliary body epithelium. PMID- 9727423 TI - Muller cell-specific autoantibodies in a patient with progressive loss of vision. AB - PURPOSE: To study the specificity of circulating retinal autoantibodies in a patient with progressive loss of vision resembling cancer-associated retinopathy in the absence of systemic malignancy. METHODS: Patient's serum was tested for the presence of antiretinal antibodies by Western blot analysis, and immunohistochemical analysis was performed on cryosections of rat retina and on cultured rat retinal Muller cells. RESULTS: Western blot analysis revealed that the serum contained a high titer of autoantibodies against a 35-kDa retinal antigen. A protein of similar molecular weight and antigenicity has been found to be present in protein extracts of bovine, rat, and fish retina. Immunohistochemical analysis suggested that the autoantibodies did not localize to retinal neural cells, as reported for several other putative autoimmune retinal disorders, but rather to the retinal Muller cells. This cell type specificity could be confirmed using purified cultured retinal Muller cells. CONCLUSIONS: These results suggest that an autoimmune response directed against the retinal Muller cells, in the absence of overt systemic malignancy, may lead to a slow-developing visual deterioration. PMID- 9727424 TI - Angiosomes of the leg: anatomic study and clinical implications. AB - In 1987, Taylor and Palmer introduced the angiosome concept. This anatomical study defined the three-dimensional vascular territories supplied by source arteries and veins to each tissue layer between the skin and the bone. This report, however, was an overview investigation and did not study each region of the body in fine detail. In 1996, Inoue and Taylor studied the angiosomes of the forearm in much greater detail. They showed, among other findings, that the zone between the angiosomes, formed by reduced caliber (choke) vessels or similar caliber (true) anastomotic arteries, occurred usually within tissues, especially the muscles, not between them. This study focuses on the same region in the lower limb to draw a comparison and to fill certain voids in our knowledge--the leg. Twelve lower limbs from fresh cadavers were investigated over a 2-year period after perfusing each with a mixture containing radio-opaque lead oxide. The anatomy of the arterial supply to the skin, the muscles, and the periosteum of the bones of the leg was examined. The contribution to each tissue was defined by dissection, by metal clip tagging of vessels, by radiography, and by mapping the branches with colored pins, coded to match the respective source arteries. A subtraction technique was used to study the muscles whereby the bones of the limb were replaced with radiolucent balloons to obtain an unobscured picture of the vasculature of the leg. The muscles were then segregated one by one from the muscle mass and x-rayed again. Next, cross-section studies were made in two legs to complete the three-dimensional picture, tracing the branches from the source arteries to each layer. Finally, the contribution to each tissue from the popliteal, sural, anterior tibial, posterior tibial, and peroneal vessels were color coded to match these source arteries, thus defining the angiosomes of the leg. Results, as in the forearm, showed that in most cases the connections between adjacent angiosomes occurred within tissues, not between them. The skin, the bones, and most muscles received branches from two or more angiosomes, thus revealing one of the important anastomotic pathways through which the circulation is reconstituted when a source artery is interrupted by disease or trauma. Notably, however, the muscles of the anterior compartment of the leg were supplied from one angiosome. This finding, coupled with the anatomy of the rigid fascial compartments of the leg, helps explain the variable clinical pictures and syndromes seen in cases in which the circulation is compromised or interrupted. Finally, this anatomical study adds further information to help design or redesign flaps in the leg for local or free transfer. Similarly, the information reveals the pathways through which the supply to the remaining tissues is reconstituted when one of the source arteries is harvested with a free flap, especially when multiple tissues are included in the transplant. PMID- 9727425 TI - Nonsyndromal craniosynostosis: longitudinal outcome following cranio-orbital reconstruction in infancy. AB - This paper presents a prospective longitudinal outcome study on patients with nonsyndromal craniosynostosis who were treated with the contemporary craniofacial surgical techniques of suture release, cranial decompression, and cranial and orbital reconstruction and reshaping in infancy. Diagnosis, surgical treatment, and long-term results and complications are reviewed. Preoperative and long-term postoperative intracranial volumes in these patients were evaluated and compared with age and gender match controls throughout the period of the study. From July 1, 1990, to July 1, 1994, 25 patients with isolated nonsyndromal craniosynostosis underwent surgery of the deformity. Eight patients were excluded from the study based on incomplete postoperative computed tomography (CT) records. Of the 17 patients with long-term computerized records, 11 were boys and 6 were girls. The nonsyndromal craniosynostosis patients in this study include six with bilateral coronal craniosynostosis, six with unilateral coronal craniosynostosis, four with sagittal craniosynostosis, and one with metopic craniosynostosis. The average age at the time of surgery for all patients was 9 months, and the average age at the time of the latest follow-up CT scan for all patients in the study was 3.5 years. There were no perioperative complications in this series of patients including no bleeding, no infection, no wound healing complications, and no mortality. Bony fixation included a combination of wire osteosynthesis and rigid microfixation. All patients had only one surgical procedure for the correction of their deformity. Evaluation of both preoperative and long-term postoperative intracranial volume measurements in this series of patients revealed that these volume measurements were comparable with the gender match control groups at all ages throughout the study. The significance of these findings for this longitudinal outcome study is discussed. PMID- 9727426 TI - Transcranial correction of orbital neurofibromatosis. AB - Neurofibromatosis is a systemic disease that often produces striking disfigurement. Orbital manifestations are common and include sphenoid dysplasia with or without infiltration of the periorbital soft tissues. The resultant deficiency of the posterolateral orbital wall may lead to protrusion of the temporal lobe into the orbit, displacement of the globe, and pulsatile exophthalmos. Treatment at our unit has consisted of transcranial orbital reconstruction with bone grafts and periorbital soft-tissue correction. Observation of complete bone graft resorption in one patient prompted an assessment of the Australian Craniofacial Unit's experience with particular attention paid to the stability of operative correction. Of 36 patients with head and neck neurofibromatosis treated during the period from 1981 to 1995, 14 patients underwent transcranial correction of orbital deformities secondary to sphenoid dysplasia. The treatment and outcomes of this transcranial group are reviewed. The most notable finding was that of recurrent globe pulsation in four patients following initial resolution. Computed tomography scans have documented partial to complete bone graft resorption in three of these patients. Titanium mesh is now being utilized to provide a more durable reconstruction. PMID- 9727427 TI - Arteriovenous malformations of the head and neck: natural history and management. AB - This is a retrospective review of 81 patients with extracranial arteriovenous malformation of the head and neck who presented to the Vascular Anomalies Program in Boston over the last 20 years. This study focused on the natural history and effectiveness of treatment. The male to female ratio was 1:1.5. Arteriovenous malformations occur in anatomic patterns. Sixty-nine percent occurred in the midface, 14 percent in the upper third of the face, and 17 percent in the lower third. The most common sites were cheek (31 percent), ear (16 percent), nose (11 percent), and forehead (10 percent). A vascular anomaly was apparent at birth in 59 percent of patients (82 percent in men, 44 percent in women). Ten percent of patients noted onset in childhood, 10 percent in adolescence, and 21 percent in adulthood. Eight patients first noted the malformation at puberty, and six others experienced exacerbation during puberty. Fifteen women noted appearance or expansion of the malformation during pregnancy. Bony involvement occurred in 22 patients, most commonly in the maxilla and mandible. In seven patients, the bone was the primary site; in 15 other patients, the bone was involved secondarily. Arteriovenous malformations were categorized according to Schobinger clinical staging: 27 percent in stage I (quiescence), 38 percent in stage II (expansion), and 38 percent in stage III (destruction). There was a single patient with stage IV malformation (decompensation). Stage I lesions remained stable for long periods. Expansion (stage II) was usually followed by pain, bleeding, and ulceration (stage III). Once present, these symptoms and signs inevitably progressed until the malformation was resected. Resection margins were best determined intraoperatively by the bleeding pattern of the incised tissue and by Doppler. Subtotal excision or proximal ligation frequently resulted in rapid progression of the arteriovenous malformation. The overall cure rate was 60 percent, defined as radiographic absence of arteriovenous malformation. Cure rate for small malformations was 69 percent with excision only and 62 percent for extensive malformations with combined embolization-resection. The cure rate was 75 percent for stage I, 67 percent for stage II, and 48 percent for stage III malformations. Outcome was not affected significantly by age at treatment, sex, Schobinger stage, or treatment method. Mean follow-up was 4.6 years. PMID- 9727428 TI - Clinical implications of the velopharyngeal blood supply: a fresh cadaveric study. AB - The aim of this investigation was to examine the blood supply of the normal velopharyngeal musculature and its clinical implications. Detailed dissections were performed on each side of five fresh human adult cadaveric head and neck specimens (n = 10) following carotid artery injection with liquid neoprene latex stained with green pigment. The vascular network of the soft palate was situated within its glandular layer. The velopharyngeal muscles were supplied by the following four branches of the external carotid artery: (1) ascending palatine branch of the facial artery, which supplied the palatoglossus, palatopharyngeus, musculus uvulae, and the intravelar part of the levator veli palatini; (2) ascending pharyngeal artery, which supplied the superior constrictor; (3) the previously undescribed recurrent pharyngeal artery, which supplied the extravelar part of the levator veli palatini; and (4) maxillary artery, which supplied the tensor veli palatini. All muscles except the musculus uvulae had at least a dual blood supply. Analysis of this vascular anatomy suggests that (1) the overall generous blood supply of the velum allows it to tolerate the dissection performed during intravelar veloplasty and the Furlow double opposing Z-plasty; (2) dissection around the hamulus, along the medial pterygoid plate, and in the space of Ernst should be performed carefully to avoid damage to the ascending palatine artery, ascending pharyngeal, and recurrent pharyngeal arteries; (3) vertical pharyngeal flaps are random pattern in nature; and (4) the posterior tonsilar pillar flaps of the sphincter pharyngoplasty are adequately supplied by the hamular branch of the ascending palatine artery. PMID- 9727429 TI - A fiberscopic analysis of velopharyngeal movement before and after primary palatoplasty in cleft palate infants. AB - There have been few studies done on the abnormal function of velopharyngeal muscles in unrepaired cleft palate infants. To examine and assess velopharyngeal movement before primary palatoplasty offers supposedly any valuable information for the successful operation and the restoration of excellent velopharyngeal function. We designed to investigate and analyze velopharyngeal movement before and after primary palatoplasty in 26 cleft palate infants with a fine nasopharyngeal fiberscope. We found three different patterns of velopharyngeal movement in unrepaired cleft palate infants when crying or strangulation reflex occurred: (1) posterior movement type (10 cases, 38.5 percent), where the soft palates moved only posteriorly and cephalically and did not move medially; (2) medial movement type (10 cases, 38.5 percent), where the soft palates moved only medially and did not move posteriorly or cephalically; and (3) posteromedial movement type (6 cases, 23.0 percent), where the soft palates moved both posteriorly and cephalically as well as medially. Postoperative velopharyngeal closure was classified into three patterns: (1) the soft palate type, in which the soft palate mainly operates; (2) the lateral wall type, in which compensational medial movement of the lateral pharyngeal wall is mainly observed; and (3) the mixed type, in which both the soft palate and the lateral pharyngeal wall operate. Also, we demonstrated a close relationship between velopharyngeal movement before and after primary palatoplasty in cleft palate infants. In total, 10 of 16 cleft palate infants with the posterior movement type or posteromedial movement type, in which posterior movement of the soft palates was observed before primary palatoplasty, postoperatively showed the soft palate type of velopharyngeal closure. On the other hand, only 2 of 10 cleft palate infants with the medial movement type, in which the soft palates did not move posteriorly but medially before primary palatoplasty, postoperatively showed the soft palate type of velopharyngeal closure. The Fisher's exact probability test clarified that cleft palate infants with the posterior movement type or posteromedial movement type were more likely to show postoperatively the soft palate type of the velopharyngeal closure compared with those with the medial movement type (p = 0.051). This is the first trial to examine velopharyngeal movement in unrepaired cleft palate infants. Our findings indicate the probability that velopharyngeal closure mechanism in repaired cleft palate infants is able to be predicted by velopharyngeal movement behavior before primary palatoplasty. Next, we must clarify a correlation between preoperative velopharyngeal movement and postoperative velopharyngeal function and speech outcome. PMID- 9727430 TI - Speech outcome and maxillary growth in patients with unilateral complete cleft lip/palate operated on at 6 versus 12 months of age. AB - A prospective study of speech outcome and maxillofacial growth was carried out in cleft palate patients. Seventy-six cleft palate patients were randomly selected for the study group; 41 patients were operated on at 12 months of age, and 35 patients were operated on at 6 months of age. All patients were followed until they were 4 years of age. All patients underwent a complete speech evaluation, videonasopharyngoscopy, videofluoroscopy, and maxillofacial assessment. The rate of velopharyngeal insufficiency did not differ between the two groups (17 to 19 percent; p > 0.05). However, phonologic development was significantly better (p < 0.05) in the patients operated on at 6 months of age. Furthermore, none of the patients operated on at 6 months of age showed compensatory articulation disorder. In contrast, 62 percent of the patients with postoperative velopharyngeal insufficiency operated on at 12 months of age showed compensatory articulation disorder (p < 0.05). Maxillofacial assessment showed that there were non-significant differences (p > 0.05) in maxillofacial growth in both groups of patients. All patients showed similar degrees of maxillary collapse (p > 0.05). The results of this study suggest that cleft palate repair performed at 6 months of age significantly enhances speech outcome and prevents compensatory articulation disorder. PMID- 9727431 TI - Primary insertion of osseointegrated dental implants into fibula osteoseptocutaneous free flap for mandible reconstruction. AB - Twelve patients with segmental mandibular defects were reconstructed with fibula osteoseptocutaneous flaps and simultaneous placement of osseointegrated implants. Decision to perform this procedure was based on the facts that all patients had benign diseases, did not require postoperative radiotherapy, were in good general and oral conditions, and were psychologically motivated. A total of 34 fixtures was inserted in the first stage. Eight patients underwent second stage surgery, which consisted of connection of the implant abutments to the fixtures and the use of palatal mucosal grafts around the implants. Final dental prostheses were fixed 1 month later in seven patients, at this time. All flaps survived after surgery, and no implant failure was observed after a mean follow-up period of 25 months. Only one fixture was not used during the subsequent stage and was left as a sleeper. Fixed dental prostheses were used in five patients and removable overlay prostheses in the other two. Chewing function was recovered between 4 and 6 weeks after the start using the definitive dental prosthesis. In contrast to previous results, we conclude that excellent results can be achieved when this combined procedure is used in carefully selected patients. In addition, it is confirmed that the fibula osteoseptocutaneous flap is a versatile, reliable composite tissue that facilitates primary placement of osseointegrated dental implants during mandible reconstruction, thus allowing full oral rehabilitation in a shorter period of time. PMID- 9727432 TI - Decision-analysis methodology in the work-up of women with suspected silicone breast implant rupture. AB - Despite numerous studies advocating ultrasonography and magnetic resonance imaging (MRI) in the evaluation of women with possible silicone breast implant rupture, an appropriate algorithm has not been published for the optimal use of these tests. To derive a diagnostic algorithm using ultrasonography and MRI, we applied a decision-analytic model using Bayes' theorem to calculate the probabilities of implant rupture for three representative patient characteristics. A Medline search was conducted to identify literature related to the diagnosis of silicone breast implant rupture using ultrasonography and MRI since 1994. Also examined were case series of implant rupture to obtain rupture prevalence in cases in which rupture was dependent on patient presentation (symptomatic versus asymptomatic) and dependent on implant age. Test characteristics (sensitivity and specificity) and implant rupture prevalence are used to calculate the probability of rupture by using Bayes' theorem. These probabilities are derived for three patient categories: (1) asymptomatic, (2) symptomatic with implant age < or = 10 years, and (3) symptomatic with implant age >10 years. In asymptomatic patients, the pretest rupture prevalence is 6.5 percent. If a screening ultrasonography shows no rupture, the probability of rupture drops to 2.2 percent. If ultrasonography shows rupture, the probability of true rupture increases to 37.8 percent. Removal of implants in this setting will result in a high probability of extracting normal implants. However, if MRI after the ultrasonography shows rupture, the probability of true rupture increases to 86 percent, which gives better assurance of removing true-ruptured implants. In "symptomatic" patients (i.e., breast asymmetry, capsular contracture) with implants < or = 10 years old, the prevalence of rupture is estimated at 31 percent. If ultrasonography shows no rupture, the probability of rupture drops to 16 percent. If ultrasonography shows rupture, the probability of true rupture is 79.7 percent, and this probability increases to 97.5 percent if a follow-up MRI also shows rupture. In symptomatic patients with implants >10 years old, the prevalence of rupture is estimated at 64 percent. If ultrasonography shows rupture, the probability of true rupture increases to 94 percent, and no further diagnostic work-up is necessary. In an asymptomatic patient who is worried about the integrity of her implants, ultrasonography should be used as an initial diagnostic test because of its lower cost. If ultrasonography shows no rupture, no further work-up is necessary. If ultrasonography shows rupture, the low probability (37.8 percent) of true rupture requires a confirmatory test using MRI. In "symptomatic" patients, the high prevalence of rupture markedly raises the posttest probability of rupture for positive ultrasonography findings. Particularly in "symptomatic" patients with implants >10 years old, the high posttest probability of rupture (94 percent) with a positive ultrasonography obviates the need for any further diagnostic testing. This diagnostic algorithm will assist plastic surgeons in counseling women who are worried about the integrity of their silicone breast implants. PMID- 9727433 TI - Carcinoma of the male breast: an underestimated killer. AB - Male breast carcinoma represents only 1 percent of all breast cancers. Despite the rarity of this neoplasia, the malignancy is often very aggressive, mostly because of delayed diagnosis. Consequently, large excisions are usually needed, and subsequent reconstruction is performed by means of fasciocutaneous or myocutaneous local or distant flaps, depending on the stage of the neoplasia. The signs, symptoms, prognosis, and reconstructive methods of such an unusual form of breast cancer are discussed in this article. PMID- 9727434 TI - Normal cutaneous sensibility of the breast. AB - A total of 150 healthy women were studied to determine normal values for breast sensibility and to investigate the influence of breast size and ptosis on breast sensation. Cutaneous pressure thresholds were evaluated bilaterally in six areas including the nipple, the areola, and the skin of the breast using the Semmes Weinstein monofilaments. We found that the skin of the superior quadrant was the most sensitive part of the breast, the areola was less sensitive, and the nipple was the least sensitive part. The cutaneous sensibility of all tested areas decreased significantly with increasing breast size and increasing breast ptosis. The nipple was less sensitive in women who had a previous pregnancy. Age, smoking history, or hormonal contraception had no significant influence on breast sensation. The study shows that the Semmes-Weinstein test is an adequate method for assessing sensation in the breast. PMID- 9727435 TI - Sweat gland carcinoma: a clinicopathologic analysis of an expanded series in a single institution. AB - Primary adenocarcinoma of sweat glands is a rare tumor; approximately 220 cases have been reported in the last 30 years. We reviewed the charts of patients with primary diagnosis of this tumor treated at the Mayo Clinic between 1935 and 1995. We included only cases with initial histology slides available for re examination. Tumors were classified into five recognizable histologic patterns (solid, ductal, mucinous, microcystic adnexal, and adenocystic carcinoma) and graded by the Broder system. Statistical analysis consisted of Kaplan-Meier product limit method and Cox multiple regression test. In total, 55 patients were identified, and age ranged from 13 to 85 years (mean 59 years). Thirty-six patients (65 percent) presented to the Mayo Clinic for initial treatment; all except one had disease limited to the primary site. Microcystic adnexal carcinoma was the most frequent type, and more than 50 percent were grade 2 tumors. Among these 36 patients, 4 had some type of recurrence. Patients who developed metastasis had a high-grade tumor in the initial biopsy. Nineteen patients were referred with recurrence; 13 had local recurrence, 4 had regional diseases, and 2 had distant metastases. The histologic distribution showed 47 percent solid tumors, and 37 percent of them were grade 3. Multiple regression analysis did not show a difference in recurrence or survival when gender, age, tumor location, or histologic pattern was evaluated. In addition, there was no difference in the outcome between wide surgical resection and micrographic surgery. The only predictive factor for distant metastases and/or death (p < 0.003) was histologic grade. Overall 10-year survival rate was 86 and 60 percent for primary and referred patients, respectively. We conclude that histologic diagnosis of sweat gland carcinoma must be complemented by clinical examination to evaluate metastases. Clinical behavior depends on the histologic type of tumor, degree of differentiation, and clinical stage. On recurrence, the likelihood of further recurrences and mortality increases dramatically. Aggressive initial local ablation with tumor-free margins is recommended. In high-grade tumors, prophylactic regional lymph node dissection may further characterize tumor aggressiveness and may justify adjuvant radiotherapy as part of the primary treatment. PMID- 9727436 TI - A prospective study of microvascular free-flap surgery and outcome. AB - Over a 6-month period, 23 members of the International Microvascular Research Group participated in a prospective survey of their microvascular free-flap practice. Data were recorded with each case for 60 variables covering patient characteristics, surgical technique, pharmacologic treatment, and postoperative outcome. A total of 493 free flaps were reported with a representative demographic distribution for age, sex, indications for surgery, risk factors, flap type, surgical technique, and pharmacologic intervention. Mixed effects logistic regression modeling was used to determine predictors of flap failure and associated complications. The overall incidence of flap failure was 4.1 percent (20 of 493). Reconstruction of an irradiated recipient site and the use of a skin grafted muscle flap were the only statistically significant predictors of flap failure, with increased odds of failure of 4.2 (p = 0.01) and 11.1 (p = 0.03), respectively. A postoperative thrombosis requiring re-exploration surgery occurred in 9.9 percent of the flaps. The incidence of this complication was significantly higher when the flap was transferred to a chronic wound and when vein grafts were needed, with increased odds of failure of 2.9 (p = 0.02) and 2.5 (p = 0.02), respectively. There was a lower incidence of postoperative thrombosis when rectus/transverse rectus abdominis muscle (TRAM) flaps were used, where odds of failure decreased by 0.36 (p = 0.04), and when subcutaneous heparin was administered in the postoperative period, where odds decreased by 0.27 (p = 0.04). There was an overall 69-percent salvage rate for flaps identified with a postoperative thrombosis. Intraoperative thrombosis occurred in 41 cases (8.3 percent) and was observed more frequently in myocutaneous flaps or when vein grafts were needed (5.5 and 5.0 greater odds, respectively; p < 0.001) but was not associated with higher flap failure (2 of 41 cases; 4.9-percent failure rate). The incidence of a hematoma and/or hemorrhage was increased in obese patients and when vein grafts were needed [2.7 (p = 0.02) and 2.6 (p = 0.03) greater odds, respectively], whereas this complication was significantly decreased in muscle flaps (myocutaneous or skin-grafted muscle), in tobacco users, when a heparinized solution was used for general wound irrigation, and when the attending surgeon performed the arterial anastomosis (in contrast to the resident or fellow on staff) (p < 0.05 for each factor). With the multivariable analysis, many factors were found not to have a significant effect on flap outcome, including the recipient site (e.g., head/neck, breast, lower limb, etc.); indications for surgery (trauma, cancer, etc.); flap transfer in extremes of age, smokers, or diabetics; arterial anastomosis with an end-to-end versus end to-side technique; irrigation of the vessel without or with heparin added to the irrigation solution; and a wide spectrum of antithrombotic drug therapies. These results present a current baseline for free-flap surgery to which future advances and improvements in technique and practice may be compared. PMID- 9727437 TI - A review of 716 consecutive free flaps for oncologic surgical defects: refinement in donor-site selection and technique. AB - Free-tissue transfer has become an important method for reconstructing complex oncologic surgical defects. This study is a retrospective review of a 10-year experience with 716 consecutive free flaps in 698 patients. Regional applications included the head and neck (69 percent), trunk and breast (14 percent), lower extremity (12 percent), and upper extremity (5 percent). Donor sites included the rectus abdominis (195), fibula (193), forearm (133), latissimus dorsi (69),jejunum (55), gluteus (28), scapula (26), and seven others (17). Microvascular anastomoses were performed to large-caliber recipient vessels using a continuous suture technique; end-to-end anastomoses were preferred (75 percent). Flaps were designed to avoid the need for vein grafts. Conventional postoperative flap monitoring methods were used. These included clinical observation supplemented by Doppler ultrasonography, surface temperature probes, and pin prick testing. Buried flaps were either evaluated with Doppler ultrasonography or not monitored. The overall success rate for free-flap reconstruction of oncologic surgical defects was 98 percent. Fifty-seven flaps (8 percent) were reexplored for either anastomotic or infectious problems. Reexplored flaps were salvaged in 40 cases (70 percent). Surviving flaps resulted in a healed wound and did not delay postoperative radiation or chemotherapy. The incidence of major and minor postoperative complications was 34 percent. The mean duration of hospitalization was 20 days, and the average cost was $40,224. The results of this study support the need for only seven donor sites to solve the majority (98 percent) of oncologic problems requiring microsurgical expertise. The evolution of preferred donor sites for specific regional applications is illustrated in this 10-year experience. Technical refinements have simplified performing the microsurgical anastomoses and essentially eliminated the need for vein grafts. Conventional monitoring has led to the rapid identification of vascular compromise and subsequent flap salvage in the majority of non-buried free flaps. PMID- 9727438 TI - The myoadipose flap: a new composite. AB - A prefabricated composite fat flap consisting of muscle woven into an anatomically distinct fat pad was studied in a rabbit model. In 17 rabbits, a 2 cm strip of latissimus dorsi was woven into the parascapular fat pad on one side, with the contralateral fat pad serving as a control. At 3 weeks, the endogenous blood supply of both the control and the experimental fat pads was isolated and ligated, and the composite fat/muscle flap was transferred to the chest wall. At 6 weeks, animals were killed, and flaps were analyzed for length, width, and weight; perfused with fluorescein or lead oxide; and examined histologically. Significant differences were found between the control and experimental fat pads with regard to weight and length. Experimental flaps were found to be perfused fully with fluorescein and lead oxide; control fat pads were found not to be perfused. The lead oxide group revealed extensive growth of blood vessels from the latissimus graft into the experimental fat pad. No vessels were visualized in the controls. Finally, sections of the control and experimental flaps were analyzed histologically. A preponderance of viable fat, with evidence of neovascularization, was found in experimental flaps, compared with the necrotic fat that characterized the controls. We conclude that prefabrication of a fat flap is possible and may have extensive application in various areas of plastic surgery. PMID- 9727439 TI - U-I flap. AB - This article describes the principle, vascular basis, and applications of a new flap derived from the dorsal hand skin. The principle is a pedicle-lengthening procedure that is to revert a "U"-shaped vascular structure into an "I"-shaped vascular structure. By using this principle, a dorsal metacarpal flap, comprising the second dorsal metacarpal artery, the dorsal carpal arch, the dorsal branch of the radial artery, and the first dorsal metacarpal artery, was developed. In this flap, the skin territory was placed over the first dorsal metacarpal artery. The flap was rotated around a web space-connecting vessel between the second dorsal metacarpal artery and the palmar arterial system. We used this flap for resurfacing large defects beyond the proximal interphalangeal joint and degloved fingers in six cases. In two cases, the flap was used as an innervated flap. In one case, the flap was used as a tenocutaneous flap with a segment of the extensor indicis proprius tendon. We found this flap to be useful in selected circumstances. PMID- 9727440 TI - Three-dimensional analysis and classification of arteries in the skin and subcutaneous adipofascial tissue by computer graphics imaging. AB - To develop new types of surgical flaps that utilize portions of the skin and subcutaneous tissue (e.g., a thin flap or an adipofascial flap), three dimensional investigation of the vasculature in the skin and subcutaneous tissue has been anticipated. In the present study, total-body arterial injection and three-dimensional imaging of the arteries by computer graphics were performed. The full-thickness skin and subcutaneous adipofascial tissue samples, which were obtained from fresh human cadavers injected with radio-opaque medium, were divided into three distinct layers. Angiograms of each layer were introduced into a personal computer to construct three-dimensional images. On a computer monitor, each artery was shown color-coded according to the three portions: the deep adipofascial layer, superficial adipofascial layer, and dermis. Three-dimensional computerized images of each artery in the skin and subcutaneous tissue revealed the components of each vascular plexus and permitted their classification into six types. The distribution of types in the body correlated with the tissue mobility of each area. Clinically, appreciation of the three-dimensional structure of the arteries allowed the development of several new kinds of flaps. PMID- 9727441 TI - Arterial vascular anatomy of the umbilicus. AB - The rare occurrence of umbilical necrosis after performance of a transverse rectus abdominis muscle (TRAM) flap prompted this investigation into the specific arterial anatomy of the umbilicus using multiple anatomic techniques. Sixteen fresh cadavers were studied by using dissection of blue latex-injected specimens, radiography of barium latex-injected specimens, and selective ink injection of individual perforators. It was discovered that the umbilicus receives arterial inflow by means of three distinct deep sources in addition to the subdermal plexus. These deep sources are (1) the right and left deep inferior epigastric arteries that each give off several small branches, and a large ascending branch, which courses between the muscle and the posterior rectus sheath passing directly to the umbilicus; (2) the ligamentum teres hepaticum; and (3) the median umbilical ligament. The clinical implications of this study are that the umbilicus should have robust arterial inflow if only one rectus muscle is removed, such as during a unilateral TRAM flap, because the contralateral side should still provide large direct vessels from the deep inferior epigastric arteries to the umbilicus. During bilateral TRAM elevation, all of the large arterial sources are removed from the umbilical inflow and circulation must depend on small vessels from the ligamentum teres and median umbilical ligament. Care should be taken in this latter clinical situation to preserve these sources of blood flow during umbilical flap creation. PMID- 9727442 TI - Results of 268 pressure sores in 158 patients managed jointly by plastic surgery and rehabilitation medicine. AB - Despite improvements in surgical repair of pressure sores, recurrence rates greater than 80 percent are reported, thus indicating that this difficult problem is not yet solved. Recurrence results in additional hospitalizations and increased medical expenses. Because associated general clinical and social issues are numerous for these patients, our physical medicine and rehabilitation colleagues are active participants in their perioperative medical care. In addition, the Department of Physical Medicine and Rehabilitation also directs a complete postreconstruction rehabilitation and education program for them. The results of surgically repaired pressure sores for patients managed in this collaborative fashion have not been previously reported. Pressure sore patients at the Harborview and University of Washington Medical Centers are evaluated by plastic surgery colleagues together with the Department of Physical Medicine and Rehabilitation. Patients believed to be candidates for complete postoperative rehabilitation are offered surgical repair and constitute this study cohort. Individuals who cannot cooperate with our protocol are treated nonoperatively and are not included in this study. A retrospective analysis of all 158 patients (mean age 34.5 years) operated on for 268 grade III and IV pressure sores between October of 1977 and December of 1989 was performed. Following surgical debridement and reconstruction, patients receive their principal medical care from the Department of Physical Medicine and Rehabilitation service while the Plastic Surgery Department manages the surgical wounds. Graduated patient mobilization is initiated in accord with a mutually agreed upon standardized protocol. New or primary sores numbered 174 (65 percent), and recurrent or secondary sores numbered 94 (35 percent). Mean patient follow-up was 3.7 years. The overall pressure sore recurrence rate (recurrence at the same site) was 19 percent, and the overall patient recurrence rate (previous patient developing a new sore) was 25 percent. Recurrence rates of 22 and 15 percent were noted for primary and secondary pressure sores, respectively. On most recent examination, 131 patients (83 percent) had intact pelvic and perineal skin. These results support a collaborative approach to the management of high-grade pressure sore patients. Our protocol of mutual patient evaluation followed by surgical reconstruction and postoperative rehabilitation yields notably low recurrence rates of both primary and secondary sores. In addition, the high percentage of patients who manifest long-term maintenance of skin integrity demonstrates the excellent education in personal skin and self-care that this approach provides. Not only do patients enjoy successful and durable reconstructive results, but additional hospitalizations and health care expenses implicit to pressure sore recurrence are consequently diminished. This collaborative clinical effort remains our standard of care. PMID- 9727443 TI - Division of the transverse carpal ligament and flexor tendon excursion: open and endoscopic carpal tunnel release. AB - Ten fresh cadaver upper extremities from 10 different subjects were used in this study of the effect of both open and endoscopic carpal tunnel release on flexor tendon excursion. The amount of excursion necessary to bring each finger from the fully extended to the fully flexed position with the fingertip just touching the palm was measured with the extremity mounted in a device that moved the wrist from extension through flexion. Endoscopic carpal tunnel release, open release, and transverse carpal ligament reconstruction were performed with tendon excursion measurements made in each of four wrist positions after each procedure. Fingertip to palm distance was also measured. The measurements of flexor tendon excursion in neutral wrist position with intact transverse carpal ligament served as the norm for each finger and as the denominator in the ratio of postoperative to preoperative excursion distances. The study confirmed the importance of the transverse carpal ligament as a flexor pulley; transection of the ligament increased the amount of flexor tendon excursion necessary to achieve finger flexion and fingertip-to-palm contact. Tendon excursion/digital flexion improved after transposition flap repair. Neither open nor endoscopic carpal tunnel release conferred any particular benefit to flexor tendon excursion postoperatively. The proximal palmar aponeurosis does not seem to have the same pulley effect as the transverse fibers of the distal palm. PMID- 9727444 TI - Accompanying arteries of the cutaneous veins and cutaneous nerves in the extremities: anatomical study and a concept of the venoadipofascial and/or neuroadipofascial pedicled fasciocutaneous flap. AB - The arterial anatomy of the accompanying arteries of the cutaneous veins and cutaneous nerves in the extremities was investigated in 10 fresh cadavers that had been injected with a lead oxide-gelatin mixture throughout the entire body. It is well known that cutaneous nerves have neurocutaneous perforators, but it was found that cutaneous veins also have their own accompanying arteries as well. The accompanying arteries of the cutaneous veins had branches not only to the vein wall, but also to the skin, i.e., venocutaneous perforators. Based on these findings, the concept of the adipofascial pedicled fasciocutaneous flap using the accompanying arteries of the cutaneous veins, cutaneous nerves, or both was proposed. PMID- 9727445 TI - Autoclaved bone for craniofacial reconstruction: effects of supplementation with bone marrow or recombinant human fibroblast growth factor-2. AB - Replantation of resected bone after autoclaving is employed by many units in both craniofacial and orthopedic tumor reconstructive surgery. The procedure is attractive because it maintains the original anatomy, is reliable, and provides a good immediate clinical result. However, doubts have been raised about the ability of the autoclaved bone to revitalize. The present study aimed to explore, in an animal model, the revitalization of autoclaved bone and to determine whether it would be possible to enhance graft revitalization using either autogeneic bone marrow or recombinant human fibroblast growth factor-2, a peptide with stimulatory effects on both endothelial and osteogenic cells. Twenty-eight adult rats were subjected to bilateral parietal cranioplasties (4 x 6 mm), and 75 percent of the grafts were subjected to autoclaving (121 degrees C; 20 minutes) and subsequently treated randomly according to one of the following strategies: no further treatment, or supplementation with bone marrow or recombinant human fibroblast growth factor-2. The remaining grafts were replanted as fresh autografts. The results were evaluated after 4 and 12 weeks by radiologic, histologic, and histomorphometric analyses. After 4 weeks, no major differences were observed between treatments. At 12 weeks, however, no distinction in graft revitalization between autografts and autoclaved grafts supplemented with recombinant human fibroblast growth factor-2 was observed, whereas autoclaved grafts with or without bone marrow displayed significantly less revitalization compared with autografts. The results indicate that autoclaved bone will revitalize by remodeling, and that the efficacy of this process can be increased significantly by simultaneous supplementation with recombinant human fibroblast growth factor-2. PMID- 9727446 TI - Nondisruptive, in vivo method for biomechanical characterization of linear incision wound healing: preliminary report. AB - Previous work on in vivo biomechanical characterization of soft tissues and wound healing has led to the development of a methodology for nondisruptive, in vivo biomechanical analysis of linear incision wounds. The purpose of this preliminary study was to define nondisruptive biomechanical parameters that characterize progressive healing and compare them with an in vivo disruptive parameter of ultimate pressure at failure (in vivo strength). Male Sprague-Dawley rats (n = 60), each weighing 250 to 300 gm, were anesthetized and underwent creation of paired full-thickness linear incision wounds. The rats were divided into three groups (n = 10/group per time period): control group, nothing applied to either wound; carrier group, bovine serum albumin applied to each wound; transforming growth factor-beta2 group, transforming growth factor-beta2 in bovine serum albumin applied to each wound. On postoperative days 5 and 10, rats from each group underwent in vivo biomechanical testing using the Dimensional Analysis System (Dimensional Analysis Systems, Inc., Nashville, Tenn.). This computer driven device integrates a video camera and processor with a vacuum controller, valve, and transducer to provide measurements of tissue deformation (in millimeters) and negative pressure (mmHg) as a multiaxial stress (vacuum) is applied to a wound. On each rat, one incision was tested disruptively and the other incision was tested nondisruptively. Disruptive data were measured as ultimate pressure (mmHg) at failure, or the amount of pressure required to disrupt the wound. Nondisruptive data were measured for tissue stiffness (kiloPascals) during application of negative pressure (maximum, 80 mmHg). On postoperative day 5, wounds treated with transforming growth factor-beta2 had significantly increased in vivo wound strength compared with carrier wounds. The nondisruptive parameter of tissue stiffness was also significantly increased for the transforming growth factor-beta2 treated wounds, thus supporting the disruptive data. On postoperative day 10, there was no difference in mean wound strength or mean tissue stiffness among any of the groups. These preliminary data represent the first report of in vivo, nondisruptive biomechanical characterization of linear incision wounds. The results suggest that through in vivo measurements of tissue stiffness, differences can be detected between treatment groups. Because the healing wound may be characterized without the need for disruption, this methodology should allow for consecutive, in vivo biomechanical testing of wounds in future wound healing studies. PMID- 9727447 TI - Determinations of strength of synthetic hydroxyapatite ceramic implants. AB - To study the physical strengths of various types of synthetic high porosity hydroxyapatite plates, we constructed samples of a fixed shape, 8 X 25 mm in size, with varying degrees of thickness (3 to 10 mm) and porosity (40, 50, and 60 percent), as well as samples with varying degrees of curvature and samples constructed with computer-aided design-computer-aided manufacturing (CAD-CAM) technology based on a real-size model made with laser lithography from computed tomography data. The strength of the samples was studied with the three-point bending test and crush tests. Studies showed that strength decreases with increasing porosity and increases with increasing thickness. In addition, results of testing plates of varying shapes and degrees of curvature revealed that the effects of these variations were small and that when the width and thickness were held constant, changing the curvature of the entire unit (from a height of 20 to 30 mm) or altering both sides had no remarkable effect on strength. On the other hand, strength testing of plates of various shapes and thicknesses constructed from clinical computed tomography data revealed that minimum optimization of the parameters was achieved when plates had a porosity of about 40 percent and a thickness of about 8 mm. PMID- 9727448 TI - Necrotizing fasciitis of the face and neck. AB - Necrotizing fasciitis is a rare infection in the surgical and infectious medical services, but when it is present, in most cases, the affected areas are the lower extremities and the abdominal and thoracic walls, and more frequently in perineum and genital areas, which is commonly known as Fournier's gangrene. This type of infection is extremely rare in the face and neck, because the great vascularity in these regions makes the tissues rarely susceptible to ischemia and infection, even when large flaps are lifted in reconstructive and aesthetic procedures. We present a case of necrotizing fasciitis in face and neck, with emphasis on the medical and surgical treatment that should be based on antibiotic therapy of broad spectrum; secondary reconstructive procedures should be planned. PMID- 9727449 TI - Free flap repair of septal perforation. PMID- 9727450 TI - Hyperbaric oxygen therapy for cutaneous/soft-tissue zygomycosis complicating diabetes mellitus. AB - A 24-year-old female diabetic patient was hospitalized because of ketoacidosis and a necrotic wound on the hand. Debridement and antibiotic therapy failed to halt the process. After demonstration of Mucor in cultures from the wound, the patient underwent extensive surgery and amphotericin B was administered. When the necrotic process continued despite these measures, adjunctive hyperbaric oxygen (100% O2 at 2.5 ATA for 90 minutes) was administered daily for a total of 21 treatment sessions. She gradually improved, and at 2 months follow-up most of the wound had healed. Although the mortality rate of cutaneous/soft-tissue zygomycosis is markedly lower than that of the rhinocerebral form, morbidity is still considerably high. Successful use of hyperbaric oxygen has been reported in rhinocerebral zygomycosis, and it may have been of benefit in this high-risk patient by preventing local and systemic spreading of the fungus. This report is the first case of the use of hyperbaric oxygen for cutaneous/soft-tissue zygomycosis. It is suggested that hyperbaric oxygen be considered for this indication in diabetic patients as an adjunct to surgery and amphotericin B. PMID- 9727451 TI - The use of cultured autologous keratinocytes with integra in the resurfacing of acute burns. PMID- 9727452 TI - Butterfly-winged island temporoparietal fascial flap for secondary auricular reconstruction. PMID- 9727453 TI - The extended submental island lip flap: an alternative for esophageal repair. AB - An esophagocutaneous fistula after total laryngectomy in a radiated field is rare. A 62-year-old man, with a history of T2N0 M0 laryngeal carcinoma, was treated with radiation therapy. He subsequently developed recurrent disease and underwent total laryngectomy. A complication of his total laryngectomy was a high esophagocutaneous fistula. The patient had no evidence of other disease. A functional repair was achieved by extending the submandibular arterial flap to incorporate the central third of the lower lip as a mucosomyocutaneous flap. This extension of the submandibular artery flap may preclude the need for jejunal free tissue transfer in some patients with esophagocutaneous fistula. PMID- 9727455 TI - Subperiosteal face lift: a 200-case, 4-year review. AB - The subperiosteal face lift is a procedure designed to rejuvenate the upper and middle thirds of the face. Herein is reviewed a 4-year series of 200 consecutive patients who have undergone a subperiosteal face lift with a special emphasis on handling of the zygomatic arch. The main operative indication was significant ptosis of the midface soft tissue. Dissection of the maxilla, zygoma, periorbital areas, and the anterior arch was carried out through either a gingivo-buccal sulcus incision (39 cases) or a subciliary incision (161 cases). Dissection of the posterior arch was carried out in a plane superficial to the innominate fascia. A back-cut was made in the superficial musculoaponeurotic system and subcutaneous tissue down to midtragus, and a subperiosteal tunnel was entered by piercing through the posterior arch periosteum. By using a Cottle elevator (sweeping superiorly and inferiorly), the arch dissection was completed in a posterior to anterior direction. All patients underwent a concurrent brow lift (190 endoscopically and 10 by means of coronal incision). The forehead incision was used to dissect the lateral orbital rims. Twelve patients (6 percent) had undergone a previous rhytidectomy. All but four patients were women and ranged in age from 34 to 76 years (mean, 54+/-11). Mean follow-up period was 27 months (1 to 41 months). The postoperative complication rate was 5 percent and included transient frontal branch weakness (n = 2), resolved at 41 and 71 days postoperatively; hematoma (n = 2); transient infraorbital nerve paresthesia (n = 1); asymmetrical smile (n = 3); and facial tics (n = 2). Two patients (1 percent) required a secondary surgery on their brows or midface. An upper blepharoplasty was needed in 26 patients (13 percent). The overall aesthetic results were excellent, with good elevation of the eyebrows, lateral canthus, and the midface soft tissues. In conclusion, the subperiosteal face lift is a procedure designed to rejuvenate the upper and middle thirds of the face. Approaching the arch posteriorly and in a systematic fashion simplifies the procedure and reduces the risk of facial nerve injury. PMID- 9727454 TI - An innovative method of reconstruction of large skeletal chest wall defects. AB - The reconstruction of large skeletal defects of the chest wall is considered complex and difficult. A simple technique for this repair, using polymethylmethacrylate ribs that are prefabricated in a prosthodontic laboratory using heat cure polymer, is presented. These ribs are used as bone substitutes while reconstructing the chest wall defect. The prefabricated polymethylmethacrylate ribs have been used to reconstruct the large chest wall defects after resection of chest wall tumors in three patients. There was risk of severe paradoxical movement of the chest wall without the skeletal defect reconstruction. In all of the cases, there was no need for postoperative mechanical ventilation and there was no paradoxical movement. Cosmetically, it was highly satisfactory. In one patient, the stainless steel wire caused a sinus after 1 year of reconstruction, requiring removal of the wire and the rib, but it did not compromise the stability of the chest wall. We conclude that reconstruction of large chest wall defects with prefabricated polymethylmethacrylate ribs is simple, cost-effective, and easy to plan and execute. PMID- 9727456 TI - Nasal osteotomy and airway changes. AB - The purpose of this prospective investigation was to evaluate the factors involved in the constricting effect of nasal bone osteotomy on the nasal airway. Immediately before the osteotomy, observations were made and recorded in regard to both the position of the inferior turbinates and the length of the nasal bones. During the osteotomy, the nasal bone movement was graded and the type of osteotomy was documented. The two types of osteotomy were defined as either high to-low or low-to-low. Each side of the nose was assessed independently. Forty eight consecutive patients, 8 men and 40 women, were included in this study, providing 96 nasal sides for evaluation. There were 42 normal, 32 short, and 22 long nasal bones. The patients with short nasal bones exhibited less diminution in the airway than those patients with normal nasal bones (p < 0.05). The position of the inferior turbinates was designated as anterior in 48 sites, 12 were considered normal, and 28 were deemed posterior. The narrowing of the airway was significant when the inferior turbinates were positioned anteriorly when compared with posteriorly positioned inferior turbinates (p < 0.05). Twenty-four nasal bones were shifted slightly, 48 intermediately, and 22 significantly. Major nasal airway constriction was observed when the medial positioning of the nasal bone was significant (p < 0.05). Eighty-four osteotomies were classified as low to-low, and 12 were high-to-low. High-to-low osteotomies resulted in the least narrowing of the nasal passage (p < 0.005). It is concluded from this study that the nasal osteotomy does constrict the nasal airway in most incidences. The length of the nasal bones, the degree of nasal bone repositioning, the position of the inferior turbinates, and the type of osteotomy are definite factors contributing to airway narrowing after nasal bone osteotomy. PMID- 9727457 TI - The infradome graft: a new technique to improve dome reshaping in rhinoplasty. AB - As the open approach rhinoplasty has gained popularity, newer techniques for aggressive reshaping of the alar cartilage have become available. The most effective way to increase alar dome definition is the dome plication technique described by Daniel, which places a horizontal mattress suture, applied with incremental tension, between the medial and lateral crura. The disadvantage of this technique is that it can produce either overtightening or distortion of the cartilage, with excessive pinching of the nasal tip. Also, as it relies on a single nylon suture to fight against the cartilage's shape memory, it has the potential for relapse. The present article describes a new technique to overcome these problems. It consists of placing two parallel strips prepared from the resected alar cartilage excess under the domes. These grafted cartilages block the plication suture, thus avoiding the risk of excessive pinching of the domes and ensuring symmetry and stability. Using this technique, I have consistently obtained a nasal tip that is well defined over the nasal dorsum line in my patients. PMID- 9727458 TI - Skin tightening effects of the ultrapulse CO2 laser. AB - This study analyzed the skin tightening or contracture effect of the Ultrapulse carbon dioxide (CO2) laser on the skin of hairless guinea pigs by light and electron microscopic, histologic, and tensiometric evaluations. Two 2 X 2 cm squares of back skin were precision tattooed on each of the animals in the study (n = 12). One square served as the control, and the other square was used as experimental skin. The experimental skin was treated with three passes of the CO2 laser at 500 mJ and 5 W using a 3-mm collimated hand-piece. Skin specimens from three animals were analyzed at 1, 4, 8, and 12 weeks. After three passes, the length of the square was reduced by 27 percent, and the width was reduced by 40 percent. Over the next 12 weeks, as the animals grew, the dimensions of the control areas also increased. The laser-treated areas continued to maintain their contracted dimensions, however. By the 12th week, the laser-treated areas were 28.35 percent shorter in length and 15.5 percent shorter in width than the control areas. Histologic examination demonstrated a significantly higher content of collagen in the reticular layer, which was more compact than that of the normal skin. Electron microscopy revealed that the laser had induced shortening of the collagen fibers (7.45 percent; p = 0.026), which persisted beyond the 12th week. Laser treatment did not significantly alter the tensile strength of the skin, although, at the 8th week, the treated areas had a slightly higher tensile strength. PMID- 9727459 TI - SMAS rhytidectomy versus deep plane rhytidectomy: an objective comparison. AB - Although there are a multitude of techniques currently used for performing face lifts, there is no general agreement as to which, if any, of these techniques is most effective. There may never be a definitive answer to this issue because of the highly subjective nature of aesthetics, variability among surgeons, differences in patient anatomy, and specific patient desires. In an attempt to evaluate face lift techniques objectively, this study compares the rate of patients undergoing a tuck procedure after traditional SMAS (superficial musculoaponeurotic system) rhytidectomy to that of patients after deep plane rhytidectomy. A retrospective chart review was performed on all patients who underwent a tuck procedure following a face lift by the senior author (Kamer) between July of 1990 and January of 1997. There were 634 patients who electively underwent either a SMAS or deep plane type of rhytidectomy during the 6.5-year period; 48 patients subsequently underwent tuck operations, and adequate information was available on 44 patients. Of these, 43 were women and the average age was 57 years. The overall tuck rate from July of 1990 to January of 1997 was 7.5 percent. The tuck rate following SMAS rhytidectomy was 11.4 percent, and that following deep plane rhytidectomy was 3.3 percent. Therefore, a tuck was required 71 percent less frequently after a deep plane lift than after a SMAS lift. This was found to be a statistically significant difference with a p value of .0001 (Fisher's exact test, 2-tail). If the assumption is made that the need for a tuck procedure implies a less than optimal face lift, then the data of this study suggest that the deep plane technique is more effective than the SMAS technique. PMID- 9727460 TI - Capsulectomy. PMID- 9727461 TI - Guidelines and indications for breast implant capsulectomy. AB - This article discusses indications for performing a capsulectomy in conjunction with explantation of breast implants. This issue has rarely been addressed in the literature, and there is no consensus on guidelines to assist surgeons in deciding whether a capsulectomy is warranted. The many factors that must be weighed when considering performance of a capsulectomy are outlined, and recommendations for the explantation contexts in which capsulectomy may be considered optional or should usually be performed are given. Capsulectomy may be indicated in the majority of instances when breast implants are removed or exchanged, but the potential risks of capsule removal must always be balanced against the potential benefits. PMID- 9727462 TI - The plastic surgery research fellow: revitalizing an important asset. PMID- 9727463 TI - On the importance of being honest. PMID- 9727464 TI - The changing health care marketplace: current industry trends, new provider organizational structures, and effects on plastic surgeons. AB - Current market forces are driving the health care industry in new directions. The managed care industry is currently undergoing a market shakeout, as manifested by consolidation, increased competition, and lower profits. Medicare is fighting to remain solvent by lowering fees paid to providers, driving patients into managed care plans, and cracking down on billing irregularities. For providers, the combined effect of these trends is lower fees, increased risk-sharing, and increased overhead. Plastic surgeons face new demands in this environment. They must increase their efficiency and form new alliances with other providers. These alliances allow plastic surgeons to maintain a steady stream of patients, to manage risk, to negotiate more lucrative contracts with managed care organizations, and to increase efficiency. To achieve these alliances, plastic surgeons must alter the organizational structure of their practices. Several corporate practice models are becoming more prevalent; these include large group practices, physician practice management companies, and integrated delivery systems. Each structure has advantages for plastic surgeons, but each also requires plastic surgeons to trade varying degrees of financial and professional autonomy for market strength. PMID- 9727465 TI - Orthodox Jewish law (Halachah) and plastic surgery. PMID- 9727466 TI - The use of epinephrine in infiltrative local anesthesia for eyelid reconstruction. PMID- 9727467 TI - Senile enophthalmia and herniated lower eyelid fat. PMID- 9727468 TI - Botox use in prevention of dry eyes. PMID- 9727469 TI - Orbital septum revisited. PMID- 9727470 TI - Closed correction of convexity of the lateral crura. PMID- 9727471 TI - Palatal fractures. PMID- 9727473 TI - Phantom breast pain. PMID- 9727472 TI - Correction of tuberous breasts. PMID- 9727474 TI - Epidemiology and causation: the breast implant controversy. PMID- 9727475 TI - The rectus abdominis musculoperitoneal and posterior rectus sheath peritoneal flaps. PMID- 9727476 TI - Hair growth in the vagina after reconstruction with pudendal thigh flaps in congenital vaginal agenesis. PMID- 9727477 TI - Ultrasound-assisted lipoplasty or internal ultrasonic lipotripsy? PMID- 9727478 TI - Epithelioid sarcoma: a confusing rare tumor on the hand. PMID- 9727479 TI - Experiments using animals. PMID- 9727480 TI - The ten commandments of office-based surgery with anesthesia. PMID- 9727481 TI - Anesthesia risks in patients who have had antiobesity medication. PMID- 9727482 TI - VDJ recombination: a transposase goes to work. PMID- 9727483 TI - Somatic hypermutation, transcription, and DNA mismatch repair. PMID- 9727484 TI - Mesoderm induction: a postmodern view. PMID- 9727485 TI - Roles for proteolysis and trafficking in notch maturation and signal transduction. PMID- 9727486 TI - Structure of the histone acetyltransferase Hat1: a paradigm for the GCN5-related N-acetyltransferase superfamily. AB - We have solved the crystal structure of the yeast histone acetyltransferase Hat1 acetyl coenzyme A (AcCoA) complex at 2.3 A resolution. Hat1 has an elongated, curved structure, and the AcCoA molecule is bound in a cleft on the concave surface of the protein, marking the active site of the enzyme. A channel of variable width and depth that runs across the protein is probably the binding site for the histone substrate. A model for histone H4 binding by Hat1 is discussed in terms of possible sources of specific lysine recognition by the enzyme. The structure of Hat1 provides a model for the structures of the catalytic domains of a protein superfamily that includes other histone acetyltransferases such as Gcn5 and CBP. PMID- 9727487 TI - Crystal structure of a GCN5-related N-acetyltransferase: Serratia marcescens aminoglycoside 3-N-acetyltransferase. AB - The X-ray structure of a canonical GCN5-related N-acetyltransferase (GNAT), Serratia marcescens aminoglycoside 3-N-acetyltransferase, bound to coenzyme A (CoA) has been determined at 2.3 A resolution. The single domain alpha/beta protein resembles a cupped right hand wrapped around a cylinder and consists of a highly curved, six-stranded beta sheet of mixed polarity that is sandwiched between four alpha helices. The structure includes all four conserved GNAT motifs (C, D, A, and B) and represents the catalytic core of this large enzyme superfamily. Acetyl CoA recognition is mediated by a betaalpha structure derived from GNAT motif A, which presents an invariant Arg/Gln-X-X-Gly-X-Gly/Ala segment for hydrogen bonding with the cofactor. Motif B contributes acidic residues to the binding site for the positively charged antibiotic substrate. PMID- 9727488 TI - Retrohoming of a bacterial group II intron: mobility via complete reverse splicing, independent of homologous DNA recombination. AB - The mobile group II intron of Lactococcus lactis, Ll.LtrB, provides the opportunity to analyze the homing pathway in genetically tractable bacterial systems. Here, we show that Ll.LtrB mobility occurs by an RNA-based retrohoming mechanism in both Escherichia coli and L. lactis. Surprisingly, retrohoming occurs efficiently in the absence of RecA function, with a relaxed requirement for flanking exon homology and without coconversion of exon markers. These results lead to a model for bacterial retrohoming in which the intron integrates into recipient DNA by complete reverse splicing and serves as the template for cDNA synthesis. The retrohoming reaction is completed in unprecedented fashion by a DNA repair event that is independent of homologous recombination between the alleles. Thus, Ll.LtrB has many features of retrotransposons, with practical and evolutionary implications. PMID- 9727489 TI - DNA transposition by the RAG1 and RAG2 proteins: a possible source of oncogenic translocations. AB - The RAG1 and RAG2 proteins are known to initiate V(D)J recombination by making a double-strand break between the recombination signal sequence (RSS) and the neighboring coding DNA. We show that these proteins can also drive the coupled insertion of cleaved recombination signals into new DNA sites in a transpositional reaction. This RAG-mediated DNA transfer provides strong evidence for the evolution of the V(D)J recombination system from an ancient mobile DNA element and suggests that repeated transposition may have promoted the expansion of the antigen receptor loci. The inappropriate diversion of V(D)J rearrangement to a transpositional pathway may also help to explain certain types of DNA translocation associated with lymphatic tumors. PMID- 9727490 TI - Repression of heat shock transcription factor HSF1 activation by HSP90 (HSP90 complex) that forms a stress-sensitive complex with HSF1. AB - Heat shock and other proteotoxic stresses cause accumulation of nonnative proteins that trigger activation of heat shock protein (Hsp) genes. A chaperone/Hsp functioning as repressor of heat shock transcription factor (HSF) could make activation of hsp genes dependent on protein unfolding. In a novel in vitro system, in which human HSF1 can be activated by nonnative protein, heat, and geldanamycin, addition of Hsp90 inhibits activation. Reduction of the level of Hsp90 but not of Hsp/c70, Hop, Hip, p23, CyP40, or Hsp40 dramatically activates HSF1. In vivo, geldanamycin activates HSF1 under conditions in which it is an Hsp90-specific reagent. Hsp90-containing HSF1 complex is present in the unstressed cell and dissociates during stress. We conclude that Hsp90, by itself and/or associated with multichaperone complexes, is a major repressor of HSF1. PMID- 9727491 TI - Bid, a Bcl2 interacting protein, mediates cytochrome c release from mitochondria in response to activation of cell surface death receptors. AB - We report here the purification of a cytosolic protein that induces cytochrome c release from mitochondria in response to caspase-8, the apical caspase activated by cell surface death receptors such as Fas and TNF. Peptide mass fingerprinting identified this protein as Bid, a BH3 domain-containing protein known to interact with both Bcl2 and Bax. Caspase-8 cleaves Bid, and the COOH-terminal part translocates to mitochondria where it triggers cytochrome c release. Immunodepletion of Bid from cell extracts eliminated the cytochrome c releasing activity. The cytochrome c releasing activity of Bid was antagonized by Bcl2. A mutation at the BH3 domain diminished its cytochrome c releasing activity. Bid, therefore, relays an apoptotic signal from the cell surface to mitochondria. PMID- 9727493 TI - Motor neuron-derived retinoid signaling specifies the subtype identity of spinal motor neurons. AB - The diversification of neuronal cell types in the vertebrate central nervous system depends on inductive signals provided by local organizing cell groups of both neural and nonneural origin. The influence of signals provided by postmitotic neurons on the fate of neurons born at subsequent development stages, however, remains unclear. We provide evidence that a retinoid-mediated signal provided by one subset of early-born spinal motor neurons imposes a local variation in the number of motor neurons generated at different axial levels and also specifies the identity of a later-born subset of motor neurons. Thus, in the vertebrate central nervous system the distinct fates of late-born neurons may be acquired in response to signals provided by early-born neurons. PMID- 9727492 TI - Cleavage of BID by caspase 8 mediates the mitochondrial damage in the Fas pathway of apoptosis. AB - We report here that BID, a BH3 domain-containing proapoptotic Bcl2 family member, is a specific proximal substrate of Casp8 in the Fas apoptotic signaling pathway. While full-length BID is localized in cytosol, truncated BID (tBID) translocates to mitochondria and thus transduces apoptotic signals from cytoplasmic membrane to mitochondria. tBID induces first the clustering of mitochondria around the nuclei and release of cytochrome c independent of caspase activity, and then the loss of mitochondrial membrane potential, cell shrinkage, and nuclear condensation in a caspase-dependent fashion. Coexpression of BclxL inhibits all the apoptotic changes induced by tBID. Our results indicate that BID is a mediator of mitochondrial damage induced by Casp8. PMID- 9727494 TI - The role of maternal VegT in establishing the primary germ layers in Xenopus embryos. AB - VegT is a T-box transcription factor whose mRNA is synthesized during oogenesis and localized in the vegetal hemisphere of the egg and early embryo. We show that maternally expressed VegT controls the pattern of primary germ layer specification in Xenopus embryos. Reduction of the maternal store completely alters the fates of different regions of the blastula so that animal cell fate is changed from epidermis and nervous system to epidermis only, equatorial cell fate is changed from mesoderm to ectoderm, and vegetal cell fate is changed from endoderm to mesoderm and ectoderm. Vegetal cells lose their capacity both to form endoderm and to release mesoderm-inducing signals. These results show that a single maternally expressed gene controls the patterning of the Xenopus blastula. PMID- 9727495 TI - Crystal structure of the hexamerization domain of N-ethylmaleimide-sensitive fusion protein. AB - N-ethylmaleimide-sensitive fusion protein (NSF) is a cytosolic ATPase required for many intracellular vesicle fusion reactions. NSF consists of an amino terminal region that interacts with other components of the vesicle trafficking machinery, followed by two homologous ATP-binding cassettes, designated D1 and D2, that possess essential ATPase and hexamerization activities, respectively. The crystal structure of D2 bound to Mg2+-AMPPNP has been determined at 1.75 A resolution. The structure consists of a nucleotide-binding and a helical domain, and it is unexpectedly similar to the first two domains of the clamp-loading subunit delta' of E. coli DNA polymerase III. The structure suggests several regions responsible for coupling of ATP hydrolysis to structural changes in full length NSF. PMID- 9727496 TI - Relocation of the t-SNARE SNAP-23 from lamellipodia-like cell surface projections regulates compound exocytosis in mast cells. AB - For regulated secretion, mast cells and several other cell types utilize compound exocytosis, a combination of granule-plasma membrane and granule-granule fusions. The molecular machinery that controls this massive export process has not been identified. We report that SNAP-23, a t-SNARE related to SNAP-25, relocates in response to stimulation from plasma membrane lamellipodia-like projections to granule membranes in permeabilized mast cells. While relocation is a prerequisite for secretion, it can occur without membrane fusion and will expedite a subsequent secretory response. After relocation, SNAP-23 is required for exocytosis, implying a crucial role in promoting membrane fusion. Thus, relocation of this SNARE regulates compound exocytosis and links granule-plasma membrane and granule-granule fusions. PMID- 9727497 TI - Coupling cofactor--a novel regulatory component in G protein-mediated signalling? AB - The model for studying the mechanism of G protein-mediated signalling cannot account for the observation that high-affinity binding of agonists to many different receptors is not dissociated by the addition of high concentrations of guanine nucleotides. Using the cerebral A1-adenosine receptor as a model system, we have recently identified a component which is responsible for this phenomenon. This protein, termed the coupling cofactor, can be solubilized from brain membranes and chromatographically resolved from both the G proteins and the receptor. Following reconstitution into appropriate acceptor membranes, the coupling cofactor confers resistance of high-affinity agonist binding to guanine nucleotides. The coupling cofactor acts as a brake and limits receptor-dependent signal amplification; in addition, it is a candidate for participating in the higher level organization of receptors and G proteins in membranes and in the membrane-delimited cross-talk between individual receptors. Here, we present a working hypothesis on the possible biological roles of the coupling cofactor. PMID- 9727498 TI - Critical remarks on the international WHO classification of rodent central nervous system (CNS) tumours. PMID- 9727500 TI - Monitoring of the boron neutron-capture reaction by a simple animal model. AB - Sodium borocaptate (BSH, Na2Bl2HllSH), a slow neutron-capture compound, was injected into the left forebrain ventricle of 1-week-old rats (150 microg BSH/3 microl phosphate buffered saline). After 90 min, the animals were irradiated by epithermal neutrons (LVR-15 nuclear reactor in Rez near Prague, flux density 8.8 x 10(7) neutrons cm-2 s-1, 8 MW reactor power, 8.2 cGy/min) for 5, 10 or 20 min. The brains were examined histologically 8 h after irradiation. In animals irradiated for 5 to 10 min (41 and 82 cGy-Eq, respectively) lethal damage of cells was found in the external granular layer of the cerebellum and the subependymal layer of the forebrain. Irradiation for 20 min (164 cGy-Eq) caused more extensive destruction of cell populations in these regions and, in addition, dead cells appeared also in the more differentiated postmitotic compartments, namely the deeper layers of the cerebellum, layers II/III of the cerebral cortex and corpus callosum. In the forebrain periventricular layer, the extent of cell damage was declining towards the olfactory bulbs. In intact animals, as well as in those injected only with the 150 microl phosphate buffered saline, the radiation damage was low and limited only to the most sensitive dividing populations of the cerebellum and the forebrain. The study demonstrates a differentiation-dependent damage of the rat brain cells by alpha particles and presents a simple model for evaluation of the biological effectiveness of slow neutron beams constructed for neutron-capture therapy of tumors. PMID- 9727499 TI - Neutron-capture therapy of brain tumours: neutron sources, neutron-capture drugs, biological tests and clinical perspectives in the Czech Republic. AB - The paper reviews neutron sources, chemical compounds and clinical perspectives of the boron neutron-capture therapy of brain tumours. Special attention is paid to the physical characteristics and biological effectiveness of the epithermal neutron beam constructed at the LVR-15 nuclear reactor at Rez near Prague. PMID- 9727501 TI - The effect of drugs on the mortality of mice inoculated with Friend leukaemia virus or toxoplasma gondii. AB - Infection and tumors provoke substantial changes accompanied with the disbalance of many neuroendocrine factors which in their summarizing effects influence the life span of animals. Our previous results showed enhanced mortality after one injection of morphine in association with Friend leukaemia virus infection. The aim of this study was to examine the effects of some other opioids (pethidine and pentazocine) and an acetylcholine esterase inhibitor neostigmine on the survival of animals under two conditions: (1) Friend leukaemia virus infection which mostly depressed immune functions, and (2) Toxoplasma gondii infection which in general enhanced the immune status. In contrast to our previous observation with morphine, the mortality induced by single doses of pethidine (150 mg/kg) or pentazocine (50-75 mg/kg) was unchanged during the Friend leukaemia virus infection. A single injection of neostigmine (0.42 or 0.56 mg/kg) was significantly more lethal in DBA-2 mice infected with Friend leukaemia virus. Neostigmine in doses of 0.33 and 0.4 mg/kg caused death in 46 % and 57 %, respectively, of animals infected with Toxoplasma gondii which was significantly higher in comparison with only 8 % and 12.5 % in control groups. Pethidine (150 mg/kg) killed 70 % of Toxoplasma gondii infected animals and even 90 % of non infected mice. Thus, the Friend leukaemia virus and Toxoplasma gondii infections increased toxicity only of some drugs which may, at least partly, be associated with altered immune status during infection and involvement of the cholinergic system. PMID- 9727502 TI - The effect of dopaminergic agents on cell-mediated immune response in mice. AB - The aim of the study was to determine the effect of selective dopaminergic agents [(+/-)-SKF-81297 (D1 agonist), R(-)-2,10,11-trihydroxy-N-propyl-noraporphine (D2 agonist), pergolid (D agonist), R(+)-SCH-23390 (D1 antagonist), S(-)-eticlopride (D2 antagonist) and cis-(Z)-flupenthixol (D antagonist)] on cell-mediated immune response in vivo and in vitro and to verify the presence of dopamine receptors on murine splenocytes. The tested dopaminergic compounds exhibited a pronounced inhibitory effect on T-dependent immunity. They suppressed alloantigen-induced immune response in vivo and in vitro, IL-2 production was also markedly reduced. No substantial difference was found between the effect of dopamine agonists and antagonists or among ligands of subtypes of dopamine receptors. The effect of dopaminergic agents in vitro indicates a direct interaction with immunocompetent cells at the peripheral level. As the binding studies did not confirm the presence of dopamine receptors on splenocytes, the immunosuppressive efficacy of dopaminergic agents does not seem to be mediated via specific dopamine receptors. PMID- 9727503 TI - Neuroimmunomodulation of natural killer (NK) cells by ergot alkaloid derivatives. AB - Ergot alkaloids (EAs), products of Claviceps spp., are widely used in various fields of clinical medicine (neurology, psychiatry, endocrinology). In the present work we studied the neuroimmunomodulative effect of EAs on activation of NK cells and their signalling pathways. Furthermore, the killing capability of rat NK cells in vitro was examined in the presence of glycosidic derivatives of elymoclavine, agroclavine, and liposome-encapsulated EAs. The engagement of appropriate NK cell membrane receptors by EAs cause an indirect enhancement of adenylyl cyclase system through inhibition of G-protein al,2-subunit (up to 50 % of control values). All of the tested EAs enhanced the rat NK cell-mediated cytotoxic activity in vitro, particularly against target cells of astrocyte origin (C-6 glioma). The present results argue for a possible EA immunomodulatory role of cell-mediated immunity in tumour regression processes. PMID- 9727504 TI - In vitro early allogeneic reaction of murine brain cortex cells. AB - An allogeneic reaction among brain cortex cells (mixed reaction) was demonstrated previously by H-2 alloantigen-induced uncoupling of oxidative metabolism (Kovaru Med. Biol. 58: 273, 1980). In the present study we have demonstrated that alloantigen already increased cell surface Na+,K+-ATPase activity after 100 min when the enzyme activation was highest at Mg2+/ATP ratio 4: 1. The allogeneic cell reaction was accompanied by an elevation of membrane lipid fluidity and probably also by a thermotropic lipid phase transition which might influence the membrane lipid-dependent Na+,K+-ATPase activity, while Mg2+-ATPase remained unaffected. Furthermore, the effects of proteins and peptides released into the supernatant during the allogeneic reaction were analyzed in brain cortex cells. One of the isolated active peptide fractions, FA (m.w. lower than 2.5 kD), was able to enhance Na+,K+-ATPase activity as well as to block K+-evoked O2 uptake by brain cortex cells. Thus the FA fraction simulated primary allorecognition events. The data indicate that various brain cell surface domains were influenced by a regulatory peptide fraction of the cytokine type during the early phase of allogeneic reaction. Allorecognition among brain cortex cells is directed against functionally important metabolic reactions. PMID- 9727505 TI - Cell surface enzymes of brain cortex cells or lymphocytes during early allogeneic reaction. AB - The aim was to study the role of major histocompatibility complex (MHC), in mice named H-2, during early allogeneic reactions (AR) of brain cortex cells or lymphocytes. We used neuronal and glial enriched perikarya, spleen and thymus lymphocytes or their subpopulations. Rat AR was also assayed between C-6 astrocytoma cells and spleen lymphocytes. We demonstrated that: 1) H-2 dependent stimulation of Na+,K+-ATPase and ouabain-sensitive K+-dependent p nitrophenylphosphatase (K+-pNPPase) activities represented specific response in both AR of unseparated brain cells or lymphocytes. On the other hand, non specific AR-induced stimulation of Ca2+-ATPase activity was observed. 2) Allogeneic enriched glial fractions reacted similarly by the same enzyme activation in contrast to no change in AR between enriched neuronal fractions. Allorecognition ability of glial cells was confirmed by AR between C-6 astrocytoma cells and lymphocytes. 3) Mature thymus lymphocytes exerted alloreactivity by specific activation of Na+,K+-ATPase or K+-pNPPase, in contrast to no change in AR between immature lymphocyte subpopulations. 4) MHC Class II monoclonal antibody inhibited Na+,K+-ATPase and K+-pNPPase activities in brain cells as well as in thymus and spleen lymphocytes in a dose-dependent manner. Results support former studies about alloantigen-induced uncoupling in brain oxidative cortex metabolism (Kovaru Med. Biol. 58: 273, 1980) via Na+,K+-ATPase and K+-pNPPase inhibition by mechanism which can mimic MHC restriction. PMID- 9727506 TI - Peptide cytokines in CNS and the immune system. AB - This study describes the effects of cytokine peptides released into the supernatant during an early allogeneic reaction (AR) of mouse spleen lymphocytes or brain cortex cells which differ in their major histocompatibility complex (MHC). The peptides were isolated by ultrafiltration, liquid chromatography and HPLC. We found that both peptides stimulated the cell surface Na+,K+-ATPase and Ca2+-ATPase activities of quiescent spleen lymphocytes in vitro and mimicked early allogeneic cell interactions. Both brain and spleen AR peptides inhibited Concanavalin A-stimulated spleen lymphocyte proliferation, whereas 3H-TdR incorporation into DNA of the E7 neuroblastoma cell line was stimulated by these peptides. The peptide isolated from the supernatant of the allogeneic brain cell reaction inhibited phagocytosis in phorbol myristate-stimulated LA5-9/8 mouse macrophage cell line. Immunosuppressive activity of spleen AR peptide is supported by inhibition of spontaneous E rosette formation by lymphocytes. The immunosuppressive effect of isolated peptide cytokines on lectin-activated lymphocytes was comparable with the serum thymic factor (FTS, Lenfant et al. 1983). These changes demonstrate the pleiotropic cytokine actions mediated by plasma membrane of immune system and brain cells. PMID- 9727507 TI - Apoptotic damage of DNA in human leukaemic HL-60 cells treated with C2-ceramide was detected after G1 blockade of the cell cycle. AB - Apoptosis was induced by treatment of HL-60 cells with C2-ceramide. Apoptotic damage of DNA was detected according to the sub-G1 peak on a flow cytometer, according to the typical morphology and according to the DNA fragmentation "ladder" after gel electrophoresis. It was shown that the apoptotic cleavage followed after G1 blockade of the cell cycle. A high correlation coefficient (rs=0.957) was found between the percentages of G1 blocked cells and apoptotic cells. This high correlation together with the appearance of the sub-G1 peak suggests that the G1 blocked HL-60 cells were subject to apoptotic death. It is deduced that the mechanisms leading to G1 blockade of the cell cycle and activation of apoptosis in HL-60 cells are interconnected. PMID- 9727508 TI - Estimation of apoptosis in C6 glioma cells treated with antidepressants. AB - Gel electrophoresis of DNA was used for estimation of DNA changes caused in C6 glioma cells by treatment with psychotropic drugs (imipramine, amitryptiline and fluoxetine). Some discrete bands containing a population of short DNA fragments appeared after 1 and 5 days of cultivation. Apoptotic DNA breaks were verified at single cell level using the TUNEL test in cells treated with fluoxetine. PMID- 9727509 TI - Granular corneal dystrophy with homozygous mutations in the kerato-epithelin gene. AB - PURPOSE: To report a family with several members affected with granular corneal dystrophy Groenouw type 1. Three members of the family were affected with a severe placoid type of corneal dystrophy. To determine the relationship between gene mutations and phenotypic variations of the disease, we analyzed the kerato epithelin gene. METHODS: The pedigree included a consanguineous marriage of two affected individuals. The three family members affected with a severe form of corneal dystrophy were offspring of these parents. However, the phenotype of other affected family members was typical granular corneal dystrophy. We isolated genomic DNA from leukocytes of the family members. Exons of the keratoepithelin gene were amplified by the polymerase chain reaction and were analyzed using the single-strand conformation polymorphism technique. Mutations were identified by direct sequencing method and restriction digestion analysis. RESULTS: The three severely affected family members exhibited homozygous mutations at codon 555 (arginine to tryptophan) in the keratoepithelin gene, whereas those with typical granular corneal dystrophy had the heterozygous mutation at the same codon. Unaffected family members did not have the mutation. CONCLUSIONS: We determined that the severe phenotype of granular corneal dystrophy is caused by homozygous mutations in the kerato-epithelin gene. Clinical manifestation of the severe phenotype is a placoid type of corneal dystrophy and early recurrence after surgery. Granular corneal dystrophy appears to be the first ophthalmic disease in which homozygosity for a dominant allele has been genetically identified. PMID- 9727510 TI - Intraoperative crystallization on the intraocular lens surface. AB - PURPOSE: To report a physician survey, laboratory studies, and clinical observations of intraoperative crystallization on the surface of the intraocular lens (IOL). METHOD: We sent a survey to all ophthalmologists in the states of Wyoming, Idaho, Montana, Utah, and Colorado asking whether crystallization on the IOL surface had occurred in any of their patients and what viscoelastics, IOLs, and other solutions were used. All returned surveys were tabulated and analyzed by standard statistical means. A sample of crystallization from an IOL on a glass slide submitted by a physician was analyzed to ascertain the relative elemental composition. During in vitro laboratory studies, BSS Plus (Alcon Surgical, Fort Worth, Texas) and BSS (Alcon Surgical) were measured and analyzed for precipitation. Healon GV (Pharmacia/Upjohn, Kalamazoo, Michigan) and calcium chloride were combined in various solutions and examined for precipitate formation. Silicone IOLs and plate silicone were placed in different BSS and BSS Plus solutions with different viscoelastics and varying calcium concentrations. In seven patients, prominent crystallization on an IOL surface was examined, photographed, and followed for up to 3 years. RESULTS: Two hundred six surveyed ophthalmologists returned 181 surveys (88%) and reported 29,609 cataract surgeries, with IOL implantation with 22 eyes (0.07%) (22 patients) in which intraoperative crystallization was observed on the IOL surface during 1993. The survey indicated there was a correlation with BSS Plus (chi-square = 4.9, P = .0268) and silicone IOLs (chi-square = 6.8, P = .0093). The sample showed the cation to be calcium. CONCLUSION: Crystallization on the IOL surface during cataract surgery is a rare occurrence that may be associated with calcium as the cation related to an osmotic gradient around the IOL with increased calcium concentration. If encountered surgically, the lens should be exchanged in the operating theater after irrigating the anterior chamber with BSS and completely filling the capsular bag with a low molecular weight viscoelastic. PMID- 9727511 TI - Five-year results of a randomized, prospective, clinical trial of diode vs argon laser trabeculoplasty for open-angle glaucoma. AB - PURPOSE: To evaluate the safety and efficacy of laser trabeculoplasty (LTP) with a semiconductor diode laser (810 nm, [DLT]) vs an argon blue-green laser (488 to 514 nm, [ALT]). METHODS: In a prospective, randomized clinical trial, 50 eyes of 46 patients with uncontrolled open-angle glaucoma on maximally tolerated medical therapy were treated and followed at regular intervals for 5 years. Fifty laser spots were applied over 180 degrees using either maximal laser power or sufficient power to produce blanching or a small bubble (570 to 850 mW, DLT; 400 to 1,100 mW, ALT). We performed DLT using a 100-microm spot size, a 0.5-second exposure, and a Ritch lens; we conducted ALT with a 50-microm spot, a 0.1-second exposure, and a Goldmann lens. Patients in the study were followed until trabeculectomy was required. RESULTS: The mean follow-up times +/- SD for all eyes were 38.6 +/- 5.4 months, DLT (n = 22; range, 1 to 68 months) and 35.5 +/- 4.8 months, ALT (n = 28; range, 1 to 66 months). Those in the diode laser group (n = 16) who had more than 1 year of follow-up were tracked for 49.4 months, and those in the argon laser group (n = 21) were tracked for 45.8 months. There were no significant differences in the mean pretreatment intraocular pressures (IOPs): 21.2 mm Hg, DLT (n = 22) and 21.5, ALT 21.5 mm Hg (n = 28); P = .81] or in mean final IOPs (15.7 mm Hg, DLT and 17.1 mm Hg, ALT; P = .19). Time to surgical failure showed no significant differences, with 50% of the DLT eyes and 58% of the ALT eyes surviving at 5 years (P = .59). CONCLUSION: In eyes with open-angle glaucoma and unsatisfactory IOP control on maximally tolerated medical therapy, DLT and ALT are equally effective in lowering IOP over a 5-year period. PMID- 9727513 TI - Ultrasound biomicroscopic detection of anterior ocular segment foreign body after trauma. AB - PURPOSE: To describe the role of ultrasound biomicroscopy in the detection and localization of foreign bodies in anterior ocular segment foreign body after trauma. METHODS: In a prospective study, ultrasound biomicroscopy was performed in five eyes of five consecutive patients with suspected anterior ocular segment foreign body. RESULTS: In all five eyes, ultrasound biomicroscopy detected and precisely localized small foreign bodies (metallic in two eyes, stone in one eye, plastic in one eye, and ceramic in one eye) in the cornea (one eye), superficial sclera (one eye), and anterior ocular segment (three eyes). Operative procedures to remove the intraocular foreign bodies (three cases) were guided by the ultrasound biomicroscopy information. CONCLUSIONS: Ultrasound biomicroscopy is a noninvasive method for detecting anterior segment intraocular foreign bodies after perforating trauma. It can be used to accurately diagnose foreign bodies and assist in surgical management, particularly when direct visualization is obscured because of the trauma. In eyes with partial-thickness corneoscleral lacerations or sealed full-thickness corneoscleral laceration and suspected anterior ocular segment foreign body, ultrasound biomicroscopy is a safe and effective method for detecting and localizing foreign bodies in the anterior ocular segment. PMID- 9727512 TI - Combined effect of dorzolamide and latanoprost on the rate of aqueous humor flow. AB - PURPOSE: To determine whether latanoprost, an ocular hypotensive agent believed to enhance uveoscleral outflow of aqueous humor, augments the aqueous-suppressing effect of dorzolamide, a topical carbonic anhydrase inhibitor. METHODS: Twenty four normal subjects underwent measurement of aqueous humor flow by fluorophotometry to determine the flow with placebo, with dorzolamide, and with a combination of dorzolamide and latanoprost. RESULTS: The flow of aqueous humor was suppressed 13% by dorzolamide but not by latanoprost. Latanoprost did not augment the effect of dorzolamide on aqueous humor flow; latanoprost and dorzolamide had additive ocular hypotensive effects. CONCLUSIONS: The uveoscleral flow effect of latanoprost does not improve the aqueous-suppressing effect of dorzolamide, but the two drugs have additive ocular hypotensive effects. PMID- 9727514 TI - Genotype/phenotype correlations in aniridia. AB - PURPOSE: To detect and characterize mutations in cases of familial and sporadic aniridia in Maritime Canada, and to look for indications of genotype/phenotype correlation within the cohort. METHODS: Twelve consecutive and unrelated patients (probands) who had total or nearly complete absence of irides, and four affected relatives, were recruited from Maritime Canada. Clinical data were obtained by chart review and electroretinogram testing. Mutations in the PAX6 gene were detected by single-strand conformation polymorphism and characterized by sequence analysis. RESULTS: Eleven different PAX6 mutations, 10 of which are novel, were found. The four patients with congenital cataracts all had mutations in the C terminal proline-serine-threonine (PST)-rich domain of the PAX6 protein. Electroretinograms of nine of 11 patients displayed depressed scotopic maximum response b-wave amplitudes. The greatest decrease in b-wave amplitudes was seen in patients in whom the paired domain was disrupted by mutation. CONCLUSION: Some aspects of the phenotype of aniridia appear to correlate with the predicted effect of point mutations on the paired and PST domains of the PAX6 protein. PMID- 9727515 TI - Effects of PAX6 mutations on retinal function: an electroretinographic study. AB - PURPOSE: To investigate the retinal function in aniridic patients with documented PAX6 mutations to determine the range of electroretinogram abnormalities in aniridic patients and to relate electroretinogram findings with specific PAX6 mutations. METHODS: Eleven patients with typical aniridia and fully characterized PAX6 mutations underwent electroretinography. RESULTS: In all 11 patients, electroretinogram recordings were abnormal, ranging from mild to severe. Rod related and cone-related activities were equally affected. The amplitude of the oscillatory potentials was the most reduced, followed by the b-wave, then to a milder degree the a-wave. Mutations affecting the paired domain of the PAX6 protein had the biggest impact on the electroretinogram amplitudes. Implicit times were increased in a subgroup with mutations affecting only the homeodomain. CONCLUSION: Patients with aniridia have varying degree of retinal dysfunction, ranging from severely abnormal to almost normal. The paired domain appears to have more impact on retinal function than other regions of the PAX6 protein. It is unclear whether mutations affecting the homeodomain lead to alteration of the photoreceptor function. PMID- 9727516 TI - Laser-induced chorioretinal venous anastomosis for nonischemic central retinal vein occlusion: evaluation of the complications and their risk factors. AB - PURPOSE: To evaluate the complications of laser-induced chorioretinal venous anastomosis in nonischemic central retinal vein occlusion (CRVO) and to identify the associated risks. METHODS: A retrospective consecutive series of 91 eyes (91 patients) with nonischemic CRVO with a mean +/- SD duration of 15.0 +/- 15.2 weeks (range, 3 to 72 weeks )and corrected visual acuity reduced to 20/100 or less because of perfused macular edema were reviewed. All eyes had one or more anastomotic attempts using argon laser (combined with Nd-YAG laser in 46 eyes) and a minimum of 12 months of follow-up. RESULTS: Successful chorioretinal venous anastomoses were created in 49 eyes (54%). Eighteen eyes (20%) had neovascular complications. These consisted of intravitreal, intraretinal, and subretinal neovascular membranes and were significantly associated with retinal ischemia (P < .001). There was avascular fibrous tissue proliferation at the anastomotic site in eight eyes (9%), and it was not associated with retinal ischemia (P = .727). No eye developed further capillary nonperfusion once an anastomosis became functional. A chorioretinal venous anastomosis was associated with improved vision (P < .001); 84% of eyes had an average +/- SD improvement of 4.3 +/- 3.8 lines (range, 2 to 20 lines), with the remaining 16% having no improvement or reduced vision. CONCLUSION: The major vision-threatening complication of laser induced chorioretinal venous anastomosis for nonischemic CRVO is neovascular membranes occurring at the anastomotic site; these are associated with retinal ischemia. Prompt laser photocoagulation to areas of retinal ischemia that develop after the anastomotic attempt has been made may reduce the risk and severity of this complication. PMID- 9727518 TI - Indocyanine green angiographic findings in nonproliferative diabetic retinopathy. AB - PURPOSE: To report angiographic findings of nonproliferative diabetic retinopathy patients by means of indocyanine green angiography. METHODS: Forty-two eyes (42 patients) with nonproliferative diabetic retinopathy were evaluated by indocyanine green angiography in addition to fluorescein angiography. Angiographic findings with the two imaging techniques were compared with the red free fundus appearance. RESULTS: Among the 42 eyes, five (12%) had the appearance on indocyanine green angiography of lobular spotty hyperfluorescent and hypofluorescent areas ("salt and pepper" appearance) in the very late phase. Twenty eyes (48%) presented with diffuse late-phase hyperfluorescence on indocyanine green angiography, corresponding to areas of retinal capillary nonperfusion on fluorescein angiography, and retinal edema. A total of 3,564 microaneurysms were divided into three types: 58 (1.6%) appeared mainly on the fluorescein angiography and very faintly on the indocyanine green angiography, 3,029 (85%) appeared on the fluorescein angiography and the indocyanine green angiography, and 477 (13.4%) were uniquely hyperfluorescent on the indocyanine green angiography. CONCLUSIONS: Indocyanine green angiography in patients with nonproliferative diabetic retinopathy disclosed microvascular findings that were in addition to those shown on fluorescein angiography. These angiographic changes were not observed on fluorescein angiography because of imaging limitations. Indocyanine green angiography may be a useful adjunct to fluorescein angiography in the evaluation of chorioretinal vascular changes in nonproliferative diabetic retinopathy. PMID- 9727517 TI - Recoverin-associated retinopathy: a clinically and immunologically distinctive disease. AB - PURPOSE: To compare the immune response to retinal antigens in a patient with a clinical condition resembling cancer-associated retinopathy with the immune responses of patients with other retinal degenerations or uveitis. METHODS: Cellular and humoral immune responses to retinal S-antigen and recoverin were determined in one patient with disease resembling cancer-associated retinopathy, three patients with other retinal degenerations, and eight patients with uveitis. RESULTS: A cellular immune response against recoverin was found only in the patient with the condition resembling cancer-associated retinopathy. Elevated levels of antibody against recoverin were found in this patient and in one of the three patients with a retinal degeneration, but in none of the eight patients with uveitis. In contrast, moderate lymphocyte responses to retinal S-antigen were found in most of the patients studied, and this response did not distinguish among the patient groups. Levels of serum antibodies against retinal S-antigen were also similar in all patients tested. Serum from the patient with disease resembling cancer-associated retinopathy produced strong immunostaining of the rods, cones, outer plexiform layer, and some cone bipolar cells, but serum from the patients with uveitis or other retinal degenerations did not show specific reactivity with the retina. CONCLUSIONS: We propose that this immunologically and clinically distinctive condition be termed recoverin-associated retinopathy, and we suggest that a cellular immune response against recoverin may be a distinguishing feature of the disorder. PMID- 9727519 TI - Choroidopathy after blunt trauma to the eye: a fluorescein and indocyanine green angiographic study. AB - PURPOSE: To describe the uses of fluorescein angiography and indocyanine green angiography in highlighting traumatic choroidopathy. METHODS: In a prospective study, 21 patients (21 eyes) who presented with traumatic retinal opacity ophthalmoscopically underwent fluorescein angiography and indocyanine green angiography. With indocyanine green angiography, the subtraction method was used for detailed examination. RESULTS: In 11 of 21 eyes, fluorescein angiography showed no abnormalities. On indocyanine green angiography, delayed filling in the choroid was found locally in nine of these 11 eyes. Delayed filling of the choroidal veins was clearly visualized by the subtraction method. In six eyes, intrachoroidal indocyanine green leakage around the choroidal vessels, including vortex veins, was found. In the remaining 10 of the 21 eyes, fluorescein angiography showed fluorescein leakage or a salt-and-pepper appearance in the region of retinal opacity. On indocyanine green angiography, a triangular-shaped area of delayed filling of the choroidal arteries was observed in four eyes. Delayed filling of the choroidal veins was visualized in all 10 eyes, intrachoroidal indocyanine green leakage was found in eight, and dilation and/or narrowing of the choroidal veins and changes in choroidal vasculature were visualized in five eyes. Furthermore, in regions with fluorescein leakage, indocyanine green leakage or hypofluorescence was observed, suggesting damage to the choriocapillaris. CONCLUSION: Indocyanine green angiography allows the analysis of various degrees of choroidal vascular damage. Because the choroidal circulation nourishes the outer retinal layer, traumatic choroidopathy may play a role in the prognosis for visual recovery in eyes affected by contusion retinal opacities. PMID- 9727520 TI - Proton beam irradiation of choroidal hemangiomas. AB - PURPOSE: To present a large series of choroidal hemangiomas treated with proton beam irradiation and to describe the treatment outcomes. METHODS: We treated 54 eyes of 53 patients with choroidal hemangioma. The lesions consisted of 48 circumscribed hemangiomas and six diffuse hemangiomas in patients with Sturge Weber syndrome. The total applied dose was 27.3 Gy in four eyes, 22.7 Gy in three eyes, and 16.4 Gy to 18.2 Gy in 47 eyes. RESULTS: The retina reattached within six months after treatment in all 54 eyes and no recurrence of the secondary retinal detachment occurred within the follow-up period of 6 months to 9 years. Tumors treated with the higher doses regressed faster than tumors treated with the lower doses, but radiation-induced complications of the optic nerve appeared in all four eyes treated with a total dose of 27.3 Gy. Of 31 eyes treated with 16.4 to 18.2 Gy and followed for more than 1 year, 22 had an improvement in their visual acuity, and nine retained the same visual acuity. At the last follow-up examination, the best-corrected visual acuity was 20/20 or better in nine eyes, 20/40 to 20/25 in 13 eyes, 20/100 to 20/50 in six eyes, and 20/200 or less in three eyes. CONCLUSIONS: Proton beam irradiation of choroidal hemangiomas appears to be a valid therapeutic alternative. A total proton dose ranging from 16.4 to 18.2 Gy applied in four daily fractions seems adequate to ensure local control of both tumor and secondary retinal detachment. PMID- 9727521 TI - Second nonocular tumors in survivors of heritable retinoblastoma and prior radiation therapy. AB - PURPOSE: The principal objectives of this study were to estimate the incidence of second tumors among children treated for heritable retinoblastoma during a 50 year period and to investigate the relationship between these tumors and previous radiation therapy. METHODS: The records of all retinoblastoma patients examined at the Mayo Clinic from 1941 through 1990 were retrospectively reviewed. The therapeutic modality used to manage the tumor, the occurrence of any second malignancy, and current follow-up on all patients were evaluated. RESULTS: Eighty two (46%) of 180 children with retinoblastoma had bilateral tumors (76 patients) or unilateral disease and a positive family history (six patients) and were followed for an average of 21.8 years (range, 1 month to 53 years). The Kaplan Meier estimates of second nonocular tumors among the 82 patients with heritable retinoblastoma were 12% at 10 years, 16% at 25 years, and 30% at 40 years. Although 14 of the 15 patients who developed second malignancies had received radiation therapy, only four of the malignancies occurred within the field of irradiation. CONCLUSIONS: The relatively low incidence of second tumors among long-term survivors of heritable retinoblastoma in this series of patients occurred predominantly outside the field of irradiation. The variable incidence of second nonocular malignancies in previous reports may reflect variations in radiation technique and dosage. PMID- 9727522 TI - Treatment of experimental Staphylococcus epidermidis endophthalmitis with oral trovafloxacin. AB - PURPOSE: To investigate the ocular pharmacokinetics and efficacy of oral trovafloxacin, a novel fluoroquinolone antibiotic, in Staphylococcus epidermidis endophthalmitis. METHODS: Albino rabbits (n = 20) were infected with an intravitreal inoculum of S epidermidis (1.0 x 10(8) colony-forming units [CFU/0.1 ml) and 24 hours later received a single oral dose of trovafloxacin (250 mg/kg). Serum and intraocular samples from infected and control (noninfected) eyes were obtained up to 24 hours after antibiotic administration for measurement of trovafloxacin levels. A second group of rabbits (n = 72) was infected intraocularly and randomized 24 hours later to oral trovafloxacin (250 mg/kg twice a day) for 6 days or no treatment (control). Treatment efficacy was assessed by vitreous culture, clinical examination, and histopathology. RESULTS: Following a single dose of trovafloxacin, maximal vitreous levels were achieved at 8 hours in infected eyes, with a penetration ratio of 36%. Vitreous levels were greater than 15 times the minimum inhibitory concentration of the strain employed. In animals with established endophthalmitis, treated eyes were sterilized after 5 days (P = .0495) compared with control eyes, which autosterilized at 14 days. Clinical and histologic examination revealed significant amelioration of anterior segment inflammation in treated eyes, although severe destruction of posterior segment structures occurred in both groups after 6 days of therapy. CONCLUSIONS: These data support trovafloxacin as a potential oral agent for treatment or prophylaxis of S epidermidis endophthalmitis, although retinal alterations that occur over the period required for vitreous sterilization suggest that it will not replace intravitreal therapy in established endophthalmitis. PMID- 9727523 TI - The role of transcranial Doppler in nonarteritic ischemic optic neuropathy. AB - PURPOSE: To investigate through the use of cerebral Doppler technology whether emboli are a more common cause of nonarteritic anterior ischemic optic neuropathy (NAION) than previously recognized. METHODS: Eleven patients with a recent (<121 days) history of nonarteritic anterior ischemic optic neuropathy and 10 age matched controls (event > 121 days) were examined using a Nicolet Pioneer 2020 transcranial Doppler (TCD) unit with a 2-MHz bilateral continuous monitoring capability. The right and left middle cerebral arteries were evaluated simultaneously for 30 minutes at a depth of 50 to 55 mm, and the number of emboli, blood flow velocities, and pulsatility indices were recorded. Data were stored by computer, and hard-copy color recordings were made. RESULTS: None of 11 patients with a recent history of NAION demonstrated microemboli by TCD examination. One patient in the control group who had a remote history of NAION had a microembolic event rate of 12 per hour (six over 30 minutes). This patient had a history of prosthetic cardiac valve replacement and was taking anticoagulation medication at the time of the examination. CONCLUSIONS: Our study did not reveal an increased incidence of embolic events in patients with NAION when they were examined in a transcranial Doppler study of the middle cerebral arteries. This study does not support embolism as a frequent cause of NAION. PMID- 9727524 TI - Relative afferent pupillary defects in patients with Leber hereditary optic neuropathy and unilateral visual loss. AB - PURPOSE: It has been suggested that the pupillary light reaction is relatively preserved in the affected eyes of patients with Leber hereditary optic neuropathy (LHON). To test the hypothesis that visual-pupillomotor dissociation exists in LHON, we performed a retrospective study to evaluate the magnitude of the relative afferent pupillary defect (RAPD) in patients who had experienced monocular visual loss. We also compared the size of the measured RAPD with the size of the RAPD that would be expected on the basis of documented visual field loss. METHODS: We identified a cohort of patients with LHON and monocular visual loss, whose pupillary reactions had been quantified using neutral density filters. From a review of the case records, we determined whether an RAPD was present, as well as the magnitude of the documented RAPDs. We also calculated the expected size of the RAPD for each patient, using previously established templates that correlated the size of the RAPD with the degree of visual field loss. RESULTS: An RAPD was identified in all 10 patients in this study. There was no significant difference between the size of the measured and predicted RAPD, nor did the size of the RAPD correlate with visual acuity or the time interval between the onset of visual loss and evaluation. CONCLUSION: The results of this study do not support the hypothesis that visual-pupillomotor dissociation is a common feature of LHON. PMID- 9727525 TI - Cancer-associated retinopathy vs recoverin-associated retinopathy. PMID- 9727526 TI - Microcirculation architecture of metastases from primary ciliary body and choroidal melanomas. AB - PURPOSE: To describe the microcirculation architecture of metastatic choroidal and ciliary body melanoma. METHOD: Histologic sections of 35 metastases from 19 primary melanomas were stained to demonstrate microcirculation. RESULT: The appearance of microcirculatory networks in metastases is independent of the target organ but associated with the size of the metastatic deposit (estimated coefficient = 0.5959; SE = 0.3024; P = .0488). CONCLUSION: The microcirculatory patterns of primary uveal melanomas that are associated with metastatic behavior appear in foci of metastasis, regardless of the site of dissemination. PMID- 9727527 TI - Ocular manifestations in multiple endocrine neoplasia type 2b. AB - PURPOSE: To describe the clinical and laboratory findings in a patient with multiple endocrine neoplasia type 2b. METHOD: Case report. An 8-year-old boy underwent ophthalmic examination, genetic evaluation, total thyroidectomy, and biopsy of a tongue nodule. RESULTS: Ocular features, including previously unreported iris changes, and their probable origin are discussed. Genetic testing detected the point mutation at codon 918 within the RET protooncogene on chromosome 10, characteristic of multiple endocrine neoplasia type 2b. Histologic analysis of excised thyroid tissue disclosed medullary carcinoma. A tongue nodule proved to be neuromatous. CONCLUSION: Ophthalmologists can play an important role in the recognition of multiple endocrine neoplasia type 2b, a potentially lethal condition. PMID- 9727528 TI - Bilateral secondary angle-closure glaucoma as a complication of anticoagulation in a nanophthalmic patient. AB - PURPOSE: To describe bilateral hemorrhage of the posterior segment and secondary angle-closure glaucoma as sequelae of anticoagulation therapy in a nanophthalmic patient. METHODS: An 80-year-old man who was nanophthalmic and was undergoing anticoagulation therapy presented with declining visual acuity in left eye. Six months later, he experienced declining visual acuity in his right eye. RESULTS: In the LE and six months later in the RE, ocular examination disclosed angle closure glaucoma and a hemorrhagic retinal detachment. Peripheral iridoplasty successfully treated the initial attack. The subretinal hemorrhage was successfully drained by pars plana vitrectomy, retinotomy, and air-fluid exchange in the left eye. Anatomic success and intraocular pressure control were obtained, but visual recovery was limited. CONCLUSION: Intraocular hemorrhage and angle closure glaucoma are potential complications of anticoagulation therapy in a patient with nanophthalmos. PMID- 9727529 TI - Acute posterior multifocal placoid pigment epitheliopathy with bilateral central retinal vein occlusion. AB - PURPOSE: We examined a patient with acute posterior multifocal placoid pigment epitheliopathy and bilateral central retinal vein occlusion. METHOD: Case report. A 28-year-old woman presented with the typical findings of acute posterior multifocal placoid pigment epitheliopathy. One month after presentation, the patient developed bilateral central retinal vein occlusion. RESULT: Four months after presentation, resolution of the bilateral central retinal vein occlusion resulted in a corrected visual acuity of RE, 20/25 and LE, 3/400. CONCLUSION: Vasculitis, sometimes associated with acute posterior multifocal placoid pigment epitheliopathy, can affect the ciliary vessels that supply the optic disk. This may result in axoplasmic stasis and compression of the central retinal vein, leading to impending or complete central retinal vein occlusion. PMID- 9727530 TI - Familial dysplasminogenemia with central retinal vein and cilioretinal artery occlusion. AB - PURPOSE: To report a 49-year-old woman with unilateral central retinal vein occlusion and ipsilateral cilioretinal artery occlusion who showed familial dysplasminogenemia associated with elevated lipoprotein(a). METHOD: Case report. RESULTS: Extensive hemostatic and coagulation studies for causes of thrombosis, as well as family studies, disclosed decreased plasminogen activity without reduction of plasminogen antigen in the patient, her two siblings, and her two children. The patient also showed elevated lipoprotein(a). CONCLUSION: The combination of decreased plasminogen activity and elevated lipoprotein(a) should be considered as a possible cause of retinal vein and artery occlusion. PMID- 9727531 TI - Vitreitis and Waldenstrom's macroglobulinemia. AB - PURPOSE: To describe vitreitis in a patient with Waldenstrom's macroglobulinemia. METHODS: Case report and review of pertinent literature. RESULTS: A 90-year-old man developed vitreitis 10 years after a systemic diagnosis of a lymphoproliferative disorder. Numerous small, normal-appearing lymphocytes were seen on pathologic examination of the vitreous. He developed worsening lymphadenopathy and was diagnosed with Waldenstrom's macroglobulinemia after systemic review. CONCLUSION: Chronic lymphoproliferative diseases such as Waldenstrom's macroglobulinemia may cause vitreitis. PMID- 9727532 TI - Ptosis in Waldenstrom's macroglobulinemia. AB - PURPOSE: To report a case of chronic, progressive unilateral blepharoptosis in a 73-year-old woman with Waldenstrom's macroglobulinemia. METHOD: Case report. A biopsy was performed on a thickened and indurated tarsal plate that we believed had resulted in mechanical blepharoptosis. RESULTS: Histologic and immunohistochemistry studies of the biopsy specimen demonstrated a lymphoplasmacytoid cell infiltrate with monoclonal antibodies consistent with Waldenstrom's macroglobulinemia. CONCLUSION: Involvement of the tarsal conjunctiva and tarsus in Waldenstrom's macroglobulinemia is a newly recognized cause of eyelid thickening and ptosis. PMID- 9727533 TI - Poststreptococcal uveitis. AB - PURPOSE: To report that uveitis may be a manifestation of poststreptococcal syndrome. METHODS: Case report. Documented attacks of bilateral uveitis were clearly associated with streptococcal infection. RESULTS: Group A streptococcal infection was evident in all bilateral uveitis attacks, which were treated with local or systemic corticosteroids and penicillin. The frequency and severity of the attacks were reduced by penicillin prophylaxis and tonsillectomy. CONCLUSIONS: Uveitis should be included as a possible manifestation of poststreptococcal syndrome. If coexisting streptococcal infection is demonstrated, penicillin prophylaxis should be considered. PMID- 9727534 TI - Optic nerve head neovascularization in a patient with inactive cytomegalovirus retinitis and immune recovery. AB - PURPOSE: To report a case of optic nerve head neovascularization in a patient with acquired immunodeficiency syndrome (AIDS) associated with inactive cytomegalovirus retinitis and immune recovery. METHOD: Case report. RESULTS: We examined a 29-year-old man with AIDS and inactive cytomegalovirus retinitis and found vitritis and prominent optic nerve head neovascularization. The patient had been treated with reverse transcriptase and protease inhibitors, resulting in a notable rise in CD4+ lymphocyte count and an undetectable human immunodeficiency virus (HIV)-RNA viral load. No cause of neovascularization other than intraocular inflammation was detected. CONCLUSION: Immune recovery in a setting of inactive cytomegalovirus retinitis can result in optic nerve head neovascularization, as seen in our patient. PMID- 9727535 TI - Choroidal neovascularization as a late complication of hyperparathyroidism. AB - PURPOSE: To report choroidal neovascularization in a patient with sclerochoroidal calcification secondary to hyperparathyroidism. METHOD: Case report. Bilateral ocular fundus abnormalities observed in a 74-year-old woman were documented with fluorescein angiography and ultrasonography. RESULTS: Two yellow, irregular, oval placoid lesions were observed in the fundus of each eye. Ultrasound examination showed that these areas were extremely sonoreflective and exhibited orbital shadowing. In the left eye, one lesion was accompanied by lipid exudates, and subsequent fluorescein angiography disclosed choroidal neovascularization. Medical history included hyperparathyroidism treated with surgical excision of a parathyroid adenoma 27 years earlier. CONCLUSION: Choroidal neovascularization may be a late complication of hyperparathyroidism. PMID- 9727536 TI - Trochlear nerve enhancement on three-dimensional magnetic resonance imaging in Fisher syndrome. AB - PURPOSE: To examine the lesion associated with external ophthalmoplegia in Fisher syndrome using three-dimensional magnetic resonance imaging (3-D MRI). METHOD: Case report. A 65-year-old woman with Fisher syndrome was investigated by gadolinium-enhanced 3-D MRI. RESULT: The extramedullary portion of the left trochlear nerve was enhanced. CONCLUSION: Contrast-enhanced 3-D MRI revealed that the lesion responsible for the external ophthalmoplegia in Fisher syndrome is located in the extramedullary portion of the trochlear nerve. PMID- 9727537 TI - Primary glaucoma triple procedure in patients with primary open-angle glaucoma. PMID- 9727538 TI - Corneal thickness and curvature in normal-tension glaucoma. PMID- 9727539 TI - Antiviral chemoprophylaxis after occupational exposure to human immunodeficiency virus. PMID- 9727540 TI - Efficacy and safety of the Protek (Vifilcon A) therapeutic soft contact lens. PMID- 9727541 TI - Prospective study of C-reactive protein and the risk of future cardiovascular events among apparently healthy women. AB - BACKGROUND: C-reactive protein (CRP) predicts risk of myocardial infarction (MI) and stroke among apparently healthy men, but in women, virtually no data are available. METHODS AND RESULTS: CRP was measured in baseline blood samples from 122 apparently healthy participants in the Women's Health Study who subsequently suffered a first cardiovascular event and from 244 age- and smoking-matched control subjects who remained free of cardiovascular disease during a 3-year follow-up period. Women who developed cardiovascular events had higher baseline CRP levels than control subjects (P=0.0001), such that those with the highest levels at baseline had a 5-fold increase in risk of any vascular event (RR=4.8; 95% CI, 2.3 to 10.1; P=0.0001) and a 7-fold increase in risk of MI or stroke (RR=7.3; 95% CI, 2.7 to 19.9; P=0.0001). Risk estimates were independent of other risk factors, and prediction models that included CRP provided a better method to predict risk than models that excluded CRP (all P values <0.01). In stratified analyses, CRP was a predictor among subgroups of women with low as well as high risk as defined by other cardiovascular risk factors. CONCLUSIONS: In these prospective data among women, CRP is a strong independent risk factor for cardiovascular disease that adds to the predictive value of risk models based on usual factors alone. (Circulation. 1998;98:731-733.) PMID- 9727542 TI - Randomized, placebo-controlled trial of platelet glycoprotein IIb/IIIa blockade with primary angioplasty for acute myocardial infarction. ReoPro and Primary PTCA Organization and Randomized Trial (RAPPORT) Investigators. AB - BACKGROUND: The benefit of catheter-based reperfusion for acute myocardial infarction (MI) is limited by a 5% to 15% incidence of in-hospital major ischemic events, usually caused by infarct artery reocclusion, and a 20% to 40% need for repeat percutaneous or surgical revascularization. Platelets play a key role in the process of early infarct artery reocclusion, but inhibition of aggregation via the glycoprotein IIb/IIIa receptor has not been prospectively evaluated in the setting of acute MI. METHODS AND RESULTS: Patients with acute MI of <12 hours' duration were randomized, on a double-blind basis, to placebo or abciximab if they were deemed candidates for primary PTCA. The primary efficacy end point was death, reinfarction, or any (urgent or elective) target vessel revascularization (TVR) at 6 months by intention-to-treat (ITT) analysis. Other key prespecified end points were early (7 and 30 days) death, reinfarction, or urgent TVR. The baseline clinical and angiographic variables of the 483 (242 placebo and 241 abciximab) patients were balanced. There was no difference in the incidence of the primary 6-month end point (ITT analysis) in the 2 groups (28.1% and 28.2%, P=0.97, of the placebo and abciximab patients, respectively). However, abciximab significantly reduced the incidence of death, reinfarction, or urgent TVR at all time points assessed (9.9% versus 3.3%, P=0.003, at 7 days; 11.2% versus 5.8%, P=0.03, at 30 days; and 17.8% versus 11.6%, P=0.05, at 6 months). Analysis by actual treatment with PTCA and study drug demonstrated a considerable effect of abciximab with respect to death or reinfarction: 4.7% versus 1.4%, P=0.047, at 7 days; 5.8% versus 3.2%, P=0.20, at 30 days; and 12.0% versus 6.9%, P=0.07, at 6 months. The need for unplanned, "bail-out" stenting was reduced by 42% in the abciximab group (20.4% versus 11.9%, P=0.008). Major bleeding occurred significantly more frequently in the abciximab group (16.6% versus 9.5%, P=0.02), mostly at the arterial access site. There was no intracranial hemorrhage in either group. CONCLUSIONS: Aggressive platelet inhibition with abciximab during primary PTCA for acute MI yielded a substantial reduction in the acute (30-day) phase for death, reinfarction, and urgent target vessel revascularization. However, the bleeding rates were excessive, and the 6-month primary end point, which included elective revascularization, was not favorably affected. PMID- 9727543 TI - Activation of platelets in platelet-rich plasma by rotablation is speed-dependent and can be inhibited by abciximab (c7E3 Fab; ReoPro). AB - BACKGROUND: Rotational atherectomy with the Rotablator catheter has improved percutaneous treatment of certain coronary atherosclerotic lesions, but the "no reflow" phenomenon remains a serious complication. Because platelet activation by rotablation may contribute to the no-reflow phenomenon, we developed an in vitro system to test the effect of rotablation on platelets in the absence or presence of platelet GP IIb/IIIa receptor blockade with abciximab. METHODS AND RESULTS: Platelet-rich plasma (PRP) was prepared from 28 healthy human volunteers. PRP was divided into 4 samples: (1) no treatment, (2) 6D1 (anti-GP Ib), (3) c7E3 Fab (anti-GP IIb/IIIa+alpha(v)beta3), and (4) c7E3 Fab+6D1. Samples were pumped through a flow chamber containing a 2.5-mm burr rotating at various speeds and then placed in an aggregometer. PRP samples tested in the absence of antibody underwent more rapid and extensive aggregation when rotablated at 150000 and 180000 rpm compared with 0 rpm (P<0.001 at both speeds). Preincubation of platelets with c7E3 Fab decreased the slope of aggregation at each rotablation speed, with 98%, 79%, and 71% reductions at 70000, 150000, and 180000 rpm, respectively (P=0.09 for 70000 and P<0.001 for both 150000 and 180000 rpm). Preincubation of platelets with 6D1 did not decrease the slope of aggregation at any rotablation speed (P>0.5, P=0.99, and P=0.091 for 70000, 150000, and 180000 rpm). Platelet ATP release, a marker of granule release and cell damage, was markedly increased at 180000 rpm (P=0.002 compared with 0 rpm in the control group). Electron microscopy revealed extensive rotablation-induced platelet damage at 150000 and 180000 rpm, and leakage of LDH confirmed platelet lysis at these speeds (P=0.002 and P<0.001 compared with 0 rpm). CONCLUSIONS: High-speed rotablation induces platelet activation of PRP, leading to aggregation; pretreating PRP with abciximab decreases the aggregation. These data suggest that pretreatment of patients with abciximab may decrease rotablation-induced platelet aggregation during rotational atherectomy. PMID- 9727544 TI - Randomized, double-blind crossover study to investigate the effects of amlodipine and isosorbide mononitrate on the time course and severity of exercise-induced myocardial stunning. AB - BACKGROUND: Myocardial stunning may cause prolonged left ventricular dysfunction after exercise-induced ischemia that can be attenuated by calcium antagonists in animal models. To assess their effects in humans, we performed a randomized, double-blind crossover study comparing the calcium antagonist amlodipine (10 mg once daily) versus isosorbide mononitrate (ISMN, 50 mg once daily) on postexercise stunning. METHODS AND RESULTS: Twenty-four men with chronic stable angina and normal left ventricular function underwent serial quantitative exercise stress echocardiography after 3 weeks on each treatment to assess the degree of postexercise stunning with simultaneous sestamibi single-photon emission computed tomography perfusion scans at peak stress to quantify the ischemic burden. Exercise time (P=1), maximum ST depression (P=0.48), and sestamibi single-photon emission computed tomography scores (P=0.17) were unchanged between treatments. Stunning occurred more often with ISMN than amlodipine (82% versus 48%). The global and segmental stress echocardiography parameters of stunning were attenuated in patients while taking amlodipine compared with ISMN. Shortening fractions and ejection fractions were less impaired 30 minutes after exercise in patients receiving amlodipine (3.5+/-1.4% versus 2.5+/-1.4%, P=0.014, and 59.7+/-5.4% versus 54.5+/-8%, P<0.001); similarly, the isovolumic relaxation period was less prolonged with amlodipine (93+/-15.5 versus 106.3+/-14.9 ms, P=0.018). CONCLUSIONS: Despite comparable levels of ischemia, amlodipine attenuated stunning when compared with ISMN. This beneficial effect may relate to a prevention of the calcium overload implicated in the pathogenesis of stunning. PMID- 9727545 TI - Acute hemodynamic interaction of aspirin and ticlopidine with enalapril: results of a double-blind, randomized comparative trial. AB - BACKGROUND: Coprescription of aspirin and ACE inhibitors is frequent in heart failure caused by coronary artery disease. Negative interaction between aspirin and enalapril has been reported, presumably through inhibition by aspirin of ACE inhibitor-induced prostaglandin synthesis. Ticlopidine is a potent antiplatelet agent without interaction with prostaglandin synthesis. METHODS AND RESULTS: The objective of this study was to compare the influence of a coadministration of ticlopidine or aspirin on the hemodynamic effects of an ACE inhibitor (enalapril) in patients with chronic heart failure. Twenty patients with severe heart failure were enrolled in a double-blind comparative trial and allocated to ticlopidine (500 mg daily, 12 patients) or aspirin (325 mg daily, 8 patients). Hemodynamic evaluation was performed after 7 days of treatment, every hour for 4 hours after an oral administration of 10 mg of enalapril. Significant reductions in systemic vascular resistance were observed in the ticlopidine group, in contrast to no significant decrease in the aspirin group. A significant (P=0.03) time-by treatment interaction indicated significant aspirin-enalapril drug interaction. Total pulmonary resistance decreased significantly in both groups, with no difference between patients assigned to aspirin or ticlopidine. CONCLUSIONS: Enalapril reduced systemic vascular resistance more effectively when given in combination with ticlopidine than with aspirin. In contrast, the reduction in total pulmonary resistance is similar when enalapril is administered in combination with aspirin or ticlopidine. Negative aspirin-enalapril interaction on prostaglandin synthesis presumably alters vasodilatation in systemic vessels, whereas prostaglandin-independent actions of ACE inhibition such as pulmonary arterial vasodilatation are maintained. PMID- 9727546 TI - Accuracy and impact of presumed cause in patients with cardiac arrest. AB - BACKGROUND: International guidelines recommend differentiation between cardiac and noncardiac causes of cardiac arrest. The aim of this study was to find the rate of agreement between primarily postulated and definitive causes of cardiac arrest. METHODS AND RESULTS: We retrospectively analyzed the primarily presumed cause of cardiac arrest as determined by the emergency room physician on admission in all patients admitted to the emergency department of one urban tertiary care hospital. This was compared with the definitive cause as established by clinical evidence or autopsy. Within 4 years, the initially presumed cause was unclear in 24 (4%) of 593 patients. In the remaining 569 patients, the presumed cause was correct in 509 (89%) and wrong in 60 (11%) cases. Cardiac origin was presumed in 421 (71%) and the definitive cause in 408 (69%) cases. Noncardiac origin was presumed in 148 (25%) and the definitive cause in 185 (31%) patients. Presumed cardiac cause was sensitive (96%) but less specific (77%). Noncardiac causes such as pulmonary embolism, cerebral disorders, or exsanguination were those most frequently overlooked. Asystole occurred significantly more often in patients in whom presumed cause remained undetermined or differed from the definitive cause. CONCLUSIONS: Cause of cardiac arrest is not as easily recognized as anticipated, especially when the initial rhythm is different from ventricular fibrillation. This might affect comparability of study results, therapeutic strategies, prognosis, and outcome. Patients in whom the presumed cause was confirmed as being correct had significantly better survival and neurological outcome. PMID- 9727547 TI - Sympathetic activity in obese subjects with and without obstructive sleep apnea. AB - BACKGROUND: Obese humans are reported to have increased muscle sympathetic nerve activity (MSNA). Obstructive sleep apnea (OSA) may also be accompanied by increased MSNA. Because there is a high prevalence of OSA in obese humans, it is possible that high MSNA reported in obese subjects may in fact reflect the presence of OSA in these subjects. We tested the hypothesis that obesity, per se, in the absence of OSA, is not accompanied by increased MSNA. METHODS AND RESULTS: We measured MSNA in 25 healthy normal-weight subjects and 30 healthy sedentary obese subjects. All subjects were screened by history and examination to exclude subjects with OSA or hypertension. OSA was further excluded by overnight polysomnographic studies. Despite careful screening, polysomnography revealed that 1 of 25 normal-weight subjects and 9 of 30 obese subjects had occult OSA (P=0.015). MSNA was similar in normal-weight subjects (41+/-3 bursts per 100 heartbeats) and obese subjects without sleep apnea (42+/-3 bursts per 100 heartbeats, P=0.99). MSNA in the 9 obese subjects with occult OSA was 61+/-8 bursts per 100 heartbeats, which was higher than MSNA in normal-weight subjects without sleep apnea (P=0.02) and higher than MSNA in obese subjects without sleep apnea (P=0.02). CONCLUSIONS: Obesity alone, in the absence of OSA, is not accompanied by increased sympathetic activity to muscle blood vessels. PMID- 9727548 TI - Idiopathic dilated cardiomyopathy: a superantigen-driven autoimmune disease. AB - BACKGROUND: Many cases of idiopathic dilated cardiomyopathy (IDC) result from an inflammatory myocarditis. The specific immunological mechanisms are not yet defined. Various autoimmune diseases are associated with superantigen-triggered immune responses, resulting in massive T-cell activation and tissue damage. We studied 3 cases in a search for evidence that such a phenomenon is also implicated in IDC. METHODS AND RESULTS: Myocardial, lymph node, and thymic tissue samples were obtained from IDC patients who were undergoing heart transplantation. Infiltrating immune-cell phenotypes and gene expression of T cell receptor (TCR) alpha- and beta-chain variable (Valpha and Vbeta) regions were analyzed by immunostaining and polymerase chain reaction. Similar technical approaches were used to assay the tissues for the presence of coxsackievirus B (CVB). In all the specimens analyzed, an overexpression of the TCR Vbeta3, Vbeta7, and Vbeta13.1 gene families was detected among the infiltrating T cells. These tissues were also found to be CVB3-positive. In vitro exposure of peripheral blood mononuclear cells to lysates of cells infected with CVB3 was capable of stimulating expansion of the same TCR Vbeta families. The TCR Valpha repertoire was never found to be skewed. CONCLUSIONS: A superantigen-mediated immune response is involved in human heart disease. CVB3 may directly or indirectly trigger this response, suggesting a possible mechanistic link between CVB infection and myocarditis development progressing to IDC. PMID- 9727549 TI - Preformed IgG antibodies against major histocompatibility complex class II antigens are major risk factors for high-grade cellular rejection in recipients of heart transplantation. AB - BACKGROUND: Preformed anti-HLA antibodies reacting specifically with donor lymphocytes have been associated with acute vascular rejection and early cardiac allograft failure. However, the effect of preformed anti-HLA antibodies directed against allogeneic major histocompatibility complex (MHC) class I or II antigens of a donor panel on heart transplantation outcome has not been extensively studied. METHODS AND RESULTS: The study group consisted of 68 patients who received cardiac transplants between 1989 and 1996 and who were at high risk for developing anti-HLA antibodies before transplantation. The effect of preformed antibodies against allogeneic MHC class I or class II antigens on the development of early high-grade cellular rejection and on cumulative annual rejection frequency was determined. Both patients with left ventricular assist devices and retransplantation candidates had a similar increase in the frequency of IgG anti MHC class II antibodies (IgG anti-II) compared with control subjects (P<0.0001), whereas the frequency of IgG anti-MHC class I antibodies (IgG anti-I) was elevated only in patients with left ventricular assist devices. Pretransplantation IgG anti-II predicted early development of high-grade cellular rejection (P=0.006) and higher cumulative annual rejection frequency (P<0.001) in both of these sensitized patient groups. Among retransplantation recipients, a match between donors 1 and 2 at HLA-A additionally predicted an earlier time to a high-grade cellular rejection. CONCLUSIONS: These results emphasize the importance of specifically screening heart transplantation candidates for the presence of IgG antibodies directed against MHC class II molecules and suggest that strategies aimed at their reduction may have an impact on the onset and frequency of high-grade cellular rejections after transplantation. PMID- 9727550 TI - Inhibitory effects of antioxidants on neonatal rat cardiac myocyte hypertrophy induced by tumor necrosis factor-alpha and angiotensin II. AB - BACKGROUND: Tumor necrosis factor-alpha (TNF-alpha) and angiotensin II (Ang II) modulate heart failure in part by provoking the hypertrophic response. Signal transduction pathways of those factors are implicated in reactive oxygen intermediates (ROIs). Therefore, we hypothesized that TNF-alpha and Ang II might cause myocyte hypertrophy via the generation of ROIs. METHODS AND RESULTS: To test the hypothesis, we tested whether TNF-alpha and Ang II could induce the generation of ROIs and whether antioxidants such as butylated hydroxyanisole (BHA), vitamin E, and catalase might inhibit the hypertrophy in cultured neonatal rat cardiac myocytes. ROIs were measured by the ROI-specific probe 2',7' dichlorofluorescin diacetate in cultured cardiac myocytes. We demonstrated that TNF-alpha and Ang II induced the generation of ROIs in a dose-dependent manner. TNF-alpha (10 ng/mL) and Ang II (100 nmol/L) enlarged cardiac myocytes and increased [3H]leucine uptake, and BHA (10 micromol/L) significantly inhibited both effects. Other antioxidants, such as vitamin E (1 microg/mL) and catalase (100 U/mL), also inhibited the enlargement of cardiac myocytes induced by TNF alpha. CONCLUSIONS: These results indicate that TNF-alpha and Ang II cause hypertrophy in part via the generation of ROIs in cardiac myocytes. PMID- 9727551 TI - Evidence that macrophages in atherosclerotic lesions contain angiotensin II. AB - BACKGROUND: We have reported that human mononuclear leukocytes contain large amounts of angiotensin II (Ang II). The goal of the present study was to test the hypothesis that Ang II is present in monocyte/macrophages in atherosclerotic lesions. METHODS AND RESULTS: Segments of thoracic aorta and left circumflex coronary artery were obtained from 3 groups of cynomolgus monkeys: normal, atherosclerotic, and regression. Samples of human coronary arterial atherosclerotic lesions were obtained from directional atherectomy. Sections were stained for Ang II with 3 different polyclonal rabbit anti-human Ang II antisera. In aorta and coronary arteries from normal monkeys, there was no or minimal anti Ang II staining in endothelial cells. All sections from atherosclerotic monkeys displayed discrete, localized regions of staining for Ang II in intima-media. Macrophages were present throughout the atherosclerotic intima-media, and anti Ang II staining appeared to colocalize with macrophages. All human coronary atherectomy samples stained positive for Ang II and macrophages. Staining for both Ang II and macrophages was observed in vascular lesions from all 5 monkeys after regression of atherosclerosis, but staining was less extensive than in atherosclerotic blood vessels from monkeys. CONCLUSIONS: These findings suggest that Ang II is present in atherosclerotic lesions in monkeys and humans, colocalizes with macrophages in intima-media of atherosclerotic vessels from monkeys, and decreases in lesions in monkeys with regression of atherosclerosis. PMID- 9727552 TI - Improved internal defibrillation success with shocks timed to the morphology electrogram. AB - BACKGROUND: A previous retrospective study by our group suggested that shocks timed to the upslope of the shocking lead electrogram improved defibrillation efficacy. The goal of this study was to prospectively determine whether defibrillation threshold could be reduced by use of an algorithm that timed shocks to the upslope of coarse ventricular fibrillation (test treatment) compared with shocks delivered asynchronously after 10 seconds of fibrillation (control treatment). METHODS AND RESULTS: Ten pigs were instrumented with a 3 lead system for internal defibrillation. Initial estimates of the energy required to achieve defibrillation E50 for both treatments were made by an up/down method. Subsequently, additional shocks at V50+/-10% and V50-20% were given for each treatment to obtain data points at higher and lower intensities. Probability-of success curves were estimated for both treatments by the best-fit method. Energies required were significantly lower for the timed shocks than for the asynchronous shocks (P<0.00 1). E80 was reduced 15.5%, from 27.1+/-2.5 to 22.9+/ 1.8 J (P<0.002). The width of the probability-of-success curve (E80-E20) for the test treatment was also significantly narrower than that for the control treatment (7.1+/-0.9 versus 10.8+/-1.7, P<0.01). Normalized curve width (E80 E20)/E50 was decreased from 51+/-5% of E50 for control shocks to 37+/-4% of E50 for synchronous shocks (P<0.02). CONCLUSIONS: In this model, defibrillation threshold is lower and more deterministic when shocks are timed to the upslope of the shocking lead electrogram. If a similar reduction is observed in humans, shock timing may lower defibrillation threshold and simplify programming of shock intensity. PMID- 9727553 TI - Protective effect of oral xemilofiban in arterial thrombosis in dogs: increased activity in combination with aspirin. AB - BACKGROUND: Inhibition of platelet aggregation by preventing the binding of fibrinogen to glycoprotein (GP) IIb/IIIa on activated platelets results in antithrombotic activity. We report on the antithrombotic effect of xemilofiban (SC-54684A), an oral GP IIb/IIIa antagonist, administered alone or with aspirin (ASA) in an acute thrombosis model. METHODS AND RESULTS: Conscious dogs were treated with xemilofiban (1.25, 2.5, 5.0, or 6 mg/kg, n=6); low-dose (LD, 81 mg) ASA, n=7; high-dose (HD, 162 mg) ASA, n=6; xemilofibran 1.25 mg/kg plus LD ASA, n=6; xemilofibran 1.25 mg/kg plus HD ASA, n=6; or placebo, n=7. Dogs were anesthetized 60 minutes later, and the effects of the treatments were evaluated after electrolytic injury (250 microA for 180 minutes) in the left circumflex coronary artery. Bleeding time (BT) was assessed in a separate study. Incidence of thrombosis was reduced (P<0.05) by xemilofiban > or =2.5 mg/kg, HD ASA, or xemilofiban 1.25 mg/kg plus HD ASA compared with placebo. Xemilofiban > or =2.5 mg/kg or xemilofiban 1.25 mg/kg plus HD ASA significantly increased time to occlusion, inhibited ex vivo platelet aggregation to collagen >90%, and prevented or decreased (P<0.05) cyclic flow variations (CFVs) compared with placebo. BT was increased (P<0.05) with xemilofiban > or =2.5 mg/kg but not with xemilofiban 1.25 mg/kg plus HD ASA. CONCLUSIONS: Xemilofiban > or =2.5 mg/kg, HD ASA, or xemilofiban 1.25 mg/kg plus HD ASA significantly reduced the incidence of thrombosis. These doses of xemilofiban or xemilofiban 1.25 mg/kg plus HD ASA increased time to occlusion, inhibited ex vivo platelet aggregation by >90%, and prevented or reduced CFVs. Xemilofiban > or =2.5 mg/kg but not xemilofiban 1.25 mg/kg plus HD ASA significantly increased BT. PMID- 9727554 TI - Images in cardiovascular medicine. Myocardial bridging. PMID- 9727555 TI - Images in cardiovascular medicine. Vasoconstriction after coronary stenting. PMID- 9727556 TI - Noncardiac surgery in CAD patients. PMID- 9727557 TI - Adenosine-induced preconditioning of human myocardium? PMID- 9727558 TI - Treatment of pulmonary embolism with thrombolysis. PMID- 9727559 TI - Platelet Pl(A) polymorphism, myocardial infarction, and extent of coronary artery disease. PMID- 9727560 TI - Lack of correlation between V3-loop peptide enzyme immunoassay serologic subtyping and genetic sequencing. AB - OBJECTIVE: To compare the performance of V3-loop peptide enzyme immunoassay (PEIA) methodologies from four different laboratories for subtyping HIV-1, and to determine the causes for the lack of correlation between V3-loop PEIA serotyping and subtyping by sequencing. MATERIALS AND METHODS: Synthetic peptides derived from the amino-acid consensus sequences of the V3-loop of group M strains representing genetic subtypes A-F as well as reference strains were evaluated in PEIA by four different laboratories for their ability to accurately determine the subtype in a panel of 85 sera obtained from persons infected with known HIV-1 subtypes (28 subtype A, 34 subtype B, four subtype C, 10 subtype D, seven subtype F, one each of subtype H and G). Furthermore, the V3 loop of the corresponding virus was compared with the V3 loop of the peptides used in PEIA. RESULTS: The correlation between HIV-1 subtyping by sequencing and V3-loop PEIA from the different laboratories varied considerably for the different HIV-1 subtypes: subtype A (46-68%), B (38-85%), C (75-100%), D (29-50%), and F (17-57%). A 70% agreement between PEIA and sequencing subtypes was observed for samples with the concordant presence of the same octameric sequences in the V3 loop of the virus and the V3 loop of the peptide used in PEIA; however, only 42% of specimens with different V3-loop octameric viral and peptide sequences yielded concordant results in V3-loop serotyping and genetic subtyping. CONCLUSION: Our results indicate that V3-loop PEIA methodologies used in different laboratories correlate poorly with genetic subtyping, and that their accuracy to predict HIV-1 subtypes in sera of Belgian individuals infected with different HIV-1 subtypes (A, B, C, D, F, G and H) vary considerably. The poor correlation between serotyping and genetic subtyping was partly due to the simultaneous occurrence of subtype specific octameric sequences at the tip of the V3 loop of viruses belonging to different genetic subtypes. PMID- 9727561 TI - Structure-based design of peptides that recognize the CD4 binding domain of HIV-1 gp120. AB - DESIGN: Envelope protein-specific antiviral peptides, called mucibodies, that can specifically recognize and bind to the surface unit protein gp120 of HIV-1 were designed. The initial mucibody binding target was the V3 loop of HIV-1 gp120. Here, the gp120-CD4 binding domain was chosen as the site of mucibody binding. The CD4 binding domain of gp120 is known to be a conformational epitope and is involved in the earliest events of viral entry into many cells. METHODS: The design of the mucibody antivirals was based on previous observations that antibody complementarity determining regions (CDR) are generally similar to the repeating loops or knob structures found in the 20-residue tandem repeat domain of human mucin MUC1. The heavy chain CDR3 from the bacteriophage display antibody b12 was used to construct two mucibodies, b12-CDR1 and b12-26. RESULTS: Peptides corresponding to three tandem repeats were shown to bind directly to the CD4 binding domain of HIV-1 gp120 in a solid-phase enzyme-linked immunosorbent assay. These mucibody peptides also disrupted the gp120-CD4 interaction in a solution phase inhibition assay. Finally, mucibodies neutralized primary and laboratory macrophage-tropic isolates of HIV-1. CONCLUSIONS: There is a potential for medical use of these peptides as topical vaginal microbicides in preventing HIV-1 transmission during sexual contact. These results also suggest that multivalent, non-immunogenic binding proteins of virtually any specificity could be constructed for use in therapeutic applications involving infectious diseases and immune system dysfunction. PMID- 9727562 TI - Improved detection of HIV-2 proviral DNA in dually seroreactive individuals by PCR. AB - OBJECTIVE: To improve the detection rate of HIV-2 proviral DNA in primary uncultured peripheral blood mononuclear cells (PBMC) of HIV-2-seroreactive and HIV-1-HIV-2 dually seroreactive individuals. MATERIALS AND METHODS: Two newly designed HIV-2 PCR primer pairs in the long terminal repeat (LTR) gag and gag-pol regions and a previously described env and LTR HIV-2 PCR primer pairs were tested on samples from 66 confirmed HIV-2-seropositive individuals (The Gambia, 40; Cote d'Ivoire, 17; Guinea-Bissau, nine), 209 dually seroreactive individuals (The Gambia, 82; Cote d'Ivoire, 127), 24 genetically characterized isolated HIV-1 strains (group M subtypes A-H and group O), one simian immunodeficiency virus (SIV) strain cpz, 10 HIV-2 isolates (subtype A, B and unidentified), two SIVsm isolates, and 10 seronegative samples. RESULTS: All HIV-2 primers evaluated showed 100% specificity since there was no amplification observed with 24 HIV-1, one SIVcpz and 10 seronegative samples. One single copy of the HIV-2 genome could be detected with all outer primer pairs as well as all inner primer pairs on one PCR round used. Sensitivity of primers (at least one of the four primer pairs was positive) to HIV-2-seropositive samples was 100% (all nine) in Guinea-Bissau, 71% (12/17) in Cote d'Ivoire, 100% (all 20) in Gambian AIDS patients, and 85% (17/20) in Gambian pregnant women. Doubling the PBMC of dually seroreactive individuals from 7.5 x 10(4) to 1.5 x 10(5) in the PCR revealed the presence of both HIV-1 and 2 proviral DNA in 72% (92/127) in Cote d'Ivoire and 72% (59/82) in The Gambia. By doubling the number of PBMC, HIV-2 detection in dually seroreactive individuals by PCR was increased from 65 to 77% in Cote d'Ivoire and from 67 to 83% in The Gambia. CONCLUSIONS: The use of 1.5 x 10(5) primary uncultured PBMC and the newly designed HIV-2 primer pairs allowed us to document the highest percentage (72%) ever reported of HIV-1-HIV-2 dual infections amongst HIV-1-HIV-2 dually seroreactive individuals in Cote d'Ivoire and The Gambia. Improved detection of HIV-2 proviral DNA, rather than exposure to both viruses, infection with only one virus, or infection with a unique third virus containing epitopes common to both HIV-1 and HIV-2, contributes to a more accurate monitoring of the prevalence of HIV-1-HIV-2 dual infections. PMID- 9727563 TI - Cytotoxic T-cell responses to HIV-1 reverse transcriptase, integrase and protease. AB - OBJECTIVES: To determine immunodominant regions and new epitopes for cytotoxic T cells (CTL) directed against the HIV-1 pol products reverse transcriptase (RT), integrase and protease in a large cohort of patients at different stages of disease. DESIGN AND METHODS: Cross-sectional analysis of 98 patients from the French IMMUNOCO cohort (CD4 counts: 125-1050 x 10(6) cells/l), monitored for CTL recognition of HIV-1 pol products using recombinant vaccinia virus constructs and synthetic peptides. RESULTS: Memory CTL responses against HIV-1 pol products were detected in 78% of all patients whatever the stage of disease. RT was more immunogenic (81%, 30 out of 37 patients) than integrase and protease (51% and 24%, respectively). CTL recognition of RT was more frequent against Pol amino acids 310-460 (61%, 11 out of 18 patients) than against the other three portions (Pol 168-310, Pol 450-600, Pol 590-728) in patients with CD4 counts > 400 x 10(6)/l, whereas in patients at advanced stages no prominent differences were observed. Two new clusters of antigenic regions were found in the NH2 segment: three epitopes between amino-acids Pol 200 and 217 and four epitopes between amino-acids Pol 346 and 387, using five different HLA-restricting elements. A new cluster of three conserved epitopes was found in the COOH segment of RT. CONCLUSIONS: This study shows that memory CTL responses against HIV-1 RT, integrase and protease are detectable in most patients at different stages of disease. The capacity of CTL to recognize simultaneously clusters of epitopes may become important for the immune control to reinforce antiretroviral drug efficiency. PMID- 9727565 TI - Mycobacterium tuberculosis infection and disease are not associated with protection against subsequent disseminated M. avium complex disease. AB - OBJECTIVE: To determine the relationship between Mycobacterium tuberculosis infection and disease and subsequent disseminated M. avium complex (MAC) disease in HIV-infected persons. DESIGN: A prospective observational cohort study. SETTING: The AIDS Linked to the Intravenous Experience (ALIVE) cohort of injecting drug users and the Johns Hopkins Hospital Adult HIV Clinic (JHHAHC). PARTICIPANTS: HIV-infected persons aged > 18 years with CD4 lymphocytes < 100 x 10(6)/l were followed between July 1989 and 31 October 1996. There were 182 persons in the ALIVE cohort and 1129 persons in JHHAHC who met these criteria. MAIN OUTCOME MEASURE: The relative risk of disseminated MAC was determined according to a history of prior opportunistic infection, MAC prophylaxis, antiretroviral therapy, M. tuberculosis infection or disease, race, sex, and injecting drug use. RESULTS: Amongst the 30 patients with active tuberculosis, eight developed disseminated MAC, compared with 208 cases of disseminated MAC amongst 1148 patients without prior M. tuberculosis infection or disease [relative risk (RR), 1.5; 95% confidence interval (CI), 0.8-2.7; P=0.2]. Amongst the 10 patients with extrapulmonary tuberculosis, five developed disseminated MAC (RR, 2.8; 95% CI, 1.5-5.2; P=0.02). Injecting drug use was associated with a decreased risk of disseminated MAC (RR, 0.7; 95% CI, 0.6-0.9; P=0.007). In a logistic regression analysis, disseminated MAC was significantly associated with extrapulmonary tuberculosis and other opportunistic disease, whereas antibiotic prophylaxis and injecting drug use were protective. CONCLUSIONS: A history of M. tuberculosis infection or disease was not associated with protection against subsequent disseminated MAC disease in HIV-infected persons. However, persons with extrapulmonary tuberculosis were at increased risk for disseminated MAC, particularly at low CD4 cell levels. PMID- 9727564 TI - Impairment of B-lymphocyte differentiation induced by dual triggering of the B cell antigen receptor and CD40 in advanced HIV-1-disease. AB - OBJECTIVE: This study was performed to investigate the hyporeactivity of purified B lymphocytes from HIV-1-infected patients. DESIGN: Given the importance of the B cell Ag receptor (BCR) and CD40 in B-lymphocyte activation, we assessed the capacity of purified peripheral blood B lymphocytes from HIV-1-infected patients to differentiate into Ig-secreting cells in a T-cell- and accessory-cell independent system of BCR and CD40 costimulation. METHODS: B lymphocytes from 21 HIV-1-infected patients were purified by immunomagnetic cell separation and costimulated with immobilized anti-CD40 monoclonal antibodies and Staphylococcus aureus Cowan I particles in the presence of interleukin (IL)-2 and IL-10. Homotypic aggregate formation, apoptosis, cell cycle entrance, proliferation and Ig secretion of B cells were analysed. RESULTS: Costimulation by the BCR and CD40 induced proliferation and differentiation of B lymphocytes into Ig-secreting cells in 13 patients (group I) but not in eight patients (group II). For three patients in group II, the dual triggering induced apoptosis of B cells. The unexpected inability of these cells to differentiate was associated with a high CD38 expression and a weak spontaneous production of Ig or anti-HIV-1 antibodies in patients with a high viral load and a low CD4+ lymphocyte count. Despite this anomaly, the B cells from group II were able to progress through the cell cycle after stimulation with a combination of phorbol ester and ionomycin in complete medium, suggesting an impairment in BCR and CD40 early signal transduction. CONCLUSION: Intrinsic in vitro hyporeactivity of B lymphocytes to dual triggering of BCR and CD40 was observed in advanced HIV-1 disease and appeared to be related to in vivo hyperactivation of B cells. PMID- 9727566 TI - Early regression of cervical lesions in HIV-seropositive women receiving highly active antiretroviral therapy. AB - OBJECTIVE: Advanced HIV disease is associated with a high prevalence of cervical squamous intra-epithelial lesions (SIL) and of infection with oncogenic human papillomavirus (HPV) genotypes. Triple-combination antiretroviral therapy results in decreased plasma HIV viral load, increased CD4 cell counts and partial restoration of immune functions in patients with severe HIV disease. This study investigated the outcome of SIL in HIV-seropositive women undergoing triple combination antiretroviral treatment. METHODS: Forty-nine women who started triple-combination antiretroviral therapy, including a protease inhibitor, were examined prior to and after a median 5-month treatment. We collected cytological, colposcopic and histologic data and assessed the presence of HPV DNA in cervical smears by PCR and Southern blot hybridization (SBH). RESULTS: The prevalence of SIL decreased from 69 to 53% during follow-up (P < 0.0001). Among 13 women who initially presented with high-grade SIL, conversion to lower grade was observed in two women and a full regression to normality was observed in one. Cytology also returned to normality in nine out of 21 women who initially presented with low-grade SIL. The high prevalence of HPV infection as detected by SBH and PCR was similar at the first and second examinations and the same high-risk viral genotypes were identified at both examinations in all infected patients but one. There was a higher increase in absolute CD4 cells in the subgroup of patients whose lesions regressed (99 versus 50 x 10(6)/l, P=0.03). CONCLUSION: Our observations demonstrate that active antiretroviral therapy may result in a reduced prevalence of cervical squamous intra-epithelial lesions despite the absence of clearance of HPV infection. PMID- 9727567 TI - Correlation of response to treatment and HIV genotypic changes during phase III trials with saquinavir and reverse transcriptase inhibitor combination therapy. AB - OBJECTIVES: Assessment of genotypic change in HIV protease during treatment with saquinavir (SQV) in combination with zidovudine (ZDV) and/or zalcitabine (ddC), to determine the influence of such changes on viral phenotype and response to treatment. DESIGN: Virologic substudies of Phase III clinical trials NV14256 and SV14604. METHODS: Population sequencing of HIV protease genes amplified from pre- and post-treatment plasma. Phenotyping of peripheral blood mononuclear cell (PBMC)-derived virus isolates, and genotyping of proviral DNA clones amplified from PBMC used in the expansion of virus isolates. RESULTS: In both trials the incidence of Met90 remained at < or = 20% in subjects receiving SQV in combination with ddC (with or without ZDV) for 1 year. A Val48 substitution was observed in two out of 81 subjects after 24 weeks and in two out of 75 subjects after 48 weeks. In 12 out of 13 NV14256 subjects with viral load rebound during SQV monotherapy these substitutions were associated with the rebound. In subjects treated with SQV plus ddC, rebound was associated with SQV resistance in six out of 22 cases and ddC resistance in five out of 22 cases. The incidences of non-BRU residues at positions 10, 63 and 71 were increased significantly (P < 0.05, Fisher's exact test) after SQV treatment with or without ZDV. However, comparison of genotypic and phenotypic data showed that these changes were not associated with reduced sensitivity to SQV. CONCLUSIONS: Virological failure during combination therapy can be due to resistance to either treatment drug, emphasising the need to change both the reverse transcriptase inhibitor and the protease inhibitor. Only Val48 and Met90 correlated directly with the development of reduced drug sensitivity during treatment with SQV in vivo. PMID- 9727568 TI - Kaposi's sarcoma in HIV infection: impact on opportunistic infections and survival. AB - OBJECTIVE: To determine the effect of Kaposi's sarcoma on survival of HIV infected patients. METHODS: Retrospective cohort study to compare the survival of 241 HIV-infected homosexual patients with Kaposi's sarcoma (cases) with that of 241 HIV-infected homosexual patients without Kaposi's sarcoma (control subjects) but with a similar level of immunosuppression (measured by the absolute CD4+ lymphocyte count). RESULTS: Cases and control subjects were similar in age, occurrence of previous opportunistic infections, and the use of antiretroviral therapy. The mean CD4+ lymphocyte counts were similar for cases and control subjects (185 x 10(6) versus 184 x 10(6)/l, respectively). Cases had a higher incidence of opportunistic infections (5.95 versus 3.88 infections, respectively, per 100 person-months of observation) and a greater number of infections typical of late-stage HIV infection. Cases had a shorter overall survival than did control subjects (P=0.0025). Kaposi's sarcoma was associated with an increased risk of death (odds ratio, 1.28), even when adjusting for age, previous opportunistic infection, baseline CD4+ lymphocyte count, and antiretroviral therapy. CONCLUSION: Kaposi's sarcoma appears to accelerate the clinical course of HIV infection. Opportunistic infections develop earlier and more often in patients with the disease than in control subjects. Survival was significantly shorter in patients with Kaposi's sarcoma. PMID- 9727569 TI - Rate of HIV-1 decline following antiretroviral therapy is related to viral load at baseline and drug regimen. AB - OBJECTIVES AND DESIGN: The dynamics uf viral decline following the initiation of antiretroviral treatment were studied in 29 HIV-1-infected patients participating in a two-arm trial comparing immediate (group A: ritonavir, zidovudine and lamivudine) and delayed (group B: ritonavir supplemented by zidovudine and lamivudine on day 21) triple therapy. Parameters underlying viral dynamics were estimated using mathematical models tailored to these treatment protocols. RESULTS: The decline in plasma HIV-1 density between day 0 and 21 was steeper in group A (-2.27+/- 0.46 log10) than group B (-1.87+/-0.56 log10). In a subset of patients amenable to full mathematical analysis, a short-lived productively infected cell compartment (producing approximately 97% of total virions) decayed with a half-life of 1.0-2.5 days, whereas a long-lived infected cell compartment decayed with a half-life of 18.8-32.8 days. Estimates for the time for the elimination of virus from these two cell populations ranged from 474 to 802 days. The rate of loss of productively infected CD4+ T cells was positively correlated with baseline viral load in group A and in the combined dataset. CONCLUSIONS: These results suggest that HIV-infected cell populations may have a faster turnover in patients with higher viral loads due to higher infection rate parameters, higher rates of virus production, or lower virus clearance rates. PMID- 9727570 TI - HIV combination therapy: partial immune restitution unmasking latent cryptococcal infection. AB - OBJECTIVE: To describe two cases of cryptococcal meningitis and one re exacerbation of Cryptococcus-associated meningitis occurring in temporal association with commencement of highly active antiretroviral therapy (HAART) in patients with advanced HIV infection (CD4 cells < 50 x 10(6)/l), which suggests that partial immune restitution can facilitate development of clinically apparent meningitis in response to Cryptococcus or its antigen. DESIGN: All HIV-infected patients with culture-proven cryptococcal meningitis diagnosed at a tertiary referral centre specialist infectious diseases unit from 1 January 1996 to 31 December 1996 were reviewed to examine the clinical and immunological parameters prior to and after commencing antiretroviral therapy. RESULTS: Three patients were diagnosed with clinically apparent meningitis within 7-39 days of changing or altering antiretroviral combination therapy consisting of zidovudine or stavudine, in combination with lamivudine and saquinavir. All patients had CD4 cell counts below 50 x 10(6)/l at initiation of therapy. Following institution of HAART, evidence of immune restitution was suggested by the following: (i) significant increases (3.7-14-fold) in numbers of CD4 cells (all three patients), (ii) significantly reduced (> 2-4 log10 reduction) HIV viral loads (two out of three patients), and (iii) prominent inflammatory changes in cerebrospinal fluid (white blood cells > 10 x 10(6)/l) at diagnosis (two out of three patients). CONCLUSIONS: Our report suggests that in patients with advanced HIV infection, partial immune restitution induced by HAART can precipitate onset of clinically apparent meningitis in those patients with latent cryptococcal central nervous system infection or with residual cryptococcal antigen present in the cerebrospinal fluid. PMID- 9727571 TI - A randomized trial of the effect of ritonavir in maintaining quality of life in advanced HIV disease. Advanced HIV Disease Ritonavir Study Group. AB - BACKGROUND: The aim of treatment for HIV disease is prolonging survival and improvement in health-related quality of life. Ritonavir is a potent, orally bioavailable HIV protease inhibitor with demonstrated impact on surrogate endpoints, AIDS-defining disease, and mortality. OBJECTIVES: To evaluate the effect of ritonavir combined with reverse transcriptase inhibitor therapy on patient functioning and well-being. METHODS: An international, multicenter randomized placebo-controlled clinical trial of ritonavir was conducted in HIV infected patients with CD4 cell counts < or = 100 x 10(6)/l. A total of 1090 patients were randomized to ritonavir and continued treatment with as many as two nucleoside agents (n=543) or placebo and continued treatment with as many as two nucleoside agents (n=547). Health-related quality of life was measured at baseline and after 3 and 6 months of treatment using the Medical Outcomes Study HIV Health Survey (MOS-HIV) and HIV-related symptoms scale. MOS-HIV contains 10 subscales and two summary scores (physical health and mental health). RESULTS: The two treatment groups were comparable on baseline CD4 cell counts, demographic characteristics, and MOS-HIV and HIV symptom subscale scores. After 3 months, statistically significant differences (P < 0.03) favoring the ritonavir-treated patients were seen on the physical health summary, mental health summary, and general health perceptions, social function, mental health, and energy/fatigue subscales. After 6 months of ritonavir therapy, significant differences were observed on physical health and mental health summary scores (P < 0.001), and on measures of general health perceptions, physical function, role function, social function, cognitive function, mental health, health distress, energy/fatigue, and overall ratings of quality of life (P < 0.01). Ritonavir-treated patients reported fewer fever symptoms and neurologic symptoms than the placebo group after 6 months of treatment (P < 0.005). CONCLUSIONS: Ritonavir therapy, combined with other antiretroviral treatments, significantly contributes to maintenance of functioning and well-being over at least 6 months in patients with advanced HIV disease. PMID- 9727572 TI - Preventing Mycobacterium avium complex in patients who are using protease inhibitors: a cost-effectiveness analysis. AB - BACKGROUND: Practice guidelines recommending Mycobacterium avium complex (MAC) prophylaxis for patients with HIV disease were based on clinical trials in which individuals did not receive protease inhibitors. OBJECTIVE: To estimate the cost effectiveness of strategies for MAC prophylaxis in patients whose treatment regimen includes protease inhibitors. DESIGN: Decision analysis with Markov modelling of the natural history of advanced HIV disease. Five strategies were evaluated: no prophylaxis, azithromycin, rifabutin, clarithromycin and a combination of azithromycin plus rifabutin. MAIN OUTCOME MEASURES: Survival, quality of life, quality-adjusted survival, health care costs and marginal cost effectiveness ratios. RESULTS: Compared with no prophylaxis, rifabutin increased life expectancy from 78 to 80 months, increased quality-adjusted life expectancy from 50 to 52 quality-adjusted months and increased health care costs from $233000 to $239800. Ignoring time discounting and quality of life, the cost effectiveness of rifabutin relative to no prophylaxis was $44300 per life year. Adjusting for time discounting and quality of life, the cost-effectiveness of rifabutin relative to no prophylaxis was $41500 per quality-adjusted life year (QALY). In comparison with rifabutin, azithromycin was associated with increased survival, increased costs and an incremental cost-effectiveness ratio of $54300 per QALY. In sensitivity analyses, prophylaxis remained economically attractive unless the lifetime chance of being diagnosed with MAC was less than 20%, the rate of CD4 count decline was less than 10 x 10(6) cells/l per year, or the CD4 count was greater than 50 x 10(6) cells/l. CONCLUSION: MAC prophylaxis increases quality-adjusted survival at a reasonable cost, even in patients using protease inhibitors. When not contraindicated, starting azithromycin or rifabutin when the patient's CD4 count is between 50 and 75 x 10(6) cells/l is the most cost effective strategy. The main determinants of cost-effectiveness are CD4 count, viral load, place of residence and patient preference. PMID- 9727573 TI - An open randomized controlled trial of zidovudine plus lamivudine versus stavudine plus lamivudine. AB - OBJECTIVE: To compare the antiretroviral effect and safety of zidovudine (ZDV) lamivudine (3TC) with that of stavudine (d4T)-3TC. METHODS: In an open randomized controlled trial antiretroviral therapy-naive patients who had CD4+ counts > or = 200 x 10(6)/l and plasma HIV RNA load > or = 10000 copies/ml were randomized to receive ZDV-3TC (200 mg three times daily and 150 mg twice daily, respectively) or d4T-3TC (40 mg and 150 mg, both twice daily). If the plasma HIV RNA level at week 8 or thereafter was > 500 copes/ml, indinavir was added at the next scheduled visit. Genotypic resistance analysis of the reverse transcriptase gene was performed at week 0 and 12. Results over 24 weeks were reported. RESULTS: Forty-seven patients were treated (24 took ZDV-3TC; 23 took d4T-3TC). Plasma HIV RNA levels decreased from median 4.80 to 3.15 log10 copies/ml (ZDV-3TC, P < 0.0001) and from 4.98 to 3.03 log10 copies/ml (d4T-3TC, P < 0.0001) after 12 weeks of treatment. Indinavir was added at week 12 in 11 out of 21 patients with ZDV-3TC and in 10 out of 22 patients with d4T-3TC. Median virus load at week 24 was 2.41 log10 and 2.29 log10 copies/ml (P=0.14), respectively. Seventy-five per cent (15 out of 20; ZDV-3TC) and 95% (18 out of 19; d4T-3TC) of patients had a virus load of < 500 copies/ml. Genomic evidence for 3TC resistance was found in all patients tested (11/11 ZDV-3TC and 12/12 d4T-3TC). At week 12, CD4 cell counts had increased with a median of 110 x 10(6)/l in the ZDV-3TC group (baseline, 315 x 10(6)/l) and a median of 115 x 10(6)/l in the d4T-3TC group (baseline 290 x 10(6)/l). At week 24, the median increases were 90 and 120 x 10(6)/l, respectively. Overall the increase of CD4+ cells was higher in the d4T 3TC group (P=0.02). CONCLUSION: d4T-3TC is at least as effective as ZDV-3TC, but 3TC resistance emerged in all patients investigated. The virological response of the dual nucleoside combination is of short duration. However, after addition of indinavir the virus load could be reduced to < 500 copies/ml in the majority of patients. The increase in CD4+ cell count was significantly greater in the d4T 3TC group. To prevent 3TC resistance, the drug should not be used in regimens containing only two nucleosides, irrespective the virus load at baseline. PMID- 9727575 TI - Social network dynamics and HIV transmission. AB - OBJECTIVE: To prospectively study changes in the social networks of persons at presumably high risk for HIV in a community with low prevalence and little endogenous transmission. METHODS: From a cohort of 595 persons at high risk (prostitutes, injecting drug users, and sexual partners of these persons) and nearly 6000 identified contacts, we examined the social networks of a subset of 96 persons who were interviewed once per year for 3 years. We assessed their network configuration, network stability, and changes in risk configuration and risk behavior using epidemiologic and social network analysis, and visualization techniques. RESULTS: Some significant decrease in personal risk-taking was documented during the course of the study, particularly with regard to needle sharing. The size and number of connected components (groups that are completely connected) declined. Microstructures (small subgroups of persons that interact intensely) were either not present, or declined appreciably during the period of observation. CONCLUSIONS: In this area of low prevalence, the lack of endogenous transmission of HIV may be related in part to the lack of a network structure that fosters active propagation, despite the continued presence of risky behaviors. Although the relative contribution of network structure and personal behavior cannot be ascertained from these data, the study suggests an important role for network configuration in the transmission dynamics of HIV. PMID- 9727574 TI - Effect of HIV-specific immune-based therapy in subjects infected with HIV-1 subtype E in Thailand. AB - OBJECTIVE: To examine the effect of treatment with an inactivated, gp120 depleted, HIV-1 immunogen (Remune) in 30 Thai subjects infected with HIV-1 subtype E. DESIGN: Sixty-week open-label study. METHODS: Thirty HIV-positive volunteers with CD4 cell counts > or = 300 x 10(6)/l were given intramuscular injections of Remune into the triceps muscle on day 1 and then at weeks 4, 8, 12, 24, 36, 48 and 60. RESULTS: Treatment with Remune was well-tolerated and augmented HIV-1-specific immune responses. Furthermore, subjects had a significant increase in CD4 cell count (P < 0.0001), CD4 cell percentage (P < 0.0001), CD8 cell percentage (P < 0.0001), and body weight (P < 0.0001) compared with pretreatment levels. Fourteen subjects with detectable viral load at day 1 showed a decrease at week 60 (P=0.04). Retrospective Western blot analysis showed 23 subjects with increased intensity of antibody bands and 15 patients showed development of new reactivities to HIV proteins, especially towards p17 and p15. CONCLUSION: These results indicate that HIV-specific immune-based therapeutic approaches such as Remune should be further examined in countries with different clades of HIV-1 and where access to antiviral drug therapies is limited. PMID- 9727576 TI - The incubation period to AIDS in injecting drug users estimated from prevalent cohort data, accounting for death prior to an AIDS diagnosis. AB - OBJECTIVE: To estimate the incubation-period distribution (time from seroconversion to AIDS) accounting for death before an AIDS diagnosis (DBAD) in a cohort of injecting drug users (IDU) in Amsterdam, The Netherlands and to compare these estimates with those previously obtained from a contemporaneous study of homosexual and bisexual men in Amsterdam carried out using the same facilities. DESIGN: Participants in a cohort study begun in Amsterdam at the end of 1985 have scheduled follow-up visits every 4 months. All participants of Dutch nationality and who had two or more follow-up visits before January 1996 from which CD4 measurements were available were included in this study. Data concerning AIDS diagnosis and death were verified through review of national and municipal registries. METHODS: Because time of seroconversion was unknown for study participants and because IDU are at substantial risk for DBAD, we used a Markov model with CD4-based stages that allows for DBAD. The parameters in this model were estimated using the method of maximum likelihood and confidence intervals were calculated using bootstrap methods. RESULTS: A total of 173 IDU (134 seroprevalent, 39 seroincident) made 1829 visits. Nearly 10% of the visits were non-consecutive. Forty-five IDU developed AIDS and 25 died without an AIDS diagnosis. We estimated that 24% [95% confidence interval (CI), 17-25%] of IDU die before an AIDS diagnosis. As a result, the median time from seroconversion to AIDS (10.5 years; 95% CI, 9.1-10.7 years) is considerably longer than the median time from seroconversion to death (8.3 years; 95% CI, 7.9-8.5 years). Conditional on survival to an AIDS diagnosis, the median time to AIDS is 8.2 years (95% CI, 7.7-8.7 years). The median survival time after a diagnosis of AIDS is estimated to be 1.0 years. CONCLUSION: The high occurrence of DBAD in IDU has a considerable influence on estimates of the incubation-period distribution. Progression from seroconversion to death was faster in the IDU cohort than in a cohort of homosexual men in Amsterdam (median, 8.3 years and 9.6 years, respectively). However, progression to AIDS conditional on survival to an AIDS diagnosis seems to be similar in both the IDU cohort and in the cohort of homosexual men (median, 8.2 years and 8.3 years, respectively). PMID- 9727577 TI - Sensitivity and specificity of a qualitative RNA detection assay to diagnose HIV infection in young infants. Perinatal AIDS Collaborative Transmission Study. AB - OBJECTIVE: To evaluate the sensitivity and specificity of an RNA detection assay for diagnosing perinatal HIV infection. METHODS: Plasma and serum specimens taken during the first 3 months of life from HIV-infected and uninfected children enrolled in a cohort study were assayed for HIV RNA using the qualitative nucleic acid sequence-based amplification (NASBA) kit. Sensitivity, specificity, and predictive values were calculated. NASBA results from infected children were compared with DNA PCR results from the same blood samples. Autoantibody patterns of suspected false-positive specimens were compared with those of subsequent specimens from the same child to exclude specimen labelling errors. RESULTS: Amongst 131 specimens from 105 HIV-infected children, the sensitivity of the qualitative NASBA assay was 13 out of 34 [38%; 95% confidence interval (CI), 22 56] at < 7 days, 56 out of 58 (97%; 95% CI, 88-100) at 7-41 days, and 37 out of 39 (95%; 95% CI, 83-99) at 42-93 days of life. Of 252 specimens from 206 uninfected children, six tested positive and one tested indeterminate by NASBA. Four of these positive specimens had discordant autoantibody patterns suggesting mislabelling; excluding these, the test specificity was 245 out of 248 (99%; 95% CI, 97-100). Amongst 128 paired specimens from infected children, NASBA results were more often positive than those from DNA PCR (103 versus 92; P=0.01). Amongst infants with specimens drawn in the first week of life, the proportion born after > 4 h of membrane rupture was greater amongst those testing negative (81%) than those testing positive (46%; P=0.05). CONCLUSIONS: The qualitative NASBA RNA assay is highly specific and more sensitive than DNA PCR. Qualitative RNA assays may be useful for diagnosing and excluding perinatal HIV infection in children after the first week of life for such purposes as initiating antiretroviral therapy and other treatment, resolving parental uncertainty, determining timing of transmission, and providing endpoints for intervention trials. PMID- 9727579 TI - Combination antiretroviral therapy for HIV infection: policies and practices. PMID- 9727578 TI - Taenia crassiceps cysticercosis and AIDS. PMID- 9727580 TI - Serum antibodies to dietary antigens in patients with HIV-1 infection. PMID- 9727581 TI - Once weekly azithromycin as prophylaxis against recurrence of non-tuberculous mycobacterial infections in HIV-1-positive individuals. PMID- 9727582 TI - Decline of anti-DP107 antibody associated with clinical progression. PMID- 9727583 TI - Partner notification in newly diagnosed HIV-positive patients. PMID- 9727584 TI - Who may be engaging in high-risk sex due to medical treatment advances? PMID- 9727585 TI - Four novel cytotoxic T-lymphocyte epitopes in the highly conserved major homology region of HIV-1 Gag, restricted through B*4402, B*1801, A*2601, B*70 (B*1509) PMID- 9727586 TI - HIV-1 subtypes in Santiago, Chile. PMID- 9727587 TI - HIV-1 group O virus infection in Abidjan, Cote d'Ivoire. PMID- 9727588 TI - The Y-chromosome and reproductive disorders. AB - Over the past decade the tools of modern molecular biology have provided unique insights into our fundamental understanding of developmental systems. These insights have been gleaned from the study of a wide variety of model organisms including yeast (Saccharomyces cerevisiae), fly (Drosophila), worm (Caenorhabditis elegans), and mouse. In man, the first analysis of developmental systems started with sexual differentiation and focused on the role of Y-linked genes. The presence of living developmental mutants in man affecting sexual development and the early technology of deletion mapping facilitated the isolation and identification of small segments of putative DNA suspected to contain sex-determining genes. The isolation of genes such as SRY (Sex Related gene on Y) has provided the first insights into the molecular biology of human sexual differentiation. The focus on the Y chromosome has brought further insights into chromosomal pairing, statural determinants in man, oncogenesis, spermatogenesis, haploid genomes, and the lineage of man himself. This paper provides the circumstantial and direct evidence to illustrate the importance of the Y chromosome in reproductive disorders, and in the analysis of haploid genomes. PMID- 9727589 TI - Epidemiological approaches to infertility. AB - There are a number of reasons why epidemiological approaches to infertility have not made a major contribution to research in Australia. They include the success of generic treatments, the high public profile of infertility and the consequent polarization of discussion over treatment versus prevention, some reluctance to draw attention to possible aetiologic factors which may be perceived negatively in public debates, and the lack of graduate training in reproductive and perinatal epidemiology. Voluntary infertility is now common for most of the fertile life span in developed countries, and intended family size is small. Many important conditions cannot be diagnosed without the use of invasive procedures or complex investigations, and the more widespread use of less invasive procedures has shown other conditions to be relatively common in healthy populations. If epidemiological approaches are to make a greater contribution towards an increased understanding and control of infertility, research should focus on retrospective and prospective cohort studies of the incidence and prevalence of infertility, nested case-control studies of occupational and environmental exposures, and an extension of the developing use of record-linkage across routinely collected data systems and registers. PMID- 9727590 TI - Genomic imprinting, development and disease--is pre-eclampsia caused by a maternally imprinted gene? AB - Several genes in conserved clusters are expressed from only the maternal or the paternal allele. The other allele has been genetically silenced ('imprinted') by its passage through one sex. Many known imprinted genes have effects on embryonic or trophoblast growth or fetal development, and mutation or loss of the single active copy causes diseases such as Prader-Willi, Angelmann and Beckwith Wiederman syndromes. Imprinted genes show an unusual mode of inheritance, since mutant genes have an effect on the phenotype only if they come from the parent from which they are expressed. This may explain some conditions which appear to be heritable but show an inconsistent pattern in affected families. Of particular interest is pre-eclampsia/eclampsia, the most serious complication of pregnancy, which has some features suggesting that it results from fetal expression of the mutant gene, but others which imply it results from maternal expression. This could be resolved by proposing that the condition is due to mutation in a paternally imprinted, maternally active gene which must be expressed by the fetus in order to establish a normal placenta in the first pregnancy. PMID- 9727591 TI - Properties and uses of embryonic stem cells: prospects for application to human biology and gene therapy. AB - Embryonic stem cells are pluripotent cells derived from the early mouse embryo that can be propagated stably in the undifferentiated state in vitro. They retain the ability to differentiate into all cell types found in an embryonic and adult mouse in vivo, and can be induced to differentiate into many cell types in vitro. Exploitation of ES cell technology for the creation of mice bearing predetermined genetic alterations has received widespread attention because of the sophistication that it brings to the study of gene function in mammals. Analysis of cell differentiation in vitro has also been of value, leading to the identification of novel bioactive factors and the elucidation of cell specification mechanisms. In this paper, we summarise the features of pluripotent cell lines and their applications, foreshadowing the impact that these systems may have on human biology. While the isolation of definitive human pluripotent cell lines has not yet been achieved, potential applications for these cells in the study of human biology, particularly cell specification, can be envisaged. Of particular interest is the possibility that human embryonic stem cells with properties similar to mouse embryonic stem cells might provide a generic system for gene therapy. PMID- 9727592 TI - Obesity: genes, glands or gluttony? AB - Distribution as well as amount of fat has health implications; central abdominal fat seems to be the major contributor to insulin resistance and risk of diabetes, hypertension and cardiovascular disease. Physical activity and diet affect overall adiposity; moreover, exercise specifically reduces visceral fat. The sexes differ in fat distribution; in particular, pre-menopausal women, despite greater overall adiposity, have much less visceral fat than men. There is a strong genetic determination of overall obesity and central abdominal adiposity. Genes regulating obesity (e.g. Ob) could modulate appetite, satiety, metabolic rate or physical activity. Moderate obesity probably results from interaction between genetic predisposition and an environment of abundant calories and reduced physical activity. Single gene mutations are being identified in a few morbidly obese people; however, the common genetic predisposition for obesity may relate to more subtle variations in regulatory controls. Diet and exercise are effective for some, but the response is often disappointing. Definition of pathways controlling appetite, metabolic rate and lipid metabolism may generate improved pharmacological compounds. Education and availability of lower-energy foods may help, but more radical approaches may be needed, such as environmental restructuring to increase physical activity. The problem is great, but failure will mean intolerably increased health costs. PMID- 9727594 TI - Disturbance of the reproductive axis induced by negative energy balance. AB - Animal reproduction is impaired when intake of energy is so restricted that activities essential to life are threatened; this is seen as a homeostatic adjustment that restricts wasteful energy expenditure. Fasting or exercising to a degree requiring considerable energy expenditure has major effects on the hypothalamus, including activation of corticotrophin-releasing factor (CRF) neurons, suppression of thyrotrophin-releasing hormone synthesis, and increased growth hormone secretion; these are associated with increased concentrations of hypothalamic neuropeptide Y mRNA and are corrected by administration of leptin, an adipose-tissue protein with a tertiary structure similar to the cytokine interleukin-2. This response to fasting results from a disordered pattern of activity in the gonadotrophin-releasing hormone (GnRH) pacemaker, characterized by reduced luteinizing hormone pulsatility, particularly during daytime. Animal studies have suggested that the response depends on an intact afferent vagal system from the stomach and the presence of oestrogen. Noradrenergic neurons forming the A2 group increase the activity of CRF neurons that, in turn, inhibit GnRH pulsatility. Reproductive impairment due to fasting is reversed by leptin, and abnormalities of leptin are described in individuals who fast or who develop exercise-induced amenorrhoea. This paper discusses these changes induced by negative energy balance and speculates on the involvement of leptin as a contributor to these abnormalities. PMID- 9727593 TI - Obesity and reproductive disorders: a review. AB - Obesity has significant consequences for the reproductive system, depending upon the amount and distribution of body fat. Epidemiological evidence clearly shows that being overweight contributes to menstrual disorders, infertility, miscarriage, poor pregnancy outcome, impaired fetal well-being and diabetes mellitus. Central adiposity is particularly important in clinical sequelae associated with an increased body mass index. The advent of assisted reproduction highlights the problems of being overweight, and the use of gonadotrophins in ovulation induction and in vitro fertilization is more difficult when the subject is overweight. Weight loss has marked effects on improving the menstrual cycle and promoting spontaneous ovulation and fertility. Results indicate that fertility is improved through exercise and sensible eating patterns when conducted in a group environment. The mechanisms for this are unclear but may be associated with changes in sensitivity to insulin. PMID- 9727595 TI - Biological bases of premature ovarian failure. AB - The ovary is endowed at birth with a fixed number of primordial follicles, which steadily dwindles throughout life as a result of atresia and recruitment towards ovulation. In addition to age, the number varies allometrically between species, larger and longer-lived animals tending to have more follicles initially and these disappear at a slower rate. A causal relationship between follicle depletion and menopause clearly exists, and there is a gradual acceleration of follicle wastage in the human ovary beginning more than a decade before the end of menstrual life. A mathematical model has provided confirmatory evidence of this relationship, and indicates that menopause is triggered by a threshold number of follicles which varies stochastically with a mean of 1100. PMID- 9727596 TI - Advances and challenges with intersex disorders. AB - Many recent advances have come from basic research into the mechanisms of sexual differentiation. These include the discovery of a number of genes (SRY, WT-1, SF 1, SOX-9) and one locus (DSS) involved in human gonadal differentiation, and understanding gained from extensive studies of the androgen receptor and its gene. The challenge for the clinician is to learn how best to use the new tools that have become available as a result of these advances. At the same time, community-based organizations whose members are adults with intersex disorders are issuing a challenge of a different kind: that the medical profession should radically revise its thinking on some of the most fundamental issues to do with treatment. The opportunity to take a fresh look at our policies in collaboration with well-informed patient groups is an advance as well as a challenge. Further research on both the origins and outcomes of intersex conditions promises to greatly improve the lives of affected people. PMID- 9727597 TI - Increased chromosome abnormalities in human preimplantation embryos after in vitro fertilization in patients with recurrent miscarriage. AB - Recurrent miscarriage is a pathological condition induced by maternal and embryonic causes. This paper describes a prospective study to determine the real incidence of aneuploidy for autosomes 13, 16, 18, 21, 22, and gonosomes in preimplantation human embryos obtained from patients with recurrent pregnancy loss after ovarian stimulation in an IVF-ET programme. Our results indicate that aneuploidy for the chromosomes analysed are abnormally higher in embryos obtained after IVF from recurrent abortion patients (58%) compared to non-recurrent abortion patients undergoing IVF. Furthermore, monosomies are six times more frequent than trisomies (47:8) in preimplantation embryos from recurrent abortion patients. Based on the present study, preimplantation genetic diagnosis (PGD) of embryos obtained from patients with recurrent miscarriage could prove advantageous in diagnosing abnormal embryos and selecting normal embryos for transfer. PMID- 9727598 TI - Are sperm counts really falling? AB - Semen quality is said to have been declining over the past 50 years. The biological significance of these changes is emphasized by a concomitant increase in the incidence of genitourinary abnormalities such as testicular cancer and cryptorchidism. The increase in regional frequency of testicular abnormalities over a relatively short period of time may be due to local environmental factors, including the comparatively recent fashion for wearing tight-fitting underwear. Data also indicates that prenatal exposure to environmental agents can affect the development of the male genital tract. From the reproductive point of view, an increased environmental impact on the human male gonad is of concern and merits the development of sensitive techniques that can detect deleterious agents. PMID- 9727599 TI - Genetic disorders and spermatogenesis. AB - Male infertility affects one man in twenty and a genetic basis seems likely in at least 30% of those men. Genetic regulation of fertility involves the inter related processes of testicular development, spermatogenesis (involving germ cell mitosis, meiosis and spermatid maturation), and their endocrine and paracrine regulation. In regard to spermatogenesis, particular attention has been given to the Yq11 region, where some spermatogenesis genes ('azoospermia factors') appear to be located. Several candidate genes have been identified but have not been shown to have a defined or essential role in spermatogenesis. Microdeletions of Yq11 are found in approximately 15% of azoospermic or severely oligospermic men. The complexity of the genetic control of male fertility is demonstrated by the evidence for genes involved in spermatogenesis and sexual differentiation on the X chromosome and autosomes. Better understanding of the genetic regulation of normal spermatogenesis will provide new probes for clinical studies; however, at present the majority of spermatogenic failure remains without an identified genetic linkage. The advent of intracytoplasmic sperm injection permits fertility in many previously sterile men and presents the possibility of their transmission of infertility; appropriate counselling is required. PMID- 9727600 TI - Antiandrogens as environmental endocrine disruptors. AB - Steroid hormone receptors control fundamental events in embryonic development and sex differentiation through their function as ligand-inducible transcription factors. The consequences of disrupting these processes can be especially profound during development due to the crucial role hormones play in controlling transient and irreversible developmental processes. Several environmental chemicals, including metabolites of the fungicide vinclozolin and the pesticide DDT, disrupt male reproductive development and function by inhibiting androgen receptor mediated events. A variety of in vitro and in vivo approaches have been used to determine the molecular basis of environmental antiandrogen toxicity. These chemicals commonly bind androgen receptor with moderate affinity and act as antagonists by inhibiting transcription of androgen dependent genes. PMID- 9727601 TI - Cystic fibrosis and reproduction. AB - Cystic fibrosis (CF) is the most common autosomal recessive disease. CF and congenital bilateral absence of the vas deferens (CBAVD) share a genetic and embryological background. Since the 1960s, medical therapy to reduce the progressive obstructive lung disease and nutritional deficiencies has resulted in most CF patients reaching adulthood. With the improved life expectancy of CF patients, new issues in reproductive health and pregnancy management have arisen. Puberty is delayed, with menarche often occurring eighteen months later than the average. Almost all men with CF are azoospermic. In both CF and CBAVD, the vas deferens is absent and the seminal vesicles are often hypoplastic. Many women with CF are subfertile, and if pregnancy is achieved there is an observed increase in maternal morbidity and mortality. The understanding of the molecular basis of CF and CBAVD has evolved, with the identification of hundreds of CF gene mutations and discovery of an associated intron polymorphism of the CF gene. The concept of severe and mild mutations has been introduced to explain the severe and mild phenotype variations such as the pancreatic insufficient and pancreatic sufficient patient. This paper reviews the above issues to assist with the management of infertile couples with CF or CBAVD. PMID- 9727602 TI - Potential benefits of cell cloning for human medicine. AB - The successful cloning of a mammal from an adult somatic cell nucleus opens new avenues for major advances in reproductive medicine, biotechnology and cellular based transplantation therapies for degenerative diseases. At the same time, this breakthrough has generated much heated discussion concerning the ethics of cloning. Twinning is a form of cloning, and there are instances in clinical assisted reproduction in which the deliberate formation of twins by embryo dissection would seem ethically acceptable. Nuclear transfer technology might facilitate the derivation of human embryonic stem cells, capable of differentiation into a wide variety of somatic cell lineages. Directed differentiation of human embryonic stem cells into specific cell types in vitro could provide a universal source of cells for transplantation therapy. The potential benefits of therapeutics based on cloning technologies are considerable, and hasty legislation to ban all such procedures could block progress in critical arenas of biomedical research. PMID- 9727603 TI - 9-Hydroxy-traumatin, a new metabolite of the lipoxygenase pathway. AB - 9-Hydroxy-traumatin, 9-hydroxy-12-oxo-10E-dodecenoic acid, was isolated as a product of 13S-hydroperoxy-9Z,11E-octadecadienoic acid as catalyzed by enzyme preparations of both soybean and alfalfa seedlings. This suggested that 9Z traumatin, 12-oxo-9Z-dodecenoic acid, was being converted into 9-hydroxy traumatin in an analogous manner to the previously identified enzymic conversion of 3Z-nonenal and 3Z-hexenal into 4-hydroxy-2E-nonenal and 4-hydroxy-2 E-hexenal, respectively. Other metabolites of 13S-hydroperoxy-9Z,11E-octadecadienoic acid were similar for both soybean and alfalfa seedling preparations, and they are briefly described. PMID- 9727604 TI - Rate constants for quenching singlet oxygen and activities for inhibiting lipid peroxidation of carotenoids and alpha-tocopherol in liposomes. AB - The (1)O2 quenching rate constants (kQ) of alpha-tocopherol (alpha-Toc) and carotenoids such as beta-carotene, astaxanthin, canthaxanthin, and lycopene in liposomes were determined in light of the localization of their active sites in membranes and the micropolarity of the membrane regions, and compared with those in ethanol solution. The activities of alpha-Toc and carotenoids in inhibiting (1)O2-dependent lipid peroxidation (reciprocal of the concentration required for 50% inhibition of lipid peroxidation: [IC50](-1)) were also measured in liposomes and ethanol solution and compared with their kQ values. The kQ and [IC50](-1) values were also compared in two photosensitizing systems containing Rose bengal (RB) and pyrenedodecanoic acid (PDA), respectively, which generate (1)O2 at different sites in membranes. The kQ values of alpha-Toc were 2.9 x 10(8) M(-1) s(-1) in ethanol solution and 1.4 x 10(7) M(-1) s(-1) (RB system) or 2.5 x 10(6) M(-1) s(-1) (PDA system) in liposomes. The relative [IC50](-1) value of alpha-Toc in liposomes was also five times higher in the RB system than in the PDA-system. In consideration of the local concentration of the OH-group of alpha-Toc in membranes, the kQ value of alpha-Toc in liposomes was recalculated as 3.3 x 10(6) M(-1) s(-1) in both the RB and PDA systems. The kQ values of all the carotenoids tested in two photosensitizing systems were almost the same. The kQ value of alpha-Toc in liposomes was 88 times less than in ethanol solution, but those of carotenoids in liposomes were 600-1200 times less than those in ethanol solution. The [IC50](-1) value of alpha-Toc in liposomes was 19 times less than that in ethanol solution, whereas those of carotenoids in liposomes were 60-170 times less those in ethanol solution. There were no great differences (less than twice) in the kQ and [IC50](-1) values of any carotenoids. The kQ values of all carotenoids were 40-80 times higher than that of alpha-Toc in ethanol solution but only six times higher that of alpha-Toc in liposomes. The [IC50](-1) values of carotenoid were also higher than that of alpha-Toc in ethanol solution than in liposomes, and these correlated well with the kQ values. PMID- 9727605 TI - Dietary oxidized cholesterol modulates cholesterol metabolism and linoleic acid desaturation in rats fed high-cholesterol diets. AB - The interactive effect of high dietary levels of oxidized cholesterol on exogenous cholesterol and linoleic acid metabolism was examined in male 4-wk-old Sprague-Dawley rats given high-cholesterol diets. The rats were pair-fed purified diets free of or containing either 0.5% cholesterol alone or both 0.5% cholesterol and 0.5% oxidized cholesterol mixture (containing 93% oxidized cholesterol) for 3 wk. Hepatic 3-hydroxy-3-methylglutaryl CoA reductase activity was reduced in rats given cholesterol alone or both cholesterol and oxidized cholesterol. However, hepatic cholesterol 7alpha-hydroxylase activity was lowered only when rats were given both cholesterol and oxidized cholesterol, although dietary cholesterol increased this activity. Reflecting this effect, acidic steroid excretion was lowest among the groups of rats given cholesterol and oxidized cholesterol. On the other hand, the activity of hepatic delta6 desaturase, a key enzyme in the metabolism of linoleic acid to arachidonic acid, was increased in rats given both cholesterol and oxidized cholesterol, although dietary cholesterol alone lowered its activity. As a result, the delta6 desaturation index, 20:3n-6 + 20:4n-6/18:2n-6, in liver and serum phospholipids tended to be higher in the group fed both cholesterol and oxidized cholesterol than in the one fed cholesterol alone. Thus, dietary oxidized cholesterol significantly modulated exogenous cholesterol metabolism and promoted linoleic acid desaturation even when it was given at high levels together with a high cholesterol diet. PMID- 9727606 TI - The cholesterol-lowering effect of guar gum in rats is not accompanied by an interruption of bile acid cycling. AB - A viscous hydrocolloid (guar gum, GG; 2.5% of the diet) or a steroid sequestrant (cholestyramine; 0.5% of the diet) was included in semipurified diets containing 0.2% cholesterol to compare the cholesterol-lowering effects of each agent in rats. In the present model, GG significantly lowered plasma cholesterol (-25%), especially in the density < 1.040 kg/L fraction, whereas cholestyramine was less potent. Bile acid fecal excretion significantly increased only in rats fed cholestyramine, similar to the cecal bile acid pool; the biliary bile acid secretion was accelerated by GG, but not their fecal excretion, whereas GG effectively enhanced neutral sterol excretion. As a result, the total steroid balance (+13 micromol/d in the control) was shifted toward negative values in rats fed the GG or cholestyramine diets (-27 or -50 micromol/d, respectively). Both agents induced liver 3-hydroxy-3-methylglutaryl-CoA reductase, but cholestyramine was more potent than GG in this respect. The present data suggest that, at a relative low dose in the diet, GG may be more effective than cholestyramine in lowering plasma cholesterol by impairing cholesterol absorption and by accelerating the small intestine/liver cycling of bile acids, which is interestingly, accompanied by reduction of bile acid concentration in the large intestine. PMID- 9727607 TI - Surface composition regulates clearance from plasma and triolein lipolysis of lipid emulsions. AB - Sphingomyelin (SM) and cholesterol (Chol) are major surface lipid constituents of plasma lipoproteins. We investigated the effects of SM and Chol on the plasma clearance of lipid emulsions as a model for lipoprotein particles in rats. The presence of Chol facilitated the removal of emulsion particles from plasma, whereas SM delayed particle removal. Preinjection of lactoferrin, an inhibitor of the apolipoprotein E (apoE) receptor, revealed that the differences in clearance of emulsions were due to the differences in affinity for the apoE receptor. Measurement of apolipoprotein binding suggested that the balance of apoE and apoC (apoC-II and apoC-III) bound to emulsions caused the difference in plasma clearance of emulsion particles. That is to say, SM in the emulsion surface decreased binding of apoE, which led to a longer circulation of emulsion particles in plasma. Chol, on the other hand, decreased the ratio of apoC to apoE, which may have promoted emulsion uptake through the apoE receptor. We also examined in vitro lipolysis using immobilized lipoprotein lipase (LPL) in a heparin affinity column. Lipolysis rates were significantly reduced by the incorporation of SM into the emulsion surface, but not by the incorporation of Chol, indicating that SM in the lipoprotein surface is an important lipid component regulating LPL-mediated lipolysis. Our results suggest that the presence of SM and Chol in the lipoprotein surface plays an important role in the circulation behavior and LPL-mediated lipolysis of lipid emulsions through their effect on the selectivity of plasma protein binding. PMID- 9727608 TI - Microsequencing of bovine cerebrospinal fluid apolipoproteins: identification of bovine apolipoprotein E. AB - In studies of bovine plasma lipoproteins, apolipoprotein E (apoE) was not found associated with alpha-lipoproteins isolated over a broad range of densities. However, studies of cerebrospinal fluid (CSF) lipoproteins from other mammals have shown that apoE is a major apolipoprotein associated with high density lipoprotein, a fact that prompted us to determine if this were also the case in bovine CSF. CSF samples were obtained from animals with a surgically implanted catheter. Most analyzed samples were obtained from cows at various stages of the postpartum period; however, a few samples also were obtained at term or during pregnancy. Analyses of isolated ultracentrifugal fractions by polyacrylamide gel electrophoresis revealed the presence of two apo, with the expected molecular weights for apoE and apoA-I. By using both matrix-assisted laser desorption mass spectrometry and microsequencing techniques, we demonstrated that these apo are indeed apoE and apoA-I. PMID- 9727609 TI - Remodeling of phospholipid fatty acids in mitochondrial membranes of estivating snails. AB - The effects of estivation on the phospholipid-specific fatty acid composition of mitochondrial membranes in the hepatopancreas of the terrestrial snail Cepaea nemoralis were investigated. The fatty acid composition of all phospholipids was significantly altered in snails estivating for 6 wk, indicating that substantial remodeling occurs. The most profound changes occurred in cardiolipin (CL). CL of estivating snails was 13-fold more saturated, contained 9-fold more monoenes, and had 45% fewer polyenes than in active snails. These differences were due, in part, to a reduction in linoleic acid (1 8:2n-6) content of CL from estivators. As in mammals, CL of active snails appears to preferentially incorporate 18:2n-6, which accounts for 60% of the acyl chains in this phospholipid. This proportion was reduced by 50% in estivators. Changes in the fatty acyl content of other phospholipids of estivating snails included increased monoenes in phosphatidylethanolamine (PE) and phosphatidylinositol, reduced ratios of n-3/n-6 polyenes in PE and phosphatidylcholine (PC), and an increased n-3/n-6 ratio in phosphatidylserine (PS). Arachidonic acid (20:4n-6) levels were reduced in PS but increased in CL and PC. Taken together, these alterations to fatty acid composition are consistent with decreased biological activity of membrane-related processes which occur in conjunction with the reduction of mitochondrial aerobic metabolism observed during estivation. PMID- 9727610 TI - Changes in tissue polyunsaturated fatty acids with age, in spontaneously hypertensive rats. AB - The relationship between the biosynthesis of long-chain fatty acids and their distribution in the key organs of hypertension is of considerable interest because of their role in the production of vasoactive eicosanoids and their effects on membrane properties. The present study analyzed the fatty acid compositions of the total lipids in the kidney, aorta, heart, and hepatocytes of 1-, 3-, and 6-mon-old spontaneously hypertensive rats (SHR) and their normotensive controls, Wistar Kyoto rats (WKY) by capillary gas chromatography . The major changes concerned the polyunsaturated fatty acids (PUFA). The percentage of arachidonic acid (AA) was significantly greater in the 1-mon-old SHR kidney than in the WKY kidney, but it was lower at 3 and 6 mon. The percentage of eicosapentaenoic acid was very low in the SHR kidney. The results for the aorta were similar, with marked decreases in 18:2n-6 and 18:3n-3 in SHR aged 1 and 6 mon. Despite a higher proportion of 18:2n-6 and AA at 6 mon, there was no major change in the SHR heart lipids. The fatty acid spectrum in the liver provides additional evidence for the previously reported inhibition of desaturase activities in SHR. Thus, this study shows that the PUFA composition is modified differently in different tissues in SHR, and this may be related to the pathogenesis of hypertension in these animals. PMID- 9727611 TI - A new conjugated linoleic acid isomer, 7 trans, 9 cis-octadecadienoic acid, in cow milk, cheese, beef and human milk and adipose tissue. AB - The identity of a previously unrecognized conjugated linoleic acid (CLA) isomer, 7 trans, 9 cis-octadecadienoic acid (18:2) was confirmed in milk, cheese, beef, human milk, and human adipose tissue. The 7 trans, 9 cis-18:2 isomer was resolved chromatographically as the methyl ester by silver ion-high-performance liquid chromatography (Ag+-HPLC); it eluted after the major 9 cis, 11 trans-18:2 isomer (rumenic acid) in the natural products analyzed. In the biological matrices investigated by Ag+-HPLC, the 7 trans, 9 cis-18:2 peak was generally due to the most abundant minor CLA isomer, ranging in concentration from 3 to 16% of total CLA. By gas chromatography (GC) with long polar capillary columns, the methyl ester of 7 trans, 9 cis-18:2 was shown to elute near the leading edge of the major 9 cis, 11 trans-18:2 peak, while the 4,4-dimethyloxazoline (DMOX) derivative permitted partial resolution of these two CLA isomers. The DMOX derivative of this new CLA isomer was analyzed by gas chromatography-electron ionization mass spectrometry (GC-EIMS). The double bond positions were at delta7 and delta9 as indicated by the characteristic mass spectral fragment ions at m/z 168, 180, 194, and 206, and their allylic cleavages at m/z 154 and 234. The cis/trans double-bond configuration was established by GC-direct deposition Fourier transform infrared as evidenced from the doublet at 988 and 949 cm(-1) and absorptions at 3020 and 3002 cm(-1). The 7 trans, 9 cis-18:2 configuration was established by GC-EIMS for the DMOX derivative of the natural products examined, and by comparison to a similar product obtained from treatment of a mixture of methyl 8-hydroxy- and 11-hydroxyoctadec-9 cis enoates with BF3 in methanol. PMID- 9727612 TI - Analysis of ubiquinone and tocopherol levels in normal and hyperlipidemic human plasma. AB - The type and amount of lipophilic antioxidants in plasma of hyperlipidemic patients are of great importance since they play a central role in preventing deleterious oxidation of blood lipids and proteins. Isolation and quantitation of lipophilic antioxidants from hyperlipidemic plasma samples meet great obstacles because of increased levels of various intermediary lipid products. This study was designed to develop a rapid and efficient extraction and separation procedure for simultaneous analysis of ubiquinone-9 and -10 as well as alpha-, delta-, and gamma-tocopherol isomers. The levels of ubiquinone-10, alpha- and gamma tocopherol were analyzed in human plasma samples using high-performance liquid chromatography. Lipid extraction was performed by petroleum ether/methanol/water. After phase separation, ubiquinone was reduced to ubiquinol by sodium borohydride and the lipids were separated on a C18 column. A binary gradient with solvents containing lithium perchlorate was used, and an electrochemical detector was employed for quantitation. This procedure was also efficient for the analysis of antioxidant lipids in samples containing a large number of accumulated and interfering lipid intermediates. Thus, the procedure described here is useful for efficient and rapid quantitation of ubiquinones and tocopherols in human plasma samples, especially those originating from hyperlipidemic patients. PMID- 9727613 TI - Evidence that commercial calf and horse sera can contain substantial amounts of trans-10,cis-12 conjugated linoleic acid. AB - We analyzed fetal calf, newborn calf, horse, and adult cow sera for conjugated linoleic acid (CLA). All sera samples contained CLA, but the amounts varied. The predominant isomer was cis-9,trans-11 CLA but some samples appeared to contain substantial amounts of an isomer with the retention time of trans-10,cis-12 CLA. PMID- 9727614 TI - Lectin may contribute to the atherogenicity of peanut oil. AB - Peanut oil is unexpectedly atherogenic for rats, rabbits, and primates. The lesions it produces are more fibrous than fatty. The mechanism underlying the atherogenicity of peanut oil has been elusive. Randomization of peanut oil reduces significantly its atherogenic properties, but native and randomized peanut oils have similar rates of lipolysis, and rats fed the two oils absorb and transport lipids in a similar fashion. Peanut oil differs from other oils in having a relatively high lectin content, and the randomization process markedly reduces the lectin content as well. The biologically active lectin of peanut oil has an affinity for glycoproteins found specifically on arterial smooth muscle cells. Peanut lectin has been shown to stimulate growth of smooth muscle and pulmonary arterial cells. Vigorous washing of peanut oil reduces its lectin content by 46%. Compared to rabbits fed cholesterol and peanut oil, rabbits fed cholesterol and washed peanut oil exhibited less severe atherosclerosis in the aortic arch (by 9%) and in the thoracic aorta (by 31%). The data suggest that peanut oils' endogenous lectin may contribute significantly to its atherogenic properties. PMID- 9727615 TI - Isolation and structure of glucosylceramides from the starfish Cosmasterias lurida. AB - From the water-insoluble lipid fraction of the methylene chloride/methanol extract of the starfish Cosmasterias lurida, two new glucosylceramides together with a known glucosylceramide, ophidiacerebroside E, were isolated by chromatographic procedures and characterized by spectroscopic (1H and 13C nuclear magnetic resonance, mass spectrometry) methods. The new compounds were identified as (2S,3R,4E,8E,10E)-1-(beta-D-glucopyranosyloxy)-3 -hydroxy-2-[(R)-2 hydroxyheptadecanoyl)amino]-9-methyl-4,8,10-o ctadecatriene (3) and (2S,3R,4E,8E,10E)-1-(beta-D-glucopyranosyloxy)-3 -hydroxy-2-[(R)-2 hydroxyoctadecanoyl)amino]-9-methyl-4,8,10-oc tadecatriene (4). PMID- 9727616 TI - The signature 10-hydroxy stearic acid thought to correlate with infectivity in oocysts of Cryptosporidium species is an artifact. AB - Heating or freezing leads to loss in infectivity of oocysts of Cryptosporidium parvum toward neonatal BALB/c mice and is reflected in the profile of the polar lipid fatty acids. Upon loss of infectivity, the ratio of polar lipid to neutral lipid fatty acid decreased and the relative proportions of 18:1n-9 also decreased; proportions of 18:2n-6 and 20:5n-6 increased, whereas the proportions of 16:0 remained constant with freezing. During these investigations, a novel fatty acid, 10-OH 18:0, was discovered in the glycolipid fraction. The identification of a fatty acid unique to species of Cryptosporidium was thought to provide a specific biomarker for this organism. Cryptosporidium also demonstrated fluctuations in absolute quantities of 10-OH 18:0 with events that lead to loss of infectivity. This led to the presumed correlation of this biomarker with infectious Cryptosporidium. The 10-OH 18:0 was putatively localized at the sn-2 position of phosphatidylethanolamine. High-performance liquid chromatography/electrospray ionization mass spectrometry revealed that the 10-OH 18:0 existed principally in the free fatty acid form. Herein, we establish that the free fatty acid 10-OH 18:0 was, in actuality, an artifact of the procedures for sample preparation. PMID- 9727617 TI - Rumenic acid: a proposed common name for the major conjugated linoleic acid isomer found in natural products. PMID- 9727618 TI - Genetic instability and chromosomal aberrations in colorectal cancer: a review of the current models. AB - Our understanding of the pathogenesis of cancer has undergone a revolution over the past decade. Tumors develop by the accumulation of damage to genes that regulate cell growth. Many of the genes responsible for disregulation of cell growth have been identified, as have the processes that lead to the genetic damage. One of the most important concepts that has facilitated our understanding of carcinogenesis is that of genetic or "genomic" instability, which is required to permit a sufficient amount of genetic damage to accumulate to permit the neoplastic phenotype to emerge and evolve. Two mechanisms that lead to genomic instability--one of which involves the loss of chromosomal fragments from the nucleus, and a second which is characterized by microsatellite instability--are discussed. PMID- 9727619 TI - Genetic abnormalities and microsatellite instability in colorectal cancer. AB - Our purpose was to investigate different genetic abnormalities, such as K-ras mutations, p53 alterations, and c-myc RNA overexpression, as well as microsatellite instability in 63 colorectal tumors obtained from patients that had undergone surgery. K-ras point mutations were analyzed by PCR-RFLP technique, followed by sequencing; p53 protein accumulation by immunohistochemistry; p53 gene mutations in exons 5-9 were studied by the SSCP and sequencing techniques, and c-myc overexpression by Northern blot. Microsatellite instability was performed at chromosomes 2p, 3p, and 11p by a PCR-based technique. Our data indicate a trend toward a poorer prognosis in patients who had K-ras transversions; besides, we have obtained a prevalence of c-myc RNA overexpression and p53 exon 7 mutations in the latest stages of tumor progression. In conclusion, our findings suggest that the recognition of molecular abnormalities might be used in colorectal cancer as a prognostic indicator or to determine the metastatic potential of colorectal adenocarcinomas. PMID- 9727620 TI - Epidemiology of sociodemographic characteristics, lifestyle, medical history, and colon cancer: a case-control study among French Canadians in Montreal. AB - Colon cancer is the second most common cancer in both men and women in North America and other developed countries. In a population-based case-control study of colon cancer among French Canadians in greater Montreal, a total of 402 cases and 668 controls were interviewed. The cancer cases were identified through the admission offices of five major Francophone teaching hospitals in Montreal from 1989 to 1993. The controls, matched by age, sex, place of residence, and language, were selected by a modified random digit dialing method. The results show that subjects who had ever been married had a lower risk for colon cancer (odds ratio [OR]: 0.58; 95% confidence interval [95% CI]: 0.48-0.84) than did individuals who had never been married. A significant association (OR: 1.90; p for trend = 0.003) was found between the height of subjects and the risk of colon cancer. The OR for individuals with a family history of colorectal cancer was 2.78 with a p value of 0.01. A direct and significant association (OR: 2.01) was found among constipation, use of laxatives (OR: 1.41), and the risk of colon cancer. Among women, a suggestive inverse association was detected between the number of full-term pregnancies and the risk of colon cancer in female subjects (the OR for five or more pregnancies was 0.58 with a p for trend of 0.08). There was also a suggestive linear trend (increased age-decreased risk) between age at menarche and the risk of colon cancer. No association was apparent between other sociodemographic characteristics and the risk of colon cancer. In conclusion, married individuals had lower risk for colon cancer, perhaps due to food habits or other characteristics of being single. Higher height and weight history 10 years before the diagnosis of cancer are risk factors for breast cancer, while both current weight and body mass index seem to be protective. Positive family history of colon cancer increased the risk of colon cancer significantly. PMID- 9727621 TI - Expression of lymphomagenic oncogenes in T-cell lymphomas of HPV 16 transgenic mice. AB - We have previously established that a dimer repeat of the complete HPV 16 genome is sufficient to cause multiple organ malignancies, either carcinomas or T-cell lymphomas, in transgenic mice. Here, we report the expression of oncogenes supporting the notion that these tumors arose via multiple oncogenic pathways. In these mice, the transgenic HPV 16 genome cosegregated with the tumor phenotype. E6/E7 expression was observed in both carcinomas and T-cell lymphomas, while E2 expression was observed only in T-cell lymphomas. Some of the T-cell lymphomas revealed E2 expression alone, implying that oncogenic pathways of HPV other than the one involving E6/E7 existed in these transgenic mice. To establish that this is the case, expression of genes downstream from E6/E7 and oncogenes involved in T-cell lymphoma formation were analyzed. p53 mutations were observed in two of five tumors that lacked E6 expression. High levels of c-myc gene expression were observed in five of six tumors with E7 expression, suggesting that a pathway involving E7, inactivation of Rb, and activation of c-myc is important in tumorigenesis of HPV 16 in these transgenic animals. High levels of expression of the c-Pim gene were also noted in two of three c-myc-expressing T-cell lymphomas, suggesting cooperation between these two proto-oncogenes. Activation of Hox-11, Tal2/SCL-2, and Rbtn1/Ttg1 expression, which are highly associated with human T cell acute lymphoblastic leukemia (T-ALL), was observed in three of three T-cell lymphomas with E2 expression but not E6/E7 expression, showing that pathways to tumor formation not involving E6/E7 exist in these transgenic animals. At least two oncogenic pathways to tumors in HPV 16 transgenic mice exist, one involving E6/E7 and c-myc and the other involving E2 and lymphomagenic oncogenes. PMID- 9727622 TI - Cytokeratin markers in malignant pleural mesothelioma. AB - Previously available serum tumor markers had a low clinical value in malignant pleural mesothelioma (MPM). The recently developed tissue polypeptide-specific antigen (TPS) and CYFRA 21-1 assays identify the soluble cytokeratin 18 and 19 fragments, respectively. In MPM these cytokeratins are expressed and may therefore be used as serum tumor markers. In this preliminary study, TPS and CYFRA 21-1 assays were evaluated to determine their potential for management of patients with MPM. Carcinoembryonic antigen (CEA) was evaluated as an additional marker. The study group consisted of 95 patients with benign lung and pleural diseases (BLPD), 14 patients with MPM, 41 patients with adenocarcinoma of lung (AC), and 40 patients with squamous cell carcinoma of lung (SQC). The utilized cutoff points corresponded to a 95% specificity for patients with BLPD. In MPM, TPS showed greater sensitivity (64.3%) than CYFRA 21-1 (50.0%), while CEA showed no sensitivity. In SQC, the marker CYFRA 21-1 had the highest sensitivity (52.5%), whereas in AC the most sensitive marker was CEA (56.1%). Significantly lower levels of CEA were found in MPM compared with BLPD (p < 0.001) or AC and SQC (p < 0.0001). Conversely, TPS levels in MPM were significantly higher than in SQC (p < 0.05). Close correlation of various individual pretreatment marker levels was observed only between TPS and CYFRA 21-1, both in MPM (r = 0.84; p < 0.001) and in non-small cell lung cancer (NSCLC) (r = 0.71; p < 0.001). In serial determinations of the markers during chemotherapy of MPM (n = 10), TPS and CYFRA 21-1 were shown to demonstrate more or less the same pattern of reactivity, although the changes in the TPS levels better reflected the clinical response to therapy. In conclusion, TPS seems to be a more sensitive marker than CYFRA 21-1. PMID- 9727623 TI - Predictive value of molecular alterations for the prognosis of urothelial carcinoma. AB - Accumulation of p53 and C-Myc overexpression are frequently found in advanced urothelial carcinomas. The prevalence and predictive value of both molecular alterations was investigated in 61 patients with superficial urothelial tumors. Distinct patterns of p53 accumulation and C-Myc overexpression were observed in superficial urothelial carcinoma of different stages. For instance, 67% of carcinomata in situ displayed accumulation of p53, but only 44% showed C-Myc overexpression, whereas in pT1 tumors the corresponding percentages were 25 and 75%. Similarly, while p53 accumulation was significantly (p = 0.02) associated with tumor grade, C-Myc overexpression did not correlate with grade. In multivariate analysis, p53 accumulation was found to be an independent predictor of tumor progression (p = 0.0096), whereas C-Myc overexpression did not correlate with the course of disease. Alterations in both markers together predicted neither tumor recurrence nor tumor progression better than p53 accumulation on its own. Sufficient expression of C-Myc may be a general requirement for proliferative competence in urothelial tumors, barring its use as a predictive marker. The predictive value of p53 accumulation for tumor progression was further underlined by the finding that in a distinct group of 52 patients with progressive urothelial carcinoma 73% of the recurrent tumors displayed p53 accumulation. PMID- 9727624 TI - Rapid increase in diagnosis of cutaneous melanoma in situ in Sweden, 1968-1992. AB - We analyzed incidence trends of cutaneous melanoma in situ in Sweden in 1968 1992. Among men, age-standardized rates increased from 0.1/100,000 in 1968 to 2.9/100,000 in 1992, which corresponds to an average annual increase of 15.0%. Among women, rates increased from 0.3 to 3.7, and the annual increase was 12.8%. Age-specific rates increased since 1978, predominantly in men aged 45 years and older, whereas in women rates increased in all ages. Multivariate analysis showed that incidence rates could be explained by both cohort effects and period effects in addition to age. However, cohort effects seemed more important among men than among women, in whom period effects dominated. Thus, factors associated with the development of melanoma in situ may differ between the sexes. Melanoma in situ is an immediate precursor of invasive melanoma, and the increased detection and surgical excision of these tumors will prevent the occurrence of invasive melanoma. PMID- 9727625 TI - Interstitial collagenase and the ED-B oncofetal domain of fibronectin are markers of angiogenesis in human skin tumors. AB - Collagenase-1 (C1) is the predominant matrix metalloproteinase present in newly formed microvessels and serves as a marker of neovascularization. The expression of the oncofetal fragment of fibronectin (Fn-f) was found to be increased during angiogenesis. In the present study, we investigated the relationship between the expression of collagenase-1 and the oncofetal fragment of fibronectin in newly formed microvessels as markers of tumor angiogenesis. In aggressive skin tumors (i.e., morpheaform and recurrent basal cell carcinomas) and squamous cell carcinomas, neovascularization was associated with a marked increase in the number of C1-positive and Fn-f-positive microvessels. At the beginning of elongation, microvessels begin to produce C1 but lose their ability to express type IV collagen and FVIII-related antigen. Later, this endothelium produces both Fn-f and C1. As maturation of microvessels occurs, C1-containing endothelium fails to express Fn-f but begins to produce a type IV collagen-containing basement membrane and FVIII-related antigen. These studies show that there is a selective expression of both Fn-f and collagenase by immature endothelial cells. C1 production begins at early stages of blood vessel formation and continues throughout angiogenesis. In contrast, Fn-f expression is limited to later stages of vasculogenesis, indicating that these proteins are reliable markers of angiogenesis. PMID- 9727627 TI - The role of protein glycosylation inhibitors in the prevention of metastasis and therapy of cancer. AB - Oligosaccharide moieties of cell-surface glycoproteins are thought to be involved in recognition events during cancer metastasis and invasion. Swainsonine, an inhibitor of the Golgi alpha-mannosidase II, has been shown to block pulmonary colonization by tumor cells and stimulate components of the immune system. Swainsonine also abrogates much of the toxicity of chemotherapeutic agents and stimulates bone marrow hematopoietic progenitor cells, suggesting additional therapeutic applications. We are currently characterizing the ability of swainsonine to modify cell growth in human and murine bone marrow progenitor cells. Furthermore, we are examining crucial steps in metastasis that depend upon cell surface molecules that play a role in cell-matrix interactions. Our work shows that tumor cell adhesion to collagen IV in vitro is rapidly stimulated by cis-polyunsaturated fatty acids and is dependent on protein kinase C activity. We are investigating the hypothesis that integrins are critical components of this adhesion and are examining potential signal transduction pathways that lead to the modulation of cell adhesion. PMID- 9727626 TI - Viral and nonviral gene delivery vectors for cancer gene therapy. AB - The development of vectors that are capable of efficient gene delivery is crucial to the success of gene therapy. We have developed both recombinant viral and nonviral vectors with the goal of correcting genetic abnormalities in cancer cells that are responsible for malignant transformation. Infection of cancer cells by recombinant adenovirus (Adv) indicates that the level of transduction is variable and dependent on the virus-to-cell ratio. Infection of cells with Adv/p53 resulted in levels of tumor suppressor p53 gene expression that could mediate tumor cell growth suppression and apoptosis, both in vitro and in vivo. The treatment of cancer cells with cisplatin prior to Adv transduction resulted in a higher level of therapeutic gene expression. Epidermal growth factor (EGF)/DNA complexes targeted to cancer cells overexpressing the EGF receptor resulted in efficient transduction of several lung cancer cell lines in vitro. As a result, these vectors provide improved methods with which to treat cancer in the clinical setting with gene therapy. PMID- 9727628 TI - Phorbol ester synergizes the dimethyl sulfoxide-dependent programmed cell death through diacylglycerol increment. AB - The regulation of cell proliferation or cell death by extracellular factors are the most intensely studied subjects in cell biology. Many conceptual problems remain to be clarified concerning the mechanisms that regulate the programmed cell death. In this work, we focus our attention on the possible role of protein kinase C activation during dimethyl sulfoxide (DMSO)-induced cell death. The present results suggest that the frequency of DMSO-dependent apoptosis of RPMI 8402 thymic lymphoma cells is increased by phorbol ester acetate supplementation. Enhancement of apoptosis can be abolished by cotreatment with the bisindolylmaleimide, a specific PKC inhibitor. The association between PMA and DMSO treatment provokes an early activation of an intracellular signaling mechanism that results, via sustained diacylglycerol elevation, in a possible long-term PKC activation. PMID- 9727629 TI - Nucleosome-releasing treatment makes surviving tumor cells better targets for nucleosome-specific anticancer antibodies. AB - Monoclonal antibody (MoAb) 2C5, a nucleosome-specific antinuclear autoantibody (ANA) from the repertoire of aged mice, was recently reported to recognize the surface of various tumor cells but not normal cells. Surface-bound nucleosomes (NSs) were previously proven to be MoAb 2C5's target on the outer membrane of tumor cells. Furthermore, MoAb 2C5 was found to have a strong antitumor effect during the early stages of tumor development. In an attempt to further increase antitumor effect of nucleosome-specific tumorocidal monoclonal antibody against established tumors, we investigated a possible way to enhance antibody association with tumor cells. Evidence is presented here demonstrating that the in vitro treatment of tumor cells (S49 T lymphoma) resulting in a partial cell death and massive liberation of intact NSs from dead tumor cells into the culture medium was accompanied by a 50-fold increase of MoAb 2C5 binding to the surface of surviving tumor cells. Massive NS release was observed in the case of S49 T cell treatment with dexamethasone and vincristine. However, a partial cell killing that was not accompanied with NS release (EL4 lymphoma treatment with doxorubicin) did not result in the enhanced binding of MoAb 2C5 to the surface of surviving tumor cells. The use of NS-specific tumorocidal antibodies, such as MoAb 2C5, in combination with another NS release-inducing tumor therapy, should provide an enhanced antibody-tumor binding. PMID- 9727631 TI - Alcohol effects on cytokine responses by immunocytes. PMID- 9727630 TI - Apoptosis and other mechanisms in androgen ablation treatment and androgen independent progression of prostate cancer: a review. AB - Patients with advanced prostate cancer commonly present with disseminated disease. For these patients, androgen ablation is a first-line treatment. This mode of therapy usually has an initially palliative effect on tumor-related symptoms and slows growth, although virtually all tumors eventually relapse to an androgen-independent, more aggressively growing phenotype. However, surprisingly little is known about the actions mediating the initial palliative effect as well as the initiation of androgen-independent tumor growth. In this review, some current concepts on mechanisms of androgen ablation treatment and androgen independent progression of prostate cancer is highlighted. Special attention is given to the involvement of apoptosis in these processes. PMID- 9727632 TI - Mechanism of HIV pathogenesis: role of superantigens in disease. AB - Initial infection with human immunodeficiency virus (HIV) results in a burst of viremia and an ensuing spread of virus to secondary lymphoid organs, after which a "latency" period occurs with little or no virus detectable in the circulation. The term latent period has been shown to be a misnomer, because substantial viral replication occurs during this time in lymph nodes, although clinically there appears to be few symptoms of disease. However, a telling indicator of active infection during this period is the initiation of decline in CD4+ T-cell numbers. A number of hypotheses have been postulated for the mechanism(s), as of yet not fully elucidated, by which T cells are depleted. Although quiescent cells can be infected, it has been shown that replication of HIV in CD4+ T cells requires cellular activation. The levels of viremia early in infection indicate that a large number of cells are actively infected, further suggesting that a mechanism must exist by which HIV activates a large pool of cells and ultimately causes their depletion. One possible mechanism for activation would be the presence of an HIV-encoded superantigen. Superantigens are proteins that are polyclonal stimulators of CD4+ T lymphocytes. This occurs as a result of their ability to form a trimolecular complex with MHC class II molecules on antigen-presenting cells and the Vbeta-specific region on the T-cell receptor. Thus, superantigen activation of T cells is antigen-nonspecific. The prototype superantigens are the staphylococcal enterotoxins. Putative viral superantigens include a protein from mouse mammary tumor virus and related retroviruses, rabies nucleocapsid, and the Nef protein of HIV. Nef is required for optimal HIV pathogenesis, and this may be due to its superantigen properties, where CD4 cells are transformed to the activated state for virus replication. PMID- 9727634 TI - Effect of chronic alcohol abuse on the CNS morbidity of HIV disease. PMID- 9727633 TI - Simian immunodeficiency virus, infection, alcohol, and host defense. PMID- 9727635 TI - Pneumococcal pneumonia in alcoholism and HIV infection. PMID- 9727636 TI - Immune reactivity to proteins biotransformed by alcohol metabolites. PMID- 9727637 TI - Alcohol and host defense against pulmonary infection with Pneumocystis carinii. PMID- 9727638 TI - Effects of ethanol consumption on mucosal and systemic T-cell-dependent immune responses to pathogenic microorganisms. PMID- 9727639 TI - Monocytes, alcohol use, and altered immunity. AB - The immunomodulatory capacity of acute, moderate alcohol consumption was investigated in this study in nonalcoholic volunteers after 2 ml of vodka/kg body weight of alcohol consumption. There was a significant, transient increase in interleukin-12 and interferon-gamma (IFNgamma) levels in whole blood samples collected 4 hr after alcohol consumption in response to an ex vivo bacterial challenge with lipopolysaccharide (p < 0.02). However, decreased IFNgamma levels were produced by mononuclear cells collected later after alcohol consumption (16 hr), suggesting that acute alcohol consumption has a biphasic effect on IFNgamma inducibility. Furthermore, isolated blood monocytes collected 16 hr after alcohol consumption showed significantly decreased IL-1beta production in response to subsequent bacterial stimulation, implying that in vivo alcohol consumption affects monocyte-derived inflammatory cytokine production. These results demonstrate that even acute, moderate alcohol consumption has a modulating capacity on immune functions that may contribute to decreased immunity and host defense. PMID- 9727640 TI - Alcohol consumption alters antigen-specific Th1 responses: mechanisms of deficit and repair. AB - Among the physiological effects associated with excessive alcohol consumption are alterations in immune function. Alcohol impairs T-helper 1 lymphocyte (Th1) regulated, cell-mediated immune responses. Antibody responses, regulated by T helper 2 lymphocyte (Th2), are either unimpaired or enhanced. Antigen presenting cells are central to the development of both Th1 and Th2 regulated immune responses. We used both T-cell receptor transgenic and conventionally immunized mice to demonstrate that ethanol consumption directly affects antigen presenting cells that, in turn, determines whether Th1 or Th2 response patterns predominate. Ethanol consumption inhibits Th1-associated interleukin-12 and interferon-gamma cytokine production and delayed-type hypersensitivity. Administration of exogenous recombinant interleukin-12 both restores interferon-gamma levels and delayed-type hypersensitivity responses in ethanol-consuming mice. PMID- 9727641 TI - The intracellular signaling network as a target for ethanol. PMID- 9727642 TI - Chronic ethanol consumption induces the production of tumor necrosis factor-alpha and related cytokines in liver and adipose tissue. AB - Increases in monocyte/macrophage production of the proinflammatory cytokine, tumor necrosis factor-alpha (TNF-alpha), parallel the evolution of liver injury in rats and humans with alcoholic liver disease. However, the possibility that TNF-alpha expression may be induced in other cell populations before serious liver disease develops has not been evaluated. To clarify this issue, mRNAs and/or protein levels of TNF-alpha and cytokines [interleukin (IL)-6, IL-10, transforming growth factor-beta (TGF)-beta, IL-12, and interferon-gamma] that regulate its biological activity were measured in sera, liver, and adipose tissues of rats that had developed hepatic steatosis after consuming ethanol containing diets for 6 weeks. Cytokine expression in the ethanol-fed groups was compared with that of pair-fed controls rats that had received isocaloric amounts of a similar, ethanol-free diet for the same time period. Animals were studied both before and after a surgical stress (partial hepatectomy) that is known to provoke cytokine production. Chronic ethanol consumption led to increased serum concentrations of TNF and related cytokines, at least in part, by inducing the overproduction of these factors in the liver and peripheral adipose tissues. Despite the pair-feeding protocol that ensured similar calorie consumption in both groups, adipose tissues in ethanol-fed rats also expressed more leptin, a TNF-alpha-inducible mRNA that encodes an appetite-suppressing hormone. Thus, white adipose tissue can be an important source of cytokines in nonobese animals and may be a target for ethanol's actions. These data implicate TNF-alpha as a potential mediator of the nutritional-metabolic aberrations that often accompany chronic alcohol intake, even in the absence of advanced liver disease. PMID- 9727643 TI - Ethanol alters the subcellular localization of cAMP-dependent protein kinase and protein kinase C. PMID- 9727644 TI - Mechanisms mediating the influence of alcohol on the hypothalamic-pituitary adrenal axis responses to immune and nonimmune signals. AB - In the rat, the acute administration of alcohol induces dose-related increases in plasma ACTH and corticosterone levels. This response depends on the delivery of the hypothalamic peptides corticotropin-releasing factor and vasopressin (VP) to the pituitary. On the other hand, exposure to an alcohol diet for 7 to 10 days significantly blunts the hypothalamic-pituitary-adrenal (HPA) axis in response to other homeostatic threats, such as mild electroshocks or immune signals. This decreased response is at least in part due to an attenuated ability of VP to increase ACTH secretion. We have previously shown that nitric oxide (NO) inhibits the pituitary response to VP. We therefore hypothesized that chronic alcohol treatment might increase levels of this gas within the HPA axis, and tested this possibility by determining whether blockade of NO formation might restore a normal pituitary response to VP. We observed that such was the case, and therefore propose that NO participates in the blunted activity of the HPA axis during prolonged exposure to alcohol. Finally, we determined whether alcohol would exert neuroendocrine effects that extended beyond the initial drug treatment. To explore this possibility, we injected rats with alcohol intragastrically for 3 days, then re-exposed them to the drug 3 to 12 days later. Rats pretreated with the vehicle and injected with alcohol several days later showed the expected significant rise in plasma ACTH and corticosterone levels, as well as a marked increase in immediate early genes mRNA levels in the paraventricular nucleus (PVN) of their hypothalamus. In contrast, animals pretreated with alcohol exhibited a blunted hormonal and hypothalamic response during the second drug exposure but, interestingly, retained a normal endocrine response to other signals, such as exposure to electro-footshocks or cytokine injection. We originally thought that this phenomenon of selective endocrine tolerance might be explained by decreased serotonin levels in the PVN. Whereas alcohol indeed decreased concentrations of this neurotransmitter in the PVN, exposure to electroshocks induced similar changes. However, an initial exposure to shocks did not blunt the ability of the HPA axis to be activated by a second shock session, by alcohol or by immune signals (delivered several days later). These results do not support the hypothesis that the decreased HPA axis response to a second alcohol challenge is mediated via decreased serotonin afferents to corticotropin-releasing factor neurons in the PVN. PMID- 9727645 TI - Tumor necrosis factor and alcoholic liver disease. AB - Increased levels of hepatic and serum tumor necrosis factor (TNF) have been documented in animal models of alcoholic liver disease and in human alcoholic liver disease. This dysregulated TNF metabolism has been postulated to play a role in many of the metabolic complications and the liver injury of alcoholic liver disease. One potential therapy for alcoholic liver disease may be agents that downregulate TNF production or block TNF activity. Indeed, agents such as prostaglandins and glucocorticoids (both inhibit TNF production) have been used in both human liver disease and experimental models of liver injury, and anti-TNF antibody has recently been shown to attenuate the hepatotoxicity in an animal model of alcoholic-related liver disease. In this study, we demonstrate that a simple ex vivo system can be used to initially assess potential efficacy of anticytokine agents when administered to humans. Both prednisone and a prostaglandin analog were effective in downregulating TNF and interleukin-8 production. The liver is normally resistant to TNF cytotoxicity. Sensitivity to TNF cytotoxicity is thought to occur when there is inadequate production of hepatic protective factors. In this study, we showed that, when patients with acute alcoholic hepatitis were matched with trauma patients for serum levels of interleukin-6, they had similar depressions in the negative acute phase protein, albumin, but markedly different increases in the major acute phase protein, C reactive protein. Patients with alcoholic hepatitis had a very blunted response. We also showed that inhibiting activation of the redox sensitive transcription factor NFkappaB sensitizes to TNF-induced hepatocyte death in vitro. This transcription factor is important for the production of both cytokines and many acute phase protective factors. Several hepatic protective factors are induced by TNF. One possible mechanism for liver injury in alcoholic hepatitis may be inadequate generation of hepatic protective factors. Our future understanding of mechanisms of alcoholic liver disease will involve understanding the balance between noxious and protective factors in the liver, and this should lead to rational therapy for this disease process. PMID- 9727646 TI - Molecular mechanism of induced hepatic macrophage cytokine gene expression in experimental alcoholic liver injury. PMID- 9727647 TI - The role of Kupffer cells and reactive oxygen species in hepatic injury during acute and chronic alcohol intoxication. PMID- 9727648 TI - The potential mechanism of induction of inducible nitric oxide synthase mRNA in alveolar macrophages by lipopolysaccharide and its suppression by ethanol, in vivo. AB - This study reviews the putative mechanism of ethanol (ETOH)-mediated downregulation of inducible nitric oxide synthase (iNOS) messenger RNA (mRNA) and protein and upregulation of constitutive NOS activity (ecNOS) in immunocompetent cells and endothelium, in vivo. Current evidence supports the hypothesis that ETOH inhibits the phospholipase D-tyrosine kinase pathway involved in the phosphorylation and activation of NADPH oxidase and myeloperoxidase, which upregulates the formation of reactive oxygen intermediates and mitogen-activated protein kinase cascade, including the extracellular receptor-linked kinase 1 and 2 (erk1 and erk2). This decreases reactive oxygen intermediate formation, tyrosine kinase-induced phosphorylation, and activation of transcription factors that, in turn, decreases the expression of iNOS mRNA. Also, ETOH-mediated attenuation of endotoxin-induced downregulation of nuclear protein kinase C activity appears to decrease the stability of expressed iNOS mRNA. ETOH-mediated inhibition of tyrosine kinase activity may also explain the ability of ETOH to upregulate ecNOS enzymatic activity, because tyrosine kinase activity suppresses ecNOS enzymatic activity. PMID- 9727649 TI - Transgenic mouse model of ethanol as a cofactor in HIV disease. PMID- 9727650 TI - Cefprozil and respiratory tract infections in children. AB - Respiratory tract infections are the leading cause of children's visits to physicians. Antibiotic resistance is increasing among the three most common bacterial respiratory pathogens, Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis, which cause acute otitis media and sinusitis. Therefore clinicians may need to reexamine their treatment strategies because these infections may cause complications. Antibiotic selection in such infections should take into account local bacterial resistance patterns, anticipated clinical efficacy against S. pneumoniae, H. influenzae and M. catarrhalis, including penicillin-resistant and beta-lactamase-producing strains, as well as the safety profile and compliance-enhancing features of the drug. PMID- 9727651 TI - Microbiology of bacterial respiratory infections. AB - The upper respiratory tract may become susceptible to bacterial infection as a result of health conditions such as allergies and viral infections, as well as the effects of smoking and airborne environmental pollutants. Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis are the most common bacterial pathogens in upper and lower respiratory tract infections. Streptococcus pyogenes is the predominant bacterial pathogen in pharyngitis and tonsillitis. Bacterial pathogens adhere to mucous membranes and colonization ensues. In an otherwise healthy individual the host immune system responds to the invading bacteria resulting in edema and swelling. If antimicrobial treatment does not eradicate the invading organisms and successfully interrupt the progress of the infection, the patient may develop recurrent or chronic disease. S. pneumoniae and other pathogens once susceptible to penicillin and other antibiotics are now becoming resistant. Bacterial resistance has developed and disseminated because of the widespread use of antibiotics. Major mechanisms of bacterial resistance to antimicrobials in upper respiratory tract infections include enzymatic inhibition, membrane impermeability, alteration of target enzymes, active pumping out of antibiotic and alteration of the ribosomal target. PMID- 9727652 TI - Microbial factors leading to recurrent upper respiratory tract infections. AB - The treatment or prophylaxis of upper respiratory tract infections such as otitis media, sinusitis and tonsillitis with penicillins can generate bacterial resistance caused by production of beta-lactamase or changes in the penicillin binding proteins. This resistance can spread in the community even to untreated individuals. The prevalence of resistant organisms tends to increase in the winter months. Beta-lactamase-producing bacteria may interfere with the eradication of penicillin-susceptible organisms and may account for substantial numbers of therapeutic failures among cases of otitis media, sinusitis and tonsillitis. The presence of normal flora that possess interfering capabilities against potential pathogens is beneficial to the host. Such flora may enhance recovery and prevent infections of the tonsils by group A beta-hemolytic streptococci. Therapeutic use of antimicrobial agents that preserve the normal flora but overcome penicillin-susceptible or -resistant pathogens may enhance recovery from upper respiratory tract infections. PMID- 9727653 TI - Clinical perspectives on sinusitis and otitis media. AB - As the leading cause of physician office visits and loss of time from school as well as the cause of significant morbidity among young children, respiratory infections impose a major burden on the health care system. The most common causative pathogens are Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis. In young children acute otitis media and sinusitis may present with relatively nonspecific symptoms such as irritability. Older children may complain of more specific problems; for example those with otitis media may complain of otalgia. Upper respiratory tract infections are typically diagnosed by signs and symptoms and treated empirically with an antimicrobial agent that offers coverage of the usual causative respiratory pathogens. PMID- 9727654 TI - Upper respiratory tract infections in family practice. AB - The causative pathogens in the majority of mild to moderate upper respiratory tract infections are Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis. Bacterial infections of the respiratory tract are often treated empirically; however, the recent increase in serious infections caused by S. pneumoniae and rising antimicrobial resistance rates have prompted experts to reevaluate the therapeutic approaches to treatment of these infections. Although amoxicillin continues to be considered a first line therapy, some situations warrant alternative therapies. Antimicrobial therapy must provide effective coverage of the potential pathogens, yet issues of compliance must also be addressed to ensure clinical success. Ease of administration, taste and the potential for adverse events are important considerations for the pediatric population. Clinical trials support the use of alternative therapies in the treatment of patients with upper respiratory tract infections. PMID- 9727655 TI - Potential infectious disease complications of upper respiratory tract infections. AB - Upper respiratory tract infections (URTIs), particularly otitis media and sinusitis, are prevalent among children. With recurrent URTIs there is an increased likelihood of sequelae. Suppurative complications associated with URTIs, although rare, must be treated rapidly to prevent serious morbidity and mortality. Further, increase in antimicrobial resistance may be accompanied by an increased risk for complications because infecting pathogens may be more difficult to eradicate. PMID- 9727656 TI - Treatment of upper and lower respiratory tract infections: clinical trials with cefprozil. AB - The oral second generation cephalosporin cefprozil has a broad spectrum microbiologic profile, with good in vitro activity against respiratory pathogens; 90% or more of Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis isolates are susceptible to cefprozil. Clinical trials of cefprozil have consistently demonstrated good clinical success rates in upper and lower respiratory tract infections, including otitis media, sinusitis, pharyngitis/ tonsillitis and acute bacterial exacerbations of chronic bronchitis. Most recently cefprozil has demonstrated success in children with recurrent and persistent acute otitis media. Data from clinical trials including more than 4000 children and adults have shown that cefprozil is well-tolerated. The most common adverse events associated with cefprozil are gastrointestinal disturbances (i.e. diarrhea and nausea). In two patient satisfaction surveys (pediatric and adult), cefprozil was cited for having a low incidence of side effects and was rated by children as having a pleasing taste. These data indicate that cefprozil is a practical therapeutic choice for the treatment of upper and lower respiratory tract infections. PMID- 9727657 TI - Effect of ventricular shock strength on cardiac hemodynamics. AB - INTRODUCTION: The effect of implantable defibrillator shocks on cardiac hemodynamics is poorly understood. The purpose of this study was to test the hypothesis that ventricular defibrillator shocks adversely effect cardiac hemodynamics. METHODS AND RESULTS: The cardiac index was determined by calculating the mitral valve inflow with transesophogeal Doppler during nonthoracotomy defibrillator implantation in 17 patients. The cardiac index was determined before, and immediately, 1 minute, 2 minutes, and 4 minutes after shocks were delivered during defibrillation energy requirement testing with 27- to 34-, 15-, 10-, 5-, 3-, or 1-J shocks. The cardiac index was also measured at the same time points after 27- to 34-, and 1-J shocks delivered during the baseline rhythm. The cardiac index decreased from 2.30 +/- 0.40 L/min per m2 before a 27- to 34-J shock during defibrillation energy requirement testing to 2.14 +/- 0.45 L/min per m2 immediately afterwards (P = 0.001). This effect persisted for > 4 minutes. An adverse hemodynamic effect of similar magnitude occurred after 15 J (P = 0.003) and 10-J shocks (P = 0.01), but dissipated after 4 minutes and within 2 minutes, respectively. There was a significant correlation between shock strength and the percent change in cardiac index (r = 0.3, P = 0.03). The cardiac index decreased 14% after a 27- to 34-J shock during the baseline rhythm (P < 0.0001). This effect persisted for < 4 minutes. A 1-J shock during the baseline rhythm did not effect the cardiac index. CONCLUSION: Defibrillator shocks > 9 J delivered during the baseline rhythm or during defibrillation energy requirement testing result in a 10% to 15% reduction in cardiac index, whereas smaller energy shocks do not affect cardiac hemodynamics. The duration and extent of the adverse effect are proportional to the shock strength. Shock strength, and not ventricular fibrillation, appears to be most responsible for this effect. Therefore, the detrimental hemodynamic effects of high-energy shocks may be avoided when low-energy defibrillation is used. PMID- 9727658 TI - Shortening of the intraventricular conduction time in premature ventricular beats during ventricular extrastimulation: possible role of dimensional shortening of the conducting distance during ventricular systole. AB - INTRODUCTION: Conduction time (CT) is given by the formula: conducting distance divided by conduction velocity. Based on this formula, we hypothesized that CT shortening (i.e., supernormal conduction) may result from dimensional shortening of the distance of impulse propagation, which naturally occurs during ventricular systole. METHODS AND RESULTS: To test the above, two separate groups of patients were studied, group A (14 patients) for electrophysiologic study and group B (12 patients) for echocardiographic study. In group A patients, CT from the stimulus artifact to the basal lateral wall of the left ventricle (LV) (S-LV interval) was measured using right ventricular (RV) apical extrastimulus testing. S-LV interval shortening in premature RV beats was demonstrated in all 14 patients. The maximum shortening was 20 +/- 9 msec (range 10 to 40), and the maximum % shortening was 16% +/- 6% (7% to 27%). In group B patients with implanted pacemakers, the major (long) and minor (short) axis dimensions of the LV were measured with echocardiography. The major axis dimension was used as an approximate measure of the linear length from the RV apex to the basal lateral wall of LV. The maximum % shortening of the major axis dimensions was 15% +/- 4%, 16% +/- 2%, and 11% +/- 4% during VVI pacing, respectively, at paced cycle lengths of 1,000 (11 patients), 800 (5 patients), and 600 msec (12 patients). The maximum % shortening of the S-LV intervals was comparable in magnitude with that of the major axis dimensions: 20% versus 15% +/- 4%, 15% +/- 7% versus 16% +/- 2% and 16% +/- 6% versus 11% +/- 4%, respectively, at paced cycle lengths of 1,000, 800, and 600 msec. There was also a good temporal correlation between the electrophysiologic (CT shortening) versus echocardiographic (dimensional shortening) parameters. Thus, the intraventricular CT and the major axis dimension of the LV were shortened in a similar magnitude and also at a similar timing in the cardiac cycle. CONCLUSION: These findings suggest the possibility that supernormal conduction may result, at least in part, from dimensional shortening of the pathway length of impulse propagation from the stimulating to recording electrodes, which naturally occurs during ventricular systole. PMID- 9727660 TI - Differential effect of adenosine on anterograde and retrograde fast pathway conduction in patients with atrioventricular nodal reentrant tachycardia. AB - INTRODUCTION: Several studies have shown that the fast pathway is more responsive to adenosine than the slow pathway in patients with AV nodal reentrant tachycardia. Little information is available regarding the effect of adenosine on anterograde and retrograde fast pathway conduction. METHODS AND RESULTS: The effects of adenosine on anterograde and retrograde fast pathway conduction were evaluated in 116 patients (mean age 47 +/- 16 years) with typical AV nodal reentrant tachycardia. Each patient received 12 mg of adenosine during ventricular pacing at a cycle length 20 msec longer than the fast pathway VA block cycle length and during sinus rhythm or atrial pacing at 20 msec longer than the fast pathway AV block cycle length. Anterograde block occurred in 98% of patients compared with retrograde fast pathway block in 62% of patients (P < 0.001). Unresponsiveness of the retrograde fast pathway to adenosine was associated with a shorter AV block cycle length (374 +/- 78 vs 333 +/- 74 msec, P < 0.01), a shorter VA block cycle length (383 +/- 121 vs 307 +/- 49 msec, P < 0.001), and a shorter VA interval during tachycardia (53 +/- 23 vs 41 +/- 17 msec, P < 0.01). CONCLUSION: Although anterograde fast pathway conduction is almost always blocked by 12 mg of adenosine, retrograde fast pathway conduction is not blocked by adenosine in 38% of patients with typical AV nodal reentrant tachycardia. This indicates that the anterograde and retrograde fast pathways may be anatomically and/or functionally distinct. Unresponsiveness of VA conduction to adenosine is not a reliable indicator of an accessory pathway. PMID- 9727659 TI - Conduction properties of the crista terminalis in patients with typical atrial flutter: basis for a line of block in the reentrant circuit. AB - INTRODUCTION: Previous mapping studies in patients with typical atrial flutter have demonstrated the crista terminalis to be a posterior barrier of the reentrant circuit forming a line of block. However, the functional role of the crista terminalis in patients with or without a history of atrial flutter is not well known. The aim of this study was to determine whether the conduction properties of the crista terminalis are different between patients with and those without a history of atrial flutter. METHODS AND RESULTS: The study population consisted of 12 patients with clinically documented atrial flutter (group 1) and 12 patients with paroxysmal supraventricular tachycardia as well as induced atrial flutter (group 2). A 7-French, 20-pole, deflectable Halo catheter was positioned around the tricuspid annulus. A 7-French, 20-pole Crista catheter was placed along the crista terminalis identified by the recording of double potentials with opposite activation sequences during typical atrial flutter. After sinus rhythm was restored, pacing from the low posterior right atrium near the crista terminalis was performed at multiple cycle length to 2:1 atrial capture. No double potentials were recorded along the crista terminalis during sinus rhythm in both groups. In group 1, the longest pacing cycle length that resulted in a line of block with double potentials along the crista terminalis was 638 +/- 119 msec. After infusion of propranolol, it was prolonged to 832 +/- 93 msec without change of the interdeflection intervals of double potentials. In group 2, the longest pacing cycle length that resulted in a line of block with double potentials along the crista terminalis was 214 +/- 23 msec. After infusion of procainamide, it was prolonged to 306 +/- 36 msec with increase of interdeflection interval of double potentials. CONCLUSION: The crista terminalis forms a line of transverse conduction block during typical atrial flutter. Poor transverse conduction property in the crista terminalis may be the requisite substrate for clinical occurrence of typical atrial flutter. PMID- 9727661 TI - Delayed activation of cardiac swelling-induced chloride current after step changes in cell size. AB - INTRODUCTION: A lag phase has been reported for the activation of cardiac swelling-induced chloride currents. Prior demonstrations of this lag used methods that produce gradual changes in cell size, making interpretation and quantification of the time course problematic. METHODS AND RESULTS: Isolated dog atrial cells were studied using the whole cell, patch clamp technique. Step changes in cell size were produced by application of transient pulses of positive pressure, and the time course for activation of the swelling-induced chloride current was observed. There was a distinct temporal dissociation between size changes and current activation that was temperature sensitive. Activation half times were 98 +/- 31 seconds and 586 +/- 112 seconds at 36 degrees C and room temperature, respectively. Swelling-induced chloride currents were evoked in a higher percentage of cells at 36 degrees C (83%) compared with room temperature (50%). CONCLUSION: Cardiac swelling-induced chloride current activates with a distinct lag after step changes in cell size. The activation time course is temperature sensitive. These observations are consistent with the notion that signal transduction events, and not simply membrane stretch, are required for the activation of cardiac swelling-induced chloride current. PMID- 9727662 TI - Ability of activation recovery intervals to assess action potential duration during acute no-flow ischemia in the in situ porcine heart. Experimental Cardiology Group, University of North Carolina at Chapel Hill. AB - INTRODUCTION: The ability to assess transmural changes in action potential duration during acute no-flow ischemia is essential to an understanding of the tachyarrhythmias that occur in this setting. The purpose of this study was to determine if activation recovery intervals determined from unipolar electrograms would provide this information. METHODS AND RESULTS: We recorded simultaneously transmembrane action potentials and unipolar electrograms from sites located as closely together as possible in the center and at the lateral margin of the ischemic zone during acute no-flow ischemia and correlated the changes in activation recovery intervals obtained from the unipolar electrograms to the changes in action potential duration. We found that the activation recovery intervals provided an accurate measure of the changes in action potential duration during acute no-flow ischemia provided the electrograms had a well defined, single negative component to the QRS complex with a maximum negative dV/dt > 10 V/sec and a single positive component to the T wave having a maximum positive dV/dt > 1.6 V/sec. Electrograms meeting these criteria comprised 90% of the electrograms recorded at the margin of the ischemic zone throughout 60 minutes of no-flow ischemia. In the center of the ischemic zone, 75% of the recorded electrograms met these criteria for the first 20 minutes of no-flow ischemia. Thereafter, the percentage declined and after 40 minutes of no-flow ischemia, none of the electrograms recorded in the center of the ischemic zone met these criteria. CONCLUSION: Activation recovery intervals obtained from unipolar electrograms provide an accurate assessment of changes in action potential duration throughout the ischemic zone during acute no-flow ischemia, provided the characteristics of the electrograms meet specific predetermined criteria. PMID- 9727663 TI - Double-wave reentry in orthodromic atrioventricular reciprocating tachycardia: paradoxical shortening of the tachycardia cycle length with development of ipsilateral bundle branch block. AB - INTRODUCTION: Attempts to terminate reentrant tachyarrhythmias by rapid pacing may accelerate the tachycardia. One mechanism for acceleration is double-wave reentry, where two simultaneous wavefronts travel around the same circuit. METHODS AND RESULTS: We report pacing acceleration of AV reciprocating tachycardia (AVRT) due to double-wave reentry in a patient with Wolff-Parkinson White syndrome. The patient had presented with atrial fibrillation and rapid conduction across a left lateral bypass tract. Intravenous procainamide was given during electrophysiologic study because of incessant atrial fibrillation and restored sinus rhythm. Orthodromic AVRT was induced and attempts to terminate the AVRT with right ventricular pacing initiated two alternate tachycardias, both with a left bundle branch block (LBBB) morphology. The first tachycardia, as expected for bundle branch block ipsilateral to the bypass tract, had a longer cycle length (CL) than the original tachycardia (366 msec compared to 297 msec). The second tachycardia had a paradoxically shorter CL, 238 msec compared to 297 msec. Electrogram analysis revealed that the circuit traversed by the accelerated LBBB tachycardia was the same as the slower LBBB tachycardia. The activation sequence revealed two independent wavefronts, traversing this common circuit. As described previously in experimental models, double-wave reentry was initiated when an antidromic-stimulated impulse blocked before colliding with the previous orthodromic impulse, thus allowing two orthodromic impulses to circulate within the circuit. CONCLUSION: We speculate that conduction slowing by procainamide combined with the intrinsic AV nodal delay resulted in the necessary increase in the excitable gap required to develop double-wave reentry. This is the first description of sustained double-wave reentry in humans. PMID- 9727665 TI - Electrical proarrhythmia: induction of inappropriate atrial therapies due to far field R wave oversensing in a new dual chamber defibrillator. AB - This case report describes delivery of atrial therapies during a sinus tachycardia in a new dual chamber implantable cardioverter defibrillator inappropriately caused by far-field oversensing of ventricular beats in the atrial channel. Upon classification of the PR interval pattern, the rate criterion for an atrial tachycardia was fulfilled, and the device initiated high frequency burst pacing as the first stage of programmed tiered atrial therapies. Atrial fibrillation subsequently was induced by high-frequency burst pacing, and eventually a programmed 10-J shock was delivered for successful termination of atrial fibrillation. The phenomenon of far-field oversensing of ventricular beats could be repeatedly observed during exercise testing and abolished by decreasing the atrial sensitivity. PMID- 9727664 TI - Class IC antiarrhythmic drugs, flecainide and pilsicainide, produce ST segment elevation simulating inferior myocardial ischemia. AB - Flecainide and pilsicainide, Class IC antiarrhythmic drugs with slow kinetics, were administered to a 64-year-old man experiencing ventricular tachycardia. Both drugs suppressed the arrhythmia, but caused ST segment elevation in leads II, III, and aVF. No evidence of ischemic heart disease was detected. Withdrawal of the drugs eliminated the ST change. Because these drugs frequently are used to treat tachyarrhythmias in patients who may present with chest pain, this rare ECG manifestation of Class IC drugs should be recognized to avoid misdiagnosis of acute inferior myocardial infarction. PMID- 9727666 TI - A decade of clinical trial developments in postmyocardial infarction, congestive heart failure, and sustained ventricular tachyarrhythmia patients: from CAST to AVID and beyond. Cardiac Arrhythmic Suppression Trial. Antiarrhythmic Versus Implantable Defibrillators. AB - Multiple trials using antiarrhythmic drugs, pharmacologic therapy, and implantable cardioverter defibrillators have been performed in an attempt to improve survival in patients: (1) postmyocardial infarction; (2) with congestive heart failure, with and without nonsustained ventricular tachycardia; and (3) with sustained ventricular tachycardia and those who have survived an out-of hospital cardiac arrest. This article reviews some of the key findings and limitations of completed and ongoing trials. We also make recommendations for the current treatment of such patients based on the results of these trials. PMID- 9727667 TI - Preexcitation in a child with syncope: where is the connection? PMID- 9727668 TI - Clinical strategies in the treatment of atrial fibrillation. Introduction. PMID- 9727669 TI - Mechanisms of atrial fibrillation: mother rotors or multiple daughter wavelets, or both? AB - The mechanism of atrial fibrillation (AF) remains poorly understood. In this article, we present a new unifying hypothesis for the electrophysiologic basis of AF. We surmise that sustained AF depends on the uninterrupted periodic activity of discrete reentrant sites. The shorter reentrant circuits act as dominant frequency sources that maintain the overall activity. The rapidly succeeding wavefronts emanating from these sources propagate through both atria and interact with anatomic and/or functional obstacles, leading to the phenomenon of "vortex shedding" and to wavelet formation. As suggested by recent numerical and experimental results from our laboratory, some of such wavelets may shrink and undergo decremental conduction, others may be annihilated by collision with another wavelet or a boundary, and still others may curl to create new vortices. The end result would be the fragmentation of the periodic wavefronts into multiple independent daughter wavelets, giving rise to new wavelets, and so on in the ceaseless, globally aperiodic motion that characterizes fibrillatory conduction. PMID- 9727670 TI - Mechanisms of pacing interventions in atrial fibrillation. AB - Atrial flutter is due to large single reentrant loops, typically oriented around the caval veins and a line of functional conduction block in between. These reentrant circuits can easily be terminated by properly timed extra beats colliding in the antidromic direction with the reentrant orthodromic wavefront and being blocked in the orthodromic direction within the area of slow conduction. Due to the complex situation in atrial fibrillation with multiple, ever-changing wavelets and a marked functional inhomogeneity of the atrial tissue, these simple concepts do not apply. However, regional control of atrial tissue by rapid pacing is feasible during atrial fibrillation, and through a multisite approach, this pacing modality might lead to a situation where the remaining nonentrained atrial tissue is just no longer reaching the critical mass. Single-site, dual-site, and biatrial pacing seems to reduce the incidence of atrial fibrillation recurrences. Specific advantages and disadvantages of each approach still need to be elucidated. Apart from preventing bradycardia, preventive pacing modes reduce intra-atrial conduction times and might reduce intra-atrial dispersion of refractoriness. Still, the specific mechanisms relevant for their effectiveness need further clarification. PMID- 9727671 TI - Pathogenesis of atrial flutter. AB - It is now known that most cases of atrial flutter are due to reentrant excitation in the right atrium. In the usual reentrant circuit, the reentrant excitation wavefront travels up the interatrial septum and down the right atrial free wall. The boundaries of this reentrant circuit include on one side the tricuspid valve ring and on the other side an area of block, which is probably functional, in the region between the venae cavae. The latter area of block forms during the transitional atrial fibrillation rhythm of variable duration that almost always precedes the initiation of atrial flutter. An isthmus of conduction is also present in the reentrant circuit, and is bounded by the tricuspid ring and the inferior vena cava, the Eustachian ridge, and the coronary sinus. It is probable that an abnormal atrial tissue substrate is usually required. Reentrant circuits around a surgical incision in the atria or around the pulmonary veins (in whole or in part) may be also responsible for atrial flutter. PMID- 9727672 TI - Heart rate variability preceding onset of atrial fibrillation. AB - The purpose of this article is to discuss the relationship between sympathovagal dysfunction and the occurrence of paroxysmal atrial fibrillation. Onset of atrial fibrillation is subject to circadian variation; shorter episodes occur more frequently during the day, whereas longer episodes occur less frequently at night. Vagally mediated atrial fibrillation typically is preceded by bradycardia, is not triggered by stress, occurs more often at night, is more common in men, and occurs at a younger age. Sympathetically mediated atrial fibrillation is less frequent, typically occurs during the day, can be triggered by stress, and often is accompanied by increasing sinus rate and frequent supraventricular extrasystoles. Determination of heart rate variability can be used to evaluate the effects of the autonomic nervous system on sinus rate. The onset of atrial fibrillation at night may be preceded by an increase in high-frequency components of heart rate variability. This is not the case for episodes that occur during the day. The heart rate in sympathetically mediated atrial fibrillation is higher before and during the episode in comparison with the vagal type. Heart rate variability is a promising method for evaluation of the interplay of sympathetic and vagal activity before the onset of atrial fibrillation. However, the role of heart rate variability for diagnostic assessment and therapeutic decision-making in patients with paroxysmal atrial fibrillation remains to be clarified by controlled studies. PMID- 9727673 TI - Signal-averaged P wave: predictor of atrial fibrillation. AB - Abnormalities of the P wave seen during sinus rhythm are associated with atrial fibrillation and other supraventricular arrhythmias. Intra-atrial conduction delays can be seen on the surface ECG as P wave prolongation, which is more visible with averaging techniques used in advanced recording devices. Averaging followed by amplification after proper filtering of the electrical signal should allow more precise measurements of duration and amplitude of the P wave. Data on reproducibility, filter settings, required number of beats, and precise definitions of onset and offset are dependent largely on the system used, which can be QRS or, preferably, P wave triggered. This explains conflicting data in the literature. It is clear that P wave duration is one of the best predictors of perioperative atrial fibrillation. For paroxysmal forms, the data are less convincing. Root mean square voltages of the P wave can be useful. Combining the P wave duration with other data often improves the diagnostic value of signal averaging. Standardization of the techniques appears to be necessary. PMID- 9727674 TI - Atrial dispersion of refractoriness. AB - In contrast to the ventricular myocardium, until recently the electrophysiologic properties of the human atrium had been studied less intensively. Since new, highly effective therapeutic strategies have become available, both clinical and experimental research has been focused on atrial arrhythmias. Experimental data suggest that shortening of the refractory periods and increased dispersion of refractoriness between adjacent areas of the atrium are characteristic for paroxysmal atrial fibrillation. The duration and frequency of atrial fibrillation episodes seem to be associated with persistent electrophysiologic alteration of the atrium. The influence of the autonomic nervous system, antiarrhythmic drugs, and circadian differences also could play a role in the genesis of atrial arrhythmias. PMID- 9727675 TI - Use of intracardiac echocardiography during electrophysiologic evaluation and therapy of atrial arrhythmias. AB - The relationship between endocardial anatomy and the substrate for a variety of atrial arrhythmia mechanisms is being increasingly appreciated. By using intravascular ultrasound imaging systems in the cardiac chambers, direct endocardial visualization can be provided. The advantages include: precise anatomic localization of the ablation catheter tip in relation to important endocardial structures that cannot be visualized with fluoroscopy; the ability to guide ablative procedures partly, or in some instances entirely, by anatomic landmarks; potential reduction in fluoroscopy time; evaluation of stability of catheter tip-tissue contact; confirmation of lesion formation and identification of lesion size and continuity; immediate identification of complications such as clot and pericardial effusion; assistance in the guidance of transseptal puncture; and as a research tool to help in understanding the critical role played by specific endocardial structures in atrial arrhythmogenesis. Presently, intracardiac echocardiography (ICE) is useful as an adjunct in guiding mapping and ablation of focal atrial tachycardia. In our laboratory, it has significant advantage in modification or ablation of sinus node function in patients with inappropriate sinus tachycardia syndrome. Its use in helping to guide ablation of atrial fibrillation remains an exciting, but largely unproved, hypothesis. Better technology will be required if widespread clinical use of ICE is to occur. PMID- 9727676 TI - Multielectrode basket catheter mapping for human atrial fibrillation. AB - In 9 patients with a history of paroxysmal atrial fibrillation (AF), basket catheters with 64 electrodes were used to determine electrophysiologic aspects of onset and early minutes of AF in the right atrium. Mapping was performed using 32 bipolar recordings after induction of AF by programmed atrial stimulation. In 7 patients, registrations were obtained that allowed the onset of AF to be analyzed after the induction. The most common mode of induction was a conduction block and local fragmentation of electrical activation in the area anterior to the tricuspid valve; this was observed in 5 patients. Local cycle lengths (5 min) and correlation dimension of repolarization Dpw_De (2 min) was calculated in 7 patients. The recording sites of the basket catheter were fluoroscopically projected to the anatomic sites of the right atrium. The highest Dpw_Re dimensions as an expression of pronounced chaotic activity were found in the area anterior to the tricuspid valve. PMID- 9727677 TI - Nonfluoroscopic endocardial catheter mapping of atrial fibrillation. AB - The treatment of drug-refractory atrial fibrillation (AF) remains one of the unsolved problems in cardiology. Surgical interventions have demonstrated that AF can be prevented by multiple incisions within both atria. Recently, this strategy has been translated into a catheter procedure. So far, the ablation approach is not based on individual electrophysiologic data, but constitutes only an anatomic approach. Further insight into the spatial and temporal distribution of the local electrograms during AF is needed. Electroanatomic maps acquired by sequential mapping over 45 seconds at each site during AF in six patients with paroxysmal AF were analyzed off-line. Electrograms were sampled at a mean of 36 +/- 12 sites in the left atrium of each patient. A total of 217 sites were sampled, of which 27.3% (59) represented type A (regular) AF, 9.7% (21) represented type B (totally irregular), and 63.1% (137) represented type C (mixture of type A and B) electrograms. The distribution was analyzed in 20 different segments of the left atrium, and a significantly higher incidence of type A electrograms was found in area 3 (upper lateral pulmonary vein) than at all other sites (P < 0.005). This observation needs further confirmation before any conclusion with regard to catheter ablation can be drawn, particularly because the analysis was based on bipolar recordings from a 4-mm tip electrode. PMID- 9727678 TI - Atrial fibrillation in elite athletes. AB - INTRODUCTION: Atrial fibrillation (AF) is a rare event in people younger than 25 years of age, but is probably more frequent in competitive athletes. We analyzed the presence of AF, paroxysmal or chronic, in a population of young elite athletes, including previous Olympic and World champions, who were studied for arrhythmias that endangered their athletic careers. METHODS AND RESULTS: From 1974 to June 1977, 1,772 athletes identified with arrhythmias (1,464 males and 308 females; mean age 21 years) underwent individualized work-ups. Among these, 146 (122 males and 24 females; mean age 24 years) were young elite athletes. They were studied from 1985 to 1997, with a mean follow-up of 62 months. Of the 146 young elite athletes, 13 (9%) had AF (paroxysmal in 11 and chronic in 2); all were male. The paroxysmal AF occurred during effort (n = 7), after effort (n = 1), or at rest (n = 3) and was reinduced by transesophageal pacing or endocavitary electrophysiologic testing under the same clinical circumstances. AF was the cause of symptoms in 13 (40%) of 22 young elite athletes with long lasting palpitations. Five young elite athletes had a substrate for AF: Wolff Parkinson-White syndrome (WPW) in 3, arrhythmogenic right ventricular dysplasia (ARVD) in 1, healed myocarditis in 1, and was considered idiopathic in 8. All elite athletes are alive with a mean follow-up of 62 months and 7 continue in their sports: 3 after radiofrequency catheter ablation (of WPW in 2 and AF with maze-type nonfluoroscopic approach in 1) and 4 after a period of de-training. CONCLUSIONS: AF, occurring in young elite athletes and affecting only males, is one of the most frequent causes of prolonged palpitations and is reproduced easily by transesophageal atrial pacing or electrophysiologic testing. AF may be a cause of disqualification from sports eligibility, but may disappear if the athletic activity is stopped for an adequate period of time, if trigger mechanisms are corrected (i.e., WPW), or if the substrate is modified. PMID- 9727679 TI - Mechanisms of pharmacologic cardioversion of atrial fibrillation by Class I drugs. AB - INTRODUCTION: We recently developed a goat model of sustained atrial fibrillation (AF) in which repetitive induction of AF by burst pacing shortened the atrial effective refractory period (AERP) (electrophysiologic remodeling) and progressively prolonged the paroxysms of AF to become sustained (24 hours) within 1 to 3 weeks (atrial fibrillation begets atrial fibrillation). The aim of the present study was to study the effect of Class I drugs in this animal model of chronic AF. METHODS AND RESULTS: The effects of hydroquinidine (HQ) on seven chronically fibrillating goats and of flecainide (Fl) on nine goats were studied. Both drugs were infused intravenously until sustained AF was cardioverted or adverse drug effects occurred. HQ and Fl restored sinus rhythm in 86% and 67% of the cases. Adverse drug effects occurred in 14% and 56%, respectively. The average atrial cycle length of AF (AFCL) was prolonged to a different degree. Just before restoration of sinus rhythm, the two drugs had increased AFCL by 72% and 50%. The duration of the QRS complex was prolonged 17% by HQ and 50% by Fl. The RR interval was not affected by HQ and was prolonged slightly by Fl. Directly after restoration of sinus rhythm, the AERP during pacing with an interval of 400 msec was 92 +/- 29 (HQ) and 66 +/- 10 msec (Fl) (control value: 149 +/- 10 msec). Intra-atrial conduction velocity was 83 +/- 7 and 86 +/- 11 cm/sec (control value: 116 +/- 10 cm/sec). Although both drugs were effective in terminating AF, after cardioversion the atrial vulnerability was still very high and a single premature stimulus reinduced AF in 100% of the animals. As a result of the short AERP by the AF-induced remodeling and the depressed intra-atrial conduction by the Class I drugs, directly after cardioversion the atrial wavelength was abnormally short (between 5.7 and 7.5 cm). This explains the still high atrial vulnerability to AF directly after cardioversion by Class I drugs. Surprisingly, the prolongation of AFCL by either Class I drug was not due to lengthening of the functional refractory period but rather to a widening of the excitable gap during AF. CONCLUSION: In a goat model of chronic AF, infusion of Class IA and Class IC drugs restored sinus rhythm in 67% to 86% of the cases. However, due to the short AERP and the depressed intra-atrial conduction directly after cardioversion, the atrial vulnerability was still very high and a premature beat easily reinduced AF. PMID- 9727680 TI - Epidemiology and classification of atrial fibrillation. AB - Atrial fibrillation (AF) is a common clinical problem, particularly in the elderly and in patients with organic heart disease. AF generally is classified into paroxysmal and chronic forms. Chronic AF can be the end result of paroxysmal AF in about 30% of patients. Paroxysmal AF can be defined as attacks of arrhythmia lasting < 7 days separated by prolonged periods of sinus rhythm. Chronic AF is AF established for > 7 days. Therefore, the differentiation of paroxysmal from chronic or established AF is based on the history of recurrent episodes and the duration of the current episode of AF. The first episode of persistent AF or the first discovery of AF often is referred to as recent onset AF. Most epidemiologic studies highly underestimate the incidence of paroxysmal and/or asymptomatic AF. The prevalence of AF varies with the age group and patient population studied. AF is found in 3% to 5% of the population > 60 years of age. AF is associated with organic heart disease in 70% to 80% of patients. Of the patients admitted to our Cardiology Division during 1 year, 15% of hospitalized patients had a documented history of AF. The risk of an individual patient developing AF often is difficult to assess, but increasing age and the presence of valvular heart disease and congestive heart failure increase the risk of AF. Other important predictive and causative factors of AF include hypertension, diabetes in women, left ventricular hypertrophy in both sexes, and coronary artery disease, mainly in older patients and patients with left ventricular dysfunction. Other causes of AF include coronary artery disease, hypertrophic cardiomyopathy and dilated cardiomyopathy, chronic obstructive pulmonary disease, pericarditis, and congenital heart disease such as left atrial myxoma and atrial septal. AF can occur in the absence of detectable organic heart disease, so-called "lone AF," in about 30% of cases. The term "idiopathic AF" implies the absence of any detectable etiology including hyperthyroidism, overt sinus node dysfunction, and overt or concealed preexcitation. Stroke is the most important factor of mortality and morbidity associated with AF. These epidemiologic data are essential for designing appropriate therapeutic treatment of this common arrhythmia. PMID- 9727681 TI - Nomenclature of atrial fibrillation or the Tower of Babel. PMID- 9727682 TI - Role of anticoagulant therapy in atrial fibrillation. AB - Atrial fibrillation belongs to the group of cardiovascular diseases that most frequently predispose to arterial thromboembolic events. Within the last years, the AFASAK, BAATAF, SPAF I, SPINAF, and CAFA trials have consistently demonstrated a significant, approximately 70%, risk reduction for stroke on oral anticoagulation in patients with nonrheumatic atrial fibrillation. This benefit by far outweighed the slight increase in annual major hemorrhage. Recently, additional trials (SPAF II, EAFT, SPAF III, and others) have shed further light on important questions concerning risk factors, secondary prophylaxis, the optimal intensity of anticoagulation, and the role of aspirin and other antiplatelet drugs. The main results of these studies are discussed in this review. The majority of patients with atrial fibrillation are > 65 years of age and have other clinical or echocardiographic risk factors. In these patients, adjusted-dose warfarin with target international normalized ratios (INRs) 2.0 to 3.0 is effective and safe. The risk of stroke rises with INR values < 2.0, whereas INR values > 3.0 result in an increase in intracerebral hemorrhages, especially in the very elderly. In contrast, no anticoagulation seems warranted in younger atrial fibrillation patients < 60 years of age without any clinical or echocardiographic risk factor. An overview of all randomized trials that compared aspirin with placebo and/or adjusted-dose warfarin indicates that adjusted-dose warfarin is approximately 50% more effective than aspirin for primary and secondary prevention of stroke, at least in patients with atrial fibrillation who have clinical risk factors. Therefore, oral anticoagulation clearly is the therapy of choice for prevention of thromboembolism in patients with atrial fibrillation. PMID- 9727683 TI - Left atrial appendage function after internal atrial defibrillation. AB - To determine the value of echocardiographic parameters for predicting maintenance of sinus rhythm after internal atrial defibrillation (IAD), transthoracic and transesophageal echocardiography were performed in 38 patients with atrial fibrillation (AF) before IAD. In addition, serial echocardiographic examinations were performed at 1, 7, and 28 days after IAD in 20 patients to assess the effect of IAD on echocardiographic markers of thromboembolic risk. AF had recurred in 49% of patients within 6 months following IAD. Left atrial chamber and appendage size and ejection fraction were not predictive of recurrence of atrial fibrillation. However, peak emptying velocities of the left atrial appendage were significantly lower in patients with recurrence of atrial fibrillation as compared with patients who had maintained sinus rhythm (0.26 +/- 0.1 m/sec vs 0.49 +/- 0.17 m/sec; P = 0.001). A peak emptying velocity < 0.36 m/sec had a sensitivity of 82% and specificity of 83% for predicting the recurrence of AF. Peak A wave velocities increased gradually after cardioversion from 0.47 +/- 0.16 m/sec at 24 hours to 0.61 +/- 0.13 m/sec after 7 days (P < 0.05). One patient developed a new thrombus in the left atrial appendage and another patient suffered a thromboembolic event after IAD. Assessment of left atrial appendage function adds information regarding the probability of maintaining sinus rhythm after the procedure and the need for anticoagulation therapy with IAD. PMID- 9727684 TI - Effects of Class I drugs on atrial fibrillation. AB - This article reviews our knowledge about the efficacy of Class I antiarrhythmic agents, especially quinidine, propafenone, and flecainide, for pharmacologic conversion of atrial fibrillation to sinus rhythm. When given intravenously or orally for the long term, conversion rates between 50% and 90% are reported for restoration of sinus rhythm as well as for maintenance of sinus rhythm after DC cardioversion. Based on transtelephonic monitoring of arrhythmia recurrences as well as tolerance, Class IC agents appear to be especially effective for suppressing clinical symptoms in patients with paroxysmal atrial fibrillation. For patients who develop atrial fibrillation following coronary artery surgery, Class I agents are the second choice of treatment only. The concept of single oral loading with Class IC agents for conversion of atrial fibrillation appears attractive, but more data are needed before we conclude that it is efficacious as well as safe when given to ambulatory patients. Because all Class I antiarrhythmic agents have the potential for lethal proarrhythmia, the greatest and as yet unsettled issue is safety. Until the advent of large-scale and long term trials demonstrating the efficacy and safety of Class I agents for the treatment of patients with atrial fibrillation, this strategy, although very popular to suppress frequent and unpleasant symptoms due to atrial fibrillation, cannot be regarded as firmly established. PMID- 9727685 TI - Effects of Class III drugs on atrial fibrillation. AB - The Class III antiarrhythmic drugs have been used for the treatment of atrial fibrillation (AF); however, each has specific electrophysiologic properties that delineate different safety and/or effectiveness profiles. First-generation Class III agents seem to be more effective in preventing recurrence of AF than in converting AF to sinus rhythm. The high incidence of major cardiac and noncardiac side effects in the long term often requires discontinuation of the chronic antiarrhythmic therapy. The second-generation Class III drugs, ibutilide and dofetilide, have demonstrated interesting clinical applications, especially in the setting of atrial flutter. However, their favorable antiarrhythmic effect is counterbalanced by the high incidence of severe proarrhythmias. New promising experimental data suggest that the new Ikr-ks blockers may be free from these dangerous limitations, thus extending the indication of Class III drugs in the treatment of AF. PMID- 9727686 TI - Atrial fibrillation--maintaining sinus rhythm versus ventricular rate control: the PIAF trial. Pharmacological Intervention in Atrial Fibrillation. AB - INTRODUCTION: In patients with persistent atrial fibrillation, therapy can be directed toward restoration of sinus rhythm or control of ventricular rate only. There are no prospective data in the literature comparing both treatment strategies. METHODS AND RESULTS: Patients with symptomatic persistent atrial fibrillation (duration > or = 7 and < or = 360 days) will be randomized to control of ventricular rate only or to conversion therapy. All patients will be followed for 12 months. Diltiazem is used as the primary therapy for rate control, whereas amiodarone will be administered for restoration and maintenance of sinus rhythm. The primary endpoint of the trial is the recurrence of symptomatic tachyarrhythmic episodes in patients randomized to rate control and recurrent atrial fibrillation in the alternative treatment arm. A number of secondary endpoints also will be investigated. The trial is expected to end its follow-up in the summer of 1998. CONCLUSION: PIAF (Pharmacological Intervention in Atrial Fibrillation) is one of several trials designed to evaluate the optimal treatment strategy in patients with persistent atrial fibrillation. The results hopefully will help to improve care of patients with this common rhythm disorder. PMID- 9727687 TI - Importance of preimplantation procedures in candidates for an implantable atrial defibrillator. AB - INTRODUCTION: Due to the limited efficacy of antiarrhythmic drugs for the treatment of atrial fibrillation, several nonpharmacologic therapeutic options have been developed. One of these options is an implantable atrial defibrillator for patients with severe symptoms and infrequent drug-refractory episodes of atrial fibrillation. The purposes of this study were: (1) to evaluate how many patients with atrial fibrillation are possible candidates for an implantable atrial defibrillator; and (2) to report the results and findings of preimplantation testing in a single center. METHODS AND RESULTS: From our atrial fibrillation outpatient clinic, we evaluated the number of possible candidates for an atrial defibrillator using the following criteria: (1) recurrent persistent atrial fibrillation; (2) long-lasting but infrequent episodes; (3) refractory to antiarrhythmic drugs; (4) capability of maintaining normal sinus rhythm; and (5) no factors increasing proarrhythmic risk. In those patients eligible for an atrial defibrillator, a separate preimplantation test was performed to evaluate atrial defibrillation limits and patient acceptance. Thirty one of 196 patients were possible candidates for an atrial defibrillator. Fourteen of these 31 patients agreed to participate in the METRIX clinical study phase I on atrial defibrillators. Six of these patients met implantation criteria; two patients refused permanent implantation because of intolerable pain. Implantation was performed in four patients; however, one patient could not be cardioverted intraoperatively despite a successful preimplantation test. CONCLUSION: About 16% of selected patients with atrial fibrillation are possible candidates for an atrial defibrillator. However, successful preimplantation testing does not exclude implantation failure. PMID- 9727688 TI - Role of the atrioventricular node in atrial fibrillation: what have we learned from radiofrequency catheter modification of the atrioventricular junction in patients with refractory atrial fibrillation? AB - Determinants of ventricular rate during atrial fibrillation (AF) are controversial. Concealed conduction, summation, rate and irregularity of atrial impulses, pacemaker activity of the AV node, and the electrophysiologic parameters of AV nodal function may all contribute. Findings from series of patients who underwent AV nodal modulation suggest that AV nodal conduction plays some role in the ventricular response to AF. The data are consistent with the presence of dual (or more) atrionodal inputs that play a role in determining AV nodal conduction properties. The data also strongly support the role of summation rather than simple prolongation of refractory periods as a mechanism of ventricular rate control. PMID- 9727689 TI - Radiofrequency ablation of atrial flutter. AB - Atrial flutter can be understood as atrial tachycardia due to a single intra atrial macroreentrant circuit that is determined by fixed or functional boundaries. In various types of atrial flutter, radiofrequency ablation has become an established curative therapy. During the course of an ablation procedure, five steps can be distinguished: (1) determination of the reentrant circuit; (2) identification of the boundaries; (3) proof of the participation of an isthmus between the boundaries in the reentrant circuit; (4) connection of the barriers by a linear lesion; and (5) proof that the line of block is complete. After establishing these five steps, the acute and long-term results of atrial flutter ablation are comparable to those of other supraventricular tachycardias. In this review, we discuss these principles of atrial flutter ablation with an emphasis on typical atrial flutter. PMID- 9727690 TI - Atrioventricular nodal modification and atrioventricular junctional ablation for control of ventricular rate in atrial fibrillation. AB - Atrial fibrillation results in several structural and functional changes in the heart that lead to worsening ventricular function. Although restoration of sinus rhythm is the ideal goal, it is not always feasible. Pharmacologic therapy is associated with adverse effects and is not always effective. We have reviewed the current status of nonpharmacologic therapy in the management of rapid ventricular response due to atrial fibrillation. Electrophysiologic studies have confirmed that the posterior inputs to the AV node have a shorter refractory period and are mainly responsible for maintaining rapid ventricular response in atrial fibrillation. AV nodal modification involves ablation of these posterior inputs in a sequential fashion until a significant reduction of ventricular response is achieved. This procedure has been reported to be successful in maintaining the controlled ventricular response in about 70% of the patients over long-term follow-up. Ablation of the AV node with implantation of a permanent pacemaker is a more definitive procedure and simpler to perform. Reduction in ventricular response achieved with this procedure results in improvement of the patient's clinical symptoms as well as the underlying left ventricular function. Nonpharmacologic therapy for control of ventricular rate should be considered for patients with atrial fibrillation, in whom pharmacologic therapy for rate control is ineffective or poorly tolerated. PMID- 9727691 TI - Surgical treatment of atrial fibrillation. AB - Surgical therapy has been applied in the treatment of atrial fibrillation for almost two decades. At present, the most commonly used approach is the maze operation developed by Cox. In this operation, atrial fibrillation is prevented by critically located incisional lines. Currently, these lines also are drawn during operation using cryoablation or radiofrequency current. To document the value of the maze operation, randomized studies, not only on arrhythmia prevention but also on atrial transport function and thromboembolic complications, should be performed. PMID- 9727693 TI - Intra-atrial defibrillation for atrial fibrillation: animal data. AB - Animal research in the area of atrial defibrillation has helped to bring about a greater understanding of the mechanisms and influencing factors of atrial defibrillation. Increased clinical interest in atrial tachyarrhythmias during the 1980s led to more intensive animal research in this area, in which concepts of intra-atrial cardioversion was only first described in 1974. Initially, the concept of one intracardiac catheter in conjunction with a subcutaneous electrode was evaluated. Further studies involved alternative waveforms, shock durations, electrode materials, and lead configurations to improve the percentage of successful cardioversions while reducing energy requirements. The results from ventricular defibrillation, in which the need for a homogenous field through the majority of fibrillating mass had been established, were applied to research for atrial defibrillation as well. An intracardiac vector with electrodes inside the heart was determined to be most efficient. Results concerning the feasibility, efficacy, and safety have been confirmed by clinical data. Further animal studies will help to evaluate new concepts for reducing energy requirements, such as new electrode materials and advanced shock timing. The clinical benefits of basic research in internal cardioversion of atrial fibrillation justifies additional animal research studies. PMID- 9727692 TI - Multisite electrode pacing for prevention of atrial fibrillation. AB - There is increasing evidence that single site atrial pacing is beneficial for atrial fibrillation (AF) prevention in sick sinus syndrome. Multisite atrial pacing methods such as dual site right atrial pacing and biatrial synchronous pacing are currently under active evaluation for AF and atrial flutter prevention in patients with or without bradyarrhythmias. Clinical studies have demonstrated that multisite atrial pacing has an incremental benefit as compared with single site right and left atrial pacing. The electrophysiologic rationale for the efficacy of multisite atrial pacing is based on the reduction of global and local atrial activation times during pacing and for closely coupled atrial premature beats. This results in earlier recovery of excitability and decreased conduction delay. Dual site right atrial pacing consisting of simultaneous pacing from the high right atrium and the coronary sinus ostium reduces the activation times in virtually all left and right atrial regions, especially in areas of conduction delay. Multisite pacing methods reduce the ability to initiate AF with atrial premature beats by reducing the window for AF induction and minimizing the dispersion of atrial refractoriness. In our long-term clinical experience including 30 patients with paroxysmal and chronic drug-refractory AF, 78% of the patients were free of AF recurrence at 1 year, 63% at 2 years, and 56% at 3 years. Rhythm control was achieved in 86% of patients during a follow-up period of 3 years. Concomitantly, we observed a marked reduction in need for anticoagulation, type I antiarrhythmic drugs, and cardioversion therapies. There were no coronary sinus lead-related complications during follow-up. After the initial favorable clinical experiences, two major prospective randomized trials (DAPPAF and SYNBIAPACE) are under way in North America and Europe to evaluate quantitatively the beneficial impact of multisite atrial pacing for AF prevention. PMID- 9727694 TI - Intra-atrial defibrillation of human atrial fibrillation. AB - Low-energy intra-atrial defibrillation is a new therapeutic option for restoring sinus rhythm in patients with atrial fibrillation (AF). The success rate is quite high when right atrium-coronary sinus or right atrium-pulmonary artery electrode configurations are used, although the former is associated with a slightly lower defibrillation threshold. Several issues regarding the safety and tolerability of the procedure remain unresolved. Our experience and that of others indicate that low-energy intra-atrial cardioversion is safe even during exercise, provided the shock is well synchronized with the QRS and the preshock RR interval is > 500 msec. Reported defibrillation thresholds in patients with persistent AF range from 4 to 8 J, and shocks of this type inevitably are associated with some degree of discomfort. Measures aimed at lowering the defibrillation threshold (e.g., use of biphasic waveforms, antiarrhythmic pretreatment, and use of a single effective shock, as opposed to the multiple shocks delivered in research settings) can be expected to increase patient tolerance and extend the indications for low-energy intra-atrial cardioversion. The procedure currently is indicated for patients with persistent AF who are resistant to external defibrillation unable or unwilling to undergo general anesthesia. The procedure could be expanded to patients affected by obesity in whom the efficacy of external cardioversion is lower. PMID- 9727696 TI - Indications for an atrial defibrillator. AB - Atrial fibrillation is a chronic progressive disease, variable in its expression, but capable of producing considerable symptoms and morbidity. Current therapies are limited in their effectiveness and produce side effects that vary from merely annoying to life threatening. Initial clinical trials of an implanted atrial defibrillator in a selected population have been promising. Early results with patients controlling the conversion of out-of-hospital recurrences of atrial fibrillation suggest that the therapy is well tolerated and successful in converting atrial fibrillation to sinus rhythm. No proarrhythmia has been observed, and the attrition to chronic atrial fibrillation has been very low. PMID- 9727695 TI - Quality of life in patients with atrial fibrillation. AB - INTRODUCTION: The efficacy of a treatment is based primarily on objective criteria such as mortality and morbidity. Besides these criteria, the interest in measuring quality of life in relation to health care has increased in recent years. METHODS AND RESULTS: Although the concept of quality of life inherently is subjective and definitions vary, it generally is agreed that quality of life is a multidimensional construct. The impact of atrial fibrillation (AF) on quality of life has not been evaluated widely using validated methods. Therefore, an international prospective study was designed to assess quality of life over time in patients with AF using validated generic measures and specific conducted disease scales. In addition to a standard demographic questionnaire, patients will complete two predictive scales at baseline and four outcome scales at baseline, and 3-, 6-, and 12-month follow-up. An AF severity score based on subjective and physician-recorded assessments will be used to classify the patient's burden of AF as mild, moderate, or severe. CONCLUSION: Rigorous yet practical approaches are needed to allow for a comprehensive understanding of quality of life in patients with AF. The international study design outlined in this review article represents an attempt to systematically address quality of life in patients with AF and may serve as an example of the types of measures that may be useful in assessing quality of life in patients with AF. PMID- 9727697 TI - Specific considerations with the automatic implantable atrial defibrillator. AB - INTRODUCTION: Internal atrial defibrillation has been evaluated as an alternative approach to the external technique for more than two decades. Previous studies in animals and humans have shown that internal atrial defibrillation is feasible with relatively low energies. The promising results achieved with internal atrial defibrillation have facilitated the development of an implantable atrial defibrillator (IAD). METHODS AND RESULTS: For any new therapy, it is imperative to demonstrate safety, efficacy, tolerability with improvement in quality of life, and cost-effectiveness compared with therapeutic options already available. Maintenance of sinus rhythm or prolonged duration in arrhythmia-free intervals should be demonstrated clearly with an IAD. Initial clinical experience with the Metrix system indicates stable atrial defibrillation thresholds, appropriate R wave synchronization markers, no shock-induced ventricular proarrhythmia, and excellent detection of atrial fibrillation (AF) with a specificity of 100%. Ventricular proarrhythmia has not been reported for correctly R wave synchronized low-energy shocks when closely coupled to RR intervals, and long-short cycles are avoided. CONCLUSION: Preliminary experience with the Metrix system suggests that the IAD may offer a therapeutic alternative for a subgroup of patients with drug refractory, symptomatic, long-lasting, and infrequent episodes of AF. Further efforts must be undertaken to reduce the patient discomfort associated with internal atrial defibrillation in an attempt to make this new therapy acceptable to a larger patient population with AF. PMID- 9727698 TI - Future directions of electrotherapy for atrial fibrillation. AB - Suboptimal treatment of atrial fibrillation (AF) by pharmacotherapy alone has given rise to multiple novel electrotherapeutic modalities including pacing, ablation, and atrial defibrillation. These treatment approaches offer physicians the opportunity to evaluate their patients more extensively to better optimize therapy. AV junctional node ablation followed by full-time pacing improves quality of life and reduces the incidence of tachycardia-induced cardiomyopathy. Physicians who are reluctant to ablate the AV node may opt for either an AV nodal modification procedure or therapeutic modalities that attempt to maintain sinus rhythm. Both biatrial and dual site pacing have met with some success in preventing AF in a limited patient population. Studies also have evaluated the extent to which pacing may be used to terminate AF. These initial studies showed that, in general, pacing was ineffective. Radiofrequency catheter ablation is another therapeutic modality receiving considerable interest. The creation of multiple lesions mimicking the maze procedure, isthmus ablation to prevent the onset of atrial flutter that may degrade into AF, and ablation of tachycardia foci are all potential treatment alternatives to consider either as the primary therapy or as a synergistic procedure designed to augment another electrical therapy. Internal atrial defibrillation is being shown to be both safe and efficacious. This therapy appears uniquely able to repeatedly restore sinus rhythm in a segment of the AF population. Any of these therapies, either alone or more likely in some form of combined therapy, is likely to significantly influence and improve the care of patients with AF in the future. PMID- 9727699 TI - New trials in atrial fibrillation. AB - Large-scale clinical trials in atrial fibrillation (AF) now are addressing issues other than that of thromboembolic prophylaxis. The "rate versus rhythm" debated is fundamental--whether to accept the occurrence of AF and attempt control of the ventricular rate or to strive for the restoration and maintenance of sinus rhythm. The AFFIRM, PIAF, and RACE trials will provide information about the efficacy, costs, adverse effects, and benefits of attempting to maintain sinus rhythm. Atrial pacing is associated with less AF than ventricular pacing, and trials (AFT, SYNBIAPACE, and DAPPAF) now are assessing the antiarrhythmic efficacy of atrial pacing, including dual-site pacing and special algorithms compared with no pacing. PMID- 9727700 TI - Aceruloplasminemia. AB - Aceruloplasminemia is an autosomal recessive disorder of iron metabolism characterized by diabetes, retinal degeneration, and neurologic symptoms. Affected patients evidence marked parenchymal iron accumulation in conjunction with an absence of circulating serum ceruloplasmin and molecular genetic analysis reveals inherited mutations in the ceruloplasmin gene. Taken together with earlier studies that characterized ceruloplasmin as a ferroxidase and recent work indicating an essential role for a homologous multicopper oxidase in iron metabolism in Saccharomyces cerevisiae, these findings reveal an essential role for ceruloplasmin in human iron metabolism. The presence of neurologic symptoms in patients with aceruloplasminemia is unique among the characterized disorders of iron metabolism, and recent findings indicate that astrocyte-specific ceruloplasmin gene expression is critical for iron metabolism and neuronal survival in the retina and basal ganglia. The discovery of this disease provides new insights into the pathways of CNS iron metabolism of direct relevance to a variety of nutritional and genetic disorders of childhood. PMID- 9727702 TI - Measurement of cerebral oxygen consumption in the human neonate using near infrared spectroscopy: cerebral oxygen consumption increases with advancing gestational age. AB - Measurements of cerebral oxygen consumption (CVO2) may improve our understanding of cerebral oxygenation, but there are few published data for sick neonates. Although cerebral maturation is associated with an increase in cerebral glucose consumption, the relationship between CVO2 and increasing gestational age has not previously been assessed in humans. The aims of this study were to evaluate a noninvasive method for the estimation of CVO2 in the neonate using near infrared spectroscopy, and to investigate the relationship between gestational age and CVO2. Twenty babies who were undergoing intensive care in the neonatal period were studied. Cerebral hemoglobin flow (CHbF) and cerebral venous oxyhemoglobin saturation (CSVO2) were measured using near infrared spectroscopy. Arterial oxyhemoglobin saturation was measure by pulse oximetry (SpO2). CVO2 was calculated from the equation: CVO2=CHbF x (SpO2 - SvO2 x 4. The median (range) CVO2 was 0.9 (0.52-1.76) mL x 100 g(-1) min(-1). There was an increase in CVO2 with advancing gestational age (n=20, p=0.55, p=0.014). We conclude that CVO2 can be estimated in sick neonates using noninvasive optical methods. The values obtained are similar to those obtained in other studies by more invasive methods, and are in agreement with values which would be expected from the known rate of cerebral glucose consumption in neonates. Mean (SD) CVO2 at 24-26 wk was 0.5 (0.18) mL x 100 g(-1) min(-1) and rose with increasing gestation to term by 0.03 mL x 100 g(-1) min(-1) per wk. PMID- 9727701 TI - Low dose flunarizine protects the fetal brain from ischemic injury in sheep. AB - Flunarizine, a calcium channel blocker, reduced cerebral damage caused by hypoxic ischemic insults in neonatal rats and in fetal sheep near term. However, the high dose regimen used in these studies produced cardiovascular side effects that might have counteracted the neuroprotective properties of flunarizine. Therefore, the neuroprotective effect was tested in a low dose protocol (1 mg/kg estimated body weight). Twelve fetal sheep near term were instrumented chronically. Six fetuses were pretreated with 1 mg of flunarizine per kg of estimated body weight 1 h before ischemia, whereas the remainder (n=6) received solvent. Cerebral ischemia was induced by occluding both carotid arteries for 30 min. To exclude the possibility that the neuroprotective effects of flunarizine were caused by cerebrovascular alterations we measured cerebral blood flow by injecting radiolabeled microspheres before (-1 h), during (3 min and 27 min) and after (40 min, 3 h, and 72 h) cerebral ischemia. At the end of the experiment (72 h) the ewe was given a lethal dose of sodium pentobarbitone and saturated potassium chloride i.v., and the fetal brain was perfused with formalin. Neuronal cell damage was assessed in various brain structures by light microscopy after cresyl violet/fuchsin staining using a scoring system: 1, 0-5% damage; 2, 5-50% damage; 3, 50-95% damage; 4, 95-99% damage; and 5, 100% damage. In 10 other fetal sheep effects of low dose flunarizine on circulatory centralization caused by acute asphyxia could be excluded. In the treated group neuronal cell damage was reduced significantly in many cerebral areas to varying degrees (range for control group, 1.03-2.14 versus range for treated group, 1.00-1.13; p < 0.05 to p < 0.001, respectively). There were only minor differences in blood flow to the various brain structures between groups. We conclude that pretreatment with low dose flunarizine protects the brain of fetal sheep near term from ischemic injury. This neuroprotective effect is not mediated by changes in cerebral blood flow. We further conclude that low dose flunarizine may be clinically useful as a treatment providing fetal neuroprotection, particularly because the fetal cardiovascular side effects are minimal. PMID- 9727703 TI - Inflammatory pathogenesis of cortical polymicrogyria: an autopsy study. AB - Polymicrogyria, a cortical abnormality usually classified among neuron migration disorders, is characterized by different etiologies and pathogeneses. Recently, it has been proposed that polymicrogyria could be acquired as a consequence of a lasting damage to the developing brain. In this study, we test the hypothesis that an infection in the fetal adnexa may give rise to distant brain defects and eventually polymicrogyria. Thirty-two fetuses spontaneously aborted for extensive ascending chorioamnionitis at 15-26 wk of gestation were evaluated. Control subjects were represented by 8 fetuses aborted at 15-24 wk of gestation. A complete autopsy was carried out between 4 and 12 h after fetal expulsion. We found different histologic alterations in the primitive cortical architecture, both isolated and combined (undulation of the cortical ribbon, untimely cortical folding/molecular layer fusion, and neuronal loss). A total of 25 cases presented one or more of the above-described morphologic alterations in the brain (78%). On the contrary, similar alterations were never observed in any of the control brains (p=0.019). Our findings indicate that chorioamnionitis significantly impairs brain cortex morphogenesis. Such neuron damage may be caused by an unspecific, indirect mechanism of injury to the developing cortex involving hypoxia and free radical generation. The reported brain abnormalities may even evolve into polymicrogyria in surviving fetuses. PMID- 9727704 TI - Do fetal electrocardiogram PR-RR changes reflect progressive asphyxia after repeated umbilical cord occlusion in fetal sheep? AB - The aim of this study was to determine whether there is a relationship between changes in PR-RR correlation of the fetal ECG and progressive changes in fetal acid-base status and blood pressure (BP) during repeated umbilical occlusion. Chronically instrumented fetal sheep at 126.8+/-0.6 d (mean+/-SEM) were randomized to receive 1 min of total umbilical cord occlusion either every 5 min for 4 h (1:5 group; n=8), or every 2.5 min until BP fell <2.7 kPa (20 mm Hg) on two successive occlusions (1:2.5 group; n=8). The PR-RR correlation was determined in 5- or 2.5-min intervals. Umbilical cord occlusion caused variable decelerations with initial sustained hypertension. In the 1:5 group BP remained elevated throughout, and there was little change in acid-base status (pH=7.34+/ 0.07, base deficit=1.3+/-3.9 after 4 h). In contrast, after the third occlusion the 1:2.5 group showed progressive hypotension during occlusions, and severe progressive metabolic acidemia (pH 6.92+/-0.1, base deficit 17.0+/-4.7 mmol/L after the last occlusion). In both groups, the PR-RR relationship switched from positive to negative with the onset of occlusions, then reverted to positive after a variable interval. In the 1:2.5 group later reversion of the PR-RR to positive was associated with earlier and more prolonged hypotension during the middle and end of the occlusion series (p < 0.001). We conclude that the initial switch to a negative PR-RR relationship during repetitive umbilical occlusion was due to a reflex-mediated response unrelated to fetal acidosis or hypotension. Both stable well compensated fetuses and severely hypoxic, hypotensive fetuses subsequently showed a positive PR-RR correlation. PMID- 9727705 TI - Persistent increases in cerebral lactate concentration after birth asphyxia. AB - In this prospective study proton magnetic resonance spectroscopy (1H MRS) was used to test the hypothesis that lactate can be detected later than 1 mo after birth in the brains of infants who display severe neurodevelopmental impairment 1 y after transient perinatal hypoxia-ischemia. Data were obtained from three groups of infants: 1) eight infants suffering birth asphyxia followed by perinatal encephalopathy and abnormal neurodevelopmental outcome at 1 y of age (defined as major neurologic impairment, Griffiths quotient <85%, and low optimality score); 2) 10 infants with signs of perinatal hypoxia-ischemia but normal neurodevelopmental outcome at 1 y; and 3) six control infants with uneventful perinatal courses and normal neurodevelopment at 1 y. Between one and four examinations (median 1) were performed at median (range) 11 (4-68) wk after birth, and the cerebral concentration ratio of lactate to creatine plus phosphocreatine (Cr) calculated from each spectrum. Lactate was detected later than the 1st mo after birth in seven of eight infants with abnormal neurodevelopmental outcome [maximum detected lactate/Cr was median (range) 0.44 (0.24-0.67)]. No lactate was detected later than the 1st mo after birth in infants with normal neurodevelopmental outcome, nor in five of six control subjects, although a small amount of lactate was detected in one control infant (lactate/Cr=0.04). These results suggest that the pathologic postasphyxial process, indicated by persistent cerebral lactate, may not be confined to the period immediately after injury. PMID- 9727706 TI - Bilirubin inhibits transport of neurotransmitters in synaptic vesicles. AB - Uptake of neurotransmitters into synaptic vesicles occurs through specific transport proteins which are driven by an ATPase-generated electrochemical force consisting of a proton gradient and a membrane potential. In this study we examined the effects of bilirubin, a well known neurotoxic agent, on the vesicle uptake both of [3H]dopamine (which is driven mostly by the proton gradient) and [3H]glutamate (which is driven mostly by the membrane potential), and compared these to the vesicular proton gradient, which was estimated by analyzing the uptake of [14C]methylamine. Bilirubin inhibited the uptake of both dopamine and glutamate (p < 0.01), with an identical dose-response curve for both transmitters. Inhibition was detected readily at 75 microM. The effects of bilirubin were dependent on the concentration of vesicles in the assay, suggesting that the concentration of bilirubin in the membranes and not the water phase was important. Bilirubin also decreased uptake-dependent efflux of dopamine from the vesicles. In contrast, bilirubin had no effect on the vesicular proton gradient, as measured by methylamine uptake. Our results show that bilirubin has essentially identical inhibitory effects on the uptake of both a monoamine transmitter and an amino acid transmitter into synaptic vesicles, but does not influence the vesicular H+-ATPase or proton translocation. Our data suggest an inhibitory interaction between bilirubin and several transport proteins in synaptic vesicle membranes. PMID- 9727707 TI - Prenatal dexamethasone causes oligonephronia, sodium retention, and higher blood pressure in the offspring. AB - Recent reports have shown that low birth weight infants have a higher incidence of adult hypertension. These observations have stimulated a number of studies designed to evaluate the mechanisms of this phenomenon. In this study, fetal growth retardation was induced by treating pregnant rats with dexamethasone. After birth, pups whose mothers were treated with dexamethasone had a lower body and kidney weight and a lower number of glomeruli than control pups. Immunohistochemistry on treated kidneys demonstrated a marked reduction in the number of cells undergoing mitosis in the cortical nephrogenic zone. In the treated group, body and kidney weight normalized by 60 d of age, but blood pressure was significantly higher compared with controls (130+/-4 versus 107+/-1 mm Hg). In addition, GFR was significantly lower, albuminuria was higher, urinary sodium excretion rate and fractional sodium excretion were lower, and sodium tissue content was higher. In contrast, when pregnant rats were treated with a natural glucocorticoid (hydrocortisone) which is metabolized by the placenta, fetal development and adult blood pressure were normal. In conclusion, we found that high levels of maternal glucocorticoids impair renal development and lead to arterial hypertension in offspring. Even though renal mass eventually normalizes, glomerular damage as well as sodium retention occur and these factors may contribute to the development of hypertension. PMID- 9727708 TI - Ontogeny and regulation of cardiac angiotensin types 1 and 2 receptors during fetal life in sheep. AB - Previous studies have shown that the expression of cardiac angiotensin II (ANG II) type 1 (AT1) and type 2 (AT2) receptors are developmentally regulated, although factors modulating these receptors have not been well investigated. The present study was designed 1) to characterize the ontogeny of cardiac AT1 and AT2 gene expression during the last third trimester of gestation in fetal sheep and newborn lambs, 2) to determine the influence of ANG II on modulating cardiac AT1 and AT2 gene expression during fetal life, and 3) to investigate the role of AT1 receptor activity on the regulation of AT1 and AT2 mRNA levels during fetal cardiac development. Using sheep AT1 and AT2 cDNA probes, we demonstrated that cardiac AT1 gene expression is relatively unchanged during fetal (90-135 d of gestation, term 145 d) and newborn life. In contrast, cardiac AT2 mRNA expression was high during fetal development and decreased rapidly after birth. Continuous i.v. infusion of ANG II (9.5 nM/h) for 24 h, which raised ANG II levels from 84+/ 9 to 210+/-21 pg/mL had no effect on the expression of cardiac AT1 or AT2 mRNA, but increased adrenal and decreased liver AT1 mRNA levels. Administration of the AT1 receptor antagonist losartan (1.2 mg kg(-1) h(-1)) significantly decreased arterial blood pressure in fetuses at 110- and 135-d, but not 95-d gestation. Except for increased AT1 receptor gene expression in the right atrium at 95- and 135-d gestation, and left ventricle at 110-d gestation, cardiac AT1 and AT2 mRNA levels were unaltered by AT1 receptor blockade. In summary, this study demonstrates that cardiac AT2 but not AT1 receptor gene expression is regulated by the transition from fetal to newborn life. Neither ANG II nor blockade of AT1 receptors significantly alter the expression of AT1 or AT2 mRNA in the fetal heart. Endogenous ANG II also appears to significantly contribute to the maintenance of blood pressure homeostasis during the final third of gestation in fetal lambs. PMID- 9727709 TI - Comparison of pulmonary inflammatory mediators in preterm infants treated with intermittent positive pressure ventilation or high frequency oscillatory ventilation. AB - Ventilated preterm infants prone to the development of bronchopulmonary dysplasia have been shown to have increased inflammatory mediators in their tracheal aspirates. High frequency oscillatory ventilation (HFOV) is thought to be less traumatic than intermittent positive pressure ventilation (IPPV) in premature infants with surfactant deficiency, and therefore may reduce the inflammatory response in tracheobronchial aspirates. We randomized 76 premature infants requiring mechanical ventilation (birth weight 420-1830 g, median 840 g, gestational age 23 3/7 to 29 2/7 wk, median 26 4/7 to receive either an IPPV with a high rate (60-80/min) and low peak pressures, or an HFOV aiming at an optimization of lung volume, within 1 h of intubation. Tracheal aspirates were systematically collected during the first 10 d of life and analyzed for albumin, IL-8, leukotriene B4 (LTB4), and the secretory component (SC) for IgA as a reference protein. Bacterially colonized samples were excluded. On the treatment d 1, 3, 5, 7, and 10, the resulting median values of albumin (milligrams/mg of SC) were 28, 23, 24, 18, and 10, in IPPV-ventilated infants, and 33, 28, 18, 25, and 39 in HFOV-ventilated infants, respectively. Median IL-8 values (nanograms/mg of SC) were 671, 736, 705, 1362, and 1879 (IPPV) and 874, 1713, 1029, 1426, and 1823 (HFOV), respectively, and median LTB4 values (nanograms/mg of SC) were 26, 13, 27, 22, and 11 (IPPV) and 15, 12, 7, 12, and 16 (HFOV), respectively. Values were similar in IPPV- and HFOV-ventilated infants, and no significant differences were noted. We conclude that HFOV, when compared with a high rate low pressure IPPV, does not reduce concentrations of albumin, IL-8, and LTB4 in tracheal aspirates of preterm infants requiring mechanical ventilation. PMID- 9727711 TI - Major vasodilator role for nitric oxide in the gastrointestinal circulation of the mid-gestation fetal lamb. AB - As nitric oxide (NO) may be a particularly important vasodilator in early life, we investigated its role in the regulation of the gastrointestinal (GI) circulation at mid-gestation. Cardiac output and GI blood flow were measured by the radioactive microsphere technique in eight chronically instrumented and unanesthetized mid-gestation fetal sheep. Mean arterial pressure (MAP), heart rate, blood flow, oxygen delivery, and vascular resistance were determined before and after infusion of the specific NO synthase inhibitor, Nomega-nitro-L-arginine (L-NNA) at doses of 10 and 25 mg/kg. In response to L-NNA infusion, MAP increased (p < 0.01) and combined ventricular output decreased (p < 0.001). GI blood flow and oxygen delivery decreased and vascular resistance increased in the stomach and all segments of the small and large intestine (all p < 0.001). The greatest reduction in blood flow was in the small intestine (p < 0.01) and the basal differential pattern of small intestinal blood flow exceeding large intestinal flow was completely abolished. These changes were much greater than those previously described in late-gestation fetuses. Our results suggest that, at mid gestation, NO plays a major role in the regulation of blood flow and vascular tone across all segments of the fetal GI tract, with its effects being more pronounced than later in development. PMID- 9727710 TI - Decreased gene expression of endothelial nitric oxide synthase in newborns with persistent pulmonary hypertension. AB - Previous studies in adults have shown that chronic pulmonary hypertension is associated with decreased endothelial nitric oxide synthase (eNOS) expression in pulmonary arteries. However, the role of decreased eNOS expression in persistent pulmonary hypertension of the newborn (PPHN) is unknown. We investigated the hypothesis that umbilical vein endothelial cells cultured from infants with PPHN will have decreased eNOS expression. Umbilical cords were collected from meconium stained infants at birth, and endothelial cells were isolated if the infants developed PPHN. Endothelial cells were grown in primary culture, and total RNA was isolated. cDNA was reverse transcribed from mRNA and amplified by PCR. An expected product of approximately 550 bp was found in all control infants but only in two of the six infants with PPHN. Identity of the PCR product was confirmed by Southern hybridization to a separate internal eNOS-specific probe. Amplification of beta-actin cDNA, an internal control, was detected in all controls and in all infants with PPHN, including the four infants without the eNOS band. There was no difference in the course and outcome of patients with presence or absence of the eNOS band. However, there was an acidotic arterial blood pH (7.19-7.29) and intrapartum fetal heart rate decelerations in all four infants without eNOS expression. In conclusion, eNOS mRNA was detected in all normal term infants but was notably absent in the majority of infants with PPHN in this pilot study. The development of PPHN is multifactorial, and a decrease in eNOS gene expression may occur in some infants. Whether the decreased eNOS transcript is a cause of PPHN or a result of intrapartum stress remains to be determined. PMID- 9727712 TI - The relationship of antiphospholipid antibodies to thromboembolic events in pediatric patients with systemic lupus erythematosus: a cross-sectional study. AB - The purpose of this study was to evaluate pediatric patients with systemic lupus erythematosus (SLE) to determine 1) the incidence of thrombosis, 2) the incidence of antiphospholipid antibodies, and 3) whether there is an association between the presence of antiphospholipid antibodies and thrombosis. We performed a cross sectional cohort study in 59 consecutive SLE patients who had been managed at rheumatology clinics in two pediatric hospitals. A history, questionnaire, and chart review were completed by the study nurse blinded to laboratory results. Only the thrombotic events that could be substantiated by review of radiographic tests were accepted. The presence of antiphospholipid antibodies was determined by prospective analysis for a lupus anticoagulant and anticardiolipin antibodies on two separate occasions at least 3 mo apart. Patients were considered to be positive if one or more tests were positive on both occasions. Thirteen thrombotic events occurred in 10 of the 59 patients (17%). Fourteen patients (24%) were classified as positive for lupus anticoagulant, and 19 patients (27%) were classified as positive for anticardiolipin antibodies. A significant relationship between the presence of a lupus anticoagulant and a thrombotic event was shown: odds ratio 28.7 (95% confidence interval 4.03-138.2, p < 0.001). A nonsignificant trend was seen for the presence of an anticardiolipin antibody and a thrombotic event: odds ratio 2.12 (95% confidence interval 0.71-22.8, p=0.08). We conclude that in pediatric patients with SLE: 1) a significant proportion of patients have thrombotic events, 2) a significant proportion of patients have antiphospholipid antibodies, and 3) there is a significant relationship between the presence of a lupus anticoagulant and thrombotic events. PMID- 9727713 TI - Elevation of metabolic rate by pyrogen administration does not affect the gain of respiratory peripheral chemoreflexes in unanesthetized kittens. AB - We previously reported that reducing environmental temperature from 30 to 25 degrees C increases the gain of respiratory chemoreflexes. To investigate the role of increased metabolism in mediating the effect on the gain of the respiratory chemoreflex, we compared the respiratory responses, at ca. 26 degrees C to breath-by-breath alternations of inspired gas between air and 14% oxygen (hypoxia run) or air and 5% CO2 (CO2 run) with that to alteration of air between two inspired lines (control run) before and after the injection of a pyrogen (IL 1beta 400 ng/kg i.p.) in eight kittens at 27-35 d of postnatal age. The respiratory chemoreflex was quantified from the alternations in inspiratory and expiratory variables produced during test runs in terms of the direction and the amplitude of the alternation for each variable and compared with the results of control runs at the same temperature. Pyrogen administration produced a rise in rectal temperature and in oxygen consumption. However, there was no difference in the chemoreflex response to hypoxia or CO2 runs, in terms of either the pattern or of the amplitude of alternation, before and after the injection of the pyrogen. We conclude that the increase in the gain of chemoreflex observed during cooling in a previous study is not due to an increase in metabolism. Some change in input from thermoreceptors may bias the gain of chemoreflexes. PMID- 9727714 TI - Exhaled isoprene and acetone in newborn infants and in children with diabetes mellitus. AB - A new analytical method gas chromatography combined with UV spectrophotometry was used to measure isoprene and acetone in expired breath collected from four different groups of children: 1) healthy newborn babies, 2) healthy preschool children, 3) healthy school children, and 4) diabetic children in different metabolic states. Both isoprene and acetone could readily be determined in one single analysis of a 250-mL air sample. Newborn babies during the first postnatal week had undetectable or very low levels of isoprene in their expired air irrespective of catabolic or anabolic state. Breath isoprene increased with age, and healthy school children had higher levels than did healthy preschool children. No significant differences in breath isoprene were found between healthy and diabetic children. Breath acetone was found to correlate with metabolic state both in newborn babies and in diabetic children. These findings illustrate the potential use of a new technique for breath analysis in children with metabolic disturbances. PMID- 9727715 TI - Maternal, umbilical, and amniotic fluid concentrations of tryptophan and kynurenine after labor or cesarean section. AB - A major route of tryptophan metabolism is via the hepatic and cerebral synthesis of kynurenine, a substance subsequently used by astrocytes in the brain for the production of the neuroactive substances kynurenic acid and quinolinic acid. Both kynurenic and quinolinic acids have been implicated in modulating the activity of excitatory amino acid pathways in the brain, the former as a neuroprotectant because of its antagonist properties, and the latter as an excitotoxin because of its agonist actions, at NMDA receptors. We therefore determined the concentrations of tryptophan and kynurenine in maternal venous and umbilical cord blood, and in amniotic fluid, of infants after labor and vaginal delivery, and after delivery by cesarean section. Concentrations of tryptophan and kynurenine were significantly higher in umbilical vein plasma compared with maternal venous plasma. Tryptophan and kynurenine concentrations in umbilical vein plasma and amniotic fluid were significantly higher after labor, compared with samples obtained from infants of the same gestational age delivered by cesarean section. There was no umbilical vein-to-artery concentration difference for kynurenine in samples obtained after either labor or cesarean section, but there was a significant gradient for tryptophan in samples obtained after vaginal delivery, indicating increased transfer of this amino acid during labor. There was a significant correlation between umbilical vein tryptophan and kynurenine concentrations for both the labor and cesarean section groups, and plasma kynurenine concentrations were also significantly correlated with both umbilical vein cortisol concentrations and the duration of the second stage of labor in the vaginally delivered infants. These results suggest that the placental transfer of tryptophan and the fetal synthesis of kynurenine are increased during labor. These findings have implications for understanding the vulnerability of the infant brain to ischemic/hypoxic damage in the perinatal period. By analogy with the adult brain, the molar ratio of these substances is likely to determine the susceptibility of the brain to seizure and excitotoxic damage. PMID- 9727716 TI - Intrauterine growth retardation: evidence for the activation of the insulin-like growth factor (IGF)-related growth-promoting machinery and the presence of a cation-independent IGF binding protein-3 proteolytic activity by two months of life. AB - Thirty-seven children with intrauterine growth retardation (IUGR) were enrolled in a 3-mo longitudinal study. Weight, length, and knee-heel length (by knemometry) were measured at birth and at 7, 14, 30, 60, and 90 d. GH, IGF-I, IGF binding protein (BP)-3, IGFBP-1, and C-peptide were measured at birth and at 2 mo. IGFBP-3 Western immunoblotting and proteolytic activity assay were also performed. Twenty-five newborns with birth weight appropriate for gestational age were chosen as controls. At birth IUGR newborns showed levels of GH and IGFBP-1 significantly higher, and IGF-I, IGFBP-3, and C-peptide significantly lower than control subjects. At 2 mo GH and IGFBP-1 levels decreased, whereas IGF-I, IGFBP 3, and C-peptide rose, attaining the concentrations found in control subjects at birth. Baseline peptide levels as well as their 2-mo variations did not correlate with the gain in weight, supine length, and knee-heel length recorded at 3 mo. Fourteen of nineteen IUGR cord blood samples showed the presence of the intact approximately 42-39-kD IGFBP-3 doublet and the major approximately 29-kD fragment. At 2 mo the IGFBP-3 band pattern was characterized by the predominance of a approximately 18-kD fragment in 6 of 19 tested IUGR infants. The incubation of 2-mo IUGR samples with normal adult serum induced the appearance of the approximately 18-kD band, which was not modified by the addition of EDTA. These results suggest that: 1) the IGF-related growth-promoting mechanism is impaired in IUGR children at birth but is fully restored at 2 mo; 2) the cord blood levels of GH, IGF-I, IGFBP-3, IGFBP-1, and C-peptide are not predictive of the weight and length gain during the first 3 mo of life; 3) IUGR children have at least two different IGFBP-3 proteases, one cation-dependent protease that is present at birth and able to yield the major approximately 29-kD IGFBP-3 fragment and a second one, with a different activation timing, which exhibits cation independence and induces the formation of a approximately 18-kD IGFBP-3 form. PMID- 9727717 TI - Gyrate atrophy of the choroid and retina: lymphocyte ornithine-delta aminotransferase activity in different mutations and carriers. AB - Deficiency of omithine-delta-aminotransferase (OAT) causes gyrate atrophy of the choroid and retina with hyperornithinemia (GA; McKusick 258870), a progressive autosomal recessive chorioretinal degeneration leading to early blindness. As residual enzyme activity may vary in different mutations of the OAT gene and explain individual variations in disease progression, a sensitive HPLC modification of the OAT assay in lymphocytes was developed, based on measurement of the dihydroquinozolinium reaction product. The OAT activities (ranges) of 43 Finnish GA patients with mutations L402P/L402P, R180T/L402P, N89K/ L402P, and L402P/x (x = previously unknown allele), were <1-10, <1-13, <1-17, and <1 pmol x min(-1) mg protein(-1), respectively. The OAT activities (mean+/-SD) of nine L402P/ wild heterozygotes were 70+/-50 (range 33-193), and those of 15 healthy control subjects 184+/-60 (range 85-291) pmol x min(-1) mg protein(-1). This lymphocyte assay is an easy, rapid, and sensitive method for reliable recognition of GA homozygotes. OAT mutations of the Finnish patients show similar residual enzyme activity in the lymphocytes. OAT activities in the L402P heterozygotes and healthy control subjects overlap, suggesting that, for reliable carrier detection, the OAT alleles have to be studied. However, as all OAT mutations are not known, direct measurement of enzyme activity has a role in heterozygote identification and possibly also in prenatal diagnosis of GA. PMID- 9727718 TI - Urinary organic acids in infant malnutrition. AB - The metabolic derangements in severe protein-energy malnutrition (PEM) are only partially known, due to the limitations of blood collection in these patients. Urinary excretion of organic acids was studied by gas chromatography-mass spectrometry in 39 infants with four types of PEM: 1) upon hospital admission, as soon as eventual infections had been cleared, and salt and water deficits corrected, but before oral feeding was started; 2) after start of protein alimentation; 3) on the day of discharge. All of the patients showed an increased excretion of various organic acids at some point of their hospital stay, regardless of the clinical type of PEM. In nearly half of the malnourished children, results were suggestive of blocks in the pathways of propionate (15.4% with increased methylmalonate and 25.6% with 2-methylcitrate), of fatty acid beta oxidation (30.8% with raised dicarboxylic acids with low or low normal 3 hydroxybutyrate), or of both pathways (12.8%). These abnormalities may have been caused by cofactor deficiencies (biotin, vitamin B12, riboflavin, carnitine, niacin). Dicarboxylic acids were excreted in high amounts since the initial sample, probably due to increased mobilization of fatty acids. Increased 2 methylcitrate and methylmalonate excretion was observed more frequently once patients started to be orally fed. The accumulation of potentially toxic acyl-CoA precursors of these compounds could contribute to the known clinical worsening of some malnourished infants after suddenly increased protein intake. Other less specific metabolites, such as 3-hydroxybutyrate, lactate, 4-hydroxyphenyllactate, fumarate, succinate, and 4-hydroxyphenylacetate, were also abnormally excreted in some patients. The analysis of urinary organic acids provides a new approach for the metabolic study of PEM and may have diagnostic and therapeutic implications. PMID- 9727719 TI - Hepatic mitochondrial 3-hydroxy-3-methylglutaryl-coenzyme a synthase deficiency. AB - There are at least two isoenzymes of 3-hydroxy-3-methylglutaryl (HMG)-CoA synthase (EC 4.1.3.5) located in the mitochondrial matrix and the cytoplasm of hepatocytes, respectively. The mitochondrial enzyme is necessary for the synthesis of ketone bodies, which are important fuels during fasting. We report a child with a deficiency of this isoenzyme. He presented at 16 mo with hypoglycemia. There was no rise in ketone bodies during fasting or after a long chain fat load but there was a small rise after a leucine load. Measurement of beta-oxidation flux in fibroblasts was normal. Using antibodies specific for mitochondrial HMG-CoA synthase, no immunoreactive material could be detected on Western blotting. Total HMG-CoA synthase activity in liver homogenate was only slightly lower than in control samples. Presumably, as there was no mitochondrial HMG-CoA synthase enzyme protein, this activity arose from the cytoplasmic or other (e.g. peroxisomal) isoenzymes. With avoidance of fasting, our patient has had no problems since presentation and is developing normally at 4 y of age. PMID- 9727720 TI - Effect of oxypurinol, a xanthine oxidase inhibitor, on hepatic injury in the bile duct-ligated rat. AB - Oxidant stress has been implicated as playing a role in the pathogenesis of cholestatic liver injury. The objective of this study was to determine whether the xanthine oxidase/xanthine dehydrogenase enzyme system was involved in this oxidant stress. Adult Sprague-Dawley rats were treated with the xanthine oxidase inhibitor, oxypurinol, and randomized to bile duct ligation or sham surgery; vehicle-treated, sham-operated rats served as controls. After 5 d of bile duct ligation, serum aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, and total and direct bilirubin concentrations were significantly elevated, and increased lipid peroxidation of hepatic mitochondria and microsomes was present. Treatment with oxypurinol reduced the aspartate aminotransferase, alanine aminotransferase, and bilirubin values by 26-47% but did not alter the increased lipid peroxidation of mitochondria and microsomes. Serum vitamin E:total lipids ratio was also reduced in both bile duct-ligated groups, consistent with oxidant injury. These data show that inhibition of xanthine oxidase reduces biochemical evidence of hepatocellular injury during bile duct ligation without affecting oxidant damage to intracellular hepatocyte organelles. Thus, in this model a component of cholestatic injury appears to have been caused by oxidant stress from a source outside of the hepatocyte. PMID- 9727721 TI - Transforming growth factor-beta1 in plasma and liver of children with liver disease. AB - Although several liver diseases of childhood, particularly biliary atresia (BA) and cystic fibrosis (CF) liver disease (CFLD) are characterized by hepatic fibrosis, the pathogenesis of this process is incompletely understood. The cytokine transforming growth factor-beta1 (TGF-beta1) has been implicated in hepatic fibrosis in experimental animals, in which both the hepatic expression and plasma concentration of this cytokine are increased. The objective of our study was to determine whether there are similar alterations of TGF-beta1 in patients with hepatic fibrosis secondary to either BA and/or CFLD. The study design was as follows. In study 1, plasma TGF-beta1 was assessed by ELISA in 9 children with BA undergoing liver transplantation, 11 patients with CFLD, and appropriate control subjects. In study 2, hepatic expression of TGF-beta1 protein (assessed immunohistochemically) and hepatic fibrosis were scored semiquantitatively, on a 1-3 scale, by blinded investigators, in archival liver biopsy specimens from 10 children with BA, 10 with CFLD, and from 10 older children with normal hepatic histology, as well as in 4 patients with liver diseases of various etiologies. Simultaneous plasma and liver TGF-beta1 studies were performed in 8 patients with liver disease. Results were as follows. Plasma TGF-beta1 values were inversely correlated with age in healthy subjects (r=-0.54, p < 0.0001). The plasma TGF-beta1 protein of children with BA was decreased (13+/ 2 ng/mL) compared with values for healthy children (42+/-6 ng/mL, n=10, p < 0.005). Similarly, the plasma TGF-beta1 concentration in patients with CFLD was also decreased compared with values for children with CF and normal serum liver profiles (n=14) (2+/-1 ng/mL versus 12+/-1, p < 0.05). However, the plasma TGF beta1 concentration was increased in two patients with other types of liver disease. The hepatic expression of TGF-beta1 was increased in the presence of hepatic fibrosis in all types of liver diseases studied. Forty-six percent of patients had both marked hepatic fibrosis and marked TGF-beta1 labeling; 86% of samples without fibrosis showed no TGF-beta1 labeling, p=0.007. In conclusion, these studies have established the association of hepatic TGF-beta1 protein and hepatic fibrosis in several common liver diseases of childhood. Our data also suggest that, in children, plasma TGF-beta1 does not appear to be a useful marker of hepatic expression of this cytokine. PMID- 9727722 TI - Creatine increases survival and suppresses seizures in the hypoxic immature rat. AB - The incidence of clinical seizures is highest in the newborn period. At this developmental stage seizures have many causes, with hypoxia and ischemia thought to be the most common. In rat pups hypoxia produces seizures most frequently at 10-12 d of age. Brain cellular energy metabolism increases between 5 and 25 d of age in the rat, as indicated in vivo by the phosphocreatine (PCr)/nucleoside triphosphate (NTP) ratio measured by 31P nuclear magnetic resonance (NMR) spectroscopy. Brain PCr/NTP ratios are approximately the same in 10-12-d-old rats and human term newborns, the ages of high seizure susceptibility. Thus, low Cr or PCr may be important in susceptibility to hypoxic seizures in the metabolically immature brain. To test this hypothesis, rat pups were injected with Cr for 3 d before exposing them to hypoxia on postnatal d 10 or 20. Before and during hypoxia, the electrocortical activity or 31P nuclear magnetic resonance spectra were measured. At 10 but not 20 d, Cr injections increased brain PCr/NTP ratios, decreased hypoxia-induced seizures and deaths, and enhanced brain PCr and ATP recoveries after hypoxia. Thus, Cr protects the metabolically immature brain from hypoxia-induced seizures and, perhaps, from cellular injury. These results may be directly relevant to the human newborn. PMID- 9727723 TI - The teratogenicity of N-methyl-D-aspartate (NMDA) receptor antagonists. PMID- 9727724 TI - The teratogenicity of N-methyl-D-aspartate (NMDA) receptor antagonists. PMID- 9727725 TI - Vitamin E in the treatment of tardive dyskinesia: a preliminary study over 7 months at different doses. AB - Vitamin E has been suggested to be a promising new treatment for tardive dyskinesia. However, little is known about the optimum dose. Twenty patients with tardive dyskinesia whose medication had been unchanged for at least 1 month were selected and randomly divided into a treatment group with 11 patients and a control group with nine patients. The treatment group was started on 600 mg of vitamin E per day, and this dose was increased over the 7 months of the trial to 1600 mg per day. The medication for the control group was unchanged. Severity of tardive dyskinesia was rated on the Abnormal Involuntary Movement Scale. Patients in the treatment group initially showed a significant response to the lower dose of 600 mg per day. However, this improvement was not maintained and differences between the two groups reached significant levels only after the dose of vitamin E was increased to 1600 mg per day. At this dose, there was a significant and sustained reduction in the severity of tardive dyskinesia. The results suggest that vitamin E is of value in the treatment of tardive dyskinesia and that the optimum dose for treating tardive dyskinesia is 1600 mg per day. In addition, there may be a dose related therapeutic effect of Vitamin E in tardive dyskinesia. PMID- 9727726 TI - Safety and tolerance of zolpidem in the treatment of disturbed sleep: a post marketing surveillance of 16944 cases. AB - The subjective response to treatment with zolpidem, an imidazopyridine hypnotic, was assessed in patients with insomnia under normal treatment conditions in an outpatients' practice in Germany. The uncontrolled clinical surveillance study included 16944 outpatients with subjective difficulties in initiating and/or maintaining sleep. Office-based neurologists, psychiatrists, internists and general practitioners were asked individually to adjust the dosage of zolpidem (age < or = 65 years, 10-20 mg; age > 65 years, 5-10 mg) over a recommended period of 3-4 weeks. In total, 82.8% of patients completed the survey (36% men, 64% women, mean age 58.8 +/- 14.9 years; 58.6% without previous hypnotic medication; duration of sleep complaints > 6 months in 40.6%, 1-6 months in 27.8%). Most patients (63.9%) took zolpidem on a daily basis. The average dose was 10 mg zolpidem per night in 74.8%, 5 mg in 19.8%, 20 mg in 2.4% and > 20 mg in 10 cases. Most physicians (87.6%) rated the efficacy of zolpidem as 'very good' or 'good'. One hundred and eighty-two (1.1%) of the 16 944 patients reported 268 adverse events (one adverse event in 113 cases, two adverse events in 53 cases and more than two adverse events in 16 cases). One hundred and eighteen (64.8%) of these patients (0.006% of all participating patients) discontinued treatment because of adverse events. Nausea (n = 36), dizziness (n = 35), malaise (n = 23), nightmares (n = 20), agitation (n = 19), and headache (n = 18) were the most common adverse events. There was one serious adverse reaction in a 48-year-old women who developed paranoid symptoms during the documentation phase. No life-threatening adverse event occurred. The adverse event profile reflected the pharmacological properties of zolpidem and underlined the cumulative good experience with the drug internationally. PMID- 9727727 TI - One year real world prospective follow-up study of a major depressive episode of patients treated with paroxetine and pindolol or paroxetine for 6 weeks. AB - The objective of our study was an assessment of outcome in a 1 year 'real world', prospective follow-up study of patients who were treated with paroxetine 20 mg plus pindolol 7.5 mg or paroxetine 20 mg plus placebo for 6 weeks. Eighty patients recruited to a double-blind placebo-controlled randomized 6 weeks study of paroxetine and pindolol were followed up for 6 months and interviewed after 1 year. A Kaplan-Meier survival analysis of the patients who relapsed having taken paroxetine and pindolol, compared with those who relapsed having taken paroxetine and placebo, shows that the pindolol group had a better clinical outcome. After 1 year patients who had responded to medication by 2 weeks had done better than the rest and they had been more compliant with medication for 6 months. There are indications that the earlier a patient responds to medication, the better the long-term outcome. PMID- 9727728 TI - Cost effectiveness study of a year follow-up of selective serotonin reuptake inhibitor (SSRI) and augmentor combination compared with SSRI and placebo. AB - We describe a method for evaluating the value of increased cost of pharmacological augmentation that, taken for 6 weeks, accelerates the action of an antidepressant. We test the hypothesis that, if onset of action is taken into account, any added direct costs of the augmenting agent are offset by longer term cost effectiveness. Data to illustrate the method were based on a double-blind randomized placebo controlled study, in which 80 patients originally took part. Patients received the selective serotonin reuptake inhibitor (SSRI) antidepressant paroxetine and an augmenting agent (pindolol) or placebo. After 6 weeks, patients were offered SSRI alone on an open label basis for up to 6 months. At that point they were discharged to their general practitioner or local psychiatric services and subsequently assessed by us at one year. We have used techniques of decision analysis, cost effectiveness and cost benefit and have included a sensitivity analysis. The direct costs over one year of SSRI and augmenting agent, if taking the acceleration effect into account, represented greater cost effectiveness than the SSRI antidepressant alone. The cost effectiveness analysis was positive in both cases. We conclude that the direct costs of treatment are higher than those of previous calculated with SSRIs; but the rate of onset must be taken into account. The application of the model appears valid and useful, and may be used as part of the evaluation of other augmentation regimes. PMID- 9727729 TI - Effects of clorazepate, diazepam, and oxazepam on a laboratory measurement of aggression in men. AB - The effects of three benzodiazepines on human aggressive behavior were examined in 44 medically healthy men. Volunteers were administered either placebo, 10 mg diazepam, 15 mg chlorazepate, or 50 mg oxazepam orally using double-blind procedures. Approximately 90 min after drug ingestion, participants were given the opportunity to administer electric shocks to an increasingly provocative fictitious opponent during a competitive reaction-time task. Aggression was defined as the level of shock the participant was willing to administer to the opponent. Results support the notion that diazepam (but not all benzodiazepines) can elicit aggressive behavior under controlled, laboratory conditions. Implications regarding the clinical use of various benzodiazepines for the tranquilization of potentially assaultive patients are discussed. PMID- 9727730 TI - Clonazepam-induced maniacal reaction in a patient with bipolar disorder. PMID- 9727731 TI - Hemochromatosis: genetics helps to define a multifactorial disease. AB - Hereditary hemochromatosis (HH) is a common autosomal recessive disorder that can result in iron overload and a wide range of clinical complications, including hepatic cirrhosis, diabetes mellitus, hypopituitarism, hypogonadism, arthritis, and cardiomyopathy. People with HH can be detected at an asymptomatic stage of the disease by abnormalities in serum iron measures. Early detection is desirable, because periodic phlebotomy provides effective treatment for iron overload and may prevent complications of the disorder. The natural history of HH is poorly understood, however, and the proportion of people detected by screening who will develop serious complications of HH is unknown. The genetics of HH may help to resolve these questions. The gene, HFE, and two mutations, C282Y and H63D, have been identified: the C282Y mutation has a higher penetrance than the H63D mutation, and appears to result in a greater loss of HFE protein function. Most people with HH are C282Y homozygotes, a small proportion are compound heterozygotes or H63D homozygotes, and some have no identifiable HFE mutation or are HFE heterozygotes, suggesting that additional mutations associated with HH are yet to be found. Gender and environmental agents, such as alcohol and dietary iron, influence phenotypic expression of HH. The severity of HH is thus determined by an interaction between genotype and modifying factors. HFE mutations also appear to increase the likelihood of iron overload in inherited anemias and to promote the clinical manifestations of porphyria cutanea tarda. HH is an important paradigm for medical genetics because it offers an opportunity to explore the complexity of gene gene and gene environment interactions. PMID- 9727732 TI - Recent progress in the molecular genetics of congenital heart defects. AB - Congenital heart defects (CHD) constitute the single most common anatomic class of birth defects and are a major cause of infant mortality. Correlation of normal and pathological embryology/anatomy has led to the formulation of mechanistic models, but there is limited understanding of the genetic basis for the inferred embryological processes. Most evidence points to extensive etiologic heterogeneity and a re-evaluation of simple multifactorial models is required. The recent identification of several genes responsible for congenital heart defects in the context of more complex clinical disorders provides significant entry points for the genetic analysis of human heart development. The association of aneusomies (particularly microdeletion syndromes) with specific cardiac lesions provides further strong support for mechanistic classification. Studies in the mouse are laying the groundwork for a comprehensive genetic model of cardiac organogenesis. Nevertheless, the basis for the large majority of CHD, especially isolated defects, remains obscure. Dissection of the genetic components of CHD is one of the greatest challenges in medical genetics for the coming decades. PMID- 9727733 TI - New insights into the genetics of lissencephaly. PMID- 9727734 TI - An unexpected cause for combined deficiency of coagulation factors V and VIII. PMID- 9727735 TI - Spinal muscular atrophy: a disease of altered RNA metabolism? PMID- 9727736 TI - Linkage analysis in a large Spanish family with X-linked retinitis pigmentosa: phenotype-genotype correlation. AB - X-linked retinitis pigmentosa (XLRP) accounts for 10-25% of RP families and causes the most severe form of the disease in terms of onset and progression. Although three different loci (RP3, RP2 and RP15) have been proposed on the short arm of the X-chromosome by linkage analysis, RP3 represents the disease locus in the majority of XLRP families. The identification of female carriers of X-linked RP is important for genetic counselling. The presence of fundus and electroretinogram (ERG) abnormalities have been reported to be as high as 87 and 90%, respectively. However, in clinical practice it has not always been possible to know the carrier state of females at risk. Thirty-five members of a Spanish family with X-linked RP were evaluated by linkage analysis using nine polymorphic markers (CYBB, DXS1110, M6, DXS6679, DXS1068, DXS1058, MAOA, MAOB and DXS6849) that map to the X-chromosome region Xp21.1 to Xp11.3, in an attempt to determine the carrier state of these females at risk. It was possible to establish that a RP3 mutation is, most likely, segregating in this family. PMID- 9727737 TI - Association of long variants of the dopamine D4 receptor exon 3 repeat polymorphism with Parkinson's disease. AB - The dopamine D4 receptor (D4DR) has a highly polymorphic region in the third exon which has been associated with novelty seeking (NS) behavior. Due to the central position of dopamine and the documented low NS in Parkinson's disease (PD), the frequency of the exon 3 variants of D4DR in 95 PD patients and 47 controls was investigated. A significantly higher frequency of exon 3 alleles with six or more repeat units was found in the PD group (p = 0.039). This provides evidence that some forms of the highly polymorphic D4DR may represent a genetic susceptibility factor for PD. PMID- 9727738 TI - Low frequency of RET mutations in Hirschsprung disease in Sweden. AB - Hirschsprung disease is a congenital malformation, where absence of intramural ganglia in the hindgut results in a defect in the coordination of peristaltic movement. This leads to ileus in the newborn or, more often, constipation in children and adults. The disease affects one in 5000 live births. Siblings of affected cases are at an increased risk (4%) of developing the disease. Among cases. males are affected more often than females. The first major susceptibility gene for Hirschsprung disease is the RET proto-oncogene on 10q11.2. Germline RET mutations in Hirschsprung disease are mainly inactivating, and have been reported to account for up to 20 and 50% of sporadic and familial cases, respectively. We have screened Swedish population-based samples from 62 sporadic cases and seven familial cases of Hirschsprung disease with single strand conformation polymorphism (SSCP), and found five mutations. PMID- 9727739 TI - Analysis of an interstitial deletion in a patient with Kallmann syndrome, X linked ichthyosis and mental retardation. AB - Contiguous gene syndromes are an interesting clinical phenomenon, resulting from interstitial or terminal deletions of several adjacent genes. The phenotype results in a combination of two or more monogenic disorders and relates clinical findings to corresponding genotypes. We present the case of a male patient with Kallmann syndrome (KS), X-linked ichthyosis (XLI) and X-linked mental retardation (MRX). He was referred at the age of 15.4 years for delayed puberty and obesity. He had a previous history of pyloric stenosis, bilateral orchidopexy and surgical correction of a pes equinovarus adductus. On physical examination, generalised ichthyosis and hypoplastic external genitalia were found. KS was evident with hypogonadotropic hypogonadism, hyposmia and a hypoplastic anlage of the olfactory tract in magnetic resonance imaging. Lipoprotein electrophoresis, and lack of steroid sulfatase and arylsulfatase-C activity in leucocytes confirmed XLI. DNA investigation established an interstitial deletion in Xp22.3 involving the Kallmann (KAL) gene, the steroid sulfatase (STS) gene and a putative mental retardation locus (MRX). The novel MRX locus maps to a 1-Mb region between DXS1060 and GS1. PMID- 9727740 TI - Female external genitalia, absent uterus, and probable agonadism in a 46,XY infant with bilateral upper amelia. AB - This report describes a 46,XY phenotypic female infant with absent uterus, probable agonadism, and bilateral upper amelia. The constellation of anomalies is similar to that of the patient described by Temocin et al. (Acta Paediatr Jpn 1997: 39: 631-633), and may suggest a developmental link between genital region and upper limbs. PMID- 9727741 TI - Phenotypic and genotypic variability in monozygotic triplets with Turner syndrome. AB - Turner syndrome (TS) is a common disorder (1/2500 and 1/5000 female births) which is diagnosed at birth in approximately 20% of patients and during childhood (usually due to growth retardation) or later, (due to lack of pubertal development) for the remaining patients. Here we present a cytogenetic and molecular analysis of three monozygotic sisters. The diagnosis of TS was done for one of them (patient 1) who presented with a typical Turner phenotype. A first karyotype was established as normal and a second karyotype (carried out on 200 cells) revealed a 45,X/46,XX mosaicism with 6% of cells with a 45,X karyotype. Lymphocyte karyotype analysis showed the same mosaicism pattern for the two other sisters, one of them exhibiting only a mild (patient 2) and the other no clinical features of Turner syndrome (patient 3). Karyotype analysis was this time conducted on fibroblasts and showed that the 45,X/46,XX mosaicism pattern correlated with the clinical phenotype with 99, 43 and 3% of 45,X cells in patients 1, 2, and 3, respectively. These data suggest that different tissues other than lymphocytes should be subjected to a karyotype analysis when the observed genotype does not correlate with the clinical phenotype. PMID- 9727742 TI - A case of Prader-Willi syndrome arising as a result of familial unbalanced translocation t(11;15)(q25;q13). AB - We report on a case of Prader-Willi syndrome (PWS) with a true reciprocal unbalanced translocation, 45,XX,-15,der(11)t(11;15)pat. The proposita was diagnosed clinically as having severe PWS. Molecular studies revealed loss of the paternal methylation pattern at locus D15S63 and a deletion encompassing the loci from at least D15S10 to D15S97 of paternal chromosome 15. FISH studies confirmed the deletion of 15q11q13 region and the presence of two telomeres on all chromosomes. The proposita's father, the father's sister and their mother are all carriers of the same balanced translocation t(11;15)(q25;q13). By genomic imprinting we would expect that if the father's sister were to give birth to a child with the same unbalanced translocation as the proband, it would be affected by Angelman syndrome. To date, a similar familial unbalanced translocation due to loss of the small chromosome15 derivative has not been described. PMID- 9727743 TI - De novo direct duplication 2 (p12-->p21) with paternally inherited pericentric inversion 2p11.2 2q12.2. AB - We report a 4-year-old girl with a previously undescribed de novo duplication of 2p12->2p21 on the same homologue as a paternally inherited pericentric inversion of region 2p11.2-->2q12.2, resulting in dysmorphic features, cardiac abnormality, cleft palate, respiratory problems, severe growth retardation and developmental delay. This case raises an important question--did the paternal pericentric inversion influence the occurrence of the de novo duplication? PMID- 9727744 TI - Functional mosaic trisomy of 1q12-->1q21 resulting from X-autosome insertion translocation with random inactivation. AB - Cytogenetic studies of a 16-year-old female with behaviour and learning problems revealed that one X chromosome had additional material inserted at Xq21. Fluorescence in situ hybridization (FISH) analysis showed that the inserted segment contained heterochromatin and adjacent euchromatin of chromosome 1 origin. The karyotype of this patient was established as: 46,X,der(X)ins(X;?)(q21;?).ish der(X) ins(X;1)(q21;q12q21)(wcp1+). Chromosome replication studies demonstrated a random pattern of X inactivation, suggesting that the inserted material may be too 'small' to skew lyonization. The consequences of this abnormal X chromosome in relation to the clinical phenotype are discussed. PMID- 9727745 TI - Familial hypercholesterolemia: potential diagnostic value of mutation screening in a pediatric population of South Africa. AB - Three founder-related low-density lipoprotein receptor (LDLR) gene mutations, D154N, D206E and V408M, cause familial hypercholesterolemia (FH) in approximately 90% of South African Afrikaners. Two hundred and twenty-one South African children, from 85 affected families, were screened for the specific mutation identified previously in the index case. Sixty boys and 56 girls were heterozygous for mutation D154N (FH3), D206E (FH1) or V408M (FH2). Total and LDL cholesterol (LDLC) levels were similar among the children heterozygous for the three founder mutations, and mean values were significantly higher compared to those without a known mutation (p < 0.0001). Plasma cholesterol levels overlapped considerably between the different groups, suggesting that modifiable lifestyle factors remain important in children with FH. This study demonstrates the potential diagnostic value of mutation screening in a pediatric population with an enrichment of particular gene mutations. PMID- 9727746 TI - Mutation screening of the LDLR gene and ApoB gene in patients with a phenotype of familial hypercholesterolemia and normal values in a functional LDL receptor/apolipoprotein B assay. AB - Mutations in the LDL receptor (LDLR) or the apolipoprotein B-100 genes causing familial hypercholesterolemia (FH) and familial defective apolipoprotein B-100 (FDB), two of the most frequent inherited diseases, are the underlying genetic defects in a small proportion of patients suffering from premature atherosclerotic heart disease. Consequently, secure diagnostic tools for these conditions allowing early preventive measures are needed. Since clinical and biochemical diagnosis often is inaccurate, assays analyzing patient LDLR function and LDL affinity have been established. These assays are, however, not able clearly to differentiate between suspected FH/FDB samples and normal controls. To evaluate if this may be caused by other hitherto undescribed genetic defects or to failure of the functional assays, we undertook denaturing gradient gel electrophoresis based mutation screening of the LDLR gene and the codon 3456 3553 region of the apolipoprotein B gene in six French FH/FDB patients with normal outcomes on functional assays. In all six patients, pathogenic LDLR mutations were found, including three previously undescribed mutations, suggesting that failure of the functional assays explains the normal results found in some phenotypic FH/FDB patients and illustrating the need for DNA based screening techniques for routine genetic diagnosis in FH/FDB. PMID- 9727747 TI - Autosomal dominant inheritance of adducted thumbs and other digital anomalies. AB - Isolated adducted thumbs is an uncommon malformation that occurs sporadically in the majority of cases although some affected families have been reported. Previously, autosomal dominant inheritance was suggested in two familial cases, but this mode of inheritance has not been confirmed. Here we describe a family with adducted thumbs and other digital anomalies in which seven members (six females and one male) are affected in three consecutive generations. Additionally, the patients showed mild abnormalities of fingers 2nd-4th bilaterally and hypoplasia of the middle phalanx of the 5th fingers. This family represents an autosomal dominant condition that apparently has not been previously reported. PMID- 9727748 TI - Marden-Walker syndrome versus isolated distal arthrogryposis: evidence that both conditions may be variable manifestations of the same mutated gene. AB - In this report we present evidence that Marden-Walker syndrome and isolated distal arthrogryposis may be variable manifestations of the same entity. We describe the clinical and pathological findings in two affected siblings, the first two children of normal, non-consanguineous parents. The first child, a female, presented a typical Marden-Walker syndrome with Dandy Walker type CNS malformation, corpus callosum hypoplasia and enlarged ventricles. In the second pregnancy, echographic examination revealed joint contractures of the hands and feet. Fetopathological examination revealed a normocephalic male fetus with severe distal arthrogryposis. There was no facial dysmorphism and pathological examination of the brain, the spinal cord and muscle was normal. PMID- 9727749 TI - Infectious complications of propionic acidemia in Saudia Arabia. AB - A retrospective study of 38 patients with propionic acidemia indicates a high frequency of infections; affecting 80% of such patients. The Saudi Arabian population studied is a product of consanguineous marriages, and presents with a severe phenotype. Most microorganisms implicated are unusual, which suggests an underlying immune deficiency. These frequent infections occur despite aggressive treatment with appropriate diets, carnitine and during acute episodes of the disease with metronidazole, which suggests a global effect of the disease on T and B lymphocytes as well as on the bone marrow cells. Any patient with propionic acidemia should be closely followed up for an intercurrent infection in association with acute metabolic decompensation. PMID- 9727750 TI - SKALP/elafin gene polymorphisms are not associated with pustular forms of psoriasis. AB - Psoriasis is a multifactorial skin disease characterised by epidermal abnormalities and infiltration by lymphocytes and polymorphonuclear leukocytes (PMN). Skin-derived antileukoproteinase (SKALP), also known as elafin, is a potent inhibitor of human leukocyte elastase and proteinase 3, two PMN-derived proteinases implicated in tissue destruction and leukocyte migration. We have shown that, at least at the protein level, SKALP is significantly decreased in lesional skin of patients with pustular psoriasis compared with plaque-type psoriasis. This finding raised the possibility that SKALP could be one of the candidate genes for pustular forms of psoriasis. We therefore performed single strand conformation polymorphism (SSCP) analysis on the SKALP gene to screen for mutations/polymorphisms in the exons of 30 patients with plaque-type psoriasis, 15 patients with pustular psoriasis and 48 healthy controls. In exon 1 a polymorphism was detected at position +43 relative to the translation start site, resulting in a substitution of threonine for alanine in the signal peptide. In the promoter region a dinucleotide repeat polymorphism was identified. Both polymorphisms were not associated with pustular psoriasis, or psoriasis in general. Our data indicate that the decrease in SKALP activity in pustular psoriasis is not caused by mutations in the coding region of the gene, and that there is no allelic association between pustular psoriasis and SKALP gene polymorphisms. PMID- 9727751 TI - Polymorphic exonic CAG microsatellites in the gene amplified in breast cancer (AIB1 gene). PMID- 9727752 TI - SacI identifies a biallelic polymorphism in the coding sequence of the gamma subunit of the epithelial sodium channel (ENaC): a candidate gene for hypertension. PMID- 9727754 TI - Phenotypic spectrum of tetrasomy 12p and prenatal counseling: potential underestimation of severity. PMID- 9727753 TI - Apolipoprotein E and A-IV polymorphisms in the Estonian population. PMID- 9727755 TI - The use of a specific clinical history in counselling a family with the balanced translocation 46,XY,t(4;12)(p15.2;q21.3): viable offspring with partial monosomy 4p and trisomy 12q. PMID- 9727756 TI - Further insights into early acquisition of Pseudomonas aeruginosa. PMID- 9727758 TI - Pseudomonas aeruginosa and Burkholderia cepacia infection in cystic fibrosis patients treated in Toronto and Copenhagen. AB - Differences in the course of pulmonary disease in cystic fibrosis (CF) may be altered by different treatment strategies in different CF centers. The Copenhagen clinic uses scheduled, regular and very aggressive treatment of lung infection. The Toronto clinic treats pulmonary infection with oral, inhaled, or intravenous antibiotics, and has emphasized aggressive nutritional therapy. This study compared the clinical status of CF patients treated in the two centers (Toronto, Canada, n=302, and Copenhagen, Denmark, n=214) using a cross-sectional design in terms of Pseudomonas aeruginosa (PA) and Burkholderia cepacia (BC) lung infections, pulmonary function, and levels of PA and BC precipitating antibodies (precipitins). Median ages were similar, but the age distribution was significantly different, with a higher proportion of patients under 10 and > or = 25 years in Toronto, and higher proportion of patients 11-24 years of age in Copenhagen. A higher number of female patients was observed in Copenhagen than in Toronto. Seventy-nine percent of Copenhagen patients, and 52% of Toronto patients were deltaF508 homozygous. Of all the patients, 20.1% of Copenhagen patients and 38% of Toronto patients were deltaF508 heterozygous. Ten percent of Toronto patients had two uncommon mutations. Pulmonary function and nutritional status in both groups were similar despite varying treatment strategies. The prevalence of PA was lower in Danish children and higher in Danish adults than in Canada. These differences are probably due to cohort isolation, which was introduced in Copenhagen in 1981. The prevalence of BC was higher in Toronto than in Copenhagen patients at all ages. In both centers, the number of PA and BC precipitins increased with age in patients chronically infected with PA and BC, respectively, and the number of both PA and BC precipitins rose with declining lung function. This study suggests that the clinic populations had similar pulmonary and nutritional statuses despite differing clinic antibiotic treatment strategies. Microbial colonization seemed to differ, at least in part, because of differences in cohort isolation strategies. Early, aggressive anti-pseudomonal chemotherapy may have reduced pseudomonal colonization among younger patients in Copenhagen. Future studies will be required to assess the impact of this on this cohort's outcome. PMID- 9727757 TI - Comprehensive analysis of risk factors for acquisition of Pseudomonas aeruginosa in young children with cystic fibrosis. AB - The objective of this study was to identify risk factors of significance for acquisition of Pseudomonas aeruginosa by children with cystic fibrosis (CF). Our working hypothesis is that exposure of infants and young children with CF to older, infected patients increases their risk for acquiring this organism. A special opportunity arose to study this question in detail, as we have been performing a randomized clinical trial of neonatal screening for CF throughout the state of Wisconsin during the period of 1985-1994. Patients were selected for this study based on either early identification through screening or diagnosis by standard methods. A longitudinal protocol employed at Wisconsin's two CF Centers includes routine cultures of respiratory secretions and collection of clinical, demographic, and activity information on patients and their families. Previous observations in our trial revealed that one center at an old hospital in an urban location showed a significantly shorter time to acquisition of P. aeruginosa for CF patients followed there. To study the center effect further, we performed statistical analyses using survival curves and stepwise regression analysis of all life history covariates available. The results of these analyses showed that the statistically significant correlations involve the following risk factors: 1) center and old hospital (r=0.42); 2) center and original physician (r=0.61); 3) center and exposure to pseudomonas-positive patients (r=0.29); and 4) population density and urban location (r=0.49). The final statistical model demonstrated that increased risk due to aerosol use (odds ratio=3.45, P=0.014) and a protective effect associated with education of the mother (odds ratio=0.81, P=0.024) were the most significant factors for acquisition of P. aeruginosa. The previously observed center effect was confined to the 1985-1990 interval at the old hospital (odds ratio=4.43, P < 0.001). We conclude that multiple factors are involved in increasing the risk of young children with CF to acquire P. aeruginosa, and that the observed center effect can best be explained by a combination of factors. These results suggest that facilities and methods used to care for young children with CF can significantly influence their likelihood of acquiring pseudomonas in the respiratory tract. PMID- 9727759 TI - Airway inflammation after treatment with aerosolized deoxyribonuclease in cystic fibrosis. AB - Recombinant human deoxyribonuclease (rhDNase) has been shown to reduce sputum viscoelasticity and to improve lung function in patients with cystic fibrosis (CF). The aim of this study was to determine whether airway inflammation would decrease after administration of rhDNase. Twenty patients with CF and chronic suppurative lung disease inhaled 2.5 mg of rhDNase daily for 1 month. Before and after the 1-month trial, lung function was measured and sputum was obtained, either after spontaneous expectoration or after sputum induction with hypertonic saline. Sputum total cell and differential counts were measured using techniques previously described. The mean age of the patients was 16.8 years (range, 6.7 27.5). After 1 month of rhDNase, mean FEV1 increased from a baseline of 62.3% predicted to 70.8% (P= 0.02, paired t test); and FVC increased from 74.4% to 83.9% predicted (P=0.007). No significant differences were found in sputum cytology before or after rhDNase (median total cell counts 16.0 x 10(6)/ml vs. 19.3 x 10(6)/ml, P=0.68). Thirteen patients had a 10% or greater increase in FEV1 after rhDNase (responders). Initial lung function was less in responders than in nonresponders (53.5% vs. 78.6%, P=0.007). There was no significant change in total cell count and neutrophil count after rhDNase in either responders or nonresponders. We conclude that airway inflammation, as measured by total cell counts in sputum, was a prominent feature in cystic fibrosis, and neutrophils were the dominant inflammatory cells. Although the administration of rhDNase resulted in significant improvements in FEV1, there was no evidence of accompanying changes in airway inflammation. PMID- 9727760 TI - Time to publication as full reports of abstracts of randomized controlled trials in cystic fibrosis. AB - OBJECTIVES: To determine 1) what proportion of abstracts of randomized controlled trials (RCTs) presented at international conferences on cystic fibrosis (CF) are published as full reports, 2) time to publication, and 3) factors that might delay or prevent publication. METHODS: At the end of 1995, the Cochrane CF Group's register of RCTs contained 199 abstracts describing 180 RCTs. Abstracts were identified by handsearching 44 abstract books of three international CF conferences over a 30-year period. We searched the register for subsequent full reports of these RCTs and used survival analysis to investigate time to publication. Using the log-rank test, we examined 1) whether there is a difference in time to publication between reports where the investigators concluded that the test treatment was as effective as or better than the control treatment, and reports where it was not, and 2) whether there is a difference in time to publication according to sample size. RESULTS: Thirty-two percent of the 178 abstracts analyzed were subsequently published in full. Survival analysis indicated that the proportions published before 12 months, 2 years, and 5 years were 8.1%, 29%, and 40% respectively. No difference in time to publication was identified when the abstracts were stratified according to conclusions or sample size; no significant association (P > 0.05) existed between time to publication and both sample size and conclusions together. CONCLUSION: Only a small proportion of abstracts of RCTs presented at international conferences on CF are followed by full publication, and usually only after several years. Therefore, many potentially valuable studies do not reach a wide audience. However, we found no consistent factors which might delay or prevent publication. PMID- 9727761 TI - Prevalence of asthma or respiratory symptoms among children attending primary schools in Paris. AB - The objectives of this study were to determine the prevalence of chronic respiratory symptoms and asthma in 8- to 9-year-old children in Paris, and to analyze their medical management. This cross-sectional study was carried out in 1994 on a randomized sample of 3,756 pupils attending Paris public primary schools. The response rate by parents to an initial standardized self administered questionnaire was 94.8%. This questionnaire identified 601 children (17%) as having recurrent respiratory symptoms. Of these children, 555 (92.3%) agreed to participate in a follow-up survey that evaluated their medical management; these children were examined by 73 school doctors of the city of Paris. Prevalence of parent-reported doctor-diagnosed asthma was 6.1%. In addition to these 211 children with asthma, 344 other children had recurrent respiratory symptoms: 120 children were "wheezers," and the remaining 224 children were "coughers." Among "chesty" pupils not identified as asthmatics, physical education teachers reported exercise-induced cough or respiratory discomfort in 13.7%, and nearly 14% had a peak expiratory flow 20% lower than the predicted values for age and height. In children identified as asthmatic, 25.3% were not under medical supervision, 55.5% had never performed lung function tests, 63.7% did not receive any prophylactic treatment, and 59.7% were receiving no treatment. Bronchodilator prophylactic medication before exercise was used by only 7% of asthmatics, although physical training teachers noted chest discomfort on exercise in 30.4% of these pupils. These results confirm that children with asthma and participating in this study were less than optimally investigated, were underdiagnosed and undertreated, and their medical management was not optimal. In addition to its epidemiologic value, the study has helped Paris school doctors to advise parents to refer their children to their general practitioner when asthma was suspected or undertreated. PMID- 9727762 TI - One-year follow-up of young children hospitalized for wheezing: the influence of early anti-inflammatory therapy and risk factors for subsequent wheezing and asthma. AB - We investigated the 1-year outcome of children hospitalized for wheezing, paying special attention to the effect of early anti-inflammatory therapy. In addition, we identified risk factors for recurrent wheezing and asthma. Eighty-eight children under 2 years old treated in the hospital for wheezing were followed for 1 year. Nebulized anti-inflammatory therapy was given for 16 weeks: 31 patients received budesonide, 29 patients cromolyn sodium, and 28 control patients received no therapy. The number of subsequent physician-diagnosed wheezing episodes was recorded. Four months of anti-inflammatory therapy did not significantly decrease the occurrence of asthma 1 year later; 45% of patients in the cromolyn group, 42% in the budesonide group, and 61% in the control group had asthma, defined as at least two bronchial obstruction episodes during the 1-year period after the original hospitalization for wheezing. An age over 12 months at the time of the initial bronchial obstructing episode [P=0.009, risk ratio (RR)=5.4, 95% confidence interval (CI)=1.53-19.31], failure to identify a viral cause (P=0.0003, RR=12.0, CI=3.16-45.40), history of wheezing (P=0.02, RR=14.6, CI=1.59-132.10), the presence of atopy (P=0.01, RR=5.3, CI=1.47-19.21), a family history of atopy (P=0.03, RR=3.6, CI =1.15-11.12), and serum eosinophil cationic protein (ECP) > or = 16 microg/L (P=0.005) were significant risk factors for asthma. We conclude that early anti-inflammatory therapy for 4 months does not significantly decrease the occurrence of asthma during the period of 1 year following hospitalization for the original episode of wheezing. Young children requiring hospital admission for wheezing during a respiratory tract infection are at increased risk of having subsequent asthma if they have wheezed previously, if they have atopy or a family history of atopy, if they have elevated serum ECP, if they are over 12 months of age at the original bronchial obstructive episode, and especially when viral studies are negative. PMID- 9727763 TI - Indoor environmental risk factors and childhood asthma: a case-control study in a subtropical area. AB - This study examined the relation between indoor environmental factors and childhood asthma in a subtropical area. A hospital-based case-control study was performed in Kaohsiung, Taiwan, between July of 1995 and June of 1996. Eighty-six children seen in the out-patient clinic of our university hospital and who had a first-time diagnosis of asthma made by a pediatrician were the test group; 86 control subjects were selected from children attending the Childhood Orthopaedic Clinic in the same hospital and who had no previous diagnosis of asthma or asthma symptoms and no history of physician confirmed atopic diseases. The control subjects were matched with test case children on the basis of gender and age. Information was obtained from parents using a structured questionnaire. Of the many indoor environmental factors included in this study, only home dampness showed an association with asthma (adjusted odds ratio=1.77; 95% confidence intervals, 1.24-2.53). We conclude that dampness in the home is a new public health risk factor related to asthma in subtropical areas. PMID- 9727764 TI - Ventilation index and outcome in children with acute respiratory distress syndrome. AB - The purpose of this investigation was to determine the predictive value of the ventilation index (VI) in children with acute respiratory distress syndrome (ARDS). We performed a 10-year retrospective chart review of children who were admitted to the Pediatric Intensive Care Unit with a diagnosis of ARDS. Acute respiratory distress syndrome was defined as acute onset of diffuse, bilateral pulmonary infiltrates of noncardiac origin, and severe hypoxemia, defined as the ratio of the arterial partial pressure of oxygen to the fraction of inspired oxygen of <200 and a positive end expiratory pressure of 6 cmH2O or greater. Records of daily arterial blood gas results and ventilator settings were reviewed, and the ventilation index (VI=partial pressure of arterial CO2 x peak airway pressure x respiratory rate/1,000) was calculated each time the measurements were made. These values were correlated with outcome (survival or nonsurvival). The VI was not different at the time of diagnosis of ARDS in the patients who lived, compared with those who subsequently died. However, by 3 to 5 days after study entry, the VI of nonsurvivors was significantly higher than for survivors (P < 0.05). The VI for survivors remained between 30 and 35 throughout the study period, whereas the VI of nonsurvivors continued to increase with time. A VI of >65 predicted death with a specificity and positive predictive value of >90% on days 3 through 9. We conclude that the VI provides a reliable prognostic marker in children with ARDS, and its increase above 65 indicates a need for orderly intervention with alternative modalities of care. PMID- 9727765 TI - Comparison of rapid bolus instillation with simplified slow administration of surfactant in lung lavaged rats. AB - The aim of this study was to compare the effects of modified porcine surfactant (Curosurf) given either by a simplified slow delivery technique or by the standard bolus method, on pulmonary gas exchange, lung mechanics, and surfactant distribution in rats with respiratory failure produced by lung lavage. Twelve rats with respiratory failure induced by lung lavage received 200 mg x kg(-1) body weight (b.w.) of tagged porcine surfactant, either by the standard bolus delivery technique or by a simplified 1-min intratracheal infusion method, not requiring interruption of mechanical ventilation. Cardiovascular parameters, arterial blood gases, and pulmonary mechanics were measured repeatedly. Surfactant distribution was also measured by dye-tagged microbead spheres. After surfactant administration, there were no overall major differences between groups in mean heart rate, blood pressure, arterial blood gases, dynamic lung compliance, respiratory system resistance, and pulmonary distribution of exogenous surfactant. However, after 180 min pulmonary gas exchange was better and compliance higher in the bolus than the 1-min infusion group. A transient decrease in blood pressure and heart rate was observed in the bolus group; this side effect was not seen in animals treated with the simplified 1-min infusion method. We conclude that in rats subjected to lung lavage, the infusion of porcine surfactant by a simplified 1-min procedure produced similar short-term effects compared to the same dose of surfactant given by the bolus method. We speculate that tracheal bolus dosing is highly effective and might be the preferable delivery method for porcine surfactant. Dosing by the simplified method described appears less effective, but since no significant differences were observed, and since it produced less acute adverse effects, it could be used when clinical circumstances preclude rapid delivery. PMID- 9727766 TI - Chronic anaerobic pneumonitis in a seven-year-old girl. AB - A 7-year-old girl was referred for evaluation of chronic pulmonary disease associated with nasal symptoms of 4 years duration for which she had received frequent courses of antibiotics. Serial chest roentgenograms over a period of 2 years revealed a nonhomogeneous opacity in the right lower lung zone for which she had received 18 months of antituberculous therapy without relief. Evaluation of the patient led to the diagnosis of chronic anaerobic pneumonitis, a rare clinical entity in children. In addition, the patient also had bronchial asthma and chronic rhinitis. Therapy with oral phenoxymethylpenicillin and metronidazole for 6 weeks along with appropriate antiasthma medications abolished her symptoms and resulted in roentgenologic clearance. PMID- 9727768 TI - Of shadows and wind. PMID- 9727767 TI - Right-sided pulmonary aplasia: longitudinal lung function studies in two cases and comparison to results from term healthy neonates. AB - Agenesis of the right lung was diagnosed prenatally in two neonates born at 36 and 37 weeks, respectively. Computed tomographic scans and magnetic resonance imaging indicated that both cases had a Type 2 pulmonary agenesis, which was confirmed later by bronchoscopy. Both patients were clinically stable during the neonatal period. Serial pulmonary function tests revealed a decrease in specific respiratory system compliance (sCrs) in both neonates and a marked discrepancy between functional residual capacity measured by the nitrogen washout technique (FRCN2) and by plethysmography (FRCpleth) on follow-up. Early decrease of respiratory system compliance (Crs) and increase of respiratory system resistance (Rrs) in one infant preceded the onset of tracheal stenosis, which remained asymptomatic until the age of 8 weeks, when the infant developed acute respiratory failure requiring intubation and mechanical ventilation with high airway pressures. Aortopexy, implantation of a tissue expander into the right hemithorax, and laser ablation of fibrotic tissue at the site of tracheal stenosis were performed to achieve successful extubation. The second infant remained asymptomatic. Values for lung mechanics and volumes for both infants with pulmonary aplasia were as follows: Crs, 3.43 and 10.60 mL x kP(-1) x kg(-1); sCrs, 0.23 and 1.28 kpa(-1); Rrs, 11.1 and 7.4 kpa x s x L(-1); FRCN2, 14.9 and 10.2 mL x kg(-1); FRCpleth, 28.2 and 25.8 mL x kg(-1); FRCN2: FRCpleth ratio, 0.56 and 0.54 for patients 1 and 2, respectively. These values differed considerably from results of a control group of nine term healthy neonates (Crs, 10.0+/-1.8 mL x kPa(-1) x kg(-1); sCrs, 0.43+/-0.08 kpa(-1); Rrs, 5.10+/-0.55 kpa x s x L(-1); FRCN2, 24.0+/-2.5 mL x kg(-1); FRCpleth, 31.1+/-6.0 mL x kg(-1); FRCN2:FRCpleth ratio, 0.78+/-0.10). In conclusion, serial assessment of lung mechanics and pulmonary gas volumes detects airway obstruction early in neonates with unilateral lung agenesis. Bronchoscopy is recommended. Along with conventional surgical procedures, an expandable implant may improve management or prevent respiratory failure in selected cases. PMID- 9727770 TI - Performance testing new peak flow meters. PMID- 9727769 TI - Safety of endotracheal rh DNAse (Pulmozyme) for treatment of pulmonary atelectasis in mechanically ventilated children. PMID- 9727771 TI - Vasodilators in the treatment of primary pulmonary hypertension. PMID- 9727772 TI - Inhaled nitric oxide as a screening agent for safely identifying responders to oral calcium-channel blockers in primary pulmonary hypertension. AB - In a subset of patients with primary pulmonary hypertension (PPH), high doses of oral calcium-channel blockers (CCB) produce a sustained clinical and haemodynamic improvement. However, significant side-effects have been reported during acute testing with CCB. Therefore, to identify accurately patients who may benefit from long-term CCB therapy, there is a need for a safe, potent and short-acting vasodilator. The aim of this study was to compare the acute response to inhaled nitric oxide (NO) and oral high doses of CCB in 33 consecutive patients with PPH. A significant acute vasodilator response was defined by a fall in both mean pulmonary artery pressure and total pulmonary resistance by >20%. Ten patients responded acutely to NO, nine of whom responded acutely to CCB, without any complications. The 23 other patients failed to respond to NO and CCB. In these nonresponders, nine serious adverse events were observed with CCB (38%). There was no clinical or baseline haemodynamic feature predicting acute vasodilator response. Long-term oral treatment with CCB was restricted to the nine acute responders and a sustained clinical and haemodynamic improvement was observed in only six patients. In primary pulmonary hypertension, the acute response rate to high doses of calcium-channel blockers is low (27%). Serious adverse reactions to high doses of calcium-channel blockers during acute testing are frequently observed in nonresponders. It is concluded that nitric oxide may be used as a screening agent for safely identifying patients with primary pulmonary hypertension who respond acutely to calcium-channel blockers and may benefit from long-term treatment with these agents. PMID- 9727773 TI - High incidence of primary pulmonary hypertension associated with appetite suppressants in Belgium. AB - Primary pulmonary hypertension is a rare, progressive and incurable disease, which has been associated with the intake of appetite suppressant drugs. The importance of this association was evaluated in Belgium while this country still had no restriction on the prescription of appetite suppressants. Thirty-five patients with primary pulmonary hypertension and 85 matched controls were recruited over 32 months (1992-1994) in Belgium. Exposure to appetite suppressants was assessed on the basis of hospital records and standardized interview. Twenty-three of the patients had previously taken appetite suppressants, mainly fenfluramines, as compared with only 5 of the controls (66 versus 6%, p<0.0001). Five patients died before the interview, all of them had taken appetite suppressants. In 8 patients the diagnosis of primary pulmonary hypertension was uncertain, 5 of them had taken appetite suppressants. The patients who had been exposed to appetite suppressants tended to be on average more severely ill, and to have a shorter median delay between onset of symptoms and diagnosis. A policy of unrestricted prescription of appetite suppressants may lead to a high incidence of associated primary pulmonary hypertension. Intake of appetite suppressants may accelerate the progression of the disease. PMID- 9727774 TI - Pulmonary hypertensive effects of lung inflation in chronic hypoxia: a study in rats. AB - Lung inflation was compared in isolated perfused lungs of control (C) and chronically hypoxic (CH) rats; in the latter, there is muscularization and loss of compliance in the pulmonary arterial system. During ventilation hypoxia, high alveolar pressure (Palv) elevated the pulmonary artery pressure (Ppa) less in C than in CH rats; Ppa fell during sustained inflation, rose on deflation, and after inflation hypoxic vasoconstriction was attenuated. Opposite changes took place in CH rats; Ppa often rose during inflation, fell on deflation, and after inflation hypoxic vasoconstriction was enhanced. Inflation also increased Ppa more in CH than C rats during air ventilation. Ppa/Palv relations measured during incremental inflation revealed normoxic tone in "extra-alveolar" vessels in both rat groups, which usually increased during hypoxia. In CH, but not C rats, there was also evidence for constriction in "alveolar" vessels during hypoxia. The effects of inflation were not changed by NO synthase blockade in either rat group. Pulmonary hypertensive effects of inflation in chronically hypoxic rats can be attributed to vascular remodelling. PMID- 9727775 TI - Surfactant therapy restores gas exchange in lung injury due to paraquat intoxication in rats. AB - Paraquat is a weed killer which causes often fatal lung damage in humans and other animals. There is evidence that the pulmonary surfactant system is involved in the pathophysiology of respiratory failure after paraquat intoxication and, therefore, the possible therapeutic effect of intratracheal surfactant administration on gas exchange in rats with progressive lung injury induced by paraquat poisoning was studied. In one group of rats, the time course of the development of lung injury due to paraquat intoxication was characterized. In a second group of rats, 72 h after paraquat intoxication, the animals underwent mechanical ventilation and only those animals in which the arterial oxygen tension/inspiratory oxygen fraction (Pa,O2/FI,O2) decreased to below 20 kPa (150 mmHg) received exogenous surfactant (200 mg x kg(-1) body weight). Within 3 days the rats in group 1 developed progressive respiratory failure, demonstrated not only by impaired gas exchange and lung mechanics but also by increased minimal surface tension and increased protein concentration in bronchoalveolar lavage fluid. In group 2, intratracheal surfactant administration increased Pa,O2/FI,O2 significantly within 5 min (14.4+/-2.4 kPa (108+/-18 mmHg)) to (55.2+/-53 kPa (414+/-40 mmHg)) and sustained this level for at least 2 h. It is concluded that intratracheal surfactant administration is a promising approach in the treatment of severe respiratory failure caused by paraquat poisoning. PMID- 9727776 TI - Breathing and pulmonary surfactant function in mice 24 h after ozone exposure. AB - The aim of this study was to determine whether an acute ozone exposure affects breathing, and the ability of pulmonary surfactant to maintain the patency of terminal conducting airways. BALB/c mice were exposed to ozone (1 part per million (ppm)) for 2, 4, 6, and 8 h. They were examined with plethysmography and with bronchoalveolar lavage (BAL) 24 h later. The BAL fluid was analysed for the presence of inflammatory cells and concentrations of proteins and phospholipids. Surfactant in the remaining BAL fluid was concentrated five-times and examined with a capillary surfactometer (CS). The surfactant was then washed with a large volume of saline solution which was removed following centrifugation. Already, after a 2 h ozone exposure, the respiratory frequency increased from 297+/-6 to 386+/-11 breaths x min(-1) (p<0.0001). Pressure amplitude per breath diminished (p<0.001), indicating a reduced tidal volume. A highly significant surfactant dysfunction was observed with the CS (p<0.0001), although phospholipids increased. However, proteins also increased (p<0.0001) and they or other water soluble inhibitors apparently caused the surfactant dysfunction since, when they were removed with a washing procedure, the surfactant's normal ability to maintain patency was restored. The acute ozone exposure affected breathing and caused an airway inflammation. The inflammatory proteins or other water-soluble inhibitors reduced the surfactant's ability to secure airway patency. PMID- 9727777 TI - Uptake and degradation of Curosurf after tracheal administration to newborn and adult rabbits. AB - This paper examines the removal from the airways of Curosurf, a commercial surfactant derived from porcine lungs, administered at pharmacological concentrations to newborn or adult animals. Curosurf was labelled by the addition of radioactive dipalmitoyl phosphatidylcholine (DPPC) and administered intratracheally to newborn and adult rabbits at a dose of 200 mg x kg(-1) body weight. The disappearance of DPPC from the airways and its appearance in alveolar macrophages, lung parenchyma, lamellar bodies, serum, liver, kidneys and brain was then studied for 24-48 h. The in vitro degradation of Curosurf DPPC by alveolar macrophages was also studied. During the first 3 h after instillation, large amounts of Curosurf left the airways and became associated with tissue, indicating that it mixed rapidly with the endogenous pools of surfactant. A fraction of administered DPPC became associated with the lamellar bodies, suggesting that Curosurf can be recycled. Curosurf administration did not stop the secretion of endogenous surfactant. Very little intact radioactive DPPC could be recovered at any time in alveolar macrophages, however, macrophages have the ability, in vitro, to degrade Curosurf. Newborn rabbits lose Curosurf from the lungs at a slower rate than adult rabbits. One and two days after instillation, organic extracts from the liver, kidney, brain and serum contained small but measurable amounts of radioactivity. These results indicate that Curosurf rapidly enters the pathways of surfactant metabolism and that alveolar macrophages may play an important role in the catabolism of Curosurf. PMID- 9727778 TI - Surfactant composition reflects lung overinflation and arterial oxygenation in patients with acute lung injury. AB - Pulmonary surfactant abnormalities have consistently been documented in patients with acute lung injury (ALI), however, there is little evidence directly correlating them to altered respiratory mechanics. To explore this further, surfactant composition was measured in lung aspirate fluid collected on 15 occasions from 10 patients with ALI. The composition was compared with lung aspirate fluid from 11 intubated patients prior to elective cardiac surgery (CS), and bronchoalveolar lavage fluid from 16 normal subjects. In both the ALI and cardiac groups the proportion of disaturated phospholipids (DSP) and phosphatidylcholine was reduced. Plasma levels of surfactant proteins-A and -B (SP-A and -B) were elevated, but were unrelated to alveolar surfactant levels. In the ALI group, and the ALI + CS group, DSP, normalized to the total phospholipid content, sphingomyelin (SPH), and urea, showed strong direct correlations with arterial oxygen tension/inspiratory oxygen fraction (all p < or = 0.01). In the ALI group, normalized DSP was also directly related to the elastance of the positive end-expiratory pressure-induced increase in the end-expiratory lung volume (all p < or = 0.02), and indirect correlations were found with a measure of lung overinflation (%E2; all p < or = 0.01). We conclude that surfactant composition correlates with lung function abnormalities in acute lung injury and cardiac patients, and that both groups had elevated plasma surfactant proteins-A and -B levels, consistent with a concurrent increase in alveolocapillary permeability. PMID- 9727779 TI - Standardization of ambulatory peak flow monitoring: the importance of recent beta2-agonist inhalation. AB - Standardization of conditions for peak expiratory flow (PEF) monitoring is much more difficult in practice than for laboratory spirometry. Patients are usually asked to record PEF before medication. The aim of this study was to determine the effect of prior bronchodilator use on PEF outcome measures in a clinical trial. Electronic PEF records from 43 subjects with poorly controlled asthma were examined to determine the frequency with which beta2-agonist was inhaled <4 h before PEF measurement, as such PEF are potentially "postbronchodilator". The effect of inclusion of such PEF values on improvement in PEF outcome measures after 8 weeks of inhaled budesonide was calculated. Subjects were asked to record PEF before medication. During run-in, the median frequency of postbronchodilator PEF was 29%, falling to 0% after 8 weeks of budesonide. Inclusion of postbronchodilator PEF led to an overestimation of average morning, evening and daily PEF during run-in (p<0.001). Improvement in these indices with treatment was, therefore, underestimated. Minimum morning PEF expressed as per cent personal best was unaffected. Subjects may not be able to withhold beta2-agonist for 4 h before every peak flow reading. This may change as the level of asthma control changes, leading to a systematic bias in clinical trial end-points or inaccuracy in individual treatment decisions. Simple changes to peak expiratory flow instructions and analysis are proposed. PMID- 9727780 TI - Worldwide variations in the prevalence of asthma symptoms: the International Study of Asthma and Allergies in Childhood (ISAAC) AB - The International Study of Asthma and Allergies in Childhood (ISAAC) was designed to allow comparisons between populations in different countries. ISAAC Phase One, reported here, used standardized simple surveys which were conducted among representative samples of school children from centres in most regions of the world. Two age groups (13-14 and 6-7 yrs) with approximately 3,000 children in each group were studied in each centre. The 13-14 yr olds (n=463,801) were studied in 155 centres (56 countries) and the 6-7 yr olds (n=257,800) were studied in 91 centres (38 countries). There were marked variations in the prevalence of asthma symptoms with up to 15-fold differences between countries. The prevalence of wheeze in the last 12 months ranged from 2.1-32.2% in the older age group and 4.1-32.1% in the younger age group and was particularly high in English speaking countries and Latin America. A video questionnaire completed in the older age group in 99 centres (42 countries) showed a similar pattern. The major differences between populations found in the International Study of Asthma and Allergies in Childhood Phase One are likely to be due to environmental factors. The results provide a framework for studies between populations in contrasting environments which are likely to yield new clues about the aetiology of asthma. PMID- 9727781 TI - Fish consumption and respiratory symptoms among young adults in a Norwegian community. AB - The aim of this study was to investigate the relationship between dietary fish consumption and self-reported respiratory symptoms among young adults. A random sample of 4,300 subjects, aged 20-44 yrs, living in Bergen, Norway, received a postal questionnaire on respiratory symptoms, of whom 80% responded. Mean fish consumption was assessed in a food-frequency questionnaire by asking how often the subject consumed units of fish (150 g) during the last year. Average fish consumption was 1.8 units x week(-1). Fish intake of <1 unit x week(-1) was reported by 24%, 41% reported consumption of 1 unit x week(-1) and 35% intake of >1 unit x week(-1). A high fish intake was significantly associated with increasing age after adjusting for smoking. Adjusted for smoking habits, the prevalence of "cough at night" and "chest tightness" showed a decreasing trend with increasing fish consumption (p<0.05), while such a trend for "wheeze" was demonstrated only in smokers (p=0.008 for interaction). In logistic regression models (adjusting for age, sex, body mass, smoking habits and occupational exposure) fish consumption (three categories) was not significantly associated with "wheeze", "chest tightness", "breathless at night" or "asthma attack", although the odds ratios (OR) were consistently less than 1 (except for "asthma attack"). Fish consumption was of borderline significance as a protective factor of "cough at night", OR = 0.86 (95% confidence interval: 0.76-0.97) but in stratified analyses only in smokers. Subjects reporting very high levels of fish consumption (>14 units x week(-1)) did not have lower prevalences of respiratory symptoms. In conclusion, among young Norwegian adults, with a relatively low prevalence of asthma and an overall high fish intake, fish consumption was not a significant predictor of four out of five respiratory symptoms. PMID- 9727782 TI - Nebulized magnesium sulphate versus nebulized salbutamol in acute bronchial asthma: a clinical trial. AB - Intravenous magnesium sulphate (MgSO4) has successfully been used in the treatment of acute asthma. The present study investigated the efficacy of nebulized MgSO4 as a bronchodilator in acute asthma as compared to nebulized salbutamol. This was a randomized, double-blind, controlled clinical trial. Asthmatics aged 12-60 yrs in acute exacerbation, with a peak expiratory flow (PEF) <300 L x min(-1), not having taken bronchodilators and not requiring assisted ventilation were included. Patients were randomized to receive treatment with serial nebulizations of either 3 mL (3.2% solution, 95 mg) MgSO4 solution or 3 mL (2.5 mg) salbutamol solution. All patients were also given 100 mg hydrocortisone i.v., and were monitored continuously for 2 h after which they were given supplemental treatment (if and when needed) and either discharged or admitted. Fischl index, PEF improvements (in % predicted) and admission rates were the outcome variables. Thirty-three patients were studied. Fischl score improvement was comparable and significant in both groups (4.31 to 0.43 in the MgSO4 group and 4.29 to 0.76 in the salbutamol group). The increase in PEF was statistically significant and comparable in both groups (by 35% pred in the MgSO4 and by 42% pred in the salbutamol group). Two patients warranted admission in the salbutamol group and one in the MgSO4 group. Nebulized MgSO4 had a significant bronchodilatory effect in acute asthma. This effect was not significantly different from that of nebulized salbutamol. PMID- 9727783 TI - Evidence for mast cell activation during exercise-induced bronchoconstriction. AB - Controversy remains about the causative mediators in the bronchoconstrictive response to exercise in asthma. This study examined whether mast cell activation is a feature of exercise-induced bronchoconstriction by measuring urinary metabolites of mast cell mediators. Twelve nonsmoking subjects with mild asthma and a history of exercise-induced bronchoconstriction exercised on a stationary bicycle ergometer for 5 min at 80% maximum work load. Pulmonary function was monitored and urine was collected before and 30 and 90 min after the provocation. The urinary concentrations of the mast cell markers 9alpha,11beta-prostaglandin (PG)F2 and Ntau-methylhistamine, as well as leukotriene E4 (LTE4) were determined by immunoassay. Seven of the 12 subjects (responders) experienced bronchoconstriction (>15% fall in the forced expiratory volume in one second) following exercise, whereas the pulmonary function of the remaining five subjects (nonresponders) remained stable. The urinary excretion (mean+/-SE) of 9alpha,11beta-PGF2 in the responders increased significantly compared with the nonresponders at 30 (77.1+/-14.4 versus 37.2+/-5.6; p<0.05) and 90 min (79.3+/ 8.6 versus 40.4+/-8.5, p<0.05) after exercise challenge. The urinary excretion of Ntau-methylhistamine and LTE4 was not significantly different between the two groups at 30 or 90 min after exercise. The findings represent the first documentation of increased urinary levels of 9alpha,11beta-prostaglandin F2 in adults following exercise challenge and provides clear evidence for mast cell activation during exercise-induced bronchoconstriction in asthmatics. PMID- 9727784 TI - Influence of antimicrobial chemotherapy on spirometric parameters and pro inflammatory indices in severe pulmonary tuberculosis. AB - Patients who have completed a treatment for severe pulmonary tuberculosis (TB) are often left with severe respiratory disability. There have been few prospective studies assessing the effect of treatment on lung function in such patients. The influence of antimicrobial chemotherapy on lung function was investigated over a six month period in patients with newly diagnosed pulmonary TB to test the hypothesis that treatment improves lung function, as well as to identify factors that may influence lung function outcome. Seventy-six patients were recruited into the study, of whom 74 completed the treatment programme. Forty-two were current smokers and 13 seropositive for the human immunodeficiency virus. Improvement in lung function occurred in 54% of patients, but residual airflow limitation or a restrictive pattern was evident in 28% and 24% of patients, respectively. The extent of lung infiltration (radiographic score) both at the outset and after chemotherapy was significantly and negatively related to forced expiratory volume in one second (FEV1) (% pred) (r=-0.41, and r=-0.46, respectively). The post-treatment serum C-reactive protein and alpha1-protease inhibitor levels were negatively associated with FEV1 (% pred) (r=-0.30 and r= 0.35, respectively). These findings demonstrate that, while antimicrobial chemotherapy may lead to improved lung function in patients with pulmonary tuberculosis, a large proportion of patients has residual impairment. The most significant factor influencing post-treatment lung function status, as measured by forced expiratory volume in one second (% predicted), is the pretreatment and post-treatment radiographic score, which acts as a marker of the extent of pulmonary parenchymal involvement in tuberculosis. PMID- 9727785 TI - Thoracic infection caused by Streptococcus milleri. AB - The objective of this study was to increase our understanding of the importance of members of the Streptococcus milleri (SM) group as respiratory pathogens, by studying the epidemiological and clinical features of thoracic infections caused by this group and comparing the epidemiology and prognosis of empyema caused by SM with cases of pneumococcal aetiology. The clinical histories and microbiology reports were reviewed in 27 cases of thoracic infection caused by SM over a period of 8 yrs. Cases of pneumococcal empyema that occurred during the same period were also analysed. Diagnoses were made of cases of empyema, including six with pneumonia and one with pulmonary abscess, three cases of pneumonia and two of mediastinitis. In 17 cases, SM was the only pathogen isolated. There was a history of instrument or surgical procedures on the digestive or respiratory tract in 59%. Secondary bacteraemia was documented in three cases. The treatment administered, a combination of antibiotics and surgery, was successful in 22 of 27 (81%) of cases. All strains were susceptible to penicillin. When the characteristics of the empyemas caused by monomicrobial SM infection were compared with those of pneumococcal aetiology from the same period of study, significant differences were found with respect to age, origin of the infection and the need for surgery. In conclusion, thoracic infections caused by Streptococcus milleri are largely pleural. They are polymicrobial in one-third of cases, commonly acquired in hospital and, in most patients, associated with major surgery and/or surgical procedures of the respiratory or digestive tract. The empyema frequently requires thoracotomy for complete resolution. PMID- 9727786 TI - Randomized controlled trial of pulmonary rehabilitation in severe chronic obstructive pulmonary disease patients, stratified with the MRC dyspnoea scale. AB - This study tested the hypothesis that severity of respiratory disability may affect the outcome of pulmonary rehabilitation. In this randomized, controlled study, 126 patients with chronic obstructive pulmonary disease (COPD) were stratified for dyspnoea using the Medical Research Council (MRC) dyspnoea score into MRC3/4 (Moderate) (n=66) and MRC 5 (Severe) dyspnoeic (n=60) groups. The patients were randomly assigned to an eight week programme of either exercise plus education (Exercise group) or education (Control group). Education and exercise programmes for the moderately dyspnoeic patients were carried out in a hospital outpatient setting. Severely dyspnoeic patients were all treated at home. Those in the Exercise group received an individualized training programme. There was a significant improvement in shuttle walking distance in the moderate dyspnoeic group, who received exercise training; baseline (mean+/-SEM) 191+/-22 m, post-rehabilitation 279+/-22 m (p<0.001). There was no improvement in exercise performance in the severely dyspnoeic patients receiving exercise. Neither group of control patients improved. Health status, assessed by the Total Chronic Respiratory Disease Questionnaire score, increased in the moderately dyspnoeic patients receiving exercise from 80+/-18 to 95+/-17 (p<0.0001) after rehabilitation. Much smaller changes were seen in the other three groups. Improvement in exercise performance and health status in patients with chronic obstructive pulmonary disease after an exercise programme depends on the initial degree of dyspnoea. PMID- 9727787 TI - Emotional status does not alter exercise tolerance in patients with chronic obstructive pulmonary disease. AB - Exercise tolerance in chronic obstructive pulmonary disease (COPD) patients has been shown to be related to airway limitation and dyspnoea, but little is known about the effects of an emotional status on physical performance. We examined 49 COPD patients with a wide spectrum of airway limitation severity and hypoxaemia. Exercise tolerance was evaluated using the Six-minute Walking Distance Test (6MWD), dyspnoea at rest and on exercise was measured using the visual analogue scale, and the emotional status was evaluated using the battery of psychological tests. The average 6MWD (mean+/-SD) was 355+/-112 m. In the majority of patients a fall in arterial blood oxygen saturation (Sa,O2) on exercise of >3% was found. The mean dyspnoea score of 21+/-19 at rest increased to 66+/-19 on exercise. All subjects demonstrated an increased level of anxiety. The majority also demonstrated elevated emotional tension, and half of the study group showed signs of depression. Step-wise multiple regression analysis with results of 6MWD as dependent and other studied variables as independent variables showed that exercise tolerance depended mainly on airway limitation. The forced expiratory volume in one second (FEV1) explained 24% of the variance. The forced vital capacity added a further 10%, and arterial blood carbon dioxide tension contributed 7%. The dyspnoea level on exercise added only 0.9%. All four variables explained 42% of the variance. There was no correlation between 6MWD and any of the variables characterizing the emotional status. We conclude that the emotional status of chronic obstructive pulmonary disease patients is characterized by an increased level of psychological tension, anxiety and depression, but these do not affect exercise tolerance as assessed by the six minute walking distance test. PMID- 9727788 TI - The haemodynamic response to exercise in chronic obstructive pulmonary disease: assessment by impedance cardiography. AB - This study aimed to determine the differences in haemodynamic responses to a standard incremental exercise test between outpatients with chronic obstructive pulmonary disease (COPD) and age-matched controls and to discover the relationship between severity of airflow obstruction and exercise haemodynamics in COPD. Twenty-two male patients with COPD (forced expiratory volume in one second (FEV1)/vital capacity (VC))<80% predicted) and 20 age-matched male controls performed an incremental exercise test (10 W x min(-1)) with ventilatory function and changes in stroke volume (deltaSV) and cardiac output (deltaCO) measured by means of electrical impedance cardiography (EIC). Submaximal deltaSV and deltaCO were lower in COPD patients. Peak exercise deltaSV were equal in patients and controls (128+/-33 versus 129+/-29%, p=0.98), whereas peak deltaCO was lower in patients (COPD versus controls: 232+/-71 versus 289+/-54%, p<0.005). In COPD patients, FEV1 (% pred) was significantly correlated to deltaSV at all submaximal exercise intensities, to peak exercise deltaSV and to peak exercise deltaCO. FEV1/VC (% pred) was significantly correlated to deltaSV at 30 and 60 W. In conclusion, in chronic obstructive pulmonary disease an aberrant haemodynamic response to exercise was found, especially in patients with severe airflow obstruction. This aberrant response is related to the degree of airflow obstruction and may limit exercise performance in patients with severe chronic obstructive pulmonary disease. PMID- 9727789 TI - Inflammatory cells and mediators in bronchial lavage of patients with chronic obstructive pulmonary disease. AB - Cigarette smoking is the most important cause of chronic obstructive pulmonary disease (COPD). Although the precise sequence of events that leads a smoker to experience airway obstruction is not completely clear, airway inflammation is a relevant factor. To investigate airway inflammation, 12 nonatopic smoking COPD patients with a forced expiratory volume in one second (FEV1) < or = 75% predicted and 10 normal nonsmoking subjects (NS) were studied with bronchoscopy and bronchial lavage (BL). Serum immunoglobulin (Ig)E levels of COPD patients correlated with the smoking history (r=0.7, p=0.008). In BL of COPD patients there was an increase of neutrophils (median, range) (COPD 62.6x10(3), 1.2-323, NS 1.35, 0-19.2, p=0.001), eosinophils (COPD 1.6, 0-6.9, NS 0.15, 0-3.7, p=0.035), the levels of interleukin (IL)-8 (COPD 1079 pg x mL(-1), 121-2,500, NS 20.4, 7.2-59, p=0.001), myeloperoxidase (MPO) (COPD 752 microg x L(-1), 11-5,500, NS 22.1, 8-70, p=0.001) and eosinophil cationic protein (ECP) (COPD 21.5 microg x L(-1), 1.8-161, NS 2, 1.8-4.9, p=0.001). Significant correlations were found in BL of COPD patients between IL-8 and neutrophils (p=0.02), MPO and neutrophils (p=0.02), IL-8 and MPO (p=0.0001) and ECP and eosinophils (p=0.02). In addition, the ratios between the BL levels of MPO and the number of neutrophils and between ECP levels and eosinophils were higher in COPD patients than in NS (p=0.03 and 0.01, respectively). These data suggest that cigarette smoke is associated with increased amounts of airway interleukin-8, a chemotactic factor for neutrophils and eosinophils. Recruited neutrophils and eosinophils are activated and they release increased amounts of inflammatory mediators capable of damaging the bronchial tissue. PMID- 9727791 TI - Cigarette smoke inhalation and lung damage in smoking volunteers. AB - Cigarette smoking is the dominant risk factor for chronic obstructive pulmonary disease (COPD) but only 10-15% of smokers develop the condition. Risk does not relate closely to cumulative cigarette consumption, perhaps because smokers vary in the degree and depth of smoke inhalation. This study examined the role of smoke inhalation in the development of COPD. Eighty current smokers and 20 lifetime nonsmoking volunteers (aged 35-65 yrs) were recruited. Lung function variables were measured and high-resolution computed tomography (HRCT) scans performed. Smoke inhalation was assessed by CO boost (the increment of expired carbon monoxide 5 min after smoking a cigarette) and serum cotinine. Mean CO boost was 6.3 parts per million (ppm) in smokers with low CO transfer coefficients (KCO) and 2.9 ppm in those with normal KCO (p=0.006); 7.2 ppm in smokers with both HRCT-defined emphysema and a low KCO and 2.6 ppm in those with neither abnormality (p=0.002); 4.5 ppm in smokers with HRCT-defined emphysema alone and 2.8 ppm in those without (p=0.08). Mean serum cotinine was 328 ng x mL( 1) in smokers with chronic productive cough and 243 ng x mL(-1) in those without (p=0.005). Lifetime nonsmokers had normal HRCT scans, lung function and serum cotinine. Emphysema is associated with high alveolar smoke exposure as measured by CO boost. Productive coughing is associated with high nicotine uptake, probably from airway smoke particle deposition. PMID- 9727790 TI - Eosinophilic airway inflammation induced by repeated exposure to cigarette smoke. AB - Acute exposure to cigarette smoke causes airway hyperresponsiveness (AHR) in guinea-pigs, which resolves within a few hours. Repeated exposure may have a different effect on the airways. To address this question, guinea-pigs were repeatedly exposed to cigarette smoke (six cigarettes for 1 h x day(-1)) for 14 consecutive days. Airway responsiveness to inhaled histamine and differential cell counts in bronchoalveolar lavage fluid (BALF) were evaluated 1 day after the last exposure. Significant neutrophilia in BALF was observed after 3 days of smoke exposure. Significant eosinophilia in BALF and AHR were observed after 14 days of smoke exposure, but not after 3 or 7 days of smoke exposure. These changes persisted until 3 days after the last exposure and resolved 7 days afterwards. Histologically, the recruited eosinophils were observed predominantly in the airways, but not in the alveoli. Treatment with E-6123, a specific platelet-activating factor receptor antagonist (1 mg x kg(-1) x day(-1) p.o. during smoke exposure) significantly inhibited the eosinophil influx and AHR. Repeated exposure to cigarette smoke may induce prolonged airway inflammation and airway hyperresponsiveness in guinea-pigs. Platelet-activating factor or platelet activating factor-like lipids may play a key role in airway hyperresponsiveness, presumably by the induction of eosinophilic airway inflammation. PMID- 9727792 TI - Regulation of breathing in hyperthyroidism: relationship to hormonal and metabolic changes. AB - We sought to examine the breathing pattern, inspiratory drive and chemosensitivity of hyperthyroid patients and to explore the interactions between their thyroid hormones, basal metabolism and chemosensitivity. We studied 15 hyperthyroid patients and 15 sex- and age-matched controls. Thyroid hormone levels, arterial blood gas tensions, lung volumes, diffusing capacity for CO, maximal respiratory pressures and oxygen uptake measurements were performed. Breathing pattern and mouth occlusion pressure (P0.1), as well as ventilatory and P0.1 responses to hyperoxic progressive hypercapnia and isocapnic progressive hypoxia, were also evaluated. Compared with the control subjects, the hyperthyroid patients showed significantly lower resting arterial CO2 tension, tidal volume and significantly higher mean inspiratory flow and P0.1. Ventilatory and P0.1 responses to CO2 and hypoxia were also greater in the hyperthyroid patients than in the control group. All these changes returned to normal after treatment. In the patients, significant relationships between tri-iodothyronine and P0.1, P0.1 response to hypoxia, and P0.1 response to hypercapnia were found. In contrast, in hyperthyroidism there was no relationship between oxygen uptake and P0.1 response to hypoxia. We conclude that hyperthyroid patients exhibit a significant relationship between their thyroid hormone levels and their increased inspiratory drive and chemosensitivity. PMID- 9727793 TI - Effects of high-altitude periodic breathing on sleep and arterial oxyhaemoglobin saturation. AB - This study aimed to investigate the effect of periodic breathing (PB) at high altitude on sleep structure and arterial oxygen saturation (Sa,O2). Five healthy subjects underwent polysomnographic studies at sea level, and during the first and the fourth week of sojourn at 5,050 m. Their breathing pattern, sleep architecture and Sa,O2 were analysed. PB was detected in the high-altitude studies during nonrapid eye movement (NREM) sleep and tended to increase from the first to the fourth week. Stages 3-4 were absent in four subjects at the first week, but only in one at the fourth week, irrespective of the amount of PB. The arousal index was 11.6+/-3.8 at sea level, 30.1+/-15.5 at the first week at altitude and 33.0+/-18.2 at the fourth week. At altitude, arousal index in NREM sleep was higher during PB than during regular breathing. In NREM sleep, the mean highest Sa,O2 levels in NREM epochs with PB were higher than in those with regular breathing by 2.8+/-1.7% at the first week and 2.9+/-1.5% at the fourth week (p<0.025). From the first to the fourth week, mean Sa,O2 increased significantly during wakefulness (5.6%), NREM (5.2% with regular breathing and 5.3% with PB) and rapid eye movement sleep (7.6%). The data demonstrate a slight role of periodic breathing in altering sleep architecture at high altitude and also show that periodic breathing induces only a minor improvement in arterial oxygen saturation during nonrapid eye movement sleep. PMID- 9727794 TI - Treatment of Cheyne-Stokes respiration with nasal oxygen and carbon dioxide. AB - Cheyne-Stokes respiration (CSR) is common in patients with congestive heart failure (CHF) and is associated with significant nocturnal O2 desaturation, arousals and sympathetic activation. Nocturnal O2 reduces CSR by only about 50%. More complete suppression of CSR may be achieved by adding CO2 to O2. This study therefore aimed to evaluate the effects of nocturnal O2 plus CO2 on CSR, sleep and sympathetic activation. Nine patients with CHF (age 59+/-5 yrs; left ventricular ejection fraction 17.8+/-1.2% (mean+/-SEM) were studied in a cross over, single-blind, placebo-controlled trial. After an accommodation night the patients were randomly assigned to one night each of O2 plus CO2 as well as air applied by nasal prongs. Nocturnal O2 plus CO2 reduced the duration of CSR as percentage of total sleep time (48.0+/-10 versus 7.4+/-2.0%; p=0.008) and increased arterial oxygen saturation (Sa,O2) as well as mean transcutaneous carbon dioxide tension (Ptc,CO2) (5.2+/-0.3 kPa (39+/-2 mmHg) versus 5.7+/-0.3 kPa (43+/-2 mmHg) p=0.011). Sleep did not improve and arousals were not reduced. Plasma noradrenaline was higher on the treatment night (486+/-116 versus 669+/ 163 ng x L(-1); p=0.035). In conclusion, nocturnal O2 plus CO2 improves Cheyne Stokes respiration in patients with congestive heart failure but has adverse effects on the sequel of Cheyne-Stokes respiration, namely sympathetic activation. PMID- 9727795 TI - Effect of oxygen on breathing irregularities during haemodialysis in patients with chronic uraemia. AB - Hypoxaemia and breathing irregularities have been shown to occur during haemodialysis in patients with chronic renal failure. This study examined the role of hypoxia in the genesis of the irregular breathing during haemodialysis. The ventilatory patterns using respiratory inductance plethysmography and arterial blood gases were studied in seven males with chronic renal failure on long-term haemodialysis. The study was carried out before and during dialysis on one day without (D1) and another day with intranasal oxygen at 4 L x min(-1) (D2). On D1, mean (SD) arterial oxygen tension (Pa,O2) fell 1.9 (0.9) kPa (p<0.001) and mean minute ventilation (V'E) fell 1.9 (1.1) L x min(-1) (p<0.01) during dialysis. The arterial carbon dioxide tension (Pa,CO2) did not show a significant decrease (4.7 (0.2) kPa before and 4.6 (0.2) kPa during dialysis). Cumulative number of apnoeas was 64 and the coefficients of variation (COV) of respiratory frequency (fR) and tidal volume (VT) were 29.6 (11.9) and 38.2 (11.9)%, respectively. On D2, mean Pa,O2 remained stable (20.4 (4.1) kPa before, 21.3 (4.1) kPa during dialysis). There was no significant change in mean V'E (6.4 (0.9) L x min(-1) before, 5.5 (0.5) L x min(-1) during dialysis). Pa,CO2 decrease was not significant but the fall was greater (4.8 (0.1) kPa before, 45 (0.5) kPa during dialysis). Cumulative number of apnoeas was 94 and the COVs offR and VT were 35.8 (5.1) and 40.5 (11.3)%, respectively. Oxygen administration did not significantly affect the haemodialysis-induced changes in ventilation and breathing pattern, despite a significant protective effect from the fall in arterial oxygen tension. It was concluded that the fall in arterial oxygen tension is not the main determinant of breathing irregularities during haemodialysis. PMID- 9727796 TI - Cough and sleep in inner-city children. AB - This study aimed to determine whether cough at night keeps children awake, to describe the relationship between children's cough and sleep and to report parents' perceptions of their children's cough and sleep. Thirty-nine children with reported persistent cough at night (>3 weeks) were recruited and studied for 6 nights by video-recording. Coughs were counted and sleep state was coded for awake, restless sleep and quiet sleep. The relationships between cough and sleep state between subjects and within subjects were examined by correlation and regression. After night 2 the parents were asked whether their child had coughed or had disturbed sleep and after night 6 they were asked whether there had been any change. There was a weak relationship between log percentage of the night awake and log number of coughs (r=0.13, SE 0.036), and log (percentage of the night awake plus restless sleep) and log number of coughs (r=0.016, SE 0.0071). If the relationship between cough and sleep state is causal, halving the number of coughs will reduce the percentage of the night awake by 9% (95% confidence interval (CI) 4-15%) and percentage awake and restless by 1% (95% CI 0-2%). All but one parent correctly identified coughing and 82% detected change. Most could not comment on their child's sleep. Improvement in cough would result in little reduction in either the percentage of the night awake or awake and restless in the average child in the population studied. Parents could detect whether their children were coughing but not whether their sleep was disturbed. PMID- 9727797 TI - Prevalence of childhood asthma, rhinitis and eczema in Scandinavia and Eastern Europe. AB - There is evidence that the prevalence of allergies and asthma differs between populations in western and eastern Europe. This study investigated the prevalence of wheezing, rhinitis and eczema among schoolchildren in urban and rural areas of Scandinavia and the formerly socialist countries of Eastern Europe. A total of 79,000 children from two age groups (13-14 yrs and 6-7 yrs) in 18 study centres responded to a questionnaire within the International Study of Asthma and Allergy in Children (ISAAC). The 12 month period prevalence of symptoms of asthma, allergic rhinoconjunctivitis and atopic eczema was calculated. The prevalence of wheezing among the 13-14 yr old children was 11.2-19.7% in Finland and Sweden, 7.6-8.5% in Estonia, Latvia and Poland and 2.6-5.9% in Albania, Romania, Russia, Georgia and Uzbekistan (except Samarkand). The prevalence of itching eyes and flexural dermatitis varied in a similar manner between the three regions. The regional differences were less pronounced among the 6-7 yr old children in the seven participating centres. The highest prevalence of rhinitis was recorded in April-July in Scandinavia and during the winter months in the other countries. The prevalence of atopy-related disorders was higher in Scandinavia than in Estonia, Latvia and Poland, which in turn had a higher prevalence than five other countries of eastern Europe with a culture less similar to western Europe. This supports the hypothesis that "Western life style" is associated with a high prevalence of childhood allergy. PMID- 9727798 TI - Bronchodilator response in 3-6.5 years old healthy and stable asthmatic children. AB - Few data are available on the bronchodilator response in preschool children. This study was set up to study baseline lung function and bronchodilator responses in healthy and asthmatic children younger than 7 yrs old. In 281 preschool children attending kindergarten (age range 2.7-6.6 yrs old) respiratory system resistance (Rrs) and reactance (Xrs) by impulse oscillation system at 5, 10, 15, 20, 25 and 35 Hz as well as resonance frequency (f0) were measured before and 20 min after 200 microg inhaled salbutamol by a metered-dose inhaler connected to a spacer device. Thirty-four of them were diagnosed as asthmatics based on a validated standardized questionnaire. The mean Rrs (+/-SD) at 5 Hz (Rrs,5) was 1.03 (+/ 0.24) kPa x L(-1) x s for healthy children and 1.09 (+/-0.26) kPa x L(-1) x s for stable asthmatics. The mean change in Rrs,5 after salbutamol was -0.13 (+/-0.20) kPa x L(-1) x s for the healthy children and -0.09 (+/-0.25) kPa x L(-1) x s for the asthmatic group. The scatter for the measurements was striking. Neither baseline values of impulse oscillation nor its changes after bronchodilator administration was significantly different between healthy and stable asthmatic children. A change in respiratory system resistance at 5 Hz of 40% is to be considered the cut-off for a " positive" bronchodilator response. PMID- 9727799 TI - Diffuse panbronchiolitis in rheumatoid arthritis. AB - The association of progressive obliterative bronchiolitis (OB) with rheumatoid arthritis (RA) is uncommon but has been reported previously. Diffuse panbronchiolitis (DPB) is a unique inflammation principally affecting the respiratory bronchioli and has been reported mainly in Japanese adults. Recently, DPB has also been noted in patients with RA in Japan. Therefore, there might be considerable overlap in clinical features between DPB and OB associated with RA in Japan. The aim of this study was to evaluate the clinicopathological characteristics of bronchiolitis in patients with RA. Three RA patients clinically diagnosed as having DPB were evaluated. All patients underwent chest radiographs, pulmonary function tests (PFT) and post mortem examination. Clinical features in all patients were a history of productive cough, exertional dyspnoea, wheezing and/or coarse crackles. Chest radiographs showed small nodular shadows up to 2 mm in diameter with bronchiolectasis throughout both lungs in all patients. The PFT revealed marked obstructive impairment in all patients. All patients died of progressive respiratory failure. Pathologically, two out of the three cases were confirmed as DPB, while the remaining one case was confirmed as OB, because the primary obstructive lesions were in the respiratory bronchioli in the former and in the membranous bronchioli and the proximal small bronchi in the latter. Thus, the clinical features of DPB and OB were strikingly similar, but the histopathological features revealed distinct differences. This study demonstrated that there was considerable overlap in clinical features between diffuse panbronchiolitis and obliterative bronchiolitis associated with rheumatoid arthritis, suggesting that diffuse panbronchiolitis might be a new manifestation of rheumatoid arthritis. The differentiation of these two disease entities is significant in making decisions on their therapeutic modality and is possible by analysing the precise histopathological findings of the lung. PMID- 9727800 TI - Chest radiographic staging in allergic bronchopulmonary aspergillosis: relationship with immunological findings. AB - The question of whether a chest radiographic severity staging system could be correlated with standard blood/serum diagnostic indices in allergic bronchopulmonary aspergillosis (ABPA) was addressed in 41 patients. Asthma and positive Aspergillus fumigatus (AF) serology were considered essential diagnostic inclusion criteria. Eosinophil count, serum immunoglobulin (Ig)E and immediate skin hypersensitivity were also tested to grade patients as "definite" or "likely" ABPA. Definite cases had all five of these factors present, whereas likely cases had three or more. Chest radiographs were examined by experienced radiologists blinded to the clinical data. The six-stage radiographic score (0-5) was based on the severity and duration of changes seen: stage 0: normal; stage 1: transient hyperinflation; stage 2: transient minor changes; stage 3: transient major changes; stage 4: permanent minor changes; and stage 5: permanent major changes. Significant positive correlations (p<0.05) were observed between peak AF titres (expressed as an index), peak eosinophil count and radiographic severity stage. When considered as subgroups, these correlations approached, but did not reach, significance for the group with "likely" ABPA (n=28), but in the group with definite ABPA (n=13), there was a high correlation between radiographic score and peak AF index (r=0.59), as well as peak eosinophil count (r=0.62). This study suggests that the peak Aspergillus fumigatus index and eosinophil counts correlate best with the severity of radiographic stages in allergic bronchopulmonary aspergillosis. This chest radiographic staging system may be useful in the clinical assessment and management of patients with allergic bronchopulmonary aspergillosis, particularly in those patients with more severe radiographic stages. PMID- 9727801 TI - Assessment of accuracy and applicability of a new electronic peak flow meter and asthma monitor. AB - The aim of this study was to assess the accuracy and applicability of a portable electronic peak flow meter combined with an asthma monitor (AM1, Jaeger, Germany) which measures peak expiratory flow (PEF), forced expiratory volume in one second (FEV1) and forced vital capacity (FVC). The technical accuracy in PEF, FEV1 and FVC measurement was tested according to American Thoracic Society (ATS) criteria for monitoring devices using a flow generator. In addition, the effect of connecting a heated screen pneumotachograph (PT) to the AM1 was determined and the accuracy in FEV1 determinations was evaluated by simultaneous measurements in 49 normal volunteers. The devices tested fulfilled all ATS criteria for monitoring devices with respect to the accuracy of PEF, FEV1, and FVC measurements. The conditions of intra- and interdevice variability were satisfied in all cases. Compared with the PT, the AM1 showed about 4% lower values in FEV1, as measured in the 49 subjects. In conclusion, the electronic peak flow meter and asthma monitor AM1 yielded valid measurements of peak expiratory flow and forced expiratory volume in one second, which matched the accuracy criteria of the American Thoracic Society standards for monitoring devices. PMID- 9727802 TI - Drug output from nebulizers is dependent on the method of measurement. AB - The objective of this study was to determine whether current regulatory methods for assessing the output of nebulizers are appropriate for the delivery of nebulized steroid suspensions to patients. We studied a conventional jet nebulizer (the Intersurgical Cirrus), an open-vent nebulizer (the Medicaid Sidestream) and a breath-enhanced nebulizer (the Pari LC Plus), using a constant sampling flow or a sinusoidal pump to represent the breathing pattern of children from 6 months to adulthood. Recovery of budesonide released from the nebulizers onto filters was reduced when using breathing simulation compared with constant flow, and this reduction was greatest for the conventional nebulizer (Cirrus, 103 microg with constant flow to 4.4 microg with a 50 mL tidal volume; Pari, 176 microg to 25 microg). The open-vent nebulizer deposited very little budesonide on the filter at lower tidal volumes (4.5 microg with a 50 mL tidal volume), possibly because the enhanced flow of aerosol laden air was greater than the inspiratory flow from the breathing simulator. The output of the LC plus was reduced at high flow, from 176 microg at 20 L x min(-1) to 93 microg at 60 L x min(-1). Overall, the measured output varied by up to 700%, depending on the method used. These results suggest that breathing patterns dramatically alter the measured output of different nebulizers and that breathing simulation should be included as part of their assessment. PMID- 9727803 TI - A new method for measuring aerosol nebulizer output using radioactive tracers. AB - Reproducibility and comparability of bronchial challenge tests depends critically on accurate assessment of nebulizer output. Evaporation during nebulization means that simple weighing is inaccurate, overestimating the delivered dose of active ingredient. We wanted to quantify this effect in the context of intermittent nebulization, using a dosimeter as used in bronchial provocation tests. Output of three types of nebulizer, from the MEFAR dosimeter, was measured by radioactive tracer, using a standard solution of technetium-99m-pertechnetate (1.5 kBq x mL( 1)) in 4 mL of normal saline. The aerosol was impacted by suction onto a microfilter, and the radioactivity measured. Nebulizers were weighed before and after nebulization. Ratio of nebulized volume calculated from the radioactivity on the filter, to the total volume loss by weight, was expressed as nebulized ratio. The effect on output of two concentrations of methacholine, two tracers of different weights, and change in temperature, were assessed. Nebulized ratio varied between 44.1-71.6%. Results were more consistent within the same type of nebulizer than between different makes. Neither changes in molar concentration nor molecular weight affected nebulizer output or nebulized ratio. Mean nebulized ratio was 58.5%, showing that calibration by weighing, overestimates the delivered dose by a factor of approximately two. Measuring radioactivity eluted from a microfilter, onto which nebulized output had been impacted proved to be a satisfactory method of calibration. PMID- 9727805 TI - Development and validation of a screening test for 12 common mutations of the cystic fibrosis CFTR gene. AB - The results obtained using deoxyribonucleic acid (DNA) amplification-based tests must be accurate and reproducible. One such test that simultaneously detects any of 12 of the most common mutations of the cystic fibrosis transmembrane conductance regulator gene is presented in this report. An investigation was conducted into how changes of primer, DNA template and Taq DNA polymerase concentrations and of polymerase chain reaction annealing temperatures affect the test. A total of 383 DNA samples obtained from different laboratories was then examined. The preliminary studies defined the conditions under which accurate results are obtained even if the test is performed under suboptimal conditions. Subsequently, 377 (98.4%) of the DNA samples analysed were in full agreement with DNA typing results derived by other methods. The remaining 1.6% of samples were not mistyped, rather they were not scored owing to failure to detect control DNA sequences. These were also archival DNA preparations rather than freshly prepared samples from venous blood. Careful primer design and optimization of reaction conditions are important in the development of multiplex deoxyribonucleic acid amplification-based diagnostic tests. Providing the recommended protocols are followed, the test described here is simple to carry out, gives accurate results and works well if performed within defined operational windows for each reaction variable. PMID- 9727804 TI - In vitro performance of three combinations of spacers and pressurized metered dose inhalers for treatment in children. AB - The performance of pressurized metered dose inhalers (pMDIs) and spacers in correct dose recommendations is important, but limited information on dose delivery and fine-particle dose from different combinations of spacers and pMDIs is available. In this study, three combinations of spacers and pMDIs were investigated: NebuChamber and AeroChamber with budesonide pMDI and Babyhaler with fluticasone propionate pMDI. Doses were withdrawn onto a filter either with a breathing simulator (dose to ventilator) or with constant flow (maximal dose). The fine-particle dose was assessed with a cascade impactor (Andersen Sampler). The effect of repeated use and cleaning of the spacers on the passive fallout of aerosol within the spacers was determined by evacuating the dose on a filter 2, 5, 10 and 30 s after actuating the spray. The drugs were quantified by liquid chromatography. The NebuChamber delivered the highest doses, both maximal dose and dose to ventilator. The recovered doses (means+/-SD) were 55+/-6% and 51+/ 2%, respectively, of the delivered dose from the pMDI. The corresponding results for the Babyhaler were 41+/-7% and 24+/-4% and for the Aerochamber 27+/-3% and 17+/-3%. The passive fallout of aerosol, determined as half-life (t1/2) was around approximately 30 s for the NebuChamber, 9-15 s for the Babyhaler and approximately 10 s for the AeroChamber. The present study confirms that there are significant differences in dose output from different combinations of pressurized metered dose inhalers and spacers, with the NebuChamber giving the highest dose, both as delivered dose and in droplets <4.7 microm. Interactions with the spacer material, dead space in the inspiratory line and entrainment of air during inhalation due to inefficient valve control could account for these differences. PMID- 9727806 TI - Hydrogen peroxide in exhaled air of healthy children: reference values. AB - An increased content of hydrogen peroxide (H2O2), a marker of inflammation, has been described in the condensate of exhaled air from adults and children with inflammatory lung disorders, including asthma. However, the normal range of [H2O2] in the exhaled air condensate from healthy children has not been established. Therefore, the aim of this study was to determine the reference range of exhaled [H2O2] in healthy school-aged children. Ninety-three healthy nonsmoking children (48 female and 45 male, mean age 10 yrs, range 8-13 yrs), with a negative history for allergy, eczema or respiratory disease and with a normal lung function, participated. Exhaled air condensate was examined fluorimetrically for the presence of H2O2. In addition, the reproducibility of [H2O2] within subjects and between days and the stability of [H2O2] during storage at -20 degrees C were assessed. The median [H2O2] in the exhaled air condensate of all children was 0.13 microM, with a 2.5-97.5% reference range of <0.01-0.48 microM. No significant difference existed between males and females. There was no correlation between exhaled [H2O2] and age or lung function. Repeated [H2O2] measurements on 2 consecutive days showed satisfactory within subject reproducibility and [H2O2] in stored samples remained stable for at least 1 month at -20 degrees C. In conclusion, this study provides reference data for exhaled hydrogen peroxide in a large group of healthy children. The observed levels were lower than those reported previously for healthy adults and were independent of age, sex and lung function. PMID- 9727807 TI - Quality-of-life assessment in children and adolescents with asthma. AB - Health-related quality of life has become an essential part of health outcome measurement in chronic disorders. However, it is only recently that health professionals have focused on quality-of-life assessment in children and adolescents. Several generic, as well as the asthma-specific quality-of-life instruments specifically designed for use in children and adolescents are reviewed in this article with particular regard to the conceptual and methodological features of the measures and their applicability in clinical studies. The recently published Child Health Questionnaire is a useful generic instrument to comprehensively assess quality of life, in particular when comparing young people with different chronic disorders. The Pediatric Asthma Quality-of-life Questionnaire has shown responsiveness to change over time, but it lacks age-specificity with regard to psychosocial issues and comprehensiveness of quality-of-life assessment. In contrast, the Childhood Asthma Questionnaire provides three different versions for different target ages. However, its generic part is not reflective of the respondent's health status. The other asthma specific instruments have major conceptual deficiencies when used as a single measure for quality-of-life assessment. In the absence of a single ideal instrument, the use of batteries of quality-of-life instruments is therefore recommended and further research is required to identify the impact that age and developmental status have on quality-of-life assessment. PMID- 9727808 TI - Central sleep apnoea in congestive heart failure despite vagal denervation after bilateral lung transplantation. AB - Nonhypercapnic central sleep apnoea is a disorder of respiratory control characterized by hyperventilation previously attributed to the stimulation of either pulmonary vagal afferent nerve fibres or respiratory chemoreceptors. This report describes central sleep apnoea in a patient with congestive heart failure following bilateral lung transplant in whom pulmonary vagal afferent nerve activity was absent. PMID- 9727809 TI - Resolution of obstructive sleep apnoea with growth in the Robin sequence. AB - A 12 year old female with the Robin sequence presented with a one year history of snoring, witnessed apnoeas and daytime sleepiness. Surgery in early childhood had consisted of cleft palate repair, tonsillectomy and adenoidectomy and, later, revision palatoplasty. Overnight polysomnography (PSG) demonstrated severe obstructive sleep apnoea syndrome with an apnoea/hypopnoea index (AHI) of 49 events x h(-1), and repetitive oxygen desaturations below 50%. Nasal continuous positive airway pressure (nCPAP) effectively controlled her sleep abnormalities. After 3 yrs of nCPAP therapy, she requested discontinuation and was fully reassessed. PSG without nCPAP revealed an AHI <5 events x h(-1) with no desaturations below 90% and normal sleep quality. A repeat lateral cephalometrogram showed increased mandibular length and posterior airway space and reduced soft palate length. The patient remains asymptomatic 9 months following nCPAP discontinuation. This case indicates that nasal continuous positive airway pressure is an effective nonsurgical therapy in children with obstructive sleep apnoea syndrome and the Robin sequence. It is likely that mandibular growth, increase in mandibular length and enlargement of the posterior airway space was responsible for the resolution of obstructive sleep apnoea syndrome in this case. PMID- 9727810 TI - Eosinophilic lung disease associated with Candida albicans allergy. AB - A significant number of cases of chronic eosinophilic pneumonia remain idiopathic in spite of a comprehensive search of associated causes. This study reports a patient with a classical clinical presentation of chronic eosinophilic pneumonia and peripheral blood eosinophilia in whom selective sensitization to Candida albicans was demonstrated. This yeast was present in the bronchoalveolar lavage culture and specific serum immunoglobulin (Ig)E and IgG against C. albicans were found in the patient's serum. Levels of these specific immunoglobulins diminished with corticosteroid treatment and increased coinciding with a new outbreak of symptoms after lowering the dosage of corticosteroids. To the author's knowledge, this is the first case described of chronic eosinophilic pneumonia associated with sensitization to C. albicans. Evaluation of allergy to C. albicans should be performed in chronic eosinophilic pneumonia before labelling cases as idiopathic. PMID- 9727811 TI - Standardized tuberculosis treatment outcome monitoring in Europe. Recommendations of a Working Group of the World Health Organization (WHO) and the European Region of the International Union Against Tuberculosis and Lung Disease (IUATLD) for uniform reporting by cohort analysis of treatment outcome in tuberculosis patients. AB - Consensus-based recommendations have been developed by a Working Group of the World Health Organisation (WHO) and the International Union Against Tuberculosis and Lung Disease (IUATLD) on uniform reporting of tuberculosis (TB) treatment outcome data in countries in Europe. The main purpose of treatment monitoring is to find out how many of the potential infectious TB patients notified were declared cured at the end of treatment. Following the uniform case definitions as defined in 1996, emphasis is placed on cohort analysis of definite cases of pulmonary TB. The Working Group recommends using a minimal set of six mutually exclusive categories of treatment outcome: cure, treatment completed, failure, death, treatment interrupted, and transfer out. More detailed subsets may be chosen. Treatment outcome is expressed as a percentage of the total number of cases notified. Analysis should be separate for new and retreatment cases. Treatment outcome data have to be collected at the local level and passed on to regional and national authorities on an ongoing basis. Evaluation of treatment results becomes, preferably, an inbuilt component of national monitoring of programme performance. Because of the long duration of treatment, it is recommended that analysis is carried out in the first quarter of the calendar year that follows a full year after the last patient was enrolled. Feedback is essential. Treatment outcome results should become an inseparable part of the annual report on tuberculosis. PMID- 9727812 TI - Eleven peak flow meters: a clinical evaluation. PMID- 9727813 TI - The impact of human immunodeficiency virus disease on academic health centers. PMID- 9727814 TI - Prospective evaluation of an index for predicting the risk of major bleeding in outpatients treated with warfarin. AB - PURPOSE: To evaluate the accuracy and clinical utility of the Outpatient Bleeding Risk Index for estimating the probability of major bleeding in outpatients treated with warfarin. The index was previously derived in a retrospective cohort of 556 patients from a different hospital (derivation cohort). SUBJECTS AND METHODS: We enrolled 264 outpatients starting warfarin (validation cohort) to validate the index prospectively. All patients were identified upon hospital discharge, and physician estimates of the probability of major bleeding were obtained before discharge in the validation cohort. RESULTS: Major bleeding occurred in 87 of 820 outpatients (6.5%/yr). The index included four independent risk factors for major bleeding: age 65 years or greater; history of gastrointestinal bleeding; history of stroke; and one or more of four specific comorbid conditions. In the validation cohort, the index predicted major bleeding: the cumulative incidence at 48 months was 3% in 80 low-risk patients, 12% in 166 intermediate-risk patients, and 53% in 18 high-risk patients (c index, 0.78). The index performed better than physicians, who estimated the probability of major bleeding no better than expected by chance. Of the 18 episodes of major bleeding that occurred in high-risk patients, 17 were potentially preventable. CONCLUSIONS: The Outpatient Bleeding Risk Index prospectively classified patients according to risk of major bleeding and performed better than physicians. Major bleeding may be preventable in many high-risk patients by avoidance of over anticoagulation and nonsteroidal anti-inflammatory agents. PMID- 9727815 TI - A randomized trial of buprenorphine maintenance for heroin dependence in a primary care clinic for substance users versus a methadone clinic. AB - PURPOSE: Buprenorphine is an alternative to methadone for the maintenance treatment of heroine dependence and may be effective on a thrice weekly basis. Our objective was to evaluate the effect of thrice weekly buprenorphine maintenance for the treatment of heroin dependence in a primary care clinic on retention in treatment and illicit opioid use. SUBJECTS AND METHODS: Opioid dependent patients were randomly assigned to receive thrice weekly buprenorphine maintenance in a primary care clinic that was affiliated with a drug treatment program (n = 23) or in a traditional drug treatment program (n = 23) in a 12-week clinical trial. Primary outcomes were retention in treatment and urine toxicology for opioids; secondary outcomes were opioid withdrawal symptoms and toxicology for cocaine. RESULTS: Retention during the 12-week study was higher in the primary care setting (78%, 18 of 23) than in the drug treatment setting (52%, 12 of 23; P = 0.06). Patients admitted to primary care had lower rates of opioid use based on overall urine toxicology (63% versus 85%, P < 0.01) and were more likely to achieve 3 or more consecutive weeks of abstinence (43% versus 13%, P = 0.02). Cocaine use was similar in both settings. CONCLUSIONS: Buprenorphine maintenance is an effective treatment for heroin dependence in a primary care setting. PMID- 9727816 TI - Should pericardial drainage be performed routinely in patients who have a large pericardial effusion without tamponade? AB - PURPOSE: To assess whether drainage of pericardial effusion by pericardiocentesis or surgery is justified as a routine measure in the initial management of patients with large pericardial effusion without tamponade or suspected purulent pericarditis. SUBJECTS AND METHODS: All patients with large pericardial effusion without tamponade or suspected purulent pericarditis who were seen at our institution during a span of 6 years (1990 to 1995) were retrospectively (46) or prospectively (25) reviewed. Large pericardial effusion was defined as a sum of echo-free pericardial spaces in diastole exceeding 20 mm. RESULTS: Large pericardial effusion was diagnosed in 162 patients, 71 of whom fulfilled criteria for inclusion. Of these, 26 underwent a pericardial drainage procedure. Diagnostic yield was 7%, as only 2 specific diagnoses were made using these procedures. During follow-up (95% of patients, median 10 months), no patient developed cardiac tamponade or died as a result of pericardial disease, nor did any new diagnoses become manifest in the 45 patients who did not have pericardial drainage initially. Moderate or large effusions persisted in only 2 of 45 patients managed conservatively. CONCLUSIONS: Routine pericardial drainage procedures have a very low diagnostic yield in patients with large pericardial effusion without tamponade or suspected purulent pericarditis, and no clear therapeutic benefit is obtained with this approach. Clinical outcomes depend on underlying diseases, and do not appear to be influenced by drainage of pericardial fluid. PMID- 9727819 TI - Variation in the mineral content of commercially available bottled waters: implications for health and disease. AB - PURPOSE: Although the annual consumption of bottled water in North America is 12.7 gallons per capita, little is known about the potential health effects of these waters. We reviewed the amounts of major minerals found in commercially available bottled waters, the recommended daily allowances for these minerals, and their beneficial and harmful effects. METHODS: We obtained the mineral content of various commercially available bottled waters in North America and Europe from The Pocket Guide to Bottled Water. We then conducted a Medline search to identify articles examining the beneficial and harmful effects of magnesium, sodium, and calcium. RESULTS: Great variation exists in the mineral content of commercially available bottled waters. Among the bottled waters that we reviewed, the magnesium content ranges from 0 to 126 mg per liter, the sodium content ranges from 0 to 1,200 mg per liter, and the calcium content ranges from 0 to 546 mg per liter. Epidemiologic and clinical studies suggest that magnesium may reduce the frequency of sudden death, that sodium contributes to the occurrence of hypertension, and that calcium may help prevent osteoporosis. CONCLUSION: The ideal bottled water should be rich in magnesium and calcium and have a low sodium content. Because there is great variation in the mineral content of commercially available bottled waters, the actual mineral content of bottled water should be considered when selecting one for consumption. PMID- 9727818 TI - Contrasting actions of pressor agents in severe autonomic failure. AB - BACKGROUND: Orthostatic hypotension is the most disabling symptom of autonomic failure. The choice of a pressor agent is largely empiric, and it would be of great value to define predictors of a response. PATIENTS AND METHODS: In 35 patients with severe orthostatic hypotension due to multiple system atrophy or pure autonomic failure, we determined the effect on seated systolic blood pressure (SBP) of placebo, phenylpropanolamine (12.5 mg and 25 mg), yohimbine (5.4 mg), indomethacin (50 mg), ibuprofen (600 mg), caffeine (250 mg), and methylphenidate (5 mg). In a subgroup of patients, we compared the pressor effect of midodrine (5 mg) with the effect of phenylpropanolamine (12.5 mg). RESULTS: There were no significant differences in the pressor responses between patients with multiple system atrophy or pure autonomic failure. When compared with placebo, the pressor response was significant for phenylpropanolamine, yohimbine, and indomethacin. In a subgroup of patients, we confirmed that this pressor effect of phenylpropanolamine, yohimbine, and indomethacin corresponded to a significant increase in standing SBP. The pressor responses to ibuprofen, caffeine, and methylphenidate were not significantly different from placebo. Phenylpropanolamine and midodrine elicited similar pressor responses. There were no significant associations between drug response and autonomic function testing, postprandial hypotension, or plasma catecholamine levels. CONCLUSIONS: We conclude that significant increases in systolic blood pressure can be obtained in patients with orthostatic hypotension due to primary autonomic failure with phenylpropanolamine in low doses or yohimbine or indomethacin in moderate doses. The response to a pressor agent cannot be predicted by autonomic function testing or plasma catecholamines. Therefore, empiric testing with a sequence of medications, based on the risk of side effects in the individual patient and the probability of a response, is a useful approach. PMID- 9727820 TI - Taking stock at The Green Journal. PMID- 9727817 TI - A case series of hospitalized patients with elevated digoxin levels. AB - PURPOSE: Although there is renewed enthusiasm for the use of digoxin in patients with heart failure, current dosing guidelines are based on a nomogram published in 1974. We studied the incidence of and risk factors for elevated digoxin levels in patients admitted to a community hospital, and compared their dosage regimens to published guidelines. SUBJECTS AND METHODS: We reviewed the charts of all patients who had serum digoxin levels greater than 2.4 ng/mL during a 6-month period. We collected demographic and clinical data, indications for digoxin use, digoxin dosage, concurrent medications, laboratory data, and clinical and electrocardiographic features of digoxin toxicity. RESULTS: Of the 1,433 patients with digoxin assays, 115 (8%) patients had elevated levels. Of the 82 patients with complete records and correctly timed digoxin levels, 59 (72%) had electrocardiographic or clinical features of digoxin toxicity. Patients with serum digoxin levels >2.4 ng/mL were slightly older (78 +/- 8 versus 73 +/- 9 years of age; P = 0.12) and had greater serum creatinine levels (3.1 +/- 7.3 versus 1.4 +/- 0.3 mg/dL; P = 0.01) than those with levels < or =2.4 ng/mL. Forty seven patients had elevated digoxin levels on admission, including 21 patients admitted for digoxin toxicity. Impaired or worsening renal function contributed to high levels in 37 patients, and a drug interaction was a contributory factor in 10 cases. Twenty (43%) of these patients were taking the recommended maintenance dose based on the scheme employed in the Digitalis Investigation Group study. Thirty-five patients developed high digoxin levels while in hospital. In 26 patients, this followed a loading dose of digoxin for the control of rapid atrial fibrillation. Impaired renal function was implicated in all of these patients. Despite the elevated digoxin level, rate control was achieved in only 11 patients of these patients. CONCLUSIONS: Elevated digoxin levels and clinical toxicity remains a common adverse drug reaction. Elderly patients, particularly those with impaired renal function and low body weights, are at the greatest risk. As published digoxin nomograms often result in toxicity, clinical variables need to be monitored. In patients with congestive heart failure and normal sinus rhythm the potential benefit of digoxin is small; thus, patients should receive a dose that minimizes the risk of toxicity. For patients with new onset atrial fibrillation, other agents may be preferable for rate control. PMID- 9727821 TI - New insights into erectile dysfunction: a practical approach. AB - Erectile dysfunction (ED) is the most common sexual problem in men, after premature ejaculation, affecting up to 30 million in the United States. In a society in which sexuality is widely promoted, ED impacts on feelings of self worth and self-confidence and may impair the quality of life of affected men and their partners. Damage to personal relationships can ensue; and the anger, depression, and anxiety engendered spill over into all aspects of life. Patients are often embarrassed or reluctant to discuss the matter with their primary care practitioners. Unfortunately, many physicians fail to take the opportunity to promote open discussion of sexual dysfunction. They too, may avoid the topic through personal embarrassment. Since the National Institutes of Health (NIH) Consensus Conference on Impotence in 1992, the inadequate level of public and professional understanding of ED has begun to be addressed. As a first step in breaking down the communication barriers between patients and practitioners, it is important that physicians have a thorough understanding of the wide variety of conditions associated with ED and how the different risk factors for ED may be readily identified. This review addresses the diagnosis of ED and identifies diagnostic tests that can be used by primary care physicians to determine the patients most at risk and the treatments most suited to meet the patients' and their partners' goal for therapy. PMID- 9727822 TI - The etiology of obesity: relative contribution of metabolic factors, diet, and physical activity. AB - Three major factors modulate body weight: metabolic factors, diet, and physical activity, each influenced by genetic traits. Despite recent advances in these areas, the prevalence of obesity in Westernized societies has increased. In contrast to monogenic animal models and rare human genetic syndromes, predisposition to common forms of obesity is probably influenced by numerous susceptibility genes, accounting for variations in energy requirements, fuel utilization, muscle metabolic characteristics, and taste preferences. Although recent increases in obesity prevalence cannot be explained by changes in the gene pool, previously "silent" genetic variants may now play important permissive roles in modern societies. Available data suggest that variations in resting energy expenditure, thermic effect of food, and fuel utilization exist but, by themselves, are unlikely to explain the onset of obesity. Regarding diet, the best available trend survey data indicate that fat and energy intake have fallen, in this and other Westernized countries. Diverging trends of decreasing energy intake and increasing body weight suggest that reduced physical activity may be the most important current factor explaining the rising prevalence of obesity. Subsistence in modern societies requires extreme adaptations in previously useful energy-conserving diet and exercise behaviors. Recognizing the difficulties in sustaining energy-restricted diets in the presence of fast foods and social feasts, the current trend toward increasing body weight is not likely to be reversed solely through recommendations for further reductions in energy intake. In all likelihood, activity levels will have to increase in response to an environment engineered to be more physically demanding. PMID- 9727823 TI - Bartter syndrome: unraveling the pathophysiologic enigma. AB - Familial hypokalemic, hypochloremic metabolic alkalosis, or Bartter syndrome, is not a single disorder but rather a set of closely related disorders. These Bartter-like syndromes share many of the same physiologic derangements, but differ with regard to the age of onset, the presenting symptoms, the magnitude of urinary potassium (K) and prostaglandin excretion, and the extent of urinary calcium excretion. At least three clinical phenotypes have been distinguished: (1) classic Bartter syndrome; (2) the hypocalciuric-hypomagnesemic Gitelman variant; and (3) the antenatal hypercalciuric variant (also termed hyperprostaglandin E syndrome). The fundamental pathogenesis of this complex set of disorders has long fascinated and stymied investigators. Physiologic investigations have suggested numerous pathogenic models. The cloning of genes encoding renal transport proteins has provided molecular tools to begin testing these hypotheses. To date, molecular genetic analyses have determined that mutations in the gene encoding the thiazide-sensitive sodium-chloride (Na-Cl) cotransporter underlie the pathogenesis of the Gitelman variant. In comparison, the antenatal variant is genetically heterogeneous with mutations in the genes encoding either the bumetanide-sensitive sodium-potassium-chloride (Na-K-2Cl) cotransporter or the luminal, ATP-regulated, K channel. With these data, investigators have begun to unravel the pathophysiologic enigma of the Bartter like syndromes. Further studies will help refine pathogenic models for this set of disorders as well as provide new insights into the normal mechanisms of renal electrolyte transport. PMID- 9727824 TI - Effects of allogeneic peripheral stem cell transplantation in a patient with job syndrome of hyperimmunoglobulinemia E and recurrent infections. PMID- 9727826 TI - The Green Journal, then and now. PMID- 9727825 TI - Methicillin-resistant Staphylococcal enterocolitis. PMID- 9727828 TI - An uncommon pain in the neck. PMID- 9727827 TI - Clinical trials in departments of internal medicine. PMID- 9727829 TI - The -4 kilobase promoter region of the winged helix transcription factor HNF 3alpha gene elicits transgene expression in mouse embryonic hepatic and intestinal diverticula. AB - Murine hepatocyte nuclear factor-3alpha (HNF-3alpha) protein is a member of a large family of developmentally regulated transcription factors that share homology in the winged helix/fork head DNA binding domain and participate in embryonic pattern formation. HNF-3alpha also mediates cell-specific transcription of genes important for the function of hepatocytes, intestinal, pancreatic and bronchiolar epithelium. We have previously determined that -520 nucleotides upstream of the rat HNF-3alpha gene were sufficient to elicit hepatoma-specific expression in transfection assays and reported on a novel HNF-3alpha expression pattern in the renal pelvis urothelium of the embryonic and adult kidney. We also showed that retinoic acid mediated activation of the HNF-3alpha gene required -4 kb of the HNF-3alpha promoter region in F9 teratocarcinoma transfections. In order to determine regulatory sequences mediating the HNF-3alpha cellular expression pattern in developing mouse embryos, we created transgenic mice bearing the -4 kb HNF-3alpha promoter region driving expression of the beta galactosidase transgene. Embryonic analysis of two transgenic mouse lines demonstrated that the -4 kb HNF-3alpha promoter sequences were sufficient to elicit transgene expression in the developing liver, intestine, esophagus, nasal epithelial cells and floorplate of the neurotube, but not in the mesodermal notochord or in the lung bud. One of the transgenic lines also exhibited proper expression in the mesonephric ducts and metanephric diverticulum, suggesting that the -4 kb HNF-3alpha promoter region contained a subset of the regulatory sequences necessary for HNF-3alpha expression in the developing kidney. PMID- 9727830 TI - X-twi is expressed prior to gastrulation in presumptive neurectodermal and mesodermal cells in dorsalized and ventralized Xenopus laevis embryos. AB - Early X-twi expression has been now investigated from egg laying to the early neurulation stages in Xenopus embryos, using both in situ hydridization and the more sensitive techniques of RT-PCR. We show that in unfertilized eggs, a decreasing gradient of X-twi transcript distribution is observed from animal to vegetative caps. X-twi RNA can be weakly detected at stages prior to gastrulation, and with increased intensity from stage 8 onwards. At blastula, X twi transcripts are located towards the animal pole, and as gastrulation begins, they are detected in the developing axial mesoderm and then they accumulate in the sensorial layer of the neurectoderm, the mesodermal layer and in neural crest cells up to late neurula stages. We show, in addition, that in lithium-chloride- and UV-treated Xenopus embryos (that are respectively both "anteriorized/dorsalized" and in "posteriorized/ventralized"), X-twi RNA is detected in cells in similar positions to those that express X-twi in normal embryos. As a whole, our results show that X-twi is expressed even when regionalization of the mesoderm is disturbed and raises the question of a putative function of X-twi prior to gastrulation. PMID- 9727831 TI - The expression of XIF3 in undifferentiated anterior neuroectoderm, but not in primary neurons, is induced by the neuralizing agent noggin. AB - The gene XIF3 encodes a neural-specific type-III intermediate filament protein whose expression in the embryo precedes that of the neurofilaments by several hours. We now show, by in situ hybridization, that it is expressed at the neurula stage in primary neurons and, to a lesser extent, in undifferentiated anterior neuroectoderm. At the swimming tadpole stage, strong expression is restricted to the midbrain-hindbrain boundary, even-numbered rhombomeres of the hindbrain and the Vth and VIIth cranial ganglia. XIF3 gene expression can be induced in ectodermal cells (animal caps) derived from blastula when grown to the neurula stage in the presence of the neuralizing agent noggin. In agreement with the proposed ability of noggin to neuralize, but not to promote neuronal differentiation, we find that the pattern of noggin-inducible XIF3 expression in animal caps is consistent with expression in undifferentiated anterior neuroectoderm but not in primary neurons. PMID- 9727833 TI - HMG-17, a chromosomal non-histone protein, shows developmental regulation during organogenesis. AB - We used the differential hybridization technique for isolating developmentally regulated genes from the mouse metanephric kidney. In this screening, we identified the cDNA encoding high-mobility-group protein 17 (HMG-17), a chromosomal non-histone protein which modulates the conformation of transcriptionally active chromatin. Using Northern blot analysis, the HMG-17 mRNA was strongly expressed during embryogenesis and downregulated in various adult murine organs. At the histological level, the transcript localized to differentiating tissue regions and was apparently downregulated in mature structures indicating that HMG-17 expression is linked to cell differentiation. HMG-17 can thus be regarded as a general marker for tissues or cells undergoing differentiation during organogenesis. PMID- 9727832 TI - Evidence for non-axial A/P patterning in the nonneural ectoderm of Xenopus and zebrafish pregastrula embryos. AB - Recent studies in early Xenopus and zebrafish embryos have demonstrated that posteriorizing, non-axial signals arising from outside the organizer (or shield) contribute to A/P patterning of the neural axis, in contradiction to the classical Spemann model in which such signals were proposed to be solely organizer derived. Our studies on the early expression of the transcription factors GATA-2 and 3 in both Xenopus and zebrafish nonneural ectoderm lend support to the existence of such non-axial signaling in the A/P axis. Thus we find that the earliest expression of GATA-2 and 3 is located in nonneural ectoderm and is strongly patterned in a graded manner along the A/P axis, being high anteriorly and absent from the most posterior regions. This results by early neurula stages in three broad zones: an anterior region which is positive for both GATA-2 and 3, a middle region which is positive for GATA-2 alone and a posterior region in which neither gene is expressed. These regions correspond to head, trunk and tail ectoderm and may represent the beginnings of functional segmentation of nonneural ectoderm, as suggested in the concept of the 'ectomere'. We find that A/P patterning of GATA expression in nonneural ectoderm may occur as early as late blastula/early gastrula stages. We investigate which posteriorizing signals might contribute to such distinct non axial ectodermal patterning in the A/P axis and provide evidence that both FGF and a Wnt family member contribute towards the final A/P pattern of GATA expression in nonneural ectoderm. PMID- 9727835 TI - Expression of galectin-1 in the mouse olfactory system. AB - Primary sensory olfactory axons arise from the olfactory neuroepithelium that lines the nasal cavity and then project via the olfactory nerve into the olfactory bulb. The beta-galactoside binding lectin, galectin-1, and its laminin ligand have been implicated in the growth of these axons along this pathway. In galectin-1 null mutant mice, a subpopulation of primary sensory olfactory axons fails to reach its targets in the olfactory bulb. In the present study we examined the spatiotemporal expression pattern of galectin-1 in normal mice in order to understand its role in the development of the olfactory nerve pathway. At E15.5, when olfactory axons have already contacted the olfactory bulb, galectin-1 was expressed in the cartilage and mesenchyme surrounding the nasal cavity but was absent from the olfactory neuroepithelium, nerve and bulb. Between E16.5 and birth galectin-1 began to be expressed by olfactory nerve ensheathing cells in the lamina propria of the neuroepithelium and nerve fibre layer. Galectin-1 was neither expressed by primary sensory neurons in the olfactory neuroepithelium nor by their axons in the olfactory nerve. Laminin, a galectin-1 ligand, also exhibited a similar expression pattern in the embryonic olfactory nerve pathway. Our results reveal that galectin-1 is dynamically expressed by glial elements within the nerve fibre layer during a discrete period in the developing olfactory nerve pathway. Previous studies have reported galectin-1 acts as a substrate adhesion molecule by cross-linking primary sensory olfactory neurons to laminin. Thus, the coordinate expression of galectin-1 and laminin in the embryonic nerve fibre layer suggests that these molecules support the adhesion and fasciculation of axons en route to their glomerular targets. PMID- 9727834 TI - Trefoil peptides are early markers of gastrointestinal maturation in the rat. AB - Trefoil peptides are members of a unique family of proteins found predominately throughout the gastrointestinal tract, whose proposed functions include mucus stabilization, stimulation and/or differentiation of epithelial cells during wound repair. Recent trefoil knockout studies have reported delays in epithelial cell migration or maturation pathways together with almost a complete lack of mucus. In order to fully explore the role of trefoil peptides in gastrointestinal maturation, these studies were undertaken to accurately characterize the expression of trefoil peptides in the developing rat gut. The results of RPA suggest that trefoil mRNA's are expressed as early as 15 days post coitus (dpc) in the intestine and stomach. Proteins are detected at 17 dpc by radioimmunoassay and immunohistochemical studies, which localize trefoil peptide expression to the lumenal surface of epithelial cells. At 17 dpc the gut is lined by pseudo stratified, undifferentiated epithelial cells. Polarized, columnar cells are not detected until at least 18 dpc, with sparse mucus staining and parietal cell markers not being detected until 18 and 19 dpc respectively. This data demonstrates that trefoil peptides are early markers of epithelial cell maturation in the developing rat gut. The time course of their expression, well before the mucus cell type is specified, suggests a potential role in epithelial cell differentiation. PMID- 9727836 TI - The pattern of Distal-less expression in the mouthparts of crustaceans, myriapods and insects: new evidence for a gnathobasic mandible and the common origin of Mandibulata. AB - We examined embryos of representatives of crustaceans, myriapods and insects with respect to DII expression in the mouthparts. In order to examine the relationships between mandibular DII expression and the occurrence of a mandibular palp we compared amphipod, isopod and decapod crustacean species. In species with mandibular palps, DII expression is maintained throughout development and is restricted to the palps. The species lacking a palp as an adult show only transient DII expression in early embryonic stages. Furthermore, we studied mandibular DII expression in the myriapod Glomeris marginata that lacks like all myriapods mandibular palps as an adult. The expression pattern is similar to that in crustaceans lacking a palp as an adult. We examined entognathous and ectognathous insects. No sign of mandibular expression could be detected. It is shown that the distal parts of the mandibular appendage were reduced in several steps and lineages independently up to a total loss. Furthermore, we studied DII expression in the first and second maxillae. Except for Glomeris and the collembolans, the first maxillae of all species show a similar pattern of three lobes expressing DII: the outer expression marks the maxillary palp and the inner two mark the outgrowing endites (galea and lacinia of insects). In the first maxillae of collembolans only two expression areas could be detected. In palpless adult first maxillae of isopod crustaceans a transitory embryonic palp occurs which is also DII positive. In the second maxillae of insects, isopod and amphipod crustaceans only two DII-positive lobes occur. Our data suggest a gnathobasic character of the mandibles of crustaceans, myriapods and insects supporting the monophyly of Mandibulata sensu Snodgrass. The interpretation of DII expression patterns and its limits are critically evaluated. PMID- 9727837 TI - P450scc-like immunoreactivity throughout gonadal restructuring in the protogynous hermaphrodite Thalassoma duperrey. AB - The source of steroid hormones, which potentially regulate gonadal restructuring throughout protogynous sex change in teleosts, remains largely unknown. To address this issue, immunocytochemical methods were employed to detect gonadal sites of steroidogenesis in the protogynous hermaphrodite wrasse Thalassoma duperrey at different stages in the sex change process. Steroidogenic cells were classified based on the presence of P450 cholesterol-side-chain-cleavage-like immunoreactivity (P450scc-ir). P450scc-ir cells were predominantly in the thecal layer of normal females. As females underwent sex change, P450scc-ir localization shifted from the thecal layer to the interstitium. P450scc-ir cells appeared to increase in number midway through sex change. In sex-changed males, P450scc-ir cells were found in small clusters interspersed among spermatogenic lobules. These results demonstrate for the first time the ability of the gonad to produce potential steroidal mediators of gonadal restructuring throughout the sex change process. PMID- 9727838 TI - Immunohistochemical localization of TGF-beta type II receptor and TGF-beta3 during palatogenesis in vivo and in vitro. AB - The disappearance of medial edge epithelium (MEE) is a critical event for palate fusion. TGF-beta3 is one factor participating in the regulation of this process. To investigate the nature of ligand-receptor interactions in vivo between TGF beta3 and the type II TGF-beta receptor (TbetaR-II), we compared the expression pattern of the receptor with TGF-beta3. Immunohistochemical analysis of the mouse fetus from E12 to E15 showed that expression of TbetaR-II in the palate began at E13 when the palatal shelves were in a vertical orientation. TbetaR-II was localized in the epithelial cells. This epithelium-favored distribution remained during palatal shelf elevation, the medial edge epithelial adherence, and midline epithelial seam disruption. After palate fusion and mesenchyme confluence, weak expression of TbetaR-II was present in the mesenchyme. To verify the possibility that TGF-beta3 and TbetaR-II expression coincide, immunohistochemistry was used to localize them both in serial sections. The distribution pattern of TGF-beta3 was also epithelium-limited in the palate from E13 to E15, and the spatial localization was correlated with the expression of TbetaR-II. Immunohistochemical localization of TbetaR-II and TGF-beta3 in palatal shelves in organ culture had patterns that were consistent with the in vivo results. These results suggest that TGF-beta3 exerts its developmental role through TbetaR-II in an autocrine fashion. The expression of both TGF-beta3 and TbetaR-II was below the detectable level in the mesenchyme following MEE disruption, suggesting that the TGF-beta3 signal might not be required once the MEE has completed phenotypic transformation/migration. PMID- 9727839 TI - Hormonal factors from the mammalian pineal gland interfere with cell development in Hydra. AB - A partially purified, melatonin-free low-molecular-weight extract from the ovine pineal gland with antitumor activity (YC05R), interferes with terminal differentiation in the interstitial cell line of Hydra. Nematoblasts developed into defective nematocytes that were subject to cell death and the tentacles eventually became devoid of nematocytes. In an attempt to identify the causative components of the extract, several known potential constituents were assayed. Two factors were found to have similar effects, although only in rather high concentrations: 1alpha, 25 dihydroxyvitamin D3 (>150 nM) and pinoline (>5 microM), a natural tryptophan-derived beta-carboline. The proliferative activity in the interstitial cell line was only slightly reduced by these factors. Two other beta-carbolines that occur in the mammalian brain, harmine (10 microM) and n-butyl-beta-carboline-3-carboxylate (beta-CCB), caused the premature death of epithelial cells and thus the development of dwarfish animals which, however, continued to generate new animals by budding. The pineal extract probably contains some more, still unidentified components that interfere more potently with cell development, in Hydra as well as in mammals. PMID- 9727841 TI - Reverse mutations--spontaneous amelioration or cure of inherited disorders? AB - Some recent publications indicate that inherited disorders can ameliorate or possibly disappear if mutations responsible for the disease revert to normal. This review tries to summarize our current knowledge about reverse mutations as this information may be of special interest for attempts at somatic gene therapy. PMID- 9727840 TI - Low-molecular-weight hormonal factors that affect head formation in Hydra. AB - Support or inhibition of DAG-induced head formation: Hydra magnipapillata can be caused to form ectopic head structures by periodictreatmentwith PKC activators such as diacylglycerol (DAG). This ectopic head formation is supported by an extract from the ovine pineal gland that contains low-molecular-weight compounds. The frequency of ectopic head formation is also increased when DAG is paired with one of several lipophilic hormonal factors: (1) pinoline, a putative pineal hormone derived from tryptophan, (2) 12-S-HETE, a paracrine derivative of arachidonic acid, or (3) 1alpha, 25 dihydroxyvitamine D3, a hormonal factor also known as calcitriol. Dose-response curves were non-monotonic passing a maximum at low dosages. By contrast, DAG lost its capacity to induce ectopic head structures when it was paired with the provitamin D3, cholecalciferol. Patterning the head: one eicosanoid was found which influences patterning without being combined with DAG: 12-R-HETE caused growth-based elongation of the tentacles and an increase in the number of tentacles without affecting the longitudinal body pattern. Similar effects are brought about by substances that interfere with tyrosine phosphorylation, most potently bythe phosphotyrosine phosphatase inhibitor bisperoxo-(1,10-phenanthroline)-oxovanadate (V). The results speak for the existence of head-inducing hormonal factors, underline the significance of content protein kinases to pattern formation and point to a negative influence of the vitamin D3 content of the food on the capacity of the animals to develop head structures. PMID- 9727843 TI - Ruby laser therapy for labial lentigines in Peutz-Jeghers syndrome. AB - Some patients with Peutz-Jeghers syndrome may be disturbed by the appearance of lentigines. Such patients require management of their lentigines as well as their gastro-intestinal polyps. We describe ruby laser therapy of labial lentigines in two children with Peutz-Jeghers syndrome. The response to treatment was excellent and no sequelae or recurrence of the lesions was noted. CONCLUSION: Our experience suggests that ruby laser therapy is safe and a suitable approach for the treatment of labial lentigines in children with Peutz-Jeghers syndrome. PMID- 9727842 TI - Amlodipine once-daily in systemic hypertension. AB - The calcium channel blocker nifedipine is widely used in children with systemic hypertension: however, because of the short duration of action, three to four daily doses of the standard preparation are required. Amlodipine once-daily, a calcium channel blocker structurally related to nifedipine with an excellent bioavailability and a long elimination half-time, has been shown to reduce blood pressure in adults. No information is available on the use of amlodipine in childhood. The effects of amlodipine once-daily (5 to 10 mg) were therefore assessed in 28 paediatric patients with hypertension. Amlodipine was withdrawn in five patients who experienced oedema and flushing. In the remaining 23 patients blood pressure was significantly reduced 3 weeks after amlodipine (on average by 7/5 mm Hg) and further decreased at 12 weeks (by 21/12 mm Hg). Heart rate and body weight were unchanged. In eight patients concomitantly treated with cyclosporine, the blood level of this agent was stable throughout the study, thus not requiring any dose adjustment. CONCLUSION: The study illustrates the antihypertensive properties of amlodipine once-daily in paediatric hypertension. Amlodipine appears particularly indicated in patients concomitantly treated with cyclosporine. PMID- 9727844 TI - Experience of severe hypoglycaemia may influence both patient's and physician's subsequent treatment policy of insulin-dependent diabetes mellitus. AB - Daily insulin doses and HbA1c were studied 0-3 months before and 2-6, 7-11, and 12-16 months after 48 consecutive episodes of severe hypoglycaemia (coma and/or convulsion) in children and adolescents with insulin-dependent diabetes mellitus. After 69% of the attacks, either physicians or patients or both decreased daily insulin doses (for the whole group, mean SD: 0-3 months before the episode 0.93 0.20 U/kg vs 2-6 months after 0.84 0.20 U/kg, P < 0.001), which may have worsened the subsequent glycaemic control as evidenced by a significant increase in HbA1c (8.3+/-1.5% vs 9.1+/-1.8%, P< 0.001, respectively). Physicians decreased the insulin dose even in 14 of the 33 patients with a preventable cause for their hypoglycaemia other than erroneous excess of insulin. CONCLUSION: Experience of severe hypoglycaemia may worsen the subsequent glycaemic control. This might in part be due to an excessive lowering of daily insulin doses by both physicians as well as patients and their families. Hypoglycaemia management must include intensive education about prevention without compromising diabetes control. PMID- 9727846 TI - Reference values for height and weight in Prader-Willi syndrome based on 315 patients. AB - The spontaneous growth of 315 patients (109 girls and 208 boys) with Prader-Willi syndrome (PWS) was analysed in a mixed longitudinal and cross-sectional manner. 33 patients were seen in the department between 1970 and 1994; height and weight of 76 patients from Germany were evaluated by means of a questionnaire with detailed measuring instructions, and 206 definite cases were added from the literature. Mean ( SD) length of newborn babies with PWS was 50.2+/-2.8 cm (145 boys) and 48.9 3.3 cm (79 girls). Mean weight at birth was 2945 570 g in boys and 2782+/-594 g in girls. During the 1st year, the children's growth was nearly normal, thereafter short stature was present in approximately 50% of PWS patients. Between 3 and 13 years of age, the 50th percentile for height in PWS is roughly identical with the 3rd percentile in healthy controls. Body weight was normal for all boys and girls during the first 2 years. Thereafter, a rapid weight gain occurred; after an age of 10 years weight-for-height index in nearly all patients exceeded the normal range. The extent of pubertal growth was reduced for the group. Mean adult height was 161.6+/-8.1 cm (23 males) and 150.2+/-5.5 cm (21 females). Head circumference for age was normal for boys and girls. CONCLUSION: Reference data on spontaneous development of growth and weight gain of children with Prader-Willi syndrome are described allowing a better counselling of patients and parents. PMID- 9727845 TI - Long-term treatment of persistent hyperinsulinaemic hypoglycaemia of infancy with diazoxide: a retrospective review of 77 cases and analysis of efficacy-predicting criteria. AB - Primary persistent hyperinsulinaemic hypoglycaemia of infancy is rare. Diazoxide treatment remains the mainstay of medical therapy in long-term management. We reviewed 77 cases of primary persistent hyperinsulinism in neonates and infants who were treated with diazoxide and studied criteria predictive of therapeutic efficacy. The only criterion identified was age at manifestation. All but 1 of the 31 neonatal cases were unresponsive to diazoxide. Responsiveness increased with age: 12 of 39 early-infantile cases, and all seven late-infantile cases were diazoxide-responsive. In responders, a diazoxide dose of 10-15 mg/kg per day was always effective, suggesting an "all or none" response. Diazoxide-resistant hyperinsulinism is characterized by its severity with higher plasma insulin levels. The analysis of 46 surgically treated patients showed that the efficacy of diazoxide is not related to the aetiology of the pancreatic lesions. In six cases, after many years of management, diazoxide treatment was stopped without recurrence of hypoglycaemia. CONCLUSION: Diazoxide is an efficient treatment in the long-term management of most persistent hyperinsulinaemic hypoglycaemia of infancy revealed in infants and children but is usually ineffective in neonatal forms. Drug efficacy does not correlate with anatomical lesions. Medical treatment can sometimes be stopped after many years of management without recurrence of disease manifestations. PMID- 9727847 TI - Mitochondrial DNA deletion with Kearns Sayre syndrome in a child with Addison disease. AB - Kearns Sayre syndrome (KSS) is a multisystem disorder with a confounding variety of clinical manifestations, including ocular myopathy, pigmentary retinopathy, heart block and ataxia. Endocrinopathies are common in KSS, including growth hormone deficiency, hypogonadism, diabetes mellitus and hypoparathyroidism. A variety of deletions of mitochondrial DNA (mtDNA) are found in most cases. We report on a 5-year-old boy with Addison disease in whom further investigation revealed a 4.9 kilobase mtDNA deletion and KSS. Later he developed severe lactic acidosis and expired. CONCLUSION: The degree of mutant mtDNA heteroplasmy in various tissues on autopsy did not correlate well with the clinical manifestations, although this may be due at least in part to replacement with other tissue types. Our report is the first of non-autoimmune Addison disease in KSS and patients with KSS should be evaluated for adrenal insufficiency. Early recognition of adrenal insufficiency is crucial to prevent mortality from this cause. PMID- 9727848 TI - Thiamine-responsive lactic acidaemia: role of pyruvate dehydrogenase complex. AB - Lactic acidaemia is sometimes associated with a defect of the pyruvate dehydrogenase complex (PDHC), catalysing the thiamine-dependent decarboxylation of pyruvate. The activity of PDHC for different thiamine pyrophosphate (TPP) concentrations was determined in 13 patients with lactic acidaemia, clinically responsive to thiamine treatment in order to assess the role of PDHC in the aetiology of thiamine-responsive lactic acidaemia. Culture of lymphoblastoid cells and skin fibroblasts and muscle biopsies were performed in these 13 patients. The activity of PDHC to sodium dichloroacetate (DCA), known as the activator of PDHC, was also examined. Three groups were identified according to PDHC activity. Group 1 (two patients) displayed very low PDHC activity, which was not increased by DCA. This PDHC activity increased at high TPP concentrations. Group 2 (five patients) displayed below normal PDHC activity at low TPP concentrations, increased by DCA. This PDHC activity became normal at high TPP concentrations. PDHC deficiency in these patients of groups 1 and 2 was due to a decreased affinity of PDHC for TPP. Group 3 included six patients with normal PDHC activity at low as well as high TPP concentrations. This PDHC activity was increased by DCA. CONCLUSION: High concentrations of TPP may be required for maximal activity of PDHC in some patients with lactic acidaemia. The assay of PDHC activity, performed at a low concentration of TPP (1 x 10(-4)mM) allows selection of patients with thiamine-responsive lactic acidaemia. PMID- 9727849 TI - Cerebrospinal fluid shunts in the management of behavioural problems in Sanfilippo syndrome (MPS III). AB - Severe behavioural disturbance is a very common feature of Sanfilippo syndrome (mucopolysaccharidosis III, MPSIII), and one of the more difficult aspects of the disease to treat. We describe a series of six patients with MPS III who had cerebrospinal shunts inserted in an attempt to ameliorate behaviour that had proved refractory to conventional treatment. Symptoms improved significantly in all six but removal of the shunt was necessitated in one patient due to shunt blockage and infection. CONCLUSION: Our experience suggests cerebrospinal fluid shunting should be formally evaluated as an adjunct to conventional forms of treatment of extreme behavioural disturbance in MPS III. PMID- 9727850 TI - Doppler evaluation of renal blood flow velocity as a predictive index of acute renal failure in perinatal asphyxia. AB - Aim of our study was to evaluate Doppler renal blood flow velocity in asphyxiated neonates and to correlate renal function to Doppler findings. Doppler renal blood flow velocity was evaluated in 23 severely asphyxiated neonates born at a gestational age > 32 weeks and compared to our standard Doppler data obtained in 25 healthy neonates comparable for gestational age and birth weight. Renal Doppler ultrasound was performed on the 1st and 3rd days of life. Renal function was investigated in the first 2 weeks of life. Asphyxiated neonates showed mean values of systolic velocity and mean velocity significantly reduced (P < 0.001) compared with our standard Doppler values on the 1st day of life. Seven out of the 23 asphyxiated neonates were affected by acute renal failure and 14 showed no renal involvement. Two neonates were oliguric but did not develop acute renal failure. On the 1st day of life, neonates with acute renal failure had significantly lower mean values of systolic velocity and mean velocity than the asphyxiated neonates without renal involvement (P < 0.01). All 7 neonates affected by acute renal failure showed a systolic velocity more than 2SD below the mean standard value, while only 4 of the 16 asphyxiated neonates (25%) without acute renal failure had low systolic velocity values on the 1st day of life. Doppler velocities in asphyxiated neonates were similar to standard values on the 3rd day of life. Renal failure recovered before the 11th day of life in all cases. CONCLUSION: Our findings indicate that decreased Doppler renal flow systolic velocity observed in asphyxiated neonates on the st day of life is a useful predictive index for subsequent development of acute renal failure, with 100% sensitivity and 63.6% specificity. PMID- 9727851 TI - Pharmacokinetics of oral fluconazole in premature infants. AB - Systemic infections with Candida albicans in neonates are a frequent and well recognized problem. The therapeutic gold standard in this situation is the combined intravenous antimycotic treatment with amphotericin B and flucytosine. Potential adverse effects of this regimen have encouraged the search for desirable alternatives. We report on the use of oral fluconazole in neonates with Candida albicans septicaemia. Three premature infants were treated with four courses of therapy. Pharmacokinetic studies were performed during each course. At oral doses of 4.5-6 mg/kg once a day, serum levels of fluconazole were within the therapeutic range during the entire dosage interval. Follow up showed microbiological and clinical cure in all patients with no side-effects. In one patient a dosage of 4 mg/kg per day lead to a microbiological relapse with sub therapeutic serum levels. CONCLUSIONS: Oral fluconazole seems to be a safe and effective treatment for Candida albicans septicaemia even in premature infants. PMID- 9727853 TI - Clinical features of unilateral multicystic renal dysplasia in children. AB - A clinical study of 204 patients with unilateral multicystic renal dysplasia referred to 30 nephrology departments was undertaken to assess the frequency of complications in children who underwent nephrectomy (n = 40) versus those who were treated conservatively (n = 164). Six patients required antihypertensive treatment, 30 (13% of girls, 17% of boys) had at least one episode of urinary tract infection. The number of clinical complications did not differ in patients who underwent nephrectomy in comparison to those who did not. The dysplastic kidney decreased in size in 65% of kidneys with repeated ultrasound values; no change occurred in 16%, while an increase in maximal diameter was observed in 19%. Contralateral kidney length of more than 2 standard deviation scores (SDS) was seen in 30% of patients. There is evidence for a slight impairment of renal function in the whole study group given by a median of serum creatinine level of 0.63 SDS in all patients available for analysis. Among those 35 patients with contralateral anomalies (mainly obstructive changes and vesico-ureteral reflux), all 3 patients with contralateral changes suggestive of hypoplasia and 22% with obstruction, but only 1/7 (14%) with reflux showed elevated serum creatinine level >2 SDS. CONCLUSION: Renal function seems to be slightly impaired in patients with unilateral multicystic renal dysplasia. The apparent tendency to regression of the dysplastic kidney and no difference in the number of complications justify a conservative management rather than operative intervention. PMID- 9727852 TI - Early vitamin K deficiency bleeding after maternal phenobarbital intake: management of massive intracranial haemorrhage by minimal surgical intervention. AB - Vitamin K deficiency bleeding within the first 24 h of life is caused in most cases by maternal drug intake (e.g. coumarins, anticonvulsants, tuberculostatics) during pregnancy. Haemorrhage is often life-threatening and usually not prevented by vitamin K prophylaxis at birth. We report a case of severe intracranial bleeding at birth secondary to phenobarbital-induced vitamin K deficiency and traumatic delivery. Burr hole trepanations of the skull were performed and the subdural haematoma was evacuated. Despite the severe prognosis, the infant showed an unexpected good recovery. At the age of 3 years, neurological examinations were normal as was the EEG at the age of 9 months. CT showed close to normal intracranial structures. CONCLUSION: This case report stresses the importance of antenatal vitamin K prophylaxis and the consideration of a primary Caesarean section in maternal vitamin K deficiency states and demonstrates the successful management of massive subdural haemorrhage by a limited surgical approach. PMID- 9727854 TI - Multicystic kidney dysplasia: a prospective study on the natural history of the affected and the contralateral kidney. AB - In a 6-year period, 41 young infants with multicystic kidney dysplasia were seen in our department. In 30 cases, the diagnosis had already been suspected by prenatal ultrasonography. A prospective protocol was proposed to the parents which comprised ultrasound evaluation every 3 months until the age of 24 months and renal function assessment at the age of 18 months. In 33 patients, the study was completed as scheduled. At the start of the study, the maximal diameter of the multicystic kidney was above the mean length of normal kidneys in all cases where precise measurement was possible. Within 24 months, 7 of the dysplastic kidneys disappeared, 20 regressed in size, 1 remained unchanged and only 5 increased in size. Between the age of 0 to 3 months, renal length of the contralateral kidney was within the normal range in 19 infants and above +2SD in 14 cases. At the age of 18 to 24 months, renal length was, with few exceptions, between 0 and +4SD. Inulin clearance was normal in all 33 individuals with a median value of 112 ml/min per 1.73 m2. CONCLUSION: As a rule, multicystic kidneys shrink in the first 2 years of life. In most cases the contralateral kidney maintains a normal renal function as a consequence of progressive compensatory hypertrophy. PMID- 9727855 TI - Childhood immunisation in the European Union. Confederation of European Specialists in Paediatrics (CESP). AB - Harmonisation of working conditions is one of the major aims of the European Union. The Confederation of European Specialists in Paediatrics has elaborated documents on the harmonisation of immunisations in 1987 and 1992. The new and completely updated version focuses on the variations of immunisation practices and schedules and gives criteria for minimal agreement of immunisation schedules among the member countries. Catch-up vaccinations and false contra-indications are especially stressed. PMID- 9727856 TI - Risk and preventive factors for cot death in The Netherlands, a low-incidence country. AB - In the Netherlands an 18 months case control study into cot death was undertaken as part of the European Concerted Action (ECAS) on sudden infant death syndrome to determine the relative risk of prone sleeping and other sleep practices. Physicians in the Netherlands were asked to report to the study centre all sudden and unexpected deaths of children between 1 week and 2 years of age. Non cot death cases were deleted from further analysis after a consensus was reached by three pathologists, not primarily involved in the post mortem diagnosis. A positive response of families was achieved in 91% of cases registered in the Central Bureau of Statistics. The study comprised 73 cot deaths and 146 controls, two for each case and matched for date of birth. All families were visited at home for completion of a questionnaire. The cot death rate has dropped considerably over the past 10 years after the recommendations on supine sleeping to a low of 0.26 per 1000 live born infants. In addition to the ECAS objective, we wanted to establish whether previously found risk factors are still valid in the present situation or that new factors might have emerged, some of them possibly protective. CONCLUSION: Placing an infant prone or on side on last occasion, secondary prone position (not placed prone but turned to prone), inexperienced prone sleeping and use of a duvet, leading to head and body being covered, were shown to be risk factors. Preventive factors were using a cotton sleeping-sack and a dummy. Even in a low incidence country, such as the Netherlands, there are indications that further prevention is possible. PMID- 9727857 TI - Sudden pneumoscrotum in a ventilated infant. Sudden pneumoscrotum as clinical sign of a tension pneumothorax in an infant with tracheal stenosis. PMID- 9727858 TI - Lymphocytic hypophysitis and central diabetes insipidus during adolescence: what are the criteria for diagnosis? PMID- 9727859 TI - Antroduodenal motor function and gastro-oesophageal reflux in neurologically impaired children, adolescents and young adults. PMID- 9727860 TI - Umbilical venous catheterization and development of Banti syndrome: the possible role of the factor V Leiden mutation. PMID- 9727861 TI - Effect of nitric oxide inhalation on pulmonary hypertensive crisis in a case of aortic origin of the right pulmonary artery. PMID- 9727862 TI - Thrombocytopenic purpura secondary to brucellosis. PMID- 9727863 TI - Influence of preformed alpha-helix and alpha-helix induction on the activity of cationic antimicrobial peptides. AB - One prominent class of cationic antibacterial peptides comprises the alpha helical class, which is unstructured in free solution but folds into an amphipathic alpha-helix upon insertion into the membranes of target cells. To investigate the importance of alpha-helicity and its induction on interaction with membranes, a series of peptides was constructed based on a hybrid of moth cecropin (amino acids 1-8) and bee melittin (amino acids 1-18) peptides. The new peptides were predicted to have a high tendency to form alpha-helices or to have preformed alpha-helices by virtue of construction of a lactam bridge between glutamate and lysine side-chains at positions i and i + 4 at various locations along the primary sequence. In two examples where the use of lactam bridge constraints induced and stabilized alpha-helical structure in benign (aqueous buffer) and/or hydrophobic medium, there was a decrease in antibacterial activity relative to the linear counterparts. Thus the preformation of alpha-helix in solution was not necessarily beneficial to antimicrobial activity. In the one case where the lactam bridge did result in increased antibacterial activity (lower minimal inhibitory concentration values) it did not increase alpha-helical content in benign or hydrophobic medium. Broadly speaking, good activity of the peptides against Pseudomonas aeruginosa correlated best (r2 = 0.88) with a helican parameter which was calculated as the induction of alpha-helix in a membrane-mimicking environment divided by the alpha-helix formation under benign conditions. Interestingly, the activity of the lactam bridge peptide constructs correlated in part with alterations in bacterial outer or cytoplasmic membrane permeability. PMID- 9727864 TI - Design of peptides with alpha,beta-dehydro residues: synthesis, crystal structure and molecular conformation of N-Boc-L-Ile-deltaPhe-L-Trp-OCH3. AB - The dehydro-peptide Boc-L-Ile-deltaPhe-L-Trp-OCH3 was synthesized by the azlactone method in the solution phase. The peptide was crystallized from methanol in an orthorhombic space group P2(1)2(1)2(1)with a = 10.777(2), b = 11.224(2), c = 26.627(10) A. The structure was determined by direct methods and refined to an R value of 0.069 for 3093 observed reflections [I > or = 2delta(I)]. The peptide failed to adopt a folded conformation with backbone torsion angles: phi1 = 90.8(8)degrees, psi1 = -151.6(6)degrees, phi2 = 89.0(8)degrees, psi2 = 15.9(9)degrees, phi3 = 165.7(7)degrees, psi3T = 166.0(7)degrees . A general rule derived from earlier studies indicates that a three-peptide unit sequence with a deltaPhe at the (i + 2) position adopts a beta turn II conformation. Because the branched beta-carbon residues such as valine and isoleucine have strong conformational preferences, they combine with the deltaPhe residue differently to generate a unique set of conformations in such peptides. The presence of beta-branched residues simultaneously at both (i + 1) and (i + 3) positions induces unfolded conformations in tetrapeptides, but a beta branched residue substituted only at (i + 3) position can not prevent the formation of a folded beta-turn II conformation. On the other hand, the present structure shows that a beta-branched residue substituted at the (i + 1) position prevents the formation of a beta-turn II conformation. These observations indicate that a beta-branched residue at the (i + 1) position prevents a folded conformation whereas it cannot generate the same degree of effect from the (i + 3) position. This may be because of the trans disposition of the planar deltaPhe side-chain with respect to the C=O group in the residue. The molecules are packed in an anti-parallel manner to generate N2-H2...O2 (-x, y -1/2, -z + 3/2) and N3epsilon1-H3epsilon1 ...O1(-X, y -1/2, -z + 3/2) hydrogen bonds. PMID- 9727866 TI - Incomplete trifluoroacetic acid deprotection of asparagine-trityl-protecting group in the vicinity of a reduced peptide bond. AB - During the Fmoc solid-phase synthesis of reduced peptide bond analogues, we observed that the trityl protection of an asparagine residue in the vicinity of a reduced peptide bond is not cleaved completely after the final trifluoroacetic acid deprotection step. The relative position of the Asn side-chain amine and of the aminomethylene bond as well as the preferential protonation of the secondary amine can be used to explain this phenomenon. We show that longer deprotection times or the use of methyl-trityl protection partially improves the yield of the Asn-deprotected peptide whereas xanthenyl protection totally overcomes this problem. PMID- 9727865 TI - Structure-function analysis of the Saccharomyces cerevisiae tridecapeptide pheromone using alanine-scanned analogs. AB - Twenty-six peptide analogs of the Saccharomyces cerevisiae alpha-factor, a tridecapeptide mating pheromone (W1H2W3L4Q5L6K7p8G9Ql0P11M12Y13) with either L- or D-alanine replacement of each amino acid residue (Ala-scanned) and with the isosteric replacement of methionine at position 12 by norleucine, were synthesized, purified to homogeneity and assayed for biological activity and receptor binding. Two new and effective antagonists, [D-Ala3,Nle12]alpha-factor and [D-Ala4,Nle12]alpha-factor, were found among the series, and the [D Ala10,Nle12]alpha-factor demonstrated a marked ability to increase the biological activity of [Nle12]alpha-factor without having any effect by itself. One analog, the [L-Ala1 alpha-factor, showed a 3-fold increase in bioactivity over the [Nle12]alpha-factor, although its binding to the alpha-factor receptor was about 70-fold less than [Nle12]alpha-factor. Residues near the carboxyl terminus contributed more strongly to receptor binding than other residues, whereas those near the amine terminus of the alpha-factor played an important role in signal transduction. The effect of insertion of D-Ala residues at positions 7, 8, 9 and 10 on bioactivity and receptor binding of the peptide suggested a specific positioning role of the central loop in establishing optimal contacts between the receptor and the ends of the pheromone. We conclude that the alpha-factor may be divided into segments with dominant roles in forming the biologically active pheromone conformation, in receptor binding and in initiating signal transduction. The discovery of such relationships was made possible by the systematic variation of each residue in the peptide and by the testing of each analog in highly defined biological and binding assays. PMID- 9727867 TI - Role of conformational constraints of position 7 of the disulphide bridge of h alpha-CGRP derivatives in their agonist versus antagonist properties. AB - Previous structure-activity studies have shown that the disulphide bridge of calcitonin gene-related peptide (CGRP) is important for the highly potent, CGRP receptor-mediated effects of this peptide. In this study penicillamine (Pen) was substituted for one or both of the cysteinyl residues to determine conformational and topographical properties of the disulphide bridge favourable for binding to CGRP receptors and/or receptor activation. Pen constrains the conformational flexibility of disulphide bridges in other peptides. Binding affinities were measured using a radioligand binding assay with membranes prepared from pig coronary arteries and 125I-h-alpha-CGRP. Functional effects were characterized using a previously reported pig coronary artery relaxation bioassay. The binding affinity of [Pen2]h-alpha-CGRP was not significantly different from that of h alpha-CGRP. All other analogues showed reduced affinity for CGRP receptors. [Pen2]h-alpha-CGRP also caused relaxation of coronary arteries. The remaining analogues either caused relaxation with significantly reduced potency or failed to relax the arteries at concentrations up to 1 x 10(-5)M. All analogues that did not relax coronary arteries contained a D-Pen in position 7 and inhibited CGRP induced relaxation. [D-Pen2,7]h-alpha-CGRP was the most potent antagonist with a K8 value of 630 nM. This affinity is similar to that of the classical CGRP receptor antagonist, h-alpha-CGRP(8-37), on these arteries (KBs 212 nM). These studies show that modifying the topography of the disulphide bridge can cause large and variable effects on ligand binding and activation of CGRP receptors. The contribution of position 7 to the conformation and topography of the disulphide bridge of h-alpha-CGRP is crucial to the future design of agonists of CGRP receptors. Furthermore, position 7 is important for the development of new CGRP receptor antagonists with structures based on the whole sequence of h-alpha CGRP. PMID- 9727868 TI - Solid-phase synthesis and on-resin cyclization of a disulfide bond peptide and lactam analogues corresponding to the major antigenic site of HIV gp41 protein. AB - A cyclic peptide that spans the major antigenic determinant of the human immunodeficiency virus (HIV) glycoprotein 41 (gp41) has been synthesized according to various strategies. For immunodiagnostic applications, biotin was added at the N-terminus of the peptide and aminohexanoic acid was used as a spacer. Polymer-supported oxidations were carried out in a variety of ways with thallium (III) trifluoroacetate. The biotinylcyclic peptide was released from the support using trimethylsilyl trifluoromethane sulfonate and various scavengers. The efficacy of these different cyclization and cleavage procedures was compared. Side reactions were studied, and a simple and efficient procedure was set up to monitor peptide cyclization by mass spectrometry. In a second series of syntheses the disulfide bridge was replaced by an amide bond. For this purpose, an aspartic acid derivative and a diaminopropionic acid were introduced during the synthesis in place of the two cysteine residues in the parent sequence. On-resin cyclization was performed and led to a major side-product identified as a piperidide. This undesired base-mediated side reaction was prevented when, instead of piperidine, 1,8-diazabicyclo-[5.4.0]undec-7-ene was used for fluorenylmethyl ester deprotection. Reactivity of these peptides with different patients' sera and with a monoclonal antibody directed against the whole gp41 was tested using an enzyme-linked immunosorbent assay. PMID- 9727869 TI - Identification of critical residues of staphylococcal enterotoxin B for lymphomonocyte proliferation and cytokine production. AB - Superantigens bind to major histocompatibility complex class II molecules and stimulate large numbers of T cells expressing particular Vbeta elements of the T cell receptor. Staphylococcal enterotoxin B (SEB) is a bacterial superantigen that causes food poisoning and toxic-shock syndrome. The toxicity of SEB is thought to be mediated by T-cell stimulation and cytokine production. Different regions of the SEB molecule are important for mitogenic activity. To identify critical residues of SEB in the region 124-1 54, which competitively inhibits the mitogenic activity of the toxin, we used the synthetic peptide approach and alanine scanning mutagenesis as a probe. We synthesized eight peptides with alanine replacement of all residues in the SEB sequence 131-138 and tested them for the capacity to inhibit both SEB-induced proliferation of human lymphomonocytes and the production of tumor necrosis factor alpha and interferon gamma. Mutation to alanine of the residue Thr 133 improved the inhibition of SEB induced proliferation and cytokine production, whereas the substitution of Ser 131 also increased the inhibition, albeit to a lesser degree. The peptide obtained by substitution of Val 136 with alanine was unable to inhibit SEB induced proliferation and cytokine production, suggesting that Val 136 is essential for mitogenic activity. Thus hydrophobic interactions apparently are very important for mitogenic activity. The identification of critical residues in this active site in the SEB and the computer modeling based on crystal X-ray data contribute to a better understanding of the molecular mechanism of the superantigen and may be useful for therapeutical applications. PMID- 9727870 TI - Isolation and characterization of a group of oligopeptides related to oxidized glutathione from the root of Panax ginseng. AB - Six gamma-glutamyl oligopeptides were isolated for the first time from aqueous methanol extracts of Panax ginseng root by using column chromatography on ion exchange resin, gel filtration and reverse-phase high-performance liquid chromatography. Their structures had been established with the methods of amino acid analysis, N-terminal, C-terminal determination and double-coupling sequence analysis. They were: P-I (N-gamma-glutamylcystinyl-bis-glycine), P-ll (gamma glutamylcysteinylglycine disulfide, oxidized glutathione), P-III (N,N'-bis-gamma glutamylcystinylglycine), P-IV (gamma-glutamylcysteinylglycinamide disulfide), P V (N-gamma-glutamylglycylcysteine disulfide), P-VI(gammaglutamylarginine); five of them are related to oxidized glutathione. The structures were further confirmed by the chemical synthesis. As far as we know, P-V (N-gamma glutamylglycylcysteine disulfide) is a new biologically active peptide which exhibits somnogenic effect and is more potent than that of P-II. PMID- 9727872 TI - Combined solid-phase/solution synthesis of a 31-residue vasoactive intestinal peptide analog: general method for repetitive coupling of fragments without isolation and purification of intermediates. AB - A novel analog of vasoactive intestinal peptide (VIP) has been reported which exhibits high potency and enhanced duration of in vivo biological activity. This VIP analog, cyclo-(Lys21-Asp25)Ac[Glu8 Lys12 Nle17 Ala19, Asp25 Leu26,Lys27,28,Gly29,30,Thr31]-VIP, which also has a lactam bridge, has been reported to have relaxant effects that are significantly more potent than other beta-agonists such as salbutamol and salmeterol. Because it has potential use for the treatment of bronchial asthma in humans, various convergent syntheses were evaluated to enable the economic preparation of large quantities of this medium sized hentriacontapeptide. From these studies we developed a combined solid phase/solution synthesis which uses four protected fragments (each prepared by solid-phase synthesis with highly acid-labile resins) possessing Nalpha-9 fluorenylmethyloxycarbonyl and side-chain tert-butyl protection. Only equivalent amounts of each fragment were required to achieve near-quantitative coupling reactions using N-[(1H-benzotriazol-1-yl)(dimethylamino)methylene]-N-methylmeth anaminium hexafluorophosphate N-oxide/N-hydroxybenzotriazole. All reagents and side products were removed at each stage by simple extraction procedures. Final deprotection was carried out with 90% trifluoroacetic acid. Under these conditions only low levels of epimerization were observed (<2%). These diastereoisomers and other trace impurities were removed from the product in a single purification by preparative high-performance liquid chromatography. The procedure has been scaled up (10-g scale) and the final product obtained in an overall (nonoptimized) yield of 24%. This procedure for the repetitive coupling of fragments, without isolation of intermediates, may be generally applicable for the economic synthesis of other medium-sized and longer peptides. PMID- 9727871 TI - Contribution to the synthesis of aureobasidin A. Synthesis of cyclopeptolides containing the sequence leucyl-N-methyl-beta-hydroxyvalyl-(2R)-oxy-(3R)-methyl pentanoi c acid. AB - The efficient antimycotic agent aureobasidin A, isolated from the culture broth of Aureobasidium pullulans R 106, and the [(R)-Pro9]-aureobasidin A were prepared starting from benzyl N-Boc-N-methyl-(S)-beta-triethylsiloxyvalinate, the synthesis of which is described here. The easy accessibility of the tripeptolide benzyl Boc-leucyl-N-methyl-beta-hydroxyvalyl-(2R)-oxy-(3R)-methylpentanoa te [Boc Leu-HOMeVal-(R)-HMP-OBn] facilitates the construction of the cyclopeptolides 28, 34, 45 and 47. The peptide bonds of the N-methylamino acids were formed with the help of O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophophate. The rings of [(R)-Pro9]-aureobasidin A and of cyclopeptolides 28, 34, 45 and 47 were closed by way of pentafluorophenyl esters. The ring of aureobasidin A could only be formed with bromo-tris-pyrrolidinophosphonium hexafluorophosphate. PMID- 9727873 TI - Sensitization of cells to ionizing radiation by chlorin e6Na. AB - We have investigated whether a hydrophilic photosensitizer, chlorin e6Na (Ce6Na), can sensitize cells to ionizing radiation. When V79-1 cells were pretreated with Ce6Na for 2 hr before receiving a dose of 2-14 Gy of irradiation, the cells became sensitive to X-irradiation. The sensitizing effect of Ce6Na depended on the dose of Ce6Na. The sensitizing effect also depended on the length of the treatment period before X-irradiation up to 4 hr, but not on the length of a treatment period after X-irradiation. Intracellular concentrations of Ce6Na were increased linearly after incubation with Ce6Na for periods of up to 4 h. The dose modifying factor calculated from the survival curve was 1.25. PMID- 9727874 TI - Time-dependent changes in CT-based dosimetry of I-125 prostate brachytherapy. AB - To determine the effect of time between prostate brachytherapy and evaluation CT scan on calculated target coverage. CT scans from 11 consecutive, unselected patients with stage T1 or T2 prostatic carcinoma who had transperineal I-125 implants at MSKCC in 1996 were analyzed for target coverage at 0, 2, and 6 months after implantation. The outer margins of the prostate were outlined on each CT section by a single investigator. In each CT plane, the prescription isodose (150 Gy) was overlaid on the target contour from the postimplant CT to calculate the integral Dose-Volume Histogram. The postimplant target volume on the day of the implant ranged from 93% to 160% of the preimplant volume (average: 117%). In all patients, the target size returned to the preimplant size or smaller within 2 months of the procedure and was relatively stable between 2 to 6 months. Immediately following the implant, an average of 84% of the target (range: 73 98%) was covered by the 150 Gy isodose line. Consistent with changes in the target volume over time, the target coverage increased from an average of 84% to 90% between 0 to 2 months and did not change substantially between 2 and 6 months. There was minimal source loss from the target area after the implant. It was concluded that temporary, postimplant swelling will increase the target volume, making target coverage inferior to what would be calculated if a dosimetry scan was taken sometime later, after the acute swelling has subsided. Until the clinical significance of the effect of postimplant volume changes is better defined, we are continuing to obtain evaluation scans on the day of the implant. PMID- 9727875 TI - Treatment of localized non-Hodgkin's lymphomas of the head and neck: focusing on cases of non-lethal midline granuloma. AB - This report clarifies the prognostic factors for survival in localized non Hodgkin's lymphoma (NHL) of the head and neck and defines optimal regimens for this disease. One hundred-seven untreated patients with Stage I or II NHL of the head and neck were treated with involved field radiation therapy for orbital, nasal, or paranasal lymphoma and extended field radiation for Waldeyer's ring or neck lymphoma. Radiation doses were 39-48 Gy. In the latter half of the study, adjuvant chemotherapy was administered. Of 107 patients, 95 achieved complete response (CR). Of the 12 patients that did not achieve CR, 9 had nasal T-cell lymphoma (NTL) of the lethal midline granuloma type (LMG-NTL). Only one patient who obtained CR relapsed in a previously irradiated area. Age, sex, stage, bulky mass, number of involved sites, LMG-NTL, histologic subtypes, radiation dose, and adriamycin dose were analyzed for prognostic significance for disease-specific survival in NHL by multivariate analysis. LMG-NTL was the most significant prognostic factor (P < 0.001). Patients with higher age also experienced a higher relative risk than patients of > or =60 years of age (P = 0.0063). Dose of adriamycin reached the borderline significance (P = 0.0600). Radiotherapy is excellent for obtaining local control of head and neck NHL. Randomized trials are required to determine the appropriate radiation field and dose in patients previously treated with chemotherapy. LMG-NTL and age were the significant prognostic factors for disease-specific survival. PMID- 9727876 TI - Pathologic features from prostate needle biopsy and prognosis after I-125 brachytherapy. AB - To evaluate the role of detailed pathologic features in predicting outcome for early-stage prostate cancer treated with I-125 brachytherapy. The pretreatment biopsy slides of 103 patients with T1/T2 and Gleason scores of 4-7 prostatic carcinoma, which was treated by transperineal I-125 implantation, were reviewed retrospectively by a single pathologist (P.B.G.). Biochemical tumor control rates [prostate-specific antigen (PSA) below 1.0] were correlated with pretreatment PSA, Gleason score, the amount of tumor in the biopsy samples, and the presence of perineural invasion. In Cox proportional-hazard, multivariate analysis, the strongest predictors of failure were pretreatment PSA above 10 ng/ml (P = 0.013) and the length of the biopsy specimen replaced by tumor (P = 0.15). The percent of biopsy tissue replaced by tumor (P = 0. 74), perineural invasion (P = 0.78), and Gleason score (P = 0.66) were less predictive of prognosis. It was concluded that pretreatment PSA is the strongest predictor of biochemical failure. Detailed assessment of pathological features on needle biopsy added little prognostic information beyond that of pretreatment PSA alone. Like all other prognostic parameters for prostate cancer, there is considerable overlap in pathologic features between those patients who will or will not be controlled biochemically. PMID- 9727877 TI - Accelerated radiotherapy regimen for malignant gliomas using stereotactic concomitant boosts for dose escalation. AB - The purpose of this pilot study was to determine the feasibility and toxicities of an accelerated treatment program by using a concomitant stereotactic radiotherapy boost given weekly during a course of standard external-beam irradiation (EBXRT) in patients with malignant gliomas. Twelve patients underwent biopsy or subtotal resection of a malignant glioma and were enrolled on the protocol, which delivered 44 Gy-EBXRT and a 12-Gy stereotactic radiotherapy boost given on 3 consecutive weeks of treatment for a total dose of 80 Gy over 33 days. Three patients with anaplastic astrocytoma and nine patients with glioblastoma multiforme had median survival times of 33 months and 16 months, respectively. All of the tumor recurrences were within or were closely adjacent to the region of high-dose irradiation. None of the patients required a treatment break, and there were no acute complications. Two patients developed seizures in the follow up period, and four patients were diagnosed with radionecrosis at the time of the second operation. The treatment program was found to be feasible and was well tolerated, and it resulted in a rate of late complications similar to those of radiosurgery or interstitial brachytherapy. PMID- 9727879 TI - Isoseparation curves: a mechanism for optimizing off-axis dose homogeneity of intact breast irradiation. AB - The purpose of this paper is to determine whether using off-axis isoseparation curves to optimize the collimator rotation angle improves dose homogeneity. Eleven intact breast irradiation patients underwent computerized tomography (CT) treatment planning with 1 cm abutting slices. Central plane treatment planning, using 6 MV photons, tissue inhomogeneity corrections, and isocentric opposed tangent treatment fields, was performed. Collimators were rotated to match chest wall slope through the use of a beam's-eye-view setting. Patient separations were measured from the apex of the breast to the posterior field border on each axial CT slice. Sagittal-plane isoseparation curves were constructed from these measurements. Using these curves, the collimator rotation that minimized off-axis separation differences was determined. A comparison of off-axis dose inhomogeneity was performed for patients with a > or =10 degrees difference between this optimized collimator angle and the angle determined by chest wall slope. These comparative treatment plans differed only with respect to collimator angle rotation. The mean optimal collimator rotation angle differed significantly from the mean rotation angle which matched the chest wall slope (5.4 degrees vs. 11 degrees, respectively, P < 0.001). Four of the 11 patients had rotation angle differences of 10 degrees. In these patients, the optimization of collimator angle reduced the percentage of breast volume to "that" received > or =110% of the prescribed dose. For the patient with the largest breast size to the patient with the smallest breast size the decreases were, respectively, 5% (15% to 10%), 3% (24% to 21%), 1% (4% to 3%), and 1% (0.9% to 0%). The mean reduction in dose inhomogeneity was greatest in the inferior breast quadrants. At 6 cm and 4 cm off axis, the mean reductions in the percentages of the breast tissue to "that" received 110% of the prescribed dose were respectively 15.1% and 5.3 %. Optimizing the collimator angle through the use of isoseparation curves decreases dose inhomogeneity. The greatest improvements are in the inferior quadrants of the intact breast. The improved dose homogeneity may have clinical relevance in the treatment of patients with large breast sizes. PMID- 9727878 TI - Influence of timing on the dosimetric analysis of transperineal ultrasound guided, prostatic conformal brachytherapy. AB - Postoperative computed tomography (CT)-based dosimetric analysis of transperineal ultrasound-guided conformal prostate brachytherapy provides detailed information regarding the coverage and uniformity of the implant. However, there is no generally accepted standard for the optimal timing of the postoperative dosimetry. This report details dosimetric analysis and the effect of timing based upon CT and orthogonal film evaluation for ten unselected patients implanted with either iodine-125 (125I) or palladium-103 (103Pd). Within 2 hours after implantation, patients underwent a CT scan and the first of four sequential sets of orthogonal films. Subsequent orthogonal films were obtained on days 3, 14, and 28 postimplant. CT-based dosimetry revealed coverage of the prostate to the prescribed minimal peripheral dose (mPD) at 93.1 +/- 3.6% of the volume, the prostate volume receiving 150% of mPD was 38.2 +/- 8.7%, and the urethral and rectal doses were 114 +/- 12% and 78 +/- 19% of mPD, respectively. The implanted seeds seen on orthogonal films acted as markers for temporal changes in prostate dimensions, and the standard deviation of each dimension was used as input in an ellipsoidal volume calculation. Seed coordinates were self normalized to the center of gravity of each two-dimensional view and were measured relative to the linear regression line in the superior-inferior direction. The reproducibility of the anteroposterior (AP) film setup in terms of temporal variation in the angle of the regression line was markedly better than that of the lateral films, 1.8 degrees +/- 1.2 degrees vs. 4.3 degrees +/- 2.6 degrees, respectively. Dimensional contraction from day 0 to day 28 averaged 11.3% in the superior inferior direction, 8.5% in the AP/PA (posteroanterior) direction, and 2.5% in the right-left lateral direction. This translated into a volume change of 20.9% (ranged 11.6-31.6%), which was determined by using the ellipsoid method. The half life for edema resolution was 10.6 +/- 1.8 days (range 8.6-14.3 days). However, because of variability in the degree and extent of edema and its rate of resolution, we believe that it may be futile to define a single point in time as the most accurate indicator of the postoperative dose distribution. Rather, it may be preferable to accept universal standardization of timing and methodology for CT-based postoperative dosimetry, which would facilitate comparison of results between centers and maximize the information content of that single measurement. We conclude that day 0 represents the optimal time, because dosimetric evaluation at that time minimizes patient discomfort and inconvenience (a catheter is already in place), provides information about edema when it is near its maximum extent, and provides prompt closure of the learning loop and, as such, hopefully will result in improved implantation techniques and results. PMID- 9727880 TI - How do we know? Reflections on qualitative research in diabetes. PMID- 9727881 TI - Learning to use troglitazone. PMID- 9727882 TI - Attitudes of primary care providers toward diabetes: barriers to guideline implementation. AB - OBJECTIVE: Primary care providers have been slow to adopt standards of care for diabetes, and continuing medical education (CME) programs have been minimally effective in changing provider behavior. The objective of this study was to explore the previously reported finding that attitudes, rather than knowledge, may impede primary care provider adherence to standards of care. RESEARCH DESIGN AND METHODS: Study participants included 31 primary care providers attending an eight-session CME program on diabetes. Providers rated on a 10-point scale how the treatment of diabetes compared with that of five other chronic conditions (hypertension, hyperlipidemia, angina, arthritis, and heart failure; 1 = easier to 10 = harder; midpoint 5.5). In a subsequent open-ended qualitative interview, providers explained their scale ratings. RESULTS: Diabetes was rated as significantly harder to treat than hypertension (24 of 30 >5.5; P < 0.001) and angina (20 of 30 >5.5; P = 0.03). A majority also rated hyperlipidemia (18 of 30) and arthritis (18 of 30) as easier to treat than diabetes. Explanatory themes underlying provider frustrations with diabetes include characteristics of the disease itself and the complexity of its management, and a perceived lack of support from society and the health care system for their efforts to control diabetes. CONCLUSIONS: CME that addresses provider attitudes toward diabetes in addition to updating knowledge may be more effective than traditional CME in promoting adherence to standards of care. Additional changes are needed in our health care system to shift from an acute to a chronic disease model to effectively support diabetes care efforts. PMID- 9727883 TI - Accuracy of calculated serum low-density lipoprotein cholesterol for the assessment of coronary heart disease risk in NIDDM patients. AB - OBJECTIVE: To evaluate the accuracy of LDL cholesterol calculated with Friedewald's equation in the assessment of cardiovascular risk in NIDDM patients. RESEARCH DESIGN AND METHODS: The calculation of LDL cholesterol according to Friedewald's formula was compared with the measurement of LDL cholesterol separated by ultracentrifugation in 151 NIDDM patients with fairly good metabolic control (HbA1c < or =10%) and in 405 nondiabetic subjects. RESULTS: Measured and calculated LDL cholesterol was found to be well correlated in both diabetic (r = 0.95) and nondiabetic (r = 0.97) subjects. Compared with measured LDL cholesterol, the calculated LDL cholesterol differed by > or =10% in 34% of samples from diabetic patients and in 26% of samples from nondiabetic subjects (chi(2) = 3.885, P < 0.05). The percentage of error increased when the serum triglyceride (TG) level was > or =200 mg/dl (2.26 mmol/l) and when the ratio of VLDL cholesterol to TG was <0.20 or >0.29 in both groups of subjects. Although the percentage of error from calculated LDL cholesterol was greater in diabetic than in nondiabetic subjects because of the greater prevalence of hypertriglyceridemia in the former group, the misclassification of coronary heart disease risk, according to the cutoff points of the National Cholesterol Education Program (NCEP), was similar in the two groups (25% in diabetic and 22% in nondiabetic subjects). In both groups of patients, the misclassification of coronary heart disease risk was higher when calculation of LDL cholesterol produced values near the cutoff points. CONCLUSIONS: Although accuracy in the estimation of LDL cholesterol is less than ideal, Friedewald's equation seems to be of value in the correct assignment of coronary heart disease risk classes in the great majority of diabetic as well as nondiabetic subjects. Caution must be exercised for subjects in whom calculated LDL cholesterol is close to the cutoff points of the NCEP guidelines. PMID- 9727884 TI - The third version of the Diabetes Attitude Scale. AB - OBJECTIVE: The objective of this study was to develop a third version of the Diabetes Attitude Scale (DAS-3) that is congruent with current scientific knowledge about diabetes, has improved subscale internal reliability scores, and is shorter than the earlier versions of this instrument. RESEARCH DESIGN AND METHODS: The second DAS was revised and rewritten by a panel of diabetes experts, including patients, associated with the University of Michigan Diabetes Research and Training Center. The revised version of the instrument was sent to physicians, nurses, dietitians, and patients with diabetes. Completed and usable questionnaires were obtained from 384 patients with diabetes, 321 physicians, 540 nurses, and 569 dietitians. The total number of surveys used for these analyses was 1,814. RESULTS: The study resulted in a revised DAS with 33 items and five discrete subscales. The subscales were attitudes toward the following: 1) need for special training to provide diabetes care, 2) seriousness of type 2 diabetes, 3) value of tight glucose control, 4) pyschosocial impact of diabetes, and 5) attitude toward patient autonomy. Overall, the subscale reliabilities of the DAS 3 were superior to the earlier versions of the scale. CONCLUSIONS: The DAS-3 is a valid and reliable general measure of diabetes-related attitudes and is most suitable for comparisons across different groups of health care professionals and/or patients. The DAS-3 is also suitable for the evaluation of patient and/or professional education programs if those programs focus on the specific topic areas measured by the five DAS-3 subscales. PMID- 9727885 TI - Effects of changing diagnostic criteria on the risk of developing diabetes. AB - OBJECTIVE: The American Diabetes Association (ADA) has recommended that the fasting plasma glucose (FPG) level used to diagnose diabetes be changed from 7.8 mmol/l (the level recommended by the National Diabetes Data Group [NDDG] in 1979) to 7.0 mmol/l. We examined the impact of this change on rates of progression to overt diabetes from different levels of FPG. RESEARCH DESIGN AND METHODS: Using the laboratory database of Mayo Clinic, we assembled a cohort of 8,098 nondiabetic Olmsted County residents 40 years of age or older on 1 July 1983. Subjects were followed for a median of 9 years. RESULTS: Among 7,567 individuals with follow-up FPG data, 778 (10.3%) progressed to ADA diabetes and 513 (6.8%; P < 0.0001) progressed to NDDG diabetes. The risk of developing ADA diabetes was 7, 19, and 39% for individuals with initial FPG values in the ranges of <5.6, 5.6 6.0, and 6.1-6.9 mmol/l, respectively. For progression to NDDG diabetes, the respective risks were 3, 11, and 25%. A clear gradient of risk was observed within the "normal" range of FPG (<5.6 mmol/l). Among the 793 individuals who developed ADA diabetes, 222 (29%) developed NDDG diabetes simultaneously and 291 (37%) developed NDDG diabetes later. In all FPG subgroups, progression to ADA diabetes occurred approximately 7 years sooner than progression to NDDG diabetes. CONCLUSIONS: The baseline level of FPG is a major predictor of an individual's risk of developing diabetes. The proposed change in the diagnostic criteria for diabetes will lead to earlier diagnosis among individuals who are destined to develop the disease. PMID- 9727886 TI - Global burden of diabetes, 1995-2025: prevalence, numerical estimates, and projections. AB - OBJECTIVE: To estimate the prevalence of diabetes and the number of people with diabetes who are > or =20 years of age in all countries of the world for three points in time, i.e., the years 1995, 2000, and 2025, and to calculate additional parameters, such as sex ratio, urban-rural ratio, and the age structure of the diabetic population. RESEARCH DESIGN AND METHODS: Age-specific diabetes prevalence estimates were applied to United Nations population estimates and projections for the number of adults aged > or =20 years in all countries of the world. For developing countries, urban and rural populations were considered separately RESULTS: Prevalence of diabetes in adults worldwide was estimated to be 4.0% in 1995 and to rise to 5.4% by the year 2025. It is higher in developed than in developing countries. The number of adults with diabetes in the world will rise from 135 million in 1995 to 300 million in the year 2025. The major part of this numerical increase will occur in developing countries. There will be a 42% increase, from 51 to 72 million, in the developed countries and a 170% increase, from 84 to 228 million, in the developing countries. Thus, by the year 2025, >75% of people with diabetes will reside in developing countries, as compared with 62% in 1995. The countries with the largest number of people with diabetes are, and will be in the year 2025, India, China, and the U.S. In developing countries, the majority of people with diabetes are in the age range of 45-64 years. In the developed countries, the majority of people with diabetes are aged > or =65 years. This pattern will be accentuated by the year 2025. There are more women than men with diabetes, especially in developed countries. In the future, diabetes will be increasingly concentrated in urban areas. CONCLUSIONS: This report supports earlier predictions of the epidemic nature of diabetes in the world during the first quarter of the 21st century. It also provides a provisional picture of the characteristics of the epidemic. Worldwide surveillance of diabetes is a necessary first step toward its prevention and control, which is now recognized as an urgent priority. PMID- 9727887 TI - Population-based assessment of the level of care among adults with diabetes in the U.S. AB - OBJECTIVE: To estimate the levels of use of preventive care and to identify correlates of such care among people with diabetes in the U.S. RESEARCH DESIGN AND METHODS: A cross-sectional study was conducted using a sample of 2,118 adults, age > or =18 years, with self-reported diabetes in 22 states that participated in the 1994 Behavioral Risk Factor Surveillance System. Most subjects were age > or =45 years (83%), women (51%), and white (75%) and were diagnosed at ages > or =30 years (83%), had type 2 diabetes (89%), and were not using insulin (66%). RESULTS: Among all people with diabetes, 78% practiced self monitoring of blood glucose, and 25% were aware of the term "glycosylated hemoglobin" or "hemoglobin A one C" (HbA1c). In the last year, 72% of the subjects visited a health care provider for diabetes care at least once, 61% had their feet inspected at least once, and 61% received a dilated eye examination. Controlled for age and sex, the odds ratios (ORs) for insulin use were for self monitoring (OR [95% CI]; 4.0 [2.6-6.1]); having heard of HbA1c or receipt of a dilated eye examination (1.9 [1.4-2.5]); at least one visit to a provider (3.4 [1.9-7.2]); and feet inspected at least once (2.1 [1.5-2.9]). In addition, people <45 years, those who did not complete high school, and those without insurance coverage were high-risk groups for underuse of preventive care. Only 3% of insulin users and 1% of nonusers met all five of the American Diabetes Association standards in the previous year. CONCLUSIONS: Underuse of recommended preventive care practices is common among people with diabetes. PMID- 9727888 TI - Stratifying patients at risk of diabetic complications: an integrated look at clinical, socioeconomic, and care-related factors. SID-AMD Italian Study Group for the Implementation of the St. Vincent Declaration. AB - OBJECTIVE: The aim of this study was to identify subgroups of patients for whom the interactions among clinical, socioeconomic, and care-related factors determine a substantial increase in the risk of developing long-term diabetic complications. RESEARCH DESIGN AND METHODS: We performed a case-control study aimed at identifying and quantifying the risk factors for the development of major diabetic complications (eye, renal, and lower limb complications) in type 1 and type 2 diabetic patients. A total of 886 patients with renal, eye, or lower limb complications and 1,888 control subjects were enrolled in 35 diabetes outpatient clinics and 49 general practitioners' offices in 17 out of the 20 Italian regions. The main results were obtained using recursive partitioning and amalgamation (RECPAM), a technique that attempts to integrate the advantages of main effect logistic regression and tree-growing. RESULTS: The application of RECPAM led to the detection of important interactions involving clinical, socioeconomic, and care-related characteristics and allowed the identification of internally homogeneous subgroups characterized by a marked difference in the risk of developing major complications. In type 1 diabetic patients, the interaction between hypertension and smoking habits led to a dramatic increase in the complication risk, while in type 2 diabetic subjects, a poor compliance with visit scheduling was the most important predictor of complications. Furthermore, a marked difference in the risk profile was associated with patient characteristics (age, years of education, occupation). CONCLUSIONS: In the definition of the risk profile for each individual patient, socioeconomic status and level of education need to be taken under serious consideration, since they can determine a complication risk not dissimilar from hard clinical variables, such as hypertension and diabetes duration. Specific educational interventions, targeted to the socially disadvantaged strata of the population, need to be designed and implemented. PMID- 9727889 TI - Two-step islet autoantibody screening for risk assessment of type 1 diabetes in relatives. AB - OBJECTIVE: To examine the performance of islet cell antibodies (ICAs) and antibodies to glutamate decarboxylase (GADA), IA-2 (IA-2 antibody [IA-2A]), and insulin (insulin autoantibody [IAA]), alone and in combination, in assessing type 1 diabetes risk within type 1 diabetic families to identify a practical and effective screening strategy for predicting type 1 diabetes in relatives. RESEARCH DESIGN AND METHODS: ICA, GADA, IA-2A, and IAA were determined in 806 first-degree relatives participating in a prospective type 1 diabetes family study (median follow-up 6.17 years, range 0.6-8.3). The conferred risk of developing type 1 diabetes within 6 years was evaluated by Kaplan-Meier for each antibody marker, used alone or in combination. RESULTS: ICAs were detected in 3%, GADA in 5.1%, IA-2A in 2.5%, and IAA in 3.7% of relatives; > or =1 antibody markers were detected in 10.7% of relatives and > or =2 were detected in 1.9% of relatives. The risk of type 1 diabetes at 6 years was 1.5% in relatives with only 1 marker and 24.8% in relatives with > or =2 markers. As a practical and effective strategy for type 1 diabetes risk assessment in relatives, this study indicates a first-step screening based on GADA and IA-2A measurement--which identified 6.5% of relatives, including all who developed the disease, with a 6 year type 1 diabetes risk of 9.0%--followed by a second step based on ICA and IAA measurement in relatives with either GADA or IA-2A, which identified a total of 1.9% of all relatives as having > or =2 markers, and a 6-year risk of 24.8%, including 6 of 7 who developed type 1 diabetes. CONCLUSIONS: A two-step antibody screening, based first on GADA and IA-2A and then on ICA and IAA measurements in identified individuals, is likely to be a practical, sensitive, and effective strategy for predicting type 1 diabetes in first-degree relatives. PMID- 9727890 TI - Dissociation of microangiopathy and macroangiopathy in patients with type 2 diabetes. AB - OBJECTIVE: Although persistent hyperglycemia contributes greatly to the progression of diabetic micro- and macroangiopathy, microangiopathy progresses more rapidly than macroangiopathy in some type 2 diabetic patients, with the opposite being true in others. This study was conducted to identify factors responsible for such dissociation. RESEARCH DESIGN AND METHODS: Patients with proliferative diabetic retinopathy and a carotid intima-media thickness (IMT) level < or =1.0 mm were classified as the microangiopathy group (MIG); those with an IMT level >1.1 mm and without retinopathy or with background retinopathy were assigned to the macroangiopathy group (MAG). Only middle-aged patients, 50-69 years old, were included in this study. There were 54 patients in the MIG and 68 patients in the MAG. RESULTS: Patients in the MIG were significantly younger at the onset of diabetes, and those in the MAG had a significantly higher mean ratio of apoprotein (apo) B to apoAI. The percentage of patients with a family history of diabetes was significantly higher in the MIG. Maternal inheritance was common among these patients. Those with obesity, a family history of diabetes, and younger onset of hypertension were more common in the MAG. In the multiple logistic regression analyses, maternal inheritance and early onset of diabetes were independent risk factors for the acceleration of microangiopathy. A personal history of obesity and a family history of hypertension were independently related to the development of macroangiopathy. CONCLUSIONS: Our results suggest that patients with early onset and maternal inheritance of diabetes may have a high risk for the progression of diabetic microangiopathy, while patients with hyperlipidemia, a history of obesity, and a family history of hypertension seem prone to the development of atherosclerosis. PMID- 9727891 TI - Troglitazone use in insulin-treated type 2 diabetic patients. The Troglitazone Insulin Study Group. AB - OBJECTIVE: To determine the ability of troglitazone to reduce requirements for injected insulin while maintaining blood glucose levels in insulin-treated patients with type 2 diabetes. RESEARCH DESIGN AND METHODS: This 26-week double blind study with open-label extension included patients who had failed previous oral antidiabetic medication and took > or =30 but <150 U of insulin daily The 222 patients in the double-blind study received 200 or 400 mg troglitazone once daily or matching placebo. The primary end point was the proportion of patients meeting the target of > or =50% reduction in injected insulin and either a 15% reduction in fasting blood glucose or a blood glucose <7.8 mmol/l. Insulin dose was reduced 25% based on a study-specific algorithm whenever fasting blood glucose was reduced 5% from baseline. Also of interest were changes in insulin dose and HbA1c. The open-label extension included 173 patients. They received 200 mg of troglitazone with optional titration to 400 mg, and insulin dose was adjusted based on investigators' standards of care. Open-label measures were change in insulin dose, HbA1c, and fasting serum glucose (FSG). RESULTS: In the double-blind phase, 22 and 27% of the 200- and 400-mg troglitazone groups, respectively, reached target, compared with placebo (7%) (P < 0.01). Insulin dose reductions of 13 +/- 3, 30 +/- 3, and 41 +/- 3 U were observed for placebo, 200-, and 400-mg troglitazone groups, respectively HbA1c decreased 0.09 +/- 0.14% for placebo, 0.13 +/- 0.14% for 200 mg, and 0.41 +/- 0.14% for 400 mg (P < 0.05) troglitazone. In the open-label extension, troglitazone treatment resulted in >50% reduction from baseline in daily insulin dose and decreases in HbA1c of 1% and in FSG of >17%. CONCLUSIONS: Troglitazone decreases daily injected insulin dose requirements and improves glycemic control in insulin-treated patients with type 2 diabetes. PMID- 9727892 TI - Troglitazone in combination with sulfonylurea restores glycemic control in patients with type 2 diabetes. The Troglitazone Study Group. AB - OBJECTIVE: To determine if the combination of troglitazone (a peroxisome proliferator-activated receptor-gamma activator) and sulfonylurea will provide efficacy not attainable by either medication alone. RESEARCH DESIGN AND METHODS: There were 552 patients inadequately controlled on maximum doses of sulfonylurea who participated in a 52-week randomized active-controlled multicenter study. Patients were randomized to micronized glyburide 12 mg q.d. (G12); troglitazone monotherapy 200, 400, or 600 mg q.d. (T200, T400, T600); or combined troglitazone and glyburide q.d. (T200/G12, T400/G12, T600/G12). Efficacy measures included HbA1c, fasting serum glucose (FSG), insulin, and C-peptide. Effects on lipids and safety were also assessed. RESULTS: Patients on T600/G12 had significantly lower mean (+/- SEM) FSG (9.3 +/- 0.4 mmol/l; 167.4 +/- 6.6 mg/dl) compared with control subjects (13.7 +/- 0.4 mmol/l; 246.5 +/- 6.8 mg/dl; P < 0.0001) and significantly lower mean HbA1c (7.79 +/- 0.2 vs. 10.58 +/- 0.18%, P < 0.0001). Significant dose-related decreases were also seen with T200/G12 and T400/G12. Among patients on T600/G12, 60% achieved HbA1c < or =8%, 42% achieved HbA1c < or =7%, and 40% achieved FSG < or =7.8 mmol/l (140 mg/dl). Fasting insulin and C peptide decreased with all treatments. Overall, triglycerides and free fatty acids decreased, whereas HDL cholesterol increased. LDL cholesterol increased slightly, with no change in apolipoprotein B. Adverse events were similar across treatments. Hypoglycemia occurred in 3% of T600/G 12 patients compared with <1% on G12 or troglitazone monotherapy CONCLUSIONS: Patients with type 2 diabetes inadequately controlled on sulfonylurea can be effectively managed with a combination of troglitazone and sulfonylurea that is safe, well tolerated, and represents a new approach to achieving the glycemic targets recommended by the American Diabetes Association. PMID- 9727893 TI - Troglitazone reduces plasma leptin concentration but increases hunger in NIDDM patients. AB - OBJECTIVE: Troglitazone, which improves peripheral insulin resistance of experimental diabetic animals and diabetic patients, affects ob gene expression in the adipose tissue of rodents. The present study was undertaken to examine a hypothesis that clinical administration of troglitazone may reduce circulating leptin levels and affect eating behavior in NIDDM patients. RESEARCH DESIGN AND METHODS: Troglitazone was administered at a dosage of 200 mg twice daily for 12 weeks in 20 poorly controlled NIDDM patients. Chronological changes in glycemic control, serum lipids, immunoreactive leptin (IRL) levels, and BMI were measured. Body fat weight was also assessed by bioelectric impedance. RESULTS: Troglitazone significantly decreased fasting plasma glucose, serum immunoreactive insulin, and HbA1c levels. Serum levels of IRL and triglyceride were significantly reduced by troglitazone administered for 4, 8, and 12 weeks. Troglitazone administration significantly increased the BMI in NIDDM patients, and two-thirds of the patients complained of increased hunger after the start of troglitazone administration. CONCLUSIONS: Troglitazone significantly reduces circulating leptin levels at clinical doses. It may affect the eating behavior of poorly controlled NIDDM patients through the improvement of glycemic control and/or the reduction of circulating leptin. PMID- 9727894 TI - Comparison of percent total GHb with percent HbA1c in people with and without known diabetes. AB - OBJECTIVE: To directly compare results obtained using an ion-exchange high performance liquid chromatography (HPLC) HbA1c method used in the Diabetes Control and Complications Trial with two different affinity chromatography methods in which "total GHb" is determined. RESEARCH DESIGN AND METHODS: Blood was obtained from a large number of people with and without known diabetes. The specimens were divided and assayed for HbA1c and for total GHb. Total GHb was determined using a semi-automated gravity-elution boronate affinity chromatography method and an automated boronate affinity HPLC method. The results obtained with the two methods were also compared. RESULTS: In subjects without known diabetes, the mean percentage HbA1c and the range of values were similar to the total GHb values in the same subjects when assayed using the semi-automated affinity gravity-elution method (mean 5.2 +/- 0.4 and 5.1 +/- 0.4% [SD], respectively). With the affinity HPLC method, results were 5.3 +/- 0.4%. The similarity in results was surprising. However, analysis of the data suggests that a large proportion of the material in the HbA1c fraction measured using this ion exchange HPLC method is not GHb, as pointed out by others. Although the results were similar in people without known diabetes, in the people with diabetes, the incremental increase was approximately 25% greater for the total GHb when compared with the increase in HbA1c. When corrected for the non-GHb being measured by the HbA1c method, it can be calculated that approximately 40% more GHb is measured using affinity chromatography over the entire range of GHb values. CONCLUSIONS: The similarity in the mean and range of percent HbA1c and in percent total GHb using these different methods can be attributed to two factors: 1) the HbA1c ion-exchange method measures only approximately 50-60% of the total GHb present, and 2) approximately 40-50% of the material being measured in the HbA1c fraction is not GHb, i.e., offsetting factors fortuitously resulted in values similar to the more specific affinity methods. The greater incremental increase in percent total GHb compared with percent HbA1c in people with diabetes can be attributed to the greater amount of GHb being measured with the affinity methods. PMID- 9727895 TI - Microdialysis of glucose in subcutaneous adipose tissue up to 3 weeks in healthy volunteers. AB - OBJECTIVE: To measure possible changes in dialysate glucose concentrations over time, to validate the diffusional model for glucose transport from tissue to the probe, and to evaluate the actual glucose concentration in adipose tissue. RESEARCH DESIGN AND METHODS: Glucose concentrations in the subcutaneous adipose tissue of five healthy subjects (age 25 +/- 2.7 years, BMI 23.2 +/- 2.3 kg/m2 [mean +/- SD]) were measured by the microdialysis technique and compared with blood glucose. We applied microdialysis probes with hollow fibers of various membrane length (10-35 mm), used eight perfusion flow rates (0.5-20 microl/min), and perfused four glucose solutions (0.0, 2.8, 8.3, 11.1 mmol/l). RESULTS: After implantation, a substantial decrease in glucose recovery to the lowest value of 26 +/- 10% of the final plateau value was noted during the first few hours (n = 4). Recovery increased and stabilized after 5-9 days at 84.0 +/- 7.4% of capillary blood glucose when a flow rate of 0.5 microl/min was applied. According to the zero net-flux method, the glucose concentration in equilibrium, Cequi, with the surrounding tissue can be obtained. This concentration also decreases; however, 1 h after recovery, Cequi increases again over 1 or 2 days to a stable value that is not significantly different from the measured capillary blood glucose (P < 0.05). Using various perfusion flow rates and probes (membrane length 10-35 mm), it is shown that diffusion is the rate-limiting process for glucose transport through tissue. CONCLUSIONS: Insertion of the microdialysis probes causes damage to the adipose cells and the vascular bed around the probe. Glucose recovery decreases because of a lower blood supply. In 5-9 days, glucose recovery increases; apparently, this time is needed to repair the microstructure of tissue around the probe. After stabilization of the recovery, no loss of probe permeability, which is due to biocompatibility problems, was seen. The change during the 2 days in equilibrium concentration is probably caused by an inflammation reaction that consumes glucose around the probe. The individual increase in recovery during the 1st days after probe insertion until a stable plateau value is reached (flow rate >0 microl/min) is complicated for short-term clinical glucose measurements in adipose tissue. After stabilization, the mean equilibrium concentration of all subjects was equal to the mean capillary blood glucose concentration. Therefore, we conclude that capillary blood glucose concentration probably is the driving force for diffusion through the capillary wall into the probe and is not some interstitial concentration. PMID- 9727897 TI - Sympathetic function test of vasoconstrictor changes in foot arteries in diabetic patients. AB - OBJECTIVE: We studied the relationship between vasoconstrictor changes in foot arteries (pedal, metatarsal, and digital arteries) and autonomic neuropathy in diabetic patients to estimate the degrees of sympathetic dysfunction. RESEARCH DESIGN AND METHODS: Sixty-two patients and nineteen age-matched control subjects were studied. The resistance index (RI) and pulsatility index (PI) were measured as vascular hemodynamic parameters using Doppler sonography, and the increases in these hemodynamic parameters (%RI and %PI) from rest to a deep breath were measured as indexes of the degrees of sympathetic vasoconstrictor function. Cardiovascular autonomic function tests (AFTs) were performed and the score was compared to %RI and %PI values obtained. RESULTS: Of the 62 diabetic patients, 51 had various degrees of autonomic neuropathy. Both %RI and %PI in the diabetic patients were significantly less than those in the control subjects for all foot arteries tested (all P < 0.001). There were strongly inverse correlations between the %RI and %PI of foot arteries and the AFT score (r = -0.556 to -0.846, P < 0.0001). The %RI of the digital artery was the most strongly correlated with AFT score (r = -0.846, P < 0.0001) among foot arteries tested. The abnormality of sympathetic vasoconstriction was detectable in the majority of the diabetic patients with the early phase of autonomic neuropathy (%RI: 89.5%; %PI: 94.5%). CONCLUSIONS: We conclude that the %RI of the digital artery is a useful and reliable sympathetic function test of early phase in diabetic patients. PMID- 9727896 TI - Irreversibility of the defect in glycogen synthase activity in skeletal muscle from obese patients with NIDDM treated with diet and metformin. AB - OBJECTIVE: To assess the reversibility of the defect in glycogen synthase (GS) activity in skeletal muscle from obese patients with NIDDM treated with a hypocaloric diet and metformin. RESEARCH DESIGN AND METHODS: Eighteen obese patients newly diagnosed with NIDDM were included in a randomized placebo controlled double-blind parallel group trial and followed for 3 months. Euglycemic-hyperinsulinemic clamp including indirect calorimetry and biopsy of m. vastus lateralis was performed before and after treatment with a hypocaloric diet plus metformin or placebo. The patients were studied at basal, low, and high insulin concentrations. RESULTS: The impaired GS activity in muscle biopsies was not reversed either by acute normalization of glycemia (for 8 h) or by chronic reduction of hyperglycemia by diet plus metformin. In both treatment groups, comparable effects on glycemic control and weight loss were found together with marked insulin suppression of nonesterified fatty acids and increased glucose oxidation. Total glucose disposal at euglycemic-hyperinsulinemic clamp increased significantly in the metformin group by 25% at high insulin level (259 +/- 31 vs. 207 +/- 21 mg x m(-2) x min(-1), P < 0.05). An insignificant increase by 13% was found in the placebo group. There were no significant changes in nonoxidative glucose metabolism. GS activity and glucose utilization showed no significant differences between the two treatment groups when regression coefficients, expressed as incremental changes by increments of insulin, were compared. CONCLUSIONS: Defective GS activity in obese NIDDM patients is not secondary to hyperglycemia. Metformin and diet had no significant influence on GS activity. The added effect of metformin to that of a hypocaloric diet in improving insulin stimulated glucose utilization is marginal when blood glucose reduction is obtained by weight loss. PMID- 9727898 TI - Biologic material in needles and cartridges after insulin injection with a pen in diabetic patients. AB - OBJECTIVE: To evaluate the frequency of non-inert material, including cells, in needles and cartridges after insulin injection with pen-like devices in diabetic patients. RESEARCH DESIGN AND METHODS: A prospective study was conducted in 120 insulin-treated diabetic patients who used pen-like devices. The patients, 46 women and 74 men, were 20-77 years old; 60% had type 1 diabetes, and 38% were overweight. Duration of diabetes ranged from 1 month to 40 years, and insulin therapy ranged from 1 month to 30 years. Insulin injection was performed by a trained nurse, using the patient's usual pen and cartridge. A cytopathological examination was performed on the material obtained from the needle and found in the cartridge after centrifugation. All slides were read by a single investigator. RESULTS: In 62% of the patients, non-inert material was found, including squamous (32%) and epithelial (58%) cells. Biologic material was found in 30% of the needles and 58% of the cartridges, and in both needle and cartridge in 25% of the population. Biologic material was found more frequently in patients who had a longer duration of diabetes, who were treated with insulin for a longer time, and who performed injection in the thighs or upper arms (P < 0.05). In multivariate analysis, the presence of biologic material was associated with the duration of diabetes (R2 = 0.09; P < 0.01). CONCLUSIONS: Our data suggest that biologic material can be trapped in the delivery system, including the cartridge, after an insulin injection with a pen-like device. Our results emphasize the strict need for individual use of insulin delivery systems, including cartridges and nonrefillable pens, especially in clinics and hospitals. PMID- 9727899 TI - Effect of hypoglycemia on beta-adrenergic sensitivity in normal and type 1 diabetic subjects. AB - OBJECTIVE: The purpose of this study was to assess the potential role of reduced tissue sensitivity to catecholamines in the pathogenesis of hypoglycemia unawareness in patients with type 1 diabetes. RESEARCH DESIGN AND METHODS: The effect of a single episode of hypoglycemia on beta-adrenergic sensitivity was studied in 10 type 1 diabetic patients with apparently normal awareness of hypoglycemia (age 29 +/- 5 years, diabetes duration 13 +/- 8 years, HbA1c 7.3 +/- 0.9%) and 10 age-matched healthy control subjects. Beta-adrenergic sensitivity was measured with the isoproterenol test after a hyperinsulinemic euglycemic clamp and after a hyperinsulinemic hypoglycemic clamp. Beta-adrenergic sensitivity was expressed as the dose of intravenous isoproterenol that increased the heart rate by 25 beats/min (IC25). RESULTS: During hypoglycemia, diabetic subjects had an impaired plasma epinephrine response compared with that of the control subjects (16.7 +/- 5.0 vs. 40.1 +/- 6.8 ng/ml, P = 0.02). In control subjects, the IC25 was lower after hypoglycemia than after euglycemia (0.83 +/- 0.22 vs. 1.13 +/- 0.21 microg, P = 0.02) indicating an increase in beta adrenergic sensitivity. In diabetic subjects, on the other hand, the IC25 was greater after hypoglycemia than after euglycemia (1.00 +/- 0.26 vs. 0.65 +/- 0.14 microg, P = 0.04), indicating a decrease in beta-adrenergic sensitivity. CONCLUSIONS: In normal subjects, a single episode of hypoglycemia increases beta adrenergic sensitivity. In diabetic subjects, in contrast, hypoglycemia reduces beta-adrenergic sensitivity. These results provide evidence that in type 1 diabetic patients, some maladaptation of tissue sensitivity to catecholamines contributes to the development of hypoglycemia unawareness. A unifying hypothesis is presented for the pathogenesis of hypoglycemia unawareness in type 1 diabetic patients incorporating the concepts of both a reduced catecholamine response and reduced adrenergic sensitivity PMID- 9727900 TI - Relationship of blood thromboxane-B2 (TxB2) with lipid peroxides and effect of vitamin E and placebo supplementation on TxB2 and lipid peroxide levels in type 1 diabetic patients. AB - OBJECTIVE: To study the effect of vitamin E supplementation on platelet hyperaggregability in type 1 diabetic patients. RESEARCH DESIGN AND METHODS: Written informed consent according to the Institutional Review Board on Human Experimentation guidelines was obtained from diabetic patients (n = 29) and their age-matched normal siblings (n = 21) to participate in this study. Diabetic patients were supplemented with DL-alpha-tocopherol (vitamin E) capsule (orally, 100 IU/day) or placebo for 3 months in a double-blind clinical trial. Alternate diabetic patients were assigned to vitamin E or placebo during regular visits to the clinic. Fasting blood was collected from each diabetic patient before the start and after the vitamin E or placebo supplementation. Platelet aggregability was assessed by competitive enzyme-linked immunosorbent assay of the blood TxB2 (a stable thromboxane metabolite). Plasma vitamin E and MDA (malondialdehyde, a product of lipid peroxidation) was assessed by high-performance liquid chromatography. Data were analyzed statistically on 12 diabetic patients on vitamin E and 12 on placebo supplementation. RESULTS: Diabetic patients (n = 29) had 62% higher (P < 0.05) levels of TxB2 and 15% higher levels (P < 0.05) of MDA in comparison to normal subjects (n = 21). Plasma TxB2 levels had a significant correlation with MDA levels (r = 0.45, P < 0.02) but not with the HbA1 (r = 0.08), glucose (r = -0.13), duration of diabetes (r = -0.04), or age (r = 0.12) of diabetic patients. Vitamin E supplementation lowered MDA levels by 30% (P < 0.04), TxB2 levels by 51% (P < 0.03), and triglyceride levels by 22% (P < 0.04) in diabetic patients. There were no differences in these parameters before versus after placebo supplementation. CONCLUSIONS: The elevated blood level of TxB2 (hyperaggregability of platelets) is significantly related to the level of lipid peroxidation products (oxidative stress) in type 1 diabetic patients. Supplementation of modest doses of vitamin E (100 IU/day) significantly lowers blood TxB2 and lipid peroxidation products levels in type 1 diabetic patients. PMID- 9727901 TI - Plasma lipoprotein(a) levels are not influenced by glycemic control in type 1 diabetes. AB - OBJECTIVE: To determine the influence of glycemic control improvement with intensive therapy on lipoprotein(a) [Lp(a)] concentrations in type 1 diabetic patients. RESEARCH DESIGN AND METHODS: A total of 105 poorly controlled type 1 diabetic patients (60 men, 45 women) without diabetic complications participated in a longitudinal study performed in a tertiary referral center, to compare lipid, lipoprotein, and Lp(a) levels before and after 3 months of intensive therapy with multiple insulin doses. Lp(a) levels were measured by the Terumo method. Differences between the two periods were assessed by the paired t test and Wilcoxon's test. RESULTS: After 3 months of intensive therapy, all patients exhibited improved glycemic control. HbA1c decreased from 8.9 +/- 2.4 to 6.5 +/- 1.6% (P < 0.0001), being < or =6% in 47% of patients. However, although a more favorable lipoprotein profile was obtained, no changes in Lp(a) concentrations were observed in the whole group of patients (16.7 +/- 17.3 vs. 17.2 +/- 17.7 mg/dl) or in patients with baseline Lp(a) levels above 30 mg/dl (47.1 +/- 14.8 vs. 47.4 +/- 18.9 mg/dl) or below 30 mg/dl (9.6 +/- 7.3 vs. 10.2 +/- 6.7 mg/dl). In addition, patients reaching HbA1c < or =6 or >6% presented similar Lp(a) levels (19.7 +/- 18.0 vs. 15.0 +/- 17.4 mg/dl), and changes in Lp(a) did not correlate with those observed in HbA1c. CONCLUSIONS: These data demonstrate that the improvement of glycemic control does not influence plasma Lp(a) concentrations in type 1 diabetic patients independently of baseline Lp(a) levels and the degree of glycemic control. PMID- 9727902 TI - Urinary albumin excretion rate and glomerular filtration rate in single-kidney type 2 diabetic patients. AB - OBJECTIVE: To evaluate the urinary albumin excretion rate (UAER) and the glomerular filtration rate (GFR) of single-kidney type 2 diabetic patients (SKD) and of single-kidney non-diabetic patients (SKN). RESEARCH DESIGN AND METHODS: Patients who had only one kidney for at least 5 years, with no renal disease or hypertension at the time of the nephrectomy and with no calculus or systemic disease at the time of the evaluation, were included in this controlled cross sectional study A total of 20 SKD (8 men, age 62 +/- 9 years; diabetes duration 8.5 +/- 7 years), 17 SKN (2 men, age 57 +/- 13 years), and 184 type 2 diabetic patients who were matched to the single-kidney diabetic group for age, sex, and BMI were studied. UAER was measured by immunoturbidimetry in timed 24-h sterile urine, and GFR was determined by the 51Cr-EDTA single-injection method. RESULTS: SKD patients presented a higher proportion (8 of 20, 40%) of microalbuminuria (UAER 20-200 microg/min) than SKN patients (3 of 17, 17.6%) and type 2 diabetic patients (37 of 184, 20%). SKD patients presented a higher proportion of macroalbuminuria (UAER >200 microg/min; 6 of 20, 30%) than SKN patients (1 of 17, 6%) but were similar to type 2 diabetic patients (43 of 184, 23%). The GFRs of normoalbuminuric SKN (71.7 +/- 21.4 ml x min(-1) x 1.73 m(-2)) and SKD patients (73.0 +/- 21.5 ml x min(-1) x 1.73 m(-2)) were similar but higher than the one kidney GFR (GFR / 2) of the age-, sex-, and BMI-matched normal individuals (50.5 +/- 9.0 ml x min(-1) x 1.73 m(-2)) and normoalbuminuric type 2 diabetic patients (54.0 +/- 11.6 ml x min(-1) x 1.73 m(-2)). CONCLUSIONS: Increased GFR related to single-kidney status confers an increased risk of developing renal disease in the presence of diabetes. PMID- 9727903 TI - Hyperglycemia is a factor for an increase in serum ceruloplasmin in type 2 diabetes. AB - OBJECTIVE: To examine if there is a correlation between high blood glucose and serum ceruloplasmin (Cp) levels. RESEARCH DESIGN AND METHODS: Serum Cp levels were measured in 637 patients with type 2 diabetes (all type 2 diabetes group). For the follow-up type 2 diabetes group, 161 patients who had not had any changes in their situation during the last year that are known to influence serum Cp levels were reexamined 1 year later. The control group was composed of 158 healthy individuals. Serum Cp and blood HbA1c levels were measured by radial immunodiffusion and high-performance liquid chromatography assays, respectively. RESULTS: Serum Cp levels in the all type 2 diabetes group were significantly higher than those in the control group (P < 0.0001), although the serum Cp levels did not correlate with the blood HbA1c levels in the all type 2 diabetes group (r = 0.055, P = 0.351). Then we evaluated those factors (delta-log Cp and delta HbA1c) in the follow-up type 2 diabetes group to minimize changes from the genetic differences and to exclude any known factors influencing serum Cp levels. This indicated that the delta-HbA1c had a positive correlation to the delta-log Cp (r = 0.304, P < 0.0001). CONCLUSIONS: A persistent high blood glucose (namely HbA1c) is associated with an increase in serum Cp levels over 1 year. PMID- 9727904 TI - Meal-generated oxidative stress in type 2 diabetic patients. AB - OBJECTIVE: Free radical production has been reported to be increased in diabetic patients and to be involved in the pathogenesis of diabetic complications. In this study, a standardized meal was administered to 10 type 2 diabetic patients and 10 healthy matched normal subjects to evaluate its effects on plasma oxidative stress generation. RESEARCH DESIGN AND METHODS: In diabetic patients, at baseline and after the meal, plasma malondialdehyde (MDA), vitamin C, protein SH groups, uric acid, vitamin E, and total plasma radical-trapping parameter, which evaluates plasma antioxidant capacity due to known and unknown antioxidants present in the plasma as well as their mutual cooperation, were measured. RESULTS: After the meal, plasma MDA and vitamin C increased, while protein SH groups, uric acid, vitamin E, and total plasma radical-trapping parameter decreased more significantly in the diabetic subjects than in control subjects. CONCLUSIONS: This finding shows that in the absorptive phase, free radicals are produced in diabetic patients. Since plasma glucose, but not insulin, rose significantly more in diabetic subjects than in control subjects, hyperglycemia may play an important role in the generation of postprandial oxidative stress in diabetic patients. PMID- 9727905 TI - Increased left ventricular mass in normotensive type 1 diabetic patients with diabetic nephropathy. AB - OBJECTIVE: Diabetic nephropathy increases the risk of premature cardiovascular disease and sudden death, particularly in type 1 diabetic patients. One possible mechanism for this risk may be left ventricular hypertrophy. In our study, we aimed to evaluate left ventricular structure and function in normotensive type 1 diabetic patients with and without nephropathy. RESEARCH DESIGN AND METHODS: M mode and Doppler echocardiography was performed in 17 type 1 diabetic patients with nephropathy (albuminuria [median (range)], 345 (135-2,846) mg/24 h) and compared with 34 normotensive, normoalbuminuric (10 [3-30] mg/24 h) type 1 diabetic patients matched for arterial blood pressure (mean +/- SD) ([134/77] +/- [13/7] vs. [129/78] +/- [12/7] mmHg), age (40 +/- 11 vs. 42 +/- 10 years), duration of diabetes (28 +/- 7 vs. 28 +/- 6 years), and BMI (24.2 +/- 4.2 vs. 24.6 +/- 2.4 kg/m2). RESULTS: Left ventricular mass (LVM) index was significantly higher in patients with nephropathy compared with patients with normoalbuminuria (100.8 +/- 10.3 vs. 88.2 +/- 21.0 g/m2, respectively; P = 0.02). Greater ventricular septum width was demonstrated in the nephropathic group compared with the control group (9.4 +/- 1.0 vs. 8.2 +/- 1.3 mm, respectively; P = 0.002). No significant difference in posterior wall thickness was apparent. The nephropathic group tended to have reduced diastolic function (E/A ratio, 1.2 +/- 0.3 vs. 1.4 +/- 0.4; P = 0.09). Fractional shortening was normal and about the same in the two groups. The groups did not differ with respect to serum creatinine or hemoglobin, while metabolic control (assessed by HbA1c) and plasma renin and prorenin levels were elevated in the nephropathic group compared with the normoalbuminuric group. CONCLUSIONS: A blood pressure-independent increase in LVM may contribute to the increased cardiac morbidity and mortality in normotensive type 1 diabetic patients with diabetic nephropathy. Glycemic abnormalities and activation of the renin-angiotensin system may lead to the ventricular enlargement. PMID- 9727906 TI - Association of NIDDM and hearing loss. AB - OBJECTIVE: To evaluate the association of NIDDM with hearing loss in a large population-based study. RESEARCH DESIGN AND METHODS: Data from population-based longitudinal studies of aging conducted in Beaver Dam, Wisconsin, were used in these analyses. Hearing thresholds were determined by pure-tone air- and bone conduction audiometry performed by trained technicians following American Speech Language-Hearing Association specifications. Hearing loss was defined as the pure tone average of the frequencies 500, 1,000, 2,000, and 4,000 Hz greater than 25 decibels hearing level in the worse ear. Diabetes status was determined by self report of physician-diagnosed diabetes or by elevated glucose or glycated hemoglobin levels at examination. RESULTS: Of 3,571 study participants, 344 were classified as having NIDDM. Subjects with NIDDM were more likely to have a hearing loss than were subjects without diabetes (59 vs. 44%). After results were adjusted for age, this difference was not statistically significant. After individuals with hearing loss patterns inconsistent with presbycusis were excluded, there was an association between NIDDM and hearing loss when controlling for potential confounders (odds ratio [OR] 1.41, 95% CI 1.05-1.88). There was no association between duration of diabetes or glycemic control and hearing loss. Individuals with NIDDM and nephropathy were more likely to have a hearing loss than were those with NIDDM but no nephropathy (OR 2.28, 95% CI 1.04 5.00). CONCLUSIONS: These data are suggestive of a weak association between NIDDM and hearing loss. PMID- 9727908 TI - Consensus development conference on the diagnosis of coronary heart disease in people with diabetes: 10-11 February 1998, Miami, Florida. American Diabetes Association. PMID- 9727907 TI - Increased familial history of arterial hypertension, coronary heart disease, and renal disease in Brazilian type 2 diabetic patients with diabetic nephropathy. AB - OBJECTIVE: To evaluate whether there is a familial association of arterial hypertension, coronary heart disease, renal disease, and stroke with diabetic nephropathy RESEARCH DESIGN AND METHODS: There were 115 outpatients and 34 patients with end-stage renal disease treated by hemodialysis (61 men, age range 41-81 years) and having at least one sibling with type 2 diabetes studied. The positive or negative history of siblings (n = 765) was assessed by a standard questionnaire. The urinary albumin excretion rate (UAER) was measured by radioimmunoassay in 24-h sterile urine (three samples). The subjects were grouped as normoalbuminuric (UAER <20 microg/min, n = 59), microalbuminuric (UAER 20-200 microg/min, n = 35), macroalbuminuric (UAER >200 microg/min, n = 21), and end stage renal disease (n = 34). RESULTS: Patients with microalbuminuria, macroalbuminuria, or end-stage renal disease had an increased prevalence of sibling history of arterial hypertension (33.2, 37.3, and 33.8 vs. 23.4%, P < 0.001) and coronary heart disease (15.2, 17.0, and 19.4 vs. 10.2%, P = 0.044) compared with the normoalbuminuric group. The renal disease history was increased only in the siblings of patients with macroalbuminuria or end-stage renal disease (12.8 and 15.6 vs. 7.6 and 6.1%, P = 0.005). The presence of sibling arterial hypertension strongly increases the prevalence of sibling renal and coronary heart disease independent of patient renal status. CONCLUSIONS: There is an association of diabetic nephropathy and sibling history of arterial hypertension and renal and coronary heart disease in type 2 diabetic patients. These associations are not independent, and arterial hypertension may be their main determining factor. PMID- 9727910 TI - Assessing dependency in insulin-treated patients with diabetes supported by a community nursing service. PMID- 9727909 TI - International Diabetes Federation meeting, 1997: nephropathy, retinopathy, and glycation. PMID- 9727911 TI - Transient hypoglycemic hemiparesis in children with IDDM. PMID- 9727912 TI - No differences in attentional functioning between type 1 diabetic patients with and without a history of severe hypoglycemia. PMID- 9727913 TI - A novel missense mutation in the homeodomain of the hepatocyte nuclear factor 1alpha/maturity-onset diabetes of the young 3 in a Japanese early-onset type 2 diabetic patient and time-course of glucose-stimulated insulin secretion. PMID- 9727914 TI - An epitaph for sulfated insulin: immunologic profile of the last patients as they are switched from sulfated beef to human insulin. PMID- 9727915 TI - Is diabetic ketoacidosis a cause of meningeal syndrome? Case report. PMID- 9727916 TI - Organospecific lymph node enlargement in autoimmune polyglandular syndrome. PMID- 9727917 TI - Insulin analog lispro decreases insulin resistance and improves glycemic control in an obese patient with insulin-requiring type 2 diabetes. PMID- 9727918 TI - Type 2 diabetes in adults with pseudopseudohypoparathyroidism. Case report. PMID- 9727919 TI - High prevalence of albuminuria among African-Americans with short duration of diabetes. PMID- 9727920 TI - Use of HbA1c in screening for diabetes. PMID- 9727921 TI - Assessing the utility of glycated hemoglobin. PMID- 9727922 TI - Agreement between old and new diagnostic criteria in postpartum testing of women with gestational diabetes. PMID- 9727923 TI - Three-dimensional reconstruction of types IV and V midfacial defects by free rectus abdominis myocutaneous (RAM) flap. AB - Extensive midfacial defects after ablative surgery constitutes a challenging problem for reconstructive surgeons. Particularly for types IV and V midfacial defects, provision of missing bony support and obliteration of the maxillary cavity defects require microsurgical free tissue transfers. In the last three years, four patients have undergone total maxillectomy for midfacial tumours and the postmaxillectomy defects were three-dimensionally repaired with free rectus abdominis muscle flap and skin graft or myocutaneous flaps. Obliteration of maxillary cavity defects and orbital support were achieved with this type of free flap. The least follow-up period of the patients is one year and slight ectropion, later corrected, was seen in two patients. In this study, the free rectus abdominis myocutaneous (RAM) flap, with its long vascular pedicle and availability of various skin paddle designs and muscle bulk, is presented in treatment of extensive midfacial defects. In spite of initial overcorrection of contour, the denervated rectus abdominis muscle gradually atrophies, resulting in loss of contour. The muscle bulk fills the cavity defect, but, in order to achieve good facial contour, it is necessary to support the bony skeleton with some material. PMID- 9727924 TI - Temporomandibular joint reconstruction with free microvascular transfer of the metatarsophalangeal joint: a case report. AB - Free microvascular transfer of the second metatarsophalangeal joint was performed for the treatment of temporomandibular joint ankylosis in a 15-year-old male patient. The result is excellent in one-year follow-up. The technique seems to be a good alternative to the problem in selected patients. PMID- 9727925 TI - Reconstruction of mandible with fibula free flap. AB - The fibula can be used as a donor for a free flap for mandible reconstruction. It has the advantages of low donor site morbidity, consistent shape, ample length, and distant location to enable a two-team approach, allowing multiple osteotomies because of its periosteal circulation. It can be raised with a skin island for composite tissue reconstruction. Eight segmental mandibular defects (average 11.62 cm) were reconstructed following resection for tumour. Six defects consisted of bone alone and the other two had only a small amount of associated intraoral soft-tissue loss. Two patients underwent primary reconstruction. We performed two or three osteotomies on each graft and used miniplates and wires for bone fixation. The flaps survived in all patients. All osteotomy sites healed primarily. The aesthetic result of reconstruction was satisfactory. PMID- 9727926 TI - Vascularised bone graft applications in upper extremity problems. AB - Between 1990 and 1996, 16 cases of bone defects were treated by vascularised bone grafting by the authors. Free vascularised fibula was used in 10 cases and one free iliac crest graft was used for upper extremity bone defects. Four vascular pedicled first metacarpal bone and one radial styloid bone were used for scaphoid nonunion. Average follow-up was 26 months (6-78 months) and success rate was 94%. We recommend vascularised bone grafts in the upper extremity when there is risk of infection; the defect is greater than five centimeters when the forearm rotation is unlimited. The avascularity of the scaphoid pseudarthrosis must be verified radiologically or through magnetic resonance imaging. This technique should only be used when other reconstructive techniques are unlikely to succeed. PMID- 9727928 TI - Venous free flaps for reconstruction of skin defects of the hand. AB - Twenty-one cases of skin defects of the hand were treated with venous flaps. According to type, nine flaps were arterialised flaps (A-A), five were (A-V), and seven were (V-V) type. Venous flaps can be used up to 8 x 3 cm in size or even bigger if the number of veins anastomosed is increased. The main advantage of venous flaps is that they can also be used for simultaneous reconstruction of circulation in digits. PMID- 9727927 TI - Upper extremity salvage procedure with flow-through free flap transfer taken from the amputated part. AB - An alternative free flap technique in a patient presenting with an incomplete amputation in the right cubital region, resulting from a gunshot wound, is introduced. The defect area was repaired using a flow-through fasciocutaneous free flap, which, when following a cubical or popliteal region amputation, is a suitable option for revascularisation and salvage of the extremity. PMID- 9727929 TI - Anomalous digital nerve loop: a case report. AB - In this case, a digital nerve loop was observed during the dissection of the digital nerves to the third web space; the common digital artery was passing through this loop. This rare case is presented and its importance is discussed. PMID- 9727930 TI - Combined treatment of digital degloving injuries: a case report. AB - In a digital degloving injury of four fingers, a combined treatment of tripedicle dermal flap, free hemipulp web flap, and skin graft was used. The functional and cosmetic results of the hand were satisfactory. The patient had an excellent sensory recovery in the free hemipulp web flap and did not complain of any disability from the donor site. PMID- 9727931 TI - Reconstruction of soft tissue and bone defects in lower extremity with free flaps. AB - This article reports our experiences treating soft tissue and bone defects in the lower extremity with free flaps. One of the most common causes for lower extremity wounds are high-energy injuries. These kinds of injuries contain soft tissue and bone defects beyond neurovascular complications. The rate of infection and nonunion is very high in these injuries. Between 1988 and 1996, we applied 33 flaps to 28 patients. The numbers and kinds of the free flaps are as follows: 12 latissimus dorsi, nine radial forearm, seven lateral arm, three vascularised fibula grafts with skin, one gracilis, and one medial plantar flap. Three free flaps were lost (12%). The success rate is 88%. The advantage of free flaps is that they allow the reconstruction of the large defects in one-session operations. Thus, they decrease the rate of infection and increase that of nonunion. The patient returns to his active life at an earlier stage. PMID- 9727932 TI - Reconstruction of foot defects due to mine explosion using muscle flaps. AB - Landmine explosions bring a formidable challenge to both patients and reconstructive surgeons. Free tissue transfer is the only method of repairing such extensive soft tissue defects of the foot after serial debridements. Sixty five consecutive free muscle flap transfers were performed in 54 patients who had foot defects involving soft tissue and bone due to mine explosions. Although posttraumatic vessel disease had complicated most of the cases, overall flap survival rate was 83%. Each patient was ambulatory. Ulceration in long-term period was seen in only one patient. Eighty-five percent of patients with successful bone reconstruction and 41.6% of patients without adequate bone replacement demonstrated normal weightbearing in footprints and gait analysis. Free muscle flaps with split thickness skin graft and bone replacement are recommended for the reconstruction of such devastating wounds. PMID- 9727933 TI - Partial necrosis of an amputated penis following replantation in a heavy smoker: a case report. AB - A case of penile amputation in a heavy smoker who was treated by microsurgical replantation is presented. The penis was cut by a kitchen knife and the ischaemia time until revascularisation was 3 hours. Following successful microneurovascular repair the penis survived. However, a 3 cm portion of the ventral segment, including the corresponding urethral segment, sloughed. The adverse effects of nicotine on vascular anastomotic network in heavy smokers are well known and may be the cause of the partial necrosis in this case. PMID- 9727934 TI - Evaluation of the improvement of sensibility after primary median nerve repair at the wrist. AB - In the Istanbul Hand Surgery and Microsurgery Centre, between 1991-1996, 28 out of 54 patients who had clean-cut median nerve laceration at the wrist level were evaluated in a detailed manner regarding the improvement of sensibility following primary repair. Semmes-Weinstein monofilament and vibration tests form the group of threshold tests, and static and dynamic two-point discrimination tests and the Moberg pick-up test form the group of functional tests which were applied to the patients. Follow-up was from 1 to 5 years. Moreover, subjective findings such as cold intolerance, pain, and paraesthesia were also evaluated. All the results obtained were evaluated in relation to the SO-S4 sensibility improvement criterion which Waylett-Rendall recommended. The following results were obtained: S4 in 35.7% of the patients, S3+ in 28.5%, S3 in 14.2%, S2+ in 7.14%, S2 in 10.7%, and S1 in 3.5%. In our opinion the most important reason for the high rate of success was the fact that we worked with a highly selective group of patients and the fact that there was a high rate of young patients in this group (21.4% of them were under the age of 15). It was observed that there was a significant correlation between age and functional sensibility improvement (P = 0.04). Moreover, when we observed the relation between age and Semmes-Weinstein monofilament, static two-point discrimination, and dynamic two-point discrimination tests, it was seen that age had a significant correlation with each of the parameters (r = 0.61, P = 0.001; r = 0.58, P = 0.002; r = 0.57, P = 0.002). There was a clear decline in the intensity of the paraesthesia in S3+ and S4 group (X2 = 4.7, P = 0.02) and in these groups the period of Moberg pick-up test was meaningfully short (P < 0.05). PMID- 9727935 TI - Proton spectroscopy without water suppression: the oversampled J-resolved experiment. AB - A method is introduced for obtaining proton spectra in vivo with all the advantages of a full water signal. The method, based on F1 oversampled J-resolved spectroscopy, makes it possible to separate metabolite signals from unwanted baseline artifacts. The dominant water resonance is used as a 2D reference signal for the phase-sensitive reconstruction of the 2D J-resolved metabolite spectra. The powerful specificity of this method is demonstrated with model compound spectra, phantoms, and in vivo examples. PMID- 9727936 TI - A theoretical and experimental comparison of continuous and pulsed arterial spin labeling techniques for quantitative perfusion imaging. AB - Under ideal conditions, continuous arterial spin labeling (ASL) techniques are higher in SNR than pulsed ASL techniques by a factor of e. Presented here is a direct theoretical and experimental comparison of continuous ASL and pulsed ASL, using versions of both that are amenable to multislice imaging and insensitive to variations in transit times (continuous ASL with a delay before imaging, and QUIPSS II (Quantitative Imaging of Perfusion Using a Single Subtraction-second version)). Perfusion image quality for comparable imaging time was nearly identical for both single-slice and multislice imaging. The measured raw signal was approximately 25% higher with continuous ASL, but the SNR per unit time was identical. PMID- 9727937 TI - A rapid 2D time-resolved variable-rate k-space sampling MR technique for passive catheter tracking during endovascular procedures. AB - A new, fast, 2D MR imaging technique allowing passive catheter visualization adequate for use as a tool for guiding the movement of a catheter during endovascular procedures is described. This imaging technique samples low spatial frequencies more often than high spatial frequencies; it also uses both k-space view sharing and temporal interpolation. Unlike other techniques for passive visualization that exploit magnetic-susceptibility-induced artifacts, we have adopted a strategy that takes advantage of the T1-shortening effect of paramagnetic contrast agents, such as Gd-DTPA and a projection dephaser. This not only permits visualization of the entire catheter length but also minimizes the risk of intravascular heating. Using this method, a temporal frame rate of up to eight images per second and a tip localization accuracy of +/- 1mm (root mean square difference) can be achieved. PMID- 9727938 TI - Quantitative spectroscopic imaging of the human brain. AB - A method to provide B1 correction and cerebrospinal fluid (CSF) referencing is developed and applied to spectroscopic imaging of the human brain at 4.1 T using a volume head coil. The B1 image allows rapid determination of the spatially dependent B1 that is then used to compensate for the B1 sensitivity of the spectroscopic sequence. The reference signal is acquired from CSF located in a lateral ventricular position using a point-resolved echo spectroscopy (PRESS) acquisition. The CSF spectrum is also corrected for B1 dependence. Together with T2 and T1 corrections, this method is used to provide quantitative values of N acetylaspartate (NAA), creatine (Cr), and choline (Ch). The metabolite concentrations obtained from a spectroscopic imaging slice through the ventricles in seven normal controls are in good agreement with previously published literature values. This method is applied in a patient with secondary progressive multiple sclerosis, showing separate areas of abnormalities in both NAA and Cr. PMID- 9727939 TI - Sodium multiple quantum spectroscopy of articular cartilage: effects of mechanical compression. AB - The effects of mechanical compression on the multiple quantum coherences generated from sodium ions in articular cartilage were investigated. Cartilage samples obtained from bovine patellae were studied during compression at 0.7 MPa (100 psi) for 1 hour. The double quantum filtered spectra showed marked lineshape changes in the compressed samples. Compression did not seem to influence the lineshapes of the single quantum and triple quantum filtered spectra significantly. We found that the residual quadrupolar interaction was reduced in the compressed samples. Changes in the ordering of collagen fibers may be responsible for the observed effect. PMID- 9727940 TI - Quantitative assessment of improved homogeneity using higher-order shims for spectroscopic imaging of the brain. AB - Magnetic field homogeneity is of major concern for in vivo spectroscopy, and with the increased use of volumetric chemical shift imaging (CSI) techniques, the ability to shim over a large volume of tissue is now one of the primary limiting constraints in performing these studies. In vivo shimming is routinely performed using linear shim correction terms, and although many scanners are also equipped with additional resistive shim supplies that can provide second and third-order shim fields, they are often not used due to the additional effort and scan time required. The question as to how much improvement can be achieved using additional higher-order shims compared with the linear shims alone was quantitatively addressed. Performance measures for both intervoxel B0 uniformity and intravoxel T2* line broadening were evaluated for 15 normal volunteers. The analysis tools developed in this study, along with the summarized data, can be useful in deciding if a given application warrants the additional time, effort, and expense (if additional hardware needs to be purchased) of implementing higher order shimming routines. For CSI studies of the brain, the use of the higher order shims, compared with linear terms alone, yielded approximately 30% greater volume of brain tissue that could be shimmed within typical constraints for intervoxel B0 shifts and intravoxel T2* linebroadening. In addition, a regional analysis shows significant improvement in the homogeneity within specific areas of the brain, particularly those near the skull. PMID- 9727941 TI - A general kinetic model for quantitative perfusion imaging with arterial spin labeling. AB - Recently, several implementations of arterial spin labeling (ASL) techniques have been developed for producing MRI images sensitive to local tissue perfusion. For quantitation of perfusion, both pulsed and continuous labeling methods potentially suffer from a number of systematic errors. In this study, a general kinetic model for the ASL signal is described that can be used to assess these errors. With appropriate assumptions, the general model reduces to models that have been used previously to analyze ASL data, but the general model also provides a way to analyze the errors that result if these assumptions are not accurate. The model was used for an initial assessment of systematic errors due to the effects of variable transit delays from the tagging band to the imaging voxel, the effects of capillary/tissue exchange of water on the relaxation of the tag, and the effects of incomplete water extraction. In preliminary experiments with a human subject, the model provided a good description of pulsed ASL data during a simple sensorimotor activation task. PMID- 9727942 TI - Measurement of tumor vascular volume and mean microvascular random flow velocity magnitude by dynamic Gd-DTPA-albumin enhanced and diffusion-weighted MRI. AB - Tumor vascular volume fraction and the magnitude of the mean microvascular random flow velocity were measured in an animal tumor model by combining dynamic Gd-DTPA albumin enhanced MRI and diffusion-weighted MRI in conjunction with a compartmental modeling analysis. The vascular volume fraction maps were obtained from the dynamic Gd-DTPA-albumin enhanced MRI measurement. It was found that the vascular volume fraction for Walker 256 tumor was higher within the outgrowing rim and decreased towards the central region. The average value obtained from five animals was 0.062 +/- 0.009 ml/g. By using the vascular volume fraction from the Gd-DTPA-albumin enhanced MRI measurement, maps of the magnitude of the mean microvascular random flow velocity were obtained from the diffusion-weighted MRI measurements with the compartmental modeling analysis. The relative extravascular and intravascular contributions to the diffusion-weighted MRI signal were determined for three tissue groups with different Gd-DTPA-albumin enhancement characteristics, and the flow and molecular diffusion-induced attenuation factors for the intravascular compartment were also compared. The mean microvascular random flow velocity magnitude maps were obtained with an average value of 0.67 +/- 0.06 mm/s. PMID- 9727943 TI - Diffusional anisotropy of T2 components in bovine optic nerve. AB - A diffusion-CPMG hybrid experiment was used to analyze the diffusion characteristics of different T2 relaxation components in bovine optic nerve. Data were collected using a pulsed field gradient (PFG) multi spin echo (MSE) CPMG sequence for parallel and perpendicular axon orientation and four diffusion times. The apparent diffusion coefficient (ADC) was evaluated for two observed T2 components as a function of axonal orientation and diffusion time delta. The short T2 component exhibited minor diffusional anisotropy and larger ADC, whereas the long T2 component showed significant anisotropy effects. This is consistent with the hypothesis that the short T2 component is associated with water within the myelin sheath, which is less restricted than axonal water that is limited by cell membrane permeability. PMID- 9727945 TI - Functional MRI of the rat somatosensory cortex: effects of hyperventilation. AB - Functional mapping of the rat somatosensory cortex was performed with T2* sensitized MRI using a forepaw electrical stimulation model in alpha-chloralose anesthetized rats at 7 T under both normocapnia and mild hyperventilation-induced hypocapnia. A highly localized activation area, consistent with the known somatosensory cortical region, was detected in all seven animals studied during hypocapnia and in five of the same animals during normocapnia. Quantitatively, hypocapnia was found to significantly increase both the size of the fMRI activation area (3.4 +/- 0.6 mm2 versus 1.5 +/- 0.6 mm2 in normocapnia, mean +/- standard error, n = 7, P < 0.03) and the average fMRI signal intensity increase (3.4 +/- 0.6% versus 2.7 +/- 0.4%, n = 5, P < 0.05). The increased sensitivity of fMRI to functional activation may reflect a widened arterial-venous oxygenation difference resulting from an increased effective oxygen extraction during hyperventilation. The dependence of the fMRI response on the ventilation state underscores the need to control for physiological parameters in animal fMRI studies. PMID- 9727944 TI - Induction of apoptosis in two mammalian cell lines results in increased levels of fructose-1,6-bisphosphate and CDP-choline as determined by 31P MRS. AB - Programmed cell death or apoptosis was induced in human promyelocytic leukemia (HL-60) and Chinese hamster ovary (CHO-K1) cells using several cytotoxic drugs that have different modes of action, including camptothecin, ceramide, chelerythrine, etoposide, farnesol, geranyl geraniol, and hexadecylphosphocholine. The consequent changes in cellular metabolism were monitored using 31P MRS measurements on intact cells and cell extracts. Cells undergoing programmed cell death exhibited characteristic changes in the levels of glycolytic and phospholipid metabolites. The most significant changes were increases in the concentration of the glycolytic intermediate, fructose-1,6 bisphosphate and in the concentration of CDP-choline, which is an intermediate in phosphatidylcholine biosynthesis. In HL-60 cells, the increase in fructose-1,6 bisphosphate levels could be explained by depletion of cellular NAD(H) levels. All of the agents used to induce apoptosis caused the accumulation of CDP choline. Since the resonances of this compound occur in a relatively well resolved region of tissue spectra, it could provide a marker for apoptosis that would allow the noninvasive detection of the process in vivo using 31P MRS measurements. PMID- 9727946 TI - A programmable pre-emphasis system. AB - MRI systems often use magnetic field gradient and shim pulse-shaping networks (pre-emphasis) to correct for magnetic field distortions caused by eddy currents. A pre-emphasis system that uses up to 16 fixed resistor-capacitor (RC) time constants per channel with programmable amplitude coefficients is described. The magnetic fields induced by the pre-emphasis RC time constants serve as a set of basis functions for compensating eddy-current fields induced by the gradient set. The resultant time-varying magnetic field gradient accurately reflects the gradient specified by the pulse programmer. Reductions in eddy-current fields are demonstrated for actively shielded and unshielded gradient sets. PMID- 9727947 TI - Asymmetric spin-echo imaging of magnetically inhomogeneous systems: theory, experiment, and numerical studies. AB - The ability of the asymmetric spin-echo (ASE) pulse sequence to provide different degrees of spin-echo (SE)-type and gradient-echo (GE)-type contrast when imaging media containing magnetic inhomogeneities is investigated. The dependence of the ASE signal on the size of magnetic field perturbers is examined using theory, computer simulations, and experiment. A theoretical prediction of the ASE signal is obtained using the Anderson-Weiss mean field theory, the results of which are qualitatively supported by computer simulations and experimental studies. It is shown that the ASE sequence can be used to tune the range of perturber sizes that provide the largest contributions to susceptibility contrast effects. PMID- 9727948 TI - Feature space analysis of MRI. AB - This paper presents an MRI feature-space image-analysis method and its application to brain tumor studies. The proposed method generates a transformed feature space in which the normal tissues (white matter, gray matter, and CSF) become orthonormal. As such, the method is expected to have site-to-site and patient-to-patient consistency, and is useful for identification of tissue types, segmentation of tissues, and quantitative measurements on tissues. The steps of the work accomplished are as follows: (1) Four T2-weighted and two T1-weighted images (before and after injection of gadolinium) were acquired for 10 tumor patients. (2) Images were analyzed by an image analyst according to the proposed algorithm. (3) Biopsy samples were extracted from each patient and were subsequently analyzed by the pathology laboratory. (4) Image-analysis results were compared with the biopsy results. Pre- and postsurgery feature spaces were also compared. The proposed method made it possible to visualize the MRI feature space and to segment the image. In all cases, the operators were able to find clusters for normal and abnormal tissues. Also, clusters for different zones of the tumor were found. The method successfully segmented the image into normal tissues (white matter, gray matter, and CSF) and different zones of the lesion (tumor, cyst, edema, radiation necrosis, necrotic core, and infiltrated tumor). The results agreed with those obtained from the biopsy samples. Comparison of pre with postsurgery and radiation feature spaces illustrated that the original solid tumor was not present in the second study, but a new tissue component appeared in a different location of the feature space. This tissue could be radiation necrosis generated as a result of radiation. PMID- 9727949 TI - Ex vivo tissue-type independence in proton-resonance frequency shift MR thermometry. AB - The temperature sensitivity of the proton-resonance frequency (PRF) has proven valuable for the monitoring of MR image-guided thermal coagulation therapy. However, there is significant inconsistency in reported values of the PRF-thermal coefficient, as measured from experiments encompassing a range of in vivo and ex vivo tissue types and experimental conditions. A method of calibrating the temperature dependence of the PRF is described and results are presented that indicate a tissue-type independence. To this end, other possible mechanisms for variations in the PRF-thermal coefficient are suggested, including physiological perturbations and volume magnetic susceptibility effects from geometry and orientation. PMID- 9727950 TI - Sampling errors in projection reconstruction MRI. AB - Certain kinds of artifacts have been reported when projection reconstruction (PR) techniques are used in magnetic resonance imaging (MRI). These will occur if the spacing between samples in k-space is too large. It has been suggested that PR requires finer k-space sampling than does two-dimensional Fourier reconstruction, and that the usual Nyquist criterion is inadequate. This paper examines this problem, with the conclusion that the Nyquist sampling criterion is adequate to avoid aliasing effects, provided that a band-limited interpolation is used in k space. This procedure is motivated by analysis of the PR technique as it is commonly implemented in x-ray computed tomography. A related problem is shown to be the construction of a filter function in k-space that gives proper weight to the low spatial frequencies. It is shown that a simple ?k? filter does not satisfy this requirement, and a procedure for deriving a suitable filter is described. The methods are tested in simulated PR profiles of circular disks of two different sizes. It is shown that the combination of the two new methods gives virtually perfect reconstruction for disks up to the size implied by the Nyquist limit. PMID- 9727951 TI - In vivo perfusion measurements in the human placenta using echo planar imaging at 0.5 T. AB - This paper presents the first in vivo measurements of perfusion in the human placenta from 20 weeks gestational age until term, using the non selective/selective inversion recovery echo-planar imaging sequence, in which data is alternately acquired following a selective and non-selective inversion pulse. Twenty pairs of images were collected, two each at the following inversion times: 20, 310, 610, 910, 1110, 1410, 1910, 2810, 3310, and 4510 ms with the sequence being repeated with a repetition time (TR) of 10 s. The results of these measurements were used to suggest the optimum sequence for future work in terms of the signal to noise ratio in the measured perfusion rate in a given measurement time. The sequence was also analyzed to determine the expected variability in the measurements. In normal pregnancies the average value of perfusion rate was found to be 176 (standard error = +/-24) ml/100 mg/min. (n = 16, standard deviation = 96 ml/100 mg/min). The expected variability in the measured parameters due to signal to noise ratio considerations alone was calculated to be 71%. For a maximum scanning time of 400 s, the optimum sequence for measuring placental perfusion was found to require 8 repetitions at each of 10 inversion times which were geometrically spaced (given by a(o), a(o)r, a(o)r2, a(o)r3, . . .), with a(o) = 850 ms, r = 1.073 and TR = 5 s, giving a pixel variability of 38%. Other timing schemes are recommended for measuring perfusion in other anatomical regions with different values of perfusion rate and longitudinal relaxation time. PMID- 9727952 TI - Reduced circular field-of-view imaging. AB - A rapid dynamic imaging technique based on polar k-space sampling is presented. A gain in temporal resolution is achieved by angular undersampling. A detailed analysis of the point spread function of angular undersampled polar imaging reveals a reduced diameter of the corresponding circular field of view. Under the assumption that dynamic changes are restricted to a local circular field of view, angular undersampled dynamic imaging allows the recording of rapid changes at high temporal and spatial resolution. The theoretical and experimental details of the technique are presented. PMID- 9727953 TI - Partial Fourier reconstruction for three-dimensional gradient echo functional MRI: comparison of phase correction methods. AB - Partial Fourier (PF) methods take advantage of data symmetry to allow for either faster image acquisition or increased image resolution. Faster acquisition and increased spatial resolution are advantageous for fMRI because of increased temporal resolution and/or reduced partial volume effects, respectively. Standard PF methods, which use a phase reference obtained from a low resolution image, are adequate for the reconstruction of time-stationary images acquired using either spin echoes or short TE gradient echoes. In fMRI, however, multiple images are acquired using long TE gradient echoes, which introduces possible phase drifts in the fMRI data and high spatial frequencies in the phase reference. This work investigates several techniques developed to reconstruct fMRI data obtained with PF acquisitions. PF methods that account for both high-frequency spatial variations and time-dependent drifts in the phase reference are discussed and are quantitatively evaluated using receiver operator characteristic curve analysis. PMID- 9727954 TI - ESR spectroscopy for analysis of hippocampal elimination of a nitroxide radical during kainic acid-induced seizure in rats. AB - To analyze the balance between free radical production and elimination during seizure, techniques were developed to monitor the sequential changes in the level of an exogenous free radical by ESR in the brains of freely moving rats given kainic acid (KA) to induce seizures (n = 6) and control rats given saline (n = 6). The hippocampus of each rat was perfused with a nitroxide radical solution for 120 min by in vivo microdialysis. Then an intraperitoneal injection of KA or the physiological saline was given to each rat, and the perfusate was changed to Ringer's solution. ESR analysis of sequential samples of dialysate taken after injection revealed an exponential decay in signal amplitude. The median half-life of the nitroxide radical was significantly longer in the KA-treated group than in the control group (P < 0.01). This finding demonstrates a decreased capacity of the hippocampus to eliminate the nitroxide radical from the extracellular space during seizure and may help clarify the mechanism of neuronal cell vulnerability to free radicals during insult or degeneration. PMID- 9727955 TI - Partial volume effects on vascular T2 measurements. AB - Quantitative, vascular T2 measurements are of interest for applications such as MR oximetry. In the situation of a vessel with long T2 relaxation times embedded in tissue with relatively short T2 values, contamination of the blood signal from the surrounding tissue can bias T2 measurements. Limited data sampling and vessels running obliquely through the imaging slice can cause significant signal contamination. Using a model of these effects to predict the behavior of T2 measurements under a range of conditions, a set of parameters that provides the best combination of measurement accuracy with a signal-to-noise ratio as high as possible is proposed. PMID- 9727956 TI - Optimal design of gradient coils in MR imaging: optimizing coil performance versus minimizing cost functions. AB - The "forward" method of an optimal gradient coil design provides a coil that has the minimal cost function value. It is shown in this study that the solution obtained by minimizing the cost function is directly dependent on the specified cost function and generally results in a deviation from the most desirable coil design. In this paper, a gradient coil design approach for obtaining the best achievable coil performance for pre-determined imaging applications is presented. Through this approach, all intermediate coil performance values calculated during an optimization process, using a simulated annealing algorithm, are stored and presented in a three-dimensional data set. Using this three-dimensional data set, a coil designer is able to make a balance between different coil performance parameters and to select a coil that is the most desirable for the pre-determined imaging applications. PMID- 9727957 TI - Preventing and controlling oral and pharyngeal cancer. Recommendations from a National Strategic Planning Conference. AB - In August 1996, CDC convened a national conference to develop strategies for preventing and controlling oral and pharyngeal cancer in the United States. The conference, which was cosponsored by the National Institute of Dental Research of the National Institutes of Health and the American Dental Association, included 125 experts in oral and pharyngeal cancer prevention, treatment, and research; both the private and public sectors were represented. Participants at the conference developed recommendations concerning advocacy, collaboration, and coalition building; public health policy; public education; professional education and practice; and data collection, evaluation, and research. A follow up meeting consisting of selected participants of the 1996 conference was held in September 1997. During this meeting, changes that had occurred in the political and scientific arenas since the 1996 conference were considered, and 10 recommended strategies from the conference were selected for priority implementation. These 10 strategies were to a) establish a mechanism to implement and monitor the recommended strategies developed during the conference; b) urge oral health professionals to become more actively involved in community health; c) require instruction in preventing and controlling tobacco and alcohol use at all levels of training in dental, medical, nursing, and other related health-care disciplines; d) encourage Medicaid, Medicare, traditional insurance plans, and managed-care entities to consider making oral cancer examinations an integral part of comprehensive physical and oral examinations; e) designate federal funding for a national program of oral cancer prevention, early detection, and control; f) after assessing local needs, develop, implement, and evaluate statewide models to educate all relevant groups; g) develop and conduct a national promotional campaign to raise public awareness of oral cancer and its link to tobacco use and heavy alcohol consumption; h) develop health-care curricula that require competency in prevention, diagnosis, and multidisciplinary management of oral and pharyngeal cancer; i) sponsor and promote continuing education for health-care professionals on the multidisciplinary management of all phases of oral cancer and its sequelae; and j) strengthen organizational approaches to reducing oral cancer by developing organized cooperative and collaborative arrangements, funding formal centers, and involving commercial firms. CDC will use these recommended strategies to develop programs to reduce the burden of oral and pharyngeal cancer in the United States. Through the Oral Cancer Roundtable, a group of conference and meeting participants, CDC will communicate to interested agencies, organizations, and state health departments ways in which they can implement elements of the national plan. The Roundtable will help CDC track the efforts and progress of these groups. PMID- 9727958 TI - Development of new vaccines for tuberculosis. Recommendations of the Advisory Council for the Elimination of Tuberculosis (ACET). AB - Tuberculosis (TB) remains a major, global public health problem, particularly in low-income countries. Better application of current diagnostic, treatment, and prevention strategies could lead to gradual decreases in the disease, but eliminating TB completely in the United States and internationally will require new tools. The greatest impact could come from a new vaccine, and recent technological advances have provided the basis for new vaccine development. However, sustained support is required to move the research from the laboratory to field trials of vaccines and to implement new vaccine programs. Recognizing the importance of TB vaccines, the Advisory Council for the Elimination of Tuberculosis (ACET) recommends that public agencies and vaccine manufacturers develop a comprehensive, consensual strategy to achieve these goals. This report outlines the elements that should be considered in devising a strategic plan for vaccine development. PMID- 9727959 TI - Hostility is associated with a heightened prolactin response to meta chlorophenylpiperazine in abstinent cocaine addicts. AB - The prolactin (PRL) response to the administration of serotonin (5HT) agonists is an index of central nervous system 5HT activity. This index is blunted in association with hostile aggression in personality and depressive disorder patients without substance abuse. We tested whether the PRL response to the oral administration of the partial 5HT agonist meta-chlorophenylpiperazine (MCPP), 0.35 mg/kg, was associated with a measure of trait hostility, the Buss Durkee Hostility Inventory (BDHI), in cocaine addicts who were completing a 3-week detoxification and rehabilitation program. We also tested whether the cocaine addicts differed from healthy volunteers on their PRL, cortisol (CORT) or temperature responses to MCPP. The PRL response to MCPP was positively associated with the total score on the BDHI. There were, however, no differences in the neuroendocrine or temperature responses to MCPP between the cocaine-dependent group and the healthy volunteers once age effects were controlled for. PMID- 9727960 TI - Blunted cardiovascular and catecholamine stress reactivity in women with bulimia nervosa. AB - Cardiovascular and catecholamine responses to mental stressors were investigated in women with bulimia nervosa (BN) and in healthy control subjects. Fifteen women with BN and 15 control subjects completed psychosocial questionnaires before laboratory testing, where they were exposed to an interpersonally based speech stressor and a serial math task. Blood pressure, heart rate, epinephrine, norepinephrine and, via impedance cardiography, systolic time intervals, cardiac output and total peripheral resistance were measured at rest and during stress. Results indicated that BN was associated with blunted sympathetic activation in response to mental stress, indicated by increased pre-ejection period responses and blunted systolic blood pressure, heart rate and epinephrine responses. In contrast, women with BN had elevated cortisol levels when compared with control women. In addition, despite equivalent performance between groups, bulimic women reported feeling significantly more confused, frustrated, inadequate and dissatisfied with their performance during tasks. Psychosocial questionnaires also indicated that women with BN perceived more stress, had worse coping skills, lower self-esteem and sense of mastery, reported less social support, had worse mood, had greater anxiety and were more depressed when compared with control women. These results are interpreted as reflecting physiological and psychological profiles indicative of distress vs. active effort coping in BN. PMID- 9727961 TI - Reduced level of plasma antioxidant uric acid in schizophrenia. AB - There is evidence of dysregulation of the antioxidant defense system in schizophrenia. The purpose of the present study was to examine whether uric acid, a potent antioxidant, is reduced in the plasma of patients with schizophrenia. To this end, a within-subject, repeated measures, on-off-on haloperidol treatment design was utilized. Male schizophrenic patients with either a haloperidol treatment (n=47) or a drug-free condition (n=35) had significantly lower levels of plasma uric acid than the age- and sex-matched normal control subjects (n=34). Following haloperidol withdrawal, plasma uric acid levels were further reduced in schizophrenic patients (P=0.018; paired t-test, n=35). However, no relationship was found between uric acid levels and the length of the drug-free period (< 5 or > 5 weeks) or days drug free. In addition, the plasma levels of uric acid in patient groups were significantly and inversely correlated with psychosis. There was a trend for lower uric acid levels in relapsed patients relative to clinically stable patients. Smoking, which can modify plasma antioxidant capacity, was not found to have prominent effects on uric acid levels. The present finding of a significant decrease of a selective antioxidant provides additional support to the hypothesis that oxidative stress in schizophrenia may be due to a defect in the antioxidant defense system. PMID- 9727962 TI - Correlates of functional status in older patients with schizophrenia. AB - There is a growing recognition of the importance of quantifying the impact of illness on functional abilities. Measures of function frequently rely on a self report. Few studies have directly assessed functional capacity in psychiatric patients, especially older ones who may be at an increased risk for disability. Subjects were 102 middle-aged and elderly outpatients with DSM-III-R or DSM-IV diagnosis of schizophrenia or schizoaffective disorder, and 66 normal comparison subjects, ranging in age from 45 to 86. The Direct Assessment of Functional Status (DAFS), a standardized measure of behavior during simulated daily activity tasks (i.e. time orientation, communication, transportation, finance, shopping, grooming and eating) was used to quantify levels of disability. Schizophrenic patients demonstrated significantly greater disability than normal subjects. An evaluation of specific behaviors indicated that the patients were significantly more limited than comparison subjects across all subscales of the DAFS except for grooming and eating. A lower level of formal education, greater severity of extrapyramidal symptoms, and greater cognitive deficits, but not severity of symptoms of schizophrenia, were related to lower DAFS scores. Relative to published findings, schizophrenic patients appeared more disabled than outpatients with major depression, but less disabled than those with Alzheimer's disease. The DAFS is a useful instrument for characterizing functional abilities in older patients with schizophrenia. Our findings of significant functional disability in older schizophrenic patients have implications for treatment as well as allocation of health-care resources. PMID- 9727963 TI - Prediction of the deficit syndrome from initial deficit symptoms in the early course of schizophrenia. AB - The Proxy for the Deficit Syndrome (PDS) was used with longitudinal symptom assessment data to identify recent-onset schizophrenia patients with the deficit syndrome. We evaluated the stability of deficit symptoms using repeated assessments. Symptom ratings were examined at an initial point of outpatient stabilization on antipsychotic medication as well as prospectively over the subsequent 12 months of outpatient treatment and assessment in 83 recent-onset schizophrenia patients. The vast majority of patients who were classified as non deficit at the cross-sectional baseline assessment continued to remain non deficit throughout the first year of treatment. However, patients classified as deficit at baseline did not consistently remain classified as showing deficit syndrome during the follow-through period. Thus, the presence of deficit symptoms detected in a single cross-sectional rating may be an inaccurate way to rate the deficit syndrome, yielding excessive false positives. Our use of longitudinal data allowed the stability criterion of the deficit syndrome to be evaluated using the PDS. PMID- 9727964 TI - Personality and psychosocial dysfunction in schizophrenia: the association of extraversion and neuroticism to deficits in work performance. AB - Research on vocational dysfunction in schizophrenia has as yet only examined associated features of illness. We hypothesized that personality variables may be also associated with work function. We reasoned that higher levels of extraversion and neuroticism would predict poor function by virtue of the social support seeking and passive/avoidant coping styles associated with each. To test this, multiple regressions were conducted in which measures of extraversion and neuroticism predicted work performance among 43 subjects with schizophrenia or schizoaffective disorder. Higher levels of extraversion and neuroticism significantly predicted poorer function, accounting for between 7% and 27% of the variance in global cooperativeness, work quality, work habits and personal presentation measures of work behavior. The potential importance of assessing personality in rehabilitation is discussed. PMID- 9727965 TI - Illusory conjunctions and perceptual grouping in a visual search task in schizophrenia. AB - This report describes part of a series of experiments, conducted within the framework of feature integration theory, to determine whether patients with schizophrenia show deficits in preattentive processing. Thirty subjects with a DSM-III-R diagnosis of schizophrenia and 30 age-, gender-, and education-matched normal control subjects completed two computerized experimental tasks, a visual search task assessing the frequency of illusory conjunctions (i.e. false perceptions) under conditions of divided attention (Experiment 3) and a task which examined the effects of perceptual grouping on illusory conjunctions (Experiment 4). We also assessed current symptomatology and its relationship to task performance. Contrary to our hypotheses, schizophrenia subjects did not show higher rates of illusory conjunctions, and the influence of perceptual grouping on the frequency of illusory conjunctions was similar for schizophrenia and control subjects. Nonetheless, specific predictions from feature integration theory about the impact of different target types (Experiment 3) and perceptual groups (Experiment 4) on the likelihood of forming an illusory conjunction were strongly supported, thereby confirming the integrity of the experimental procedures. Overall, these studies revealed no firm evidence that schizophrenia is associated with a preattentive abnormality in visual search using stimuli that differ on the basis of physical characteristics. PMID- 9727966 TI - Auditory and visual working memory performance in patients with frontal lobe damage and in schizophrenic patients with low scores on the Wisconsin Card Sorting Test. AB - Impaired performance in the Wisconsin Card Sorting Test (WCST) has frequently been postulated to be one typical feature indicating frontal dysfunction in patients with schizophrenia. From a functional point of view, impairments were attributed to a dysfunction of working memory. The present study compares the performance of groups of schizophrenic patients, groups of patients with acquired brain damage as well as normal controls on tasks involving visual and auditory working memory. Modified versions of Sternberg tasks were used varying the physical attributes of the material to be rehearsed in order to force a different involvement of both the 'visuo-spatial sketch-pad' and the 'phonological loop'. Within a group of frontal-lobe-damaged patients (n=6), processing was markedly prolonged for both kinds of material, an observation attributed to a dysfunction of the central executive component of the working memory model. On the other hand, results for schizophrenic patients with poor WCST performance (n=6) suggest a more discrete dysfunction of the phonological loop, but not the visuo-spatial sketch-pad. There were no significant differences between normal controls (n=6) and clinical control groups [patients with non-frontal lesions (n=6) and schizophrenic patients with normal scores on the WCST (n=6)]. Comparisons of the various group data rule out an unspecified frontal dysfunction of schizophrenic patients with low scores on the WCST. PMID- 9727967 TI - Lunar Prospector: overview. AB - Lunar Prospector is providing a global map of the composition of the moon and analyzing the moon's gravity and magnetic fields. It has been in a polar orbit around the moon since 16 January 1998. Neutron flux data show that there is abundant H, and hence probably abundant water ice, in the lunar polar regions. Gamma-ray and neutron data reveal the distribution of Fe, Ti, and other major and trace elements on the moon. The data delineate the global distributions of a key trace element-rich component of lunar materials called KREEP and of the major rock types. Magnetic mapping shows that the lunar magnetic fields are strong antipodal to Mare Imbrium and Mare Serenitatis and has discovered the smallest known magnetosphere, magnetosheath, and bow shock complex in the solar system. Gravity mapping has delineated seven new gravity anomalies and shown that the moon has a small Fe-rich core of about 300 km radius. PMID- 9727968 TI - Improved gravity field of the moon from lunar prospector AB - An improved gravity model from Doppler tracking of the Lunar Prospector (LP) spacecraft reveals three new large mass concentrations (mascons) on the nearside of the moon beneath the impact basins Mare Humboltianum, Mendel-Ryberg, and Schiller-Zucchius, where the latter basin has no visible mare fill. Although there is no direct measurement of the lunar farside gravity, LP partially resolves four mascons in the large farside basins of Hertzsprung, Coulomb-Sarton, Freundlich-Sharonov, and Mare Moscoviense. The center of each of these basins contains a gravity maximum relative to the surrounding basin. The improved normalized polar moment of inertia (0.3932 +/- 0.0002) is consistent with an iron core with a radius of 220 to 450 kilometers. PMID- 9727969 TI - Lunar surface magnetic fields and their interaction with the solar wind: results from lunar prospector AB - The magnetometer and electron reflectometer experiment on the Lunar Prospector spacecraft has obtained maps of lunar crustal magnetic fields and observed the interaction between the solar wind and regions of strong crustal magnetic fields at high selenographic latitude (30 degreesS to 80 degreesS) and low ( approximately 100 kilometers) altitude. Electron reflection maps of the regions antipodal to the Imbrium and Serenitatis impact basins, extending to 80 degreesS latitude, show that crustal magnetic fields fill most of the antipodal zones of those basins. This finding provides further evidence for the hypothesis that basin-forming impacts result in magnetization of the lunar crust at their antipodes. The crustal magnetic fields of the Imbrium antipode region are strong enough to deflect the solar wind and form a miniature (100 to several hundred kilometers across) magnetosphere, magnetosheath, and bow shock system. PMID- 9727971 TI - Major compositional units of the moon: lunar prospector thermal and fast neutrons AB - Global maps of thermal and fast neutron fluxes from the moon suggest three end member compositional units. A high thermal and low fast neutron flux unit correlates with the lunar highlands and is consistent with feldspathic rocks. The South Pole-Aitken basin and a strip that surrounds the nearside maria have intermediate thermal and fast neutron flux levels, consistent with more mafic rocks. There appears to be a smooth transition between the most mafic and feldspathic compositions, which correspond to low and high surface altitudes, respectively. The maria show low thermal and high fast neutron fluxes, consistent with basaltic rocks. PMID- 9727970 TI - Global elemental maps of the moon: the Lunar Prospector gamma-Ray spectrometer. AB - Lunar Prospector gamma-ray spectrometer spectra along with counting rate maps of thorium, potassium, and iron delineate large compositional variations over the lunar surface. Thorium and potassium are highly concentrated in and around the nearside western maria and less so in the South Pole-Aitken basin. Counting rate maps of iron gamma-rays show a surface iron distribution that is in general agreement with other measurements from Clementine and the Lunar Prospector neutron detectors. PMID- 9727972 TI - Lunar Fe and Ti abundances: comparison of lunar prospector and clementine data AB - The Lunar Prospector neutron spectrometer data correlate well with iron and titanium abundances obtained through analysis of Clementine spectral reflectance data. With the iron and titanium dependence removed, the neutron spectrometer data also reveal regions with enhanced amounts of gadolinium and samarium, incompatible rare earth elements that are enriched in the final phases of magma crystallization. These regions are found mainly around the ramparts of the Imbrium impact basin but not around the other basins, including the much larger and deeper South Pole-Aitken basin. This result confirms the compositional uniqueness of the surface and interior of the Imbrium region. PMID- 9727973 TI - Fluxes of fast and epithermal neutrons from Lunar Prospector: evidence for water ice at the lunar poles. AB - Maps of epithermal- and fast-neutron fluxes measured by Lunar Prospector were used to search for deposits enriched in hydrogen at both lunar poles. Depressions in epithermal fluxes were observed close to permanently shaded areas at both poles. The peak depression at the North Pole is 4.6 percent below the average epithermal flux intensity at lower latitudes, and that at the South Pole is 3.0 percent below the low-latitude average. No measurable depression in fast neutrons is seen at either pole. These data are consistent with deposits of hydrogen in the form of water ice that are covered by as much as 40 centimeters of desiccated regolith within permanently shaded craters near both poles. PMID- 9727974 TI - Low-temperature synthesis of zintl compounds with a single-source molecular precursor AB - Thermolysis of the heterobimetallic phosphinidene complex [Sb(PCy)3]2- Li6.6HNMe2 (Cy = C6H11) at 303 to 313 kelvin gives Zintl compounds containing (Sb7)3- anions. The complex thus constitutes a stable molecular single-source precursor to Zintl compounds and provides a potential low-temperature route to photoactive alkali metal antimonates. The new chemical reaction involved, which is driven thermodynamically by the formation of P-P bonds, has implications in the low temperature synthesis of other technologically important materials (such as gallium arsenide). PMID- 9727975 TI - Segregation of transcription and replication sites into higher order domains. AB - Microscopy shows that individual sites of DNA replication and transcription of mammalian nuclei segregate into sets of roughly 22 and 16 higher order domains, respectively. Each domain set displayed a distinct network-like appearance, including regions of individual domains and interdigitation of domains between the two networks. These data support a dynamic mosaic model for the higher order arrangement of genomic function inside the cell nuclei. PMID- 9727976 TI - CBP: a signal-regulated transcriptional coactivator controlled by nuclear calcium and CaM kinase IV. AB - Recruitment of the coactivator, CREB binding protein (CBP), by signal-regulated transcription factors, such as CREB [adenosine 3', 5'-monophosphate (cAMP) response element binding protein], is critical for stimulation of gene expression. The mouse pituitary cell line AtT20 was used to show that the CBP recruitment step (CREB phosphorylation on serine-133) can be uncoupled from CREB/CBP-activated transcription. CBP was found to contain a signal-regulated transcriptional activation domain that is controlled by nuclear calcium and calcium/calmodulin-dependent (CaM) protein kinase IV and by cAMP. Cytoplasmic calcium signals that stimulate the Ras mitogen-activated protein kinase signaling cascade or expression of the activated form of Ras provided the CBP recruitment signal but did not increase CBP activity and failed to activate CREB- and CBP mediated transcription. These results identify CBP as a signal-regulated transcriptional coactivator and define a regulatory role for nuclear calcium and cAMP in CBP-dependent gene expression. PMID- 9727977 TI - Identification of c-MYC as a target of the APC pathway. AB - The adenomatous polyposis coli gene (APC) is a tumor suppressor gene that is inactivated in most colorectal cancers. Mutations of APC cause aberrant accumulation of beta-catenin, which then binds T cell factor-4 (Tcf-4), causing increased transcriptional activation of unknown genes. Here, the c-MYC oncogene is identified as a target gene in this signaling pathway. Expression of c-MYC was shown to be repressed by wild-type APC and activated by beta-catenin, and these effects were mediated through Tcf-4 binding sites in the c-MYC promoter. These results provide a molecular framework for understanding the previously enigmatic overexpression of c-MYC in colorectal cancers. PMID- 9727978 TI - Extrapolating species abundance across spatial scales AB - The analysis, measurement, and management of species abundance is central to ecology and conservation biology, but it has proved difficult to find a single index that adequately reflects the commonness or rarity of species across a range of spatial scales. Here, a scale-independent measure of species abundance is developed, using presence-absence maps at varying spatial resolutions. By extrapolating such "scale-area" curves, species abundance can be estimated accurately even at scales finer than those used to parameterize the model, a task that had previously been deemed impossible in principle. PMID- 9727979 TI - Conversion of neuronal growth cone responses from repulsion to attraction by cyclic nucleotides. AB - Nerve growth is regulated by attractive and repulsive factors in the nervous system. Microscopic gradients of Collapsin-1/Semaphorin III/D (Sema III) and myelin-associated glycoprotein trigger repulsive turning responses by growth cones of cultured Xenopus spinal neurons; the repulsion can be converted to attraction by pharmacological activation of the guanosine 3',5'-monophosphate (cGMP) and adenosine 3',5'-monophosphate signaling pathways, respectively. Sema III also causes the collapse of cultured rat sensory growth cones, which can be inhibited by activation of the cGMP pathway. Thus cyclic nucleotides can regulate growth cone behaviors and may be targets for designing treatments to alleviate the inhibition of nerve regeneration by repulsive factors. PMID- 9727980 TI - Expression of a gene cluster kaiABC as a circadian feedback process in cyanobacteria. AB - Cyanobacteria are the simplest organisms known to have a circadian clock. A circadian clock gene cluster kaiABC was cloned from the cyanobacterium Synechococcus. Nineteen clock mutations were mapped to the three kai genes. Promoter activities upstream of the kaiA and kaiB genes showed circadian rhythms of expression, and both kaiA and kaiBC messenger RNAs displayed circadian cycling. Inactivation of any single kai gene abolished these rhythms and reduced kaiBC-promoter activity. Continuous kaiC overexpression repressed the kaiBC promoter, whereas kaiA overexpression enhanced it. Temporal kaiC overexpression reset the phase of the rhythms. Thus, a negative feedback control of kaiC expression by KaiC generates a circadian oscillation in cyanobacteria, and KaiA sustains the oscillation by enhancing kaiC expression. PMID- 9727982 TI - Antimicrobial resistance: a veterinary perspective. Antimicrobials are important for animal welfare but need to be used prudently. PMID- 9727981 TI - Antimicrobial resistance. Is a major threat to public health. PMID- 9727984 TI - Control of antimicrobial resistance: time for action. The essentials of control are already well known. PMID- 9727983 TI - Antimicrobial resistance: bacteria on the defence. Resistance stems from misguided efforts to try to sterilise our environment. PMID- 9727985 TI - Surveillance of antimicrobial resistance. Centralised surveys to validate routine data offer a practical approach. PMID- 9727986 TI - Lessons from New York's tuberculosis epidemic. Tuberculosis is a political as much as a medical problem-and so are the solutions. PMID- 9727987 TI - Which contacts of patients with meningococcal disease carry the pathogenic strain of Neisseria meningitidis? A population based study. AB - OBJECTIVES: To determine the prevalence of the pathogenic strain of Neisseria meningitidis in contacts of patients with meningococcal disease, and to determine which contact groups are likely to be carriers and warrant chemoprophylaxis. DESIGN: Population based study. SETTING: Norwegian county of Telemark. SUBJECTS: 1535 primary contacts of 48 patients with meningococcal disease, and 78 secondary contacts. INTERVENTIONS: Carriers of the pathogenic strain were treated with rifampicin. All household members and kissing contacts under 15 years of age were treated with oral penicillin. Contacts were taught to recognise the symptoms of meningococcal disease. RESULTS: In 27 of 48 cases investigated, contacts carrying the pathogenic strain of N meningitidis were found. A total of 42 such contacts were identified. Contacts were stratified into three classes according to the assumed closeness of contact with patients. In class 1 (household members and kissing contacts) the prevalence of the pathogenic strain was 12.4% (95% confidence interval 5.5% to 19.3%). In classes 2 and 3 the prevalence was 1.9% (0.9% to 3.4%) and 1.6% (0.14% to 3.1%). CONCLUSIONS: There is a high rate of carriage of the pathogenic strain of N meningitidis in patients' household members and kissing contacts, and this supports the practice of giving chemoprophylaxis to these contacts. The prevalence of carriage among other contacts is 2-3 times that found in the general population (0.7%); the benefits of chemoprophylaxis to these contacts may be marginal. PMID- 9727989 TI - Antibiotic resistance among enterococci causing endocarditis in the UK: analysis of isolates referred to a reference laboratory. PMID- 9727988 TI - Effect of preventive treatment for tuberculosis in adults infected with HIV: systematic review of randomised placebo controlled trials. AB - OBJECTIVE: To determine whether preventive treatment for tuberculosis in adults infected with HIV reduces the frequency of tuberculosis and overall mortality. DESIGN: Systematic review and data synthesis of randomised placebo controlled trials. MAIN OUTCOME MEASURES: Active tuberculosis, mortality, and adverse drug reaction requiring cessation of the study regimen. Outcomes stratified by status of purified protein derivative skin test. RESULTS: Four trials comprising 4055 adults from Haiti, Kenya, the United States, and Uganda were included. All compared isoniazid (6-12 months) with placebo, and one trial also compared multidrug treatment for 3 months with placebo. Mean follow up was 15-33 months. Overall, frequency of tuberculosis (relative risk 0.57, 95% confidence interval 0.41 to 0.79) was reduced in those receiving preventive treatment compared with placebo: mortality was not significantly reduced (0.93, 0.83 to 1.05). In subjects positive for purified protein derivative receiving preventive treatment, the risk of tuberculosis was reduced substantially (0.32, 0.19 to 0.51) and the risk of death was reduced moderately (0.73, 0.57 to 0.95) compared with those taking placebo. In adults negative for purified protein derivative receiving preventive treatment, the risk of tuberculosis (0.82, 0.50 to 1.36) and the risk of death (1.02, 0.89 to 1.17) were not reduced significantly. Adverse drug reactions were more frequent, but not significantly so, in patients receiving drug compared with placebo (1.45, 0.98 to 2.14). CONCLUSIONS: Preventive treatment given for 3-12 months protects against tuberculosis in adults infected with HIV, at least in the short to medium term. Protection is greatest in subjects positive for purified protein derivative, in whom death is also less frequent. Long term benefits remain to be shown. PMID- 9727990 TI - Multidrug resistant tuberculosis in France 1992-4: two case-control studies. PMID- 9727991 TI - Are amoxycillin and folate inhibitors as effective as other antibiotics for acute sinusitis? A meta-analysis. AB - OBJECTIVES: To examine whether antibiotics are indicated in treating uncomplicated acute sinusitis and, if so, whether newer and more expensive antibiotics with broad spectra of antimicrobial activity are more effective than amoxycillin or folate inhibitors. DESIGN: Meta-analysis of randomised trials. SETTING: Outpatient clinics. SUBJECTS: 2717 patients with acute sinusitis or acute exacerbation of chronic sinusitis from 27 trials. INTERVENTIONS: Any antibiotic versus placebo; amoxycillin or folate inhibitors versus newer, more expensive antibiotics. MAIN OUTCOME MEASUREMENTS: Clinical failures and cures. RESULTS: Compared with placebo, antibiotics decreased the incidence of clinical failures by half (risk ratio 0.54 (95% confidence interval 0.37 to 0.79)). Risk of clinical failure among 1553 randomised patients was not meaningfully decreased with more expensive antibiotics as compared with amoxycillin (risk ratio 0.86 (0.62 to 1.19); risk difference 0.9 fewer failures per 100 patients (1.4 more failures to 3.1 fewer failures per 100 patients)). The results were similar for other antibiotics versus folate inhibitors (risk ratio 1.01 (0.52 to 1.97)), but data were sparse (n=410) and of low quality. CONCLUSIONS: Amoxycillin and folate inhibitors are essentially as effective as more expensive antibiotics for the initial treatment of uncomplicated acute sinusitis. Small differences in efficacy may exist, but are unlikely to be clinically important. PMID- 9727992 TI - Understanding the culture of prescribing: qualitative study of general practitioners' and patients' perceptions of antibiotics for sore throats. AB - OBJECTIVES: To better understand reasons for antibiotics being prescribed for sore throats despite well known evidence that they are generally of little help. DESIGN: Qualitative study with semi-structured interviews. SETTING: General practices in South Wales. SUBJECTS: 21 general practitioners and 17 of their patients who had recently consulted for a sore throat or upper respiratory tract infection. MAIN OUTCOME MEASURES: Subjects' experience of management of the illness, patients' expectations, beliefs about antibiotic treatment for sore throats, and ideas for reducing prescribing. RESULTS: Doctors knew of the evidence for marginal effectiveness yet often prescribed for good relationships with patients. Possible patient benefit outweighed theoretical community risk from resistant bacteria. Most doctors found prescribing "against the evidence" uncomfortable and realised this probably increased workload. Explanations of the distinction between virus and bacterium often led to perceived confusion. Clinicians were divided on the value of leaflets and national campaigns, but several favoured patient empowerment for self care by other members of the primary care team. Patient expectations were seldom made explicit, and many were not met. A third of patients had a clear expectation for antibiotics, and mothers were more likely to accept non-antibiotic treatment for their children than for themselves. Satisfaction was not necessarily related to receiving antibiotics, with many seeking reassurance, further information, and pain relief. CONCLUSIONS: This prescribing decision is greatly influenced by considerations of the doctor patient relationship. Consulting strategies that make patient expectations explicit without damaging relationships might reduce unwanted antibiotics. Repeating evidence for lack of effectiveness is unlikely to change doctors' prescribing, but information about risk to individual patients might. Emphasising positive aspects of non-antibiotic treatment and lack of efficacy in general might be helpful. PMID- 9727993 TI - The development of new antimicrobial agents. PMID- 9727994 TI - What can be done about resistance to antibiotics? PMID- 9727995 TI - Antimicrobial resistance in developing countries. PMID- 9727996 TI - Surveillance of antimicrobial resistance--an international perspective. PMID- 9727997 TI - The epidemiology of antimicrobial resistance in hospital acquired infections: problems and possible solutions. PMID- 9727998 TI - Community acquired infections and bacterial resistance. PMID- 9727999 TI - The origins and molecular basis of antibiotic resistance. PMID- 9728000 TI - Antiviral drug resistance. PMID- 9728001 TI - Regulating the use of antibiotics in the community. PMID- 9728002 TI - Use of antimicrobial drugs in veterinary practice. PMID- 9728004 TI - Community based approaches to the control of multidrug resistant tuberculosis: introducing "DOTS-plus". PMID- 9728003 TI - Strategies for promoting judicious use of antibiotics by doctors and patients. PMID- 9728005 TI - Hypoxic responses in infants. Subjecting infants to low oxygen concentrations seems unethical. PMID- 9728006 TI - Ecological studies are a poor means of testing aetiological hypotheses. PMID- 9728007 TI - Studies must establish whether prolonged QTc interval in newly diagnosed type 1 diabetes is reversible. PMID- 9728008 TI - Helicobacter pylori and surgery. Bile reflux is important in eradicating Helicobacter pylori. PMID- 9728009 TI - Relation between birth weight and blood pressure is independent of maternal blood pressure. PMID- 9728010 TI - Screening for Chlamydia trachomatis. New methods are needed to assess the burden of illness from chlamydia. PMID- 9728011 TI - Other amputees are the greatest help in dealing with limb loss. PMID- 9728012 TI - Long term follow up of children with recurrent abdominal pain. Definition of recurrent abdominal pain was not applied. PMID- 9728013 TI - Central venous catheters and infection. Surveillance is effective in reducing catheter related sepsis. PMID- 9728014 TI - Effects of ambulance response times are being evaluated. PMID- 9728015 TI - Arnold appleby PMID- 9728016 TI - Some experience necessary PMID- 9728017 TI - The government's initiative may be stillborn PMID- 9728018 TI - Grapes PMID- 9728019 TI - Emerging infections PMID- 9728020 TI - Effective commissioning: lessons from purchasing in american managed care PMID- 9728021 TI - Bugs and drugs PMID- 9728022 TI - Chemoprophylaxis may be overused in meningococcal disease PMID- 9728023 TI - Preventive treatment for tuberculosis in adults infected with HIV does confer benefits PMID- 9728024 TI - Amoxycillin and folate inhibitors are as effective as newer antibiotics in acute sinusitis PMID- 9728025 TI - More evidence of ineffectiveness will not reduce antibiotic prescribing for sore throat PMID- 9728026 TI - Quarter of the enterococci responsible for endocarditis are resistant PMID- 9728027 TI - Multidrug resistant TB in france is associated with HIV PMID- 9728028 TI - Operating characteristics of six response distortion indicators for the personality assessment inventory. AB - The characteristics of six different indicators of response distortion on the Personality Assessment Inventory (PAI; Morey, 1991) were evaluated by having college students complete the PAI under positive impression management, malingering, and honest responding conditions. The six indicators were the PAI Positive Impression (PIM) and Negative Impression (NIM) scales, the Malingering and Defensiveness Indexes, and two discriminant functions, one developed by Cashel and the other by Rogers. Protocols of students asked to malinger were compared with those of actual clinical patients, while protocols of students asked to manage their impression in a positive direction were compared with those of students asked to respond honestly. Comparisons between groups were accomplished through the examination of effect sizes and receiver operating characteristic (ROC) curves. All six indicators demonstrated the ability to distinguish between actual and feigned responding. The Rogers function was particularly effective in identifying malingering. The Cashel function was less effective than other measures in identifying positive impression management, although it appears to also have promise as an indicator of malingering. PMID- 9728029 TI - The construct validity of the Tower of LondonDX as a measure of the executive functioning of ADHD children. AB - Currently, there are a limited number of measures that have been developed to assess childhood executive planning and problem-solving abilities. The present study represents ongoing efforts to determine the psychometric properties of the newly developed Tower of London Drexel (TOLDX), a measure of executive functions. Specifically, the construct-related validity of the TOLDX was investigated with a sample of attention-deficit hyperactivity disorder (ADHD) children (N = 129), 7 to 15 years of age. The performance of the children on a neuropsychological battery of measures sensitive to executive abilities, psychometric intelligence, and memory was subjected to maximum-likelihood factor analysis. A four-factor solution was extracted that best fit the selected variables. The TOLDX was found to produce the highest loading on an Executive Planning/Inhibition factor comprised of other executive measures. The Executive Planning/Inhibition measure was separate from factors of Executive Concept Formation/Flexibility, Psychometric Intelligence, and Memory. Moreover, the two executive factors were found to be correlated (r = .42) suggesting that each assesses, to varying degrees, related executive dimensions. The neuropsychological construct structure of the TOLDX is discussed in relationship with the temporal organization of behavior and frontal lobe functioning. PMID- 9728030 TI - Judgment of line orientation in patients with unilateral cerebrovascular lesions. AB - This study examined the relationship between visual neglect and ability to judge the angular orientation of lines in patients with unilateral cerebrovascualar lesions. Participants were 75 patients with right-hemisphere cerebrovascular lesions (RCVA) and 39 patients with left-hemisphere cerebrovascular lesions (LCVA). All were administered the Judgment of Line Orientation test (JLO). Results revealed that twice as many RCVA patients (56%) exhibited impairment on the JLO as did LCVA patients (26%). In RCVA patients, much lower performance was seen in patients with left visual neglect. Among LCVA patients, no relationship was found between JLO performance and the presence of visual neglect. Correlations between JLO scores and WAIS-R variables suggested that JLO performance may be related in part to general cognitive factors, as well as to other perceptual and spatial abilities. PMID- 9728031 TI - Novaco Anger Scale: reliability and validity within an adult criminal sample. AB - This study investigated the reliability and validity of the Novaco Anger Scale (NAS; Novaco, 1994) with two groups of correctional offenders, General Admissions and Violent Admissions. Predominantly White male offenders (N = 204), ranging in age from 18 to 69 years, participated in the study. One-month test-retest reliability for the General Admissions group ranged from .78 to .91 using both similar (paper-pencil) and dissimilar (computerized) retesting methods, with lower scores occurring on retest. Significantly lower scores were found for the Violent Admissions group as compared with the General Admissions group. Concurrent validity was examined in the Violent Admissions group using three anger/aggression measures and clinical ratings of eight anger dimensions. Stronger correlations with other similar anger measures than with negative affect indices revealed concurrent and discriminant validity. Implications for clinical use in an offender population are discussed. PMID- 9728032 TI - The use of the MMPI-2 for the assessment of depressive and psychotic disorders. AB - Considering the normative changes incorporated into the MMPI-2 and the introduction of content scales, this study examined its usefulness for the diagnosis and assessment of depression and psychosis in a heterogeneous sample of 264 psychiatric inpatients. We examined the mean group profiles and diagnostic efficiency of single scales at specified cutoff scores for these conditions. We also conducted cross-validated stepwise regression using all the basic and content scales as well as hierarchical regression examining the incremental validity of the basic and content scales. In general, the MMPI-2 profiles were found to be sensitive to group differences and the derived regression equations proved to be stable and fairly good at classification; but single scales were less useful for diagnosis. Changes in norms made the MMPI-2 more specific than the MMPI, and the introduction of new content scales offered considerable additional clinical information and incremental validity. PMID- 9728033 TI - Performance of children with traumatic brain injury on the Cognitive Assessment System. AB - Children and adolescents with traumatic brain injury (TBI) were compared to a matched sample of neurologically normal children and adolescents on several measures of cognitive processing. Each of the children in the TBI group had experienced a closed head injury of moderate to severe magnitude. Participants in both the TBI (n = 22) and control (n = 22) groups ranged in age from 9 to 17 years and lived in the midwestern United States. They were all administered the Cognitive Assessment System (Naglieri & Das, 1997a). Children with TBI earned significantly lower scores in the domains of Planning and Attention than the matched control group. Within-group comparisons showed that the TBI group s Planning and Attention scores were significantly lower than their Simultaneous and Successive scores. The results are consistent with previous literature demonstrating poor performance on measures of attention and executive functions among children who have experienced TBI. PMID- 9728034 TI - DSM-IV Cluster A personality disorder diagnoses among young adults with a 2-7-8 MMPI profile. AB - The present study evaluated the presence of DSM-IV personality disorders among young adults from a nonclinical setting who produced an MMPI 2-7-8 profile in comparison to a group of MMPI-defined controls. Categorical and dimensional analyses of personality disorders were evaluated. Participants in the 2-7-8 group (n = 20) received significantly more personality disorder diagnoses than did controls (n = 29), and 85% of these individuals received at least one Cluster A (Paranoid, Schizoid, Schizotypal) diagnosis in contrast to only 6.9% of controls (categorical analysis). The 2-7-8 group also received significantly more Cluster A diagnoses than Cluster B or C diagnoses. When dimensional analyses were applied (subclinical diagnoses), 95% of the 2-7-8 group evidenced Cluster A features. Comorbidity patterns were also evaluated; the most frequent comorbid diagnosis for the 2-7-8 group was Avoidant Personality Disorder (n = 8), consistent with Meehl s (1962, 1989, 1990) conceptualization of schizotypy. These results support the use of the MMPI 2-7-8 profile as an indicator of schizophrenia-related pathology within nonclinical samples of young adults. PMID- 9728035 TI - Evaluating a measure of the five-factor model of personality. AB - An evaluation is made of Goldberg s (1992) 100 Unipolar Markers of the five factor model of personality. The factor structure of these items in samples of older men from the Normative Aging Study and undergraduate students are examined, and both item transformation and consistency testing approaches are used to evaluate replications of the five-factor structure. Results show that the five factor structure is difficult to replicate in the sample of older men. While item transformations and sample trimming based on a consistency test did improve the quality of the replication in this older, nonstudent sample, both methods have serious drawbacks. The five-factor solution appeared in the student sample without sample trimming or data transformation. Additionally, in both student and nonstudent samples, oblique rotation resulted in inter-factor correlations relevant to more general issues in the study of trait structure. We conclude that the 100 Unipolar Markers may be unsuitable for use in older populations or with nonstudent samples. PMID- 9728036 TI - Epoxyeicosatrienoic acids relax airway smooth muscles and directly activate reconstituted KCa channels. AB - Epoxyeicosatrienoic acids (EETs) relax various smooth muscles by increasing outward K+ movement, but the molecular mode of action of EET regioisomers remains to be clarified. The effects of EETs were investigated on bovine airway smooth muscle tone and on reconstituted Ca2+-activated K+ (KCa) channels. 5,6-EET and 11, 12-EET induced dose-dependent relaxations of precontracted bronchial spirals. These effects were partly abolished by 10 nM iberiotoxin. Bilayer experiments have shown that 0.1-10 microM 11,12-EET produced up to fourfold increases in the open probability of KCa channels from the cis (extracellular) side by enhancing the mean open time constant and reducing the long closed time constant, without affecting the unitary conductance. EET-induced activations were blocked by 10 nM iberiotoxin. Addition of vehicles or other lipids as well as of GTP and guanosine 5'-O-(3-thiotriphosphate) in the absence of EET had no effect on channel activity. Thus EETs directly activate KCa channels from airway smooth muscle through an interaction with the extracellular face of the channel. We propose that EETs could represent candidate molecules as epithelium-derived hyperpolarizing factors. PMID- 9728037 TI - Direct modulation of tracheal Cl--channel activity by 5,6- and 11,12-EET. AB - Using microelectrode potential measurements, we tested the involvement of Cl- conductances in the hyperpolarization induced by 5,6- and 11,12 epoxyeicosatrienoic acid (EET) in airway smooth muscle (ASM) cells. 5,6-EET and 11,12-EET (0.75 microM) caused -5.4 +/- 1.1- and -3.34 +/- 0.95-mV hyperpolarizations, respectively, of rabbit tracheal cells (from a resting membrane potential of -53.25 +/- 0.44 mV), with significant residual repolarizations remaining after the Ca2+-activated K+ channels had been blocked by 10 nM iberiotoxin. In bilayer reconstitution experiments, we demonstrated that the EETs directly inhibit a Ca2+-insensitive Cl- channel from bovine ASM; 1 microM 5,6-EET and 1.5 microM 11,12-EET lowered the unitary current amplitude by 40 (n = 6 experiments) and 44.7% (n = 4 experiments), respectively. Concentration dependent decreases in channel open probability were observed, with estimated IC50 values of 0.26 microM for 5,6- and 1.15 microM for 11,12-EET. Furthermore, pharmacomechanical tension measurements showed that both regioisomers induced significant bronchorelaxations in epithelium-denuded ASM strips. These results suggest that 5,6- and 11,12-EET can act in ASM as epithelium-derived hyperpolarizing factors. PMID- 9728038 TI - Effect of hypoxic exposure on Na+/H+ antiport activity, isoform expression, and localization in endothelial cells. AB - Little is known about the effects of prolonged hypoxic exposure on membrane ion transport activity. The Na+/H+ antiport is an ion transport site that regulates intracellular pH in mammalian cells. We determined the effect of prolonged hypoxic exposure on human pulmonary arterial endothelial cell antiport activity, gene expression, and localization. Monolayers were incubated under hypoxic or normoxic conditions for 72 h. Antiport activity was determined as the rate of recovery from intracellular acidosis. Antiport isoform identification and gene expression were determined with RT-PCR and Northern and Western blots. Antiport localization and F-actin cytoskeleton organization were defined with immunofluorescent staining. Prolonged hypoxic exposure decreased antiport activity, with no change in cell viability compared with normoxic control cells. One antiport isoform [Na+/H+ exchanger isoform (NHE) 1] that was localized to the basolateral cell surface was present in human pulmonary arterial endothelial cells. Hypoxic exposure had no effect on NHE1 mRNA transcript expression, but NHE1 protein expression was upregulated. Immunofluorescent staining demonstrated a significant alteration of the F-actin cytoskeleton after hypoxic exposure but no change in NHE1 localization. These results demonstrate that the decrease in NHE1 activity after prolonged hypoxic exposure is not related to altered gene expression. The change in NHE1 activity may have important consequences for vascular function. PMID- 9728040 TI - Hypoxia-specific upregulation of calpain activity and gene expression in pulmonary artery endothelial cells. AB - The effects of exposure to hypoxia on the catalytic activity and mRNA expression of calpain, a calcium-regulated neutral cysteine protease, were examined in porcine pulmonary artery endothelial cells (PAECs). Specificity of the response to hypoxia was determined by comparing the effects of hypoxic exposure with exposure to oxidants such as nitrogen dioxide (NO2) and nitric oxide (NO), as well as to the sulfhydryl reactive chemical acrolein. Exposure of cells to hypoxia (0% O2) for 1 and 12 h significantly increased catalytic activity (P < 0.01 for both 1 and 12 h vs. control cells), as well as mRNA expression (P < 0.01 for 1 h and P < 0.05 for 12 h vs. control cells) of calpain. With more prolonged exposure to 24 h of hypoxia, calpain activity remained significantly elevated, whereas calpain mRNA expression returned to the control level. Calpain activities in cells exposed to NO2 [5 parts/million (ppm)] or NO (7.5 ppm) for 1 h or to acrolein (5 microM) for 1 and 24 h were unchanged. However, calpain activities in cells exposed to NO2 or NO for 24 h were significantly (P < 0.05) reduced compared with control cells. The hypoxia-induced increases in calpain mRNA content were prevented by the transcriptional inhibitor actinomycin D and by calpain inhibitor I. In addition, hypoxia increased the degradation of nuclear factor-kappaB (NF-kappaB) inhibitor IkappaB and enhanced the translocation of the p50 subunit of NF-kappaB to the nuclear membrane. Pretreatment with the calpain specific inhibitor E-64d prevented hypoxia-induced mRNA expression and degradation of IkappaBalpha, as well as translocation of p50 subunit to the nuclear membrane. These results demonstrate for the first time that hypoxia upregulates calpain activity and mRNA expression in PAECs and that the upregulation is specific to hypoxia. Upregulation appears to involve activation of the transcription factor NF-kappaB. PMID- 9728039 TI - Liposome-mediated transfection of fetal lung epithelial cells: DNA degradation and enhanced superoxide toxicity. AB - Cationic liposomes, 1:1 (mol/mol) 1,2-dioleoyldimethylammonium chloride-1,2 dioleoyl-sn-glycero-3-phosphoethanolamine, were used to transfect primary cultures of distal rat fetal lung epithelial cells with pCMV4-based plasmids. A DNA-to-lipid ratio of 1:10 to 1:15 (wt/wt) optimized DNA uptake over a 24-h exposure. At a fixed DNA-to-lipid ratio of 1:15, chloramphenicol acetyltransferase (CAT) reporter gene expression declined at lipid concentrations > 2.5 nmol/cm2 cell surface area, whereas DNA uptake remained concentration dependent. CAT expression peaked 48 h after removal of the liposome-DNA complex, declining thereafter. Reporter gene expression was increased, and supercoiled cDNA degradation was reduced by the addition of 0.2 mM nicotinamide and 10 microM chloroquine. Rat fetal lung epithelial cells transfected with two different expression cassettes had an increased susceptibility to superoxide-mediated cytotoxicity. This could be attributed to a nonspecific delivery of exogenous DNA or some other copurified factor. The DNA-dependent increase in superoxide mediated cytotoxicity, but not basal levels of cytotoxicity, was inhibited by the addition of 0.2 mM nicotinamide and 10 microM chloroquine. PMID- 9728041 TI - Interleukin-4 inhibits mitogen-induced proliferation of human airway smooth muscle cells in culture. AB - The increase in the amount of airway smooth muscle in the bronchial wall associated with asthma is partly due to hyperplasia. It is therefore important to determine which factors regulate growth and especially proliferation. In this study, we describe the effect of interleukin-4 (IL-4), a mast cell- and T lymphocyte-derived cytokine, on human airway smooth muscle proliferation as determined by [3H]thymidine uptake in the presence of fetal bovine serum (FBS), platelet-derived growth factor, basic fibroblast growth factor, and thrombin. IL 4 (5, 15, 50, and 150 ng/ml) significantly decreased 10% FBS-induced proliferation by 50, 73, 43, and 46%, respectively. The proliferative responses to platelet-derived growth factor (20 and 40 ng/ml), basic fibroblast growth factor (30 ng/ml), and thrombin (1 and 10 U/ml) were significantly reduced by 19, 21, 37, 36, and 57% respectively in the presence of 50 ng/ml of IL-4. We investigated the effect of IL-4 and other known inhibitors of smooth muscle proliferation, namely PGE2, heparin, and forskolin, on intracellular cAMP concentrations. IL-4 (50 ng/ml) and heparin (100 U/ml) did not alter intracellular cAMP levels when cells were treated with 1 or 10% FBS. PGE2 (1 microM) and forskolin (10 microM) significantly increased cAMP concentration above the control value in nonproliferating cells (1% FBS treated) by 7- and 37 fold, respectively. The effect of IL-4 (50 ng/ml), PGE2 (1 microM), and forskolin (10 microM) on cyclin D1 protein expression in 10% FBS-stimulated human airway smooth muscle cells was also examined. PGE2 and forskolin did not significantly inhibit cyclin D1 expression. However, IL-4 decreased cyclin D1 expression by 21%. These results provide evidence that IL-4 decreases human airway smooth muscle cell proliferation via a mechanism that is cAMP independent and mediated, in part, by a decrease in cyclin D1 protein expression. PMID- 9728042 TI - Upregulation of alveolar epithelial fluid transport after subacute lung injury in rats from bleomycin. AB - Alveolar epithelial fluid transport was studied 10 days after subacute lung injury had been induced with intratracheal bleomycin (0.75 U). An isosmolar Ringer lactate solution with 5% bovine serum albumin and 125I-labeled albumin as the alveolar protein tracer was instilled into the right lung; the rats were then studied for either 1 or 4 h. Alveolar fluid clearance was increased in bleomycin injured rats by 110% over 1 h and by 75% over 4 h compared with control rats (P < 0.05). The increase in alveolar fluid clearance was partially inhibited by amiloride (10(-3) M). Alveolar fluid clearance decreased toward normal levels in rats that were studied 60 days after bleomycin instillation. Remarkably, the measured increase in net alveolar fluid clearance occurred in the presence of a significant increase in alveolar epithelial permeability to protein. Moreover, the increase in alveolar epithelial fluid clearance occurred even though the mRNA for the alpha-subunit of the epithelial sodium channel was decreased in alveolar epithelial type II cells isolated from these rats. In addition, 22Na uptake by isolated alveolar epithelial type II cells from rats treated with bleomycin demonstrated a 52% decrease in uptake compared with type II cells from control rats. Morphological results demonstrated a significant hyperplasia of alveolar type II epithelial cells 10 days after bleomycin injury. Thus, these results provide evidence that proliferation of alveolar epithelial type II cells after acute lung injury may upregulate the transport capacity of the alveolar epithelium, even though the expression of epithelial sodium channels is reduced and the uptake of 22Na per cell is also reduced. These results may have clinical relevance for the resolution of alveolar edema in the subacute phase of lung injury. PMID- 9728043 TI - Prostanoids mediate IL-1beta-induced beta-adrenergic hyporesponsiveness in human airway smooth muscle cells. AB - We have previously reported that pretreatment of cultured human airway smooth muscle (HASM) cells with interleukin-1beta (IL-1beta) results in decreased beta adrenergic responsiveness. The purpose of this study was to determine whether prostanoids released as a result of cyclooxygenase-2 (COX-2) induction by IL 1beta contribute to this effect of the cytokine. Confluent serum-deprived HASM cells were studied in passages 4-7. IL-1beta (20 ng/ml for 22 h) reduced the ability of the beta-agonist isoproterenol (Iso) to decrease stiffness of HASM cells as measured by magnetic twisting cytometry. The effect of IL-1beta on Iso induced changes in cell stiffness was abolished by nonselective [indomethacin (Indo), 10(-6) M] and selective (NS-398, 10(-5) M) COX-2 inhibitors. Indo and NS 398 also inhibited both the increased basal cAMP and the decreases in Iso stimulated cAMP production induced by IL-1beta. IL-1beta (20 ng/ml for 22 h) caused an increase in both basal (15-fold) and arachidonic acid (AA)-stimulated (10-fold) PGE2 release. Indo blocked basal and AA-stimulated PGE2 release in both control and IL-1beta-treated cells. NS-398 also markedly reduced basal and AA stimulated PGE2 release in IL-1beta-treated cells but had no significant effect on AA-stimulated PGE2 release in control cells. Western blot analysis confirmed the induction of COX-2 by IL-1beta. Exogenously administered PGE2 (10(-7) M, 22 h) caused a significant reduction in the ability of Iso to decrease cell stiffness, mimicking the effects of IL-1beta. Cycloheximide (10 microg/ml for 24 h), an inhibitor of protein synthesis, also abolished the effects of IL-1beta on Iso-induced cell stiffness changes and cAMP formation. In summary, our results indicate that IL-1beta significantly increases prostanoid release by HASM cells as a result of increased COX-2 expression. The prostanoids appear to contribute to beta-adrenergic hyporesponsiveness, perhaps by heterologous desensitization of the beta2 receptor. PMID- 9728044 TI - Cigarette smoke increases amosite asbestos fiber binding to the surface of tracheal epithelial cells. AB - Binding of asbestos fibers to the cell surface appears to be important in the initiation of intracellular signaling events as well as in initiation of particle uptake by the cell. We have previously shown that cigarette smoke increases the uptake of asbestos fibers by tracheal epithelial cells in explant culture. Whether smoke acts by increasing surface binding of fibers is not known. In this study, we exposed rat tracheal explants to air or cigarette smoke and then to a suspension of amosite asbestos. Explants were harvested after 1 or 24 h of dust exposure and washed by repeated dips in culture medium to remove loosely bound fibers, and the number of fibers adhering to the apical cell surfaces was determined by scanning electron microscopy. Smoke-exposed explants retained significantly more surface fibers than air-exposed explants. After four washes, binding levels were similar at 1 and 24 h. The smoke effect was still present when incubations were carried out at 4 degrees C, but binding was decreased approximately 25%. Preincubation of the asbestos fibers with iron chloride to increase surface iron increased fiber binding in both air- and smoke-exposed explants, whereas preincubation of the fibers with the iron chelator deferoxamine decreased binding after air exposure and completely eliminated the smoke effect. Inclusion of mannitol or catalase in the medium or preincubation of the explants with GSH produced decreases in binding of 10-25% in air-exposed explants and generally greater decreases in smoke-exposed explants. We conclude that 1) amosite binding is a very rapid process that does not require active cellular metabolism, 2) cigarette smoke increases adhesion of fibers to the epithelial surfaces, and 3) iron on the asbestos fiber appears to play an important role in binding, probably through an active oxygen species-mediated process. PMID- 9728046 TI - Ionic mechanisms underlying electrical slow waves in canine airway smooth muscle. AB - In canine bronchial smooth muscle (BSM), spasmogens evoke oscillations in membrane potential ("slow waves"). The depolarizing phase of the slow waves is mediated by voltage-dependent Ca2+ channels; we examined the roles played by Cl- and K+ currents and Na+-K+-ATPase activity in mediating the repolarizing phase. Slow waves were evoked using tetraethylammonium (25 mM) in the presence or absence of niflumic acid (100 microM; Cl- channel blocker) or ouabain (10 microM; block Na+-K+-ATPase) or after elevating external K+ concentration ([K+]) to 36 mM (to block K+ currents); curve fitting was performed to quantitate the rates of rise/fall and frequency under these conditions. Slow waves were markedly slowed, and eventually abolished, by niflumic acid but were unaffected by ouabain or high [K+]. Electrically evoked slow waves were also blocked in similar fashion by niflumic acid. We conclude that the repolarization phase is mediated by Ca2+ dependent Cl- currents. This information, together with our earlier finding that the depolarizing phase is due to voltage-dependent Ca2+ current, suggests that slow waves in canine BSM involve alternating opening and closing of Ca2+ and Cl- channels. PMID- 9728045 TI - Nitric oxide inhalation transiently elevates pulmonary levels of cGMP, iNOS mRNA, and TNF-alpha. AB - The initial pulmonary vasodilation that occurs during nitric oxide (. NO) inhalation does not appear to be maintained chronically in many cases. . NO may acutely relax vascular smooth muscle by increasing levels of guanosine 3',5' cyclic monophosphate (cGMP), tumor necrosis factor (TNF)-alpha, and inducible nitric oxide synthase (iNOS) while decreasing levels of lipid peroxidation. It was hypothesized that the acute . NO-induced changes in cGMP, TNF-alpha, iNOS, and lipid peroxidation, all of which may mediate vasodilation, are transient rather than sustained. Lungs from rats kept in chambers containing 6 parts/million . NO for 1 h, 1 day, or 1 wk were analyzed for levels of . NO induced vasodilatory mediators. Pulmonary cGMP, iNOS mRNA, and TNF-alpha were increased 1 h after . NO exposure but decreased to control values at later times. Levels of malonyl dialdehyde, an indicator of lipid peroxidation, were decreased at all times during . NO inhalation. As a whole, the data suggest that in lungs the vasodilatory mediators cGMP, iNOS, and TNF-alpha are only acutely and transiently elevated during inhalation of . NO, consistent with the initially positive clinical response to inhaled . NO that deteriorates over time. PMID- 9728047 TI - IL-8 is one of the major chemokines produced by monkey airway epithelium after ozone-induced injury. AB - A rhesus monkey interleukin (IL)-8 cDNA clone with >94% homology to the human IL 8 gene was isolated by differential hybridization from a cDNA library of distal airways after ozone inhalation. In situ hybridization and immunohistochemistry showed increased IL-8 mRNA and protein levels in epithelial cells at 1 h but not at 24 h after inhalation of ozone. The appearance of IL-8 in airway epithelial cells correlated well with neutrophil influx into airway epithelia and lumens. Air-liquid interface cultures of tracheobronchial epithelial cells were exposed to ozone in vitro. We observed a transient increase in IL-8 secretion in culture medium immediately after ozone exposure and a dose-dependent increase in IL-8 secretion and mRNA production. In vitro neutrophil chemotaxis showed a parallel dose and time profile to epithelial cell secretion of IL-8. Treatment with anti IL-8 neutralizing antibody blocked >80% of the neutrophil chemotaxis in vitro. These results suggest that IL-8 is a key chemokine in acute ozone-induced airway inflammation in primates. PMID- 9728048 TI - E-selectin expression in human endothelial cells by TNF-alpha-induced oxidant generation and NF-kappaB activation. AB - Because reactive oxygen species (ROS) can function as second messengers and regulate the activation of the transcription factor nuclear factor (NF)-kappaB, we investigated the possible role of tumor necrosis factor-alpha (TNF-alpha) induced ROS generation in endothelial cells in signaling E-selectin gene transcription. We demonstrated that stimulation of human pulmonary artery endothelial cells with TNF-alpha (100 U/ml) resulted in ROS production using the oxidant-sensitive dye 5 (and 6)-carboxy-2',7'-dichlorodihydrofluorescein diacetate bis(acetoxymethyl)ester. Pretreatment with N-acetyl-L-cysteine (NAC) or pyrrolidine dithiocarbamate (PDTC) for 0.5 h inhibited TNF-alpha-induced generation of ROS as well as activation of NF-kappaB and E-selectin mRNA and the cell surface protein expression. These findings indicate that TNF-alpha induces NF-kappaB activation and the resultant E-selectin gene expression by a pathway that involves formation of ROS and that E-selectin expression can be inhibited by the antioxidant action of NAC or PDTC. The results support the hypothesis that generation of ROS in endothelial cells induced by proinflammatory cytokines such as TNF-alpha is a critical signal mediating E-selectin expression. PMID- 9728049 TI - Mesenchymal determination of mechanical strain-induced fetal lung cell proliferation. AB - Fetal breathing movements play an important role in normal fetal lung growth. We have previously shown that an intermittent mechanical strain regimen (60 cycles/min, 15 min/h), simulating normal fetal breathing movements, stimulated growth of mixed fetal rat lung cells in organotypic culture. In the present study, we examined the individual responses of the two major fetal lung cell types, fibroblasts and epithelial cells, to mechanical strain. Also, we investigated the effect of mesenchymal-epithelial interactions on strain-induced cell proliferation during fetal lung development. Fibroblasts and epithelial cells from day 18 to day 21 fetal rat lung (term = 22 days), cultured alone or as various recombinants, were subjected to either a 48-h static culture or to strain, and DNA synthesis was measured. Both cell types responded individually to strain with enhanced DNA synthesis throughout late fetal lung development. Independent of the recombination ratio, there was no additive response to strain when fibroblasts and epithelial cells from the same gestation were recombined. In contrast, strain-induced DNA synthesis was suppressed when cells from different gestations were recombined. The ontogenic response pattern of recombinants to mechanical strain was similar to that of fibroblasts but not of epithelial cells. Strain-induced proliferation increased and peaked at the early canalicular stage of lung development at 19 days of gestation and declined thereafter. We conclude that strain-enhanced growth of the fetal lung is gestation dependent and that the gestational response to mechanical force is regulated by the mesenchyme. PMID- 9728050 TI - Activation of MAPKs in human bronchial epithelial cells exposed to metals. AB - We have previously shown that in vitro exposure to metallic compounds enhances expression of interleukin (IL)-6, IL-8, and tumor necrosis factor-alpha in human bronchial epithelial cells. To characterize signaling pathways involved in metal induced expression of inflammatory mediators and to identify metals that activate them, we studied the effects of As, Cr, Cu, Fe, Ni, V, and Zn on the mitogen activated protein kinases (MAPK) extracellular receptor kinase (ERK), c-Jun NH2 terminal kinase (JNK), and P38 in BEAS cells. Noncytotoxic concentrations of As, V, and Zn induced a rapid phosphorylation of MAPK in BEAS cells. Activity assays confirmed marked activation of ERK, JNK, and P38 in BEAS cells exposed to As, V, and Zn. Cr and Cu exposure resulted in a relatively small activation of MAPK, whereas Fe and Ni did not activate MAPK under these conditions. Similarly, the transcription factors c-Jun and ATF-2, substrates of JNK and P38, respectively, were markedly phosphorylated in BEAS cells treated with As, Cr, Cu, V, and Zn. The same acute exposure to As, V, or Zn that activated MAPK was sufficient to induce a subsequent increase in IL-8 protein expression in BEAS cells. These data suggest that MAPK may mediate metal-induced expression of inflammatory proteins in human bronchial epithelial cells. PMID- 9728051 TI - Surfactant protein B processing in human fetal lung. AB - Surfactant protein B (SP-B8), an 8-kDa hydrophobic protein essential for surfactant and normal lung function, is produced from the intracellular processing of preproSP-B. To characterize SP-B processing in human type 2 cells, we used human fetal lung in explant culture and polyclonal antibodies to human SP B8 (Phe201-Met279) and to specific epitopes within the NH2- and COOH-terminal propeptide domains (Ser145-Leu160, Gln186-Gln200, and Gly284-Ser304). Western blot analysis revealed a novel intermediate at approximately 9 kDa, representing mature SP-B8, with a residual NH2-terminal peptide of approximately 10 amino acids. Pulse-chase studies showed a precursor-product relationship between the 9- and 8-kDa forms. During differentiation of type 2 cells in explant culture, the rate of proSP-B conversion to 25-kDa intermediate remained constant, whereas the rate of 25-kDa intermediate conversion to SP-B8 increased, resulting in a net increase in tissue SP-B8. Dexamethasone did not affect the rate of proSP-B processing but markedly enhanced the rate of SP-B8 accumulation. We conclude that NH2-terminal propeptide cleavage of proSP-B is a multistep process and that more distal processing events are rate limiting and both developmentally and hormonally regulated. PMID- 9728052 TI - Prolonged high intermittent positive-pressure ventilation induces airway remodeling and reactivity in young rats. AB - We postulated that prolonged exposure to intermittent positive-pressure ventilation (IPPV) with high pressure (HIPPV) alone without hyperoxia promotes the development of airway hyperresponsiveness and remodeling. To test this hypothesis, young rats were ventilated under halothane anesthesia with HIPPV (maximum inspiratory pressure at 32-35 cmH2O in 70% nitrous oxide and 30% O2) for 3.5-4 h daily for 6 days. Control rats were ventilated with low IPPV (maximum inspiratory pressure < 13 cmH2O) during the same time period with the same gas mixture. With the use of tracheal rings isolated from these rats and a setup in tissue baths, contractile responses to carbachol (10(-6) to 10(-2) mM), 5 hydroxytryptamine (5-HT; 10(-9) to 10(-5) mM) and KCl (1-100 mM) were examined isometrically. In tracheal rings from HIPPV rats compared with low-pressure IPPV rats, the concentration tension curves showed a significantly enhanced response to all agonists (P < 0.005). Sensitivity to carbachol, 5-HT, and KCl was also significantly increased (P < 0.05) compared with control rats as evidenced by decreases in EC50. Maximum tension (reactivity) to 5-HT and KCl in the HIPPV group increased significantly (P < 0.05), and there was a trend (P = 0.07) toward increased reactivity to carbachol in this group as well. Histological examinations of tracheal rings demonstrated epithelial squamous metaplasia in the HIPPV group. Morphometric studies demonstrated tracheal smooth muscle thickening (P < 0.05) without changes in the thickness of the mucosa or the lamina propria. When contractile responses were normalized for the smooth muscle cross-sectional area (i.e., stress), reactivity to all contractile agents was reduced, whereas reactivity to 5-HT still demonstrated significant increase (P < 0.005). Sensitivity of tracheal segments to all three agents was not affected by this normalization. These findings suggest that prolonged exposure to HIPPV without hyperoxia and the resultant overdistension of lung tissues (volutrauma) induced airway remodeling and airway hyperreactivity. PMID- 9728054 TI - Induction of ferritin and heme oxygenase-1 by endotoxin in the lung. AB - Heme oxygenase (HO)-1 expression is increased by forms of oxidative stress that also induce ferritin. Even though this could result from release of iron by heme degradation, we hypothesized that ferritin expression in the lung after endotoxin [lipopolysaccharide (LPS)] would occur independently of HO-1 because iron sequestration is an important response to infection. We tested this hypothesis by instilling saline or LPS (1 mg) into lungs of rats and measuring ferritin expression, HO-1 expression and activity, and HO-1 and ferritin mRNAs at different times. Lungs were also inflation fixed for immunohistochemistry for HO 1 and ferritin. Studies were performed with and without the HO inhibitor tin protoporphyrin. Ferritin and HO-1 labeling were minimal (macrophages only) in control lungs. By 4 h after LPS instillation, ferritin staining was present in bronchial epithelium and macrophages, became diffuse at 16 h, and was nearly gone by 48-72 h. HO-1 was detectable in macrophages 4 and 16 h after LPS instillation, increased in macrophages and bronchial epithelium at 24 h, and diffusely increased in bronchial epithelium and the alveolar region at 48-72 h. Lung ferritin content increased significantly by 4 h and peaked at 16 h before declining. HO-1 protein was present by Western blot in control lung, stable at 4 h, and increased by 24 h after LPS instillation, whereas HO enzyme activity had increased by 4 h after LPS instillation. After complete inhibition of HO enzyme activity with tin protoporphyrin, ferritin increased threefold at 4 h and sixfold at 24 h after LPS instillation. HO-1 mRNA increased by 4 h and was sustained at 24 h, whereas ferritin mRNA did not change after LPS instillation. These results indicate that intratracheal LPS rapidly induces ferritin protein in the lung independently of its mRNA synthesis or HO enzyme activity. LPS induces HO-1 mRNA, which is followed by increased expression of protein. PMID- 9728053 TI - Store-operated calcium entry promotes shape change in pulmonary endothelial cells expressing Trp1. AB - Activation of Ca2+ entry is known to produce endothelial cell shape change, leading to increased permeability, leukocyte migration, and initiation of angiogenesis in conduit-vessel endothelial cells. The mode of Ca2+ entry regulating cell shape is unknown. We hypothesized that activation of store operated Ca2+ channels (SOCs) is sufficient to promote cell shape change necessary for these processes. SOC activation in rat pulmonary arterial endothelial cells increased free cytosolic Ca2+ that was dependent on a membrane current having a net inward component of 5.45 +/- 0.90 pA/pF at -80 mV. Changes in endothelial cell shape accompanied SOC activation and were dependent on Ca2+ entry-induced reconfiguration of peripheral (cortical) filamentous actin (F actin). Because the identity of pulmonary endothelial SOCs is unknown, but mammalian homologues of the Drosophila melanogaster transient receptor potential (trp) gene have been proposed to form Ca2+ entry channels in nonexcitable cells, we performed RT-PCR using Trp oligonucleotide primers in both rat and human pulmonary arterial endothelial cells. Both cell types were found to express Trp1, but neither expressed Trp3 nor Trp6. Our study indicates that 1) Ca2+ entry in pulmonary endothelial cells through SOCs produces cell shape change that is dependent on site-specific rearrangement of the microfilamentous cytoskeleton and 2) Trp1 may be a component of pulmonary endothelial SOCs. PMID- 9728055 TI - Lung endothelial cell proliferation in normal and pulmonary hypertensive neonatal calves. AB - Tremendous changes in pressure and flow occur in the pulmonary and systemic circulations after birth, and these hemodynamic changes should markedly affect endothelial cell replication. However, in vivo endothelial replication rates in the neonatal period have not been reported. To label replicating endothelial cells, we administered the thymidine analog bromodeoxyuridine to calves approximately 1, 4, 7, 10, and 14 days old before they were killed. Because we expected the ratio of replicating to nonreplicating cells to vary with vascular segment, we examined the main pulmonary artery, a large elastic artery, three sizes of intrapulmonary arteries, the aorta, and the carotid artery. In normoxia for arteries < 1,500 micron, approximately 27% of the endothelial cells were labeled on day 1 but only approximately 2% on day 14. In the main pulmonary artery, only approximately 4% of the endothelial cells were labeled on day 1 and approximately 2% on day 14. In contrast, in the aorta, approximately 12% of the endothelial cells were labeled on day 1 and approximately 2% on day 14. In chronically hypoxic animals, only approximately 14% of the endothelial cells were labeled on day 1 in small lung arteries and approximately 8% were still labeled on day 14. We conclude that the postnatal circulatory adaptation to extrauterine life includes significant changes in endothelial cell proliferation that vary dramatically with time and vascular location and that these changes are altered in chronic hypoxia. PMID- 9728056 TI - Differential regulation of eotaxin expression by TNF-alpha and PMA in human monocytic U-937 cells. AB - Regulation of eotaxin expression was investigated in U-937 cells, a human monocyte-like cell line. Eotaxin mRNA was induced by tumor necrosis factor-alpha (TNF-alpha; 0.1-100 ng/ml) and phorbol 12-myristate 13-acetate (PMA; 0.01-1 microM). PMA-induced eotaxin mRNA expression was of greater magnitude and was maximal at a later time point than TNF-alpha-induced expression (16 h vs. 2 h after stimulation), which was consistent with eotaxin protein expression detected by immunocytochemistry. Dexamethasone (0.01-10 microM) decreased eotaxin mRNA expression in both TNF-alpha- and PMA-stimulated U-937 cells. PMA-induced eotaxin mRNA expression was inhibited by cycloheximide (10 microg/ml), whereas TNF-alpha induced expression was not. The protein kinase C (PKC) inhibitor staurosporine (10-50 nM) inhibited PMA-induced eotaxin mRNA expression, whereas TNF-alpha induced expression was enhanced by this reagent. These results suggest that eotaxin expression can be induced by more than one mechanism: the PMA-triggered pathway is mediated by PKC activation and requires new protein synthesis, whereas the TNF-alpha-triggered pathway is independent of PKC and protein synthesis. TNF alpha- and PMA-induced pathways are both associated with nuclear factor-kappaB, because its binding activity was enhanced in the presence of these stimuli, and both pathways were limited by its inhibitor, diethyldithiocarbamate. PMID- 9728057 TI - Interleukin-6 production in hemorrhagic shock is accompanied by neutrophil recruitment and lung injury. AB - Hemorrhagic shock (HS) initiates an inflammatory cascade that includes the production of cytokines and recruitment of neutrophils (PMN) and may progress to organ failure, inducing acute respiratory distress syndrome (ARDS). To examine the hypothesis that interleukin-6 (IL-6) contributes to PMN infiltration and lung damage in HS, we examined the lungs of rats subjected to unresuscitated and resuscitated HS for the production of IL-6 and activation of Stat3. Using semiquantitative RT-PCR, we found a striking increase in IL-6 mRNA levels only in resuscitated HS, with peak levels observed 1 h after initiation of resuscitation. Increased IL-6 protein expression was localized to bronchial and alveolar cells. Electrophoretic mobility shift assay of protein extracts from shock lungs exhibited an increase in Stat3 activation with kinetics similar to IL-6 mRNA. In situ DNA binding assay determined Stat3 activation predominantly within alveoli. Intratracheal instillation of IL-6 alone into normal rats resulted in PMN infiltration into lung interstitium and alveoli, marked elevation of bronchoalveolar lavage cellularity, and increased wet-to-dry ratio. These findings indicate that IL-6 production and Stat3 activation occur early in HS and may contribute to PMN-mediated lung injury, including ARDS after HS. PMID- 9728058 TI - Laminin 2 attachment selects myofibroblasts from fetal mouse lung. AB - Laminins (LNs) are extracellular matrix glycoproteins that are involved in cell adhesion, proliferation, and differentiation. So far, 11 LN variants (LN1 to LN11) have been described. In the lung, at least six LN variants have been identified. However, only the role of LN1 has been characterized to any extent. In this study, we hypothesized that the LN2 variant may play a role during lung development. We identified, by RT-PCR analysis, that the alpha2-chain mRNA of LN2 is expressed during mouse lung development. LN2 adhesion assays were then performed with cells from fetal mouse lung primary cultures. Our results showed that a specific subpopulation of fetal lung cells that expressed vimentin, alpha smooth muscle actin, and desmin attached onto LN2, whereas the cells that did not adhere to LN2 as well as the total cell population were able to adhere readily on fibronectin. Cell attachment onto LN2 was inhibited by EDTA. In addition, we demonstrated, by RT-PCR and Western analysis, that the LN2-adherent cells autoexpressed the alpha2-chain of LN2. In the late pseudoglandular period, LN2 was localized by immunohistochemistry in the basement membrane of airways and blood vessels and around mesenchymal cells. We conclude that LN2 is expressed during lung development and that a specific subpopulation of fetal lung mesenchymal cells expressing a myofibroblastic phenotype can be selected by attachment to LN2 in primary culture. These findings lead us to speculate that LN2 may play a key role in the cell biology of myofibroblasts during lung development. PMID- 9728059 TI - Involvement of lung interstitial proteoglycans in development of hydraulic- and elastase-induced edema. AB - We extracted and isolated proteoglycans from lung tissue samples obtained from three groups of anesthetized rabbits: 1) control animals (C; n = 8) killed by overdose after 180 min; 2) animals receiving an intravenous saline infusion (S; n = 4, 1.5 ml . kg-1 . min-1) for 180 min; 3) animals receiving an intravenous bolus of 200 microg of pancreatic elastase (E; n = 4), killed after 200 min. The lung dry weight-to-wet weight ratio in the three groups was 5.2 +/- 0.2, 6.0 +/- 0.4, and 5.6 +/- 0.5, respectively. Gel-filtration analysis showed a massive fragmentation of the versican family of the extracellular matrix (ECM) in the S groups and a marked degradation of heparan sulfate-containing proteoglycans, including perlecan of the basement membrane, in the E group. The binding properties of total proteoglycans to other ECM components were lowered in both groups relative to control. The decrease in proteoglycan binding was more pronounced for collagen type IV in the E group relative to C (-93.5%, P < 0.05) and for hyaluronic acid in the S groups (-85.8%, P < 0.05). These findings suggest that elastase treatment produces a major degree of damage to the organization of basement membrane, whereas saline loading affects more markedly the architecture of interstitial ECM. Qualitative zymography performed on lung extracts showed increased gelatinase activities in both S and E groups, providing direct evidence that the activation of tissue proteinases may play a role in acute lung injury. PMID- 9728060 TI - Effects of conjugated linoleic acid on body fat and energy metabolism in the mouse. AB - Conjugated linoleic acid (CLA) is a naturally occurring group of dienoic derivatives of linoleic acid found in the fat of beef and other ruminants. CLA is reported to have effects on both tumor development and body fat in animal models. To further characterize the metabolic effects of CLA, male AKR/J mice were fed a high-fat (45 kcal%) or low-fat (15 kcal%) diet with or without CLA (2.46 mg/kcal; 1.2 and 1.0% by weight in high- and low-fat diets, respectively) for 6 wk. CLA significantly reduced energy intake, growth rate, adipose depot weight, and carcass lipid and protein content independent of diet composition. Overall, the reduction of adipose depot weight ranged from 43 to 88%, with the retroperitoneal depot most sensitive to CLA. CLA significantly increased metabolic rate and decreased the nighttime respiratory quotient. These findings demonstrate that CLA reduces body fat by several mechanisms, including a reduced energy intake, increased metabolic rate, and a shift in the nocturnal fuel mix. PMID- 9728061 TI - Coordination of autonomic and behavioral thermoregulatory responses during exposure to a novel stimulus in rats. AB - The induction of psychological stress in rats is accompanied by an elevation of core temperature. Our experiments were carried out to determine whether the latency, duration, magnitude, or effector mechanisms of the core temperature response to psychological stress would be altered when rats were allowed to use behavioral as well as autonomic thermoregulation. Core temperature, oxygen consumption, and ambient temperature were measured in adult rats before and after handling and a sham intraperitoneal injection. Seven rats were studied in a thermocline (gradient of 7 to 42 degrees C) and eight rats were studied in a metabolic chamber (25 degrees C). The rats studied in the thermocline selected a warm ambient temperature following the sham intraperitoneal injection and exhibited an increase in core temperature of shorter latency, greater magnitude, and greater duration than those studied in the metabolic chamber. The rats studied in the metabolic chamber exhibited an oxygen consumption response of greater magnitude and duration than the animals studied in the thermocline. Thus the characteristics in addition to the effector mechanisms of the core temperature response to psychological stress are altered when rats are allowed to use behavioral as well as autonomic thermoregulatory effectors. PMID- 9728062 TI - Decompression sickness risk in rats by microbial removal of dissolved gas. AB - We present a method for reducing the risk of decompression sickness (DCS) in rats exposed to high pressures of H2. Suspensions of the human colonic microbe Methanobrevibacter smithii were introduced via a colonic cannula into the large intestines of the rats. While the rats breathed H2 in a hyperbaric chamber, the microbe metabolized some of the H2 diffusing into the intestine, converting H2 and CO2 to methane and water. Rate of release of methane from the rats, which was monitored by gas chromatography, varied with chamber H2 pressure. This rate was higher during decompression than during compression, suggesting that during decompression the microbe was metabolizing H2 stored in the rats' tissues. Rats treated with M. smithii had a 25% (5 of 20) incidence of DCS, which was significantly lower (P < 0.01) than the 56% (28 of 50) incidence of untreated controls, brought on by a standardized compression and decompression sequence. Thus using a microbe in the intestine to remove an estimated 5% of the body burden of H2 reduced DCS risk by more than one-half. This method of biochemical decompression may potentially facilitate human diving. PMID- 9728063 TI - Increased in vitro fatty acid supply and cellular transport capacities in cold acclimated ducklings (Cairina moschata). AB - In cold-acclimated (CA) birds, lipids play a crucial role in regulatory thermogenesis by acting both as substrates for and activators of thermogenic processes. The capacity to supply lipids to thermogenic tissues, which could limit cold thermogenesis, was assessed in CA ducklings (5 wk old, 4 degrees C) and compared with thermoneutral controls (TN, 25 degrees C). In CA ducklings, basal lipolytic activity of adipose tissue fragments was higher (202 +/- 9 vs. 130 +/- 14 nmol glycerol released . 100 mg tissue-1 . h-1, +55%) than in TN controls, while glucagon had a much higher stimulatory effect (+140 to +500% depending on dose). This was consistent with increased plasma levels of nonesterified fatty acids (FA, +57%) and glycerol (+31%) in vivo. In vitro endothelial lipase activity per organ was higher in CA than in TN ducklings in red gastrocnemius muscle (6.3 +/- 0.6 vs. 3.5 +/- 0.3 microeq nonesterified FA released per hour, +80%) and liver (+55%). The intracellular FA-binding capacity of (12-18 kDa) proteins was higher in gastrocnemius muscle (+43%) and liver (+74%) from CA ducklings. In gastrocnemius, it was linked to a higher content (21 +/- 2 vs. 15 +/- 2 microg/mg protein, +37%) of an intracellular 15.4-kDa FA binding protein. These in vitro results indicate that coordinated increases in FA supply from adipose tissue, cellular uptake of lipoprotein-derived FA, and intracellular FA transport capacity occur in CA ducklings endowed with higher thermogenic capacity and cold endurance. PMID- 9728064 TI - AVP mediates the attenuated febrile response to administration of PGE1 in rats near term of pregnancy. AB - Rats have an attenuated febrile response to intracerebroventricular injection of PGE1 near the term of pregnancy, the mechanism of which is unknown. The present experiments were carried out to test the hypothesis that arginine vasopressin (AVP), functioning as an endogenous antipyretic substance in the central nervous system, mediates this attenuated febrile response. The febrile response to intracerebroventricular injection of 0.2 microg PGE1 was determined in pregnant and nonpregnant rats after an intracerebroventricular injection of either vehicle or a vasopressin V1-receptor antagonist. After intracerebroventricular administration of vehicle, intracerebroventricular administration of 0.2 microg PGE1 produced significant increases in core temperature in both nonpregnant and pregnant animals. The increase in core temperature, however, was attenuated both in magnitude and duration in pregnant compared with nonpregnant animals. After intracerebroventricular administration of a vasopressin V1-receptor antagonist, intracerebroventricular administration of 0.2 microg PGE1 produced significant increases in core temperature that were similar in nonpregnant and pregnant animals. Our data support the hypothesis that a pregnancy-related activation of AVP as an endogenous antipyretic substance in the central nervous system attenuates the febrile response to intracerebroventricular administration of PGE1 near term of pregnancy in rats. PMID- 9728065 TI - Excretory transport of xenobiotics by dogfish shark rectal gland tubules. AB - Marine elasmobranch rectal gland is a specialized, osmoregulatory organ composed of numerous blind-ended, branched tubules emptying into a central duct. To date, NaCl excretion has been its only described function. Here we use isolated rectal gland tubule fragments from dogfish shark (Squalus acanthias), fluorescent xenobiotics, and confocal microscopy to describe a second function, xenobiotic excretion. Isolated rectal gland tubules rapidly transported the fluorescent organic anion sulforhodamine 101 from bath to lumen. Luminal accumulation was concentrative, saturable, and inhibited by cyclosporin A (CSA), chlorodinitrobenzene, leukotriene C4, and KCN. Inhibitors of renal organic anion transport (probenecid, p-aminohippurate), organic cation transport (tetraethylammonium and verapamil), and P-glycoprotein (verapamil) were without effect. Cellular accumulation of sulforhodamine 101 was not concentrative, saturable, or inhibitable. Rectal gland tubules did not secrete fluorescein, daunomycin, or a fluorescent CSA derivative. Finally, frozen rectal gland sections stained with an antibody to a hepatic canalicular multispecific organic anion transporter (cMOAT or MRP2) showed heavy and specific staining on the luminal membrane of the epithelial cells. We conclude that rectal gland is capable of active and specific excretion of xenobiotics and that such transport is mediated by a shark analog of MRP2, an ATP-driven xenobiotic transporter, but not by P-glycoprotein. PMID- 9728066 TI - The thromboxane A2 mimetic U-46619 inhibits somatomotor activity via a vagal reflex from the lung. AB - Vagal reflexes from the heart and lungs elicit autonomic as well as somatomotor responses. The purpose of the present investigation was to determine whether the inflammatory mediator thromboxane A2 inhibits the knee-jerk reflex via a vagally mediated reflex from either the heart or the lung. The thromboxane A2 mimetic U 46619 (0. 8 +/- 0.08 microg/kg) was injected through a catheter placed near the right atrium (n = 11), near the aortic arch (n = 7), or into the pericardial sac (n = 4) in 11 chloralose-anesthetized cats. The knee-jerk reflex, elicited by striking the patellar tendon with a solenoid-driven hammer, was used to evaluate somatomotor activity. The mean maximum tension produced by the knee-jerk reflex was 306 +/- 21 g (range 154-471 g). Intravenous U-46619 injection inhibited the knee-jerk reflex by 25 +/- 6% and increased peak systolic pressure 53 +/- 7 mmHg on average. Bilateral cervical vagotomy abolished the somatomotor inhibition but did not reduce the pressor response. Intra-arterial U-46619 injection inhibited the knee-jerk reflex in two of seven cats and increased peak systolic pressure by 41 +/- 11 mmHg. Vagotomy abolished the inhibition in one of the two cats but did not reduce the pressor response. Intrapericardial U-46619 injection did not affect the knee-jerk reflex nor blood pressure. The results indicate that U-46619 inhibited the knee-jerk reflex via a vagal reflex from the lung because the inhibition predominated after intravenous injection and was abolished by vagotomy. Speculation is made that the inflammatory mediator thromboxane A2 may contribute via a vagal reflex to the depression of motor activity associated with sickness behavior. PMID- 9728067 TI - Osmotic and freezing tolerance in spermatozoa of freeze-tolerant and -intolerant frogs. AB - The wood frog (Rana sylvatica) is a freeze-tolerant species that encounters subzero temperatures during its winter breeding season, whereas the leopard frog (R. pipiens) is freeze intolerant and breeds in spring. Osmotic and freezing tolerances of spermatozoa from these species were inferred from spermolysis rate, integrity of the plasma membrane as judged using vital dye assay, and motility rate. Sperm of R. sylvatica became motile in hypotonic media ( 5-carboxamidotryptamine (5-CT) >/= 8-hydroxy-2(di-n propylamino)tetraline (8-OH-DPAT); sumatriptan (Suma) produced no contractile response; and antagonist dissociation constant (pKb) values were 9.4 and 9.5 for ketanserin against 5-HT and 5-CT, 7.5 for GR-127935 against 5-HT, and 7.2 for SB 206553 against 5-HT. In normoxic adult Main segments, agonist potency order was 5 HT > 5-CT >/= Suma >/= DPAT, and pKb values were 9.1 and 9.2 for ketanserin against 5-HT and 5-CT and 7.4 and 8.5 for GR-127935 against 5-HT and Suma, respectively. In the 2BR segments from normoxic adults, agonist potency order was 5-CT > 5-HT > Suma > DPAT and pKb values were 7.4 and 7.2 for ketanserin against 5-HT and 5-CT and 10.0 and 8.7 for GR-127935 against 5-HT and Suma, respectively. Compared with normoxic adults, none of these values were significantly different in hypoxic adults and in fetuses only the pKb values for ketanserin against 5-HT in the 2BR segments (8.8) were greater. From these results we propose that the ratio of 5-HT2 to 5-HT1 receptors is greatest in the Com and decreases progressively to its smallest values in 2BR or smaller segments. Because this gradient appears stable and relatively resistant to the effects of maturation and chronic hypoxia, changes in reactivity associated with these perturbations may involve alterations in receptor density and/or coupling efficiency for 5-HT in ovine cranial arteries. PMID- 9728072 TI - Role of hypothalamic interleukin-1beta in fever induced by cecal ligation and puncture in rats. AB - Bacterial endotoxin induces fever by causing the release of interleukin (IL) 1beta into the circulation or the brain. IL-1beta is believed to mediate fever via triggering the production and/or release of IL-6 in the hypothalamus. The present study examined whether IL-1beta and IL-6 in the hypothalamus of the rat are also involved in fever during bacterial sepsis caused by cecal ligation and puncture (CLP). CLP induces fever for 2 days. Polyclonal rabbit antibody against rat IL-1beta (anti-IL-1beta, 2 microg/microl) or control rabbit IgG (2 microg/microl) was unilaterally microinjected into the hypothalamus of rats immediately after or 24 h after CLP or sham-CLP surgery. Anti-IL-1beta injected 24 h after CLP (when fever was already present) or sham-CLP surgery did not affect fever. Microinjection of anti-IL-1beta into the hypothalamus immediately after surgery caused a significant decrease in body temperature during the night after CLP surgery and a 48% reduction of fever on the following day. Although blood plasma levels of IL-6 were significantly elevated 1.5, 6, 24, and 48 h after CLP surgery, there were no differences in IL-6 concentrations in the extracellular fluid of the anterior hypothalamus (collected by push-pull perfusion). These data suggest that fever due to bacterial sepsis is initiated by IL-1beta within the hypothalamus, and this febrile response, unlike endotoxin induced fever, is not accompanied by elevation in the hypothalamic concentration of IL-6. PMID- 9728073 TI - Human duodenogastric reflux, retroperistalsis, and MMC. AB - The aim of this study was to determine to what extent human migrating motor complex (MMC)-related secretory phenomena are influenced by a recently discovered period of duodenal retroperistalsis during late phase III. A constant-flow perfusion technique was used to measure gastric appearance of acid, bicarbonate, pepsin, bilirubin, IgA, and duodenally infused [14C]polyethylene glycol (PEG) 4000 in 12 healthy volunteers. Interdigestive gastroduodenal motility was recorded by digital manometry. During late antral phase II and III, the gastric lumen was acidified (P < 0.005 phase III vs. phase I) together with a marked increase in luminal pepsin output (3.1 +/- 1.2 during phase III vs. 0.25 +/- 0.08 kU/5 min in phase I, P < 0.01), followed by a realkalinization due to a simultaneous reduction of acid secretion and a duodenogastric reflux, aided by retrograde peristalsis, of bicarbonate and IgA but not of bilirubin, at the end of antral phase III (P < 0.05 phase III vs. phase I values). This physiological duodenoantral reflux phenomenon may play an important role in the chemical and immunological restitution of the antral mucosal barrier function after the exposure to high acid and pepsin concentrations during antral phase III activity. PMID- 9728074 TI - Role of 17beta-estradiol in the modulation of baroreflex sensitivity in male rats. AB - Female mammals have an enhanced baroreflex sensitivity compared with their male counterparts, leading researchers to speculate that estrogen modulates autonomic tone. Therefore, this study tests the hypothesis that exogenous estrogen can enhance the baroreflex sensitivity of male rats. Male Sprague-Dawley rats anesthetized with thiobutabarbitol sodium (50 mg/kg) were instrumented to measure blood pressure and heart rate and for the intravenous injection of drugs. The baroreflex was tested using intravenous injections of phenylephrine (0.025, 0.05, and 0.1 mg/kg), and the cardiovascular responses were plotted to obtain a measure of the sensitivity of the cardiac baroreflex. Intravenous injection of estrogen produced dose-related increases in the baroreflex sensitivity due to an increase in the magnitude of the reflex bradycardia. In a separate group of animals, stimulation of the vagus nerve for 2 h resulted in a decrease in baroreflex sensitivity. This effect was blocked when estrogen (1 x 10(-2) mg/kg) was administered immediately before the end of stimulation. In conclusion, intravenous injection of estrogen in male rats significantly enhanced baroreflex sensitivity and blocked the attenuation in the baroreflex sensitivity observed after vagal stimulation. PMID- 9728075 TI - Ablation of posterior atrial ganglionated plexus potentiates sympathetic tachycardia to behavioral stress. AB - The role of the posterior atrial ganglionated plexus (PAGP) in heart rate (HR) control was tested in unanesthetized dogs (n = 8). Resting HR was unchanged before (85 +/- 20 beats/min, mean +/- SD) versus after (87 +/- 18 beats/min) surgical ablation of these intrinsic cardiac ganglia (PAGPX). However, the peak tachycardia to a 30-s stressful stimulus was significantly increased (P < 0.05) from +53 +/- 22 beats/min before the denervation to +77 +/- 13 beats/min after PAGPX. Conversely, the peak HR increase during the stress after beta-adrenergic blockade was the same before (36 +/- 24 beats/min) versus after (38 +/- 14 beats/min) PAGPX. Moreover, the HR response to a neutral behavioral stimulus, which is mediated primarily by withdrawal of parasympathetic inhibition of the sinoatrial (SA) node, was unaltered by PAGPX. Thus the augmented tachycardia subsequent to PAGPX was attributable primarily to increased sympathetic action at the SA node. These findings indicate that a major role of PAGP parasympathetic neurons is to inhibit sympathoexcitatory effects on HR, probably either via interactions between neurons comprising the intrinsic plexus(es) or perhaps via presynaptic inhibition of sympathetic neurotransmitter release. This organization would allow parasympathetic ganglia within the PAGP to selectively modify sympathetic input to the SA node independent of direct vagal inhibition of pacemaker activity. PMID- 9728076 TI - Endothelial dysfunction precedes hypertension in diet-induced insulin resistance. AB - The insulin-resistant (IR) syndrome may be an impetus for the development of hypertension (HTN). Unfortunately, the mechanism by which this could occur is unclear. Our laboratory and others have described impaired endothelium-mediated relaxation in IR, mildly hypertensive rats. The purpose of the current study is to determine if HTN is most likely a cause or result of impaired endothelial function. Sprague-Dawley rats were randomized to receive a fructose-rich diet for 3, 7, 10, 14, 18, or 28 days or were placed in a control group. The control group received rat chow. After diet treatment, animals were instrumented with arterial cannulas, and while awake and unrestrained, their blood pressure (BP) was measured. Subsequently, endothelium-mediated relaxation to acetylcholine was determined (in vitro) by measuring intraluminal diameter of phenylephrine preconstricted mesenteric arteries ( approximately 250 microM). Serum insulin levels were significantly elevated in all groups receiving fructose feeding compared with control, whereas there were no differences in serum glucose levels between groups. Impairment of endothelium-mediated relaxation starts by day 14 [mean percent maximal relaxation (Emax): 69 +/- 10% of baseline] and becomes significant by day 18 (Emax: 52 +/- 11% of baseline; P < 0.01). However, the mean BP (mmHg) does not become significantly elevated until day 28 [BP: 132 +/- 1 (day 28) vs. 116 +/- 3 (control); P < 0.05]. These findings demonstrate that both IR and endothelial dysfunction occur before HTN in this model and suggest that endothelial dysfunction may be a mechanism linking insulin resistance and essential HTN. PMID- 9728077 TI - Blockade of corticotropin-releasing hormone receptors reduces spontaneous waking in the rat. AB - We have previously hypothesized that corticotropin-releasing hormone (CRH) is involved in the regulation of physiological waking. To further elucidate this role for CRH, we administered intracerebroventricularly into rats two specific CRH-receptor antagonists, alpha-helical CRH-(9-41) (alpha-hCRH) or astressin, and determined changes in electroencephalogram-defined waking and sleep. Our results indicate that both of these receptor antagonists reduce the amount of time spent awake in a dose-related manner when administered before the dark period of the light-dark cycle. However, the time courses for these effects differ between antagonists; effective doses of alpha-hCRH reduce waking during the first 2 h postinjection, whereas effective doses of astressin reduce waking during postinjection hours 7-12. In contrast to dark-onset administrations, the amount of waking is not altered by either CRH-receptor antagonist when administered before the light period. These results support our hypothesis that CRH contributes to the regulation of physiological waking, since interfering with the binding of CRH to its receptor reduces spontaneous waking. PMID- 9728078 TI - IL-1beta increases norepinephrine level in rat frontal cortex: involvement of prostanoids, NO, and glutamate. AB - The effects of local administration of interleukin-1beta (IL-1beta) were studied by using an intracerebral microdialysis technique in rats. A local injection of IL-1beta (3 and 10 ng) induced an elevation of norepinephrine (NE) concentration in the medial prefrontal cortex (mPFC). IL-1-receptor antagonist (800 ng) completely blocked the IL-1beta-induced NE increase. Diclofenac, a cyclooxygenase inhibitor (500 microM), and Nomega-nitro-L-arginine, a nitric oxide (NO) synthase inhibitor (100 microM), applied through the dialysis probe, did not affect the initial rise in NE levels observed 20 min after injection of IL-1beta but completely suppressed the late phase of IL-1beta-induced NE increase at 40 min and thereafter. In contrast, local perfusion of 6-cyno-7-nitroquinoxaline-2,3 dione, a non-N-methyl-D-aspartic acid (NMDA) glutamate-receptor antagonist (50 microM), but not DL-2-amino-5-phosphonovaleric acid, an NMDA-receptor antagonist (100 microM), blocked both phases of IL-1beta-induced NE increase. Furthermore, a microinjection of IL-1beta elevated the extracellular concentration of glutamate in the mPFC. These findings suggest that the IL-1beta-induced rise in NE levels in the mPFC is caused by activation of the glutamatergic system and the glutamate induced increases in prostanoids and NO. PMID- 9728079 TI - Circadian rhythms of lipoprotein lipase and hepatic lipase activities in intermediate metabolism of adult rat. AB - Although intermediate metabolism is known to follow circadian rhythms, little information is available on the variations in lipoprotein lipase (LPL) and hepatic lipase (HL) activities during the 24-h period, and there is also a lack of adequate statistical analysis. Here, adult male rats were fed ad libitum and kept at 21 degrees C under 12:12-h light-dark cycles. They were killed in batches every 3 h over a 24-h period. Lipase activities were determined in plasma and fresh homogenates of epididymal white adipose tissue (EWAT), interscapular brown adipose tissue (IBAT), heart, skeletal muscle, and liver. Plasma insulin, corticosterone, glucose, triacylglycerol (TAG), cholesterol, glycerol, beta hydroxybutyrate, and liver and muscle glycogen were determined. Cosinor analysis was used to evaluate the presence (significance of fit of cosine curve to data and variance explained by rhythm) and characteristics of possible circadian rhythms [acrophase (phi), mesor, and amplitude]. Statistically significant circadian rhythms were detected for 1) all metabolites studied, except TAG, cholesterol, and liver HL activity; 2) LPL and HL activity in plasma (both phi in light phase); and 3) LPL activity in all tissues studied (phi: heart in light phase; skeletal muscle, IBAT, and EWAT in dark phase). Liver also showed a circadian rhythm of LPL activity, with phi near that in plasma. These findings demonstrate for the first time that, in physiological conditions, LPL activities in plasma and various tissues, including liver, and HL activity in plasma follow circadian rhythms. Their metabolic significance is discussed. PMID- 9728080 TI - No evidence for homeoviscous adaptation in a heterothermic tissue: tuna heat exchangers. AB - Many poikilotherms are known to adjust the membrane composition of their cells in response to a temperature change so that membrane fluidity, and therefore function, is conserved. Such compensatory changes in membrane composition are considered "homeoviscous adaptations." In this study, we examined a heterothermic tissue, the visceral rete mirabile of the bluefin tuna, for evidence of homeoviscous adaptation. We measured the proportions of phospholipid fatty acids and phospholipid head groups as a function of position along the rete thermal gradient, which has been estimated to be approximately 10 degrees C. We found no effect of position along the rete on the composition of either phospholipid fatty acids or head groups. Our results were unexpected in light of our previous demonstration of compensation of metabolic enzyme activity in the same tissue. The lack of evidence for a homeoviscous response may be due to the fluctuating nature of the thermal gradient along the visceral retia; i.e., membranes may be adapted to a eurythermal existence rather than being fine-tuned to a particular temperature. PMID- 9728081 TI - Regional and functional differences in the distribution of vestibulosympathetic reflexes. AB - Although considerable evidence suggests that the vestibular system regulates sympathetic outflow during movement and changes in posture, little is known about relative vestibular influences on activity of different sympathetic nerves and sympathetic efferents with different functions. In the present study, we demonstrated that electrical stimulation of the vestibular nerve in the cat elicited responses in sympathetic nerves innervating the head and abdominal viscera. This observation suggests that activity of sympathetic efferents innervating multiple body regions is affected by vestibular signals. These responses were attenuated by >80% when blood pressure was increased to >160 mmHg. Because raising blood pressure decreases the responsiveness of vasoconstrictor fibers, the simplest explanation for these data is that the vestibular system provides particularly strong inputs to components of the sympathetic nervous system that regulate peripheral vascular resistance. Furthermore, the relative magnitude of vestibulosympathetic reflexes was over four times larger in one sympathetic nerve composed mainly of vasoconstrictor efferents (renal nerve) than another nerve (external carotid nerve) containing similar types of fibers. Collectively, these data indicate that the vestibular system has selective influences on sympathetic outflow to particular tissues and body regions. PMID- 9728082 TI - Effect of ventilation style on cardiovascular and renal adaptation in preterm newborn lambs. AB - Renal adaptive responses during the 24 h after delivery in term newborn lambs include marked increases in both glomerular filtration rate (GFR) and sodium reabsorption. This study investigated the effects of ventilation style on cardiovascular, renal, and endocrine adaptations in preterm newborn lambs. Lambs (n = 62) were delivered by cesarean section at 131 days gestation (term = 150 days), treated with surfactant, and randomized to one of three ventilation strategies: high-frequency oscillation (12 Hz), high rate (50 breaths/min; tidal volume = 8 ml/kg), or low rate (15 breaths/min; tidal volume = 15 ml/kg). Lambs (5 or 6/group) were ventilated for 2, 5, 10, and 24 h to maintain arterial PCO2 between 45 and 50 mmHg. Plasma vasopressin levels decreased to <25 pg/ml by 10 h, and fractional sodium excretion decreased to <1% by 16 h in all groups. However, cardiac output, renal plasma flow, and GFR values did not change over time for any of the groups. The style of ventilation employed had no measurable effects on overall cardiovascular, renal, or endocrine function. We conclude in ventilated preterm lambs that 1) the ventilation style does not affect the time course for postnatal adaptation, 2) adaptive changes in renal tubular sodium reabsorption are evident by 16 h after birth, and 3) changes in preterm newborn renal sodium reabsorption occur in the absence of postnatal changes in renal plasma flow or GFR. PMID- 9728083 TI - Effect of heating on the hemodynamic responses to vasoactive agents. AB - During hyperthermia, vasoconstrictor tone in the viscera is lost despite high levels of sympathetic neural outflow and plasma catecholamines, suggesting that vascular responsiveness to adrenergic receptor stimulation is reduced. The purpose of this study was to determine whether adrenoceptor-mediated control of vascular resistance is altered at high body core temperatures. The hemodynamic responses to adrenoceptor agonists were examined in chloralose-anesthetized rats heated to colonic temperatures (Tco) of 37, 39, and 41.5 degrees C. Elevating Tco to 39 degrees C did not alter the hemodynamic responses to any of these agents. Further heating to 41.5 degrees C markedly attenuated the hemodynamic responses to alpha- and beta-adrenoceptor agonists. Similarly, the regional and systemic hemodynamic responses to ANG II and endothelin were also reduced at 41.5 degrees C. In contrast, the hemodynamic responses to endothelium-dependent and independent vasodilator agents were unchanged or slightly reduced at 41.5 degrees C. The blunted hemodynamic responses observed at 41.5 degrees C indicate that vascular reactivity to vasoconstrictor agents is reduced with hyperthermia and suggest that this nonspecific change in vascular responsiveness may contribute the circulatory collapse associated with high body temperatures. PMID- 9728084 TI - Evaluation of different levels of hydration using a new physiological strain index. AB - A physiological strain index (PSI), based on rectal temperature (Tre) and heart rate (HR), was recently suggested for evaluating heat stress. The purpose of this study was to evaluate the PSI for different combinations of hydration level and exercise intensity. This index was applied to two databases. The first database was obtained from eight endurance-trained men dehydrated to four different levels (1.1, 2.3, 3.4, and 4.2% of body wt) during 120 min of cycling at a power output of 62-67% maximum O2 consumption (VO2 max) in the heat [33 degrees C and 50% relative humidity (RH)]. The second database was obtained from nine men performing exercise in the heat (30 degrees C and 50% RH) for 50 min. These subjects completed a matrix of nine trials of exercise on a treadmill at three exercise intensities (25, 45, and 65% VO2 max) and three hydration levels (euhydration and hypohydration at 3 and 5% of body wt). Tre, HR, esophageal temperature (Tes), and local sweating rate were measured. PSI (obtained from either Tre or Tes) significantly (P < 0.05) differentiated among all exposures in both databases categorized by exercise intensity and hydration level, and we assessed the strain on a scale ranging from 0 to 10. Therefore, PSI applicability was extended for heat strain associated with hypohydration and continues to provide the potential to be universally accepted. PMID- 9728085 TI - Thermal acclimation of phase behavior in plasma membrane lipids of rainbow trout hepatocytes. AB - The fluorescent probes laurdan (6-dodecanoyl-2-dimethylaminonapthalene) and N-[7 nitrobenz-2-oxa-1, 3-diazol-4-yl] dipalmitoyl-L-alpha-phosphatidylethanolamine (NBD-PE) in addition to Fourier transform infrared spectroscopy (FTIR) were employed to measure the phase behavior and physical properties of hepatocyte plasma membranes isolated from the livers of thermally acclimated (5 and 20 degreesC) rainbow trout (Oncorhynchus mykiss). The primary objective was to determine the extent to which the phase behavior of membrane lipids is conserved at different growth temperatures. Arrhenius plots of laurdan-generalized polarization revealed a single discontinuity believed to reflect either the onset of the gel-fluid phase transition or the formation of gel phase microdomains, and this discontinuity occurred at significantly higher temperatures in membranes of 20 degrees C (13.2 +/- 0.7 degrees C)- than 5 degrees C (7.2 +/- 0.1 degrees C) acclimated trout. Similarly, acclimation from 5 to 20 degrees C increased both the onset temperature (from 2.0 +/- 0.3 to 7.2 +/- 0.6 degrees C) and the thermal range (from 10.9 +/- 0.5 to 16.0 +/- 1.0) of the gel-fluid transition as assessed by FTIR. The gel-fluid transition midpoint (approximately -2 degrees C) and completion temperatures (-9 degrees C) were unchanged by thermal acclimation. The anisotropy of NBD-PE fluorescence displayed a distinct minimum in membranes of both warm- and cold-acclimated trout (reflecting alterations in lipid packing that in pure lipid membranes ultimately lead to the formation of nonlamellar phases) in the range of 56-58 degrees C; only membranes of 5 degrees C-acclimated trout displayed an additional minimum at significantly lower temperatures (24.5 +/- 1.7 degrees C). Collectively, these data suggest that the regulation of both the temperature at which gel phase lipids begin to form in response to cooling as well as the propensity of membrane lipids to form nonlamellar phases at higher temperatures may be key features of membrane organization subject to adaptive regulation. PMID- 9728087 TI - Preventing hemodilution abolishes natriuresis of water immersion in humans. AB - The hypothesis was tested that hemodilution is one of the determinants of the water immersion (WI)-induced natriuresis. Eight males were subjected to 3 h of 1) WI to the midchest (Chest), 2) WI to the neck combined with thigh cuff-induced (80 mmHg) venous stasis (Neck + stasis), and 3) a seated time control (n = 6). Central venous pressure and left atrial diameter increased to the same extent during Chest and Neck + stasis (P < 0.05), whereas renal sodium excretion only increased during Chest from 77 +/- 7 to 225 +/- 13 micromol/min (P < 0.05). During Chest, plasma colloid osmotic pressure (COP) decreased from 27.7 +/- 0.7 to 25.1 +/- 0.7 mmHg (P < 0.05), and plasma volume (PV) increased from 3,263 +/- 129 to 3,581 +/- 159 ml (P < 0.05), whereas these variables remained unchanged during Neck + stasis. Plasma norepinephrine concentration decreased similarly during Chest and Neck + stasis by 45 +/- 7 and 34 +/- 4%, respectively (P < 0.05), whereas plasma renin activity decreased only during Chest (P < 0.05). In conclusion, during WI in humans 1) hemodilution (decrease in COP and increase in PV) is a pivotal stimulus for the natriuresis and 2) central blood volume expansion without hemodilution does not augment renal sodium output. PMID- 9728086 TI - Acute stressor exposure both suppresses acquired immunity and potentiates innate immunity. AB - Acute stressor exposure alters immune function. Rats exposed to inescapable tail shock stress (IS) generate less antibody to a benign, antigenic protein, keyhole limpet hemocyanin (KLH). The following studies examined the effect of IS on peritoneal cavity, spleen, and mesenteric lymph node cell number, interferon gamma (IFN-gamma) production, and nitrite production. Rats were injected intraperitoneally with KLH (200 microg) or saline immediately before IS exposure and killed 0, 48, and 96 h after IS termination. KLH immunization resulted in elevated cell numbers and IFN-gamma levels 2-4 days later in nonstressed control rats. In contrast, rats exposed to IS failed to increase cell number and IFN gamma levels in response to KLH. The T cell subpopulations affected were CD4 T cells, specifically the Th1-like subset. In addition, in rats exposed to IS + KLH, nitrite production was potentiated 2-4 days after stressor termination. IS had little effect on these measures in saline-injected rats. These data support the conclusion that exposure to IS suppresses the expansion of anti-KLH lymphocytes, possibly anti-KLH Th1 cells. In addition, stressor exposure potentiates the production of nitrite. Importantly, this potentiated response occurred only in KLH-immunized animals, suggesting that macrophages may be primed by stressor exposure and thus respond more vigorously to antigen. The potential links between these changes are discussed. PMID- 9728088 TI - Raised intracranial pressure increases CSF drainage through arachnoid villi and extracranial lymphatics. AB - We demonstrated previously that about one-half of cerebrospinal fluid (CSF) removed from the cranial vault was cleared by extracranial lymphatic vessels. In this report we test the hypothesis that lymphatic drainage of CSF increases as intracranial pressure (ICP) is elevated in anesthetized sheep. Catheters were inserted into both lateral ventricles, cisterna magna, cervical lymphatics, and jugular vein. A ventriculocisternal perfusion system was employed to regulate CSF pressures and to deliver a protein tracer (125I-labeled human serum albumin) into the CSF compartment. 131I-labeled human serum albumin was injected intravenously to permit calculation of plasma tracer loss and tracer recirculation into lymphatics. ICP was controlled by adjusting the height of the inflow reservoir and the cisterna magna outflow catheter appropriately. The experimental design consisted of a 3-h period of lower pressure followed by a 3-h period of higher pressure in the same animal (10-20 or 20-30 cmH2O). We determined that incremental changes in ICP were associated with higher CSF transport through lymphatic and arachnoid villi routes in all eight animals tested (P = 0.004). PMID- 9728089 TI - Ontogenetic and regional changes in alpha-methyl-D-glucoside and L-proline intestinal transport in guinea pig. AB - Regional brush-border uptakes of alpha-methyl-D-glucose and L-proline as well as morphometric parameters were studied from birth until adulthood in guinea pig small intestine. Intestinal weight, length, and area were fitted to two-segmented straight lines: from birth until the 2nd wk there was a sharp rise, whereas from day 14 to the adult stage the increase was slower. In everted sleeves, total uptakes of alpha-methyl-D-glucoside were higher on day 1 in duodenum, jejunum, and ileum, diminishing with increasing age. The initial fluxes of L-proline were higher during the 1st wk, diminishing to values that kept constant thereafter. Total uptakes of L-proline relative to alpha-methyl-D-glucoside showed a peak in the 1st wk in the three segments studied, reflecting the high demand for protein during the postnatal period. Regional ratios of L-proline to alpha-methyl-D glucoside indicated that the ileum is the segment best suited to transporting this amino acid during the first 3 wk. Changes observed in the present study indicated a different pattern between hexose and amino acid transport during development and along the small intestine of guinea pig. PMID- 9728090 TI - Influence of acclimation temperature on mitochondrial DNA, RNA, and enzymes in skeletal muscle. AB - Skeletal muscle fibers typically undergo modifications in their mitochondrial content, concomitant with alterations in oxidative metabolism that occur during the development of muscle fiber and in response to physiological stimuli. We examined how cold acclimation affects the mitochondrial properties of two fish skeletal muscle fiber types and how the regulators of mitochondrial content differed between tissues. After 2 mo of acclimation to either 4 or 18 degrees C, mitochondrial enzyme activities in both red and white muscle were higher in cold acclimated fish. No significant differences were detected between acclimation temperatures in the abundance of steady-state mitochondrial mRNA (cytochrome-c oxidase 1, subunit 6 of F0F1-ATPase), rRNA (16S), or DNA copy number. Steady state mRNA for nuclear-encoded respiratory (adenine nucleotide translocase 1) and glycolytic genes showed high interindividual variability, particularly in the cold-acclimated fish. Although mitochondrial enzymes were 10-fold different between the two muscle types, mitochondrial DNA copy number differed only 4-fold. The relative abundance of mitochondrial mRNA and nuclear mRNA in red and white muscle reflected the differences in copy number of their respective genes. These data suggest that the response to physiological stimuli and determination of tissue-specific mitochondrial properties likely result from the regulation of nuclear-encoded genes. PMID- 9728091 TI - Leptin causes body weight loss in the absence of in vivo activities typical of cytokines of the IL-6 family. AB - To investigate if leptin shares in vivo activities with interleukin (IL)-6 family cytokines, it was tested in normal mice for the ability, after a single injection, to induce the acute-phase protein serum amyloid A, to potentiate the induction by IL-1 of serum corticosterone and IL-6, and to inhibit the induction by lipopolysaccharide of serum tumor necrosis factor and, after seven daily injections, to cause body weight loss and to change peripheral blood cell counts. At a 0.5 mg/kg dose, leptin caused body weight loss but did not show any of the other activities above. At a dose of 5 mg/kg, which also caused body weight loss, leptin potentiated the induction by IL-1 of serum corticosterone and IL-6 but did not show any other activity. In addition to causing body weight loss, leptin shows only some of the in vivo activities typical of IL-6 family cytokines and only if used at a dose that exceeds the one sufficient to affect body weight. In vivo, leptin seems to chiefly control body weight and not inflammatory or hematopoietic processes. PMID- 9728092 TI - Mechanisms of spontaneous baroreflex impairment in lyon hypertensive rats. AB - This experiment aimed at 1) comparing the spontaneous baroreflex sensitivity (SBRS) in Lyon genetically hypertensive (LH), normotensive (LN), and low blood pressure (LL) rats and 2) assessing some aspects of the mechanisms of its impairment in LH rats. Baroreflex was studied in control animals after an early chronic converting enzyme inhibition with perindopril and after a 4-wk infusion of ANG II in perindopril-treated rats. The SBRS was determined with a previously validated method, using statistical dependence between blood pressure (BP) and heart rate values recorded in freely moving animals. LH rats exhibited high BP, cardiac hypertrophy, and decreased SBRS (LH, 1.3 +/- 0.2; LN, 2.5 +/- 0.4; LL, 2.2 +/- 0.4 beats . min-1 . mmHg-1). Perindopril prevented the development of hypertension and cardiac hypertrophy and normalized SBRS. BP rose in LH and LL rats after ANG II infusion, but only LH rats, which developed a cardiac hypertrophy, had an impaired SBRS (LH, 1.1 +/- 0.2; LN, 2.5 +/- 0.2; LL, 2.8 +/- 0.3 beats . min-1 . mmHg-1). This impairment was partially reversed by an acute ANG II blockade with losartan. These results demonstrate that high BP does not account for the decreased SBRS in LH rats. SBRS impairment could result either from cardiac hypertrophy or from the direct effect of ANG II on the baroreflex loop. PMID- 9728093 TI - Isoform expression of Na+-K+-ATPase alpha-subunit in gills of the teleost Oreochromis mossambicus. AB - Three isoform-specific antibodies, 6F against the alpha1-isoform of the avian sodium pump, HERED against the rat alpha2-isoform, and Ax2 against the rat alpha3 isoform, were used to detect the expression of Na+-K+-ATPase alpha-subunits in gills of a teleost, the tilapia (Oreochromis mossambicus). Tilapia gill tissue showed positive reactions to antibodies specific for alpha1- and alpha3-isoforms. The results of immunoblots were converted to numerical values (relative intensities) by image analysis for comparisons. Relative amounts of alpha1-like isoform alone and consequently the ratio of alpha1-like to alpha3-like isoforms were higher in gills of seawater-adapted tilapia than in those of freshwater adapted ones, indicating that the two isoforms respond differently to environmental salinities. In the subsequent immunocytochemical experiments, gill mitochondria-rich cells were demonstrated to immunoreact with antibodies specific for alpha1- and alpha3-isoforms. alpha1-like and alpha3-like isoforms of gill Na+ K+-ATPase are suggested to be involved in the ion- and osmoregulation mechanisms in tilapia. Moreover, differential expressions of two isoforms may be associated with different functions, secretion and uptake of ions and acid-base regulation, in gills of seawater- and freshwater-adapted tilapia. PMID- 9728094 TI - Investigation of nitrous oxide pollution arising from inhalational sedation for the extraction of teeth in child patients. AB - OBJECTIVES: (i) TO test whether the exposure of dental staff to nitrous oxide during inhalational sedation with nitrous oxide/oxygen for extractions in children complies with specified occupational exposure standards, and (ii) to assess the atmospheric nitrous oxide concentration at one site close to the breathing zone of the operator/sedationist and to determine which patient- and sedation-related factors affect the level of nitrous oxide pollution. DESIGN: Prospective study. SETTING: Dental hospital sedation department, Newcastle Dental Hospital, UK. SAMPLE AND METHODS: 20 inhalational sedation clinics each of 2 hours duration were evaluated, during which a total of 60 children aged 4-15 years had extractions carried out. Nitrous oxide was administered via a nose mask from a Quantiflex MDM inhalational sedation machine and active scavenging was used throughout. Exposure of dental staff was measured using personal dosimetry. Atmospheric nitrous oxide pollution at one fixed point, close to the breathing zone of the operator/sedationist, was assessed using infra-red gas analyser. RESULTS: Mean exposure of the operator/sedationist to nitrous oxide during a single treatment clinic was 211 ppm, for the close support nurse 77 ppm and for the second nurse 67 ppm. Expressed as an 8 hour time-weighted average, the mean exposures were 39 ppm for the operator/sedationist, 17 ppm for the close support nurse and 15 ppm for the second nurse. The atmospheric nitrous oxide concentration varied during the clinics, with a maximum concentration of 538 ppm an a minimum that exceeded 100 ppm. There was a 71 minute delay following discharge of the last patient before atmospheric levels fell to zero. During sessions the degree of atmospheric pollution was inversely related to patient age (rc = -0.61, P < 0.05). There was a positive correlation between atmospheric pollution at the single point and the maximum percentage of nitrous oxide administered to each patient (rc = 0.57, P < 0.05). CONCLUSIONS: In this study, staff exposure to nitrous oxide complied with the national occupational exposure standard but there was still considerable atmospheric nitrous oxide pollution during inhalational sedation for paediatric exodontia. PMID- 9728095 TI - Bond strength and interfacial micromorphology of compomers in primary and permanent teeth. AB - OBJECTIVES: To (1) test and compare the shear bond strength of compomers (Compoglass, Dyract, Hytac) to primary and permanent dentin, (2) compare the values to those obtained with a resin-modified glass ionomer (Vitremer), and (3) evaluate the material-dentin interfacial morphology. SAMPLE AND METHODS: The facial and lingual surfaces of 32 primary and 32 permanent teeth were used. The manufacturers' instructions were followed for the bonding procedures. After bonding, the teeth were thermocycled and sheared. RESULTS: ANOVA and Student Newman-Keuls test revealed that the shear bond strength for Dyract was significantly higher than for the other restorative systems tested, both for primary (P < 0.001) and permanent (P < 0.01) teeth. Compoglass bond strength was significantly lower than Vitremer for the primary teeth dentin (P < 0.01). The shear bond strength for Compoglass to permanent dentin was significantly lower than for all other restorative systems (P < 0.001). There was a significantly higher shear bond strength for Compoglass (P < 0.05) and Dyract (P < 0.01) restorative systems for primary compared to permanent teeth. For all products tested, all samples revealed cohesive failures. The highest frequency of cohesive failure was reported with Compoglass in both primary and permanent teeth and for Hytac and Vitremer in permanent teeth. Micromorphologically, all restorative systems revealed good adaptation to the underlying dentin; however, there was no evidence of the formation of a hybrid layer or deep resin penetration inside the dentinal tubules. There was no difference in the interfacial morphological adaptation between the primary and permanent teeth. CONCLUSIONS: The compomers tested had shear bond strength values between those of resin-modified glass ionomers and resin composites. PMID- 9728096 TI - Caries prediction model in pre-school children in Riyadh, Saudi Arabia. AB - OBJECTIVES: To evaluate the significance of variables such as oral hygiene, dietary habits, socio-economic status and medical history of a child in assessing the level of caries risk and to generate a caries prediction model for pre-school Saudi children. DESIGN: Cross-sectional study of pre-school children. SETTING: Clinics and schools in Riyadh, Saudi Arabia. SAMPLE AND METHODS: A sample of 446 Saudi pre-school children, 199 males and 247 females, with a mean age of 4.13 years, were selected at random from clinics and schools. Selection was limited to subjects who either had no caries (dmft = 0) or who had high caries experience (dmft > 8). Each child was examined for caries experience and oral hygiene status. Their mothers were interviewed through a standardized questionnaire for information about oral hygiene habits of the children, diet history, childhood illness and socio-economic status. RESULTS: There was a highly significant difference between the two groups in: debris index (P < 0.0001), aged child started tooth brushing, (P < 0.0001), age breastfeeding was stopped (P < 0.005), nocturnal bottle feeding with milk formula (P < 0.001), use of sweetened milk (P < 0.0001), frequency of use of soft drinks (P < 0.0005), frequency of consumption of sweets (P < 0.0001), and age at first dental visit (P < 0.0001). A caries prediction model developed through stepwise multivariate Logistic Regression (LR) analyses showed debris index, use of sweetened milk in bottle, frequency of consumption of soft drinks, frequency of intake of sweets and child's age at the first dental visit to be significant. Predictive probability of the model was 86.31% with a sensitivity of 90.1% and a specificity of 80.6%. CONCLUSIONS: Risk factors for dental caries have been identified and a caries prediction model has been developed for Saudi pre-school children. The prediction model, if verified, may provide with guidance in identifying high caries risk Saudi preschool children as targets for preventive programmes. PMID- 9728097 TI - The effect of two alternative methods of canine exposure upon subsequent duration of orthodontic treatment. AB - OBJECTIVES: The aim of this study was to compare the effectiveness, in terms of orthodontic treatment duration, of two methods of canine exposure. DESIGN: This was a retrospective study using patients' records and lateral cephalometric radiographs. SAMPLE AND METHODS: 50 patients were selected, 25 in each group. In all subjects the impaction was categorized as being 'intermediate'. The methods of canine exposure were: (i) simple surgical exposure; (ii) surgical exposure and placement of an orthodontic attachment, followed by flap replacement. RESULTS: The treatment duration until the canine was in the line of the arch was 17.7 months in the simple exposure group and 19.3 months in the bonded attachment group. The mean treatment duration (from exposure to debond) was 28.8 months for both groups. CONCLUSIONS: In terms of treatment duration, no significant difference could be demonstrated between the two methods of surgical exposure to palatally impacted canines. PMID- 9728098 TI - Demographic characteristics, oral health knowledge and practices of mothers of children aged 5 years and under referred for extraction of teeth under general anaesthesia. AB - AIMS: To examine the demographic characteristics, oral health knowledge and practices of mothers of children who required the extraction of teeth under general anaesthesia. DESIGN: Cross-sectional study using questionnaires completed at interview. SAMPLE: 150 mothers of children age 5 years and under who had been referred for extraction of teeth under general anaesthesia. SETTING: Dental School, University of Wales College of Medicine. RESULTS: Over three-quarters of mothers had finished their full-time education at or before the age of 16 years. Although generally well-informed about the causes of caries, mothers were less sure of preventative methods. There was a widespread ignorance of the causes and prevention of periodontal disease, with just under half of the sample being unable to identify any causative or preventive factors. 40% of the sample admitted to attending a dentist only when in pain or not at all. PMID- 9728099 TI - Malocclusion, dental injuries and dental anomalies in the primary dentition of Belgian children. AB - AIM: To estimate the prevalence of malocclusion, dental injuries and dental anomalies in a sample of 3-5-year-old Belgian children. DESIGN: A cross-sectional study of 3-5-year-olds attending kindergartens in the municipality of Leuven, Belgium. METHODS: A total of 750 boys and girls participated in the study. The children were examined at the University School Health Centre in connection with their obligatory medical check-up. The clinical examination was performed by one examiner using generally accepted criteria for these oral conditions. RESULTS: 10.1% of the examined children had posterior cross-bite whereas over-bite was seen in only 2.0% of the sample. Open-bite was detected in 32.0% of the studied population. Boys showed a tendency for a higher frequency of malocclusions than girls. Traumatic injuries were identified in 18.0% of children. These were almost entirely restricted to maxillary incisors. Crown fractures were responsible for 42% of all injuries. The following dental anomalies were seen: six cases of supernumerary teeth, three cases of hypodontia, five cases of double teeth and one case of conical maxillary lateral incisor. CONCLUSION: Our findings emphasize the importance of early detection of these oral conditions in order to permit effective and long-term planning, according to the child's individual requirements. PMID- 9728100 TI - Traumatic injuries to the chin: a survey in a paediatric dental practice. AB - OBJECTIVE: The aim of the study was to provide data on the prevalence of clinical signs of traumatic injuries to the chin and of parental report of such injuries in a group of children visiting a private paediatric dental practice. SAMPLE AND METHODS: The study group comprised 303 children (151 boys and 152 girls) whose dental age was within the limits in which all primary molars are present. The children visited the author's private practice for purposes other than chin injury and its dental implications. Data collected included age, gender, presence and location of fractures in primary molars, presence of a scar on the child's chin, and parental report of previous chin injuries. RESULTS: Fractured primary molars were detected in 96 (31.7%) children. 79 (26%) children had a scar on the chin and previous chin injuries were reported for 110 (36.3%) children. Boys presented a higher prevalence of evidence of chin injuries than girls. The male/female ratios for fractured molars, scar on the chin and previous trauma were 1.5:1, 1.8:1 and 1.5:1 respectively. These differences were statistically significant (chi 2, P = 0.012, P = 0.002 and P = 0.008 respectively). CONCLUSIONS: It is concluded from this survey that traumatic injuries to the chin are not infrequent and the incidence of injuries to the chin is higher in boys than in girls. PMID- 9728101 TI - Multiple salivary mucoceles in a young boy. AB - A rare case of multiple mucoceles in a 4-year-old boy is reported. The patients presented with multiple sessile swellings on his lower lip, with no obvious local aetiology. The patient was otherwise well. The lesions were excised surgically and histopathological examination confirmed the lesions to be extravasation-type mucoceles. The article includes a brief review of mucoceles. PMID- 9728102 TI - Oral and dental anomalies in Ellis van Creveld syndrome (chondroectodermal dysplasia): report of a case. AB - A case of Ellis van Creveld syndrome is described in which, in addition to all the major features constituting the tetrad of chondroectodermal dysplasia, there was (unusually) bipedal hexadactyly. As well as the frequently reported oral anomalies, this case exhibited taurodontism of permanent and some primary molars, a finding rarely reported in this syndrome. PMID- 9728103 TI - Fissure sealants in a group of 3-4-year-old children. PMID- 9728104 TI - Six-year follow-up with Empress veneers. AB - This study reports on 6 years experience with IPS Empress laminate veneers. A total of 83 anterior veneers were positioned in 21 patients from January 1991 to December 1996 in the author's private practice. Final evaluation was carried out in May and June 1997. Color match, marginal discoloration, recurrent caries, contour, and marginal integrity were evaluated using the modified U.S. Public Health Service criteria at baseline and subsequent recall appointments. On the basis of the criteria used, a large percentage of veneers were rated Alfa. Only one failure was recorded, resulting in a success rate of 98.8%. A thorough description of clinical procedures and laboratory techniques through which anterior teeth can be successfully treated with ceramic veneers is supplied. A clinical case is presented to demonstrate the satisfactory esthetic results obtained using this very conservative restorative technique. PMID- 9728105 TI - Residual lateral wall defects following sinus grafting with recombinant human osteogenic protein-1 or Bio-Oss in the chimpanzee. AB - Sinus grafting procedures are a viable means of ensuring adequate bone for the placement of dental implants in the posterior maxilla. In the quest to improve predictability and accelerate the time line toward receiving a final prosthesis, researchers have turned to recombinant human proteins like osteogenic protein-1 for the potential to therapeutically enhance bone formation. Bilateral sinus augmentations were performed in 15 adults chimpanzees to evaluate treatment with different doses of the osteogenic protein-1 device or natural bone mineral (Bio Oss). Methods of evaluation included soft tissue healing, radiography (computed tomographic scan), histology, residual lateral wall defect surface area at 7.5 months, and the extent of soft tissue encleftation at 7.5 months. Findings revealed radiographic and histologic evidence of bone formation with all treatment groups and a statistically significant reduction in the depth of soft tissue encleftation and the residual lateral wall defect surface area for both the Bio-Oss and the 2.5-mg osteogenic protein-1 per gram collagen matrix treatments when compared to collagen matrix alone. These results suggest that Bio Oss and the 2.5-mg osteogenic protein-1 per gram collagen matrix effectively stimulate bone formation in the maxillary sinus. PMID- 9728106 TI - An orthodontic device to capitalize on the rigid fixation of osseointegrated implants. AB - Extraction of maxillary and mandibular first molars frequently results in tipping of the terminal molars. Ideal treatment involves uprighting the inclined teeth prior to restoration. If a single-tooth implant restoration will be used to replace the missing molar, it can be used as anchorage to upright the tipped or drifted teeth. Careful planning is necessary to make sure the implant is positioned properly for an ideal final result. PMID- 9728107 TI - A technique report on the in situ application of Atrisorb as a barrier for combination therapy. AB - This article describes a novel technique in which Atrisorb (Atrix Laboratories) is applied in situ as a barrier over a demineralized freeze-dried bone allograft on roots treated with citric acid. Follow-up reentry at 4.5 months demonstrated the effectiveness of this procedure. A rationale for in situ barrier application is presented. PMID- 9728108 TI - A new classification of molar furcation involvement based on the root trunk and horizontal and vertical bone loss. AB - This study presents a new classification of molar furcation involvement based on the root trunk and horizontal and vertical attachments, and relates them to guidelines in diagnosis, prognosis, and treatment of molar furcation involvements. The type of root trunk were classified on the basis of the ratio of vertical dimension of root trunk to root length as types A, B, and C. Based on the results of this study, the authors conclude that consideration of the root trunk type as well as the horizontal and vertical attachment loss in the classification of molar furcation involvements may facilitate prognosis, diagnosis, and treatment planning. PMID- 9728109 TI - Implant placement in large edentulous ridges expanded by GBR using a bioresorbable collagen membrane. AB - The purpose of this study is to evaluate the possibility of expanding an edentulous ridge spanning two or more teeth by a two-step technique with bioresorbable collagen membranes. Sixteen healthy patients were treated, four in the mandible and 12 in the maxilla. The borderline of the crest width was less than or equal to 4 mm. In each patient only one ridge augmentation was performed. After elevation of a full-thickness flap, a surgical stent was positioned to identify the area of ideal implant positioning. The width of the crest at the location of the surgical stent was measured at the time of GBR procedure and 7 to 12 months later during implant insertion. Native collagen sponges were placed buccally and lingually, and a collagen membrane was shaped and trimmed to completely cover the edentulous ridge. The flaps were sutured to achieve primary closure. Antibiotic and clorhexidine mouthrinse were prescribed, and the patients were recalled every 2 weeks. At implant placement, the mean increase in the size of the crest was 2.49 mm (+/- 1.61 mm). In 12 out of 16 patients (75%) it was possible to insert 27 implants according to the prosthetic need established previously. All implants were successfully loaded, and in the four cases where no appreciable results were obtained, no clinical complications or loss of hard and soft tissue were observed. PMID- 9728110 TI - The evolution of temporary fixed splints--the A-splint. AB - This article presents a review of the evaluation of the temporary splint leading up to the present-day acid-etched composite, wire-reinforced, internal splint called the "A-splint." Nine general uses for the A-splint in the modern dental practice and some common problems are presented. PMID- 9728111 TI - Soft tissue ridge augmentation to correct an esthetic deformity caused by adversely placed implants: a case report. AB - A patient presented with two osseointegrated implants placed in a ridge deformity that had resulted from a traumatic injury during an automobile accident. The implant placement and subsequent restorative dentistry resulted in a poor esthetic result and difficult maintenance. Treatment included multiple soft tissue grafts to submerge the implants and augment the ridge. The patient was then restored with a conventional fixed partial denture. PMID- 9728112 TI - Squamous cell carcinoma of gingiva and edentulous alveolar ridge: a clinicopathologic study. AB - All oral squamous cell carcinomas were retrieved from the files of Temple University's Oral Pathology Laboratory from 1967 through 1994 for a clinicopathologic study of those occurring on the gingiva. A total of 1,193 cases had sufficient data for tabulation and statistical analysis, of which 300 (25%) arose on the gingiva or alveolar ridge. The largest number of these cases (211/300) occurred on the mandibular gingiva or alveolar ridge. The mean age of the patients was 66.66 years, with males accounting for 57% of cases. Many case comparison analyses of oral squamous cell carcinomas do not separate oral subsites or specifically address carcinoma of the gingiva. The results were compared with other published series and suggest that further studies are needed because of the wide range of reported figures on the incidence of gingival squamous cell carcinomas. PMID- 9728114 TI - Cemented single crowns on osseointegrated implants after 5 years: results from a prospective study on CeraOne. AB - PURPOSE: The aim of this prospective study was to present the results after 5 years of loading of 65 CeraOne (Nobel Biocare) crowns. MATERIALS AND METHODS: Sixty-two implants in the maxilla and 3 implants in the mandible were placed in 57 patients. Sixty-two all-ceramic and three metal-ceramic crowns were cemented. The group comprised the first patients treated with the CeraOne prosthodontic concept. RESULTS: Eight patients did not complete the study. Only one implant failed, giving a cumulative success rate for implants of 98.5%. The failed implant was replaced: a crown was cemented and then followed for 5 years without any complications. Four crowns were recorded as failures, giving a cumulative success rate for crowns of 93.7%. It should be observed that this result was very positive, as all crown failures were related to extraordinary causes and not one was a result of common bite forces or fatigue. The initial bone loss was in accordance with other studies on Branemark implants, and a stable situation was recorded after 2 years for the supporting bone around implants and adjacent teeth when the conical implants were excluded. Soft tissues around implants and adjacent teeth appeared healthy, and the cementation and the placement of the abutment shoulder in the peri-implant sulcus did not cause any recession of the peri-implant mucosa. CONCLUSION: CeraOne experienced virtually no complications and proved to be a highly predictable and safe prosthodontic concept. CeraOne also eliminated problems with abutment screw loosening and created a platform for good esthetic results and satisfied patients. PMID- 9728113 TI - Improvement in the tensile bond strength between resin cement and dentin surfaces after temporary cement application. AB - PURPOSE: The aim of this study was to evaluate the efficacy of three tooth conditioners in restoring the reduced bond strength between resin cement and teeth resulting from remaining temporary cement. MATERIALS AND METHODS: Ethyl dihydrogen phosphate, methacryloxyethyl dihydrogen phosphate, and 2 methacryloxyethyl hydrogen maleate were evaluated as conditioners. After eliminating the temporary cement with a curette from the bovine dentin surface, the conditioners were applied onto the surface of the specimen and a resin cement was adhered. Stepwise scanning electron microscopic observation and tensile bond strength measurement were carried out. RESULTS: Granular substances were present on the dentin surface even when the temporary cement was carefully eliminated with a curette. When primer and resin cement were applied on this surface without conditioner application, no resin tag or hybrid layer was observed and the mean bonding strength between tooth and adhesive resin cement was 1.8 MPa. In contrast, resin tag and hybrid layer were observed after primer and resin cement application when the dentinal surface was treated with conditioner. Mean tensile bone strength values increased to 6.2 MPa for specimens treated with 20% methacryloxyethyl dihydrogen phosphate for 60 seconds. CONCLUSION: The authors recommend methacryloxyethyl dihydrogen phosphate as it provides high tensile bond strength values and requires no additional rinse step. PMID- 9728115 TI - Clinically relevant mechanical properties of elastomeric impression materials. AB - PURPOSE: This study investigates the modulus of elasticity, yield strength, the strain at yield point, and the tear energy of nine elastomeric impression materials. MATERIALS AND METHODS: The values of the first three variables were computed from a tensile load test of 10 dumbbell-shaped specimens of each impression material. Tear energy was calculated from the results of a standard trousers tear test on 10 specimens of each impression material. RESULTS: A general descending order of modulus of elasticity (rigidity) follows: poly(vinyl siloxane) putty > polyether > polysulfides and the poly(vinyl siloxane) tray and syringeable materials. The descending order of yield strength was: poly(vinyl siloxane) putty > polyether and most poly(vinyl siloxane) tray and syringeable materials > one poly(vinyl siloxane) and the two polysulfides. The general descending order in strain at yield point (strain tolerance) was: two poly(vinyl siloxane) syringeable materials > four poly(vinyl siloxane) materials of various viscosities > polyether and the two polysulfides. Tear energy followed a general descending order of: polysulfides > polyether > poly(vinyl siloxane). CONCLUSION: The difficulty of removing impressions made of the putty or the polyether, and the increased risk of die breakage could be associated with the higher rigidity of these materials. The high strain tolerance of the poly(vinyl siloxane) impression materials allows their removal without distortion from appreciable tissue undercuts. The high tear energy of polysulfides indicates their superiority over other impression materials in their resistance to tear in thin sections. PMID- 9728116 TI - Two years of clinical experience with Procera titanium crowns. AB - PURPOSE: The purpose of the study were to evaluate the clinical performance of the Procera porcelain-fused-to-titanium crown system in general practice during a 2-year period, and to evaluate the performance of a new low-fusing porcelain as a veneering material on titanium. MATERIALS AND METHODS: A number of consecutive complete-coverage crowns (40) in 25 patients (14 women and 11 men) with a mean age of 53.1 years (range 35 to 79 years) were made according to the Procera (Nobel Biocare) technique in a 3-month period. The titanium copings were fabricated from solid rods of pure titanium using spark erosion and copy-milling technique, whereafter they were veneered with a new type of low-fusing ceramic material. The crowns were evaluated using the CDA criteria at baseline and after 2 years. RESULTS: The general failure rate was low and was restricted to one carious lesion, one porcelain fracture, and one loss of a crown resulting from failure of retention of a post and core. The most frequent single reason that the "excellent" color level was not recorded was a "too-high value." A slightly dull or granular porcelain surface was observed both at baseline and after 2 years. Overall, the responses of the patients were positive. CONCLUSION: Within the limitations of the study it can be concluded that porcelain-veneered Procera titanium crowns can be used as an alternative to other porcelain-fused-to-metal systems. However, conclusions should be made with caution from the results of this study because of the limited number of patients and crowns and the short observation period. PMID- 9728117 TI - Tolerance test of five different types of crowns on single-tooth implants. AB - PURPOSE: The main purpose of the present experimental study was to compare five different types of crowns, cemented on implant abutments, regarding their capability to withstand loads. MATERIALS AND METHODS: Three types of all-ceramic crowns, a gold-foil-reinforced porcelain crown, and, as a control, a conventional metal ceramic crown were tested. Each crown was cemented onto an Astra Tech Single-Tooth implant. The five types of crowns, three of each type, and the titanium implants were subjected to loading in Lloyd test equipment until part of them was damaged, at which point the compression value was recorded and the deflection and bending moment were calculated. Comparisons were made on the basis of these data. RESULTS: The results showed that the all-ceramic crowns fitted with a core should be able to withstand normally occurring biting forces without difficulty. The foil crown was also judged to be acceptable, while the bending moment of the cast call-ceramic crown without a core was considered unpredictable. The values for the metal ceramic design were as predicted, ie, they were clearly the highest in the study; the superior strength of metal ceramics should still be taken into account when deciding between all-ceramic solutions and the conventional metal ceramic crown. CONCLUSION: It was concluded that all-ceramic crowns are weaker than conventional metal ceramic crowns: however, based on estimated maximum clinical loading (370 N in the incisor and premolar regions), In-Cream and AllCeram crowns seem to function satisfactorily on implants. PMID- 9728118 TI - Perceptions of complete dentures by prospective implant patients. AB - PURPOSE: The purpose of this study was to facilitate the recognition of denture patients who are unable to adapt to conventional dentures and who are likely to benefit from treatment using implant-supported prostheses. MATERIALS AND METHODS: Sixty-nine patients who where referred for postgraduate prosthetic treatment at Wits Dental School completed a self-report inventory of items related to their dentures in current use. Conventional dentures were fabricated for all subjects. Those patients who could not adapt to conventional complete denture treatment were referred for treatment with implant-supported prostheses provided that they conformed with the recommended criteria for this treatment modality. RESULTS: Analysis of the inventory of pretreatment denture complaints yielded variables that differentiated between the group who remained with conventional dentures and the group that was referred for implants. Significant variables were the period that a mandibular denture was used before new dentures were requested (P = 0.025); the period that a maxillary denture was used before further treatment was sought (P = 0.03); the discarding of a mandibular denture (P = 0.035); and pain complaints related to maxillary dentures (P = 0.045). A logistic regression model was used to compare the clinical division of the sample with that determined by the mathematic model. Sixty-six percent of the subjects who accepted conventional treatment and 69% of implant patients corresponded for both classifications. CONCLUSION: The authors conclude that pretreatment denture complaints can be used diagnostic aids for evaluating patients who are likely to benefit from implant supported prostheses. PMID- 9728119 TI - An international comparative multicenter study of assessment of dental appearance using computer-aided image manipulation. AB - PURPOSE: The aim of the present investigation was to perform an international multicenter comparison of dental appearance as evaluated by dentists, dental technicians, and nondental subjects. MATERIALS AND METHODS: The participants were drawn from three groups: 203 dentists, 197 dental technicians and 254 nondental subjects. The methods developed in a previous study in Sweden were applied again in seven centers located in six countries. A questionnaire, accompanied by five sets of computer-manipulated images portraying one man and one woman, was used to prompt and record responses to different aspects of dental appearance and function. RESULTS: The questionnaire revealed that both the dental appearance and function of teeth were important to most of the participants, but three quarters of the participants did indicate that good dental function was more important that esthetics. More women (30%) than men (18%), however, placed greater importance on appearance. Age or gender did not influence judgments of the computer-manipulated images, although judgments did vary greatly within the three groups and between the centers. Nonetheless, highly colored teeth were preferred more often by nondental subjects than by dentists or dental technicians. CONCLUSION: Computer-aided image manipulation shows promise as a method for investigating the significance of dental-related beliefs, especially those relating to esthetics, in different population groups. The evaluation of dental appearance and function in this study indicated that dental function is held in greater regard, and that the significance of dental appearance varies widely among dentists, dental technicians, and nondental subjects. PMID- 9728120 TI - Investigation of the dry and wet fatigue properties of three all-ceramic crown systems. AB - PURPOSE: The aim of this study was to investigate the influence of fatigue on the fracture strength of In-Ceram (Vita Zahnfabrik), Optimal Pressable Ceramic (Opc, Jeneric Pentron), and IPS Empress (Ivoclar-Vivadent) in both wet and dry environments. MATERIALS AND METHODS: Twenty-six crown shapes measuring 8.0 mm in diameter and 8.5 mm in height were fabricated for each ceramic system. For each ceramic system, 10 specimens were tested for fracture strength without fatiguing. A second group (8 specimens) was submitted to a fatigue and fracture test in dry conditions, and a third group (8 specimens) was fatigued and fractured in a wet environment using a mechanical testing machine (Instron). The results were statistically analyzed using a Mann-Whitney test. RESULTS: The results indicated that: (1) The facture strength for In-Ceram was significantly stronger than IPS Empress. No difference was found between In-Ceram and Opc, and Opc and IPS Empress. (2) The strength of the three ceramic systems decreased significantly after fatiguing in both dry and wet environments compared with the nonfatigued specimens. No difference was found between fatiguing in dry and wet environments. (3) For the three systems fatigued in a dry environment and then fracture tested, In-Ceram and Opc were significantly stronger than IPS Empress, but no difference was found between the three systems fatigued in a wet environment. CONCLUSION: Significant differences in the fracture strengths of the different systems investigated may be seen that result from both the nature of the system and the environment in which the specimens were fatigued. PMID- 9728121 TI - Occlusal appliance therapy in a short-term perspective in patients with temporomandibular disorders correlated to condyle position. AB - PURPOSE: The purposes of this study were to compare changes in the condyle-fossa relationship in patients with temporomandibular disorders of arthrogenous origin treated with either a stabilization or a control appliance in a double-blind controlled study, and to compare the changes in the condyle-fossa relationship with the short-term treatment effect in the two treatment groups. The radiographic appearance of the temporomandibular joint was also studied. MATERIALS AND METHODS: Fifty-eight patients with temporomandibular disorders of arthrogenous origin were assigned to two equally sized groups: a treatment group given a stabilization appliance; and a control group, given a control appliance. The study covered 10 weeks. The treatment outcome regarding changes in severity of temporomandibular joint pain on a verbal scale was compared to changes in the condyle-fossa relationship in horizontally corrected oblique lateral transcranial radiographs taken with and without the appliance. Condyle-fossa relationship and structural bone changes were observed before treatment in corrected lateral tomograms. RESULTS: The group treated with a stabilization appliance showed a changed condylar position significantly more often (P = 0.004) than the control group. Of the patients reporting a successful treatment outcome, significantly more (P = 0.006) showed a changed condyle position in the group treated with a stabilization appliance than in the group treated with a control appliance. CONCLUSION: In patients with temporomandibular disorders of arthrogenous origin, the short-term occlusal appliance therapy resulting in a changed condylar position gave relief of symptoms more often than if the condylar position was unchanged. PMID- 9728122 TI - Relationship between contact time measurements and PTV values when using the Periotest to measure implant stability. AB - PURPOSE: The Periotest is an electronic instrument that has been advocated for the measurement of implant stability and osseointegration. The aims of this investigation were to establish the relationship between contact time and PTV values when the Periotest was used to assess implants in vivo and in vitro, and to investigate the influence of the striking height of the Periotest handpiece and the length of implant abutment. MATERIALS AND METHODS: The accelerometer signal from a Periotest was captured and compared with the resulting PTV value. In vitro measurements of PTV and contract time were performed on a 3-mm abutment that had been attached to a 15-mm implant luted into an aluminium block, and were repeated on a patient in vivo. further measurements were made of the abutments of six implants in turn in the maxilla of the same patient. The standard abutment lengths on the implants were 3, 4 (x 2), 5.5 (x 2), and 7 mm. RESULTS: The results indicated that there was a linear relationship between contact time and PTV value for implants measured in vitro and in vivo. Greater scatter of the in vivo data was attributed to test and patient variables including striking position, distance, and damping as a result of the presence of soft tissue. There was a linear relationship between the PTV value and the striking height for implant measurements in vivo and in vitro. CONCLUSION: It can be concluded that the sensitivity of the Periotest to clinical variables including striking heights and handpiece angulation limit the application of the instrument as a clinical diagnostic aid to measure implant stability. PMID- 9728123 TI - Principles and applications of laser lithotripsy: experience with the holmium laser lithotrite. AB - INTRODUCTION AND OBJECTIVES: The initial clinical experience with holmium laser energy applied for endoscopic lithotripsy was positive. The current study is presented as a contrast to the preliminary findings and as a means of defining the clinical usefulness of this specific laser lithotrite. MATERIALS AND METHODS: Calculi were treated endoscopically with the holmium laser lithotriptor and data was gathered prospectively. The youngest patient in the series was a thirteen month--old who underwent percutaneous therapy, while the youngest patient on whom a retrograde endoscopic procedure was performed was a six-year old male patient with a proximal ureteral calculus. Lower water density, quartz fibers delivery systems were developed and employed. Fiber diameters ranged from 200-1000 micrograms. The smaller fibers were employed most commonly through the actively deflectable, flexible endoscope to facilitate treatment with maximum deflection. Larger fibers, with their much larger vaporization bubbles, were used through rigid endoscopes to debulk large stone burdens. RESULTS: A total of 210 patients with 249 calculi were treated. All major stone compositions were treated with minimal variation in laser efficiency. All but three of 109 ureteral calculi were treated in a retrograde fashion to completion (i.e., "stone free") in one sitting (97%). One-hundred thirteen renal stone burdens were treated with the holmium laser; 99 of these were treated solely in a retrograde fashion. Of the latter, 79 (80%) required only a single session. The combination of the actively deflectable, flexible ureteroscope and the 200-micrograms fiber facilitated treatment to completion of 38 to 45 lower-pole caliceal calculi (85%). The success of ureteropyeloscopic lithotripsy with the holmium laser for all intrarenal calculi, including staged or second sitting for large complex stone burdens, was 90%. Sixteen percutaneous procedures (13 renal and 3 ureteral calculi) employed the holmium laser as an endoscopic lithotrite. All 28 patients with large bladder calculi with a mean diameter of 41.8 mm were treated to completion in one sitting. Complications from holmium laser energy, including postoperative ureteral stricture disease, were not encountered in this series. CONCLUSIONS: Holmium laser energy is uniquely suited to treat urinary calculi safely regardless of stone size, location, or metabolic composition, and has particular efficacy in complex clinical presentations. PMID- 9728124 TI - Laser dosimetry studies in the prostate. AB - OBJECTIVE: To review the currently available data of photodynamic therapy (PDT) optical dosimetry for possible prostatic applications. SUMMARY BACKGROUND DATA: PDT is a new cancer treatment modality often used as an alternative tumor treatment method. Recently, PDT has been suggested as an alternative therapy for prostatic carcinoma and BPH. METHODS: PDT: utilizes light and a preadministered photosensitizer drug to achieve localized tumor control. This article reviews currently available data on optical dosimetry of PDT in both human and canine prostates. RESULTS: At 630 nm, a common wavelength used for Photofrin PDT, results indicate that light penetration is similar in cancerous and normal prostatic tissue. Because of limited light penetration, multiple fiber irradiation is necessary if eradicating the entire prostate glad is the ultimate goal. The available data also show that dynamic changes occur in light fluence rate distribution during PDT irradiation. CONCLUSIONS: PDT can be used to destroy prostatic tissue. Real-time optical dosimetry is necessary if accurate lesion volume control is desired. PMID- 9728125 TI - The holmium laser in urology. AB - OBJECTIVE: To review the physics related to the holmium laser, its laser-tissue interactions, and its application to the treatment of urological diseases. SUMMARY AND BACKGROUND DATA: The holmium: YAG laser is a solid-state, pulsed laser that emits light at 2100 nm. It combines the qualities of the carbon dioxide and neodymium:YAG lasers providing both tissue cutting and coagulation in a single device. Since the holmium wavelength can be transmitted down optical fibers, it is especially suited for endoscopic surgery. METHODS: The authors provide a review of the literature as it relates to the holmium laser and its application to urology. RESULTS: The holmium wavelength is strongly absorbed by water. Tissue ablation occurs superficially, providing for precise incision with a thermal injury zone ranging from 0.5 to 1.0 mm. This level of coagulation is sufficient for adequate hemostasis. The most common urologic applications of the holmium laser that have been reported include incision of urethral and ureteral strictures; ablation of superficial transitional cell carcinoma; bladder neck incision and prostate resection; and lithotripsy of urinary calculi. CONCLUSIONS: The holmium: YAG laser is a multi-purpose, multi-specialty surgical laser. It has been shown to be safe and effective for multiple soft tissue applications and stone fragmentation. Its utilization in urology is anticipated to increase with time as a result of these features. PMID- 9728126 TI - Application of the holmium:YAG laser for prostatectomy. AB - OBJECTIVE: The authors review the current knowledge regarding the application of the Holmium: YAG laser for prostatectomy. SUMMARY BACKGROUND DATA: Conventional surgical therapies for benign prostatic hyperplasia (BPH) are effective but associated with relatively high morbidity. Laser prostatectomy, using either Neodymium:YAG or potassium-titanyl-phosphate lasers, has emerged as a new and much safer operative approach to relieve symptoms of benign prostatic hyperplasia. However, these laser wavelengths possess key disadvantages that have limited their acceptability and dissemination in everyday urologic practice. METHODS: THE authors review their own extensive experience in the development of clinical application of Holmium: YAG laser technology for prostatectomy, as well as the published reports in the current medical literature now dealing with this subject. RESULTS: In multiple clinical trials, Holmium:YAG laser resection of the prostate has proven efficacious in relieving symptomatic BPH. Both objective urodynamic measures of voiding outcomes and symptomatic improvement have been shown to be equivalent to standard electrocautery resection of the prostate. At the same time, these studies have demonstrated the superior safety and hemostasis of Holmium:YAG laser prostatectomy compared to electrocautery resection, similar to prior laser prostatectomy procedure. Unlike prior forms of laser prostatectomy, Holmium:YAG laser resection of the prostate acutely removes all obstructing prostate tissue, so that the postoperative catheterization requirement is typically only overnight and improvement in voiding is immediate. Current operative techniques and the latest technological developments to facilitate Holmium:YAG laser prostatectomy are described. CONCLUSIONS: Holmium: YAG laser prostatectomy combines the best features of prior laser prostatectomy technologies, including minimal complications and morbidity, with the efficacy and immediacy of voiding outcomes associated with conventional electrocautery resection of the prostate. PMID- 9728127 TI - Minimally invasive therapies for the treatment of symptomatic benign prostatic hyperplasia: the University of Florida experience. AB - OBJECTIVE: To describe the University of Florida experience with minimally invasive therapies in the surgical treatment of benign prostatic hyperplasia (BPH). BACKGROUND DATA: Typically, the standard surgical treatment for symptomatic benign prostatic hyperplasia (BPH) has been transurethral resection of the prostate (TURP). Due to the morbidity associated with TURP, several minimally invasive therapies, such as laser, microwaves, high intensity focused ultrasound, and radiofrequency needle ablation, have been utilized to treat BPH. METHODS: The authors review their experience, along with that of others, with various forms of heat therapy in the treatment of BPH. RESULTS AND CONCLUSIONS: Although high intensity focused ultrasound (HIFU), interstitial laser, and microwaves procedures have been shown to be effective in the treatment of BPH, our experience has been with laser, VaporTrode, and TUNA. We found that VaporTrode and TUNA currently offer several advantages over many of the other modalities. PMID- 9728128 TI - Use of contact laser crystal tip firing Nd:YAG to relieve urinary outflow obstruction in male neurogenic bladder patients. AB - BACKGROUND AND OBJECTIVE: Endoscopic urologic procedures for transurethral prostatectomy (TURP), external sphincterotomy (TURS), bladder neck incision, and incising strictures using diathermy have resulted in excessive bleeding and risk of hyponatremia. This presentation is a review of a methodology developed to evaluate the use of contact laser crystal firing Nd:YAG laser. Details of the technique are presented. RESULTS: A review of 129 patients following laser TURS with 34% of these patients also needing TURP and 29% of patients also requiring TUIP has been done. Following contact laser endoscopic surgery, the catheter was removed in 24 hours. There was minimal to nil haemorrhage perioperatively and secondary haemorrhage was absent. CONCLUSIONS: The technique employing contact laser crystal provides an easy TURP, TURS, and stricture ablation. Follow up indicates durable results. PMID- 9728129 TI - Applications of the KTP laser in the treatment of posterior urethral valves, ureteroceles, and urethral strictures in the pediatric patient. AB - PURPOSE: We describes our experience using the potassium titanyl phosphate (KTP) 532 laser in treating posterior urethral valves, ureteroceles, and urethral strictures in the pediatric patient. METHODS: A retrospective chart review was performed from 1987 to 1997 on a total of 33 pediatric patients who underwent retrograde endoscopic treatment for posterior urethral valves (PUV), ureteroceles (UC), and urethral strictures using a KTP-532 laser. RESULTS: Overall, our success rate was excellent in the treatment of valves and ureteroceles. With a mean follow-up of three years in the PUV group, no urethral strictures of micturation abnormalities were seen. The majority of ureteroceles were decompressed and only half of our patients required and additional procedure. Our experience with urethral strictures, however, was not as promising. All of these patients ultimately required open urethral reconstruction. CONCLUSION: The desirable thermal characteristics of the KTP laser, along with minimal complications and the availability of delicate pediatric endoscopic instruments have made this operation optimally suited for treating posterior urethral valves and ureteroceles in infants. However, the advantages for treating urethral strictures in children with the laser still remains to be established. PMID- 9728130 TI - The application of laser techniques to vasectomy reversal surgery. AB - OBJECTIVE: A review of the application of laser technology to vasectomy reversal surgery. SUMMARY BACKGROUND: Modern methods of vasectomy reversal that employ microsurgical techniques have resulted in a high reported success rate. However, the procedure is tedious and time consuming. Laser technology offers the possibility of simplifying the procedure and reducing operative time, with possibly even better results. CONCLUSIONS: Application of lasers in vasovasostomy for vasectomy reversal is still in its early development. Several animal and human studies have been conducted with mixed results. Contained clinical trials will be necessary to prove the benefit of the laser in this surgical setting. PMID- 9728131 TI - Thermal lasers in urologic oncology. AB - Urology is a surgical specialty that relies heavily on the endoscopic approach for diagnosis and treatment of disease. Electrosurgical instruments have been the standard vehicle for endoscopic tumor ablation. Over the last 30 years a number of investigators have explored the use of the medical laser as either an alternative or an adjunct to standard electrosurgical techniques. The development of small caliber flexible and rigid endoscopic application. In addition, the potential for very limited and precise distribution of laser energy in targeted tissue is clinically appealing for endoscopic applications. In this article, we review the use of thermal laser in urologic oncology. PMID- 9728132 TI - Ureteroscopic laser treatment of upper urinary tract tumors. AB - OBJECTIVE: To summarize the present status of ureteroscopic laser treatment of upper urinary tract tumors. SUMMARY AND BACKGROUND DATA: Small diameter rigid and flexible ureteroscopes can provide convenient access to the upper urinary tract. The small diameter of the instruments and the working channels are ideally suited for the placement of laser fibers as an intraluminal ablative technique. METHODS: The authors reviewed the literature and their own experience with laser treatment of upper tract tumors for the description of instruments, techniques, and the results achieved. RESULTS: At least 12 reports have presented the results after using lasers for treating upper tract tumors. The Nd:YAG laser has been used in most series but more recently, the holmium:YAG laser has become available. Each laser has particular advantages and each can be delivered along the same low water content quartz fibers. Less scarring with stricture formation has been reported after use of the laser versus electrofulguration. Among the series reported, local recurrences occurred in 33% of patients with renal pelvic tumors or ureteral tumors. In the authors' experience, the holmium:YAG laser was most frequently used and the combination of holmium and Nd:YAG was nearly as common. The Nd:YAG was used alone in less than 10% of patients. CONCLUSIONS: The ureteroscopic treatment of upper tract transitional cell carcinoma is a reasonable alternative to surgical removal in many patients. It provides a major advantage in patients with specific indications for conservative therapy and may be a reasonable elective therapy in others with small, low grade, tumors. The holmium laser alone or in combination with the Nd:YAG laser has become our primary mode of therapy. PMID- 9728133 TI - Photodynamic therapy (PDT) in the treatment of patients with resistant superficial bladder cancer: a long-term experience. AB - INTRODUCTION AND OBJECTIVE: Photodynamic therapy (PDT) combines a photosensitizer such as Photofrin with red laser light (630 nm) to destroy cancer cells. Investigators have reported effectiveness of PDT in the management of patients with recurrent superficial bladder cancer. We retrospectively reviewed our experience in 58 patients to assess the long-term role of PDT in the management of resistant superficial transitional cell carcinoma (TCC) including Ta, T1, and refractory carcinoma in situ (CIS) of the urinary bladder. MATERIALS AND METHODS: All 58 patients had failed at least one course of standard intravesical therapy or had contraindication for intravesical chemo- or immunotherapy. Patients with malignancy present (Ta-T1/Grade I-III, CIS) were accepted for ablative PDT. Patients undergoing prophylactic PDT after complete resection were confirmed to be tumor-free by cystoscopy and bladder was cytology before PDT. Post-PDT evaluations included weekly telephone contact to assess acute adverse reactions and assessment of efficacy and bladder toxicity at three months and quarterly thereafter. RESULTS: These 58 patients underwent a single PDT treatment with 2.0 or 1.5 mg/kg of Photofrin and 10-60 J/cm2 light (630 nm). At three months, complete response rates were 84% and 75% for residual resistant papillary TCC and refractory CIS respectively; and 90% of patients treated prophylactically had not had recurrences. At a median followup of 50 months (range 9-110), 59% (34/58) of the responders are alive, with 31/34 still disease-free. CONCLUSION: PDT using 1.5 mg/kg of Photofrin and 15 J/cm2 of light (630 nm) should be considered a safe and effective treatment for refractory CIS or recurrent papillary TCC. PMID- 9728135 TI - Conflict of interest: a dilemma that requires clarification. PMID- 9728134 TI - Aesthetic restoration with full-coverage porcelain veneers and a Ceromer/fiber reinforced composite framework: a case report. AB - Due to the recent evolution of dental materials and procedures, the restoration of a particular clinical condition can be accomplished by various alternatives. While the use of integrated treatment modalities permit the restoration team to utilize the benefits of each material simultaneously, it requires the development of a comprehensive preoperative treatment plan to address the specific concerns of each region or material utilized. This article describes the use of a multidisciplinary approach to restore proper occlusion, function, and aesthetics in a patient with worn dentition. PMID- 9728136 TI - Use of bonded porcelain restorations for nonorthodontic realignment of the anterior maxilla. PMID- 9728137 TI - Clinical and laboratory considerations in the use of a new all-ceramic restorative system. AB - Ceramic systems are continually under development in an effort to refine their clinical application. An all-ceramic full-coverage crown system (Procera, Nobel Biocare, Westmont, IL) that utilizes computer technology and industrial presses to fabricate precise copings has recently been introduced. Using this system, aluminous porcelain is baked over a high-strength, high-purity aluminum oxide coping to fabricate a functional, biocompatible, and aesthetic restoration. This article presents a discussion of the clinical and laboratory considerations that are involved in the utilization of this all-ceramic system. PMID- 9728138 TI - The skeleton buildup technique: a systematic approach to the three-dimensional control of shade and shape. AB - Due to the limitations of current shade-matching systems, shade communication has proven inadequate. Techniques for the fabrication of porcelain crowns that match the natural definition must address numerous factors. While all-ceramic restorations are indicated for the rehabilitation of the anterior dentition, these modalities are problematic in the restoration of teeth with discolored substrates. This article presents a systematic procedure for the three dimensional fabrication of porcelain restorations. Techniques for building porcelain and altering the translucency of ceramic cores are also presented. PMID- 9728139 TI - Utilization of leucite glass ceramic veneers for complex case rehabilitation. PMID- 9728140 TI - Patients' insecurities. PMID- 9728141 TI - Pulpal regeneration. PMID- 9728142 TI - Full-arch rehabilitation utilizing pressed ceramic veneers and full-coverage crowns. AB - A variety of all-ceramic systems has been developed to address the limitations exhibited by conventional porcelain-fused-to-metal restorations. The fabrication principles used for pressed ceramic materials requires the application of a vacuum pressure, which results in a restoration with enhanced optical and physical properties. Due to the strength and versatility of these restorations, the indications for the use of pressed all-ceramic restorations continues to expand. This article describes the application of pressed ceramic restorations for comprehensive rehabilitation of the maxillary arch. PMID- 9728144 TI - The practice of dentistry: to have or to be. PMID- 9728143 TI - Periodontal implications in implant treatment planning for aesthetic results. AB - Insufficient hard and soft tissue height and width can be a repercussion of tooth loss or a result of postoperative healing following implant surgery. Insufficient bone can preclude proper implant positioning, while inadequately treated soft tissue will not exhibit a gingival appearance similar to that of the adjacent teeth. If hard and soft tissue discrepancies are not corrected by regenerative techniques, the replaced tooth appears long or overbulked gingivally. In order to create hard and soft tissue harmony, an understanding of the biological variables and periodontal implications is necessary. PMID- 9728145 TI - Treatment planning extensively broken-down mandibular molars for post-and-core fabrication. AB - Because of their unique radicular anatomy, mandibular molars require careful treatment planning for post-and-core fabrication. Indications for post-and-core fabrication for endodontically treated mandibular molars are discussed, and immediate and cast post-and-core techniques are reviewed. An alternative design for a cast post and core is also presented. PMID- 9728146 TI - Microleakage of tooth-colored restorations with a beveled gingival margin. AB - OBJECTIVE: Microleakage of tooth-colored restorative systems was tested in preparations with and without beveled gingival margins. METHOD AND MATERIALS: A resin composite, Z-100, and two "compomer" restoratives, Dyract and Geristore, with their accompanying adhesive systems, were placed in nonretentive cervical cavity preparations (at the cementoenamel junction), with and without a beveled gingival margin (dentin or cementum) and beveled occlusal (enamel) margins in extracted bovine teeth. Microleakage was assessed as the ratio of the extent of methylene blue dye penetration at the tooth-restoration interface of the length of the wall. RESULTS: Z-100 restorations, without a gingival bevel, exhibited significantly less microleakage along the gingival wall and less microleakage overall than did the other materials. Dyract restorations without a gingival bevel and placed without a surface conditioner displayed greater microleakage overall than did the other groups. CONCLUSION: With all materials, Class V restorations with gingival bevels displayed greater microleakage than did nonbeveled margins. PMID- 9728147 TI - Inhibition of demineralization in vitro around amalgam restorations. AB - OBJECTIVE: The purpose of this in vitro study was to evaluate some forms of preventing or avoiding demineralization within enamel cavity walls adjacent to amalgam restorations. METHODS AND MATERIALS: Third molar teeth were sectioned to obtain 72 specimens, divided into one control and five experimental groups: amalgam only; varnish plus amalgam; acidulated phosphate fluoride plus amalgam; adhesive amalgam; glass-ionomer cement plus amalgam; control (amalgam only, not subjected to a demineralization challenge). The experimental groups were subjected to PH and thermal cycling and then submitted to enamel hardness determinations. RESULTS: Significant differences between the treatment groups revealed that the bonded amalgam technique offered the best resistance to demineralization. The use of cavity varnish resulted in greater mineral loss than amalgam placed alone. CONCLUSION: The use of an adhesive system, glass-ionomer cement, or acidulated phosphate fluoride under amalgam restorations may interfere with development of secondary caries. PMID- 9728148 TI - Development and clinical application of titanium minipins for fixation of nonresorbable barrier membranes. AB - Clinical applications of the principle of guided bone regeneration in oral and maxillofacial surgery include preimplant reconstruction of atrophied bone defects and coverage of endosseous implants that were incompletely covered by bone through primary intention. Nonresorbable polytetrafluoroethylene membranes are used. Over the past several years, the use of periosteal sutures to fix membranes has been supplemental or replaced by the use of metallic fixation systems. Five year clinical results with a membrane pin set developed for fixation of such membranes are reported. Application of the titanium pin limits the relative movement between the membrane and surrounding bone and/or between and surrounding soft tissue flaps. Moreover, the titanium pin expands the range of applications for such membranes, particularly to topographically complicated bone defects at sites where clinically secure and biologically functioning placement of the membrane is not always easy. PMID- 9728149 TI - Efficacy of a synthetic polymer saliva substitute in reducing oral complaints of patients suffering from irradiation-induced xerostomia. AB - OBJECTIVE: A saliva substitute based on polyglycerylmethacrylate, lactoperoxidase, and glucose oxidase (Oral Balance) has been developed. The aim of this study was to evaluate the effect of Oral Balance on the dryness-related oral complaints in patients suffering from irradiation-induced xerostomia. METHOD AND MATERIALS: The efficacy of Oral Balance on the dryness-related complaints of 28 patients was assessed by means of self-administered questionnaires. Each patient completed an initial questionnaire about dryness-related symptoms and then was given the moistening gel. The patients were instructed to apply the gel as often as desired. After 2 weeks and 3 months, the patients were asked to complete a progress questionnaire. The severity of xerostomia was measured with a saliva absorption method. RESULTS: All patients suffered from moderate-to-severe xerostomia, the severity of which did not change during the experimental period. Three patients did not complete the study. In the other 25 patients, the application of Oral Balance tend to diminish the sensation of oral dryness and improve oral functioning. Statistically significant reduction of the dryness related complaints was observed only in the patients suffering from severe xerostomia. CONCLUSION: Use of Oral Balance is of potential benefit in patients suffering from severe xerostomia. PMID- 9728150 TI - The importance of occlusal balance in the control of complete dentures. AB - OBJECTIVE: The importance of occlusal balance to the control of complete dentures during function was assessed. METHODS AND MATERIALS: The complete dentures of five patients who were having difficulty controlling their prostheses were accurately duplicated. The artificial teeth were replaced with occlusally balanced teeth. No other changes were made. Patients were asked to report their experiences with the new dentures after 1 week, 3 weeks, and 6 weeks. RESULTS: By the end of 6 weeks, improvement in denture stability and eating comfort were reported by all patients. CONCLUSION: Improvements occurred when the occlusion was balanced, despite existing jaw relationship errors, fitting inaccuracies, and peripheral extension errors. PMID- 9728151 TI - Transmigration of mandibular canines: report of six cases and review of the literature. AB - Six young adults were found to have transmigrated mandibular canines. One patient presented with two transmigrated canines. Of the seven impacted teeth, the left mandibular canines was involved in five instances and the right two. In all patients, the primary canine was present in the dental arch. A supernumerary tooth was disclosed on the panoramic radiograph of two patients. Five patients underwent surgical removal of the transmigrated tooth from an intraoral approach. One patient experienced transient postoperative paresthesia in the zone innervated by the mental nerve. When the transmigrated canine is accessible, and especially if it is symptomatic, removal of unerupted tooth is recommended. Otherwise, it should be left alone and kept under observation. PMID- 9728152 TI - Improving dentist-patient relations. PMID- 9728154 TI - On the present position of treatment of fractures (with special plates). 1912. PMID- 9728155 TI - Acromioclavicular dislocation. Conservative or surgical therapy. AB - A literature review was performed to clarify available information which influences decisions whether to advise a young adult patient to undergo surgery for a severely displaced acromioclavicular dislocation. Twenty-four papers were retrieved yielding 1172 patients of whom the mean followup for the 833 surgically treated patients was 43.7 months and not surgically treated was 60.4 months. Of the 24 papers, only five reported surgical and conservative outcomes; two of these papers used prospective randomized methodology and three used nonrandomized methodology. Fourteen papers reported surgical outcome only and five papers reported conservative outcome only. Overall, 88% of surgically treated patients and 87% of nonsurgically treated patients had a satisfactory outcome. Complications most commonly listed were (surgically treated versus nonsurgically treated): need for further surgery (59% versus 6%), infection (6% versus 1%), and deformity (3% versus 37%). Return to activity was no quicker with surgery. Pain was not any more common without surgery. Range of movement was more frequently normal or near normal without surgery (95% versus 86% if surgically treated) and so was strength (92% versus 87%). Meta-analysis of the four studies including data from surgical and conservative therapy showed on significant benefit from surgery. Power studies suggest that to show a statistically significant benefit from surgery, large studies would be required, which, given the relative incidence of these injuries, would probably be multicenter and therefore vulnerable to methodologic difficulties. There does not seem to be any reason to recommend an operative procedure to a patient with a Rockwood et al Type III injury based on the evidence currently available. PMID- 9728156 TI - Assessment and management of three-and four-part proximal humeral fractures. AB - Three-and four-part comminuted fractures of the proximal humerus are difficult and technically demanding to treat. The various treatment methods reported in the literature are reviewed. It is recommended that three-part fractures be treated with open reduction and internal fixation. Four-part fractures in the younger, active patient also can be treated successfully with open reduction and internal fixation. However, in the elderly and in the patient with osteoporosis, a hemiarthroplasty is the treatment of choice. There is a need for universal agreement on a scoring system for measuring outcome in these fractures to allow a meaningful comparison between reported treatment methods. PMID- 9728157 TI - Controversies regarding radial neck fractures in children. AB - Fractures of the radial neck in children are not uncommon, yet several aspects of their management remain controversial. Until a consensus is reached regarding the determination of displacement, acceptability of initial angulation, treatment, and outcome, the complication rate of these fractures will remain high. The authors recommend measuring the displacement as the angle between a line perpendicular to the articular surface of the radial head with a line down the shaft of the proximal radius. Fractures that are angulated less than 30 degrees require immobilization alone. Fractures angulated more than 30 degrees should be treated with an attempt at closed reduction. If closed reduction fails, a percutaneous reduction should be attempted before open reduction. Internal fixation should be performed using an oblique extraarticular Kirshner wire for all unstable fractures. Grading outcome based on range of motion and the presence or absence of pain is recommended. It is hoped that once the controversies surrounding these fractures are resolved, the long term results of these troublesome injuries will improve. PMID- 9728158 TI - Comminuted fractures of the radial head. Arthroplasty versus internal fixation. AB - Fractures of the radial head continue to challenge orthopaedic surgeons. Fortunately, most simple uncomplicated fractures treated non-operatively with emphasis on early motion achieve good results. Treatment of more complex fractures remains controversial, however. When simple radial head excision is contraindicated, choosing between open reduction and internal fixation and radial head replacement remains difficult. A review of the literature does not provide definite guidelines, but suggest that fracture complexity and technique are critical for success. This paper is not intended to review the treatment of radial head fractures, but rather to focus on choosing between replacement versus internal fixation when preservation of radial head mechanics is indicated. PMID- 9728159 TI - Management of comminuted fractures of the distal radius in the adult. Conservative or surgical? AB - Displaced fractures of the distal radius are difficult to treat successfully by traditional nonoperative methods. The goal in the management of these fractures is to achieve extraarticular alignment and an articular step off of less than 2 mm. Cast immobilization has been supplemented with pins and plaster technique and external fixators. Percutaneous are limited open reduction techniques, combined with wrist arthroscopy, have been shown to be useful in the management of intraarticular distal radius fractures. Despite these advances, there are still a significant number of fractures in which the articular surface cannot be reconstructed without open reduction and internal fixation. The main objective is to restore articular integrity as perfectly as possible. Attention to meticulous surgical technique will facilitate good results. When articular restoration cannot be accomplished, early arthrodesis or arthroplasty should be indicated. In the absence of osteoarthritis, intraarticular osteotomy can be used for intraarticular malunions with a step off greater than 2 mm. Radius malalignment usually requires a dorsal opening wedge osteotomy, insertion of a corticocancellous graft, and a dorsal buttress plate. Early recognition and treatment of distal radioulnar joint injuries associated with fractures of the distal radius are paramount to reduce the incidence of painful sequelae and functional deficits. PMID- 9728160 TI - Controversies in the treatment of fingertip amputations. Conservative versus surgical reconstruction. AB - Fingertip amputations are the most common type of amputation injury in the upper extremity. These injuries are either seen in the emergency room or in an office setting. These lesions are very frequent and require precise wound care for optimal results. The longer finger and the thumb, being the most distal and independent mobile parts of the hand are affected very often by these kind of injuries. Treatment of fingertip injuries is a continuous focus of controversy among hand and orthopaedic surgeons. Different treatment options have been described, depending on the affected segment and finger, type of lesion, gender and age of the patient, location, size, and depth of the defect. The indications, advantages, and disadvantages of several reconstructive procedures for fingertip injuries have been described. The techniques themselves are not described in detail. PMID- 9728161 TI - Controversies in acetabular fractures. AB - Open reduction and internal fixation has become the standard of care for the treatment of most displaced acetabular fractures. As surgical techniques have become refined, long term results of surgical fixation have improved. During the past 10 to 15 years, several controversies have surfaced in the orthopaedic literature regarding the treatment of acetabular fractures. The recent literature regarding acetabular fixation was reviewed. Controversies include the most efficacious surgical approach for complex acetabular fractures; the effectiveness of intraoperative sciatic nerve monitoring; the most effective method of prophylaxis against deep vein thrombosis; and the indications for and method of prophylaxis against heterotopic bone formation. PMID- 9728162 TI - Unipolar or bipolar prosthesis for the displaced intracapsular hip fracture? An unanswered question. AB - Modular bipolar prostheses were developed to address the problems of loosening, cartilage wear, and protrusio which were seen with single unit endoprostheses. Modular unipolar prostheses address many of these problems and are significantly less expensive than the bipolar prosthesis. Recent data suggest that use of the modular unipolar prosthesis is indicated in elderly patients with low demands. PMID- 9728163 TI - Displaced fractures of the femoral shaft in children. Unique features and therapeutic options. AB - The decision analysis for managing femoral shaft fractures in children should included such factors as the possibility of child abuse, overgrowth, and the potential for remodeling. Direct and indirect costs must be understood. Factors to consider in determining treatment include the age of the child, the extent of the soft tissue injury, and associated injuries. Non-operative methods, universally used in the past to treat these injuries, still are indicated, but operative modalities should be considered for a greater number of pediatric femoral fractures. Early enthusiasm for external fixation and rigid intramedullary rodding has been tempered by a greater awareness of their particular complications. The role of flexible intramedullary rodding, however, is expanding. PMID- 9728164 TI - Treatment of complex lateral plateau fractures using Ilizarov techniques. AB - Fourteen high energy atypical Schatzker Types I and II fractures were treated using a combination of contemporary internal fixation techniques and Ilizarov methodologies. Fracture were atypical if the primary lateral condylar fracture line extended anteromedially with complete detachment of the tibial tubercle, or posteromedially with extension into the medial or posterior medial tibial condyle, both in association with comminution of the lateral condyle and impaction of the lateral articular surface. Preoperative traction computed tomography scans were used to direct incisions to the area of joint involvement. Fixation included hook plate stabilization of anterior tubercle fragments in five fractures. Adjunctive lateral, medial, or posterior antiglide plates were used where olive wire stabilization was contraindicated because of anatomic constraints. Fractures were neutralized by a three-ring Ilizarov external fixator. At followup, average 19.2 months (range, 8-67 months), all fractures had healed, and 85% had good or excellent knee scores (Knee Society clinical rating scale). Five patients had minor pin tract complications and one patient had a superficial wound slough. This combined approach has shown excellent results for this complex fracture pattern without the severe soft tissue complications associated with internal fixation techniques for high energy fractures. PMID- 9728165 TI - Management of calcaneal fractures in adults. Conservative versus operative treatment. AB - Significant progress had been made in the management of calcaneal fractures. This is reflected in the marked decrease in complication rates associated with the current intervention of these potentially devastating injuries. The treatment priorities that are key to achieve best results in a displaced calcaneal fracture are an anatomic reconstruction of the entire calcaneus including articular surfaces, height, alignment, and length, with a function directed postoperative management. The value of these priorities is confirmed by long term followup results. Conservative treatment should be considered only in cases of extraarticular fractures, in cases of minor displaced intraarticular fractures in patients who are nonambulatory, and in cases where there is a clear contraindication for surgery. An anatomic reconstruction of an os calcis fracture is difficult to obtain. In two-part fractures, according to the classification described by Sanders et al, an anatomic reduction is obtainable in more than 80% of cases. However, if the articular cartilage damage that is typically present is considered, a 70% rate of good to excellent clinical results is more realistic. In three-part fractures, anatomic reduction is attainable in approximately 60% of cases with a 70% rate of good results. These two subgroups comprise approximately 90% of all calcaneal fractures. It has been put into practice recently to optimize the extended lateral approach using posteromedial and anterolateral windows, so that an anatomic reduction can be achieved in more than 60% of os calcis fractures considered as Type III according to the classification described by Sanders et al. Additional scientific work in this area of trauma orthopaedics would benefit most from a general consensus on a fracture classification system and on a clinical scoring system, with 5-year followup studies using these treatment methods and evaluation systems. PMID- 9728166 TI - Surgical management of acute tarsometatarsal fracture dislocation in the adult. AB - Injuries to the tarsometatarsal joint involving fracture dislocations are uncommon and are often referred to as Lisfranc lesions after the French field surgeon in the Napoleonic Wars. Despite the infrequency of this serious injury, they have the potential for chronic disability and require prompt, accurate diagnosis and precise anatomic reduction to minimize long term disability. A review of the literature shows that opinions differ as to the most appropriate method of treatment for these injuries, be it closed or open reduction, but most authors agree that it is imperative to achieve precise anatomic reduction. PMID- 9728167 TI - Posteroinferior acromioclavicular dislocation with supraspinatus tear. A case report. AB - A 20-year-old man was treated for posteroinferior acromioclavicular dislocation. The diagnosis was based on standard radiographs and intraoperative findings. The distal end of the clavicle had impaled the supraspinatus muscle. Open reduction was performed 2 weeks after injury, followed by wire fixation of the acromioclavicular joint and repair of the torn superior acromioclavicular ligament and coracoclavicular ligaments. Two years after the procedure, standard radiographs revealed normal anatomic alignment of the acromioclavicular joint, with pain free range of motion. Active elevation in the scapular plane was 180 degrees, active external rotation was 80 degrees in the anatomic position, and passive internal rotation was to the T5 vertebra. The patient returned to playing baseball and tennis and was satisfied with the postoperative result. PMID- 9728168 TI - Hemodynamic measurement in the femoral head using laser Doppler. AB - The author used a laser Doppler device to measure hemodynamics in the femoral head. Twenty-eight patients with femoral neck fractures, 16 patients with intertrochanteric fractures, and 14 patients with osteoarthritis were studied. Through a lateral approach, a small hole was drilled into the femoral head using a burr, and a probe was inserted to measure intramedullary flow. The flow measurements were high and sinusoidal in shape (corresponding with the heart rate) in all Garden 1, all Garden 2, and in four of six Garden 3 femoral neck fractures. Flow measurements were low and not sinusoidal in two of six Gardens 3 and all Garden 4 fractures. Hemodynamic values were high in all 16 patients with intertrochanteric fractures and all 14 patients with osteoarthritis. Although the posterior column arteries were cut while the intramedullary hemodynamics were being measured in patients with osteoarthritis of the hips, the measured values still were high. The postero-lateral area of the femoral head was fed compensatorily by the inferior retinacular arteries. The laser Doppler is useful in estimating circulatory compromise. PMID- 9728169 TI - Hemarthrosis and hip joint pressure in femoral neck fractures. AB - In a prospective clinical study the intraarticular pressure of 55 patients with intracapsular femoral neck fractures was measured intraoperatively with the hip in different positions. Intraarticular hemarthrosis was quantified by a preoperative sonography examination. In 75% of the patients, increased intraarticular pressure caused by the hemarthrosis was found. The spontaneous median pressure increased significantly from 22 mm Hg with extension (28 mm Hg) and internal rotation of the hip joint (56 mm Hg). The lowest pressure was found in 70 degrees flexion (15 mm Hg). The median pressures increased within the first 24 hours after injury from 26 mm Hg in the first 6 hours to 46 mm Hg from 7 to 24 hours. Even in the first and second weeks after trauma, increased median pressures were detected (8.5 mm Hg and 13 mm Hg, respectively). No significant difference was found between undisplaced and displaced fracture types. Because increased joint pressure in other studies correlates with reduced perfusion of the femoral head, it can be deduced that reduction maneuvers without capsulotomy can compromise the circulation of the femoral head. Capsulotomy and osteosynthesis of the femoral neck at the earliest time possible is the best prophylaxis of tamponade. If the osteosynthesis is delayed, a preoperative sonography after admission and a control sonogram after 6 hours is recommended. In the event of relevant hemarthrosis, immediate therapeutic drainage is suggested for patients who will receive joint conserving osteosynthesis. PMID- 9728170 TI - Operative hip arthroscopy. AB - Twenty patients underwent operative arthroscopic procedures of the hip joint. All procedures were performed with the patient in the supine position on a standard fracture table using fluoroscopy through three arthroscopic portals (anterolateral, anterior paratrochanteric, and posterior paratrochanteric). The initial indications were therapeutic in 16 patients: loose bodies in four, synovial chondromatosis in three, rheumatoid arthritis in five, ankylosing spondylitis in one, septic arthritis in one, avascular necrosis of femoral head in one, and primary osteoarthritis in one. In four patients who had unexplained hip pain, the initial indications were diagnostic: minimal synovial change was seen in two patients, a synovial chondromatosis was present in another, and a tear of the acetabular labrum and hypertrophy of ligament teres were present in a fourth patient. In one patient who had primary osteoarthritis, the insertion of the arthroscopic instrument into the hip joint failed because of profuse osteophytes along the acetabular rim. Twelve of the 19 patients showed significant improvement of the symptoms after the arthroscopic procedure, but seven patients had no benefit from the procedure. One patient had a postoperative reflex sympathetic dystrophy. PMID- 9728171 TI - Total hip replacement rates are higher among Caucasians than Asians in Hawaii. AB - The etiology of hip osteoarthritis remains unknown but may involve genetic or lifestyle factors. Most cases of total hip replacement are performed because of osteoarthritis. To examine possible ethnic differences, hospital records in Hawaii from 1985 to 1989 were reviewed. Preoperative radiographs were reviewed for a subset of patients to ascertain the reason for total hip replacement. Osteoarthritis accounted for a greater percentage of total hip replacements among whites (59% for women and 66% for men) than among Japanese (36% of women and 30% of men). The incidence of total hip replacement for whites was three to 25 times greater than that of other ethnic groups (Japanese, Chinese, Filipino, and Hawaiians). For example, the risk of total hip replacement for white women 40 years to 84 years of age was 4.4%, compared with 1.1% for Japanese women and 1.7% for Chinese women of the same age group. Compared with published data, the incidence was similar for Chinese in Hawaii and San Francisco; however; whites in Hawaii had a total hip replacement incidence less than half that of whites in San Francisco. Lifestyle differences might account for the lower incidence of total hip replacement for whites in Hawaii, compared with those in San Francisco. The lower incidence among Asians suggests a possible genetic basis for osteoarthritis. PMID- 9728172 TI - Prediction of postoperative knee flexion in Insall-Burstein II total knee arthroplasty. AB - Postoperative knee flexion in patients undergoing Insall-Burstein-II total knee arthroplasty at 2 years was evaluated regarding two basic questions: what groups of patients gain or lose the most flexion and what groups of patients have the best or worst postoperative flexion. Thirteen preoperative variables (maximum flexion, flexion arc, tibiofemoral angle, quadriceps strength, extensor lag, Knee Society score, Knee Society patient assessment, gender, age, height, weight, diagnosis, and surgeon) and four postoperative variable (leg length change, tibiofemoral angle, distance from patella to the joint line, and the tibial prosthesis anteroposterior translation on a lateral radiograph) were used in an attempt to explain postoperative flexion. The analysis was performed on 164 consecutive Insall-Burstein-II total knees in which the data were gathered prospectively on a time oriented medical record database. A regression tree analysis was used to identify several groups of patients, characterized by preoperative factor values, who had markedly above average performance on postoperative flexion. The preoperative factors identified include preoperative flexion, flexion arc, tibiofemoral angle, extensor lag, diagnosis, and age. The only postoperative variable of significance was tibiofemoral angle. Among the potential determinants of postoperative flexion that failed to appear predictive were the Knee Society scores and surgeon. Preoperative flexion is known to be a critical determinant of postoperative flexion in total knee replacement. However, in the current study, preoperative flexion accounted for only half of the difference between the best (122 degrees) and the worst (88 degrees) group, as determined with regression tree analysis. PMID- 9728173 TI - Tibial osteotomy for varus gonarthrosis. A 10- to 21-year followup study. AB - From 1975 to 1986, 102 high tibial osteotomies for varus gonarthrosis were performed in 99 patients. Fifty-eight patients (60 knees) were reviewed at an average followup of 15 years (range, 10-21 years); of the remaining 41 patients, seven had a knee replacement, 18 had died, and 16 were lost to followup. The results, assessed according to the scoring system of the Hospital for Special Surgery and including the seven patients who underwent a knee replacement, were excellent or good in 37 (55%) knees and fair or poor in 30 (45%). Twenty-six patients of the current study previously were reviewed in 1986, with an average followup of 8 years, using the same clinical and radiographic criteria. In this group of 26 patients, excellent and good results decreased from 73% in 1986 to 46% in 1996. The knees in these 26 patients with a followup greater than 15 years had a statistically significant higher percentage of fair and poor results. No statistically significant differences in the results were found according to the amount of correction. Radiographic controls at followup were available for 45 of the 60 knees; a loss of correction greater than 5 degrees was observed in 11 knees. The results of this long-term followup study show that high tibial osteotomy for gonarthrosis allows a long period (range, 10-15 years) of relief of pain, good range of motion, and function in a large number of patients. Results tend to deteriorate with time, particularly after 15 years. PMID- 9728174 TI - Effect of partial release of the posterior cruciate ligament in total knee arthroplasty. AB - Appropriate tension of the posterior cruciate ligament, which often is tight in deep flexion, is difficult to achieve after posterior cruciate ligament retaining total knee arthroplasty. Kinematics and maximum flexion after partial release of the posterior cruciate ligament were evaluated in this study. A partial release improved the maximum flexion angle and maintained anteroposterior stability without causing undesirable changes in kinematics, whereas full resection of the posterior cruciate ligament caused unfavorable anteroposterior instability. Partial posterior cruciate ligament release eliminated excessive rollback movement caused by a tight posterior cruciate ligament and also shifted the point of articular surface contact anteriorly. These results indicate that partial release of the posterior cruciate ligament may improve knee function in patients with a tight posterior cruciate ligament after total knee arthroplasty. PMID- 9728175 TI - Computed tomography for femoral and tibial torsion in children with clubfoot. AB - Forty-seven children with 70 clubfeet had computed tomography studies performed to determine the degree of femoral, tibial, and total limb torsion in both lower limbs. The total limb torsion angle (angle between the axis of the femoral neck and the axis of the ankle), which describes the relationship between femoral and tibial torsion, was used to evaluate the whole rotational deformity of the lower limb. The children were between the ages of 2 and 10 years (mean, 5 years) at the time of the computed tomography study. The mean femoral torsion was 25 degrees in the limbs with a clubfoot and 23 degrees in the contralateral limbs of patients with a unilateral clubfoot. The mean tibial torsion was 25 degrees in the limbs with a clubfoot and 24 degrees in the contralateral limb of patients with a unilateral clubfoot. The authors observed decreases of anterior femoral torsion corresponding to increases in age, consistent with the observations made by other authors of studies of children without clubfoot. External tibial torsion increased with age, with similar values in limbs with and without clubfoot. Ten limbs (nine with clubfoot, one without clubfoot) had femoral torsion greater than the means plus one standard deviation and 12 limbs (eight with clubfoot, four without clubfoot) had tibial torsion less than the means minus one standard deviation. The authors found four limbs (all with clubfoot) in three patients with lower than the mean minus one standard deviation of the total limb torsion angle (intoeing). Overall, there was no appreciable difference in the amount of femoral or tibial torsion in limbs with and without a clubfoot. PMID- 9728176 TI - Head and neck replacement endoprosthesis for pathologic proximal femoral lesions. AB - Records of 28 patients with pathologic lesions in the proximal femur treated by implantation of a femoral head and neck replacement prosthesis between 1984 and 1995 were reviewed. Mean clinical followup was 47.8 months in the eight living patients and 15.8 months in the 20 patients who had died. The underlying diagnosis was metastatic disease or myeloma in 22 patients. The most frequently occurring indication for implantation of this device was a pathologic fracture in 26 patients (18 displaced, eight impending), followed by resection and reconstruction in two patients. All femoral components were cemented: 23 were bipolar hemiarthroplasties and five were total hip arthroplasties. Implant survivorship was good (93%), with only two prostheses removed during the followup period, both for infection. However, radiographic analysis revealed increasing lucencies with time, particularly in the most proximal zones, resulting in radiographic failure in an additional case. Deep infection occurred in three cases, leading to resection arthroplasty in two patients. Periprosthetic fractures occurred in three cases, but only one occurred intraoperatively. Despite a high complication rate, the good implant survival during the shortened life span of these patients supports the continued use of femoral head and neck replacement prostheses in this population. PMID- 9728177 TI - Giant cell tumor of lumbar vertebra. A case report with 13-year followup. AB - A combined one stage anterior and posterior approach was used to excise a giant cell tumor involving the second lumbar vertebra. The tumor involved the anterior and posterior elements of the vertebra. A wide excision was feasible with immediate stabilization using the Luque instrumentation posteriorly and fibular strut grafts supporting the vertebral bodies anteriorly. The followup period was 13 years. There is no recurrence, and the patient has no symptoms of disease, and the fibular grafts are fully incorporated. PMID- 9728178 TI - Treatment of nonunion of the humerus using the Ilizarov external fixator. AB - Ilizarov's method of monofocal compression was used in 30 humeri with a diaphyseal pseudarthrosis. Twenty-one patients had previous surgery but had loosening of the osteosynthesis material. Nine patients initially were treated with a hanging cast, resulting in interfragmentary distraction. Fourteen nonunions were hypertrophic, and 16 were atrophic, of which six were infected. A complete circular frame was used only in the first nine patients, whereas the remaining 21 patients were treated with the modified semicircular fixator. Union was obtained in all but two patients, with an average consolidation time of 4.5 months (range, 2.5-10 months). No patient required additional bone grafting. Apart from superficial pin tract infection seen in most of the patients, three had a minor temporary sensory neurologic problem. Four patients experienced a second fracture after removal of the fixator that required a second application of an Ilizarov frame. Although similar results with regard to union are reported after plate osteosynthesis, there was no radial nerve palsy or deep infection in this series, indicating that the treatment by the Ilizarov technique is associated with less complications. The authors' findings suggest that the Ilizarov method is a reliable treatment for humeral nonunions, even after multiple previous operations or in the event of infection. PMID- 9728179 TI - Effect of nicotine on the rate and strength of long bone fracture healing. AB - Empirical clinical observation suggests that cigarette smoking had an inhibitory effect on long bone fracture healing, but this has not been proven scientifically. Forty female New Zealand White rabbits had midshaft tibial osteotomies performed and plated. These were divided randomly into two groups receiving either systemic nicotine or saline (placebo). Lateral radiographs were taken at 4, 6, and 8 weeks that showed a 17.2% average difference in callus formation between the two groups and a significant lag in formation of cortical continuity in the nicotine group. The rabbits were sacrificed 8 weeks after fracture, and healing was compared biomechanically. Three (13%) fractures showed no clinical evidence of union in the nicotine group, whereas all fractures in the control group healed. Biomechanical testing showed the nicotine exposed bones to be 26% weaker in three-point bending than were those exposed to placebo. PMID- 9728180 TI - Mechanical properties of perforated and partially demineralized bone grafts. AB - Changes in flexural rigidity and compression strength of 18 sheep tibias were investigated after laser perforation and partial demineralization. Test bones were divided into three groups: Group 1, no treatment; Group 2, laser hole grid; and Group 3, laser hole grid and partial demineralization. Starting in the anterior direction at the tibial tuberosity, the flexural rigidity was determined using a nondestructive four-point bending test. The elliptical distribution of the flexural rigidity before and after a specific treatment was compared. After the bending test, a cylindrical center section of each test bone was loaded axially to failure to determine subsequent changes in compression strength. Results showed that perforation alone produced minimal reduction of rigidity and insignificant changes in compression strength. However, additional partial demineralization resulted in larger reductions. In compression testing, perforated and partially demineralized bone specimen showed marked decrease of the ultimate failure stress. The observed increase in failure strain appeared to be related to compression of the laser holes. The findings of this study suggest that partial demineralization and perforation can be applied to diaphyseal bone grafts and that their decreased mechanical properties are a function of the bone volume reductions produced by both processes. PMID- 9728181 TI - Tensile load and the metabolism of anterior cruciate ligament cells. AB - It generally is recognized that tensile load plays a major role in maintaining the homeostasis of the anterior cruciate ligament fibers, but its detailed mechanism remains a matter of controversy. The effect of cyclic tensile load on the metabolism of the anterior cruciate ligament were investigated experimentally using cultured cells from the anterior cruciate ligament of rabbits. Using culture plates with flexible rubber bases, a cyclic tensile load was applied to the cultured cells for 24 hours, and the changes in shape, alignment, and metabolism of the cells were analyzed. Under the cyclic tensile load, the shape of the cells from the anterior cruciate ligament changed to spindle and aligned perpendicularly to the direction of the tensile load. The cyclic tensile load also caused an increase in collagen synthesis by the cells from the anterior cruciate ligament, which was predominant in Type I. The cells from the synovium showed similar changes in shape and alignment under the cyclic tensile load, but no significant change was observed in cell metabolism. These observations suggest that the application of cyclic tensile load on the anterior cruciate ligament cells is an important factor in the regulation of collagen synthesis in the anterior cruciate ligament. PMID- 9728182 TI - Sternal mass in an 11-year-old boy. PMID- 9728183 TI - Fluid therapy for the pediatric surgical patient. AB - The fluid management of the pediatric surgical patient is a crucial aspect of surgical care. This article reviews the fundamental physiology of fluid replacement in children and highlights how standard formulas for fluid therapy can be modified to account for the rapidly changing physiology of the pediatric surgical patient. Novel approaches to fluid treatment of the surgical patient with oral rehydration formulas are discussed. Finally, guidelines for specific management of common pediatric surgical diseases are presented. PMID- 9728184 TI - The approach to common abdominal diagnosis in infants and children. AB - This article focuses on salient points in the evaluation of abdominal pain in infants and children. Specifically, the authors address appendicitis and abdominal pain associated with either vomiting, constipation, or gastrointestinal bleeding. A discussion of common abdominal masses, urologic, and gynecologic problems, and considerations in the evaluation of immunologically suppressed or neurologically impaired children, and children with recurrent abdominal pain is also presented. The authors establish logical, focused approaches to the initial evaluation and management of abdominal pain and suggest criteria for timely surgical referral. PMID- 9728185 TI - Pediatric hernias and hydroceles. AB - Hernias and hydroceles are common conditions of infancy and childhood, and inguinal hernia repair is one of the most frequently performed pediatric surgical operations. As a result of improved neonatal intensive care, more and more premature babies are being delivered, and consequently the incidence of neonatal inguinal hernia is increasing. The most important aspect of the management of neonatal inguinal hernias relate to its risk on incarceration, and emphasis is placed on this point. This article covers the embryology, incidence, clinical presentation, and treatment of groin hernias and hydroceles, as well as dealing with abdominal wall hernias other than umbilical hernias. This article places special emphasis on when a patient with a hernia or hydrocele should be referred to a pediatric surgeon. PMID- 9728186 TI - Pediatric umbilical problems. AB - After birth, the normal umbilicus is a relatively simple structure. During the development of the embryo, however, this region is highly complex. Vestigial of the umbilical cord can be responsible for umbilical inflammation and drainage. This article reviews the embryology of the umbilicus and discusses a number of clinical problems seen in this area. The authors' aim is to aid the primary care pediatrician in evaluating, treating, and appropriately referring umbilical problems encountered in office practice. PMID- 9728187 TI - Circumcision and pediatric disorders of the penis. AB - Circumcision is a commonly performed procedure, but medical indications remain controversial. Most disorders of the penis in childhood can be diagnosed and managed by the primary care pediatrician. However, some require early recognition and surgical intervention, and prompt referral to a pediatric surgeon or urologist will optimize outcome. This article discusses the recognition and initial management of these problems, particularly those most commonly seen or followed in the outpatient setting. In addition, the authors review the issues of circumcision. PMID- 9728188 TI - Pediatric testicular problems. AB - Testicular problems in children may be both congenital and acquired. These problems are often difficult to diagnose and carry significant sequelae if untreated. Early surgical consultation is often needed for correction of the problem. This article reviews the pathophysiology of the most common pediatric testicular abnormalities with emphasis on the diagnostic modalities employed and current treatment alternatives. PMID- 9728189 TI - Minor pediatric injuries. AB - Injuries are a common source of childhood morbidity and mortality. The initial evaluation should follow in a sequential fashion to determine the extent of injuries. Most minor injuries can be treated safely and cost-effectively in an office setting. The principles of wound care include adequate hemostasis, tissue debridement, removal of imbedded foreign bodies, and appropriate closure or coverage of the wound to optimize healing. Appropriate use of antibodies, tetanus prophylaxis, and rabies immunization will minimize complications. With proper selection and treatment, the outcome of children with minor injuries should be excellent. PMID- 9728190 TI - Late sequelae of major trauma in children. AB - Traumatic injuries represent the most important threat to the health of children in the United States and are the leading cause of death after the first year of life. More than 20,000 children and young adults will die this year as a result of injury. For every child that dies, another 40 will require hospitalization and 1000 more will be evaluated in emergency departments. Moreover, 50,000 children and adolescents will sustain some degree of permanent disability, the majority of victims of brain injury. This article focuses on several important long-term implications for children, their families and primary care physicians, following multisystem injuries. PMID- 9728191 TI - Pediatric chest lesions. AB - Pediatric chest lesions are usually either symptomatic or strikingly visible. The most common lesions, as well as the lesions that require prompt surgical treatment, are reviewed in this article. Careful imaging studies and diagnostic tests such as bronchoscopy can usually characterize these lesions and enable a safe, directed surgical approach. Although many chest lesions can be managed without surgery, primary care providers can expedite treatment of these problems by early referral to pediatric surgeons or pediatric thoracic surgeons. PMID- 9728192 TI - Lymphadenopathy in children. AB - Lymphadenopathy is a common problem in children and adolescents. A detailed history and physical examination in addition to knowledge of lymph node anatomy is often adequate for diagnosis. The infectious and noninfectious causes of adenopathy are outlined according to location. Medical and surgical evaluation and treatment are discussed, with special attention given to mycobacterial infections, cat scratch disease, and lymphoma. PMID- 9728193 TI - Pediatric head and neck lesions. AB - Commonly encountered head and neck lesions in children are described with an emphasis on evaluation, diagnosis, and treatment. Congenital lesions typically require excision, although hemangiomas usually resolve spontaneously. Acute suppurative lymphadenitis is common and readily diagnosed. Chronic lymphardenitis remains a diagnostic challenge and must be differentiated from malignancy. Lesions that do not respond to antibiotics should be biopsied to exclude neoplasms. PMID- 9728194 TI - Caring for the former pediatric cardiac surgery patient. AB - Because of marked improvements in the early diagnosis and management of patients with congenital heart defects as well as the dramatic increases in surgical survival for patients undergoing correction of these defects, a large and growing population of survivors of congenital heart surgery present themselves for care to primary care pediatricians. This article highlights the need for primary care pediatricians to understand the common clinical problems they will see in this group of patients and what surgical strategies are used in the more complex defects. PMID- 9728195 TI - Outpatient pediatric orthopedics. Common and important conditions. AB - Many common pediatric orthopedic conditions can be managed by the pediatrician who has a knowledge of the natural history of these conditions. An accurate diagnosis is necessary to provide proper treatment, give advice to patients, or make referrals to the proper specialist. The authors' find that in approximately 95% of cases, a specific diagnosis can be made and that 40% of patient referrals for orthopedic problems could have been managed by the primary care physician. This article discusses some of the more common pediatric orthopedic problems often encountered by primary care physicians. PMID- 9728196 TI - Common topics in pediatric otolaryngology. AB - This article outlines the perioperative and long-term management of children undergoing tonsillectomy, adenoidectomy, tympanostomy with tube insertion, and sinus surgery. Indications for complications of each of the procedures is reviewed in this article. The authors also cover the management of children with tracheostomies. Several questions frequently asked by parents of children with a tracheostomy are addressed. PMID- 9728197 TI - Common ophthalmologic concerns in infants and children. AB - Pediatric ophthalmology differs from adult eye care in many aspects. Some disorders are seen only in children although others may be found in adults as well. A major difference between pediatric and adult ophthalmology is the impact that almost any disorder may have on the developing visual system. This article addresses common pediatric eye disorders and their potential effects on the visually impaired immature child. Referral guidelines and vision screening are also discussed. PMID- 9728198 TI - The noninvasive vascular laboratory. AB - The noninvasive vascular laboratory is an important resource in the management of patients with peripheral vascular disease. The commonly accepted uses of the vascular laboratory, including both indirect testing (segmental limb pressures, waveform analysis, digital pressures, exercise testing, reactive hyperemia) and direct vascular testing with CDI, have been reviewed. These techniques have changed how these patients are diagnosed, with less need for invasive diagnostic techniques. Patient management and intervention have also been altered by these advances. The use of noninvasive techniques and continue progress in this area should continue to advance patient care. PMID- 9728199 TI - Magnetic resonance angiography of lower-extremity arterial disease. AB - Improvements in vascular technique have expanded the treatment options for patients with severe occlusive peripheral vascular disease. The decision to perform a major revascularization procedure in patients who are often at high risk for cardiovascular morbidity and mortality depends on the risk-benefit ratio. Detailed and accurate vascular imaging is essential and evaluating the likelihood of a successful revascularization with subsequent limb salvage. Although contrast angiography has been the time-honored reference standard imaging technique, the method is an invasive procedure with limitations and risks. MRA is a new, noninvasive vascular imaging technique that may now be added to the imaging options with the potential for improved sensitivity for finding patent runoff vessels, avoidance of morbidity, and cost equivalent to that of conventional contrast angiography. Magnetic resonance angiography is a rapidly developing and exciting new vascular imaging technique. As with any new technique, it is imperative that individual centers validate their MRA results and interpretations against the time-honored standard, which continues to be contrast arteriography. Several studies now indicate that MRA can be a cost effective outpatient imaging technique sufficient for planning and successfully performing peripheral bypass procedures. As developments in hardware, software, and non-nephrotoxic contrast agents continue to increase, applicability of MRA in vascular surgery will continue to expand. Predictably, MRA will have a major role in the future of vascular imaging, and it is likely to supplant the need for conventional contrast angiography in the majority of patients. PMID- 9728200 TI - Diagnostic and interventional angioscopy. AB - As an imaging modality, angioscopy provides a simple method for the careful evaluation and treatment of the lumen of native vessels and bypass grafts. When used as a diagnostic study, angioscopy can provide more accurate information regarding the flow surface than conventional tests, such as angiography or duplex imaging. It can significantly enhance the ability of the surgeon to detect flow surface problems. With the recent advance in endovascular tools, angioscopically guided luminal intervention has become an increasingly useful approach to many vascular problems. More precise treatment of endoluminal abnormalities and a reduction in incision length and soft-tissue dissection can result in decreased patient morbidity and extended patient benefit. PMID- 9728201 TI - Intravascular ultrasound. AB - Vascular anatomy can be more accurately assessed from an endoluminal position applying the principles of conventional ultrasound. It may effectively batter select patients for endovascular procedures and complement the information gained from CT, MRA, transcutaneous ultrasound, and contrast angiography. Three dimensional and longitudinal grey-scale reconstruction of tomographic IVUS images are made available through computer software programs. Information gained includes: plaque characterization and morphology, a cross-sectional evaluation of stenoses, assessment of the impact/effect of balloon angioplasty, evaluation of vessel dissections and the ability to selectively place intravascular stents. PMID- 9728202 TI - Angiography with carbon dioxide (CO2). AB - CO2 possesses many advantages over conventional iodinated contrast agents used for arteriography. It is nonallergic and lacks renal toxicity. Its unique properties permit use of smaller catheters in diagnostic and therapeutic angiographic procedures, allow optimal vascular imaging of various neoplasm, assist in detection of occult gastrointestinal bleeding, and facilitate TIPS procedures. With digital subtraction techniques and stacking programs, CO2 arteriography is as accurate as iodinated contrast studies in most patients and thus is the preferred arterial imaging technique in patients with contrast allergy and renal insufficiency. CO2 is also extremely inexpensive compared with available contrast agents. Understanding of the effects of buoyancy and compressibility is necessary for safe, controlled delivery of CO2 during arteriography, but only rare complications have occurred in our large experience with CO2 angiography. Thus, use of CO2 as an arterial contrast agent significantly expands the safety and utility of arterial imaging in patients with peripheral vascular disease. PMID- 9728203 TI - Balloon angioplasty, stenting, and role of atherectomy. AB - Endovascular interventions for the treatment of lower-extremity atherosclerotic disease have undergone a rapid course of development during the past 30 years. Balloon angioplasty is the most widely applied of these techniques and has been shown to yield excellent results, at least in the short and intermediate terms, in appropriately selected patients. The ideal candidate is one with a focal stenotic lesion of the iliac artery; this is also the type of patient in whom placement of an endoluminal stent, whether primarily or as an adjunct to balloon angioplasty, has been shown to be most effective. The initial enthusiasm for transluminal atherectomy of lower-extremity atherosclerotic lesions has met with some disappointing long-term results. It is now used mainly in conjunction with either or both of the above modalities in a select group of patients. Clearly, more controlled studies of all these techniques are needed to better define their exact indications and limitations in treatment of lower-extremity arterial disease. PMID- 9728204 TI - Lower-extremity arterial endovascular stenting. AB - Intraluminal arterial stenting for the management of arterial occlusive disease of the lower extremities has evolved over the years. Most stents are used to correct inadequacies of PTA or to correct a PTA complication. These include (1) restenosis within 90 days of PTA, (2) chronic iliac occlusion, (3) acute occlusions during PTA, (4) a significant residual gradient following PTA, (5) dissections longer than the angioplasty site, and (6) a 30% or greater residual stenosis after PTA. Both the Palmaz stent and the Wallstent have performed well in the iliac artery system. Their use in the femoropopliteal system has not been as successful and should be reserved for selected cases. Long-term anticoagulation is generally required for femoropopliteal stent patency. Placement in the lower superficial femoral or popliteal artery is best avoided. Re-angioplasty or additional stenting may be used to prolong patency, although with the risk of a second intervention. Progression of arteriosclerosis is a factor to consider when choosing an endoluminal treatment versus standard bypass. PMID- 9728205 TI - Varying strategies for endovascular repair of abdominal an iliac artery aneurysms. AB - Endovascular grafting for abdominal aortic aneurysm has evolved over the past 7 years from Parodi's simple tubular aortic endografts with uniform attachment systems to more complex "modular" designs that begin to provide some flexibility of graft construction at the time of implantation. The future of these systems likely resides in a series of devices that contain sufficient flexibility to permit the operator the option for in situ customization of graft dimensions in accordance with realtime operative/fluoroscopic findings. PMID- 9728206 TI - Clinical applications of thrombolytic therapy for arterial and graft occlusion. AB - Catheter-directed thrombolysis and intraoperative intra-arterial thrombolysis are important adjuncts to how we care for patients with acute arterial and bypass graft occlusions. Of importance is that intra-arterial thrombolysis not to be thought of as a competitor to operative revascularization, but rather as an adjunct to what can be accomplished, which enables the responsible physician to offer the best care for these patients. PMID- 9728207 TI - New tumor markers of testis cancer. AB - Potential tumor markers for testis cancer have become numerous with the new molecular techniques available. New protein markers have been evaluated, and histologic factors have shown correlations with stage of disease. Cytogenetic analysis studies have also shown associations with stage progression. Chromosomal markers, oncogenes, and tumor suppressor genes are possible candidates for tumor markers. These new potential tumor markers may become as commonplace as the established markers and may enhance diagnosis, staging, and treatment of testis cancer. PMID- 9728208 TI - Imaging and management of atypical testicular masses. AB - Most testicular masses are germ cell malignancies and require radical orchiectomy. There are other causes of testicular masses, however, some of which have characteristic imaging and clinical features. A presumptive diagnosis may be possible for some of these atypical testicular masses. This may result in testis preserving surgery or nonoperative management. PMID- 9728209 TI - Risk assessment for metastatic testis cancer. AB - Metastatic germ cell tumors represent a model for curable malignancy, with 70% to 80% of patients cured of their disease. Patients with a good prognosis are likely to be cured, and the aim of investigation is to maintain the high cure rate while minimizing toxicity. For patients who fail to achieve a complete response (CR) to therapy, or who relapse after achieving CR, the prognosis is poor--these patients have become the focus of more intensive treatment to increase the cure rate. Distinguishing which patients will require more aggressive therapy ab initio has been the goal of successive attempts to stratify patients into either good- or poor-risk groups to tailor treatment accordingly. A number of analyses have yielded variable factors and differing schemas to classify patients, culminating in the International Germ Cell Consensus Group whose findings have been utilized to develop the new testicular cancer staging system for the American Joint Committee on Cancer (AJCC) and the Union International contre le Cancre (UICC). This article traces the developments in risk assessment that have lead to this consensus, which would prospectively allow comparison among future clinical trials. PMID- 9728210 TI - Staging of testis cancer. Combining serum markers, histologic parameters, and radiographic imaging. AB - Treatment of testis cancer has improved dramatically over the last 25 years with cure rates that now approach 95%. This success in treatment is the result of multimodal therapy that may include cisplatinum-based chemotherapy, surgery, and radiotherapy. Although there is little dispute that chemotherapy is appropriate as initial treatment for advanced testis cancer, controversy remains regarding the management of early stage B and stage A testis cancer. In part, the current controversy is driven by the performance characteristics of presently available staging methods, that may include serum markers, histologic parameters from orchietomy specimens, and radiographic imaging. This article reviews the performance characteristics and use of these staging methods. PMID- 9728211 TI - A rational approach to managing stage I nonseminomatous germ cell cancer. AB - Regardless of the treatment option selected for management of low-stage germ cell cancer, ultimate survival is nearly identical. Treatment-related morbidity is very low regardless of management modality and the individual patient can expect similar physical limitations owing to therapy. The overall difference in loss of productivity between treatment programs varies by little more than 1 week. The cost of treatment is similar for all methods, although there is a definite financial advantage to surveillance, less so for selective surveillance, when compared with other forms of management. Socioeconomic factors are of importance when managing limited resources for a large population, but are of less concern to an individual, especially when the mean differences in per patient costs vary by only $5000. Because of these close similarities in efficacy, morbidity, and costs treatment decisions should be individualized. A responsible and reliable patient can be managed safely by selective surveillance. Those individuals considered to be less self-motivated to pursue intensive care should be managed by primary therapy. Without more information regarding the long-term outcomes associated with primary adjuvant chemotherapy, primary adjuvant RPLND, where experienced surgical support is available, is the preferred management for low stage germ cell cancer in patients selected for, or electing, active treatment rather than surveillance. Active investigations examining the role of medical management in this population should be continued. Our preferred choice of initial management is to offer selective surveillance to appropriate patients and modified RPLND to the remainder. PMID- 9728212 TI - Surveillance for stage I testicular seminoma. Is it a good option? AB - Postorchidectomy treatment options in patients with stage I seminoma include surveillance (reserving treatment for patients who relapse), adjuvant radiation therapy (RT), and adjuvant chemotherapy. Adjuvant retroperitoneal RT remains the treatment of choice in most centers; however, the success of surveillance in stage I nonseminomatous germ cell testis tumors, the establishment of curative chemotherapy for advanced disease, and the improvements in CT have led to re examination of the standard treatment approach. The available data from the surveillance and adjuvant RT series suggest that almost 100% of patients with stage I testicular seminoma are cured, whichever approach is chosen. This article presents an overview of the available information on all treatment options, the pros and cons of each approach, and indications for where surveillance fits into the armamentarium of clinicians dealing with this disease. PMID- 9728213 TI - The management of stage I testis cancer. AB - For clinical stage I seminoma, conventional management consists of adjuvant RT after orchiectomy. Only 5% of patients relapse. The majority can be salvaged by chemotherapy. The overall survival of 98% is excellent. Seminoma is radiosensitive. A lower dose of RT is required than for NSGCT. Standard therapy presently is 30 Gy in 3 weeks, as suggested by the MRC study. RT is generally well tolerated. There have been recent concerns about second malignancies after 10 to 15 years. Surveillance studies have shown that 18% of patients relapse, the majority in para-aortic lymph nodes. About 15% require salvage RT and 5% salvage chemotherapy. Second relapses are seen in patients treated with RT at first relapse, and occur outside of the radiation field. The main advantage of surveillance is that 80% of patients can be spared slightly toxic overtreatment. The main disadvantage is the need for long-term follow-up, which is expensive and stressful to the patient. Good patient compliance, mandatory to an observation policy, is often difficult on a long-term basis. Seminoma is clearly responsive to chemotherapy. Adjuvant carboplatin in clinical stage I has only been evaluated in two studies. Because reliable prognostic factors have not been established, a high-risk group cannot be identified, and chemotherapy must be given to all patients. Whether or not one cycle of chemotherapy is sufficient requires further confirmation, particularly in view of the results with carboplatin as compared with cisplatin in patients with advanced NSGCT. Results of the randomized MRC trial comparing RT with carboplatin are of interest. PMID- 9728214 TI - Stage II nonseminomatous testis cancer: the roles of primary and adjuvant chemotherapy. AB - Historically, a retroperitoneal lymph node dissection (RPLND) has been the primary treatment of stage II nonseminomatous germ cell tumor (NSGCT) patients, whereas chemotherapy has been used in the adjuvant setting. Increasing evidence of the efficacy of chemotherapy has led to additional treatment possibilities for stage II patients. Both chemotherapy and RPLND have a role in the primary therapy of stage II NSGCT, and both modalities can be used as secondary adjunctive therapy. Management of these patients now requires a renewed emphasis on risk stratification and on the integration of patient preferences into the management algorithm. PMID- 9728215 TI - Retroperitoneal lymphadenectomy in staging and treatment. The development of nerve-sparing techniques. AB - Currently, where available and widely practiced, nerve-sparing RPLND offers the least toxic and safest alternative in management of clinical stage I nerve sparing germ cell tumor. Twenty-six percent to 30% of such cases are found to be node positive and, if no adjuvant chemotherapy is elected, two thirds do not relapse; those who do are uniformly rescued by chemotherapy. The 70% who are node negative have no long-term toxicity and do better in quality of life studies than their surveillance counterparts. Also, risk-benefit and cost-benefit studies support the competitive position, of RPLND vis-a-vis the other options. Nonetheless, it is fair to say there are several options for management of clinical stage I nonseminomatous germ cell tumors that, when well applied, work well. Their choice depends on regional factors related to availability and experience. PMID- 9728216 TI - Laparoscopic retroperitoneal lymphadenectomy for cancer of the testis. AB - Laparoscopic retroperitoneal lymph node dissection (RPLND) is a technically advanced procedure that has been undertaken for the management of low-stage nonseminomatous germ cell testis tumor. Although it has been shown to be an effective staging technique, its role as a therapeutic operation is currently unknown. Laparoscopic RPLND requires longer operative times but offers the patient all the advantages of minimally invasive surgery, such as less postoperative pain and shorter hospitalization and convalescence. The role of laparoscopic RPLND for the evaluation of residual abdominal masses following chemotherapy is currently being examined. PMID- 9728217 TI - Can retroperitoneal lymphadenectomy be omitted in some patients after chemotherapy? AB - The advances in the therapy of high-volume stage II and stage III testicular cancer have been largely due to the development and success of cisplatin-based chemotherapy. Though patients with low- to moderate-volume stage II disease historically were curable with radical retroperitoneal lymph node dissection, patients with higher volumes of metastatic disease were not curable with surgery alone. The advent of cisplatin-based chemotherapy in the 1970s has allowed many of these patients with higher-volume metastatic disease to be cured. Though chemotherapy clearly plays the major role in the therapy of these patients, surgery after chemotherapy has been complimentary in the overall chance for cure of these patients. PMID- 9728218 TI - Poor-risk germ cell tumors. Recent developments. AB - Standard therapy for patients with poor-risk germ cell tumors remains four cycles of bleomycin, etoposide, and cisplatin (BEP) chemotherapy, which cures approximately 50% of this patient population. Randomized trials of conventional dose chemotherapy have failed to identify a regimen with superior efficacy to BEP. Preliminary data suggesting efficacy for high-dose chemotherapy with stem cell or autologous bone marrow transplantation as initial therapy for poor-risk disease have led to two randomized trials of this approach as initial therapy for poor-risk patients. The recently developed International Germ Cell Consensus Classification provides a standard method for determining poor-risk status and should be uniformly applied for both clinical decision making and risk assignment in clinical trials. PMID- 9728219 TI - Cost-effective strategies for the follow-up of patients with germ cell tumors. AB - Cost-effective strategies for the follow-up of patients with germ cell tumors must be based on the known natural history of the disease, the accuracy and cost of diagnostic modalities, and the efficacy and effectiveness of therapy when recurrences are detected. The natural history of stage I and stage II germ cell tumors are reviewed, including the unique circumstances of late recurrences. The accuracy and cost of imaging modalities are also reviewed and general recommendations to cost-effective follow-up are proposed. PMID- 9728220 TI - Long-term side effects of treatment for testis cancer. AB - Patients newly diagnosed with testis cancer can now expect excellent results with respect to long-term, disease-free survival after treatment. Given the young age of presentation for many of these patients, the long-term consequences of curing testis cancer have become a major concern. Surgery, radiation, and chemotherapy for testis cancer have all been associated with potential long-term side-effects. Consequently, new treatment regimens have been directed toward minimizing these possible side-effects while at the same time maintaining high cure rates (i.e., limiting the size of radiation fields, decreasing the number of chemotherapy cycles, eliminating bleomycin). Patients and physicians must be made aware of the potential adverse side effects of treatment for testis cancer. At the present time, however, it appears that the beneficial effects of such treatment, with respect to overall and disease-free survival, far outweigh the limited probability of persistent treatment-related side effects in patients newly diagnosed with testis cancer. PMID- 9728221 TI - Fertility issues and their management in men with testis cancer. AB - Although a curable malignancy, testis cancer and its treatment have unique associated morbidities that largely affect reproductive dysfunction. In this focused review, the factors that contribute to infertility in men with testis cancer are outlined. The treatment-specific risks to fertility that accompany cancer management are also discussed. Contemporary methods of overcoming infertility in testis cancer patients are addressed, and several exciting and promising experimental approaches to the preservation or restoration of fertility for men with testis cancer are presented. PMID- 9728222 TI - Laparoscopic bladder autoaugmentation in children. AB - Answering the need for a simple, nonenteric method of increasing bladder capacity, this article offers a new laparoscopic procedure for autoaugmentation of the bladder in children. An outline of the technical design is given. Experience with seven children is delineated. Postoperative voiding control and function capacity were greatly improved; this procedure is judged worthy of addition to the surgical armamentarium. PMID- 9728223 TI - Resting the lung. PMID- 9728224 TI - Chronic cough--mechanisms and management. PMID- 9728225 TI - The investigation and treatment of sleep-related breathing disorders. PMID- 9728226 TI - The mechanism of action of corticosteroids in asthma. PMID- 9728227 TI - Toxicology of the lung. PMID- 9728228 TI - Systemic inflammatory responses to cardiopulmonary bypass: a pilot study of the effects of pentoxifylline. AB - The potential of pentoxifylline (PTX) to modify systemic inflammatory responses and lung injury following cardiopulmonary bypass (CPB) was studied in 20 patients undergoing elective coronary artery surgery. Ten control patients were compared with ten patients who received a PTX infusion of 1 mg kg-1 h-1 during surgery. Intra-vascular pulmonary leukocyte sequestration was observed in neither group following discontinuation of CPB. Plasma elastase-alpha-1-antiprotease complex rose three-fold from baseline in both groups to peak at sternal closure. No significant plasma interleukin-1 (IL-1) response was detected. Plasma interleukin 6 (IL-6) rose in both groups from baseline to peak 4 h postoperatively. There was no correlation between plasma levels of elastase complex, IL-1 or IL-6 and impairment of postoperative oxygenation. CPB was associated with significant postoperative hypoxaemia and systemic release of neutrophil elastase and IL-6 but PTX, at the given dose, did not abrogate these responses. PMID- 9728230 TI - The prevalence of preferential nasal breathing in adults. AB - The prevalence of preferential nasal breathing was studied in an awake adult population. One hundred and ninety-four people consented to gentle manual compression of the nostrils. They were advised to 'breathe in and out', but no further information regarding breathing was given to avoid influencing the patient. One hundred and eighty patients (92.8%) commenced immediate regular relaxed breathing. Fourteen patients (7.2%) had difficulty with oral breathing which ranged from irregular mouth breathing associated with distress to no spontaneous respiration. The prevalence of preferential nasal breathing was strongly associated with increasing age (chi 2 for trend, P = 0.007). In addition, a weakly significant association was demonstrated between a history of asthma and this phenomenon (P = 0.047). These findings suggest a tendency for the elderly person to revert to the infant pattern of obligate nasal breathing. Physicians should be aware of this possibility in the elderly patient, especially prior to any procedure which may induce nasal obstruction. PMID- 9728229 TI - Early management of younger adults dying of community acquired pneumonia. AB - We identified all patients under the age of 65 dying from CAP over a 3-yr-period in hospital in our health district. Most had chronic underlying illnesses. The early management of those who had previously been in good health was studied in greater detail, and was not ideal. Recommendations have been made to try and improve the assessment and treatment of patients with severe community-acquired pneumonia. PMID- 9728231 TI - Sodium cromoglycate via inhaler and Autohaler. AB - One hundred and eighty-one children with asthma were entered into a randomized, controlled, crossover study comparing sodium cromoglycate via a metered dose inhaler (MDI) and a breath-actuated inhaler (Autohaler). There were no significant differences in pulmonary function tests (FEV1, FVC, PFER), symptom scores and bronchodilator use between the two devices. However both patients' and clinicians' opinions of sodium cromoglycate effectiveness were significantly better (P < 0.01) for the Autohaler. Autohaler was also thought to be easier to use (P < 0.001) and better for co-ordination of actuation with inhalation (P < 0.001). The Autohaler is as efficacious as the metered dose inhaler used by good co-ordinators. It seems to be the significantly better device with respect to ease of use, actuation and co-ordination which may aid compliance. PMID- 9728232 TI - Bacteriology of tonsil and adenoid and sampling techniques of adenoidal bacteriology. AB - The value of pernasal swabs and direct adenoid swabs in chronic adenoid and adenotonsillar disease was assessed in 175 patients. Prior to adenoidectomy (53 patients) or adenotonsillectomy (122 patients), pernasal and direct adenoid swabs were taken. Adenoid currettings and tonsil tissue were cultured. Haemophilus influenzae was the bacterium most frequently isolated from adenoid currettings and from the centre (core) of the resected tonsil. There was a close relationship between the bacteriology of the pernasal swab and the adenoid tissue and tonsil core in 72 and 71% of patients, respectively. There was an identical profile of pathogens in 52 and 49%, respectively. We suggest that in children with adenoiditis or adenotonsillitis and hypertrophy of the adenoid, a pernasal swab should be used in preference to a throat swab in selecting appropriate antimicrobial therapy. Penicillin and ampicillin are not appropriate blind therapy in chronic adenoid and adenotonsillar infections because of the prevalence of beta-lactamase-producing aerobes (40%) in adenoid and tonsil core in these conditions. PMID- 9728233 TI - Asbestos-related pericardial thickening detected by magnetic resonance imaging. AB - Magnetic resonance imaging was performed in four male asbestos workers in whom the chest radiograph revealed pleural but not pulmonary or pericardial disease. Patients underwent thoracic multislice spin echo imaging, with measurement of left and right ventricular volumes at end-diastole and end-systole, and a study of the flow in the superior vena cava as an indirect measure to the filling of the right ventricle. Patients also underwent respiratory function tests and high resolution computed tomography (HRCT). Magnetic resonance, but not HRCT, showed pericardial thickening in two patients. Magnetic resonance demonstrated reduced diastolic flow in the superior vena cava in one patient, reflecting impaired right ventricular filling. All other magnetic resonance measurements of cardiac function were normal. HRCT demonstrated mild asbestosis in three patients in which neither the chest radiograph nor magnetic resonance showed signs of parenchymal disease, and pericardiac calcification without thickening in one patient. It is concluded that magnetic resonance is superior to HRCT in identifying pericardial thickening, but that HRCT is superior to magnetic resonance in identifying asbestos-related pleural and pulmonary disease. PMID- 9728234 TI - Spontaneous pneumomediastinum in a patient with fibrosing alveolitis. PMID- 9728235 TI - Inflammatory basis of asthma. PMID- 9728236 TI - Evaluation of mean transit time. PMID- 9728237 TI - Phenomenon of rifampicin-induced discolouration of body fluids. PMID- 9728238 TI - Hypothalamo-pituitary-adrenal axis suppression in asthmatics. PMID- 9728240 TI - The acquired Chiari malformation and syringomyelia following spinal CSF drainage: a study of incidence and management. AB - Firstly, 14 patients are described who developed either an acquired Chiari malformation (ACM) alone (7 cases) or ACM and syringomyelia (7 cases) after lumbar subarachnoid space (SAS) shunting or in one case, epidural anaesthesia with SAS penetration. Four groups are considered: 3 cases with craniofacial dysostosis and communicating hydrocephalus (CH), 4 cases with CH alone, 3 cases with pseudotumour cerebri (PTC) and a miscellaneous group (4 cases). Initial treatment was varied: resiting the shunt to ventricle or cisterna magna [6], adding an H-V valve [1], syrinx shunting [4] and posterior fossa decompression [3]. Further treatment was required in 6 cases. Secondly, incidence was examined in 87 patients with PTC initially treated either by lumbar SAS shunting [70] or cisterna magna shunting [17]. In the first sub-group, 11 cases (15.7 per cent) developed an ACM, 3 symptomatic (as above) and eight asymptomatic with 1 case also having syringomyelia whereas 1 case occurred in the second group with a questionanably symptomatic ACM. While accurate for symptomatic lesions, these figures are tentative with respect to asymptomatic lesions due to inadequate pre treatment radiology and detailed MR follow-up. The main conclusions are, first, that the incidence of symptomatic ACM and/or syringomyelia is not high enough to warrant abandoning SAS shunting; second that asymptomatic lesions need not necessarily be treated and third, that when treatment is required, shunt resiting is the first choice. PMID- 9728239 TI - Vasa vasorum of the intracranial arteries. AB - Most of the major extracranial arteries have vasa vasorum which play an important role in some pathological conditions. However, in the intracranial arteries, the existence of vasa vasorum and their pathological implication have not been adequately investigated. We examined the distribution and incidence of vasa vasorum in the major cerebral arteries and their relationships to certain clinical factors in 50 autopsy cases performed between 1987 and 1994. By light microscopy, vasa vasorum were found in 36 of 50 patients. Of 36 patients, vasa vasorum in 30 cases were localizedly observed in the tunica adventitia and the in other 6 were distributed in the tunica media accompanied by intramural haemorrhage. Existence of vasa vasorum was more common in the proximal arteries (vertebral, internal carotid, and basilar arteries) than in the distal arteries (middle cerebral and anterior cerebral arteries). Vasa vasorum were found more frequently in aged patients with severe atherosclerosis and those with cerebrovascular diseases. Our results indicated that intracranial vasa vasorum existed with a higher frequency in the tunica adventitia of the vertebral and internal cerebral arteries, and the incidence of vasa vasorum related to severity of atherosclerosis. The development of vasa vasorum in the tunica media may reflect some pathological changes of cerebral arteries. PMID- 9728241 TI - Three-dimensional computerized tomography in the presurgical evaluation of Chiari malformations. AB - Malformations of the cranio-cervical junction represent a complex entity, comprising neuro-meningeal and bone anomalies. Malformations of the central nervous system are nowadays easily explored using magnetic resonance imaging (MRI), while osseous malformations are classically assessed with standard radiographic techniques, which can give only incomplete information, and are often difficult to realize and analyse. We report a retrospective study of 10 patients with a Chiari malformation operated on in our department at La Salpetriere hospital, between July 1995 and December 1996 (1 man, 9 women; mean of age: 35 years; postoperative median follow-up: 6 months; 70% of improvement and 20% of stabilisation), and we underline the interest of the systematic pre operative realization of a 3D CT and angio-CT studies of the cranio-cervical junction. This examination offers advantages of: a precise analysis of the more complex osseous malformations; a study of the relationships between vascular and bony structures; a study of the relationships between neuraxis and spinal and cranial structures; and an optimal planning of the surgical procedure, adapted to the anatomical particularities of the patient. PMID- 9728242 TI - Gamma-knife surgery for secreting pituitary adenomas. AB - We report our preliminary results concerning 25 patients with secreting pituitary adenomas treated with stereotactic radiosurgery after partial transsphenoidal surgery and followed over a 6-36 month-period. Among the 15 acromegalic patients, a decrease of 65% in mean GH levels was achieved after 6 months and of 77% at 12 months after radiosurgery. Presently, only 3 patients (20%) are considered as in remission (mean GH and IGF1 level into the normal range). A decrease of 46% and 64% was observed at 6 and 12 months after radiosurgery in 4 patients with prolactinomas although no normalization of PRL levels occurred. Presently, 3/4 patients have individual PRL level slightly above the normal range. A normalization of Urinary Free Cortisol (UFC) was noticed in 4/6 (66%) patients with Cushing's disease within 6-12 months. No pituitary deficiency was noticed in this series with the exception of 4/25 patients (16%) who received subtotal or total pituitary irradiation for post-operative remnants of secreting adenomas poorly defined on MRI. One woman, who had undergone previously a conventional irradiation and presenting with a cavernous sinus adenoma reaching the optic nerve, developed an optic neuropathy. A second woman, with a cavernous sinus remnant, presented a cranial nerve palsy (VI) after the irradiation. We can conclude that radiosurgery using the Cobalt-60 Gamma-unit is, at least, as effective as conventional radiotherapy in the control of pituitary hormone hypersecretion from postoperative adenomas remnants with less adverse effects. PMID- 9728243 TI - An adult case of cerebral primitive neuroectodermal tumour. PMID- 9728244 TI - Management of recurrent craniopharyngioma. AB - Although histologically benign, craniopharyngioma can regrow either from macroscopic remnants of the tumour left behind at operation, or even after an apparently gross total removal. Recurrence rates vary significantly in the literature, depending on the efficacy of surgical treatment and also on the growth potential of the tumour itself. The main factor influencing tumour regrowth is obviously the extent of surgical resection, as total removal carries a much lesser risk of recurrence compared to subtotal or partial resections (although in such cases radiation therapy can lower this risk significantly). Other factors involved are the duration of follow-up and patient's age at operation, as children tend to relapse more frequently than adults. Even in the "microsurgery" era, characterized by high percentages of total resections, recurrences remain high and continue to represent a major problem of craniopharyngioma treatment. Twenty-seven children and adolescents were operated on for craniopharyngioma at the Department of Neurosurgery, Section of Pediatric Neurosurgery, Catholic University Medical School, Rome between June 1985 and June 1997. Total tumour resection was achieved in 18 cases, subtotal in 7 and partial in 2 instances. One patient died post-operatively. Post-operative neuroradiological investigations confirmed the operative findings, although 3 children with an apparently gross total removal showed a residual non-enhancing calcium fleck adherent to the hypothalamus (which remained stable at the following examinations). Three of the 9 patients with less than total removal underwent post-operative radiation therapy. Out of the 26 surviving patients 6 presented a recurrence of their craniopharyngioma, 2 after an apparently gross total removal and 4 after a subtotal or partial resection (one of them had received radiation therapy). The diagnosis was merely neuroradiological in 5 cases, as only one child presented a clinical picture suggestive of tumour regrowth. Surgery was the first therapeutic option in all the cases. Total tumour resection was accomplished in 3 cases, subtotal in 2 and partial in the last one. One child died post-operatively. Four of the 5 survivors received radiation therapy. All the patients are presently alive and stable (mean follow-up: 5.6 yrs). The authors conclude that surgery should be the first therapeutic option in case of recurrent craniopharyngioma and that radiation therapy should also be considered but only as adjuvant therapy. PMID- 9728245 TI - Clinical features of moyamoya disease in sibling relations under 15 years of age. AB - This study was undertaken to define the clinical features of moyamoya disease in sibling relations less than 15 years of age. We analysed five pairs of siblings (6 boys, 4 girls) with moyamoya disease from among 56 moyamoya patients less than 15 years of age. Of 56 paediatric patients with moyamoya disease, 5 were sibling pairs. At onset of the disease, all patients were under 10 years of age. Clinical manifestations in the proband and the affected sibling tended to present as transient ischaemic attacks; none of the 10 patients presented with intracranical haemorrhage. The probands were not always the older sibling, however, the time lapse between disease onset in the proband and his/her sibling was less than one year. Among the sibling pairs, there was no striking difference in disease staging based on angiographic findings. The incidence of sibling occurrence of moyamoya disease appears to be higher than previously assumed and the family of children with moyamoya disease should be cautioned that their other children have an increased risk of developing the illness. PMID- 9728246 TI - A comparison between ventriculo-peritoneal and ventriculo-atrial cerebrospinal fluid shunts in relation to rate of revision and durability. AB - Results of 884 first-time shunts inserted in the time period from 1958 to 1989 are retrospectively evaluated, 1) to perform a durability analysis of a shunt based on Kaplan-Meyer method, 2) to compare the rate of revision for ventriculo atrial (VA) and ventriculo-peritoneal (VP) shunts, 3) to compare the durability of a VA shunt with a VP shunt and 4) to do a stratified durability analysis comparing the VA and VP shunts in relation to the following background variables: shunt type, time period and age of the patient. Furthermore the specific complications related to VA and VP shunts are identified based on findings in the literature. Overall one-year shunt durability is 57% and five-year shunt durability is 37%. The median shunt durability is 1.68 years. Revision rate is 51% for VA shunts and 38.5% for VP (p < 0.05). Shunt durability is longer for VP shunts though the difference is not significant (p < 0.1). By use of stratified analysis of shunt durability no differences however are found between the two shunting methods. Hence the apparent difference in revision rate between VA and VP shunts seems secondary to variations in follow-up time and variations in background variables. To supplement our statistical analysis we have performed a literature study to look at the specific complications associated with VA and VP shunts. It seems as if the specific complications in relation to the VA shunting method are more severe than in relation to the VP shunting method. PMID- 9728247 TI - Altered excitability of the motor cortex after minor head injury revealed by transcranial magnetic stimulation. AB - This study attempts to find out whether the motor evoked potential (MEP) elicited by single pulse and slow-rate (1 Hz) repetitive transcranial magnetic stimulation (TMS) can disclose concealed subclinical impairments in the cerebral motor system of patients with minor head injury. The motor response to single pulse TMS (STMS) of the patient group was characterized by significantly higher threshold compared with that of the control group. The central motor conduction time, as well as the peripheral conduction time were normal in all patients pointing to cortical impairment. Two main patterns of MEP changes in response to repetitive TMS (RTMS) were observed in the patient group. A.--progressive decrease of the MEP amplitude throughout the stimulation session to a near complete abolition. B.--irregularity of the amplitude and the waveform of the MEP in a chaotic form. The MEP latency remained stable during the whole stimulation session. The MEP abnormalities recovered gradually over the period of a few months. The higher threshold of the motor response to STMS and the abnormal patterns of the MEP to RTMS seem to reflect transient impairment of cortical excitability or "cortical fatigue" in patients who sustained minor head injures. Further study is needed to evaluated the extent and the pathophysiological mechanisms of the central nervous system fatigue phenomenon following head injury. PMID- 9728248 TI - Selective musculocutaneous fasciculotomy for spastic elbow in cerebral palsy: a preliminary study. AB - Three hundred and ten people with cerebral palsy who had spasticity in one or more limbs underwent selective motor fasciculotomy (SMF) of the nerves supplying the harmful spastic muscles with the aim of achieving useful tone and to improve voluntary movements. Among them, 52 people (average age 9.5 years) had 75 spastic elbows who were considered fit cases to undergo SMF of the musulocutaneous nerve (MCN). The nerve was dissected in the upper 1/3rd of the arm. Bipolar current was used to stimulate the component fascicles and to detect those carrying excessive impulses. Some of the hyperactive fascicles were ablated according to preoperative grading of the spasticity, etc. Total relief in spasticity was achieved in 47 (62.66%) elbows. Whereas, in the remaining 28 (37.33%) elbows some degree of spasticity persisted. There were overall beneficial effects of SMF on the motor functions and the flexed elbow posture. There were no side effects and recurrence of spasticity. The results were observed for an average period of 17 months. It must be noted that, 5 people who had involuntary elbow flexion on activity, like walking, also developed normal posture and the to & fro swinging movements following surgery. In conclusion, SMF of MCN is an effective and safe procedure for achieving longlasting useful tone and voluntary movements in the harmful spastic elbow of people with cerebral palsy. The present report is an account of the largest number of cerebral palsy people in the world literature to date. PMID- 9728249 TI - Trepanation is a metaphor. PMID- 9728250 TI - Belgian doctors' attitudes on the management of patients in persistent vegetative state (PVS): ethical and regulatory aspects. AB - BACKGROUND: The best care and management of patients in persistent vegetative state (PVS) has been the subject of sustained moral and legal debate for a number of years. However, the views of specialist doctors in Belgium involved in the care for patients in PVS are largely unknown. METHODS: A postal questionnaire was sent to 403 members of Belgian Societies of Neurosurgeons, Neurologists and Rehabilitation Doctors. Their views were sought on various aspects of the management and care of PVS, focusing on the issue of the appropriateness of non treatment and the withdrawal of artificial feeding. FINDINGS: Of the 208 doctors who completed the questionnaires (52%), 172 (83%) indicated that they had been involved in the management of a patient in PVS. 88% of the responding doctors thought it was sometimes appropriate not to treat acute infections or other life threatening conditions in a PVS patient. Fifty-six percent considered it sometimes appropriate to withdraw artificial feeding. About three-quarter of physicians who considered both treatment-limiting decisions appropriate thought that such decisions could be considered within the first year of the patient being in PVS. Forty percent accorded a decisive influence to an advance directive and only a small number of doctors considered the influence of the patient's family in the decision to withdraw artificial feeding as decisive. Over 80% of the clinicians disagreed with the view that each decision about withdrawing artificial nutrition and hydration (ANH) should come before the courts. INTERPRETATION: Doctors in Belgium seem to be more reluctant to withdraw artificial feeding than not to treat acute infections or other life-threatening conditions in PVS. The reason for this difference appeared to be connected with the moral as well as with the clinical content of the decision. The broad variety of answers on the interval when the vegetative state is to be regarded as permanent and when treatment-limiting decisions are appropriate, could be due to the lack of official guidelines in Belgium. There seems however to be no consensus about a future policy in Belgium for making decisions about the withdrawal of ANH. PMID- 9728251 TI - Sutureless repair of major intracranial vessels with the Sundt clip-graft: technical note. AB - Major intracranial vessels can be damaged during tumor resection. With the availability of refined microvascular techniques, direct repair or by-pass of the damaged segment is possible. These methods, however, often require temporary occlusion of the offending vessel, can result in a less than optimal angiographic result, and are difficult to perform in a deep field. Additionally, in some patients direct repair or by-pass is not feasible because of the friability of the vessel or as a result of the large size of the tear. In these cases the Sundt clip-graft represents a valid adjunct to the armamentarium of the surgeon. Over the years, it has been used by the senior author in five patients where vascular injury occurred during the removal of brain tumors (3 meningiomas, one pituitary adenoma, and one low-grade glioma). In this report we illustrate our most recent experience with this ingenious tool. A 22-year-old man underwent resection of a recurrent left temporal lobe low-grade glioma. During resection of the tumor, a tear occurred in a branch of the middle cerebral artery. The tear was repaired using a Sundt clip-graft. A post-operative angiogram, performed five days later, showed patency of the vessel with no evidence of wall irregularities. Described 30 years age to be used primarily in aneurysm surgery, the Sundt clip-graft provides an excellent, too often forgotten, sutureless method of repairing intracranial vessels damaged during tumor removal. PMID- 9728252 TI - Damage to tumour and brain by interstitial photodynamic therapy in the 9L rat tumour model comparing intravenous and intratumoral administration of the photosensitiser. AB - In the 9L rat brain tumour model the damage to tumour and normal brain by photodynamic therapy after intratumoural photosensitizer administration (intratumoural PDT) was studied. Twenty four rats received an intratumoural injection of 4 or 40 mm3 haematoporphyrin derivative (HpD, 5 mg ml-1), followed by interstitial irradiation with 20 Joule (J) (630 nm) 5 h later. For comparison, seven rats were treated with 20 Joule 24 h after an intravenous injection of 10 mg kg-1 HpD (intravenous PDT). With the chosen PDT parameters there was no important difference between the damaged areas produced by intratumoural PDT or intravenous PDT. No selective tumour kill was observed. Even though normal brain tissue was heavily damaged, vital tumour parts were still present. Intravenous PDT caused extensive diffuse damage to small blood vessels in tumour and surrounding normal brain. Intratumoural PDT was characterised by an infiltration of polymorphonuclear cells into damaged tissue, dilatation of larger blood vessels and gross haemorrhage. These results suggest an immediate vascular shutdown in the intravenous approach, while in the intratumoural approach the vasculature remained patent initially. Because of the severe side effects observed, the use of HpD seems not advisable for intratumoural PDT of brain tumours. PMID- 9728253 TI - 5-Aminolevulinic acid induced endogenous porphyrin fluorescence in 9L and C6 brain tumours and in the normal rat brain. AB - A new approach in photodynamic therapy is the use of endogenous porphyrins for sensitisation of tumours to light. The induction of endogenous porphyrins after intravenous injection of 5-aminolevulinic acid (ALA, 200 mg kg-1) was studied in 23 rats, bearing intracranial 9L or C6 tumours. After 0, 2, 4, 6, 8, and 22 hours the rats were sacrificed and the fluorescence distribution of endogenous porphyrins was studied in brain tissue sections with a standard fluorescence microscope and a confocal laser scanning microscope. The role of blood-brain barrier disruption on porphyrin production was studied in 2 rats with a cryo lesion of the cortex. Additionally, 9L and C6 tumour cell cultures were incubated with ALA for 8 hours in vitro. Fluorescence was measured with a fluorescence spectrophotometer in cell cultures and in the brain sections. Porphyrins were detected in vitro in the tumour cells from 2 hours onwards and ex vivo in the tumour sections mainly from 2 to 8 hours, by 22 hours porphyrin fluorescence had almost disappeared. The contralateral brain showed low fluorescence levels between 2 and 6 hours after ALA administration. At the site of the cryo-lesions low fluorescence was measured 6 hours after ALA administration. The 9L tumours fluoresced homogeneously, with a sharp demarcation towards normal brain tissue. Fluorescence in the C6 tumours was patchy, with a poorly fluorescing edge. In both tumour models fluorescence was also detected in brain surrounding the tumour and sometimes in contralateral white matter and ventricle ependyma and pia mater. The slight increase of porphyrin fluorescence in the normal brain of tumour bearing rats, compared to the absence of this in rats without a tumour, was attributed to transport by bulk flow of porphyrins made in the tumours, and possibly also of circulating porphyrins or ALA leaking from the tumour vessels. PMID- 9728254 TI - Misjudged Bannwarth's syndrome culminating in laminectomy. PMID- 9728255 TI - Detection of a newly-developed thin-walled out pouching ("bleb") in an unruptured intracranial aneurysm by computed tomographic angiography. PMID- 9728256 TI - Growing skull fractures: a clinical study of 41 patients. PMID- 9728257 TI - [Anesthesiologic quality assurance]. PMID- 9728258 TI - [Heparin induced thrombocytopenia]. AB - Thrombocytopenia is a known adverse reaction occurring in some of the patients receiving heparin. Two types of heparin-induced thrombocytopenia (HIT) have been described. HIT type I is mild thrombocytopenia probably caused by a direct proaggregating effect of heparin and occurs during the first few days of heparin treatment. No specific treatment is necessary. HIT type II is a severe thrombocytopenia mediated by an immunologic mechanism where antibodies against heparin/platelet factor 4 (PF4) complexes play a major role. Thrombocytopenia usually commences 4-14 days after the onset of heparin administration. The incidence of HIT type II is below 3% and even lower when low-molecular weight heparin is used. The possible occurrence of life-threatening thrombembolic events may complicate the course of HIT type II. Diagnosis of HIT type II by clinical features alone is often difficult. A few laboratory tests are pertinent for diagnosing HIT type II including the 14C-serotonin assay, the heparin-induced platelet activation test and the heparin/PF4 ELISA. Immediate cessation of heparin administration is essential in the treatment of patients with HIT type II, if need be even without waiting for the result of the antibody search test. Several alternatives of anticoagulation for patients with HIT type II have been investigated in the past. Danaparoid-sodium as well as recombinant hirudin have shown promising results when used for this purpose. PMID- 9728259 TI - [Four year's experience with quality assurance in anesthesiology in Hamburg]. AB - PURPOSE: Since 1992 421,851 anaesthesias were documented by 39 institutes with a standard dataset issued by the German Society of Anaesthesiology and Intensive Care (DGAI). The project was run by the Association for Quality Assurance (EQS) in Hamburg. Some results of the evaluation of this datapool are presented questioning the feasibility of the project to support improvement processes within the participating institutions and which adjustments should be done considering these experiences. METHODS: Data from machine-readable protocols and documentation software representing all anaesthesia cases were recorded since 1992 in a pilot project and since 1994 compulsory with a standard dataset issued by the DGAI. Comparing statistics of these data was produced at the EQS project office. In the steering committee and in meetings of the project participants the contents, policy and results of the project office. In the steering committee and in meetings of the project were critically analysed and adjustments initiated whenever necessary. Validity of data and feasibility of the method used was also questioned on the background of comparable studies. RESULTS: With an incidence of 14.1% of all anaesthesias with special occurrences (AVB) the results are in the same range if compared with most other studies. However, documentation of data is not complete. This is assumed to be due to the large size of the dataset with 112 items and the additional workload associated with it. Nevertheless the participants mostly consider the project to be a very useful support for internal improvement projects. CONCLUSION: The project method used so far is not mature yet. The data set must be streamlined and rendered more concise, quality indicators should be defined and tested, and the availability of statistically proven limits of tolerance should be the immediate aims in the further development of the project. PMID- 9728260 TI - [Concentration-dependent changes in the latency and amplitude of somatosensory evoked potentials by desflurane, isoflurane and sevoflurane]. AB - PURPOSE: Comparison of the influence of desflurane, isoflurane, and sevoflurane on the parameters of cortical somatosensory evoked potentials (SEP). METHODS: A total of 41 patients were randomly allocated to either the isoflurane, desflurane or sevoflurane group. Following induction with propofol and intubation, concentration of the volatile anaesthetic was kept constant at 1.3, 1.0, and 0.7 MAC for 15 minutes each in randomised sequences. No opioids or N2O were used. Cortical somatosensory evoked potentials were recorded following median nerve stimulation at the wrist with 1.5 times motor threshold current. SEP were evaluated for latencies of peak N20 and P25 as well as peak-to-peak amplitude N20P25. Measurements at the end of the 15 minute equilibration intervals were compared by analysis of variance for repeated measurements. Latencies and the logarithm of the amplitudes were assumed to be normally distributed. RESULTS: SEP could be recorded in all patients and at all concentrations. Latency of cortical SEP increased with anaesthetic concentration in a linear manner. No differences in latency increase were found between the three anaesthetics (ANOVA). In contrast, the decrease in amplitude with increasing anaesthetic concentration was non-linear. It was large from control to 0.7 MAC, but small in the range between 0.7 and 1.3 MAC. Amplitude reduction was larger with isoflurane than with sevoflurane or desflurane. CONCLUSION: 1) Sevoflurane and desflurane are better suited for anaesthetic management during intraoperative electrophysiological monitoring than isoflurane, because SEP amplitudes are better preserved. 2) SEP amplitude is less altered by changing anaesthetic concentrations in the concentration range from 0.7 to 1.3 MAC than SEP latency. PMID- 9728261 TI - [Usefulness and diagnostic value of evoked potentials in patients with neurologic diseases in postoperative intensive care]. AB - PURPOSE: To analyse the diagnostic value of evoked potentials, 176 examinations in 71 sedated, ventilated ICU-patients who could only be examined neurologically on a very limited scale, were registered. We focussed on the evoked potentials, the results having prognostic relevance and being useful in the anatomical localisation of the pathological process. The Glasgow coma scale, neuroradiological findings and the data of the outcome status after hospital or rehabilitation discharge were therefore obtained. RESULTS: We could show in distinct cases how evoked potentials could make a contribution to localise a pathogenic process. Failures in peripheral nerves, brachial plexus, myelin, brainstem and cerebrum were detected, respectively excluded. In the vast majority of cases, suspected, symptoms were very precisely predicted. This was especially evident in patients suffering from head injury, hypoxia and spinal cord injury. We found that a good outcome can be expected even with high intracranial pressures if the repeatedly registered central conduction time stays normal. CONCLUSION: We conclude that non-invasive evoked potentials do enrich the bedside diagnostic pattern in sedated intensive-care unit patients. While neuroradiological methods only allow statements on morphological changing, evoked potentials demonstrate the functional status of the peripheral and central nervous system. This method is easy to learn and the cost involved is justified both financially and from the viewpoint of personnel expenditure. PMID- 9728262 TI - [The multiple trauma patient. I]. PMID- 9728263 TI - [Thromboysis in the pre-hospital phase: theoretical advantage or real benefit for patient outcome?: Pro]. PMID- 9728264 TI - [Thromboysis in the pre-hospital phase: theoretical advantage or real benefit for patient outcome?: Contra]. PMID- 9728265 TI - [Insulin as an anabolic: hypoglycemia in the bodybuilding world]. AB - Excessive body building may be dangerous. To promote athletic performance and to improve physical appearance many of the body builders abuse anabolic-androgenic steroids and other drugs. The abuse of insulin as an anabolic medication in this athletic community was followed by a case of severe hypoglycaemia in a body builder. A 30-year old male presented with cerebral symptoms of hypoglycaemia. Directly before an international competition he tried to stimulate muscle growth by using the hypoglycaemic stimulus to the growth hormone. To achieve this he injected 70 IE of a short-acting insulin subcutaneously, resulting in severe hypoglycaemia. After the initial administration of intravenous glucose by the paramedics, he lost consciousness and showed signs of convulsions. After orotracheal intubation by an emergency physician, despite of ongoing infusion of glucose the blood glucose concentration remained low as measured in the out-of hospital setting. Finally administration of additional glucose and glucagon in the intensive care unit was able to stabilize the metabolic system. In any case of severe hypoglycaemia, repetitive measurements of blood glucose even in the prehospital setting should be performed to detect the hypoglycaemia especially if athletes are concerned. PMID- 9728266 TI - [Overlapping atelectasis after left side thoracotomy]. PMID- 9728267 TI - [Consent and patient education in anesthesia. E. Bierman AINS 32:427-452 (1997)]. PMID- 9728268 TI - An SEM and TEM study of the transition of the bronchus to the parabronchus in quail (Coturnix coturnix). AB - The main objective was to analyse the transition of the bronchus to the parabronchus in birds and to describe its specific structure in an integrated light microscopic, transmission electron microscopic (TEM) and scanning electron microscopic (SEM) study. Lung tissue from immature and mature quail was subjected to standard processing for paraffin light microscopy, TEM and SEM after intratracheal inflation with fixative. In transverse paraffin and Durcupan semithin sections, the partition incompletely closing the broncho-parabronchial transition has the appearance of a crest-like fold delineating the entrance to the underlying parabronchial vestibulum. The core of the entrance fold is composed of loose connective tissue with free cells, and has a well-developed blood supply and innervation. Voluminous groups of smooth muscle cells are interconnected with those of neighbouring entrance folds and the interatrial septa. On the parabronchial surface and partly on the bronchial surface the entrance fold is invested with simple cuboid epithelium consisting exclusively of granular cells with lamellar inclusions. On the bronchial surface, they pass into ciliated columnar pseudostratified epithelium. At the root of the parabronchially orientated surface, they continue into the mixed population of granular and squamous atrial cells of the parabronchus. Among the granular cells of the entrance fold, scattered epithelial neuroendocrine cells are consistently present. The three-dimensional visualization demonstrated the oval form of the entrance window with a circular field of non-ciliated cells delineating the entrance to the parabronchial labyrinthine system. The general structural pattern of the entrance fold, together with the complex system of interatrial trabecles of the parabronchi underline the multifactorial function of a complex system submitted to the skeletal, regulatory and host defense of the gas exchange tissue. PMID- 9728269 TI - In vitro-model for Toxoplasma gondii invasion into neuroepithelial cells. AB - In order to study the interactions of Toxoplasma gondii and neuroepithelial cells morphologically and biochemically we established an easy in vitro model, which simulates cellular contacts in congenital toxoplasmosis. Monolayer cultures of neuroepithelial cells from 13-14-day-old mouse embryos were prepared containing the typical ventricular cell types found in an embryonic brain, such as young neurons, macroglial and microglial cells. Ultrastructural investigations on cultures incubated for 1, 5 and 30 min or 1, 6, 12, 24 and 48 h with T. gondii indicated that all three cell types had been invaded by the parasites, multiplying in parasite vacuoles by means of endodyogeny. Microglial cells had already been penetrated by trophozoites within one minute and showed up to 3 or 5 parasite vacuoles per cell. Neurons and glial cells were invaded within 5 min and contained only one vacuole per host cell. All the parasite vacuoles were bounded by a membrane and bordered by the rough endoplasmic reticulum and mitochondria of the host cell after a few minutes. The vacuoles also contained some membranic tubuli. After 30 min some neuronal neurites were destroyed while the perikarya seemed to be unchanged. After 6 h the cytoplasm of the microglia lost more and more ribosomes and organelles. Neurons and glial cells showed no alterations. After 12h large areas of the vacuole membrane were folded up and lay curled up in the vacuoles. After 24 h incubation T. gondii had destroyed nearly all the microglial cells. The ultrastructure of neurons and glial cells now began to change in the same way as shown for microglial cells. The organelles and cellular membranes disintegrated and after 48 h incubation nearly all the cells in the neuroepithelial cell culture had fallen to pieces. For an identification of T. gondii in vitro by light microscopy or for the characterization of the cell surface we tried to label the parasites with 11 different FITC-stained lectins. None of the tested lectins bound to the parasites. We conclude that our in vitro model for invasion of T. gondii in neuroepithelial cells opens an opportunity for studying the interaction of these cells or the pharmacological effects on this interaction under defined conditions. PMID- 9728270 TI - Ultracytochemical demonstration of glycogen in cone, but not in rod, photoreceptor cells in the rat retina. AB - The presence of native glycogen in photoreceptor cells of the rat retina has not been identified in the literature. We have studied this ultracytochemically. After perfusion with glutaraldehyde fixative, the eyes were enucleated, and the retinal tissues, postfixed with OsO4, were embedded in epoxy resin. Some tissues were treated with saliva before postfixation. Ultrathin sections, stained by the periodic acid-thiocarbohydrazide-silver proteinate (PA-TCH-SP) method or with uranyl acetate and lead citrate, were examined by electron microscopy. On routinely stained sections, glycogen particles seemed to be absent in the cytoplasmic matrix of the photoreceptor cells because they were indistinguishable from the numerous ribosomes. This was due to a similarity in size and electron density. After PA-TCH-SP staining, fine electron-dense reaction products appeared on small cytoplasmic particles (but not on ribosomes) in the inner segments, perikarya and synaptic terminals of a subpopulation of photoreceptor cells. These particles, 15-25 nm in diameter, were identified as beta-particles of glycogen because of their susceptibility to enzyme digestion. The glycogen-rich photoreceptor cells were thought to be cone cells by reasons of their morphological features, such as synaptic terminals, nuclei and outer segments. These results suggest that the cone, but not the rod, photoreceptor cells in the rat contain abundant glycogen. PMID- 9728271 TI - Lipidosis induced in rat uteri by high doses of tamoxifen. AB - The anti-estrogenic drug tamoxifen is an amphiphilic cationic compound and might therefore be expected to interfere with intralysosomal catabolism of polar lipids as has been previously reported with several other amphiphilic cationic drugs. The purpose of this study was to investigate whether there is lipidosis induction in the uterus. High oral doses of tamoxifen (100 mg/kg) were administered to 9 adult rats for 6-14 weeks. Their uteri were examined by light and electron microscopy. Lipidosis-like alterations were seen in the glandular epithelia and in the myometrium. The luminal epithelium was most severely affected. The highest degree of intraepithelial change was already observed after a short-term treatment (6 weeks). The results support the previously proposed concept of a relationship between the amphiphilic cationic character of a compound and its ability to cause intralysosomal storage of polar lipids after a high dosage treatment of these drugs in animals. PMID- 9728272 TI - Functional activity of immune cells in female MS-patients. AB - Since the well documented immunological disturbances in patients suffering from multiple sclerosis are especially attributed to T-cells we felt it expedient to look also for the functional activity of lymphocytes, monocytes, granulocytes and natural killer cells in the peripheral blood of female MS-patients (n = 17) and healthy controls (n = 17). While MS-patients showed decreased levels of total lymphocytes, CD2(+)- and CD8(+)-cells the percentage of HLA-DR(+)-monocytes was increased, indicating a high activity level of these immune cells, whereas we could find no differences in the functional tests of monocytes, granulocytes and natural killer cells between MS-patients and healthy controls. The percentage of HLA-DR(+)-T-lymphocytes increased with the duration of the illness and support the stronger consideration of clinical staging. PMID- 9728273 TI - Three-dimensional angioarchitecture of tongue corrosion casts from normal young rats. AB - The three-dimensional architecture of the vascular network of the rat's lingual papillae has been studied employing the corrosive resin cast technique. The casts of the microvasculature of these types of papillae (the small conical filiform, true filiform, fungiform and vallate papillae) have been observed under scanning electron microscopy (SEM). The microvascular arrangements of the filiform papillae consist of well-defined simple or twisted capillary loops. The fungiform papillae have a cylindrical form with a central hole, and the capillary network gives shape to the whole papilla. Finally, the capillary bed of the oval-shaped vallate papilla with its characteristic network and small hairpin-like loops was also examined. PMID- 9728274 TI - [Noninvasive analysis of cartilage volume and cartilage thickness in the human elbow joint using MRI]. AB - The aim of the study was to non-invasively analyse the cartilage volume and thickness in the human elbow joint with magnetic resonance imaging. 12 fresh frozen specimens (ages 20 to 69 yrs.) were investigated using a 1.5 T magnet, and a water-excitation FLASH-3D sequence at a resolution of 1 x 0.25 x 0.25 mm3. After linear interpolation to 0.125 x 0.125 mm2 in the image plane, the cartilages were segmented interactively with a Snake algorithm. Following three dimensional reconstruction, the cartilage volumes were determined, and the mean and maximal cartilage thickness computed by Euclidean distance transformation. The total cartilage volume of the human elbow joint amounted to between 3.90 and 7.17 ml (mean 5.5 ml +/- 20%). The humerus occupied 49 to 60%, the radius 15 to 27% and the ulna 20 to 29% of the total volume. The mean cartilage thickness ranged from an average of 0.9 (proximal part of the ulna) to 1.4 mm (capitulum humeri), and the maximal thickness from 2.3 mm (proximal part of the ulna) to 2.9 mm (distal part of the ulna). The ulnar cartilage showed a more inhomogeneous distribution than that of the humerus and radius. The interindividual variability of the cartilage thickness was less than that of the volume. There was no significant relationship of the volume with age (r = 0.11) or body weight (r = 0.51). However, based on the joint size (r = 0.71-medio-lateral extension of the articular surfaces) about 50% of the variability of the total cartilage volume could be predicted. The technique presented is suitable for designing computer models to investigate the load transmission and functional adaptation of diarthrodial joints, and for diagnosing and monitoring joint disease. PMID- 9728275 TI - Synkinesis between facial nerve and oculomotor nerve. A case report. AB - We present a case history of a young man suffering from a facial-oculomotor synkinesis. The findings speak in favour of an acquired synkinesis due to trauma. Most probably the injury occurred in the midbrain, in the area of the vertical gaze control center and/or the area of the levator palpebrae motoneurons. A congenital synkinesis due to embryonic malformation seems to be unlikely, because at birth no restriction of the eye-ball motility was present. PMID- 9728276 TI - Branches of the intracavernous internal carotid artery and the blood supply of the intracavernous cranial nerves. AB - With the increasing frequency of surgical operations to the cavernous sinus greater knowledge of the microanatomy of the cavernous sinus has become necessary. The most frequently seen complications during cavernous sinus surgery involve impairment of cranial nerves. This can occur due to direct damage or ischemia. For these reasons, it is important to know the arterial supplies to the cranial nerves in the cavernous sinus and the anatomy of these branches as well. 15 formaline fixed adult cadavers were used in this study. Before the dissections, the internal carotid artery and vertebral artery were filled with coloured latex on both sides. In this report, the intracavernous branches of internal carotid artery (I.I.C.A.) were identified based on the principles of Nomina Anatomica (1989) and compared with others. In our study we found that the segment of the abducens nerve which lies in Dorello's channel was supplied by the meningeal branch; from the point at which it pierces the cerebellar tentorium, the trochlear nerve is supplied by the tentorial cerebellar artery; the posterior cerebellar artery supplies the proximal segment of the oculomotor nerve that proceeds to the oculomotor triangle. Except for these, all the cranial nerves that were located on the lateral wall of the sinus cavernosus are supplied by the tentorial marginal branch and the branches of the lateral trunk. PMID- 9728277 TI - An anatomical study of the tympanic branch of the glossopharyngeal nerve (nerve of Jacobson). AB - Our study was aimed to examine the anatomic relationships of the tympanic branch of the glossopharyngeal nerve (GPN), namely the Jacobson's nerve (JN). The JN is the first branch of the GPN after having passed the jugular foramen. It contributes to the tympanic plexus on the promontory. It transmits secretory innervation to the parotid gland. Its possible role in the regulation of the middle ear pressure has also been hypothesized in terms of animal studies. Using microdissection techniques and high-resolution computed tomography (HRCT) scanning, the anatomic relationships and course of the JN were examined in eight formalin-preserved cadavers (16 sides). A morphometric analysis related to the JN was also performed both in the 16 cadavers and 40 dry-skull specimens. The JN emerged from the inferior ganglion of the GPN in all specimens. The mean distance between the ganglion and the genu of the GPN was 11.3 mm. The inferior 2/3 of the tympanic canal (TC) followed a vertical course, and then it ran anteromedially with an angle of 160 degrees to 170 degrees. The mean length of the TC was 9.5 mm. The TC was well-defined in all axial HRCT scans. In 2 cases the JN was entirely encased in a bony canal in the middle ear. A double JN was observed in one case. This study gives an additional information regarding the anatomy of the JN. PMID- 9728278 TI - Morphology and development of the cervical part of the sympathetic trunk (pars cervicalis trunci sympathici) in the pig (Sus scrofa L.) during the prenatal period. AB - The aim of our experiment was to establish the morphological changes occurring in the cervical part of sympathetic trunk of the pig during its fetal development. The experiment was conducted on 38 swine fetuses, divided into three developmental groups. The investigated section in 6-11-week-old fetuses usually consisted of 3 ganglia. In the oldest age group only 2 ganglia appeared--the middle cervical ganglion was usually not observed. The statistical parameters referring to the cranial cervical ganglion (standard deviation, coefficient of variation) increase with the age of the fetus, suggesting that the greater part of the development occurs at the end of gestation. The investigated ganglion was negatively allometric in relation to body length during the whole of gestation. The most frequent arrangements of the cervicothoracic ganglia noted in the fetuses investigated were: [GCca + GTh1 + GTh2] on the left side of the body, and [GCca + (GTh1 + GTh2)] on the right. PMID- 9728279 TI - Extraorganic vascular system of adrenal glands in human fetuses. AB - The injection method was used to study the origin and variability of the blood vessels forming the extraorganic vascular system of the adrenal glands. Studies were carried out on 40 human fetuses of a crown-rump length between 113 and 280 mm (14 to 28 weeks of fetal age). It was proved that the arterial blood supply during the fetal period is extremely variable in both the origin and the number of adrenal arteries, as well as in the asymmetry of the blood supply between the left and right adrenal glands. The three main origins of the suprarenal arteries are from the inferior phrenic artery, the abdominal aorta and the renal artery. The inferior phrenic artery is the main one supplying the suprarenal glands during the fetal period. A characteristic feature of the extraorganic venous system in fetal adrenal glands is the constant presence of the adrenal vein, including number, orifice and the main tributaries. PMID- 9728280 TI - A quantitative study on the retial arteries in one-humped camels. AB - The brain arteries of camels were injected with latex and the rostral epidural rete mirabile, which is situated inside the cavernous sinus at the base of the brain, was studied both grossly and quantitatively. The rostral epidural rete mirabile is formed by the retial branches of the maxillary and external ophthalmic arteries and also the internal carotid artery. When the ratio of the blood supply to the rete was estimated from the diameter of each vessel, it was found that the branches of the maxillary artery carried 76.41%, the branches of the external ophthalmic artery carried 10.26% and the internal carotid artery carried 13.33%. PMID- 9728281 TI - Effects of retinoic acid on pedicle and first antler growth in a fallow buck (Dama dama L.). AB - Unilateral injection of 10 mg all-trans retinoic acid (in 1 ml of castor oil) into the early pedicle anlage of a male fallow deer resulted in accelerated growth of the cranial appendage, and altered pedicle and first antler shape on the treatment side, whereas pedicle and antler growth on the control side, injected with 1 ml of vehicle solution only, was normal. It is concluded that retinoic acid is able to alter pedicle and first antler morphogenesis, presumably by affecting the positional information of the periosteal cells covering the incipient pedicle. PMID- 9728282 TI - The effect of infant colic on maternal self-perceptions and mother-infant attachment. AB - Mothers of infants who developed colic were compared with mothers of non-colic infants on their perceptions of parenting self-efficacy, and separation anxiety as well as their attachment relationship with their infants. Colic was identified prospectively through telephone contacts with mothers. Questionnaires on self efficacy and separation anxiety were completed when infants were 5 months of age. At 18 months of age mothers and infants (colic and non-colic) participated in a laboratory situation to measure attachment. Results revealed that mothers of colic infants reported feeling less competent as mothers. In addition, while mothers of colicky infants tended to have more separation anxiety than mothers of non-colic infants, they felt that these separations did not have a negative effect on their child. Finally, no differences were revealed for attachment classifications between colic and non-colic infants at 18 months. PMID- 9728283 TI - Links between maternal care and persistent infant crying in the early months. AB - A community sample was screened to select three groups of infants and their mothers according to how much the babies cried at 6 weeks of age, the peak age for infant crying. The three groups--of moderate (n = 55), evening (n = 38) and persistent criers (n = 67) and their mothers--were assessed by diary, observation and questionnaire measures of mother and infant characteristics and interactions at 6 weeks and 5 months of infant age. At 6 weeks, mothers of persistent criers spent more time interacting with and physically stimulating their babies. Below optimum maternal sensitivity/affection was linked to moderately increased crying in the infants overall. However, most mothers of persistent criers showed optimum sensitivity and affection, while no significant links between maternal sensitivity/affection and infant crying were found in the persistent crying group. By 5 months, when infant crying declined, the range and size of differences between mothers of persistent criers and other mothers declined. Home observations and a standard play measure failed to show group differences in maternal sensitivity, affection and intrusiveness at this age. The findings show that persistent infant crying in the early months often occurs in spite of high quality maternal care, so that in most cases the crying is probably not due to inadequate parenting. The need to distinguish general community cases from those at social or medical risk is emphasized and the findings' implications for professionals are discussed. PMID- 9728284 TI - Relationships between infant crying, birth complications, and maternal variables. AB - The aim of the study is to identify factors influencing infants' crying behaviour (total crying duration, length of crying bouts, and frequency of crying). The searching for variables explaining inter-individual differences requires a sufficient stability of the cry parameters at least over short time intervals. Thus, a second aim is to assess the intra-individual stability of the crying behaviour in an age range from 3 to 12 months. Sixty-two mother-child dyads participated in the study. The results show substantial stabilities of the crying behaviour of infants between 3 and 12 months of age. This is related to the amount of crying as well as to the pattern of crying shown by the children over a 24 h interval. The typical cry pattern is characterized by peaks at 12.00 h and early in the evening, although there are large individual differences between the children with regard to cry durations at each hour of the day. Regarding the cry durations at 3 months of age, birth complications and the perceived emotional atmosphere in the mothers' family of origin are the major contributing factors. PMID- 9728285 TI - Persistent crying in early infancy: a non-trivial condition of risk for the developing mother-infant relationship. AB - Infants between 1 and 6 months of age (mean age 3.6 months) who were referred to the Munich Interdisciplinary Research and Intervention Programme because of persistent crying and their mothers were examined and compared with an age matched community-based control sample with no current cry problem. Three groups, referred extreme criers, referred moderate criers, and controls, were compared with regard to measures of psychosocial and organic risks, the mothers' perception of her own psychological state and infant temperament, the quality of mother-infant relationship, and intuitive parenting in mother-infant face-to-face interactions. In comparison with general-community samples of infants with persistent crying, the present clinical sample represents a biased group with particularly high levels of infant distress for long periods of time, with problems of sleep-wake organization, neuromotor immaturity, and difficult temperament. Moreover, extreme crying was associated with a cumulation of organic and psychosocial risks, including high rates of prenatal stress and anxiety, maternal psychopathology and partnership conflicts. Mothers in both referred groups scored similarly low on feelings of self-efficacy, and high on depression, anxiety, exhaustion, anger, adverse childhood memories, and marital distress. Mother-infant relationships were more often distressed or disturbed among referred dyads than among controls, and 40% as compared with 19% showed dysregulatory patterns of interactional failures in face-to-face contexts. The findings suggest that factors related to parental care did not cause persistent crying, but functioned as maintaining or exacerbating factors. Dynamic interactions between persistent crying, difficult temperament, and parenting factors which compromise maternal resources and intuitive parenting may put such families at long-term risk for both relationship and behaviour problems. PMID- 9728286 TI - Crying in the first year of life: good news in the midst of distress. AB - Excessive crying and colic in the first 3 months of life remain as mysterious and unsolved clinical problems. The mystery is contributed to by the relative lack of long-term follow-up studies. The findings from four new follow-up studies of infants with prior colic are analyzed in an attempt to derive a clearer picture of what 'life after colic' might be like for parents, infants, and their interactions. The bad news is that, for a subgroup of infants and parents, especially those with substantial additional risk factors, early excessive crying may not resolve, but evolve into a more generalized 'persistent mother-infant distress' syndrome. For some mothers of infants with colic, the risk of depressive symptoms or decreased self-efficacy may be increased. However, there appears to be good news for a substantial majority of infants with colic and for their parents. This includes a significant reduction over time in the amount of crying, intact parental and infant capacities to be responsive in interactive contexts, no significant maternal stress, and normal attachment relationships. PMID- 9728287 TI - In-situ polymerase chain reaction: foundation of the technology and today's options. AB - "In situ PCR" is the marriage of two established technologies in molecular genetics, the polymerase chain reaction (PCR) and in situ hybridization (ISH). It is based on the amplification within intact cells or tissue sections of specific gene sequences, or mRNA species, to levels detectable by ISH and/or immunohistochemistry. Methods to achieve in situ PCR, while sharing fundamental steps, have differed between different laboratories. On the basis of our own experience, in situ PCR appears to be best suited for the detection of DNA in single cell preparations, in which fixation and pre-treatments can be optimally controlled. Emphasis is placed on the requirement for appropriate and meaningful controls at the multiple steps involved. It is instructive to the view the emergence of this new technology in perspective. In situ PCR has not developed in isolation and is just one of several creative approaches that have been employed in recent years to study nucleic acids (DNA and RNA) intracellularly. Some approaches are more suitable for detection of mRNA, or viral RNA, while others are more easily applied to chromosomal DNA. Some further techniques, such as the isothermal self-sustained sequence replication (3SR), refined in-situ transcription (PRINS), or high sensitivity histochemical detection systems, will complement or even add to the potential of situ PCR. It is highly probable that tests will emerge, based on investigation of unique genetic markers, with important roles in specialized diagnostic laboratories for the evaluation of viral diseases, as well as hematological and other malignancies. PMID- 9728288 TI - High performance Nanogold-silver in situ hybridisation. AB - Conventional in situ hybridisation (ISH) usually requires the presence of at least 10-50 copies of the nucleic acid sequence in question per cell. In situ PCR has been proposed as an alternative method, which may yield single-copy sensitivity, but shows a relatively high rate of false-negative or even false positive reactions. Very recently, possible alternatives have been described, which can be performed in routine laboratories without the need for expensive equipment. Streptavidin-Nanogold-Silver ISH is an easy-to-perform assay, which can be applied to detect low copy numbers of nucleic acid sequences in paraffin sections and cytological preparations. Its combination with labelled tyramides (TSATM = tyramide signal amplification, also known as CARD = catalysed reporter deposition) can achieve single gene copy sensitivity in detecting DNA viruses and also shows very high sensitivity for RNA detection. Possible applications include the early recognition of viral infection, cancer-associated genes, genetic diseases, and also the specific detection of mRNA. PMID- 9728289 TI - A robust method for isotopic riboprobe in situ hybridisation to localise mRNAs in routine pathology specimens. AB - In situ Hybridisation (ISH) to detect mRNA is widely applicable to studies of human pathology and experimental models including transgenic mice, where it can provide crucial information as to where a gene is expressed, that is not available in other ways. ISH was used to establish that relatively high levels of expression of E-cadherin mRNA were associated with long-term survival in colorectal cancer, before suitable antisera became available. There is increasing awareness that ISH can be used to help select between candidate disease genes found at a linked locus, and that ISH can help validate therapeutic targets. For instance, when several closely related receptor genes are expressed in a tissue, it may be possible to determine which combinations are expressed together in a single cell type. These diverse applications demand a robust method that works on a variety of clinical and experimental materials, and is easily interpreted. To date, the ICRF in situ Hybridisation Service has hybridised over 30,000 sections of principally formalin-fixed, paraffin-embedded tissues, with a high success rate. The practicalities of our preferred method are discussed and key steps for quality control highlighted. PMID- 9728290 TI - In situ reverse transcriptase-polymerase chain reaction: an innovative tool for hepatitis C virus RNA detection and localisation, and for quantification of infected cells. AB - In situ reverse transcriptase-polymerase chain reaction is a novel and exciting technique which couples the extremely high sensitivity of DNA amplification with the advantages of in situ hybridisation, allowing the preservation of cell morphology and the localisation of the positive signal within intact cells. In this brief report, we analyse and discuss the application of such a technique to the study of hepatitis C virus (HCV) infection, the most common cause of chronic liver disease worldwide. Moreover, given the lymphotropism of this virus, we describe here our own approach to detect and localise HCV RNA within intact peripheral blood mononuclear cells and to quantify the relative number of infected cells. PMID- 9728291 TI - Flow cytometry in situ polymerase chain reaction as an innovative approach to HIV 1 proviral DNA detection. AB - In situ polymerase chain reaction represents an intriguing method to couple the sensitivity of PCR technology with the preservation of cell morphology. The target of this technique is the detection of specific DNA or retrotranscribed RNA inside the cell. In this brief report, we describe an in situ polymerase chain reaction technique revealed by flow cytometry in order to reveal the human immunodeficiency virus type 1 (HIV-1) proviral DNA in peripheral blood mononuclear cells. We also discuss the key steps and parameters of our assay in comparison with current opinion on in situ polymerase chain reaction. PMID- 9728293 TI - Six-year-old archival chromosome preparations are still good biological reagents for repeated primed in situ labelling (rPRINS). AB - Six-year-old chromosome preparations of Eulemur coronatus were used for in situ localization of highly repetitive DNA sequences. The application of the repeated primed in situ labelling (rPRINS) technique allowed the detection of positive signals whereas conventional fluorescence in situ hybridization gave negative results on the same archival preparations. This paper reveals the possibility of rescuing archival chromosome preparations for both clinical and comparative cytogenetics. Moreover, the chromosomal localization of the ECO-SAT 503 bp highly repetitive DNA sequences were found to localize in the pericentromeric region of every chromosome, with the exception of chromosome 9. PMID- 9728292 TI - Comparison of isotopic and non-isotopic labelling for in situ hybridisation of various mRNA targets with cRNA probes. AB - In situ hybridisation methods to localise messenger ribonucleic acid (mRNA) targets in tissue sections or cell preparations using riboprobes can be successful with either isotopic or non-isotopic labelling. Investigators often wish to decide which labelling method provides the maximum specificity, sensitivity and resolution, with minimum nonspecific background. In this study we compared isotopic (35S) and non-isotopic (digoxigenin) labelling, using a variety of probes and paraffin-embedded tissues. The targets were human beta-actin and von Willebrand Factor mRNAs in archival human tissues; and mRNAs for two closely related trefoil factor family (TFF) peptides, TFF2 and TFF3, in rat duodenum. Patterns of localisation with both isotopic and non-isotopic probes were broadly similar for each target. The 35S labelling provided good contrast and sensitive detection under darkfield illumination, but the cellular or subcellular resolution of the target was less precise than that obtained with the digoxigenin labelled probes in transmitted light. Digoxigenin labelling in individual cells was more clearly demonstrated, but occasionally the contrast of positive staining with background was poor. The sensitivity of each method appeared to be similar for these high-abundance targets, therefore the choice between isotopic and non isotopic labels is dependent upon the aim of the study and the cellular resolution required. PMID- 9728294 TI - Effectiveness of hip prostheses in primary total hip replacement: a critical review of evidence and an economic model. PMID- 9728295 TI - Bone marrow and peripheral blood stem cell transplantation for malignancy. PMID- 9728297 TI - Nurses intensify worldwide efforts to ensure HIV/AIDS care for all. PMID- 9728296 TI - Screening for speech and language delay: a systematic review of the literature. PMID- 9728298 TI - Nursing ethics: the challenge of new directions in healthcare policy. PMID- 9728299 TI - A cross-cultural tool to identify continuing education needs. AB - Cost-controlled health reforms have created the need for training nurses to provide evidence-based care in new contexts. Planing for nurses' continuing education must thus target the appropriate personnel and simultaneously consider local healthcare needs and training requirements. To plan such educational programmes, a training needs analysis instrument was developed in the UK and used to compare the training needs of nurses in Australia, the USA and the UK. Below, the findings of this survey. PMID- 9728300 TI - Preparing nurses to care for minority ethnic communities. AB - UK healthcare policy is increasingly emphasizing the need for healthcare services to be responsive to ethnic diversity. In recognizing the important role that nurses and midwives play in healthcare delivery, the English National Board for Nursing, Midwifery and Health Visiting--the statutory professional body that oversees nursing and midwifery education in England-commissioned a two-year research project to examine the extent to which nurses and midwives were prepared to work in a multi-ethnic society. The study found that not all practitioners were equipped to provide appropriate care to multi-ethnic populations. Below an overview of the study, which identifies some issues that may be of relevance to other countries. PMID- 9728301 TI - Use of Giger and Davidhizar's Transcultural Assessment Model by health professions. AB - Nursing science is increasingly gaining recognition. Witness the growing use by other disciplines of nursing's models and ideas. One example is the adoption of the Giger and Davidhizar Model of Transcultural Assessment by non-nursing disciplines to understand and address the needs of a pluralistic multicultural society. Below the authors describe a model to understand culturally diverse clients and show how health professions are using it to provide culturally competent care. PMID- 9728302 TI - Testing nursing theory cross-culturally. AB - For nursing science to be recognized globally, nursing theory must be tested in various cultural settings to prove its universality. Below, an overview of a study of cross-cultural nursing theory development in three German-speaking countries of Europe and its implications for clinical practice. PMID- 9728303 TI - The use of isolated patellar prostheses in Sweden 1977-1986. AB - During 1977 to 1986, 106 patients had isolated patellar prostheses, with both patellar and femoral components, inserted in 20 different hospitals in Sweden. At follow up, 7 years (range 3 to 14 years) after operation, 9 patients had died so that 97 patients with operations on 113 knees were included in this study. Thirty three (38 knees) had been operated on before or after a prosthesis had been inserted. At follow-up, 75% were satisfied with the operation; 83% had improved, 59% could walk without an aid and 44% had no, or only occasional, pain in the knee. These results encourage the use of an isolated patellar prosthesis in cases of advanced and disabling localised patellar arthrosis. There is no place for this operation in the treatment of chondromalacia. PMID- 9728304 TI - A comparison of conservative and delayed surgical treatment of anterior cruciate ligament ruptures. A matched pair analysis. AB - A series of 60 matched and paired patients with complete rupture of the anterior cruciate ligament (ACL) was studied; 30 were treated conservatively, even though operation was recommended, and 30 were operated on within 35 months (range 18 to 48 months) after the ACL rupture. The average age was 34 years in each group. They were assessed 39 months after arthroscopy or reconstruction. At follow up, no patient had flexion of less than 100 degrees, 13 of the reconstructed knees had an extension deficit, but in only one was this more than 10 degrees. Thirty six percent of the reconstructed and 14% of the conservatively treated patients graded their sports activity as unlimited, while 13% of the ACL reconstructions and 21% of those treated conservatively were severely limited. The Lysholm, Cincinnati and OAK scores were significantly better in the reconstructions. The anterior drawer sign was positive in 24% of the ACL reconstructions and in 81% of the conservatively treated patients; 19% had a positive pivot shift after reconstruction compared to 75% of those treated conservatively. Even though there was a considerable number of patients with a decreased range of motion after ACL reconstruction because of the slow regime of postoperative mobilisation used, the results of operation are significantly better than after conservative treatment even when ACL reconstruction was carried out late after injury. PMID- 9728305 TI - Lengthening osteotomy of the fibula for post-traumatic malunion. Indications, technique and results. AB - Relative shortening of the fibula may occur after any type of ankle fracture when the lateral malleolus is involved. Patients complain of pain and restriction of their daily and sporting activities. Clinically, there is valgus of the hind foot due to abduction and lateral rotation of the talus. The goal of treatment is to restore the initial length of the fibula by a horizontal or Z-osteotomy, which will also correct the malposition of the talus. This study shows that the operative reconstruction of a widened mortise is a relatively safe procedure, independent of the type of osteotomy used. Lengthening of the fibula is an important step in the treatment of the painful ankle when the fibula is short after trauma, even when degenerative changes of the joint are already present. PMID- 9728306 TI - Exposure of the surgeon to radiation during surgery. AB - Exposure to radiation over many years increases the incidence of cataracts and promotes the development of carcinoma of the thyroid gland. A prospective study of 24 operative procedures involving minimal invasive techniques and fluoroscopic guidance was undertaken in order to measure the radiation exposure to the primary surgeon. The study was conducted during 8 K-wire osteosyntheses in fractures of the distal radius, 8 closed interlocking intramedullary nailings in fractures of the femur and 8 internal fixator procedures, with or without posterior autogenic transpedicular bone grafting, in fractures of the lumbar spine. Radiation was monitored with the use of high sensitive thermoluminescent dosimeters. Fluoroscopy was necessary during the procedures, with exposure times ranging from 55 s to 12 min 35 s. The radiation dose received per procedure ranged from 0.6 259.3 microSv and was well within the dose limits set by German law. PMID- 9728307 TI - Treatment of intertrochanteric fracture with the Gamma AP locking nail or by a compression hip screw--a randomised prospective trial. AB - The Gamma AP (Asia-Pacific) locking nail (GAPN) is a modification of the standard Gamma locking nail made especially for use in Oriental patients. We made a randomised prospective comparison of the compression hip screw (CHS) and the Gamma AP locking nail for the internal fixation of 60 intertrochanteric fractures of the hip in elderly patients by comparing perioperative details and analysing the radiographic and clinical results. The operation time for the GAPN group was shorter than for the CHS group and the intraoperative blood loss was lower. The Gamma AP nail enabled earlier mobilisation. We found no significant difference in the time to union and the length of sliding of the lag screw between the two groups. The decrease in the neck shaft angle in the Gamma nail group was significantly smaller than in the CHS patients. There were no significant mechanical complications, such as fracture of the femoral shaft or failure of fixation in the Gamma nail group. On the basis of our observations we conclude that the Gamma AP locking nail is more efficient than the CHS in the treatment of intertrochanteric fractures in geriatric patients. PMID- 9728308 TI - Impaction grafting and acetabular reinforcement in revision hip replacement. AB - A substantial loss of bone stock is frequently encountered at revision of a hip replacement. A mix of autologous and homologous bone chips is a biological method of filling the cavities. Reinforcement implants can be used to anchor the new prosthesis and to impact the bone graft, protecting it during healing. The goals of this study were to evaluate the clinical and radiological results after revision of cups with aseptic loosening. Follow-up examination of 81 revisions in 78 patients at 6.5 years (range 3 to 9 years) showed that 93% of the patients were satisfied with their results. One patient underwent a further revision because of recurrent dislocation of the femoral head, and one had a superficial infection. All the grafts were fused at 3 months after the operation. The bone stock had increased in every case, but 6 of them show some degree of graft resorption. No implant showed impending signs of loosening. These results were encouraging. The reinforcement implants allow sufficient primary fixation and secondary stability can be achieved with the impaction grafting. Careful preoperative evaluation and assessment at operation is important to match bone defects with the grafts and selection of the prosthesis. PMID- 9728310 TI - Problems with the Mennen plate when used for femoral fractures associated with implants. A report of 5 patients. AB - The Mennen plate has been used for osteosynthesis of 5 femoral fractures, 4 of which were at the tip of a prosthesis stem. Union was achieved in 2 fractures, and loss of fixation of the plate in three. PMID- 9728309 TI - Revision of failed hemiarthroplasty for fractures at the hip. AB - The records of 56 patients in whom a hemiarthroplasty, carried out for a femoral neck fracture, had been revised to a total hip replacement, were reviewed. The mode of failure was femoral loosening in 21, acetabular erosion in 26 and both in 5. Loosening tended to occur earlier than acetabular erosion. The median time to the onset of symptoms was 12 months and to revision 33 months. There were 38 major operative or postoperative complications at revision in 27 of the patients (48%). PMID- 9728311 TI - Pharmacokinetics, uses, and limitations of vancomycin-loaded bone cement. AB - We have studied the mechanical and pharmacokinetic characteristics of an industrially-prepared bone cement containing 3 g of vancomycin per 60 g cement. A low viscosity cement was selected, to increase contact between the antibiotic and the infected surfaces. Resistance of compression (95 mPa) was well above the required standard (70 mPa) and similar to that of other cements with or without gentamicin. The concentrations in blood, urine and bone were measured in mg/l and mg/kg, and compared to the break point (BP) of susceptibility tests, which must be obtained to achieve control of infection. Diffusion tests were conducted in vitro (elution in saline from rods), and in 30 sheep femora implanted with the cement in vivo. In the animal study, bone levels during the first three months were three-fold higher than the BP (i.e., were > or = 12 mg/l) in 92% of specimens from all areas of bone studied and at all times since implantation; they exceeded five times the BP in 56% of specimens and were never lower than twice the BP. The mean level was four times the BP after six months and fell sharply during the next six months. A pharmacokinetic study in ten patients who had a primary total hip arthroplasty with vancomycin-loaded cement as prophylactic antibiotic therapy showed that blood levels were lower than 3 micrograms/ml, i.e., 30 times lower than the toxic threshold (90 micrograms/ml). Vancomycin was undetectable in urine after the tenth day. The levels in drainage fluids were five times the BP after 24 h and equal to it after four days. None of the ten patients treated prophylactically with vancomycin-loaded cement developed evidence of allergy, toxicity, intolerance or loosening during a two year period. No adverse events were recorded in 17 other patients treated with a vancomycin (2 g) plus gentamicin (0.8 g) loaded cement as adjuvant therapy for severe prosthetic infection. PMID- 9728312 TI - The Girdlestone pseudarthrosis in the treatment of infected hip replacements. AB - The results of 78 resections of the head and neck of the femur (Girdlestone pseudarthrosis) in patients with infected hip replacements were studied. The mean follow-up was 5 years. At the time of the resection, gram-positive organisms were found in 53% of the cases, gram-negative in 33%, and in 12% there were mixed flora. The Girdlestone pseudarthrosis controlled the infection in 86% and achieved satisfactory relief of pain in 83%. The mean shortening of the limb was 4.1 cm and every patient needed some type of external walking aid. We found no correlation between the type of organisms and the persistence of infection, nor between shortening and the functional results. The Girdlestone pseudarthrosis is an acceptable method of controlling infection and relieving pain after infection of a total hip replacement. PMID- 9728313 TI - Serum albumin and deep infection in femoral neck fractures. A study of 437 cases followed for one year. AB - Four hundred and thirty-seven patients with femoral neck fractures were studied to determine the value of serum albumin estimations on admission. Serum albumin is a good predictor of mortality, and patients with low levels should be given additional nutritional support. We found that the serum albumin level is not useful in predicting deep wound infection. The infection rate of 3% does not justify the use of antibiotic prophylaxis in general. PMID- 9728314 TI - The importance of combined clinical and sonographic examination of instability of the neonatal hip. AB - A clinically unstable hip in a new-born may be an early sign of congenital dysplasia. Unless followed and treated at a young age, it can progress to a degenerative hip joint disorder with considerable functional disability in adult life. For this reason, the early diagnosis of neonatal hip instability is crucial. We present our experience with 9199 neonates examined independently by clinical and ultrasonographic techniques. Instability was diagnosed in 0.8% of the hips. Only 47% of the unstable hips were diagnosed by the initial clinical examination, in the remainder the dysplasia was recognised only by sonography. Sonographic changes were also detected on re-examination in 6% of the unstable hips following the recognition of clinical instability. It is evident that combined clinical and ultrasonographic examination significantly improves the detection rate of dysplastic hips in new-borns. PMID- 9728315 TI - Synovial biopsy. A comparative study from Saudi Arabia and Malaysia. AB - A comparative study of synovitis in Saudi Arabia and Malaysia was made with a view to determining any geographic variation in the incidence and pattern of the arthritides. The diagnostic spectrum in both series included pyogenic arthritis, rheumatoid arthritis, brucellar and tubercular arthritis, gout, pigmented villonodular synovitis, synovial chondromatosis and acute rheumatic fever. Date palm thorn synovitis was observed only in the Saudi Arabian series. While brucellar and tuberculous arthritis were predominantly seen in Saudi Arabia, the incidence of rheumatoid arthritis, pigmented villonodular synovitis and acute suppurative arthritis was almost equal in both countries. PMID- 9728316 TI - Slipped capital femoral epiphysis. A report of 4 cases occurring in one family. AB - We describe slipped capital femoral epiphysis in 4 members of a black, obese family, who were all first-degree relatives. The aetiology of slipped capital femoral epiphysis is unknown, although it is thought to be multifactorial. Genetic predisposition and environmental factors have been associated with the condition. A familial incidence with at least two cases in the same family has been reported. In epidemiological studies, this incidence ranges from 3% to 35%. Our cases were investigated in an attempt to find a possible aetiological genetic factor. A genetic predisposition with an autosomal dominant pattern of transmission is suggested, although environmental variables must be considered as provocative factors. PMID- 9728317 TI - Antibiotic-impregnated xenografts in the treatment of chronic osteomyelitic cavities. Seven cases followed for 3 to 5 years. AB - We describe seven patients with chronic osteomyelitis which developed in 3 following operation and in 4 after trauma. The treatment consisted of removal of dead tissue and filling the resulting cavities with gentamicin-impregnated xenografts. No antibiotics were used postoperatively. Urine gentamicin levels were above 0.5 microgram/ml for 8 days. The patients were followed up for at least 3.5 years and neither clinical nor laboratory signs of infection were detected. These results lead us the conclusion that gentamicin-impregnated xenografts may have a place in the treatment of chronic osteomyelitis. PMID- 9728319 TI - Psychological management, prevention and treatment of phantom pain after amputations for tumours. AB - Amputation for tumours is rarely carried out nowadays and has few specific technical features, apart from the rare cases where ingenuity is required to gain a few centimetres in length of a stump. As far as possible, the decision for amputation should not be imposed; it is better that the patient himself should take the initiative. The prosthesis and its constraints should be described honestly to avoid subsequent disappointment. Prevention of a painful phantom limb must always be undertaken, and based on certain operative and perioperative precautions. The most important factors are treatment by psychotropic agents and the quality of the human relationships between patient and surgeon. PMID- 9728318 TI - Giant cell tumour of bone with pulmonary metastases. A report of three cases. AB - Three cases of giant-cell tumour of bone with pulmonary metastases are reported. New metastases appeared and the existing nodules continued to progress after treatment in two patients who were kept under observation and no further treatment was given. When patients with giant cell tumours of bone present with lung metastases, it should be presumed that they are probably from the known tumour. However; tissue should be taken to establish the diagnosis and to exclude other concurrent lesions. Pulmonary metastases have a good long term prognosis, and so should be kept under observation, avoiding radical treatment such as lobectomy, chemotherapy and radiotherapy. PMID- 9728320 TI - Thyroid status affects the rat cardiac beta-adrenoceptor system transiently and time-dependently. AB - 1. The aim of this study was to investigate the time-dependency of the influence of dysthyroid states on the beta-adrenoceptor system in rat heart left ventricle. Therefore, the influence of acute and chronic hyper- and hypothyroidism on beta adrenoceptor-induced left ventricular responses, beta-adrenoceptor density, cardiac noradrenaline tissue concentrations, Gs alpha-proteins, and basal and stimulated adenylate cyclase activities was determined. 2. Hyperthyroid rats were obtained by feeding with thyroxine (T4)-containing rat-chow for 1, 4 and 8 weeks. Hypothyroidism was induced by adding 0.05% propylthiouracil (PTU) to the drinking water. Rats of varying ages were used in order to compensate for the differences in the duration of the treatments. Rats were aged 3 and 5 months at the end of the experiments. 3. Thyroxine treatment for 4 and 8 weeks increased the cardiac sensitivity to isoprenaline, but maximal induced inotropic responses were decreased. Cardiac ventricular beta-adrenoceptor density was increased only in rats treated with T4 for 1 week. This transient effect of hyperthyroidism on cardiac beta-adrenoceptor density was not observed in older rats. The PTU treatment resulted in a stable decrease of cardiac beta-adrenoceptor density. 4. Left ventricular tissue noradrenaline concentrations were unaffected by hyperthyroidism, where a decrease was observed in hypothyroid rats. Density of Gs alpha proteins was increased in hearts from chronic hyperthyroid rats. 5. These results indicate that the increased sensitivity to beta-adrenoceptor-mediated stimulation in chronic hyperthyroidism cannot be attributed to changes in cardiac beta-adrenoceptor density, but is probably caused by an enhanced content of Gs alpha. Accordingly, in hyperthyroidism, the beta-adrenoceptor system is influenced time-dependently, whereas hypothyroidism affects the beta-adrenoceptor system independent of time. PMID- 9728321 TI - Stimulatory effect of octopamine on beta 3-adrenoceptors to lower the uptake of [14C]-deoxy-D-glucose into rat adipocytes in vitro. AB - 1. The effect of octopamine on beta 3-adrenoceptors has been studied in isolated adipocytes of Wistar rats using uptake of [14C]-deoxy-D-glucose as the indicator. 2. Octopamine (0.1-1 nmol 1-1) induced a concentration-dependent decrease of [14C]-deoxy-D-glucose uptake into the adipocytes and this inhibition was not influenced by haloperidol at concentrations sufficient to block dopaminergic receptors. 3. Pindolol and propranolol reversed this inhibition of octopamine in a concentration-dependent manner. The effect of octopamine was reduced in the presence of Rp-cyclic AMPS triethylamine, the membrane-permeable antagonist of cyclic AMP (cAMP), indicating the mediation of cAMP in this inhibition. 4. A direct effect of octopamine on beta 3-adrenoceptors was proved using the application of antibodies. In the presence of an antibody for beta 3 adrenoceptors, the actions of octopamine were concentration-dependently reduced in a manner similar to the decrease of BRL37344-induced inhibitions. 5. The same degree of diminished activities for octopamine as that of BRL37344, the well known specific agonist of beta 3-adrenoceptors, was also obtained in isoprenline desensitized adipocytes. Insulin-stimulated uptake of [14C]-deoxy-D-glucose into adipocytes was not modified by isoprenaline induced desensitization. 6. These results suggest that octopamine can activate beta 3-adrenoceptors to lower the glucose uptake through an increase of cAMP in rat white adipocytes. PMID- 9728322 TI - Nitric oxide as neuromodulator of sympathetic transmission in rat vas deferens. AB - 1. Electrical field stimulation (EFS) of muscle strips in vitro elicited a tetrodotoxin (TTX)-sensitive biphasic contractile response consisting of a phasic component followed by a tonic one. 2. The amplitude of both components of the response was impaired by N omega-nitro-L-arginine and potentiated by sodium nitroprusside. Cystamine caused a reduction in amplitude of both phasic and tonic components of the response to EFS. Neither N omega-nitro-L-arginine, sodium nitroprusside, nor cystamine induced changes in the resting muscle tone, or in the contractile response to exogenous agonists ATP and noradrenaline (NA). 3. The nitric oxide scavenger, 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide, induced a reduction in amplitude of both components of the response to EFS. 4. These results reveal a facilitatory prejunctional modulatory role for nitric oxide in sympathetic neurotransmission in rat vas deferens. Endogenous nitric oxide released in the extracellular space is presumed to potentiate neurotransmission by acting at prejunctional level via cGMP. PMID- 9728323 TI - Contribution of cAMP to the inhibitory effect of non-steroidal anti-inflammatory drugs in rat uterine smooth muscle. AB - 1. The effect of the non-steroidal anti-inflammatory drugs naproxen, mefenamic acid, phenylbutazone, piroxicam and tolmetin on the vanadate (0.3 mM)-induced tonic contraction, as well as the modifications of these effects by the G-protein inhibitor pertussis toxin, and the inhibitors of protein kinase A, Rp-cAMPS (Rp Adenosine 3',5'-cyclic monophosphothioate triethylamine salt) and protein kinase C, H-7 [1(5-isoquinolynilsulfonyl)-2-methyl-piperazine], have been assayed to study the possible nature of intracellular mediators contributing to the inhibitory effects of NSAIDs in rat uterine smooth muscle incubated in medium lacking calcium plus EDTA. The effect of phorbol 12,13-dibutyrate on vanadate contraction and its modification with H-7 has also been examined. 2. Naproxen (6 600 microM), mefenamic acid (6-300 microM), phenylbutazone (6-300 microM), piroxicam (6-600 microM) and tolmetin (6-600 microM) produced concentration dependent relaxation of vanadate-induced tonic contraction. The potency order, in accordance with their respective IC50 values was: phenylbutazone > or = mefenamic acid > or = naproxen > tolmetin > or = piroxicam. 3. The relaxant effects of naproxen, phenylbutazone, piroxicam and tolmetin were significantly antagonized with pertussis toxin (50 ng ml-1), Rp-cAMPS (100 microM) and H-7 (1 microM). However, the effect of mefenamic acid was unmodified by the three drugs. This suggests that the effect of mefenamic acid and other NSAIDs occur by different mechanisms. 4. Phorbol 12,13-dibutyrate relaxed the vanadate contraction but the maximal relaxation achieved (54.8 +/- 8.3%, n = 4) was lower than those induced with the NSAIDs. On the other hand, H-7 (1 microM) did not modify the relaxant effect of phorbol 12,13-dibutyrate. This suggests that H-7 behaves as a PKA, but not a PKC inhibitor, under the present experimental conditions. 5. The relaxation by naproxen, phenylbutazone, piroxicam and tolmetin is presumably produced by increasing cAMP because the effects of these are antagonized with Rp-cAMPS and H 7, and by pertussis-toxin-sensitive mechanisms. PMID- 9728324 TI - Differential regulation of mesenteric and femoral blood flow in the rat as revealed by computerized data acquisition and evaluation. AB - 1. A set-up for computerized acquisition and evaluation of haemodynamic data was constructed. Blood flow (BF) in the superior mesenteric and femoral artery of urethane-anaesthetized rats was measured with the ultrasonic transit time shift technique. The signals for arterial blood pressure and BF were fed into a personal computer via an analogue-digital converter. Mean arterial blood pressure, heart rate and vascular conductance (CV) were calculated on-line. For subsequent analysis of the data, algorithms were programmed to filter the data, and to determine average and peak values for each parameter. 2. Systemic hypertension induced by phenylephrine (3-300 nmol kg-1), angiotensin II (0.1-3.0 nmol kg-1) and arginine vasopressin (0.03-1.0 nmol kg-1) was accompanied by constriction of the mesenteric artery. In contrast, the femoral artery responded to phenylephrine with constriction, to angiotensin II with dilatation and to arginine vasopressin with dilation followed by constriction. The haemodynamic effects of endothelin-1 (0.03-3.0 nmol kg-1) were generally biphasic, the initial hypotension being associated with dilatation, and the delayed hypertension being accompanied by constriction of both the mesenteric and femoral arterial bed. 3. Terbutaline (3-1.0 nmol kg-1) and calcitonin gene-related peptide (0.03-1 nmol kg 1) caused systemic hypotension along with mesenteric and femoral vasodilatation. 4. Telmisartan (1 mg kg-1), an angiotensin AT1 receptor antagonist, dilated the mesenteric artery, but had no effect on femoral VC. In contrast, the alpha 1 adrenoceptor antagonist prazosin (0.1 mg kg-1), dilated the femoral artery without altering mesenteric VC. Similarly, the beta-adrenoceptor antagonist propranolol (1 mg kg-1) had no effect on mesenteric VC, but constricted the femoral arterial bed. 5. These data demonstrate that the haemodynamic effects of exogenously administered drugs can widely differ between the mesenteric and femoral arterial beds of urethane-anaesthetized rats. Furthermore, vascular tone of these two arterial beds in maintained by different vasoconstrictor systems. While the femoral artery is mainly under adrenergic control, the renin angiotensin axis is predominant in the mesenteric arterial bed. In addition, this study also demonstrates that computerized analysis enables quick and accurate estimation of haemodynamic drug effects, and is superior to 'by hand' evaluation of peak changes in the functional diameter of the vascular bed under study. PMID- 9728325 TI - Effect of K+ channel blocking drugs and nitric oxide synthase inhibition on the response to hypoxia in rat pulmonary artery rings. AB - 1. The aims of this study were to investigate the effects of potassium (K+) channel blockers and the nitric oxide (NO) synthase inhibitor, L-nitroarginine (L NOARG), on the response produced by acute hypoxia in rat intrapulmonary artery rings in vitro. 2. In rat phenylephrine-precontracted pulmonary artery rings, hypoxia (pO2 = 7 mmHg) induced a response which consisted of a rapidly developing initial contraction (phase 1), a transient relaxation (phase 2) and a slowly developing sustained contraction (phase 3) over 30 min. The NOS inhibitor, L NOARG (300 microM), attenuated phase 1 and 3, and amplified phase 2 of the response to hypoxia. The voltage-gated K+ channel blocker 4-aminopyridine (4-AP) (10 mM) also abolished phase 3 and magnified phase 2 of the response to hypoxia. 3. The hypoxic response was not modified by the calcium-activated K+ channel (KCa) blockers, tetraethylammonium (TEA) (20 mM) or charybdotoxin (50 or 200 nM), nor by the ATP-dependent K+ channel (KATP), blocker, glibenclamide (10 microM). 4. L-NOARG (300 microM) and 4-AP (10 mM) also abolished carbachol-induced endothelium-dependent NO-mediated relaxation. Relaxation produced by the NO releasing agent 3-morpholino sydnonimine (SIN-1) was reduced by 4-AP (10 mM) and TEA (20 mM). 5. The data suggest that NO production is reduced during severe hypoxia in rat intrapulmonary artery rings and that this underlies the sustained phase of the hypoxic contraction. The data also suggests that 4-AP-sensitive K+ channels play an important role in the release and or action of NO, and therefore, in the response to hypoxia. PMID- 9728326 TI - Uptake of [3H]-adrenaline by freshly isolated rat hepatocytes: putative involvement of P-glycoprotein. AB - 1. The liver has an important role in the elimination of circulating catecholamines. Adrenaline and noradrenaline are avidly taken up and metabolized by rat hepatocytes, but the nature of the mechanism(s) involved remains partially unknown. 2. The aim of this work was to further characterize the uptake of catecholamines by isolated rat hepatocytes. For that purpose, the effects of a series of chemically unrelated compounds, including substrates/inhibitors of P glycoprotein, on [3H]-adrenaline removal was investigated. 3. Freshly isolated rat hepatocytes were incubated in Krebs-Henseleit solution at 37 degrees C with 50 nM [3H]-adrenaline for 5 min. Removal of [3H]-adrenaline was calculated as the sum of [3H]-adrenaline present in cells, and its [3H]-metabolites present both in cells and in the incubation medium. Radioactivity was determined by liquid scintillation counting. 4. Verapamil, quinidine, 1-methyl-4-phenylpyridinium, cimetidine, tetraethylammonium, d-tubocurarine, taurocholate, daunomycin and vinblastine (100 microM), progesterone, bilirubin (200 microM), vecuronium (45 microM), and amiloride (1 mM) significantly reduced [3H]-adrenaline removal. On the other hand, cyclosporine A (100 microM) apparently had no effect. The O methylated metabolite of adrenaline, metanephrine (30 microM), produced a 40% reduction of [3H]-adrenaline removal. 5. Vinblastine and corticosterone produced concentration-dependent decreases of [3H]-adrenaline removal, with IC50 values of 23.3 and 116.0 microM, respectively. 6. In the presence of verapamil (100 microM), desipramine (1 microM) was devoid of significant effect on [3H] adrenaline removal. Corticosterone (40 microM) produced a further decrease (+/- 50%) on removal of the [3H]-amine. 7. Removal of [3H]-adrenaline by isolated cells did not show pH-dependence since an increase or a decrease in the pH of incubation medium (to 8.2 or 6.2, respectively) caused no alteration of that parameter. 8. In conclusion, [3H]-adrenaline is efficiently removed and subsequently metabolized by isolated rat hepatocytes. The results are compatible with the involvement of multiple mechanisms in the hepatic uptake of this amine including the type I and the type II hepatic transporters for organic cations, uptake2 and P-glycoprotein. PMID- 9728327 TI - Komrower Lecture. Galactosaemia today: the enigma and the challenge. PMID- 9728328 TI - Sphingolipids and cell signalling. AB - The sphingolipid storage disorders constitute a group of inherited metabolic disorders in which the structure of the stored sphingolipid and the corresponding genetic defect have been established. However, the pathological mechanism(s) behind the disorders has not been fully elucidated. Sphingolipids are known to be recognition molecules involved in intercell communication and altered expression might lead to dyscommunication. The impaired degradation and lysosomal accumulation of specific sphingolipids might influence the metabolism of other molecules and/or intracellular transport, which in turn might alter the distribution of these molecules. However, the progress of these diseases indicates that additional factors, besides the stored sphingolipid itself, might be involved. During the last decade, several sphingolipids have emerged as active participants in intracellular signalling processes such as growth control and apoptosis. Particular interest focused on the sphingolipid metabolites, ceramide and sphingosine, as potential mediators in intracellular events and an altered presence of these metabolites in sphingolipidoses cannot be ruled out. Some sphingolipids have been found to influence cytokine release and thereby might induce immunological processes, which are known to exist in at least one of the sphingolipidoses--Gaucher disease. These processes might already have a pathogenic effect during early development, before significant storage has occurred. PMID- 9728330 TI - From toxicological problem to therapeutic use: the discovery of the mode of action of 2-(2-nitro-4-trifluoromethylbenzoyl)-1,3-cyclohexanedione (NTBC), its toxicology and development as a drug. AB - NTBC is a triketone with herbicidal activity that has been shown to have a novel mode of action by inhibiting the enzyme 4-hydroxyphenylpyruvate dioxygenase in plants. Early studies on the toxicity of this compound found that rats treated with NTBC developed corneal lesions. Investigations aimed at understanding the mechanistic basis for the ocular toxicity discovered that the rats developed tyrosinaemia and excreted large amounts of 4-hydroxyphenylpyruvate and 4 hydroxyphenyllactate, owing to inhibition of the hepatic enzyme 4 hydroxyphenylpyruvate dioxygenase. The corneal lesions resemble those seen when rats are fed a diet supplemented with tyrosine, leading us to conclude that the ocular toxicity seen with NTBC is a consequence of a marked and sustained tyrosinaemia. Studies in collaboration with Professor Sven Lindstedt showed that NTBC was a potent inhibitor of purified human liver 4-hydroxyphenylpyruvate dioxygenase. This interaction lead to the concept of using NTBC to treat patients with tyrosinaemia type 1, to block or reduce the formation of toxic metabolites such as succinylacetoacetate in the liver. Zeneca Agrochemicals and Zeneca Pharmaceuticals made NTBC available for clinical use and, with the approval of the Swedish Medical Products Agency, a seriously ill child with an acute form of tyrosinaemia type 1 was successfully treated in February 1991. Subsequently, other children with this inborn error of metabolism in Sweden and other countries have been treated with NTBC. The drug is now available to those in need via Swedish Orphan AB. PMID- 9728329 TI - Holoprosencephaly: a paradigm for the complex genetics of brain development. AB - Holoprosencephaly (HPE) is the most common major developmental defect of the forebrain in humans. Clinical expression is variable, ranging from a small brain with a single cerebral ventricle and cyclopia to clinically unaffected carriers in familial HPE. Significant aetiological heterogeneity exists in HPE and includes both genetic and environmental causes. Recently, defects in the cell signalling pathway involving the Sonic Hedgehog (SHH) gene, as well as defects in the cholesterol biosynthesis, have been shown to cause HPE in humans. These discoveries and current genetic approaches serve as a paradigm for studying normal and abnormal brain morphogenesis. PMID- 9728331 TI - Tyrosinaemia type I and NTBC (2-(2-nitro-4-trifluoromethylbenzoyl)-1,3 cyclohexanedione). AB - In tyrosinaemia type I (McKusick 276700), fatal liver disease results either because of liver failure during infancy or early childhood or because of development of hepatocellular carcinoma during childhood or adolescence. This is caused by toxic metabolites which accumulate because of deficiency of fumarylacetoacetase, the last enzyme in the tyrosine catabolic pathway. NTBC is a potent inhibitor of 4-hydroxyphenylpyruvate dioxygenase and has been shown to efficiently prevent tyrosine degradation, and production of succinylacetone, in patients with tyrosinaemia. Since the first trial of NTBC treatment for tyrosinaemia type I in 1991, over 220 patients have been treated by the drug using a protocol which includes regular follow-up with reports of clinical and laboratory investigations to the study centre in Gothenburg, where additional analysis of critical variables is done on regularly collected samples. The course of the disease in patients with acute tyrosinaemia has changed dramatically. Only 10% of the patients have not clinically responded to NTBC treatment. In half of these patients, successful liver transplantation has been performed which has further reduced the mortality rate during infancy to 5%. The international NTBC study has now been going for 5 years and data have emerged that indicate a decreased risk for early development of hepatocellular carcinoma in patients who started treatment at an early age. There are now 101 patients aged 2-8 years who have started NTBC treatment before 2 years of age, and no cancer has developed after 2 years of age among these patients. However, there is no safe age with respect to occurrence of liver cancer, which has been recognized at diagnosis at 1 year of age in one patient and after a few months of treatment in an infant who was given NTBC at 5 months of age. PMID- 9728332 TI - Therapeutic trials in the murine model of hereditary tyrosinaemia type I: a progress report. AB - We have studied a knockout mouse with fumarylacetoacetate hydrolase (FAH) deficiency as a model of human hereditary tyrosinaemia type (I (HT1). These mice have a phenotype very similar to the human disease, which is characterized by acute hepatic failure, renal tubular disease and hepatocarcinoma. We have previously reported on the efficacy of 2-(2-nitro-4-trifluoromethylbenzyol)-1,3 cyclohexanedione (NTBC) in preventing acute liver disease in HT1 mice. Here we present a progress report on long-term follow up (> 1 year) of high-dose NTBC therapy in combination with tyrosine restriction. In vivo retroviral gene therapy was also effective in abolishing the acute liver failure of HT1. Retrovirally treated mice remained completely healthy and active for 12 months after retroviral gene transfer. However, hepatocarcinoma developed in 2/3 treated animals after 1 year. Southern blot analysis showed that the tumours did not arise from retrovirally transduced hepatocytes but from non-corrected FAH deficient cells. These results highlight the extreme danger for tumour formation in HT1 and indicate the need for improved gene therapy that leads to the elimination of endogenous FAH-deficient liver cells. PMID- 9728333 TI - Mouse models of genetic disease: new approaches, new paradigms. AB - The mouse mutant resource is a valuable tool for gene function studies in the post-genomics era. However, despite a seemingly large catalogue of mouse mutants, it is recognized that we have access to mutations at only a small fraction of the total number of mouse genes. There is a phenotype gap that needs to be narrowed by the implementation of large-scale, systematic mutagenesis programmes in the mouse. Both genotype-driven and phenotype-driven approaches can be employed to recover new mouse mutations. Genotype-driven approaches include large-scale genome-wide mutagenesis by gene trapping in embryonic stem cells. For genotype driven approaches, the initial focus is on the characterization of the mutational change to the genome. Identification of the mutated gene is relatively trivial, but the genotype-driven route provides little indication of the likely phenotypic outcome of the mutation. In contrast, phenotype-driven approaches employ mutagenesis procedures that emphasize the recovery of novel phenotypes without prior assumptions about the underlying gene or pathway that has been disrupted- although identifying the underlying gene may not be trivial. One phenotype-driven approach includes chemical mutagenesis using N-ethyl-N-nitrosourea (ENU). ENU mutagenesis programmes are increasingly being brought to bear on increasing the breadth and depth of the mouse mutant resource, and in so doing narrowing the phenotype gap. PMID- 9728334 TI - Animal models of lysosomal disease: an overview. AB - The relative rarity of human lysosomal disorders, extremely heterogeneous genetic background and ethical restrictions make well-controlled studies difficult with human patients. Genetically authentic animal models complement human patients with their ready availability, homogeneous genetic background and the relatively flexible experimental designs. Spontaneous animal models of human lysosomal disorders are rare, particularly among small laboratory animals. However, the homologous recombination and embryonic stem cell technology has so far enabled us to duplicate almost all known human sphingolipidoses, two mucopolysaccharidoses and aspartylgly-cosaminuria in mice and more disorders are expected in the near future. This technology also allows generation of mouse mutants that are not known or are highly unlikely to exist in humans, such as 'double-knockouts'. Studies of lysosomal disease have come to the half-way turning point of the marathon race from clincopathological descriptions, identification of affected compounds, enzymology, to the present gene-level inquiries. The animal models will play an important role in our long journey from nucleic acids back to biology. While the utility of these mouse models is obvious, species differences in the brain development and metabolic pathways must be always remembered if the ultimate goal of the study is application to human patients. After all, the mouse is not human. PMID- 9728335 TI - Glycosphingolipid degradation and animal models of GM2-gangliosidoses. AB - Glycosphingolipids form cell type-specific patterns on the surface of eukaryotic cells. Degradation of glycosphingolipids requires endocytic membrane flow of plasma membrane-derived glycosphingolipids into the lysosomes as the digesting organelles. The inherited deficiencies of lysosomal hydrolases and of sphingolipid activator proteins both give rise to sphingolipid storage diseases. Recent research has focused on the mechanisms leading to selective membrane degradation in the lysosomes and on the mechanism and physiological function of sphingolipid activator proteins. The GM2-degrading system is a paradigm for activator protein-dependent lysosomal degradation. Three polypeptide chains contribute to the in vivo degradation of ganglioside GM2: the alpha- and beta chains of the beta-hexosaminidases and the GM2 activator. Mouse models of Tay Sachs disease (alpha-chain deficiency), Sandhoff disease (beta-chain deficiency) and GM2 activator deficiency have been described. While the phenotypes of these variants of GM2-gangliosidoses are only slightly different in humans, the animal models show drastic differences in severity and course of the diseases. The reason for this is the specificity of sialidase, which is different between mouse and human. A double-knockout mouse lacking beta-hexosaminidases A, B and S shows a phenotype of mucopolysaccharidosis and gangliosidosis. A substrate deprivation approach to therapy is discussed with respect to animal models of the GM2 gangliosidoses. PMID- 9728336 TI - Metachromatic leukodystrophy: molecular genetics and an animal model. AB - Metachromatic leukodystrophy (MLD) is a lysosomal storage disorder caused by the deficiency of arylsulphatase A (ASA; EC 3.1.6.8). Deficiency of this enzyme causes intralysosomal storage of the sphingolipid cerebroside sulphate. This lipid is abundant in myelin and it may thus not be surprising that storage mainly affects oligodendrocytes. Patients suffer from a progressive demyelination causing various neurological symptoms. The disease is fatal and treatment is not available. The human ASA gene has been cloned and more than 40 mutations have been analysed that cause metachromatic leukodystrophy. Few of these alleles are frequent among patients, whereas most mutant alleles have only been found in single families. Since MLD has only been described in humans and no naturally occurring animal model has been described, ASA-deficient mice have been generated by homologous recombination. The ASA knockout mice are unable to degrade sulphatide and store the lipid intralysosomally. The pattern of lipid storage in neuronal and non-neuronal tissues resembles that described for patients. In the nervous system, lipid storage is found in oligodendrocytes, astrocytes and some neurons. Animals display an astrogliosis and a decreased average axonal diameter. Purkinje cells and Bergmann glia of the cerebellum are morphologically aberrant. Demyelination is seen in the acoustic ganglion and occurs between the ages of 6 and 12 months. The animals are deaf at this age and display various neuromotor abnormalities. However, compared to humans the mice have a surprisingly mild phenotype, since they have a normal life span and do not develop widespread demyelination. ASA-deficient mice have been transplanted with bone marrow, which was transduced with a retroviral vector expressing arylsulphatase A. The majority of transplanted animals display sustained expression of arylsulphatase A from the retroviral construct up to 5 months after transplantation. However, preliminary data suggest that this therapeutic approach does not reduce storage material. PMID- 9728337 TI - Murine mucopolysaccharidosis type VII: the impact of therapies on the clinical course and pathology in a murine model of lysosomal storage disease. AB - Murine mucopolysaccharidosis type VII (MPS VII) is a lysosomal storage disease caused by a recessively inherited deficiency of the lysosomal enzyme beta glucuronidase. Affected mice have clinical, biochemical and pathological findings similar to those seen in humans with MPS VII (Sly syndrome), including growth retardation, facial dysmorphism, deafness, behavioural deficits and widespread glycosaminoglycan storage in lysosomes in the viscera, skeleton and brain. This mouse model is a useful tool for the evaluation of the effectiveness and experimental therapies for the MPS disorders. Syngeneic bone marrow transplantation performed in newborn MPS VII animals--before clinical evidence of disease is pronounced--prolongs life, improves hearing and bone growth, and prevents lysosomal storage in many sites, but does not correct the central nervous system disease. Enzyme therapy with beta-glucuronidase from the first days of life does reduce lysosomal storage in the brain in murine MPS VII. The enzyme-replaced mice also have reduced visceral lysosomal storage, impressive normalization of their phenotype and an improved life span. The effectiveness of gene therapy for the treatment of lysosomal storage disease has also been tested using the MPS VII model. When transplanted into MPS VII mice, syngeneic haematopoietic stem cells or mouse skin fibroblasts infected with retrovirus expressing beta-glucuronidase decreased storage, but only in the liver and spleen. Injection of an adenovirus vector expressing beta-glucuronidase into the vitreous of the MPS VII mice reduced storage in the retinal pigment epithelium and corneal endothelium. Intravenous administration of the adenovirus vector transduced with the beta-glucuronidase gene reduced liver and spleen storage and, when instilled into the cerebral ventricles, this viral vector caused beta glucuronidase production in epithelial cells lining the ventricles. Recently, retroviral vector-corrected MPS VII fibroblasts secreting high levels of beta glucuronidase were engrafted directly into the brains of adult MPS VII mice with resultant reduction in storage in neurons and glia adjacent to the grafts. Future efforts aimed at prolonging expression of the beta-glucuronidase gene by viral vectors and more precisely directing the therapeutic effect to the skeleton and brain will be important in optimizing treatments for murine MPS VII and extending the results of such therapies to humans with MPS. PMID- 9728338 TI - Mutant CuZn superoxide dismutase in motor neuron disease. AB - CuZn superoxide dismutase (CuZn SOD) is one of several antioxidant enzymes that defend the cell against damage by oxygen free radicals. Mutations of the SOD1 gene encoding CuZn SOD are found in patients with familial amyotrophic lateral sclerosis (FALS), a progressive and fatal paralytic disease that is caused by the death of motor neurons in cortex, brainstem and spinal cord. The disease can be reproduced in transgenic mice by expression of mutant human CuZn SOD. Recent studies both in vitro and in vivo suggest that the effect of mutation is to enhance the generation of oxygen radicals by the mutant enzyme. Thus, mutation converts a protective, antioxidant enzyme into a destructive, prooxidant form that catalyses free radical damage to which motor neurons are selectively vulnerable. Recent studies of neuroprotective agents in the FALS model show that inhibition of oxidative mechanisms (copper chelation therapy, dietary antioxidants, and coexpression of bcl-2) delays disease onset but does not extend disease duration. In contrast, inhibition of glutamatergic or apoptotic mechanisms (riluzole, gabapentin, and coexpression of glutamatergic or apoptotic mechanisms (riluzole, gabapentin, and coexpression of an inhibitor of caspase-1) has no effect on disease onset but extends survival by increasing the duration of symptomatic disease. Thus, neuroprotective agents differentially target the processes underlying disease initiation and propagation. PMID- 9728339 TI - The role of manganese superoxide dismutase in health and disease. AB - Manganese superoxide dismutase (MnSOD) is the mitochondrial enzyme that disposes of superoxide generated by respiratory chain activity. Of all electrons passing down the mitochondrial respiratory chain, 1-2% are diverted to form superoxide; thus production of hydrogen peroxide occurs at a constant rate due to MnSOD activity. Mice lacking MnSOD develop cardiomyopathy and basal ganglia lesions, have no lipid peroxidation products, but show destruction of enzymes with 4Fe-4S centres. Patients with complex I (NADH-CoQ oxidoreductase) deficiency show variable hyperinduction of MnSOD that is at least partially dependent on the extent of disturbance of redox state. This in turn appears to result in production of excess hydroxyl radicals, which are damaging to proteins, lipids and DNA. An alternative method of protection from oxygen radicals is employed by complex I-deficient cell types that do not induce MnSOD in that they show induction of the bcl-2 protein. PMID- 9728341 TI - Compromised fatty acid oxidation in mitochondrial disorders. AB - Measurement of palmitate oxidation by the tritium release method in cultured fibroblasts or in lymphocytes detects patients with mitochondrial beta-oxidation disorders and in addition many patients with dysfunction of the respiratory chain. The clinical presentation and studies of metabolite levels in serum and urine are valuable in the differential diagnosis between these two groups of disorders. PMID- 9728340 TI - Menkes disease: underlying genetic defect and new diagnostic possibilities. AB - Cloning of the gene defective in the X-linked neurodegenerative disorder Menkes disease led to a cascade of new findings. Besides giving a better understanding of the intracellular copper homeostasis, these findings had important consequences from a clinical point of view. Today the underlying genetic defect has been described in several patients affected by one of the three hereditary disorders of copper metabolism: Menkes disease, occipital horn syndrome and wilson disease. In this review we discuss mainly Menkes disease and the impact of the recent findings on the diagnosis of this disorder. PMID- 9728342 TI - Costs of F-18-FDG PET: experience with a satellite concept. AB - AIM: One of the main arguments against the acceptance and introduction of FDG PET as a regular benefit in the statutory Medical Insurance system in Germany are its (alleged) high costs. The aim of the present study has been to determine empirically over a period of twelve months the costs of FDG PET, making use of a satellite concept. METHODS: Documentation on performance and consumption data has been assembled for purposes of cost analysis. After analysis these data provided the basis for an assessment of the costs. In view of the high proportion of fixed overheads and strong fluctuations in consumption values, it was not possible to allocate reliably in an individual FDG PET investigation the various types of costs to individual causes. A monthly cost assessment procedure was therefore adopted. For this purpose data were recorded in the department for Nuclear Medicine, while the statistical assessment of the consumption data and analysis of the costs were undertaken in the Institute of Industrial and Hospital Management. RESULTS: Over the twelve month observation period (November 1995 to October 1996, 177 working days) 433 patients were included in the study: 244 with brain diseases 37 cardiac patients, 130 whole body studies and 22 combined studies (brain and whole body scans). The non volume based costs, maintenance charges and depreciation accounted for the highest proportion of the total costs (48%). The FDG-related costs accounted for 41% of the total costs, while the personnel costs amounted to 11% and the film costs to only 0.45%. The total costs incurred amounted to DM 1,205,050. CONCLUSION: These data represent a first empirical cost analysis for FDG PET based on a satellite concept and form a starting point for a cost/benefit analysis of this procedure. PMID- 9728343 TI - [Attenuation corrected myocardial PET after application of 18FDG: effect on the determination of regional myocardial uptake values]. AB - AIM: Post injection transmission measurement (PIT) can be performed using rotating 68Ge/68Ga linesources. This study estimates attenuation coefficients, count densities and relative regional uptake values of PIT corrected cardiac PET (E-PIT) compared to routinely pre-injection transmission measurement (RT). METHODS: A thorax-phantom with homogeneously filled myocardium or with simulated defects and six patients with advanced coronary artery disease were studied using ECAT Exact tomograph (Siemens CTI) equipped with three rotating linesources. Transmission was performed twice (PIT, RT), attenuation coefficients and emission data were analysed, the latter without attenuation correction (E-UK), corrected with PIT (E-PIT) and with RT (E-RT) (count density, standard and relative uptake values). RESULTS: Both in phantom and patient studies attenuation coefficients differed significantly between PIT and RT. Comparing E-PIT and E-RT, regional uptake values were different only in phantom simulation with myocardial radioactivity concentrations higher than 10 kBq x ml-1. The image contrast between defects and remaining myocardium in the phantom studies or the standard and relative uptake values in patient studies did not vary significantly. CONCLUSION: Under clinical conditions a post injection transmission measurement does not influence the accuracy of regional myocardial uptake values relevantly. PMID- 9728344 TI - [Investigation of the signal-to-noise ratio on a state-of-the-art PET system: measurements with the EEC whole-body phantom]. AB - AIM: The spatial resolution of PET scanners can be improved by using smaller detector elements. This approach, however, results in poorer counting statistics of the reconstructed images. Therefore, the aim of this study was to investigate the influence of different acquisition parameters on the signal-to-noise ratio (SNR) and thus to optimize PET image quality. METHODS: The experiments were performed with the latest-generation whole-body PET system (ECAT Exact HR+, Siemens/CTI) using the standard 2D and 3D data acquisition parameters recommended by the manufacturer. The EEC whole-body phantom with different inserts was used to simulate patient examinations of the thorax. Emission and transmission scans were acquired with varying numbers of events and at different settings of the lower level energy discriminator. The influence of the number of counts on the SNR was parameterized using a simple model function. RESULTS: For count rates frequently encountered in clinical PET studies, the emission scan has a stronger influence on the SNR in the reconstructed image than the transmission scan. The SNR can be improved by using a higher setting of the lower energy level provided that the total number of counts is kept constant. Based on the established model function, the relative duration of the emission scan with respect to the total acquisition time was optimized, yielding a value of about 75% for both the 2D and 3D mode. CONCLUSION: The presented phenomenological approach can readily be employed to optimize the SNR and thus the quality of PET images acquired at different scanners or with different examination protocols. PMID- 9728345 TI - [Diagnosis of sentinel lymph node in malignant melanoma: preoperative lymphoscintigraphy and intraoperative gamma probe guidance]. AB - AIM: The sentinel lymph node (SLN) has shown to reflect the histologic features of the remainder of the lymphatic basin in patients with melanoma and is of considerable prognostic relevance. Aim of the study was to localize the SLN pre and intraoperatively by means of lymphoscintigraphy and gamma probe guidance. METHODS: 38 patients with histologically proven malignant melanoma (tumor thickness > 0.75 mm) were preoperatively examined by injecting 40 MBq 99mTc Nanocoll intradermally around the lesion. The first lymph node identified was marked on the skin. Immediately after scintigraphy patients were referred to the operation room and intraoperatively mapped using a handheld gamma probe. Activity of the SLN and of the adjacent nodes was measured ex vivo. After excision of the SLN, the lymphatic basin was re-checked for radioactivity and activity of the SLN and of the adjacent nodes was re-measured after removal. RESULTS: The hottest reading was found in all patients in vivo and ex vivo in the preoperatively marked lymph node. Morphologically (macroscopically, ultrasound, CT) all nodes were unsuspicious. Histologically, in 8 patients metastatic involvement of the lymph node was found which led to a wide exploration of the lymphatic basin with consecutive lymph node excision in 7 patients. CONCLUSION: The findings suggest that combined preoperative lymphoscintigraphy and intraoperative mapping with a gamma probe is a powerful approach for exact localization of the SLN. Diagnostic detection of the SLN may have considerable impact for patient management, since extended lymph node dissection may be confined to patients presenting with positive SLN. PMID- 9728346 TI - [Positron emission tomography and bone scintigraphy of an osteoclast metastasis]. AB - We present the case of a 66-year-old-male patient who suffered from a bronchial carcinoma with a rib metastasis which was detected by PET staging with a relevant 18F-FDG-Uptake. The bone scintigram with 99mTc-DPD showed a defect in this area. The aggressive disease induced no osteoblastic response. PMID- 9728348 TI - [Strong lights and trends: German Society for Nuclear Medicine--1998 anniversary in Leibzig]. PMID- 9728347 TI - [Long-term follow-up and therapeutic control of a hepatic metastatic acinar cell carcinoma of the pancreas using FDG PET]. AB - A 33 years old woman presented with cramp-like abdominal pain. Ultrasound examination revealed multiple lesions in the liver of hyper- and hypoechoic echogenicity which in accordance to subsequently performed computed tomography and dynamic hepatobiliary scintigraphy were considered to be a focal nodular hyperplasia (FNH). A severe increase of the serum lipase concentration, suspected to be an acute pancreatitis, was treated conservatively and led to a short improvement of symptoms. Some months later, a severe progression of the pain symptoms occurred, along with a measurable expansion of the abdominal circumference and palpable tumors of the liver. The dynamic hepatobiliary imaging and the static liver scan showed a decreased perfusion and function of the nodes as well as a reduced RES activity, respectively. A subsequently performed Positron Emission Tomography (PET) with F-18-Fluorodeoxyglucose (FDG) showed a massively increased glucose metabolism of the liver tumors. The histologic result of several biopsies of the tumors revealed metastases of an acinus cell carcinoma of the pancreas. Under systemic and local chemotherapy, a temporary remission could be obtained that was clearly detectable in a second FDG-PET. Nevertheless, during the further course of the disease, a progression occurred being detectable in an additional control PET-study by an increase in tumor size as well as in tumor glucose metabolism. The patient died in liver coma 15 months after the histologic diagnosis was obtained. PMID- 9728349 TI - [Microsurgery]. PMID- 9728350 TI - [Replantation. Indications and organization]. AB - Although experience in replantation surgery increased during the past 30 years, there are still problems remaining. Refinement of microsurgical operations and improvement in rehabilitation technique have lead to superior functional results. Beside clear indications as thumb loss or replantation in children there are only few absolute contraindications. A retrospective analysis of more than 300 replantation showed that there are still problems concerning exhaustive supply of replantation centres. Because time is one of the many critical components of successful preplantation improvements are necessary in extra- and intrahospital organization of replantation surgery. PMID- 9728351 TI - [Free toe transplantation in reconstructive hand surgery]. AB - Since the invention of microsurgical techniques the free toe-to-hand transplantation is an established method for reconstruction of hand function. The indication for this method following thumb and finger amputations as well as congenital defects with aplasia of the fingers and the thumb is discussed in comparison to alternative operations. The operative technique as well as the functional and aesthetic results that can be expected are described by presentation of clinical cases. Since 1992 we performed 18 free toe-to-hand transplantations for the above mentioned indications. Depending on the primary situation the elementary hand function with grip and pinch could be restored in all our patients using this method. Restoring sensibility next to length, mobility and strength in one operation has to be considered as an advantage compared to alternative methods. PMID- 9728352 TI - [Nerve prosthesis. Current status]. AB - To date, every attempt to bridge neural defects with nerve prostheses has failed. Silicone tubes,polyglactides, caprolactones and other compounds containing polyglactin have not solved the basic problems of nerve regeneration. The profound rejection responses to foreign matter cause destruction of the ultrastructural units that are imperative for the regeneration of neurites. Positive results obtained with animal experiments have not withstood clinical testing since inferior regenerative performance can occur despite obstructed regenerative pathways. PMID- 9728353 TI - [Tumor-related arthrodesis. Reconstruction of the shoulder contour using a free TRAM (Transverse Rectus Abdominis Musculocutaneous) flap]. AB - Upper arm bone- and shoulder resection due to cancer have to be performed under oncologic criteria. Soft tissue however may not be sufficient to safely cover a following arthrodeses. To avoid perforation and reconstruct a normal shoulder contour a free myocutaneous TRAM-flap was employed and its vessels anastomosed to branches of A. and V. brachialis. Incorporating the facia and an island of M. rectus abdominis proved to safely cover the arthrodetic material and shape the bulk of the flap, allowing to reconstruct the desired contour. Good vascularisation postoperatively preserved tissue stability and volume and thus safely restored normal shoulder contour even in a long time review. PMID- 9728354 TI - [Transplantation of the M. gracilis in the reconstruction of hand function]. AB - Free muscle transplantation with motor innervation offers the possibility to add contractile elements to upper extremities with extensive loss of musculature due to direct trauma or untreated compartment syndrome (Volkmann's contracture). The functional cross section area and the mean resting fiber length determine the maximum power and the contracting amplitude of the donor muscle. Although considerably weaker than the finger flexors to be replaced, the gracilis muscle was the preferred donor muscle because of its neurovascular pedicle and the minimal donor site morbidity. In a series of 15 gracilis transplantations, all 13 muscles that survived regained function. Finger motion was dependent on the preoperative condition of tendons and joints. Even after complete loss of the flexor and extensor compartment a useful upper extremity could be restored, which was preferable to the only alternative-amputation. PMID- 9728355 TI - [Prefabricated flaps, a new reconstructive concept]. AB - Microsurgery has entered its third generation. Functional and aesthetic requirements have been steadily increased with the refinements of microsurgery. The new principle of prefabricated flaps are another step to match these requirements. These prefabricated flaps can either be created by expanding known flap donor sites or creating entirely new flaps by implanting AV-loops or vascularized fascia. Principles and indications of this new technique will be elucidated by clinical examples. PMID- 9728356 TI - [Microsurgery in lumbar disk operations. Possibilities, methods and results]. AB - The frequency of lumbar disc hemiation amounts to 5.1% with the male and 3.7% with the female population. Because of the often long-time pain-conditional impairment of the patients as well as the varied therapy-possibilities, the treatment of the ruptured lumbar intervertebral disc represents a special challenge. The indication to the operation for the lumbar disk-herniation results from the malfunctions of the nerve roots, the pains, as well as the temporal course of the symptoms. New and important developments have given the introduction of micro-surgical operation-techniques into the orthopedics. This development has led to it that many orthopedists and neurosurgeons the micro surgical operation-technique prefers. Important for the micro-surgical lumbar discectomy is the operation-microscope, a good preoperative diagnostics as well as a particular micro-surgical operation-instrument. Through the different enlargement-possibilities of the operation-microscope, all anatomical structures can increase and are done visibly for the surgeon as well as the assistant. Intraoperative injuries of the dura as well as the spinal-nerves are certainly avoided hereby. The micro-surgical discectomie requires no longer time like the conventional operation of the disk. The postoperative stay in the hospital as well as the time of the work-incompetence is reduced. Postoperative inflammations of the disk-area as well as renewed operations are rarer with the microchirurgischen technique. If an operation is necessary, so this should not be out-hesitated too long in order to avoid bed results. PMID- 9728358 TI - [Microsurgical transoral decompression in diseases of and injuries to the cranio cervical junction]. AB - Besides the microsurgical ventral decompression for treatment of cervical disc prolapses, combined with an intercorporal fusion using an autologous bone graft from the iliac crest and a plate osteosynthesis, the microsurgical, transoral approach to the craniocervical junction has proven to be an effective procedure for adequate indications. Even for surgical treatment of diseases and injuries of the craniocervical junction ventral, anterolateral, lateral and dorsal approaches are applicable alone or in combination. The special anatomic and functional conditions of this region, however, obviously require that the indicational criteria for the various approaches differ from those selected for the other cervical segments. The postoperative complication risk requires that particulary critical consideration be given to the question of isolated transoral interventions. The same holds true for the question as the necessity for additional ventral stabilisation in combined dorsoventral interventions. This report is about 20 patients who underwent transoral decompression, about the indications and the procedure typical problems. PMID- 9728359 TI - [Femur head preserving therapy, using vascular pedicled iliac bone graft, in segmental femoral head necrosis]. AB - Various therapeutic options have been proposed in the treatment of femoral head necrosis ranging from conservative management to total hip arthroplasty. Since microsurgical techniques are routinely used in orthopaedic surgery, the importance of revascularization has increased in recent years. Vascularized bone grafting as well as various osteotomies have been considered for the stages II and III according to Ficat and II, III and IV of the criteria of the ARCO system respectively, which also bases on MRI-findings. We investigated the results of 80 patients with avascular necrosis of the femoral head treated with a vascular pedicled iliac bone graft, perfused by the A. circumflexa ilium profunda, between 1988 and 1996. Mean follow-up was 5.6 years. The evaluation was based on the harris hip score, clinical and radiological examination as well as a subjective assessment using a VAS (visual analog scale). The clinical results according to the harris hip score were good or excellent in 86.1%. These results correlate with the subjective assessment of pain and of the hip joint function with an average of 7.9 points using the VAS (max. 10 points). Unchanged radiological appearance over the follow-up period was observed in 47.5% using the ARCO criterias. Reviewing the studies using vascularized grafts, about 50% of the patients with a stage II and III according to Ficat and II, III and IV of the ARCO respectively show an unchanged stage of the disease 5-6 years after the procedure. Therefore, transplantation of a vascular pedicled iliac bone graft possibly offers the chance to intervene causally in the course of the disease with only little alteration of the biomechanics of the hip joint. PMID- 9728360 TI - [Free vascularized bone transplantation in the extremities]. AB - There is a growing trend today which calls for reconstructing the loss of bigger bone parts in the area of extremities in a suitable manner. The microvascular bone transplantation for bridging bone defects is--admist other procedures--a distinct enrichment to preserve the extremities. This method of transplantation has the capability of surviving within a weakened transplant bed. Because of this capability one inevitably wants to know the criteria which determine the biological behaviour of the transplants. Furthermore, it is essential to known how this criteria can be best managed, considering the different indications and locations. The bone healing and bone hypertrophy of 81 patients who received vascularised bone transplantations have been examined with respect to different parameters. As the positive capacities of the vital transplants are almost exclusively dependent on the actual supply with blood, angiography have been undertaken during three months after surgery. 71 patients with a patent anastomosis after surgery have been evaluated. Differences in bone healing of the vascularised transplants have been observed in regard to the following parameter: -The tumor group showed a better rate of bone healing than those patients with trauma and congenital tibiapseudarthrosis.--The bone healing results of the group of younger patients were better than those of the group of older patients. Moreover the transplants without a history of infection were better compared with transplants with a history of infection. Clear differences of the fibula hypertrophy behaviour have been observed with respect to the following parameters: upper extremities < lower extremities, thigh < lower leg, longer transplants < shorter transplants, group of older patients (35-60 years) < group of younger patients (1-18 years), plates < screws. PMID- 9728361 TI - [Osteonecrosis of the femoral condyle]. PMID- 9728362 TI - Analysis of BAX expression in human tissues using the anti-BAX, 4F11 monoclonal antibody on paraffin sections. AB - BAX, a heterodimer partner of BCL-2, is an apoptosis inducer. We aimed to characterize the distribution of the BAX protein in normal adult human tissues using immunohistochemistry (IHC). The monoclonal antibody anti-BAX 4F11 was used on paraffin sections: immunodetection of BCL-2 was performed simultaneously on serial sections. The specificity of BAX IHC staining was verified by Western blot analysis. IHC positivity was correlated with the detection of a specific 21 kDa band on Western blots. BAX immunostaining was mainly cytosolic and occasionally on the nuclear membrane. Amounts of BAX protein were high in liver, renal tubules, endocrine islets of the pancreas, gastric glands, cardiac muscle, epididymis, lymph node germinal centers, and neurons; intermediate in the colon, stomach, bronchus. Fallopian tube, salivary gland, breast, thymus, spleen, and testis; low or undetectable in the other tissues. BAX IHC positivity correlated with apoptotic features in neurons and germinal center lymphocytes. There was no strict correlation between the IHC profiles of BAX and BCL-2 expression, although a reciprocal pattern of staining was observed in lymph node and colon. This report shows the usefulness the monoclonal antibody anti-BAX 4F11 on paraffin sections and demonstrates that the human BAX tissular distribution is close to, but not similar, to the profile observed in the mouse. The widespread BAX expression suggests that BAX alone is insufficient to trigger cell death in human tissues. BAX may either modulate the role of other regulators of apoptosis or carry out functions unrelated to apoptosis. PMID- 9728363 TI - The spatial distribution of pulmonary lesions in severe ARDS. An autopsy study of 35 cases. AB - The present study was undertaken in order to describe the local distribution and temporal course of pulmonary lesions in severe ARDS. We investigated a total of 35 patients (22 females), ranging in age from 2 to 51 years, who suffered from ARDS III and IV and were treated by extracorporeal CO2 removal and low frequency positive pressure ventilation (ECCO2-R). The extent of acute and chronic diffuse alveolar damage was assessed on histologic gross sections in the ventral, central and dorsal zone of the upper and lower lobes. The lesions showed a characteristic uniform distribution. Areas with chronic DAD were predominantly situated in the ventral portions of the upper lobes. Acute DAD predominated in the dorsal and basal areas of the lung. The extent of acute and chronic DAD was virtually independent of the duration of disease. Hemorrhage occurred at the interface zone between chronic and acute DAD and made up a significant volume portion of the lung tissue, ranging between 8% (lower lobes) and 42% (upper lobes). We conclude that the progression of acute DAD to chronic DAD is mainly determined by local factors (hydrodynamic and hydrostatic forces, intraalveolar pressure) that differ within the lung, whereas the duration of disease plays a minor role. Parenchymal hemorrhage occurs at the interface between areas of acute and chronic DAD and may therefore primarily be due to an increased susceptibility of the pulmonary parenchyma to mechanical stress. PMID- 9728364 TI - The neuropathology of organ transplantation: comparison and contrast in 500 patients. AB - The main objective of this report is to compare and contrast the type and frequency of neuropathological findings following liver, heart, lung, heart-lung, kidney and bone marrow transplantation and to provide an overview of the major systemic complications in patients that received allografts. This is a retrospective analysis of the complete autopsy records and clinical histories of 500 adults who underwent organ transplantation at the University of Pittsburgh Medical Center during the interval of March, 1981 through July, 1997. This study is based on the neuropathological and systemic findings among 225 liver, 101 heart, 40 lung, 28 heart-lung, 74 kidney and 32 bone marrow transplants. Clinico pathological correlations were made. All patients received base-line immunosuppressive therapy with one or more of the following drugs: cyclosporine, corticosteroids and azathioprine. Since August, 1989, the primary immunosuppressive agent is FK-506 (Tacrolimus). Some patients received antilymphocyte globulin (OKT3), when acute rejection was imminent. Light microscopic examination of tissue sections, stained with hematoxylin and eosin and in some cases with special stains were made. Ultrastructural evaluation were also performed in selected cases. All of the studies were carried out at the University of Pittsburgh Medical Center, Department of Pathology, Neuropathology Division. Cerebrovascular complications were the most frequent and were seen in 51% of the liver, 59% of the heart, 58% of the lung, 50% of the heart-lung, 49% of the kidney and 44% of the bone marrow allografts. Aspergillus sp. infection was the most common of all CNS infections followed by viral, bacterial and protozoal. Primary Central Nervous System Lymphoma (PCNSL) was seen in 2% of the liver, and 2% of the heart recipients. Post transplant lymphoproliferative disorder (PTLD) involving the brain was seen in 2% of the liver allografts, 3% of the heart, and 7% of the heart-lung recipients. PTLD systemically was seen in 6% of the liver, 7% of the heart, 5% of the lung, 11% of the heart-lung and 4% of the kidney allografts. Graft-versus-host disease was seen only in 41% of the bone marrow recipients. There was no statistically significant difference between the incidence of the total CNS complications among the different organ transplant groups (p value > 0.10), but there was a statistically significant difference in the systemic complications among the organ transplant groups (p value < 0.001). In conclusion that immunocompromised patients with impaired cellular and humoral immunity are at risk for the development of opportunistic infections and hematologic abnormalities. PTLD appears to be different in its pathogenesis from that of PCNSL which occurs anew in the brain of these patients. The neurological complications may be reduced by earlier recognition and better understanding of their etiopathogenesis. PMID- 9728365 TI - Etiological analysis of focal nodular hyperplasia of the liver, with emphasis on similar abnormal vasculatures to nodular regenerative hyperplasia and idiopathic portal hypertension. AB - Pathological studies were performed on 23 cases of focal nodular hyperplasia (FNH) under the hypothesis that FNH is a hyperplastic lesion caused by abnormal vasculatures of portal tracts within the nodule. For a comparison of the histological features of portal tracts, nodular regenerative hyperplasia (NRH), idiopathic portal hypertension (IPH), chronic hepatitis and so-called normal liver were used as control tissues. Extranodular areas of FNH nodules were also examined. Clinical data were briefly summarized. Most of the portal tracts within FNH nodules showed various abnormal findings, such as dilatation and/or stenosis of portal vein, muscular thickening of arterial wall with dilated or stenotic lumina, lymphocyte infiltration, and bile ductule proliferation. However, portal vein thrombi were not found. These findings were not thought to represent compensatory reaction to portal vein thrombosis. Similar abnormal features were also observed in extranodular areas of FNH although to a milder degree. These abnormal features resembled those of NRH and IPH. Moreover, the characteristic scar-like tissues within FNH nodules were proved to be abnormally large portal tracts including large feeding arteries, portal veins and bile ducts. It has been believed that septa and scar-like tissue within FNH nodules are not portal tracts and that arterial malformation independent of portal tracts are related to the development of FNH. In addition, venous structures within FNH modules have until now not been considered to be portal veins. However, this study revealed that severe anomaly of portal tracts including portal veins and hepatic arterial branches existed in FNH nodules. Moreover, portal tracts in extranodular areas were also abnormal. Clinically, only one patient had a history of oral contraceptives. Based on these findings, congenital anomaly of the portal tracts histologically resembling the abnormal portal tracts of NRH and IPH may be related to the pathogenesis of FNH. PMID- 9728367 TI - Malignant granular cell tumor: report of a case and review of the literature. AB - The histological, immunohistochemical and electron microscopic features of a rare malignant granular cell tumor (GCT) arising in the left radial nerve of a 54-year old man are reported. Despite a lack of local recurrence following extirpation, the tumor metastasized to the skull five years later. Light-microscopically, both primary and metastatic tumors consisted of markedly atypical or pleomorphic neoplastic cells with abundant cytoplasm containing diastase-resistant periodic acid Schiff reaction-positive granules. These tumor cells were arranged in a sheet-like pattern with mitotic figures including atypical ones, and were frequently immunopositive for proliferating cell nuclear antigen and c-MET, the c met proto-oncogene product. These findings reflect high-grade malignancy of the present tumor. In addition, the tumor cells were positive for S-100 protein and neuron-specific enolase. Ultrastructurally, a large number of intracytoplasmic granules featuring secondary lysosomes as well as long interdigitating cytoplasmic processes, intercellular intermediate junctions, discontinuous basal lamina-like structures, and stromal long-spacing collagen were observed. These findings indicated schwannian differentiation of the present tumor. In addition, based on a review of previously reported cases, the overall clinicopathological characteristics of malignant GCT were summarized. PMID- 9728366 TI - Electron microscopic evidence for the mechanism of blood-retinal barrier breakdown in diabetic rabbits: comparison with magnetic resonance imaging. AB - Diabetes leads to a breakdown of the blood-retinal barrier (BRB), which can be demonstrated in experimental models by immunocytochemistry and magnetic resonance imaging (MRI). The present study utilizes these methods to investigate the mechanism of BRB breakdown in diabetic rabbits, a model ideally suited to both procedures. Rabbits were treated with alloxan and examined 2 months, 1 year, and 1.5 years after the development of diabetes to assess BRB breakdown using MRI and immunocytochemical staining for endogenous albumin. Using MRI, an increased incidence of retinal vascular leakage is first evident at 1 year of diabetes. Electron microscopic immunolocalization of albumin suggests that BRB compromise is principally mediated by transendothelial transport of serum proteins in endocytic vesicle-like structures of approximately 0.4-1 micron diameter. Some additional retinal vascular leakage is occasionally demonstrated through the interendothelial cell tight junctions, but only when adjacent vascular endothelial cells show degenerative changes. The similarity of these findings to those previously reported for diabetic humans and rats supports the use of the diabetic rabbit as a model for studying BRB dysfunction. MRI and electron microscopic (EM) immunocytochemistry are complementary methods for evaluating BRB dysfunction. MRI can provide an overall picture of the entire eye without sacrificing the animal. EM immunocytochemistry can provide a more detailed picture of a limited area of interest to gain insight into the mechanisms of extravasation. Together, both methods provide a more complete understanding of BRB breakdown in diabetic rabbits. PMID- 9728368 TI - Anaplastic large cell lymphoma of the spleen. AB - A case is presented of CD30+ anaplastic large cell lymphoma of the spleen. The patient, a 61 year old woman with a history of chronic lymphocytic leukemia (CLL) was seen for the sudden development of splenomegaly with thrombocytopenia. A splenectomy was performed which showed massive replacement of the spleen by a population of large atypical lymphoid cells showing bizarre nuclear forms and multinucleated tumor cells reminiscent of Reed-Sternberg cells. Immunohistochemical studies showed strong membrane and dot-like paranuclear positivity in the majority of the atypical cells for CD30, with coexpression in many of the cells for CD15. Additionally, the cells also strongly reacted with CD3, UCHL-1, EMA and LCA. The present case illustrates an unusual variant of anaplastic (CD30+) large cell lymphoma sharing histologic and immunophenotypic features that overlap with those of Hodgkin's disease. The history in this patient of CLL with sudden development of splenomegaly raises the possibility of transformation of CLL into a high-grade lymphoma (Richter's syndrome). The possible pathogenetic implications of this phenomenon are discussed. PMID- 9728369 TI - Congenital hepatic fibrosis of heterotopic hepatic tissue. AB - We describe a unique case of a one-month-old infant with congenital hepatic fibrosis in the heterotopic liver within the adrenal gland. The main (orthotopic) liver was normal. Complex congenital heart disease and asplenia were associated. PMID- 9728370 TI - The teaching of pathology in undergraduate programs in medicine in Europe. PMID- 9728371 TI - Biomechanical testing of the lumbosacral spine. AB - OBJECTIVE: To assess various forms of anterior and posterior sacral fixation and to study the influence of anterior lumbosacral fixation and posterior pedicle fixation at L5 in conjunction with lumbosacral fixation. SUMMARY OF BACKGROUND DATA: Moments at the lumbosacral junction are high in the long constructs requiring lumbosacral fixation. The purpose of this study was to assess bending moments in flexion-extension and lateral bending and rotational forces at the lumbosacral junction involving a variety of long constructs to the lumbosacral junction. The incidence of pseudarthrosis in such constructs in the adult spine literature ranges from 7% to 40%. METHODS: An alignment jig was designed to display three-dimensional motion in the three orthogonal planes. Nine constructs of five specimens each were tested. These consisted of fixation at T12-L5-S1 (construct 1), T12-L5-S1 with anterior L5-S1 fixation and grafting (construct 2), T12-L5-S1, S2 with and without L5-S1 fixation and grafting anterior (constructs 3 and 4, respectfully), T12-S1, S2 with and without L5-S1 anterior grafting and fixation (constructs 5 and 6, respectfully), T12 Jackson intrasacral fixation with or without L5-S1 grafting anteriorly at the anterior fixation (constructs 7 and 8, respectfully), and T12-L5-S1, S2 fixation with anterior grafting only (construct 9). RESULTS: The use of anterior fixation statistically increased stiffness in extension. There was a trend toward increasing stiffness in constructs with anterior fixation (two anterior anterior-oblique L5-S1 screws) and in other loading modes as well. Failure to use L5 screw fixation significantly decreased torsional rigidity in long constructs without anterior fixation. CONCLUSIONS: In long constructs, particularly in scoliosis surgery requiring lumbosacral fixation, the addition of anterior fixation at L5-S1 is recommended. The addition of L5 fixation in addition to sacral fixation significantly decreases rotational stresses and is recommended as well. PMID- 9728372 TI - Helmet and shoulder pad removal from a player with suspected cervical spine injury. A cadaveric model. AB - STUDY DESIGN: Video fluoroscopy was used to evaluate the motion in an unstable spine during helmet and shoulder pad removal. OBJECTIVE: To observe the amount of motion that occurs during the removal of helmet and shoulder pads in an injured spine. SUMMARY OF BACKGROUND DATA: Removal of shoulder pads and helmet from a football player with suspected cervical spine injury can be particularly hazardous. How much flexion occurs at the unstable level during removal of equipment is unknown. METHODS: Six fresh cadavers were used in the study. In three, an unstable C1-C2 segment was created by transoral osteotomy of the base of C2. In the remaining three, instability was created at C5-C6 by a posterior release. Under fluoroscopic recording, the helmets were removed by first removing the chin strap, face mask, and ear pieces. With the neck stabilized, the helmet was carefully removed. The shoulder pads were carefully removed, with the head stabilized. Angulation, distraction, and space available for the cord were measured at C1-C2. Translation, angulation, distraction, and change in disc height were measured in the specimens with unstable C5-C6. RESULTS: In cadavers with C1-C2 instability, the mean change in angulation was 5.47 degrees, and space available for the cord was 3.91 mm. Shoulder pads were removed while the head was stabilized. The mean change in angulation at C1-C2 was less during removal of shoulder pads than during helmet removal at 2.9 degrees. Space available for the cord was 2.64 mm. Distraction was also greater during helmet removal (2.98 mm) than during shoulder pad removal (1.76 mm). In the unstable spine, the change in displacement in translation was greater during shoulder pad removal (3.87 mm), than during helmet removal (0.41 mm). Disc height change was similar. Distraction of the spinous processes was greater during helmet removal (3.68 mm) than during shoulder pad removal (1.37 mm). Angulation was similar in both maneuvers. CONCLUSIONS: Helmet and shoulder pad removal in the unstable cervical spine is a complex maneuver. In the unstable C1-C2 segment, helmet removal causes more angulation in flexion, more distraction, and more narrowing of the space available for the cord. In the lower cervical spine (C5-C6), helmet removal causes flexion of 9.32 degrees, and during shoulder pad removal the neck extends 8.95 degrees, a total of approximately 18 degrees. Disc height changes from 1.24 mm of distraction to 1.06 mm of compression during helmet removal and shoulder pad removal for a total 2.3-mm change. Translation, which correlates with the change in the space available for the cord, is greater at C5-C6 during shoulder pad removal. Because most of the cadavers had C5 anteriorly displaced on C6 to begin with, the extension force during shoulder pad removal caused a 3.87-mm change in reduction of C5 on C6. Because of the motion observed in the unstable spine, helmet and shoulder pad removal should be performed in a carefully monitored setting. They should be removed together by at least three, preferably four, trained people. PMID- 9728373 TI - Disc degeneration and cervical instability. Correlation of magnetic resonance imaging with radiography. AB - STUDY DESIGN: An imaging study was designed to evaluate disc degeneration and segmental instability in the cervical spine. OBJECTIVES: To compare the magnetic resonance imaging assessment of disc degeneration with the conventional plain radiographic evaluation of cervical segmental instability. SUMMARY OF BACKGROUND DATA: No studies have been conducted to investigate the association of disc degeneration with cervical instability. METHODS: Two hundred sixty consecutive patients with suspected cervical spine disorders were analyzed for horizontal and angular displacements on lateral flexion and extension radiographs and disc degeneration on T2-weighted magnetic resonance images of the cervical vertebrae. RESULTS: In all intervertebral levels, the grade of disc degeneration increased significantly (P < 0.01) with age. Cervical instability was identified in 151 segments (14.5%) and correlated with Grade 1 and Grade 2 degeneration in the intervertebral discs (P < 0.01). CONCLUSIONS: Cervical segmental instability may indicate early degeneration of intervertebral disc in the cervical vertebrae. PMID- 9728374 TI - Safe lateral-mass screw lengths in the Roy-Camille and Magerl techniques. An anatomic study. AB - STUDY DESIGN: Investigation of the mean safe lateral-mass screw lengths in the Roy-Camille and Magerl screw techniques in cadaveric cervical specimens. OBJECTIVES: To report the mean screw path length and to evaluate the relation of the screw trajectory to the nerve root in the Roy-Camille and Magerl screw techniques. SUMMARY OF BACKGROUND DATA: Potential injury to the cervical nerve root caused by too long a screw remains a major concern. Few studies regarding proper screw length and its relation to the adjacent nerve root are available. METHODS: Fourteen cervical spines were used for this study. Each lateral mass from C3 to C7 was drilled according to the techniques described by Roy-Camille (right side) and Magerl (left side). The cervical spines were harvested from the cadavers, and the anterior aspect of the lateral mass and spinal nerve were exposed. The screw path length between the dorsal and ventral cortices of the lateral mass were measured. An additional measurement was taken from the ventral aspect of the lateral mass to the nerve root along the screw path. RESULTS: The mean screw path length in the Roy-Camille technique decreased consistently from C3 (15.7 +/- 1.7 mm) to C7 (11.3 +/- 0.8 mm). The mean distance from the ventral cortex to the nerve root ranged from 1.2 to 2.3 mm, and the smallest value was at C7. The mean screw path length in the Magerl technique also decreased from cephelad to caudal, with a range of 15-16 mm at C3-C6 and a mean value of 13.8 mm at C7. CONCLUSIONS: A safe screw length is 14-15 mm in the Roy-Camille technique and 15-16 mm in the Magerl technique at C3-C6. A short screw may be used at C7 if desired. PMID- 9728375 TI - Dynamic motion study of the whole lumbar spine by videofluoroscopy. AB - STUDY DESIGN: Dynamic lumbar flexion-extension motion was assessed by videofluoroscopy. OBJECTIVES: To identify the motion patterns of the whole lumbar spine in normal subjects and in patients with low back pain or spondylolisthesis during actual movement. SUMMARY OF BACKGROUND DATA: Assessment of lumbar instability on terminal radiographs is controversial. Information regarding spinal kinematics during actual movement in vivo is scarce. METHODS: Fluoroscopic lumbar sagittal motion videos were recorded in volunteers (n = 13; mean age, 22.3) and in patients with chronic low back pain (n = 8; mean age, 43.5) and degenerative spondylolisthesis (n = 8; mean age, 63.1) while the subjects bent forward from a standing neutral position (eccentric motion) and then returned to the original position (concentric motion). The videos recorded approximately 8 seconds of motion and were converted to still images at 5 frames per second. Disc angles from the horizontal line were measured to estimate sagittal rotation of each segment. Disc degeneration was evaluated on T2-weighted midsagittal magnetic resonance image. RESULTS: In the volunteer group, six exhibited sequentially spreading motion, four exhibited simultaneous motion, and three showed an altered motion-spreading pattern in the eccentric phase. The first two patterns were considered normal. Six (67%) of the patients with chronic low back pain also showed normal patterns, but seven (88%) of the patients with degenerative spondylolisthesis showed disordered patterns. The order of motion in the concentric phase was also different among the three groups. Prolonged deflection of the slipped segment was observed more frequently in the patients with degenerative spondylolisthesis. Disc degeneration was not always associated with motion-spreading order and the motion patterns. CONCLUSION: Segmental instability influences the whole lumbar motion in patients with degenerative spondylolisthesis. The patients with chronic low back pain did not show a significant difference when compared with the volunteers. PMID- 9728376 TI - Compensatory spinopelvic balance over the hip axis and better reliability in measuring lordosis to the pelvic radius on standing lateral radiographs of adult volunteers and patients. AB - STUDY DESIGN: Sagittal alignments, including lumbar lordosis and spinopelvic balance (measured from C7, S1, and hip axis reference points for the relative positions of the spine and sacropelvis over the hips), were studied on standing 36-in. lateral radiographs of adult volunteers (control subjects) and patients who had specific spinal disorders. OBJECTIVES: To determine the most reliable methods for measuring lumbopelvic lordosis and to define significant spinopelvic compensations for sagittal balance. SUMMARY OF BACKGROUND DATA: Measurements for standing sagittal balance, obtained using a C7 plumb line, and segmental angulations of the spinal vertebrae, including lordosis to the sacrum, have been reported. Absolute values, even for normative data, have had wide variation and limited clinical usefulness. Correlations of sagittal balance with the reported spinopelvic angulations (spinal vertebral and sacropelvic angulations) have not been well defined. In addition, determinates of balance (spinal and pelvic) have not been studied for reliability, and compensatory mechanisms for maintenance of balance have not been carefully evaluated. Better recognition of the correlations and more reliable methods to measure lordosis and balance and the spinopelvic compensations for its maintenance may be beneficial in treating patients who have spinal disorders. METHODS: Measurements on standing 36-in. lateral radiographs were made for sagittal alignments in adult volunteers (n = 50) and in adult patients who had symptomatic degenerative lumbar disc disease (n = 50), low grade L5-S1 isthmic (lytic) spondylolisthesis (n = 30), and idiopathic or degenerative scoliosis (n = 30). All participants exhibited clinical compensation for balance. Data were analyzed for significant correlations within each group to determine compensatory correlations of spinopelvic balance with the other sagittal alignments. Intraobserver and interobserver reliability for the parameters evaluated were calculated. This included two methods for determining lordosis (S1 end-plate and pelvic radius techniques). RESULTS: Plumb line measurements for balance from the S1 and hip axis reference points, as defined, were similar in all four groups. However, the groups appeared to adjust for balance by using common and distinctive spinopelvic compensations that resulted in significantly and characteristically different angular alignments among the four groups. Lordosis and balance measurements were closely correlated, and the correlation was characterized by pelvic rotation and translation around the hip axis. The subjects with less lordosis typically stood with the C7 plumb line anterior to and at a longer distance from the sacral reference point. This was primarily because of posterior sacropelvic translation around the hip axis and not because the sagittal plumb line initially moved anteriorly away from the sacrum. This was true in all four groups and gave the appearance that the sacropelvis was less well balanced over the hips in the subjects with less lordosis. Even small differences in lordosis appeared to be associated with considerable adjustments in the other spinopelvic alignments. Therefore, it was important to determine that lordosis was lumbopelvic more reliably measured by the pelvic radius technique. CONCLUSIONS: Lower lumbar lordosis, by the pelvic radius technique, and compensatory sacropelvic translation around a hip axis, in addition to measurements from this axis to the C7 plumb line, were the primary determinates and most reliable radiographic assessments for sagittal balance. Understanding the common and characteristically different compensations that occur with balance in these patients who had specific spinal disorders may help to improve their care. PMID- 9728377 TI - Assessment of spinal musculature using surface electromyographic spectral color mapping. AB - STUDY DESIGN: A technique is described for analyzing electromyogram data from lumbar spinal muscles, with special reference to discrimination of people with back pain. The ability to discriminate was evaluated in 99 people (61 healthy and 38 with chronic or acute back pain), and the influence of load was assessed. OBJECTIVES: To evaluate methods of analysis of complex electromyogram data and to assess correlation of electromyogram information with clinical condition in people with and without back pain. SUMMARY OF BACKGROUND INFORMATION: In previous analyses of electromyogram data, only a small part of the data have been used. Spinal muscular decompensation has been postulated in chronic low back pain, but there has been no direct demonstration of this phenomenon. Objective measures are still elusive. METHODS: Lumbar spinal surface electromyograms were recorded during an isometric lifting task. The data were represented graphically as color coded plots of electromyogram frequency, time, and electromyogram amplitude. Spectral width at half-peak amplitude (spectral half width) was calculated from the digitized, summed data. Ninety-nine people were tested: 48 men (29 with no recent [in the past 2 years] history of back pain, 16 with chronic back pain, 3 with acute back pain) and 51 women (32 with no recent back pain and 19 with chronic back pain). RESULTS: Spectral color maps in people with chronic back pain were markedly different from those in healthy people. Spectral half width was greater in people with chronic back pain than in healthy people (P < 0.01). Blinded analysis of spectral color maps allowed discrimination of people with back pain from healthy people with a sensitivity of 76% and a specificity of 81%. People with a history of back pain had markedly variable half widths. CONCLUSIONS: A new method of analysis of electromyogram data from lumbar spinal muscles has allowed discrimination between people with back pain and healthy people. This provides direct evidence of a correlation between muscular electrical function, measured by electromyogram, and low back pain. This technique may have potential as an objective measurement tool. PMID- 9728378 TI - Can it be predicted which patients with chronic low back pain should be offered tertiary rehabilitation in a functional restoration program? A search for demographic, socioeconomic, and physical predictors. AB - STUDY DESIGN: A prospective clinical trial was conducted that involved six groups of patients with chronic low back pain selected from a large cohort (N = 816). OBJECTIVES: To correlate pretreatment baseline variables with outcome parameters after treatment in a functional restoration program or in control programs, to identify possible factors predictive of the need for functional restoration. SUMMARY OF BACKGROUND DATA: Since the functional restoration program was first described, research has focused on identifying patients who will or will not benefit from such a program. The value of previous studies is limited, however, because predictive factors from a control group were not "subtracted." METHODS: Eight hundred sixteen patients with chronic low back disability were included. All had a structured medical examination, including various physical tests before participation in either a functional restoration program (n = 621) or shorter "control" outpatient programs (n = 144). A smaller group of the cohort (n = 51) had no treatment and served as a pure control group. Six groups were selected from the cohort: Three underwent an identical functional restoration program and three underwent different outpatient control programs. Several baseline demographic, physical, and socioeconomic variables were correlated to 1-year outcome parameters. RESULTS: Age, days of sick leave, connection to the work force, and back pain intensity, were significantly correlated to success 1 year after entry into the study in all groups, no matter what kind of treatment was administered. Back muscle endurance, sports activity, activity of daily living scores, and vibrations were of importance in some outcome parameters for success after functional restoration. Smoking was positively correlated to disability pension. Days of sick leave and, in functional restoration, ability to work were the only factors that were correlative with statistics for people who withdrew. CONCLUSIONS: Different factors can be identified as predictive of outcome in a functional restoration program, but most of these factors were also shown to predict success for shorter control outpatient programs or of no treatment. PMID- 9728380 TI - Autologous bone grafting in staged scoliosis surgery. The patient as bone bank. AB - STUDY DESIGN: A prospective clinical study in which autologous rib graft, harvested during the thoracotomy in staged scoliosis correction, is stored within the patient for use during the second stage (posterior intrumentation and fusion). OBJECTIVE: To determine whether the bone stored by this technique is biologically viable and microbiologically safe. SUMMARY OF BACKGROUND DATA: To the authors' knowledge, this method of storage of bone has never been described previously. METHODS: During the first operation, the excised rib was divided into 3-5 cm fragments and stored in a sub-muscular plane adjacent to the posterior elements of the spine before closure. The graft was then retrieved at the second stage. Samples were sent for histologic and microbiologic examination before implantation. RESULTS: On histologic examination, more than 50% of the osteocytes retained their basophilic staining, indicating that they were viable. In addition, osteoclastic activity was notably absent. There was no significant bacterial contamination of the samples. Clinically, all patients achieved satisfactory bone fusion. CONCLUSION: Homeostatic equilibrium in humans provides the ideal environment in which bone graft can be stored. There is no increased risk of infection, and the osteogenic potential of the graft is retained. PMID- 9728379 TI - The transition zone above a lumbosacral fusion. AB - STUDY DESIGN: The clinical and radiographic effect of a lumbar or lumbosacral fusion was studied in 42 patients who had undergone a posterolateral fusion with an average follow-up of 22.6 years. OBJECTIVE: To examine the long-term effects of posterolateral lumbar or lumbosacral fusion on the cephalad two motion segments (transition zone). SUMMARY OF BACKGROUND DATA: It is commonly held that accelerated degeneration occurs in the motion segments adjacent to a fusion. Most studies are of short-term, anecdotal, uncontrolled reports that pay particular attention only to the first motion segment immediately cephalad to the fusion. METHODS: Forty-two patients who had previously undergone a posterolateral lumbar or lumbosacral fusion underwent radiographic and clinical evaluation. Rate of fusion, range of motion, osteophytes, degenerative spondylolisthesis, retrolisthesis, facet arthrosis, disc ossification, dynamic instability, and disc space height were all studied and statistically compared with an age- and gender matched control group. The patient's self-reported clinical outcome was also recorded. RESULTS: Degenerative changes occurred at the second level above the fused levels with a frequency equal to those occurring in the first level. There was no statistical difference between the study group and the cohort group in the presence of radiographic changes within the transition zone. In those patients undergoing fusion for degenerative processes, 75% reported a good to excellent outcome, whereas 84% of those undergoing fusion for spondylolysis or spondylolisthesis reported a good to excellent outcome. CONCLUSION: Radiographic changes occur within the transition zone cephalad to a lumbar or lumbosacral fusion. However, these changes are also seen in control subjects who have had no surgery. PMID- 9728381 TI - Anterior cervical fusion with the Orion locking plate system. AB - STUDY DESIGN: This study was conducted to evaluate an anterior cervical fusion plate system, the Orion locking plate, regarding its surgical handling, hardware related failures, and short-term and long-term results. OBJECTIVES: A comprehensive evaluation of the implant in a broad range of patients with cervical spine diseases. SUMMARY OF BACKGROUND DATA: Locking plates are the most recent devices for achieving anterior cervical spinal fusion and offer considerable advantages such as faster and easier implantation and fewer implant related failures than older plate systems. METHODS: Ninety-six patients were investigated. All underwent anterior cervical plate fusion as a component of the surgical treatment for symptomatic degenerative cervical spinal disease or for vertebral destruction caused by trauma, tumor, or inflammation. Besides plate fixation, 6 of the 96 patients had a combined ventrodorsal fusion. In 28 cases, one or more vertebral bodies were removed and replaced with titanium place holders. The remaining 62 patients were first treated by intervertebral inlay placement, and the fused segments were subsequently plated. Neurologic signs and symptoms were evaluated before and after surgery and during a follow-up period of at least 1 year. RESULTS: The rate of neurologic improvement was highest in radiculopathy patients and lowest in patients with severe myelopathy. In all cases, control radiographs demonstrated a solid bony fusion. Clinical deterioration after surgery was seen in four cases of severe myelopathy in which considerable neurologic deficits existed before surgery, possible because of rapid decompression of the cord and associated microvascular alterations. In two of these cases, there was long-term improvement. Additional general complications caused by surgical retraction included temporary swallowing disturbance in seven patients and a large wound hematoma in one. Hardware failures were encountered in three cases, all of them caused by improper implantation technique and not material failure, per se. CONCLUSION: In the study group, the Orion locking plate was easy to use, failure-free if properly implanted, safe for the patient and supported solid bony fusion in every case. PMID- 9728382 TI - Failure of a carbon fiber implant. A case report. AB - STUDY DESIGN: Case report. OBJECTIVES: Failure of a carbon fiber implant. SUMMARY OF BACKGROUND DATA: To simplify the procedure of posterior lumbar interbody fusion, a carbon-fiber-reinforced polymer implant has been developed. The implant has ridges to resist retropulsion, struts to support weight, and a hollow area to allow packing of autologous bone graft. So far, no complications have been reported from the use of carbon implant as a fusion aid in spine surgery. METHODS: A patient with postoperative infection has been followed with computed tomography images and histologic examination from a reoperation. RESULTS: An entire nonunion across the width of the disc space and a clearly broken cage was visualized with computed tomography. The spinal canal was explored during a reoperation and the tissue surrounding the dura and nerves were all black. Microscopic examination showed a large quantity of carbon particulate debris. The authors have operated on approximately 100 patients so far and no other carbon cage has broken, to their knowledge. CONCLUSIONS: Carbon cages can break if a nonunion occurs and as a result free carbon particles move out to the spinal canal. PMID- 9728383 TI - Bilateral cortical blindness after lumbar spine surgery. A case report. AB - STUDY DESIGN: A report of the unusual perioperative complication of bilateral cortical blindness after lumbar spine surgery. The hypothetical causes that can lead to this syndrome in spine surgery and the precautions are discussed. OBJECTIVES: The circumstances surrounding the occurrence of perioperative cortical blindness, the explanation of possible mechanisms, and the patients at risk are evaluated. SUMMARY OF BACKGROUND DATA: There have been no similar reports. METHODS: Case report with description of the syndrome of cortical blindness, the diagnostic tools, and the different pathophysiologic causes. RESULTS: The severe impairment of visual capacities remained unchanged; some color discrimination and the differentiation between dark and daylight were possible. CONCLUSIONS: In obese patients (body mass index > 30) puncture of the subclavian vein and rotating and positioning of the patient in one step should be performed carefully. PMID- 9728384 TI - Acute nontraumatic spinal epidural hematomas. An important differential diagnosis in spinal emergencies. AB - STUDY DESIGN: The clinical data of five patients with spontaneous spinal epidural hematoma (SSEH) were reviewed. OBJECTIVES: To assess the clinical outcome of patients with SSEH after surgical decompression. SUMMARY OF BACKGROUND DATA: The outcome in SSEH is essentially determined by the timing of the operation. Therefore, early and precise diagnosis is necessary. METHODS: A retrospective analysis of five patients with SSEH was performed. The clinical data were stratified according to the Frankel Score. Special interest was given to the relevance of rapid and exact diagnosis and immediate therapeutic intervention. RESULTS: Diagnosis of SSEH was established preoperatively by means of computed tomography (one case) or magnetic resonance imaging (three patients) and intraoperatively in one case. Lumbar myelography had been false negative in one patient, computed tomography false-negative in two patients. Surgical decompression was performed in four patients within 24 hours after the onset of symptoms. Favorable postoperative functional results were found only in one patient whose symptoms had been present for less than 12 hours and in the case of an incomplete cauda equina syndrome. CONCLUSIONS: The results of the current series demonstrate both the superiority of magnetic resonance imaging for diagnosis of SSEH as well as the necessity of early decompressive surgery in cases of sensorimotor paralysis after SSEH. PMID- 9728385 TI - Chylous leakage after circumferential thoracolumbar fusion for correction of kyphosis resulting from fracture: report of three cases. PMID- 9728386 TI - [Recent insights into the possibilities of resuscitation of dogs and cats]. AB - This article reviews the present state of the art of resuscitation of dogs and cats. The purpose of resuscitation is to revive animals so that the vital functions resume together with a normal brain function. Resuscitation must be started as soon as the cardiopulmonary arrest has been confirmed. Adequate ventilation and effective circulation to the most vital body organs, the heart and the brain, have the highest priority. They can be achieved by endotracheal intubation, artificial ventilation with 100% oxygen and rhythmic compression of the closed chest or direct cardiac massage following thoracotomy. Medical therapy is an important part of resuscitation. In the absence of a central venous route, deep endotracheal administration is the preferred method of administration. Most medications can be administered through the endotracheal tube in this fashion. PMID- 9728387 TI - [Infections with Baylisascaris procyonis in humans and raccoons]. AB - Baylisascaris procyonis is an ascarid which parasitizes the small intestine of raccoons. The parasite is not very pathogenic in the raccoon because larvae do not migrate in this host. In other animals the larvae migrate through the body. They do not develop into adult worms in the intestine but rather become encysted in granulomas, showing a preference for the brain. In humans these larvae cause different larva migrans syndromes. Patients with neural larva migrans syndrome show severe brain symptoms and the disease is sometimes fatal. This article describes the life cycle of the worm and the incidence, symptoms, diagnosis, treatment, and prevention of larva migrans syndromes, paying special attention to the Dutch situation. PMID- 9728388 TI - [Across individuals and species]. PMID- 9728389 TI - [Between depression and progress]. PMID- 9728390 TI - [Equine dental symposium. Veterinarians or assistants?]. PMID- 9728391 TI - [The veterinary disciplinary board]. PMID- 9728392 TI - [Position of the Royal Netherlands Veterinary Association concerning the union of veterinary pharmacists. Anything but defensive!]. PMID- 9728393 TI - Telomeres and double-strand breaks: trying to make ends meet. AB - Eukaryotic cells encounter two types of DNA ends: telomeres, the natural ends of linear chromosomes, and double-strand breaks, resulting from DNA damage or normal chromosomal processes such as meiotic or V(D)J recombination. These two termini have long been seen as functionally distinct, based on whether they are resistant to fusion with other ends or instead are acted upon by the DNA-repair machinery. However, a series of recent papers has shown that members of a set of proteins that are crucial for the rejoining of DNA strand breaks are also required for normal telomere function, raising new questions about how these two types of termini maintain their functional distinction. PMID- 9728394 TI - Time marches on. PMID- 9728395 TI - The control of filamentous differentiation and virulence in fungi. AB - Many members of the fungal kingdom have a distinguishing feature, dimorphism, which is the ability to switch between two morphological forms: a cellular yeast form and a multicellular invasive filamentous form. At least three pathways are involved in regulating the transition between these two forms in the budding yeast Saccharomyces cerevisiae, and evidence is now emerging that homologous signalling modules are involved in regulating filament formation and virulence in a range of human and plant fungal pathogens. Strikingly, components used to signal sexual differentiation in the response to mating pheromones are often reutilized to regulate dimorphic development, suggesting an ancient link between these processes. PMID- 9728396 TI - Vertebrate photoreceptor cell development and disease. AB - Photoreceptors provide an excellent model for studies of vertebrate neuronal differentiation, and many human diseases resulting in blindness primarily affect photoreceptors. There is therefore great interest in studying the cellular and molecular mechanisms of photoreceptor development. This article discusses our current understanding of this process, including the recent discovery of the homeodomain transcription factor Crx and its potential role in diseases affecting human vision. PMID- 9728397 TI - Integrin affinity modulation. AB - Integrin cell-adhesion receptors mediate interactions between cells and the extracellular matrix. Dynamic regulation of integrin adhesive function is termed 'activation' or 'inside-out' signalling. Activation is key to integrin function in processes as diverse as cell migration, the organization of the extracellular matrix and platelet aggregation. Consequently, there has been an intense effort to elucidate the molecular mechanism of integrin activation. This has resulted in the recent identification of novel cytoplasmic partners for integrins and the emerging characterization of the signal-transduction pathways that regulate integrin 'inside-out' signalling. Here, the authors review the recent developments that have provided us with an increased understanding of the basis of integrin activation. PMID- 9728399 TI - Lipids: more than just membrane fabric. PMID- 9728398 TI - Recognizing death: the phagocytosis of apoptotic cells. AB - Although apoptotic cell death is widespread, dying cells are rarely seen in situ because of their rapid clearance by neighbouring phagocytes. Phagocytic recognition of apoptotic cells is less well understood than the death programme itself, but an increasing number of recent studies are highlighting its importance. This review discusses the nature of the receptors that have been implicated in apoptotic cell phagocytosis, the mechanisms of uptake and the immunological consequences of apoptotic cell ingestion. PMID- 9728400 TI - Phenotypic and genotypic diversity of organisms previously classified as maltose positive Pasteurella multocida. AB - Fifteen isolates tentatively classified as maltose positive Pasteurella multocida have been characterized in 79 biochemical tests and by ribotyping using HindIII and HpaII for digestion of DNA. Phenotypic and genotypic results were analysed using the computer programmes NTSYS and GelCompar, respectively. Two strains were classified as maltose positive P. multocida ssp. multocida while six strains were classified as maltose positive P. multocida ssp. septica. The remaining strains clustered with P. volantium and P. gallinarum, but remained unclassified. With the exception of a single isolate correlation was observed between phenotypic and genotypic results. The unclassified isolates which represented three different sources were heterogenous according to both phenotypic and genotypic results. The findings obtained support the 16S rRNA sequencing results indicating that the genus Pasteurella sensu stricto might represent two or more genera. PMID- 9728401 TI - Influence of the phosphorylation state on the biological activity of a low molecular mitogen from group A streptococci. AB - A low molecular weight mitogen (LMP) from Streptococcus pyogenes strain NY 5 was successively purified by adsorption on phenylsepharose, chromatography on Resource S and Superdex G 30 and finally by affinity chromatography on antiphosphothreonine agarose. The N-terminal protein sequence of the mitogen was determined. The occurrence of phosphoamino acids was investigated by immunoassay using monoclonal antibodies. The LMP is a threonine-phosphorylated protein different of HPR protein of PTS-system, its mitogenic activity was lost after treatment with streptococcal protein phosphatase or alkaline phosphatase. The inactivated LMP was activated by phosphorylation with phosphokinase and ATP. The active LMP was also inactivated in streptococcal cultures secreting acid protein phosphatase during the phase of phosphate limitation. PMID- 9728402 TI - Serotaxonomic analysis of glycolipids from Mycobacterium chelonae-M. fortuitum complex and bovine farcy strains. AB - The antigenicity and cross-reactivity of glycolipids from strains of bovine farcy and the Mycobacterium chelonae-M. fortuitum complex were analyzed using the ELISA technique. Purified alkali-stable glycopeptidolipids (GPLs) with a characteristic dimethylrhamnosyl sugar unit extracted from M. abscessus, M. chelonae, M. peregrinum and M. senegalense, gave very strong reactions with sera against members of the same four species. Particularly strong cross-reactions were evident between M. peregrinum and M. senegalense. These GPLs reacted more weakly with antisera against the other mycobacteria tested, though clear reactions were noticed with M. farcinogenes and M. fortuitum and also with M. bovis BCG, M. phlei, and M. tuberculosis strains. Alkali-labile diacyl trehalose (DAT) and triacyl trehalose (TAT) from M. fortuitum reacted with homologous sera, and with that against M. tuberculosis. Traces of uncharacterized acyl trehaloses isolated from two strains of M. farcinogenes gave comparatively weak reactions. Mycobacteria labeled M. farcinogenes and M. senegalense produced glucosylated trehalose-based glycolipids (GTs) and the studies showed that the major type was antigenic. These glycolipids cross-reacted strongly with M. senegalense NCTC 4524 but not with the type strain of M. senegalense. On the basis of the chemical patterns and the antigenicity of the GPLs it is evident that M. peregrinum and M. senegalense are particularly closely related and these species show a very close affinity to M. abscessus-M. chelonae. PMID- 9728403 TI - Induction of apoptosis by Chlamydia psittaci and Chlamydia trachomatis infection in tissue culture cells. AB - The role of programmed cell death (apoptosis) in LLC-MK2 cells infected with Chlamydia trachomatis LGV2 serotype and Chlamydia psittaci 6BC strain was investigated using flow cytometry and TUNEL procedures. The number of apoptotic cells was significantly higher at 72 and 96 hours post infection in the Chlamydia infected cell cultures in comparison with mock-infected cells. We postulate the apoptotic process to be a mechanism induced by C. trachomatis and C. psittaci infection in LLC-MK2 cells. PMID- 9728404 TI - Identification of group VS116 strains among Borrelia burgdorferi sensu lato grown from the hard tick, Ixodes ricinus (Linnaeus, 1758) by off-coupled restriction fragment length polymorphism analysis. AB - A total of 67 spirochetal isolates grown from the hard tick, Ixodes ricinus were analysed by PCR amplification of the spacer region between two conserved structures, the 3' end of the 5S rRNA and the 5' end of the 23S rRNA genes of Borrelia burgdorferi sensu lato. A 246-255 bp amplicon was generated from 13 reference strains of B. burgdorferi sensu lato representing the three major genospecies, B. burgdorferi sensu stricto, B. garinii, and B. afzelii, and from all 67 spirochetal isolates from ticks but not from B. hermsii. As could be confirmed by DNA sequence analysis, restriction fragment length polymorphism (RFLP) analysis of the PCR product after cleavage with DraI and MseI distinguished between the three major genospecies: out of the 67 B. burgdorferi sensu lato isolates from ticks, 27 (40.3%) were typed as B. burgdorferi sensu stricto including five isolates with a unique DraI or MseI pattern. 26 isolates (38.8%) were typed as B. garinii and 6 (9.0%) as B. afzelii, respectively. A group of eight isolates (11.9%) displayed a unique MseI pattern identical to that described for a putative new European genospecies of Borrelia burgdorferi sensu lato designated VS116. DNA sequences of the PCR product of seven of these isolates tested were by less than 88.5% identical with the established European major genospecies but shared 98% to 100% homology with that of database-derived sequences of strain VS116 from Switzerland and strain UK from England which are both representatives of the European genomic group VS116. PMID- 9728405 TI - Evaluation of ligase chain reaction for direct detection of Mycobacterium tuberculosis in respiratory specimens. AB - A new automated amplification method, Ligase Chain Reaction (LCx MTB), was evaluated for direct detection of Mycobacterium tuberculosis in respiratory specimens from 208 patients and its performance was compared with culture and direct smears. Out of 226 specimens, 28 LCx MTB and 15 cultures were found positive for M. tuberculosis. After resolution of clinical history, the sensitivity of LCx MTB and culture was respectively 89.3% and 53.6% with a specificity of 98.5% and 100%. However, samples coming from untreated patients presented similar results between culture and LCx MTB (sensitivity 75% and 83.3% for culture and LCx MTB). PMID- 9728406 TI - Biochemical characteristics, serogroup distribution, antibiotic susceptibility and age-related significance of Campylobacter strains causing diarrhoea in humans in Hungary. AB - During August and September 1995, 111 thermopilic campylobacters from stool samples of patients suffering from diarrhoea were cultured and f1amined. Biochemical characteristics, serological distribution and antibiotic susceptibility of the strains were examined and the age distribution of the patients affected was recorded. Most of the strains, i.e. 101 isolates (91%) proved to be Campylobacter (C.) jejuni, whereas 10 strains (9%) were C. coli. On the basis of their heat-stable antigens, 66 strains (65.3%) of C. jejuni could be assigned to 17 serogroups, of which serogroups 2 (15 strains, 14.8%) and 8 (10 strains, 9.9%) occurred most frequently. All isolates examined were susceptible to erythromycin whereas susceptibility to other antibiotics varied greatly. Children under five years of age (59 cases = 53.1%) were most frequently affected. During 1995, altogether 11,976 human Campylobacter cases were recorded in Hungary which means a prevalence of 114/100,000. The results suggest that the great majority of cases of Campylobacter diarrhoea is caused by C. jejuni strains while C. coli strains have much less significance. The serotype distribution of C. jejuni strains causing diarrhoea is very wide. If treatment is needed the best choice at present seems to be erythromycin. PMID- 9728407 TI - Transfer of resistance to oxy-imino-cephalosporins and of extended-spectrum beta lactamase productions in Klebsiella pneumoniae strains from infected neonates. AB - In this communication, we describe the occurrence of strains of Klebsiella pneumoniae resistant to cephalosporins of all generations and to aztreonam due to their production of Extended Spectrum beta-Lactamases (ESBLs), in two hospitals in Slovakia. They were found to transfer the genetic determinants of resistance of cefotaxime, ceftazidime and aztreonam and of ESBL production to suitable recipient strains of Escherichia coli K-12 No. 3110 and Proteus mirabilis P-38. Six donor K. pneumoniae strains were collected from six prematurely born babies gradually infected with strains of K. pneumoniae resistant to cefotaxime, ceftazidime and aztreonam. All six strains of K. pneumoniae gave an identical pattern in ESBL testing (double-disk diffusion test). The second cycle of transfers to nalidixin-resistant E. coli K-12 No. 185 nal+ also confirmed that all transconjugants were resistant to all beta-lactams tested. We conclude that a single gene was transferred from donor strains which confers 'en bloc' the resistance to the beta-lactams tested. Four strains of K. pneumoniae produced a uniform type of ESBI. Although with different quantitative expression. All transconjugants tested produced identical types of ESBL as did the donor strains of K. pneumoniae. This transfer of an identical pattern of ESBL production was confirmed also in the second cycle of transfers. Cefotaxime, ceftazidime and aztreonam were actively hydrolysed (as shown by the relative rate of hydrolysis [Vmax]) by strains of K. pneumoniae as well as by transconjugant colonies and clavulanate inhibited the hydrolysis of these beta-lactam antibiotics. PMID- 9728408 TI - Actinobacillus actinomycetemcomitans in the human oral microflora. AB - In this paper, the frequency of the microaerophilic gram-negative bacterium Actinobacillus actinomycetemcomitans (Ac) which is a component of the normal human oral microflora was investigated. Ac could be cultivated from oral material (molar sulcus and/or mucous membrane of the cheek) from 55 out of 405 healthy adults examined. This overall Ac frequency of 13.6% corresponds well to the few other reports existing in literature. A short discussion about the possible pathogenic potential of Ac is given at the end of the paper. PMID- 9728409 TI - Putative origin of Yersinia pseudotuberculosis in western and eastern countries. A comparison of restriction endonuclease analysis of virulence plasmids. AB - Yersinia pseudotuberculosis isolates from Russia east of Moscow, Korea and mainland China were used for restriction endonuclease analysis of virulence plasmid (REAP) and findings were compared with REAP of isolates from Japan and Western countries. An identical REAP pattern of each serogroup 1a, 1b, 3, 4a and 4b strain was observed among isolates from Russia, Korea, mainland China, and Japan but such was absent in West European strains. Therefore, the possibility that the origin of Y. pseudotuberculosis between West Europe and eastern Eurasia east of Moscow may be from a different clone should be considered. PMID- 9728410 TI - Killing of Pseudomonas pseudomallei by polymorphonuclear leukocytes and peritoneal macrophages from chicken, sheep, swine and rabbits. AB - Differences in the kinetics of Pseudomonas pseudomallei killing by peritoneal macrophages (PM) and polymorphonuclear leucocytes (PMNL) from chickens, sheep, swine and rabbits were found. P. pseudomallei was rapidly killed by porcine PM and PMNL. However the bacterial killing by ovine and lapine PM and PMNL proceeded at a slower rate. In contrast, chicken PM and PMNL ingested and killed the lowest number of P. pseudomallei bacteria. The differences in the bactericidal activity of PM and PMNL from different animal species correlated with the level of their acid phosphatase and glycolytic activity. PMID- 9728411 TI - Clinical features of Lyme borreliosis in the middle Urals and distribution of Borrelia burgdorferi sensu lato species in local Ixodes persulcatus ticks. AB - From 1991 to 1993, we investigated the clinical features of Lyme borreliosis (LB) in 864 patients from the Sverdlovsk region (population 4.5 million) in the middle Urals. Ixodes persulcatus ticks were collected in the vicinity of Yekaterinburg to determine the presence of Borrelia burgdorferi sensu lato species. From 1991 to 1993, the number of patients with LB increased from 91 to 320 and 453, respectively. Nearly all LB patients (97%) recalled a tick bite and the first signs of LB developed between May and August. Erythema migrans (EM) was seen in 820 patients (94.9%) and fever was common (44.6%). Neuroborreliosis, mainly radiculoneuritis, was found in 154 patients (17.8%), secondary erythema was seen in 53 patients, and Lyme arthritis (LA) was diagnosed in 35 patients. Carditis was rare and acordermatitis chronica atrophicans (ACA) was not seen. Only 44 patients developed one or more symptoms of LB without a preceding EM. Few patients were seropositive for tick-borne encaphalitis. Borrelial DNA was detected in 67% of I. persulcatus ticks. Infected ticks carried predominantly Borrelia garinii or Borrelia afzelii, mixed infections of both species were common. Borrelia valaisiana was detected once, and B. burgdorferi sensu stricto was not found. Although the course of LB in the middle Urals in comparable to that of European LB, several discrepancies were noted. LB patients often recall tick bites, fever is common, LA is mild and ACA is absent. PMID- 9728412 TI - The complement-killing of Borrelia burgdorferi. Target antigens and sensitizing antibodies. AB - It had been previously shown by the Microbial Adherence Immobilization Assay (MAIA) that Borrelia burgdorferi sensu stricto, type strain B31 was clumped, immobilized and killed in vitro by sensitizing antibodies that activated the classical complement pathway and the complement-killing of live borrelia. In the present study, the target antigens and sensitizing antibodies responsible for the complement-killing of borrelia were investigated, using MAIA as a selective identification tool. It was found that the fractions containing the 31 and 34 kDa outer surface proteins from strain B31 were the unique antigens producing sensitizing antibodies in rabbits that activated the complement-killing of B31. An anti-OspB, but not an anti-OspA, monoclonal antibody did activate the B31 complement-killing in MAIA. From these results, constraints on the effectiveness of OspB and OspA as immunogens for the prevention and control of Lyme borreliosis in humans are discussed. PMID- 9728413 TI - Detection and characterization of two cytotoxins produced by Campylobacter jejuni strains. AB - Campylobacter jejuni strains are able to produce at least two different cytotoxins called "cytolethal distending toxin" (CLDT) and "cytolethal rounding toxin" (CLRT). In this study, we investigated the corresponding changes in CHO-K1 cells using the cell counter and analyzer system CASY 1. Determination of the cell volume after toxin treatment of the cells is a useful criterion for differentiation between the cytotoxic activities produced by Campylobacter strains. Incubation of the cells with crude CLDT resulted in a decrease in the cell count combined with a dramatic increase of the mean cell volume in comparison to the control culture. A decrease in the cell count was also seen as a response to CLRT preparations, while this toxin had no effect on the mean cell volume determined. It was shown that only CLDT caused histone-associated DNA fragments in the cytoplasm of CHO-K1 cells indicating an apoptotic pathway of cell death. In addition, the polymerase chain reaction (PCR) was employed to screen Campylobacter strains for the presence of the cdtB gene sequence, which was detectable in all strains investigated. PMID- 9728415 TI - What is beyond the big five? AB - Previous investigators have proposed that various kinds of person-descriptive content--such as differences in attitudes or values, in sheer evaluation, in attractiveness, or in height and girth--are not adequately captured by the Big Five Model. We report on a rather exhaustive search for reliable sources of Big Five-independent variation in data from person-descriptive adjectives. Fifty three candidate clusters were developed in a college sample using diverse approaches and sources. In a nonstudent adult sample, clusters were evaluated with respect to a minimax criterion: minimum multiple correlation with factors from Big Five markers and maximum reliability. The most clearly Big Five independent clusters referred to Height, Girth, Religiousness, Employment Status, Youthfulness and Negative Valence (or low-base-rate attributes). Clusters referring to Fashionableness, Sensuality/Seductiveness, Beauty, Masculinity, Frugality, Humor, Wealth, Prejudice, Folksiness, Cunning, and Luck appeared to be potentially beyond the Big Five, although each of these clusters demonstrated Big Five multiple correlations of .30 to .45, and at least one correlation of .20 and over with a Big Five factor. Of all these content areas, Religiousness, Negative Valence, and the various aspects of Attractiveness were found to be represented by a substantial number of distinct, common adjectives. Results suggest directions for supplementing the Big Five when one wishes to extend variable selection outside the domain of personality traits as conventionally defined. PMID- 9728414 TI - Epidemiological typing of Alcaligenes xylosoxidans subsp. xylosoxidans by antibacterial susceptibility testing, fatty acid analysis, PAGE of whole-cell protein and pulsed-field gel electrophoresis. AB - Antibacterial susceptibility testing, fatty acid analysis, protein analysis and DNA analysis of Alcaligenes xylosoxidans subsp. xylosoxidans were compared to determine the efficiency of the methods available for strain typing. Thirty isolates were investigated: 20 clinical isolates from a nonsocomial outbreak in Essen (Germany), 9 clinical isolates from sporadic nosocomial cases in Paris (France) and reference strain ATCC 2402. The highest microbiological discriminative power was exhibited by pulsed-field gel electrophoresis (PFGE) yielding nine types, followed by fatty acid methyl ester (FAME) analysis with six types, and antibacterial susceptibility testing and polyacrylamide gel electrophoresis with five types each. By combining the results of the four typing methods, 14 varieties could be differentiated. Protein analysis and fatty acid analysis failed to discriminate between isolates from Essen and Paris and the reference strain, while antibacterial susceptibility testing and DNA analysis clearly discriminated them. It is concluded that a combination of antibacterial susceptibility testing and PFGE typing is most suitable for epidemiological typing of Alcaligenes xylosoxidans subsp. xylosoxidans strains. PMID- 9728416 TI - Shame, guilt, ego development, and the five-factor model of personality. AB - Shame, guilt, and ego development are conceptually interrelated constructs, yet their empirical relations have not yet been examined. Further, these constructs have not yet been mapped onto the widely used Five-Factor Model. In Study 1, relations were examined between these three domains within a sample of Australian university students. Two types of guilt were distinguished, Empathic Guilt (associated with Agreeableness) and Anxious Guilt (associated with Neuroticism). The relationship between Shame and Ego Level was found to be curvilinear, with Shame greatest for persons at intermediate stages of ego development. In Study 2, relations between ego development and the Five-Factor Model were further examined. Across both studies, Ego Level was best predicted from Conscientiousness among men and from Openness among women. Relations between Ego Level, proneness to shame and guilt, and the five factors were typically modest, suggesting that these represent complementary approaches to the study of personality. PMID- 9728417 TI - Primary and secondary control over age-related changes in physical appearance. AB - Beliefs about appearance-related changes due to aging were used to test the effects of perceived control and secondary control (acceptance) in a sample of 412 young, early-middle-age, and late-middle-age college-educated adults. Mean difference in aging-related appearance control and hypotheses regarding the adaptiveness of primary and secondary control were examined. Primary control over aging-related appearance was lower in older adults and secondary control was higher. In addition, the results indicated support for the Primacy/Back-Up Model that primary perceived control is important at all levels of actual control. Those with stronger beliefs in their primary control were less distressed. Secondary control served a back-up function in that it was related to less distress only for those who had medium or lower beliefs in primary control. The implications of these findings, that primary control may be advantageous even in low-control circumstances, are discussed. PMID- 9728418 TI - Generalization, adverse events, and development of depressive symptoms. AB - Many diathesis-stress models have been proposed in which cognitive processes of various types are presumed to represent vulnerabilities to development of depressive symptoms. This study tested three potential vulnerabilities as prospective predictors of such symptoms: the holding of especially high standards, the tendency to be self-critical after failure, and the tendency to generalize from a single failure to the broader sense of self-worth. At the start of a semester, college students completed a measure of these cognitive tendencies and a measure of depressive symptoms. Six weeks later they completed the same measure of depressive symptoms and a brief measure of intervening life events. Hierarchical regression analysis yielded evidence that Generalization interacted with adverse events to predict subsequent depressive symptoms. Self-Criticism also tended to predict later symptoms, but only if the symptoms were present initially. High Standards had no adverse effect. PMID- 9728419 TI - Prognostic value of prostate specific antigen before, during and after radiotherapy. PMID- 9728421 TI - Medical management of advanced gastric cancer. PMID- 9728420 TI - Clinical, pathological and therapeutic aspects of oligodendroglioma. PMID- 9728423 TI - Chemotherapy in advanced pancreatic adenocarcinoma. PMID- 9728422 TI - Colorectal cancer: treatment of advanced disease. PMID- 9728424 TI - Clinical implications of molecular and cytogenetic studies of non-Hodgkin's lymphomas. PMID- 9728426 TI - The endovascular management of symptomatic atherosclerotic carotid disease. PMID- 9728425 TI - The significance of one occluded internal carotid artery. PMID- 9728427 TI - Nitric oxide and renal reperfusion injury: a review. AB - BACKGROUND: Nitric oxide (NO) is a ubiquitous molecule with a role in both normal homeostasis and pathophysiological processes. However, the role it plays in renal ischaemia reperfusion injury is matter of some contention. METHOD AND RESULTS: This paper precis the biology and biosynthesis of NO before concentrating on the function of NO in renal ischaemia reperfusion injury. What emerges is a picture of some clarity about the normal physiological role of NO but some discord regarding the generation and function of postischaemic nitric oxide. Some of the reasons for this are explored in more detail. CONCLUSIONS: Clarity on the precise effect of postischaemic NO will remain elusive without the in vivo measurement of NO in experimental models. PMID- 9728428 TI - The fate of patients undergoing surveillance of small abdominal aortic aneurysms. AB - OBJECTIVES: Increasing numbers of patients with small abdominal aortic aneurysms (AAA) are being diagnosed. The aim of this paper is to define the fate of those patients undergoing surveillance of small AAAs. SETTING: U.K. district general hospital. METHODS: A prospective study has been carried out of all patients undergoing surveillance. At the time of the first consultation the patient was assessed, a Detsky score calculated and the referral source noted. End points of the study were elective repair of the aneurysm, aneurysm rupture or death of the patient. RESULTS: Details of 267 patients were analysed. The referral source was general practitioner in 39%, patients with peripheral vascular disease in 32% and department of urology in 21%. None were referred from population screening. The cumulative 5-year risks of rupture, elective repair or non-AAA related deaths were 15%, 26% and 46% for all patients, 4%, 13% and 38% for patients initially presenting with AAA less than 4 cm diameter and 21%, 42% and 54% for patients presenting with an AAA 4-5.5 cm diameter. All but one of 11 patients whose aneurysm ruptured were unfit or had declined elective repair. There were 56 non AAA related deaths, the majority due to cardiovascular causes. Those patients with low Detsky scores had a 5-year survival of 62%, those with high scores 44%. The age/sex matched survival or a normal population at 5 years in 80%. CONCLUSION: Overall the non-AAA related mortality was greater than the risks of rupture or elective repair. It is important to bear in mind the poor prognosis of this group of patients compared with a normal population when considering elective repair of small AAAs. PMID- 9728429 TI - Intravascular ultrasound predictors of restenosis after balloon angioplasty of the femoropopliteal artery. AB - OBJECTIVES: To determine intravascular ultrasound parameters related to restenosis following percutaneous transluminal balloon angioplasty (PTA) of the femoropopliteal artery. DESIGN: Prospective study. MATERIALS AND METHODS: Patients were studies with intravascular ultrasound before and after angiographic successful PTA (n = 114). Intravascular ultrasound cross-sections obtained with 1 cm interval in the dilated segment were analysed. A distinction was made between anatomic (duplex scanning) and clinical (Rutherford criteria) restenosis assessed within 1 month and at 6 months after PTA. RESULTS: Intravascular ultrasound predictors of 1 month anatomic outcome were lumen area stenosis after PTA, lumen area increase, plaque area decrease, and area stenosis decrease; predictor of 6 months anatomic outcome was area stenosis after PTA. Multivariate analysis revealed that area stenosis after PTA was the only independent predictor of both 1 and 6 months anatomic outcome. Intravascular ultrasound predictors of 1 month clinical outcome were the presence of hard lesion and the mean arc of hard lesion. Multivariate analysis revealed that the mean arc of hard lesion was the only independent predictor of 1 month clinical outcome. No predictors for 6 months clinical outcome were found. CONCLUSIONS: Intravascular ultrasound can elucidate parameters predictive of restonosis after PTA. The strongest intravascular ultrasound parameter predictive of anatomic restenosis was a large area stenosis after PTA. PMID- 9728430 TI - The feeling of loneliness prior to coronary artery bypass grafting might be a predictor of short-and long-term postoperative mortality. AB - OBJECTIVES: To evaluate the effect of different aspects of quality of life (QL) upon mortality during short-and long-term follow-up after coronary artery bypass grafting (CABG). DESIGN: Prospective evaluation. MATERIALS: Consecutive patients from western Sweden who during 3 years underwent CABG. METHODS: They answered a questionnaire at the time of coronary angiography prior to CABG. Quality of life was measured with questions from the Nottingham Health Profile (NHP) part I. RESULTS: In all, 1290 patients were included in the analyses. When accounting for various preoperative factors known to be independently associated with morality the NHP question "I feel lonely" was found to be associated with mortality, both at 30 days (RR 2.61; 95% CI 1.15-5.95; p = 0.02) and at 5 years (RR 1.78; 95% CI 1.17-2.71; p = 0.007) after the operation. Thirteen per cent reported they felt lonely. At 5 years was, in addition, the statement "I have difficulty climbing stairs" also independently associated with mortality (RR 1.50; 95% CI 1.02-2.22; p = 0.04). CONCLUSION: Among the 38 statements in NHP as a judgment of QL prior to CABG, one of them, "I feel lonely" was independently associated with survival both at 30 days and 5 years after CABG. PMID- 9728432 TI - Duplex scanning and CT angiography in the diagnosis of carotid artery occlusion: a prospective study. AB - OBJECTIVES: Differentiating total occlusion from tight stenosis of the internal carotid artery is crucial with regard to treatment and prognosis. At our institution, the diagnosis of carotid stenosis is based on duplex scanning. In cases of occlusion, duplex is not reliable, and angiography is performed, thereby increasing morbidity. We tried to determine whether a combination of duplex scanning and CT angiography (CTA) can replace angiography in the diagnosis of carotid occlusion. DESIGN: Prospective study. MATERIALS AND METHODS: From 1995 to 1997, 148 patients were diagnosed as having carotid occlusion by duplex scanning. CTA was performed on all patients. Forty-four patients underwent angiography and 10 patients were surgically explored. Both procedures were considered "gold standard" for the diagnosis of occlusion. RESULTS: Arteries found to be occluded by both CTA and duplex scan were confirmed as occluded by angiography or operation in 95% of the cases (42/44). Arteries found to be occluded by duplex but patent by CTA were confirmed as patent in 100% of cases (10/10). CTA has a significantly higher positive predicting value for diagnosing occlusion than duplex scan (95% vs. 77%, p value < 0.01). CONCLUSIONS: Combination of duplex scanning and CTA is safe and accurate in the diagnosis of carotid occlusion and can replace angiography in most cases, thereby reducing morbidity. PMID- 9728431 TI - A comparison between PTFE and denatured homologous vein grafts for haemodialysis access: a prospective randomised multicentre trial. The SMASH Study Group. Study of Graft Materials in Access for Haemodialysis. AB - OBJECTIVES: To compare patency and complication rates of polytetrafluoroethylene (PTFE) grafts and denatured homologous vein (DHV) grafts for long-term haemodialysis. DESIGN: A prospective randomised multicentre trial. MATERIALS: One hundred and thirty-one patients were enrolled between September 1994 and April 1997. Sixty-three DHV grafts and 68 PTFE grafts were implanted in 60 meals and 71 females. Complications and interventions were monitored. Patency rates, complication rates, and intervention rates of PTFE and DHV were compared. RESULTS: The mean follow-up was 313 days for DHV (range 1-771) and 339 (3-909) days for PTFE. The total follow-up was 54.1 patient-years for DHV and 63.1 for PTFE. The 1-year primary patency rates were 30% and 40% for DHV and PTFE respectively. Secondary patency rate was 63% for both DHV and PTFE. Most frequent complication was thrombosis. A total of 75 thrombotic events (1.39 per patient year) occurred in 35 (56%) DHV grafts and 78 (1.24 per py) in 36 (53%) PTFE grafts. A total of nine infections were seen in nine (14%) DHV grafts, whereas 21 infections in 20 (29%) PTFE grafts were seen (p = 0.08). All but one infected DHV graft could be salvaged with systemic antibiotics. In contrast, surgical intervention was necessary in nine PTFE grafts (p = 0.02). For aneurysms, eight DHV and two PTFE grafts needed revision (p = 0.03). CONCLUSION: Patency rates between DHV and PTFE were not different. More infections were seen in PTFE grafts, and significantly more PTFE grafts needed surgical revision or removal because of infection. Significantly more DHV grafts were surgically revised or removed because of aneurysms. PMID- 9728433 TI - Outcome and influence of age after infrainguinal revascularisation in critical limb ischaemia. The Swedish Vascular Registry. AB - OBJECTIVES: To evaluate whether revascularisation has any influence on the mortality rate, and the impact of old age in patients with critical limb ischaemia (CLI). DESIGN: Analysis of Swedish Vascular Registry (Swedvasc) data. PATIENTS AND METHODS: During 1987-1995, 3730 surgical and 1199 endovascular (PTA) procedures below the groin due to CLI were reported. At 1 year three groups were defined: "occluded, amputated"; "occluded, not amputated" and "patent". Survival was also calculated. Clinical outcome at 1 month and at 1 year was defined as: patient "alive, improved", "alive, not improved", "alive, amputated" and "dead". Two age groups < or = 75 years or and > or = 76 years were compared. RESULTS: The mortality rate for the whole group was 5.3% at 1 month and 22.9% at 1 year, with no difference between the Surgery and PTA groups. Significantly more patients were alive and improved after surgery than after PTA at 1 month (82.3% vs. 77.7%) and at 1 year (49.6% vs. 44.3%). The amputation rate was 5.6% at 1 month and 14.4% at 1 year; 17% for diabetics. After surgery, the cumulative mortality rate did not differ between patients with a salvaged limb, irrespectively of patency of the re-construction, but was significantly higher after amputation. After PTA, only a reconstruction reported as patent was linked to the most favourable survival rate. The older patient group had a mortality rate of 6.4% at 1 month and 26.4% at 1 year, significantly higher than the younger group (3.8% and 17.6%, respectively). The amputation rate did not differ according to age. Significantly more patients were alive but not improved in the older group. CONCLUSIONS: The outcome of surgery vs. PTA was similar regarding survival and amputation, but surgery resulted in a greater improvement although this might be due to selection. Older patients and those with an amputation had higher mortality rates. PMID- 9728434 TI - Evaluation of the risks of using an oversized balloon catheter in the human infrarenal abdominal aorta. AB - OBJECTIVES: To evaluate the effects on the aortic wall of balloon dilatation as utilised in deployment of stent-graft devices during endoluminal repair of infrarenal abdominal aortic aneurysm. METHODS: Large dilatation balloons were expanded within the aorta of 41 cadavers. Testing was done to evaluate the effect of differing degrees of balloon oversizing, at pressures in the range of 0.15-2.5 atm. The aorta was then open for macroscopic inspection. RESULTS: In group 1 (mild atherosclerosis) no macroscopic abnormalities were detected with up to 6 mm oversized balloon. In group 2 (moderate atherosclerosis) fracture of atherosclerotic plaque occurred in seven of 14 aortas (50%) with 2.5 mm-4mm oversized balloon. In group 3 (severe atherosclerosis) fracture of atherosclerotic plaque occurred in six of seven (85%) with 2.5 mm to 4 mm oversized balloon and rupture of the aorta occurred at 6 mm oversizing. CONCLUSIONS: This study suggests that balloon overdilatation of the aorta by 2 mm, at pressures less than 2 atmospheres, allows safe deployment even in the presence of severe atheroma. Larger amounts of overdilatation are relatively safe in mildly atherosclerotic aorta. Aortic rupture is unlikely with overdilatation up to 6 mm, especially in less calcified vessels. PMID- 9728435 TI - A clinical and haemodynamic investigation into the role of calf perforating vein surgery in patients with venous ulceration and deep venous incompetence. AB - OBJECTIVE: To determine the clinical efficacy and local haemodynamic effects of perforating vein surgery in ulcerated limbs with combined deep and perforating vein incompetence. DESIGN: Prospective, interventional study. MATERIALS AND METHODS: Seven ulcerated limbs with combined primary deep and perforating vein incompetence were studied. Clinical efficacy was determined by ultimate ulcer healing and reduction in ulcer area, local haemodynamics were assessed at three sites with photoplethysmographic 90% venous refilling times (PPG RT90); both assessments were performed pre- and 1-month postoperatively. RESULTS: None of the ulcers healed following perforating vein surgery, the median (range) ulcer areas pre- and postoperatively were 31 (7-685) cm2 and 35.5 (7-796) cm2 (Wilcoxon p = 0.07). Preoperative PPG RT90 demonstrated a global abnormality of venous function at all sites examined that persisted after perforating vein surgery. CONCLUSION: In the presence of deep venous incompetence perforating vein surgery had no influence on venous function or ulcer healing. We conclude that perforating vein surgery is not indicated for the treatment of venous ulceration in limbs with primary deep venous incompetence. PMID- 9728436 TI - Intraoperative duplex scanning for carotid endarterectomy. AB - OBJECTIVES: To evaluate the results of intraoperative duplex scans during carotid endarterectomy. DESIGN: Retrospective case review. MATERIALS: One-hundred consecutive intraoperative carotid duplex scans performed during carotid endarterectomy between July 1993 and December 1995 at a university teaching hospital. METHODS: Abnormalities of the B-mode image and/or the Doppler flow analysis were classified. The result of intraoperative carotid duplex scans (ICDS) were related to the events of the intraoperative course, perioperative neurologic morbidity and mortality, and to residual carotid stenosis. RESULTS: Abnormalities of the ICDS were demonstrated in 13 cases (13%). Abnormalities were classified into four types: I, internal carotid artery spasm (n = 9); II, high distal resistance flow (n = 2); III, high grade residual stenosis (n = 1); IV, intraluminal thrombosis (n = 1). Immediate intraoperative exploration and revision of the endarterectomy was performed based on the ICDS in two cases (type III and IV) and the findings of ICDS were confirmed. The other 11 cases with abnormal ICDS (types I, II) were not revised and duplex scans done 1 month postoperatively (available in 10 cases) showed normal carotid artery flow. Intraoperative angiography was performed selectively in five cases and confirmed the results of ICDS. Reversible abnormalities of the ICDS were not associated wit perioperative morbidity or residual carotid stenosis. CONCLUSIONS: Intraoperative carotid duplex scanning can be used to assess the immediate technical adequacy of carotid endarterectomy. B-mode image and Doppler flow abnormalities which are reversible can be distinguished from those which require immediate revision. PMID- 9728437 TI - Results of surgery and angioplasty for the treatment of chronic severe lower limb ischaemia. AB - OBJECTIVE: To aims of this study was to assess and compare the efficacy of PTA and surgery in the treatment of severe lower limb ischaemia. DESIGN: Prospective 12-month study of 180 consecutive patients with severe chronic lower limb ischaemia. METHODS: PTA was used as first line therapy whenever possible and appropriate. Surgical revascularisation, primary amputation and conservative therapy were used in eh remaining patients. Patient survival and limb salvage were derived using life table analysis. RESULTS: Revascularisation was attempted in 135 (75%) patients, with PTA in 82 (46%), surgery in 19 (27%) and a combination of both in four (2%). Overall 12-month survival and limb salvage was 75% and 71%, respectively. Surgery and PTA had significantly higher survival rates (91% and 78%) than primary amputation or conservative therapy (57% and 52%) (p < 0.0001 log rank test). Revascularisation with either surgery or PTA achieved the same limb salvage rate of 76%. CONCLUSION: A large proportion of patients with severe chronic lower limb ischaemia can be managed by PTA. THis management strategy produces a clinically effective outcome at 1-year. PMID- 9728438 TI - Obturator bypass to the distal profunda femoris artery using a medial approach- long-term results. PMID- 9728439 TI - Acute exercise and markers of endothelial injury. PMID- 9728440 TI - Exercise in patients with intermittent claudication. PMID- 9728441 TI - Carotid surgery. PMID- 9728442 TI - Hyperhomocysteinaemia. PMID- 9728443 TI - Rifampicin-soaked grafts. PMID- 9728444 TI - Does MDT arrest transmission of leprosy to household contacts? AB - The multidrug therapy program with the World Health Organization (WHO) recommended treatment (WHO/MDT) regimens has given the hope of early case detection and rendering a leprosy patient, especially a multibacillary (MB) patient, noninfectious within a short period of time. Hence, the duration of exposure for household contacts to infection is expected to be remarkably less when compared to exposure to MB leprosy patients on dapsone monotherapy. A total of 1661 household contacts of skin-smear-positive leprosy patients were recorded from 1984 to 1994. Follow up of these individuals [8403 person-years at risk (PYR)] revealed that the incidence of leprosy was 7.7 per 1000 PYR, which was 8 times more than that of the general population. The risk was more if there was a coprevalent case in the family. The incidence of leprosy declines from the third year of surveillance onward, and declines more so in children. Although disease transmission should have been arrested as soon as the index case was started on MDT, the incidence of leprosy among the household contacts was still high when compared to that of the total population. Effective intervention needs to be introduced to reduce the risk of contacts developing leprosy. PMID- 9728445 TI - Epidemiological significance of first skin lesion in leprosy. AB - The epidemiological significance of monolesions in leprosy and the possible inferences on the mode of entry by Mycobacterium leprae into the body are presented based on data from the clinical records of the Leprosy Control Programme of Gudiyatham Taluk in India; 660 children with monolesions (335 males, 305 females) younger than 15 years of age and detected during the period 1990 1995 were included in the study. Detailed investigations on the location of monolesions were carried out and compared with a random sample of 669 normal rural children matched for age and sex. A large majority of the leprosy monolesions were in the uncovered parts of the body, with special predilection for the posterior aspects of the upper extremities and the anterior aspects of the lower extremities. Based on observation of normal children, these happen to be precisely the sites vulnerable for trauma since they are exposed to the environment where M. Leprae could enter through abraded skin and manifest as a patch. The need for further studies is emphasized. PMID- 9728446 TI - Spontaneous leprosy in a wild-caught cynomolgus macaque. AB - Naturally occurring Mycobacterium leprae has been previously documented in only two species of nonhuman primates from West Africa--the chimpanzee and the sooty mangabey. We report here the first known case of spontaneous leprosy in an Asian macaque. A wild-caught cynomolgus macaque imported from The Philippines developed a reaction to a tuberculin skin test after 3 years at the California Regional Primate Research Center (CRPRC), University of California-Davis, Davis, California, U.S.A. Biopsies of concurrent skin lesions suggested a cutaneous mycobacterial infection. Diagnosis of the infection was obtained by a polymerase chain reaction (PCR) assay specific for M. leprae. Clinical presentation, histopathological findings, and ELISA serology for M. leprae-specific PGL-I and to the LAM mycobacterial antigens were consistent with those of human borderline (BB) leprosy. Longitudinal serologic data suggest that the cynomolgus macaque had subclinical leprosy at the time of arrival in the CRPRC quarantine. Intradermal tuberculin testing is the traditional method for screening nonhuman primate populations for mycobacterial infections. Exposure to nontuberculous mycobacteria, such as M. leprae, amy sensitize some individual primates to nonspecific mycobacterial antigens, resulting in false-positive tuberculin reactions. Susceptibility of the cynomolgus macaque and other nonhuman primates to M. leprae should be re-evaluated. Cynomolgus macaques and, possibly, other nonhuman primates may serve as valuable experimental models of leprosy in humans. PMID- 9728447 TI - The occurrence of reactions and impairments in leprosy: experience in the leprosy control program of three provinces in northeastern Thailand, 1987-1995 [correction of 1978-1995]. I. Overview of the study. AB - AIM: This paper is the first in a series of three reports on the occurrence of reactions and impairments in leprosy in Thailand. This first paper gives a general overview about the methodology of the study, some epidemiological observations, delay in detection, multidrug therapy (MDT) completion rates and relapses. The other two papers report on: II. Reactions and III. Neural and Other Impairments. This study was carried out from 1987 until 1995 in three neighboring provinces in northeastern Thailand. STUDY DESIGN: A population-based, prospective cohort study. STUDY POPULATION: All 640 newly diagnosed leprosy patients in the three provinces, registered between October 1987 and September 1990, were included [420 paucibacillary (PB) and 220 multibacillary (MB)]. This group was followed up (actively and passively) until the end of 1995. METHODS: Patients were found by active and passive case finding. All new, untreated leprosy patients from the area were enrolled and started on the World Health Organization (WHO) MDT (WHO/MDT) regimen. A vertical control service was run by specialized leprosy workers. During treatment the patients received their monthly doses at home. During surveillance the patients were followed up once a year by a special team. Patients were questioned about delay in detection. Treatment completion rates were calculated. The occurrence of reactions and neural and other impairments at the beginning of, during and after treatment was ascertained. After treatment, the occurrence of late reactions and relapses was recorded. RESULTS: A higher frequency of leprosy was found among the male patients, especially in the MB group. However, in the PB group a higher female/male ratio was found in the age group 55 years and older. There was an increase in the detection rate from the youngest age group to the age group 55 years and older, which showed the highest detection rate. Treatment completion rates were high, 95% in both in the PB and MB treatment groups. About 50% of the new cases reported a delay between onset and registration of 1 year or more. By 1995, 93% of the original patient group was still available for follow up. By the end of 1995, 8 PB and 2 MB relapses were recorded. PMID- 9728448 TI - The occurrence of reactions and impairments in leprosy: experience in the leprosy control program of three provinces in northeastern Thailand, 1987-1995 [correction of 1978-1995]. II. Reactions. AB - AIM: This is the second paper in a series of three papers on the occurrence of reactions and impairments in leprosy in Thailand, and focuses on the prevalence and incidence of reactions, including silent neuropathy. STUDY DESIGN: A population-based, prospective cohort study. STUDY POPULATION: All 640 newly diagnosed and registered leprosy patients in three neighboring provinces in northeastern Thailand registered between October 1987 and September 1990 were included [420 paucibacillary (PB) and 220 multibacillary (MB)]. This group was followed up (actively and passively) until the end of 1995. METHODS: Clinical data and data on the sensibility and motor function of the eyes, hands and feet were obtained when appropriate. The occurrence of reactions, including silent neuropathy, at the beginning of, during and after treatment was ascertained. During surveillance mild late reactions were also recorded. RESULTS: Severe reversal reactions (RR) at the start of and during treatment were seen in 2.6% [confidence interval (CI) 1.1-4.1] of the PB and 29% (CI 23-35) of the MB patients. In the PB group the majority (82%) of severe RR were found at the start of treatment. In the MB group 35% of the severe RR were found at the start of treatment and another 59% during the first year of treatment. It is shown that there is a statistically highly significant increasing proportion of patients with severe RR starting from tuberculoid and going toward borderline lepromatous. The incidence rate of severe RR during treatment was 1.4 (CI 0.46-4.5) per 100 person-years at risk (PYAR) for PB patients and 12 (CI 9.0-16) per 100 PYAR for MB patients. Late (mild and severe) RR were seen in 2.7% of the PB and 9% of the MB patients (35% of these reactions being considered severe). Late reactions were mainly seen in borderline tuberculoid (PB group) and in borderline lepromatous patients. Recent silent neuropathies were seen at first examination and during treatment in 1.4% of the pB and 4.1% of the MB patients. During surveillance only a few silent neuropathies were seen. If all severe RR, severe erythema nodosum leprosum and silent neuropathies at the start of, during and after treatment were added together, then 53% of the borderline lepromatous and 42% of the lepromatous patients had or developed one or another serious complication in need of steroid treatment. PMID- 9728449 TI - The occurrence of reactions and impairments in leprosy: experience in the leprosy control program of three provinces in northeastern Thailand, 1987-1995 [correction of 1978-1995]. III. Neural and other impairments. AB - AIM: This the third paper in a series of three papers on the occurrence of reactions and impairments in leprosy in Thailand, and focuses on the prevalence and incidence of neural and other impairments in leprosy. STUDY DESIGN: A population-based, prospective cohort study. STUDY SUBJECT: All 640 newly diagnosed and registered leprosy patients in three provinces of northeastern Thailand between October 1987 and September 1990 were included [420 paucibacillary (PB) and 220 multibacillary (MB)]. This group of patients was followed up until the end of 1995. METHODS: Clinical data; data on the sensibility and motor function of eyes, hands and feet, and data on wounds and bone loss were obtained where appropriate. The occurrence of neural and other impairments at first examination, during treatment and during surveillance was ascertained. RESULTS: The relationship between impairment prevalence (grade 2 of the combined PB and MB groups and grades 1 and 2 together of the combined PB and MB groups) and duration of disease (before diagnosis) was found to be statistically significant. Increased delay in detection led to increased problems of impairments. Too many patients still develop new/additional impairments while on treatment and thereafter. The incidence of nerve function impairment (NFI) among patients without impairments at first examination while on treatment was 1.7 [ 95% confidence interval (CI) 0.45-4.4] per 100 person-years at risk (PYAR) for the PB group and 12 (CI 8.4-17) per 100 PYAR for the MB group. Additionally, 2% of the PB and 11% of the MB patients who already had impairments at first examination developed new NFI while on treatment. The outcome, comparing the first examination with the last examination during/after surveillance [changes in the voluntary muscle test (VMT), the sensory test (ST), wound count and bone loss], indicated that of the PB patients 3.7% improved, 3.7% got worse and 3.9% kept the same impairment; of the MB patients 19% improved, 18% got worse and 2.9% kept the same impairment. During treatment most of the new/additional impairments were due to new/increase in NFI; during surveillance slightly more than 50% were due to new/increase in NFI. Eighty-three percent of the MB patients without impairments at first examination who developed NFI during treatment improved (completely or partially) after receiving prednisolone. Only 62% of the MB patients with a grade 1 impairment at first examination and who developed a severe reaction or recent silent neuropathy improved after receiving prednisolone. There is a need for an indicator to measure new/additional impairments while on treatment and thereafter. It is proposed to measure changes in impairment by measuring changes in VMT, ST, wound count and bone loss. PMID- 9728450 TI - Why relapse occurs in PB leprosy patients after adequate MDT despite they are Mitsuda reactive: lessons form Convit's experiment on bacteria-clearing capacity of lepromin-induced granuloma. AB - It is amazing how after years of scientific research and therapeutic progress many simple and basic questions about protective immunity against Mycobacterium leprae remain unanswered. Although the World Health Organization (WHO) has recommended short-term multidrug therapy (WHO/MDT) for the treatment of paucibacillary (PB) leprosy patients, from time to time several workers from different parts of the globe have reported inadequate clinical responses in a few tuberculoid and indeterminate leprosy patients following adequate WHO/MDT despite the fact that they are Mitsuda responsive. A few borderline tuberculoid patients harbor acid-fast bacilli (AFB) in their nerves for many years even though they become clinically inactive following MDT, a fact which has been ignored by many leprosy field workers. Keeping these patients in mind, we have attempted to investigate the cause of the persistence of AFB in PB cases and have looked into the question of why Mitsuda positivity in tuberculoid and indeterminate leprosy patients, as well as in healthy contacts, is not invariably a guarantee for protectivity against the leprosy bacilli. We have: a) analyzed the histological features of lepromin-induced granulomas, b) studied the bacteria-clearing capacity of the macrophages within such granulomas, and c) studied the in vitro leukocyte migration inhibition factor released by the blood leukocytes of these subjects when M. leprae sonicates have been used as an elicitor. The results of these three tests in the three groups of subjects have been compared and led us to conclude that the bacteria-clearing capacity of the macrophages within lepromin-induced granuloma (positive CCB test) may be taken as an indicator of the capability of elimination of leprosy bacilli and protective immunity against the disease. This important macrophage function is not invariably present in all tuberculoid and indeterminate leprosy patients or in all contacts even though they are Mitsuda responsive and are able to show a positive leukocyte migration inhibition (LMI) test. It is likely but not certain that this deficit of the macrophage is genetically predetermined and persists after completion of short term WHO/MDT. Thus, after discontinuation of treatment slow-growing, persisting M. leprae multiply within macrophages leading to relapse. PMID- 9728451 TI - Serial Mitsuda tests for identification of reactional tuberculoid and reactional borderline leprosy forms. AB - The authors studied the Mitsuda reaction in 37 leprosy patients (18 reactional tuberculoid, 19 reactional borderline cases) and compared the results with clinical findings, histopathology and bacilloscopy. Evaluation of the Mitsuda reaction was carried out on days 30, 60, 90 and 120. Most of the reactional tuberculoid patients showed a Mitsuda reaction of +3 in opposition to the reactional borderline patients who showed only +. Bacilloscopic analysis revealed that in 75% of the reactional tuberculoid cases there were rare or no bacilli; bacilli were present in 95% of the reactional borderline cases. The authors conclude that reactional tuberculoid cases have a greater ability to clear bacilli than reactional borderline cases, and that the Mitsuda reaction is a useful tool for the differentiation between these two types of leprosy. PMID- 9728452 TI - Computed tomographic study of paranasal sinuses in lepromatous leprosy. AB - Twenty patients (18 males, 2 females) with lepromatous leprosy and 10 age- and sex-matched controls were subjected to computed tomographic (CT) scans of the paranasal sinuses (PNS). The mean bacterial (BI) and morphological indexes were 3.4+ and 1%, respectively. Nasal symptoms were present in nine (45%) patients; 15 (75%) out of 20 patients showed significant abnormalities on CT scans of the PNS compared to none of the 10 controls. The maxillary and ethmoid sinuses were affected in 11 (55%) patients each, followed by the sphenoid sinus in four (20%) patients. Frontal sinus involvement was least frequent, only one (5%) patient showed CT changes. Mucosal thickening was the most common finding followed by soft-tissue densities and, rarely, a fluid level was seen in the PNS. Involvement of the PNS correlated with a high BI of 4+ or more (75% vs 37.5%). Paranasal sinus involvement is an integral part of lepromatous leprosy since histological involvement was present even when the CT scan was apparently normal. PMID- 9728453 TI - Two-dimensional electrophoretic analysis of humoral responses to culture filtrate of Mycobacterium bovis BCG in patients with leprosy and tuberculosis. AB - Sera from 3 lepromatous (LL), 3 borderline lepromatous (BL), 3 mid-borderline (BB), 3 borderline tuberculoid (BT), 2 tuberculoid (TT) and 4 tuberculosis (TB) patients and 3 healthy individuals were examined for their reactivities against the proteins in the culture filtrate of BCG separated by two-dimensional electrophoresis (2-DE). The sera were obtained from patients who were untreated. Sera from LL and BL patients reacted strongly with the antigen 85 (Ag85) complex and MPB51. Sera from LL and BL patients also weakly reacted with the newly identified 29-, 24- and 23-kDa spots. Sera from the other patients reacted similarly, but the levels of reaction were remarkably lower than those form LL and BL patients. Mycobacterium leprae antigens that are analogous to BCG AG85 and MPB51 are suggested as the main targets for the humoral immunity of untreated patients. The reactivities of sera with newly identified antigens may provide the potential for predicting the severity and prognosis of diseases. PMID- 9728454 TI - Correlation of oral surface temperatures and the lesions of leprosy. AB - A short review of the literature on the optimum temperature for the growth of Mycobacterium leprae is followed by a report of an investigation into the correlation of oral surface temperatures with oral leprous lesions. It is concluded that the oral lesions of leprosy occur more frequently in areas of the mouth with a lower surface temperature. PMID- 9728455 TI - Toward the elimination of leprosy: the challenges and opportunities. PMID- 9728456 TI - The dangers of misinterpretation of the elimination campaign. PMID- 9728458 TI - Peptic ulcer and corticosteroid therapy. PMID- 9728457 TI - MDT and transmission. PMID- 9728459 TI - Leprosy and HIV infection in Bahia, Brazil. PMID- 9728460 TI - Arsenic poisoning mimicking polyneuritic leprosy. PMID- 9728461 TI - Giant lesions in histoid leprosy. PMID- 9728462 TI - Serologic assays using the 18-kDa antigen of Mycobacterium leprae expressed in the yeast Saccharomyces cerevisiae. PMID- 9728463 TI - Collagen profile in sciatic nerves of M. leprae-inoculated mice correlates with in vitro collagen production by Schwann cells. PMID- 9728464 TI - Triple pelvic osteotomy. PMID- 9728465 TI - Pheochromocytoma in the vertebral canal of two dogs. AB - Pheochromocytomas are uncommon tumors arising from the adrenal gland, which have potential for aggressive local spread. The diagnosis is extremely challenging, particularly when classical endocrine signs are absent. This paper presents two canine cases of pheochromocytoma in the vertebral canal and illustrates the potential for detection of the tumor by abdominal ultrasonography. PMID- 9728466 TI - Extradural spinal angiolipoma associated with bone lysis in a dog. AB - An extradural spinal tumor was diagnosed in a 12-year-old Labrador retriever that was presented with a one-week history of paraparesis. Myelography indicated a deviation of the spinal cord to the right side at the level of the second lumbar (L2) vertebra. The difference in length measuring the left and right sides of the L2 vertebra suggested a fracture of the vertebral body. Severe bone remodeling and an extradural mass were seen on computed tomography (CT). Clinical, radiographical, and histological findings are described and considered homologous to extradural angiolipomas described in the human literature. PMID- 9728467 TI - Realignment of a seventh lumbar vertebral fracture/luxation using a Senn retractor in two puppies. AB - Two puppies, each with fractures of the seventh lumbar (L7) vertebra, had their vertebral canals aligned surgically with the aid of a Senn retractor. The retractor was used to provide leverage during vertebral manipulations. Surgical fixation was achieved using Steinmann pins and polymethylmethacrylate. Both puppies were improved immediately after surgery and were clinically normal six months after surgery. PMID- 9728468 TI - Surgical correction of cor triatriatum sinister in a kitten. AB - A seven-month-old, male domestic shorthair was presented for respiratory distress. Cor triatriatum sinister was diagnosed based on echocardiography. Surgical dilatation and tearing of the anomalous membrane dividing the left atrium resulted in resolution of clinical signs. Intraoperative echocardiography was used to evaluate the adequacy of the repair. PMID- 9728469 TI - Construction of a weight-bearing surface on a dog's distal pelvic limb. AB - A one-year-old, neutered female boxer presented with a self-inflicted pandigital amputation following complications of a left hind footpad laceration repair. A meshed skin graft was placed distally over the exposed granulation tissue of the affected limb. In two surgical procedures, a total of five 6 by 8-mm and three 8 by 10-mm digital pad grafts were transplanted into recessed sites in the granulation tissue over the distal aspect of the metatarsal bones. A newly designed pressure relief bandage/ splint was used to assist maturation of the grafts. The result was a weight-bearing surface over an area of maximum tissue stress. PMID- 9728470 TI - Use of a Foley catheter to facilitate anal sac removal in the dog. AB - Chronic anal sac infection or impaction can be a frustrating problem for both the dog owner and veterinarian. Anal sacculectomy may be used to resolve clinical signs when medical management fails. Four dogs, ranging in size from 6 kg to 34 kg, were treated with closed anal sacculectomies, in which the balloon of a Foley catheter was used to facilitate surgical dissection of the sac. In all four cases, the Foley catheter successfully distended the anal sac during its removal. Clinical signs associated with the diseased anal sac were abated in the four dogs for a follow-up period of one-to-three years. PMID- 9728471 TI - Reconstruction of a facial defect using the ear pinna as a composite flap. AB - A four-month-old, female pit bull was evaluated for multiple, resolving, second- and third-degree burn wounds. The convex surface of the left pinna was severely burned and distorted. Contraction of a large (6 cm by 4 cm), facial cutaneous defect had resulted in contracture of surrounding normal skin and dorsal displacement of both upper eyelids. Decreased ability to blink predisposed the puppy to exposure keratitis. The cutaneous facial defect was repaired using the left pinna as a composite skin flap. Surgery resulted in a successful repair of the defect and immediate return of normal eyelid function. PMID- 9728472 TI - Comparison of anesthetic and cardiorespiratory effects of diazepam-butorphanol ketamine, acepromazine-butorphanol-ketamine, and xylazine-butorphanol-ketamine in ferrets. AB - Ten ferrets were used in a crossover study to determine the sedative effects of intramuscularly administered diazepam (3 mg/kg body weight)-butorphanol (0.2 mg/kg body weight)-ketamine (15 mg/kg body weight); acepromazine (0.1 mg/kg body weight)-butorphanol (0.2 mg/kg body weight)-ketamine (15 mg/kg body weight); and xylazine (2 mg/kg body weight)-butorphanol (0.2 mg/kg body weight)-ketamine (15 mg/kg body weight). All of the ferrets became laterally recumbent following the administration of each drug combination. The xylazine-butorphanol-ketamine combination induced significantly longer (p less than 0.05) durations of tail clamp analgesia (mean+/-standard deviation [SD], 81.0+/-19.1 min versus 20.5+/ 25.4 min and 30.0+/-26.9 min), dorsal recumbency (mean+/-SD, 94.6+/-13.6 min versus 75. 6+/-34.7 min and 55.2+24.8 min), and muscle relaxation suitable for endotracheal intubation (mean+/-SD, 67.1+/-23.0 min versus 7.0+/-22.1 min and 9.5+/-15.4 min) than the diazepam-butorphanol-ketamine and acepromazine butorphanol-ketamine combinations, respectively. The recovery time from dorsal recumbency to standing was not significantly different among the three treatment groups. The heart rate was significantly lower in the xylazine-butorphanol ketamine group; however, systolic blood pressure was not significantly different among the treatment groups. Ventilatory function was more depressed in the diazepam-butorphanol-ketamine and xylazine-butorphanol-ketamine groups than in the acepromazine-butorphanol-ketamine group. A period (approximately 45 minutes) of hypoxia was observed in the xylazine-butorphanol-ketamine-treated ferret. Of the three combinations evaluated in ferrets, xylazine-butorphanol-ketamine was concluded to be the most effective anesthetic combination. However, hypoxemia and ventricular arrhythmias were observed in the xylazine-butorphanol-ketamine treated ferrets, so the effectiveness of the xylazine-butorphanol-ketamine combination should be weighed against its cardiorespiratory side effects. PMID- 9728473 TI - Prognosis for dogs with stage I or II splenic hemangiosarcoma treated by splenectomy alone: 32 cases (1991-1993). AB - A retrospective analysis was performed on the case records of 32 dogs with Stage I or II splenic hemangiosarcoma that were treated by splenectomy alone and that survived the seven-day postoperative period. Median survival time for these 32 cases was 86 days (mean, 116 days; range, 14 to 470 days), and the one-year survival rate was estimated to be 6.25%. Survival was not influenced by signalment, presenting signs, stage of disease, or clinicopathological findings. The data provides a basis from which to evaluate adjuvant chemotherapy for splenic hemangiosarcoma that is confined to the spleen macroscopically. PMID- 9728474 TI - Prospective comparison of four sampling methods (cystocentesis, bladder mucosal swab, bladder mucosal biopsy, and urolith culture) to identify urinary tract infections in dogs with urolithiasis. AB - A prospective study was conducted on 27 dogs with recurrent urinary tract infections (UTIs) and urolithiasis. Four sampling methods (i.e., urine obtained by cystocentesis, bladder mucosal swab, bladder mucosal biopsy, and urolith) were compared to identify UTI. Identical culture results were obtained from urine collected by cystocentesis and from the swab of bladder mucosa. In the presence of a positive urine culture, the same organism also was cultured from the bladder mucosal biopsy and urolith. However, in the presence of a negative urine culture, an organism was cultured from the bladder mucosal biopsy or the urolith in 18.5% of the cases. Therefore, when the culture from urine obtained by cystocentesis is negative, it is recommended that aerobic cultures of a bladder mucosal biopsy and a urolith be performed in cases of urolithiasis. PMID- 9728475 TI - Mesenteric vein thrombosis in a dog. AB - A four-year-old, male cocker spaniel was presented for vomiting and anorexia of two days' duration. An elongated abdominal mass was palpated, and abdominal pain was noted. On exploratory laparotomy, a jejunal segment was found to be infarcted transmurally. Histopathology confirmed the diagnosis of mesenteric vein thrombosis. The dog recovered uneventfully following resection of the affected bowel. PMID- 9728476 TI - Hematometra secondary to anticoagulant rodenticide toxicity. AB - An adult, intact female Australian shepherd presented for frank vaginal bleeding of unknown duration. The only coagulation profile abnormality upon presentation was mild prolongation of the partial thromboplastin time (PTT). The uterus was removed at surgery and contained a large amount of coagulated blood. Clotting profiles were markedly abnormal 48 hours postoperatively. Serum analysis was positive for brodifacoum, an anticoagulant rodenticide. Preoperative coagulation was most likely normalized by vitamin K1 therapy administered prior to presentation. The only manifestation of anticoagulant rodenticide was hematometra. Rodenticide intoxication should be considered in the differential diagnosis list of hematometra or metrorrhagia. PMID- 9728477 TI - Role of sarcoplasmic reticulum in the contractile dysfunction during myocardial ischaemia and reperfusion. AB - In the myocardium, the sarcoplasmic reticulum (SR) plays an essential role in the regulation of cytosolic free Ca2+ ion concentration and, hence, in the contraction-relaxation cycle. The aim of this review is to summarize the role of the SR, particularly the main SR Ca2+ transport proteins, Ca2+-ATPase pump and Ca2+ release channel (ryanodine receptor), in contractile impairment during ischaemia and reperfusion. As suggested by most studies, SR dysfunction may contribute to contractile failure during ischaemia. However, SR function is largely restored during reperfusion and minor changes are unlikely to explain the severe postischaemic contractile dysfunction. PMID- 9728478 TI - Pulmonary haemodynamics in acute experimental lung vascular injury. AB - Acute lung injury was induced by intravenous injection of 20 microl of a mixture of equivalent volumes of capronic acid, caprilic acid and olive oil in intact anaesthetized rats and in isolated perfused rat lung preparations. Lung injury in intact rats resulted in an increase in lung weight related to body weight and in a decrease in the lung dry/wet weight ratio. Lung compliance, measured in a body plethysmograph, was decreased. PaO2 decreased and PaCO2 increased in 10 and 20 min, respectively, after the beginning of the experiment. Mean blood pressure in pulmonary artery increased immediately after the injection. Isolated rat lungs were perfused at constant flow with physiological saline solution containing bovine albumin and meclofenamate. The injection of a mixture of capronic acid, caprilic acid and olive oil increased the baseline perfusion pressure and led to a release of endothelial cells into the perfusate. The perfusion flow-pressure relationship was shifted upwards. Both the extrapolated pressure axis, intercept and slope of the plot were significantly elevated. The described experimental lung injury is a suitable model for studies on the effects of vascular wall damage and transvascular fluid leak in pulmonary vasculature. PMID- 9728479 TI - 2'(3')-O-[N- [2- [3- [5-fluoresceinyl] thioureido] ethyl] carbamoyl] adenosine 5' triphosphate and its Cr(H2O)4 and Co(NH3)4 complex derivatives are new fluorescent tools for labelling ATP binding sites of Na+/K+-ATPase. AB - 2'(3')-O-[N- [2- [3- [5-fluoresceinyl] thioureido] ethyl] carbamoyl] adenosine 5' triphosphate (FEDA-ATP), a spectroscopic tool used for studying skeletal muscle myosin ATPase subfragment 1, was applied to Na+/K+-ATPase (EC 3.6.1.37). In contrast to the myosin subfragment, we found that FEDA-ATP is not a substrate for Na+/K+-ATPase. On the other hand, FEDA-ATP showed an affinity for both the low (E2, Kd=200 microM) and the high (E1, Kd=22 microM) affinity ATP-binding sites. When the microscopic affinities of FEDA-ATP were used for calculating the macroscopic affinity in the overall reaction according to Ki=(KdE1*KdE2)1/2, the experimentally measured inhibition constant of 66 microM was obtained. To evoke irreversible binding inhibitors, FEDA-ATP was transferred in its chromium(III) and cobalt(III) complex analogs, which are suitable tools for labelling the ATP binding sites of Na+/K+-ATPase in a specific way. PMID- 9728481 TI - Molecular variants of the renin-angiotensin system components in the Slovak population. AB - Molecular variants of individual components of the renin-angiotensin system (RAS) are reported to constitute the inherited predisposition to some cardiovascular diseases in man, e.g. essential hypertension or myocardial infarction. The frequency of these variants depends highly on the race and population. Therefore, we examined the M235T molecular variant of the angiotensinogen gene and the I/D polymorphism of the ACE gene in Slovak healthy population, in patients with diagnosed essential hypertension and in patients who had undergone myocardial infarction. DNA from 241 subjects was tested for the presence of M235T and I/D molecular variants. The frequency of both these polymorphisms in the Slovak population is similar to other Caucasian populations. In the group of hypertensive patients, the frequency of the M235T molecular variant was increased compared to controls, predominantly in males (0.45 vs. 0.28), while in the I/D polymorphism the incidence of the D allele was the same for both controls and hypertensives (0.49 vs. 0.50). A significant increase in the D allele frequency compared to the controls occurred in the group of infarcted patients (0.63). The increased frequency of the M235T allele in hypertensive patients compared to the healthy population confirms that the M235T variants associated with increased blood pressure in the Slovak population. In the Slovak population, I/D polymorphism of the ACE gene is associated with myocardial infarction rather than with hypertension. PMID- 9728480 TI - The localisation of TPPS4 in some organs and its possible nephrotoxicity in rats. AB - Photodynamic therapy (PDT) is now being used more frequently in carefully selected cases of malignancies. The drugs used for PDT are mostly derivatives of haematoporphyrine (HPD) and its active component photofrine II. Another compound prepared by total synthesis is meso-tetra-(4-sulfonatophenyl)-porphine (TPPS4) but its application in human medicine was rejected because of its neurotoxicity. Our TPPS4 was prepared by the method of Busby et al. in the modification of Jirsa and Kakac (1987). This product is purer and without neurotoxic effects. In this study, we concentrated our attention on the effect of TPPS4 on nephrotoxicity and its accumulation in some organs. As the parameters of toxic kidney damage we used urine levels of N-acetyl-beta-D-glucosaminidase (NAG), serum creatinine levels, glomerular filtration rate (GFR) and proteinuria. TPPS4 was administered i.v. in a dose of 25 mg/kg b.w. The animals were observed for 21 days after drug application. Urine and blood samples were collected over 24-hour periods on days 0, 5 and 21. The serum creatinine level was significantly higher only on day 5 (65.0+/-1.46 micromol/l vs 56.5+/-2.69 micromol/l on day 0, p<0.05). There were no significant changes in GFR, proteinuria or NAG activity in the urine during the experiment. AST serum activity was increased. We determined the concentration of TPPS4 (pmol/mg w.w.) in rat organs on the 21st day after the injection. The concentration of TPPS4 was high in kidneys (30.8+/-5.5), liver (13.5+/-2.0), lungs (11.7+/-4.6) and spleen (9.7+/-1.5), while the concentration in heart and brain was low. We conclude that TPPS4 has the highest concentration in the kidney 21 days after its administration and does not exert any nephrotoxic effects during this period. PMID- 9728482 TI - Effects of nitroprusside as a nitric oxide donor on anoxia/reoxygenation and D galactosamine hepatic injuries: a study in perfused hepatocytes. AB - At present, the physiological role of NO. synthesis in the liver is ambiguous. Studies directed to reveal the role of NO. in relation to liver function were primarily initiated by an interest in the hepatic response to infections and the consequent modulation of liver function. The purpose of the present investigation was to use perfused rat hepatocytes to test the ability of the latter to produce NO. and to delineate the relationship between exogenously delivered NO. and any alteration in the degree of injury as produced by anoxia/reoxygenation (AR) or D galactosamine (GalN, 5 mM) intoxication. NO. production in rats was stimulated by a single dose of lipopolysaccharide (LPS, 20 mg/kg i.p.) from which hepatocytes were isolated and perfused. Exogenous NO. was delivered to the perfusate of hepatocytes that were isolated from untreated rats, by the addition of sodium nitroprusside (SNP, 2 mM and 0.2 mM). AR and GalN hepatocyte injury was followed after the addition of SNP. Rat hepatocytes were immobilized in low-gelling agarose and perfused with Williams E medium. Endogenous synthesis of NO. and exogenous NO. as produced by SNP was evaluated by estimating the end products of NO. (NO2- + NO3-) in the perfusion medium. The functional and structural integrity of hepatocytes was evaluated from lactate dehydrogenase (LD) leakage and urea synthesis in the perfusion medium. Normal, AR- and GalN-injured hepatocytes did not exhibit measurable NO. while LPS-treated hepatocytes produced NO. (80 microM NO2- + NO3-). SNP-produced NO. significantly increased or decreased LD leakage in AR at 2 mM or 0.2 mM, respectively, and also reduced or increased the rate of urea synthesis, respectively. 0.2 mM SNP increased trypan blue exclusion by hepatocytes. On the other hand, GalN toxicity was not significantly altered by SNP as demonstrated by LD leakage and the rate of urea synthesis was increased by SNP addition. The present data suggest both deleterious and beneficial role of NO. in AR liver injury model depending on the level of NO. generated. PMID- 9728483 TI - An in vitro study on methotrexate hydroxylation in rat and human liver. AB - Methotrexate (MTX) was investigated for possible effect on the metabolism of ethoxyresorufin, pentoxyresorufin and ethoxycoumarin, the model substrates of cytochrome P450. The investigation was carried out in liver microsomes of rats pretreated with classical inducers of cytochrome P450 as well as in microsomes of two human livers. Furthermore, we measured the conversion of MTX (100microM) to its main metabolite, 7-hydroxymethotrexate (7-OHMTX), in microsomes and cytosolic fractions of rat and human livers. The inhibition of 7-OHMTX formation by menadion (inhibitor of aldehyde oxidase) and allopurinol (inhibitor of xanthine oxidase) was studied in the cytosol of rat and human livers. In both species, MTX in the concentration range 0.5-500 microM exerted no inhibitory effect on enzymatic activities associated with cytochrome P450. Moreover, we did not observe any measurable formation of 7-OHMTX in liver microsomes. MTX was metabolized at a similar rate in the cytosol of rat and human liver. Allopurinol (100 microM) reduced the rate of MTX hydroxylation by 31.5% in the cytosol of human livers but had no effect in the rat. Menadion (100 microM) decreased the rate of 7-OHMTX formation in the cytosol of human and rat liver by 69% and 94%, respectively. Our results confirmed that MTX is oxidized by a soluble enzymatic system in both the rat and human liver. In human tissues, both aldehyde oxidase and xanthine oxidase may play an important role in the metabolism of MTX. Depression of cytochrome P450 and related enzymatic activities observed in vivo cannot be explained by a direct inhibitory action of MTX on cytochrome P450. PMID- 9728485 TI - Biological half-life of bromine in the rat thyroid. AB - The biological half-life of bromine in the rat thyroid was determined by measuring the radioactivity of thyroids of animals which continuously received 82Br labelled bromide in their food. The value of this half-life (110 h) is practically the same as the biological half-life of iodine. The rate of establishing the I/Br concentration ratio in the thyroid depends on the biological half-life of bromine. The mechanism of this process depends on the state of iodine supply. When the supply is sufficient, the iodine concentration in the thyroid remains constant, while during iodine deficiency the iodine atoms are replaced by atoms of bromine. PMID- 9728484 TI - Superoxide dismutase activities in different tissues of female rats treated with olive oil. AB - The activities of cytosol superoxide dismutase (CuZnSOD) and mitochondrial superoxide dismutase (MnSOD) were measured in subcellular fractions of homogenates prepared from the brain, thymus and liver of ovariectomized (OVX) female rats, non-treated or treated 24 h prior to sacrifice with a single s.c. dose of 0.1 ml olive oil. In the brain, neither MnSOD nor CuZnSOD were affected by olive oil, whereas in the thymus the olive oil injection elevated CuZnSOD and did not affect MnSOD activity. At the same time, the activity of CuZnSOD was reduced and that of MnSOD was elevated in the liver following oil treatment. These results suggest that olive oil has modulatory effects on the expression of CuZnSOD and MnSOD activity in the liver and of CuZnSOD in the thymus of female rats. PMID- 9728486 TI - Activities of superoxide dismutase and catalase in erythrocytes and transaminases in the plasma of carps (Cyprinus carpio L.) exposed to cadmium. AB - Total superoxide dismutase (SOD, EC 1.15.1.1) and catalase (CAT, EC 1.11.1.6) activities in erythrocytes and the glutamic acid-oxalacetic acid-transaminase (GOT, EC 2.6.1.1) and glutamic acid-pyruvic acid-transaminase (GPT, EC 2.6.1.2) activities in the plasma were measured in experimental groups of carps (Cyprinus carpio L.) exposed to cadmium in a concentration of 20 mg Cd/l water under aquarium conditions for 6, 12, 18 and 24 hours and in control fishes. It was shown that the total activity of SOD in the erythrocytes is significantly decreased after 12, 18 and 24 hours of cadmium exposure. Increased activities of CAT (after 24 hours) in the erythrocytes and GOT and GPT in the plasma were found in cadmium-treated fishes. At the same time the concentration of blood haemoglobin and haematocrit values were significantly diminished. These results indicate that cadmium causes oxidative stress and tissue damage in the exposed fishes. PMID- 9728487 TI - Two models of epileptic spike-and-wave rhythm in rats are differently influenced by ethosuximide. AB - Epileptic afterdischarges induced by electrical stimulation of the sensorimotor cortex as well as minimal metrazol seizures are characterized by EEG spike-and wave rhythm and nearly the same motor pattern of clonic seizures. The action of ethosuximide on these two models was tested in adult rats with implanted electrodes. Cortical afterdischarges remained practically uninfluenced by ethosuximide (62.5 or 125 mg/kg i.p.) whereas minimal metrazol seizures were suppressed in a dose-dependent manner (doses of 31.25, 62.5 and 125 mg/kg i.p. were used). Present results in connection with recent data on the abolition of spike-and-wave rhythm elicited by low systemic doses of pentylenetetrazol suggest that spike-and-wave rhythm does not represent a single entity. PMID- 9728488 TI - Sex-dependent differences in growth and morphology of cultured vascular smooth muscle cells from newborn rats. AB - The morphology and proliferation of vascular smooth muscle cells (VSMC) were studied in cultures prepared from the aorta of newborn male and female Wistar rats. The doubling times (DT) of the male-derived population were 16.4+/-0.7 h and 30.0+/-2.2 h in the exponential and post-exponential growth phases, respectively. In the female donor cells, the corresponding DT values were significantly longer, i.e. 21.9+/-1.8 h and 38.0+/-2.2 h. In addition, the period of growth was shorter in the female-derived cultures. The percentage of 3H thymidine labelled cells in male cultures was 61.0+/-3.1, 92.8+/-1.9 and 98.7+/ 0.6% at 2, 27 and 52 h, respectively. In the female-derived populations, only 24.6+/-4.4, 66.1+/-3.8 and 82.8+/-2.0% of cells were labelled at the corresponding incubation intervals. As a consequence, the final population density in male cultures was 5.6 times higher. In addition, the male-derived VSMC were mainly spindle-shaped and bulgy in appearance while those from female donors were flat and polygonal which means that the cells were adhering to the growth support to a different extent. The study revealed early determination and long term persistence of lower adhesiveness as well as higher growth potential of male VSMC, i.e. properties which may be of importance for explaining the higher incidence of vascular wall disorders in males. PMID- 9728489 TI - Distribution of nitric oxide in cardiovascular system. AB - We report here the in vitro measurements of nitric oxide in the cardiovascular system using a porphyrinic sensor specific for NO. Nitric oxide concentrations were measured directly in different parts of the heart and also in different arteries and veins, ranging from 100 microm to 5 mm in diameter. Highest NO. concentrations were found in the heart and particularly in the areas of aortic and pulmonary valves. The NO. concentration in the arteries was higher than in the veins. A clearcut positive correlation was obtained by plotting the vessel diameter and production of nitric oxide. PMID- 9728490 TI - New strategy for prolonging the preservation time of hearts for transplantation. AB - Our study concerned the findings that rat and rabbit heart transplants do not survive after six hours. They become dark, hard and fail to contract within 2 min after reperfusion and never regain their function. We tested the supplementation of solutions for heart transplant preservation with tetrahydrobiopterin (H4B) and L-arginine (L-ARG) to maintain the oxidative and reductive domains of the endocardial NO synthase. We decided to study the excised rabbit hearts preserved in Hank's balanced salt solution (HBSS) at 0 degrees C supplemented with different concentrations of H4B (0, 1, 5, 10 or 100 microM). At desired time intervals, successive pieces stored in the above solutions were warmed to rabbit body temperature in 4 ml of HBSS and maximally agonized by direct application of 20 microl of 200 microM bradykinin (or other agonist) onto the exposed endocardium. Nitric oxide bursts were monitored with a porphyrinic NO sensor lying on the exposed endocardium. Our goal was to find the lowest H4B concentration which would maximally agonize NO. and prolong the time of heart preservation to more than 6 hours. Ten microM are a minimum H4B concentration which achieves maximum prolongation of heart preservation time up to 90 hours. This effect was based upon maximal potentiation of NO. release and minimizing of superoxide production. PMID- 9728491 TI - Relationships between transmembrane action potential changes and simultaneous changes in electrocardiograms of rats after a one-month aortic pressure overload. AB - An attempt was made to determine the relationship between the characteristics of electrical activity of the hypertrophied myocardium of rats at the cellular level and at the level of the whole heart after a one-month left ventricular pressure overload. Such an animal model has already been demonstrated to be highly resistant to epinephrine-induced arrhythmias. Since severe ventricular arrhythmias often occur in patients with cardiac hypertrophy, ventricular vulnerability might depend on some electrophysiological characteristics of the heart related to the stage of hypertrophy. Using the "floating" microelectrode technique, the computed characteristics of cardiac transmembrane action potentials (AP) of the left and right epicardium cells were compared in situ to computed characteristics of the electrocardiograms in anaesthetized control rats (group C) and in rats with left ventricular hypertrophy (group H) induced by a one-month suprarenal constriction of the abdominal aorta. The aortic pressure overload caused a significant (p<0.001) and marked increase in AP duration of left ventricular cells: APD 30 and APD 80 were 29+/-3 ms and 89+/-6 ms, respectively, in group H and 14+/-1 ms and 53+/-2 ms in group C. The same modifications were observed in right ventricular cells when right hypertrophy was present. Simultaneous electrocardiograms exhibited a significant (p<0.01) prolongation of P-R, Q-S and T duration and T wave flattening in group H (63+/-2 ms, 32+/-3 ms, 109+/-5 ms and 0.25+/-0.03 mV as compared with 53+/-1 ms, 20+/-1 ms, 88+/-2 ms and 0.40+/-0.04 mV in group C). After a one-month aortic overload in rats, both left and right ventricles are hypertrophied and have the same electrophysiological characteristics: in this model, at this stage of hypertrophy, some factors favouring ventricular arrhythmias are missing. The corresponding flattening of the T wave in the ECG might be of clinical relevance. PMID- 9728492 TI - Changes in parameters of mechanics of breathing during high-frequency jet ventilation: what is the cause? AB - In experiments on 51 healthy anaesthetized and paralyzed rabbits the changes in parameters of mechanics of breathing during high frequency jet ventilation (HFJV) were determined and the mechanisms responsible for these changes were investigated. In the first series of experiments with two groups of animals ventilated by HFJV with relative inspiratory time ti=0.5 and ti=0.7 airway resistance (Raw) after 5 h of HFJV in the ti=0.5 group increased from 1.14+/-0.05 to 2.31+/-0.09 kPal(-1) x s (P < or = 0.001), in the ti=0.7 group from 1.22+/ 0.04 to 1.78+/-0.08 kPal(-1) x s (P < or = 0.01). Dynamic compliance (Cdyn) decreased in the ti=0.5 group from 0.041+/-0.004 to 0.017+/-0.001 l x kPa(-1) (P < or = 0.01) and in the ti=0.7 group from 0.034+/-0.003 to 0.022+/-0.002 l x kPa( 1) (P < or = 0.01). In the second series of experiments a group of animals was ventilated by HFJV after cervical vagotomy. The deterioration of Raw and Cdyn was significantly reduced in vagotomized rabbits in comparison to the controls without vagotomy. Finally, the study of phospholipid content in bronchoalveolar lavage fluid revealed no significant differences after 5 h of artificial ventilation or spontaneous breathing. These data indicate that HFJV results in changes in the parameters of mechanics of breathing in healthy lungs, which may be attenuated, but not fully eliminated, by bilateral cervical vagotomy. The decrease in Cdyn and increase in Raw are probably not due to changes in the pulmonary surfactant content. PMID- 9728493 TI - Kidney function changes in rats after single-dose administration of borocaptate sodium. AB - Kidney function changes after single-dose administration of borocaptate sodium (mercaptoundecahydro-closododecaborate, B12H11SH, BSH) were studied in rats. Changes of glomerular filtration rate (GFR) measured as 14C-inulin clearance, renal plasma flow rate (3H-p-aminohippuric acid clearance) and urine flow rate (UFR) after a slow intravenous injection of BSH (25 mg/kg b.w.) were investigated in rats under pentobarbital anaesthesia. It was found that a slow BSH injection induces a gradual decrease of renal plasma flow and glomerular filtration rate resulting in an almost constant reduction of the filtration fraction. These alterations were accompanied by a temporary increase of urine flow rate. Although a direct effect of BSH on the nephron cannot be excluded, it is suggested that the observed changes in kidney function might at least partly be mediated by disturbances in the function of the cardiovascular system following BSH injection. The role of the dianionic sulfhydryl group present in the borocaptate molecule in inducing these renal functional changes is discussed. PMID- 9728494 TI - Enhancement of haemopoietic spleen colony formation by drugs elevating extracellular adenosine: effects of repeated in vivo treatment. AB - The potential role of adenosine receptor signalling in the amplification of haemopoietic stem cells in vivo was investigated. Elevation of extracellular adenosine in mice was induced by the joint administration of dipyridamole, a drug inhibiting the cellular uptake of adenosine, and adenosine monophosphate, an adenosine prodrug. The response of haemopoietic stem cells to the drug treatment was measured by endogenous spleen colony-forming assay in sublethally gamma irradiated animals. The combination of drugs was administered before irradiation either singly or repeatedly at 24 h intervals. The results demonstrated the possibility of enhancing the spleen colony formation by the drug treatment. The highest stimulatory effect on spleen colony counts and on the colony sizes occurred after 3-4 injections of the drugs. Higher spleen colony responses were observed under injection regimens terminated 3 h before irradiation, as compared to those terminated 24 h before the radiation exposure. The results are interpreted as an evidence of the expansion of the stem cell pool. A tolerance to this stimulatory action developed after more than 3 injections of the drugs. PMID- 9728495 TI - The decrease of serum leptin levels in oestrogen-treated male mice. AB - Adipocyte hormone leptin (OB protein) is considered to be an "adiposity signal" regulating body weight homeostasis and energy balance. We have previously reported that oestrogens (oestradiol-benzoate) significantly decrease the body weight in male rats, increase anterior pituitary and serum levels of the intracellular messenger cAMP, which activates cAMP-dependent protein kinase A, their targets include hormone-sensitive lipase and they influence the brain sympathetic system. The present study tested our hypothesis that oestrogens could influence serum leptin levels in male mice. We found that chronic administration of oestradiol-benzoate significantly attenuated serum levels of leptin, in the dependence on the duration of its administration, and simultaneously decreased body weight. We suppose that oestrogens affect leptin levels interacting with the signal transmission system of cAMP, possibly at the genome level. Our observations that the food consumption of mice with simultaneously decreased body weight and levels of serum leptin support the idea that there exists a satiety factor that counters the effect of low leptin. PMID- 9728496 TI - Interstitial adenosine and serotonin in the feline lumbar spinal cord during short-term ischaemia. AB - During controlled ischaemia (aortal snare occlusion) of the lumbar spinal cord, microcirculatory (laser-Doppler flowmetry) and segmental neurophysiological parameters (monosynaptic reflexes, polysynaptic reflexes, cord dorsum potential=CDP) as well as interstitial concentrations of adenosine and serotonin (5-HT) were determined in the grey matter using the microdialysis/HPLC method. Ischaemic periods of 1-7 min with a residual blood flow in the lumbar spinal cord of 10-30 % of the preischaemic control blood flow caused a blockade of spinal pathways and an increase of concentrations of interstitial adenosine and 5-HT. This increase started immediately after the initiation of the ischaemic period and reached a maximum at the end or shortly after the end of the ischaemic period during postischaemic hyperaemia. A close correlation between the duration of ischaemia and the interstitial concentration of adenosine and 5-HT was not found. Repetition of ischaemic periods in an experiment did not lead to an extracellular accumulation or an exhaustion of the release of 5-HT, whereas some indication was found for an exhaustion of adenosine release. The course of the increase of interstitial adenosine and 5-HT was partly found to correlate to the loss and recovery of the CDP following ischaemia. The concentrations usually reached control levels before spinal reflexes reappeared. The highly dynamic changes in concentrations of adenosine and 5-HT in the extracellular space of the spinal cord during and after short-term ischaemia revealed some relation to the time course of recovery of segmental spinal functions by reflecting the course of spinal neuronal metabolism. PMID- 9728497 TI - Comparison of microsurgical suture with fibrin glue connection of the sciatic nerve in rabbits. AB - The regeneration of the sciatic nerve after microsuture was compared with the connection of transected nerve with a coagulum of autologous blood plasma in 20 rabbits. The epineuroperineural suture was performed in 10 rabbits (group A). The severed nerve was approximated with fibrin glue of autologous blood plasma in 10 rabbits (group B). Their skin sensation margin during a 3-month-period of regeneration was examined, 90 days after surgery the connection was inspected and the nerve conduction velocity was measured across the site of the anastomosis. The microsuture was found to be firm in all 10 animals of group A. On the other hand, in 2 animals of group B, the glue failed to keep the nerve stumps approximated (dehiscence occurred in 20% of the animals). There were no significant differences found on clinical and electrophysiological testing of regenerated nerves of both groups. The method of autologous fibrin glue in the repair of peripheral nerve transection does not provide a sufficiently firm connection. This procedure with the preparation of the centrifuged plasma is a more time-consuming method in comparison with the microsuture. Epineuroperineural microsuture with maximal effort to adapt the corresponding nerve fibres remains the method of choice for peripheral nerve reconstruction. PMID- 9728498 TI - The assessment proposal for long-term and short-term tolerable hygrothermal microclimatic conditions. AB - A group of four efficient mine rescuers 25 to 35 years old were exposed to a load of a cyclo-ergometer (stages A and B) and a hand ergometer (stage E) in a climate chamber. The total 120 min period of work was divided into four work intervals, 30 min each. There were 5-min breaks between the individual intervals. The load on the ergometer was selected in the range of 25 to 150 W, Tg=20 to 40 degrees C, rh=40 to 80% and v(a)=0.2 to 1.5 m x s(-1). The thermal resistance of the working suit was 0.65 clo at stage A, 1.07 clo at stage B and 0.81 clo at stage E. A total of 200 experiments with 50 combinations of the work and climate loads were made. The heart rate, oxygen consumption, carbon dioxide production, body temperature, skin temperature, water loss by sweating and perspiration, dry and wet bulb air temperature, air velocity and globe temperature were measured during the experiments. The expected production of sweat (SR) and the amount of accumulated heat in the body (Qmax) were calculated for each combination of the work-climate conditions by a computing program ISO 7933:1989 as well as by our own program. Good agreement was reached between the measured and predicted SR values, calculated by the ISO program (r=0.871) as well as between the values calculated by the two programs, respectively (r=0.985). The experimental results have shown good agreement between the predicted and actually measured values of temperature of the body core as an index of short-term tolerable climate load. The values of short-term tolerable time of work calculated at the level of accumulated heat in the body of 50 W x h x m(-2) resulted in an increase of body core temperature by 0.8 to 1.0 K. The values of heart rate did not mostly exceed 140 beats/min, reaching in exceptional (three) cases values above 150 beats/min. The authors recommend to limit the long-term work-heat (climatic) load during a higher metabolic rate (M >80 W x m(-2) including the basal metabolic rate) of acclimatized males and females at a sweat rate SR=270 g x h(-1) x m(-2), of non acclimatized persons at SR=206 g x h(-1) x m(-2). The limit for low metabolic rates (M < or = 80 W x m(-2)) for non-acclimatized and acclimatized persons is proposed for long-term tolerable loads of SR=147 g x h(-1) x m(-2). The short term tolerable load by heat storage within the organism for all categories is proposed as Qmax=50 W x h x m(-2). PMID- 9728499 TI - Surface tension of blood. AB - The surface tension of blood assessed in a group of 71 healthy subjects (24 men and 47 women) by the drop method at a temperature of 22 degrees C was 55.89 x 10( 3) N x m(-1), S.D.=3.57 x 10(-3) N x m(-1). It did not correlate with age or sex of the examined subjects nor with any of the following variables: red cell sedimentation rate, blood haemoglobin levels, number of erythrocytes, total serum cholesterol, total serum triacylglycerols, creatinine blood levels, ALT and AST activity. The surface tension of blood and other body fluids can play an important part not only in the genesis and development of decompression sickness but also in other processes in the organism. PMID- 9728500 TI - Circadian oscillations of lipid peroxides in the rat pineal gland. AB - We studied the circadian oscillation of lipid peroxides (TBARS) in the pineal gland of rats adapted to light:dark 12:12 h regimen. The concentration of TBARS was determined at 3-h intervals during 24 hours. TBARS of pineal gland oscillated rhythmically during the 24 h period. The maximal concentration of lipoperoxidative products was found at 20.00 h and 02.00 h and the lowest values at 08.00 h and 23.00 h. The determination of antioxidant capacity is needed for explaining the mechanism of TBARS oscillations in the pineal gland. PMID- 9728501 TI - Dose-dependent hypocholesterolaemic effect of oyster mushroom (Pleurotus ostreatus) in rats. AB - A highly significant negative correlation (r=-0.981, p < 0.001) between the amount of oyster mushroom (Pleurotus ostreatus) in the diet and cholesterol levels in the serum has been found in male Wistar rats fed shortly after weaning by a a diet with 0.3% cholesterol. The addition of 1.0, 2.5 and 5.0% of oyster mushroom to the diet reduced the levels of serum cholesterol by 11, 31 and 46%, respectively. The diet containing 5% of oyster mushroom suppressed cholesterol accumulation in the liver and increased the fraction of cholesterol carried by high-density lipoproteins. PMID- 9728503 TI - Effect of respiration and posture on heart rate variability. AB - The physiological control system of the heart produces a highly complex pattern of cardiac rhythmicity which is reflected in the variability of heart rate. The aim of this study was to analyse the effects of posture and breathing frequency on the cardiac control system by various noninvasive techniques. Seven healthy subjects (24+/-5 years, mean age+/-S.D.) were studied in the supine and sitting positions while breathing spontaneously or at a fixed rate (3, 6, 12, 24, 48, 60 breaths.min(-1)). Time series of instantaneous beat-to-beat heart rates were evaluated by spectral analysis and by the dimensionless approximate entropy parameter. The total spectral power as well as the low (<0.05 Hz) and mid frequency (0.05-0.12 Hz) spectral components were higher in the sitting position. Mean approximate entropy (+/-S.D.) (0.85+/-0.15 in sitting and 0.87+/-0.16 in lying subjects) was unaffected by postural changes or breathing frequencies higher than 6 breaths x min(-1). Analysis in the frequency domain revealed that the activity of the autonomic components controlling heart rate was modified by ventilation and postural changes, whereas approximate entropy, a unique measure of the complexity and integrity of the cardiac control system, was almost unaffected by respiration and posture. PMID- 9728502 TI - Treatment of neonatal rats with monosodium glutamate attenuates the cardiovascular reactivity to phenylephrine and angiotensin II. AB - In rats, neonatal administration of monosodium glutamate (MSG) causes serious damage in some hypothalamic and circumventricular areas. The resulting loss of appropriate neurons important for the regulation of blood pressure (BP) may modulate cardiovascular system receptivity in these animals. In the present study, the reactivity of the cardiovascular system to intravenous injection of alpha1-adrenergic receptor agonist phenylephrine (200 microg/kg/ml) and angiotensin II (500 ng/kg in 0.6 ml for 2 min) was investigated in adult rats which had been neonatally treated with MSG or vehicle. BP parameters measured directly in conscious cannulated rats were continuously registered using a computerized system. Under basal conditions, MSG-treated rats had slightly lower systolic, diastolic and mean BP with significant differences in pulse pressure (systolic - diastolic BP). In MSG-treated animals, the maximal increase of mean arterial BP after phenylephrine and the duration of BP elevation after both agents were significantly reduced. Slopes of the linear portion of baroreceptor function curves in control and MSG-treated rats did not differ significantly, indicating that baroreflex efficacy was unchanged. The results obtained by perfusion of the hindlimb vascular bed in situ showed that the pressure responses to increasing doses of noradrenaline in MSG-treated rats were reduced. These findings demonstrate that neonatal treatment of rats with MSG lowers the responsiveness of the cardiovascular system, particularly in response to alpha adrenergic stimulation. It is suggested that the attenuation of cardiovascular reactivity in MSG-treated rats is, at least partly, caused by diminished vascular responsiveness. PMID- 9728504 TI - Modification of organ protein synthesis after surgical stress by low energy diets with different supplements. AB - We have studied the effects of hypocaloric diets with different supplements on liver and jejunal mucosa protein synthesis. The supplements assayed were medium chain triglycerides (diet MCT, with 50% carbohydrates: 25% long chain triglycerides (LCT): 25% medium chain triglycerides (MCT), standard amino acids), branched-chain amino acids (diet BCA, identical to control diet L50, with 15.3% of nitrogen replaced by branched-chain amino acids) and glutamine (diet GLN, identical to diet L50, with 15.3% of nitrogen replaced by glutamine). The control diet (L50) had 50% carbohydrates: 50% LCT and standard amino acids. The diets were assayed on 86 rats with femoral fracture immobilized by Kirschner pin insertion. Nutrition was administered for 4 days. On the fifth day, liver and jejunal mucosa protein synthesis was determined. A branched-chain amino acid supply in a proportion higher than 21.2% of amino acid nitrogen significantly decreased liver and jejunal mucosa protein synthesis, while the same amount of glutamine did not modify it. MCT had no effect on jejunal mucosa protein synthesis, while it was decreased significantly in the liver. PMID- 9728505 TI - Modification of organ protein synthesis after surgical stress by low energy diets with different lipid/glucose ratios. AB - The aim of this work was to study the effects of low energy parenteral diets with different lipid/glucose ratios on rat liver and jejunal mucosa protein synthesis. The studied diets were: L0 (100% glucose, control diet), L25 (25% lipids: 75% glucose), L50 (50% lipids: 50% glucose) and L75 (75% lipids: 25 % glucose). All diets were isoenergetic and isonitrogenated, with a standard amino acid content. The diets were assayed in 93 rats with open femoral fracture immobilized by Kirschner pin insertion. The diets were administered for 4 days. On the fifth day, liver and jejunal mucosa protein synthesis were determined. Highest liver protein synthesis rates were obtained with the diet compositions: lipid/carbohydrate ratio: 25% lipids and 75% carbohydrates (expressed as energy ratio). A higher proportion of lipids significantly decreases liver protein synthesis (p <0.05). Jejunal mucosa protein synthesis followed the same pattern, with the same statistical differences. PMID- 9728507 TI - Effect of glycosylated and non-glycosylated prolactin on the proliferation of normal human lymphocytes. AB - We evaluated the effects of 2.5x10(-10) M or 5x10(-10) M concentrations of human pituitary prolactin (pPRL), human recombinant non-glycosylated (NG-PRL) and glycosylated (GL-PRL) prolactin on the proliferation of normal human lymphocytes with or without coactivation by interleukin-2 (IL-2). None of the PRL forms alone affected the lymphocyte proliferation in a serum-free medium, however, the stimulatory activity of IL-2 was significantly potentiated with all 3 PRL variants. Since the 5x10(-10) M concentrations of individual PRLs exerted the same effects, this result suggests that GL-PRL in primary lymphocyte culture is not a less mitogenic form, if sufficient amounts of IL-2 are available. PMID- 9728506 TI - Effects of immunomodulators on postirradiation recovery in the thymus. AB - The effect of immunomodulatory agents on reparation processes in the thymus was studied in mice injured by a single sublethal or lethal dose of ionizing radiation ranging between 6.5-9.5 Gy. Reparation of thymus weight was not influenced by pretreatment with immunomodulators. Furthermore, the morphological picture did not exhibit appreciable differences between non-protected and protected groups, except for greater proliferation of fibroblasts and macrophages in protected animals. PMID- 9728508 TI - Effects of drugs acting on adrenergic and adenosine receptors on the intraocular pressure and the activity of adenylyl cyclase in ciliary processes and their sensitivity to pertussis toxin. AB - The effects of the selective alpha2-adrenergic agonist p-aminoclonidine, the nonselective adrenergic agonist epinephrine, the selective beta2-adrenergic agonist fenoterol and the adenosine A1 agonist R-PIA on intraocular pressure were studied in control and pertussis toxin-pretreated rabbits. Pretreatment of rabbits with pertussis toxin decreased the ocular hypotensive effects of p aminoclonidine and epinephrine, did not influence the same effects of fenoterol or R-PIA and markedly potentiated the initial ocular hypertensive effects of epinephrine and R-PIA. As far as the action on adenylyl cyclase in ciliary processes is concerned, isoproterenol stimulated its activity in control rabbits and epinephrine exerted dual, i.e. stimulatory and inhibitory effects on the activity of this enzyme. The data obtained with epinephrine and p-aminoclonidine confirm the view that their ocular hypotensive effects are associated with their inhibitory action on adenylyl cyclase and contradict the opinion that the hypotensive action of adrenergic drugs depends on adenylyl cyclase activation. PMID- 9728509 TI - Effect of yeast culture and phenolic acids on the physiology of rumen fermentation determined in vitro. AB - The effect of p-hydroxybenzoic acid (HBA), syringic acid (SYA) and yeast culture (YS) on rumen fermentation in vitro has been investigated. Meadow hay was used as a substrate and rumen fluid as an inocula. The yeast culture Levucel contained 5x10(8) yeast cells Saccharomyces cerevisiae per 1 g of dry matter and was used in the amount of 0.5 g/l of the medium. The following combinations of additives were used: hay without additive, hay + YS, hay with 1, 5 or 10 mmol HBA or SYA, and hay + YS with 1, 5 or 10 mmol HBA or SYA. The test tubes were incubated for 96 hours at 39 degrees C. The results showed that 1 mmol HBA had a significant effect on yeast efficacy. This was manifested in the increased degradability of hay dry matter (P <0.05) and enhanced total gas production (P<0.05). SYA in the same amount combined with yeast had a similar effect on gas production (P<0.05), but hay dry matter degradability was not affected. The results showed a slight effect of phenolic acids and yeast culture on hay rumen fermentation in vitro. PMID- 9728510 TI - Projections of auditory cortex onto the inferior colliculus in the rat. AB - The organization of the neocortical projection to the inferior colliculus (IC) was studied in 36 rats using retrograde transport of horseradish peroxidase (HRP) or horseradish peroxidase conjugated with lectin (WGA-HRP). Projection to the external and dorsal cortices originates in the temporal neocortical areas Te 1, Te 2 and Te 3 and in the parietal area Par 2. The corticocollicular projection is predominantly ipsilateral with a weak contralateral contribution. Projection to the rostromedial and rostrolateral part of the external cortex (EC) of the IC arises mainly from the areas Par 2 and Te 1. The participation of the cortical areas Te 2 and Te 3 in this projection is only small. The fibres to the caudobasal part of the external cortex descend from the caudal parts of areas Te 1, Te 2, and Te 3. The corticocollicular projections to the dorsal part of the IC are more numerous than the projections to the EC and originate in all temporal areas, i.e. in area Te 1, Te 2 and Te 3. However, the topographical organization of the corticocollicular projection is more pronounced in the part which projects to the EC. We suggest that the topographical organization of the projections to the EC corresponds with the map of auditory space in the EC. The source of corticocollicular fibres are exclusively neurones of lamina V of all cortical areas sending their fibres to the IC. PMID- 9728511 TI - Application of computer-controlled, real-time TV single-object tracking in behavioural biology and rehabilitation medicine. AB - A PC-based system with TV input for automatic tracking of a single and contrast object in 2D in a homogeneous and stationary environment has been developed and applied to Morris water maze experiments. Further development of the system aimed at broader support of experiments, reduction of requirements on the stationarity and homogeneity of the scene background and on multiple-object tracking is discussed. The computer control of active light markers of the tracked object applicable to multiple-objects tracking in a time-sharing regime is also mentioned in the conclusion. The latter extension of the system can be applied to kinematic studies in biomechanics, sport and rehabilitation medicine. PMID- 9728512 TI - Induction of cell-cycle inhibitor p21 in rat ventricular myocytes during early postnatal transition from hyperplasia to hypertrophy. AB - To examine a possible involvement of p21 protein, an inhibitor of cyclin dependent kinases (CDKs), in the transition from hyperplastic to hypertrophic growth of rat ventricular myocytes during the first postnatal week, we analysed day-by-day changes in the number of p21 positive cells using specific antibodies against this protein. Paraffin-embedded sections of the left ventricular myocardium were examined by means of immunoperoxidase technique and hematoxylin eosin counterstaining. While during the first three postnatal days, the positive reaction for p21 was detected only in a small fraction of myocytes (12-20%), a sudden increase in positivity occurred on day 4 (54%) and continued till day 6 when the fraction of cells expressing p21 reached 87%. Our results show that the induction of CDK inhibitor p21 in rat ventricular myocytes is developmentally regulated. Moreover, the fact that the sudden increase in p21 positivity occurred at the same stage when the myocyte proliferation rapidly ceases, suggests that this protein is likely to be involved in mediating this key event of cardiac development. PMID- 9728513 TI - The effect of triiodothyronine on cell oxidative capacity in regenerating rat liver. AB - The recovery of total DNA content and recovery of total cytochrome c oxidase activity in the rat liver after partial hepatectomy is accelerated by triiodothyronine applied in three doses, two before and one immediately after liver resection. Triiodothyronine-treated animals already have higher cytochrome c oxidase activity before resection. The recovery of the tissue oxidative capacity after partial hepatectomy is more rapid in triiodothyronine-treated animals. These data indicate that hormonal activation of the liver regeneration process is involved. PMID- 9728514 TI - The effect of antigen stimulation on splenic transplants. AB - The present paper deals with the regeneration of splenic tissue after autologous transplantation. Control and transplanted rats (60 days after operation (10(6) cells per injection). The effect of a primary response was studied by a single injection, long-lasting bacteraemia was imitated by 5 injections in weekly intervals. Spleens and transplants were investigated by flow-cytometry and immunohistochemistry. Additionally, the proliferation activity and the specific antibody production against Escherichia coli proteins were tested. Flow cytometric analysis showed altered behaviour of T-helper cells and B-cells in transplants following a primary response, whereas in the multiple injection group a difference between the splenic and transplant response was restricted to macrophages and MHC II+ cells. The results of the morphometric analysis revealed that the cellular composition of unstimulated transplants was very similar to that of the spleen with some subtle alterations. Only the marginal zone showed more striking differences concerning the homing of several cell classes. Under stimulatory conditions, these subtle alterations became more drastic so that CD5+ cells, B-cells and macrophages responded in an abnormal manner in both groups. The analysis of thymidine kinase disclosed decreased activity in the spleen after weekly antigen stimulation. The stimulation index of all transplant groups was significantly lower than that of the spleen. The specific antibody (IgG) production after a single immunization was highest in the transplant group. All groups responded after the multiple challenge. In conclusion, the results demonstrate that splenic transplants differs in several, but subtle aspects from normal splenic tissue. The main reason for most of these alterations may be a slightly misguided recirculation and/or homing of cells. PMID- 9728515 TI - Mesenteric arteriovenous differences in glucose, lactate and insulin concentrations in rats and humans. AB - The present study was aimed to investigate the mesenteric arteriovenous differences in blood glucose and lactate and plasma insulin in humans (n=8) and rats (n=10). Arterial (abdominal aorta) and mesenteric vein blood glucose and lactate (enzymatic methods) and plasma insulin concentrations (radioimmunoassay) were measured in patients during abdominal surgery and in normal rats. Blood glucose levels were significantly (p < 0.05) higher in the abdominal aorta than in the mesenteric vein in both rats (9.2+/-1.0 vs 7.5+/-0.8 mmol/l) and humans (10.4+/-2.9 vs 8.5+/-2.7 mmol/l). Blood lactate levels were higher (p < 0.05) in the mesenteric vein in both rats (3.7+/-1.3 vs 2.8+/-0.9 mmol/l) and humans (0.7+/-0.23 vs 0.1+/-0.05 mmol/l). Plasma insulin concentrations were identical in the aorta or mesenteric vein in both rats (314.4+/-162.0 vs 311.4+/-94.2 pmol/l) and humans (62.4+/-43.2 vs 61.8+/-48.0 pmol/l). In conclusion, both rat and human intestine retained a high proportion of arterially administered glucose and released lactate under the studied conditions. PMID- 9728516 TI - Possible participation of EDRF-NO in the hormonal regulation of bone blood flow in rats. AB - An increase in bone blood flow (BBF) was observed in rats after castration whereas a decrease in BBF occurred after oestradiol or testosterone. The possible participation of prostaglandins in these changes was demonstrated. The present results show that the endothelium-derived relaxing factor, i. e. nitric oxide (EDRF-NO), might play a role in these hormonal actions on BBF. Until now, almost nothing is known about the possible action of NO on bone circulation. Methylene blue (MB) as a substance blocking EDRF-NO was administered to sham-operated or oophorectomized (OOX) female rats. We determined local blood flow (85Sr microsphere uptake), cardiac output, blood pressure, heart rate, density of the tibia and ash weight, as well as 24-h incorporation of 45Ca and 3H-proline into the tibia. The administration of MB (0.5% in the food for 4 weeks) significantly lowered both 85Sr-microsphere uptake and blood flow values in the tibia and distal femur of sham-operated and OOX rats. MB lowered cardiac output and blood pressure to the same extent, indicating no change in the vascular resistance. After the administration of MB (0.1% in the food), 85Sr-microsphere uptake decreased significantly in the tibia of OOX females while no significant change was found in soft tissues. Bone density and ash weight were significantly lower in OOX rats and in sham-operated rats after MB treatment. Finally, the 24-h incorporation of both 45Ca and 3H-proline decreased significantly in OOX females after MB administration (0.04% in the food). It can be concluded that 1) MB lowers BBF, suggesting the participation of EDRF-NO in BBF regulation, 2) MB does not influence or may even suppress cardiac output and blood pressure in high dosage, 3) MB lowers 24-hour incorporation of 45Ca and 3H-proline into the tibia of OOX rats, which is in agreement with the circulatory effect, 4) MB lowers bone density and ash weight of the tibia in non-castrated female rats. The effects of MB observed in our experiments partially differ from those of arginine-derived blocking agents. This requires further elucidation. PMID- 9728517 TI - Relationship between the duration of the breast-feeding period and the lipoprotein profile of children at the age of 13 years. AB - Selected parameters of lipid metabolism were studied in a group of 76 children aged 12-13 years. The children were divided into 4 subgroups according to the duration of neonatal nutrition (no breast feeding, breast feeding for 3, 6 or more than 6 months). We studied the concentration of total serum cholesterol, its distribution into lipoprotein fractions, the concentration of serum triacylglycerols and apolipoproteins A1 (Apo A1) and B (Apo B). Atherogenic indexes were calculated from the data obtained. The highest cholesterol levels (5.20+/-0.15 mmol x l(-1)) were found in children who had been breast-fed for more than 6 months, while the highest concentrations of Apo B (0.80+/-0.07 g x l( 1)) and Apo A1 (1.76+/-0.06 g x l(-1)) and the highest Apo B/Apo A1 ratio (0.45+/ 0.04) were found in children with the shortest period of breast-feeding. No significant sex-related differences in total, VLDL, LDL, HDL cholesterol, triacylglycerols and apolipoproteins were observed. PMID- 9728518 TI - Baroreflex sensitivity during psychological stress. AB - The aim of this study was to analyse the changes of baroreflex sensitivity (BRS) and their relation to changes of heart rate and blood pressure in medical students during moderate psychological stress brought about by oral examination. The changes of BRS during the stress were compared with the changes during light physical exercise. Thirty three students were examined 30 min before and 30 min after the exam. Thirty-nine students of control group were examined at rest and during light exercise. Blood pressure was noninvasively recorded by Penaz method at rate-controlled breathing (0.33 Hz). The BRS [ms/mm Hg] and BRSf [Hz/mm Hg] were calculated by spectral analysis of spontaneous fluctuations of blood pressure and inter-beat intervals (IBI). BRS before examination (7.12 ms/mm Hg) was significantly lower than after the oral exam (8.77 ms/mm Hg, p<0.05). The difference between BRS in the test group after the oral exam and the control group at rest (10.78 ms/mm Hg) was not significant. BRS during light exercise (7.44 ms/mm Hg) corresponded to the value during psychological stress. The values of BRSf did not change during psychological stress (before: 0.0182 Hz/mm Hg; after: 0.0182 Hz/mm Hg) and exercise (rest: 0.0158 Hz/mm Hg; exercise: 0.0144 Hz/mm Hg). Correlation between BRS or BRSf and blood pressure were not found. A significant negative correlation (r=-0.404, p<0.05) between BRSf and the increase of diastolic blood pressure during stress was observed. It is concluded that BRSf remained constant during psychological stress and exercise, and differed essentially from that in hypertensive subjects. PMID- 9728519 TI - Production of hydrogen peroxide by peritoneal macrophages from rats exposed to subacute and chronic hypoxia. AB - Hydrogen peroxide production was measured in non-elicited rat peritoneal macrophages using luminol-dependent chemiluminescence (LDCL). Isolated cells were activated by a chemotactic peptide (FMLP) or by a phorbol ester (PMA) or by the combination of both. A hundred-fold higher LDCL intensity was achieved with PMA relative to FMLP. However, when FMLP was added subsequently to PMA it produced approximately the same response as did PMA. These measurements were carried out with cells isolated from controls and from animals exposed to normobaric hypoxia (10% O2) for 3 hours, 3 days, or 21 days. Hypoxia had a dual effect. Acutely (within 3 hours) it attenuated the production of hydrogen peroxide triggered by PMA, whilst during longer exposure (3 or 21 days) it increased the response induced by FMLP. Hypoxia can thus modulate the capacity of respiratory burst in peritoneal macrophages. PMID- 9728520 TI - The chamber exposure of laboratory rats to metal oxides originating from metal producing industry. AB - Laboratory rats were exposed to the inhalation of dust from an agglomeration unit which is the greatest contributor to dust pollution in the vicinity of a mercury producing plant. The exposure lasted for 6 months (4 hours daily, 5 days per week), the concentration of aerosol in the chamber was 10 mg x m(-3). After finishing the exposure, the animals were examined and compared with the controls which were held under standard laboratory conditions. The number of alveolar macrophages was highly elevated (P< 0.001) in the exposed animals, Mg2+ ATPase activity in the heart muscle was decreased. The alanine aminotransferase activity in the serum was not changed, the aspartate aminotransferase was slightly enhanced. No differences in the frequency of abnormal sperm and in the frequency of polychromatic erythrocytes in bone marrow were detected. PMID- 9728522 TI - Heat-excited and heat-inhibited neurones in the ventroposterior nucleus of thalamus of anaesthetized rats. AB - The activities of 39 single cells, located in the ventroposterior nucleus of the rat thalamus, were recorded from rats deeply anaesthetized with xylazine and ketamine. The activity of each neurone was recorded before and during noxious tail heating. In all, 17 neurones were excited, 11 were inhibited, and 11 were not affected by the noxious stimulation. The possible function of each type of response in the coding of nociceptive information is discussed. PMID- 9728521 TI - Anticonvulsant effect of progabide in rats during ontogenesis. AB - The action of progabide against motor seizures elicited by pentylenetetrazol was studied in 7-, 12-, 18-, 25-day-old and adult rats. Progabide (dissolved in dimethylsulfoxide) was injected in doses from 12.5 to 150 mg/kg i.p. 30 min before pentylenetetrazol. Minimal seizures were not affected by solvent or progabide pretreatment. The action of progabide against major, i.e. generalized tonic-clonic seizures, changed with age: adult rats exhibited a tendency to suppression of whole major seizures, whereas specific suppression of the tonic phase was observed in rat pups during the first three weeks of life. The only effect seen in 25-day-old animals was prolongation of the latency of major seizures after the highest dose of progabide. PMID- 9728523 TI - Phospholipids and calcium alterations in platelets of schizophrenic patients. AB - Alterations in phospholipid metabolism in blood elements have been proposed as the possible biochemical marker of schizophrenia. In the present study, we investigated the composition and membrane distribution of phospholipids in platelets of drug-free schizophrenic patients and controls. We have demonstrated that platelets of drug-free schizophrenics have significantly higher cytosolic Ca2+ levels in comparison with healthy controls. Platelets of drug-free schizophrenic patients have a lower content of phosphatidylinositol (PI). After thrombin activation, PI is the target of phospholipase C instead of phosphatidylinositol 4,5-bisphosphate (PIP2), which is hydrolyzed in platelets of controls. Alterations in the distribution of phospholipids were found in the plasma membrane of platelets of schizophrenic patients. We suggest that alterations in phospholipid metabolism might be evoked by a disturbance of calcium homeostasis in schizophrenic patients. PMID- 9728524 TI - Changes in physical state of ovarian membranes during pseudopregnancy in the rat. AB - The role of the physical state of ovarian membranes was studied in immature rats after PMSG- and hCG-induced pseudopregnancy. Parallel changes in LH/hCG receptors, progesterone secretion and rigidity of membrane lipids were observed during pseudopregnancy. Possible structure-functional properties of the LH/hCG receptor were analyzed by the thermal perturbation technique. Thermal stability of the receptor was higher 5 days after hCG ovulatory injection to rats compared to days 11 or 18 and to control rats. Pseudopregnancy modified the quenching of protein fluorescence. The Stern-Volmer constants for controls and for rats on days 5, 11 and 18 of pseudopregnancy were found to be 2.4 and 4.6, 5.1 and 4.4, respectively, indicating that accessibility of fluorophores for the quencher was increased. PMID- 9728525 TI - Anticonvulsant effect of SL 75 102 in adult and immature rats. AB - The anticonvulsant action of SL 75 102, a metabolite of Progabide, was studied in a model of pentylenetetrazol-induced motor seizures in adult and 12-day-old rats. SL 75 102 suppressed generalized tonic-clonic seizures in adult rats and restricted the tonic phase of these seizures in rat pups. SL 75 102 was less effective than Progabide. In addition, some minor differences in anticonvulsant actions of these two drugs were observed. PMID- 9728526 TI - Valaciclovir for the suppression of recurrent genital herpes simplex virus infection: a large-scale dose range-finding study. International Valaciclovir HSV Study Group. AB - A randomized, double-blind study of valaciclovir for suppression of recurrent genital herpes was conducted among 1479 immunocompetent patients. Patients were randomized to receive valaciclovir (250 mg, 500 mg, or 1 g once daily, or 250 mg twice daily), acyclovir (400 mg twice daily), or placebo, for 1 year. All valaciclovir dosages were significantly more effective than placebo at preventing or delaying recurrences (P < .0001). There was a dose-response relationship (P < .0001) across the once-daily valaciclovir regimens. Twice-daily valaciclovir and acyclovir were similar in effectiveness. Subgroup analysis showed that patients with a history of < 10 recurrences per year were effectively managed with 500 mg of valaciclovir once daily. One gram of valaciclovir once daily, 250 mg of valaciclovir twice daily, or 400 mg of acyclovir twice daily were more effective in patients with > or = 10 recurrences per year. Safety profiles of all treatments were comparable. Thus, valaciclovir is highly effective and well tolerated for suppression of recurrent genital herpes. Once-daily regimens offer a useful option for patients who require suppressive therapy for management of genital herpes. PMID- 9728527 TI - Antibody response and protective capacity of plasmid vaccines expressing three different herpes simplex virus glycoproteins. AB - Plasmid expression vectors were constructed that contained the genes encoding herpes simplex virus 1 (HSV-1) glycoproteins C (gC), D (gD), and E (gE). Mice receiving two intramuscular injections of expression plasmid (50 microg) produced a specific HSV-1 antibody response. Mice receiving the gD plasmid were protected against a lethal intraperitoneal challenge of HSV-1 (5 x 10(4) pfu) but not against more demanding challenge doses. Protection with gC or gE plasmid vaccination could be demonstrated only if the inoculating dose of DNA was increased to 250 microg. In contrast, all mice immunized with vaccinia recombinants expressing either gC or gE survived HSV-1 challenge. Analysis of the HSV-1 antibody isotype produced by plasmid immunization revealed a response dominated by IgG2a. Combination delivery of all three glycoprotein expression plasmids provided better protection against lethal challenge, but mice receiving the combination were still not able to withstand increased challenge doses of virus. PMID- 9728528 TI - Recurrent acyclovir-resistant herpes simplex in an immunocompromised patient: can strain differences compensate for loss of thymidine kinase in pathogenesis? AB - To investigate how acyclovir-resistant (ACVr) herpes simplex virus (HSV) evades drug therapy and causes disease, HSV-1 isolates from a bone marrow transplant (BMT) patient were studied. The patient developed ACVr disease after an initial BMT and, following a second BMT, reactivated ACVr HSV despite high-dose acyclovir prophylaxis. ACVr isolates from each episode contained the same point mutation in the viral thymidine kinase (tk) gene, documenting the emergence, latency, and reactivation of this mutant. The mutants were exceedingly impaired for TK activity in sensitive enzyme, plaque autoradiography, and drug-susceptibility assays. Nevertheless, these mutants and a tk deletion mutant constructed in the same genetic background reactivated from latency in mouse trigeminal ganglia, in contrast to similar mutants from laboratory strains. It is hypothesized that alleles in the clinical isolate compensate for the loss of TK in this animal model. Such genetic variability may be important for ACVr disease in humans. PMID- 9728529 TI - Quantitative polymerase chain reaction to predict occurrence of symptomatic cytomegalovirus infection and assess response to ganciclovir therapy in renal transplant recipients. AB - Cytomegalovirus (CMV) DNA levels were measured by quantitative-competitive polymerase chain reaction (PCR) in weekly leukocyte samples from 50 renal transplant recipients, including 23 with symptomatic and 27 with asymptomatic CMV infection. Peak and week 4 CMV DNA levels were higher in symptomatic subjects (P = .07 and .02, respectively). In a logistic regression model, the logarithm of the week 4 level independently predicted symptomatic infection (odds ratio, 1.78 for a 1 log10 increase; 95% confidence interval, 1.14-2.78; P = .01). All subjects whose week 4 level exceeded 1000 copies/100,000 leukocytes developed symptoms. In subjects with adequate samples for analysis, CMV levels declined exponentially with ganciclovir treatment, with an average half-life of 3.3 days. Levels exceeding 10,000 copies were associated with prolonged time to clearing of CMV DNA. Potential clinical applications of quantitative CMV PCR include predicting occurrence of symptomatic first episodes after transplantation and individualizing duration of antiviral therapy. PMID- 9728530 TI - Changing epidemiology of congenital rubella syndrome in the United States. AB - To describe clinical presentation and epidemiology of US infants with congenital rubella syndrome (CRS) and to identify missed opportunities for maternal vaccination, data from CRS cases reported to the National Congenital Rubella Syndrome Registry (NCRSR) from 1985 through 1996 were analyzed. Missed opportunities for maternal vaccination were defined as missed postpartum, premarital, and occupational opportunities, that is, times when rubella vaccination is recommended but was not given. From 1985 through 1996, 122 CRS cases were reported to the NCRSR. The most frequent CRS-related defect was congenital heart disease. Of the reported infants with CRS, 44% were Hispanic. Of 121 known missed opportunities for rubella vaccination among 94 mothers of infants with indigenous CRS, 98 (81%) were missed postpartum opportunities. CRS continues to occur in the United States. Hispanic infants have an increased risk of CRS. Missed opportunities for postpartum rubella vaccination were identified for 52% of indigenous CRS cases. PMID- 9728532 TI - A mouse model for evaluation of prophylaxis and therapy of Ebola hemorrhagic fever. AB - The Zaire subtype of Ebola virus (EBO-Z) is lethal for newborn mice, but adult mice are resistant to the virus, which prevents their use as an animal model of lethal Ebola infection. We serially passed EBO-Z virus in progressively older suckling mice, eventually obtaining a plaque-purified virus that was lethal for mature, immunocompetent BALB/c and C57BL/6 inbred and ICR (CD-1) outbred mice. Pathologic changes in the liver and spleen of infected mice resembled those in EBO-Z-infected primates. Virus titers in these tissues reached 10(9) pfu/g. The LD50 of mouse-adapted EBO-Z virus inoculated into the peritoneal cavity was approximately 1 virion. Mice were resistant to large doses of the same virus inoculated subcutaneously, intradermally, or intramuscularly. Mice injected peripherally with mouse-adapted or intraperitoneally with non-adapted EBO-Z virus resisted subsequent challenge with mouse-adapted virus. PMID- 9728531 TI - Molecular analysis of rubella virus epidemiology across three continents, North America, Europe, and Asia, 1961-1997. AB - E1 gene nucleotide sequences of 63 rubella virus isolates from North America, Europe, and Asia isolated between 1961 and 1997 were compared phylogenetically. Two genotypes were evident: Genotype I contained 60 viruses from North America, Europe, and Japan, and genotype II contained 3 viruses from China and India. The genotype I isolates prior to 1970 grouped into a single diffuse clade, indicating intercontinental circulation, while most post-1975 viruses segregated into geographic clades from each continent, indicating evolution in response to vaccination programs. The E1 amino acid sequences differed by no more than 3%; thus, no major antigenic variation was apparent. Among 4 viruses from congenital rubella syndrome that occurred following reinfection, only one amino acid substitution occurred in several important epitopes, indicating that antigenic drift is not important in this phenomenon. However, 2 viruses isolated from chronic arthritis exhibited changes in these epitopes. Isolates of the RA 27/3 vaccine strain were readily identifiable by nucleotide sequence. PMID- 9728533 TI - Do dual nucleoside regimens have a role in an era of plasma viral load-driven antiretroviral therapy? AB - This study was undertaken to characterize predictors of response to double nucleoside combinations among 245 human immunodeficiency virus-infected persons initiating antiretroviral therapy. The median time for receiving antiretroviral therapy in this group was 6 months, and the plasma virus load was 58,000 copies/mL. The most commonly prescribed regimens were zidovudine/lamivudine (154 subjects, 63%) and stavudine/lamivudine (46 subjects, 19%). A total of 96 (39%) subjects had their virus load decrease to < 500 copies/mL after the initiation of therapy. Of the 245 study subjects, 102 (41.6%) had > or = 5 months of follow-up and two or more consecutive virus load determinations performed after the start of antiretroviral therapy. Multivariate analysis demonstrated that baseline virus load was the only significant factor associated with obtaining two or more plasma virus loads of < 500 copies/mL. Overall, these data demonstrate that dual nucleoside therapy (using currently licensed agents) cannot reliably achieve a high level of suppression of plasma virus load. PMID- 9728534 TI - Diminished spontaneous apoptosis in lymphocytes from human immunodeficiency virus infected long-term nonprogressors. AB - The relationship between peripheral lymphocyte apoptosis and human immunodeficiency virus disease progression was studied in infected subgroups with distinct profiles of progression. Long-term nonprogressors (LTNP) and seronegative controls had levels of spontaneous apoptosis significantly lower than those for recent seroconverters who had CD4 cell counts similar to those of nonprogressors but with a high likelihood of disease progression. Lymphocytes from nonprogressors and seronegative controls also showed negligible spontaneous caspase-3 activity, a biochemical indicator for apoptosis, whereas early progressors exhibited substantial activity. In contrast, when activated with mitogens, the lymphocytes from both LTNP and progressors displayed indistinguishable levels of heightened apoptosis. Spontaneous apoptosis and plasma viremia levels correlated positively in progressors, but not in LTNP. These findings demonstrate that increased lymphocyte apoptosis is evident prior to CD4 T cell decline and that LTNP are relatively resistant to the factors that induce accentuated levels of spontaneous but not mitogen-induced cell death. PMID- 9728535 TI - Thymic dysfunction and time of infection predict mortality in human immunodeficiency virus-infected infants. CDC Perinatal AIDS Collaborative Transmission Study Group. AB - The effect of human immunodeficiency virus (HIV)-induced thymic dysfunction (TD) on mortality was studied in 265 infected infants in the CDC Perinatal AIDS Collaborative Transmission Study. TD was defined as both CD4 and CD8 T cell counts below the 5th percentile of joint distribution for uninfected infants within 6 months of life. The 40 HIV-infected infants with TD (15%) had a significantly greater mortality than did the 225 children without TD (44% vs. 9% within 2 years). Infants with TD infected in utero had higher mortality than did those infected intrapartum (70% vs. 37% within 2 years), while no significant difference was noted between infants without TD with either mode of transmission. The TD profile was independent of plasma virus load. Virus-induced TD by particular HIV strains and the time of transmission are likely to explain the variation in pathogenesis and patterns of disease progression and suggest the need for early aggressive therapies for HIV-infected infants with TD. PMID- 9728536 TI - Filgrastim restores interleukin-2 production in blood from patients with advanced human immunodeficiency virus infection. AB - Filgrastim induces lymphocytosis, including all T cell subsets, and increased ex vivo interleukin (IL)-2 release as well as lymphocyte proliferation. Since Filgrastim is increasingly used in patients with human immunodeficiency virus (HIV) infection, the effect of Filgrastim on ex vivo cytokine production was determined. Whole blood from 8 healthy volunteers, 5 high-risk volunteers, and 31 HIV-infected outpatients was assayed for cytokine production in response to endotoxin (LPS) or staphylococcal enterotoxin B (SEB) in the presence or absence of 100 ng/mL Filgrastim. LPS-inducible blood cytokine release of HIV-infected patients was not different from that of normal or high-risk volunteers. The suppressive effect of Filgrastim on LPS-inducible blood tumor necrosis factor alpha and interferon-gamma formation in normal volunteers was not found in HIV infected patients. Patients with advanced HIV infection showed reduced IL-2 and IL-4 release in the presence of SEB. In the presence of Filgrastim, IL-2 production was partially restored. PMID- 9728537 TI - Molecular analysis of bovine spongiform encephalopathy and scrapie strain variation. AB - Five mouse scrapie strains, a mouse-passaged scrapie isolate derived from a field case in sheep in Germany, and 2 mouse-passaged bovine spongiform encephalopathy (BSE) isolates were analyzed by immunoblot in regards to banding patterns of proteinase K-digested pathologic prion proteins (PrPres). To obtain reliable results, the photo-imager technique was used for measurement of staining band intensities. Distinct and reproducible profiles were observed for the different strains or isolates. A British and a German BSE isolate were similar, suggesting the same source of infection. The German scrapie isolate resembled scrapie strain ME7, which has frequently been isolated from sheep scrapie in the past. In selected strains or isolates, no influence of the mouse lines used was observed on PrPres profiles, nor were brain region-specific differences apparent. This investigation suggests that PrPres glycotyping can be an invaluable tool for the in vitro differentiation of BSE and scrapie isolates. PMID- 9728538 TI - A conservative amino acid mutation in the chromosome-encoded dihydrofolate reductase confers trimethoprim resistance in Streptococcus pneumoniae. AB - Multidrug-resistant Streptococcus pneumoniae strains have emerged over the past decade at an alarming rate. The molecular mechanism of trimethoprim resistance was investigated in 5 pneumococcal strains isolated in the Washington, DC, area from patients with invasive infections. Cloning and sequencing of the trimethoprim resistance determinant from these pneumococci indicated that an altered chromosome-encoded dihydrofolate reductase (DHFR) was responsible for the observed resistance. Comparison of DHFR sequences from pneumococcal strains with various susceptibilities to trimethoprim, together with site-directed mutagenesis, revealed that substitution of isoleucine-100 with a leucine residue resulted in trimethoprim resistance. Hydrogen bonding between the carbonyl oxygen of isoleucine-100 and the 4-amino group of trimethoprim is proposed to play a critical role in the inhibition of DHFR by trimethoprim. This enzyme-substrate model should facilitate the design of new antibacterial agents with improved activity against S. pneumoniae. PMID- 9728539 TI - Human antibodies elicited by a pneumococcal vaccine express idiotypic determinants indicative of V(H)3 gene segment usage. AB - Human immunodeficiency virus (HIV)-infected persons manifest decreased antibody responses to pneumococcal polysaccharide vaccines. Since human antibody responses to polysaccharides are often restricted, the molecular structure of antibodies elicited by a 23-valent pneumococcal vaccine was analyzed. Anti-idiotypic reagents were used to detect V(H)1, V(H)3, and V(H)4 gene usage by antibodies to pneumococcal capsular polysaccharides in HIV-uninfected and HIV-infected subjects by ELISA. HIV-uninfected persons generated beta-mercaptoethanol-sensitive and resistant antibodies to pneumococcal capsular polysaccharides expressing V(H)3 determinants recognized by the D12, 16.84, and B6 monoclonal antibodies; antibodies expressing V(H)1 determinants were not detected, and V(H)4 determinants were expressed by beta-mercaptoethanol-sensitive antibodies only; and HIV-infected subjects had significantly lower capsular polysaccharide specific and V(H)3-positive antibody responses. These findings confirm decreased antibody responses to pneumococcal vaccination in HIV-infected persons and suggest that their poor responses may result from HIV-associated depletion of restricted B cell subsets. PMID- 9728540 TI - Acquisition versus loss of Helicobacter pylori infection in Japan: results from an 8-year birth cohort study. AB - Studies of the pattern of change in the epidemiology of Helicobacter pylori infection are scarce. A longitudinal cohort study consisted of 644 children and adults, and two independent cross-sectional surveys were conducted in rural Japan between 1986 and 1994. The anti-H. pylori IgG seroconversion rates were 1.1% and 1% per year for children and adults, respectively. The seroreversion rate per year was 1.8% for children and 1.5% for adults. The cohort study was confirmed by the two cross-sectional studies. H. pylori prevalence fell in all age groups in both children (odds ratio [OR] = 0.5, 95% confidence interval [CI] = 0.2-1.0, P = .05) and adults (OR = 0.4, 95% CI = 0.3-0.6, P = .001). The rate of loss of H. pylori infection was greater than the acquisition. Data regarding acquisition and loss of H. pylori infection are critical to understanding the epidemiology of the infection and to developing treatment and vaccination strategies. PMID- 9728541 TI - Pathogenesis of Lyme neuroborreliosis in the rhesus monkey: the early disseminated and chronic phases of disease in the peripheral nervous system. AB - The histopathologic and immunohistochemical features of early and late neuroborreliosis of the peripheral nervous system were investigated in rhesus macaques infected with the JD1 strain of Borrelia burgdorferi. Infection was proven by culture or polymerase chain reaction analysis of skin biopsies and indirectly by Western blot analysis. Three months after infection, neuritis involving multiple nerves was the most consistent neurologic manifestation. Both macrophages and B lymphocytes but not T lymphocytes were present in the cellular infiltrates. Axonal structures surrounding infiltrates had changes consisting of demyelination and axonal phagocytosis. Some of the Schwann cells in lesions stained with anti-nitrotyrosine and anti-tumor necrosis factor-alpha antibodies. B. burgdorferi, or antigens thereof, were visualized immunohistochemically within macrophages. Forty-six months after infection, the most common changes were regenerative, whereas neuritis was infrequent. Aberrant axonal regeneration, irregularly sized myelinated fibers, and fibrosis were frequently observed. Possible mechanisms to explain the appearance and subsidence of Lyme neuritis are discussed. PMID- 9728542 TI - Borreliacidal antibody production against outer surface protein C of Borrelia burgdorferi. AB - Early Lyme borreliosis sera with significant titers of anti-outer surface protein C (OspC) borreliacidal antibodies were identified. Human anti-OspC borreliacidal antibodies could be either IgM or IgG. Significant concentrations of borreliacidal activity were detected after vaccination of mice with OspC. Detection of anti-OspC borreliacidal activity was dependent on surface expression of OspC by Borrelia burgdorferi isolate 50772. The ability of OspC to induce borreliacidal antibodies in vivo and after vaccination offers another possible explanation for the ability of vaccination with OspC to protect against infection with B. burgdorferi. Furthermore, detection of anti-OspC borreliacidal antibodies, especially IgM antibodies, in early Lyme borreliosis sera provides additional evidence that borreliacidal antibody detection may be useful for the serodiagnosis of early Lyme borreliosis. PMID- 9728543 TI - Cytokine and antibody responses in women infected with Neisseria gonorrhoeae: effects of concomitant infections. AB - The levels of interleukin (IL)-1, IL-6, IL-8, IL-10, and transforming growth factor-beta in sera and genital tract secretions from women with gonococcal cervicitis and other genital infections were examined. Cytokines were not elevated in genital secretions from gonococcus-infected compared with uninfected patients. The level of serum IL-6 was higher in gonococcus-infected than in uninfected patients at recruitment. Serum, but not local, IL-1 and IL-6 levels were elevated in patients concomitantly infected with Trichomonas vaginalis or Chlamydia trachomatis in addition to Neisseria gonorrhoeae compared with levels in patients infected with any single organism. Concomitant infection altered neither the total immunoglobulin concentrations nor the levels of antigonococcal antibodies in serum or local secretions. The results suggest that N. gonorrhoeae induces only a limited cytokine and antibody response during uncomplicated cervical infections; however, the presence of other sexually transmitted disease causing organisms can alter the systemic cytokine but not the antigonococcal antibody levels. PMID- 9728544 TI - The role of Gulf Coast oysters harvested in warmer months in Vibrio vulnificus infections in the United States, 1988-1996. Vibrio Working Group. AB - Vibrio vulnificus infections are highly lethal and associated with consumption of raw shellfish and exposure of wounds to seawater. V. vulnificus infections were reported to the Centers for Disease Control and Prevention from 23 states. For primary septicemia infections, oyster trace-backs were performed and water temperature data obtained at harvesting sites. Between 1988 and 1996, 422 infections were reported; 45% were wound infections, 43% primary septicemia, 5% gastroenteritis, and 7% from undetermined exposure. Eighty-six percent of patients were male, and 96% with primary septicemia consumed raw oysters. Sixty one percent with primary septicemia died; underlying liver disease was associated with fatal outcome. All trace-backs with complete information implicated oysters harvested in the Gulf of Mexico; 89% were harvested in water >22 degrees C, the mean annual temperature at the harvesting sites (P < .0001). Control measures should focus on the increased risk from oysters harvested from the Gulf of Mexico during warm months as well as education about host susceptibility factors. PMID- 9728545 TI - Peptide-specific T cell response to Mycobacterium tuberculosis: clinical spectrum, compartmentalization, and effect of chemotherapy. AB - The T cell repertoire of 59 patients with untreated tuberculosis was compared with that of 46 bacille Calmette-Guerin-vaccinated controls by assaying the proliferative responses to six permissively recognized peptides from the 16-, 19 , and 38-kDa molecules of Mycobacterium tuberculosis. A trend from higher to lower reactivity following this order: vaccinated controls > lymph node disease > localized extrapulmonary > pulmonary > pleural was seen for 4 of the peptides (P < .03). The decreased response of blood lymphocytes from patients with pleural tuberculosis was partially accounted for by sequestration of peptide-responsive cells within the pleural fluid. Chemotherapy "reversed" the depressed proliferative responses of patients with pulmonary and pleural tuberculosis depending on the peptide origin, being greatest for peptides of 16 kDa, transient for those of 19 kDa, and least for those of 38 kDa. These data demonstrate antigen specificity in the decreased responsiveness of patients with tuberculosis. PMID- 9728546 TI - Mechanisms of isoniazid resistance in Mycobacterium tuberculosis: enzymatic characterization of enoyl reductase mutants identified in isoniazid-resistant clinical isolates. AB - Mutants in the structural gene of the inhA-encoded NADH-dependent 2-trans enoyl acyl carrier protein reductase were identified from isoniazid-resistant clinical isolates of Mycobacterium tuberculosis. Recombinant InhA proteins with defined single amino acid replacements were expressed in Escherichia coli and purified to homogeneity. Steady-state kinetic parameters for wild type (WT) and I16T, I21V, I47T, and I95P mutants of the enoyl reductase were measured spectrophotometrically. NADH binding to WT and I16T, I21V, I47T, S94A, and I95P mutant reductases were determined by fluorescence spectroscopy and demonstrated that all mutant enzymes had reduced NADH affinity and that NADH binding to all mutants was cooperative as compared with the hyperbolic binding of NADH to the WT enzyme. Since KatG-produced electrophilic derivatives of isoniazid have been suggested to inactivate the enoyl reductase-NADH complex, the kinetics of inactivation for the WT and I21V and I95P mutants was determined. Both mutations resulted in significantly increased values for the apparent first-order rate constant of inactivation. PMID- 9728548 TI - An outbreak of bloodstream infections arising from hemodialysis equipment. AB - An outbreak of 29 cases of bloodstream infection by 16 pathogens occurred during 8 months at two chronic hemodialysis centers. Consequences included 21 hospital admissions and removal of 23 dialysis catheters. An epidemiologic investigation comparing case-patients with uninfected controls showed that risk was significantly (P < .05) associated with having a catheter for vascular access; receiving treatment on a Monday, Wednesday, Friday schedule; and receiving treatment on one heavily contaminated dialysis machine. Culture studies and mock trials showed that bloodstream pathogens were present in a recently installed, commercially marketed attachment for disposal of spent priming saline and could enter blood line tubing directly or indirectly during dialyzer priming and tubing assembly. The outbreak was halted by measures directed at the attachment. Investigation of this problem demonstrated that microbial overgrowth in the attachment caused bloodstream infections and underscores the importance of microbiologic considerations in the design and use of hemodialysis equipment. PMID- 9728547 TI - Genetic diversity among Mycobacterium avium complex strains recovered from children with and without human immunodeficiency virus infection. AB - The genetic diversity and molecular epidemiology of Mycobacterium avium complex (MAC) infections in children with and without human immunodeficiency virus (HIV) infection were evaluated. Isolates recovered from 136 children were subtyped by sequence analysis of a 360-bp region of the gene (hsp65) encoding a 65-kDa heat shock protein. Twenty-one distinct hsp65 alleles were identified. On the basis of hsp65 genotype, 6 isolates were not MAC organisms. Of the remaining 130 samples, 61% were M. avium, 37% were Mycobacterium intracellulare, and 2% were species nonspecific MAC. Eighty-eight percent of the isolates obtained from HIV-infected children were M. avium. In contrast, only 38% of the isolates obtained from children without HIV infection were M. avium (chi2 test, P < .001). M. avium isolates were further subtyped by Southern blot analysis with insertion element IS1245. Taken together, no evidence for a single clonal M. avium strain causing infection was detected. PMID- 9728549 TI - Early signal transduction induced by Candida albicans in macrophages through shedding of a glycolipid. AB - Cell wall beta-1,2-oligomannosides are involved in Candida albicans binding to macrophages and in their stimulation to produce cytokines. The nature of signaling events occurring during initial interaction of macrophage J774 cell line and C. albicans, together with the nature of molecules containing beta-1,2 oligomannosides released by the yeasts, was examined. Cocultivation led to a herbimycin A-sensitive production of tumor necrosis factor-alpha. Immunofluorescence and Western blotting confirmed tyrosine phosphorylation and revealed an accumulation of 90- to 120-kDa phosphoproteins. Antibodies specific for beta-1,2-oligomannosides showed that these epitopes were shed at an early stage from the yeasts to the macrophage membrane, in association with a glycolipid previously described as C. albicans phospholipomannan. Incubation of macrophages with purified phospholipomannan alone led to a signal transduction pathway identical to that observed with living yeasts. All of these results demonstrate that C. albicans phospholipomannan shedding is involved in C. albicans-macrophage interaction through beta-1,2-oligomannosides. PMID- 9728550 TI - Interleukin-15 induces antimicrobial activity after release by Cryptococcus neoformans-stimulated monocytes. AB - A newly described cytokine, interleukin (IL)-15, shares many activities with IL 2; however, little is known about the stimuli for release of IL-15, and its role in antimicrobial host defense has not previously been demonstrated. This study found that Cryptococcus neoformans is a potent stimulus for the release of biologically active IL-15 from monocytes. Both IL-15 and IL-2 made significant contributions to lymphocyte proliferation and lymphocyte-mediated anticryptococcal activity to encapsulated and acapsular C. neoformans. IL-15 restored lymphocyte proliferation and anticryptococcal activity that had been abrogated by blocking IL-2. IL-15 also enhanced the anticryptococcal activity of lymphocytes but did not enhance the activity of monocytes. This suggests that IL 15 and IL-2 cooperate for lymphocyte activation and proliferation in vitro and demonstrates that IL-15 can induce antimicrobial activity. Taken together, these data suggest that microbes, and in particular C. neoformans, are an important stimulus for IL-15 and that IL-15 may have an important role in induction of antimicrobial effector mechanisms. PMID- 9728551 TI - Cysteine proteases of Trichomonas vaginalis degrade secretory leukocyte protease inhibitor. AB - Sexually transmitted diseases, including trichomoniasis, are risk factors for acquisition of human immunodeficiency virus (HIV) infection. Enhancement mechanisms are unknown. Secretory leukocyte protease inhibitor (SLPI) from saliva appears to prevent transmission of HIV through inhibition of virus entry into monocytic cells in vitro. This study was undertaken to determine if secreted cysteine proteases of Trichomonas vaginalis degrade SLPI and render it nonfunctional. It was determined if SLPI levels were decreased in vaginal fluids from pregnant women infected with T. vaginalis. Isolated proteases were incubated with recombinant human SLPI, and the degradation was followed by Western analysis with SLPI antiserum. SLPI levels were measured by ELISA in vaginal fluids from women infected with T. vaginalis and uninfected controls. Cysteine proteases cleaved SLPI and rendered it nonfunctional. Median levels of SLPI from infected patients were 26% of those of controls (P <.005). The degradation of SLPI in association with trichomonal infection may increase the risk of HIV acquisition. PMID- 9728553 TI - The antibody response to 27-, 17-, and 15-kDa Cryptosporidium antigens following experimental infection in humans. AB - Previous studies have suggested that persons infected with Cryptosporidium parvum develop antibody responses to 27-, 17-, and 15-kDa C. parvum antigens. Studies of volunteers infected with Cryptosporidium species provided an opportunity to evaluate the relationship between antibody reactivity to these antigens and infection outcome. As monitored by immunoblot, increases in specific antibody reactivity were more prevalent among volunteers who developed signs and symptoms of cryptosporidiosis (n = 11) than among asymptomatic infected (n = 7; P = .05) or oocyst-negative volunteers (n = 11; P = .02). Volunteers with preexisting IgG antibody to the 27-kDa antigen excreted fewer oocysts than volunteers without this antibody (P = .003). IgG reactivity to the 17-kDa antigens and IgM reactivity to the 27-kDa antigens were higher at day 0 for asymptomatic infected persons than for those who developed symptoms (P = .03 and P = .04, respectively). These results suggest that characteristic antibody responses develop following C. parvum infection and that persons with preexisting antibodies may be less likely to develop illness. PMID- 9728552 TI - Immunologic, microscopic, and molecular evidence of Encephalitozoon intestinalis (Septata intestinalis) infection in mammals other than humans. AB - Encephalitozoon intestinalis (Septata intestinalis) is the second most prevalent microsporidian species infecting humans, but it has not been described in other animal species. This investigation examined 10 domestic animal stool samples (8 mammalian, 2 avian) containing spores detected by anti-Encephalitozoon monoclonal antibody immunofluorescence (FA). The presence of E. intestinalis but not Encephalitozoon hellem or Encephalitozoon cuniculi was confirmed in 6 of 8 mammalian stool samples by species-specific FA and polymerase chain reaction. Clusters of spores inside epithelial cells were observed in feces of five mammals (donkey, dog, pig, cow, and goat) using "quick-hot" Gram-chromotrope stain. None of the 10 samples reacted with anti-E. hellem or anti-E. cuniculi sera, nor were they amplified with species-specific primers for E. hellem and E. cuniculi. To our knowledge, this is the first identification of E. intestinalis in animals other than humans. The data shown herein suggest the possibility that E. intestinalis infection may be zoonotic in origin. PMID- 9728554 TI - Genotypic and phenotypic characterization of Cryptosporidium parvum isolates from people with AIDS. AB - Genotypic analysis of Cryptosporidium parvum has demonstrated the presence of two subgroups within the species, whereas biochemical and antigenic characterization have shown more heterogeneity. The clinical relevance of these observations is unknown. C. parvum isolates from people with AIDS were studied with respect to parasite genotypes and virulence in cell monolayers and laboratory animals. Ten of 13 oocyst samples had a characteristic human-associated (H) genotype; 3 had a genotype typical of calf-excreted oocysts (C). Virulence in cell culture was mildly or markedly lower in the 5 isolates tested (4 H and 1 C) compared with the GCH1 reference isolate. H isolates did not infect newborn ICR mice, whereas 1 of the 2 C isolates tested did. These findings reinforce the concept of C. parvum genetic subgroupings that correlate to some extent with infectivity and suggest that additional heterogeneity is present within the subgroups. PMID- 9728556 TI - Antibodies to Kaposi's sarcoma-associated herpesvirus (human herpesvirus 8) in patients with multiple myeloma. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) serologic assays were used to detect specific antibodies to KSHV lytic and latent antigens in 27 patients with multiple myeloma, 27 control patients with other cancers, and 50 random blood donors. Antibodies to KSHV recombinant minor capsid antigen orf65 were found in 81% of patients with multiple myeloma, 22% of control patients with other cancers, and 6% of the blood donors. Antibodies to KSHV latent nuclear antigens were found in 52% of patients with multiple myeloma, 26% of control patients with other cancers, and 2% of the blood donors. All of the 11 patients with progressive multiple myeloma were KSHV-seropositive. Antibodies to Epstein-Barr virus nuclear antigen 1 were present in 89% of patients with multiple myeloma, 93% of control patients with other cancers, and 88% of the blood donors. These data support the possible association of KSHV infection with multiple myeloma, particularly with progressive disease. PMID- 9728555 TI - Persistence of human herpesvirus 7 in normal tissues detected by expression of a structural antigen. AB - Human herpesvirus 7 (HHV-7) infection in histologically normal human tissues was investigated by immunohistochemical detection of the 85-kDa tegument phosphoprotein (pp85) encoded by the U14 gene. So far, two cell types were recognized as sites of HHV-7 infection in vivo: CD4+ T lymphocytes, believed to be the site of latent infection, and epithelial cells of salivary glands, the site of productive infection and viral shedding. Unexpectedly, cells expressing the HHV-7 structural antigen were detectable in lungs, skin, and mammary glands. Morphologically and phenotypically, they were distinct from lymphocytes. Liver, kidney, and tonsils were positive, although the number of HHV-7-positive cells was low. Large intestine, spleen, and brain were negative. Different from the current notion of the state of HHV-7 in humans, the results show that a variety of tissues harbor cells at a late stage of infection and suggest that HHV-7 causes a persistent rather than a true latent infection. PMID- 9728557 TI - Mucosal and systemic antibody responses to a C4/V3 construct following DNA vaccination of rabbits via the Peyer's patch. AB - A plasmid encoding T1-SP10MN(A), a peptide derived from immunodominant regions of human immunodeficiency virus type 1 gp120, was delivered to rabbit Peyer's patches using a helium-driven gene gun. Six weeks thereafter, 2 of 5 animals were given an intradermal booster immunization. Blood, feces, and vaginal washes were collected weekly and assayed by ELISA. High titer T1-SP10MN(A)-specific fecal and vaginal secretory IgA responses were observed, and the response appeared to be augmented following dermal booster immunizations. Specific serum IgG was also detected within 1 week of immunization and remained elevated through week 20 in the 2 animals receiving dermal boosts (titers > or = 6400). This study establishes the Peyer's patch as a promising target tissue for DNA vaccination and demonstrates the efficacy of gene gun-mediated delivery of foreign DNA to a mucosal tissue for the induction of an immune response. PMID- 9728558 TI - Presence of matrix metalloproteinase-9 activity in the cerebrospinal fluid of human immunodeficiency virus-infected patients. AB - To determine whether matrix metalloproteinase (MMP)-9 is a potential mediator involved in the frequently detected blood-brain barrier leakage in human immunodeficiency virus (HIV)-infected patients, zymography was used to detect MMP 9 activity in cerebrospinal fluid (CSF) samples of 80 HIV-infected patients and of 10 control patients. CSF MMP-9 activity was detected in 40% of HIV-infected patients (but not in controls) and was significantly more frequent in HIV infected patients than in those without neurologic deficits (50% vs. 13.6%). The frequency of CSF MMP-9 activity did not significantly differ between neurologically symptomatic HIV-infected patients with or without opportunistic central nervous system disease (51.6% vs. 48.1%). Additionally, the presence of CSF MMP-9 activity in HIV-infected patients was associated with an increased CSF white blood cell count and an elevated CSF-to-serum albumin ratio, suggesting that it may play a role in blood-brain/CSF barrier leakage in HIV-infected patients. PMID- 9728559 TI - Administration of imiquimod, an interferon inducer, in asymptomatic human immunodeficiency virus-infected persons to determine safety and biologic response modification. AB - A phase I study to determine safety, maximum tolerated dose, and biologic response during multiple once-a-week administration of oral imiquimod, an immune response modifier, was conducted in 12 adults with early human immunodeficiency virus (HIV) infection. All completed the dose-escalation phase of weekly dosing at 100-mg increments and received at least one maintenance dose, 100 mg below the patient's toxic dose, for 12 weeks. Dose-limiting toxicity occurred in 3 patients at 200-mg, 5 at 300-mg, and 3 at 400-mg dose levels. One tolerated the 500-mg dose without dose-limiting toxicity. Dose-limiting toxicities included fatigue, fever, malaise, increased transaminases, hypotension, vomiting, and depression. Seven of 12 completed 12 weeks of maintenance. At > or = 200 mg of imiquimod, all patients had biologic responses, measured by elevations in serum interferon, beta2-microglobulin, and neopterin levels. Imiquimod induced pronounced levels of circulating interferon in asymptomatic HIV-infected persons, with variable effect on virus load. PMID- 9728560 TI - Hepatitis G virus infection in human immunodeficiency virus type 1-infected mothers and their children. AB - Hepatitis G virus (HGV) RNA and anti-E2 glycoprotein antibody (E2Ab) seroprevalence was studied in 58 human immunodeficiency virus type 1 (HIV-1) infected mothers (34 injecting drug users [IDUs] and 24 with risky sexual behavior [RSB]) and their children (median age, 5 days; range, 1-27). Twelve women (20.6%) were RNA- and 20 (34.4%) E2Ab-positive. Seroprevalence was similar in the IDU and RSB groups and high in RSB partners of IDU men. Five (41.6%) children of RNA-positive mothers were HGV-infected, at a median age of 5 days (range, 1-27), independent of maternal CD4 T lymphocyte numbers, mode of delivery, and HIV-1 transmission; no other child at risk became RNA-positive subsequently. No HGV-infected child (follow-up, 16 months; range, 12-52) showed increased liver enzyme levels; 3 children cleared RNA and E2Ab-seroconverted after 10-48 months. Thus, in HIV-1-infected women, HGV infection is common and also sexually transmitted, and clearance may be impaired. Mother-to-child transmission is frequent and occurs antenatally; children remain long infected without evident disease. PMID- 9728561 TI - Emergence of an S gene mutant during thymosin alpha1 therapy in a patient with chronic hepatitis B. AB - The presence of a hepatitis B virus S gene mutant was investigated in a patient being treated with thymosin alpha1. He was seropositive for hepatitis B e antigen throughout therapy but was intermittently seronegative for hepatitis B surface antigen (HBsAg) by an RIA. Sequence analysis revealed an S gene mutant in HBsAg seronegative serum with two consecutive amino acid substitutions: threonine115-to isoleucine and threonine116-to-asparagine, whereas no amino acid substitution or deletion was found in the pre-S region. A site-directed mutagenesis experiment confirmed that these mutations were responsible for the failure to detect HBsAg. In summary, an S gene mutant was identified in an HBsAg-seronegative patient. The mutations were located outside the putative "a" determinant. The emergence of an S gene mutant during thymosin alpha1 treatment suggests that enhanced host immunity against HBsAg may play a role in its antiviral activity. PMID- 9728562 TI - Induction of immunologic refractoriness in adults by meningococcal C polysaccharide vaccination. AB - Thirty-four adults were vaccinated with 1/50 of the usual dose of meningococcal polysaccharide vaccine (1 microg of A, C, Y, and W135 polysaccharides, given intramuscularly). This dose was selected as a probe to assess B cell memory. The probe elicited meningococcal C bactericidal antibody responses in all 18 adults who had been vaccinated 4 years earlier with an investigational meningococcal A and C oligosaccharide-protein conjugate vaccine and in the majority of the 11 subjects vaccinated for the first time. In contrast, the responses of the 5 adults given a full dose of licensed polysaccharide vaccine 4 years earlier were <1/10 of those of the other 2 groups. Thus, adults previously given a full dose of meningococcal polysaccharide vaccine show evidence of immunologic refractoriness to group C polysaccharide, whereas refractoriness is not observed after conjugate vaccination. These findings have implications for the use of meningococcal polysaccharide vaccine when the risk of disease is low. PMID- 9728563 TI - Interferon-gamma-induced activation of indoleamine 2,3-dioxygenase in cord blood monocyte-derived macrophages inhibits the growth of group B streptococci. AB - Neonatal sepsis is most often caused by group B streptococci (GBS) and is a major cause of death in the neonatal period. The response of the immune system in the newborn child has received much attention and is thought to be deficient in a number of ways. The effector response of neonatal monocyte-derived macrophages (MDM) was investigated. Interferon-gamma induced the activation of indoleamine 2,3-dioxygenase in MDM and inhibited the growth of GBS. Both effects were enhanced by the addition of tumor necrosis factor-alpha to the culture conditions. The coincident supplementation of L-tryptophan with the bacteria abrogated the bacterial growth inhibition, thus confirming the causative role of L-tryptophan depletion. Control of the extracellular as well as intracellular L tryptophan levels may thus be one of the effector mechanisms with which the immune system defends the host against GBS dissemination and disease. PMID- 9728564 TI - Anticapsular polysaccharide antibodies and nasopharyngeal colonization with Streptococcus pneumoniae in infant rats. AB - To evaluate the effect of passive immunization with anticapsular antibodies on nasopharyngeal carriage, two models of Streptococcus pneumoniae colonization were developed in infant rats. In a direct inoculation model, 3- to 4-day-old infant rats were intranasally inoculated with 2 x 10(5) cfu of S. pneumoniae type 3 or 6 x 10(3) cfu of S. pneumoniae type 23F. In an intralitter transmission model, 2 infant rats were intranasally inoculated with 10(3) cfu of pneumococcus type 3 or type 19F and placed in a cage containing 10 infant rats. Pretreatment with bacterial polysaccharide immune globulin led to a significant reduction in colonization of contact animals with S. pneumoniae type 3 or 19F in the intralitter transmission model (P < .05). No effect of immune globulin could be demonstrated in the direct inoculation model. These results indicate that systemic anticapsular antibodies conferred significant protection against nasopharyngeal acquisition by intralitter spread of S. pneumoniae type 3 and 19F. PMID- 9728565 TI - Lymph of patients with a systemic inflammatory response syndrome inhibits lipopolysaccharide-induced cytokine production. AB - In patients with systemic inflammatory response syndrome (SIRS), tolerance of peripheral blood mononuclear cells to a second challenge with lipopolysaccharide (LPS) has been described. Thoracic duct lymph transports LPS and represents the extravascular, interstitial fluid compartment of the body. The aim of this study was to determine the capacity of lymph to influence LPS-induced cytokine production in vitro. Thoracic duct lymph was obtained from patients with SIRS and without SIRS (controls). The effect of lymph and simultaneously collected plasma on LPS-induced cytokine production by normal peripheral blood mononuclear cells was assessed. Both lymph and plasma of patients with SIRS reduced LPS-induced tumor necrosis factor-alpha and interleukin-6 production (P < .01); lymph of controls also inhibited cytokine production (P < .01), although to a lesser extent. This study suggests that LPS tolerance may occur both in the intra- and extravascular compartments. PMID- 9728566 TI - Circulating leptin levels during acute experimental endotoxemia and antiinflammatory therapy in humans. AB - Leptin, a newly discovered adipose tissue-derived weight-reducing hormone, is increased in acute inflammation and may be involved in the anorexia and wasting syndrome associated with infection. To determine whether this hormone responds to an acute inflammatory stimulus, plasma leptin concentrations were measured in 12 healthy subjects after intravenous administration of endotoxin. These subjects were randomized to receive concurrently ibuprofen or placebo normal saline (6 in each group). Endotoxin administration resulted in fever, leukocytosis, and an increase in plasma levels of the stress hormones adrenocorticotropic hormone (3.2 +/- 0.3 to 132.6 +/- 75.5 pmol/L, P = .001) and cortisol (431.6 +/- 44 to 796.9 +/- 99 mmol/L, P = .001). Plasma leptin levels, however, did not change significantly from baseline values after administration of endotoxin (0 h: 6.9 +/ 3.1 ng/mL; 6 h: 6.0 +/- 2.2; 24 h: 6.5 +/- 2.8). While ibuprofen suppressed fever and symptoms associated with endotoxemia, it had no effect on the plasma levels of leptin. In conclusion, acute experimental human endotoxinemia is not associated with acute changes in circulating leptin levels. PMID- 9728567 TI - Inhibition of neutrophil apoptosis and modulation of the inflammatory response by granulocyte colony-stimulating factor in healthy and ethanol-treated human volunteers. AB - Granulocyte colony-stimulating factor (G-CSF) has immunomodulating properties that could be beneficial for adjunctive treatment of severe infections. Cytokine release from stimulated whole blood and expression of neutrophil surface and apoptosis markers in response to G-CSF were studied in human volunteers under physiologic conditions and after ethanol pretreatment. Levels of interleukin (IL) 1 receptor antagonist and soluble tumor necrosis factor (TNF) receptor-1 were significantly increased after G-CSF, whereas TNF-alpha and IL-10 concentrations were reduced, and IL-1beta and IL-8 remained unchanged. There was a significant inhibition of neutrophil apoptosis and increased expression of complement regulatory protein CD55 without changes in CD11b, CD14, and CD59 expression. These effects were well preserved after ethanol pretreatment, which per se led to an increase in apoptosis and decreased CD55 expression. Thus, G-CSF treatment was associated with a reduction of the proinflammatory cytokine response and enhanced neutrophil survival in vivo, suggesting a therapeutic potential of G-CSF for severe infections in the nonneutropenic host. PMID- 9728568 TI - Clarithromycin significantly improves interleukin-12-mediated anti-Mycobacterium avium activity and abolishes toxicity in mice. AB - Treatment of experimental murine Mycobacterium avium (MAC) infection with interleukin-12 (IL-12) significantly decreased MAC organisms in tissue but resulted in toxicity. Because IL-12-related toxicity was seen only in infected mice, IL-12 was combined with clarithromycin in an attempt to decrease bacterial burden. Clarithromycin (200 mg/kg/day) was administered alone to M. avium infected mice for 1 week, and from week 2, IL-12 (20 microg/kg twice per week) was added to the regimen for 4 weeks. Treatment with IL-12 resulted in 60% mortality, compared with 40% mortality in untreated control mice and 20% when IL 12 was given with clarithromycin (P < .05). Clarithromycin plus IL-12 resulted in increased activity compared with either clarithromycin or IL-12 alone in reducing the number of bacteria in spleen and blood. Although potentially toxic, IL-12 is an effective immunotherapy for MAC infection, and combination with clarithromycin reduces IL-12 toxicity. PMID- 9728569 TI - Combination drug therapy for cryptosporidiosis in AIDS. AB - Aside from effective antiretroviral therapy, there is no consistently effective antiparasitic therapy for cryptosporidiosis in AIDS. The purpose of this study was to assess safety, efficacy, and durability of combination therapy with paromomycin and azithromycin for chronic cryptosporidiosis. Patients with AIDS, chronic cryptosporidiosis, and < 100 CD4 cells/microL were treated with open label paromomycin (1.0 g twice a day) plus azithromycin (600 mg once a day) for 4 weeks, followed by paromomycin alone for 8 weeks. In 11 patients, median stool frequency decreased from 6.5/day (baseline) to 4.9/day (week 4) and 3.0/day (week 12). Median reductions in 24-h oocyst excretion were 84%, 95%, and >99% at 2, 4, and 12 weeks, respectively. None of the responses were attributable to antiretrovirals. Of 5 survivors at 12-30 months of follow-up, 3 remain asymptomatic off medications, and 2 have chronic, mild diarrhea. Treatment of cryptosporidiosis with azithromycin and paromomycin was associated with significant reduction in oocyst excretion and some clinical improvement. PMID- 9728570 TI - Risk factors for intestinal microsporidiosis in patients with human immunodeficiency virus infection: a case-control study. AB - A prospective unmatched case-control study was conducted to determine risk factors for intestinal microsporidiosis in persons infected with human immunodeficiency virus (HIV) who had < or = 200 CD4 cells/mm3. In multivariate analysis, case-patients (n = 30) were more likely than were control-subjects (n = 56) to have < or = 100 CD4 cells/mm3 (odds ratio [OR], 6.5; 95% confidence interval [CI], 1-42), to report male homosexual preference (OR, 7.6; 95% CI, 1 59.5), and to report swimming in a pool in the previous 12 months (OR, 9.2; 95% CI, 2.1-38.9). In summary, intestinal microsporidiosis in persons with HIV infection and < or = 200/mm3 CD4 cells is associated with male homosexuality and swimming in pools, suggesting fecal-oral transmission, including sexual and waterborne routes. PMID- 9728571 TI - Therapeutic effect of interferon-gamma gene transfer in experimental visceral leishmaniasis. AB - Interferon (IFN)-gamma, in both natural endogenous form and administered as exogenous protein, induces control over visceral Leishmania donovani in experimentally infected BALB/c mice. To further characterize the therapeutic role of IFN-gamma in host defense against intracellular L. donovani, the efficacy of IFN-gamma delivered by gene transfer was tested. One week after infection, normal and IFN-gamma gene-disrupted (GKO) BALB/c mice were injected with an IFN-gamma gene-bearing mammalian expression plasmid (pIFN). Plasmid-specific IFN-gamma transcripts were detected in liver and spleen. Whereas liver parasite burdens more than doubled in untreated and mock-treated normal and GKO mice during the subsequent 2 weeks, animals injected with pIFN had controlled visceral infection and reduced parasite burden. These results indicate that, in infected tissues, IFN-gamma delivered by gene transfer enhances control over disseminated intracellular infection. PMID- 9728572 TI - Detection of Leishmania DNA by polymerase chain reaction in scars of treated human patients. AB - Leishmaniasis is an endemic disease in developing countries. The efficacy of therapy is usually evaluated through clinical parameters. To define the parasitologic cure, 20 patients were biopsied before and 1 month to 8 years after treatment. Paraffin-embedded tissue was used for DNA isolation. All patients had a positive polymerase chain reaction (PCR) result before therapy, except 1, for whom no histopathologic material was available. The causative agent was identified as belonging to the Leishmania (Viannia) subgenus by hybridization. Despite clinical healing and absence of reactivation or development of mucosal lesions, PCR was positive in scars of 16 patients (80%). The results suggest that parasites persist in the skin for many years despite treatment. Depending on specific pathogenetic features of the parasite and the immune status of the host, this phenomenon might result in mucosal lesions. Alternatively, it could have a role in the maintenance of immunologic memory in patients living in areas in which leishmaniasis is endemic. PMID- 9728573 TI - Treatment for AIDS-associated cryptosporidiosis. PMID- 9728574 TI - Perspective: Attributable mortality--the promise of better antimicrobial therapy. PMID- 9728575 TI - Strains of Escherichia coli belonging to serogroups O157 and O55 express lipopolysaccharides that are structurally distinct and do not share common epitopes. PMID- 9728576 TI - Use of statistics and scientific inference: odds ratios, likelihood ratio, and receiving operating characteristic curves. PMID- 9728577 TI - Lung Cancer Experts' Symposium. Proceedings. Stresa, Italy, October 1997. PMID- 9728578 TI - An update on North American randomized studies in non-small cell lung cancer. AB - Lung cancer is the leading cause of cancer-related death in North America and Europe. Approximately 75% of lung cancer is non-small cell lung cancer, and approximately 70% of these patients present with unresectable disease. In the past, these patients were treated with either palliative radiotherapy or other best supportive care measures. The survival of patients with stage IV disease was extremely poor. Median survival was between 16 and 17 weeks, and only 10% to 15% of patients were alive at 1 year. Cisplatin-based chemotherapy was the first therapy to show that survival could be improved. Randomized trials that compared best supportive care, including palliative radiotherapy, with cisplatin-based combination therapy showed modest improvement in the survival of these patients. On average, the median survival of patients improved by 10 weeks (from 16 to 26 weeks) and the 1-year survival rate improved by 10% (from 15% to 25%). These cisplatin-based therapies also relieved symptoms and improved quality of life at acceptable costs, which were sometimes less than those associated with best supportive care. More recently, a number of new chemotherapeutic agents (gemcitabine, paclitaxel, docetaxel, vinorelbine, and irinotecan) with high, single-agent activities and novel mechanisms of action have shown superior activity and improved quality of life when used in combination with conventional agents and with each other. PMID- 9728579 TI - An update on European randomized studies in non-small cell lung cancer. AB - Less than 25% of non-small cell lung cancer (NSCLC) patients present with surgically resectable stage I and II disease, and the majority of patients will eventually die of disseminated disease. Cisplatin-based chemotherapy has been shown to prolong survival and improve quality of life in patients with stage III and IV disease. In recent years, new chemotherapeutic agents (eg, gemcitabine, paclitaxel, docetaxel, vinorelbine, and irinotecan) have shown definite activity in advanced NSCLC. The activity of these agents has led to phase II studies combining them with cisplatin, producing response rates that are generally in excess of 35%. Randomized trials suggest that survival is improved when combinations of gemcitabine/cisplatin, vinorelbine/cisplatin, and paclitaxel/cisplatin are compared with cisplatin alone or cisplatin/etoposide. The following report reviews the ongoing trials in NSCLC. PMID- 9728580 TI - Single-agent gemcitabine versus cisplatin/etoposide in patients with inoperable, locally advanced, or metastatic non-small cell lung cancer. AB - Non-small cell lung cancer (NSCLC) remains a malignancy that is refractory to most currently available chemotherapeutic agents. The most frequently used systemic therapy is cisplatin-based combination therapy, which is not always suitable for patients with stage IV disease because of toxicity. Thus, any single agent with a good patient tolerability profile would be a useful alternative for those patients for whom the main purpose of treatment is palliation. The novel nucleoside analog gemcitabine has proven single-agent activity in patients with NSCLC, with consistent response rates of 20% and above. This report reviews two randomized phase II studies of single-agent gemcitabine versus cisplatin/etoposide in patients with advanced NSCLC. PMID- 9728582 TI - The activity of gemcitabine plus cisplatin in randomized trials in untreated patients with advanced non-small cell lung cancer. AB - Three separate phase III randomized studies were conducted to compare the efficacy of a gemcitabine plus cisplatin combination (GC) with other chemotherapy regimens in the treatment of European and North American patients with inoperable (stage IIIB or IV) non-small cell lung cancer (NSCLC). The Spanish Lung Cancer Group (SLCG) compared the GC regimen with an etoposide plus cisplatin combination (EP), the Hoosier Oncology Group (HOG) compared it with single-agent cisplatin, and the Italian Lung Cancer Project (ILCP) compared it with a mitomycin plus ifosfamide plus cisplatin combination (MIC). The three studies each had a different primary objective (response rate, survival, or quality of life, respectively). From July 1995 to February 1997, 751 patients were enrolled into the three studies. In the SLCG study, 69 were entered in the GC arm and 64 in the EP arm; in the HOG study, 155 were entered in the GC arm and 154 in the cisplatin alone arm; and in the ILCP study, 154 were entered in the GC arm and 152 in the MIC arm. In the SLCG study, gemcitabine 1,250 mg/m2 was given by 30-minute infusion on days 1 and 8 of each 21-day cycle. In the HOG and ILCP studies, gemcitabine 1,000 mg/m2 was given on days 1, 8, and 15 of each 28-day cycle. Cisplatin 100 mg/m2 was given on day 1 in the SLCG and HOG studies and on day 2 in the ILCP study. In the EP arm, etoposide 100 mg/m2 was given on days 1, 2, and 3 of each 21-day cycle. In the MIC arm, mitomycin 6 mg/m2 and ifosfamide 3 g/m2 were given on day 1 of each 28-day cycle. In the SLCG study, 28 patients (40.6%) in the GC arm and 14 patients (21.2%) in the EP arm achieved an objective response (P = .01). In the HOG study, 48 patients (31%) in the GC arm and 14 patients (9.1%) in the cisplatin-alone arm attained on objective response (P < .001). In the ILCP study, 61 patients (40%) in the GC arm and 42 patients (28%) in the MIC arm achieved an objective response (P = .03). Progression-free time was significantly longer for the GC arm (6.8 months) than for the cisplatin-alone arm (4.2 months) (P < .001). The median survival time was remarkably identical for the GC arms in the three studies (8.7 months), whereas it was 7.2 months for the EP arm, 7.6 months for the cisplatin-alone arm, and 9.5 months for the MIC arm. The main toxicities were myelosuppression and vomiting. The assessment of quality of life in the ILCP study is ongoing. Both 21- and 28-day cycles of GC were effective in the treatment of NSCLC, and this combination can be considered as a current "standard" therapy for advanced NSCLC patients. PMID- 9728581 TI - Gemcitabine as second-line treatment for relapsing or refractory advanced non small cell lung cancer: a phase II trial. AB - We investigated the activity and toxicity of gemcitabine as a single agent in patients with advanced non-small cell lung cancer (NSCLC) after recurrence or failure of previous treatment with a platinum-containing regimen. From November 1995 to October 1997, 83 patients (73 men and 10 women) with stage IIIB or IV NSCLC received gemcitabine 1,000 mg/m2 on a weekly x 3 every 4 weeks schedule. Responses were assessed every two treatment courses. The median age of the patients was 63 years. Eastern Cooperative Oncology Group performance status was 0-1 in 62 patients; 2 in 21 patients. The predominant histology was squamous (39 patients); 49 patients had stage IV disease and 34 patients had stage III disease (33 stage IIIB and I stage IIIA). Sixteen patients (19%) achieved a partial response to treatment; the median duration of response was 29 weeks (range, 6 to 50 weeks). Treatment was well-tolerated: leukopenia and thrombocytopenia World Health Organization grade 2-3 occurred in 23% and 20% of patients, respectively. Mild asthenia was observed in 16% of patients and peripheral edema was observed in 5% of patients. Nausea and vomiting were present in 16% of patients. In this study, gemcitabine showed significant activity in a patient population usually associated with poor prognosis. This finding suggests a possible role for gemcitabine as second-line treatment in patients who have recurring disease or who have failed a platinum-containing regimen, and in the absence of significant toxicity. PMID- 9728583 TI - Combined cisplatin and gemcitabine for non-small cell lung cancer: influence of scheduling on toxicity and drug delivery. AB - The scheduling of cytotoxic chemotherapy has important bearing on the toxicity and ability to deliver chemotherapy at or close to full dose. We report new data on the influence of different schedules of cisplatin and gemcitabine on toxicity and drug delivery in phase II studies of non-small cell lung cancer. Patients in six phase II studies had standard entry criteria for advanced non-small cell lung cancer. Whereas gemcitabine was given on days 1, 8, and 15 of a 28-day cycle in all these studies, the scheduling of cisplatin varied and was given either on day 1, day 2, day 15 (in two studies), or days 1, 8, and 15 (in two studies). The protocol dose per infusion for gemcitabine was 1,000 mg/m2 (five studies) and 1,500 mg/m2 (one study); for cisplatin, it was 100 mg/m2 when given once per cycle and 30 mg/m2 when given on days 1, 8, and 15. Similar dose reduction schedules were implemented in the event of grade 3 or higher drug toxicity for all studies except for the day 1 cisplatin study, in which the dose was omitted for grade 2 thrombocytopenia. Nonhematologic toxicity was very low. Hematologic toxicity was moderate, and in patients who developed grade 3 or 4 toxicity, there was no hemorrhage from thrombocytopenia and neutropenic sepsis was rare. The incidence of grade 3 or 4 thrombocytopenia with the day 1, day 2, day 15 (two studies combined), and days 1, 8, and 15 (two studies combined) cisplatin regimens was 50%, 52%, 26%, and 38%. The incidence of grade 3 or 4 neutropenia with these four regimens was 51%, 37%, 56%, and 49%, respectively. Although the hematologic toxicity might appear relatively similar, it represents the toxicity at the administered rather than the intended (protocol) dose, because drug delivery was reduced or omitted in the event of grade 3 or 4 toxicity. Differences between the schedules are revealed by analysis of the actual dosages delivered. The median dosage of gemcitabine per scheduled infusion was statistically higher with the day 15 cisplatin regimens (combined) compared with any of the other regimens treating at 1,000 mg/m2 (P < .003, z-score). The dose with the day 1, day 2, day 15, and days 1, 8, and 15 cisplatin regimens was 664, 829, 889, and 774 mg/m2, respectively. Both the percentages of cycles in which gemcitabine infusions were given at full dose and in which there were no omissions of gemcitabine infusions (including infusions with dose reductions) were statistically higher in the day 15 cisplatin regimen than with any of the other regimens (P < .0001, chi-square test). The percentage of cycles containing full-dose gemcitabine with the day 1, day 2, day 15, and days 1, 8, and 15 cisplatin regimens was 24%, 44%, 75%, and 46%, respectively. The percentage of cycles in which there were no omissions of gemcitabine infusions for the four regimens above was 32%, 55%, 83%, and 72%, respectively. Apart from the once weekly regimen (days 1, 8, and 15) in which the protocol gemcitabine dose was 1,250 mg/m2, the day 15 cisplatin schedule allowed for the highest median concentration of gemcitabine. More importantly, the day 15 cisplatin schedule provided the longest duration of gemcitabine exposure, which is particularly important for its activity as gemcitabine is a phase-specific agent. The day 15 cisplatin schedule is associated with the best dose intensity and the longest median duration of exposure to gemcitabine, and best meets the goal of administering both agents at full doses in combination. PMID- 9728584 TI - Gemcitabine and paclitaxel combinations in non-small cell lung cancer. AB - Gemcitabine and paclitaxel are new cytotoxic agents that have been used both as single agents and in combination, particularly with cisplatin, in the therapy of non-small cell lung cancer (NSCLC) with promising results. The lack of overlapping toxicities and different mechanisms of action of gemcitabine and paclitaxel make the combination of these drugs appealing in the treatment of NSCLC. A number of phase I and II trials are evaluating the use of this combination of cytotoxic agents as first-line therapy and as second-line therapy in patients with advanced NSCLC. In ongoing phase II trials using a 21-day schedule, the combination of gemcitabine and paclitaxel therapy appears to be well-tolerated, with response rates ranging from 29% to 58% and hematologic toxicities that are mild to moderate in severity. Other reported toxicities included alopecia, fatigue, and flu-like symptoms, which were also mild to moderate. However, grade 2/3 neurotoxicity was also reported. In conclusion, preliminary results from these early phase II trials of gemcitabine/paclitaxel combination therapy in advanced NSCLC are encouraging. PMID- 9728585 TI - Gemcitabine and carboplatin in combination: phase I and phase II studies. AB - Gemcitabine has demonstrated single-agent activity in advanced, incurable non small cell lung cancer, yielding response rates of > or =20%, and, in combination with cisplatin, response rates of 30% to 60%. Carboplatin causes much less nonhematologic toxicity than cisplatin, and initial therapy with carboplatin yields equivalent survival in advanced non-small cell lung cancer compared with cisplatin combinations and other cisplatin analogs, despite a lower response rate. Carmichael and colleagues have identified a maximum tolerated dose of carboplatin of area under the curve 5.2 mg/mL/min administered monthly in combination with gemcitabine 1,000 mg/m2 on days 1, 8, and 15. This combination produced a response rate of 31% and a median survival of 45 weeks in 13 evaluable patients. A subsequent phase I evaluation reversing the carboplatin and gemcitabine sequence (gemcitabine --> carboplatin day 1) demonstrated no difference in toxicities, pharmacodynamics, or maximum tolerated dose. At Fox Chase Cancer Center and elsewhere, similar phase I trials will determine the maximum tolerated doses of each agent in combination using a compressed schedule with carboplatin administered every 3 weeks and gemcitabine given on days 1 and 8 only. PMID- 9728586 TI - Triplet chemotherapy combinations with new agents: is there a rationale? AB - Lung cancer remains the major cause of cancer-related death in North America and Europe. Lung cancer causes 28% of all cancer deaths, more than breast, prostate, colorectal, and ovarian cancers combined. Eighty-five percent to 90% of cases of lung cancer are known to be a direct consequence of smoking. Despite this, the incidence of the disease continues to increase dramatically in women, although in the United States, the incidence and mortality rates of lung cancer in men have declined slightly in the last few years. Non-small cell lung cancer (NSCLC) represents 75% to 80% of all primary lung cancers, and approximately 70% of these patients present with unresectable disease. These patients are candidates for palliative radiotherapy and/or chemotherapy, but most of these patients will develop locally advanced or metastatic disease. Currently, the 5-year survival rate across all stages of the disease is 14% in the United States. Until recently, chemotherapy was considered to be more successful in the treatment of small cell lung cancer than NSCLC, but this is no longer true. In a recent meta analysis of randomized trials in NSCLC, cisplatin-based chemotherapy was shown to prolong survival for patients across all stages of the disease. A role for adjuvant cisplatin-based therapy has been shown in early stage disease, and cisplatin-based therapy was shown to improve survival when combined with radiotherapy in locally advanced disease and as a single modality in metastatic disease. Other randomized trials have shown that cisplatin-based therapy improved quality of life in both stage III and IV NSCLC patients and reduced the cost of medical care compared with best supportive care. Cisplatin-based therapy should therefore be considered the standard treatment for all NSCLC patients with locally advanced or metastatic disease. However, over the last 5 years, five new agents have emerged that have increased single-agent response rates, increased survival, and, for the most part, reduced toxicity. The use of these in two-drug combinations with conventional agents will be compared with new two- and three drug combinations. PMID- 9728587 TI - Adjuvant chemotherapy in non-small cell lung cancer. AB - Surgery is the main curative treatment of patients with non-small cell lung cancer (NSCLC), but half of all patients will experience local or distant failure after complete resection. An individual data-based meta-analysis has suggested a 13% reduction in the risk of death and an absolute benefit of 5% at 5 years with adjuvant cisplatin-containing chemotherapy in patients with resected NSCLC. These data led several national and international groups to initiate a new generation of adjuvant trials in resected NSCLC, and more than 2,500 patients have already been included in these randomized studies. Chemotherapy can also be proposed preoperatively. Two randomized studies, which included 60 patients each, strongly suggested a benefit from neoadjuvant chemotherapy in patients with stage III NSCLC. A large French randomized study that included 375 patients with early stage NSCLC who did or did not receive preoperative chemotherapy has recently been completed. This study should help clarify the role of neoadjuvant chemotherapy in operable NSCLC. PMID- 9728588 TI - Gemcitabine and radiation therapy for non-small cell lung cancer. AB - Patients with stage III non-small cell lung cancer (NSCLC) frequently progress either within the irradiated field or systemically, due to uncontrolled microscopic dissemination present before the time of initial diagnosis. The use of combined modality therapy has led to improved survival rates in recent years. In particular, the use of cisplatin and vinblastine as induction chemotherapy is supported by two large randomized clinical trials. Nevertheless, the large majority of patients still die of progressive disease, thus providing a rationale for the integration of new active agents into the overall treatment plan of these patients. Gemcitabine has demonstrated significant single-agent activity in NSCLC. In addition, preclinical and early clinical data indicate that it is a powerful radiation enhancer. Clinical trials investigating this drug with concurrent radiation therapy in NSCLC are reviewed. PMID- 9728589 TI - Review of selected randomized trials in small cell lung cancer. AB - Approximately 25 years ago, investigators realized that small cell lung cancer was relatively sensitive to chemotherapy. The results of the initial trials showed that median survival duration in extensive-disease patients treated with combination chemotherapy was 8 to 10 months compared with 6 to 8 weeks for untreated patients. Similarly, in limited-disease patients, treatment with combination chemotherapy was associated with a median survival duration of approximately 12 months, while the median survival was approximately 3 to 4 months in patients treated with surgery or radiation therapy alone. Encouraged by the relatively high response rates and improved survival, investigators have conducted a large number of randomized trials that have tested concepts such as the Goldie-Coldman hypothesis and practical issues including chemotherapy dose and schedule, duration of treatment, and combined modality treatment. Selected randomized trials that have addressed important issues will be discussed in this brief review. PMID- 9728590 TI - Evaluation of new drugs in small cell lung cancer: the activity of gemcitabine. AB - The evaluation of the activity of new drugs against small cell lung cancer (SCLC) is discussed and the results with gemcitabine are presented. Gemcitabine has activity against untreated SCLC. Preliminary results from an ongoing study also indicate activity in so-called resistant SCLC. PMID- 9728591 TI - New agents in the management of advanced non-small cell lung cancer. AB - Non-small cell lung cancer accounts for 75% to 80% of the approximately 180,000 new cases of lung cancer that will develop in the Unites States this year. At the time of diagnosis, only 30% to 40% of patients are candidates for curative resection. Even for these patients, the 5-year survival rate ranges from 30% to 70%, indicating that many of these "early stage" patients have micrometastatic disease at the time of initial diagnosis. The remaining approximately 60% of newly diagnosed patients have either locally advanced unresectable disease or stage IV metastatic disease, and the majority of these patients will die of progressive disease before the first anniversary of their diagnosis. Overall, the 5-year survival rate for all newly diagnosed patients with non-small cell lung cancer has improved over the past two decades from 5% to 10% in the United States. PMID- 9728592 TI - Adjuvant chemotherapy for soft tissue sarcoma: does it make sense? PMID- 9728593 TI - New entities, issues, and controversies in the classification of malignant lymphoma. AB - The classification of malignant lymphoma has, for decades, provided a fertile area for controversy, discussion, and change. Such debate is necessary in order to appropriately assimilate new knowledge regarding this group of neoplasms. In this review, we provide a brief account of the evolution of classification schemes for lymphoma, and emphasize the recently proposed Revised European American Lymphoma (REAL) classification as a synthesis of current knowledge of clinical, morphologic, immunologic, and molecular data. Specific entities that are recently described or for which there are current controversies are discussed. The necessity of communication between clinicians and pathologists in the workup of a patient with malignant lymphoma is emphasized. PMID- 9728595 TI - Chromosomal abnormalities and molecular genetics of non-Hodgkin's lymphoma. AB - A number of recurring cytogenetic abnormalities have been identified in lymphomas that correlate with clinical, morphologic, and immunophenotypic features. For example, the t(14;18) is observed in a high proportion of follicular small cleaved cell lymphomas, most patients with translocations involving 3q27 have diffuse large cell lymphomas (B cell), and patients with a t(8;14) have either small noncleaved cell or diffuse large cell lymphomas. In contrast, a large proportion of neoplasms of T-cell origin are characterized by rearrangements that involve 14q11, 7q34-35, or 7p15. Molecular analysis of many of the recurring chromosomal translocations in lymphomas has resulted in the identification of the involved genes. Alterations in expression of these genes or in the production of an altered protein resulting from the rearrangement play an integral role in malignant transformation. In addition to chromosomal abnormalities, other types of mutations affecting oncogenes have been identified in lymphomas. This article reviews the genetic mutations involved in the pathogenesis of non-Hodgkin's lymphoma (NHL) with an emphasis on chromosomal abnormalities. PMID- 9728594 TI - B-cell development and maturation. AB - Approximately 85% of all non-Hodgkin's lymphomas arise from cells of the B lineage. Sequential stages of B-cell development have been defined by molecular markers, and these markers can be used to reclassify lymphoid malignancies as representing maturational arrest and immortalization at specific points in B-cell ontogeny. Several of the factors controlling the ordered rearrangement and expression of the immunoglobulin (Ig) genes have been identified. Signals generated by intermediates in Ig gene rearrangement, as well as by the complete Ig molecule, have been found to be critical in guiding early B-cell development. The processes of peripheral B-cell activation, antigenic affinity maturation, and terminal B-cell differentiation are also highly regulated. The molecular mechanisms responsible for both promoting and attenuating B-cell (humoral) immune responses have been increasingly well defined. This review summarizes some aspects of the current understanding of normal B-cell development, maturation, activation, and death, focusing on the factors implicated in regulating progression through these stages. PMID- 9728596 TI - The roles of human viruses in the pathogenesis of lymphoma. AB - There are two families of viruses that contribute to lymphomagenesis in humans: herpesviruses and retroviruses. The two herpesviruses are the Epstein-Barr virus (EBV) and human herpesvirus-8 (HHV-8). EBV is an extremely well-characterized transforming agent: nine viral proteins contribute to transformation in vitro. In contrast, in vivo, the pattern of EBV gene expression varies with different types of malignancies. EBV is associated with endemic Burkitt's lymphoma, acquired immune deficiency syndrome (AIDS)-related lymphoma, post-transplantation lymphoproliferative disease, Hodgkin's disease (HD), and rare T-cell lymphomas. We have summarized studies on the different patterns of viral gene expression and signaling in different EBV-related malignancies, which have begun to reveal how EBV variably contributes to the malignant phenotype in different diseases. HHV-8 is associated with primary effusion lymphomas in patients with AIDS, and the rapidly accumulating information on this virus is summarized. Human T-cell leukemia virus-1 (HTLV-1) is a retrovirus which is the causative agent of adult T cell leukemia/lymphoma (ATL). The specific mechanism of HTLV-1-mediated T-cell transformation is unclear, but the effects of HTLV-1 on interleukin-2 signaling are reviewed. PMID- 9728597 TI - Management of early-stage Hodgkin's disease: a continuing evolution. AB - The management of early-stage Hodgkin's disease (HD) has evolved during the past 25 years from exacting discovery of sites of disease using both clinical and surgical staging to treat with a specific radiation therapy field to determining disease parameters and treating with systemic chemotherapy. This evolution has occurred because of the increasing awareness of both early and late complications of radiation therapy, as well as because of data showing comparable results with combination chemotherapy without comparable morbidity. The final answer rests with ongoing, controlled clinical trials to determine whether chemotherapy alone will be the treatment of choice for early-stage HD. PMID- 9728598 TI - Current approaches to the management of non-Hodgkin's lymphoma. AB - Non-Hodgkin's lymphoma (NHL) is a complex malignancy with many different clinical presentations, histologies, and varied treatment options depending on the histologic type and other prognostic factors. This article focuses on the major histologic subtypes of NHL with discussion of the natural history of each and information on various treatment options. PMID- 9728599 TI - Lymphomas in the immunocompromised patient. AB - Lymphoma is a common opportunistic complication of immunosuppression. Lymphomas in patients with the acquired immunodeficiency syndrome (AIDS) may broadly be divided into four major types: intermediate- or high-grade systemic lymphoma, primary central nervous system (CNS) lymphoma, Hodgkin's disease (HD) and primary effusion lymphoma. Multiple active regimens have been identified for patients with AIDS-related systemic lymphoma. However, despite high initial complete response rates, most studies have reported a median survival of less than 1 year for these patients, with approximately half of the patients dying from lymphoma and half from opportunistic infections or other AIDS-related complications. The standard therapeutic approach for patients with AIDS-related primary CNS lymphoma is radiotherapy, although recent studies using combinations of chemotherapy with radiotherapy may offer an improvement in therapy for this group of patients who have very poor overall prognosis. Lymphoproliferative disease in patients after solid organ or bone marrow transplantation represents with a spectrum of disorders. No standard approach for therapy in this group of patients has been clearly established. PMID- 9728600 TI - The role of high-dose chemotherapy in patients with Hodgkin's disease and non Hodgkin's lymphoma. AB - Many patients with Hodgkin's and non-Hodgkin's lymphoma (NHL) can be cured today with combination chemotherapy and/or radiotherapy. However, for patients with suboptimal responses to initial therapy or for patients with refractory or relapsed disease, salvage therapy alone is usually inadequate to achieve long term survival. High-dose chemotherapy (HDC) with stem cell rescue has emerged as the treatment of choice for such patients as long-term disease-free survival can be obtained in a significant number of these patients. Dose-intensive treatment has been equivocally shown effective for certain patients with Hodgkin's and NHL, whether or not chemosensitivity is shown before transplant. However, HDC has yet to consistently yield durable responses in patients with indolent NHL. Additionally, perhaps the International Prognostic Index can now help identify "high-risk" NHL patients who may benefit from investigative approaches such as frontline HDC. PMID- 9728602 TI - Use of Rh immune globulin: ASCP practice parameter. American Society of Clinical Pathologists. AB - The use of Rh immune globulin (RhIG) has dramatically decreased the incidence of hemolytic disease of the fetus and newborn resulting from the production of anti D by an Rh-negative woman. However, despite the widespread use of RhIG, instances of Rh immunization continue to occur, most likely through failure to administer RhIG when indicated or in the appropriate dose. This utilization gap can be closed only through continued active surveillance by health care providers. The following report summarizes recommendations for the administration of RhIG, the dose required in various circumstances, prenatal and postnatal serologic testing of the obstetric patient, and the methods used to determine the degree of fetomaternal hemorrhage or the amount of Rh-positive RBCs in the circulation. PMID- 9728603 TI - Detection of Legionella by PCR in respiratory specimens using a commercially available kit. AB - Using polymerase chain reaction (PCR) for the detection of pathogens that are difficult to grow, such as Legionella species, may reduce difficulties encountered with culture and immunofluorescent staining. We evaluated a commercial PCR and hybridization kit, designed for environmental samples, for the detection of Legionella in respiratory specimens. Sixteen Legionella species cultures tested positive with the Perkin Elmer Legionella EnviroAmp Amplification and Detection kits (Perkin Elmer, Foster City, Calif). The assay detected as few as 100 colony-forming units per milliliter of spiked bronchoalveolar lavage (BAL) fluid, and no false-negative results were obtained. PCR inhibition by blood in the specimens was removed by washing pelleted specimens in sterile distilled water. Of 126 specimens screened with the kit, 1 induced sputum and 3 BAL specimens were positive by PCR. All 4 were validated as true-positive results by culture or serologic testing. The entire PCR and hybridization assay can be completed in less than 6 hours, whereas isolation and identification by culture requires up to 12 days, and serologic conversion may not be demonstrated for weeks. Molecular techniques based on direct extraction and amplification of DNA from respiratory specimens nay be useful for the timely diagnosis of legionellosis. PMID- 9728604 TI - Comprehensive evaluation of performance, laboratory application, and clinical usefulness of two direct amplification technologies for the detection of Mycobacterium tuberculosis complex. AB - The rapid detection of Mycobacterium tuberculosis from respiratory specimens is critical for optimal treatment of patients. Several nucleic acid amplification based systems designed to detect Mycobacterium tuberculosis complex directly from specimens have been developed, and 2 are commercially available. We studied the performance characteristics of these 2 systems (Gen-Probe Amplified Mycobacterium Tuberculosis Direct (MTD), Gen-Probe, San Diego, Calif; AMPLICOR, Roche Molecular Systems, Branchburg, NJ). Each uses a different amplification strategy, detection modality, and approach to inhibition of amplicon contamination. When compared with culture, the respective sensitivities and specificities were as follows: 92.2% and 98. 7% (study 1) and 88. 7% and 95.3% (study 2); AMPLICOR, 87.5% and 99.7%. Resolution of discordant results was accomplished by incorporating clinical data and multiple specimen analysis. An increased rate of false-positive results was encountered during 1 phase of the study. The conditions under which the test was performed were modified and the "contamination" issue was resolved. This report discusses the benefits and limitations of each assay, proposes cost effective algorithms for their incorporation into routine laboratory work flow, and discusses the clinical usefulness of these molecular technologies. PMID- 9728605 TI - The presence of CD34+ cell clusters predicts impending relapse in children with acute lymphoblastic leukemia receiving maintenance chemotherapy. AB - Early detection of relapse in children with acute lymphoblastic leukemia (ALL), as well as distinction of leukemic blasts from hematogones, can be difficult by morphologic examination alone. Using CD34 and terminal deoxynucleotidyl transferase (TdT) immunoperoxidase stains, we studied specimens from 25 children with ALL in morphologic remission to determine if we could identify children at risk of relapse. We studied morphologic remission bone marrow specimens from 9 patients who experienced relapse during the subsequent 6 months and 16 children who remained in complete remission, including 10 specimens with increased numbers of hematogones. Despite morphologic remission, clusters of more than 5 CD34+ and/or TdT-positive cells were identified before overt relapse in 6 of 9 cases of relapse, but were noted in only 1 of 10 specimens from children in continuous complete remission and none of 10 specimens with increased numbers of hematogones. Clusters of CD34+ or TdT-positive cells can identify individual patients at risk for imminent relapse. Hematogones may be differentiated from lymphoblasts by this method. PMID- 9728606 TI - Automated cell count in flow cytometry: a valuable tool to assess CD4 absolute levels in peripheral blood. AB - The enumeration of lymphocyte subsets in absolute counts has long relied on different methods applied separately to whole blood cell count, lymphocyte differential appreciation, and flow cytometric evaluation of lymphocyte subsets percentages. The development of multicolor labeling methods inflow cytometry now allows a more homogeneous appreciation of several cell subsets among gated lymphocytes. The use of internal calibrators, such as microbead suspensions, also permits a direct appreciation of subsets in absolute counts in a single-platform method. These methods were compared with a traditional multiplatform method of assessing absolute counts of lymphocyte subsets in a pilot study in which all manipulations were performed by 1 person and in a full-scale larger study performed in the normal working conditions of a hospital laboratory. Microspheres seem to be a reliable tool to perform absolute count enumeration inflow cytometry, but several precautions in the sample preparation and flow cytometric analysis are required. PMID- 9728608 TI - The TEST 1 automated system: a new method for measuring the erythrocyte sedimentation rate. AB - We evaluated performance of the TEST 1 (SIRE Analytical Systems, Udine, Italy), a fully automated analyzer for the measurement of the erythrocyte sedimentation rate (ESR). Intra-assay reproducibility was satisfactory for a wide range of ESR values, whereas there was a significant decrease in ESR data when the samples were stored at 4 degrees C for up to 24 hours. We compared TEST 1 with the Westergren ESR method approved by the International Council for Standardization in Haematology (ICSH) and with the Diesse Ves-Matic analyzer Linear regression analysis comparing the TEST 1 and the ICSH reference method yielded satisfactory correlations for K3 EDTA- and sodium citrate-anticoagulated samples. Bland-Altman analysis showed no evidence of a systematic bias between the TEST 1 and the reference method. A close correlation was found between the TEST 1 system and the Diesse Ves-Matic analyzer despite a significant positive systematic bias. Reference values for men and women were analyzed according to nonparametric statistics. The TEST 1 was easy to use, had a satisfactory operative practicability required minimal maintenance, and reduced contact with potential biohazards. This system enables the determination of ESR with any common standard sized tube; the use of samples anticoagulated with K3 EDTA can widely reduce the workload in clinical laboratories. PMID- 9728607 TI - Characterization of the lymphoid infiltrate in Hashimoto thyroiditis by immunohistochemistry and polymerase chain reaction for immunoglobulin heavy chain gene rearrangement. AB - A close relationship between Hashimoto thyroiditis (HT) and low-grade B-cell lymphoma of mucosa-associated lymphoid tissue (MALT) has been shown. We used immunohistochemistry to study paraffin sections from 40 unselected cases of HT and scored cases according to the lymphoid infiltrate and presence of lymphoepithelial lesions (LELs). Clonality was assessed by kappa/lambda immunohistochemistry and polymerase chain reaction for immunoglobulin heavy chain gene rearrangement (IgH PCR). Histologic findings were compared with 2 cases of primary thyroid MALT-type lymphoma. In HT, the lymphoid infiltrate consisted predominantly of T cells in all cases; B cells, associated with germinal centers, did not have the appearance of marginal zone cells. All cases had identifiable T cell LELs; immunohistochemistry confirmed inconspicuous, rare B-cell LELs in 13 of 40 cases. In all cases, plasma cells were polyclonal and IgH PCR showed a polyclonal pattern. Clinical follow-up was available for 34 patients. Lymphoma developed in none. In contrast, a B-cell predominant infiltrate of marginal zone cells was present in the MALT-type lymphomas that was not confined to germinal centers. Cytokeratin stains demonstrated severe loss of epithelial elements and destructive LELs. LELs are not, in isolation, a useful criterion for distinguishing low-grade MALT-type lymphoma of the thyroid from HT. Features associated with low-grade MALT-type lymphoma include a predominance of B cells, marked loss of epithelial elements, and destructive LELs composed of marginal zone B cells. Unselected cases of HT do not contain monoclones detectable by IgH PCR. PMID- 9728609 TI - Enhanced detection of malignant lymphoma in cerebrospinal fluid by multiparameter flow cytometry. AB - Immunophenotyping by flow cytometry has not been widely applied to cerebrospinal fluid (CSF) analysis. We attempted to optimize flow cytometric detection of malignant lymphoma in CSF samples by the routine use of 3- and 4-color flow cytometry, with specific selection of lymphoid cells by fluorescence vs 90 degrees light scatter gating. Thirty-six consecutive CSF samples were immunophenotyped by flow cytometry, and the results were compared with those of standard microscopic examination. Lymphoid events were adequate for analysis in 27 of the 36 samples. Each of the 9 unsuccessful samples was more than 24 hours old at analysis or contained fewer than 1 x 10(4) total cells (< or =1 cell/microL). Lymphoma was detected in 10 of the remaining 27 cases. Six lymphomas were detected by morphology and flow cytometry, 1 only by morphologic examination, and 3 only by flow cytometry. Therefore, the combination of flow cytometry and morphologic examination enhanced the detection by 43% over morphologic examination alone. Flow cytometry permitted the detection of lymphoid clones totaling less than 1% of total cells. Multicolor flow cytometry is a rapid and sensitive technique that enhances detection of lymphoma in paucicellular CSF samples. Given the great sensitivity of flow cytometry, future studies will be necessary to assess the significance of detecting small lymphoid clones in this setting. PMID- 9728610 TI - Detection of trisomy 3 in primary gastric B-cell lymphoma by using chromosome in situ hybridization on paraffin sections. AB - Recent studies in Western populations have shown that trisomy 3 is the most frequent chromosomal abnormality in primary gastric lymphoma (PGL). To study the incidence of trisomy 3 and its implications for the pathogenesis of PGL in Hong Kong, we have applied the technique of chromosome in situ hybridization in 13 cases of PGL by using archival paraffin-embedded tissue sections. Five cases of chronic gastritis were used as controls. Trisomy 3 was found in 9 (69%) of 13 cases, including cases of low-grade lymphoma and high-grade lymphoma with or without a low-grade component. Our findings are similar to the incidence of trisomy 3 reported in the Western literature. The persistent finding of trisomy 3 in various histologic grades of PGL suggests that it may be useful as a clonal marker in this group of neoplasms. Various molecular events involving chromosome 3 may be related to the pathogenesis of this group of lymphomas. PMID- 9728611 TI - Cancerization of lobules and atypical ductal hyperplasia adjacent to ductal carcinoma in situ of the breast. AB - Recurrent carcinoma develops in approximately 10% of patients with ductal carcinoma in situ (DCIS) of the breast treated with local excision and radiation therapy. Cancerization of lobules (COL) and atypical ductal hyperplasia (ADH)frequently occur at the edge of DCIS. We postulated that recurrent carcinoma is associated with ADH or COL near the DCIS excision margin, and the amount of DCIS left in the breast may be too large for eradication by radiation therapy. To identify histologic features associated with recurrence, we retrospectively studied specimens of 94 patients with DCIS treated by local excision and radiation. We analyzed the rim of tissue near the final margin for the amount of COL, ADH, and DCIS. During a median follow-up of 78 months, local recurrence developed in 9 patients. COL or ADH with DCIS near the final margin was associated with recurrence; the strongest relationship was with recurrences in the same site as the lumpectomy bed. DCIS with ADH was significantly associated with recurrence in the low-grade DCIS group; DCIS with COL was associated with recurrence in the high-grade group. Other features were not associated with outcome. We believe that ADH composed of cells identified as those of DCIS should be considered part of the DCIS lesion. DCIS may be inadequately excised if ADH and DCIS or COL and DCIS are near the margin. PMID- 9728612 TI - A comprehensive system to explore p53 mutations. AB - To establish an effective and reliable system for the detection of p53 mutations, we evaluated the detection efficiencies of nonisotopic polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP), fluorescence in situ hybridization (FISH), and immunohistochemistry. Ten cell lines (AsPc1, BxPc3, Miapaca2, Panc1, Colo320-011, Lovo, MCF7, LNCaP, HL-60, and Daudi), a peripheral blood sample from a patient with a p53 germline mutation (p53GML), and a normal peripheral blood sample were used for examination. Direct nucleotide sequencing identified p53 mutations in 7 of 12 samples (AsPc1, BxPc3, Miapaca2, Panc1, Colo320-011, HL-60, and p53GML). The nonisotopic PCR-SSCP detected anomalies of the PCR fragments in 5 cell lines. In the FISH analysis, 2 cell lines exhibited loss of heterozygosity of the p53 locus. Immunohistochemistry detected an accumulation of the abnormal p53 in 4 cell lines. The combination of these 3 methods produced no false-negative or false-positive results. This combination may be an excellent and beneficial system for the clinical diagnosis of the various human cancers. PMID- 9728613 TI - CD31 immunoreactivity in carcinomas and mesotheliomas. AB - CD31 is a specific and sensitive marker of endothelial differentiation. Previous reports have described its immunoreactivity in large series of soft tissue neoplasms, as well as its comparison with other available and commonly used endothelial markers. CD31 reactivity in carcinomas or mesotheliomas has been incompletely addressed, however. Hence, we applied anti-CD31 (JC70/A, DAKO, Carpinteria, Calif) to 290 previously characterized neoplasms by using a modified avidin-biotin-peroxidase complex technique following microwave epitope retrieval. Seven carcinomas showed plasmalemmal-based immunoreactivity (2 papillary thyroid carcinomas, 2 mucoepidermoid salivary gland carcinomas, 1 cutaneous adnexal tumor, 1 cutaneous squamous cell carcinoma, and 1 esophageal squamous cell carcinoma); the remaining 283 lesions were negative for this marker. We conclude that anti-CD31 immunostaining in carcinomas and mesotheliomas is rare. These findings support the concept that CD31 is a reliable marker of endothelial differentiation and should be included in diagnostic immunohistochemical panels when vascular tumors enter the differential diagnosis. PMID- 9728614 TI - Pathology of restenosis in saphenous bypass grafts after long-term stent implantation. AB - The implantation of saphenous vein grafts on the coronary arterial tree eventually leads to graft narrowing, which can be treated by the implantation of intravascular stents. However, long-term restenosis after stent implantation occurs in at least 30% of cases. Ten saphenous bypass grafts, in which a total of 12 stents had been implanted for an average of 32 months, were retrieved at least 10 months after implantation for angiographic diagnosis of reocclusion or severe restenosis. The metal struts were removed after macroscopic inspection of the vein, and the grafts were examined by light microscopy. Angiography revealed total occlusion in 9 stents and severe narrowing in 3. Pathologic examination revealed graft occlusion due to cellular hyperplasia in 4 cases and to recent thrombus formation in 5. Progression of atherosclerotic plaque was the cause of restenosis in the 3 severely narrowed grafts. In 2 of 5 grafts implanted with Palmaz-Schatz stents, the metallic struts had induced a local inflammatory reaction. Therefore, the long-term reocclusion of saphenous bypass grafts after stent implantation may be due to atherosclerotic plaque or fibromuscular hyperplasia. However, thrombus formation may still occur several years after implantation. In specific cases, stent implantation also induces inflammation around the stent struts. PMID- 9728615 TI - Thrombomodulin expression in transitional cell carcinoma. AB - Thrombomodulin (TM) is a surface glycoprotein reported to be expressed in a variety of tumors, including mesotheliomas, endothelial vascular tumors, squamous carcinomas, and various adenocarcinomas. This study evaluated TM expression in transitional cell carcinomas (TCCs) and determined whether immunostaining for TM has practical value in the diagnosis of TCCs. TM expression was observed in 96 of 106 primary tumors (bladder, 64/72; renal pelvis, 12/14; ureter, 3/3; prostate, 17/17) and in 21 of 23 metastatic TCCs. Among the adenocarcinomas, only 3 of 18 originating in the bladder, 7 of 46 in the lung, 4 of 21 in the breast, 2 of 24 in the ovary, and 2 of 4 in the pancreas expressed this marker. No staining was observed in the 22 renal cell carcinomas or the 35 adenocarcinomas of the prostate, 13 of the endometrium, or 12 of the colon. Nearly all squamous cell carcinomas (lung, 21/27; skin, 7/7; uterine cervix, 6/6; esophagus, 2/2; bladder, 2/2) reacted for TM. TM is a sensitive marker for TCC. TM immunostaining can assist in distinguishing this tumor from others, especially renal cell carcinomas and adenocarcinomas of the prostate, colon, and bladder, but it has no value in separating TCC from squamous cell carcinomas. PMID- 9728616 TI - The cost-effectiveness of routine histologic examination. AB - Although the histologic examination of routine tissues, such as hernia sacs and intervertebral disks, has shown a low incidence of detecting clinically significant unsuspected disease, the cost-effectiveness of histologic examination has not been determined. By using a theoretical model that assumed variable costs and gains in life expectancy secondary to detecting clinically significant disease, a threshold incidence of disease detection at which histologic examination is cost-effective was determined. By using the University of lowa (Iowa City) cost of examination (approximately $25), at least 1 of every 2,000 examinations would have to show clinically significant disease for histologic examination to be cost-effective. This threshold incidence decreases as production costs decrease or life-year values increase. Before definitive policy conclusions can be made, additional studies are needed to better define the trade off between cost and the value of information and the incidence of detecting clinically significant disease. PMID- 9728617 TI - Reactive mesothelial hyperplasia vs mesothelioma, including mesothelioma in situ: a brief review. AB - In biopsy tissue, discrimination between reactive mesothelial hyperplasia and epithelial mesothelioma can pose a major problem for the surgical pathologist. Confidence in the diagnosis is often proportional to the amount of tissue available for study and depends largely on findings of invasion and the extent and cytologic atypia of the lesion, because there is no marker specific for the mesothelium and that discriminates consistently among normal, hyperplastic, and neoplastic mesothelial tissue. Therefore, mesothelioma in situ is diagnosable only when invasive epithelial mesothelioma is demonstrable in the same specimen, in a follow-up biopsy specimen, or at autopsy. Comparison of 22 cases of mesothelioma in situ that fulfill these requirements for diagnosis with 141 invasive mesotheliomas and 78 reactive mesothelioses indicates that strong linear membrane-related labeling for epithelial membrane antigen and silver-labeled nucleolar organizer region-positive material that occupies 0.6677 microm2 or more of the nucleus in an atypical in situ mesothelial lesion of the pleura are found consistently in neoplastic mesothelial cells. Although these findings may engender suspicion of mesothelioma in situ in high-risk persons, the criteria for diagnosis of pure mesothelial lesions of this type are still under study. Mesothelioma in situ should be considered proved only when unequivocal invasion is identified in a different area of the pleura or at a different time; a diagnosis of pure mesothelioma in situ should not be made in patients not exposed to asbestos. PMID- 9728618 TI - Fine-needle aspiration biopsy in the management of thyroid nodules. PMID- 9728619 TI - Criteria for rebiopsy. PMID- 9728620 TI - The focus of "atypical glands, suspicious for malignancy" in prostatic needle biopsy specimens. PMID- 9728621 TI - France and United Kingdom channel efforts to improve health services. PMID- 9728623 TI - Social phobia--not just another name for shyness. PMID- 9728622 TI - Drug firm suit fails to halt publication of Canadian Health Technology Report. PMID- 9728624 TI - From the Centers for Disease Control and Prevention. Monitoring environmental disease--United States, 1997. PMID- 9728625 TI - From the Centers for Disease Control and Prevention. Epidemic malaria transmission--Armenia, 1997. PMID- 9728626 TI - A piece of my mind. The veteran. PMID- 9728627 TI - New options for the treatment of epilepsy. PMID- 9728628 TI - Acetaminophen and risk factors for excess anticoagulation with warfarin. PMID- 9728629 TI - Acetaminophen and risk factors for excess anticoagulation with warfarin. PMID- 9728630 TI - Acetaminophen and risk factors for excess anticoagulation with warfarin. PMID- 9728631 TI - Acetaminophen and risk factors for excess anticoagulation with warfarin. PMID- 9728632 TI - Acetaminophen and risk factors for excess anticoagulation with warfarin. PMID- 9728633 TI - Acetaminophen and risk factors for excess anticoagulation with warfarin. PMID- 9728634 TI - Internet health ratings systems: knowledge vs Babel. PMID- 9728635 TI - Internet health ratings systems: knowledge vs Babel. PMID- 9728636 TI - Internet health ratings systems: knowledge vs Babel. PMID- 9728637 TI - Internet health ratings systems: knowledge vs Babel. PMID- 9728638 TI - Mistreatment and maladaptations during medical internship. PMID- 9728639 TI - Mistreatment and maladaptations during medical internship. PMID- 9728640 TI - Mistreatment and maladaptations during medical internship. PMID- 9728641 TI - Effects of prenatal and postnatal methylmercury exposure from fish consumption on neurodevelopment: outcomes at 66 months of age in the Seychelles Child Development Study. AB - CONTEXT: Human neurodevelopmental consequences of exposure to methyl-mercury (MeHg) from eating fish remain a question of public health concern. OBJECTIVE: To study the association between MeHg exposure and the developmental outcomes of children in the Republic of Seychelles at 66 months of age. DESIGN: A prospective longitudinal cohort study. PARTICIPANTS: A total of 711 of 779 cohort mother child pairs initially enrolled in the Seychelles Child Development Study in 1989. SETTING: The Republic of Seychelles, an archipelago in the Indian Ocean where 85% of the population consumes ocean fish daily. MAIN OUTCOME MEASURES: Prenatal and postnatal MeHg exposure and 6 age-appropriate neurodevelopmental tests: the McCarthy Scales of Children's Abilities, the Preschool Language Scale, the Woodcock-Johnson Applied Problems and Letter and Word Recognition Tests of Achievement, the Bender Gestalt test, and the Child Behavior Checklist. RESULTS: The mean maternal hair total mercury level was 6.8 ppm and the mean child hair total mercury level at age 66 months was 6.5 ppm. No adverse outcomes at 66 months were associated with either prenatal or postnatal MeHg exposure. CONCLUSION: In the population studied, consumption of a diet high in ocean fish appears to pose no threat to developmental outcomes through 66 months of age. PMID- 9728642 TI - Paroxetine treatment of generalized social phobia (social anxiety disorder): a randomized controlled trial. AB - CONTEXT: The generalized type of social phobia (social anxiety disorder) is a severe and often disabling form of social anxiety that affects approximately 5% of the general population. Earlier research has shown monoamine oxidase inhibitors or benzodiazepines to be effective in treating this condition, but neither has achieved widespread use. OBJECTIVE: To compare the efficacy of paroxetine, a selective serotonin reuptake inhibitor, with placebo in adults with generalized social phobia. DESIGN: Twelve-week, multicenter, randomized, double blind trial. SETTING: Thirteen centers across the United States and 1 in Canada. PARTICIPANTS: Between April 13, 1995, and February 28, 1996, 187 persons meeting Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition criteria for generalized social phobia were randomized (and 183 returned for at least 1 efficacy assessment) to treatment. INTERVENTION: After a 1-week, single-blind, placebo, run-in period, patients received a double-blind, 11-week course of either paroxetine or matching-image placebo. The initial daily dosage of paroxetine (or placebo) was 20 mg with increases of 10 mg/d weekly (flexible dosing to a maximum of 50 mg/d) permitted after the second week of treatment. MAIN OUTCOME MEASURES: Number of responders based on the Clinical Global Impression Global Improvement Item ("much improved" or "very much improved"); mean change from baseline on the Liebowitz Social Anxiety Scale total score. RESULTS: Fifty (55.0%) of 91 persons taking paroxetine and 22 (23.9%) of 92 persons taking placebo were much improved or very much improved at the end of treatment (odds ratio [OR], 3.88; 95% confidence interval [CI], 2.81-5.36). Mean Liebowitz Social Anxiety Scale total scores were reduced by 39.1% (the mean baseline score of 78.0 declined by a mean of 30.5 points at follow-up) in the paroxetine group compared with 17.4% (the mean baseline score of 83.5 declined 14.5 points at follow-up) in the placebo group, a difference of 21.7% (95% CI, 8.7%-34.7%) favoring paroxetine. CONCLUSIONS: Paroxetine is an effective treatment for patients with generalized social phobia. Short-term (ie, 11-week) treatment results in substantial and clinically meaningful reductions in symptoms and disability. Future research should test whether these may be further reduced by extended treatment or supplementation with specific educational-cognitive behavioral techniques. PMID- 9728643 TI - Sunlight exposure and risk of lens opacities in a population-based study: the Salisbury Eye Evaluation project. AB - CONTEXT: Exposure to UV-B radiation in sunlight has been shown to increase the risk of cataract formation in high-risk occupational groups, but risk to the population has not been quantified. OBJECTIVES: To determine the ocular exposure to UV-B radiation in sunlight for a population of older persons and to determine the association between UV-B and lens opacities. DESIGN: The Salisbury Eye Evaluation project, a population-based cohort of older adults. SETTING: Salisbury, Md. PARTICIPANTS: A total of 2520 community-dwelling 65-year-old to 84 year-old adults in Salisbury, Md, from 1993 to 1995, of whom 26.4% were African Americans. MAIN OUTCOME MEASURE: Association of photographically documented cortical opacity 3/16 or greater in at least 1 eye with ocular UV-B exposure, reported in Maryland sun-years of exposure. RESULTS: The odds of cortical opacity increased with increasing ocular exposure to UV-B (odds ratio [OR], 1.10; 95% confidence interval [CI], 1.02-1.20). The relationship was similar for women (OR, 1.14; 95% CI, 1.00-1.30) and for African Americans (OR, 1.18; 95% CI, 1.04-1.33). Analyses of the ocular dose by each age group after the age of 30 years showed no vulnerable age group, suggesting damage is based on cumulative exposure. CONCLUSIONS: Although this population of older Americans has relatively low ocular exposure to UV-B in sunlight, there is still an association between ocular exposure and increasing odds of cortical opacity. Our study found an association among African Americans, which, to our knowledge, has not been reported previously. All sex and racial groups would benefit from simple methods to avoid ocular sun exposure. PMID- 9728644 TI - Lysophosphatidic acid as a potential biomarker for ovarian and other gynecologic cancers. AB - CONTEXT: Lysophosphatidic acid (LPA) has been shown to stimulate proliferation of ovarian cancer cells and is present in the ascitic fluid of patients with ovarian cancer. OBJECTIVES: To determine whether elevated levels of LPA are present in plasma from patients with ovarian cancer and other gynecologic malignancies compared with healthy controls and to evaluate whether an elevated LPA plasma level may be a biomarker for these diseases. DESIGN: A research assay was used to measure total LPA levels in plasma from healthy women and women with different diseases. All LPA assays and comparison of LPA levels and CA125 (an ovarian cancer biomarker) levels were performed by observers blinded to patient status or group. SETTING: The Cleveland Clinic Foundation. PARTICIPANTS: A convenience sample of 48 healthy control women, 48 women with ovarian cancer, 36 women with other gynecologic cancers, 17 women with benign gynecologic diseases, 11 women with breast cancer, and 5 women with leukemias. MAIN OUTCOME MEASURES: Total LPA levels in plasma samples from patients and controls. RESULTS: Patients in the ovarian cancer group had significantly higher plasma LPA levels (mean, 8.6 micromol/L; range, 1.0-43.1 micromol/L) compared with the healthy control group (mean, 0.6 micromol/L; range, <0.1-6.3 micromol/L) (P<.001). Elevated plasma LPA levels were detected in 9 of 10 patients with stage I ovarian cancer, 24 of 24 patients with stage II, III, and IV ovarian cancer, and 14 of 14 patients with recurrent ovarian cancer. Of 36 patients with other gynecologic cancers, 33 also showed higher LPA levels(mean, 14.9 micromol/L; range, <0.1-63.2 pmol/L), compared with healthy controls (P<.001). Elevated plasma LPA levels were detected in 5 of 48 controls and 4 of 17 patients with benign gynecologic diseases and in no women with breast cancer or leukemia. In comparison, among a subset of patients with ovarian cancer, 28 of 47 had elevated CA125 levels, including 2 of 9 patients with stage I disease. CONCLUSIONS: Plasma LPA levels may represent a potential biomarker for ovarian cancer and other gynecologic cancers. However, these findings are preliminary and require confirmation in larger studies. PMID- 9728645 TI - Late-term abortion. AB - Recent proposed federal legislation banning certain abortion procedures, particularly intact dilatation and extraction, would modify the US Criminal Code such that physicians performing these procedures would be liable for monetary and statutory damages. Clarification of medical procedures is important because some of the procedures used to induce abortion prior to viability are identical or similar to postviability procedures. This article reviews the scientific and medical information on late-term abortion and late-term abortion techniques and includes data on the prevalence of late-term abortion, abortion-related mortality and morbidity rates, and legal issues regarding fetal viability and the balance of maternal and fetal interests. According to enacted American Medical Association (AMA) policy, the use of appropriate medical terminology is critical in defining late-term abortion procedures, particularly intact dilatation and extraction, which is a variant of but distinct from dilatation and evacuation. The AMA recommends that the intact dilatation and extraction procedure not be used unless alternative procedures pose materially greater risk to the woman and that abortions not be performed in the third trimester except in cases of serious fetal anomalies incompatible with life. Major medical societies are urged to collaborate on clinical guidelines on late-term abortion techniques and circumstances that conform to standards of good medical practice. More research on the advantages and disadvantages of specific abortion procedures would help physicians make informed choices about specific abortion procedures. Expanded ongoing data surveillance systems estimating the prevalence of abortion are also needed. PMID- 9728646 TI - A 45-year-old man with low back pain and a numb left foot. PMID- 9728647 TI - A 28-year-old woman with multiple moles, 1 year later. PMID- 9728648 TI - Methylmercury exposure and neurotoxicity. PMID- 9728649 TI - Searching for a biomarker for ovarian cancer. PMID- 9728650 TI - JAMA, abortion, and editorial responsibility. PMID- 9728651 TI - Rationale for banning abortions late in pregnancy. PMID- 9728653 TI - JAMA patient page: food-borne illnesses. PMID- 9728652 TI - The continuing need for late abortions. PMID- 9728654 TI - Survival and DNA damage in Chinese hamster V79 cells exposed to alpha particles emitted by DNA-incorporated astatine-211. AB - Asynchronous Chinese hamster V79 lung fibroblasts were incubated at 37 degrees C for 30 min with the thymidine analog 5-[211At]astato-2'-deoxyuridine (211AtdU, exposure from DNA-incorporated activity) or with [211At]astatide (211At-, exposure from extracellular activity), and DNA-incorporated activity was determined. The 211AtdU content in cellular DNA increased as a function of extracellular concentration. Incorporation of 211At- was less than 1% of that of 211AtdU. After exposure, cells were frozen in the presence of 10% DMSO. One month later, survival was determined by the colony-forming assay, and DNA double-strand breaks (DSBs) were measured by the neutral elution method (pH 9.6). The survival curve for 211AtdU was biphasic (D37 = 2.8 decays per cell), reflecting killing of 211At-DNA-labeled cells and of unlabeled cells irradiated by 211At in neighboring labeled cells. The toxicity of 211At- decaying outside the cell (30-min exposure) was negligible. Analysis of the survival curve produced a D0 of 1.3 decays/cell for 211At-labeled cells. The yield of DSBs from the decay of DNA-incorporated 211At was compared with that from DNA-incorporated 125I. Each decay of 211At produced at least 10 times the number of DSBs as that obtained per 125I decay. The extreme radiotoxicity of DNA-incorporated 211AtdU seems to be associated with considerable damage to the mammalian cell genome. PMID- 9728655 TI - DNA fragmentation induced by a cytoplasmic extract from irradiated cells. AB - Apoptosis is a mode of cell death characterized by distinct morphological features and DNA fragmentation. The program that leads to apoptosis has been considered to be predominantly extranuclear, and a signal transduction pathway to the nucleus exists during apoptosis, while characteristic events occur in the nucleus. As for radiation-induced apoptosis, the signal transduction pathway remains unclear, especially the sites where the primary effect of radiation occurs. In this study, we demonstrate that a cytoplasmic extract prepared from irradiated cells has the ability to cause DNA fragmentation and that caspase-3 is activated in this extract. Normal nuclei of HeLa S3 cells were added to a cytoplasmic extract made from HL60 cells which had been irradiated with 30 Gy of 137Cs gamma rays and were incubated. Agarose gel electrophoresis of the added nuclei showed a characteristic DNA laddering pattern. This reaction was blocked by a caspase-3 inhibitor but not by an ICE inhibitor. These observations suggest that a signal transduction pathway from an unknown target of gamma radiation may exist upstream of caspase-3 during radiation-induced apoptosis. PMID- 9728656 TI - Increasing radiosensitivity in the course of fractionated X irradiation: the effect of contact with dead and dying cells. AB - We have measured survival after successive 2-Gy doses of X rays in HeLa cells and 1-Gy doses in cells of the nonimmortalized human fibroblast cell line AG15-22 under conditions where any effect of cell proliferation during multifraction X irradiation has been factored out. When HeLa cells in parallel series of (pseudo)hybrid spheroids (i.e. in agglomerates consisting of a mixture of supralethally irradiated HeLa feeder and viable HeLa cells) were exposed to n daily radiation doses and then trypsinized and exposed to the last dose, the surviving fraction at 2 Gy (SF2) declined exponentially from 0.55 +/- 0.01 to 0.31 +/- 0.01 after the fifth fraction. In monolayer HeLa cell cultures, the decline in SF2 was smaller but significant and was not influenced by the presence of feeder cells. Pure spheroids, composed entirely of viable HeLa cells, showed the same decline in SF2 as did monolayer cells. The cumulative-effect curve (i.e. the product of SF2 values) was linear-quadratic with the quadratic term increasing in the order monolayer, pure spheroids, pseudohybrid spheroids. SF2n and D0Eff (deduced from the initial SF2) severely underestimated cumulative radiosensitivity. This cumulative effect is clearly associated with the proximity of lethally irradiated cells and might be explained by differential population shifts in the course of the multifraction regimen. Similarly, AG15-22 cells irradiated with daily 1-Gy doses of X rays showed a larger increase in radiosensitivity when in hybrid spheroids than when in pure spheroids. However, for the AG15-22 cells, SF1 was twofold lower for the former than for the latter condition and remained constant for both conditions rather than decreasing with increasing fraction number. This large radiosensitizing effect remains unexplained. PMID- 9728657 TI - Paclitaxel-induced modification of the effects of radiation and alterations in the cell cycle in normal and tumor mammalian cells. AB - The cytotoxicity of paclitaxel (taxol) is associated mainly with block in G2/M phase, the most radiosensitive phase of the cell cycle. Nevertheless, taxol induced modification of the effects of radiation may vary from clear sensitization to subadditivity. Therefore, this effect was studied in relation to drug-induced alterations in the distribution of cells in the phases of the cell cycle in tumor cells (EMT-6 and OV-1063) and normal skin fibroblasts. Cell survival was evaluated with two colorimetric assays. The cell cycle was evaluated by FACS analysis of doubly-labeled cells. The radiosensitivity of the different cells studied was similar, apart from the less radiosensitive human fibroblasts. However, their dose- and time-dependent sensitivity to taxol varied significantly. After 24 h exposure of EMT-6 cells to taxol (IC50 approximately 20 nM), the fraction of cells in G2/M phase increased, the fraction in S phase decreased, and the proportion of possibly apoptotic cells with subdiploid and subtetraploid DNA content increased; this coincided with radiosensitization. In OV-1063 cells (IC50 approximately 3 nM), the drug-induced G2/M-phase block was most pronounced, but the combined effect with radiation was merely additive. In human fibroblasts (IC50 approximately 35 nM), a minimal G2/M-phase block with no change in the S phase and a massive elevation of apoptotic cells with subdiploid DNA content was accompanied by a subadditive combined effect with radiation. Six hours of exposure to taxol increased the fraction of cells in S phase in both nonsynchronized and S-phase-synchronized human fibroblasts (G1 phase approximately 65%, S phase approximately 13%). This was accompanied by a pronounced subadditive effect of the combined treatment. However, in G1-phase synchronized human fibroblasts (G1 phase > or =90%, S phase approximately 3%), only the fraction of cells in G2/M phase was slightly elevated, with a merely additive response to the combined treatment. The differences in the response to the combined treatment between slowly and rapidly proliferating cells in relation to modifications in the cell cycle are discussed. PMID- 9728658 TI - Glutathione ethylester protects against local and systemic suppression of contact hypersensitivity induced by ultraviolet B radiation in mice. AB - Irradiation of the skin with ultraviolet B (UVB) radiation causes a local and systemic suppression of T-cell-mediated immune responses. Recently, N acetylcysteine was found to protect against UVB-radiation-induced immunosuppression and several other types of damage induced by UV radiation. The protective effects appeared to be caused by an increase in glutathione (GSH). This increase was limited by feedback inhibition by GSH of its own synthesis. Better results were expected with the use of GSH derivatives which do not require de novo synthesis, such as GSH esters. In this study, topical application of glutathione ethylester (GSH-Et) was found to increase the epidermal GSH level in mice in a manner that was dependent on dose to 1234% of the control value at the highest dose tested (2.0 micromol/cm2). This resulted in dose-dependent protection against UVB-radiation-induced suppression of contact hypersensitivity. The highest dose of GSH-Et tested provided 83% protection against local suppression and 62% protection against systemic suppression. Immunosuppression induced by topically applied cis-urocanic acid (cis-UCA) was prevented completely. Although an effect on the formation of pyrimidine dimers cannot be excluded, the protective effect of GSH-Et seems to be mediated through inhibition of the action of cis-UCA. PMID- 9728659 TI - Effect of ionizing radiation on the release of cholecystokinin in the hypothalamus of the rat. AB - This study was designed to identify the mechanisms underlying the reduction in food intake in rats. Measurements were made of the release of cholecystokinin (CCK) stimulated by potassium chloride in the hypothalamus after (a) gamma irradiation (60Co), (b) treatment with the CCK-A and CCK-B antagonists L-364,718 and L-365,260 with and without radiation, (c) bilateral abdominal vagotomy, and (d) vagotomy with and without radiation and with and without L-364,718. The concentrations of CCK in hypothalamus perfusate were measured by a radioimmunoassay. Exposure of rats to 1, 3, 5 and 10 Gy (1 Gy/min) increased release of CCK in the hypothalamus in a manner that was dependent on dose. A dose of 5 Gy was chosen for further studies. Intraperitoneal (i.p.) administration of 10, 20 and 50 microg/kg of L-364,718 did not induce significant changes in release of CCK in sham-irradiated animals. However, the drug decreased the release of CCK induced by radiation in a dose-dependent manner. In contrast to L 364,718, 20-50 microg/kg of L-365,260 decreased the release of CCK in the hypothalamus in sham-irradiated animals but did not decrease release of CCK induced by exposure to radiation. Vagotomy produced an insignificant reduction in release of CCK compared to that in sham-irradiated controls. However, vagotomy decreased release of CCK in irradiated rats compared to the irradiated rats without vagotomy. Vagotomy and i.p. administration of 10, 20 and 50 microg/kg of L-364,718 decreased release of CCK in irradiated rats compared to that in irradiated rats without vagotomy. However, i.p. administration of 10, 20 and 50 microg/kg of L-364,718 did not induce significant decreases in release of CCK in the hypothalamus of vagotomized and irradiated animals compared to those in rats that were vagotomized and irradiated but not treated with L-364,718. These results demonstrate that radiation increases the release of CCK in the hypothalamus, and that this effect is inhibited by vagotomy and the administration of a CCK-A receptor antagonist. A CCK-A receptor antagonist may be used to mitigate a radiation-induced deficit in food intake. PMID- 9728660 TI - Histopathological and morphometric study of the late effects of heavy-ion irradiation on the spinal cord of the rat. AB - The late effects of heavy-ion irradiation on the spinal cord of the rat were investigated histologically and morphometrically. After a single exposure of each animal's lower thoracic and lumbar spinal cord to a carbon-ion beam, the animals were observed clinically for up to 69 weeks and their spinal cords were examined histologically after sacrifice. Paralysis of the hind limbs appeared from 16 to 20 weeks after irradiation with 20 Gy or more. The first histological change seen was vacuolization in the marginal white matter, which appeared 19 to 25 weeks after irradiation with more than 10 Gy. After irradiation with more than 15 Gy, bilateral destructive cavities occurred in the white matter, especially in the lateral tract. These histological changes were similar to those reported frequently for X irradiation. The mean cross-sectional area of the blood vessels in the irradiated spinal cord increased in a manner that was dependent on dose and was significantly larger 15 to 17 weeks after irradiation with 30 Gy. Reconstruction of small destructive lesions from serial sections consistently revealed dilated veins in the centers of these lesions. The effective dose that induces 50% incidence of hind-limb paralysis and destructive cavity formation (ED50) as determined using a curve-fitting method was 18.5 and 19.5 Gy, respectively, and the latent period was shorter than that for X irradiation. PMID- 9728661 TI - Mortality in beagles irradiated during prenatal and postnatal development. I. Contribution of non-neoplastic diseases. AB - To evaluate the lifetime health effects of exposure to ionizing radiation during development, 1,680 beagles received whole-body exposures to 60Co gamma rays or sham exposures. Eight groups of 120 dogs each received mean doses of 15.6-17.5 or 80.8-88.3 cGy in early, mid- or late gestation, at 8, 28 or 55 days after breeding, or at 2 days after birth. Another group of 120 dogs received a mean dose of 82.6 cGy as 70-day-old juveniles and one group of 240 dogs received a mean dose of 81.2 cGy as 365-day-old young adults. Sham irradiations were given to 360 controls. Sexes were equally represented. There was no significant effect of irradiation on mean survival times in any groups. In 1,343 dogs allowed to live out their life span, chronic renal disease was a common cause of mortality, and irradiation in the late fetal or juvenile periods potentiated this disease, resulting in increased mortality due to renal failure. This was consistent with earlier findings of the high radiosensitivity of the kidney in the perinatal period. Hypothyroidism associated with atrophic thyroiditis was decreased by irradiation, a finding contrary to expectation and not easily explained. Diabetes mellitus was increased by irradiation in the mid- and late gestation and juvenile periods, a finding which is intriguing based on early reports of a similar finding in atomic bomb survivors. Though convulsive seizures were a common cause of mortality in the dogs, there was no evidence for increased risk associated with prenatal irradiation as has been reported in humans. Genetic analyses indicated that renal disease, hypothyroidism, diabetes mellitus and convulsive seizures all had a heritable component, but that this did not influence or bias the radiation responses evaluated. PMID- 9728662 TI - Mortality in beagles irradiated during prenatal and postnatal development. II. Contribution of benign and malignant neoplasia. AB - To evaluate the lifetime carcinogenic hazards of exposure to ionizing radiation during development, 1,680 beagles received whole-body exposures to 60Co gamma rays or sham exposures. Eight groups of 120 dogs each received mean doses of 15.6 17.5 or 80.8-88.3 cGy in early, mid- or late gestation, at 8, 28 or 55 days postcoitus or at 2 days after birth. Another group of 120 dogs received a mean dose of 82.6 cGy as 70-day-old juveniles and one group of 240 dogs received a mean dose of 81.2 cGy as 365-day-old young adults. Sham irradiations were given to 360 controls. Sexes were equally represented. In 1,343 dogs allowed to live out their life span, neoplasia was a major disease, contributing to mortality in 40% of the dogs. There was a significant increase in benign and malignant neoplasms occurring in young dogs (<4 years old), including fatal malignancies, after irradiation in the perinatal (late fetal and neonatal) periods. The lifetime incidence of fatal neoplasms was also increased in dogs irradiated perinatally. Three malignancies-lymphomas, hemangiosarcomas and mammary carcinomas-accounted for 51% of all fatal tumors. There was an apparent lifetime increase and earlier onset of lymphomas in dogs exposed as fetuses. Fatal hemangiosarcomas were increased in dogs irradiated early and late in gestation. Fatal mammary carcinomas were not increased by irradiation, although non-fatal carcinomas were increased after perinatal exposure. Myeloproliferative disorders and central nervous system astrocytomas appeared to be increased in perinatally irradiated dogs. These data suggest that irradiation in both the fetal and neonatal periods is associated with increased early onset and lifetime cancer risk. PMID- 9728663 TI - Chernobyl-related thyroid cancer in children of Belarus: a case-control study. AB - The accident at the Chernobyl nuclear power plant on April 26, 1986, released approximately 2 EBq of 131I and other radioiodine isotopes that heavily contaminated southern Belarus. An increase in thyroid cancer reported in 1992 and attributed to the Chernobyl accident was challenged as possibly the result of intensive screening. We began a case-control study to test the hypothesis that the Chernobyl accident caused the increase in thyroid cancer. Records of childhood thyroid cancer in the national therapy centers in Minsk in 1992 yielded 107 individuals with confirmed pathology diagnoses and available for interview. Pathways to diagnosis were (1) routine endocrinological screening in 63, (2) presentation with enlarged or nodular thyroid in 25 and (3) an incidental finding in 19. Two sets of controls were chosen, one matched on pathway to diagnosis, the other representing the area of heavy fallout, both matched on age, sex and rural/urban residence in 1986. The 131I dose to the thyroid was estimated from ground deposition of 137Cs, ground deposition of 131I, a data bank of 1986 thyroid radiation measurements, questionnaires and interviews. Highly significant differences were observed between cases and controls (both sets) with respect to dose. The differences persisted within pathway to diagnosis, gender, age and year of diagnosis, and level of iodine in the soil, and were most marked in the southern portion of the Gomel region. The case-control comparisons indicate a strong relationship between thyroid cancer and estimated radiation dose from the Chernobyl accident. PMID- 9728665 TI - Response of the canine esophagus to irradiation. AB - One hundred twenty-eight beagle dogs were randomized to receive thoracic irradiation with doses between 0 and 72 Gy in 1.5-Gy fractions over 6 weeks. Dogs were randomized to have either 33, 67 or 100% of their lung volume irradiated. The entire thoracic portion of the esophagus and variable portions of the fundus of the stomach were included in the treatment field at all volumes. Sixteen of the 128 dogs entered in the study developed clinical signs of esophagitis. These 16 dogs received doses between 45 and 72 Gy. Clinical signs of esophagitis/gastritis included dysphagia, anorexia, emesis, excessive salivation and weight loss that required force-feeding of a liquid diet. An ED50 of 67.2 Gy (95% CI 61.45-79.7 Gy) was calculated for the occurrence of clinical signs that required some supportive treatment. Three of the 16 dogs receiving 63 or 72 Gy failed to respond to treatment and were euthanized. Twenty-five other dogs were euthanized prior to 2 years due to other treatment-related complications. Two dogs died of causes not related to treatment. No late esophageal complications were observed in the remaining 98 dogs out to 2 years after irradiation. Esophageal specimens from 79 dogs were available for quantitative histological analysis 2 years after irradiation. Histological analysis showed a decrease in the percentage of glandular tissue with a corresponding increase in lamina propria and muscle. PMID- 9728664 TI - Intracranial tumors after exposure to ionizing radiation during infancy: a pooled analysis of two Swedish cohorts of 28,008 infants with skin hemangioma. AB - The risk of intracranial tumors after exposure to ionizing radiation during infancy has been studied in a pooled analysis of two Swedish hemangioma cohorts (n = 28,008). The mean absorbed intracranial dose was low (7 cGy, range 0-11.5 Gy). The cohorts were followed up in the Swedish Cancer Register for incident intracranial tumors during the period 1958-1993. Eighty-eight tumors were found in 86 individuals compared to 60.72 expected [standardized incidence ratio (SIR) 1.42, 95% confidence interval (CI) 1.13-1.75]. The SIR increased significantly in ascending dose categories (P = 0.02). Dose-response analyses were performed with Poisson regression methods. There was a significant effect of dose, and the dose effect relationship was negatively modified by age at first treatment. This indicates a higher risk for those exposed earlier in life. A linear dose-response model modified by age at first treatment resulted in the best fit. The excess relative risk (ERR) was 2.7/Gy (95% CI 1.0-5.6). The ERR/Gy was 4.5 if the treatment was given before 5 months of age, 1.5 if it was given at 5-7 months and 0.4 if it was given later. The study thus strongly indicates that there exists a dose-response relationship between absorbed dose in the brain and the subsequent risk of developing an intracranial tumor and that the risk is higher among infants exposed at younger ages. PMID- 9728666 TI - Partition of thorium between organs of monkeys injected with thorotrast: implications for alpha-particle dosimetry. AB - Risk estimates for alpha-particle-induced malignancies have been based mainly on studies of Thorotrast patients, but certain aspects of its deposition in the body have been at issue: the partition between the liver, spleen and red bone marrow, and the deposition at lower concentrations in other organs, such as muscle and fat, which may contribute to the risk. To supplement the existing data for humans, thorium concentrations were measured in the organs of two female monkeys 3-4 years after injection with Thorotrast. Relative deposits (liver:spleen:red bone marrow) were 54:6:41 and 75:4:21, in better agreement with the most recent observations in Thorotrast patients than with previous reports. Whereas the human testis had ranked among intermediate-level organs such as the adrenal glands and pancreas, the ovary of the monkey was among the organs with the lowest concentrations. The data suggest that risk factors for induction of malignancies by alpha-particle irradiation should be re-examined. PMID- 9728667 TI - Sexual problems in healthy and depressed persons. AB - In the ongoing Zurich Cohort Study, 591 males and females from the general population of Zurich were interviewed five times over the 15-year period between the ages of 20 and 35 years. The data on sexual problems were mainly obtained from the final three interviews when the subjects were aged 28-35 years. Emotional problems or sexual dysfunction alone are rare and are almost always associated with each other, or with changes in libido. However, changes in libido without associated emotional problems or sexual dysfunction are relatively common. Libido therefore appears to be the core problem with which other sexual problems overlap. Overall, sexual problems of some type were found in 26% of normal subjects, 45% of non-treated depressed patients and 63% of treated depressed patients. This increase in sexual problems in treated depressed patients is mainly due to an increase in sexual dysfunction and emotional problems; the level of libido appears not to be affected by treatment. There was no difference in the prevalence of sexual problems of any kind between patients treated with medication and those treated only with psychotherapy. PMID- 9728668 TI - The behavioural toxicity of antidepressants: effects on cognition and sexual function. AB - The cognitive system is structured from sets of schema, patterns of neural activity that allow the assimilation or accommodation of new experiences and so, by a process of consolidation, the gradual development of knowledge and understanding. As well as schema for purely cognitive processes, there are similar structures that enable individuals to deal with sexual behaviour and affectual relationships (e.g. hedonia, self-esteem, personal preferences and body image). In depression, there is a well established disruption of cognitive function that results in anhedonia and a loss of pleasure, including that from sexual activities. Many antidepressants also have a direct pharmacological action on the central nervous system and disrupt cognitive function, so increasing anhedonia and impairing sexual function. Drug actions on cognitive structures, which in turn increase anhedonia and reduce sexual libido, are over and above any direct pharmacological effects on the more overt behavioural activities associated with sex, including orgasm, erectile function, potency and ejaculation. The tricyclic antidepressants, for example, destroy the cognitive structures that are vital to maintain normal libido as well as disturbing overt sexual behaviours. Some selective serotonin reuptake inhibitors (SSRIs; paroxetine and sertraline) are associated with behavioural activation that is also responsible for an impairment of sexual function. However, there are clear differences between the SSRIs, and fluvoxamine (relative to the other SSRIs) has little effect on objective measures of cognition or on cerebral and behavioural components of sexual function. PMID- 9728669 TI - Serotonin, serotonergic receptors, selective serotonin reuptake inhibitors and sexual behaviour. AB - The serotonergic system in the brain modulates many types of behavioural and physiological processes. An example of this modulatory function is seen with the selective serotonin reuptake inhibitors (SSRIs) which enhance serotonin transmission and influence mood, anxiety states, aggression, feeding and sexual behaviour. At present, 14 different serotonin receptors have been described and, although the function and localization of many of these receptors is becoming increasingly clear, much remains unknown. The SSRIs are intriguing drugs; by blocking presynaptic and somatodendritic serotonin transporters, they enhance serotonergic neurotransmission and thereby activate serotonin receptors. It is this effect which leads to the characteristic effects of the SSRIs. Theoretically, however, it appears possible that they may have differential effects on the various subpopulations of serotonin receptors. Differences between the SSRIs have recently been reported in males with rapid ejaculation; fluvoxamine, in contrast to other SSRIs, did not affect rapid ejaculation. What difference in the mechanism of action between the SSRIs is responsible for this differential profile? A conditioned taste aversion procedure has been used in mice to investigate the discriminatory stimuli (cues) of fluvoxamine and fluoxetine. It appeared that the discriminatory stimulus of fluvoxamine is primarily mediated via 5-hydroxytryptamine (HT)1A receptors, whilst that of fluoxetine is primarily mediated via 5-HT2C receptors. Both types of receptors have been implicated in depression and it is conceivable that different SSRIs have intrinsic activity at these receptors. Investigations are now ongoing to determine whether this differential mechanism of action also applies to the other SSRIs and whether there are differences between the SSRIs with respect to their effect on sexual behaviour in rodents. PMID- 9728670 TI - The biological basis of female sexuality. AB - Human sexuality has three main roots: biological, motivational-affective relational, and cognitive. Unfortunately, in women, the biological dimension is usually disregarded. Hormones are necessary, but not sufficient, factors to maintain a satisfying human libido. In women, oestrogens prime the central nervous system, acting as neurotrophic and psychotrophic factors throughout life. They also prime the sensory organs, including the skin with its sebaceous and sweat glands, which are the key receptors for external sexual stimuli. Oestrogens are also the 'permitting factors' for the action of vaso-intestinal peptide, the key neurotransmitter involved in the endothelial and vascular changes leading to vaginal lubrication. Other factors, such as medication, alcohol and other health problems, can modify the biological impact of hormones on libido. Depression may cause a progressive decline in interest in sexual behaviour leading to low libido, difficulty in sexual arousal, secondary anorgasmia and/or frank sexual aversion. Increasing attention of doctors towards the sexual problems of women will dramatically improve female quality of life, especially during difficult periods of transition. PMID- 9728671 TI - Depression: a long-term illness and its treatment. AB - Depression is a common illness that is frequently chronic or recurring. It is associated with substantial disability and therefore treatment strategies need to take into account its long-term course. The risk of relapse can be reduced provided therapy is adequate, in terms of both duration and dose, and there is now good evidence for some antidepressants that long-term treatment also reduces the risk of recurrence (i.e. new episodes of depression). This aspect of efficacy has been investigated most thoroughly with the tricyclic antidepressant imipramine and with the selective serotonin reuptake inhibitors. The clearest demonstration of the ability to reduce the risk of new episodes of depression is obtained from studies designed to test prophylaxis specifically in patients who have responded to antidepressant treatment and who have maintained their response during a period of continuation treatment to ensure resolution of the episode. The long-term efficacy of imipramine and fluoxetine was demonstrated using this design. More recently fluvoxamine was shown to be effective in reducing the risk of new episodes in a 1-year study in patients whose acute episode of depression had responded to treatment with fluvoxamine and who had remained well for 18 weeks. These prophylactic studies show that antidepressants reduce the risk of new episodes of depression, and prophylactic treatment should therefore be continued as long as the risk persists. PMID- 9728672 TI - Selective serotonin reuptake inhibitor-induced sexual dysfunction: clinical and research considerations. AB - Antidepressants, including the tricyclic antidepressants, monoamine oxidase inhibitors (MAOIs) and selective serotonin reuptake inhibitors (SSRIs), cause sexual dysfunctions such as decreased sexual desire, erectile difficulties and delayed ejaculation. Such sexual side-effects affect quality of life and may result in non-compliance with medication and the associated risk of recurrence of depression. Depression may also be associated with sexual disturbances, especially reduced libido. It is important to unravel the origin of sexual problems during depression and determine whether they were present before depression started, whether they are associated with the depression, or whether they are an effect of medication. Baseline measurements, objective measures and accurate instruments are all essential for scientific research into the sexual side-effects of antidepressants. Various human factors that may influence measurements of sexual behaviour must also be taken into account. Considering all these provisos, the ejaculation delaying effects of the SSRIs (fluvoxamine, fluoxetine, paroxetine and sertraline) have been investigated in a double-blind, placebo-controlled study in men with rapid ejaculation. The SSRIs were given at their recommended daily dosages for 6 weeks and the men measured their intravaginal ejaculation latency time at home using a stopwatch. The results showed a clear difference between the SSRIs, fluvoxamine having by far the least disturbing effect on ejaculation. PMID- 9728673 TI - Psychotropics and sexuality. AB - Relatively little is known about the sexual side-effects of psychotropic drugs, probably due to the taboos surrounding discussion of sexual matters. However, there is a growing interest in this topic. The antidepressants are probably the most widely studied class of psychotropic drug and evidence suggests that all commonly used classes are associated with some sexual side-effects. However, there may well be differences between the effects of different drugs in the same class. The study of sexual side-effects in bipolar patients taking mood stabilizers is complicated by the existence of numerous confounding variables and only limited data are currently available on the sexual side-effects of benzodiazepines and antipsychotics. It is also important to take into account a number of methodological considerations when interpreting data on the sexual side effects of psychotropic drugs. PMID- 9728674 TI - Intraventricular pressure drop and aortic blood acceleration as indices of cardiac inotropy: a comparison with the first derivative of aortic pressure based on computer fluid dynamics. AB - This paper presents a computational approach to ventricular fluid mechanics to evaluate three inotropic indices of early ejection: the intraventricular pressure drop (deltap). the first derivative of aortic flow rate (df/dt) and the first derivative of aortic pressure dp/dt. dp/dt is one of the most frequently used indices for assessing myocardial inotropy. Deltap and df/dt are characteristic of inertia driven flows and reflect the impulsive nature of the flow inside the ventricle during the ejection phase. The study is based on an axisymmetric fluid dynamics model of the left ventricle, developed according to the finite element approach. The fluid cavity is bounded by a shell containing two sets of counter rotating contractile fibres. Two simulation sets were performed: the former to investigate the sensitivity of deltap and df/dt peaks (deltap(max) and df/dt(max)) with respect to changes in the inotropic state of the fibre. The latter allows the evaluation of the dependency of deltap(max) and df/dt(max) on afterload by means of two supravalvular stenoses of 50% and 70%. The model simulates the inertial features of ventricle behaviour. The calculated values of the indices investigated are in close agreement with those reported in the literature. The sensitivities of deltap(max) df/dt(max) and dp/dt(max) are calculated for the two simulation sets. Data are normalised with respect to the maximum values reached in the simulation set. The comparison indicates that deltap(max) has a greater sensitivity (3.4 vs. 3.1 ) and a more linear pattern than dp/dt(max) for changes in the inotropic state of the fibre. df/dt(max), shows a sensitivity close to dp/dt(max). Results confirm that the afterload does not affect dp/dt(max), in accordance with experimental observations, while deltap(max) and, to a major degree, df/dt(max) decrease when the afterload is increased. PMID- 9728675 TI - Pressure variation under the ischial tuberosity during a push cycle. AB - The present study is devoted to the variation of the magnitude of the compressive loading acting on the soft seating parts of a disabled person and the related pressure distribution under the ischial tuberosity during wheelchair propulsion. A combined experimental and computational approach was designed to predict correctly the change in magnitude of the maximum internal shear and compressive stresses produced by different propulsion speeds, cushion characteristics and body position of the subject. The results obtained show that the vertical force acting on the seating parts increases with the propulsion speed and exceeds the body weight by more than 100%. The related pressure under the ischial tuberosity shows a significant increase of 125% on the tissue/seat interface and an estimated increase of 185% in the peak compressive stress. It is concluded that computer modelling using a quasi-static approach provides a reliable estimate of the pressure values by the observed loading frequencies of 0-4 Hz. It can also be noted that the time independent material model utilised for the bulky soft tissue proved adequate for the estimate of the pressure level occurring under the ischial tuberosity during a push cycle. PMID- 9728676 TI - Codman's paradox of the arm rotations is not a paradox: mathematical validation. AB - Movement of a straight arm centred at the shoulder joint in three successive 90 degrees rotations, each around the respective orthogonal coordinate axis, leads to an apparently unrelated 90 degrees rotation around the longitudinal arm axis. This empirical fact is known as Codman's paradox, after a Bostonian surgeon who first reported it in 1934. However, by means of homogeneous coordinates, it is herein demonstrated that the phenomenon is just a mechanical property mathematically described by the equivalence between the matricial product of three orthogonal rotation matrices applied to a position vector and the matricial product of a single rotation matrix applied to the same vector. The latter rotation matrix corresponds to the middle one in the former group of three. When polar coordinates are used, the demonstration is even simpler, for the total shift vector clearly shows a single net effect on the longitudinal axis rotation. Thus, Codman's paradox is not a paradox. This property improves the muscle dynamics arm knowledge and might find applications in robotics. PMID- 9728677 TI - Local identifiability for two and three-compartment pharmacokinetic models with time-lags. AB - In this paper, we show that time-lags between compartments in a 2 and 3 compartment pharmacokinetic model may be taken into account but that separate identification for model parameters and for time-lags would not be suitable. Furthermore, it may happen that a time-lag model is locally identifiable while the corresponding model without delay is not. For two-compartment delayed models, with only one observation, it is not necessary to have two different inputs contrary to the case without time-lag. Both the Laplace transformation and a Jacobian matrix are used in an identifiability study. For all two-compartment models we have investigated which kind of parameters or lags are identifiable from amount (Q) or concentration (C) measures. PMID- 9728678 TI - Three-dimensional finite element analysis of thermal shock in a premolar with a composite resin MOD restoration. AB - A three-dimensional finite element model of a human premolar with a composite resin Class II MOD has been used to investigate the temperature changes and induced thermal stresses associated with the imbibing of a hot liquid. Regions of high tensile stress were revealed and their possible clinical significance with respect to microleakage and failure in the approximal region of the restoration are discussed. PMID- 9728679 TI - Three-dimensional dynamic behaviour of the human knee joint under impact loading. AB - The objective of this study is to determine the three-dimensional dynamic response of the human knee joint. A three-dimensional anatomical dynamic model was thus developed and consists of two body segments in contact (the femur and tibia) executing a general three-dimensional dynamic motion within the constraints of the different ligamentous structures. Each of the articular surfaces at the tibio-femoral joint was represented mathematically by a separate mathematical function. The joint ligaments were modelled as nonlinear elastic springs. The six-degrees-of-freedom joint motions were characterized by using six kinematic parameters, and ligamentous forces were expressed in terms of these six parameters. Knee response was studied by considering sudden external forcing pulse loads applied to the tibia. Model equations consist of nonlinear second order ordinary differential equations coupled with nonlinear algebraic constraint conditions. Constraint equations were written to maintain at least one-point contact throughout motion; one- and two-point contact versions of the model were developed. This Differential-Algebraic Equations (DAE) system was solved by employing a DAE solver: the Differential/Algebraic System Solver (DASSL) developed at Lawrence Livermore National Laboratory. A solution representing the response of this three-dimensional dynamic system was thus obtained for the first time. Earlier attempts to determine the system's response were unsuccessful owing to the inherent numerical instabilities in the system and the limitations of the solution techniques. Under the conditions tested, evidence of "femoral roll back" on both medial and lateral tibial plateaus was not observed from the model predictions. In the range of 20 degrees to 66 degrees of knee flexion, the lateral tibial contact point moved posteriorly while the medial tibial contact point moved anteriorly. In the range of 66 degrees to 90 degrees of knee flexion, contact was maintained only on the medial side and the tibial contact point (on the medial side) continued to move anteriorly. It was further found that increasing pulse amplitude and/or duration caused a decrease in the magnitude of the tibio-femoral contact force at a given flexion angle. These results suggest that increasing load level caused a decrease in joint stiffness. The results of this study also show that the anterior fibres of the posterior cruciate and the medial collateral ligaments are the primary restraints for a posterior forcing pulse in the range of 20 degrees to 90 degrees of knee flexion; this explains why most isolated posterior cruciate ligament injuries and combined injuries to the posterior cruciate and the medial collateral result from a posterior impact on a flexed knee. PMID- 9728680 TI - A fast responding combined direct and indirect calorimeter for human subjects. AB - The construction and performance of a 5.4 m3 combined direct and indirect calorimeter for human subjects is described. The calorimeter was constructed for studies on human subjects primarily undergoing fast alterations in heat production and heat losses, e.g. after a meal or during physical exercise. A heat sink and a heat substitution principle is used to measure sensible heat losses directly. Evaporative heat losses are determined by measuring water vapour input and output. Indirect calorimetry is performed by measuring output air flow and changes in gas composition of the air entering and leaving the calorimeter. The response times (90%) for sensible heat and evaporative heat were found to be 4 min and 34 min, respectively. Three alcohol combustion tests gave a recovery of CO2 and O2 in the range 92-97%. The recovery of water was found to be in the range 56-89%. PMID- 9728681 TI - Kinematics of the knee joint in deep flexion: a radiographic assessment. AB - The purpose of this study is to describe the kinematics of normal knees in vivo, assessed in deep flexion, using bi-planar radiographs. Antero-posterior and lateral views were obtained from five healthy males during three sequential positions of kneeling. In the first position, the subject knelt with the knees fully flexed (deep flexion between 150 and 165 degrees) and torso upright. In the second position, the subject bowed forward to an intermediate position (about 120 degrees of knee flexion). In the third position, the subject bowed further until his head touched the floor, supporting the upper torso with hands and with the knees flexed at about 90 degrees. The results show that past 135 degrees of knee flexion, the patella cleared the femoral groove and was in contact only with the condyles. For these particular postures, and during deep flexion, motion of the femur on the tibia did not reveal the classical femoral 'roll back'. Rather the lateral femoral condyle rolled further over the postero medial aspect of the lateral tibial plateau while contact of the medial femoral condyle occurred more anteriorly, but still in the posterior part of the medial plateau. This asymmetric rolling motion indicated an element of internal tibial rotation. Furthermore, the tibia was found to articulate with the femur at the most proximal points of the condyles in deep flexion. These data on the kinematics and contact characteristics of the tibio-femoral joint must be considered in any approach to design for a Deep Flexion Knee Implant. PMID- 9728682 TI - A new method to measure elastic properties of plastic-viscoelastic connective tissue. AB - An experimental protocol was tested to measure elastic properties of connective tissue displaying viscoelastic as well as plastic properties. The protocol consisted of a slow rate, linear elongation (0.88 mms(-1), 8 mm) in combination with a superimposed sinusoidal vibration of small amplitude (50 Hz, 0.1 mm). Using digital filters and mathematical algorithms, the force responses to linear elongation and to vibration were obtained. The method was tested on excised fibromuscular tissue of the vaginal wall obtained from women who suffered a vaginal prolapse. The force-stiffness and force-elongation relationships based on the vibration response were unaffected by any long-term deformation of the specimens. The directly measured force-elongation curves were strongly affected by these deformations. It was therefore concluded that with the new method, it is possible to determine the elastic properties accurately. Furthermore, this method seems more sensitive to small changes in elastic properties than the classic tensile test. PMID- 9728683 TI - Estimation of parameters in a two-pool urea kinetic model for hemodialysis. PMID- 9728684 TI - Isolation and partial characterization of a thermostable extracellular protease of Bacillus polymyxa B-17. AB - Bacillus polymyxa B-17, a sporeforming psychrotroph produced a thermostable protease. The protease was purified to homogeneity from cell free broth culture by precipitation with ammonium sulfate and gel filtration through Sephadex G-100. The enzyme had a temperature optimum at 50 degrees C and shared significant activity at 70 degrees C. The protease was also active over a wide range of pH, 5.5 to 10.0, and had optimum activity at pH 7.5. It was inhibited by metal chelating agents and has a molecular weight of 30 kDa. PMID- 9728685 TI - Characterization of a bacteriocin produced by Streptococcus thermophilus 81. AB - A new bacteriocin, produced by Streptococcus thermophilus 81 has been isolated, purified and characterized. By its heat sensitivity and broad inhibitory spectrum it does not resemble any other S. thermophilus bacteriocin. The mode of action is bacteriostatic. This peptide of 32 amino acids is efficient against several Bacillus species, Listeria monocytogenes, Salmonella typhimurium, Escherichia coli, Yersinia pseudotuberculosis and Yersinia enterocolitica. This bacteriocin is heat labile but its activity was not altered by pH variation from 3 to 10. Six months of storage at 40 degrees C did not influence the activity. The inactivation by detergents and the inability to resolve the protein in SDS-PAGE supposes a more complex structure or a possible stabilizing effect of other molecules. The low sensitivity of Lactobacillus delbrueckii subsp. bulgaricus to the isolated bacteriocin suggests that S. thermophilus 81 may be used in yoghurt starters. PMID- 9728686 TI - Degradation of pectic compounds during pasteurised vegetable juice spoilage by Chryseomonas luteola: a predictive microbiology approach. AB - Predictive modelling consists in describing effects of environmental factors on microbial growth parameters. With food spoilage bacteria, this approach must be extended to both growth and food damage characterisation. In order to study the incidence of storage temperature on vegetable damage, using predictive microbiology tools, kinetics of pectic compound degradation were studied. Chryseomonas luteola has been chosen because of its ability to grow on post harvested vegetables. Experiments were performed at refrigerated temperatures (0 10 degrees C) with low initial bacterial charges (10(1)-10(3.5) cfu/ml). Microbial specific growth rate (mu), stability phase before pectic degradation (Sp) and alteration percentage (Ap) were chosen as reference parameters. Then, sub-optimal temperature effects on these three parameters were estimated using modified Ratkowsky model. Results obtained in synthetic medium were compared with data observed in endive juice to appreciate the alteration of vegetable during post-harvest storage. PMID- 9728687 TI - Survival of Vibrio spp. including inoculated V. cholerae 0139 during heat treatment of cockles (Anadara granosa). AB - The effect of heat-treatment on the internal temperature of raw cockles (Anadara granosa) and survival of their intrinsic flora of Vibrio spp. as well as of inoculated V. cholerae 0139 was examined. The cockles were purchased from markets in Malaysia and had an average weight including shells of 8.90+/-2.45 g. In one experiment heatpenetration of individual cockles was examined. Cockles weighing < 8 g (including shell) exhibited maximum internal temperatures of between 50 and 75 degrees C when heated in water at 99 degrees C for 10 s and 71-93 degrees C when heated for 30 s. Cockles weighing > 12 g exhibited maximum internal temperatures between 42 and 58 degrees C when heated in water at 99 degrees C for 10 s and 56-69 degrees C when heated for 30 s. In another experiment, heat treatment of 10 cockles treated as a group at 99 degrees C for 10 or 30 s resulted in reduction of levels of intrinsic Vibrio spp. (enumerated directly on thiosulphate-citrate-bile salt sucrose agar; TCBS) from 5.73 to 3.15 log cfu g( 1) or below 1 log cfu g(-1), respectively. The levels of Vibrio spp. after heat treatment decreased with an increase in numbers of cockles grouped together during treatment. In a third experiment V. cholerae 0139 was inoculated into cockles and subjected to heat-treatment at 99 degrees C for 0, 10, 15, 20, 25 or 30 s. The levels of Vibrio spp. in uninoculated, non-heat-treated cockles was 4.89 log cfu g(-1) on TCBS, and the predominant species were V. parahaemolyticus and V. alginolyticus. V. cholerae 0139 inoculated into cockles with an average weight of 13.5+/-1.90 g (including shell) decreased for samples examined immediately after heat-treatment from 6 log cfu g(-1) initially to 3.5 log cfu g( 1) after 25 s and < 1 log cfu g(-1) (TCBS) after 30 s of heat-treatment. The most probable number method by enrichment in alkaline peptone water gave in general within 1 log unit higher counts than TCBS direct enumeration. TCBS direct enumeration and MPN counts were up to 2.38 or 1.30 log units higher, respectively, for samples heat-treated for 20 s or longer and stored for 6 h at 30 degrees C before examination, than for samples heat-treated for same periods of time and examined immediately. This study shows that a mild heat-treatment of cockles for up to 25 s is inadequate to ensure a large reduction in numbers of Vibrio spp., including V. cholerae 0139. PMID- 9728688 TI - Microbial contamination of meat during the skinning of beef carcass hindquarters at three slaughtering plants. AB - The microbiological effects on the product of the series of operations for skinning the hindquarters of beef carcasses at three packing plants were assessed. Samples were obtained at each plant from randomly selected carcasses, by swabbing specified sites related to opening cuts, rump skinning or flank skinning operations, randomly selected sites along the lines of the opening cuts, or randomly selected sites on the skinned hindquarters of carcasses. A set of 25 samples of each type was collected at each plant, with the collection of a single sample from each selected carcass. Aerobic counts, coliforms and Escherichia coli were enumerated in each sample, and a log mean value was estimated for each set of 25 counts on the assumption of a log normal distribution of the counts. The data indicated that the hindquarters skinning operations at plant A were hygienically inferior to those at the other two plants, with mean numbers of coliforms and E. coli being about two orders of magnitude greater, and aerobic counts being an order of magnitude greater on the skinned hindquarters of carcasses from plant A than on those from plants B or C. The data further indicated that the operation for cutting open the skin at plant C was hygienically superior to the equivalent operation at plant B, but that the operations for skinning the rump and flank at plant B were hygienically superior to the equivalent operations at plant C. The findings suggest that objective assessment of the microbiological effects on carcasses of beef carcass dressing processes will be required to ensure that Hazard Analysis: Critical Control Point and Quality Management Systems are operated to control the microbiological condition of carcasses. PMID- 9728689 TI - Effect of water activity on hydrolytic enzyme production by Fusarium moniliforme and Fusarium proliferatum during colonisation of maize. AB - The effect of different water availabilities (water activity, aw; 0.98-0.93) and time (up to 15 days) on the production of seven hydrolytic enzymes by strains of F. moniliforme and F. proliferatum during early colonisation of gamma-irradiated living maize grain were examined in this study. Both the total activity (micromol 4-nitrophenol min(-1) g(-1) maize) and specific activity (nmol 4-nitrophenol min( 1) microg(-1) protein) were quantified using chromogenic p-nitrophenyl substrates. The dominant three enzymes produced by the fungi on whole colonised maize kernels were alpha-D-galactosidase, beta-D-glucosidase, and N-acetyl-beta-D glucosaminidase. The other four enzymes were all produced in much lower total amounts and in terms of specific activity (beta-D-fucosidase, alpha-D mannosidase, beta-D-xylosidase and N-acetyl-alpha-D-glucosaminidase), similar to that in uncolonised control maize grain. There were significant increases in the total production of the three predominant enzymes between 3-15 days colonisation, and between 3-6 days in terms of specific activity when compared to untreated controls. The total and specific activity of the alpha-D-galactosidase, beta-D glucosidase and N-acetyl-beta-D-glucosaminidase, were maximum at 0.98 aw with significantly less being produced at 0.95 and 0.93 aw, with the exception of the total activity of alpha-D-galactosidase which was similar at both 0.95 and 0.93 aw. Single factors (time, aw, and inoculation treatment), two- and three- way interactions were all statistically significant for the three dominant enzymes produced except for specific activity of beta-D-glucosidase (two and three-way interactions) and for total activity of alpha-D-galactosidase in the time x aw treatment. This study suggests that these hydrolytic enzymes may play an important role in enabling these important fumonisin-producing Fusarium spp. to rapidly infect living maize grain over a wide aw range. PMID- 9728690 TI - Amount of enterotoxigenic Clostridium perfringens in meat detected by nested PCR. AB - The incidence and quantity of enterotoxigenic Clostridium perfringens in beef, pork, and chicken meat were determined and compared with that of the total enterotoxigenic and nonenterotoxigenic C. perfringens. The method for the detection and quantification of enterotoxigenic C. perfringens consisted of a combination of the most probable number (MPN) method and a nested polymerase chain reaction after culturing of the sample. The results obtained by this method for inoculated meat samples were significantly correlated with those obtained by the plate count method. When the method was applied to the detection and quantification of enterotoxigenic C. perfringens found in randomly selected meat samples, the organism was found in 2% of the beef pieces (< 10(2) MPN/100 g) and 12% of the chicken pieces (< 10(2)-4.3 x 10(2) MPN/100 g) out of the 50 pieces of each meat tested. No enterotoxigenic C. perfringens was found in pork. Total C. perfringens was found in 16% of the beef (< 10(2)-4.3 x 10(2) MPN/100 g), 10% of the pork (< 10(2) MPN/100 g), and 84% of the chicken (< 10(2)-9.3 x 10(3) MPN/100 g) when 50 pieces of each meat was tested by the conventional MPN method. As shown in the above methods, the majority of cells were not enterotoxigenic cells in the population of C. perfringens. A small number of enterotoxigenic cells of C. perfringens co-existed with a large number of nonenterotoxigenic cells in the same meat sample. PMID- 9728691 TI - A comparison of Listeria monocytogenes serovar 4b isolates of clinical and food origin in Japan by pulsed-field gel electrophoresis. AB - Pulsed-field gel electrophoresis (PFGE) patterns of 102 L. monocytogenes serovar 4b isolates from patients and foods examined in Japan were compared with 16 isolates from foodborne listeriosis episodes which occurred in North America or Europe. Using a combination of PFGE patterns with the restriction enzymes SmaI, ApaI, AscI and Sse8387I, 82 clinical isolates from Japan were categorized into 45 PFGE types: the largest group of 17 isolates (20.7%) were of the same PFGE type as cultures from the large foodborne outbreaks which occurred in California (1985) and Switzerland (1983-1987). Twenty cultures from foods on retail sale in Japan were classified into 12 PFGE types: four isolates were of three PFGE types also recognized among isolates of clinical origin from Japan, including the predominant clinical type. PMID- 9728692 TI - A rapid method for the identification and partial serotyping of Listeria monocytogenes in food by PCR and restriction enzyme analysis. AB - Two highly specific primers for Listeria monocytogenes were used to yield from foods such as milk, soft cheese and meat, PCR products that were cleaved with the restriction enzyme HindIII. The fragments generated allowed a distinction between two groups of L. monocytogenes serovars: serovars 1/2a and 1/2c cluster in one group and serovars 1/2b, 3b and 4b in the other subgroup. Since this procedure can be completed in 24 h, an epidemiological association between human disease and suspected sources can be rapidly confirmed at the subgroup level in the laboratory. PMID- 9728693 TI - Effect of nitrate and nitrite curing salts on microbial changes and sensory quality of non-fermented sausages. AB - The effects of nitrate and nitrite curing salts on microbial changes and sensory quality of non-fermented sausages of small diameter were investigated. During pre ripening (day 5), levels of lactic acid bacteria and yeasts were slightly higher in nitrite-made sausages than in those made with nitrate. In contrast, nitrite discouraged the growth of psychrotrophs as occurs in fermented sausages. By the end of ripening (day 26), levels of microorganisms were similar in both batches of sausages except for psychrotrophs being higher in those made with nitrite. Nitrate-made sausages showed higher aroma and taste intensity. PMID- 9728695 TI - Modification of Saccharomyces cerevisiae thermotolerance following rapid exposure to acid stress. AB - Thermotolerance was induced in cells of Saccharomyces cerevisiae YPH499 pre exposed, during 10 min and in the presence of glucose, to a mild acid-stress with HCl at pH 3.5. Thermotolerance was not induced in cells exposed to a severe acid stress by 50 mM acetic acid at pH 3.5, or HCl at pH 2.5 or pH 2.0. Yeast cells pre-incubated under glucose starvation were found to be more tolerant to a lethal heat stress than cells pre-incubated in glucose-supplemented media, despite the pH value of the media (range 2.0-6.5) and the type of acidulant used (HCl or acetic acid). Moreover, the high thermotolerance exhibited by cells pre-incubated at pH 6.5 for 10 min under glucose starvation was not significantly modified by the acidification of the pre-incubation medium. Results are discussed based on the effect that glucose and a mild or severe acid stress have on plasma membrane H+-ATPase activity and on cytosolic pH values, estimated in a previous work. PMID- 9728694 TI - Influence of water activity on Penicillium citrinum growth and kinetics of citrinin accumulation in wheat. AB - The influence of water activity (aw) on both Penicillium citrinum growth and citrinin accumulation in wheat was studied. Wheat conditioned at different levels of aw and inoculated with a citrinin producer strain was incubated at 30 degrees C for 2 months. Fungal growth was assessed by microscopic examination. P. citrinum grew down to aw 0.775. Citrinin was not detected in the substrate at aw 0.800 and lower. As aw increased the toxin was detected earlier and the maximum accumulation increased markedly (65 microg/kg at aw 0.810, 460 microg/kg at aw 0.825 and 22 mg/kg at aw 0.885). Citrinin concentration declines rapidly after reaching the maximum at each aw level. PMID- 9728696 TI - Midfoot fusion technique for neuroarthropathic feet: biomechanical analysis and rationale. AB - To test the hypothesis that a plate applied to the plantar (tension) side of the medial midfoot provides stronger fixation than midfoot fusion with screw fixation, we biomechanically compared the two constructs for midfoot fusion. We created a model of midfoot instability in eight matched pairs of cadaver legs by section of joint capsule, ligaments, and tendons about Lisfranc's joints, and then performed a load-to-failure study to compare the fixation provided by a plantarly applied third tubular plate with that by cortical screws. After an initial load deformation curve to 1000 N was obtained, specimens were cyclically loaded at 200 to 750 N for 3000 cycles and then loaded to failure (screw pullout, fracture, or deformation >3 mm). Comparing the plantar plate and midfoot fusion with screw fixation constructs, a plate applied to the plantar (tension) aspect of the medial midfoot provides a stronger, sturdier construct than does midfoot fusion with screw fixation. PMID- 9728698 TI - Pathology of the posterior tibial tendon in posterior tibial tendon insufficiency. AB - Gross and histologic examinations of 15 normal cadaver and 15 surgical posterior tibial tendons from patients with posterior tibial tendon insufficiency were performed. All surgical specimens were abnormal with enlargement distal to the medial malleolus and a dull white appearance. At histologic examination, 12 of 15 cadaver tendons displayed normal tendon structure characterized by linear orientation of collagen bundles, normal fibroblast cellularity, low vascular density, and insertional chondroid metaplasia. The surgical specimens displayed a degenerative tendinosis characterized by increased mucin content, fibroblast hypercellularity, chondroid metaplasia, and neovascularization. This resulted in marked disruption of the linear orientation of the collagen bundles. PMID- 9728697 TI - Isolated subtalar arthrodesis. AB - Forty-eight isolated subtalar arthrodeses in 44 patients with an average follow up of 59.5 months were retrospectively reviewed. Original diagnoses included talocalcaneal coalition, healed calcaneal fracture with subtalar arthrosis, acquired flatfoot because of posterior tibial tendon dysfunction, degenerative subtalar arthrosis, subtalar instability, and psoriatic arthritis. Ninety-three percent of patients were very satisfied or satisfied with their treatment. Pain and function improved significantly, and the American Orthopaedic Foot and Ankle Society ankle-hindfoot score at follow-up was 89. There were six unsatisfactory results: three feet had calcaneal fractures and three were malpositioned. Union was achieved in all cases. Transverse tarsal motion was diminished by 40%, dorsiflexion by 30%, and plantarflexion by 9%. There was a 36% and 41% incidence of mild radiographic progression of arthrosis in the ankle and transverse tarsal joint, respectively. Isolated subtalar arthrodesis provided a highly successful result in the disease presented, and this study provides support for the use of a selected hindfoot fusion procedure for specific indications. PMID- 9728699 TI - Histopathological study of nonosseous tarsal coalition. AB - Histopathological analysis was performed on 55 feet in 48 patients with nonosseous tarsal coalitions. Histological findings were similar to those observed at the tendinous attachment site of Osgood-Schlatter disease, accessory navicular, and bipartite patellae. No nerve elements were observed in the fibrocartilaginous tissue at the coalition. Nerve elements were present only in periosteum and articular capsule surrounding the coalition. Pain in the tarsal coalition is not mediated by nerve elements at the coalition site itself. It is assumed that the pain is caused by mechanical abnormality that results from incomplete coalition. Incomplete coalition produces microfractures and remodelings on the boundaries between bone and the coalition, which then lead to degenerative changes. This mechanical abnormality seems to induce pain via free nerve endings in the periosteum and in the articular capsule surrounding the coalition. PMID- 9728700 TI - Metatarsophalangeal and intermetatarsal angle: different values and interpretation of postoperative results dependent on the technique of measurement. AB - For measurement of the first metatarsophalangeal angle and intermetatarsal angle I-II, five different methods for drawing the axis of the first metatarsal have been published. This study aimed to evaluate differences in the resulting angles that depend on the method of drawing this axis. Using pre- and postoperative radiographs of 20 patients who had surgery on the hallux (chevron procedure), highly significant differences were found: mean values for the preoperative metatarsophalangeal angle ranged from 27.3 degrees to 31.9 degrees; the mean postoperative values were calculated at 8.6 degrees to 20.3 degrees. The preoperative mean of intermetatarsal angle I-II showed values from 13.0 degrees to 17.6 degrees; the postoperative mean ranged from 5.2 degrees to 16.7 degrees. These differences--especially in the postoperative evaluation--resulted in a postoperative improvement between 11.6 degrees and 20.8 degrees for the metatarsophalangeal angle and between 0.9 degrees and 10.0 degrees for the intermetatarsal angle. These wide differences seem to be unacceptable for angles as a criterion of success in surgery on the hallux. The reason for these discrepancies can be found in the different relations of the points of reference to the anatomical outline of the metatarsal and the site of osteotomy. As a consequence of this study, defining the axis of the first metatarsal as a line connecting the center of the articular surface of the metatarsal head and the center of the proximal articulation can be recommended as the most appropriate method. The resulting angles are independent of the type of surgery performed on the hallux. PMID- 9728701 TI - Abduction stress and AP weightbearing radiography of purely ligamentous injury in the tarsometatarsal joint. AB - Twenty volunteers (40 feet) with no prior foot injury underwent standardized abduction stress and standing AP radiographs. Subsequently, the Lisfranc and dorsal tarsometatarsal ligaments in nine feet from cadavers were sectioned in a varying sequential manner, and interval standardized radiographs of abduction stress and AP simulated weightbearing were obtained. On abduction stress radiographs in 39 of 40 feet of volunteers and nine of nine feet of cadavers before sectioning, a line tangential to the medial aspect of the navicular and medial cuneiform (medial column line) intersected the base of the first metatarsal. Combining the sectioning of the Lisfranc and dorsal tarsometatarsal ligaments produced a disruption of the medial column line in all feet from cadavers. Disruption of this medial column line may be a simple and valuable diagnostic tool for determining significant ligamentous injury to the tarsometatarsal interval. PMID- 9728702 TI - Measurement of dorsal mobility in the first ray: elimination of fat pad compression as a variable. AB - Previous designs for a device to measure first ray mobility have included compression of the first metatarsal fat pad as part of the measurement of displacement or have failed to standardize the force applied to the head of the first metatarsal. In this investigation, assessment of vertical mobility of the first ray of both feet in 14 volunteers was determined using a device that applied dorsiflexing force to the first metatarsal. First ray displacement was measured initially from the plantar surface and then from the dorsal aspect of the head of the first metatarsal. The difference between plantar- and dorsal surface-measured vertical displacement was highly significant. This study suggests that mobility of the first ray measured from the dorsal aspect of the first metatarsal head eliminated compression of the plantar fat pad from being interpreted as part of the measurement of displacement. PMID- 9728703 TI - Assessing sesamoid subluxation: how good is the AP radiograph? AB - Subluxation of the metatarsosesamoid joints frequently occurs with the development of hallux valgus deformity, and the restoration of a normal metatarsosesamoid articulation has been proposed as essential for achieving a biomechanically sound operative result. The position of the sesamoid bones on the AP radiograph is used often to assess the pre- and postoperative relationship between the hallucal sesamoids and the metatarsal sulci. We evaluated the validity of this approach. Thirty subjects with hallux valgus and 30 control subjects participated in this study by undergoing both AP and tangential weightbearing radiographs. The sesamoid station on the AP radiographs was compared with the position of the sesamoids on tangential radiographs, using a new continuous measure to estimate subluxation. In approximately half of the cases, we found a difference between the apparent sesamoid station on the AP radiograph and the true position on the tangential one. Increased metatarsal rotation was associated with misclassification of the sesamoid station on the AP radiograph. We conclude that the standard method for measuring the sesamoid station on the AP radiograph is not valid. Surgeons wishing to evaluate the metatarsosesamoid joint should obtain weightbearing tangential radiographs. PMID- 9728705 TI - The effect of musculus extensor digitorum brevis transfer for chronic lateral ankle instability. AB - Thirty-eight patients who had undergone surgery for instability of their ankles between 1980 and 1994 answered questionnaires regarding the results. In 32 of the 38 patients, clinical examinations were performed including practice on a balancing board, circle-running tests, and active and passive electromyographic measurements on the musculus extensor digitorum brevis (MEDB). The questionnaires showed that 90% of the patients with a median observation time of 9 years (range, 16 months-14 years) were content with the results of their operations, and the number of painful distortions of the ankle were reduced considerably in 96% of the patients. Within the group of active sportspersons with more than 4 hours of weekly exercise, 42% returned to their previous levels of sports activity. The tendency to have ankle pain disappeared in 90% of the patients. At the clinical examination, the time spent on the balancing board was reduced by 25% for the operated foot. When the patient ran in a circle with the operated leg toward the center, as compared with running with the nonoperated leg toward the center, the time was enhanced by 8%. We found electromyographic activity in the MEDB during active movement of the toes and with passive supination of the talocrural joint but not during passive pronation of the foot. The MEDB transfer procedure not only strengthens the lateral ligaments but also seems to add proprioceptive protection to the ankle to prevent distortions. PMID- 9728704 TI - Ankle fracture classification: a comparison of reliability of three X-ray views versus two. AB - Our hypothesis was that malleolar ankle fractures could be classified with two radiographic views as reliably as with three views. Four different observers independently evaluated 99 sets of ankle radiographs. The examiners classified the ankle fractures by using both the Lauge-Hansen and Danis-Weber systems. The interobserver and intraobserver variations were analyzed by kappa statistics. With regard to intraexaminer reliability, the examiners demonstrated excellent accord in classifying the fractures in the Danis-Weber system with either three views or two views. The kappa values were comparable. In the Lauge-Hansen system, three examiners demonstrated excellent accord and one examiner demonstrated good accord in classifying the fractures. Similar kappa values were generated when examiners classified fractures with either three views or two views. With regard to interexaminer reliability, good to excellent accord was demonstrated overall among the four examiners when they used the Danis-Weber system with either three views or two views. The examiners were in good agreement when they used the Lauge Hansen system. Similar kappa values were generated whether the examiners used three views or two views. Three radiographic views are usually ordered for evaluation of an acute ankle injury. Previous studies have shown that only two views are needed for diagnosis of a malleolar ankle fracture. This study demonstrates that malleolar ankle fractures can be classified with two views, lateral or mortise, with a reliability as good as that achieved with three views. The best agreement is achieved with lateral and mortise views. PMID- 9728706 TI - Hypertrophic extensor digitorum brevis muscles simulating pseudotumors: a case report. AB - Pseudotumors can involve soft tissue or bone, and can lead to difficulty or uncertainty in diagnosis and treatment. The authors describe bilateral hypertrophy of the belly of the extensor digitorum that simulated ganglia. PMID- 9728708 TI - Supination lag as an indication of posterior tibial tendon dysfunction. PMID- 9728707 TI - Closed reduction of a lesser toe fracture. PMID- 9728709 TI - Clinical application of shape memory staples. PMID- 9728710 TI - An evaluation of the use of gait plate inlays in the short-term management of the intoeing child. PMID- 9728712 TI - Orthopedists are physicians, not surgical technicians. PMID- 9728711 TI - Effect of medial displacement calcaneal osteotomy on ankle kinematics in a cadaver model. PMID- 9728713 TI - Hemarthrosis and tourniquet use. PMID- 9728714 TI - A comparison of MRI findings in patients with acute and chronic ACL tears. AB - This retrospective study compared the magnetic resonance imaging (MRI) findings in 87 patients with acute and chronic anterior cruciate ligament (ACL) tears. Sixty patients had acute tears and 27 had chronic tears. The appearance of the torn ligament was examined on MRI, and associated meniscal and osteochondral injuries were described. All findings were verified at arthroscopy. Acute ACL tears (MRI examination was performed within 6 weeks of injury) were typified by the presence of diffuse (58%) or focal (42%) increased signal within the ligament, whereas chronic ACL tears (MRI examination was performed more than 6 months after injury) usually appeared as either a fragmented ligament (44%) or an intact band of low signal with abnormal orientation (30%). Patients with chronic ACL tears had a higher prevalence of medial meniscal tears (78% versus 40%), articular chondromalacia, and an increased posterior cruciate bow ratio (0.47 versus 0.37) in association with chronic ACL tears. A bone bruise was seen in 68% of acute ACL tears but in no case of chronic ACL tear. On MRI, there are salient differences between acute and chronic ACL tears. Chronic ACL tears are associated with a greater prevalence of meniscal and osteochondral injuries. These findings may have implications for future treatment recommendations. PMID- 9728715 TI - Anatomically based patellar resection criteria for total knee arthroplasty. AB - Thirty-six patellae were used to geometrically assess anatomically based patellar resection criteria for total knee arthroplasty. The plane of resection was defined in the coronal plane by three distinct anatomic landmarks on the undersurface of the patella: the medial and lateral edges of the quadriceps tendon insertion and the lateral edge of the patellar ligament insertion. The depth of the resection plane is defined by the anatomic landmark points approximately 1 mm posterior to the posterior aspect of the superior and inferior ligament insertions. Following resection, the resected patellae were geometrically characterized using two dimensional rectangular Cartesian coordinate system. The anatomically based patellar resection yielded a simple, consistent, and reliable method for patellar resection for total knee arthroplasty. The proportionate geometric characteristics of both the remaining anterior piece and the resected posterior piece were remarkably uniform. Following resection, the remaining patellar thickness was 65.6+/-5.3% (mean+/-SD) of the original thickness. The maximum width to height ratio of the resected surface was approximately 1:1. The thickness to diameter ratio of the resected portion of the patella was approximately 1:5. The apex of the patella was slightly off to the medial-distal quadrant from the geometric center of the resected surface. The results of this study indicate that anatomically based patellar resection criteria may help reduce the surgical variations associated with patellar resurfacing in total knee arthroplasties with dome-shaped prostheses. PMID- 9728716 TI - Effect of Elmslie-Trillat and Roux-Goldthwait procedures on patellofemoral relationships and symptoms in patients with patellar dislocations. AB - Seventeen patients (18 knees) with a history of one or more patellar dislocations underwent Elmslie-Trillat (9 knees [group 1]) or Roux-Goldthwait (9 knees [group 2]) patellar realignment surgery. Anterior knee pain was evaluated with a questionnaire, and lateral patellar displacement and tilt were analyzed by magnetic resonance imaging both pre- and postoperatively. Postoperative evaluations were performed after 44 and 50 months (mean) in groups 1 and 2, respectively. The Elmslie-Trillat procedure provided better subjective relief of anterior knee pain and symptoms. One redislocation occurred in each group. Both procedures relieved excessive patellar lateralization, but the effect on patellar tilt was less marked. In patients who underwent the Roux-Goldthwait operation, a less pronounced correction of tilt and lateralization appeared to correlate with more satisfactory subjective results. PMID- 9728717 TI - Selective denervation of the knee: experience, case reports, and technical notes. AB - This article outlines the indications and technique of selective denervation for chronic knee pain secondary to neuromata from prior surgery or trauma and describes the results obtained in a series of patients who underwent selective denervation for neuromatous knee. Of the 13 patients in this series, 3 (23%) rated their outcome as excellent, 7 (54%) rated the outcome as good, and 3 (23%) rated the outcome as poor; no patient rated the outcome as worse. Several case reports are included to illustrate the procedure. This technique should be considered an option in select patients with neuromatous knee pain. PMID- 9728718 TI - The skeletally fixed knee hinge for the grossly unstable knee. AB - This study examined whether a skeletally fixed prefabricated knee hinge can provide the intact or unstable knee with normal motion through a specific arc of motion. Eight cadaveric knee specimens were used. The amount of motion mismatch between knee and hinge motion was evaluated at six different knee flexion angles. With all knee ligaments intact, addition of the hinge resulted in increasing amounts of joint compression with knee flexion. When all knee ligaments were cut, there was some degree of distraction with 0 degrees of knee flexion, which seemed to gradually decrease and become compressive at 80 degrees of flexion. These values were not statistically significant. In contrast, the mismatch between anterior and posterior tibial translation mismatch was statistically significant. With the ligaments intact, the addition of the hinge resulted in increased amounts of posterior tibial translation, which became significant at 80 degrees of flexion. Similarly, when the ligaments were cut with the hinge intact, there was an increasing amount of posterior tibial translation, which became significant at 60 degrees of flexion. There was also a significant amount of anterior tibial translation at 0 degrees in this group. These results indicate that the hinge allows only a limited range of motion that does not significantly alter tibial translation or joint compression or distraction. Whether this amount of motion is enough to improve the outcome of the grossly unstable knee is unknown. The use of a more sophisticated hinge system might accomplish a greater range of anatomic motion before significant mismatch occurs between hinge and knee motion. PMID- 9728719 TI - Bilateral bony avulsion at the inferior patellar pole in a patient with jumper's knee. PMID- 9728720 TI - Traumatic proximal tibial fracture following anterior cruciate ligament reconstruction. PMID- 9728721 TI - The National Acoustic Laboratories' procedure for selecting the saturation sound pressure level of hearing aids: theoretical derivation. AB - This paper presents the derivation of a procedure for prescribing the saturation sound pressure level (SSPL) of hearing aids. The procedure is designed to be used with either measured values of loudness discomfort level (LDL) or with hearing threshold values alone. SSPL needs to be low enough to prevent the hearing aid from causing loudness discomfort to the aid wearer but high enough to prevent the hearing aid from being excessively saturated by speech. The maximum SSPL likely to be acceptable can be predicted by measuring LDL or by estimating LDL from hearing thresholds. The minimum SSPL likely to be acceptable can be predicted by calculating, for any particular hearing loss, the amount of gain likely to be needed and hence the SSPL needed if the speech input signal is continuous discourse at an overall level of 75 dB SPL. The midpoint between the minimum and maximum acceptable SSPL values is defined as the optimal or prescribed SSPL, and the three frequency average (3FA; 500, 1000, and 2000 Hz) value of this can be predicted from the 3FA hearing thresholds. Alternatively, the SSPL prescription at each frequency can be prescribed on the basis of the hearing aid gain at each frequency. For either method, the SSPL prescription needs to be increased for people with a conductive component to their hearing losses. The SSPL prescription, when referred to a 2-cc coupler, needs to be decreased for infants, for deeply inserted hearing aids, for multichannel hearing aids that limit SSPL separately in each band, and possibly for nonlinear hearing aids. The 3FA SSPL prescribed for persons with a sensorineural hearing loss increases linearly from 89 dB SPL for normal hearing to 107 dB SPL for a person with a 60 dB HL 3FA loss, and then linearly again to 139 dB SPL for a person with a 120 dB HL 3FA loss. The procedure predicts that the acceptable range of SSPLs is very wide for people with mild losses but is vanishingly small for people with profound losses. PMID- 9728722 TI - The National Acoustic Laboratories' procedure for selecting the saturation sound pressure level of hearing aids: experimental validation. AB - OBJECTIVE: The primary aim of this study is to evaluate the accuracy of a new procedure for selecting the saturation sound pressure level (SSPL) of hearing aids. Secondary aims are to investigate what limits the minimum SSPL that is acceptable to clients and whether the type of limiting (peak clipping or compression limiting) affects the SSPL required. DESIGN: The study comprised two experiments. In the first, subjects increased the SSPL of a laboratory master hearing aid until they experienced loudness discomfort and decreased it until the sound became less acceptable in some way. In the second study, subjects wore multi-memory programmable hearing aids in their own environments and reported which of the two programs, differing only in SSPL setting, provided the more acceptable sound quality and comfort. RESULTS: The theoretical procedure being investigated prescribed SSPLs that were within the acceptable range for 86% of the subjects in the laboratory study and for 63% of the subjects in the field experiment. On average, the theoretical predictions were neither too high nor too low. Incorporating individual measurements of loudness discomfort level into the prescription formula increased accuracy by such a small amount that it was not considered worthwhile. For a compression limiting hearing aid, the first thing that subjects noticed as SSPL was reduced was inadequate loudness. For the peak clipping hearing aid, however, both inadequate loudness and perception of distortion limited the acceptable SSPL range. CONCLUSION: The theoretical procedure provides a good initial prescription of three frequency average SSPL, but it is still essential to evaluate the fitting and, if necessary, fine tune the individual's hearing aid. Compression limiting hearing aids can have slightly lower SSPL settings than peak clipping hearing aids for the same acceptability. PMID- 9728723 TI - Comparison of two digital hearing aids. AB - OBJECTIVE: The objective of this investigation was to compare real and perceived benefit for two currently marketed digital hearing aids, the Oticon DigiFocus and the Widex Senso. The hearing aids have different philosophies of design and fitting strategies; as a result, it was hypothesized that there would be performance differences. DESIGN: Twenty subjects with documented sensorineural hearing losses were fit with each of the two digital hearing aids. After 4 wk of use with each hearing aid, a battery of objective and subjective tests was completed to assess hearing aid benefit. RESULTS: No significant differences were found between the hearing aids as revealed by the objective testing of speech recognition and self-report inventories of hearing aid benefit. The DigiFocus was shown by real ear measurements to provide more high-frequency gain than the Senso. The Widex Senso was preferred by 13 of the 20 subjects (seven of 10 of the new hearing aid users). This may be explained, in part, by the increased high frequency gain provided by the Oticon DigiFocus, which was perceived as having greater "harshness." CONCLUSIONS: Based on the results of this investigation, neither hearing aid processor was shown to be superior to the other. In addition, the least amount of objective benefit was shown in the presence of background noise. PMID- 9728724 TI - Cortical evoked response to acoustic change within a syllable. AB - OBJECTIVE: To investigate whether the evoked potential to a complex naturally produced speech syllable could be decomposed to reflect the contributions of the acoustic events contained in the constituent phonemes. DESIGN: Auditory cortical evoked potentials N1 and P2 were obtained in eight adults with normal hearing. Three naturally produced speech stimuli were used: 1) the syllable [sei]; 2) the sibilant [s], extracted from the syllable; 3) the vowel [ei] extracted from the syllable. The isolated sibilant and vowel preserved the same time relationships to the sampling window as they did in the complete syllable. Evoked potentials were collected at Fz, Cz, Pz, A1, and A2, referenced to the nose. RESULTS: In the group mean waveforms, clear responses were observed to both the sibilant and the isolated vowel. Although the response to the [s] was weaker than that to [ei], both had N1 and P2 components with latencies, in relation to sound onset, appropriate to cortical onset potentials. The vowel onset response was preserved in the response to the complete syllable, though with reduced amplitude. This pattern was observable in six of the eight waveforms from individual subjects. CONCLUSIONS: It seems likely that the response to [ei] within the complete syllable reflects changes of cortical activation caused by amplitude or spectral change at the transition from consonant to vowel. The change from aperiodic to periodic stimulation may also produce changes in cortical activation that contribute to the observed response. Whatever the mechanism, the important conclusion is that the auditory cortical evoked potential to complex, time varying speech waveforms can reflect features of the underlying acoustic patterns. Such potentials may have value in the evaluation of speech perception capacity in young hearing-impaired children. PMID- 9728725 TI - Spatial perception of speech in various signal to noise ratios. AB - OBJECTIVE: The study was designed to assess the effects of background noise level on the detection and localization of speech. DESIGN: The phrase "Where is this?" was presented either in quiet or in a diffuse noise field, through loudspeakers arranged in a 360 degrees azimuth array. The noise conditions included 11 signal to noise ratios (SNRs) ranging from -18 dB SNR to +12 dB SNR in 3 dB increments. Seventeen normal-hearing subjects, aged 18 to 29, participated in the study. RESULTS: Results revealed that in all listening conditions the signal was most easily detected when presented through a loudspeaker positioned at 90 degrees or 270 degrees azimuth. Although the actual level for 50% detection varied as a function of loudspeaker location and SNR, 85% and 100% of all presentations of the signal were detected at -9 dB and -6 dB SNR, respectively. Localization accuracy improved as the SNR increased, ranging from 18% accuracy at -18 dB SNR to 89% at +12 dB SNR. Localization accuracy in quiet was 95%. The data are discussed in reference to patterns of responses at each loudspeaker location. CONCLUSIONS: Detection of the target signal deteriorated as background noise level increased and was dependent on the source location of the incoming signal, as expected. Localization accuracy of the target signal was highly dependent on the SNR and spatial location of the signal source. Detection and localization accuracy data were found to be repeatable across test sessions and response patterns were found to be symmetrical on the right and left sides of the horizontal plane. PMID- 9728727 TI - Identification of noise sources that influence distortion product otoacoustic emission measurements in human neonates. AB - OBJECTIVE: The objective of this study was to identify individual sources of noise and their contribution to the overall noise that influences valid measurement of otoacoustic emissions in neonates. The hypothesis was that careful selection of eliciting signals and signal processing parameters, unique analysis of measured results, and control of certain subject characteristics would allow isolation of these individual noise sources and determine their relative influence. DESIGN: Eliciting signal parameters were optimized and held constant to minimize equipment noise. Analysis of noise floors in relation to signal level was used to identify equipment-related noise associated with changes in signal parameters. Analysis of noise floor distributions was used to determine whether environmental noise entered the measurements via inadequate coupling of the probe to the ear. The acoustic characteristics of the middle ear were varied via subject selection to determine the influence of middle-ear characteristics on noise floor levels. RESULTS: The two sources of noise associated with the measurement equipment need not contribute to the noise floor for biologically relevant otoacoustic emissions measurements (eliciting signal levels between 30 and 75 dB SPL). Of the two pathways identified for environmental noise, the pathway resulting from an inadequate seal between the probe and the ear canal can be eliminated. One of the two sources of noise related to the subject, noise resulting from biologic activity unrelated to the ear can be minimized. However, the remaining factor, the status of the middle ear, has been shown to contribute as much as 6 dB to the overall noise floor. CONCLUSIONS: Careful selection of signal parameters and additional data analyses and procedural variables can isolate or control several sources of noise that influence distortion product otoacoustic emission measurements in neonates. Tight coupling between the probe unit and the external ear canal should be maintained for all measurements. Middle ear abnormalities can increase noise floors up to 6 dB. PMID- 9728726 TI - The production of English inflectional morphology, speech production and listening performance in children with cochlear implants. AB - OBJECTIVE: To compare how children who use either cochlear implants (CIs) or hearing aids (HAs) express English inflectional morphemes during conversation, i.e., with voice, with sign, or with both. A secondary objective was to investigate the relationship between morpheme use in pediatric CI users and their speech perception skills, length of experience with the device, and accuracy of phoneme production. DESIGN: Group 1 consisted of 25 children who used CIs, and Group 2 consisted of 13 children who used HAs. All children were prelingually deafened and all used simultaneous communication. A 12 minute spontaneous conversation was elicited, transcribed and coded. Between group comparisons were performed to evaluate differences in modality and number of morphemes used. Additionally, use of morpheme endings was related to length of CI experience, accuracy of phoneme production, and closed-set speech recognition performance. RESULTS: Children who had CI experience produced significantly more English inflected morphemes than children in the HA group. CI participants also expressed the inflected endings by using voice-only mode 91% of the time, whereas HA participants used voice-only mode 1% of the time. In the CI group, a strong relationship was found between number of morpheme endings used and speech recognition scores, length of CI experience and accuracy of phoneme production. The results of this study indicate that input from the CI facilitates children's ability to perceive and comprehend bound morphemes. PMID- 9728728 TI - Prevalence of unilateral hearing loss in children: the National Health and Nutrition Examination Survey II and the Hispanic Health and Nutrition Examination Survey. AB - We compared population-based prevalence rates of unilateral hearing loss among African-American, Cuban-American, Mexican-American, Puerto Rican, and non Hispanic White children 6 to 19 yr of age. The prevalence (per thousand) of overall hearing loss (average decibel HTL >30) ranged from 6.4 in Mexican Americans to 12.3 in Cuban-Americans. The prevalence of moderate to profound unilateral hearing loss (average decibel HTL >50) ranged from 0.0 in Cuban Americans to 5.2 in Puerto Ricans. No statistically significant age or gender differences were found within any of the ethnic groups. Among these five ethnic groups, it is estimated that approximately 391,000 school-aged children in the United States have unilateral hearing loss. PMID- 9728729 TI - The case described by Alois Alzheimer in 1911. Historical and conceptual perspectives based on the clinical record and neurohistological sections. AB - In 1906, Alzheimer presented the first case of the disease which was later named Alzheimer's disease by Kraeplin. While the publication on this case in 1907 is only a relatively short communication, Alzheimer published a very comprehensive paper in 1911 in which he discussed the concept of the disease in detail. This publication focusses on the report of a second patient suffering from Alzheimer's disease, the case of Johann F. The detection of neurohistopathological sections from this patient found among archives at the Institute of Neuropathology of the University of Munich enabled us to reinvestigate this case using modern methods. Neurohistopathologically, the case of Johann F. is "plaque-only" Alzheimer's disease. There is a controversy in the modern literature as to whether these "plaque-only" cases belong to the modern concept of Alzheimer's disease. A careful analysis of all pros and contras in the literature led to the conclusion that plaque-only cases are also an integrative part of the modern Alzheimer disease concept. PMID- 9728730 TI - Early-onset Alzheimer's disease due to mutations of the presenilin-1 gene on chromosome 14: a 7-year follow-up of a patient with a mutation at codon 139. AB - Mutations in the presenilin-1 gene (PS-1 gene) on chromosome 14 have recently been identified as a cause of familial early-onset Alzheimer's disease (EOAD). To our knowledge, only two German EOAD patients with mutations in the PS-1 gene have been identified thus far. Herein we report the case of a German EOAD patient with a family history of dementia and a missense mutation at codon 139 (M139V) of the PS-1 gene. The patient came to our clinic for the first time when he was 44 years old. During the following 7 years, his Mini-Mental State Examination (MMSE) score dropped from 24 to 0. Myocloni were an early neurological symptom that was already present during the first consultation. We could demonstrate that myoclonic activity was of cortical origin using a back-averaging method. Magnetic resonance imaging (MRI) revealed only slight changes in the early stage of the disease. Follow-up MRI studies showed progression of bitemporal ventricular enlargement and progressive frontal and temporal cortical atrophy. Although the majority of EOAD patients belong to the sporadic (non-genetic) type of AD, early onset dementia, early myocloni and a familial history of AD should direct attention to the possibility of a genetic form of AD. PMID- 9728731 TI - Dimensions of schizophrenic positive symptoms: an exploratory factor analysis investigation. AB - Current psychopathology classifies schizophrenic positive symptoms into four groups: delusions, hallucinations, formal thought disorder, and catatonic symptoms. The present study explores the factor structure of different positive symptoms to refine this classification. The 35 positive symptoms of 429 psychiatric patients, consecutively admitted to any of 95 mental hospitals, with diagnosis of the ICD-10 F20 schizophrenia, were studied. After excluding those items with a base rate of 10% or less, factor analysis yielded six factors. The first factor was loaded by most of Schneider's first-rank symptoms and two specific auditory hallucinations; the second by all the catatonic symptoms and incoherence; the third by bodily delusions/hallucinations; the fourth by delusions of persecution and reference; the fifth by grandiose and religious delusions; and the sixth by visual and miscellaneous hallucinations. The finding that schizophrenic positive symptoms may have more than four dimensions suggests the need for reclassification of schizophrenic symptoms and for reconsideration of evidence-based diagnostic criteria for the disorder. PMID- 9728732 TI - Repetition blindness in schizophrenic patients. AB - Repetition blindness is the failure to report the detection of repeated items in rapid visually presented lists. It can be explained in terms of either a processing limitation or an active inhibitory process. In two studies conducted in either English or German language we set out to induce repetition blindness under various conditions in a total of 47 control subjects and 30 schizophrenic patients. The patients displayed the phenomenon to at least the same degree as normal control subjects. These results render unlikely accounts of repetition blindness which involve processes known to be dysfunctional in schizophrenic patients. Moreover, the study provides an example of how the performance of schizophrenic patients can constrain theories of normal cognition. PMID- 9728733 TI - Risk factors for suicides of inpatients with depressive psychoses. AB - Research on identifying the relevant risk factors for suicides is faced with a multitude of methodological problems. The present study attempts to improve on some of these problems and to isolate those risk factors that are accessible in the early stages of the treatment of inpatients. A total of 3792 inpatients with monopolar or bipolar depression were treated during the period 1981-1992. Suicides (n=33) and controls (n=3759) were compared with respect to 77 sociodemographic and anamnestic variables and 195 standardised items of the admission summary. In addition to an analysis of contingency tables a discriminant analysis was performed. The suicide rate of patients with depressive psychosis was 2.7 times higher than the average rate of 0.324% for the entire clinic. Suicidal tendencies on admission proved to be the best predictor with a frequency of 91% in the suicide group and 40% in the control group, previous attempted suicide being the second best predictor. We conclude that the rate of inpatient suicide may have been underestimated for methodological reasons in the past decades. Many of the risk factors discussed in the literature may be of little predictive value at least in the initial stages of hospital treatment. PMID- 9728734 TI - Rebound insomnia after hypnotic withdrawal in insomniac outpatients. AB - The effect of abrupt medication withdrawal (no-pill discontinuation) was investigated in 1507 insomniacs using the patients' self-ratings on visual analogue scales. Drug discontinuation followed a 28-day treatment period with either 7.5 mg zopiclone, 0.25 mg triazolam, 1.0 mg flunitrazepam, or placebo in a randomized, double-blind, parallel group, multicenter study in private practice. Deterioration below individual pretreatment values (no-pill baseline) in at least one of three subjective parameters of sleep quality (sleep latency, total sleep time, nocturnal awakenings) and three parameters of daytime well-being (morning freshness, daytime tiredness, anxiety) were defined as rebound. The number of patients with rebound (rebound rate) was analyzed for every day of a 2-week posttreatment period. The overall rebound rate was higher in the placebo group (p < or = 0.001) than in each group treated with active drugs. Rebound rates affecting sleep quality were higher for placebo than for zopiclone (p < or = 0.001) and for flunitrazepam (p < or = 0.05). Rebound rates were smaller for zopiclone (p < or = 0.001) and flunitrazepam (p < or = 0.01) than for triazolam. Rebound in at least one item per day appeared in 21.5% (sleep quality) and 25.5% (daytime well-being) of the patients. Rebound decreased with increasing numbers of items of sleep quality or daytime well-being. Patients who did not respond to treatment showed higher rebound rates than those who were treatment responders (p < or = 0.001). Concerning treatment nonresponders, highest rebound was seen in the placebo group, whereas rebound was lowest in placebo responders. These results show that pill discontinuation itself may worsen sleep and daytime well being in the sense of a rebound phenomenon. Furthermore, the number of patients with rebound remained at a high and varying level during the whole posttreatment period. This result indicates that a deterioration of sleep after drug withdrawal is not apparent during a few days but may last for longer periods in some patients and is modified by marked night-to-night variations. PMID- 9728735 TI - Perceived mood and skin body temperature rhythm in depression. AB - Eighteen inpatients affected by recurrent major depression were monitored in their clinical and biological features during the acute episode of illness. Diurnal mood variations rated with Visual Analogue Scale (VAS) and diurnal variations of skin body temperature were measured every 2 h consecutively for 2 days. Circadian rhythmicity of the two parameters was evaluated by cosinor analysis separately for each patient. The inspection of the individual cosine fitting shows that patients with a high circadian rhythmicity in perceived severity of symptomatology tend to show low circadian rhythmicity in skin body temperature, whereas patients with a low VAS oscillation tend to display a higher diurnal variation in skin body temperature. A chi-square test confirmed a statistical significance of the discordance between the two rhythms. We discuss our findings hypothesizing a different degree of entrainment of the disease process to the main circadian pacemaker. PMID- 9728736 TI - Forcing a link? PMID- 9728737 TI - Clustering of childhood acute leukaemia: The EUROCLUS Project. AB - The EUROCLUS Project is a collaborative endeavour in which incidence data for 13 35 1 cases of childhood leukaemia (CL) diagnosed between 1980 and 1989 in 17 countries were referenced to 26 425 small geographic areas and tested for evidence of spatial clustering. A second objective of EUROCLUS was to determine whether clustering of CL was associated with community demographic features and/or proximity to putative environmental hazards. The results show statistically significant evidence of clustering, but the magnitude is small (extra-Poisson variability = 1.65% of Poisson variability). Patterns of clustering are associated with population density and other demographic features which could indicate variations in opportunity for exposure to common infections. There is no consistent evidence that clusters are associated with proximity to nuclear facilities or other putative environmental hazards. PMID- 9728738 TI - Occupational cancer risk in pilots and flight attendants: current epidemiological knowledge. AB - Occupational studies of aircrew in civil or military aviation did not receive much attention until the beginning of this decade. Since 1990, a number of epidemiological studies has been published on the cancer risk among flight personnel. Their results are equivocal: elevated cancer risks have been observed in some studies, but not in others. The exposure situation for pilots and flight attendants is unique with respect to several factors and particularly in that cosmic rays contribute substantially to their cumulative radiation dose. The average annual doses received are relatively low, however, and commonly range between 3 and 6 mSv. Results of epidemiological studies are presented as well as information on planned studies. PMID- 9728739 TI - Consideration on a limit for lifetime occupational radiation exposure. AB - Annual dose limits in occupational radiation exposure are merely a secondary constraint in addition to the primary rule of dose reduction and justification. The limits may, therefore, be reached only in rare, special cases. However, in principle, there might be cases in which the annual limit is continuously exhausted throughout a working life; a high total dose of 0.8 Sv could then be reached. In view of this possibility, there have been considerations of an added restriction by limiting the lifetime occupational dose to 0.4 Sv. The implications of such lifetime doses are considered, and it is shown that an exposure up to the maximum of 0.8 Sv will lead to the need for compensation, if a leukaemia were to occur in the exposed worker. A lifetime dose of 0.4 Sv equally spread over a working life will not lead to the critical value of the probability of causation in excess of 0.5. Nevertheless, it could cause such critical values when it is accumulated during shorter periods. More decisive than the probabilities of causation are, however, the absolute numbers of excess cases of leukaemia due to the occupational exposure. It is seen that less than one excess case would be expected if a group of 100 workers were all exposed to the maximum of 0.8 Sv. Since lifetime doses of 0.8 or 0.4 Sv will be accumulated in very few cases and only with special justification, there appears to be little need to introduce a further limit of lifetime exposure in addition to the current annual limit. PMID- 9728740 TI - Trends in infant leukaemia in West Germany in relation to in utero exposure due to Chernobyl accident. AB - A temporary increase in the incidence of infant leukaemia in Greece was reported by Petridou et al., which was attributed to in utero exposure to ionising radiation resulting from the Chernobyl accident. We performed a similar analysis based on the data of the German Childhood Cancer Registry in order to check whether the observation could be confirmed by means of independent data. Applying the same definitions as Petridou et al., we also observed an increased incidence of infant leukaemia in a cohort of children born after the Chernobyl accident. More detailed analyses, regarding areas with different contamination levels and dose rate gradients over time after the accident, showed, however, no clear trend with regard to exposure. It would therefore appear less likely that the observed effect was caused by exposure to ionising radiation due to the Chernobyl accident. PMID- 9728741 TI - Investigation of lipid peroxidation in liposomes induced by heavy ion irradiation. AB - Lipid peroxidation induced by heavy ion irradiation was investigated in 1,2 dilinoleoyl-sn-glycero-3-phosphocholine (DLPC) liposomes. Lipid peroxidation was induced using accelerated heavy ions that exhibit linear energy transfer (LET) values between 30 and 15000 keV/microm and doses up to 100 kGy. With increasing LET, the formation of lipid peroxidation products such as conjugated dienes, lipid hydroperoxides, and thiobarbituric acid-reactive substances decreased. When comparing differential absorption spectra and membrane fluidity following irradiation with heavy ions and x-rays (3 Gy/min), respectively, it is obvious that there are significant differences between the influences of densely and sparsely ionizing radiation on liposomal membranes. Indications for lipid fragmentation could be detected after heavy ion irradiation. PMID- 9728742 TI - Formation of plasmid DNA strand breaks induced by low-energy ion beam: indication of nuclear stopping effects. AB - Plasmid pGEM 3zf(+) was irradiated by nitrogen ion beam with energies between 20 and 100 keV and the fluence kept as 1x10(12)ions/cm2. The irradiated plasmid was assayed by neutral electrophoresis and quantified by densitometry. The yields of DNA with single-strand and double-strand breaks first increased then decreased with increasing ion energy. There was a maximal yield value in the range of 20 100 keV. The relationship between DNA double-strand breaks (DSB) cross-section and linear energy transfer (LET) also showed a peak-shaped distribution. To understand the physical process during DNA strand breaks, a Monte Carlo calculation code known as TRIM (Transport of Ions in Matter) was used to simulate energy losses due to nuclear stopping and to electronic stopping. It can be assumed that nuclear stopping plays a more important role in DNA strand breaks than electronic stopping in this energy range. The physical mechanisms of DNA strand breaks induced by a low-energy ion beam are also discussed. PMID- 9728743 TI - DNA double-strand breaks in mammalian cells exposed to gamma-rays and very heavy ions. Fragment-size distributions determined by pulsed-field gel electrophoresis. AB - The spatial distribution of DNA double-strand breaks (DSB) was assessed after treatment of mammalian cells (V79) with densely ionizing radiation. Cells were exposed to beams of heavy charged particles (calcium ions: 6.9 MeV/u, 2.1.10(3) keV/microm; uranium ions: 9.0 MeV/u, 1.4.10(4) keV/microm) at the linear accelerator UNILAC of GSI, Darmstadt. DNA was isolated in agarose plugs and subjected to pulsed-field gel electrophoresis under conditions that separated DNA fragments of size 50 kbp to 5 Mbp. The measured fragment distributions were compared to those obtained after gamma-irradiation and were analyzed by means of a convolution and a deconvolution technique. In contrast to the finding for gamma radiation, the distributions produced by heavy ions do not correspond to the random breakage model. Their marked overdispersion and the observed excess of short fragments reflect spatial clustering of DSB that extends over large regions of the DNA, up to several mega base pairs (Mbp). At fluences of 0.75 and 1.5/microm2, calcium ions produce nearly the same shape of fragment spectrum, merely with a difference in the amount of DNA entering the gel; this suggests that the DNA is fragmented by individual calcium ions. At a fluence of 0.8/microm2 uranium ions produce a profile that is shifted to smaller fragment sizes in comparison to the profile obtained at a fluence of 0.4/microm2; this suggests cumulative action of two separate ions in the formation of fragments. These observations are not consistent with the expectation that the uranium ions, with their much larger LET, should be more likely to produce single particle action than the calcium ions. However, a consideration of the greater lateral extension of the tracks of the faster uranium ions explains the observed differences; it suggests that the DNA is closely coiled so that even DNA locations several Mbp apart are usually not separated by less than 0. 1 or 0.2 microm. PMID- 9728744 TI - Calculation of boron neutron capture cell inactivation in vitro based on particle track structure and x-ray sensitivity. AB - The Monte-Carlo technique was used to perform quantitative microdosimetric model calculations of cell survival after boron neutron capture irradiations in vitro. The high energy 7Li and alpha-particles resulting from the neutron capture reaction 10B (n,alpha)7Li are of short range and are highly damaging to cells. The biophysical model of the Monte-Carlo calculations is based on the track structure of these a-particles and 7Li-ions and the x-ray sensitivity of the irradiated cells. The biological effect of these particles can be determined if the lethal effect of local doses deposited in very small fractional volumes of the cell nucleus is known. This lethal effect can be deduced from experimental data of cell survival after x-ray irradiation assuming a Poisson distribution for lethal events. The input data used in a PC-based computer program are the radial dose distribution inside the track of the released particles, cell survival after x-ray irradiation, geometry of the tumor cells, subcellular 10B concentration, and thermal neutron fluence. The basic concept of this Monte-Carlo computer model is demonstrated. Validations of computer calculations are presented by comparing them with experimental data on cell survival. PMID- 9728745 TI - Determination of soil-to-plant transfer factors of 137Cs and 90Sr in the tropical environment of Bangladesh. AB - Soil-to-plant transfer factors (TF) of 137Cs and 90Sr have been determined for different plants/crops, such as rice, beans, peanuts, pineapple, cabbage, tomato, spinach and grass. They were obtained from radioisotope experiments on plants grown in pots under outdoor ambient tropical conditions for three growing seasons (1994-1996). In the case of 137Cs and concerning the above mentioned plants/crops, the average TFs were found to be 0.28, 0.25, 0.77, 0.19, 0.23, 0.28, 0.59 and 0.18, respectively. In the case of 90Sr, the average TFs were found to be 0.82, 0.51, 0.20, 0.82, 0.69, 0.59, 0.91 and 0.84, respectively. A minor seasonal variation was observed. This study provides a database of TFs for tropical environments to be used, e.g., for radiological safety assessment models. PMID- 9728746 TI - Generic relationship between calcium intake and radiostrontium transfer to the milk of dairy ruminants. AB - The hypothesis is tested that there is a generic relationship between the calcium intake and the transfer of radiostrontium to milk which can be used for all dairy ruminants. In addition to the daily calcium intake, the relationship also requires values for the strontium to calcium observed ratio, which describes the discrimination in transfer of the two elements to milk (a value of 0.11 is used), and the calcium concentration in milk. The relationship had previously only been validated for dairy cattle as there were insufficient data for other ruminant species. Here, we present recently available data for dairy goats, and also a limited amount of data for sheep derived from the literature. From the comparison between these data and predicted values, we conclude that it is possible to derive a generic model of the transfer of radiostrontium to the milk of dairy ruminants. PMID- 9728747 TI - Radioresistant cell strain of human fibrosarcoma cells obtained after long-term exposure to x-rays. AB - A radioresistant cell strain from human fibrosarcoma HT 1080 has been obtained after prolonged exposure to x-rays for 7 months (2 Gy per day, 5 days per week). This new strain, HT1080R, differs from HT1080 in a significantly increased ability of clonogenical survival, with coefficient alpha decreasing from 0.161 to 0.123 Gy(-1) and coefficient beta decreasing from 0.0950 to 0.0565 Gy(-2). Furthermore, the radioresistance of HT1080R proved to be stable in long-term passaged cultures as well as in frozen samples. Differences between the two cell lines are also observed in the G-banded karyotype; the new cell line shows monosomy of chromosome 17 and loss of 5p+ and 11q+ present in the parental cells. These data suggest that the radioresistance may have been caused by radiation induced cell mutation and that the resistant cells may have been selected by repeated irradiations. In order to characterize this new strain, the ability of the cells to rejoin DNA double-strand breaks, the cell cycle distribution and the amount of apoptosis after irradiation have been estimated; however, no differences are observed between these two cell strains. Although the mechanism of the elevated radioresistance remains unknown, this pair of cell strains can provide a new model system for further investigations with regard to the mechanisms of cellular radioresistance. The results also show that any type of irradiation similar to the schedules used in radiotherapy can lead to the formation and selection of more radioresistant cell clones in vitro, a phenomenon with possible implications for radiotherapy. PMID- 9728748 TI - Scanning and transmission electron microscopical evidence of the capacity of diatoms to penetrate the alveolo-capillary barrier in drowning. AB - The diagnostic value of diatom analysis for drowning is considered to be one of the most controversial arguments in forensic medicine. However, the theoretical assumption of the method, i.e. the capacity of diatoms to penetrate the alveolo capillary barrier during drowning, has never been addressed. Using scanning (SEM) and transmission electron microscopy (TEM), we have investigated the interaction of a natural population of diatoms and an unialgal culture of Phaeodactylum tricornutum (PT) with the alveolo-capillary barrier in an experimental model of drowning. The SEM analysis allowed the identification of several diatom species along the whole airways and their close interaction with the alveolar wall, but was poorly informative about the effective penetration of diatoms into pulmonary vessels. The TEM analysis was more informative and allowed a precise identification of the PT cells in alveolar spaces and to detect their phagocytosis by alveolar macrophages. PT penetrated into the pulmonary vessels through the thinnest portions of the alveolo-capillary barrier and through the interstitial spaces and were identified in pulmonary capillaries and venules. The morphological demonstration of the capacity of diatoms to penetrate the alveolo capillary barrier is a step forward in assessing the potentiality, reliability and limitations of diatom analysis on a new basis as a tool for the diagnosis of drowning. PMID- 9728749 TI - The diagnosis of amniotic fluid embolism: an immunohistochemical study for the quantification of pulmonary mast cell tryptase. AB - Recent clinical articles have suggested that amniotic fluid embolism (AFE) may be the result of anaphylactic reactions to fetal antigens and that the major part of this clinical syndrome is the result of mast cell degranulation and of the release of histamine, tryptase and other mediators. Tryptase, a neutral protease, is known to be the dominant protein component of the secretory granules of T and TC mast cells. In this paper we have examined the presence and the pulmonary distribution of mast cell tryptase utilizing specific immunohistochemical studies and morphometric evaluation in six cases of fatal amniotic fluid embolism compared to six subjects who died following anaphylactic shock and two control groups (five and six cases respectively) of traumatic death. The results demonstrate a numerical increase of pulmonary mast cells in the subjects who died of AFE (average cell number 54.095) with values corresponding to those encountered in cases of death due to anaphylactic shock (average cell number 51.378) compared with that of the traumatic control groups (average cell number 24.477 and 9.995 respectively). These results can shed light on additional criteria for the diagnosis of amniotic fluid embolism. PMID- 9728750 TI - Sequence variation of a hypervariable short tandem repeat at the D1S1656 locus. AB - A short tandem repeat at the D1S1656 locus was sequenced in 45 selected alleles and 13 different alleles were found which were designated according to the total number of repeats. This STR is a compound hypervariable STR consisting of blocks of (TAGR) repeats with a basic sequence structure (TAGA)4(TGA)0-1(TAGA)6 16(TAGG)0-1(TG)5. The presence of a TGA, probably due to an A deletion in the fifth TAGA repeat leads to intermediate a.3 alleles. Population data showed that this is a highly polymorphic STR with a heterozygosity of more than 0.89. This fact together with its simple structure and small size (129-168 bp) makes this STR one of the most interesting DNA polymorphisms for forensic and genetic purposes. PMID- 9728751 TI - Analysis of nine short tandem repeat (STR) loci in the Slovenian population. AB - A polymerase chain reaction (PCR)-based short tandem repeat (STR) system consisting of nine loci has recently been introduced in Slovenia for use in routine forensic identity testing. Fluorescently labelled PCR products were analysed using an ABI PRISM 310 Genetic Analyzer. The STR loci analysed exhibit between 6 and 14 observed alleles per locus and have a combined matching probability of 2.3 x 10(-10). PMID- 9728752 TI - Time-dependent expression of interleukin-10 (IL-10) mRNA during the early phase of skin wound healing as a possible indicator of wound vitality. AB - This study was performed to clarify whether interleukin-10 messenger RNA (IL-10 mRNA) could be a possible indicator for the distinction between intravital wounds and postmortem damage. After incision, mice were sacrificed from 0 to 180 min. The initial amount of IL-10 mRNA in each skin specimen was evaluated using the reverse transcriptase-polymerase chain reaction (RT-PCR). After 15 min there was a rapid increase in IL-10 mRNA which peaked at 60 min. A significant increase in IL-10 mRNA occurred between 30 and 180 min. During the 5 day postmortem interval the increase in time-dependent IL-10 mRNA expression was maintained and no significant increase in IL-10 mRNA expression occurred in the postmortem control. The increased expression of IL-10 mRNA could be considered a vital reaction in skin specimens with postmortem change. This study demonstrated the possible use of mRNA analysis for forensic wound examination because mRNA was detectable by RT PCR over a longer postmortem time course. PMID- 9728753 TI - Lipid peroxidation in lung tissue after chest trauma and correlation with the duration of the post-trauma survival period. AB - Blunt chest trauma is a frequently encountered cause of death in forensic pathology and one of the organs most likely to be affected are the lungs. Assuming that the victim survives the initial trauma, reperfusion processes take place, free radicals are formed and lipid peroxidation occurs. The aim of this study was to ascertain whether the length of the survival time is correlated with the extent of lipid peroxidation in the lung tissue following such ischaemia reperfusion processes. A study of 470 samples taken from all five pulmonary lobes from 94 cadavers was carried out. Cases were allocated to different groups according to whether there was chest trauma and/or a known survival period. Lipid peroxidation was investigated by determining malondialdehyde (MDA) levels. The lowest mean level of peroxidation was found in control cases showing no evidence of chest trauma at autopsy and no apparent survival period. Our results suggest that the level of lipid peroxidation in lung tissue can be a reliable indicator of survival processes. PMID- 9728754 TI - Performance enhancing drugs (doping agents) and sudden death--a case report and review of the literature. AB - The case of sudden cardiac death of a 23-year-old body builder who used anabolic steroids combined with other performance enhancing drugs is reported. Postmortem investigations revealed cardiac hypertrophy, acute cellular necrosis and interstitial fibrosis of the myocardium. The side-effects and interactions of the substances used are discussed. PMID- 9728755 TI - A case of fatal benzalkonium chloride poisoning. AB - Five elderly persons with senile dementia accidentally ingested Hoesmin, a 10% aqueous solution of benzalkonium chloride (BAC). The condition of one patient, an 84-year-old woman whose lips and oral cavity became erythematous, gradually deteriorated. Although gastric lavage was performed, the patient died 3 h after ingestion of Hoesmin. Autopsy revealed corrosive changes of the mucosal surfaces of the tongue, pharynx, larynx, esophagus and stomach which may have come in contact with BAC. In addition, BAC was detected in the serum. We conclude that the patient died of BAC poisoning. Fatal BAC poisoning is rare and autopsy findings in only a few cases of BAC poisoning have been reported. Our findings emphasize the risk of oral ingestion of BAC. PMID- 9728756 TI - New reference allelic ladders to improve allelic designation in a multiplex STR system. AB - This paper reports the composition of a new reference allelic ladder mixture for use with a multiplex DNA profiling system consisting of six short tandem repeat loci. The loci included in this mixture are HUMTH01, D21S11, D18S51, D8S1179, HUMVWAF31/A, HUMFIBRA/FGA and an amelogenin sex test. Sequence analysis of individual ladder alleles was carried out and allelic designations made in accordance with the recommendations of the International Society of Forensic Haemogenetics (1992; 1994). A series of rare alleles which increase the range of alleles previously reported were identified. By including some of the rare alleles into the ladder marker system, we have significantly improved the ability to identify new alleles in unknown samples. PMID- 9728757 TI - D1S80 allele frequencies in Hasidic and non-Hasidic New York City Jewish populations. AB - Allele frequencies were determined for the VNTR locus D1S80 in Hasidic and non Hasidic Ashkenazi New York City Jewish subpopulations. Samples were amplified via the polymerase chain reaction and underwent genotyping using polyacrylamide gel electrophoresis. In the Hasidic population 14 alleles were observed as opposed to 19 alleles in the non-Hasidic community. Both populations were tested for Hardy Weinberg equilibrium. The frequency data obtained can be used for comparison to other populations and for allele and genotype frequency estimates in genetic marker profiling of evidentiary specimens. PMID- 9728758 TI - Distribution of D1S80 alleles in the Jordanian population. AB - This study demonstrates that the locus D1S80 is highly polymorphic, with 24 different alleles and 66 genotypes in 215 Jordanians. This data set conforms to Hardy-Weinberg expectations(HWE). PMID- 9728759 TI - Is the incidence of SIDS increasing in Asia? AB - The incidence of SIDS in western countries has decreased since the late 1980s after active SIDS prevention campaigns. By contrast, both Taiwan and Japan have reported an increase in incidences. In this report data from England, Wales, Scotland, Northern Ireland, Japan and Hong Kong are analysed. Regression coefficient and linearity of regression tests were used to determine if there was any significant increase or decrease in post-neonatal mortality (PNNM), SIDS mortality and mortality among infants 1-5 months old. A statistically significant decrease in SIDS was recognized in England, Wales, Scotland and Northern Ireland, whereas statistically significant increases were noted for Japan and Hong Kong. However in Japan and Hong Kong both the PNNM and mortality of infants between 1-5 months in age fell significantly, suggesting that the increased SIDS rates could be due to a change in diagnostic labelling. Further study will be required to determine whether this increase in the incidence of SIDS is genuine or only an artifact. PMID- 9728760 TI - Population study of the short tandem repeat polymorphisms HumTH01, HumvWA31, HumFESFPS and HumF13A01 in Sicily (Southern Italy). AB - Population genetic studies were carried out on randomly selected and unrelated healthy individuals from Sicily (n = 140-150 individuals) using the short tandem repeat (STR) systems HumTH01, HumvWA31, HumFESFPS and HumF13A01. After vertical electrophoresis on polyacrylamide denaturing gels 6 alleles could be identified for TH01, 9 for vWA31, 7 for FESFPS and 11 for F13A01. No significant deviations from Hardy-Weinberg were observed. PMID- 9728761 TI - Potential DNA mixtures introduced through kissing. AB - The use of saliva samples is an alternative to blood samples when a large number of control samples are to be compared by DNA investigations. The most convenient and safe method is by using cotton wool swabs. In this investigation the average DNA content of saliva samples taken by three different sampling techniques (i. e. cotton wool swab, filter paper, liquid saliva) was compared. In addition the possibility of a DNA mixture of saliva samples after intensive kissing was investigated by taking samples from voluntary pairs. Mixed STR patterns were found in five samples but restricted to the first sampling after kissing within max. 60 s. PMID- 9728762 TI - Altered expression of immune-related antigens by neuronophages does not improve neuronal survival after severe lesion of the facial nerve in rats. AB - Injection of Fluoro-Gold (FG) into the whiskerpad muscles of rats yields a permanent retrograde labeling of motoneurons in the facial nucleus. Following subsequent resection of 10 mm of the facial nerve, one-third of the facial motoneurons die and the microglia phagocytize the dead FG-labeled neurons, take up FG, and get labeled in vivo. The resulting identification of all FG-labeled cells allows long-term comparative investigations on the behavior of neuronophages. In this study, we used two groups of rats to test whether the quantified expression of five immune-related antigens by neuronophages was related to quantified decline in neuron number (counts after immunostaining for neuron-specific enolase) 3 to 224 days after resection of the facial nerve. Rats of the first group received standard food and those of the second group, pellets containing 1,000 ppm of the calcium channel blocker nimodipine. Image analysis of the number of FG-containing cells and the number and projection area of immunopositive neuronophages in serial sections for each antigen showed that nimodipine significantly attenuated the immunostaining for CR3, MHC class I, and class II antigens (monoclonal antibodies [MAbs] OX-42, OX-18, and OX-6); enhanced the expression of monocyte-macrophage-specific antigen (MAb ED1); and did not change the expression of rat macrophage differentiation antigen (MAb ED2). The altered expressions, however, had no effect on the loss of motoneurons in the lesioned facial nucleus. We conclude that the degree of expression of immune related antigens by neuronophages has no influence on the delayed neuronal cell death induced by permanent target deprivation. PMID- 9728763 TI - N-acetylaspartylglutamate activates cyclic AMP-coupled metabotropic glutamate receptors in cerebellar astrocytes. AB - N-Acetylaspartylglutamate (NAAG) is the most prevalent peptide in the mammalian nervous system. NAAG meets the traditional criteria of a neurotransmitter, including localization in synaptic vesicles, depolarization-induced release, low potency activation of some N-methyl-D-aspartate receptors, and highly selective activation of a cAMP-coupled metabotropic glutamate receptor (mGluR) with potency approaching that of glutamate. The peptide is present in cultured cortical glia in high concentration and is hydrolyzed by cell surface peptidase activity. In the present work, we tested the hypothesis that NAAG selectively activates a subclass of metabotropic receptors on cultured rat cerebellar glia, primarily astrocytes. These glial cells express mRNA for mGluR subtypes 1, 3, 4, 5, 6, 7, and 8. We were not able to detect message for mGluR2 in these cells using the reverse transcriptase-polymerase chain reaction. Cerebellar glia responded to NAAG, glutamate, and trans-ACPD with a decrease in forskolin-stimulated cAMP formation. AP4, an agonist of the group III receptors mGluR4, mGluR6, mGluR7, and mGluR8, had little or no effect on stimulated cAMP levels. Treatment with low micromolar NAAG significantly increased uptake of radioactive thymidine by cultured astrocytes through activation of metabotropic glutamate receptors. Antagonists of group II mGluRs prevented the decrease in cAMP and the increase in uptake of thymidine by NAAG. Cultured cerebellar astrocytes expressed 20 pmol NAAG per mg protein, a value that is at least 30-fold lower than that expressed by mixed glial cultures prepared from mouse cortex. We conclude that cerebellar astrocytes respond to NAAG via the mGluR3 receptor and that the peptide may selectively activate this receptor subtype in astrocytes following release from neurons or glia. PMID- 9728764 TI - Identification of CSF-1 as a brain macrophage migratory activity produced by astrocytes. AB - Intraparenchymal migration of macrophages occurs in the CNS during development or as a consequence of tissue injuries. In the present study, we have shown, by using an in vitro chemotaxis assay, that cultured rat astrocytes obtained from the developing cerebral cortex and striatum produce soluble factors, which attract purified brain macrophages. The effect of astrocyte-derived factors on macrophages was strongly reduced in the presence of antibodies neutralizing colony-stimulating factor 1 (CSF-1, also called M-CSF), and recombinant CSF-1 was found to act as a chemotactic agent on brain macrophages. Synthesis of CSF-1 by cultured astrocytes was confirmed by northern detection of CSF-1 transcripts. In contrast, the CSF-1 gene was not expressed by cultured neurons from the cerebral cortex and striatum or by the brain macrophage population responsive to CSF-1 gradient. ELISA detection of CSF-1 in tissue extracts revealed the occurrence of this cytokine in the rat cerebral cortex during postnatal development and in adults. Altogether, our results demonstrate that astrocytes, through CSF-1 secretion, can trigger the polarized migration of brain macrophages and suggest a new mechanism which could regulate the locomotion of these cells in the cerebral cortex during ontogenesis or following lesions. PMID- 9728765 TI - Localization of p75 neurotrophin receptor in the retina of the adult SD rat: an immunocytochemical study at light and electron microscopic levels. AB - The low-affinity neurotrophin receptor p75(NTR), or p75, is a 75-kDa cell surface glycoprotein that binds all neurotrophins with similar affinity and is thought to help to ensure the specificity of each neurotrophin. In order to better understand the role of p75 and how it is involved in the neurotrophic effects in the retina, we have examined its cellular localization in the adult rat retina by immunocytochemistry at both light and electron microscopic levels. The similarity between the staining pattern of p75 and that of the distribution of Muller cell processes, as marked by antibodies against S-100 and vimentin, suggests that p75 may be on the Muller cell processes and not on the retinal ganglion cells (RGCs) as previously reported. The failure to detect p75 immunoreactivity on Fluoro-Gold retrogradely labeled RGCs in the radially sectioned retinae also indicates that it is not expressed on RGCs. The results from the light microscopic immunohistochemical studies are supported at the ultrastructural level, showing that p75-immunopositive staining is localized on Muller cell processes and not on RGC bodies. Muller cell processes not only form the inner limiting membrane but also partially wrap around the RGC bodies. Our results lead us to conclude that the previously reported immunopositive staining of p75 on RGCs might belong to the surrounding Miller cell processes. Thus, the pathway of neurotrophic effects on RGCs might be, at least partially, through a glial-neuronal pathway rather than on RGCs directly. PMID- 9728766 TI - Induction of resting microglia in culture medium devoid of glycine and serine. AB - Cultured microglial cells usually exhibit ameboid morphology and peripheral macrophage-like properties, which are distinct from those observed in the normal mature brain. This might be caused by the inappropriate culture of microglial cells in high concentrations (approximately 200-400 microM) of Gly and Ser, although the concentrations of the amino acids in extracellular spaces of the brain parenchyma are quite low (approximately 5 microM). In the present study, we focused on the concentration-dependent effects of glycine (Gly) and serine (Ser) on microglial morphology and function. Under Gly/Ser-free and serum-free condition, the majority of rat microglial cells displayed round morphology, whereas in the presence of 5 microM Gly and 25 microM Ser, which correspond to the concentrations of Gly and Ser in the cerebrospinal fluid, they extended multiple branched processes and formed clusters of rough endoplasmic reticulum. On the other hand, Gly and Ser did not affect morphology of astrocytes. The viability of microglia was not affected by the changes in the concentrations of Gly and Ser. Metabolic activity, activities of acid phosphatase and inducible nitric oxide synthase, and superoxide anion (O2-) generation were all strongly suppressed in Gly/Ser-free medium or in medium containing physiological concentrations of both amino acids. Such activities were all enhanced in harmony with increases in the concentrations of Gly and Ser. Thus, microglial cells cultured in Gly/Ser-free medium, even though exhibiting ameboid morphology, appears to be in the functionally resting state. Furthermore, once the resting state was achieved, the microglial cells remained inactive even after the subsequent 24 h culture in serum-supplemented medium containing 400 microM of both amino acids. The medium conditioned by microglial cells that were cultured in the presence of 400 microM of Gly and Ser was toxic to cortical neurons, whereas the microglia-conditioned medium obtained in the absence of both amino acids facilitated the survival of cortical neurons. Therefore, microglial cells in the resting state, which was induced in the Gly/Ser-free condition, are likely to support neurons. Microglial cells could ramify on glass coverslips coated with astrocyte-derived extracellular matrix or on coverslips coated thinly with fibronectin and/or laminin even under the Gly/Ser-free condition. The ramified cells as induced in this way kept suppressed O2- generating activity. These findings suggest that resting ramified microglial cells with a neurotrophic activity can be induced with the combination of Gly/Ser-free medium and small amounts of extracellular matrix proteins, and that the resting state is rather stable. PMID- 9728767 TI - Effects of hypoxia on glial cell expression of angiogenesis-regulating factors VEGF and TGF-beta. AB - Perivascular glial cells are thought to be involved in physiologic vascularization and also in pathologic angiogenesis in the central nervous system. We have previously shown that astrocytes are a source of transforming growth factor-beta (TGF-beta) and another inhibiting factor, which block endothelial cell growth and induce their apoptosis. Astroglia are also known to express vascular endothelial growth factor (VEGF), which is up-regulated during hypoxia. Here we demonstrate the effects of hypoxia on the expression of both TGF beta and VEGF by retinal glial cells. Muller cells isolated from rat retina were incubated under hypoxia or normoxia and the resulting conditioned media (H-MCM and N-MCM) were assayed for their effects on growth of bovine retinal capillary endothelial (BRE) and the TGF-beta-sensitive mink lung epithelial CCL cells. The expression and quantities of VEGF and TGF-beta (active vs. latent form) were determined by immuno-adsorption, Western or Northern blotting, and ELISA. N-MCM stimulated BRE cell growth by twofold but inhibited CCL cells under similar assay conditions, whereas H-MCM had a weak stimulating effect on BRE and substantial inhibitory activity on CCL cells. Adsorption of MCM by specific antibodies as well as Western and Northern blot analysis indicated that stimulating and inhibitory activities of MCM are due to the presence of VEGF and TGF-beta, respectively. ELISA revealed that the hypoxia condition converts latent TGF-beta into its active form. In N-MCM, TGF-beta is found predominantly in the latent form, but in hypoxia MCM it is mainly active. Furthermore, it was found that treatment of Muller cells with exogenous TGF-beta under either hypoxia or normoxia increases VEGF expression in a time- and dose-dependent fashion. TGF beta activation may, therefore, be prerequisite for hypoxia-induced up-regulation of VEGF and stimulation of angiogenesis in vivo. PMID- 9728768 TI - Cotranslational integration of myelin proteolipid protein (PLP) into the membrane of endoplasmic reticulum: analysis of topology by glycosylation scanning and protease domain protection assay. AB - The four transmembrane domain topology of the proteolipid protein (PLP) in the myelin membrane of the central nervous system (CNS) has been further substantiated by biochemical studies. We have analyzed the cotranslational polytopic integration of nascent PLP during protein synthesis into the membrane of the endoplasmic reticulum (ER) on two routes. Consensus sequences for N glycosylation were introduced by site directed mutagenesis into the PLP sequence as reporter sites, which upon glycosylation monitor the intraluminal location of the respective domains corresponding to the extracellular side of the plasma membrane. Single, double, and triple mutant cDNAs were constructed for transcription/translation in vitro in the presence of ER-membranes. The glycosylation pattern of the translation products revealed that hydrophilic extramembrane regions 2 and 4 (EMR2/EMR4) and EMR3 of PLP are exposed on opposite sides of the ER membrane. Their localization either at the cytosolic or luminal side of the ER membrane leads to two different topologies. The two modes of membrane integration during in vitro cotranslational translocation were confirmed by protease protection assays with wild-type and truncated PLP polypeptides with either one, two, or three putative transmembrane domains integrated into the ER membrane. The fragment pattern of the [35S]methionine- or [3H]leucine-labeled polypeptides revealed that EMR3 and EMR4 were exposed with opposite orientation either on the cytosolic or luminal side of the ER membrane supporting the 4 transmembrane helix (TMH) N(in) model with the N and C termini on the cytoplasmic side, as established for the myelin membrane (plasma membrane); the other inversely integrated PLP constructs indicate the 4-TMH-Nout profile. These results are discussed with regard to the PLP biogenesis and the plasma membrane topology in PLP-expressing cells. PMID- 9728769 TI - Migration arrest in glioma cells is dependent on the alphav integrin subunit. AB - Local invasion is a hallmark of gliomas. Infiltrating tumor cells establish sites of persistent and recurrent lesions that ultimately prove fatal. Determinants of glioma cell migration include integrins and their ligands within the matrix. In contrast to the response to other matrix proteins, glioma cells migrating on tenascin do not follow a characteristic dose-dependent pattern. For two of four glioma cell lines tested, tenascin acts as both a permissive and a nonpermissive motility substrate, i.e., low densities of tenascin are permissive substrates, whereas high densities are nonpermissive. Specific antisense oligonucleotides directed at the alpha(v) integrin subunit effectively suppress the anti-migratory phenotype of glioma cells at high tenascin densities. The two cell lines that fail to demonstrate this unusual biphasic pattern do not endogenously express the alpha(v) subunit, whereas the cell lines for which high densities of tenascin are anti-migratory are found to express alpha(v). We conclude that loss of the alpha(v) integrin subunit may be associated with the invasive behavior of gliomas, along vascular channels that express tenascin. PMID- 9728770 TI - Allograft-inflammatory factor-1 in rat experimental autoimmune encephalomyelitis, neuritis, and uveitis: expression by activated macrophages and microglial cells. AB - Allograft inflammatory factor-1 (AIF-1) is a Ca2+ binding peptide expressed predominantly by activated monocytes. In order to investigate the role of AIF-1 in autoimmune lesions of the rat nervous system, we have used a synthetic gene to express AIF-1 in E. coli and have produced monoclonal antibodies against AIF-1. AIF-1 was localized to monocytes/macrophages with rather selective staining of a minor rat monocyte subpopulation of lymphoid tissue. We then investigated expression of AIF-1 in experimental autoimmune encephalomyelitis (EAE), neuritis (EAN), and uveitis (EAU). Within the local inflammatory lesions, infiltrating macrophages are prominently stained. In the diseased brain, AIF-1-positive microglial cells are not only found in the direct vicinity of the infiltrate, but widespread activation is seen in the parenchyma. This is the first demonstration that AIF-1 is present in autoimmune lesions. Immunostaining of microglial cells is noteworthy, as these cells are strategically placed regulatory elements of CNS immunosurveillance. Thus, AIF-1 might be a valuable marker to dissect the local monocyte heterogeneity in autoimmune disease. PMID- 9728771 TI - Enhanced cellular glutathione peroxidase immunoreactivity in activated astrocytes and in microglia during excitotoxin induced neurodegeneration. AB - The cellular distribution pattern of cellular glutathione peroxidase (GPx) was analyzed immunocytochemically in the normal rat central nervous system (CNS) and following exposure to the excitotoxin quinolinic acid (Quin). In the normal CNS, GPx was localized predominantly to microglia, identified by staining with isolectin B4 or the monoclonal antibody OX-42. Three days after intrastriatal administration of 90 pmoles Quin, GPx immunoreactivity was increased in activated microglia and also in astrocytes costained for glial fibrillary acidic protein (GFAP). Whereas GPx-positive astrocytes were seen predominantly in the environment of the lesion, most intensive immunoreactivity was located in globular-shaped microglia in the lesion core. GPx staining was generally low in neurons and failed to increase its intensity after lesion. In the case of excitotoxin-induced generation of oxygen-derived free radicals, the elevation of GPx levels in microglia, and likewise in activated astroglia, may provide an important mechanism to withstand oxidative stress. PMID- 9728773 TI - Audiometry in general practice: validation of a pragmatic pure-tone audiometry method. AB - The aim of this study was to validate the results of diagnostic pure-tone audiometry performed in a typical practice setting by comparing with test results obtained in a standardized audiological setting in accordance with the ISO standards. In a single-blinded crossover design, 119 persons were tested (0.25-8 kHz) in both settings. The mean deviations as a function of frequency were in the order of less than 2 dB (0.5-4 kHz) and otherwise up to 4 dB; the practice setting representing the poorer thresholds. The validity of the practice audiometry at three criteria of hearing impairment (0.5-4 kHz) was characterized by sensitivity (82-100%), specificity (95-99%); positive predictive values (75 90%) and negative predictive values (98-100%) focusing on the better ear. It is concluded that pure-tone audiometry of appropriate validity can be performed in general practice and that it is useful in selecting patients with no need of further audiological examination. Guidelines are needed. PMID- 9728772 TI - Audiological disturbances caused by long-term exposure to industrial solvents. Relation to the diagnosis of toxic encephalopathy. AB - Sixty workers, consecutively admitted due to suspicion of solvent-induced chronic toxic encephalopathy (CTE), were investigated with pure-tone audiometry, determination of speech recognition of monosyllabic words and distorted speech and cortical response audiometry (CRA). Eighteen workers not exposed to occupational solvents and noise were also investigated. The scores in the distorted speech test were significantly lower and the CRA latencies were significantly longer in the solvent group than in the control group. There was no difference between the groups in the pure-tone and monosyllabic speech recognition tests. In the solvent group, 19 subjects had one or several pathological audiological test results (values exceeding the mean result of the control group by 2 SD). Independently of the audiological examination all the workers in the solvent group underwent the traditional clinical assessment of CTE, which is based on symptoms, history of exposure, clinical neurological examination and a neuropsychological investigation. They were classified in three groups--CTE, incipient CTE and non-CTE. There was no correlation between these groups and the audiological test results. A previous report on vestibular pathology in the same group of subjects and the present investigation on hearing deficits suggest that long-term exposure to solvents causes disturbances of the central pathways in the otovestibular system. Hitherto, no attention has been paid to these disturbances in the definition of the CTE syndrome. PMID- 9728774 TI - Self-reported need of information, counselling and education: needs and interests of re-applicants. AB - An interview study was performed to establish and evaluate the need for education and counselling of hearing aid users (HA users) in order to review present procedures of information and potential referral to the educational sector. The study comprises 102 consecutively selected hearing-impaired adults (>18 years), the only inclusion criterion being experienced HA user. The interview took place immediately after the patient's latest HA fitting appointment and consisted of 24 structured and closed questions. The findings indicate that a significant proportion of HA users, especially the elderly, do not experience a need for or show an interest in further education and counselling concerning their hearing handicap. It is concluded that the present organization of services meets the demands of a majority of re-applicants. PMID- 9728775 TI - Aural rehabilitation in the elderly: supply of hearing aids related to measured need and self-assessed hearing problems. AB - Three age cohorts of elderly persons in Goteborg (70, 75 and 88 years of age) were studied regarding hearing aid (HA) rehabilitation, on the one hand, and measured and self-assessed hearing, on the other. The participants, 615 in number, were representative of their ages and were selected from a geriatric population study. At age 70, 12% of the participants had been equipped with HAs. At age 75, the corresponding figure was 14% and at age 88, 32%. The correlations between self-assessed and audiometrically measured hearing were reasonably high (r = 0.5-0.7). According to the result for the self-assessed measure, we estimate that elderly persons with pure-tone averages (PTAs) at 30 dB HL (0.5-4 kHz, better ear) are in need of aural rehabilitation. Nevertheless, few subjects with PTAs between 30 and 49 dB HL have been equipped with HAs. At age 88, almost 20% of those with pronounced problems had no HA. Very few participants with no documented hearing problems for aural rehabilitation had been equipped with HAs. PMID- 9728776 TI - Long-term test-retest reliability of category loudness scaling in normal-hearing subjects using pure-tone stimuli. AB - The present study investigates the test-retest reliability of category loudness scaling with pure tones for each of the scaling categories: 'very soft', 'soft', 'OK', 'loud', 'very loud' and 'too loud' at the audiometric frequencies 0.5, 1 kHz, 2 kHz and 4 kHz. Category loudness scaling at two sessions separated by between 1 and 4 weeks was obtained from 16 normal-hearing subjects who all had normal otoscopy, present acoustic reflexes at audiometric frequencies 0.5-4 kHz and middle ear pressure within +/-50 daPa. Intra-subject between-session reliability was found not to be frequency dependent, and comparison with other studies revealed that reliability is not dependent on the applied stimulus signal. Test-retest reliability varied between the different categories: In the categories 'very soft', 'loud', 'very loud' and 'too loud' the test reliability is in the same range as found for hearing thresholds determination, whereas for the 'soft' and 'OK' categories it is poorer. The greater uncertainty for intermediate levels should be considered when using category loudness scaling, e.g. for calculating hearing aid parameters. PMID- 9728777 TI - Average thresholds in the 8 to 20 kHz range in young adults. AB - The sound-pressure level thresholds in the extended high-frequency range (8 to 20 kHz) were measured in 25 non-hearing-impaired young adults from 20 to 29 years of age. The result was not unlike that obtained by previous investigators; the thresholds increased gradually as a function of frequency. However, two notable points were found: one that the threshold reached a plateau above 18 kHz, and the other that it decreased slightly at 12 kHz. As the subjects might respond to the low-frequency noise of the stimulus wave, the threshold became a plateau above 18 kHz. An acoustic resonance in the ear canal caused the threshold to decrease at 12 kHz. In clinical studies of extended high-frequency audiometry, the threshold data should be carefully evaluated above 18 kHz and at 12 kHz. PMID- 9728778 TI - Effect of probe frequency and gender on click-rate-induced facilitation of the acoustic reflex thresholds. AB - This study evaluated the effects of probe frequency and gender on the click-rate induced facilitation of the acoustic reflex thresholds (ARTs). ARTs were measured by delivering clicks at the repetition rates of 60, 120, 180, 240 and 300/sec. The probe tones were 226, 678 and 1000 Hz. Rate-induced facilitation (RIF) was calculated for each subject, for each of the probe tones, by subtracting the minimum ART from the maximum ART. The mixed MANOVA on the RIF revealed no significant main effects or interactions for gender or probe tone frequency. However, paired comparisons revealed that the RIF values obtained from male subjects with the 1000 Hz probe tones were significantly lower than those obtained from the female subjects with all the probe tone frequencies. The results suggest that RIF can be studied with higher probe tones, but gender differences need to be considered if the probe tone is 1000 Hz. PMID- 9728779 TI - A new more powerful bone-anchored hearing aid: first results. AB - In a pilot study seven subjects were asked to use a new more powerful bone anchored hearing aid, the BAHA cordelle, for 5 days. All seven had a moderate or severe sensorineural component in their hearing loss and had been using a BAHA superbass on a daily basis for more than 2 years. Free-field thresholds and speech recognition were measured with the BAHA superbass and with the BAHA cordelle. A questionnaire was also filled in to obtain subjective information about speech recognition with the BAHA superbass compared to the BAHA cordelle. Aided thresholds in the high frequency range with the BAHA cordelle were better than with the BAHA superbass. In six of the seven subjects there was significant improvement in the speech recognition score with the BAHA cordelle. These six subjects expressed preference for the BAHA cordelle. PMID- 9728780 TI - Behavioural observation audiometry in testing young hearing-impaired children. AB - The aim was to examine the accuracy of unconditioned behavioural observation audiometry (BOA) in predicting hearing acuity in children and the validity of test results at various frequencies. The study was designed to longitudinally compare each child's best BOA response level (at the age >12 months) with the conclusive pure-tone threshold of the better ear. The subjects were 119 children derived from a material of 353 children fitted with a hearing aid at Helsinki University Central Hospital. BOA was carried out on 119 children, 19 of whom did not respond to frequency-specific stimuli. The predictive power of BOA depended on the severity of hearing loss. At the hearing level of 30-39 dB, BOA registered 10-15 dB poorer levels than the pure-tone audiometry. The pure-tone averages (0.5, 1, 2 kHz) of 50-69 dB agreed best with the BOA responses. In severe impairments (more than 70 dB HL), the BOA registered too good hearing. Correlation of the results from the two modes to measure hearing level was highly significant (r = 0.71, p = 0.000), and the pure-tone hearing level agreed with that of BOA at the frequencies 0.5 to 4 kHz. Our results show that BOA averages < or =30 dB rarely indicate hearing loss demanding fitting of a hearing aid. PMID- 9728781 TI - Average thresholds in the 8 to 20 kHz range as a function of age. AB - The sound-pressure thresholds at the extended high frequencies of 8-20 kHz were measured for 65 normal subjects aged between 10 and 69 years. The results are not unlike those obtained by previous investigators. The thresholds increased gradually as a function of frequency, except around 12 kHz and above 19 kHz, and also as a function of age. To clarify the connections between threshold, frequency and age, we introduced the regression lines for the threshold by analysing two ranges of frequencies (8-10 kHz and 14-19 kHz) and determining their slopes and intercepts. The regression line analysis reveals that the thresholds at 8-10 kHz tend to increase more at higher frequencies as subject age increased above 30 to 39 years, and those at 14-19 kHz increase translationally with increase of age. Our results did not contradict earlier reports on the pathological changes of the inner ear. PMID- 9728782 TI - Accuracy of auscultation in the detection of haemopneumothorax. AB - OBJECTIVE: To assess the accuracy of auscultation in the detection of haemopneumothorax. DESIGN: Prospective study. SETTING: University hospital, Taiwan. PATIENTS: 148 patients with chest injuries admitted between July 1994 and August 1996. MAIN OUTCOME MEASURES: Correlation between the results of auscultation and radiographic findings in 148 patients with injuries to the chest. 83 (56%) had internal injuries, of whom 38 had pneumothoraces, 24 haemothoraces, and 21 haemopneumothoraces. RESULTS: Auscultation had a sensitivity of 84%, a specificity of 97%, an accuracy of 89% and a positive predictive value of 97% in the detection of these injuries. CONCLUSIONS: Auscultation is not as accurate as chest radiography. Chest tubes can be inserted before chest radiography in patients in whom auscultation has indicated an injury. A chest radiograph is essential in those patients with normal breath sounds to exclude a haemopneumothorax that had been missed by auscultation. PMID- 9728783 TI - Concentrations of cytokines in plasma of patients with large burns: their relation to time after injury, burn size, inflammatory variables, infection, and outcome. AB - OBJECTIVE: To monitor longitudinally the concentrations of cytokines in the plasma of patients with severe burns. DESIGN: Prospective open study. SETTING: Burns unit, university hospital, Norway. SUBJECTS: 27 patients (5 women and 22 men, mean age 37 (range 13-82) years). INTERVENTIONS: Measurement of plasma concentrations of interleukin-1beta(IL-1beta), interleukin-1 receptor antagonist (IL-1ra), interferon-7(IFN-gamma) and tumour necrosis factor-alpha (TNF-alpha) were measured by enzyme linked immunosorbent assays (ELISA). MAIN OUTCOME MEASURES: Changes in concentrations, and correlation with morbidity and mortality. RESULTS: The concentration of IL-1beta and IL-1ra were increased in all patients and highest at the time of admission. Initially there was little or no circulating IFN-gamma, but this increased from day 5-10 in all patients. Only 8/15 patients had transient increases in circulating TNF-alpha. Concentrations of IL-1ra correlated with total burn surface area (TBSA) and area of third degree burn, as well as with plasma concentrations of C - reactive protein (CRP). Concentrations of IL-1beta and IL-1ra were higher in patients who developed infective complications than in those who did not (interleukin-8 (IL-8) has previously been shown to follow this pattern as well). Patients who survived had significantly higher IL-1beta concentrations than those who died (13(1) compared with 3 (1) pg/ml, p = 0.005) CONCLUSION: There are significant time-dependent changes in plasma concentrations of IL-1beta, IL-1ra, IFN-gamma and TNF-alpha after serious burns. IL-1ra concentrations may be influenced by size of the burn and the acute phase response; IL-1beta, IL-1ra and IL-8 may have a role in the host's response to infection; and IL-1beta may influence outcome. PMID- 9728784 TI - Incidence and prevention of deep venous thrombosis occurring late after general surgery: randomised controlled study of prolonged thromboprophylaxis. AB - OBJECTIVE: To study the incidence of late deep venous thrombosis (DVT), and to evaluate a regimen of prolonged thromboprophylaxis after general surgery. DESIGN: Randomised, controlled, open trial, with blinded evaluation. SETTING: University hospital, Denmark. SUBJECTS: 176 consecutive patients undergoing major elective abdominal or non-cardiac thoracic operations, of whom 118 were eligible for evaluation. INTERVENTIONS: Thromboprophylaxis with a low-molecular-weight heparin, tinzaparin, given for four weeks (n = 58), compared with one week (control group, n = 60). MAIN OUTCOME MEASURES: Presence of DVT established by bilateral venography four weeks after the operation. RESULTS: The incidence of late DVT in the control group was 6/60 (10%, 95% confidence interval (CI) 4% to 21%). In the prophylaxis group it was 3/58 (5.2%, 95% CI 1% to 14%) (p = 0.49). CONCLUSION: Prolonged thromboprophylaxis had no significant effect on the incidence of DVT occurring late after general surgery. PMID- 9728785 TI - Value of serum thyroglobulin measurement in patients operating on for well differentiated thyroid carcinoma. AB - OBJECTIVE: To study the clinical relevance of measurements of serum thyroglobulin in patients undergoing total thyroidectomy for well differentiated thyroid cancer. DESIGN: Prospective study. SETTING: University hospital, Sweden. SUBJECTS: 194 patients operated on for well differentiated thyroid cancer from 1 January 1978 to 31 December 1992. INTERVENTIONS: All patients underwent total thyroidectomy by a standard technique, and were prospectively followed up at regular intervals by clinical examination and measurement of the serum thyroglobulin concentration. RESULTS: Six patients whose thyroglobulin concentrations after operation were low or undetectable had gradual increases leading to detection of recurrences that could be treated successfully. Six patients had gradual increases without detectable recurrences. In 12 patients thyroglobulin concentrations remained high after operation with no signs of thyroid tissue remaining, which we interpreted as persistence of the disease. No recurrence was found without an appreciable rise in the thyroglobulin concentration. Thyroglobulin antibodies were found in 81 (42%). CONCLUSION: Measurement of the serum thyroglobulin concentration is a valuable addition to the follow up of patients operated on for highly differentiated thyroid cancer. In many cases it is the first sign of recurrent disease, thereby facilitating early and successful treatment. PMID- 9728787 TI - Laparoscopic partial posterior fundoplication improves poor oesophageal contractility in patients with gastrooesophageal reflux disease. AB - OBJECTIVE: To investigate the effect of partial posterior fundoplication on oesophageal contractility in patients with gastrooesophageal reflux disease (GORD). DESIGN: Follow-up study with 6 months of survey. SETTING: University hospital, Austria. SUBJECTS: 24 consecutive patients with GORD and poor oesophageal contractility. INTERVENTIONS: Laparoscopic partial posterior fundoplication. Oesophageal contractility was assessed manometrically. MAIN OUTCOME MEASURES: Changes in measurements of mean contraction amplitudes in the distal oesophagus, the number of contractions with amplitudes of less than 30 mmHg, the number of interrupted and simultaneous contractions, and the total number of defective contractions. RESULTS: 16 of the patients (67%) complained of dysphagia preoperatively, and none postoperatively. The mean (SEM) amplitudes in the distal oesophagus improved significantly (level 442.4 mmHg (3.5) compared with 31.8 mmHg (3.3), p = 0.03, and level 5-45.7 mmHg (3.8) compared with 32.6 mmHg (3.7), p = 0.02), the number of contractions with amplitudes below 30 mmHg decreased (18.0% (5.7) compared with 38.3% (6.2), p = 0.02), as did the number of interrupted or defected contractions (11.5% (3.6) compared with 26.3% (5.5), p = 0.03, and 29.5% (6.5) compared with 66.6% (5.1), p < 0.0001 respectively). There was no significant effect on the number of simultaneous waves (p = 0.11). CONCLUSIONS: Partial posterior fundoplication improves poor oesophageal body motility. This results in improvement of preoperative dysphagia. PMID- 9728786 TI - Quality of life following repair of ruptured and elective abdominal aortic aneurysms. AB - OBJECTIVE: To find out whether patients undergoing repair of ruptured abdominal aortic aneurysms (AAA) had more emotional problems and limitations of lifestyle than those listed for elective resection. DESIGN: Retrospective study SETTING: Teaching hospital, Eire. SUBJETS: 28 patients, 14 in each group, matched for age, sex, duration of stay in the intensive care unit (ICU), hospital stay postoperatively, and length of time since operation. INTERVENTIONS: Application of structured questionnaire. MAIN OUTCOME MEASURES: Emotional problems, mobility, activities of daily living, ability to sleep, degree of psychological stress, presence of symptoms, and Rosser index to measure quality of life (QoL). RESULTS: There were no significant differences between the groups in any measure. CONCLUSIONS: Patients have few emotional difficulties or disturbances of QoL after emergency or elective repair of AAA. Survivors after repair of ruptured AAA can expect as good a quality of life as those operated on electively. These results support an aggressive approach to the treatment of ruptured AAA. PMID- 9728788 TI - Short and long term course of elderly patients with peptic ulcer bleeding- analysis of factors influencing fatal outcome. AB - OBJECTIVE: To study long and short term survival in patients aged 60 years or over admitted with a peptic ulcer bleeding and find out which factors influence outcome. DESIGN: Cohort study with matched controls. SETTING: Two emergency hospitals, Sweden PATIENTS: 676 of the 687 patients aged 60 years or over admitted to the two emergency hospitals serving Gothenburg, Sweden during 1989 1993 who fulfilled the diagnostic criteria and whose case notes were available for study. MAIN OUTCOME MEASURES: Seven year survival rates and odds ratios for risk factors based on multiple logistic regression analyses. RESULTS: 37 patients died and the timing was evenly distributed within the first 30 days of admission with a cumulated case-fatality rate of 5.5% at day 30. Mortality was increased among the patients compared with the control group during the subsequent years. Factors that influenced day 30 mortality were age and Forrest class. CONCLUSION: Mortality is increased among patients with peptic ulcer bleeding even long after the event. Old age and signs of recent haemorrhage increase the risk. PMID- 9728789 TI - Gastrointestinal leiomyosarcomas: experience of 14 cases and review of published reports. AB - OBJECTIVE: To present our experience of leiomyosarcoma of the gastrointestinal tract during a 10 year period and review similar publications. DESIGN: Retrospective study. SETTING: Teaching hospital, Greece. SUBJECTS: 14 patients who were treated for leiomyosarcoma between 1983 and 1993. INTERVENTIONS: Tumours were diagnosed by endoscopy, computed tomography, and contrast examination. All patients were treated by resection and no adjuvant treatment was given. MAIN OUTCOME MEASURES: Morbidity, mortality, and outcome. RESULTS: The diagnosis was made preoperatively in 10/14 patients: 4 of the tumours were locally advanced at the time of laparotomy, and two had liver metastases. Resection was potentially curative (all macroscopic tumour removed) in 12 patients (86%). Three patients died of recurrent disease at 5, 8 and 63 months, respectively. The mean follow up in the 11 survivors was 44 months (range 24-83). CONCLUSION: Resection is the treatment of choice for these tumours. A multicentre trial would be required to assess the effect of adjuvant chemotherapy. PMID- 9728790 TI - The effects of nitric oxide synthase inhibitors on acute necrotising pancreatitis in rats. AB - OBJECTIVE: To investigate the effects of the constitutive nitric oxide (NO) synthase inhibitor NG-nitro-L-arginine methyl ester (L-Name) and the inducible NO synthase inhibitor amino ethyl-isothiourea (AE-ITU) on acute necrotising pancreatitis (ANP) in rats. DESIGN: Laboratory study. SETTING: Medical school, Turkey. MAIN OUTCOME MEASURES: Morbidity, mortality, effects on activities of various enzymes, and histological picture. RESULTS: NO inhibitors increased the mortality (from 8/15, 53%, for ANP plus saline, to 12/15, 80%, for ANP plus L Name, and 13/15, 87%, for ANP plus AE-ITU and serum amylase activity, but had no effects on serum calcium concentrations, volume of ascites, or degree of pancreatic damage. L-Name caused hypoglycemia, and AE-ITU reduced activities of lactate dehydrogenase and liver transaminases, and concentrations of urea and creatinine. CONCLUSIONS: Constitutive NO synthase inhibition worsens the course of ANP, and inducible NO inhibition has beneficial effects on various systems. PMID- 9728791 TI - Placement of a peritoneal dialysis catheter with routine omentectomy--does it prevent obstruction of the catheter? AB - OBJECTIVE: To find out if routine omentectomy reduced the incidence of obstruction and other complications of catheters inserted for continuous ambulatory peritoneal dialysis (CAPD). DESIGN: Retrospective study. SETTING: Teaching hospital, Israel. SUBJECTS: 60 patients with end stage renal failure who needed catheters for CAPD. INTERVENTION: Routine omentectomy during insertion of the catheter, usually under local anaesthesia. MAIN OUTCOME MEASURES: Short and long term morbidity, and mortality. RESULTS: No patient died as a result of the procedure. The catheter obstructed in only one patient (2%) during a mean follow up period of 28 months (range 2-108), and 90% of the catheters survived one year. CONCLUSIONS: Routine omentectomy during insertion of a catheter for CAPD under local anaesthesia is safe and the incidence of obstruction is low. Prospective randomised studies are needed before it can be recommended as the procedure of choice. PMID- 9728792 TI - Massive liver haemorrhage and rupture caused by HELLP-syndrome treated by collagen fleeces coated with fibrin glue. PMID- 9728793 TI - Diffuse sclerosing variant of papillary carcinoma of the thyroid. PMID- 9728795 TI - Effects of O6-benzylguanine on growth and differentiation of P19 embryonic carcinoma cells treated with alkylating agents. AB - The purpose of this study was to evaluate the impact of modulating the repair of O6-alkylguanine adducts on the developmental toxicity of alkylating agents. Alkylating agents that have been shown to induce developmental toxicity following either in vitro or in vivo exposure were chosen for this investigation, and include methylnitrosourea (MNU), ethylnitrosourea (ENU), methyl methanesulfonate (MMS), and ethyl methanesulfonate (EMS). P19 cells are pluripotent murine embryonic carcinoma cells that can be induced by all trans retinoic acid (RA) to differentiate into cells that are biochemically and morphologically very similar to cells of the central nervous system. These cells are useful for studying the ability of chemicals to affect neuronal viability and differentiation. Neuronally differentiating P19 cells were pretreated with O6-benzylguanine (O6-Bg), a potent and specific inhibitor of the O6-alkylguanine-DNA-alkyltransferase (AT) protein that repairs lesions at the O6-position of guanine. In previous studies using micromass rat embryo midbrain cells, O6-Bg greatly potentiated the ability of MNU but not ENU to inhibit differentiation, and did not significantly alter the effects of either MNU or ENU on viability. In the P19 cells, we found that AT inhibition potentiated the effects of MMS, MNU, and EMS to inhibit both viability and differentiation. Additionally, AT inhibition had a much greater effect on toxicity of the methylating agents, as compared to the ethylating agents. These results suggest that O6-alkylguanine adducts can inhibit both viability and differentiation in P19 cells treated with alkylating agents. PMID- 9728794 TI - Possible rare involvement of O6-methylguanine formation as a significant mutational factor in mouse urinary bladder carcinogenesis models. AB - O6-methylguanine is known as one of the major premutagenic lesions in the human and rodent carcinogenesis process. O6-methylguanine-DNA methyltransferase (MGMT), which repairs methylated guanine bases, might prevent the G:C to A:T transition, and transgenic mice carrying this MGMT gene have been reported to be less sensitive to the carcinogenicity of certain alkylating agents. Here we utilized MGMT transgenic mice to assess the significance of O6-methylguanine formation during urinary bladder carcinogenesis. In experiment 1, 100 and 60 ppm N-butyl N(4-hydroxybutyl)nitrosamine was given for 20 weeks to transgenic and non transgenic mice in their drinking water. The incidences of urinary bladder carcinomas were not different between transgenic mice and non-transgenic mice. The mutational spectrum of the p53 gene was evaluated by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis and direct sequencing. The pattern of p53 mutations of transgenic and non-transgenic mice did not differ, and the frequencies of mutations were 40% and 42%, respectively. G:C to A:T transition mutations were particularly infrequent (1 of 14 mutations, 7%). In experiment 2, N-methyl-N-nitrosourea, which might induce O6-methylguanine in affected alleles, was given once a week, 3 times (total 5 mg) by direct instillation into the urinary bladder through an abdominal incision. No significant neoplastic lesions were detected, although the experiment was limited by severe toxicity of the treatment. p53 immunostaining was done and there was no difference in transgenic and non-transgenic mice. These results suggest that O6 methylguanine formation might not be a significant mutational factor in these mouse urinary bladder carcinogenesis models. PMID- 9728796 TI - Micronuclei analysis in lymphocytes of pesticide sprayers from Concepcion, Chile. AB - To estimate the genetic risk associated with pesticide exposure in a defined population, the frequency of micronuclei (MN) in peripheral blood lymphocytes from a group of 22 pesticide sprayers from Concepcion, Chile, occupationally exposed to pesticide mixtures was evaluated. After scoring 1,000 binucleated cells for each donor, no significant increases were observed either for the total number of MN or for binucleated cells with MN, when compared with a concurrent control population. In addition, when the effects of different confounding factors such as age, smoking, and drinking habits were considered, no significant effect was observed. Our conclusion is that, in this specific group of workers and under the particular conditions of exposure to pesticides, when evaluated by the micronucleus assay, no genetic risk was detected. PMID- 9728797 TI - Induction of DNA adducts in vivo in rat lung cells by fume condensates of roofing asphalt. AB - Many workers in the highway construction and roofing industries are potentially exposed to asphalt fumes. However, little is known regarding the carcinogenic hazards of these fumes to the exposed workers. Previous studies have shown that condensates of asphalt fumes are weakly mutagenic to bacteria and are capable of inducing micronucleus formation in cultured mammalian cells. In this study, the induction of DNA adducts in vivo in lung and white blood cells (WBCs) of rats by fume condensates of type I and type III roofing asphalts was investigated using 32P-postlabeling analysis. Male CD rats (3/group) received 3 intratracheal instillations of fume condensates in a 24-h period. DNA from both lung cells and WBCs were isolated and used to detect DNA adducts. Condensates of both roofing asphalt fumes caused DNA adduct formation in rat lung cells in a similar dose related manner. Under the conditions studied, however, neither type I nor type III fume condensate induced DNA adducts in WBCs. These results indicate that 1) condensates of fumes from both type I and type III have similar genotoxic activity, 2) chemicals in the condensates of roofing asphalt fumes can covalently bind to the DNA of rat lung cells, and 3) WBCs may not be a suitable surrogate for lung cells in DNA adduct studies of workers exposed to roofing asphalt fumes. PMID- 9728798 TI - Spontaneous and asbestos-induced transformation of mesothelial cells in vitro. AB - The processes of spontaneous and asbestos-induced transformation of rat mesothelium were studied using cell cultures obtained in the laboratory. The same changes in cell properties were established in both spontaneous and asbestos induced transformation: change in epidermal growth factor (EGF) response, in some cases appearance of fibroblast-like cells instead of polygonal ones, appearance of multilayer cell growth foci, and ability to grow in semisolid agar. The response to fibroblast growth factor, insulin-like growth factor 1, and insulin did not change during transformation as well as the P450 system activity measured by benz(a)pyrene (BP) and 7,12-dimethylbenzanthracene (DMBA) cytotoxicity. The asbestos-induced transformation began earlier than the spontaneous one. EGF began to stimulate mesothelium proliferation instead of its inhibition at 6-7 passages in the case of asbestos-induced transformation, whereas during spontaneous transformation this change began at 9-10 passages. Elongated rather than polygonal cells appeared at 10-11 instead of 17-18 passages (this morphological change did not take place at all lines studied). The ability to grow in semisolid agar was found at 14-16 passages with asbestos and at 22-24 passages without it. The results allow us to propose the necessity of a positive EGF response for mesothelial cell transformation and the similarity of mechanisms of spontaneous and asbestos-induced transformation. PMID- 9728800 TI - Transient response to multiple courses of pulse high-dose dexamethasone therapy in children with idiopathic thrombocytopenic purpura. PMID- 9728799 TI - A historical perspective and cellular regulation of megakaryocytopoiesis. AB - With the recent accumulation of information about megakaryocyte biochemistry and function, our understanding of the regulatory system controlling megakaryocytopoiesis is becoming more clear. The cloning and the expression of thrombopoietin, the regulator of megakaryocytopoiesis, has been a major breakthrough. This review will discuss megakaryocyte ontogeny, cell-cell interactions between megakaryocytes and the other stromal cells and the signal transduction in megakaryocytes. PMID- 9728801 TI - Hemoglobin levels correlate with serum soluble CD23 and TNF-Rs concentrations in patients with rheumatoid arthritis. AB - The present study was designed to investigate the possible relationships of hemoglobin concentrations with serum levels of soluble CD23 molecules (sCD23) and soluble tumor necrosis factor receptors I and II (sTNF-RI and sTNF-RII) in rheumatoid arthritis (RA) patients. Fifty-six patients, eight males and 48 females, and 20 age and sex matched healthy volunteers were enrolled in the study. Patients were classified in two groups on the basis of disease activity: group A included 43 patients with active, and group B 13 patients with non-active RA. Serum sCD23 and sTNF-Rs levels were measured using commercially available micro-ELISA kits. It was found that patients of group A had lower hemoglobin concentrations than patients of group B or normal controls, whereas hemoglobin levels in patients of group B did not differ statistically from the controls. Patients of group A had also significantly higher serum sCD23, sTNF-RI and sTNF RII concentrations than patients of group B or control subjects. Serum levels of all three cytokines did not differ statistically between patients of group B and normal controls. In the entire group of subjects studied, hemoglobin concentrations correlated inversely with the levels of serum sCD23, sTNF-RI and sTNF-RII, and also with the values of the erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) reflecting disease activity. We concluded that anemia and elevated concentrations of sCD23, sTNF-RI and sTNF-RII in RA patients are two biological expressions of the same underlying inflammatory process, although a causal relationship between themselves cannot be excluded and needs further investigation. PMID- 9728802 TI - The IL-6 gene expression by leukemic cells from acute lymphoblastic leukemia common and T type and modulation of IL-6 production by TNF. AB - The tumour necrosis factor alpha (TNF-alpha) and interleukin-6 (IL-6) mRNA accumulation and the release of the cytokines TNF-alpha and IL-6 was determined in leukemic cells isolated from bone marrow biopsy from patients with acute lymphoblastic leukemia: ALL-common type (cALL), 11 patients; ALL-T type, nine patients. The non-leukemic bone marrow cells (BMMC) and peripheral blood mononuclear cells (PBMC) from healthy donors were used as a control. The mRNA was assessed by fluorescent in situ hybridization in cell suspension and analyzed with flow cytometry. The accumulation of cytokine mRNA was higher in cALL cells as compared to ALL-T and PBMC (control) and was comparable to cytokines mRNA accumulation in BMMC. The production of IL-6 by leukemic cells from both types of leukemia was significantly lower than in BMMC. The bioactive TNF was not detected in either of the leukemia groups studied. TNF-alpha protein was produced by ALL-T cells and BMMC but not by cALL type of leukemic cells. The synthesis of IL-6 was significantly enhanced by TNF-alpha in BMMC and ALL-T while the presence of TNF alpha had no effect on IL-6 synthesis in the culture of cALL leukemic cells. It was concluded that despite IL-6 and TNF-alpha mRNA contents, leukemic cells representing early stage of B-cell development (CD10+) showed disregulation of production of these cytokines. PMID- 9728803 TI - Effects of daunorubicin on cell growth, cell cycle and induction of apoptosis in HL-60 cells. AB - Administration of daunorubicin (DNR) at over 3.5 x 10(-8)M has been reported to block cells at G2 phase, but the precise mechanism of cell death induced by DNR is not well known. In this study effects of DNR at various concentrations on cell growth, cell cycle and induction of apoptosis in human leukemia cell line HL-60 cells were investigated. Administration of DNR at a high concentration (3.0 x 10( 8)M) inhibited cell growth to 3% as compared with untreated cells, blocked the cell cycle at G2 phase and induced cell-cycle non-specific apoptosis. Administration of DNR at a lower concentration (1.0 x 10(-8)M) inhibited cell growth to 77%, induced cell-cycle nonspecific apoptosis but did not produce G2 arrest. A longer duration of exposure to DNR was required to induce apoptosis by the lower concentration of DNR than by the higher concentration. These findings indicate that the effect of DNR on cell growth is caused by both G2 arrest and induction of apoptosis and that DNR induced apoptosis without G2 arrest. Moreover, continuous administration of a low concentration of DNR which has been considered to have no anticancer effect might be useful for treatment of leukemia and related diseases. PMID- 9728804 TI - The relationship of bone marrow histology with the molecular pattern in chronic myeloid leukemia. AB - The chromosomal abnormality in chronic myeloid leukemia (CML) results from a reciprocal translocation between chromosomes 9 and 22 transferring the c-abl proto-oncogene from chromosome 9 to the restricted breakpoint region on chromosome 22 M (bcr). In this study the breakpoint was determined within the M bcr in 35 CML patients in the chronic phase, by Southern blotting analysis, and it was then correlated with bone marrow Granulocytic-Megakaryocytic (GRAN-MEG) and Granulocytic (GRAN) histological subgroups, as well as with the clinical findings and laboratory parameters. In the 35 patients analyzed, 46% were grouped as 5' and 54% as 3'. There was an increase in bone marrow basophils in 5' breakpoint patients compared to 3' breakpoint (p = 0.042) but the M-bcr breakpoint site did not differ significantly in the subgroup GRAN or GRAN-MEG (p = 0. 12). In conclusion, the patient population had a higher frequency of M-bcr breakpoint in zone 4 and 3' position; there was no correlation between 5' and 3' positions and clinical or haematological features, except a significant increase in bone marrow basophil cells in 5' breakpoint patients compared to 3' breakpoint. Although a higher frequency of the 3' breakpoint was found in patients with a low number of megakaryocytes compared to the cases with a granulocytic-megakaryocytic proliferation, this difference was not statistically significant. PMID- 9728805 TI - Serum erythropoietin levels in patients with leukemia on cytostatic treatment. AB - Anemia is the major stimulus for erythropoietin (Epo) secretion. Various studies have reported increase of Epo following chemotherapy. The mechanism of this phenomenon is not yet clarified. In this study, the serum Epo levels have been evaluated before, during (7 and 14 days), and after (day 25) chemotherapy in patients with acute myeloblastic leukemia (n = 13) and lymphoblastic leukemia (n = 4). As a control group, 12 healthy age-matched subjects were evaluated. Epo levels were high in untreated leukemia patients compared to controls and continued to increase following chemotherapy. There was no significant difference in post-treatment values of Epo as compared with pre-treatment levels. In patients with pre-treatment values of Hb < or = 9 g/dl, Epo levels were inversely correlated with Hb (r = 0.552; p < 0.05). This correlation disappeared during and following treatment. There was no correlation between Epo levels and hematological or biochemical parameters. Therefore, elevated levels of Epo regardless of anemia may be due to a response to tissue hypoxia or increased synthesis of Epo in liver or bone marrow. PMID- 9728806 TI - Characterization of anti-erythrocyte autoantibodies in non-Hodgkin's lymphoma patients in Brazil. AB - The existence of an association between autoimmune phenomena and lymphoproliferative neoplasms is well known. In Campinas at the University Hospital, seventy-seven adult patients with non-Hodgkin's lymphoma (NHL) were studied at diagnosis. The histological subgroup of NHL was performed using Kiel criteria and all patients were characterized by clinical and laboratory examinations according to the Ann Arbor staging. The results of the immunohaematological evaluation of our patients with NHL showed that: 28% presented erythrocyte autoantibodies (auto anti-I or auto-IgG without specificity) but only one developed haemolytic anaemia. There was a weak correlation between low-grade lymphoma and erythrocyte autoantibodies. PMID- 9728807 TI - Severe aplastic anaemia relapsing during a pregnancy; spontaneous remission following termination. AB - The case of a 29-year-old woman with aplastic anaemia in remission who relapsed during pregnancy is reported here. Following successful Caesarean delivery, spontaneous remission was obtained and the patient remains well thereafter. Pathogenetic and therapeutic aspects of this rare complication of pregnancy are discussed. PMID- 9728808 TI - Systemic lupus erythematosus presenting as myelofibrosis. AB - Myelofibrosis is not frequent in systemic lupus erythematosus (SLE). A review in the literature reveals that the co-incidence is rather rare since there are only a few papers reporting this combination. The female patient described hereby, presented with thrombocytopenia; following investigation, the diagnosis of SLE was established and bone marrow examination revealed an increase of marrow reticulin. Treatment with steroids reversed both thrombocytopenia and bone marrow fibrosis. PMID- 9728809 TI - Sweet's syndrome associated with Hodgkin's disease. PMID- 9728810 TI - Relationships of peri-partum, plasma concentrations of progesterone, oestrogens and 13,14-dihydro-15-ketoprostaglandin F2alpha in heifers and of anatomical measurements of dam and calf with difficulty of calving in early-bred Hereford x Friesian heifers. AB - Plasma concentrations of progesterone, oestradiol-17beta, oestrone, oestrone sulphate and PGFM have been measured daily during the first peri-partum period of 45 Hereford x Friesian heifers bred at 11 months of age. Anatomical measurements of dam and calf were also recorded. Twelve of the calvings were scored easy, 33 difficult. Each of five models (fitted by linear logistic regression) relating difficulty of calving to the hormonal and anatomical measurements, predicts with at least 94% accuracy the calving score (easy or difficult) among the calvings. The models predict that increases of progesterone concentration on the day before calving, of oestrone sulphate concentration on the day after calving and of heifer heart girth decrease the odds of difficult calving, whereas increases of heifer body length and of calf head circumference increase the odds of difficult calving. PMID- 9728811 TI - Development of tumors from bovine primordial germ cells transplanted to athymic mice. AB - Intact genital ridges containing primordial germ cells (PGC) and isolated PGC from murine and bovine embryos were examined for in vivo growth and differentiation after transplantation under the kidney capsule of athymic mice. Genital ridges were collected from day 11.5 and 12.5 murine and day 34 and 37 bovine embryos. Murine genital ridges and isolated PGC collected at 11.5 days post-coitus (dpc) and isolated murine PGC collected at 12.5 dpc developed into tumors. Day 34 and 37 bovine genital ridges, but not isolated PGC developed into tumors. The bovine origin of the tumors was confirmed by an analysis of the bovine DNA sequences. Tumors from both species consisted primarily of mesoderm derived cell types, including connective tissue, cartilage, smooth muscle, fibroblasts, osteoblasts and bone matrix. No detectable ectodermal derivatives were observed in tumors of either species. Undifferentiated stem cells were not detected in the tumors, suggesting that the tumors were benign teratomas. Results of this study demonstrate the pluripotency of bovine PGC by experimental induction of teratomas after xenotransplantation under the kidney capsule of athymic mice. Stimulation of PGC survival and proliferation in an ectopic graft could be useful toward the isolation of pluripotent embryo-derived stem cells. PMID- 9728812 TI - Plasma beta-endorphin around parturition and its relationship to cortisol level and resumption of pituitary and ovarian functions in dairy cows. AB - The objectives were to evaluate the concentrations of beta-endorphin in peripheral circulation around parturition and to investigate their relationship to the concentrations of cortisol and postpartum resumption of pituitary and ovarian functions in dairy cows. Subjects were 21 Holstein-Friesian cows in late pregnancy. Blood samples were collected from these animals from day 270 in pregnancy until the first ovulation after calving. Average immunoreactive (IR) beta-endorphin concentrations in cows with dystocia (n = 8) in periparturient period (from day 270 of pregnancy until 24 h after calving) were slightly higher than those in cows with normal calving (n = 13) in the same period although the difference was not significant. During the periparturient period, the peak level of IR beta-endorphin was seen at the time of rupture in cows with normal calving and it was observed at the time of parturition in cows with dystocia. The trend of IR beta-endorphin secretion appeared to be concomitant with cortisol secretion in the periparturient period but not in postpartum period (from 24 h after calving until the first ovulation). Average IR beta-endorphin concentrations in cows with dystocia in the postpartum period were not significantly different from those in cows with normal calving. There was no significant association between average IR beta-endorphin concentrations in early postpartum period (from 24 h until 7 days after calving) and the responsiveness of luteinising hormone (LH) to exogenous GnRH administered on day 7 postpartum. However, a negative correlation (r = -0.593; n = 21; P = 0.004) was observed between average IR beta-endorphin concentrations and LH concentrations during the period from day 1 until the first ovulation in the 21 cows. In addition, a positive correlation (r = 0.498; n = 21; P = 0.020) was seen between the beta-endorphin concentrations and time to the first ovulation after calving. The results from this study suggest that beta endorphin may release into peripheral blood differently from cortisol and that it may be involved in regulating LH secretion and thus the resumption of ovarian cyclicity in postpartum dairy cows. PMID- 9728813 TI - Comparison of two co-culture systems to assess the survival of in vitro produced bovine blastocysts after vitrification. AB - The ability of bovine blastocysts to recover after cryopreservation and thawing procedures is often assessed by evaluating their re-expansion during in vitro co culture. However, the influence of factors such as feeder cell type and gas atmosphere on blastocyst survival and evolution have never been considered. This study therefore compared two cell co-culture systems and two different gas atmospheres to assess survival of in vitro produced bovine blastocysts after vitrification. Day-7 blastocysts (n = 181) were vitrified in a mixture of 25% glycerol/25% ethylene glycol. After warming and dilution, they were co-cultured either on Buffalo rat liver cells (BRL CC cell line) or on granulosa cells (GR CC primary culture) in TCM 199 supplemented with 10% FCS and under an atmosphere of 5% or 20% O2. Surviving and hatching rates were recorded at 24 h intervals for 3 days. After 72 h of culture, surviving blastocysts were treated for differential counting of inner cell mass (ICM) and trophectoderm cells. Blastocyst survival rates were higher when BRL and granulosa co-culture were performed under 20% oxygen as compared to 5% oxygen (20% O2: 62% vs. 5% O2: 25%, P < 0.0001). However, the quality of blastocysts surviving in the granulosa co-culture condition was lower under 20% O2 than under 5% O2 as indicated by lower total and trophectoderm cell numbers (respectively 79 +/- 6 and 56 +/- 6 at 20% O2 vs. 100 +/- 10 and 74 +/- 10 at 5% O2, P < 0.05), by an altered ICM/trophectoderm ratio (20% O2: 28% vs. 5% O2: 23%, P < 0.05), by a higher total nuclear fragmentation (20% O2: 3.7% vs. 5% O2: 1.5%, P < 0.05) and a trend to decreased hatching (20% O2: 32% vs. 5% O2: 81%, P = 0.07). Whereas, for BRL co-culture, 20% O2 yielded higher quality blastocysts than 5% O2 as evaluated by higher ICM and trophectoderm cell numbers (19 +/- 1 and 71 +/- 5 at 20% O2 vs. 15 +/- 2 and 48 +/- 9 at 5% O2, respectively, P < 0.05), by lower nuclear fragmentation in the ICM (20% O2: 2.2% vs. 5% O2: 6.7%, P < 0.05). In conclusion, co-culture conditions may influence blastocysts survival and quality after cryopreservation. In our conditions, co-culture with BRL cells under 20% O2 seems to be the best combination to evaluate blastocyst survival and quality after vitrification. PMID- 9728814 TI - Differential effects of kinase inhibitor and electrical stimulus on activation and histone H1 kinase activity in pig oocytes. AB - The 6-DMAP (6-dimethylaminopurine) has been reported to accelerate and enhance the formation of pronuclei (activation) in non-aged MII mouse and bovine eggs. In this study, effects of chemical activation (CA) were evaluated using ethanol + 6 DMAP and electrical activation (EA) on pig oocytes matured for 36 h (newly matured) and 48 h. After CA, the first pronuclei developed within 2 h, with maximal pronuclear formation at 8 h (> 90%) for oocytes matured for either 36 or 48 h. In addition, chemically activated oocytes did not extrude the second polar body. After EA treatment, pronuclear formation began at 4-6 h and was significantly lower at 8 h in newly matured than aged oocytes (34% vs. 85%). Cleavage rate at 24 h after treatment was lower for oocytes treated by CA (< 50%) than for EA (> 70%) and after 3 days of in vitro culture, fewer oocytes reached the four-cell stage in CA than in EA groups. No chemically activated oocytes developed beyond the four-cell stage; in contrast, EA resulted in development beyond this stage. Both CA and EA resulted in a drop of H1 kinase activity, but EA induced a more rapid reduction. With EA, H1 kinase activity rapidly declined but tended to increase again at 8 h in oocytes matured for 36 h, which was different from all other groups in which the kinase activity remained low at 8 h post-activation. We conclude that different mechanisms are involved in pronuclear formation with CA and EA and that 6-DMAP-activated oocytes may not be suitable for the induction of normal parthenogenetic development in pigs. It is also suggested that the ability of pig oocytes to become fully activated may be related to the modification of H1 kinase activity during the aging process. These modifications to H1 kinase may be necessary for parthenogenetic activation of in vitro matured oocytes and also for the completion of meiosis and cleavage. PMID- 9728815 TI - Effect of vulvomucosal injection of PGF2alpha at insemination on subsequent fertility and litter size in pigs under field conditions. AB - The aim of this study was to determine the effects of injecting 5 mg of PGF2alpha into the vulvar mucosa on the reproductive performance of sows maintained under field conditions. At an intensively managed piggery in northwest Spain, two experimental groups were formed randomly and observed throughout the year. The first group comprised sows receiving simultaneously, with every insemination, 5 mg of PGF2alpha injected into the vulvar lips. Group 2 sows received 1 ml of saline solution injected into the vulvar lips at insemination and served as the controls. The farrowing rates for each group were 78.46 and 54.39, while the litter sizes were 10.72 +/- 0.27 and 9.14 +/- 0.47 during the low fertility season (July-September). During the rest of the year (October-June), the farrowing rates were 83.91 and 80.93, while the litter sizes were 11.16 +/- 0.15 and 9.99 +/- 0.15. We conclude that injection of 5 mg of PGF2alpha into the vulvar lips at insemination is an effective method of compensating for the low fertility together with the decreased litter size of the summer months. PMID- 9728816 TI - The effects of post-weaning progestagen treatment (Regumate) of early-weaned primiparous sows on subsequent reproductive performance. AB - This study investigated the effects of feeding the orally active progestagen, altrenogest (Regumate) post-weaning on the subsequent reproductive performance of early weaned sows. Ninety (90) Large White/Landrace first parity sows were randomly assigned to three treatments. Treatment 1 (EW) and treatment 3 (CW) sows were weaned on day 12 and day 24 post-partum, respectively while treatment 2 sows (EW-R) were weaned on day 12 post-partum and received an individual daily dose of 20 mg of Regumate on days 13 to 24 post-partum inclusive. Each sow was mated naturally at least twice at the first post-weaning or post-treatment oestrus and slaughtered on days 25-28 of pregnancy to determine the number of corpora lutea and embryos. Regumate-to-oestrus and weaning-to-oestrus intervals were similar for EW-R and CW sows (6.2 vs. 5.6 days). However, both intervals were significantly shorter (P < 0.01) than the weaning-to-oestrus interval of EW sows (7.3 days). An excellent synchronization of oestrus was achieved with Regumate treatment with 97% of treated sows in oestrus within 7 days of Regumate withdrawal compared with 64% for EW sows (P < 0.01) and 87% for CW sows (P > 0.05). Treatment with Regumate resulted in a significant increase in ovulation rate (16.9 vs. 15.4 and 14.9 for treatments EW-R, EW and CW, respectively; P < 0.05) and a non-significant increase in early embryonic survival (77% vs. 68% vs. 68% for treatments EW-R, EW and CW, respectively; P > 0.05). These results indicate that Regumate feeding is a potential management tool to alleviate the diminished reproductive performance associated with early weaning regimes since it leads to successful control of oestrus, higher ovulation and embryo survival rates and thus a greater potential litter size. PMID- 9728817 TI - Steroid metabolism in granulosa and theca interna cells from preovulatory follicles of domestic hen (Gallus domesticus). AB - The capability of granulosa and theca interna cells, from preovulatory follicles of the domestic hen, to metabolize steroid precursors was evaluated. Granulosa and theca interna cells were isolated from ovarian preovulatory follicles at three different developmental stages: F1, F3 and F5. Tritiated pregnenolone (P5), progesterone (P4), dehydroepiandrosterone (DHEA), androstenedione (A4) and testosterone (T) were employed as precursors and their metabolic products were evaluated. The major metabolite of P5 by granulosa cells was P4, but we also observed low amounts of 5beta-pregnandione. DHEA metabolism by granulosa cells yielded mainly A4, and minute quantities of 5beta-androstan-3,17-dione (5beta dione) were detected. The only significant metabolite obtained in granulosa cells from A4 was 5beta-dione, whereas T was only transformed into A4. On the other hand, P5 metabolism by theca interna cells yielded A4 as the main product, also P4, 17alpha-OHP4, 17alpha-OHP5, 5beta-pregnandione, and DHEA, were found. When DHEA was the precursor A4 was produced in higher amounts than 5beta-dione. A4 was mainly transformed into 5beta-dione. In similar conditions, T was transformed into A4. These results show that granulosa cells have enzymatic activities of 3beta-hydroxysteroid dehydrogenase/5-4 isomerase (3beta-HSD from P5 and DHEA), 17beta-hydroxysteroid dehydrogenase (17beta-HSD from T) and 5beta-reductase (from P5, DHEA and A4). Whereas theca interna cells have enzymatic activities of cytochrome P450c17 (from P5 and P4), 3beta-HSD (from P5 and DHEA), 17beta-HSD (from T) and 5beta-reductase (from P4, DHEA and A4). These data support the concept that theca interna cells have the ability to synthesize androgens from progestins produced in granulosa cells. In addition, since theca interna cells did not show the capacity to aromatize androgens suggests that interaction between theca interna and theca externa cells occurs in vivo, thus confirming the three cell model for estrogen production. Furthermore, the fact that other metabolites were produced both in granulosa and theca interna cells, but in a different extent, suggests that complex mechanisms are participating in the regulation of steroid synthesis in avian ovary follicles. PMID- 9728818 TI - The mechanism of alpha2-adrenergic inhibition of sympathetic ganglionic transmission. AB - Alpha2-adrenergic agonists produce analgesia and reduce hemodynamic stress through central and peripheral mechanisms, but the effect of adrenergic agonists on pre- and postganglionic sites has not yet been clarified. In this study, we examined the effects of dexmedetomidine (DMT), an alpha2-agonist, on neural conduction and neurotransmitter release in sympathetic ganglia. The stellate ganglia from 48 mongrel dogs were isolated, desheathed, and superfused with Krebs' solution. Compound action potentials were evoked, and chromatography was used to detect acetylcholine released by preganglionic stimulation in the presence or absence of DMT. To further elucidate the mechanism of alpha2 effects, DMT was applied in combination with the alpha2-antagonist atipamezole (AT) or the imidazoline antagonist idazoxan (ID). In other experiments, DMT was applied in the presence of exogenous nicotinic stimulation with 1,1-dimethyl-4 phenylpiperazinium iodide or muscarinic stimulation with (+)cis-dioxolane. DMT dose-dependently inhibited synaptic transmission with a 50% effective dose of 71.6 (26.0-174.3) microM. Neurotransmitter release was reduced 25% by 70 microM DMT during low-frequency (0.4 Hz) stimulation, but this effect was abolished at higher frequency (5 Hz) stimulation. AT but not ID blocked the inhibitory action of DMT. DMT inhibited the excitatory postsynaptic response to exogenous muscarinic stimulation but not nicotinic stimulation. These results indicate that alpha2-receptor activation depresses ganglionic transmission through postsynaptic inhibition of muscarinic stimulation, although reduction of neurotransmitter release through a presynaptic autofeedback mechanism is also involved. IMPLICATIONS: This article provides novel insights into the mechanism of drug action of alpha2-receptor agonists in the sympathetic ganglia of dogs by directly measuring the relative contribution of pre- and postganglionic receptors. Our study indicates that the central sympatholytic effects of alpha2-adrenoceptor stimulation are augmented by peripheral inhibition of ganglionic transmission. PMID- 9728819 TI - Disparity of isoflurane effects on left and right ventricular afterload and hydraulic power generation in swine. AB - The interaction between myocardial and vascular effects of anesthetics has a potential impact on how these drugs influence performance of the heart. Most studies have focused on volatile anesthetic effects on the left ventricle (LV) and systemic circulation. Whether the right ventricle (RV) and pulmonary circulation respond in a similar fashion, however, is unclear. In the present study, we therefore examined the dose-related effects of isoflurane on LV and RV contractility and total afterload and related changes to simultaneous effects on the hydraulic power generated by each chamber. Two groups of swine were studied: one received no additional treatment before isoflurane (ISO, n = 6), and the other received hexamethonium, atropine, and propranolol to produce autonomic blockade before isoflurane administration (ISO+AB, n = 4). For each experiment, measurements were made of RV and LV regional segment lengths and pressures, along with proximal aortic and pulmonary arterial (PA) blood flow and pressure during the administration of 0, 0.5, 1.0, and 1.5 minimum alveolar anesthetic concentration (MAC) isoflurane. Contractility was assessed by calculating the regional preload recruitable stroke work slope (PRSW). Afterload was characterized in both nonpulsatile and pulsatile terms by calculating aortic input impedance magnitude (Z). From these data, total arterial resistance (R), characteristic impedance (ZC), and vascular compliance (C) were determined with reference to a three-element Windkessel model of the circulation. Additionally, steady-state (WSS), oscillatory (WOS), and total (WT) hydraulic power output of each ventricle was calculated. In the ISO group, isoflurane produced a nearly threefold greater decrease of peak systolic pressure in the LV than in the RV, yet the dose-related decrease of regional PRSW was virtually the same in both chambers. In the aorta, isoflurane produced a maximal 25% reduction in R at 1.0 MAC and doubled C without a significant change in ZC. Alternatively, PA R was increased from baseline at 1.0 and 1.5 MAC, whereas ZC was increased from all other values at 1.5 MAC. PA C was not altered by isoflurane. In ISO+AB pigs, PA ZC at baseline was higher than that evident in ISO animals but was not altered by isoflurane. In contrast, baseline aortic R was lower in ISO+AB pigs but was still modestly reduced by 1.0 MAC isoflurane. In ISO animals, WT and WSS from both ventricles demonstrated dose-related decreases, but the reductions in LV WTand WSS were greater than those for the RV at all doses. Accordingly, the power requirement per unit flow decreased for the LV but remained constant for the RV. WOS for both ventricles was also reduced by isoflurane. However, the LV WOS to WT ratio increased, which indicates that more power was lost to the system by pulsation. In contrast, reductions in RV WT and WOS were nearly parallel at all isoflurane doses, and the WOS to WT ratio was unchanged. In the ISO+AB group, isoflurane-induced alterations in LV and RV power characteristics were similar to those in the ISO group. These data indicate that, despite similar effects on biventricular contractility, isoflurane exerts qualitatively different effects on RV and LV afterload, in part via alteration in autonomic nervous activity, that influence the distribution of power output between steady-state and pulsatile components. IMPLICATIONS: In this study, we examined the effects of isoflurane on cardiac performance in swine and found that, although the drug depresses contraction of both the left and right ventricles similarly, it has different effects on forces that oppose the ejection of blood. These findings demonstrate that the two interdependent pumps that comprise the heart can be influenced differently by anesthetic drugs. PMID- 9728820 TI - The effects of desflurane on cardiac function as measured by conductance volumetry in swine. AB - The purpose of the investigation was to assess the effects of desflurane (DES) on left ventricular heart function during basal barbiturate anesthesia in a closed pericardium, closed-chest acute swine model. The study was performed in 11 normoventilated adult pigs. Hemodynamic measurements were obtained using arterial, central venous, and pulmonary artery catheters, as well as a conductance volumetry and tip manometry catheter placed in the left ventricle. Hemodynamic measurements were recorded during basal pentobarbital anesthesia and with the addition of 1%, 2%, 4%, and 6% DES. DES dose-dependently decreased mean arterial pressure, systemic vascular resistance, left ventricular end-systolic pressure, dP/dtMAX and dP/dtMIN. At doses >1%, decreases in CO, stroke volume, ejection fraction, end-systolic elastance, preload recruitable stroke work, preload adjusted maximal power, and peak filling rate were observed. Heart rate decreased at 4% and 6% DES. Isovolumetric relaxation time increased only at 6% DES. We conclude that smaller doses of DES have a significant cardiodepressive effect in the setting of barbiturate infusion, as measured by conductance volumetry. IMPLICATIONS: Desflurane, in very small doses, depressed cardiac function during pentobarbital anesthesia with ketamine and benzodiazepine premedication in swine, as assessed by conductance volumetry and left ventricular pressure and volume relationship analysis. These results suggest that desflurane, in combination with certain anesthetics, can be cardiodepressive even in very small doses. PMID- 9728821 TI - Elective splenectomy in an anemic Jehovah's Witness patient with cirrhosis. PMID- 9728822 TI - The use of a video to convey preanesthetic information to patients undergoing ambulatory surgery. AB - In a prospective, randomized, single-blind trial, we assessed the effectiveness of a preoperative video as a source of additional patient information before ambulatory surgery. One hundred twenty-seven patients were allocated to either treatment (video) or control (nonvideo) groups. Of the 127, 17 (13%) patients correctly answered all process, risk, and misconception statements using a questionnaire. Overall, the video group was 2-16 times more likely to recall all knowledge questions correctly than the nonvideo group after adjusting for previous general anesthesia experience, state (how one feels at the moment), and trait (how one generally feels) anxiety levels (relative risk 6.36, 95% confidence interval 2.01-15.82). The predictors of correct risk knowledge were those who had a video intervention (relative risk 7.12, 95% confidence interval 3.70 to 10.07) and low trait anxiety scores (relative risk 5.88, 95% confidence interval 1.69 to 25.00). A video could be an important additional component of the preoperative interview, but anesthesiologists will still need to provide patient-specific information. IMPLICATIONS: This study randomly allocated adults to see a video about anesthesia before scheduled ambulatory surgery. The video group had better recall of information. The video was a useful adjunct to routine preoperative consultations. PMID- 9728823 TI - Predictive factors of outcome in severely traumatized children. AB - To identify risk factors associated with death in traumatized children, we prospectively studied 507 consecutive patients (7+/-4 yr) admitted to a level I pediatric trauma center over a 3-yr period. Pediatric Trauma Score (PTS), Glasgow Coma Scale (GCS) score, and Injury Severity Score (ISS) were calculated. Age, injury mechanism, injury pattern, and initial critical care were recorded. Univariate and multivariate analyses were performed for potential risk factors associated with mortality. Receiver operating characteristic curves were used to determine threshold values of variables identified by univariate analysis. Most children suffered from blunt trauma (99.6%), and head trauma was noted in 85%. Median values (range) of GCS scores, PTS, and ISS were 10 (3-15), 7 (-4 to 12), and 16 (3-75), respectively. The mortality rate was 12%. Using multivariate analysis, death was significantly associated with an ISS > or = 25 (odds ratio [OR] 22.2, 95% confidence interval 2.8-174.9), GCS score < or = 7 (OR 4.77, 1.8 12.7), emergency blood transfusion > or = 20 mL/kg (OR 4.3, 2.1-9.1), and PTS < or = 4 (OR 3.7, 1.4-9.7). An ISS > or = 25, GCS score < or = 7, immediate blood transfusion > or = 20 mL/kg, and PTS < or = 4 were significant and independent risk factors of death in an homogenous population of severely injured children. The probability of traumatic death was therefore 0 (95% confidence interval 0 0.0135) in children with no one of these threshold values in the four predictive factors and 0.63 (95% confidence interval 0.47-0.76) in those children with all the threshold values. IMPLICATIONS: Methods used for evaluating outcome of trauma patients have essentially been derived from adult series, and attempts to apply them to children have usually been inaccurate. Univariate and multivariate analyses were performed to identify risk factors associated with death in severely traumatized children, and Receiver operating characteristic curves were used to determine threshold values. PMID- 9728824 TI - Effects of cardiopulmonary bypass and deep hypothermic circulatory arrest on the thyroid axis during and after repair of congenital heart defects: preservation by deep hypothermia? AB - Thyroid function is altered by cardiopulmonary bypass (CPB) in children. To better understand the cause of altered thyroid hormone levels, we compared the effects on the pituitary-thyroid axis of CPB in 23 children undergoing elective repair of congenital heart defects. Twelve patients underwent CPB with moderate hypothermia without a period of deep hypothermic circulatory arrest (DHCA), and eleven underwent CPB with DHCA. Nine blood samples were collected from each patient before, during, and after CPB. Free T3 (FT3), free T4 (FT4), total T3 (TT3), total T4 (TT4), thyrotropin (TSH), and albumin were measured; concentrations of each decreased significantly with the onset of CPB (P < 0.05). There was a greater decline in hormone than in albumin concentrations, which suggests that factors in addition to hemodilution were present (P < 0.05). TSH concentrations in the DHCA group began to increase during cooling, exceeding baseline values after rewarming and after separation from CPB. Patients undergoing CPB without DHCA had persistently low TSH concentrations (P < 0.05). By Postoperative Days 1 and 2, TSH concentrations in both groups were similar and significantly lower than baseline values (P < 0.001). FT3, FT4, TT3, TT4, and albumin all increased during CPB after an initial decrease. Of these, only albumin and FT4 recovered to their baseline values after the initial decrease. Nevertheless, by Postoperative Day 1, both groups demonstrated the "sick" euthyroid syndrome and could not be distinguished from one another. This study demonstrates greater pituitary release of TSH in children undergoing repair of congenital heart defects with DHCA compared with CPB alone, the cause of which could not be determined in this study. However, despite the increase in TSH in the DHCA group, the thyroid hormone concentrations failed to increase appropriately. IMPLICATIONS: Early after deep hypothermia circulatory arrest, thyrotopin concentrations increase appropriately, responding to decreased concentrations of T3; however, all children undergoing cardiopulmonary bypass eventually develop a "sick" euthyroid syndrome by Postoperative Day 1. Whether this difference represents better protection of neuroendocrine function by deep hypothermic circulatory arrest (relative to cardiopulmonary bypass alone) remains speculative. PMID- 9728825 TI - Simulation of an epidural test dose with intravenous isoproterenol in sevoflurane and halothane-anesthetized children. AB - Isoproterenol has been suggested as an alternative marker for epidural test dosing in children receiving halothane anesthesia. The purpose of this prospective, randomized, double-blind study was to determine the chronotropic response to IV isoproterenol in sevoflurane-anesthetized children. Thirty-six ASA physical status I children (0.5-8 yr) were anesthetized with either halothane or sevoflurane at 1 minimum alveolar anesthetic concentration adjusted for age in 70% nitrous oxide. Patients received incremental IV injections of isoproterenol until their heart rate increased > or = 20 bpm above baseline. The minimal effective dose of isoproterenol required to produce an increase of > or = 20 bpm was 55 ng/kg (42-72 ng/kg; 95% confidence interval) in sevoflurane-anesthetized children and 32 ng/kg (26-38 ng/kg; 95% confidence interval) in halothane anesthetized children (P < 0.05). This dose-response study suggests that sevoflurane antagonizes beta-adrenergic-mediated chronotropic responses to isoproterenol more than halothane. These observations also suggest that larger doses of isoproterenol will be necessary for epidural test dosing in children receiving sevoflurane rather than halothane anesthesia. IMPLICATIONS: Isoproterenol has been suggested as an alternative marker for epidural test dosing in children receiving halothane anesthesia. This isoproterenol dose response study indicates that larger doses of isoproterenol will be necessary for epidural test dosing in children undergoing sevoflurane rather than halothane anesthesia. PMID- 9728826 TI - Delleman syndrome: anesthetic implications. AB - An infant with oculocerebrocutaneous (Delleman) syndrome (1), only 26 cases of which have been reported (2), presented with focal alopecia of scalp, periorbital skin appendages, hypertrophy of the skin (Fig. 1A), left-sided orbital cyst, lid coloboma, cleft palate (Fig. 1B), neonatal seizures, cerebral hemiatrophy, multiple intracranial cystic spaces, and enlarged lateral ventricles. The anomalies often require multiple anesthetics for examination of the eye, drainage of the orbital cyst, repair of lid coloboma, enucleation of the eye, excision of skin tags, and repair of cleft palate. Although this infant's perioperative course was uneventful, he had significant preoperative problems, such as neonatal seizures and an episode of aspiration pneumonia. Because the Delleman syndrome is rare, this case is presented to illustrate possible anesthetic implications of the disease. PMID- 9728828 TI - Baclofen withdrawal presenting as multiorgan system failure. PMID- 9728827 TI - Percutaneous dilational tracheostomy: the Ciaglia method versus the Portex [correction of Rapitrach] method. AB - In a prospective study, we performed percutaneous dilational tracheostomy (PDT) using either the Ciaglia method (gradual dilation) or the Portex [corrected] method (single dilation) in 80 patients. We encountered difficulty in dilating the tracheal stoma of three (7.5%) patients in the Ciaglia group because of tight pretracheal fascia. It was difficult to insert the tracheostomy tube in four (10%) patients in the Portex [corrected] group even after appropriate tracheal dilation. However, the tracheal cannulation was successfully completed in all patients in subsequent attempts. The mean time for completion of the procedures, from skin incision to insertion of the tracheostomy tube, was 14+/-5.5 min with the Ciaglia method and 6.5+/-3.5 min with the Portex [corrected] method. PDT with either method has not been associated with clinically significant hemorrhage, infection at the stoma site, or cosmetic deformity. In a follow-up period of 9 mo, none of our decannulated patients presented with clinical tracheal stenosis. Our results indicate that PDT with both methods is as safe and easy to organize and perform as a bedside procedure, obviating the need to transport critically ill patients from the critical care unit. IMPLICATIONS: The tracheas of 80 patients were cannulated through an artificial opening using either the Ciaglia (gradual dilation) or the Portex [corrected] (single dilation) method. Both techniques were successful with no significant complications. After 9 mo of closure of this opening, none of the survivors had significant scarring or narrowing of the trachea. PMID- 9728829 TI - Propofol and hyperventilation for the treatment of increased intracranial pressure in rabbits. AB - Using a rabbit model of intracranial hypertension, we studied (a) the additive effect of propofol and hyperventilation (HV) on increased intracranial pressure (ICP), (b) the ICP-lowering effect of additive therapy (i.e., propofol plus HV versus HV plus propofol), and (c) whether combined therapy induced cerebral ischemia. Twenty-three New Zealand White rabbits were studied, of which seven were control animals. The animals were anesthetized with midazolam and fentanyl. An extradural balloon was used to increase ICP to 26+/-2 mm Hg. Elevation of ICP resulted in a >50% reduction in ipsilateral somatosensory evoked potential (SEP) amplitude. The rabbits were then randomized to be treated with propofol followed by HV (Group 1, n = 8) or HV then propofol (Group 2, n = 8). With HV, PaCO2 was reduced from 41+/-2 mm Hg to 27+/-3 mm Hg. In Group 1 rabbits, the ICP decreased with treatment (from 26+/-2 to 12+/-2 mm Hg). The reduction in ICP was significantly greater (P = 0.008) than that in Group 2 rabbits (from 26+/-2 to 16+/-5 mm Hg). In both groups, propofol and HV had an additive effect in reducing ICP. Further, when propofol was used as the initial treatment, there was a significant increase in SEP amplitude that was not apparent in Group 2 rabbits. In conclusion, we found propofol and HV to be additive in the treatment of increased ICP. In animals treated with propofol followed by HV, there was a greater decrease in ICP than in rabbits treated initially with HV and then propofol. IMPLICATIONS: We have demonstrated that propofol has a greater effect on increased intracranial pressure than hyperventilation when used as the initial treatment and that the two treatments are additive. Should control of intracranial pressure be required after the administration of propofol, then hyperventilation may be added to the treatment. PMID- 9728830 TI - The effect of remifentanil on cerebral blood flow velocity. AB - In the present study, we investigated the effect of remifentanil on cerebral blood flow velocity (CBFV). We investigated 20 patients (ASA physical status III) scheduled for elective coronary artery bypass graft surgery. Anesthesia was induced with remifentanil 5 microg/kg IV (Group 1, n = 10) or 2 microg/kg IV (Group 2, n = 10) and was maintained with 3 microg x kg(-1) x min(-1) IV (Group 1) or 1 microg x kg(-1) x min (-1) IV (Group 2). Pancuronium (0.1 mg/kg IV) was administered for muscle relaxation. Assisted ventilation followed by controlled ventilation via a mask was performed with the PaCO2 kept constant. Mean cerebral blood flow velocity (Vmean) was measured in the middle cerebral artery using a transcranial Doppler sonography system. Mean arterial pressure (MAP) was kept constant by the IV administration of norepinephrine. Measurements were made at baseline and every minute after remifentanil infusion for 10 min. Data were analyzed by using analysis of variance and a post hoc t-test (P < 0.05). Heart rate, MAP, and PaCO2 did not change over time in either group. Vmean did not change in Group 2. In contrast, there was a 31% decrease of Vmean in Group 1 (P < 0.05). The results show that large-dose, but not moderate-dose, remifentanil reduces CBFV unrelated to any changes in systemic hemodynamics in isocapnic cardiac patients. IMPLICATIONS: Transcranial Doppler sonography was used to monitor remifentanil-induced changes in cerebral perfusion. We found that large doses of remifentanil reduced cerebral blood flow velocity despite constant perfusion pressure. This may implicate a central mechanism for cerebral hemodynamic effects of remifentanil. PMID- 9728831 TI - Phenylephrine-induced hypertension does not improve outcome after closed head trauma in rats. AB - Phenylephrine-induced hypertension (increase of 30-35 mm Hg for 15 min) is reported to increase cerebral perfusion pressure and collateral flow to ischemic areas of the brain in a rat model of focal cerebral ischemia. In the present study, we examined whether phenylephrine-induced hypertension of similar magnitude and duration was beneficial in a rat model of closed head trauma (CHT). Forty-eight rats were randomized into four experimental conditions: CHT at time 0 min (yes/no), plus phenylephrine-induced hypertension (increase of 30-35 mm Hg for 15 min) at 65 min (yes/no). CHT was delivered using a weight-drop device (0.5 J). Outcome measures were neurological severity score (NSS) at 1, 4, and 24 h, and brain tissue specific gravity (microgravimetry) and injury volume (2,3,5 triphenyltetrazoium chloride) at 24 h. After CHT, NSS at 24 h (median +/- range) and brain tissue specific gravity (mean +/- SD, injured hemisphere) were 7+/-2 and 1.033+/-0.007 without phenylephrine and 8+/-2 and 1.035+/-0.005 with phenylephrine (P = 0.43), respectively. Tissue injury volume (mean +/- SD) was 335+/-92 mm3 without phenylephrine and 357+/-154 mm3 with phenylephrine (P > 0.62). The results of our study indicate that postinjury treatment with 15 min of phenylephrine-induced hypertension does not attenuate brain edema, reduce tissue injury volume, or improve neurological outcome after CHT in rats. IMPLICATIONS: Phenylephrine-induced hypertension is reported to increase cerebral perfusion pressure and blood flow in a rat model of focal cerebral ischemia. In our study, phenylephrine-induced hypertension did not decrease brain edema or tissue injury volume or improve neurological outcome in a rat model of closed head trauma. PMID- 9728832 TI - The influence of scalp infiltration with bupivacaine on hemodynamics and postoperative pain in adult patients undergoing craniotomy. AB - After craniotomy, hypertension may contribute to intracerebral hemorrhage. We studied whether scalp infiltration with bupivacaine during craniotomy reduces postoperative pain and hypertension. In a double-blind fashion, 36 adult patients (ASA physical status II or III) undergoing elective craniotomy were randomly assigned to receive scalp infiltration with either bupivacaine (0.25%) and epinephrine (1:200,000) or saline/ epinephrine (1:200,000) for skeletal fixation, skin incision, and wound closure. Heart rate (HR) and mean arterial pressure (MAP) were measured after anesthesia induction, after skull-pin insertion, after scalp infiltration, during dural closure, during skin closure, on admission to postanesthesia care unit (PACU), and 1 h after admission. Visual analog pain scores were recorded in the PACU. MAP was significantly greater in the saline group at scalp infiltration. HR was significantly faster in the saline group at dural and skin closure. The bupivacaine group reported significantly less pain than the saline group at PACU admission and 1 h after admission. Pain scores did not correlate with hemodynamic measurements. We conclude that scalp infiltration with 0.25% bupivacaine with 1:200,000 epinephrine blunts certain intraoperative hemodynamic responses and reduces postoperative pain but has no effect on postoperative hemodynamics. IMPLICATIONS: We sought to evaluate whether scalp infiltration with bupivacaine and epinephrine at the beginning and end of craniotomy would afford more intra- and postoperative hemodynamic stability and influence immediate postoperative pain. We found that intraoperative hemodynamics were not influenced greatly; however, craniotomy patients do have significant postoperative pain, which does not seem to have an influence on hemodynamics in the postanesthesia care unit. PMID- 9728833 TI - Antinociceptive effect induced by the combined administration of spinal morphine and systemic buprenorphine. AB - We evaluated the antinociceptive effect of combined spinal administration of morphine and systemic administration of buprenorphine. Experiments were performed on male Wistar rats. Nociception was measured using the tail immersion test. Buprenorphine was injected intraperitoneally (IP) and morphine was injected intrathecally (IT) via a catheter implanted in the subarachnoid space. Interaction of drugs was analyzed using a dose addition model. Both IT (1-5 microg) morphine and IP (50-500 microg/kg) buprenorphine increased the latencies of nociceptive responses in a dose-dependent manner. IT morphine (4 microg) and IP buprenorphine (100 microg/kg) produced 62.9+/-6.3 and 48.8+/-6.6 percent of the maximal possible effect (%MPE), respectively. The combined administration of 2 microg of IT morphine and 50 microg/kg IP buprenorphine produced a %MPE of 97.1+/-3.4. The analysis of drug interaction revealed that IT morphine interacted with IP buprenorphine in a supraadditive manner while producing a potent antinociceptive effect. IMPLICATIONS: The concurrent administration of spinal morphine and systemic buprenorphine produces an antinociceptive effect that is greater than what could have been predicted from individual dose-response curves. This mode of interaction allows maintenance at a significant level of analgesia with reduced doses of opioids, which minimizes the incidence of undesirable side effects. PMID- 9728834 TI - Epidural fentanyl reduces the shivering threshold during epidural lidocaine anesthesia. AB - Epidural local anesthetics and IV opioids both decrease the core temperature that triggers shivering. However, the effect of epidural opioids on shivering thresholds has not been assessed. In this study, we tested the hypothesis that adding epidural fentanyl to epidural lidocaine decreases the shivering threshold compared with epidural lidocaine alone. Fourteen healthy male patients undergoing extracorporeal shockwave lithotripsy under epidural anesthesia were randomly assigned to receive either epidural lidocaine or epidural lidocaine plus epidural fentanyl. Ice-cold lactated Ringer's solution was given IV before epidural blockade, and the core temperature that triggers shivering was established. Then epidural anesthesia was induced, and the shivering threshold was established again after lithotripsy. Results were analyzed using paired or unpaired t-tests. Reduction in the shivering threshold by epidural anesthesia was significantly greater when fentanyl was added to lidocaine than when lidocaine was used alone (mean +/- SD: -0.6+/-0.4 degrees C versus -0.1+/-0.4 degrees C; P < 0.02). We conclude that patients are at increased risk of hypothermia when fentanyl is added to epidural lidocaine. IMPLICATIONS: Fentanyl is often added to lidocaine to improve the quality of epidural blockade and to reduce side effects. However, this study shows that patients are at increased risk of hypothermia when fentanyl is added to lidocaine. PMID- 9728835 TI - Intrathecal, but not intravenous, clonidine reduces experimental thermal or capsaicin-induced pain and hyperalgesia in normal volunteers. AB - Clonidine is approved for intraspinal administration in the treatment of neuropathic cancer pain. Some studies have suggested an analgesic effect after systemic clonidine administration. The purpose of this study was to compare the analgesic effects of intrathecal and IV clonidine with acute noxious stimulation and with hyperalgesia from intradermal capsaicin injection in volunteers. Sixteen healthy volunteers received intradermal injections of capsaicin (100 microg) before and after the IV or intrathecal injection of clonidine 50 or 150 microg in a randomized, double-blind manner. Pain and areas of mechanical hyperalgesia and allodynia were determined at specified intervals. In addition, pain to noxious heat stimulation was determined. The capsaicin injection produced pain, followed by hyperalgesia and allodynia. The intrathecal, but not IV, injection of 150 microg of clonidine reduced capsaicin-induced pain and area of hyperalgesia. Intrathecal clonidine (150 microg) reduced pain to heat stimulation, whereas IV clonidine did not. The groups did not differ in hemodynamic or sedative effects from clonidine. These data support the value of intraspinal administration of clonidine for the treatment of acute pain and of pain states associated with hyperalgesia. Similarly, they suggest that analgesia from the systemic administration of this alpha2-adrenergic agonist, if any, is weak in doses that produce sedation and reduce blood pressure. IMPLICATIONS: To the extent that the experimental pain conditions used in this study reflect those in patients with acute and chronic pain, these data support the spinal rather than IV injection of clonidine for analgesia. PMID- 9728836 TI - A double-blind comparison of ropivacaine, bupivacaine, and mepivacaine during sciatic and femoral nerve blockade. AB - No study has evaluated the efficacy of ropivacaine in peripheral nerve block of the lower extremity. The purpose of this prospective, randomized, double-blind study was to compare ropivacaine, bupivacaine, and mepivacaine during combined sciatic-femoral nerve block. Forty-five ASA physical status I or II patients scheduled for elective hallux valgus repair with thigh tourniquet were randomized to receive combined sciatic-femoral block with 0.75% ropivacaine (ROPI, n = 15), 0.5% bupivacaine (BUPI, n = 15), and 2% mepivacaine (MEPI, n = 15). Time required for onset of sensory and motor block on the operated limb (readiness for surgery) and resolution of motor block, as well as onset of postsurgical pain and time of first analgesic requirement, were recorded. The three groups were similar with regard to demographic variables, duration of surgery, and measured visual analog pain scores. Onset of sensory and motor blockade was similar in Groups ROPI and MEPI and significantly shorter than in Group BUPI (P = 0.002 and P = 0.001, respectively). Resolution of motor block occurred later in Groups ROPI and BUPI than in Group MEPI (P = 0.005 and P = 0.0001, respectively). Duration of postoperative analgesia was significantly longer in Groups ROPI (670+/-227 min) and BUPI (880+/-312 min) compared with Group MEPI (251+/-47 min) (P = 0.0001), with a significant decrease in postoperative pain medication requirements (P < 0.05). We conclude that for sciatic-femoral nerve block, 0.75% ropivacaine has an onset similar to that of 2% mepivacaine and a duration of postoperative analgesia between that of 0.5% bupivacaine and 2% mepivacaine. IMPLICATIONS: Quick onset of block with prolonged postoperative analgesia is an important goal in peripheral nerve blockade. We evaluated the clinical properties of 0.5% bupivacaine, 2% mepivacaine, and 0.75% ropivacaine for sciatic-femoral nerve block and demonstrated that ropivacaine has an onset similar to that of mepivacaine but allows for postoperative analgesia between that of bupivacaine and mepivacaine. PMID- 9728837 TI - Lidocaine concentrations in plasma and cerebrospinal fluid after systemic bolus administration in humans. AB - Preclinical studies suggest that systemic lidocaine acts at the level of the spinal dorsal horn to inhibit hyperalgesia resulting from nerve injury, yet no clinical data are available to support this view. Therefore, we sought to characterize the time course of lidocaine in the plasma and cerebrospinal fluid (CSF) after an IV bolus injection of lidocaine 2 mg/kg in patients scheduled for surgery involving spinal anesthesia. Sixty-five patients were randomly allocated to one of five study groups (n = 13 per group) receiving IV lidocaine before CSF/ plasma sampling at 5, 10, 15, 30, or 60 min. Gas chromatographic analysis of these samples revealed a fast but transient peak (5-15 min) in lidocaine plasma levels (1.7+/-0.16 microg/mL), which declined rapidly thereafter. Only small concentrations of IV lidocaine were found in the CSF (6%- 8% of plasma concentration), but this fraction remained stable from 15 min until termination of the experiment. No statistical correlation was observed between plasma and CSF lidocaine levels. These data suggest that because of the prolonged availability of lidocaine at the spinal dorsal horn level, systemic administration of lidocaine suppresses central sensitization within the spinal cord after nerve injury in humans. IMPLICATIONS: Cerebrospinal fluid concentrations of lidocaine after its systemic bolus delivery in humans indicate that the spinal cord may be the major site of antinociceptive action by this route of drug administration. PMID- 9728838 TI - Intrathecal administration of excitatory amino acid receptor antagonists or nitric oxide synthase inhibitor reduced autotomy behavior in rats. AB - Peripheral nerve injury may produce neuropathic pain. At the spinal cord level, excitatory amino acid receptors play a role in nociceptive signal modulation. N methyl-D-aspartate (NMDA) receptor activation initiates the NO-cGMP pathway and further modulates nociceptive signal. Using the autotomy model, we examined the effect of an NMDA and a non-NMDA receptor antagonist, and nitric oxide synthase inhibitor in the treatment of autotomy behavior in rats. A right-side brachial plexus (C5-T1) transection was performed in all rats. In the treatment groups, MK 801, 1-(4-chlorobenzoyl)-piperazine-2,3-dicarboxylic acid (a non-NMDA antagonist), and L-NG-nitro arginine methyl ester (L-NAME) were infused intrathecally via an osmotic pump for 7 days at doses of 10, 4, and 400 microg/h, respectively. Saline was infused to control animals. Autotomy behavior was observed daily for 8 wk. The incidence of autotomy was 85% in the control group. MK-801, the non-NMDA antagonist, and L-NAME reduced the autotomy incidence to 10%, 20%, and 10% (P < 0.0001), respectively. All treatments delayed the onset of the autotomy behavior from 15+/-4.5 days in the control group to 52+/-3.8, 43+/ 5.6, and 54+/-2.9 (P < 0.001) days in the treatment groups, respectively. The average autotomy scores were also attenuated significantly by these treatments. We conclude that the excitatory amino acid receptors and their intracellular signal messenger NO play a role in deafferentation behavior development. Inhibition of this signaling pathway may be of use for neuropathic pain relief. IMPLICATIONS: The excitatory amino acid receptors and their intracellular signal messenger NO play a role in deafferentation behavior development. Inhibition of this signaling pathway may be of use for neuropathic pain relief. PMID- 9728839 TI - Intrathecal clonidine and fentanyl with hyperbaric bupivacaine improves analgesia during cesarean section. AB - Seventy-eight pregnant women at term, scheduled for elective cesarean section, were enrolled in this multicenter trial to compare the analgesic efficacy and side effect profile of a spinal block with hyperbaric bupivacaine alone (Group B) or combined with 75 microg of clonidine (Group BC) or with clonidine 75 microg and fentanyl 12.5 microg (Group BCF). Intraoperatively, clonidine increased the spread of the sensory block and decreased pain (pain scores 23+/-7 mm vs 17+/-6 and 2+/-1 mm for Group B versus Groups BC and BCF; P < 0.05) and analgesic supplementation. This improved analgesia was best with the clonidine-fentanyl combination (Group BC versus Group BCF; P < 0.05). Postoperative analgesia was prolonged only in Group BCF (215+/-79 min vs 137+/-35 and 183+/-80 min for Group BCF versus Groups B and BC; P < 0.05). Blood pressure and heart rate changes were not significantly different among groups, whereas sedation and pruritus were significantly more frequent in Group BCF. Nausea and vomiting were decreased in Groups BC and BCF. Apgar scores and umbilical artery blood pH were not different among groups. We conclude that adding a small dose of intrathecal clonidine to bupivacaine increases the quality of intraoperative analgesia and decreases pain during cesarean section. Combining clonidine with fentanyl further improved analgesia. IMPLICATIONS: In this study, we demonstrate improved intraoperative spinal analgesia by adding 75 microg of clonidine to bupivacaine; side effects were not increased. The combination of clonidine and fentanyl further improved analgesia but moderately increased sedation and pruritus. PMID- 9728840 TI - The effects of maternal position during induction of combined spinal-epidural anesthesia for cesarean delivery. AB - Combined spinal-epidural anesthesia (CSE) is a popular technique for cesarean delivery. Regional blocks in obstetrics are often performed with the parturient in the sitting position because the midline may be recognized more easily than in the lateral decubitus position. When conventional spinal anesthesia is performed in the sitting position, the patient is placed supine immediately after drug injection. In contrast, when CSE is performed with the woman sitting, there is a delay in assuming the supine position because of epidural catheter placement, which may affect the incidence of hypotension. Healthy women, at term of pregnancy, about to undergo an elective cesarean section under CSE, were randomly assigned to the sitting or lateral recumbent position for initiation of the block. All parturients were given 1000 mL of lactated Ringer's solution in the 15 min preceding induction and an additional 300-500 mL while the actual block was being performed. On completion of the CSE, they were turned to the supine position with left uterine displacement. A second anesthesiologist, blinded to the woman's position during CSE, evaluated the sensory level of anesthesia, maternal heart rate, blood pressure, oxygen saturation, need for ephedrine, and occurrence of nausea and vomiting. Results are expressed as mean +/- SD. Twelve women were studied in the sitting group and 10 were studied in the lateral recumbent group. The severity and duration of hypotension were greater in those parturients who had CSE induced in the sitting (47%+/-7% and 6+/-3 min, respectively) compared with the lateral recumbent position (32%+/-14% and 3+/-2 min, respectively). Women in the sitting group also required twice as much ephedrine (38+/-18 mg) to correct hypotension compared with the other group (17+/ 12 mg). In conclusion, the severity and duration of hypotension were greater when CSE was induced in the sitting compared with the lateral decubitus position. IMPLICATIONS: We studied the induction of combined spinal-epidural anesthesia (CSE) in the sitting versus lateral recumbent positions in healthy women undergoing a scheduled cesarean delivery. The severity and duration of hypotension were greater when CSE was induced in the sitting position. Thus, the position used for induction of CSE should be among the factors considered when there is greater maternal or fetal risk from hypotension. PMID- 9728842 TI - Intrathecal ropivacaine for labor analgesia: a comparison with bupivacaine. AB - Ropivacaine has less potential for central nervous system and cardiovascular toxicity than bupivacaine; in pregnant patients and volunteers, it produces less motor block in equianalgesic doses than bupivacaine. We compared two doses of intrathecal ropivacaine combined with sufentanil with a standard dose of intrathecal bupivacaine plus sufentanil for labor analgesia using a combined spinal-epidural (CSE) technique. In a prospective, randomized, double-blind fashion, 48 patients requesting labor analgesia received either 2.5 mg of intrathecal bupivacaine plus sufentanil 10 microg (B), 2 mg of intrathecal ropivacaine plus sufentanil 10 microg (R2), or 4 mg of intrathecal ropivacaine plus sufentanil 10 microg (R4). Duration of analgesia and side effects, such as motor block, pruritus, hypotension, ephedrine requirements and fetal bradycardia, were recorded. Duration of analgesia (mean +/- SD) was 79+/-30 min for R2, 98+/ 19 min for R4, and 92+/-38 min for B (P = not significant). No differences in motor block or side effects were detected among the groups. We conclude that ropivacaine, when combined with sufentanil, is effective for providing CSE labor analgesia and offers no advantage over bupivacaine in the studied doses. IMPLICATIONS: In this study, we compared a standard dose of intrathecal bupivacaine with sufentanil for combined spinal epidural analgesia with two doses of the new local anesthetic ropivacaine. Both local anesthetics provided similar labor analgesia duration with equivalent side effect profiles in the doses studied. PMID- 9728843 TI - Postanesthesia care unit length of stay: quantifying and assessing dependent factors. AB - Postanesthesia care unit (PACU) monitoring reduces morbidity and is the standard of care for postsurgical patients. PACUs require large nurse to patient ratios, which contributes to the cost of care. Despite the importance and cost of PACU length of stay (LOS), no standards have been established. We performed an observational study of 340 PACU patients to measure actual and medically appropriate PACU LOS (the time required to achieve a medically stable condition for safe PACU discharge), to identify factors related to LOS, and to create a LOS prediction index. Mean (+/- SD) actual LOS was 95+/-43 min, and appropriate PACU LOS was 71+/-37 min. Appropriate PACU LOS predictors were anesthetic time, anesthetic technique, and amount of intraoperative fluids. Actual LOS was >30 min longer than the medically appropriate LOS for 20% (68 of 340) of the patients. Frequent causes of excessive LOS were waiting for physician release or laboratory or radiographic results. Appropriate LOS may be related primarily to anesthetic factors, and nonmedical issues account for a significant amount of PACU LOS. IMPLICATIONS: Most patients are stabilized immediately after surgery in a postanesthesia care unit (PACU) until their discharge to a hospital ward. However, there are no standards for appropriate PACU length of stay (LOS). In this study, we measured actual and appropriate PACU LOSs and evaluated clinical factors that may influence PACU LOS. PMID- 9728841 TI - Effective analgesia after bilateral tubal ligation. AB - Postpartum bilateral tubal ligation is a brief surgical procedure with minimal tissue injury, yet postoperative recovery times and analgesia requirements are often disproportionately large. To evaluate the analgesic efficacy of local anesthetic infiltration, 20 parturients scheduled for elective minilaparotomy and bilateral tubal ligation with either spinal or epidural anesthesia participated in this prospective, randomized, controlled, double-blind trial. All patients received IV metoclopramide 10 mg and ketorolac 60 mg intraoperatively, as well as preincisional infiltration of the infraumbilical skin incision with 0.5% bupivacaine. Infiltration of bilateral uterine tubes and mesosalpinx was performed with either 0.5% bupivacaine (n = 10) or isotonic sodium chloride solution (saline) (n = 10). IV meperidine (25 mg every 3 min as needed) was given to treat pain in the postanesthesia care unit (PACU). The total amount of meperidine administered in the PACU was significantly larger in the saline group than in the bupivacaine group. Pain scores at 30, 45, 60, 75, and 90 min postoperatively and on the seventh postoperative day were significantly lower in the bupivacaine group than in the saline group. During tubal ligation, infiltration of uterine tubes and mesosalpinx with 0.5% bupivacaine significantly enhanced analgesia both in the immediate postoperative setting and on the seventh postoperative day compared with infiltration with sodium chloride. IMPLICATIONS: During bilateral tubal ligation with either spinal or epidural anesthesia, preemptive analgesia using IV ketorolac, IV metoclopramide, and infiltration of the incised skin and uterine tubes with 0.5% bupivacaine allowed 9 of 10 patients to recover with no pain, nausea, vomiting, or cramping and to maintain good analgesia for 7 days postoperatively. PMID- 9728844 TI - Sharps disposal in the operating room: current clinical practices and costs. AB - In the evolving medical environment, fiscal constraints on medical practice are becoming the norm. The new days of austerity have revived interest in the economics of medical practice. Economic measures, however, should not impinge on the quality of patient care. Waste disposal, in particular, is an area without any direct patient benefit but which carries both short- and long-term ecological costs. Much of how we dispose of waste is dictated in the United States by the Joint Commission for the Accreditation of Hospital Organization, Occupational Safety and Health Administration, state regulations, and individual hospital protocols. In an attempt to elucidate the waste in waste management, we examined the use of standard operating room sharp boxes. Full sharp boxes from three different operating sites were randomly saved. Boxes were weighed and opened, and contents were separated into appropriate sharps: loose needles, scalpels, syringes with uncapped needles, and other. Weight and volume assessments were performed on the nonsharps. True sharp values were derived from nonsharps data. Less than 50% of the contents were appropriate for sharps disposal, with empty glass vials constituting the greatest percentage by weight of nonsharps material. We believe that encouraging the appropriate use of sharps boxes is a potential source for savings. IMPLICATIONS: Sharp boxes were randomly saved from university operating rooms and analyzed for content. The full boxes contained 14% appropriate sharps by weight and <50% appropriate sharps by volume. The largest fraction of nonsharps weight was found to be glass. PMID- 9728845 TI - Amino acid-induced thermogenesis to prevent hypothermia during anesthesia is not associated with increased stress response. AB - Intraoperative infusion of amino acids stimulates oxygen uptake with a resulting increase in heat production, especially during emergence from anesthesia. To determine whether the increased thermogenesis in response to amino acid infusion during and after anesthesia induces any additional stress in surgical patients, plasma catecholamines were measured. Fourteen patients aged 44+/-15 yr scheduled for gall bladder surgery during isoflurane anesthesia were studied. Seven patients received an IV amino acid infusion at the rate of 126 mL/h throughout anesthesia, and seven control subjects received equal volumes of nutrient-free saline. At five defined times during and after anesthesia, arterial adrenaline and noradrenaline concentrations were determined. Arterial blood pressure and heart rate were recorded simultaneously. There were considerable variations from baseline in levels of plasma adrenaline and noradrenaline, heart rate, and blood pressure during the study period, but there were no significant differences between the two groups at any time throughout anesthesia and surgery. In conclusion, the augmented thermogenic effect of amino acids during and at emergence from anesthesia is suggested to occur without imposing any additional stress in the surgical patient. IMPLICATIONS: Amino acid infusion during anesthesia has been demonstrated to markedly increase oxygen consumption and heat production. To determine whether this evokes any stress in the patients, we measured plasma catecholamines during anesthesia. We suggest that temperature preservation during anesthesia by an amino acid infusion occurs without additional sympathoadrenal activity. PMID- 9728846 TI - Postoperative nitrogen excretion after amino acid-induced thermogenesis under anesthesia. AB - Amino acid infusions during general anesthesia induce thermogenesis and prevent postoperative hypothermia. The effects of increased heat production during anesthesia on postoperative nitrogen balance have not been examined. Therefore, we studied the effect of perioperative amino acid infusions on postoperative nitrogen excretion in 24 patients scheduled for hysterectomy. Seven volunteers not subjected to anesthesia or surgery were used as awake controls. During isoflurane anesthesia, 8 patients received acetated Ringer's solution, and 16 patients received an IV amino acid mixture, 240 kJ/h, before and during anesthesia. Rectal temperature and energy expenditure were measured. The urinary nitrogen content was calculated from urea, creatinine, and urate the day before surgery and for 4 days postoperatively. Diets were recorded. In anesthetized control patients, postoperative nitrogen excretion was less than preoperative levels. Those patients also experienced the largest decrease in core body temperature during anesthesia (1.7+/-0.1 degrees C). All had postoperative shivering. In the amino acid-treated patients, the temperature decrease during anesthesia was less pronounced (1.0+/-0.1 degrees C; P < 0.001) and postoperative shivering disappeared. In addition, the nitrogen excretion was unchanged postoperatively, perhaps indicating an increase in protein turnover known to generate heat. In conclusion, the increase in heat production induced by amino acids reduced hypothermia, abolished shivering, and attenuated/normalized the postoperative nitrogen saving that occurred in patients who did not receive amino acids. IMPLICATIONS: We compared nitrogen excretion before and after surgery in patients who received a saline or amino acid infusion during isoflurane anesthesia. The increase in heat production induced by amino acids reduced hypothermia, abolished shivering, and attenuated/normalized the postoperative nitrogen saving that occurred in patients who did not receive amino acids. PMID- 9728847 TI - Substantial changes in arterial blood gases during thoracoscopic surgery can be missed by conventional intermittent laboratory blood gas analyses. AB - Substantial and clinically relevant changes in arterial blood gases are likely to occur during thoracoscopic surgery with one-lung ventilation (OLV). We hypothesized that they may be missed when using the conventional intermittent blood gas sampling practice. Therefore, during 30 thoracoscopic procedures with OLV, the sampling intervals between consecutive intermittent laboratory blood gas analyses (BGA) were evaluated with respect to changes of PaO2, PaCO2, and pHa ([H+]) using a continuous intraarterial blood gas monitoring system. Frequency and timing of BGA were based on the clinical judgment of 16 experienced anesthesiologists who were blinded to the continuously measured values. Extreme fluctuations of PaO2 (37-625 mm Hg), PaCO2 (27-56 mm Hg), and pHa (7.24-7.51) were observed by continuous blood gas monitoring. During 63% of all sampling intervals, PaO2 decreased >20% compared with the preceding BGA value, which remained undetected by intermittent analysis. In 10 patients with a continuously measured minimal PaO2 value < or = 60 mm Hg, the preceding BGA overestimated this minimal PaO2 by > 47%. Correspondingly, PaCO2 increases of > 10% were observed in 35% of all sampling intervals, and [H+] increases of > 10% were observed in 24% of all sampling intervals. Because these blood gas changes were not reliably detected by using noninvasive monitoring and their magnitude is not predictable during OLV, intermittent BGA with short sampling intervals is warranted. In critical cases, continuous blood gas monitoring may be helpful. IMPLICATIONS: The magnitude of blood gas changes during thoracoscopic surgery with one-lung ventilation is not predictable and not reliably detected by noninvasive monitoring. Using a continuous intraarterial blood gas monitoring device, we demonstrated that intermittent laboratory blood gas analysis with short sampling intervals is warranted to detect arterial hypoxemia. PMID- 9728849 TI - The laryngeal mask airway reliably provides rescue ventilation in cases of unanticipated difficult tracheal intubation along with difficult mask ventilation. AB - In 1995, our department of anesthesiology established an airway team to assist in treating unanticipated difficult endotracheal intubations and an airway quality improvement (QI) form to document the use of emergency airway techniques in airway crises (laryngeal mask airway [LMA], flexible fiberoptic bronchoscopy, retrograde intubation [RI], transtracheal jet ventilation [TTJV], and cricothyrotomy). Over a 2-yr period, team members and staff anesthesiologists completed airway QI forms to document the smallest peripheral SpO2 during an airway crisis, the number of direct laryngoscopies (DL) performed before using an emergency airway technique, and the emergency airway technique that succeeded in rescue ventilation. Team members agreed to use the LMA as the first emergency airway technique to treat the difficult ventilation/difficult intubation scenario. A SpO2 value < or =90% during mask ventilation defined difficult ventilation. Inability to perform tracheal intubation by DL defined difficult intubation. An increase in the SpO2 value >90% defined rescue ventilation. Review of airway QI forms from October 1, 1995 until October 1, 1997 revealed 25 cases of difficult ventilation/difficult intubation. Before airway rescue, the median SpO2 was 80% (range 50%-90%), and there were four median attempts at DL (range one to nine). The LMA had a success rate of 94% (95% confidence interval [CI] 77 100). Flexible fiberoptic bronchoscopy, TTJV, RI, and surgical cricothyrotomy had success rates of 50% (95% CI 0-100), 33% (95% CI 0-100), 100% (95% CI 37-100), and 100% (95% CI 37-100), respectively. LMA insertion as the first alternative airway technique was useful in dealing with unanticipated instances of simultaneous difficulty with mask ventilation and tracheal intubation. IMPLICATIONS: Twenty-five cases of simultaneous difficulty with mask ventilation and tracheal intubation occurred after the induction of general anesthesia during the study period. The laryngeal mask was used in 17 cases, and it provided rescue ventilation without complication in 94% of these cases (95% confidence interval 77-100). PMID- 9728848 TI - The effects of body mass on lung volumes, respiratory mechanics, and gas exchange during general anesthesia. AB - We investigated the effects of body mass index (BMI) on functional residual capacity (FRC), respiratory mechanics (compliance and resistance), gas exchange, and the inspiratory mechanical work done per liter of ventilation during general anesthesia. We used the esophageal balloon technique, together with rapid airway occlusion during constant inspiratory flow, to partition the mechanics of the respiratory system into its pulmonary and chest wall components. FRC was measured by using the helium dilution technique. We studied 24 consecutive and unselected patients during general anesthesia, before surgical intervention, in the supine position (8 normal subjects with a BMI < or = 25 kg/m2, 8 moderately obese patients with a BMI >25 kg/m2 and <40 kg/m2, and 8 morbidly obese patients with a BMI > or = 40 kg/m2). We found that, with increasing BMI: 1. FRC decreased exponentially (r = 0.86; P < 0.01) 2. the compliance of the total respiratory system and of the lung decreased exponentially (r = 0.86; P < 0.01 and r = 0.81; P < 0.01, respectively), whereas the compliance of the chest wall was only minimally affected (r = 0.45; P < 0.05) 3. the resistance of the total respiratory system and of the lung increased (r = 0.81; P < 0.01 and r = 0.84; P < 0.01, respectively), whereas the chest wall resistance was unaffected (r = 0.06; P = not significant) 4. the oxygenation index (PaO2/PAO2) decreased exponentially (r = 0.81; P < 0.01) and was correlated with FRC (r = 0.62; P < 0.01), whereas PaCO2 was unaffected (r = 0.06; P = not significant) 5. the work of breathing of the total respiratory system increased, mainly due to the lung component (r = 0.88; P < 0.01 and r = 0.81; P < 0.01, respectively). In conclusion, BMI is an important determinant of lung volumes, respiratory mechanics, and oxygenation during general anesthesia with patients in the supine position. IMPLICATIONS: The aim of this study was to investigate the influence of body mass on lung volumes, respiratory mechanics, and gas exchange during general anesthesia. PMID- 9728850 TI - The effect of fresh gas flow and anesthetic technique on the ability to control acute hemodynamic responses during surgery. AB - We evaluated the effect of the fresh gas flow (FGF) rate and the anesthetic technique on the ability to control the acute hyperdynamic response to a specific surgical stimulus during surgery in 90 consenting ASA physical status I-III patients undergoing lower abdominal procedures. After the administration of midazolam 2 mg IV, anesthesia was induced in all patients with propofol 1.5 mg/kg IV and fentanyl 1 microg/kg IV and was initially maintained with desflurane or isoflurane, 0.7 minimum alveolar anesthetic concentration, at total FGF rates of either 1 or 3 L/min. In response to the surgical stimulation of skin incision and retropubic dissection, an increase in mean arterial pressure (MAP) >20% above the preincision baseline MAP value provoked a stepwise increase in the inspired concentration of the volatile anesthetic or the IV administration of a variable rate infusion of esmolol. At both FGF rates, the acute hemodynamic response to surgical stimulation was more efficiently treated by increasing the inspired concentration of desflurane than isoflurane. At 1 L/min, the average time to control the increase in MAP was significantly shorter with desflurane (17+/-12 min) compared with isoflurane (29+/-16 min), with 60% of the patients in the isoflurane group requiring rescue therapy. When an esmolol infusion was used to control the increase in MAP, supplementation with fentanyl was required in 40% and 53% of patients anesthetized with desflurane and isoflurane, respectively. In conclusion, desflurane provided more rapid and reliable control of acute hemodynamic responses to surgical stimulation than isoflurane or esmolol when the volatile anesthetics were administered at low FGF rates. IMPLICATIONS: At low fresh gas flow rates (1 L/min), desflurane more successfully and rapidly controlled the acute hemodynamic responses to painful surgical stimuli than isoflurane. PMID- 9728851 TI - Esmolol potentiates reduction of minimum alveolar isoflurane concentration by alfentanil. AB - Esmolol, a short-acting beta1-receptor antagonist, decreases anesthetic requirements during propofol/N2O/morphine anesthesia. This study was designed to determine whether esmolol affects the volatile anesthetic (isoflurane) required to prevent movement to skin incision in 50% patients (minimum alveolar anesthetic concentration [MAC]) with or without an additional opioid (alfentanil). One hundred consenting adult patients were randomly divided into five treatment groups: isoflurane alone (I), I with continuous large-dose (250 microg x kg(-1) x min(-1)) esmolol (E), I with alfentanil (effect site target of 50 ng/mL) via a continuous computer-controlled infusion (A), A plus continuous small-dose (50 microg x kg(-1) x min(-1)) esmolol (A1), or A plus large-dose esmolol (A2). Anesthesia was induced via a face mask, and steady-state target end-tidal isoflurane concentrations were maintained before incision. The MAC of isoflurane alone was 1.28% +/- 0.13%. Large-dose esmolol did not significantly alter the isoflurane MAC (1.23% +/- 0.14%). Alfentanil alone significantly decreased isoflurane MAC by 25% (0.96% +/-0.09%). Adding small-dose esmolol did not further decrease MAC with alfentanil (0.96% +/- 0.13%). However, large-dose esmolol significantly decreased isoflurane MAC with alfentanil (0.74% +/- 0.09%). Esmolol and alfentanil both significantly reduced the increases in heart rate and mean arterial pressure associated with endotracheal intubation and incision. The mechanism of this effect is unknown. IMPLICATIONS: Most anesthetic techniques rely on a balance of several highly selective medications. The current results define a new anesthetic-sparing effect when volatile anesthetic, analgesic, and beta-adrenergic blocking drugs are combined. PMID- 9728852 TI - Rapid onset of ulnar nerve dysfunction during transient occlusion of the brachial artery. AB - Perioperative ulnar neuropathy is a complication that occurs even in patients who seem to be appropriately padded and positioned. The disproportionately high incidence of postoperative ulnar nerve injury compared with the median and radial nerves has largely been attributed to its vulnerability to compression or stretch at the cubital tunnel. Some clinical and laboratory evidence suggests that compromise of perfusion to the upper extremity may also play a role in this complication. To determine whether the ulnar nerve is more sensitive to ischemia of the upper extremity, we studied 10 men during general anesthesia. Somatosensory evoked potentials of the radial, median, and ulnar nerves were simultaneously recorded during general anesthesia with the brachial artery occluded proximal to the cubital fossa. All three nerves showed rapid changes in signal amplitude in response to occlusion of the brachial artery, but the amplitude of the ulnar nerve was affected earlier and to a greater degree. Compared with the median nerve, the change in ulnar nerve signal amplitude during ischemia was significantly greater after 4 min (P = 0.002). This trend persisted at 6 and 8 min (P = 0.008). At 4, 6, and 8 min of ischemia, the ulnar nerve likewise showed a greater decrease in amplitude compared with the radial nerve, with corresponding P values of 0.015, 0.008, and 0.008. We conclude that the ulnar nerve is more sensitive to ischemia of the upper extremity compared with the radial and median nerves. In addition to its increased vulnerability at the elbow, compromise of arterial flow may contribute to some cases of postoperative ulnar neuropathy. IMPLICATIONS: Postoperative ulnar neuropathy is thought to result from compression or stretch of the ulnar nerve at the elbow. However, patients may sustain this complication despite careful padding and positioning. This study suggests that the ulnar nerve may also be unusually sensitive to decreases in blood supply to the arm. Care should not only to properly position and pad the elbows, but also to ensure adequate perfusion of the upper extremities. PMID- 9728853 TI - Oral antihistamines reduce the side effects from rapid vancomycin infusion. AB - Rapid infusion of vancomycin causes histamine-mediated side effects, hypotension, and rash, known as "red man syndrome." In this prospective, randomized, double blind, placebo-controlled study, we examined the ability of oral antihistamines to attenuate three clinical end points: rash, hypotension, and vancomycin discontinuation, and we compared these findings with those of a similar study using IV antihistamines. Patients (ASA physical status I-III) who required vancomycin prophylaxis for elective arthroplasty received either oral antihistamines (diphenhydramine < or = 1 mg/kg and cimetidine < or = 4 mg/kg, n = 20) or placebo (n = 10) 1 h before rapid vancomycin infusion (1 g over 10 min). The vancomycin infusion was discontinued if the mean arterial blood pressure decreased by > or = 20% or if itching was intolerable for the patient. Clinically significant hypotension developed in no treated patients, compared with five (50%) patients in the placebo group (P = 0.001). Rapid infusion was stopped for one treated patient (5%) and for five (50%) patients in the placebo group (P = 0.004). Incidence (P = 0.011) and severity of rash (P = 0.015) were also reduced in treated patients. Peak histamine levels were increased but were similar for patients in both groups (mean +/- SD, 1.9+/-2.5 vs 1.6+/-2.4 ng/mL; P = 0.75). Oral antihistamines were as effective as IV antihistamines. In conclusion, oral H1 and H2 antihistamine pretreatment is a practical, safe, and inexpensive option to attenuate histamine-mediated side effects associated with rapid vancomycin infusion. IMPLICATIONS: Clinicians often must administer vancomycin faster than the 1-h recommended time, which can cause "red man syndrome" (rash, itching, hypotension). Our randomized, double-blind, placebo-controlled study showed that oral H1 and H2 antihistamine pretreatment significantly reduced the histamine related side effects of rapid vancomycin infusion. PMID- 9728854 TI - The effects of intraoperative intravenous clonidine on gastric intramucosal PCO2. AB - To investigate the effects of clonidine given as an anesthetic adjunct on splanchnic perfusion, we determined intramucosal gastric PCO2 using gastric tonometry in 60 patients scheduled for large intestine surgery. After induction of anesthesia, patients were randomly assigned to four groups. Patients in Group 1 received an IV infusion of sufentanil (0.2 microg x kg(-1) x h(-1)); patients in Group 2 received an IV infusion of clonidine (4 microg/kg in 20 min followed by 2 microg x kg(-1) x h(-1)); patients in Group 3 received an IV infusion of ketamine (0.5 mg/kg followed by 0.25 mg x kg(-1) x h(-1)); patients in Group 4 received an epidural infusion of bupivacaine (7 mL of 0.5% followed by 5 mL/h of 0.25%). Gastric intramucosal PCO2 was assessed immediately before skin incision and every hour during surgery using a nasogastric tube. A last measurement was taken after skin closure. An arterial sample was collected simultaneously to measure arterial PCO2. Oxygen consumption (VO2/min) was continuously recorded. Gastric intramucosal PCO2 significantly increased during surgery in all groups independent of the anesthetic technique considered (P < 0.01) and was not related to metabolic changes or blood pressure variations. This increase, however, remains in the physiologic range. In conclusion, our results demonstrate that clonidine given as an anesthetic adjutant at the dose used has no deleterious effect on splanchnic perfusion during colonic surgery. IMPLICATIONS: IV clonidine given as an anesthetic adjunct at a dose of 4 microg/kg in 20 min, followed by 2 microg x kg(-1) x h(-1), has no deleterious effect on splanchnic perfusion during colonic surgery. PMID- 9728855 TI - The effects of verapamil and diltiazem pretreatment on potassium release in dogs after the administration of succinylcholine. AB - Verapamil exacerbates the increase in serum potassium after a large-dose potassium infusion or after the IV administration of succinylcholine. We conducted a study in 12 canines conditioned for 30 days acting as their own controls. The canines had chronic tracheotomies and carotid loops performed 2 wk before the experiment. Control canines were given 1 mg/kg succinylcholine at Time 0. Blood samples were analyzed for potassium at 0, 1, 3, 5, 10, 15, 30, and 60 min. One week later, the dogs received 0.15 mg/kg of either verapamil or diltiazem, followed by a 5.6-microg x kg(-1) x min(-1) 10-min infusion of the same drug. The animals were then given a bolus dose of succinylcholine 1 mg/kg, and the blood potassium was analyzed as before. There was no significant difference in the potassium concentration before the succinylcholine injection (Time 0) between the study groups. The canines pretreated with verapamil had a significantly greater increase (24% +/- 8%) in potassium concentration than the control canines (14% +/- 6%) 15 min after succinylcholine administration. There was no difference between the potassium concentrations of the diltiazem pretreated canines and the control group at any time point. Therefore, diltiazem pretreatment does not significantly influence potassium regulation after a succinylcholine injection, whereas verapamil pretreatment has measurable hyperkalemic effects. IMPLICATIONS: Succinylcholine is a drug that causes blood potassium to increase. Potassium influences heart rhythm. Verapamil and diltiazem are drugs used for angina heart pain. We used dogs to determine the effect of verapamil or diltiazem on the blood's potassium after an injection of succinylcholine and found that verapamil had the greatest effect. PMID- 9728856 TI - Propofol inhibits human neutrophil functions. AB - Neutrophils play important roles in the antibacterial host defense mechanism and in the pathogenesis of tissue injury. Propofol has been reported to impair the production of reactive oxygen species from neutrophils. We examined the effect of propofol (2,6-diisopropylphenol), at clinically relevant concentrations and at 10 and 100 times this concentration, on several aspects of human neutrophil functions using an in vitro system. Propofol significantly inhibited chemotaxis, phagocytosis, and reactive oxygen species (ROS) (O2-, H2O2, OH) production of neutrophils in a dose-dependent manner. At clinically relevant concentrations, propofol suppressed these neutrophil functions, but it did not decrease ROS generation by the cell-free (xanthine-xanthine oxidase) system. Increase in intracellular calcium concentrations in neutrophils stimulated by N-formyl-L methionyl-L-leucyl-L-phenylalanine was dose-dependently attenuated by propofol. This decreasing effect on [Ca2+]i in neutrophils may represent one of the mechanisms responsible for the inhibition of neutrophil functions by propofol. IMPLICATIONS: Neutrophils play a pivotal role in the antibacterial host defense system and tissue injury. We found that at clinically relevant concentrations, propofol impaired neutrophil functions. Further studies may determine whether this impairment, observed in vitro, leads to clinical immunological suppression. PMID- 9728857 TI - Halothane and isoflurane alter calcium dynamics in rat cerebrocortical synaptosomes. AB - An increase in synaptosomal Ca2+ triggers neurotransmitter release and volatile anesthetics have been shown to inhibit neurotransmitter release by inhibition of Ca2+ entry. We have examined the effect of isoflurane and halothane on the kinetics of increase and decrease of Ca2+ in rat cerebrocortical synaptosomes ([Ca2+]in). We have also used specific Ca2+ antagonists to examine the role of L , N-, and P-type Ca2+ channels. Synaptosomal [Ca2+]in was measured spectrofluorometrically using fura-2 as a Ca2+ reporter; Ca2+ transients were initiated by depolarization with 40 mM KCl. We found that < or = 1 minimum alveolar anesthetic concentration halothane and isoflurane decreased peak [Ca2+]in by approximately 40%, that both anesthetics decreased the rate of [Ca2+]in increase and decrease, that specific voltage-dependent calcium channel antagonists had little effect on peak or plateau [Ca2+]in, and that the volatile anesthetics increased the permeability of synaptosomal membranes to Ca2+. These results suggest that the volatile anesthetics, at clinically relevant concentrations, can alter Ca2+ homeostasis in the synapse. IMPLICATIONS: Clinically relevant concentrations of halothane and isoflurane markedly depress K+-evoked increases in rat cerebrocortical synaptosomal calcium (Ca2+) unrelated to L-, N-, and P-type voltage-dependent calcium channels and increase the Ca2+ permeability of the synaptosomal membrane. These changes in Ca2+ dynamics could have profound effects on Ca2+ signaling in the synapse. PMID- 9728858 TI - The use of intraoperative nitrous oxide leads to postoperative increases in plasma homocysteine. AB - Hyperhomocysteinemia is an independent risk factor for coronary artery and cerebrovascular disease, but its significance in the perioperative period is unknown. Nitrous oxide inhibits methionine synthase, which aids in the conversion of homocysteine to methionine. In this prospective, controlled, randomized study, we determined the effect of intraoperative nitrous oxide exposure on postoperative plasma homocysteine concentrations. Twenty ASA physical status I III patients, aged >18 yr, presenting for elective craniotomy, were randomized to receive general anesthesia with or without nitrous oxide (inspired nitrous oxide >50%). Plasma was sampled before the induction of anesthesia, on arrival in the postanesthesia care unit (PACU) after discontinuation of nitrous oxide, and 24 h after induction. There was a significant increase (22.6+/-11.4 vs 13.0+/-4.7 micromol/L; P = 0.0038 for postoperative versus preinduction values) in plasma homocysteine concentrations in the nitrous oxide group on arrival in the PACU and for 24 h. In the nonnitrous oxide group, mean plasma homocysteine concentrations did not change (9.5+/-1.9 vs 9.8+/-1.6 micromol/L; P = 0.86 for postoperative versus preinduction values). The change in plasma homocysteine concentrations in the nitrous oxide group was significantly different from that in the nonnitrous group (P = 0.0031). We conclude that the use of intraoperative nitrous oxide leads to significant increases in perioperative plasma homocysteine concentrations. IMPLICATIONS: Short-term exposure to nitrous oxide led to significant increases in plasma homocysteine. Further investigations are required to determine the clinical significance of this change. PMID- 9728859 TI - Widespread application of topical steroids to decrease sore throat, hoarseness, and cough after tracheal intubation. PMID- 9728860 TI - Safety of a new lightwand device (Trachlight): temperature and histopathological study. PMID- 9728861 TI - A randomized, double-blind comparison of rocuronium, d-tubocurarine, and "mini dose" succinylcholine for preventing succinylcholine-induced muscle fasciculations. PMID- 9728862 TI - Statistical analysis of total labor pain using the visual analog scale and application to studies of analgesic effectiveness during childbirth. PMID- 9728863 TI - Laryngeal mask airway facilitates awake fiberoptic intubation in a patient with severe oropharyngeal bleeding. PMID- 9728864 TI - Traumatic esophageal perforation resulting from endotracheal intubation. PMID- 9728865 TI - Damaged Univent tubes. PMID- 9728866 TI - Capnography-guided nasotracheal intubation of a patient with a difficult airway and unwanted respiratory depression. PMID- 9728867 TI - Hemodynamic responses to electroconvulsive therapy in a hypertensive patient with end-stage pulmonary fibrosis. PMID- 9728868 TI - Allocating subspecialist pediatric anesthesiologists: when to say no. PMID- 9728870 TI - Neither obsession nor distraction: words must be chosen well. PMID- 9728869 TI - Interpretation of nonpulsatile arterial pressure-flow relations during cardiopulmonary bypass. PMID- 9728871 TI - Use of plastic rod/sleeve combination to facilitate double- to single-lumen tracheal tube exchange in patients with difficult glottic visualization. PMID- 9728872 TI - Interluminal connection in a failed central venous line. PMID- 9728873 TI - Epidural analgesia and intravenous patient-controlled analgesia result in similar rates of myocardial ischemia after aortic surgery. PMID- 9728874 TI - Occult air leak of an endotracheal tube. PMID- 9728875 TI - Preferential effect of nitroglycerin on large microvessels. PMID- 9728876 TI - Epidural analgesia and chronic backache. PMID- 9728877 TI - Case of frozen thiopental. PMID- 9728878 TI - Pharyngoscopic views. PMID- 9728879 TI - Introduction to microscopic research of silver halides and related dispersed systems. PMID- 9728880 TI - Transmission electron microscopy studies of (111) twinned silver halide microcrystals. AB - The present paper covers the results of different transmission electron microscopy studies on silver halide microcrystals. Pure AgBr as well as core shell AgBr-AgBrI crystals are investigated. In the former parallel and non parallel twinning modes yielding tabular and needle- or tetrahedral-shaped microcrystals, respectively, are discussed. Also the short-range-order of Ag+ interstitials around the twin planes as determined from diffuse intensity in reciprocal space is described. The latter yields a technique to determine the variant in which dislocations are located in certain core-shell microcrystals. The introduction of iodine also results in the presence of inclined stacking faults in the shell and of long-range iodine ordering on the crystal surface. PMID- 9728881 TI - XPS and STM studies of pseudo-AgX monolayers organized on Ag(111) and Au/Ag(111) surfaces. AB - Simple and well-defined model AgX surfaces are useful to illuminate a variety of surface-related phenomena associated with AgX microcrystals in a different perspective with fewer experimental impediments. From this viewpoint, we discuss a couple of pseudo-AgX monolayers that can be organized on a highly planar sputter-grown Ag(111) or ultrathin-Au-covered Ag(111) film. The monolayer structures have been satisfactorily characterized by combination of XPS and STM methods. In particular, the monolayer AgX formed on Au/Ag(111) proved to have an almost identical structure with the bilayer AgX(111) atomic configuration, thus serving as a superior monolayer model of AgX or pseudo 2D ionic crystal for deeper understanding of the processes involving the AgX surfaces. PMID- 9728882 TI - Combined characterization of composite tabular silver halide microcrystals by cryo-EFTEM/EELS and cryo-STEM/EDX techniques. AB - The combination of cryo-energy filtering transmission electron microscopy (EFTEM)/electron spectroscopic diffraction (ESD)/electron energy-loss spectroscopy (EELS) and cryo-energy-dispersive X-ray (EDX) analysis in the scanning transmission (STEM) and scanning (SEM) modes was applied for the characterization of composite tabular Ag(Br,I) microcrystals. A low-loss fine structure in EEL spectra between 4 and 26 eV was attributed to excitons and plasmons possibly superimposed with interband transitions and many-electron effects. The contrast tuning under the energy-filtering in the low-loss region was used to image the crystal morphology, defect structure (random dislocations and ?111? stacking faults) and bend and edge contours as well as electron excitations in the microcrystals. Sharp extra reflections at commensurate positions in between the main Bragg reflections and diffuse honeycomb contours in ESD patterns of the microcrystals taken near the [111] zone were assigned to the number of defects in the shell region parallel to the grain edges and polyhedral clusters of interstitial silver cations, respectively. The imaginary part of the energy-loss function, Im (-1/epsilon), and the real and imaginary parts, epsilon1 and epsilon2, of the dielectric permittivity were determined by means of a Kramers-Kronig analysis. An assignment of exciton peaks based on calculations of electronic band structure of silver bromide is proposed. Inner-shell excitation bands of silver halide were detected in line with EDX-analyses. The energy-loss near-edge structure (ELNES) of the AgM4,5-edge governed by spin-orbital splitting between the 3d3/2- and 3d5/2-states has been evaluated. Combined silver and halide distributions were obtained by a three-window method (EFTEM) and by EDX/STEM including area mapping and line profiling of iodide. PMID- 9728883 TI - Morphology and structure of photosensitive dye J-aggregates adsorbed on AgBr microcrystals grown in gelatin. AB - Though the cyanine dye J-aggregates carry the role to sense the exposing light in the silver halide photographic system, little research on the morphology of the aggregates in adsorption has been made with modern surface analytical methods. In this paper, we describe the size, epitaxy, multi-layered array formation, nucleation and preferential adsorption, and irregular distribution of population between particles and the segregation on a particle, of J-aggregates adsorbed on AgBr grown in gelatin. We employed cathodoluminescence microscopy, low energy high resolution scanning electron microscopy, and atomic force microscopy. Dye molecules aggregate together near the surface of AgBr and adsorb on the surface. The growth of adsorbed aggregates is controlled by the diffusion of dye molecules from the surrounding solution. The population of J-aggregates adsorbed on an AgBr particle varies from almost none to full coverage. Each aggregate is about (20 30) x (30-50) nm in size and is 2.1 nm thick for thiacarbocyanine with sodium ion, 1.04 nm for thiacarbocyanine with tosyl ion, and 0.5 nm for an oxacarbocyanine. The aggregates connect their longer edges to each other to form arrays, and the arrays build up multi-layered stacks. The arrays align parallel and segregate to form terraces. The longer edges of J-aggregates align along [210] on AgBr (100) or [632] on AgBr (111). PMID- 9728884 TI - Investigation of growth of AgX tabular crystals. AB - The process of tabular crystal formation during physical ripening of fine AgX (X=Br,I) emulsions is investigated. The tabular crystal growth is realized simultaneously via two mechanisms: ionic and coalescent. The relative contribution of those mechanisms to the tabular crystal growth is determined by the conditions of the physical ripening. It is possible to control the size and shape uniformity of the final AgX tabular crystals by adjusting the relative contributions of these mechanisms. PMID- 9728885 TI - Mass crystallization of silver chloride microcrystals. AB - In this work some problems on silver chloride isometric and flat crystals crystallization are considered. It is shown that the synthesis of monosize isometric microcrystals should proceed in the absence of a silver chloride solvent. The flat silver chloride microcrystals, in the absence of growth modifiers, are formed by a coalescent mechanism. PMID- 9728886 TI - Research of new AgX crystal forms by transmission electron microscopy. AB - The research of AgX (X=Cl, Br) crystals grown from ammonia solution as a function of X- supersaturation, gelatin concentration, temperature, and use of growth modificators, has been enhanced by data from transmission electron microscopy. The study resulted in preparing AgBr thin films with the most developed ?100? side of rombododecahedron. The size of the crystals was 5 x 15 mm. The thickness was several microm. The new forms of AgBr and AgCl microcrystals were obtained from a high concentrated solution of the corresponding salts in NH4OH. These microcrystals were named "X," 'Y' crystals [AgBr] and stereostructures [AgCl] deriving from their external shapes. As a result of the variation of crystallization parameters, six various morphological types of stereostructures, distinguishable by the size of a central element in relation to external elements and a length of a binding element, were seen. Experiments on photolysis of the stereostructures at the substrate temperature 25-200 degrees C were carried out in vacuum. Exposure to mercury lamp light resulted in characteristic location of large particles of photolytic silver on the stereostructure surface, depending on substrate temperature. PMID- 9728887 TI - Electron microscope characterization of AgBr heterojunctions with silver carboxylates and their influence on the morphology of developed silver particles in thermally developed photomaterials. AB - Silver halide crystals formed during in situ treatment of silver stearate crystals with various halidizing agents are observed by scanning and transmission electron microscopy to form on the lateral edges of the silver carboxylate crystals. The location of the silver halide phase on the crystal edge is dictated by the anisotropic structure of the silver stearate crystal lattice, specifically, the layered structure in which silver ion layers are separated by long-chain hydrocarbon groups. The formation of AgBr on the lateral faces of these crystals is proposed to be typical not only of the formation of silver halide on silver stearate but also for all silver carboxylates of the general formula [AgCnH2(n-1)O2]2 when the crystals of these silver carboxylates have anisotropic, layered structures. The silver bromide/silver carboxylate heterojunction in an in situ system has been clearly observed by transmission electron microscopy. The heterojunction is comprised of a distorted silver carboxylate lattice, which accommodates the misalignment between the AgBr and [Ag(O2CR)]2 crystal lattices. The nature of heterojunction between the AgBr and the silver carboxylate when the AgBr is prepared separately from the preparation of the silver carboxylate differs from the in situ heterojunction. In this case, a layered compound, proposed to have a Ag1-xNaxSt composition, forms between the AgBr and the silver stearate which is a unique feature of this interface. The differences in the structure of interfaces formed between the silver halide and the silver fatty acid complex result in different silver particle morphologies during thermal development of exposed photothermographic films. The developed silver is generally filamentary when the photothermographic material contains silver halide prepared by the in situ exchange reaction between silver carboxylate and a brominating agent. If the photothermographic material is prepared from previously synthesized silver halide crystals, the preformed AgBr route, the developed silver generally crystallizes as dendritic crystals. PMID- 9728888 TI - Human malaria: segregation analysis of blood infection levels in a suburban area and a rural area in Burkina Faso. AB - The genetic control of blood infection levels in human malaria remains unclear. Case control studies have not demonstrated a strong association between candidate genes and blood parasite densities as opposed to surveys that have focused on severe malaria. As an alternative approach, we used segregation analyses to determine the genetic control of blood parasitemia. We surveyed 509 residents (53 pedigrees) in a rural area and 389 residents (41 pedigrees) in an urban area during 18 months. Each family was visited 20 times and 28 times in the urban area and in the rural area; the mean number of parasitemia measurements per subject was 12.1 in the town and 14.9 in the village. The intensity of transmission of Plasmodium falciparum was 8-fold higher in the rural area than in the urban area. Using the class D regressive model for both populations, we found that blood parasite densities were correlated between sibs. We obtained strong evidence for a major effect, but we found that the transmission of this major effect was not compatible with a simple Mendelian model, suggesting a more complex mode of inheritance. Moreover, there was a strong interaction between major effect and age, suggesting that the influence of the putative major gene may be more prominent in children than in adults. Further nonparametric linkage studies, such as sib pair analysis, that focus on children would help us better understand the genetic control of blood infection levels. PMID- 9728889 TI - Testing for contributions of mitochondrial DNA mutations to complex diseases. AB - Several complex disorders are suspected of being associated with mitochondrial DNA (mtDNA) mutations. We studied the statistical properties of a test based on proband-relative pairs to identify potential mtDNA mutation involvement in a complex disorder. The test compares the recurrence risk of relatives of probands along the mitochondrial lineage with that of relatives along the nonmitochondrial lineage. If mtDNA mutations are involved, the recurrence risk will be higher among relatives in the mitochondrial lineage. The form of the test is independent of the assumed models of inheritance and interaction of the nuclear autosomal mutations with mtDNA mutations. The power of the test, however, differs among the different models and by the type of proband-relative pairs used in the test. We considered heterogeneity models with and without phenocopies, a three-state heteroplasmic mtDNA transmission model, and a multiplicative epistasis model. Under the heterogeneity model, the power of the test increases as the relationship between the proband and the relative becomes more distant. Under the multiplicative epistasis model, the power of the test decreases as the relationship between the proband and the relative becomes more distant. PMID- 9728890 TI - Parental genotype reconstruction: applications of haplotype relative risk to incomplete parental data. AB - Intended to resolve the problem of constructing a matched population-based control sample, haplotype relative risk techniques frequently suffer from loss of power for late-onset diseases due to unavailability of parental genotypes that are required to form parent-offspring pairs. However, much of this missing information can be reconstructed using the genotypes of the affected individual's siblings. For some cases, an estimate of the marker-allele frequencies for controls is needed. We have developed a technique based upon this idea and call it PGR (Parental Genotype Reconstruction). PGR represents a combination of the internal controls used by haplotype relative risk and controls based upon estimated allele frequencies. Although the latter is just what haplotype relative risk was designed to avoid, PGR has the potential to substantially boost power to detect linkage disequilibrium, for a relatively small increase in type I error. This performance is reasonably robust with respect to errors in the estimated allele frequencies, as we demonstrate by numerical simulation. A PGR software package has been written in FORTRAN and is available from Dr. Martin's web site at http://electro.psi.vcu.edu/rmartin/. PMID- 9728891 TI - Two-locus developments of the weighted pairwise correlation method for linkage analysis. AB - Different studies of complex traits assumed to be influenced by two unliked loci found that two-locus linkage analysis is more powerful than the classical one locus strategy. The Weighted Pairwise Correlation (WPC) approach is a nonparametric method for linkage analysis that has the advantage to analyze any kind of phenotypes and to consider extended pairs of relatives. In this report, we propose different two-locus extensions of the WPC method based on an additive or a multiplicative effect of two unlinked marker loci on the phenotype. Both methods and their corresponding statistics are easily derived from the classical WPC approach. Compared to the additive model, the multiplicative approach, which can be understood as a statistical interaction effect of the two markers, does not need to specify any additional parameter and allows one to test both the global effect of the two markers (T(AB)test) and the effect of one marker, e.g., B, taking into account the effect of the other, A (T(AB/A) test). When compared to classical one-locus tests by means of simulations, two-locus tests have comparable 0.05 type I error and are more powerful. In particular, tests based on the multiplicative approach appear to be quite interesting in addition to single locus tests to detect the combined role of two markers (T(AB)), or to investigate the role of a marker taking into account a known linked marker (T(AB/A)), especially when these markers have complex effects on the phenotype (e.g., statistical interaction). PMID- 9728892 TI - Alternative test for linkage between two loci. PMID- 9728893 TI - The contribution of intestinal and hepatic CYP3A to the interaction between midazolam and clarithromycin. AB - OBJECTIVE: To assess the relative contribution of intestinal and hepatic CYP3A inhibition to the interaction between the prototypic CYP3A substrates midazolam and clarithromycin. METHODS: On day 1, 16 volunteers (eight men and eight women; age range, 20 to 40 years; weight range, 45 to 100 kg) received simultaneous doses of midazolam intravenously (0.05 mg/kg over 30 minutes) and orally (4 mg of a stable isotope, 15N3-midazolam). Starting on day 2, 500 mg clarithromycin was administered orally twice daily for 7 days. On day 8, intravenous and oral doses of midazolam were administered 2 hours after the final clarithromycin dose. Blood and urine samples were assayed for midazolam, 15N3-midazolam, and metabolites by gas chromatography-mass spectrometry. RESULTS: There was no significant (p > 0.05) difference in the urinary excretion of 1'-hydroxymidazolam after intravenous and oral dosing on day 1 or day 8, indicating that the oral dose was completely absorbed into the gut wall. The oral clearance of midazolam was found to be significantly greater in female subjects (1.9 +/- 1.0 versus 1.0 +/- 0.3 L/hr/kg; p < 0.05) than in male subjects but not systemic clearance (0.35 +/- 0.1 versus 0.44 +/- 0.1 L/hr/kg). For women not receiving oral contraceptives (n = 6) a significant gender-related difference was observed for systemic and oral clearance and for area under the curve and elimination half-life after oral administration. A significant (p < 0.05) reduction in the systemic clearance of midazolam from 28 +/- 9 L/hr to 10 +/- 3 L/hr occurred after clarithromycin administration. Oral midazolam availability was significantly increased from 0.31 +/- 0.1 to 0.75 +/- 0.2 after clarithromycin dosing. Likewise, intestinal and oral availability were significantly increased from 0.42 +/- 0.2 to 0.83 +/- 0.2 and from 0.74 +/- 0.1 to 0.90 +/- 0.04, respectively. A significant correlation was observed between intestinal and oral availability (n = 32, r = 0.98, p < 0.05). After clarithromycin administration, a significant correlation was observed between the initial hepatic or intestinal availability and the relative increase in hepatic or intestinal availability, respectively. Female subjects exhibited a greater extent of interaction after oral and intravenous dosing than male subjects (p < 0.05). CONCLUSION: These data indicate that in addition to the liver, the intestine is a major site of the interaction between oral midazolam and clarithromycin. Interindividual variability in first-pass extraction of high affinity CYP3A substrates such as midazolam is primarily a function of intestinal enzyme activity. PMID- 9728894 TI - Inhibition of chlorzoxazone metabolism, a clinical probe for CYP2E1, by a single ingestion of watercress. AB - To investigate the effect of watercress on the metabolism of chlorzoxazone, an in vivo probe for CYP2E1, the oral pharmacokinetics of chlorzoxazone was studied in 10 healthy volunteers before and after a single ingestion of a watercress homogenate (50 gm). A third chlorzoxazone pharmacokinetic study was performed after a 1-week treatment with isoniazid (300 mg/day), a well-known CYP2E1 inhibitor. Ingestion of watercress or isoniazid did not affect the oral absorption of chlorzoxazone. The area under the chlorzoxazone plasma concentration-time curve was significantly increased by 56% (p < 0.05) after watercress ingestion and by 135% (p < 0.001) with isoniazid treatment. Similarly, chlorzoxazone elimination half-life was prolonged after watercress (53%; p < 0.05) and isoniazid (104%; p < 0.01) administration. These results show that a single ingestion of watercress inhibits the hydroxylation of chlorzoxazone, an in vivo probe for CYP2E1. PMID- 9728895 TI - Selective effects of somatostatin analogs on human drug-metabolizing enzymes. AB - Pharmacologic or surgical manipulation with growth hormone secretion or with the physiologic release of somatostatin and growth hormone-releasing hormone affects some rat liver enzymes, especially the sex-differentiated ones. We investigated the effects of two somatostatin analogs on several enzyme functions in six patients with carcinoid syndrome, using codeine as a probe drug. Codeine was given intravenously and its N- and O-demethylation, as well as 6-glucuronidation catalyzed by CYP3A, CYP2D6, and uridine diphosphate-glucuronosyltransferase, respectively, were studied before and during treatment with somatostatins. After 3 days of treatment with somatostatins the partial metabolic clearance of codeine by N-demethylation decreased by 21% to 64% in all patients (mean change, 44%; p < 0.05), and the clearance by O-demethylation was decreased by 20% to 69% in five of the patients (mean change in all patients, 35%; p < 0.05). In contrast, the partial clearance by 6-glucuronidation and the total systemic clearance of codeine were unchanged. Our results may be caused by the inhibition of growth hormone secretion induced by the somatostatins, inasmuch as direct metabolic interactions with these peptide drugs are improbable. The decline in CYP3A4 and CYP2D6 activity might have clinical implications when substrates of these enzymes with low therapeutic indices are combined with somatostatin analogs. Because the formation of morphine from codeine was altered, the analgesic effect of this drug may be reduced during concomitant treatment with somatostatins. PMID- 9728896 TI - Artemisinin induces omeprazole metabolism in human beings. AB - OBJECTIVE: This study investigated whether time-dependent artemisinin pharmacokinetics correlated to CYP3A4 or CYP2C19 activity in vivo. METHODS: Artemisinin (two oral doses per day of 250 mg) was given to nine healthy Vietnamese subjects for 7 days (day 1 to day 7). Single 20 mg doses of omeprazole were given orally on day -7, day 1, and day 7. Single doses of artemisinin and omeprazole were given in combination on day 14 after a 6-day washout period. The pharmacokinetics of artemisinin, omeprazole, hydroxyomeprazole, and omeprazole sulfone were evaluated on days -7, 1, 7, and 14. On the same days urine was collected for the determination of 6beta-hydroxycortisol and cortisol excretion. RESULTS: Areas under plasma concentration-time curves (AUC) for artemisinin and omeprazole decreased on day 7 to 20% (95% confidence intervals, 13%, 28%) and 35% (25%, 46%), respectively, compared with values on day 1. AUC ratios for hydroxyomeprazole/omeprazole increased 2.2-fold (1.7, 2.7) on day 7 compared with values on day 1. All values were normalized at day 14. There were no significant changes in the omeprazole sulfone/omeprazole ratio or in the 6beta hydroxycortisol/cortisol ratio between the study days. In one subject found to have poor CYP2C19 metabolization, the elimination of omeprazole increased after artemisinin exposure, with no change in the hydroxyomeprazole/omeprazole AUC ratio. CONCLUSION: Artemisinin did not alter CYP3A4 activity, whereas an increase in CYP2C19 activity was observed. The increased elimination of omeprazole in both poor and extensive CYP2C19 metabolizers suggests artemisinin induces both CYP2C19 and another enzyme. PMID- 9728897 TI - Effect of clofibrate on the chiral inversion of ibuprofen in healthy volunteers. AB - OBJECTIVES: To determine the influence of the hypolipidemic drug clofibrate on the stereoselective metabolism of ibuprofen in humans. METHODS: Healthy male subjects (n = 12) ingested a dose of 400 mg pseudoracemic ibuprofen (200 mg R ibuprofen, 160 mg S-ibuprofen, and 40 mg 13C-S-ibuprofen) on two occasions after either pretreatment with clofibrate (2 gm/day over 1 week) or no pretreatment in a randomized order. RESULTS: When subjects were pretreated with clofibrate, clearances of R-ibuprofen and 13C-S-ibuprofen increased significantly from 55.0 and 66.4 ml/min to 186.2 and 106.7 ml/min (p < 0.01), respectively. This increase was similarly reflected in the clearance by inversion of R-ibuprofen (control, 36.0 ml/min; treated, 118.8 ml/min; p < 0.01), as well as in the clearance by noninversion (control, 19.0 ml/min; treated, 67.4 ml/min; p < 0.01). Unbound clearance values significantly increased for R-ibuprofen (control, 19.5 L/min; treated, 38.7 L/min) but not for 13C-S-ibuprofen (11.8 versus 10.6 L/min, respectively). The fractional inversion of ibuprofen calculated from the urinary metabolite data was increased after clofibrate pretreatment (clofibrate group, 66.4%; control, 53.5%; p < 0.01). However, this was not evident when fractional inversion was calculated from the plasma concentration-time data for the unmetabolized drug. CONCLUSIONS: Clofibrate altered the stereoselective disposition of ibuprofen in healthy volunteers by increased formation of R ibuprofenoyl-coenzyme A rather than by an effect on oxidative metabolism of ibuprofen. This interaction has potential therapeutic implications. PMID- 9728899 TI - Concentration-response relationship of levodopa in patients at different stages of Parkinson's disease. AB - OBJECTIVE: To assess differences in the pharmacokinetic and pharmacodynamic relations of levodopa in clinically defined groups and to prove that pharmacokinetic and pharmacodynamic parameters are associated with duration of disease and length of treatment. METHODS: We studied the pharmacokinetic and pharmacodynamic relations of levodopa after a single dose (100 mg levodopa with 25 mg benserazide) among four groups of patients with Parkinson's disease. Group 1 was levodopa-naive patients (n = 8); group 2 was patients in stable condition taking levodopa (n = 10); group 3 was patients with on-and-off fluctuations (n = 11); and group 4 was patients with on-and-off fluctuations and peak-dose dyskinesia (n = 8). The Columbia University Rating Scale was used for clinical assessment. The pharmacokinetic-pharmacodynamic analysis was based on an estimate of the maximal response model with a semiparametric approach to effect-site equilibrium (equilibration half-life). RESULTS: The mean concentration at half maximal effect estimated for the different groups was as follows (mean value +/- SD): group 1, 389 +/- 138 ng x ml(-1); group 2, 346 +/- 203 ng x ml(-1); group 3, 543 +/- 245 ng x ml(-1); group 4, 711 +/- 215 ng x ml(-1). Estimate of the maximal response was determined to be the following: group 1, 10 +/- 3; group 2, 12 +/- 5; group 3, 24 +/- 13; group 4, 18 +/- 7. A significant correlation was observed between duration of Parkinson's disease and mean concentration at half maximal effect (p < 0.001), estimate of maximal response (p < 0.05), and, inversely, equilibration half-life (p < 0.05). CONCLUSIONS: The data suggested that levodopa-naive patients and patients in stable condition taking levodopa do not differ in pharmacokinetic-pharmacodynamic relations, whereas patients with fluctuations, especially patients with peak-dose dyskinesia, exhibit a larger threshold level (mean concentration at half-maximal effect). It was concluded that progression of the disease (loss of endogenous dopamine synthesis and reduced dopamine storage) is reflected by pharmacokinetic and pharmacodynamic parameters that characterize the demand for exogenous dopamine provided by levodopa. PMID- 9728898 TI - Erythromycin and verapamil considerably increase serum simvastatin and simvastatin acid concentrations. AB - OBJECTIVE: To study the effects of erythromycin and verapamil on the pharmacokinetics of simvastatin, an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase. METHODS: A randomized, double-blind crossover study was performed with three phases separated by a washout period of 3 weeks. Twelve young, healthy volunteers took orally either 1.5 gm/day erythromycin, 240 mg/day verapamil, or placebo for 2 days. On day 2, 40 mg simvastatin was administered orally. Serum concentrations of simvastatin, simvastatin acid, erythromycin, verapamil, and norverapamil were measured for up to 24 hours. RESULTS: Erythromycin and verapamil increased mean peak serum concentration (Cmax) of unchanged simvastatin 3.4-fold (p < 0.001) and 2.6-fold (p < 0.05) and the area under the serum simvastatin concentration-time curve from time zero to 24 hours [AUC(0-24)] 6.2-fold (p < 0.001) and 4.6-fold (p < 0.01). Erythromycin increased the mean Cmax of active simvastatin acid fivefold (p < 0.001) and the AUC(0-24) 3.9-fold (p < 0.001). Verapamil increased the Cmax of simvastatin acid 3.4-fold (p < 0.001) and the AUC(0-24) 2.8-fold (p < 0.001). There was more than tenfold interindividual variability in the extent of simvastatin interaction with both erythromycin and verapamil. CONCLUSIONS: Both erythromycin and verapamil interact considerably with simvastatin, probably by inhibiting its cytochrome P450 (CYP) 3A4-mediated metabolism. Concomitant administration of erythromycin, verapamil, or other potent inhibitors of CYP3A4 with simvastatin should be avoided. As an alternative, the dosage of simvastatin should be reduced considerably, that is, by about 50% to 80%, at least when a simvastatin dosage higher than 20 mg/day is used. Possible adverse effects, such as elevation of creatine kinase level and muscle tenderness, should be closely monitored when such combinations are used. PMID- 9728900 TI - Pharmacokinetic-pharmacodynamic modeling of the effects of ivabradine, a direct sinus node inhibitor, on heart rate in healthy volunteers. AB - OBJECTIVE: Ivabradine (S-16257) is a new bradycardic agent with a direct effect on the sinus node. Its N-dealkylated metabolite, S-18982, has shown a bradycardic activity in animals. The aim of this trial was to study the correlation between drug bradycardic activity and plasma levels of the parent compound and its metabolite in healthy volunteers. METHODS: Eighteen healthy volunteers participated in three successive study periods: an oral double-blind period with two parallel groups of doses (10 or 20 mg, single and repeated); a 10 mg intravenous bolus open period; and a final control period. The effects of ivabradine on heart rate were studied at rest and during bicycle exercise tests (at 85% of maximum workload) during 24-hour postdosing, and ivabradine and S 18982 plasma levels were determined simultaneously. RESULTS: The maximal reductions of exercise heart rate were 11% +/- 4% (10 mg) and 18% +/- 6% (20 mg) after single oral doses (p < 0.05) and 18% +/- 4% (10 mg) and 27% +/- 6% (20 mg) after repeated doses (p < 0.01). Maximum heart rate reduction after the intravenous bolus was 19% +/- 4%. After oral administrations an indirect relationship between the bradycardic effect and the plasma concentrations of the two compounds was found. A pharmacokinetic/pharmacodynamic population analysis done with the NONMEM computer program showed that S-18982 contributes in part to the overall activity of ivabradine: modeling suggested that the metabolite is responsible for the initial bradycardic effect, whereas the parent compound is responsible for the duration of action. CONCLUSION: This study shows that ivabradine exerts a dose-dependent bradycardic effect and that its N-dealkylated metabolite contributes to this bradycardic effect. PMID- 9728901 TI - The use of two estrogen preparations (a combined contraceptive pill versus conjugated estrogen cream) intravaginally to treat urogenital symptoms in postmenopausal Thai women: a comparative study. AB - OBJECTIVE: To determine whether the combined contraceptive pill used intravaginally was as effective as the standard conjugated estrogen cream for the treatment of urogenital symptoms in postmenopausal Thai women. SUBJECTS AND METHODS: In a randomized clinical trial, 40 postmenopausal women with urogenital symptoms were randomly allocated to two treatment groups for 8 weeks. The first group (n = 20) received a combined contraceptive pill by the vaginal route, one tablet per week at bedtime for 8 weeks. Each tablet contained 250 microg levonorgestrel plus 30 microg ethinyl estradiol. The second group (n = 20) was given 1 gm of an intravaginal conjugated estrogen cream at bedtime, three times in the first week, twice in the second week, and then once a week for the next 6 weeks (1 gram of conjugated estrogen cream contained 0.625 mg conjugated equine estrogens). Subjects were questioned about their urogenital symptoms, and vaginal cytologic smears, vaginal bacterial cultures, and urine cultures were performed before treatment and after 2, 4, and 8 weeks of therapy. RESULTS: The vaginal pH and the proportion of the fecal type bacteria decreased in both groups, with no statistically significant difference between the groups. The karyopyknotic index and the maturation index were improved during treatment in both groups. An increase in the proportion of lactobacilli were recorded in both groups after therapy, with no significant difference between the two groups. No significant changes were observed in urinary bacteria. The therapy (combined contraceptive pill and estrogen cream) had a marked effect on urogenital symptoms (vaginal dryness, dyspareunia, urinary frequency, and urinary urgency), with impressive improvement comparably in both groups. CONCLUSIONS: A combined contraceptive tablet administered vaginally once a week can alleviate urogenital symptoms in Thai postmenopausal women as effectively as the vaginal estrogen cream. However, the pills are much less expensive and are easily obtained in developing countries. PMID- 9728902 TI - Cognitive performance in elderly subjects after a single dose of befloxatone, a new reversible selective monoamine oxidase A inhibitor. AB - BACKGROUND: Patients with depression often have cognitive and psychomotor performance impairments. Antidepressive treatments can correct these deficits, provided sedative and anticholinergic adverse effects do not add to the preexisting condition, particularly in elderly patients. Newly developed antidepressants therefore should be without deleterious effects on cognitive functions, including memory. Befloxatone is a new antidepressant with a potent, selective, competitive, and reversible inhibitory activity on the A isoform of monoamine oxidase (MAO-A). METHODS: The effects on cognition and psychomotor performance of single oral doses of befloxatone (10 mg) and amitriptyline (50 mg) were compared in a randomized, double-blind, placebo-controlled, three-way crossover design trial in 12 healthy elderly (65 to 85 years) volunteers. The performances of the subjects were evaluated by a comprehensive battery of validated psychometric tests that explored alertness, psychomotor performance, information processing, and memory. Subjective feelings on mood and sleep were rated on visual analog scales. MAO-A inhibition was estimated by multiple titrations of 3,4-dihydrophenylglycol (DHPG) in plasma. RESULTS: Amitriptyline displayed the expected deleterious effects on performance tasks, critical flicker fusion threshold, digit symbol substitution, and body sway, and it deteriorated memory (immediate and delayed free recall of words). In contrast, befloxatone did not impair cognition or psychomotor performance but instead significantly improved the delayed free recall. Amitriptyline adversely affected subjective feelings of alertness and contentedness, but befloxatone permitted sustained alertness and did not alter other subjective feelings or sleep. Concurrently, a single dose of 10 mg befloxatone markedly decreased the DHPG concentration in plasma. CONCLUSION: Contrary to tricyclic antidepressants, whose deleterious effects are greater in elderly subjects, this study demonstrated the safety of befloxatone on cognition and psychomotor performance in elderly subjects. PMID- 9728903 TI - Neuropharmacologic treatment of bronchial asthma with the antidepressant tianeptine: a double-blind, crossover placebo-controlled study. AB - Studies have shown the levels of free serotonin in plasma are increased in symptomatic patients with asthma. In addition, the concentration of free serotonin in symptomatic children with asthma correlates positively with clinical status and negatively with pulmonary function (forced expiratory volume in 1 second [FEV1]). Thus, reducing the concentration of free serotonin in plasma may be useful in treating children with asthma. We studied the effectiveness of tianeptine in treating these patients. Tianeptine is the only drug known to be able to reduce the level of free serotonin in plasma and to enhance the uptake by platelets. Sixty-nine of the 82 children with asthma initially enrolled participated in this study. Children were randomized to receive tianeptine or placebo or both in a double-blind crossover trial. The trial lasted 52 weeks. Tianeptine provoked a dramatic and sudden decrease of both clinical rating and free serotonin plasma levels and an increase in pulmonary function. PMID- 9728904 TI - Interaction between warfarin and 5-fluorouracil, not between warfarin and levamisole. PMID- 9728905 TI - Inducible and reversible gene expression with the rtTA system for the study of memory. AB - To obtain rapidly inducible and reversible expression of transgenes in the forebrain of the mouse, we have combined the reverse tetracycline-controlled transactivator (rtTA) system with the CaMKIIalpha promoter. We show that doxycycline induces maximal gene expression in neurons of the forebrain within 6 days and that this expression can be reversed by removal of doxycycline. Using calcineurin as a test transgene, we show that doxycycline-induced expression impairs both an intermediate form of LTP (I-LTP) in the hippocampus and the storage of spatial memory. The reversibility of the rtTA system in turn allowed us to examine the effects of the transgene on memory retrieval after normal storage had occurred. This examination suggests that retrieval requires some of the same molecular components required for storage. PMID- 9728906 TI - NSF and AMPA receptors get physical. PMID- 9728907 TI - They're playing our song: gene expression and birdsong perception. PMID- 9728908 TI - Brain and language: a perspective from sign language. PMID- 9728909 TI - Neural systems affected in developmental dyslexia revealed by functional neuroimaging. PMID- 9728910 TI - Immunophilins in the nervous system. PMID- 9728911 TI - Separate progenitors for radial and tangential cell dispersion during development of the cerebral neocortex. AB - Cell lineage analyses suggest that cortical neuroblasts are capable of undertaking both radial and tangential modes of cell movement. However, it is unclear whether distinct progenitors are committed to generating neuroblasts that disperse exclusively in either radial or tangential directions. Using highly unbalanced mouse stem cell chimeras, we have identified certain progenitors that are committed to one mode of cell dispersion only. Radially dispersed neurons expressed glutamate, the neurochemical signature of excitatory pyramidal cells. In contrast, tangential progenitors gave rise to widely scattered neurons that are predominantly GABAergic. These results suggest lineage-based mechanisms for early specification of certain progenitors to distinct dispersion pathways and neuronal phenotypes. PMID- 9728912 TI - BDNF regulates reelin expression and Cajal-Retzius cell development in the cerebral cortex. AB - Cajal-Retzius (CR) cells of the cerebral cortex express receptors for the neurotrophin brain-derived neurotrophic factor (BDNF) and downregulate expression of the extracellular matrix protein Reelin during early postnatal development, coincident with the onset of cortical BDNF expression. During this period, mice lacking BDNF have elevated levels of Reelin in CR cells. Acute BDNF stimulation of cortical neuron cultures and overexpression of BDNF in the developing brain of transgenic mice prior to the onset of endogenous production causes a profound, dose-dependent reduction of Reelin expression in CR cells. In addition, overexpression of BDNF produces gaps and heterotopias in the marginal zone and disorganization and aggregation of cortical CR cells and induces several other malformations, including aberrant cortical lamination, similar to the phenotype of reeler mutant mice, which lack Reelin. These results demonstrate a role for BDNF on cortical CR cells and identify Reelin as a direct effector of this neurotrophin during brain development. PMID- 9728913 TI - GFR alpha1-deficient mice have deficits in the enteric nervous system and kidneys. AB - Glial cell line-derived neurotrophic factor (GDNF) signals through a receptor complex composed of the Ret tyrosine kinase and a glycosylphosphatidylinositol- (GPI-) anchored cell surface coreceptor, either GDNF family receptor alpha1 (GFR alpha1) or GFR alpha2. To investigate the usage of these coreceptors for GDNF signaling in vivo, gene targeting was used to produce mice lacking the GFR alpha1 coreceptor. GFR alpha1-deficient mice demonstrate absence of enteric neurons and agenesis of the kidney, characteristics that are reminiscent of both GDNF- and Ret-deficient mice. Midbrain dopaminergic and motor neurons in GFR alpha1 null mice were normal. Minimal or no neuronal losses were observed in a number of peripheral ganglia examined, including the superior cervical and nodose, which are severely affected in both Ret- and GDNF-deficient mice. These results suggest that while stringent physiologic pairing exists between GFR alpha1 and GDNF in renal and enteric nervous system development, significant cross-talk between GDNF and other GFR alpha coreceptors must occur in other neuronal populations. PMID- 9728915 TI - Point mutation in trkB causes loss of NT4-dependent neurons without major effects on diverse BDNF responses. AB - Neurotrophins are a family of soluble ligands that promote the survival and differentiation of peripheral and central neurons and regulate synaptic function. The two neurotrophins, brain-derived neurotrophic factor (BDNF) and neurotrophin 4 (NT4), bind and activate a single high-affinity receptor, TrkB. Experiments in cell culture have revealed that an intact Shc adaptor binding site on TrkB and subsequent activation of the Ras/MAPK pathway are important for neuronal survival and neurite outgrowth. To elucidate the intracellular signaling pathways that mediate the diverse effects of BDNF and NT4 in vivo, we have mutated in the mouse germline the Shc binding site in the trkB gene. This trkB(shc) mutation revealed distinctive responses to BDNF and NT4. While nearly all NT4-dependent sensory neurons were lost in trkB(shc/shc) mutant mice, BDNF-dependent neurons were only modestly affected. Activation of MAP kinases and in vitro survival of cultured trkB(shc/shc) neurons were reduced in response to both neurotrophins, with NT4 being less potent than BDNF, suggesting differential activation of TrkB by the two ligands. Moreover, while the Ras/MAPK pathway is required for in vitro differentiation of neuronal cells, trkB(shc/shc) mutant mice do not show any defects in BDNF-dependent differentiation of CNS neurons or in the function of sensory neurons that mediate innocuous touch. PMID- 9728914 TI - Characterization of neurotrophin and Trk receptor functions in developing sensory ganglia: direct NT-3 activation of TrkB neurons in vivo. AB - Spinal sensory ganglia have been shown to contain neuronal subpopulations with different functions and neurotrophin dependencies. Neurotrophins act, in large part, through Trk receptor tyrosine kinases: nerve growth factor (NGF) via TrkA, brain-derived neurotrophic factor (BDNF) and neurotrophin-4/5 (NT-4/5) via TrkB, and neurotrophin-3 (NT-3) via TrkC. In the present paper, we use antibodies to TrkA, TrkB, and TrkC to characterize their expression patterns and to determine which subpopulations of cells are lost in mice lacking individual neurotrophins or Trk receptors. Despite previous reports of Trk receptor mRNAs in neural crest cells, we detect Trk receptor proteins only in neurons and not in neural crest cells or neuronal precursors. Comparisons of neonatal mice deficient in NT-3 or its cognate receptor TrkC have shown that there is a much greater deficiency in spinal sensory neurons in the former, suggesting that NT-3 may activate receptors in addition to TrkC. Using the same antibodies, we show that, during the major period of neurogenesis, NT-3 is required to maintain neurons that express TrkB in addition to those that express TrkC but is not essential for neurons expressing TrkA. Results also indicate that survival of cells expressing both receptors can be maintained by activation of either one alone. NT-3 can thus activate more than one Trk receptor in vivo, which when coexpressed are functionally redundant. PMID- 9728916 TI - Long-term depression at thalamocortical synapses in developing rat somatosensory cortex. AB - Sensory experience during an early critical period guides the development of thalamocortical circuits in many cortical areas. This process has been hypothesized to involve long-term potentiation (LTP) and long-term depression (LTD) at thalamocortical synapses. Here, we show that thalamocortical synapses in rat barrel cortex can express LTD, and that LTD is most readily induced during a developmental period that is similar to the critical period for thalamocortical plasticity in vivo. Thalamocortical LTD is homosynaptic and dependent on activation of N-methyl-D-aspartate (NMDA) receptors. The age-related decline of LTD is not due to changes in inhibition nor to changes in NMDA receptor voltage dependence. Minimal stimulation experiments indicate that, unlike thalamocortical LTP, thalamocortical LTD is not associated with a significant change in failure rate. The existence of LTD and its developmental time course suggest that LTD, like LTP, may contribute to the refinement of thalamocortical inputs in vivo. PMID- 9728917 TI - Toward a song code: evidence for a syllabic representation in the canary brain. AB - We show that presentation of individual canary song syllables results in distinct expression patterns of the immediate-early gene ZENK in the caudomedial neostriatum (NCM) of adult canaries. Information on the spatial distribution and labeling of stained cells provides for a classification of ZENK patterns that (1) accords to the organization of stimuli into families, (2) preserves the stimuli intrafamily relationships, and (3) confers salience to natural over artificial stimuli, resulting in a nonclassical tonotopic map. Moreover, complex syllable maps cannot be reduced to any linear combinations of simple syllable maps. These properties arise from the collective response of NCM neurons to auditory stimuli, rather than from the behavior of single neurons. The syllabic representation described here may constitute an important step toward deciphering the rules of birdsong auditory representation. PMID- 9728918 TI - An area within human ventral cortex sensitive to "building" stimuli: evidence and implications. AB - Isolated, ventral brain lesions in humans occasionally produce specific impairments in the ability to use landmarks, particularly buildings, for way finding. Using functional MRI, we tested the hypothesis that there exists a cortical region specialized for the perception of buildings. Across subjects, a region straddling the right lingual sulcus was identified that possessed the functional correlates predicted for a specialized building area. A series of experiments discounted several alternative explanations for the behavior of this site. These results are discussed in terms of their impact upon our understanding of the functional structure of visual processing, disorders of topographical disorientation, and the influence of environmental conditions upon neural organization. PMID- 9728920 TI - Interaction of the N-ethylmaleimide-sensitive factor with AMPA receptors. AB - Glutamate receptors mediate the majority of rapid excitatory synaptic transmission in the central nervous system (CNS) and play important roles in synaptic plasticity and neuronal development. Recently, protein-protein interactions with the C-terminal domain of glutamate receptor subunits have been shown to be involved in the modulation of receptor function and clustering at excitatory synapses. In this paper, we have found that the N-ethylmaleimide sensitive factor (NSF), a protein involved in membrane fusion events, specifically interacts with the C terminus of the GluR2 and GluR4c subunits of AMPA receptors in vitro and in vivo. Moreover, intracellular perfusion of neurons with a synthetic peptide that competes with the interaction of NSF and AMPA receptor subunits rapidly decreases the amplitude of miniature excitatory postsynaptic currents (mEPSCs), suggesting that NSF regulates AMPA receptor function. PMID- 9728919 TI - UNC-115, a conserved protein with predicted LIM and actin-binding domains, mediates axon guidance in C. elegans. AB - Axon guidance receptors modulate the growth cone cytoskeleton through signaling pathways that are not well understood. Here, we describe the C. elegans unc-115 gene, which encodes a candidate cytoskeletal linker protein that acts in axon guidance. unc-115 mutants have defects in a subset of axons, particularly as the affected axons change environments during outgrowth. The unc-115 gene encodes a putative actin-binding protein that is similar to the human actin-binding protein abLIM/limatin; it has a villin headpiece domain and three LIM domains that could mediate protein interactions. unc-115 is expressed in neurons during their development and is required cell-autonomously in certain neurons for normal axon guidance. We propose that UNC-115 modulates the growth cone actin cytoskeleton in response to signals received by growth cone receptors. PMID- 9728921 TI - Temperature-sensitive paralytic mutations demonstrate that synaptic exocytosis requires SNARE complex assembly and disassembly. AB - The neuronal SNARE complex is formed via the interaction of synaptobrevin with syntaxin and SNAP-25. Purified SNARE proteins assemble spontaneously, while disassembly requires the ATPase NSF. Cycles of assembly and disassembly have been proposed to drive lipid bilayer fusion. However, this hypothesis remains to be tested in vivo. We have isolated a Drosophila temperature-sensitive paralytic mutation in syntaxin that rapidly blocks synaptic transmission at nonpermissive temperatures. This paralytic mutation specifically and selectively decreases binding to synaptobrevin and abolishes assembly of the 7S SNARE complex. Temperature-sensitive paralytic mutations in NSF (comatose) also block synaptic transmission, but over a much slower time course and with the accumulation of syntaxin and SNARE complexes on synaptic vesicles. These results provide in vivo evidence that cycles of assembly and disassembly of SNARE complexes drive membrane trafficking at synapses. PMID- 9728922 TI - Activity-dependent modulation of the rate at which synaptic vesicles become available to undergo exocytosis. AB - The number of vesicles contained in the readily releasable pool at excitatory hippocampal synapses has recently been identified as a major determinant of the strength of these synapses. Here, we show that the rate at which this pool refills following depletion is variable from neuron to neuron and can be increased by the accumulation of intracellular calcium during action potential mediated activation of the synapses. The refilling rate nearly doubles during the first second of a high frequency train of action potentials and does not increase further with additional stimulation. During periods of rest, the rate relaxes back to its original value, with a time constant of about 10 s. Since this refilling rate helps set the strength of synapses during high frequency bursts of action potentials and is modulated by physiological signaling, it is an attractive candidate point of control in the storage and manipulation of information by the central nervous system. PMID- 9728923 TI - Glutamate release monitored with astrocyte transporter currents during LTP. AB - Long-term potentiation (LTP) of synaptic transmission in the CA1 region of the hippocampus is thought to result from either increased transmitter release, heightened postsynaptic sensitivity, or a combination of the two. We have measured evoked glutamate release from Schaffer collateral/commissural fiber terminals in CA1 by recording synaptically activated glutamate transporter currents in hippocampal astrocytes located in stratum radiatum. Although several manipulations of release probability caused parallel changes in extracellular field potentials and synaptically activated transporter current amplitudes, induction of LTP failed to alter transporter-mediated responses, suggesting that LTP does not alter the amount of glutamate released upon synaptic stimulation. PMID- 9728924 TI - Monitoring glutamate release during LTP with glial transporter currents. AB - Although much has been learned about the mechanisms underlying NMDA receptor dependent long-term potentiation (LTP), considerable debate remains as to whether LTP is expressed as an increase in the synaptic release of glutamate or as an increase in the sensitivity of the postsynaptic glutamate receptors. We have directly measured changes in the synaptic release of glutamate by recording synaptically evoked glial glutamate transporter currents with whole-cell recording. Glial cell responses were very sensitive to manipulations known to change the release of glutamate yet remained constant during LTP. These results argue strongly for a postsynaptic expression mechanism for LTP. PMID- 9728925 TI - Calmodulin mediates calcium-dependent inactivation of N-methyl-D-aspartate receptors. AB - Ca2+ influx through N-methyl-D-aspartate (NMDA) receptors activates signal transduction pathways critical for many forms of synaptic plasticity in the brain. NMDA receptor-mediated Ca2+ influx also downregulates the gating of NMDA channels through a process called Ca2+-dependent inactivation (CDI). Recent studies have demonstrated that the calcium binding protein calmodulin directly interacts with NMDA receptors, suggesting that calmodulin may play a role in CDI. We report here that the mutation of a specific calmodulin binding site in the CO region of the NR1 subunit of the NMDA receptor blocks CDI. Moreover, intracellular infusion of a calmodulin inhibitory peptide markedly reduces CDI of both recombinant and neuronal NMDA receptors. Furthermore, this inactivating effect of calmodulin can be prevented by coexpressing a region of the cytoskeletal protein alpha-actinin2 known to interact with the CO region of NR1. Taken together, these results demonstrate that the binding of Ca2+/calmodulin to NR1 mediates CDI of the NMDA receptor and suggest that inactivation occurs via Ca2+/calmodulin-dependent release of the receptor complex from the neuronal cytoskeleton. PMID- 9728926 TI - Synthesized allosteric effectors of the hemoglobin molecule: a possible mechanism for improved erythrocyte oxygen release capability in hemoglobinopathy H disease. AB - Patients with the nondeletion genotype of hemoglobinopathy H (HbH or beta4) disease have higher proportions of HbH and more severe tissue hypoxia than patients with the deletion genotype. Because these patients' red blood cells (RBCs) contain mainly two Hb species, HbH and HbA, the high proportion of HbA can be exploited by lowering its oxygen affinity; this would probably increase oxygen delivery to the RBCs and improve the patients' clinical phenotype. Allosteric effectors that induce a low-affinity Hb may be useful in this regard. We investigated the effect of a bezafibrate derivative, RSR-4, on the oxygen affinity of RBCs and purified hemolysates containing HbA and HbH. This allosteric effector crosses RBC membranes and binds reversibly to the alpha-chains of deoxy Hb, decreasing hemoglobin oxygen affinity. The blood used was obtained from a patient with HbH disease (alphaTSaudi homozygote) whose HbH level was 33.5% as measured by high-performance liquid chromatography. Oxygen binding studies were performed in RBCs and purified hemolysates. RBCs incubated in the presence of 500 microM RSR-4 (2-[[[(3,5-dichloroanilino)-carbonyl]methyl]phenoxy]-2-methylpropi onic acid) in standard conditions (pH 7.4, 0.14 M NaCl, 37 degrees C) displayed an increase in their P50 value from 14.5 to 35.2 mm Hg. Oxygen binding studies in purified stripped hemolysates (pH 7.2, 0.1 M NaCl, 25 degrees C) showed that addition of both 500 microM RSR-4 and 1 mM of 2,3 diphosphoglycerate (DPG) led to an 11-fold decrease in oxygen affinity, whereas the addition of the natural effector DPG or RSR-4 alone produced a 2.7- and 5.7-fold decrease, respectively. In both cases, the oxygen equilibrium curves (OECs) were biphasic due to the presence of the noncooperative, high-oxygen-affinity HbH (beta4) component. After addition of RSR-4, the lower part of the OEC (corresponding to HbH) was not shifted compared with the upper part (corresponding to HbA). These results were confirmed by kinetic studies of CO recombination. Both experiments demonstrated that RSR-4 does not affect beta4 Hb. Our findings provide an experimental model for lowering the oxygen affinity of HbA in HbH-containing cells and suggest that the oxygen delivery capability of the latter would be thereby improved. PMID- 9728927 TI - A partial conditioning strategy for achieving mixed chimerism in the rat: tacrolimus and anti-lymphocyte serum substantially reduce the minimum radiation dose for engraftment. AB - Development of partial conditioning strategies to achieve reliable engraftment of allogeneic bone marrow with minimum recipient morbidity could extend the therapeutic application of bone marrow transplantation (BMT) to enzyme deficiency states, hemoglobinopathies, autoimmune diseases, and the induction of tolerance for solid organ and cellular allografts. In this study we describe a nonmyeloablative rat BMT model and examine the effect of clinically available immunosuppressants on the minimum amount of total body irradiation (TBI) required for allogeneic engraftment. Donor ACI marrow was depleted of T cells using immunomagnetic beads and transplanted to major histocompatibility complex- and minor antigen-mismatched Wistar Furth (WF) rats (ACI --> WF) conditioned with varying doses of TBI. Recipients conditioned with TBI alone required myeloablation with 1000 cGy for reliable allogeneic marrow engraftment. Administration to WF recipients of a single dose of anti-lymphocyte serum (ALS) 5 days prior to BMT together with a limited course of tacrolimus (1 mg/kg/day) resulted in engraftment of ACI bone marrow at only 500 cGy TBI. ACI --> WF recipients were stable mixed chimeras (mean donor chimerism 49% at 330 days post BMT). Chimerism was multilineage. All recipient animals were free of graft-versus host disease. These results suggest that a nonmyeloablative conditioning strategy based on low-dose TBI and a limited course of tacrolimus plus ALS can produce long-term mixed multilineage chimerism. PMID- 9728928 TI - Digital image analysis of hematopoietic colonies in vitro. AB - A system for automatic analysis of in vitro hematopoietic colonies is described and evaluated. With the standard resolution provided by video cameras, the improvement in visualization obtained using features other than size and darkness when classifying potential colonies appears to be limited. We confirmed this by comparing results obtained with the test system with those obtained with a commercial one. However, for some applications it may be useful to supplement the system with specific methods, e.g., to separate merged colonies. Digital image analyses provide new possibilities, for instance of measuring the total cellularity of the dish or analyzing colonies according to the size and cell density of each colony. Examples provided are time course studies of colony development, cellularity feedback effects on colony sizes, and bell-shaped dose response curves for the growth stimulation obtained by certain conditioned media on a subpopulation of progenitor cells that gives rise to large colonies. PMID- 9728929 TI - Delayed neutrophil apoptosis after total body irradiation in mice. The role of granulocyte-macrophage colony-stimulating factor in neutrophil function. AB - We performed an in vitro study of the long-term effects of a sublethal dose (5 Gy) of x-irradiation on the survival and function of neutrophils in adult mice. For this purpose, we incubated control neutrophils harvested from long-term bone marrow cultures (LTBMCs) with supernatant withdrawn from cultures obtained in adult mice 6 or 9 months postirradiation. We noted a significant increase in superoxide anion production, NADPH, and protein levels in these cells after 3, 6, and 15 hours of incubation compared with the same cells incubated with supernatant from control LTBMCs. We also observed a delay in apoptosis that was correlated with maintenance of adenosine triphosphate levels and survival. Similar differences were found when control LTBMC neutrophils were incubated with granulocyte-macrophage colony-stimulating factor (GM-CSF) (1.3 nM). Indeed, enzyme-linked immunosorbent assay revealed a significant overproduction of this cytokine, together with higher interleukin (IL)-6 and IL-3 levels, in the supernatant from cultures of irradiated mice. Our results suggest that GM-CSF is one of the cytokines responsible for promoting the survival and activation of neutrophil function after total body irradiation. PMID- 9728930 TI - Bone marrow repopulation by human marrow stem cells after long-term expansion culture on a porcine endothelial cell line. AB - In vitro exposure of murine hematopoietic stem cells (HSCs) to cell cycle inducing cytokines has been shown to result in a defect in the ability of these cells to engraft. We used a porcine microvascular endothelial cell (PMVEC) line in conjunction with exogenous interleukin (IL)-3, IL-6, granulocyte-macrophage colony-stimulating factor (GM-CSF), and stem cell factor (SCF) to expand human HSCs that express the CD34 and Thy-1 antigens but lack lineage-associated markers (CD34+Thy-1+Lin- cells). Ex vivo expansion of hematopoietic cells was evaluated in comparison to stromal cell-free, cytokine-supplemented cultures. Cells expressing the CD34+Thy-1+Lin- phenotype were detectable in both culture systems for up to 3 weeks. These cells were reisolated from the cultures and their ability to engraft human fetal bones implanted into SCID mice (SCID-hu bone) was tested. HSCs expanded in PMVEC coculture were consistently capable of competitive marrow repopulation with multilineage (CD19+ B lymphoid, CD33+ myeloid, and CD34+ cells) progeny present 8 weeks postengraftment. In contrast, grafts composed of cells expanded in stroma-free cultures did not lead to multilineage SCID-hu bone repopulation. Proliferation analysis revealed that by 1 week of culture more than 80% of the cells in the PMVEC cocultures expressing the primitive CD34+CD38- phenotype had undergone cell division. Fewer than 1% of the cells that proliferated in the absence of stromal cells remained CD34+CD38-. These data suggest that the proliferation of HSCs in the presence of IL-3, IL-6, GM-CSF, and SCF without stromal cell support may result in impairment of engraftment capacity, which may be overcome by coculture with PMVECs. PMID- 9728931 TI - Comparison of amphotropic and pseudotyped VSV-G retroviral transduction in human CD34+ peripheral blood progenitor cells from adult donors with HIV-1 infection or cancer. AB - In this study we compared the transduction efficiency of conventional amphotropic MoMLV (LPONL[A]) with the MoMLV pseudotyped with that of VSV-G (LPONL[G]) in peripheral blood progenitor cells (PBPCs) from cancer patients and human immunodeficiency virus (HIV)-infected donors. The results showed that LPONL(A) and LPONL(G) infected the progenitor cells from these sources with equal efficiencies. The transgene neoR was detectable by polymerase chain reaction assay in colonies from 14-day colony-forming unit (CFU) assays and in those derived from long-term culture-initiating cell (LTC-ICs) assays. Although the overall levels of transduction efficiency were similar in cord blood and PBPCs from noninfected cancer donors (25-22%) when either LPONL(G) or LPONL(A) was used, they were significantly lower in HIV-1-infected donors compared with noninfected cancer donors when LPONL(G) was used (13 vs. 25%; p = 0.027), and when LPONL(A) was used (12 vs. 22%; p = 0.087). The clonogenic potentials of infected and noninfected CD34+ cells were similar; thus no toxicity could be attributed to the virus preparation. We conclude that PBPCs from HIV-1-infected individuals are transduced less efficiently than those from non-HIV-infected cancer donors. Nonetheless, PBPCs from HIV-infected persons serve as potential targets in gene therapy for acquired immune deficiency syndrome. PMID- 9728932 TI - Leukapheresis products in multiple myeloma: lower tumor load after mobilization with cyclophosphamide plus granulocyte colony-stimulating factor (G-CSF) compared with G-CSF alone. AB - High-dose therapy with autografting of peripheral blood stem cells (PBSCs) has become an accepted treatment modality. However, gene-marking studies in patients with acute myeloid leukemia and neuroblastoma have revealed that malignant cells reinfused along with leukapheresis products (LPs) contribute to relapse. Thus, a reduction in the number of malignant cells in autografts is desirable. We analyzed the percentage of malignant cells and the number of CD34+ PBSCs in LPs mobilized by granulocyte colony-stimulating factor (G-CSF) alone (LP-S) compared with high-dose cyclophosphamide plus G-CSF (LP-CY) in patients with multiple myeloma (MM). A quantitative polymerase chain reaction assay involving CDR3 specific primers based on the method of limiting dilutions was used to determine the tumor loads of LPs. Sixteen LPs from eight patients with MM were analyzed intraindividually in matched pairs. The percentage of malignant cells was lower in LP-CY (p = 0.017; median 0.0067 vs. 0.009%), whereas the number of CD34+ cells was higher (p = 0.012; median 0.3 vs. 0.095%). The calculated number of malignant cells per CD34+ cell was significantly lower in LP-CY as well (p = 0.017). We conclude that mobilization by cyclophosphamide plus G-CSF leads to a lower number of malignant cells per CD34+ cell in LPs compared with G-CSF alone. PMID- 9728933 TI - Localization to chromosome 11 of a gene encoding a human minor histocompatibility antigen. AB - The graft-versus-host disease (GVHD) seen in human leukocyte antigen (HLA) matched sibling bone marrow transplants is by definition due to the "minor" histocompatibility antigens (mHAs) encoded outside the HLA region of human chromosome 6. Few of these antigens have been characterized in humans, and in general the locations of the encoding loci are unknown. Genetic experiments performed in mice have identified many mHAs, but only a few genes have been identified. Using T lymphocyte clones reactive with specific mHAs, combined with genetic linkage analysis, we identified two distinct loci in a single patient, each locus encoding an antigen presented to a T cell clone by HLA-B7. The technique used in this study should allow a rough enumeration of the number of mHAs in humans that are capable of eliciting T cell responses in vivo. Whether these T cell responses correlate with clinical GVHD is not yet clear. PMID- 9728934 TI - Effect of a bcl-2 transgene on production and localization of precursor B cells in mouse bone marrow. AB - Many B cell precursors die while differentiating in mouse bone marrow. To ascertain the mechanisms involved in this process, populations of B lineage cells and their tissue localization were analyzed in bone marrow of transgenic mice overexpressing the apoptosis inhibitor, Bcl-2. Immunofluorescence labeling and mitotic arrest were used to quantitate the number and proliferative activity of mu- pro-B cells (terminal deoxynucleotidyl transferase [TdT]+B220-, TdT+B220+, and TdT-B220+); pre-B cells (cmu+); and B cells (smu+). Mature B cells (IgM+IgD+) were increased 16- to 20-fold. In addition, immature B lymphocytes (IgM+IgD /low), representing newly formed cells, were increased three- to sixfold, whereas pre-B cells and late pro-B cells were increased 30 to 60% in production rate. Earlier pro-B cells expressing TdT were unaffected. In spleen, both mature and immature B cells were greatly increased, but cells of precursor phenotype were few and TdT+ cells were absent. The in vivo location of B cells was examined by autoradiography using light and electron microscopy after intravenous injection of 125I-labeled antibodies. B lineage cells (B220+) were increased throughout bone marrow, often within dilated venous sinusoids, particularly in subosteal regions. Many intravascular and perisinusoidal cells were IgDhigh mature B lymphocytes. In contrast, many other IgM+ and IgDlow immature B lymphocytes clustered extravascularly around the central venous sinus. Plasma cells with distended endoplasmic reticulum were numerous. These findings provide evidence that, in addition to expanding the recirculating pool of B cells entering bone marrow from the blood stream, high levels of Bcl-2 can inhibit some of the apoptosis occurring during B cell differentiation, thereby expanding populations of B lymphopoietic precursor cells within the bone marrow parenchyma. PMID- 9728935 TI - T-lymphopoietic capacity of cord blood-derived CD34+ progenitor cells. AB - Umbilical cord blood contains an abundance of CD34+ hematopoietic progenitor cells and has been used in transplantation as an alternative to adult bone marrow or mobilized peripheral blood. Although efficient myelomonocytic, erythroid, and B lymphoid differentiation has been shown in highly purified cord blood CD34+ mononuclear cells lacking expression of lineage-specific antigens, generation of functional T cells has not been previously documented. Exploiting two recently developed, complementary thymic stromal monolayer systems, we show here that immature hematopoietic progenitor cells (CD34+lineage-) from human and rhesus monkey cord blood mononuclear cells undergo T lymphopoiesis in a manner that recapitulates T cell ontogeny in vivo. After 2 weeks of proliferation, cultures contained myeloid [corrected] cells and discrete populations of CD4+CD8+ (double positive) immature T lymphocytes, followed after an additional 2 weeks by the appearance of single-positive CD4+CD8- and CD4-CD8+ T cells that coexpressed CD3. The T lymphoid phenotype was confirmed at the transcriptional level by the presence of the lymphoid-restricted genes RAG-2 and T cell receptor. T cells generated from cord blood progenitors in these systems exhibited immunofunction as assessed by alloreactive responses in mixed lymphocyte reactions. These findings were comparable between human and rhesus progenitor cells and closely resemble previous data using adult bone marrow CD34+ cells in these models. Together, these observations demonstrate that cord blood contains abundant lymphoid progenitors that undergo T lymphopoiesis in vitro, suggesting the full multipotentiality of this stem cell source and its validity in investigating T lymphoid differentiation. PMID- 9728936 TI - Analysis of cell death in myeloid cells inducibly expressing the cell cycle protein p55Cdc. AB - p55Cdc, a cell cycle protein is expressed in cycling mammalian cells and is required for normal cell division. Expression of this protein is regulated during the cell cycle, peaking in late G1 and S. We have previously shown that constitutive expression of p55Cdc results in inhibition of granulocyte differentiation. Degradation of p55Cdc is also required for apoptosis in growth factor and serum starved cells. In the present study we prepared stably transfected cells conditionally expressing p55Cdc in response to zinc stimulation to investigate the role of inducible p55Cdc expression in apoptosis of myeloid cells. We report that inducible expression of p55Cdc in the myeloid leukemic cell line 32Dc13 resulted in increased cell death. p55Cdc overexpression led to a statistically significant decrease in the viability of 32Dc13 cells compared with that of control cells. Furthermore, cell staining and flow cytometry analysis revealed that p55Cdc-overexpressing 32Dc13 cells progressed to apoptosis much earlier than uninduced cells. These results suggest that inducible expression of p55Cdc leads to earlier increases in cell death in the absence of growth factor and serum in myeloid leukemic cells. PMID- 9728937 TI - Treatment of postural hypotension. PMID- 9728938 TI - Corticosteroids and multiple sclerosis. PMID- 9728939 TI - Neurology and the gastrointestinal system. AB - Both achalasia and Hirchsprung's disease arise from defects of innervation of the oesophagus and distal large bowel respectively. Their consequences are confined to disorders of motility in the relevant part of the gastrointestinal tract. Many neurogenic and primary muscle disorders are associated with abnormalities of gut motility. Stroke, even when unilateral, is commonly associated with dysphagia. Transcranial magnetoelectric stimulation has established that the pharyngeal phase of swallowing tends to receive its innervation principally from one hemisphere. In many neurological disorders, dysphagia is only one part of the clinical picture but in some--for example, the Chiari malformation--dysphagia may be the sole or major feature. Disturbances of small and large bowel motility, when seen in neurogenic disorders, are associated with autonomic neuropathy and are particularly common in diabetes mellitus. Primary muscle disorders can lead to dysphagia (for example, with polymyositis or oculopharyngeal dystrophy) or defects of large bowel motility (for example, with Duchenne's muscular dystrophy). Primary gut disorders particularly associated with neurological disease include pernicious anaemia, nicotinamide and thiamine deficiencies, selective vitamin E deficiency, and coeliac disease. Inflammatory bowel disease is associated with thromboembolic complications which may include the CNS, inflammatory muscle disease, and abnormalities on MRI of the brain of uncertain relevance. Whipple's disease is a rare condition which sometimes is largely or entirely confined to the CNS. In such cases, a particular neurological presentation can indicate the diagnosis. PMID- 9728940 TI - Evidence of CNS impairment in HIV infection: clinical, neuropsychological, EEG, and MRI/MRS study. AB - OBJECTIVES: To identify by clinical examination, EEG, MRI, and proton spectroscopy, and neuropsychological assessment the prevalence of signs of CNS involvement in patients infected with HIV, and to relate such findings to the evidence of immunosuppression. METHODS: The design was a cross sectional analysis of a cohort of male patients with infected HIV with an AIDS defining diagnosis or low CD4 count (<350), and seropositive asymptomatic subjects, both groups being followed up in a longitudinal study. Control groups consisted of seronegative subjects from the same genitourinary medicine clinics. RESULTS: This report sets out the cross sectional findings at the seventh visit in the longitudinal study. Patients with AIDS had more signs of neurological dysfunction, poorer performance on a neuropsychological test battery, were more likely to have an abnormal EEG, and to have abnormalities on MRI. They more often had cerebral atrophy, abnormal appearing white matter, and abnormal relaxometry and spectroscopy. There was little evidence of abnormality in seropositive people who had a CD4 count >350 compared with seronegative people from a similar background. CONCLUSIONS: Detailed testing failed to disclose significant CNS impairment without immunosuppression in men infected with HIV. Findings from MRI and magnetic resonance spectroscopy (MRS) correlated with those of the neurological examination and neuropsychological assessment. A combination of such assessments offers a simple surrogate for studies of CNS involvement in HIV disease. PMID- 9728941 TI - Dural arteriovenous fistulas as a cause of intracranial hypertension due to impairment of cranial venous outflow. AB - OBJECTIVES: A retrospective study was carried out on 13 patients with intracranial dural arteriovenous fistulas (DAVFs) who presented with isolated or associated signs of intracranial hypertension. METHODS: Nine patients presented with symptoms of intracranial hypertension at the time of diagnosis. Ocular fundoscopy available in 12 patients showed bilateral papilloedema in eight and optic disk atrophy in four. Clinical evolution was particularly noticeable in five patients because of chronic (two patients) or acute (after lumbar shunting or puncture: three patients, one death) tonsillar herniation. RESULTS: Two patients had a type I fistula (drainage into a sinus, with a normal antegrade flow direction). The remaining 11 had type II fistulas (drainage into a sinus, with abnormal retrograde venous drainage into sinuses or cortical veins). Stenosis or thrombosis of the sinus(es) distal to the fistula was present in five patients. The cerebral venous drainage was abnormal in all patients. CONCLUSION: Type II (and some type I) DAVFs may present as isolated intracranial hypertension mimicking benign intracranial hypertension. Normal cerebral angiography should be added as a fifth criterion of benign intracranial hypertension. The cerebral venous drainage pattern must be carefully studied by contralateral carotid and vertebral artery injections to correctly evaluate the impairment of the cerebral venous outflow. Lumbar CSF diversion (puncture or shunting) may induce acute tonsillar herniation and should be avoided absolutely. DAVF may induce intracranial hypertension, which has a poor long term prognosis and may lead to an important loss of visual acuity and chronic tonsillar herniation. Consequently, patients with intracranial hypertension must be treated, even aggressively, to obliterate the fistula or at least to reduce the arterial flow and to restore a normal cerebral venous drainage. The endovascular treatment may associate arterial or transvenous embolisation and/or surgery. Patients in whom the fistula is not obliterated after an endovascular therapeutic procedure, need continuous clinical and angiographical follow up. PMID- 9728942 TI - Cerebral blood flow and metabolism in patients with silent brain infarction: occult misery perfusion in the cerebral cortex. AB - OBJECTIVES: Silent brain infarction (SBI) is of growing interest as a possible risk factor for symptomatic stroke. Although morphological characteristics of SBI have been well defined, their characteristic patterns of cerebral blood flow (CBF) and metabolism are in dispute. The purpose of this study was to elucidate CBF and metabolism in patients with SBI in relation to symptomatic stroke. METHODS: The patients underwent PET and were separated into three groups; control group (C group), with no lesions on CT (n=9, mean age 57), SBI group, with no neurological signs or history of stroke, but with ischaemic lesions on CT (n=9, mean age 63), and brain infarction group (BI group), with neurological deficits and compatible CT lesions in the area supplied by perforating arteries (n=19, mean age 56). Regional CBF, oxygen extraction fraction (OEF), cerebral metabolic rate for oxygen (CMRO2), and cerebral blood volume (CBV) were measured by PET. RESULTS: Mean values for CBF to the cerebral cortex and deep grey matter were lower in the SBI group (31.6 (SD 5.8) and 34.3 (SD 6.9) ml/100 g/min, respectively) and in the BI group (30.8 (SD 5.2), 33.9 (SD 5.9), respectively) than in the C group (36.0 (SD 6.6) and 43.5 (SD 9.5), respectively). Although mean CMRO2 of deep grey matter (2.36 (SD 0.52) ml/100 g/min) was significantly decreased in the SBI group compared with the C group (2.76 (SD 0.480), p<0.01), CMRO2 of the cortical area was as well preserved in the SBI patients (2.36 (SD 0.39)) as in the controls (2.48 (SD 0.32)) with a compensatory increase of mean OEF (0.45 (SD 0.06) and 0.41 (SD 0.05), respectively). CONCLUSIONS: Patients with SBI showed decreased CBF and CMRO2 in deep grey matter. On the other hand, decreased CBF with milder increased OEF, resulting in preserved CMRO2 in the cerebral cortex indicates the presence of occult misery perfusion, suggesting that patients with SBI have reduced cerebral perfusional reserves. PMID- 9728943 TI - Major decrease in the volume of the entorhinal cortex in patients with Alzheimer's disease carrying the apolipoprotein E epsilon4 allele. AB - OBJECTIVE: Recent evidence indicates that the apolipoprotein E (ApoE) epsilon4 allele is a risk factor for developing Alzheimer's disease. It has also been proposed that it is associated with increased counts of amyloid plaques and neurofibrillary tangles that in turn are neuropathological hallmarks initially appearing in the medial temporal lobe structures in Alzheimer's disease. In this study, the effect of the ApoE epsilon4 allele on the volume of the entorhinal cortex was evaluated in vivo. METHODS: The volume of the entorhinal cortex was measured on MR images using a recently designed histology based protocol in 16 patients with Alzheimer's disease with ApoE epsilon4 (mean age 70.4 (SD 9.9)), 11 patients with Alzheimer's disease without ApoE epsilon4 (mean age 69.1 (SD7.1)), and in 31 healthy age and sex matched normal controls (72.2 (SD 3.9)). The patients met the NINCDS-ADRDA criteria for probable Alzheimer's disease and were in mild to moderate stages of the disease. MRI was performed with a 1.5 Tesla Magnetom and a 3D technique permitting the reconstruction of 2.0 mm thick contiguous slices perpendicular to the axis of the anterior-posterior commissure. RESULTS: The patients with Alzheimer's disease without the ApoE epsilon4 allele had atrophy in the entorhinal cortex, the volume was reduced by 27% compared with control subjects. However, the most prominent shrinkage (45%) in the entorhinal cortex was seen in patients with Alzheimer's disease with the ApoE epsilon4 allele (p=0.0001). The effect of epsilon4 on the entorhinal cortex volume was especially prominent in female patients with Alzheimer's disease compared to male patients with Alzheimer's disease (p=0.014). Additionally, patients with the ApoE epsilon4 allele had inferior performance in verbal and visual memory functions than those without the allele CONCLUSIONS: Volumetric MRI measurements disclose that ApoE epsilon4 is associated with the degree of atrophy in the entorhinal cortex in early Alzheimer's disease, this effect being especially prominent in female patients with Alzheimer's disease. PMID- 9728944 TI - Motor switching abilities in Parkinson's disease and old age: temporal aspects. AB - OBJECTIVES: To investigate capabilities of arm trajectory modification in patients with Parkinson's disease and elderly subjects using a double step target displacement paradigm. METHODS: Nine patients with Parkinson's disease and seven age matched control subjects were instructed to move a stylus towards visual targets presented on a digitising table. Within each session, in some trials the target location was changed before initiation of movement and the subjects were to modify their movements towards the new target (switching trials). In other trials the target location was not changed (control trials). This procedure was repeated for four different target configurations, using interstimulus time intervals of six different durations. The subjects' hand trajectories were recorded and their kinematic characteristics were analysed. RESULTS: In switching trials, about 40% of the movements were aimed directly toward the final target location in both groups. When the trajectories were initially directed toward the first target and then modified toward the second, the reaction time (RT) to the second stimulus (RT2) was longer than to the first stimulus (RT1). The RT2/RT1 ratio was significantly larger in patients with Parkinson's disease than in healthy elderly subjects. CONCLUSIONS: Patients with Parkinson's disease and elderly subjects are substantially slower in responding to a required modification of their movement than in responding to the required movement initiation. Patients with Parkinson's disease have impaired capabilities in processing simultaneously the motor responses to two visual stimuli presented in rapid succession. PMID- 9728946 TI - Dopaminergic stimulation in unilateral neglect. AB - OBJECTIVE: To explore the hypothesis that dopaminergic circuits play a part in the premotor components of the unilateral neglect syndrome, the effects of acute dopaminergic stimulation in patients with neglect were studied. METHODS: Two tasks were evaluated before and after subcutaneous administration of apomorphine and placebo: a circle crossing test and a test of target exploration (a modified version of the bell test), performed both in perceptual (counting) and in perceptual-motor (pointing) conditions. SUBJECTS: Four patients with left neglect. RESULTS: After dopaminergic stimulation, a significant improvement was found compared with placebo administration and baseline evaluation, in the performance of the two tests. Three of the patients had a more marked improvement in the perceptual-motor condition (pointing) of the task than the perceptual condition (counting). CONCLUSIONS: The findings suggest that dopaminergic neuronal networks may mediate, in different ways, both perceptive and premotor components of the unilateral neglect syndrome. PMID- 9728945 TI - Highly abnormal thermotests in familial dysautonomia suggest increased cardiac autonomic risk. AB - OBJECTIVE: Patients with familial dysautonomia have an increased risk of sudden death. In some patients with familial dysautonomia, sympathetic cardiac dysfunction is indicated by prolongation of corrected QT (QTc) interval, especially during stress tests. As many patients do not tolerate physical stress, additional indices are needed to predict autonomic risk. In familial dysautonomia there is a reduction of both sympathetic neurons and peripheral small nerve fibres which mediate temperature perception. Consequently, quantitative thermal perception test results might correlate with QTc values. If this assumption is correct, quantitative thermotesting could contribute to predicting increased autonomic risk. METHODS: To test this hypothesis, QTc intervals were determined in 12 male and eight female patients with familial dysautonomia, aged 10 to 41 years (mean 21.7 (SD 10.1) years), in supine and erect positions and postexercise and correlated with warm and cold perception thresholds assessed at six body sites using a Thermotest. RESULTS: Due to orthostatic presyncope, six patients were unable to undergo erect and postexercise QTc interval assessment. The QTc interval was prolonged (>440 ms) in two patients when supine and in two additional patients when erect and postexercise. Supine QTc intervals correlated significantly with thermal threshold values at the six body sites and with the number of sites with abnormal thermal perception (Spearman's rank correlation p<0.05). Abnormal Thermotest results were more frequent in the four patients with QTc prolongation and the six patients with intolerance to stress tests. CONCLUSION: The results suggest that impaired thermal perception correlates with cardiac sympathetic dysfunction in patients with familial dysautonomia. Thus thermotesting may provide an alternative, albeit indirect, means of assessing sympathetic dysfunction in autonomic disorders. PMID- 9728948 TI - Psychopathological and emotional deficits in myotonic dystrophy. AB - OBJECTIVE: To evaluate psychopathological disturbances in patients with myotonic dystrophy (MD) and compare patients with MD to both patients with facioscapulohumeral dystrophy (FSHD) and healthy control subjects. METHODS: A semistructured interview was used to determine DSM III-R criteria for major depressive episodes, dysthymic episodes, and generalised anxiety. The Montgomery and Asberg and the Hamilton depressive scales, the Covi and Tyrer anxiety scales, the Abrams and Taylor scale for emotional blunting, and the depressive mood scale were all used in the study. Subjects were also asked to complete questionnaires for physical and social anhedonia. RESULTS: Fifteen patients with MD, 11 patients with FSHD, and 14 healthy subjects were studied. Patients with MD were not more depressed or anxious than healthy controls. Patients with FSHD were the most depressed and most anxious. However, patients with MD had significantly lower scores for expressiveness and significantly higher scores for anhedonia than the other two groups. CONCLUSION: Patients with MD did not present significant depressive or anxious symptomatology but rather an emotional deficit. This emotional deficit may be an adaptive reaction to the threatening implications of the disease, or the effect of the CNS lesions which occur with MD, or both. PMID- 9728947 TI - Diagonal spatial neglect. AB - OBJECTIVE: To determine whether stroke patients with diagonal neglect on cancellation may show diagonal neglect on line bisection, and hence to indicate whether diagonal neglect may be related solely to the type of test used or whether instead it may reflect a fundamental spatial disorder. METHODS: Nine patients with subacute right hemispheric stroke who neglected targets primarily in the near left direction on line cancellation bisected diagonal lines of two opposing orientations: near left to far right and far left to near right. The errors were assessed to determine whether line orientation significantly affected bisection error. RESULTS: Eight patients had significant bisection errors. One of these showed no effect of line orientation on error, consistent with lateral neglect. The remaining seven patients had a line orientation effect, indicating a net diagonal spatial bias. For the group, cancellation errors were significantly correlated with the line orientation effect on bisection errors. CONCLUSIONS: A significant diagonal bias on two tests of spatial attention may appear in stroke patients, although the directions of the biases may differ within individual patients. None the less, diagonal neglect may be a fundamental spatial attentional disturbance of right hemispheric stroke. Greater severity of stroke deficit as indicated by cancellation error score may be associated with a greater degree of diagonal neglect on line bisection. PMID- 9728949 TI - Single fibre electromyography in multifocal motor neuropathy with persistent conduction blocks. AB - OBJECTIVE: To study the process of denervation-reinnervation in multifocal motor neuropathy with persistent conduction blocks in clinically affected and unaffected muscles. METHOD: Volitional single fibre electromyography (SFEMG) was performed in the extensor digitorum communis (EDC) of seven patients. The jitter, the fibre density, and the mean interpotential interval were determined. The results before and after treatment with intravenous immunoglobulin (IVIg) between the unaffected EDC and affected EDC examined during the same SFEMG session were also compared. In addition the values of jitter, fibre density, and mean interpotential interval were analysed for correlation with the strength score on the MRC scale, the duration of the neuropathy, the number of IVIg treatment periods, and the radial nerve conduction block values. RESULTS: Mean jitter, percentage of jitters >60 micros, and impulse blocking percentage, were higher than normal in both the affected EDCs and to a lesser degree in unaffected EDCs. Jitter decreased significantly after IVIg and correlated only with the MRC score. Fibre density and mean interpotential interval were higher than normal equally in the affected EDC and unaffected EDCs, but no correlation was found with strength, duration of the neuropathy, number of treatment periods, and conduction block values. CONCLUSION: The major finding is the presence of SFEMG abnormalities in clinically unaffected EDCs. This shows a process of denervation-reinnervation even in the absence of clinical symptoms, probably more frequent than commonly supposed in this neuropathy. The rapid clinical improvement after IVIg infusions could be due to remyelination after demyelination and to an interference of IVIg with the blocking effect of antibodies on the Na+ channels at the motor nerve endings. PMID- 9728950 TI - Use of corticosteroids in multiple sclerosis by consultant neurologists in the United Kingdom. AB - OBJECTIVES: To survey the use of corticosteroids in multiple sclerosis as recommended by United Kingdom consultant neurologists. METHODS: A postal questionnaire covering the use of corticosteroids for acute multiple sclerosis relapse and chronic progressive multiple sclerosis with regard to frequency of use, type of corticosteroid, and dosage regime was sent to all members of the Association of British Neurologists with a United Kingdom address. RESULTS: Two hundred and twelve United Kingdom consultant neurologists replied to the survey (74% response rate). Eighty six per cent indicated that they prescribed corticosteroids in more than one quarter of acute multiple sclerosis relapses seen. Intravenous methylprednisolone was recommended at some time by 99% of consultant neurologists, the most popular regime being 1g daily for 3 days (74%; 154/ 208). Over one half (53%; 109/206) never recommended a subsequent tapering course of oral corticosteroids; of those that did, 25% (24/96) recommended a tapering course lasting more than 1 month. Eighty eight per cent (1811206) of prescribers of intravenous methylprednisolone were able to offer the course as a day case on the ward; 7% (151206) at an outpatient clinic; and 5% (111206) at home. Almost three quarters of neurologists recommended oral corticosteroids for some acute relapses, although the most popular response was for occasional use only (48%; 1011212). Forty five per cent (961211) at least occasionally recommended steroids for patients with chronic multiple sclerosis not experiencing an acute relapse. CONCLUSIONS: Although the vast majority of consultant neurologists would prescribe intravenous methylprednisolone for acute multiple sclerosis relapse at some time, the use of corticosteroids for multiple sclerosis was otherwise variable. There seemed to be little consensus about the use of oral steroids in acute relapse, the prescribing of a tapering course of oral steroids after intravenous methylprednisolone, or the utility of steroids in chronic multiple sclerosis. Variability of prescribing recommendations probably reflects a lack of clear evidence in the face of a wide range of clinical situations, variable access, and timing of access to neurologists in the acute phase of relapse and pressure on neurologists to treat in an otherwise "hopeless" situation. Large multicentred trials are needed to consider these issues. PMID- 9728951 TI - Thoracocervicofacial purpura as the single manifestation of epileptic seizure. PMID- 9728952 TI - Sensory alien hand syndrome: case report and review of the literature. AB - An 81 year old right handed woman developed a left alien hand syndrome characterised by involuntary movements of choking and hitting the face, neck, and shoulder. The patient showed multiple disorders of primary sensation, sensory processing, hemispatial attention, and visual association, as well as a combination of sensory, optic, and cerebellar ataxia (triple ataxia) of the left arm in the absence of motor neglect or hemiparesis. Imaging studies disclosed subacute infarction in the right thalamus, hippocampus, inferior temporal lobes, splenium of corpus callosum, and occipital lobe due to right posterior cerebral artery occlusion. This rare syndrome should be considered as a "sensory" or "posterior" form of the alien hand syndrome, to be distinguished from the "motor" or "anterior" form described more commonly. PMID- 9728953 TI - Ataxia induced by small amounts of alcohol. AB - A patient is described who exhibited cerebellar ataxia after drinking small amounts of alcohol. Intake of 5 g alcohol induced a gaze evoked nystagmus, a scanning speech, a body sway after eye closure, and bilateral postural leg tremor. Kinematic and EMG analysis of fast wrist movements showed normal movements before and marked hypermetria after alcohol intake. Dysmetria was due to abnormal programming of antagonist muscle activity. PMID- 9728954 TI - Functional organisation of saccades and antisaccades in the frontal lobe in humans: a study with echo planar functional magnetic resonance imaging. AB - The cortical activation pattern of saccades and antisaccades (versus rest) in the frontal lobe was analysed using an echo planar imaging (EPI) technique in 10 healthy subjects. Statistical analysis of activity in the dorsolateral prefrontal cortex disclosed a significantly greater activation during antisaccades in this region than during saccades. On the other hand, activity in the frontal eye fields was not statistically different in both tasks. These results confirm the important role of the dorsolateral prefrontal cortex for the correct performance of antisaccades obtained by studies in humans with isolated lesions of the dorsolateral prefrontal cortex. PMID- 9728955 TI - The role of the alpha-synuclein gene mutation in patients with sporadic Parkinson's disease in the United Kingdom. AB - Parkinson's disease is a common neurodegenerative disorder of unknown aetiology. A pathogenic point mutation within the a-synuclein gene has recently been identified in one Italian-American kindred and three families of Greek origin with parkinsonism. DNA from 70 patients with Parkinson's disease was screened for this G209A mutation. No samples were positive for the mutation, suggesting that it is not relevant for most patients with sporadic idiopathic Parkinson's disease. PMID- 9728956 TI - Upbeat nystagmus: clinicoanatomical correlation. AB - A patient is reported on with isolated upbeating nystagmus with a linear slow phase in whom a solitary lesion, probably inflammatory, was detected radiologically in the dorsal paramedian caudal medulla, encompassing the most caudal of the perihypoglossal nuclei, the nucleus intercalatus of Staderini. The conjunction of a vestibular pattern of nystagmus with this focal lesion runs contrary to a previous suggestion that the nucleus intercalatus may act as a neural integrator for vertical conjugate eye movements. PMID- 9728958 TI - Atrial fibrillation and cognitive function: case-control study. AB - Atrial fibrillation is an important and independent risk factor for cerebrovascular disease and vascular dementia. There is increasing evidence that atrial fibrillation is associated with an increased risk of asymptomatic or silent cerebral infarction and as a result may confer an increased risk of progressive cognitive impairment on a person. In this study we sought to determine whether this hypothesis could be explored in a prospective case controlled design. Twenty seven patients with non-valvular atrial fibrillation (NVAF) and no history of stroke, transient ischaemic attack, dementia, and thyrotoxicosis were compared with 54 age and sex matched controls in sinus rhythm. All cases underwent clinical examination, ECG, and psychological assessment using a battery of nine neuropsychological tests. Between group analysis and a comparison of mean test scores of paired controls with cases were undertaken. The presence of atrial fibrillation was consistently associated with poorer performances on all the subtests of the neuropsychological battery. There was no association between duration of atrial fibrillation and performance. These results provide evidence to justify further examination of the hypothesis in a larger prospective study to determine whether antithrombotic therapy may protect against cognitive decline in patients at maximal risk of silent cerebral ischaemia and associated cognitive decline. PMID- 9728957 TI - Separate visual pathways for perception of actions and objects: evidence from a case of apperceptive agnosia. AB - Recognition of different kinds of visual stimuli was studied in a patient who acquired apperceptive visual agnosia after a bilateral occipitotemporal lesion which partially spared the primary visual cortex. Impairment in recognising static objects perceived visually sharply contrasts with the relatively well preserved ability to recognise objects from gestures illustrating their use, and to recognise actions shown in line drawings. It is suggested that the occipitoparieto-frontal pathway is involved in the recognition of actions, and in the recognition of objects when sensorimotor experience is evoked. PMID- 9728959 TI - Association learning in the acute confusional state. AB - The usefulness of cognitive rehabilitative treatment in the acute stages after brain injury seems questionable because patients in severe acute confusional state early after coma clinically seem unable to learn and store new information. Therefore, the capability of patients in acute confusional state to learn and retain associative information was assessed. On two occasions pairs of simple nouns were presented to six patients in severe acute confusional state. Stimuli were presented repeatedly either in written form only or with additional pictorial representations. Immediate and 20 minutes delayed recall was measured. Patients in acute confusional state were able to learn progressively more word pairs across several presentations. They retained some information over an interval of 20 minutes. In addition, they learned and remembered pictorially supported associations better than pure verbal associations. Patients in severe acute confusional state may retain some explicit information and may profit from an imagery mnemonic aid. These results were not expected on the basis of clinical findings alone and they have potential implications for the care of patients in acute confusional state. PMID- 9728960 TI - Neurotrophin-3 is increased in skin in human diabetic neuropathy. AB - Neurotrophin-3 (NT-3), a member of the neurotrophin family, has been shown to be necessary for the development of muscle spindle and Merkel cell afferent nerve fibres in animal models. The presence of NT-3 in the suprabasal epidermis, where many unmyelinated sensory fibres terminate, has been shown for the first time. As these fibres are affected in early diabetic neuropathy and a clinical trial of recombinant human NT-3 in diabetic neuropathy is in progress, the concentrations of endogenous NT-3 in skin of 24 patients at different stages of diabetic polyneuropathy have been investigated. NT-3 concentrations, measured with a specific immunoassay, were significantly higher in affected skin biopsies from patients with diabetic neuropathy than matched control skin (diabetic skin 6.32 (1.18) pg/mg v control skin 1.28 (0.05) (mean (SEM)); p<0.004, Mann-Whitney U test), particularly in the later stages. The optical density of NT-3 immunostaining was also significantly greater in the epidermis in diabetic patients (diabetic epidermis 0.30 (0.06) v controls 0.24 (0.01); p<0.02). No correlation was found between individual quantitative sensory tests and the increase of NT-3 concentration. The increase of NT-3 seems to reflect the degree of skin denervation in diabetic neuropathy, and may represent a compensatory mechanism. The concentrations of NT-3 in other peripheral targets deserve study in diabetic neuropathy. PMID- 9728961 TI - Fluid attenuation inversion recovery (FLAIR) images of dentatorubropallidoluysian atrophy: case report. AB - The white matter lesions in a patient with late adult onset dentatorubropallidoluysian atrophy (DRPLA) were studied in detail by MRI using the fluid attenuation inversion recovery (FLAIR) technique. The patient was a 60 year old woman with a family history of DRPLA, in whom the number of CAG repeats in the DRPLA gene on chromosome 12 was expanded to 59 (normal allele 10). In addition to atrophy of the cerebral cortex, cerebellum, and pontomesencephalic tegmentum, the cerebral white matter and a part of the white matter tracts within the brainstem showed prominent high signal intensities on FLAIR images. These MR findings suggest that, in addition to the degeneration of the dentatorubral and pallidoluysian systems, the pathological process extends to the white matter in DRPLA. This could be important for differentiating DRPLA from other clinically similar diseases such as Machado-Joseph disease or Huntington's disease. PMID- 9728962 TI - Cerebellar swelling and massive brain stem distortion: spontaneous resolution documented by MRI. PMID- 9728964 TI - Hereditary motor and sensory neuropathy type 1A associated with sensorineural deafness. PMID- 9728963 TI - Carbohydrate antigen 19-9 in cerebrospinal fluid and within malignant cells in a case of leptomeningeal carcinomatosis. PMID- 9728965 TI - Chronic inflammatory demyelinating polyneuropathy accompanied by carcinoma. PMID- 9728966 TI - Frequency, causes, and consequences of burns in patients with epilepsy. PMID- 9728967 TI - A quartet of Down's syndrome, Alzheimer's disease, cerebral amyloid angiopathy, and cerebral haemorrhage: interacting genetic risk factors. PMID- 9728968 TI - Chhabra hydrocephalus shunt: lessons for gravitational valves. PMID- 9728969 TI - A case of chronic paroxysmal hemicrania responding to subcutaneous sumatriptan. PMID- 9728970 TI - IgM anti-GM2 antibody in a patient with Guillain-Barre syndrome subsequent to cytomegalovirus hepatitis cross reacts with N-acetylgalactosaminyl GD1a. PMID- 9728971 TI - Electrophysiological recordings in bilateral hemifacial spasm. PMID- 9728972 TI - Depression and its relation to lesion location after stroke. PMID- 9728973 TI - Freedom of information, when it suits. PMID- 9728974 TI - No clear answers on safety of pigs as tissue donor source. PMID- 9728975 TI - Unravelling mysteries of neuroblastoma. PMID- 9728976 TI - FGFR2 mutations and acne. PMID- 9728977 TI - Can HCV infection be cleared? PMID- 9728978 TI - Uric acid as an independent risk factor in the treatment of hypertension. PMID- 9728979 TI - Unexpected endotracheal extubations. PMID- 9728980 TI - Methods for decreasing antibiotic use in otitis media. PMID- 9728982 TI - Randomised comparison of implantation of heparin-coated stents with balloon angioplasty in selected patients with coronary artery disease (Benestent II) AB - BACKGROUND: The multicentre, randomised Benestent-II study investigated a strategy of implantation of a heparin-coated Palmar-Schatz stent plus antiplatelet drugs compared with the use of balloon angioplasty in selected patients with stable or stabilised unstable angina, with one or more de-novo lesions, less than 18 mm long, in vessels of diameter 3 mm or more. METHODS: 827 patients were randomly assigned stent implantation (414 patients) or standard balloon angioplasty (413 patients). The primary clinical endpoint was event-free survival at 6 months, including death, myocardial infarction, and the need for revascularisation. The secondary endpoints were the restenosis rate at 6 months and the cost-effectiveness at 12 months. There was also one-to-one subrandomisation to either clinical and angiographic follow-up or clinical follow up alone. Analyses were by intention to treat. FINDINGS: Four patients (one stent group, three angioplasty group) were excluded from analysis since no lesion was found. At 6 months, a primary clinical endpoint had occurred in 53 (12.8%) of 413 patients in the stent group and 79 (19.3%) of 410 in the angioplasty group (p=0.013). This significant difference in clinical outcome was maintained at 12 months. In the subgroup assigned angiographic follow-up, the mean minimum lumen diameter was greater in the stent group than in the balloon-angioplasty group, (1.89 [SD 0.65] vs 1.66 [0.57] mm, p=0.0002), which corresponds to restenosis rates (diameter stenosis > or =50%) of 16% and 31% (p=0.0008). In the group assigned clinical follow-up alone, event-free survival rate at 12 months was higher in the stent group than the balloon-angioplasty group (0.89 vs 0.79, p=0.004) at a cost of an additional 2085 Dutch guilders (US$1020) per patient. INTERPRETATION: Over 12-month follow-up, a strategy of elective stenting with heparin-coated stents is more effective but also more costly than balloon angioplasty. PMID- 9728983 TI - Neuroblastoma in Europe: differences in the pattern of disease in the UK. SENSE. Study group for the Evaluation of Neuroblastoma Screening in Europe. AB - BACKGROUND: Neuroblastoma is a major contributor to childhood cancer mortality, but its prognosis varies with age and stage of disease, and some tumours regress spontaneously. Urinary screening programmes or clinical examination may detect the disease before symptoms appear, but the benefit of early diagnosis is uncertain. We examined the incidence, pattern, and presentation of neuroblastoma in four European countries. METHOD: Population-based incidence rates were derived for France, Austria, Germany, and the UK. Age, sex, and stage distribution were analysed by Mantel-Haenszel techniques and Poisson regression. The proportion of incidental diagnoses (cases without symptoms found at routine health checks or during investigation of other disorders) and mortality rates were also compared. FINDINGS: Between 1987 and 1991, 1672 cases of neuroblastoma were diagnosed in children under 15 years old (France, 624; Austria, 69; Germany, 493; UK, 486). Age-standardised annual incidence was significantly lower in the UK (10.1/million) than in France (12.5) and Germany (11.4). In the UK a deficit of low-stage disease in infants was accompanied by an excess of stage IV in older children. The UK had significantly fewer incidental diagnoses (8%) than Austria (27%) and Germany (34%). UK mortality rates were significantly higher than German or French rates. INTERPRETATION: In the UK, neuroblastoma diagnosis is delayed, possibly because of a less rigorous system of health checks for children. Although some overdiagnosis occurs in mainland Europe, our data suggest that in the UK some low-stage cases, undetected in infancy, may later present as advanced disease. This finding has implications for screening programmes and organisation of routine surveillance of infant health in the UK. PMID- 9728984 TI - Effect of long-chain polyunsaturated fatty acids in infant formula on problem solving at 10 months of age. AB - BACKGROUND: Long-chain polyunsaturated fatty acids (LCPUFA) are important for normal visual and brain development. Although present in human milk, LCPUFA have until recently been absent from artificial formulas, and infants may have limited ability to synthesise LCPUFA. To determine the clinical significance of this relative deficiency of LCPUFA, we undertook a randomised trial of the relation between LCPUFA supplementation and infant cognitive behaviour. METHODS: 44 term infants had been randomised to a formula supplemented with LCPUFA (21) or not supplemented with LCPUFA (23), which they had taken from birth to age 4 months. Infant cognitive behaviour was assessed at 10 months of age by a means-end problem-solving test--the intentional execution of a sequence of steps to achieve a goal. The problem required three intermediate steps to achieve the final goal, uncovering and retrieving a hidden toy. FINDINGS: Infants who received LCPUFA supplemented formula had significantly more intentional solutions than infants who received the no-LCPUFA formula (median 2.0 vs 0, p=0.021). Intention scores (median 14.0 vs 11.5 [maximum 18]) were also increased in this group (p=0.035). INTERPRETATION: These findings suggest that term infants may benefit from LCPUFA supplementation, and that the effects persist beyond the period of supplementation. Since higher problem-solving scores in infancy are related to higher childhood IQ scores, supplementation with LCPUFA may be important for the development of childhood intelligence. PMID- 9728985 TI - Expression of pig endogenous retrovirus by primary porcine endothelial cells and infection of human cells. AB - BACKGROUND: The risk of interspecies transmission of retroviruses during xenotransplantation is suggested by reports of pig endogenous retrovirus (PERV) released from porcine cell lines productively infecting human cell lines in vitro and of infectious PERV being released from pig peripheral blood mononuclear cells after mitogenic stimulation. Endothelial cells are the main interface between a xenograft and the recipient's leucocytes and tissues. METHODS: We have analysed pig primary aortic endothelial cells (PAEC) together with other transplantation relevant porcine cells and tissues for expression of PERV mRNA. Release of virus particles by PAEC was monitored by reverse transcriptase (RT) activity in the medium of cultured PAEC. Infectivity for human cells was tested by co-cultivation of irradiated PAEC with the human embryonal kidney cell line HEK293 and looking for virus release from the human cells. FINDINGS: PAECs, hepatocytes, lung, and skin from a variety of pig strains and breeds expressed PERV mRNA. PAEC released infectious particles. Co-cultivation of PAEC and HEK293 led to productive infection of the human cells and expression of PERV types A and B. INTERPRETATION: Release of infectious virus from PAEC occurred without mitogenic stimulation, suggesting a serious risk of retrovirus transfer after xenotransplantation. PMID- 9728986 TI - No evidence of infection with porcine endogenous retrovirus in recipients of porcine islet-cell xenografts. AB - BACKGROUND: The study of whether porcine xenografts can lead to porcine endogenous retrovirus (PERV) infection of recipients is critical for evaluating the safety of pig-to-man xenotransplantation. PERV is carried in the pig germline, and all recipients of porcine tissues or organs will be exposed to the virus. METHODS: We studied 10 diabetic patients who had received porcine fetal islets between 1990 and 1993, looking for evidence of PERV infection by using PCR serology, PCR, and reverse transcriptase assays. Prolonged xenograft survival (up to a year) was confirmed in five patients by porcine C-peptide excretion and detection of pig mitochondrial DNA (mtDNA) in serum. FINDINGS: Despite the evidence for extended exposure to pig cells and despite concomitant immunosuppressive therapy, we were unable to detect markers of PERV infection in any patient. Screening for two PERV sequences in peripheral blood lymphocytes collected 4-7 years after the xenotransplantation was negative. Markers of PERV expression, including viral RNA and reverse transcriptase, were undetectable in sera from both early (day 3 to day 180) and late (4-7 years) time points. Western blot analysis for antibodies was consistently negative. INTERPRETATION: These results suggested the absence of PERV infection in these patients. Also this study establishes a minimum standard for post-transplant surveillance of patients given porcine xenografts. PMID- 9728987 TI - No evidence of pig DNA or retroviral infection in patients with short-term extracorporeal connection to pig kidneys. AB - BACKGROUND: The xenotransplantation of organs and tissues, in particular those from pigs, is viewed as a means to alleviate the shortage of human donor organs and cells available for transplantation and also as a therapy for other diseases. The potential microbiological hazards of xenotransplantation have recently attracted much attention. One concern is over pig endogenous retroviruses (PERV). Until the possible consequences of infection by PERV are better understood it is unlikely that a significant number of porcine xenotransplants will proceed. However, a small number of patients have already been treated with or exposed to living porcine cells or tissue, and investigation of these patients may provide valuable information. METHODS: We took serial blood samples from two renal dialysis patients whose circulation had been linked extracorporeally to pig kidneys and tested them for pig DNA and PERV DNA by nested PCR. The patients' plasma was also tested for neutralising antibodies to two anthropotropic PERV strains. FINDINGS: Having established that the nested PCRs could detect single molecules of target sequence, we analysed DNA isolated from patients' peripheral blood mononuclear cells. We found no evidence of pig or PERV DNA in either patient, even in samples taken as early as 6 h after the perfusion. Furthermore, we found no evidence of seroconversion for PERV-specific antibodies. INTERPRETATION: The absence of porcine cells in the circulation of both patients, even in the samples taken soon after the perfusion experiment, suggests that any porcine cells dislodged from the kidney became rapidly sequestered from the circulation. Since cell-to-cell contact increases the efficiency of infection of PERV this removal of porcine cells may increase the risk of transmission of PERV to the xenograft recipient. We did not, however, detect indications of infection by PERV by PCR or neutralisation assay. The genetic and serological methods described here will be useful for detection of possible PERV infection in other patients. PMID- 9728988 TI - Hands up for angina. PMID- 9728989 TI - Prion immunoreactivity in appendix before clinical onset of variant Creutzfeldt Jakob disease. PMID- 9728990 TI - Epidermal mosaicism producing localised acne: somatic mutation in FGFR2. PMID- 9728991 TI - Shuttle-walk test to assess chronic heart failure. PMID- 9728992 TI - Treatment of Tourette's syndrome with mecamylamine. PMID- 9728993 TI - Outbreak of meningococcal disease linked to a sports club. PMID- 9728994 TI - Fetal exposure to maternal cortisol. PMID- 9728995 TI - Symptomatic restenosis after carotid percutaneous transluminal angioplasty. PMID- 9728996 TI - Cultured epidermal-sheet graft for epidermodysplasia verruciformis. PMID- 9728997 TI - Ultraviolet B and blood pressure. PMID- 9728998 TI - Patient-detected diurnal changes in spleen volume. PMID- 9728999 TI - Regimen lowers rate of coronary events in diabetes. PMID- 9729000 TI - Robert Moellering: protecting the public health. PMID- 9729001 TI - Bringing surgery to the rural areas of Ecuador. PMID- 9729002 TI - US food-safety system needs major overhaul. PMID- 9729003 TI - Compensation for mental illness resulting from Holocaust trauma. PMID- 9729004 TI - Primary pulmonary hypertension. AB - Primary pulmonary hypertension (PPH) is a progressive disease characterised by raised pulmonary vascular resistance, which results in diminished right-heart function due to increased right ventricular afterload. PPH occurs most commonly in young and middle-aged women; mean survival from onset of symptoms is 2-3 years. The aetiology of PPH is unknown, although familial disease accounts for roughly 10% of cases, which suggests a genetic predisposition. Current theories on pathogenesis focus on abnormalities in interaction between endothelial and smooth-muscle cells. Endothelia-cell injury may result in an imbalance in endothelium-derived mediators, favouring vasoconstriction. Defects in ion-channel activity in smooth-muscle cells in the pulmonary artery may contribute to vasoconstriction and vascular proliferation. Diagnostic testing primarily excludes secondary causes. Catheterisation is necessary to assess haemodynamics and to evaluate vasoreactivity during acute drug challenge. Decrease in pulmonary vascular resistance in response to acute vasodilator challenge occurs in about 30% of patients, and predicts a good response to chronic therapy with oral calcium-channel blockers. For patients unresponsive during acute testing, continuous intravenous epoprostenol (prostacyclin, PGI2) improves haemodynamics and exercise tolerance, and prolongs survival in severe PPH (NYHA functional class III-IV). Thoracic transplantation is reserved for patients who fail medical therapy. We review the progress made in diagnosis and treatment of PPH over the past 20 years. PMID- 9729005 TI - Intellectual property, drug licensing, freedom of information, and public health. PMID- 9729006 TI - Whitehall gets quality supporter. Interview by Kelly Morris. PMID- 9729007 TI - The UDHR and the physician's role. PMID- 9729008 TI - Health, memory, and human rights in Guatemala. PMID- 9729009 TI - The Taliban's war on women. PMID- 9729010 TI - Support for Mbarara Hospital. PMID- 9729011 TI - Support for Mbarara Hospital. PMID- 9729012 TI - Support for Mbarara Hospital. PMID- 9729013 TI - Support for Mbarara Hospital. PMID- 9729014 TI - Support for Mbarara Hospital. PMID- 9729015 TI - Support for Mbarara Hospital. PMID- 9729016 TI - Breastfeeding versus formula feeding in HIV infection. PMID- 9729017 TI - Adjuvant therapy of melanoma. PMID- 9729018 TI - Sorting the hype from the facts in melanoma. PMID- 9729019 TI - Detection of retroviral antibodies in primary biliary cirrhosis. PMID- 9729020 TI - Antipyretic therapy in acute stroke. PMID- 9729021 TI - Antipyretic therapy in acute stroke. PMID- 9729022 TI - Antipyretic therapy in acute stroke. PMID- 9729023 TI - Peanut allergy. PMID- 9729024 TI - Peanut allergy. PMID- 9729025 TI - The 12th world AIDS conference. PMID- 9729026 TI - Prevention of tuberculosis in HIV-1. PMID- 9729028 TI - Mortality among street youth in the UK. PMID- 9729027 TI - Drug treatment for Crohn's disease. PMID- 9729029 TI - Measuring quality in the NHS. PMID- 9729030 TI - Multiple hearts in animals other than Barosaurus. PMID- 9729031 TI - Getting hooked on marmite. PMID- 9729032 TI - Adoption of hospital policies for prevention of perinatal group B streptococcal disease--United States, 1997. AB - Group B streptococcal (GBS) infections are the leading bacterial cause of disease and deaths among newborns in the United States. In 1993, the annual cost of caring for newborns with sepsis caused by group B Streptococcus was an estimated $294 million. A survey of hospital GBS disease prevention practices in 1994 indicated that those hospitals with a prenatal screening policy had fewer neonatal GBS disease cases. In 1996, to promote a coordinated approach to prevention, CDC issued consensus guidelines about GBS disease prevention that were endorsed by the American College of Obstetricians and Gynecologists and the American Academy of Pediatrics. These consensus guidelines recommend using either a screening-based strategy or a risk-based strategy for identifying women who should receive intrapartum antimicrobial prophylaxis. To evaluate adoption of the consensus guidelines, in 1997, hospitals in eight surveillance areas were surveyed, and the results were compared with findings of a similar survey conducted in 1994. The proportion of hospitals with prevention policies in each site was compared with the site's rate of early-onset disease to assess the impact of the prevention policies. This report presents the survey findings, which indicate that more hospitals have adopted GBS disease prevention policies since issuance of the consensus guidelines. PMID- 9729033 TI - Rural health-care providers' attitudes, practices, and training experience regarding intimate partner violence--West Virginia, March 1997. AB - Primary health-care providers are an important resource to women who have experienced intimate partner violence (IPV). IPV patients in rural areas often face obstacles to preventive services such as physical isolation from health care and a lack of adequate community services. In March 1997, a pilot project was conducted to survey the attitudes and practices of rural health-care providers (RHCPs) toward women at risk for IPV in primary-care clinics in West Virginia and to determine the training experience of RHCPs in IPV intervention and prevention during the preceding 2 years. This report summarizes the results of the survey, which indicate that most RHCPs recognized barriers to identification and referral of abused women in their practices but few screened female patients for IPV or have had recent continuing education on IPV. PMID- 9729034 TI - Update: leptospirosis and unexplained acute febrile illness among athletes participating in triathlons--Illinois and Wisconsin, 1998. AB - Since July 14, 1998, the Illinois Department of Health, the Wisconsin Department of Health, the U.S. Department of Agriculture (USDA), and CDC, in collaboration with other state and local health departments, have been investigating an outbreak of acute febrile illness among athletes from 44 states and seven countries who participated in triathlons in Springfield, Illinois, on June 21, 1998, and in Madison, Wisconsin, on July 5, 1998. Initial testing at CDC of specimens from four athletes identified leptospirosis as the illness in all four. This report updates the ongoing investigation of this outbreak through August 13, which indicates that Leptospira was the etiologic agent for illness in athletes and in persons with occupational or recreational exposure to Lake Springfield, where the event was held in Illinois. PMID- 9729035 TI - Regulation of telomere length and telomerase in T and B cells: a mechanism for maintaining replicative potential. PMID- 9729036 TI - Developmentally programmed rearrangement of T cell receptor Vgamma genes is controlled by sequences immediately upstream of the Vgamma genes. AB - Distinct subsets of gammadelta T cells expressing different Vgamma and Vdelta chains arise in ordered waves during thymic development. In the murine Jgamma1 Cgamma1 cluster, the Vgamma3 gene segment is utilized earliest in fetal thymic development, in progenitors of dendritic epidermal T cells (DECs). The Vgamma2 gene segment predominates in the late fetal stages and beyond, in cells destined for the secondary lymphoid organs. Using transgenic TCRgamma recombination substrates, we demonstrate that this restricted Vgamma gene usage is determined by developmentally targeted gene rearrangement. We show that sequences immediately upstream of the Vgamma2 and Vgamma3 genes direct the rearrangement pattern in adult thymocytes. Thus, the choice of Vgamma gene for recombination is coordinated with distinct differentiation programs in gammadelta subsets. PMID- 9729037 TI - Individual variations in the murine T cell response to a specific peptide reflect variability in naive repertoires. AB - Previous studies have analyzed the diversity of T cell responses upon immunization. Little is known, however, about the individual variability of naive repertoires and its influence on immune responses. In the present study, T cells specific for a Kd-restricted epitope derived from HLA-A2 were purified from individual immunized mice using tetramers of MHC-peptide. Their TCRbeta chains were sequenced revealing strong biases but large variations in BJ usage and clonal composition. Most importantly, sequence analysis from nonimmunized mice demonstrated the preexistence of a small set of splenic precursors, distinct in each mouse and comprising less than 200 cells. Therefore, differences in precursor pools appear to be the major source of individual variability in antigen-selected repertoires. PMID- 9729038 TI - A molecular map of T cell development. AB - Using a sensitive molecular marker for positive selection, the appearance of a particular functional TCR alpha chain sequence in cells from mice bearing a transgenic beta chain, we address several aspects of intrathymic T cell development. First, by examining specific TCR prior to and after maturation, we demonstrate how a restricted TCR repertoire is positively selected from a highly diverse immature TCR repertoire. Second, since this molecular marker is enriched in cells progressing toward the CD4 lineage and depleted in cells progressing toward the CD8 lineage, a map of the developmental pathway of alphabeta thymocytes can be inferred. Third, the first cells that show clear signs of positive intrathymic selection are identified. PMID- 9729039 TI - Requirement for the thymus in alphabeta T lymphocyte lineage commitment. AB - We recently identified a fetal thymic developmental stage (NK1.1+/CD117(lo)) that characterizes committed T/NK progenitors. We now report the existence of phenotypically and functionally identical T/NK progenitors in mouse fetal blood and spleen but not in fetal liver. These precursors are indistinguishable from previously characterized fetal blood "prothymocytes" (CD90+/CD117(lo)), with the exception that they express NK1.1, lack markers associated with T lineage commitment, maintain a germline TCRbeta locus, and can give rise to both T and NK cells. Moreover, NK1.1+/CD90+/CD117(lo) fetal blood precursors are present in athymic nude mice. These results suggest that the T/NK lineage commitment pathway is thymus-independent. In contrast, full commitment to the alphabeta T lineage does not precede thymus colonization. PMID- 9729040 TI - The crystal structure of H-2Dd MHC class I complexed with the HIV-1-derived peptide P18-I10 at 2.4 A resolution: implications for T cell and NK cell recognition. AB - The structure of H-2Dd complexed with the HIV-derived peptide P18-I10 (RGPGRAFVTI) has been determined by X-ray crystallography at 2.4 A resolution. This MHC class I molecule has an unusual binding motif with four anchor residues in the peptide (G2, P3, R/K/H5, and I/L/F9 or 10). The cleft architecture of H 2Dd includes a deep narrow passage accomodating the N-terminal part of the peptide, explaining the obligatory G2P3 anchor motif. Toward the C-terminal half of the peptide, p5R to p8V form a type I' reverse turn; residues p6A to p9T, and in particular p7F, are readily exposed. The structure is discussed in relation to functional data available for T cell and natural killer cell recognition of the H 2Dd molecule. PMID- 9729041 TI - Mice with a fluorescent marker for interleukin 2 gene activation. AB - Production of interleukin (IL)-2 by T lymphocytes is one of the earliest events during immune response. A mutant mouse strain was generated by replacing the IL-2 gene with a cDNA encoding green fluorescent protein (GFP). In this model, GFP fluorescence is readily detectable upon T cell activation and is mostly coexpressed with IL-2 at the single cell level. Thus, individual activated T cells can express the IL-2 gene biallelically. Upon stimulation through the T cell antigen receptor, CD4+ cells separate into distinct GFP+ and GFP- populations, both of which are capable of differentiating into either Th1 or Th2 effectors. These mice allow noninvasive detection of IL-2 production by single cells and analysis of the subsequent differentiative fate of these cells as an immune response develops. PMID- 9729042 TI - Regulation of IL-4 expression by activation of individual alleles. AB - To study the in vivo role of IL-4-expressing cells, we developed a strategy to tag these cells, by generating mice in which one IL-4 allele was replaced with a cDNA encoding the human CD2 (huCD2) cell-surface molecule. Expression of the huCD2 reporter was, like IL-4, restricted to the appropriately polarized T helper 2 cells. However, most of the cells expressed only the IL-4 or the targeted allele. Analysis of the frequency of monoallelic versus biallelic expression suggests that the activation of each individual allele is regulated by a stochastic process whose probability can be augmented by increasing the strength of signal delivered through the TCR. Allele-specific activation may be a general feature of cytokine regulation that contributes to the functional diversity within T helper cell subpopulations. PMID- 9729043 TI - Helper T cell differentiation is controlled by the cell cycle. AB - Helper T (Th) cell differentiation is highly regulated by cytokines but initiated by mitogens. By examining gene expression in discrete generations of dividing cells, we have delineated the relationship between proliferation and differentiation. Initial expression of IL-2 is cell cycle-independent, whereas effector cytokine expression is cell cycle-dependent. IFNgamma expression increases in frequency with successive cell cycles, while IL-4 expression requires three cell divisions. Cell cycle progression and cytokine signaling act in concert to relieve epigenetic repression and can be supplanted by agents that hyperacetylate histones and demethylate DNA. Terminally differentiated cells exhibit stable epigenetic modification and cell cycle-independent gene expression. These data reveal a novel mechanism governing Th cell fate that initially integrates proliferative and differentiative signals and subsequently maintains stability of the differentiated state. PMID- 9729044 TI - LAT palmitoylation: its essential role in membrane microdomain targeting and tyrosine phosphorylation during T cell activation. AB - The linker molecule LAT is a critical substrate of the tyrosine kinases activated upon TCR engagement. Phosphorylated LAT binds Grb2, PLC-gamma1, and other signaling molecules. We demonstrate that human LAT is palmitoylated and that palmitoylated LAT predominantly localizes into glycolipid-enriched microdomains (GEMs). Although the LAT transmembrane domain is sufficient for membrane localization, palmitoylation at C26 and C29 is essential for efficient partitioning into GEMs. LAT palmitoylation is necessary for its tyrosine phosphorylation. After T cell activation, most tyrosine-phosphorylated LAT molecules and a fraction of PLC-gamma1 and other signaling molecules are present in GEMs. LAT is central to T cell activation and is a novel linker molecule shown to require targeting to membrane microdomains for signaling. PMID- 9729045 TI - CD30 is a CD40-inducible molecule that negatively regulates CD40-mediated immunoglobulin class switching in non-antigen-selected human B cells. AB - We used our monoclonal model of germinal center maturation, CL-01 B cells, to investigate the role of CD30 in human B cell differentiation. CL-01 cells are IgM+ IgD+ CD30+ and switch to IgG, IgA, and IgE when exposed to CD40L and IL-4. Switching is hampered by CD30 coengagement, possibly through interference with the CD40-mediated NF-kappaB-dependent transcriptional activation of downstream C(H) genes. The physiological relevance of this phenomenon is emphasized by similar CD30-mediated effects in naive B cells. Expression of CD30 by these cells is induced by CD40L but is inhibited by B cell receptor coengagement and/or exposure to IL-6 and IL-12. Our data suggest that CD30 critically regulates the CD40-mediated differentiation of non-antigen-selected human B cells. PMID- 9729046 TI - Probing immunoglobulin gene hypermutation with microsatellites suggests a nonreplicative short patch DNA synthesis process. AB - As the rate of Ig gene hypermutation approximates the level of nucleotide discrimination of DNA polymerases (10(-3) to 10(-4)), a local inhibition of proofreading and mismatch repair during semiconservative replication could generate the mutations introduced by the process. To address this question, we have constructed transgenic mice that carry a hypermutation substrate containing a "polymerase slippage trap": an Ig gene with a mono or dinucleotide tract inserted in its V region. The low amount of slippage events as compared to the number of mutations, the absence of transient misalignment mutations at the border of the repeats, and the dissociation between the amount of frameshifts and mutations when the transgene is put on mismatch repair-deficient genetic backgrounds, suggest that Ig gene hypermutation occurs by an error-prone short patch DNA synthesis taking place outside global DNA replication. PMID- 9729047 TI - Targeted disruption of the mouse Caspase 8 gene ablates cell death induction by the TNF receptors, Fas/Apo1, and DR3 and is lethal prenatally. AB - Homozygous targeted disruption of the mouse Caspase 8 (Casp8) gene was found to be lethal in utero. The Caspase 8 null embryos exhibited impaired heart muscle development and congested accumulation of erythrocytes. Recovery of hematopoietic colony-forming cells from the embryos was very low. In fibroblast strains derived from these embryos, the TNF receptors, Fas/Apo1, and DR3 were able to activate the Jun N-terminal kinase and to trigger IkappaB alpha phosphorylation and degradation. They failed, however, to induce cell death, while doing so effectively in wild-type fibroblasts. These findings indicate that Caspase 8 plays a necessary and nonredundant role in death induction by several receptors of the TNF/NGF family and serves a vital role in embryonal development. PMID- 9729048 TI - Intracellular neutralization of HIV transcytosis across tight epithelial barriers by anti-HIV envelope protein dIgA or IgM. AB - Human immunodeficiency virus, generated during contact between HIV-infected cells and the apical surface of an epithelial cell, can cross a tight epithelial barrier by transcytosis. We show that transcytosis of primary HIV isolates is blocked by dimeric IgA or IgM against HIV envelope proteins. Neutralization occurs intracellularly within the apical recycling endosome, and immune complexes are specifically recycled to the mucosal surface. One epitope involved in neutralization is a conserved sequence of the gp41 HIV envelope protein subunit. Finally, transcytosis also occurs across functional human mucosal tissue in a process inhibited by a serosal internalization of IgM against the HIV envelope protein. These results suggest that induction of mucosal immunity to HIV envelope proteins may impair the transcytotic route of HIV mucosal transmission. PMID- 9729049 TI - Simulators of malignant melanoma of the skin. AB - Early recognition of malignant melanoma is crucial in order to remove lesions at a stage when complete cure can still be achieved. Unfortunately, however, some melanomas defy clinical and/or histopathological recognition, and some benign lesions (melanocytic or non-melanocytic) can present with clinical and/or histopathological aspects simulating malignant melanoma. Awareness of these simulators is important to avoid pointless (and potentially dangerous) aggressive treatments. In this review, we have illustrated the clinicopathological features of the most important benign simulators of malignant melanoma of the skin. PMID- 9729050 TI - An in vitro strategy to evaluate the phototoxicity of solar UV at the molecular and cellular level: application to photoprotection assessment. AB - Skin cancers are among the most common human cancers and have an increasing incidence. The ultraviolet radiation components of sunlight play a major role in skin tumor induction and development. Cellular DNA has been identified as a target for most of the biological effects of UV, and the induction of photodamage is considered as the initiating step of photocarcinogenesis. Thus, effective photoprotection of DNA against harmful overex-posure to solar UV is a critical issue. The efficiency of a sunscreen is usually tested with respect to its ability to prevent skin erythema, but conceivably, more data are required at the molecular and cellular level in order to ascertain protection against photocarcinogenic risk. In the present study, we define a strategy based on the use of various in vitro models and solar-simulated light to evaluate photodamage and photoprotection: -Supercoiled circular plasmid DNA for detection of structural alterations. -The yeast Saccharomyces cerevisiae to evaluate cytotoxicity and genotoxicity. -The single-cell gel electrophoresis or comet assay to determine DNA damage and DNA repair in human keratinocytes. -p53 expression as a hallmark for genotoxic stress. -Induction of pigmentation in human melanocytes. In conditions where light source, spectrum and control of radiation delivery were precisely defined, we have demonstrated that the wide spectrum UVA sunscreen Mexoryl SX protects from the cytotoxicity and genotoxicity of solar UV. PMID- 9729051 TI - Primary cutaneous nocardiosis. AB - A 37-year-old patient with systemic lupus erythematosus, who had been treated with oral corticosteroids for 10 years, developed primary cutaneous nocardiosis. Brown-violaceous, suppurative nodules and plaques arose on her right leg. Cultures of multiple biopsies on blood agar medium grew Nocardia. The patient received 500 mg of imipenem three times a day which resulted in complete regression of the lesions within two months. No relapse was observed 6 months later. PMID- 9729052 TI - Detection of HHV-8 DNA in a German patient with classical Kaposi's sarcoma may allow an estimation of the incubation period. AB - DNA of HHV-8 (Kaposi's sarcoma-associated Herpes virus [KSHV]) was detected in a biopsy of a Kaposi's sarcoma in an elderly male patient from Saxony (East Germany). The diagnosis of classical Kaposi's sarcoma was first made in 1986. During World War II, the patient had been on active service on the Greek Islands of Crete and Rhodes only, he did not travel outside East Germany after the war. It is assumed that the patient was infected during his stay on the islands of Crete or Rhodes, where classical Kaposi's sarcoma is endemic. If so, the incubation period of classical Kaposi's sarcoma could be as long as 40 years. PMID- 9729053 TI - Basal cell destruction of herpes gestationis and bullous pemphigoid. PMID- 9729054 TI - Mometasone furoate decreases adhesion molecule expression in psoriasis. AB - The topical corticosteroids are widely used in the treatment of moderate psoriasis, because of their usefulness for reducing inflammation and controlling itching. The therapeutic effect of corticosteroids in different cutaneous inflammatory diseases may be partially explained by their varying ability to block in vitro the synthesis of different cytokines, which play a pivotal role in epidermal hyperproliferation and leukocyte recruitment into the skin. The purpose of the present investigation was to further elucidate the mode of action of mometasone furoate, a medium-high potency, topical corticosteroid, on adhesion molecules, cytokines and cytokine receptor expression in psoriatic skin. Using an immunohistochemical assessment, we examined lesional skin biopsies from ten psoriatic patients before treatment and after 1 and 3 weeks of therapy. The overexpression of alpha 2, alpha 3, alpha 6, and beta 1 integrins detected in the spinous layer of untreated psoriatic skin was significantly decreased after therapy in 8 out of 10 cases, characterized by only partial clinical remission. In the remaining patients, a disappearance of the above integrin reactivity paralleling the disappearance of psoriatic lesions was induced by the treatment. With the exception of GM-CSF, no or only marginal effects of mometasone furoate on the cytokine and cytokine receptor system were observed. A significant reduction of the positive immunostaining with anti-ICAM-1 and ICAM-2 monoclonal antibodies on dermal vascular endothelial cells was also seen. Thus, our findings indicate that the therapeutic effects of mometasone furoate in psoriasis are mediated principally by decreasing adhesion molecule expression and to a lesser degree by inhibiting cytokine synthesis. PMID- 9729055 TI - Bullous prurigo pigmentosa and diabetes. AB - Prurigo pigmentosa (PP) is a type of inflammatory dermatosis characterized by pruritic, reddish, papular lesions that normally resolve while leaving gross reticular pigmentation. In severe cases however, they may form edematous infiltrative plaques, but no formation of vesicles or bullae is generally found. We herein present the case of a 32-year-old Japanese male patient with diabetes mellitus, who developed a severe vesicular formation. Minocycline was found to be very effective. In addition, the eruption subsided when the urine glucose and ketone levels were controlled by glibenclamide. The most characteristic feature in this case was the fact that numerous vesicles and bullae were seen both in the beginning and throughout the clinical course. It therefore seems that a sudden exacerbation of diabetes mellitus was associated with a severe formation of vesicles and bullae. The findings of this case may suggest a correlation between diabetes mellitus and PP. PMID- 9729056 TI - Contact pemphigus induced by dihydrodiphenyltrichlorethane. AB - We describe a case of contact pemphigus, a new subgroup of induced pemphigus. The disease is provoked by a chlorinated hydrocarbon pesticide: dihydrodiphenyltrichlorethane. We suspect that systemic absorption after the topical contact is responsible for the alteration of skin structure and activation of immunological mechanisms leading to blister formation and acantholysis. PMID- 9729057 TI - Pathways of development of human dendritic cells. PMID- 9729058 TI - Langerhans cells are susceptible to measles virus infection and actively suppress T cell proliferation. AB - We have previously reported that the measles virus (MV) could productively infect human dendritic cells (DC), derived in vitro from CD34+ cord blood progenitors in the presence of GM-CSF and TNF-alpha. In this study, we provide evidence that freshly isolated Langerhans cells (LC), which are immature dendritic cells located in skin and mucosal epithelia, are susceptible to MV infection in vitro as assessed by viral antigen expression by both LC syncytia and LC remaining as single cells. Moreover, MV-infected LC completely block naive allogeneic CD4+ T cell proliferation in response to uninfected LC. This active inhibitory effect is not due to virus transmission from infected LC, is independent of the maturation stage of LC at the time of infection and is antigen non-specific and MHC unrestricted. Thus, both immature and mature LC are susceptible to measles virus infection, which turns these efficient antigen-presenting cells into active tolerogenic cells. LC infection may explain the long lasting immune suppression observed during natural measles infection. PMID- 9729059 TI - Generalized granuloma annulare and hepatitis B vaccination. AB - As hepatitis B vaccination is becoming generalized in Europe, cutaneous adverse events are being more frequently reported in the literature. We report the first case of generalized granuloma annulare following hepatitis B immunization. A 51 year-old woman presented a generalized granuloma annulare one month after the one year booster injection of the hepatitis B vaccine. The lesions resolved with sulfone therapy. We observed an identical recurrence three weeks after the five year booster. PMID- 9729060 TI - Horse bite injury. AB - Bite wounds are relatively frequent, the order of frequency being, dogs, cats and humans. The clinical importance of other types of bites depends on the severity of the injury or any subsequent infection. We report on the case of a woman bitten on her thigh by a horse, producing severe haematoma, fat necrosis and muscle rupture, without an external wound. We emphasize the importance of the ultrasound examination in the evaluation of the extent of the crush injury. PMID- 9729061 TI - Photosensitive drug eruption induced by flutamide. AB - We describe a 68-year-old man who showed a photosensitive drug eruption induced by flutamide. The minimal erythema dose with ultraviolet A light was reduced to 2 J/cm2 under administration of flutamide, which recovered to over 16 J/cm2 after cessation of the medication, without changing the reactivity to ultraviolet B. The absorption spectrum of flutamide was not altered after ultraviolet A irradiation, suggesting that flutamide is photostable with regard to ultraviolet A. In addition, bovine serum albumin was not covalently photocoupled with flutamide under ultraviolet A. Thus, flutamide seems to have low potency to act as a photohapten, and it is likely that a non-photohaptenic mechanism operates in this photosensitivity or its active metabolite may work as a photosensitizer. PMID- 9729062 TI - Dyskeratosis congenita associated with Hodgkin's disease. AB - Dyskeratosis congenita is a rare, hereditary, multisystem disorder characterized by mucocutaneous changes, pancytopenia and increased incidence of malignancy. Different types of neoplasia have been reported in association with dyskeratosis congenita. We present a second case associated with Hodgkin's disease. Delayed appearance of dermatological signs and association with chronic hepatitis B are other unusual features of this case. PMID- 9729063 TI - Esophageal foreign bodies. PMID- 9729064 TI - The promise of reward. PMID- 9729065 TI - Impact of temporal variation and the balance between excitation and inhibition on the output of the perfect integrate-and-fire model. AB - We consider how the output of the perfect integrate-and-fire (I&F) model of a single neuron is affected by the properties of the input, first of all by the distribution of afferent excitatory and inhibitory postsynaptic potential (EPSP, IPSP) inter-arrival times, discriminating particularly between short- and long tailed forms, and by the degree of balance between excitation and inhibition (as measured by the ratio, r, between the numbers of inhibitory and excitatory inputs). We find that the coefficient of variation (CV; standard deviation divided by mean) of efferent interspike interval (ISI) is an increasing function of the length of the tail of the distribution of EPSP inter-arrival times and the ratio r. There is a range of values of r in which the CV of output ISIs is between 0.5 and 1. Too tight a balance between EPSPs and IPSPs will cause the model to produce a CV outside the interval considered to correspond to the physiological range. Going to the extreme, an exact balance between EPSPs and IPSPs as considered in [24] ensures a long-tailed ISI output distribution for which the moments such as mean and variance cannot be defined. In this case it is meaningless to consider quantities like output jitter, CV, etc. of the efferent ISIs. The longer the tail of the input inter-arrival time distribution, the less is the requirement for balance between EPSPs and IPSPs in order to evoke output spike trains with a CV between 0.5 and 1. For a given short-tailed input distribution, the range of values of r in which the CV of efferent ISIs is between 0.5 and 1 is almost completely inside the range in which output jitter (standard deviation of efferent ISI) is greater than input jitter. Only when the CV is smaller than 0.5 or the input distribution is a long-tailed one is output less than input jitter [21]. The I&F model tends to enlarge low input jitter and reduce high input jitter. We also provide a novel theoretical framework, based upon extreme value theory in statistics, for estimating output jitter, CV and mean firing time. PMID- 9729066 TI - Three-dimensional structure of the ion-channel forming peptide trichorzianin TA VII bound to sodium dodecyl sulfate micelles. AB - Trichorzianin TA VII, Ac0 U1 A2 A3 U4 J5 Q6 U7 U8 U9 S10 L11 U12 P13 V14 U15 I16 Q17 Q18 Fol19, is a nonadecapeptide member of the peptaibol antibiotics biosynthesized by Trichoderma soil fungi, which is characterized by a high proportion of the alpha, alpha-dialkylated amino acids, alpha-aminoisobutyric acid (Aib, U) and isovaline (Iva, J), an acetylated N-terminus and a C-terminal phenylalaninol (Pheol, Fol). The main interest in such peptides stems from their ability to interact with phospholipid bilayers and form voltage-dependent transmembrane channels in planar lipid bilayers. In order to provide insights into the lipid-peptide interaction promoting the voltage gating, the conformational study of TA VII in the presence of perdeuterated sodium dodecyl sulfate (SDS-d25) micelles has been carried out. 1H sequential assignment have been performed with the use of two-dimensional homo- and -heteronuclear nmr techniques including double quantum filtered correlated spectroscopy, homonuclear Hartmann-Hahn, nuclear Overhauser effect spectroscopy, 1H-13C heteronuclear single quantum correlation, and heteronuclear multiple bond correlation. Conformational parameters, such as 3JNHC alpha H coupling constants, temperature coefficients of amide protons (delta gamma/delta TNH) and quantitative nuclear Overhauser enhancement data, lead to detailed structural information. Ninety eight three-dimensional structures consistent with the nmr data were generated from 231 interproton distances six phi dihedral angle restraints, using restrained molecular dynamics and energy minimization calculations. The average rms deviation between the 98 refined structures and the energy-minimized average structure is 0.59 A for the backbone atoms. The structure of trichorzianin TA VII associated with SDS micelles, as determined by these methods, is characterized by two right-handed helical segments involving residues 1-8 and 11-19, linked by a beta-turn that leads to an angle about 90 degrees-100 degrees between the two helix axes; residues 18 and 19 at the end of the C-terminal helix exhibit multiple conformations. PMID- 9729067 TI - IInd Miniaturisation in Liquid Chromatography versus Capillary Electrophoresis Conference. Gent, Belgium, 26-28 May 1997. Extended abstracts. PMID- 9729068 TI - Breast implants deemed safe--again. PMID- 9729069 TI - Blood supplies to be treated to reduce CJD risk. PMID- 9729070 TI - Fluid resuscitation with colloid or crystalloid solutions. Newer synthetic colloids should not be abandoned. PMID- 9729071 TI - Fluid resuscitation with colloid or crystalloid solutions. One conclusion could be that hypertonic saline is better than colloids in trauma. PMID- 9729072 TI - Fluid resuscitation with colloid or crystalloid solutions. Eight studies should have been excluded. PMID- 9729073 TI - Fluid resuscitation with colloid or crystalloid solutions. Use of dextran-70 for fluid resuscitation has been dying out. PMID- 9729075 TI - Fluid resuscitation with colloid or crystalloid solutions. Virtually identical article had appeared in Cochrane Library. PMID- 9729074 TI - Fluid resuscitation with colloid or crystalloid solutions. Conditions and patient groups were too heterogeneous to allow meaningful comparisons. PMID- 9729076 TI - Communication among health professionals. Message pagers may improve communication. PMID- 9729077 TI - Should industry sponsor research? Tobacco company sponsorship discredits medical but not all research. PMID- 9729078 TI - Should industry sponsor research? Condemning the drinks industry rules out potentially useful research. PMID- 9729079 TI - Should industry sponsor research? If the drinks industry does not clean up its act, pariah status is inevitable. PMID- 9729080 TI - Should industry sponsor research? Collaborative research with infant formula companies should not always be censored. PMID- 9729081 TI - Should industry sponsor research? How much research in infant feeding comes from unethical marketing? PMID- 9729082 TI - Passive smoking and heart disease. Evidence on passive smoking and heart disease needs re-evaluation. PMID- 9729083 TI - Passive smoking and heart disease. BMJ should encourage open debate of available evidence. PMID- 9729084 TI - Passive smoking and heart disease. There must be better uses for money spent on vilifying passive smoking. PMID- 9729085 TI - Passive smoking and lung cancer. Risk extrapolation overestimates risk. PMID- 9729087 TI - The hot air on passive smoking. BAT has not tried to discredit data on passive smoking. PMID- 9729086 TI - Passive smoking and lung cancer. Accumulated evidence on lung cancer and environmental tobacco smoke. PMID- 9729088 TI - The hot air on passive smoking. Opinions depend on what sort of evidence is thought most convincing. PMID- 9729089 TI - Marketing of breast milk substitutes. Italy has initiatives regarding compliance with international code. PMID- 9729090 TI - Marketing of breast milk substitutes. Manufacturers have sponsored healthcare journals. PMID- 9729091 TI - Action on human albumin not swift enough. PMID- 9729092 TI - Dutch to examine testing boxers for brain damage. PMID- 9729093 TI - Vitamin B-6 reprieved in UK. PMID- 9729094 TI - Commentary: Meta-analysis is a blunt and potentially misleading instrument for analysing models of service delivery. PMID- 9729095 TI - Neonatal screening for cystic fibrosis. Early diagnosis allows option of prenatal diagnosis in subsequent pregnancies. PMID- 9729096 TI - Neonatal screening for cystic fibrosis. Early diagnosis is important to parents even if it makes little difference to outcome. PMID- 9729097 TI - Health beliefs among British Bangladeshis. Health promotion for Bangladeshi women in general practice must be appropriate. PMID- 9729098 TI - Partnership with patients. Telephone helpline services can meet patients' demand for information. PMID- 9729099 TI - Management of deliberate self poisoning. Psychiatric services have limited role in deliberate self poisoning. PMID- 9729100 TI - Towards an understanding of integrative brain functions. Analysis at multiple levels. Proceedings of the Nobel Symposium 103. Stockholm, Sweden, 4-6 June 1997. PMID- 9729101 TI - Pharmaceutical laboratory compliance--manufacturer's view. PMID- 9729102 TI - Report recognizes greater role for vitamin supplements, fortified foods. PMID- 9729103 TI - Mitochondrial presequences. PMID- 9729105 TI - Homeopathic treatment for sequelae of meningococcal septicemia. PMID- 9729106 TI - Periodontal Diseases and Human Health: New Directions in Periodontal Medicine. Papers Presented at the March 1997 Sunstar-Chapel Hill Symposium. Chapel Hill, North Carolina, March 24-25, 1997. PMID- 9729104 TI - Proceedings of a National Symposium on Environmental Contaminants and the Implications for Child Health. Ottawa, Ontario, Canada. May 25-27, 1997. PMID- 9729107 TI - Change in bacterial community during biodegradation of aniline. AB - The response of river water microbial communities to chemical compounds was monitored under laboratory conditions using aniline as a model. Bacteria were collected from unpolluted and polluted sites. Bacterial abundance (plate and total direct counting) and its relation to aniline biodegradation was examined. Colony hybridization with 16S rRNA oligonucleotide probes was used to study the changes in microbial community structure during biodegradation of aniline. The changes in bacterial abundance and community structure were related to biodegradation of aniline. Burkholderia-Pseudomonas (rRNA group III), an authentic Alcaligenes group became dominant despite the initial differences in the microbial communities, suggesting that these genera are the main aniline degraders in the aquatic environment. PMID- 9729108 TI - Smoking: an uncommon research topic. PMID- 9729109 TI - Comment on the workshop on Airway Inflammation and Remodelling in Asthma: implication for asthma therapy. PMID- 9729110 TI - Transgenic fish and its application in basic and applied research. AB - Since 1985, transgenic fish have been successfully produced by microinjecting or electroporating desired foreign DNA into unfertilized or newly fertilized eggs using many different fish species. More recently, transgenic fish have also been produced by infecting newly fertilized eggs with pantropic, defective retroviral vectors carrying desired foreign DNA. These transgenic fish can serve as excellent experimental models for basic scientific investigations as well as in biotechnological applications. In this paper, we will review the current status of the transgenic fish research and its potential application in basic and applied research. PMID- 9729111 TI - A case of tuberculosis meningitis complicated by intracerebral hemorrhage. PMID- 9729112 TI - Rhabdomyolysis following acute ischemic stroke. PMID- 9729113 TI - Complex partial status epilepticus provoked by hyponatremia. PMID- 9729114 TI - Cerebral fat embolism: debated acute posttraumatic encephalography. PMID- 9729115 TI - Herpes zoster ophthalmicus and delayed contralateral hemiparesis: a case of ipsilateral midbrain involvement. PMID- 9729116 TI - Spinal epidural abscess: treatment options. PMID- 9729117 TI - The ordeal of chronic pain. PMID- 9729118 TI - Identification and genomic organization of a gene coding for a new member of the cell adhesion molecule family mapping to Xq25. AB - The gene coding for a new member of the Immunoglobulin (Ig)-like domain containing molecule superfamily has been identified and mapped to the human Xq25 chromosomal band. It contains 12 Ig-like domains in two clusters of 5 and 7 motifs, respectively, separated by a linker segment, followed by a transmembrane and a cytoplasmic region. The gene is conserved in mammals and is expressed in muscle, heart, brain, testis, and pancreas with transcripts of different length, suggesting that it is subjected to alternative processing. The transcript is assembled from 19 exons which are distributed along approx. 20kb; each Ig-like domain is contained in distinct exons which constitute the unit of repeated genomic duplications. Elucidation of the IGDC1 genomic structure will allow the investigation of the basis of its alternative transcription and of its possible involvement in diseases mapped to the Xq25 interval. PMID- 9729119 TI - Characterization of an element positively regulating the transcription of MSSP gene-2 which encodes C-MYC binding proteins. AB - MSSP gene-2 encodes at least three alternative splicing products, MSSP-1, MSSP-2, and Scr2, which have been implied to function as factors regulating DNA replication, transcription, apoptosis induction, and cell-cycle movement, via the interaction with C-MYC. We have previously shown that the Mup1 sequence from -474 to -440 in the region required for transactivation of the MSSP gene-2 was protected from DNaseI digestion in vitro. Here, we report that the Mup1 sequence carries a positive transcriptional activity, and that the activity was lost by point mutations that inhibited the formation of specific nucleoprotein complexes on the sequence. In Southwestern analyses, two proteins of 38 and 62kDa directly interacted with the Mup1 sequence. PMID- 9729120 TI - Molecular analysis of the P97 cilium adhesin operon of Mycoplasma hyopneumoniae. AB - Mycoplasma hyopneumoniae causes an economically significant respiratory disease of swine called Enzootic Pneumonia. The disease process is initiated by adherence of M. hyopneumoniae to the cilia of swine respiratory epithelium through an interaction involving P97, a surface-associated protein, and cilia-specific receptors. Binding specificity is associated with a repeat region located near the C-terminus of the P97 protein. Further analysis of the DNA sequences surrounding the P97 structural gene revealed an operon composed of two ORFs, P97 and one coding for a 102.3-kDa protein designated P102. Hybridization analysis and subcloning experiments showed that the P97 adhesin-encoding gene was present as a single copy in the M. hyopneumoniae chromosome. P102 sequences, however, were found on four distinct chromosomal fragments, suggesting that multiple copies of P102 were present in the chromosome. One of these clones was identified by screening the genomic library with swine convalescent sera showing that P102 is expressed in vivo during M. hyopneumoniae infections. All copies of P102 were mapped to a single chromosomal region comprising approximately 13% of the genome (140kb), although the exact distance between the copies is not known. The function of P102 is also not known, but the translated sequence shows a prominent transmembrane domain, suggesting that it may be a surface protein. PMID- 9729121 TI - Molecular cloning of a GPI-anchored aminopeptidase N from Bombyx mori midgut: a putative receptor for Bacillus thuringiensis CryIA toxin. AB - An aminopeptidase N (APN) with a molecular weight of 110kDa was released from the midgut membrane of Bombyx mori by phosphatidylinositol-specific phospholipase C (PI-PLC), and purified to a homogeneous state. This 110-kDa APN was different from the 100-kDa APN that we previously reported, in chromatographic behaviors, substrate specificity, and N-terminal and internal amino acid sequences. However, the N-terminal sequence of 110-kDa APN, DPAFRLPTTTRPRHYQVTLT, was highly homologous with those of Manduca sexta and Heliothis virescens APNs, which were identified as a receptor for an insecticidal toxin of Bacillus thuringiensis. From a B. mori midgut cDNA library, we cloned the 110-kDa APN cDNA that possessed a 2958-bp open reading frame encoding a 111573-Da polypeptide of 986 residues. The sequence of the eicosa-peptide Asp42Thr61 deduced from the cDNA was completely matched with the N-terminal sequence of the mature 110-kDa APN. One potential N-glycosylation site, HEXXHXW zinc-binding motif and characteristic proline-rich repeats were observed in the ORF. Moreover, the primary sequence contained two hydrophobic peptides on N- and C-termini. The N-terminal peptide sequence showed characteristics of leader peptide for secretion and the C terminal peptide contained a possible glycosylphosphatidylinositol (GPI) anchoring site. Taken together, the deduced amino acid sequence suggests that the 110-kDa APN is a GPI-anchored protein and a specific receptor protein for B. thuringiensis CryIA delta-endotoxin. PMID- 9729122 TI - Structure of the mouse gene for the serine protease inhibitor neuroserpin (PI12). AB - Neuroserpin (PI12), initially identified as an axonally secreted protein in cultured chicken dorsal root ganglion neurons, belongs to the serpin family of the serine protease inhibitors and is mainly expressed by neurons of both the developing and the adult nervous system. Here we report on the cloning and structural characterization of the neuroserpin gene of the mouse. The murine neuroserpin gene spans over more than 55kb and consists of nine exons. The positions and phases of the exonintron borders are completely conserved between neuroserpin and its nearest homologues, protease nexin-1 and plasminogen activator inhibitor-1. A single transcription initiation site, which is colocalized with a potential initiation (Inr) sequence, has been determined by primer extension and RNase protection. Sequence analysis revealed a TATA-less promoter with a CAAT box and several sites for the general transcription factor Sp1 and the neuron-specific transcription factor AP-2. PMID- 9729123 TI - Cloning and expression of the gene coding for FtsH protease from Mycobacterium tuberculosis H37Rv. AB - This study was aimed at the molecular cloning and expression of the gene coding for FtsH protease of Mycobacterium tuberculosis H37Rv (virulent). PCR on the genomic DNA of M. tuberculosis H37Ra (non-virulent) using the oligodeoxynucleotide primers, which were designed based on the codon usage pattern of M. tuberculosis and against the nucleotide (nt) sequence corresponding to two conserved domains of the FtsH protein of Escherichia coli, yielded a 363 bp product. The amino-acid sequence, deduced from the nt sequence of the PCR product, revealed the presence of two ATP-binding motifs and the AAA Signature motif (Second Region of Homology) that are characteristic features found conserved in the FtsH molecules from eubacteria, archaebacteria, and eukaryotes. Southern hybridisation of the NheI digest of the cosmid SCY6F7 containing part of the genomic DNA of M. tuberculosis H37Rv using the PCR fragment as the probe identified the full-length ftsH gene in the 7.2-kb fragment. The gene was subcloned into pBS (SK+) vector, and the FtsH product that was expressed in E. coli transformed with the vector was identified as an 85-kDa protein localised in the membrane. PMID- 9729124 TI - Genomic structure of the human congenital chloride diarrhea (CLD) gene. AB - Congenital chloride diarrhea (CLD) is caused by mutations in a gene which encodes an intestinal anion transporter. We report here the complete genomic organization of the human CLD gene which spans approximately 39kb, and comprises 21 exons. All exon/intron boundaries conform to the GT/AG rule. An analysis of the putative promoter region sequence shows a putative TATA box and predicts multiple transcription factor binding sites. The genomic structure was determined using DNA from several sources including multiple large-insert libaries and genomic DNA from Finnish CLD patients and controls. Exon-specific primers developed in this study will facilitate mutation screening studies of patients with the disease. Genomic sequencing of a BAC clone H_RG364P16 revealed the presence of another, highly homologous gene 3' of the CLD gene, with a similar genomic structure, recently identified as the Pendred syndrome gene (PDS). PMID- 9729126 TI - The 2nd Israel-Britain Conference on Forensic Psychiatry. London, United Kingdom, September/October 1996. Abstracts. PMID- 9729125 TI - [Morphological characteristics of esophageal cancer and its significance]. PMID- 9729127 TI - 5th Research Workshop on the Biology, Prevention, and Treatment of Head and Neck Cancer. Abstracts. PMID- 9729128 TI - Role of surfactant and hyperoxia. PMID- 9729129 TI - The Evolving Role of Retinoids in the Management of Cutaneous Conditions. New York, New York, USA. May 2-4, 1997. Conference proceedings. PMID- 9729130 TI - [Symposium on bronchogenic carcinoma. Clinical Center of the University of Sarajevo. Sarajevo, 7-8 November 1994]. PMID- 9729131 TI - A tribute to Duncan Neuhauser, PhD. PMID- 9729132 TI - Corticosteroids for the complications of Ross River virus infection. PMID- 9729133 TI - The 5th International Conference on Mechanisms of Antimutagenesis and Anticarcinogenesis. PMID- 9729134 TI - Antigenotoxicity studies in Drosophila melanogaster. AB - The fruit fly Drosophila melangaster with its well developed array of genotoxicity test systems has been used in a number of studies on antigenotoxicity of various compounds and mixtures. In recent years, the newly developed Somatic Mutation and Recombination Tests (SMART) have mainly been employed. These one-generation tests make use of the wing or eye imaginal disc cells in larvae and have proven to be very efficient and sensitive. They are based on the principle that the loss of heterozygosity of suitable recessive markers can lead to the formation of mutant clones of cells that are then expressed as spots on the wings or eyes of the adult flies. We have employed the wing spot test with the two markers multiple wing hairs (mwh,3-0.3) and flare (flr,3-38.8). Three-day-old larvae, trans-heterozygous for these markers, are treated chronically or acutely by oral administration with the test compound(s) or complex mixtures. For antigenotoxicity studies, chronic co-treatments can be used, as well as separate pre-treatments with an antigenotoxic agent followed by a chronic treatment with a genotoxin. After eclosion, the wings of the adult flies are scored for the presence of single and twin spots. These spots can be due to different genotoxic events: either mitotic recombination or mutation (deletion, point mutation, specific types of translocation, etc.). The analysis of two different genotypes (one with structurally normal chromosomes, one with a multiply inverted balancer chromosome) allows for a quantitative determination of the recombinagenic activity of genotoxins. Results of two separate studies presented: (1) instant coffee has antirecombinagenic but not antimutagenic activity in the wing spot test; and (2) ascorbic acid and catechin are able to protect against in vivo nitrosation products of methyl urea in combination with sodium nitrite. PMID- 9729135 TI - [31st Scientific meeting of the German-speaking Association of Mycology. Aachen, 18-21 September 1997]. PMID- 9729136 TI - Sildenafil in the treatment of erectile dysfunction. PMID- 9729137 TI - Sildenafil in the treatment of erectile dysfunction. PMID- 9729138 TI - Sildenafil in the treatment of erectile dysfunction. PMID- 9729139 TI - Sildenafil in the treatment of erectile dysfunction. PMID- 9729140 TI - Sildenafil in the treatment of erectile dysfunction. PMID- 9729141 TI - Sildenafil in the treatment of erectile dysfunction. PMID- 9729142 TI - Sildenafil in the treatment of erectile dysfunction. PMID- 9729143 TI - Lovastatin for X-linked adrenoleukodystrophy. PMID- 9729144 TI - Cancer and venous thromboembolism. PMID- 9729145 TI - Comparison of beclomethasone, salmeterol, and placebo in children with asthma. PMID- 9729146 TI - Hepatitis A vaccination. PMID- 9729147 TI - Treatment of pain in dying patients. PMID- 9729148 TI - Off at the races--a graphic patient history. PMID- 9729149 TI - [Primary Central Nervous System Lymphoma. Round table proceedings. Poitiers, France, 11-12 October 1996]. PMID- 9729151 TI - 50 Years of Pediatrics: 1948-1998. PMID- 9729150 TI - [Remembering Jozsef Polya (1802-1873)]. PMID- 9729152 TI - The birth and evolution of Pediatrics. PMID- 9729153 TI - The ever-changing content of Pediatrics over fifty years. PMID- 9729154 TI - The "fourth part" of Pediatrics: 50 years of display advertising. PMID- 9729155 TI - 50 years of Pediatrics: 1948-1998. The journal in 1947 and 1997: a dramatic change. PMID- 9729156 TI - Biographical sketches of the first editorial board and those who have edited Pediatrics. PMID- 9729157 TI - The growth and expansion of Pediatrics: 1948 to 1998. PMID- 9729158 TI - Reinventing a specialty: how Pediatrics survived its own success. PMID- 9729159 TI - The rational use of antimicrobials in acute otitis media: a bacteriologic investigation of otitis media in infancy, by E.A. Mortimer Jr, and R.L. Watterson Jr, Pediatrics, 1956;17:359-366; Otitis media in the practice of pediatrics: bacteriological and clinical [title truncated]. PMID- 9729160 TI - On the treatment of cerebral palsy: the outcome of 177 patients, 74 totally untreated, by Richmond Paine, MD, Pediatrics, 1962;29:605-616. PMID- 9729161 TI - Age limits of pediatrics, American Academy of Pediatrics, Council on Child Health, Pediatrics, 1972;49:463. PMID- 9729162 TI - Landmarks in child abuse and neglect: three flowers in the desert. The whiplash shaken infant syndrome, by J. Caffey, Pediatrics, 1974;54:396-403; Covert video recordings of life-threatening child abuse: lessons for child protection, by David P. Southall et al, Pediatrics, 1997;100:735-760; Preventing child abuse and neglect: a randomized trial of nurse home visitation, by David L. Olds et al, Pediatrics, 1986;78:65-78. PMID- 9729163 TI - Reye's syndrome and salicylate use, by Karen M. Starko, MD, et al, Pediatrics, 1980;66:859-864; and National patterns of aspirin use and Reye syndrome reporting, United States, 1980 to 1985, by Janet B. Arrowsmith et al, Pediatrics, 1987;79:858-863. PMID- 9729165 TI - Roberto Caldeyro-Barcia--an appreciation. PMID- 9729164 TI - CDF-1 mediated repression of cell cycle genes targets a specific subset of transactivators. AB - The cdc25C, cyclin A and cdc2 genes are regulated during the cell cycle through two contiguous repressor binding sites, the CDE and CHR, located in the region of transcription initiation and interacting with a factor termed CDF-1. The target of this repression seems to be transcriptional activation of these promoters by transcription factors bound upstream. The majority of these factors falls into the class of glutamine-rich activators, suggesting that CDF-1-mediated repression might be activation domain specific. In the present study we have used chimeric promoter constructs to demonstrate that the cdc25C UAS, but not the core promoter, is crucial for repression. In addition, we show that only specific transcription factors and activation domains are responsive to CDE-CHR-mediated cell cycle regulation. These observations clearly indicate that CDF-1 interferes with activation of transcription by a specific subset of transactivators. The repressible activation domains belong to the same class of glutamine-rich activators, pointing to specific interactions of CDF-1 with components of the transcription machinery. In agreement with this conclusion we find that a simple inversion of the CDF-CHR module completely abrogates cell cycle-regulated repression. PMID- 9729167 TI - The electronic Plant Gene Register. PMID- 9729166 TI - The control of flowering time and floral identity in Arabidopsis. PMID- 9729168 TI - [Central sterilization]. PMID- 9729170 TI - [Dissertations defended in 1998]. PMID- 9729169 TI - Unknown case #1. A 61-year-old women who had undergone a renal transplant in 1991 reported back and bilateral leg pain. PMID- 9729171 TI - [The 40th anniversary of Kamyshin Military Hospital]. PMID- 9729172 TI - Omeprazole, nitrenidipine, famotidine and stress-induced ulcers. A case of duplicate publication. PMID- 9729173 TI - Proceedings of the International Taurine Symposium '97: Cellular and Regulatory Mechanisms. Tucson, Arizona, USA. July 15-19, 1997. PMID- 9729174 TI - Hospitals and the family physician. PMID- 9729175 TI - Hospitalist concept: another dangerous trend. PMID- 9729176 TI - In memoriam: Marcus S. Goldstein (1906-1997). PMID- 9729177 TI - The 1st Bordeaux Basic and Clinical Research Meeting on Alcohol, Liver, and Nutrition: Utility of the Enteral (Tsukamoto-French) Alcohol Feeding Model. Bordeaux, France, September 22-25, 1997. PMID- 9729178 TI - Flock worker's lung: chronic interstitial lung disease in the nylon flocking industry. AB - BACKGROUND: Two young men working at a nylon flocking plant in Rhode Island developed interstitial lung disease of unknown cause. Similar clusters at the same company's Canadian plant were reported previously. OBJECTIVE: To define the extent, clinicopathologic features, and potential causes of the apparent disease outbreak. DESIGN: Case-finding survey and retrospective cohort study. SETTING: Academic occupational medicine program. PATIENTS: All workers employed at the Rhode Island plant on or after 15 June 1990. MEASUREMENTS: Symptomatic employees had chest radiography, pulmonary function tests, high-resolution computed tomography, and serologic testing. Those with unexplained radiographic or pulmonary function abnormalities underwent bronchoalveolar lavage, lung biopsy, or both. The case definition of "flock worker's lung" required histologic evidence of interstitial lung disease (or lavage evidence of lung inflammation) not explained by another condition. RESULTS: Eight cases of flock worker's lung were identified at the Rhode Island plant. Three cases were characterized by a high proportion of eosinophils (25% to 40%) in lavage fluid. Six of the seven patients who had biopsy had histologic findings of nonspecific interstitial pneumonia, and the seventh had bronchiolitis obliterans organizing pneumonia. All seven of these patients had peribronchovascular interstitial lymphoid nodules, usually with germinal centers, and most had lymphocytic bronchiolitis and interstitial fibrosis. All improved after leaving work. Review of the Canadian tissue specimens showed many similar histologic findings. Among the 165-member study cohort, a 48-fold or greater increase was seen in the sex-adjusted incidence rate of all interstitial lung disease. CONCLUSIONS: Work in the nylon flocking industry poses substantial risk for a previously unrecognized occupational interstitial lung disease. Nylon fiber is the suspected cause of this condition. PMID- 9729179 TI - Is colonoscopy indicated for small adenomas found by screening flexible sigmoidoscopy? AB - BACKGROUND: There is controversy over whether patients who have a small tubular adenoma on screening flexible sigmoidoscopy should undergo colonoscopic examination of the proximal colon. OBJECTIVE: To prospectively determine the prevalence of advanced polyps in the proximal colon among patients who have small adenomas on screening sigmoidoscopy. DESIGN: Prospective cohort study. SETTING: A health maintenance organization and a Veterans Affairs medical center. PATIENTS: Asymptomatic patients older than 50 years of age who had no risk factors for colon cancer and underwent sigmoidoscopy. INTERVENTION: At the time of sigmoidoscopy, all polyps were biopsied and characterized. All patients with distal adenomas were offered colonoscopy. MEASUREMENTS: The size and histology of polyps identified by sigmoidoscopy and colonoscopy were noted. Polyps were considered advanced if they were larger than 10 mm or were tubulovillous, villous, or malignant. The prevalence of advanced proximal polyps was determined, and patients were stratified by the size and number of distal polyps found by sigmoidoscopy. RESULTS: Among 4490 patients who underwent sigmoidoscopy, a neoplastic lesion was detected in 401 (8.9%) and colonoscopy was done in 301 (75%). Of 90 patients with a single tubular adenoma 1 to 5 mm in diameter in the distal colon, 0% (95% CI, 0.0% to 4.0%) had an advanced proximal polyp compared with 5.4% (CI, 2.4% to 10.4%) of those who had multiple distal polyps 1 to 5 mm or 6 to 10 mm in diameter and 7.9% (CI, 2.6% to 17.6%) of those who had advanced distal polyps (P = 0.013 for trend). The low-risk group with a single tubular adenoma 1 to 5 mm in diameter represented 44% of all patients with distal adenomas or cancers found at flexible sigmoidoscopy. CONCLUSIONS: Among patients undergoing screening sigmoidoscopy, those with single tubular adenomas of 5 mm or less had a low prevalence of advanced proximal polyps. These patients may not benefit from colonoscopy. PMID- 9729180 TI - Supraventricular arrhythmia in patients having noncardiac surgery: clinical correlates and effect on length of stay. AB - BACKGROUND: Few recent data are available on risk factors for perioperative supraventricular arrhythmia (SVA) after noncardiac surgery or on the effect of SVA on clinical outcomes. OBJECTIVE: To determine the incidence, clinical correlates, and effect on length of stay of perioperative SVA in patients having major noncardiac surgery. DESIGN: Prospective cohort study. SETTING: Urban tertiary care teaching hospital. PARTICIPANTS: 4181 patients 50 years of age or older who had major, nonemergency, noncardiac procedures and were in sinus rhythm at the preoperative evaluation. MEASUREMENTS: Preoperative clinical data, postoperative enzyme data, serial electrocardiograms, and clinical outcomes were collected prospectively. Outcomes were 1) SVA that persisted or led to treatment and 2) increase in length of stay attributable to SVA. RESULTS: Perioperative SVA occurred in 317 patients (7.6%); it occurred in 83 patients (2.0%) during surgery and in 256 (6.1%) after surgery. Independent preoperative correlates of SVA were male sex (odds ratio [OR], 1.3 [95% CI, 1.0 to 1.7]), age 70 years or older (OR, 1.3 [CI, 1.0 to 1.7]), significant valvular disease (OR, 2.1 [CI, 1.2 to 3.6]), history of SVA (OR, 3.4 [CI, 2.4 to 4.8]) or asthma (OR, 2.0 [CI, 1.3 to 3.1]), congestive heart failure (OR, 1.7 [CI, 1.1 to 2.7]), premature atrial complexes on preoperative electrocardiography (OR, 2.1 [CI, 1.3 to 3.4]), American Society of Anesthesiologists class III or IV (OR, 1.4 [CI, 1.1 to 1.9]), and type of procedure: abdominal aortic aneurysm (OR, 3.9 [CI, 2.4 to 6.3]) or abdominal (OR, 2.5 [CI, 1.7 to 3.6]), vascular (OR, 1.6 [CI, 1.1 to 2.4]), and intrathoracic (OR, 9.2 [CI, 6.7 to 13]) procedures. Among patients who had intrathoracic surgery, those receiving digoxin were at lower risk (OR, 0.2 [CI, 0.04 to 0.8]) for SVA than those not receiving digoxin. Patients with perioperative acute cardiac and noncardiac events had high relative risks for SVA. Supraventricular arrhythmia was associated with a 33% increase in length of stay after adjustment for other clinical data (P < 0.001). CONCLUSIONS: In this cohort, SVA was common after noncardiac surgery and was associated with prolonged length of stay. PMID- 9729181 TI - Clinical, hemodynamic, and cardiopulmonary exercise test determinants of survival in patients referred for evaluation of heart failure. AB - BACKGROUND: Accurate prognosis in chronic heart failure has become increasingly important in assessing the efficacy of treatment and in appropriately allocating scarce resources for transplantation. Previous studies of severe heart failure have been limited by short follow-up periods and few deaths. OBJECTIVE: To establish clinical, hemodynamic, and cardiopulmonary exercise test determinants of survival in patients with heart failure. DESIGN: Retrospective study. SETTING: Hospital-based outpatient heart failure clinic. PARTICIPANTS: 644 patients referred for evaluation of heart failure over 10 years. MEASUREMENTS: Age, cause of heart failure, body surface area, cardiac index, ejection fraction, pulmonary capillary wedge pressure, left ventricular dimensions, watts achieved during exercise, heart rate, maximum systolic blood pressure, and oxygen uptake (VO2) at the ventilatory threshold and at peak exercise were measured at baseline. Univariate and multivariate analyses were done for clinical, hemodynamic, and exercise test predictors of death. A Cox hazards model was developed for time of death. RESULTS: During a mean follow-up period of 4 years, 187 patients (29%) died and 101 underwent transplantation. Actuarial 1-year and 5-year survival rates were 90.5% and 73.4%, respectively. Resting systolic blood pressure, watts achieved, peak VO2, VO2 at the ventilatory threshold, and peak heart rate were greater among survivors than among nonsurvivors. Cause of heart failure (coronary artery disease or cardiomyopathy) was a strong determinant of death (relative risk for coronary artery disease, 1.73; P< 0.01). By multivariate analysis, only peak VO2 was a significant predictor of death. Stratification of peak VO2 above and below 12, 14, and 16 mL/kg per minute demonstrated significant differences in risk for death, but each cut-point predicted risk to a similar degree. CONCLUSIONS: Peak VO2 outperforms clinical variables, right-heart catheterization data, exercise time, and other exercise test variables in predicting outcome in severe chronic heart failure. Direct measurement of VO2 should be included when clinical or surgical decisions are being made in patients referred for evaluation of heart failure or those considered for transplantation. PMID- 9729182 TI - Prevalence of monoclonal gammopathies in patients with hepatitis C virus infection. AB - BACKGROUND: An association between monoclonal gammopathies and chronic liver diseases has been reported. OBJECTIVE: To determine the prevalence of monoclonal gammopathies in patients with chronic hepatitis C virus (HCV) infection and the possible association of monoclonal gammopathies with HCV genotypes. DESIGN: Prospective study. SETTING: Departments of internal medicine and hematology at two university hospitals in Italy. PATIENTS: 239 HCV-positive and 98 HCV-negative patients with chronic liver diseases were recruited consecutively. MEASUREMENTS: Clinical data were gathered, liver histologic examination was done, serum immunoglobulin and cryoglobulin levels were measured, and immunoelectrophoresis was done for monoclonal component detection. Patients with monoclonal gammopathy had serum HCV RNA measured and HCV genotype determined by polymerase chain reaction and had histologic examination of bone marrow. RESULTS: Monoclonal band was detected in 11% of HCV-positive patients and in 1% of HCV-negative patients (P = 0.004). The prevalence of HCV genotype 2a/c was higher in patients with monoclonal gammopathies than in those without (50% compared with 18%; P = 0.009). CONCLUSION: The prevalence of monoclonal gammopathies in patients with HCV related chronic liver disease is striking and is often associated with genotype 2a/c infection. PMID- 9729183 TI - Use of an algorithm for administering subcutaneous heparin in the treatment of deep venous thrombosis. AB - BACKGROUND: Despite the widespread use of subcutaneous heparin in the initial treatment of deep venous thrombosis, there are no guidelines for achieving adequate anticoagulation with this drug. OBJECTIVE: To implement a weight-based algorithm for the administration of subcutaneous unfractionated heparin after an intravenous loading dose. DESIGN: Prospective cohort study. SETTING: University hospital. PARTICIPANTS: 70 outpatients with proximal venous thrombosis. INTERVENTION: An intravenous bolus of heparin followed by a subcutaneous injection of heparin in doses adjusted for body weight. Subsequent adjustments of the subcutaneous heparin dose were scheduled twice daily according to the algorithm; the activated partial thromboplastin time (aPTT) was measured in the mid-interval (target range, 50 to 90 seconds). RESULTS: The therapeutic threshold aPTT (> or = 50 seconds) was achieved in 61 patients (87%) within 24 hours and in 69 patients (99%) within 48 hours. In 7 patients (10%), a supratherapeutic aPTT lasted more than 12 hours. No major bleeding episodes or cases of heparin-induced thrombocytopenia were seen. Three patients (4.3% [95% CI, 0.9% to 12.0%]) had recurrent thromboembolism during 3 months of follow-up. CONCLUSION: The administration of subcutaneous heparin according to a weight-based algorithm allows the rapid achievement of effective and safe anticoagulation in patients with deep venous thrombosis. PMID- 9729184 TI - The importance of supporting autonomy in medical education. AB - Many thoughtful leaders in medicine have asserted their belief that when physicians are more humanistic in their interactions with patients, their patients have more positive health outcomes. Consequently, many advocates have called for the practice of teaching students and residents to provide more humanistically oriented care. This article reviews research from motivational psychology, guided by self-determination theory, that suggests that when medical educators are more humanistic in their training of students, the students become more humanistic in their care of patients. Being humanistic in medical education can be achieved through support of the autonomy of students. Autonomy support means working from the students' perspectives to promote their active engagement and sense of volition with respect to learning. Research suggests that when educators are more supportive of student autonomy, students not only display a more humanistic orientation toward patients but also show greater conceptual understanding and better psychological adjustment. PMID- 9729185 TI - Update in gastroenterology. PMID- 9729186 TI - The ineffectiveness of immunosuppressive therapy in lymphocytic myocarditis: an overview. AB - BACKGROUND: The use of immunosuppressive therapy for myocarditis is controversial. PURPOSE: To review the literature on the effectiveness of immunosuppressive therapy in biopsy-proven lymphocytic myocarditis. DATA SOURCES: Two authors independently searched MEDLINE and other medical databases from 1980 to 26 June 1997. STUDY SELECTION: Randomized, controlled trials; matched-cohort studies; and case-control studies of patients with biopsy-proven myocarditis (Dallas criteria or a mean of > or = 2.5 lymphocytes per high-power field) for which any form of immunosuppressive treatment was used. The outcomes of interest were mortality and change in left ventricular ejection fraction. DATA EXTRACTION: 6 of 374 studies satisfied the selection criteria. DATA SYNTHESIS: In survivors, left ventricular function in myocarditis improved approximately 10% over 6 months without immunosuppressive treatment. Prednisone alone did not improve survival (P >0.2) or left ventricular function (P >0.11). Prednisone combined with azathioprine or cyclosporine did not improve survival (P >0.2) or left ventricular function (P >0.2) in three studies. However, one small matched-cohort study showed improvement in children (P <0.01). Neither interferon nor thymic hormone improved survival or left ventricular function. CONCLUSIONS: Immunosuppressive therapy is ineffective in lymphocytic myocarditis. Current therapy in lymphocytic myocarditis seems to be limited to supportive measures or transplantation. PMID- 9729187 TI - Treating histologically mild chronic hepatitis C: monotherapy, combination therapy, or tincture of time? AB - The recent National Institutes of Health Consensus Conference on hepatitis C solidified the justification for a selective approach to treatment. Nevertheless, the high profile of chronic hepatitis C has led to a sense of urgency about treating "all-comers" and thus has caused the variable natural history of this disease to be overlooked. The debate about whom to treat has failed to focus attention on the alternative approach of waiting for better emerging therapies for the subset of patients with histologically mild chronic hepatitis C. Practitioners should be more confident about postponing treatment in less symptomatic patients if liver biopsy specimens show no more than grade 1 necroinflammatory activity or stage 1 fibrosis. Patients with these lesions, in the absence of clinical signs of advancing disease, are much less likely than patients with higher grades or stages to progress to cirrhosis. A "cure" for chronic hepatitis C remains elusive. End points of treatment depend on the achievement of sustained clearance of serum hepatitis C virus RNA, which is influenced, in turn, by the patient's viral replication and immune balance. Treatment of histologically mild chronic hepatitis C may ultimately mimic that of HIV infection. PMID- 9729188 TI - New disease, old story. PMID- 9729189 TI - Antimicrobial chemotherapy for Legionnaires disease: time for a change. PMID- 9729191 TI - Letter to a patient's doctor. PMID- 9729190 TI - Chained smoker. PMID- 9729192 TI - Firearm injury prevention. PMID- 9729193 TI - Firearm injury prevention. PMID- 9729194 TI - Firearm injury prevention. PMID- 9729195 TI - Firearm injury prevention. PMID- 9729196 TI - Firearm injury prevention. PMID- 9729197 TI - Reframing gun violence. PMID- 9729198 TI - Reframing gun violence. PMID- 9729199 TI - Reframing gun violence. PMID- 9729200 TI - Reframing gun violence. PMID- 9729201 TI - Unusual involvement of bone in primary systemic amyloidosis. PMID- 9729203 TI - Secrecy in science: the flock worker's lung investigation. PMID- 9729202 TI - Recurrent spontaneous hepatic rupture in primary hepatic amyloidosis. PMID- 9729204 TI - Molecular Basis of Ion Channels and Receptors Involved in Nerve Excitation, Synaptic Transmission and Muscle Contraction. Proceedings of a conference in memory of Professor Shosaku Numa. Tokyo, Japan, January 12-15, 1993. PMID- 9729205 TI - Cancer Prevention: From the Laboratory to the Clinic: Implications of Genetic, Molecular, and Preventive Research. Proceedings of an international conference. New York City, New York, USA. September 22-24, 1994. PMID- 9729206 TI - Structure and Functions of the Human Prefrontal Cortex. Proceedings of a conference. New York City, New York, USA. March 2-4, 1995. PMID- 9729207 TI - Bone Marrow Transplantation: Foundations for the 21st Century. Proceedings of a conference. Orlando, Florida, USA. March 15-18, 1995. PMID- 9729208 TI - DNA Vaccines: a New Era in Vaccinology. Proceedings of a conference. Arlington, Virginia, USA. April 6-9, 1995. PMID- 9729209 TI - The Flight from Science and Reason. Proceedings of a conference. New York, New York, USA. May 31-June 2, 1995. PMID- 9729210 TI - Neuropeptides: Basic and Clinical Advances. Proceedings of the 1995 Summer Neuropeptide Conference. Martha's Vineyard, Massachusetts, USA. June 25-29, 1995. PMID- 9729211 TI - Recombinant DNA Biotechnology III: The Integration of Biological and Engineering Sciences. Proceedings of a conference. Deauville, France, October 16-21, 1994. PMID- 9729212 TI - Time-Dependent Structure and Control of Arterial Blood Pressure. Proceedings of a conference. Ferrara, Italy, September 10-12, 1995. PMID- 9729214 TI - Molecular and Developmental Biology of Certilage. Conference proceedings. Bethesda, Maryland, USA. September 27-30, 1995. PMID- 9729215 TI - Pharmacological Intervention in Aging and Age-Associated Disorders. Proceedings of the 6th Congress of the International Association of Biomedical Gerontology. Tokyo, Japan, August 20-23, 1995. PMID- 9729213 TI - Propedeutics to Cancer Management: Biology and Biochemistry of Normal and Cancer Cell Growth. Proceedings of a conference. Erice, Italy, April 1-6, 1995. PMID- 9729216 TI - Women and Mental Health. Proceedings of a conference. New York, New York, USA. March 10, 1995. PMID- 9729217 TI - Vector-Borne Pathogens: International Trade and Tropical Animal Diseases. Proceedings of a meeting. San Jose, Costa Rica, May 8-12, 1995. PMID- 9729218 TI - Engineering Plants for Commerical Products and Applications. Conference proceedings. Lexington, Kentucky, USA. October 1-4, 1995. PMID- 9729219 TI - Myocardial Preservation, Preconditioning, and Adaptation. Proceedings of a conference. Madras, India, December 14-16, 1995. PMID- 9729220 TI - Understanding Aggressive Behavior in Children. Proceedings of a conference. New York City, New York, USA. September 29-October 2, 1995. PMID- 9729221 TI - Cellular and Molecular Mechanisms of Drugs of Abuse: Cocaine, Ibogaine, and Substituted Amphetamines. Proceedings of a conference. Niigata City, Japan, June 29-30, 1995. PMID- 9729222 TI - Proceedings of the 2nd International Symposium on VIP, PACAP, and Related Peptides. New Orleans, Louisiana, USA. October 4-7, 1995. PMID- 9729223 TI - The Integrative Neurobiology of Affiliation. Proceedings of a conference. Washington, DC, USA. March 14-17, 1996. PMID- 9729224 TI - Atherosclerosis IV: Recent Advances in Atherosclerosis Research. Proceedings of the 4th Saratoga International Conference on Atherosclerosis. Hawaii, USA. February 6-8, 1996. PMID- 9729225 TI - Receptor Classification: The Integration of Operational, Structural, and Transductional Information. Proceedings of a conference. Verona, Italy, September 21-22, 1995. PMID- 9729226 TI - Proceedings of the 10th International Symposium on the Pharmacology of Thermoregulation. Memphis, Tennessee, USA. August 17-22, 1996. PMID- 9729227 TI - Neuropeptides in Development and Aging. Proceedings of a conference. Breckenridge, Colorado, USA. February 3-6, 1996. PMID- 9729228 TI - Adolescent Gynecology and Endocrinology: Basic and Clinical Aspects. Proceedings of the 3rd International Congress. Athens, Greece, December 6-9, 1995. PMID- 9729229 TI - Psychobiology of Postraumatic Stress Disorder. Proceedings of a conference. New York, New York, USA. September 7-10, 1996. PMID- 9729230 TI - Proceedings of the 3rd International Conference on Neuroprotective Agents: Clinical and Experimental Aspects. Lake Como, Italy, September 9-12, 1996. PMID- 9729231 TI - Proceedings of the VIIIth International Conference on the Na/K-ATPase and Related Transport ATPases. Mar del Plata, Argentina, August 26-30, 1996. PMID- 9729232 TI - Enhancement of cytotoxicity and clastogenicity of l-DOPA and dopamine by manganese and copper. AB - It is an increasingly popular hypothesis that the continued degeneration of dopaminergic neurons in Parkinson's disease (PD) may be the consequence of aberrant oxidation of dopamine and resultant generation of DNA reactive species in PD patients receiving levodopa (l-DOPA) therapy. Occupational metal exposure, particularly to manganese, is a risk factor for Parkinsonism and manganese has been shown to be a true catalyst for dopamine oxidation lending support to this hypothesis. In the present studies, we demonstrate that the antiproliferative activity of l-DOPA and dopamine on Chinese Hamster V79 cells is enhanced by at least an order of magnitude by concomitant exposure to manganese chloride or copper sulfate (500 microM), but not to iron(III) or zinc. Moreover, manganese and copper confer strong clastogenic properties to both compounds as detected in an in vitro micronucleus assay in V79 cells. Metal catalyzed oxidation of drug was associated with the development of a brown-black particulate substance presumed to be a melanin precursor formation. The extent of formation of this precursor paralleled clastogenicity. Metal-enhanced effects were completely antagonized by the concurrent addition of cysteine or reduced glutathione to the cultures. These findings are in support of the hypothesis that aberrant oxidation of dopamine resulting from non-physiological levels of catalytic metals may contribute to the death of dopaminergic neurons and further suggest that oxidation-dependent DNA damage may be the basis for this cell death. PMID- 9729233 TI - Differential expression of the two forms of prolactin receptor mRNA within microdissected hypothalamic nuclei of the rat. AB - The prolactin receptor (PRL-R) has recently been identified in various hypothalamic nuclei of female rats. In this study, expression of both the short- and long-forms of PRL-R mRNA was investigated in 11 microdissected hypothalamic nuclei of ovariectomized, estrogen-treated rats. Specific nuclei were micropunched from 300-micrometer thick frozen coronal sections with autoclaved stainless steel needles of 300 or 500 micrometer diameter. Total RNA was extracted from the punched tissue, and the two forms of PRL-R mRNA were detected by reverse transcription polymerase chain reaction (RT-PCR) using specific primers. The RT-PCR product was verified by Southern hybridization with a digoxigenin-labelled oligonucleotide probe common to both forms. The results showed that both forms of PRL-R mRNA were expressed to varying degrees in the choroid plexus, cerebral cortex and various hypothalamic nuclei, including: ventromedial preoptic nucleus, ventrolateral preoptic nucleus, medial preoptic nucleus, suprachiasmatic nucleus, supraoptic nucleus, paraventricular hypothalamic nucleus, periventricular hypothalamic nucleus, arcuate nucleus, ventromedial hypothalamic nucleus, and median eminence. Of these brain regions, the choroid plexus expressed the highest level while the suprachiasmatic nucleus contained the lowest level of mRNA. There was no expression detected in the dorsomedial hypothalamic nucleus. The choroid plexus, supraoptic nucleus and paraventricular hypothalamic nucleus had higher levels of the short-form of the PRL-R mRNA than the long-form, whilst other hypothalamic nuclei preferentially expressed the long-form of the PRL-R mRNA. The differential expression of PRL-R gene suggests that the two forms may be differentially regulated in specific brain regions and may mediate different functions of PRL. PMID- 9729234 TI - Increased inducible nitric oxide synthase protein but limited nitric oxide formation occurs in astrocytes of the hph-1 (tetrahydrobiopterin deficient) mouse. AB - It has been suggested that decreased tetrahydrobiopterin (BH4) availability may be a useful tool for limiting excessive nitric oxide (NO) formation. In order to test this hypothesis we utilised cultured astrocytes derived from the brain of the hph-1 (BH4 deficient) mouse. In response to treatment with lipopolysaccharide and interferon-gamma (LPS/gammaIFN) levels of BH4 doubled in both wild type and hph-1 astrocytes. However, levels of BH4 in hph-1 astrocytes remained only 25% of the wild type astrocytes. Nitric oxide formation, measured with an NO-electrode, was 45% less from LPS/gammaIFN stimulated hph-1 astrocytes compared with wild type stimulated astrocytes. In contrast, iNOS specific activity and iNOS protein were enhanced in hph-1 stimulated astrocytes by 40 and 60%, respectively when compared with wild type. In conclusion it appears that whilst a decrease in BH4 may limit NO release per se, the possibility and consequences of long term 'over' induction of iNOS protein requires further consideration. PMID- 9729235 TI - Follow-up of interhemispheric differences of motor evoked potentials from the 'affected' and 'unaffected' hemispheres in human stroke. AB - We analysed motor-evoked potentials (MEPs) from the hand muscles during focal transcranial magnetic stimulation (TCS) in both the affected (AH) and the unaffected (UH) hemispheres of 17 monohemispheric stroke patients followed-up in subacute stage. Recording sessions were performed at 2 (T1 session) and 4 (T2 session) months from acute stroke. Clinical and functional scores were evaluated. An age-sex matched group of 20 healthy subjects have been referenced. In T1, relaxed MEPs from AH were smaller than UH (p<0.001) and normals (p<0.001). In T2, an increase of AH relaxed-MEPs amplitude was observed, combined with an improvement of clinical and functional scores (p<0.001). On the other hand, the amplitude of contracted MEPs from the AH in T1 was larger than in the normal group. This parameter decreased toward normal limits in T2, provided that the amplitude of the MEPs from the AH improved, while it further increased when TCS of the AH continued to fail in eliciting MEPs. This phenomenon was statistically combined with clinical improvement of disability and neurological scores. Recovery of the excitability AH threshold with progressive 'balancing' of the UH hyperresponsiveness represents a good prognostic parameter for clinical outcome of hand motor function. PMID- 9729236 TI - Cellular localization of tumor necrosis factor alpha following focal cerebral ischemia in mice. AB - Tumor necrosis factor alpha (TNFalpha) is a pleiotrophic cytokine with diverse proinflammatory actions. Focal cerebral ischemia induces rapid and dramatic increases in TNFalpha levels within and surrounding the focus of damaged brain both in striatum and cortex. The actions of TNFalpha during cerebral ischemia may be related to the cell types which deliver and/or accept TNFalpha signals. However, the cellular sources of TNFalpha following cerebral ischemia have not been fully elucidated. The present study was designed to determine the cellular localization of TNFalpha following permanent middle cerebral artery occlusion (MCAO) in mice. As judged by immunohistochemistry, TNFalpha expression in the ischemic hemisphere was increased at 3 h following MCAO, peaked at 6 to 12 h, and decreased at 24 h. Double immunostaining for TNFalpha and neuron specific enolase (NSE) or glial fibrillary acidic protein (GFAP) showed that TNFalpha positive neurons were observed in both the ischemic core and perifocal region, while TNFalpha positive astrocytes were observed in the outer cortical layer, the corpus callosum, the molecular layer of the hippocampus, and periventricular areas. The presence of TNFalpha immunoreactivity in neurons and nerve fibers following MCAO suggests that TNFalpha expressed in ischemic neurons might be delivered via axonal transport, while TNFalpha immunoreactivity in astrocyte end feet and ependymal cells following MCAO suggests that TNFalpha may be involved in blood-brain barrier disruption and the initiation of inflammation in the brain. PMID- 9729237 TI - Effect of ex vivo hyperthermia on radiation-induced micronuclei in lymphocytes of cancer patients before and during radiotherapy. AB - To investigate the effect of ex vivo hyperthermia (HT) and 137Cs-irradiation on micronucleus (MN) production in cytokinesis-blocked lymphocytes, we obtained the peripheral blood samples from the same cancer patients (n=6) before and during fractionated partial-body radiotherapy (xRT). The whole blood cultures were heated at 43.5 degrees C for 60 min, followed by 137Cs irradiation (0-4 Gy). The control cultures from the same patients were incubated at 37 degreesC after being exposed to radiation. The lymphocytes were then stimulated with PHA. Cytochalasin B was applied at 44 h, and lymphocytes were harvested at 72 h. MN frequency was determined on Giemsa-stained slides. We found that in patients before xRT, HT (43.5 degrees C) significantly increased the MN yield (mean+/-SEM) in unirradiated lymphocytes from 15.6+/-2.8 (37 degrees C) to 39.7+/-10.9. Further, in patients either before or during xRT, when the lymphocytes were treated with HT (43.5 degrees C) and combined with ex vivo irradiation, the MN yield (Y) could be estimated by a linear equation Y=C+alphaD. Our findings indicate that as measured by the MN production in cytokinesis-blocked lymphocytes, HT alone at 43.5 degrees C++ induced DNA damage. Moreover, it enhanced the radiation-induced cytogenetic damage. Therefore, the application of HT may impair the T-cell function in cancer patients who are receiving radiotherapy. 1998 Elsevier Science B.V. PMID- 9729238 TI - Fine structural analysis of DNA repair in mammalian cells. AB - Nucleotide excision repair (NER) of ultraviolet (UV) light induced photo lesions is heterogeneous in the genomic DNA. We have investigated the mechanistic basis for this repair heterogeneity by analyzing NER activity in higher order chromatin of repair proficient hamster cells. Immunological labeling of repair and transcription sites indicates that NER initiates at the nuclear matrix in close association with transcription. The repair gradually extends into the loop DNA regions in a time dependent fashion. Repair analysis indicates that the DNA damaged by UV irradiation is recruited to the nuclear matrix soon after UV exposure. Consistent with this finding, immunofluorescence and western blotting analyses indicate the enrichment of many NER proteins (XPA, RPA, PCNA, the P62 and p89 sub-units of the basal transcription factor, TFIIH) in the nuclear matrix of UV treated cells. These results strengthen the notion that the nuclear matrix is an important site for the assembly of an efficient repair complex. PMID- 9729239 TI - Hypothalamic galanin: control by signals of fat metabolism. AB - The peptide, galanin (GAL), is known to stimulate eating behavior, reduce energy expenditure and affect the release of metabolic hormones. Further, the activity of this peptide in the hypothalamus is modulated, in turn, by these hormones as well as by the ingestion of nutrients. The focus of this investigation is on signals related to nutrient metabolism that may also affect GAL production and, through these neurochemical events, control the ingestion of specific nutrients. Three experiments were performed in normal-weight male, Sprague-Dawley rats. In Experiment 1, the impact of food deprivation (24 and 48 h) was examined. Experiment 2 tested the effects of the compound, 2-deoxy-D-glucose (2-DG, 200 and 400 mg/kg), which blocks glucose utilization, whereas Experiment 3 studied mercaptoacetate (MA, 200 and 600 micromol/kg), which blocks fatty acid oxidation. Eating behavior was examined in some rats, whereas hypothalamic GAL activity was measured in others using radioimmunoassay, immunohistochemistry and in situ hybridization. Both food deprivation and MA (600 micromol/kg), but not 2-DG, affected GAL in the hypothalamus, in one specific area. This is the anterior parvocellular region of the paraventricular nucleus (aPVN), which has a dense concentration of GAL-containing neurons and terminals. GAL gene expression and peptide immunoreactivity in this area is enhanced by food deprivation; in contrast, it is reduced by injection of MA. Other hypothalamic sites with dense concentrations of GAL-containing neurons or fibers are unaffected by food deprivation or MA, and the antimetabolite 2-DG has no impact on GAL in any area. Behavioral measurements indicate that these shifts in GAL activity are accompanied by specific changes in eating behavior. Food deprivation which enhances aPVN GAL produces a marked increase in fat ingestion, whereas MA which reduces aPVN GAL causes a specific reduction in fat ingestion along with a stimulation of protein intake. In contrast, 2-DG preferentially enhances ingestion of carbohydrate. These findings suggest a possible relationship between GAL activity in the aPVN and the metabolic and behavioral processes of fat metabolism and ingestion. PMID- 9729240 TI - Identification of structural features and associated mechanisms of action for carcinogens in rats. AB - A set of chemicals tested for carcinogenicity in rats that have been analyzed in the Carcinogenic Potency Database (CPDB) was subjected to CASE/MULTICASE (a computer-automated structure evaluation system) structure-activity relationship (SAR) analyses. This SAR system identifies structural features of chemicals in a learning set that are associated with a predefined activity and produces an SAR model based on these characteristics. The rat CPDB used in this study consisted of 745 chemicals, 383 of which are carcinogens, 14 marginally active carcinogens (i.e., chemicals that require a relatively high dose to induce carcinogenesis) and 348 are non-carcinogens. In an internal prediction analysis where CASE/MULTICASE 'predicted' the activity of chemicals in the learning set, the system was able to achieve a concordance between experimental and predicted results of 95%. This indicates that the program is able to adequately assess the chemicals in the database. In a 10-fold cross-validation study where 10 disjoint sets of 10% of the chemicals were removed from the database and the remaining 90% of the chemicals were used as a learning set, CASE/MULTICASE was able to achieve a concordance between experimental and predicted results of 64%. Using a modified validation process designed to investigate the predictivity of a more focused SAR model, the system was able to achieve a concordance of 71% between experimental and predicted results. Among the major biophores identified by CASE/MULTICASE as associated with cancer causation in rats, several are derived from electrophilic or potentially electrophilic compounds (e.g., aromatic amines, nitrogen mustards, isocyanates, epoxides). Other biophores however are derived from chemicals seemingly devoid of actual or potential DNA-reactivity and as such may represent structural features of non-genotoxic carcinogens. PMID- 9729241 TI - Cytogenetic monitoring in coke oven workers. AB - Cytogenetic markers (chromosomal aberrations, sister chromatid exchanges (SCE), cells with high frequency of SCE (HFC), the heterogeneity index SCE (SCE-H) and genetic polymorphism of genotypes GSTM1 and NAT2 were evaluated in the peripheral lymphocytes of 64 coke oven workers and 34 control subjects from the same plant. Personal monitors were used to evaluate exposure to eight carcinogenic (polycyclic aromatic hydrocarbons) PAHs, including B[a]P, during an 8-h working shift. Smoking habits were checked by urinary cotinine measurement. The exposure among coke oven workers ranged widely from 0.6 to 547 microgram/m3 and 2 to 50 137 ng/m3, for carcinogenic PAHs and B[a]P, respectively. The respective values in controls were 0.07 to 1.51 microgram/m3 and from 2 to 63 ng/m3. The results of biomonitoring in exposed vs. control subjects were as follows: frequency of chromosomal aberrations (% AB.C.), 2. 30% AB.C. vs. 1.09% AB.C. (P<0.05); sister chromatid exchanges, 7.47 SCE/cell vs. 5.49 SCE/cell (P<0.05); HFC, 5.94% vs. 2.06% (P<0.05) and SCE-H index, 1.49 vs. 1.01 (P<0.05). All the cytogenetic markers were significantly increased in the exposed vs. control groups. The effect of smoking was observed only in SCE when evaluated as HFC. Using individual exposure data for carcinogenic PAHs, a significant correlation between exposure and %AB.C. (r=0.372, P=0.0002), SCE/cell (r=0.331, P=0.001), HFC (r=0.467, P=0.007) and SCE/H (r=0. 286, P=0.004) was found. No effects of GSTM1 and NAT2 genotypes, individually or in combination, on the cytogenetic markers was observed. It is concluded that occupational exposure of coke oven workers involved in this study resulted in an increased level of chromosomal aberrations and SCE. The frequency of AB.C. and SCE/cell was found to be related to exposure to carcinogenic PAHs. PMID- 9729242 TI - Latexin expression in smaller diameter primary sensory neurons in the rat. AB - Most of the smaller diameter neurons of dorsal root and trigeminal ganglia in adult rats expressed latexin, which has the inhibitor activity of carboxypeptidase A. Most of the dorsal root ganglion (DRG) neurons containing either calcitonin gene-related peptide (CGRP), substance P (SP) or somatostatin (SST) coexpressed latexin. Latexin was widely distributed in the cytoplasm of the cell body and in axonal fibers of cultured DRG neurons which were sensitive to capsaicin. In addition, latexin-immunoreactivity was observed throughout lamina II of the spinal cord in normal animals, but was lost following sciatic nerve axotomy, suggesting the presence of latexin-immunoreactive axonal fibers and/or terminals from DRG neurons. Immunoelectron microscopy indeed revealed latexin immunoreactive axonal terminals and thinly myelinated and unmyelinated axonal fibers within the dorsal horn. These observations suggest that latexin may be involved in nociceptive information transmission or its modulation. PMID- 9729244 TI - No association between Alzheimer plaques and decreased levels of cytochrome oxidase subunit mRNA, a marker of neuronal energy metabolism. AB - It has been proposed that neuritic plaques or toxic substances diffusing from them contribute to neurodegeneration in Alzheimer disease. We examined this hypothesis by looking for evidence of decreased neuronal energy metabolism in the proximity of neuritic plaques. Levels of mitochondrial DNA-encoded mRNA for subunit III of cytochrome oxidase, a marker of neuronal energy metabolism, were determined in post mortem brain samples. Consistent with earlier results, overall cytochrome oxidase subunit III mRNA levels were decreased in Alzheimer midtemporal cortex compared with controls. However, this reduction did not correlate with plaque density. In Alzheimer brains, cytochrome oxidase subunit III mRNA levels in neurons bearing neurofibrillary tangles were lower than in tangle-free neurons. However, neuronal cell bodies in close proximity of neuritic plaques showed no decrease in cytochrome oxidase subunit III mRNA or total polyadenylated mRNA compared with more distant neurons. Cytochrome oxidase enzyme activity in neuronal processes also showed no local reduction around neuritic plaques. These results suggest that neuritic plaques do not contribute to reduced neuronal energy metabolism in Alzheimer disease. PMID- 9729243 TI - Selective changes in AMPA receptors in rabbit cerebellum following classical conditioning of the eyelid-nictitating membrane response. AB - alpha-Amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) receptors are critically involved in several forms of synaptic plasticity proposed to be neural substrates for learning and memory, e.g., long-term potentiation and long-term depression (LTD). The present study was designed to determine changes in cerebellar AMPA receptors following classical conditioning of the eyeblink nictitating membrane response (NMR) in the rabbit. Quantitative autoradiography was used to assess changes in ligand binding properties of cerebellar AMPA receptors following NMR conditioning elicited by pairing electrical stimulation of the pontine nuclei with an airpuff to the eye. [3H]AMPA and [3H]-6-cyano-7 nitroquinoxaline-2,3-dion (CNQX) binding were determined following preincubation of frozen-thawed brain tissue sections at 0 or 35 degreesC. With 0 degreesC preincubation, no significant differences in [3H]AMPA binding to cerebellar AMPA receptors were seen between any of the experimental groups tested. In contrast, preincubation at 35 degreesC revealed significant decreases in [3H]AMPA binding to the trained side of the cerebellar cortex resulting from paired presentations of the conditioned and the unconditioned stimuli, while unpaired presentations of the stimuli resulted in no significant effect. With 35 degreesC preincubation, there were no significant differences in [3H]CNQX binding between any of the experimental groups and no significant differences in [3H]AMPA binding in the untrained side of the cerebellum. These results indicate that NMR conditioning is associated with a selective modification of AMPA-receptor properties in brain structures involved in the storage of the associative memory. Furthermore, they support the hypothesis that cerebellar LTD, resulting from decreased synaptic efficacy at parallel fiber-Purkinje cell synapses mediated by a change in AMPA receptor properties, is a form of synaptic plasticity that supports this type of learning. PMID- 9729245 TI - The underlying structure of the subtelomeric region detected by microphotometrical scanning and chromosome graphic image analysis. AB - Previous research showed that the microphotometrical scanning of T-banded subtelomeric regions reveals the presence of specific patterns of the Giemsa stain density distributions as detected in chromosomes of normal human lymphocytes and CHO cells. Analyses with this method of the T-banded subtelomeric segments of CHO endoreduplicated chromosomes confirmed that these density patterns replicate in a similar way in sister chromosomes. Besides, the specific removal of portions of the subtelomeric segments appearing as tiny holes located where these density patterns are found suggested that both phenomena are related. The possible connection of these findings to the molecular structure of the subtelomeric region is briefly discussed. PMID- 9729246 TI - Localization of chromosome breakpoints: implication of the chromatin structure and nuclear architecture. AB - Restriction endonucleases and ionizing radiations have been extensively used to study the origin of chromosomal aberrations. Although a non-random distribution of chromosome breakpoints induced by these agents has been claimed by several authors, the significance of the chromatin structure and nuclear architecture in the localization of breakpoints is still not well understood. Breakpoint patterns produced by endonucleases targeted to specific genome sequences or by ionizing radiations could provide additional evidence to clarify this point. Results obtained from the localization of breakpoints induced by AluI, BamHI or DNase I as well as by neutrons or gamma-rays in G-banded Chinese hamster ovary (CHO) chromosomes are presented. AluI and BamHI were electroporated into CHO cells either during the G1 or S-phase of the cell cycle. A co-localization of breakpoints was found with a preferential occurrence in G-light bands independent of the cell cycle stage in which aberration production took place. Since AluI and BamHI recognition sequences are partitioned in the housekeeping and tissue specific subgenomes respectively, we postulated that nuclease sensitive sites in active chromatin could be the main targets for the induction of breakpoints by these endonucleases. This assumption is supported by the finding that DNase I induced breakpoint patterns in CHO cells are similar to those produced by AluI and BamHI. Digestion of fixed CHO chromosomes with these endonucleases induced G banding suggesting a higher sensitivity of G-light chromatin. For comparison purposes, CHO cells were irradiated with neutrons or gamma-rays and breakpoints localized in G-banded chromosome aberrations. A higher occurrence of breakpoints in G-light bands was also observed. We detected seven breakage-prone G-light bands that were preferentially damaged by the three endonucleases and by both types of radiation. These results emphasize the possible implication of the chromatin structure and the nuclear architecture in the localization of chromosome breakpoints induced by endonucleases, neutrons and gamma-rays. PMID- 9729247 TI - Recommendations for statistical designs of in vivo mutagenicity tests with regard to subsequent statistical analysis. AB - A workshop was held on September 13 and 14, 1993, at the GSF, Neuherberg, Germany, to start a discussion of experimental design and statistical analysis issues for three in vivo mutagenicity test systems, the micronucleus test in mouse bone marrow/peripheral blood, the chromosomal aberration tests in mouse bone marrow/differentiating spermatogonia, and the mouse dominant lethal test. The discussion has now come to conclusions which we would like to make generally known. Rather than dwell upon specific statistical tests which could be used for data analysis, serious consideration was given to test design. However, the test design, its power of detecting a given increase of adverse effects and the test statistics are interrelated. Detailed analyses of historical negative control data led to important recommendations for each test system. Concerning the statistical sensitivity parameters, a type I error of 0.05 (one tailed), a type II error of 0.20 and a dose related increase of twice the background (negative control) frequencies were generally adopted. It was recommended that sufficient observations (cells, implants) be planned for each analysis unit (animal) so that at least one adverse outcome (micronucleus, aberrant cell, dead implant) would likely be observed. The treated animal was the smallest unit of analysis allowed. On the basis of these general consideration the sample size was determined for each of the three assays. A minimum of 2000 immature erythrocytes/animal should be scored for micronuclei from each of at least 4 animals in each comparison group in the micronucleus assays. A minimum of 200 cells should be scored for chromosomal aberrations from each of at least 5 animals in each comparison group in the aberration assays. In the dominant lethal test, a minimum of 400 implants (40-50 pregnant females) are required per dose group for each mating period. The analysis unit for the dominant lethal test would be the treated male unless the background frequency of dead implants (DI) is so low that multiple males would need to be integrated to meet the minimum observation of one adverse outcome (DI) per analysis unit. A three-step strategy of data analysis was proposed for the cytogenetic assays. Use of negative historical controls was allowed in certain circumstances for interpretation of results from micronucleus tests and chromosomal aberration tests. PMID- 9729248 TI - Epileptiform activity and changes in field potential responses induced by low [Mg2+]0 in a genetic rat model of absence epilepsy. AB - The genetic absence epilepsy rats of Strasbourg (GAERS) display alterations in cortical synaptic transmission possibly facilitating the generation of ictaform activity and the late development into convulsive epilepsy. We studied low Mg2+ induced epileptiform activities and their long term effects on field potentials (fp) evoked by paired pulse stimulation in hippocampal area CA1 (CA1), medial entorhinal cortex (EC) and frontal cortex (FC) in in-vitro-slice preparations from GAERS and control (NE) adult rats (6 months). Omitting Mg2+-ions from artificial cerebrospinal fluid (ACSF) caused recurrent short discharges (in CA1) and seizure-like events (in EC) in both GAERS and NE rats. Latency to onset of activity as well as discharge pattern, frequency and amplitude of such events did not differ between the two strains, neither in CA1 nor in EC. In the FC, however, epileptiform events occurred in NE rats, but not in GAERS. Field potentials in normal ACSF were similar in both strains in CA1 and FC, while they were smaller in the EC of GAERS. Low [Mg2+]0 caused long-term changes of fp only in area CA1 where the population spikes were depressed in GAERS and increased in NE rats. We concluded that susceptibility to low [Mg2+]0-induced epileptic activity in EC and hippocampal area CA1 is not higher in GAERS than in NE adult rats. However, some properties like synaptic coupling in EC and long-term changes in synaptic efficacy induced by epileptiform activity in CA1 differ from that in NE rats. Whether the particularities in GAERS may be related to kindling by absence epileptic activities will be studied in further experiments. PMID- 9729249 TI - Site and behavioral specificity of periaqueductal gray lesions on postpartum sexual, maternal, and aggressive behaviors in rats. AB - Bilateral electrolytic lesions of the lateral and ventrolateral caudal periaqueductal gray (cPAGl,vl) of lactating rats are known to severely reduce suckling-induced kyphosis (upright crouched nursing), which is necessary for maximal litter weight gains, and impair sexual behavior during the postpartum estrous, while heightening nursing in other postures and attacks on unfamiliar adult male intruders. In the present report, the site specificity of the cPAG with respect to the control of these behaviors was determined by comparing lesions of the cPAGl,vl with similarly sized lesions within the rostral PAG (rPAG) and surrounding mesencephalon. The previously seen effects of prepartum cPAGl,vl lesions on kyphotic nursing, sexual proceptivity and receptivity, maternal aggression, and daily litter weight gains were replicated. Additionally, the post-lesion facilitation of aggression was found to be behaviorally specific, first by being directed toward an adult, but not to a nonthreatening juvenile male rat, and second, by requiring the recent presence of the pups, being eliminated or decreased 24 h after removal of the litter. Damage to the rPAG did not affect nursing or sexual behaviors, and had only a minimal effect on maternal aggression. Lesions of the rPAG, however, greatly impaired the dams' ability to rapidly release pups held in the mouth, but not to pick them up or carry them directly to the nest during retrieval. Separate regions of the PAG, therefore, are differentially involved in the control of specific components of behaviors in lactating rats. PMID- 9729250 TI - Dorsomedial hindbrain participation in glucoprivic feeding response to 2DG but not 2DG-induced hyperglycemia or activation of the HPA axis. AB - 2-Deoxy-d-glucose (2DG) is a glucose analogue that inhibits intracellular utilization of glucose and produces a characteristic behavioral response known as glucoprivic feeding. The area postrema (AP) is a caudal hindbrain structure shown previously to be involved in 2DG-induced glucoprivic feeding. In addition, peripheral administration of 2DG is known to elicit activation of both the hypothalamic-pituitary-adrenal (HPA) axis and the sympathoadrenomedullary system. The neural substrates for these neuroendocrine and neural responses to 2DG are not known although they may also involve the AP. The possible role of the AP in 2DG-induced feeding, activation of the HPA axis and hyperglycemia was investigated in Sprague-Dawley rats with lesions centered on the area postrema (APX) and sham-operated (SHM) rats administered 2DG (200 mg/kg) or physiological saline (1 ml/kg). Peripheral administration of 2DG evoked a feeding response in SHM rats that was abolished in APX animals. Interestingly, 2DG administered at this dose produced a significant increase in plasma corticosterone and plasma glucose in both SHM and APX rats for up to 4 h after drug treatment. Collectively, these findings suggest that the AP is involved in the behavioral (feeding) response to peripheral administration of 2DG, but does not appear to be a common neural substrate for the neuroendocrine (HPA axis) and sympathoadrenal (hyperglycemic) responses to this agent. PMID- 9729251 TI - Steady-state properties of sodium channels from healthy and tumorous human brain. AB - This extensive bilayer study of unpurified human brain channels from non-diseased and tumorous human brain involves more than 300 lipid bilayer experiments. Single channel conductances and subconductances, single channel fractional open times, the voltage-dependence of tetrodotoxin (TTX) block and the steady-state activation behavior of four different human brain synaptosomal preparations have been examined. Reproducible values have been obtained for the molecular electrophysiological parameters and their standard deviations, providing a database for future comparisons involving disease or drug-related changes in molecular sodium channel functions. In comparison with sodium channels from other species and under other experimental conditions, the bilayer system proved to be a reliable experimental setting. Despite the very different histology of the tissue probes, there were no significant differences in any of the examined electrophysiological features. PMID- 9729252 TI - Reciprocal changes in hypothalamic receptor binding and circulating leptin in anorectic tumor-bearing rats. AB - Although reduced biological activity of the obese gene product, leptin, has been associated with obesity, little information is available concerning leptin alterations during anorexia. Therefore, we measured circulating leptin concentrations and hypothalamic leptin binding in anorectic tumor-bearing and pair-fed control rats. Plasma concentrations of leptin decreased in tumor-bearing rats early in the course of tumor growth, and fell to nearly non-detectable levels during severe anorexia. The pair-fed control rats that ate the same amount of food as did the anorectic tumor-bearing rats exhibited a 50% decrease in plasma leptin concentration. Concentrations of free fatty acids were elevated in both tumor-bearing and pair-fed groups, while circulating levels of triglycerides were increased only in anorectic tumor-bearing rats. Leptin receptor density was doubled in the hypothalamus of tumor bearing rats, while binding affinity was decreased by 50%. These results suggest that peripheral leptin production is down regulated, perhaps due to increased lipolysis in tumor-bearing rats. It appears that hypothalamic leptin systems up-regulate receptor numbers in response to decreased blood leptin level, however, the decrease in binding affinity may compensate for these alterations. Therefore, the influence of leptin on hypothalamic neuropeptide Y feeding systems may be minimal in anorectic tumor bearing rats. PMID- 9729253 TI - Some aspects on radiation induced transmissible genomic instability. AB - The early observations on the possible induction of transmissible genomic instability after exposure to ionising radiation has received a strong support in the literature during the last 10 years. Aided by new research tools in biology, the better understanding of the mechanisms behind genomic instability leads to conclusions which are challenging the existing views on the interaction and response of the genome to radiation or chemicals. It has become commonly accepted that the full revelation of biological pathways leading to the loss of stability of the genome will also be a major step in the understanding of carcinogenesis. In this short review, some aspects of the recent knowledge and their implications are discussed. PMID- 9729254 TI - Salt appetite: interaction of forebrain angiotensinergic and hindbrain serotonergic mechanisms. AB - Methysergide injected bilaterally into the lateral parabrachial nucleus (LPBN) increases NaCl intake in several models of renin-dependent salt appetite. The present study investigated the role of angiotensin Type 1 (AT1) receptors in the subfornical organ (SFO) on this effect. The intake of 0.3 M NaCl and water was induced by combined administration of the diuretic, furosemide (FURO), and the angiotensin-converting enzyme inhibitor, captopril (CAP). Pretreatment of the SFO with an AT1 receptor antagonist, losartan (1 microgram/200 nl), reduced water intake but not 0.3 M NaCl intake induced by subcutaneous FURO+CAP. Methysergide (4 microgram/200 nl) injected bilaterally into the LPBN increased 0.3 M NaCl intake after FURO+CAP. Losartan injected into the SFO prevented the additional 0. 3 M NaCl intake caused by LPBN methysergide injections. These results indicate that AT1 receptors located in the SFO may have a role in mediating an enhanced sodium intake produced by methysergide treatment. PMID- 9729255 TI - The alkaline single cell gel electrophoresis: a new test for assessing DNA single strand breaks in Neurospora crassa. AB - The single cell gel electrophoresis (comet assay) is a potent technique in testing double and single strand breaks in DNA. In this paper, we present an application of alkaline comet assay to filamentous fungi for genotoxicological assessment of heavy metals for the first time. A wild strain of Neurospora crassa SLA 4200 was grown in presence of cadmium sulfate (CdSO4) (10 microM and 100 microM) for 12 h. Protoplasts from 12-h old mycelia were prepared by using Novozym 234 and DNA damage was evaluated by alkaline comet assay. Hydrogen peroxide (H2O2) (50 microM and 100 microM) was taken as an internal standard for DNA damage. Both CdSO4 and H2O2 induced significant single strand breaks in DNA. The results indicate that alkaline comet assay is a sensitive and rapid method for DNA damage analysis in filamentous fungal systems. PMID- 9729256 TI - Genotoxic activity of chlorinated butenoic acids in Salmonella typhimurium strains TA98, TA100 and TA104. AB - The mutagenic activities of several chlorinated butenoic acids, recently identified in chlorinated drinking waters, were determined by the Salmonella microsome assay. The Salmonella typhimurium tester strains TA98, TA100, and TA104 were used without S9 mix. The results from the investigation showed that (Z)-2 chloro-3-(dichloromethyl)-4-oxobutenoic acid (MX, in the open form) was the most potent mutagen of the compounds tested. However, a significant number of mutations was also induced by compounds with structural similarities to MX. In general, all the compounds, except the butenedioic acids, were mutagenic in the assays for both base-pair substitution strains (TA100, TA104) and for the frameshift strain TA98, with the highest mutagenic response observed in strain TA100. When the aldehyde group of MX and of 2-chloro-3-(chloromethyl)-4 oxobutenoic acid (CMCF, in the open form) was replaced by a dichloromethyl group, the mutagenic response in strains TA98 and TA104 changed. We concluded that a frame-shift mutation occurred because of the replacement. The increase of the TA104 mutagenicity suggested that adenosine could be the target for these types of compounds. Further evidence for such possibility were the modified adenosine adducts we could identify for some chlorinated butenoic acids. PMID- 9729257 TI - Mycoplasma fermentans glycolipid triggers inflammatory response in rat astrocytes. AB - Mycoplasma fermentans glycolipid (MfGL-II) is a major lipid in the membranes of this AIDS-associated mycoplasma and constituting up to 20% of the total phospholipids of this organism. It was recently shown that MfGL-II, mainly through its phosphocholine moiety, is responsible for the attachment of M. fermentans to host cells. We now show that MfGL-II is also associated with the secretion of inflammatory mediators by cells of the central nervous system. Stimulation of primary rat astrocytes by MfGL-II caused activation of protein kinase C, secretion of nitric oxide (NO) and prostaglandin E2, and augmented glucose utilization and lactate formation in a dose-dependent manner. In an attempt to define the minimal structural requirements for MfGL-II activity, the two O-acylated fatty acids in the molecule were removed. Deacylation pronouncedly reduced the stimulatory activity of the glycolipid, suggesting that the fatty acyl residues are essential. Incubation of MfGL-II with polyclonal anti-MfGL-II antiserum or with monoclonal anti-phosphocholine antibody diminished NO release, whereas incubation of MfGL-II with normal rabbit serum had no effect. It is, therefore, likely that the terminal phosphocholine moiety plays an important role in MfGL-IIs stimulation of glial cells. PMID- 9729258 TI - Effects of an inhibitor of topoisomerase II, ICRF-193 on the formation of ultraviolet-induced chromosomal aberrations. AB - Treatments of Chinese hamster V79 cells during one cell cycle with a new type of topoisomerase II inhibitor, ICRF-193, which does not accumulate cleavable topoisomerase-DNA complexes induced both chromosome- and chromatid-type aberrations with high frequencies. Furthermore, ICRF-193 synergistically enhanced the yield of UVB-induced chromatid-type aberrations, chromatid exchanges in particular. Treated with ICRF-193 for the last 3 h before harvest, cells showed frequent incidence of chromatid-type aberrations and synergistic enhancement of UVB-induced chromatid-type aberrations, chromatid exchanges in particular. These results suggest that spontaneous and UVB-induced lesions might be ultimately transformed into chromatid-type aberrations by topoisomerase II-dependent checkpoint process(es) in the G2 phase of the cell cycle. PMID- 9729259 TI - Effect of chronic restraint stress and tianeptine on growth factors, growth associated protein-43 and microtubule-associated protein 2 mRNA expression in the rat hippocampus. AB - Chronic restraint stress of rats for three weeks produces an atrophy of apical dendrites in the CA3 region of the hippocampus. This alteration is blocked by the novel antidepressant, tianeptine. In order to investigate the underlying mechanism of these phenomena, we evaluated the effect of chronic restraint and tianeptine on mRNA expression of neurotrophic factors such as brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), and basic fibroblast growth factor (bFGF). Chronic restraint and tianeptine treatment did not change the expression of these neurotrophins in the rat hippocampus. We also evaluated the effects of stress and tianeptine on GAP-43 and MAP2, both of which are known to be related to the development of neurons. Chronic restraint resulted in a small decrease in GAP-43 mRNA expression in the CA3 region of the hippocampus, which was not prevented by the concomitant administration of tianeptine. MAP2 mRNA expression was not changed by either chronic stress or tianeptine treatment. We conclude that these neurotrophins, GAP-43 and MAP2 are not likely to be directly related to the chronic stress-induced dendritic atrophy or the prevention of the atrophy by tianeptine. PMID- 9729260 TI - Micronuclei in lymphocyte subsets in relation to immune proteins and allergic disease. AB - This is an investigation of 54 boys and 23 girls with a median age of 19 years (range 18-22 years). The study group contained 12 boys and five girls with asthma and 23 boys and seven girls with allergic rhinitis. Sensitivity to pollen and furred animals were reported by 22 boys and eight girls and by 17 boys and six girls, respectively. The levels of serum immune proteins (IgA, IgE and IgG with subclasses, and IgM) were determined by immunological techniques. As a biomarker of chromosomal damages, the lymphocyte micronuclei was used. We analyzed the frequencies of micronuclei in 3000 B-lymphocytes and in equal numbers of T4- and T8-lymphocytes. The lymphocytes were separated by magnetic attraction in T4 (CD4), T8 (CD8) and B (CD19) fractions using Dynabeads(R). The most interesting finding of this investigation was that the three markers of atopic disease, asthma, hypersensitivity to pollen and IgE levels, associated significantly with increased frequencies of micronuclei in B-lymphocytes. There was also a relation between IgA and the frequency of micronuclei in B-cells. In an epidemiological study of 7000 individuals with allergic diseases, we have found an over-risk for lymphomas in the group with positive skin prick test. Hypothetically, we think that there may be a link between our present finding of an increased mutagenic activity and the lymphoma over-risk among individuals with allergic disease since most lymphomas stem from B-lymphocytes. PMID- 9729261 TI - Effects of repeated nicotine pre-treatment on mesoprefrontal dopaminergic and behavioral responses to acute footshock stress. AB - The effects of acute and repeated nicotine administration on the stress response of rat mesoprefrontal dopaminergic pathways were examined. Rats were given daily injections of nicotine (0.15 or 0.60 mg/kg, s.c., freebase) or saline for 4 days, then challenged with either nicotine or saline. A regimen of inescapable electrical footshocks or no footshocks was then administered. Thirty minutes after final injection, rats were sacrificed, brains removed and dopamine (DA) and its metabolite dihydroxy-O-phenylacetic acid (DOPAC) were extracted from medial prefrontal cortex (mPFC), nucleus accumbens septi (NAS) and dorsolateral striatum and quantified by high performance liquid chromatography with electrochemical detection. Acute administration of low dose nicotine (0.15 mg/kg) produced an increase in DA utilization (increased DOPAC/DA ratio) in mPFC and NAS, but not striatum. High dose nicotine (0.60 mg/kg) produced activation in NAS, but not mPFC or striatum. Repeated low dose nicotine pre-treatment produced tolerance to the effects of nicotine challenge in the mPFC, and reduced its effects in NAS. Footshock stress preferentially increased DA utilization in mPFC and associated footshock stress-induced immobility responses, and these were reduced by low, but not high, dose repeated nicotine pre-treatment. Further, a single dose of the nicotinic acetylcholine receptor (nAChR) antagonist mecamylamine (MCA) 30 min prior to nicotine challenge dose-dependently blocked the reduction of mesoprefrontal DA stress responsivity and immobility responses produced by repeated nicotine pre-treatment. These results indicate that: (1) there are dose dependent differential effects of acute and repeated nicotine pre-exposure on regional DA utilization; (2) low, but not high, dose repeated nicotine reduces both the mesoprefrontal DA and behavioral effects of acute footshock stress; and (3) these effects of repeated nicotine may depend on mecamylamine-sensitive nAChR stimulation. These results may have relevance to acute stress and nicotine dependence, particularly in schizophrenic disorders, which have high prevalence rates of co-morbid nicotine dependence, stress-induced symptom exacerbation and prefrontal cortical dysfunction. PMID- 9729262 TI - Hyperglycemia-exaggerated ischemic brain damage following 30 min of middle cerebral artery occlusion is not due to capillary obstruction. AB - Transient focal ischemia of brief duration (15-30 min) gives rise to brain damage. In normoglycemic animals this damage usually consists of selective neuronal necrosis (SNN), and is largely confined to the lateral caudoputamen. In hyperglycemic subjects damage occurs more rapidly, involves also neocortical areas, and is often of the pan-necrotic type ('infarction'). Since experiments on forebrain ischemia of 30 min duration suggest that microcirculatory compromise develops during recirculation, we studied whether focal ischemia of the same duration, followed by reperfusion for 1, 2 or 4 h, leads to microcirculatory dysfunction. To test this possibility, we fixed the tissue by perfusion and counted the number of formed elements (leukocytes, macrophages and erythrocytes) in capillaries and postcapillary venules. Furthermore, capillary patency was evaluated following in vivo injection of Evan's blue. Histopathological examination of tissue fixed by perfusion after 1, 2 and 4 h of recirculation showed an increasing density of SNN in the caudoputamen of normoglycemic animals. Hyperglycemic, but not normoglycemic, animals showed pan-necrotic lesions ('infarction') after 4 h of recirculation. As a result, the total volume of tissue damage (SNN plus infarction) was larger in hyper- than in normoglycemic animals at 2 and 4 h of recirculation. In addition, hyperglycemic animals showed involvement of neocortex which increased with the time of reperfusion. In the ischemic hemisphere, between 5 and 10% of counted capillaries contained formed elements. However, since hyperglycemic animals contained an equal (or smaller) amount of cells the results did not suggest that capillary 'plugging' could explain the aggravated damage. Moreover, both normo- and hyperglycemic animals showed close to 100% capillary patency. The results thus fail to support the notion that the aggravation of focal ischemic damage by hyperglycemia is due to obstruction of microvessel by swelling or leukocyte adherence. PMID- 9729263 TI - Detection of oxidative mutagenesis by isoniazid and other hydrazine derivatives in Escherichia coli WP2 tester strain IC203, deficient in OxyR: strong protective effects of rat liver S9. AB - Strain IC203, deficient in the OxyR function, was sensitive to both cytotoxic and mutagenic effects of isoniazid (INH) whereas its parent, WP2 uvrA/pKM101, was resistant to these effects. Four other hydrazine compounds, hydrazine hydrate (HZH), phenylhydrazine (PHZ), hydralazine (HLZ) and nialamide (NLD), were mutagenic in WP2 uvrA/pKM101. Increases in mutagenicity were observed in IC203 for HZH and PHZ but not for HLZ and NLD. Growth inhibition zones by HZH, PHZ and NLD were larger in IC203 than in WP2 uvrA/pKM101. The enhancements in the effects of INH, HZH and PHZ in IC203 with respect to its oxyR+ parent are considered to be caused by the production of reactive oxygen species. This is consistent with its inhibition in IC203 by S9 from liver of uninduced rats, probably through the action of catalase. Mutagenicities of INH, PHZ and HLZ were low in strains IC204, a derivative of WP2 uvrA carrying a deletion of the umuDC genes, and IC206, a derivative of IC204 deficient in the MutY glycosylase. In these strains, HZH and NLD induced a high level of revertants which carry suppressor mutations resulting exclusively from G:C-A:T transitions, thus suggesting a direct reaction of the two hydrazines with cytosine. PMID- 9729264 TI - WRYamide, a NPY-based tripeptide that antagonizes feeding in rats. AB - Modifications of (D-Trp32) neuropeptide Y (NPY) led to the development of potential peptide-based lower molecular weight (500-800 Da) NPY feeding antagonists. One compound, WRYamide (N-Ac-Trp-Arg-Tyr-NH2), blocked NPY-induced feeding for 1 to 4 h when injected intrahypothalamically (i.h.t.) at 1 to 40 microgram. Schedule-induced feeding was also antagonized for up to 24 h by 20 microgram of WRYamide, i.h.t. Injection of 2.5 mg/kg (1 mg/rat) of WRYamide, i.v., also reduced significantly schedule-induced feeding for 4 h. A conditioned taste aversion could not be classically conditioned to saccharin using WRYamide as the unconditioned stimulus. These results may lead to the development of systemically active anti-obesity drugs. PMID- 9729265 TI - Links between chromatin structure, DNA repair and chromosome fragility. AB - This paper is a brief overview of the studies we have recently conducted to unravel how chromatin structure and DNA repair modulate the fragility of diverse chromosomes and chromosomal regions. We have employed a combination of molecular cytogenetic techniques, including interphase and metaphase multicolour FISH, reverse FISH with CpG-rich probes or repaired DNA fractions, and several combinations of FISH and immunocytogenetics with antibodies against acetylated histones. The targets of our investigation were human constitutive and facultative heterochromatin, chromosomes with high and low gene density and human and hamster fragile sites. The role of DNA repair was investigated by using DNA repair deficient mutants and DNA repair inhibitors. We found that intragenomic heterogeneity in DNA repair and chromatin structure may explain a substantial part of the differential fragility of diverse chromosomes and chromosomal regions. PMID- 9729267 TI - p53, mutations, and apoptosis in genistein-exposed human lymphoblastoid cells. AB - The phytoestrogen, genistein, is a naturally occurring isoflavone found in soy products. On a biochemical basis, genistein is a competitive inhibitor of tyrosine kinases and the DNA synthesis-related enzyme, topoisomerase-II (topo II). Exposure of mammalian cells to genistein results in DNA damage that is similar to that induced by the topo-II inhibitor and chromosomal mutagen, m-amsa. In order to determine the potential genotoxicity of genistein, human lymphoblastoid cells which differ in the functional status of the tumor suppressor gene, p53, were exposed to genistein and the induction of micronuclei quantified by microscopic analysis. In addition, the mutant fraction at the thymidine kinase (tk) locus (both the normal-growth and slow-growth phenotypes) was determined by resistance to trifluorothymidine (TFT) and at the hypoxanthine phosphoribosyl transferase (hprt) locus by resistance to 6-thioguanine (6-TG). Flow cytometric analysis of the percentage of viable, apoptotic and degenerating cells was utilized to determine the rate and kinetics of cell death after genistein exposure. The detection of micronuclei in both cell lines indicated that genistein-induced damage had occurred in both AHH-1 tk+/- and L3. Linear regression analysis detected a significant increase in the number of 6-TG resistant clones in both AHH-1 tk+/- (p53+/-) and L3 (p53+/+). A comparison of slopes revealed no difference between the lines. In contrast, a significant, concentration-dependent increase in the number of TFT-resistant clones with the slow-growth phenotype was detected in AHH-1 tk+/- (mutant p53), but not in L3 (wild-type p53). Cell death occurred primarily by apoptosis in both cell lines; however, a concentration-dependent decrease in the percentage of viable cells was detected immediately after exposure in L3, but not until 32 h after exposure in AHH-1 tk+/-. A comparison of the slopes of the concentration-response curves for the percentage of viable cells revealed no difference between the cell lines in the effect of genistein on cell viability. Our results may be interpreted that genistein is a chromosomal mutagen and that p53 functional status affects the recovery of chromosomal mutants, possibly by signalling cells into the apoptosis pathways. PMID- 9729266 TI - Potentiation of D2-dopamine receptor-mediated suppression of zif 268 by non competitive NMDA receptor antagonists in reserpinized rats. AB - Striatopallidal output neurons, which coexpress D2-dopamine receptors and NMDA receptors, are logically a potential site of interaction between corticostriatal glutamatergic input and dopaminergic systems. Recent hypotheses about the etiology of schizophrenia have implicated both excitatory amino acid and dopamine systems. The present study was designed to examine, in vivo, the interaction between D2-dopamine receptors and NMDA receptors in the regulation of the expression of the early immediate genes (IEGs), zif 268 and jun B, in striatopallidal neurons. We tested whether coadministration of NMDA antagonists interacted with the actions of the D2 agonist, quinpirole, on IEG expression following dopamine depletion with reserpine. When rats were pretreated with the non-competitive NMDA receptor antagonists, MK 801 (1 mg/kg) or PCP (20 mg/kg), together with quinpirole, the quinpirole reversal of reserpine induction of zif 268 mRNA was potentiated in all regions examined. MK 801 alone had no significant effect on reserpine induction of zif 268 mRNA. Pretreatment with the competitive NMDA receptor antagonist, CPP (5 mg/kg), did not significantly alter the dose response of zif 268 mRNA expression to quinpirole in any region. There was no significant effect of MK 801 on jun B mRNA expression, either on the response to quinpirole or when administered alone with reserpine. Our findings provide evidence of an interaction between the NMDA receptor channel system and the D2 dopamine system on a molecular level in striatopallidal neurons carrying output from the basal ganglia. PMID- 9729268 TI - Nerve growth factor stimulates diacylglycerol de novo synthesis and phosphatidylinositol hydrolysis in pheochromocytoma cells. AB - Induction of neurite outgrowth by treating pheochromocytoma cells (PC12 cells) with nerve growth factor (NGF) is associated with major increases in cellular levels of diacylglycerol (DAG), an essential and probably limiting precursor in phosphatidylcholine (PC) and phosphatidylethanolamine (PE) syntheses. To identify the sources of this DAG we examined the effects of NGF treatment on the conversion of [3H]oleic acid (OA) or [3H]glycerol to [3H]glycerolipids, and the turnover of these products in PC12 cells. In kinetic studies on [3H]OA incorporation, most of the radioactivity in the cells initially was free [3H]OA; then it appeared predominantly as [3H]DAG and, eventually, as large amounts of [3H]phospholipids (PLs). In NGF pre-treated cells, the increases in the levels of [3H]DAG (which were most prominent) and PLs were similar to those in unlabeled DAG and PLs. These effects of NGF could be partially blocked by an inhibitor (triacsin C) of long chain acyl-CoA synthetase. NGF pre-treatment also significantly enhanced the incorporation of [3H]glycerol into lipids, a pathway for de novo synthesis of glycerolipids. In studies on the degradation of [3H]OA labeled lipids, the disappearance of [3H]OA-labeled neutral lipids exhibited an initial rapid phase and a subsequent stable phase. NGF treatment transiently promoted the hydrolysis of [3H]PI to [3H]DAG. These data suggest that the increases in DAG levels observed in PC12 cells exposed to NGF derive mainly from de novo synthesis and, to a lesser and transient extent, from the hydrolysis of [3H]PI. PMID- 9729269 TI - Dose-response curves for X-ray induced interchanges and interarm intrachanges in human lymphocytes using arm-specific probes for chromosome 1. AB - Frequencies of chromosomal aberrations were estimated in X-irradiated (0; 0.5; 1; 2; 3 and 4 Gy) human lymphocytes using arm-specific probes for chromosome 1 in combination with a pancentromeric probe in a triple colour FISH procedure. This allowed the construction of dose-response curves for both interchanges as well as interarm intrachanges. We found that 11.7-18.4% of aberrant chromosomes contained interarm intrachanges. Assuming a free interaction of randomly induced exchange breakpoints throughout the whole genome, interarm intrachanges occur about 8.7 times more frequently than expected. The close proximity of the two arms of a chromosome in the interphase nucleus appears to promote the formation of interarm intrachanges. Convincing evidence was obtained, by comparing the frequency of colour junctions that occur between 1p-1q and 1p-3q (the arms 1q and 3q are of similar size). With this approach, we observed 8.8 times more colour junctions between 1p-1q than between 1p-3q, illustrating the importance of proximity in the formation of chromosomal exchange aberrations. At doses higher than 2 Gy, a saturation of simple reciprocal aberrations, both for interchanges as well as for interarm intrachanges was observed. Furthermore, we found that symmetrical and asymmetrical simple interchanges as well as interarm intrachanges occur with equal frequencies in human chromosome 1. PMID- 9729270 TI - Immunohistochemical and in situ analysis of amyloid precursor-like protein-1 and amyloid precursor-like protein-2 expression in Alzheimer disease and aged control brains. AB - Amyloid precursor protein (APP) is a ubiquitously expressed membrane spanning glycoprotein which is endoproteolytically processed to Abeta, a 39-43 amino acid peptide that is the main component of senile plaques in Alzheimer Disease (AD). APP is a member of a highly conserved gene family, including Amyloid Precursor Like Proteins (APLPs) APLP1 and APLP2. We now characterize APLP1 and APLP2 mRNA and protein expression in AD and aged control brains. Using in situ hybridization in hippocampal tissue from control and AD brain, we show that APLP1 and APLP2 mRNA are expressed primarily in the granule cells of the dentate gyrus, in areas CA1-CA3, and subiculum. Immunohistochemistry reveals staining for both APLP1 and APLP2 in neurons and blood vessels in AD and control cases. In addition, in AD brain, large dystrophic neurites in a subset of senile plaques are conspicuously labeled with APLP1 and APLP2 antibodies. The aged control brains have significantly fewer immunoreactive plaques and dystrophic neurites. The regional, cellular, and subcellular distribution of APLP1 and APLP2 overlap with each other and with APP. These observations support the hypothesis that the members of this family of proteins may perform similar functions. PMID- 9729271 TI - Chromosomal aberrations and alkaline comet assay in families with habitual abortion. AB - Within the pathology of human reproduction, genetic abnormalities play an important role in spontaneous abortions. This paper describes the morphologic, karyotypic features of a consecutive series of singleton spontaneous abortions collected as part of this study and also reports the application of the alkaline comet assay to assess levels of DNA damage in 31 couples comprised of 13 control couples and a patient group of 18 couples with a history of more than one fetal loss. For the cytogenetic analyses, the conventional lymphocyte culture method was applied to all subjects. In this analysis, two women with habitual abortion were determined to carry balanced chromosomal translocation. The alkaline comet assay (single cell gel electrophoresis technique) was applied also to lymphocytes. The comparison of the results of alkaline comet assay in patient and control individuals showed a significant difference in the number of damaged cells. The cells were evaluated according to their grades of damage as: normal (undamaged-no migration), limited migration, (at low damage level) and extensive migration (comet imaged cells-with increasing numbers of breaks, DNA pieces migrate freely into the tail forming a comet image). The frequency of limited migrated and extensive migrated cells in the women in the patient group were higher than in the women in the control group (p<0.001). However, all above parameters were equal for husbands in both the control and patient group (p>0.05). PMID- 9729272 TI - Temporal and spatial changes of quinolinic acid immunoreactivity in the gerbil hippocampus following transient cerebral ischemia. AB - Quinolinic acid (QUIN) is an endogenous neurotoxin which originates from the kynurenine pathway of tryptophan metabolism. An increase of brain QUIN level occurs in several degenerative and inflammatory disorders, but the cellular source of QUIN is still a matter of controversy. In the present study, the gerbil model of transient global ischemia was used to investigate the time course and the cellular localization of QUIN immunoreactivity. Neurodegeneration was evident in the subiculum and in the CA1 area of the hippocampus 4, 7 and 14 days after ischemia. QUIN positive cells, with microglia-like morphology, appeared in the subiculum and in the CA1, 4 days after ischemia. At 7 days post-ischemia they extended to the whole CA1, disappearing at 14 days. Neither neurodegeneration nor QUIN positive cells could be detected in ischemic gerbils sacrificed at 1 and 2 days after ischemia and in sham-operated animals. These findings suggest that microglia-like cells infiltrating the degenerating areas of the hippocampus represent the major source of QUIN following transient ischemia in the gerbil. Thus, in situ production of QUIN in vulnerable brain regions may contribute to the pathophysiological mechanisms of delayed brain injury. PMID- 9729273 TI - Effect of subcutaneous administration of calcium channel blockers on nerve injury induced hyperalgesia. AB - Recent studies suggest that calcium contributes to peripheral neural mechanisms of hyperalgesia associated with nerve damage. In this animal behavioural study, we examined further the contribution of calcium in neuropathic pain by testing whether subcutaneous administration of either a calcium chelating agent or voltage-dependent calcium channel blockers attenuate nerve injury-induced hyperalgesia to mechanical stimulation. Studies were carried out in animals with partially ligated sciatic nerves, an established animal model of neuropathic pain. The nociceptive flexion reflex was quantified using an Ugo Basile Analgesymeter. Partial nerve injury induced a significant decrease in mechanical threshold compared to the sham operated controls. Daily subcutaneous injections of the calcium chelating agent, Quin 2 (20 microgram/2.5 microliter), significantly attenuated the nerve injury-induced hyperalgesia. Similarly, SNX 111, a N-type channel blocker, also significantly attenuated the nerve injury induced hyperalgesia. SNX-230, a P and/or Q-type channel blocker, and nifedipine, a L-type channel blocker, had no effect on the hyperalgesia to mechanical stimulation. In control experiments, SNX-111 had no effect on mechanical thresholds when administered subcutaneously in either the hindpaw of normal animals or the back of the neck in nerve injury animals. This study shows that neuropathic pain involves a local calcium-dependent mechanism in the receptive field of intact neurons of an injured nerve, since it can be alleviated by subcutaneous injections of either a calcium chelating agent or SNX-111, a N-type calcium channel blocker. These agents may be effective, peripherally acting therapeutic agents for neuropathic pain. PMID- 9729275 TI - Protective effects of cortistatin (CST-14) against kainate-induced neurotoxicity in rat brain. AB - Cortistatin (CST-14) is a recently discovered endogenous peptide which shares similarity to somatostatin and binds to somatostatin receptors. In this study, we show that CST-14 exhibits anticonvulsive and neuroprotective effects in rats. Injection of rats with kainic acid (KA; 10 mg/kg; i.p.) generated a strong seizure activity which was attenuated by the i.c.v. application of 1 and 10 nmol CST-14 when given 10 min before KA. Moreover, 3 days after KA injection, a marked loss of neurons in cortex and hippocampus of rats was observed which was inhibited by pretreatment with CST-14. An immunohistochemical analysis using specific antibodies revealed that KA reduced immunoactive sst2A and sst3 somatostatin receptors in the hippocampus-an effect which was largely prevented by pretreatment with CST-14. Superfusion of hippocampal slices with CST-14 also reduced the stimulated release of 3H-d-aspartate. We conclude that CST-14 exerts neuroprotective effects by binding to somatostatin receptors which in turn leads to a reduced release of excitotoxic neurotransmitters. PMID- 9729274 TI - Regulation of G proteins and adenylyl cyclase in brain regions of caffeine tolerant and -dependent mice. AB - Regulation of post-receptor signaling provides a mechanism of adaptation to chronic psychotropic drug treatment. In this study, the regulation of guanine nucleotide binding proteins (G proteins) and G protein-stimulated adenylyl cyclase activity was examined in brain regions of caffeine-tolerant and dependent mice. Chronic caffeine doses were administered via mini-osmotic pumps over 7 days at 0, 42, 85 and 125 mg kg-1 day-1. These chronic caffeine doses were linearly correlated with plasma caffeine concentrations. In behavioral studies, the stimulant effects of acute caffeine on motor activity were significantly diminished in a dose-dependent manner after chronic caffeine, suggesting the development of tolerance. Abrupt discontinuation of chronic caffeine treatment (at 85 and 125 mg kg-1 day-1) produced a dose-dependent and reversible reduction in motor activity 24 h later, suggestive of a caffeine withdrawal syndrome. Utilizing quantitative immunoblotting methods, we found that hippocampal Gialpha1,2 and Gialpha3 subunits were significantly reduced by 20.2% and 11.1%, respectively, in caffeine tolerant/dependent mice (caffeine 125 mg kg-1 day-1 vs. vehicle controls). Decreases in inhibitory G protein subunit concentrations in hippocampus were accompanied by a significant increase (by 21%) in hippocampal G protein function, as measured by guanine nucleotide-stimulated adenylyl cyclase activity, in caffeine-treated mice. This same caffeine treatment also produced significant decreases in cortical Gsalpha subunits of 14.0%. Since short-term caffeine treatment has been shown to reduce adenylyl cyclase activity, chronic caffeine treatment could produce adaptive increases in G protein-stimulated adenylyl cyclase to oppose this effect via G protein regulation. PMID- 9729276 TI - Kinetics of the formation of chromosome aberrations in X-irradiated human lymphocytes, using PCC and FISH. AB - In order to study the initial frequencies and define kinetics of the formation of chromosomal exchanges in X-irradiated human lymphocytes, the premature chromosome condensation (PCC) technique was employed in combination with fluorescence in situ hybridization (FISH) with a composite probe for human chromosome 8 and a pan centromeric probe for the whole genome. Human lymphocytes were X-irradiated (0.5, 1, 2, 3, 4 and 6 Gy), fused with mitotic Chinese hamster ovary (CHO) cells immediately or 1, 3, 6, 12 and 18 h after irradiation. Immediately after irradiation chromosomal breaks, dicentrics and translocations showed a linear dose-response. Unrejoined chromosome breaks were the most frequent types of aberrations (about 85%) observed. About 15% of total aberrations were chromosome exchanges of 65% of these were translocations and 35% were dicentrics. The chromosomal exchanges initially observed were mostly incomplete, with no complex exchanges at doses of 1 and 2 Gy, at higher doses (3-6 Gy) complex exchanges were observed and their frequencies increased with increasing post incubation time. Following different recovery times, repair kinetics of breaks for different doses of irradiation was studied. The shapes of the curves obtained for breaks as well as chromosome exchanges were linear-quadratic. The linear yield component, alpha, is formed entirely in the fast process that can be manifested in the early plateau, while component beta developed slowly in the subsequent hours. The kinetics of breaks rejoining was exponential, almost 50% of breaks rejoined after 1 h and at 18 h about 20% of breaks remained. At low doses of 1 and 2 Gy most of the exchanges were formed immediately and at higher doses, the frequency of exchanges increased with kinetics similar to that observed for the rejoining of breaks. However, the kinetics was different for different doses of irradiation. The frequency of dicentrics increased at doses above 2 Gy following 3 h recovery time, but for the translocations effect was pronounced even at 1 h recovery time. The frequency of incomplete exchanges (i.e., terminal translocations) decreased with post irradiation time and at 18 h was 30-40% less than the frequency obtained immediately after irradiation. The increase in the total translocations as a function of time between irradiation and fusion was due to a rapid increase in complete exchanges (i.e., reciprocal translocations). The frequency of ring chromosomes immediately after irradiation, also increased linearly, however, it was 3-5 times lower than dicentrics and remained almost constant in number for different doses and at different post-irradiation times. PMID- 9729277 TI - Ionizing radiation and genetic risks. VIII. The concept of mutation component and its use in risk estimation for multifactorial diseases. AB - Multifactorial diseases, which include the common congenital abnormalities (incidence: 6%) and chronic diseases with onset predominantly in adults (population prevalence: 65%), contribute substantially to human morbidity and mortality. Their transmission patterns do not conform to Mendelian expectations. The model most frequently used to explain their inheritance and to estimate risks to relatives is a Multifactorial Threshold Model (MTM) of disease liability. The MTM assumes that: (i) the disease is due to the joint action of a large number of genetic and environmental factors, each of which contributing a small amount of liability, (ii) the distribution of liability in the population is Gaussian and (iii) individuals whose liability exceeds a certain threshold value are affected by the disease. For most of these diseases, the number of genes involved or the environmental factors are not fully known. In the context of radiation exposures of the population, the question of the extent to which induced mutations will cause an increase in the frequencies of these diseases has remained unanswered. In this paper, we address this problem by using a modified version of MTM which incorporates mutation and selection as two additional parameters. The model assumes a finite number of gene loci and threshold of liability (hence, the designation, Finite-Locus Threshold Model or FLTM). The FLTM permits one to examine the relationship between broad-sense heritability of disease liability and mutation component (MC), the responsiveness of the disease to a change in mutation rate. Through the use of a computer program (in which mutation rate, selection, threshold, recombination rate and environmental variance are input parameters and MC and heritability of liability are output estimates), we studied the MC-heritability relationship for (i) a permanent increase in mutation rate (e.g., when the population sustains radiation exposure in every generation) and (ii) a one-time increase in mutation rate. Our investigation shows that, for a permanent increase in mutation rate of 15%, MC in the first few generations is of the order of 1-2%. This conclusion holds over a broad range of heritability values above about 30%. At equilibrium, however, MC reaches 100%. For a one-time increase in mutation rate, MC reaches its maximum value (of 1-2%) in the first generation, followed by a decline to zero in subsequent generations. These conclusions hold for so many combinations of parameter values (i.e., threshold, selection coefficient, number of loci, environmental variance, spontaneous mutation rate, increases in mutation rate, levels of 'interaction' between genes and recombination rates) that it can be considered to be relatively robust. We also investigated the biological validity of the FLTM in terms of the minimum number of loci, their mutation rates and selection coefficients needed to explain the incidence of multifactorial diseases using the theory of genetic loads. We argue that for common multifactorial diseases, selection coefficients are small in present-day human populations. Consequently, with mutation rates of the order known for Mendelian genes, the FLTM with a few loci and weak selection provides a good approximation for studying the responsiveness of multifactorial diseases to radiation exposures. PMID- 9729278 TI - Alterations in neuropeptide Y and Y1 receptor mRNA expression in brains from an animal model of depression: region specific adaptation after fluoxetine treatment. AB - To investigate the possible link between neuropeptide Y (NPY) and depression, we analyzed NPY and its receptors in different limbic-related regions in the Flinder sensitive line (FSL), a genetic animal model of depression. In situ hybridization histochemistry was used to measure mRNA expression levels of NPY and NPY receptors, Y1 and Y2, in the FSL as compared to the control Flinder resistant Line rats (FRL). In the FSL rats, NPY mRNA expression levels were significantly decreased in the nucleus accumbens and CA regions, but increased in the arcuate nucleus and anterior cingulate cortex. Y1 receptor mRNA expression was decreased in different cortical regions (retrosplenial, anterior cingulate, and occipital) and in the hippocampal dentate gyrus. Y2 mRNA expression levels did not differ between FSL and FRL animals. The effect of the antidepressant drug fluoxetine (a serotonin reuptake inhibitor) in the two rat strains was also studied. There was an increase of the NPY mRNA hybridization signal in the arcuate nucleus of both strains following the antidepressant treatment (10 micromol/kg; daily for 14 days). However, in other brain regions, fluoxetine administration caused a differential effect on the induction of NPY-related genes in the two rat strains: in the CA region and dentate gyrus NPY mRNA expression was increased in the FSL, but decreased in the FRL. In contrast, Y1 mRNA levels tended to be decreased by fluoxetine in the nucleus accumbens of the FSL rats, but increased in the FRL. These findings suggest an involvement of the Y1, but not the Y2, receptor subtype in depressive disorder. Overall, the results appear to sustain the importance of the FSL rats as an animal model of depression in view of the impairment of NPY genes and the ability of fluoxetine treatment to normalize NPY-related gene expression selectively in this strain. PMID- 9729279 TI - Bcl-2 accelerates the neuronal differentiation: new evidence approaching to the biofunction of bcl-2 in the neuronal system. AB - The proto-oncogene product Bcl-2 is unique in that it inhibits apoptosis rather than promoting cell proliferation. In the present study, we encountered a new possible role of Bcl-2 in the neuronal differentiation. Rat pheochromocytoma PC12 cells have been known as the model of neuronal differentiation by the stimulation of NGF. Bcl-2 transfected PC12 (MB2) cells showed the accelerated neuronal differentiation, as compared with control PC12 (V4) cells. In addition, chemotherapeutic agents Taxol which has been known as neurotoxic compound, induced the acute neuronal cell atrophy and suppressed neuronal differentiation. This neuronal cell atrophy and suppression of neuronal differentiation were not due to apoptotic cell death. Interestingly, Bcl-2 rescued PC12 cells from both neuronal cell atrophy and suppression of neuronal differentiation. Taxol suppressed polymerization between neurofilament light and heavy (NF-L and NF-H), and MB2 cell extract rescued it. We, therefore, suggest the acceleration of polymerization between NF-L and NF-H as the new possible role of Bcl-2. PMID- 9729280 TI - Re-epithelialization of the rabbit cornea is regulated by opioid growth factor. AB - An endogenous opioid peptide, [Met5]-enkephalin, termed opioid growth factor (OGF) is a tonically active, autocrine-produced inhibitory molecule related to developing, neoplastic, renewing and healing tissues. The present investigation was designed to examine the role of OGF on corneal epithelial wound closure in the rabbit under in vitro and in vivo conditions. A 10-mm diameter epithelial defect was made in the center of the rabbit cornea, and the size of the defect, number of specimens with complete re-epithelialization, and rate of wound closure were evaluated using topical fluorescein and morphometric analysis. In organ culture, the influence of a complete opioid receptor blockade by naltrexone (NTX) showed an acceleration in re-epithelialization compared to controls. The action of excessive agonist (OGF) application revealed that exposure of wounded epithelium to OGF delayed wound closure under in vitro conditions, and did so in a receptor-mediated fashion. The modulatory capability of opioids on wound healing in vivo was explored by examining the effects of opioid peptide-receptor disruption using topical application of NTX, and enhanced healing of the abraded rabbit cornea was noted. The presence and location of OGF and the zeta (zeta) receptor in the normal and injured rabbit corneal epithelium were ascertained by immunocytochemistry, and both OGF and the zeta receptor were detected in basal and suprabasal epithelial cells. These results show that an opioid peptide, OGF, plays a direct role in the repair of injury to the corneal epithelium in the rabbit and acts as a receptor-mediated and constitutively expressed inhibitory molecule. PMID- 9729281 TI - Adenosine stimulates stellation of cultured rat cortical astrocytes. AB - We investigated the effect of adenosine on astrocyte morphology by using cell cultures prepared from the cerebral cortices of neonatal rats. Cultured rat cortical astrocytes exhibited flattened, polygonal morphology in the absence of stimulation, but differentiated into process-bearing stellate cells in response to adenosine (1-1000 microM). Adenosine-induced astrocyte stellation was abolished by treatment with microtubule inhibitors, colchicine and paclitaxel, indicating the involvement of cytoskeletal elements. The effect of adenosine was mimicked by other adenosine receptor agonists, and blocked by adenosine receptor antagonists and guanosine 5'-O-(2-thiodiphosphate), indicating that the effect of adenosine is mediated by G protein-coupled adenosine receptors. Although adenosine receptors are known to be linked to adenylate cyclase or phospholipase C, adenosine did not change intracellular cyclic AMP level nor intracellular Ca2+ concentration in astrocytes. Alternatively, adenosine-induced stellation was abolished by tyrosine phosphatase inhibitors, orthovanadate and phenylarsine oxide, suggesting that adenosine causes astrocyte stellation through tyrosine dephosphorylation. Adenosine may function as a factor regulating astrocyte differentiation. PMID- 9729282 TI - Expression and regulation of the human dopamine transporter in a neuronal cell line. AB - Human cocaine users exhibit increased striatal [3H]WIN35428 binding to the dopamine transporter (DAT). However, the nature of the changes induced in the DAT are complex and may not result from a simple increase in number of DAT molecules. To better understand the regulation of DAT inhibitor binding sites and their relationship to the overall process of dopamine uptake, a neuronal model system expressing the human DAT has been developed. Initial experiments were attempted with native dopaminergic neurons so as to allow examination of DAT interactions with vesicular release and storage mechanisms. Dissociated fetal rat mesencephalic neurons, of various ages and mixtures with target cells, were grown to confluence. However, [3H]WIN35428 binding was of low affinity at all levels of maturity. Following this, a simpler model was assessed, using DAT cDNA transfected into neuroblastoma-derived Neuro2A cells. Initially, no specific and little non-specific [3H]WIN35428 or [3H]paroxetine binding was found in non transfected cells. After transfection with the human DAT inserted in the pcDNA vector, both DAT binding and dopamine uptake were significantly and stably present. Treatment with (-)cocaine, 10-6 M for 24 h, increased DAT binding and uptake, which did not occur in parallel COS-7 experiments. Other experiments with Neuro2A cells also found that dopamine uptake was down-regulated by treatment with a PKC activator. These results suggest that the transfected Neuro2A neurons should be useful for ongoing experiments examining the regulation of the DAT by assorted treatments. PMID- 9729283 TI - Relative involvement of chromosome #21 in radiation induced exchange aberrations in lymphocytes of Down syndrome patients. AB - It is not yet resolved as to what type of DNA double strand break repair operates in G0 lymphocytes. We have employed Down syndrome (DS) lymphocytes with three copies of chromosome #21 to answer the question whether the presence of three copies reduces the frequency of exchange aberrations involving this chromosome in comparison to normal cells with two copies of #21. Peripheral blood lymphocytes from three DS patients and two normal individuals were X-irradiated with 1 and 3 Gy. The frequencies of unstable aberrations were found to be higher in DS lymphocytes than normal lymphocytes after 3 Gy of X-rays. FISH studies employing chromosome specific DNA libraries for chromosomes #21 and #22 indicated that the frequencies of exchange aberrations per chromosome are similar in both disomic and trisomic condition. This indicates that the presence of an extra copy of chromosome #21 does not alter the yield, suggesting that homologous recombination does not play a major role in the repair of DNA strand breaks in human G0 lymphocytes. PMID- 9729284 TI - Gender differences in kappa-opioid modulation of cocaine-induced behavior and NMDA-evoked dopamine release. AB - It has been reported that kappa-opioids produce greater analgesia in women than in men. Sex differences are also apparent in drug-induced behaviors. Repeated administration of cocaine (25 mg/kg) produced a greater locomotor and sensitization response in C57BL/6By female mice. It was examined whether the increased sensitization in females to repeated cocaine administration was related to differences in kappa-opioid responses. The effects of the kappa agonist U62066 (spiradoline mesylate) on cocaine-induced locomotor stimulation in vivo and NMDA mediated dopamine release in vitro were measured. In male, but not female mice, U62066 (1 mg/kg) given 30 min before cocaine potentiated the locomotor stimulation of an acute cocaine administration. U-62066 did not affect the development of locomotor sensitization with repeated cocaine administration (25 mg/kg s.c., once daily for 3 days), and a further enhanced response was not seen on days 2 and 3. It was then examined whether dopamine release, measured in vitro, plays a role in sex dependent differences in kappa-opioid- or NMDA modulated dopaminergic function. In tissue perfusion studies, the in vitro NMDA (25 microM)-evoked release of labelled dopamine from striatum was lower in females (fractional release = 5.4 +/- 0.4 and 4.0 +/- 0.4 in male and female mouse striatum). U62066 (1 microM) and ibogaine (1 microM), an indole alkaloid claimed to be useful in the treatment of drug addiction that acts in part at the kappa-opioid receptor, both reduced the NMDA (25 microM)-evoked release of dopamine. Inhibition of the release was significantly greater in tissue from male mice. Prior in vivo cocaine administration did not alter the NMDA-evoked dopamine release. Our studies indicate that kappa-opioid and NMDA receptor activity show differences between female and male mice that may account for differences in cocaine-induced behaviors, but do not exclude the role of other hetereoceptors modulating dopamine release. PMID- 9729285 TI - Characterization of postischemic behavioral deficits in gerbils with and without hypothermic neuroprotection. AB - Five minutes of global ischemia in gerbil results in delayed hippocampal CA1 neuronal degeneration, which is accompanied by working memory impairments and hyperactivity in novel environments. In this study, postischemic activity was characterized in familiar and in novel environments to determine whether hyperactivity was due to impaired spatial habituation or another form of motor hyperactivity. This study also determined whether 6-h delayed hypothermia, which reduces CA1 neuronal injury, would attenuate functional impairments. Gerbils were subjected to 5 min of normothermic ischemia or sham operation 2 days following implantation of brain temperature probes. One of two ischemic groups was cooled (>48 h) starting at 6-h postischemia. Locomotor activity in a familiar cage was measured for 6 days while activity in three novel environments was intermittently measured on days 4, 5 and 6. Open field behavior and working memory in a T-maze were also assessed. Untreated ischemia caused marked hyperactivity in the familiar cage on day 1, which reverted to near-normal by day 2. Nonetheless, these gerbils showed hyperactivity during novel environment sessions on days 4-6. This maze behavior, which predicted hippocampal CA1 injury, was not due to different habituation rates nor baseline hyperactivity. Conversely, open field sessions on day 8 revealed ischemic habituation rate deficits. Ischemia also impaired working memory in the T-maze. Delayed hypothermia, which reduced neuronal loss in the CA1 sector to 12% from 81%, reduced all functional impairments. Ischemic gerbils quickly developed spontaneous locomotion hyperactivity that returned to near-normal after 1 day. This motor hyperactivity did not explain the elevated activity found with delayed testing in novel environments. Regardless, only the open field test on day 8 revealed a habituation-like deficit. PMID- 9729286 TI - Influence of intense sound exposure on glutathione synthesis in the cochlea. AB - Previous studies have shown that depletion of endogenous glutathione (GSH) potentiates noise-induced hearing loss (NIHL), whereas replenishment of GSH attenuates NIHL (Yamasoba et al., Brain Res. 784 (1998) 82-90). Since these findings indicate an important role of GSH in protection from NIHL, we assessed the influence of intense sound exposure (broadband noise, 105 dB SPL, 5 h) on GSH and cysteine levels in the guinea pig cochlea using high performance liquid chromatography. GSH levels were significantly increased in the lateral wall 2 and 4 h post-exposure and returned to normal 6 h post-exposure. GSH levels in the sensory epithelium and modiolus did not show significant changes following noise. Cysteine levels were unchanged in any of the cochlear segments. For the cochlea as a whole, intense sound exposure did not significantly change GSH or cysteine levels throughout the 6-h measurement period post-exposure. These results indicate that GSH synthesis is markedly upregulated selectively in the lateral wall by noise exposure, presumably in response to the robust consumption of GSH, as it is utilized in scavenging reactive oxygen species. PMID- 9729287 TI - Chronic variable stress induces supersensitivity of central alpha2-adrenoceptors which modulate the jaw-opening reflex in the rat. AB - In a previous study, we found that the sensitivity of central postsynaptic alpha2 adrenoceptors which modulate, in an inhibitory way, the activity of the jaw opening reflex (JOR) is reduced after chronic repeated stress (tail pinch) in the rat. The aim of this study was to assess the effects of exposure to a chronic variable stress regime on these adrenoceptors. To do this, the digastric electromyographic responses elicited by orofacial electrical stimulation after the intravenous administration of cumulative doses (x3.3) of the alpha2 adrenoceptor agonist, clonidine (0.1-10000 microgram/kg), were recorded. As expected, in unmanipulated control rats, clonidine inhibited the reflex, in a dose-dependent manner, until abolition (ED50 = 17.3 +/- 2.2 microgram/kg). Single tail pinch did not significantly alter the ability of clonidine to abolish the reflex. However, chronic variable stress led to an enhancement of the inhibitory effect of clonidine on the amplitude of JOR, resulting in a shift to the left of the dose-response curve in comparison with that of the control group (ED50 was reduced by 37%, P = 0.032), without affecting either the estimated maximum effect for the agonist or the slope of the inhibitory function. This in vivo result indicates that chronic variable stress leads to an increased sensitivity of central alpha2-adrenoceptors which modulate JOR, in contrast to the desensitization of these adrenoceptors found after repeated exposure to the same stressor. PMID- 9729288 TI - Role of c-fos in hypoxia-induced AP-1 cis-element activity and tyrosine hydroxylase gene expression. AB - Previous studies have demonstrated that hypoxia stimulates expression of the c fos gene in intact animals and isolated cells. The purpose of the present study was to assess the functional significance of c-fos activation during hypoxia. Using antisense c-fos strategy, we tested the hypothesis that c-fos is essential for activation of activator protein-1 transcription factor complex (AP-1) and subsequent stimulation of down stream genes such as tyrosine hydroxylase (TH) gene during hypoxia. Experiments were performed on rat pheochromocytoma 12 (PC12) cells. AP-1 activity was determined by a reporter gene assay using a luciferase expression vector driven by two copies of an AP-1 cis-element (AP-1-Luc). Cells transfected with AP-1-Luc construct were exposed to normoxia (21% O2) or to varying intensities and/or durations of hypoxia. AP-1 activity increased in response to hypoxia. The magnitude of the response depended on the intensity and duration of the hypoxic stimulus. Increases in AP-1 activity could not be elicited in neuroblastoma cells, indicating that hypoxia-induced increase in AP-1 activity is a cell selective phenomenon. Antisense c-fos abolished hypoxia induced AP-1 activation in PC12 cells. Hypoxia increased tyrosine hydroxylase chloramphenicol acetyl transferase activity (TH-CAT), and antisense c-fos and mutations at AP-1 binding sites in TH promoter abolished this effect. These results provide direct evidence that c-fos is essential for functional activation of AP-1 and subsequent activation of delayed response genes such as TH in PC12 cells. PMID- 9729289 TI - Chromatin structure and chromosome aberrations: modeling of damage induced by isotropic and localized irradiation. AB - Various models for the nuclear architecture in interphase cell nuclei have been presented, proposing a territorial or a non-territorial organization of chromosomes. To better understand the correlation between nuclear architecture and the formation of chromosomal aberrations, we applied computer simulations to model the extent of radiation induced chromosome damage under certain geometrical constraints. For this purpose, chromosomes were described by different models, which approximate the chromatin fiber by a polymer chain, folded in different ways. Corresponding to the different condensation levels, a territorial or a non territorial organization of chromosomes was obtained. To determine the relative frequencies of radiation induced damage, the effects of isotropic ionizing radiation and of a focused laser UV-beam were studied. For isotropic ionizing radiation, the calculated translocation frequencies showed no differences between territorial and non-territorial models except for one special case. For localized irradiation, the results of both organizations were clearly different, with respect to the total number of damaged chromosomes per cell. The predictions agreed well with the experimental data available. PMID- 9729290 TI - Segregation of VIPergic-nitric oxidergic and cholinergic-nitric oxidergic innervation in porcine middle cerebral arteries. AB - The distribution of nitric oxide synthase (NOS)-, choline acetyltransferase (ChAT)-, and vasoactive intestinal polypeptide (VIP)-immunoreactivities, and nicotinamide adenine dinucleotide phosphate diaphorase (NADPHd)-reactivities in the sphenopalatine ganglia (SPG), and perivascular nerves in middle cerebral arteries of the pig was investigated by double-staining techniques using combined immunofluorescence and histochemistry methods. In the SPG, almost all ganglionic cells were NOS-immunoreactive (I) and NADPHd-positive, and both NOS immunoreactivities and NADPHd reactivities were completely co-localized. ChAT-I ganglionic cells accounted for 75%, while VIP-I ganglionic cells represented 42% of all ganglionic cells. Almost all VIP immunoreactivities were co-localized with ChAT immunoreactivities, and all ganglionic cells that were VIP-I and/or ChAT-I were NOS-I and NADPHd-reactive. None of the ganglionic cells in the SPG were immunoreactive to calcitonin gene-related peptide (CGRP). CGRP immunoreactivities, however, were found to surround some ganglionic cells. In middle cerebral arteries, all adventitial NOS-I bundles and fine fibers were coincident with NADPHd fibers. Almost all adventitial ChAT-I bundles and thin fibers, and VIP-I mesh-like fibers stained positively for NADPHd, while the mesh like NADPHd fine fibers were not ChAT-I. Simultaneous labeling using antibodies against VIP and ChAT further indicated that VIP-I fibers were closer than ChAT-I fibers to the smooth muscle. In rare occasions, perivascular fibers were found to be stained for both ChAT and VIP, showing that most ChAT-I and VIP-I fibers were not coincident. These results suggest that ChAT and VIP are rarely co-localized in perivascular nerves in middle cerebral arteries, and point out that the neurotransmitter and the modulator that are co-localized within the same nerve cell body may distribute totally independently and differently at the terminal level. The present results also indicate that in cerebral perivascular nerves, the combination of nitric oxide (NO) and acetylcholine (ACh), as well as the combination of NO and VIP, are localized in the same nerve with different axons containing either NO plus ACh, or NO plus VIP. These findings support the hypothesis that ACh and VIP may act as modulators in regulating presynaptic release of NO, and therefore, cerebral neurogenic vasodilation, from their respective perivascular cholinergic-nitric oxidergic and VIPergic-nitric oxidergic nerves. PMID- 9729291 TI - Effects of lesions in the trigeminal oralis and caudalis subnuclei on different orofacial nociceptive responses in the rat. AB - Behavioural responses to two different orofacial noxious stimulations were analysed following lesion of spinal trigeminal subnucleus oralis (Sp5O) in the rat. Lesions were obtained by intranuclear microinjections of quinolinic acid (0.4 microliter of 60 nmol/microliter solution). The control groups received microinjection of saline. Noxious stimulation was a subcutaneous injection of formalin into the upper lip or electrical stimulation of the tooth pulp. The measured behavioural responses were duration of rubbing induced by the formalin injection and thresholds of the jaw-opening reflex (JOR), head rotation (HR) and face rubbing (FR) evoked by the pulp stimulation. In addition, formalin injection was also performed in two groups of rats that had received intranuclear injection of quinolinic acid or saline into rostral subnucleus caudalis (Sp5C). Rubbing duration was not significantly modified by the lesion of Sp5O, whereas a significant decrease occurred after the lesion of rostral Sp5C. After the lesion of Sp5O, an increase in the threshold of JOR was observed whereas the thresholds of HR and FR were not significantly modified. These results suggest that Sp5O is not necessary for the processing and relay of nociceptive inputs triggered by intense stimulations of oral and perioral areas. However further experiments are needed to reconcile these results with the relevant data obtained from cell recording experiments which indicate the existence, in Sp5O, of neuronal activities related to the sensory discriminative aspect of intense nociception. PMID- 9729292 TI - Distribution and levels of insulin-like growth factor I mRNA across the life span in the Brown Norway x Fischer 344 rat brain. AB - Previous studies have reported changes in insulin-like growth factor I (IGF-I) mRNA expression during early postnatal development of the rat brain. Although changes in IGF-I gene expression have been documented in a wide range of central nervous system structures during early development and investigated in the hippocampus during aging, no study has compared changes in IGF-I gene expression in different brain regions across the life span. The present study assessed the distribution of IGF-I gene expression using in situ hybridization in rats aged 2 30 months. Dot blots were used as a quantitative assessment of cortical IGF-I mRNA. Results indicate that both the distribution and levels of brain IGF-I mRNA do not change significantly between 2 and 30 months of age in the rat. However, in spite of relatively constant levels of mRNA, other studies from our laboratory have demonstrated that cortical IGF-I protein levels decrease 36.6% between 11 and 32 months of age, suggesting that IGF-I function is decreased with increasing age. PMID- 9729293 TI - Effect of maternal folate levels on somatic mutation frequency in the developing colon. AB - Folic acid deficiency is associated with an increase in chromosomal aberrations in adult rodents and humans. Somatic mutations have a critical role in carcinogenesis. Since most mutations arise during early development, the effect of maternal folic acid levels on the spontaneous mutant frequency in the developing colon was examined using lacZ transgenic mice. No significant difference in mutant frequencies at both 3 and 8 weeks of age were found between offspring whose mothers were fed low folate and those on high folate diets during pregnancy. Our results suggest that the correlation between folic acid intake and cancer risk may only be effective at extreme folate deficiencies or in combination with other dietary deficiencies or an underlying predisposition. PMID- 9729294 TI - The alpha5 subunit of the murine type A GABA receptor. AB - GABA[A] receptors in the brain convert binding of GABA (gamma-aminobutyric acid) to inhibition by chloride currents. Several important classes of drugs, including benzodiazepines and alcohol, modulate these receptors, which have also been implicated in epilepsy. We describe the alpha5 subunit of GABAA receptors in mice, comparing inbred DBA/2J mice, prone to juvenile audiogenic seizures, with seizure resistant C57BL/6J mice. We find no sequence differences between the strains, although there are several interesting amino acid differences from the rat. We also compare the expression of the alpha5 subunit in whole brains of DBA/2J mice to that in C57BL/6J mice at 21 days, the peak of the former's seizure susceptibility, again finding no significant difference. We further describe the pattern of expression of alpha5 mRNA during mouse brain development, with a peak at 3 days after birth, and among five brain regions in the adult mouse, with the highest levels in the hippocampus. Finally, we present preliminary evidence for rare alternative splicing of this subunit's message, in the N-terminal extracellular domain, to give a form not translatable into a functional protein. PMID- 9729295 TI - Antisense expression of the human pro-melanin-concentrating hormone genes. AB - Expression of transcripts for human pro-melanin concentrating hormone (pMCH) were studied in the hypothalamus, the primary location for pMCH producing cells in the mammalian CNS. Human hypothalamic tissue was extracted for total RNA and the cDNA generated with reverse transcriptase (RT). PCR amplification with primers spanning exons 2 and 3 of the pMCH human-variant genes (pMCHL), yielded an unspliced product, confirming prior work [T.B. Campbell, C.K. McDonald, M. Hagen, The effect of structure in a long target RNA on ribozyme cleavage efficiency, Nucleic Acids Res. 25 (1997) 4985-4993]. In addition, this product was shown to be exclusively antisense, and to be derived from the 5p (pMCHL1), not the 5q (pMCHL2) locus. Thus, there is no evidence that the MCH peptide-precursor molecule is produced in the brain by the human-variant pMCHL loci. In contrast, corresponding RT-PCR for pMCH RNA generated by the locus on 12q, demonstrated the presence of both sense and antisense spliced RNA. Partial sequencing of the spliced product confirmed that production of at least the two C-terminal peptides would occur from the 12q pMCH locus. The significance of the findings for pMCH and pMCHL1 are discussed relative to what is known about the function of endogenous antisense RNA. PMID- 9729296 TI - Opposing actions of the EGF family and opioids: heparin binding-epidermal growth factor (HB-EGF) protects mouse cerebellar neuroblasts against the antiproliferative effect of morphine. AB - Endogenous opioids and opiate drugs of abuse inhibit the proliferation of cerebellar external granular layer (EGL) neuroblasts by mechanisms that are incompletely understood. Opioids do not act alone, rather multiple extracellular factors regulate granule cell neurogenesis and these undoubtedly act in concert with opioids to shape developmental outcome. We examined whether, heparin binding epidermal growth factor-like growth factor (HB-EGF), a recently described member of the epidermal growth factor (EGF) family, might compete with an inhibitory opioid signal. The results confirmed our ongoing studies that morphine inhibited neuroblast proliferation, while HB-EGF enhanced cell replication. HB-EGF not only counteracted the antiproliferative morphine signal, but invariably enhanced DNA synthesis irrespective of morphine treatment. Our findings suggest that regional and temporal differences in the availability of endogenous HB-EGF may serve to limit the response of EGL neuroblasts to opioids, and HB-EGF may be neuroprotective in opiate drug abuse. If similar responses occur in vivo, then the EGF family and the opioid system may represent distinct and contrasting components of an extracellular signaling system serving to coordinate EGL neurogenesis. PMID- 9729297 TI - Cardiovascular responses to microinjections of glutamate into the nucleus tractus solitarii of unanesthetized supracollicular decerebrate rats. AB - In anesthetized rats, microinjections of excitatory amino acids (EAAs) into the nucleus tractus solitarii (nTS), in a region located immediately rostral to the calamus scriptorius (CS), have been generally reported to elicit depressor and bradycardic responses. On the other hand, in conscious freely moving rats, similar microinjections have been reported to elicit pressor and bradycardic responses. These divergent results have been attributed to the effect of anesthetics. A reinvestigation of the effects of EAAs into the nTS in unanesthetized animals became necessary in order to resolve this controversy. The microinjection technique used in freely moving conscious rats suffers from several technical limitations; for example, microinjections cannot be delivered stereotaxically. In order to avoid these limitations, the present experiments were carried out in unanesthetized supracollicular decerebrate rats. A systematic mapping of nTS in these rats, using microinjections of the solutions of EAAs in artificial cerebrospinal (aCSF) fluid, confirmed that depressor and bradycardic responses are elicited from all the sites in the nTS extending from the CS to a level about 1 mm rostral to it. Pressor responses were elicited by microinjections of l-glutamate (l-Glu) only from the chemoreceptor projection site (a region of the commissural subnucleus, 0.1-0.5 mm caudal to the CS, 0-0.5 mm lateral to the midline and 0.4-0.5 mm deep from the medullary surface). The pressor responses elicited from the aforementioned site were accompanied with bradycardia; this response may be due to diffusion of l-Glu to the dorsal motor nucleus of vagus because the bradycardia disappeared when the depth of the microinjection was reduced to 0.3, instead of 0.5 mm, from the dorsal medullary surface. When urethane was administered intravenously in unanesthetized decerebrate rats, the responses to microinjections of l-Glu remained unchanged, i.e., depressor and bradycardic responses were elicited from all the sites in the nTS extending from the CS to a level about 1 mm rostral to it and pressor and tachycardic responses were elicited from the chemoreceptor projection site. These observations indicated that there is no anesthetic-induced qualitative alteration of the cardiovascular responses to microinjections of EAAs into the nTS. PMID- 9729298 TI - Retrospective dose reconstruction of human radiation exposure by FISH/chromosome painting. AB - Measurements of stable chromosomal aberrations by fluorescence in situ hybridisation (FISH)-chromosome painting has the potential to be used for dose reconstruction, years after exposure to ionising radiation. The method is, however, not yet fully standardised and validated. In particular, with respect to the limited lifespan of circulating T-lymphocytes, a level of uncertainty exists on the long-term persistence of stable aberrations. The main principles of the technique will be demonstrated. Based on results from the literature, the reliability and limitations are discussed. PMID- 9729300 TI - Polymorphic imprinting of the serotonin-2A (5-HT2A) receptor gene in human adult brain. AB - We have examined the imprinting status of the serotonin-2A (5-HT2A) receptor gene (HTR2A) in a series of 41 tissue samples from human adult brain. Using a HpaII RFLP polymorphism in the coding region, we obtained evidence of monoallelic expression of HTR2A in 4 out of 18 informative individuals. However, biallelic expression of HTR2A was observed in other individuals (14/18), suggesting that within the human population HTR2A imprinting in brain is polymorphic. PMID- 9729299 TI - Cytogenetic damage in lymphocytes of healthy and thyroid tumor-affected children from the Gomel region (Belarus). AB - During 1994, 19 thyroid tumor-affected children and 17 healthy children from the Gomel region, one of the areas most polluted by the Chernobyl fallout, were analysed for (i) the presence of in their urine and (ii) chromosome aberrations (CA) in circulating lymphocytes. They were compared with 35 healthy children from Pisa, Italy. Tumor-affected children showed significantly (p<0.05) higher levels in their urine as compared to healthy controls from the Gomel region. No radioactivity was found in urine from the Pisa controls. CA frequency was significantly higher in tumor-affected children compared to the Gomel controls, but was not significantly different between Gomel and Pisa controls. However, dicentric chromosomes were found in a significantly (p<0.01) greater proportion in both affected and healthy Gomel children (3.4 and 1.3/1000 cells, respectively) as compared to the Pisa controls (0.4/1000 cells). Multiple regression analysis showed that the proportion of cells with acentric fragments, dicentric and ring chromosomes was significantly correlated (p<0.05) with the amount of excreted in their urine. These findings suggest that children from the Gomel region were still being exposed to radionuclides, which makes it possible to study a dose-effect relationship. PMID- 9729302 TI - Substance P post-synaptically potentiates glutamate-induced currents in dorsal vagal neurons. AB - We examined the post-synaptic actions of glutamate, N-methyl-D-aspartate (NMDA) and substance P on dorsal vagal neurons, using the patch-clamp technique on brainstem slices of young rats. The vagal neurons were identified electrically and histologically. All vagal neurons responded to glutamate and NDMA and about 30% to substance P, with dose-dependent inward currents. The glutamate-induced currents were blocked partially by either CPP (3((R)-2-carboxypiperazin-4-yl) propyl-1-phosphonic acid) or CNQX (6-cyano-7-nitro-quinoxaline-2,3-dione), indicating that these currents resulted from the activation of at least two types of glutamate receptors: NMDA receptors and AMPA/kainate receptors. The NK1 receptor-selective antagonist, RP67580, blocked substance P-induced currents, suggesting that NK1 receptors do coexist with NMDA receptors and AMPA/Kainate receptors. Substance P potentiated the effects of glutamate. This potentiation lasted 10-20 min and was blocked by CPP and by RP67580, but not by CNQX, demonstrating that the increase in glutamate-induced currents resulted from the interaction between NK1 receptors and NMDA channels. These results provided the first evidence that the receptors for substance P and glutamate coexist on dorsal vagal neurons and interact with each other to modulate visceral efferent functions. PMID- 9729301 TI - Expression of calcium channel alpha1A mRNA and protein in the leaner mouse (tgla/tgla) cerebellum. AB - Homozygous leaner mice carry an autosomal recessive mutation in the Ca2+ channel subunit gene, alpha1A, causing them to exhibit severe ataxia, petit-mal-like epilepsy and a myoclonus-like movement disorder. Expression of alpha1A mRNA in cerebella from 20-day-old homozygous leaner mice was compared to control mice, using in situ hybridization histochemistry. Expression of alpha1A protein was examined in cerebella from 20-day-old homozygous leaner and control mice using immunocytochemistry. No differences in either mRNA or protein expression of the alpha1A subunit were observed when homozygous leaner mice were compared to age matched controls. Therefore, functional alterations in P/Q-Type Ca2+ channels containing the alpha1A subunit need to be explored to further understand the relationship of mutations in the alpha1A gene to the pathogenesis of the neurologic disorders occurring in leaner mice. PMID- 9729304 TI - Good statistics practice (GSP) in genetic toxicology. PMID- 9729303 TI - Novel structure of the human GDNF gene. AB - Glial cell line-derived neurotrophic factor (GDNF) is the most potent known survival factor for substantia nigra neurons, which degenerate in Parkinson's disease, for spinal motoneurons, which die in Lou Gehrig's disease (ALS), and for Purkinje neurons, the critical outflow cells of the cerebellum. Moreover, targeted deletion of the GDNF gene results in renal dysgenesis and abnormal development of the enteric nervous system. GDNF mRNA is expressed in a complex temporospatial pattern in the central nervous system and the periphery, consistent with these observations. To begin elucidating mechanisms regulating the pattern of expression of GDNF, we have cloned the human gene, and characterized the promoter. The promoter is highly GC rich, and lacks canonical CCAT-box and TATA-box motifs. It contains more than 12 binding sites for known transcription factors. These cis-elements have the potential to interact with factors regulating constitutive expression (Sp1) and developmental expression (bHLH). Moreover, the promoter contains sites for binding transcription factors which respond to environmental signals, including CREB, AP2, Zif/268, NFkB, and MRE-BP. Combinatorial actions of these transcription factors may account for the extraordinarily complex expression patterns of the GDNF gene. Importantly, we demonstrate that the hGDNF gene utilizes a promoter distinct from that identified in the rodent GDNF gene, a finding with ramifications for Parkinson's disease and ALS research. PMID- 9729305 TI - Conventional radiation-biological dosimetry using frequencies of unstable chromosome aberrations. AB - Frequency of chromosome aberrations detected by conventional cytogenetics is a very useful parameter in biological radiodosimetry. It can be used for estimating absorbed doses in individuals working with radioactive sources and individuals accidentally exposed to radiation. In the first case subjects wear physical dosimeters as a routine safety habit. Our laboratory at the Institute of Radioprotection and Dosimetry (IRD, Brazil) has been using conventional cytogenetic analysis to complement data obtained by physical dosimetry since 1983. Until now, we have investigated more than one hundred cases where individual physical dosimeters detected occupational exposure (above the safety limits allowed). In total, only 34% of these cases were confirmed by conventional cytogenetic dosimetry. We have also used conventional cytogenetic analysis following the radiation accident of Goiania (Brazil) in 1987. Peripheral lymphocytes from 129 exposed or potentially exposed individuals were analyzed for the frequencies of unstable chromosomal aberrations (dicentrics, centric rings and acentrics fragments) to estimate absorbed radiation doses. During the emergency period, doses were estimated to help immediate medical treatment using in vitro calibration curves produced before the accident. Later on, doses were assessed once more using new in vitro calibration curves. A drawback of this technique is that unstable aberrations are lost after exposure. To investigate the mean lifespan of lymphocytes containing dicentric and ring aberrations, we have followed 15 victims of the Goiania accident over all these years. Results suggest that the disappearance of unstable aberrations is dose-dependent. This could explain the variation in the results found among studies in this field. PMID- 9729306 TI - The alpha4 subunit of rat alpha4beta2 nicotinic receptors is phosphorylated in vivo. AB - The intracellular domains of the alpha4 and beta2 neuronal nicotinic subunits between transmembrane segments 3 and 4 contain a number of predicted phosphorylation sites but there is no direct evidence that any of these sites are actually phosphorylated in vivo. We expressed rat alpha4beta2 nicotinic receptors in Xenopus oocytes, labeled them by an overnight incubation in [32P]orthophosphate, and analyzed the immunoprecipitated receptors by autoradiography and Western blotting. Our results show that the oocytes contained three kinds of alpha4 subunits with apparent weights of 69, 79, and 89 kDa. The 89 kDa alpha4 subunit was the most heavily phosphorylated. PMID- 9729307 TI - Spatial learning and memory induce up-regulation of nitric oxide-producing neurons in rat brain. AB - Changes of nitric oxide (NO)-producing neurons in the brain following learning is not yet clear. In present study, nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) histochemistry and neuronal NO synthase (nNOS) immunohistochemistry were used to detect NOS neurons in rat brain. Results demonstrated that expression of NOS neurons in dentate gyrus and frontal cortex was significantly increased after a water-rewarded spatial alternation task when compared with that after sham training. The elevated expression of NOS neurons occurred not only in the earlier memory stage, but also in the later memory stage. In addition, the expression location and cell counts of NOS neurons in dentate gyrus and frontal cortex with NADPH-d staining or nNOS immunoreactivity resembled each other, but the cell counts of NADPH-d positive neurons were a little more than those of nNOS immunoreactive neurons. The involvement of NO in the processes of spatial learning and memory is further suggested. PMID- 9729308 TI - Micronuclei-biological indicator for retrospective dosimetry after exposure to ionizing radiation. AB - Micronuclei can be measured through a conventional method after staining with Giemsa or fluorescence dyes for DNA. However, a technique with cell proliferation control should be preferred. This is done by incubation with cytochalasin B and counting the micronuclei in binucleated cells. Satisfactory dose relationships are observed after irradiation of human lymphocytes in vitro. The RBE for fast neutrons is around three. An automatic analysis is possible by image analysis. The dose range in which significant increases can be observed is 0.3 to 5 Gy X rays. The assay becomes more sensitive when the micronuclei are determined only in B-lymphocytes. Another possibility exists by determination of the number of micronuclei with centromeres. For this purpose the hybridization with pancentromeric DNA probes and fluorescence labelling is of advantage. By this technique a radiation dose of 0.1 Gy X-rays can be detected. It is apparently also possible under these conditions to detect radiation exposures which have taken place decades before the measurements. PMID- 9729309 TI - Regulation of neuronal nitric oxide synthase mRNA in lordosis-relevant neurons of the ventromedial hypothalamus following short-term estrogen treatment. AB - Short-term estrogenic regulation of neuronal nitric oxide synthase (nNOS) mRNA in the ventrolateral subdivision of the ventromedial nucleus (VLVMN), an area central to lordosis, was demonstrated using in situ hybridization. Estrogen treated animals showed a significantly greater signal in the VLVMN, but not the arcuate or supraoptic nuclei, compared to ovariectomized controls. Neuronal NOS may be involved in early actions of estrogen in the VLVMN. PMID- 9729310 TI - The effect of high-dose albumin therapy on local cerebral perfusion after transient focal cerebral ischemia in rats. AB - We have shown that high-concentration albumin therapy is markedly neuroprotective in focal cerebral ischemia. The present study was conducted to ascertain the degree to which hemodynamic alterations are responsible for this therapeutic effect. Normothermic, physiologically regulated male Sprague-Dawley rats received a 2-h period of middle cerebral artery occlusion (MCAo) by insertion of an intraluminal suture coated with poly-L-lysine. Albumin (25% human serum albumin solution) or vehicle (0.9% sodium chloride) was administered intravenously at a dose of 1% of body weight immediately after suture withdrawal following 2-h MCAo. Local cerebral blood flow (LCBF) was measured autoradiographically with 14C iodoantipyrine after 1 h of recirculation. Novel image-processing methods were used to compare average LCBF data sets against previously obtained infarction frequency data on a pixel-by-pixel basis. Albumin therapy reduced mean hematocrit by 42% but produced no other systemic alterations. Pixel-based histopathological analysis revealed large, consistent cortical and subcortical infarcts in saline treated rats with MCAo; albumin therapy reduced mean cortical infarct volume by 85%. Within regions showing albumin-associated neuroprotection, numbers of pixels having LCBF in the upper ischemic-core flow range (0.12-0.24 ml g-1 min-1) were reduced by 8.6-fold by albumin therapy when compared to saline-treated rats; and numbers of pixels with LCBF in the lower penumbral flow range (0.24-0.36 ml g-1 min-1) were reduced by 3. 1-fold in albumin-treated rats (p=0.04 by repeated measures analysis of variance). Analysis of the [albumin-saline] 3-dimensional difference-image data set revealed a circumferential zone of statistically significant albumin-associated LCBF increase within the posterior portion of the ischemic hemisphere, surrounding the core-region of prior ischemia. Thus, high concentration albumin therapy improves local perfusion to regions of critical LCBF reduction. The spatial extent of this LCBF effect, however, appears too small to account fully for the marked neuroprotective efficacy of this therapy. We suggest that other, non-hemodynamic mechanisms may also be contributory. PMID- 9729311 TI - Estrogen-dependent modulation of rat brain ascorbate levels and ischemia-induced ascorbate loss. AB - Brain ascorbate levels in young adult female rat are lower than those in males. Loss of ascorbate during ischemia is also less in females, suggesting lower oxidative stress. After ovariectomy, however, ischemia-induced loss equals that in males. In the present study, we determined ascorbate levels in maturing male and female rat brain to establish when the gender difference in content arises. We further investigated whether 17beta-estradiol and/or progesterone treatment modulate levels and ischemia-induced loss in ovariectomized females and compared these data with those from normal females in proestrus and estrus. Gender differences in brain ascorbate content were absent before puberty and persisted only in cortex in aging rats. Chronic estradiol treatment, whether alone or in combination with progesterone, prevented an ovariectomy-induced ascorbate increase in hippocampus and caused levels in cortex and cerebellum to fall below those of randomly sampled normal females. These same low levels were found during proestrus and estrus. Estradiol replacement after ovariectomy prevented enhanced ischemia-induced ascorbate loss in hippocampus, but not in cortex or cerebellum. Ischemia-induced losses in proestrus and estrus were similar to those in normal controls. Progesterone had little effect in any region. These data indicate that ascorbate content and redox balance in female brain are influenced postpubertally by estrogens in a region-selective manner. PMID- 9729312 TI - Brain fos-like immunoreactivity in chronic decerebrate and neurologically intact rats given 2,5-anhydro-D-mannitol. AB - Injection of the fructose analogue, 2,5-anhydro-d-mannitol (2,5-AM), increases food intake and Fos-like immunoreactivity (Fos-li) in both brainstem and forebrain structures. Because of the interconnections between brainstem and forebrain areas, it has not been possible to determine whether or to what extent induction of Fos-li in a given region reflects brainstem-forebrain interactions. We addressed this issue using chronic decerebrate (CD) rats with complete transections of the neuroaxis at the meso-diencephalic juncture. CD and neurologically intact control rats were injected (i.p.) with saline or 400 mg/kg 2,5-AM and brains were examined for Fos-li. Both intact and CD rats showed increased Fos-li in the nucleus of the solitary tract (NTS) after injection of 2,5-AM as compared with saline. 2, 5-AM treatment increased Fos-li in the external lateral division of parabrachial nucleus (PBNel) in intact but not in CD rats, suggesting that descending projections from the forebrain may play a role in the activation of PBNel neurons after 2,5-AM injection. Decerebration eliminated significant 2,5-AM-induced Fos-li responses in forebrain structures, including the paraventricular nucleus, supraoptic nucleus, bed nucleus of the stria terminalis and central nucleus of the amygdala. The results are consistent with the hypothesis that the activation of forebrain structures after 2,5-AM treatment is due to stimulation by ascending projections from the brainstem. PMID- 9729314 TI - Cancer chemoprevention from the food-borne carcinogen 2-amino-1-methyl-6 phenylimidazo[4,5-b]pyridine. PMID- 9729313 TI - Mechanisms of radiation-induced chromatid breaks. AB - Chromatid breaks are thought to result from DNA double-strand breaks (dsb) but the mechanisms are not yet understood. The early (but still prevailing) 'breakage first' hypothesis fails to explain the large size of chromatid breaks; many of which are estimated to represent the apparent loss of between 15 and 45 Mbp (up to 30% of an average chromatid). The alternative 'exchange' hypothesis of Revell has potential for explaining the large sizes of deletions, but assumes the interaction of two lesions which therefore predicts a quadratic dependence of chromatid breaks on radiation dose. The exchange hypothesis is not tenable for mammalian cells since chromatid breaks are observed to be induced linearly with dose in both human and rodent cells. An alternative 'signal' model of chromatid breaks is outlined whereby a single dsb, occurring within a large looped chromatin domain, is signalled (possibly by molecules such as DNAPK or ATM protein) and triggers the cell to undergo a recombinational exchange, either within a chromatid or between sister chromatids. If incomplete, such recombinational exchanges would appear as chromatid breaks at metaphase. It is suggested that the large looped chromatin domains could be equivalent to one or more likely several replication 'factories' in which the DNA processing enzymes required for exchange formation would be located. PMID- 9729315 TI - Ten nucleotide cis element in the 3'-untranslated region of the GLUT1 glucose transporter mRNA increases gene expression via mRNA stabilization. AB - The GLUT1 glucose transporter gene is regulated at the post-transcriptional level, and a 10 nucleotide (nt) cis-acting element located at nt 2181-2190 of the GLUT1 3'-untranslated region (3'-UTR) increases the transient expression of a luciferase reporter gene. To investigate the role of this mRNA cis-element, stable transfectants expressing luciferase reporter genes were established in rat C6 glioma cells. Insertion of nt 2100-2300 of GLUT1 3'-UTR resulted in a marked increase in the abundance of both reporter gene mRNA and protein compared to the control, in parallel with a 228% increase in the mRNA t1/2 determined with actinomycin D. Deletion of the 10 nt cis-acting element in the GLUT1 3'-UTR reduced the abundance of reporter gene products and the mRNA t1/2 to levels similar to the control clone. Data suggest that the cis-acting element located at nt 2181-2190 of bovine GLUT1 mRNA 3'-UTR is responsible for increased GLUT1 gene expression via enhanced GLUT1 mRNA stabilization. PMID- 9729317 TI - On the usefulness of drug metabolising enzyme modulation for anti-cancer strategies. PMID- 9729316 TI - [letter]. PMID- 9729318 TI - Radiation induced chromosome aberrations: some biophysical considerations. AB - The implications of recent results using FISH chromosome painting and soft X-ray exposures for the mechanisms of chromosome aberration formation are discussed. It is concluded that the evidence in favour of exchange aberrations arising from one radiation induced chromosome break has increased to the point where a 'change in paradigm' from the older breakage-reunion hypothesis needs to be taken seriously into account. A potential role for recombinational repair of DNA double strand breaks, as known in yeast, in the formation of aberrations in mammalian cells is presented and the relationship between DNA repair studies and radiation cytology is emphasized. PMID- 9729319 TI - Prevention of morphine-induced muscarinic (M2) receptor adaptation suppresses the expression of withdrawal symptoms. AB - Treatment of opiate addiction is generally directed at the suppression of withdrawal symptoms through maintenance of the 'addicted' state with methadone. Yet relatively little is known regarding the neural substrates that contribute to, and maintain the prolonged state of withdrawal experienced by addicts. Opiates can profoundly alter the dynamics of brain and peripheral cholinergic systems, and central administration of anticholinergic drugs in dependent rats has been shown to decrease the expression of precipitated withdrawal symptoms. The purpose of this study was to determine whether the adaptive changes to M2 muscarinic receptors in autonomic centers are linked to the expression of withdrawal phenomena. During the peak period of withdrawal, there was a significant increase in both the expression of M2 muscarinic receptors and its corresponding mRNA within the rostral ventrolateral medulla, a primary vasomotor region. That most of these changes in receptor expression were adaptive in nature was suggested by the fact that when the acetylcholinesterase inhibitor DFP was co administered with morphine, both the increased mRNA expression and the appearance of withdrawal symptoms were inhibited. Thus, interference with morphine-induced M2 muscarinic receptor adaptation in critical brain regions was correlated with a reduction in the development of physical dependence. PMID- 9729320 TI - Tissue and cell type specificity of the human neurotrophin-3 promoter region in transgenic mice. AB - To examine the tissue and cell type specificity of the human neurotrophin-3 (NT 3) promoter, we generated transgenic mice bearing the 3.3 kbp upstream region of the human NT-3 gene linked to the bacterial chloramphenicol acetyltransferase (CAT) gene as a reporter. Eight independent founders of transgenic mice were obtained, and four of them transmitted the transgene to their offsprings. Among three lines of four transgenic mice at 6 weeks of age, a high level of production of CAT protein was detected in the spleen and a low level in the brain. By in situ hybridization analysis, CAT gene expression was detected in the hippocampal neurons and in the cerebellar granular neurons of the transgenic mouse brain. These results suggest that the 3.3 kbp 5' flanking region of the human NT-3 gene has adequate promoter activity in vivo and that its expression pattern resembles the endogenous gene. PMID- 9729321 TI - Reaction of sleep-wakefulness cycle to stress is related to differences in hypothalamo-pituitary-adrenal axis reactivity in rat. AB - Acute stress is known to modify sleep-wakefulness cycle, although with considerable interindividual differences. The origin of these individual differences remains unknown. One possibility is an involvement of the hypothalamo pituitary-adrenal axis (HPA), as its reactivity is correlated with an individual's behavioral reactivity to stress, and it is known to influence the sleep-wakefulness cycle. The present study was designed to analyze relationships between natural differences in behavioral reactivity to stress associated with differential HPA reactivity and stress-induced changes in sleep-wakefulness. Adult rats were classified into two sub-groups according to their locomotor reactivity to a mild stress (novel environment): the 'low responders (LR)' and the 'high responders (HR)' animals exhibited different glucocorticoid secretion in response to stress. We show that immobilization stress induced an increase in wakefulness in LR animals and a decrease in wakefulness in HR animals. On the other hand, paradoxical sleep was increased in both LR and HR animals. Moreover, we observed that LR animals slept more than the HR animals, whereas the two groups had similar levels of paradoxical sleep. These results indicate that the response of the sleep-wakefulness cycle to stress is related to the behavioral reactivity to stress, in turn governed by the individual's reactivity of the HPA axis. The involvement of dopaminergic mechanisms is discussed. PMID- 9729322 TI - An ultrastructural study of p75 neurotrophin receptor-immunoreactive fiber terminals in the reticular thalamic nucleus of young rats. AB - The reticular thalamic nucleus (RT) receives cholinergic fibers from both the basal forebrain and the brainstem. Recent studies have shown that the p75 neurotrophin receptor (p75NTR) is synthesized in cholinergic neurons in the basal forebrain but not in those in the brainstem. In this study, to identify cholinergic fibers originating from the basal forebrain, we used a monoclonal antibody against p75NTR (192-IgG) and characterized the ultrastructure of the immunoreactive fiber terminals in the rostral part of the RT in 3-week-old rats. Light microscopy revealed that p75NTR-immunoreactive fine fibers and varicosities were distributed throughout the nucleus. From electron micrographs, three types of labeled terminals were identified. The first type of labeled fiber terminals (63 out of 106) was consistently small, contained densely packed vesicles, and established asymmetrical synaptic contacts with heavy and bushy postsynaptic thickening on distal dendritic profiles; the second type (18 out of 106) established asymmetrical synaptic contacts with very slight postsynaptic thickening; and the third type (25 out of 106) of labeled terminals contained pleomorphic vesicles and established symmetrical synaptic contacts with more proximal dendritic surfaces than the first two types. In addition to the above, labeled dendritic profiles receiving non-labeled asymmetrical and symmetrical synaptic contacts were identified. These findings suggest that the basal forebrain cholinergic system establishes a variety of synaptic connections in the RT and influences cortical activity indirectly via thalamocortical pathways, as well as via direct projections to the cortex. PMID- 9729323 TI - Co-ordinated and cellular specific induction of the components of the IGF/IGFBP axis in the rat brain following hypoxic-ischemic injury. AB - Insulin-like growth factor 1 (IGF-1) is induced after hypoxic-ischemic (HI) brain injury, and therapeutic studies suggest that IGF-1 may restrict delayed neuronal and glial cell loss. We have used a well-characterised rat model of HI injury to extend our understanding of the modes of action of the IGF system after injury. The induction of the IGF system by injury was examined by in situ hybridization, immunohistochemistry, Northern blot analysis, RNase protection assay and reverse transcriptase-polymerase chain reaction (RT-PCR). IGF-1 accumulated in blood vessels of the damaged hemisphere within 5 h after a severe injury. By 3 days, IGF-1 mRNA was expressed by reactive microglia in regions of delayed neuronal death, and immunoreactive IGF-1 was associated with these microglia and reactive astrocytes juxtaposed to surviving neurones surrounding the infarct. Total IGF-1 receptor mRNA was unchanged by the injury. IGFBP-2 mRNA was strongly induced in reactive astrocytes throughout the injured hemisphere, and IGFBP-3 and IGFBP-5 mRNA were moderately induced in reactive microglia and neurones of the injured hippocampus, respectively. IGFBP-6 mRNA was induced in the damaged hemisphere by 3 days and increased protein was seen on the choroid plexus, ependyma and reactive glia. In contrast, insulin II was not induced. These results indicate cell type-specific expression for IGF-1, IGFBP-2,3,5 and 6 after injury. Our findings suggest that the IGF-1 produced by microglia after injury is transferred to perineuronal reactive astrocytes expressing IGFBP-2. Thus, modulation of IGF-1 action by IGFBP-2 might represent a key mechanism that restricts neuronal cell loss following HI brain injury. PMID- 9729324 TI - Critical steps in alkylation-induced aberration formation. AB - The process leading to chromosomal aberrations as a consequence of DNA damage is probably best understood for simple alkylating agents. Various functions have been identified which are involved in the conversion of critical primary lesions to aberrations. O6-methylguanine is considered to be an important critical preclastogenic DNA lesion, which is converted to aberrations in conjunction with faulty mismatch repair. It operates at sites of O6-methylguanine-thymine mispairing, leading to not yet defined secondary lesions. It is proposed that these secondary lesions cause DNA replication inhibition, which triggers recombination (sister-chromatid exchange formation) and is a critical event involved in aberration production in the second post-treatment replication cycle. For N-alkylations, DNA replication inhibition may result from depurinations and base excision repair intermediates causing a block of replication. In consequence, sister-chromatid exchanges as well as chromosomal aberrations are formed in the first post-treatment replication cycle. Data are available to show that DNA replication inhibition and aberration production in the first post treatment replication cycle are interrelated. PMID- 9729325 TI - Developmental changes in phospholipase D activity and mRNA levels in rat brain. AB - Phospholipase D (PLD) activity and PLD1 mRNA levels were determined in rat brain at ages ranging from embryonic day (E) 19 to postnatal day (P) 49. Basal, oleate , and phosphatidylinositol-4, 5-bisphosphate-stimulated PLD activity increased between E19 and P24 by approximately 3-fold and remained unaltered thereafter. A similar developmental pattern of mRNA levels of PLD1 isoform was found by Northern blotting. The development of PLD correlates with synaptogenesis and myelination suggesting that the enzyme might have an important function in these processes. PMID- 9729326 TI - Effects of prenatal gamma-irradiation on the astrocyte proliferation in response to injury in the brain of 6-day-old rat. AB - Pregnant Wistar rats were exposed to a single 1.0 Gy dose of gamma rays on gestational days 13, 15, 17 or 19 (E13, E15, E17 and E19, respectively). A mechanical injury was made in the cerebral hemisphere of their 6 day-old male offsprings. The injured rats were injected with [3H]thymidine on day 1 or 2 after injury and killed 4 h after the injection. Brain sections were immunostained for glial fibrillary acidic protein (GFAP) or S-100beta protein, subjected to autoradiography and examined microscopically to record proliferating astrocytes. The intensity of astrocyte proliferation in response to injury showed a gradual decrease from the level maximal in brains irradiated on E13 to minimal in those irradiated on E19. The changes were regarded as being related to the stage of prenatal development when irradiation of the brain was performed. PMID- 9729327 TI - Involvement of NMDA receptors and voltage-dependent calcium channels on augmentation of long-term potentiation in hippocampal CA1 area of morphine dependent rats. AB - The involvement of NMDA receptors and voltage-dependent calcium channels on augmentation of long-term potentiation (LTP) was investigated at the Schaffer collateral-CA1 pyramidal cell synapses in hippocampal slices of morphine dependent rats, using primed-bursts tetanic stimulation. The amplitude of population spike was measured as an index of increase in postsynaptic excitability. d, l-AP5 and nifedipine were used as NMDA receptor antagonist and voltage-dependent calcium channel blocker, respectively. The amount of LTP of orthodromic population spike amplitude was higher in slices from dependent rats. Perfusion of slices from control or dependent rats with ACSF containing either D,L-AP5 (25 microM) or nifedipine (10 microM) and delivering tetanic stimulation, showed that D,L-AP5 completely blocked LTP of OPS in slices from both control and dependent rats, while nifedipine attenuated the amount of LTP of OPS in dependent slices and had no effect on control ones. The results suggest that the enhanced LTP of OPS in the CA1 area of hippocampal slices from morphine dependent rats is primarily induced by the NMDA receptors activity and the voltage-dependent calcium channels may also be partially involved in the phenomenon. PMID- 9729328 TI - Generalisability of sensory gating during passive movement of the legs. AB - Movement-related gating of somatosensory evoked potentials in the upper limb is restricted mainly to nerve stimulation supplying the moved limb segment. In the lower limb, this principle may not be followed. Tibial nerve (stimulation at the knee) somatosensory evoked potentials (SEPs) and soleus H reflexes exhibit quite similar patterns of modulation during movement. We hypothesised that movement related gating of initial SEPs in the leg would be generalised from ipsilateral to contralateral leg movement and that such sensory gating would not be generalised to modalities with no functional relevance to the movement. Somatosensory, visual, and auditory evoked potentials (SEPs, VEPs, and AEPs) were recorded from scalp electrodes during unilateral passive movement. Short-latency tibial nerve SEPs, representing the first cortical components, and soleus H reflexes in both the moved leg and the stationary leg were attenuated compared to non-movement controls (p<0.05). Neither VEPs nor middle latency AEPs were modulated (p>0.05). We conclude that sensory gating occurs during contralateral movement. This gating is absent in other sensory modalities with no apparent functional relationship to the imposed movement. PMID- 9729329 TI - DNA double-strand breaks, chromosomal rearrangements, and genomic instability. AB - DNA double-strand breaks can lead to chromosomal rearrangements at the first mitosis after exposure to the DNA strand-breaking agent. The evidence suggests a number of different pathways for DNA double-strand break rejoining in mammalian cells, but it is unclear what factors determine the fate of the induced break and whether or not it will lead to chromosomal rearrangement. If a cell does survive and proliferate after DNA cleavage, delayed chromosomal instability can be observed in the clonal descendants of the exposed cell. Most, but not all DNA double-strand breaking agents are effective at inducing this delayed chromosomal instability. In this paper, we review the evidence for the role of the DNA double strand break in directly induced and delayed chromosomal rearrangements. PMID- 9729330 TI - Vsx-2, a gene encoding a paired-type homeodomain, is expressed in the retina, hindbrain, and spinal cord during goldfish embryogenesis. AB - Vsx-2 encodes a paired-type homeodomain and is the goldfish ortholog of the murine Chx10 gene. During development, Vsx-2 is expressed at high levels in goldfish and zebrafish retina. In addition to the retina, in situ hybridization studies using whole mount and cryosection embryos demonstrate that Vsx-2 is also expressed in subsets of neurons in the hindbrain and in the spinal cord. Expression begins approximately at the metencephalon-myelencephalon border and continues in a restricted lateral zone along the rostral-caudal axis of the spinal cord. These observations suggest a potential requirement for Vsx-2 in the specification and/or the maintenance of neurons in specific CNS regions during embryogenesis. Also discussed are other transcription factors which may act combinatorially with Vsx-2 to regulate neuronal differentiation. PMID- 9729331 TI - Pregnenolone sulfate exacerbates NMDA-induced death of hippocampal neurons. AB - Excessive stimulation of the N-methyl-d-aspartate (NMDA)-type glutamate receptor has been implicated in the neuronal death resulting from focal hypoxia-ischemia. Certain neurosteroids, steroids synthesized de novo in the central nervous system (CNS), have been shown to modulate the action of neurotransmitters at their cellular receptors. Pregnenolone sulfate (PS) is an abundant neurosteroid that enhances the current evoked by NMDA. Using the Ca2+-sensitive fluorescent dye, Fluo-3, AM, and a trypan blue exclusion assay, we evaluated the ability of PS to modulate NMDA-induced changes in intracellular free calcium concentration ([Ca2+]i) and neuronal death in primary cultures of rat hippocampal neurons. The results demonstrate that PS potentiates NMDA-induced increases in [Ca2+]i by 150%. Further, PS exacerbates the MK-801-sensitive neuronal death produced by acute (PS EC50=37 microM) or chronic NMDA exposure, reducing the EC50 of NMDA from 13 to 4 microM under chronic exposure conditions, whereas pregnenolone is ineffective. Our results show that PS, or related sulfated neurosteroids, may play a role in the onset of excitotoxic neuronal death in vivo. PMID- 9729332 TI - UV-induced alterations in the spatial distribution of the basal transcription factor TFIIH: an early event in nucleotide excision repair. AB - New findings concerning the molecular mechanisms of nucleotide excision repair (NER) are discussed. PMID- 9729333 TI - Influence of serum-free culture conditions on subcellular localization of steroid 5alpha-reductase in rat C6 glioma cells. AB - Rat C6 glioma cells are considered to be well characterized, and therefore commonly used as a model system to investigate the function of glial cells. However, recent study has shown that an alteration in the expression of their phenotypic antigens is observed when the cells are maintained under the serum free conditions, proposing the possibility that various properties of glioma cells can be altered by the growth conditions. To test this possibility, the effects of serum-free culture conditions on the expression of steroid 5alpha reductase (5alpha-R) type 1 isozyme in glioma cells were examined using immunocytochemical technique. Immunoreactivity of 5alpha-R type 1 was confined to the perinuclear region of glioma cells cultured in serum-containing medium, and observed in the cytoplasmic space as well as the perinuclear region of the cells cultured in serum-free medium. In contrast, serum deprivation failed to affect the expression of phenotypic antigens, glial fibrillary acidic protein (GFAP) and 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNPase). Further studies showed that the expression of cytoplasmic 5alpha-R immunoreactivity induced by serum deprivation was reversible, and might be attributed to removal of serum proteins rather than biologically active small molecules from culture medium. This alteration in the expression of 5alpha-R immunoreactivity is therefore considered to reflect the translocation of the enzyme from the perinuclear region to the cell cytoplasm rather than the induction of cytoplasmic enzyme, and suggest that the culture conditions cause an alteration in the subcellular localization of 5alpha-R type 1 isozyme without phenotypic change of the glioma cell. PMID- 9729334 TI - Mechanisms of the origin of chromosomal aberrations. AB - This manuscript gives a selective survey of some aspects of the origin of chromosomal aberrations (CA). The following topics are discussed: heterogeneity regarding the induction of initial DNA lesions and their repair among chromosomes or regions of the same chromosome, ratio between symmetrical and asymmetrical exchanges, influence of DNA repeats, chromatin condensation and cell-cycle checkpoints on the formation of chromosome alterations. PMID- 9729335 TI - Reduced number and altered morphology of microglial cells in colony stimulating factor-1-deficient osteopetrotic op/op mice. AB - The numerical density of microglial cells is reduced by 47% in the corpus callosum, by 37% in the parietal cortex and by 34% in the frontal cortex of mice mutant at the op locus which are totally devoid of colony stimulating factor-1 (CSF-1), the major growth factor for macrophages. Moreover, microglia in the frontal cortex of the op/op mice are smaller and have shorter cytoplasmic processes compared to control mice. Study suggests that CSF-1 plays a role in vivo in the formation and maturation of microglia and has little or no effect on perivascular cells. PMID- 9729336 TI - Methylprednisolone inhibition of TNF-alpha expression and NF-kB activation after spinal cord injury in rats. AB - Post-traumatic inflammatory reaction has been implicated in the secondary injury after SCI. TNF-alpha is a key inflammatory mediator, which plays a pathogenetic role in cell death in inflammatory disorders and traumatic brain injury. TNF alpha exerts its effector actions, at least partially, through the activation of a pro-inflammatory transcription factor, NF-kB, which in turn upregulates such genes as iNOS, cytokines, adhesive molecules, and others. Consistent with a post traumatic inflammatory reaction after SCI, we noted an increase in TNF-alpha expression by Western blotting (4.5-fold increase at 1 day after SCI, P<0.01) and immunohistochemistry in a rat SCI model. Post-traumatic TNF-alpha expression was accompanied by an increase in NF-kB binding activity in nuclear proteins isolated from the injured cord (3.9-fold increase, P<0.01). MP is the only drug proven effective in improving neurological function in patients with acute SCI. The mechanism of action of MP is not fully understood, but is thought to be related to its antioxidant effects. MP is also a potent anti-inflammatory agent, which has been recently shown to inhibit NF-kB binding activity. MP (30 mg/kg, i.v.) given immediately after SCI reduced TNF-alpha expression by 55% (P<0.01) and NF kB binding activity. These findings suggest that post-traumatic inflammatory activity that is mediated by the TNF-alpha-NF-kB cascade can be suppressed by MP. PMID- 9729337 TI - Cholinergic function in the hippocampus of juvenile rats chronically deprived of NGF. AB - Intracellular and extracellular recordings were used to assess the cholinergic function in hippocampal slices from juvenile rats chronically deprived of NGF. NGF was neutralised by implanting into the lateral ventricle of postnatal (P) day 2 rats, alphaD11 hybridoma cells (secreting monoclonal antibodies specific for NGF). Parental myeloma cells (P3U) were used as controls. At P15-P18, slow cholinergic EPSPs could be elicited in cells from both alphaD11- and P3U-treated rats. However, slices from alphaD11-implanted rats exhibited a 50% reduction in acetylcholine release following stimulation of cholinergic fibres. This effect was associated to a significant increase in the sensitivity of pyramidal cells to carbachol, as suggested by the shift to the left of the dose/response curve. This may reflect a compensatory mechanism for the reduced efficacy of cholinergic innervation in NGF-deprived rats. In both alphaD11- and P3U-treated rats, carbachol was able to induce a similar concentration-dependent depression of the field EPSPs, evoked by Schaffer collateral stimulation, suggesting that presynaptic muscarinic receptors were not altered. In rats implanted with alphaD11 cells at P15 and sacrificed at P21-P24, no changes in the sensitivity to carbachol were found. At this developmental stage, no differences in acetylcholine release were observed between P3U- and alphaD11-treated animals. These results provide physiological evidence for a regulatory role of NGF in the cholinergic function of the hippocampus during postnatal development. PMID- 9729338 TI - Formation of interleukin-6 in the brain of the febrile cat: relationship to interleukin-1. AB - Previous investigations have shown that interleukin-6 (IL-6), unlike other cytokines, is produced in larger amounts in the brain of the febrile animal regardless of the route, peripheral vs. central, of pyrogen administration. In addition, depending on the experimental condition IL-6 production may or may not require the prior induction of interleukin-1 (IL-1). The present study was carried out in the conscious cat to assess the importance of brain-derived IL-6 in the pathogenesis of fever and the interaction at that site between this cytokine and IL-1. IL-6 was detected in cerebrospinal fluid (CSF) collected at rest and its levels increased during the fever to intravenous (i.v.) endotoxin. The IL-6 elevation, but not the fever, was reversed by pretreatment with intracerebroventricular (i.c.v.) IL-1 receptor antagonist (hIL-1ra). Conversely, when pyrogens (endotoxin, IL-1) were given i.c.v., i.c.v. hIL-1ra reduced the fever without altering significantly the associated rise in CSF IL-6. We conclude that IL-6 is formed in brain in response to both i.v. and i.c.v. pyrogens; however, its formation, whether requiring the prior induction of IL-1 or not, does not appear to be critical for the development of the fever. Blood-borne IL 6, unlike brain-derived IL-6, may still play a role in fever as a trigger of signal-transducing mechanisms operating across the blood-brain barrier. PMID- 9729339 TI - Calcium-dependent nitric oxide production is involved in arsenite-induced micronuclei. AB - Arsenic, a human carcinogen is known to induce sister-chromatid exchanges, chromosome aberrations and micronuclei (MN), but its mechanisms remain unknown. Recently, independent studies have suggested that intracellular calcium and reactive oxygen species are involved in arsenite-induced MN, and nitric oxide (NO) is involved in arsenite-induced poly(ADP-ribosylation). The aim of this research is to investigate the involvement of these molecules in arsenite-induced MN. The intracellular oxidant level and calcium level were monitored with a flow cytometer by using dichlorofluorescein diacetate and fluo3-AM, respectively. The NO production was estimated from the nitrite in cell culture medium with a spectrophotometer by using diaminonaphthalene. The results show that a 4-h treatment with arsenite above 5 microM, caused a dose-dependent increase of oxidant, NO, as well as intracellular calcium level. The arsenite-increased intracellular oxidant level was inhibited by NO synthase inhibitors, S-methyl-l thiocitrulline and Nomega-nitro-l-arginine methyl ester and calcium chelators, ethylene glycol-bis (beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid, and 2-[(2 bis-[carboxymethyl]-amino-5-methylphenoxy)-methyl]-6-methoxy-8- bis[carboxy methyl]aminoquinoline, but not by catalase inhibitor, 3-aminotriazole. The arsenite-increased NO could also be suppressed by NO synthase inhibitors and calcium chelator. However, the arsenite-increased intracellular calcium level was inhibited by calcium chelators, but not by NO synthase inhibitors. A 4-h treatment with arsenite above 10 microM, also induced MN dose-dependently. The arsenite-increased MN could be reduced by NO synthase inhibitors, calcium chelators, as well as superoxide dismutase and uric acid. These results suggest the involvement of peroxynitrite in arsenite-induced MN. We surmise that the disturbance of NO production may cause cardio/peripheral vascular disorders, and the peroxynitrite-mediated DNA damages may cause genetic instability and, hence, cancers in arsenic-exposed humans. PMID- 9729340 TI - Possible involvement of neuropeptide Y in the seasonal control of hydroxyindole-O methyltransferase activity in the pineal gland of the european hamster (Cricetus cricetus). AB - Hydroxyindole-O-methyltransferase (HIOMT) catalyses the last step of all the 5 methoxyindoles synthesized in the pineal gland. The synthetic activity of this neuroendocrine structure is driven not only by noradrenaline but also by various neuropeptides. Recently we have established (1) that one of these neuropeptides, neuropeptide Y (NPY), stimulates specifically HIOMT activity in rat pinealocytes and (2) that the density of the NPY-immunoreactive (NPY-IR) fibers innervating the pineal gland of the European hamster (Cricetus cricetus) displays seasonal variations with a large increase in the late autumn. These findings have led us to evaluate a possible seasonal control of NPY on the European hamster pineal gland. We thus compared the nycthemeral patterns of pineal HIOMT activity and 5 methoxytryptophol (5-ML) content and of circulating MEL levels in European hamsters when NPYergic innervation is low (end of October) and when it is the highest (mid-December). We report in this study that HIOMT activity is significantly increased by 80% in mid-December compared with end of October. This increase is correlated with the appearance of a nycthemeral rhythm of pineal 5-ML levels (with a fourfold increase occurring in early dawn and decreasing slowly towards the end of the day). These observations suggest that NPY could be an important neurotransmitter involved in the seasonal control of the biochemistry of the European hamster pineal gland via a stimulatory effect on HIOMT activity. PMID- 9729341 TI - A brief survey of aberration origin theories. AB - The salient points of three currently debated theories for chromosomal aberration origins (the Classic Breakage-and-Reunion theory, the Exchange theory, and the Molecular theory) are outlined, and some comments are made on each in the light of recent research. PMID- 9729342 TI - The distribution of melanin-concentrating hormone in the monkey brain (Cebus apella). AB - Melanin-concentrating hormone was identified in the brain of Cebus monkey using immunohistochemical and in situ hybridization. MCH-immunoreactive neurons were found in the lateral hypothalamus and dorsolateral zona incerta. MCH-ir fibers were seen in the medial mammillary nucleus, and in the median eminence, and very few fibers in the globus pallidus. This is the first report describing the MCH-ir cell and fiber distribution in the monkey brain. PMID- 9729343 TI - Localization of PKN mRNA in the rat brain. AB - Distribution of mRNA encoding PKN, a fatty acid and RhoA-activated serine/threonine protein kinase with a catalytic domain highly homologous to that of protein kinase C, was investigated in the rat brain using in situ hybridization histochemistry. PKN mRNA proved to be heterogenously distributed. The highest signals were observed in the cerebellum, in limbic systems such as olfactory bulb, hippocampal formation and limbic cortex, and in regions involved in central autonomic and neuroendocrine functions, such as hypothalamic ventromedial, dorsomedial, lateroanterior and arcuate nuclei, paraventricular hypothalamic nucleus and locus coeruleus. PKN mRNA was also highly expressed in dopaminergic neurons such as the ventral tegmental area and substantia nigra pars compacta, in serotonergic raphe neurons, and in cholinergic neurons such as nucleus diagonal band, nucleus basalis, and lateral dorsal tegmental nucleus. The distribution of PKN mRNA differed from that for PKC isoforms. As the localization of PKN mRNA is heterogeneous, PKN may have a specific role in distinct populations of nerve cells. PMID- 9729344 TI - Regional distribution of subtypes of nicotinic receptors in human brain and effect of aging studied by (+/-)-[3H]epibatidine. AB - Epibatidine, a potent nicotinic agonist, was used to study the regional distribution of nicotinic acetylcholine receptor binding sites in the human brain. Saturation studies performed in the human temporal cortex with (+/-) [3H]epibatidine revealed binding to two binding sites with Kd and Bmax values of 0.018 and 4.2 nM, 12.7 and 15.4 fmol/mg protein, respectively. Competition studies with (+/-)-[3H]epibatidine/unlabelled nicotine or [3H]nicotine/unlabelled (+/-)-epibatidine showed binding to two binding sites in the human temporal cortex (Ki=0.16 and 12.6 nM; 0.007 and 0.3 nM, respectively). Similarly, when unlabelled nicotine was used to displace (+/-)-[3H]epibatidine, two binding sites were also revealed in the thalamus and the cerebellum of human brain (Ki=0.065 and 7.7 nM; 0.07 and 12.5 nM, respectively). The regional binding of (+/-) [3H]epibatidine binding in human brain was somewhat different from that of [3H]nicotine. A proportionally higher binding was observed for (+/-) [3H]epibatidine in the cerebellum and the thalamus compared to [3H]nicotine, probably reflecting different selectivity to nicotinic receptor subtypes. A marked significant age-related decrease in (+/-)-[3H]epibatidine binding was observed in the frontal and the temporal cortices (-79%, -84%, respectively) of human subjects between 56-85 years of age, which was similar to that of [3H]nicotine (-82%, -79%, respectively). The (+/-)-[3H]epibatidine binding in the cerebellum decreased significantly with age (-77%), while [3H]nicotine binding showed no significant age-related changes in this brain region. The findings indicate that a specifically modulate regional nicotinic receptors in human brain. PMID- 9729345 TI - Chronic ethanol administration alters activity in ventral tegmental area neurons after cessation of withdrawal hyperexcitability. AB - The present study investigated the activity of neurons in the mesolimbic dopamine system after the end of the acute phase of the behavioural signs of ethanol withdrawal in mice. This was designed to provide a comparison with earlier behavioural studies, in which greater development of sensitisation to amphetamine and cocaine, but no change in the initial effects of these compounds, or in the behaviour in the absence of drug treatment, was seen when repeated injection of these psychostimulants were given after chronic ethanol consumption. In the present study, single unit recordings were made from dopamine-sensitive neurons in the ventral tegmental area in perfused midbrain slices prepared 24 h after cessation of chronic ethanol consumption. Profound decreases in firing of the ventral tegmental area (VTA) neurons were seen in slices prepared after the ethanol treatment. Firing rates increased after application of N-methyl-dl aspartate, but still remained lower and more variable after the ethanol treatment. Application of dopamine or amphetamine, following stimulation of firing with a low concentration of N-methyl-dl-aspartate, also resulted in lower firing rates in slices from ethanol-treated mice. No changes were seen in release of tritiated dopamine, in response to applied KCl or amphetamine, from slices of striatum or cerebral cortex, prepared 24 h after cessation of the chronic ethanol consumption, compared with control values. The results demonstrate that very substantial decreases in firing rate, and in the number of active cells, occur in VTA neurons at a time when withdrawal hyperexcitability was no longer apparent and overt changes in behaviour were not seen. PMID- 9729346 TI - Neurotransmitter actions on oral pontine tegmental neurons of the rat: an in vitro study. AB - The actions of neurotransmitters involved in the sleep-wakefulness cycle on neurons located in the ventral part of the oral pontine tegmentum were studied in a rat brain-slice preparation. Results show that glutamate and histamine evoke depolarizations and spike firing while serotonin and gamma-aminobutyric acid evoke hyperpolarizations. The excitatory and inhibitory actions of these neurotransmitters increase pontine neuron activity during the conditions occurring during paradoxical sleep. PMID- 9729348 TI - Tectal projections to the parvicellular reticular formation and the upper cervical spinal cord in the rat, with special reference to axon collateral innervation. AB - After Phaseolus vulgaris leucoagglutinin injection into the lateral part of the superior colliculus (SC) in the rat, labeled fibers and axon terminals in the lower brainstem were distributed not only in the medial reticular formation but also in the lateral tegmental field including the parvicellular reticular formation (RFp). More caudally, in the upper cervical spinal cord labeled fibers with bouton-like varicosities were distributed mainly in laminae V, VII and VIII, with relatively sparse distribution in lamina IX. These labeled axons were found bilaterally with a clear-cut contralateral dominance. After combined injections of rhodamine-dextranamine, Fluoro-ruby (FR) into the RFp and Fluoro-gold (FG) into the upper cervical spinal cord on the same side, SC neurons labeled with FR were intermingled with those labeled with FG in the lateral part of the SC contralateral to the injection sites. In the stratum griseum intermediale, some of them were double-labeled with both tracers. Our results suggest that SC neurons innervating both the RFp and the cervical spinal cord may be involved in the coordination of head and mouth movements. PMID- 9729347 TI - Increased activity improves recovery of partially denervated fast rat muscles. AB - Partial denervation of the neonatal rat extensor digitorum longus muscles by removing the L4 spinal nerve and thus 80% of its innervation [A.L. Connold, T.J. Fisher, S. Maudarbocus, G. Vrbova, Response of developing fast muscles to partial denervation, Neuroscience 46 (1992) 981-988; F. Tyc, G. Vrbova, The effect of partial denervation of developing rat fast muscles on their motor unit properties, J. Physiol. 482 (1995) 651-660] results in its permanent weakness. The possibility that the weakness that follows partial denervation is due to the effects of reducing activity of the muscle during a critical stage of development was studied here. Partial denervation was carried out in 3-day-old pups by removing the L4 spinal nerve. To enhance motor activity two days later some animals had injections of l-Dopa twice a day for 8 days. This treatment induced locomotor activity for at least 2 h/day. The muscles from treated and untreated animals were examined 2 months later. There was a significantly smaller reduction of weight and force production in the muscles from l-Dopa treated animals. Both twitch and tetanic force developed by the EDL muscle from the treated group was twice that of the control untreated group. This effect was due mainly to the larger size of the motor units (MUs) in the l-Dopa treated muscles compared to the controls. The mean motor unit force in the untreated group was 69% of that in the control muscle, whereas this value was 127% in the l-Dopa treated animals. Thus it appears that the activity induced by treatment with l-Dopa could to some extent prevent the loss of weight and force output seen after partial denervation of young fast muscles. PMID- 9729350 TI - An automated new technique for scoring the rodent micronucleus assay: computerized image analysis of acridine orange supravitally stained peripheral blood cells. AB - We developed an automated image analysis system to obtain objective data for the rodent peripheral blood micronucleus assay with acridine orange (AO) supravital staining. The system was able to identify micronucleated reticulocytes (MNRETs) and to evaluate inhibition of bone marrow cell proliferation by measuring the reticular area of reticulocytes (RETs). We also developed automated equipment to produce homogeneous acridine orange-coated glass slides. This study was designed to compare automated scoring with manual scoring using 4 model clastogens and 2 mouse strains. The MNRET incidence induced by each clastogen was similar for automated and manual scoring, and there was good correlation (r=0.92) between the methods. In addition, an index of bone marrow toxicity based on the reticular area of RETs was compared to the conventional index (% of polychromatic erythrocytes (PCE) to total erythrocytes; PCE ratio) and was similar. The results indicated that our technique for computer-assisted image analysis for the micronucleus assay with AO supravitally stained peripheral blood RETs was comparable to conventional microscopic scoring, and it was superior in objectivity and statistical power. PMID- 9729349 TI - Inhibitory effect of threo-9,10-dichlorostearic acid on the mutagenic activity of MeIQx, 2-AF and B[a]P in the Ames/Salmonella test. AB - The mutagenic activity of threo-9,10-dichlorostearic acid, one of the chlorinated fatty acids identified in fish lipids, was examined in the Ames/Salmonella test. No mutagenic activity was found on any of the Salmonella typhimurium strains TA 98, TA 100 and TA 102, either with or without S9 activation. On the other hand, dichlorostearic acid showed an inhibitory effect on the mutagenic activity of the indirectly-acting mutagens 2-amino-3, 8-dimethylimidazo[4,5-f]quinoxaline (MeIQx), 2-aminofluorene (2-AF) and benzo[a]pyrene (B[a]P) using strain TA 98 in the presence of S9. However, no inhibition was observed when mixing MeIQx and S9 before the addition of dichlorostearic acid. Furthermore, dichlorostearic acid did not show any inhibitory effect on the mutagenic activity of the directly acting mutagen 4-nitroquinoline-N-oxide (4NQO) using the tester strains TA 98 and TA 100. We, therefore, suggest that dichlorostearic acid interacts with the enzymes of the S9 mix, thereby dose-dependently inhibiting the transformation of MeIQx, 2-AF and B[a]P into their active forms. PMID- 9729351 TI - Thyroid hormones in human tumoral and normal nervous tissues. AB - We have studied T4 and T3 concentrations, DNA and protein concentrations and 5' and 5 deiodinases in samples of brain tumors obtained at surgery from 49 patients, and, in most cases, also from surrounding normal tissue. T4 concentrations in normal cortical tissue (6.19+/-0.45 ng/g) were lower than in white matter, but the difference disappeared when referred to the DNA content (2.26+/-0.27 ng/mg DNA). No other differences were found between cortical and white matter, or among cortical lobes. T4 in normal tissue was higher than previously reported, mostly from autopsy samples, whereas T3 (0.99+/-0.07 ng/g) was similar. 5'D-I activity was negligible as compared to 5'D-II (8.11+/-1.09 fmol/h/mg protein). When expressed in relation to the different DNA contents of normal vs. tumoral tissue, 5'D-II activities were the same for both. 5D activity was highly variable in the tumoral tissue, with negligible activities in meningiomas and pituitary adenomas. When referred to the DNA content, T4 and 5'D II were the same, but T3 concentrations were lower in the tumor (0.24+/-0.03 ng/mg DNA) as compared to normal (0.35+/-0.04 ng/mg DNA) tissue samples. Whether or not this decrease of T3 affects the expression of T3-sensitive processes remains to be studied. PMID- 9729352 TI - Neurosteroid modulation of recombinant ionotropic glutamate receptors. AB - Pregnenolone sulfate (PS) is an abundant neurosteroid that can potentiate or inhibit ligand gated ion channel activity and thereby alter neuronal excitability. Whereas PS is known to inhibit kainate and AMPA responses while potentiating NMDA responses, the dependence of modulation on receptor subunit composition remains to be determined. Toward this end, the effect of PS on recombinant kainate (GluR6), AMPA (GluR1 or GluR3), and NMDA (NR1(100)+NR2A) receptors was characterized electrophysiologically with respect to efficacy and potency of modulation. With Xenopus oocytes expressing GluR1, GluR3 or GluR6 receptors, PS reduces the efficacy of kainate without affecting its potency, indicative of a noncompetitive mechanism of action. Conversely, with oocytes expressing NR1(100)+NR2A subunits, PS enhances the efficacy of NMDA without affecting its potency. Whereas the modulatory efficacy, but not the potency, of PS is increased two-fold by co-injection of NR1(100)+NR2A cRNAs as compared with NR1(100) cRNA alone, there is little or no effect of the NR2A subunit on efficacy or potency of pregnanolone (or epipregnanolone) sulfate as an inhibitor of the NMDA response. This suggests that the NR2A subunit controls the efficacy of neurosteroid enhancement, but not inhibition, which is consistent with our previous finding that potentiating and inhibitory steroids act at distinct sites on the NMDA receptor. This represents a first step towards understanding the role of subunit composition in determining neurosteroid modulation of ionotropic glutamate receptor function. PMID- 9729354 TI - Important variables that influence base-line micronucleus frequency in cytokinesis-blocked lymphocytes-a biomarker for DNA damage in human populations. AB - The cytokinesis-block micronucleus (CBMN) assay has been adopted by numerous laboratories as a means for rapidly assessing base-line chromosome damage (breakage and loss) in human populations. However, the appropriate implementation of this assay requires a thorough understanding of both experimental variables and biological factors that can have impact on micronucleus (MN) frequency. The paper describes, with the help of experimental data the from the author's laboratory as well as other data, the impact of these variables. With regards to experimental variables, the scoring of micronuclei on slides by different technicians has been identified as an important factor; however, the use of different culture media, namely RPMI 1640 and McCoy's medium, did not have a significant effect on base-line frequencies. The paper also describes results showing that the MN index in cytokinesis-blocked cells, measured once every three months over a 12-month period for 53 healthy subjects, remains constant and the data measured on these occasions were significantly and positively correlated (R=0.477 to 0.684, P<0. 0001) with each other thus indicating the reliability and intra-individual variability of the assay over time. Inter-individual variation for males and female subjects has been estimated for each decade of age between 20 and 80 years; the difference between the 25th and 75th percentile of MN frequency varied between 1.4 fold and 2.3 fold and the minimum and maximum values for MN frequency varied by a factor of 4.7 and 12.5 depending on the age group. Age and gender are the most important demographic variables impacting on the MN index with MN frequencies in females being greater than those in males by a factor of 1.2 to 1.6 depending on the age group. For both sexes, MN frequency was significantly and positively correlated with age (R=0.62 in males and R=0.65 in females) and the slope of the regression line in males was 0.314 (P<0.0001) and in females it was 0.517 (P<0.0001). The main dietary factors influencing the MN index in subjects who are not folate deficient are plasma B12 (R=-0.315, P=0.0127) and plasma homocysteine (R=0.415, P=0.0086). In addition, it was proposed that the MN index is likely to be influenced by the propensity of an individual's cells to undergo apoptosis when damaged so that one might expect the MN frequency to be negatively correlated with apoptotic rate although this has yet to be tested. The above indicates the importance of maintaining an international network of scientists working with the CBMN assay to ensure appropriate quality control and for the development of standard experimental and documentation protocols. The human micronucleus (HUMN) project launched in 1997 is briefly described and proposed as the vehicle for these activities. PMID- 9729353 TI - BDNF, and full length and truncated TrkB expression in the hippocampus of the rat following kainic acid excitotoxic damage. Evidence of complex time-dependent and cell-specific responses. AB - Systemic administration of kainic acid (KA) at convulsant doses results in irreversible cell damage and neuron loss in the hilus of the dentate gyrus and in the CA1 area of the hippocampus. This is followed by reactive astrocytosis in these regions, and sprouting of mossy fibers into the molecular layer of the dentate gyrus. Since trophic factors are probably implicated in the cellular responses to the excitotoxic insult, and early induction of BDNF and TrkB mRNAs has been observed following KA injection, the present study examines BDNF, full length and truncated TrkB protein expression in the hippocampus, as revealed by immunohistochemistry, up to 30 days following KA administration to adult rats. Reduction in BDNF and full-length TrkB immunoreactivity preceding neuron loss is observed in the damaged areas. However, transient increase in BDNF immunoreactivity is observed in surviving CA1 neurons and in granule cells of the dentate gyrus. In contrast, full-length TrkB immunoreactivity progressively increases in the molecular layer of the dentate gyrus up to day 30 following KA administration. A second peak in BDNF immunoreactivity is observed in reactive astrocytes, as revealed with double-labeling immunohistochemistry to BDNF and GFAP, in the plexiform layers of CA1 and, to a lesser degree, in the molecular layer of the dentate gyrus. In addition, strong truncated TrkB immunoreactivity is found in reactive astrocytes, as revealed with double-labeling immunohistochemistry to truncated TrkB and GFAP, in the same regions. These results, in concert with previous observations in the same model of hippocampal damage, suggest that BDNF participates in the early response to excitotoxic damage, and that expression of full-length TrkB at strategic sites in the molecular layer of the dentate gyrus has a role in the regenerative response linked to mossy fiber sprouting. Interestingly, delayed expression of BDNF and truncated TrkB in reactive astrocytes may act as negative regulators of neurite growth in devastated regions, such as the CA1 area, which are impoverished of putative postsynaptic sites. PMID- 9729355 TI - Effects of 7-nitroindazole, a neuronal nitric oxide synthase (nNOS) inhibitor, on locomotor activity and contextual fear conditioning in rats. AB - The effect of 7-nitroindazole (7-NI), a selective neuronal nitric oxide synthase (nNOS) inhibitor, on contextual fear conditioning in rats was examined. Systemic administration of 7-NI did not affect the acquisition of contextual fear (measured as freezing), but it did reduce locomotor activity and cause a corresponding increase in the expression of contextual freezing. It is concluded that nNOS activity is not required for either the acquisition or expression of contextual fear conditioning. PMID- 9729356 TI - The expression of tenascin-C along the lamprey olfactory pathway during embryonic development and following axotomy-induced replacement of the olfactory receptor neurons. AB - Extracellular guidance molecules affect the pathway of growing axons by both attractive and repulsive interactions. Tenascin-C, a glycoprotein of the extracellular matrix, is localized along developing axonal pathways where it may function by repulsion, restricting axons within specific boundaries. The lamprey olfactory pathway offers an advantageous model for studying the role of extracellular matrix proteins in axon guidance because the entire pathway is readily seen in horizontal sections and because lesioning the olfactory nerve will induce the system into a new phase of coordinated neurogenesis and axon outgrowth. Although tenascin-C expression was absent during embryonic development, olfactory nerve fascicles contained tenascin-C-immunoreactivity (IR) during the larval stage. During retrograde degeneration, the fascicles lost tenascin-C-IR. Diffuse unfasciculated axonal processes extending from the olfactory epithelium did not express tenascin-C-IR; however, acetylated tubulin and GAP-43-IR was present, indicating axonal outgrowth. When the newly extended axons of olfactory receptor neurons converged to form fascicles, tenascin-C-IR was evident within the fascicular boundaries. The absence of tenascin-C expression when axonal process were short and diffuse, and its return when axons coalesced within fascicles, supports the view that tenascin-C functions as a boundary molecule in the olfactory pathway. PMID- 9729357 TI - ABT-594, a novel cholinergic channel modulator, is efficacious in nerve ligation and diabetic neuropathy models of neuropathic pain. AB - A novel cholinergic channel modulator, ABT-594, was tested in two established and distinct models of neuropathic pain; the Chung model (i.e., tight ligation of L5 and L6 spinal nerves) and a diabetic neuropathy model (i.e., streptozotocin induced diabetes). Tactile allodynia and mechanical hyperalgesia were assessed in the Chung and diabetic neuropathy models, respectively. ABT-594 produced a significant antiallodynic effect following both oral (0.1-1 micromol/kg) and intraperitoneal (i.p.) (0.3 micromol/kg) administration. Equal efficacy was observed following both routes of administration. ABT-594 (0.3 micromol/kg, i.p.) maintained efficacy following repeated dosing (5 days; twice daily) in the Chung model, but the effect of morphine (21 micromol/kg, i.p.) was significantly reduced after repeated dosing. In the diabetic neuropathy model, ABT-594 (0.3 micromol/kg, i.p.) effectively reduced mechanical hyperalgesia. Morphine (21 micromol/kg, i.p.) was not effective in this model. Overall, these results suggest development of ABT-594 may provide a novel pharmacotherapy for the chronic treatment of neuropathic pain. PMID- 9729358 TI - Chromium can reduce the mutagenic effects of benzo[a]pyrene diolepoxide in normal human fibroblasts via an oxidative stress mechanism. AB - The interaction of multiple carcinogens on human cells has not been extensively examined. This study reports the results of experiments in which normal human fibroblasts were exposed to both benzo[a]pyrene diolepoxide (BPDE) and potassium dichromate. The effect of four different treatment protocols on the cloning ability of the cells and the mutant frequency of the HPRT gene was determined. The combined treatment of both carcinogens caused a slightly greater than additive decrease in the cloning ability of the cells when compared to cells treated with the individual carcinogens. The result was the same regardless of the treatment protocol used in the experiment. The results of the mutant frequency experiments, however, varied dramatically with the protocol employed. The mutant frequency in cells which were simultaneously treated with both carcinogens was dramatically reduced from the mutant frequency found when cells were treated with BPDE alone. This antagonistic effect was not present when cells were either pretreated with potassium dichromate prior to BPDE or incubated with potassium dichromate following BPDE treatment. The observed antagonistic effect was the result of oxidative stress produced by chromium since it was completely or nearly completely reversed by the addition of either vitamin E or catalase to the cultures. PMID- 9729359 TI - Immobility and flight associated with antinociception produced by activation of the ventral and lateral/dorsal regions of the rat periaqueductal gray. AB - It has long been known that the periaqueductal gray (PAG) plays an important role in the modulation of nociception. Given that activation of the lateral PAG also produces wild running and tachycardia, it has been suggested that PAG mediated antinociception is part of an integrated defensive reaction. However, an alternative hypothesis is that these effects are merely a secondary response to aversive brain stimulation. If antinociception and flight reactions are caused by aversive brain stimulation, then these effects should always occur together. The objective of the present study was to determine whether antinociception and locomotion could be dissociated by microinjecting morphine and kainic acid into various subdivisions of the caudal PAG. Non-selective activation of lateral and dorsal regions of the PAG by microinjection of kainic acid produced wild running, while injections into the ventrolateral PAG produced immobility. Microinjection of morphine evoked similar locomotor effects, although the onset to effect was slower with morphine (approximately 5 min vs. 1 min for kainic acid), and the antinociceptive efficacy of microinjecting 0.2 microl of morphine was less than with kainic acid injections. In fact, microinjection of morphine evoked locomotor effects in the absence of antinociception on 39% of the tests. Increasing the injection volume to 0.4 microl (dose remained at 5 microg) greatly enhanced the likelihood that antinociception and locomotor effects (e.g. running, freezing, circling) occurred simultaneously (79%). These findings indicate that, although distinct locomotor effects are associated with antinociception from the ventral and more dorsal regions of the PAG, antinociceptive and locomotor effects can occur independently. This finding is consistent with the hypothesis that ventral and dorsal regions of the PAG integrate defensive freezing and flight reactions, respectively. PMID- 9729360 TI - Characterization of blink reflex interneurons by activation of diffuse noxious inhibitory controls in man. AB - The blink reflex consists of an early, pontine R1-component and a late, medullary R2-component. R1 and R2 can be evoked by innocuous stimuli, but only the R2 also by painful heat, suggesting that the R2 is mediated by wide dynamic range neurons (WDR) of the spinal trigeminal nucleus. Remote noxious stimuli suppress the activity in WDR neurons via activation of diffuse noxious inhibitory controls (DNIC), whereas low-threshold mechanoreceptive neurons (LTM) are unaffected. In order to characterize the trigeminal interneurons of R1 and R2 we investigated the modulation of the blink reflex by remote painful heat. The blink reflex was elicited in 11 healthy subjects by innocuous electrical pulses applied to the left supraorbital nerve. The remote, painful heat stimuli were applied by a Peltier type thermode to the left volar forearm. Remote painful heat of 44 to 46 degreesC significantly suppressed the R2 by 15% (p<0.01), while the R1 remained unchanged. These results provide further evidence that the R2 is mediated by medullary WDR neurons and the R1 by pontine LTM neurons. PMID- 9729361 TI - Central infusion of glucagon-like peptide-1-(7-36) amide (GLP-1) receptor antagonist attenuates lithium chloride-induced c-Fos induction in rat brainstem. AB - Central infusion of glucagon-like peptide-1-(7-36) amide (GLP-1) and intraperitoneal (i.p.) injection of lithium chloride (LiCl) produce similar patterns of c-Fos induction in the rat brain. These similarities led us to assess the hypothesis that neuronal activity caused by i.p. injection of LiCl involves activation of central GLP-1 pathways. We therefore determined if third ventricular (i3vt) infusion of a GLP-1 receptor antagonist would block LiCl induced c-Fos expression in the brainstem. Relative to rats pretreated with i3vt infusion of vehicle, pretreatment with the potent GLP-1 receptor antagonist, des His1 Glu9 exendin-4 (10.0 microgram), significantly attenuated LiCl-induced (76 mg/kg; i.p.) c-Fos expression in several brainstem regions, including the area postrema, the nucleus of the solitary tract, and the lateral parabrachial nucleus. While central infusion of des His1 Glu9 exendin-4 also blocked GLP-1 induced (10.0 microgram) anorexia and c-Fos expression, the antagonist produced no independent effects on food intake or c-Fos expression. These results suggest that LiCl-induced c-Fos expression in the rat brainstem is mediated, at least in part, by GLP-1 receptor signaling. PMID- 9729362 TI - Functional characterization and visualization of a GABAA receptor-GFP chimera expressed in Xenopus oocytes. AB - The GABAA receptor is a ligand-gated chloride channel belonging to the superfamily of ligand-gated ion channels of which the nicotinic acetylcholine (nACh) receptor is prototypic. In the central nervous system the GABAA receptor mediates fast neuronal inhibition. To facilitate the study of this receptor, a GABAA receptor-green fluorescent protein (GABAAR-GFP) chimera was constructed by fusing green fluorescent protein (GFP) to the C-terminus region of the GABAA receptor alpha1 subunit. When expressed in Xenopus oocytes, this chimera responded in a manner indistinguishable from the wild-type GABAA receptor with respect to agonist potency, receptor desensitization, allosteric modulation, rectification, and ion selectivity of the channel. The addition of GFP to the GABAA receptor alpha1 subunit did not appear to alter the assembly or efficiency of expression of the GABAA receptor complex. The GABAAR-GFP chimera generated a strong fluorescent signal that was restricted to the animal pole of the oocyte plasma membrane. This signal was readily detectable using either epifluorescence or laser confocal microscopy. To confirm the extracellular location of the GFP portion of the chimera, non-permeabilized oocytes were immunolabeled with an anti GFP antibody. Fluorescence microscopy showed that GFP was located extracellularly since it was accessible to the GFP antibody. These results confirm the predicted extracellular location of the C-terminus of the GABAA receptor alpha1 subunit and also demonstrate that GFP retains its fluorescent property when expressed extracellularly. The usefulness of the GABAAR-GFP chimera in receptor trafficking was investigated using non-hydrolyzable GTP analogues since GTP binding proteins participate in protein transport in oocytes. Microinjections of GTP-gamma-S but not GDP-beta-S reduced both GABA-gated chloride currents and cell surface GFP fluorescence in oocytes expressing the GABAAR-GFP chimera indicating that the chimera undergoes internalization upon stimulation of oocyte GTP-binding proteins. The results of the present study show that the GABAAR-GFP chimera is functionally similar to the wild-type GABAA receptor and can be used to study receptor trafficking in living cells. This is the first demonstration of a ligand gated ion channel-GFP chimera for an ion channel belonging to this superfamily and also is the first example of the fusion of GFP to an extracellular domain of an integral membrane protein. PMID- 9729363 TI - Chromosome aberration assays in genetic toxicology testing in vitro. AB - The chromosome aberration test using cultured mammalian cells is one of the sensitive methods to predict environmental mutagens and/or carcinogens, and is a complementary test to the Salmonella/microsome assay (Ames test). From our recent survey of 951 chemicals which have been tested for their clastogenicity in cultured mammalian cells such as Chinese hamster fibroblasts or human lymphocytes, it was noted that 47% of them are consistently positive either with or without metabolic activation. When the test was performed using the cell line CHL/IU, 39.2% (292/745) were found to be positive. However, 8% (36/447) of such clastogens were positive only at an extremely high concentration of more than 10 mM. About 11% (48/447) of clastogens such as diethylstilbestrol (DES) and methyl AalphaC (Glob-P-1) induced mainly polyploid cells. Most chemicals induced chromatid-type aberrations, some induce only break-type aberrations at relatively high dose levels, but others induce more exchange-type aberrations at relatively low dose levels. Clastogenic activities were compared among different clastogens, using the D20 value, which is the minimum dose (mg/ml) at which aberrations were found in 20% of metaphases. In addition, the translocation (TR) value was calculated from the incidence of cells with exchange-type aberrations. It was suggested that possible carcinogens are included in the group of compounds with relatively low D20 values, but with high TR values. Karyological analysis was performed, using a FISH painting probe prepared from No. 1 chromosome of CHO cells, on the clonal subline isolated after treatment with benzo(a)pyrene. However, no specific changes common to the agent were detected. Laser scanning cytometry (LSC) was also applied to screen for abnormal karyotypes. A translocation between particular chromosomes was reflected by the deletion of a DNA peak. PMID- 9729364 TI - Involvement of mu- and delta-opioid receptors in the ethanol-associated place preference in rats exposed to foot shock stress. AB - The purpose of this study was to establish the ethanol-induced place preference in rats exposed to foot shock stress using the conditioned place preference paradigm. We also investigated the role of the endogenous opioid system in the development of the ethanol-induced place preference. The administration of ethanol (300 mg/kg, i.p.) with foot shock stress, but not without such stress, induced a marked and significant place preference. Naloxone (1 and 3 mg/kg, s.c.), a non-selective opioid receptor antagonist, significantly attenuated the ethanol-induced place preference. Moreover, the selective mu-opioid receptor antagonist beta-funaltrexamine (3 and 10 mg/kg, i.p.) and selective delta-opioid receptor antagonist naltrindole (1 and 3 mg/kg, s.c.), but not the selective kappa-opioid receptor antagonist nor-binaltorphimine (1 and 3 mg/kg, i.p.), significantly attenuated the ethanol-induced place preference. Furthermore, 150 mg/kg ethanol (which tended to produce a place preference, although not significantly) combined with each dose (that did not produce a place preference) of the mu-opioid receptor agonist morphine (0.1 mg/kg, s.c.) or selective delta opioid receptor agonist 2-methyl-4aalpha-(3-hydroxyphenyl)-1,2,3,4,4a,5,12, 12aalpha-octahydroquinolino [2,3,3-g] isoquinoline (TAN-67; 20 mg/kg, s.c.), but not the selective kappa-opioid receptor agonist trans-3, 4-dichloro-N-(2-(1 pyrrolidinyl)cyclohexyl)benzenacetamide methanesulfonate (U50,488H; 1 mg/kg, s.c.), produced a significant place preference. These data indicate that stress may be important for development of the rewarding effect of ethanol, and that mu- and delta-opioid receptors may be involved in the rewarding mechanism of ethanol under stressful conditions. PMID- 9729365 TI - Temporal evolution of neuronal changes in cerebral hypoxia-ischemia in developing rats: a quantitative light microscopic study. AB - Studies in adult animal models of transient cerebral hypoxia-ischemia (HI) and ischemia suggest that morphologic evidence of neuronal death in some regions such as striatum appears early, while in other regions such as cerebral cortex and CA1 region of hippocampus it is delayed for few days and is referred to as delayed neuronal death (DND). Moreover, in some regions such as CA2/CA3 early 'reactive' neuronal changes occur that are potentially reversible. The aim of this study was to determine whether such changes may also occur in the developing brain. To that end, unilateral cerebral HI was produced in postnatal rats of 13, 21, and 30 days (p13, p21, p30) by right common carotid artery ligation and hypoxemia (breathing 8% O2), and their brains were examined at 24 h, 36 h, 72 h, and 96 h of recovery. The results suggest that: (i) DND is present in developing brain, but its regional distribution varies with animals' age. In cerebral cortex, it is more pronounced in p30 rats than in younger animals. In hippocampus, comparison of lesions of similar severity at different age groups shows a more pronounced DND in CA2/CA3 region of p13 rats than in older animals, but no significant differences exist in the degree of DND in CA1 neurons among different age groups. (ii) 'Reactive' neuronal changes characterized by reduction in Nissl staining and acidophilia of neuronal perikaryon with minimal nuclear abnormality are present at 24 h of recovery. These changes in some regions, such as in CA1 and cortex, progress to neuronal death, while in other regions such as in CA2/CA3 are potentially reversible. (iii) Recovery of reactive neurons in CA2/CA3 region is age dependent in that there is significant recovery in the older age groups, but not in p13 rats. The pathogenetic mechanisms of the reactive neuronal changes, the chain of events leading to DND or neuronal recovery, and the influence of age in these processes remain to be elucidated. PMID- 9729366 TI - Comparisons of chemically-induced mutation among four bacterial strains, Salmonella typhimurium TA102 and TA2638, and Escherichia coli WP2/pKM101 and WP2 uvrA/pKM101: collaborative study III and evaluation of the usefulness of these strains. AB - Over the past 5 years, a large collaborative study of chemically-induced mutation has been performed using the four bacterial strains Salmonella typhimurium TA102 and TA2638 and Escherichia coli WP2/pKM101 and WP2 uvrA/pKM101 in order to compare the specific spectrum of response to chemicals and to evaluate the usefulness (sensitivity) of each strain. Following the two collaborative studies to test the chemicals in category 1, chemicals previously judged as positive only in E. coli WP2 strains and derivatives of these chemicals, and category 2, oxidative agents or crosslinking agents, 22 compounds of category 3 consisting of 10 nonmutagenic carcinogens and another 12 chemicals were selected in this study. Twenty participating laboratories tested each compound in the same method as previous reports. In the group of nonmutagenic carcinogens, no chemical induced revertant colonies of any strain tested. In the group of other chemicals, response to the chemicals was similar in TA102 and WP2 uvrA/pKM101. Overall, in the three collaborative studies, a total of 79 compounds were tested. No difference in qualitative response to the four strains was observed for 71% (56/79) of the test chemicals. The combination of strains providing the greatest number of positive responses was WP2 uvrA/pKM101 with TA102; 84% (66/79) of the test chemicals elicited the same qualitative response in these two strains. Therefore, it is suggested that WP2 uvrA/pKM101 and TA102 can be included as a part of the standard tester strains for detection of mutagenic activity of chemicals. PMID- 9729367 TI - Immunohistochemical analysis of transferrin receptor: regional and cellular distribution in the hypotransferrinemic (hpx) mouse brain. AB - The hypotransferrinemic (hpx) mouse mutant produces <1% of the normal circulating level of transferrin (Tf). Heterozygote animals of this strain (hpx/+) have approximately 50% of normal plasma Tf levels. In this study we examine the cellular and regional distribution of Tf receptor (Tf-R) in the brain of wild type, hpx/+ and mutant (hpx/hpx) mice. Also, using slot-blot (immunoblot) analysis, we describe the relative amount of Tf-R in brain microvessels of hpx/+ animals compared with wild type. Tf-R was seen primarily in neurons throughout the brains of wild type, hpx/+ and hpx/hpx animals. Gray matter areas immunoreacted more robustly than white matter areas. Oligodendrocytes and third ventricle tanycytes, both of which we have previously described as iron-positive, did not immunoreact for Tf-R. Tf-R immunohistochemical reaction in wild type, hpx/+ and hpx/hpx brains appeared similar. Immunoblot analysis of isolated cortical microvessels from wild type and hpx/+ animals revealed no upregulation of Tf-R expression in hpx/+ (relative to normal) despite a 50% decrease in circulating Tf levels. These results indicate that Tf-R is primarily expressed by neurons and that half normal levels of Tf (hpx/+) or transferrin supplementation (hpx/hpx) are apparently sufficient for normal expression and distribution of Tf R. Because of the lack of circulating Tf, but unaltered Tf-R expression, hpx mice could serve as a model for delivery of therapeutic agents via the Tf/Tf-R system. PMID- 9729368 TI - Chromosome aberration testing in genetic toxicology-past, present and future. AB - In vitro metaphase tests for chromosomal aberrations (CA) have undergone considerable evolutionary changes over the last 20 yr. Treatment and sampling times have been a particular focus of attention as we have tried to develop protocols that detect weak genotoxins. Different approaches evolved in different parts of the world and led to a need to harmonise. At the same time, we have increasingly challenged the conditions in which clastogens produce positive responses, and several situations have been described in which clastogenic responses would be considered not to be biologically relevant. Now there is a strong case to replace the conventional metaphase analysis test with an in vitro micronucleus test. The time is therefore right to carefully consider whether the type of damage scored in CA tests is relevant for human health. PMID- 9729369 TI - Orphan neurones and amine excess: the functional neuropathology of Parkinsonism and neuropsychiatric disease. AB - The aetiology and treatment of Parkinsonism is currently conceptualised within a dopamine (DA) deficiency-repletion framework. Loss of striatal DA is thought to cause motor impairment of which tremor, bradykinaesia and rigidity are prominent features. Repletion of deficient DA should at least minimise parkinsonian signs and symptoms. In Section 2, based on extensive pre-clinical and clinical findings, the instability of this approach to Parkinsonism is scrutinised as the existing negative findings challenging the DA deficiency hypothesis are reviewed and reinterpreted. In Section 3 it is suggested that Parkinsonism is due to a DA excess far from the striatum in the area of the posterior lateral hypothalamus (PLH) and the substantia nigra (SN). This unique area, around the diencephalon/mesencephalon border (DCMCB), is packed with many ascending and descending fibres which undergo functional transformation during degeneration, collectively labelled 'orphan neurones'. These malformed cells remain functional resulting in pathological release of transmitter and perpetual neurotoxicity. Orphan neurone formation is commonly observed in the PLH of animals and in man exhibiting Parkinsonism. The mechanism by which orphan neurones impair motor function is analogous to that seen in the diseased human heart. From this perspective, to conceptualise orphan neurones at the DCMCB as 'Time bombs in the brain' is neither fanciful nor unrealistic [E.M. Stricker, M.J. Zigmond, Comments on effects of nigro-striatal dopamine lesions, Appetite 5 (1984) 266-267] as the DA excess phenomenon demands a different therapeutic approach for the management of Parkinsonism. In Section 4 the focus is on this novel concept of treatment strategies by concentrating on non-invasive, pharmacological and surgical modification of functional orphan neurones as they affect adjacent systems. The Orphan neurone/DA excess hypothesis permits a more comprehensive and defendable interpretation of the interrelationship between Parkinsonism and schizophrenia and other related disorders. PMID- 9729370 TI - Nurr1 mRNA expression in neonatal and adult rat brain following kainic acid induced seizure activity. AB - Nurr1 is an immediate early gene encoding a member of the steroid-thyroid hormone receptor family. In PC12 cells, Nurr1 is readily induced by membrane depolarization, but not by growth factors. Nurr1 is predominantly expressed in the brain, and is essential to the differentiation of midbrain dopaminergic neurons. However, Nurr1 is also expressed in brain regions unrelated to dopaminergic neurons, e.g., hippocampus and cerebral cortex, and its immediate induction following seizure activity suggests a potential involvement of this transcription factor in modulating gene expression in the nervous system. To investigate the response of Nurr1 to neuronal activation, we analyzed Nurr1 mRNA expression in neonatal and adult rat brain following kainic acid (KA)-induced seizure. In P7 animals, systemic injection of KA increased Nurr1 mRNA levels in a few hilar cells of the dentate gyrus and some pyramidal cells of the CA3 region of the hippocampus. In older animals, Nurr1 induction progressively expanded to all hippocampal regions (P14, P21) and eventually to cortical regions (adult). The increase was rapid and transient in the dentate gyrus, a structure resistant to the neurotoxic effect of KA, and was more prolonged in other regions more susceptible to KA toxicity. Induction of Nurr1 at early postnatal stages and rapid increase in the dentate gyrus following KA-induced seizure, suggest that Nurr1 expression is modulated by neuronal activity. On the other hand, prolonged Nurr1 induction in regions sensitive to KA toxicity indicates a possible involvement of Nurr1 in selective neuronal vulnerability. PMID- 9729371 TI - Sleep-waking discharge patterns of ventrolateral preoptic/anterior hypothalamic neurons in rats. AB - Numerous lesion, stimulation and recording studies in experimental animals demonstrate the importance of neurons within the preoptic/anterior hypothalamic area (POA) in the regulation of sleep induction and sleep maintenance. Recently, a discrete cluster of cells in the ventrolateral POA (vlPOA) of rats was found to exhibit elevated c-fos gene expression during sleep, indicating that these neurons are strongly activated during nonREM and/or REM sleep stages. We examined neuronal discharge during wakefulness and sleep throughout the dorsal to ventral extent of the lateral POA in rats, using chronic microwire technique. We found that neurons with elevated discharge rates during sleep, compared to waking, were localized to the vlPOA. As a group, vlPOA neurons displayed elevated discharge rates during both nonREM and REM sleep. Discharge of vlPOA neurons reflected the depth of sleep, i.e., discharge rates increased significantly from light to deep nonREM sleep. During recovery sleep following 12-14 h of sleep deprivation, vlPOA neurons displayed increased sleep-related discharge, compared to baseline sleep. Neurons in the vlPOA displaying increased neuronal discharge during sleep were located in the same area where neurons exhibit increased c-fos gene expression during sleep. Such neurons are likely components of a rostral hypothalamic mechanism that regulates sleep onset and sleep maintenance. PMID- 9729372 TI - Reversal of abnormalities of myelination by thyroxine therapy in congenital hypothyroidism: localized in vivo proton magnetic resonance spectroscopy (MRS) study. AB - Deficiency of thyroid hormone during central nervous system ontogeny results in a variety of clinical, anatomical and biochemical defects. Delay in thyroxine therapy in newborns with congenital hypothyroidism leads to irreversible brain damage. We have used localized in vivo proton magnetic resonance spectroscopy (MRS) to assess biochemical changes in different regions of brain in three patients with congenital hypothyroidism before and after thyroxine therapy. An abnormal lipid peak which disappeared with thyroxine therapy was observed in cerebellum and frontal lobe in one patient. Statistically significant reduction of NAA/(Cr+PCr) [P<0.009] and elevation of Cho/(Cr+PCr) [P<0.008] ratios in comparison to controls were documented in all three patients which tended to normalise with thyroxine therapy. A variety of biochemical abnormalities relatable to myelin maturation were documented and these were found to be reversible on thyroxine therapy. Reversibility was documented even though thyroxine therapy was initiated at ages beyond which abnormalities in myelinogenesis are considered irreversible. Also, proton MRS revealed biochemical heterogeneity between patients with congenital hypothyroidism. PMID- 9729374 TI - Opioid receptor gene expression in the rat brain during ontogeny, with special reference to the mesostriatal system: an in situ hybridization study. AB - The three main types of opioid receptors micro, delta and kappa are found in the central nervous system and periphery. In situ hybridization study was undertaken to determine the expression of mu, delta, kappa-opioid receptors mRNAs in the brain during pre- and postnatal development, especially in the mesostriatal system. By G13, mu and kappa-opioid receptor mRNA were detectable in the telencephalon; mu-opioid receptor mRNA was found in the striatal neuroepithelium and cortical plate and kappa-opioid receptor mRNA in the corroidal fissure. By G15, kappa-opioid receptor mRNA was detectable in the nucleus accumbens and dorsal striatum, and in the substantia nigra and ventral tegmental area, suggesting an early expression of the corresponding receptor on dopaminergic terminal fibers. For the mu-opioid receptor mRNA in the striatum, patches appeared at G20. Delta-opioid receptor mRNA was first detected at G21, in many areas including the accumbens nucleus and the dorsal striatum. At P8, delta opioid receptor mRNA was detected in large-sized cells of the striatum, possibly cholinergic, suggesting a possible modulation by opioids of the striatal cholinergic neurons. Our results demonstrate the early appearance of mu and kappa opioid receptor mRNA (G13) and the relatively late development of delta-opioid receptor mRNA (G21) in the brain. We also show a distinct pattern of expression for mu, delta and kappa-opioid receptor mRNAs in the mesostriatal system during the development. PMID- 9729373 TI - Unique activation of extracellular striato-pallidal neurotransmitters in rats following acute risperidone. AB - The present study investigated the effects of the putative atypical antipsychotic drug (APD), risperidone, on striatal monoamine and pallidal gamma-aminobutyric acid (GABA) function using dual probe in vivo microdialysis. Risperidone (0.03, 0.3, 3 mg/kg) or vehicle was injected (s.c.) into female, Sprague-Dawley rats fitted with dual microdialysis probes in the striatum and the globus pallidus (GP). In the striatum, risperidone increased extracellular levels of dopamine (DA) and 3,4-dihydroxyphenylacetic acid (DOPAC) at all doses and the serotonin (5 HT) metabolite, 5-hydroxyindoleacetic acid (5-HIAA), at the highest dose. The increase in striatal DA was most pronounced at the lowest dose of risperidone; however, DOPAC showed a dose dependent increase. Risperidone at the medium and high doses significantly reduced extracellular GABA levels in the GP. Simultaneous measurement of limb rigidity during microdialysis showed that risperidone dose-dependently produced significant increases in horizontal bar test catalepsy and fore- and hindlimb paw retraction latencies. The current results suggest novel effects of risperidone on striatal DA release, while the pallidal GABA changes are similar to previous results obtained with the atypical antipsychotic drug, clozapine. Additionally, the behavioral results predict the clinical expression of extrapyramidal motor side effects at high doses. Overall, these results support an atypical profile of risperidone when compared with typical APDs, yet one with unique neurochemical and behavioral properties. PMID- 9729375 TI - Melatonin: possible mechanisms involved in its 'radioprotective' effect. AB - Peripheral blood samples were collected from four human volunteers before, and at 1 and 2 h after a single oral dose of 300 mg of melatonin. From each sample, separate aliquots of whole blood were exposed to 100-150 cGy gamma radiation in vitro to determine the extent of genetic damage. Irradiated lymphocytes from all volunteers which were collected after melatonin ingestion exhibited a significantly decreased extent of primary DNA damage and reduced frequencies of chromosomal aberrations and micronuclei, as compared with similarly irradiated cells collected before the oral dose of melatonin. The extent of the melatonin associated decrease in primary DNA damage was less than the corresponding decrease observed in the incidence of chromosomal aberrations and micronuclei; the latter assays required an additional post-irradiation incubation of the cells at 37+/-1 degreesC for 48 and 72 h, respectively. The possible mechanisms involved in the radioprotective influence of melatonin are discussed. PMID- 9729376 TI - Ciliary neurotrophic factor stimulates nuclear hypertrophy and increases the GFAP content of cultured astrocytes. AB - Studies using transgenic mice that overexpress ciliary neurotrophic factor (CNTF), direct injection of CNTF into brain parenchyma, and ectopic expression of CNTF by an adenoviral vector have demonstrated that CNTF activates astrocytes. Paradoxically, studies to date have failed to show an effect of CNTF on the expression of GFAP by cultured astrocytes. Therefore, the goal of this study was to use nuclear hypertrophy and GFAP expression as indices of glial activation to compare the responsiveness of forebrain type 1 and type 2 astrocytes to CNTF. As reported by others, CNTF did not increase GFAP in type 1 astrocytes; however, it rapidly increased their nuclear size by 20%. Nuclear hypertrophy was apparent within 4 h after CNTF exposure and persisted for at least 48 h. In contrast, type 2 astrocyte GFAP increased 2-fold over the course of 48 h of CNTF treatment. During this same treatment period type 2 astroglial nuclei enlarged by 25%. We conclude that CNTF stimulates both type 1 and type 2 astrocytes directly. Together with our in vivo studies (Levison et al., 1996: Exp. Neurol. 141: 256), these data support the concept that CNTF is responsible for many of the progressive astroglial changes that appear after CNS injury and disease. PMID- 9729377 TI - Taurine chloramine inhibits production of nitric oxide and prostaglandin E2 in activated C6 glioma cells by suppressing inducible nitric oxide synthase and cyclooxygenase-2 expression. AB - Taurine prevents tissue damage in various models of inflammation through a mechanism postulated to involve taurine monochloramine (Tau-Cl). Tau-Cl is formed through the action of a halide-dependent myeloperoxidase system associated with polymorphonuclear leukocytes (PMN), eosinophils, and basophils. Production of nitric oxide (NO), PGE2, and other proinflammatory mediators by activated macrophages is inhibited by Tau-Cl. Since glial cells may be activated to produce NO, PGE2 and other proinflammatory mediators, similar to macrophages, we examined the effects of Tau-Cl on the production of NO and PGE2 by rat C6 glioma cells. C6 cells were seeded to grow over 2-3 days to approximately 90% confluency before exposure to various concentrations of Tau-Cl in HBSS for 2 h (37 degreesC, 5% CO2). The HBSS was replaced, after washing the cells, with DMEM containing 4% fetal calf serum and activators (LPS, 10 microgram/ml; rat rIFN-gamma, 50 U/ml; and human rTNF-alpha, 50 ng/ml). Media content of NO2- and PGE2 was measured 48 h after activation and cell lysates were subjected to SDS-PAGE followed by Western blot analyses to determine the relative expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) proteins. Media accumulation of NO2- and PGE2 was inhibited by Tau-Cl in a concentration dependent manner and this was accompanied by decreased amounts of iNOS and COX-2 proteins in cell lysates. Additional experiments determined the effects of Tau-Cl on the kinetics of iNOS and COX-2 mRNA expression. Expression of iNOS mRNA was markedly inhibited in activated C6 cells that were previously exposed to Tau-Cl and this persisted for at least 24 h. In contrast, inhibition of COX-2 mRNA expression was only transiently reduced in Tau-Cl exposed cells during the first 4 h of activation and was relatively unimpaired thereafter (8-24 h). These results suggest that Tau Cl inhibits the transcriptional expression of the iNOS gene but inhibits expression of COX-2 protein by post-transcriptional mechanisms. PMID- 9729378 TI - Long term changes in brain cholinergic markers and nerve growth factor levels after partial immunolesion. AB - There are deficits in cholinergic basal forebrain neurons (CBFNs) in the aged brain and patients suffering Alzheimer's disease associated with a partial loss of the CBFNs. To mimic this partial loss and assess its long term effects on residual cholinergic activity and resultant target-derived nerve growth factor (NGF) levels, we produced a partial immunolesion to CBFNs with 192 IgG-saporin, an immunotoxin selectively taken up by p75NTR-bearing neurons. We measured two cholinergic markers, choline acetyltransferase (ChAT) and acetylcholinesterase (AChE) activity, and NGF protein levels at 10 days, 1, 6 and 12 months postlesion. There were no significant changes in the cholinergic markers and the NGF protein levels in the sham-treated animal controls during the one year experiment. Ten days after 192 IgG-saporin treatment, ChAT activity decreased to 35-50% of controls in the olfactory bulb, hippocampus, and cortex. There was a minor but significant recovery of ChAT activity one year after the immunolesion in the hippocampus. Changes in AChE activity mirrored the ChAT changes but were less robust. There were transient increases in NGF protein levels in the hippocampus and cortex that returned to basal levels at 6 months and 12 months postlesion, respectively. In summary, partial immunolesions resulted in partial region-specific and time-dependent recoveries of cholinergic activity in the target areas of the basal forebrain after a partial elimination of CBFNs and a return to basal levels of NGF protein consistent with the hypothesis that the remaining CBFNs compensated for losses of ChAT and NGF due to changes in cholinergic innervation of basal forebrain target areas. PMID- 9729379 TI - Chemically induced aneuploidy: investigations into chromosome specific effects in mitosis. AB - Genotoxicity studies of aneuploidy may potentially produce different results depending upon the chromosome selected for analysis if chromosome-specific sensitivities to chemical exposure exist. Any chromosome specificity characteristics that predispose to aneuploidy might interact with environmental exposures in additional different ways related to the mechanism of aneuploidy induction. Thus, we have undertaken an investigation of chromosome-specific effects using morphologically distinct chromosomes in a hybrid cell line. We were able to identify eight different chromosomes simultaneously by dual colour FISH analysis in controls and in cells exposed to a range of griseofulvin concentrations. Certain chromosomes were more frequently involved in aneuploidy, but no simple relationship between chromosome organisation and sensitivity emerged apart from the over-representation of the alien human chromosome. Aneuploidy was detected at higher frequencies in interphase cells compared with metaphase cells. Overall the data indicate that chemically induced aneuploidy may be detected for a variety of chromosomes and cell types using both interphase and metaphase protocols. However, the data obtained should be used with care in the hazard evaluation of chemical aneugens. PMID- 9729380 TI - High PCO does not alter pHi, but raises [Ca2+]i in cultured rat carotid body glomus cells in the absence and presence of CdC12. AB - We measured the effect of high PCO (500-550 Torr) on the pHi and [Ca2+]i in cultured glomus cells of adult rat carotid body (CB) as a test of the two models currently proposed for the mechanism of CB chemoreception. The metabolic model postulates that the rise in glomus cell [Ca2+]i, the initiating reaction in the signalling pathway leading to chemosensory neural discharge, is due to [Ca2+] release from intracellular Ca2+ stores. The membrane potential model postulates that the rise in [Ca2+]i comes from influx of extracellular Ca2+ through voltage dependent Ca2+ channels (VDCC) of the L-type. High PCO did not change pHi at PO2 of 120-135 Torr, showing that CO-induced changes in [Ca2+]i are not due to changes in pHi. High PCO caused a highly significant rise in [Ca2+]i from 90+/-12 nM to 675+/-65 nM, both in the absence and in the presence of 200 microM CdCl2, a potent blocker of L-type VDCCs. This result is fully consistent with release of Ca2+ from glomus cell intracellular stores according to metabolic model, but inconsistent with influx of extracellular Ca2+ through VDCCs according to the membrane potential model. PMID- 9729381 TI - Desensitization and enhancement of neurotensin/neuromedin N mRNA responses in subsets of rat caudate-putamen neurons following multiple administrations of haloperidol. AB - Striatal neurons that respond to blockade of dopamine receptors with altered expression of neurotensin/neuromedin N mRNA were examined. Injections of haloperidol were given to rats at four or 24 h and both four and 24 h prior to sacrifice. Pair-matched controls were injected with equivalent volumes of vehicle at either 4 or 24 h prior to sacrifice. Sections of striatum were processed non isotopically with a cRNA neurotensin/neuromedin N probe. Massive numbers of neurons exhibited hybridization in the lateral and dorsolateral caudate-putamen at 4 h. At 24 h, hybridized neurons were few in lateral and dorsolateral parts of the caudate-putamen, but more numerous in the dorsomedial and ventrolateral caudate-putamen than in controls. A second injection of haloperidol 4 h prior to sacrifice enhanced the dorsomedial/ventrolateral response, but failed to elicit substantial numbers of lateral and dorsolateral hybrids, as were observed at 4 h after one injection. Resistance of neurotensin expression to a second injection of haloperidol was selective for the lateral and dorsolateral parts of the caudate-putamen and may reflect residual blockade by haloperidol or altered DA receptors or second messengers. Sections subjected to immunohistochemical processing for neurotensin peptide and in situ hybridization with the neurotensin/neuromedin N mRNA probe exhibited numerous neurons in the dorsomedial and ventrolateral quadrants of the caudate-putamen that were double-labeled with immunoperoxidase and hybridization signals. This suggests that peptide synthesis, as opposed to decreased release of peptide, has a role in the accumulation of neurotensin immunoreactivity by dorsomedial and ventrolateral striatal neurons. PMID- 9729382 TI - Modulation of the formalin-induced nociceptive response by diabetes: possible involvement of protein kinase C. AB - Injection of formalin into the hind paw of mice produced a biphasic nociceptive response consisting of immediate (first-phase) and tonic (second-phase) components. Although the duration of the first-phase response was significantly longer in diabetic mice than in nondiabetic mice, the second phase was significantly shorter in diabetic mice. The first-phase response was dose dependently and significantly reduced by pretreatment with calphostin C (0.3 to 3 pmol, i.t.), a specific protein kinase C inhibitor, in diabetic mice. The second phase response was markedly increased when diabetic mice were pretreated with calphostin C. However, calphostin C (3 nmol, i. t.) had no significant effect on either the first-phase or second-phase response in nondiabetic mice. On the other hand, pretreatment with phorbol-12,13-dibutyrate (50 pmol, i.t.), a protein kinase C activator, significantly enhanced the first-phase response in nondiabetic mice. These results suggest that the change in the formalin-induced nociceptive response in diabetic mice may be due, at least in part, to the modification of nociceptive transmission in the spinal cord by the activation of protein kinase C. PMID- 9729383 TI - MAP kinase phosphatase 1 is expressed and enhanced by FK506 in surviving mamillary, but not degenerating nigral neurons following axotomy. AB - The MAP kinase phosphatase 1 (MKP-1), a dual serine-threonine phosphatase, inactivates the MAP kinases ERK and JNK/SAPK which are involved in neuronal survival and neuronal cell death following injury and degenerative stimuli. We have studied by immunocytochemistry whether regulation of MKP-1 is part of the cell-body response following nerve fiber transection. The expression of MKP-1 was investigated in axotomized neurons of the corpus mamillaris (CMm) and substantia nigra pars compacta (SNC) following transection of the mamillo-thalamic tract (MT) and the medial forebrain bundle (MFB), respectively. In contrast to the surviving CMm neurons, the vast majority of SNC neurons undergoes cell death following axotomy. MKP-1 immunoreactivity which is absent in untreated adult rats, appeared in CMm neurons 24 h following MT transection, reached a maximum after 2 days and persisted in a substantial proportion of CMm neurons until 20 days, the end of observation period. In contradistinction, MKP-1 could not be detected in the SNC neurons. MKP-1 immunoreactivity was virtually restricted to the nuclei of neurons. Subcutaneous injection of the immunosuppressant FK506 that protects axotomized SNC neurons against neuronal cell death, enhanced the expression of MKP-1 in CMm, but failed to do so in SNC neurons. The selective expression of MKP-1 in CMm is the first finding on a different regulation of components in the stress kinase signal pathway in surviving vs. degenerating axotomized neurons. PMID- 9729384 TI - Involvement of telomeric sequences in chromosomal aberrations. AB - The genomes of higher eukaryotes are not homogeneous in terms of structure or function. Many examples of chromosomal regions particularly prone to involvement in aberrations have been reported. The molecular structures of some of these regions have now been determined, most notably the folate-sensitive fragile sites and FRA16B-a distamycin A-sensitive fragile site. In addition, a number of cytological studies suggest that telomeric sequences can in some circumstances be involved in chromosomal aberrations more frequently than expected. Here, the roles of telomeric DNA sequences, both terminal and interstitial, and telomerase in chromosomal aberration formation are reviewed. PMID- 9729385 TI - Alterations of hypothalamic catecholamines in the newborn offspring of gestational diabetic mother rats. AB - Catecholamines are essential organizers of the developing brain. Throughout life, they are involved, e.g., in the regulation of body weight and metabolism by specific hypothalamic nuclei, which are suggested to be highly vulnerable to maternal gestational hyperglycemia. By application of streptozotocin (30 mg/kg, i.p.) gestational diabetes (GD) was induced in female rats. On the 1st day of life, male GD offspring were underweight (P<0.05) and hyperglycemic (P<0.05), while on the 21st day of life decreased body weight (P<0.001) and elevated pancreatic insulin (P<0.01) were observed. Using HPLC with electrochemical detection, hypothalamic catecholamines were determined in the newborns, and quantitative immunocytochemistry for tyrosine hydroxylase (TH) was performed. At birth, a tendency towards increased levels of norepinephrine (NE) and dopamine (DA) in the whole hypothalami of GD offspring was observed. In the 21-day-old offspring of GD mothers, NE was significantly increased in the ventromedial hypothalamic nucleus (VMN; P<0.05) and the lateral hypothalamic area (LHA; P<0.05), while DA was significantly elevated in the paraventricular hypothalamic nucleus (PVN; P<0.05) and the LHA (P<0.05). The NE/DA-ratio was found to be decreased in the PVN of GD offspring (P<0.01). Moreover, numerical density of TH positive neurons was clearly increased within the parvocellular division of the PVN (P<0.0001) as well as in the periventricular hypothalamic area (PER; P<0.05). These data suggest specific alterations of catecholaminergic systems within hypothalamic regulators of body weight and metabolism during early development in the offspring of gestational diabetic mother rats. PMID- 9729386 TI - Endomorphins decrease heart rate and blood pressure possibly by activating vagal afferents in anesthetized rats. AB - Endomorphin 1 (10, 30, 100 nmol/kg) administered intravenously (i.v. ) to urethane-anesthetized rats consistently and dose-dependently lowered heart rate (HR) and mean arterial pressure (MAP); the decrease in blood pressure recovered faster as compared to the HR. The effects of endomorphin 2 were qualitatively similar. Naloxone (2 mg/kg, i.v.) completely antagonized the bradycardia and hypotension caused by endomorphin 1. Pretreatment of the rats with atropine methylnitrate, atropine sulfate (2 mg/kg, i.v.) or bilateral vagotomy nearly abolished the bradycardia and attenuated the hypotensive effect of endomorphin 1. Our studies suggest that the bradycardia effect following systemic administration of the new opioid peptide may be explained by activation of vagal afferents and the hypotensive effect may be secondary to a reduction of cardiac output and/or a direct vasodilation. PMID- 9729387 TI - Induction of telomerase activity by in vivo X-irradiation of mouse splenocytes and its possible role in chromosome healing. AB - Telomeres serve as protective caps for the chromosome ends. They are one of the functional elements required for the stable transmission of eukaryotic chromosomes. Telomerase, a ribonucleoprotein, stabilises the telomere length by adding telomere repeats on to chromosome ends. Telomeres and telomerase can play a role in the formation of chromosome aberrations and especially in healing of the chromosome or chromatid breaks produced by radiation-induced DNA damage. Telomerase-independent processes also appear to be capable of capping broken chromosome ends. We have studied the expression of telomerase, telomere status and chromosome rearrangements in mouse splenocytes following different doses (0.5, 1.0, 2.0 or 3.0 Gy) of X-irradiation in vivo up to 224 days post-exposure. A dose-dependent increase in telomerase activity up to 2 Gy X-ray exposure was observed immediately after irradiation. The increased enzyme activity was detected even up to day 224 post-irradiation, the last time point studied, especially at higher doses (2 Gy and 3 Gy). A significant difference in average telomere length, measured by quantitative fluorescence in situ hybridisation (Q FISH) on metaphase chromosomes, noticed immediately after irradiation indicates terminal deletion or altered telomere chromatin. However, telomere length was not statistically significant from the control at the later time points studied. Presence of telomere repeats at the chromosomal breakage sites revealed by FISH with peptide nucleic acid (PNA) telomeric probe indicates a possible role of telomerase-dependent or independent processes in chromosome healing and telomere capture in mammalian cells. We found that approximately 25 to 50% of the newly formed telomeres at the breakage sites are in the range of 200 bp to 1 kb, which might suggest that these repeats could have been added by telomerase which showed a corresponding increase following irradiation. PMID- 9729388 TI - Testosterone as well as estrogen increases serotonin2A receptor mRNA and binding site densities in the male rat brain. AB - Our previous findings in female rats suggest that the potent effects of sex steroids on mood and mental state may be mediated, in part, by the effect of estrogen on the 5-hydroxytryptamine2A receptor (5-HT2AR) in brain. The aim of the present study was to determine the effect of acute (approximately 32h) sex steroid manipulation on central 5-HT2AR in the adult male Wistar rat. Castration (under halothane anesthesia) decreased while testosterone or estrogen, but not 5alpha-dihydrotestosterone (5alpha-DHT), increased significantly the 5-HT2AR mRNA content in dorsal raphe nucleus and the density of 5-HT2AR binding sites in frontal, cingulate and primary olfactory cortex and nucleus accumbens. The lack of effect of 5alpha-DHT, a potent androgen which cannot be converted to estrogen, suggests that the action of testosterone depends upon its conversion to estrogen by aromatase. This may also explain why estrogen, but not testosterone or 5alpha DHT, increased the density of 5-HT2AR binding sites in the caudate-putamen, a brain region where aromatase is scarce. These findings are discussed in relation to the possible role of the 5-HT2AR in depression, schizophrenia and Alzheimer's Disease. PMID- 9729389 TI - Differential effect of painful heterotopic stimulation on capsaicin-induced pain and allodynia. AB - Painful heterotopic stimulation (HTS) may inhibit experimental and clinical pain, an effect known as diffuse noxious inhibitory control (DNIC). This study examined the effect of painful HTS on capsaicin-induced pain intensity, brush-evoked pain intensity and area of brush-evoked pain in humans. Immersion of the foot into painful cold water significantly reduced capsaicin-induced pain intensity and brush-evoked pain intensity in the contralateral forearm, but did not change area of brush-evoked pain. The observed differential effect on the magnitude of pain and hyperalgesia on the one hand and area of hyperalgesia on the other suggests that the DNIC effect on spinal activity is selective and not general. PMID- 9729391 TI - Measuring parallel fiber length in the rat cerebellum. AB - A new approach was devised to obtain the mean length of parallel fibers in Golgi sections of the rat cerebellum. This method was based on the principle that within a given region of the cerebellum the mean length of parallel fibers should be inversely proportional to the likelihood of spotting the ends of parallel fibers. An experimental protocol was designed based on the statistical relationship between the mean fiber length and the number of the ends of parallel fibers for a given total length of parallel fibers examined. Our methodology was simple and could have advantages over other existing methods. PMID- 9729390 TI - Morphine microinjected into the nucleus raphe magnus does not block the activity of spinal trigeminal nucleus oralis convergent neurons in the rat. AB - This study investigated the effects of morphine microinjection into the nucleus raphe magnus (RMg) on electrically evoked C-fiber activities of convergent neurons in the spinal trigeminal nucleus oralis (Sp5O), in halothane-anesthetized rats. Although the neurons could be depressed by systemic morphine (6 mg/kg, i.v.) in a naloxone-reversible fashion, morphine microinjected into the RMg (2. 5 microgram or 5 microgram) neither depressed their C-fiber-evoked responses, nor the diffuse noxious inhibitory controls acting on them. It is concluded that the RMg is not involved in reinforcing descending inhibitory controls that are tonic or triggered by noxious stimuli acting on Sp5O convergent neurons. PMID- 9729392 TI - Expression of NMDA receptor subunit mRNA after MK-801 treatment in neonatal rats. AB - Although NMDA receptor antagonists are neuroprotective when delivered in conjunction with NMDA, supersensitivity to NMDA-mediated injury follows dizocilpine (MK-801) administration in neonatal rats. An increase in NMDA sensitive [3H]-glutamate binding accompanies the increase in vulnerability to excitotoxic injury. The present study tests the hypothesis that MK-801 may alter gene expression for the NMDA receptor subunits. Quantitative in situ hybridization histochemistry was used to evaluate the expression of NMDA receptor subunits NR1 and NR2A-D in neonatal rats, 2 to 4 h after treatment with MK-801. Increased mRNA for multiple NMDA receptor subunits was observed in cerebral cortex, striatum and hippocampus. The percent increase in NR2A mRNA was larger than the percent change in NR1, NR2B or NR2D. A small increase in mRNA for the metabotropic glutamate receptor mGluR5 was also observed after MK-801 treatment. These results indicate that gene expression for NMDA receptor subunits in the developing brain is rapidly altered after antagonist exposure. Increased expression of excitatory amino acid receptor subunit mRNA may contribute to the enhanced vulnerability to excitotoxic injury that has been observed after MK-801 treatment. PMID- 9729393 TI - 5-Hydroxytryptaminergic receptor agonists: effects on neuropeptide Y potentiation of feeding and respiratory quotient. AB - The objective of the present report was to characterize further the potential interactive effects of NPY and 5-HT on feeding and whole-body calorimetry. Specifically, several experiments examined the impact of various 5-HT receptor agonists on NPY stimulated eating and alterations in respiratory quotient (RQ). This included the 5-HT1A/1B receptor agonist RU 24969, the 5-HT1B/2C agonist TFMPP and the 5-HT2A/2C agonist DOI. In feeding tests conducted at the onset of the dark cycle, RU 24969, TFMPP and DOI were administered 5 min prior to PVN injection of NPY and food intake was measured 1 h postinjection. The metabolic effects of NPY following similar pretreatment were monitored using an open circuit calorimeter measuring the volume of oxygen consumed (VO2), carbon dioxide produced (VCO2) and RQ (VCO2/VO2). PVN injection of NPY (50-100 pmol) potentiated feeding and evoked reliable increases in RQ. DOI (5-20 nmol), but not RU 24969 (5 20 nmol) or TFMPP (10-40 nmol), antagonized NPY induced eating and blocked the peptide's effects on RQ. These findings suggest that 5-HT2A receptors within the PVN modulate NPY's effect on feeding and energy substrate utilization at the start of the nocturnal period. PMID- 9729394 TI - NMDA receptor subunits in the postsynaptic density of rat brain: expression and phosphorylation by endogenous protein kinases. AB - N-methyl-D-aspartate (NMDA) receptors (NRs) play critical roles in diverse synaptic processes in the brain. However, subcellular distribution, spatiotemporal expression and regulation of NR subunits in brain synapses are unknown. We report that NR1 and NR2A-2C subunits are all enriched in the postsynaptic density (PSD), which plays critical roles in trophin-mediated synaptic plasticity. Significant expression of NRs was observed the first two weeks after birth, during synaptogenesis, and in adulthood. Functional diversity of NRs, resulting from heterogeneous composition, was supported by the finding that different NR2 subunits were associated in a region-specific manner with NR1. Phosphorylation of NR1, a key subunit of the NMDA receptor-channel complex, was significantly enhanced by activators of calmodulin (CaM) kinases (CKs) or protein kinase C (PKC), but not by those of PKA. Co-immunoprecipitation studies revealed that NR1 was physically associated with functionally active PKCgamma and the major PSD protein (mPSDp) through noncovalent interactions. Our results suggest that NMDA receptors play roles in postsynaptic mechanisms in a subunit-, composition-, brain region- and developmental-specific manner. Our findings also indicate that the PSD is a coherent functional unit containing protein kinases that potentially regulate NMDA receptor function via phosphorylation. PMID- 9729395 TI - Telomere length and telomere-centromere relationships? AB - The quantitative fluorescence in situ hybridization (Q-FISH) technique enables an accurate estimate of individual telomere lengths, a possibility beyond the resolution of conventional techniques. So far, Q-FISH has been used for the estimate of individual telomere lengths in human, mouse and Chinese hamster chromosomes. This analysis revealed large variations in the size of individual telomeres and a specific intra-chromosomal distribution of telomere lengths; telomeres closer to centromeres appear to be shorter than their counterparts more distant from centromeres. This observation suggests that individual telomere length may be affected by centromere position, a possibility consistent with the theory of chromosome field postulated more than 40 years ago by Lima-de-Faria. The link between the theory of chromosome field and the role of telomere centromere relationships in the regulation of telomere length is discussed in this article. PMID- 9729396 TI - Melatonin inhibits spontaneous and VIP-induced vasopressin release from suprachiasmatic neurons. AB - We have studied melatonin effects on vasopressin release from dispersed cells of the rat suprachiasmatic nuclei (SCN). The release follows a circadian rhythm peaking during the day and decreasing at night. Melatonin inhibits the spontaneous increase and accelerates the decrease of vasopressin release. Melatonin also inhibits vasopressin release induced by vasoactive intestinal peptide (EC50=0.4 nM). The inhibition of vasopressin release correlates with the known inhibitory effect of melatonin on spontaneous neuronal activity in SCN. PMID- 9729397 TI - Changes in blood-brain barrier permeability associated with insertion of brain cannulas and microdialysis probes. AB - Blood-brain barrier (BBB) transcapillary transport was studied after insertion of cannulas and microdialysis probes into the brains of three groups of rats. Quantitative autoradiography was used to measure changes in BBB permeability around the insertion site. In the first group, BBB function was measured with 14C sucrose at times from immediately, and up to 28 days, after insertion of a microdialysis probe. BBB function was disrupted biphasically: a 19-fold increase in the influx constant (K1) of sucrose occurred immediately after insertion with a second 17-fold increase at 2 days, followed by a slow decline to 5 times normal values at 28 days. In the second group, 14C-dextran (70 kDa) was used to measure BBB transcapillary transport; K1 was increased 90-fold after probe insertion. In the 3rd group, 14C-AIB (alpha-aminoisobutyric acid) was used to evaluate BBB transport after insertion of a 27 gauge cannula, which was used to infuse 1 microliter of saline over 5 min. The K1 of AIB was increased 25 times control values. We conclude that BBB transcapillary transport function is disturbed in response to insertion of brain cannulas and/or microdialysis probes, that BBB dysfunction is maximal at the cannula or probe tip, varies with time after insertion, may persist for at least 28 days after insertion, and occurs over a wide molecular range of solutes. These results suggest caution when using microdialysis as a method to study normal BBB function, and suggest that microdialysis may overestimate the rate of transfer into and out of the brain. PMID- 9729398 TI - Posttreatment with low molecular weight heparin reduces brain edema and infarct volume in rats subjected to thrombotic middle cerebral artery occlusion. AB - Low molecular weight heparin (LMWH) has similar efficacy to unfractionated heparin with less hemorrhagic complications. We studied the neuroprotective effect of LMWH on a rat model of focal-ischemia. Our results revealed that treatment with LMWH at 1 and 3 h following thrombotic MCA occlusion reduced brain edema and infarct size and improved clinical outcome. Treatment with LMWH initiated at 6 h after thrombin injection only partially ameliorated brain damage. PMID- 9729399 TI - Parvalbumin immunoreactivity during the development of the cerebellum of the rainbow trout. AB - The distribution of parvalbumin immunoreactivity in the developing cerebellum of the rainbow trout was studied by using a specific monoclonal antibody and the avidin-biotin peroxidase method. Parvalbumin immunoreactivity was absent during the embryonic development of the cerebellum. The first immunoreactive elements, identified by their localization and posterior morphological evolution as immature Purkinje cells, appeared at 6 days posthatching in the presumptive corpus cerebelli and lobus vestibulolateralis. The labeling extended throughout the cerebellum following a caudorostral gradient, and in 21 days alevins, parvalbumin immunoreactive Purkinje cells were also observed in the valvula cerebelli. The appearance of parvalbumin-immunostaining in the Purkinje cells was not simultaneous; the labeling was observed initially in the cell body, extending gradually to the dendritic branches and finally to the axon. From 1 year onwards, parvalbumin immunoreactive terminal puncta from the Purkinje cell axons were observed surrounding the cell bodies of eurydendroid cells, that were parvalbumin immunonegative in all developmental stages studied. The spatio-temporal pattern of parvalbumin immunoreactivity in the rainbow trout cerebellum is different to previous observations in the cerebellum of amniotes. PMID- 9729400 TI - Neurons in the hypothalamic paraventricular nucleus that project to the rostral ventrolateral medulla are not activated by hypotension. AB - The retrogradely-transported tracer, rhodamine-tagged microspheres was injected into the pressor region of the rostral ventrolateral medulla (RVLM) to enable detection of paraventricular neurons in the hypothalamus that project to the RVLM. The protein, Fos, was detected immunohistochemically and used to highlight neurons that were activated by hypotension (-16+/-5 mmHg) induced by diazoxide (30 mg/kg s.c.). Compared to controls, Fos production was increased by three-fold in the parvocellular paraventricular nucleus but there was no significant increase in the number of retrogradely-labelled cells that expressed Fos. The results suggest paraventricular nucleus (PVN) neurons projecting to the RVLM are not activated by hypotension. PMID- 9729401 TI - Characterization and chromosomal mapping of two pseudogenes of the mouse Pafaha/Lis1 gene: retrointegration hotspots in the mouse genome. AB - Isolated lissencephaly sequence and Miller-Dieker syndrome are related neurodevelopmental disorders caused by defects of the LIS1 gene encoding the alpha subunit of intracellular platelet-activating factor acetylhydrolase. In addition to the ortholog of the human LIS1 gene (Pafaha/Lis1), the mouse genome contains two more homologs. In order to characterize the new members of this gene family, we isolated both Pafaha/Lis1-related genes (Pafaha-ps1 and Pafaha-ps2) from a mouse genomic library. Pafaha-ps1 and Pafaha-ps2 are processed pseudogenes formed by the retroinsertion of 5'-truncated Pafaha/Lis1 cDNAs. Sequence analysis revealed a striking accumulation of retroelements at both loci, identifying two retroinsertion hotspots in the mouse genome. The recognition of tRNA genes flanking Pafaha-ps1 provides an example for the potential association of RNA polymerase III transcription and retroinsertion in mammals. Linkage mapping placed Pafaha-ps1 and Pafaha-ps2 to distal chromosome (Chr) 3 and proximal Chr 7, respectively. Our results indicate that only one of the three LIS1-related mouse loci (Pafaha/Lis1) is functional, in contrast with two closely related functional genes (LIS1 and LIS2) reported in humans. 1998 Elsevier Science B.V. PMID- 9729402 TI - Differential interaction between 5-HT3 receptors and GABAergic neurons inhibiting acetylcholine release in rat entorhinal cortex slices. AB - The 5-HT3 receptor antagonists, ondansetron, MDL 72222 and granisetron (0.01-1 microM), produced a concentration-dependent increase of K+-evoked [3H]ACh efflux in slices from rat entorhinal cortex preloaded with [3H]choline. Bicuculline and flumazenil, antagonists at different sites of the GABAA receptor, also enhanced [3H]ACh efflux. While the ACh releasing effect of ondansetron was markedly potentiated, in a TTX-sensitive manner, by bicuculline, the effects of MDL 72222 and granisetron were not significantly modified. A qualitatively identical interaction was found by using flumazenil, a GABAA antagonist at the benzodiazepine recognition site, in combination with the 5-HT3 receptor antagonists. The potentiation by the GABAA antagonists of [3H]ACh efflux was also observed in a superfusion medium deficient in Cl-. The nonspecific K+-channel blockers TEA and Ba2+ also increased K+-evoked [3H]ACh efflux in this preparation but the releasing effect was not modified by bicuculline. The results support the functional interaction of ondansetron with GABAergic interneurons in the rat entorhinal cortex, GABA-independent mechanisms may however be involved in the regulation of cortical cholinergic function by other 5-HT3 receptor antagonists. PMID- 9729403 TI - Mosaic distribution of chondroitin and keratan sulphate in the developing rat striatum: possible involvement of proteoglycans in the organization of the nigrostriatal system. AB - The striatum of the mammalian basal ganglia is composed of two neurochemically distinct compartments termed patches and matrix that contribute overall to a mosaic organization. Glycosaminoglycans (GAGs), the sugar moieties of proteoglycans, provide specific spatio-temporal guidance cues during the development of several functional neural systems. However, their distribution within the nigrostriatal system has not been investigated yet. Here, the immunohistochemical distributions of unsulphated (C0S), 4-sulphated (C4S) and 6 sulphated chondroitin (C6S) and keratan sulphate (KS) were examined in the developing neostriatum of rat and compared with the distribution of dopaminergic terminals. All the chondroitin sulphate (CS) isomers are homogeneously expressed in the embryonic striatum. After birth, C0S and C6S reveal the striatal mosaic in being preferentially expressed within the matrix compartment and in boundaries around patches whereas the C4S epitope is present in both compartments, with a slight patchy distribution. KS expression is detected first in the patches during the early postnatal period and subsequently only in the matrix compartment. All these GAG expressions disappear as the brain matures except for C4S which remains high throughout adult life. Furthermore, studies within the developing medial forebrain bundle reveal that CS isomers, but not KS, are expressed in and around the dopamine axonal tract but show similar developmental patterns of distribution which do not appear to be specifically associated with the nigrostriatal pathway. These results suggest a possible implication of proteoglycans during the development of the striatum and may be useful for understanding the complex cellular and molecular interactions in degeneration and plasticity of the nigrostriatal circuit in Parkinson's disease. PMID- 9729404 TI - The pro-protein convertase PC1 is induced in the transected sciatic nerve and is present in cultured Schwann cells: comparison with PC5, furin and PC7, implication in pro-BDNF processing. AB - Injury of peripheral nerves induces expression of several pro-protein convertases (PCs) involved in processing of precursor proteins into their diverse active end products. In this study, the focus was on convertase PC1 which, although undetectable in control nerves, is strongly induced in injured nerves. High concentrations of PC1 mRNA of 9.0, 5.5, 3.0, 2.5 and 1.6 kb were observed on day 4 post-lesion in proximal and distal segments. By in situ hybridization PC1 mRNA was detected in most of endoneurial cells, which were further identified by immunocytochemistry as myelin 2', 3'-cyclic nucleotide 3'-phosphodiesterase containing Schwann cells. PC1 mRNA and protein were also present in cultured Schwann cells also containing convertases PC5, furin and PC7 as well as nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF). Mostly unprocessed pro-NGF of 35 kDa and pro-BDNF of 35 kDa were found on Western blotting of Schwann cells. Expression of exogenous neurotrophins by infection with vaccinia virus vector showed that mouse pro-NGF and rat pro-BDNF are cleaved intracellularly on smaller forms of 13.5 kDa NGF and 14 kDa BDNF. Infection experiments demonstrated that Schwann cells contain active processing enzymes. In conclusion, this work provides in vivo evidence of the presence of several PCs in the injured rat sciatic nerve and ex vivo in cultured Schwann cells. PMID- 9729405 TI - Genomic organization and promoter activity of the maize starch branching enzyme I gene. AB - Starch branching enzymes (SBE) which catalyse the formation of alpha-1,6-glucan linkages are of crucial importance for the quantity and quality of starch synthesized in plants. In maize (Zea mays L.), three SBE isoforms (SBEI, IIa and IIb) have been identified and shown to exhibit differential expression patterns. As a first step toward understanding the regulatory mechanisms controlling their expression, we isolated and sequenced a maize genomic DNA (-2190 to +5929) which contains the entire coding region of SBEI (Sbe1) as well as 5'-and 3'-flanking sequences. Using this clone, we established a complete genomic organization of the maize Sbe1 gene. The transcribed region consists of 14 exons and 13 introns, distributed over 5.7kb. A consensus TATA-box and a G-box containing a perfect palindromic sequence, CCACGTGG, were found in the 5'-flanking region. Genomic Southern blot analysis indicated that two Sbe1 genes with divergent 5'-flanking sequences exist in the maize genome, suggesting the possibility that they are differentially regulated. A chimeric construct containing the 5'-flanking region of Sbe1 (-2190 to +27) fused to the beta-glucuronidase gene (pKG101) showed promoter activity after it was introduced into maize endosperm suspension cells by particle bombardment. 1998 Elsevier Science B.V. PMID- 9729406 TI - Age- and region-specific expressions of the messenger RNAs encoding for steroidogenic enzymes p450scc, P450c17 and 3beta-HSD in the postnatal rat brain. AB - Neurosteroids are now known to be synthesized de novo in the nervous system through mechanisms at least partly independent of peripheral steroidogenic glands. In mammals, the presence of the cholesterol side-chain cleavage enzyme (cytochrome P450scc) and the enzyme 3beta-hydroxysteroid dehydrogenase/Delta5 Delta4-isomerase (3beta-HSD) has been well established in the brain, whereas limited information has been available on the enzyme 17alpha-hydroxylase/c17, 20 lyase (cytochrome P450c17), which converts pregnenolone to dehydroepiandrosterone, one of the most abundant neurosteroids. In addition, little is known regarding developmental changes in these steroidogenic enzymes during postnatal life. Thus, the pathway of neurosteroid formation in the brain is still incomplete. Therefore, we examined expressions of the messenger RNAs (mRNAs) encoding for three key enzymes, P450scc, P450c17 and 3beta-HSD, in the rat brain at different postnatal ages using RT-PCR analysis. The expression of P450scc mRNA was found throughout the brain at the same level, while the 3beta HSD mRNA expression was higher in the cerebellum and cerebrum than in other brain regions. The P450c17 mRNA was highly expressed in the mesencephalon. On the other hand, higher expressions of the cerebellar and cerebral 3beta-HSD mRNAs were observed only in neonatal life. In contrast, the expression of P450scc mRNA was relatively constant during neonatal life and in adulthood. A similar constant expression of the P450c17 mRNA was evident in the mesencephalon. Serial Southern hybridization in this study confirmed the specific mRNA expression corresponding to each enzyme. These results suggest that in the postnatal rat the expression of 3beta-HSD or P450c17 mRNA may be age- or region-dependent, unlike the P450scc mRNA expression. PMID- 9729407 TI - Neurons in the hypothalamic paraventricular nucleus send collaterals to the spinal cord and to the rostral ventrolateral medulla in the rat. AB - The hypothalamic paraventricular nucleus (PVN) projects to the rostral ventrolateral medulla (RVLM) and to the intermediolateral cell column (IML) of the spinal cord. The present study determined whether the same neurons can innervate both regions. In each rat, two retrogradely-transported tracers, microspheres tagged with fluorescein or rhodamine, were injected into the left lower thoracic/upper lumbar IML (fluorescein) and into the pressor region of the left RVLM (rhodamine). In the PVN over 90% of the neurons labelled with either tracer were found ipsilateral to the injection site. Double labelled cells averaged almost one-third of the spinally-projecting cells in four of the five animals. In the remaining animal, there were few double-labelled cells. The results suggest that a population of PVN neurons innervates both the lower thoracic/upper lumbar IML and the RVLM. PMID- 9729408 TI - The role of the complement system in traumatic brain injury. AB - A traumatic impact to the brain induces an intracranial inflammatory response, which consequently leads to the development of brain edema and delayed neuronal death. Evidence from experimental, clinical, and in vitro studies highlight an important role for the complement system in contributing to inflammation within the injured brain. The present review summarizes the current understanding of the mechanisms of complement-mediated secondary brain injury after head trauma. PMID- 9729409 TI - Lesions of glucose-responsive neurons impair synchronizing effects of calorie restriction in mice. AB - Calorie restriction can induce phase-advances of daily rhythms in rodents exposed to light-dark cycles. To test whether glucose-responsive neurons are involved in the synchronizing effects of calorie restriction, C57BL/6J mice were injected with gold-thioglucose (GTG; 0.6 g/kg) which damages glucose-responsive neurons, primarily located in the ventromedial hypothalamus. From the day of injection, GTG-treated and control mice received a hypocaloric diet (66% of ad libitum food intake) 2 h after lights on. When mice were transferred to constant darkness after 4 weeks and fed ad libitum, the onset of circadian rhythm of locomotor activity was phase-advanced by 1 h in control but not in GTG-treated mice. Therefore, glucose-responsive neurons in the ventromedial hypothalamus may play a role in the synchronizing effects of calorie restriction on circadian rhythmicity. PMID- 9729411 TI - Linkage of two distinct AT-rich minisatellites at multiple loci in the genome of Theileria parva. AB - Minisatellite tandem repeat elements are well known components of vertebrate genomes, but have not yet been extensively characterized in lower eukaryotes. We describe two unusual, AT-rich minisatellites of the protozoan parasite Theileria parva whose sequences are unrelated to the G/C-rich i minisatellite superfamily' of vertebrate and plant genomes. The T. parva tandem repeats, one with a conserved sequence T2-5ACACA (6-17 copies), and the other with a 6-bp core sequence of either ACTATA or TATACT associated with additional variable sequences in repeats of 10-17bp (3-7 copies), were closely linked at more than 20 sites in the T. parva genome, separated by 390, 510 and 660bp at three loci analysed in detail. Such linkage is without precedent in minisatellites so far analysed in other organisms. The minisatellite loci were widely dispersed on 13 out of 33 genomic SfiI fragments, on all four T. parva chromosomes and did not exhibit a telomeric bias in their distribution. Analysis of flanking sequences revealed no obvious conserved sequences between the five loci, or other multicopy repeat sequences outside the minisatellite regions. The T2-5 ACACA minisatellite was highly effective as a multilocus fingerprinting probe for discrimination of T. parva isolates. Analysis of two individual minisatellite loci revealed variation between the genomic DNAs of two T. parva isolates in the copy number of the constituent repeats within the array, similar to that typical of vertebrate minisatellites. 1998 Elsevier Science B.V. PMID- 9729410 TI - Expression of PACAP, and PACAP type 1 (PAC1) receptor mRNA during development of the mouse embryo. AB - Pituitary adenylate cyclase activating polypeptide (PACAP) and vasoactive intestinal polypeptide (VIP) have been reported to have a number of neurotrophic effects. We have examined the expression of mRNA for PACAP and PACAP type 1 (PAC1) receptor in the mouse embryo by in situ hybridization and the effects of PACAP and VIP on the growth of mouse embryos in vitro. Although we were unable to detect gross effects of either peptide on the growth rates of embryos maintained in culture, mRNAs for both PAC1 receptor and PACAP peptide were present in the nervous system from day 9.5 of embryonic development. PAC1 receptor mRNA was most abundant in the neural tube and the rhombencephalon and was present also in the dorsal root and trigeminal ganglia and the sympathetic chain. The distribution of mRNA for the PACAP peptide overlapped in part with that of the receptor, but was more extensively distributed in the rhombencephalon and in the developing hypothalamus. Within the neural tube, PAC1 receptor mRNA was located in the roof and floor plates, while the distribution of PACAP peptide mRNA was more complex, being located in two columns of cells in the ventromedial neural tube (consistent with the position of developing autonomic motor neurons) and in cells in the dorsolateral neural tube. These data are concordant with a role for PACAP or a related peptide in neural development. PMID- 9729412 TI - The effects of antisense to Gialpha2 on opioid agonist potency and Gialpha2 protein and mRNA abundance in the mouse. AB - In this study, mice received a single intracerebroventricular (i.c.v. ) injection of an antisense oligodeoxynucleotide (ODN) directed towards the mRNA of Gialpha2. Controls received a saline or a nonsense ODN injection. The subsequent effects on protein levels and mRNA of Gialpha2 were determined in mouse striatum, as well as, the effect on opioid ([d-Ala2, d-Leu5]-enkephalin; DADLE) inhibition of cyclic AMP (cAMP) formation in striatum and morphine analgesic potency. At 48 h after treatment, maximal inhibition (Emax) of cAMP formation was significantly reduced for the antisense group compared to controls. Antisense ODN treatment only changed the Emax and did not significantly alter the IC50s of the dose effect curves for inhibition of cAMP formation. Antisense ODN, but not nonsense ODN, significantly reduced morphine's analgesic potency by >2-fold, 48 h following treatment. Using a quantitative immunoblotting procedure, antisense treatment was shown to decrease striatal Gialpha2 protein 48 h after antisense injection, while there were no changes in protein levels at 2, 12 and 24 h. In contrast, no changes in Gialpha2 mRNA in mouse striatum were noted at any time after antisense treatment. Taken together, these data suggest that Gialpha2 mediates opioid-induced analgesia and opioid inhibition of cAMP production in the mouse. These data also suggest that antisense reduces target protein by a mechanism independent of changes in mRNA abundance. PMID- 9729413 TI - Self-sustaining status epilepticus after brief electrical stimulation of the perforant path. AB - We examined the duration of intermittent perforant path stimulation (PPS) needed to induce self-sustaining status epilepticus (SSSE) in rats. Seven-minute PPS did not induce SSSE. Some rats receiving 15 min and all animals after 30 min PPS developed SSSE that continued for hours. The animals killed 3 days after SSSE showed extensive neuronal damage. Those which were allowed to survive for 6 weeks after SSSE displayed spontaneous seizures. PMID- 9729414 TI - Spontaneous rhythm in c-Fos immunoreactivity in the dorsomedial part of the rat suprachiasmatic nucleus. AB - In rats maintained for 2 days in constant darkness, the suprachiasmatic nucleus exhibited a circadian rhythm in c-Fos immunoreactivity, with the maximum in the morning and trough during the subjective night. In contrast to the night-time photic c-Fos induction occurring in the ventrolateral part of the nucleus, the spontaneous rhythmic c-Fos induction in darkness occurred in the dorsomedial part and might indicate an elevated dorsomedial neuronal activity in the early subjective day. PMID- 9729415 TI - Characterization of the enzymatic domains in the modular polyketide synthase involved in rifamycin B biosynthesis by Amycolatopsis mediterranei. AB - Five clustered polyketide synthase (PKS) genes, rifA-rifE, involved in rifamycin (Rf) biosynthesis in Amycolatopsis mediterranei S699 have been cloned and sequenced (August, P.R. et al., 1998. Chem. Biol. 5, 69-79). The five multifunctional polypeptides constitute a type I modular PKS that contains ten modules, each responsible for a specific round of polyketide chain elongation. Sequence comparisons of the Rf PKS proteins with other prokaryotic modular PKSs elucidated the regions that have an important role in enzyme activity and specificity. The beta-ketoacyl:acyl carrier protein synthase (KS) domains show the highest degree of similarity between themselves (86-90%) and to other PKSs (78-85%) among all the constituent domains. Both malonyl-coenzyme A (MCoA) and methylmalonyl-coenzyme A (mMCoA) are substrates for chain elongation steps carried out by the Rf PKS. Since acyltransferase (AT) domains of modular PKSs can distinguish between these two substrates, comparison of the sequence of all ten AT domains of the Rf PKS with those found in the erythromycin (Er) (Donadio, S. and Katz, L., 1992. Gene 111, 51-60) and rapamycin (Rp) (Haydock, S. et al., 1995. FEBS Lett. 374, 246-248) PKSs revealed that the AT domains in module 2 of RifA and module 9 of RifE are specific for MCoA, whereas the other eight modules specify mMCoA. Dehydration of the beta-hydroxyacylthioester intermediates should occur during the reactions catalysed by module 4 of RifB and modules 9 and 10 of RifE, yet only the active site region of module 4 conforms closely to the dehydratase (DH) motifs in the Er and Rp PKSs. The DH domains of modules 9 and 10 diverge significantly from the consensus sequence defined by the Er and Rp PKSs, except for the active site His residues. Deletions in the DH active sites of module 1 in RifA and module 5 in RifB and in the N- and C-terminal regions of module 8 of RifD should inactivate these domains, and module 2 of RifA lacks a DH domain, all of which are consistent with the proposed biosynthesis of Rf. In contrast, module 6 of RifB and module 7 of RifC appear to contain intact DH domains even though DH activity is not apparently required in these modules. Module 2 of RifA lacks a beta-ketoacyl:acyl carrier protein reductase (KR) domain and the one in module 3 has an apparently inactive NADPH binding motif, similar to one found in the Er PKS, while the other eight KR domains of the Rf PKS should be functional. These observations are consistent with biosynthetic predictions. All the acyl carrier protein (ACP) domains, while clearly functional, nevertheless have active site signature sequences distinctive from those of the Er and Rp PKSs. Module 2 of RifA has only the core domains (KS, AT and ACP). The starter unit ligase (SUL) and ACP domains present in the N-terminus of RifA direct the selection and loading of the starter unit, 3-amino-5-hydroxybenzoic acid (AHBA), onto the PKS. AHBA is made by the products of several other genes in the Rf cluster through a variant of the shikimate pathway (August, P.R. et al., inter alia). RifF, produced by the gene immediately downstream of rifE, is thought to catalyse the intramolecular cyclization of the PKS product, thereby forming the ansamacrolide precursor of Rf B. 1998 Elsevier Science B.V. PMID- 9729416 TI - Postnatal development of signal generation in auditory thalamic neurons. AB - Using whole cell recording techniques, we distinguished immature from mature stages of development in auditory thalamic neurons of rats at ages P5 to P21. We compared voltage responses to injected currents and firing patterns of neurons in ventral partition of medial geniculate body (MGBv) in slices. Resting potential, input resistance and membrane time constant diminished to mature values between P5 and P14. Responses of young neurons to hyperpolarizing pulses showed delayed inward rectification; after P13, this was obscured by a rapid onset of another inward rectifier. All neurons possessed tetrodotoxin (TTX)-sensitive, depolarization-activated rectification, implying persistent Na+-current involvement. Despite a slightly higher voltage threshold for spiking, the current threshold was lower in younger neurons. Young neurons fired a short latency spike with afterhyperpolarization whereas older neurons exhibited a slow ramplike depolarization before tonic firing. Large currents caused continuous firing in all neurons. Before day P13, a high threshold Ca2+ spike (HTS) often was appended to action potentials. The low threshold Ca2+-spike (LTS) was too small in amplitude to evoke action potentials before P11 but produced a single spike at P12 and P13 and burst firing with HTS after P13. MGBv neurons have mature properties after P14, relevant for reactions to sound and the oscillations of slow-wave sleep. PMID- 9729417 TI - Lipopolysaccharide-induced expression of IP-10 mRNA in rat brain and in cultured rat astrocytes and microglia. AB - Using mRNA differential display technique, we have found a differentially expressed band in rat brain, designated HAP2G1, which was the strongest one induced in response to peripheral administration of lipopolysaccharide (LPS). Sequence analysis showed that HAP2G1 cDNA is the rat homologue of the human alpha chemokine IP-10. Using RT-PCR technique and in situ hybridization, we demonstrate that IP-10 mRNA was expressed only in brain tissue of rats treated with LPS and not in control brain tissue. Using semi-quantitative PCR, we found that both cultured astrocytes and microglia express IP-10 mRNA after treatment with LPS. LPS-induced IP-10 mRNA reached peak levels in rat brain and in cultured microglia at approximately 3 h after treatment with LPS. At 10 h, IP-10 mRNA was markedly decreased, and at 24 h it was low but still detectable by PCR or in situ hybridization. In contrast to unstimulated microglia, unstimulated astrocytes constitutively expressed IP-10 mRNA at a low level. Increased IP-10 expression could possibly be involved in the microglia response to inflammatory stimuli in vivo. PMID- 9729418 TI - Expression of iron transport proteins and excessive iron accumulation in the brain in neurodegenerative disorders. AB - New findings on the role of LfR (lactotransferrin receptor), MTf (melanotransferrin), CP (ceruloplasmin) and DCT1 (Divalent Cation Transporter) in brain iron transport, obtained during the past 3 years, are important advances in the fields of physiology and pathophysiology of brain iron metabolism. According to these findings, disruption in the expression of these proteins in the brain is probably one of the important causes of the altered brain iron metabolism in age related neurodegenerative diseases, including Parkinson's Disease, Alzheimer's disease, Huntington's disease and amyotrophic lateral sclerosis. Further studies on the involvement of LfR, MTf and DCT1 in iron uptake by and CP in iron egress from different types of brain cells as well as control mechanisms of expression of these proteins in the brain are critical for elucidating the causes of excessive accumulation of iron in the brain and neuronal death in neurodegenerative diseases. PMID- 9729419 TI - Central analgesic actions of loperamide following transient permeation of the blood brain barrier with Cereport (RMP-7). AB - The bradykinin analog, Cereport (RMP-7), was designed to increase permeability of the blood brain barrier (BBB). Over the past several years it has been developed primarily as a means of increasing permeability of the blood brain tumor barrier, where early evidence indicated a particularly robust and reliable effect. The present series of experiments were intended to determine whether Cereport might also be used to increase delivery of pharmacological agents across the normal (i.e., non-tumor) BBB. This was accomplished by testing the ability of Cereport to enhance delivery of the peripherally acting opiate agonist, loperamide, to the brain, as evidenced by induction of a centrally mediated analgesic effect. Intravenous administration of a combination of Cereport and loperamide produced a significant analgesic effect (2-fold increase in response times) when animals were tested on a hotplate apparatus. Loperamide alone did not produce analgesia. An analysis of the time course of analgesia revealed a graded onset of analgesia which peaked at 30 min, maintained asymptote at 60 min, and began to diminish by 120 min following Cereport and loperamide administration. Finally, the analgesic effects of combining Cereport and loperamide were completely blocked when animals were pre-treated with the opiate antagonist naloxone, demonstrating that the analgesia was mediated through opiate receptors. Collectively, these results suggest that Cereport was able to increase delivery of loperamide across the BBB, allowing it to gain access to opiate receptors in the CNS to produce a centrally mediated analgesic effect. They therefore provide clear evidence that safe and well-tolerated doses of Cereport can increase permeability of the normal (i.e., non-tumor) BBB. Moreover, they provide the first evidence of a pharmacological effect specifically enabled by controlled (i.e., receptor-mediated) modulation of the BBB. PMID- 9729421 TI - Intrauterine hypoxia-ischemia alters nitric oxide synthase expression and activity in fetal and neonatal rat brains. AB - The effects of intrauterine hypoxia-ischemia (HI) on nitric oxide synthase (NOS) activity and on expression of NOS isoforms were investigated in fetal and neonatal rat brains. Rat fetuses were subjected to either a 30-min intrauterine HI insult or a sham operation (SH) on gestational day 17 (G17). NOS activity in the homogenate of the rat brain was detectable on G17 and increased with age. NOS activity in the HI group was 20-30% higher than in the SH group from 6 to 48 h after the HI, but was 30% lower than in the SH group from postnatal day 8 to 14. Expression of the inducible NOS (iNOS) mRNA, as examined by RT-PCR, was increased as compared to the SH group from 6 to 24 h after the HI surgery. Expression of the constitutive neuronal NOS (nNOS) mRNA was reduced in the HI group from 24 h after the HI surgery up to postnatal day 14. Immunoblotting data have shown that alterations in NOS isoform protein expression caused by the intrauterine HI were consistent with the mRNA expression data. The overall results indicate that prenatal HI has long-lasting effects on function and expression of NOS in fetal and neonatal rat brains and that the altered NOS activity may be associated with prenatal HI-induced neurological abnormalities. PMID- 9729420 TI - Expression of trkB mRNA is altered in rat hippocampus after experimental brain trauma. AB - Recent investigations have shown that expression of mRNAs for the neurotrophins brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3) is differentially altered in the hippocampus following traumatic brain injury. In the present study, modulation of neurotrophin receptor expression was examined in the hippocampus in a rat model of traumatic brain injury using in situ hybridization. Messenger RNA for trkB, the high-affinity receptor for BDNF and neurotrophin-4 (NT-4), was increased between 3 and 6 h bilaterally in the dentate gyrus following a lateral fluid-percussion brain injury of moderate severity (2.0 2.1 atm). No time-dependent alterations were observed for trkB mRNA in hippocampal subfields CA1 and CA3. Levels of mRNA for trkC, the high-affinity receptor for NT-3, did not change in any region of the hippocampus. These data demonstrate that lateral fluid-percussion injury modulates expression of trkB mRNA in the hippocampus and support a role for BDNF/trkB signalling mechanisms in secondary events associated with traumatic brain injury. PMID- 9729422 TI - The new gene DmX from Drosophila melanogaster encodes a novel WD-repeat protein. AB - DmX is a novel gene from Drosophila melanogaster located on the X chromosome in region 5D5/6-E1. The molecular analysis of the genomic and cDNA sequences of DmX shows that the gene spans appr. 16kb and displays a mosaic structure with 15 exons. The 12kb long DmX transcript is present in Drosophila embryos, larvae and adults of both sexes. The open reading frame of DmX encodes a novel WD-repeat protein, containing at least 30 WD-repeat units. WD-repeat proteins contain a conserved motif of approximately 40 amino acids (aa), usually ending with the dipeptide Trp-Asp (WD). Homologues of the DmX gene exist in other dipteran species, in Caenorhabditis elegans and human, revealing that DmX is an evolutionarily well conserved gene. The inferred DMX amino acid sequence shows also limited, but significant similarity to a yeast ORF with unknown function. 1998 Elsevier Science B.V. PMID- 9729423 TI - Gene expression of the P2X receptors in the rat retina. AB - Molecular-biological methods were used to demonstrate the expression of six P2X receptor subunits (P2X1-P2X6) in retina and choroid. Despite the considerable evidence for signalling by extracellular nucleotides in other sensory systems, few studies have been undertaken in the eye. RT-PCR for the detection of P2X subunit mRNA in the rat of different postnatal developmental stages (P23-P210) revealed the presence of P2X2 and P2X4 mRNA in the retina and choroid; P2X3, and P2X5 were detected only in the retina. There was no evidence for P2X1 and P2X6 mRNA in the ocular tissue under investigation. Our data suggest that extracellular ATP may have influences on visual processing. PMID- 9729424 TI - Expression of mRNA for the elav-like neural-specific RNA binding protein, HuD, during nervous system development. AB - The expression of mRNA for the neuronal antigen HuD (Elavl4) associated with paraneoplastic encephalomyelitis and sensory neuronopathy was evaluated in the developing and adult rat nervous system. Using RNase protection assay and non radioactive in situ hybridization histochemistry HuD expression was shown to be expressed at high levels at the earliest time point observed (E15), but declined significantly during the first postnatal week to levels which were maintained into adulthood. In the adult, HuD expression became restricted primarily to large pyramidal-like neurons. Exceptions of note were many smaller neurons within a variety of thalamic nuclei. Expression of HuD was observed to be coincident with terminal differentiation of all neuronal structures evaluated regardless of the timing of their development, providing correlative evidence for a role in neuronal differentiation or the maintenance of neuronal phenotype. The marked restriction of HuD mRNA expression with maturity suggests that its functional role in adult neurons varies significantly throughout the CNS. PMID- 9729426 TI - Letter to the editor: reply. PMID- 9729425 TI - Trehalose-6P synthase is essential for trehalase activation triggered by glucose, nitrogen source or heat shock, but not by osmostress, in Schizosaccharomyces pombe. AB - Cells of Schizosaccharomyces pombe disrupted in the tps1+ gene, which encodes trehalose-6P synthase, were unable to increase trehalase activity in response to the addition of glucose or nitrogen source. Moreover, in contrast to normal cells, Deltatps1 cells did not increase trehalase activity by heat shock. Overexpression of tps1+ in cells devoid of trehalose-6P synthase restored the ability to increase trehalase after addition of nutrients or by heat shock. In glucose-repressed cells, which are normally refractory to the activation of trehalase by glucose, overexpression of tps1+ enabled the cells to increase trehalase activity upon addition of the sugar. Northern hybridisations were used to determine the level of mRNA for trehalase in normal and Deltatps1 cells. Transcription for trehalase was not significantly altered upon addition of glucose or nitrogen source, but increased markedly in heat-shocked cells even though trehalase activity remained unchanged in Deltatps1 cells. These findings provide evidence for a role of trehalose-6P synthase in the signalling pathway causing post-transcriptional activation of neutral trehalase induced by nutrients or heat shock. However, trehalase increased in Deltatps1 cells under hypertonic conditions suggesting the existence in Schiz. pombe of a distinct regulatory mechanism for enhancement of trehalase, specifically triggered by osmostress. PMID- 9729427 TI - Evidence of altered hypothalamic pro-opiomelanocortin/ neuropeptide Y mRNA expression in tubby mice. AB - The tubby mouse is characterized by an autosomal recessive mutation which results in the development of maturity-onset obesity and sensorineural hearing loss and retinal degeneration. Although the tubby mutation which leads to a splicing defect of the tub gene has been identified recently, the mechanism by which it causes the obesity syndrome has not been established. In this study, the potential dysfunction of several hypothalamic neuroendocrine pathways involved in the central regulation of energy metabolism was investigated in tubby mice. In comparison with the wild-type controls, a significant reduction (20%) of pro opiomelanocortin (POMC) mRNA expression was observed in the arcuate nucleus (ARC) of the mature, obese but not in the juvenile, non-obese tubby mice. Similarly, an age and body mass-dependent induction (about 30-fold) of neuropeptide Y (NPY) mRNA was observed in the dorsomedial (DMH) and ventromedial (VMH) hypothalamic nuclei of the tubby mice. However, NPY mRNA in the ARC was decreased by approximately 30 to 40% in both juvenile and mature tubby mice. The hypothalamic expression patterns of corticotropin releasing hormone (CRH) and the long form leptin receptor (OB-Rb) were not significantly altered in the mutant mice. These results suggest that the altered hypothalamic POMC and/or NPY functions may be important contributing factors for the development of obesity in this animal model. PMID- 9729429 TI - A novel isoform of the orphan nuclear receptor RORbeta is specifically expressed in pineal gland and retina. AB - RORbeta is a member of the nuclear hormone receptor superfamily whose ligand is unknown. Expression of RORbeta is confined to the central nervous system and its pattern suggests that this orphan nuclear receptor is implicated in the processing of sensory information and in circadian timing. In rats, RORbeta mRNA levels oscillate robustly in pineal gland and retina, displaying a 24h rhythm. Here we report the cloning of the cDNA of a novel isoform of RORbeta from rat pineal tissue. Expression of this isoform, called RORbeta2, is confined to pineal gland and retina and strongly increases at night. RORbeta2 shares common DNA- and putative ligand-binding domains with the canonical RORbeta (referred to as RORbeta1), but is characterized by a different amino-terminal domain. This structural difference renders RORbeta2 much more selectively binding to DNA than RORbeta1. Moreover, in contrast to RORbeta1, the novel isoform efficiently activates transcription also in non-neuronal cell lines. Thus, the two RORbeta isoforms are likely to regulate different sets of genes in different physiological contexts. 1998 Elsevier Science B.V. PMID- 9729430 TI - Conjugates of monoclonal antibodies with polyelectrolyte complexes-- an attempt to make an artificial chaperone. AB - Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from rabbit muscle is a tetrameric enzyme. Inactivation of GAPDH at 50 degreesC or in the presence of 4 M urea proceeds via formation of inactive dimers followed by their aggregation. Antibodies (clone 6C5) were selected which bind specifically inactive dimers but not native tetramers. The simplified model of chaperone action when the inactive misfolded forms are removed from the reaction media preventing aggregation was developed using antibodies in combination with polyelectrolyte complexes. The antibodies were coupled covalently to polyanionic component of the complex, poly(methacrylic) acid. The treatment of inactivated GAPDH with this conjugate followed by its precipitation after equimolar addition of polycation, poly-(N ethyl-4-vinylpyridinium) bromide, resulted in a significant increase in the specific activity of GAPDH. The restoration of specific activity was more complete in the experiments with lower GAPDH concentration and in the samples with lower inactivation degree, conditions where aggregation is less pronounced. Some aggregates are formed at high inactivation degree and high GAPDH concentration and observed as an increase in the solution turbidity. They could be removed by centrifugation. Antibody/polyelectrolyte complex treatment followed by centrifugation to remove insoluble aggregates resulted in nearly complete restoration of enzyme specific activity. PMID- 9729431 TI - Ontogeny of cation-Cl- cotransporter expression in rat neocortex. AB - Neuronal precursors and immature cortical neurons actively accumulate Cl- and as a consequence depolarize in response to GABAA receptor activation. With maturity, intracellular Cl- decreases resulting in a shift towards GABAA inhibition. These observations suggest that changes in expression of cation-Cl- cotransporters may have a significant role in the ontogeny of neuronal Cl- homeostasis. Using ribonuclease protection analysis and in situ hybridization we examined the developmental expression of all presently known members of the cation-Cl- cotransporter gene family in rat brain. Of the inwardly directed cotransporters, NKCC-1, NKCC-2, and NCC-1, only NKCC-1 was detected at significant levels in brain. NKCC-1 was expressed in neurons, appearing first in cortical plate but not in ventricular or subventricular zone. Expression levels peaked by the third postnatal week and were maintained into adulthood. The outwardly directed cotransporters, KCC-1 and KCC-2, demonstrated significantly different levels and time courses of expression. KCC-1 was expressed prenatally at very low levels which increased little over the course of development. In contrast, KCC-2 expression appeared perinatally and increased dramatically after the first week of postnatal life. Differential changes in expression of this gene family occurred during periods of critical shifts in chloride homeostasis and GABA response suggestive of a role in these processes. Furthermore the absence of expression of known inwardly directed cotransporters in Cl- accumulating neuroepithelia and lack of evidence for glial expression suggests that as yet unidentified members of this gene family may be involved in chloride homeostasis in immature neuronal precursors and neuroglia. PMID- 9729432 TI - Correlation of chi orientation with transcription indicates a fundamental relationship between recombination and transcription. AB - Cross-over hot-spot instigator (Chi) sequences (5'-GCTGGTGG-3') are abundant, strand-specific, sequences, which locally increase recombination in Escherichia coli. Located within G-rich 'recombination islands', Chi orientations correlate with the orientations both of DNA replication and of transcription. Consistent with evidence from eukaryotic systems for a fundamental relationship between recombination and transcription, we find for E. coli Chi sequences, and for Haemophilus influenzae Chi-like sequences, that orientations correlate better with transcription than with replication. Complying with Szybalski's transcription direction rule, open reading frames in these prokaryotes have purine-rich mRNA-synonymous DNA strands. Hence, the G-richness of 'recombination islands' may reflect their correspondence with 'transcriptional islands' (genes). Comparison of a natural with the corresponding shuffled sequence, indicates a base order-dependent island unit of approx. 1kb. 1998 Elsevier Science B.V. PMID- 9729433 TI - Sialyllactose occurs as free lactones in ovine colostrum. AB - Three sialyl oligosaccharide fractions were separated from ovine colostrum by gel filtration, anion exchange chromatography and normal-phase HPLC. They were characterized by 1H-NMR spectrometry as follows: Neu5Acalpha2-->3Galbeta1-->4Glc, Neu5Gcalpha2-->6Galbeta1-->4Glc and three forms of Neu5Gcalpha2-->3Galbeta1- >4Glc, namely Neu5Gcalpha2-->3Galbeta1-->4Glc itself, its lactone derivative between the carboxyl group of Neu5Gc and Gal OH-2 and another lactone derivative between the carboxyl group and Gal OH-4. In this study, Neu5Gc-lactose lactones, in their free form, have been isolated for the first time from any natural sources including milk or colostrum. PMID- 9729434 TI - Identification of immediate early genes during TPA-induced human myeloblastic leukemia ML-1 cell differentiation. AB - Human myeloblastic ML-1 can be induced to differentiate into monocytes/macrophages by 12-0-tetradecanoylphorbol-13-acetate (TPA). In order to understand the molecular mechanism regulating ML-1 cell differentiation, we focused on the characterization of immediate early genes activated by TPA using the mRNA differentiation display polymerase chain reaction (DD-PCR) and Northern analyses. A modified procedure, the reverse dot slot, was developed to confirm upregulated genes during the early stages of TPA-induced ML-1 cell differentiation. DNA sequencing analyses of 10 subcloned cDNA fragments, selected on the basis of the outcome of the reverse dot slot procedure, revealed that eight were derived from distinct genes. Among these clones, one was a novel gene (G07-5), another (A02-1) was highly homologous to the sequence of a fetal brain cDNA fragment, and the remaining six corresponded to jun-D, rantes, ssat, CD 14, ferritin heavy chain (fhc) and transposons Tn10-like transcript, respectively. Although these genes were all upregulated by TPA, the peak time of mRNA expression varied. jun-D, ssat and A02-1 expressions were superinduced in the presence of cycloheximide, which indicates that they belong to the immediate early gene family. On the other hand, TPA-induced rantes expression was not superinduced by cycloheximide, suggesting a protein synthesis-dependent process. As there are no previous reports of expression of these genes in TPA-induced ML-1 cells, little or no information is available concerning their function in mediating myeloblastic cell differentiation. Thus, this study illuminates new avenues of research for elucidating the function of genes regulating terminal differentiation of myeloid progenitors. 1998 Elsevier Science B.V. PMID- 9729435 TI - Microwave dielectric analysis of human stratum corneum in vivo. AB - The dielectric properties of the human skin stratum corneum (SC) in the frequency range higher than 107 Hz are not well understood because of the difficulty in selective scanning of the SC area in vivo. The present study was carried out to make clear factors responsible for the dielectric properties using a measuring system specially developed for the study of SC [S. Naito, M. Hoshi, S. Mashimo, Anal. Biochem. 251 (1997) 163-172]. We found that the dielectric properties of SC can be expressed by the linear combination of two relaxation processes and d.c. conduction. The faster relaxation is that of free water. The slower relaxation and d. c. conduction were analyzed using a model assuming interfacial polarization between dissimilar materials. We concluded that the polarization is the origin of the slower relaxation process because the experimental data could be well interpreted according to the above mechanism. We also concluded that the polarization of swelled SC locates at the interface between SC cells and the intercellular lipid layer, or at the interface between the lipophilic and the hydrophilic part of the lamellar structured intercellular lipid layer. PMID- 9729436 TI - Cloning and expression of the secA gene of a marine bacterium, Vibrio alginolyticus, and analysis of its function in Escherichia coli. AB - We report the cloning, sequencing and functional characterization of the secA gene of a marine bacterium, Vibrio alginolyticus, which has been suggested to utilize ATP and the sodium motive force for protein translocation. Oligodeoxynucleotides corresponding to highly conserved regions of Escherichia coli secA located in the high affinity ATP binding site were utilized as PCR primers to clone the secA gene of V. alginolyticus. It was shown to encode a 103.3-kDa protein. The deduced amino acid sequence of V. alginolyticus SecA (VaSecA) exhibits a high degree of identity (72.7%) to SecA of E. coli (EcSecA). The secA gene of E. coli forms an operon with upstream orfX, whereas no counterpart is present upstream of V. alginolyticus secA. Azide derepresses the EcSecA translation, whereas the level of VaSecA was unaffected by azide. Expression of VaSecA in E. coli carrying a temperature-sensitive secA mutation restored both growth and protein translocation at a non-permissive temperature. VaSecA was thus able to substitute for EcSecA despite the fact that the energy requirement for protein translocation differs between the two organisms. VaSecA was overproduced in V. alginolyticus and purified to homogeneity for N-terminal sequencing. The endogenous ATPase activity of the purified VaSecA was comparable with that of EcSecA. 1998 Elsevier Science B.V. PMID- 9729437 TI - Nonsteroidal anti-inflammatory drugs enhance glutathione S-transferase theta levels in rat colon. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) have been claimed to reduce cancer rates in oesophagus, stomach and colon of humans and laboratory animals. Recently we showed that dietary administration of NSAIDs enhanced glutathione S transferase (GST) class alpha, mu and pi levels in the upper part of the rat gastrointestinal tract, with minor effects in the colon. Enhancement of GSTs, a family of detoxification enzymes consisting of class alpha, mu, pi and theta isoforms, might be one of the mechanisms leading to cancer prevention. The recently cloned GST class theta levels have not yet been studied in this respect. We now investigated whether the NSAIDs indomethacin, relafen, sulindac, ibuprofen, piroxicam, and acetyl salicylic acid (ASA), incorporated individually into the diet at 25, 200, 320, 400, 400 and 400 mg/kg, respectively, affect gastrointestinal GSTT1-1 and GSTT2-2 levels in male Wistar rats. GSTT1-1 and GSTT2-2 levels were determined in cytosolic fractions of oesophagus, gastric, small intestinal and colonic mucosa and liver by densitometrical analyses of Western blots after immunodetection with a monoclonal (GSTT1-1) or a polyclonal (GSTT2-2) antibody. Gastric GSTT2-2 levels were induced by ibuprofen (1.6x) and indomethacin (1.5x), and colonic levels were induced by ASA (1.7x). Colonic GSTT1 1 levels were elevated by all NSAIDs tested except for relafen (1.5-6.4x). In conclusion, enhancement of colonic GSTT1-1 levels seems to be a common working mechanism of NSAIDs. Enhanced enzyme activity, which may result from these higher GSTT1-1 levels, might lead to a more efficient detoxification of potential carcinogens and hence contribute to the prevention of colon carcinogenesis. PMID- 9729438 TI - Characterization of a 76 kDa endosomal, multispanning membrane protein that is highly conserved throughout evolution. AB - We report here the identification and characterization of a human 76kDa membrane protein that is found predominantly in endosomes. This protein is related to the Saccharomyces cerevisiae EMP70 gene product, a precursor protein whose 24kDa cleavage product (p24a) was found in yeast endosome-enriched membrane fractions (Singer-Kruger et al., 1993. J. Biol. Chem. 268, 14376-14386). Northern blot analysis indicated that p76 mRNA is highly expressed in human pancreas but could be detected in all tissues examined. p76 is highly conserved throughout evolution, as related proteins have also been detected in Caenorhabditis elegans and Arabidopsis thaliana. This family of proteins has a relatively divergent, hydrophilic N-terminal domain and a well-conserved, highly hydrophobic C-terminal domain which contains nine potential membrane-spanning domains. Transiently expressed, myc-tagged human p76 appears to be localized to endosomes by virtue of its apparent colocalization with transferrin receptors and some mannose 6 phosphate receptors. Furthermore, p76 adopts a type-I topology within the membrane, with its hydrophilic N-terminus facing the lumen of cytoplasmic membranes. The structural features of p76 suggest that it may function as a channel or small molecule transporter in intracellular compartments throughout phylogeny. 1998 Elsevier Science B.V. PMID- 9729439 TI - Induction of mRNAs for glutathione synthesis-related proteins in mouse liver by low doses of gamma-rays. AB - We examined the elevation of the reduced form of glutathione (GSH)level and the induction of MRNAs for proteins involved in the synthesis and regeneration of GSH in the liver of mice after low-dose gamma-ray irradiation. The liver GSH level increased soon after irradiation with 50 cGy of gamma-rays, reached a maximum at around 12 h post-treatment. The mRNA of gamma-glutamylcysteine synthetase (gamma GCS), the rate-limiting enzyme for de novo synthesis for GSH, showed a small increase that peaked at 6 h after gamma-ray irradiation at a dose of 50 cGy. Only a small increase in gamma-GCS activity was observed throughout the 24-h post irradiation period. In the case of glutathione reductase (GR), which is involved in the regeneration of GSH from the oxidized form (GSSG), the mRNA level peaked strongly at 1 h, while the activity peaked at twice the control level 12 h after irradiation. The level of mRNA for thioredoxin (TRX), which contributes to GSH biosynthesis by supplying cysteine to the de novo pathway, peaked at 1 h and declined thereafter, while the activity peaked at 3 h and then declined sharply. These results indicate that the increase in endogenous GSH immediately following low-dose gamma-ray irradiation is predominantly due to operation of the regeneration cycle and not de novo synthesis. We also examined the dependence of mRNA induction on the gamma-ray dose. PMID- 9729440 TI - Cloning of the alphaA-crystallin genes of a blind cave form and the epigean form of Astyanax fasciatus: a comparative analysis of structure, expression and evolutionary conservation. AB - In the present study we have analyzed the integrity and expression of the alphaA crystallin gene, that codes for a major structural component of the lens, in a blind cave form of the teleostean fish, Astyanax fasciatus. This is the first alphaA-crystallin gene cloned from a teleostean fish. Sequence comparison of this cave-form gene with its epigean conspecific and with homologs of distantly related taxa has illustrated conservation of regulatory and coding regions. Although no crystallin proteins are produced in the lens of the cave form, and the mRNA of this gene could not be detected by in situ hybridization of different developmental stages, the promoter region of cave-fish alphaA-crystallin is functionally intact. The deduced amino-acid sequence of the alphaA-crystallin gene of the cave form differs from that of its epigean conspecific at only one position (139). This is within an important, small heat-shock protein-related region, HCR2. A comparison of the 5'-flanking regions of the A. fasciatus alphaA crystallin gene with the chicken homolog revealed the high conservation of lens specific regulatory sequences and further demonstrates the evolutionary conservation of this gene. 1988 Elsevier Science B.V. PMID- 9729441 TI - Impaired growth of an Escherichia coli rpe mutant lacking ribulose-5-phosphate epimerase activity. AB - We present evidence that ribulose-5-phosphate epimerase, a central metabolic enzyme acting in the non-oxidative branch of the pentose-phosphate pathway, is encoded by a gene in the dam containing operon of Escherichia coli. Enzymatic assays confirm that this gene encodes ribulose-5-phosphate epimerase activity. Disruption of the gene (rpe) causes loss of enzymatic activity and renders the rpe mutant unable to utilize single pentose sugars, indicating that rpe supplies the only ribulose-5-phosphate epimerase activity in E. coli. Growth of the rpe mutant is impaired in complex LB medium and severely impaired in minimal medium containing glycolytic carbon sources or gluconate. Enrichment with casamino acids abolishes or strongly relieves growth suppression in minimal medium. Aspartate counteracts the impaired growth in glycolytic carbon sources but not in gluconate. We suggest that the absence of the Rpe enzyme causes changes in the pentose-phosphate levels which alter the regulation of (a) metabolic enzyme(s) and thereby cause growth suppression and that the severity of growth suppression is related to the internal concentration of pentose-phosphates. Target enzymes for negative regulation may be located in the early parts of the Embden-Meyerhof Parnas pathway and of the Entner-Doudoroff pathway and/or of carbohydrate transport systems feeding sugars into these sections of central metabolic pathways. PMID- 9729442 TI - Identification and cloning of a developmentally regulated Cryptosporidium parvum gene by differential mRNA display PCR. AB - To identify Cryptosporidium parvum genes expressed during intracellular development, differential mRNA display was used to detect differences in gene expression between mock-infected and C. parvum-infected human epithelial cells. A reproducible band present only in C. parvum-infected cells, ddHC-23, was isolated and cloned. Southern blot analysis demonstrated that ddHC-23 represented a C. parvum gene. RT-PCR revealed that HC-23 mRNA levels decreased from 6 to 12h post infection (pi), were maximally expressed at 24h pi, and returned to low levels at 48 and 72h pi. Northern blot analysis determined that the approx. 3.6kb transcript is expressed by sporozoites prior to invasion of epithelial cells. Screening of a C. parvum genomic library with ddHC-23 isolated a genomic subclone which contained a 2790bp ORF, uninterrupted by introns. Sequence analysis indicated that the encoded protein, which displayed no similarity to any sequences in the public databases, contained a high proportion of polar amino acids, with the most abundant being Asp (17.3%), Ser (15.8%) and Gly (8.1%). Numerous potential sites for posttranslational modification were present including: casein kinase II and protein kinase C phosphorylation sites, N myristolation sites and N-glycosylation sites. These findings demonstrate the usefulness of differential mRNA display for identifying developmentally regulated C. parvum genes within the background of genes expressed by the host cell. 1998 Elsevier Science B.V. PMID- 9729443 TI - Expression of grass carp growth hormone by baculovirus in silkworm larvae. AB - A total of five recombinant Bombyx mori nuclear polyhedrosis viruses (BMNPV) carrying the grass carp (Ctenopharyngodon idellus) growth hormone (GH) cDNA were constructed in this study. Two of them were able to express the hormone up to a level of 12 microgram/ml medium when cultured B. mori cells were infected for 4 days. Inoculation of the viruses into silkworm (B. mori) host significantly increased the level of GH achievable. The amount of hormone produced per larva was estimated to be around 1 mg. The recombinant grass carp GH had immunological and biological activities similar to the native hormone. The N-terminal sequence of the recombinant hormone was the same as the native one, indicating that the fish signal peptide was correctly processed by the insect cells. Silkworm powder prepared from larvae infected with the recombinant virus was used as food supplement for fish. Compared with the control, this dietary supplement was effective in increasing the growth rate of juvenile carp. PMID- 9729444 TI - Factors that cause the beta-anomeric preference of Na+/glucose cotransporter for intestinal transport of monosaccharide conjugates. AB - The intestinal transport of glucose- and galactose-conjugated acetaminophen (APAP glycoside) by Na+/glucose cotransporter (SGLT1) was studied. SGLT1-mediated transport of APAP glycosides preferred glucoside>galactoside and beta anomer>alpha-anomer. These preferences agree with previous studies. NMR spectroscopic and molecular modeling studies indicated that the conformation of the glucose ring of alpha- and beta-glucosides of APAP, as well as glycosides in previous studies, is in the 4C1 chair form, the same form as glucose itself. Molecular dynamics analysis also indicated that the glucose ring was in the 4C1 chair form, and that there are differences between the rotational spaces of aglycones and hydroxy groups of glucose moieties between anomers. Therefore, we conclude that the beta-anomeric preference of glucose conjugate transport by SGLT1 is not due to the conformation of the glucose ring, but to the configuration of the aglycone at C-1 of the monosaccharide moiety. PMID- 9729445 TI - The nirQ gene, which is required for denitrification of Pseudomonas aeruginosa, can activate the RubisCO from Pseudomonas hydrogenothermophila. AB - Two putative ATP-binding proteins encoded in the gene cluster for the Calvin cycle of Pseudomonas hydrogenothermophila (cbbQ) and for the denitrification of Pseudomonas aeruginosa (nirQ) have been found to be similar. The cbbQ gene has been shown to activate the RubisCO from P. hydrogenothermophila in E. coli. The nirQ was functionally substituted for cbbQ. The nirQ gene restored the anaerobic growth and the NOR activity of the nirQOP mutant of P. aeruginosa, while the cbbQ gene did not. PMID- 9729447 TI - Corrigendum to 'The new EPR molecular oxygen probe fusinite is not toxic to cells'. PMID- 9729446 TI - Effect of pyridoxine and insulin administration on brain glutamate dehydrogenase activity and blood glucose control in streptozotocin-induced diabetic rats. AB - Blood glucose level and kinetic parameters of glutamate dehydrogenase (GDH) were measured in the cerebellum, brain stem and cerebral cortex of control, insulin treated, pyridoxine treated, pyridoxine and insulin treated and untreated streptozotocin-diabetic rats. The combined administration of insulin and pyridoxine was found to be better in controlling the hyperglycaemia. Insulin with pyridoxine treatment brought back the increased maximal velocity of GDH during diabetes to control state. Also, there was an increase in Michaelis-Menten constant. These results suggest that pyridoxine and insulin together serve a better control for diabetes. PMID- 9729449 TI - Management of infections due to the varicella-zoster virus. 11th consensus conference on anti-infectious therapy of the French-speaking Society of Infectious Diseases (SPILF). PMID- 9729450 TI - Working memory deficits of reading disabled children. AB - Aims of the study were to investigate the specificity of reading disabled childrens deficits in working memory capacity and to pursue whether their deficits could be accounted for by deficient processing or impairments in verbal short-term storage capacity. A group of 10-year-old reading disabled children was compared with two groups of normal reading children, matched for chronological age and reading age, respectively. Measures for working memory capacity, short term capacity and processing speed related to the language and to the numerical domain were administered. Results indicated that reading disabled children performed worse on all measures of working memory capacity, irrespective of the domain which these measures reflected. Their poorer performance could neither be explained by inefficient processing nor to their deficits in verbal short-term storage capacity. Reading disabled children seem to have a general lack of capacity for the concurrent processing and storage of verbal information. PMID- 9729451 TI - What's in a name: Children's knowledge about the letters in their own names. AB - Two studies were performed to determine whether children's experiences with their own names boost their knowledge about the components of the name, the letters. The children in Study One showed a significant superiority for the initial letter of their own first name in tests of letter-name, but not letter-sound, knowledge. This pattern was found for Australian first graders (mean age 5 years, 5 months), U.S. kindergartners (mean age 5 years, 8 months), and U.S. preschoolers (mean age 4 years, 10 months). Study Two, with U.S. preschoolers (mean age 4 years, 11 months), again revealed an advantage for the initial letter of a child's first name in knowledge of letter names but not knowledge of letter sounds. Moreover, the children were better at printing the initial letter of their own first name than other letters. The results show that different factors are involved in the learning of letter names and letter sounds. They further suggest that children use letter-based strategies with their own names at a time when they are often considered to be "logographic" readers. PMID- 9729452 TI - Bidirectional relations of phonological sensitivity and prereading abilities: evidence from a preschool sample. AB - Children's phonological sensitivity is a strong predictor of the development of reading skills. Recent evidence indicates that phonological sensitivity and reading are reciprocally related. That is, phonological sensitivity facilitates the development of early reading and early reading facilitates the development of phonological sensitivity. Whereas evidence for this reciprocal relation has come from studies with school-age children, this study examined the relation between phonological sensitivity and letter knowledge in 97 middle-income 4- and 5-year old children in a 1-year longitudinal study. Multiple regression analyses revealed that phonological sensitivity predicted growth in letter knowledge, and letter knowledge predicted growth in phonological sensitivity when controlling for children's age and oral language abilities. These results indicate that the reciprocal relation between reading and phonological sensitivity is present relatively early in the development of literacy skills, prior to the onset of formal reading instruction. PMID- 9729454 TI - Kinetics of elementary Ca2+ puffs evoked in Xenopus oocytes by different Ins(1,4,5)P3 receptor agonists. AB - Elementary Ca2+ puffs form the basic building blocks of global Ins(1, 4,5)P3 evoked Ca2+ signals. In Xenopus oocytes, Ca2+ puffs evoked by the high-affinity agonist adenophostin were shorter and smaller than puffs evoked by Ins(1,4,5)P3 and the lower affinity analogue Ins(2,4, 5)P3. Agonist-specific mechanisms, therefore, play a role in shaping local Ca2+ release events, but termination of Ca2+ flux is not delimited simply by agonist dissociation. PMID- 9729455 TI - Channelling of intermediates in the biosynthesis of phosphatidylcholine and phosphatidylethanolamine in mammalian cells. AB - Previous studies with electropermeabilized cells have suggested the occurrence of metabolic compartmentation and Ca2+-dependent channeling of intermediates of phosphatidylcholine (PC) biosynthesis in C6 rat glioma cells. With a more accessible permeabilization technique, we investigated whether this is a more general phenomenon also occurring in other cell types and whether channeling is involved in phosphatidylethanolamine (PE) synthesis as well. C6 rat glioma cells, C3H10T12 fibroblasts and rat hepatocytes were permeabilized with Staphylococcus aureus alpha-toxin, and the incorporation of the radiolabelled precursors choline, phosphocholine (P-choline), ethanolamine and phosphoethanolamine (P-EA) into PC and PE were measured both at high and low Ca2+ concentrations. In glioma cells, permeabilization at high Ca2+ concentration did not affect [14C]choline or [14C]P-choline incorporation into PC. However, reduction of free Ca2+ in the medium from 1.8 mM to <1 nM resulted in a dramatic increase in [14C]P-choline incorporation into permeabilized cells, whereas [14C]choline incorporation remained unaffected. Also, in fibroblasts, reduction of extracellular Ca2+ increased [14C]P-choline and [14C]P-EA incorporation into PC and PE respectively. In hepatocytes, a combination of alpha-toxin and low Ca2+ concentration severely impaired [14C]choline incorporation into PC. Therefore, alpha-toxin-permeabilized hepatocytes are not a good model in which to study channeling of intermediates in PC biosynthesis. In conclusion, our results indicate that channeling is involved in PC synthesis in glioma cells and fibroblasts. PE synthesis in fibroblasts is also at least partly dependent on channeling. PMID- 9729456 TI - Restricting mobility of Gsalpha relative to the beta2-adrenoceptor enhances adenylate cyclase activity by reducing Gsalpha GTPase activity. AB - The beta2-adrenoceptor (beta2AR) activates the G-protein Gsalpha to stimulate adenylate cyclase (AC). Fusion of the beta2AR C-terminus to the N-terminus of Gsalpha (producing beta2ARGsalpha) markedly increases the efficiency of receptor/G-protein coupling compared with the non-fused state. This increase in coupling efficiency can be attributed to the physical proximity of receptor and G protein. To determine the optimal length for the tether between receptor and G protein we constructed fusion proteins from which 26 [beta2AR(Delta26)Gsalpha] or 70 [beta2AR(Delta70)Gsalpha] residues of the beta2AR C-terminus had been deleted and compared the properties of these fusion proteins with the previously described beta2ARGsalpha. Compared with beta2ARGsalpha, basal and agonist stimulated GTP hydrolysis was markedly decreased in beta2AR(Delta70)Gsalpha, whereas the effect of the deletion on binding of guanosine 5'-[gamma thio]triphosphate (GTP[S]) was relatively small. Surprisingly, deletions did not alter the efficiency of coupling of the beta2AR to Gsalpha as assessed by GTP[S] sensitive high-affinity agonist binding. Moreover, basal and ligand-regulated AC activities in membranes expressing beta2AR(Delta70)Gsalpha and beta2AR(Delta26)Gsalpha were higher than in membranes expressing beta2ARGsalpha. These findings suggest that restricting the mobility of Gsalpha relative to the beta2AR results in a decrease in G-protein inactivation by GTP hydrolysis and thereby enhanced activation of AC. PMID- 9729457 TI - Increased choline transport in erythrocytes from mice infected with the malaria parasite Plasmodium vinckei vinckei. AB - Parasitized erythrocytes from mice infected with the murine malaria parasite Plasmodium vinckei vinckei showed a marked increase in the rate of influx of choline compared with erythrocytes from uninfected mice. In contrast, uninfected erythrocytes from P. vinckei-infected animals transported choline at the same rate as those from uninfected mice. The increased influx of choline into parasitized cells was via two discrete routes. One was a saturable pathway with a Km similar to that of the choline carrier of normal erythrocytes but a Vmax approx. 20-fold higher than that observed in uninfected cells. The other was a non-saturable pathway inhibited by furosemide. At choline concentrations within the normal physiological plasma concentration range, the former pathway contributed approx. two-thirds and the latter approx. one-third of the influx of choline into parasitized cells. The characteristics of the furosemide-sensitive pathway were similar to those of a broad-specificity pathway that is induced in human erythrocytes infected in vitro with Plasmodium falciparum. The results of this study rule out the possibility that the induced transport pathway of P. falciparum-infected erythrocytes is an artifact arising in vitro from the long term culture of parasitized cells and provide evidence that this pathway makes a significant contribution to the uptake of choline into the parasitized cells of malaria-infected animals. PMID- 9729453 TI - Regulation of serum amyloid A protein expression during the acute-phase response. AB - The acute-phase (AP) serum amyloid A proteins (A-SAA) are multifunctional apolipoproteins which are involved in cholesterol transport and metabolism, and in modulating numerous immunological responses during inflammation and the AP response to infection, trauma or stress. During the AP response the hepatic biosynthesis of A-SAA is up-regulated by pro-inflammatory cytokines, and circulating concentrations can increase by up to 1000-fold. Chronically elevated A-SAA concentrations are a prerequisite for the pathogenesis of secondary amyloidosis, a progressive and fatal disease characterized by the deposition in major organs of insoluble plaques composed principally of proteolytically cleaved A-SAA, and may also contribute to physiological processes that lead to atherosclerosis. There is therefore a requirement for both positive and negative control mechanisms that permit the rapid induction of A-SAA expression until it has fulfilled its host-protective function(s) and subsequently ensure that its expression can be rapidly returned to baseline. These mechanisms include modulation of promoter activity involving, for example, the inducer nuclear factor kappaB (NF-kappaB) and its inhibitor IkappaB, up-regulatory transcription factors of the nuclear factor for interleukin-6 (NF-IL6) family and transcriptional repressors such as yin and yang 1 (YY1). Post-transcriptional modulation involving changes in mRNA stability and translation efficiency permit further up- and down-regulatory control of A-SAA protein synthesis to be achieved. In the later stages of the AP response, A-SAA expression is effectively down-regulated via the increased production of cytokine antagonists such as the interleukin-1 receptor antagonist (IL-1Ra) and of soluble cytokine receptors, resulting in less signal transduction driven by pro-inflammatory cytokines. PMID- 9729458 TI - Differences in the autocatalytic cleavage of pro-PC2 and pro-PC3 can be attributed to sequences within the propeptide and Asp310 of pro-PC2. AB - PC2 and PC3 are subtilisin-like proteases involved in the maturation of prohormones and proneuropeptides within neuroendocrine cells. They are synthesized as zymogens that undergo autocatalytic maturation within the secretory pathway. Maturation of pro-PC2 is slow (t12 >8 h), exhibits a pH optimum of 5.5 and is dependent on calcium (K0.5 2 mM), while pro-PC3 maturation is relatively rapid (t12 15 min), exhibits a neutral pH optimum and is not calcium dependent. These differences in the rates and optimal conditions for activation of the proteases may contribute to the diversity of products generated by these proteases in different cell types. Although highly similar, there are two major differences between pro-PC2 and pro-PC3: the presence of an aspartate at position 310 in pro-PC2 compared with asparagine at the equivalent position in pro-PC3 (and all other members of the subtilisin family), and the N-terminal propeptides, which exhibit low sequence identity (30%). With a view to establishing the structural features that might be responsible for these differences in the maturation of pro-PC2 and pro-PC3, Asp310 in pro-PC2 was mutated to Asn, and Asn309 in pro-PC3 was mutated to Asp. Chimaeric proteins were also made consisting of the pro-region of PC2 fused to the mature portion of PC3 and the pro-region of PC3 fused to the mature region of PC2. The wild-type and mutant DNA constructs were then transcribed and translated in an in vitro system capable of supporting maturation of pro-PC2 and pro-PC3. The results demonstrated that Asp310 of pro-PC2 is responsible for the acidic pH optimum for maturation. Thus changing Asp310 to Asn shifted the pH optimum for maturation to pH 7.0. However, changing Asn309 of pro-PC3 to Asp had no effect on the optimum pH for maturation of pro-PC3. A chimaeric construct containing the propeptide of pro-PC2 attached to PC3 shifted the pH optimum for maturation from pH 7.0 to 6.0 and slowed down the rate of maturation (t12 >8 h). When attached to PC2, the pro region of pro-PC3 had no effect on the optimum pH for maturation (pH 5.5-6.0), but it did accelerate the rate of maturation (t12 2 h). These results demonstrate that Asp310 and the pro-region of pro-PC2 contribute to the acidic pH optimum and low rate of maturation of this zymogen relative to its closely related homologue PC3. PMID- 9729459 TI - Species barrier in prion diseases: a kinetic interpretation based on the conformational adaptation of the prion protein. AB - Prion diseases are thought to result from the conformational change of the normal cellular prion protein to a pathogenic protease-resistant isoform. However, brain extracts not containing the protease-resistant isoform of the prion protein can be infectious following interspecies transmission. The 'protein-only' hypothesis of pathogenesis is extended to provide possible explanations which could be interpreted in terms of a different infectious agent. It is proposed that normal cellular protein (PrPC) may be transformed into a form (PrP*) that is conformationally distinct from the host-specific abnormal isoform (PrPSc). In infection from a heterologous donor, the dimeric forms of heterologous PrPSc, which may catalyse the formation of host PrP* from PrPC, host PrP* and host PrPSc are all considered to be capable of catalysing, to some extent, the conversion of PrPC into PrPSc. However, depending on the species involved, PrP* may, or may not, be pathogenic, and may, or may not, be sensitive to proteolysis. It is shown, by numerical integration of the differential rate equations derived from this model, that a strain may be stabilized after two or three passages through a different species and that transmission might occur in the absence of detectable protease-resistant prion protein. The natural transmission of scrapie to cattle is discussed in relation to the model. PMID- 9729460 TI - Specific alterations in levels of mannose 6-phosphorylated glycoproteins in different neuronal ceroid lipofuscinoses. AB - Mannose 6-phosphate (Man-6-P) is a carbohydrate modification that is generated on newly synthesized lysosomal proteins. This modification is specifically recognized by two Man-6-P receptors that direct the vesicular transport of the lysosomal enzymes from the Golgi to a prelysosomal compartment. The Man-6-P is rapidly removed in the lysosome of most cell types; however, in neurons the Man-6 P modification persists. In this study we have examined the spectrum of Man-6-P containing glycoproteins in brain specimens from patients with different neuronal ceroid lipofuscinoses (NCLs), which are progressive neurodegenerative disorders with established links to defects in lysosomal catabolism. We find characteristic alterations in the Man-6-P glycoproteins in specimens from late-infantile (LINCL), juvenile (JNCL) and adult (ANCL) patients. Man-6-P glycoproteins in LINCL patients were similar to controls, with the exception that the band corresponding to CLN2, a recently identified lysosomal enzyme whose deficiency results in this disease, was absent. In an ANCL patient, two Man-6-P glycoproteins were elevated in comparison with normal controls, suggesting that this disease also results from a perturbation in lysosomal hydrolysis. In JNCL, total levels of Man-6-P glycoproteins were 7-fold those of controls. In general this was reflected by increased lysosomal enzyme activities in JNCL but three Man 6-P glycoproteins were elevated to an even greater degree. These are CLN2 and the unidentified proteins that are also highly elevated in the ANCL. PMID- 9729461 TI - Mutation of tyrosine-194 and lysine-198 in the catalytic site of pig 3alpha/beta,20beta-hydroxysteroid dehydrogenase. AB - Pig 3alpha/beta,20beta-hydroxysteroid dehydrogenase is an NADPH-dependent enzyme that catalyses the reduction of ketones on steroids and aldehydes and ketones on various xenobiotics, like its homologue carbonyl reductase. 3alpha/beta,20beta Hydroxysteroid dehydrogenase and carbonyl reductase are members of the short chain dehydrogenases/reductase family, in which a tyrosine residue and a lysine residue have been identified as catalytically important. In pig 20beta hydroxysteroid dehydrogenase these residues are tyrosine-194 and lysine-198. Here we report the effect on the reduction of two ketone and two aldehyde substrates by pig 3alpha/beta,20beta-hydroxysteroid dehydrogenase in which tyrosine-194 has been mutated to phenylalanine and cysteine, and lysine-198 has been mutated to isoleucine and arginine. Mutants with phenylalanine-194 or isoleucine-198 are inactive. Depending on the substrate, the mutant with cysteine-194 has a catalytic efficiency of 0.4-1% and the mutant with arginine-198 has a catalytic efficiency of 4-23% of the wild-type enzyme. We also mutated tyrosine-81 and tyrosine-253 to phenylalanine. Although both tyrosines are conserved in 3alpha/beta,20beta-hydroxysteroid dehydrogenase and carbonyl reductase, depending on the substrate, the mutant enzymes are as active as, or more active than, wild type enzyme. PMID- 9729462 TI - Muscle-specific mRNA isoform encodes a protein composed mainly of the N-terminal 175 residues of type 2 Ins(1,4,5)P3 receptor. AB - We have found a novel isoform of the mouse type 2 Ins(1,4,5)P3 receptor [Ins(1,4,5)P3R] mRNA by reverse transcriptase-mediated PCR analysis. The novel isoform, which was expressed specifically in skeletal muscle and heart, was generated by the inclusion of a novel exon. As this exon contains a stop codon, the isoform encodes a putative protein (designated TIPR) consisting of 175 acid residues of the type 2 Ins(1,4,5)P3R and the following six residues derived from this exon. We transfected the cDNA of this isoform into COS-7 cells; these cells expressed a 24 kDa protein that was recognized by an antibody against TIPR produced in Escherichia coli. The isoform encoding TIPR was also found in human skeletal muscle and heart. The N-terminal region of Ins(1,4,5)P3R is suggested to have a role in ligand binding and to interact with the C-terminal channel domain of Ins(1,4,5)P3R itself. TIPR might regulate the Ins(1,4,5)P3 signal pathway in both muscles. PMID- 9729463 TI - Hepatocyte nuclear factor 6: organization and chromosomal assignment of the rat gene and characterization of its promoter. AB - Hepatocyte nuclear factor 6 (HNF-6) is the prototype of a family of tissue specific transcription factors characterized by a bipartite DNA-binding domain consisting of a single cut domain and a novel type of homeodomain. We have previously cloned rat cDNA species coding for two isoforms, HNF-6alpha (465 residues) and beta (491 residues), which differ only by the length of the spacer between the two DNA-binding domains. We have now localized the rat Hnf6 gene to chromosome 8q24-q31 by Southern blotting of DNA from somatic cell hybrids and by fluorescence in situ hybridization. Cloning and sequencing of the rat gene showed that the two HNF-6 isoforms are generated by alternative splicing of three exons that are more than 10 kb apart from each other. Exon 1 codes for the N-terminal part and the cut domain, exon 2 codes for the 26 HNF-6beta-specific amino acids, and exon 3 codes for the homeodomain and the C-terminal amino acids. The transcription initiation site was mapped by ribonuclease protection and 5' rapid amplification of cDNA ends. Transfection experiments showed that promoter activity was contained within 0.75 kb upstream of the transcription initiation site. This activity was detected by the transfection of liver-derived HepG2 cells, but not of Rat-1 fibroblasts, suggesting that the promoter is sufficient to confer liver-specific expression. PMID- 9729464 TI - Plant mitochondrial pyruvate dehydrogenase complex: purification and identification of catalytic components in potato. AB - The pyruvate dehydrogenase complex (mPDC) from potato (Solanum tuberosum cv. Romano) tuber mitochondria was purified 40-fold to a specific activity of 5.60 micromol/min per mg of protein. The activity of the complex depended on pyruvate, divalent cations, NAD+ and CoA and was competitively inhibited by both NADH and acetyl-CoA. SDS/PAGE revealed the complex consisted of seven polypeptide bands with apparent molecular masses of 78, 60, 58, 55, 43, 41 and 37 kDa. N-terminal sequencing revealed that the 78 kDa protein was dihydrolipoamide transacetylase (E2), the 58 kDa protein was dihydrolipoamide dehydrogenase (E3), the 43 and 41 kDa proteins were alpha subunits of pyruvate dehydrogenase, and the 37 kDa protein was the beta subunit of pyruvate dehydrogenase. N-terminal sequencing of the 55 kDa protein band yielded two protein sequences: one was another E3; the other was similar to the sequence of E2 from plant and yeast sources but was distinctly different from the sequence of the 78 kDa protein. Incubation of the mPDC with [2-14C]pyruvate resulted in the acetylation of both the 78 and 55 kDa proteins. PMID- 9729465 TI - An array of binding sites for hepatocyte nuclear factor 4 of high and low affinities modulates the liver-specific enhancer for the human alpha1 microglobulin/bikunin precursor. AB - Alpha1-Microglobulin and bikunin are two plasma glycoproteins encoded by a gene for alpha1-microglobulin/bikunin precursor (AMBP). The strict liver-specific transcription of the AMBP gene is controlled by an elaborate and remote enhancer made of six clustered boxes numbered 1 to 6 (core enhancer) that are binding sites for the hepatocyte-enriched nuclear factors HNF-1, HNF-4, HNF-3, HNF-1, HNF 3 and HNF-4 respectively. Three further boxes, 7 to 9, have now been found in the enhancer area in a position 5' of box 2, 5' of box 1 and 3' of box 6, respectively. Electrophoretic mobility-shift assays with nuclear extracts from the HepG2 hepatoma cell line demonstrated that boxes 7 and 8 are both functional HNF-4-binding sites of high and low affinity respectively, whereas no binding capacity of box 9 was detected by this method. Transfection of HepG2 and Chinese hamster ovary cells with chloramphenicol acetyltransferase constructs harbouring the core or extended AMBP enhancer with wild-type or mutated boxes and co transfection with expression plasmids for a wild-type or defective HNF-4 identified box 7 as an essential element for the basal activity of this enhancer. The response of boxes 7 and 8 varies with the level of HNF-4 in cells. Box 9 exhibits a repressor activity that can be detected when box 8 is ablated. In vivo this corresponds to conditions of low box 8 occupancy when the intracellular level of HNF-4 is limited. These results reinforce the view that the AMBP enhancer is a quite elaborate and unusual example of a modular enhancer whose activity is fine-tuned by the level of cognate nuclear factors in the cell. PMID- 9729466 TI - Structural determination of a 5-O-methyl-deaminated neuraminic acid (Kdn) containing polysaccharide isolated from Sinorhizobium fredii. AB - The structure of a polysaccharide from Sinorhizobium fredii SVQ293, a thiamine auxotrophic mutant of S. fredii HH103, has been determined. This polysaccharide was isolated following the protocol for lipopolysaccharide extraction. On the basis of monosaccharide analysis, methylation analysis, fast atom bombardment MS, collision-induced dissociation tandem MS, one-dimensional 1H and 13C NMR and two dimensional NMR experiments, the structure was shown to consist of the following trisaccharide repeating unit-->2)-alpha-d-Galp-(1-->2)-beta-d-Ribf-(1-->9)-alpha 5-O-Me-++ +Kdnp- (2-->, in which Kdn stands for deaminated neuraminic acid; 25% of the Kdn residues are not methylated. The structure of this polysaccharide is novel and this is the first report of the presence of Kdn in a rhizobial polysaccharide, as well as being the first structure described containing 5-O-Me Kdn. This Kdn-containing polysaccharide is not present in the wild-type strain HH103, which produces a 3-deoxy-d-manno-2-octulosonic acid (Kdo)-rich polysaccharide. We conclude that it is likely that the appearance of this new Kdn containing polysaccharide is a consequence of the mutation. PMID- 9729468 TI - Nucleotide binding by the nitrogenase Fe protein: a 31P NMR study of ADP and ATP interactions with the Fe protein of Klebsiella pneumoniae. AB - Investigation of the interaction of MgADP- and MgATP2- with the Fe protein of Klebsiella pneumoniae nitrogenase by 31P NMR showed that the adenine nucleotides are reversibly bound in slow exchange with free nucleotides. Dissociation of the MgADP--Fe protein complex was slow enough to enable its isolation by gel filtration, thus permitting the assignment of resonances to bound nucleotides. Spectra of ADP bound to Kp2 were similar to spectra of ADP bound to the myosin motor domain. Oxidative inactivation of a Kp2-MgADP- complex with excess ferricyanide ion eliminated exchange between bound and free ADP, indicating that the intact iron sulphur cluster, located 20 A from the binding sites, is required for the reversible binding of MgADP-. A change in conformation on controlled oxidation of Kp2 with indigocarmine increased the chemical shift of the beta phosphate resonance of bound MgADP-. Both oxidized and reduced conformers were observed transiently in the absence of dithionite. The 31P resonances of both the beta and gamma phosphates of bound MgATP2- indicated major changes in environment and labilization of both groups on binding to the Fe protein. Highly purified Kp2 slowly hydrolysed ATP, resulting in mixtures of bound nucleotides. Partial occupation of Kp2 MgATP2--binding sites (N=1.9+/-0.2, Kd=145 microM) in concentrated protein solutions was demonstrated by flow dialysis. Scatchard plots of data for bound and free ligand obtained after equilibration with Kp2 were linear and no co-operative interactions were detected. We conclude that MgADP- stabilizes the oxidized Fe protein conformer and this conformation in turn triggers the dissociation of the Fe protein from the MoFe protein in the rate limiting step of the overall process of dinitrogen reduction. PMID- 9729467 TI - Insulin stimulates the tyrosine dephosphorylation of docking protein p130cas (Crk associated substrate), promoting the switch of the adaptor protein crk from p130cas to newly phosphorylated insulin receptor substrate-1. AB - The docking protein p130(cas) (Crk-associated substrate) forms a stable complex with the adaptor protein CrkII in a tyrosine-phosphorylation-dependent manner. Insulin-induced tyrosine phosphorylation of insulin receptor substrates results in the redistribution of CrkII between p130(cas) and insulin receptor substrate 1. A decrease in the association between CrkII and p130(cas) in response to insulin stimulation was detected in CHO cells stably expressing insulin receptor or insulin receptor substrate-1, and in L6 rat myoblasts. Along with the decrease in the association of CrkII with p130(cas), the amount of tyrosine-phosphorylated insulin receptor substrate-1 co-precipitated with CrkII increased in all cell types studied. The insulin-induced decrease in the CrkII-p130(cas) association was further confirmed by Far Western Blot analysis with the Src homology 2 (SH2) domain of CrkII. Insulin regulates the association of CrkII with p130(cas) by tyrosine dephosphorylation of p130(cas) and co-ordinated tyrosine phosphorylation of insulin receptor substrate-1. Tyrosine-phosphorylated insulin receptor substrate-1 serves as a docking protein for multiple adaptor proteins and competes with p130(cas) for CrkII. PMID- 9729470 TI - Structure and organization of the human theta-class glutathione S-transferase and D-dopachrome tautomerase gene complex. AB - The structure and organization of the human Theta-class glutathione S-transferase (GST) genes have been determined. GSTT1 and GSTT2 are separated by approx. 50 kb. They have a similar structure, being composed of five exons with identical exon/intron boundaries. GSTT1 is 8.1 kb in length, while GSTT2 is only 3.7 kb. The GSTT2 gene lies head-to-head with a gene encoding d-dopachrome tautomerase (DDCT), which extends over 8.5 kb and contains four exons. The sequence between GSTT2 and DDCT may contain a bidirectional promoter. The GSTT2 and DDCT genes have been duplicated in an inverted repeat. Sequence analysis of the duplicated GSTT2 gene has identified an exon 2/intron 2 splice site abnormality and a premature translation stop signal at codon 196. These changes suggest that the duplicate gene is a pseudogene, and it has been named GSTT2P. PMID- 9729469 TI - Human and mouse Gpi1p homologues restore glycosylphosphatidylinositol membrane anchor biosynthesis in yeast mutants. AB - Glycosylphosphatidylinositol (GPI) represents an important anchoring molecule for cell surface proteins. The first step in its synthesis is the transfer of N acetylglucosamine (GlcNAc) from UDP to phosphatidylinositol (PI). The products of three mammalian genes, PIG-A, PIG-C and PIG-H, have previously been shown to be involved in the putative enzymic complex. Here we report the cloning of human and mouse cDNAs encoding a fourth participant in the GlcNAc transfer reaction which are homologues of the Saccharomyces cerevisiae and Schizosaccharomyces pombe Gpi1 proteins. To provide evidence for their function, these proteins were expressed in GPI1-disrupted yeast strains. In Sacch. cerevisiae, where GPI1 disruption results in a temperature-sensitive phenotype and abolishes in vitro GlcNAc-PI synthesis, restoration of growth could be demonstrated in a temperature-dependent manner. In addition, in vitro GlcNAc-PI synthetic activity was again detectable. In Schiz. pombe, gpi1+ disruption is lethal. Using random spore analysis, we were able to show that the mammalian GPI1 homologues can rescue haploids harbouring the lethal gpi1+::his7+ allele. Our data demonstrate that the genes identified are indeed involved in the first step of GPI biosynthesis, and allow conclusions about a specific function for Gpi1p in stabilizing the enzymic complex. The finding that, despite a low degree of identity, the mammalian Gpi1 proteins are able to participate in the yeast GlcNAc-PI synthetic machinery as heterologous components further demonstrates that GPI biosynthesis has been highly conserved throughout evolution. PMID- 9729471 TI - Tyrosine-phosphorylation-dependent and rho-protein-mediated control of cellular phosphatidylinositol 4,5-bisphosphate levels. AB - The polyphosphoinositide PtdIns(4,5)P2, best known as a substrate for phospholipase C isozymes, has recently been recognized to be involved in a variety of other cellular processes. The aim of this study was to examine whether the cellular levels of this versatile phospholipid are controlled by tyrosine phosphorylation. The studies were performed in human embryonic kidney (HEK)-293 cells stably expressing the M3 muscarinic acetylcholine receptor. Inhibition of tyrosine phosphatases by pervanadate induced an up-to-approx.-2. 5-fold increase in the total cellular level of PtdIns(4,5)P2, which was both time- and concentration-dependent. In contrast, the tyrosine kinase inhibitors, genistein and tyrphostin 23, caused a rapid and specific fall in the cellular PtdIns(4,5)P2 level and prevented the stimulatory effect of pervanadate on PtdIns(4,5)P2 formation. Inactivation of Rho proteins by Clostridium difficile toxin B caused a similar fall in the HEK-293 cell PtdIns(4,5)P2 level, which was not altered by additional genistein treatment. Furthermore, toxin B treatment abolished the pervanadate-induced increase in PtdIns(4,5)P2 levels. As PtdIns(4,5)P2 is an essential stimulatory cofactor for phospholipase D (PLD) enzymes, we finally examined the effects of the agents regulating PtdIns(4,5)P2 levels on PLD activity in HEK-293 cells. Inhibition of tyrosine phosphatases by pervanadate caused an increase in PLD activity, which was susceptible to genistein and tyrphostin 23, and which was abolished by prior treatment with toxin B. In conclusion, the data presented indicate that the cellular level of the multifunctional phospholipid, PtdIns(4,5)P2, in HEK-293 cells is controlled by a tyrosine-kinase-dependent mechanism and that this process apparently involves Rho proteins, as found similarly for tyrosine-phosphorylation-induced PLD activation. PMID- 9729472 TI - Thyroid hormone regulation of the Na+/glucose cotransporter SGLT1 in Caco-2 cells. AB - The expression of the Na+/glucose cotransporter (SGLT1) in response to thyroid hormone [3,5,3'-tri-iodo-l-thyronine (T3)] was investigated in the enterocytic model cell line Caco-2/TC7. In differentiated cells, T3 treatment induces an average 10-fold increase in glucose consumption as well as a T3 dose-dependent increase in SGLT1 mRNA abundance. Only cells grown on glucose-containing media, but not on the non-metabolizable glucose analogue alpha-methylglucose (AMG), could respond to T3-treatment. The Vmax parameter of AMG transport was enhanced 6 fold by T3 treatment, whereas the protein abundance of SGLT1 was unchanged. The role of Na+ recycling in the T3-related activation of SGLT1 activity was suggested by both the large increase in Na+/K+ATPase protein abundance and the inhibition, down to control levels, of AMG uptake in ouabain-treated cells. Further investigations aimed at identifying the presence of a second cotransporter that could be expressed erroneously in the colon cancer cell line were unsuccessful: T3-treatment did not modify the sugar-specificity profile of AMG transport and did not induce the expression of SGLT2 as assessed by reverse transcription-PCR. Our results show that T3 can stimulate the SGLT1 cotransport activity in Caco-2 cells. Both transcriptional and translational levels of regulation are involved. Finally, glucose metabolism is required for SGLT1 expression, a result that contrasts with the in vivo situation and may be related to the fetal phenotype of the cells. PMID- 9729474 TI - Neuropeptide regulation of biosynthesis of the juvenoid, methyl farnesoate, in the edible crab, Cancer pagurus. AB - The neuropeptide mandibular organ (MO)-inhibiting hormone (MO-IH), synthesized and secreted from the X-organ-sinus-gland complex of the eyestalk, regulates the biosynthesis of the putative crustacean juvenile hormone, methyl farnesoate (MF). Using radiolabelled acetate as a precursor for isoprenoid biosynthesis, farnesoic acid (FA), farnesol, farnesal, MF and geranyl geraniol were detected in MOs cultured for 24 h. Treatment of MOs with extract of sinus gland inhibited the final step of biosynthesis of MF, catalysed by FA O-methyltransferase. Additionally, treatment of MOs with purified MO-IH exhibited a dose-dependent inhibition of this final step of MF synthesis. The extent of this inhibition was dependent on the ovary stage of the MO-donor animal, being maximal in MOs from animals in the early stages of ovarian development. Assay of FA O methyltransferase activity, using [3H]FA in the presence of S-adenosyl-l methionine, demonstrated that the enzyme was located in the cytosolic fraction of MOs and was inhibited by incubation of MOs with MO-IH prior to preparation of subcellular fractions. For cytosolic preparations taken from vitellogenic animals, both Vmax and Km were appreciably lower than for those taken from non vitellogenic animals. Conversely, eyestalk ablation of early-vitellogenic animals, which removes the source of MO-IH in vivo, resulted in enhancement of the cytosolic FA O-methyltransferase activity. Although both Vmax and Km show an appreciable increase upon eyestalk ablation, the increased enzyme activity is probably reflected by the fact that Vmax/Km (an approximate indication of kcat) has increased 5-fold. The combined evidence demonstrates that MO-IH inhibits FA O methyltransferase, the enzyme which catalyses the final step of MF biosynthesis in MOs. PMID- 9729473 TI - Intracellular calcium mobilization and phospholipid degradation in sphingosylphosphorylcholine-stimulated human airway epithelial cells. AB - Extracellular sphingosylphosphorylcholine (SPC) caused a remarkable elevation in the intracellular Ca2+ concentration ([Ca2+]i) in immortalized human airway epithelial cells (CFNP9o-). An increase in total inositol phosphates formation was determined; however, the dose responses for [Ca2+]i elevation and inositol phosphates production were slightly different and, furthermore, PMA and pertussis toxin almost completely inhibited [Ca2+]i mobilization by SPC, whereas inositol phosphates production was only partially reduced. The possible direct interaction of SPC with Ca2+ channels of intracellular stores was determined by experiments with permeabilized cells, where SPC failed to evoke Ca2+ release, whereas lysophosphatidic acid was shown to be effective. The level of phosphatidic acid was increased by SPC only in the presence of AACOCF3, a specific inhibitor of phospholipase A2 (PLA2) and blocked by both pertussis toxin and R59022, an inhibitor of diacylglycerol kinase. R59022 enhanced diacylglycerol production by SPC and also significantly reduced [Ca2+]i mobilization. Only polyunsaturated diacylglycerol and phosphatidic acid were generated by SPC. Lastly, SPC caused stimulation of arachidonic acid release, indicating the involvement of PLA2. Taken together, these data suggest that, after SPC stimulation, phospholipase C derived diacylglycerol is phosphorylated by a diacylglycerol kinase to phosphatidic acid, which is further hydrolysed by PLA2 activity to arachidonic and lysophosphatidic acids. We propose that lysophosphatidic acid might be the intracellular messenger able to release Ca2+ from internal stores. PMID- 9729475 TI - Leishmania major parasites express cyclophilin isoforms with an unusual interaction with calcineurin. AB - The immunosuppressive effects of the fungal metabolite cyclosporin A (CsA) are mediated primarily by binding to cyclophilins (Cyps). The resulting CsA-Cyp complex inhibits the Ca2+-regulated protein phosphatase calcineurin and down regulates signal transduction events. Previously we reported that CsA is a potent inhibitor of infections transmitted by the human pathogenic protozoan parasite Leishmania major in vitro and in vivo, but does not effect the extracellular growth of L. major itself. It is unknown how L. major exerts this resistance to CsA. Here we report that a major Cyp, besides additional isoforms with the same N terminal amino acid sequence, was expressed in L. major. The cloned and sequenced gene encodes a putative 174-residue protein called L. major Cyp 19 (LmCyp19). The recombinant LmCyp19 exhibits peptidyl-prolyl cis/trans isomerase activity with a substrate specificity and an inhibition by CsA that are characteristic of other eukaryotic Cyps. To determine whether calcineurin is involved in the discrimination of the effects of CsA we also examined the presence of a parasitic calcineurin and tested the interaction with Cyps. Despite the expression of functionally active calcineurin by L. major, neither LmCyp19 nor other L. major Cyps bound to its own or mammalian calcineurin. The amino acid sequence of most Cyps includes an essential arginine residue around the calcineurin-docking side. In LmCyp19 this is replaced by an asparagine residue. This exchange and additional charged residues are apparently responsible for the lack of LmCyp19 interaction with calcineurin. These observations indicate that resistance of L. major to CsA in vitro is mediated by the lack of complex formation with calcineurin despite CsA binding by parasitic Cyp. PMID- 9729476 TI - Effects of ethanol on mitogen-activated protein kinase and stress-activated protein kinase cascades in normal and regenerating liver. AB - To understand the mechanisms by which ethanol inhibits hepatocyte proliferation, we studied the effects of ethanol on p42/44 mitogen-activated protein kinase (MAPK), p38 mitogen-activated protein kinase (p38 MAPK) and c-Jun N-terminal kinase (JNK) in normal and regenerating rat liver. Treatment of rat hepatocytes with 100 mM ethanol in vitro for 16 h prolonged the activation of p42/44 MAPK and p38 MAPK induced by various agonists. Such treatment also increased basal JNK activity, but did not potentiate or prolong agonist-induced JNK activation. Ethanol potentiation of the activation of p42/44 MAPK was abolished by pertussis toxin. In contrast, chronic ethanol consumption in vivo inhibited the activation of p42/44 MAPK, p38 MAPK and JNK induced either by partial hepatectomy or by various agonists. However, both acute and chronic ethanol inhibited hepatocyte proliferation induced by insulin and epidermal growth factor. A selective inhibitor of p42/44 MAPK partially prevented the inhibition of hepatocyte proliferation caused by acute, but not by chronic, ethanol exposure, whereas a selective inhibitor of p38 MAPK further inhibited hepatocyte proliferation under both conditions. These data suggest that acute and chronic ethanol inhibit hepatocyte proliferation by different mechanisms. The effect of acute ethanol may be related to the prolongation of p42/44 MAPK activation, whereas inhibition of hepatocyte proliferation by chronic ethanol may be due to inhibition of p38 MAPK activation. PMID- 9729477 TI - A novel family of ubiquitin-specific proteases in chick skeletal muscle with distinct N- and C-terminal extensions. AB - We have recently identified a cDNA for a ubiquitin-specific protease (UBP), UBP41, that encodes the smallest functional UBP identified to date, using an Escherichia coli-based in vivo screening method. In the present study we isolated highly related cDNAs encoding a new family of UBP enzymes, named UBP46, UBP52 and UBP66. These UBPs have virtually identical catalytic domains spanning the sequence of UBP41 between the active-site Cys and the His box (95% identity). However, they possess distinct N- and/or C-terminal extensions. Moreover, they are more closely related to each other than to any other members of the UBP family. Thus these chick UBPs must define a novel family of de-ubiquitinating enzymes and should represent the first example among the UBP family enzymes, whose multiplicity is achieved by variation in their N- and C-terminal extensions. The chick UBPs were expressed in E. coli, and purified from the cells to apparent homogeneity using 125I-labelled ubiquitin-alphaNH-MHISPPEPESEEEEEHYC as a substrate. Each of the purified UBP46, UBP52 and UBP66 enzymes behaved as proteins of similar sizes under both denaturing and non-denaturing conditions, suggesting that all of them consist of a single polypeptide chain. The UBP enzymes cleaved the C-terminus of the ubiquitin moiety in natural and engineered fusions irrespective of their sizes and thus are active against ubiquitin-beta galactosidase as well as a ubiquitin C-terminal extension protein of 80 amino acids. All UBPs except UBP66 released free ubiquitin from poly-His-tagged di ubiquitin. However, the isopeptidase activity for hydrolysing polyubiquitinated lysozyme conjugates was not detected from these UBPs, which makes these UBPs distinct from UBP41. These results suggest that the chick UBPs may play an important role in production of free ubiquitin from linear polyubiquitin chains and of certain ribosomal proteins from ubiquitin fusion proteins. PMID- 9729478 TI - MUC5B is a major gel-forming, oligomeric mucin from human salivary gland, respiratory tract and endocervix: identification of glycoforms and C-terminal cleavage. AB - Mucins from human whole saliva, as well as from respiratory- and cervical-tract secretions, were subjected to density-gradient centrifugation in CsCl/0.5 M guanidinium chloride. A polydisperse population of MUC5B mucins was demonstrated in all samples using anti-peptide antisera (LUM5B-2, LUM5B-3 and LUM5B-4) raised against sequences within the MUC5B mucin. The sequences recognized by the LUM5B-2 and LUM5B-3 antisera are located within the domains flanking the highly glycosylated regions of MUC5B, and reduction increased the reactivity with these antibodies, suggesting that the epitopes are partially shielded and that these regions are folded and stabilized by disulphide bonds. Rate-zonal centrifugation before and after reduction showed MUC5B to be a large oligomeric mucin composed of disulphide-linked subunits. In saliva and respiratory-tract secretions, populations of MUC5B mucins with different charge densities were identified by ion-exchange HPLC, suggesting the presence of MUC5B 'glycoforms'. In trachea, the F2 monoclonal antibody against the sulpho-Lewis C structure reacted preferentially with the later-to-be-eluted populations. An antibody (LUM5B-4) recognizing a sequence in the C-terminal domain of MUC5B identified, after reduction, the mucin subunits as well as smaller fragments, suggesting that some of the MUC5B mucins are cleaved within the C-terminal domain. Immunohistochemistry revealed that MUC5B is produced by cells dispersed throughout the human submandibular and sublingual glands, in the airway submucosal glands as well as the goblet cells, and in the epithelium and glands of the endocervix. The F2 antibody stained a subpopulation of the MUC5B-producing cells in the airway submucosal glands, suggesting that different cells may produce different glycoforms of MUC5B in this tissue. PMID- 9729479 TI - Isolation and identification of metallothionein isoforms (MT-1 and MT-2) in the rat testis. AB - It has been a long-lasting controversial issue as to whether or not the male genital organs, such as the testis and prostate, contain metallothioneins (MTs), a group of cysteine-rich heavy-metal-binding proteins that play a role in detoxifying heavy metals such as cadmium (Cd). Earlier studies reported that the rodent testis lacks MTs and concluded that this is why the testis is very susceptible to Cd, although other indirect experimental evidence suggests that MTs are present in this organ. A deficiency of MTs in the testis was originally suspected on the basis of amino acid composition analysis, since MT-like proteins isolated as Cd-binding proteins did not have a characteristic MT structure. In the present study, we demonstrate that the rat testis indeed expresses Cd-binding proteins with sequences identical to those previously described for MT-1 and MT 2, the major isoforms. To confirm that MT-1 and MT-2 are present in the rat testis, we purified and isolated Cd-binding proteins by homogenization using Cd containing buffer, followed by sequential purification using Sephadex G-75 gel filtration chromatography and anion HPLC column chromatography, which yielded Cd binding protein-1 (Cd-BP-1) and -2 (Cd-BP-2). After pyridylethylation, Cd-BP-1 and Cd-BP-2 were subjected to specific protein fragmentation by acids and endopeptidases, which revealed that these Cd-binding proteins have the same primary structures as MT-1 and MT-2 respectively. Thus we believe that the present results clearly resolve the long-standing debate about the presence of MTs in the testis, at least in the rodent. PMID- 9729480 TI - Heterologously expressed inner lipoyl domain of dihydrolipoyl acetyltransferase inhibits ATP-dependent inactivation of pyruvate dehydrogenase complex. Identification of important amino acid residues. AB - The activity of the pyruvate dehydrogenase multienzyme complex (PDC), which catalyses the oxidation of pyruvate to acetyl-CoA within the mitochondrion, is diminished in animal models of diabetes. Studies with purified PDC components have suggested that the kinases responsible for inactivating the decarboxylase catalytic subunits of the complex are most efficient in their regulatory role when they are bound to dihydrolipoyl acetyltransferase (E2) subunits, which form the structural core of the complex. We report that the addition of an exogenous E2 subdomain (inner lipoyl domain) to an intact PDC inhibits ATP-dependent inactivation of the complex. By combining molecular modelling, site-directed mutagenesis and biophysical characterizations, we have also identified two amino acid residues in this subdomain (Ile229 and Phe231) that largely determine the magnitude of this effect. PMID- 9729482 TI - ATP-dependent transport of reduced glutathione in yeast secretory vesicles. AB - Turnover of cellular reduced glutathione (GSH) is accomplished predominantly by export into the extracellular space; however, the plasma membrane transport mechanisms that mediate GSH efflux are not well characterized. The present study examined GSH transport using secretory vesicles isolated from the sec6-4 mutant strain of Saccharomyces cerevisiae. In contrast with studies in mammalian membrane vesicles, GSH transport in yeast secretory vesicles was mediated largely by an ATP-dependent, low-affinity pathway (Km 19+/-5 mM). ATP-dependent [3H]GSH transport was cis-inhibited by substrates of the yeast YCF1 transporter, including sulphobromophthalein, glutathione S-conjugates and the alkaloid verapamil, and was competitively inhibited by S-(2, 4-dinitrophenyl)glutathione (DNP-SG). Similarly, GSH competitively inhibited ATP-dependent [3H]DNP-SG transport, with a Ki of 18+/-2 mM, but had no effect on ATP-dependent [3H]taurocholate transport. ATP-dependent GSH transport was not affected by either membrane potential or pH-gradient uncouplers, but was inhibited by 4, 4' di-isothiocyanatostilbene-2,2'-disulphonate, probenecid and sulphinpyrazone, which are inhibitors of mrp1 and mrp2, mammalian homologues of the yeast YCF1 transporter. Western blot analysis of the secretory vesicle membrane fraction confirmed the presence of Ycf1p. These results provide the first direct evidence for low-affinity, ATP-dependent transport of GSH, and demonstrate that this ATP dependent pathway displays kinetic characteristics similar to those of the yeast YCF1 transporter. PMID- 9729481 TI - Mechanism for basal expression of rat mitochondrial branched-chain-2-oxo-acid dehydrogenase kinase [corrected]. AB - The rat branched-chain-2-oxo-acid dehydrogenase (BCOD) kinase mRNA is transcribed from a TATA-less promoter that has GC-rich sequences and two putative Sp1 binding sites near the transcription start site. We demonstrated previously that the 5' region of the kinase gene, base pairs -128 to +264, contained promoter activity when assayed using luciferase as a reporter (Huang and Chuang (1996) Biochem. J. 313, 603-609). To define DNA elements required for efficient expression of the kinase gene, nested deletion constructs of the above promoter region fused with a luciferase reporter gene were transfected into cultured H4IIE (hepatoma) and NRK 52E (kidney) cells. The results showed that the region between nucleotides -58 and +21 was indispensable for the kinase basal promoter activity. Methylation interference and mutagenesis-promoter assays identified nucleotides -50 to -40 (ACAACTCCCA) as cis-acting DNA sequences that are required for nuclear protein binding and efficient promoter activity. Gel-supershift analysis with anti-Sp1 antibody suggested that the nuclear protein capable of binding to the -58 oligonucleotide (bp -58 to -34) was immunologically related to the Sp1 protein. The -58 oligonucleotide formed a DNA-protein complex with recombinant Sp1 protein with an affinity approximately ten-fold lower than that of the consensus Sp1 oligonucleotide. Co-transfection of the Sp1 expression plasmid and the -58 promoter construct into Drosophila Schneider cells revealed that Sp1 contributed to the kinase basal promoter activity by binding to the non-consensus site in the -58 region. Deletion of two consensus Sp1 binding sites (bases -150 to -140 and bases +29 to +38) in the kinase gene did not affect the basal promoter activity. Therefore binding of Sp1 or Sp1-like proteins to the above single non-consensus Sp1 sequence in the -58 region plays a major role of transactivating basal expression of the BCOD kinase. PMID- 9729484 TI - Signals transmitted along retinal axons in Drosophila: Hedgehog signal reception and the cell circuitry of lamina cartridge assembly. AB - The arrival of retinal axons in the brain of Drosophila triggers the assembly of glial and neuronal precursors into a 'neurocrystalline' array of lamina synaptic 'cartridges'. Hedgehog, a secreted protein, is an inductive signal delivered by retinal axons for the initial steps of lamina differentiation. In the development of many tissues, Hedgehog acts in a signal relay cascade via the induction of secondary secreted factors. Here we show that lamina neuronal precursors respond directly to Hedgehog signal reception by entering S-phase, a step that is controlled by the Hedgehog-dependent transcriptional regulator Cubitus interruptus. The terminal differentiation of neuronal precursors and the migration and differentiation of glia appear to be controlled by other retinal axon-mediated signals. Thus retinal axons impose a program of developmental events on their postsynaptic field utilizing distinct signals for different precursor populations. PMID- 9729485 TI - Two distinct mechanisms for differential positioning of gene expression borders involving the Drosophila gap protein giant. AB - We have combined genetic experiments and a targeted misexpression approach to examine the role of the gap gene giant (gt) in patterning anterior regions of the Drosophila embryo. Our results suggest that gt functions in the repression of three target genes, the gap genes Kruppel (Kr) and hunchback (hb), and the pair rule gene even-skipped (eve). The anterior border of Kr, which lies 4-5 nucleus diameters posterior to nuclei that express gt mRNA, is set by a threshold repression mechanism involving very low levels of gt protein. In contrast, gt activity is required, but not sufficient for formation of the anterior border of eve stripe 2, which lies adjacent to nuclei that express gt mRNA. We propose that gt's role in forming this border is to potentiate repressive interaction(s) mediated by other factor(s) that are also localized to anterior regions of the early embryo. Finally, gt is required for repression of zygotic hb expression in more anterior regions of the embryo. The differential responses of these target genes to gt repression are critical for the correct positioning and maintenance of segmentation stripes, and normal anterior development. PMID- 9729483 TI - Prodigiosins uncouple lysosomal vacuolar-type ATPase through promotion of H+/Cl- symport. AB - We reported previously [Kataoka, Muroi, Ohkuma, Waritani, Magae, Takatsuki, Kondo, Yamasaki and Nagai (1995) FEBS Lett. 359, 53-59] that prodigiosin 25-C (one of the red pigments of the prodigiosin group produced by micro-organisms like Streptomyces and Serratia) uncoupled vacuolar H+-ATPase, inhibited vacuolar acidification and affected glycoprotein processing. In the present study we show that prodigiosin, metacycloprodigiosin and prodigiosin 25-C, all raise intralysosomal pH through inhibition of lysosomal acidification driven by vacuolar-type (V-)ATPase without inhibiting ATP hydrolysis in a dose-dependent manner with IC50 values of 30-120 pmol/mg of protein. The inhibition against lysosomal acidification was quick and reversible, showing kinetics of simple non competitive (for ATP) inhibition. However, the prodigiosins neither raised the internal pH of isolated lysosomes nor showed ionophoric activity against H+ or K+ at concentrations where they strongly inhibited lysosomal acidification. They required Cl- for their acidification inhibitory activity even when driven in the presence of K+ and valinomycin, suggesting that their target is not anion (chloride) channel(s). In fact, the prodigiosins inhibited acidification of proteoliposomes devoid of anion channels that were reconstituted from lysosomal vacuolar-type (V-)ATPase and Escherichia coli phospholipids. However, they did not inhibit the formation of an inside-positive membrane potential driven by lysosomal V-ATPase. Instead, they caused quick reversal of acidified pH driven by lysosomal V-ATPase and, in acidic buffer, produced quick acidification of lysosomal pH, both only in the presence of Cl-. In addition, they induced swelling of liposomes and erythrocytes in iso-osmotic ammonium salt of chloride but not of gluconate, suggesting the promotion of Cl- entry by prodigiosins. These results suggest that prodigiosins facilitate the symport of H+ with Cl- (or exchange of OH- with Cl-) through lysosomal membranes, resulting in uncoupling of vacuolar H+-ATPase. PMID- 9729487 TI - The Arabidopsis thaliana gametophytic mutation gemini pollen1 disrupts microspore polarity, division asymmetry and pollen cell fate. AB - Pollen development and male gametogenesis are critically dependent upon cell polarization leading to a highly asymmetric cell division termed pollen mitosis I. A mutational approach was adopted in Arabidopsis thaliana to identify genes involved these processes. Four independent gemini pollen mutants were isolated which produce divided or twin-celled pollen. The gemini pollen1 mutant was characterized in detail and shown to act gametophytically resulting in reduced transmission through both sexes. gemini pollen1 showed an incompletely penetrant phenotype resulting in equal, unequal and partial divisions at pollen mitosis I. The division planes in gemini pollen1 were shown to be aligned with the polar axis (as in wild type) and evidence was obtained for incomplete nuclear migration, which could account for altered division symmetry. gemini pollen1 also showed division phenotypes consistent with spatial uncoupling of karyokinesis and cytokinesis suggesting that GEMINI POLLEN1 may be required for the localization of phragmoplast activity. Cell fate studies showed that in both equal and unequal divisions a vegetative cell marker gene was activated in both daughter cells. Daughter cells with a range of intermediate or hybrid vegetative/generative cell fates suggests that cell fate is quantitatively related to cell size. The potential mode of action of GEMINI POLLEN1 and its effects on cell fate are discussed in relation to proposed models of microspore polarity and cell fate determination. PMID- 9729486 TI - BMPs mediate lateral inhibition at successive stages in feather tract development. AB - The spacing of feather buds in a tract is thought to arise from the interaction between an inducing signal from the dermis and an inhibitory signal generated in the nascent buds. Local BMP-2 expression in the ectoderm precedes the formation of the ectodermal placodes, which are the first morphological sign of bud differentiation. We have altered the activity of BMP-2 or BMP-4 in the ectoderm of the feather field by expressing them or their inhibitor noggin using retroviral vectors. These experiments demonstrate that BMP-2 is necessary and sufficient to mediate the lateral inhibition that positions buds in a tract. After buds are initiated, BMP-2 and BMP-4 continue to inhibit the adoption of bud fates and help to specify the size and shape of the bud. They may do so in part by their regulation of Fgf receptor expression in both the ectoderm and dermis. Additional insights into pattern formation in the feather bud can be inferred from the effects of altered BMP activity on bud morphogenesis. PMID- 9729488 TI - A Serum Response Factor homolog is required for spore differentiation in Dictyostelium. AB - A homolog of the Serum Response Factor (SRF) has been isolated from Dictyostelium discoideum and its function studied by analyzing the consequences of its gene disruption. The MADS-box region of Dictyostelium SRF (DdSRF) is highly conserved with those of the human, Drosophila and yeast homologs. srfA is a developmentally regulated gene expressed in prespore and spore cells. This gene plays an essential role in sporulation as its disruption leads to abnormal spore morphology and loss of viability. The mutant spores were round and cellulose deposition seemed to be partially affected. Initial prestalk and prespore cell differentiation did not seem to be compromised in the mutant since the expression of several cell-type-specific markers were found to be unaffected. However, the mRNA level of the spore marker spiA was greatly reduced. Activation of the cAMP dependent protein kinase (PKA) by 8-Br-cAMP was not able to fully bypass the morphological defects of srfA- mutant spores, although this treatment induced spiA mRNA expression. Our results suggest that DdSRF is required for full maturation of spores and participates in the regulation of the expression of the spore-coat marker spiA and probably other maturation genes necessary for proper spore cell differentiation. PMID- 9729489 TI - Developmental remodeling and shortening of the cardiac outflow tract involves myocyte programmed cell death. AB - The embryonic outflow tract is a simple tubular structure that connects the single primitive ventricle with the aortic sac and aortic arch arteries. This structure undergoes a complex sequence of morphogenetic processes to become the portion of the heart that aligns the right and left ventricles with the pulmonary artery and aorta. Abnormalities of the outflow tract are involved in many clinically significant congenital cardiac defects; however, the cellular and molecular processes governing the development of this important structure are incompletely understood. Histologic and tissue-tagging studies indicate that the outflow tract tissues compact and are incorporated predominantly into a region of the right ventricle. The hypothesis tested in the current study was that cell death or apoptosis in the muscular portion of the outflow tract is an important cellular mechanism for outflow tract shortening. The tubular outflow tract myocardium was specifically marked by infecting myocytes of the chicken embryo heart with a recombinant replication-defective adenovirus expressing beta galactosidase (beta-gal) under the control of the cytomegalovirus promoter. Histochemical detection of the beta -gal-labeled outflow tract myocytes revealed that the tubular structure shortened to become a compact ring at the level of the pulmonic infundibulum over several days of development (stages 25-32, embryonic days 4-8). The appearance of apoptotic cardiomyocytes was correlated with OFT shortening by two histologic assays, TUNEL labeling of DNA fragments and AnnexinV binding. The rise and fall in the number of apoptotic myocytes detected by histologic analyses paralleled the change in activity levels of Caspase-3, a protease in the apoptotic cascade, measured in outflow tract homogenates. These results suggest that the elimination of myocytes by programmed cell death is one mechanism by which the outflow tract myocardium remodels to form the proper connection between the ventricular chambers and the appropriate arterial trunks. PMID- 9729490 TI - Generation of multiple antagonistic domains along the proximodistal axis during Drosophila leg development. AB - homothorax (hth) is a Drosophila member of the Meis family of homeobox genes. hth function is required for the nuclear localization of the Hox cofactor Extradenticle (EXD). We show here that there is also a post-transcriptional control of HTH by exd: exd activity is required for the apparent stability of the HTH protein. In leg imaginal discs, hth expression is limited to the domain of exd function and this domain is complementary to the domain in which the Wingless (WG) and Decapentaplegic (DPP) signals are active. We demonstrate that WG and DPP act together through their targets Distal-less (Dll) and dachshund (dac) to restrict hth expression, and therefore EXD's nuclear localization, to the most proximal regions of the leg disc. Furthermore, there is a reciprocal repression exerted by HTH on these and other DPP and WG downstream targets that restricts their expression to non-hth-expressing cells. Thus, there exists in the leg disc a set of mutually antagonistic interactions between proximal cells, which we define as those that express hth, and distal cells, or those that do not express hth. In addition, we show that dac negatively regulates Dll. We suggest that these antagonistic relationships help to convert the WG and DPP activity gradients into discreet domains of gene expression along the proximodistal axis. PMID- 9729491 TI - Prx1 and Prx2 in skeletogenesis: roles in the craniofacial region, inner ear and limbs. AB - Prx1 and Prx2 are closely related paired-class homeobox genes that are expressed in very similar patterns predominantly in mesenchyme. Prx1 loss-of-function mutants show skeletal defects in skull, limbs and vertebral column (Martin, J. F., Bradley, A. and Olson, E. N. (1995) Genes Dev. 9, 1237-1249). We report here that mice in which Prx2 is inactivated by a lacZ insertion had no skeletal defects, whereas Prx1/Prx2 double mutants showed many novel abnormalities in addition to an aggravation of the Prx1 single mutant phenotype. We found defects in external, middle and inner ear, reduction or loss of skull bones, a reduced and sometimes cleft mandible, and limb abnormalities including postaxial polydactyly and bent zeugopods. A single, or no incisor was present in the lower jaw, and ectopic expression of Fgf8 and Pax9 was found medially in the mandibular arch. A novel method to detect &bgr ;-galactosidase activity in hydroxyethylmethacrylate sections allowed detailed analysis of Prx2 expression in affected structures. Our results suggest a role for Prx genes in mediating epitheliomesenchymal interactions in inner ear and lower jaw. In addition, Prx1 and Prx2 are involved in interactions between perichondrium and chondrocytes that regulate their proliferation or differentiation in the bones of the zeugopods. PMID- 9729492 TI - CLAVATA2, a regulator of meristem and organ development in Arabidopsis. AB - Mutations at the CLAVATA2 (CLV2) locus of Arabidopsis result in enlarged shoot and flower meristems, as well as alterations in the development of the gynoecia, flower pedicels, and stamens. The shoot and flower meristem phenotypes of clv2 mutants are similar to weak clv1 and clv3 mutants. We present genetic analysis that CLV2 may function in the same pathway as CLV1 and CLV3 in the regulation of meristem development, but function separately in the regulation of organ development. We also present evidence that clv2 phenotypes are altered when the mutants are grown under short-day light conditions. These alterations include flower-to-shoot transformations, as well as a nearly complete suppression of the flower phenotypes, indicating that the requirement for CLV2 changes in response to different physiological conditions. The stm-1 mutation dominantly suppresses clv2, and clv2 mutations suppress the strong stm-1 allele, but not the weak stm-2 allele. PMID- 9729493 TI - Commissureless endocytosis is correlated with initiation of neuromuscular synaptogenesis. AB - We show that the Commissureless (COMM) transmembrane protein is required at neuromuscular synaptogenesis. All muscles in the Drosophila embryo express COMM during the period of motoneuron-muscle interaction. It is endocytosed into muscles before synaptogenesis. In comm loss-of-function mutants, motoneuron growth cones fail to initiate synaptogenesis at target muscles. This stall phenotype is rescued by supplying wild-type COMM to the muscles. Cytoplasmically truncated COMM protein fails to internalize. Expressing this mutant protein in muscles phenocopies the synaptogenesis defects of comm mutants. Thus, synaptogenesis initiation is positively correlated with endocytosis of COMM in postsynaptic muscle cells. We propose that COMM is an essential part of the dynamic cell surface remodeling needed by postsynaptic cells in coordinating synaptogenesis initiation. PMID- 9729494 TI - The Pristionchus HOX gene Ppa-lin-39 inhibits programmed cell death to specify the vulva equivalence group and is not required during vulval induction. AB - In the two nematode species Caenorhabditis elegans and Pristionchus pacificus the vulva equivalence group in the central body region is specified by the Hox gene lin-39. C. elegans lin-39 mutants are vulvaless and the vulval precursor cells fuse with the surrounding hypodermis, whereas in P. pacificus lin-39 mutants the vulval precursor cells die by apoptosis. Mechanistically, LIN-39 might inhibit non-vulval fate (cell fusion in C. elegans, apoptosis in P. pacificus), promote vulval fate or do both. To study the mechanism of lin-39 function, we isolated P. pacificus cell death mutants and identified mutations in ced-3. Surprisingly, P. pacificus ced-3; lin-39 double mutants form a functional vulva in the absence of LIN-39 activity. Thus, in P. pacificus lin-39 specifies the vulva equivalence group by inhibiting programmed cell death. Furthermore, these data reveal an important difference in a later function of lin-39 between the two species. In C. elegans, LIN-39 specifies vulval cell fates in response to inductive RAS signaling, and in P. pacificus LIN-39 is not required for vulval induction. Thus, the comparative analysis indicates that lin-39 has distinct functions in both species although the gene is acting in a homologous developmental system. PMID- 9729495 TI - Dissecting the roles of the Drosophila EGF receptor in eye development and MAP kinase activation. AB - A new conditional Egfr allele was used to dissect the roles of the receptor in eye development and to test two published models. EGFR function is necessary for morphogenetic furrow initiation, is not required for establishment of the founder R8 cell in each ommatidium, but is necessary to maintain its differentiated state. EGFR is required subsequently for recruitment of all other neuronal cells. The initial EGFR-dependent MAP kinase activation occurs in the furrow, but the active kinase (dp-ERK) is observed only in the cytoplasm for over 2 hours. Similarly, SEVENLESS-dependent activation results in cytoplasmic appearance of dp ERK for 6 hours. These results suggest an additional regulated step in this pathway and we discuss models for this. PMID- 9729496 TI - The T-box transcription factor Brachyury regulates expression of eFGF through binding to a non-palindromic response element. AB - Brachyury is a member of the T-box gene family and is required for formation of posterior mesoderm and notochord during vertebrate development. The ability of Brachyury to activate transcription is essential for its biological function, but nothing is known about its target genes. Here we demonstrate that Xenopus Brachyury directly regulates expression of eFGF by binding to an element positioned approximately 1 kb upstream of the eFGF transcription start site. This site comprises half of the palindromic sequence previously identified by binding site selection and is also present in the promoters of the human and mouse homologues of eFGF. PMID- 9729497 TI - Cell-type-specific rescue of myosin function during Dictyostelium development defines two distinct cell movements required for culmination. AB - Mutant Dictyostelium cells lacking any of the component polypeptides of myosin II exhibit developmental defects. To define myosin's role in establishing Dictyostelium's developmental pattern, we have rescued myosin function in a myosin regulatory light chain null mutant (mlcR-) using cell-type-specific promoters. While mlcR- cells fail to progress beyond the mound stage, expression of RLC from the prestalk promoter, ecmA, produces culminants with normal stalks but with defects in spore cell localization. When GFP-marked prestalk and prespore cells expressing ecmA-RLC are mixed with wild-type cells, the mislocalization of prestalk cells, but not prespore cells, is rescued. Time-lapse video recording of ecmA-RLC cells showed that the posterior prespore zone failed to undergo a contraction important for the upward movement of prespore cells. Prespore cells marked with green fluorescent protein (GFP) failed to move toward the tip with the spiral motion typical of wild type. In contrast, expression of RLC in prespore cells using the psA promoter produced balloon-like structures reminiscent of sorocarps but lacking stalks. GFP-labeled prespore cells showed a spiral movement toward the top of the structures. Expression of RLC from the psA promoter restores the normal localization of psA-GFP cells, but not ecmA-GFP cells. These results define two distinct, myosin-dependent movements that are required for establishing a Dictyostelium fruiting body: stalk extension and active movement of the prespore zone that ensures proper placement of the spores atop the stalk. The approach used in these studies provides a direct means of testing the role of cell motility in distinct cell types during a morphogenetic program. PMID- 9729498 TI - VEGF mediates angioblast migration during development of the dorsal aorta in Xenopus. AB - Angioblasts are precursor cells of the vascular endothelium which organize into the primitive blood vessels during embryogenesis. The molecular mechanisms underlying patterning of the embryonic vasculature remain unclear. Mutational analyses of the receptor tyrosine kinase flk-1 and its ligand vascular endothelial growth factor, VEGF, indicate that these molecules are critical for vascular development. Targeted ablation of the flk-1 gene results in complete failure of blood and vascular development (F. Shalaby et al. (1995) Nature 376, 62-66), while targeted ablation of the VEGF gene results in gross abnormalities in vascular patterning (P. Carmeliet et al. (1996) Nature 380, 435-439; N. Ferrara et al. (1996) Nature 380, 439-442). Here we report a role for VEGF in patterning the dorsal aorta of the Xenopus embryo. We show that the diffusible form of VEGF is expressed by the hypochord, which lies at the embryonic midline immediately dorsal to the location of the future dorsal aorta. We find that, initially, no flk-1-expressing angioblasts are present at this location, but that during subsequent development, angioblasts migrate from the lateral plate mesoderm to the midline where they form a single dorsal aorta. We have demonstrated that VEGF can act as a chemoattractant for angioblasts by ectopic expression of VEGF in the embryo. These results strongly suggest that localized sources of VEGF play a role in patterning the embryonic vasculature. PMID- 9729499 TI - Function and specificity of LIM domains in Drosophila nervous system and wing development. AB - LIM domains are found in a variety of proteins, including cytoplasmic and nuclear LIM-only proteins, LIM-homeodomain (LIM-HD) transcription factors and LIM kinases. Although the ability of LIM domains to interact with other proteins has been clearly established in vitro and in cultured cells, their in vivo function is unknown. Here we use Drosophila to test the roles of the LIM domains of the LIM-HD family member Apterous (Ap) in wing and nervous system development. Using a rescuing assay of the ap mutant phenotype, we have found that the LIM domains are essential for Ap function. Furthermore, expression of LIM domains alone can act in a dominant-negative fashion to disrupt Ap function. The Ap LIM domains can be replaced by those of another family member to generate normal wing structure, but LIM domains are not interchangeable during axon pathfinding of the Ap neurons. This suggests that the Ap LIM domains mediate different protein interactions in different developmental processes, and that LIM domains can participate in conferring specificity of target gene selection. PMID- 9729501 TI - Urea transporters in kidney: molecular analysis and contribution to the urinary concentrating process1. AB - Facilitated urea transporters (UTs) are responsible for urea accumulation in the renal inner medulla of the mammalian kidney and therefore play a central role in the urinary concentrating process. Recently, the cDNAs encoding three members of the UT family, UT1, UT2, and UT3 have been cloned. These transporters are expressed in different structures of the mammalian kidney. In rat, UT1 resides in the apical membrane of terminal inner medullary collecting ducts, where it mediates vasopressin-regulated urea reabsorption. UT2 and UT3 are located in descending thin limbs of Henle's loop and descending vasa recta, respectively, and participate in urinary recycling processes, which minimize urea escape from the inner medulla. UT1 and UT2 are regulated independently and respond differently to changes in dietary protein content and hydration state. Identification and characterization of these urea transporters advances our understanding of the molecular basis and regulation of the urinary concentrating mechanism. PMID- 9729502 TI - Roles of Na-K-2Cl and Na-Cl cotransporters and ROMK potassium channels in urinary concentrating mechanism. AB - The cloning of the apical transport proteins involved in NaCl absorption in the thick ascending limb of Henle and the distal convoluted tubule has ushered in a new era that promises to provide molecular details of the urinary concentrating and diluting processes. We now have at hand tools to examine the regulation of these proteins that can affect the concentrating and diluting capacity. PMID- 9729503 TI - Cellular mechanisms of aquaporin trafficking. AB - Aquaporins (AQPs) are a family of functionally important water channel proteins that are of special cell biological interest because of their diverse intracellular targeting and trafficking properties. AQPs have been found in many different cells and tissues. This short review summarizes recent work that addresses the regulation of AQP2 trafficking in response to vasopressin. PMID- 9729504 TI - Long-term regulation of urinary concentrating capacity. AB - Urinary concentrating capacity is regulated in part by a long-term adaptational process involving changes in the absolute abundance of the aquaporin-2 water channel in collecting duct cells. Alterations in aquaporin-2 abundance play key roles in the pathophysiology of several water balance disorders. Escape from the antidiuretic action of vasopressin, e.g. in the syndrome of inappropriate antidiuretic hormone secretion, involves a selective downregulation of aquaporin 2 expression. Excessive water retention causing hyponatremia in volume-expanded states such as congestive heart failure appears to be due in part to a failure of this escape mechanism. PMID- 9729505 TI - Functional and molecular characterization of luminal and basolateral Cl-/HCO-3 exchangers of rat thick limbs. AB - Cl-/HCO-3 exchange was measured in luminal (LMV) and basolateral (BLMV) membrane vesicles purified from rat medullary thick ascending limb (MTAL). Cl-/HCO-3 exchange in BLMV and LMV was inhibited by DIDS, with respective IC50 values of 3.2 +/- 0.9 and 15.2 +/- 5.2 microM, whereas Cl- conductances were DIDS insensitive. At constant external pH, BLMV 36Cl-/HCO-3 and 36Cl-/Cl- exchanges exhibited a sigmoidal pattern of activation as internal pH (pHi) increased from 6.1 to 8.0, whereas LMV 36Cl-/Cl- exchange was unchanged between pHi 6.7 and 7.8. The 165-kDa AE2 polypeptide and approximately 115-kDa AE1-related polypeptide were present only in BLMV. In contrast, AE1-related polypeptides of approximately 90 and 95 kDa were present not only in BLMV but also (in variable abundance) in LMV. We conclude that rat MTAL BLMV and LMV express distinct anion exchange activities and distinct sets of AE polypeptides. AE2 (and perhaps AE1) in BLMV likely contribute to HCO-3 absorption. In contrast, LMV exchangers may contribute to NaCl absorption via parallel coupling with the luminal Na+/H+ antiporters and/or may provide negative feedback regulation of HCO-3 absorption. PMID- 9729506 TI - Bradykinin stimulates the ERK-->Elk-1-->Fos/AP-1 pathway in mesangial cells. AB - Among its diverse biological actions, the vasoactive peptide bradykinin (BK) induces the transcription factor AP-1 and proliferation of mesangial cells (S. S. El-Dahr, S. Dipp, I. V. Yosipiv, and W. H. Baricos. Kidney Int. 50: 1850-1855, 1996). In the present study, we examined the role of protein tyrosine phosphorylation and the mitogen-activated protein kinases, ERK1/2,in mediating BK induced AP-1 and DNA replication in cultured rat mesangial cells. BK (10(-9) to 10(-7) M) stimulated a rapid increase in tyrosine phosphorylation of multiple proteins with an estimated molecular mass of 120-130, 90-95, and 44-42 kDa. Immunoblots using antibodies specific for ERK or tyrosine-phosphorylated ERK revealed a shifting of p42 ERK2 to a higher molecular weight that correlated temporally with an increase in tyrosine-phosphorylated ERK2. Genistein, a specific tyrosine kinase inhibitor, prevented the phosphorylation of ERK2 by BK. In-gel kinase assays indicated that BK-induced tyrosine phosphorylation of ERK2 is accompanied by fourfold activation of its phosphotransferase activity toward the substrate PHAS-I (P < 0.05). Furthermore, BK stimulated a 2.5-fold increase (P < 0.05) in phosphorylation of Elk-1, a transcription factor required for growth factor-induced c-fos transcription. In accord with the stimulation of Elk 1 phosphorylation, BK induced c-fos gene expression and the production of Fos/AP 1 complexes. In addition, thymidine incorporation into DNA increased twofold (P < 0. 05) following BK stimulation. Each of these effects was blocked by tyrosine kinase inhibition with genistein or herbimycin A. Similarly, antisense oligodeoxynucleotide targeting of ERK1/2 mRNA inhibited BK-stimulated DNA synthesis. In contrast, protein kinase C inhibition or depletion had no effect on BK-induced c-fos mRNA, AP-1-DNA binding activity, or DNA synthesis. Collectively, these data demonstrate that BK activates the ERK-->Elk-1-->AP-1 pathway and that BK mitogenic signaling is critically dependent on protein tyrosine phosphorylation. PMID- 9729507 TI - Mg2+/Ca2+ sensing inhibits hormone-stimulated Mg2+ uptake in mouse distal convoluted tubule cells. AB - The distal convoluted tubule plays a significant role in renal magnesium conservation. An immortalized mouse distal convoluted tubule (MDCT) cell line has been extensively used to study the cellular mechanisms of magnesium transport in this nephron segment. MDCT cells possess an extracellular polyvalent cation sensing mechanism responsive to Mg2+, Ca2+, and neomycin. The present studies determined the effect of Mg2+/Ca2+ sensing on hormone-mediated cAMP formation and Mg2+ uptake in MDCT cells. MDCT cells were Mg2+ depleted by culturing in Mg2+ free media for 16 h, and Mg2+ uptake was measured by microfluorescence after placing the depleted cells in 1.5 mM MgCl2. The mean rate of Mg2+ uptake was 164 +/- 5 nM/s in control MDCT cells. Activation of Mg2+/Ca2+ sensing with neomycin did not affect basal Mg2+ uptake (155 +/- 5 nM/s). We have previously reported that treatment of MDCT cells with either glucagon or arginine vasopressin (AVP) stimulated Mg2+ entry. In the present studies, the addition of extracellular Mg2+ or Ca2+ inhibited glucagon- and AVP-stimulated cAMP formation and Mg2+ uptake in concentration-dependent manner with half-maximal concentrations of approximately 1.5 and 3.0 mM, respectively. Exogenous cAMP or forskolin stimulated Mg2+ uptake in the presence of Mg2+/Ca2+ sensing activation. We infer from these studies that Mg2+/Ca2+-sensing mechanisms located in the distal convoluted tubule may play a role in control of distal magnesium absorption. PMID- 9729508 TI - PMA and staurosporine affect expression of the PCK gene in LLC-PK1-F+ cells. AB - The addition of phorbol 12-myristate 13-acetate (PMA) to renal LLC-PK1-F+ cells caused a rapid decrease in the level of phosphoenolpyruvate carboxykinase (PCK) mRNA and reversed the stimulatory effects of exposure to acidic medium (pH 6.9, 10 mM HCO-3) or cAMP. In contrast, prolonged treatment with PMA increased the levels of PCK mRNA. The two effects correlated with the membrane translocation and downregulation of the alpha-isozyme of protein kinase C and were blocked by pretreatment with specific inhibitors of protein kinase C. The rapid decrease in PCK mRNA caused by PMA occurred with a half-life (t1/2 = 1 h) that is significantly faster than that measured during recovery from acid medium or following inhibition of transcription (t1/2 = 4 h). The effect of PMA was reversed by staurosporine, which apparently acts by inhibiting a signaling pathway other than protein kinase C. Staurosporine had no effect on the half-life of the PCK mRNA, but it stimulated the activity of a chloramphenicol acetyltransferase gene that was driven by the initial 490 base pairs of the PCK promoter and transiently transfected into LLC-PK1-F+ cells. This effect was additive to that of cAMP, and neither stimulation was reversed by PMA. The stimulatory effect of staurosporine was mapped to the cAMP response element (CRE 1) and P3(II) element of the PCK promoter. The data indicate that, in LLC-PK1-F+ cells, activation of protein kinase C decreases the stability of the PCK mRNA, whereas transcription of the PCK gene may be suppressed by a kinase that is inhibited by staurosporine. PMID- 9729509 TI - Contribution of 20-HETE to the vasodilator actions of nitric oxide in renal arteries. AB - The present study examined the contribution of elevations in cGMP versus inhibition of cytochrome P-4504A enzymes and the production of the vasoconstrictor 20-hydroxyeicosatetraenoic acid (20-HETE) to the vasodilator actions of NO in renal arterioles. The NO donor sodium nitroprusside (SNP) at 10( 5), 10(-4), and 10(-3) M reduced the production of 20-HETE in microsomes prepared from renal arterioles to 80 +/- 2, 43 +/- 5, and 7 +/- 1% of control, respectively (n = 4). In other experiments, the vasodilator response to SNP (10( 7) to 10(-3) M) was examined in rat renal interlobular arteries (<90 micron ID), preconstricted with phenylephrine (1 microM) under control conditions and after blockade of the cGMP and P-4504A pathways. Inhibition of guanylyl cyclase with 1H [1,2, 4]oxadiazole[4,3-a]quinoxalin-1-one (ODQ) (10 microM, n = 6) or of cGMP dependent protein kinase with 8R,9S, 11S-(-)-9-methoxy-carbamyl-8-methyl-2,3,9,10 tetrahydro-8, 11-epoxy-1H,8H,11H-2,7b,11a-trizadibenzo-(a,g)-cycloocta-(c, d, e) trinden-1-one (KT-5823, 1 microM; n = 5) attenuated the vasodilator response to SNP by 26 and 30%, respectively. In contrast, inhibition of the endogenous production of 20-HETE with a suicide substrate, irreversible inhibitor [17 octadecynoic acid (17-ODYA), 1 microM, n = 5], or a selective, competitive inhibitor of 20-HETE formation (dibromo-dodecenyl-methylsulfimide, 25 microM, n = 5) markedly impaired the vasodilator response to SNP by 76 and 78%, respectively. Similarly, when 20-HETE levels were fixed at 100 nM (n = 6), the response to SNP was attenuated by 73%. Blockade of both pathways with ODQ and 17-ODYA completely abolished the response to SNP (n = 6). These results indicate that the vasodilator response to NO is largely cGMP independent and that inhibition of 20 HETE formation contributes to the cGMP-independent effects of NO in the renal microcirculation. PMID- 9729510 TI - Immunolocalization of the Na+/H+ exchanger isoform NHE2 in rat kidney. AB - Four Na+/H+ exchangers (NHE1 to NHE4) have been detected in the kidney. Renal NHE2 expression sites have not been fully established. We have raised rabbit antisera against an oligopeptide related to the amino acids 652 to 661 of rat NHE2. Western blot analysis of plasma membrane fractions isolated from rat renal cortex showed that affinity-purified anti-NHE2 antibody detected an 85-kDa protein in apical but not in basolateral membranes. The labeling of this 85-kDa protein was specifically blocked by preincubation of the antibody with its monomeric peptide, indicating specific recognition. Indirect immunolabeling was performed on sections of paraformaldehyde-fixed rat kidney embedded in paraffin. Strong staining was seen in the apical membrane of cortical thick ascending limbs, distal convoluted tubules, and connecting tubules. Much weaker apical staining was found in medullary thick ascending limbs of Henle. In the inner medulla, some thin limbs were intensively labeled by the anti-NHE2 antibody. No staining could be detected in any segments of the proximal tubule and collecting duct. PMID- 9729511 TI - Functional activity of epidermal growth factor receptors in autosomal recessive polycystic kidney disease. AB - Evidence from a number of laboratories suggests a potential role for the epidermal growth factor (EGF)-transforming growth factor-alpha-epidermal growth factor receptor (EGF-R) axis in promoting epithelial hyperplasia and cyst formation in autosomal recessive polycystic kidney disease (ARPKD). As previously reported, in the C57BL-6Jcpk/cpk (CPK), BALB/c-bpk/bpk (BPK), and C3H-orpk/orpk (ORPK) murine models of ARPKD, as well as in human ARPKD and human ADPKD, the EGF R is mislocated to the apical surface of cystic collecting tubule (CT) epithelial cells. The present studies demonstrate that cells from cystic and control CTs can be isolated and that these cells maintain their in vivo EGF-R phenotype in vitro. Domain-specific high-affinity ligand binding was assessed by standard Scatchard analysis, and selective ligand stimulation of apical vs. basolateral EGF-R in these cells was followed by measurement of receptor autophosphorylation and determination of cell proliferation. These studies demonstrate that in vitro apically expressed EGF-Rs exhibit high-affinity binding for EGF, autophosphorylate in response to EGF, and transmit a mitogenic signal when stimulated by the appropriate ligand. PMID- 9729512 TI - Neuronal nitric oxide synthase is expressed in principal cell of collecting duct. AB - We used the RT-PCR technique and immunocytochemical methods to determine the expression of endothelial nitric oxide synthase (eNOS) or neuronal nitric oxide synthase (nNOS) in the cortical collecting duct (CCD) in rats on high-K+ diet. The microdissected CCDs of the rat kidney were lysed, and RT-PCR was carried out using rat nNOS and eNOS gene-specific primers. Southern analysis showed the presence of mRNA of nNOS but not eNOS in the CCD. The presence of nNOS in the CCD was further confirmed by light microscopy. We used the polyclonal nNOS antibody in immunocytochemical studies of the isolated CCD. We found that immunoreactivity to nNOS was present in the CCD and heterogeneous with positive and negative immunostaining. We performed the immunocytochemical studies in the split-open CCD and found that the immunoreactivity to nNOS was detected only in principal cells but not in intercalated cells. We conclude that nNOS is expressed in the rat CCD in rats on high-K+ diet. The presence of nNOS in the CCD is heterogeneous and mainly located in principal cells. PMID- 9729513 TI - Dehydration reverses vasopressin antagonist-induced diuresis and aquaporin-2 downregulation in rats. AB - To examine the involvement of vasopressin and dehydration in the regulation of aquaporin-2 (AQP2) expression in rat kidney, we investigated the effects of treatment for 60 h with the specific V2-receptor antagonist OPC-31260 (OPC), alone and in conjunction with dehydration for the last 12 h. Changes in AQP2 protein and mRNA expression in kidney inner medulla were determined by Western and Northern blotting, and AQP2 distribution was analyzed by immunocytochemistry and immunoelectron microscopy. Treatment with OPC increased urine output fourfold, with a reciprocal decrease in urine osmolality. AQP2 expression decreased to 52 +/- 11% of control levels (n = 12, P < 0.05), and AQP2 was found predominantly in intracellular vesicles in collecting duct principal cells. This is consistent with efficient blockade of the vasopressin-induced AQP2 delivery to the plasma membrane and with the observed increased diuresis. Consistent with this, AQP2 mRNA levels were also reduced in response to prolonged OPC treatment (30 +/- 10% of control levels, n = 9). Five days of treatment with furosemide, despite producing even greater polyuria than OPC, was not associated with downregulation of AQP2 levels, demonstrating that AQP2 downregulation is not secondary to increased urine flow rate or loss of medullary hypertonicity. During 12-h thirsting in the continued presence of OPC, urine output dropped dramatically, to levels not significantly different from that seen in (nonthirsted) control animals. In parallel with this, AQP2 levels rose to control levels. Control experiments confirmed continued effective receptor blockade. These results indicate that the V2-receptor antagonist causes a modest decrease in AQP2 expression that is not a consequence of increased urine flow rate or washout of medullary hypertonicity. However, this decrease is much less marked than that seen in some forms of acquired nephrogenic diabetes insipidus. In conjunction with the effects of thirsting, this suggests that modulation of AQP2 expression is mediated partly, but not exclusively, via V2 receptors. PMID- 9729514 TI - Neutral endopeptidase inhibition potentiates the natriuretic actions of adrenomedullin. AB - Adrenomedullin (ADM) is a potent renal vasodilating and natriuretic peptide possessing a six amino acid disulfide ring. Neutral endopeptidase 24.11 (NEP) is localized in greatest abundance in the kidney and cleaves endogenous peptides like atrial natriuretic peptide, which also possesses a disulfide ring. We hypothesized that NEP inhibition potentiates the natriuretic actions of exogenous ADM in anesthetized dogs (n = 6). We therefore investigated renal function in which one kidney received intrarenal infusion of ADM (1 ng . kg-1 . min-1) while the contralateral kidney served as control before and during the systemic infusion of a NEP inhibitor (Candoxatrilat, 8 microg . kg-1 . min-1; Pfizer). In response to ADM, glomerular filtration rate (GFR) in the ADM kidney did not change, whereas renal blood flow, urine flow (UV), and urinary sodium excretion (UNaV) increased from baseline. Proximal and distal fractional reabsorption of sodium decreased in the ADM-infused kidney. In response to systemic NEP inhibition, UNaV and UV increased further in the ADM kidney. Indeed, DeltaUNaV and DeltaUV were markedly greater in the ADM kidney compared with the control kidney. Plasma ADM was unchanged during ADM infusion but increased during NEP inhibition. In conclusion, the present investigation is the first to demonstrate that NEP inhibition potentiates the natriuretic and diuretic responses to intrarenal ADM. This potentiation occurs secondary to a decrease in tubular sodium reabsorption. Lastly, the increase in plasma ADM during systemic NEP inhibition supports the conclusion that ADM is a substrate for NEP. PMID- 9729515 TI - Partially active channels produced by PKA site mutation of the cloned renal K+ channel, ROMK2 (kir1.2). AB - The activity of the cloned renal K+ channel (ROMK2) is dependent on a balance between phosphorylation and dephosphorylation. There are only three protein kinase A (PKA) sites on ROMK2, with the phosphorylated residues being serine-25 (S25), serine-200 (S200), and serine-294 (S294) (Z.-C. Xu, Y. Yang, and S. C. Hebert. J. Biol. Chem. 271: 9313-9319, 1996). We previously mutated these sites from serine to alanine to study the contribution of each site to overall channel function. Here we have studied each of these single PKA site mutants using the single-channel configuration of the patch-clamp technique. Both COOH-terminal mutations at sites S200A and S294A showed a decreased open channel probability (Po), whereas the NH2-terminal mutation at site S25A showed no change in Po compared with wild-type ROMK2. The decrease in Po for the S200A and S294A mutants was caused by the additional presence of a long closed state. In contrast, the occurrence of the S25A channel was approximately 66% less, suggesting fewer active channels at the membrane. The S200A and S294A channels had different kinetics compared with wild-type ROMK2 channels, showing an increased occurrence of sublevels. Similar kinetics were observed when wild-type ROMK2 was excised and exposed to dephosphorylating conditions, indicating that these effects are specifically a property of the partially phosphorylated channel and not due to an unrelated effect of the mutation. PMID- 9729516 TI - Endothelin-1-induced mesangial cell contraction involves activation of protein kinase C-alpha, -delta, and -epsilon. AB - In endothelin-1 (ET-1)-stimulated mesangial cells, to identify the independent roles of calcium and protein kinase C (PKC) causing contraction, the changes in planar surface area in response to ET-1, ionomycin, or phorbol 12-myristate 13 acetate (PMA) were compared. ET-1, PMA, and ionomycin reduced planar area to 49 +/- 3%, 56 +/- 3%, and 78 +/- 2% of basal (means +/- SE, n = 40-50 cells), respectively. ET-1 or ionomycin increased cytosolic calcium from 80 +/- 7 to 220 +/- 30 nM or 97 +/- 10 to 192 +/- 10 nM, respectively. The myosin light chain kinase inhibitor, ML-7, blunted ET-1- but not PMA-stimulated contraction (82 +/- 3% and 48 +/- 6% of time 0, respectively). Cells pretreated with 10 microM chelerythrine for 1 h or PMA for 24 h failed to contract to either ET-1 or PMA. To identify the specific PKC isoform response to ET-1, cytosolic, membrane, and particulate fractions of mesangial cell lysates were immunoblotted with PKC isoform-specific polyclonal antibodies. ET-1 increased membrane PKC-alpha, delta, and -epsilon to 173 +/- 30%, 162 +/- 26%, and 166 +/- 11% of basal (P < 0.05 vs. basal), respectively, and decreased PKC-delta and PKC-epsilon in the cytosol to 56 +/- 11% and 37 +/- 6% of basal, respectively (P < 0.05). ET-1 increased particulate PKC-delta and PKC-epsilon to 172 +/- 15% and 187 +/- 33% of basal (P < 0.05), respectively. PKC-alpha in the cytosol and particulate fractions was not altered by ET-1, but translocation to the nucleus and cell periphery was observed by confocal immunofluorescence imaging. Ionomycin did not change PKC isoform distribution. PKC-zeta was expressed but unaltered by ET-1. Therefore, mesangial cell ET-1-stimulated contraction not only involves a calcium dependent pathway but also includes the activation of one or more PKC-alpha, delta, and -epsilon, but not PKC-zeta. PMID- 9729517 TI - Expression of HKalpha2 protein is increased selectively in renal medulla by chronic hypokalemia. AB - Our laboratory has demonstrated by Northern analysis that chronic hypokalemia increases HKalpha2 (i.e., alpha-subunit of the colonic H+-K+-ATPase) mRNA abundance in the rat. To determine whether the increase in mRNA correlated with an increase in HKalpha2 protein, an antibody was raised against a synthetic peptide derived from amino acids 686-698 of the HKalpha2 sequence. The anti HKalpha2 antibody hybridized to rat distal colon membranes which migrated at approximately 100 kDa (expected mobility of HKalpha2). HKalpha2 protein was not detected in plasma membranes from rat whole kidney or stomach (100 microg) derived from control animals. The antibody was then used to investigate changes in expression of HKalpha2 in renal cortex, renal medulla, and distal colon in two pathophysiological conditions: 1) chronic hypokalemia (LK) and 2) chronic metabolic acidosis (CMA). In LK rats there was a marked, but selective, increase in the abundance of HKalpha2 protein in membranes prepared from renal medulla. Nevertheless, a corresponding increase in HKalpha2 protein abundance was not observed in membranes prepared from the distal colon of LK rats. HKalpha2 protein abundance in CMA was indistinguishable from controls. Moreover, chronic hypokalemia had no effect on expression of alpha1-Na+-K+-ATPase or HKalpha1 in kidney or distal colon under any experimental condition. Therefore, HKalpha2 protein is tissue- and site-specifically upregulated in response to chronic hypokalemia but not by CMA. Furthermore, this regulatory response is localized to the renal medulla. PMID- 9729518 TI - Reflex effects on components of synchronized renal sympathetic nerve activity. AB - The effects of peripheral thermal receptor stimulation (tail in hot water, n = 8, anesthetized) and cardiac baroreceptor stimulation (volume loading, n = 8, conscious) on components of synchronized renal sympathetic nerve activity (RSNA) were examined in rats. The peak height and peak frequency of synchronized RSNA were determined. The renal sympathoexcitatory response to peripheral thermal receptor stimulation was associated with an increase in the peak height. The renal sympathoinhibitory response to cardiac baroreceptor stimulation was associated with a decrease in the peak height. Although heart rate was significantly increased with peripheral thermal receptor stimulation and significantly decreased with cardiac baroreceptor stimulation, peak frequency was unchanged. As peak height reflects the number of active fibers, reflex increases and decreases in synchronized RSNA are mediated by parallel increases and decreases in the number of active renal nerve fibers rather than changes in the centrally based rhythm or peak frequency. The increase in the number of active renal nerve fibers produced by peripheral thermal receptor stimulation reflects the engagement of a unique group of silent renal sympathetic nerve fibers with a characteristic response pattern to stimulation of arterial baroreceptors, peripheral and central chemoreceptors, and peripheral thermal receptors. PMID- 9729519 TI - Urea and NaCl differentially regulate FAK and RAFTK/PYK2 in mIMCD3 renal medullary cells. AB - Two cytosolic tyrosine kinases, focal adhesion kinase (FAK) and the newly described FAK homolog, related adhesion focal tyrosine kinase (RAFTK, also called PYK2 and CAKbeta), have been implicated in signaling to multiple mitogen activated protein kinase (MAPK) pathways. Therefore, the ability of NaCl and urea to activate these kinases was investigated by in vitro kinase assay and anti phosphotyrosine immunoblotting. RAFTK was promptly but only transiently activated by urea (within 1 min; 45%), whereas NaCl activated this kinase at 1, 5, 15, and 30 min of treatment (35-60%). In contrast, FAK exhibited only subtle regulation by the two solutes; however, the time course of induction was distinct for each solute. NaCl activated FAK at 1, 5, and 15 min (25-40%), whereas urea-inducible FAK activation (30%) was not evident until fully 15 min of treatment. At 5 min of treatment with increasing concentrations of solute, both urea and NaCl activated RAFTK in a dose-dependent and comparable fashion, culminating in an approximately twofold activation at 800 mosmol/kgH2O solute. Consistent with these data, solute treatment also enhanced tyrosine phosphorylation of RAFTK. PMID- 9729520 TI - Peptide YY inhibits vasopressin-stimulated chloride secretion in inner medullary collecting duct cells. AB - mIMCD-k2 cells are derived from the inner medullary collecting duct of a mouse and exhibit electrogenic sodium absorption and cAMP- and vasopressin (AVP) stimulated electrogenic chloride secretion [N. L. Kizer, B. Lewis, and B. A. Stanton. Am. J. Physiol. 268 (Renal Fluid Electrolyte Physiol. 37): F347-F355, 1995; and N. L. Kizer, D. Vandorpe, B. Lewis, B. Bunting, J. Russell, and B. A. Stanton. Am. J. Physiol. 268 (Renal Fluid Electrolyte Physiol. 37): F854-F861, 1995]. The purpose of the present study was to determine how peptide YY (PYY) affects electrogenic Na+ and Cl- current in mIMCD-k2 cells. Short-circuit currents (Isc) were measured across monolayers of mIMCD-k2 cells mounted in Ussing-type chambers. PYY did not alter baseline Isc, nor did it alter Isc in chloride-free conditions, indicating no effect on electrogenic sodium transport. Baseline chloride current in these cells is low; therefore, chloride short circuit current (IClsc) was stimulated with AVP (10 nM) added to the basolateral surface and 10 microM amiloride added to the apical surface. Although apical applications of PYY had no effect, basolateral application of PYY caused attenuation of IClsc, with the maximal inhibitory dose (100 nM) causing 52 +/- 1.3% inhibition (IC50 = 0.11 nM). Inhibition by PYY of IClsc is mediated through the Y2 receptor subtype, as PYY-(3-36) was the only PYY analog tested that caused inhibition and was equipotent to PYY. Inhibition by PYY of IClsc was abolished following incubation with pertussis toxin. We also show that PYY inhibits AVP stimulated cAMP accumulation, with a maximal inhibitory dose (100 nM) causing a 38% +/- 6% inhibition (IC50 = 0.16 nM), comparable to inhibition by PYY of IClsc. We conclude that PYY acts through either Gi or Go to inhibit adenylate cyclase activity, leading to a decrease in AVP-stimulated chloride current. PMID- 9729521 TI - Regulation of human mesangial cell collagen expression by transforming growth factor-beta1. AB - Transforming growth factor (TGF)-beta1 has been implicated in glomerular extracellular matrix accumulation. Since the spectrum and mechanism of changes in collagen turnover have not been fully characterized, we evaluated effects of TGF beta1 on collagen expression by human mesangial cells. TGF-beta1 induced increased alpha1(I), alpha1(III), and alpha1(IV) collagen mRNA expression. Greater mRNA expression of matrix metalloproteinase (MMP)-2 was compensated by increased tissue inhibitor of metalloproteinases (TIMP)-2 mRNA. There was no change in TIMP-1 or membrane-type MMP mRNA expression, whereas MMP-1 mRNA decreased. Types I and IV collagen protein accumulated in both the cell layer and medium. Changes in collagen mRNA and protein occurred within 4 and 8 h, respectively. MMP-2 and TIMP-1 and -2 activities showed little change. Cycloheximide markedly decreased collagen detection within 4 h and reversed late, but not early, changes in alpha1(I) collagen mRNA. In this system, increased synthesis may be more significant than degradation for collagen accumulation, but collagen is short-lived in culture. Diverse TGF-beta1 actions on collagen turnover may be either immediate or mediated through synthesis of regulatory molecules. PMID- 9729523 TI - Increased interleukin-6 levels in nasal lavage samples following experimental influenza A virus infection. AB - Interleukin-6 (IL-6) is a pleotropic cytokine implicated in the pathogenesis of local inflammation during viral upper respiratory infections. This study determined if experimental influenza A virus infection causes local IL-6 production. Seventeen healthy, adult subjects were intranasally inoculated, by course drops, with a safety-tested strain of influenza A/Kawasaki/86 (H1N1) virus. Nasal lavage samples were collected, symptoms were recorded, and expelled nasal secretions were weighed once before and then daily for 8 days after the virus inoculation. Lavage samples were submitted for virus culture and were examined for IL-6 and IL-4 by enzyme-linked immunosorbent assay. The IL-6, but not IL-4, levels were significantly increased in the nasal lavage samples of the 12 subjects who shed virus but not in those of the 5 subjects who did not shed virus. Moreover, the elevations in IL-6 levels were related temporally to the development of nasal symptoms and secretions but not to systemic symptoms. These results suggest a role for locally produced IL-6 in the pathogenesis and expressed symptomatology of influenza A virus infection. PMID- 9729522 TI - Cytokines, plasma immune activation markers, and clinically relevant surrogate markers in human immunodeficiency virus infection. PMID- 9729524 TI - Quantifying serum antiplague antibody with a fiber-optic biosensor. AB - The fiber-optic biosensor, originally developed to detect hazardous biological agents such as protein toxins or bacterial cells, has been utilized to quantify the concentration of serum antiplague antibodies. This biosensor has been used to detect and quantify the plague fraction 1 antigen in serum, plasma, and whole blood samples, but its ability to quantify serum antibodies has not been demonstrated. By using a competitive assay, the concentration of serum antiplague antibodies was ascertained in the range of 2 to 15 microgram/ml. By making simple dilutions, concentrations for 11 serum samples whose antiplague antibody concentrations were unknown were determined and were found to be in good agreement with enzyme-linked immunosorbent assay results. The competitive assay method could be used to effectively determine the exposure to plague of animals or humans or could be applied to other diseases, such as hepatitis or AIDS, where the presence of antibodies is used to diagnose infection. PMID- 9729525 TI - Analysis of specific immunoglobulin G subclass antibodies for serological diagnosis of Echinococcosis by a standard enzyme-linked immunosorbent assay. AB - The potential roles of specific antibodies of the different immunoglobulin G (IgG) subclasses in the serological diagnosis of cystic echinococcosis (CE) and alveolar echinococcosis (AE) were investigated by an enzyme-linked immunosorbent assay based on hydatid fluid as antigen. Specific antibodies of subclass 1 were found to be of major importance. In sera collected at the time of diagnosis (i.e., before any therapeutic intervention was initiated) they could be demonstrated in 14 of 15 sera from patients with CE and in all 12 sera from patients with AE. The most discriminatory and the most specific antibodies found in this study belonged to IgG subclass 4. Only one false-positive reaction was observed with 253 sera from healthy volunteers, and no cross-reactions occurred in 80 sera from patients with different parasitic infections. Specific IgG4 antibodies could be demonstrated in 61.0 to 66.7% (CE) or 47.6 to 66.7% (AE) of the cases. Antibody levels of IgG subclass 2 were elevated only moderately, and subclass 3 antibodies were detected in a few cases only. In addition, nonspecific reactions in sera of healthy volunteers or patients with other parasitic infections could partially be attributed to antibodies of subclasses 2 and 3. PMID- 9729526 TI - Analysis of immunoglobulin A antibodies to Helicobacter pylori in serum and gastric juice in relation to mucosal inflammation. AB - Helicobacter pylori is a major etiologic agent in gastroduodenal disorders. In this study, immunoglobulin A (IgA) antibodies to H. pylori antigens were evaluated in serum and gastric juice specimens obtained from patients with gastritis or peptic ulcers by utilizing antibody capture enzyme-linked immunosorbent assays (ACELISAs). Urease alpha subunit (UA), urease beta subunit (UB), the 66-kDa heat shock protein (HSP), and the 25-kDa protein (25K) were used as antigens for the ACELISAs. The antibody titers of the ACELISAs reflect the ratio of H. pylori-specific IgA to total IgA. The ratio is stable, although the antibody concentration fluctuates in gastric juice. By using ACELISAs it was possible to evaluate quantitatively not only serum IgA antibodies but also gastric juice secretory IgA (S-IgA) antibodies. In both serum IgA and gastric juice S-IgA ACELISAs, the titers of antibody to HSP and 25K were remarkably correlated with the histologic grade of gastritis, whereas those to UA and UB were not strongly correlated with histologic grade. Thus, it is useful for estimating the histologic grade of gastritis to quantify serum IgA and gastric juice S-IgA antibodies to HSP and 25K. PMID- 9729527 TI - A sensitive method for quantifying cytomegalic endothelial cells in peripheral blood from cytomegalovirus-infected patients. AB - A sensitive method has been developed for the quantification of cytomegalic endothelial cells (CEC) in peripheral blood (PB) of patients with active cytomegalovirus (CMV) infections. The three subsequent key steps of this method are density centrifugation to enrich endothelial cells (EC) in the mononuclear cell (MNC) fraction, EC-specific staining, and fluorescence-activated cell sorting (FACS) of EC onto adhesion slides. The FACS method was compared with the conventional method of cytocentrifugation of the MNC fraction onto slides, followed by EC-specific staining. The main advantage of the additional steps for the isolation and quantification of CEC in PB by FACS is a 10-times-greater sensitivity than by cytocentrifugation of the MNC fraction alone. The recovery percentages of EC from whole blood were comparable for both methods. Recoveries of EC obtained after FACS were 53% +/- 16.5%, (mean +/- standard deviation), and recoveries of EC obtained after cytocentrifugation of the MNC fraction were 43% +/- 4.3%. In patients with active CMV infection, 5 to 72 CEC were detected by FACS, equivalent to 0.8 to 9.0 CEC/ml of blood. With this method for isolation and quantification, the characterization of CEC in PB of patients with CMV associated clinical symptoms, as well as the quantification of EC in PB of patients with pathophysiological manifestations involving endothelial damage that are different from those caused by CMV infections, can be performed. PMID- 9729528 TI - Detection of human Toxoplasma-specific immunoglobulins A, M, and G with a recombinant Toxoplasma gondii rop2 protein. AB - The Toxoplasma gondii rhoptry protein Rop2 was expressed in Escherichia coli as a fusion protein containing 44 kDa of the 55-kDa mature Rop2, supplied with six histidyl residues at the N-terminal end (Rop2196-561). Humoral response during Toxoplasma infection of humans was analyzed by immunoglobulin G (IgG), IgA, and IgM enzyme-linked immunosorbent assay with Rop2196-561 as the antigen substrate. The analyzed sera were divided according to T. gondii-specific serological tests (IgG, IgA, or IgM indirect immunofluorescence and IgA or IgM immunosorbent agglutination assay) as group A (IgG+ IgA- IgM-; n = 35), group B (IgG+ IgA+ IgM+; n = 21), group C (IgG+ IgA+ IgM-; n = 5), and group D (IgG+ IgA- IgM+; n = 16). Twenty-six T. gondii-seronegative sera from individuals with other infections were also included (group E). Anti-Rop2 IgG antibodies were detected in 82.8% of group A sera and in 97.6% of the sera with acute-phase marker immunoglobulins (groups B, C, and D). The percentage of IgA antibody reactivity against Rop2196-561 was 17.1% in group A, 50% in group D, and 80.8% in groups B and C. The percentage of IgM antibody reactivity was 0% in groups A and C and 62% in groups B and D. Sera from group E failed to show IgA, IgM, or IgG antibody reactivity. Since T. gondii Rop2 elicits a strong humoral response from an early stage of infection, it is suggested that recombinant Rop2196-561 would be suitable for use in diagnostic systems, in combination with other T. gondii antigens, to detect specific IgG, IgA, and IgM antibodies. PMID- 9729529 TI - Age-related changes in blood lymphocyte subsets of Saudi Arabian healthy children. AB - The age-related changes in absolute and percentage values of lymphocyte subsets in the peripheral blood of healthy children of different ages (1 month to 13 years) were studied by flow cytometry. The absolute and percentage values for most lymphocyte subpopulations differed substantially with age. Comparisons among age groups from infants through adults revealed progressive declines in the absolute numbers of leukocytes, total lymphocytes, and T, B, and natural killer (NK) cells. The percentages of T cells increased with age. Within the T lymphocyte population, the CD8(+) subset increased but the CD4(+) subset decreased, resulting in a declining CD4(+)/CD8(+) ratio. The percentage of B cells declined, but that of NK cells remained unchanged. The percentage of HLA DR+ T cells increased over time, but their number changed inconsistently. Our findings confirm and extend earlier reports on age-related changes in lymphocyte subpopulations. These data should be useful in the interpretation of disease related changes, as well as therapy-dependent alterations, in lymphocyte subsets in children of different age groups. PMID- 9729530 TI - Selection of recombinant, library-derived antibody fragments against p24 for human immunodeficiency virus type 1 diagnostics. AB - By application of combinatorial library technology, we generated the first recombinant antibody fragments directed against the major capsid protein p24 of human immunodeficiency virus type 1 (HIV-1). A library of single-chain Fv fragments (scFvs) was constructed by using the antibody variable-region (V) genes of B cells derived from the spleen of a viral lysate-immunized mouse. Antibodies were selected by panning or by enrichment with biotinylated antigen, yielding four different families of antibody fragments. The different types of scFvs were characterized by affinity measurements, by antigen recognition on Western blots, and by pepscan analysis. The epitope of one of the scFvs is located near the residues involved in CypA binding, thereby making it an attractive candidate for therapeutic applications. Comparison of the V gene sequence of this scFV with that of a previously described monoclonal antibody reactive against this immunodominant epitope revealed the usage of the identical combination of VH and Vkappa regions. Thus, this is one of the rare examples in which the original combination in a library-derived antibody fragment was retrieved. After appropriate affinity and format improvements, the best of our recombinant scFvs may form the basis for a sensitive p24 assay as a measure of viral load. In addition, anti-p24 scFvs could be expressed as intracellular antibodies (intrabodies) to aid in the treatment of HIV infections. PMID- 9729531 TI - Human immunodeficiency virus type 1-like DNA sequences and immunoreactive viral particles with unique association with breast cancer. AB - RAK antigens p120, p42, and p25 exhibit molecular and immunological similarity to the proteins encoded by human immunodeficiency virus type 1 (HIV-1) and are expressed by 95% of breast and gynecological cancer cases in women and prostate cancer cases in men. The binding of an epitope-specific anti-HIV-1 gp120 monoclonal antibody (MAb) (amino acids 308 to 322) to cancer RAK antigens has been found to be inhibited by a peptide derived from variable loop V3 of HIV-1. Breast cancer DNAs of 40 patients were PCR amplified with HIV-1 gp41-derived primers, and all of the samples were found to be positive. The DNA fragments amplified in seven blindly selected breast cancer samples were sequenced. The breast cancer DNA sequences showed at least 90% homology to the HIV-1 gene for gp41. Antisense oligonucleotides complementary to the HIV-1-like sequences inhibited reverse transcriptase activity and inhibited the growth of breast cancer cells in vitro. Viral particles detected in breast cancer cell lines were strongly immunogold labeled with the anti-HIV-1 gp120 MAb. The results obtained strongly suggest that the long-postulated breast cancer virus may, in fact, be related to HIV-1. PMID- 9729532 TI - Enzyme immunoassays for serological diagnosis of bovine brucellosis: A trial in Latin America. AB - The results of a field trial conducted in Latin America with two indirect enzyme linked immunosorbent assays (ELISAs) and two competitive ELISAs (CELISAs) for the detection of bovine antibody to Brucella abortus are reported. One of the CELISA formats performed most accurately. The percentage of positive reactions in the CELISA relative to the selected positive rose bengal agglutination test (RBT) and complement fixation test (CFT) results was 97.47%, the percentage of negatives relative to the selected negative RBT and CFT results for unexposed cattle was 98.32%, and the percentage of negatives in cattle vaccinated with B. abortus 19 was 96.51%. The same assay format under Canadian conditions had an actual sensitivity of 100%, a specificity of 99.90% in nonvaccinates, and a specificity of 97.7% in a strain 19-vaccinated population. Overall, the CELISA performed as expected and the results were not dissimilar from the results obtained in the Canadian study. This provided further evidence that this CELISA can in many instances differentiate infected cattle from those that are vaccinated or infected with a cross-reacting organism while still giving very few false positive or false-negative results. PMID- 9729533 TI - Low relative frequencies of CD26(+) CD4(+) cells in long-term nonprogressing human immunodeficiency virus type 1-infected subjects. AB - A broad antibody panel was used for immunophenotyping of human immunodeficiency virus type 1 (HIV-1)-infected patients who were long-term nonprogressors (LTNP). The LTNP were compared with patients in the early phase of infection and patients who had progressed to advanced immunodeficiency. Changes in CD8(+) subset distribution were observed mainly at acquisition of HIV-1 infection, whereas CD4(+) subset changes appeared during progression of HIV-1 infection. The decreasing levels of CD4(+) cells were characterized by an increasing frequency of cells expressing the activation markers HLA-Dr and CD45RO but not the CD28 surface antigen. The LTNP exhibited significant changes compared to HIV-negative patients in almost all markers. Compared to patients in the early phase of infection, the only difference was a relatively lower frequency of CD4(+) cells expressing CD26 among the LTNP. The results show that HIV-1-infected persons who have no signs of immunodeficiency despite many years of infection have an immunophenotypic pattern that is substantially different from that of noninfected persons. Despite the long duration of infection, the LTNP exhibit a pattern similar to that of newly infected persons, with the exception of lower expression of CD26 on CD4(+) cells. PMID- 9729534 TI - A competitive enzyme-linked immunosorbent assay for measuring the levels of serum antibody to Haemophilus influenzae type b. AB - A competitive ELISA method is described for the measurement of total antibodies to the capsular polysaccharide of Haemophilus influenzae type b (HibCPS) in human sera. The competitive method showed an excellent correlation to the radioantigen binding assay (RABA, or Farr assay) and improved correlation of sera with low titers with respect to the more conventional noncompetitive method. Overestimation of samples in the low concentration range was no longer observed with the competitive ELISA method. The free HibCPS competition allowed us to eliminate the day-to-day background variation typical of some sera; thus, only values representing the true anti-HibCPS response were determined. The use of precoated microplates, which could be stored up to 8 months, greatly improved the speed of the procedure. An overall correlation coefficient of 0. 9660 was found when 407 serum samples with a wide variety of anti-HibCPS antibody levels were tested with the competitive ELISA and RABA. The regression line was very close to the ideal line, with a slope of 1.0045 and an intercept of -0.1996. A subset of 96 serum samples representative of all pre- and postimmunization samples was used to compare the competitive ELISA with a previously described ELISA method. The competitive method performed in two laboratories in different countries showed a better correlation with the RABA. The correlation factors were 0.9770 and 0.9816, respectively, while a factor of 0.9547 was found with the previously described noncompetitive procedure, which was better for this method than previously reported (r = 0.917). Therefore, the competitive ELISA is proposed for the assay of anti-HibCPS titers in sera from vaccinated subjects. PMID- 9729535 TI - Endothelial cells expressing an inflammatory phenotype are lysed by superantigen targeted cytotoxic T cells. AB - The objective of this study was to investigate whether the superantigen staphylococcal enterotoxin A (SEA), which binds to HLA class II and T-cell receptor Vbeta chains, can direct cytotoxic T cells to lyse cytokine-stimulated endothelial cells (EC). In addition, we wanted to determine whether SEA-primed cytotoxic T cells could be targeted to EC surface molecules as a means of a novel cancer immunotherapy. Human umbilical vein EC (HUVEC), dermal microvascular EC (HMVEC), or the EC line EA.hy926 stimulated with gamma interferon (IFN-gamma) or tumor necrosis factor alpha (TNF-alpha) displayed upregulated HLA class II and adhesion molecule (CD54 and CD106) expression, respectively. SEA-primed T cells induced a strong cytotoxicity against IFN-gamma- and TNF-alpha-activated EA.hy926 which had been preincubated with SEA. Blocking of CD54 completely abrogated the T cell attack. SEA-D227A, which has a mutated class II binding site, did not promote any cytotoxicity. A strong lysis was observed when a fusion protein consisting of protein A and SEA-D227A was added together with T cells to TNF alpha-induced EA.hy926 and HUVEC precoated with monoclonal antibodies (MAb) directed against HLA class I, CD54, or CD106 molecules. Finally, an scFv antibody fragment reactive with an unknown EC antigen was fused with SEA-D227A. Both EA.hy926 and HMVEC were efficiently lysed by scFv-SEA-D227A-triggered cytotoxic T cells. Taken together, superantigen-activated T-cell-dependent EC killing was induced when EC expressed an inflammatory phenotype. Moreover, specific MAb targeting of the superantigen to surface antigens induced EC lysis. Our data suggest that directed T-cell-mediated lysis of unwanted proliferating EC, such as those in the tumor microvasculature, can be clinically useful. PMID- 9729536 TI - Role of arachidonic acid and its metabolites in the priming of NADPH oxidase in human polymorphonuclear leukocytes by peritoneal dialysis effluent. AB - Peritoneal dialysis effluent (PDE) contains a low-molecular-weight solute that will activate and prime the NADPH oxidase of human neutrophils via a phospholipase A2 (PLA2)-dependent mechanism. Since the products of PLA2 are known to activate and prime the oxidase we have investigated their role in the dialysis effluent-mediated activation and priming of human neutrophils. NADPH oxidase activity of PDE-primed and -unprimed neutrophils was measured by lucigenin enhanced chemiluminescence in the presence of known inhibitors of the arachidonic acid cascade. Incubation of neutrophils with the nonselective PLA2 inhibitor quinacrine (0 to 100 microM) reduced oxidase activity in both primed and unprimed cells. Furthermore, primed cells were more sensitive to the action of quinacrine than were unprimed cells. We were unable to determine the relative roles of secretory PLA2 (sPLA2) and cytosolic PLA2 (cPLA2) since the selective sPLA2 inhibitor scalaradial (0 to 100 microM) inhibited oxidase activity in both groups of cells by similar degrees, while the specific cPLA2 inhibitor AACO-CF3 (0 to 50 microM) failed to affect activity in either group. Inhibition of platelet activating factor (PAF), cycloxygenase, and 5-lipoxygenase-activating protein by hexanolamino-PAF (0 to 25 microM), flurbiprofen (0 to 25 microM), and MK886 (0 to 5 microM), respectively, had no effect upon oxidase activity. However, the direct inhibition of 5-lipoxygenase by caffeic acid or lipoxin A4 resulted in a similar concentration-dependent attenuation of oxidase activity in both primed and unprimed cells. Leukotriene B4 (LTB4) release from primed neutrophils was comparable to that from unprimed cells with the exception of phorbol myristate acetate-stimulated cells, which released fivefold more LTB4 than control. Taken together, these results suggest that it is arachidonic acid per se, and not its metabolites, that is important in priming of the neutrophil NADPH oxidase by dialysis effluent. PMID- 9729537 TI - Diminished adherence and/or ingestion of virulent Mycobacterium tuberculosis by monocyte-derived macrophages from patients with tuberculosis. AB - The interaction between the macrophage and Mycobacterium tuberculosis is mediated by a variety of macrophage membrane-associated proteins. Complement receptors have been implicated in the adherence of M. tuberculosis to macrophages. In the present work, the adherence and/or ingestion of M. tuberculosis H37Rv to human monocyte-derived macrophages (MDM) from patients with tuberculosis (TB) and healthy controls was measured by microscopical examination, [3H]uracil incorporation, and CFU. The adherence and/or ingestion was enhanced by fresh serum and inhibited by heat inactivation, EDTA treatment, and anti-CR1 and anti CR3 antibodies. Comparison of MDM from TB patients and healthy controls showed that the former exhibited a significantly decreased capacity to adhere and/or ingest M. tuberculosis, as determined by the number of CFU and 3H incorporation. The expression of CR1 (CD35) and CR3 (CD11b/CD18) on MDM from TB patients and healthy controls, as determined by flow cytometry, did not show significant differences. These results suggest that the lower ingestion of M. tuberculosis by MDM from TB patients is not due to defects in complement receptors, and therefore, there might be other molecules involved in the adherence and/or ingestion process that render MDM from TB patients ingest less mycobacteria than those from healthy controls. PMID- 9729538 TI - Effects of anticoagulants and temperature on expression of activation markers CD11b and HLA-DR on human leukocytes. AB - A whole-blood model was used to evaluate the effects of temperature and anticoagulant on the expression of activation markers HLA-DR and CD11b on peripheral leukocytes. Venous blood, anticoagulated with either EDTA or heparin, was obtained from six healthy blood donors and 13 hospitalized patients (8 human immunodeficiency virus type 1-seropositive individuals with concurrent pulmonary tuberculosis and 5 patients with pneumonia). A preliminary evaluation was carried out with whole blood from two of the normal donors, and cells were stained immediately for HLA-DR and CD11b markers or stained after incubation at room temperature or 37 degreesC for 18 h with or without the addition of the cytokines gamma interferon (IFN-gamma), granulocyte-macrophage colony-stimulating factor (GM-CSF), IFN-gamma plus GM-CSF, tumor necrosis factor beta, or interleukin-6. Of the cytokines tested, the combination of IFN-gamma and GM-CSF had the most pronounced modulation of marker expression on polymorphonuclear neutrophils (PMN), in particular, HLA-DR expression, which required induction for its detection. These cytokines were therefore used in further evaluations that considered the above-mentioned effects in the presence of disease. Results indicated that the expression of activation markers on PMN and lymphocytes in whole blood are influenced by the temperature of incubation and the choice of anticoagulant and the effects noted were dependent on (i) the particular cell surface marker, (ii) the cell type being studied, and (iii) the presence or absence of disease. It is therefore recommended that ex vivo whole-blood models for evaluating phenotype or immune function be carefully evaluated for the above mentioned effects. PMID- 9729539 TI - Use of highly encapsulated Streptococcus pneumoniae strains in a flow-cytometric assay for assessment of the phagocytic capacity of serotype-specific antibodies. AB - A phagocytosis assay for Streptococcus pneumoniae based on flow cytometry (FACS) with human polymorphonuclear cells and human complement was developed for the study of human vaccination antisera. Human prevaccination sera already contain high levels of C-polysaccharide (C-PS) antibodies, which are not protective in humans but which might give false positive results in a flow-cytometry-based assay. Cultures of S. pneumoniae grown to log phase on three consecutive days, followed by heat inactivation, yielded stable and highly encapsulated strains for serotypes 6A, 6B, 14, 19F, and 23F. As a result, only serotype-specific antibodies were able to facilitate phagocytosis of these strains, whereas no phagocytosis was observed with antibodies against C-PS or pneumococcal surface proteins. No, or weak, phagocytosis was observed with human prevaccination sera, whereas in general, postvaccination antisera facilitated phagocytosis. A highly significant correlation was observed between enzyme-linked immunosorbent assay titers and FACS phagocytosis titers (r = 0.98, P < 0.001) for serotype 23F pneumococci with human vaccination antisera. For all serotypes, interassay variation was below 10%. Major advantages of this assay over the classical killing assay are that (i) limited amounts of sera are required (10 microliter per titration curve), (ii) 600 samples can be processed in one day by one person, and (iii) cells can be fixed and measurement of the samples can be performed up to 1 week later. PMID- 9729540 TI - Serodiagnosis of neosporosis in individual cows and dairy herds: A comparative study of three enzyme-linked immunosorbent assays. AB - The performance of three enzyme-linked immunosorbent assays (ELISA) for detection of antibodies to Neospora caninum in bovine sera was evaluated by using various categories of sera. Two commercial ELISA methods, one based on chemically fixed intact tachyzoites and one based on a sonicate lysate of whole tachyzoites, were compared with an in-house ELISA based on a detergent lysate of whole tachyzoites. A brief description of the development of the latter ELISA is also given. There was good agreement among all three tests with regard to postabortion sera. By using acute-phase abortion sera from cows with confirmed N. caninum-induced and non-N. caninum-induced abortions, satisfactory levels of sensitivity and specificity were calculated for all tests. In addition, similar test results were obtained with postpartum samples from dams and calves. However, considerable differences were found between test results of sequential samples and cross sectional and total-herd samples. Apparently, these discrepancies were due to different sensitivities of the tests for detection of low antibody levels in chronically infected animals. It is suggested that these differences were primarily due to the use of different antigens and different test sample dilutions. It is concluded that all tests are applicable as an additional diagnostic tool in cases of abortion in cattle and for monitoring of congenitally infected calves. For herd screening, the lysate-based ELISAs appear to be more adequate because of their higher sensitivities. PMID- 9729541 TI - rK39 enzyme-linked immunosorbent assay for diagnosis of Leishmania donovani infection. AB - The rK39 enzyme-linked immunosorbent assay (ELISA) was compared with the direct agglutination test (DAT) for Leishmania donovani infection in the Sudan. rK39 ELISA proved more sensitive than DAT in diagnosis of kala-azar (93 and 80%, respectively); both tests may remain positive up to 24 months after treatment. For patients with post-kala-azar dermal leishmaniasis and individuals with subclinical infection, rK39 ELISA performed as well as DAT but could detect infection 6 months earlier in approximately 40% of patients. Conversion in DAT and rK39 ELISA also occurred in leishmanin skin test (LST)-positive individuals, suggesting active parasite replication (rK39 is an amastigote antigen) in these presumably immune individuals. In contrast to DAT, rK39 ELISA also detected infection in randomly selected LST-positive individuals (in four of six) and endemicity (LST-negative) controls (in one of five). rK39 ELISA appears more sensitive than DAT and may prove an important tool in epidemiological studies. PMID- 9729542 TI - Immune response against the exp-1 protein of Plasmodium falciparum results in antibodies that cross-react with human T-cell lymphotropic virus type 1 proteins. AB - To examine the role of the Plasmodium falciparum Exp-1 blood-stage protein in producing antibodies that cross-react with human T-cell lymphotropic virus type I (HTLV-I) proteins, we studied sera from Indonesian volunteers who seroconverted to malaria after transmigrating to an area where malaria is hyperendemic. Samples from Philippine volunteers, that were used in a prior study that examined malaria antibodies that cross-react with HTLV-I proteins, were also used. Eighty-three percent of the Indonesian transmigrants developed antibodies against the malaria Exp-1 protein by 6 months postmigration. Of these malaria seroconverters, 27% developed false-positive HTLV-I enzyme immunoassay (EIA) immunoreactivity, as indicated by indeterminate HTLV-I Western blot banding patterns. Five of the six Philippine samples tested were HTLV-I EIA false positive and Western blot indeterminate. When a recombinant Exp-1 protein was used in blocking experiments, the HTLV-I Western blot immunoreactivity of sera from both groups was either completely eliminated or greatly reduced. No effect on the Western blot immunoreactivity of truly HTLV-I-positive sera was seen. To determine if immunization with the recombinant Exp-1 protein could elicit the production of HTLV-I antibodies, six mice were inoculated with the recombinant protein. Following administration of three 50-microgram doses of the protein, four of the six mice developed antibodies that cross-reacted with HTLV-I proteins on Western blot. These results indicate that the immune response against the malaria Exp-1 protein may result in HTLV-I-cross-reacting antibodies that can lead to false positive EIA and indeterminant Western blotting results. PMID- 9729543 TI - Antigenic variation in Bacteroides forsythus detected by a checkerboard enzyme linked immunosorbent assay. AB - Evidence indicating that multiple serotypes of Bacteroides forsythus participate in rapidly progressing periodontal infections has not been reported previously. Our aim was to develop an assay for detecting subsets of B. forsythus clinical isolates which differ in serogroup membership and subsets of patients with immunoglobulin G (IgG) responses which differ in serogroup recognition. A checkerboard enzyme-linked immunosorbent assay (ELISA) was used to assess variation in the IgG binding profiles of 22 clinical isolates in sera from 28 patients with early-onset rapidly progressive periodontitis. To accommodate the maximum number of isolates and sera in a given assay run, a multiplate assay grid with standard 96-well microtest plates was established. Single dilutions of individual sera were placed in rows crossing columns of isolate-coated wells, and antigen-specific IgG immobilized in the wells was measured as ELISA absorbance. Pooled sera and isolates were assayed in parallel to serve as negative controls for variation in IgG binding profiles. Correlation and hierarchical cluster analysis of the absorbance data matrix showed that the isolates could be sorted into at least four clusters based on variations in their IgG binding profiles across different sera. Furthermore, at least two patient clusters were defined by variations in their serum IgG antigen recognition profiles across different isolates. We conclude that multiple serogroups of B. forsythus exist and that different serogroups are dominant in the antibody response of different patients. The method applied here could be used to serologically classify clinical isolates of other species which evoke a serum antibody response in patients. PMID- 9729545 TI - A serotyping assay for hepatitis C virus in Southeast Asia. AB - A serotyping assay for hepatitis C virus (HCV) was evaluated with samples from Thailand, where the distribution of HCV genotypes was different from that in Western countries where the assay was designed and validated. The sensitivity of the assay was low (58%) for HCV RNA-positive samples compared to that of the genotyping assay (95%, P < 0.01). In addition, only 36% of anti-HCV-positive but HCV RNA-negative samples could be serotyped. The serotypes and genotypes were identical in 96% of the samples that could be typed by both methods. Most of the samples with genotype 6, which was common in Southeast Asia, were nontypeable by this serotyping assay. PMID- 9729546 TI - Neutrophil chemiluminescence during phagocytosis is inhibited by abnormally elevated levels of acetoacetate: implications for diabetic susceptibility to infections. AB - Human neutrophils by a chemiluminescence assay exhibit diminished phagocytic activity in the presence of abnormally high levels of the serum metabolite acetoacetate. These findings, along with our previous evidence demonstrating myeloperoxidase inhibition by acetoacetate, implicate metabolic ketosis in the inhibition of neutrophil microbicidal activity and thus in increased susceptibility to infections. PMID- 9729544 TI - Effects of the nature of adjuvant and site of parenteral immunization on the serum and mucosal immune responses induced by a nasal boost with a vaccine alone. AB - Outbred OF1 mice were immunized subcutaneously with flu vaccine, either in the neck or in the lumbar region (back), in combination with adjuvants inducing either a Th1- or a Th2-type response, referred to as adjuvants A1 and A2, respectively. After two parenteral immunizations, the mice were boosted intranasally with nonadjuvanted vaccine. The serum response was analyzed after each immunization by measuring specific immunoglobulin A (IgA), IgG1, and IgG2a antibody levels, while the local response (same isotypes) was measured in the salivary glands after the mucosal boost by ELISPOTs. We observed that systemic priming at any of the two sites with a Th2 rather than a Th1 adjuvant dramatically enhanced the mucosal IgG1 and IgA responses following a mucosal boost with unadjuvanted vaccine. In addition, as judged by the IgG2a/IgG1 ratios and serum IgA levels, immunization of mice in the back induced a rise in Th2 response compared to neck immunization with adjuvant A1. In contrast, such back immunization with adjuvant A2 reversed the Th1-Th2 balance in favor of the Th1 response compared to neck immunization. Similar differences were observed in mucosal antibody levels according to the site of priming with one given adjuvant; priming in the back with adjuvant A1 increased the mucosal IgA and IgG1 responses compared to neck priming, while the local IgG2a levels were decreased. The reverse was true for adjuvant A2. Back versus neck priming with this latter adjuvant decreased the mucosal IgG1 response, while local IgG2a levels were increased. The different lymphatic drainages of the two sites of parenteral immunization may explain these differences, due to the targeting of particular lymphoid inductive sites. Some of these sites may represent crossroads between systemic and mucosal immunity. PMID- 9729547 TI - Fundamental concepts in statistics: elucidation and illustration. AB - Fundamental concepts in statistics form the cornerstone of scientific inquiry. If we fail to understand fully these fundamental concepts, then the scientific conclusions we reach are more likely to be wrong. This is more than supposition: for 60 years, statisticians have warned that the scientific literature harbors misunderstandings about basic statistical concepts. Original articles published in 1996 by the American Physiological Society's journals fared no better in their handling of basic statistical concepts. In this review, we summarize the two main scientific uses of statistics: hypothesis testing and estimation. Most scientists use statistics solely for hypothesis testing; often, however, estimation is more useful. We also illustrate the concepts of variability and uncertainty, and we demonstrate the essential distinction between statistical significance and scientific importance. An understanding of concepts such as variability, uncertainty, and significance is necessary, but it is not sufficient; we show also that the numerical results of statistical analyses have limitations. PMID- 9729548 TI - Invited editorial on "Irregularities and power law distributions in the breathing pattern in preterm and term infants". PMID- 9729549 TI - Irregularities and power law distributions in the breathing pattern in preterm and term infants. AB - Unlike older children, young infants are prone to develop unstable respiratory patterns, suggesting important differences in their control of breathing. We examined the irregular breathing pattern in infants by measuring the time interval between breaths ("interbreath interval"; IBI) assessed from abdominal movement during 2 h of sleep in 25 preterm infants at a postconceptional age of 40.5 +/- 5.2 (SD) wk and in 14 term healthy infants at a postnatal age of 8.2 +/- 4 wk. In 10 infants we performed longitudinal measurements on two occasions. We developed a threshold algorithm for the detection of a breath so that an IBI included an apneic period and potentially some periods of insufficient tidal breathing excursions (hypopneas). The probability density distribution (P) of IBIs follows a power law, P(IBI) approximately IBI-alpha, with the exponent alpha providing a statistical measurement of the relative risk of insufficient breathing. With maturation, alpha increased from 2.62 +/- 0.4 at 41. 2 +/- 3.6 wk to 3.22 +/- 0.4 at 47.3 +/- 6.4 wk postconceptional age, indicating a decrease in long hypopneas (for paired data P = 0.002). The statistical properties of IBI were well reproduced in a model of the respiratory oscillator on the basis of two hypotheses: 1) tonic neural inputs to the respiratory oscillator are noisy; and 2) the noise explores a critical region where IBI diverges with decreasing tonic inputs. Accordingly, maturation of infant respiratory control can be explained by the tonic inputs moving away from this critical region. We conclude that breathing irregularities in infants can be characterized by alpha, which provides a link between clinically accessible data and the neurophysiology of the respiratory oscillator. PMID- 9729550 TI - Relationship between T-wave amplitude and oxygen pulse in guinea pigs in hyperbaric helium and hydrogen. AB - Diving is known to induce a change in the amplitude of the T wave (ATw) of electrocardiograms, but it is unknown whether this is linked to a change in cardiovascular performance. We analyzed ATw in guinea pigs at 10-60 atm and 25-36 degreesC, breathing 2% O2 in either helium (heliox; n = 10) or hydrogen (hydrox; n = 9) for 1 h at each pressure. Core temperature and electrocardiograms were detected by using implanted radiotelemeters. O2 consumption rate was measured by using gas chromatography. In a previous study (S. R. Kayar and E. C. Parker. J. Appl. Physiol. 82: 988-997, 1997), we analyzed the O2 pulse, i.e., the O2 consumption rate per heart beat, in the same animals. By multivariate regression analysis, we identified variables that were significant to O2 pulse: body surface area, chamber temperature, core temperature, and pressure. In this study, inclusion of ATw made a significantly better model with fewer variables. After normalizing for chamber temperature and pressure, the O2 pulse increased with increasing ATw in heliox (P = 0.001) but with decreasing ATw in hydrox (P < 0.001). Thus ATw is associated with the differences in O2 pulse for animals breathing heliox vs. hydrox. PMID- 9729551 TI - Effect of tryptophan and of glucose on exercise capacity of horses. AB - We hypothesized that central fatigue may have a role in limiting the endurance capacity of horses. Therefore, we tested the effect of infusing tryptophan and/or glucose on endurance time and plasma concentrations of free tryptophan and other substrates thought to affect tryptophan uptake into the brain of seven mares (3-4 yr of age, 353-435 kg) that ran on a treadmill at 50% of maximal O2 consumption to fatigue. With use of a counterbalanced crossover design, the horses were infused with tryptophan (100 mg/kg in saline solution) or a similar volume of saline solution (placebo) before exercise. During exercise, horses received infusions of glucose (2 g/min, 50% wt/vol) or a similar volume of saline. Thus the treatments were 1) tryptophan and glucose (T & G), 2) tryptophan and placebo (T & P), 3) placebo and glucose (P & G), and 4) placebo and placebo (P & P). Mean heart rate, hematocrit, and concentration of plasma total solids before and during exercise were similar for all trials. Mean time to exhaustion was reduced (P < 0.05) for T & P and T & G compared with P & P [86.1 +/- 6.9 and 87.1 +/- 6.8 vs. 102.3 +/- 10.3 (SE) min], whereas endurance for P & G (122.4 +/- 11.9 min) was greater than for all other trials (P < 0.05). Compared with nontryptophan trials, during the tryptophan trials plasma prolactin increased (P < 0.05) nearly threefold before exercise and almost twofold early in exercise. Muscle glycogen concentrations were reduced (P < 0.05) below preexercise values in the P & G and P & P trials only. However, glucose infusions (P & G) did not affect (P > 0.05) concentrations of plasma free fatty acids or ratios of branched-chain amino acids to free tryptophan. In conclusion, tryptophan infusion reduced endurance time, which was consistent with the central fatigue hypothesis. The failure of glucose infusion to alleviate the effects of tryptophan and the absence of significant muscle glycogen reduction in the tryptophan trials suggest that the early onset of fatigue in the tryptophan trials is not due to a lack of readily available substrate. PMID- 9729552 TI - Pulse pressure response to the strain of the valsalva maneuver in humans with preserved systolic function. AB - Arterial pulse pressure response during the strain phase of the Valsalva maneuver has been proposed as a clinical tool for the diagnosis of left heart failure, whereas responses of subjects with preserved systolic function have been poorly documented. We studied the relationship between the aortic pulse amplitude ratio (i.e., minimum/maximum pulse pressure) during the strain phase of the Valsalva maneuver and cardiac hemodynamics at baseline in 20 adults (42 +/- 14 yr) undergoing routine right and left heart catheterization. They were normal subjects (n = 5) and patients with various forms of cardiac diseases (n = 15), and all had a left ventricular ejection fraction >/=40%. High-fidelity pressures were recorded in the right atrium and the left ventricle at baseline and at the aortic root throughout the Valsalva maneuver. Aortic pulse amplitude ratio 1) did not correlate with baseline left ventricular end-diastolic pressure, cardiac index (thermodilution), or left ventricular ejection fraction (cineangiography) and 2) was positively related to total arterial compliance (area method) (r = 0.59) and to basal mean right atrial pressure (r = 0.57) (each P < 0.01). Aortic pulse pressure responses to the strain were not related to heart rate responses during the maneuver. In subjects with preserved systolic function, the aortic pulse amplitude ratio during the strain phase of the Valsalva maneuver relates to baseline total arterial compliance and right heart filling pressures but not to left ventricular function. PMID- 9729553 TI - Nitric oxide and cutaneous active vasodilation during heat stress in humans. AB - Whether nitric oxide (NO) is involved in cutaneous active vasodilation during hyperthermia in humans is unclear. We tested for a role of NO in this process during heat stress (water-perfused suits) in seven healthy subjects. Two forearm sites were instrumented with intradermal microdialysis probes. One site was perfused with the NO synthase inhibitor NG-nitro-L-arginine methyl ester (L-NAME) dissolved in Ringer solution to abolish NO production. The other site was perfused with Ringer solution only. At those sites, skin blood flow (laser Doppler flowmetry) and sweat rate were simultaneously and continuously monitored. Cutaneous vascular conductance, calculated from laser-Doppler flowmetry and mean arterial pressure, was normalized to maximal levels as achieved by perfusion with the NO donor nitroprusside through the microdialysis probes. Under normothermic conditions, L-NAME did not significantly reduce cutaneous vascular conductance. During hyperthermia, with skin temperature held at 38-38.5 degreesC, internal temperature rose from 36.66 +/- 0.10 to 37.34 +/- 0.06 degreesC (P < 0.01). Cutaneous vascular conductance at untreated sites increased from 12 +/- 2 to 44 +/- 5% of maximum, but only rose from 13 +/- 2 to 30 +/- 5% of maximum at L-NAME treated sites (P < 0.05 between sites) during heat stress. L-NAME had no effect on sweat rate (P > 0.05). Thus cutaneous active vasodilation requires functional NO synthase to achieve full expression. PMID- 9729554 TI - Effects of nitric oxide synthase inhibition on cutaneous vasodilation during body heating in humans. AB - We sought to examine further the potential role of nitric oxide (NO) in the neurally mediated cutaneous vasodilation in nonacral skin during body heating in humans. Six subjects were heated with a water-perfused suit while cutaneous blood flow was measured by using laser-Doppler flowmeters placed on both forearms. The NO synthase inhibitor NG-monomethyl-L-arginine (L-NMMA) was given selectively to one forearm via a brachial artery catheter after marked cutaneous vasodilation had been established. During body heating, oral temperature increased by 1.1 +/- 0.1 degreesC while heart rate increased by 30 +/- 6 beats/min. Mean arterial pressure stayed constant at 84 +/- 2 mmHg. In the experimental forearm, cutaneous vascular conductance (CVC; laser-Doppler) decreased to 86 +/- 5% of the peak response to heating (P < 0.05 vs. pre-L-NMMA values) after L-NMMA infusion. In some subjects, L-NMMA caused CVC to fall by approximately 30%; in others, it had little impact on the cutaneous circulation. CVC in the control arm showed a similar increase with heating, then stayed constant while L-NMMA was given to the contralateral side. These results demonstrate that NO contributes modestly, but not consistently, to cutaneous vasodilation during body heating in humans. They also indicate that NO is not the only factor responsible for the dilation. PMID- 9729555 TI - Prolonged exercise increases peripheral plasma ACTH, CRH, and AVP in male athletes. AB - We wished to determine whether the increased ACTH during prolonged exercise was associated with changes in peripheral corticotropin-releasing hormone (CRH) and/or arginine vasopressin (AVP). Six male triathletes were studied during exercise: 1 h at 70% maximal oxygen consumption, followed by progressively increasing work rates until exhaustion. Data obtained during the exercise session were compared with a nonexercise control session. Venous blood was sampled over a 2-h period for cortisol, ACTH, CRH, AVP, renin, glucose, and plasma osmolality. There were significant increases by ANOVA on log-transformed data in plasma cortisol (P = 0.002), ACTH (P < 0.001), CRH (P < 0.001), and AVP (P < 0.03) during exercise compared with the control day. A variable increase in AVP was observed after the period of high-intensity exercise. Plasma osmolality rose with exercise (P < 0.001) and was related to plasma AVP during submaximal exercise (P < 0.03) but not with the inclusion of data that followed the high-intensity exercise. This indicated an additional stimulus to the secretion of AVP. The mechanism by which ACTH secretion occurs during exercise involves both CRH and AVP. We hypothesize that high-intensity exercise favors AVP release and that prolonged duration favors CRH release. PMID- 9729556 TI - Fluorescein-labeled dextran concentration is increased in BAL fluid after ANTU induced edema in rats. AB - Several methodologies have been developed to assess alveolocapillary membrane permeability in acute lung injury. The purpose of this study was to determine the reliability of FITC-dextran compared with radioactive tracers to assess lung permeability alterations. After intraperitoneal administration of alpha naphthylthiourea (ANTU, 50 mg/kg) or DMSO-ANTU vehicle, the animals were euthanized and their lungs were studied in an isolated-lung preparation. FITC dextran or radiolabeled tracers were added to the perfusate. At 2 h the bronchoalveolar lavage (BAL) fluid from the ANTU group showed a significantly greater amount of fluorescence in the supernatant after centrifugation of BAL fluid compared with the DMSO group. Consistent results were observed with the radioactive tracers: there was an increase in extravascular albumin space and extravascular lung water compared with the control group. No cleavage of the FITC from the dextran molecule was evident by chromatography comparing samples recovered from the BAL fluid to the pure FITC-dextran molecule. In conclusion, measurement of FITC-dextran in the supernatant of BAL fluid after intravascular administration is a reliable method of assessing lung permeability changes in vivo and ex vivo. PMID- 9729557 TI - Pulmonary vagal innervation is required to establish adequate alveolar ventilation in the newborn lamb. AB - To investigate the effects of bilateral intrathoracic vagotomy on the establishment of continuous breathing and effective gas exchange at birth, we studied 8 chronically instrumented, unanesthetized, sham-operated and 14 vagotomized newborn lambs after a spontaneous, unassisted vaginal delivery. Fetal lambs were instrumented in utero to record sleep states, diaphragmatic electromyogram, blood pressure, arterial pH, and blood-gas tensions. Six of eight sham-operated lambs established effective gas exchange within 10 min of birth, whereas 12 of 14 vagotomized animals developed respiratory acidosis and hypoxemia (P = 0.008). Breathing frequency in vagotomized newborns was significantly lower during the entire postnatal period compared with sham-operated newborns. Vagotomized subjects also remained hypothermic during the entire postnatal period (P < 0.05). Bronchoalveolar lavage indicated an increased minimum surface tension, whereas lung histology showed perivascular edema and partial atelectasis in the vagotomized group. We conclude that stimulation of breathing and effective gas exchange are critically dependent on intact vagal nerves during the transition from fetal to neonatal life. PMID- 9729558 TI - Heavy snoring with upper airway resistance syndrome may induce intrinsic positive end-expiratory pressure. AB - We studied eight heavy snorers with upper airway resistance syndrome to investigate potential effects of sleep on expiratory airway and lung resistance, intrinsic positive end-expiratory pressure, hyperinflation, and elastic inspiratory work of breathing (WOB). Wakefulness and non-rapid-eye-movement sleep with high- and with low-resistance inspiratory effort (H-RIE and L-RIE, respectively) were compared. No differences in breathing pattern were seen across the three conditions. In contrast, we found increases in expiratory airway and lung resistance during H-RIE compared with L-RIE and wakefulness (56 +/- 24, 16 +/- 4, and 11 +/- 4 cmH2O . 1(-1) . s, respectively), with attendant increases in intrinsic positive end-expiratory pressure (5.4 +/- 1.8, 1.4 +/- 0.5, and 1.3 +/- 1.3 cmH2O, respectively) and elastic WOB (6.1 +/- 2.2, 3.7 +/- 1.2, and 3.4 +/- 0.7 J/min, respectively). The increase in WOB during H-RIE is partly caused by the effects of dynamic pulmonary hyperinflation produced by the increased expiratory resistance. Contrary to the Starling model, a multiple-element compliance model that takes into account the heterogeneity of the pharynx may explain flow limitation during expiration. PMID- 9729559 TI - Zero-stress states of human pulmonary arteries and veins. AB - The zero-stress states of the pulmonary arteries and veins from order 3 to order 9 were determined in six normal human lungs within 15 h postmortem. The zero stress state of each vessel was obtained by cutting the vessel transversely into a series of short rings, then cutting each ring radially, which caused the ring to spring open into a sector. Each sector was characterized by its opening angle. The mean opening angle varied between 92 and 163 degrees in the arterial tree and between 89 and 128 degrees in the venous tree. There was a tendency for opening angles to increase as the sizes of the arteries and veins increased. We computed the residual strains based on the experimental measurements and estimated the residual stresses according to Hooke's law. We found that the inner wall of a vessel at the state in which the internal pressure, external pressure, and longitudinal stress are all zero was under compression and the outer wall was in tension, and that the magnitude of compressive stress was greater than the magnitude of tensile stress. PMID- 9729560 TI - Reductions in cardiac output in hypoxic young pigs: systemic and regional perfusion and oxygen metabolism. AB - We tested the hypotheses that, in hypoxic young pigs, reductions in cardiac output restrict systemic oxygen transport to a greater extent than does hypoxia alone and that compensatory responses to this restriction are more effective in higher than in lower priority vasculatures. To study this, 10- to 14-day-old instrumented awake hypoxic (arterial oxygen tension = 39 Torr) pigs were exposed to reduced venous return by inflation of a right atrial balloon-tipped catheter. Blood flow was measured with radionuclide-labeled microspheres, and oxygen metabolism was determined with arterial and venous oxygen contents from appropriate vessels. Hypoxia resulted in a reduction in oxygen tension; increases in cardiac output and perfusion to brain (72% over baseline), heart, adrenal glands, and liver without reductions to other organs except for the spleen; reductions in systemic and intestinal oxygen delivery; and increases in systemic and intestinal oxygen extraction without changes in systemic, cerebral, or intestinal oxygen uptake. During hypoxia, decreasing venous return was associated with increases in arterial lactic acid concentration and central venous pressure; attenuation of the hypoxia-related increase in cardiac output; sustained increases in brain (72% over baseline) and heart perfusion; reductions in lung (bronchial artery), pancreatic, renal, splenic, and intestinal (-50% below baseline) perfusion; decreases in systemic and gastrointestinal oxygen delivery; sustained increases in systemic and intestinal oxygen extraction; and decreases in intestinal oxygen uptake, without changes in cerebral oxygen metabolism. We conclude that when venous return to the heart is reduced in hypoxic young pigs, the hypoxia-related increase in cardiac output was attenuated and the relative reduction in cardiac output was associated with preserved cerebral oxygen uptake and compromised intestinal oxygen uptake. Regional responses to hypoxia combined with relative reductions in cardiac output differ from that of hypoxia alone, with the greatest effects on lower priority organs such as the gastrointestinal tract. PMID- 9729561 TI - Metabolic and exercise endurance effects of coffee and caffeine ingestion. AB - Caffeine (Caf) ingestion increases plasma epinephrine (Epi) and exercise endurance; these results are frequently transferred to coffee (Cof) consumption. We examined the impact of ingestion of the same dose of Caf in Cof or in water. Nine healthy, fit, young adults performed five trials after ingesting (double blind) either a capsule (Caf or placebo) with water or Cof (decaffeinated Cof, decaffeinated with Caf added, or regular Cof). In all three Caf trials, the Caf dose was 4.45 mg/kg body wt and the volume of liquid was 7.15 ml/kg. After 1 h of rest, the subject ran at 85% of maximal O2 consumption until voluntary exhaustion (approximately 32 min in the placebo and decaffeinated Cof tests). In the three Caf trials, the plasma Caf and paraxanthine concentrations were very similar. After 1 h of rest, the plasma Epi was increased (P < 0.05) by Caf ingestion, but the increase was greater (P < 0.05) with Caf capsules than with Cof. During the exercise there were no differences in Epi among the three Caf trials, and the Epi values were all greater (P < 0.05) than in the other tests. Endurance was only increased (P < 0. 05) in the Caf capsule trial; there were no differences among the other four tests. One cannot extrapolate the effects of Caf to Cof; there must be a component(s) of Cof that moderates the actions of Caf. PMID- 9729563 TI - Apneic threshold for CO2 in the anesthetized rat: fundamental properties under steady-state conditions. AB - Experiments were performed to measure the apneic threshold for CO2 and its fundamental properties in anesthetized rats under steady-state conditions. Breathing was detected from diaphragmatic electromyogram activity. Mechanical hyperventilation resulted in apnea once arterial PCO2 (PaCO2) had fallen far enough. Apnea was not a reflex response to lung inflation because it did not occur immediately, was not prevented by vagotomy, and was reversed by raising PaCO2 without changing mechanical hyperventilation. The apneic threshold was measured by hyperventilating rats mechanically with O2 until apnea had occurred and then raising PaCO2 at constant hyperventilation until breathing reappeared. The mean PaCO2 level of the apneic threshold in 42 rats was 32.8 +/- 0.4 Torr. The level of the threshold did not depend on the volume at which the lungs were inflated. The level of the threshold, under steady-state conditions, was the same when approached from hypocapnia as from eupnea. The level of the threshold could be raised by 9 Torr by chronic elevation of the eupneic PaCO2 level by 18 Torr. PMID- 9729562 TI - Effects of 5-day hypoxia on cardiac adrenergic neurotransmission in rats. AB - Chronic hypoxia induces an overall sympathetic hyperactivation associated with a myocardial beta-receptor desensitization. The mechanisms involved in this desensitization were evaluated in 32 male Wistar rats kept in a hypobaric pressure chamber (PO2 = 40 Torr, atmospheric pressure = 450 Torr) for 5 days. In hypoxic compared with normoxic conditions, plasma norepinephrine (NE) levels were higher (2.1 +/- 0.7 vs. 0.6 +/- 0.2 ng/ml) with no difference in the plasma epinephrine levels (2.2 +/- 0.7 vs. 1.8 +/- 0.3 ng/ml). In hypoxia neuronal NE uptake measured by [3H]NE was decreased by 32% in the right ventricle (RV) and by 35% in the left ventricle (LV), and [3H]mazindol in vitro binding showed a decrease in uptake-1 carrier protein density by 38% in the RV and by 41% in the LV. In vitro binding assays with [3H]CGP-12177 indicate beta-adrenoceptor density reduced by 40% in the RV and by 32% in the LV, and this was due to reduced beta1 subtype fraction (competition binding experiments with practolol). Hypoxia reduced the production of cAMP induced by isoproterenol (36% decrease in the RV and 41% decrease in the LV), 5'-guanylylimododiphosphate (40% decrease in the RV and 42% decrease in the LV), and forskolin (39% decrease in the RV and 41% decrease in the LV) but did not alter the effect of MnCl2 and NaF. Quantitation of inhibitory G-protein alpha-subunit by immunochemical analysis showed a 46% increase in the cardiac-specific isoform Gialpha2 in hypoxic hearts. The present data demonstrate that in rats 5-day hypoxia leads to changes in pre- and postsynaptic myocardial adrenergic function. The myocardial desensitization associated with both a reduction in externalized beta1-adrenoceptor and an increase in inhibitory G-protein subunit may be caused by increased synaptic NE levels due to impaired uptake-1 system. PMID- 9729564 TI - Activity of respiratory pump and upper airway muscles during sleep onset. AB - Ventilation decreases at sleep onset. This change is initiated abruptly at alpha theta electroencephalographic transitions. The aim of this study was to determine the contributions of reduced activity in respiratory pump muscles and upper airway dilator muscles to this change. Surface electromyograms over the diaphragm (Di) and intercostal muscles and fine-wire intramuscular electrodes in genioglossus (GG) and tensor palatini (TP) muscles were recorded in nine healthy young men. It was shown that phasic Di and both phasic and tonic TP activities were lower during theta than during alpha activity. Breath-by-breath analysis of the changes at alpha-theta transitions during the sleep-onset period showed a number of changes. At alpha-theta transitions, phasic activity of Di, intercostal, and GG muscles fell and rose again, and phasic and tonic activities of TP fell and remained at low levels during theta. With a state transition from theta to alpha, the phasic and tonic activities of the Di, GG, and TP increased dramatically. It is now clear that the fall in ventilation that occurs with sleep is related to a fall in activities of both upper airway dilator muscles and respiratory pump muscles. PMID- 9729565 TI - Continuous measurement of tympanic temperature with a new infrared method using an optical fiber. AB - The purpose of this study was to investigate the utility of an infrared tympanic thermometry by using an optical fiber for measuring tympanic temperature (Tty). In the head cooling and facial fanning tests during normothermia, right Tty measured by this method (infrared-Tty) and esophageal temperature (Tes) were not affected by decreased temple and forehead skin temperatures, suggesting that the infrared sensor in this system measured the infrared radiation from the tympanic membrane selectively. Eight male subjects took part in passive-heat-stress and progressive-exercise tests. No significant differences among infrared-Tty, the left Tty measured by thermistor (contact-Tty), and Tes were observed at rest or at the end of each experiment, and there was no significant difference in the increase in these core temperatures from rest to the end. Furthermore, there were no significant differences in the core temperature threshold at the onset of sweating and slope (the relationship of sweating rate vs. infrared-Tty and vs. contact-Tty). These results suggest that this method makes it possible to measure Tty accurately, continuously, and more safely. PMID- 9729566 TI - Muscle coordination in cycling: effect of surface incline and posture. AB - The purpose of the present study was to examine the neuromuscular modifications of cyclists to changes in grade and posture. Eight subjects were tested on a computerized ergometer under three conditions with the same work rate (250 W): pedaling on the level while seated, 8% uphill while seated, and 8% uphill while standing (ST). High-speed video was taken in conjunction with surface electromyography (EMG) of six lower extremity muscles. Results showed that rectus femoris, gluteus maximus (GM), and tibialis anterior had greater EMG magnitude in the ST condition. GM, rectus femoris, and the vastus lateralis demonstrated activity over a greater portion of the crank cycle in the ST condition. The muscle activities of gastrocnemius and biceps femoris did not exhibit profound differences among conditions. Overall, the change of cycling grade alone from 0 to 8% did not induce a significant change in neuromuscular coordination. However, the postural change from seated to ST pedaling at 8% uphill grade was accompanied by increased and/or prolonged muscle activity of hip and knee extensors. The observed EMG activity patterns were discussed with respect to lower extremity joint moments. Monoarticular extensor muscles (GM, vastus lateralis) demonstrated greater modifications in activity patterns with the change in posture compared with their biarticular counterparts. Furthermore, muscle coordination among antagonist pairs of mono- and biarticular muscles was altered in the ST condition; this finding provides support for the notion that muscles within these antagonist pairs have different functions. PMID- 9729567 TI - State-space models of insulin and glucose responses to diets of varying nutrient content in men and women. AB - Discrete-time state-space models were developed to describe contemporaneous responses of plasma insulin and glucose of normal human subjects. Male and female subjects ingested three consecutive identical meals from isocaloric diets classified as high-carbohydrate, high-fat, high-protein, or standard. Distinctly different glucose and insulin responses were measured in men and women. A seven state system of linear equations, three in insulin and four in glucose, was identified and estimated to describe responses in men. A six-state system, three in insulin and three in glucose, describes responses in women. Model simulations at 15-min intervals closely match measured concentrations over a 12-h period. Effects of diet content and meal timing on insulin and glucose concentrations were quantified. Dynamic insulin and glucose responses to isocaloric meals of pure carbohydrate, fat, and protein diets were projected on the basis of models developed from mixed diets. The symmetry of the projections indicates that positive excursions in glucose concentrations associated with carbohydrate intake may be matched with negative excursions associated with fat and protein intake to help manage postmeal glucose excursions. PMID- 9729568 TI - Dopaminergic modulation of respiratory motor output in peripherally chemodenervated goats. AB - We examined the ventilatory effects of exogenous dopamine (DA) and norepinephrine (NE) administration in chloralose-anesthetized, paralyzed, artificially ventilated adult goats before and after carotid body denervation (CBD). Intravenous (iv) DA bolus injections and slow iv infusions caused dose-dependent inhibition of phrenic nerve activity (PNA) in carotid body (CB)-intact animals during normoxia and hyperoxia but not during hypercapnia. NE administration in CB intact goats caused dose-dependent inhibition of PNA of similar magnitude to DA trials. The DA D2-receptor agonists quinelorane and quinpirole inhibited PNA, whereas the DA D1-receptor agonist SKF-81297 had no effect. After CBD, the ventilatory depressant effects of DA persisted, but responses were significantly attenuated compared with CB-intact trials. CBD abolished the inhibitory effect of low-dose NE administration but did not alter ventilatory responses to high-dose NE injection. The peripheral DA D2-receptor antagonist domperidone substantially attenuated the inhibitory effects of DA bolus injections and infusions and reversed the inhibitory ventilatory effect of high-dose DA administration to excitation in some animals. The alpha-adrenoceptor antagonist phentolamine had no effect on DA-induced ventilatory depression. Beta-Adrenoceptor stimulation with isoproterenol produced similar hemodynamic effects to DA administration but had no effect on PNA. We conclude that DA and NE exert both CB-mediated and non-CB mediated inhibitory effects on respiratory motor output in anesthetized goats. The ventilatory depressant effects that persist in peripherally chemodenervated animals are DA D2-receptor mediated, but their exact location remains speculative. PMID- 9729569 TI - Image-analysis-based assessment of the effects of the "Ca2+-jump" technique on sarcomere uniformity. AB - A new image analysis-based technique was used to quantitatively examine the effects of the "Ca2+-jump" activation protocol on the maintenance of fiber quality in skinned rabbit psoas muscle fiber segments. Specifically, contractions in pCa 4.6 were preceded by short-duration "preactivation" soaks in a solution in which EGTA was replaced with the low-Ca2+ buffering capacity analog hexamethylenediamine-N, N, N', N'-tetraacetate, which facilitated rapid Ca2+ equilibration within the fiber segments. Fiber quality was assessed by examining the Fourier spectra of the muscle fiber images before, during, and after activation. Segment lengths were typically below 500 micrometer, thus allowing the majority of the sarcomeres to be visualized in the field of view (x200 and x400 magnification). The preactivation protocol resulted in less deterioration of fiber quality with repetitive activation. In addition, there was also a significant reduction in the time required to reach the 50% level of maximum tension, with no significant change in the maximum tension level. PMID- 9729570 TI - Influence of voluntary exercise on hypothalamic norepinephrine. AB - We combined hypothalamic tissue and plasma determinations of norepinephrine, dihydroxyphenylalanine, and dihydroxyphenylglycol with measurements of abdominal fat in voluntary running rats to examine the relationship among exercise training, hypothalamic and sympathetic nervous function, and body fat stores. The hypothalamic concentrations of norepinephrine, dihydroxyphenylalanine, and dihydroxyphenylglycol were reduced after exercise training (P < 0.01), with the amount of norepinephrine being strongly associated with the plasma norepinephrine (r = 0.58, P < 0.05) and dihydroxyphenylglycol (r = 0.65, P = 0.01) concentrations. Exercise training resulted in a diminution in abdominal fat mass (P < 0.01). A strong relationship existed between fat mass and hypothalamic norepinephrine content (r = 0.83, P < 0.001). The presence of a positive relationship between the arterial and hypothalamic norepinephrine levels provides presumptive evidence of an association between noradrenergic neuronal activity of the hypothalamus and sympathetic nervous function. The observation that abdominal fat mass is linked with norepinephrine in the hypothalamus raises the possibility that alterations in body fat stores provide an afferent signal linking hypothalamic function and the activity of the sympathetic nervous system. PMID- 9729571 TI - Effect of nitric oxide synthase inhibition on hypercapnia-induced hypothermia and hyperventilation. AB - Hypercapnia elicits hypothermia in a number of vertebrates, but the mechanisms involved are not well understood. In the present study, we assessed the participation of the nitric oxide (NO) pathway in hypercapnia-induced hypothermia and hyperventilation by means of NO synthase inhibition by using Nomega-nitro-L arginine (L-NNA). Measurements of ventilation, body temperature, and oxygen consumption were performed in awake unrestrained rats before and after L-NNA injection (intraperitoneally) and L-NNA injection followed by hypercapnia (5% CO2). Control animals received saline injections. L-NNA altered the breathing pattern during the control situation but not during hypercapnia. A significant (P < 0.05) drop in body temperature was measured after both L-NNA (40 mg/kg) and 5% inspired CO2, with a drop in oxygen consumption in the first situation but not in the second. Hypercapnia had no effect on L-NNA-induced hypothermia. The ventilatory response to hypercapnia was not changed by L-NNA, even though L-NNA caused a drop in body temperature. The present data indicate that the two responses elicited by hypercapnia, i.e., hyperventilation and hypothermia, do not share NO as a common mediator. However, the L-arginine-NO pathway participates, although in an unrelated way, in respiratory function and thermoregulation. PMID- 9729572 TI - Cytochrome c promoter activity in soleus and white vastus lateralis muscles in rats. AB - Cytochrome c protein and mRNA are 300 and 100% higher, respectively, in the soleus muscle (predominantly slow-twitch oxidative) than the white vastus lateralis (predominately fast-twitch glycolytic) muscle (W. W. Winder, K. M. Baldwin, and J. O. Holloszy. Eur. J. Biochem. 47: 461-467, 1974; M. M. Lai and F. W. Booth. J. Appl. Physiol. 69: 843-848, 1990). However, the mechanisms controlling these differences in cytochrome c mRNA are largely unknown. The present study employed direct plasmid injection techniques to determine whether the proximal promoter (-726 to +610) of the rat somatic cytochrome c gene was more active in the soleus than in white vastus lateralis muscles in rats. No difference between the soleus and white vastus lateralis muscles for the activities of the -726, -631, -489, -326, -215, -159 and -149 cytochrome c promoters was noted. The results of this study suggest that additional elements (outside of -726 to +610) in the cytochrome c gene may be required, or posttranscriptional regulation may account, for the higher cytochrome c mRNA in the slow-twitch oxidative muscle. PMID- 9729573 TI - Caffeine ingestion and metabolic responses of tetraplegic humans during electrical cycling. AB - Normally, caffeine ingestion results in a wide spectrum of neural and hormonal responses, making it difficult to evaluate which are critical regulatory factors. We examined the responses to caffeine (6 mg/kg) ingestion in a group of spinal cord-injured subjects [7 tetraplegic (C5-7) and 2 paraplegic (T4) subjects] at rest and during functional electrical stimulation of their paralyzed limbs to the point of fatigue. Plasma insulin did not change, caffeine had no effect on plasma epinephrine, and there was a slight increase (P < 0. 05) in norepinephrine after 15 min of exercise. Nevertheless, serum free fatty acids were increased (P < 0.05) after caffeine ingestion after 60 min of rest and throughout the first 15 min of exercise, but the respiratory exchange ratio was not affected. The exercise time was increased (P < 0.05) by 6% or 1.26 +/- 0.57 min. These data suggest that caffeine had direct effects on both the adipose tissue and the active muscle. It is proposed that the ergogenic action of caffeine is occurring, at least in part, by a direct action of the drug on muscle. PMID- 9729574 TI - Stop-flow studies of solute uptake in rat lungs. AB - Stop-flow studies were used to characterize solute uptake in isolated rat lungs. These lungs were perfused at 8 or 34 ml/min for 10-28 s with solutions containing 125I-albumin and two or more of the following diffusible indicators: [3H]mannitol, [14C]urea, 3HOH, 201Tl+, or 86Rb+. After this loading period, flow was stopped for 10-300 s and then resumed to flush out the perfusate that remained in the pulmonary vasculature during the stop interval. Concentrations of 201Tl+ and 86Rb+ in the venous outflow decreased after the stop interval, indicating uptake from exchange vessels during the stop interval. The amount of these K+ analogs lost from the circulation during the stop interval was greater when the intervals were longer. However, losses of 201T1+ at 90 s approached those at 300 s. Because extraction continued after the vasculature had been flushed, vascular levels had presumably fallen to negligible levels during the stop interval. By 90 s of stop flow the vascular volume that was cleared of 201T1+ averaged 0.657 +/- 0.034 (SE) ml in the experiments perfused at 8 ml/min and 0.629 +/- 0.108 ml in those perfused at 34 ml/min. Increases in perfusate K+ decreased the cleared volumes of 201T1+ and 86Rb+. Uptake of [3H]mannitol, [14C]urea, and 3HOH during the stop intervals was observed only when the lungs were loaded at high flow for short intervals. Decreases in 201T1+ and 86Rb+ concentrations in the pulmonary outflow can be used to identify the fraction of the collected samples that were within exchange vessels of the lung during the stop interval and may help determine the distribution of solute and water exchange along the pulmonary vasculature. PMID- 9729575 TI - Vascular resistance and the efficacy of red cell substitutes in a rat hemorrhage model. AB - We have compared polyethylene glycol-modified bovine hemoglobin (PEG-Hb; high O2 affinity, high viscosity, high oncotic pressure) and human hemoglobin cross linked between the alpha-chains (alpha alpha-Hb; low O2 affinity, low viscosity, low oncotic pressure) with a non-O2-carrying plasma expander (pentastarch, high viscosity and oncotic pressure) after a 50% (by volume) exchange transfusion followed by a severe (60% of blood volume) hemorrhage. Mean arterial pressure and systemic vascular resistance rose significantly in the alpha alpha-Hb but not in the PEG-Hb animals. Two-hour survival was greater in the PEG-Hb animals (93%) than in control (35%), pentastarch (8%), or alpha alpha-Hb (6%) animals. In the PEG-Hb animals, there was no disturbance of acid-base balance, significantly less accumulation of lactic acid, and higher cardiac output than in the other groups. The data suggest that the rise in vascular resistance that follows alpha alpha-Hb exchange transfusion offsets the additional O2 transport provided by the cell free hemoglobin. When resistance does not rise, as with PEG-Hb, even relatively small amounts of cell-free hemoglobin appear to be a very effective blood replacement. PMID- 9729576 TI - Epidermal growth factor increases lung liquid clearance in rat lungs. AB - Epidermal growth factor (EGF) has been reported to stimulate the proliferation of epithelial cells and increase Na+ flux and Na+-K+-ATPase function in alveolar epithelial cell monolayers. Increases in Na+-K+-ATPase in alveolar type II cells (AT2) have been associated with increased active Na+ transport and lung edema clearance across the rat alveolar epithelium in a model of proliferative lung injury. Thus we tested whether administration of aerosolized EGF to rat lungs would increase active Na+ transport and lung liquid clearance. Sixteen adult Sprague-Dawley male rats were randomized to three groups. To a group of six rats, an aerosol generated from 20 microgram of EGF in saline was delivered to the lungs, to a second group of five rats only aerosolized saline was delivered, and a third group of five rats without treatment served as the control. Forty-eight hours postaerosolization of rat lungs with EGF there was an approximately 40% increase in active Na+ transport and lung liquid clearance compared with control rats, in the absence of changes in 22Na+, [3H]mannitol, and albumin permeabilities. The Na+-K+-ATPase activity in AT2 cells harvested from these lungs was increased in rats that received aerosolized EGF compared with AT2 cells from both control rats and rats receiving aerosolized saline. These results support the hypothesis that in vivo delivery of EGF aerosols upregulates alveolar epithelial Na+-K+-ATPase and increases lung liquid clearance in rats. PMID- 9729577 TI - Effect of exercise training on passive stiffness in locomotor skeletal muscle: role of extracellular matrix. AB - The purpose of this study was to evaluate the effect of endurance exercise training on both locomotor skeletal muscle collagen characteristics and passive stiffness properties in the young adult and old rat. Young (3-mo-old) and senescent (23-mo-old) male Fischer 344 rats were randomly assigned to either a control or exercise training group [young control (YC), old control (OC), young trained (YT), old trained (OT)]. Exercise training consisted of treadmill running at approximately 70% of maximal oxygen consumption (45 min/day, 5 days/wk, for 10 wk). Passive stiffness (stress/strain) of the soleus (Sol) muscle from all four groups was subsequently measured in vitro at 26 degreesC. Stiffness was significantly greater for Sol muscles in OC rats compared with YC rats, but in OT rats exercise training resulted in muscles with stiffness characteristics not different from those in YC rats. Sol muscle collagen concentration and the level of the nonreducible collagen cross-link hydroxylysylpyridinoline (HP) significantly increased from young adulthood to senescence. Although training had no effect on Sol muscle collagen concentration in either age group, it resulted in a significant reduction in the level of Sol muscle HP in OT rats. In contrast, exercise had no effect on HP in the YT animals. These findings indicate that 10 wk of endurance exercise significantly alter the passive viscoelastic properties of Sol muscle in old but not in young adult rats. The coincidental reduction in the principal collagen cross-link HP also observed in response to training in OT muscle highlights the potential role of collagen in influencing passive muscle viscoelastic properties. PMID- 9729579 TI - Ocular and regional cerebral blood flow in aging Fischer-344 rats. AB - Vascular remodeling and changes in vascular responsiveness occur in the rat cerebrum with old age. This includes reductions in cerebral arteriolar numerical density, cross-sectional area, distensibility, the relative proportion of distensible elements in the cerebral arteriolar wall, and reduced endothelium dependent relaxation. The purpose of this study was to test the hypothesis that old age results in an increase in vascular resistance and, correspondingly, a decrease in blood flow to ocular, regional cerebral, and spinal tissue in the rat. Blood flow was measured in the eye, olfactory bulb, left and right cerebrum, pituitary gland, midbrain, pons, cerebellum, medulla, and spinal cord of juvenile (2-mo-old, n = 6), adult (6-mo-old, n = 7), and aged (24-mo-old, n = 7) male Fischer-344 rats. Arterial pressure and blood flow were used to calculate vascular resistance. Vascular resistance in the eye of aged rats (6.03 +/- 1.08 mmHg . ml-1 . min . 100 g) was higher than that in juvenile (3.83 +/- 0.38 mmHg . ml-1 . min . 100 g) and adult rats (3.12 +/- 0.24 mmHg . ml-1 . min . 100 g). Similarly, resistance in the pons of older rats (2.24 +/- 0.55 mmHg . ml-1 . min . 100 g) was greater than in juvenile (0.66 +/- 0.06 mmHg .ml-1 . min . 100 g) and adult rats (0.80 +/- 0.11 mmHg . ml-1 . min . 100 g). In contrast, vascular resistance in the pituitary gland was lower in the aged rats (juvenile, 3.09 +/- 0.22; adult, 2.79 +/- 0.42; aged, 1.73 +/- 0.32 mmHg . ml-1 . min . 100 g, respectively). Vascular resistance was not different in other cerebral tissues or in the spinal cord in the aged rats. These data suggest that regional cerebral and spinal blood flow and vascular resistance remain largely unchanged in conscious aged rats at rest but that elevations in ocular vascular resistance and, correspondingly, decreases in ocular perfusion with advanced age could have serious adverse effects on visual function. PMID- 9729578 TI - Sarcomere lesion damage occurs mainly in slow fibers of reloaded rat adductor longus muscles. AB - Sarcomere lesions were previously observed with reloading of rat adductor longus muscles after spaceflight and hindlimb unloading (HU). Spaceflown rats displayed more lesioned fibers in the "slow-fiber" region, suggesting a damage-susceptible fiber type. Unloading induces fast myosin expression in some slow fibers, generating hybrid fibers. We examined whether lesion damage differed among slow-, hybrid-, and fast-fiber types in HU-reloaded adductor longus muscles. Temporal HU for 5, 8, 11, 14, and 17 days revealed that hybrid fiber percent, detected by antimyosin immunostaining, peaked at 29 +/- 12% by 14 days. A 14-day HU followed by 12-14 h of voluntary reloading was performed to induce lesions. chi2 analysis showed that slow fibers were preferentially damaged, accounting for 92 +/- 5% of lesioned fibers; hybrid and fast fibers accounted for 7 +/- 4 and <0.5%, respectively. Atrophy did not explain differential lesion damage across fiber types, as slow and hybrid fibers atrophied to a similar extent. Because active myofiber contractions are requisite for lesion formation, selective recruitment of slow fibers most likely explains their damage susceptibility. PMID- 9729580 TI - Blood-gas measurements adjusted for temperature at three sites during incremental exercise in the horse. AB - Rectal temperature (Tre) is often used to adjust measurements of blood gases, but these adjusted measurements may not approximate temperatures during intense exercise at main sites of gas exchange: muscle and lung. To evaluate differences in blood gases between sites, temperatures (T) were measured with thermocouples in the rectum (re), in mixed venous blood (v), in gluteal muscle (mu), and on the skin (sk) in seven Arabian horses as they underwent an incremental exercise test on a treadmill. Blood samples were drawn from the carotid artery and pulmonary artery (mixed venous) 30 s before each increase in speed and during recovery. Blood gases and pH were measured at 37 degreesC, and all variables were adjusted to Tre, Tv, and Tmu. Adjusted variables during exercise and recovery were significantly different from each other at the three sites. Linear and polynomial equations described the time course of venous temperature and from Tre and Tsk during exercise and from Tsk during recovery. Interpretation of changes in muscle metabolism and gas exchanges based on blood-gas measurements is improved if they are adjusted appropriately to Tmu or Tv, which may be predicted from Tsk in addition to Tre during strenuous exercise and from Tsk during recovery. PMID- 9729581 TI - Effects of NaHCO3 loading on acid-base balance, lactate concentration, and performance in racing greyhounds. AB - This investigation examined the effects of NaHCO3 loading on lactate concentration ([La]), acid-base balance, and performance for a 603. 5-m sprint task. Ten greyhounds completed a NaHCO3 (300 mg/kg body weight) and control trial in a crossover design. Results are expressed as means +/- SE. Presprint differences (P < 0.05) were found for NaHCO3 vs. control, respectively, for blood pH (7.47 +/- 0.01 vs. 7.42 +/- 0.01), HCO-3 (28.4 +/- 0.4 vs. 23.5 +/- 0.3 meq/l), and base excess (5.0 +/- 0.3 vs. 0.2 +/- 0.3 meq/l). Peak blood [La] increased (P < 0.05) in NaHCO3 vs. control (20.4 +/- 1.6 vs. 16.9 +/- 1.3 mM, respectively). Relative to control, NaHCO3 produced a greater (P < 0.05) reduction in blood base excess (-18.5 +/- 1.4 vs. -14.1 +/- 0.8 meq/l) and HCO-3 (-17.4 +/- 1.2 vs. -12.8 +/- 0.7 meq/l) from presprint to postexercise. Postexercise peak muscle H+ concentration ([H+]) was higher (P < 0.05) in NaHCO3 vs. control (158.8 +/- 8.8 vs. 137.0 +/- 5.3 nM, respectively). Muscle [H+] recovery half-time (7.2 +/- 1.6 vs. 11.3 +/- 1.6 min) and time to predose values (22.2 +/- 2.4 vs. 32.9 +/- 4.0 min) were reduced (P < 0.05) in NaHCO3 vs. control, respectively. No differences were found in blood [H+] or blood [La] recovery curves or performance times. NaHCO3 increased postexercise blood [La] but did not reduce the muscle or blood acid-base disturbance associated with a 603.5-m sprint or significantly affect performance. PMID- 9729583 TI - Human hydrometry: comparison of multifrequency bioelectrical impedance with 2H2O and bromine dilution. AB - The traditional method of assessing total body water (TBW), extracellular water (ECW), and intracellular water (ICW) has been the use of isotopes, on the basis of the dilution principle. Although the development of bioelectrical impedance techniques has eliminated many of the measurement constraints associated with the dilution methods, the degree of interchangeability between the two methods remains uncertain. We used multifrequency bioelectrical impedance spectroscopy (BIS), 2H2O dilution, and bromine dilution to assess TBW, ECW, and ICW in 469 healthy subjects (248 males, 221 females) aged 3-29 yr. We found that the TBW, ECW, and ICW estimates for the BIS and dilution methods were significantly correlated (r2 = 0.80-0.96, P < 0.0001, SE of the estimate = 2.3-2.7 liters). On the basis of population, the constants used in the BIS analysis could be adjusted so that the mean differences with the dilution methods would become zero. The SD values for the mean differences between the dilution and BIS methods, however, remained significant for both males and females: TBW (+/-2.1-2.8 liters), ECW (+/ 1.4-1.6 liters), and ICW (2.0-3.1 liters). To improve the accuracy of the BIS measurement for an individual within the age range we have examined, further refinement of the constants used in the BIS analysis is needed. PMID- 9729582 TI - Mechanism of leg stiffness adjustment for hopping on surfaces of different stiffnesses. AB - When humans hop in place or run forward, leg stiffness is increased to offset reductions in surface stiffness, allowing the global kinematics and mechanics to remain the same on all surfaces. The purpose of the present study was to determine the mechanism for adjusting leg stiffness. Seven subjects hopped in place on surfaces of different stiffnesses (23-35,000 kN/m) while force platform, kinematic, and electromyographic data were collected. Leg stiffness approximately doubled between the most stiff surface and the least stiff surface. Over the same range of surfaces, ankle torsional stiffness increased 1.75-fold, and the knee became more extended at the time of touchdown (2.81 vs. 2.65 rad). We used a computer simulation to examine the sensitivity of leg stiffness to the observed changes in ankle stiffness and touchdown knee angle. Our model consisted of four segments (foot, shank, thigh, head-arms-trunk) interconnected by three torsional springs (ankle, knee, hip). In the model, an increase in ankle stiffness 1.75 fold caused leg stiffness to increase 1.7-fold. A change in touchdown knee angle as observed in the subjects caused leg stiffness to increase 1.3-fold. Thus both joint stiffness and limb geometry adjustments are important in adjusting leg stiffness to allow similar hopping on different surfaces. PMID- 9729584 TI - Energy requirements and physical activity in free-living older women and men: a doubly labeled water study. AB - Determinants of daily energy needs and physical activity are unknown in free living elderly. This study examined determinants of daily total energy expenditure (TEE) and free-living physical activity in older women (n = 51; age = 67 +/- 6 yr) and men (n = 48; age = 70 +/- 7 yr) by using doubly labeled water and indirect calorimetry. Using multiple-regression analyses, we predicted TEE by using anthropometric, physiological, and physical activity indexes. Data were collected on resting metabolic rate (RMR), body composition, peak oxygen consumption (VO2 peak), leisure time activity, and plasma thyroid hormone. Data adjusted for body composition were not different between older women and men, respectively (in kcal/day): TEE, 2,306 +/- 647 vs. 2,456 +/- 666; RMR, 1,463 +/- 244 vs. 1,378 +/- 249; and physical activity energy expenditure, 612 +/- 570 vs. 832 +/- 581. In a subgroup of 70 women and men, RMR and VO2 peak explained approximately two-thirds of the variance in TEE (R2 = 0.62; standard error of the estimate = +/-348 kcal/day). Crossvalidation of this equation in the remaining 29 women and men was successful, with no difference between predicted and measured TEE (2,364 +/- 398 and 2,406 +/- 571 kcal/day, respectively). The strongest predictors of physical activity energy expenditure (P < 0.05) for women and men were VO2 peak (r = 0.43), fat-free mass (r = 0.39), and body mass (r = 0.34). In summary, RMR and VO2 peak are important independent predictors of energy requirements in the elderly. Furthermore, cardiovascular fitness and fat-free mass are moderate predictors of physical activity in free-living elderly. PMID- 9729585 TI - Regulation of the endogenous NO pathway by prolonged inhaled NO in rats. AB - Nitric oxide (NO) modulates the endogenous NO-cGMP pathway. We determined whether prolonged inhaled NO downregulates the NO-cGMP pathway, which may explain clinically observed rebound pulmonary hypertension. Rats were placed in a normoxic (N; 21% O2) or hypoxic (H; 10% O2) environment with and without inhaled NO (20 parts/million) for 1 or 3 wk. Subsequently, nitric oxide synthase (NOS) and soluble guanylate cyclase (GC) activity and endothelial NOS (eNOS) protein levels were measured. Perfusate cGMP levels and endothelium-dependent and independent vasodilation were determined in isolated lungs. eNOS protein levels and NOS activity were not altered by inhaled NO in N or H rats. GC activity was decreased by 60 +/- 10 and 55 +/- 11% in N and H rats, respectively, after 1 wk of inhaled NO but was not affected after 3 wk. Inhaled NO had no effect on perfusate cGMP in N lungs. Inhaled NO attenuated the increase in cGMP levels caused by 3 wk of H by 57 +/- 11%, but there was no rebound in cGMP after 24 h of recovery. Endothelium-dependent vasodilation was not altered, and endothelium independent vasodilation was not altered (N) or slightly increased (H, 10 +/- 3%) by prolonged inhaled NO. In conclusion, inhaled NO did not alter the endogenous NO-cGMP pathway as determined by eNOS protein levels, NOS activity, or endothelium-dependent vasodilation under N and H conditions. GC activity was decreased after 1 wk; however, GC activity was not altered by 3 wk of inhaled NO and endothelium-independent vasodilation was not decreased. PMID- 9729586 TI - Platelet-activating factor modulates pulmonary vasomotor tone in the perinatal lamb. AB - Eight near-term fetal lambs were studied acutely in utero to determine role of platelet-activating factor (PAF) in the regulation of vasomotor tone in systemic and pulmonary circulations in the immediate perinatal period. Four fetal lambs were studied predelivery and 2 h postdelivery to determine circulating PAF levels. Aortic and pulmonary arterial pressures and cardiac output were measured continuously, and systemic and pulmonary vascular resistances were calculated. Left pulmonary arterial blood flow was also measured in four fetal lambs. After delivery and oxygenation, circulating PAF levels fell significantly. When WEB 2170, a specific PAF-receptor antagonist, was infused to block effect of endogenous PAF in the eight near-term fetal lambs, systemic vascular resistance fell 30% but pulmonary vascular resistance fell dramatically by 68%. Specificity of WEB-2170 was tested in juvenile lambs and was found to be very specific in lowering vasomotor tone only when tone was elevated by action of PAF. Our data show that endogenous PAF levels in the fetus contribute to maintain a high basal systemic and pulmonary vasomotor tone and that a normal fall in circulating PAF levels after birth and oxygenation may facilitate fall in pulmonary vascular resistance at birth. PMID- 9729587 TI - Budesonide affects allergic mucociliary dysfunction. AB - Airway inflammation characterized by neutrophils and free elastase contributes to allergic mucociliary dysfunction. Glucocorticosteroids are the most important anti-inflammatory agents used in the treatment of asthma, but their effect on allergic mucociliary dysfunction is not known. Therefore, we assessed both the prophylactic and therapeutic effects of the glucocorticosteroid budesonide on antigen-induced mucociliary dysfunction in sheep. Tracheal mucus velocity (TMV), a marker of mucociliary clearance, was measured by using a roentgenographic technique. When budesonide was administered either 30 min before or 1 h after airway challenge with Ascaris suum, the antigen-induced fall in TMV at 6 h was prevented. The effects on TMV at 8 and 24 h after challenge were also determined when budesonide and, for comparative purposes, alpha1-protease inhibitor were given 6 h after antigen challenge. Budesonide treatment improved TMV at 8 h, but TMV was not significantly different from antigen alone at 24 h. Treatment with alpha1-protease inhibitor, however, caused only a significant reversal of the antigen-induced fall in TMV at 24 h after challenge; this indicates a more prolonged effect than budesonide. Our results suggest that antiproteases may have a potential role as a therapeutic approach to mucociliary dysfunction in asthma and provide evidence for another means by which glucocorticosteroids contribute to the control of the disease. PMID- 9729588 TI - O2 extraction maintains O2 uptake during submaximal exercise with beta-adrenergic blockade at 4,300 m. AB - Whole body O2 uptake (VO2) during maximal and submaximal exercise has been shown to be preserved in the setting of beta-adrenergic blockade at high altitude, despite marked reductions in heart rate during exercise. An increase in stroke volume at high altitude has been suggested as the mechanism that preserves systemic O2 delivery (blood flow x arterial O2 content) and thereby maintains VO2 at sea-level values. To test this hypothesis, we studied the effects of nonselective beta-adrenergic blockade on submaximal exercise performance in 11 normal men (26 +/- 1 yr) at sea level and on arrival and after 21 days at 4,300 m. Six subjects received propranolol (240 mg/day), and five subjects received placebo. At sea level, during submaximal exercise, cardiac output and O2 delivery were significantly lower in propranolol- than in placebo-treated subjects. Increases in stroke volume and O2 extraction were responsible for the maintenance of VO2. At 4,300 m, beta-adrenergic blockade had no significant effect on VO2, ventilation, alveolar PO2, and arterial blood gases during submaximal exercise. Despite increases in stroke volume, cardiac output and thereby O2 delivery were still reduced in propranolol-treated subjects compared with subjects treated with placebo. Further reductions in already low levels of mixed venous O2 saturation were responsible for the maintenance of VO2 on arrival and after 21 days at 4,300 m in propranolol-treated subjects. Despite similar workloads and VO2, propranolol treated subjects exercised at greater perceived intensity than subjects given placebo at 4,300 m. The values for mixed venous O2 saturation during submaximal exercise in propranolol-treated subjects at 4,300 m approached those reported at simulated altitudes >8,000 m. Thus beta-adrenergic blockade at 4,300 m results in significant reduction in O2 delivery during submaximal exercise due to incomplete compensation by stroke volume for the reduction in exercise heart rate. Total body VO2 is maintained at a constant level by an interaction between mixed venous O2 saturation, the arterial O2-carrying capacity, and hemodynamics during exercise with acute and chronic hypoxia. PMID- 9729589 TI - Does resistive loading decrease diaphragmatic contractility before task failure? AB - While sustaining a load that leads to task failure, it is unclear whether diaphragmatic fatigue develops progressively or occurs only at task failure. We hypothesized that incremental loading produces a progressive decrease in diaphragmatic contractility ever before task failure. Ten subjects generated 60% of maximal transdiaphragmatic pressure (Pdimax) for 2 min, 4 min, and until task failure. Before loading, 20 min after each period of loading, and approximately 20 h after the last period of loading, Pdimax, nonpotentiated and potentiated Pdi twitch pressure (Pditw), and the pattern of respiratory muscle recruitment during a CO2 challenge were recorded. Sensation of inspiratory effort at the 4th min of the task-failure protocol was greater than at the same time in the preceding 4 min protocol. Surprisingly, potentiated Pditw and Pdimax were reduced after 2 min of loading and decreased further after 4 min of loading and after task failure; nonpotentiated Pditw was reduced after 4 min of loading and after task failure. The gastric pressure contribution to tidal breathing during a CO2 challenge decreased progressively in relation to duration of the preceding loading period, whereas expiratory muscle recruitment progressively increased. A rest period of approximately 20 h after task failure was not sufficient to normalize these alterations in respiratory muscle recruitment or fatigue-induced changes in diaphragmatic contractility. In conclusion, while sustaining a mechanical load, the diaphragm progressively fatigued, ever before task failure, and when challenged the rib cage-to-diaphragmatic contribution to tidal breathing and recruitment of the expiratory muscles increased pari passu with duration of the preceding loading. PMID- 9729590 TI - Deterioration of cerebral autoregulation during orthostatic stress: insights from the frequency domain. AB - To determine whether dynamic cerebral autoregulation is impaired during orthostatic stress, cerebral blood flow (CBF) velocity in the middle cerebral artery (transcranial Doppler) and mean arterial pressure (MAP; Finapres) were measured continuously in 12 healthy subjects during ramped maximal lower body negative pressure (LBNP) to presyncope. Velocity and pressure were averaged over 6-min periods of stable data at rest and during LBNP to examine steady-state cerebral hemodynamics. Beat-to-beat variability of velocity and pressure were quantified by a "variation index" (oscillatory amplitude/steady-state mean value) and by power spectral analysis. The dynamic relationship between changes in pressure and velocity was evaluated by the estimates of transfer and coherence function. The results of the study were as follows. Steady-state MAP remained relatively constant during LBNP, whereas CBF velocity decreased progressively by 6, 15, and 21% at -30, -40, and -50 mmHg LBNP, respectively (P < 0.05 compared with baseline). At the maximal level of LBNP (30 s before presyncope) MAP decreased by 9.4% in association with a prominent reduction in velocity by 24% (P < 0.05 compared with baseline). The variation index of pressure increased significantly from 3.8 +/- 0.3% at baseline to 4.5 +/- 0. 6% at -50 mmHg LBNP in association with an increase in the variation index of velocity from 6.0 +/- 0.6 to 8.4 +/- 0.7% (P < 0.05). Consistently, the low- (0.07-0.20 Hz) and high frequency (0.20-0.30 Hz) power of variations in pressure and velocity increased significantly at high levels of LBNP (P < 0.05) in association with an increase in transfer function gain (24% at -50 mmHg, P < 0.05). We conclude that the damping effects of autoregulation on variations in CBF velocity are diminished during orthostatic stress in association with substantial falls in steady-state CBF velocity. We suggest that these changes may contribute in part to the development of presyncope. PMID- 9729591 TI - Effects of lung volume on diaphragm EMG signal strength during voluntary contractions. AB - The use of esophageal recordings of the diaphragm electromyogram (EMG) signal strength to evaluate diaphragm activation during voluntary contractions in humans has recently been criticized because of a possible artifact created by changes in lung volume. Therefore, the first aim of this study was to evaluate whether there is an artifactual influence of lung volume on the strength of the diaphragm EMG during voluntary contractions. The second aim was to measure the required changes in activation for changes in lung volume at a given tension, i.e., the volume activation relationship of the diaphragm. Healthy subjects (n = 6) performed contractions of the diaphragm at different transdiaphragmatic pressure (Pdi) targets (range 20-160 cmH2O) while maintaining chest wall configuration constant at different lung volumes. The diaphragm EMG was recorded with a multiple-array esophageal electrode, with control of signal contamination and electrode positioning. The effects of lung volume on the EMG were studied by comparing the crural diaphragm EMG root mean square (RMS), an index of crural diaphragm activation, with an index of global diaphragm activation obtained by normalizing Pdi to the maximum Pdi at the given muscle length (Pdi/Pdimax@L) at the different lung volumes. We observed a direct relationship between RMS and Pdi/Pdimax@L independent of diaphragm length. The volume-activation relationship of the diaphragm was equally affected by changes in lung volume as the volume-Pdi relationship (60% change from functional residual capacity to total lung capacity). We conclude that the RMS of the diaphragm EMG is not artifactually influenced by lung volume and can be used as a reliable index of diaphragm activation. The volume-activation relationship can be used to infer changes in the length-tension relationship of the diaphragm at submaximal activation/contraction levels. PMID- 9729592 TI - Laryngeal-receptor responses to phasic CO2 in anesthetized cats. AB - We compared the effects of CO2 applied continuously and during expiration on laryngeal-receptor activity in paralyzed, artificially ventilated and nonparalyzed, spontaneously breathing cats by using an isolated larynx, artificially ventilated to approximate a normal respiratory cycle. The majority of quiescent negative-pressure and all cold receptors were excited by 5 and 9% CO2 applied both continuously and during expiration. In general, quiescent positive-pressure, tonic negative-pressure, and tonic positive-pressure receptors were inhibited by 5 and 9% CO2 applied continuously and during expiration. There were no significant differences between responses to 5 and 9% CO2 or to continuous and expired CO2 or between paralyzed and nonparalyzed preparations. In conclusion, laryngeal receptors respond to changes in CO2 concentration occurring during a normal respiratory cycle. Because laryngeal-receptor stimulation exerts reflex effects on ventilation and upper airway muscle activity, these results suggest that airway CO2 plays a role in reflex regulation of breathing and upper airway patency. PMID- 9729594 TI - Naloxone does not alter the "regulated" decrease in core temperature during hypoxemia in guinea pigs. AB - In newborns and adults of a number of species, exposure to acute hypoxemia produces a "regulated" decrease in core temperature, the mechanism of which is unknown. The present experiments were carried out in chronically instrumented newborn (5-10 days of age; n = 59) and older (25-30 days of age; n = 61) guinea pigs to test the hypothesis that the endogenous opioids mediate this regulated decrease in core temperature. During an experiment, core temperature, oxygen consumption, and selected ambient temperature were measured in a thermocline (linear temperature gradient of 10-40 degreesC) during a control period of normoxemia, an experimental period of normoxemia or hypoxemia (inspired oxygen fraction 0.10), and during a recovery period of normoxemia following an intraperitoneal injection of naloxone hydrochloride (a nonspecific opioid antagonist; 1, 2, or 4 mg/kg) or vehicle. Naloxone did not significantly alter basal core temperature or the core temperature response to acute hypoxemia in newborn or older guinea pigs. Naloxone did, however, decrease basal oxygen consumption in newborn and older guinea pigs and altered the thermoregulatory effector mechanism used to decrease core temperature during hypoxemia in the newborn guinea pigs. Our data do not support the hypothesis that the endogenous opioids mediate the regulated decrease in core temperature that occurs in newborn and older guinea pigs during exposure to acute hypoxemia. PMID- 9729593 TI - Angiogenic growth factor mRNA responses to passive and contraction-induced hyperperfusion in skeletal muscle. AB - It has been proposed that, in skeletal muscle, the angiogenic response to exercise may be signaled by the increase in muscle blood flow, via biomechanical changes in the microcirculation (increased shear stress and/or wall tension). To examine this hypothesis, we compared the change in abundance of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), and transforming growth factor-beta1 (TGF-beta1) mRNA in skeletal muscles of the canine leg after 1 h of pump-controlled high blood flow alone (passive hyperperfusion; protocol A) and electrical stimulation of the femoral and sciatic nerves producing muscle contraction (protocol B). The increase in leg blood flow (5.4- and 5. 9-fold change from resting values, respectively) was similar in both groups. Passive hyperperfusion alone did not increase message abundance for VEGF (ratio of mRNA to 18S signals after vs. before hyperperfusion, 0.94 +/- 0.08) or bFGF (1.08 +/- 0.05) but slightly increased that of TGF-beta1 (1.14 +/- 0.07; P < 0.03). In contrast, as previously found in the rat, electrical stimulation provoked more than a threefold increase in VEGF mRNA abundance (3.40 +/- 1.45; P < 0.02). However, electrical stimulation produced no significant changes in either bFGF (1.16 +/- 0.13) or TGF-beta1 (1.31 +/- 0.27). These results suggest that the increased muscle blood flow of exercise does not account for the increased abundance of these angiogenic growth factor mRNA levels in response to acute exercise. We speculate that other factors, such as local hypoxia, metabolite concentration changes, or mechanical effects of contraction per se, may be responsible for the effects of exercise. PMID- 9729595 TI - Exercise training in swine promotes growth of arteriolar bed and capillary angiogenesis in heart. AB - Exercise training induces coronary vascular adaptations. The goal of this study was to contrast the effects of training on capillary and arteriolar growth. Minipigs were trained for 1, 3, 8, and 16 wk and compared with controls. Maximal O2 consumption increased continuously throughout the study. Capillary and arteriolar densities and diameters, and proliferation of vascular cells in these vessels, were determined in perfusion-fixed tissue. The arterioles were subdivided into five groups according to diameter: 10-19.9, 20-30, 31-40, 41-70, and 71-120 microgram. The total vascular bed cross-sectional area increased by 37% at 16 wk, mainly because of an increase in the number of the small arterioles and an increase in the diameter of the larger vessels. Capillary density increased at 3 wk and then returned to control levels by 16 wk; concomitantly, the number of arterioles (20-30 microgram) increased at 16 wk. We speculate that the "extra" capillaries observed at 3 wk were the source of the new arterioles. PMID- 9729596 TI - Effects of four different single exercise sessions on lipids, lipoproteins, and lipoprotein lipase. AB - The purpose of this study was to determine the threshold of exercise energy expenditure necessary to change blood lipid and lipoprotein concentrations and lipoprotein lipase activity (LPLA) in healthy, trained men. On different days, 11 men (age, 26.7 +/- 6.1 yr; body fat, 11.0 +/- 1.5%) completed four separate, randomly assigned, submaximal treadmill sessions at 70% maximal O2 consumption. During each session 800, 1,100, 1,300, or 1,500 kcal were expended. Compared with immediately before exercise, high-density lipoprotein cholesterol (HDL-C) concentration was significantly elevated 24 h after exercise (P < 0.05) in the 1,100-, 1,300-, and 1,500-kcal sessions. HDL-C concentration was also elevated (P < 0.05) immediately after and 48 h after exercise in the 1,500-kcal session. Compared with values 24 h before exercise, LPLA was significantly greater (P < 0.05) 24 h after exercise in the 1,100-, 1,300-, and 1,500-kcal sessions and remained elevated 48 h after exercise in the 1,500-kcal session. These data indicate that, in healthy, trained men, 1,100 kcal of energy expenditure are necessary to elicit increased HDL-C concentrations. These HDL-C changes coincided with increased LPLA. PMID- 9729597 TI - Training-induced alterations of carbohydrate metabolism in women: women respond differently from men. AB - We examined the hypothesis that glucose flux was directly related to relative exercise intensity both before and after a 12-wk cycle ergometer training program [5 days/wk, 1-h duration, 75% peak O2 consumption (VO2 peak)] in healthy female subjects (n = 17; age 23.8 +/- 2.0 yr). Two pretraining trials (45 and 65% of VO2 peak) and two posttraining trials [same absolute workload (65% of old VO2 peak) and same relative workload (65% of new VO2 peak)] were performed on nine subjects by using a primed-continuous infusion of [1-13C]- and [6,6-2H]glucose. Eight additional subjects were studied by using [6, 6-2H]glucose. Subjects were studied postabsorption for 90 min of rest and 1 h of cycling exercise. After training, subjects increased VO2 peak by 25.2 +/- 2.4%. Pretraining, the intensity effect on glucose kinetics was evident between 45 and 65% of VO2 peak with rates of appearance (Ra: 4.52 +/- 0.25 vs. 5.53 +/- 0.33 mg . kg-1 . min-1), disappearance (Rd: 4.46 +/- 0.25 vs. 5.54 +/- 0.33 mg . kg-1 . min-1), and oxidation (Rox: 2.45 +/- 0.16 vs. 4.35 +/- 0.26 mg . kg-1 . min-1) of glucose being significantly greater (P 7% of cases. The leak correction improved the accuracy of ventilatory measurements. The monitoring of leaks is helpful for airtight placement of the face mask and for prevention of serious measurement errors caused by leaks. PMID- 9729599 TI - Muscle use during dynamic knee extension: implication for perfusion and metabolism. AB - Dynamic one-legged knee extension (DKE) is commonly used to examine physiological responses to "aerobic" exercise. Muscle blood flow during DKE is often expressed relative to quadriceps femoris muscle mass irrespective of work rate. This is contrary to the notion that increased force is achieved by recruitment in large muscles. The purpose of this study, therefore, was to determine muscle use during DKE. Six subjects had magnetic resonance images taken of their quadriceps femoris before and after 4 min of DKE at 20 and 40 W. Muscle use was determined by shifts in T2. The cross-sectional area of quadriceps femoris that had an elevated T2 was 16 +/- 1% (mean +/- SE) preexercise, and 54 +/- 5 and 94 +/- 4% after 20- and 40 W DKE, respectively. Volume of quadriceps femoris increased 11.4 +/- 0. 2% (P = 0.006), from 2,230 +/- 233 cm3 before exercise to 2,473 +/- 232 cm3 after 40-W DKE. Extrapolation of these data indicates that 1, 301 +/- 111 cm3 of quadriceps femoris were engaged during 20-W DKE compared with 2,292 +/- 154 cm3 during 40-W DKE. By using muscle blood flow data for submaximal DKE at 20 W [P. Andersen and B. Saltin. J. Physiol. (Lond.) 366: 233-249, 1985; and L. B. Rowell, B. Saltin, B. Kiens, and N. J. Christensen. Am. J. Physiol. 251 (Heart Circ. Physiol. 20): H1038-H1044, 1986] and estimating muscle use in those studies from our data (total muscle mass x 0.54), extrapolated blood flow to active muscle (263 and 278 ml . min-1 . 100 g-1, respectively) is comparable to that obtained during peak aerobic DKE when expressed relative to total muscle mass (243 and 250 ml . min-1 . 100 g-1, respectively). These findings indicate that increased power during aerobic DKE is achieved by recruitment. Additionally, they suggest that blood flow to the active quadriceps femoris muscle does not increase with increases in submaximal work rate but instead is maximal to support aerobic metabolism. Thus increases in muscle blood flow are directed to newly recruited muscle, not to increased perfusion of muscle already engaged. PMID- 9729601 TI - Mutational analysis of AREA, a transcriptional activator mediating nitrogen metabolite repression in Aspergillus nidulans and a member of the "streetwise" GATA family of transcription factors. AB - The transcriptional activator AREA is a member of the GATA family of transcription factors and mediates nitrogen metabolite repression in the fungus Aspergillus nidulans. The nutritional versatility of A. nidulans and its amenability to classical and reverse genetic manipulations make the AREA DNA binding domain (DBD) a useful model for analyzing GATA family DBDs, particularly as structures of two AREA-DNA complexes have been determined. The 109 extant mutant forms of the AREA DBD surveyed here constitute one of the highest totals of eukaryotic transcription factor DBD mutants, are discussed in light of the roles of individual residues, and are compared to corresponding mutant sequence changes in other fungal GATA factor DBDs. Other topics include delineation of the DBD using both homology and mutational truncation, use of frameshift reversion to detect regions of tolerance to mutational change, the finding that duplication of the DBD can apparently enhance AREA function, and use of the AREA system to analyze a vertebrate GATA factor DBD. Some major points to emerge from work on the AREA DBD are (i) tolerance to sequence change (with retention of function) is surprisingly great, (ii) mutational changes in a transcription factor can have widely differing, even opposing, effects on expression of different structural genes so that monitoring expression of one or even several structural genes can be insufficient and possibly misleading, and (iii) a mutational change altering local hydrophobic packing and DNA binding target specificity can markedly influence the behavior of mutational changes elsewhere in the DBD. PMID- 9729603 TI - Nutrient uptake by microorganisms according to kinetic parameters from theory as related to cytoarchitecture. AB - The abilities of organisms to sequester substrate are described by the two kinetic constants specific affinity, a degrees, and maximal velocity Vmax. Specific affinity is derived from the frequency of substrate-molecule collisions with permease sites on the cell surface at subsaturating concentrations of substrates. Vmax is derived from the number of permeases and the effective residence time, tau, of the transported molecule on the permease. The results may be analyzed with affinity plots (v/S versus v, where v is the rate of substrate uptake), which extrapolate to the specific affinity and are usually concave up. A third derived parameter, the affinity constant KA, is similar to KM but is compared to the specific affinity rather than Vmax and is defined as the concentration of substrate necessary to reduce the specific affinity by half. It can be determined in the absence of a maximal velocity measurement and is equal to the Michaelis constant for a system with hyperbolic kinetics. Both are taken as a measure of tau, with departure of KM from KA being affected by permease/enzyme ratios. Compilation of kinetic data indicates a 10(8)-fold range in specific affinities and a smaller (10(3)-fold) range in Vmax values. Data suggest that both specific affinities and maximal velocities can be underestimated by protocols which interrupt nutrient flow prior to kinetic analysis. A previously reported inverse relationship between specific affinity and saturation constants was confirmed. Comparisons of affinities with ambient concentrations of substrates indicated that only the largest a degreesS values are compatible with growth in natural systems. PMID- 9729600 TI - Molecular regulation of beta-lactam biosynthesis in filamentous fungi. AB - The most commonly used beta-lactam antibiotics for the therapy of infectious diseases are penicillin and cephalosporin. Penicillin is produced as an end product by some fungi, most notably by Aspergillus (Emericella) nidulans and Penicillium chrysogenum. Cephalosporins are synthesized by both bacteria and fungi, e.g., by the fungus Acremonium chrysogenum (Cephalosporium acremonium). The biosynthetic pathways leading to both secondary metabolites start from the same three amino acid precursors and have the first two enzymatic reactions in common. Penicillin biosynthesis is catalyzed by three enzymes encoded by acvA (pcbAB), ipnA (pcbC), and aatA (penDE). The genes are organized into a cluster. In A. chrysogenum, in addition to acvA and ipnA, a second cluster contains the genes encoding enzymes that catalyze the reactions of the later steps of the cephalosporin pathway (cefEF and cefG). Within the last few years, several studies have indicated that the fungal beta-lactam biosynthesis genes are controlled by a complex regulatory network, e. g., by the ambient pH, carbon source, and amino acids. A comparison with the regulatory mechanisms (regulatory proteins and DNA elements) involved in the regulation of genes of primary metabolism in lower eukaryotes is thus of great interest. This has already led to the elucidation of new regulatory mechanisms. Furthermore, such investigations have contributed to the elucidation of signals leading to the production of beta lactams and their physiological meaning for the producing fungi, and they can be expected to have a major impact on rational strain improvement programs. The knowledge of biosynthesis genes has already been used to produce new compounds. PMID- 9729604 TI - Growth kinetics of suspended microbial cells: from single-substrate-controlled growth to mixed-substrate kinetics. AB - Growth kinetics, i.e., the relationship between specific growth rate and the concentration of a substrate, is one of the basic tools in microbiology. However, despite more than half a century of research, many fundamental questions about the validity and application of growth kinetics as observed in the laboratory to environmental growth conditions are still unanswered. For pure cultures growing with single substrates, enormous inconsistencies exist in the growth kinetic data reported. The low quality of experimental data has so far hampered the comparison and validation of the different growth models proposed, and only recently have data collected from nutrient-controlled chemostat cultures allowed us to compare different kinetic models on a statistical basis. The problems are mainly due to (i) the analytical difficulty in measuring substrates at growth-controlling concentrations and (ii) the fact that during a kinetic experiment, particularly in batch systems, microorganisms alter their kinetic properties because of adaptation to the changing environment. For example, for Escherichia coli growing with glucose, a physiological long-term adaptation results in a change in KS for glucose from some 5 mg liter-1 to ca. 30 microg liter-1. The data suggest that a dilemma exists, namely, that either "intrinsic" KS (under substrate-controlled conditions in chemostat culture) or micromax (under substrate-excess conditions in batch culture) can be measured but both cannot be determined at the same time. The above-described conventional growth kinetics derived from single-substrate controlled laboratory experiments have invariably been used for describing both growth and substrate utilization in ecosystems. However, in nature, microbial cells are exposed to a wide spectrum of potential substrates, many of which they utilize simultaneously (in particular carbon sources). The kinetic data available to date for growth of pure cultures in carbon-controlled continuous culture with defined mixtures of two or more carbon sources (including pollutants) clearly demonstrate that simultaneous utilization results in lowered residual steady state concentrations of all substrates. This should result in a competitive advantage of a cell capable of mixed-substrate growth because it can grow much faster at low substrate concentrations than one would expect from single substrate kinetics. Additionally, the relevance of the kinetic principles obtained from defined culture systems with single, mixed, or multicomponent substrates to the kinetics of pollutant degradation as it occurs in the presence of alternative carbon sources in complex environmental systems is discussed. The presented overview indicates that many of the environmentally relevant apects in growth kinetics are still waiting to be discovered, established, and exploited. PMID- 9729605 TI - Chlorophyll fluorescence analysis of cyanobacterial photosynthesis and acclimation. AB - Cyanobacteria are ecologically important photosynthetic prokaryotes that also serve as popular model organisms for studies of photosynthesis and gene regulation. Both molecular and ecological studies of cyanobacteria benefit from real-time information on photosynthesis and acclimation. Monitoring in vivo chlorophyll fluorescence can provide noninvasive measures of photosynthetic physiology in a wide range of cyanobacteria and cyanolichens and requires only small samples. Cyanobacterial fluorescence patterns are distinct from those of plants, because of key structural and functional properties of cyanobacteria. These include significant fluorescence emission from the light-harvesting phycobiliproteins; large and rapid changes in fluorescence yield (state transitions) which depend on metabolic and environmental conditions; and flexible, overlapping respiratory and photosynthetic electron transport chains. The fluorescence parameters FV/FM, FV'/FM',qp,qN, NPQ, and phiPS II were originally developed to extract information from the fluorescence signals of higher plants. In this review, we consider how the special properties of cyanobacteria can be accommodated and used to extract biologically useful information from cyanobacterial in vivo chlorophyll fluorescence signals. We describe how the pattern of fluorescence yield versus light intensity can be used to predict the acclimated light level for a cyanobacterial population, giving information valuable for both laboratory and field studies of acclimation processes. The size of the change in fluorescence yield during dark-to-light transitions can provide information on respiration and the iron status of the cyanobacteria. Finally, fluorescence parameters can be used to estimate the electron transport rate at the acclimated growth light intensity. PMID- 9729606 TI - Role of PKA in the timing of developmental events in Dictyostelium cells. AB - The cyclic AMP (cAMP)-dependent protein kinase, PKA, is dispensable for growth of Dictyostelium cells but plays a variety of crucial roles in development. The catalytic subunit of PKA is inhibited when associated with its regulatory subunit but is activated when cAMP binds to the regulatory subunit. Deletion of pkaR or overexpression of the gene encoding the catalytic subunit, pkaC, results in constitutive activity. Development is independent of cAMP in strains carrying these genetic alterations and proceeds rapidly to the formation of both spores and stalk cells. However, morphogenesis is aberrant in these mutants. In the wild type, PKA activity functions in a circuit that can spontaneously generate pulses of cAMP necessary for long-range aggregation. It is also essential for transcriptional activation of both prespore and prestalk genes during the slug stage. During culmination, PKA functions in both prespore and prestalk cells to regulate the relative timing of terminal differentiation. A positive feedback loop results in the rapid release of a signal peptide, SDF-2, when prestalk cells are exposed to low levels of SDF-2. The signal transduction pathway that mediates the response to SDF-2 in both prestalk and prespore cells involves the two component system of DhkA and RegA. When the cAMP phosphodiesterase RegA is inhibited, cAMP accumulates and activates PKA, leading to vacuolation of stalk cells and encapsulation of spores. These studies indicate that multiple inputs regulate PKA activity to control the relative timing of differentiations in Dictyostelium. PMID- 9729602 TI - Molecular and biotechnological aspects of microbial proteases. AB - Proteases represent the class of enzymes which occupy a pivotal position with respect to their physiological roles as well as their commercial applications. They perform both degradative and synthetic functions. Since they are physiologically necessary for living organisms, proteases occur ubiquitously in a wide diversity of sources such as plants, animals, and microorganisms. Microbes are an attractive source of proteases owing to the limited space required for their cultivation and their ready susceptibility to genetic manipulation. Proteases are divided into exo- and endopeptidases based on their action at or away from the termini, respectively. They are also classified as serine proteases, aspartic proteases, cysteine proteases, and metalloproteases depending on the nature of the functional group at the active site. Proteases play a critical role in many physiological and pathophysiological processes. Based on their classification, four different types of catalytic mechanisms are operative. Proteases find extensive applications in the food and dairy industries. Alkaline proteases hold a great potential for application in the detergent and leather industries due to the increasing trend to develop environmentally friendly technologies. There is a renaissance of interest in using proteolytic enzymes as targets for developing therapeutic agents. Protease genes from several bacteria, fungi, and viruses have been cloned and sequenced with the prime aims of (i) overproduction of the enzyme by gene amplification, (ii) delineation of the role of the enzyme in pathogenecity, and (iii) alteration in enzyme properties to suit its commercial application. Protein engineering techniques have been exploited to obtain proteases which show unique specificity and/or enhanced stability at high temperature or pH or in the presence of detergents and to understand the structure-function relationships of the enzyme. Protein sequences of acidic, alkaline, and neutral proteases from diverse origins have been analyzed with the aim of studying their evolutionary relationships. Despite the extensive research on several aspects of proteases, there is a paucity of knowledge about the roles that govern the diverse specificity of these enzymes. Deciphering these secrets would enable us to exploit proteases for their applications in biotechnology. PMID- 9729607 TI - Aerobic anoxygenic phototrophic bacteria. AB - The aerobic anoxygenic phototrophic bacteria are a relatively recently discovered bacterial group. Although taxonomically and phylogenetically heterogeneous, these bacteria share the following distinguishing features: the presence of bacteriochlorophyll a incorporated into reaction center and light-harvesting complexes, low levels of the photosynthetic unit in cells, an abundance of carotenoids, a strong inhibition by light of bacteriochlorophyll synthesis, and the inability to grow photosynthetically under anaerobic conditions. Aerobic anoxygenic phototrophic bacteria are classified in two marine (Erythrobacter and Roseobacter) and six freshwater (Acidiphilium, Erythromicrobium, Erythromonas, Porphyrobacter, Roseococcus, and Sandaracinobacter) genera, which phylogenetically belong to the alpha-1, alpha-3, and alpha-4 subclasses of the class Proteobacteria. Despite this phylogenetic information, the evolution and ancestry of their photosynthetic properties are unclear. We discuss several current proposals for the evolutionary origin of aerobic phototrophic bacteria. The closest phylogenetic relatives of aerobic phototrophic bacteria include facultatively anaerobic purple nonsulfur phototrophic bacteria. Since these two bacterial groups share many properties, yet have significant differences, we compare and contrast their physiology, with an emphasis on morphology and photosynthetic and other metabolic processes. PMID- 9729609 TI - Bacillus thuringiensis and its pesticidal crystal proteins. AB - During the past decade the pesticidal bacterium Bacillus thuringiensis has been the subject of intensive research. These efforts have yielded considerable data about the complex relationships between the structure, mechanism of action, and genetics of the organism's pesticidal crystal proteins, and a coherent picture of these relationships is beginning to emerge. Other studies have focused on the ecological role of the B. thuringiensis crystal proteins, their performance in agricultural and other natural settings, and the evolution of resistance mechanisms in target pests. Armed with this knowledge base and with the tools of modern biotechnology, researchers are now reporting promising results in engineering more-useful toxins and formulations, in creating transgenic plants that express pesticidal activity, and in constructing integrated management strategies to insure that these products are utilized with maximum efficiency and benefit. PMID- 9729608 TI - Insertion sequences. AB - Insertion sequences (ISs) constitute an important component of most bacterial genomes. Over 500 individual ISs have been described in the literature to date, and many more are being discovered in the ongoing prokaryotic and eukaryotic genome-sequencing projects. The last 10 years have also seen some striking advances in our understanding of the transposition process itself. Not least of these has been the development of various in vitro transposition systems for both prokaryotic and eukaryotic elements and, for several of these, a detailed understanding of the transposition process at the chemical level. This review presents a general overview of the organization and function of insertion sequences of eubacterial, archaebacterial, and eukaryotic origins with particular emphasis on bacterial elements and on different aspects of the transposition mechanism. It also attempts to provide a framework for classification of these elements by assigning them to various families or groups. A total of 443 members of the collection have been grouped in 17 families based on combinations of the following criteria: (i) similarities in genetic organization (arrangement of open reading frames); (ii) marked identities or similarities in the enzymes which mediate the transposition reactions, the recombinases/transposases (Tpases); (iii) similar features of their ends (terminal IRs); and (iv) fate of the nucleotide sequence of their target sites (generation of a direct target duplication of determined length). A brief description of the mechanism(s) involved in the mobility of individual ISs in each family and of the structure function relationships of the individual Tpases is included where available. PMID- 9729610 TI - Revision of the nomenclature for the Bacillus thuringiensis pesticidal crystal proteins. AB - The crystal proteins of Bacillus thuringiensis have been extensively studied because of their pesticidal properties and their high natural levels of production. The increasingly rapid characterization of new crystal protein genes, triggered by an effort to discover proteins with new pesticidal properties, has resulted in a variety of sequences and activities that no longer fit the original nomenclature system proposed in 1989. Bacillus thuringiensis pesticidal crystal protein (Cry and Cyt) nomenclature was initially based on insecticidal activity for the primary ranking criterion. Many exceptions to this systematic arrangement have become apparent, however, making the nomenclature system inconsistent. Additionally, the original nomenclature, with four activity-based primary ranks for 13 genes, did not anticipate the current 73 holotype sequences that form many more than the original four subgroups. A new nomenclature, based on hierarchical clustering using amino acid sequence identity, is proposed. Roman numerals have been exchanged for Arabic numerals in the primary rank (e.g., Cry1Aa) to better accommodate the large number of expected new sequences. In this proposal, 133 crystal proteins comprising 24 primary ranks are systematically arranged. PMID- 9729612 TI - Linkage map of Escherichia coli K-12, edition 10: the physical map. AB - A physical map, EcoMap10, of the now completely sequenced Escherichia coli chromosome is presented. Calculated genomic positions for the eight restriction enzymes BamHI, HindIII, EcoRI, EcoRV, BglI, KpnI, PstI, and PvuII are depicted. Both sequenced and unsequenced Kohara/Isono miniset clones are aligned to this calculated restriction map. DNA sequence searches identify the precise locations of insertion sequence elements and repetitive extragenic palindrome clusters. EcoGene10, a revised set of genes and functionally uncharacterized open reading frames (ORFs), is also depicted on EcoMap10. The complete set of unnamed ORFs in EcoGene10 are assigned provisional names beginning with the letter "y" by using a systematic nomenclature. PMID- 9729615 TI - Multiple sites of action of neomycin, Mg2+ and spermine on the NMDA receptors of rat hippocampal CA1 pyramidal neurones. AB - 1. The effects of neomycin on NMDA-evoked currents in isolated CA1 hippocampal pyramidal neurones were investigated and single channel activity was examined in outside-out patches taken from cultured hippocampal neurones. The effects of neomycin on two combinations of NMDA receptor subunits (NR1a-NR2A and NR1a-NR2B) expressed in human embryonic kidney (HEK293) cells were also studied. 2. Neomycin (0. 01-1 mM) caused a potentiation of NMDA-activated currents in all neurones examined. No evidence of a voltage-dependent depression was observed in whole cell recordings. 3. In outside-out patch recordings relatively low concentrations (30 and 100 microM) of neomycin caused a voltage-dependent reduction in single channel current amplitude as well as a large increase in the frequency of channel opening. 4. In saturating concentrations of glycine, neomycin enhanced NMDA activated currents and this glycine-independent enhancement was confirmed using recombinant NR1a-NR2B receptors. Neomycin substantially increased the potency of glycine for the receptor by reducing the rate of dissociation of glycine from the receptor. Neomycin demonstrated a glycine-dependent enhancement of currents mediated by the NR1a-NR2A combination of subunits but a paradoxical depression was observed in saturating concentrations of glycine. 5. Neomycin increased the rate of deactivation of glutamate-activated currents consistent with neomycin causing a reduction in the affinity of the receptor for agonist. 6. These results indicate that neomycin has multiple and complex effects on NMDA receptors. PMID- 9729614 TI - An allosteric interaction between the NMDA receptor polyamine and ifenprodil sites in rat cultured cortical neurones. AB - 1. The atypical NR2B subunit-selective NMDA receptor antagonist ifenprodil was originally believed to act as a competitive antagonist at the polyamine binding site of the NMDA receptor. However, a number of studies have suggested that ifenprodil might bind to a distinct site. 2. Using whole-cell voltage clamp recordings, we have studied the interaction of spermine with both ifenprodil and the related NR2B selective antagonist Ro 8-4304 at the NMDA receptor in rat cultured cortical neurones in the presence of saturating concentrations of glycine. 3. Ifenprodil and Ro 8-4304 inhibited steady-state currents evoked by 100 microM NMDA in the absence of spermine with IC50 values of 0.3 and 0.6 microM, respectively. In the presence of 1 and 3 mM spermine, IC50 values for ifenprodil were 1.4 and 1.8 microM and for Ro 8-4304 they were 3. 0 and 7.5 microM, respectively. 4. In the presence of spermine, the on-time constant of receptor blockade by both antagonists was significantly slower than control and the off-time constant of recovery from receptor blockade following removal of Ro 8-4304 was significantly faster. 5. Fast application of spermine during an NMDA steady-state current in the continuous presence of a subsaturating concentration of either antagonist resulted in a biphasic increase in the current, consistent with a fast increase upon spermine binding and a slow increase resultant from dissociation of antagonist due to spermine binding-induced allosteric reduction in receptor antagonist affinity. In agreement with this, at higher, saturating concentrations of antagonist, the slow increase in current amplitude was markedly reduced or absent. 6. These observations are consistent with a non-competitive, allosteric interaction between spermine and the antagonists, such that spermine binding to the NMDA receptor results in a reduction in receptor affinity for the antagonists and vice versa. 7. The effects of Mg2+ on the NMDA-evoked currents and its interaction with ifenprodil were similar to those of spermine, supporting the suggestion that Mg2+ might be the physiological ligand acting at the spermine site mediating glycine-independent stimulation. PMID- 9729613 TI - Non-pore lining amino acid side chains influence anion selectivity of the human CFTR Cl- channel expressed in mammalian cell lines. AB - 1. The effects of individually mutating two adjacent threonine residues in the sixth membrane-spanning region (TM6) of the cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channel on permeation properties were examined using patch clamp recording from mammalian cell lines stably expressing human CFTR. 2. A number of mutations of T338 significantly affected the permeation properties of the channel. Increases and decreases in single channel conductance were observed for different mutants. Anion selectivity was strongly affected, with no two channel variants sharing the same selectivity sequence. Several mutations led to strong inward rectification of the macroscopic current-voltage relationship. The effects of these mutations on permeation properties were correlated with the size of the amino acid side chain substituted, rather than its chemical nature. 3. Most mutations of T339 resulted in a lack of functional channel expression and apparent misprocessing of the protein. One mutant, T339V, was characterized in detail; its permeation properties were significantly altered, although these effects were not as strong as for T338 mutations. 4. These results suggest an important role for T338 in controlling the permeation properties of the CFTR Cl- channel. It is suggested that mutation of this residue alters the interaction between permeating anions and the channel pore via an indirect effect on the orientation of the TM6 helix. PMID- 9729616 TI - Interaction of memantine and amantadine with agonist-unbound NMDA-receptor channels in acutely isolated rat hippocampal neurons. AB - 1. Using whole-cell patch-clamp techniques, the mechanisms of NMDA channel blockade by amino-adamantane derivatives (AADs) memantine (3, 5-dimethyl aminoadamantane, MEM) and amantadine (1-aminoadamantane, AM) have been studied in rat hippocampal neurons acutely isolated by the vibrodissociation method. A rapid concentration-jump technique was used to replace superfusing solutions. 2. The aspartate (Asp)-induced channel opening greatly accelerated but was not a prerequisite for the recovery from the block by MEM: it was able to leave the channel without agonist assistance. The co-agonist (glycine) as well as the competitive NMDA antagonist DL-2-amino-7-phosphonoheptanoic acid (APV), did not affect this recovery. Membrane depolarization accelerated it, strongly suggesting that this process proceeded via the hydrophilic pathway of the channel. 3. A comparison of the kinetics of the recovery from the block by AADs in the presence and absence of the agonist prompted a hypothesis that the blocker trapped in the channel increased the probability of its transition to the open state. 4. Both MEM and AM were able to block NMDA channels not only in the presence but also in the absence of Asp, although in the latter case the effective blocking concentrations were much higher and the rate of the block development was much smaller than in the former case. The extent of the block increased with the duration of the blocker application. Glycine enhanced this block, while APV attenuated it. The MEM-induced blockade of agonist-unbound channels was enhanced by membrane hyperpolarization and weakened by external Mg2+. These findings strongly suggested that the blocker reached its binding sites via the same hydrophilic pathway both in the presence and absence of the agonist. 5. A comparative analysis of the channel unblocking kinetics in the presence of Asp after their blockade with or without the agonist assistance led us to conclude that in the two cases AADs were bound to the same blocking sites in the channel. PMID- 9729617 TI - N- and L-type calcium channel involvement in depolarization-induced suppression of inhibition in rat hippocampal CA1 cells. AB - 1. We investigated depolarization-induced suppression of inhibition (DSI) under whole-cell voltage clamp in CA1 pyramidal neurons of rat hippocampal slices. DSI, a transient reduction in monosynaptic evoked GABAAergic IPSCs lasting for approximately 1 min, was induced by depolarizing the pyramidal cell to -10 or 0 mV for 1 or 2 s. 2. Raising extracellular Ca2+ concentration increased DSI, and varying the DSI-inducing voltage step showed that the voltage dependence of DSI was like that of high-voltage-activated Ca2+ channels. 3. The P- and Q-type Ca2+ channel blocker omega-agatoxin TK (200 nM and 1 microM) and the R- and T-type Ca2+ channel blocker Ni2+ (100 microM) reduced IPSCs without reducing DSI. 4. The specific N-type Ca2+ channel antagonist omega-conotoxin GVIA (250 nM) reduced IPSC amplitudes and almost completely abolished DSI. 5. Blocking L-type Ca2+ channels with nifedipine (10 microM) had no effect on IPSCs or DSI induced by our standard protocol, but reduced DSI induced by the unclamped Na+- and Ca2+ dependent spikes that occurred when 2(triethylamino)-N-(2,6 dimethylphenyl)acetamide (QX-314) was omitted from the recording pipette solution. 6. Although intracellular Ca2+ stores were not measured, DSI was not affected by cyclopiazonic acid (CPA, 20-40 microM), a blocker of Ca2+ uptake into intracellular stores. 7. We conclude that DSI is initiated by Ca2+ influx through N- and, under certain conditions, L-type Ca2+ channels. PMID- 9729619 TI - cGMP inhibits IP3-induced Ca2+ release in intact rat megakaryocytes via cGMP- and cAMP-dependent protein kinases. AB - 1. Inhibition of inositol 1,4,5-trisphosphate (IP3) receptor-mediated Ca2+ release by cGMP was examined in intact rat megakaryocytes, by using a combination of single cell fluorescence microscopy to monitor intracellular free calcium ([Ca2+]i) and flash photolysis of caged second messengers. 2. Sodium nitroprusside (SNP), a nitric oxide (NO) donor, and the hydrolysis-resistant cGMP analogue 8-(4-chlorophenylthio)guanosine 3',5'-cyclic monophosphate (pCPT-cGMP) inhibited Ca2+ release induced by photolysis of caged IP3. Neither of them affected the rate of Ca2+ removal from the cytoplasm following photolysis of caged Ca2+. 3. Photolysis of the caged NO donor 3-morpholinosydnonimine (SIN-1) during agonist-induced [Ca2+]i oscillations inhibited Ca2+ release without affecting the rate of Ca2+ uptake and/or extrusion. 4. We conclude that the inhibition of IP3-induced Ca2+ release is the principal mechanism of NO-cGMP dependent inhibition of [Ca2+]i mobilization. 5. IPG, a specific peptide inhibitor of cGMP-dependent protein kinase (cGMP-PK), blocked the inhibitory effect of pCPT-cGMP, indicating that the inhibition of IP3-induced Ca2+ release by pCPT-cGMP is mediated by cGMP-PK. However, the simultaneous application of both IPG and IP20, a specific peptide inhibitor of cAMP-dependent protein kinase (cAMP-PK), was required to block the inhibitory effect of SNP. These data strongly suggest that NO-cGMP-dependent inhibition of [Ca2+]i mobilization is mediated via the activation of both cGMP-PK and cAMP-PK. PMID- 9729620 TI - Mechanisms underlying phosphate-induced failure of Ca2+ release in single skinned skeletal muscle fibres of the rat. AB - 1. Single mechanically skinned fibres from rat extensor digitorum longus (EDL) muscles were used to investigate the mechanisms underlying inorganic phosphate (Pi) movements between the myoplasm and the sarcoplasmic reticulum (SR). Force transients elicited by caffeine/low Mg2+ application were used to assess the rate of Pi-induced inhibition of SR Ca2+ release and the subsequent recovery of Ca2+ release following removal of myoplasmic Pi. 2. Myoplasmic Pi reduced SR Ca2+ release in a concentration- and time-dependent manner. A 10 s exposure to 10, 20 and 50 mM myoplasmic Pi reduced SR Ca2+ release by 12 +/- 9, 29 +/- 5 and 82 +/- 5 %, respectively. 3. Removal of myoplasmic ATP at the time of Pi exposure significantly increased the rate and extent of SR Ca2+ release inhibition. For example, Ca2+ release was reduced by 86 +/- 6 % (n = 6) after 20 s exposure to 20 mM Pi in the absence of ATP compared with only 47 +/- 5 % (n = 5) in the presence of ATP. 4. The half and full recovery times for SR Ca2+ release following washout of myoplasmic Pi were 35 s and approximately 7 min, respectively. Recovery of Ca2+ release was unaffected by the absence of ATP during washout of Pi but was prevented when fibres were washed in the presence of high myoplasmic Pi (30 mM). Neither the Pi transporter blocker phenylphosphonic acid (PHPA) nor the anion channel blockers anthracene-9-carboxylic acid (9-AC) and 4,4' diisothiocyanatostilbene-2,2'-disulphonic acid (DIDS) affected the rate of recovery of SR Ca2+ release. 5. These results show that Pi entry and exit from the SR occur primarily through a passive pathway that is insensitive to well known anion channel blockers. Pi inhibition of SR Ca2+ release appears to be a complicated phenomenon influenced by the rate of Pi movement across the SR as well as by the rate, extent and species of Ca2+-Pi precipitate formation in the SR lumen. The more rapid inhibitory effect of Pi in the absence of myoplasmic ATP suggests that Pi may inhibit SR Ca2+ release more efficiently during the later stages of fatigue. PMID- 9729621 TI - Differential regulation of potassium currents by FGF-1 and FGF-2 in embryonic Xenopus laevis myocytes. AB - 1. Fibroblast growth factors (FGFs) are involved in the regulation of many aspects of muscle development. This study investigated their role in regulating voltage-dependent K+ currents in differentiating Xenopus laevis myocytes. Both FGF-1 and FGF-2 are expressed by developing muscle cells, so their actions were compared. Experiments were performed on cultured myocytes isolated from stage 15 embryos. 2. Long-term exposure of the embryonic myocytes to FGF-1 downregulated inward rectifier K+ current (IK(IR)) density as well as both sustained and inactivating voltage-dependent outward K+ currents (IK,S and IK,I, respectively) and their densities. In contrast, FGF-2 upregulated these currents, although, because of an increase in capacitance caused by FGF-2, current density did not change with this factor. 3. The regulation of IK(IR) by FGF-1 was prevented by the cytoplasmic tyrosine kinase inhibitor herbimycin A, but that of IK,S and IK,I was unaffected, indicating that FGF-1 achieves its regulatory effects on electrical development via separate signalling pathways. The receptor tyrosine kinase inhibitor genistein in isolation suppressed K+ currents, but this may have occurred through a channel-blocking mechanism. 4. In many cells, IK, S was found to be composed of two components with differing voltage dependencies of activation. The FGFs brought about an alteration in the amount of total IK,S by equal effects on each component. Conversely, herbimycin A increased the proportion of low voltage-activated current without affecting total current amplitude. Therefore, we suggest that a single species of channel whose voltage dependence is shifted by tyrosine phosphorylation generates IK,S. 5. In summary, FGF-1 and FGF-2 exert opposite effects on voltage-dependent K+ currents in embryonic myocytes and, furthermore, FGF-1 achieves its effects on different K+ currents via separate second messenger pathways. PMID- 9729618 TI - L-type Ca2+ channels in inspiratory neurones of mice and their modulation by hypoxia. AB - 1. Whole-cell (ICa) and single Ca2+ channel currents were measured in inspiratory neurones of neonatal mice (4-12 days old). During whole-cell recordings, ICa slowly declined and disappeared within 10-20 min. The run-down was delayed during hypoxia, indicating ICa potentiation. 2. Ca2+ channels were recorded in cell attached patches using pipettes which contained 110 mM Ba2+. L-type Ca2+ channels exhibited a non-ohmic I-V relationship. The slope conductance was 24 pS below and 50 pS above their null potential. The open probability of the channels increased during oxygen depletion, reaching a maximum 2 min after the onset of hypoxia. Restoration of the oxygen supply brought the channel activity back to initial levels. 3. The channel activity was enhanced by 3-30 microM S(-)Bay K 8644, an agonist of L-type Ca2+ channels. The open probability was increased about 3-fold and the activation curve was shifted by 20 mV in the hyperpolarizing direction. In the presence of the agonist, channel open time increased and long openings appeared. Agonist-modulated channels were also potentiated during oxygen depletion. The effect was due to an increase in open time and a decrease in closed time. The channels were inhibited by bath application of nifedipine (10 microM) and nitrendipine (20 microM). 4. Weak bases such as NH4Cl and TMA increased and weak acids such as sodium acetate and propionate decreased activity of the channels, indicating that they are modulated by intracellular pH. Bath application of 1 microM forskolin enhanced the channel activity, whereas 500 microM NaF suppressed it. 5. L-type Ca2+ channels were modulated by an agonist for mGluR1/5 receptors, (S)-3, 5-dihydrophenylglycine (DHPG, 5 microM). In its presence, the hypoxic facilitation of channels was abolished. 6. After blockade of L-type Ca2+ channels, the respiratory response to hypoxia was modified. The transient enhancement of the respiratory rhythm (augmentation) was no longer evident and the secondary depression occurred earlier. 7. We suggest that L-type Ca2+ channels contribute to the early hypoxic response of the respiratory centre. Glutamate release during hypoxia stimulates postsynaptic metabotropic glutamate receptors, which activate the Ca2+ channels. PMID- 9729622 TI - Divalent ion block of inward rectifier current in human capillary endothelial cells and effects on resting membrane potential. AB - 1. Cultured human capillary endothelial cells (HCEC) contain a large inward rectifier current, IK(IR), that can be abolished by removing external K+ or by adding 50 microM Ba2+. 2. We show that IK(IR) is responsible for maintaining the hyperpolarized potential (-60.6 +/- 0.5 mV, n = 83) of HCEC. Blocking IK(IR) with 50 microM Ba2+ shifts the zero current level and depolarizes HCEC by 36.5 +/- 1.3 mV (n = 4). 3. Increasing external Ca2+ concentration ([Ca2+]o) from 0.5 to 7 mM reduces the magnitude of IK(IR) by 36.5 +/- 2.3 % (n = 5) and depolarizes the cells by 10.33 +/- 2.4 mV (n = 3), whereas decreasing [Ca2+]o from 1.8 to 0.5 mM increases the amplitude of IK(IR) by 6.9 +/- 1.9 % (n = 4). The relationship between [Ca2+]o and the percentage block of IK(IR) gives a Kd value of 5.4 +/- 0.6 mM at -120 mV. 4. IK(IR) is also blocked by other divalent ions, with Ba2+ >> Sr2+ > Mg2+ > Mn2+ = Ca2+, and the block of peak current at -120 mV being 85.3 +/ 3.2 % (n = 5) for 50 microM Ba2+, 62.9 +/- 2.2 % (n = 5) for 5 mM Sr2+, 40.7 +/- 2.5 % (n = 9) for 5 mM Mg2+, 33.4 +/- 2.1 % (n = 5) for 5 mM Mn2+ and 32.9 +/- 2.1 % (n = 5) for 5 mM Ca2+. 5. The voltage dependence of Sr2+ block of peak IK(IR) occurred with a Kd value of 1.0 +/- 0.09 mM for -140 mV, 1.9 +/- 0.16 mM for -130 mV, 3.1 +/- 0.28 mM for -120 mV, 4.6 +/- 0.34 mM for -110 mV and 6.4 +/- 0.5 mM for -100 mV (n = 5), with a calculated electrical distance (delta) of 0.44 from the outside. PMID- 9729623 TI - Activation of bovine tracheal chloride channels by amino group-specific reagents. AB - 1. The predominant Cl- channel in bovine tracheal epithelial cells has a conductance of approximately 71 pS and accounts for more than 80 % of the total chloride conductance. We examined the effects of protein-modifying reagents on channel function and found that amino groups are critically involved in gating. 2. Patch clamp studies showed that lysine-specific reagents, such as dimethyl adipimidate (DMA), significantly increased the channel open probability, but not its conductance. This suggests that modified residues are involved in the gating mechanism, but are distant from the channel permeation pathway. 3. Kinetic analysis of channel activity showed that histograms of open and closed durations could be well fitted by double exponential distributions, suggesting that the channel has at least two open and two closed states. DMA did not change the number of open or closed states, but increased channel mean open time. 4. Since membrane impermeant reagents were effective only from the extracellular side, we conclude that lysine residues in the extracellular domain of the channel are critically involved in gating. These residues may present an important target for site-directed mutagenesis and pharmacological activation of Cl- channels in epithelial cells. PMID- 9729624 TI - Evidence for an electrogenic Na+-HCO3- symport in rat cardiac myocytes. AB - 1. The perforated whole-cell configuration of patch clamp and the pH fluorescent indicator SNARF were used to determine the electrogenicity of the Na+-HCO3- cotransport in isolated rat ventricular myocytes. 2. Switching from Hepes buffer to HCO3- buffer at constant extracellular pH (pHo) hyperpolarized the resting membrane potential (RMP) by 2.9 +/- 0.4 mV (n = 9, P < 0.05). In the presence of HCO3-, the anion blocker SITS depolarized RMP by 2.6 +/- 0.5 mV (n = 5, P < 0.05). No HCO3--induced hyperpolarization was observed in the absence of extracellular Na+. The duration of the action potential measured at 50 % of repolarization time (APD50) was 29.2 +/- 6.1 % shorter in the presence of HCO3- than in its absence (n = 6, P < 0.05). 3. Quasi-steady-state currents were evoked by voltage-clamped ramps ranging from -130 to +30 mV, during 8 s. The development of a novel component of Na+-dependent and Cl--independent steady-state outward current was observed in the presence of HCO3-. The reversal potential (Erev) of the Na+-HCO3- cotransport current (INa,Bic) was measured at four different levels of extracellular Na+. A HCO3-:Na+ ratio compatible with a stoichiometry of 2:1 was detected. INa,Bic was also studied in isolation in standard whole-cell experiments. Under these conditions, INa,Bic reversed at -96.4 +/- 1.9 mV (n = 5), being consistent with the influx of 2 HCO3- ions per Na+ ion through the Na+ HCO3- cotransporter. 4. In the presence of external HCO3-, after 10 min of depolarizing the membrane potential (Em) with 45 mM extracellular K+, a significant intracellular alkalinization was detected (0.09 +/- 0. 03 pH units; n = 5, P < 0.05). No changes in pHi were observed when the myocytes were pre treated with the anion blocker DIDS (0.001 +/- 0.024 pH units; n = 5, n.s.), or when exposed to Na+-free solutions (0.003 +/- 0.037 pH units; n = 6, n.s.). 5. The above results allow us to conclude that the cardiac Na+-HCO3- cotransport is electrogenic and has an influence on RMP and APD of rat ventricular cells. PMID- 9729625 TI - Vagally evoked synaptic currents in the immature rat nucleus tractus solitarii in an intact in vitro preparation. AB - 1. Whole-cell voltage-clamp recordings in an in vitro brainstem-cranial nerve explant preparation were used to assess the local circuitry activated by vagal input to nucleus tractus solitarii (NTS) neurones in immature rats. 2. All neurones that responded to vagal stimulation displayed EPSCs of relatively constant latency. Approximately 50 % of these also demonstrated variable-latency IPSCs, and approximately 31 % also displayed variable-latency EPSCs to vagal stimulation. All neurones also had spontaneous EPSCs and IPSCs. 3. Evoked and spontaneous EPSCs reversed near 0 mV and were blocked by the glutamate AMPA/kainate receptor antagonists 6,7-nitroquinoxaline-2,3-dione (DNQX) or 6 cyano-7-nitroquinoxaline-2,3-dione (CNQX) at rest. Evoked EPSCs had rapid rise times (< 1 s) and decayed monoexponentially (tau = 2. 04 +/- 0.03 ms) at potentials near rest. 4. At holding potentials positive to approximately -50 mV, a slow EPSC could be evoked in the presence of DNQX or CNQX. This current peaked at holding potentials near -25 mV and was blocked by the NMDA receptor antagonist DL-2-amino-5-phosphonovaleric acid (AP5). It was therefore probably due to activation of NMDA receptors by vagal afferent fibres. 5. Fast IPSCs reversed near -70 mV and were blocked by the GABAA receptor antagonist bicuculline. In addition, bicuculline enhanced excitatory responses to vagal stimulation and increased spontaneous EPSC frequency. Antagonists to AMPA/kainate receptors reversibly blocked stimulus-associated IPSCs and also decreased the frequency of spontaneous IPSCs. 6. These findings suggest that glutamate mediates synaptic transmission from the vagus nerve to neurones in the immature NTS by acting at non-NMDA and NMDA receptors. NTS neurones may also receive glutamatergic and GABAergic synaptic input from local neurones that can be activated by vagal input and/or regulated by amino acid inputs from other brainstem neurones.1. Whole-cell voltage-clamp recordings in an in vitro brainstem-cranial nerve explant preparation were used to assess the local circuitry activated by vagal input to nucleus tractus solitarii (NTS) neurones in immature rats. PMID- 9729626 TI - Multimodal output mapping of human central motor representation on different spatial scales. AB - 1. Non-invasive mapping by focal transcranial magnetic stimulation (TMS) is frequently used to investigate cortical motor function in the intact and injured human brain. We examined how TMS-derived maps relate to the underlying cortical anatomy and to cortical maps generated by functional imaging studies. 2. The centres of gravity (COGs) of TMS maps of the first dorsal intersosseus muscle (FDI) were integrated into 3-D magnetic resonance imaging (MRI) data sets in eleven subjects. In seven of these subjects the TMS-derived COGs were compared with the COG of regional cerebral blood flow increases using positron emission tomography (PET) in an index finger flexion protocol. 3. Mean TMS-derived COG projections were located on the posterior lip of the precentral gyrus and TMS derived COG projections were in close proximity to the mean PET-derived COG, suggesting that the two methods reflect activity of similar cortical elements. 4. Criteria for a reliable assessment of the COG and the number of positions with a minimum amplitude of two-thirds of the maximum motor-evoked potential (T3Ps) were determined as a function of the number of stimuli and extension of the stimulation field. COGs and T3Ps were compared with an estimate of the size of the human motor cortex targeting alpha-motoneurons of forearm muscles. This comparison suggests that TMS can retrieve spatial information on cortical organization below the macroanatomic scale of cortical regions. 5. Finally, we studied the cortical representation of hand muscles in relation to facial and foot muscle representations and investigated hemispherical asymmetries. We did not find any evidence for a different ipsi- or contralateral representation of the mentalis muscle. Also, no difference was found between FDI representations on the dominant versus the non-dominant hemisphere. PMID- 9729627 TI - Single channel currents at six microsecond resolution elicited by acetylcholine in mouse myoballs. AB - 1. A patch-clamp set-up was optimized for low noise and high time resolution. An Axoclamp 200B amplifier was modified to incorporate a Teflon connector to the electrode. An electrode puller was equipped with a hydrogen-oxygen burner to produce quartz-glass pipettes with optimally 0.2 micron openings and 20 MOmega resistance. 2. The r.m.s. (root mean square) noise of sealed pipettes in the bath ranged from 3.6 fA with 100 Hz filter cut-off to 1.5 pA with 61 kHz filter cut off. At these extremes currents of 17 fA and more than 3 ms, or 9 pA and more than 6 micros could be resolved with a negligible error rate. 3. The system was tested on mouse myoballs, recording 9-10 pA single channel currents on-cell at 200 mV polarization which were elicited by 0.1-5000 microM acetylcholine (ACh). 4. Distributions of open and closed times and of correlations of open times to the preceding closed time defined several open states: single openings with mean durations of 1.2 and 25 micros, from single-liganded receptors, and bursts of 10 ms mean duration containing on average 800 micros openings and 16 micros closings, from double liganded receptors. Above 0.1 mM ACh these openings are interrupted increasingly by on average 18 micros and 72 micros channel blocks by ACh. PMID- 9729628 TI - Frequency-dependent synaptic depression modifies postsynaptic firing probability in cats. AB - 1. The influence of stimulus trains applied to single I a axons on the firing behaviour of single motoneurones was assessed in anaesthetized cats. The change in motoneurone firing probability associated with a single I a afferent spike was measured from short-latency peaks in peristimulus time histograms or cross correlograms. Some synapses showed frequency-dependent depression of the short latency peak, which is consonant with the frequency-dependent depression reported for the I a-motoneurone excitatory postsynaptic potential (EPSP). 2. Where they could be measured, EPSPs superimposed on the depolarizing ramps of potential recorded from motoneurones as they fired repetitively showed frequency-dependent changes in amplitude that parallelled those of the simultaneously recorded histograms. 3. Thus it appears that at synapses with small EPSPs, which are typical in the mammalian CNS, modulation of the EPSP should result in similar modulation of cell firing. PMID- 9729629 TI - Endogenous peptidergic modulation of perisynaptic Schwann cells at the frog neuromuscular junction. AB - 1. Although peptides are important modulators of synapses, their action on synapse-glia interactions remain unclear. The amphibian neuromuscular junction (NMJ) was used to examine the effects of substance P (SP) on perisynaptic Schwann cells (PSCs), glial cells at the frog NMJ, by monitoring changes in intracellular Ca2+. 2. SP induced Ca2+ responses that were mimicked by the neurokinin 1 receptor (NK-1) agonist septide and with a shorter delay by the SP fragment, SP(6 11). SP and SP(6-11) responses were blocked by NK-1 antagonists SR140333 and LY303870. 3. Ca2+ responses remained unchanged when extracellular Ca2+ was removed but were blocked after pertussis toxin (PTX) treatment, indicating that the receptors were linked to internal stores of Ca2+ via a PTX-sensitive G protein. 4. The slowly hydrolysable NK-1 agonist [Sar9, Met(O2)11]-SP only induced Ca2+ responses when applied for a long period of time and not during brief, local applications, suggesting the involvement of SP hydrolysis. Acetylcholinesterase (AChE) may not be involved in SP degradation since Ca2+ responses evoked by SP were unchanged in the presence of the cholinesterase inhibitor neostigmine. 5. Ca2+ responses induced by muscarine and nerve stimulations were almost abolished when preceded by SP applications, while those induced by ATP were significantly reduced. The rundown of the nerve-evoked Ca2+ responses in PSCs was attenuated in the presence of SR140333. 6. These results indicate that endogenous SP is involved in the regulation of PSC activity and that SP is an important modulator of glial cell Ca2+ signalling and synapse-glia communication. PMID- 9729630 TI - Tetrodotoxin-resistant impulses in single nociceptor nerve terminals in guinea pig cornea. AB - 1. Extracellular recording techniques have been used to study nerve impulses in single sensory nerve terminals in guinea-pig cornea isolated in vitro. 2. Nerve impulses occurred spontaneously and were evoked by electrical stimulation of the ciliary nerves. 3. The nerve impulses were identified as originating in polymodal receptors, mechano-receptors or 'cold' receptors. All three types are believed to be nociceptors. 4. Tetrodotoxin (TTX, 1 microM) blocked nerve impulses evoked by electrical stimulation of the ciliary nerves. However, ongoing and/or naturally evoked nerve impulses persisted in the presence of TTX in all three types of receptors. Lignocaine (lidocaine; 1 mM) blocked all electrical activity. 5. TTX resistant sodium channels therefore play a major role in generating the action potentials that signal pain to the brain. PMID- 9729631 TI - Potentiation of transmitter release by protein kinase C in goldfish retinal bipolar cells. AB - 1. We examined whether transmitter release could be modified by the activation of protein kinase C (PKC) of retinal bipolar cells. A bipolar cell with a large axon terminal was isolated from the goldfish retina. The presynaptic Ca2+ current was measured under whole-cell voltage clamp, and the released transmitter (probably glutamate) was detected electrophysiologically by using the response of NMDA receptors of catfish horizontal cells as a reporter. 2. Transmitter release was potentiated by a PKC activator, phorbol 12-myristate 13-acetate (PMA), but not by an ineffective phorbol ester, 4alpha-phorbol 12,13-didecanoate. A PKC inhibitor, bisindolylmaleimide I, did not affect the transmitter release by itself but blocked the PMA-induced potentiation of transmitter release. These results suggest that the actions of PMA were mediated via the activation of PKC. 3. Introduction of 5 mM EGTA into the presynaptic terminals of bipolar cells revealed two separate components of transmitter release. A rapid component was triggered immediately after depolarization while a slow component appeared with a delay. Application of PMA selectively potentiated the slow component without affecting the Ca2+ dependence of exocytosis. 4. We suggest that the activation of PKC may modify the recruitment process of synaptic vesicles in retinal bipolar cells. PMID- 9729632 TI - Interaction of platelet-activating factor, spleen and atrial natriuretic peptide in plasma volume regulation during endotoxaemia in rats. AB - 1. We studied endotoxin (lipopolysaccharide, LPS)-induced platelet-activating factor (PAF) production in various visceral organs, and the effect of PAF antagonists or splenectomy on LPS-induced changes. 2. PAF production in response to LPS was highest in the spleen, followed by ileum, heart, lung and kidneys. None was found in the liver. The splenic response was rapid, reaching 10 times the basal level at 30 min. The increased PAF content in each organ was unrelated to the enzyme activity of either macrophages or neutrophils. 3. LPS-induced hypotension and haemoconcentration were largely prevented by PAF antagonists and splenectomy. 4. Plasma volume fell, and plasma atrial natriuretic peptide (ANP) rose, after LPS administration. Splenectomy or pretreatment with PAF antagonists almost completely prevented these LPS-induced changes at 30 min, but only partially reversed them at 90 min. 5. These results suggest that during endotoxaemia: (a) the spleen is the site of the highest endogenous PAF production; (b) the initial release of ANP is dependent on the production of endogenous PAF, and a PAF-ANP interaction mediates the early plasma volume reduction; (c) plasma volume reduction as well as ANP release depend on the spleen; (d) PAF mediated the hypotensive response and its action in the spleen; and (e) sequestered neutrophils are probably not the main source of PAF in the spleen. PMID- 9729633 TI - Monocularly programmed human saccades during vergence changes? AB - 1. When binocular fixation is shifted to a new target located at a different distance and in a different direction from initial fixation, a binocularly unbalanced saccade occurs at or near the onset of the composite eye movement. Those saccades typically produce good post-saccadic foveation of the target by one eye or the other. 2. Following such saccades, the better-aligned eye is typically as well aimed at the target as after pure versional saccades, but the partner eye deviates much more, thus requiring asymmetrical post-saccadic vergence movement. 3. This phenomenon suggests that during binocular viewing, the retinal eccentricity of a new-target's image from one of the eyes can be used to programme that eye's own saccade, so that it arrives reliably on target; and that the images of that target from both eyes participate in generating the saccadic excursion of the partner eye. 4. The ecologically useful result is rapid achievement of a high-resolution monocular view of the new target, although full binocular foveation is achieved much later. PMID- 9729634 TI - Muscle shortening induced by tenotomy does not reduce activity levels in rat soleus. AB - 1. A slow postural muscle was tenotomized to determine the role of muscle stretch on chronic recruitment patterns in freely moving animals. 2. Different amounts of muscle shortening were induced in the soleus muscles of ten rats by severing the tendon of insertion (n = 3), the whole Achilles' tendon (n = 4) or the origins and insertions (n = 3). 3. Bipolar wire electrodes were implanted on each muscle to record the electromyographic activity (EMG) under control and tenotomized conditions. The complex interference pattern was continuously analysed to determine the number and amplitude of peak potentials (called turns). The numbers of these 'turns' and their amplitudes were determined during 4 control and at least 5 experimental days. Sham-operated controls and groups matched according to the type of tenotomy were analysed for length changes and pathological changes 5 and 10 days post-tenotomy. 4. The total activity levels in all three tenotomy conditions were not significantly changed when compared with their own control levels. No differences in total activity level were found between the three tenotomized conditions. 5. The normal diurnal patterns of muscle recruitment were preserved during the tenotomized conditions, with the highest levels consistently occurring during the first 3 h of the dark cycle. 6. Tenotomy of the soleus, whether induced by distal (ST), distal and proximal (DT) or Achilles' tenotomy (AT) resulted in muscle shortening (9-26 %). No muscle pathology was found in the ST or AT groups. Degeneration was found in the DT group after 5 days, with further increases at 10 days. 7. These data suggest that the absence of stretch had no discernible influence on the aggregate activity levels in the slow postural soleus muscle. Whether tenotomy caused changes in recruitment within individual step cycles was not evaluated. PMID- 9729635 TI - Evidence that a transcortical pathway contributes to stretch reflexes in the tibialis anterior muscle in man. AB - 1. In human subjects, stretch applied to ankle dorsiflexors elicited three bursts of reflex activity in the tibialis anterior (TA) muscle (labelled M1, M2 and M3) at mean onset latencies of 44, 69 and 95 ms, respectively. The possibility that the later of these reflex bursts is mediated by a transcortical pathway was investigated. 2. The stretch evoked a cerebral potential recorded from the somatosensory cortex at a mean onset latency of 47 ms in nine subjects. In the same subjects a compound motor-evoked potential (MEP) in the TA muscle, evoked by magnetic stimulation of the motor cortex, had a mean onset latency of 32 ms. The M1 and the M2 reflexes thus had too short a latency to be caused by a transcortical pathway (minimum latency, 79 ms (47 + 32)), whereas the later part of the M2 and all of the M3 reflex had a sufficiently long latency. 3. When the transcranial magnetic stimulation was timed so that the MEP arrived in the TA muscle at the same time as the M1 or M2 reflexes, no extra increase in the potential was observed. However, when the MEP arrived at the same time as the M3 reflex a significant (P < 0.01) extra-facilitation was observed in all twelve subjects investigated. 4. Peaks evoked by transcranial magnetic stimulation in the post-stimulus time histogram of the discharge probability of single TA motor units (n = 28) were strongly facilitated when they occurred at the same time as the M3 response. This was not the case for the first peaks evoked by electrical transcranial stimulation in any of nine units investigated. 5. We suggest that these findings are explained by an increased cortical excitability following TA stretch and that this supports the hypothesis that the M3 response in the TA muscle is - at least partly - mediated by a transcortical reflex. PMID- 9729636 TI - The effect of repeated acute hypoxaemia on fetal cardiovascular development in the sheep. AB - 1. Hypoxaemia during intrauterine life may be important in the development of cardiovascular diseases in later life. Thus it was the aim of this study to investigate the effect of repeated acute hypoxia on the cardiovascular development and growth of the fetus. 2. Fourteen fetal sheep (105-109 days gestational age) were instrumented with amniotic and vascular catheters, an electrocardiogram (ECG) electrode and a flow probe around the femoral artery. Seven animals were given repeated acute isocapnic hypoxaemia (Pa,O2 reduced to ca. 13 mmHg) for 1 h every day for 14 days and they were compared to seven animals which remained normoxic throughout with respect to fetal mean arterial blood pressure (MAP), fetal heart rate (FHR), and fetal baro- and chemoreflexes. 3. No differences were found between the two groups of fetuses in FHR, MAP, baro- or chemoreflexes, femoral blood flow, femoral vascular resistance or fetal growth. 4. Repeated acute hypoxaemia of a moderate degree over a period of 2 weeks in late gestation does not affect cardiovascular development or growth in the fetal sheep. PMID- 9729637 TI - Epiglottic movements during breathing in humans. AB - 1. Using X-ray fluoroscopy we measured antero-posterior (A-P) and cranio-caudal (C-C) displacements of the epiglottic tip (ET), corniculate cartilage and hyoid bone in seven seated, normal human subjects (age 34 +/- 3 years; mean +/- S.E.M.; 4 males, 3 females) breathing via a nasal mask or mouthpiece with (RL) and without (UB) a fixed resistive load. 2. During UB, via either mouth or nose, there were no significant A-P ET movements. During RL via the nose the ET at peak expiratory flow was 2.6 +/- 1.3 mm cranial to its position at peak inspiratory flow (P < 0.05, ANOVA). C-C movements of the ET correlated strongly with C-C movements of the corniculate cartilage and hyoid bone. 3. The ET, corniculate cartilage and hyoid bone (at zero airflow) were situated more caudally during oral UB than for any other condition. 4. When present, epiglottic movements during breathing do not appear to be independent of those of the larynx and hyoid. Furthermore, epiglottic position may be related to the level of upper airway resistance. PMID- 9729638 TI - Cutaneous receptive fields and topography of mossy fibres and climbing fibres projecting to cat cerebellar C3 zone. AB - 1. The topographical organization of mossy fibre input to the forelimb area of the paravermal C3 zone in cerebellar lobules IV and V was investigated in barbiturate-anaesthetized cats and compared with the previously described microzonal organization of climbing fibre input to the same part of the cortex. Recordings were made in the Purkinje cell and granule cell layers from single climbing fibre and mossy fibre units, respectively, and the organization of cutaneous receptive fields was assessed for both types of afferents. 2. Based on spatial characteristics, receptive fields of single mossy fibres could be systematized into ten classes and a total of thirty-two subclasses, mainly in accordance with a scheme previously used for classification of climbing fibres. Different mossy fibres displayed a substantial range of sensitivity to natural peripheral stimulation, responded preferentially to phasic or tonic stimuli and were activated by brushing of hairs or light tapping of the skin. 3. Overall, mossy fibres to any given microzone had receptive fields resembling the climbing fibre receptive field defining that microzone. However, compared with the climbing fibre input, the mossy fibre input had a more intricate topographical organization. Mossy fibres with very similar receptive fields projected to circumscribed cortical regions, with a specific termination not only in the mediolateral, but also in some cases in the rostrocaudal and dorsoventral, dimensions of the zone. On the other hand, mossy fibre units with non-identical, albeit usually similar, receptive fields were frequently found in the same microelectrode track. PMID- 9729639 TI - Activity of alkaline phosphatase in the major salivary glands of mice at various ages of postnatal life, and during pregnancy and lactation. AB - Activity of alkaline phosphatase in the major salivary glands of male and female mice at various ages of postnatal life, and in females during lactation was studied histochemically. Enzyme activity was not detected on the day of birth, but was found in the terminal tubules of all major salivary glands during the first postnatal week. Alkaline phosphatase activity was increasing gradually with age and a definitive enzymatic pattern was observed by the age of 6 weeks. No difference in enzyme activity was found among the major salivary glands of young adult and old animals. The parenchyma of fully differentiated submandibular glands showed clear sexually dimorphic patterns of alkaline phosphatase activity. During pregnancy, a significant increase of alkaline phosphatase activity was detected in submandibular gland. From gestation day 15 to the end of pregnancy, enzymic pattern of granular convoluted tubules of pregnant females was the same as in the adult males. Histochemical masculinization of the submandibular gland during pregnancy suggests that besides androgens also progesterone exerts masculinization of the murine submandibular salivary gland. PMID- 9729640 TI - Immunohistochemical detection of intermediate filament nestin. AB - Using Rat-401 monoclonal antibody and peroxidase immunohistochemistry we have detected IF nestin in developing and adult rat tissues. Although epitope recognized by Rat-401 antibody is relatively resistant to aldehyde fixation and paraffin embedding, the embedding of tissue samples into polyester wax and microwave antigen retrieval of histological sections enabled us to enhance sensitivity of immunohistochemical detection and to identify cells expressing low levels of nestin. Our findings confirm that nestin is predominantly distributed in developing neural, myogenic and mesenchymal cells, i.e. cell types that have been previously described to express this intermediate filament. Furthermore, we made original findings on identification of nestin expression in additional cell types, e.g. newly formed endothelial cells of extra- and intraembryonic blood vessels, epithelial cells of the developing lens, and cells apposed to to hair follicles. PMID- 9729641 TI - Ambivalent effect of long lasting terguride treatment on genetically based glycide and lipid metabolism abnormalities in SHR/N-cp Koletsky rats. AB - Experiments were carried out in the genetically hypertensive obese rats of Koletsky type (SHR/N-cp obese), in their lean siblings (SHR/N-cp lean) and in the rats of Han-Wistar strain. The effect of long lasting terguride treatment was monitored when the animal represents a control for itself. Blood was sampled to heparinized capillaries from retrobulbar plexus in the same animal before and after the terguride treatment. Long lasting terguride treatment shows decrease in "area under the glucose tolerance curve" in all groups of animals, increase in glycaemia in normotensive males, increase in insulinemia in normotensive rats of both sexes and in SHR/N-cp lean males and decrease of insulinemia in SHR/N-cp obese males, increase of triglyceridemia in normotensive rats of both sexes and in SHR/N-cp lean males, and decrease of triglyceridemia in SHR/N-cp lean females and SHR/N-cp obese of both sexes, increase in cholesterolemia in normotensive and SHR/N-cp lean males, decrease in cholesterolemia in normotensive, SHR/N-cp lean and obese females. Thus ambivalent effect of terguride treatment is more expressive in glucose tolerance and in plasma triglycerides. Ambivalent effect of long lasting terguride treatment is profoundly expressed in correlation between pre-treatment state of individual parameters and the effect of treatment expressed in percentage changes in post-treatment state. In all cases statistically significant correlation coefficients show negative mark. Thus it is apparent that terguride increases monitored parameter when this parameter is low before treatment, and vice versa. When it is considered "area under the glucose tolerance curve", significance was attained in all groups, when judging basal glycaemia, significance was attained in normotensive and SHR/N-cp lean rats of both sexes, taking into account insulinemia, significance was attained in normotensive and SHR/N-cp obese rats of both sexes, when analysing plasma triglycerides, significance was attained in normotensive rats of both sexes and in SHR/n-cp lean females and obese males, when we consider total plasma cholesterol, significance was attained in normotensive rats of both sexes and in SHR/N-cp lean males. From the clinical point of view it must be underlined that terguride is potent to increase insulinemia. Thus there is open a possibility of the other clinical indication of the mentioned drug. PMID- 9729642 TI - Hypocholesterolemic effect of pravastatin is associated with increased content of antioxidant vitamin-E in cholesterol fractions. AB - Metabolic studies support the findings that antioxidants inhibit atherosclerosis. Treatment with vitamin E reduced both the susceptibility of low density lipoprotein cholesterol (LDL-C) to in vivo lipid peroxidation and atherosclerosis and smooth muscle proliferation. Thus the aim of present study was to examine metabolic consequences of reduced plasma LDL-C during hypolipidemic therapy and the distribution of antioxidant vitamin E. A group of 10 patients (4 men, 6 women, age 35-65y) with familial hypercholesterolaemia was treated using pravastatin (Lipostat Bristol Myers Squibb, 40 mg daily at 6:00 PM). Blood samples were examined before treatment, after 4 and 8 weeks of therapy. After ultracentrifugation, samples were analyzed for lipoprotein fractions and the content of vitamin E and cholesterol. Pravastatin reduced both total cholesterol (9.85 +/- 0.74 vs. 6.81 +/- 0.51 mmol/1; p < 0.01), LDL-C (6.42 +/- 0.45 vs. 4.51 +/- 0.45 mmol/l; p < 0.01), light LDL1-C (4.56 +/- 0.50 vs. 3.11 +/- 0.34 mmol/l; p < 0.05) and dense LDL2-C (1.86 +/- 0.27 vs. 1.42 +/- 0.17 mmol/l; ns). Serum vitamin E was reduced during hypolipidemic therapy in the fraction of total, LDL1, LDL2, and VLDL-cholesterol. However, the ratio of serum vitamin E/total serum cholesterol (4.57 +/- 0.32 vs. 5.12 +/- 0.37 mmol/l/mmol/l; p < 0.05) and ratio of LDL2-C vitamin E/LDL2-C (3.92 +/- 0.07 vs. 4.64 +/- 0.37 mmol/l/mmol/l; p = 0.08) increased in comparison to pre-treatment values. We conclude that pravastatin therapy may possess anti-atherogenic properties which involve not only its hypocholesterolemic effect, but also its favorable effects on the distribution of LDL subclasses and the content of antioxidant vitamin E in atherogenic lipoproteins. PMID- 9729643 TI - How long can the previously assembled cardiopulmonary bypass circuit stay sterile? AB - The sterility of previously assembled cardiopulmonary bypass circuits was investigated for 100 extracorporeal circuits. The closed circuits were assembled using aseptic technique and remained in the pump room until time of use. The mean time from point of setup to point of priming for the 100 consecutive circuits was 32 hours, with a range of 19 to 89 hours. Circuits were primed with the calculated volume of priming solution, circulated for 5 minutes and tested for microbial contamination by withdrawing 20 ml of the priming solution and 10 days incubated in Thioglycolate and Sabouraud culture mediums. All were found to be free of microbial contamination. The results of this investigation demonstrate that the sterility of the extracorporeal circuit, pre-assembled in advance of actual priming, can be maintained over an extended interval when standard aseptic technique is used. This allows the utilization of a pre-assembled circuit for emergency cardiopulmonary support. PMID- 9729644 TI - Femoral interlocking nailing: a review of the experience at Chang Gung Memorial Hospital. AB - The invention of the interlocking nail has broadly extended the indications for reamed intramedullary nailing. It has even become the treatment of choice for most complex femoral shaft fractures. At Chang Gang Memorial Hospital, about 180 femoral cases have been treated with this technique each year since December 1986. With this accumulation of experience, both theoretical and clinical advancements are continuously developing and are regularly reported in the literature. In this article, experiences in the past few years with this technique are reviewed and evaluated. The author believes that to be a good trauma orthopedist, through training in femoral interlocking nailing, both theoretically and technically, is absolutely imperative. This device does, however, still have some disadvantages, but further improvements are anticipated with the advance of modern science. PMID- 9729645 TI - Serum-soluble interleukin-2 receptor concentrations in patients with breast cancer: a preliminary report. AB - BACKGROUND: The serum-soluble interleukin-2 receptor has been claimed to be a marker of host biological response in patients with solid malignancies. This study was designed to evaluate the biological significance of the preoperative serum-soluble interleukin-2 receptor concentration in patients with invasive breast cancer. MATERIALS AND METHODS: Venous blood samples were collected from 66 patients with invasive breast carcinoma and the serum concentrations of soluble interleukin-2 receptor were measured with an enzyme immunoassay method. Data regarding maximum tumor diameter, age, estrogen receptor status, lymph node status, distant metastasis status, histologic grade, ploidy pattern and S-phase fraction were also collected and evaluated simultaneously with the serum concentration levels of soluble interleukin-2 receptor. Fifteen age-matched healthy subjects were used as a control group. RESULTS: The mean value of serum soluble interleukin-2 receptor in patients with invasive breast cancer was 621 +/ 145 units/ml and that of the control group was 308 +/- 59 units/ml; and the difference was significant (p = 0.000). With multivariate analysis, lymph node status (p = 0.000), distant metastasis status (p = 0.000) and maximum tumor diameter (p = 0.000) appeared as independent factors in regards to the significantly, higher serum concentrations of soluble interleukin-2 receptor. CONCLUSION: Based on our preliminary results, the preoperative serum-soluble interleukin-2 receptor concentration is closely related to lymph node status, distant metastasis status and tumor diameter in invasive breast carcinoma. This may be an additional valuable predictive factor for the diagnosis of invasive breast cancer. PMID- 9729646 TI - Development of a dual-probe detection system for positron emission radiotracer detection. AB - BACKGROUND: Positron emission radiotracers are very useful in biochemical and pharmacological research. The purpose of this work was to develop and build a simple and inexpensive dual-probe detector system (DPDS) for detecting the activity of positron emission radiotracers. MATERIALS AND METHODS: The design of the DPDS was based on the coincident detection of two 511 KeV annihilation gamma rays, which determined the positron activity in the body. A pair of sodium iodide (NaI) scintillators coupled to a photomultiplier tube (PMT) were positioned face to-face as in a dual-probe system. The signals from each probe were then sent to a preamplifier, a linear amplifier, a single channel analyzer (SCA), and the primary and secondary coincidence units, respectively, to determine the number of total coincidence events and random events. Validation of basic characteristics, including stability, sensitivity, linearity and geometric counting efficiency, were tested. RESULTS: The sensitivity and linearity of the DPDS worked well in detecting radioactivity within 100 microCi. The DPDS was very stable in continuous counting for more than 2 hours. The geometric counting efficiency changed less than 5% when the source was placed 2 cm from the central coincidence line. CONCLUSION: This DPDS is a promising tool for positron emission radiotracer detection. It may be helpful in future in both clinical and basic biomedical research, such as neuroreceptor occupancy or glucose metabolic studies. Eventually, we will be able to use the DPDS in the human or animal body for evaluating position emission radiotracer metabolism and distribution. PMID- 9729647 TI - Continuous venovenous hemodialysis in multiple organ systems failure patients with acute renal failure. AB - BACKGROUND: Continuous renal replacement therapies have become the preferable methods for treating critically ill patients with acute renal failure. We studied the use of continuous venovenous hemodialysis (CVVHD) for the management of multiple organ failure patients with acute renal failure. MATERIALS AND METHODS: Forty patients in intensive care units were studied. Their mean age was 50.5 (range 21 to 82) years. The mean of number of organ system failures per patient was 4.9 (range 4 to 6). All patients required mechanical ventilation. CVVHD was performed from the femoral vein via a double-lumen catheter. Blood flow was maintained via blood pump at rate of 120 to 150 ml/min. Bicarbonate solution was delivered to the dialysate ports of the filter at rate of 700 to 900 ml/hour. Any complications during therapy were recorded. RESULTS: The mean urea clearance was 14.7 ml/min and the mean creatinine clearance was 15.4 ml/min. Electrolytes were all within the normal ranges. There were two episodes of extracorporeal circuit blood clotting and one patient experienced hypothermia. Seven of the 40 patients (17.5%) survived. CONCLUSION: CVVHD is a safe, effective, easily established therapy for managing the multiple organ failure patients with acute renal failure, offering good metabolic, electrolyte, and fluid control. PMID- 9729648 TI - Comparison of cerebral occipital gray and parietal white matter metabolite differences using PRESS and STEAM at different echo times. AB - BACKGROUND: Reports regarding the differences in metabolite ratios with different echo times (TEs) at various brain regions are rare. The purpose of this study is to investigate in Chinese the cerebral metabolite ratios in gray matter (GM) and white matter (WM) using both STEAM and PRESS techniques at TEs of 30 ms and 136 ms, respectively. MATERIALS AND METHODS: Using GE Signa 1.5 T Scanner with STEAM and PRESS sequences, automated single voxel localized proton magnetic resonance spectroscopy (1H-MRS) were applied to identify the metabolite ratio differences in cerebral parietal white and occipital gray matter of healthy volunteers. RESULTS: Metabolite ratios between GM and WM were significantly different in N acetyl aspartate/creatine (Cr), choline/Cr and myo-inositol/Cr. There were also significant differences in metabolite ratios when different echo times were applied. CONCLUSION: Reliable metabolite ratios at different brain regions can be obtained using automated data acquisition and spectral processing in single voxel localized 1H-MRS. This technique may remove user-dependent bias and provide more efficient and consistent ways to analyze the spectral data. However, one must be careful in interpreting spectroscopic data based on different regions and acquisition parameters. PMID- 9729649 TI - Spontaneous cellulitis in adults with idiopathic nephrotic syndrome. AB - BACKGROUND: Susceptibility to infection is a common problem in a patient with nephrotic syndrome. The spontaneous cellulitis is not uncommon in pediatric patients with nephrotic syndrome, whereas there have been few cases reported in adults. In order to clarify the clinical course of this complication, we present 17 adult idiopathic nephrotic patients with spontaneous cellulitis. MATERIALS AND METHODS: A series of 17 adult idiopathic nephrotic patients with spontaneous cellulitis were retrospectively reviewed in Chang Gung Memorial Hospital, Kaohsiung from 1986 through 1996. We analyzed their physical conditions, clinical manifestations, and outcome. All patients received renal biopsies and had pathologic diagnoses. RESULTS: The medical records of 17 patients were collected, 12 men and 5 women, with ages ranging from 16 to 63 years (mean 29.5 years). The pathologic diagnoses of renal biopsies included minimal change disease (13/17), membranous glomerulonephritis (2/17), mesangioproliferative glomerulonephritis (1/17) and focal/segmental glomerulosclerosis (1/17). All patients had generalized edematous state. The clinical presentations of these patients were variable. The mean serum albumin and daily urinary protein excretion were 1.28+/ 0.64 g/dl and 8.75+/-5.16 g, respectively. The results of blood cultures were E. coli (3/17), Gram-negative bacilli (1/17), Streptococcus viridans (1/17), Streptococcus pneumoniae (1/17) and no growth (11/17). All patients responded to antibiotic treatment except one patient who died due to overwhelming sepsis. CONCLUSION: The related factors of spontaneous cellulitis in patients with nephrotic syndrome are edematous skin, hypoalbuminemia, immunosuppressive drugs and defective immunity. Our patients had accordant conditions. The prognosis was good if diagnosis and treatment are made early. PMID- 9729650 TI - Chronic refractory tibia osteomyelitis treated with adjuvent hyperbaric oxygen: a preliminary report. AB - BACKGROUND: Refractory osteomyelitis is a serious disease that fails to respond to aggressive medical and surgical treatment. A plethora of alternative therapies have evolved. Hyperbaric oxygen has been proven to enhance bone and soft tissue healing in many in vitro and in vivo studies. This article presents the preliminary results of adjunctive hyperbaric oxygen therapy in patients with refractory osteomyelitis. MATERIALS AND METHODS: Fifteen patients who were diagnosed with refractory tibia osteomyelitis were treated prospectively with adjunctive hyperbaric oxygen therapy, aggressive surgical debridement, and parenteral antibiotic treatment. The effectiveness was evaluated with an average follow-up of 17.2 months. RESULTS: The hyperbaric oxygen therapy averaged 26 daily sessions. Successful treatment was achieved in 13 patients (86%). The mean length of treatment was 45 days. The preliminary results are comparable with other series. CONCLUSION: Hyperbaric oxygen is effective as an adjunct to aggressive medical and surgical management in refractory osteomyelitis. A precise clinical staging system for patient selection and treatment organization is imperative to successful outcome. PMID- 9729651 TI - Drug use patterns and gender differences among heroin addicts hospitalized for detoxification. AB - BACKGROUND: There has been a surge of heroin abuse in Taiwan in recent years making it necessary to study and understand the characteristics, drug use patterns and behavior among heroin users. MATERIALS AND METHODS: Two hundred and eighty-three patients hospitalized for heroin detoxification received a diagnostic interview and a semi-structured interview which rendered the demographic information, medical history, and patterns of and reasons for heroin use. Differences between male and female drug users were also compared. RESULTS: More than half of the subjects (54.3%) were unemployed. The percentage of unemployment of female patients was significantly greater than that of male patients (75.9% vs. 48.0%, p<0.05). Women were significantly younger (p<0.001) and had a significantly earlier (p< 0.001) onset of heroin use than men. About one-third of the subjects (33.9%) were multiple drug users, with amphetamines as the most common (79.2%) concomitant drug of abuse. More men reported curiosity as the reason for first use, while more women reported peer influence as the reason for first use. CONCLUSION: This study showed that significant gender differences in employment status, age of first use, and reasons for drug use among heroin addicts. Further exploration of gender and cross-cultural differences could have important theoretical and treatment implications. PMID- 9729652 TI - Helicobacter pylori in surgical specimens from patients with resectable gastric adenocarcinoma. AB - BACKGROUND: To investigate the prevalence of Helicobacter pylori (H. pylori) in surgical specimens from patients with respectable gastric adenocarcinoma using a histological method, and to study the specific biology of H. pylori-associated gastric cancer. MATERIALS AND METHODS: The presence of H. pylori in resected specimens from 276 patients treated between October 1994 and October 1996 was evaluated histologically using hematoxylin-eosin stain. Clinicopathologic data, including age, gender, cancer location, invasion depth, histologic type (intestinal or diffuse), stage (early or advanced), and the histology of the noncancerous gastric mucosa, were compared between patients testing positive and those testing negative for H. pylori. RESULTS: The overall positive rate of H. pylori was 27.5%. H. pylori-positive gastric cancer was frequently associated with atrophic gastritis or intestinal metaplasia in the noncancerous tissue, but was not associated with other variables. CONCLUSION: The results contradict the prevailing concept that H. pylori rarely colonizes in the metaplastic mucosa. To determine whether patients who have normal mucosa with superficial gastritis have a lower seroprevalence of H. pylori, further serologic study of the IgG antibody against H. pylori is mandatory. Our histologically positive rate is relatively lower than the seropositive one. More tissue samplings and special stains may provide more precise results. PMID- 9729653 TI - Evidence of Epstein-Barr virus in lymphoepithelioma-like carcinoma of the uterine cervix: a case report. AB - Lymphoepithelioma-like carcinoma of the uterine cervix is very rare. A 71-year old woman, who had cervical cancer stage IB, presented lymphoepithelioma-like carcinoma pathologically. The neoplasm revealed an undifferentiated, nonkeratinizing carcinoma, which contained prominent vesicular nucleoli histologically. Epstein-Barr virus (EBV) DNA and HPV-16 DNA were demonstrated in cancer cells when using polymerase chain reaction. The patient underwent a radical hysterectomy with bilateral pelvic lymph node dissection and bilateral salpingooophorectomy. Her postoperative course was uneventful. The literature and pathologic findings are reviewed and discussed. PMID- 9729654 TI - Primitive neuroectodermal tumor of the lingual nerve: a case report. AB - This is a report of a primitive neuroectodermal tumor (PNET) arising from the lingual nerve of a 5-year-old girl. Twelve months after surgical excision and postoperative chemotherapy, the patient remained alive without any evidence of recurrence or metastasis. The occurrence of PNETs in the head and neck region in children is rare. This is the first case report in the literature of a PNET originating from a cranial nerve in a child. Because of its aggressive nature and poor prognosis, aggressive surgical and medical treatment regimens are needed. Surgeons should be mindful to include PNET in the differential diagnosis of head and neck tumors. PMID- 9729655 TI - Adrenal hemangioma: two cases report. AB - Adrenal hemangiomas are rare, nonfunctioning benign tumors. They are well circumscribed and comprise of closely adjacent vascular channels of varying sizes that are lined with a single layer of endothelium. When they occur, they are frequently located in the skin and liver. There are no characteristic symptoms of adrenal hemangioma unless the tumor reaches a size large enough to exert pressure. To our knowledge, there have been 14 clinical cases of adrenal hemangiomas reported, all with similar pathologic features. We present 2 additional cases, which were identified incidentally after non-urologic complaints (epigastric fullness and low back pain in patient 1 and patient 2, respectively). In these 2 patients, the tumors were surgically removed and diagnosed postoperatively as adrenal hemangiomas. Preoperative radiologic findings on plain film and abdominal computerized tomography showed the characteristic round calcifications with translucent centers, typical of phleboliths, to be pathognomonic of adrenal gland hemangiomas. Unfortunately, this characteristic feature of adrenal hemangioma existed in only one case report. In conclusion, the preoperative diagnosis of adrenal hemangioma is difficult but should be kept in mind as being part of the differential diagnosis of adrenal tumors. PMID- 9729656 TI - Endobronchial metastasis from an occult papillary thyroid carcinoma: a case report. AB - It is well known that occult papillary thyroid cancers can result in regional lymph node metastasis. However, this small, clinically undetectable cancer rarely causes distant metastasis; moreover, an endobronchial presentation has not been described previously. We report on a 48-year-old man who experienced hemoptysis. A fiberoptic bronchoscopic biopsy established the diagnosis of endobronchial metastasis in the absence of clinically apparent thyroid cancer. After the patient was treated with a total thyroidectomy and radioactive iodine (131I), a whole body radioiodine scan revealed no evidence of distant functioning metastasis. A histological evaluation of the thyroid gland showed the presence of a 0.2 x 0.2 x 0.2 cm nodule in the right lobe. A left pneumonectomy was undertaken to treat the distant metastasis of the disease. Since most papillary thyroid carcinomas are curable, an aggressive surgical approach to the solitary metastasis is indicated. PMID- 9729657 TI - Penicillium marneffei fungemia in an AIDS patient: the first case report in Taiwan. AB - We report a 36-year-old man with acquired immunodeficiency syndrome (AIDS), presenting systemic Penicillium marneffei (PM) infection. Fungal culture from the blood isolated PM. PM-induced enteritis was also suspected in this patient although there was no direct evidence. He also had other manifestations of immunocompromised status, including military tuberculosis and oral candidiasis. He died of respiratory failure in spite of prompt treatment for infection. This is the first confirmed case of PM infection in Taiwan. PMID- 9729658 TI - Early recognition of unsuspected malignant hyperthermia and successful management of serve myoglobinuric renal failure in subsequent rhabdomyolysis: a case report. AB - Malignant hyperthermia (MH) is a rare inherited disorder of skeletal muscle, its inheritance being autosomal dominant and its mutant gene located on chromosome 19. MH is occasionally observed during general anesthesia when using some special triggering agents. In the susceptible patient, it presents with a fulminant skeletal muscle hypermetabolic crisis and proceeds to serve rhabdomyolysis. Once rhabdomyolysis is established, acute renal failure is not an inevitable consequence, yet it is the fatal complication if the condition is not appropriately managed. We describe a case of acute renal failure in the setting of rhabdomyolysis in unsuspected MH, resulting in full recovery after intermittent hemodialysis. In this case, we emphasize the importance of early recognition of MH and the favorable prognosis of subsequent myoglobinuric renal failure if treated appropriately. PMID- 9729659 TI - Ultrasound recognition and treatment of fetal supraventricular tachycardia with hydrops: a case report. AB - To manage fetal tachyarrhythmia induced hydrops, both a correct diagnosis and adequate intrauterine therapy are fundamentally important. We present a 32-week gestational-age hydropic fetus with supraventricular tachycardia who responded dramatically after transplacental administration of high dose digoxin (1 mg intravenously daily). The baby was born at 36 weeks' gestation followed by a successful postnatal conversion. Prenatal fetal echocardiography is emphasized in determining appropriate treatment and monitoring fetal well-being which in this case resulted in a good outcome. PMID- 9729660 TI - Hypertrophic osteoarthropathy in nasopharyngeal carcinoma patients: two cases report. AB - Hypertrophic osteoarthropathy (HOA) is a rheumatic disorder characterized by digital clubbing, bone pain, and arthralgia. HOA can be idiopathic or secondary to a variety of pulmonary, cardiogenic, or malignant disorders. We present 2 male patients, aged 46 and 42, with advanced nasopharyngeal carcinoma (NPC) who developed HOA 1-4 years after radiotherapy. Differential diagnosis between HOA and coexisting bone metastasis must be made with caution. We found bone scintigraphy to be the most sensitive tool to distinguish between these 2 disease. Intense symmetrical uptake of radioisotope along the cortex of long bones, so-called parallel tract sign, is typical. Plain radiographs demonstrating prominent periosteal reaction were also effective for this. The rheumatic manifestation of HOA was paraneoplastic and related to pulmonary metastasis. The clinical manifestation of the 2 patients suggested that pulmonary metastasis should be suspected in NPC patients when HOA appears. PMID- 9729661 TI - Solitary osteochondroma of the lumbar spine with cord compression: a case report. AB - Solitary osteochondromas are usually of little significance. However, if they are located near neurologic structures, they may cause irritation due to compression. Spinal cord compression is a rare but potentially catastrophic manifestation of solitary osteochondromas. As far as can be ascertained, a solitary osteochondroma in the lumbar region with spinal cord compression has not been reported in Taiwan. This article includes the treatment of an 11-year-old girl with osteochondromas which was complicated by the late development of kyphoscoliosis and a review of the literature. PMID- 9729662 TI - Hepatocellular carcinoma presenting as jaundice, hemobilia, and acute pancreatitis: a case report. AB - A 48-year-old asymptomatic male hepatitis B virus carrier presented with a 2-day history of fever, chills, right upper quadrant abdominal pain, and jaundice. Shock was detected on admission. Emergency abdominal computed tomography (CT) scanning without contrast enhancement showed the features of acute pancreatitis. Hemobilia, edematous pancreatitis, cholestasis and cholecystitis were found on exploratory laparotomy. Neither stone nor active bleeding were detected on intraoperative choledochoscopic examination. Postoperative T-tube cholangiography one month later revealed non-opacification of the left intrahepatic duct. The patient's abdominal pain and hemobilia recurred. Celiac angiography and CT scanning with contrast showed two hepatocellular carcinomas (HCC) in the left lobe of the liver. This is the first case report in the English literature of HCC presenting as jaundice, hemobilia, and acute pancreatitis. PMID- 9729663 TI - [Arkovy and Hungarian dentistry]. PMID- 9729664 TI - [Smoking and diseases of the oral cavity]. AB - The paper gives a review on the etiology of oral cancer, discusses some of the mechanisms by which tobacco may bring about changes in oral tissues, and describe ways in which the dentist can help with the problem of tobacco use. Risk of developing oral cancer increases with the amount and length of cigarette-smoking and alcohol consumption. Hungry is ranked with the highest cigarette-consumption as third among 111 countries world-wide. Investigations of the authors on the pathogenesis of leukoplakia by tobacco, demonstrated increased tissue levels of inflammatory mediators, and stimulated keratinocyte proliferation. Finally, the dentist has an important role to play in society as an advocate for reducing tobacco use. PMID- 9729665 TI - [Root canal preparation for obturation using nickel-titanium mechanical devices]. AB - Engine-driven ProFile instruments manufactured by Maillefer (Switzerland) and Tulsa Dental Products (USA) have been used for cleaning and shaping root canals. Both of them are nickel titanium, U-shaped instruments designed for a controlled slow speed (250 rpm) rotary handpiece. In contrast to the Maillefer instruments ProFile Series 29 files (Tulsa Dental) show a constant 29% increase between tip diameters. As these more tapered instruments (ProFile 0.04; 0.06) are rotated, they produce a "crown-down" preparation, resulting in a tunnelform taper from orifice to apex. According to our experiences ProFile instruments are useful alternative tools for root canal preparation quickly and efficiently. PMID- 9729666 TI - [The role of sensory nerves in the development of inflammation of oral tissues]. AB - Experimental stimulation and clinical procedures applied on the tooth crown cause vascular reactions in the dental pulp of cats and rats. These reactions depend on the activation of trigeminal afferent nerves and release of neuropeptides. A brief stimulation causes vasodilation, which is mainly mediated by calcitonin gene-related peptide (CGRP). A longer stimulation results in plasma extravasation which is mediated mainly by substance P (SP) and prostaglandins in the pulp. In adjacent oral tissues the mechanisms following stimulation or local irritation are more complex and other mediators are also involved. Nitric oxide (NO) which is instantly produced in the tissues is such a novel mediator. The chemosensitive nature of the nerves involved (capsaicin sensitive) may lead to their activation also by inflammatory mediators released in the tissues. Thus, sensory nerves may modulate the progress of inflammation. Since sensory nerves in oral tissues are often the first structures to be activated during clinical procedures, tissue reactions that occur can be assumed to be initiated and perpetuated by the sensory neuropeptides. Much work is now made to modulate the sensory nerveinduced cascade of events in oral tissues to find new treatment strategies. PMID- 9729667 TI - [Intra-arterial administration of Dalacin C phosphate (clindamycin) in the therapy of osteomyelitis of the jaws]. AB - In 8 cases of maxillary and mandibular osteomyelitis which had failed to respond to traditional surgical and medicinal methods of treatment, a significant improvement could be achieved (in 7 of the 8 cases) by applying an intra-arterial clindamycin therapy. Treatment was carried out similar to the intra-arterial cytostatic perfusion treatment of tumors in the head and neck areas. Clindamycin was administered in daily dosages of 900 mg over a period of 8-23 days. PMID- 9729668 TI - [What is beauty?]. AB - The face is a most important part of our features because of the importance it plays in communication. This paper looks at a means of defining what is normal and also what is beautiful. The face is a three-dimensional object and therefore needs to be measured in three dimensions. This paper describes an optical surface scanning system which has been used to analyze the faces of 40 normal patients with a Class I occlusion and also the faces of 30 models from the No. 1 Agency in London. The scans of each group has been separated into males and females, and the faces averaged to give an average normal male face and an average normal female face, an average model male face and an average model female face. These faces have then been compared by a special registration programme which enables the observer to compare the faces in three dimensions and record the amount of difference between them. The value of the system in forensic science for the identification of profiles in photographs is also illustrated as is its use in the building up of the soft tissues over the skull. The importance of which parts of the face are genetically determined has also been looked at by special shape analysis programmes and these are described. PMID- 9729669 TI - [Pathogenesis of apical periodontitis and its effects on the body]. AB - During the last 25 years there have been major advances in understanding the etiology, pathogenesis and maintenance of inflammatory processes taking place in the periapical space. Polymicrobial infection of the pulp chamber is of primary importance in initiating periapical inflammation. Egress of bacteria and their antigenes stimulate the immune system to form a granulation tissue around the apical area. Local immune response eliminates excess number of invading organisms. However, in parallel with protective reactions, local activity of immunocompetent cells and their soluble products also contribute to tissue damage, bone resorption and perpetuation of inflammation. Present data indicate that interaction of T-lymphocytes and macrophages is crucial in this process. PMID- 9729670 TI - [Caries, fluorosis and salt fluoridation in the city of Szeged]. AB - Salt fluoridation is effective at inhibiting caries, but fluorosis prevalence data are different. The purpose was to undertake a blind caries and tooth mottling study of 14-yrs-olds from S. E. Hungary who did (Test) or did not (Control) live, until 1985, in a 350 pp F-/kg salt fluoridated area during their early years of life. In Szeged, blind clinical and anterior tooth-mottling, radiographic and photographic recording of 49 previously salt-fluoridated and 59 non-salt-fluoridated subjects were undertaken by one examiner. In Glasgow, four dental and two lay staff scored the projected 35 mm colour transparencies of each pupil's teeth (13-23), for tooth mottling. Mean DMFS scores were 9.18 (SD = 10.72) and 4.51 (SD = 6.4) for Test and Control users respectively (p < 0.01). Clinically three Test children had fluorosis of 10 teeth, with 8 teeth in two Controls. In a sole Test case was "fluorosis" photographic unanimity obtained, and non-unanimous "possible fluorosis" was recorded by two-four 'jurors' for only three other Test and two Control subjects. No evidence was found that significant fluorosis resulted in subjects exposed previously to 350 ppm F-/kg salt early in life, but no caries benefit was demonstrated after the 11.5-yr salt fluoridation gap. These data emphasize (a) the superiority of substained community-delivered fluoridation, and (b) the need to maintain constant fluoride delivery to tooth surfaces, and certainly well beyond 10 years of age. PMID- 9729671 TI - [Microbial study of the surface of malignant tumors of the oral cavity]. AB - Malodour and other infection sequelae may increase the morbidity, compromise the oral well-being of the patient suffering from maxillo-facial neoplasia. The purpose of our study was to evaluate the differences of the oral bacterial flora attached to the tumor and the contra lateral sound surfaces. Swabs were obtained and samples were microbiologically cultured. It was concluded, that carcinoma surface biofilm harbors increased levels of both aerobs and anaerobs. PMID- 9729673 TI - [Chinolone--the survey remembered]. PMID- 9729674 TI - [Pruritus. Causes and treatment possibilities]. PMID- 9729672 TI - [Comparative clinical study of amalgam fillings]. AB - Two different types of amalgam alloys (Strafill NG2, [Ogussa] and googfill 700 [Ogussa]) were placed into 106 contralateral, homologous molar tooth-pairs of 81 patients. Scoring after two years showed significant difference in case of marginal ditching favouring the non gamma2 alloy. However significant differences were not found in case of other observed failures, such as bulk fracture, surface roughness and secondary caries. PMID- 9729675 TI - [Treatment of hypertension. Recommendations of the Hypertension Society]. PMID- 9729676 TI - [Alzheimer disease. Diagnosis and treatment]. PMID- 9729677 TI - [Practical considerations of immunoprophylaxis in viral hepatitis]. AB - Hepatitis virus infections belong to the major etiological factors of both acute and chronic liver diseases. During the last years--apart from the passive immunoprophylaxis through the administration of immunoglobulin--safe and efficacious hepatitis vaccines (against hepatitis A and B viruses) has also became available in Hungary. In this review the authors focus on the practical considerations of the immunoprophylaxis of hepatitis viruses. General consensus concerning the clinical use of hepatitis A vaccine has not been accepted, and human immunoglobulin is the only acceptable measure to prevent postexposure hepatitis A virus infection. Hepatitis B immunoglobulin (HBIG) is administered to prevent hepatitis B infection after exposure of HBV-contaminated body fluids and in infants born to HbsAg-positive mothers. Recombinant hepatitis B vaccine has the crucial note in the WHO's program for eradication of hepatitis B. While in the areas of high endemicity the implementation of universal hepatitis B vaccination is recommended, in the countries with low HbsAg-carrier prevalence infants' and/or adolescents' vaccination can offer alternative choices. Prevention of health-care workers against hepatitis virus infection and the vaccination of patients with chronic liver disease are also of great importance. There is also an urgent need to establish multidisciplinary professional cooperation to be able to carry out effectively the WHO's recommendations for prevention of hepatitis B infection. PMID- 9729678 TI - [The role of ultrasonography and biopsy following kidney transplantation]. AB - Several complications can occur during both the early and late postoperative periods after kidney transplantation. The methods used to follow up 575 kidney transplanted patients, (transplantations performed between October 1979 and November 1997) in the early (within 6 weeks) and late postoperative periods have been assessed. The diagnostic value of core biopsies and ultrasound examinations, the prevalence of complications, and the applicability of the diagnostic tools in the evaluation of the graft status and viability were analyzed. In the early postoperative period, graft rupture occurred more frequently after biopsy than in the late period (7.4% vs 0.82%), this leading graft loss in 18 of 20 cases. The sonographically diagnosed morphologic and functional changes were also analyzed. Sonography proved a very accurate method for the detection of perirenal fluid collections and masses and severe vascular complications. The data demonstrated that biopsy is indicated in the early postoperative period when the result of sonography is doubtful. In the late postoperative period, biopsy should be performed in every case. PMID- 9729679 TI - [The in vitro effect of recombinant human growth hormone on lymphocyte and granulocyte function in healthy and uremic children]. AB - The aim of the study was to establish the effect of GH on immune functions in 22 healthy and in 11 uremic children, in vitro. Oxydative burst of granulocyte in the presence of GH measured by chemiluminescence and lymphoblast proliferation to GH and lectin stimuli were studied. Gene expression of GH receptor was analyzed with reverse transcriptase polymerase chain reaction (RT-PCR) method. The metabolic burst of granulocytes individually differed, specially in the chronic renal failure (CRF) group (60%) showed rather dose and time-dependent increase, the GH had only a priming effect. In 59% of the healthy children the GH stimulated the lympho-proliferative response itself or interaction with the lectin (ANOVA-test) and increased the spontaneous lymphoproliferation in 45% of the uremic patients. The GH receptor mRNA expression differed in the childrens lymphocytes, showing no correlation with the effect of GH on lymphoproliferation. The GH has a cytokine-like role in the regulation of the human immune system, and the GH treatment in uremic children is rather stimulating on immune functions. PMID- 9729680 TI - [Vestibular control of the cardiovascular system]. AB - Due to modern living conditions the human cardiovascular system is frequently exposed to rapid or/and prolonged changes in gravitational forces. These transients are very short compared to the timescale of the evolution causing substantial difficulties in adaptation. As it has been many times proven experimentally since the first observation in 1922, the vestibular system affects directly the regulation of the cardiovascular system. For example, bilateral transsection of the vestibular nerve of cats significantly disturbs the compensation of acute hypotension induced by lowering the animal's head. The results of human studies also indicate the existence of vestibulo-sympathetic reflexes. Vestibular excitation caused by caloric to other stimuli results in increased sympathetic efferent activity. Several groups or nuclei in the brain stem (medial vestibular ncl., subretrofacial ncl., the lateral area of tegmentum) were confirmed to have important mediatory function in the central organization of the vestibulo-sympathetic reflex. However, the role of vestibular system in long-term adaptive responses of the vascular system of chronic changes in body position is not classified yet. Such a possible role is suggested by our experimental findings during the last decade. Electronmicroscopic examinations indicated that two-week long orthostatic load due to head-up tilting changes significantly and differently the innervation density of blood vessels in the extremities of rats. There also have been a significant amplification of acute myogenic response to intraluminar pressure-elevation in the saphenous vein. We suppose that adrenergic mechanisms under vestibular control are at least in part responsible for the regionally different adaptive changes including structural remodelling. Spectral analysis of the arterial blood pressure suggested that a two week-long orthostatic load can already alter the overall control of the cardiovascular system in rats. PMID- 9729681 TI - [Successful treatment of acute ethylene glycol poisoning with plasmapheresis]. AB - The authors show the differential diagnostics, therapy and patomechanism of the ethylene glycol intoxication in connection with review of their young patient became acute paraparetical in consequence of the intoxication. In the therapy of the ethylene glycol intoxication, instead of the traditionally proved hemodialysis their patients was treated with great efficiency with plasmapheresis. The authors show the patomechanism of a patient came with atypical Guillain-Barre-Syndrome. In the hinterground of an acute paraparesis was an ethylene glycol intoxication identified. In the therapy of this intoxication their patients was treated with great efficiency with plasmapheresis instead of the traditionally proved haemodialysis. They touch upon the possibility of monitoring with gas chromatography of the patients plasmapheretised, and call attention the easy check of the sodium fluorescein content of the antifreeze additive therefore the rapid recognition of the intoxicated status. PMID- 9729682 TI - The use of psychotropic medication in preschoolers: some recent developments. AB - OBJECTIVE: To provide an overview of the use of psychotropic drugs in preschoolers. METHOD: Literature review. RESULTS: Although controversy persists, the evidence suggests that preschool children are being given an increasing number of psychotropic drugs, especially by general practitioners and pediatricians. CONCLUSION: There is an urgent need to formally evaluate the efficacy of psychotropic medication for young children. PMID- 9729683 TI - The use of psychotropic medication in preschoolers: indications, safety, and efficacy. AB - OBJECTIVE: To review the indications, safety and efficacy of psychotropic medications used in preschoolers. METHODS: Proprietary prescription-use databases indicate that practitioners are prescribing psychotropic medications for preschool patients at an increasing rate. A Medline search was conducted using drug exposure for children below the age of 6 years to identify efficacy and safety reports of these agents in the preschool age-group. RESULTS: The search yielded 22 reports that mention exposure to medications, including maternal exposure, accidental overdose, and adverse events in preschool children. Safety issues highlight the age-specific vulnerabilities of this age-group, including hepatotoxicity from valproic acid, among others. In addition, the prominence of adverse-event responses in this age group may be related to polypharmacy not seen in school-age children or adolescents. Less than a dozen controlled efficacy studies of psychotropic agents were identified for children in the preschool age group. These are limited by the small numbers of subjects in the reports. Only 2 disorders described in the Diagnostic and Statistical Manual of Mental Disorder (DSM-IV), attention-deficit hyperactivity disorder (ADHD) and autistic disorder, are mentioned. The Food and Drug Administration (FDA) approved psychotropic medications for preschoolers but limited their use to medical purposes, not psychiatric, with the exception of use for ADHD. CONCLUSION: Because data about psychotropic drug safety and efficacy in adults have not been extended to children, new psychopharmacological research is required before clinicians can use these agents to treat psychiatric disorders in the preschool age-group. PMID- 9729684 TI - Pediatric psychopharmacology and the interaction between drugs and the developing brain. AB - With increasing frequency, psychotropic medications are being prescribed to young children, often for long periods of time. The interaction between psychotropics and the developing brain has not been systematically investigated in humans. Data collected from animals suggest that developing neurotransmitter systems can be exquisitely sensitive to early inhibition or stimulation by pharmacological agents, which can lead to permanent changes in adult life. Most of these data are collected from rodents, and their extrapolation to humans is difficult. More relevant models could be developed for instance using primates. In humans, the focus of research has traditionally been on the possible teratogenic effects of prenatal drug exposure. Recently introduced quantitative imaging techniques can offer new approaches to studying the effects of psychotropics on the developing brain. This research has clear implications for the safety and efficacy of psychopharmacologic drug in children. PMID- 9729685 TI - Ethical and pragmatic issues in the use of psychotropic agents in young children. AB - OBJECTIVE: To provide an overview of the knowledge base concerning the prescribing of psychotropic agents in young children with mental disorder and related mental health problems. METHOD: Relevant information is reviewed concerning the knowledge base available to inform pediatric psychopharmacology prescribing practices. RESULTS: Very few psychoactive medications have been adequately tested for safety and efficacy in young children, despite relatively high rates of prescribing. CONCLUSION: Behavioral and psychotherapeutic strategies are often the wisest first therapeutic intervention for this age group. Psychotropic medications may be required, but should be used cautiously in young children, while additional studies are being conducted. PMID- 9729686 TI - Autism and other pervasive developmental disorders: exploring the dimensional view. AB - OBJECTIVE: To examine empirical data on children with autistic disorder (AD), Asperger's disorder, and pervasive developmental disorder not otherwise specified (PDD-NOS) for continuities or distinguishing features between disorder and to see to what extent the Diagnostic and Statistical Manual of Mental Disorder (DSM-IV) diagnostic criteria-reflect observed data. METHOD: Studies were identified in 4 ways. 1)A Medline search from 1976 to the present of articles with the key words autism, pervasive developmental disorder, autistic spectrum disorder, and Asperger; of these articles, those with mesh headings or textwords "cluster," which identified cluster analyses deriving pervasive developmental disorder (PDD) subtypes, were retained 2) The Journal of Autistic and Developmental Disorder from 1990 to the present was hand-searched to identify other empirically derived studies on diagnosis, prevalence, classification, and validity of PDD subtypes. 3) Key review articles were searched for their references. 4) The references of all identified articles were searched. RESULTS: Eight cluster studies were retained for their relevance to diagnostic issues, as were 7 empirically derived studies delineating clinic characteristics of children will AD, Asperger's syndrome, or PDD-NOS. Data suggests that children with PDD may fit into 1 of 2 overlapping groups, including a lower-functioning group with greater developmental compromise, social aloofness, and a greater number of autistic symptoms and a higher-functioning group with higher IQ, fewer autistic symptoms, and more prosocial behavior. The PDD subtype resemble each other and can be seen as existing o a continuum, differing only by degree of impairment. CONCLUSION: Children exhibiting the triad of autistic impairments can be seen as suffering from disorders on a PDD continuum. While the DSM-IV does identify a lower functioning autistic group (AD), the higher-functioning group is less well served. Asperger's disorder as defined in the DSM-IV is not clearly distinguishable from AD and PDD-NOS, and the PDD-NOS subcategory is not operationalized. Further research is required to elaborate criteria for the higher-functioning PDD group, and measures related to etiology, outcome, and treatment response may help determine which diagnostic criteria can meaningfully separate one disorder from another. PMID- 9729687 TI - Typical and atypical antipsychotics in adolescent schizophrenia: efficacy, tolerability, and differential sensitivity to extrapyramidal symptoms. AB - OBJECTIVE: To review the existing literature on the efficacy and tolerability of antipsychotics for adolescent psychosis. The review focuses in particular on literature regarding adverse effects that are thought to have an increased incidence in young patients and on the possible neurobiological bases for such differential sensitivity. METHOD: Pertinent studies were sought using Medline searches, supplemented by selected bibliographies, and reviewed. RESULTS: There is a relative paucity of research in this area; in particular, well-controlled trails are lacking. The existing literature suggests fairly good efficacy of both typical and atypical antipsychotics in the treatment of psychotic disorders in children and adolescents. However, the incidence of certain side effects, particularly extrapyramidal symptoms (EPS). is found to be higher in younger patients compared with adults. Positron emission tomography (PET) receptor studies in adults have demonstrated that the incidence of EPS is related to dose dependent dopamine type-2 (D2) receptor occupancy and that there is a significant relationship between the number of these receptors and age. CONCLUSIONS: Improved tolerability is leading to the increasing us of atypical antipsychotics for adolescent patients, though these new drugs to have specific adverse effects of their own. There is a need for more controlled studies of atypical antipsychotics in children and adolescents. In particular, dose-finding studies are needed to determine the optimal dose range to produce the greatest improvement with the least side effects for each of these drugs. PMID- 9729689 TI - The integration of child psychiatry into a psychiatry clerkship OSCE. AB - OBJECTIVE: To integrate child psychiatry into a psychiatry clerkship OBJECTIVE Structured Clinical Examination (OSCE). METHOD: Child psychiatry OSCE stations were designed to evaluate clerk' skills in the identification of 4 common conditions. Child psychiatrists wrote case scenarios and checklists and supported standardized patient (SP) training for the stations. A bank of 4 child psychiatry OSCE stations is now available for use in the psychiatry OSCE. Child psychiatry faculty have been trained as examiners for ongoing administration of his OSCE. RESULTS: This bank of child psychiatry OSCE stations has examined 402 clerks. Mean student scores for content were 68% to 86% and for process were 69% to 76%. Station reliability and examiner feedback were acceptable. CONCLUSIONS: Child psychiatry has been successfully integrated into a psychiatry clerkship OSCE. Although the commitment in terms of monetary and faculty costs has been considerable, the accompanying educational benefits of such integration warranted this expense. PMID- 9729688 TI - What does early antisocial behaviour predict? A follow-up of 4- and 5-year-olds from the Ontario Child Health Study. AB - OBJECTIVE: To examine the predictive accuracy of antisocial behaviours among 4- and 5-year-old children for problem behaviours 4 years later (ages 8 and 9 years). METHOD: Data from the Ontario Child Health Study (1983) and Follow-up (1987) are used. Predictive accuracy is conceptualized using positive predictive value (PPV) and sensitivity. The predictive accuracy of early antisocial behaviors for the 1987 outcomes is examined overall, by gender, by variable thresholds of predictor and outcome be gender, and by using contextual variables alone or in combination with antisocial behaviour recorded in 1983. Resulting: The predictive accuracy of 1983 antisocial behaviour for 1987 outcome is generally modest and differs by gender (better for boys for externalizing disorder [PPV = 41%, sensitivity = 57%]; better for girls for internalizing disorder [PPV = 13%, sensitivity = 80%]; better for boys for conduct problems [PPV = 54%, sensitivity = 21%]?. Using either gender-specific thresholds or gender-neutral thresholds does not alter predictive accuracy in a consistent way, nor does the use of a single contextual variable. Use of a cumulative risk index increases PPV but decreases sensitivity. CONCLUSION: The predictive accuracy of antisocial behaviour in 4-and 5-years-old children over 4 years in a nonclinical community population is limited. The clinical, research, and policy implications of this work are discussed. PMID- 9729690 TI - 47,XYY karyotypes and pervasive developmental disorders. AB - OBJECTIVE: The presence of a 47, XYY karyotype in boys with pervasive developmental disorders (PDDs) has rarely been described in the past. Herein, 2 boys with PDDs and a supernumerary Y chromosome are presented. METHODS: The case histories of the 2 patients are described along with the results of associated testing. The literature on psychosocial development as well as brain morphology and physiology in males with 47, XYY karyotypes is reviewed. RESULTS: Both boys had presentations typical of PDDs, one with autistic disorder and the other with PDD not otherwise specified. CONCLUSION: The finding that, in a clinic for children with developmental disorders, 2 of 40 male referrals had 47, XYY karyotypes suggests that the rate of this sex chromosome anomaly may be increased in PDDs. An extra Y chromosome may be related to abnormal brain development, which may, in turn, predispose vulnerable males to PDDs. PMID- 9729691 TI - Attention-deficit hyperactivity disorder subtypes and comorbid disruptive behaviour disorders in a child and adolescent mental health clinic. AB - OBJECTIVE: To assess demographic characteristics and patterns of comorbid disruptive behavior disorders (oppositional defiant disorder [ODD] or conduct disorder [CD]) in subtypes of attention-deficit hyperactivity disorder (ADHD). METHOD: One hundred youths consecutively referred to a community child and adolescent mental health clinic and subsequently diagnosed with ADHD by the Diagnostic and Statistical Manual of Mental Disorders (DSM-IV) criteria were evaluated. The diagnosis was made by a child psychiatrist and was based on information from physicians, parents, teachers, and diagnostic interviews with the youth and their parents. RESULTS: The major findings were: 1) ADHD combined (C) type was diagnosed in 78% of the subjects, while 15% had inattentive (1) type and 7% had hyperactive-impulsive (HI) type; and 2) patterns of comorbid disruptive behavioural disorders significantly differed among subtypes. Specifically, subjects with the I type showed lower rates of comorbid ODD than those with the C type (33% and 85%; P < 0.001) and HI type (33% and 100%; P = 0.005); subjects with the HI type displayed a higher prevalence of CD than those with the I type (57% and 0%; P = 0.005) and C type (57% and 8%; P = 0.003). These results should be considered tentative because the reliability of the diagnostic procedures was not formally assessed and the number of subjects in the I and HI groups was small. CONCLUSION: ADHD subtypes showed significant differences in the distribution of comorbid disruptive behaviour disorders. These results support the utility of ADHD subtypes but should be replicated with a larger sample of I and HI type subjects using more rigorous diagnostic methods. PMID- 9729692 TI - Adolescents on neuroleptic medication: is this population at risk for tardive dyskinesia? AB - OBJECTIVE: The assess the incidence of tardive dyskinesia (TD) in a sample of adolescents treated with neuroleptic medication and to identify the presence of any risk factors for TD within the affected group. METHOD: A retrospective chart review was conducted for 40 cases. The Abnormal Involuntary Movement Scale (AIMS) was used to measure side effects from medication at 6-month intervals over 2 years. Drug exposure was converted to chlorpromazine (CPZ) equivalent and the presence of risk factory for TD, such as a diagnosis of affective disorder, medication noncompliance, early age of illness onset, and concomitant antiparkinsonian medication, was also noted. RESULTS: Of the 40 cases reviewed, 2 patients (5%) met diagnostic criteria for TD, and another 5 patients (12.5%) showed symptoms of TD. CONCLUSIONS: TD is a serious risk at any age. Medication noncompliance, early age of illness onset, and concomitant use of antiparkinsonian medication may increase susceptibility to TD and should be carefully monitored. PMID- 9729694 TI - Hospital separations for delirium in Canadian psychiatric patients from 1985 to 1994. PMID- 9729693 TI - Culture and Munchausen-by-proxy syndrome: the case of an 11-year-old boy presenting with hyperactivity. AB - OBJECTIVES: To discuss some of the challenges presented to the clinician who deals with a possible Munchausen-by-proxy (MBP) syndrome. METHOD: The case of an 11-year-old boy presenting with hyperactivity is discussed. Information from the initial assessment and the 9-month follow-up period is presented. We highlight some cultural considerations as they apply to this immigrant family. A commentary by Dr H Schrier follows the presentation. RESULTS: The positive outcome is discussed in relation to the validation of the diagnosis as well as to cultural issues. CONCLUSION: Cultural issues and dynamic factors may be important when we consider the diagnosis of MBP syndrome in an immigrant family with different expectations from our health care system. PMID- 9729695 TI - Bupropion-SR-induced increased libido and spontaneous orgasm. PMID- 9729696 TI - Provocation of obsessive-compulsive behaviour and tremor by olanzapine. PMID- 9729697 TI - The serotonin syndrome as a learning opportunity. PMID- 9729698 TI - Beneficial effects of combined L-dopa and central anticholinergic in a patient with severe drug-induced parkinsonism and tardive dystonia. PMID- 9729699 TI - Bipolar II depressed outpatients seek treatment more quickly than do unipolar outpatients. PMID- 9729700 TI - Congenital anomalies of the hand. AB - This manuscript reviews the basic and most common types of congenital hand anomalies seen by the pediatrician. Each disorder is categorized into the standard classification system used, and the main highlights of each condition are detailed with representative illustrations. Other system anomalies associated with congenital hand problems are also identified. PMID- 9729701 TI - Developmental implications of head growth following intracranial hemorrhage. AB - This study examined the association between head size at birth, discharge, and 1 year and developmental outcome at 1 year in preterm infants, with and without intracranial hemorrhages (ICH) or associated periventricular echodensities (PVE). The data indicated that most sick preterm infants with small heads at discharge achieved appropriate head sizes at 1 year. Analyses of the 1-year mental and motor performances of 125 subjects revealed that for subjects who did not develop ICH, appropriate head sizes at birth and discharge were associated with good developmental outcome, whereas infants with small heads (< two standard deviations below the mean for age) before hospital discharge were more likely to show poorer developmental outcome at 1 year. For subjects with ICH, birth and discharge head circumference were not predictive of 1-year developmental status; however, normal head size at 1 year was associated with better outcome. This was true for children with transient PVE as well. However, persistent periventricular echodensities were associated with both mental and motor deficits at 1 year, regardless of head growth. PMID- 9729702 TI - Rabies prophylaxis following the feeding of a rabid pony. AB - A survey was performed to identify people who were exposed to a rabid pony and determine whether or not they received rabies postexposure prophylaxis (PEP). Sixty-one visitors who came in contact with the rabid pony were identified. These visitors heard about the rabid pony via the news media. Forty-five visitors were exposed during the 2 weeks before the pony died. Thirty-two of these 45 visitors received PEP. Thirty-one visitors had or may have had saliva contact to an open wound or mucosa and all 31 received PEP. Fourteen visitors had no saliva contact to a wound or mucosa and one received PEP. Sixteen visitors were exposed before the pony was potentially shedding rabies virus and one received PEP. No visitor was bitten by the pony. Most of the persons 31/33 (94%) who received PEP had an exposure for which PEP was indicated. Nonbite transmission of rabies is discussed. PMID- 9729703 TI - Hydrophobic horse sense. PMID- 9729704 TI - Use of glycopyrrolate and other anticholinergic medications for sialorrhea in children with cerebral palsy. AB - Fifty-four parents/caretakers of children with cerebral palsy were surveyed regarding their use of antisialorrheic medication for excessive drooling. Glycopyrrolate was used by 37 of 41 respondents, with significant improvement in drooling noted in the vast majority (95%) of cases as indicated by a five-point rating scale. Side effects (dry mouth, thick secretions, urinary retention, or flushing) surfaced in almost half (44%) of the patients but necessitated discontinuation of pharmacologic treatment in less than a third. While larger clinical studies are needed, our preliminary data indicate a trial of glycopyrrolate should be considered in children with cerebral palsy where drooling is a significant problem. PMID- 9729706 TI - Munchausen syndrome by proxy. AB - Munchausen syndrome by proxy is the most difficult form of child abuse. It carries substantial morbidity and mortality. The diagnosis relies on appropriate suspicion and careful investigation. The psychological illness/need of the perpetrator is the main clinical feature. Early recognition and appropriate intervention prevent further abuse and criminal actions. PMID- 9729707 TI - Treatment of pulmonary hypertension with inhaled nitric oxide during hepatic transplantation in an adolescent: reversibility of pulmonary hypertension after transplantation. PMID- 9729705 TI - Diagnosis and treatment of an adolescent with comorbid type 1 diabetes mellitus and anorexia nervosa. AB - The authors describe the successful treatment of a 17-year-old female with comorbid type 1 diabetes mellitus and anorexia nervosa within the context of a residential program in a tertiary care facility. Assessment and treatment of the complex combinations of the psychological and medical symptoms involved in this patient required the interaction of medical, psychological, and nutritional services. Diagnostic and treatment challenges are discussed. PMID- 9729708 TI - Dubin-Johnson syndrome as a cause of neonatal jaundice: the importance of coproporphyrins investigation. PMID- 9729709 TI - Nonambulatory "pedestrians": infants injured by motor vehicles in driveways. PMID- 9729710 TI - Pattern formation and developmental mechanisms. PMID- 9729711 TI - Pattern formation and developmental mechanisms. Converging views of diverging pathways. PMID- 9729712 TI - Microbial asymmetric cell division: localization of cell fate determinants. AB - The genetic mechanisms that control asymmetric cell divisions--yielding progeny cells that differ from one another--have been conserved among prokaryotes, eukaryotic microbes, and higher organisms. All use the paradigm of regulatory protein localization as a way of translating genetic information into three dimensional space. PMID- 9729713 TI - Asymmetric cell division: lessons from flies and worms. AB - Insights into the mechanisms of asymmetric cell division have recently been obtained from studies in genetically amenable systems such as Drosophila and Caenorhabditis elegans. These studies have emphasized the importance of cortically localized polarity organizing molecules, adapter molecules, and the actin cytoskeleton in controlling unequal segregation of cell-fate determinants and spindle orientation. The control of asymmetric cell divisions by Wnt signaling in C. elegans and Frizzled signaling in Drosophila reveals additional mechanisms for modulating cellular polarity and suggests that there are some similarities between the two systems. PMID- 9729714 TI - Common threads in eukaryotic circadian systems. AB - Within the past 18 months, common regulatory patterns have emerged among eukaryotic circadian systems--extending from fungi through to mammals. Heterodimeric complexes of PAS-domain-containing transcription factors play positive roles in clock-associated feedback loops, and classic clock proteins like FREQUENCY (FRQ), PERIOD (PER), and TIMELESS (TIM) appear as negative elements. Post-transcriptional control governs the amount and type of FRQ and makes the clock responsive to temperature. PMID- 9729715 TI - Complexity of EGF receptor signalling revealed in Drosophila. AB - The multiple roles of the Drosophila epidermal growth factor receptor (EGFR) require that its activation is regulated precisely. Recent work has highlighted two important control mechanisms: the existence of multiple ligands with distinct properties and the interaction between EGFR pathway and other signalling pathways. The integration of signalling pathways into networks is beginning to be understood. PMID- 9729716 TI - Ras--a versatile cellular switch. AB - With the number of known roles played by Ras proteins increasing rapidly, finding answers to how the diverse cellular responses are triggered is becoming increasingly pertinent. Although our understanding of the control of specificity of signal transduction is still small, the combination of biochemical, structural and genetic analyses is starting to reveal how the cell-specific responses to Ras activation are controlled. PMID- 9729717 TI - Specificity in signaling pathways: assembly into multimolecular signaling complexes. AB - A critical issue in the field of signal transduction is how signaling molecules are organized into different pathways within the same cell. The importance of assembling signaling molecules into architecturally defined complexes is emerging as an essential cellular strategy to ensure specificity and selectivity of signaling. Scaffold proteins function as the pillars of these transduction complexes, bringing together a diversity of signaling components into defined ultramicrodomains of signaling. PMID- 9729718 TI - Hox proteins meet more partners. AB - The Hox genes are clustered sets of homeobox-containing genes that play a central role in animal development. Recent genetic and molecular data suggest that Hox proteins interact with pre-existing homeodomain protein complexes. These complexes may help to regulate Hox activity and Hox specificity, and help cells to interpret signaling cascades during development. PMID- 9729719 TI - Propagation and localization of Wnt signaling. AB - Cellular mechanisms for the transport and localization of Wnt signaling components are important for the propagation, distribution, and polarization of Wnt signals in embryonic tissues. Wnt signals are distributed through tissues by vesicular transport of Wnt proteins, localized in embryos by directed transport of cytoplasmic Wnt-signaling components, and propagated asymmetrically during cell division. PMID- 9729720 TI - Notch signaling: direct or what? AB - Notch signaling has been implicated in a wide variety of processes from cell-fate decisions, tissue patterning and morphogenesis to human diseases and cancer. A model for Notch directly regulating gene expression has been proposed and at least two signaling pathways have been identified; however, the molecular mechanism(s) by which Notch signaling produces so many outcomes remains unclear. PMID- 9729721 TI - BMPs, Smads and metalloproteases: extracellular and intracellular modes of negative regulation. AB - Bone morphogenetic (BMPs), members of the TGF-beta superfamily, have critical functions in many biological contexts. Recent findings in Drosophila and vertebrates suggest that BMP signaling can be modulated extracellularly and intracellularly by the availability of BMP inhibitors and Smads, respectively. PMID- 9729722 TI - New players and puzzles in the Hedgehog signaling pathway. AB - Members of the Hedgehog (Hh) family of signaling proteins control cell fates and proliferation during animal development in part by regulating the transcription of specific genes. Depending on the tissue, Hh can act over long or short distances, to signal directly or by inducing secondary signals. Recent discoveries include new components of the pathway as well as novel regulatory mechanisms involving cholesterol, proteolysis, and the cytoskeleton. The role of Hh in carcinogenesis is underscored by the identification of mutations in several pathway components in skin and brain tumors. PMID- 9729723 TI - Ascidian embryogenesis and the origins of the chordate body plan. AB - For more than a century, ascidians have been a widely used system for classic embryological studies. Ascidians possess simple, well-defined cell-lineages, compact genomes, rapid development and world-wide distribution. Transgenic DNA can be introduced into developing embryos using simple electroporation methods. The ascidian larva represents the most simplified chordate body plan and provides a useful model for studying the molecular pathways underlying the morphogenesis and differentiation of the notochord and neural tube. PMID- 9729724 TI - The zebrafish organizer. AB - Signals from the organizer play a crucial role in patterning the vertebrate embryo. Recent molecular analysis of zebrafish mutations has established an essential role for BMP2 and chordin in organizer function and has identified one eyed pinhead as a novel EGF-like gene involved in prechordal plate and endoderm formation. In addition, embryological studies have suggested that the zebrafish organizer is induced by extraembryonic cues and have defined two novel organizing centers that pattern the nervous system along the anterior-posterior axes. PMID- 9729725 TI - The maternal-to-zygotic transition in embryonic patterning of Caenorhabditis elegans. AB - Maternal factors laid down in the oocyte regulate blastomere identities in the early Caenorhabditis elegans embryo by activating zygotic patterning genes and restricting their expression to the appropriate lineages. A number of early acting zygotic genes that specify various cell fates have been identified recently and their temporal and spatial regulation by maternal factors has begun to be elucidated. PMID- 9729727 TI - Somitogenesis: segmenting a vertebrate. AB - The partitioning of the vertebrate body into a repetitive series of segments, or somites, requires the spatially and temporally co-ordinated behaviour of mesodermal cells. To date, it remains unknown how applicable our knowledge of the genetic mechanisms governing Drosophila segmentation will be to that of vertebrates, though recent results indicate some degree of conservation. Genetic studies in the mouse point to a major role for the Notch-Delta signalling pathway in somite formation. Furthermore, a molecular clock may be 'ticking' in the presomitic mesoderm. PMID- 9729726 TI - Epithelial/mesenchymal interactions and branching morphogenesis of the lung. AB - The establishment of branched tubular epithelial structures is critical for the viability of multicellular organisms: the tracheal system in Drosophila and the vertebrate lung being two such structures. Although there are obvious differences in the complexity of these branched organs, many of the underlying mechanisms and genes regulating their development appear to have been evolutionarily conserved. PMID- 9729728 TI - Cbfa1 in bone development. AB - A factor fundamental to bone formation has been identified. Gene targeting shows that core-binding factor alpha 1 (Cbfa1) plays an essential role in bone formation and osteoblast differentiation. Thus, it is now possible to begin examining the molecular mechanism of bone formation--especially osteoblast differentiation. PMID- 9729729 TI - Lipids engineering and design membrane proteins. Web alert. PMID- 9729730 TI - Potpourri: effects of unsaturation on lipid structure; plasma cholesterol ester and lipid-transfer proteins; and cholesterol-sensing proteins and cellular cholesterol movement. PMID- 9729731 TI - Diversity in the fatty-acid conformation and chain packing of cis-unsaturated lipids. AB - Recent X-ray diffraction and Fourier transform IR spectroscopic studies have unveiled a great diversity in the molecular conformation and chain packing of lipid molecules containing cis-unsaturated fatty-acid chains. Specifically, a dramatic diversity in the olefinic conformation, subcell packing and chain-chain interactions has been clearly revealed by crystal structures of principal cis monounsaturated fatty acids. The structural diversity is most manifest in oleic acid crystals. These findings were applied to analyses of the complicated structural transformation of diacylglycerols and triacylglycerols containing oleic acid and saturated acid moieties, in which the stabilization of the oleic acid causes the complexity of the transformation and mixing behavior. Although this knowledge has been obtained in the crystalline state, one may assume that the structural diversity of these unsaturated molecules plays a similar role in the lipids of biomembranes, lipoproteins and lipid deposits in which aliphatic chain packing is a critical problem, since most lipid domains can undergo liquid to solid or solid to liquid alterations. PMID- 9729732 TI - The implications of the structure of the bactericidal/permeability-increasing protein on the lipid-transfer function of the cholesteryl ester transfer protein. AB - The cholesteryl ester transfer protein (CETP) is evolutionarily related to the bactericidal/permeability-increasing protein (BPI). The recently solved structure of BPI shows an elongated, boomerang-shaped molecule, with two hydrophobic pockets opening to its concave side. These pockets each contain a phospholipid molecule. A model of CETP, based on the recently solved crystal structure of BPI, provides the basis for interpreting functional studies on CETP. In this model, C terminal residues 461-476, which were shown to be required for neutral lipid transfer between plasma lipoproteins, from an amphipathic helix covering the opening of the N-terminal pocket. A possible lipid-transfer mechanism for CETP, with the initial step involving the disordering of lipids in the lipoprotein surface, followed by the flipping and entry of a lipid molecule into the hydrophobic lipid-binding pocket, is hypothesized in light of structural evidence and recent studies. PMID- 9729733 TI - Four cholesterol-sensing proteins. AB - What is the connection among the following three medical conditions: Niemann-Pick type C disease (a cause of mental retardation and early death), systemic lipidosis (in which an obscure side effect of numerous drugs transforms lysosomes into lamellar bodies), and holoprosencephaly (a catastrophe in embryonic development)? Recent evidence suggests that the pathogenesis in each use involves impaired sensing of cellular cholesterol. PMID- 9729735 TI - Antibody engineering. AB - Among the most important advances in antibody engineering of this past year is the advent of new tools to study the relationship between protein (including antibody) structure and function. Very rapid large-scale mutational analysis of antibodies is now possible by using in vitro transcription and translation. Ribosome display is a rapidly evolving technology for modifying antibody function that offers several potential advantages over phage display. PMID- 9729734 TI - Engineering and design. Recent adventures in molecular design. PMID- 9729736 TI - Redesigning small molecule-protein interfaces. AB - Several recent reports have described the rational redesign of small molecule protein interfaces, generating modified ligands that interact only with suitably mutated proteins. These studies provide powerful reagents for specifically manipulating engineered proteins inside cells, as well as general insights into the factors underlying the binding affinity and specificity of small molecules in general. Progress this year includes the development of allele-specific inhibitors of Src-family tyrosine kinases and the crystal structure determination of a remodeled FKBP-ligand interface. PMID- 9729737 TI - The design of protein-based catalysts using semisynthetic methods. AB - The combination of site-directed mutagenesis and chemical modification has resulted in the preparation of protein conjugates with new and useful properties. Proteins modified with metal-chelating groups are proving useful for mapping tertiary and quaternary interactions using the technique of affinity cleavage. The attachment of cofactors, including pyridoxal and pyridoxamine, has resulted in the preparation of semisynthetic transaminases that display enzyme-like properties, including enantioselectivity, substrate specificity and reaction-rate acceleration. PMID- 9729739 TI - Computer search algorithms in protein modification and design. AB - The computer-aided design of protein sequences requires efficient search algorithms to handle the enormous combinatorial complexity involved. A variety of different algorithms have now been applied with some success. The choice of algorithm can influence the representation of the problem in several important ways--the discreteness of the configuration, the types of energy terms that can be used and the ability to find the global minimum energy configuration. The use of dead end elimination to design the complete sequence for a small protein motif and the use of genetic and mean-field algorithms to design hydrophobic cores for proteins represent the major themes of the past year. PMID- 9729740 TI - Membrane proteins. Channels, pumps and charge separators. PMID- 9729738 TI - Functionalization of designed folded polypeptides. AB - Recent progress in the design of folded polypeptides has led to new catalysts, peptides that bind metal ions, hemes and cofactors, and folded glycopeptides. The development of techniques for postsynthetic and post-translational functionalization promises to further expand the repertoire of accessible motifs. The demonstration of function in complex sites helps to understand protein structure and function and has important implications for the engineering of new proteins with novel properties. PMID- 9729741 TI - Proton translocation by bacteriorhodopsin and heme-copper oxidases. AB - Proton translocation is the means by which free energy is conserved from oxygen reduction by the respiratory heme-copper oxidases and from sunlight by bacteriorhodopsin. Three-dimensional structures at atomic resolution of both proteins have aided functional studies of the proton translocation mechanism. A comparison reveals common structural and functional features that may be unique to the primary proton pumps in biology. PMID- 9729742 TI - The structure and mechanism of the family of retinal proteins from halophilic archaea. AB - Retinal proteins from halophilic archaea provide a unique opportunity to analyze vectorial ion translocation. Studies on its structure, conformational changes, proton conduction and electrogenic steps have helped to elucidate the catalytic cycle of bacteriorhodopsin in increasing detail. Experimental modulation of the vectoriality and ion specificity by altering the substrate availability, point mutations and light conditions for the different retinal proteins allows the proposal of a general model of ion transport for this protein family. PMID- 9729743 TI - Structure and function of the cytochrome bc1 complex of mitochondria and photosynthetic bacteria. AB - Progress has recently been made in the understanding of the function of the cytochrome bc1 complex and related proteins in the context of recent structural information. The structures support many features that were predicted from sequence analysis and biophysical studies, but contain some surprises. Most dramatically, it is apparent that the iron-sulfur protein can take up different positions in different crystals, suggesting a novel mechanism for electron transfer through domain movement. Evidence from studies of mutant strains, in which the function of the sites or the binding of inhibitors is perturbed, has provided clues about the mechanism. PMID- 9729744 TI - Structure of the P-type ATPases. AB - Electron cryocrystallography of precipitant-induced two-dimensional surface crystals of the neurospora plasma membrane H+ - ATPase and tubular crystals of the sarcoplasmic reticulum Ca(2+)-ATPase has recently yielded structure maps for these ion transporters at a resolution of about 8 A. The membrane-embedded regions of these closely related enzymes are similar, but the cytoplasmic regions appear to be significantly different. PMID- 9729745 TI - Synthesis, assembly and structure of gap junction intercellular channels. AB - Gap junction membrane channels assemble as dodecameric complexes, in which a hexameric hemichannel (connexon) in one plasma membrane docks end to end with a connexon in the membrane of a closely apposed cell. Steps in the synthesis, assembly and turnover of gap junction channels appear to follow the general secretory pathway for membrane proteins. In addition to homo-oligomeric connexons, different connexin polypeptide subunits can also assemble as hetero oligomers. The ability to form homotypic and heterotypic channels that consist of two identical or two different connexons, respectively, adds even greater versatility to the functional modulation of gap junction channels. Electron cryocrystallography of recombinant gap junction channels has recently provided direct evidence for alpha-helical folding of at least two of the transmembrane domains within each connexin subunit. The potential to correlate the structure and biochemistry of gap junction channels with recently identified human diseases involving connexin mutations makes this a particularly exciting area of research. PMID- 9729746 TI - Ion-channel-forming colicins. AB - Several features of ion-channel-forming colicins have been illuminated by recent revelations: its four-domain structure, the mechanism and thermodynamics of binding to the gating loop of outer membrane porins, the mechanism of translocation, competition for the transperiplasmic excursion facilitated by the Tol or Ton transperiplasmic proteins, and the formation of a waisted, funnel shaped transmembrane channel of well-characterized shape. PMID- 9729747 TI - Race/ethnicity differences in the prevalence of noise-induced hearing loss in a group of metal fabricating workers. AB - The National Institute of Occupational Safety and Health rates noise-induced hearing loss as one of the top 10 work-related problems, involving at least 11 million workers. This retrospective study examines the differences between pure tone hearing loss and race/ethnicity in 216 white and 70 non-white male metal fabricating workers. Significant variables upon univariate analysis found to be associated with race/ethnicity were mean years of employment and proportion of time worked without hearing protection. Among whites, the permanent threshold average for 1, 2, 3 and 5 kHz was 25.99 dB, compared with 17.71 dB in non-whites (P < 0.01). Backwards stepwise regression indicated that race/ethnicity, after being adjusted for years of employment, was the major-effect variable. The results of this study suggest that occupational noise exposure alone does not alone account for the racial hearing differences. PMID- 9729748 TI - Occupational noise-induced hearing loss surveillance in Michigan. AB - Occupational noise-induced hearing loss (NIHL) is an important yet often overlooked illness that can affect an individual's safety and performance at work. This article describes a state-based surveillance system for occupational NIHL. The Michigan surveillance system enables us to describe the magnitude of occupational NIHL among Michigan workers and direct public health interventions in the form of enforcement workplace inspections. The data presented are based on interviews of individuals with occupational NIHL reported to the Michigan Department of Consumer and Industry Services (MDCIS) by Michigan's audiologists and otolaryngologists from 1992-1997. From 1992-1997, 1378 individuals with occupational NIHL were reported to the MDCIS and interviewed about their exposures to noise at work. Over 70% of the workplace noise exposure were in manufacturing. At the most recent company where these individuals were exposed to noise, approximately 46% were not provided regular hearing testing. Regular hearing testing was more likely to occur in the larger companies and in industries covered by regulations requiring such testing to be performed. There were improvements over time in the percentages of companies providing regular hearing testing and hearing protection. Construction workers are employees among a group of industries that are not adequately protected from excessive noise exposures by occupational regulations. Regular hearing testing was not provided for over 90% of construction jobs, although hearing protection such as earplugs or earmuffs was provided for approximately half of these jobs. Forty-three state enforcement inspections were conducted at the companies reported by the patients interviewed, because these companies were reported to provide no regular hearing testing or no hearing protection despite exposures to excessive levels of noise. During the 43 inspections, 23 companies had noise levels above dBA, and 17 of those had either no hearing conservation program (HCP) or had one that was cited as being incomplete. The inspections potentially protected 758 similarly exposed workers in the companies with the high noise levels that lacked an HCP or that had a deficient HCP. The number of patients with occupational NIHL is likely a gross underestimate of the true magnitude of the disease. However, the surveillance system has identified workplaces with hazardous levels of noise and no HCP, thereby protecting similarly exposed coworkers of the index patients from further exposures to noise and hearing loss. PMID- 9729749 TI - Cancer incidence among Finnish oil refinery workers, 1971-1994. AB - Cancer incidence between 1971 and 1994 was studied in a cohort of 7,512 men and 1,942 women who had been employed for at least 3 months in a Finnish enterprise that was primarily active in oil refining. The expected numbers of cancer cases were based on the national incidence rates. The standardized incidence ratios (SIR) for overall cancer after 5 years at work was decreased by 12% because of a significant deficit from cancer of the lung in oil refineries (SIR, 0.3; 95% confidence interval [95% CI], 0.1-0.6). There was a significant excess of kidney cancer in males, which was highest among men with at least 5 years of employment in oil refineries (SIR 2.8; 95% CI, 1.6-4.7). Male blue-collar workers had a twofold risk of non-Hodgkin's lymphoma and non-melanocytic skin cancer. Occupational exposure to gasoline may be associated with increased risk of cancer of the kidney. PMID- 9729750 TI - Occupational history collection by third-year medical students during internal medicine and surgery inpatient clerkships. AB - Occupational history is fundamental for the evaluation of possible workplace influences on health. We reviewed 2,922 initial history-and-physical reports from 137 third-year medical students to examine occupational history collection. Overall reporting frequencies were recorded as the following: industry, 55.8%; occupation, 70.0%; specific occupational exposure, 8.4%; smoking status, 91.4%. Patients younger than 40 years of age and women were significantly less likely than other older patients and men to have notations of occupation and industry. Surgery students were less likely than internal medicine students to collect data for industry (41.6% vs 66.6%, P < 0.001), occupation (57.4% vs 79.7%, P < 0.001), and smoking (88.1% vs 94.0%, P < 0.001). The highest frequencies of notation were those for circulatory and respiratory conditions. No significant differences were noted for student gender, academic quarter, or week of clerkship. Clinical occupational medicine teaching should emphasize the need to collect occupational information from all patients, including women and young persons. PMID- 9729751 TI - Health surveillance using an occupational medical database. AB - This pilot study sought associations between liver function tests (LFTs) and membership in homogeneous exposure groups (HEGs) at a target plant as pre clinical indications of possible future occupational health problems. A large company database yielded linear models for each of six LFTs (total bilirubin, alkaline phosphatase, gammaglutamyl transferase, lactate dehydrogenase, aspartate transaminase, and alanine transaminase) in terms of sex, body mass index, age, race (white/non-white), alcohol and cigarette consumption, and production/non production (P/NP) job, permitting control for these in analyses of LFTs vs HEGs at the plant. These analyses, with HEG substituted for P/NP in the large group model, resulted in loosely "suspect" associations significant at P < 0.10. Collapsed HEG variable (containing "suspects" separately and all other non significant HEG levels pooled) yielded "confirmed suspects" at P < 0.05 in the analysis of an independent LFT set taken at the plant approximately one year later. PMID- 9729752 TI - Source, routes, and frequency of pesticide exposure among farmers. AB - Little data on the prevalence of acute and chronic pesticide exposure among farmers and the frequency of protective gear use are available. This report presents results of a cross-sectional study of pesticide behaviors among dairy farmers certified to apply pesticides to field crops. Private dairy farmers residing in six adjacent Wisconsin counties were randomly selected and contacted by telephone for interview. One hundred ninety-one farmers completed the interview (response rate, 76.4%). Twelve percent reported never or almost never wearing gloves during application, and 53.7% reported never or almost never wearing other protective gear. Thirty-two percent reported dermal exposure, and 32.0% reported inhaling pesticides during last application. Findings suggest that exposure to pesticides was frequent in this sample, whereas use of personal protective equipment was not routine. PMID- 9729753 TI - Evaluation of a worksite-based patient education intervention targeted at employees with diabetes mellitus. AB - Diabetes mellitus (DM) affect 5% to 10% of Americans and is estimated to account for $45 billion in direct and $47 billion in indirect costs. The average medical care cost in 1992 for a person with diabetes was $11,157, compared with $2,600 for a person without diabetes. The Diabetes Control and Complications Trial (DCCT) demonstrated that good control of diabetes can delay the onset and slow the progression of many diabetic complications and thereby result in avoidance of costs related to such complications. First Chicago NBD evaluated a worksite-based patient education program for employees with DM. The goal of the program was to determine if such a program could result in improved DM control. A total of 53 employees participated in baseline laboratory testing and met monthly with a diabetic health educator. After 3 months, 45 (85%) employees participated in retesting. One of the 45 employees had an active infection and was not included in the analysis. Of the 44 remaining employees, 43% were on oral agents, 39% on insulin, 2% on combination therapy, and 16% controlled with diet alone. Prior to receiving any laboratory results, 48% of participants rated their level of control as "good" or "very good", while only 9% considered it "poor". After 3 months of educational programs, the subjects' mean fasting blood glucose levels fell from 197.8 mg% to 179.6 mg% (P = 0.12), mean glycohemoglobin declined from 11.5% to 10.1% (P < 0.001), and mean hemoglobin A1C declined from 9.0% to 8.3% (P < 0.001). A worksite-based diabetes disease education program has been shown to significantly improve control of the disease. This should result in lower direct and indirect health care costs and enhanced quality of life. PMID- 9729754 TI - Surveillance of non-fatal workplace assault injuries, using police and employers' reports. AB - Although the majority of work-related homicides are routinely reported in the United States, information on non-fatal events is less complete. Comprehensive surveillance of non-fatal events depends on understanding reporting trends to different agencies. This study characterizes workplace assaults reported to police and through employers in eight southern California cities. Employers' reports filed from October 1, 1994, through January 31, 1995, and police reports filed from June 1, 1994, through March 31, 1995, that involved a non-fatal workplace assault injury were included. Reports from police and employers were linked, and annualized rates combining non-duplicative reports were calculated and event characteristics compared. The combined annualized rate of workplace assault injury for the eight cities was 184.6 per 100,000 workers, which was almost twice the rate found in either reporting source individually. Police reports differed from employers' reports by industry and occupation of victim but not type of event or weapons used. Examination of multiple reporting sources for non-fatal workplace assault injuries is essential to identifying the magnitude of these events. Understanding trends in reporting is important for the effective design of prevention programs. PMID- 9729755 TI - Surveillance of occupational diseases in the United States. A survey of activities and determinants of success. AB - Managers of state-based occupational disease surveillance programs were interviewed for information on their program's characteristics and factors that contributed to their success. There were 68 programs in 52 jurisdictions (50 states, the District of Columbia and New York City). Reportable conditions ranged from a specific disease to "all occupational diseases". Of these programs, 56% met at least one of their objectives. Conditions associated with successful programs usually had short latency periods, were easily diagnosed, and were related to a workplace hazard. They included agricultural injuries, burns, respiratory diseases, cumulative trauma disorders, and poisonings due to lead, pesticides, or carbon monoxide. Successful programs had larger budgets and more staff than did unsuccessful programs, and also took actions after notification of a condition. PMID- 9729756 TI - Occupational injuries among older workers with visual, auditory, and other impairments. A validation study. AB - This study aims to validate a previously defined model of the risk of occupational injuries among older workers with visual, auditory, or other impairments. That model was based upon the Health and Retirement Study (HRS). The previous logistic regression model was recalculated using data from the 1994 National Health Interview Survey (NHIS). The parameter estimates for impaired hearing (.181 in NHIS, 1.55 in HRS), impaired vision (2.42 in NHIS, 1.48 in HRS), and self-employment (0.22 in NHIS, 0.49 in HRS) were in same direction and of roughly the same magnitude. The previously defined model was confirmed using NHIS data. The data suggest that as the workforce ages, more attention must be paid to the accommodation of disabilities in the workplace, especially sensory impairments-poor vision and hearing. PMID- 9729757 TI - Taking dental self-care to the extreme: 24-month incidence of dental self extractions in the Florida Dental Care Study. AB - OBJECTIVE: A common response to health-related symptoms is to treat oneself in lieu of or prior to seeking formal health care. Among the more extreme forms of dental self-care is dental self-extraction. To our knowledge, no study of the incidence of this behavior has been conducted. The objective of this study was to determine if one form of dental self-care, dental self-extraction, is a real phenomenon, and if so, to determine its incidence. METHODS: The Florida Dental Care Study is a longitudinal study of changes in oral health, whose subjects participated for an interview and clinical examination at baseline and 24 months after baseline. RESULTS: Of the 739 persons who participated through 24 months 176 lost one or more teeth. Of these 176 persons, 13 (7%) extracted one or more of their own teeth. The clinical status at baseline of the self-extracted teeth was consistent with the ability to self-extract. CONCLUSION: The phenomenon of dental self-extraction is real and is not limited to residents of developing nations or geographically isolated areas. Because of the potential for prolonged bleeding or bacterial endocarditis in certain population groups, community health clinicians and officials should be cognizant of this behavior. PMID- 9729758 TI - Type 1 diabetes mellitus and oral health: assessment of tooth loss and edentulism. AB - OBJECTIVE: The oral health of an adult population previously diagnosed with juvenile onset insulin dependent-diabetes was comprehensively assessed. The goal of this exploratory cross-sectional evaluation was to described the characteristics related to partial tooth loss edentulism in subjects with Type 1 diabetes mellitus. METHODS: An adult population of 406 Type 1 diabetes mellitus subjects, who had been monitored for 6-8 years as part of a University of Pittsburgh longitudinal study of medical complications associated with diabetes, received an oral health examination for missing teeth, edentulism, coronal and root caries, periodontal status, and oral health behaviors. RESULTS: Of the 406 subjects evaluated, 204 had no missing teeth, 186 had partial tooth loss (1-27 missing teeth), and 16 were edentulous. Patients who had partial tooth loss or who were edentulous were generally older; had lower incomes and levels of education; and had higher rates of nephropathy, neuropathy, retinopathy, and peripheral vascular disease. A logistic regression model found partial tooth loss to be significantly associated with extensive periodontal disease in remaining teeth (OR = 7.35), a duration of diabetes longer than 24 years (OR = 5.32), not using dental floss (OR = 2.37), diabetic neuropathy (OR = 2.29), household income less than $20,000 (OR = 2.21), multiple coronal caries and fillings (OR = 1.98), and bleeding on probing (OR = 1.82). CONCLUSIONS: Although the majority of these adult Type 1 diabetes patients had serious medical complications associated with their diabetes, the possible impact of diabetes mellitus on oral health should be included in their overall management. PMID- 9729763 TI - The John W. Knutson Distinguished Service Award in Dental Public Health--1997 Recipient John C. Greene. PMID- 9729764 TI - The intracellular life of Chlamydia psittaci: how do the bacteria interact with the host cell? AB - Throughout the life of any organism interactions with the surrounding environment are always taking place, a process that leads to evolution. Chlamydia psittaci is an obligate intracellular parasite, but it must also be capable of extracellular survival in order to search for new host cells. Therefore, these peculiar prokaryotes have evolved two different particles and a unique developmental cycle that, together with a series of not yet fully understood interactions with their host cells, allow them to fulfil the requirements for their permanence in nature. These interactions are the subject of this paper. Particular attention is paid to the attachment and internalization of the bacteria, the chlamydial vacuole, and the avoidance of lysosomal degradation. PMID- 9729765 TI - Horizontal gene transfer from transgenic plants to terrestrial bacteria--a rare event? AB - Today, 12 years after the first field release of a genetically modified plant (GMP), over 15,000 field trials at different locations have been performed. As new and unique characteristics are frequently introduced into GMPs, risk assessment has to be performed to assess their ecological impact. The possibilities of horizontal gene transfer (HGT; no parent-to-offspring transfer of genes) from plants to microorganisms are frequently evaluated in such risk assessments of GMPs before release into the field. In this review we indicate why putative HGT from plants to terrestrial (soil and plant associated) bacteria has raised concern in biosafety evaluations. Further, we discuss possible pathways of HGT from plants to bacteria, outline the barriers to HGT in bacteria, describe the strategies used to investigate HGT from plants to bacteria and summarize the results obtained. Only a few cases of HGT from eukaryotes such as plants to bacteria have been reported to date. These cases have been ascertained after comparison of DNA sequences between plants and bacteria. Although experimental approaches in both field and laboratory studies have not been able to confirm the occurrence of such HGT to naturally occurring bacteria, recently two studies have shown transfer of marker genes from plants to bacteria based on homologous recombination. The few examples of HGT indicated by DNA sequence comparisons suggest that the frequencies of evolutionarily successful HGT from plants to bacteria may be extremely low. However, this inference is based on a small number of experimental studies and indications found in the literature. Transfer frequencies should not be confounded with the likelihood of environmental implications, since the frequency of HGT is probably only marginally important compared with the selective force acting on the outcome. Attention should therefore be focused on enhancing the understanding of selection processes in natural environments. Only an accurate understanding of these selective events will allow the prediction of possible consequences of novel genes following their introduction into open environments. PMID- 9729766 TI - Tricarboxylic acid cycle and anaplerotic enzymes in rhizobia. AB - Rhizobia are a diverse group of Gram-negative bacteria comprised of the genera Rhizobium, Bradyrhizobium, Mesorhizobium, Sinorhizobium and Azorhizobium. A unifying characteristic of the rhizobia is their capacity to reduce (fix) atmospheric nitrogen in symbiotic association with a compatible plant host. Symbiotic nitrogen fixation requires a substantial input of energy from the rhizobial symbiont. This review focuses on recent studies of rhizobial carbon metabolism which have demonstrated the importance of a functional tricarboxylic acid (TCA) cycle in allowing rhizobia to efficiently colonize the plant host and/or develop an effective nitrogen fixing symbiosis. Several anaplerotic pathways have also been shown to maintain TCA cycle activity under specific conditions. Biochemical and physiological characterization of carbon metabolic mutants, along with the analysis of cloned genes and their corresponding gene products, have greatly advanced our understanding of the function of enzymes such as citrate synthase, oxoglutarate dehydrogenase, pyruvate carboxylase and malic enzymes. However, much remains to be learned about the control and function of these and other key metabolic enzymes in rhizobia. PMID- 9729767 TI - Comparative mapping of the two wheat leaf rust resistance loci Lr1 and Lr10 in rice and barley. AB - The wheat genome is large, hexaploid, and contains a high amount of repetitive sequences. In order to isolate agronomically important genes from wheat by map based cloning, a simpler model of the genome must be used for identifying candidate genes. The objective of this study was to comparatively map the genomic regions of two wheat leaf rust disease resistance loci, Lr1 and Lr10, in the putative model genomes of rice and barley. Two probes cosegregating with the Lr1 gene on chromosome 5DL of wheat were studied. The rice sequences corresponding to the two probes were isolated and mapped. The two probes mapped to two different rice chromosomes, indicating that the organization of the region orthologous to Lr1 is different in rice and wheat. In contrast, synteny was conserved between wheat and barley in this chromosomal region. The Lrk10 gene cosegregated with Lr10 on chromosome 1AS in wheat. The rice gene corresponding to Lrk10 was mapped on rice chromosome 1, where it occurred in many copies. This region on rice chromosome 1 corresponds to the distal part of the group 3S chromosomes in Triticeae. The synteny is conserved between rice chromosome 1 and the Triticeae group 3S chromosomes up to the telomere of the chromosomes. On group 3S chromosomes, we found a gene that is partially homologous to Lrk10. We conclude that in the genomic regions studied, there is limited and only partially useful synteny between wheat and rice. Therefore, barley should also be considered as a model genome for isolating the Lr1 and Lr10 genes from wheat. PMID- 9729768 TI - A study of variation in rDNA ITS regions shows that two haplotypes coexist within a single aphid genome. AB - We report variation in the rDNA internal transcribed spacers (ITSs) of aphid species, the first for these insects. Variation at 6 sites within ITS1 sequences of the green peach aphid, Myzus persicae, identified two haplotypes coexisting within the same individuals, indicating that molecular drive has not homogenised different copies of rDNA. During this study, we found that PCR can cause a precise 58-bp loss in the amplified copies of an ITS haplotype (type 1). This occurs in all detectable copies under routine PCR conditions, at different annealing temperatures and with Pfu and Taq polymerases. In addition, "hot-start" PCR exclusively copied a different, rare haplotype (type 2). These observations have important considerations for using PCR, as large deletions in PCR products may not reflect real deletions in the genome, and changes in PCR conditions may be needed to copy cryptic haplotypes. PMID- 9729770 TI - Characterisation and physical localisation of Ty1-copia-like retrotransposons in four Alstroemeria species. AB - The genus Alstroemeria contains species with large genomes (2C = 36.5-78.9 pg (17,600-38,000 Mb) in those species with 2n = 2x = 16). We investigated the diversity and genomic and chromosomal organisation of Ty1-copia-like retrotransposons in four Alstroemeria species. Analysis of 33 PCR-amplified sequences corresponding to a conserved domain of the Ty1-copia reverse transcriptase (rt) gene showed high heterogeneity among predicted amino acid sequences; no two sequences were identical, but most fell into one of five subgroups. Levels of inter- and intra-specific heterogeneity of sequences were similar. HaeIII-digested genomic DNA of various Alstroemeria species contained distinct bands upon hybridisation with individual rt gene fragments. Hybridisation with the heterogeneous PCR pool of rt fragments (retrotransposon pool) revealed additional bands; some minor bands were characteristic of either Brazilian or Chilean species. In situ hybridisation of the retrotransposon pool from three species to metaphase chromosomes from the same species showed a dispersed distribution of the retrotransposon pool with exclusion from rDNA and other chromosomal sites. Alstroemeria pelegrina, which is without major heterochromatic sites, showed some clustering and small negative bands. The retrotransposon pool was excluded from major DAPI-staining bands in Alstroemeria aurea, but in contrast, the sites of the major tandemly repeated sequences in Alstroemeria inodora showed a hybridisation signal similar to that in the rest of the chromosomes. The data are discussed in the context of the contribution of Ty1 copia-like retrotransposons to plant genome size, their evolution, and their value for phylogenetic and biodiversity studies. PMID- 9729772 TI - A genetic and molecular analysis of an invectedDominant mutation in Drosophila melanogaster. AB - The invected gene of Drosophila melanogaster is a homeobox-containing gene that is closely related to engrailed. A dominant gain of function allele, invectedDominant, was derived from mutagenesis of a dominant allele of vestigial, In(2R)vgW. A careful analysis of the phenotype of invectedDominant shows that it is associated with a transformation of the anterior compartment of the wing to a posterior fate. This transformation is normally limited to the wing blade itself and does not involve the remaining tissues derived from the wing imaginal disc, including the wing hinge and dorsal thorax of the fly. The ectopic expression of invected protein associated with invectedDominant correlates spatially with the normal expression pattern of vestigial in the wing imaginal disc, suggesting that control elements of vestigial are driving ectopic invected expression. This was confirmed by sequence analysis that shows that the dominant vestigial activity was eliminated by a deletion that removes the 3' portion of the vestigial coding region. This leaves a gene fusion wherein the vestigial enhancer elements are still juxtaposed immediately 5' to the invected transcriptional start site, but with the vg sequences harboring an additional lesion. Unlike recessive invected alleles, the invectedDominant allele produces an observable phenotype, and as such should prove useful in determining the role of invected in patterning the wing imaginal disc. Genetic analysis has shown that mutations of polyhomeotic, a gene involved in regulating engrailed expression, cause a reproducible alteration in the invectedDominant phenotype. Finally, the invectedDominant allele should prove valuable for identifying and characterizing genes that are activated within the posterior compartment. A screen using various lacZ lines that are asymmetrically expressed in an anterior-posterior manner in the wing imaginal disc isolated one line that shows posterior-specific expression within the transformed anterior compartment. PMID- 9729774 TI - Drosophila melanogaster histone H2B retropseudogene is inserted into a region rich in transposable elements. AB - We have isolated and characterized the genomic sequence of a Drosophila melanogaster histone H2B pseudogene that is localized outside of the cluster of the replication-dependent histone genes and has all the properties of a retropseudogene. It is highly homologous to the transcribed region of the D. melanogaster histone H2B gene, but not to its flanking regions, and is surrounded by short direct repeats. The pseudogene contains several point mutations that preclude its translation. The sequence of the 3' region of this pseudogene is compatible with the hypothesis that the 3' terminal stem-loop structure of the histone H2B mRNA has served as a primer for the reverse transcription event from which this pseudogene originated. Analysis of the regions flanking the histone H2B pseudogene revealed the presence of three different types of transposable elements, suggesting that this chromosomal locus represents a hotspot for transposition. PMID- 9729777 TI - Self-amplification of satellite DNA in vitro. AB - We describe a PCR-like reaction in which genomic DNA acts as a template as well as a primer. Interaction between genomic tandem repeat units leads to self amplification of satellite DNA. This genomic self-priming PCR (GSP-PCR) allowed the rapid amplification of species-specific tandem repeats of horse, cattle, dolphin, and chicken. A novel specific satellite of ostrich with a repeat unit of 60 bp was isolated using this method. PMID- 9729781 TI - Characterization of microsatellite loci from Elymus alaskanus and length polymorphism in several Elymus species (Triticeae: Poaceae). AB - A size-selected genomic library from Elymus alaskanus was screened for the presence of (GA)n, (GT)n, (CAC)n, and (TCT)n microsatellite sequences. A total of 28 positive clones were found for the two dinucleotide repeats, whereas no positive clones were found for the trinucleotide repeats. Positive clones were sequenced to validate the presence of microsatellites and to generate polymerase chain reaction (PCR) primers, based on the sequences flanking the microsatellite. Primer pairs were designed and evaluated for 18 selected microsatellites. The resulting loci were analysed by PCR for their usefulness as molecular markers in an array of 18 accessions representing E. alaskanus and 10 other Elymus species. PCR amplification revealed alleles for the 18 loci, which varied in having 1-10 alleles in these accessions. In the 18 accessions tested, 7 of the 18 loci were polymorphic, with gene diversity values ranging from 0.54 to 0.80 among all species. Within E. alaskanus, gene diversity values ranged from 0.20 to 0.72, with a mean of 0.48. Polymorphism was also detected within accessions. No clear relationship between total repeat length and the degree of polymorphism was observed in this study. Primer pairs designed to amplify microsatellites in E. alaskanus can be used to generate polymorphism products in other species within the genus. Hence, microsatellites are useful markers for studying both inter- and intra-specific genetic variability within Elymus. PMID- 9729783 TI - FISH mapping and inter-Alu fingerprinting define the YAC contig map around the centromeric region of human chromosome 18. AB - Previous reports concerning the location of D18S44 with respect to the centromere have been ambiguous. Also, it has not been possible, based on formerly reported markers, to show that contigs WC18.0 and WC18.1 overlap. However, the data presented here definitively show, using FISH technology, that D18S44 (located on WC18.0) maps to proximal 18q. Furthermore, inter-Alu fingerprinting shows a clear overlap between WC18.0 and WC18.1, thereby establishing a complete contig between D18S44 and markers from WC18.1. PMID- 9729784 TI - Characterization and structure of the mitochondrial small rRNA gene of the entomopathogenic fungus Beauveria bassiana. AB - The entire mitochondrial (mt) small ribosomal RNA (srRNA) gene from the entomopathogenic fungus Beauveria bassiana was sequenced. Alignment of the sequence to those of other filamentous fungi revealed gross length differences in their respective products. Construction of a secondary structural model showed that these differences were restricted to known variable srRNA subdomains. Several features were identified that were common only to the hyphomycetous fungi examined. Phylogenetic analysis indicated that the anamorph B. bassiana was more closely related to the pyrenomycete than to the plectomycete ascomycetous fungi. Based on our previous comparison of mt gene arrangement in filamentous fungi, this was unexpected. The possibility that the smaller mt genomes reflect the ancestral arrangement of genes is discussed. PMID- 9729785 TI - Recommendations for the presentation of NMR structures of proteins and nucleic acids. IUPAC-IUBMB-IUPAB Inter-Union Task Group on the Standardization of Data Bases of Protein and Nucleic Acid Structures Determined by NMR Spectroscopy. AB - The recommendations presented here are designed to support easier communication of NMR data and NMR structures of proteins and nucleic acids through unified nomenclature and reporting standards. Much of this document pertains to the reporting of data in journal articles; however, in the interest of the future development of structural biology, it is desirable that the bulk of the reported information be stored in computer-accessible form and be freely accessible to the scientific community in standardized formats for data exchange. These recommendations stem from an IUPAC-IUBMB-IUPAB inter-union venture with the direct involvement of ICSU and CODATA. The Task Group has reviewed previous formal recommendations and has extended them in the light of more recent developments in the field of biomolecular NMR spectroscopy. Drafts of the recommendations presented here have been examined critically by more than 50 specialists in the field and have gone through two rounds of extensive modification to incorporate suggestions and criticisms. PMID- 9729786 TI - NMR with 13C, 15N-doubly-labeled DNA: the Antennapedia homeodomain complex with a 14-mer DNA duplex. AB - Nearly complete 1H, 13C and 15N NMR assignments have been obtained for a doubly labeled 14-base pair DNA duplex in solution both in the free state and complexed with the uniformly 15N-labeled Antennapedia homeodomain. The DNA was either fully 13C, 15N-labeled or contained uniformly 13C, 15N-labeled nucleotides only at those positions which form the protein-DNA interface in the previously determined NMR solution structure of the Antennapedia homeodomain-DNA complex. The resonance assignments were obtained in three steps: (i) identification of the deoxyribose spin systems via scalar couplings using 2D and 3D HCCH-COSY and soft-relayed HCCH COSY; (ii) sequential assignment of the nucleotides via 1H-1H NOEs observed in 3D 13C-resolved NOESY; and (iii) assignment of the imino and amino groups via 1H-1H NOEs and 15N-1H correlation spectroscopy. The assignment of the duplex in the 17 kDa protein-DNA complex was greatly facilitated by the fact that 1H signals of the protein were filtered out in 13C-resolved spectroscopy and by the excellent carbon chemical shift dispersion of the DNA duplex. Comparison of corresponding 13C chemical shifts of the free and the protein-bound DNA indicates conformational changes in the DNA upon complex formation. PMID- 9729787 TI - Experiments and strategies for the assignment of fully 13C/15N-labelled polypeptides by solid state NMR. AB - High-resolution heteronuclear NMR correlation experiments and strategies are proposed for the assignment of fully 13C/15N-labelled polypeptides in the solid state. By the combination of intra-residue and inter-residue 13C-15N correlation experiments with 13C-13C spin-diffusion studies, it becomes feasible to partially assign backbone and side-chain resonance in solid proteins. The performance of sequences using 15N instead of 13C detection is evaluated regarding sensitivity and resolution for a labelled dipeptide (L-Val-L-Phe). The techniques are used for a partial assignment of the 15N and 13C resonances in human ubiquitin. PMID- 9729789 TI - Internal and overall motions of the translation factor eIF4E: cap binding and insertion in a CHAPS detergent micelle. AB - The mRNA cap-binding protein eIF4E is the limiting factor in the eIF4F translation initiation complex, which mediates the binding of the 40S ribosome to the mRNA. 15N relaxation studies have been used to characterize the backbone dynamics of deuterated eIF4E in a CHAPS micelle for the apoprotein, the m7GDP bound form, and the dinucleotide (m7GpppA)-bound form, as well as for CHAPS-free eIF4E. Large differences in overall correlation time between the CHAPS-free form (11.8 ns) and samples containing different concentrations of CHAPS (15.9-19.4 ns) indicate that eIF4E is embedded in a large micelle in the presence of CHAPS, with a total molecular weight in the range of 40-60 kDa. CHAPS seems to restrict the mobility of the a2-b3 and a4-b5 loops which are thought to be embedded in the micelle. No significant changes in overall mobility were seen between the m7 GDP bound form, the m7GpppA-bound form, and the apoprotein. Amide hydrogen exchange data indicate the presence of slowly exchanging amides in two surface-exposed helices (a2 and a4), as well as the a4-b5 loop, indicating protection by the CHAPS micelle. The micelle covers the convex side of the protein away from the cap-binding site. PMID- 9729788 TI - Chemical shift as a probe of molecular interfaces: NMR studies of DNA binding by the three amino-terminal zinc finger domains from transcription factor IIIA. AB - We report the NMR resonance assignments for a macromolecular protein/DNA complex containing the three amino-terminal zinc fingers (92 amino acid residues) of Xenopus laevis TFIIIA (termed zf1-3) bound to the physiological DNA target (15 base pairs), and for the free DNA. Comparisons are made of the chemical shifts of protein backbone 1HN, 15N, 13C alpha and 13C beta and DNA base and sugar protons of the free and bound species. Chemical shift changes are analyzed in the context of the structures of the zf1-3/DNA complex to assess the utility of chemical shift change as a probe of molecular interfaces. Chemical shift perturbations that occur upon binding in the zf1-3/DNA complex do not correspond directly to the structural interface, but rather arise from a number of direct and indirect structural and dynamic effects. PMID- 9729790 TI - Complete assignment of 1H, 13C and 15N chemical shifts for bovine beta lactoglobulin: secondary structure and topology of the native state is retained in a partially unfolded form. AB - Although beta-lactoglobulin (beta-LG) has been studied extensively for more than 50 years, its physical properties in solution are not yet understood fully in terms of its three-dimensional (3D) structure. For example, despite a recent high resolution crystal structure, it is still not clear why the two common variants of bovine beta-LG which differ by just two residues have different aggregation properties during milk processing. We have conducted solution-state NMR studies on a recombinant form of the A variant of beta-LG at low pH conditions where the protein is partially unfolded and exists as a monomer rather than a dimer. Using a 13C, 15N-labelled sample, expressed in Pichia pastoris, we have employed the standard combination of 3D heteronuclear NMR techniques to obtain near complete assignments of proton, carbon and nitrogen resonances. Using a novel pulse sequence we were able to obtain additional assignments, in particular those of methyl groups in residues preceding proline within the sequence. From chemical shifts and on the basis of inter-residue NOEs, we have inferred the secondary structure and topology of monomeric beta-LG A. It includes eight antiparallel beta-strands arranged in a barrel, flanked by an alpha-helix, which is typical of a member of the lipocalin family. A detailed comparison with the crystal structure of the dimeric form (for a mixture of A and B variants) at pH 6.5 reveals a close resemblance in both secondary structure and overall topology. Both forms have a ninth beta-strand which, at the higher pH, forms part of the dimer interface. These studies represent the first full NMR assignment of beta-LG and will form the basis for a complete characterisation of the solution structure and dynamics of this protein and its variants. PMID- 9729791 TI - A new general method for the biosynthesis of stable isotope-enriched peptides using a decahistidine-tagged ubiquitin fusion system: an application to the production of mastoparan-X uniformly enriched with 15N and 15N/13C. AB - A new strategy is described for the production of peptides enriched with stable isotopes. Peptides of interest are expressed in Escherichia coli (E. coli) cells as recombinant fusion proteins with Saccharomyces cerevisiae ubiquitin. This method yields as much as 30-100 mg/l of isotope-enriched fusion proteins in minimal media. A decahistidine tag attached to the N-terminus of ubiquitin enables a one-step purification of the fusion protein via Ni(2+)-chelating affinity chromatography. The ubiquitin moiety is then easily and specifically cleaved off by a protease, yeast ubiquitin hydrolase. Since this enzyme is also expressed at a high level in E. coli cells and can be purified in one step, the presented strategy has an advantage in view of costs over others that use commercially available proteases. In addition, since ubiquitin fusion proteins easily refold, the fusion protein can be expressed either in a soluble form or as inclusion bodies. This flexibility enables us to prepare peptides that are unstable in a soluble state in E. coli cells. As an example, the expression and the uniform stable isotope enrichment with 15N and/or 13C are described for mastoparan-X, a tetradecapeptide known to activate GTP-binding regulatory proteins. An amide group at the C-terminus of this peptide can also be formed by our method. The presented system is considered powerful for the stable isotope enrichment of short peptides with proton resonances that are too severely overlapped to be analyzed solely by proton NMR. PMID- 9729792 TI - Conformational analysis of a Chlamydia-specific disaccharide alpha-Kdo-(2-->8) alpha-Kdo-(2-->O)-allyl in aqueous solution and bound to a monoclonal antibody: observation of intermolecular transfer NOEs. AB - The disaccharide alpha-Kdo-(2-->8)-alpha-Kdo (Kdo: 3-deoxy-D-manno-oct-2-ulosonic acid) represents a genus-specific epitope of the lipopolysaccharide of the obligate intracellular human pathogen Chlamydia. The conformation of the synthetically derived disaccharide alpha-Kdo-(2-->8)-alpha-Kdo-(2-->O)-allyl was studied in aqueous solution, and complexed to a monoclonal antibody S25-2. Various NMR experiments based on the detection of NOEs (or transfer NOEs) and ROEs (or transfer ROEs) were performed. A major problem was the extensive overlap of almost all 1H NMR signals of alpha-Kdo-(2-->8)-alpha-Kdo-(2-->O)-allyl. To overcome this difficulty, HMQC-NOESY and HMQC-trNOESY experiments were employed. Spin diffusion effects were identified using trROESY experiments, QUIET-trNOESY experiments and MINSY experiments. It was found that protein protons contribute to the observed spin diffusion effects. At 800 MHz, intermolecular trNOEs were observed between ligand protons and aromatic protons in the antibody binding site. From NMR experiments and Metropolis Monte Carlo simulations, it was concluded that alpha-Kdo-(2-->8)-alpha-Kdo-(2-->O)-allyl in aqueous solution exists as a complex conformational mixture. Upon binding to the monoclonal antibody S25-2, only a limited range of conformations is available to alpha-Kdo (2-->8)-alpha-Kdo-(2-->O)-allyl. These possible bound conformations were derived from a distance geometry analysis using transfer NOEs as experimental constraints. It is clear that a conformation is selected which lies within a part of the conformational space that is highly populated in solution. This conformational space also includes the conformation found in the crystal structure. Our results provide a basis for modeling studies of the antibody disaccharide complex. PMID- 9729793 TI - Comparison of 15N- and 13C-determined parameters of mobility in melittin. AB - Backbone and tryptophan side-chain mobilities in the 26-residue, cytolytic peptide melittin (MLT) were investigated by 15N and 13C NMR. Specifically, inverse-detected 15N T1 and steady-state NOE measurements were made at 30 and 51 MHz on MLT at 22 degrees C enriched with 15N at six amide positions and in the Trp19 side chain. Both the disordered MLT monomer (1.2 mM peptide at pH 3.6 in neat water) and alpha-helical MLT tetramer (4.0 mM peptide at pH 5.2 in 150 mM phosphate buffer) were examined. The relaxation data were analyzed in terms of the Lipari and Szabo model-free formalism with three parameters: tau m, the correlation time for the overall rotation; S2, a site-specific order parameter which is a measure of the amplitude of the internal motion; and tau e, a local, effective correlation time of the internal motion. A comparison was made of motional parameters from the 15N measurements and from 13C measurements on MLT, the latter having been made here and previously [Kemple et al. (1997) Biochemistry, 36, 1678-1688]. tau m and tau e values were consistent from data on the two nuclei. In the MLT monomer, S2 values for the backbone N-H and C alpha-H vectors in the same residue were similar in value but in the tetramer the N-H order parameters were about 0.2 units larger than the C alpha-H order parameters. The Trp side-chain N-H and C-H order parameters, and tau e values were generally similar in both the monomer and tetramer. Implications of these results regarding the dynamics of MLT are examined. PMID- 9729794 TI - NMR detection of slow conformational dynamics in an endonuclease toxin. AB - The cytotoxic activity of the secreted bacterial toxin colicin E9 is due to a non specific DNase housed in the C-terminus of the protein. Double-resonance and triple-resonance NMR studies of the 134-amino acid 15N- and 13C/15N-labelled DNase domain are presented. Extensive conformational heterogeneity was evident from the presence of far more resonances than expected based on the amino acid sequence of the DNase, and from the appearance of chemical exchange cross-peaks in TOCSY and NOESY spectra. EXSY spectra were recorded to confirm that slow chemical exchange was occurring. Unambiguous sequence-specific resonance assignments are presented for one region of the protein, Pro65-Asn72, which exists in two slowly exchanging conformers based on the identification of chemical exchange cross-peaks in 3D 1H-1H-15N EXSY-HSQC, NOESY-HSQC and TOCSY HSQC spectra, together with C alpha and C beta chemical shifts measured in triple resonance spectra and sequential NH NOEs. The rates of conformational exchange for backbone amide resonances in this stretch of amino acids, and for the indole NH of either Trp22 or Trp58, were determined from the intensity variation of the appropriate diagonal and chemical exchange cross-peaks recorded in 3D 1H-1H-15N NOESY-HSQC spectra. The data fitted a model in which this region of the DNase has two conformers, NA and NB, which interchange at 15 degrees C with a forward rate constant of 1.61 +/- 0.5 s-1 and a backward rate constant of 1.05 +/- 0.5 s-1. Demonstration of this conformational equilibrium has led to a reappraisal of a previously proposed kinetic scheme describing the interaction of E9 DNase with immunity proteins [Wallis et al. (1995) Biochemistry, 34, 13743-13750 and 13751 13759]. The revised scheme is consistent with the specific inhibitor protein for the E9 DNase, Im9, associating with both the NA and NB conformers of the DNase and with binding only to the NB conformer detected because the rate of dissociation of the complex of Im9 and the NA conformer, NAI. is extremely rapid. In this model stoichiometric amounts of Im9 convert, the E9 DNase is converted wholly into the NBI form. The possibility that cis-trans isomerisation of peptide bonds preceding proline residues is the cause of the conformational heterogeneity is discussed. E9 DNase contains 10 prolines, with two bracketing the stretch of amino acids that have allowed the NA [symbol: see text] NB interconversion to be identified, Pro65 and Pro73. The model assumes that one or both of these can exist in either the cis or trans form with strong Im9 binding possible to only one form. PMID- 9729795 TI - Sequence-specific 1H assignment and secondary structure of the bacteriocin AS-48 cyclic peptide. AB - The bacteriocin AS-48 is a cationic peptide (7149 Da) having a broad antimicrobial spectrum, encoded by the 68 kb conjugative plasmid pMB2 from Enterococcus faecalis S-48. It is a unique peptide since it has a cyclic structure, which is achieved by the formation of a tail-head peptide bond after ribosomal synthesis (Galvez et al., 1989; Martinez-Bueno et al., 1994; Samyn et al., 1994). Preliminary CD and calorimetric studies (data not shown) pointed towards a highly helical and very stable three dimensional structure. All the information gathered until now indicates that the target of AS-48 is the cytoplasmic membrane in which it opens channels or pores, leading to dissipation of the proton motive force and cell death, which in some cases is also followed by bacterial lysis (Galvez et al., 1991). This peptide is a suitable tool for studying protein-membrane interactions, and it also offers promising perspectives for biotechnological applications. Knowledge of the 3D structure of AS-48 is a first step in the conduct of further structure-function studies. Here we report the complete 1H NMR assignment of its proton resonances together with the resulting secondary structure pattern as prerequisites for the determination of a high-resolution 3D solution structure. PMID- 9729796 TI - Local helix content and RNA-binding activity of the N-terminal leucine-repeat region of hepatitis delta antigen. AB - Hepatitis delta virus (HDV) is a satellite virus of the hepatitis B virus (HBV) which provides the surface antigen for the viral coat. Our results show that the N-terminal leucine-repeat region of hepatitis delta antigen (HDAg), encompassing residues 24-50, binds to the autolytic domain of HDV genomic RNA and attenuates its autolytic activity. The solution conformation of a synthetic peptide corresponding to residues 24-50 of HDAg as determined by two-dimensional 1H NMR and circular dichroism techniques is found to be an alpha-helix. The local helix content of this peptide was analyzed by NOEs and coupling constants. Mutagenesis studies indicate that Lys38, Lys39, and Lys40 within this alpha-helical peptide may be directly involved in RNA binding. A structural knowledge of the N-terminal leucine-repeat region of HDAg thus provides a molecular basis for understanding its role in the interaction with RNA. PMID- 9729797 TI - Sequential assignment of the triple labelled 30.1 kDa cell-adhesion domain of intimin from enteropathogenic E. coli. PMID- 9729798 TI - Assignment of 1H, 13C, and 15N signals of turkey ovomucoid third domain at pH 2.0. PMID- 9729799 TI - 1H, 15N and 13C resonance assignments and secondary structure of apo liver fatty acid-binding protein. PMID- 9729800 TI - 1H, 13C and 15N NMR backbone assignments of 25.5 kDa metallo-beta-lactamase from Bacteroides fragilis. PMID- 9729801 TI - A convenient method for affinity purification of maltose binding protein fusions. AB - Maltose binding protein of Escherichia coli is frequently employed as a fusion partner for the biosynthesis of polypeptides and proteins, largely because of the advantage provided by affinity purification of such a fusion. A mixture of cellulose and starch is shown to be a simple and effective alternative to the use of other, more complicated media which are often used for affinity chromatography of maltose binding protein fusions. PMID- 9729802 TI - Effect of the replication mode of a plasmid on the stability of multimeric endoxylanase genes in Bacillus subtilis. AB - Effect of the replication mode of a plasmid on the stability of tandemly multimerized endoxylanase genes and a gene dose-dependent expression of the endoxylanase were studied in Bacillus subtilis. The structural genes encoding an endoxylanase, carrying its original promoter and ribosomal binding sequence, were tandemly multimerized and cloned into the Escherichia coli-B. subtilis shuttle plasmid, pJH27 delta 88 or pMTL500e, which has a rolling circle-replicon or a theta (theta)-replicon in B. subtilis, respectively. The cloned dimers in pJH27 delta 88, which has a rolling circle-replicon, spontaneously rearranged to monomers in B. subtilis DB104, whereas those in pMTL500e, having a theta (theta) replicon, were stably maintained. Expression level of the endoxylanase was proportional to the gene dosage in multimers. The endoxylanase activity in the supernatant increased from 80 U ml-1 with pMTL-1x containing a monomer of the gene to 165 U ml-1 with pMTL-4x containing a tetramer. These results indicate that high level expression of the endoxylanase gene can be obtained by tandemly multimerizing the genes in a plasmid with a theta (theta)-replicon. PMID- 9729803 TI - A comparison of intensive cell culture bioreactors operating with hybridomas modified for inhibited apoptotic response. AB - It is demonstrated, using two different perfusion reaction systems, that hybridoma modified by inhibiting their apoptotic response can give improved process performance in terms of cell number and viability in intensive cell culture. Two cell perfusion systems, one using a spin filter and the other an ultrasonic filter, are compared using two cell lines. One cell line is transfected with the bcl-2 gene (TB/C3 bcl-2) which encodes the 'anti-apoptotic' human bcl-2 protein and the other cell line (TB/C3 pEF) with a negative transfection vector. Both reactor systems give similar retention performance for both cell lines. Bcl-2 transfected cells reach higher cell densities than the control cell line, and the percentage of apoptotic cells is clearly lower than with pEF cells. The maximum cell numbers of the bcl-2 cell line are 1.21 x 10(7) ml-1 in the ultrasonic filter culture and 1.58 x 10(7) ml-1 in the spin filter culture, respectively. Using the pEF cell line the maximum cell number reaches 6.0 x 10(6) ml-1 with ultrasonic retention and 5.9 x 10(6) ml-1 in the spin filter. The use of ultrasound in this cell retention system has no apparent influence on cell growth, productivity or viability. Selective retention of viable cells is detectable but the effect of removing non-viable cells is negligible. PMID- 9729804 TI - Bio-dissolution of spent nickel-cadmium batteries using Thiobacillus ferrooxidans. AB - In this study, the production of sulphuric acid in bioreactors with Thiobacillus ferrooxidans attached on elemental sulphur was investigated. These bioreactors reached a maximum H+ productivity of 80 mmol kg-1 d-1 of support. This medium was used for the indirect dissolution of spent nickel-cadmium batteries recovering after 93 days 100% of cadmium, 96.5% of nickel and 95.0% of iron. Moreover, recoveries higher than 90.0% were reached when anodic and cathodic materials were directly added to Thiobacillus ferrooxidans cultures with sulphur as the sole energy source. The results presented show an economic and effective method which could be considered the first step to recycle spent and and discarded batteries preventing one of the many problems of environmental pollution. PMID- 9729805 TI - Copper-dependent depolymerization of lignin in the presence of fungal metabolite, pyridine. AB - Thus far, it has not been recognized that copper complexes are able to depolymerize lignin under physiological conditions of white rot decay. However, we have found that both phenolic and non-phenolic synthetic lignins were intensively depolymerized by Cu(II) and lipid hydroperoxide model compounds in the presence of a metabolite of ligninolytic fungi, pyridine at room temperature in aqueous media. Treatment of 14C-labeled oxygen-prebleached kraft pulp (OKP) by the copper-dependent reaction evidenced effectiveness of this reaction for the delignification of kraft pulps. In contrast to the organic peroxide system, Cu(II)/pyr/H2O2 system was much less effective for the lignin depolymerization. However, treatment of unbleached kraft pulp (UKP) by Cu(II)/H2O2 and Cu(II)/pyr/H2O2 systems demonstrated that the damage of cellulose was suppressed by the coordination of pyridine although high brightness gain was obtained independently of the presence of the coordinator. Spin trapping experiments demonstrated that not hydroxyl radical but superoxide anion is involved in the Cu(II)/pyr/H2O2 system. This finding not only introduces a new concept of non enzymatic lignin biodegradation by wood-degrading fungi but also presents a new strategy for decomposing lignin and lignin-related compounds by copper complexes and peroxide-producing system. PMID- 9729807 TI - Comparison of macroscopic cranial methods of age estimation applied to skeletons from the Terry Collection. AB - A total of 963 skeletons (408 Whites and 555 Blacks) from the Terry Collection were studied to examine macroscopic cranial methods of age estimation. The methods of Acsadi and Nemeskeri, Masset, Baker and Meindl and Lovejoy were applied to every skull. The results indicate that the most accurate techniques in this application were those that consider endocranial suture closure. The methods of Acsadi and Nemeskeri and Masset were the most accurate in all the subsamples (by population, sex, sex within population and in total), although the relative accuracy could vary in application to other populations. PMID- 9729806 TI - The spectroscopic analysis, inhibition and binding studies demonstrate the equivalence of Erythrina caffra trypsin inhibitor and the recombinant substitution variant recSerETI. AB - A recombinant substitution mutant (recSerETI) of the Erythrina caffra trypsin inhibitor, with the N-terminal valine residue substituted by serine, was produced in E. coli and compared to the wildtype protein (wtETI) with respect to physicochemical and functional properties. The spectral properties, including UV absorbance, fluorescence emission and circular dichroism, were indistinguishable. Furthermore, the inhibitory activities of the two proteins regarding the inhibition of trypsin, chymotrypsin, tissue plasmininogen activator (t-PA) and reteplase (BM 06.022, t-PA deletion variant comprising the kringle 2 and the protease domains, isolated from transformed E. coli cells) and the affinity of the immobilized inhibitors for reteplase were closely similar. Five repetitive cycles of guanidinium chloride (GdmCl)-induced denaturation-renaturation yield the native mutant protein with its inhibitory activity fully restored. The only difference between the wildtype and the mutant protein refers to the intrinsic stability. Comparing the pH- and GdmCl-dependent transitions, as well as the thermal denaturation, recSerETI exhibits decreased stability compared to the wildtype protein. The pH range of stability is shifted from pH 1-9.5, for wtETI, to pH 2-9, for recSerETI; similarly the GdmCl-induced denaturation is found to occur at a GdmCl half concentration of 3.7 M instead of 4.5 M; in both cases the renaturation exhibits strong hysteresis. The mid-point of the thermal unfolding transition of the mutant protein is at approximately 65 degrees C, as compared to approximately 75 degrees C for the wildtype protein. PMID- 9729808 TI - Biological and chemical hazards of forensic skeletal analysis. AB - In the course of conducting forensic analysis of human skeletal material, anthropologists are exposed to a number of biological and chemical hazards. This paper reviews the primary concerns in terms of infection or exposure. Handling of human tissue provides an avenue through which bloodborne pathogens may be transported. Scene recovery also includes a set of hazards through exposure to human, animal and soil vectors. Basic personnel protection and laboratory procedures should be established for the protection of all personnel involved in this work. PMID- 9729809 TI - Evaluation of aspartic acid racemization ratios in the human femur for age estimation. AB - Levels of D-aspartic acid (D/L ratio) in cranial non-collagen proteins (acid soluble peptide fractions) have been reported to increase with age. We isolated total amino acid fractions from the femur and separately isolated acid-insoluble collagen fraction and acid-soluble peptide fractions; then D/L ratios were measured from each fraction by gas chromatography. We evaluated the applicability of their D/L ratios for age estimation based on their correlation coefficient. A sex-related difference was observed in the D/L ratio. In particular, aged females showed a low ratio, suggesting an association with bone disorders. In males, the D/L ratios of acid-soluble peptide fraction showed the highest correlation rate (r = 0.969) with age, and those of total amino acid fraction showed the highest correlation rate (r = 0.633) with age in females. Without separation of male and female, the D/L ratios of total amino acid fraction showed the highest value (r = 0.853). The D/L ratio of acid-soluble peptide fractions differed according to the size of bone powder particles, being higher for larger particle sizes. These results suggest that the application of D/L ratio from total amino acid fraction is the most effective method for estimating age using the human femur. However, care is necessary when studing cadavers that might be females. PMID- 9729810 TI - Metric and comparative analysis of sexual dimorphism in the Thai femur. AB - Identification of sex from the skeleton is an important demographic assessment in medicolegal investigations. Studies have demonstrated that populations differ from each other in size and proportions and that these differences can affect the metric assessment of sex. It is therefore vital to determine if population differences are great enough to necessitate group-specific standards. To date, there have been no attempts to create standards of assessment for modern Thais. Therefore the purpose of this research is to establish standards from which to determine sex from the femur using a new skeletal collection housed at the Chiang Mai University Department of Anatomy. The sample is composed of 104 individuals (70 males, 34 females). Six standard osteometric dimensions including maximum length, maximum head diameter, midshaft circumference, midshaft anterior posterior and transverse diameters, and bicondylar breadth were measured and analyzed by stepwise discriminant function statistics. To understand population differences, formulas derived from Chinese, South African whites and American whites and blacks using the same method and variables were tested on the Thai sample. Results indicated that maximum head diameter and bicondylar breadth are the optimal combination for sex diagnosis and yielded 94.2% accuracy. Direct analysis using predetermined single or multiple variables also revealed bicondylar breadth as the best single dimension (93.3%). In cross-tests on the Thais, the Chinese formula gave the most favorable outcome with unsatisfactory results for all other groups. The present research confirms that sexual dimorphism is better reflected in breadth dimensions than in bone length. Comparisons showed that Thais are very different metrically from whites and blacks, and although they most resemble the Chinese, these two groups are not identical. These findings underscore the need for population-specific formulas for identification of sex from the skeleton. PMID- 9729811 TI - Identification of race and sex from palate dimensions. AB - Measurements of the width and depth of the palate were used to predict the race (American black or white) or sex or both of an individual. The sample consisted of 332 living subjects with permanent dentitions, and measurements were made between cusp tips, so palate size includes bony and dental components. Blacks, with a more square palate, were distinguished from whites primarily by greater interpremolar widths and P1-to-M2 depths. Simultaneous prediction of race and sex had a correct classification of 48%, which is about twice that expected from chance. Pooling the two sex increases correct classification of race to 83%. Formulas also are provided for each variable separately to accommodate fragmentary remains. Resilience of palatal structures to traumatic and natural forces makes this method practical in several forensic situations. PMID- 9729812 TI - Suicide among youth and young adults, 15 through 24 years of age. A report of 392 cases from Paris, 1989-1996. AB - The aims of our study were (1) to examine the socio-demographic, clinical characteristics, autopsy and toxicological findings in 392 youth suicides in Paris, between 1989 through 1996, and (2) to analyze the psychodynamic determinants leading up to the onset of the suicide. During the eight-year study period 392 suicides involving young people were investigated at the Institute of Forensic Medicine of Paris. Two hundred and sixty victims (66%) were males. The mean age was 22 years in both sexes. Fifteen percent of the victims were below 20 years. Ninety-two percent of the subjects were single. Forty percent of the victims were students, 35% were unemployed. One third of the victims had previously attempted suicide. Thirty-five percent of the subjects used to take psychoactive prescription drugs and some of them had been under the care of a mental health professional at the time of the suicide. In 40% of the cases a suicide note was found near the body. Depression (70% of victims), schizophrenia, (10%), affective disorders, parent-child relational problems, partner relational problems, adolescent antisocial behavior, and borderline personality were found to be the most frequent diseases and stressors involved in the suicides. The suicide was rarely an accidental reaction to stress. It was constantly preceded by situational distress, which led to suicidal ideas if the adolescent failed to cope with problems. Ten percent were known as heroin users. In more than 40% of the cases, the victim's parents were divorced or separated. The most frequent method of suicide was poisoning followed by jumping from a height, gunshot, subway death, and hanging/asphyxia. Among firearms, a handgun was more likely to be used than rifles (85/15%). Tranquilizers were the most frequent psychoactive drugs used for suicide followed by antipsychotic drugs, antidepressants, and barbiturates (10%). PMID- 9729813 TI - Implantable cardioverter-defibrillators and the pathologist: comment and cautionary notes. AB - This paper briefly reviews the components of, the clinical uses of, the techniques to place, and the complications related to implantable cardioverter defibrillators (ICDs). Information useful in the specific identification of ICDs is presented. A series of recommendations for the autopsy examination or postmortem explantation of ICDs by the pathologist is given. Because of the serious risk of injury to the pathologist possible with postmortem discharges of ICDs which have not been deactivated, and because of the risk of device explosion if the ICD is incinerated, a number of cautionary notes are provided. A brief case with occurrence of accidental postmortem discharge of an active ICD is also presented. PMID- 9729814 TI - Identification of cocaine analytes in fingernail and toenail specimens. AB - Fingernail and toenail specimens were obtained from 18 suspected cocaine users. The nails were cut, heated under methanolic reflux, and the methanolic extracts were purified by solid-phase extraction. Gas chromatography/mass spectrometry was utilized for the qualitative and quantitative analysis of nine cocaine analytes. Comparison of conventional postmortem analysis of blood and urine with nail analysis revealed a marked increase in the detection of cocaine use by nail analysis. Cocaine analytes were present in 14 (82.3%) subjects utilizing nail analysis. Out of those 14 subjects, only 5 (27.7%) were positive by conventional postmortem drug analysis. Cocaine and benzoylecgonine were the predominant analytes in all positive nail specimens. Anhydroecgonine methyl ester, ecgonine methyl ester, ecgonine ethyl ester, cocaethylene, norcocaine, and norbenzoylecgonine were detected in a limited number of specimens. The ratio of cocaine to benzoylecgonine ranged from 2-10:1. These findings suggest that nails may be a useful alternative matrix for the detection of cocaine exposure. PMID- 9729815 TI - Medicolegal implications of drugs and chemicals detected in intracranial hematomas. AB - The purpose of this study was to determine how drug findings in intracranial hematomas should be assessed in forensic autopsy cases. Six cases in which intracranial hematomas containing drugs and chemicals were detected were examined in this study. Of the six cases, five were positive for drugs and chemicals that had been self-administered by the victims prior to injury. Post-traumatic time interval from injury to death was in the range 10 to 65 h. In two individuals who were positive for norephedrine or toluene, the concentrations of these substances were much higher in the intracranial hematomas than in heart blood. In an individual who was positive for phenobarbital, its concentration was only a little higher in the intracranial hematoma than in heart blood. In the remaining two cases, substantial quantities of ethanol were detected in the intracranial hematomas, but little ethanol was detected in heart blood. In three cases, some drugs were administered at hospital after the injuries. The time interval from the initial drug administration to death was 19 to 60 h. In two individuals given phenytoin and/or lidocaine intravenously, substantial amounts of these drugs were detected in the intracranial hematomas. In an individual given diazepam intravenously, a substantial quantity of diazepam was detected in heart blood, but not in the intracranial hematoma. Toxicological analysis of intracranial hematomas may be useful not only for determining whether individuals were under the influence of ethanol at the time they were injured, but also for detecting pre-traumatic usage of other drugs and chemicals. However, the medical record should be reviewed thoroughly from a toxicological view point if victims underwent medical treatment prior to death because drugs administered for the purpose of medical treatment can disseminate into preexisting intracranial hematomas, depending on the size of the hematomas. PMID- 9729816 TI - Forensic implications and medical-legal dilemmas of maternal versus fetal rights. AB - The purpose of this paper is to review the issue of fetal rights from primarily a legal perspective, with consideration of morals and professional ethics. The practice of medicine is fraught with numerous bioethical dilemmas. These dilemmas often leave the physician wondering if he has made the correct decision. A physician's morals and professional ethics may influence his or her decision in resolving bioethical dilemmas. The case example is a 34-year-old female with a 41 week intra-uterine pregnancy. The mother was refusing induction of labor. Without the labor induction, the fetus may die. Despite this risk, the mother desired to pursue a vaginal delivery. The AMA's ethics state that a competent, pregnant mother's wishes should prevail and the court should not be involved unless there are unusual circumstances. The mother in the case example was competent and informed consent was provided. Case law does not specifically address the dilemma of the case example. However, there is case law regarding court-ordered cesarean sections which reveals different opinions. The difference in court opinion encompasses the relative degree of weight given to the fetus's right to be born healthy and alive versus the mother's privacy rights. Some courts describe this "balancing test," whereas others state that the mother's privacy rights prevail unless there are exceptional circumstances, which will be extremely rare. The fetus has acquired rights in other areas of the law; for example, abolishment of the intra-family immunity doctrine and the definition of murder in most states. In considering the legal arena of fetal versus maternal rights, a decision tree is presented to assist physicians in assessing cases of a pregnant mother refusing medical treatment. There is no precise demarcation in assessing fetal and maternal rights. The greater the degree of fetal viability, the greater degree of fetal rights. Consideration must also be given to the relative degree of invasiveness to the mother for the proposed procedure; the more invasive, the greater degree of maternal rights. Each case must be evaluated on an individual basis and the decision tree can assist a clinician with this process. PMID- 9729817 TI - Differences in criminal activity between heroin abusers and subjects without psychiatric disorders--analysis of 578 detainees in Bilbao, Spain. AB - The association between drug abuse and criminal activity has been deeply established, but the nature of this relationship is controversial. The incidence and types of criminal activity were analyzed in 837 arrests of 578 subjects who were also interviewed for psychiatric diagnosis and evaluation of criminal responsibility. There was a significant prevalence of heroin abuse/dependence (50.5%) in the sample. Another 124 subjects (21.5%) in whom no psychiatric disorder could be observed were considered as the control group. Heroin abusers were younger (26 years, SD 5.9) than controls (29 years, SD 11.2) and showed some different ethnic characteristics. Heroin abuse/dependence was the most important risk factor (O.R. = 10.15) for criminal recidivism. Females were more related to nonviolent criminal activity than males. There was a higher incidence of offenses against property among heroin abusers (burglary 57.8%; robbery 19.5%) than in the control group (burglary 15.3% robbery 4.8%). In contrast, aggression or resistance to police authorities and nonfatal offenses against persons were more frequent among controls (12% and 13.7%, respectively) than among heroin abusers (3.7% and 3%, respectively). The results of this study confirm the hypothesis of a relationship between criminal activity and heroin abuse/dependence, probably based on financial needs. However, the association seems not to be a single and direct cause-effect relationship, as other factors show influence on the criminal activity. PMID- 9729818 TI - Efficacy of repeated psychophysiological detection of deception testing. AB - Physiological measures were recorded during repeated psychophysiological detection of deception (PDD) tests to determine if reaction levels change with test repetition. Two groups of 22 healthy male subjects completed six peak of tension PDD tests on each of two test days. A minimum between test day interval of six days was maintained. The treatment group was programmed to respond deceptively to one of seven test questions while the control group was programmed to respond truthfully to all questions. The respiration and galvanic skin resistance (GSR) line lengths, GSR peak response amplitude and latency, and cardiovascular inter-beat-interval (IBI) were calculated for each response. Analyses indicated that, except for GSR peak response latency, differential physiological reactivity during a PDD test did not change significantly over repeated tests or days; there was a decrease in average respiration line lengths at the initial test(s) of each day; and differential changes in average respiration line length, GSR peak latency, and cardiovascular IBI responses corresponded to deception. Power analyses were calculated to assist in result interpretation. It is suggested that PDD decision accuracy, concerning subject veracity, should not decrease during repeated testing. PMID- 9729819 TI - QIAamp spin columns as a method of DNA isolation for forensic casework. AB - The Detroit Police Crime Lab has historically used Chelex as a method to isolate DNA for amplification and typing of bloodstains at the HLADQA1, PM and D1S80 loci. However, preliminary validation of several STR systems for casework has demonstrated that the Chelex procedure is not the best method of DNA isolation for STR amplifications for our purposes. Long term storage at -20 degrees C in the presence of unbuffered Chelex beads (approximately 1 year), combined with multiple freeze thaws, resulted in signal loss at a locus for many database samples. Therefore, we have employed the QIAamp spin column as an alternative method of DNA isolation for amplification and typing of STR loci currently being validated for use in the laboratory. Moreover, we determined that QIAamp isolated DNA is also suitable for HLADQA1, PM and D1S80 typing. A matrix study was performed to determine if the QIAamp DNA procedure would give better results on bloodstains deposited on "problem surfaces" such as leather, dirt and various dyed fabrics. Again, QIAamp isolated DNA was more readily typeable than Chelex isolated DNA. We successfully replaced the phenol/chloroform extraction steps utilized in our laboratory for differential extractions, a commonly used method for separating sperm and non-sperm fractions of sexual assault evidence, with the QIAamp spin columns. The QIAamp extracted DNA performed as well in all PCR amplification and typing procedures tested (PM, HLADQA1, D1S80, and STR (PowerPlex)) as the phenol/chloroform Centricon isolated or EtOH precipitated DNAs. Thus we concluded that QIAamp spin columns are a superior method for isolating DNA to be typed for a variety of loci. PMID- 9729820 TI - South Portuguese population data on the loci HLA-DQA1, LDLR, GYPA, HBGG, D7S8 and Gc. AB - Five South Portuguese Caucasian subpopulations were analyzed for the HLA-DQA1, LDLR, GYPA, HBGG, D7S8 and Gc loci. Genotype distributions for these loci did not deviate from Hardy-Weinberg expectations. The allele and genotype frequencies found have been compared with previously published data from North and Central Portugal. A total of 11 out of 138 chi-square comparisons of allele frequencies between different Portuguese populations showed a certain degree of divergence. Alentejo, Algarve, Madeira Island and Azores Islands populations might be considered as different groups in a database. For forensic casework, a composite South Portuguese Caucasian population database was obtained for estimating multiple locus profile frequencies using the six PCR-based loci studied. PMID- 9729821 TI - New Jersey Caucasian, African American, and Hispanic population data on the PCR based loci HLA-DQA1, LDLR, GYPA, HBGG, D7S8, and Gc. AB - New Jersey Caucasian, African American, and Hispanic genotype and allele frequencies were determined for the six PCR-based loci, HLA-DQA1, LDLR, GYPA, HBGG, D7S8, and Gc. All but one locus (HLA-DQA1 for African Americans) meet Hardy Weinberg expectations. However, observing one departure in 18 loci over the three New Jersey sample populations is not unexpected. There is little evidence for departures from independence between pairs of loci in the three populations studied. Thus, multiple locus profile frequencies can be determined using the product rule. PMID- 9729822 TI - The effect of pathologic substances and adulterants on the DNA typing of urine. AB - Human urine has not been adequately investigated as a potential source of DNA for forensic identity testing. The advent of polymerase chain reaction technology has made possible the analysis of previously undetectable levels of nucleic acids from human urine and other body fluids lacking nucleated cells. In this study, we evaluated the ability to genotype DNA extracted from adulterated urine specimens using the AmpliType PM + DQA1 PCR amplification and typing system. Fresh, first void male urine specimens were contaminated with household bleach, E. coli, human serum albumin, glucose and saponin (a strong detergent). All of the adulterated samples were typed without difficulty. Frozen male urine specimens were split into equal volumes; one aliquot was adulterated with either E. coli or saponin, and the other was left free of contaminants. Seventy-one percent of all frozen urine specimens tested (adulterated and unadulterated) were successfully typed using this amplification and typing system. Our data, therefore, suggest that the AmpliType PM + DQA1 PCR amplification and typing system described is suitable for genotype analysis of adulterated fresh and frozen urine specimens. PMID- 9729823 TI - Development of a highly polymorphic STR marker for identity testing purposes at the human androgen receptor gene (HUMARA). AB - We developed a non-isotopic method which improves the technical quality of the X linked HUMARA locus typing process. The use of formamide and a low concentration of acrylamide increased resolution and sharpness of HUMARA alleles in silver stained polyacrylamide gels. In addition, the construction of an allelic ladder containing amplified sequence of 9 alleles (even-numbered alleles) of the HUMARA locus, allows confident, rapid and precise assignment of discretely defined alleles. Allele and genotype frequencies for the HUMARA locus were determined in a French Canadian population sample. Observed genotype frequencies in females conformed to Hardy-Weinberg expectations. Furthermore, the HUMARA locus is highly polymorphic with 18 observed alleles and an heterozygosity value of 89.3%. Also, this locus has average powers of discrimination of 97.8% and 88.7% for testing samples of female and male origin, respectively. In the French Canadian population, the average probability of excluding a random man as the father in paternity analysis when both mother and daughter are tested for this locus is 88.0%. Together, the results indicate that the HUMARA locus provides a highly discriminatory system that is appropriate for the purposes of forensic identification and paternity testing involving a female child. PMID- 9729824 TI - Computer-based production of bite mark comparison overlays. AB - Bite mark comparison protocols include measurement and analysis of the pattern, size, and shape of teeth against similar characteristics observed in an injury on skin or a mark on an object. The physical comparison of tooth position often depends upon transparent acetate overlays to detect similarities or differences between the teeth and the bite mark. Several methods are used to produce life sized comparison overlays. The perimeter of the biting edges of the anterior teeth are usually recorded to produce facsimile images called hollow volume overlays. Some investigators hand-trace these outlines from dental study casts, or from bite exemplars produced in wax, styrofoam, or similar materials. Some use hand-traced outlines from xerographic images produced with office photocopiers that are calibrated to produce life-sized final images. Others use radiographic images and toneline photography of wax exemplars filled with radio-opaque materials, such as metal filings or barium sulfate. Dependence upon subjective input by the odontologist to trace these images manually is considered problematic. This is because the errors incorporated at any production stage are increased in the final product. The authors have developed a method to generate accurate hollow volume overlays using computer-based techniques. A PowerPC Macintosh computer, flatbed scanner, and Adobe Photoshop (a popular graphical interface application) are used to acquire, select, arrange and export detailed data from class and individual characteristics of a suspect's teeth to acetate film loaded in a high-resolution laser printer. This paper describes this technique to enable the odontologist to produce high-quality, accurate comparison overlays without subjective input. PMID- 9729826 TI - Determination of benzodiazepines in forensic samples by HPLC with photo-diode array detection. AB - A high-performance liquid chromatographic (HPLC) method has been developed for the analysis of several benzodiazepines and some of their metabolites in blood, plasma and urine. The method included a liquid-liquid extraction with n hexane:ethylacetate, a gradient elution on a C8 reversed phase column with a non electrolyte eluent and a photo diode array detection. This allowed a rapid detection, a purity check, and identification as well as quantitation of the eluting peaks. The detection limit was 10 to 30 ng and the limit of quantitation was 0.05 microgram/mL, using 1 mL of blood, plasma or urine. The procedure is applied routinely in forensic toxicological analyses involving blood, stomach content, urine and organ samples. About 30 positive cases are reported. The avoidance of the use of an electrolyte buffer in the eluent resulted in a robust procedure, free of technical problems and of long rinsing periods, suitable for routine use in forensic toxicology analysis involving blood, urine, stomach content and tissue samples. PMID- 9729825 TI - Immersion technique for brain removal in perinatal autopsies. AB - Perinatal autopsies present forensic patholgists with a variety of challenges, not the least of which involves the removal and examination of very small and sometimes fragile organs. Removal of the immature brain can be particularly troublesome. Even if great care is taken during brain removal, one is often left with no more than a semifluid amorphous mass of softened tissue by the time the brain is ready to be fixed in formalin. We describe a method of perinatal brain removal which helps to preserve brain shape and integrity. By removing the brain while the head (and body) is totally immersed in water, we find that the brain is easier to remove and less apt to destruction. Subsequent fixation in formalin results in well-preserved, intact specimens, allowing for optimal examination and sectioning. PMID- 9729827 TI - Postmortem diagnosis of leukodystrophies. AB - Leukodystrophies are progressive disorders involving the development and maintenance of myelin in the central and peripheral nervous systems. Although relatively uncommon, leukodystrophic disorders may be undiagnosed or misdiagnosed during life, and may appear as "sudden death." In such instances, these victims may be referred to a forensic pathologist. In general, leukodystrophies are inherited in an autosomal recessive manner so that proper postmortem diagnosis by the forensic pathologist is extremely important to the decendant's family for future family planning. PMID- 9729828 TI - Sudden death due to sarcoid heart disease. AB - A case of sudden death due to massive myocardial sarcoidosis is presented. Cardiac sarcoidosis is discussed. Since the deceased was a New York City police officer with death benefit entitlements under the Heart Bill, the implications of the medicolegal autopsy are emphasized. PMID- 9729829 TI - Speedballing with needle embolization: case study and review of the literature. AB - Foreign-body embolization is not an uncommon occurrence. However, to our knowledge, there are only ten reported cases of needle embolization associated with intravenous drug use. We report the sudden death of a 49-year-old white male with a known history of crack cocaine abuse. At autopsy, suspicious needle marks were noted on the right lower extremity. The lungs were of increased weight at 1000 and 1090 g and appeared edematous. The heart weighed 520 g and had a normal red-brown myocardium. Upon sectioning, a broken hypodermic needle of very small caliber was identified in the right ventricular myocardium protruding into the right ventricular chamber. This needle apparently traveled from the injection site to the right ventricle. The right ventricle was dilated and hypertrophied, and microscopic examination showed hyperemic myocardium surrounding the needle. Sections of lung showed numerous foreign-body type giant cells containing polarizable foreign material consistent with intravenous drug use. Toxicological analysis revealed the presence of ethanol (36 mg/dL), cocaine (0.098 mg/L), benzoylecgonine (2.16 mg/L), and morphine (0.841 mg/L). Urine and blood were positive for the presence of 6-monoacetylmorphine. Based on the toxicological analysis, the cause of death was determined to be cocaine and heroin toxicity, and the manner accidental. The needle embolus was considered an incidental finding. PMID- 9729830 TI - Airbag mediated death of a two-year-old child wearing a shoulder/lap belt. AB - Airbag injuries have resulted in the deaths of several infants and small children, and such deaths are generally associated with rearward-facing infant seats or unrestrained children in front passenger seats of cars equipped with airbags. An airbag can also cause death in a small child wearing a shoulder/lap belt, however, as this case report illustrates. A two-year-old female was involved in a low-speed collision while riding in the front passenger seat of a dual-airbag-equipped automobile. Secondary impact with the airbag caused catastrophic occipitoatlantoaxial disarticulation with traumatic spinal cord separation, thermal injury and abrasions of the right forearm and distinctive patterned abrasions of the face. The possibility of airbag injury should be considered in all low-speed traffic fatalities, and the confirmatory injuries sought at postmortem examination. PMID- 9729831 TI - A fatal drug interaction between clozapine and fluoxetine. AB - A case is presented of a fatal drug interaction caused by ingestion of clozapine (Clozaril) and fluoxetine (Prozac). Clozapine is a tricyclic dibenzodiazepine derivative used as an "atypical antipsychotic" in the treatment of severe paranoid schizophrenia. Fluoxetine is a selective serotonin reuptake inhibitor used for the treatment of major depression. Clinical studies have proven that concomitant administration of fluoxetine and clozapine produces increased plasma concentrations of clozapine and enhances clozapine's pharmacological effects due to suspected inhibition of clozapine metabolism by fluoxetine. Blood, gastric, and urine specimens were analyzed for fluoxetine by gas chromatography/mass spectrometry (GC/MS) and for clozapine by gas-liquid chromatography (GLC). Clozapine concentrations were: plasma, 4.9 micrograms/mL; gastric contents, 265 mg; and urine, 51.5 micrograms/mL. Fluoxetine concentrations were: blood, 0.7 microgram/mL; gastric contents, 3.7 mg; and urine 1.6 micrograms/mL. Norfluoxetine concentrations were: blood, 0.6 microgram/mL, and none detected in the gastric contents or urine. Analysis of the biological specimens for other drugs revealed the presence of ethanol (blood, 35 mg/dL; vitreous, 56 mg/dL; and urine 153 mg/dL) and caffeine (present in all specimens). The combination of these drugs produced lethal concentrations of clozapine and high therapeutic to toxic concentrations of fluoxetine. The deceased had pulmonary edema, visceral vascular congestion, paralytic ileus, gastroenteritis and eosinophilia. These conditions are associated with clozapine toxicity. The combined central nervous system, respiratory and cardiovascular depression of these drugs was sufficient to cause death. The death was determined to be a clozapine overdose due to a fatal drug interaction. PMID- 9729832 TI - Distribution of phenol in a fatal poisoning case determined by gas chromatography/mass spectrometry. AB - A victim who was presumed to have ingested waste fluid containing phenol of DNA extraction was found dead in his laboratory. The skin was partially chemically burned, with blisters as maps. No mechanical injuries were observed. The pathological findings of the liver and kidney were typical of those of acute substantial poisoning. Phenol concentrations in the blood, urine, stomach contents and organs were determined by gas chromatography/mass spectrometry. Phenol was distributed throughout the body. The concentration of free phenol in the blood was found to be 60 micrograms/mL, and in the urine it was 208 micrograms/mL. The phenol concentrations in the organs were found as follows: 106 micrograms/g in the brain; 116 micrograms/g in the lungs; 166 micrograms/g in the liver, and 874 micrograms/g in the kidney, respectively. Significantly high concentrations were observed in the kidney, urine, and liver. To the best of our knowledge, such an intoxication through this kind of ingestion has never been reported before. Distributions of phenol in fatal poisonings have been reported, but colorimetry was used as the analytical method and it cannot exclude the interference of other phenolic compounds. PMID- 9729833 TI - Personal identification on the basis of antemortem and postmortem radiographs. AB - This report documents three recent cases in Hungary in which personal identification was achieved by comparison of antemortem and postmortem radiographs. These cases demonstrate three examples of radiological identification. In Hungary, comparison methods play an important role in personal identification because of the lack of adequate dental records for most of the population. The authors emphasize that in cases where antemortem radiographs and photographs are available, radiographic comparison is deemed preferable to photographic superimposition, because it is more technically exacting and permits the matching of a potentially larger number of anatomical, pathological or traumatic features. PMID- 9729834 TI - PM and D1S80 loci gene frequencies in the Zaragoza population of northern Spain. AB - LDLR, GYPA, HBGG, D7S8, GC (PM loci) and D1S80 are widely used in forensic casework analyses and population data are required to estimate the frequency of a DNA profile. This paper presents the results of a survey aimed at investigating the allele and genotype frequency distribution of these loci in an important Spanish population (Zaragoza, North Spain). Statistical analysis to determine whether allele frequencies were in Hardy-Weinberg equilibrium was carried out as well as to obtain some parameters of medicolegal interest. There was no evidence of association between the alleles of the loci. The Zaragoza sample does not differ substantially from other Caucasian populations. PMID- 9729835 TI - Isolation, amplification, and sequencing of human mitochondrial DNA obtained from human crab louse, Pthirus pubis (L.), blood meals. AB - The ability to identify individual human hosts based on analyses of blood recovered from the digestive tract of hematophagous arthropods has been a long term pursuit in both medical and forensic entomology. Blood meal individualization techniques can bring important advancements to studies of vector-borne disease epidemiology. Forensically, these analyses may aid in assailant identification in violent crime cases where blood-feeding insects or their excreta are recovered from victims or at crime scenes. Successful isolation, amplification, and sequencing of human mitochondrial DNA obtained from adult human crab lice fed on human volunteers are reported. Adult lice were removed from recruited volunteers frequenting inner city health clinics. Live lice were killed by freezing and subsequently air dried at ambient temperature. A saliva sample was obtained from each volunteer and served as a DNA reference sample. Volunteers were afforded free, approved pediculosis treatment. Individual lice were subsequently processed using procedures developed for the extraction of mitochondrial DNA from human hair, teeth, and bone. The resulting DNA was amplified by the polymerase chain reaction and sequenced. Our results point to valuable avenues for future entomological research. PMID- 9729837 TI - Adult age differences in semantic and episodic memory. AB - In 2 experiments, performance was compared on memory tests taken by older and younger adults for information that had been rated as more memorable for older adults or more memorable for each age group by adults in that age group. In Experiment 1, semantic knowledge was tested by having the participants recognize or recall information from memory (e.g., name as many states as you can). The older adults performed significantly better; thus, hypotheses of superior knowledge and ease of retrieval of that knowledge for that age group were supported. In Experiment 2, lists of presidents and states were presented to the participants, followed by recognition and recall tests of episodic memory. The younger adults performed significantly better. The older adults studied familiar items on the lists for less time and made many more errors. Thus, the hypothesis of an encoding deficit influenced by item familiarity for older adults was supported. PMID- 9729836 TI - Five-year-olds' behaviorally presented self-esteem: relations to self-perceptions and stability across a three-year period. AB - In this study (a) the connection between 5-year-olds' behaviorally presented self esteem and their domain-specific self-perceptions and (b) the stability in behaviorally presented self-esteem across a 3-year period (between age 5 and age 8) were examined. The behavioral manifestations of self-esteem were rated by the children's teachers using the Behavioral Rating Scale of Presented Self-Esteem in Young Children (J. Haltiwanger & S. Harter, 1988). The children's domain-specific self-perceptions were assessed by the Pictorial Scale for Perceived Competence and Social Acceptance (S. Harter & R. Pike, 1984). Ninety-five 5-year-olds (50 boys and 45 girls) and their teachers participated. Follow-up data on behavioral self-esteem were available for 55 of the children (30 boys and 25 girls). The results showed, first, that 5-year-olds' presented self-esteem as rated by their teachers was positively and significantly related to the children's perceptions of competence, but not to their perceptions of social acceptance. Second, a high degree of stability in presented self-esteem was found between age 5 and age 8. Some implications of these results for educational practice are discussed. PMID- 9729838 TI - Nonresidential father involvement: a test of a mid-range theory. AB - This study tested a theoretical model of postdivorce involvement of nonresidential fathers with their children. The hypotheses were (a) that postdivorce father involvement is related to father parenting role identity, role clarity, child relationship quality, and father role hierarchy ranking; and (b) that several variables serve as moderators for the relationship between father parenting role identity and father involvement. Data were collected via self report questionnaires administered to 101 fathers. Path analytic techniques were used to assess the theoretical model. Findings supported the hypotheses related to the core constructs within the theory, but not for the moderator variables within the study. Role clarity and child relationship quality were found to have both direct and indirect effects on father involvement. Father parenting role identity had a direct effect on father involvement but also functioned as a meditating variable for constructs within the model. Significant paths were found for two variables, joint custody and satisfaction with the legal system. Father postdivorce involvement is a complex phenomenon, and forces external to fathers can contribute to levels of postdivorce involvement (e.g., legal system issues). PMID- 9729839 TI - Effects of perceived attractiveness and academic success on early adolescent peer popularity. AB - Effects of perceived attractiveness and academic performance on 9th graders' ratings of peers' popularity were investigated. Participants were 270 9th graders (152 girls, 118 boys) who read a vignette describing a hypothetical same-sex peer with whom the student had been assigned to complete a project. The partner's attractiveness and academic performance were systematically varied in four conditions: high attractiveness/high grades, high attractiveness/low grades, low attractiveness/high grades, and low attractiveness/low grades. After reading the vignette, the students rated the partner's popularity. As hypothesized, analyses of variance revealed that attractive partners were significantly more popular than unattractive partners, regardless of whether the partner had high or low grades. Contrary to expectation, attractiveness was not more important to girls than to boys. Integration with past research and suggestions for future research are offered. PMID- 9729840 TI - A comparison of two measures to assess adult attachment. AB - The purpose of this study was to determine the relationship between two different approaches to the assessment of adult attachment. The approaches were C. Hazan and P. R. Shaver's (1987) categorization of three attachment styles and the dimensional system of the Reciprocal Attachment Questionnaire (RAQ; M. West & A. Sheldon-Keller, 1992). Participants were 196 (72 men and 124 women) respondents to a community survey. Multiple regression results showed that the three attachment groups differed significantly on three dimensions of the RAQ: feared loss of the attachment figure, perceived availability of the attachment figure, and use of the attachment figure. In addition, men and women differed significantly on use of the attachment figure, independent of their attachment style. The results indicate that the two approaches to the assessment of adult attachment are interrelated in meaningful ways. PMID- 9729841 TI - The relation of problem behaviors in preschool children to depressive symptoms in mothers and fathers. AB - The relation of maternal and paternal depressive symptoms to problem behaviors in a nonclinical sample of preschool children was examined. Data were collected from 46 women, their husbands, and their 4-year-old, first-born children. Observed maternal restrictive and punishing behavior and attachment security of the child were considered additional sources of risk for externalizing and internalizing problem behaviors. Different predictors for child externalizing and internalizing behaviors were identified via hierarchical multiple regression analyses. Maternal and paternal depressive symptoms and maternal restrictive and punishing behavior emerged as salient predictors of child internalizing behaviors. For externalizing behaviors, there were significant gender differences: For girls, maternal depressive symptoms made a significant contribution to the model; the model for boys was not significant. The results perhaps reflect different etiological pathways for externalizing and internalizing behaviors, supporting the suggestion that those behaviors are distinct clinical phenomena, even among very young children. The findings also suggest that nonclinical levels of parental symptomatology show systematic relations to children's problem behaviors. PMID- 9729842 TI - A proposed relationship between circumcision and neural reorganization. AB - Humans are intensely biocultural beings. The linkages and causal feedback loops among their symbolic world, their cultural world, and their physical bodies can be exquisitely complex and subtle. It is suggested in this article that one cultural event--circumcision--exemplifies that subtlety and complexity. It is hypothesized that circumcision reorganizes the male's sensory somato-cortex to raise the threshold of sexual excitability/distraction. This threshold shift thereby allows the young men of a social group (a) to be slightly more tractable in executing corporate activities beneficial to the community and (b) to be slightly more restrained sexually and more cooperative in the pair bond. The practice is accepted because the procedure is deeply enmeshed in the ritual and symbolic life of the social group and is applicable to all young males. Suggestions are made on how to test this hypothesis empirically. PMID- 9729843 TI - Assessing intimacy with the best friend and the sexual partner during adolescence: the PAIR-M inventory (Personal Assessment of Intimacy in Relationship). AB - The present study had 3 goals: (a) to provide a preliminary investigation of the dimensions involved in the capacity for intimacy toward the best friend and the sexual partner during adolescence; (b) to determine whether the specific areas of the capacity for intimacy toward the best friend are the same as toward the sexual partner; and (c) to consider the usefulness of conceiving the capacity for intimacy as a multidimensional concept. Canadian high school students (N = 465; 257 girls, 208 boys) completed a questionnaire on the capacity for intimacy, best friend version; 232 of them completed the partner version of the questionnaire. Factorial analysis on the best friend version of the questionnaire identified 3 factors: Social Intimacy, Positive Intimacy, and Negative Intimacy. Factorial analysis on the partner version of the questionnaire identified 4 factors: Social Intimacy, Positive Intimacy, Negative Intimacy, and Sexual Intimacy. PMID- 9729844 TI - Relationship of maternal and general self-acceptance to pre- and postpartum affective experience. AB - Forty-nine married primiparous Israeli women responded to W. W. K. Zung's (1965) Self-Rating Depression Scale, N. M. Bradburn's (1969) Affect Balance Scale, and measures of general and maternal self-acceptance during the last trimester of pregnancy and again 6 to 8 weeks following childbirth. There was a significant decrease in depression from pre- to postpartum for the total group. Women high in general self-acceptance were less depressed and displayed less negative affect than those low in general self-acceptance. There were no corresponding differences between the high and low maternal self-acceptance groups. Both pre- and postpartum women tended to rate themselves significantly higher for maternal self-acceptance than for general self-acceptance. PMID- 9729845 TI - Premenstrual symptoms in Mexican women with different educational levels. AB - Most women present some premenstrual symptoms, which may be influenced by diverse sociocultural factors. The authors studied the premenstrual symptoms of 89 healthy Mexican women living in rural areas and whose education ranged from no schooling to middle school attendance, and 182 women living in urban areas whose education ranged from elementary school to professional studies. The Menstrual Distress Questionnaire (R. H. Moos, 1968) was completed by all the women during the premenstrual and postmenstrual phases of one menstrual cycle. The percentage of women who reported mild symptoms was 87% on somatic scales and 86% on psychological scales. Premenstrual symptoms were more severe among women engaged in professional studies. Urban women reported more severe psychological symptoms than rural women. When women who were engaged in professional studies were excluded from the data analyses, no differences between the groups were found. Thus, it appears that the women's level of education affected premenstrual symptoms more than their rural or urban backgrounds did. PMID- 9729846 TI - Adolescents' perceptions of paternal and maternal parenting styles in a Chinese context. AB - Chinese secondary school students (N = 429) were asked to respond to instruments measuring their perception of parents' global parenting styles and specific parenting practices. Results showed that there were significant differences between reported paternal parenting and maternal parenting characteristics, with fathers perceived as relatively less responsive, less demanding, less concerned, and more harsh. Adolescent girls' perceptions of fathers' parenting characteristics generally did not differ from those of the boys, but the girls tended to perceive their mothers as more demanding but less harsh. The present findings provide some support for the popular Chinese saying, "strict father, kind mother," but they also suggest that it requires redefinition. PMID- 9729847 TI - The influence of physical state and color on perceived sweetness. AB - Smell, texture, temperature, and other variables can influence the evaluation of foods and beverages. The purpose of this study was to investigate the influence of physical state and color on perceived sweetness. Fifty junior high school students were given 10 samples of an aqueous sucrose solution in liquid and solid (gelatin) form in random order and were asked to rate their sweetness on a 10 point scale. For each state (liquid and solid), there were 4 colors (red, blue, yellow, and green) plus a colorless control. It was hypothesized that the liquid samples would be perceived as sweeter than the solid samples. The mean rating of the 5 liquid samples (7.61) was more that twice as high as the mean rating of the 5 solid samples (3.11). To determine whether this main effect for physical state held for each color, the mean difference in perceived sweetness between the liquid and solid samples by color was computed. A series of t tests revealed that the mean differences were significant at the .001 level in the expected direction for each color and the colorless control. There was no significant effect of color. These results strongly support the hypothesis that liquid samples are perceived as sweeter than solid samples. PMID- 9729848 TI - Which stress matters? The examination of temporal aspects of stress. AB - In this study, the impact of past experiences, present stressors, and expectations of future stress on psychological distress were explored. Participants were 38 male and 41 female spouses of patients hospitalized with non life-threatening diseases. Participants completed questionnaires on which they appraised past, present, and future stressors and described their levels of psychological distress. Five models of the relationships between appraisals of past, present, and future stressors with psychological distress were examined. Only one model was confirmed by the data. It suggests that an appraisal of past stressors affects the appraisal of present stressors only indirectly, through its effect on the appraisal of future stressors. This model also maintains that only the appraisal of present stressors affects psychological distress directly. The implications of these findings for stress research are discussed. PMID- 9729849 TI - Interocular suppression of a half-occluded region of stereoscopic images. AB - We assessed the detectability d' of a monocular small gray dot target presented on a half-occluded region of stereoscopic three-dimensional background images by comparing it with that on a two-dimensional (2D) region. For our experiments we used a typical two-alternative temporal forced-choice procedure, in which the target was presented in one of two temporal intervals for approximately 67 ms, and observers selected the interval they believed to have contained the target by pressing the corresponding key. To vary target signal intensity, we changed the target contrast against the background. According to signal-detection theory, we converted the percent-correct data to detectability d' and found that the relationship between d' and the contrast of the target followed Legge's equation. We used Legge's equation to calculate the contrast threshold and found that the contrast threshold of the target on the half-occluded region was higher than that on the 2D region. This elevation of contrast threshold indicates that interocular suppression of the half-occluded region occurs more frequently than that of the 2D region. We also refer to the monocular performance of the human visual system. PMID- 9729850 TI - Adaptive shift of visual sensitivity balance under ambient illuminant change. AB - We examined the relationship between the ambient illuminant chromaticity and changes in the sensitivity balance of the visual system, using illuminants of various chromaticities. The sensitivity of observers was measured in a room with a variable-chromaticity illuminant. The observer's state of chromatic adaptation was measured with unique-white settings. Our results showed that the change in visual sensitivity has a nonlinear correlation with the change in illuminant chromaticity; chromatic adaptation was nearly complete for desaturated illuminants, but the degree of chromatic adaptation became worse as the illuminant became more saturated. We defined a new index, relative cone weights, which represents this relationship well. To measure the role of chromatic induction from the immediate-surround area of the matching stimulus, we performed additional experiments by presenting the test inside a colored or black immediate surround. The results showed that the unique-white settings were not disturbed by the change in immediate-surround color. Our results imply that the room illuminant chromaticity was the primary factor in changing the observer's state of chromatic adaptation. PMID- 9729851 TI - Neural network approach for Compton-scattering imaging. AB - The problem of image reconstruction with Compton-scattering spectral data is an ill-posed problem, and the measurement error may be seriously amplified in the reconstruction result. For a stable solution, some kinds of a priori models of the problem should be incorporated into the process of reconstruction. Lee et al. [IEEE. Trans. Nucl. Sci. 40, 2049 (1993)] have proposed a continuous model with binary line processes. Owing to the coexistence of the continuous variable and the binary variable, the commonly used optimization methods for problems with continuous variables cannot be used in this case, and therefore a coupled gradient artificial neutral network was proposed for this mixed-integer problem. By introducing two interacting parts (with one part for the continuous variable and the other for the binary line processes) into the network, and by defining the appropriate energy function and dynamics, high-quality solutions were obtained upon convergence of the dynamics. Some simulated results are presented. PMID- 9729852 TI - Unified reconstruction theory for diffraction tomography, with consideration of noise control. AB - In diffraction tomography, the spatial distribution of the scattering object is reconstructed from the measured scattered data. For a scattering object that is illuminated with plane-wave radiation, under the condition of weak scattering one can invoke the Born (or the Rytov) approximation to linearize the equation for the scattered field (or the scattered phase) and derive a relationship between the scattered field (or the scattered phase) and the distribution of the scattering object. Reconstruction methods such as the Fourier domain interpolation methods and the filtered backpropagation method have been developed previously. However, the underlying relationship among and the noise properties of these methods are not evident. We introduce the concepts of ideal and modified sinograms. Analysis of the relationships between, and the noise properties of the two sinograms reveals infinite classes of methods for image reconstruction in diffraction tomography that include the previously proposed methods as special members. The methods in these classes are mathematically identical, but they respond to noise and numerical errors differently. PMID- 9729853 TI - Production and propagation of Hermite-sinusoidal-Gaussian laser beams. AB - Hermite-sinusoidal-Gaussian solutions to the wave equation have recently been obtained. In the limit of large Hermite-Gaussian beam size, the sinusoidal factors are dominant and reduce to the conventional modes of a rectangular waveguide. In the opposite limit the beams reduce to the familiar Hermite Gaussian form. The propagation of these beams is examined in detail, and resonators are designed that will produce them. As an example, a special resonator is designed to produce hyperbolic-sine-Gaussian beams. This ring resonator contains a hyperbolic-cosine-Gaussian apodized aperture. The beam mode has finite energy and is perturbation stable. PMID- 9729854 TI - Velocity difference measurement with a fiber-optic coupler. AB - Two single-mode fibers collect light with the same scattered wave vector from two spatially separated regions in a sample. These regions are illuminated by a single coherent laser beam, so that the collected signals interfere when combined by means of a fiber-optic coupler, before they are directed to a photomultiplier tube. The fibers and the coupler are polarization preserving to guarantee a high signal-to-noise ratio. The measured intensity fluctuations are used to determine the velocity difference omega v(L) for spatial separations L in the sample. Specifically, an intensity autocorrelation function is calculated theoretically for rigid body rotation and is tested experimentally. Experimental results span two orders of magnitude in L and agree with theoretical predictions with an error of less than 5%. This new technique will be very useful in the study of turbulent flow and particle settling dynamics. PMID- 9729855 TI - Psychophysical signatures associated with magnocellular and parvocellular pathway contrast gain: comment. AB - The recent paper by Pokorny and Smith [J. Opt. Soc. Am. A 14, 2477 (1977)] describes psychophysical data that correlate well with the physiology of the magnocellular and the parvocellular pathways. One of the critical differences between cells in these two pathways is the contrast of the visual input for which the responses of these cells start to saturate. In the analysis of their data, Pokorny and Smith have to assume values for the saturation contrast of the putative pathways that mediate detection. Here I reanalyze their model and show that this assumption is unnecessary, since values of the saturation contrast follow directly from their data. PMID- 9729856 TI - Measurement of the wave-front aberration of the eye by a fast psychophysical procedure. AB - We used a fast psychophysical procedure to determine the wave-front aberrations of the human eye in vivo. We measured the angular deviation of light rays entering the eye at different pupillary locations by aligning an image of a point source entering the pupil at different locations to the image of a fixation cross entering the pupil at a fixed location. We fitted the data to a Zernike series to reconstruct the wave-front aberrations of the pupil. With this technique the repeatability of the measurement of the individual coefficients was 0.019 micron. The standard deviation of the overall wave-height estimation across the pupil is less than 0.3 micron. Since this technique does not require the administration of pharmacological agents to dilate the pupil, we were able to measure the changes in the aberrations of the eye during accommodation. We found that administration of even a mild dilating agent causes a change in the aberration structure of the eye. PMID- 9729857 TI - Comparison of the eye's wave-front aberration measured psychophysically and with the Shack-Hartmann wave-front sensor. AB - The Shack-Hartmann wave-front sensor offers many theoretical advantages over other methods for measuring aberrations of the eye; therefore it is essential that its accuracy be thoroughly tested. We assessed the accuracy of a Shack Hartmann sensor by directly comparing its measured wave-front aberration function with that obtained by the Smirnov psychophysical method for the same eyes. Wave front profiles measured by the two methods agreed closely in terms of shape and magnitude with rms differences of approximately lambda/2 and approximately lambda/6 (5.6-mm pupil) for two eyes. Primary spherical aberration was dominant in these profiles, and, in one subject, secondary coma was opposite in sign to primary coma, thereby canceling its effect. Discovery of an unusual, subtle wave front anomaly in one individual further demonstrated the accuracy and sensitivity of the Shack-Hartmann wave-front sensor for measuring the optical quality of the human eye. PMID- 9729858 TI - Estimates of the ocular wave aberration from pairs of double-pass retinal images. AB - We apply a computational technique to retrieve the wave aberration of the eye from the point-spread function obtained from pairs of double-pass retinal images. The method consists of an adapted pyramidal version of a nonlinear least-squares fitting procedure to a wave aberration expressed as an expansion in Zernike polynomials. Although the procedure provides accurate estimates of the wave aberration, it presents several drawbacks that are discussed in detail. In particular, since a great deal of computational time is necessary to retrieve a single wave aberration, this technique is not useful for real-time applications. We present results of wave aberrations in five normal subjects in the fovea for a 4-mm-pupil diameter. In every case there is a clear presence of comalike aberrations, while the third-order spherical aberration is usually smaller than previous estimates. The root-mean-square error in the retrieved wave aberration, when defocus and astigmatism were corrected, ranges from 0.24 to 0.5 wavelength. The particular values of the aberration coefficients present a large intersubject variability. PMID- 9729859 TI - Assessment of the accuracy of the crossed-cylinder aberroscope technique. AB - Simulations of the optics of the Howland crossed-cylinder aberroscope technique show that errors in alignment, data collection, and analysis can lead to unexpected asymmetries of the determined aberrations in a rotationally symmetric system. In particular, coma can be incorrectly indicated. The magnitude of the error in aberration measurement depends on the magnitude of the alignment, data collection, and alignment errors. These findings indicate that the tolerances for setting up the technique and data collection should be analyzed thoroughly before quantitative significance is given to the determined aberration coefficients. PMID- 9729860 TI - Choice of reference axis in ocular wave-front aberration measurement. AB - The geometrical center of the pupil has often been used as the reference axis in ocular wave-front aberration measurement. However, the geometrical center of the pupil may shift when the pupil size changes under different conditions. In particular, for subjective methods, defining the center of the pupil precisely during the actual measurement is not always practical. Furthermore, the geometrical center of the pupil may not define the chief ray of the ocular optics because of the Stiles-Crawford apodization effect, which has a peak location that often deviates from the geometrical center of the pupil. We present the coefficient transformation table of the Taylor polynomial up to the sixth order with respect to reference axis shift. We illustrate the effect of wave-front aberration change with reference axis shift with experimental data. This type of wave-front aberration change could be a true measurement error if there is an error in defining the reference axis. We also propose using the coaxially sighted corneal reflex as a better reference axis in aberration measurement. PMID- 9729861 TI - Lie algebraic treatment of dioptric power and optical aberrations. AB - The dioptric power of an optical system can be expressed as a four-component dioptric power matrix. We generalize and reformulate the standard matrix approach by utilizing the methods of Lie algebra. This generalization helps one deal with nonlinear problems (such as aberrations) and further extends the standard matrix formulation. Explicit formulas giving the relationship between the incident and the emergent rays are presented. Examples include the general case of thick and thin lenses. The treatment of a graded-index medium is outlined. PMID- 9729862 TI - Foveal optical modulation transfer function of the human eye at various pupil sizes. AB - We determined the foveal optical modulation transfer functions of the human eye (O) for pupil sizes of 1-8 mm by using two simple psychophysical techniques. O as a function of spatial frequency f could be described by exp[-(f/fc)n] at any pupil size in our data as well as in the data available in the literature [J. Physiol. (London) 186, 558 (1966); Opt. Acta 21, 395 (1974); Vision Res. 33, 15 (1993); J. Opt. Soc. Am. A 11, 246 (1994)]. When all these estimates of fc and n were pooled the parameters were found to depend on the pupil diameter as fc = 16.6 - 1.49p and n = exp(0.840/p - 0.318). This result indicates that at 1 mm O(f) is close to the diffraction-limited system. PMID- 9729863 TI - Influence of amount and changes in axis of astigmatism on retinal image quality. AB - We measured retinal image quality in astigmatic eyes, using the double-pass technique. We analyzed the influence of the amount of astigmatism and changes in axis of astigmatism on the eye's optical performance. Different amounts of astigmatism were obtained by variation of the cylindrical power of a lens situated in front of the eye, between 0.25-diopter (D) overcorrection and 1-D undercorrection at intervals of 0.25 D. Changes in the axis of astigmatism were obtained by rotation of the lens, which neutralizes the astigmatism in an angle of +/- 10 degrees at 5 degrees intervals. The results show the decrease in retinal image quality and the increase in the degree of image astigmatism obtained when the amount of astigmatism increases or the angle between the lens and the eye axis is other than zero. In general, the largest variations correspond to when the astigmatism changes from 0 to 0.25 D or when the axis changes from 0 degree to +/- 5 degrees. The reduction in optical performance is smaller in living eyes than in an eye model or in an artificial eye. The aberrations present in the living eye reduce the relative loss of retinal image quality introduced by astigmatism. PMID- 9729865 TI - Optical isolation of portions of a wave front. AB - A criterion is established for determining when portions of a wave front can be said to be optically isolated from the rest of the wave front in the sense that they can subsequently be treated separately when one is considering the formation of images. The subarea of the wave front is treated as a separate aperture, and it is said to be isolated if diffraction maxima for the majority of the wave front fall at or beyond the first minima for the subarea. An illustrative example employing two circular unequal-diameter apertures is presented. A method is given for identifying portions of wave front that may be optically isolated; the method uses the technique of fitting a reference surface to the actual wave front and then finding what is defined as the differential deflection of the actual surface with respect to the reference surface at all locations. Subpopulations of locations with similar differential deflection values, sufficient numbers, and sufficient differential deflection are candidates for area of optical isolation. PMID- 9729864 TI - Monochromatic aberrations and point-spread functions of the human eye across the visual field. AB - The monochromatic aberrations of the human eye along the temporal meridian are studied by a novel laser ray-tracing method. It consists of delivering a narrow laser pencil into the eye through a given point on the pupil and recording the aerial image of the retinal spot with a CCD camera. The relative displacement of this image is proportional to the geometrical aberration of the ray (laser pencil) at the retina. We scanned the pupils of four observers in steps of 1 mm (effective diameter, 6.7 mm) and for five field angles (0 degree, 5 degrees, 10 degrees, 20 degrees, and 40 degrees). In addition, the aerial image for each chief ray is a low-pass-filtered version of the retinal point-spread function corresponding to a fully dilated pupil. The resulting spot diagrams, displaying the distribution of ray aberrations, are highly correlated with these point spread functions. We have estimated the wave-front error by fitting Zernike polynomials (up to the fifth order). Despite the large variation found among observers, the overall rms wave-front error is relatively homogeneous. At the fovea, the average rms value was 1.49 microns when the second-order terms (defocus and astigmatism) were considered; this was reduced to 0.45 micron when the second-order terms were ignored. The rms values increase slowly, in a roughly linear fashion with eccentricity, such that at 40 degrees they are approximately double. These results are consistent with previous findings on the off-axis optical quality of the eye. PMID- 9729866 TI - Predicting the effects of optical defocus on human contrast sensitivity. AB - We used diffraction modulation transfer functions and model eyes to predict the effect of defocus on the contrast sensitivity function (CSF) and compared these predictions with previously published experimental data. Using the principle that optically induced changes in the modulation transfer function should be paralleled by identical changes in the CSF, we used the modulation transfer function calculations with the best-focus CSF measurements to predict the defocused CSF. An aberration-free model predicted the effects of defocus well when the CSF was measured with small pupils (e.g., 2 mm) but not with larger pupils (6-8 mm). When the model included average aberrations, prediction of the defocused CSF with large pupils was better but remained inaccurate, failing, in particular, to reflect differences between individual subjects. Inclusion of measured aberrations for individual subjects provided accurate predictions in the shape of the monochromatic CSF of two of three subjects with hyperopic defocus and good predictions of the polychromatic CSF of two subjects with hyperopic defocus. Prediction of the effects of myopic defocus by use of measured individual aberrations of one subject were less successful. Hence a diffraction optics model can provide good predictions of the effects of defocus on the human CSF, given that one has knowledge of the individual ocular aberrations. These predictions are dependent on the quality of the aberration measurements. PMID- 9729867 TI - Influence of Stiles-Crawford effect apodization on spatial visual performance. AB - The Stiles-Crawford effect is often invoked by vision scientists when predictions of the effects of aberrations and defocus on spatial visual performance are not borne out experimentally. Modeling the Stiles-Crawford effect as an apodization, we investigated the expected influence that it would have on spatial visual performance in the presence of 1-diopter primary spherical aberration at the edge of a 6-mm-diameter centered pupil. The changes in refraction produced by a high Stiles-Crawford effect, according to various criteria, were small at approximately 0.10 diopter. The Stiles-Crawford effect has only a small capability to compensate for defocus and spherical aberration. These results indicate that the Stiles-Crawford effect has little influence on spatial visual performance in the case of centered pupils. We suggest that the faith that has often been placed in the Stiles-Crawford effect to account for discrepancies between experimental results and expected results is not justified, at least for well-centered pupils and Stiles-Crawford effects. PMID- 9729868 TI - Correction of the aberrations in the human eye with a liquid-crystal spatial light modulator: limits to performance. AB - We evaluated the performance of a liquid-crystal spatial light modulator for static correction of the aberrations in the human eye. By applying phase retrieval techniques to pairs of double-pass images we first estimated the wave aberration of the eye to be corrected. Then we introduced the opposite phase map in the modulator, which was placed in a plane conjugated with the eye's pupil, and we recorded double-pass images of a point source before and after correction of the aberrations. In a slightly aberrated artificial eye a clear improvement was obtained after correction, and, although diffraction-limited performance was not achieved, the results were close to the theoretical predictions. In the two living eyes that we studied some benefit also appeared in the correction, but the performance was worse than that expected. We evaluated possible explanations for the relatively poor performance that was obtained in the human eye: an incorrect estimate of the ocular aberration, the limited spatial resolution of the modulator, and the dynamic changes in the ocular aberrations. Based on the results in the artificial eye, the first problem was not considered to be a major source of error. However, we showed that the spatial resolution of the liquid crystal spatial light modulator limits the maximum correction to be attained. In addition, the changes in the ocular optics over time also impose a limit in the performance of static corrections. PMID- 9729869 TI - Generation of third-order spherical and coma aberrations by use of radically symmetrical fourth-order lenses. AB - We have extended the method of Alvarez [J. Am. Optom. Assoc. 49, 24 (1978)] to generate a variable magnitude of third-order spherical and/or coma aberration by using a combination of fourth-order plates with a magnification system. The technique, based on the crossed-cylinder aberroscope, is used to measure the wave front aberration generated by the plates. The method has been applied to correct the third-order spherical aberration generated by an artificial eye as well as the coma produced by a progressive addition ophthalmic lens. The simplicity of the method and its relatively low cost make it attractive for partial correction of the aberrations of the eye. PMID- 9729871 TI - Optimal corneal ablation for eyes with arbitrary Hartmann-Shack aberrations. AB - New technologies for accurately measuring corneal shape and full eye aberrations are now available. An algorithm that uses these technologies to predict the amount of ablation needed to produce a corneal surface that optimally focuses light is developed. It is found that knowledge of the aberrations is far more important than knowledge of corneal shape. Neglect of corneal shape information introduces an error of less than approximately 0.05 micron in the optimal ablation depth. Neglect of the aberrations is a different story. Small changes in the aberration structure, such as going from the optimal ablation to a spherical ablation, introduce ablation changes of greater than 10 microns. It is argued that there are many occasions when less ablation can lead to improved image quality. PMID- 9729870 TI - Custom photorefractive keratectomy ablations for the correction of spherical and cylindrical refractive error and higher-order aberration. AB - Photorefractive keratectomy is an evolving refractive procedure for correcting myopia, hyperopia, and astigmatism. Earlier descriptions of the patterns required for this surgery are based on paraxial optics. In this investigation the required pattern is generalized to account for spherical refractive error (defocus), axial astigmatism of arbitrary orientation, and fourth-order aberrations of the eye. The patterns described in this study can be used to customize photorefractive keratectomy and to provide corrections that account for aberration content as well as paraxial values. Furthermore, a description of the pattern along the boundary of the optical zone is given, which may prove useful in designing blending zones. An example of the use of these techniques is given for a schematic eye model. PMID- 9729873 TI - Sensitivity of the relative-rate test to taxonomic sampling. AB - Relative-rate tests may be used to compare substitution rates between more than two sequences, which yields two main questions: What influence does the number of sequences have on relative-rate tests and what is the influence of the sampling strategy as characterized by the phylogenetic relationships between sequences? Using both simulations and analysis of real data from murids (APRT and LCAT nuclear genes), we show that comparing large numbers of species significantly improves the power of the test. This effect is stronger if species are more distantly related. On the other hand, it appears to be less rewarding to increase outgroup sampling than to use the single nearest outgroup sequence. Rates may be compared between paraphyletic ingroups and using paraphyletic outgroups, but unbalanced taxonomic sampling can bias the test. We present a simple phylogenetic weighting scheme which takes taxonomic sampling into account and significantly improves the relative-rate test in cases of unbalanced sampling. The answers are thus: (1) large taxonomic sampling of compared groups improves relative-rate tests, (2) sampling many outgroups does not bring significant improvement, (3) the only constraint on sampling strategy is that the outgroup be valid, and (4) results are more accurate when phylogenetic relationships between the investigated sequences are taken into account. Given current limitations of the maximum-likelihood and nonparametric approaches, the relative-rate test generalized to any number of species with phylogenetic weighting appears to be the most general test available to compare rates between lineages. PMID- 9729872 TI - Influence of myopia and aging on the optimal spherical aberration of soft contact lenses. AB - Soft contact lenses with different levels of third-order spherical aberration were tested in two samples of subjects aged between 20 and 45 years: 18 emmetropes and 19 myopes. Contrast sensitivity was measured at 12 cycles/degree to determine the optimal lens spherical aberration required by each individual. The optimal third-order coefficient was found to be negative on average in both refractive error groups. Myopic subjects required contact lenses with more negative spherical aberration than did emmetropes. The optimal aberration was also found to become increasingly negative with aging. The rate of this age related change was faster in the myopic group. In comparison with aberration-free soft contact lenses, an improvement in contrast detection threshold of more than 25% was observed with optimal spherical aberration in half of the myopic subjects. PMID- 9729874 TI - Evolution of the "classical" cadherin family of cell adhesion molecules in vertebrates. AB - The cadherins are major mediators of calcium-dependent cell-cell adhesion and are also involved in cell signaling pathways during development. The classical cadherins, which are the definitive group of the cadherin superfamily, are transmembrane proteins that consist of an extracellular domain of five cadherin repeats, including an HAV tripeptide conserved in one binding surface within the first domain, and a highly conserved cytoplasmic domain that interacts with the actin cytoskeleton via the catenin proteins. These cadherins play major roles in vertebrate morphogenesis; they are expressed widely throughout development, antibodies to specific cadherins perturb a variety of developmental processes, and many gene knockouts are lethal at early stages of development. Phylogenetic analysis of the "classical" cadherins shows that in the vertebrates there are four paralog families. The rate of evolutionary change is radically different between the different paralogs, indicating that there are significantly different selection pressures on the functions of the various cadherins, both between the different paralogs in a single organism lineage and between different organism lineages within a single paralog family. There is also evidence for gene conversion between the E-cadherin and P-cadherin paralogs in Gallus gallus and possibly Xenopus laevis, but not between the same paralogs in the mammalian lineages. A scheme for the origin of the paralogs within the vertebrate lineage based on these analyses indicates that the presence of the four paralog families is a characteristic of vertebrates and that variation of cadherin structure and function is a significant factor in morphological evolution of vertebrates. PMID- 9729875 TI - Phylogenetic resolution of complex mutational features at Y-STR DYS390 in aboriginal Australians and Papuans. AB - Y-chromosomal short tandem repeats (STRs) are used for the study of male aspects of human evolution as well as for forensic applications and paternity testing. Both applications require an understanding of the underlying mutational mechanisms that create variability. We describe complex mutations at the substructured DYS390 STR locus in 97 natives of the New Guinea/Australian region. Sequencing of short alleles in these populations indicates multirepeat deletions. All samples are further characterized using the five additional Y-STR loci DYS19, DXYS156-Y, DYS391, DYS392, and DYS393. Phylogenetic analysis of the resulting haplotypes yields ethnically specific clusters predating the settlement of Australia and Papua New Guinea (although archaic Homo sapiens or Homo erectus lineages are absent). The phylogeny confirms that DYS390 violates the stepwise mutation model and demonstrates that the DYS390 locus mutates relatively rapidly and retains its variability after structural change. PMID- 9729876 TI - A new AluI satellite DNA in the root-knot nematode Meloidogyne fallax: relationships with satellites from the sympatric species M. hapla and M. chitwoodi. AB - A highly abundant satellite DNA comprising 20% of the Meloidogyne fallax (Nematoda, Tylenchida) genome was cloned and sequenced. The satellite monomer is 173 bp long and has a high A + T content of 72.3%, with frequent runs of A's and T's. The sequence variability of the monomers is 2.7%, mainly due to random distribution of single-point mutations. A search for evidence of internal repeated subunits in the monomer sequence revealed a 6-bp motif (AAATTT) for which five degenerated repeats, differing by just a single base pair, could be identified. Pairwise comparison of the M. fallax satellite with those from the sympatric species Meloidogyne chitwoodi and Meloidogyne hapla revealed a high sequence similarity (68.39%) with one satellite DNA subfamily in M. chitwoodi, which indicated an unexpected close relationship between them. Given the high copy number and the extreme sequence homogeneity among monomeric units, it may be assumed that the satellite DNA of M. fallax could have evolved through some recent and extensive amplification burst in the nematode genome. In this case, its relatively short life would not yet have allowed the accumulation of random mutations in independent amplified repeats. Considering the morphological resemblance between the two species and their ability to produce interspecific fertile hybrids under controlled conditions, these results indicate that M. fallax may share a common ancestor with M. chitwoodi, from which it could have diverged recently. All these data suggest that M. fallax could be the result of a recent speciation process and show that Meloidogyne satellite DNAs may be of interest to resolve phylogenetic relationships among closely related species from this genus. PMID- 9729877 TI - The RTE class of non-LTR retrotransposons is widely distributed in animals and is the origin of many SINEs. AB - RTE-1 is a non-long-terminal-repeat (non-LTR) retrotransposable element first found in the Caenorhabditis elegans genome. It encodes a 1,024-amino-acid open reading frame (ORF) containing both apurinic-apyrimidic endonuclease and reverse transcriptase domains. A possible first ORF of only 43 amino acids overlaps with the larger ORF and may be the site of translation initiation. Database searches and phylogenetic analysis indicate that representatives of the RTE clade of non LTR retrotransposons are found in the bovine and sheep genomes of mammals and in the silkmoth and mosquito genomes of insects. In addition, the previously identified SINEs, Art2 and Pst, from ruminate and viper genomes are shown to be truncated RTE-like retrotransposable elements. RTE-derived SINE elements are also found in mollusc and flatworm genomes. Members of the RTE clade are characterized by unusually short 3' untranslated regions that are predominantly composed of AT rich trimer, tetramer, and/or pentamer repeats. This study establishes RTE as a very widespread clade of non-LTR retrotransposons. RTE represents the third distinct class of non-LTR retrotransposons in the vertebrate lineage (after Line 1 elements in mammals and CR1 elements in birds and reptiles). PMID- 9729878 TI - The core domain of retrotransposon integrase in Hordeum: predicted structure and evolution. AB - Propagation of long terminal repeat (LTR)-bearing retrotransposons and retroviruses requires integrase (IN, EC 2.7.7.-), encoded by the retroelements themselves, which mediates the insertion of cDNA copies back into the genome. An active retrotransposon family, BARE-1, comprises approximately 7% of the barley (Hordeum vulgare subsp. vulgare) genome. We have generated models for the secondary and tertiary structure of BARE-1 IN and demonstrate their similarity to structures for human immunodeficiency virus 1 and avian sarcoma virus INs. The IN core domains were compared for 80 clones from 28 Hordeum accessions representative of the diversity of the genus. Based on the structural model, variations in the predicted, aligned translations from these clones would have minimal structural and functional effects on the encoded enzymes. This indicates that Hordeum retrotransposon IN has been under purifying selection to maintain a structure typical of retroviral INs. These represent the first such analyses for plant INs. PMID- 9729879 TI - Ancient large-scale genome duplications: phylogenetic and linkage analyses shed light on chordate genome evolution. AB - Paralogous genes from several families were found in four human chromosome regions (4p16, 5q33-35, 8p12-21, and 10q24-26), suggesting that their common ancestral region underwent several rounds of large-scale duplication. Searches in the EMBL databases, followed by phylogenetic analyses, showed that cognates (orthologs) of human duplicated genes can be found in other vertebrates, including bony fishes. In contrast, within each family, only one gene showing the same high degree of similarity with all the duplicated mammalian genes was found in nonvertebrates (echinoderms, insects, nematodes). This indicates that large scale duplications occurred after the echinoderms/chordates split and before the bony vertebrate radiation. It has been suggested that two rounds of gene duplication occurred in the vertebrate lineage after the separation of Amphioxus and craniate (vertebrates + Myxini) ancestors. Before these duplications, the genes that have led to the families of paralogous genes in vertebrates must have been physically linked in the craniate ancestor. Linkage of some of these genes can be found in the Drosophila melanogaster and Caenorhabditis elegans genomes, suggesting that they were linked in the triploblast Metazoa ancestor. PMID- 9729880 TI - Evolution of the copia retrotransposon in the Drosophila melanogaster species subgroup. AB - We report the results of a phylogenetic survey of the retrotransposon copia in the melanogaster subgroup of the Drosophila genus. The polymerase chain reaction was used to amplify the copia 5' long terminal repeat and the adjacent untranslated leader region from representative melanogaster subgroup species. Restriction and sequence analyses of this region reveal discrete classes of copia size variants within the melanogaster subgroup. Phylogenetic comparisons of copia sequence data indicate that the size variants represent different copia subfamilies which diverged prior to their distribution in the melanogaster subgroup. Our results also suggest that copia elements have been subject to horizontal and vertical transmission during their evolution. PMID- 9729882 TI - A covariotide model explains apparent phylogenetic structure of oxygenic photosynthetic lineages. AB - The aims of the work were (1) to develop statistical tests to identify whether substitution takes place under a covariotide model in sequences used for phylogenetic inference and (2) to determine the influence of covariotide substitution on phylogenetic trees inferred for photosynthetic and other organisms. (Covariotide and covarion models are ones in which sites that are variable in some parts of the underlying tree are invariable in others and vice versa.) Two tests were developed. The first was a contingency test, and the second was an inequality test comparing the expected number of variable sites in two groups with the observed number. Application of these tests to 16S rDNA and tufA sequences from a range of nonphotosynthetic prokaryotes and oxygenic photosynthetic prokaryotes and eukaryotes suggests the occurrence of a covariotide mechanism. The degree of support for partitioning of taxa in reconstructed trees involving these organisms was determined in the presence or absence of sites showing particular substitution patterns. This analysis showed that the support for splits between (1) photosynthetic eukaryotes and prokaryotes and (2) photosynthetic and nonphotosynthetic organisms could be accounted for by patterns arising from covariotide substitution. We show that the additional problem of compositional bias in sequence data needs to be considered in the context of patterns of covariotide/covarion substitution. We argue that while covariotide or covarion substitution may give rise to phylogenetically informative patterns in sequence data, this may not always be so. PMID- 9729881 TI - Evolution and phylogenetic utility of the period gene in Lepidoptera. AB - Evolution and phylogenetic utility of the period gene are explored through sequence analysis of a relatively conserved 909-bp fragment in 26 lepidopteran species. Taxa range from tribes to superfamilies, primarily within the putative clade Macrolepidotera plus near outgroups, and include both strongly established and problematic groupings. Their divergence dates probably range from the late Cretaceous through much of the Tertiary. Comparisons within the same set of closely related species show that amino acid substitutions in period occur 4.9 and 44 times as frequently as they do in two other nuclear genes--dopa decarboxylase and elongation factor-1 alpha, respectively. In contrast, rates of observed synonymous substitution are within 60% of each other for these three genes. Synonymous changes in period approach saturation by the family level, whereas nonsynonymous and amino acid divergences across the Macrolepidoptera are less than half the maximal values reported for this gene. Phylogenetic analyses of period strongly supported groupings at the family level and below. In contrast to previous analyses at this level with other nuclear genes, much of the information lies in nonsynonymous change. Relationships up to the superfamily level were recovered with decreasing effectiveness, and little, if any, signal was apparent regarding relationships among superfamilies. This could reflect rapid radiation of the superfamilies, however, rather than saturation in the period locus; thus, period, in combination with other genes, remains a plausible candidate for approaching the difficult problems of lepidopteran family and superfamily relationships. PMID- 9729883 TI - Evolutionary history of free-swimming and sessile lifestyles in urochordates as deduced from 18S rDNA molecular phylogeny. AB - Whether the ancestral chordates were free-swimming or sessile is a longstanding question that remains to be settled. Vertebrates and amphioxi are free-swimming, but the most basal chordate subphylum (the urochordates) includes both sessile and free-swimming species. Here, 1 report molecular phylogenetic analyses of 18S rDNA of urochordates to deduce which lifestyle is ancestral. This revealed a close relationship between salps and doliolids and paraphyly of the ascidians. An early divergence of larvaceans, which show a tadpole-like body plan throughout life, is also supported by the analyses. Based on this phylogeny, a free-swimming ancestor for chordates is more parsimonious than a sessile ancestor. The evolutionary history of various lifestyles of chordates from this ancestral form is proposed. PMID- 9729884 TI - Evolution of anaerobic ciliates from the gastrointestinal tract: phylogenetic analysis of the ribosomal repeat from Nyctotherus ovalis and its relatives. AB - The 18S and 5.8S rDNA genes and the internal transcribed spacers ITS-1 and ITS-2 of ciliates living in the hindgut of frogs, millipedes, and cockroaches were analyzed in order to study the evolution of intestinal protists. All ciliates studied here belong to the genus Nycrotherus. Phylogenetic analysis revealed that these ciliates from a monophyletic group that includes the distantly related anaerobic free-living heterotrichous ciliates Metopus palaeformis and Metopus contortus. The intestinal ciliates from the different vertebrate and invertebrate hosts are clearly divergent at the level of their rDNA repeats. This argues for the antiquity of the associations and a predominantly vertical transmission. This mode of transmission seems to be controlled primarily by the behavior of the host. The different degrees of divergence between ciliates living in different strains of one and the same cockroach species most likely reflect the different geographical origins of the hosts. In addition, host switches must have occurred during the evolution of cockroaches, since identical ciliates were found only in distantly related hosts. These phenomena prevent the reconstruction of potential cospeciation events. PMID- 9729885 TI - Evidence of independent gene duplications during the evolution of archaeal and eukaryotic family B DNA polymerases. AB - Eukaryotes and archaea both possess multiple genes coding for family B DNA polymerases. In animals and fungi, three family B DNA polymerases, alpha, delta, and epsilon, are responsible for replication of nuclear DNA. We used a PCR-based approach to amplify and sequence phylogenetically conserved regions of these three DNA polymerases from Giardia intestinalis and Trichomonas vaginalis, representatives of early-diverging eukaryotic lineages. Phylogenetic analysis of eukaryotic and archaeal paralogs suggests that the gene duplications that gave rise to the three replicative paralogs occurred before the divergence of the earliest eukaryotic lineages, and that all eukaryotes are likely to possess these paralogs. One eukaryotic paralog, epsilon, consistently branches within archaeal sequences to the exclusion of other eukaryotic paralogs, suggesting that an epsilon-like family B DNA polymerase was ancestral to both archaea and eukaryotes. Because crenarchaeote and euryarchaeote paralogs do not form monophyletic groups in phylogenetic analysis, it is possible that archaeal family B paralogs themselves evolved by a series of gene duplications independent of the gene duplications that gave rise to eukaryotic paralogs. PMID- 9729886 TI - Characterization of AmphiF-spondin reveals the modular evolution of chordate F spondin genes. AB - The F-spondin genes are a family of extracellular matrix molecules united by two conserved domains, FS1 and FS2, at the amino terminus plus a variable number of thrombospondin repeats at the carboxy terminus. Currently, characterized members include a single gene in Drosophila and multiple genes in vertebrates. The vertebrate genes are expressed in the midline of the developing embryo, primarily in the floor plate of the neural tube. To investigate the evolution of chordate F spondin genes, I have used the basal position in chordate phylogeny of the acraniate amphioxus. A single F-spondin-related gene, named AmphiF-spondin, was isolated from amphioxus. Based on molecular phylogenetics, AmphiF-spondin is closely related to a particular subgroup of vertebrate F-spondin genes that encode six thrombospondin repeats. However, unlike these genes, expression of AmphiF-spondin is not confined to the midline but is found through most of the central nervous system. Additionally, AmphiF-spondin has lost three thrombospondin repeats and gained two fibronectin type III repeats, one of which has strong identity to a fibronectin type III repeat from Deleted in Colorectal Cancer (DCC). Taken together, these results suggest a complex evolutionary history for chordate F-spondin genes that includes (1) domain loss, (2) domain gain by tandem duplication and divergence of existing domains, and (3) gain of heterologous domains by exon shuffling. PMID- 9729887 TI - Optimally recovering rate variation information from genomes and sequences: pattern filtering. AB - Nucleotide substitution rates vary at different positions within genes and genomes, but rates are difficult to estimate, because they are masked by the stochastic nature of substitutions. In this paper, a linear method, pattern filtering, is described which can optimally separate the signals (related to substitution rates or to other measures of sequence change) from stochastic noise. Pattern filtering promises to be useful in both genomic and molecular evolution studies. In an example using mitochondrial genomes, it is shown that pattern filtering can reveal coding and non-coding regions without the need for prior identification of reading frames or other knowledge of the sequence and promises to be an important tool for genomic analysis. In a second example, it is shown that pattern filtering allows one to classify sites on the basis of an estimator of substitution rates. Using elongation factor EF-1 alpha sequences, it is shown that the fastest sites favor archaea as the sister taxon of eukaryotes, whereas the slower sites support the eocyte prokaryotes as the sister taxon of eukaryotes, suggesting that the former result is an artifact of "long branch attraction." PMID- 9729888 TI - A book of life? How the genome became an information system and DNA a language. PMID- 9729889 TI - The Olmec heart effigy revisited: a highly accurate, ancient depiction of the human heart. PMID- 9729891 TI - Historical and recent evidence for the existence of mitogenetic radiation. PMID- 9729892 TI - Cytosolic beta-cyanoalanine synthase activity attributed to cysteine synthases in cocklebur seeds. Purification and characterization of cytosolic cysteine synthases. AB - The activity of beta-cyanoalanine synthase (CAS, EC 4.4.1.9) in cotyledons of cocklebur seeds (Xanthium pennsylvanicum Wallr.) was detected both in the soluble and particulate fractions. The CAS activity of the soluble fraction (cytosolic CAS activity) was 10 times higher than that of the particulate fraction. The CAS activity of the particulate fraction was confirmed to be localized in the mitochondria. Both enzymatic activities were clearly separated by non-denaturing PAGE. The enzyme with cytosolic CAS activity has been extensively purified and separated into three different forms designated as cyt-1, cyt-2, and cyt-3. According to the SDS-PAGE analysis, the three enzymes are estimated to be a homodimer composed of 35-kDa subunits. The purified enzymes showed CS activity. Partial amino acid sequences of cyt-1 were determined and had a high homology with cysteine synthases (CS, EC 4.2.99.8) from other plant sources. The catalytic action of the purified CSs in converting cyanide and cysteine into H2S and beta cyanoalanine was confirmed by the detection of significant 14CN incorporation into beta-cyanoalanine. These results indicated that cytosolic CAS activity is due to cytosolic CS and suggested that the CAS activity of CS is likely to be involved in cyanide metabolism in plant tissues. PMID- 9729893 TI - The expression pattern of the gene for NPK1 protein kinase related to mitogen activated protein kinase kinase kinase (MAPKKK) in a tobacco plant: correlation with cell proliferation. AB - Mitogen-activated protein kinase (MAPK) cascades consist of members of three families of protein kinases: the MAPK family, the MAPK kinase family, and the MAPK kinase kinase (MAPKKK) family. Some of these cascades have been shown to play central roles in the transmission of signals that control various cellular processes including cell proliferation. Protein kinase NPK1 is a structural and functional tobacco homologue of MAPKKK, but its physiological function is yet unknown. In the present study, we have investigated sites of expression of the NPK1 gene in a tobacco plant and developmental and physiological controls of this expression. After germination, expression of NPK1 was first detected in tips of a radicle and cotyledons, then in shoot and root apical meristems, surrounding tissues of the apical meristems, primordia of lateral roots, and young developing organs. No expression was, however, observed in mature organs. Incubation of discs from mature leaves of tobacco with both auxin and cytokinin induced NPK1 expression before the division of cells. It was also induced at early stages of the development of primordia of lateral roots and adventitious roots. Thus, NPK1 expression appears to be tightly correlated with cell division or division competence. Even when an inhibitor of DNA synthesis was added during the germination or the induction of lateral roots by auxin, NPK1 expression was detected. These results showed that the NPK1 expression precedes DNA replication. We propose that NPK1 participates in a process involving the division of plant cells. PMID- 9729894 TI - Expression of multiple AGAMOUS-like genes in male and female flowers of cucumber (Cucumis sativus L.). AB - Members of the MADS-box gene family control reproductive development in higher plants. In cucumber, floral development exhibits several interesting features related to a genetically determined sex-expression mechanism, that affects the differentiation of male and female flowers. In this study, three cDNA homologues of the homeotic gene AGAMOUS have been cloned from early-stage floral buds of Cucumis sativus and fully sequenced. Their expression was studied by Northern analysis using two contrasting sex genotypes, an androecious line and a gynoecious one. The three genes are expressed at low levels at earlier bud stages, the levels rising as the bud matures. Two of the clones, CAG1 and CAG3, are expressed in the third and fourth whorl of mature flowers, while CAG2 is restricted to the carpel; none is expressed in leaves. The transcript levels do not appear to be modulated by gibberellin or ethephon, two treatments that alter sex expression in cucumber. While MADS-box genes probably play an essential role in cucumber floral development, as they do in other plants, our findings may imply that the pathway leading to reproductive organ arrest in cucumber unisexual buds acts independently of MADS-box gene expression. PMID- 9729895 TI - The identification of DNA binding factors specific for as-1-like sequences in auxin-responsive regions of parA, parB and parC. AB - We have identified the auxin-responsive region (Aux-RR) of the parA promoter; it is derived from a gene that is induced by auxin in tobacco mesophyll protoplasts. By analyses of gain-of-function and point mutations in transgenic tobacco plants, we showed that an as-1-like sequence was required, but not alone sufficient, for auxin responsiveness of the parA promoter, as has been also shown for the parC promoter in a recent study. A gel mobility shift assay revealed that the as-1 like sequence of the parA promoter bound specifically to the as-1 binding nuclear factor, ASF-1, while the as-1-like sequence of the parC promoter bound specifically to a novel nuclear factor named ALF-1 (as-1-like sequence binding factor-1). Since these bindings were correlated with auxin-induced activation of transcription, those results suggest that ALF-1 is a new nuclear factor involved in auxin responsiveness and that it is distinct from ASF-1. Furthermore, we found that an as-1-like sequence in the AuxRR of parB bound specifically to a tobacco nuclear factor, named ALF-2, which differed from ASF-1 and ALF-1. These results suggest that the auxin responsiveness of the auxin-inducible genes parA, parB and parC are regulated by different mechanisms, even though the cis-acting elements look similar. PMID- 9729897 TI - Isolation of a novel NAD(P)H-quinone oxidoreductase from the cyanobacterium Synechocystis PCC6803. AB - A novel NAD(P)H-quinone oxidoreductase (NQR) was isolated from the cyanobacterium Synechocystis PCC6803 by ion-exchange, affinity and gel-filtration chromatographies. Isolated NQR was found to be a drgA gene product that was a homodimer composed of 23-kDa subunits. It showed NAD(P)H-plastoquinone oxidoreductase activity with Km values for NADPH and NADH of 12 and 48 microM, respectively. The activity was inhibited by thiolmodifying reagents, but not by rotenone, amobarbital, salicylhydroxamic acid, dicumarol, flavone, or diphenyleneiodonium chloride. Therefore, the Cys-147 residue is probably involved in the catalytic reaction. The amino acid sequence of the purified NQR had some homology with those of NADH oxidase, NAD(P)H-flavin oxidoreductase, and nitroreductase but did not contain either an adenine-binding motif or a phosphate binding motif, thus, it is a new type of NQR. PMID- 9729898 TI - Promoters of the phycocyanin gene clusters of the cyanobacterium Synechococcus sp. strain PCC 7942. AB - The cyanobacterium Synechococcus sp. strain PCC 7942 has duplicated phycocyanin subunit gene clusters cpcB1A1 and cpcB2A2, which are identical to each other and to those of Synechococcus sp. strain PCC 6301 (Anacystis nidulans). Nucleotide sequences of the 428 and 286 bases of the 5' non-coding regions of the cpcB1A1 and cpcB2A2 clusters, respectively, of strain PCC 7942 were identical to those of strain PCC 6301. As in strain PCC 6301, cpcB1A1 yielded two major transcripts of 1.4 and 1.3 kb and cpcB2A2 yielded a single transcript of 1.3 kb in strain PCC 7942. Thus, the structure and expression of cpcBA gene clusters in the two strains are essentially the same. Using bacterial luciferase encoded by luxAB as a reporter, cpcB1A1 was shown to have two promoters corresponding to the two major transcripts. Luminescence from the Synechococcus reporter strains carrying the fusions of the cpcBA promoters to luxAB showed circadian oscillation. Similar to the promoter of psbA1 encoding the D1 protein of PSII, the two cpcB1A1 promoters and the cpcB2A2 promoter showed the peak of activity at the end of the subjective day and the trough at the end of the subjective night. PMID- 9729900 TI - Factors affecting UV-B-induced changes in Arabidopsis thaliana L. gene expression: the role of development, protective pigments and the chloroplast signal. AB - Gene expression is known to change in response to UV-B radiation. In this paper, we have investigated three factors in Arabidopsis leaves that are likely to influence these changes: development, protective pigments and the 'chloroplast signal'. During late leaf development the major change in pigment composition, after exposure to UV-B radiation, is an increase in UV-absorbing pigments. Chl and Chl a/b ratio do not change substantially. Similarly Chl fluorescence is not altered. In contrast, RNA transcripts for photosynthetic proteins are reduced more in older leaves than in young leaves. To determine the role of flavonoids in UV-B protection, plants of Arabidopsis mutant tt-5, which have reduced flavonoids and sinapic esters, were exposed to UV-B and RNA transcript levels determined. The tt-mutants were more sensitive to UV-B radiation than wild-type. To examine the role of the chloroplast signal in regulating UV-B-induced changes in gene expression, Arabidopsis gun mutants (genome uncoupled) have been used. The results show that UV-B-induced down-regulation still takes place in gun mutants and strongly suggests that the chloroplast signal is not required. Overall, this study clearly demonstrates that UV-B-induced changes in gene expression are influenced by both developmental and cellular factors but not chloroplastic factors. PMID- 9729901 TI - Increase in the amount of celA1 protein in tobacco BY-2 cells by a cellulose biosynthesis inhibitor, 2,6-dichlorobenzonitrile. AB - The biochemical analysis of cellulose biosynthesis by plants has been a difficult problem due to the lack of a reliable assay procedure for cellulose synthase activity. Recently, the celA1 gene was cloned from cotton fiber, and this gene was identified from the rsw1 mutant of Arabidopsis as a catalytic subunit of cellulose synthase (Arioli et al. 1998). The cloning of these genes enables us to obtain specific antibodies against cellulose synthase. A highly specific antibody against celA1 protein was prepared and used to detect the protein from microsomal fraction of tobacco BY-2 cells. The quantity of celA1 protein in microsomal fraction of normal BY-2 cells was under the detection limit, although they contained a large quantity of cellulose. In contrast, cells habituated to 1 microM DCB (a specific inhibitor of cellulose biosynthesis) produced 1/10 of cellulose content of the normal cells, but had much more celA1 protein than the normal cells. The amount of polysaccharides in the EDTA-soluble fraction was relatively increased in habituated cells. The results suggest that celA1 protein is stabilized upon DCB binding and that the crystallization of cellulose microfibrils is inhibited simultaneously. PMID- 9729902 TI - Detection of the leghemoglobin gene on two chromosomes of Phaseolus vulgaris by in situ PCR linked-fluorescent in situ hybridization (FISH). AB - The leghemoglobin (Lb) gene on the metaphase chromosomes of Phaseolus vulgaris was amplified by in situ PCR. The amplified Lb gene could be detected on two chromosomes by fluorescent in situ hybridization (FISH) using the short Lb gene probe. PMID- 9729903 TI - Basing categorization on individuals and events. AB - Exemplar, prototype, and connectionist models typically assume that events constitute the basic unit of learning and representation in categorization. In these models, each learning events updates a statistical representation of a category independently of other learning events. An implication is that events involving the same individual affect learning independently and are not integrated into a single structure that represents the individual in an internal model of the world. A series of experiments demonstrates that human subjects track individuals across events, establish representations of them, and use these representations in categorization. These findings are consistent with "representationalism," the view that an internal model of the world constitutes a physical level of representation in the brain, and that the brain does not simply capture the statistical properties of events in an undifferentiated dynamical system. Although categorization is an inherently statistical process that produces generalization, pattern completion, frequency effects, and adaptive learning, it is also an inherently representational process that establishes an internal model of the world. As a result, representational structures evolve in memory to track the histories of individuals, accumulate information about them, and simulate them in events. PMID- 9729904 TI - Developmental differences in rule learning: a microgenetic analysis. AB - Trial-by-trial strategy assessments and a microgenetic design were used to examine 4- and 5-year-olds' learning of rules for solving balance scale problems. The design allowed us to examine simultaneously the contribution to rule learning of distal variables (qualities and knowledge with which children enter the learning situation) and proximal variables (processes that they execute during learning). Developmental differences in learning arose through two distal variables that were correlated with age--initial rule use and initial encoding helping older children to execute several proximal processes--noticing the potential explanatory role of a key variable, formulating a more advanced rule, and generalizing and maintaining the rule. Joint consideration of distal and proximal influences seems likely to be generally useful for understanding learning and development. PMID- 9729905 TI - [Use of collagen with gentamycin in abdominal and thoracic surgery]. AB - Based on an analysis of a group of 16 patients operated at the Surgical Clinic of the Third Medical Faculty, Charles University Prague the indication of collagen with gentamycin was established. Collagen with gentamycin is used locally for prophylaxis and treatment of intraabdominal and intrathoracic infections. In a group of patients it was used for prophylaxis of postoperative infections in elective abdominal operations such as plastic operations of the abdominal wall, anastomoses in the aboral part of the GIT, in operations of fistulae as part of treatment of intraabdominal abscesses and advanced cholecystitis and appendicitis. In thoracic surgery for prophylaxis of postoperative infections in plastic operations of the thoracic wall and also after surgery on account of inflammatory complications (lung abscess, bronchopleural fistula). PMID- 9729907 TI - [Complications of splenectomy]. AB - The authors describe the problematic of postsplenectomy complications. The main group of complications came in the group of haematological indication to splenectomy. The high risk of complications present the haematological malignancy. The most serious illness complications came only in this group. PMID- 9729906 TI - [Splenectomy--an analysis of problems]. AB - The authors discuss problematic of splenectomy on the historical view and also on the problematic indication to splenectomy. They compare two different periods in the small lapse of time. In the last period was the mowe in the indication spectrum to the splenectomy in the continuity with the application of new hematological medicaments in the practice. The number of "traumatological" splenectomy is constant. PMID- 9729908 TI - [An unusual complication after laparoscopic cholecystectomy]. AB - The authors describe an interesting complication after laparoscopic cholecystectomy. During a complicated operation perforation of a gallbladder, severely altered by inflammation, occurred and part of the concrements escaped into the peritoneal cavity. Several concrements were left in the abdominal cavity. After several weeks of a complicated postoperative development the concrements penetrated into the subcutaneous layer in the lumbar region and finally they were eliminated spontaneously. In the discussion the authors deal with complications of laparoscopic cholecystectomy. PMID- 9729910 TI - [Cholestasis--an indication for surgical treatment]. AB - Disorders of common bile duct function are among adults relatively frequent. Formal treatment schemes are gradually transformed, and the interdisciplinary co operation becomes the basic principle. The goal is to recognise the type of the mechanic jaundice, followed by elimination of the obstruction or to ensure biliary drainage without delay. The authors describe indication criteria used on Surgery Department at Bohunice Hospital in Brno. They present their results of treatment of the obstructive jaundice at a group of 576 patients followed during five year period. PMID- 9729909 TI - [Ileus caused by Recklinghausen's disease of the jejunum]. AB - Obstruction of the jejunum diagnosed during operation in a female patient with an ileous state. The patient had a history of loss of body weight, intermittent abdominal spastic pain in the umbilical region, fatigue, vomiting, histologically evaluated as a jejunal manifestation of Recklinghausen's disease. PMID- 9729911 TI - [When and how we perform surgery for inguinal hernia in children]. AB - In the course of 15 years in the authors department 4,694 operations of inguinal hernias and hydrocele were performed in children at the age of 0-15 years. Most frequently children aged 2-5 years were involved (45%). There were 4.5 times more boys than girls and the lateral distribution was also consistent with data in the literature. A relapse of hernia occurred in 33 children (i.e. 0.7%) operated in the authors department and in 19 children originally operated elsewhere. The total percentage of relapses in the group is 1.1%. In 25 children factors were identified which predispose for relapses. Relapses were most frequent in children operated before the age of one year (52%). The authors recommend therefore that operations of these children should be entrusted to a paediatric surgeon. PMID- 9729912 TI - [Dermatolipectomy of the abdominal wall]. AB - The authors describe clinical cases of dermatolipectmy of the abdominal wall performed during the last 15 years: a group of six female patients with excess skin after rapid and excessive reduction of adipose tissue, a second group of 42 patients with cumulation of subcutaneous fat in the region of the hypogastrium with formation of massive skinfolds, the so-called clinical picture of "venter pendulus". The third group of 122 patients with excessive adipose tissue and skin combined with diastasis of the rectus abdominis muscles or even prolapse of the abdominal organs. Transverse puboinguinal dermatolipectomy was performed. In 56 patients, high transverse resection is not used by the authors. Vertical lipectomy in the median line was performed in three patients and combined transverse-vertical lipectomy in 104 patients. From the total number of 170, operations satisfactory from the aspect of the surgeon and in particular the patient, were achieved in 122 (71.7%) cases, good results in 43 (25.4%) and satisfactory results in 5 (2.9%). PMID- 9729913 TI - [The effect of the primary operation on reoperations on the thyroid gland]. AB - The authors analyze the possibility to reduce the number of reoperations of the thyroid gland, based on a group of thyroid operations performed at the Surgical Clinic of the Second Medical Faculty Charles University and Faculty Hospital Motol during the 22-year period from 1975-1997. 507 reoperations from a total number of 4501 thyroid operations amount to 11.3%. In the authors opinion correct indication of the operation is most important as well as the correct extent and technique of the primary operation. The achieved results are except for unilateral temporary lesions of the NLR comparable with primary operations. PMID- 9729914 TI - [Aspergilloma]. AB - The authors present the case-history of a patient operated on account of a pulmonary aspergilloma, incl. analysis of postoperative infectious complications. The course is confronted with data in the literature. Based on the professional literature on the subject the authors discuss the contemporary approach to surgical treatment of this disease. PMID- 9729915 TI - [Malignant melanoma of soft parts (clear cell sarcoma)--a rare case of multiorgan localization]. AB - Malignant melanoma of soft parts (MMSP) is a rare tumor originally described by Enzinger in 1965 as clear cell sarcoma of tendons and aponeuroses because of its affinity to tenosynovial structures. Tumors are found predominantly at the extremities. First visceral case was described in 1993 in the duodenum. We describe the case of 64-years old man with malignant melanoma of soft parts in the stomach, in the pancreas, in the mesocolon, in the left thigh and in the left axilla. This patient was successfully treated surgically by the resection of the stomach, resection of the pancreatic head, extirpation of the tumor from mesocolon, from the left thigh and from the left axilla. In all these localisations the tumor was histologically and imunohistochemically proved to be MMSP (positivity: s-100 protein, vimentin, HMB-45 and negativity CK, EMA, desmin, actin). This multivisceral occurrence is extremely rare and according to the review of literature this is probably the first published case of MMSP in the stomach and in the pancreas. PMID- 9729916 TI - The quartz hazard: a variable entity. AB - An IARC Working Group recently classified crystalline silica (quartz) into IARC's Group 1, i.e. a carcinogen. This classification is based on evidence of carcinogenicity in experimental animals and in humans. However, the evaluation stated that in making the overall evaluation, the Working Group noted that carcinogenicity to humans was not detected in all industrial circumstances studied and that carcinogenicity may be dependent on inherent characteristics of the crystalline silica or on external factors affecting its biological activity. The present review seeks to put the apparently conflicting findings of cancer incidence in quartz-exposed industries into a unifying thesis, based on mechanistic studies. These mechanistic studies have enabled the events leading from deposition of quartz to silicosis and cancer to be partially elucidated and have demonstrated that the biological effects of quartz can be understood in terms of surface reactivity. We particularly emphasise the ability of quartz to generate free radicals and cause oxidative stress and the fact that this could be modified by a range of substances that affect the quartz surface; some of these modifying substances could originate from other minerals. We therefore propose that the hazard posed by quartz is not a constant entity, but one that may vary dramatically depending on the origin of the silica sample or its contact with other chemicals/minerals within its complex constitution. The mechanistic data described here could assist in the interpretation of epidemiological studies and pose further hypotheses that could be tested in order to help resolve the quartz carcinogenesis anomaly. The data suggest that quartz cannot be death with as a single hazard entity, as is the case with most other chemicals. PMID- 9729917 TI - Workplace exposure to rosin-based solder flux fume during hand soldering. AB - The patterns and extent of exposure to rosin based solder flux fume have been investigated in two surveys and a number of individual site visits carried out by the UK Health and Safety Executive (HSE). Determination of solder fume was by measurement of airborne resin acid particulate. Both static and personal sampling was carried out over time periods ranging from 15 minutes to several hours. Resin acid concentrations were found to vary from less than 1 microgram m-3 to 2289 micrograms m-3. The effects of various types of local exhaust ventilation on resin acid concentrations have been observed. On-tool tip extraction systems were generally found to be the best control measure available; however good design, positioning and system maintenance is essential for efficient capture of the fume. The resin acid concentrations detected at these twenty-six sites suggest that the proposed British long and short term occupational exposure limits are realistically attainable targets, particularly where good working practices and/or effective fume control measures are in place. PMID- 9729918 TI - A survey of wind speeds in indoor workplaces. AB - The applicability of the inhalable convention for sampling aerosols relies on its being a valid model for typical sampling environments. The current convention is based on measurements carried out in external wind speeds between 1 and 4 m.s-1. However these measurements show a degree of wind speed dependence, and it is uncertain at present how valid the convention is for describing human aspiration efficiency outside these wind speed limits. Following concerns that wind speeds in many indoor workplaces may be significantly below this range, measurements have been made in 55 work areas covering a wide range of workplaces. Measurements have concentrated on 'background' wind speeds where the influence of specific air movement sources is minimised. The pooled wind speed measurements show a highly skewed distribution with an arithmetic mean of approximately 0.3 m.s-1. Approximately 85% of all individual measurements were below this mean value. No obvious correlation was found between wind speed distribution parameters and industry type, room size or ventilation type. A limited number of comparisons were made between static anemometers and devices mounted on workers. It was found that modal wind speeds experience by workers were typically 0.05 m.s-1 higher than those measured using a static anemometer. These measurements agreed well with previously published data for similar workplaces as well as houses. PMID- 9729919 TI - Standards for environmental, non-threshold, carcinogens: a comparison of the approaches used for radiation and for chemicals. AB - Environmental standards for ionising radiation and for chemical carcinogens have been developed independently of each other. Radiation standards have been derived by deciding upon what is an acceptable risk, and then finding the corresponding dose from the exposure/risk relationship-quantitative risk assessment (QRA). The extent and the quality of the exposure/risk data for radiation, and the authority of the recommendations of the International Commission on Radiological Protection (ICRP), have resulted in universally accepted guidance and standards. This is not the case for chemical, non-threshold carcinogens. Their carcinogenicity ranges from doubtful to well-established, the exposure/response data are generally of poor quality, and there is no authoritative international body analogous to the ICRP. For some of these carcinogens, some organisations have used QRA to derive environmental standards. Others consider the data inadequate for such an approach and have used more pragmatic methods. The problems associated with the various approaches used and the prospects of an integrated approach for both radiation and chemical carcinogens are discussed. PMID- 9729920 TI - Assessment of triethylamine and diethylamine emission from military gas mask canisters. AB - A study was undertaken to evaluate the suitability of four types of amine modified charcoal filter canisters (cartridge) for use with gas masks (respirators) by measuring emissions of triethylamine (TEA) and diethylamine (DEA). Sampling and analysis methods for TEA and DEA were validated and optimized to ensure accurate measurement at low levels. A total of 88 air samples were taken by the validated methods to measure concentrations of TEA and DEA emitted from gas mask canisters during simulated use in an environmental chamber. Samples were collected on a mannequin equipped with a breather pump and also on human volunteers. Very low levels of TEA and moderately low levels of DEA emissions were measured during the simulations. The C7 (KOH-TEA-BPL/ASC3T) gas mask canister giving the lowest emission has been selected for use by the Canadian Forces. The potential health hazard from TEA and DEA for soldiers using the gas masks, under normal conditions, based on estimated use of one to two days per month, for a maximum of 4 hours/day for a normal working lifetime, was considered to be minimal and acceptable. PMID- 9729921 TI - Magnetic fields on British trains. AB - People on trains can be exposed to static and alternating magnetic fields which are higher than background levels in most homes and many workplaces. Quantification of such exposure may be of interest for epidemiological purposes but it is also important to ensure that exposure guidelines are complied with. This article describes the types of electric trains and trams in use in the UK and the results of measurements of static and alternating magnetic flux density. Many of the data have been supplied by the operators of the systems described. The measurements summarised in this article are indicative of the magnitudes of magnetic field exposures to be encountered on British trains, but without concomitant frequency information, they are not sufficient to allow demonstration of compliance with exposure standards. PMID- 9729922 TI - Respirable fibres: surfactant coated fibres release more Fe3+ than native fibres at both pH 4.5 and 7.2. AB - Exposure to asbestos is associated with several lung diseases. The carcinogenic action of asbestos is not fully understood but oxidative stress is considered to play a role. Iron on the surface can lead to Fenton chemistry and the Haber Weiss reaction producing free radicals such as the hydroxyl radical, which is likely to be important. Little is known of the pathogenic action of man-made fibres. This study involved the use of a panel of man-made fibres, some of which were shown to be pathogenic and others shown to be non-pathogenic in recent animal studies. A short term assay measuring Fe3+ release from the fibres over a 20 hour time period, and also a longer study of 12 week, found that pathogenic and non pathogenic fibres could not be differentiated according to Fe3+ release only. Iron release from native fibres was compared with that from surfactant-coated fibres, and in all cases surfactant coated fibres released more Fe3+ inferring that in vivo lung lining fluid coats native fibres and therefore affects the fibre surface chemistry and hence reactivity. PMID- 9729923 TI - Quantitative risk assessments derived from occupational cancer epidemiology: a worked example. PMID- 9729924 TI - Perceptions of absence from work: People's Republic of China versus Canada. AB - Cross-cultural theory was marshaled to predict how views of absence from work would be similar and different in Canada and the People's Republic of China. Respondents (N = 1,209) from both cultures had self-serving perceptions of their own absence levels, seeing them as exemplary compared with those of their work group and occupational peers. The Chinese showed a stronger tendency to generate estimates that favored their work group. Both cultural groups underreported their own actual absence. Chinese managers and employees agreed on absence norms, whereas Canadian managers provided lower estimates than did employees. Canadians and Chinese ranked the legitimacy of reasons for absence and attendance fairly similarly, but ratings showed that Canadians were less likely to endorse domestic reasons for absence, whereas Chinese were less likely to endorse illness, stress, and depression. PMID- 9729925 TI - Perceived team and player efficacy in hockey. AB - This study was designed to examine both the pattern of team and player efficacy across a season of competition and the relationships among player efficacy, team efficacy, and team performance in collegiate ice hockey. The team and player efficacies of hockey players from 6 teams in a midwestern collegiate hockey league were assessed prior to 32 games. Official game statistics were factor analyzed to produce one useable performance measure, performance outcome. A consensus analysis demonstrated that players held homogeneous beliefs regarding their own and their teams' abilities to perform successfully. A meta-analysis of the regression equations for each team confirmed the homogeneity among teams and the predictive superiority of team efficacy in predicting team performance. Also, when team wins and losses were analyzed across the season, team efficacy significantly increased after a win and significantly decreased after a loss, but player efficacy was not affected. PMID- 9729926 TI - Predictors of work injuries among employed adolescents. AB - Predictors of work injuries were studied in a sample of employed adolescents. The 20 predictors comprise 5 general categories of risk factors: demographic, personality, employment, health, and substance use. Data were obtained from a sample of 319 individuals ages 16 to 19. Hierarchical regression analysis revealed that all 5 categories of risk factors were related to job injuries. The significant predictors of work injuries among adolescents were gender, negative affectivity, job tenure, exposure to physical hazards, excessive workloads, job boredom, poor physical health, and on-the-job substance use. PMID- 9729927 TI - Relief from job stressors and burnout: reserve service as a respite. AB - To reveal the ameliorative impact of being away from job stressors on burnout, we compared 81 men who were called for active reserve service with 81 matched controls in the same company who were not called during the same period. Each reservist and his control completed questionnaires shortly before the reservist left work for a stint of service and immediately on his return. Analysis of variance detected a significant decline in job stress and burnout among those who served and no change among the control participants. Among those who served, quality of reserve service and degree of psychological detachment from work interacted in moderating the respite effects; the greater the detachment, the stronger the effect positive reserve service experience had in relieving reservists from stress and burnout. Reserve service is discussed as a special case of stress-relieving get-away from work that may be experienced as an ameliorative respite akin to vacation. PMID- 9729928 TI - Testing a theoretical model for examining the relationship between family adjustment and expatriates' work adjustment. AB - Based on theoretical perspectives from the work/family literature, this study tested a model for examining expatriate families' adjustment while on global assignments as an antecedent to expatriates' adjustment to working in a host country. Data were collected from 110 families that had been relocated for global assignments. Longitudinal data, assessing family characteristics before the assignment and cross-cultural adjustment approximately 6 months into the assignment, were coded. This study found that family characteristics (family support, family communication, family adaptability) were related to expatriates' adjustment to working in the host country. As hypothesized, the families' cross cultural adjustment mediated the effect of family characteristics on expatriates' host-country work adjustment. PMID- 9729929 TI - Perceptions of politics: does measuring different foci matter? AB - Recent research on perceptions of politics in organizations and other organizational phenomena (e.g., commitment) has suggested the use of a multiple foci approach to understand important politics-outcome relationships. This study confirms separate measures of perceptions of politics at the organizational and work-group levels and demonstrates differential effects in the prediction of various outcomes. After controlling for the effects of the relationship with one's supervisor (leader-member exchange), perceptions of politics existing at the organizational level predicted turnover intentions, whereas citizenship behavior was predicted by perceptions of politics at the group level. Both foci of politics significantly predicted organizational commitment. PMID- 9729930 TI - Cost-effectiveness of an intensive pressure ulcer prevention protocol in long term care. AB - Conducted in a 77-bed long-term-care facility, this study compared the costs of implementing an intensive pressure ulcer prevention protocol plus the calculated costs of treatment before and after implementing the protocol. A total of 69 patients comprised the preprotocol sample; 16 of them developed 26 ulcers. The postprotocol sample consisted of 63 patient, 3 of whom developed 5 ulcers. The 6 month pressure ulcer incidence was 23% in the preprotocol sample and 5% in the postprotocol sample. Mean cost for prevention and treatment of pressure ulcers was $113 +/- $345 per subject for the preprotocol sample and $100 +/- $157 per subject for the postprotocol sample (t = 0.27, df = 130, p = .79). Mean time to ulcer development was 146 +/- 61 days for the preprotocol subjects and 158 +/- 53 days for the postprotocol subjects (log rank = 8.63, p = .003 Implementation of a protocol that emphasized pressure ulcer prevention significantly reduced the incidence of pressure ulcers and cost per day of ulcer-free life. PMID- 9729931 TI - Underutilization of pressure ulcer risk assessment in hip fracture patients. AB - Patients with hip fractures are at high risk for pressure ulcer development, yet few recent studies have addressed risk assessment in this population. In this retrospective medical record survey, records of a random sample of California Medicare beneficiaries with hip fractures admitted to acute care hospitals in 1995 were selected for abstraction. Medical record abstraction (n = 545) revealed that risk assessment was performed on only 44.2% (241/545) of patients. Among these, 64.3% were identified as at risk for pressure ulcer development. Accuracy of the risk assessment was evaluated independently and compared with those performed by the nursing staff; there was agreement in only 40% of cases. Ulcers developed in 19.1% of the sample, and pressure ulcer development was not related to risk status at hospital admission. Patients with longer hospital stays developed more ulcers than those discharged earlier. This study shows that risk assessment is underutilized, inaccurate, and unrelated to pressure ulcer development in hip fracture patients. PMID- 9729932 TI - Use of a glycerin-based gel sheeting in scar management. AB - Management of hypertrophic and keloid scars can be difficult. This article presents an initial report on use of a glycerin-based gel sheeting to manage or prevent these scars. The results were comparable to those achieved with silicone sheeting, a typical treatment. Clinically, the glycerin-based sheeting was well tolerated, appeared to be effective, and was less expensive than the silicone sheeting. A prospective study will be done to confirm these findings. PMID- 9729933 TI - Where's the evidence for HCFA's policy? PMID- 9729934 TI - The importance of accurate information. PMID- 9729935 TI - The case for evidence-based practice standards. PMID- 9729936 TI - The growing influence of wound care teams. PMID- 9729937 TI - The team approach in diabetic foot management. AB - A significant reduction in the incidence of ulceration, infection, and lower extremity amputation can be realized through the institution of an organized foot care service in community and major academic medical centers. A multidisciplinary team approach has proven to be the most effective means of providing treatment and preventing foot lesions in the diabetic patient. Aside from prevention and early intervention, education is an essential component in overall patient management. In the scheme presented in this article, outpatient management is optimized through the services of numerous specialists dedicated to limb preservation. Risk factors must be evaluated, risk status determined, and preventive measures taken to preserve an intact foot. Ulcers must be thoroughly evaluated and appropriately treated through established protocols utilizing all members of the team. When acute problems present, they are more efficiently managed and coordinated by this approach, thereby reducing lengths of hospital stay, morbidity, and loss of limbs. PMID- 9729938 TI - The multidisciplinary in-hospital wound care team: two models. AB - The cost of community- and hospital-acquired pressure ulcers is particularly high in terms of both patient morbidity and economics. Multidisciplinary wound care teams were developed independently at two different hospitals to deal with the needs of patients with pressure ulcers and to control costs. Although the goals of the teams at both institutions were similar, the strategies for achieving the goals were different because they were adapted to the needs of the particular institution. As a result, care and prevention of pressure ulcers have improved at both hospitals. PMID- 9729939 TI - Obstacles and opportunities for the multidisciplinary wound care team. A report for the clinical symposium on wound management. AB - On the last day of the 12th Annual Clinical Symposium on Wound Management, a panel of clinicians from various disciplines, and with diverse experience in wound management, discussed the challenges and rewards of being part of a multidisciplinary team caring for patients with wounds. Panelists included Sharon Baranoski, MSN, RN, CETN; C. Andrew Salzberg, MD; Marlys J. Staley, MS, PT; and David R. Thomas, MD, FACP. Elizabeth A. Ayello, PhD, RN, CS, CETN, was the moderator. An excerpt from this session is published here. PMID- 9729940 TI - Pressure ulcer risk assessment scales--the missing link. PMID- 9729941 TI - Looking to the future of diabetic wound care. PMID- 9729943 TI - Defining loss of protective sensation in the diabetic foot. AB - Although the importance of distal symmetric sensorimotor polyneuropathy as a risk factor for diabetic foot ulceration has been known for decades, attempts to identify the elusive point that defines loss of protective sensation have been futile. This manuscript will review the epidemiology and pathogenesis of peripheral neuropathy, discuss commonly used screening tools, and review recently reported data that more sharply define loss of protective sensation on the long spectrum of neuropathy. PMID- 9729942 TI - Evaluation of a collagen-alginate wound dressing in the management of diabetic foot ulcers. AB - Efficacy and safety of a collagen-alginate topical wound dressing (FIBRACOL Collagen-Alginate Wound Dressing) in the treatment of diabetic foot ulcers was compared with that of regular gauze moistened with normal saline. Seventy-five patients with foot ulcers were assigned randomly in a 2:1 ratio to the collagen alginate test dressing or the gauze dressing. At the end of the study, the mean percent reduction of the wound area was 80.6% +/- 6% in the collagen-alginate dressing group and 61.1% +/- 26% in the gauze dressing group (p = .4692). Thirty nine (78%) patients treated with the collagen-alginate dressing achieved > or = 75% wound area reduction, compared with 15 (60%) of gauze-treated patients. Complete healing was achieved in 24 (48%) of the collagen-alginate dressing group and 9 (36%) of the gauze dressing group. Wound size, when averaged over the 8 week period and with the duration of the ulcer taken into account, was reduced significantly in the collagen-alginate dressing group, as compared with the gauze dressing group (df = 1, p = .0049). It is concluded that the collagen-alginate test dressing is as or more effective and safe as the currently used treatment. PMID- 9729944 TI - Use of layered compression bandages in diabetic patients. Experience in patients with lower leg ulceration, peripheral edema, and features of venous and arterial disease. AB - Layered compression therapy for venous leg ulcers and ulcers associated with chronic leg edema has been shown to be an effective treatment in patients with adequate arterial circulation. However, no study has looked specifically at compression therapy in the diabetic population. This clinical case review examines outcomes in two groups of diabetic patients with edema and either venous ulcers or preulcerative conditions. Patients in Group 1 had clinically adequate arterial circulation and were treated with a four-layer compression bandage system. The highly elastic third layer was eliminated in the Group 2 patients, who had compromised peripheral arterial circulation. Healing occurred in 81% of patients in Group 1 and 67% of patients in Group 2. There was no acute progression of lower limb ischemia. Layered compression therapy was an effective and safe treatment in this diabetic population with adequate arterial circulation. Reduced compression also can be helpful in some patients with arterial compromise. PMID- 9729945 TI - Clinical research opportunities in long-term care. PMID- 9729947 TI - Pressure and blood flow linkages and impacts on pressure ulcer development. PMID- 9729946 TI - The impact of pressure ulcers on health care costs and mortality. PMID- 9729949 TI - Risk and assessment of pressure ulcer development in surgical patients. PMID- 9729948 TI - Pressure ulcer risk factors in the operating room. PMID- 9729950 TI - Intraoperatively acquired pressure ulcer prevalence: a national study. PMID- 9729951 TI - Prospective study of the incidence of OR-induced pressure ulcers in elderly patients undergoing lengthy surgical procedures. PMID- 9729952 TI - Study results: prediction and prevention of pressure ulcers in surgical patients. PMID- 9729953 TI - OR-acquired pressure ulcers in vascular surgery patients. PMID- 9729954 TI - A pressure ulcer toolbox for facilitating hospital-wide quality. PMID- 9729955 TI - Preliminary results of a randomized, controlled study of a pressure ulcer prevention system. PMID- 9729957 TI - Is there an answer to the medical review policy nightmare? PMID- 9729956 TI - A comparative, randomized, controlled study to determine safety and efficacy of preventive pressure ulcer systems: preliminary analysis. PMID- 9729958 TI - Another fine mess... PMID- 9729959 TI - Two hospice quality of life surveys: a comparison. AB - This study's objective tested the utility of two quality of life (QOL) forms in a hospice setting. The compared forms were the McGill Quality of Life Questionnaire (MQOL) and the Hospice Quality of Life Index-Revised (HQLI). Using a crossover design, hospice nurses first administered one survey to eligible patients and then, in the study's second phase, administered the other survey to newly enrolled eligible patients. Nurses were interviewed regarding each form and possible changes in patient care that were made due to the assessment. Hospice care plans were reviewed looking for specific changes as a result of the surveys. The results showed that the QOL assessments were useful for the nurses in planning the care of the hospice patients and that the MQOL was preferred by the nurses over the HQLI. PMID- 9729960 TI - Occupational stress in hospice care: causes and coping strategies. AB - Thirty-three nurses from three hospice services in a large Midwestern city participated in this study, which investigated responses to difficult or demanding work-related situations. Three tools (the Self Inventory of Situational Responses-TC questionnaire, the Spielberger State Anxiety Inventory questionnaire and rank-ordered listing of likely causes of difficult or demanding situations) were used to collect data. Findings indicated that anxiety is an issue for hospice workers but that difficult or demanding situations were viewed as challenges rather than threats. Management of intractable symptoms and communication issues were of primary concern. Administrative concerns were identified as the third-ranking source of difficult situations. Issues related to death and dying were of notably less concern. Strategies for staff support also are identified. PMID- 9729962 TI - Personal, protective and cost-effective infection control: an overview for hospice facilities. AB - This article has presented some tips for developing an exposure control/infection control program that is practical, protective and cost-effective. OSHA state that "employers may create the most protective and cost-effective programs possible." We do not jeopardize employee safety by using science to create a cost-effective program. In order for staff to understand this issue, proper education and training must be provided. Often times agencies look for the "quick fix", and that may come back to haunt you. Send time conducting a proper assessment of risk and needs and develop a program that is a "win-win" for both staff and management. PMID- 9729961 TI - Review of Medicare's proposed hospice eligibility criteria for select noncancer patients. AB - Recent events have challenged our health system to increase access to and provide high quality care for patients near the end of life. Simultaneously, Medicare is developing review policies to determine eligibility for hospice patients with select noncancer diagnoses. The purpose of this study was to determine whether the proposed policies met one of their chief goals: accurate identification of patients with a less-than-six-months prognosis. Only 35 percent of 104 patients who died within six months of admission to the hospice used for this study, LifePath Hospice, met the Medicare proposed criteria for hospice eligibility. The median and mean survival time of the sample was 14 and 30 days respectively. Based on this review, it is recommended that Medicare alter their proposed review policies and not limit access to hospice eligible patients who desire and are in need of such services. PMID- 9729964 TI - Bibliography for music therapy in palliative care, 1963-1997. PMID- 9729963 TI - Spiritual terrorism. AB - Spiritual abuse is the act of making people believe--whether by stating or merely implying--that they are going to be punished in this life and/or tormented in hell-fire forever for failure to live a good enough life to earn admission to heaven. Spiritual terrorism is the most extreme form of spiritual abuse, which in itself is a serious mental health problem. The 12 steps of Alcoholics Anonymous can be a useful therapeutic modality for countering this problem, provided step two and three are reworded to facilitate cognitive restructuring, therefore enabling victims to develop a positive conception of God. Biblical symbolism can be cognitively helpful if interpreted metaphorically rather than literally. Thus, victims will be able to trust God to restore them to sanity and empower them to be survivors who experience peace of mind, joy of living and freedom from fear. PMID- 9729965 TI - Facing the challenge of the hospice and palliative care marriage. PMID- 9729966 TI - Professional education in hospice and palliative care. PMID- 9729967 TI - The homeless terminally ill and hospice & palliative care. PMID- 9729968 TI - International palliative care 1998. PMID- 9729969 TI - Clamping down on nursing home contracts. PMID- 9729970 TI - Questioning degrees of 'low-tech'. PMID- 9729973 TI - Economics of unrelieved cancer pain. PMID- 9729972 TI - A prospective study on the dying process in terminally ill cancer patients. AB - To determine the physical and medical change in the dying process, a prospective study was performed on 100 terminally ill cancer patients. The mean (median) time from the onset of death rattle, respiration with mandibular movement (RMM), cyanosis on extremities, and pulselessness on the radial artery to death was 57 (23) hours, 7.6 (2.5) hours, 5.1 (1.0) hours, and 2.6 (1.0) hours respectively. Death rattle preceded the other three conditions in 74 percent of the subjects, while RMM preceded cyanosis and pulselessness in 63 percent. The ratio of awake drowsy-comatose patients was 56-44-0 percent one week before death, 26-62-12 percent in the last 24 hours, and 8-42-50 percent in the final six hours. The number of opioid users and average dose increased significantly as death approached, from 42 percent and 49 mg/day (parental morphine equivalent) four weeks before death to 87 percent and 139 mg/day in the final 48 hours. The frequency of extra dosage also increased significantly, from 32 percent (opioid) and 40 percent (non-opioid) one week before death to 68 percent and 66 percent in the last 48 hours, respectively. The change of physical signs and medical intervention when death is impending has a common pathway in spite of large individual variations; thus, understanding this nature can help clinicians to offer better palliative care to terminal cancer patients. PMID- 9729971 TI - Implementation and evaluation of a quality improvement process to improve pain management in a hospice setting. AB - The purpose of this article is to describe the implementation and evaluation of a quality improvement process to improve pain management in a hospice setting. A retrospective chart audit of 702 patient visits pre- and 536 patient visits post implementation of quality improvement strategies measured five aspects of pain management: complaints of pain, severity of pain, changes in patient's pain medication regime required, patient and family teaching, and use of complementary therapies. Of these measures a significant change was found in the documentation of pain assessment, recognition of changes required in the medication regime, and patient and family teaching. PMID- 9729974 TI - Spiritual abuse. AB - Spiritual abuse is the act of making people believe--whether by stating or merely implying--that they are going to be punished in this life and/or tormented in hell-fire forever for failure to live life good enough to please God and thus earn admission to heaven. Spiritual terrorism is the most extreme form of spiritual abuse and may cause serious mental health problems. Those people who have not been spiritually terrorized have not necessarily been spared from spiritual abuse and therefore may still be in need of competent, spiritual counseling. Spiritual abuse, which may be active or passive, can best be conceptualized on a continuum from terroristic to zero abuse. Severity is determined by intensity, age of onset, duration, and individual reaction. The underlying issue in all forms of abuse is control. PMID- 9729976 TI - Patient education: adding to quality of care. PMID- 9729978 TI - Explorers and builders: musings on nursing research towards the year 2000. PMID- 9729977 TI - Conflict can be good. PMID- 9729975 TI - Management of intestinal obstruction in hospice care. AB - Bowel obstruction in the terminally ill is a relatively common complication. The classic therapy of i.v. hydration with nasogastric suction is contraindicated due to inefficacy and patient discomfort. Modern therapy consists of state-of-the-art pharmacologic treatments, as well as the judicious use of surgical interventions in selected cases. Many patients can be relatively symptom-free and can eat and drink as tolerated. PMID- 9729979 TI - University-based psychiatric nursing education: a promise for the future? AB - University-based nursing education was introduced in Victoria, Australia, to redress the deficiencies attributed to the system of hospital-based training. The narrow and restrictive focus, maintenance of the subservient position of nurses, the employee status of students, an inadequate relationship between theory and practice and failure to keep pace with changes in the role of the nurse were the main deficiencies identified. This paper refers to a qualitative research study which examines the differences between graduates of a university-based and hospital-based psychiatric nursing program during the year following graduation. The findings suggest that the tertiary-based course had the potential to significantly redress some inadequacies of the hospital-based course to produce different qualities within its graduates. PMID- 9729980 TI - Clinical teaching models: a review of the role of preceptor in the undergraduate nursing program. AB - The relatively recent transition of nursing education in Australia from a hospital-based apprenticeship, into the higher education sector has been marked by an acknowledgement that the clinical knowledge of undergraduate student nurses is at risk of becoming separated from their theoretical knowledge. The length of time which student nurses spend on clinical placements experiencing the complexities of the clinical world is diminishing, partly in response to economic constraints on universities and health care agencies. This contrasts with an expectation that students will be safe and competent practitioners immediately on graduation. The preceptor model is being increasingly used in the final semester of the undergraduate-nursing course to facilitate the development of critically thinking, reflective practitioners. This paper examines some of the assumptions that have been made about preceptorship in its adoption into the Australian nursing education context. It is concluded that further research into the experiences of registered nurses in the preceptor role may increase our understanding of the potential of preceptorship as a clinical teaching model which can meet the future needs of both nursing education and nursing service. PMID- 9729982 TI - CyberNurse. PMID- 9729981 TI - Collaborative research--can technology help? PMID- 9729983 TI - The clinical application of the nursing process in selected acute care settings: a professional mirage. AB - The nursing process is the problem solving framework purported to be used in a multiplicity of health care settings. Although its use is widespread in educational and clinical settings, some nurse clinicians display negative attitudes towards its use and state that it is incongruent with nursing practice. To date, there has been no study cited that has examined it use within clinical settings to determine the substance of these claims. Using grounded theory methodology, this study examined the clinical application of the nursing process in acute care hospital settings. Data were obtained from semi-structured interviews with predominantly nurse clinicians (n = 27), participant field observations of nurse clinicians, and in-depth audits of patient records. Textual data were managed using NUD-IST and analysed using constant comparative method. Data generation and analysis proceeded simultaneously using open coding, theoretical coding, and selective coding techniques until saturation was achieved. Nurses in this study experienced the basic social problem of being in a state of "Unknowing" that was linked to a number of factors, such as, the existence of a fragmented and inconsistent method of determining and communicating patient care and work conditions of immense change and uncertainty. The findings revealed several problems with the clinical application of the nursing process and illustrated that the espoused theory was unable to be clinically applied. PMID- 9729984 TI - The non-insulin-dependent diabetic: success and failure in compliance. AB - Failure to comply with medical regimes is endemic and is extremely costly both in economic and wellness terms. Adherence failure is explored in this study of 95 non-insulin-dependent-diabetics (NIDDM) subjects. Results indicate that the factors governing failure to adhere are not necessarily the obverse of psychosocial factors correlated with successful adherence to medical regimes. Success in complying to wellness behaviours was a function of positive attitudes and an acceptance of the challenges of the illness, the capacity to utilise family support, and not having negative self esteem. Failure to comply was a function of a syndrome of stress, chronic and transient mental distress. Common to both success and failure in complying were attitudes to health professionals, extent of knowledge about the disease and perceptions of the success or otherwise in overcoming inconvenience and barriers which might impede compliance. PMID- 9729985 TI - A piece of my mind. Hey! I'm a teacher too. PMID- 9729986 TI - Minority faculty and academic rank in medicine. AB - CONTEXT: Previous studies have found that fewer minority medical school faculty hold senior professorial ranks than do majority faculty and may not be promoted as rapidly. OBJECTIVE: To determine whether minority faculty were as likely as majority faculty to have attained senior rank (associate professor or full professor) after adjusting for other factors that typically influence promotion. DESIGN: A self-administered mailed survey of US medical school faculty using the Association of American Medical Colleges database. The sample was stratified by department, graduation cohort, and sex. PARTICIPANTS: A stratified random sample of 3013 full-time faculty at 24 representative US medical schools. All underrepresented minority faculty at these schools were sampled. MAIN OUTCOME MEASURE: Attainment of senior academic rank (associate professor or full professor). RESULTS: Of 3013 faculty surveyed, 1807 (60.0%) responded, including 1463 white (81.0%), 154 black (8.5%), 136 Asian (7.5%), and 54 Hispanic (3.0%). Overall, 980 faculty (54%) had attained senior academic rank, including 47 (30.5%) of 154 black faculty, 59 (43.4%) of 136 Asian faculty, 22 (40.8%) of 54 Hispanic faculty, and 852 (58.3%) of 1463 white faculty. White faculty had significantly more first-authored and total peer-reviewed publications than the other groups. After adjusting for the medical school, department, years as medical school faculty, number of peer-reviewed publications, receipt of research grant funding, proportion of time in clinical activities, sex, and tenure status, we found that the odds ratios of holding senior rank relative to white faculty were 0.33 (95% confidence interval [CI], 0.17-0.63) for black faculty, 0.36 (95% CI, 0.12-1.08) for Hispanic faculty, and 0.58 (95% CI, 0.30-1.12) for Asian faculty. CONCLUSIONS: Minority faculty were less likely than white faculty to hold senior academic rank. This finding was not explained by potential confounders such as years as a faculty member or measures of academic productivity. PMID- 9729987 TI - Effect of an intensive educational program for minority college students and recent graduates on the probability of acceptance to medical school. AB - CONTEXT: Increasing the number of minority physicians is a long-standing goal of professional associations and government. OBJECTIVE: To determine the effectiveness of an intensive summer educational program for minority college students and recent graduates on the probability of acceptance to medical school. DESIGN: Nonconcurrent prospective cohort study based on data from medical school applications, Medical College Admission Tests, and the Association of American Medical Colleges Student and Applicant Information Management System. SETTING: Eight US medical schools or consortia of medical schools. PARTICIPANTS: Underrepresented minority (black, Mexican American, mainland Puerto Rican, and American Indian) applicants to US allopathic medical schools in 1997 (N =3830), 1996 (N = 4654), and 1992 (N =3447). INTERVENTION: The Minority Medical Education Program (MMEP), a 6-week, residential summer educational program focused on training in the sciences and improvement of writing, verbal reasoning, studying, test taking, and presentation skills. MAIN OUTCOME MEASURE: Probability of acceptance to at least 1 medical school. RESULTS: In the 1997 medical school application cohort, 223 (49.3%) of 452 MMEP participants were accepted compared with 1406 (41.6%) of 3378 minority nonparticipants (P= .002). Positive and significant program effects were also found in the 1996 (P=.01) and 1992 (P=.005) cohorts and in multivariate analysis after adjusting for nonprogrammatic factors likely to influence acceptance (P<.001). Program effects were also observed in students who participated in the MMEP early in college as well as those who participated later and among those with relatively high as well as low grades and test scores. CONCLUSIONS: The MMEP enhanced the probability of medical school acceptance among its participants. Intensive summer education is a strategy that may help improve diversity in the physician workforce. PMID- 9729988 TI - Employment-seeking experiences of resident physicians completing training during 1996. AB - CONTEXT: Studies analyzing the physician workforce have concluded that the United States is verging on a physician oversupply, yet we lack persuasive evidence that this is resulting in physician underemployment and/or unemployment. OBJECTIVE: To determine the degree to which graduating residents have difficulty finding or are unable to find employment in their primary career choices. DESIGN: Two 1-page surveys sent separately to residents and to program directors to collect information on the employment status of residents who were completing a graduate medical education program at the end of the 1995-1996 academic year. SETTING: A total of 25 067 resident physicians scheduled in the spring of 1996 to complete a residency program accredited by the Accreditation Council on Graduate Medical Education, and 4569 program directors in 31 specialties and subspecialties. MAIN OUTCOME MEASURE: Both the graduates' employment status and the degree of difficulty they experienced securing a practice position, as reported by resident physicians and program directors. RESULTS: After 6 months of data collection, 12135 (48.4%) of 25 067 resident physicians responded to the survey. Of the respondents, 11 200 had completed their training, and 7628 (68.1%) were attempting to enter the workforce, 28.4% were seeking additional training, and 3.5% were fulfilling their military obligations. Of the 7628 resident physicians who sought employment, 67.3% obtained clinical practice positions in their specialties, 15.5% took academic positions, 5.0% found clinical positions in other specialties, 5.1% had other plans, and 7.1% did not yet have positions but were actively looking. In addition, 22.4% of resident physicians who found clinical positions reported significant difficulty finding them. The subgroup reporting greater difficulty finding clinical positions included international medical graduates (more than 40%),those completing programs in the Pacific or East North Central region, and those in several specialties. The 1996 graduating residents reported significantly higher rates of difficulty finding suitable employment than program directors reported for their graduates (22.4% vs 6.0%); however, the percentage of graduates reported by both groups as entering the workforce was the same (68.1%). Program directors reported an unemployment rate of only 1.2%, for their 1996 graduates, which was less than the rate reported by the resident physicians (7.1%). CONCLUSIONS: Resident physicians' direct reports of their employment-seeking experiences differ from what program directors report. Program directors accurately determined the number of residents pursuing further training; however, they did not have complete information about the employment difficulties experienced by their graduates. Based on graduates' reports, we conclude that employment difficulties are greatest among international medical graduates and vary by specialty and geographic region. PMID- 9729989 TI - Courses involving complementary and alternative medicine at US medical schools. AB - CONTEXT: With the public's increasing use of complementary and alternative medicine, medical schools must consider the challenge of educating physicians about these therapies. OBJECTIVES: To document the prevalence, scope, and diversity of medical school education in complementary and alternative therapy topics and to obtain information about the organizational and academic features of these courses. DESIGN: Mail survey and follow-up letter and telephone survey conducted in 1997-1998. PARTICIPANTS: Academic or curriculum deans and faculty at each of the 125 US medical schools. MAIN OUTCOME MEASURES: Courses taught at US medical schools and administrative and educational characteristics of these courses. RESULTS: Replies were received from 117 (94%) of the 125 US medical schools. Of schools that replied, 75 (64%) reported offering elective courses in complementary or alternative medicine or including these topics in required courses. Of the 123 courses reported, 84 (68%) were stand-alone electives, 38 (31%) were part of required courses, and one (1%) was part of an elective. Thirty eight courses (31%) were offered by departments of family practice and 14 (11%) by departments of medicine or internal medicine. Educational formats included lectures, practitioner lecture and/or demonstration, and patient presentations. Common topics included chiropractic, acupuncture, homeopathy, herbal therapies, and mind-body techniques. CONCLUSIONS: There is tremendous heterogeneity and diversity in content, format, and requirements among courses in complementary and alternative medicine at US medical schools. PMID- 9729990 TI - Current and projected workforce of nonphysician clinicians. AB - Nonphysician clinicians (NPCs) are becoming increasingly prominent as health care providers. This study examines 10 such disciplines: nurse practitioners (NPs), physician assistants (PAs), nurse-midwives, chiropractors, acupuncturists, naturopaths, optometrists, podiatrists, nurse anesthetists, and clinical nurse specialists. The aggregate number of NPCs graduating annually in these 10 disciplines doubled between 1992 and 1997, and a further increment of 20% is projected for 2001. Assuming that enrollments remain at the levels attained in 2001, NPC supply will grow from 228000 in 1995 to 384000 in 2005, and it will continue to expand at a similar rate thereafter. The greatest growth is projected among those NPCs who provide primary care services. Moreover, the greatest concentrations of both practicing NPCs and NPC training programs are in those states that already have the greatest abundance of physicians. On a per capita basis, the projected growth in NPC supply between 1995 and 2005 will be double that of physicians. Because of the existing training pipeline, it is probable that most of the growth projected for 2005 will occur. The further expansion of both NPC and physician supply thereafter warrants careful reconsideration. PMID- 9729991 TI - Roles of nonphysician clinicians as autonomous providers of patient care. AB - Studies were undertaken to assess the practice prerogatives of nonphysician clinicians (NPCs) in 10 disciplines that, collectively, are the major nonphysician contributors to the delivery of medical and surgical services. These disciplines include nurse practitioners, physician assistants, nurse-midwives, chiropractors, acupuncturists, naturopaths, optometrists, podiatrists, nurse anesthetists, and clinical nurse specialists. Marked differences were found in the practice prerogatives that states granted NPCs in the various disciplines. For most disciplines, the magnitude of their prerogatives correlated with the numbers of NPCs practicing in each state. At their maximal levels, state practice prerogatives authorized a high degree of autonomy and a broad range of authority to provide discrete levels of uncomplicated primary and specialty care. The recent growth in these prerogatives is fostering new opportunities for NPCs; however, it also is creating a pluralism that has the potential to further fragment the US health care system. It is time for regulatory integration and professional collaboration so that a health care workforce that includes a diversity of disciplines can be assured of providing a coherent set of patient care services in the future. PMID- 9729992 TI - Educational programs in US medical schools, 1997-1998. AB - To describe the current status of medical education programs in the United States, we used data from the 1997-1998 Liaison Committee on Medical Education Annual Medical School Questionnaire, which had a 100% response rate, and from other sources. There were 96733 full-time medical school faculty members, a 1.2% increase from 1996-1997. The 43020 applicants for the class entering in 1997 represents an 8.4% decrease from 1996. The number of 1997 applicants who were members of underrepresented minority groups decreased 11.1 % from 1996, and the number of entering underrepresented minority group students decreased 8.4%. More than half of medical schools reported that the number of inpatients available for medical student education had decreased in at least some of their clinical sites or in some disciplines during the past 2 years. Thirty-nine medical schools (31.2%) reported having more difficulty recruiting or retaining volunteer clinical faculty to participate in medical student teaching in 1997 than in 1995. PMID- 9729994 TI - Review of US medical school finances, 1996-1997. AB - Based on data from the Annual Medical School Questionnaire of the Liaison Committee on Medical Education, to which 100% of schools responded, the revenues that supported the programs and activities of the 125 accredited medical schools in the United States totaled $34897 million in 1996-1997. A large proportion (78.9%) of these revenues was derived from 3 sources: practice plans, grants and contracts, and hospital support. Both public and private medical schools, in aggregate, have continued to experience growth throughout the last decade but at a progressively slower rate, primarily because of a slowing in the growth of practice plan revenues. Federal revenues supporting research in public and private medical schools since 1992-1993 have grown at annualized, constant-dollar rates of 5.6% and 4%, respectively. Growth in state and local appropriations to public medical schools has tended to lag behind inflation. Growth in reported revenues from endowments that are used to support programs at private medical schools is on the rise. The aggregate numbers mask considerable variation among schools with regard to changes in financing. A small, but appreciable, number of schools have witnessed a constant-dollar decline in their total practice plan revenues since 1992-1993. The financial data reviewed in this report demonstrate the continued dependence of medical schools on faculty-generated sources of revenue and confirm the perception that medical schools, as a group, are experiencing constraints on the growth of their enterprises. PMID- 9729993 TI - Graduate medical education, 1997-1998. AB - In response to growing concerns that continued unlimited governmental funding of graduate medical education (GME) would lead to a physician surplus, Congress enacted provisions in the Balanced Budget Act (BBA) of 1997 to limit further growth, as well as to encourage reductions in GME. The measures incorporated in this section of the BBA reflect recommendations made by a number of major professional associations. The question now is how effective these efforts will be and whether they will produce unintended or deleterious consequences. We report the changes occurring in GME from 1993 to 1997, focusing on changes prior to and since the enactment of the BBA. The total number of residents in GME programs has remained relatively constant from 1993 to 1997. The number of residents entering GME programs without prior GME experience has also remained constant; however, over the same period, the number entering a new program with some prior GME experience has fallen by 5.8%. The number of international medical graduates in all GME programs has increased 12.4% during this same period, while the number of US allopathic medical school graduates has decreased 4.4%. As federal and state initiatives are introduced to change the number and distribution of GME positions, it is critical that the American Medical Association and other professional organizations monitor GME tracking data more systematically and accurately than ever before. PMID- 9729995 TI - Medical student financial assistance, 1996-1997. AB - Loans account for the major portion of financial aid available to medical students. In the academic year 1996-1997, 80.1% of all available financial aid came from loans, and medical students borrowed more than $1.11 billion. Of the 1997 medical school graduating class, 83.2% had educational debt, 46% of whom had mean educational debt levels higher than $75000. PMID- 9729996 TI - Time to shatter the glass ceiling for minority faculty. PMID- 9729997 TI - Creating an effective physician workforce marketplace. PMID- 9729999 TI - Online: the new Medical Student JAMA and a Web of possibilities. PMID- 9729998 TI - Physicians and nonphysician clinicians: complements or competitors? PMID- 9730000 TI - Prevalence of harassment and discrimination among 1996 medical school graduates: a survey of eight US schools. AB - CONTEXT: Harassing and discriminating behaviors on the part of instructors or supervisors are known to affect the quality of work performed by medical students, influence their career decisions, and have other undetermined long-term consequences. OBJECTIVE: To assess the prevalence and forms of harassment and discrimination experienced by 1996 medical school graduates. DESIGN: A self administered survey of harassment and discrimination mailed to graduating medical students. SETTING AND PARTICIPANTS: A total of 1001 graduating medical students at 8 US medical schools (4 public and 4 private), chosen from each of the 4 regions designated by the Association of American Medical Colleges for geographic categorization. OUTCOME MEASURE: The number of reported experiences of various forms of harassment and discrimination while attending medical school. RESULTS: Of 1001 surveys, 548 (55%) were returned. Overall, 46% of the students reported experiencing some form of harassment and 41% some form of discrimination from instructors or supervisors while attending medical school. Nonsexual verbal harassment was reported by 41%; sexual verbal harassment was reported by 10%. Discrimination based on gender was reported by 29% of students; discrimination based on race was reported by 12%. CONCLUSIONS: Harassment and discrimination of medical students by instructors and supervisors continue to occur frequently, and new approaches are needed to address these problems. PMID- 9730001 TI - The role of the television drama ER in medical student life: entertainment or socialization? PMID- 9730003 TI - JAMA patient page: medical education. PMID- 9730002 TI - An interview with Neal Baer, MD, the doctor behind ER. PMID- 9730005 TI - Steroidal muscle relaxants attenuate the contractile and phosphatidylinositol responses of rat trachea. AB - Interaction of the steroidal muscle relaxants, pancuronium, vecuronium and rocuronium with airway muscarinic receptors of rat trachea was investigated in vitro concerning the contractile and phosphatidylinositol (PI) responses. Pancuronium and vecuronium attenuated, while rocuronium did not affect carbachol (CCh)-induced contraction and CCh-induced IP1 accumulation. Pancuronium could inhibit completely CCh-induced contraction at a dose of 30 microM, while it could not inhibit completely CCh-induced IP1 accumulation at the same dose. These drugs attenuated KCl-induced contraction. These results suggest that pancuronium and vecuronium would attenuate airway smooth muscle contraction through the inhibition of muscarinic receptor-mediated PI response, and that the inhibition of voltage-operated Ca++ channel might be involved, in part, in the attenuation by pancuronium of the contraction. PMID- 9730004 TI - Granisetron, a 5-HT3 receptor antagonist, inhibited cisplatin-induced 5 hydroxytryptamine release in the isolated ileum of ferrets. AB - We investigated the influence of granisetron, a 5-HT3 receptor antagonist, on the increase in 5-hydroxytryptamine (5-HT) release induced by cisplatin from the isolated ileum of the ferret, a species known to vomit in response to cisplatin. 2-Methyl-5-HT, a selective 5-HT3 receptor agonist, increased the release of 5-HT from the ferret ileum in a concentration-dependent manner within the range of 10( 7) to 10(-6)M. The 5-HT release induced by 2-methyl-5-HT was significantly inhibited by a concomitant perfusion with granisetron (10(-6)M). Cisplatin also increased the 5-HT release from the ferret ileum within the range of 10(-8) to 10(-6)M, in a concentration-dependent manner. Granisetron (10(-6)M) also significantly inhibited the cisplatin-induced 5-HT release. Since the cisplatin induced 5-HT release was significantly inhibited by tetrodotoxin, the possible involvement of an interneuron pathway in the cisplatin-induced 5-HT release mechanism was suggested in the ileal tissue. It is likely that granisetron inhibited the cisplatin-induced 5-HT release from the gut EC cells by producing blockade of an EC cell 5-HT3 receptor. PMID- 9730006 TI - Salicylic acid-induced lipid peroxidation in rat liver microsomes. AB - Rat liver microsomes (1 mg protein/ml) were incubated at 37 degrees C with non steroidal anti-inflammatory drug, salicylic acid (5 mM) in the presence of 0.2 mM NADPH. The amounts of thiobarbituric acid reactive-substances (TBARS) were increased remarkably by the incubation for 30 min. TBARS formation was dependent on salicylic acid concentration. Salicylic acid-induced lipid peroxidation was not observed in the absence of NADPH. It was also inhibited by the presence of cytochrome P450 inhibitor, SKF-525A. The singlet oxygen's scavengers such as histidine suppressed salicylic acid-induced lipid peroxidation. These results suggested that salicylic acid-induced lipid peroxidation was coupled with the metabolism through cytochrome P450 in rat liver microsomes. The singlet oxygen may be involved in the reaction. PMID- 9730007 TI - Effect of pibutidine hydrochloride on nitric oxide production in rat gastric mucosa. AB - This study examined the effect of pibutidine hydrochloride, a novel histamine H2 receptor antagonist (IT-066), on nitric oxide (NO) production in gastric mucosa by measuring nitrite and nitrate contents. IT-066 (0.3-3 mg/kg, p.o.) and NO donors, sodium nitroprusside (0.3 mg/kg, s.c.) and NOR3 (0.1 mg/kg, s.c.), increased the NO contents in gastric mucosa. Cimetidine (100 mg/kg, p.o.), ranitidine (30 mg/kg, p.o.) and famotidine (10 mg/kg, p.o.) did not significantly affect the NO production. Pretreatment with NG-nitro-L-arginine methyl ester (3 mg/kg, i.v.), a NO synthase inhibitor, decreased the level of IT-066 (3 mg/kg, p.o.)-stimulated NO production. These results suggest that IT-066 stimulated gastric mucosal NO production, and that endogenous NO may be implicated in the pharmacological effect of IT-066. PMID- 9730009 TI - Investigation of the molecular changes during chemotherapy in non-Hodgkin's lymphoma. AB - In the present study the changes in the detection rate of bcl-2 and IgH gene rearrangements in relation to chemotherapy and therapeutic response in patients with diffuse large B-cell lymphoma have been investigated. Immunoglobulin gene rearrangements were detected in almost all patients during all stages of treatment. Persistence of bcl-2 rearrangements reflected the effect of chemotherapy better. Bcl-2 rearrangements were initially detected in 64% of the patients. Cells bearing the translocation disappeared during therapy in a significant group of cases. In 10 patients bcl-2-rearranged cells were detected for varying periods of time. However, no correlation was found between the molecular persistence or disappearance of cells as detected by PCR and the therapeutic response or recurrence rates. PMID- 9730008 TI - Soluble intercellular adhesion molecule-1 and natural killer cell activity in gastric cancer patients. AB - Intercellular adhesion molecule-1 (ICAM-1), a molecule bound to the cell surface, is a ligand for leukocyte function antigen-1 (LFA-1), and the ICAM-1/LFA-1 system mediates various cell-cell interactions involved in immunity. Soluble ICAM-1 (sICAM-1) is a circulating substance and binds with LFA-1 of leukocytes, thus, making leukocytes less available for binding with cell surface ICAM-1 on target cells. The serum level of soluble ICAM-1 (sICAM-1) was found to be significantly elevated (p<0.01) in patients with early and advanced gastric cancer compared with healthy controls. Natural killer activity (NK activity) was assessed by measuring the cytotoxicity of peripheral blood mononuclear cells (PBMCs) for K562 cells. There was no significant difference in NK activity between gastric cancer patients and healthy controls when heat-inactivated fetal calf serum was used in assays. However, addition of patient serum significantly decreased (p<0.05) NK activity when the serum was from patients with advanced gastric cancer compared with healthy volunteers. Addition of anti-ICAM-1 monoclonal antibody 0 to 5.0 microg/ml caused little change in NK activity in healthy controls, but its addition at 10 microg/ml remarkably decreased NK-activity in gastric cancer patients, probably through antibody binding with ICAM-1 on target cells. In other experiments, liver metastasis was induced in mice by inoculation of colon 26 murine colon cancer cells. In vitro pretreatment of colon 26 cells with the anti ICAM-1 monoclonal antibody significantly increased the number of metastatic nodules. These results suggest that both sICAM-1 and anti-ICAM-1 monoclonal antibody act as immunosuppressive factors by inhibiting the ICAM-1/LFA-1 system. PMID- 9730010 TI - In vitro cytotoxic activity of 1-decarboxy-3-oxo-ceanothic acid in a human ovarian adenocarcinoma cell line. AB - The effect of a novel pentacyclic triterpene, 1-decarboxy-3-oxo-ceanothic acid (DOCA) on DNA synthesis, DNA degradation and programmed cell death was examined in human ovarian adenocarcinoma (OVCAR-3) cells. OVCAR-3 cells exposed to various concentrations of DOCA for 30 h displayed a dose-dependent inhibition of DNA synthesis. Morphologically, treatment with 10 microg/ml of DOCA for 24 h and 72 h resulted respectively in reduction in cell volume and condensation of nuclear structures. By agarose gel analysis, DNA fragmentation with the characteristic pattern of inter-nucleosomal ladder was observed after cells were treated with 2.5 microg/ml of DOCA for 24 h. Both cell death and DNA fragmentation caused by this compound were partially inhibited by the protein synthesis inhibitor cycloheximide, suggesting that the apoptotic process caused by DOCA requires synthesis of new proteins. On the other hand, no apparent double-stranded DNA breaks were detected after cells were incubated with 2.5 microg/ml of DOCA for 24 h, indicating that DNA damage was not a preceding event for apoptosis induced by this compound. Taken together, our results demonstrate that the cytotoxic effect of DOCA is mediated, at least in part, by the induction of apoptosis. PMID- 9730011 TI - Impaired expression of atrial natriuretic peptide in diabetic rats with myocardial infarction. AB - We evaluated the effects of myocardial infarction (MI) on the hemodynamics and the expression of atrial natriuretic peptide (ANP) mRNA in rats with streptozotocin-induced diabetes. Eight weeks after streptozotocin injection, the diabetic rats and age-matched nondiabetic controls underwent coronary artery ligation. One week later, the left ventricular end-diastolic pressure, systolic blood pressure, infarct size, and serum ANP levels did not differ significantly between the diabetic and nondiabetic rats. Compared with control animals without MI, the atrial ANP/beta-actin mRNA ratio in rats with MI was increased to 195% in diabetic animals and 213% in nondiabetic animals. In the left ventricle, however, the ANP/beta-actin mRNA ratio in diabetic animals with MI was increased to only 131% compared with control animals, whereas the corresponding increase in nondiabetic animals was 240% (p<0.05). Thus, the modulation of ANP mRNA expression after MI was impaired in the left ventricle, but not in the atria, of diabetic rats. A reduced myocardial expression of ANP could increase the morbidity and mortality associated with cardiovascular disorders in patients with diabetes. PMID- 9730012 TI - Role of potassium channels in halothane-epinephrine arrhythmias. AB - It has been reported that antiarrhythmic drugs possessing the property of potassium channel blockade were most effective in preventing halothane epinephrine induced arrhythmias. Recent attention has focused on ATP-sensitive potassium (K(ATP)) channels because of their contribution to the cardiovascular actions of volatile anesthetics. The present study was designed to evaluate whether K(ATP) channels or transient outward potassium channels (Ito) were involved in the mechanism of halothane-epinephrine arrhythmias in rat. Rats were anesthetized with halothane (1.5%), and the lungs were mechanically ventilated. The arrhythmogenic thresholds of epinephrine during halothane anesthesia were determined in 74 rats receiving saline or one of tested agents. The arrhythmogenic dose of epinephrine (ADE) was significantly increased by a K(ATP) channel opener, JTV506 (P < 0.01), and had a tendency to be increased by other K(ATP) channel openers, cromakalim, nicorandil, KRN2391 and Y 26763, but were not affected by a K(ATP) channel blocker, glibenclamide. The Ito blocker, 4 aminopyridine, also significantly increased the ADE. Epinephrine produced second degree or complete atrioventricular block in 4 out of 7 rats receiving glibenclamide. These results suggest that Ito but not K(ATP) channels might be involved in the mechanism in producing halothane-epinephrine arrhythmias. PMID- 9730014 TI - Data mining in finance: introducing the special issue of IJNS. PMID- 9730013 TI - Docosahexaenoic acid inhibits blood viscosity in stroke-prone spontaneously hypertensive rats. AB - Increased blood viscosity facilitates the formation of thrombosis. This is an important risk factor in the occurrence of cerebral infarctions. The present study was undertaken to elucidate whether docosahexaenoic acid (DHA) inhibits blood viscosity, hematocrit and fibrinogen in the disease animal model, stroke prone spontaneously hypertensive rats (SHRSP). An attempt was also made to clarify the effect of DHA on blood pressure in SHRSP. Blood viscosity, hematocrit and fibrinogen in non-treated SHRSP increased significantly when compared with levels in age-matched non-treated Wistar Kyoto rats (WKY). SHRSP rats which were administered DHA for 5 weeks displayed significant decreases in blood viscosity, hematocrit and fibrinogen when compared with the values in non-treated SHRSP. The blood pressure of DHA-treated SHRSP was significantly lower than that of non treated SHRSP. A positive correlation existed between blood pressure and blood viscosity. These findings suggest that decreased blood viscosity induced by DHA appears to be associated with the reduction of thrombosis formation and hypotensive action in SHRSP. PMID- 9730015 TI - Understanding time series networks: a case study in rule extraction. AB - A significant limitation of neural networks is that the representations they learn are usually incomprehensible to humans. We have developed an algorithm, called TREPAN, for extracting comprehensible, symbolic representations from trained neural networks. Given a trained network, TREPAN produces a decision tree that approximates the concept represented by the network. In this article, we discuss the application of TREPAN to a neural network trained on a noisy time series task: predicting the Dollar-Mark exchange rate. We present experiments that show that TREPAN is able to extract a decision tree from this network that equals the network in terms of predictive accuracy, yet provides a comprehensible concept representation. Moreover, our experiments indicate that decision trees induced directly from the training data using conventional algorithms do not match the accuracy nor the comprehensibility of the tree extracted by TREPAN. PMID- 9730016 TI - Modeling volatility using state space models. AB - In time series problems, noise can be divided into two categories: dynamic noise which drives the process, and observational noise which is added in the measurement process, but does not influence future values of the system. In this framework, we show that empirical volatilities (the squared relative returns of prices) exhibit a significant amount of observational noise. To model and predict their time evolution adequately, we estimate state space models that explicitly include observational noise. We obtain relaxation times for shocks in the logarithm of volatility ranging from three weeks (for foreign exchange) to three to five months (for stock indices). In most cases, a two-dimensional hidden state is required to yield residuals that are consistent with white noise. We compare these results with ordinary autoregressive models (without a hidden state) and find that autoregressive models underestimate the relaxation times by about two orders of magnitude since they do not distinguish between observational and dynamic noise. This new interpretation of the dynamics of volatility in terms of relaxators in a state space model carries over to stochastic volatility models and to GARCH models, and is useful for several problems in finance, including risk management and the pricing of derivative securities. Data sets used: Olsen & Associates high frequency DEM/USD foreign exchange rates (8 years). Nikkei 225 index (40 years). Dow Jones Industrial Average (25 years). PMID- 9730017 TI - A constrained neural network Kalman filter for price estimation in high frequency financial data. AB - In this paper we present a neural network extended Kalman filter for modeling noisy financial time series. The neural network is employed to estimate the nonlinear dynamics of the extended Kalman filter. Conditions for the neural network weight matrix are provided to guarantee the stability of the filter. The extended Kalman filter presented is designed to filter three types of noise commonly observed in financial data: process noise, measurement noise, and arrival noise. The erratic arrival of data (arrival noise) results in the neural network predictions being iterated into the future. Constraining the neural network to have a fixed point at the origin produces better iterated predictions and more stable results. The performance of constrained and unconstrained neural networks within the extended Kalman filter is demonstrated on "Quote" tick data from the $/DM exchange rate (1993-1995). PMID- 9730018 TI - Nonlinear trading models through Sharpe Ratio maximization. AB - While many trading strategies are based on price prediction, traders in financial markets are typically interested in optimizing risk-adjusted performance such as the Sharpe Ratio, rather than the price predictions themselves. This paper introduces an approach which generates a nonlinear strategy that explicitly maximizes the Sharpe Ratio. It is expressed as a neural network model whose output is the position size between a risky and a risk-free asset. The iterative parameter update rules are derived and compared to alternative approaches. The resulting trading strategy is evaluated and analyzed on both computer-generated data and real world data (DAX, the daily German equity index). Trading based on Sharpe Ratio maximization compares favorably to both profit optimization and probability matching (through cross-entropy optimization). The results show that the goal of optimizing out-of-sample risk-adjusted profit can indeed be achieved with this nonlinear approach. PMID- 9730019 TI - Using a financial training criterion rather than a prediction criterion. AB - The application of this work is to decision making with financial time series, using learning algorithms. The traditional approach is to train a model using a prediction criterion, such as minimizing the squared error between predictions and actual values of a dependent variable, or maximizing the likelihood of a conditional model of the dependent variable. We find here with noisy time series that better results can be obtained when the model is directly trained in order to maximize the financial criterion of interest, here gains and losses (including those due to transactions) incurred during trading. Experiments were performed on portfolio selection with 35 Canadian stocks. PMID- 9730020 TI - Switching portfolios. AB - A constant rebalanced portfolio is an asset allocation algorithm which keeps the same distribution of wealth among a set of assets along a period of time. Recently, there has been work on on-line portfolio selection algorithms which are competitive with the best constant rebalanced portfolio determined in hindsight (Cover, 1991; Helmbold et al., 1996; Cover and Ordentlich, 1996). By their nature, these algorithms employ the assumption that high returns can be achieved using a fixed asset allocation strategy. However, stock markets are far from being stationary and in many cases the wealth achieved by a constant rebalanced portfolio is much smaller than the wealth achieved by an ad hoc investment strategy that adapts to changes in the market. In this paper we present an efficient portfolio selection algorithm that is able to track a changing market. We also describe a simple extension of the algorithm for the case of a general transaction cost, including the transactions cost models recently investigated in (Blum and Kalai, 1997). We provide a simple analysis of the competitiveness of the algorithm and check its performance on real stock data from the New York Stock Exchange accumulated during a 22-year period. On this data, our algorithm outperforms all the algorithms referenced above, with and without transaction costs. PMID- 9730021 TI - Improved option pricing using artificial neural networks and bootstrap methods. AB - A hybrid neural network is used to predict the difference between the conventional option-pricing model and observed intraday option prices for stock index option futures. Confidence intervals derived with bootstrap methods are used in a trading strategy that only allows trades outside the estimated range of spurious model fits to be executed. Whilst hybrid neural network option pricing models can improve predictions they have bias. The hybrid option-pricing bias can be reduced with bootstrap methods. A modified bootstrap predictor is indexed by a parameter that allows the predictor to range from a pure bootstrap predictor, to a hybrid predictor, and finally the bagging predictor. The modified bootstrap predictor outperforms the hybrid and bagging predictors. Greatly improved performance was observed on the boundary of the training set and where only sparse training data exists. Finally, bootstrap bias estimates were studied. PMID- 9730022 TI - A first application of independent component analysis to extracting structure from stock returns. AB - This paper explores the application of a signal processing technique known as independent component analysis (ICA) or blind source separation to multivariate financial time series such as a portfolio of stocks. The key idea of ICA is to linearly map the observed multivariate time series into a new space of statistically independent components (ICs). We apply ICA to three years of daily returns of the 28 largest Japanese stocks and compare the results with those obtained using principal component analysis. The results indicate that the estimated ICs fall into two categories, (i) infrequent large shocks (responsible for the major changes in the stock prices), and (ii) frequent smaller fluctuations (contributing little to the overall level of the stocks). We show that the overall stock price can be reconstructed surprisingly well by using a small number of thresholded weighted ICs. In contrast, when using shocks derived from principal components instead of independent components, the reconstructed price is less similar to the original one. ICA is shown to be a potentially powerful method of analyzing and understanding driving mechanisms in financial time series. The application to portfolio optimization is described in Chin and Weigend (1998). PMID- 9730023 TI - Disinfection by-products and adverse pregnancy outcomes: what is the agent and how should it be measured? PMID- 9730024 TI - Does blindness protect against cancers? PMID- 9730025 TI - Exposure to trihalomethanes and adverse pregnancy outcomes. AB - Exposure during pregnancy to disinfection by-products in drinking water has been hypothesized to lead to several adverse reproductive outcomes. We performed a retrospective cohort study to examine the relation of trihalomethane exposure during the third trimester of pregnancy to low birthweight, term low birthweight, and preterm delivery. We matched Colorado birth certificates from January 1, 1990, through December 31, 1993, to historical water sample data with respect to time and location of maternal residence based on census block groups. After excluding births from all census block groups with no trihalomethane sample data and restricting to singleton white births with 28-42 weeks of completed gestation (>400 gm), we studied 1,893 livebirths within 28 census block groups. We found a weak association of trihalomethane exposure during the third trimester with low birthweight (odds ratio = 2.1 for the highest exposure level; 95% confidence interval = 1.0-4.8); a large increase in risk for term low birthweight at the highest level of exposure (odds ratio = 5.9; 95% confidence interval = 2.0-17.0); and no association between exposure and preterm delivery (odds ratio = 1.0 for the highest exposure level; 95% confidence interval = 0.3-2.8). The small number of adverse outcomes reduced the precision of risk estimates, but these data indicate a potentially important relation between third trimester exposure to trihalomethanes and retarded fetal growth. PMID- 9730026 TI - Reduced cancer incidence among the blind. AB - Melatonin is a hormone primarily produced by the pineal gland at night and is suppressed by exposure to light. Experimental studies have indicated that melatonin may protect against cancer development. In the majority of totally blind people, melatonin is never suppressed by light exposure. The aim of this study was to test the hypothesis that blind people have a decreased cancer incidence, and that this effect is more pronounced in the totally blind than in the severely visually impaired. We identified a cohort of 1,567 totally blind and 13,292 severely visually impaired subjects and obtained information about cancer incidence from the Swedish Cancer Registry. We calculated standardized incidence ratios (SIRs) based on the number of person-years and incidence rates specific for national age, sex, and calendar year. Totally blind people had a lower incidence of all cancers combined [SIR = 0.69; 95% confidence interval (CI) = 0.59-0.82]. The risk reduction was observed in both men and women and was equally pronounced in hormone-dependent tumors as in other types of cancer. In the severely visually impaired, SIR was 0.95 (95% CI = 0.91-1.00). The findings support the hypothesis that blind people have a lower cancer incidence, although other explanations than the higher melatonin exposure must also be considered. PMID- 9730027 TI - Time-series analysis of air pollution and cause-specific mortality. AB - Ten large European cities provided data on daily air pollution as well as mortality from respiratory and cardiovascular mortality. We used Poisson autoregressive models that controlled for trend, season, influenza epidemics, and meteorologic influences to assess the short-term effects of air pollution at each city. We then compared and pooled the city-specific results in a meta-analysis. The pooled relative risks of daily deaths from cardiovascular conditions were 1.02 [95% confidence interval (CI) = 1.01-1.04] for a 50 microg/m3 increment in the concentration of black smoke and 1.04 (95% CI = 1.01-1.06) for an increase in sulfur dioxide levels in western European cities. For respiratory diseases, these figures were 1.04 (95% CI = 1.02-1.07) and 1.05 (95% CI = 1.03-1.07), respectively. These associations were not found in the five central European cities. Eight-hour averages of ozone were also moderately associated with daily mortality in western European cities (relative risk = 1.02; 95% CI = 1.00-1.03 for cardiovascular conditions and relative risk = 1.06; 95% CI = 1.02-1.10 for respiratory conditions). Nitrogen dioxide did not show consistent relations with daily mortality. These results are similar to previously published data and add credence to the causal interpretation of these associations at levels of air pollution close to or lower than current European standards. PMID- 9730028 TI - Characteristics related to the prevalence of minimal or mild endometriosis in infertile women. Canadian Collaborative Group on Endometriosis. AB - The objective of this case-control study is to identify factors associated with the prevalence of minimal or mild endometriosis among infertile women. Cases (N = 329) were women diagnosed by laparoscopy with minimal or mild endometriosis and without any other factors explaining their infertility. Controls (N = 262) were women in whom the infertility remained unexplained after a diagnostic laparoscopy. Selected characteristics were documented by means of a face-to-face interview before the laparoscopy. The prevalence of minimal or mild endometriosis was higher in women age 25 years or older, in those who reported menarche at the age of 13 years [prevalence odds ratio (POR) = 1.63; 95% confidence interval (CI) = 1.02-2.60] or older (POR = 1.73; 95% CI = 1.07-2.78), menstrual cycles of 27 days or less (POR = 1.63; 95% CI = 1.02-2.60), or caffeine intake of 300 mg per day or more (POR = 1.33; 95% CI = 0.91-1.94). The prevalence of minimal or mild endometriosis was inversely related to body mass index. Parous women were less likely to have endometriosis (POR = 0.61; 95% CI = 0.39-0.96) than were nulliparous women. Education, duration of infertility, and smoking status were not related to the presence of endometriosis. PMID- 9730029 TI - Risk of uterine leiomyomata among premenopausal women in relation to body size and cigarette smoking. AB - To investigate whether factors influencing ovarian function affect risk of uterine leiomyomata, we examined prospectively the association of new diagnoses confirmed by ultrasound or hysterectomy with body mass index, weight change, height, and cigarette smoking among 94,095 premenopausal women with intact uteri, who were ages 25-42 years at the start of follow-up in 1989. We assessed body mass index and cigarette smoking from responses on the study questionnaire completed just before diagnosis. During 322,775 person-years, 2,967 new cases of uterine leiomyomata confirmed by ultrasound or hysterectomy were reported. Risk among all cases confirmed by ultrasound or hysterectomy increased with increasing adult body mass index. The multivariate relative risks (RR) and 95% confidence intervals (CI) according to body mass index categories of <20.0, 20.0-21.9, 22.0 23.9, 24.0-25.9, 26.0-27.9, 28.0-29.9, and > or =30.0 were 0.90 (95% CI = 0.79 1.03), 1.00 (referent), 1.08 (95% CI = 0.97-1.21), 1.16 (95% CI = 1.03-1.31), 1.21 (95% CI = 1.05-1.40), 1.36 (95% CI = 1.16-1.59), and 1.23 (95% CI = 1.09 1.39), respectively. The RRs for hysterectomy-confirmed cases generally were higher. Similarly, risk was positively associated with weight gain since age 18 years. Body mass index at age 18 years, height, and cigarette smoking were unrelated to risk of uterine leiomyomata. Elevated adult body mass index is associated with a modest increased risk of uterine leiomyomata among premenopausal women. PMID- 9730030 TI - A case study comparing a meta-analysis and a pooled analysis of studies of sinonasal cancer among wood workers. AB - A pooled analysis of raw data from 12 case-control studies of sinonasal cancer has recently been conducted by the International Agency for Research on Cancer; the summary odds ratio for all wood-related occupations was 2.0 (95% confidence interval = 1.6-2.5). We have conducted a meta-analysis of the published results of 14 studies on the same topic, including 11 studies from the pooled analysis, and compared the results for several categories of wood workers. Usable results were available for 12 studies: male wood workers had a summary odds ratio of sinonasal cancer of 2.6 (95% confidence interval = 2.1-3.3). The corresponding value based only on the studies that were also included in the pooled analysis was 2.4 (95% confidence interval = 1.9-3.0). When our meta-analysis was based on a relatively large number of studies, results tended to be higher than those of the pooled analysis. As compared with the meta-analysis, the pooled analysis controlled the effect of publication bias by using data from studies for which no specific result was reported, and it reduced misclassification of exposure; the resources used in the pooled analysis, however, were one order of magnitude greater than those needed for the meta-analysis. Whether a pooled analysis of raw data or a meta-analysis should be carried out depends on the research question; we propose some criteria for this decision. PMID- 9730031 TI - Family history of breast cancer as a predictor for fatal prostate cancer. AB - To examine the relation between family history of breast cancer in a mother or sister and a man's risk of fatal prostate cancer, we analyzed data from a prospective mortality study of adult men in the United States. During 12 years of follow-up, there were 3,141 deaths from prostate cancer in a cohort of 480,802 men who were cancer-free at study entry in 1982. Results from Cox proportional hazards models, adjusted for other risk factors, showed a modest increased risk of fatal prostate cancer associated with a family history of breast cancer (in the absence of a family history of prostate cancer) [rate ratio (RR) = 1.16; 95% confidence interval (CI) = 1.01-1.33]. The association was stronger among men younger than 65 years of age whose relatives were diagnosed with breast cancer before age 50 years (RR = 1.65; 95% CI = 0.88-3.10) and among Jewish men (RR = 1.73; 95% CI = 1.00-2.97). The increased risks observed in these subgroups may reflect genetic alterations underlying familial clustering of prostate and breast cancer. PMID- 9730032 TI - Do education and income buffer the effects of death of spouse on mortality? AB - In this paper, we estimate the effects of the death of a spouse on the mortality of the survivor in different education and income groups. These socioeconomic resources may buffer the harmful effects of the stressful life event of the loss of one's spouse. The data come from a prospective study of mortality among all 35 to 74-year-old married Finnish persons. Follow-up was established by record linkage to death certificate registers for 1986-1991 (about 86,000 deaths, of which almost 5,500 occurred among the bereaved). The relative mortality after the death of one's spouse was broadly similar in different education and income groups. Absolute differences in mortality rates between bereaved and nonbereaved persons were larger in the lower end of the social spectrum, however. This pattern held for four broad categories of death: both sexes and two age groups (35-64 years and 65-74 years). The degree to which socioeconomic resources buffer the effects of death of spouse depends on whether it is assessed in terms of rate differences or rate ratios. Nevertheless, regardless of measurement choice, the effects of bereavement exist in all socioeconomic groups analyzed in this study. Furthermore, because of the high absolute level of mortality, the burden of excess mortality experienced after the death of one's spouse is heavier in the lower social strata. PMID- 9730033 TI - Vitamin D receptor genotype and the risk of bone fractures in women. AB - Several studies have confirmed an initial report of a relation between bone density and polymorphic forms of the calcitriol (vitamin D) receptor gene, whereas others have failed to find an association. We examined whether variants of the vitamin D receptor gene are associated with the risk of bone fracture, using a nested case-control analysis within the Nurses' Health Study cohort. The study women all were Caucasian and were 43-69 years of age when they provided a blood sample. Cases included the 54 proximal femur (hip) fractures and 163 distal radius (forearm) fractures that occurred subsequent to the blood draw. Cases and controls were genotyped by polymerase chain reaction for the BsmI polymorphism. The BB genotype, previously associated with lower bone density, was associated with a more than twofold increased risk of hip fracture compared with the bb genotype. Risk was greater for women who were older, leaner, or less physically active or who had a lower calcium intake. The heterozygous genotype was not associated with any increased risk of hip fracture, and we observed little association between vitamin D receptor genotype and forearm fracture. This study supports an association between vitamin D receptor genotype and hip fracture. It also implies that modification by other risk factors may have contributed to the conflicting results from previous studies of vitamin D receptor genotype and femoral bone density. PMID- 9730035 TI - Parental smoking and infection with Helicobacter pylori among preschool children in southern Germany. AB - Exposure to parental smoking is known to increase children's susceptibility to a variety of infections. We investigated the relation of parental smoking to infection with Helicobacter pylori in a population-based study among preschool children who were screened for school fitness in the city of Ulm, Germany, in 1996. Current infection with H. pylori was measured by a 13C-labeled urea breath test. Information on factors potentially related to H. pylori infection, including parental smoking in the household, was obtained from children's parents through a self-administered standardized questionnaire. Among 1,201 eligible children, 945 participated in the study (response rate = 79%). Overall prevalence of active infection was 13.7%. After adjustment for confounding factors, we found a strong positive relation between smoking by the father in the household and H. pylori infection (odds ratio = 3.7; 95% confidence interval = 2.3-6.1). By contrast, there was a strong negative relation between smoking by the mother and H. pylori infection (odds ratio = 0.4; 95% confidence interval = 0.2-0.8) that was most pronounced among children who had been breastfed. These striking patterns cannot be explained by current knowledge. PMID- 9730034 TI - Microsomal epoxide hydrolase polymorphism and risk of spontaneous abortion. AB - We conducted a nested case-control study to examine the association of microsomal epoxide hydrolase polymorphism with spontaneous abortion. The analysis included 127 cases and 107 controls from a rural community in China. The prevalence of the homozygous wild-type (AA), the heterozygous variant (Aa), and the homozygous variant (aa) in exon 3 of epoxide hydrolase was 13.4%, 34.6%, and 52.0% in cases and 27.1%, 30.8%, and 42.1% in controls, respectively. In contrast, the variant genotypes in exon 4 of epoxide hydrolase were less frequent in cases than controls. Using women with genotype AA as the referent, the adjusted odds ratio of spontaneous abortion was 2.69 [95% confidence interval (CI) = 1.33-5.42] for those with genotype Aa or aa in exon 3; but it was 0.45 (95% CI = 0.22-0.94) for those with genotype Aa or aa in exon 4, indicating that the two variants have opposite associations with spontaneous abortion. The findings persisted after adjustment for age, education, parity, smoking, alcohol use, occupation, and pesticide exposure, as well as in subgroup analysis. Moreover, for the variant genotypes Aa or aa in exon 3, the odds ratio was twice as great in those cases with three or more spontaneous abortions than in those with fewer spontaneous abortions. PMID- 9730036 TI - Cardiovascular diseases and diabetes mellitus in different ethnic groups: the Finnmark study. AB - The mortality from coronary and cerebrovascular diseases is higher in Finnmark County than in other Norwegian counties. In a population-based cohort study, we compared the incidence of myocardial infarction, stroke, and diabetes mellitus in different ethnic groups in Finnmark. A total of 10,622 subjects of Norse, Sami, and Finnish origin were followed for 14 years. During approximately 150,000 person-years, we identified 509 and 84 cases of myocardial infarction, 107 and 75 cases of stroke, and 96 and 73 cases of clinical diabetes mellitus among men and women, respectively. A total of 533 men and 199 women died. Norse subjects born outside of Finnmark had the most favorable risk factor levels and, in general, the lowest incidence of disease. Men of Finnish origin had a higher incidence rate of all endpoints than other men, and Finnish women had a higher incidence rate of myocardial infarction than other women. Sami women were more obese but did not have a higher diabetes mellitus incidence than other women. After adjustment for major cardiovascular risk factors and height, most ethnic differences were attenuated. PMID- 9730037 TI - Modeling disease incidence rates in families. AB - We apply an extended Cox model to study latent genes and measured environmental exposures simultaneously as risk factors for disease. Using this method, we assume Mendelian transmission of the genes and either dominant or recessive gene action. We compared the results from this model with those obtained under a model that includes the environmental variables and a family risk score. We demonstrate the method in samples of 1,433 Caucasian families (N = 6,791) and 206 African American families (N = 771) from the National Heart, Lung, and Blood Institute Family Heart Study. In Caucasians, we found evidence suggesting that having ever smoked increased the risk of coronary heart disease only in individuals who carry a genetic susceptibility. We also noted that in both Caucasian and African American families, the relative risk of coronary heart disease for ever-treated vs never-treated for high serum total cholesterol increased after including an unobserved susceptibility genotype in the model. This finding implied that there may be genes influencing coronary heart disease independent of those that influence total cholesterol. Such findings were not evident when genetic risk was summarized by the family history score. We also discuss the extension of the model to address the etiology of complex diseases. PMID- 9730038 TI - Software for hierarchical modeling of epidemiologic data. AB - Hierarchical models can provide more reasonable and stable parameter estimates than conventional analytical approaches. This technique also deals with problems of multiple comparisons and allows one to model multilevel data within a hierarchical framework. Hence, one would anticipate a surge in applying hierarchical models to epidemiologic data. Difficulties in fitting hierarchical models, however, seem to have limited their use. To help address this problem, we describe the existing software packages that one can use to fit hierarchical models. Since these packages have limited familiarity and applicability in epidemiology, we also present SAS code for analyzing epidemiologic data with hierarchical models. These results allow epidemiologists to fit hierarchical models with readily available software. PMID- 9730039 TI - Measurement error from assessing use of vitamin supplements at one point in time. AB - Although many epidemiologic studies ask about current use of vitamin supplements, long-term use is usually the exposure of etiologic interest. We conducted a mailed survey to investigate the relation between current and long-term (10-year) supplement use (N = 325 adults). Estimates of current daily intake for supplemental micronutrients were roughly twice that of average daily intake over the past 10 years. Correlations between current intake and long-term intake from supplements alone were 0.77, 0.75, and 0.65 for vitamin C, vitamin E, and calcium, respectively. A measure of supplement use at one point in time incorporates measurement error that will attenuate measures of association. PMID- 9730040 TI - Insulin-like growth factor-I in relation to premenopausal ductal carcinoma in situ of the breast. AB - We evaluated the association of plasma insulin-like growth factor-I (IGF-I) and IGF binding protein-3 (IGFBP-3) with risk of breast cancer in a study of 94 cases of premenopausal ductal carcinoma in situ and 76 controls. Compared with women in the lowest tertile of IGF-I, women in the upper two tertiles of IGF-I had an elevated risk for ductal carcinoma in situ. Conversely, compared with women in the lowest tertile of IGFBP-3, women in the upper two tertiles of IGFBP-3 had a decreased risk for ductal carcinoma in situ. After grouping women on the basis of both IGF-I and IGFBP-3, women in the highest two tertiles of IGF-I and the lowest tertile of IGFBP-3 were at notably higher risk than women in the lowest tertile of IGF-I and the highest two tertiles of IGFBP-3 (odds ratio = 3.7; 95% confidence interval = 1.1-12.2). We conclude that the combination of high IGF-I and low IGFBP-3 may increase the risk of premenopausal ductal carcinoma in situ. PMID- 9730041 TI - Is geographic variation in hip fracture rates related to current or former region of residence? AB - This study examines geographic variation in male and female age-adjusted hip fracture rates in white elderly Medicare enrollees. We assembled a cohort of more than 2 million 1992 enrollees and followed them passively through record linkage for 2 years for a hospitalization containing a diagnostic code indicating hip fracture. We simultaneously estimated rate ratios by region of residence early (inferred from the Social Security number) and late in life. Residence region early in life is associated with substantial variation in hip fracture rates. Conversely, current region has little relation with hip fracture risk. PMID- 9730042 TI - Scientific paradigms in epidemiology and professional values. PMID- 9730043 TI - Urinary tract infections and condoms. PMID- 9730044 TI - Maternal alcohol and caffeine consumption and spontaneous abortion. PMID- 9730045 TI - Validation of Norwegian death certificates on dementia in residents of nursing homes. PMID- 9730046 TI - Oligosaccharide organization of the beta-subunits of pig kidney Na+/K+-ATPase. AB - The distance between the beta-subunits of Na+/K+-ATPase isolated from pig dark red kidney medulla was determined by Forster energy transfer. First, oligosaccharides of the beta-subunit were shown to be labelled with three fluorophores: Lucifer yellow (LY), Lissamine rhodamine B sulfonyl hydrazine (LRSH) and Cascade blue (CB). Further, LY and LRSH were used as the donor and the acceptor, respectively, for Forster energy transfer studies to determine the localization of the beta-subunit in the native enzyme which is known to be formed as a tetramer (alphabeta)2. It was found that the beta-subunits in the functional enzyme complex in the membrane are not localized next to each other but are spatially separated. The distance between fluorophores covalently attached to the beta-subunits was found to be 5.1 nm. This conclusion was confirmed by measurements with another donor-acceptor pair CB-LY. The results also support the idea of a direct interaction of the beta-subunit with the extracellular part of the alpha-subunit. These interactions were modified in the presence of millimolar concentrations of magnesium ions. This indicates a crucial role of magnesium in extracellular interactions between the alpha and beta subunits. PMID- 9730047 TI - Effect of captopril on the development of left ventricular hypertrophy in rabbits with aortic insufficiency. AB - We investigated the effect of captopril on the growth of the left ventricle in an experimental model of aortic insufficiency. Four groups of rabbits were studied 28 days after experimental intervention: 1. control, 2. control with captopril (10 mg/kg/day), 3. aortic insufficiency, 4. aortic insufficiency with captopril (10 mg/kg/day). Aortic insufficiency induced hypertrophic growth of the left ventricle demonstrated by increased weight and ribonucleic acid (RNA) concentration. Administration of captopril only slightly attenuated the weight increase of the left ventricle and the increase in concentration of left ventricular RNA. However, captopril reduced the concentration of left ventricular deoxyribonucleic acid (DNA) both in the control and even more in the group with aortic insufficiency. The chronic haemodynamic overload enhanced mitochondrial respiration in the left ventricle which was not influenced by captopril. We conclude that captopril in the dose 10 mg/kg/day did not prevent hypertrophy of the left ventricle but reduced left ventricular DNA concentration. PMID- 9730048 TI - Effect of chronic renal insufficiency on the function and metabolic parameters of the isolated rat heart. AB - Chronic renal insufficiency (CRI) is often associated with cardiovascular disease; however, its underlying mechanisms are not completely understood. Therefore, in the present study, myocardial functions and metabolic changes were investigated using an animal model of CRI in subtotally nephrectomized rats. In addition, some other parameters, considered risk factors of cardiovascular diseases, were determined. Subtotal nephrectomy led to an elevation in blood pressure (144 +/- 2.8 vs 114 +/- 2.5 mm Hg), left ventricular hypertrophy (290 +/ 12 vs 200 +/- 40 mg/100 g b.w.), hypertriglyceridaemia (2.96 +/- 0.31 vs 0.77 +/ 0.07 mmol/l), and impaired glucose tolerance (AUC 836 +/- 12.4 vs 804 +/- 10.4 mmol x l(-1) x 120 min). Isolated perfused hearts of uraemic rats exhibited diminished basal functions (coronary and aortic flow, stroke volume) by 20-30% compared with the controls. Interestingly, the tolerance of isolated heart to global 20-min no-flow ischaemia was improved in uraemic rats. The most marked differences in heart function recovery during reperfusion concerned aortic flow (90 +/- 2.3 vs 66 +/- 10%) and stroke volume (97 +/- 2.7 vs 68 +/- 5.6% of pre ischaemic values). Pre-ischaemic myocardial glycogen content was distinctly increased (by 50%) in uraemic rats compared with the controls. PMID- 9730049 TI - Lactate dehydrogenase and its isoenzymes in the marrow and peripheral blood from haematologically normal subjects. AB - Lactate dehydrogenase (LDH) activity and its isoenzymatic fractions were measured in bone marrow blood and in peripheral venous blood from 16 haematologically normal subjects. Total LDH activity was significantly higher in marrow than in venous blood (428.8 +/- 98.4 vs 260.1 +/- 40.2 mU/l, p < 0.0001). The same was true for the absolute values of its isoenzymatic fractions. The percentage fractions LDH 1 and LDH 5 were similar in the two regions, while LDH 3 and LDH 4 were higher in medullary blood (p < 0.05) and LDH 2 was higher in peripheral blood (p < 0.05). The Spearman test showed a limited correlation between marrow and peripheral total LDH activity values (p < 0.05). This seems to be at least in part sustained by the highly significant correlations existing in LDH 3 and LDH 4 values, reported to be pre-eminent isoforms in maturing haematopoietic cells (p < 0.005 and p < 0.001, respectively). These findings could be attributed to an apoptotic regulation of marrow cell production. PMID- 9730050 TI - Effect of terguride on insulin binding, insulinaemia, glucose tolerance and hyperlipaemia in lean SHR Koletsky rats. AB - Glucose tolerance, insulin binding to erythrocytes, insulinaemia, plasma total cholesterol, plasma triglycerides, weight of fat pads, food consumption and body weight changes were studied in genetically hypertensive lean Koletsky rats. Long term treatment with dopaminergic agonist terguride (0.2 mg/kg/day) normalized glucose tolerance and increased the percentage of bound insulin to erythrocytes in both sexes. Terguride decreased insulinaemia, cholesterolaemia, fat pads and body weight only in female rats. Food consumption was not influenced by terguride over the injection period. PMID- 9730051 TI - Circadian oscillations of serum thyroid hormones in the laboratory rat: the effect of photoperiods. AB - The seasonal influence on circadian oscillations of serum thyroid hormones has been confirmed in the laboratory rat, an animal exhibiting low photoperiodic activity. The aim of this paper was to study the influence of various photoperiods, applied in a single season, on circadian variations in the levels of thyroid hormones in male Wistar rats. After 6-weeks of adaptation to artificial light-dark regimens (LD) 08:16 h, 12:12 h, 16:08 h, and to the standard housing conditions, the rats were examined in 3 h intervals in the course of 24 h in December. The concentrations of thyroxine (T4), triiodothyronine (T3) and reverse T3 (rT3) were examined in the serum. The curves of T4 circadian oscillations showed two peaks in all the photoperiods followed. Computative acrophases were localized between 07.00 and 08.00 h, the amplitude in the LD 12:12 regimen was twice that observed in LD 08:16 and 16:08, the rhythm was present and the mesors were approximately the same. Circadian oscillations of T3 exhibited rhythmicity in all the photoperiods with computative acrophases localized between 07.30 and 09.00 h, and the values of mesors in LD 08:16 and 16:08 regimens were significantly lower in comparison with those in the LD 12:12 regimen. The rT3 circadian variations in the LD 12:12 regimen showed rhythmicity with acrophase at 06.00 h. The rhythm in the LD 16:08 regimen was of borderline significance, the computative acrophase occurred at 8.16 h, and the mesor value was significantly higher than those in the LD 12:12 regimen. The decrease in the amplitude of T4 oscillations and the lower T3 mesors in LD 08:16 and 16:08 regimens in comparison with the LD 12:12 values indicated only minor modification in circadian oscillations of T4 and T3 resulting from artificial photoperiods. In comparison with our previous studies these data suggest that changes in circadian oscillations of serum thyroid hormones might reflect the effect of the season of the year rather than the effect of day duration, i.e. the photoperiod. PMID- 9730052 TI - Exposure to stress alters the effects of dynorphins in the hot plate test. AB - The analgesic effects of intracerebroventricularly (i.c.v.) administered dynorphin A(1-13) and its analog dynorphin A(1-10)amide using the hot plate test were studied in mice. Both dynorphins applied i.c.v. by the freehand method had an analgesic effect but no effect was seen when applied i.c.v. through an implanted cannula. Moreover, freehand i.c.v. injection of saline increased the time of immobility in the forced swimming test and glycaemia levels compared with intact mice. In contrast to the freehand injection, saline administration through an implanted cannula did not influence the immobility of animals in forced swimming test when compared with the intact controls. These results suggest that 1) the freehand method is very stressful procedure of administration which could influence the effects of dynorphins in the hot plate test and 2) dynorphins exert an analgesic effect in the hot plate test only when combined with a stressor (freehand i.c.v. injection). PMID- 9730054 TI - Reciprocal interactions between intralaminar and lateral thalamic nuclei in rats. AB - Reciprocal interactions between intralaminar thalamic nuclei (ncl. centralis lateralis, CL, and ncl. parafascicularis, Pf), the pretectal area (Pt) and lateral thalamic nuclei (ventrobasal complex, VB, ncl. anterior ventralis, AV, and ncl. ventralis anterior, VA) have been observed in ketamine-anaesthetized rats. Extracellular single unit activity has been recorded after single electrical stimuli. Electrical stimulation of the VB evoked a short latency orthodromic response followed by a pause in spontaneous activity in neurones of medial thalamic nuclei. Lateral thalamic neurones responded to electrical stimulation of the intralaminar nuclei or the pretectal area with the same pattern of response. Striatal, sensorimotor cortical or peripheral electrical stimulation also evoked similar responses. The pauses in spontaneous activity were shown to be the result of inhibition since the responsiveness of the intralaminar nuclei or the lateral thalamic neurones to all inputs was abolished or reduced after a conditioning electrical single-shock stimulation in the VB or in the intralaminar nuclei, respectively. The two components of the response were of a different origin, since most of the short latency responses disappeared after medullary, upper cervical sections or large decortications, while the inhibitions persisted. These inhibitions were shown to be of thalamic origin since their duration was decreased after extensive decortications increased after medullary section. It is concluded that the neuroneal properties studied in this report are probably broadly represented throughout the thalamus and that thalamic neurones are under inhibitory control elicited by afferent volleys. This inhibitory control includes a relay in the nucleus reticularis thalami (nRT). The mechanisms of sensory interaction can be purely thalamic, but they can be modulated by suprathalamic and/or mesencephalic loops. PMID- 9730053 TI - Some bronchoalveolar lavage parameters and leukocyte cytokine release in response to intratracheal instillation of short and long asbestos and wollastonite fibres in rats. AB - We investigated the differences between the lavage parameters -- including tumour necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) release by lavage leukocytes -- in control rats and in animals intratracheally instilled with short and long amosite and wollastonite fibres. These cytokines can play an important role in lung disease development after long-term exposure to some fibrous dusts. Short and long amosite and wollastonite fibres were intratracheally instilled in rats (1 mg/week) for ten weeks while saline was given to controls. To compare the harmful effects of these fibres, the number of leukocytes/ml of bronchoalveolar lavage (BAL), the number of alveolar macrophages (AM) per ml of BAL, AM:granulocyte (GR) ratios in lavage fluid, phagocytic activity and viability of AM, lactate dehydrogenase (LDH), acid phosphatase (AcP), and TNF-alpha and IFN-gamma release by lavage leukocytes were investigated 3 months after the first intratracheal instillation. Compared with the controls, amosite short fibres significantly decreased the numbers of AM/ml BAL, and increased their phagocytic activity and AcP release. Long amosite fibres significantly decreased the numbers of AM/ml BAL, increased the number of granulocytes depressed the phagocytic activity and viability of AM, and significantly decreased the levels of TNF-alpha and IFN-gamma in supernatants of cultured leukocytes. While wollastonite short and long fibre instillation did not significantly influence the parameters studied (except for a significantly increased number of leukocytes/ml BAL in wollastonite long fibres), amosite short and long fibres caused marked differences in these parameters, the long fibres being more effective. PMID- 9730055 TI - Abnormal neuroneal activities in intralaminar thalamic nuclei following chronic lesions of nucleus reticularis thalami in rats. AB - Extracellular single unit activity in the intralaminar thalamic nuclei (ncl. centralis lateralis, CL, n = 77 and ncl. parafascicularis, Pf, n = 163) and in the pretectal area (Pt, n = 75) was examined following chronic electrolytic lesions of the nucleus reticularis thalami (nRT) in ketamine-anaesthetized rats after single electrical stimuli to the ventrobasal complex (VB). Extensive alterations of either the ongoing ("spontaneous") activity or the pattern of VB evoked responses were observed. Four major changes were observed in the activity of these intralaminar or pretectal neurones: 1) many neurones were silent, two times more frequently than in a parallel study with control intact rats; 2) the firing pattern of all the other neurones was in the form of tonic (stationary like) discharge, without burst discharges as previously described in intact animals. They were ranked into classes according to their spontaneous discharge: class I, silent (no resting discharge) 12%, class II (1-15 Hz), 54 % and class III (> 16 Hz), 34%. Class III neurones were never found in intact rats; 3) electrical stimulation of the VB evoked a short latency orthodromic excitatory response in these neurones but this response was not followed by any slowing or depression of the spontaneous activity in more than 40% of recorded cells. When it occurred, this pause was shorter than that always observed in intact rats by more than 35% and longer in 7% of the responsive cells. All these changes were correlated with the extent of damage to the ipsilateral nRT; 4) VB stimulation evoked prolonged excitatory responses lasting more than 150 ms in 13% of the responsive cells, and nRT stimulation led to a short latency response followed by a pause of activity. These findings suggest that the nRT is involved in sensory integration and modulation. PMID- 9730056 TI - The expression of B-50/GAP-43 and GFAP after bilateral olfactory bulbectomy in rats. AB - In the present study we investigated the effect of a two-stage bilateral lesion of the olfactory bulb (OB) in rats on the regeneration ability of peripheral olfactory neurons and their reinnervation capacity in the spared OB. The outgrowth of newly-generated olfactory axons as well as the maturation of their terminal synaptic field was detected by immunohistochemistry of the growth associated phosphoprotein B-50/GAP-43. In addition, the glial response to the surgery was monitored by an immunohistochemical marker for astrocytes, glial fibrillary acidic protein (GFAP). In neonatal rats (P3-P5), the right OB was removed, then three months later the contralateral side was ablated. Six days after the second operation the animals were transcardially perfused. Their brains were embedded in paraplast, serially sectioned and processed for histological and immunohistochemical observations. After neonatal OB ablation, homogeneous B-50 immunoreactivity (BIR) was found in the forebrain, olfactory axons and ectopic glomeruli localized in the small OB remnant-like structures and in the regenerated neuroepithelium. A strong GFAP response was revealed in the brain cortex as well as in the newly-formed olfactory axons and glomeruli-like structures of the OB remnants. After adult OB ablation strong BIR was observed in olfactory axons, while remaining glomerular structures were only faintly stained. The neuroepithelium revealed signs of massive degenerative processes with a substantial decrease in BIR. The GFAP-positive astrocytes were scattered throughout the entire OB remnant and were prominent in the glomeruli-like structures and adjacent frontal cortex. In the present study, we applied GAP-43 and GFAP immunohistochemistry to characterize the responses of individual olfactory components after two-stage olfactory bulbectomy. Furthermore, this model of OB ablation characterized by two immunohistochemical markers could elucidate certain molecular mechanisms involved in the regeneration and/or plasticity of the olfactory system. PMID- 9730057 TI - Changes of the inner time structures in the sequences of interspike intervals produced by the activity of excitatory and inhibitory synapses: simulation with Gaussian input processes. AB - The computer simulation of space-temporal summation of post-synaptic potentials of neurone's membrane demonstrated the outstanding changes in the inner time structures of generated interspike intervals. The output time series of ISIs were studied using non-standard kind of time analysis -- recurrence plot method. Using this technique we observed phenomenon of unification in inner time structures of output series. Further we identified such parameters of model which exerted the most considerable influence on this phenomenon. PMID- 9730058 TI - Effects of new machinable ceramic on behavior of rat bone cells cultured in vitro. AB - The behavior of cultured rat bone cells growing on modified polyethylene terephthalate (mPET), glass, and machinable ceramic substrates containing enstatite (MgO, SiO2) and glass (CaO-P2O5-Al2O3) was studied. Cell attachment was measured directly on the substrates using an image analysis system. Electron microscopy observations and the MTT test revealed that cells are able to spread and proliferate on the material surface, keeping a healthy ultrastructure on all materials tested in the present study. After having colonized the surface of the materials, as shown by immunocytochemistry, the cells synthesize an osteoid-like matrix composed of osteocalcin, type I collagen, and fibronectin fibrils. The titration of alkaline phosphatase activity showed that the cells grown on the ceramic exhibit a greater osteogenic activity than those grown on controls (glass and mPET). This osteogenic activity results in a mineralization of the extracellular matrix in cultures on ceramic or plastic whereas only few calcium phosphate crystallite traces were revealed by Von Kossa staining on glass. Enstatite constitutes, therefore, an environment compatible with in vitro bone cell life. PMID- 9730059 TI - Weibull models of fracture strengths and fatigue behavior of dental resins in flexure and shear. AB - In estimating lifetimes of dental restorative materials, it is useful to have available data on the fatigue behavior of these materials. Current efforts at estimation include several untested assumptions related to the equivalence of flaw distributions sampled by shear, tensile, and compressive stresses. Environmental influences on material properties are not accounted for, and it is unclear if fatigue limits exist. In this study, the shear and flexural strengths of three resins used as matrices in dental restorative composite materials were characterized by Weibull parameters. It was found that shear strengths were lower than flexural strengths, liquid sorption had a profound effect on characteristic strengths, and the Weibull shape parameter obtained from shear data differed for some materials from that obtained in flexure. In shear and flexural fatigue, a power law relationship applied for up to 250,000 cycles; no fatigue limits were found, and the data thus imply only one flaw population is responsible for failure. Again, liquid sorption adversely affected strength levels in most materials (decreasing shear strengths and flexural strengths by factors of 2-3) and to a greater extent than did the degree of cure or material chemistry. PMID- 9730060 TI - Joint fluid from patients with failed total hip arthroplasty stimulates pit formation by mouse osteoclasts on dentin slices. AB - Periprosthetic bone resorption has been implicated in the failure of total joint arthroplasty. Osteolysis is reported to be associated with bone resorption induced by bone-resorbing cytokines, which are released from macrophages and fibroblasts in periprosthetic tissues after stimulation by wear debris generated in the joint cavity. Recent reports have suggested the concept of the effective joint space, which includes all periprosthetic regions that are accessible to joint fluid and wear debris. In this study, we examined the levels of interleukin 6 (IL-6), soluble IL-6 receptor (sIL-6R), and tartrate-resistant acid phosphatase (TRAP) in joint fluid after failed total hip arthroplasty (THA) with osteolysis and investigated whether the joint fluid could activate osteoclastic bone resorption using unfractionated mouse bone cells cultured on dentin slices. Histochemical analysis showed the presence of more TRAP-positive cells in synovial capsules from failed THA patients when compared with osteoarthritis (OA) patients (controls). The levels of IL-6, sIL-6R, and TRAP in joint fluid from failed THA patients were significantly higher than in OA patients. Mouse osteoclasts cultured on dentin slices with joint fluid from failed THA patients with osteolysis produced a significant increase of pit area, whereas cells cultured with joint fluid from OA patients did not. Interestingly, osteoclastic bone resorption on dentin slices was significantly correlated with TRAP activity in joint fluid (p < 0.0001). These results suggest that joint fluid containing bone-resorbing cytokines is produced by synovial capsules in failed THA patients with osteolysis and may activate osteoclasts around the prosthesis in combination with those produced by interface tissues, thus contributing to periprosthetic bone resorption. PMID- 9730062 TI - Dynamic thermomechanical properties and crystallinity of ultrahigh molecular weight polyethylene tibial inserts. AB - A dynamic thermomechanical analysis method was used to determine the values of the viscoelastic parameters, storage modulus (E') and the tangent of the loss angle (tan delta) of samples taken from the near surface and core regions of ultrahigh molecular weight polyethylene (UHMWPE) bar stock specimens that had been aged in ambient laboratory air for 12 months (the control group) and UHMWPE tibial inserts that had been subjected to the following treatments: gamma sterilization followed by shelf aging in ambient laboratory air for between 56 and 112 months or ethylene oxide (EtO) gas sterilization followed by shelf aging in ambient laboratory air for 73 months. Differential scanning calorimetry was used to determine the melt temperature and percentage crystallinity (%C) of these samples. There were three main findings. First, EtO sterilization produced no statistically significant effect on the values of E' or tan delta (relative to those for the control group), but the values of these parameters were markedly affected by gamma sterilization. Second, for gamma-irradiated samples, E' increased with an increase in the shelf aging time. Third, there was a strong direct association between E' and % C. The various potential uses of this association are described, paying special attention to its likely role in predicting the in vivo wear of UHMWPE tibial inserts. PMID- 9730061 TI - Predictive model for tensile true stress-strain behavior of chemically and mechanically degraded ultrahigh molecular weight polyethylene. AB - The gamma radiation sterilization of ultrahigh molecular weight polyethylene (UHMWPE) components in air generates long-lived free radicals that oxidize slowly over time during shelf storage and after implantation. To investigate the combined effects of chemical and mechanical degradation on the mechanical behavior of UHMWPE, sterilized tensile specimens were immersed in 0.5% hydrogen peroxide solution at 37 degrees C for up to 9 months and concurrently subjected to cyclic stress levels of 0 (control), 0 to 5, and 0 to 10 MPa. After chemical and mechanical preconditioning, specimen density was measured using the density gradient column technique. The true stress-strain behavior was measured up to 0.12 true strain and characterized using a multilinear material model, the parameters of which were found to vary linearly with density and cyclic stress history. The mechanical behavior of as-irradiated and degraded UHMWPE was accurately predicted by an analytical composite beam model of the tensile specimens. The results of this study support the hypothesis that chemical and mechanical degradation affect the true stress-strain behavior of UHMWPE. In the future, the material model data presented in this study will enable more accurate prediction of the stresses and strains in UHMWPE components following gamma sterilization in air and subsequent in vivo degradation. PMID- 9730063 TI - Precoating of ultrahigh molecular weight polyethylene with polymethylmethacrylate: interfacial strength. AB - Fixation of polymeric implants, especially an ultrahigh molecular weight polyethylene (UHMWPE) acetabular cup, to a host bone site has been a challenge since its first conception from the Charnley low friction total hip arthroplasty. Destabilization of the acetabular cup, similar to the well-documented cases of femoral stems, is caused mainly by aseptic loosening; the mobile loosened particles further contribute to the progression of aseptic loosening. Although the obvious fixation problems lie in the bone-bone cement interface, little work has been done to reduce the loosening by improving the acetabular components as a whole in cemented procedures. Most of the grooved outer surface, external fixation devices, and metal backings have been introduced to avoid problematic fixation of the cup to bone cement; nevertheless, the designs themselves to some degree became the source of the loosening problems. One possible way to improve the adhesion of acrylic bone cement to the UHMWPE acetabular cup is precoating the surface with polymethylmethacrylate (PMMA). This study successfully precoated the UHMWPE surface with PMMA, showing good chemical and mechanical stability, and suggests the optimal conditions of variables involved in the newly developed precoating process. The highest interfacial tensile strength was 11.51 +/- 0.65 MPa, which is stronger than those of UHMWPE and metal in metal-backed cups (6.3 MPa) and bone-bone cement (8.5 MPa). Further chemical analysis and mechanical testing are in progress, yet the present result of the mechanical tensile strength test showed that the precoating process for the UHMWPE surface could be a viable means toward stable fixation of the polymeric implants by using PMMA bone cement. PMID- 9730064 TI - Subperiosteal implantation of various RF magnetron sputtered Ca-P coatings in goats. AB - The aim of this study was to obtain more information about the initial biological events around RF magnetron sputtered calcium phosphate (Ca-P) coatings. Therefore, uncoated and coated disks were inserted subperiosteal into the tibia of a goat. The coatings were deposited on commercially pure titanium. The thickness of the coating was 0.1 or 2.0 microm. All the as-sputtered coatings were subjected to an additional heat treatment for 2 h at 500 degrees C. After 1 and 3 weeks of implantation the experimental disks were retrieved and prepared for histological and physicochemical analysis. The histological results demonstrated that the periosteum covered the specimens after both implantation periods. In between the periosteum and implant an acellular layer and a collagen matrix was observed. Energy dispersive spectrometry revealed that the acellular layer consisted of C, Ca, and P ions for the 0.1 microm thick Ca-P coatings. The 2 microm thick Ca-P coatings also showed the presence of sulfate ions in this layer. Only organic material was found on the titanium disks. Further, SEM showed that even after 3-week implantation, a substantial thickness of both coatings was still maintained. Thin film X-ray diffraction demonstrated that after both implantation periods, the CaP-0.1 coating was still present. FTIR of the retrieved specimens demonstrated on the coated disks the formation of additional carbonate apatite (CO3-AP) associated with an organic phase (NH2 groups). On basis of these findings we conclude that our experimental approach is very suitable for the investigation of the healing process around Ca-P coatings. Further, we again demonstrated that the initial interfacial response to Ca-P materials differs from titanium. PMID- 9730065 TI - Multilaminate resorbable biomedical device under biaxial loading. AB - The design and test of a multilaminate sheet developed for a hernia repair application is presented. As biomaterial applications become more complex, characterization of uniaxial properties becomes insufficient and biaxial testing becomes necessary. A measure of the in-plane biaxial strength of the device is inferred from a ball burst test. The results of this test for different thicknesses of the device are correlated with the uniaxial strength of the material. A biaxial test such as the ball burst test is more indicative of the properties of a planar material than would be a uniaxial test. The interactions in the biaxial mode of failure are of value and can be related back to a classical uniaxial tensile test from the ball burst test. The material used in this study to fabricate the device was a resorbable biomaterial called small intestinal submucosa (SIS). The effects of rehydration on the stiffness and associated ball burst properties of the SIS device were also measured. It is shown that at a rehydration time of 5 min from a reference dry state, steady state mechanical properties are reached. PMID- 9730066 TI - Gamma irradiation of chitosan. AB - Chitosan has potential biomedical applications that may require the final products to be sterilized before use. The gamma irradiation of purified and highly deacetylated chitosan fibers and films at sterilizing doses (up to 25 kGy) caused main chain scissions. The viscosity average molecular weight of the polymer decreased with increasing irradiation dose, the radiation yields of scission being 1.16 in air and 1.53 in anoxia. Preirradiation application of a negative pressure of 100 kPa disrupted the network structure, which may have contributed to the greater radiation yield obtained by chitosan fibers in anoxia. Radiation induced scission of the chitosan chains resulted in a lower glass transition temperature (Tg), indicative of higher segmental mobility. The Tg was below ambient at an irradiation dose of 25 kGy in air. Irradiation in air improved the tensile strength of the chitosan film, probably due to changes in chain interaction and rearrangement. Irradiation in anoxia did not affect film properties significantly, partly because the preirradiation application of negative pressure had a negligible effect on the structure of the chitosan film. Polymer network structure and the irradiation conditions are therefore important determinants of the extent of radiation induced reactions in chitosan. PMID- 9730068 TI - Analysis of titanium dental implants after failure of osseointegration: combined histological, electron microscopy, and X-ray photoelectron spectroscopy approach. AB - A multitechnique approach has been used to characterize the surface of nonosseointegrated titanium implants and the surrounding biological tissues. Five pure titanium dental implants were used as reference, and 25 removed implants were studied. Surface and in-depth chemical compositions of the implants (from a total of 16 patients) were investigated by X-ray photoelectron spectroscopy (XPS). Histological slides of the surrounding tissues were examined by light microscopy, XPS, and electron microprobe analysis. None of the failed implants presented the regular surface composition and depth profile of the TiO2 overlayer; foreign elements (Ca, Na, P, Si, Cl, Zn, Pb, and Al) were observed on some implants. Fibrosis, lymphocytic and plasmocytic infiltrates, and granulomatous lesions were detected in the surrounding tissues. XPS and electron microprobe analysis indicated the presence of Zn, Fe, Sn, and Ti in the tissues. As a possible scenario for implant failure, we propose and discuss a oxidoreduction mechanism, leading to a partial dissolution or the complete dissociation of the protective titanium dioxide overlayer and to ion diffusion through the surrounding tissues. PMID- 9730067 TI - Analysis of 3-D microstructure of porous poly(lactide-glycolide) matrices using confocal microscopy. AB - Porous matrices of biodegradable polymers are extensively used as scaffolds in tissue engineering and as drug delivery devices. A critical component of the design, processing, and utility of such polymeric systems concerns the local void microarchitecture. In this study, a novel approach based on confocal fluorescence imaging was employed to visualize and quantify in 3 dimensions (3-D) the individual and population-level void morphology within porous polymeric matrices. Poly(lactic acid-glycolic acid) copolymer matrices were cast to yield void configurations of variable void sizes but constant cumulative voidage. Using confocal microscopy, fluorescently saturated polymer matrices were optically sectioned into serial 2-D images, and 3-D void contours were reconstructed via object discrimination and connectivity analysis. The resultant data was used to quantitate the matrix microstructure and map its evolution following polymer degradation. Under conditions of accelerated degradation, matrix erosion was found to cause a significant change in the disposition of voids; this involves two processes (void formation and void enlargement), the extent of which was influenced by the initial void size and the duration of erosion. By virtue of providing both static and dynamic descriptions of the void morphology in poly(lactic acid-glycolic acid) matrices, this is the first spatiotemporal study of the 3-D microarchitecture of porous, bioerodible tissue analog matrices. PMID- 9730069 TI - Oral mucosal adhesive film containing local anesthetics: in vitro and clinical evaluation. AB - In vitro and in vivo studies were conducted to gauge the effectiveness of a novel oral mucosa adhesive, moderately water-soluble, pliant polymer artificial dentifrice (AD) film containing dibucaine (DC) for relief of pain due to oral erosion. The film was prepared from a hydroxypropyl cellulose-M (HPC-M) ethanol solution containing varying amounts of DC, as well as polyethylene glycol. In the in vitro experiments, the disintegration of HPC-M showed a lag time of about 50 min, a much lower rate than that of drug release, which more or less leveled off after 50 min. Twenty-five percent of the DC was released from the film (0.113 and 0.225 mg/cm2) after the initial 5 min, which then reached about 80% after 50 min, the time at which the polymer began to break up. In the in vivo study, the local anesthetic effect of the film was evaluated in 23 patients (10 males, 13 females) suffering from the adverse effects of chemotherapy. When applied to the wet surface of the mucosa, the AD film absorbed moisture and showed excellent adhesion. Pain relief in patients lasted 2.2 +/- 0.21 and 4.3 +/- 0.25 h at DC doses of 0.113 and 0.225 mg/cm2, respectively. These results suggest that the AD film may cover mucositis sites of oral mucosa long enough to allow DC release and bring relief from pain arising from chemotherapy and/or radiotherapy. PMID- 9730070 TI - Reassessment of long-term use of dense HA as dental implant: case report. AB - Dense hydroxyapatite (HA) is widely believed to be unsuitable for clinical use as dental implants due to its poor mechanical properties, although it has excellent biocompatibility and is chemically stable and nonresorbable in vivo. However, the case in this article is one in which the patient's dense HA implants are still stable and in good functional condition 16.5 years after he received four pieces of a one-piece dense HA implant in both sides of his lower molar regions. Furthermore, almost no radiolucency is evident along the root portions of the implant sites in the bone. These findings imply that dense HA can be clinically useful and should be reevaluated as a dental implant material. PMID- 9730071 TI - XPS and SEM detection of surface changes on 64 ureteral stents after human usage. AB - Ureteral stents are commonly implanted to assist the postsurgery flow of urine from the kidney to the bladder. We hypothesized that different surface compositions of stent material could result in different conditioning film depositions and potentially altered receptivity to bacterial biofilms. Using XPS, three types of ureteral stents recovered from 64 patients were found to have adsorbed conditioning films that altered the surface composition of the devices. Elements associated with encrustation (calcium, magnesium and phosphorus) were found on 69% of the silicone latex stents, 44% of the low surface energy (LSe) devices, and 38% of the carbon-rich stents. No statistical difference was found in relation to patient gender, stent type, duration of implantation, and encrustation deposition. The composition of the film suggested that the nature of the underlying material did not significantly alter the elements that adsorbed. Thus, devices may take on a similar surface coat within days, and perhaps hours, of exposure to the host. With respect to dense encrustations, fewer appeared on the LSe devices. SEM confirmed the presence and nature of the film crystals and showed bacterial biofilms adherent to devices and encrustations in three patients who had received prophylactic trimethoprim compared to one on ciprofloxacin. In conclusion, although encrustation deposition and biofilm formation on ureteral stents is not unique to Cook devices, the human model and surface science test systems described here are invaluable to evaluate biomaterials used in patients. Unless biomaterials undergo rigorous analysis in vivo, including true assessment of the outcome of prophylactic antibiotic usage, clinicians will be unable to accurately select the best device and management strategy for a given patient. PMID- 9730072 TI - Ease of donning commercially available latex examination gloves. AB - There are a wide variety of latex examination gloves now available for use by health care providers. A prospective randomized trial was completed to quantify the forces required to don a sample of seven cornstarch-lubricated gloves and 13 powder-free latex examination gloves. The data collected was analyzed by a 20 x 2 general factorial ANOVA, as well as two 1-way ANOVAs using a least significance difference post hoc test. Some powder-free gloves can be easily donned with dry or wet hands without tearing with forces comparable to those encountered with powdered gloves. With the advent of these powder-free examination gloves, powdered gloves can now be abandoned, protecting health professionals and patients from the dangers of absorbable dusting powders. Despite the dangers of the absorbable dusting powders and the Food and Drug Administration's requirement for labeling examination glove boxes, some manufacturers of powdered examination gloves do not appropriately label their boxes with a warning to the health professional and patient of the presence of powder. PMID- 9730073 TI - Concise review of mechanisms of bacterial adhesion to biomaterial surfaces. AB - This article reviews the mechanisms of bacterial adhesion to biomaterial surfaces and the factors affecting the adhesion. The process of bacterial adhesion includes an initial physicochemical interaction phase (phase one) and a late molecular and cellular interaction phase (phase two), which is a complicated process affected by many factors, including the characteristics of the bacteria themselves, the target material surface, and the environmental factors, such as the presence of serum proteins or bactericidal substances. PMID- 9730074 TI - Pediatric psychopharmacology: problems and prospects. PMID- 9730075 TI - Prevalence variations in psychotropic treatment of children. AB - This study was undertaken to clarify several aspects of the estimation of prevalence of three commonly use pediatric psychotropic agents, namely, methylphenidate, desipramine, and imipramine. The study aims are threefold: (1) to show the variability of drug prevalence by comparing estimates from three data sources; (2) to show the misleading impression that can be created by reporting drug prevalence estimates based on counts of prescriptions rather than persons; (3) to show the utility of gender-by-age-specific prevalence of drug use as a marker for diagnosis. Two data sources that yield population-based prescription estimates were available: 1991 Medicaid administrative claims data for prescriptions from a mid-Atlantic state and 1991 prescription records of the northwest region of Kaiser Permanente, a staff-model health maintenance organization (HMO). Another source of data consists of the 1991 National Ambulatory Medical Care Survey, which records medication information reported during physician office visits. Data analysis consists of quantitative estimates of (1) drug prevalence from each source; (2) the ratio of prescription claims to persons; and (3) the proportion of drug use according to age and gender. Methylphenidate and desipramine prevalence had a twofold greater use among state Medicaid enrollees compared with HMO enrollees. Average claims-to-person ratios of 5:1 suggest better accuracy using persons with medication rather than prescription counts. Gender-by-age-specific prevalence rates showed that 75% of the drug use for desipramine among those less than 15 years old was found among males, whereas 75% of the desipramine use among those 15 or older was found among females, suggesting its use for the treatment of attention deficit-hyperactivity disorder among young males and for depression among older females. The variability of community physician decision making in pediatric psychopharmacology is better understood by observing drug prevalence rates from different settings. National sampling efforts should be undertaken to verify regional and setting-specific prevalence findings and to learn the reasons for their fluctuation. PMID- 9730077 TI - Pediatric psychopharmacology in a capitated managed care system: how do patients fare? AB - A retrospective chart review was designed to evaluate how pediatric and adolescent patients fare within a strictly managed, capitated outpatient psychiatric setting. The charts of 96 consecutively evaluated patients, treated by one psychiatrist, were reviewed. Standardized assessment tools were used to reach a DSM-IV diagnosis. The initial level of severity was defined by a Global Assessment of Functioning scale at the time of the evaluation, and improvement after pharmacologic treatment was measured by the Global Index of Improvement. Patients in this sample suffered from similar psychiatric disorders and similar levels of initial impairment when compared with those reported in other outpatient samples. Of the original 96 youths, nine patients were initially referred to their primary care physician for medication treatment, and 26 had been told that pharmacotherapy was not indicated. In the psychiatric clinic, psychopharmacotherapy was recommended for 61 of the 96 patients (64%), and 46 patients (48%) followed through with this plan. Of the 46 patients treated with medications, 72% improved within approximately 4 months of treatment, whereas 28% did not. Five of the 23 (22%) adolescent patients with major depression were hospitalized within 2 months of the initial evaluation. Most of these pediatric and adolescent psychiatric patients appeared to be treated satisfactorily, despite the strong limitations of a capitated outpatient service. However, significant limitations in time and resources necessitated a variety of clinical adaptations to maintain adequate delivery of mental health services. PMID- 9730076 TI - Open-label olanzapine treatment in five preadolescent children. AB - Olanzapine is a recently introduced atypical neuroleptic agent for which little information is available on its use in children. Open clinical trials of olanzapine treatment were conducted on five hospitalized children (ages 6 to 11 years) with varying diagnoses including bipolar disorder, psychosis not otherwise specified, schizophrenia, and attention-deficit/ hyperactivity disorder. Each patient had failed previous psychotropic medication trials, with a mean of four prior trials. The mean length of olanzapine treatment was 32 days (range, 2 to 7 weeks), and mean daily dose was 7.5 mg/day (range, 2.5 to 1.0 mg/day) or 0.22 mg/kg/day (range, 0.12 to 0.29 mg/kg/day). All children experienced adverse effects, including sedation (N = 3), weight gain of up to 16 pounds (N = 3), and akathisia (N = 2). Three patients showed some clinical improvement, but olanzapine treatment was discontinued in all five children within the first 6 weeks of treatment because of adverse effects or lack of clinically significant therapeutic response, although higher (or lower) doses, slower titration of dosage, or a longer duration of treatment might have produced more favorable results. Psychotic symptoms did not respond in the two patients with evidence of overt hallucinations and paranoid ideation. Improvement was observed in sleep in all five patients and in control of aggression in three. Before controlled trials of olanzapine in children are undertaken, further exploration of dose range and increased duration of treatment on an open basis are warranted. Until more encouraging data are available, clinicians should be cautious and conservative in their predictions about the potential value of olanzapine in treating preadolescent psychiatric disorders. PMID- 9730078 TI - Prediction of stimulant response in children with attention-deficit/hyperactivity disorder. AB - Discrimination of stimulant-responding and nonresponding groups of children with attention-deficit/hyperactivity disorder (ADHD) on the basis of demographic, neurophysiologic, or behavioral variables would be beneficial for clinical and theoretical reasons. Previous researchers have identified many predictor variables, but relationships between predictor and criterion variables generally have been subtle. In addition, few investigations have considered the relative predictive power of the variables. The present study evaluated the multivariate relationship between several predictor variables and response to medication in 336 children with ADHD. Neurologic status, inattention, and overactivity were found to be most likely to predict good response to psychostimulants, whether rated by parents or teachers. Although a number of variables predicted a positive psychostimulant response, the strength of the predictive associations suggests only minimal clinical usefulness. PMID- 9730079 TI - Predictors of medication compliance after hospital discharge in adolescent psychiatric patients. AB - Failure in medication compliance in adult psychiatric patients is often found to be due to side effects or associated with unawareness of illness. Little research has been conducted on medication compliance in adolescent psychiatric patients. In this study, 97 adolescent psychiatric patients, including 46 with substance abuse, were followed up an average of 14 months after their discharge from inpatient psychiatric care. Compliance with prescribed medications was examined and the association between several potential predictors and compliance was examined. The overall rate of medication compliance was only 38% at 14-month follow-up, whereas the rate of patients stopping their medications because of side effects was only 23%. Significant predictors of compliance failures were general noncompliance with the discharge plan, followed by postdischarge substance abuse. Side effects did not contribute any additional variance when these factors were considered. These data suggest that medication compliance may be adversely impacted by general tendencies toward noncompliance with treatment, which may be mediated by several possible factors. Interventions to increase awareness of the need for psychotropic medications as well as careful monitoring for substance abuse relapse in this population are suggested. PMID- 9730080 TI - Increased plasma valproate concentrations when coadministered with guanfacine. AB - A preliminary clinical observation suggests the possibility of changes in valproate level when coadministered with guanfacine. Two pediatric inpatients (8 and 9 years of age) were treated with valproic acid and guanfacine concurrently. In one child, when guanfacine was tapered and discontinued, the plasma valproate concentration decreased by 41% from 128 microg/mL to 76 microg/mL. In the other case, studied in an ABA design, the child exhibited a rapid increase in plasma valproate levels while guanfacine was administered. It is proposed that guanfacine may affect the pharmacokinetics of valproic acid and lead to a significant increase in valproate plasma levels when used concurrently with this agent. The mechanism of this proposed interaction may involve drug-drug competition at the level of hepatic glucuronidation (conjugation), although shifts in protein binding cannot be ruled out. PMID- 9730081 TI - Does guanfacine trigger mania in children? PMID- 9730082 TI - Immunoassay of human Neisseria meningitidis serogroup A antibody. AB - An enzyme immunoassay is described which quantitates antibodies to Neisseria meningitidis serogroup A capsular polysaccharide in human sera. Modifications of a previously developed two-day assay by Carlone et al. were made to permit analysis in one day and to be compatible with automation. The allowable variations in assay conditions and the areas in which stringent control must be maintained for consistent assay performance are described. Antigen-coating parameters, the kinetics of primary and secondary antibody incubation steps, the buffer compositions, including detergents, serum requirements, and the need for blocking steps were examined. Our modified one-day assay showed excellent agreement with the standardized method of Carlone, with a correlation coefficient between the two methods of 0.989. This assay is adaptable within a permissible range of parameters thus facilitating the implementation of the standardized assay. This will maximize the consistency of results from serum analysis for conjugate vaccine trials related to serotype A Neisseria meningitidis. PMID- 9730083 TI - The non-specific binding of immunoglobulins to silicone implant materials: the lack of a detectable silicone specific antibody. AB - Recent studies have suggested that anti-silicone antibodies develop in patients implanted with silicone materials. The majority of these studies have utilized enzyme-linked immunosorbent assay (ELISA) methodology with a silicone material substrate as a means to detect the presence of the anti-silicone antibody. The current studies were undertaken to determine whether the binding of IgG to a silicone substrate was consistent with an antigen-specific antibody interaction or the result of non-specific hydrophobic interactions. While significant differences were detected in serum from silicone antibody "positive" and "negative" patients when the ELISA was conducted using a phosphate buffered saline (PBS)-0.05% Tween 20 (Tween) blocking system, the difference in the responses was attenuated when protein blocking systems were used or when incubation times were decreased. Furthermore, ELISA studies, using purified mouse and human IgG, demonstrated a concentration-dependent binding of IgG to silicone elastomer substrate which was also attenuated when a protein blocking system was used in lieu of Tween. In controlled animals studies in which female B6C3F1 mice were implanted with silicone gel or silicone elastomer for 180 days, no difference was observed between the implanted animals and the PBS control animals with respect to binding of IgG to the silicone substrate. Similar studies in female Fischer 344 rats implanted with silicone gel for 84 days also failed to demonstrate the presence of anti-silicone antibody. Collectively, the results suggest that the binding of IgG to silicone implant materials is non-specific in nature, consistent with the well-recognized interactions between hydrophobic molecules (IgGs) and hydrophobic surfaces (silicones) in an aqueous-based system. PMID- 9730084 TI - T lymphocyte subsets and activation status in patients following liver transplantation. AB - Changes in T-lymphocyte subsets have previously been shown to relate to clinical events following liver transplantation and be of prognostic significance following renal transplantation. The aim of this study was to examine T lymphocyte subsets, their activation status and the mean fluorescence intensity of cell surface markers by flow cytometric analysis, in peripheral blood of patients following liver transplantation. Stable transplant patients (n=11) had a significantly higher level of activation (HLA-DR expression ) of all T cell subsets: CD3, CD4 and CD8 compared to healthy controls: 17.5% +/- 14.0 (mean +/- SD) vs 4.7 +/- 1.8 (p=0.04), 13.7% +/- 10.3 vs 4.3 +/- 1.7 (p=0.03) and 23.8% +/- 19.9 vs 3.6 +/- 2.4 (p=0.02) respectively. A further increase in activation status occurred in all T cell subsets in association with acute cellular rejection, reaching significance for the CD4+ population: 13.7% +/- 10.2 vs 23.3% +/- 20.6 (p=0.04). The mean fluorescence intensity of the CD3+DR- and CD3+ DR+ populations were increased to 1397 +/- 869 and 1282 +/- 810 following liver transplantation compared to values of 425 +/- 204 and 376 +/- 166 respectively for controls (p<0.05). T-lymphocytes maintain a high level of activation following liver transplantation and continue to express high levels of the surface marker CD3, which may account for the occurrence of acute cellular rejection despite immunosuppression in these patients. PMID- 9730085 TI - Late expression of tumor necrosis factor-alpha is markedly depressed in patients with IgA nephropathy. AB - In this study we demonstrate that peripheral blood mononuclear cells (PBMC) of patients with IgA nephropathy (IgAN) express mRNA for tumor necrosis factor alpha (TNF-alpha) at a significantly lower frequency after stimulation for 24 hours with pokeweed mitogen than do PBMC of a control population. These differences were not seen in patient samples stimulated for only 3 hours. To identify the cells responsible for TNF-alpha expression after overnight mitogen stimulation, we performed reverse transcriptase polymerase chain reaction analysis after preparative flow cytometry of PBMC labelled with fluorescent monoclonal antibodies. After stimulation for 24 hours, PBMC of all patients tested displayed no detectable TNF-alpha mRNA; in the control PBMC, TNF-alpha mRNA was present in CD4+ and CD8+ T cells, as well as in CD19+ B cells in some samples. These results suggest that diminished expression of TNF-alpha by T cells may play a role in the pathogenesis of IgAN. PMID- 9730086 TI - Specificity of monoclonal antibodies to Campylobacter jejuni lipopolysaccharide antigens. AB - Monoclonal antibodies (Mabs) were produced to the lipopolysaccharide antigens of Campylobacter jejuni strain 1249 (Penner serotype O:2/63). A polymyxin-cloth based enzyme immunoassay (pCEIA) was used for initial screening and for evaluating the specificity of these antibodies. Seven Mabs reacted with at least 11 and as many as 14 of 15 C. jejuni strains (representing 8 different Penner serotypes). These seven Mabs did not cross-react with any of 16 non-Campylobacter bacteria commonly encountered in food, with only two exceptions. Several combinations of these Mabs in pairs reacted with all 15 C. jejuni strains. These results suggest that pCEIA employing two of these Mabs in combination is potentially useful for detection of Campylobacter jejuni in foods and other samples. PMID- 9730088 TI - TAP1 polymorphism identified in African-American Graves' disease patients. AB - Polymorphism in transporter associated with antigen processing (TAP)1 gene has been observed in African American Graves' disease patients. Single strand conformational polymorphism has been used to identify variation for the locus. First-strand cDNA was generated from cell lines obtained by Epstein-Barr virus immortalization. Four variant alleles for TAP1 have been observed and the products have been sequenced to compare with the location of observed with SSCP position patterns. Variants were detected and compared with substitutions within TAP1 polypeptide which includes changing valine to leucine and three (3) silent substitutions for glycine, glutamic acid and alanine. PMID- 9730089 TI - Exogenous dehydroepiandrosterone modified the expression of T helper-related cytokines in NZB/NZW F1 mice. AB - The onset of lupus-like disease in NZB/NZW F1 mice was correlated with the expression of IL-10 at 4 m of age, and with a sequential enhanced expression of IFN-gamma and IL-6 between 6 to 8 m of age. The expression of IFN-gamma and IL-6 was associated with exacerbation of disease symptom, production of anti-DNA antibody, and increase in total serum IgG1. Exogenous dehydroepiandrosterone (DHEA) given in animal diet significantly prolonged survival, and delayed formation of autoantibody of NZB/NZW F1 mice as compared to mice fed on control diet. The effect of DHEA paralleled a delay in the expression of IL-10 and IL-6 and an earlier detection of IL-12 transcripts. Moreover, DHEA-fed mice had higher serum IgG2a level than control diet-fed mice. Collectively, DHEA may modify the activation of distinct subset of T helper cells in NZB/NZW F1 mice at different phases of disease progression. PMID- 9730087 TI - Interaction between interleukin 10 and interleukin 6 in human B-cell differentiation. AB - Contrary to their opposing action on human T-lymphocytes and monocytes, both Interleukin (IL-)10 and IL-6 are potent differentiation factors of human B-cells. Both are known to induce immunoglobulin (Ig) production. The precise mechanism of this converging effect of IL-6 and IL-10 remains elusive, however. Here we investigated the role of IL-6 in the IL-10 dependent B-cell differentiation into Ig secreting cells. We found that co-stimulation of SAC-stimulated human peripheral B-lymphocytes with IL-10 and IL-6 exhibited no additive effect on Ig production over stimulation with IL-10 alone, and that IL-6 receptor blockade only mildly inhibited IL-10 induced Ig synthesis. In fact, we could show that stimulation of B-cells with IL-10 somewhat suppressed SAC induced autocrine IL-6 production. Despite this suppression IL-6 levels remained sufficiently high to stimulate its receptor, and IL-6 binding to the B-cell surface was not affected. The failure of IL-6 to exert an additional effect on SAC+IL-10 induced Ig production suggests that IL-10 may recruit components of the IL-6 intracellular pathway for Ig induction. In conclusion we could demonstrate that IL-10 acts on B cell differentiation independently of autocrine IL-6 and that it had a considerably mild effect on B lymphocytic autocrine IL-6 secretion. This still allows an IL-6 effect in the presence of IL-10 which appears adaptive with a view to the converging effects of these two cytokines on human B lymphocytes. Our study thus adds to the appreciation of the complex cytokine regulation of the immune system. PMID- 9730090 TI - T cell receptor V-segment frequencies in peripheral CD8+ T cells in aging. AB - Infections are a major cause of illness and death amongst elderly people. Peripheral blood CD8+ T lymphocytes --which play a crucial role in host defence against viral infections--, are divided in subsets based upon the expression of several cell and activation markers. In senescence changes and variations in peripheral CD8+ T lymphocyte compartment have been described, and such a decrease in the CD8+CD45RA+ lymphocytes. In this report the T V alpha and Vbeta cell specificities repertoire usage in aging subjects were studied by the use of seven different monoclonal antibodies specific for defined regions of the TCR. Except for the Vbeta6subfamily, no differences between old and control subjects in frequency of T cells bearing selected V alphabeta epitopes, were observed. PMID- 9730092 TI - Outcome research: definitions and directions. PMID- 9729611 TI - Linkage map of Escherichia coli K-12, edition 10: the traditional map. AB - This map is an update of the edition 9 map by Berlyn et al. (M. K. B. Berlyn, K. B. Low, and K. E. Rudd, p. 1715-1902, in F. C. Neidhardt et al., ed., Escherichia coli and Salmonella: cellular and molecular biology, 2nd ed., vol. 2, 1996). It uses coordinates established by the completed sequence, expressed as 100 minutes for the entire circular map, and adds new genes discovered and established since 1996 and eliminates those shown to correspond to other known genes. The latter are included as synonyms. An alphabetical list of genes showing map location, synonyms, the protein or RNA product of the gene, phenotypes of mutants, and reference citations is provided. In addition to genes known to correspond to gene sequences, other genes, often older, that are described by phenotype and older mapping techniques and that have not been correlated with sequences are included. PMID- 9730091 TI - Functional significance of adhesion molecules in Fas-dependent apoptotic cell death induced by interleukin-2-activated T cells. AB - We investigated the functional significance of the adhesion molecules CD2 and lymphocyte function-associated antigen-1 (LFA-1: CD11a/CD18) in Fas-Fas ligand (FasL) death pathway. Interleukin-2-activated T cells expressed a large amount of FasL protein and could efficiently kill a Fas-sensitive leukemic cell line, MML 1. The major part (over 80%) of MML-1 cell death was Fas-dependent. Antibodies to CD2 and CD11a/CD18 completely inhibited MML-1 target cell lysis, whereas effector to target cell binding was partially reduced or not affected at all. These results suggest that effector/target interaction via CD2/CD58 and LFA-1/CD54 systems would be essential for triggering target cell death. More interestingly, there is the discordance in the ability of anti-CD2, and particularly anti-LFA-1 antibodies, to block Fas-dependent cell death versus effector to target conjugate formation. This suggests some non-adhesive role for CD2 and LFA-1 in induction of Fas-dependent cell death. Although these antibodies were capable of inhibiting T cell proliferative response, there was no significant reduction of FasL or granzyme B expression. Thus, the signaling pathway for growth inhibition via CD2 and LFA-1 could not be linked to signaling for FasL and granzyme B expression. PMID- 9730093 TI - Nerve and tendon gliding exercises and the conservative management of carpal tunnel syndrome. AB - While developments continue in the surgical management of carpal tunnel syndrome, little emphasis has been placed on the evaluation of a comprehensive non-surgical treatment. In this study, 197 patients (240 hands) presenting for treatment of carpal tunnel syndrome were divided into two groups. Patients in both groups were treated by standard conservative methods, and those in one group were also treated with a program of nerve and tendon gliding exercises. Of those who did not perform the nerve and tendon gliding exercises, 71.2% underwent surgery compared with only 43.0% of patients who did perform them. Patients in the experimental group who did not undergo surgery were interviewed at an average follow-up time of 23 months (range, 14-38 months). Of these 53 patients, 47 (89%) responded to this detailed interview. Of the 47 who responded, 70.2% reported good or excellent results, 19.2% remained symptomatic, and 10.6% were non compliant. Thus, a significant number of patients who would otherwise have undergone surgery for failure of traditional conservative treatment were spared the surgical morbidity of a carpal tunnel release (p = 0.0001). PMID- 9730094 TI - The effects of arm elevation and overnight head-up tilt on forearm and hand volume. AB - Methods to reduce increased fluid volume, or swelling, were evaluated as short- and long-term interventions. Forearm and hand volumes were measured in 45 fit and healthy subjects using a water displacement device with previously established reliability. Volumes were measured before and after 2 hours of recumbency and before and after overnight sleep under different conditions of arm elevation or head-up tilt. No arms-at-side lying-down position, whether after 2 hours awake or after overnight sleep or with bed-head elevated or not, resulted in significant changes in forearm and hand volume. Only 2 hours in a supine lying-down position with 30 degrees of arm elevation caused a significant effect, with an average decrease of 51 ml in forearm and hand volume. PMID- 9730101 TI - Noblesse oblige. PMID- 9730097 TI - An alternative splint design for trigger finger. AB - Conventional resting splints used to treat digital stenosing tenosynovitis (trigger finger) are often discontinued or not worn consistently by patients. Informal reports by patients indicate that the splints are too bulky, interfere with activities of daily living, and are visibly too noticeable. Since resolution of the condition may take as long as 9 weeks, this is a significant issue. A new splint design has been developed to avoid these shortcomings in hopes that physicians and therapists will have successful compliance when utilizing splinting in the treatment of this condition. It is both cost-effective and noninvasive. This article describes the condition and treatment options for digital stenosing tenosynovitis and fabrication techniques for the proposed alternative splint. PMID- 9730098 TI - Bio-dynamic finger component. PMID- 9730099 TI - Evaluating research when "no significant differences were found": the issue of statistical power. PMID- 9730103 TI - Leg blood flow and increased potassium release during exercise in chronic heart failure: effect of physical training. AB - BACKGROUND: Exercise-induced hyperkalemia, which may contribute to exercise hyperpnea and exertional fatigue, is increased in patients with chronic heart failure (CHF). This study examined whether differences in leg blood flow during exercise could be responsible for alterations in the level of hyperkalaemia, as well as the effect of physical training. METHODS AND RESULTS: We studied 10 subjects with CHF (ejection fraction 23 +/- 3.9%; mean +/- SD) and 10 subjects with normal left ventricular function (NLVF) who had undergone previous coronary bypass graft surgery (ejection fraction 64 +/- 8.0%; mean +/- SD). Subjects performed incremental cycle exercise to exhaustion before and after physical training. The rises in femoral venous potassium concentration ([K+]), heart rate, lactate, and ventilation (VI) with exercise were all greater in the subjects with CHF than in those with NLVF (P < .05). There was no difference between the groups in leg blood flow during submaximal exercise but peak leg flow was greater in the group with NLVF (P < .01). Physical training was well tolerated and both groups increased their peak VO2 (8 +/- 3.2% CHF (P < .05); 11 +/- 2.7% NLVF (P < .01); mean +/- SE). Training resulted in a reduced rise in femoral venous [K+] and VI (P < .05), but did not affect leg blood flow during submaximal exercise in either group. CONCLUSIONS: The rise in the femoral venous [K+] with exercise is increased in patients with CHF and can be reduced by physical training. These changes are not a consequence of different leg blood flows, either between groups or with training. The study also suggests that femoral venous [K+] is not a powerful regulator of leg blood flow during exercise. PMID- 9730102 TI - Muscular performance in heart failure. AB - BACKGROUND: Some of the major symptoms in patients with chronic heart failure are muscle weakness and fatigue. However, not much is known about muscle performance in these patients compared to healthy controls. METHODS AND RESULTS: Activity level, gait speed, hand grip strength, muscle performance of the knee extensors and flexors along with the plantar and dorsal flexors of the foot were evaluated. Muscle biopsies from the lateral vastus lateralis were taken. Sixteen patients in New York Heart Association class II or III were tested and compared to 112 reference subjects. Compared to the reference subjects, there was a reduction in activity level, gait speed, isometric and isokinetic peak torque for knee extension at different velocities, hand grip strength, peak torque for plantar and dorsal flexion of the ankle and isometric and isokinetic endurance for the knee extension. Recovery was faster. There were small differences in fiber composition. 3-Hydroxy-acylCoA-dehydrogenase and citrate synthase were lower, and lactate dehydrogenase was increased. CONCLUSIONS: Muscle performance is affected in terms of both strength and endurance, which might affect performance in everyday activities. The more pronounced reduction in hand grip compared to the other muscles tested could be an indication of intrinsic abnormalities in the skeletal muscle. PMID- 9730096 TI - Normal digit tip values for the Weinstein Enhanced Sensory Test. AB - The Weinstein Enhanced Sensory Test (WEST) is performed using a calibrated monofilament esthesiometer. This study was designed to establish normal values for the WEST when testing digit tip sensibility and to assess whether factors such as age, gender, and side affect normal WEST values. In order to establish normal values, the WEST was performed on 120 subjects who had no clinical evidence of peripheral neuropathy or subjective changes in digit tip sensation. Subjects also completed a questionnaire and underwent neurometric testing of the median nerve and a brief clinical examination. The results of the study indicate that age had a significant effect on WEST values (p = 0.0002) and that there was an interaction effect for age and gender (p = 0.018). There are strong correlations between the WEST values for individual digits for a given subject and between WEST and electroneurometer values. Normal values for the WEST should be interpreted with regard to age and gender. These normal values (defined as the values for 80% of the population of each category) based on the sample used for this study are as follows: for men and women 55 years of age or younger, 0.035 g; for women older than 55 years, 0.15 g; for men older than 55 years, 0.385 g. PMID- 9730095 TI - Presentation and response of patients with upper extremity repetitive use syndrome to a multidisciplinary rehabilitation program: a retrospective review of 24 cases. AB - OBJECTIVE: To analyze retrospectively a group of patients presenting to an outpatient hand rehabilitation clinic with complaints related to repetitive tasks of the upper extremity. DESIGN: Retrospective case study reviewing 24 consecutive cases for presenting symptoms and response of patients to a multidisciplinary rehabilitation approach. SETTING: An outpatient hand rehabilitation clinic in a tertiary referral center offering simultaneous medical, psychological, and occupational evaluations. PATIENTS: Twenty-four patients with upper extremity symptoms related to repetitive use, who had all failed various prior therapeutic interventions. Fifty percent of the patients were receiving medical disability compensation because of their symptoms. Sixty-two percent had filed a worker's compensation claim. INTERVENTIONS: Treatment consisted of medical management with pharmacologic interventions, occupational therapy with workplace simulation and job-site evaluations, and psychological treatment with pain management and biofeedback training. Treatments were individualized to meet each patient's needs. OUTCOME MEASURES: Reduction in symptom intensity or frequency, increase in work and performance of activities of daily living, and termination of medical disability with return to work. RESULTS: Most cases (83%) were found to be related to occupational computer keyboard use. Bilateral hand and forearm pain were the major symptoms. A unique physical finding was diffuse tendon tenderness and tightness of the long flexor and extensor muscles of the forearm. Carpal tunnel syndrome was found in only one patient. Twenty-five percent of patients achieved resolution of most symptoms, although on a modified and often reduced activity level; 54% had moderate improvement; and 13% had only minimal or no improvement. Of the patients receiving medical disability compensation, 58% returned to their previous jobs. CONCLUSIONS: Patients with upper extremity symptoms related to repetitive use often have unique physical findings, distinct from those of carpal tunnel syndrome. Resulting work disability is high. Patients who have not responded to conventional interventions within a reasonable time may benefit from a multidisciplinary treatment approach. Most patients improve with this treatment but do not fully recover. PMID- 9730104 TI - Enalapril restores depressed circulating insulin-like growth factor 1 in patients with chronic heart failure. AB - BACKGROUND: Congestive heart failure (CHF) is characterized by increased activity of the renin-angiotensin system. Recent experimental studies have shown that infusion of angiotensin II results in depressed plasma levels of insulin-like growth factor 1 (IGF-1) and weight loss. We have previously reported that stable patients with CHF have decreased activity of the growth hormone (GH)-IGF1 axis. We have hypothesized, therefore, that angiotensin-converting enzyme (ACE) inhibition therapy should restore GH-IGF1 activity in CHF patients. METHODS AND RESULTS: Nine patients with stable CHF who were taking digitalis and diuretics, New York Heart Association functional class III were studied before and after 8 weeks of therapy with Enalapril (10 mg twice daily). We measured IGF1 levels, radionuclide left ventricular ejection fraction (EF) and peak oxygen consumption (PVO2). We found that 7 of 9 patients had abnormally low levels of IGF1 (0.2-0.5 mU/ml). IGF1 levels reverted to normal after Enalapril therapy (0.36 +/- 0.03 to 0.8 +/- 0.14 mU/ml, P = .004). This was associated with a significant increase in EF (27.4 +/- 1.1 to 31.4 +/- 0.9%) and PVO2 (14.8 +/- 1.2 to 18.6 +/- 1.5 ml/kg/min) values (P < .05). CONCLUSION: Chronic ACE inhibition therapy restored previously reduced IGF1 plasma levels in patients with CHF, most likely by reducing angiotensin II activity. PMID- 9730106 TI - Should manipulation of myocardial substrate utilization patterns be a component of the congestive heart failure therapeutic paradigm? PMID- 9730105 TI - Beta-receptor blockade decreases carnitine palmitoyl transferase I activity in dogs with heart failure. AB - BACKGROUND: Pharmacological inhibition of carnitine palmitoyl transferase I (CPT I), the enzyme controlling the rate of fatty acid transport into the mitochondria, prevents the contractile dysfunction, myosin isozyme shift and deterioration in sarcoplasmic reticulum Ca2+ handling that occurs in rat models of left ventricular hypertrophy. In this study we examine whether the improved cardiac function with beta blockade therapy in heart failure is associated with an alteration in CPT-I activity. METHODS AND RESULTS: We examined dogs with coronary microembolism-induced heart failure treated for 12 weeks with metoprolol (25 mg twice daily). Myocardial activities of CPT-I, medium-chain acyl co-enzyme A dehydrogenase (MCAD, a beta-oxidation enzyme), citrate synthase, and triglyceride content were measured. The progressive decrease in cardiac function was prevented by treatment with metoprolol, as reflected by an improved ejection fraction over 12 weeks in the metoprolol group (from 35% to 40%) compared to the untreated heart failure dogs (decrease from 36% to 26%). Dogs treated with metoprolol had a marked decrease in CPT-I activity (0.46 +/- 0.03 vs. 0.64 +/- 0.02 micromol min(-1) g(-1) wet weight; P < .02) along with an increase in triglyceride concentration compared to untreated heart failure dogs (3.9 +/- 0.3 v 4.9 +/- 0.2 micromol/g wet weight, respectively; P < .003). By contrast, MCAD and citrate synthase activities did not change. CONCLUSION: Metoprolol induced a decrease in CPT-I activity and an increase in triglyceride content. These results suggest that the improved function observed with beta blockers in heart failure could be due, in part, to a decrease in CPT-I activity and less fatty acid oxidation by the heart. PMID- 9730107 TI - Time-related adaptations in plasma neurohormone levels and hemodynamics after myocardial infarction in the rat. AB - BACKGROUND: Neurohormonal activation plays an important role in the progression of heart failure. In this study, we investigated the progression of neurohormonal activation in conjunction with the hemodynamic status of the rat after myocardial infarction (MI). METHODS AND RESULTS: Male Wistar rats were subjected to sham or MI surgery. At 1, 3, 5, and 13 weeks after surgery, cardiac output (CO), mean arterial pressure (MAP), and total peripheral resistance (TPR), were measured. In separate groups of rats, blood was sampled at 1, 5, and 13 weeks after surgery and analyzed for various neurohormones. At 1 week after surgery, CO and TPR were not altered in MI rats, but plasma neurohormonal levels were elevated. At 3 and 5 weeks after surgery, reduced CO, increased TPR, and normal MAP were measured in MI rats compared to sham rats, but only endothelin levels were elevated. At 13 weeks after surgery, MAP was reduced in MI rats and CO and TPR were comparable between groups. Neurohormonal activation was again apparent in MI rats. CONCLUSIONS: Myocardial infarction in the rat induces early neurohormonal activation, which normalizes hemodynamic parameters. A compensatory phase follows. At 13 weeks after MI, plasma concentrations of neurohormones are again elevated, perhaps as a sign of transition to decompensation. PMID- 9730108 TI - The brain is a possible target for an angiotensin-converting enzyme inhibitor in the treatment of chronic heart failure. AB - BACKGROUND: Although many lines of evidence have shown beneficial effects of angiotensin-converting enzyme (ACE) inhibitors on patients with chronic heart failure (CHF) after myocardial infarction (MI), the target of ACE inhibitors still remains unclear. The objectives of the present study were to evaluate the dipsogenic response to centrally administered angiotensin and to examine the effect of central administration of an ACE inhibitor on cardiac remodeling in rats with CHF after large MI. METHODS AND RESULTS: The drinking responses to intracerebroventricular (i.c.v.) injections of saline and angiotensin I (100 ng) were measured in 22 male Sprague-Dawley rats with or without CHF at 2-5 weeks after the ligation of the left coronary artery. The dipsogenic responses to i.c.v. angiotensin I were significantly larger in rats with CHF and large MI (infarct size > 30%) than in sham-operated rats. Pretreatment with losartan abolished the significant difference between the two groups. Left ventricular (LV) weights of 32 surviving rats with CHF were measured after the 3-week subcutaneous infusions of vehicle (s.c.-VEH) and captopril (1 mg x kg(-1) x h( 1), s.c.-CAP) or the 3-week i.c.v. infusions of vehicle (i.c.v.-VEH) and captopril (50 microg x kg(-1) x h(-1), i.c.v.-CAP). The LV weights normalized by body weights of s.c.-CAP rats were significantly smaller than those of s.c.-VEH rats (1.73 +/- 0.04 vs 2.08 +/- 0.09 g x kg(-1); P < .01); those of i.c.v.-CAP rats were also significantly smaller than those of i.c.v.-VEH rats (1.84 +/- 0.08 vs 2.1 +/- 0.10 g x kg(-1); P < .05). CONCLUSIONS: These results suggest that the brain is a possible target for ACE inhibitors in the treatment of CHF after MI. PMID- 9730109 TI - The use of pacemakers as treatment for systolic dysfunction. PMID- 9730110 TI - Effects of hydration on tactile sensation. AB - The routine tasks of washing usually necessitates the immersion of parts of the body in water, which causes hydration and changes in the mechanical properties of the superficial layer of skin. To determine how hydration affects tactile sensations, the hydration and skin-surface temperature of glabrous and hairy skin was first measured under normal conditions (air), after submersion in distilled water alone and after submersion in a surfactant-water solution. In these experiments, measurements were made of the time to achieve complete hydration and the recovery time to normal levels. The uptake of water in hairy skin was found to be considerably greater than in glabrous skin, and retention was significantly prolonged by the surfactant additive. Subsequent experiments on glabrous skin, based on the results of the preceding hydration studies, measured in-air and hydrated tactile thresholds and sensation magnitudes to vibratory stimuli and to the roughness of textured surfaces. Vibrotactile detection thresholds were not affected by skin hydration, nor were sensation magnitudes to suprathreshold vibratory stimuli. However, suprathreshold perceptions of roughness were substantially altered by hydration. It is concluded that hydration and the mechanics of the skin play a major role in the perception of spatiotemporal (i.e., textured) surfaces and, thus, must be taken into account in any physiological/psychophysical model based on using such stimuli. This may not be required for models based on predominantly temporal (i.e., vibratory) stimuli. PMID- 9730112 TI - Human cerebrocortical potentials evoked by stimulation of the dorsal nerve of the penis. AB - Cortical evoked potentials resulting from stimulation of the dorsal nerve of the penis (DNP) provide a unique opportunity to document the cortical localization of sexual sensory representation in man. The DNP supplies sensory axons to the major portion of the human phallus, including the penile shaft and glans. Animal and human studies indicate that this nerve plays a crucial role in erection and ejaculation. Direct cortical evoked responses to DNP electrical stimulation were recorded in patients undergoing preoperative evaluation for resection of epileptic foci. These studies provided evidence that the primary sensory cortex contains a large area of cortex devoted to the afferent fibers of the DNP and that the sensory field is in a different location than previously described. The location and distribution of this response indicated the need for revision of the traditional concept of the sensory cortical homunculus. PMID- 9730111 TI - Contribution of a Ia muscle afferent activation to the rise of H reflexes and somatosensory evoked potentials in man. AB - Electrical stimuli were applied to the tibial nerve in the popliteal fossa in man in order to investigate how information is transferred from group I muscle afferents to motoneurons and to the somatosensory cortex. For control purposes, identical stimuli were applied to the skin beside the electrode above the nerve. The somatosensory evoked potential (SEP) to skin stimulation alone had a peak latency which was 5 ms longer than the SEP to transcutaneous nerve stimulation. Influences by stimulation of the skin above the nerve could thus be excluded. The threshold intensity to evoke a liminal H reflex was at least two times higher than the threshold for a SEP. In most of the subjects, there was a correlation between the H reflex and the SEP size. If two identical stimuli were applied to the posterior tibial nerve with an interval of 1 s, the second H reflex was 30% smaller than the first one (postactivation depression). The corresponding SEPs were, however, only slightly reduced. Postactivation depression was probably caused by general intrinsic properties of synapses of group I muscle afferents. The results of this investigation indicate that: (1) a large volley in group I muscle afferents is necessary to evoke a liminal H reflex, whereas transmission from muscle afferents to the somatosensory cortex is very efficient; (2) these feedback signals to motoneurons and the somatosensory cortex are used independently. PMID- 9730114 TI - A ratio code for vibrotactile pitch. AB - Subjective impressions of pitch for 80 different sinusoidal vibrotactile stimuli delivered to the index finger were measured by free magnitude estimation in four subjects. In three of the subjects, pitch at a given frequency decreased as stimulus amplitude increased. The data of these subjects were well described by a model of pitch based on the relative levels of activation of the three major tactile channels. The main element in this model was a ratio of P channel activity to the sum of the activity levels of the P, NPI, and NPIII channels. Activity levels of the channels were estimated on the basis of the psychophysical literature, including a study of vibrotactile loudness using the same subjects and stimuli as those employed here. A fourth subject, whose pattern of loudness judgments had previously been shown to differ from those of the other subjects, did not conform to this pitch model: her data revealed significant increases in pitch with increases in amplitude, and appear to reflect an inability to combine signals across vibrotactile channels. Pitch changes resulting from vibrotactile adaptation were directionally consistent with our ratio model: pitch was slightly increased by adaptation to a 25 Hz stimulus, and slightly decreased by 200 Hz adaptation. PMID- 9730113 TI - S100 protein-immunoreactive trigeminal neurons innervating the rat molar tooth pulp. AB - S100-immunoreactivity (ir) was examined in tooth pulp primary neurons of the rat. An immunofluorescence method demonstrated that the molar tooth pulp contained S100-immunoreactive (ir) nerve fibers. In the root pulp, pulp horn and roof of the pulp chamber, S100-ir smooth and varicose fibers ramified and formed subodontoblastic nerve plexuses. All the fibers became varicose at the base of the odontoblastic layer and extended to the odontoblastic layer. Some varicose endings could be traced into the dentin. The trigeminal neurons retrogradely labeled with fluorogold (FG) from the first and second maxillary molar tooth pulps exhibited S100- and parvalbumin-ir. Approximately 60% and 24% of the labeled cells were ir for S100 and parvalbumin, respectively. Virtually all parvalbumin-ir FG-labeled cells showed S100-ir, while 40% of S100-ir ones coexpressed parvalbumin-ir. An immunoelectron microscopic method revealed that all myelinated axons and half of the unmyelinated axons in the root pulp contained S100-ir. In the odontoblastic layer, predentin and dentin, S100-ir neurites lost the Schwann cell ensheathment and made close contact with cell bodies and processes of odontoblasts. The odontoblastic layer also contained parvalbumin-ir neurites. These neurites were devoid of the Schwann cell ensheathment and in close apposition to cell bodies and processes of odontoblasts. S100-ir pulpal axons seemed to be insensitive to repeated neonatal capsaicin treatment. This study suggests that S100-ir tooth pulp primary neurons are mostly myelinated and that S100-ir unmyelinated axons in the root pulp are preterminal segments of myelinated stem axons. PMID- 9730115 TI - Laminar differences in bicuculline methiodide's effects on cortical neurons in the rat whisker/barrel system. AB - Extracellular unit recordings were made at various depths within SmI barrel cortex of immobilized, sedated rats, in the presence and absence of titrated amounts of the GABA(A) receptor antagonist bicuculline methiodide (BMI). Principal and adjacent whiskers were moved singly, or in paired combination in a condition-test paradigm, to assess excitatory and inhibitory receptive field (RF) characteristics. Neurons were classified as regular- or fast-spike units, and divided into three laminar groups: supragranular, granular (barrel), and infragranular. BMI increased response magnitude and duration, but did not affect response latencies. The excitatory RFs of barrel units, which are the most tightly focused on the principal whisker, were the most greatly defocused by BMI; infragranular units were least affected. All three layers had approximately equal amounts of adjacent whisker-evoked, surround inhibition, but BMI counteracted this inhibition substantially in barrel units and less so in infragranular units. The effects of BMI were most consistent in the barrel; more heterogeneity was found in the non-granular layers. These lamina-dependent effects of BMI are consistent with the idea that between-whisker inhibition is generated mostly within individual layer IV barrels as a result of the rapid engagement of strong, local inhibitory circuitry, and is subsequently embedded in layer IV's output to non-layer IV neurons. The latter's surround inhibition is thus relatively resistant to antagonism by locally applied BMI. The greater heterogeneity of non granular units in terms of RF properties and the effects of BMI is consistent with other findings demonstrating that neighboring neurons in these layers may participate in different local circuits. PMID- 9730116 TI - Impaired detection of repetitive stimulation following interruption of the dorsal spinal column in primates. AB - Transection of the dorsal spinal column in monkeys has been previously shown to spare detection, localization and a variety of discriminations between spatial attributes of tactile stimuli. In contrast, performance on certain tests involving stimulus sequences is substantially impaired, such as tactile direction sensitivity and frequency discrimination. The present study extends these findings to show that a repetitive cutaneous stimulus is undetectable following complete interruption of the ipsilateral dorsal column. Macaca arctoides monkeys were trained to discriminate between different durations of 10 Hz indentation of the glabrous skin of one foot. Preoperatively, these animals could discriminate reliably between three pulses (the standard stimulus duration of 200 ms) and comparison trains of six or more pulses (500 ms or more). Following incomplete interruption of the ipsilateral dorsal column of one monkey, discrimination of the duration of stimulation was unimpaired. However, complete lesions of the ipsilateral dorsal column eliminated performance above the criterion of 75% correct responses for approximately 1 year of postoperative testing of three monkeys. Comparison stimuli of as many as 38 pulses (3.7 s) were utilized during postoperative testing. The inability to detect repetitive stimulation is hypothesized to be related to abnormal intracortical inhibition that has been demonstrated to occur within the primary somatosensory cortex (SI) of monkeys after interruption of the contralateral dorsal column. PMID- 9730117 TI - The use of numbers and percentages in scientific writing. PMID- 9730118 TI - Intraobserver and interobserver reliability of the classification of thoracic adolescent idiopathic scoliosis. AB - The system described by King et al. is the standard method for the classification of thoracic adolescent idiopathic scoliosis. Although it is widely used and referenced, its reliability and reproducibility among scoliosis surgeons are unknown. We used a scoliosis case-presentation format to examine the interobserver and intraobserver reliability of the classification of thoracic adolescent idiopathic scoliosis with the system of King et al. Eight active, current members of the Scoliosis Research Society reviewed twenty-seven full length radiographs that had been made before operative correction of the scoliotic deformity. On the basis of these images, which included posteroanterior and lateral radiographs made with the patient standing as well as right and left forced-side-bending radiographs made with the patient supine, the reviewers assigned a type to each curve according to the classification system of King et al. Kappa coefficients were used to test statistical reliability. The mean interobserver reliability of the classification was only 64 per cent (range, 54 to 77 per cent) when the responses of seven of the reviewers were compared with those of one of the originators of the classification. The mean kappa coefficient was 0.49 (range, 0.27 to 0.73), which indicates poor reliability. When each reviewer's responses were compared with those of the other reviewers, the reliability was similarly poor (interobserver reliability, 55 per cent [range, 33 to 81 per cent] and mean kappa coefficient, 0.40 [range, 0.21 to 0.63]). Intraobserver reliability was evaluated in a trial in which five reviewers in a group setting were shown the same radiographs in a different order at two different viewings. Comparison of the results at the two viewings revealed a mean intraobserver reliability of 69 per cent (range, 56 to 85 per cent) and a mean kappa coefficient of 0.62 (range, 0.34 to 0.95), which indicates fair reliability. The current method of classification of adolescent idiopathic scoliosis does not appear to have sufficient intraobserver or interobserver reliability among scoliosis surgeons to portray curve types accurately. Thus, it may not help to guide treatment with use of modern spinal fixation methods. PMID- 9730119 TI - Interobserver reliability and intraobserver reproducibility of the system of King et al. for the classification of adolescent idiopathic scoliosis. AB - The classification of adolescent idiopathic scoliosis with use of the system of King et al. has become widely accepted since its introduction. The purpose of the present study was to establish the interobserver reliability and intraobserver reproducibility of this classification system. The preoperative radiographs of sixty-three patients who were managed operatively for adolescent idiopathic scoliosis were classified by five observers with the system of King et al. Interobserver reliability was assessed by comparison of the classification of the curves among the observers, and intraobserver reproducibility was evaluated by comparison of the classifications of each set of radiographs by each observer on two occasions three weeks apart. The median interobserver reliability kappa coefficient for the classification system of King et al. was 0.44 (range, 0.28 to 0.50), and the median intraobserver reproducibility kappa coefficient was 0.64 (range, 0.44 to 0.72). According to the definition of Landis and Koch, the classification system of King et al. is substantially reproducible but is only moderately reliable. However, according to the stricter definition of Svanholm et al., its reproducibility is only fair and its reliability is poor. PMID- 9730120 TI - Biomechanical and histological evaluation of a calcium phosphate cement. AB - It is often difficult to achieve stable fixation of a comminuted fracture associated with a metaphyseal defect. The injection of a resorbable cement into an osseous defect may help to stabilize the fracture and to maintain osseous integrity as the cement is resorbed and replaced by bone. The purpose of the present study was to evaluate the repair of a metaphyseal defect after treatment with an injectable calcium-phosphate cement. The injectable cement undergoes isothermic curing in vivo to form a carbonated apatite (dahllite) with a compressive strength of twenty-five megapascals. Either the cement or allograft bone was placed in proximal tibial metaphyseal and distal femoral metaphyseal defects in seventy-two dogs and was evaluated from twenty-four hours to seventy eight weeks postoperatively. Histological examination showed that the cement was osteoconductive; nearly the entire surface area was covered with bone two weeks after the injection. The resulting bone-cement composite underwent gradual remodeling over time in a pattern that was qualitatively similar to the remodeling of normal cortical and cancellous bone. Osteoclasts were found to resorb the cement and were usually associated with adjacent new-bone formation. With increasing time in vivo, the cement was penetrated by small blood vessels that became surrounded by circumferential lamellae of bone and that closely resembled evolving haversian systems. This process occurred more rapidly in the cortex than in the medulla. Mechanical testing showed that, by eight weeks, the tibiae that had been treated with cement had reached nearly 100 per cent of the torsional strength of the contralateral, control (intact) tibiae; this finding paralleled the histological observations of bone apposition to the cement and rapid restoration of the cortex. At no time was fibrous tissue present between the cement and the bone, and there was no evidence of acute inflammation. Small particles of cement were present within occasional macrophages during the process of cement resorption, but the macrophages disappeared over time and were not associated with fibrosis or unexpected resorption of bone. Resorption of the cement was incomplete in the medullary area at seventy-eight weeks, but the pattern of cement resorption and bone-remodeling suggested gradual restoration of a physiological proportion of bone and marrow in both the cortical and the medullary region with maintenance of mechanical function. PMID- 9730121 TI - Teaching medical ethics to orthopaedic surgery residents. AB - Orthopaedic surgery residents will be faced with a variety of ethical issues when they enter clinical practice. A previous survey suggested that they lack knowledge about how to approach several types of medical ethics dilemmas. We developed a medical ethics curriculum for orthopaedic surgery residents and presented it over a one-year period to the residents in one training program. The effect of the educational intervention on the residents' knowledge of medical ethics and their ability to handle hypothetical situations was measured by comparing their responses to a questionnaire, administered before and after the intervention, with those of residents in a training program in which the intervention was not provided. The twenty-five residents at the site of the educational intervention had a mean improvement of 0.10 in the overall score, from a mean score of 0.71 on the baseline survey to a mean score of 0.81 on the follow-up survey. This improvement was significantly greater than the mean improvement of 0.02 for the thirty residents at the control site, who had a mean score of 0.72 on the baseline survey and a mean score of 0.74 on the follow-up survey (p = 0.002). Six residents who participated in the medical ethics curriculum rated it as very useful; seventeen, as somewhat useful; one, as slightly useful; and one, as not at all useful. A medical ethics curriculum can increase orthopaedic residents' knowledge of medical ethics. Whether this curriculum also will lead to behavioral changes requires additional evaluation. PMID- 9730122 TI - Development of a patient-reported measure of function of the knee. AB - The purpose of the present study was to demonstrate the reliability, validity, and responsiveness of the Activities of Daily Living Scale of the Knee Outcome Survey, a patient-reported measure of functional limitations imposed by pathological disorders and impairments of the knee during activities of daily living. The study comprised 397 patients; 213 were male, 156 were female, and the gender was not recorded for the remaining twenty-eight. The mean age of the patients was 33.3 years (range, twelve to seventy-six years). The patients were referred to physical therapy because of a wide variety of disorders of the knee, including ligamentous and meniscal injuries, patellofemoral pain, and osteoarthrosis. The Activities of Daily Living Scale was administered four times during an eight-week period: at the time of the initial evaluation and after one, four, and eight weeks of therapy. Concurrent measures of function included the Lysholm Knee Scale and several global measures of function. The subjects also provided an assessment of the change in function, with responses ranging from greatly worse to greatly better, at one, four, and eight weeks. The Activities of Daily Living Scale was administered to an additional sample of fifty-two patients (thirty-two male and twenty female patients with a mean age of 31.6 years [range, fourteen to sixty-six years]) before and after treatment within a single day to establish test-retest reliability. Factor analysis revealed two dominant factors: one that reflected a combination of symptoms and functional limitations and the other, only symptoms. The internal consistency of the Activities of Daily Living Scale was substantially higher than that of the Lysholm Knee Scale (coefficient alpha, 0.92 to 0.93 compared with 0.60 to 0.73), resulting in a smaller standard error of measurement for the former scale. Validity was demonstrated by moderately strong correlations with concurrent measures of function, including the Lysholm Knee Scale (r = 0.78 to 0.86) and the global assessment of function as measured on a scale ranging from 0 to 100 points (r = 0.66 to 0.75). Analysis of variance with repeated measures revealed significant improvements in the score on the Activities of Daily Living Scale during the eight weeks of physical therapy (F2,236 = 108.13; p < 0.0001); post hoc testing indicated that the change in the score at eight weeks was significantly greater than the change at four weeks and that the change at four weeks was significantly greater than that at one week (p < 0.0001 for both). As had been hypothesized, the patients in whom the knee had somewhat improved had a significantly smaller change in the score, both at four weeks (F1,189 = 33.50; p < 0.001) and at eight weeks (F1,156 = 22.48; p < 0.001), compared with those in whom the knee had greatly improved. The test-retest reliability coefficient (intraclass correlation coefficient[2,1]) was 0.97. These results suggest that the Activities of Daily Living Scale is a reliable, valid, and responsive instrument for the assessment of functional limitations that result from a wide variety of pathological disorders and impairments of the knee. PMID- 9730123 TI - The correlation of comorbidity with function of the shoulder and health status of patients who have glenohumeral degenerative joint disease. AB - We studied the effect of comorbidities on function of the shoulder and health status in a group of eighty-five consecutive patients who had glenohumeral degenerative joint disease of sufficient severity to meet one surgeon's criteria for the performance of shoulder arthroplasty. A questionnaire was used to identify the comorbidities, such as other diseases, social factors, or a work related injury, for each patient. The number of functions on the Simple Shoulder Test that the patient could perform had a significant negative correlation with the number of comorbidities (r = -0.32, intercept = 4.6 per cent, slope = -0.6, and p = 0.0031). Each parameter on the Short Form-36 (except for physical role function) had a significant negative correlation with the number of comorbidities (p < 0.05). This negative relationship was strongest for general health perception (r = -0.42) and vitality (r = -0.35). We concluded that the number of comorbidities has a quantitative effect on function of the shoulder. In the evaluation of the functional status of patients and the effectiveness of treatment, the effects of comorbidity must be controlled. The results of the present study demonstrate that the scores on the Short Form-36 are quantitatively related to the number of comorbidities. The six parameters that are unrelated to function of the shoulder (physical function, social function, emotional role function, mental health, vitality, and general health perception) may provide a practical way to integrate the effects of all potential comorbidities on individual patients. Future clinical research will be strengthened by efforts to measure the impact of comorbidities and by strategies to control for their effects. PMID- 9730124 TI - Kienbock disease and negative ulnar variance. AB - We compared the degree of ulnar variance, measured on standardized radiographs of the wrist, in forty-four patients who had Kienbock disease with that in ninety nine control subjects who had been selected from a general clinic population and had radiographs of the wrist. The purpose of our study was to determine if there is a true relationship between negative ulnar variance and the development of Kienbock disease. Gender was not found to influence the degree of ulnar variance, but an association was found between age and negative ulnar variance in both the control subjects and the patients who had Kienbock disease. The findings of the present study confirmed an association between negative ulnar variance and the development of Kienbock disease even after correction for the influence of age on the measurement of ulnar variance. PMID- 9730125 TI - Comparison of the use of a foot pump with the use of low-molecular-weight heparin for the prevention of deep-vein thrombosis after total hip replacement. A prospective, randomized trial. AB - We conducted a prospective, randomized trial to compare the safety and effectiveness of the A-V Impulse System foot pump with that of low-molecular weight heparin for reducing the prevalence of deep-vein thrombosis after total hip replacement. Of 290 patients who were to have a primary total hip replacement, 143 were randomized to receive enoxaparin (forty milligrams daily) for seven days after the operation and 147, to use the foot pump for seven days. The primary outcome measure was the prevalence of deep-vein thrombosis, as determined by venography on the sixth, seventh, or eighth postoperative day. Secondary outcome measures included transfusion requirements, intraoperative blood loss, postoperative drainage, blood-loss index, appearance of the site of the wound according to a subjective visual-analog scale, and swelling of the thigh. The patients' compliance with the regimen for use of the foot pump was monitored with an internal timing device, and their acceptance of the device was assessed with a questionnaire. Symptoms consistent with pulmonary embolism were investigated with ventilation-perfusion scanning. The patients were contacted later for detection of symptoms of venous thromboembolism that may have occurred during the first three months after discharge from the hospital. Venography was performed on 274 patients: 136 who used the foot pump and 138 who received enoxaparin. Deep-vein thrombosis was detected in twenty-four (18 per cent) of the patients who used the foot pump compared with eighteen patients (13 per cent) who received enoxaparin (95 per cent confidence interval for the difference in proportions, -3.9 to +13.0 per cent). Thrombosis in the calf was found in seven patients (5 per cent) in the former group compared with six patients (4 per cent) in the latter (95 per cent confidence interval for the difference, -4.2 to +5.8 per cent), and proximal thrombosis was observed in seventeen patients (13 per cent) in the former group compared with twelve patients (9 per cent) in the latter (95 per cent confidence interval for the difference, -3.5 to +11.1 per cent). None of these differences was significant. No patient in either group had major proximal deep-vein thrombosis; all proximal thrombi were isolated entities involving the femoral valve cusp and were of unknown importance. One patient who used the foot pump had a non-fatal pulmonary embolism. One patient who received enoxaparin had a symptomatic deep-vein thrombosis during hospitalization. Two patients (one from each group [0.7 per cent]) were readmitted to the hospital because of a symptomatic deep-vein thrombosis despite normal venographic findings at the time of discharge. There was no difference in the transfusion requirements or the intraoperative blood loss between the two groups. There were more soft tissue side effects in the patients who received enoxaparin than in those who used the foot pump: there was more bruising of the thigh and oozing of the wound (p < 0.001 for each), postoperative drainage (578 compared with 492 milliliters; p = 0.014), and swelling of the thigh (twenty compared with ten millimeters; p = 0.03). Of 124 patients who used the foot pump and were asked about the acceptability of the device, fourteen (11 per cent) said that it was uncomfortable, twenty-one (17 per cent) reported sleep disturbance, and four (3 per cent) stated that they had stopped using the device. Conversely, ten (8 per cent) found it relaxing. We concluded that the foot pump is a suitable alternative to low-molecular-weight heparin for prophylaxis against thromboembolism after total hip replacement and that it produces fewer soft tissue side effects. Tolerance of the device is a problem for some patients. PMID- 9730126 TI - Ultrasound surveillance for asymptomatic deep venous thrombosis after total joint replacement. AB - Prospective data on 202 consecutive patients who had a total of 123 total hip and ninety-four total knee arthroplasties were collected from two university medical centers. The findings of routine surveillance for deep venous thrombosis performed with ascending contrast venography were compared with those of surveillance with duplex ultrasonography complemented with color-flow Doppler imaging. All of the studies were performed between the third and seventh postoperative days. Of the 202 patients (342 extremities) who were examined, fifty-five (27 per cent) were found to have deep venous thrombosis; fifty-two (95 per cent) of the thrombi were in the calf and three (5 per cent) were in the proximal veins. All of the thrombi were clinically asymptomatic and all were nonocclusive, allowing passage of contrast medium around an intraluminal filling defect. Duplex ultrasonography with color-flow Doppler imaging correctly identified two of the three proximal thrombi and five of the fifty-two thrombi in the calf (sensitivity, 10 per cent). The sensitivity for the detection of thrombi in the calf was zero of sixteen at one of the institutions involved in the study and 14 per cent (five of thirty-six) at the other. There were two false-positive findings on ultrasonographic examination; one involved a proximal thrombus and one, a distal thrombus. We believe that the interinstitutional variability and insensitivity of duplex ultrasonography with color-flow Doppler imaging for the detection of asymptomatic deep venous thrombi in the calf after total joint replacement make it unreliable as a routine surveillance tool after total hip or knee arthroplasty. PMID- 9730127 TI - Fixation of acetabular cups without cement in total hip arthroplasty. A comparison of three different implant surfaces at a minimum duration of follow-up of five years. AB - We evaluated 377 patients (428 hips) who had been managed, by a total of fourteen surgeons at twelve clinical sites in the United States and Europe, with a porous coated press-fit acetabular cup, a hydroxyapatite-coated threaded screw-in cup, or one of two similar designs of hydroxyapatite-coated press-fit cups between April 1987 and November 1992. The same type of hydroxyapatite-coated femoral stem was inserted without cement in all patients. After a minimum duration of follow up of five years (mean, 7.9 years; range, 5.3 to 9.1 years), one (1 per cent) of the 131 hydroxyapatite-coated threaded cups, two (2 per cent) of the 109 porous coated press-fit cups, and twenty-one (11 per cent) of the 188 hydroxyapatite coated press-fit cups had been revised because of aseptic loosening. A common radiographic sign of impending failure of the hydroxyapatite-coated press-fit cups was radiolucency at the interface between the implant and the subchondral bone beneath it. This radiolucency usually was seen initially more than two years after implantation. Radiographic evaluation of the 383 acetabular implants that were in situ at the time of the most recent follow-up showed that 123 (99 per cent) of the 124 hydroxyapatite-coated threaded cups, 101 (98 per cent) of the 103 porous-coated cups, and 139 (89 per cent) of the 156 hydroxyapatite-coated press-fit cups were stable with osseous ingrowth (as indicated by the absence of radiolucency at the interface and the absence of migration within the acetabulum). The probability of revision due to aseptic loosening was significantly greater for the hydroxyapatite-coated press-fit cups than it was for the hydroxyapatite-coated threaded cups or the porous-coated press-fit cups (p < 0.001 for both comparisons). Within the group of patients who had a hydroxyapatite-coated press-fit cup, the probability of revision due to aseptic loosening was significantly greater in association with a young age (p = 0.003), female gender (p = 0.02), the use of a femoral head with a diameter of thirty-two millimeters (p = 0.018), and the use of a thin polyethylene liner (p < 0.001). We found that the hydroxyapatite-coated threaded cups and the porous-coated press fit cups continued to perform well more than five years after the operation. The hydroxyapatite-coated press-fit cups that were revised probably failed because the fixation interface beneath the cup could not sustain the tensile stresses that were imposed between the cup and the bone by the activity of the patient. Our data suggest that, in the specific biomechanical environment of the acetabulum, physical interlocking between the cup and the supporting bone beneath it may be a prerequisite for long-term stability. PMID- 9730128 TI - Mobilization of a congenital proximal radioulnar synostosis with use of a free vascularized fascio-fat graft. AB - We present the results of a new mobilization procedure for the treatment of a congenital proximal radioulnar synostosis in seven patients. The operative procedure included separation of the synostosis and placement of a free vascularized fascio-fat graft to prevent recurrent ankylosis. The average age at the time of the operation was eight years and two months (range, six years and four months to eleven years and ten months). All of the patients were boys who had no other congenital anomalies. The radial head was dislocated in all seven patients (anteriorly in two and posteriorly in five). The final four index operations included an osteotomy of the radius in order to reduce the dislocated radial head. The average duration of follow-up was three years and eight months (range, two years and four months to four years and five months). Preoperatively, the patients had had difficulty with holding a bowl of soup and accepting objects, such as coins, into the palm. Postoperatively, they were able to perform these activities. None of the patients had recurrent ankylosis or loss of the flap. The average supination was 26 degrees (range, 10 to 45 degrees), and the average pronation was 45 degrees (range, 10 to 80 degrees). The four patients who had had an osteotomy of the radius in addition to the index procedure did not have a dislocation of the radial head and had an average arc of motion of 83 degrees of pronation and supination. The three patients who had not had an osteotomy had a dislocation of the radial head and an average arc of motion of 40 degrees after the index procedure. These findings demonstrate that separation of a congenital radioulnar synostosis with a vascularized fascio-fat graft and osteotomy of the radius can achieve pronation and supination of the forearm. PMID- 9730129 TI - Calcific myonecrosis mimicking an invasive soft-tissue neoplasm. A case report and review of the literature. PMID- 9730130 TI - Probable interspousal transmission of Staphylococcus aureus resulting in joint infection after total knee arthroplasty. A report of two cases. PMID- 9730131 TI - Spontaneous healing of a tear of the anterior cruciate ligament. A report of two cases. PMID- 9730132 TI - Evaluation and staging of musculoskeletal neoplasms. PMID- 9730133 TI - The multifactorial nature of polyethylene wear in vivo. PMID- 9730134 TI - School-screening for scoliosis. A prospective epidemiological study in northwestern and central Greece. PMID- 9730135 TI - Acute tubular necrosis of an allograft kidney following total hip replacement. A case report. PMID- 9730136 TI - A comparison of fixation screws for the scaphoid during application of cyclical bending loads. PMID- 9730137 TI - Prevention of deep-vein thrombosis after total hip arthroplasty. Comparison of warfarin and dalteparin. PMID- 9730138 TI - Prevention of deep-vein thrombosis after total hip arthroplasty. Comparison of warfarin and dalteparin. PMID- 9730139 TI - Genetic polymorphisms and mutations of the lipoprotein lipase gene in Japanese schoolchildren with hypoalphalipoproteinemia. AB - Lipoprotein lipase (LPL) is an important enzyme for the hydrolysis of TG on lipoproteins, and its activity is positively correlated with the plasma levels of high density lipoprotein cholesterol (HDL-C). To investigate the association between the LPL gene and low HDL-C levels, we studied two polymorphisms (Hind III and Pvu II) and three mutations (Asn291Ser, Gly188Glu and LPL(Arita)) of the LPL gene in 114 children with low HDL-C levels (<40 mg/dl) and 194 control children using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) techniques (PCR-RFLP). The frequency of the Pvu II +/+ genotype was significantly higher in the children with low-HDL/high-TG (TG>100 mg/dl, 90th percentile level among Japanese schoolchildren) than in the other children (vs the low-HDL/normal-TG children, chi2 = 7.49, p < 0.01; vs control children, chi2 = 7.23, p < 0.01). Pvu II+ allele of the LPL gene was associated with elevated TG levels in low HDL-C groups. In addition, we found one heterozygote of LPL(Arita) (deletion of G at base 916 in exon 5, the most common mutation of LPL deficiency in Japanese), among the low-HDL/high-TG subjects. The other two variants were not detected in either the low-HDL children or control children. LPL Asn291Ser and Gly188Glu have been presumed to be rare in the Japanese population. In conclusion, our results suggest that hypoalphalipoproteinemia with elevated TG level may be associated with genetic variations of the LPL gene. PMID- 9730140 TI - Simvastatin, a potent HMG-CoA reductase inhibitor, inhibits the proliferation of human and bovine endothelial cells in vitro. AB - We investigated in vitro effect of simvastatin, a potent 3-hydroxy-3 methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor, on the proliferation of human and bovine endothelial cells (EC) compared to that of bovine smooth muscle cells (SMC). Cells were cultured in a medium supplemented with 10% normal serum (FBS). Simvastatin at concentrations ranging from 0.1 microg/ml to 10 microg/ml were used. In each kind of cells the proliferation was markedly reduced at 1 microg/ml of simvastatin (P<0.01) which was accompanied by morphological changes in the cell shape. We conclude that simvastatin inhibits the proliferation of not only the smooth muscle cells but also the endothelial cells. PMID- 9730141 TI - Multi-modal expression of apolipoprotein (a) gene in vivo. AB - The apolipoprotein (a) [apo(a)] gene encodes a protein component of lipoprotein (a) [Lp(a)] whose plasma levels vary among individuals. To study the implications of Lp(a), we examined plasma Lp(a) levels and molecular weights of apo(a) in patients with cerebrovascular disease (CVD) or diabetes mellitus (DN). Mean Lp(a) concentrations were higher in the CVD cases with atherothrombotic brain infarction than in those with brain hemorrhage and lacunar infarction. Lp(a) levels were lower in the DM cases on diet therapy alone than in those treated with insulin or oral hypoglycemic agents. These results suggest that Lp(a) is thrombogenic and atherogenic, and that insulin may modulate Lp(a) levels. We subclassified the apo(a) gene into four types (A-D) by polymorphisms in the 5' flanking region. We also measured plasma Lp(a) concentrations and examined expression of the gene by an in vitro assay. Homozygotes of type C had higher Lp(a) levels than those of type D, and the relative expression of type C was higher than that of type D in vitro. Lp(a) levels, however, varied even within the same 5'-allele having similar apo(a) isoforms. Thus, Lp(a) concentrations are genetically determined and may be modified by some hormones and cytokines. When we examined transcript levels for apo(a) by RT-PCR in various normal tissues, apo(a) was strongly expressed in liver while not in thyroid or leukocytes. Small amounts of apo(a) transcript were observed in all other organs and tissues. Apo(a) in these tissues may also play a role in inframmation, tissue remodeling, cell migration, and other physiological functions. PMID- 9730143 TI - Endothelial dysfunction in hypertension. AB - The endothelium modulates the tone of the underlying vascular smooth muscle by releasing relaxing factors, including prostacyclin, nitric oxide (NO), and endothelium-derived hyperpolarizing factor (EDHF). In most types of hypertension, endothelium-dependent relaxations are impaired because of a reduced production and/or action of endothelium-derived NO and EDHF. In essential hypertension, endothelium-dependent relaxations are reduced because of a concomitant release of vasoconstrictor prostanoids (endoperoxides and thromboxane A2). These prostanoids may be produced in the vascular smooth muscle rather than in the endothelium. The endothelial dysfunction observed in hypertension is likely to be a consequence rather than a cause of the disease, representing premature aging of the blood vessels due to the chronic exposure to the high blood pressure. The endothelial dysfunction can be improved by antihypertensive therapy, favoring the prevention of the occurrence of vascular complications in hypertension. PMID- 9730142 TI - A candidate high density lipoprotein (HDL) receptor, HB2, with possible multiple functions shows sequence homology with adhesion molecules. AB - High density lipoprotein (HDL), an antiatherogenic lipoprotein comprises several subclasses differing in composition and metabolic function. This physiological complexity appears to be matched with the growing number of candidate HDL receptors, since several HDL binding proteins have recently been identified in various tissues. Because these putative receptors may signal different pathways it is important to identify their structure and potential role in HDL metabolism. We review recent progress in the cloning and characterization of HB2, one of a pair of HDL binding proteins (HB1 and HB2) first purified from rat liver. The structure of HB2 is consistent with a receptor role for this membrane protein and when expressed in cells, increases HDL binding 2 fold. HB2, minimally present in THP-1 cells, is substantially upregulated in macrophages and appears sensitive to cholesterol loading of these cells. The protein shows high homology with adhesion molecules ALCAM and BEN and the possibility that HDL by binding to HB2, reduces adhesion induced arterial wall injury, is discussed in the context of the known protective role of HDL against atherosclerosis. The possible functions of HB2 are also compared with other recently cloned HDL receptors. PMID- 9730144 TI - Contribution of a vascular modulator, hepatocyte growth factor (HGF), to the pathogenesis of cardiovascular disease. AB - HGF is a mesenchyme-derived pleiotropic factor which regulates cell growth, cell motility, and morphogenesis of various types of cells, and is thus considered a humoral mediator of epithelial-mesenchymal interactions responsible for morphogenic tissue interactions during embryonic development and organogenesis. Although HGF is originally identified as a most potent mitogen for hepatocytes, HGF is also belonged to a member of endothelium-specific growth factors. Since endothelial cells are known to secrete various anti-proliferative and vasodilating factors, an agent that promotes seeding or regeneration of endothelium may have potential therapeutic value against vascular smooth muscle cell proliferation. The mitogenic action of HGF on human endothelial cells was most potent among growth factors. Moreover, the presence of local HGF system (HGF and its specific receptor, c-met) was observed in vascular cells and cardiac myocytes in vitro as well as in vivo. Production of local HGF production in vascular cells was regulated by various cytokines including transforming growth factor (TGF)-beta and Ang II. Furthermore, HGF may be therapeutic growth factors for the treatment of restenosis after angioplasty and arteriosclerosis oblerance, etc., as gene therapy. On the other hand, serum HGF concentration was significantly correlated with blood pressure. These results suggest that HGF secretion might be elevated in response to high blood pressure as a counter system against endothelial dysfunction, and may be considered as an index of severity of hypertension. In this review, we discussed the potential role of HGF in cardiovascular disease. PMID- 9730146 TI - Case of the month: small rectal carcinoma with lymph node and liver metastases. PMID- 9730145 TI - The role of tissue factor in the pathogenesis of thrombosis and atherosclerosis. AB - TF is a major regulator of coagulation and hemostasis. High levels of TF antigen and activity are detected in atherosclerotic lesions, particularly in the advanced lesions. When the plaques are ruptured or eroded, exposure of cellular and extracellular TF to circulating blood play a pivotal role in mediating fibrin rich thrombus formation leading to acute coronary syndromes. On the other hand, activation of blood coagulation and deficiency of coagulation inhibitors, without endothelial cell denudation, are considered to be an important factor of thrombogenesis in the microcirculation. The imbalance between TF and TFPI seems to be important in promoting fibrin thrombus formation in the lung of endotoxin induced DIC condition. PMID- 9730147 TI - Treatment of metastatic breast cancer patients: what is the standard care of the patients? PMID- 9730148 TI - Transitional cell carcinoma of the urethra in men and women associated with bladder cancer. AB - Multifocal tumor occurrence in the entire urinary tract in time and space is a well-recognized characteristic of transitional cell carcinoma. Synchronous and asynchronous urethral transitional cell carcinoma, in relation to bladder cancer in male and female patients, is the subject of the present mini-review. It is imperative to rule out male and female patients having a high risk for urethral involvement or urethral recurrence. In male patients, prostatic urethral involvement and stromal invasion mainly due to in situ extension of carcinoma seems to be the most important risk factor. In female patients, bladder neck involvement by cancer seems most important. By excluding male and female bladder cancer patients having these characteristics for simultaneous urethrectomy, other patients are good candidates for reconstruction of the urinary tract after cystectomy by an orthotopic neobladder which will offer a good quality of life to bladder cancer patients. PMID- 9730149 TI - Effects of soybean isoflavones on cell growth and apoptosis of the human prostatic cancer cell line LNCaP. AB - BACKGROUND: Epidemiological studies have suggested that soybean isoflavones are associated with a lower risk of prostate cancer. However, the mechanisms of prostate cancer prevention by soybean isoflavones have yet to be fully clarified. METHODS: Two soybean isoflavones (genistein and daidzein) and their glucosides (genistin and daidzin) were tested for their effects on cell growth and apoptosis of the LNCaP human prostatic cancer cell line. RESULTS: Among these isoflavones, genistein was found to inhibit the growth of LNCaP most effectively, with an IC50 value of 40 microM. The inhibition of cell growth by genistein was accompanied by the suppression of DNA synthesis and the induction of apoptosis. Expression of prostate-specific antigen (PSA) in LNCaP was also significantly reduced by the treatment with genistein. CONCLUSIONS: The results suggest that genistein might primarily influence human prostate cancer development by reducing tumor growth. PMID- 9730150 TI - Radiotherapy in the management of Graves' ophthalmopathy. AB - BACKGROUND: To report the results of radiotherapy for patients with failure, adverse reactions or relative contraindications to the use of steroids or immunosuppressants, by using newly developed quantitative indexes. METHODS: Fourteen female and six male patients with Graves' ophthalmopathy were treated with radiotherapy between 1989 and 1996. Prior to radiotherapy, eight patients received treatment with prednisone, four received immunosuppressants and four received a combination of both. Four patients with contraindications to steroids were initially managed with radiotherapy. Most of the patients received a dose of 24-28 Gy in 2 Gy fractions. We used the newly developed motility limitation index to assess extraocular motility. RESULTS: Treatment was well tolerated. There have been no late complications. All 12 patients with soft tissue signs such as edema, irritation, tearing and pain were improved. Proptosis did not improve or improved only slightly, 3 mm at best. However, proptosis in all but two has been stabilized and has not deteriorated in the follow-up period. Most of the patients have experienced an improvement of eye-muscle motility. Extraocular muscles that work for elevation were impaired more severely than the other muscles and this tended to remain. Of the 16 patients using steroids before or when radiotherapy was initiated, 15 were tapered off and only one patient required additional steroids, thus sparing the majority from steroid adverse reactions. CONCLUSION: Radiotherapy was effective in preventing exacerbations of active inflammatory ophthalmopathy in patients with Graves' disease with minimal morbidity and thus eliminated the adverse reactions associated with protracted corticosteroid use. The newly developed motility limitation index was useful in detecting delicate changes in motility of individual extraocular muscles. PMID- 9730151 TI - Clinical characteristics of patients with metastatic breast cancer with complete remission following systemic treatment. AB - BACKGROUND: Patients with metastatic breast cancer (MBC) have variable clinical courses. The purpose was to describe the clinical characteristics of MBC patients with complete remissions (CR) following systemic treatment. METHODS: We analyzed 315 consecutive MBC patients treated with several types of systemic treatments at the National Cancer Center Hospital between January 1988 and December 1993. RESULTS: The median survival time (MST) and median progression-free survival were 28.0 and 17.1 months, respectively. Forty patients were defined as 'first-CR' following initial or second-line systemic treatment and the majority of them had a good performance status, low number of metastatic sites and low incidence of liver involvement. Nine of 40 patients with first-CR continued progression-free 5 years after beginning systemic treatments. The major sites of metastasis were the lung and bone and there were no cases with liver metastasis. Five patients received standard doxorubicin-containing combination chemotherapy with or without tamoxifen. Two of these nine patients remain progression free in first-CR. Three of them remained in first-CR after 5 years and died of progressive breast cancer and two others died of unrelated causes. Two patients relapsed after obtaining a first-CR for at least 5 years and remain alive with active metastatic disease. The MST and median progression-free survival of nine patients were 10.6 and 9.0 years, respectively. These nine patients represented 22.5% of all first-CR patients and 3.2% of the total patients. CONCLUSIONS: Although MBC is commonly recognized to be an incurable disease, a small percentage of patients clearly are alive and progression free for prolonged periods after initiation of systemic treatments. PMID- 9730152 TI - Low-dose cisplatin-5-fluorouracil prevents postoperative suppression of natural killer cell activity in patients with gastrointestinal cancer. AB - BACKGROUND: The effect of administering low-dose cisplatin (CDDP)-5-fluorouracil (5-FU) postoperatively on the activity of the immune system is not known. To clarify the effect on natural killer (NK) cell activity of treatment with low dose CDDP-5-FU, we compared NK cell activity after surgery for gastrointestinal cancer in patients treated with low-dose CDDP-5-FU, a bolus dose of mitomycin C (MMC) or no anticancer drug. METHODS: Sixty-two patients consisted of three groups: low-dose CDDP-5-FU (n = 15), MMC (n = 20) and no-drug (n = 27). Chemotherapy was initiated immediately after surgery. NK cell activity was measured on the day before surgery (pre-op) and on postoperative days 7 (POD7) and 21 (POD21). RESULTS: The NK cell activities of the CDDP-5-FU group were 37.7% at pre-op, 36.1% on POD7 and 33.6% on POD21. However, the NK cell activities in the no-drug and MMC groups were significantly decreased on POD7 (from 36.6 to 24.8% and from 31.4 to 16.6%, respectively). The NK cell activity in the MMC group remained depressed on POD21 (18.6%) whereas that in the no-drug group recovered (31.6%). CONCLUSIONS: Consecutive administration of low-dose CDDP-5-FU appears to be useful as postoperative adjuvant chemotherapy because of its preventive effect on NK cell suppression after surgery. PMID- 9730153 TI - Clinicopathological characteristics of skipping lymph node metastases in patients with colorectal cancer. AB - BACKGROUND: We have sometimes experienced cases of colorectal cancer with skipping lymph node metastasis in which distant nodes were positive but those closer to the tumor were negative. There have been few reports of this condition and its clinical characteristics have not been clarified. This study was conducted to clarify the status of skipping lymph node metastasis and its clinicopathological characteristics in colorectal cancer. METHODS: We analyzed 452 patients with colorectal cancer and nodal metastases (270 with colon cancer and 182 with rectal cancer). All the resected nodes were examined using histological procedures with a microscope and were classified by their location according to the General Rules for Clinical and Pathological Studies on Cancer of the Colon, Rectum and Anus. We studied the status of skipping nodal status and the correlation between the nodal status and clinicopathological findings, including the disease-free survival, depth of tumor, histological type, staging and recurrence. RESULTS: Twenty-eight (10.4%) of the colon cancer patients and 20 (11.0%) of the rectal cancer patients were found to have skipping nodal metastases. In rectal cancer patients with n2 (nodal metastases at the N2 site) in the direction of the main node, patients with skipping lymph node metastases had a significantly better prognosis than those without (p = 0.026). In all colon cancer patients and rectal cancer patients with lateral n3 (nodal metastases at the lateral N3 site), there were a tendency for those with skipping nodal metastases to have better disease-free survival rates (p = 0.1). Also, the mean number of positive nodes in skipping cases was significantly lower than that in non-skipping cases. In addition, skipping nodal metastases in rectal cancer suggested a possibility of bypass flow which was not generally recognized. CONCLUSION: These findings in colorectal cancer suggest the presence of previously unknown lymphatic tracts and that the cancers concerned have a better prognosis than those without skipping nodal metastases. PMID- 9730154 TI - Point mutations of ornithine decarboxylase gene are an infrequent event in colorectal cancer but a missense mutation was found in a replication error positive patient with hMSH2 germline mutation. AB - BACKGROUND: Ornithine decarboxylase (ODC; EC 4.1.1.17) is the first rate-limiting enzyme in the biosynthesis of polyamines. ODC protein has a characteristic amino acid sequence, the PEST sequence, which is related to the enzyme's rapid degradation. ODC cDNA prepared from human hepatoma tissues has been reported to show nonsense or missense mutations. METHODS: We examined somatic mutations of ODC cDNA by RT-PCR-SSCP analysis and mRNA expressions by RT-PCR in 50 colorectal cancer tissues to investigate the involvement of ODC gene alterations in colorectal cancers. RESULTS: Increased expression of the ODC gene was observed in 36 cases (86%) out of the 42 examined by RT-PCR. In one case, a missense mutation was found in the cancer tissue but not in normal mucosa. The missense mutation from Asp to Asn at codon 424, in the PEST region, possibly stabilizes the ODC protein. In colorectal cancer, replication error and a germline mutation in hMSH2 gene were observed. CONCLUSIONS: The missense mutation at codon 424 is speculated to be a cause of stabilization and a passenger mutation owing to the mutator phenotype. Since only one of 50 colorectal cancers exhibited a missense mutation of the ODC gene, mutations in ODC gene are not frequent in colorectal cancer. The increased expression of the ODC gene was noted in 86% of colorectal cancer tissues by RT-PCR, however, it was not due to point mutations in ODC coding exons. PMID- 9730155 TI - Patterns of care study of radiation therapy for cervix cancer in Japan: the influence of the stratification of institution on the process. AB - BACKGROUND: To improve the quality of radiation oncology in Japan, Patterns of Care Study (PCS), a widely known quality assurance (QA) program in the USA, was introduced. The feasibility was tested by collecting nationwide data by extramural audit for cervix cancer. METHODS: From July 1996 through February 1997, PCS audits were performed for 29 institutions nationwide. On the basis of the facility survey by Tsunemoto, 13 institutions were classified as A1 (university hospital/cancer center), 10 as B1 (other institutions treating >120 patients/year) and six as B2 (other institutions treating <120 patients/year). Medical charts for the patients treated for cervix cancer between 1992 and 1994 were reviewed based on the data format of the US PCS. The total number of patients surveyed was 432. RESULTS: Simulation was used for >90% of the patients in both A1 and B1-2 institutions. However, in B1-2, planning for 5% of the patients was performed with only a clinical set-up (p = 0.0287). A daily fraction with a size of 200 cGy was given to >65% of patients in A1 and to <47% in B1-2. On the other hand, >50% of those in B1-2 were treated with daily fractions of 180 cGy and less compared with 25% in A1 institutions (p < 0.0001). Brachytherapy was utilized more frequently for patients in Stages II (p = 0.0365), III (p = 0.0015) and IV (p = 0.0483) in A1 than in B1-2. As for external beam equipment, linear accelerators with 10 MV or more were used for 83% of the patients in A1. However, in B1-2 institutions, machines with lower energy were used for 38% of the patients (p < 0.0001). The median number of full-time-equivalent (FTE) radiation oncologists was 2.7 in A1, 0.65 in B1 and 0.2 in B2. CONCLUSIONS: Institutional stratification, including equipment and personnel, was found to affect significantly the patterns of care for cervix cancer. Therefore, to improve the quality of radiation therapy nationwide, improvements in equipment and in supply of FTE personnel are extremely important. PCS was found to have great potential for a practical evaluation of how much improvement will be required in Japan. PMID- 9730156 TI - Dual rearrangement of immunoglobulin and T-cell receptor genes in a case of Philadelphia chromosome-positive acute leukemia. AB - A 48-year-old patient was admitted to our hospital for leukocytosis. The blast cells were positive for peroxidase and he was tentatively diagnosed as acute myeloid leukemia according to the French-American-British criteria. By flow cytometry, the bone marrow cells were positive for CD10, CD13, CD33 and HLA-DR, but two-color analysis revealed that most of the CD13- and CD33-positive cells did not express CD10. The marrow cells had Philadelphia chromosome with no additional abnormalities. Major bcr-abl fusion gene was observed by the reverse transcriptase-polymerase chain reaction method. Southern blot analysis disclosed rearrangement of both immunoglobulin heavy chain and T-cell receptor beta chain genes. He received combined chemotherapy for myeloid lineage and lymphoid lineage, but the response was quite poor. He died 64 days after admission due to pulmonary bleeding. Although the association of Ph1 with multilineage differentiation is unclear, our case has significant implication for further investigation of the relationship between Ph1-positive cells and lineage selection. PMID- 9730159 TI - The "Sufficiency Principle" from the perspective of cancer prevention. PMID- 9730158 TI - Primary unknown cancer in pulmonary hilar lymph node with spontaneous transient regression: report of a case. AB - A 69-year-old man was referred to our hospital in December 1993 because an abnormal mass had been detected in the right pulmonary hilum. Computed tomography (CT) of the chest revealed a swollen hilar lymph node between the right middle and lower lobe bronchi, and an adherent tumor in the right ventrobasal segment (S8). Chest roentgenogram in February 1994, however, showed no evident tumor in the right lung field. In March 1996, the mass in the right pulmonary hilum reappeared on chest roentgenogram. Chest CT revealed a swollen hilar lymph node between the right middle and lower lobe bronchi, but there was no tumor in right S8. The patient underwent video-assisted thoracoscopy on 17 May 1996. Intraoperative needle biopsy of the node revealed cancer cells. We performed right middle and lower bilobectomy with mediastinal dissection. Histological diagnosis revealed a large cell carcinoma almost completely occupying a hilar lymph node. The resected middle and lower lobes showed no tumors, except for a coagulation necrosis measuring 1.5 cm in diameter in S8b, corresponding to the site where a tumor shadow had been depicted on the CT image in December 1993. We concluded that the coagulation necrosis might have been the primary site of the tumor, which had spontaneously regressed and then appeared in the metastatic interlobar node. PMID- 9730157 TI - Undifferentiated carcinoma of the liver with neuroendocrine features: a case report. AB - Undifferentiated carcinoma of the liver is very rare. A 54-year-old man was admitted to our hospital for a detailed examination of multiple liver tumors. These tumors were high or low echoic on ultrasonography, but not enhanced by contrast medium in dynamic computed tomography. A fine-needle aspiration biopsy specimen of the tumor showed undifferentiated carcinoma. The serum level of neuron-specific enolase was high (357 ng/ml) and the immunohistochemical stain of the biopsy specimen was positive for synaptophysin. We diagnosed the patient as having undifferentiated carcinoma of the liver with neuroendocrine features. The patient was treated with combined systemic chemotherapy: etoposide 100 mg/m2/day for three days plus cisplatin 80 mg/m2/day on day one. He achieved a partial response, the duration of which was 7+ months. The serum neuron-specific enolase levels were decreased to the normal range after chemotherapy. Primary liver carcinoma with neuroendocrine features is extremely rare, but in a suspicious case it is important to measure the serum levels of neuroendocrine markers and make a histological confirmation, because chemotherapy may be effective for this disease. PMID- 9730161 TI - UK-Japan Workshop on Gastrointestinal Cancer was held towards improved understanding between east and west. PMID- 9730160 TI - The pre-conceived British beliefs of gastric cancer in Japan need to be changed. PMID- 9730162 TI - Breast Cancer Prevention Trial shows major benefit, some risk. PMID- 9730163 TI - Starch granules: structure and biosynthesis. AB - The emphasis of this review is on starch structure and its biosynthesis. Improvements in understanding have been brought about during the last decade through the development of new physicochemical and biological techniques, leading to real scientific progress. All this literature needs to be kept inside the general literature about biopolymers, despite some confusing results or discrepancies arising from the biological variability of starch. However, a coherent picture of starch over all the different structural levels can be presented, in order to obtain some generalizations about its structure. In this review we will focus first on our present understanding of the structures of amylose and amylopectin and their organization within the granule, and we will then give insights on the biosynthetic mechanisms explaining the biogenesis of starch in plants. PMID- 9730164 TI - Formation of a ternary complex between chitosan and ion pairs of strontium carbonate. AB - This work deals essentially with the study of the interactions between chitosan and an alkaline-earth metal: 85Sr. We showed that addition of carbonate ions is necessary to observe this interaction, but only for pH values over 11. The use of an appropriate calculation software allowed us to determine the nature of the strontium ionic species present in solution. In order to understand the mechanism of interaction, the influence of various parameters on the complexation was studied. Thus, the interaction was maximum for a CO3(2-) concentration close to 10(-2) mol/l, the lowest ionic strength, the most expanded physical form (lyophilisate) and the lowest degree of acetylation of the polymer. We conclude to the formation of a ternary complex between the ion pair Sr2+ CO3(2-) and the amino groups of chitosan. We also showed the presence of interactions between chitosan and carbonate ions which hinder further interactions with strontium ions. PMID- 9730165 TI - Interactions between chitosan and alpha emitters: 238Pu and 241Am. AB - This paper deals with the first study published on the interactions between chitosan and two radioactive elements (alpha emitters): 238Pu and 241Am. The authors determined the optimal experimental conditions for each of them (pH, ionic strength (FI), carbonate concentration) and attempted to explain the mechanism of interaction. The study of the nature and the percentage of the metallic ionic species present in solution as a function of the experimental conditions was carried out by means of the TOT software for 241Am. The authors showed a good fixation of 238Pu and 241Am on chitosan for pH's between 11-12, with the best results for 241Am. Nevertheless, an increase of the FI or the addition of a complexant of these alpha emitters induced a decrease of the interactions with chitosan. PMID- 9730166 TI - Alkaline-induced unfolding and salt-induced folding of pig heart lactate dehydrogenase under high pH conditions. AB - The alkaline-induced unfolding and the salt-induced folding of pig heart lactate dehydrogenase under high pH conditions have been followed by fluorescence emission spectra and circular dichroism spectra. The results for alkaline-induced denaturation of lactate dehydrogenase show that at low ionic strength, increasing the pH value increased the extent of unfolding of the enzyme to the maximum ultimate unfolded conformation at about pH 13.0. At pH 12.5, although the enzyme was completely inactivated, most of the ordered structure was retained. Even at pH 13.5, the apparently fully unfolded enzyme still retained some ordered secondary structure. Kinetic analysis showed that at high pH, the inactivation rate constants of the enzyme are an order of magnitude faster than the unfolding rate constants at least. The above results are in accord with the suggestion by Tsou (Trends Biochem Sci 1986;11:427-429 and Science 1993;262:380-381) that the active site is usually more flexible than the enzyme molecule. At pH 13.0, adding salt to the solution caused the relatively unfolded state of the denatured enzyme to change into a compact conformational state by hydrophobic collapsing. The folded state induced by the salt bound ANS strongly, indicating the existence of an increased hydrophobic surface. The above results suggest that the salt-induced folded state at high pH may be the folded intermediate which exists in the general protein folding and that the large residual ordered secondary structure might become folded during the salt-induced folding. PMID- 9730168 TI - Some aspects of beta-lactoglobulin structural properties in solution studied by fluorescence quenching. AB - The technique of protein fluorescence quenching by acrylamide and sodium nitrite (NO2-) was used to study some structural aspects of beta-lactoglobulin in solution. The degree of exposure and the micro-environments of the two tryptophanyl residues (Trp-19 and Trp-61) present in this ruminant milk protein were sensed, and the influence of the pH and the binding of palmitic acid in their accessibilities were analyzed. The results obtained showed that Trp-19 has an accessibility to the quenchers higher than could be supposed from its structural location. The binding of palmitic acid, on the other hand, increases the accessibility of both tryptophanyl residues, a fact that could be associated with a slight conformational change of the protein. PMID- 9730167 TI - Effect of temperature on the intrinsic viscosity and conformation of chitosans in dilute HCl solution. AB - The effects of temperature on the intrinsic viscosity and on the conformation of chitosans in dilute HCI solution were studied. Ten chitosans with the same degree of deacetylation but different molecular weights were produced by alkali deacetylation of chitin which was prepared from red shrimp wastes. The degree of deacetylation at 83% and weight average molecular weight of the chitosans ranging 78-914 kDa were determined by infrared spectroscopy and static light scattering, respectively. The intrinsic viscosities ([eta]) of these 10 chitosans in 0.01 M hydrochloric acid were measured at 10, 20, 30, 40, and 50 degrees C. Then, d ln [eta]/d(l/T) and the Mark-Houwink exponents were calculated as the indices for chain flexibility and molecule conformation, respectively. These results showed: the intrinsic viscosities decreased linearly with increasing temperature, therefore, a temperature-induced conformational transition did not occur for all 10 different molecular weight chitosans in the temperature range studied. Values of d In [eta]/d(l/T) were between 633 and 1334 and increased with decreasing molecular weight, indicating that higher molecular weight chitosans are more flexible. Between 10 degrees and 50 degrees C, the Mark-Houwink exponents ranged 0.64-0.76 and increased with increasing temperature, indicating that the conformation of these chitosans were all in random coil, and a temperature induced conformational transition did not occur. The a* and a** Mark-Houwink exponents represent those chitosans whose molecular weights are larger and smaller than 223 kDa, respectively, and were obtained by using 223 kDa as the break point in the double logarithmic plots of the intrinsic viscosities and weight average molecular weight. Values of a** were between 0.41 and 0.54, while the a* values were from 0.96 to 1.07. These values for a** and a* indicate that larger and smaller molecular weight chitosans were in random coil and rod shape, respectively. PMID- 9730169 TI - Hydration properties of xylitol: computer simulation. AB - We present a molecular dynamics simulation of xylitol in SPC/E water using classical Gibbs ensemble molecular dynamics simulation. The simulation is done both with and without periodic charge update, and no qualitative difference in the results obtained by both methods is found. The analysis of the radial and angular distribution functions, the water-water hydrogen bond distributions, and water residence times allow the conclusion that there is a relatively strong hydration of xylitol. This polyol adopts a single linear conformation and, from the point of view of the hydration dynamics, it should be classified as positively hydrated. PMID- 9730170 TI - Activated protein C resistance, factor V Leiden, and retinal vessel occlusion. PMID- 9730171 TI - Two types of initial ocular manifestations in intraocular-central nervous system lymphoma. AB - PURPOSE: To study initial ocular manifestations of ocular-central nervous system (CNS) lymphoma. METHODS: The authors reviewed medical records of 10 consecutive patients with intraocular-CNS lymphoma seen at Okayama University Hospital during 16 years from 1981 to 1996. RESULTS: Three patients showed only vitreous opacity as an initial sign, whereas five other patients had typical yellowish-white infiltrates at the sub-retinal pigment epithelial (sub-RPE) level without vitreous opacity. Both manifestations were found in two patients. In seven patients, ocular symptoms developed first, followed by brain lesions. In contrast, in three patients the initial presentation was brain tumor, for which they received chemotherapy; subsequently, vitreous opacity without sub-RPE infiltrates developed. The diagnosis was made by vitrectomy in four patients, three of whom had also undergone brain biopsy; by aqueous tap in one; and by brain biopsy in one. The other four patients were diagnosed clinically, and one of them was confirmed later to have lymphoma by autopsy. CONCLUSIONS: The initial ocular manifestations of intraocular-CNS lymphoma were of two types, sub-RPE infiltrates and vitreous opacity. Keeping these two manifestations in mind will help physicians consider a diagnosis of intraocular-CNS lymphoma earlier. PMID- 9730172 TI - Factor V Leiden, activated protein C resistance, and retinal vein occlusion. AB - BACKGROUND: Resistance to activated protein C (APC resistance) is a thrombophilic abnormality characterized by a normal plasma level of protein C and an inherited defect in the coagulative response. This condition is believed to be caused by a point mutation in factor V, the so-called factor V Leiden, and is inherited as an autosomal dominant trait. PURPOSE: A case-control study was carried out to evaluate the prevalence of APC resistance and factor V Leiden in patients with retinal vein occlusion (RVO) and in control subjects. METHODS: Eighty-four consecutive RVO patients and 70 controls were tested for APC resistance with a commercial assay (Chromogenix). The first 30 patients and 47 controls were also studied for factor V Leiden. In addition, a repeat APC-resistance test was performed in 40 RVO patients and in 9 controls with a second-generation assay done to compare the reliability and reproducibility of the tests. RESULTS: Results of testing for APC resistance with the first-generation assay revealed positive results in 38 (45%) of the study patients and 6 (9%) of the controls. The difference in frequencies of APC resistance in patients and controls was statistically significant (P < 0.0001). In the patients tested for factor V Leiden, one (3%) was a heterozygous carrier of the Arg506GIn mutation and one (2%) of the controls was a heterozygous carrier. No homozygous individuals were identified in either the study or the control groups. The difference in frequencies of factor V Leiden in study patients and controls was not statistically significant (P = 1). The repeat APC-resistance assay using factor V deficient plasma in 40 RVO patients and 9 controls did not show any significant difference between study patients and controls or an association between APC resistance and the determination of the factor V Leiden mutant. CONCLUSION: The first-generation commercial assay for APC resistance is not a useful screening test. The molecular test for factor V Leiden is the only definitive method. Furthermore, no significant association was found between factor V Leiden and retinal vein occlusion. Accordingly, routine testing for the presence of the factor V Leiden mutant is not advisable for patients with retinal vein occlusion. PMID- 9730173 TI - Epiretinal membranes surrounding idiopathic macular holes. AB - PURPOSE: To describe the features of the epiretinal membranes (ERMs) surrounding idiopathic macular holes. METHODS: The charts of 83 consecutive patients (85 eyes) who underwent macular hole surgery with a systematic search for an ERM around the hole were reviewed. Visual acuity testing, fundus biomicroscopy, red free and blue filter fundus photographs, and fluorescein angiograms were performed before and after surgery. Eyes with and without ERM removal were compared. RESULTS: An ERM was removed from 26 of 85 eyes (30.6%). ERMs were found more frequently in stage 4 than in stage 3 macular holes (76.9% versus 24.6%; P < 0.01). Holes had been present for longer in eyes with ERM than in those without (14.7 versus 8.6 months; P = 0.05). Fewer stage 3 holes with an ERM had an operculum than those without (P = 0.01). The outcome and complication rates were similar in eyes with an ERM and those without. Of the 24 ERMs detectable on blue filter fundus photographs, only 11 (45.6%) were visible on red free photographs. CONCLUSION: We support the hypothesis that the presence of ERMs surrounding idiopathic macular holes is secondary to hole formation. PMID- 9730174 TI - Macular hole surgery using thrombin-activated fibrinogen and selective removal of the internal limiting membrane. AB - PURPOSE: To evaluate a tissue sealant (autologous cryoprecipitate activated with bovine thrombin) as an adjuvant in macular hole surgery. METHODS: Sixty-nine patients with stage 2, 3, or 4 full-thickness macular hole were enrolled consecutively in a prospective pilot study. Anatomic closure of the macular holes with a single operation was the primary outcome. Fifty-eight patients had pre- and postoperative standardized measurements including best refracted visual acuity, reading speed, and contrast sensitivity. Group A patients (45) had primary macular holes; Group B patients (13) had recurrent macular holes or macular holes with "other" retinal pathology. Surgical technique was standardized and membrane dissections were optional. RESULTS: The anatomic closure rate was 80% with a minimum of 6 months follow-up. Mean improvement in visual acuity for Group A (2.9+/-0.4 lines) was significantly better than for Group B (0.8+/-0.5 lines; P = 0.008). Eyes that underwent internal limiting membrane (ILM) dissections had an anatomic closure rate of 96% (23/24), compared with 71% (32/45) in "non-ILM" cases (P = 0.034). Adverse reactions included sterile hypopyon (10%), intraretinal hemorrhage (9%), pigmentary hyperplasia (3%), and retinal detachment (3%). CONCLUSION: Tissue sealants should be evaluated as an adjuvant in macular hole surgery in a randomized clinical trial. Inflammatory reactions may occur in some patients. Internal limiting membrane dissection may improve anatomic closure rates without adversely affecting the visual acuity. PMID- 9730175 TI - Repositioning dislocated posterior chamber intraocular lenses. AB - PURPOSE: Removing a dislocated posterior chamber intraocular lens (IOL) is a hazardous procedure that may lead to severe complications and visual loss. A new technique for handling this complication by means of vitrectomy to help replace the luxated IOL directly in the anterior chamber is described. METHODS: Five patients (two men and three women) underwent surgery for replacement of a luxated posterior chamber IOL in the vitreous cavity. The average age was 67.4+/-8.9 (years+/-SD); range, 57-79 years. Before and after vitrectomy, specular microscopy study of the corneal endothelium was done. RESULTS: The results obtained in these five patients were highly satisfactory, with an average final visual acuity of 20/32 and residual refraction of -1.125+/-1.33. There was no evidence of corneal endothelium decompensation. In one patient, a retinal detachment occurred after 7 months, which was operated successfully. CONCLUSIONS: This technique-which allows permanent, stable, and controllable replacement of an IOL with minimal trauma to the iris and corneal endothelium-avoids postoperative induced astigmatism. The technique is useful for IOL with and without positioning holes. No corneal incision is needed; the standard sclerotomies for vitrectomy are used. PMID- 9730176 TI - Good visual acuity in an adult population with marked posterior segment changes secondary to retinopathy of prematurity. AB - PURPOSE: To show that good visual acuity can be compatible with marked posterior segment changes secondary to retinopathy of prematurity (ROP) in the adult population. METHODS: A retrospective chart review of adult patients with regressed ROP. We found 14 eyes in 12 patients who were older than 21 years with a visual acuity of 20/60 or better associated with marked posterior segment changes secondary to ROP. RESULTS: Of the 14 eyes, 11 were myopic, with six eyes having the spherical equivalent of > or = -6.00. Best-corrected visual acuities ranged from 20/15-20/60. One eye had a macular fold. Thirteen eyes had macular ectopia. CONCLUSIONS: Good vision can be compatible with marked posterior segment changes secondary to ROP in an adult population. This emphasizes the need to follow these patients closely during childhood and treat them promptly for any amblyogenic condition that could prevent them from reaching their full visual potential. PMID- 9730177 TI - Intraocular penetration of gentamicin after once-daily aminoglycoside dosing. AB - BACKGROUND: Intraocular concentrations-particularly intravitreal concentrations after systemic administration of gentamicin are poor. Once-daily aminoglycoside dosing of intravenous gentamicin achieves peak serum levels up to five times higher than conventional dosing. Whether these increased serum levels of gentamicin improve the aqueous or vitreous concentrations in humans has not been determined. The authors sought to determine if the intraocular penetration of gentamicin would be improved using this method. METHODS: Patients undergoing vitrectomy procedures were administered intravenous gentamicin in a dose of 7 mg/kg approximately 1 hour before surgery. An adjustment in dosing was made for anyone more than 20% over his or her ideal body weight. Aqueous, vitreous, and serum samples were collected before any intraocular surgical manipulation. The samples were analyzed by fluorescence polarization immunoassay (TDx system). RESULTS: The average single gentamicin dose was 498 mg (range, 360-700 mg). The aqueous, vitreous, and serum levels averaged 1.14 microg/mL, 0.41 microg/mL, and 22.07 microg/mL, respectively. No correlation between serum level concentrations and time of administration was found for the aqueous and vitreous levels in this study. CONCLUSION: Although the average peak serum level of gentamicin was five times higher than previously reported, the vitreous levels averaged only 1.5 times higher. The blood-retinal barrier is difficult to penetrate even when higher serum levels are achieved. Due to its poor ocular penetration, gentamicin may not be among the best drugs for prophylaxis of penetrating eye injuries, surgical prophylaxis, or treatment of endophthalmitis. PMID- 9730178 TI - Varix of the vortex vein ampulla simulating choroidal melanoma: report of four cases. AB - BACKGROUND: Varix of the vortex vein ampulla is a condition that can cause diagnostic confusion with choroidal melanoma. METHODS: A case series review was performed from the Ocular Oncology Service, Wills Eye Hospital. RESULTS: In all four cases, the patients were referred with the diagnosis of a small choroidal melanoma. The lesions were located in the nasal quadrant of the fundus near the equator. One patient had two lesions in the same quadrant. In all cases, the fundus lesion became more prominent when the eye gazed in the direction of the lesion and diminished in primary gaze. The mass measured up to 6.0 mm in base diameter and 2.5 mm in thickness in proper gaze. B-scan ultrasonography showed acoustic solidity and gaze-evoked dynamic enlargement of the lesion. Indocyanine green angiography demonstrated early pooling of dye and gaze-evoked fluctuation of the hyperfluorescence in the lesion. Color Doppler imaging, performed in one patient, showed a vascular lesion of venous origin that filled when the eye was placed in the direction of the lesion. CONCLUSIONS: Varix of the vortex vein is a condition that should be considered in the differential diagnosis of equatorial small choroidal melanoma. The dynamic nature of the lesion is characteristic and diagnostic. PMID- 9730179 TI - Bartonella serology for patients with intraocular inflammatory disease. AB - PURPOSE: To determine the role of Bartonella henselae in intraocular inflammatory disease and identify its clinical features. METHODS: We retrospectively determined the serum immunoglobulin (Ig)G and IgM antibodies against B. henselae and Bartonella quintana by enzyme immunoassays in stored sera of 138 consecutive newly referred patients with uveitis who, during the acute stage of their ocular disease, underwent a standardized screening protocol to determine the cause of uveitis. RESULTS: For the entire series, the frequency of high positive levels of IgG (above 1:900) or IgM (above 1:300) antibody against B. henselae was 6% (8/138) and 3% (4/138), respectively. Except for cross-reactions between B. henselae and B. quintana, we did not find additional evidence for cross-reactions among the various bacteria tested (Coxiella burnetii and Chlamydia pneumoniae). All patients with proven infectious uveitis (n = 21) and those with established uveitic entities (n = 37) had negative B. henselae serology. High positive IgG levels were observed in 9% of patients (5/54) with unknown cause of uveitis, in two subjects with human leukocyte antigen (HLA)-B27 positive uveitis, and in one with sarcoidosis. Five patients with uveitis of unknown origin and highly elevated IgG levels against B. henselae exhibited clinical features characterized by papillitis with surrounding retinal focal lesions or edema. CONCLUSIONS: The serologic and clinical data indicate that uveitis in seropositive cases may be caused by B. henselae. We do not recommend including testing for B. henselae in initial screening of patients with uveitis, but consider it worthwhile for those with papillitis and screening results within normal limits. PMID- 9730180 TI - A comparison of self-reported utilization of ophthalmic care for diabetes in managed care versus fee-for-service. AB - PURPOSE: To assess the association between structural factors in the health care delivery system and self-reported utilization of ophthalmic services by patients with diabetes in the Medical Outcomes Study (MOS). METHODS: Self-reported utilization of ophthalmic services within the 6 months preceding enrollment into the MOS among 522 of 567 individuals with diabetes in the MOS longitudinal panel was measured. Use of eye care services was regressed (logistic model) on patient demographics, geographic location, physician specialty, type of practice, and finance plan (prepaid or fee-for-service). RESULTS: None of the variables was significantly associated with a higher or lower likelihood of having used ophthalmic services in the preceding 6 months. Thus, no difference between prepaid or fee-for-service plans or among solo practice, large multispecialty groups, or HMOs were identified. Having seen an internist, family practitioner, or diabetes specialist for diabetes care was not related to use of ophthalmic services. CONCLUSIONS: Despite a presumed greater interest in preventive health, prepaid health plans were no more or less likely than the fee-for-service sector to have patients with diabetes reporting an eye examination within the prior 6 months. Thus, steps to improve the rate of eye examinations of diabetics may need to focus beyond the structural elements of the health care delivery system. PMID- 9730181 TI - Legal blindness and employment in patients with juvenile-onset macular dystrophies or achromatopsia. AB - PURPOSE: The purpose of this study was to gain information about the employment status of legally blind patients. METHODS: Fifty-two patients with one of four juvenile-onset macular dystrophies or achromatopsia responded to questions about their employment histories and their psychological well-being. Results from the questionnaire were analyzed using z-tests for differences in proportions or t tests for differences in means. RESULTS: Forty-eight percent of the patients reported that they were employed and 52% that they were not employed. The subgroup that was not employed had a significantly higher proportion of women than men, whereas the employed group had approximately equivalent proportions of men and women. The employed subgroup reported that their success at work was due to social support. This subgroup had significantly higher household incomes, was significantly less likely to collect disability-income benefits, had significantly higher educational levels, had significantly higher positive affect, and had significantly lower negative affect than the subgroup that was not employed. A logistic regression analysis indicated that education was the primary predictor of employment. CONCLUSION: Analysis supports the conclusion that it is beneficial for legally blind individuals to obtain an optimal level of education and receive suitable social support to facilitate their successful employment. PMID- 9730182 TI - Establishment of a rabbit model of extrascleral extension of ocular melanoma. AB - PURPOSE: To establish an animal model of extrascleral extension of choroidal melanoma. METHODS: Pigmented choroidal tumors were established in nine New Zealand albino rabbit eyes using B16F10 melanoma cell line. The sclerotomy site was not closed in the subgroup of six rabbits where extrascleral extension was desired. For the control group, the sclerotomy site was sutured with 8-0 nylon. Animals were treated with daily injections of cyclosporine and followed by serial fundus examinations, color Doppler imaging, and fundus photography. All tumor bearing eyes were enucleated at the end of the follow-up period and examined for extrascleral extension. RESULTS: Extrascleral extension of choroidal melanoma occurred in all six animals with open sclerotomy sites. No extrascleral extension was observed in the control group. Color Doppler imaging identified extrascleral extension which was confirmed on gross histology. CONCLUSIONS: Our animal model of extrascleral extension of choroidal melanoma requires minimal surgery to establish, and is reproducible and easy to follow with standard diagnostic equipment. PMID- 9730183 TI - Diagnostic and therapeutic challenges. Lesions of the optic nerve head. PMID- 9730184 TI - Arterial vascular occlusion associated with factor V Leiden gene mutation. PMID- 9730185 TI - Bilateral retinal vein occlusion associated with factor V Leiden mutation. PMID- 9730186 TI - Branch retinal vein occlusion after spontaneous obliteration of retinal arterial macroaneurysm. PMID- 9730187 TI - Bilateral choristomas in the ocular adnexa, lids, and posterior segment. PMID- 9730188 TI - Chorioretinopathy in primary antiphospholipid syndrome: a case report. PMID- 9730189 TI - Multiple bilateral choroidal metastatic tumor from bronchial carcinoid: case report. PMID- 9730190 TI - Radiation therapy for exudative age-related macular degeneration. PMID- 9730191 TI - Peripheral transscleral retinal diode laser for rubeosis iridis. PMID- 9730193 TI - Nutrient intakes and growth of very low birth weight infants. AB - OBJECTIVE: Our purpose was to determine nutrient intakes and growth of very low birth weight (VLBW) infants. STUDY DESIGN: The survey consisted of infants admitted during a 9-month period to a tertiary neonatal center. Data were obtained concerning all 51 infants born weighing <1300 gm who survived beyond 21 days of age. METHODS: At weekly intervals, intakes of fluid, energy, and protein from all sources were determined and body weight was recorded. RESULTS: During the first 2 weeks of life, intake of energy (predominantly parenteral) averaged 75 +/- 12 kcal/kg per day and intake of protein averaged 1.9 +/- 0.5 gm/kg per day. From 15 to 35 days, intake of energy (transition from parenteral to enteral) averaged 99 +/- 12 kcal/kg per day and intake of protein averaged 2.5 gm/kg per day. During the period 36 to 56 days (early enteral) and 57 days to term (late enteral), energy intakes were 108 +/- 13 and 110 +/- 15 kcal/kg per day, respectively, and protein intakes were 2.7 +/- 0.5 and 2.7 +/- 0.5 gm/kg per day, respectively. These low intakes of energy and protein (relative to presumed requirements) were explained by low intake volumes and low protein concentrations of feedings. Weight reached birth weight by 14 days of age. Subsequently, weight gains averaged 13.0, 13.8, and 11.6 gm/kg per day, respectively, in successive periods. These gains were lower than would have occurred in utero. CONCLUSION: Observed growth of VLBW infants was slow relative to in utero growth, presumably because intakes of energy and, in particular, of protein fell short of intakes needed to duplicate in utero growth. Changes in feeding practices, as well as in composition of feedings, are needed if in utero growth is to be matched. PMID- 9730194 TI - The value of the Infant/Child Monitoring Questionnaire for identification of developmental disability among neonatal intensive care unit graduates. AB - OBJECTIVE: We hypothesized that the Infant/Child Monitoring Questionnaire (ICMQ) could be used to identify at-risk infants eligible for developmental interventional services. STUDY DESIGN: Of this cross-sectional observational study, group A (n = 108) included a retrospective review of moderate risk infants scheduled for developmental assessment clinic (DAC) visits. Group B (n = 108) included moderate-risk infants whose parents completed the ICMQ. Group C (n = 67) included high-risk infants who were seen in the DAC and whose parents completed the ICMQ. RESULTS: For group A infants, 43.5% were seen in the DAC; 10.6% of these visits resulted in an intervention. For group B infants, 56.5% of parents completed the ICMQ; 66.7% of subsequent visits resulted in an intervention. For group C infants, comparison of ICMQ and DAC visits showed moderate agreement (kappa = 0.50). CONCLUSION: The ICMQ is a useful tool to identify moderate-risk infants requiring further intervention, but caution must be used when applied to high-risk infants. PMID- 9730195 TI - Very low birth weight infants and their families during the first year of life: comparisons of psychosocial outcomes based on after-care services. AB - OBJECTIVE: To evaluate psychosocial outcomes in families of very low birth weight infants during the first year postdischarge. STUDY DESIGN: This was a prospective investigation of family functioning in families of 81 very low birth weight infants discharged from two tertiary care neonatal intensive care units in Los Angeles, CA. Infants were assigned to four groups receiving a variety of after care services in their homes. Analyses of variance and t-tests were used to examine differences in outcomes, including parental involvement with infant, maternal depression, and family adaptation and cohesion over time. RESULTS: During the first year following discharge, there were no differences between after-care groups in levels of maternal depression as measured by the Center for Epidemiologic Studies Depression Scale or family cohesion and adaptability as measured by the Family Adaptability and Cohesion Evaluation Scales II. Significant between-group differences were seen on measures of home environment at both 6 and 12 months and self-reports of satisfaction with parenting at 6 months. CONCLUSION: Results of this study indicate that types of after-care services do not change basic maternal or family characteristics. However, long term home-visiting services appear to impact the way mothers interact with their high-risk infants. Furthermore, such services seem to influence a mother's perception of satisfaction with her role. PMID- 9730196 TI - The timing of skin acidification in very low birth weight infants. AB - OBJECTIVE: The purpose of this study was to examine the development of the pH mantle of the skin in very low birth weight (VLBW) infants. STUDY DESIGN: Forty VLBW infants underwent repeated measurements of skin pH over the first month of life using a glass flat-surface pH electrode. Six skin sites were measured, daily for the first week of life and then twice weekly for the next 3 weeks. RESULTS: The only factor that affected the initial skin pH was sex, with males having a significantly higher pH at birth. Over time, birth weight, skin area, and postnatal age had significant effects on skin pH. The pattern of postnatal change in skin pH was similar to that described in term infants, a rapid decrease in pH over the first week followed by a more gradual decrease over the next 3 weeks. CONCLUSION: The development of the skin's acid mantle in VLBW infants occurs rapidly during postnatal life and closely mimics the pattern seen at term. PMID- 9730197 TI - Meta-analyses of surfactant replacement therapy of infants with birth weights less than 2000 grams. AB - OBJECTIVES: We conducted a meta-analysis of surfactant replacement therapy to determine (1) the efficacy of surfactant therapy in the reduction of short-term morbidity and long-term outcome in terms of bronchopulmonary dysplasia (BPD) and mortality; (2) whether there are differences in efficacy between modified natural surfactant and synthetic surfactant; (3) the effectiveness of prophylactic surfactant therapy; and (4) whether there are differences in efficacy between the prophylactic approach and the rescue strategy. STUDY DESIGN: We included studies in which infants with birth weights between 500 and 1500 gm were eligible. Studies were grouped into the following categories: (1) rescue therapy with modified natural surfactant; (2) rescue therapy with synthetic surfactant; (3) prophylaxis with modified natural surfactant; (4) prophylaxis with synthetic surfactant; (5) prophylaxis versus rescue studies; (6) modified natural surfactant versus Exosurf (Burroughs-Wellcome Co., Research Triangle Park, NC) studies. The relative risk ratios, corrected for study size, were calculated for the outcome variables (pneumothorax, incidence of BPD, survival, survival without BPD, prevention of hyaline membrane disease [HMD], and intraventricular hemorrhage [IVH]). RESULTS AND CONCLUSION: Surfactant therapy is efficacious in reducing the risk for pneumothorax and increasing the chance for survival without BPD. Synthetic surfactant is not efficacious in the prevention of HMD. Modified natural surfactant is more effective in reducing the risk of pneumothorax and increasing the chance for survival without BPD than is synthetic surfactant. These data do not support the use of either synthetic or modified natural surfactant for routine prophylaxis. PMID- 9730198 TI - Intra-amniotic pressure reduction in twin-twin transfusion syndrome. AB - OBJECTIVE: Our purpose was to measure intra-amniotic pressure before and after decompression amniocentesis in twin-twin transfusion syndrome. STUDY DESIGN: Intra-amniotic pressures were measured during decompression amniocentesis on 18 occasions in 5 pregnancies complicated by twin-twin transfusion syndrome. The intra-amniotic pressure was determined with a water manometer before and after removal of amniotic fluid. For comparison, intra-amniotic pressure was determined in 10 uncomplicated gestations. RESULTS: Initial intra-amniotic pressures in twin twin transfusion gestations (mean, 17.2 +/- 5.2 cm H2O; range, 5.5 to 33.0 cm H2O) were higher than those of the uncomplicated gestations (mean, 8.4 +/- 3.3 cm H2O; range, 3.5 to 13.5 cm H2O; p < 0.002). Intra-amniotic pressures following therapeutic amniocentesis (mean, 10.9 +/- 5.1 cm H2O; range, 3.5 to 23.0 cm H2O) were not different from those of the uncomplicated gestations (p = 0.16). CONCLUSION: The intra-amniotic pressure in twin-twin transfusion gestations is higher than that of the uncomplicated gestation. Decompression amniocentesis reduces intra-amniotic pressure to that of the uncomplicated gestation. PMID- 9730199 TI - Legionella pneumonia in neonates: a literature review. AB - It has recently been recognized that neonates may develop pneumonia as a result of Legionella pneumophila. The objective of this study is to characterize the epidemiology, risk factors, diagnosis, clinical features, and outcome of neonatal legionellosis. Review of the literature revealed nine cases of neonatal Legionella infection. Five neonates were term infants and four were preterm. Eight had potential risk factors such as prematurity, congenital heart disease, bronchopulmonary dysplasia, or corticosteroid therapy. Diagnosis was proven by culture in all cases. The main presentation was acute respiratory distress requiring mechanical ventilation. In six infants, the infection had a fatal outcome, including five who were not treated with erythromycin. All the cases were nosocomial, and environmental Legionella was documented in five cases. As has been noted in adults and children with Legionella, early recognition and institution of appropriate therapy are the most important determinants of the prognosis. PMID- 9730200 TI - Radiological pulmonary changes during gram-negative bacillary nosocomial bloodstream infection in premature infants. AB - OBJECTIVE: The objective of the study was to characterize the changes that occur in chest radiographs at the time of gram-negative bacilli (GNB) nosocomial bloodstream infection (BSI) and to determine the contribution of bronchopulmonary dysplasia (BPD) and airway gram-negative bacterial pathogens to the clinical diagnosis of GNB nosocomial pneumonia. STUDY DESIGN: This retrospective investigation involved 41 BSI infants (study group) and 50 GNB airway colonized infants who had sepsis workup with negative blood cultures (control group). We correlated clinical findings (95 blood and 305 tracheal aspirate (TA) cultures) with radiographic findings noted within 2 days before, the day of, and the day after blood cultures. Two radiologists independently graded 258 films using a modified score for BPD and a semiquantitative score ("probable," "possible," or "unlikely") for pneumonia. RESULTS: Mean birth weight was 1057 gm and 1044 gm for the study and control groups, respectively. Of the study population, 54% were male, 21% were black, 89% received surfactant, 79% received dexamethasone, and 88% survived. Average age at the time of blood cultures for both groups was 23 days. Most common isolates from blood and TA were Klebsiella pneumoniae, Enterobacter cloacae, Escherichia coli, and Pseudomonas aeruginosa. Eight BSI infants died, mainly as a result of P. aeruginosa and K. pneumoniae; three control patients also died. Radiological findings were that BPD scores did not change in relation to BSI and were similar between study and control groups. Of the BSI patients, 21 presented with newly positive TA at the time of positive blood culture; "probable" or "possible" pneumonia was diagnosed in 18 of them. Their BPD scores were higher than those of the remaining BSI patients, of whom seven were already airway colonized, nine had negative TAs, and four were not intubated. Only one of these 20 patients had "possible" pneumonia noted on chest x-ray films. CONCLUSION: Radiographic signs of air space disease accompanied by the recovery of GNB respiratory pathogens from the blood and from a previously uncolonized airway strongly support the clinical diagnosis of GNB nosocomial pneumonia. Radiological signs of BPD are stable in relation to nosocomial BSI caused by GNB, but BPD radiological scores are higher among infants who also had a newly acquired respiratory GNB. BSI, new respiratory tract GNB, and BPD are critical associations for the clinical interpretation of radiographic changes in the ventilated newborn. PMID- 9730201 TI - Perinatal outcome and the social contract--interrelationships between health and humanity. AB - Rates of infant mortality and prematurity or low birth weight serve as indirect measures of the health of a nation. This paper presents current population data documenting the still serious problem of perinatal outcome in the United States as well as in other economically developed countries. International comparisons suggest that nations with the greatest inequality of income and social opportunity also have the most adverse perinatal, child, and adult health outcomes. Furthermore, the data assert that these effects are independent of average national wealth or gross national economic productivity. Health status differs by social class and race, even among the most affluent sectors of the population. All social classes, even the wealthiest, suffer the health consequences of social inequalities. An explanatory sociopsychologic theory of causality is proposed. PMID- 9730202 TI - Resuscitation of a micropremie: the case of MacDonald v. Milleville. AB - One of the most unsettling experiences for a neonatologist is having an early gestational-age infant for whom resuscitation has been abandoned or not initiated subsequently begin breathing on his own. That was the experience of Gregory Milleville, MD when at 2:30 AM a nurse brought such an infant with a heart rate of 130 and a temperature of 91.2 degrees F to the neonatal intensive care unit (NICU). PMID- 9730203 TI - Fulminant necrotizing enterocolitis in a premature neonate treated for supraventricular tachycardia. AB - A premature neonate with supraventricular tachycardia was treated prenatally and postnatally, without significant signs of congestive heart failure. Enteral feeding was initiated after 48 hours of age. The infant developed fatal, fulminant necrotizing enterocolitis 28 hours after starting feeds. PMID- 9730204 TI - Spontaneous resolution of nonimmune hydrops in a fetus with a cystic adenomatoid malformation. AB - Congenital cystic adenomatoid malformation (CCAM) is a rare pulmonary anomaly. It is a hamartomatous lesion characterized by a cessation of normal bronchiolar maturation, resulting in overgrowth of the terminal bronchioles. There is no preference for sex or location, and usually the lesion is confined to a single lobe. CCAMs have been classified into three subtypes according to the presence of and size of the cysts. Type I lesions have large cysts (2 to 10 cm in diameter), type II have smaller cysts (< 1 cm in diameter), and type III is noncystic. There have been several reports of diminution in size of these lesions and complete regression. However, in those cases fetal hydrops was absent. In the presence of nonimmune hydrops, fetal prognosis is extremely poor without any intervention. There are only two case reports describing fetal survival without intervention when nonimmune hydrops is present. We present a case of survival of a fetus with CCAM and nonimmune hydrops diagnosed at 24 weeks' gestation. PMID- 9730205 TI - Congenital candidiasis: varied presentations. AB - Congenital candidiasis, especially the disseminated disease, is very uncommon but has been reported in very low birth weight infants. Five cases of congenital candidiasis, two with cutaneous type and three with systemic type, are described. All cases were symptomatic within the first 24 hours of life and none of them had significant risk factors such as the presence of foreign body in the maternal genital tract. Cutaneous candidiasis presented as extensive erythematous rash with infiltrative plaques in one and as bullous lesions in the other. Three infants who had disseminated candidiasis presented with extreme leukemoid reaction, severe hyperglycemia, and skin mottling with some patchy areas resembling first-degree burns, respectively. One infant had meningitis and the autopsy of another who died revealed several microabscesses containing Candida spores in the liver and lungs. The urine microscopy obtained by suprapubic bladder aspiration was found to be a good diagnostic marker of systemic invasion. The purpose of this report is to highlight the importance of recognizing candida as a possible pathogen in a critically ill neonate even though the clinical presentation may be nonspecific and varied. The presence of characteristic skin lesions of Candida species within 24 hours of life is an important clue to the possible diagnosis of congenital candidal infection. Even though very high mortality has been reported in congenital disseminated candidiasis, early recognition and treatment could give a favorable outcome. PMID- 9730206 TI - Treatment of obstructive jaundice in erythroblastosis fetalis with ursodeoxycholic acid (UDCA): a case report. AB - OBJECTIVE: To report a significant improvement of direct hyperbilirubinemia values, in an infant with cholestasis secondary to erythroblastosis fetalis, after treatment with ursodeoxycholic acid (UDCA). STUDY DESIGN: Case report. RESULTS: A full term infant, with total and direct bilirubin values of 26 mg/dl (445 micromol/l) and 24.5 mg/dl (419 micromol/l), respectively, on the third day of life, had total and direct bilirubin values of 8.2 mg/dl (140 micromol/l) and 6.9 mg/dl (118 micromol/l), respectively, after 2 days of treatment with UDCA. Because the natural course of this cholestasis takes several weeks to resolve, the observed improvement is highly suggestive of a direct effect of UDCA on the disease course. CONCLUSION: This treatment may add a new therapeutic option to the limited measures available for this condition, although further studies regarding safety and its mechanism of action are needed before it can be routinely recommended. PMID- 9730207 TI - Neonatology radiology casebook. Tracheal agenesis. PMID- 9730208 TI - Special imaging casebook. CHARGE association-DiGeorge syndrome with congenital short esophagus and single kidney. PMID- 9730209 TI - Response to "Induced early delivery of a fetus with hypophastic left heart: a moral choice when neither surgery nor abortion is an acceptable option". PMID- 9730210 TI - Resting and nesting in primates: behavioral ecology of inactivity. PMID- 9730211 TI - Nests, tree holes, and the evolution of primate life histories. AB - In contrast to the majority of primates, many prosimians, some New World monkeys, and the great apes rest in tree holes or self-constructed nests during their inactive periods. The goal of this comparative study was to examine possible functions of this interspecific variation. Information on resting behavior, maternal behavior, and basic life-history traits was gleaned from the literature and mapped onto a phylogenetic tree of primates for various comparative tests. Parsimony-based reconstructions revealed that only the use of nests or tree holes as shelters for young infants can be unequivocally reconstructed for various higher taxa, suggesting that it is functionally different from the use of shelters by adults (who may be accompanied by infants). Further reconstructions revealed that the ancestral primate was most likely nocturnal and solitary and produced a single infant that was initially left in a shelter and later carried orally to a parking place in the vegetation--a combination of traits exhibited by many living galagos. Evolutionary losses of the use of nests were concentrated among diurnal and nonsolitary taxa and weakly associated with evolutionary increases in body size. Thus, protective functions of nests or tree holes used by prosimians are either secondary or there are alternative ways of obtaining protection. Because the evolution of larger litters was significantly associated with the presence of shelters, the presence of relatively altricial young among prosimians best explains the use of nests and tree holes, which are in most but not all cases also used by adults. These shelters therefore play an integral part in the life-history strategies of primitive primates and their ancestors and evolved secondarily among anthropoids for other purposes. PMID- 9730212 TI - Sleeping sites, sleeping places, and presleep behavior of gibbons (Hylobates lar). AB - The sleeping habits of wild white-handed gibbons (Hylobates lar) were investigated to assess the risk of predation and predation-avoidance behavior. Sleeping sites were distributed throughout home ranges, including areas where they overlapped with neighbors, and appeared to be selected independently of habitat characteristics. Individuals did not build night nests or otherwise manipulate the vegetation around the sleeping place but slept on open branches. Group members usually slept in separate trees, and, except for females with infants, they never shared a sleeping place. Sleeping trees were entered several hours before dusk and were used for about 14-17 h. The majority of sleeping trees were used only once, and fewer were selected repeatedly by the same or other group members. Usually females with infants went into a sleeping tree first, then juveniles, and last were mostly subadult and adult males. Intragroup competition over access to a sleeping place was observed once. Average time difference between the first and last group member to enter a sleeping tree was 13 min. The sequence of departure from sleeping trees was more variable. Gibbon sleeping habits seem to primarily reflect adaptations to minimize predation risk. The predation-risk hypothesis was indirectly supported by observations of mobbing pythons, alarm calls given in response to birdes of prey flying low over the canopy, and more importantly by 1) the predominant use of large sleeping trees, which were among the tallest trees available, particularly by adult females with small infants and juveniles, 2) an unpredictable long-term pattern of reuse of sleeping places, and 3) inconspicuous presleep behavior. PMID- 9730213 TI - Sleep, sleeping sites, and sleep-related activities: awakening to their significance. AB - Since primates spend about half of their life at sleeping sites, knowledge of behavior in the vicinity of sleeping sites and analysis of factors influencing their use is important for understanding the diversity of primates' adaptations to their environment. The present paper reviews recent progress in the ethology and ecology of sleep in diurnal monkeys and apes. Emphasis is given to the following topics: safety from predators at sleeping sites, physical comfort, social behavior, and psychophysiology of sleep. In all cases, study at the group level and at the individual level can provide insights into behavioral adaptations. As well as increasing understanding of behavior in the wild, knowledge of sleep-related behavior can be applied with a view to improving the environment for captive primates. PMID- 9730214 TI - Sex-specific usage patterns of sleeping sites in grey mouse lemurs (Microcebus murinus) in northwestern Madagascar. AB - Sleeping sites are a potentially important resource for grey mouse lemurs since they are confronted with high daily temperature fluctuations and a high predation pressure. In order to determine the existence and degree of resource competition, sleeping site characteristics, locations, and usage patterns as well as sleeping group compositions were investigated in a 3 month field study in a dry deciduous forest of northwestern Madagascar. The daily sleeping sites of females were on average better insulated and safer than those of males. Males used more sleeping sites and changed the site more often than females. During the whole study, males slept alone, whereas the females formed stable sleeping groups in on average 83.7% of the days. Sex-specific differences in usage patterns might be explained by intersexual resource competition and female dominance and could possibly be related to differential parental investment of the sexes. The underlying study indicates that sleeping sites may be a restricted and defendable resource for grey mouse lemurs. The investigation gives new insights into the distribution patterns and social organization of this species. PMID- 9730215 TI - Shadows on a changing landscape: comparing nesting patterns of hominids and chimpanzees since their last common ancestor. AB - Studying the evolution of nesting behavior within the human-chimpanzee clade is problematic because evidence is sparse and difficult to interpret. Lacking a fossil or archaeological record for proto-chimpanzees, reconstructions of the antecedents of modern chimp nesting patterns can be reconstructed only from careful studies of variation in current chimpanzee and bonobo nesting patterns within the context of spatial and temporal landscape parameters. The ethology of nesting also provides an important frame of reference for reconstructions of early hominid nesting behavior. If the contemporary contrast between human and chimpanzee nesting patterns is seen as an evolutionary dichotomy, then African prehistoric landmarks that mark the origin of this split might include bipedalism and the origins of the hominidae, the first stone tools and the origins of Homo, the developmental and behavioral adaptations of Homo ergaster, shifts in Late Acheulian settlement patterns, and the origins of anatomically modern humans and the Middle Stone Age. The issue of whether Early Stone Age archaeological sites were used for nesting is unresolved because potential markers of such behavior, such as hearths, structures, or bedding, are not unambiguously recognizable in the archaeological record until the Middle Stone Age. PMID- 9730216 TI - Occurrence of autoantibodies to cilia in lambs with a 'coughing syndrome'. AB - A respiratory disease of lambs that has been termed the 'coughing syndrome' has been observed in the mid-western region of the United States of America. Mycoplasma ovipneumoniae (M. ovipneumoniae) and Mycoplasma arginini (M. arginini) were routinely isolated from the respiratory tract of lambs with this disease. A high level of antibodies reactive with ovine cilia of the upper respiratory tract was detected in the sera from many of the lambs in affected flocks but not in sera of lambs from unaffected flocks. The reactivity of these antibodies with cilia was demonstrated by ELISA and confirmed by indirect immunofluorescent staining and western immunoblotting. These antibodies were predominantly of the IgG isotype. They were distinct from cold or warm agglutinins and could be absorbed from the sera with cilia but not with antigens of common bacterial pathogens of the sheep respiratory tract including M. ovipneumoniae, M. arginini, Pasteurella haemolytica, Pasteurella multocida or Neisseria ovis. In addition, their occurrence appeared to be independent of the specific antibodies to M. ovipneumoniae and M. arginini. Western immunoblotting indicated that the antibodies were directed primarily against an antigen with apparent molecular weight of 50 kDa. In one flock from which serial serum samples were collected from the same lambs over a 10-month period, antibodies to ovine cilia developed before the onset of the clinical disease and persisted for a period of several months until most of the lambs had apparently recovered. However, colonization of the respiratory tract of the lambs by M. ovipneumoniae preceded the production of these antibodies. Sequential serum samples taken from another flock, with no known history of this coughing, showed no such antibodies throughout the sampling period. It is suggested that an immunopathologic mechanism involving production of autoantibodies directed against a ciliary antigen of the lambs could be a contributing factor to the pathogenesis of this clinical disease. PMID- 9730217 TI - Short-termed expression of interleukin-12 during experimental BLV infection may direct disease progression to persistent lymphocytosis. AB - In this study an attempt was made to elucidate cellular response cytokine expression upon experimental bovine leukemia virus (BLV) infection in cattle. Progression of infection was monitored by BLV gp51 mRNA expression or DNA amplification by RT-PCR or PCR, respectively, to detect provirus infected cells. Antibodies to BLV were detected by an agar gel immuno-diffusion (AGID) test in 5 weeks and persistent lymphocytosis (PL+) was established in all four BLV-infected animals in 24 weeks after infection. At the initial stage of infection a strong cellular immune response was induced mediated by IL-12p40 mRNA expression. Short termed IL-12p40 expression was observed in peripheral blood mononuclear cells (PBMC) in two out of four infected animals following 1-3 weeks after infection, while viral mRNA expression was observed 2 weeks following infection. Expression of genes coding for the pro-inflammatory TNFalpha, IL-1beta and cellular response cytokines IFNgamma and IL-2 was detected beginning with the second and third week after infection in all BLV-infected animals. However, IFNgamma expression significantly decreased in 12 weeks after infection in three animals while IL-10 message initially detected 3 weeks after infection increased by 12 weeks and persisted. The observed immediate short-termed cell mediated immune response characterized by IL-12p40 and IFNgamma expression followed by an early shift to an IL-10 induced humoral response, may change the cytokine balance and direct disease progression to the PL+ stage. PMID- 9730218 TI - Effective in vivo depletion of T cell subpopulations and loss of memory cells in cattle using mouse monoclonal antibodies. AB - Conditions were established to obtain depletion of T lymphocyte subsets in lymphoid tissues of calves by injection of mouse monoclonal antibodies to T cell antigens. Adverse reactions were avoided by injecting small quantities of antibody, until target cells had disappeared from blood. Two different mechanisms appeared to be responsible for elimination of the target cells. Rapid depletion of T cells was associated with complement-binding antibody isotypes (IgG2a, IgM), suggesting a complement-mediated mechanism. Clearance of T cells after several days was observed with a non complement-binding isotype (IgG1), suggesting phagocytosis or induction of apoptosis as possible mechanisms. Clearance of the cells in peripheral blood and spleen was obtained with 10-20 mg of anti-CD4 or anti-CD8, but almost ten times as much was needed to obtain depletion of the cells in lymph nodes and Peyer's patches. Depletion lasted for 12 days for CD4 T cells and 3 weeks for CD8 T cells. Successful and lasting depletion (at least 2 weeks) was also obtained with other T cell reagents, such as anti-CD2 and anti WC1 (gamma/delta T cells). Although B lymphocytes could be removed by a complement-binding antibody, complete depletion of these cells only lasted for a few hours, probably because B cells regenerate faster than T cells. T cell function was severely inhibited when CD4+ T cells were depleted. Stimulation of T cells with foot and mouth disease viral antigen (FMDV) in vaccinated calves was non-existent after depletion. Even 2 months after restoration of normal CD4 T cell levels in blood, activity to FMDV was low. This suggested that the depleted T cells were replaced by naive cells. PMID- 9730219 TI - Increased MHC class II and CD25 expression on lymphocytes in the absence of persistent lymphocytosis in cattle experimentally infected with bovine leukemia virus. AB - We recently observed that a group of cattle experimentally infected with bovine leukemia virus (BLV) had enhanced antibody responses to recall antigens. None of the cattle in this group were classified as persistently lymphocytotic, but they did have significantly increased numbers of circulating T and B cells. In order to investigate the potential mechanisms of BLV-induced immune activation, dual color flow cytometry was used to compare the expression of MHC class II (MHC II) molecules and the inducible IL-2 receptor alpha chain, CD25, on lymphocyte subsets in freshly isolated and cultured PBMC from these same BLV-infected cattle (n=5) with that of age-matched, uninfected controls (n=3). Freshly isolated peripheral blood mononuclear cells (PBMC) from BLV-infected cattle were found to contain a significantly higher percentage of B cells that expressed MHC II molecules (p<0.01). In addition, an increased proportion of CD4+ T cells from BLV infected cattle expressed MHC II molecules after 20 h of Concanavalin A (Con A) stimulation (p<0.05), and MHC II expression was increased on both CD4+ (p<0.01) and CD8+ (p<0.05) T cells from BLV-infected cattle after 68 h in vitro, even in the absence of exogenous mitogen. Although CD25 expression was not increased on freshly isolated lymphocytes from BLV-infected cattle, an increased percentage of B cells from BLV-infected cattle expressed CD25 after 20 h of culture, either in the presence (p<0.05) or absence (p<0.01) of Con A. Thus, in addition to causing alterations in absolute numbers of circulating lymphocytes, BLV infection appears to cause a functional activation of both B and T cells, even in cattle that are non-lymphocytotic. It is likely that these BLV-induced alterations in lymphocyte activation status contributed to the previously observed enhancement of antibody responses in vivo. PMID- 9730220 TI - Effects of long-term infection with bovine immunodeficiency virus and/or bovine leukemia virus on antibody and lymphocyte proliferative responses in cattle. AB - Immune responses were examined in cattle between 3-5 years after experimental inoculation with bovine immunodeficiency virus (BIG) and/or bovine leukemia virus (BLV). Lymphocyte proliferative responses to Con A or to allogeneic lymphocytes with foreign major histocompatibility complex molecules (allo MHC) were determined by 3H-thymidine incorporation assays. Antigen-specific antibody and lymphocyte proliferative responses were measured following vaccination with tetanus toxoid (TT) and bovine herpes virus-1 (BHV-1). Lymphocytes from BIV infected cattle had significantly (p<0.05) reduced proliferative responses to Con A, but responses to allo-MHC and TT did not differ from those of uninfected controls. BIV infection also had little effect on TT-specific antibody responses in vivo. In contrast, BLV-infected cattle had significantly increased secondary antibody responses to vaccination with TT, as well as enhancement of antibody responses to BHV-1. Co-infection with BIV did not alter the BLV effect, suggesting a lack of significant interaction between the two viruses in vivo. Numbers of circulating mononuclear cells were also higher in BLV-infected cattle, which was attributable to increases in both T and B cell numbers. Unstimulated lymphocytes from BLV-infected cattle had significantly increased spontaneous uptake of 3H-thymidine in vitro. When differences in counts per minute were analyzed, lymphocytes from BLV-infected cattle had slightly increased proliferative responses to Con A, but no consistent alternations in responsiveness to allo-MHC, TT, or BHV-1. The observed increase in antibody responses to non-BLV antigens suggests that at least in clinically asymptomatic cattle, BLV infection may cause a non-specific B cell activation. PMID- 9730221 TI - Seasonal changes in the lymphoid organs of wild brown trout, Salmo trutta L: a morphometrical study. AB - A morphometrical evaluation was made of the seasonal changes affecting the numbers of lymphocytes in the thymus, spleen and pronephros of wild brown trout, Salmo trutta, while the size of the thymus and the three thymic zones were also determined. Results reveal statistically significant changes throughout the year in the number of lymphocytes in the lymphoid organs studied. The spleen and pronephros have similar annual patterns of lymphocyte distribution with high numbers in two seasons, spring and autumn, and two periods of lymphoid involution in winter and summer. The highest numbers of thymocytes occur in trout caught in May and August, and the lowest in winter. In addition to normal lymphocytes, degenerated lymphoid cells that show pale cytoplasm devoid of cell organelles, also occurred in all the lymphoid organs. A negative correlation exists between the numbers of normal lymphocytes and that of degenerated lymphoid cells. The thymic size, as well as that of the subcapsular, inner and outer thymic zones, undergo very significant changes over the year. We discuss the relevance of cell proliferation, cell migration and in situ cell death for the circannual variations observed in the cell content of trout lymphoid organs, together with the possible causes. PMID- 9730222 TI - Saliva of Rhipicephalus sanguineus tick impairs T cell proliferation and IFN gamma-induced macrophage microbicidal activity. AB - In this study, we investigated tick saliva effects on T cell proliferation, antigen presentation and IFN-gamma-induced macrophage activation, events which are associated with host immune defense mechanisms. Mice repeatedly infested with Rhipicephalus sanguineus ticks, similarly to dogs, did not develop resistance to further infestations. We determined that R. sanguineus tick saliva inhibited, in a dose-dependent manner, both Con-A and specific antigen-induced splenic T cell proliferation. Tick saliva diluted twenty times (64 microg/ml) inhibited Con-A induced and antigen-specific T cell proliferation in 83% and 69%, respectively. In addition, the inhibition of cell proliferation correlated with a decrease in IL-2 production. Microconcentrator fractionated saliva was tested on a Con-A induced cell proliferation assay, and showed that one fraction between 3 and 10 kDa and another smaller than 3 kDa can be responsible for the inhibition of T cell proliferation. Although saliva inhibited cell proliferation, it did not impair antigen presentation. Tick saliva further abrogated the killing of intracellular forms of Trypanosoma cruzi by IFN-gamma-activated macrophages. Moreover, saliva-induced macrophage inhibition of IFN-gamma-induced-trypanocidal activity was paralleled with 69% less nitric oxide (NO) production. Finally, tick saliva doubled the production of IL-10 and reduced 84.6% production of IFN-gamma by splenocytes cultured with T. cruzi, suggesting that decreased macrophage NO production may be due to a saliva-induced cytokine imbalance, leading to decreased NO synthase activity. Together, these data indicate that tick saliva can modulate host immune response, thus, contributing to its feeding success and favoring the transmission of tick-borne pathogens. PMID- 9730223 TI - A rational strategy for enhancing the affinity of vancomycin towards depsipeptide ligands. AB - Glycopeptide antibiotics with enhanced affinity for model depsipeptide ligands may also exhibit enhanced efficacy against bacteria exhibiting the vanA resistance phenotype. To design modified agents with enhanced affinity for these ligands, and better understand why traditional agents have low affinity for depsipeptide ligands, free energy perturbation studies were performed on vancomycin derivatives by means of molecular dynamics simulation. The results suggest that modifications of the asparagine side chain on residue 3 of the antibiotic which enhance its hydrophobicity will enhance the affinity of glycopeptide antibiotics for depsipeptide ligands, and act synergistically with other modifications that enhance the efficacy of these agents against vanA positive bacteria. PMID- 9730224 TI - Combinatorial synthesis and screening of a chemical library of 1,4 dihydropyridine calcium channel blockers. AB - Solid-phase synthesis of a 300-member pharmacophore library of 1,4 dihydropyridines from keto ester, diketone and aldehyde building blocks on a cleavable amine polymeric support is described. Screening and serial deconvolution of the combinatorial library has resulted in identification of known and new potent calcium channel blockers. PMID- 9730225 TI - Reaction medium engineering in enzymatic peptide fragment condensation: synthesis of eledoisin and LH-RH. AB - The influence of different reaction systems on alpha-chymotrypsin-catalyzed synthesis of eledoisin and LH-RH peptides from (7 + 4) and (5 + 5) fragments was investigated. The peptide yield was determined in the following systems: buffered aqueous media, frozen solutions, organic media, and cosolvent mixtures. The experimental set up was tailored to allow the screening of an array of conditions with minimum consumption of peptide fragments (2.1 and 2.5 mM). The best yields (22% yield for eledoisin and 68% yield for LH-RH) were obtained in buffered aqueous solutions. It was found that the choice of buffer had a strong influence on the peptide yield; boric-borate and ammonium acetate buffers at pH 9, gave the best results. In buffered aqueous systems, both syntheses were scaled up by using a 10-fold increase in fragment concentration (21 and 25 mM). Under these conditions the yields rose to 57% and 80% of eledoisin and LH-RH, respectively. Moreover, during the synthesis of eledoisin and in the presence of boric-borate buffer pH 9, the peptide precipitated from the reaction medium preventing a secondary hydrolysis and facilitating the in situ product purification. PMID- 9730226 TI - Quantitative structure-activity studies of octopaminergic agonists and antagonists against nervous system of Locusta migratoria. AB - The quantitative structure activity relationship (QSAR) of octopaminergic agonists and antagonists against the thoracic nerve cord of the migratory locust, Locusta migratoria L., was analyzed using physicochemical parameters and regression analysis. The hydrophobic effect, dipole moment, and shape index were important in terms of Ki: the more hydrophobic, the greater dipole moment, and the smaller shape index of the molecules, the greater the activity. A receptor surface model (RSM) was generated using some subset of the most active structures. Three-dimensional energetics descriptors were calculated from RSM/ligand interaction and these three-dimensional descriptors were used in QSAR analysis. This data set was studied further using molecular shape analysis. PMID- 9730227 TI - Inhibitors of glycogen phosphorylase b: synthesis, biochemical screening, and molecular modeling studies of novel analogues of hydantocidin. AB - The synthesis and biochemical screening of four novel spironucleosides 1-4 against rabbit liver glycogen phosphorylase b (Gpb), along with molecular modeling studies on compound 2 and its 4-hydroxy analogue VII, have been presented. Gpb is a key enzyme of glycogen metabolism, and is known to be involved in the control of diabetes mellitus. The general strategy for synthesis involved base-catalyzed condensation of diethyl 2,4-dioxoimidazolidine-5 phosphonate (5) with either 2-deoxy-D-ribose or D-ribose, followed by sequential reactions involving ring-closure with phenylselenenyl chloride and reduction with tri-n-butyltin hydride catalyzed by azobisisobutyronitrile. Compounds 2 and 4 were found to be weak competitive inhibitors of Gpb, whereas 1 and 3 were inactive. PMID- 9730229 TI - Modifications of the alpha,beta-double bond in chalcones only marginally affect the antiprotozoal activities. AB - Methods for selective alkylation of chalcones in the alpha- or beta-position and for selective reduction of the alpha,beta-double bond have been developed. The antiparasitic potencies of the alpha,beta-double bond modified chalcones only differ marginally from the potencies of the parent chalcones indicating that the propenone residue only functions as a spacer between the two benzene rings, which are the true pharmacophore. PMID- 9730228 TI - Isoxazolo-[3,4-d]-pyridazin-7-(6H)-ones and their corresponding 4,5-disubstituted 3-(2H)-pyridazinone analogues as new substrates for alpha1-adrenoceptor selective antagonists: synthesis, modeling, and binding studies. AB - A series of phenylpiperazinylalkyl moieties were attached to monocyclic or bicyclic substituted pyridazinones and the new compounds tested for their affinity towards alpha1-adrenoceptor and its alpha1a, alpha1b and alpha1d subtypes, as well as serotonin 5-HT1A receptor. Several analogues (5, 6, 9, and 10) showed remarkable potency and selectivity towards alpha1a, and alpha1d with respect to alpha1b subtype. None of the test compounds exhibited significant affinity for 5-HT1A receptor. Finally, on the basis of the alpha1-AR subtypes 3D models recently proposed, we have elaborated theoretical interaction models for the new compounds. The theoretical study allowed us to predict the affinity of the new compounds as well as to infer the structural/dynamics determinants of their interaction with the three alpha1-AR subtypes. PMID- 9730230 TI - Synthesis and binding mode of heterocyclic analogues of suramin inhibiting the human basic fibroblast growth factor. AB - The design, synthesis, and biological evaluation of a series of pyrrole and pyrazole congeners 2 of suramin, directed toward the development and identification of new ligands that complex the human fibroblast growth factor (bFGF), thereby inhibiting tumor-promoted angiogenesis, is reported. Compounds 2 were evaluated for their ability to inhibit binding of bFGF to its receptor, in vivo bFGF-induced angiogenesis, and neovascularization of the chorioallantoic membrane in comparison with suramin. These assays showed that ligands 2 exhibit moderate to good activity, comparable to that of suramin, and are less toxic than suramin itself. In this study, affinity data of ligands in combination with the crystal structure of bFGF were used to explain structure-affinity relationships and to gain an insight into the possible mode of ligand-protein interaction. Due to the lack of experimental structural data on the ligand-bFGF complexes, molecular mechanics techniques were used to obtain putative bioactive conformations and to generate docked complexes with the three-dimensional structure of bFGF. These experiments led to suggest that compounds 2 give rise to 1:1 complexes with bFGF through an unprecedented, bidentate attachment of their naphthylsulfonate groups to two main domains, commonly referred to as the heparin binding site and the receptor binding site, on bFGF, thus preventing the interaction of the growth factor with its receptor. PMID- 9730232 TI - Structural modifications induced during biodegradation of wheat lignin by Lentinula edodes. AB - The structural modifications occurring during wheat straw lignin biodegradation were evaluated by the concerted use of 31P-, 1H- and 2D homo- and heteronuclear NMR spectroscopies. Straw lignin was found to be oxidatively degraded via stereoselective side-chain oxidation as evidenced by a lower erythro/threo ratio. Significantly lower amounts of phenolic hydroxy and methoxy groups in the decayed lignin may be indicative that its structure after the fungal treatment contained a lower amount of aromatic units. In addition an increase in carboxylic acids content, that cannot be explained only on the basis of side-chain oxidation reactions, was also apparent. This evidence, coupled with pertinent data collected during this effort, suggests the occurrence of aromatic ring cleavage reactions. In fact the oxidative degradation of lignin model compounds by fungi has been reported to occur via the 3-oxoadipate pathway which is known to cause aromatic ring cleavage with the formation of aliphatic chains. The presence of aliphatic moieties in the decayed lignin was confirmed by detailed 1H- and 2D NMR spectroscopic analyses. PMID- 9730231 TI - Glabracins A and B, two new acetogenins from Annona glabra. AB - Two new bioactive bis-THF Annonaceous acetogenins, glabracins A (1) and B (2), and two previously known acetogenins, javoricin (3) and bullatanocin (4), have been isolated from the leaves of Annona glabra by activity-directed fractionation using the brine shrimp lethality test (BST). The structures of 1 and 2 were elucidated based on spectroscopic and chemical methods, and the absolute stereochemistries were partially determined by the advanced Mosher ester method. 1 and 2 showed selective cytotoxicities to certain human tumor cell lines, and 1 was significantly more potent although 1 and 2 differ only in the stereochemistry of their vicinal diols at C-23/24. PMID- 9730233 TI - The retro-Mannich cleavage of delta1,delta1'-tryptophan dimers. AB - Under acidic conditions tryptophan sidechains crosslink to form delta1,delta1' tryptophan dimers through a Mannich-type mechanism. Tryptophan dimers are readily cleaved at high temperatures under acidic conditions making it impossible to isolate tryptophan dimers under standard conditions of acidic protein hydrolysis. In a prescriptive sense this cleavage can be used to recover peptides that have undergone tryptophan crosslinking, although the yields drop with increasing peptide length due to competitive cleavage of the amide bonds. The best conditions for cleavage involve heating the dimeric peptides in dilute ethanolic HCl at 150 degrees C in the presence of ten equivalents of ethanedithiol. PMID- 9730234 TI - Synthesis and anticonvulsant properties of triazolo- and imidazopyridazinyl carboxamides and carboxylic acids. AB - Analogues of 3-amino-7-(2,6-dichlorobenzyl)-6-methyltriazolo[4,3-b]pyridazine PC25 containing amide or carboxylic acid function were synthesized and tested for anticonvulsant activity. The compounds having the imidazole ring substituted with an amide group have been found to be generally more active against maximal electroshock-induced seizures in mice (15.2 < or = ED50 < or = 37.5 mg kg(-1) orally). Furthermore, maximum activity was generally associated with a 2,6 dichlorobenzyl substitution pattern. 3-Amido-7-(2,6-dichlorobenzyl)-6 methyltriazolo[4,3-b]pyridazine 4b was also protective in the pentylenetetrazole induced seizures test (ED50 = 91.1 mg kg(-1) orally) and blocked strychnine induced tonic extensor seizures (ED50 = 62.9 mg kg(-1) orally). Moreover, calculated electrostatic isopotential maps of the whole active compounds were quite similar and, consequently, could be associated to optimum anticonvulsant activity. PMID- 9730235 TI - Cross-linking of proteins by Maillard processes--model reactions of D-glucose or methylglyoxal with butylamine and guanidine derivatives. AB - Advanced Maillard reaction in proteins leads to formation of covalently cross linked aggregates the chemical nature of which is largely unknown. From model reactions of methylglyoxal and butylamine with creatine or alpha-N-acetyl-L arginine, one main product each was isolated. These two compounds were identified, on the basis of unequivocal spectroscopic evidence, as 2-[(5 butylimino-4-methyl-4,5-dihydro-1H-2-imidazolyl)(methyl)amin o]acetic acid and 2 acetylamino-5-[(5-butylimino-4-methyl-4,5-dihydro-1H-2-imidazoly l)amino]pentanoic acid, respectively. Using D-glucose instead of methylglyoxal, two main products each were obtained from reaction with the respective guanidine derivative. The spectroscopic data definitively establish the formation of the diastereoisomeric 2-[(4-butyl-6,7-dihydroxy-4,5,6,7,8,8a-hexahydroimidazo[4,5-b]a zepin-2-yl)(methyl)amino]acetic acid from the reaction with creatine, and of the diastereoisomeric 2-acetylamino-5-[(4-butyl-6,7-dihydroxy-4,5,6,7,8,8a hexahydroimidazo [4,5-b]azepin-2-yl)amino]pentanoic acid from the reaction with alpha-N-acetyl-L-arginine. All products were isolated in fairly good yield and represent 1:1:1 adducts of the respective reaction partners. Formation of these compounds thus constitutes an efficient reaction pathway for linking primary amines to guanidine derivatives. It seems justified, therefore, to expect cross linking of proteins by action of reducing carbohydrates to proceed analogously. PMID- 9730236 TI - Binding affinity of Cu(II)-VP-16 (etoposide) complex and its analogues to DNA and hydroxyl radical generation during DNA strand breaks. AB - Conformational effects and affinities of VP-16 (etoposide) and its derivatives to DNA in the presence of Cu(II) ion were examined by circular dichroic (CD) spectra. The Cu(II)/Cu(I) redox kinetics and the hydroxyl radical (.OH) generation from the Cu(II)-complexes were estimated by the stopped-flow kinetics. Based on the results, DNA-cleaving activity of Cu(II)-complexes of VP-16 has been shown to be related with binding affinity of the complex to DNA, Cu(II)/Cu(I) redox and .OH generation, emphasising the mechanism of generated .OH attack to DNA. PMID- 9730237 TI - Novel cephalosporin derivatives possessing a bicyclic heterocycle at the 3 position. Part I: Synthesis and biological activities of 3-(benzothiazol-2 yl)thiocephalosporin derivatives, CP0467 and related compounds. AB - A series of cephalosporin derivatives with various bicyclic heterocycles at the C 3 position was synthesized and evaluated for antibacterial activity. Among them CP0467 (3a) showed excellent antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA) (MIC90 = 6.25 microg/mL), and extremely high affinity for the penicillin binding protein 2' of MRSA (I50 = 0.49 microg/mL). Furthermore, 3a showed a long-acting pharmacokinetic profile in mice (AUC(infinity) = 482.3 microg/h/mL and T(1/2) = 1.9 h). PMID- 9730238 TI - Mode of action of escins Ia and IIa and E,Z-senegin II on glucose absorption in gastrointestinal tract. AB - We examined the mode of action of escins Ia (1) and IIa (2) and E,Z-senegin II (3) for the inhibitory effect on the increase in serum glucose levels in oral glucose-loaded rats. Although 1-3 inhibited the increase in serum glucose levels in oral glucose-loaded rats, these compounds did not lower serum glucose levels in normal or intraperitoneal glucose-loaded rats, or alloxan-induced diabetic mice. Furthermore, 1-3 suppressed gastric emptying in rats, and also inhibited glucose uptake in the rat small intestine in vitro. These results indicated that 1-3 given orally have neither insulin-like activity nor insulin-releasing activity. Compounds 1-3 inhibited glucose absorption by suppressing the transfer of glucose from the stomach to the small intestine and by inhibiting the glucose transport system at the small intestinal brush border. PMID- 9730239 TI - Properties of nicked and circular dumbbell RNA/DNA chimeric oligonucleotides containing antisense phosphodiester oligodeoxynucleotides. AB - We have designed a new class of oligonucleotides, 'dumbbell RNA/DNA chimeric phosphodiester oligonucleotides', consisting of a sense RNA sequence and its complementary antisense DNA sequence, with two hairpin loop structures. The reaction of the Nicked (NDRDON) and Circular (CDRNON) dumbbell DNA/RNA chimeric oligonucleotides with RNase H gave the corresponding antisense phosphodiester oligodeoxynucleotide together with the sense RNA cleavage products. The liberated antisense phosphodiester oligodeoxynucleotide was bound to the target 45 mer RNA, which gave 45 mer RNA cleavage products by treatment with RNase H. The circular dumbbell RNA/DNA chimeric oligonucleotide showed more nuclease resistance than the linear antisense phosphodiester oligodeoxynucleotide (anti-ODN) and the nicked dumbbell RNA/DNA chimeric oligodeoxynucleotide. The circularization, achieved by joining the 3' and the 5' ends of RNA/DNA chimeric oligonucleotides containing two hairpin loop structures, increases the oligonucleotide uptake into cells, as compared with the nicked dumbbell RNA/DNA chimeric oligonucleotide and the linear antisense phosphodiester oligodeoxynucleotides. When the circular dumbbell RNA/DNA chimeric oligonucleotide is directly delivered into retrovirus infected cells, its antisense phosphodiester oligodeoxynucleotide function appears. PMID- 9730241 TI - Syntheses and cytotoxicities of four stereoisomers of muricatacin from D-glucose. AB - Four stereoisomers of muricatacin 1a-d were prepared by the reaction of corresponding aldehydes 4a-d, which in turn were prepared from D-glucose, with the anion of triethylphosphonoacetate followed by reduction and cyclization under acidic conditions. Cytotoxicities of four stereoisomers were tested against in vitro A-549 cell line as well as MCF-7 cell line. Stereochemistry at C4 and C5 position of muricatacin did not affect the cytotoxicities significantly. PMID- 9730240 TI - Pharmacophore identification of a chemokine receptor (CXCR4) antagonist, T22 ([Tyr(5,12),Lys7]-polyphemusin II), which specifically blocks T cell-line-tropic HIV-1 infection. AB - We have previously found that T22 ([Tyr(5,12), Lys7]-polyphemusin II) has strong anti-human immunodeficiency virus (HIV) activity, and that T22 inhibits T cell line-tropic HIV-1 infection mediated by CXCR4/fusin. T22 is an 18-residue peptide amide, which takes an antiparallel beta-sheet structure that is maintained by two disulfide bridges. Structure-activity relationship (SAR) studies on T22 have disclosed the contributions of each region of T22 to activity or cytotoxicity, and have provided the following useful information to develop new CXCR4 antagonists: The number of Arg residues in the N-terminal and C-terminal regions of T22 is closely related to anti-HIV activity. Addition of a variety of functional groups at the N-terminal end results in increases in activity. Disulfide rings, especially the major disulfide loop, are indispensable for anti HIV activity and maintenance of the beta-sheet structure. Trp3 can be replaced by other aromatic residues (Tyr, Phe and L-2-naphthylalanine). Between two repeats of Tyr-Arg-Lys, which are a characteristic structure in T22, Tyr-Arg-Lys in the N terminal portion is more closely associated with anti-HIV activity and maintenance of the beta-sheet structure. A positive charge in the side chain at the (i + 1) position of the beta-turn region is necessary for strong activity. Through these studies, we have found several compounds having higher selectivity indexes (50% cytotoxic concentration/50% effective concentration) than that of T22. PMID- 9730242 TI - Antiallergic activities of rabdosiin and its related compounds: chemical and biochemical evaluations. AB - We examined the effects of caffeic acid-containing compounds such as chlorogenic acid, rosmarinic acid and rabdosiin on anti-allergic activities involving active oxygens scavenging activity as well as inhibitory activities of hyaluronidase and beta-hexosaminidase release. Rabdosiin exhibited the highest hyaluronidase inhibitory activity and scavenging activities against active oxygens species such as superoxide anion radicals and hydroxyl radicals among the tested compounds. Both rabdosiin and caffeic acid inhibited beta-hexosaminidase release from cultured cells more than 90% at 2 mM. The inhibition by rosmarinic acid and chlorogenic acid were weaker than that of rabdosiin. From these results, rabdosiin has been proposed to possess anti-allergic activity. PMID- 9730243 TI - Novel antiallergic agents. Part I: Synthesis and pharmacology of pyrimidine amide derivatives. AB - We have synthesized many pyrimidine amide derivatives. Novel pyrimidine bis glycolic amide derivatives showed moderate inhibition in the rat passive cutaneous anaphylaxis (PCA) assay by oral administration. Among these compounds, 2,4-bis(methoxyacetylamino)-6-piperidinopyrimidine (2i) exhibited significant inhibition. However the compound (2i) did not inhibit antigen-induced histamine or SRS-A release from lung fragments of the guinea-pig at less than 10(-4) M. Derivatives of 2i have also notable or moderate activity in the rat PCA assay. Compound 2h which has no oxygen atom at the alpha-position of the amide carbonyl group and, compound 17 which has no amide carbonyl group, showed no inhibition in the rat PCA assay. We supposed that both the amide carbonyl group and the oxygen atom at alpha-position of the amide carbonyl group play an important role in inhibiting the rat PCA reaction. These pyrimidine bis-glycolic amide derivatives have a novel structure and unique activity which suggests they may be potentially useful in the treatment of allergic diseases. PMID- 9730244 TI - Novel antiallergic and antiinflammatory agents. Part I: Synthesis and pharmacology of glycolic amide derivatives. AB - A series of mono-glycoloylamino derivatives was synthesized by treatment of the corresponding aromatic monoamine derivatives with glycoloyl chloride derivatives in pyridine or dichloromethane, in the presence of a base such as triethylamine or pyridine. Hydrolysis of acetoxy compounds in aqueous ammonia and methanol solution produced hydroxy derivatives with ease. These compounds were tested in the rat PCA (passive cutaneous anaphylaxis) assay by oral administration. Thiazole and thiadiazole derivatives showed moderate inhibition in this assay. In contrast, benzothiazole and benzonitrile derivatives exhibited marked inhibition. In particular, compound 5t also showed marked inhibition of eosinophil adhesion to TNF (tumor necrosis factor) -alpha-treated HUVEC (human umbilical vein endothelial cells) in the range of 10(-8)-10(-5) M. PMID- 9730245 TI - Novel antiallergic and antiinflammatory agents. Part II: Synthesis and pharmacology of TYB-2285 and its related compounds. AB - A series of m-bis(glycoloylamino)benzene derivatives was synthesized by treatment of the corresponding m-diaminobenzene derivatives with glycoloyl chloride derivatives in pyridine. Hydrolysis of acetyl compounds gave hydroxy derivatives, from which other acyl derivatives could be synthesized. These compounds were tested in the rat PCA (passive cutaneous anaphylaxis) assay by oral administration. Benzonitrile derivatives (4c, 5c, 6c, 4h, 5h) exhibited notable inhibition in this assay. Compounds 5c and 6c also showed remarkable inhibition of eosinophil adhesion to TNF- (tumor necrosis factor) alpha-treated HUVEC (human umbilical vein endothelial cells) in the range of 10(-8)-10(-5) M. Compound 5c is now under investigation in Japan as TYB-2285 (Figure 1) for asthma and atopic dermatitis in phase II clinical studies. PMID- 9730246 TI - Novel dithiocarbamate carbapenems with anti-MRSA activity. AB - A new series of 1beta-methyl carbapenems, in which a disubstituted aminothiocarbonylthio moiety was attached to the C-2 position of the carbapenem nucleus, were prepared and evaluated for anti-MRSA activity. These derivatives showed good in vitro antibacterial activity against high-level MRSA, and the finding of good affinity for PBP-2' supported these results. Some of the compounds having favorable protein-binding affinity showed excellent in vivo anti MRSA activity. PMID- 9730247 TI - Structure-cytotoxic activity relationship for the toad poison bufadienolides. AB - The toad poison bufadienolides including natural and derivatized compounds were tested for their cytotoxic effects on primary liver carcinoma cells PLC/PRF/5 and their structure-cytotoxic activity relationships were studied. For this study, a ligand-binding model was developed by using a pharmacophore mapping program, Distance Comparisons (DISCO). The structural features that are common to the 3D structures of active bufadienolides were identified to provide approach to a 3D QSAR method by using Comparative Molecular Field Analysis (CoMFA) study and to correlate the steric and electrostatic fields of the molecules to their activities. A valuable model which enables prediction of their activities was obtained from the CoMFA analysis, which may be employed for the drug designs of new bufadienolide analogues. PMID- 9730248 TI - Photoaffinity labeling of PKC isozymes by phorbol ester derivatives. AB - Photoaffinity probes PPDA and PPTD, which have diazoacetyl and trifluorodiazopropionyl group at C-13 position in phorbol, respectively, were synthesized. Photoaffinity labeling of protein kinase C isozymes by both the probes resulted in specific cross-linking. PMID- 9730249 TI - Immunopharmacologic agents in the amelioration of hepatic injuries. AB - A number of immunomodulating agents of different origin have been shown to reduce liver injury of various etiologies. Immunostimulants like levamisole, BCG, a protein polysaccharide from myceria Coriolus vesicolor PS-K, a streptoccocal preparation OK-432 and immunomodulators like N-acetylmuramyl-L-alanyl-D isoglutamine (MDP) and its analogs. Selective T-cell suppressors like the polypeptide cyclosporine A (CsA) and the macrolide FK 506 (tacrolimus) have also been claimed to possess hepatoprotrophic or hepatoprotective properties at low doses. The aim of this review article is to highlight the interplay between the administration of immunomodulating agents and the amelioration of hepatic injuries. Hepatic effects of exogenous immunomodulators are discussed with special focus on the most widely used immunosuppressive agents, CsA and tacrolimus. An important question exists as to whether these potential hepatoprotective effects are related mechanistically to the immune system or are working at different levels. Due to the differences in effects and modes of actions of various immunoactive substances presented herein, a common mechanism for their cytoprotective effects cannot be formulated at this stage. Levamisole and cyanidanol may protect cells against necrosis by acting as free radical scavengers. MDP and its analogs reduce carbon tetrachloride-elevated (CCl4) lipid peroxides and their protective effects are primarily on hepatic cytoplasmic membranes where lipid peroxidation and calcium homeostasis interact. MDP reduced CCl4-elevated calcium in both intact hepatocytes and in the post microsomal supernatant suggest that the influx of extracellular calcium across plasma membrane is affected. Elevations of intracellular calcium above a threshold are involved in: the stimulation of Ca2+-sensitive enzymes such as phospholipase A2, endonucleases and proteases, the conversion of xanthine dehydrogenase to xanthine oxidase and the formation of free radicals, all of which disturb biomembranes. MDP and its analogs, in a specific dose range, may act to maintain intracellular calcium within physiological ranges. Highly complex cellular signalling systems, including calcium, are involved in the explanation of the mechanism of the immunosuppressive effect of CsA and tacrolimus. The hepatoprotective effects of these selective immunosuppressive agents, however, are independent of the inhibition of T-cell activation. The cyclophilin and tacrolimus binding proteins of the mitochondria are the receptors for these compounds and play a key role in the regulation of mitochondrial permeability transition pores. CsA or tacrolimus inhibition of mitochondrial permeability transition pores does not require interaction with calcineurin, indicating a dissociation between immunosuppression and mitochondrial protection. The involvement of intracellular or intramitochondrial proteins in the modulation of mitochondrial permeability transition pores with the creation of a partially impermeable state for Ca2+ movement in drug-treated mitochondria and the dissociation of this effect from immunomodulatory actions potentially offers new and promising approaches for the development of new pharmacologicals targeted at therapeutic intervention. Clinical trials of these drugs as hepatoprotective agents are limited. Use of CsA in patients with primary biliary cirrhosis and autoimmune chronic hepatitis and in cirrhotic animal models produced by chronic administration of CCl4 have yielded encouraging results. It seems that this class of compounds may be of substantial benefit in liver protection against many pathological conditions where disturbance in mitochondrial function and in Ca2+ homeostasis appear to be prerequisites for cell injury. PMID- 9730250 TI - Suppression of in vitro IFN-gamma production by spleen cells of Plasmodium chabaudi-infected C57BL/10 mice exposed to dexamethasone at a low dose. AB - After infection with Plasmodium chabaudi, C57BL/10 mice produced significant amounts of serum IFN-gamma, developed a low level of parasitemia and survived the infection. Production of IFN-gamma was also obvious when spleen cells of the infected mice were stimulated with the parasite antigen contained in an erythrocyte lysate in vitro. Depletion of CD4+ T cells abrogated production of IFN-gamma, leading to loss of resistance to the infection. The CD4+ T cells/IFN gamma-dependent resistance of the C57BL/10 mice against P. chabaudi was applied to assess immunotoxicological effect of dexamethasone (DEX). Administration of a high dose (0.75 mg/kg) resulted in loss of the splenic cellularity, remarkable decrease in serum IFN-gamma production, increased level of parasitemia, and eventual death of the infected mice. In contrast, DEX at a low dose (0.02 mg/kg) induced no alternation in the in vivo host immune activity and the mice survived the infection. However, when spleen cells were obtained from the infected mice administered the low dose of DEX and stimulated in vitro with the parasite antigen, a significantly decreased level of IFN-gamma was demonstrated with compared to control mice. These findings demonstrate that the in vitro production of IFN-gamma by spleen cells from P. chabaudi-resistant C57BL/10 mice was more sensitive to the immunosuppressive effect of in vivo administration of DEX. This ex vivo assay might provide a method for evaluation of drug-induced immunotoxicity at a higher sensitivity than the conventional host resistant assays such as comparison of severity of disease or time to death. PMID- 9730251 TI - Inhibitory effect of Asparagus cochinchinensis on tumor necrosis factor-alpha secretion from astrocytes. AB - We investigated whether an aqueous extract of Asparagus cochinchinensis Merrill (Liliaceae) roots (ACAE) inhibits secretion of tumor necrosis factor-alpha (TNF alpha) from primary cultures of mouse astrocytes. ACAE dose-dependently inhibited the TNF-alpha secretion by astrocytes stimulated with substance P (SP) and lipopolysaccharide (LPS). IL-1 has been shown to elevate TNF-alpha secretion from LPS-stimulated astrocytes while having no effect on astrocytes in the absence of LPS. We therefore investigated whether IL-1 mediated inhibition of TNF-alpha secretion from astrocytes by ACAE. Treatment of ACAE to astrocytes stimulated with both LPS and SP decreased IL-1 secretion. Moreover, incubation of astrocytes with IL-1 antibody abolished the synergistic cooperative effect of LPS and SP. These results suggest that ACAE may inhibit TNF-alpha secretion by inhibiting IL 1 secretion and that ACAE has a antiinflammatory activity in the central nervous system. PMID- 9730252 TI - Prophylactic effect of Korean mistletoe (Viscum album coloratum) extract on tumor metastasis is mediated by enhancement of NK cell activity. AB - We here demonstrated the prophylactic effect of an extract (KM-110) from Viscum album coloratum, a Korean mistletoe, on tumor metastasis produced by highly metastatic tumor cells, colon 26-M3.1 carcinoma, B16-BL6 melanoma and L5178Y-ML25 lymphoma cells, using experimental models in mice. Intravenous (i.v.) administration of KM-110 (100 microg/mouse) 2 days before tumor inoculation significantly inhibited lung metastasis of B16-BL6 and colon 26-M3.1 cells, and liver and spleen metastasis of L5178Y-ML25 cells. The prophylactic effect of KM 110 on tumor metastasis was evident with various administration routes, i.e. subcutaneous, oral, intranasal as well as i.v., and was dependent upon the dose of KM-110 administered. Furthermore, mice given KM-110 (100 microg) 2 days before tumor inoculation showed significantly prolonged survival rates compared with the untreated mice. In a time course analysis of NK activity, i.v. administration of KM-110 (100 microg) significantly augmented NK cytotoxicity to Yac-a tumor cells from 1 to 3 days after KM-110 treatment. Furthermore, depletion NK cells by injection of rabbit anti-asialo GM1 serum completely abolished the inhibitory effect of KM-110 on lung metastasis of colon 26-M3.1 cells. These results suggest that KM-110 possesses immunopotentiating activity which enhances the host defense system against tumors, and that its prophylactic effect on tumor metastasis is mediated by NK cell activation. PMID- 9730253 TI - The effect of theophylline on blood and sputum eosinophils and ECP in patients with bronchial asthma. AB - It was recently reported that theophylline has an anti-inflammatory and bronchodilating effect on bronchial asthma. Accordingly, to examine the anti inflammatory effect of theophylline on asthma, especially its effect on eosinophil activation, a sustained-release theophylline preparation (Theolong) was administered (daily dose: 400 mg) to 18 patients with mild to moderate bronchial asthma. This was done in order to study the preparation's effects on lung function, blood and sputum eosinophils and ECP four weeks pre- and post administration. Lung function was determined by spirometry and sputum by induced sputum. Blood and sputum ECP levels were determined using an ECP RIA kit. In lung function, there were no differences in vital capacity (VC) or in forced expiratory volume 1 s (FEV 1.0) pre- and post-administration. There were also no differences in the number of blood and sputum eosinophils, but serum and sputum ECP levels decreased. Theophylline is thus expected to exert an inhibitory effect on eosinophil activation and it is suggested as an effective therapeutic drug for bronchial asthma. PMID- 9730254 TI - Effect of combined Cyclosporine A and liposome encapsulated dichloromethylene diphosphonate on the organisation of the rat thymus: evidence for a role of macrophages in guiding the post Cyclosporine A thymic reorganisation. AB - Cyclosporine A (CsA) is a powerful immunosuppressant inducing marked involution of the thymic medulla, and disappearance of interdigitating cells (IDCs) and reducing the number of macrophages (Mphi). Usually, while the thymus of rats receiving a short course of CsA promptly recovers after stopping CsA treatment, long term CsA treatment, like mediastinal irradiation, impairs the normal thymic recovery and is thought to be responsible for the development of autoimmune diseases. In the present study we evaluated the role played by the IDCs and Mphi in the normal recovery of the thymic histology at light and ultrastructural level. Besides CsA administration, we also used liposome-encapsulated dichloro methylene-diphosphonate (lip-CL2MDP), that induces a total depletion of the Mphi resistant to CsA. After a short (21 days) course of CsA and lip-CL2MDP administration, we did not observe the normal recovery of the thymic parenchyma but only cortical zones consisting of lymphoblasts, epithelial cells and Mphi. The CsA/lip-CL2MDP treatment determining the loss of IDCs and Mphi and consequently the loss of the normal thymic histology seems to simulate in the rats, the long term CsA treatment or the mediastinal irradiation. The results obtained suggest that the loss of IDCs and the depletion of Mphi interfere with the normal thymic recovery. The delay in the recovery of I DCs could be a consequence of the absence of macrophages. These findings would indicate that the IDCs, determining the negative selection of T-lymphocytes, are the main cells responsible for the thymic microenvironment. PMID- 9730255 TI - Enhancement of phagocytosis in mouse peritoneal macrophages by fragments of acidic fibroblast growth factor (aFGF). AB - To characterize the effects of acidic fibroblast growth factor (aFGF) in mouse peritoneal macrophages, the effects of aFGF fragments on phagocytosis were examined. Fragments that were tested included aFGF(1-15), aFGF(1-20), aFGF(1-29), Ala16-aFGF(1-29), aFGF(9-29) and aFGF(114-140). aFGF(1-29) induced an enhancement of phagocytosis in a dose-dependent manner and was more effective than any other fragments tested. Even in Ca2+-and Mg2+-free solutions, phagocytosis was enhanced by aFGF(1-29). However, the enhancement induced by aFGF(1-29) was completely inhibited in the presence of mannan (4 mg/ml). Furthermore, the enhancement of phagocytosis by aFGF(1-29) was suppressed by heparin (100 microg/ml). The results of the present study suggest that the active region of aFGF that is responsible for the enhancement of phagocytosis corresponds to residues 15-29 and that phagocytosis, which is modulated by aFGF, is independent of extracellular Ca2+ and is mediated by mannose receptors. PMID- 9730256 TI - Estrogenic xenobiotics affect the intracellular activation signal in mitogen induced human peripheral blood lymphocytes: immunotoxicological impact. AB - The present study was an attempt to elucidate the effect of estrogenic xenobiotics on the proliferation of mitogen-stimulated human peripheral blood lymphocyte (PBL). Our findings follow: (a) the proliferation of PBL in response to phytohemagglutinin (PHA) was mediated by protein kinase C activity, but estrogenic xenobiotics had a strong inhibitory effect on protein kinase C activity of PHA-stimulated PBL; (b) cytoplasmic extracts from PHA-stimulated PBL greatly activated DNA replication, but estrogenic xenobiotics had a strong inhibitory effect on these activities. The results suggest that the cytoplasmic signal-generating system in mitogen-treated PBL is inhibited by estrogenic xenobiotics, and that the defect occurs at all stages in the sequence of events leading to DNA synthesis and cell proliferation. PMID- 9730257 TI - Effects of immunopharmacological compounds in the Lyme arthritis model using normal and SCID mice. AB - The murine Lyme borreliosis causes a special type of arthritis whose development appears to be controlled by a functioning immune system. Immunocompetent C3H and severe combined immunodeficient (SCID) mice were infected with Borrelia burgdorferi (strain SH-2-82) to induce experimental Lyme disease. Expression of clinical symptoms was mild to very moderate in the C3H but more rapidly developing and severe in the SCID mouse. Various pharmacological compounds, such as anti-inflammatory and immunosuppressive drugs, monoclonal antibodies and other miscellaneous agents, were investigated for profiling their effects in this model in both mouse strains. Several disease parameters were assessed, in particular paw swelling. The use of these various compounds provided further evidence that experimental borreliosis in mice represents a special type of arthritis which has no autoimmune basis and which requires productive infection with Borrelia burgdorferi. In addition, when comparing these results with those obtained in other mainly immune driven arthritis models commonly used in inflammation research, it is concluded that this arthritis model is not suitable for the therapeutic assessment of antiinflammatory agents. PMID- 9730258 TI - Effect of Rehmannia glutinosa on immediate type allergic reaction. AB - We investigated the effect of aqueous extract of Rehmannia glutinosa steamed root (RGAE) on the allergic reactions in vivo and in vitro. RGAE dose-dependently inhibited systemic allergic reaction induced by compound 48/80. When RGAE was pre treated at the same concentrations with systemic allergic reaction test, the plasma histamine levels were reduced in a dose-dependent manner. RGAE dose dependently inhibited skin allergic reaction activated by anti-dinitrophenyl (DNP) IgE. RGAE also dose-dependently inhibited the histamine release from the rat peritoneal mast cells (RPMC) by compound 48/80 or anti-DNP IgE. Moreover, RGAE had significant inhibitory effect on anti-DNP IgE-induced tumor necrosis factor-alpha production of RPMC. These results indicate that RGAE may be beneficial in the regulation of immediate type allergic reaction. PMID- 9730259 TI - Inhibition of anti-CD3 antibody-induced mouse T cell activation by pentoxifylline in combination with rapamycin or A77 1726 (leflunomide). AB - Pentoxifylline (PTX), rapamycin (RAP), and leflunomide are potent immunomodulatory drugs with differing modes of action. In order to develop new drug combinations for immunotherapy, we tested the effects of PTX in combination with RAP or A77 1726 (the active metabolite of leflunomide) on in vitro T cell activation in a mouse model system. T lymphocytes in spleen cell preparations were stimulated with anti-CD3 monoclonal antibody alone, or in the presence of PTX (25-200 microg/ml), RAP (0.5-5.0 ng/ml), A77 1726 (2.5-10.0 microM), PTX/RAP (25-200 microg/ml and 0.5-5.0 ng/ml, respectively), or PTX/A77 1726 (25-200 microg/ml and 2.5-10.0 microM, respectively). Anti-CD3-induced T cell proliferation was inhibited in a dose-dependent fashion by the individual drugs. An additive inhibitory effect was observed in cultures treated with PTX/RAP or PTX/A77 1726. The effects of PTX, RAP, A77 1726, PTX/RAP, or PTX/A77 1726 (at concentrations approximating the IC50 of individual drugs for inhibition of lymphoproliferation) on anti-CD3-activated killer (AK) cell induction, CD25 expression, and interleukin (IL)-2 synthesis in anti-CD3-activated spleen cell cultures were also determined. Alone, each drug was able to suppress AK cell induction to varying degrees. PTX plus RAP exhibited strong synergism, while the combination of PTX and A77 1726 had an additive inhibitory effect on AK cell induction. CD25 expression was only weakly inhibited by A77 1726, but the percentage of CD25-expressing cells was greatly reduced in cultures treated with PTX or RAP. The combination of PTX and RAP had an additive inhibitory effect on CD25 expression while PTX and A77 1726 together had an effect equivalent to PTX alone. IL-2 synthesis was inhibited by PTX but was unaffected by RAP or A77 1726. Treatment with PTX plus RAP led to a further reduction in IL-2 production but co treatment with PTX and A77 1726 approximated the inhibitory effect of PTX alone. We conclude that the combination of PTX and RAP is noteworthy for its potent immunomodulatory activity and may be of use in clinical situations where it is desirable to prevent T cell activation. PMID- 9730260 TI - Involvement of the L-arginine/nitric oxide neural pathway in non-adrenergic, non cholinergic relaxation of the bovine oesophageal groove. AB - 1. The distribution and localization of nitric oxide synthase (NOS) immunoreactivity and nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) activity in the bovine oesophageal groove were investigated by immnunohistochemical and histochemical staining techniques. Functional in vitro studies were performed to correlate the presence of NOS-immunoreactivity (IR) and NADPH-d staining with smooth muscle relaxations involving the L-arginine/nitric oxide neural pathway in the bovine oesophageal groove activity. 2. NOS-IR and NADPH-d were expressed in nerve cell bodies of the myenteric, submucosal and intramuscular ganglia, and in nerve fibres distributed around blood vessels and throughout the different muscular layers of the bovine oesophageal groove. 3. In oesophageal groove strips treated with guanethidine (10(-5) M) and atropine (10( 7) M) to block noradrenergic neurotransmission and muscarinic receptors, respectively, electrical field stimulation (EFS, 0.5-32 Hz, 1 ms duration, 20-s trains) induced relaxations which were practically abolished by tetrodotoxin (TTX, 10(-6) M). 4. Incubation with an inhibitor of nitric oxide synthesis, NG nitro-L-arginine (L-NOARG, 3 x 10(-5) M), significantly inhibited relaxations induced by EFS. This inhibition was partially reversed by L-arginine (L-arg, 5 x 10(-3) M). D-NOARG (3 x 10(-5) M) had no effect on EFS-induced relaxations. 5. NO added as an acidified solution of NaNO2 (10(-6) - 10(-3) M) and S-nitroso-L cysteine (10(-7) - 10(-4) M) caused concentration-dependent relaxations of the bovine oesophageal groove. These relaxations were unaffected by L-NOARG (3 x 10( 5) M). 6. The presence of NO-synthesizing enzyme in nerves and ganglia, and the pharmacological evidence for NO-mediated smooth muscle relaxation suggested that the L-arg/NO neuronal pathway is involved in the inhibitory neurotransmission of the bovine oesophageal groove. PMID- 9730261 TI - Functional evaluation of 5-hydroxytryptamine receptor activity in rat resistance vessels. AB - 1. To characterize 5-hydroxytryptamine (5-HT) receptors in rat perfused mesenteric vascular bed (MVB), the effect of 5-HT and related compounds was investigated by functional assay. 2. In quiescent preparations, 5-HT elicited a concentration-dependent conctractile response. After addition of ketanserin, a 5 HT2 receptor antagonist, EC50 values were significantly higher than in controls. 3. In noradrenaline (NA)-precontracted preparations, under continuous infusion of ketanserin, 5-HT, 5-carboxamidotryptamine (5-CT) and sumatriptan produced relaxation. Their rank order of relaxant potency and maximum effect were sumatriptan > 5-HT > 5-CT. Methysergide (1 microM) and spiperone (20-100 nM) caused a rightward shift of the relaxation curve to sumatriptan. These data suggest that vasodilatation in rat MVB is mediated by an 'atypical' subtype of 5 HT1-like receptor, which reveals a pharmacological profile similar to that of the 5-HT1D receptor. The involvement of both 5-HT3 and 5-HT4 receptors can be ruled out, since tropisetron (up to 10 microM) was not able to antagonize the relaxant effect by sumatriptan. 4. Under granisetron infusion (3 microM), the contractile response evoked by perivascular nervous stimulation, but not exogenous NA contraction, was significantly reduced (P < 0.001). These data demonstrate the presence of 5-HT3 receptors in peripheral neurones, modulating neurotransmitters release. PMID- 9730262 TI - Cardiac anticholinergic effects of procainamide and its N-acetylated metabolite: experimental pharmacological and radioligand binding studies. AB - 1. The cardiac anticholinergic effects of procainamide (1 mg kg(-1) min(-1)) and its N-acetylated metabolite (NAPA) at equimolar dose (1.16 mg kg(-1) min(-1)) were studied using in vivo experimental pharmacological and in vitro radioligand binding studies. 2. Procainamide and NAPA progressively reduced vagal stimulation induced bradycardia in chloralose-anaesthetized dogs. As indicated by the ED50, the vagolytic activity of NAPA is 1.5-2.0 times weaker than that of procainamide. Both drugs increased heart rate, with lowering of mean blood pressure during the second part of procainamide infusion, but not during NAPA infusion. 3. Binding studies on rat heart membranes yielded Ki values that were 1.5 times higher for NAPA than for procainamide. 4. These results show that NAPA exerts a weaker cardiac vagolytic action than procainamide, which is probably linked to a lower ability to bind to cardiac muscarinic receptors. PMID- 9730263 TI - Dopamine D2-like receptors in the kidney of spontaneously hypertensive rats: a radioligand binding assay and light microscope autoradiography study. AB - 1. Dopamine D2-like receptors were investigated in sections of kidney from male spontaneously hypertensive rats (SHRs) at 6 and 14 weeks of age using radioligand binding assay and autoradiographic techniques with [3H]-spiperone as a ligand. 2. Systolic blood pressure values were slightly higher in 6-week-old SHRs in comparison with age-matched normotensive Wistar-Kyoto (WKY) rats and considerably higher in 14-week-old SHRs in comparison with the other groups investigated. Renal dopamine levels were higher in SHRs aged 6 and 14 weeks in comparison with age-matched WKY rats. Noradrenaline concentrations were similar in 6-week-old SHRs and normotensive WKY rats, and increased slightly in SHRs aged 14 weeks. 3. The density of [3H]-spiperone binding sites was similar in SHRs and WKY rats at 6 weeks of age, and decreased in SHRs at 14 weeks in comparison with age-matched normotensive animals. Light microscope autoradiography revealed the accumulation of silver grains in the tunica adventitia, in the adventitia-media border of intrarenal arteries and within cortical tubules. A few specific silver grains were also developed in the glomerular tuft. No changes in the density and pattern of silver grains were noticeable between SHRs and WKY rats at 6 weeks of age, whereas a reduction in silver grains largely affecting vascular binding sites was observed at 14 weeks of age. 4. Renal denervation considerably decreased the density of [3H]-spiperone binding sites in sections of rat kidney, with an almost complete loss of vascular binding sites. 5. The above findings indicate the occurrence of a decrease of dopamine D2-like receptors in the kidney of SHRs with the progress of hypertension. Dopamine D2-like receptors which are mainly prejunctional are involved in the modulation of sympathetic neurotransmission in the kidney. The loss of these receptors in SHRs may contribute to the pathophysiology of hypertension. PMID- 9730264 TI - Effect of exogenous adenosine and its monophosphate on the contractile response to acetylcholine in the human isolated detrusor muscle strips. AB - 1. Adenosine (0.1-1 mM) or its 5'-monophosphate (5'-AMP) induced a concentration dependent relaxation of tension caused by acetylcholine (0.2 microM) in human urinary bladder detrusor strips. 2. This effect was antagonized concentration dependently by theophylline at an apparent pA2 value of about 5. 3. Maximum relaxation by adenosine or 5'-AMP never exceeded 50% and 80%, respectively, of acetylcholine-induced tension. Relaxation by some beta2-adrenoceptor agonists (0.1-0.2 mM) or norepinephrine was limited to about 50% of maximum. 4. The responses to adenosine and terbutaline were additive, causing full relaxation, and suggesting mobilization of distinct mechanisms underlying muscle relaxation. PMID- 9730265 TI - The L-arginine inhibition of rat middle cerebral artery contractile responses is mediated by inducible nitric oxide synthase. AB - 1. The effect of L-arginine (L-Arg), the nitric oxide synthase (NOS) substrate, on the responses to prostaglandin F2alpha (PGF2alpha, 10 microM) and K+ (120 mM) in rat middle cerebral artery (MCA) segments was analysed. 2. PGF2alpha induced a stable contraction of 0.35+/-0.06 mN mm(-1); the subsequent addition of bradykinin (BK, 1 microM) produced a relaxation of 42+/-9% of the PGF2alpha induced tone. K+ induced a response consisting of a rapid basal tone increase (1.42+/-0.16 mN mm(-1)) followed by a decrease to a stable phase (1.24+/-0.15 mN mm(-1)). 3. L-Arg (0.1 mM), but not D-Arg, decreased the basal tone and reduced the contraction to PGF2alpha in segments with and without endothelium. The contractile response to K+ was also reduced and not maintained in the presence of L-Arg. 4. The inhibitory effect of L-Arg on the PGF2alpha- and K+-induced contractions was completely reversed by the NOS inhibitor, NG-monomethyl-L arginine (L-NMMA, 0.1 mM). 5. Pre-incubation of segments with dexamethasone (1 microM), to inhibit inducible NOS (iNOS), or with the antibiotic polymyxin B (10 microg ml(-1)) reduced the L-Arg inhibition, whereas it was increased by lipopolysaccharide (LPS, 100 ng ml(-1)), an inductor of iNOS. L-NMMA antagonized the effects of dexamethasone and LPS. 6. The present results suggest that L-Arg inhibition of the PGF2alpha- and K+-induced contractions in rat MCA is the result of NO synthesis by iNOS stimulation. PMID- 9730266 TI - Pharmacological characterization and autoradiographic localization of a putative dopamine D4 receptor in the heart. AB - 1. The pharmacological profile and the anatomical localization of a putative dopamine D4 receptor were assessed in sections of rat and human atria and ventricles using combined radioligand binding and autoradiographic techniques with [3H]-spiperone as a ligand. 2. [3H]-Spiperone was bound specifically to sections of human and rat atria and ventricles. The binding was time-, temperature- and concentration-dependent, belonging to a single class of high affinity sites. In atria, the dissociation constant value (Kd) was 0.45 nM in rats and 0.32 nM in humans, and the maximum density of binding sites (Bmax) was 31.6+/-2.9 fmol mg(-1) tissue in rats and 18.8+/-0.7 fmol mg(-1) tissue in humans. In ventricles, Kd was 0.38 nM in rats and 0.39 nM in humans, and the Bmax was 43.5+/-3.0 fmol mg(-1) tissue in rats and 56.4+/-3.2 fmol mg(-1) tissue in humans. 3. The pharmacological profile of [3H]-spiperone binding to sections of both rat and human atria and ventricles was consistent with the labelling of dopamine D2-like receptors. [3H]-Spiperone binding was more sensitive to displacement by the neuroleptic clozapine in sections of atria than of ventricles, suggesting the expression of a dopamine D4 receptor in atrial tissue. Moreover, preincubation of some sections with a dopamine D4 receptor antibody and subsequent exposure to [3H]-spiperone caused a remarkable decrease of radioligand binding to sections of atria, but only a slight reduction of binding to sections of ventricles. 4. Light microscope autoradiography revealed the accumulation of silver grains over atrial tissue within atrial myocardiocytes. A higher density of silver grains was developed in rat than in human atria. In ventricles, silver grains were accumulated primarily in cluster areas both in rats and in humans. 5. The above findings suggest the expression of a dopamine D4 receptor in rat atria, but not in ventricles. A similar site was identified in human atria. The possible relevance of a dopamine D4 receptor in the heart is discussed. PMID- 9730267 TI - The role of MAO-A and MAO-B in the metabolic degradation of noradrenaline in human arteries. AB - 1. Segments of human cystic, gastric and ileocolic arteries were obtained from patients undergoing surgery. 2. Segments of arterial tissues, the noradrenaline content of which ranged between 0.27 and 0.52 microg g(-1), were incubated with 0.1 micromol l(-1) [3H]-noradrenaline for 30 min and the accumulation of the amine as well as the formation of metabolites was measured. 3. In all the arteries, oxidative deamination predominated over O-methylation; the mean values of the deaminated and O-methylated metabolites formed for the three arteries were 247.6 and 82.4 pmol g(-1) tissue, respectively. Dihydroxymandelic acid (DOMA) was the most abundant metabolite. 4. Both clorgyline (a selective MAO-A inhibitor) and selegiline (a selective MAO-B inhibitor) reduced the formation of dihydroxyphenylglycol (DOPEG), DOMA and O-methylated-deaminated metabolites (OMDA), and increased that of normetanephrine (NMN). However, clorgyline depressed the formation of DOPEG more than that of DOMA, while selegiline depressed the formation of DOMA more than that of DOPEG. 5. In conclusion, three major differences distinguish the metabolism of noradrenaline by human arteries from that observed in other species: (1) the large predominance of deamination over O-methylation; (2) the extremely high formation of DOMA; and (3) the relative lack of selectivity of clorgyline and selegiline for MAO-A and B, respectively. Since the arterial vessels used were collected from patients older than 60 years, the morphological changes depending on age may explain the increase in DOMA formation. PMID- 9730268 TI - The influence of compressive loading on growth of cartilage of the mandibular condyle in vitro. AB - The purpose of this study was to clarify the change in mandibular condyles under compressive loading. An organ-culture system of fetal rat mandibular condyles was used, and mechanical loading was generated by compressing the gas phase within a closed chamber. After the culture period, with compressive loading, type I collagen and fibronectin were observed in the lower half of the hypertrophic chondrocyte layer in the mandibular condyles; in contrast, without compressive loading, there was no such reaction. The size of the condyle was not increased by compressive loading. These results suggest that intermittent compressive loading could induce type I collagen and fibronectin production by chondrocytes. PMID- 9730269 TI - Differences in fine control of forces generated by the tongue, lips and fingers in humans. AB - This study compared the fine control of forces generated by the tongue, lips and fingers in middle-aged adults. The aims were to determine whether (1) the articulatory organs (tongue, lips) and fingers differed in the manner of motor control, (2) force control of the various articulatory organs was similar, and (3) control of forces generated by males was different from that of forces generated by females. The relation among several variables of the ramp-and-hold force contraction and target force level was quantified for the articulatory organs and the fingers in 14 normal individuals (7 males and 7 females). Using visual feedback, participants produced ramp-and-hold compression forces as rapidly and accurately as possible to targets ranging from 0.25 to 2 N. The results showed differences in the profiles of forces generated by the articulatory organs and fingers. In particular, the forefingers were characterized by a greater accuracy of force control and precision of movement, a greater stability of the hold phase, but by slower velocities than the articulatory organs. Motor control of the lower lip differed from that of the upper lip and tongue. Mostly, the lower lip was characterized by a greater precision of contraction, faster development of the force, and greater stability of the hold phase than the upper lip and tongue. Gender was a distinguishing factor in the force task; males were able to produce forces with higher velocities and greater precision than females. PMID- 9730270 TI - Morphometric analysis of developing crowns of maxillary primary second molars and permanent first molars in humans. AB - The purpose of this study was firstly to characterize the changes occurring in size and form of the mineralizing maxillary second primary molar and first permanent molar crowns, and secondly to determine if similar changes in size and form characterize enamel apposition in the crowns of these teeth. Twenty-five primary second molars and 20 maxillary permanent first molars at various stages of development, found in archaeological excavations in Israel, were examined for a number of measured variables using image analyser software. Teeth were divided into two groups according to their stage of development: stage I included all teeth at an early stage of development in which mesiobuccal-cusp height was less than 5 mm for the primary molar and 5.9 mm for the permanent molar; stage 2 included all teeth in later stages of development where mesiobuccal-cusp height was greater than these values. In the primary molar, a significant increase was found between the two stages in almost all variables. Significant correlations were also found between all intercusp distances and the external variables. Strong correlations between height of the mesiobuccal cusp and all external and internal variables were noted in stage 1, but fewer in stage 2. In the permanent tooth, no increase was observed in intercusp distances and very few correlations were found between and among the variables. The results suggest that a change in the shape of the maxillary primary second molar occurs during formation, with the lingual cusp tips moving lingually and distally, and the distobuccal cusp tips moving distally. No change occurs in the shape of the maxillary permanent first molar during crown formation. Growth of the maxillary primary second and permanent first molar crowns occurs in 'bursts' of development. PMID- 9730271 TI - Procrustes, Euclidean and cephalometric analyses of the morphology of the mandible in human Class III malocclusions. AB - The role of mandibular phenotype in the development of Class III malocclusion remains unclear. The purpose of this study was to determine whether the form of the mandible differed between prepubertal individuals with Class I and Class III malocclusions. Lateral cephalographs of 73 children of European-American descent aged between 5-11 years with Class III malocclusion were compared to those of 60 counterparts with a normal, Class I molar occlusion. The cephalographs were traced and checked, and eight homologous mandibular landmarks were digitized. Average mandibular geometries, scaled to an equivalent size, were generated using Procrustes superimposition. Euclidean distance matrix analysis (EDMA) was undertaken to corroborate the Procrustes analysis, and bivariate analysis utilizing eight linear and five angular measurements was also performed. Residuals and F-values from Procrustes analysis indicated that mandibular configurations differed statistically for Class I and Class III types. EDMA confirmed that the Class I and Class III geometries were significantly different, revealing that the greatest differences in morphology arose in the anterior-most mandibular regions. As well, most variables showed statistically significant differences when the Class I and Class III mandibular types were compared. When the sample was subdivided into seven age- and sex-matched groups, nearly all age based comparisons were significantly different. It is concluded that the morphology of the mandible differs in individuals with Class III malocclusions when compared to the normal Class I configuration, and that these alterations may indicate dichotomous postnatal mandibular ontogeny. PMID- 9730272 TI - Immunohistochemical findings type I and type II collagen in prenatal mouse mandibular condylar cartilage compared with the tibial anlage. AB - In growing animals the mandibular condylar cartilage serves not only as an articular but also as a growth cartilage, yet, condylar cartilage has some characteristic features that are not found in growth cartilage. For example, some reports suggest that type I collagen, which is not seen in the growth plate cartilage of long bones, is present in the extracellular matrix of condylar cartilage postnatally. Here, the condylar and limb bud cartilage of fetal mice was examined. The distribution of type I and type II collagen in condylar cartilage was already different from that in the limb bud at the first appearance of the cartilage. Type I collagen was demonstrated in the extracellular matrix of the condylar cartilage that first appeared on day 15 of gestation. However, the reaction for type II collagen was much weaker than that for type I collagen. On day 18 of gestation, type I collagen was still found throughout the cell layers but became gradually weaker with depth. Type II collagen was limited exclusively to the deeper layers at this stage. These findings are different from those in the limb bud cartilage, indicating a characteristic feature of the cells in the condylar cartilage present from the prenatal period. PMID- 9730273 TI - Effect of testosterone replacement after neonatal castration on craniofacial growth in rats. AB - Neonatal castration precludes the pubertal increase in serum testosterone and reduces general and craniofacial growth in the male Wistar rat. This study aimed to determine whether exogenous testosterone given at an age beyond the normal pubertal peak restores general and craniofacial growth in male, neonatally castrated rats. The design was a randomized, double-controlled, cross-sectional trial. Male Wistar rats were assigned by weighted randomization to be either castrated early after birth (n = 35) or not (n = 15). On day 57, a 1.5-cm Silastic tube with testosterone was implanted in 18 of the castrated rats. On day 70 and day 110, body length, weight, and craniofacial growth were measured together with the weight of the prostate, and blood samples taken. The exogenous testosterone resulted in a significant increase in serum testosterone and prostate weight. All measures of general and craniofacial growth had higher mean values in the non-castrated control group than in the castrated group, while in the testosterone-implant group the mean values lay between these of the castrated animals and the non-castrated controls. Two-way ANOVA indicated a significant effect of the testosterone administration on lower-incisor growth and the size of the total skull vault. PMID- 9730275 TI - A quantitative histometric murine in vivo model of radiation-induced oral mucositis. AB - Gastrointestinal toxicity is a limiting factor in the effectiveness of cancer therapy. This toxicity is most visible in the mouth. There is considerable interest in developing strategies involving growth-factor manipulation of the epithelial stem cells to afford protection to these cells during treatment and/or to speed up the regenerative process following treatment. In order for this to be achieved, studies have to be undertaken in animal systems to demonstrate the proof of principle and determine optimal protocols. Here, a murine model for oral mucositis based on measurements of tissue cellularity at various times after exposure to radiation was used to investigate cytotoxicity. Several sites in the mouth were analysed and the pronounced circadian rhythm in these various epithelial sites determined. The circadian rhythm is important in that it would determine the timing of administration of growth factors. A microscope with an interactive computer was used to define areas of epithelium and lengths of basal layer, within which, and along which, the total number of cell nuclei was determined over a range of times following exposure to 10, 20 and 30 Gy of X rays. For various practical reasons, the ventral surface of the tongue was identified as the most appropriate tissue to analyse. Here, measurements of cellularity reached minimum values between 6 and 8 days following 20 Gy. Labelling of S-phase cells demonstrated foci of regeneration and a burst of proliferative regeneration that commenced at about 5 days and reached peak values at 8 days after irradiation. This burst of regenerative proliferation was coincident with the minimum in tissue cellularity on about day 8. The lower dose of radiation (10 Gy) had minimal effects on cellularity: after the higher dose (30 Gy), there was clearly a more severe level of cellular depletion. This quantitative model of oral mucositis could be used to study the effects of other cytotoxics, including combinations of agents, and the potential role of growth factors to reduce the severity of the cellular depletion and to speed up the kinetics of regeneration. PMID- 9730274 TI - An approach to differentiate between antibacterial and antiadhesive effects of mouthrinses in vivo. AB - An experimental set-up allowing differentiation in vivo between antibacterial and antiadhesive properties of mouthrinses is described. The percentage of vital bacteria (= microbial vitality) and the bacterial counts were microscopically evaluated in saliva and in supragingival dental plaque both collected simultaneously at various times during de novo plaque formation. In a cross-over design, 12 healthy participants refrained from all oral hygiene for four separate periods of 2 x 4 h and 2 x 72 h after having rinsed with either an amine fluoride/stannous fluoride solution (Meridol) or 0.9% NaCl (placebo). Stimulated whole saliva was collected before and after the rinse. Together with whole-saliva samples, representative 4, 24 and 72-h-old plaque samples were separately taken from defined vestibular tooth surfaces that had been either exposed to the mouthrinse (unprotected sites) or temporarily covered with inert plastic films (protected sites) during rinsing. The pooled plaque and saliva were stained with fluorescent dyes to differentiate vital from dead micro-organisms which permitted the estimation of the percentages of vital bacteria. The total bacterial counts were quantified under the darkfield microscope. The Wilcoxon test was used for selected pairwise comparisons (alpha = 0.05). The percentage of vital bacteria in saliva fell significantly from 80-95% to about 50-60% as a result of the antibacterial activity of the test solution. These baseline values and those found in the presence of 4 and 24-h-old plaque were frequently lower than those recorded after the placebo rinse. In comparison to the placebo, microbial vitality was significantly reduced in early supragingival plaque formed on unprotected sites after applying the test solution. The similar total bacterial counts in 4-h-old plaque recorded after the use of the test solution on the unprotected and the protected areas did not point to an antiadhesive effect of the agent. It is concluded that this new experimental set-up allows decoding of the mode of action of a mouthrinse. PMID- 9730276 TI - Development of vimentin filaments in the cells of the articular disc of the rat squamosomandibular joint with age. AB - Age-related changes in the vimentin filaments were studied by immunohistochemistry and electron microscopy. In 2-week-old rats, cells showed only weak labelling for vimentin. A few intermediate filaments but well-developed rough endoplasmic reticulum and Golgi apparatus were observed in the cytoplasm. As the age of the rats advanced from 4-weeks to adulthood, labelling intensity increased and the cytoplasm of articular-disc cells was gradually filled with intermediate filaments surrounding degraded cytoplasmic organelles in the cytosol. Articular-disc cells in adult rats showed intense vimentin labelling and their cytoplasm was filled with closely packed intermediate filaments aligned in alternating layers. This age-related accumulation of vimentin filaments in articular-disc cells may be an adaptive response to compressive forces. PMID- 9730277 TI - The structure and function of yeast xylose (aldose) reductases. AB - Yeast xylose (aldose) reductases are members of the aldo-keto reductase family of enzymes which are widely distributed in a variety of other organisms. In yeasts, these enzymes catalyse the first step of xylose metabolism where xylose is converted to xylitol. In the past 16 years, xylose reductases from yeasts able to ferment or utilize xylose have been isolated and studied mainly because of their importance in xylose bioconversions. In recent years, genes encoding xylose reductases from several yeasts have been cloned and sequenced. A comparison of the primary sequences of yeast xylose reductases with the much better characterized human aldose reductase and human aldehyde reductase reveals that the yeast enzymes are hybrids between aldo-keto reductases and the short chain dehydrogenases/reductases families of enzymes. Why this is so and its evolutionary significance is presently not known. This short review will critically examine the structure and function information that can be gleaned from the sequence comparison. Several interesting questions arise from the sequence comparison and these can provide fruitful areas for further investigations. PMID- 9730278 TI - Multiple regulatory proteins mediate repression and activation by interaction with the yeast Mig1 binding site. AB - A major mediator of glucose repression in yeast is Mig1, a zinc finger protein that binds to a GC-rich recognition sequence found upstream of many glucose repressible genes. Because these Mig1 sites are found upstream of genes under different modes of regulation, we studied regulation of transcription mediated by an isolated Mig1 site placed upstream of a reporter gene under control of UAS(CYC1). The Mig1 site responded appropriately to glucose control and regulatory mutations, including snf1, reg1, cyc8, and tup1, mimicking the behavior of the SUC2 gene. Deletion of the MIG1-coding gene reduced but did not eliminate glucose repression mediated by the Mig1 site. Complete loss of repression was seen in a mig1 mig2 double mutant. When the UAS(CYC1) was replaced by UAS(ADH1) in the reporter plasmid, the Mig1 site activated transcription under most conditions. Mutations of the two Mig1 binding sites in the SUC2 promoter resulted in loss of activation of SUC2 expression. These results suggest the presence of an unknown activator or activators that binds to the Mig1 site. The activator is not any of the proteins previously proposed to bind to this site, including Mig1, Mig2, Msn2, or Msn4. Band shift assays showed that Mig1 is the major protein in yeast cell extracts that binds to the Mig1 site in vitro. This binding is not regulated by glucose or mutations in CYC8 or TUP1. PMID- 9730279 TI - Suppression of sdh1 mutations by the SDH1b gene of Saccharomyces cerevisiae. AB - Transformation of the respiratory-defective mutant (E264/U2) of Saccharomyces cerevisiae with a yeast genomic library yielded two different plasmids capable of restoring the ability of the mutant to grow on non-fermentable substrates. One of the plasmids (pG52/T3) contained SDH1 coding for the flavoprotein subunit of mitochondrial succinate dehydrogenase. The absence of detectable succinate dehydrogenase activity in mitochondria of E264/U2 and the lack of complementation of the mutant by an sdh11null strain indicated a mutation in SDH1. The second plasmid (pG52/T8) had an insert with reading frame (YJL045w) of yeast chromosome X coding for a homologue of SDH1. Subclones containing the SDH1 homologue (SDH1b), restored respiration in E264/U2 indicating that the protein encoded by this gene is functional. The expression of the two genes was compared by assaying the beta-galactosidase activities of yeast transformed with plasmids containing fusions of lacZ to the upstream regions of SDH1 and SDH1b. The 100-500 times lower activity measured in transformants harbouring the SDH1b-lacZ fusion indicates that the isoenzyme encoded by SDH1b is unlikely to play an important role in mitochondrial respiration. This is also supported by the absence of any obvious phenotype in cells with a disrupted copy of SDH1b. PMID- 9730280 TI - Studies of astaxanthin biosynthesis in Xanthophyllomyces dendrorhous (Phaffia rhodozyma). Effect of inhibitors and low temperature. AB - The effect of nicotine and diphenylamine on astaxanthin biosynthesis in Xanthophyllomyces dendrorhous was studied. The effects were analysed under standard and low temperature conditions. It was found that 10 mM-nicotine inhibits the cyclization of lycopene and de novo protein synthesis was not needed to reverse the inhibition. The oxidation of beta-carotene was irreversibly inhibited by 10 microM-diphenylamine while the dehydrogenation of phytoene was reversibly inhibited by 60 microM-diphenylamine. The simultaneous exposure to low temperature (4 degrees C) overcomes the inhibition of beta-carotene oxidation at low diphenylamine concentration. PMID- 9730281 TI - AILV1 gene from the yeast Arxula adeninivorans LS3--a new selective transformation marker. AB - The ILV1 gene of the yeast Arxula adeninivorans LS3 (AILV1) has been cloned from a genomic library, characterized and used as an auxotrophic selection marker for transformation of plasmids into this yeast. One copy of the gene is present in the Arxula genome, comprising 1653 bp and encoding 550 amino acids of the threonine deaminase. The protein sequence is similar (60.55%) to that of the threonine deaminase from Saccharomyces cerevisiae encoded by the gene ILV1. The protein is enzymatically active during the whole period of cultivation, up to 70 h. Maximal activities, as well as protein concentrations of this enzyme, were achieved after cultivation times of 20-36 h. The AILV1 gene is a suitable auxotrophic selection marker in transformation experiments using an Arxula adeninivorans ilv1 mutant and a plasmid containing this gene, which is fused into the 25S rDNA of Arxula adeninivorans. One to three copies of the linearized plasmid were integrated into the 25S rDNA by homologous recombination. Transformants resulting from complementation of the ilv1 mutation can be easily and reproducibly selected and in addition are mitotically stable. Therefore, the described system is preferred to the conventional selection for hygromycin B resistance. PMID- 9730282 TI - Functional analysis of three adjacent open reading frames from the right arm of yeast chromosome XVI. AB - A 7.24 kb genomic DNA fragment from the yeast Saccharomyces cerevisiae chromosome XVI was isolated by complementation of a new temperature-sensitive mutation tsa1. We determined the nucleotide sequence of this fragment located on the right arm of chromosome XVI. Among the three, complete open reading frames: YPR041w, YPR042c and YPR043w contained within this fragment, the gene YPR041w was shown to complement the tsa1 mutation and to correspond to the TIF5 gene encoding an essential protein synthesis initiation translation factor. The YPR042c gene encodes a hypothetical protein of 1075 amino acids containing four putative transmembrane segments and is non-essential for growth. The gene YPR043c encoding the 10 kDa product, highly similar to the human protein L37a from the 60S ribosomal subunit, was found to be essential and a dominant lethal. We conclude that three tightly linked yeast genes are involved in the translation process. PMID- 9730283 TI - A search in the genome of Saccharomyces cerevisiae for genes regulated via stress response elements. AB - Stress response elements (STREs, core consensus AG4 or C4T) have been demonstrated previously to occur in the upstream region of a number of genes responsive to induction by a variety of stress signals. This stress response is mediated by the homologous transcription factors Msn2p and Msn4p, which bind specifically to STREs. Double mutants (msn2 msn4) deficient in these transcription factors have been shown to be hypersensitive to severe stress conditions. To obtain a more representative overview of the set of yeast genes controlled via this regulon, a computer search of the Saccharomyces cerevisiae genome was carried out for genes, which, similar to most known STRE-controlled genes, exhibit at least two STREs in their upstream region. In addition to the great majority of genes previously known to be controlled via STREs, 69 open reading-frames were detected. Expression patterns of a set of these were examined by grid filter hybridization, and 14 genes were examined by Northern analysis. Comparison of the expression patterns of these genes demonstrates that they are all STRE-controlled although their detailed expression patterns differ considerably. PMID- 9730284 TI - The uracil permease of Schizosaccharomyces pombe: a representative of a family of 10 transmembrane helix transporter proteins of yeasts. AB - The uracil permease gene of Schizosaccharomyces pombe was cloned and sequenced. The deduced protein sequence shares strong similarities with five open reading frames from Saccharomyces cerevisiae, namely the uracil permease encoded by the FUR4 gene, the allantoin permease encoded by DAL4, a putative uridine permease (YBL042C) and two unknown ORFs YOR071c and YLR237w. A topological model retaining ten transmembrane helices, based on predictions and on experimental data established for the uracil permease of S. cerevisiae by Galan and coworkers (1996), is discussed for the four closest proteins of this family of transporters. The sequence of the uracil permease gene of S. pombe has been deposited in the EMBL data bank under Accession Number X98696. PMID- 9730285 TI - Current awareness on yeast. PMID- 9730286 TI - HLA expression on immature and mature human germ cells. AB - Human leukocyte antigens (HLA) are membrane-bound glycoproteins encoded by the human major histocompatibility complex located on chromosome 6. They are known to function in immnunologic recognition and, with regard to reproduction, a number of non-immune functions have been proposed. Although the expression patterns of the major histocompatibility antigens have been extensively studied at the maternal fetal interface, there are still controversial reports on the expression of these molecules by human gametes and preimplantation stages. This brief review focuses on recent studies where the expression and distribution of HLA on human spermatogenic cells (spermatogonia, primary and secondary spermatocytes, spermatids, spermatozoa), primary and secondary oocytes, and preimplantation embryos have been investigated. These results, and their possible implications for the fertilization process and further embryonic development, will be presented. PMID- 9730287 TI - Cytokine induced modulation of MHC class I and class II molecules on human cervical epithelial cells. AB - Our purpose was to determine the expression of the major histocompatibility complex (MHC) class I and class II gene products as well as the costimulatory molecules B7-1 and B7-2 on cervical epithelial cells, and to determine to what extent inflammatory cytokines regulate their expression. Immunohistology and flow cytometry techniques were used to identify and quantify MHC class I and class II molecules, and the costimulatory molecules B7.1 and B7.2, on sections and primary epithelial cell cultures of human endo- and ectocervix. MHC class I but not class II molecules were constitutively expressed on tissue sections and primary epithelial cell cultures derived from endo- and ectocervix. Expression of MHC class I and class II was upregulated in vitro by IFN-gamma in a time and dose dependent fashion. The induction of class II expression was more pronounced on ectocervical cells than on endocervical cells. MHC class I but not class II expression was also enhanced by IFN-alpha as well as TNF-alpha. TNF-alpha and TGF beta1 inhibited the IFN-gamma induced MHC class II expression. Expression of the costimulatory molecules B7-1 and B7-2 were not detected in tissue sections or on resting or cytokine-treated cervical epithelial cells in vitro. The present results support the concept that endo- and ectocervical epithelial cells, like their counterparts at other mucosal sites. constitutively express MHC class I molecules and can express MHC class II upon cytokine stimulation, indicating that they are capable of presenting antigens to T-cells. PMID- 9730288 TI - In vitro inhibition of the bioactivity of follicle-stimulating hormone by antisera against a peptide representing part of the FSH-receptor. AB - The aim of the present work was to define an FSH receptor (FSHR) peptide that can induce antibodies that will inhibit the bioactivity of FSH. Therefore, the hFSHR sequence was aligned with that of all other known G-protein coupled receptors. An area with increased sequence homology was identified between the FSH-, LH-, TSH receptors, the C5a receptor and the IL8 receptor. The similarity consists of a richness in acidic (D and E) and hydrophobic (Y and F) residues. In hFSHR the sequence is EDNESSYSRGFDMTYTEFDYDLCNEVVD (amino acid 299-326). Research on both the C5a- and IL8-receptor has indicated that this part is responsible for hormone binding but not for signal transduction. Protamine. an antagonist for both the C5a- and IL8 receptor also inhibited the bioactivities of FSH and LH when tested in a bioassay. This suggests that in the hFSHR this region might also be involved in hormone binding. Specificity of this region towards the diverse ligands all binding to the C5a or to the IL8 receptor might be attributed to differences in the profile of alternating basic and hydrophobic residues. Therefore, the hypothesis was tested as to whether antisera raised against peptides of this FSHR domain would inhibit FSH-bioactivity but not LH-bioactivity. Indeed antisera were found (anti-hFSHR 309-322) that inhibited the biological activity of FSH in a bioassay. These antisera proved to be specific since they did not inhibit the bioactivity of LH. These data suggest that the core sequence (hFSHR 309-322) of the aligned domain of the hFSHR, in analogy to the IL8- and C5a receptors, is involved in hormone binding and ligand specificity. This domain therefore forms a valuable tool in FSH- and FSHR research for scientific and medical purposes. PMID- 9730290 TI - Potential implications of chemokines in reproductive function: an attractive idea. AB - Chemokines are a new family of cytokines specialised in attracting leukocytes, acting in physiological conditions and in pathological processes. A wide variety of cell types in response to exogenous irritants or endogenous mediators of the inflammatory reaction produce them. Pivotal parts of reproductive function are based on inflammatory like processes wherein different leukocytes subsets are recruited and activated to produce paracrine autocrine effects in which cytokines and growth factors are implicated. Since chemokines control leukocyte trafficking and belong to the cytokine superfamily, in this review we analyze the implications of these molecules and related cells in ovulation, embryonic implantation, menstruation, parturition and their role in pathological process such as preterm delivery, endometriosis, ovarian hyperstimulation syndrome and HIV infection. PMID- 9730289 TI - The in vitro antimicrobial capacity of human colostrum against Chlamydia trachomatis. AB - We sought to assess the antimicrobial capacity of human colostrum against Chlamydia trachomatis. a common agent of ophthalmia neonatorum. Colostrum was collected from 13 post-partum females and tested in an in vitro assay of chlamydial growth inhibition using HeLa 229 cells as the host cell line. All samples significantly inhibited chlamydial growth in a dose-response manner. The percent inhibition ranged from 45.3 to 99.0 (mean=88.1+/-4.1). The chlamydial growth inhibition activity of colostrum was found to be: heat- and freezing resistant: more concentrated in colostrum than breast milk; was not attributable to interferon or antibody activity; and, could not be attributed to host cell cytotoxicity. Additionally, chlamydial growth inhibition occurred in < or = 15 min and was effective only when colostrum was incubated with chlamydiae prior to addition to HeLa 229 monolayers. Lastly, centrifugal fractionation of the colostrum yielded similar activity in the lipid pellicle and in the lipid-free supernatant. These results indicate that topically applied colostrum may have efficacy in the prophylaxis of ophthalmia neonatorum of chlamydial etiology in the absence of conventional modalities. PMID- 9730291 TI - Tuberculous pancreatic abscess in an HIV antibody-negative patient: case report and review. AB - Tuberculosis (TB) is most commonly diagnosed as a pulmonary disease; however, haematogenous spread of the organism can cause disease in any organ system. We report the case of a 30-y-old woman, Human Immunodeficiency Virus (HIV) antibody negative, who was diagnosed as having a pancreatic mass on computed tomographic (CT) scans. She underwent a laparotomy and the fluid drained from the mass was culture-positive for Mycobacterium tuberculosis. We review the clinical details of 37 similar cases of pancreatic TB in the literature, where each patient's HIV antibody status is negative or unknown. In this series 3 patients died (1 of these had commenced anti-TB therapy, the others had not) but the remaining 34 responded well to radiological-guided drainage and/or surgical intervention and anti-TB therapy. TB should be considered in the differential diagnosis of a pancreatic mass, especially when associated with epigastric pain or discomfort and weight loss. PMID- 9730292 TI - Moraxella catarrhalis bacteraemia. A report on 3 cases and a review of the literature. AB - Over the last decade, an increase in invasive infections due to Moraxella catarrhalis has been reported. We have analysed 58 cases of bacteraemia due to M. catarrhalis reported in the literature and 3 cases found in Iceland, a total of 61 cases. Patients with bacteraemia could be divided into 3 groups on the basis of host factors. They were either immunocompromised, had underlying respiratory disorders. or were normal hosts. The clinical manifestation of M. catarrhalis bacteraemic infection ranged from a mild febrile illness to a fatal disease. The severity of the clinical picture did not reflect the patients' condition at the time of bacteraemia. The port of entry of the bacteraemia was frequently not elucidated in immunocompromised patients. Patients with a contributory respiratory tract disorder were more likely to develop bacteraemia as a result of a lower respiratory tract infection, whereas bacteraemia in a normal host was more frequently due to an upper respiratory tract infection. The overall prognosis of M. catarrhalis bacteraemia was good, the exception being when it caused endocarditis (5 cases), where mortality rates as high as 80% have been reported. PMID- 9730293 TI - HCV-RNA levels increase during pregnancy in women with chronic hepatitis C. AB - Alanine aminotransferase (ALT) levels decline during pregnancy in women chronically infected with hepatitis C virus (HCV). In order to understand further the underlying mechanisms, we prospectively followed 10 chronically infected women before, during and after pregnancy. ALT levels were analysed together with quantification of serum HCV-RNA using the branched DNA technology. As anticipated, the ALT levels significantly declined late in pregnancy and increased again after delivery. HCV-RNA levels, conversely, significantly increased late in pregnancy and returned to baseline levels 1 y after delivery. These findings suggest the importance of immune mediated mechanisms in controlling the viral replication and contributing to the liver injury in chronic hepatitis C. PMID- 9730294 TI - Immune thrombocytopenic purpura after recombinant hepatitis B vaccine: retrospective study of seven cases. AB - Recombinant hepatitis B vaccine is usually well tolerated. Clinical and laboratory test manifestations with immunologic mechanisms have nonetheless been described following use of this vaccine. We retrospectively report 7 cases of thrombocytopenia occurring within 3 months (7 weeks on the average) of 1 or following injections of recombinant hepatitis B vaccine. Four boys and 3 girls, average age 12 y, were involved. Three had a history of immune thrombocytopenic purpura. Four had haemorrhagic manifestations. The haemogram showed thrombocytopenia (24 x 10(9)/l on the average) without alterations of the other lines. Infectious and immune aetiologies were excluded in all cases. The course varied after treatment by corticosteroids, high-dose intravenous immunoglobulin, or both. After describing the different manifestations subsequent to recombinant hepatitis B vaccination, we discuss post-vaccinal thrombocytopenias (vaccines in question, mechanisms) and the reality of this entity. PMID- 9730295 TI - Prolonged nosocomial outbreak of hepatitis A arising from an alcoholic with pneumonia. AB - From April to June 1996, an outbreak of hepatitis A virus (HAV) infection affecting 15 nurses, patients and household contacts occurred in the Department of Internal Medicine at Aker University Hospital, Oslo. The index case was a homeless alcoholic who was hospitalized in March 1996 with pneumonia while simultaneously incubating HAV infection. Four secondary cases were infected by the index case, while another 10 cases were caused by a continuous spread of infection within the department during the following months. Sequence of the VP1/P2A junction of HAV was obtained from 9 patients, including the index case, and all sequences were identical to each other. Mass vaccination of hospital employees with a formalin-inactivated HAV-vaccine took place in late May, and following this the outbreak stopped. Several factors probably combined to account for this unusual outbreak, e.g. an index case unsuspected of incubating with HAV infection, and a low prevalence rate of protective antibodies to HAV among the hospital staff. PMID- 9730296 TI - Comparison of cytochemical staining, immunofluorescence and PCR for diagnosis of pneumocystis carinii on sputum samples. AB - Detection of P. carinii has increased with the use of polymerase chain reaction (PCR), particularly in sputum samples. In this study, sputum samples obtained from 30 immunosuppressed patients with respiratory symptoms (12 HIV-infected) were tested by standard cytochemical staining (Giemsa and methenamine silver), immunofluorescence (IF) staining and PCR for detection of P. carinii and the results were compared. Pneumocystis carinii was detected in 4, 8 and 13 sputum samples by cytological staining, IF test and PCR, respectively. Specific amplification bands were obtained in all sputum samples that were positive by both other tests. All tests gave negative results in sputum samples obtained from 5 HIV-infected asymptomatic patients and 22 non-immunosuppressed tuberculosis patients. Our observations suggest that PCR results were well correlated with P. carinii pneumonia (PCP), especially in non-HIV-infected patients. However, PCR positivity obtained in HIV-infected patients could be misleading in the diagnosis of PCP without careful clinical evaluation. Positive results obtained by Giemsa staining or IF test confirm diagnosis of PCP authoritatively. As a result, we suggest testing sputum samples by both PCR and IF techniques for detection of P. carinii. PMID- 9730298 TI - Serum sCD23 in patients with lepromatous and tuberculoid leprosy. AB - Tuberculoid and lepromatous leprosy (TL and LL) manifest exaggerated Th1 and Th2 type immunity, respectively. Serum soluble CD23, which is regulated by the stimulatory action of IL4 and inhibitory action of IFNgamma, was significantly elevated in LL relative to TL and healthy controls. These results confirm the state of cellular and humoral immunity in TL and LL. PMID- 9730297 TI - Rapid detection of rifampin-resistant Mycobacterium tuberculosis by sequencing and line probe assay. AB - Resistance to rifampin (RMP) is associated with mutations in the rpoB gene. Disk elution method, direct DNA sequencing and line probe assay were compared in rapid detection of rpoB mutations from 30 Mycobacterium tuberculosis isolates. Concordance between LiPA and culture was 93.3%, whereas sequencing yielded 100% concordance with culture. These results indicate that line probe assay is nearly as sensitive a method as sequencing in rapid detection of RMP-resistance of M. tuberculosis isolates, and can be applied in laboratories working with standard PCR equipment. PMID- 9730299 TI - Outbreak of Candida albicans fungaemia in a neonatal intensive care unit. AB - During a 4-month period, 9 premature infants hospitalized in a neonatal intensive care unit (NICU) developed Candida albicans fungaemia. All 9 infants received antifungal agents. Fluconazole was administered in 7 patients and successfully eradicated this organism in 6 with no adverse effects. For epidemiological investigation, 64 environmental specimens and hand-washings of all 54 staff members involved in the NICU were examined for the presence of this organism. No C. albicans could be identified from environmental sources, while the hand washing of 1 nurse was C. albicans-positive. Two genotyping methods, including electrophoretic karyotyping using contour-clamped homogeneous electric field gel electrophoresis and polymerase chain reaction-based direct sequencing of rRNA gene, were used in the analysis of the isolates recovered from blood cultures of the infants, the hand-washing of the nurse and 7 control isolates. Both methods yielded comparable results and revealed that all 13 isolates from infected infants as well as the isolate from hand washing of the nurse were of the same genotype while the control isolates were distinct. These results suggest that the outbreak of C. albicans fungaemia was caused by a particular strain and possibly via cross-infection. In addition, we showed that fluconazole seemed to be safe and effective in treating C. albicans fungaemia in neonates, although the data were limited. PMID- 9730300 TI - Successful treatment of disseminated Mycobacterium simiae infection in AIDS patients. AB - Mycobacterium simiae is rarely isolated in clinical settings of non-HIV-infected patients. Isolation of M. simiae from clinical specimens in clusters has been limited to some parts of the world that include Israel, Cuba and the southern USA, mainly Texas. Only 8 patients with HIV and disseminated M. simiae infection have been previously described in the English literature. Successful treatment of disseminated M. simiae infection has never been reported and 6 of the cases have died within 8 months of diagnosis. We reviewed retrospectively the medical records of HIV-infected patients who had positive blood or bone marrow cultures for M. simiae at the Sheba Medical Center, Tel-Hashomer, Israel, between January 1992 and December 1996. A case of disseminated M. simiae infection was defined as isolation of M. simiae in blood or bone marrow culture in an HIV-infected patient with a compatible, otherwise unexplained, systemic disease. Mycobacterium simiae was isolated in blood and/or bone marrow cultures from 3 HIV-infected patients during the last 5 y. We describe the first successful treatment in AIDS patients with disseminated M. simiae infection. The patients are alive and well 20 months after instituting a combination of 3 antimycobacterial agents, clarithromycin, ethambutol and ciprofloxacin and intensive antiretroviral therapy. PMID- 9730301 TI - Vertebral osteomyelitis at a Norwegian university hospital 1987-97: clinical features, laboratory findings and outcome. AB - Altogether 40 patients aged 13-91 y (average 58 y) with vertebral osteomyelitis were treated at the Bergen University Hospital between July 1987 and June 1997. All patients presented with back pain, 33 (83%) had vertebral tenderness, and 26 (65%) patients were febrile. The duration of symptoms before diagnosis was < 3 weeks in 13 patients, and from 3 to 16 weeks in the remaining 27 patients. C reactive protein (CRP) level and erythrocyte sedimentation rate (ESR) were elevated in 39 and 38 patients, respectively. Staphylococcus aureus was the most frequent cause of osteomyelitis followed by Streptococcus spp., Escherichia coli and Mycobacterium tuberculosis. Magnetic resonance imaging was superior to other radiological methods and demonstrated changes consistent with osteomyelitis in all 23 patients examined with this method. 35 patients survived. 18/35 surviving patients had pareses and 17 underwent surgery with drainage of abscesses or laminectomy. All 35 patients made a good recovery and only 3 patients experienced permanent pareses. The diagnosis of vertebral osteomyelitis is easily missed, and treatment is often delayed, particularly in the elderly in whom signs of sepsis may not manifest. However, persisting localized pain and tenderness over the spine together with elevated CRP and ESR should prompt the physician to consider vertebral osteomyelitis. Fever and leukocytosis may support the diagnosis, but may not always be present. PMID- 9730302 TI - Comparison of risk factors and outcome in patients with Enterococcus faecalis vs Enterococcus faecium bacteraemia. AB - The purpose of our study was to determine retrospectively the risk factors for the acquisition of Enterococcus faecalis vs E. faecium bacteraemia, as well as the clinical outcomes of these patients. 62 patients with Enterococcus faecalis bacteraemia were compared to 31 patients with E. faecium bacteraemia. Haematologic malignancies, neutropenia, high-risk source and previous use of aminoglycosides, carbapenems, cephalosporins and clindamycin were significantly associated with E. faecium bacteraemia. Instead, urinary catheterization was found to be related to Enterococcus faecalis bacteraemia. The mortality rates within 7 d and 30 d were 13% and 27%, respectively, in patients with E. faecalis bacteraemia and 6% and 29%, respectively, in patients with E. faecium bacteraemia. There was no difference in mortality between E. faecalis and E. faecium bacteraemia, nor was there a difference in seriousness of disease at the time of bacteraemia. In the subgroups of patients with monomicrobial or clinically significant E. faecalis vs E. faecium bacteraemia, the mortality rates were similar to the results of all subjects. Our results do not support the theory that E. faecium would be a more virulent organism than E. faecalis. PMID- 9730303 TI - Age-related prevalence of Shigella and Salmonella antibodies and their association with diarrhoeal diseases in Peruvian children. AB - Shigella and Salmonella antibodies in relation to diarrhoea were studied in a cohort of 413 children between 2 and 27 months of age in peri-urban Lima, Peru. Blood samples were obtained at 2, 3 and 12 months of age. Antibody titres against lipopolysaccharide from Shigella flexneri serotype Y, Shigella dysenteriae serotype 1, Shigella sonnei, Salmonella serogroups AO, BO, DO, and Shigella Ipa and Salmonella typhi Vi antigens were measured by enzyme immunoassay. IgG titres against S. flexneri and Shigella Ipa were higher at 2 than at 3 or 12 months of age (p=0.001), while the changes in IgG titres against S. dysenteriae, S. sonnei and Salmonella were not pronounced. IgA and IgM titres against S. flexneri, Shigella Ipa, S. dysenteriae, S. sonnei and Salmonella were significantly higher at 12 than at 2 or 3 months of age (p=0.001). Stool samples were obtained from children in 64% of all diarrhoeal episodes. Shigella spp. were isolated from 20% of the children during the first 2 y of life and Salmonella in 3%. Most isolates were from children at 13-24 months of age (78%). IgG antibodies at 12 months of age did not protect against shigellosis during the second year of life. PMID- 9730304 TI - Epidemiology of invasive Haemophilus influenzae type b infections among children in Greece before the introduction of immunization. AB - We prospectively examined the epidemiology of invasive Haemophilus influenzae type b (Hib) infections among children under 5 y of age in the Greater Athens area before the introduction of immunization. The annual incidence of systemic Hib infections was 12/100000. Meningitis was the most common clinical entity and accounted for 69% of the cases. In the prevaccine era, the incidence of systemic Hib disease, particularly that of meningitis, was much lower in Greece compared to rates reported from Northern and Central Europe. PMID- 9730305 TI - Endotoxaemic liver injury in a porcine model--relation to tumour necrosis factor alpha release and survival. AB - The effects of a continuous infusion of E. coli endotoxin on liver blood chemistry, plasma TNFalpha and blood pressure were evaluated in relation to survival in 18 anaesthetized pigs. The endotoxin infusion cause both unconjugated and conjugated bilirubin to increase (both p < 0.001), as compared to 6 saline infused controls. Four pigs died during the endotoxin infusion. The plasma levels of TNFalpha of these 4 non-survivors were higher compared to TNFalpha of the 14 surviving endotoxaemic pigs (p < 0.05). The increase in plasma bilirubin of the non-survivors was less expressed (p < 0.05), and their arterial blood pressure and base excess deteriorated (both p < 0.001) compared to surviving endotoxaemic pigs. A decreased blood pressure explains both the reduced 'hepatic washout' of bilirubin and visceral ischaemia, reflected by the development of metabolic acidosis. This porcine model may be suitable for studying endotoxaemic liver injury. PMID- 9730306 TI - Immunoserologic evidence of Human Granulocytic Ehrlichiosis in Danish patients with Lyme neuroborreliosis. AB - Human Granulocytic Ehrlichiosis (HGE) is a recently described human illness in the US which manifests as fever, myalgia and headache combined with pancytopenia and elevated concentrations of hepatic transaminases. Genetic analyses indicate that the agent of HGE appears to be an Ehrlichia species that is closely related to E. equi and E. phagocytophila. Ixodes dammini and I. scapularis were identified as potential vectors of HGE. Ixodes ticks are also the vector of Borrelia burgdorferi, the agent of Lyme borreliosis. The presence of antibodies against Ehrlichia in 132 sera from Danish patients with definite Lyme neuroborreliosis were examined in order to provide immunoserologic evidence of this infection in Denmark. Patients with Lyme neuroborreliosis were chosen as a test cohort, as these patients had been infested by a tick sufficient for transmission of B. burgdorferi. All had cerebrospinal fluid lymphocytic pleocytosis. As controls, serum samples from 50 healthy Danish blood donors were included. Of the 132 patients with Lyme neuroborreliosis, 5 (3.8%) reacted with the E. equi antigen substrate at titres 1:128. None of the blood donors were found seropositive for E. equi. At least 2 of the patients found seropositive for HGE constituted probable cases of HGE with E. equi antibody titres of at least 80 combined with fever, headache and myalgias. However, in no cases were we able to detect the presence of the HGE agent in the serum by PCR. We conclude that human exposure to granulocytic Ehrlichiae species may also occur in Europe, although further studies will be necessary to document active infection with these potential pathogens. PMID- 9730307 TI - Seroepidemiology of Mycoplasma pneumoniae infections in Iceland 1987-96. AB - Mycoplasma pneumoniae is a common cause of respiratory tract infections in humans. The aim of the present study was to analyse the seroepidemiology of M. pneumoniae infections in Iceland during a 10-y period. A retrospective analysis of all serological diagnosis of M. pneumoniae infections at the Department of Medical Virology, National University Hospital in Reykjavik was performed. A total of 13,201 test results from 1987 to 1996 were reviewed and altogether 762 patients were found to have raised M. pneumoniae antibody titres, using a conventional complement fixation assay. Infections were most common amongst young people (< or=16 y) but a second peak in incidence was observed around the age of 35 y. Significant annual (p < 0.0001) and seasonal variations (p=0.0003) were observed; M. pneumoniae infections were most common during the winter period. Three major outbreaks with intervals of three to five years were observed during the observation period. Patients diagnosed during these outbreaks had higher M. pneumoniae titres than those found when infections were less frequent (p=0.0017). Furthermore, the middle aged and elderly patients (> 50 y) had significantly lower M. pneumoniae titres than younger patients (p=0.0014). The findings of this study show that M. pneumoniae infections have definite annual and seasonal variations and also confirm previous studies showing community outbreaks of M. pneumoniae infections every 3-5 y. PMID- 9730308 TI - Mycoplasma contamination of Chlamydia pneumoniae isolates. AB - We examined 6 C. pneumonia isolates from The American Type Culture Collection (ATCC) and 2 Finnish isolates for Mycoplasma contamination. Three of the ATCC isolates and both of the Finnish isolates were Mycoplasma-contaminated. The contaminants were characterized by means of growth in BEa and BEg media, immunoblotting, polymerase chain reaction and pulsed field gel electrophoresis. Two of the 6 ATCC isolates [ATCC VR1355 (TWAR strain 2043) and ATCC VR1356 (TWAR strain 2023)] were infected with Mycoplasma hominis and 1 isolate [ATCC VR2282 (TWAR strain TW183)] was contaminated with both Mycoplasma hominis and Mycoplasma orale, whereas 3 of the ATCC isolates [ATCC VR1310, ATCC VR1360 (TWAR strain CM 1) and ATCC 53592 (TWAR strain AR39)] were not contaminated. The Finnish C. pneumoniae isolates Kajaani 6 and Parola were found to be contaminated with M. hominis and M. orale, respectively. The contamination of C. pneumoniae stock cultures, frequently used in the microimmunofluorescence test, with human pathogens, could pose a serious problem in C. pneumoniae serology. PMID- 9730309 TI - Fatal necrotizing fasciitis caused by a toothpick injury. AB - Necrotizing fasciitis is a severe life-threatening infection. The portal of entry is usually a site of disruption of the skin barrier. We report a case of fatal necrotizing fasciitis caused by an accidental toothpick injury--a unique injury mechanism not reported this far to cause necrotizing fasciitis. Although toothpick injuries are usually regarded as trivial, it should be kept in mind that they have the potential to cause such a lethal infection. PMID- 9730310 TI - Simultaneous Aspergillus fischeri and Herpes simplex pneumonia in a patient with multiple myeloma. AB - A patient with light chain myeloma complicated by simultaneous Herpes simplex and Aspergillus fischeri pneumonia is presented. Microscopic examination of her bronchoalveolar specimen showed bronchial cells with cytopathic effects and numerous cleistotheca, the sexual reproductive structures of Aspergillus. Culture was positive for Herpes simplex virus and Aspergillus fischeri. The initial partial response to amphotericin B followed by complete clinical response with addition of intravenous acyclovir emphasized the importance of recognition of simultaneous infection by these 2 pathogens. This is the first report of identifying cleistotheca in the bronchoalveolar lavage specimen. PMID- 9730311 TI - Necrotizing fasciitis due to Vibrio alginolyticus following an injury inflicted by a stingray. AB - We describe the case of a patient with necrotizing fasciitis due to Vibrio alginolyticus in a patient with cirrhosis following an injury inflicted by a stingray. The patient was successfully treated with aggressive surgical debridement and a combination of ciprofloxacin and amoxicillin-clavulanate. Cases of invasive V. alginolyticus reported in the literature were reviewed. PMID- 9730312 TI - Limb-threatening necrotizing alternariosis salvaged by adjunctive hyperbaric oxygen therapy. AB - We describe, to our knowledge, the first case of limb-threatening necrotizing alternariosis whose limbs were successfully salvaged by adjunctive hyperbaric oxygen therapy (HBO2). This 58-y-old patient was immunocompromised with both diabetes and Cushing's syndrome. She suffered from necrotizing soft tissue infection of both legs caused by Alternaria. It was impossible to halt the progression of the invasive infection with standard anti-fungal treatment and aggressive surgical debridement. After the use of HBO2, the wound was stabilized and eventually healed. Adjunctive HBO2 in this case has demonstrated its role in the treatment of complicating necrotizing soft tissue infection caused by invasive fungal infection. The possible mechanisms may be the potentiation of immune responses and the enhancement of fibroblast proliferation. PMID- 9730313 TI - Acute meningococcal epiglottitis and septicaemia in a 65-y-old man. AB - We report a case of acute meningococcal epiglottitis in a 65-y-old man. He was noted to have stridor of acute onset. We highlight the importance of the diagnosis of acute epiglottitis, early establishment of an airway and appropriate antibiotic therapy. This case report mainly concerns the association of unusual pathogen Neisseria meningitidis and adult acute epiglottitis. PMID- 9730314 TI - Pharyngolaryngitis caused by Neisseria meningitidis. AB - Neisseria meningitidis is a causative agent of life-threatening cases of meningitis and sepsis, but it can also cause mild and self-limiting bacteraemia. Patients with N. meningitidis sepsis or meningitis often describe signs of upper respiratory tract infection before the onset of invasive disease. Viral respiratory infections have been associated with invasive meningococcal diseases and they may contribute to these prodromal symptoms. N. meningitidis can be cultivated from the throats of asymptomatic carriers and it likely enters the circulation through the upper respiratory tract. However, it is unclear whether N. meningitidis can cause simple pharyngitis. Here we describe a case of acute fulminant pharyngolaryngitis caused by N. meningitidis as verified by positive blood cultures. PMID- 9730315 TI - Septicaemia with Neisseria elongata ssp. nitroreducens in a patient with hypertrophic obstructive cardiomyopathia. AB - Neisseria elongata ssp. nitroreducens, a commensal of the human upper respiratory tract, is a newly recognized cause of endocarditis. We report the isolation of the organism from blood cultures of a 30-y-old man with hypertrophic obstructive cardiomyopathy. The patient was successfully treated with benzylpenicillin and netilmicin in combination, followed by ceftriaxone and amoxicillin. PMID- 9730316 TI - Congenital toxoplasma gondii infection diagnosed by PCR amplification of peripheral mononuclear blood cells from a child and mother. AB - A case of congenital toxoplasmosis is presented, where diagnosis by PCR amplification of Toxoplasma gondii DNA from peripheral blood led to early treatment of the infant and seemingly normal brain development despite the presence of intracranial calcifications at birth. The mother, who experienced a subclinical infection during pregnancy, was PCR-positive for toxoplasma DNA in a sample of peripheral blood drawn after delivery. PMID- 9730317 TI - Thrombosis of the ulnar veins--an unusual manifestation of Loa loa filariasis. AB - A 49-y-old male, who had travelled in Central Africa, was admitted to hospital with oedema of the right forearm. He was diagnosed as having thrombosis of the ulnar veins. Subsequently eosinophilia and positive serology for filariae established the aetiologic agent as Loa loa. The patient received antithrombotic and antiparasitic therapy. This is, to our knowledge, the first reported case of thrombosis of the ulnar veins associated with Loa loa. PMID- 9730318 TI - Relapse of erysipelas following treatment with prednisolone or placebo in addition to antibiotics: a 1-year follow-up. AB - Of 112 patients with erysipelas, who were randomized to treatment with 8 d of either prednisolone or placebo in addition to antibiotics, 103 were followed-up for 12 months after they had been cured. The results of the period from 3 weeks up to 1 y after the day of cure are presented. 52 patients came from the prednisolone group and 6 of them had further episodes of erysipelas, whereas 13/51 patients from the placebo group relapsed. The difference is not statistically significant. PMID- 9730319 TI - Tyrosinase autoactivation and the problem of the lag period. AB - Evidence is presented for the binding of the quinone oxidation product of the monohydric phenol substrate, 4-hydroxyanisole, to mushroom tyrosinase. Column chromatography and SDS-PAGE separation showed labelling of the enzyme when incubated with 14C ring-labelled 4-hydroxyanisole. It is proposed that covalent binding to the enzyme and other proteins is through reaction of accessible nucleophilic groups, including thiols and amino groups, with the anisylquinone. This reductive addition enables the indirect generation of the catecholic substrate, which acts as an electron donor for the bicupric active site of met tyrosinase and explains the lag kinetics of tyrosinase oxidation of non-cyclizing substrates. The effects of diluting the enzyme or the addition of amino acids on the lag period was consistent with a mechanism involving indirect generation of the dihydric phenol, which acts as the met-enzyme-recruiting substrate. PMID- 9730320 TI - Human melanoma cell lines show little relationship between expression of pigmentation genes and pigmentary behaviour in vitro. AB - Several laboratories are pursuing the question of whether the expression of pigment genes can be used as a useful marker for tumour progression. However, many melanoma tumours are amelanotic in vivo. The purpose of this study was to examine the relationship between the expression of tyrosinase-related genes [tyrosinase, tyrosinase-related protein-1 (TRP-1) and tyrosinase-related protein 2 (TRP-2)] and pigmentation of melanoma cells. Fourteen cutaneous melanoma cell lines were examined for visible pigment, melanin content, and dopa oxidase activity and findings were related to the previously determined expression of the three tyrosinase-related genes in these cells in culture. Four of the cell lines were also stimulated with alpha-MSH, isobutylmethylxanthine, and forskolin to examine the relationship between induced pigmentation and upregulation of pigmentation genes. There was no simple correlation between pigmentation gene expression and dopa oxidase activity or total melanin content of the 14 melanoma cell lines in culture. In the majority of cells, there was no appreciable pigment, whereas, in contrast, half of the cells showed significant dopa oxidase activity. Upregulation of dopa oxidase activity was achieved by alpha-MSH in two out of four cell lines examined in detail and with IBMX in three out of four of these cell lines. IBMX increased tyrosinase gene expression in all four cell lines; alpha-MSH was without effect; and TRP-1 and TRP-2 expression were largely unaffected by IBMX or alpha-MSH. Modest changes in morphology were noted in response to IBMX. Overall, however, human melanoma cell lines were, with two exceptions, amelanotic in culture despite the fact that 10 out of the 14 lines expressed tyrosinase-related genes. We conclude that measurable pigmentation is not a necessary consequence of the expression of pigmentation genes. An implication of this work is that amelanotic tumours in vivo may nevertheless be positive for tyrosinase-related genes. PMID- 9730321 TI - Lincomycin abrogates dexamethasone-enhanced melanogenesis in B16 melanoma cells. AB - The effects of lincosamide, and interference between the effects of glucocorticoid and lincosamide, on melanogenesis were determined in B16 melanoma cells. Cells were treated for 4 days with lincomycin (LM) and/or dexamethasone (DX) at equimolar concentrations ranging from 10(-9) M to 10(-5) M, or at various concentrations of DX with 10(-6) M LM. Effects on proliferation, tyrosinase activity, melanin biosynthesis, and levels of mRNA for tyrosinase, tyrosinase related protein 1 (TRP1), and tyrosinase-related protein 2 (TRP2) were examined. Treatment with LM or LM + DX stimulated proliferation of melanoma cells with minimal cytotoxicity, while DX did not influence cell proliferation either alone or in combination with LM. Treatment with LM alone increased tyrosinase activity slightly and reduced melanin content in a dose-dependent manner. However, LM counteracted the pronounced increase in tyrosinase elicited by DX and also abrogated the dose-dependent increase in melanin content elicited by DX. Treatment with LM alone did not affect mRNA levels for tyrosinase, TRP1, or TRP2. Furthermore, LM abrogated the DX-induced up-regulation of mRNAs for tyrosinase and the down-regulation of TRP1 mRNA. These results suggest that LM inhibits melanogenesis post-transcriptionally and abrogates glucocorticoid-induced melanogenesis at the transcriptional level in B16 melanoma cells. PMID- 9730322 TI - Dermatoscopy and high frequency sonography: two useful non-invasive methods to increase preoperative diagnostic accuracy in pigmented skin lesions. AB - Dermatoscopy and high frequency sonography have recently been combined to increase diagnostic preoperative accuracy in the treatment of pigmented skin lesions. In this monocentric study 80 patients with pigmented skin lesions were evaluated clinically, by dermatoscopy, and 20 MHz-sonography followed by dermatohistopathological evaluation; 39 malignant melanomas, 37 common nevi, 3 dysplastic nevi, and 1 nevus Spitz were diagnosed histologically. In 72 of the 80 cases (91.3%) dermatoscopical diagnoses were confirmed by histopathology, compared to only 79% correct clinical diagnoses. For the mere clinical diagnosis of melanoma sensitivity was 79%, specificity was 78% and diagnostic accuracy was 65%. All diagnostic values increased by dermatoscopy: sensitivity reached 90%, specificity was 93%, and diagnostic accuracy was 83%. In order to determine tumor thickness preoperatively tumor thickness was measured by 20 MHz sonography. The correlation of tumor thickness between histometric and sonographic results was determined for nevi (r = 0.93) and melanoma (r = 0.95); 74.3% of melanomas were diagnosed correctly within an 0.2 mm range. Regarding the clinical important limit of 1 mm tumor thickness, 87.2% were diagnosed in accordance with histometric evaluation. An increase of 18% in diagnostic accuracy by dermatoscopy and 87.2% of correctly diagnosed cases of tumor thickness of malignant melanoma by high frequency sonography clearly demonstrate that these methods should be considered standard procedures in the diagnosis of pigmented skin lesions and will facilitate the decision on necessary surgical treatment. PMID- 9730323 TI - Isolation, cloning, and sequencing of porcine agouti exon 2 (PorAex2). AB - In order to isolate, clone, and sequence agouti exon 2 of the pig (Yorkshire), we used an interspecific hybridization strategy. Primers from the 5' and 3' borders of the known human agouti exon 2 sequence were used to amplify (PCR) pig agouti exon 2. Following Southern blotting using a human exon 2 internal primer to authenticate that our PCR amplified product was truly pig exon 2 (PorAex2), the fragment was cloned and sequenced. PorAex2 exhibits 79.1 and 75.7% DNA sequence and 85 and 74% deduced amino acid sequence homologies with human and mouse agouti exon 2 and agouti protein, respectively. With the isolation of PorAex2, we can now map, sequence, and clarify the modus operandi of the porcine agouti gene. The GenBank Accession number of PorAex 2 is AF018166. PMID- 9730324 TI - Three-dimensional CT measurement of adult acetabular dysplasia: technique, preliminary results in normal subjects, and potential applications. AB - OBJECTIVE: To assess a three-dimensional computed tomography (3DCT) technique for measurement of acetabular coverage in adults. DESIGN: We used 3DCT to define the geometric centre of the femoral head and to measure centre-edge angles (CEAs) at 10 degrees rotational increments around the acetabular rim. The means, ranges, standard deviations and 95% confidence intervals for the CEAs at the various rotational increments were determined. Inter- and intra-observer variability was measured. The normal values are compared with two example cases of acetabular dysplasia. PATIENTS: The normal hips of 15 subjects aged 1949 years (mean 34.2 years) were measured. RESULTS: The 3DCT measurements are reproducible (mean difference interobserver, 1.7 degrees - 7.9 degrees; mean difference intra observer, 0.6 degrees-6.9 degrees). Mean normal CEA at the lateral rim was 33 degrees with a 95% confidence interval of 23 degrees - 43 degrees. Mean normal CEAs at 10 rotational increments from anterior to posterior rim were determined, and graphed as a 'normal curve'. CONCLUSION: This new 3DCT method of assessing acetabular dysplasia is simple, reproducible, and applicable to diagnosis, quantification and surgical planning for adult acetabular dysplasia patients. PMID- 9730325 TI - Imaging features of subcutaneous sarcoidosis. AB - OBJECTIVE: This report describes subcutaneous sarcoidosis, focusing on the radiological and magnetic resonance (MR) features of the disease. DESIGN AND PATIENTS: The cases of four patients (one male and three female, age range 36-75 years) who had subcutaneous sarcoidosis with no other organs affected were reviewed. Lesions were nodular in two cases, and in the other two were diffuse. RESULTS: Computed tomography (CT) demonstrated a well-defined, homogeneous, and enhanced lesion in the nodular cases. However, in the diffuse cases, CT showed a heterogeneous, honeycomb-like appearance and little enhancement. Angiography showed a fine stain in the arterial phase. MR imaging of the nodular lesions was homogeneous with a signal intensity similar to muscle on T1-weighted images but heterogeneous with a higher signal than muscle on T2-weighted images. Diffuse lesions showed a striped or mesh pattern with intermediate signal intensity on both T1- and T2-weighted images. Contrast-enhanced MR images showed slight enhancement. CONCLUSIONS: Subcutaneous sarcoidosis should be considered in the differential diagnosis when a patient presents with the radiological and MR features described. PMID- 9730327 TI - Chondrocalcinosis of the hyaline cartilage of the knee: MRI manifestations. AB - PURPOSE: To determine the ability of MRI to detect the presence of crystals of calcium pyrophosphate in the articular cartilage of the knee. DESIGN AND PATIENTS: The MR studies of 12 knees (11 cases) were reviewed retrospectively and correlated with radiographs (12 cases) and the findings at arthroscopy (2 cases) and surgery (1 case). A total of 72 articular surfaces were evaluated. Radiographic, surgical or arthroscopic demonstration of chondrocalcinosis was used as the gold standard. Additionally, two fragments of the knee of a patient who underwent total knee replacement and demonstrated extensive chondrocalcinosis were studied with radiography and MRI using spin-echo T1-, T2- and proton-density weighted images as well as two- and three-dimensional fat saturation (2D and 3D Fat Sat) gradient recalled echo (GRE) and STIR sequences. RESULTS: MRI revealed multiple hypointense foci within the articular cartilage in 34 articular surfaces, better shown on 2D and 3D GRE sequences. Radiographs showed 12 articular surfaces with chondrocalcinosis. In three cases with arthroscopic or surgical correlation, MRI demonstrated more diffuse involvement of the articular cartilage than did the radiographs. The 3D Fat Sat GRE sequences were the best for demonstrating articular calcification in vitro. In no case was meniscal calcification identified with MRI. Hyperintense halos around some of the calcifications were seen on the MR images. CONCLUSION: MRI can depict articular cartilage calcification as hypointense foci using GRE techniques. Differential diagnosis includes loose bodies, post-surgical changes, marginal osteophytes and hemosiderin deposition. PMID- 9730326 TI - Shoulder MRI after impingement test injection. AB - OBJECTIVE: To determine how long injected fluid from an impingement test remains in the bursa or adjacent soft tissues after an injection. DESIGN AND PATIENTS: Six patients prospectively underwent MRI of the shoulder immediately before and after an impingement test injection, and at 3 days, 2 weeks and 4 weeks later. MR images were evaluated and graded for fluid distribution within the bursa and adjacent soft tissues. The rotator cuff was evaluated for signal abnormalities related to the injection. RESULTS AND CONCLUSION: Three days after the injection, the soft tissue fluid had returned to pre-injection levels or less in five of the six patients. No patients showed rotator cuff signal abnormalities related to the impingement test injection. We recommend a delay of 3 days before a shoulder MR examination after an injection has been performed, to avoid misinterpretations. PMID- 9730328 TI - Posterior interosseous nerve palsy caused by parosteal lipoma of proximal radius. AB - Lipomas are common benign soft tissue tumors which tend to be indolent, and symptoms caused by nerve compression are unusual. However, a parosteal lipoma, occurring adjacent to the proximal radius may easily cause paralysis of the posterior interosseous nerve because of a specific anatomical relationship of these structures in that area. Two cases of parosteal lipoma of the proximal radius causing paralysis of the posterior interosseous nerve are reported. CT and MR imaging demonstrate the characteristic fatty mass around the radius and are specific in making the diagnosis. Surgical excision should be promptly performed to ensure optimal recovery from the nerve paralysis. PMID- 9730329 TI - Measurements of vertebral shape by radiographic morphometry: sex differences and relationships with vertebral level and lumbar lordosis. AB - OBJECTIVE: To examine sex-related and vertebral-level-specific differences in vertebral shape and to investigate the relationships between the lumbar lordosis angle and vertebral morphology. DESIGN AND PATIENTS: Lateral thoracic and lumbar spine radiographs were obtained with a standardized protocol in 142 healthy men and 198 healthy women over 50 years old. Anterior (Ha), central (Hc) and posterior (Hp) heights of each vertebra from T4 to L4 were measured using a digitizing technique, and the Ha/Hp and Hc/Hp ratios were calculated. The lumbar lordosis angle was measured on the lateral lumbar spine radiographs. RESULTS: Ha/Hp and Hc/Hp ratios were smaller in men than women by 1.8% and 0.7%, respectively, and these ratios varied with vertebral level. Significant correlations were found between vertebral shape and the lumbar lordosis angle. CONCLUSIONS: These results demonstrate that vertebral shape varies significantly with sex, vertebral level and lumbar lordosis angle. Awareness of these relationships may help prevent misdiagnosis in clinical vertebral morphometry. PMID- 9730330 TI - Synovial chondromatosis in a lumbar apophyseal joint. AB - A 31-year-old woman presented with painful swelling in the right paravertebral region that had been present for 2 years. Radiography and CT revealed an area of increased density due to multiple calcifications localized at the fourth lumbar vertebra. Histological examination revealed that the lesion consisted of nodules of hyaline cartilage, with focal areas of calcification, growing within synovial tissue. PMID- 9730331 TI - Congenital maldevelopment of intervertebral disc simulating a neurofibroma. AB - Intervertebral disc herniation is a common cause of radiculopathy and myelopathy in adulthood. It is an uncommon tumor mimic. We report on an extradural disc associated with an osseous defect ostensibly caused by pressure erosion and appearing as a neural tumor. It showed homogeneous enhancement on a contrast enhanced MR examination, leading to an erroneous diagnosis of nerve sheath tumor. An attempt has been made to explain the likely mechanism of formation accounting for the imaging appearances, along with a review of the literature. PMID- 9730332 TI - Malignancy in pigmented villonodular synovitis. AB - Malignant pigmented villonodular synovitis is an extremely rare and controversial disease. We describe malignant change in pigmented villonodular synovitis of the ankle in a patient with an unusually long clinical history. Symptoms began at age 21, metastatic disease developed at age 85, and the patient died 1 year later. The histologic appearance of the malignant tumor differed from that in most reported cases, in that spindle-shaped cells predominated. Lymph node metastasis also developed, a feature uncommon to soft tissue sarcomas. PMID- 9730333 TI - Merkel cell carcinoma of the abdominal wall. AB - Merkel cell carcinoma is a rare highly malignant tumour. There have been previous descriptions of the CT appearances of this tumour, but to our knowledge this is the first MRI description. MRI may be a more sensitive method of initial evaluation of the local extension of the primary tumour. PMID- 9730334 TI - Post-traumatic bone cyst: a case of the floating fragment. AB - We report a case of an intraosseous foreign body leading to the formation of a bone cyst. The cyst was identified 20 years after the initial injury and was found to contain the foreign body. No infection was found. PMID- 9730335 TI - Upper extremity and rib stress fractures in a child. AB - Stress fractures in children are rare compared with the incidence in adults. This report describes an 11-year-old girl with stress fractures of the acromion, clavicle, and first rib on the left and contralateral fractures of the first and second ribs. It was eventually discovered that these fractures were caused by a nervous tic consisting of repetitive, vigorous shrugging and translation of the shoulders. PMID- 9730336 TI - Classic adamantinoma in a 3-year-old. AB - Classic adamantinoma of the long bones is a rare, low-grade malignant neoplasm arising most often in the tibia and usually in patients during the second to fifth decades. Although adamantinomas have been described in children, the histologic pattern in this age group is different from that seen in adults and resembles osteofibrous dysplasia. The usual pattern of adamantinoma in children has been termed "differentiated adamantinoma" and follows a benign course. We report a case of adamantinoma in the proximal tibia of a 3-year-old patient. The lesion had abundant epithelial component with formation of keratin pearls, a pattern that has been described only in classic adamantinoma occurring in adults. Since differentiated adamantinomas are essentially benign and classic adamantinomas are low-grade malignancies, the finding of a classic variant at this young age raised important therapeutic and prognostic issues. PMID- 9730337 TI - Anterior temporal white matter lesions in myotonic dystrophy with intellectual impairment: an MRI and neuropathological study. AB - We studied 12 patients with myotonic dystrophy using MRI and the Mini-mental state examination (MMSE), to see it specific MRI findings were associated with intellectual impairment. We also compared them with the neuropathological findings in an autopsy case of MD with intellectual impairment. Mild intellectual impairment was found in 8 of the 12 patients. On T2-weighted and proton density weighted images, high-intensity areas were seen in cerebral white matter in 10 of the 12 patients. In seven of these, anterior temporal white-matter lesions (ATWML) were found; all seven had mild intellectual impairment (MMSE 22-26), whereas none of the four patients with normal mentation had ATWML. In only one of the eight patients with intellectual impairment were white-matter lesions not found. Pathological findings were severe loss and disordered arrangement of myelin sheaths and axons in addition to heterotopic neurons within anterior temporal white matter. Bilateral ATWML might be a factor for intellectual impairment in MD. The retrospective pathological study raised the possibility that the ATWML are compatible with focal dysplasia of white matter. PMID- 9730338 TI - Improved imaging of the spinal cord in multiple sclerosis using three-dimensional fast spin echo. AB - We report assessment of a new three-dimensional fast spin echo (3D FSE) sequence in ten patients with clinically definite multiple sclerosis, comparing it with standard 2D FSE, and in ten normal controls. We saw 29 focal lesions on the 2D images and 53 on the 3D FSE images (P = 0.05); none were seen in controls. Lesion length was significantly smaller on the 3D FSE than on to the 2D FSE images (3D: 1.36; 2D 2.0; P = 0.03). This may relate in part to separation into several lesions on the 3D images of confluent abnormal signal seen on 2D and in part to detection of small lesions missed by the thicker 2D FSE slices (3 mm compared to 1.5 mm). The 3D FSE sequence looks promising in improving spinal cord imaging. PMID- 9730339 TI - MRI in acute disseminated encephalomyelitis following Semple antirabies vaccine. AB - I reviewed MRI findings in five patients with acute disseminated encephalomyelitis following vaccination with Semple antirabies vaccine. MRI in two patients with encephalitis features showed multiple white matter lesions in the cerebrum, cerebellar peduncles and brain stem. Two patients who had features of cord involvement showed signal alterations in the cord extending over a few segments. Asymptomatic lesions in the cerebrum were seen in two patients. In a patient with encephalomyelitis MRI 50 days later showed resolution of the lesions. The white matter lesions described were indistinguishable from those seen in acute disseminated encephalomyelitis following other infections. PMID- 9730340 TI - Morphological dissociation between visual pathways and cortex: MRI of visually deprived patients with congenital peripheral blindness. AB - MRI was used to study possible morphological changes in the visual system in 12 patients suffering from congenital blindness of peripheral (ocular) origin. While their optical pathways showed degeneration, hypoplasia or atrophy in 7 out of 12 cases, the occipital cortex appeared normal in all cases. This dissociation between afferent pathways and the cortex is contrary to the assumption that visually deprived cortex may undergo degeneration. The finding is congruent with evidence that the occipital cortex is used for other, nonvisual functions. PMID- 9730341 TI - Age-related changes in brain perfusion of normal subjects detected by 99mTc-HMPAO SPECT. AB - Previous functional imaging data generally show impairment in global cerebral blood flow (CBF) with age. Conflicting data, however, concerning age-related changes in regional CBF (rCBF) have been reported. We examined the relative rCBF in a sample of healthy subjects of various ages, to define and localize any age related CBF reduction. Twenty-seven healthy subjects (17 male, 10 female; mean age 49 +/- 15, range 26-71, median 47 years) were studied by 99mTc-HMPAO brain SPECT. The younger age group consisted of subjects below, the older group above 47 years of age, respectively. Analysis was performed by applying three preformed templates, each containing delineated regions of interest (ROIs), to three transaxial brain slices at approximately 4, 6, and 7 cm above the orbitomeatal line (OML). The average number of counts for each ROI was normalized to mean uptake of the cerebellum and of the whole brain slice. Globally, 99mTc-HMPAO uptake ratio normalized to cerebellum was significantly decreased in older subjects, affecting both hemispheres. A slight left-to-right asymmetry was observed in HMPAO uptake of the whole study group. It did not, however, change with age. Regionally, both cortical and subcortical structures of older subjects were involved: uptake ratio to cerebellum was significantly lower (after correction for multiple testing) in the left basal ganglia and in the left superior temporal, right frontal and bilateral occipital cortices at 4 cm above the OML. At 6 cm above the OML, reduced uptake ratios were identified in the left frontal and bilateral parietal areas. At 7 cm, reduced uptake was detected in the right frontal and left occipital cortices. Most of these differences were reduced when uptake was normalized to whole slice, whereas an increase in uptake ratios was observed in the cingulate cortex of the elderly. An inverse correlation between age and HMPAO uptake ratios normalized to cerebellum was observed in a number of brain regions. These findings suggest that advancing age has a differential effect on cerebral perfusion reflected in brain 99mTc-HMPAO uptake. PMID- 9730342 TI - Delayed cerebral radionecrosis with a high uptake of 11C-methionine on positron emission tomography and 201Tl-chloride on single-photon emission computed tomography. AB - A 47-year-old woman, who 2.5 years previously had undergone resection of a malignant astrocytoma of the left temporal lobe followed by radiotherapy, was found to have a mass in the left frontal lobe. This showed high uptake of thallium-201 (201Tl) on single-photon emission computed tomography and 11C methionine on positron-emission tomography, suggesting recurrent tumour. Histological examination of the resected lesion, however, revealed it to be radionecrosis. This case thus illustrates a diagnostic pitfall in the use of these investigations for distinguishing radionecrosis from recurrent malignant glioma. PMID- 9730343 TI - Radiation-induced optic neuropathy 4 years after radiation: report of a case followed up with MRI. AB - We report a case of radiation-induced optic neuropathy in a 32-year-old man with Cushing's disease and a recurrent tumour of the left cavernous sinus. The patient experienced rapid, painless loss of vision 4 years after treatment without recurrence of tumour or other visual disorder. MRI showed enlargement and contrast enhancement of the optic chiasm. A year later the patient was almost blind and MRI showed atrophy and persistent contrast enhancement of the chiasm. PMID- 9730344 TI - Intracranial arterial dissection. AB - We review the angiographic and CT findings, precipitating factors and clinical features in nine patients with ten intracranial arterial dissections. The internal carotid artery was involved in five cases, the vertebral artery in four and the posterior inferior cerebellar artery in one. Angiography revealed irregular stenosis in four cases, irregular stenosis and a pseudoaneurysm in two, irregular stenosis and irregular dilatation in one, arterial occlusion in two and a pseudoaneurysm in one. CT demonstrated an infarct in four cases, a dense middle cerebral artery in two and subarachnoid haemorrhage in one. A possible precipitating factor was identified in five cases. Six patients recovered well, while three had persisting neurological deficits. PMID- 9730345 TI - Diffusion-weighted echo-planar MRI of lacunar infarcts. AB - We studied 35 patients with lacunar infarcts, using diffusion-weighted echo planar imaging (DW-EPI) at 1.5 T. The relative apparent diffusion coefficient ratio (ADCR) of each lesion was calculated and lesion conspicuity on DW-EPI was compared to that on images acquired with fast fluid-attenuated inversion recovery and T2-weighted fast spin-echo sequences. Acute small infarcts (within 3 days) were identified with DW-EPI as an area of decreased ADCR (range 0.33-0.87; mean 0.67) and high signal, subacute small infarcts (4-30 days) as a high-signal or isointense areas of decreased or nearly normal ADCR (0.54-0.98; 0.73), and chronic small infarcts (> 30 days) as low- or high-signal areas of nearly normal or increased ADCR (0.97-1.92; 1.32). In three patients, small infarcts of the brain stem in the hyperacute phase (within 6 h) were seen only with DW-EPI. In five patients, fresh small infarcts adjacent to multiple old infarcts could be distinguished only with DW-EPI. PMID- 9730346 TI - Epidermoid cyst of the skull with nonpulsatile tinnitus. AB - We report an intradiploic epidermoid cyst of the skull responsible for transverse sinus compression and presenting with nonpulsatile tinnitus. Plain films and CT both demonstrated the tumour. Cerebral angiography showed best the degree of narrowing of the right transverse sinus, accompanied with turbulent flow probably leading to tinnitus. MRI demonstrated accurately both the tumour and the dural sinus compression. The tumour was totally removed, cranioplasty was performed, and the patient was discharged free of symptoms. PMID- 9730347 TI - Spinal cord abscess in a heroin addict: case report. AB - Spinal cord abscesses are extremely rare, even in intravenous drug abusers. They usually have a poor prognosis unless diagnosed and treated promptly. MRI is the best imaging modality for diagnosis and follow-up. We report a 42-year-old man, an active intravenous drug user, HIV negative, who developed subacute tetraplegia from an intramedullary abscess caused by Staphylococcus aureus. Immediate decompressive surgery and antibiotic treatment led to progressive recovery. PMID- 9730348 TI - Arteriovenous fistula as a complication of lumbar disc surgery: case report. AB - Arteriovenous fistula (AVF) is a rare, late complication of lumbar disc surgery. It is often not suspected and the symptoms are diagnosed as heart failure or deep venous thrombosis. We report a case in which the patient developed leg swelling and high-output congestive heart failure due to a left ilioiliac AVF after lumbar laminectomy. PMID- 9730349 TI - CT of the temporal bone in the CHARGE association. AB - We reviewed the CT examinations of the temporal bone, performed with 1-mm-thick contiguous sections, of seven patients with the CHARGE association. We found abnormalities of the incus and stapes, with ossicular chain fixation, absence of the stapedius muscle and oval window, hypoplasia or dysplasia of the vestibule and absence of the semicircular canals in all ears. The pyramidal eminence and tympanic sinus were absent and there were anomalies of the cochlea in 13 of 14 ears. Absence of the semicircular canals is the most specific change in patients with the CHARGE association. PMID- 9730350 TI - Enhanced catheter propagation with hypercapnia during superselective cerebral catherisation. AB - During arterial catherisation of a cerebral arteriovenous malformation it may be difficult or impossible to access the nidus of the malformation through its small, tortuous feeding vessels due to microcatheter impaction. Carbon dioxide, a most potent cerebral vasodilator, was temporarily added to the inspired gases of two anaesthetised patients undergoing superselective embolisation of an arteriovenous malformation, when the microcatheter had been impacted for a considerable time. Successful propagation of the microcatheter into the malformation was achieved in both patients after a relatively short period of hypercapnia. PMID- 9730351 TI - The need for repeat angiography in subarachnoid haemorrhage. PMID- 9730352 TI - Inferior sagittal sinus thrombosis. PMID- 9730353 TI - Diversity, identity, and adolescent health. PMID- 9730354 TI - Driver education. PMID- 9730355 TI - Structural equation socialization model of substance use among Mexican-American and white non-Hispanic school dropouts. AB - PURPOSE: To test a socialization model of polydrug use among Mexican-American and white non-Hispanic school dropouts. METHODS: A sample of 910 Mexican-American and white non-Hispanic school dropouts were surveyed regarding their use of alcohol, marijuana, and other drugs, and socialization characteristics that have previously been shown to be predictive of adolescent substance use. A structural equation model based on peer cluster theory was evaluated for goodness of fit and for differences in model characteristics by ethnicity and gender. RESULTS: Results partially confirmed peer cluster theory among school dropouts in that association with drug-using peers was the most powerful direct predictor of substance use. The effects of a number of other socializing influences were indirect, mediated through association with drug-using peers. Some differences were present between Mexican-American and white non-Hispanic subgroups. CONCLUSIONS: Results were similar to those obtained from previous tests of this model among youth who remain in school, suggesting that social influences on drug use are similar across students and school dropouts. Association with drug-using peers dominates the prediction of substance use among school dropouts. However, family communication of drug use sanctions helps to both limit substance use and strengthen family bonds. Prior school adjustment is likely to be an important protective factor in limiting substance use among Mexican-American dropouts. PMID- 9730357 TI - Whither "culture" in adolescent health research? PMID- 9730356 TI - Skills training for pregnancy and AIDS prevention in Anglo and Latino youth. AB - PURPOSE: This study tested social skills training (SST), didactic training (DT), and no training (NT) on adolescents' social skills for resisting peer pressure to engage in acquired immunodeficiency syndrome (AIDS) and pregnancy risk behavior. METHODS: A total of 307 Latino and Anglo youth ages 13-18 years were assigned at random to receive 18 h of SST, 18 h of DT, or NT. RESULTS: Significantly (p < 0.05) greater increases in assertiveness followed SST compared to DT or NT for three trained skills: condom negotiation, asking a friend about their sex/drug history, and discussing a friend's risk of AIDS. Untrained negotiation skills (e.g., purchasing a condom) did not increase significantly. SST did not result in increased assertiveness for refusal skills. DT increased knowledge of AIDS significantly more than SST; both DT and SST increased knowledge significantly more than NT. CONCLUSIONS: Social skills training can increase assertiveness for certain negotiation skills that may decrease risk of AIDS for Latino, Anglo, and male and female adolescents. Both DT and SST can increase knowledge of AIDS prevention. Differences between experimental groups were supported by differences between trained and untrained skills within the SST condition, adding to discriminant validity. PMID- 9730358 TI - Association between violent behaviors and substance use among Mexican-American and non-Hispanic white high school students. AB - PURPOSE: To determine the prevalence of violent behaviors among Mexican-American and non-Hispanic white high school students and to explore the associations between violent behaviors and alcohol and illicit drug use. METHODS: The Youth Risk Behavior Survey was administered to 1786 high school students in a biethnic community in Southeast Texas; 65% were Mexican-American, 26% were non-Hispanic white, and 9% were of another ethnicity. RESULTS: There were no significant ethnic differences in prevalence of drinking alcohol, illicit drug use, fighting, carrying a weapon, or planning or attempting suicide. After adjustment for age, carrying a weapon and fighting were significantly associated with alcohol and illicit drug use, with few exceptions, among the four gender- and ethnic-specific subgroups. However, the relationship between suicide (plans and attempts) and substance use among the four subgroups was less consistent and of much lower magnitude than for carrying a weapon and fighting. CONCLUSIONS: A substantial percentage of adolescents engage in violent behaviors, and fighting and weapon carrying are associated with substance use among both gender and ethnic groups. A systematic and integrated approach to changing the environment and norms of communities is needed to affect change and reduce the morbidity and mortality associated with violent behaviors. PMID- 9730359 TI - Ethnic and gender differences in smoking prevalence among a longitudinal sample of inner-city adolescents. AB - PURPOSE: To determine if ethnic and gender differences in smoking (lifetime prevalence and 30-day prevalence) exist among a cohort of Asian, black, Hispanic, and white inner-city adolescents during the 3-year middle school period. METHODS: Students in 22 urban schools completed self-report questionnaires and provided carbon monoxide breath samples at three annual assessments. Chi-square analyses were conducted to test for associations between ethnic group (Asian, black, Hispanic, and white) and smoking and to test for gender differences in smoking within each ethnic group. Additional analyses examined differences in smoking between two Hispanic subgroups (Puerto Rican and Dominican). RESULTS: Ethnicity was associated with lifetime smoking prevalence at all three assessment points but was only associated with 30-day smoking prevalence at the 2-year follow-up. However, there were no differences in smoking between Puerto Rican and Dominican youth. Black girls reported higher lifetime smoking prevalence than black boys at all three assessments. At the 2-year follow-up, Asian boys reported higher lifetime smoking prevalence than Asian girls; Hispanic girls reported higher 30 day prevalence than Hispanic boys. CONCLUSIONS: White and Hispanic adolescents were at higher risk for smoking relative to Asian and black adolescents. With the exception of white youth, gender differences were found within each ethnic group. PMID- 9730360 TI - Ethnic differences in correlates of adolescent cigarette smoking. AB - PURPOSE: To examine the correlates of cigarette smoking among African-American, Hispanic, and white adolescents in a cross-sectional national sample. METHODS: A total of 1795 mother-child dyads from the 1992 National Longitudinal Survey of Youth were selected for analyses. Measures of adolescents cigarette smoking and family, individual, peer, and sociodemographic risk factors were analyzed. RESULTS: White youths reported the highest rates of lifetime, current, and persistent smoking, and initiated smoking at a significantly earlier age than African-Americans and Hispanics. Except for maternal cigarette smoking and substance use, African-Americans and Hispanics experienced a disproportionately larger number of purported risk factors than whites. Multivariate analyses revealed common and ethnic-specific correlates of adolescent lifetime and current smoking, with many more significant associations among whites than minorities. Common correlates included youth's age across all three ethnic groups, problem behaviors and delinquency among whites and African-Americans, and perceived peer pressure to smoke among whites and Hispanics. Ethnic-specific correlates included maternal smoking, maternal cocaine use, low maternal religiosity, and negative scholastic attitudes, which increased smoking for whites; and positive parenting, which reduced smoking for African-Americans. CONCLUSIONS: The lack of effects of maternal smoking and perceived peer pressure to smoke on African-American adolescents compared with whites suggests that role modeling and interpersonal influence may be more important determinants of smoking for white than African American adolescents. The differential impact of family and peer factors on the smoking of adolescents of different ethnicity warrants further investigation. PMID- 9730361 TI - Gender differences in health and risk behaviors among bisexual and homosexual adolescents. AB - OBJECTIVE: This study explored gender differences in the health and risk behaviors of 394 self-identified bisexual and homosexual adolescents who participated in an anonymous, school-based survey. METHODS: Respondents included 182 girls and 212 boys; girls were significantly younger than boys (p < 0.001), so respondents were further grouped as younger (< or =14 years) and older (> or =15 years) for analysis. Chi-square was used to test for gender differences in health perceptions and risk behaviors. Items included general health perceptions and health care access, body image and disordered eating behaviors, sexual behaviors, alcohol use, and emotional health measures including mood, life satisfaction, and suicidal ideation and attempts. RESULTS: Both younger and older girls were significantly more likely than their male age mates to report a history of sexual abuse, dissatisfaction with weight, a negative body image, more frequent dieting, and an earlier age at onset of sexual intercourse. Both younger and older boys were significantly more likely than girls to have a positive body image, to rate themselves as healthier than peers, to report no regular source of health care, to be sexually experienced, and to drink alcohol more often and in greater quantity; a significantly greater proportion of older boys than older girls reported alcohol use before school (19.0% vs. 3.9%; p < 0.05). No significant gender differences were found for measures of emotional health, including suicidal ideation and attempts; however, nearly 1 of 3 older boys and girls reported at least one suicide attempt. CONCLUSIONS: Gender is a substantive source of variation in health and risk behaviors among bisexual and homosexual adolescents. Health care providers should incorporate gender-specific approaches to health promotion and risk reduction with young people who self-identify as gay, lesbian, or bisexual. PMID- 9730362 TI - Partitioning of oxygen and carbon monoxide in the three human embryonic hemoglobins. AB - The ratio of oxygen to carbon monoxide binding to the three fully saturated human embryonic hemoglobins has been determined. The embryonic hemoglobins exhibit significantly lower values of carbon monoxide binding than any previously reported mammalian fetal or adult hemoglobins. The possible protective role of this with regards to carbon monoxide intoxication is discussed. These data are combined with previous parameters yielding a significant correlation between oxygen affinity and carbon monoxide binding. A possible molecular origin of this correlation is discussed. PMID- 9730363 TI - A novel epitope (pentapeptide) in the human hemoglobin beta chain. AB - We have cloned 16 monoclonal antibodies by immunizing mice with human hemoglobin for the purpose of analyzing epitopes in human hemoglobin. By using one, SU-115, which is specific for the beta chain, an epitope in the beta chain reacting with this monoclonal antibody was investigated, and a pentapeptide was identified as a novel epitope. After digestion of the beta chain by lysylendopeptidase, the antigenicity of degradation products was examined. An antigenic fragment against SU-115 was found to be a peptide corresponding to residues 96-120 of the beta chain by amino acid analysis of its N-terminal region. Several peptides involved in the region of beta96-120 were synthesized to be examined for their reactivity to SU-115 using dot-blot analysis and a resonant mirror detection method, and a pentapeptide (N108VLVC) was determined as a major sequence of an epitope. By injecting this pentapeptide into a mouse, an antibody reacting human hemoglobin (alpha2beta2) in the same order of strength as SU-115, was obtained. The pentapeptide described in this paper seems to be the minimum size as a major sequence of the actual epitope in human hemoglobin so far reported, and in the primary structure of this region (108-112) there is a difference of three amino acids between human and mouse hemoglobin. PMID- 9730364 TI - Molecular characterization of beta-thalassemia in Pakistan. AB - Beta-thalassemia is one of the most common inherited hemoglobin disorders in Pakistan. The carrier frequency is estimated to be 5.4%. To determine the spectrum of beta-globin gene defects causing beta-thalassemia, we have analyzed a representative sample of 602 alleles from six ethnic groups in Pakistan; 99.2% alleles were characterized, while 0.8% remained unidentified. The spectrum of mutations is heterogeneous and we have found 19 different mutations in all ethnic groups. The four most common mutations, IVS-I-5 (G-->C) (37.7%), codons 8/9 (+G) (21.1%), the 619 bp deletion (12.4%), and IVS-I-1 (G-->T) (9.5%), account for 80.7% of the alleles. There are differences between the ethnic groups and also between provinces. In the four provinces of Pakistan, the IVS-I-5 (G-->C) mutation is more prevalent in Sindh and Balochistan, bordering India in the south and Iran in the southwest, while the codons 8/9 (+G) mutation is more common in the Punjab and the North West Frontier Province, bordering India in the northeast and Afghanistan, respectively. The 619 bp deletion is high (46%) in Gujratis and Memons residing in the Province of Sindh, neighboring the Indian Gujrat. PMID- 9730365 TI - Hb Oita [alpha45(CE3)His-->Pro]: a new silent hemoglobin variant. AB - We describe a new alpha chain variant accidentally found in a 49-year-old female living in Usa City, Oita Prefecture, Japan. An abnormally low Hb A1c value of 2.5% (normal range: 4.8-6.3%) was found while she was treated with glucocorticoid for Fisher syndrome. The patient was also diagnosed as having an iron deficiency anemia, but otherwise showed a normal hemogram. An abnormal hemoglobin was not detectable by isoelectrofocusing and high performance liquid chromatographic methods, but appeared as a fast-moving alpha chain abnormality by urea carboxymethyl cellulose column chromatography of the globin, from which the content of the abnormal hemoglobin was estimated at approximately 20%. The instability test of the hemolysate was normal. Structural studies demonstrated that the abnormal hemoglobin had an amino acid substitution of His-->Pro at alpha45. It is a new variant and was named Hb Oita or alpha45(CE3) His-->Pro. Additionally, sequence analysis showed a nucleotide change from A-->C at the second base in the 45th codon of the alpha2 gene, CAC(His)-->CCC (Pro). The beta/alpha ratio was 0.51 (normal range: 0.9-1.2). Her mother and a son did not have the abnormal hemoglobin variant; her father was deceased and no sample was available to verify the inheritance of the variant in this kindred. PMID- 9730367 TI - Detection of alpha2- and alpha1-globin gene variants by a modified cycle sequencing method. PMID- 9730366 TI - A new hemoglobin variant found during Hb A1c measurement: Hb Hokusetsu [beta52(D3)Asp-->Gly]. AB - A new beta chain variant was accidentally found through the assay of Hb A1c in a diabetic patient. The variant was detected by polyacrylamide gel isoelectrofocusing and electrospray ionization mass spectrometry. For sequence determination, globin was cleaved with combination of trypsin and lysyl endopeptidase and analyzed by high performance liquid chromatography connected to electrospray ionization mass spectrometry. An abnormal betaT-5 peptide was found by reconstructed selected ion monitoring. The collision-induced dissociation spectrum of an ion derived from the abnormal betaT-5 peptide revealed a new substitution, [beta52(D3)Asp-->Gly], named Hb Hokusetsu. The sequence was confirmed with an automatic sequencer using peptides isolated by reversed phase high performance liquid chromatography. Amplification of the beta-globin exon 2 and nucleotide sequencing revealed a GAT-->GGT mutation in codon 52 corresponding to an Asp-->Gly replacement. Electrospray ionization mass spectrometry analysis of the hemolysate showed a reasonable value of 10.4% for glycated globin. The variant migrated as Hb S on isoelectrofocusing. Hematological analysis revealed normal parameters. The patient's hemolysate showed normal stability in the isopropanol test. Oxygen equilibrium studies on the patient's red blood cells and hemolysate showed no significant change in oxygen affinity or cooperativity. PMID- 9730368 TI - Another example of the beta-thalassemia mutation, IVS-I (-2) or codon 30 (A-->G), found in a Chinese family. PMID- 9730370 TI - A novel polymorphism 3' to the beta-globin gene. PMID- 9730369 TI - Severe Hb S-beta+ thalassemia caused by IVS-I splice site mutations. PMID- 9730371 TI - A new beta chain variant, Hb Vienna or beta77(EF1)His-->Gln. PMID- 9730372 TI - Hb Brockton [beta138(H16)Ala-->Pro] observed in a Chinese boy. PMID- 9730373 TI - Molecular mechanisms of hepatitis B and C viruses related to liver carcinogenesis. PMID- 9730374 TI - Pathological aspects of hepatocellular carcinoma: a critical review of prognostic factors. AB - With surgical advances the number of hepatectomy specimen has increased, leading in turn to growing information on the pathologic characteristics of hepatocellular carcinomas. Large series have delineated prognostic pathological indicators of recurrence as well as survival. The understanding by histopathologists as well as surgeons of the importance of the detailed analysis is crucial since it may dictate the therapeutic strategy. We review herein the pathological factors that are currently recorded at specialized centers and analyze their value as prognostic indicators for patients with HCC. PMID- 9730375 TI - Pathology of early hepatocellular carcinoma: progression from early to advanced. AB - Small, early stage hepatocellular carcinoma (HCC) can be divided into 2 types; small nodular HCC with distinct margins and small HCC with indistinct margins. The latter consists of well-differentiated cancerous tissue with replacing growth at the boundary and with many portal tracts retained in the tumor. When these tumors reach approximately 1.5-2.0 cm in diameter, moderately or poorly differentiated cancer tissues develop within the well-differentiated cancer tissue and well-differentiated cancer tissues are replaced by less differentiated cancer tissues. This dedifferentiation seems to be closely related to tumor proliferation. When less differentiated cancer tissues within the well differentiated cancer nodules proliferate in an expansive fashion, a "nodule in nodule" appearance is frequently identified. On the other hand, small nodular HCCs with distinct margins are well-defined, with more than half of them encapsulated by a thin fibrous capsule, and about 60% moderately differentiated. Tumor invasion into the portal vein and intrahepatic metastasis are found in 27% and 10%, respectively. Thus, although the size of the tumor may be small, some distinctly nodular small HCCs can already be interpreted as advanced cancers. PMID- 9730376 TI - Progression of hepatitis B and C to hepatocellular carcinoma in Western countries. AB - Hepatitis B virus (HBV) and hepatitis C virus (HCV) are major causes of chronic liver disease and hepatocellular carcinoma (HCC) worldwide. In Western countries most HCC associated with HBV or HCV infection occurs in the cirrhotic liver. In a longitudinal studies of series of a Caucasian patients with compensated cirrhosis the incidence of HCC was approximately 2 and 2.5 per 100 person-years for HBV related and HCV-related cirrhosis, respectively. The host factors of age, age at infection and male sex as well as parameters reflecting the severity of liver disease, such as bilirubin and platelets, appear to be significant predictors of liver cancer in patients with cirrhosis. It is still controversial whether HCV genotype 1b has a specific oncogenic potential. Studies from Europe have shown a low risk for HCC in patients with compensated cirrhosis type B who achieve a sustained virological and disease remission. Overall these data strengthen the epidemiological evidence of the association among HBV infection, cirrhosis and HCC in the West, indicating a similar risk for liver cancer in Caucasian patients with cirrhosis type B or C and suggest that tumors occur mainly in patients with long-standing disease. Additional factors that may promote liver cancer include concurrent HBV and HCV infection (approximately 5 fold-increase in risk) or concomitant heavy alcohol intake (synergism). PMID- 9730377 TI - Natural course of small hepatocellular carcinoma with underlying cirrhosis. A study of 30 patients. AB - To anticipate the prognosis and choose therapy for patients with relatively early stage HCCs, we elucidate the natural course of such patients. Thirty cirrhotic patients with small hepatocellular carcinoma (HCC) < 3 cm in size and not receiving anti-cancer treatment were followed by sonography for periods of 6-48 mots. The growth speed varied considerably from case to case, with an average of 6.5+/-5.7 months of doubling time (DT). Growth speed of small HCC is significantly related to histological differentiation of tumors: a slow-growing HCC tends to be well-differentiated and a rapid-growing HCC moderately- or poorly differentiated. Furthermore, there was a tendency to grow without changing from nodular type in slow-growing HCCs and to multiple nodular or massive types in intermediate- and rapid-growing HCCs. The survival rates of these untreated HCC patients showed a 1-yr survival of 90.7%, a 2-yr survival of 55.0%, and a 3-yr survival of 12.8%. Factors significantly influencing survival of these patients were serum total bilirubin level and DT. This study will provide invaluable data for diagnosing and treating for small HCCs. PMID- 9730378 TI - The natural history of hepatocellular carcinoma in Western countries. AB - Hepatocellular carcinoma (HCC) often has a long-lasting subclinical incubation period during which it may grow as a solitary mass. This does not mean that, in some patients, the tumor may be found as more than one nodule when it is first detected. The so-called expanding, encapsulated type of tumor seems to prevail among Oriental, Italian and French patients, rather than the more aggressive, spreading form of tumor. However, many patients with apparently single encapsulated HCC nodes may have ancillary tumors detectable only with highly sensitive imaging techniques or during surgery. There are important variations in the growth rate of this tumor, even in a single patient with multiple nodes. These differences in the growth patterns of HCC have made a clear understanding of the natural history of this tumor and prediction of patient survival difficulty. Thus, survival times are better correlated with the severity of liver impairment than with the number or size of tumors. Hepatic failure and gastrointestinal bleeding are the leading causes of death in both Oriental and Caucasian patients, accordingly. There are still many aspects of the natural history of HCC to be investigated, including whether or not all HCCs originate from one clone of transformed liver cells or more, as well as identifying which tumor and/or patient characteristics may predict individual tumor growth rates. PMID- 9730379 TI - Morphological diagnosis of hepatocellular carcinoma: special emphasis on intranodular hemodynamic imaging. PMID- 9730380 TI - Hepatocellular carcinoma: radiological findings. PMID- 9730381 TI - Transcatheter arterial chemoembolization of hepatocellular carcinoma. PMID- 9730382 TI - Intra-arterial chemoembolization in patients with hepatocellular carcinoma. AB - Hepatocellular carcinoma occurs almost exclusively in patients with cirrhosis, at least in the West. In most of these patients, potential curative treatments, such as resection or percutaneous alcohol injection, are usually contra-indicated. Transarterial chemoembolization may induce tumor necrosis. In order to avoid massive necrosis of the non tumoral liver, two major contra-indications have been identified: inadequate portal flow and liver failure. The influence of chemoembolization on survival was thought to be high on the basis of non randomized trials. However, no beneficial effects on survival were observed in three randomized trials. In these trials, the beneficial effect on tumor necrosis was counterbalanced by frequent deleterious effects on liver function. Moreover, progressive liver atrophy may follow repeated procedures. As there is no alternative treatment for most of these patients and chemoembolization can still be beneficial in selected cases, efforts have been made to improve patient selection and method to improve the results. Good liver function, a normal portal flow, and a well limited hypervascularized tumor are necessary conditions for treatment, which may even be curative when used in association with percutaneous alcohol injection. Moreover, arterial embolization can be performed without chemotherapy, and the procedure should not be repeated in the short term. PMID- 9730383 TI - Percutaneous Ethanol Injection in the treatment of hepatocellular carcinoma in cirrhosis. PMID- 9730384 TI - Comparison of percutaneous acetic acid injection and percutaneous ethanol injection for small hepatocellular carcinoma. AB - BACKGROUND/AIMS: To assess whether ultrasound (US)-guided percutaneous acetic acid injection is more effective than percutaneous ethanol injection in the treatment of small hepatocellular carcinoma (HCC). METHODOLOGY: sixty patients with 1 to 4 HCC smaller than 3 cm entered a randomized controlled trial from August 1993 to September 1995. Thirty one and 29 patients were treated by percutaneous acetic acid injection using 50% acetic acid and percutaneous ethanol injection using absolute ethanol, respectively. There were no significant differences in clinical characteristics and biochemical data between the two groups. RESULTS: All original tumors were treated successfully by the chosen therapy. However, local recurrence occurred in 8% of the 38 tumors treated with percutaneous acetic acid injection and 37% of the 35 tumors treated with percutaneous ethanol injection P>0.001). The 1- and 2-year survival rates were 100% and 92% with percutaneous acetic acid injection and 83% and 63% with percutaneous ethanol injection (p=0.0017). Multivariate analysis of prognostic factors revealed that treatment was an independent predictor of survival. CONCLUSION: percutaneous acetic acid injection is more effective than percutaneous ethanol injection. PMID- 9730385 TI - Chemotherapy in hepatocellular carcinoma. AB - Systemic chemotherapy for hepatocellular carcinoma (HCC) has been of limited value in clinical practice, because only a small portion of patients obtain significant effects, and because the toxicity of chemotherapy often outweighs the benefits. Therefore, at present, only HCC patients with no indication for standard treatments (surgical resection, percutaneous ethanol injection and transcatheter arterial embolization) undergo chemotherapy. Due to the causes of death, tumor response and survival rate, only patients with metastatic sites other than in the bone, patients with no tumor thrombus in the main portal trunk and with good hepatic reserve are good candidates for systemic chemotherapy. To improve the prognosis in HCC patients, effective chemotherapy must be developed. The recommended strategy is to include all patients with advanced HCC in well designed phase II trials with novel anti-cancer agents or regimens, when patients can tolerate treatment. Furthermore, prospective randomized trials comparing systemic chemotherapy with other palliative therapies or the best supportive care for HCC, are necessary before a clinically available chemotherapeutic regimen can be recommended for HCC. PMID- 9730386 TI - Chemotherapy and medical treatment of hepatocellular carcinoma (HCC). PMID- 9730387 TI - Hepatic resection for hepatocellular carcinoma -- Japanese experience. AB - In the past 20 years, thanks to the early detection of hepatocellular carcinomas (HCCs), good perioperative care, the evaluation of functional liver reserve, preoperative portal embolization and the improvement in surgical techniques such as intraoperative ultrasonography, the surgical resection of HCC has become very safe. We have performed 367 hepatectomies on 352 patients since 1990 with a surgical mortality, hospital mortality, blood transfusion rate and 5-year survival rate of 0.27, 0.82, and less than 10 and 47.4%, respectively. Our standard method for selecting surgical procedures and perioperative care resulting in low blood transfusion rates and almost no mortality are described. Since 1990, ethanol injection for HCC ablation has been extensively used in Europe and Japan, but results are poorer than with surgical intervention. Therefore, in patients with small HCCs and good liver function, the first choice treatment should not be ethanol injection, but surgical resection. PMID- 9730388 TI - Aggressive management of recurrence following surgical resection of hepatocellular carcinoma. AB - BACKGROUND: Liver resection of hepatocellular carcinoma (HCC) is associated with a high incidence of recurrence, that has a poor prognosis. AIM: Assess the rationale for and result of an aggressive treatment of recurrence following resection of HCC. METHODOLOGY: Retrospective analysis of 132 patients with recurrent HCC with special reference to the topography and time of onset of recurrence as well as outcome following treatment of these recurrences. Case control analysis of the efficacy of repeat hepatectomy and its influence on the long term prognosis of patients with recurrent tumor. RESULTS: Sixty seven percent of the recurrences were exclusively intrahepatic and half of these were limited in size and number. The 5-year survival rate following treatment of limited intrahepatic recurrence by repeat hepatectomy, arterial chemoembolization or percutaneous ethanol injection was 30%. Repeat hepatectomies improved the long term outcome of patients with recurrent HCC. CONCLUSION: An aggressive approach to tumor recurrence is currently the best way to improve the long term outcome following resection of HCC. PMID- 9730390 TI - Problems of hepatectomy in cirrhosis. PMID- 9730389 TI - Surgical treatment of hepatocellular carcinoma on cirrhosis: a Western experience. AB - Recent improvements on the therapeutical management of hepatocellular carcinoma (HCC) on cirrhosis have led to further evaluate the role of surgery for this disease. In a 15-year period we have evaluated 532 cirrhotics with HCC on cirrhosis. Contraindications for surgery were founded in 170 (31.9%); 37 of them received a transarterial chemoembolization and 2 a percutaneous ethanol injection. Laparotomy was performed in 315 (59.2%) cases, but in 77 surgical treatment was contraindicated due to unexpected intraoperative findings. A liver resection was performed in 238 (44.7%) patients, representing the 26.1% of all liver resections performed at our Department. Seventy-eight (32.8%) were subsegmentectomies, 143 (60.1%) segmentectomies (including 1 to 3 anatomical segments) and 17 major hepatectomies. Overall 30-day mortality was 4.6%: 9.3% during years 83-91 and 0.8% during following years (P<0.005). Five-year actuarial survival rate was 41.3%. The remaining 47 (8.8%) patients were placed on the waiting list for orthotopic liver transplantation (OLT) and 41 already operated on. Operative mortality was 6.2% and 5-year actuarial survival rate 58.1%. The persistent shortage of organ donor represents the major factor limiting the application of liver transplantation for a larger number of patients carrying HCC on cirrhosis. Liver resection remains the option to be considered for all the patients with such a disease, even if in a large proportion of cases this procedure offers only a limited survival. PMID- 9730391 TI - Prevention of the occurrence of hepatocellular carcinoma in patients with cirrhosis. PMID- 9730392 TI - Caring for the caregiving surgeon. PMID- 9730393 TI - A surgical option for familial chylomicronemia associated with insulin-resistant diabetes mellitus. AB - BACKGROUND: The goal of the present work is to present an effective surgical approach for the treatment of a medically-resistant form of hyperlipidemia. METHODS: Two siblings with familial lipoprotein-lipase deficiency and subsequent hyperchylomicronemia, widespread skin xanthomas and severe insulin-resistant diabetes mellitus came to our observation after several unsuccessful attempts at medical treatment. In order to lower plasma lipids through lipid malabsorption, a modified bilio-pancreatic diversion operation was employed. The rationale in deciding to use this surgical approach was based also on the likely hypothesis that diabetes, in these subjects, was secondary to high circulating and tissue levels of lipids. Insulin sensitivity in the two treated subjects, as well as in 24 healthy volunteers constituting the control group, was assessed by euglycemic hyperinsulinemic clamp and indirect calorimetry, obtaining total end-clamp glucose uptake (M) and end-clamp glucose oxidation (ECGO) rates. RESULTS: Within 3 weeks of surgery, plasma triglycerides and cholesterol levels had decreased from 4500 and 500 mg/dl (with dietary restrictions) to lower than 450 and 150 mg/dl (on a free, lipid-rich diet) respectively. Fasting plasma glucose levels had decreased from above 300 (under daily repeated subcutaneous injections of insulin) to 80-100 mg/dl (without administration of insulin or oral hypoglycemic agents). Body weight and fat free mass were maintained in both subjects after surgery. In both patients, before surgery M and ECGO were significantly lower than in normal subjects, while after surgery they were not significantly different from normal subjects, confirming the positive metabolic effect of the operation. CONCLUSION: The surgical option used in these patients may represent an interesting and effective new possibility for treatment of those severe cases of hyperlipemia leading otherwise to metabolic complications and a low quality of life. PMID- 9730394 TI - Esophageal anatomy and function in laparoscopic gastric restrictive bariatric surgery: implications for patient selection. AB - BACKGROUND: The purpose of this study was to assess factors of clinical importance in morbidly obese patients having a laparoscopically adjustable gastric band (LAP-BAND) implanted in order to achieve weight loss. METHODS: Preoperative evaluation of hiatus hernia and esophageal (dys)motility were compared with the need for reoperation. Results are presented for the first 50 consecutive patients entered. RESULTS: Nine of the first 50 patients required reoperation (18%). Five (10%) were for LAP-BAND slippage on the stomach. Of these five, reoperation was required in four of 12 (33%) with hiatus hernia (P = 0.0093); three of nine (33%) with a motility disorder (P = 0.025); and three of six (50%) with both hiatus hernia and a motility disorder (P = 0.0076). CONCLUSIONS: We identify two factors, hiatus hernia and esophageal dysmotility, which are associated, both independently as well as in combination, with reoperation for LAP-BAND slippage. Both patients and their physicians should consider these data when considering the LAP-BAND as possible therapy for morbid obesity. PMID- 9730396 TI - Periprosthesis sepsis: an undesirable complication of silicone gastric banding. AB - BACKGROUND: Gastric banding is a very satisfactory procedure for the treatment of morbid obesity. The significant incidence of skin suppuration in these patients makes the laparoscopic approach a suitable technique. Regardless of this, in some cases, suppuration can still rarely result. METHODS AND RESULTS: In four patients the authors observed diffusion of suppuration in both directions along the catheter which connects the port to the band, necessitating band removal and thus invalidating the procedure. CONCLUSIONS: Suppuration of port location is an undesirable complication that must be avoided because it may contaminate the entire device system. This complication must be carefully evaluated for a correct diagnosis and an eventual removal of the band. PMID- 9730395 TI - Adjustable silicone gastric banding: complications in a personal series. AB - BACKGROUND: Morbid obesity is a very severe pathology, deriving partly from a psychological disturbance of nutritional behavior. Besides a behavioral therapeutic approach, surgery appears to be necessary to resolve associated diseases by causing a satisfactory weight loss. Adjustable gastric banding is a less-invasive, potentially reversible procedure that guarantees an optimal quality of life. METHODS: The authors have performed Kuzmak's gastric banding since 1992, with the lap-band approach since 1995; 183 patients were submitted to surgery, and 68 of these were operated by the laparoscopic approach. Average body mass index was 45.5 kg/m2. The complications were always under control and have decreased since the introduction of the recent lap-band. RESULTS: Gastric banding is still a very young procedure and it is difficult to state definitive results yet. Preliminary results, according to our experience are satisfactory in terms of weight loss, without metabolic changes and without mortality. CONCLUSIONS: Our experience is encouraging if patient selection is accurate and rigid. PMID- 9730397 TI - Gastrointestinal complications after vertical banded gastroplasty. AB - BACKGROUND: Gastric surgical procedures for morbid obesity may have occasional serious complications. The vertical banded gastroplasty according to Mason's technique (VBG) is a common procedure for morbid obesity. The aim of this study is to present the complications in the gastrointestinal (GI) tract after VBG and to discuss their treatment. METHODS: In this study 260 morbidly obese patients (62 males and 198 females) underwent VBG. RESULTS: Complications in the GI tract were encountered as follows: narrowing of the communicating lumen of the two parts of the stomach in four patients, dehiscence of the vertical stomach staple line in three patients, cholelithiasis in 12 patients, gastric perforation in four patients, postoperative fistulas in three patients, serious hepatic failure in one patient, significant gastritis and esophagitis in 32 patients, intestinal obstruction in five patients and frequent prolonged vomiting in 23 patients. The authors attempted to treat all these complications conservatively. In 22 patients, however, a new procedure was necessary. In some cases a partial gastrectomy was necessary. CONCLUSION: VBG is considered to be a satisfactory procedure for weight loss in morbid obesity, but has occasional complications from the GI tract, besides the complications from the other systems. Thus, this procedure should be performed only when absolute indications exist. PMID- 9730398 TI - Converting vertical banded gastroplasty to a lesser curvature gastric bypass: technical considerations. AB - BACKGROUND: Vertical banded gastroplasty (VBG) is occasionally followed by poor weight loss or complications requiring reoperation. Several studies have analyzed the morbidity and mortality associated with conversions of VBG to gastric bypass, but few have described the actual technique. The most frequent complications related to this type of reoperation are gastrointestinal leaks. MATERIALS AND METHODS: The authors analyzed 60 consecutive conversions from VBG to lesser curvature gastric bypass, performed on 60 patients. The cases were analyzed for surgical technique, complications and weight loss. In all the cases the operation was limited to the lesser curvature of the stomach, and certain technical maneuvers were done to facilitate the creation of the pouch and anastomosis. RESULTS: There were three major complications, and two patients required reoperation. There were no gastrointestinal leaks or mortality. Percentage weight loss at 5 years was similar to primary gastric bypasses. CONCLUSION: Converting failed or complicated VBGs to lesser curvature gastric bypasses are safe and effective weight loss operations. By performing several specific technical maneuvers and limiting the operation to the highly vascular lesser curvature, complications can be reduced to a minimum. PMID- 9730399 TI - The left subcostal incision revisited. AB - BACKGROUND: Since 1979, this author has used exclusively a left subcostal incision for gastroplasty (GP) and Roux-en-Y gastric bypass (RYGBP), with complication rates better than published reports in the literature for midline incisions. METHODS AND RESULTS: From July 1979 through March 1997, 1798 primary GP and RYGBP procedures have been done through the left subcostal incision, in addition to 42 conversions of GP to RYGBP, for a total of 1840 new left subcostal incisions. Comparison with an earlier series revealed no significant changes in results: incision hernias three (0.16%), dehiscence four (0.2%), splenectomy three (0.16%). No splenectomies have been necessary since 1983. Various wound healing problems occurred rarely (2.2%). CONCLUSION: The author believes that the left subcostal incision should be the gold-standard of bariatric surgery open procedures. PMID- 9730400 TI - Standards of excellence for bariatric surgery: great concept, but how? PMID- 9730401 TI - VBG: Marlex vs Dacron banding. PMID- 9730402 TI - Vertical pouch ulcers. PMID- 9730403 TI - Combining stories and numbers: a methodologic approach for a critical nursing science. AB - The critical paradigm is increasingly being recognized as an appropriate perspective for the development of nursing knowledge. While different research approaches including feminist, neo-Marxist, and participatory research have been described, all share the goals of empowerment, emancipation, and change. As a relatively new world-view for nursing, the concept of a critical nursing science faces much the same resistance as the interpretive paradigm did a decade ago. This article reviews the aims and assumptions of the critical paradigm; discusses the merits of combining stories and numbers for the agenda of change; and, using examples from our research, describes three strategies for combining stories and numbers in the critical paradigm. PMID- 9730404 TI - On "being inspired" by Husserl's Phenomenology: reflections on Omery's exposition of phenomenology as a method of nursing research. AB - The impact of Omery's article, "Phenomenology: A Method for Nursing Research," on nursing science is appraised. In particular, the influence of her emphasis on "being inspired" was compared with that of her detailed reviews of psychological phenomenologic methods. The author's experience of "being inspired" by Husserl's book, Ideas, is described. The author also discusses the tapping of this resource during three phases of her development as a researcher: (1) appraising methods derived from Husserl's phenomenology; (2) spelling out an approach, with help; and (3) "making clearer while glancing-toward." Omery's proposed linkage between philosophic inspiration and methodologic development is highlighted as a challenge to nurse researchers. PMID- 9730405 TI - Nurses who run with the wolves: the power and caring dialectic revisited. AB - The dialectic of power and caring is illustrated through examples of nurses' experiences reported in a recent study of public health nursing. Examples of caring at each level of the dialectic are provided and implications for nurses and their practice discussed. Nurses whose practice was typically one of empowered caring "ran with the wolves," that is, they demonstrated a connection with their nursing legacy that gave them strength and creativity to imagine transformative possibilities. PMID- 9730406 TI - An emancipatory impulse: a feminist postmodern integrated turning point in nursing research. AB - This article critiques the current debates regarding feminism, postmodernism, and feminist/postmodernism within nursing research. The critique classified the debates into three identifiable constructs: dissatisfaction, fragmentation, and integration. The authors propose a solution from the integration debate as an emancipatory "workable" turning point for nursing research. The solution is situated within feminism and draws on the epistemological constructs of both modernism and postmodernism. Nursing research is framed within the proposed ontological links that characterize the integrated solution. PMID- 9730407 TI - Womanism: a methodologic framework for African American women. AB - Although nurse scholars have become increasingly engaged in feminist research and theory development, only a few have included important feminist thoughts expressed by African American womanist theorists. This article presents an abbreviated review and synthesis of Afrocentric ways of knowing, which includes Black feminist, womanist, and Afrocentric perspectives. A developing methodology for use with African American women is also described. PMID- 9730408 TI - A commentary on Newman's theory of health as expanding consciousness. AB - This is an explication of Newman's theory of health. The authors provide concrete examples of its application and note its points of intersection with certain aspects of Eastern thought and alternative medicine. Ideas from other disciplines are brought in order to illuminate Newman's theory from a variety of perspectives. Certain aspects of the theory the authors find problematic also are noted. PMID- 9730409 TI - Fear of contagion: a response to stress? AB - Most of the research on nurses' responses to people affected by AIDS has comprised atheoretical surveys or cross-sectional correlation studies. A decade ago, Meisenhelder and LaCharite were among the first to propose a theoretical explanation for these responses, including implications for altering them. They posited that "fear of contagion [is] an affective response of the stress-coping process ... [and] highlighted origins of the fear, behavioral manifestations, and avenues for exploration to decrease this perceived threat." This article reexamines the interpretation of the empirical data on which their proposition rested, places that data in the context of other research about the nurses' AIDS care attitudes, including Meisenhelder's own subsequent research, and discusses the model's utility for anticipating and influencing nurses' behavioral response to HIV-affected populations. PMID- 9730410 TI - Foreword: advances in rectal cancer treatment: setting the scene. PMID- 9730411 TI - Embryology and anatomy of the rectum. AB - Rectal cancer surgery is difficult due to the rectum's relatively inaccessible pelvic position and its direct relation to many vital structures. The surgeon is challenged to restore intestinal continuity while working in a confined space. Despite the importance of these issues, the embryology and surgical anatomy of the rectum have been poorly understood. In recent years, cadaver dissections and operative resection under direct vision have provided a clearer picture of the structure of the rectum and mesorectum, their innervation, blood supply, and surrounding structures. New imaging techniques will shed further light on the anatomy of these structures and their anatomic variations. PMID- 9730412 TI - Rectal imaging and cancer. AB - Rectal imaging has evolved substantially during the past 25 years and now offers surgeons exquisite anatomic detail and physiologic information. Dynamic cystoproctography, helical computed tomography, endoscopic ultrasonography, endorectal magnetic resonance imaging, and immunoscintigraphy have become standards for the diagnosis of rectal disease, staging of neoplasia, and survey of therapeutic results. The indications, limitations, and relative costs of current imaging methods are reviewed, and advances in imaging technology that promise future benefits to colorectal surgeons are introduced. PMID- 9730413 TI - Mesorectal excision for rectal cancer: a view from Europe. AB - The local recurrence rate after rectal cancer surgery is discussed as related to conventional and total mesorectal excision (TME) techniques. Studies now show that the wide variation in results between centers and among surgeons depends, at least in part, on differences in surgical technique. We conclude that local tumor recurrence rate is lower after TME than after conventional surgery and emphasize the importance of a standardized macroscopic evaluation of the resected specimen. Population-based registration to evaluate the quality of surgery is recommended. It is also suggested that randomized studies on adjuvant treatment for rectal cancer should include a "surgery only" arm when a local tumor recurrence rate of 10% or less is being studied. Until such investigations are performed, we conclude that the role for adjuvant treatment is questionable and that TME surgery is preferred as the treatment option for Stage T1-T3 rectal cancers. PMID- 9730414 TI - Total mesorectal excision in the treatment of rectal cancer: a view from the USA. AB - The technique of total mesorectal excision (TME) has sparked much controversy in the surgical community with its reported advantages of reduced local recurrence, improved survival, and reduced need for adjuvant therapy. TME is the total excision of the tumor with precise, sharp dissection to include midrectum and integral mesentery of the hindgut which envelopes the midrectum. In a compiled series of over 10,000 patients treated for rectal cancer, the local recurrence rate for surgery alone was 18.5%. Proponents of TME report local recurrence rates from 3.5% to 7.3% and survival rates greater than 80% at 5 years. Histopathological studies suggest that a proportion of patients will be at increased risk of local recurrence if adequate circumferential and distal mesorectal margins are not achieved as proposed in TME. Unlike any other cancers, there appears to be a great deal of surgeon variability in the treatment of rectal cancer, and local recurrence rates range from 5% to 30%. There is an unanswered question about the high rate of recurrence with the abdominoperineal resection where the principals of TME are followed and a wide excision is performed. Further questions concerning TME include clinical function, anastomotic dehiscence, sexual function, and adjuvant therapy. There are also detrimental functional costs of more distal anastomoses as required by TME, and further, more distal anastomoses are associated with increased leak rates and potentially increased morbidity and mortality. In the hands of its proponents, TME has commendable results and achieves outcomes superior to others that use combined surgery and adjuvant therapy. Unfortunately, experienced surgeons using apparently similar dissection techniques have not been able to reproduce such good results. Potential explanations include variations in technical expertise, patient and tumor selection bias, and differences in the extent of follow-up. The functional costs, increased anastomotic leak rates, and increased need for diversion must be weighed against potential reduced local recurrence rates in patients with mid and upper rectal cancers. PMID- 9730416 TI - Transanal treatment of rectal cancer: ablative methods and open resection. AB - Conservative surgical techniques are an alternative to radical surgery for selected patients with rectal carcinoma. The goals of conservative management are to select patients with low risk for nodal metastases and achieve local tumor control while preserving anal sphincter function. Patient selection is critical to achieve this outcome because properly selected patients can obtain results comparable to radical surgery. Selection is based on preoperative histologic characteristics and endorectal ultrasonography. Predictors of pelvic lymph node metastasis risk include tumor grade, depth of penetration, mucinous features, and vascular and lymphatic invasion. Endorectal ultrasound (ERUS) is important in accurately staging the lesion by identifying both depth of invasion and presumptive nodal status. The options for local therapy reviewed include techniques of full-thickness local excision and ablative procedures including endocavitary irradiation, electrocoagulation, and laser therapy. The techniques of full-thickness transanal excision and endocavitary irradiation are described with results from the University of Minnesota experience. PMID- 9730415 TI - Anastomotic integrity and function: role of the colonic J-pouch. AB - Since the colonic J-pouch with a colo-anal anastomosis was first introduced in 1986, many reports have shown the superiority of this design as compared to a "straight" colo-anal anastomosis. These advantages have been demonstrated in retrospective, prospective, and prospectively randomized reports. Furthermore, these attributes are realized for at least 12 and possibly more than 24 months after surgery. PMID- 9730417 TI - Chemoradiation for rectal cancer: current methods. AB - The options available for the surgeon treating patients with rectal cancer have multiplied over the last decade, allowing varied approaches to the disease for individual patients. The development of effective adjuvant therapy in the form of radiotherapy and chemotherapy has led to exciting results-and yet more questions to be answered. The decision to employ adjuvant therapy has led to the development of better staging modalities to improve patient selection for the various treatment protocols. The basic issue of timing of therapy-preoperative vs. postoperative-remains hotly contested, and good, prospective, randomized trials are needed before the questions can be answered. The utility of preoperative multimodality therapy in the downstaging of tumors to make curative resection or sphincter preservation possible must be examined. Advances in surgical therapy have been significant, and groups have reported excellent results with total mesorectal excision (TME) in patients without the addition of adjuvant therapy. Other important surgical issues include ultralow anterior resections with colo-anal or J-pouch anal anastomosis, and the efficacy of sphincter preservation through local excision of invasive rectal cancers with or without adjuvant therapy. Each of these issues needs further study and will have great impact on the treatment of rectal cancer as further experience is gained. PMID- 9730418 TI - Induction therapy for rectal carcinoma. AB - Induction, or preoperative therapy for rectal carcinoma, is a controversial topic which appears to have clinical utility in a number of distinct settings that range from early stage to locally advanced disease. Common to all treatment scenarios is the intent of reducing the likelihood of recurrence in the pelvis following surgery. An additional and evolving role is a reduction in the extent of surgery following a complete or partial clinical response to the induction regimen. While radiation as a single modality can lead to downstaging and, perhaps, a reduction in local recurrence, combined modality with 5-fluorouracil (5-FU) and radiation appears to have a greater therapeutic benefit that may be further enhanced by the administration of leucovorin. PMID- 9730419 TI - Genetics of colorectal cancer. AB - Recent years have brought great advances in the understanding of the pathogenesis of colorectal cancers. The elucidation of the underlying genetic alterations that produce these cancers has made it possible to broadly categorize this disease into two major types, hereditary and sporadic. The hereditary cancers have been divided further into recognized syndromes, mainly familial adenomatous polyposis and hereditary nonpolyposis colorectal cancer. These syndromes have unique clinical pictures and different prognoses in addition to their different genetic bases. Sporadic cancers have been found to have analogous genetic alterations which propel them along the progression from normal tissue to benign adenoma/dysplasia to malignancy. These advances in genetics can potentially lead to clinically useful advances in detection, treatment, and, ultimately, prevention of colorectal cancer. PMID- 9730420 TI - Prognostic markers in rectal carcinoma. AB - Guidelines from two major organizations have recently supported the use of only the serological marker carcinoembryonic antigen (CEA) for the prognostication and monitoring of patients with colorectal carcinoma. However, in view of the exciting advances made recently in elucidating the molecular and cellular biology of adenocarcinoma of the rectum, the molecules that transform the well-ordered normal rectal epithelium into an invasive adenocarcinoma may yield information about the ultimate behavior of that cancer. Consequently, assessing the expression of molecules within a primary cancer may predict the probability of regional and distant metastasis, response to therapy, and outcome. This review analyzes the current state of intratumoral expression of several molecular markers for the management of rectal cancer and evaluates their potential for defining which patients may undergo rectal sphincter preservation and need adjuvant therapy. PMID- 9730421 TI - Pulmonary hypertension--advances in medical and surgical interventions. PMID- 9730422 TI - Heart transplant coronary artery disease detected by coronary angiography: a multiinstitutional study of preoperative donor and recipient risk factors. Cardiac Transplant Research Database. AB - BACKGROUND: Controversy exists regarding donor and recipient factors that promote the development and progression of coronary artery disease after heart transplantation and the likelihood of coronary artery disease causing death or retransplantation. METHODS: To investigate this issue in a large cohort of patients, we analyzed 5963 postoperative angiograms performed in 2609 of the 3837 patients undergoing heart transplantation at 39 institutions between January 1990 and December 1994. Coronary artery disease was classified as mild, moderate, or severe on the basis of left main involvement, primary vessel stenoses, and branch stenoses. Coronary artery disease was considered severe if left main stenosis was > 70% or 2 or more primary vessels stenoses were > 70% or branch stenoses were > 70% in all 3 systems. RESULTS: By the end of 5 years after heart transplantation, coronary artery disease was present in 42% of the patients, mild in 27%, moderate in 8%, and severe in 7%. Coronary artery disease-related events (death or retransplantation) had an actuarial incidence of 7% at 5 years and occurred in 2 of 3 of the patients with development of angiographically severe coronary artery disease. By multivariable logistic analysis, risk factors for donor coronary artery disease included older donor age (P < .0001) and donor hypertension (P=.0002). By multivariable analysis in the hazard function domain, risk factors identified for the earlier onset of allograft coronary artery disease included older donor age (P < .0001 ), donor male sex (P=.0006), donor hypertension (P=.07), recipient male sex (P=.02), and recipient black race (P=.01). The actuarial incidence of severe coronary artery disease was 9% at 5 years. CONCLUSIONS: Angiographic coronary artery disease is very common after heart transplantation, occurring in approximately 42% of the patients by 5 years. Older donor age, donor hypertension, and male donor or recipient predict earlier onset of angiographic allograft coronary artery disease. Although severe angiographic allograft coronary artery disease occurs in only 7% of the patients at 5 years, its presence is highly predictive of subsequent coronary artery disease-related events or retransplantation. PMID- 9730423 TI - The International Society for Heart and Lung Transplantation grading system for heart transplant biopsy specimens: clarification and commentary. PMID- 9730424 TI - Tacrolimus therapy for persistent or recurrent acute rejection after lung transplantation. AB - BACKGROUND: Because the severity, frequency, and duration of acute rejection have been linked to the risk of chronic allograft rejection, controlling persistent or recurrent acute rejection is paramount. Tacrolimus has been effective in the management of recalcitrant rejection in other solid organ transplants, and the initial experience in lung transplantation has been favorable. In this study, the impact of changing from a cyclosporine-based to a tacrolimus-based immunosuppressive regimen in lung transplant recipients with persistent or recurrent acute rejection was analyzed. METHODS: The incidence and severity of acute rejection were retrospectively analyzed in 14 lung transplant recipients who were switched from cyclosporine to tacrolimus maintenance immunosuppression because of persistent or recurrent, biopsy-proven acute rejection. RESULTS: Recipients had been treated for acute rejection an average of 2.6 times before changing from cyclosporine to tacrolimus, and 3 recipients had a course of OKT3 therapy. Tacrolimus therapy was begun 238+/-180 days after transplantation, and the mean follow-up period on tacrolimus treatment was 330+/-201 days. After the changeover from cyclosporine to tacrolimus, the number of episodes of acute rejection per recipient decreased (4.3+/-2.1 to 0.4+/-0.5; p=.0001), the average histologic grade of rejection receded (1.3+/-0.4 to 0.3+/-0.4; p=.0001), and the incidence of acute rejection declined (2.4+/-1.5 to 0.1+/-0.3 episodes per 100 patient-days; p=.0001). CONCLUSIONS: Conversion from a cyclosporine- to a tacrolimus-based maintenance immunosuppressive regimen is an effective approach for managing persistent or recurrent allograft rejection after lung transplantation. PMID- 9730425 TI - Mycophenolate mofetil versus azathioprine immunosuppressive regimens after lung transplantation: preliminary experience. AB - BACKGROUND: Mycophenolate mofetil reduces episodes of biopsy-proven acute cellular rejection or treatment failure in the first year after kidney transplantation; however, limited data exist regarding the efficacy after lung transplantation. METHODS: In a 2-center, nonrandomized concurrent cohort study (level III evidence), we analyzed the incidence of biopsy-proven acute cellular rejection (International Society for Heart and Lung Transplantation grade > or=A2) and decrement in pulmonary function during the first 12 months after successful lung transplantation. All patients received induction immunosuppression with antithymocyte globulin (< or=5 days' duration), cyclosporine and prednisone, in addition to either mycophenolate mofetil (2.0 g/d) [n=11] or azathioprine (1 to 2 mg/kg per day) [n=11]. RESULTS: During the first 12 months after lung transplantation, the mycophenolate mofetil group experienced significantly fewer episodes of acute cellular rejection than the azathioprine group (0.26+/-0.34 vs 0.72+/-0.43 episodes/100 patient-days [mean+/ SD], p < 0.01; 95% CI for the difference=0.126 to 0.813). The change in forced expiratory volume -1 second [deltaFEV1] (liters) between the 3rd and 12th months after lung transplantation was analyzed for the two treatment groups. For this interval, deltaFEV1 for the mycophenolate mofetil group was +0.158+/-0.497 L vs 0.281+/-0.406 L for the azathioprine group (p < 0.05; 95% CI for difference=+0.0356 to 0.843). During the first year, there was 1 death in each group attributed to bronchiolitis obliterans syndrome with concurrent pneumonia. There were no differences in incidence of cytomegalovirus or bacterial infections between the treatment groups; however, a higher prevalence of aspergillus sp airway colonization in bronchoalveolar lavage fluid was observed for the mycophenolate mofetil group (p < .05). The prevalence of bronchiolitis obliterans syndrome at 12 months was 36% for the azathioprine group vs 18% for the mycophenolate mofetil group (p=NS). CONCLUSIONS: Our preliminary experience with mycophenolate mofetil after lung transplantation suggests a decreased incidence of biopsy-proven acute cellular rejection. Furthermore, less decline in FEV1 after 12 months may suggest a reduced incidence or delayed onset for development of bronchiolitis obliterans syndrome. Prospective randomized trials with low beta error (level I evidence) should be performed to assess the efficacy of mycophenolate mofetil vis-a-vis acute allograft rejection and bronchiolitis obliterans syndrome. PMID- 9730426 TI - European Multicenter Tacrolimus (FK506) Heart Pilot Study: one-year results- European Tacrolimus Multicenter Heart Study Group. AB - BACKGROUND: Tacrolimus (FK506) may represent a major advance in the management of allograft rejection after solid organ transplantation. In August 1994 a European heart transplantation pilot study was initiated to assess the efficacy and safety of tacrolimus when administered exclusively through an oral route. METHODS: Eighty-two heart transplant recipients were randomized to treatment (2:1 ratio) with either tacrolimus- (n=54) or cyclosporine-based therapy (n=28). RESULTS: No significant differences were evident between the two treatment groups in either rejection or survival rates at 1 year. Kaplan-Meier estimates of the freedom from rejection were 26.3% and 18.5%, respectively, for the tacrolimus and cyclosporine treatment groups (p=.444). Survival rates were 79.6% and 92.9% (p=.125). At 3 of the 5 centers, patients received antithymocyte globulin during the immediate postoperative period and fared better than those who did not (with acute rejection-free rates of 49.2% and 26.7% for tacrolimus and cyclosporine, respectively [p=.080], as opposed to 7.1% and 8.3% [p=.965]; patient survival rates of 84.6% and 93.3% [p=.382] vs 75.0% and 92.3% [p=.243]). The overall rates of infection, impaired renal function (31.5% vs 21.4%), and glucose intolerance (7.0% vs 4.3%) did not differ significantly between the tacrolimus and cyclosporine treatment groups. Tacrolimus seemed to possess an advantage with regard to a reduced requirement for antihypertensive therapy (59.5% vs 87.5%, p=.025). CONCLUSIONS: Immunosuppression with oral tacrolimus provides a viable alternative to treatment with cyclosporine, particularly when administered in conjunction with antibody therapy. Further studies are warranted to optimize the administration of tacrolimus in this indication. PMID- 9730427 TI - Single-center randomized trial comparing tacrolimus (FK506) and cyclosporine in the prevention of acute myocardial rejection. AB - BACKGROUND: To compare the efficacy and safety of tacrolimus and cyclosporine in heart transplantation, this single-center, prospective, randomized, open-label clinical trial was undertaken. METHODS: Seventy-three adult patients were randomly assigned at the time of transplantation to receive either tacrolimus (n=43) or cyclosporine (n=30) as the primary immunosuppressant. Ten of the 43 patients in the tacrolimus group received the drug intravenously in the perioperative period; all other patients received only oral tacrolimus. RESULTS: With a mean follow-up of 27 months, patient survival rates (tacrolimus 83%, cyclosporine 81%) were similar. Fewer patients experienced acute rejection in the tacrolimus group (79%) than in the cyclosporine group (100%), but the difference was not statistically significant. The number of infections and dialysis and insulin requirements were similar for the 2 treatment groups, but the proportion of patients requiring multidrug antihypertensive regimens was lower in the tacrolimus group (12.5% vs 50.0% at month 6; p=.025). The interpatient variance in pharmacokinetic parameters in a subset of 10 patients was much higher after the first oral dose of tacrolimus than at steady-state (eg, first-dose time at which maximal concentration is reached (t(max)): 3.5+/-2.5h, steady-state t(max): 2.0+/-0.7h), and patients treated with intravenous tacrolimus (n=13) rather than oral tacrolimus (n=30) reached target concentrations faster and with less interpatient variability (eg, at day 0: 9.72+/-10.9 ng/mL intravenously vs 3.31+/ 8.1 orally). CONCLUSIONS: Tacrolimus was associated with similar efficacy and safety profiles compared with cyclosporine. The higher interpatient variance in absorption associated with oral tacrolimus during the first few days after transplantation would suggest that intravenous tacrolimus should be used during the perioperative period. PMID- 9730428 TI - Comparison of PRA-STAT, sHLA-EIA, and anti-human globulin-panel reactive antibody to identify alloreactivity in pretransplantation sera of heart transplant recipients: correlation to rejection and posttransplantation coronary artery disease. AB - BACKGROUND: Screening pretransplantation recipient sera for percent panel reactive antibodies (%PRA) by an anti-human globulin (AHG) assay may identify recipients who are at risk for graft rejection or development of posttransplantation coronary artery disease. However, the pretransplantation AHG %PRA does not always correlate with the occurrence of graft rejection or coronary artery disease. METHODS: We compared the predictive capacity of the AHG-%PRA with that of an enzyme-linked immunoassay (EIA)-based PRA assay that identifies immunoglobulin G bound to soluble human leukocyte antigen (sHLA) class I molecules from pooled platelets of 240 random donors (sHLA-EIA), and that of an EIA-based assay that detects immunoglobulin G anti-HLA class I antibodies bound to sHLA derived from individual HLA-typed cell cultures (PRA-STAT). The pretransplantation sera from 130 cardiac allograft recipients were comparatively tested and results evaluated. RESULTS: Although AHG-%PRA- and sHLA-EIA-determined PRA results were comparable, neither assay discriminated potential recipients at risk for rejection or coronary artery disease. However, cardiac allograft recipients with pretransplantation PRA-STAT sera > 10% were at risk for (1) graft rejection (77% vs 56%, p < .05); (2) more rejections/recipient (1.9 vs 1.0, p < .02); (3) graft rejection within 30 days (92% vs 38%, p < .001); or (4) development of coronary artery disease (48% vs 23%, p < .05) than recipients with pretransplantation PRA-STAT sera < 10%. CONCLUSIONS: PRA-STAT analysis of pretransplantation sera from potential cardiac allograft recipients may be more clinically informative about HLA alloimmunity and a better predictor of adverse clinical events than either AHG-%PRA- or sHLA-EIA-determined PRA. PMID- 9730429 TI - Adult human cardiomyocytes coexpress vimentin and Ki67 in heart transplant rejection and in dilated cardiomyopathy. AB - BACKGROUND: The aim of this study was to investigate whether adult human cardiomyocytes may reexpress vimentin and whether this is linked to cellular activation. METHODS: Myocardial samples of 81 heart transplant recipients (n=183) and patients with dilated cardiomyopathy (n=10) were investigated by immunohistochemistry with the use of the marker molecule vimentin, the muscle specific protein desmin, and Ki67, a marker for cell activation. RESULTS: Vimentin protein expression in cardiomyocytes was found in 28 samples of transplant recipients and 5 myocardial samples of patients with dilated cardiomyopathy. Coexpression of vimentin and Ki67 was found in 52 of 340 vimentin positive cardiomyocytes. CONCLUSIONS: We suggest that the vimentin/Ki67 coexpression indicates cell activation processes as the result of different growth stimuli. PMID- 9730430 TI - Mixed allogeneic chimerism prevents obstructive airway disease in a rat heterotopic tracheal transplant model. AB - BACKGROUND: Mixed bone marrow chimerism reliably produces donor-specific transplantation tolerance for a variety of solid organ and cellular grafts. We used a rat heterotopic tracheal transplant model for chronic rejection to investigate whether mixed chimerism could successfully prevent obstructive airway disease. METHODS: Mixed allogeneic chimeras were prepared by reconstituting lethally irradiated Wistar-Furth (WF) recipients with a mixture of 5 x 10(6) T cell-depleted syngeneic (WF) and 100 x 10(6) T-cell-depleted allogeneic (ACI) bone marrow cells (ACI + WF --> WF). Mixed chimerism was present in all animals 28 days after bone marrow transplantation. Donor-specific, syngeneic, or major histocompatibility complex (MHC)-disparate allogeneic tracheas were implanted in recipient's omentum and removed for histologic analysis 30 to 150 days after transplantation. RESULTS: At 30 days after implantation, median luminal obstruction grades (0=none, 4=complete) of syngeneic and allogeneic tracheas were 0 and 4, respectively. Donor-specific (ACI) tracheas implanted in chimeric (ACI + WF --> WF) recipients were remarkably free of obstruction (median luminal obstruction grade=0 at 150 days) and had excellent preservation of respiratory epithelium. Third-party F344 tracheas implanted in chimeric recipients developed progressive luminal obstruction (grade 2 at 30 days, grade 3 at 90 days). CONCLUSIONS: Mixed allogeneic chimerism induces donor-specific tolerance and prevents development of the characteristic fibroproliferative obstructive lesion of bronchiolitis obliterans in a rat heterotopic tracheal transplant model. Excellent preservation of tracheal structure and morphology was achieved across major and minor histocompatibility barriers. PMID- 9730431 TI - Outpatient inotropic therapy in heart transplant candidates: should its use influence waiting list priority status? AB - BACKGROUND: The use of outpatient intravenous inotropic therapy in heart transplant candidates is contentious. In addition to concerns about morbidity and mortality rates, the current United Network for Organ Sharing (UNOS) heart allocation system presently grants no waiting list priority status benefit to candidates who receive intravenous inotropic therapy in the outpatient setting (UNOS status 2), whereas identical therapy given in an intensive care unit setting does increase priority status (UNOS status 1). The goal of this study was to determine whether an increase in UNOS waiting list priority status is justified in heart transplant candidates receiving outpatient intravenous inotropic therapy by comparing the waiting list mortality of UNOS status 2 candidates on such therapy with that of UNOS status 2 candidates maintained on oral heart failure agents alone. METHODS: This is a retrospective analysis of the pretransplantation outcomes of heart transplant candidates initially listed as UNOS status 2, comparing 29 candidates receiving intravenous outpatient inotropic therapy (group 1) to 109 candidates maintained on oral heart failure agents alone (group 2). RESULTS: The waiting list mortality was not significantly different between the two groups (group 1=7% vs group 2=20%, p=.18); however, group 1 patients had greater morbidity rates while awaiting transplantation than group 2 patients. A greater percentage of group 1 than group 2 patients clinically deteriorated to UNOS status 1 while awaiting transplantation (45% vs 11%), resulting in more group 1 patients undergoing transplantation overall, (59% vs 33%, p=.01) and more group 1 than group 2 patients undergoing transplantation at a higher priority status, UNOS status 1 (76% vs 33%, p=.003). Group 1 patients had more pretransplantation heart failure admissions (1.2 vs 0.6 admissions/total waiting period, p=.02) and longer hospital stays (26+/-39 vs 8.8+/-16 days, p=.03), spent a greater percentage of their total waiting time hospitalized (7% vs 2%, p=.003), and were more likely than group 2 patients to receive intravenous inotropic therapy during hospitalization (70% vs 25%, p=.001). CONCLUSION: This study suggests that heart transplant candidates who require maintenance outpatient intravenous inotropic therapy represent a subgroup of UNOS status 2 candidates with greater waiting list morbidity, but no greater waiting list mortality than candidates who can be maintained on oral heart failure agents alone. However, the current UNOS heart allocation system provides for this increased illness acuity by assigning a higher priority status when necessary. A larger, prospective study is necessary to determine whether a true difference in waiting list mortality rates exists and if an increase in priority status is justified for UNOS status 2 candidates requiring maintenance inotropic therapy. PMID- 9730432 TI - Selection and outcome of ventricular assist device patients: the Muenster experience. AB - BACKGROUND: Because the number of patients on the waiting list for transplantation is increasing and the stagnation in the number of organs donated has led to a more restrictive listing for transplantation, an increased fraction of patients needs to be bridged mechanically. We examined the hypothesis that selection of these patients with regard to urgency status is critical in determining outcome. METHODS: A cohort of 631 patients referred for transplantation to our center between January 1, 1990, and December 31, 1996, was analyzed. Two hundred ninety-seven patients were listed for transplantation and 157 were given transplantation. Forty-one patients had to undergo ventricular assist device implantation (n=34, Novacor; n=6, TCI Heartmate; n=1, Medos), 39 for bridging to transplantation and 2 for permanent support. Initial transplantation evaluation data were analyzed in 3 subgroups (elective bridging, urgent bridging, emergency bridging) and compared with another and with other patients referred for transplantation, specifically those who did not have to be bridged on the waiting list. RESULTS: Patients who underwent elective or urgent assist device bridging were younger and more compromised than the rest of patients accepted on the waiting list (higher functional class, lower mean arterial pressure, lower cardiac index, lower serum sodium, higher pulmonary capillary wedge pressure). In the elective group, overall survival including perioperative mortality rate was better than in the urgent/emergency group and at least as good as in patients who were stable on the waiting list and did not undergo heart transplantation during follow-up. This should prompt cardiologists and cardiac surgeons to consider assist device implantation earlier. PMID- 9730433 TI - Right ventricular function in orthotopic total atrioventricular heart transplantation. AB - BACKGROUND: Total orthotopic heart transplantation was recently introduced into clinical practice as an alternative technique of orthotopic heart transplantation, adding bicaval and left and right pulmonary vein anastomoses to pulmonary artery and ascending aorta connection (total technique). The conventional technique (ventricular transplantation with atrioplasty) is compared with the total technique with particular emphasis on right ventricular performance. METHODS: Forty-eight mongrel dogs (23 to 31 kg) were used for 12 total and 12 standard orthotopic heart transplantations. Right ventricular (RV) function and atrial systole were analyzed with the use of micromanometry, sonomicrometry, and ultrasonic flow probes (preload-independent RV recruitable stroke work, RVPRSW). Fourier analysis was used to calculate RV power and pulmonary vascular impedance. RESULTS: There was no significant difference in cardiac ischemic and bypass times between the two groups. After transplantation, sinus rhythm was preserved after all total transplantations and after only one standard transplantation; no significant hemodynamic differences were observed. RVPRSW in the total group was conserved after transplantation; however, RVPRSW decreased by 39% (+/-8, p < .05) in the standard group. There was also a significant decrease in the rate of RV filling in the standard group after transplantation, suggesting decreased right atrial function. Pulmonary vascular impedance and RV power output were not significantly different after transplantation between the two groups. CONCLUSIONS: Total atrioventricular transplantation is a feasible alternative and conserves normal sinus rhythm. Ischemic and bypass times were not significantly different when the superior vena cava anastomosis is performed last after the release of the aortic cross-clamp. The insignificant decrease in the rate of RV filling with the use of the total technique suggests conserved RV diastolic function after transplantation with less decreased RV function in the total group. PMID- 9730434 TI - Expression of endothelin-1 and effects of an endothelin receptor antagonist, TAK 044, at reperfusion after cold preservation in a canine lung transplantation model. AB - BACKGROUND: Rapid increase of pulmonary vascular resistance (PVR) early after reperfusion remains a major issue in clinical lung transplantation. A potent vasoconstrictor peptide, endothelin- plays an important role in various pulmonary pathophysiologic conditions and might induce increased PVR. We investigated the expression and influence of endothelin-1, and the effects of an ETA and ETB nonselective endothelin receptor antagonist, TAK-044, at reperfusion after cold preservation in a canine lung transplantation model. METHODS: Left single lung allotransplantation procedures were performed in three groups of animals. In group I (n=5) lungs were preserved for 12 hours; in group II (n=5) lungs were preserved for 18 hours; and in group III (n=6) lungs were also preserved for 18 hours, and TAK-044 (5 mg/kg) was administered just before reperfusion. All donor lungs were flushed and preserved with low-potassium dextran glucose solution at 4 degrees C. RESULTS: Six hours after reperfusion, arterial oxygen tension (mm Hg, inspired oxygen fraction=1.0) was 512.9+/-34.7 in group I, 152.4+/-46.7 in group II, and 509.6+/-29.0 in group III; PVR index (dyne x sec x cm(-5) x m2) was 1130+/-142 in group I, 1820+/-142 in group II, and 1287+/-191 in group III. Plasma endothelin-1 level was elevated significantly, and endothelin-1-like immunoreactivity was found in a variety of pulmonary vascular tissue and was seen less with immunohistochemical evaluation in group II in bronchial tissue. CONCLUSIONS: These results suggest that endothelin-1 is expressed as a result of ischemia-reperfusion injury and may worsen early graft function. TAK-044 is beneficial in protecting the graft from high pulmonary vascular resistance and pulmonary edema during the early posttransplantation stage. PMID- 9730435 TI - The mechanism of ejaculation: the glans-vasal and urethromuscular reflexes. AB - To assist in the understanding of the pathogenesis of the various ejaculatory disorders, 9 healthy male volunteers (mean age 30.4 +/- 4.8 years) were studied. The EMG response of the bulbocavernosus (BCM) and ischiocavernosus (ICM) muscles and the external urethral sphincter (EUS) to ejaculation induced by glans penis (GP) vibration was recorded. The test was repeated with individual anesthetization of the GP, BCM, ICM, and EUS. During ejaculation, the BCM, ICM, and EUS showed a significant increase in the motor unit action potentials. The contractions were rhythmic with a mean duration of the contractile episode of 0.8 s and the noncontractile episode of 0.72 s, and with a total muscle activity of 4.2 s. GP vibration after anesthetization of the GP produced no ejaculation or increased EMG activity of the BCM, ICM, and EUS, GP vibration after individual anesthetization of the BCM or the EUS produced semen emission but no ejection, and GP vibration after ICM anesthetization produced ejaculation (emission and ejection). The results suggest that the ejaculatory mechanism consists of two reflexes: the glans-vasal and urethromuscular. The former seems to bring the semen to the posterior urethra (the emission stage of ejaculation) and the urethromuscular reflex ejects it to the exterior (ejection stage of ejaculation). A dysfunction of these two reflexes would seem to induce ejaculatory disorders, a point that needs further study. PMID- 9730436 TI - Testosterone replacement therapy. AB - The benefits conferred by testosterone replacement therapy are substantial, both in the short term for the eradication of symptoms of androgen deficiency, and in the long term for the prevention of osteoporosis. As with any long-term treatment there are risks that must be considered, but overall the benefits achieved far outweigh potential risk. Ideally, androgen replacement therapy should provide physiological serum testosterone levels, as well as DHT and estradiol levels, and correct the clinical symptoms of androgen deficiency in hypogonadal men. This goal is difficult to achieve because the dose dependency of androgen-dependent physiological processes is not known. Androgen preparations that are currently available do not fulfill all criteria for an ideal androgen replacement therapy. Parenteral testosterone esters are effective, safe, practical, and inexpensive. The transdermal testosterone systems provide an alternative to testosterone esters in selected patients but these preparations are expensive. Ongoing studies are showing the benefits of testosterone replacement therapy in aging men, but there is concern about side effects on cardiovascular system and prostate. Thus, clinical decision regarding testosterone therapy in older men should be better defined. PMID- 9730437 TI - Decreased levels of interleukin-12 are not correlated with leukocyte concentration and superoxide dismutase activity in semen of infertile men. AB - The interleukin (IL)-12 levels and the superoxide dismutase (SOD) activity were determined in the seminal plasma of fertile and infertile men with or without leukocytospermia by ELISA. The IL-12 levels differed significantly among the fertile and infertile groups, with the infertile groups showing lower levels that were not modulated by leukocytospermia. The SOD activity did not differ significantly among the fertile and infertile groups and was not correlated with the IL-12 levels. The positive correlation with sperm concentration suggests that IL-12 may have a direct or indirect role in male fertility/infertility and that its levels are not modulated by the presence of leukocytes in the semen. PMID- 9730438 TI - Sialic acid in semen of normozoospermic men. AB - Good sperm motility in normozoospermic men was associated with high bound and total sialic acid (SA) and pyruvate concentrations in the seminal plasma, as well as high endogenous (sperm) pyruvate concentrations. The presence of sialidase in the seminal plasma was also demonstrated for the first time. The results suggest that SA is metabolized to pyruvate by a N-acetylneuraminic acid (NANA)-aldolase in the seminal plasma and that pyruvate subsequently serves as an energy source for sperm. PMID- 9730439 TI - Multicenter study on reproducibility of sperm morphology assessments. AB - Sperm morphology has always been considered an important tool in evaluating a man's fertilizing potential. The objective of this multicentric study was to evaluate intra- and interindividual variability and between-laboratory variation using the same or different criteria of sperm morphology assessment. Semen samples were obtained from 20 males and 32 smears were made of all samples. Eighty coded smears (4 per patient) were sent to 8 laboratories for morphology assessment. The centers applied different classification systems (strict criteria, WHO 1987, Dusseldorf criteria) and participants were asked to analyze the 80 smears twice, with an interval of 1 week between each participant's two analyses. Intraclass correlations between repeats showed that sperm morphology can be assessed with acceptable within observer reproducibility. Expected increases in imprecision were observed up to coefficients of variation of >30% with decreasing morphology scores, regardless of the classification system used. Agreement in correct classification of samples as normal/abnormal was obtained in 80% of cases. Differences in reproducibility between slides may reflect an important source of heterogeneity due to smear preparation. These results emphasize the importance of external quality control systems to improve the value of sperm morphology assessments in the investigation of the male partner in a subfertile couple. PMID- 9730440 TI - Computer-assisted sperm motion analysis of bovine sperm treated with insulin-like growth factor I and II: implications as motility regulators and chemokinetic factors. AB - The effects of insulin-like growth factors (IGFs) I and II on motility of bovine sperm were examined using a computer-assisted sperm motion analyzer (CASA). The following kinematic parameters were examined: percentage of rapidly moving cells, straight-line velocity , curvilinear velocity, average path velocity, amplitude of lateral head displacement, and beat cross frequency. Sperm were treated with IGF-I (100 ng/mL) or IGF-II (250 ng/mL) and compared to sperm in modified Tyrodes' medium only (control) at 90, 180, and 360 min using CASA. Insulin-like growth factor I and II increased the percentage of rapidly moving cells, straight line velocity, curvilinear velocity, average path velocity, amplitude of lateral head displacement, and beat cross frequency compared to the control treatment. These results indicate that IGFs may be involved in initiation and maintenance of bovine sperm motility. PMID- 9730441 TI - New system for long-term monitoring of sperm motility: EDTA effect on semen. AB - Many drugs act as sperm stimulants and are of clinical value for male infertility. Current research deals with the physiological mechanisms of sperm motility/sperm stimulation and how long the effect lasts. For such a study, long term monitoring of sperm motility becomes essential for traditional semen evaluation. A new system was designed to deal with the microscopic images of semen. Its performance was evaluated by studying the effect of EDTA on sperm motility. EDTA increased sperm curvilinear velocity (Vcl) and straight-line velocity (Vsl) by 31 and 20%. EDTA also prolonged the duration of motility by 68 and 61%, respectively. However, EDTA had less effect on the linearity of forward progression (Lin). The proposed system can analyze semen and does well at monitoring sperm motility for short term and long term. It may be valuable to test the possible role of sperm stimulation for male infertility and assisted reproduction. PMID- 9730442 TI - Effect of actinomycin D and cycloheximide on human sperm function. AB - The effects of actinomycin D and cycloheximide on human spermatozoal function were examined to investigate the potential transcriptional and translational activities of human sperm cell during capacitation/acrosome reaction. Treatment with actinomycin D significantly increased and treatment with cycloheximide decreased the acrosome reaction, and the penetration rates in the human sperm penetration of zona-free hamster oocytes assay (SPA). [35(S)]Methionine got incorporated into 3-9 de novo synthesized proteins present in the head and midpiece regions of the swim-up population of human sperm. Treatment with actinomycin D completely blocked and treatment with cycloheximide slightly reduced the synthesis of proteins. There seem to be some transcriptional and translational activities in the head and midpiece regions of human sperm during capacitation/acrosome reaction. PMID- 9730443 TI - Current status of intermittent androgen suppression in the treatment of prostate cancer. AB - Treatment of advanced prostate cancer by continuous androgen suppression results in excellent short-term response but poor long-term survival. Intermittent androgen suppression (IAS) aims to maintain androgen responsiveness of tumor cells by regular cycles of treatment cessations and tumor regrowth to specific prostate-specific antigen limits. First clinical trials demonstrate consistent responses and improved quality of life in most patients on androgen suppression retreatment for up to five cycles, with mean off-treatment periods of approximately 5 to 16 months. Most patients with metastatic disease exhibit early disease progression or androgen independency under IAS, but a subgroup including patients with metastatic disease respond to a single androgen suppression cycle with off-treatment times for up to 48 months. In conclusion, IAS improves the quality of life in patients with primarily hormone-dependent tumors without adverse effects and seems to be most effective in patients with prostate cancer with asymptomatic biochemical progression and low tumor burden. Patients should be treated within the framework of randomized trials and characterized for survival and prognostic factors associated with response to IAS treatment. PMID- 9730444 TI - In prostatism patients the ratio of human glandular kallikrein to free PSA improves the discrimination between prostate cancer and benign hyperplasia within the diagnostic "gray zone" of total PSA 4 to 10 ng/mL. AB - OBJECTIVES: Human glandular kallikrein (hK2) possesses approximately 80% structure identity with prostate-specific antigen (PSA). Moreover, messenger ribonucleic acid for hK2 and for PSA is expressed in both benign and malignant prostatic tissue. We investigated whether the hK2 serum measurement may improve the detection of prostate cancer (PCa) in patients with total PSA of 4 to 10 ng/mL (diagnostic "gray zone"). METHODS: Blood samples were obtained from 90 consecutive male patients with lower urinary tract symptoms and total PSA values of 4 to 10 ng/mL. Eighty-one patients underwent transurethral resection of the prostate and 6 radical prostatectomy. The patients were divided into two groups: I, patients with PCa (n = 20) and II, patients with benign prostatic hyperplasia (BPH) (n = 70). An "in-house" immunofluorometric assay with analytical sensitivity of 0.01 ng/mL and the functional sensitivity of 0.05 ng/mL (at this level the mean coefficient of variation, calculated from the precision profile based on the assays of serum samples, was less than 20%) was used to determine serum hK2 concentrations. Total PSA, free PSA (ProStatus), and PSA complexed to alpha1-antichymotrypsin (PSA-ACT) were also measured. Free/total PSA, hK2/total PSA, and hK2/free PSA ratios were calculated. RESULTS: The serum hK2 could be detected in all samples and in 76 (84.4%) of 90 samples (PCa, n = 18; BPH, n = 58) at given functional sensitivity level. For these cases the median concentration of hK2 was 0.135 ng/mL in PCa and 0.09 ng/mL in BPH (P < 0.1). The median hK2/total PSA ratio was 2% for PCa and 1.6% for BPH (P < 0.2). The median free/total PSA ratio was 0.122 for PCa and 0.215 for BPH (P < 0.0008) and the hK2/free PSA ratio was 0.139 for PCa and 0.075 for BPH (P < 0.000003). At a 7.2% cutoff, the specificity of hK2/free PSA ratio was 48.2% at 100% sensitivity and increased to 60.3% at 94.4% sensitivity level (the area under the receiver operating characteristic curve was 0.86). In comparison, the free/total PSA ratio at a 25.2% cutoff had a sensitivity of 94.4% and a specificity of 27.6% (area under the curve = 0.76). CONCLUSIONS: hK2 was detected in all sera with total PSA values of 4 to 10 ng/mL. Of particular clinical interest is the finding that the hK2/free PSA ratio had a better specificity without loss of sensitivity for PCa than total PSA or the PSA free/total ratio within the range of 4 to 10 ng/mL total PSA. hK2 in combination with free PSA may offer a new diagnostic means for PCa detection. PMID- 9730445 TI - Serum percent free prostate-specific antigen in metastatic prostate cancer. AB - OBJECTIVES: To define the serum prostate-specific antigen (PSA) isoform profile in patients who have prostate cancer but do not have a prostate gland, that is, men who have had a previous radical prostatectomy (RP) and subsequently persistent disease as evidenced by elevated PSA. PSA can be reliably measured in the serum in two major isoforms: PSA complexed to alpha1-antichymotrypsin and uncomplexed free PSA (fPSA). Multiple investigations have illustrated the usefulness of the free/total PSA proportion (percent fPSA) in differentiating prostate cancer from benign prostate disease in patients who still have their prostate gland in situ. METHODS: Sera were evaluated from 52 men who underwent RP and postoperatively had increased PSA. fPSA and total PSA (tPSA) concentrations were determined using the Abbott AxSYM PSA assays. Percent fPSA was calculated for all patients. RESULTS: Median tPSA was 5.45 ng/mL (range 0.93 to 214.99). Median fPSA was 0.69 ng/mL (range 0.11 to 54.93); the median percent fPSA was 13.3% (range 3.9% to 62.9%). There were 27 (52%) patients with percent fPSA less than 15%, 25 (48%) patients with greater than 15%, and 7 (13%) with greater than 30%. No significant relationship was found between percent fPSA and grade, stage, and severity of disease. Percent fPSA was significantly increased in patients who received hormonal, radiation, or combination treatment versus those who received no treatment (P = 0.02 to 0.0007). CONCLUSIONS: Serum percent fPSA in men after RP with persistent prostate cancer encompasses a wide range of values with no clear stratifying factor or factors. These observations and further serial studies in patients with progressive metastatic disease may be important in determining the mechanism(s) for lower percent fPSA in men with newly diagnosed prostate cancer. PMID- 9730446 TI - Measurement of complexed PSA improves specificity for early detection of prostate cancer. AB - OBJECTIVES: Prostate-specific antigen (PSA) is the most useful of all tumor markers. Although the sensitivity is impressive, low specificity results in a lack of cancer detection in a significant proportion of patients undergoing prostate biopsy. Several recent studies have addressed the need for improved specificity. Of all these approaches, the free/total PSA ratio appears to be the most promising. Given that most circulating PSA is complexed to alpha1 antichymotrypsin, and that this moiety represents a greater proportion of the total PSA in those men with carcinoma, we set out to determine whether complexed PSA would improve specificity in the detection of men with prostate cancer. METHODS: Archival sera were obtained from 300 men, 75 of whom had biopsy-proved prostate cancer. All sera had been previously stored at -70 degrees C for variable periods. An investigative assay for complexed PSA (Bayer) was used. The Tandem-R free and total PSA assays (Hybritech) were used according to the manufacturer's recommendations. RESULTS: Among all patients, specificities for the total PSA, free/total PSA, and complexed PSA alone were 21.8%, 15.6%, and 26.7%, respectively, at cutoffs yielding 95% sensitivity. Similar equivalence or superior performance, in terms of specificity relative to the free/total PSA ratio, was seen at other sensitivity thresholds and other total PSA ranges. CONCLUSIONS: Complexed PSA alone performs better than total PSA or the free/total PSA ratio and obviates the need for a second analyte determination. We believe this marker may offer significant enhancement in PSA testing with significant economic advantages. PMID- 9730447 TI - Laparoscopic management of indeterminate renal cysts. AB - OBJECTIVES: We present our follow-up of patients with indeterminate renal cysts who were initially evaluated laparoscopically. We specifically address those patients discovered to have cystic renal cell carcinoma by laparoscopy and the incidence of tract seeding, local recurrence, and distant metastases. METHODS: Between July 1993 and September 1997, 35 patients with indeterminate renal cysts were evaluated laparoscopically. Under laparoscopic visualization, the cyst was located and aspirated, the fluid was sent for cytology, and the floor of the cyst was biopsied. The tissue was then evaluated immediately by one of our genitourinary pathologists, and an intraoperative decision was made. Four patients were found to have cystic renal cell carcinoma and underwent partial or radical nephrectomy in the same setting. An additional patient had a delayed partial nephrectomy 10 days after laparoscopy as a result of change in the final pathology reading. The patients with malignancy were followed with chest x-ray, liver function tests, abdominal computed tomography (CT) scans, and physical examination every 3 months for the first year and then every 6 months thereafter. The average follow-up was 20.2 months (range 8 to 30). RESULTS: Of the 35 patients evaluated in this manner, 5 (14%) were found to have cystic renal cell carcinoma. There has been no evidence of local recurrence or metastatic disease to date. Physical examinations, chest x-rays, liver function tests, and abdominal CT scans all remain negative. CONCLUSIONS: Initial laparoscopic evaluation of complex cysts can save the patient from undergoing needless open surgery. Laparoscopic biopsy of cystic renal cell carcinoma followed by open surgery does not seem to increase the incidence of peritoneal seeding, tract recurrence, or distant metastases. Although the preliminary results are very encouraging, long term follow-up is clearly necessary. PMID- 9730448 TI - Impact of urometabolic evaluation on prevention of urolithiasis: a retrospective study. AB - OBJECTIVES: To obtain information on compliance to therapy and study its effect on recurrences. Over the past 20 years, a selective therapy protocol has been formed for prevention of urolithiasis recurrence. Many studies have been performed on the effectiveness of this therapy, but compliance has never been examined. METHODS: Data were abstracted from 177 medical records of patients who were seen at the outpatient clinic between 1985 and 1994. At that time, they were advised to follow a specific therapy regimen (high fluid intake, medication, and/or specific diet) on the basis of the outcome of a standardized metabolic evaluation. RESULTS: Thirty-six percent of the study population was still compliant to the prescribed therapy after a mean period of 5.3 years of follow up. Therapy-compliant patients were older and had had more treatments, more lithiasis-related complaints, and more frequent follow-up visits. These characteristics suggest that patients' awareness of their disease might improve compliance. Survival analyses showed that patients can be classified into two groups characterized by the frequency of stone formation: a single stone episode versus frequent periods of stone formation. It appeared that the stone recurrence rate was twice as high among patients with a history of frequent stones compared with patients with a single stone episode, even though compliance to therapy seemed lower in the latter group. CONCLUSIONS: The usefulness of urometabolic evaluation and subsequent therapy advice seems questionable. Compliance to a life long therapy is very low after a relatively short follow-up period. This study also suggests a prognostic classification based on stone rate. PMID- 9730449 TI - Holmium:yttrium-aluminum-garnet lithotripsy efficiency varies with stone composition. AB - OBJECTIVES: To test the hypothesis that holmium:yttrium-aluminum-garnet (YAG) lithotripsy efficiency varies with stone composition. METHODS: Single pulses of holmium:YAG energy were delivered using 272-, 365-, 550-, and 940-microm optical fibers to human calculi composed of calcium oxalate monohydrate (COM), calcium hydrogen phosphate dihydrate (CHPD), cystine, magnesium ammonium phosphate hexohydrate (MAPH), and uric acid. Energy/pulse settings were 0.2, 0.5, 1.0, and 1.5 J. Stone crater width and depth were characterized with reflectance light microscopy. RESULTS: For similar energies overall MAPH yielded the deepest and widest craters. CHPD, cystine, and uric acid yielded craters of intermediate width and depth. COM yielded the smallest craters. Within any given composition, increased pulse energy yielded craters of increased width and depth. CONCLUSIONS: Holmium:YAG lithotripsy efficiency varies with stone composition. The rank order of crater size appears to correlate with thermal threshold for each composition. Increased holmium:YAG energy produces larger craters for all compositions. PMID- 9730450 TI - Sensitivity and specificity of NMP-22, telomerase, and BTA in the detection of human bladder cancer. AB - OBJECTIVES: The recent introduction of novel molecular markers into clinical urology has created a need to evaluate the efficacy and utility of these potential markers. The ideal assay for bladder cancer should be noninvasive, sensitive, specific, and cost-effective. We compared the Matritech nuclear maxtrix protein (NMP)-22 assay, telomerase activity, and the Bard bladder tumor antigen (BTA) assay for the detection of human bladder cancer. METHODS: A single voided urine sample was obtained from patients with hematuria without bladder cancer and from patients with known bladder cancer before any treatment. Approximately 50 to 100 mL of voided urine sample was collected and aliquotted for the various assays. The results were compared to single cytologic results and ultimately to pathologic findings. RESULTS: In 47 patients with bladder cancer, the overall sensitivity was 81% for NMP-22, 80% for telomerase, 40% for BTA, and 40% for cytology. For Ta tumors (n = 31), sensitivity was 81% for NMP-22, 70% for telomerase, 32% for BTA, and 26% for cytology. For T1 or higher stage tumors (n = 13), sensitivity was 82% for NMP-22, 91% for telomerase, 64% for BTA, and 64% for cytology. The remaining 3 patients had carcinoma in situ (CIS). When tumors were stratified by tumor grade, grade I tumors (n = 16) were detected at 69% with NMP 22, 65% with telomerase, 13% with BTA, and 6% with cytology. Grade II tumors (n = 14) were detected at 86% with NMP-22, 72% with telomerase, 36% with BTA, and 36% with cytology. Grade III tumors (n = 14) were detected at 93% with NMP-22, 93% with telomerase, 79% with BTA, and 79% with cytology. Patients with CIS (n = 3) were detected at 67% with NMP-22, 100% with telomerase, 33% with BTA, and 67% with cytology. In 30 patients with hematuria but without bladder cancer, the overall specificity of the assays was 77% for NMP-22, 80% for telomerase, 73% for BTA, and 94% for cytology. CONCLUSIONS: In the population tested, NMP-22 and the telomerase assays gave similar sensitivity and specificity for the detection of bladder cancer, and appear to offer a greater sensitivity than the BTA assay and/or conventional cytology. PMID- 9730452 TI - The in vitro bactericidal effect of microwave energy on bacteria that cause prostatitis. AB - OBJECTIVES: We investigated the in vitro nonthermal effects of microwaves delivered from Prostatron 2.0 on Escherichia coli and Enterobacter cloacae. METHODS: The fingers of powder-free, sterile gloves were ligated, and bacterial solutions were transferred into the remaining area of the glove. The gloves were then sealed using silk ligatures. One set of gloves was subjected to the microwave treatment while another set was placed in a temperature-matched waterbath to act as control samples. The gloves containing the treatment group were taped around the probe, at the site where microwave energy exits the probe. During the treatment period, the temperatures from the urethral probe and the rectal probe were carefully monitored. RESULTS: The mean (+/-SD) energy delivered was 46.6 +/- 9.5 kJ (range 30.0 to 59.5) for the 10 trials on E. coli and colony counts in the experimental microwaved gloves decreased significantly compared with control samples (5.26 +/- 4.5 x 10(5) versus 10.16 +/- 9.3 x 10(5) CFU/mL, P = 0.02). For the experiments on E. cloacae the mean (+/-SD) energy applied was 38.5 +/- 12.5 kJ, and a significant decrease in colony counts of microwaved samples was also observed compared with controls (11.04 +/- 4.8 x 10(5) versus 20.08 +/- 10.1 x 10(5) CFU/mL, P = 0.004). CONCLUSIONS: Microwave energy, delivered from Prostatron 2.0, independent of heat production has an in vitro bactericidal effect on laboratory-cultured E. coli and E. cloacae. PMID- 9730451 TI - Long-term follow-up of an EORTC randomized prospective trial comparing intravesical bacille Calmette-Guerin-RIVM and mitomycin C in superficial bladder cancer. EORTC GU Group and the Dutch South East Cooperative Urological Group. European Organisation for Research and Treatment of Cancer Genito-Urinary Tract Cancer Collaborative Group. AB - OBJECTIVES: To determine long-term efficacy of intravesical mitomycin C (MMC) versus bacille Calmette-Guerin (BCG) in patients with superficial bladder cancer with regard to recurrences and progression. METHODS: Patients with superficial bladder cancer (pTa, pT1, pTis) were treated with intravesical MMC (30 mg, weekly for 4 weeks, and thereafter monthly for 5 months) or BCG (weekly for 6 weeks). RESULTS: Three hundred forty-four patients were eligible (171 in the BCG group, 173 in the MMC group). The median follow-up was 7.2 years. Toxicity was not significantly different between the two treatment groups. Efficacy of the two treatment policies was similar with regard to tumor recurrence. With regard to progression to invasive disease, MMC was more effective than BCG in patients without carcinoma in situ (CIS) (P = 0.006). CONCLUSIONS: We can confirm the conclusions of other studies that intravesical treatment with 30 mg of MMC remains an effective treatment option that can also be used in high-risk patients. Like others, we could not confirm that a 6-week course of BCG is more effective in the prevention of tumor progression. Of the 33 patients with tumor progression after intravesical therapy, 20 died of bladder cancer, confirming that tumor progression after intravesical therapy carries a poor prognosis. In this study the difference in toxicity between BCG and MMC was not significant. When comparing studies with MMC and BCG, differences in treatment schedule and/or patient selection should be kept in mind. PMID- 9730453 TI - Few patients with "chronic prostatitis" have significant bladder outlet obstruction. AB - OBJECTIVES: A high prevalence of significant bladder outlet obstruction has been reported among men diagnosed as having "chronic prostatitis." To evaluate the possibility that case selection may determine this high prevalence, we compared findings in patients referred directly to our Urodynamic Unit with that of patients evaluated in our Prostatitis Clinic. METHODS: The videourodynamics records of 201 men aged 18 to 50 years who presented to the Urodynamic Unit with any lower tract symptoms (irritative and/or obstructive with or without pain) were compared with the findings in 123 Prostatitis Clinic patients. The latter were evaluated for obstruction with flow rates and, if abnormal, by retrograde urethrograms and videourodynamics. RESULTS: Only 37 (18%) of 201 patients referred to the Urodynamic Unit had pain as a significant symptom and might have been diagnosed as having chronic prostatitis. Of these 37 patients, 4 (11%) had definite obstruction, 6 (16%) were equivocal, 6 (16%) were hypocontractile, 1 (3%) had pseudodyssynergia, and 7 (19%) had normal findings. The remainder had abnormalities of bladder filling (hypersensitivity in 11 [30%] and detrusor instability in 2 [5%]). Fewer of the 123 patients with prostatitis had obstruction (definite in 2 [1.6%] and equivocal in 1 [0.8%]) (P = 0.03), 2 (1.6%) had hypocontractile detrusors, and 2 had urethral strictures. CONCLUSIONS: Patients referred to the Urodynamic Unit with lower urinary tract symptoms and pain rarely have bladder outlet obstruction. However, they are significantly more likely to have bladder outlet obstruction than patients referred to the Prostatitis Clinic who can be screened for obstruction by history, flow rate, postvoid residual, and retrograde urethrogram. PMID- 9730454 TI - Prevalence of bothersome genitourinary symptoms and diagnoses in younger men on routine primary care visits. AB - OBJECTIVES: To assess the prevalence of bothersome genitourinary (GU) symptoms in younger men on routine primary care physician visits. METHODS: One hundred six men aged 18 to 50 years were approached to complete a brief, self-administered survey that included the American Urological Association Symptom Index, a benign prostatic hyperplasia (BPH) Impact Index, and additional questions about GU pain and sexual dysfunction and about a history of GU diseases. Men with GU symptoms had their outpatient records reviewed. RESULTS: Of the 101 respondents (mean age 36 years), 50% reported GU symptoms. Of these men, 25% were bothered by their symptoms and 17% wanted to talk about them with their physicians; 22% were worried that their GU symptoms might be due to prostate cancer; 27% of all men reported a history of at least one GU disease and 17% had more than one; 16% of all men had been to a urologist. Chart review for the 51 men with symptoms revealed physician documentation of GU symptoms in only 24% of cases and an abnormal GU examination in 8%. One third of reviewed charts documented a GU problem that visit. A broad spectrum of GU diagnoses was documented; no one cause predominated. Ninety percent of all men reported that primary care physicians should routinely ask younger men GU questions as part of their general healthcare. CONCLUSIONS: The high prevalence of bothersome GU symptoms and diagnoses in younger men suggests that information about the clinical, functional, and quality of life implications of these symptoms needs to be collected in this population. PMID- 9730455 TI - Prostate cancer in the post-transplant population. Urologic Society for Transplantation and Vascular Surgery. AB - OBJECTIVES: We conducted a multicenter retrospective study to determine the results of treatment for prostate cancer in solid organ transplant recipients. METHODS: A retrospective analysis of all patients diagnosed with prostate cancer after organ transplantation at five centers was conducted. Data were obtained by chart review and a multipoint data sheet was used to abstract the data from the patient charts. RESULTS: Eighteen cases of prostate cancer were identified from six institutions. Most (84%) of the cancers were clinically localized at the time of diagnosis. Nine (50%) of 18 patients underwent radical prostatectomy, which was the predominant mode of treatment. Overall survival at a mean follow-up of 32 months was 66%, with a cancer specific mortality of 16%. Mortality was 13% for the 15 patients with localized disease and 33% for the 3 patients with metastatic disease at the time of diagnosis. CONCLUSIONS: Most of the patients with prostate cancer being detected after solid organ transplantation were diagnosed with localized disease. Aggressive therapeutic intervention as in the general (nontransplant) population yields good results and should be pursued. Diligent surveillance for prostate cancer in this population using periodic digital rectal examination, serum prostate-specific antigen, and prostate needle biopsy as needed will ensure earlier cancer detection and allow for definitive therapeutic intervention. PMID- 9730456 TI - Improved predictability of extracapsular extension and seminal vesicle involvement based on clinical and biopsy findings in prostate cancer in Japanese men. AB - OBJECTIVES: The accurate preoperative prediction of the extent of cancer by pathologic examination is essential for choosing the optimal treatment for patients with prostate cancer. Currently available clinical staging methods are not adequate and more precise staging is required. METHODS: Using the log likelihood ratio test and receiver operating characteristic (ROC) curve analysis, preoperative variables, including biopsy pathologic findings, were assessed for predicting final pathologic stage in prostate cancer. A multivariate model for predicting disease organ confinement status was established for easy clinical use. RESULTS: The use of the number of cores with cancer and maximum cancer length in conjunction with the three variables (prostate-specific antigen, clinical stage, and biopsy Gleason score) was found to significantly improve predictability of extracapsular extension and seminal vesicle involvement in clinically resectable (n = 96) and localized prostate cancers (n = 81) (P < 0.05). Areas under ROC curves for the above two parameter sets (five- versus three-variable model) were 0.8395 and 0.7109, respectively, for capacity for extracapsular extension prediction in clinically localized cancer. These values for seminal vesicle involvement were 0.7861 and 0.6927, respectively. The logistic model gave positive and negative predictive values of 73% and 78%, and 64% and 83%, respectively, for extracapsular extension and seminal vesicle involvement in clinically localized cancer at a predicted probability of 0.5 or greater. CONCLUSIONS: The present method may be used to predict non-organ confined prostate cancer with greater accuracy than the previously reported model using three variables. PMID- 9730457 TI - Familial study of prostate cancer in Jamaica. AB - OBJECTIVES: Rates of prostate cancer in Kingston, Jamaica are extremely high (occurring in more than 300 men out of 100,000 in 1989 to 1993). This article addresses the familial aggregation of prostate cancer in Jamaica. Early evidence for familial prostate cancer was found in the Utah Mormon population. Increased risk of prostate cancer in men with a family history of prostate cancer has been consistently observed in subsequent studies. There have been few studies, however, involving black men, who are known to have an overall higher risk of developing prostate cancer. METHODS: Two hundred sixty-three patients with prostate cancer documented by histology were studied. Two hundred sixty-three age matched control patients were used for comparison. Extensive pedigrees were obtained for both patients with cancer and controls. Data on other malignancies including lung, breast, colon, stomach, and uterine were also collected. RESULTS: The patients with cancer and the controls were comparable with respect to age and family size. Thirty patients with cancer had a first degree relative (ie, brother, father, or son) with prostate cancer compared to 15 controls. The odds ratio is 2.1 (95% confidence interval 1.1 to 4.4). Nine patients with cancer had a second degree relative (ie, grandfather, grandson, or uncle) affected compared to 3 controls. The odds ratio is 3.1 (95% confidence interval 0.8 to 17.8). There was no statistically significant difference in the rates of any of the other cancers studied. CONCLUSIONS: Familial aggregation of prostate cancer is clearly evident in black Jamaican men. A man with one first degree relative with prostate cancer is twice as likely as the general population to develop prostate cancer. In addition, there may be a statistical difference in the risk of developing prostate cancer if an individual has one second degree relative affected. PMID- 9730458 TI - National trends in the epidemiology of prostate cancer, 1973 to 1994: evidence for the effectiveness of prostate-specific antigen screening. AB - OBJECTIVES: The use of prostate-specific antigen (PSA) to screen for prostate cancer remains controversial. Although it is still too early to measure directly the effects of PSA screening on mortality, we examined changes in the epidemiology of prostate cancer to determine if there is other evidence of the effectiveness of PSA as a screening tool. METHODS: We examined trends in age at diagnosis, and age-adjusted trends in stage and grade at diagnosis, for 140,936 white and 15,662 African American men diagnosed with prostate cancer from 1973 to 1994 in the National Cancer Institute's Surveillance Epidemiology and End Results data base. RESULTS: We found a significant downward trend in age at diagnosis, concomitant with a downward shift in stage of disease at diagnosis, starting with the advent of the PSA era in the late 1980s. We noted most cancers detected since the PSA era to be moderately well differentiated (International Classification of Diseases of the World Health Organization grade 2; Gleason score 5, 6, 7) and organ confined. Although findings were similar for both whites and African Americans, African Americans experienced a greater increase in poorly differentiated disease than did whites. CONCLUSIONS: Changes in the epidemiology of prostate cancer since the advent of the PSA era are consistent with the introduction of an effective screening test. This is evidenced by an increase in detection of significant prostate cancer in individuals who will likely benefit from treatment. PMID- 9730459 TI - The appropriate lower limit for the percent free prostate-specific antigen reflex range. AB - OBJECTIVES: The ability of percent free prostate-specific antigen (PSA) to distinguish benign from malignant prostate disease has been established within the 4.0 to 20.0 ng/mL total PSA range, but its utility within the less than 4.0 ng/mL total PSA range has not been clearly defined. We undertook this study to determine the lower limit for the percent free PSA reflex range. METHODS: Four hundred seventy-nine men (mean age [+/-SD] 63.2 +/- 9.68 years) met the following criteria: (1) a measurable total serum PSA level of 4.0 ng/mL or less (mean 2.64 +/- 0.050); (2) concurrently measured free PSA and percent free PSA calculated (mean 19.3% +/- 0.59%); (3) a sextant biopsy diagnosed benign (B) (n = 376) or malignant (M) (n = 103), at one institution, within 90 days of serum collection; and (4) no prior history of prostate cancer. We defined the lower limit to be the lowest total PSA value at which (1) percent free PSA distributions differed between benign and malignant cases; and (2) percent free PSA could predict malignant disease. We compared age, total PSA, and percent free PSA differences with the Mann-Whitney U test and analysis of variance, and used univariate logistic regression to determine each variable's predictive value. Other statistical analysis was performed with contingency tables, Fisher's exact test, and linear regression. RESULTS: The lowest total PSA value at which percent free PSA both differed between benign and malignant cases and predicted prostate cancer was 4.0 ng/mL. CONCLUSIONS: The lower limit for the percent free PSA reflex range should be 4.0 ng/mL. PMID- 9730461 TI - Correlation of transrectal ultrasound measurements of prostate and transition zone size with symptom score, bother score, urinary flow rate, and post-void residual volume. AB - OBJECTIVES: To assess the correlation of total prostatic size and prostate transition zone dimensions with various measurements of the severity of bladder outlet obstruction secondary to benign prostatic hyperplasia. METHODS: Prostate specific antigen, creatinine, American Urological Association symptom score, bother score, urinary history, uroflowmetry, and post-void residual urine volume determination was followed by measurement of the prostate gland and transition zone on transrectal ultrasound images in 136 men undergoing systematic prostate biopsies. Patients were divided into five groups based on past urinary tract treatment history and the presence of prostate cancer on the biopsies. The total prostate and transition zone dimensions, as well as calculated prostate and transition zone volumes, were compared by Pearson correlation with both the subjective and objective voiding parameters in each patient group. RESULTS: The transition zone dimensions correlated positively with American Urological Association symptom score, bother score, and post-void residual urine volume and correlated negatively with maximum and mean flow rates, particularly in patients with no history of prostate surgery, alpha-blocker administration, urinary infections, irritative voiding symptoms, or prostate cancer. CONCLUSIONS: Transrectal ultrasound measurements of transition zone dimensions correlate better than total prostatic dimensions or calculated prostatic or transition zone volumes with the severity of benign prostatic hyperplasia. Of these, the transverse transition zone dimension demonstrated the best correlation; however, this correlation is probably not adequate for clinical utility. PMID- 9730460 TI - An algorithm combining age, total prostate-specific antigen (PSA), and percent free PSA to predict prostate cancer: results on 4298 cases. AB - OBJECTIVES: To (1) determine if patient age and total prostate-specific antigen (PSA) levels could enhance the ability of percent free PSA to distinguish prostate cancer from benign prostate disease within the 4.0 to 20 ng/mL total PSA range; (2) define the probability of prostate cancer based on patient age, total PSA, and percent free PSA; and (3) define a probability cutoff that distinguishes benign from malignant prostate disease. METHODS: The 3773 urologically referred patients with serum PSA values between 4.0 and 20 ng/mL had a sextant biopsy diagnosed as either prostatic carcinoma (1234) or benign prostatic disease (2539) within 60 days of serum specimen collection. We created a logistic regression model, using patient age, total PSA, and percent free PSA, to assign a probability of prostate cancer, and tested the model on an additional data set (525 patients) to calculate sensitivity and specificity. RESULTS: An 18% probability cutoff detected 95% of malignant biopsies and identified 34% of negative biopsies in the validation set. This approach yielded an 11% percentage point increase in specificity over percent free PSA alone. A 20% probability cutoff detected 90% of malignant cases and identified 42% of negative biopsies. CONCLUSIONS: A prostate cancer probability based on age, total PSA, and percent free PSA is more effective than percent free PSA alone in differentiating benign prostate disease from prostate cancer. This model may assist physicians and patients regarding the need for biopsy. PMID- 9730463 TI - Determination of the site of metabolism of total, free, and complexed prostate specific antigen. AB - OBJECTIVES: To determine the site of metabolism of total prostate-specific antigen (tPSA), free PSA (fPSA), and complexed PSA (cPSA). METHODS: A total of 20 male patients, 50 years old or older, having a clinical indication for left and right heart catheterization were enrolled in this study. Selective blood samples were obtained from the infrarenal, infrahepatic, and suprahepatic inferior vena cava, renal vein, hepatic vein, superior vena cava, pulmonary artery, and femoral artery. cPSA concentration was accepted as the difference between tPSA and fPSA concentrations. RESULTS: We found that tPSA and fPSA concentrations in the infrarenal inferior vena cava were significantly higher than in the systemic artery. There was no significant difference between the systemic artery and the infrarenal inferior vena cava for cPSA concentration. Although fPSA concentration decreased significantly across the renal circulation, the decreases in cPSA and tPSA concentrations were statistically insignificant. In the hepatic circulation, we found that tPSA, fPSA, and cPSA concentrations were significantly decreased. No decrease in tPSA, fPSA, and cPSA concentrations were noted across the pulmonary circulation. CONCLUSIONS: Our results indicate that fPSA and tPSA are released into serum from the prostate but the prostate may not have a significant role in complex formation of PSA. In addition, the liver has a significant role in the elimination of tPSA, fPSA, and cPSA. By contrast, the kidneys have a significant role only in the elimination of fPSA. We also found that the lungs did not have a significant role in the elimination of tPSA, fPSA, or cPSA. PMID- 9730462 TI - Comparability of serum prostate-specific antigen measurement between the hybritech Tandem-R and Abbott AxSYM assays. AB - OBJECTIVES: To assess the comparability of the Hybritech Tandem-R and Abbott AxSYM PSA assays in the setting of a hospital laboratory changing methods of PSA assay. METHODS: A total of 115 serum samples were tested simultaneously with both reagent kits. These include samples from patients evaluated for screening, benign prostatic hyperplasia, and follow-up of prostate cancer. RESULTS: The outcomes of the Hybritech Tandem-R PSA test ranged from 0.0 to 48.3 ng/mL with a median value of 2.4 ng/mL (mean 3.48, SD 5.46). The outcomes of the Abbott AxSYM PSA test ranged from 0.0 to 49.33 ng/mL with a median of 2.22 ng/mL (mean 3.82, SD 5.59). The outcomes of the two assays were found to be highly correlated by the Pearson correlation coefficient (r = 0.9942). When samples were divided according to PSA levels of 0.0 to less than 2.5, 2.5 to less than 4.0, 4.0 to less than 10.0, and 10.0 to less than 25.0 ng/mL, the outcomes were also highly correlated in all PSA level ranges (r = 0.9619, 0.8094, 0.9167, and 0.9081, respectively). CONCLUSIONS: The PSA values of the Tandem-R and Abbott AxSYM assays are highly correlated in the PSA level ranges of 0.0 to less than 2.5, 2.5 to less than 4.0, 4.0 to less than 10.0, and 10.0 to less than 25.0 ng/mL. PMID- 9730464 TI - Percent free prostate-specific antigen values in men with recurrent prostate cancer after radical prostatectomy. AB - OBJECTIVES: Patients with prostate cancer may have more of the complexed form of prostate-specific antigen (PSA) in the serum, whereas patients with benign prostatic hyperplasia have less of this complexed form and thus a higher proportion of the free form. However, the molecular basis for the lower percent of free PSA in patients with prostate cancer remains unknown, and considerable overlap in values exists. We examined this hypothesis in men with recurrent or persistent cancer after radical prostatectomy. These men, who have "pure" cancer in that they have no benign elements to their disease, should have very low percent free PSA values. METHODS: Forty-six men with recurrent (persistent) cancer as manifested by rising PSA values (mean [+/-SD] 2.4 +/- 2.5 ng/mL) after radical prostatectomy were available for analysis. Specimens were analyzed with the use of the Abbott AxSYM free and total PSA assays. The Mann-Whitney U test was used to compare percent free PSA values in this recurrent cancer group with values from a previously defined population of 413 men (225 with benign disease and 188 with prostate cancer before prostatectomy). RESULTS: Median values of percent free PSA in the recurrent cancer group (8.4%) were significantly lower than values in the preoperative cancer (11.7%) or benign (17.4%) groups (P < 0.0001 for both comparisons). Among patients in the "pure" cancer group, 30 (65%) had values less than 10%; however, 4 patients (9%) had values from 1 5% to 1 9%, and another 4 (9%) had values of 20% or greater. Pathologically, patients with higher values (15% or greater) had aggressive disease. All patients with values of 20% or greater had evidence of seminal vesicle involvement or nodal disease. CONCLUSIONS: Although most cancers exhibit low values of percent free PSA, a significant proportion of aggressive tumors will demonstrate high values. Until this latter phenomenon can be explained, the widespread use of percent free PSA to distinguish benign from malignant disease or to stage confirmed malignant disease should be approached with caution. PMID- 9730465 TI - Free/total prostate-specific antigen ratio can prevent unnecessary prostate biopsies. AB - OBJECTIVES: To evaluate the ability of free/total prostate-specific antigen (PSA) ratio to improve specificity of prostate cancer detection, compare Diagnostic Products Corporation (DPC) Immulite and Ciba Corning ACS 180 total (t)PSA assay, and define an assay-specific cutoff point and reflex range for DPC PSA ratio (PSAR). METHODS: In a prospective study, 206 men were enrolled with measurement of both assays. Group 1 consisted of 173 men with a suspicion of prostate cancer (PCA). Thirteen men with known PCA (group 2) and 20 men younger than 32 years (group 3) were used as control groups. RESULTS: Our results in group 1 (115 with benign prostatic hyperplasia [BPH], 58 with PCA) revealed a sensitivity of 82.7%, a specificity of 45.2%, and an accuracy of 57.8% for the DPC tPSA assay (cutoff point more than 4.0 ng/mL) within the entire PSA range. tPSA values of the ACS 180 assay were 1.97-fold higher. Within the tPSA gray zone of 2.5 to 10 ng/mL (66 BPH, 23 PCA), specificity and accuracy of DPC tPSA can be improved by using the DPC PSAR (cutoff point less than 19%) from 33.3% to 71.2% and 42.7% to 70.8%, respectively, maintaining the same sensitivity level of 69.6%. CONCLUSIONS: By combining tPSA testing with PSAR within the gray zone, 39.7% (25 of 63) of unnecessary biopsies can be saved, without missing any additional cancers compared with tPSA testing alone. The optimal reflex range for DPC PSAR is 2.5 to 10 ng/mL and the best PSAR cutoff point for biopsy criterion is less than 19% in our high-risk population, with a cancer yield of 34%. Because we still do not have an international PSA standard, it is important to use assay-specific "normal values" and PSAR cutoff points. PMID- 9730466 TI - Management of primary urethral cancer. AB - OBJECTIVES: To determine the best therapeutic approach for treatment of patients with urethral cancer according to tumor location and clinical-pathologic stage. METHODS: A retrospective review of 21 consecutive patients diagnosed with primary urethral carcinoma was performed. Clinical-pathologic staging, treatment modality, and outcome were analyzed. RESULTS: The overall survival rate was 62%. In patients with clinical Stage Ta-2N0M0 tumors, 8 of 9 patients (89%) are free of disease compared to 5 of 12 patients (42%) with Stage T3-4N0-2M0 tumors (P = 0.03). Best treatment outcome for patients with Stage T3 disease or higher was obtained when multimodality therapy (neoadjuvant chemotherapy and radiation therapy with or without surgery) was administered, with a disease-free survival rate of 60%. CONCLUSIONS: Clinical-pathologic stage was a strong predictor of disease-free survival rate. For patients with Ta-2N0M0 tumors, multimodality therapy may not be required. Conversely, best treatment outcomes in patients with T3-4N0-2M0 tumors are obtained by administering a multimodal therapy combining chemotherapy and radiation therapy with surgical resection. PMID- 9730467 TI - Collagen studies for pediatric ureteropelvic junction obstruction. AB - OBJECTIVES: Ureteropelvic junction (UPJ) obstruction is the most common form of urinary tract obstruction in pediatrics. There is controversy regarding the need for early surgical intervention in many patients with apparent neonatal UPJ obstruction. To demonstrate the differences in type and amount of collagen in adult and pediatric UPJ obstruction, collagen studies were performed. METHODS: The experimental groups are 9 pediatric patients with UPJ obstruction and 13 adult patients with UPJ obstruction. Six patients with normal UPJ were assigned as controls for each experimental group. The collagen content of UPJ was quantitated by hydroxyproline analysis. Immunohistochemical staining and Western blotting for collagen types I and III were performed. RESULTS: The collagen content of pediatric UPJ was significantly lower in value than that of adult UPJ (P < 0.05). Immunohistochemical staining revealed that collagen type I was located in interfascicular space and collagen type III was located in intrafascicular space in both age groups. In Western blotting the relative intensity of collagen type III for pediatric UPJ was weaker than that of adult groups. CONCLUSIONS: These results suggest that a larger series of patients should be examined to determine whether quantitative analysis for collagen types I and III would provide some kind of prognostic test for UPJ outcome in pediatric patients. PMID- 9730468 TI - Glans approximation procedure urethroplasty for the wide, deep meatus. AB - OBJECTIVES: To assess the reliability, cosmesis, and complication rate of the glans approximation procedure (GAP). METHODS: We reviewed 37 consecutive GAP urethroplasties performed at the Children's Mercy Hospital in Kansas City, Missouri over a 5-year period, performed by three different pediatric surgeons. All patients selected had a large, deep ventral groove, typically with a wide open urethral meatus. RESULTS: The mean age was 18 months, with a mean follow-up of 28 months. Of the 37, there was one urethrocutaneous fistula that has since been easily repaired with good results. The parents have been very pleased with the results in all cases, with a straight and strong urinary stream. There was one episode of transient erythema, possibly representing an infection, which resolved after 3 days of oral antibiotics. CONCLUSIONS: The GAP is technically easy, reliable, and offers a relatively low complication rate for the repair of a very select group of patients with hypospadias with a deep ventral glanular groove and wide-mouthed meatus. PMID- 9730469 TI - Minimizing hospital length of stay in children undergoing ureteroneocystostomy. AB - OBJECTIVES: Shortening hospital stay yet not compromising quality of care can result in significant cost savings for children undergoing surgical correction of vesicoureteral reflux. METHODS: We reviewed the medical records of pediatric patients who underwent ureteroneocystostomy between July 1995 and July 1997. A total of 43 patients, aged 0.2 to 18 years (mean 5.2) who all received identical postoperative care, except for their pain management and the time of bladder catheter removal, were included in the study. Twenty-three were treated with intravenous ketorolac tromethamine (Toradol); the remaining 20 received narcotics in the immediate postoperative period. The bladder catheter was removed in less than 24 hours in 22 children, and greater than 24 hours in 21. RESULTS: Patients who received ketorolac tromethamine for postoperative analgesia had on average shorter hospital length of stays than those treated with narcotics (1.4 versus 2.5 days, respectively; P < 0.001). The average stay for children whose bladder catheter was removed within 24 hours postoperatively was significantly shorter than those whose catheter was removed after a 24-hour period (1.4 versus 2.4 days, respectively; P < 0.001). There were no reimplantation failures. One child presented 2 days postoperatively with anemia, which did not require transfusion. CONCLUSIONS: Our review demonstrates that ketorolac tromethamine can be used safely and effectively in children for immediate postoperative analgesia, and that its proper use combined with early catheter removal can reduce the length of hospital stay for pediatric patients undergoing ureteroneocystostomy. PMID- 9730470 TI - Duodenal hematoma after retroperitoneal lymph node dissection. PMID- 9730471 TI - Prevesical space hematoma simulating the normal urinary bladder. PMID- 9730472 TI - Transurethral cystotomy repair. AB - We present a novel transurethral technique for cystotomy repair. This method can be used for immediate bladder repair following limited bladder injury and avoids the morbidity associated with laparotomy. PMID- 9730473 TI - Renal and adrenal surgery during pregnancy. AB - Retroperitoneal surgery during pregnancy is rarely indicated. Major considerations are the optimal timing of surgery and the consequences of nonoperative management on mother and fetus. We report on the first partial nephrectomy performed during pregnancy and one nephroadrenalectomy. When indicated, the second trimester is the ideal time to perform nonobstetric surgery during gestation. PMID- 9730474 TI - Keratinizing desquamative squamous metaplasia of the kidney mimicking transitional cell carcinoma. AB - A 54-year-old patient, who had a remote history of radial nephrotomy for nephrolithiasis, presented with a symptomatic renal mass. The lesion was suspicious for malignancy on excretory urography, retrograde pyelogram, and computed tomography. A nephroureterectomy was performed. The histologic sections showed keratinizing squamous metaplasia. PMID- 9730475 TI - Upper urinary tract inverted papillomas. AB - Inverted papilloma is a rare, benign tumor. Only 33 cases to date have been reported to have occurred in the upper urinary tract. These lesions have a male predominance, are difficult to diagnose, and are associated with urothelial malignancy. Because transitional cell carcinoma can present even 8 years after surgery and in other sites within the urinary system, careful follow-up is essential. This article adds 2 new cases and 1 follow-up of a previously reported case from 1990. PMID- 9730476 TI - Primary adenocarcinoma of the urethra with metastasis to the glans penis: successful treatment with chemotherapy and radiation therapy. AB - Primary carcinoma of the male urethra accounts for less than 1% of malignancies in males. A 54-year-old man with primary adenocarcinoma of the urethra with metastasis to the glans penis and lymphadenopathy in the right groin was treated successfully by combined chemotherapy (5-fluorouracil and cis-platinum) and external beam radiotherapy (total dose of 75 Gy). Short-term remission using multimodal approach with penile preservation was achieved in the present case. PMID- 9730477 TI - Large-cell calcifying Sertoli cell tumor of the testis: case report and review of the literature. AB - Large-cell calcifying Sertoli cell tumor (LCCSCT) is a rare sex cord-stromal tumor found predominantly in the pediatric population. This tumor has distinctive histopathologic features and clinical associations. LCCSCT has also been noted in association with the Carney complex, and in patients with Peutz-Jeghers syndrome. The propensity to metastasize is low, and radical orchiectomy has traditionally been the treatment of choice. PMID- 9730478 TI - An in vitro biocompatibility evaluation of double-J stents. AB - OBJECTIVES: For several years, studies performed to estimate in vitro biocompatibility of urinary catheters have been carried out using permanent cell lines. However, for a rational design of the testing procedure, the cell culture model should depend on the material application. We assess the biocompatibility of 13 double-J stents using an in vitro model of normal human urothelial cells (HUC). This article aims to mimic in vitro, on HUC monolayers, the close contact existing in vivo between the urothelium and double-J stents and to evaluate the subsequent effect on these cells. METHODS: Fragments of each stent were deposited into the wells containing confluent HUC, with close contact maintained between the material and the cells. The same procedure with either no material or fragments of latex catheter was undertaken to provide the negative and positive controls, respectively. The contact was maintained for 1, 3, and 8 days. At the end of the incubation period, fragments of stent were removed and cell activity tests were performed (neutral red assay, MTT assay, and cell proliferation). RESULTS: One of the silicone stents is significantly deleterious on HUC as determined by three tests after 8 days of contact. For two copolymers, a tendency to increase cell proliferation was noted. Concerning polyurethanes, we observed significant decreases in HUC viability and cell metabolic activity for five stents after 8 days of contact. All seven polyurethane stents significantly inhibited cell proliferation. CONCLUSIONS: The HUC culture model may be of relevance for the screening of materials intended for use as double-J stents. PMID- 9730479 TI - PSA of 0.5 ng/mL or less and prostate cancer radiation. PMID- 9730480 TI - ProstAsure Index in the detection of prostate cancer. PMID- 9730481 TI - Maternal plasma hypo-osmolality: effects on spontaneous and stimulated ovine fetal swallowing. AB - Fetal swallowing is a major route of amniotic fluid resorption, and thus swallowing activity may alter amniotic fluid volume. Near-term ovine fetal swallowing increases in response to plasma and/or cerebrospinal fluid hypertonicity. As maternal hydration status alters amniotic fluid volume, we hypothesized that maternal plasma hypotonicity may alter fetal swallowing activity. Pregnant ewes (130 +/- 1 d; n = 6) were chronically prepared with maternal and fetal vascular catheters, a fetal esophageal flow probe, and fetal thyrohyoid and nuchal and thoracic esophagus electromyogram electrodes. Spontaneous fetal swallowing and hypertonic saline thresholds for stimulated swallowing were determined prior to and following maternal hypotonicity induced with water loading and intravenous DDAVP (arginine vasopressin V2 agonist). Fetal swallowing thresholds were determined with intracarotid injections (0.15 ml/kg) of increasing sodium chloride concentrations (0.15-1.2 M) at 2-min intervals. Maternal DDAVP infusion significantly decreased mean (+/-SEM) maternal and fetal plasma osmolalities (298 +/- 2-284 +/- 3; 295 +/- 2-278 +/- 3 mOsm/kg, respectively) and sodium concentrations (147.3 +/- 0.4-137.5 +/- 0.9; 142.7 +/- 0.8-133.5 +/- 1.0 mEq/l, respectively), suppressed spontaneous swallowing activity and volume (1.1 +/- 0.2-0.6 +/- 0.1 swallows/min; 0.7 +/- 0.2-0.5 +/- 0.1 ml/min, respectively) and significantly increased the osmotic threshold for swallowing stimulation (0.77 +/- 0.08-1.03 +/- 0.09 M NaCl). We conclude that: (1) maternal, and thus fetal, plasma hypotonicity results in suppression of spontaneous fetal swallowing activity and a decrease in volume swallowed, suggesting that spontaneous fetal ingestive behavior results, in part, from tonic dipsogenic stimulation, and (2) under hypotonic conditions, the intracarotid NaCl injection concentration for swallowing stimulation increases. These results suggest that the reset (lower) maternal plasma osmolality during human pregnancy may serve to minimize fetal ingestive and perhaps arginine vasopressin-mediated antidiuretic responses to acute maternal hypertonicity. PMID- 9730482 TI - Detection of group B streptococcus: comparison of an optical immunoassay with direct plating and broth-enhanced culture methods. AB - The aim of this study was to compare the diagnostic accuracy of an optical immunoassay (STREP B OIA, Biostar) to direct plating and broth-enhanced culture for the detection of group B streptococcus (GBS) colonization of the lower genital tract in pregnant women. GBS cultures from the lower genital tract were obtained in a prospective fashion using a dual swab transport system from patients with risk factors for perinatal GBS infection. One swab was used to inoculate a trypticase soy agar plate with 5% sheep blood (TSA) and then placed in Lim broth. The other swab was used to perform the Strep B OIA. Growth of GBS by either direct plating or broth-enhanced culture was used as the gold standard for determining GBS colonization. Of the 524 women in the study, 90 women had positive cultures (either TSA or Lim broth). The sensitivity, specificity, positive predictive value, and negative predictive value of the Strep B OIA were 47% (42/90), 96% (416/434), 70% (42/60), 90% (416/464). The sensitivity, specificity, positive predictive value, and negative predictive value of the TSA were 61% (55/90), 100% (434/434), 100% (55/55), 93% (434/469). The sensitivity, specificity, positive predictive value, and negative predictive value of Lim broth were 97% (87/90), 100% (434/434), 100% (87/87), and 97% (434/437). The sensitivity of the Strep B OIA to detect light GBS colonization and heavy GBS colonization, as determined by the TSA, was 53% (19/36) and 90% (17/19), respectively. The Strep B OIA and direct agar plate culture appear to be of limited clinical value due to their poor sensitivities. This study also demonstrates the need to use a selective medium such as Lim broth when assessing for GBS colonization of the lower genital tract. PMID- 9730483 TI - Mechanism of arginine vasopressin suppression of ovine fetal lung fluid secretion: lack of V2-receptor effect. AB - Fetal lung liquid production is essential for in utero pulmonary development, and the resorption of lung liquid at birth facilitates neonatal transition. Lung liquid also contributes importantly to amniotic fluid volume. Factors that influence lung liquid production/resorption may, therefore, impact fetal pulmonary growth and development as well as amniotic fluid homeostasis. Arginine vasopressin (AVP) inhibits fetal lung liquid production and facilitates lung liquid resorption. In view of studies administering fetal AVP or the V2 receptor agonist, 1 desamino 8-D AVP (dDAVP) for the regulation of amniotic fluid volume, we sought to determine the impact of AVP V2 receptor stimulation on fetal lung liquid production. Eight near-term ovine fetuses (130 +/- 2 d; term = 145 d) were prepared with hind limb vascular catheters, a bladder catheter, and a tracheal catheter. Each ewe received vascular and amniotic catheters. After 5 days of recovery, the animals were studied for a 2-hr control period and for 2 hr following intravenous dDAVP injection (1 ng/kg). Lung liquid and urine flow rates and plasma electrolyte and osmolality composition were determined at 30-min intervals. To confirm fetal lung liquid suppression in response to AVP, two animals received an intravenous injection of AVP (2 ng/kg) 24 hr after the first experiment. dDAVP administration had no effect on lung liquid production (3.4 +/- 0.4 ml/kg/h), electrolyte concentrations, or osmolality. Fetal urine electrolyte concentrations and osmolality (154 +/- 24-295 +/- 22 mOsm/kg H2O) increased in response to dDAVP, whereas urine flow decreased (9.4 +/- 2.5-4.2 +/- 1.5 ml/kg/h). In the fetuses exposed to AVP, lung liquid production decreased (3.3 1.6 ml/kg/h). As AVP, although not dDAVP, inhibited fetal lung liquid production, these results indicate that AVP effects on lung liquid are likely mediated via the AVP V1 receptor, not the V2 receptor. The use of fetal administration of dDAVP for the regulation of urine production and amniotic fluid volume will not adversely impact lung liquid production. PMID- 9730484 TI - Accuracy of portable glucose meters for rapid determination of amniotic fluid glucose levels. AB - This study was designed to evaluate the accuracy and feasibility of use of three commercially available portable blood glucose meters to measure amniotic fluid glucose(AFG) levels as compared to an accepted laboratory standard. A prospective study of amniotic fluid from 101 consecutive amniocenteses was performed. Glucose concentration in the amniotic fluid was assessed by hexokinase method in our hospital laboratory (control) and by using three portable meters: Advantage (ADV) (Boehringer Mannheim), Glucometer Elite (ELT) (Bayer), and One Touch II Hospital (T-2) (Lifescan). Twenty consecutive amniotic fluid samples were sent to the laboratory in two vials, the first without additive and the second with potassium oxalate to prevent metabolic activity, to assess the effect of cellular metabolism and time delay on amniotic fluid glucose concentrations. Data are reported as mean +/-SE and were assessed by one-way ANOVA. Of the 101 patients studied, 29 were of gestational age > or = 20 wks. The remaining 72 patients were < 20 wks. All three ambulatory meters demonstrated a linear relationship with control (all P < 0.001). Given a slope of almost 1.0 (m = 0.94) and a y-intercept approaching zero (b = 4.3), the OT2 proved to correlate best with control. ELT: (r2 = 0.55, m = 0.79, b = 22.2) and ADV: (r2 = 0.74, m = 1.45, b = 16.9) both overestimated amniotic fluid glucose. When AFG was < 30 mg/dl via laboratory standard, OT2: (r2 = 0.78, m = 1.05, and b = -2.20, P < 0.001), ADV: (m = 1.02, b = 24.1, r2 = 0.12, P = 0.133). The One Touch II Hospital accurately predicted amniotic fluid glucose at the bedside with excellent correlation including with laboratory standard glucose levels < 30 mg/dl. ADV and ELT proved too inaccurate for clinical use. Control samples were not affected by additives or time delay. These findings confirm that AFG determinations can be obtained rapidly with the OT2 meter at the bedside. PMID- 9730485 TI - Prepregnancy body mass index, weight gain during pregnancy, and perinatal outcome in a rural black population. AB - Relationships between body mass index (BMI) and weight gain with perinatal outcome and birthweight were examined. BMI was calculated on 582 consecutive pregnant women who delivered at or >37 weeks gestational age. Statistical analysis was done using Chi-square tests, analysis of variance, and multiple logistic regression. Of those studied, 13% were underweight, 39% normal, 13% overweight, and 35% obese. Obesity was associated with increasing age (P < .01), multiparity (P < .01), previous cesarean delivery (P < .01), previous macrosomia (P = .01), previous fetal death (P = .03), hypertensive disorders (P < .01), gestational diabetes (P = .02), cesarean delivery (P = .03), and neonatal intensive care unit admission (NICU) (P = .01). The underweight group had the most low birthweight (LBW) infants and the lowest mean birthweight. Ideal weight gain occurred in 31%, inadequate weight gain in 34%, and excessive weight gain in 35%. Inadequate weight gain had increased asthma (P < .05), and hyperemesis (P = .03). Women with ideal weight gain had less smokers (P < .01), fetal distress (P < .05), cesarean delivery (P = .02), and preeclampsia (P < .001). The mean birthweight was highest in the excessive weight gain (P < .01). With multivariate analysis, previous LBW, BMI, and tobacco use were significant predictors of LBW. Normal BMI and ideal weight gain in pregnancy is associated with decreased perinatal complications and an optimum birthweight. PMID- 9730486 TI - Spontaneous delivery through the rectovaginal septum and perineal body: an unusual complication of persistent occiput posterior position. PMID- 9730487 TI - Isolated oligohydramnios in the term pregnancy: is it a clinical entity? AB - The aim of this study was to test the hypothesis that isolated oligohydramnios in the otherwise normal term pregnancy does not indicate fetal compromise. Women undergoing labor induction for isolated oligohydramnios between 37 and 41-6/7 weeks gestation were matched by gestational age and parity to women with normal amniotic fluid index measurements who presented in spontaneous labor. Pregnancies complicated by hypertension, diabetes, fetal anomalies, or suspected fetal growth restriction were excluded. The primary outcome variable was route of delivery. Secondary outcomes examined included presence of meconium, acidosis, low Apgar score, and NICU admission. A total of 183 women underwent induction for isolated oligohydramnios. When compared to the control group, neonatal outcome measures did not differ in the group induced for oligohydramnios. However, the women who were induced had significantly more cesarean deliveries (15.8% vs. 6.6%, P < 0.01, odds ratio 2.7). The increased need for operative delivery was not attributable to more fetal distress in the oligohydramnios group. We conclude that isolated oligohydramnios in the otherwise normal term pregnancy may not be a marker for fetal compromise, and induction of labor may not be warranted in most cases. PMID- 9730488 TI - Understanding and using confidence intervals in clinical research. AB - Confidence intervals contain a wealth of clinically relevant information that is not available in the P value and usual significance testing. Numerous articles discuss the hazards of interpreting study results based solely on the P value, raising both practical and philosophical concerns. The general recommendation is that clinical research should not just test hypotheses, but also describe magnitudes of clinical effect. To this end, the confidence interval is a crucial tool in interpreting clinical studies. In this report, we show how one may use confidence intervals to gain further insight into clinical research. For example, by using confidence intervals, one can identify statistically significant results that are clinically imprecise, or conversely, statistically nonsignificant results that are quite precise. In addition, confidence intervals, like the P value, are influenced by sample size. We show how sample sizes that are sufficiently large to test hypotheses may be too small to generate precise estimates of the magnitude of effect. The application and interpretation of confidence intervals are demonstrated through the use of several examples. PMID- 9730489 TI - Transient synchronous intrapartum late deceleration patterns of concordant twins due to maternal-related etiologies. AB - An unusual case is presented in which intrapartum electronic fetal heart rate (FHR) monitoring of concordant monochorionic twins at 39 weeks gestation revealed two separate occurrences of transient synchronous repetitive late deceleration patterns of both fetal heart rates. Both episodes were considered due to correctable maternal-related etiologies. Each of these occurrences resolved immediately with correction of the respective underlying etiologies, and the subsequent FHR tracings were reactive. PMID- 9730490 TI - Chromosome abnormalities in the placenta and spontaneous abortions. AB - Data are presented confirming that placental chromosome abnormalities are more important than fetal chromosome abnormalities in determining fetal loss. Improved methodologies for studying chromosome abnormalities in spontaneous abortion (SAB) are presented that include results for 141 cases (gestational ages 6-24 weeks) with karyotypic study on placental as well as fetal tissue. Experience in two laboratories gave a success rate of >90% of specimens with identifiable placental tissues, an average turnaround time of 10 days for those that produced chromosome results, male-to-female ratio of >50% (indicating no impact of maternal cell contamination), and 30% of cases had chromosome abnormalities. More significantly, two cases (5% of all abnormal cases) showed an abnormal karyotype limited to the placenta. This illustrates the need to examine the placenta in cases of SAB and the importance of technical issues in laboratory studies. PMID- 9730491 TI - Stringent controls in diabetic nephropathy associated with optimization of pregnancy outcomes. AB - To evaluate maternal-fetal outcomes in pregnancies complicated by diabetic nephropathy were evaluated. Nephropathy was defined as proteinuria of >300 mg/24, or albuminuria >300 mg/24 hr in the absence of infection. Twenty-seven pregnant women with variable degrees of diabetic nephropathy were included in the study. Prenatal care included stringent metabolic control and management of hypertension. Fetal and maternal outcomes were obtained by medical record review. There were no fetal deaths. One neonatal death occurred in a fetus delivered at 29 weeks gestation. IUGR and major congenital malformations were observed in 9% of the neonates; 26% of the infants were delivered preterm. Chronic hypertension (77%) and preeclampsia (53%) were common maternal complications; 63% of women required delivery by cesarean section. Successful pregnancy outcomes were achieved in >95% of the women in our population. Modern management of the pregnancy complicated by diabetes has substantially improved the outcome of class F/FR diabetic mothers and their infants. PMID- 9730492 TI - Human obese gene expression: alternative splicing of mRNA and relation to adipose tissue localization. AB - BACKGROUND: The adipocyte-specific protein leptin signals the size of the adipose tissue mass to hypothalamic regions, thereby influencing food intake and energy metabolism. Human obesity is often associated with high leptin levels implying leptin resistance or defective leptin function. Two leptin mRNA species differing only by the presence or absence of a CAG codon encoding glutamine at position 49 of the mature protein arise from alternative splicing owing to two splice acceptor sites immediately following each other at the intron 2 - exon 3 junction. Since glutamine 49 is part of a highly conserved region, we studied possible functional implications of alternative splicing for human obesity. METHODS: We determined, in lean and obese individuals, the relative abundance of both mRNA species in intra- and extraperitoneal adipose tissue in relation to ob gene transcript abundance and plasma leptin levels. RESULTS: Leptin mRNA levels in adipose tissue and concentrations of leptin in plasma were significantly higher in obese subjects than in controls. In both obese and control subjects, leptin mRNA levels were higher in extraperitoneal than in intraperitoneal adipose tissue. Furthermore, leptin mRNA abundance correlated with average fat cell size. In all tissue samples, the predominant ob gene transcript contained the codon for glutamine 49 and the molar ratio of the two leptin mRNA species was similar in patients and controls. No correlation was observed between splice site usage and leptin mRNA abundance or leptin concentration in plasma in our study group. CONCLUSIONS: Differences in the primary structure of leptin due to the presence or absence of glutamine 49 are unlikely to contribute to the apparent 'leptin resistance' commonly observed in obese individuals. PMID- 9730493 TI - Psychosocial correlates of psychopathology in a national sample of the morbidly obese. AB - BACKGROUND: Although several studies have documented the existence of psychopathology in the morbidly obese, there is disagreement as to its extent and nature. The disagreement has been difficult to resolve because earlier studies have tended to use small, regional samples, and diverse, unstandardized approaches to measuring psychopathology. METHODS: To add clarity, the present study utilized an unusually large, national sample, all subjects of which were administered a standardized, comprehensive test of psychopathology (the MMPI-2), an intensive interview concerning psychosocial history, and a medical examination. Subjects' scores on the MMPI-2 were compared to available norms. The psychosocial interview yielded information about families of both origin and reference. Information about comorbidities, medications, and body mass index (BMI) were available from the medical examinations. Multiple regression analyses were performed to determine the variables that best predicted psychopathology and BMI. RESULTS: The percentage of subjects whose MMPI-2 scores exceeded and approached psychopathological levels was in excess of normative expectations. The 1,3,2 pattern of scale scores on this test expresses depressive disorder, with anxiety and somatization features. Regression analyses showed that abuse of the subject and of substances in the family of origin positively predicted, while education and number of children negatively predicted both psychopathology and BMI. CONCLUSIONS: Dysfunctionality in the family of origin may lead to obesity through such regressive coping styles as stress eating, but this can be offset by personal development and nurturance responsibilities. PMID- 9730494 TI - Eating behavior and the experience of hunger following gastric bypass surgery for morbid obesity. AB - BACKGROUND: Numerous different factors may contribute to the varying degrees of success observed following gastric bypass surgery. It is likely that alterations in the subjective experiences of hunger and satiety, as well as behavioral factors, are important. Our aim was to investigate the association of several factors, including qualitative aspects of hunger and satiety, eating patterns, and the emotional valence of different foods, to the weight loss that occurred following obesity surgery. METHODS: A questionnaire covering aspects of hunger, eating and satiety was administered to three groups: (1) a group of people who had undergone gastric bypass surgery with an acceptable weight loss; (2) a morbidly obese group of patients prior to their surgical intervention; (3) a group of people of normal weight. RESULTS: There were significant differences amongst the three groups in scoring on standardized eating disorder scales, in the amount they could eat, and in the experience of hunger. The presurgery, waiting-list group was more receptive to food-related than interoceptive cues when deciding to stop eating. 'Eating styles' also differed across the groups. CONCLUSIONS: It is concluded that changes in specific food-related behaviors and other psychological variables interact with the physical restriction to eating. The relative weighting of other variables needs further exploration. PMID- 9730495 TI - Outcome following bariatric surgery in super versus morbidly obese patients: does weight matter? AB - BACKGROUND: Numerous investigators have attempted to identify prognostic indicators for successful outcome following bariatric surgery. The purpose of this study was to determine whether degree of obesity affects outcome in super obese [>225% ideal body weight (IBW)] versus morbidly obese patients (160-225% IBW) undergoing gastric restrictive/bypass procedures. METHODS: Since 1984, 157 patients underwent either gastric bypass or vertical banded gastroplasty. Super obese (78) and morbidly obese (79) patients were followed prospectively, documenting outcome and complications. RESULTS: Super obese patients reached maximum weight loss 3 years following bariatric surgery, exhibiting a decrease in body mass index (BMI) from 61 to 39 kg/m2 and an average loss of 42% excess body weight (EBW). Morbidly obese patients had a decrease in BMI from 44 to 31 kg/m2 and carried 39% EBW at 1 year. After their respective nadirs, each group began to regain the lost weight with the super obese exhibiting a current BMI of 45 kg/m2 (61% EBW) versus 34 kg/m2 (52% EBW) in the morbidly obese at 72 months cumulative follow-up. Currently, loss of 50% or more of EBW occurred in 53% of super obese patients versus 72% of morbidly obese (P < 0.01). Twenty-six percent of super obese patients returned to within 50% of ideal body weight (IBW) while 71% of morbidly obese were able to reach this goal (P < 0.01). Co-morbidities and complications related to surgery were similar in each group. CONCLUSIONS: Super obese patients have a greater absolute weight loss after bariatric surgery than do morbidly obese patients. Using commonly utilized measures of success based on weight, morbidly obese patients tend to have better outcomes following bariatric surgery. PMID- 9730496 TI - Is bariatric surgery effective in the treatment of the neurological motor deficit syndromes? AB - BACKGROUND: Persons who suffer from neurological motor deficit syndromes, such as multiple sclerosis, postpolio syndrome, cerebral palsy, myotonia or stroke, are at a particular disadvantage if they are also morbidly obese. There appears to be little in the medical literature describing the role of bariatric surgery and the management of these conditions. METHODS: We offer our experience with six such patients. All had variations of the gastric bypass procedure. RESULTS: Weight loss was quite good, in comparison to other morbidly obese patients. All six noted improved function, usually dramatically improved. CONCLUSIONS: Long-term weight maintenance and functional improvement cannot be assessed in this group of patients. One should expect difficulty in making a long-term assessment in the presence of a progressing debilitating disease. Short-term results were good, and we believe that bariatric surgery should be offered to these patients on an individual basis. PMID- 9730497 TI - Stoma adjustable silicone gastric banding versus vertical banded gastroplasty for the treatment of morbid obesity. AB - BACKGROUND: Among gastric restrictive operations, the procedure of choice is still controversial. The aim of this study is to compare the results of two different gastric restrictive procedures: vertical banded gastroplasty (VBG) and stoma adjustable silicone gastric banding (ASGB). METHODS: Between 1991 and 1996, 51 patients were treated surgically for morbid obesity: 27 underwent VBG and 24 underwent ASGB. Preoperative body weight (BW), body mass index (BMI) and percentage of ideal body weight (% IBW) were (mean+/-SD): 145.7+/-45.3 kg; 53.9+/ 15.9 kg/m2; 249.1+/-73.5% respectively in the VBG group. Corresponding figures for the ASBG group were 132.5+/-22.7 kg; 46.9 7.8 kg/m2 and 207.2+/-35.0%. RESULTS: In the VBG group, the median follow-up period was 26 months (range: 7 47). Eighteen months after the operation BW, BMI, % IBW and percentage of excess weight loss (% EWL) were 85.5+/-26.8 kg, 31.9+/-9.8 kg/m2, 145.4+/-43.9% and 74+/ 1% respectively. Complications included incisional hernia (n=1), and bowel obstruction (n=1). One patient died of acute myocardial infarction on the third postoperative day. In the ASGB group, median follow-up time was 19.7 months (range: 18-26). At 18 months postoperation BW, BMI, % IBW and % EWL values were 86.6+/-20.6 kg 30.6+/-6.6 kg/m2 140.6+/-29.3% and 64+/-1% respectively. Gastric wall erosion occurred in two patients and the bands had to be removed. These patients underwent VBG 6 months later. Complications encountered in this group were incisional hernia (n=1), outlet stenosis and reflux esophagitis (n=1), reservoir leakage (n=1) and gastrointestinal bleeding (n=1). Two patients died of pulmonary embolism and acute gastrointestinal bleeding. CONCLUSIONS: Weight reduction was not statistically significant between the two groups. ASGB was easier to perform and less invasive than VBG. PMID- 9730498 TI - History and technical aspects of bariatric surgery in the Czech Republic. AB - BACKGROUND: Central Europe and the Czech Republic are specific in the prevalence of obesity which has increased by 10-40% during the last 10 years. METHODS: In the Czech republic there is 30 years of experience of a comprehensive approach to obesity treatment which includes: dietary treatment; exercise; behavioral modification; drug treatment; and bariatric surgery. Each of these approaches has its place in complex obesity management. Since 1983 bariatric surgery has been established in the Czech Republic for the treatment of morbid obesity. Vertical banded gastroplasty (VBG), gastric banding, laparoscopic nonadjustable and adjustable gastric bandings have been used over the years. Since 1993 laparoscopic gastric banding has been the only method used in our department. RESULTS: The comprehensive approach for obesity treatment in the Czech Republic has resulted in the development of obesity management and research centers, regional obesity units, obesity out-patients clinics and weight reduction clubs. The surgical treatment is a well-established part of this system and the long term results of surgical treatment are acceptable both in terms of weight loss and complication rate. There has been no statistical difference in weight loss results following VBG and laparoscopic gastric banding, but there is a significant decrease in morbidity, and shorter hospital stay associated with laparoscopic gastric banding. CONCLUSIONS: The surgical approach in obesity treatment has an important place in the comprehensive care of obese patients. Laparoscopic gastric banding in the hands of an experienced surgeon is a method with low morbidity, short hospital stay and long-term weight loss results which are fully comparable with the results of other surgical approaches. PMID- 9730499 TI - The First Annual David K. Miller Distinguished lecture. Bariatric surgery and the law. PMID- 9730500 TI - Intimate saboteurs. AB - BACKGROUND: The bariatric patient exists in dynamic relationship with family members and friends who have considerable influence upon the patient and his or her surgical outcome. When family members and friends behave as intimate saboteurs, they attempt to hamper, hurt, or subvert the bariatric patient's goal of achieving and maintaining a healthy body weight. Successful or not, intimate saboteurs provide significant treatment challenges for the patient and the treatment team. METHODS AND PATIENTS: Patient profiles provide examples of intimate sabotage. The psychological construct of Family Systems Theory is used as a plausible explanation for the sabotage of friends and family. CONCLUSIONS: Multidisciplinary professionals treating the bariatric patient must be aware of the critical influence of intimate saboteurs and the tactics they use to sabotage. Treatment guidelines recommended by Family Systems Theory are presented as strategies to mitigate the influence of intimate saboteurs. PMID- 9730501 TI - Maximizing your chances of getting an insurance approval the first time. AB - BACKGROUND: Support staffs for any bariatric surgeon are confronted with daily requests for information, rejections and non responses. Although we are only in control of one-half of the process (with the insurer holding the other cards), there are specific things which can be done to minimize the amount of time spent on each individual claim and maximize your chances of getting it done the first time. METHODS: This paper explores an attorney/obesity rights advocate's various approaches which successfully lead to the overturning of denials by an insurance company or HMO. Implementing such techniques by the surgeon's office may assist some patients in getting approved without having to hire counsel. RESULTS: By standardizing certain repetitively sought information, utilizing existing technology and creating comprehensive checklists, providers can comprehensively process patient claims with an eye toward providing all necessary information from the start. In addition, 'local knowledge' of the propensities of particular insurers must be documented and kept in mind so that inevitable requests for additional information can be minimized. Lastly, a 'crash course' in insurance law may assist your patients' chances to get approved. CONCLUSIONS: Some denials will not be overturned without the assistance of qualified counsel. However, some potential denials can be defeated before they start by carefully documenting files, using technology to provide ample information for the insurance company decision makers, knowing some basic insurance law and by actively seeking your patient's involvement in their claims. PMID- 9730502 TI - Bleeding gastric pouch ulcer after vertical banded gastroplasty: a rare complication. PMID- 9730503 TI - Outcome of gastric restriction procedures: weight, psychiatric diagnoses, and satisfaction. AB - BACKGROUND: Weight losses following bariatric surgery have varied widely, depending on length of follow-up and various pre-surgical characteristics of patients undergoing surgery. METHODS: One hundred thirty one patients had a detailed presurgical psychiatric evaluation. Patients were assessed clinically for 2 years after surgery and at follow-up a mean of 5.7 years after surgery. RESULTS: Mean presurgical body mass index (BMI) was 52.9 kg/m2; therefore, many patients had 'super obesity'. Two-thirds of the patients were located a mean of 5.7 years after surgery. The mean change in BMI at follow-up was 25% and the mean weight loss was 27%. One-third had excellent or good weight outcomes using the Griffen criteria. Five patients had died by follow-up. There was no relationship between age, gender, or fat content presurgically and weight loss at follow-up, although presurgical weight was associated with greater weight loss at follow-up. Weight regain began 2 years after surgery. There was no relationship between the presence or absence of a presurgical psychiatric diagnosis and weight loss at follow-up. There was also no relationship between the presence of a presurgical psychiatric diagnosis and various mental health parameters at follow-up. Satisfaction with the surgery was marginally associated with weight loss but significantly associated with improved mental and physical health. CONCLUSIONS: Mean weight losses were less than have been previously reported with gastric restriction procedures but the follow-up was longer than usually reported and many patients had 'super obesity' prior to surgery. The implications of 'super obesity' for weight loss are discussed. PMID- 9730504 TI - Symptomatic and clinical improvement in morbidly obese patients with gastroesophageal reflux disease following Roux-en-Y gastric bypass. AB - BACKGROUND: Patients who suffer with gastroesophageal reflux Disease (GERD) endure a worsening of symptoms as their weight increases. When medical treatment of this condition in the morbidly obese patients fails, surgical intervention may be indicated. Choosing a procedure which not only helps achieve weight control but which also relieves symptoms and complications of GERD is the goal. We present a review of patients who have undergone Roux-en-Y Gastric Bypass (RYGBP) and related procedures for this disease. METHODS: One hundred eighty-eight patients undergoing surgery for morbid obesity and for GERD in 1992-1996 were contacted by mail or phone. All of these patients had undergone preoperative esophagogastroduodenoscopy to grade the severity of their disease. Their preoperative symptoms were compared to those experienced postoperatively. RESULTS: One hundred thirty patients underwent a RYGBP with modified Hill fundopexy, 22 patients underwent a distal gastrectomy with modified Hill fundopexy, 8 patients underwent distal gastrectomy alone and 28 patients underwent RYGBP alone. There have been no deaths. There were nine surgical complications, eight early and one at 2.5 years postoperation. Follow-up is 4-48 months. The average BMI dropped from 43 to 30.2 kg/m2. Whereas all patients were on some form of medical therapy before surgery, only 14 reported the need for medication postoperatively. CONCLUSIONS: Surgical intervention for weight control and treatment of GERD has been highly successful in our experience both with respect to weight control and to the reduction of reflux symptoms. Depending upon endoscopic and operative findings a RYGBP with or without an antireflux procedure can provide dramatic improvement. Gastrectomy with antireflux modifications is appropriate in selected cases. PMID- 9730505 TI - The impact of small bowel resection on the incidence of stomal stenosis and marginal ulcer after gastric bypass. AB - BACKGROUND: Stomal stenosis (SS) and marginal ulcer (MU) are reported to occur in 9-20% and 2-13%, respectively, of patients undergoing gastric bypass for morbid obesity. It is hypothesized that decreasing tension on the gastrojejunostomy by performing limited small bowel resection (SBR) would decrease ischemia, thereby decreasing the likelihood of SS and MU. METHODS: A retrospective review of 150 consecutive gastric bypass patients operated by one surgeon from 1993 to 1996 was performed. The incidence of SS and MU was compared in patients with and without SBR. RESULTS: The overall rate of SS was 24.0% and that of MU was 9.3%: the incidence of both was 2.0%. The incidence of SS in patients without SBR was 26.9% and with SBR was 19.6%. The incidence of MU in patients without SBR was 8.9% and with SBR was 9.8%. Neither result was statistically significant by Fisher's exact test. CONCLUSION: There is a trend towards a decrease in the incidence of SS in gastric bypass patients with concomitant SBR although this did not reach clinical significance. PMID- 9730506 TI - Silastic ring gastric bypass: results in 64 patients. AB - BACKGROUND: The silastic ring gastric bypass (SRGB) was introduced by Fobi in 1989, in an effort to combine the advantages of the Roux-en-Y gastric bypass with those of the vertical banded gastroplasty, while avoiding the disadvantages of each. METHODS: The results of our first 64 patients who underwent SRGB with a 5.5 cm ring have been reviewed with particular attention to weight loss, short- and medium-term morbidity and patient satisfaction. Most patients have had regular follow-up, and those not seen during the last 6 months were sent a postal questionnaire. RESULTS: The patients included 52 females and 12 males, ranging in age from 23 to 59 years (median age=39 years) at the time of surgery. Median preoperative weight, body mass index (BMI) and % excess weight were 126 kg (range 89-253 kg), 44 kg/m2 (range 36-78 kg/m2) and 113 (range 76-209) respectively. There were no serious postoperative complications and no deaths. Median hospital stay was 7 days (range 5-14 days). Eight patients (13%) are known to have had a staple-line dihiscence. Eighteen patients (28%) have had major difficulties with eating, and in nine (14%) of these the silastic ring has been removed with resolution of the eating problems, but some gain in weight. In the 54 patients with follow-up data at 2 years, median weight was 78 kg (range 55-137 kg), median BMI was 27 kg/m2 (range 20-43 kg/m2) and mean +/- SD % excess weight loss was 69+/-16. After 2 years of follow-up, eight of 54 patients (15%) were unhappy with the results of the procedure. CONCLUSION: SRGB is an effective, safe and well tolerated procedure for achieving weight loss in the morbidly obese. The principal drawbacks relate to staple-line problems and eating difficulties related to the silastic ring. A 5.5 cm ring is probably too small to be ideal. PMID- 9730507 TI - Silastic ring gastric bypass: a comparison of two ring sizes: a preliminary report. AB - BACKGROUND: The silastic ring (vertical) gastric bypass (SRGB) was introduced by Fobi in 1989, in an effort to combine the advantages of the Roux-en-Y gastric bypass with those of the VBG, while avoiding disadvantages of each. We remain unsure of the ideal ring size. METHODS: Sixty-four patients having SRGB between June 1990 and September 1994 had a 5.5 cm ring placed and 24 patients operated between October 1994 and September 1995 had a 6.0 cm ring placed. Weight loss and quality of eating data is compared 12 months after surgery. RESULTS: Median preoperative per cent excess weight was 113 (range 76-209) in the 5.5 cm group and 106 (range 79-196) in the 6.0 cm group. Weight loss was equivalent at 12 months, with median percent excess weight of 33 (range 8-109) and 27 (range 6-81) in the two groups respectively. Quality of eating data appears better in those with the larger ring size. Nine patients with a 5.5 cm ring have subsequently had their ring removed to improve their quality of eating and a further six may require this in the future. One patient with a 6.0 cm ring has had the ring removed and two others may require this be done. CONCLUSION: An SRGB with a 6.0 cm ring achieves equivalent weight loss to one with a 5.5 cm ring, but with better quality of eating, and less prospect of requiring ring removal. However, there remains a small proportion of patients in whom a 6.0 cm ring is poorly tolerated. For this reason a 6.5 cm ring should be tested. PMID- 9730508 TI - Preliminary results of the duodenal switch. AB - BACKGROUND: The duodenal switch (DS), as a modification of the bilio-pancreatic diversion (BPD), is a 'complex' hybrid operation. METHODS: Sixty patients were operated on during the last 3 years. RESULTS: Two patients died early (3.3%); two late deaths occurred at 4 and 7 months, one due to liver failure and the other due to malnutrition and refeeding syndrome (3.57%); three patients required conversions (5.3%). The two early deaths and all the patients who required conversions had a previous vertical banded gastroplasty. Eleven patients had minor liver abnormalities corrected with medications, and one patient had severe diarrhea for more than a year. Eleven female patients have iron deficiency anemia that requires parenteral supplementation. Mean percent excess weight loss was 86% at 2.5 years. CONCLUSIONS: The DS has been, in our experience, an unsafe operation with unacceptably high operative and postoperative mortality. The conversion rate is acceptable. Weight loss, quality of food intake and life have been excellent. Inadequate follow-up can be dangerous if patients fail to report for regular visits. PMID- 9730509 TI - The influence of a new timing strategy of band adjustment on the vomiting frequency and the food consumption of obese women operated with laparoscopic adjustable silicone gastric banding (LAP-BAND). AB - OBJECTIVE: To evaluate the effects of a new timing strategy of band adjustment on the short-term outcome of obese women operated with adjustable silicone gastric banding. SUBJECTS: The outcome of 30 women without binge-eating disorder operated with laparoscopic adjustable silicone gastric banding with a wider intraoperatory band calibration (LAP-BAND) was compared to that of 30 body mass index-matched women without binge-eating disorder previously operated with adjustable silicone gastric banding (ASGB) applied by laparotomy with the usual intraoperatory band calibration. The patients were evaluated 3, 6 and 12 months after surgery. MEASUREMENTS: (1) weight loss; (2) total daily energy intake; (3) percent as liquid, soft or solid food; (4) vomiting frequency; (5) rate of postoperative percutaneous band adjustments; (6) rate of band-related complications. RESULTS: Both the weight loss and the daily energy intake did not differ between patients with LAP-BAND and patients with ASGB. After surgery, the patients with LAP-BAND ate more solid food and less liquid food than the patients with ASGB. Vomiting frequency was higher in patients with ASGB than in patients with LAP-BAND. The total number of percutaneous band adjustments was higher in women with LAP-BAND than in women with ASGB. Band inflation because of weight stabilization was performed in six (20.0%) women with ASGB and in 19 (63.3%) women with LAP-BAND. Neostoma stenosis occurred in one woman with ASGB, but in none of the women with LAP-BAND. One patient with LAP-BAND presented band slippage. CONCLUSIONS: The wider intraoperatory band calibration performed in patients with LAP-BAND did not reduce the short-term efficacy of adjustable silicone gastric banding. This new timing strategy of band adjustment required more postoperative percutaneous band inflations, but it improved the eating pattern of the patients (low vomiting frequency and high intake of solid food). PMID- 9730510 TI - 42-month preliminary follow-up of the silastic ring vertical banded gastroplasty. AB - BACKGROUND: The authors review preliminary experience with silastic ring vertical gastroplasty (SRVG). METHODS: Of 202 patients who underwent SRVG, 191 are more than 3 months postoperation and of these 165 were accessible for review. RESULTS: Pouch volume could not be readily measured. The TA90BN stapler was occasionally difficult to apply exactly at the angle of His. There was one subphrenic abscess, one gastric bleed, and one dehiscence. Vomiting occurred in eight patients who required reoperation: ring removal, three; cholecystectomy, one; conversion to vertical banded gastroplasty, one; conversion to Roux-en-Y gastric bypass, three. There was no mortality. Weight loss has been satisfactory to 42 months. CONCLUSION: SRVG has been a relatively simple operation, with acceptable morbidity and weight loss thus far. PMID- 9730511 TI - Routine preoperative gastrointestinal series. PMID- 9730512 TI - Gastric bypass: standard surgical technique. PMID- 9730513 TI - Guidelines for reporting results in bariatric surgery. Standards Committee, American Society for Bariatric Surgery. PMID- 9730514 TI - Obesity classification. PMID- 9730515 TI - Medical editors trial amnesty (META). PMID- 9730516 TI - Gastric obesity surgery combined with partial ileal bypass for hypercholesterolemia. AB - BACKGROUND: It is unusual for a patient to manifest both morbid obesity and hyperlipidemia. Certain of these individuals may also have a history of hypertriglyceridemic pancreatitis. We report six morbidly obese hypercholesterolemic patients, two with recurrent hypertriglyceridemic pancreatitis, who were managed by concurrent gastric restrictive surgery and a partial ileal bypass operation. METHODS: The first dual procedure was performed on January 6 1992, and the most recent on February 20 1997. Our series consists of two males and four females, with an average age of 35.5 years at the time of surgery. The mean preoperative weight of these patients was 116.8 kg, and the mean BMI was 39.7 kg/m2. The preoperative mean total plasma cholesterol was 5.5 g/l and the mean plasma triglyceride was 30.61 g/l; the two patients with a history of hypertriglyceridemic pancreatitis had plasma triglyceride levels of 33.6 g/l and 65 g/l. RESULTS: The average weight reduction, using available follow-up intervals was 40.3 kg (34.5%), with a mean postoperative BMI of 20.0 kg/m2 (34.8% reduction). The markedly elevated total plasma cholesterol and plasma triglyceride levels were normalized, with a postoperative mean total plasma cholesterol of 1.47 g/l (73.3% reduction) and a concomitant mean plasma triglyceride of 1.63 g/l (94.7% reduction). The two patients with a history of pancreatitis sustained triglyceride reductions of 96.5% and 94.1%, and neither patient has had an episode of pancreatitis following the dual operative procedures. CONCLUSION: We conclude that the combination of a gastric restrictive operation with a partial ileal bypass procedure represents excellent management for patients with both morbid obesity and hypercholesterolemia, especially if the hypercholesterolemia is accompanied by hypertriglyceridemic pancreatitis. PMID- 9730517 TI - Vertical banded gastroplasty: first experience in Russia. AB - BACKGROUND: The first experience of vertical banded gastroplasty (VBG) in the Russian National Research Center of Surgery is presented. METHODS: From November 1992 to October 1996, 24 morbidly obese patients (mean body weight 147.7 kg, BMI 52.1 kg/m2) underwent VBG according to Mason. RESULTS: The early complication rate was 20.8%. The mean excess weight loss (EWL) after weight stabilization (first 12 patients) was 48.0% in the whole group and 53.9% (range 36.0-73.0%) in 10 patients without staple-line disruptions. Significant positive changes in obesity related diseases were noted. Nine of 23 patients presented with incisional hernias some months after operation. CONCLUSION: The impression of VBG is favorable; however, gaining further experience with the standard techniques and increasing the long-term results are necessary. PMID- 9730518 TI - Vertical banded gastroplasty-gastric bypass in Mexican patients with severe obesity: 1 year experience. AB - BACKGROUND: Different surgical alternatives for the treatment of severe obesity have been described. The two most common surgical procedures are the Vertical Banded Gastroplasty (VBG) and the Roux-en-Y Gastric Bypass (GBP). METHODS: This work describes the results seen during the first 12 months after a surgical technique named Vertical Banded Gastroplasty-Gastric Bypass on 221 Mexican patients with severe obesity operated on between March 1993 and August 1996. RESULTS: 73.3% of the patients were female with an average age of 33.4 years with a standard deviation (SD) of 10 years. The initial mean overweight was 62.2 kg (SD = 26.5 kg). The percentage of ideal weight was 202.3% (SD = 39.4%). The initial body mass index (BMI) was 44.9 kg/m2 (SD = 9.1). The average of excess weight loss in a year was 81.2% (SD = 15.6%) and the BMI was lowered to 26.7 kg/m2 (SD = 5.9). An interesting finding was that the greater the initial overweight, the lesser the resulting weight loss (r = 0.57, P < 0.001). CONCLUSIONS: The procedure was fairly easy to perform. The results were excellent in terms of weight loss and postoperative complications. It is an early experience and the long-term results are still inconclusive; regular check-ups should indicate the procedure's long-term effectiveness. PMID- 9730519 TI - Hemodynamic changes during laparoscopic gastroplasty in morbidly obese patients. AB - BACKGROUND: In nonobese patients, peritoneal insufflation has consistently been shown to influence parameters of preload and afterload as well as cardiac output. Obese patients have an abnormal and particular cardiovascular status. The aim of this study was to investigate the hemodynamic changes induced by an increase of intra-abdominal pressure in morbidly obese patients (MOP). METHODS: Standard general anesthesia was administered to 15 informed MOP (body mass index > 40 kg/m2) scheduled for laparoscopic gastroplasty. Hemodynamic parameters were measured by thermodilution through a pulmonary artery catheter and through invasive blood pressure monitoring. RESULTS: CO2 insufflation with an intra abdominal pressure of 17 mmHg caused a significant increase of mean arterial pressure (MAP) (33%, P = 0.005), mean pulmonary arterial pressure (MPAP) (40%, P = 0.001), pulmonary capillary wedge pressure (PCWP) (41%, P = 0.001), and central venous pressure (CVP) (55%, P = 0.001). The increase in diastolic filling pressures could be due to an increase in the filling volume or to a decrease in diastolic compliance. Ventricular volumes were not measured but we speculate that the rise in CVP, PCWP and MPAP is due to an increase in intrathoracic pressure as judged by the increase of pulmonary airway pressure. Stroke volume fell slightly (11%, P = 0.008), because of a reduction in transmural pressure and a fall in effective preload. Cardiac output rose slightly (16%, P = 0.005) because of an increase in heart rate (15%, P = 0.014) probably induced by sympathetic stimulation, which only became fully operative after 15 minutes. CONCLUSIONS: When compared to nonobese patients our obese patients tolerated the pneumoperitoneum surprisingly well, without experiencing fall in cardiac output. The hemodynamic consequences of peritoneal insufflation seem to be different in obese and nonobese patients. PMID- 9730520 TI - The incidence of clinical postoperative thrombosis after gastric surgery for obesity during 16 years. AB - BACKGROUND: Suggested risk factors for postoperative thrombosis such as high fatty acid levels, hypercholesterolemia and diabetes are common in obese patients. METHODS: In a retrospective study, the case records of 328 patients operated for obesity by gastric procedure from September 1977 until December 1993 were analyzed: 253 women and 75 men with a mean age of 38 years and a mean body mass index (BMI) of 44 kg/m2. The operation time, use of epidural anesthesia, and the occurrence of risk factors; fatty acid levels, hypercholesterolemia and diabetes were recorded. Symptomatic thromboses were verified by phlebography or phlethysmography and pulmonary embolism with ventilation/perfusion scintigraphy or autopsy. RESULTS: The mean operating time was 128 minutes, 77% had epidural anesthesia and the mean hospital stay was 12.3 days. The long hospital stay was due to the fact that most patients took part in different scientific studies perioperatively. The incidence of thromboembolism was 2.4%. Four patients had pulmonary embolism, in one of them this was fatal. Three patients had deep leg vein thrombosis and one patient had arm thrombosis secondary to a central venous catheter. None of these patients had high fatty acids, diabetes or high cholesterol. Of the patients, 298 were given dextran-70 (Macrodex, Pharmacia) as prophylaxis, seven were given heparin and 23 were given no prophylaxis. In the patient group without diagnosed thrombosis, 31% had high fatty acid levels, 2% had high cholesterol levels and 9% had diabetes. CONCLUSIONS: Obese patients seem to have a moderate risk of developing postoperative thrombosis when an effective prophylaxis is used. High free fatty acids, hypercholesterolemia and diabetes are not obvious extra risk factors in obese patients. Thromboprophylaxis should be given to all operated obesity patients regardless of age. The surgeons must be aware and investigate promptly any symptoms suggestive of thromboembolism. PMID- 9730521 TI - Weight loss after extended gastric bypass. AB - BACKGROUND: Gastric bypass (GBP) is the most effective method for controlling morbid obesity. Previously we showed that extending the length of the Roux limb increased weight loss. We have done over 400 obesity operations during the past 20 years. The current study consists of patients from the last 10 years of our experience and compares short to extended Roux-en-Y GBP. METHODS: Data from all patients operated at the Ottawa General Obesity Clinic were entered into a database on an ongoing basis, and those from the past 10 years were analyzed. All patients had standardized preoperative investigations and postoperative follow up. Details of these and of the operative technique are provided in the manuscript. RESULTS: The preoperative weight and BMI were 129 +/- 2 kg, and 46 +/ 2 kg/m2, respectively. The mean weight loss prior to surgery was -2 +/- 21 kg. The results were classified, by percentage weight loss as: 'excellent' = > 35%; 'good' = 25-34%; 'poor' = 15-24%; and 'failure' = < 15%. Sixty-five patients (69%) were available for 2-year follow-up. At this time, mean percentage weight loss was 34 +/- 2 versus 40 +/- 1 for the short Roux (45-135 cm) and long Roux (180-225 cm) groups, respectively (P < 0.01). There were no deaths, leaks, splenectomies or intra-abdominal infections. The incidence of hernia and/or reoperation for bowel obstruction was 35/121 or 29%. The overall incidence of diarrhea was 16/121 (13%) and 6/121 (5%) at 12 and 24 months. Quality of life is significantly impaired in at least three of these patients, all with extended limbs. Major vitamin deficiencies, alterations in liver functions, or other metabolic complications did not occur. CONCLUSIONS: Gastric bypass is the procedure of choice for morbid obesity. Weight loss is marginally improved in proportion to the length of the Roux limb but at a risk of diarrhea, which occasionally may not manifest itself for 8 to 12 months. It is important that methods be devised to correct follow-up, incisional hernias and diarrhea. PMID- 9730522 TI - The Swedish Adjustable Gastric Banding (SAGB) for morbid obesity: 9 year experience and a 4-year follow-up of patients operated with a new adjustable band. AB - BACKGROUND: We have developed an adjustable gastric band in which the stoma diameter can be adjusted from the outside. A standardized technique was employed and the application of our band in terms of weight loss and complication rate was evaluated METHODS: Between August 1990 and November 1991, 50 patients (15 men and 35 women) were operated on by laparotomy. Their mean age at surgery was 41 (19 60) years. Mean preoperative weight was 134 (106-181) kg and the mean BMI was 46 kg/m2 (range 33-59 kg/m2). RESULTS: No patient was lost to follow-up. Four were excluded from the study (brain tumor, pregnancy and two reoperations). The remaining 46 were followed for at least 4 years. At follow-up, mean weight was 80 kg and mean BMI was 27.5 kg/m2. The patients had lost a mean of 54 kg. Two patients (4%) had abdominal reoperation because of technical problems. There was one incisional hernia and one minor wound infection, but no other significant complications. CONCLUSION: This relatively simple method appears to be at least as good as the other operations, and weight loss can be adjusted to patient comfort. Currently, the procedure is being performed laparoscopically. PMID- 9730523 TI - Laparoscopic adjustable silicone gastric banding (Lap-Band): how to avoid complications. AB - BACKGROUND: The laparoscopic application of LAP-BAND is gaining widespread acceptance as a gastric restrictive procedure. At the same time the reported morbidities (i.e., gastric perforation, stomach and/or band slippage) are cause for some concern. METHODS: From September 1993 until May 1997, 260 patients underwent LAP-BAND at the Department of Surgery at the University of Padova, Italy. RESULTS: The mortality rate was zero and the morbidity rate requiring reoperation was 3.4% (stomach slippage, gastric perforation, erosion). In order to avoid complications the key points of the technique are reviewed: (1) reference points for dissection (equator of the balloon, left crus); (2) retrogastric tunnel within the layers of the phrenogastric ligament; (3) embedment of the band; (4) proper outlet calibration; and (5) retention sutures. CONCLUSIONS: Attention to technical details is of paramount importance for a safe, standardized and effective operation. PMID- 9730524 TI - Propantheline as an adjuvant to weight loss after vertical gastroplasty. AB - BACKGROUND: Weight loss after gastroplasty surgery is sometimes unsatisfactory despite normal surgical anatomy and no evidence of deliberate dietary indiscretion. METHODS: Two morbidly obese female patients were treated with a modified Long vertical gastroplasty following failed attempts at weight loss using nonsurgical means. One of these (109 kg) complained of a lack of satiety soon after the surgery and her weight loss was unsatisfactory at 11 kg after 90 days. The other patient (128 kg) lost weight as expected but at 3 months leveled off her weight at 93 kg also with an associated loss of satiety. Both patients noted an increased capacity for food and ease of eating solids including red meat. Gastroscopy revealed normal gastric anatomy in both and the patients were prescribed oral propantheline. RESULTS: There was an immediate subjective improvement in satiety, a reduction in capacity for food and an increase in difficulty with eating some solids. There was a resumption of weight loss that has been sustained down to healthy weight range. CONCLUSION: In two patients with unsatisfactory weight loss after gastroplasty but no demonstrable surgical defect, adjuvant propantheline appeared to induce an excellent further weight loss. PMID- 9730525 TI - Bariatric surgery and multiple personality disorder: complexities and nuances of care. AB - BACKGROUND: Multiple personality disorder (MPD) can occur in patients with morbid obesity in need of bariatric surgery, though few reports noting this association exist in the literature. Herein we address MPD in morbid obesity, in the context of a patient presenting to us seeking surgical treatment of her morbid obesity. METHODS: A 31-year-old morbidly obese (BMI 49 kg/m2) Hispanic female presented in early 1994 requesting bariatric surgery. She had been a victim of violent sexual abuse as a young girl. Subsequently, she developed at least three personalities, including one male personality. RESULTS: Although she has lost nearly 45 kg after gastroplasty, her care has been complicated by her named multiple personalities. While MPD are infrequent and unfamiliar to most care providers, successful outcomes can be promoted with a proper approach. CONCLUSIONS: This patient's care illustrates that: (1) all personalities must agree to proposed operative intervention; (2) consent must be obtained from the 'true' patient; and (3) postoperative care and follow-up must address all personalities for an optimal outcome. PMID- 9730526 TI - VBG: Marlex vs Dacron banding. PMID- 9730527 TI - Rediscovering the wheel in obesity surgery. PMID- 9730528 TI - Waking up the gastric bypass patient. AB - Waking up the gastric bypass patient in the post-anesthesia care unit (PACU) is a continual challenge. From January 1992 to November 1996, 961 gastric bypasses (GBP) have been performed at Columbia St Mark's Hospital. Of the 961 patients, 957 came to the PACU. Four patients went directly to ICU because of respiratory status requiring mechanical ventilation. There have been no deaths and no respiratory arrests in PACU. Continuous bedside monitoring of the patient's respiratory status coupled with pain management contributed to positive care of the GBP patient. Methods of care for the GBP patient include the use of O2 masks and cannulas, coughing and deep breathing, administering i.v. narcotics until patient controlled analgesia pumps are initiated, encouragement and emotional support, ongoing assessment of patients' status, and treating problems/needs appropriately. PMID- 9730529 TI - Appetite-suppressant drugs and the risk of primary pulmonary hypertension? PMID- 9730530 TI - A comparison of fat intake of normal weight, moderately obese and severely obese subjects. AB - BACKGROUND: Excess dietary fat has been implicated in the etiology of obesity. METHODS: This study examined the fat intake of three weight groups, normal (20.0 < or = BMI < or = 27.0), moderately obese (27.1 < or = BMI < or = 39.9) and severely obese (BMI > or = 40.0). Each group contained 50 subjects. Detailed 3 day food records were used to gather the nutritional data. Anthropometric and sociodemographic information was also collected. RESULTS: Overall fat intake was 89 +/- 42 g/day or 37 +/- 10% of total energy. Total fat (g/1000 kcalories) intake was found to be significantly higher in the obese groups (p < 0.05). Subjects in the moderately and severely obese groups consumed significantly more fat and cholesterol and less carbohydrate than did normal weight subjects. Compared to the normal weight subjects, obese subjects also had higher intakes of saturated, monounsaturated and polyunsaturated fat (as a percentage of dietary energy). There was no difference in energy or protein intake, and P/S ratio among the three groups. BMI was strongly positively correlated with total fat, saturated, monounsaturated, polyunsaturated fat, cholesterol, and protein intake (as g/day only), and negatively correlated with carbohydrate intake and the CHO/FAT ratio. Energy intake was not significantly associated with BMI. CONCLUSION: A high fat diet may promote obesity, independently of its calorie contribution. PMID- 9730531 TI - The workup for bariatric surgery does not require a routine upper gastrointestinal series. AB - BACKGROUND: Morbid obesity is a serious disease that afflicts over five million Americans, threatening their health with such co-morbidities as diabetes, arthritis, pulmonary failure and stroke. Surgery is the only effective therapy, providing long-term control of weight, diabetes, pulmonary failure, and hypertension for as long as 14 years. Because the operation presents a major expense, this study examined whether X-ray examination of the gut could be omitted safely as a cost-saving measure. METHODS: The records of 814 consecutive morbidly obese patients who underwent gastric bypass were reviewed to determine: (1) whether these individuals had undergone an upper gastro-intestinal (GI) series, and (2) if these studies influenced therapy or caused cancellation or postponement of surgery. RESULTS: Of the 814 patients, 657 (80.7%) underwent a preoperative GI radiography. Of these examinations, 393 (59.8%) were normal, with the following abnormalities in the remaining 264: hiatal hernia, 164; esophageal reflux, 39; Schatzki's ring, 18; small bowel diverticula, four; renal stones, four; malrotation, three; gall stones, two; pyloric ulcer, one; possible pelvic mass, one; calcified leiomyoma, one; and dysphagial lusoria, one. None of these findings resulted in cancellation or a delay in surgery. CONCLUSIONS: The upper GI series can be safely omitted from the routine preoperative evaluation of patients undergoing gastric bypass. At a cost of $741.00 per examination, this change represents significant potential savings. Similar evaluations of other routine preoperative tests may well provide a better basis for the evaluation of these complex patients. PMID- 9730532 TI - Laparoscopic adjustable silicone gastric banding: preliminary results of the University of Naples experience. AB - BACKGROUND: Laparoscopic adjustable silicone gastric banding (LASGB) is a minimally invasive surgical procedure indicated for the treatment of patients with morbid obesity. METHODS: From January 1996, eight patients successfully underwent the video-laparoscopic procedure. RESULTS: Preoperative body mass index was 44.4 +/- 4.7 (range 37.9-53.3). Mean operative time was 255 +/- 73 minutes (range 150-360). Mean hospital stay was 3 +/- 1 days. Intraoperative complications were absent. CONCLUSION: Preliminary results have been satisfactory, and encourage us to continue with LASGB. PMID- 9730533 TI - Bariatric surgery at the 1st Surgical Department in Prague: history and some technical aspects. AB - BACKGROUND: Obesity has been increasing in the Czech Republic over the last 20 years. In 1983 we were one of the first surgical departments in the country which performed bariatric surgery on a regular basis. METHODS: From 1983 to 1986 we performed vertical banded gastroplasty (VBG). Because of a high rate of various complications arising both from the stomach and the wound, we switched in 1986 to 'less aggressive' nonadjustable gastric banding (GB). In 1993 we performed the first laparoscopic nonadjustable banding, and in 1994 we started laparoscopic placement of adjustable gastric bands. RESULTS: In the group of 52 patients who underwent VBG and were followed-up, acceptable weight loss results (-40.5 kg) were achieved in the 24 months following surgery. The postoperative complications were high; 17.3% gastric staple-line disruption and 15.3% wound complications (incisional hernias, discharge, etc.). Since 1986, we have performed nonadjustable GB in 150 patients and achieved weight loss of -38.4 kg in the 24 months following surgery. There was no change in the wound complication rate, but the complications arising from the stomach and the band decreased to 6.3%. Since June 1993, we have performed 268 procedures laparoscopically, either with nonadjustable bands or, since 1994, with the adjustable bands. The wound complication rate decreased to 0.9%, and one complication (6.6%) was related with the adjustable band. CONCLUSIONS: Because of the high rate of postoperative complications in our experience with VBG, we started GB in 1986. Since then the number of complications arising from the stomach has decreased substantially. With the laparoscopic technique, there was a further decrease in wound healing problems. With the adjustable GB, a significant decrease in the stomach-related complications occurred. Shorter hospital stays were possible with the laparoscopic technique. Long-term weight loss results have not been significantly different among the above mentioned procedures. PMID- 9730534 TI - Biliopancreatic diversion: clinical experience. AB - BACKGROUND: Biliopancreatic diversion (BPD), by ad hoc stomach resection (AHS BPD) has been accepted as an effective surgical treatment for morbid obesity. METHODS: Between 1.1.1992 and 31.7.1996, 59 patients (54 females, five males, mean age 40.3 years, range 23-61 years) underwent AHS-BPD. Mean preoperative body weight was 121.2 kg (range 94-160), with a mean body mass index of 48.6 (range 35 64). Three of these patients were converted from a previous vertical banded gastroplasty to AHS-BPD (one patient with stomach preservation). After at least 36 months follow-up, seven patients underwent abdominal dermolipectomy (five with associated incisional hernia repair, one with thigh dermolipectomy). RESULTS: Mean post-operative hospital stay was 13 days (range 10-30 days). Follow-up is currently in progress in all patients. Excess body weight-loss was 78% in 33 patients with 24 months follow-up, with excellent long-term weight loss maintenance. Protein deficiency was the main specific complication, encountered in two patients (3.4%). Mortality was one patient (1.7%), due to pulmonary embolus. CONCLUSIONS: This clinical experience supports the effectiveness and safety of AHS-BPD, despite some criticism. This procedure appears to be suitable for patients with clinically severe obesity who will poorly tolerate food intake restriction but will accept long-term follow-up. Careful preoperative clinical assessment and selection of patients who will be reliable in long-term follow-up are the keys to success with AHS-BPD, both in terms of weight loss and reduction of specific metabolic complications. PMID- 9730535 TI - Biliopancreatic diversion with transitory gastroplasty preserving duodenal bulb: 3 years experience. AB - BACKGROUND: The authors have performed 521 bariatric surgery operations (319 restrictive procedures and 202 malabsorptive procedures). METHODS: During the last few years we have introduced an evolution of biliopancreatic diversion (BPD): BPD with transitory gastroplasty, preserving the duodenal bulb (53 cases). From a technical point of view, the operation consists of a BPD, coupled with a gastroplasty which is transitory due to the use of a polydioxanone (PDS) band. In the last few cases, instead of a VBG (with PDS band) in order to make the operation completely reversible without any suture on the stomach, we made a gastric pouch by banding with PDS calibrated with the same tube as for the Lap band (20 cc). We maintained completely the duodenal bulb (5 cm from the pylorus), making an end-to-side duodeno-ileal isoperistaltic anastomosis. RESULTS: With this anastomosis, only 2% of patients developed an anastomotic ulcer. With this new procedure, results have been good in terms of weight loss (similar to that of BPD-AHS) and in nutritional complications. No patient has had hypoalbuminemia, diarrhea or halitosis. CONCLUSION: BPD with temporary gastric restriction has provided satisfactory results. PMID- 9730536 TI - Two cases of conversion of vertical ring gastroplasty to adjustable silicone gastric banding. AB - BACKGROUND: Since May 1995, we have used laparoscopic adjustable silicone gastric banding (ASGB) as an alternative to silastic ring vertical gastroplasty (SRVG) to treat morbid obesity. Moreover, it seemed that ASGB was an appropriate procedure to use when SRVG had failed and no alternative procedure could be attempted again, which occurred in two patients. Because of adhesions, the laparoscopic approach was inappropriate in both cases. The size of the pouch and the staple line were not obstacles to ASGB. METHODS: Case 1 was a 53-year-old woman of 111 kg (BMI = 46) who had SRVG in July 1994. One year later, she had a 54 kg weight loss, but had continuing food intolerance, although malfunction of the pouch or ring could not be found. A removal of the ring was performed in October 1995, and a 10-cm diameter silicone band placed. The band was not inflated until she had regained weight 5 months later. Case 2 was a 33-year-old woman of 100 kg (BMI = 40) who had SRVG in March 1994. Weight loss was 45 kg 18 months later; then she gained weight. Endoscopy and barium swallow showed both staple-line disruption and band erosion. Removal of the ring was performed in March 1996, and a 9.75-cm diameter silicone band placed, and inflated at the same time with 2 cc saline. RESULTS: Both patients are doing well. CONCLUSIONS: ASGB appeared to be the best alternative when revising an SRVG in cases where a new stapling or the placement of a new ring could have had consequences more serious than the primary complications. PMID- 9730537 TI - International Federation for the Surgery of Obesity. Statement on morbid obesity and its treatment. PMID- 9730538 TI - International Federation for the Surgery of Obesity. Statement on patient selection for bariatric surgery. PMID- 9730539 TI - Lipoprotein A levels after intestinal bypass operation for morbid obesity. AB - BACKGROUND: Serum Lipoprotein A [Lp(a)] is considered an independent risk factor for cardiovascular disease. Reduction of other risk factors such as serum triglycerides and serum cholesterol is seen after weight reduction but it has been difficult to demonstrate a similar reduction of Lp(a). In this study Lp(a) is studied following weight reduction after intestinal bypass surgery for obesity. METHODS: A cross-sectional study was performed in which Lp(a) levels were compared between patients operated with intestinal bypass surgery for obesity (bilio-intestinal bypass) and a closely matched control group. The controls were chosen so as to correspond to the operated patients' preoperative body mass indices. RESULTS: The operated group had reduced their body mass index from a mean of 42 kg/m2 to a mean of 29 kg/m2 and had a significantly lower mean serum Lp(a) level (113 mg/l, SEM 34) than controls (207 mg/l, SEM 37), p < 0.05. CONCLUSION: Lowered Lp(a) levels are correlated to substantial weight loss following intestinal bypass surgery for obesity. PMID- 9730540 TI - Long-term results of vertical banded gastroplasty: Marlex versus Dacron banding. AB - BACKGROUND: The VBG was originally performed with a Marlex band and characterized by a satisfactory weight loss and low morbidity. The effect of the material used for the banding procedure (Marlex vs Dacron) in vertical banded gastroplasty (VBG) is evaluated. METHODS: In 49 consecutive obese patients treated with a VBG, a Marlex band was used in 17 patients and a Dacron band in 32 patients. Data were analyzed retrospectively with regard to the type of band, weight loss and complications. RESULTS: A significant difference was found in the percentage excess weight 5 years postoperatively in favor of the Dacron group (59.2% vs 39.2%; p < 0.05) because of more band-related complications in the Marlex group. The difference in percentage excess weight disappeared 8 years postoperatively (43.3% vs 46.8%), due to the renewed weight loss of the Marlex group following reoperation. CONCLUSION: The Dacron band is superior to the Marlex band in VBG because sustained weight loss is satisfactory and morbidity is low. PMID- 9730541 TI - Staple disruption in vertical banded gastroplasty. AB - BACKGROUND: In earlier studies of vertical banded gastroplasty (VBG), staple-line disruptions (SD) were only reported in a few per cent or less. Two recent studies have shown a significantly higher frequency of SD. To find the true frequency of SD, all patients must be examined regardless of weight outcome. METHODS: 91 consecutive patients were examined by a standardized radiological method at different postoperative intervals ranging from 6 to 48 months. RESULTS: 41 out of 91 patients developed SD. The average diameter of the disruptions was 6 mm (range 2-16 mm). During the first 2 years of follow-up, when at least 31% of the patients had developed SD, there was no significant difference in weight loss between the group with SD and the group without SD. CONCLUSION: SD is an inherent problem of VBG which has been underestimated for a long period of time. An SD frequency of 45% or more within the first few years is not acceptable and changes in the VBG technique must be considered. PMID- 9730542 TI - Weight loss following transected gastric bypass with proximal Roux-en-Y. AB - BACKGROUND: Cross-sectional analysis was performed to study the trend in weight loss following surgery. METHODS: Among 194 patients involved in the study during the period 1990-1995, 86.6% were female and 87.1% were white. Median values for the initial cohort of 194 patients were as follows: age 32.5 years; height 163.8cm; preoperative weight 122.81 kg. Of these patients 67%, 30%, and 3% of the patients were categorized as morbidly obese, super obese, and obese respectively. They all underwent gastric bypass surgery. RESULTS: Follow-up rates were 67.9% at 1 year, 55.8% at 2 years, 62.8% at 3 years, and 70.0% at 4 years postoperation. Median BMI was reduced from 45.18 to 28.40, and median percentage loss of excess weight was 68.5% after 1 year. After 2, 3, and 4 years, median BMI values were 27.69, 28.63, and 29.40, and median percentages of excess weight loss were 71.18%, 69.28%, and 57.49%, respectively. Although the analysis at 4 years indicates that some patients experience slight weight gain, the increase was not significant and further analysis will be performed when more data points are available. CONCLUSION: Postoperative weight loss has been satisfactory. PMID- 9730543 TI - Reducing early technical complications in gastric bypass surgery. AB - BACKGROUND: The incidence of complications following gastric bypass surgery has decreased markedly over the last 30 years; nevertheless, significant morbidity and mortality is still associated with this procedure. Much of the improved risk of this technique can be attributed to the numerous modifications that have taken place in its evolution. METHODS: We compared our series of 640 primary cases of vertical banded gastroplasty-Roux-en-Y gastric bypass (VBG-RGB), a form of gastric bypass, with gastric bypass series reported in the literature from 1966 to 1996. Incidences considered were those of subphrenic abscess, gastrointestinal leaks, obstruction of the excluded segment of gastrointestinal tract, splenectomy and death. RESULTS: The overall trend during the last 30 years has been a reduction in the rate of major complications. In our series, we had one major complication, a subphrenic abscess. This compares favorably with the incidence of major complications reported in the literature. CONCLUSIONS: The gastric bypass is a significantly safer operation today than three decades ago. We believe that the relatively low complication rate of VBG-RGB results from: (1) the anatomic location of the gastric pouch; (2) the type of stapling device used in its construction; (3) a pouch outlet restricted by a prosthetic band rather than a narrow anastomosis; and (4) the construction of a retrocolic, retrogastric Roux en-Y gastrojejunal anastomosis. PMID- 9730544 TI - Massively bleeding gastric pouch ulcer after silastic ring vertical gastroplasty successfully treated endoscopically: a report of two cases. AB - Two patients who developed massive bleeding from a gastric pouch ulcer are described. This rare complication occurred during the early postoperative course after silicone ring vertical gastroplasty (SRVG). In both cases the bleeding stopped after the ulcers were injected with epinephrine and alcohol. Both ulcers healed after 1 month of treatment with omeprazole (Losec). The probable etiology of this rare complication is discussed. PMID- 9730546 TI - Body composition and source of weight loss after bariatric surgery. AB - BACKGROUND: Little is known about the composition and source of weight loss after bariatric surgery for morbid obesity. PURPOSE: This study was undertaken to determine changes in weight, body mass index (BMI), lean body weight (LBW), fat weight (FW) and left ventricular cardiac mass (LVM) following vertical banded gastroplasty (VBG). METHODS: After VBG for morbid obesity, 26 women and four men (mean age = 39.1 years) were weighed and had body composition analysis undertaken at intervals. Thirteen patients underwent echocardiography preoperatively and 1 year postoperatively to determine change in LVM and LVM index. RESULTS: Over 12 months there was significant weight loss for all weight parameters examined (p < 0.05). Fat weight loss was most significant; total weight loss and reduction of BMI were significant but less so than fat loss (Wilcoxon's signed ranks test). LBW loss had the smallest contribution to weight loss (p < 0.0001). There was a significant loss of LVM and posterior cardiac wall thickness (p < 0.05). CONCLUSIONS: VBG can lead to loss of lean body weight and left ventricular mass, and more dramatically, fat weight, body weight, and BMI. Cardiac mass and lean body mass are preferentially conserved relative to body fat with weight loss after VBG. PMID- 9730547 TI - A decade of change in obesity surgery. National Bariatric Surgery Registry (NBSR) Contributors. AB - BACKGROUND: The International (formerly National) Bariatric Surgery Registry began collecting data in January 1986. The aim of this study was to examine changes in the practice of surgical treatment of severe obesity that occurred during the decade of 1986 through 1995, as observed in the IBSR data. METHODS: All data submitted to the IBSR during the decade were transferred to the IBM mainframe computer for analysis. Characteristics of operative type populations were compared over time using analysis of variance (ANOVA) for age, body mass index (BMI), operative weight and Chi-square (chi2) test for gender. RESULTS: There has been a steady increase over the decade in mean patient weight. The operations used have changed from predominantly 'simple' operations to more frequent use of 'complex' operations. Within the categories of 'simple' and 'complex', an increase in the variety of operations occurred. As a group, patients with 'simple' operations have been heavier, more often male and public pay patients than those who have undergone 'complex' operations. One year weight loss was greater for Roux-en-Y gastric bypass (RGB) than vertical banded gastroplasty (VBG), but follow-up rates were too low to study the relative merits of the operations used. The reported incidence of operative mortality and serious complications (leak with peritonitis, abscess and pulmonary embolism) remained low. CONCLUSIONS: These observations and their implications can be summarized in three statements which relate to action for improved patient care in the beginning of the new century: (1) increasing weight of candidates for surgical treatment during this decade indicates the need for earlier use of operative treatment before irreversible complications of obesity can develop; (2) low risk of obesity surgery, decreasing postoperative hospital stay, and early weight control support the continued and increased use of surgical treatment; (3) continued widespread use of both 'simple' and 'complex' operations with increased modifications of standard RGB and VBG procedures emphasizes the need for standardized long-term data and analyses regarding both weight control and postoperative side-effects. PMID- 9730548 TI - Gastric bypass for morbid obesity: a standard surgical technique by consensus. AB - BACKGROUND: The gastric bypass operation has evolved since 1966 when it was first introduced. The purpose of this study was to determine the present state of gastric bypass by consensus among the members of the American Society for Bariatric Surgery (ASBS). METHOD: A questionnaire was sent to all members of the ASBS. Forty-three percent responded reporting over 41,200 cases. RESULTS: Results were analyzed by using chi2 tests with a null hypothesis. Surgeons agreed on several technical aspects, preferring a vertical to a horizontal stapleline; estimating, rather than measuring, the pouch volume at an average of 22 cc. Few surgeons divide the short gastric vessels, and only 25% of surgeons polled use a restrictive ring or band proximal to the gastroenterostomy. Most surgeons calibrate the gastroenterostomy, reporting a preferred average diameter of 12.3cm. There was no consensus regarding forming the gastroenterostomy, 58% preferring hand-sewn and 42% stapled anastomoses. There was no consensus regarding dividing the gastric pouch from the bypassed stomach: CONCLUSION: The preferred gastric bypass is vertical, with the pouch estimated at 20-25 cc, and the gastroenterostomy calibrated at 12 mm diameter. The short gastric vessels need not be divided, and restrictive bands or rings are not preferred. This technique of gastric bypass should be used as the control procedure when modifications are tested in future trials. Randomized prospective studies are suggested to probe the benefits of division of the stomach pouch from the bypassed stomach. PMID- 9730549 TI - Swedish adjustable gastric banding: a preliminary experience. AB - BACKGROUND: The authors have been performing bariatric surgery for 15 years; since February 1992 they have carried out laparoscopic gastric banding (LGB) with a silastic band. Good experience with the LGB combined with everyday laparoscopic activity in their institution persuaded them to try laparoscopic placement of the Swedish adjustable gastric band (SAGB). METHODS: The surgical procedure is the same as that for laparoscopic gastric banding except for the use of a 15 mm trocar that is required to introduce the band, and the need to place a port that is connected to the band via a wide loop tube; the port is place subcutaneously and is supported by the lower part of the sternum. The authors do not use any abdominal drain nor a naso-gastric tube. At surgery the band is left empty, and filling is usually not started until 4 weeks after surgery. The patient is immediately mobilized and begins a liquid diet the evening after the operation. The patients are usually discharged from hospital on the first postoperative day. RESULTS: Over 8 months, 24 patients underwent SAGB, with mean BMI 44.69 and mean operating time 45 minutes (range 30-75). No early complications occurred. Preliminary results in this small series show BMIs of 38.65 and 34.60 at 3 and 6 months postoperation. CONCLUSION: SAGB appears for be a good method for obesity surgery. It is easy to perform and is associated with a low operative risk. Provided that the band is put in the right place, weight loss can be adjusted to patient comfort. PMID- 9730550 TI - Stapler division of the omentum and small bowel mesentery in morbidly obese patients undergoing gastric bypass surgery. AB - BACKGROUND: Roux-en-Y gastric bypass (RYGB) procedures can be technically demanding because of anatomical factors including fat distribution, organ fixation, and wound depth. We developed a technique using the Multifire Endo GIA 60 2.5 disposable surgical stapler which allows greater mobilization, less blood loss, and decreased operating time. METHODS: A disposable stapler designed for laparoscopic surgery was used to transect the gastro-colic omentum and small bowel mesentery in 67 morbidly obese patients undergoing RYGB. Generally, five to six stapler cartridges were needed for the transections. RESULTS: Stapler division of the gastro-colic omentum and small bowel mesentery decreased operating time by an average of 25 minutes. CONCLUSIONS: Applying principles of laparoscopic surgery to RYGB resulted in more efficient mobilization of the stomach and the small bowel mesentery, as well as decreased blood loss and operating time. PMID- 9730551 TI - Incarcerated Spigelian hernia in morbidly obese patients: the role of intraoperative ultrasonography for hernia localization. AB - BACKGROUND: Spigelian hernias are uncommon and their diagnosis can be difficult. Ultrasonography is, as a rule helpful in making the diagnosis, but extensive exploration is sometimes needed to locate the defect. METHODS: Two patients are described in whom the diagnosis was made preoperatively by ultrasonography, but intraoperative location of the hernias proved extremely difficult because of the patients' obesity. RESULTS: In the first patient, the hernia was located by means of intraoperatively performed ultrasonography. In the second patient, ultrasonography was combined with intraoperatively insufflated pneumoperitoneum and this proved successful in identifying the position of the sac. CONCLUSION: Intraoperative ultrasonography is a valid option for accurate localization of Spigelian hernias, especially in obese patients; extensive intraoperative dissection, distortion of tissue planes, and associated morbidity risks may be avoided. PMID- 9730552 TI - An approach to the treatment of recurring polyarthralgia after jejunoileal bypass. AB - Polyarthralgia after jejunoileal bypass (JI) is a well-documented complication. In the past, this was treated by antibiotic therapy, but definitive therapy uncommonly necessitated surgical reversal of the JI bypass. This case report discusses the etiology of arthralgias and presents a technique for its treatment without bypass reversal. PMID- 9730554 TI - Papers related to obesity surgery for 1996. PMID- 9730556 TI - Effect of L-carnitine and palmitoyl-L-carnitine on erythroid colony formation in fetal mouse liver cell culture. AB - The administration of L-carnitine to patients undergoing hemodialysis increases hematocrit and improves the response to recombinant human erythropoietin (rhEPO). This suggests a contribution by carnitine to erythrocyte membrane function or erythropoiesis. We investigated the effect of L-carnitine and palmitoyl-L carnitine on erythropoiesis by assessing erythroid colony formation in in vitro fetal mouse liver cell cultures. L-Carnitine or palmitoyl-L-carnitine was added with rhEPO to fetal mouse liver cell cultures. Doses of L-carnitine of up to 200 micromol/l to the culture had no effect on colony formation. In contrast, the addition of above 12.5 micromol/l palmitoyl-L-carnitine into the culture increased colony formation significantly. These results suggest that long-chain acyl carnitine may have an effect on erythropoiesis. PMID- 9730557 TI - Metabolic factors in the renal response to amino acid infusion. AB - In order to investigate the renal effects of amino acids (AA) with different metabolic fate, we compared the changes in glomerular and tubular function, nitrogen metabolism and glucoregulatory hormones in 7 volunteers during two infusions, one of a complete solution of amino acids (MIX-AA), which included five AA electively metabolized at the splanchnic level, and the other of a solution containing only essential AA (EAA), which escape splanchnic metabolism. MIX-AA increased GFR and RPF (from 104 +/- 6 to 122 +/- 13 and from 488 +/- 46 to 572 +/- 34 ml/min/1.73 m2), stimulated splanchnic metabolism as demonstrated by rises in urinary urea excretion (from 20.7 +/- 2 to 30.6 +/- 7.5 mg/min/1.73 m2) and the plasma glucagon/insulin ratio (from 21.4 +/- 13 to 26.7 +/- 15), and caused increases in fractional excretion of AA, FeNa and free-water clearance. During MIX-AA infusion significant correlations were observed between the individual values of GFR and the urea excretion rate (r = 0.66), and between GFR modifications (DeltaGFR) and the plasma glucagon/plasma insulin ratio (r = 0.40). No change in renal function, urea excretion and the glucagon/insulin ratio was observed with EAA. An intermediate splanchnic step (increased plasma glucagon/insulin ratio and ureagenesis) seems necessary in the pathway leading to the nonessential AA-induced rise in GFR; this might stimulate an ultimate intrarenal pathway (possibly involving the diluting segment) via a still undefined mechanism. PMID- 9730558 TI - Plasma homocysteine, vitamin B6, vitamin B12 and folic acid in end-stage renal disease during low-dose supplementation with folic acid. AB - In order to see whether conventional low-dose folic acid supplement along with vitamin B6 and B12 reduces hyperhomocysteinemia in patients with ESRD, we compared the levels of homocysteine, vitamin B6, B12 and folic acid among 3 groups of patients: 44 ESRD patients on hemodialysis with replacement of folic acid, vitamin B6, and B12 (dialysis group); 27 chronic renal failure patients without vitamin replacement (predialysis group); and 17 hypertensive patients without vitamin replacement (control group). Mean plasma total homocysteine concentration was higher in the dialysis (15.5 +/- 6.6 micromol/l) and the predialysis groups (15.7 +/- 4.2 micromol/l) than in the control group (6.2 +/- 1.5 micromol/l) (p < 0.001). However, there was no difference in homocysteine concentrations between the dialysis and predialysis groups. In the control and predialysis groups, the homocysteine concentration showed a reverse correlation with the concentrations of folic acid (r = 0.584, p = 0. 014 for the control group; r = 0.431, p = 0.247 for the predialysis group) and vitamin B12 (r = 0.485, p = 0.049 for the control group; r = -0.562, p = 0.023 for the predialysis group) but not with vitamin B6. In conclusion, plasma folic acid concentrations were 3-4 times higher in the dialysis than in the predialysis group. But these levels of folic acid are not enough to reduce hyperhomocysteinemia in ESRD. PMID- 9730559 TI - Clinical course of cytomegalovirus (CMV) viremia with and without ganciclovir treatment in CMV-seropositive kidney transplant recipients. Longitudinal follow up of CMV pp65 antigenemia assay. AB - This study was designed to evaluate the longitudinal history of cytomegalovirus (CMV) infection and to test the capacity of ganciclovir as effective therapy in CMV-seropositive renal transplant recipients. The CMV viremia was detected with CMV pp65 antigenemia assay in 153 renal transplants. The recipients were classified as having low-grade and high-grade CMV infections according to the severity of CMV infection. The recipients with low-grade CMV infections were observed without ganciclovir treatment, and the recipients with high-grade CMV infection were randomly assigned to ganciclovir-treated and untreated groups. The clinical course between low-grade and high-grade CMV infections was evaluated. All recipients with low-grade CMV infection (n = 62) showed spontaneous remission regardless of immunosuppresants. In high-grade CMV infection (n = 31), the ciclosporin A treated group (n = 11) showed no evidence of CMV disease, and the methylprednisolone-treated group (n = 8) showed CMV disease in 1 (25%) of 4 ganciclovir-untreated recipients. In the OKT3 group (n = 12), symptomatic CMV infection was observed in 6 (100%) ganciclovir-untreated recipients contrary to no CMV disease in the ganciclovir-treated group (p < 0.05). In conclusion, the CMV antigenemia assay is effective in monitoring CMV viremia, and ganciclovir treatment should be done during early CMV viremia in OKT3-treated recipients. PMID- 9730560 TI - The hemodialysis access: preferences and concerns of patients, dialysis nurses and technicians, and physicians. AB - One hundred twenty-eight hemodialysis patients and 64 medical personnel consisting of dialysis nurses and technicians, hemodialysis access surgeons and nephrologists were surveyed about their preferences and concerns in regard to the hemodialysis vascular access. The access preferred by physicians was the A-V fistula in the lower arm. In contrast, the access preferred by dialysis nurses and technicians was the polytetrafluoroethylene (PTFE) graft in the lower arm. Patients desired a superficial access in the forearm which was easy to cannulate, had minimal effect on their appearance, provided quick hemostasis after dialysis and enabled arm comfort during dialysis. Physicians felt the most significant concerns about the access were thrombosis and infection. Nurses and technicians ranked difficult cannulation and insufficient access blood flows that prohibited dialysis adequacy as their major problems. For patients the most common problem was pain during needle insertion. This survey concluded that the A-V fistula remains the access of choice. However, appropriate maturation of the fistula must occur before needle insertion is attempted. An immature fistula is difficult to cannulate, has fragile veins resulting in blood leakage around the needle infiltrating the subcutaneous tissues and has inadequate blood flows for successful dialysis. Patients who are introduced to dialysis with inadequate access function or access failure from either an A-V fistula or a PTFE graft have increased morbidity, inadequate dialysis and enhanced anxiety about dialysis treatments. To increase the success and acceptance of A-V fistulas in hemodialysis patients it is incumbent upon the nephrologist to protect the future access arm from damage to the vasculature and to allow for fistula maturation before cannulation. Surgical protocols must improve the appropriate selection of a fistula or PTFE graft for various age groups and disease categories. Better patient preparation and selection of the proper access type for each patient will enhance early access function and subsequent access survival. PMID- 9730561 TI - Low molecular weight heparin reduces triglyceride, VLDL and cholesterol/HDL levels in hyperlipidemic diabetic patients on hemodialysis. AB - BACKGROUND: Low molecular weight heparin (LMWH) provides a safe and effective alternative for hemodialysis anticoagulation. While unfractionated (UF) heparin has been implicated in hyperlipidemia, the effect of LMWH on the lipid profile in nondiabetic patients is controversial in chronic hemodialysis. The effect of LMWH in diabetic patients, a high risk group of hyperlipidemia, has not been studied. METHOD: LMWH was tested for its safety and efficacy in 10 nondiabetic Taiwanese patients. To evaluate influence of lipid profile, a crossover study was carried out in 10 type II diabetic patients with poor blood sugar control associated with high triglyceride (430.4 +/- 101.1 mg/dl) and total cholesterol levels (219.2 +/- 12.7 mg/dl) using UF heparin for more than 1 year. These patients were subjected to Fraxiparine, an LMWH, for 6 months and then switched back to UF heparin for another 6 months. Lipid profiles were measured every 2 months without prescribing lipid-lowering agents and the blood sugar was maintained at stationary levels. RESULTS: LMWH is safe and effective in Taiwanese patients as a single bolus injection and maintains a 9.4% higher platelet count immediate postdialysis compared to UF heparin. With high HbA1c levels (9.6 +/- 0.6%), mean triglyceride and VLDL levels started to decrease at the 4th month after LMWH treatment and reached a 34% reduction in triglyceride, a 26.2% reduction in VLDL, and a 19% reduction of total cholesterol/HDL ratio at the 6th month. Increments of triglyceride levels were found at the 6th month after a switch back to UF heparin. The levels of total cholesterol, LDL-cholesterol, HDL-cholesterol, apolipoprotein A-1 and B remained unchanged. CONCLUSION: LMWH may be beneficial to lipid control in hyperlipidemic diabetic patients on hemodialysis. PMID- 9730562 TI - Sympathetic activity and blood pressure pattern in autosomal dominant polycystic kidney disease hypertensives. AB - To study the potential role of sympathetic activity in the pathogenesis of arterial hypertension associated with autosomal dominant polycystic kidney disease (ADPKD) and to analyze its relationship with 24-hour blood pressure pattern, plasma catecholamines and 24-hour ambulatory blood pressure monitoring were evaluated in 30 ADPKD hypertensive patients (of which 17 without and 13 with renal failure) and in 50 essential hypertensives. The groups were matched for sex, body mass index, known duration of hypertension, and clinic blood pressure. Plasma catecholamines, determined in resting position, were higher in ADPKD patients without renal failure than in essential hypertensives. Nighttime diastolic blood pressure was higher and the percentage day-night difference in mean blood pressure was lower in hypertensives with ADPKD compared to patients with essential hypertension. Blood pressure was significantly correlated with plasma noradrenaline in ADPKD patients, independently of renal function. No significant differences were observed between ADPKD patients with and without renal failure, with respect to plasma catecholamines, 24-hour daytime and nighttime ambulatory blood pressures and the percentage day-night difference in mean blood pressure. PMID- 9730563 TI - Study of effect of optimization of dialysis and protein intake on neuromuscular function in patients under maintenance hemodialysis treatment. AB - This study was carried out on 22 patients on maintenance hemodialysis. Among them, 20 patients were males and 2 were females, their age ranged from 12 to 50 years. Initially, the patients were assessed clinically and by laboratory investigations and their dialysis was assessed by studying their urea kinetic modeling following the nomographic approach for calculating their Kt/V values. Their nutrition was assessed by measuring skin folds, midarm circumference, laboratory parameters and by calculating the normalized protein catabolic rate (nPCR). Also their neuromuscular functions were assessed by clinical examination and neurophysiologic study. Dialysis dose was readjusted to achieve a target Kt/V value of 1.3 for patients on 3 times weekly dialysis and 1.6 for patients on twice weekly dialysis. Also, their nutrition was reviewed to achieve nPCR 1.2 g/kg/day and caloric intake 30-40 kcal/kg/day through diet manipulation and support. The patients were assessed finally after 3 months on targeted dialysis and nutrition by thorough clinical, laboratory and neuromuscular assessment. Analysis of neurophysiologic data showed significant improvement in electromyography. Furthermore, fatigue test showed significant (p = 0.002) decreases in muscle fatigue after optimization of dialysis dose and patients' nutrition. From this study, we may conclude that in dialysis patients, even when asymptomatic and clinically stable, neurologic deficits do exist and using area kinetic modeling to improve dialysis and patients' nutrition is valuable in improving their neuromuscular functions. PMID- 9730564 TI - Effect of intravenous calcitriol on cardiac systolic and diastolic function in patients on hemodialysis. AB - The systolic and diastolic function of the heart of hemodialysis (HD) patients and the effect of intravenous vitamin D therapy on cardiac function was studied by Doppler and digitized M-mode echocardiography in 10 HD patients before and after 3-4.5 months of calcitriol therapy. Calcitriol was administered intravenously 1-3 times a week at a dose of 1-2 microg after the dialysis sessions. Ten age- and sex-matched healthy controls were also examined echocardiographically. Before calcitriol therapy cardiac wall thicknesses (interventricular septum, posterior wall) and left ventricle (LV) dimensions (end diastolic, end systolic) were greater, and LV diastolic (peak late diastolic velocity, peak early diastolic velocity/peak late diastolic velocity ratio, isovolumic relaxation time) and systolic (fractional shortening) function was impaired in HD patients as compared to controls. The LV posterior wall thickness was related to plasma parathyroid hormone (PTH; r = 0. 70, p = 0.01) in the patients. Calcitriol therapy raised serum ionized Ca from 1.23+/-0.04 to 1.33 +/- 0.04 mmol/l and reduced PTH from 41.1+/-10.7 to 34.2+/-11.7 pmol/l (29+/-11%). Calcitriol therapy did not cause any significant changes in cardiac function in the whole patient group. However, in a subgroup of 5 patients with severe but controllable hyperparathyroidism (PTH >3 times upper normal margin) the LV dimensions and systolic function improved (LV end systolic dimension from 39.0 +/ 4.0 to 31.3 +/- 2.9 mm, p = 0. 03; LV end diastolic dimension from 57.7 +/- 3.1 to 53.4 +/- 3.0 mm, p = 0.06; fractional shortening from 33 +/- 4 to 42 +/- 3%, p = 0. 03). The diastolic indices improved also, but not significantly. In conclusion, left ventricle hypertrophy and systolic and diastolic dysfunction was observed in HD patients. Intravenous calcitriol therapy improved cardiac function in patients with severe secondary hyperparathyroidism. PMID- 9730565 TI - Plasma concentration of brain natriuretic peptide as an indicator of cardiac ventricular function in patients on hemodialysis. AB - The plasma concentration of human brain natriuretic peptide (BNP) was measured by immunoradiometric assay in patients on maintenance hemodialysis (HD) to assess the possible relationship between the plasma levels of this peptide and cardiac ventricular function, as judged by M-mode echocardiography. The plasma BNP levels in the pre-HD state were significantly higher (688.5 +/- 154.5 pg/ml) than those of healthy subjects (<40 pg/ml). In addition, the plasma BNP levels were slightly decreased during HD (post-HD, 617.3 +/- 157.1 pg/ml). There was no correlation between the plasma levels of BNP and body weight changes during HD. The mean plasma BNP level was significantly higher in the group of patients with a low left ventricular ejection fraction (EF < 60%) than in the group with a normal EF. In the patients as a whole, there was an inverse correlation between plasma BNP levels and EF. Moreover, a positive correlation was found between plasma BNP levels and left ventricular mass index (r = 0.57, p < 0.05). These results suggest that plasma BNP levels increase in response to chronic stimulation in accordance with increased cardiac load, and that they may be a possible indicator of reduced ventricular function in HD patients. PMID- 9730567 TI - A case of IgA nephropathy in three sisters with thin basement membrane disease. AB - IgA nephropathy associated with thin basement membrane disease is reported in a 9 year-old female. The diagnosis of IgA nephropathy was made by means of an immunofluorescence investigation, which showed generalized diffuse mesangial deposits. Thin basement membrane disease was identified by electron-microscopic investigations, which disclosed thinning of the basement membrane of several capillary loops and prominence of the lamina densa. Her father, elder sister and younger sister were also found to have hematuria and her sisters were diagnosed as having thin basement membrane disease by renal biopsy. Patients with IgA nephropathy have focal thinning of the glomerular basement membrane, but we consider that urinalysis of the family needs to be done for the diagnosis of familial thin basement membrane disease, when diffuse thinning of the glomerular basement membrane is detected in such patients. PMID- 9730566 TI - Effect of L-carnitine supplementation on lipid parameters in hemodialysis patients. AB - It has been reported that cumulative carnitine losses through dialysis membranes may worsen hyperlipidemia during long-term hemodialysis. However, carnitine supplementation has not shown a consistent beneficial response. We undertook the present study to determine if there is any hypolipidemic effect of L-carnitine on Greek dialysis patients in concert with the dialysate buffer composition (acetate or bicarbonate). A total of 28 patients (16 male, 12 female), mean age 43 years (range 21-61), with end-stage renal disease on maintenance hemodialysis for a mean period of 25 months (range 7-84) were studied. The dialysis schedule was 4 h, 3 times/week using cuprophane hollow-fiber dialyzers and acetate (n = 14) or bicarbonate (n = 14) dialysate. In all patients L-carnitine (5 mg/kg body weight) was infused intravenously 3 times/week at the end of each hemodialysis session. Blood samples for carnitine and lipid determinations were obtained before treatment, and 3 and 6 months following treatment. Even though L-carnitine did not modify most of the serum lipid levels, a significant decrease in serum triglycerides was evident in the whole group of patients (from 225 +/- 76 to 201 +/- 75 mg/dl, p = 0.03). Furthermore, L-carnitine could decrease serum triglycerides only in hypertriglyceridemic patients (from 260 +/- 64 to 226 +/- 82 mg/dl, p < 0.05). L-Carnitine resulted in a reduction of serum triglycerides in both patients on bicarbonate and on acetate dialysis, while there were no significant differences in the changes of lipid parameters after L-carnitine between the two groups of hemodialysis patients. We conclude that relatively low doses of L-carnitine supplementation could contribute to the management of some hypertriglyceridemic hemodialysis patients. PMID- 9730568 TI - IgA nephropathy in psoriasis. AB - IgA nephropathy (IGAN) was first described by Berger and Hinglais in 1968. It is now the most common glomerular disease worldwide. IGAN has been associated with several diseases. Its association with psoriasis has been rarely described. We report a case of IGAN with crescentic changes, associated with psoriasis vulgaris. We review the literature on the association of IGAN with psoriasis and discuss the likely pathogenetic linkage. PMID- 9730569 TI - Isoniazid-induced crescentic glomerulonephritis in a child with a positive tuberculin skin test. AB - Acute nonoliguric renal failure developed in a 13-year-old girl, 1 month after the institution of isoniazid therapy because of a positive tuberculin test at school screening. A renal biopsy demonstrated severe crescentic glomerulonephritis with focal interstitial changes. Discontinuation of isoniazid and a short course of steroids and cyclophosphamide therapy were followed by complete recovery. Whereas isoniazid has been shown to induce a lupus-like syndrome and antihistone antinuclear antibodies, our patient displayed none of the clinical or immunological features that are characteristic of drug-induced lupus. Furthermore, none of the identifiable causes for crescentic glomerulonephritis was evident in this girl. To the best of our knowledge this is the first report suggesting a possible association of crescentic glomerulonephritis to isoniazid treatment. PMID- 9730570 TI - Recovery of minimal change nephrotic syndrome and acute renal failure in a patient withRenal cell carcinoma. AB - We describe a 78-year-old patient with nephrotic syndrome due to minimal-change glomerulopathy, associated with a renal adenocarcinoma. Oliguric acute renal failure requiring hemodialysis was also observed. Surgical removal of the tumor and corticosteroid therapy resulted in resolution of the nephrotic state and improvement of the renal function. Nephrotic syndrome is an unusual complication of renal cell carcinomas, and the association of minimal-change glomerulopathy (MCG) and solid tumors is particularly uncommon. In spite of this, MCG should be considered in the nephropathies causing nephrotic syndrome and acute renal failure in patients with renal malignancies. PMID- 9730571 TI - Pseudo-normal osmolal and anion gaps following simultaneous ethanol and methanol ingestion. AB - Methanol, ethylene glycol, and isopropyl alcohol are associated with acute intoxication. The diagnosis is dependent upon high anion-gap metabolic acidosis, and an osmolal gap between the calculated and the measured osmolality. Normal anion gap has been reported in some cases of concomitant methanol and ethanol ingestion, where the high serum levels of ethanol inhibited the metabolism of methanol by alcohol dehydrogenase. The osmolal gap in these cases was higher than expected for methanol, and served as a constant marker for a metabolic derangement. Herewith, we present a patient who presented with normal osmolal and anion gaps 36 h after ethanol and methanol ingestion, yet progressively developing ocular toxicity. Normal anion and osmolal gaps should not rule out earlier methanol poisoning. PMID- 9730572 TI - Appearance of nephrotic syndrome following interferon-alpha therapy in a patient with hepatitis B virus and hepatitis C virus coinfection. AB - A 59-year-old woman with hepatitis B surface antigen (HBsAg) and hepatitis C viral (HCV) antibody presented with proteinuria and hematuria. The patient was treated with interferon-alpha (INF-alpha) because plasma aminotransferase levels had been elevated and a liver biopsy had showed chronic active hepatitis. Her urinary protein excretion decreased as liver function normalized and her serum HCV-RNA was negative during treatment. Eleven weeks after completion of INF-alpha treatment, she suddenly presented with nephrotic-range proteinuria, although an improvement in the hepatic function was maintained. Renal pathologic findings were consistent with membranous glomerulonephritis (MGN), and HBsAg was detected in the glomeruli but not HCV. After treatment with prednisolone, her 24-hour protein excretion was below 0.7 g/day. To our knowledge this is the first report on hepatitis B virus MGN with nephrotic syndrome following IFN-alpha therapy for HCV. This suggests that treatment with INF-alpha might affect the immune processes and may be associated with the pathogenic mechanism in this patient. PMID- 9730573 TI - Acute oliguric renal failure associated with unilateral renal embolism: a successful treatment with iloprost. AB - A 66-year-old woman presented with acute-onset rapid atrial fibrillation and right upper quadrant pain which had appeared 24 h prior to admission. The patient also manifested acute oliguric renal failure (serum creatinine 6.9 mg/dl). Selective renal angiography revealed total occlusion of the right renal artery with normal visualization of the left kidney vasculature. The patient was treated with intra-arterial urokinase and intravenous heparin, with no response. Intravenous administration of the prostacyclin analogue, iloprost, resulted in rapid resolution (within hours) of the oliguric acute renal failure, in spite of the continuing presence of a nonfunctioning right kidney. We conclude that the etiology of the acute renal insult in this patient is probably related to unilateral renal arterial embolization accompanied by arterial spasm of the unaffected kidney. The contralateral vasospasm can be reversed by iloprost, which then leads to a rapid recovery from acute renal failure. We are unaware of prior reports documenting the beneficial effect of iloprost in a clinical setting as described here. PMID- 9730574 TI - Use of recombinant erythropoietin in a pregnant renal transplant recipient. AB - The use of recombinant human erythropoietin (rHuEPO) has been extensively studied among patients with anemia of chronic renal failure. Less well described is its safe use during pregnancy. We report a case of a 17-year-old gravid renal transplant patient with underlying renal insufficiency who became severely anemic and responded to rHuEPO. As the number of renal transplant recipients who become pregnant increases, the need for correction of anemia of chronic renal failure during pregnancy may rise as well. Our case illustrates the use of rHuEPO in this situation. PMID- 9730575 TI - Peritoneal dialysis-associated peritonitis caused by Alcaligenes xylosoxidans. AB - Despite significant progress to decrease its incidence, peritonitis remains the main source of morbidity and treatment failure in patients on continuous ambulatory peritoneal dialysis (CAPD). The majority of cases of peritonitis result from infection with aerobic gram-positive (Staphylococcus epidermidis and Staphylococcus aureus), or gram-negative organisms. Less common organisms that are also reported include anaerobic bacteria, fungi, and mycobacteria, which collectively account for less than 10% of isolates cultured. We report a case of peritoneal dialysis-associated peritonitis, and review the literature on peritonitis caused by Alcaligenes species. Alcaligenes xylosoxidans is a nonfermenting gram-negative rod and opportunistic pathogen that is motile with peritrichous flagella. The clinical features and microbiological data of our case, as well as the other previously reported cases of peritonitis caused by Alcaligenes species show no particular pattern of peritoneal dialysate cell count. However, the rate of recurrence of peritonitis is characteristically high. The cause of such a high rate of recurrence of peritonitis is probably a reflection of the predilection of Alcaligenes species to cause infection in the 'sicker' patients, and the almost universal resistance of this species to most antimicrobial agents. We, therefore, recommend that catheter removal be undertaken as early as the identification of the organism is made. Whether patients should be allowed to return to CAPD after recovery is a more difficult question. We suggest that a reevaluation of the patient's overall status be undertaken, including personal hygiene, exchange technique, presence of diabetes mellitus, malnutrition, and/or other factors that may render the patient more prone to infection with opportunistic pathogens. PMID- 9730576 TI - Extreme diuretic dependence in idiopathic edema: mechanisms, prevention and therapy. AB - Idiopathic edema patients abusing diuretics are occasionally becoming dependent to such a degree on increasing doses of diuretics that their withdrawal results in severe cardiorespiratory failure, occasionally even pulmonary edema. Two such patients are described and 1 is investigated in depth as to the mechanism of the diuretic abuse-induced excessive tubular avidity for sodium. An extreme diuretic induced secondary hyperaldosteronism and atrial natriuretic factor suppression, although tapering off when diuretics are stopped, results in a continuous tubular sodium hyper-reabsorption. Since the most affected patient was deprived of the benefits of converting enzyme inhibitors because of their side effects, the only way to partially overcome this condition was a generous combination of several diuretics acting at several segments of the nephron. This contrasted with a similar patient who was relatively well controlled by a converting enzyme inhibitor combined with lower dose diuretics. Diuretic abuse-induced secondary hyperaldosteronism and diuretic resistance are apparently best prevented by converting enzyme inhibitors. When nonpharmacological preventive measures fail, converting enzyme inhibitors are preferable to diuretics as the first-choice treatment of idiopathic edema patients. PMID- 9730577 TI - Brown tumor in association with secondary hyperparathyroidism. A case report and review of the literature. AB - Since 1993, our 34-year-old female patient was on replacement treatment with hemodialysis for terminal renal failure due to hypertensive nephropathy. She developed severe secondary hyperparathyroidism. In 1996, she presented with a brown tumor, localized in the region of the left maxillary sinus and at its front. The patient did not take phosphate binders and treatment with vitamin D was not possible due to the increased Ca x P product. She had no difficulties, so she did not desire a surgical parathyroidectomy. As she had trouble breathing through the nose, and because of the facial deformity due to the brown tumor, she decided to have the tumor removed surgically. PMID- 9730578 TI - Prolonged asymptomatic dense deposit disease in Chinese. Report of 2 cases in Taiwan. AB - Dense deposit disease (DDD) is a less common form of membranoproliferative glomerulonephritis (MPGN). The disease occurs predominantly in children and young adults and the prognosis is variable. DDD varies considerably in incidence among different populations and has not been reported in Chinese. Herein we reported 2 cases of DDD in young Chinese girls in Taiwan. Although 1 case (case 2) had mild hypertension, both patients had asymptomatic proteinuria and ran a benign course of 8 and 14 years, respectively. The histological features of case 1 resembled membranous glomerulonephritis (MGN) on hematoxylin-eosin stain, but revealed DDD on periodic acid Schiff and chromotrope-2R silver methenamine stains. Whereas case 2 showed focal MPGN on light microscopy, she had a fine granular immunofluorescence pattern resembling MGN. Characteristic intramembranous dense deposits were demonstrated by electron microscopy in the basement membranes of the glomeruli, Bowman's capsules and the renal tubules. Both patients were followed closely, and had stable normal renal function 1 year after renal biopsy. PMID- 9730579 TI - Abnormal adhesion of T cells to extracellular matrix proteins in hemodialysis patients. PMID- 9730581 TI - Major biological effects of neurotrophic factors on retinal ganglion cells in mammals. AB - The mammalian visual system, particularly retinal ganglion cells, has been used for studying the functions of neurotrophic factors on neurons for many years. The major biological effects of neurotrophic factors on retinal ganglion cells observed so far are the promotion of viability and axonal regeneration. However, there are still some controversies regarding the effects of neurotrophic factors on retinal ganglion cells in the literature. This review is aimed to summarize the available information on the biological actions of these neurotrophic factors on survival and axonal regeneration of retinal ganglion cells and the expressions of neurotrophic factor receptors in the retina. Generally, brain-derived neurotrophic factor, neurotrophin-4/5, fibroblast growth factor and glial cell line-derived neurotrophic factor increase the survival of retinal ganglion cells while the effect of ciliary neurotrophic factor on the viability of adult retinal ganglion cells is controversial. The ciliary neurotrophic factor is the only effective factor in promoting long distance axonal regeneration of retinal ganglion cells whereas brain-derived neurotrophic factor and neurotrophin-4/5 only enhance neurite sprouting within the retina. PMID- 9730580 TI - Prospects of the clinical utilization of melatonin. AB - This review summarizes the present knowledge on melatonin in several areas on physiology and discusses various prospects of its clinical utilization. Ever increasing evidence indicates that melatonin has an immuno-hematopoietic role. In animal studies, melatonin provided protection against gram-negative septic shock, prevented stress-induced immunodepression, and restored immune function after a hemorrhagic shock. In human studies, melatonin amplified the antitumoral activity of interleukin-2. Melatonin has been proven as a powerful cytostatic drug in vitro as well as in vivo. In the human clinical field, melatonin appears to be a promising agent either as a diagnostic or prognostic marker of neoplastic diseases or as a compound used either alone or in combination with the standard cancer treatment. Utilization of melatonin for treatment of rhythm disorders, such as those manifested in jet lag, shift work or blindness, is one of the oldest and the most successful clinical application of this chemical. Low doses of melatonin applied in controlled-release preparation were very effective in improving the sleep latency, increasing the sleep efficiency and rising sleep quality scores in elderly, melatonin-deficient insomniacs. In the cardiovascular system, melatonin seems to regulate the tone of cerebral arteries; melatonin receptors in vascular beds appear to participate in the regulation of body temperature. Heat loss may be the principal mechanism in the initiation of sleepiness caused by melatonin. The role of melatonin in the development of migraine headaches is at present uncertain but more research could result in new ways of treatment. Melatonin is the major messenger of light-dependent periodicity, implicated in the seasonal reproduction of animals and pubertal development in humans. Multiple receptor sites detected in brain and gonadal tissues of birds and mammals of both sexes indicate that melatonin exerts a direct effect on the vertebrate reproductive organs. In a clinical study, melatonin has been used successfully as an effective female contraceptive with little side effects. Melatonin is one of the most powerful scavengers of free radicals. Because it easily penetrates the blood-brain barrier, this antioxidant may, in the future, be used for the treatment of Alzheimer's and Parkinson's diseases, stroke, nitric oxide, neurotoxicity and hyperbaric oxygen exposure. In the digestive tract, melatonin reduced the incidence and severity of gastric ulcers and prevented severe symptoms of colitis, such as mucosal lesions and diarrhea. PMID- 9730582 TI - Adaptation-dependent differences in electroretinographic latency patterns in uniform and variegated horseshoe crabs. AB - The carapaces of horseshoe crabs (Limulus polyphemus) differ. Some individuals have uniform carapaces and clear eyes while others have variegated carapaces and dark eyes. These differences have been reported to be correlated with latency differences in the electroretinogram (ERG) of the lateral eye. Such a result might have had a neural basis in the mechanisms underlying visual transduction but it could also have reflected a visual screening pigment difference. A direct experiment was therefore designed to choose between these two hypotheses by varying the relative state of adaptation. The results were as follows. In uniform animals, dark adaptation had the kind of effect seen in single photoreceptor cells - latencies were longer in dark-adapted eyes and latencies were also longer for dim flashes. However, variegated animals showed a significant adaptation interaction: in light adaptation, dimmer flashes produced the usual effect, namely a longer ERG latency, while in dark adaptation, latencies were close to equilatent, being within experimental error of each other for both flash energies. These data make it unlikely that the photoreceptor transduction mechanism is the locus of the visual differences between the two types of animals. Instead, they are consistent with an interaction of screening pigment effects with photoreceptor transduction effects. PMID- 9730583 TI - Effects of amphetamine and phenylethylamine on catecholamine release in the glomerular layer of the rat olfactory bulb. AB - In the present work, we have shown electrochemically that in the rat olfactory bulb (OB), extracellular dopamine (DA) was highest in the glomerular layer (GL), whereas extracellular noradrenaline (NA) appeared to be more uniformly distributed across layers. The GL catecholamine (CA) responses to amphetamine (AMPH) and phenylethylamine (PEA) were also characterized electrochemically using an in vivo model. Results of this investigation show that at a lower dose (1 mg/kg), PEA had no effect on CA release. In contrast, at a higher dose (10 mg/kg), it produced similar increases in either extracellular DA (17.5 +/- 7%) or extracellular NA (14 +/- 3%), and DA exhibited dose-independent increases to AMPH (93 +/- 8%: 1 mg/kg vs. 97 +/- 6%: 10 mg/kg) whereas NA exhibited dose-dependent increases to AMPH (24.5 +/- 6%: 1 mg/kg vs. 39 +/- 7%: 10 mg/kg). These data indicate that (i) PEA may increase CA release but less efficiently than AMPH. (ii) AMPH is more efficient on the DAergic than on the NAergic system since AMPH induced DA release exceeded 2-4 times the AMPH-induced NA release. PMID- 9730584 TI - An updated phylogenetic analysis of vertebrate melatonin receptor sequences: reflection on the melatonin receptor nomenclature by the Nomenclature Subcommittee of the International Union of Pharmacology. AB - In the past few years, significant progress on melatonin receptor research has led to the discovery of a family of genetically related but pharmacologically distinctive G-protein-coupled receptors in the vertebrates. With increasing number of receptor clones being identified, there is a need for a system of classification and nomenclature for these receptor subtypes. Recently, an updated nomenclature system, which has renamed the existing mammalian melatonin receptor clones, has been proposed by the relevant subcommittee of the International Union of Pharmacology (NC-IUPHAR). However, the majority of receptor clones which have been identified in non-mammalian vertebrates are not clearly defined by this system. By performing phylogenetic analysis of both mammalian and non-mammalian melatonin receptor clones, we would like to propose a classification-nomenclature system for vertebrate melatonin receptors. Hopefully, our system, which incorporates genetic data as well as the pharmacological criteria that have been adopted by the NC-IUPHAR nomenclature system, will provide the framework for future development of a unified scheme of classification and nomenclature for melatonin receptors. PMID- 9730585 TI - Hot topics in and new strategies for surfactant research. PMID- 9730586 TI - Synthetic surfactant protein analogues. AB - Surfactant preparations for the treatment of respiratory distress syndrome (RDS) that contain phospholipids and small amounts of the two hydrophobic proteins, SP B and SP-C, are presently obtained from animal lungs. Since structural information about SP-B and SP-C is available, it appears possible to design analogues that can replace the native proteins in synthetic surfactants. SP-C contains a single helix, but analogues with the poly-Val sequence of the native molecule do not fold into a native-like alpha-helical conformation. However, replacement of all Val with Leu yields efficient folding into a helical structure and Leu-based SP-C analogues effectively accelerate spreading of surfactant lipids and exhibit some physiological activity in animal models of RDS. The inferior in vivo activity of synthetic surfactants containing SP-C only compared to that of surfactant preparations derived from natural sources may be caused by a lack of covalently linked palmitoyl groups in the analogues and/or absence of SP-B. SP-B is significantly larger than SP-C and has a tertiary fold of several amphipathic helices in a dimeric structure. A single simplified amphipathic helical peptide containing only Leu and Lys does not mimic the surface properties of SP-B in vitro. These circumstances make the design of SP-B analogues from solely structural considerations less likely to be successful than in the case of SP-C. PMID- 9730587 TI - Surfactant therapy in acute respiratory distress syndrome. PMID- 9730588 TI - Chronic lung disease: the role of oxidative stress. PMID- 9730589 TI - Superoxide dismutase: a role in the prevention of chronic lung disease. PMID- 9730591 TI - Abstracts PMID- 9730590 TI - Surfactant treatment of the very preterm infant. PMID- 9730592 TI - Report of the Fourth International Workshop on Human Chromosome 5 mapping 1996. PMID- 9730593 TI - Assignment of the hepatocyte nuclear factor 3 genes to rat chromosome bands 6q23- >q24 (alpha: Hnf3a), 3q41 (beta: Hnf3b) and 1q21-->q22 (gamma: Hnf3g) by in situ hybridization. PMID- 9730595 TI - Assignment of a human putative RNA helicase gene, DDX3, to human X chromosome bands p11.3-->p11.23. PMID- 9730594 TI - Assignment of AR1, transcription factor 20 (TCF20), to human chromosome 22q13.3 with somatic cell hybrids and in situ hybridization. PMID- 9730596 TI - Assignment1 of H7365 (C9orf2) to human chromosome band 9q31 by somatic cell hybrid analysis and fluorescence in situ hybridization. PMID- 9730597 TI - Assignment of Erbb2 to rat chromosome band 10q32.1 by in situ hybridization. PMID- 9730598 TI - FHIT gene transcript alterations occur frequently in myeloproliferative and myelodysplastic diseases. AB - Twenty-five primary biopsy samples, obtained from patients diagnosed with chronic/acute myeloproliferative disorders, myelodysplastic disorders, in addition to seven cell lines established from patients with leukemias arrested at different stages of myeloid differentiation, were examined for alterations in an alternatively spliced form of the FHIT gene. Transcript alterations of this gene were detected in 80% of the primary biopsies and in two of the cell lines. Reverse transcription PCR (RT-PCR) detected deletions of one or more specific exons in the translated or untranslated portion of the FHIT gene. Point mutations in exons 3, 4, and 5 of the FHIT gene were also detected in 4 patients by single stranded conformational PCR analysis. Transcript alterations were detected in all primary hematopoietic samples which contained chromosome abnormalities, as well as, in hematopoietic disorders which did not show alterations of the 3p14 region. These findings suggest FHIT gene involvement in the transformation of hematopoietic stem cells to leukemia. PMID- 9730599 TI - Assignment1 of the Smad7 gene (MADH7) to human chromosome 18q21.1 by fluorescence in situ hybridization. PMID- 9730600 TI - Cloning and mapping of SMARCA5 encoding hSNF2H, a novel human homologue of Drosophila ISWI. AB - We have isolated a novel cDNA encoding a peptide with 86% sequence homology to hSNF2L protein, a previously isolated human homologue of Drosophila ISWI. This gene, designated SMARCA5, contained an open reading frame of 3,156 nucleotides encoding a 1,052 amino-acid peptide (hSNF2H). As this product also revealed a significant (73%) identity in amino acid sequence to ISWI, a key component of chromatin-remodeling factors in Drosophila, hSNF2H may be another human homologue of this protein and, as such, could be involved in chromatin remodeling in humans. An ATPase domain characteristic of the SWI2/SNF2 family of proteins was highly conserved in ISWI, hSNF2L, and hSNF2H. Northern-blot analysis demonstrated ubiquitous expression of 5.1-kb and 4.1-kb transcripts of the hSNF2H gene. This gene was mapped by FISH to chromosome bands 4q31.1-->q31.2. PMID- 9730601 TI - Cloning and genomic mapping of the mouse matrin 3 gene and its pseudogenes. AB - Matrin 3 forms a family of nuclear proteins together with NP220s. Using cDNA sequences encoding rat matrin 3 as a probe, we isolated genomic clones of mouse matrin 3 gene and its two pseudogenes. The genuine mouse matrin 3 gene has an exon covering the N-terminal one third of matrin 3 with a sequence 99% identical to rat matrin 3 cDNA. The gene was localized to Chromosome 18C by in situ hybridization and mapped at 3.6 cM distal to D18Mit117 and 2.2 cM proximal to D18Mit14 on mouse Chromosome 18 by interspecific backcross analysis. One pseudogene has an exon-like sequence covering the N-terminal half of matrin 3 that is interrupted by an unknown sequence. Reverse transcription polymerase chain reaction (RT-PCR) of RNA from mouse liver and brain using unique sequences of the genuine gene and the pseudogene confirmed the expression of only the former. The other pseudogene has a sequence only 80% identical to rat cDNA and is partially deleted and reversed. These pseudogenes were localized to Chromosome 4E2 and 8D2, respectively. PMID- 9730602 TI - Identification and in situ hybridization mapping of a mouse Tpd52l1 (D53) orthologue to chromosome 10A4-B2. AB - We report the identification of a mouse cDNA Tpd52l1 (tumor protein D52-like 1), which represents the first demonstrated orthologue of the human TPD52L1 (alias D53) gene, a member of the breast carcinoma-associated TPD52 (alias D52) gene family. In situ hybridization mapping located the Tpd52l1 gene to chromosome 10A4 10B2. Since the TPD52L1 gene is found at human chromosome 6q22-->q23, the mouse and human TPD52L1 loci are syntenically conserved. PMID- 9730603 TI - Comparative FISH-mapping of the prion protein gene (PRNP) on cattle, river buffalo, sheep and goat chromosomes. AB - Comparative FISH-mapping of the prion protein gene (PRNP) was performed on cattle (BTA), river buffalo (BBU), sheep (OAR) and goat (CHI) chromosomes using a PCR product as a probe and R-banding. PRNP was mapped to BTA13q17, BBU14q15, OAR13q15 and CHI13q15 according to standard nomenclatures. These chromosomes and bands were homoeologous among the four species, confirming the high degree of gene and chromosome banding conservation among bovids. Furthermore, the assignment of PRNP to river buffalo and goat chromosomes allowed us to indirectly assign the bovine syntenic group U11 to specific chromosomes, since it is the first in situ localization on BBU14 and CHI13. PMID- 9730604 TI - Physical and linkage mapping of the gene for the alpha3 chain of type IX collagen, COL9A3, to human chromosome 20q13.3. AB - Type IX collagen is a minor cartilage component which associates with mixed fibrils of types II/XI collagen. We have determined the precise physical and genetic locations for the gene encoding the alpha3 chain of type IX collagen, COL9A3. Utilizing fluorescence in situ hybridization, radiation hybrid mapping, and multipoint linkage analysis, we have mapped COL9A3 to human chromosome 20q13.3, 13 cM telomeric to D20S173. PMID- 9730605 TI - Comparative FISH mapping of mouse and rat homologues of twenty-five human X linked genes. AB - We constructed a comparative cytogenetic map of 25 functional genes in mouse and rat X chromosomes by direct R-banding fluorescence in situ hybridization. Nineteen and 22 out of the 25 genes, which have been mapped on the human X chromosome, were newly localized to mouse and rat X chromosomes, respectively. Twenty-two additional genes were integrated in the rat-mouse-human comparative map of the X chromosome in this study. Comparison of the gene order indicated the presence of four chromosome segments with conserved linkage homology between mouse and rat X chromosomes, suggesting that a minimum of four chromosomal inversion events occurred between mouse and rat X chromosomes during the evolution of the two species. Four chromosome segments with conserved linkage homology were found between human and rat X chromosomes. PMID- 9730606 TI - Assignment of the mouse and cow CXC chemokine genes. AB - Gene specific PCR primers were constructed for five mouse and three bovine CXC chemokine genes. The mouse genes were assigned using SSCP analyses of the Jackson BSS backcross panel to two groups on chromosome 5. One group containing Gro1 and Mip2 cosegregated with reference markers Alb1 and Btc, and was positioned 2.2 cM proximal to a group comprising Ifi10, Mig, and Scyb5. The bovine genes IL8, GRO1, and GRO3, mapped using bovine x hamster somatic cell hybrids, were all found to be located on chromosome 6. The locations of these genes in these two animal species are consistent with the positions in humans (4q13-->q21), and previous syntenic relationships among these three mammals. PMID- 9730608 TI - Cat eye syndrome chromosome breakpoint clustering: identification of two intervals also associated with 22q11 deletion syndrome breakpoints. AB - The supernumerary cat eye syndrome (CES) chromosome is dicentric, containing two copies of 22pter-->q11.2. We have found that the duplication breakpoints are clustered in two intervals. The more proximal, most common interval is the 450 650 kb region between D22S427 and D22S36, which corresponds to the proximal deletion breakpoint interval found in the 22q11 deletion syndrome (DiGeorge/velocardiofacial syndrome). The more distal duplication breakpoint interval falls between CRKL and D22S112, which overlaps with the common distal deletion interval of the 22q11 deletion syndrome. We have therefore classified CES chromosomes into two types based on the location of the two breakpoints required to generate them. The smaller type I CES chromosomes are symmetrical, with both breakpoints located within the proximal interval. The larger type II CES chromosomes are either asymmetrical, with one breakpoint located in each of the two intervals, or symmetrical, with both breakpoints located in the distal interval. The co-localization of the breakpoints of these different syndromes, plus the presence of low-copy repeats adjacent to each interval, suggests the existence of several specific regions of chromosomal instability in 22q11.2 which are involved in the production of both deletions and duplications. Since the phenotype associated with the larger duplication does not appear to be more severe than that of the smaller duplication, determination of the type of CES chromosome does not currently have prognostic value. PMID- 9730607 TI - Cytogenetic localization of cancer-related genes in the rat and comparative mapping studies in human and mouse. AB - The rat is an important model organism in biomedical research, and several well characterized rat cancer model systems exist. To facilitate detailed analysis of these models, it is useful to know the regional location of known cancer-related genes. In this report, 14 cancer-related genes have been sublocalized by fluorescence in situ hybridization. The mapped genes include three oncogenes (Fyn, Mas1, and Vav1), a tyrosine kinase gene (Syk), a tumor-associated antigen gene (Cd24), a growth factor receptor gene (Igf2r), the gene for an activator of c-fos/c-jun transcription factors (Apex), a transcription factor gene (Egr3), and several genes involved in steroid hormone metabolism and signaling (Esr2, Pgr, Cbg, Cyp17, and Cyp19). The remaining gene (Map1a) is involved in microtubule assembly. PMID- 9730609 TI - Physical mapping of the t(12;22) translocation breakpoints in a family with a complex type of 3/3'/4 synpolydactyly. AB - We previously reported clinical and radiological findings in a Belgian family with a complex type of synpolydactyly associated with metacarpal and metatarsal synostoses, cosegregating with a balanced t(12;22). Recently, expansions of a polyalanine stretch within the first exon of the HOXD13 gene, which resides on chromosome 2q31, have been shown to cause synpolydactyly (SPD). Using exon amplification followed by direct sequencing, we were able to exclude the direct involvement of the HOXD13 gene in this family. As a first step toward the positional cloning of a candidate disease gene on chromosome 12 and/or 22 responsible for the type of complex synpolydactyly observed in this family, we report here the construction of a somatic cell hybrid retaining only the der(22) of the t(12;22)(p11.3;q13.3). STS content mapping and FISH experiments allowed us to position the chromosomal breakpoints between markers D12S1596 and D12S1034 on chromosome 12 and markers N73F4 and D22S158 on chromosome 22. PMID- 9730610 TI - Assignment of the human erythrocyte acylphosphatase gene (ACYP1) to chromosome band 14q24.3. PMID- 9730612 TI - Expressed copies of the MN7 (D15F37) gene family map close to the common deletion breakpoints in the Prader-Willi/Angelman syndromes. AB - Approximately 70% of patients with Prader-Willi syndrome or Angelman syndrome have a similar sized de novo deletion of 3-4 Mb in the proximal region of 15q. The distal breakpoints appear to cluster between the P gene (OCA2) and D15S24, whereas two deletion breakpoint clusters have been identified on the proximal side (one centromeric to D15S541 and one between D15S541 and D15S9). Based on the identification of a gene family in 15q11-->q13 (MN7, D15F37), we have previously proposed that the presence of multiple copies of this sequence may be related to the instability of this region. Using fluorescence in situ hybridization and YAC mapping, we have found that at least one D15F37 locus is centromeric to D15S9 and at least two are between OCA2 and D15S24. As determined by cDNA cloning and sequence analysis, each of the individual loci is expressed. The close proximity of the D15F37 loci and the deletion breakpoints suggests that the common deletions arise by unequal crossover events at or near these loci. PMID- 9730611 TI - Comparative mapping of two adjacent regions of MMU19 with their human counterpart on HSA11q13. AB - High resolution physical maps of two adjacent regions of MMU19 were constructed in order to establish a comparative map between the pericentromeric region of MMU19 and its human counterpart on HSA11q13. These two physical maps span 2.5 and 0.5 megabases on MMU19. Long range restriction analysis and YAC contigs have been built, five genes were located on MMU19 and eight new STSs were generated. The 0.5-Mb map which has been positioned close to the centromere of MMU19, based on dual-color FISH experiments and genetic data, includes eight genes (Type I markers), three microsatellites (Type II markers) and five new STSs. The 2.5-Mb map is located more telomeric and contains seven genes, four microsatellites and four new STSs. Gene order and physical distances appear to be similar in human and in mouse in this 2.5-Mb region. Strikingly, the 0.5-Mb region has a similar size in human but gene order is shuffled. The overall comparative map shows that these two regions are inverted on MMU19 when compared with HSA11q13. PMID- 9730613 TI - Chromosome identification in human oocytes and polar bodies by spectral karyotyping. AB - Sixty unfertilized human oocytes and two fresh polar bodies were karyotyped by spectral karyotyping (SKY). The oocytes were provided by 29 women ranging from 30 to 42 yr of age. The mean hybridization efficiency for oocytes was 95.2% (60/63). Nondisjunction of bivalent chromosomes (13.3%) and predivision of sister chromatids at meiosis I (3.3%) were unequivocally determined by analysis first with SKY and then fluorescence in situ hybridization. Four oocytes (6.7%) were hyperhaploid, six (10.0%) were hypohaploid, one (1.7%) showed balanced predivision, and another (1.7%) was diploid. No specific structural rearrangements were detected. This study demonstrates that the SKY technique can be used successfully as an alternative method of karyotyping second meiotic metaphase chromosomes from human oocytes and polar bodies in appropriate spreads. PMID- 9730614 TI - High-resolution mapping of the X-linked lymphoproliferative syndrome region by FISH on combed DNA. AB - X-linked lymphoproliferative syndrome is an inherited immunodeficiency for which the responsible gene is currently unknown. Several megabase-sized deleted regions mapping to Xq25 have been identified in XLP patients, and more recently a 130-kb deletion has been reported (Lamartine et al., 1996; Lanyi et al., 1996). To establish a physical map of this deleted region and to identify the XLP gene, two cosmid contigs were established (Lamartine et al., 1996). However, the physical map of this region is still uncompleted and controversial and three points remain unsolved: (1) the centromeric-telomeric orientation of the whole region, (2) the relative orientation of the two contigs, and (3) the size of the gap between the two contigs. To provide a definitive answer to these questions, high-resolution mapping by fluorescence in situ hybridization on combed DNA and molecular approaches were combined to establish the physical map of the XLP region over 600 kb. Our results identified a gap of 150 kb between the two contigs, established the relative orientation of one contig to the other, and determine the centromeric-telomeric orientation of the whole region. Our results show that the order of the marker over this region is: cen.1D10T7-DF83-DXS982.tel. PMID- 9730615 TI - Murine and human TSPYL genes: novel members of the TSPY-SET-NAP1L1 family. AB - Using expressed sequence tag (EST) sequence information, novel genes related to the Y chromosome gene family TSPY were isolated and characterized. Because of a significant homology to TSPY, the novel genes were designated TSPY-Like (TSPYL) and were assigned as new members of the TSPY-SET-NAP1L1 family. A human TSPYL gene was localized on chromosome 6 and a murine Tspyl gene was localized on chromosome 10. Expression of TSPYL was observed in all tissues investigated, as well as early onset during development. Both the human and murine Tspyl homologs lack introns over the entire region so far investigated and are thought to have arisen by an ancient retroposition event. Retroposition of Tspyl genes is supported by the isolation of a murine Tspyl pseudogene on chromosome 12 which also lacks intronic sequences, and by its observed proximity to an R element, a family of dispersed repetitive DNA. PMID- 9730616 TI - The human gene encoding SCYB9B, a putative novel CXC chemokine, maps to human chromosome 4q21 like the closely related genes for MIG (SCYB9) and INP10 (SCYB10). PMID- 9730617 TI - Assignment1 of Sp genes to rat chromosome bands 7q36 (Sp1), 10q31-->q32.1 (Sp2), 3q24-->q31 (Sp3) and 6q33 (Sp4) and of the SP2 gene to human chromosome bands 17q21.3-->q22 by in situ hybridization. PMID- 9730618 TI - The INSL4 gene maps close to WI-5527 at 9p24.1-->p23.3 clustered with two relaxin genes and outside the critical region for the monosomy 9p syndrome. AB - The insulin like growth factor 4 (INSL4) gene belongs to the insulin gene superfamily and has been mapped by fluorescent in situ hybridization to 9p24. Expression of INSL4, of unknown function, has been recently detected in the perichondrium and ligaments of the human embryo. Here we have mapped INSL4 within the framework of a partial YAC contig covering the distal part of 9p to find out whether this gene lies within the critical region defined for the monosomy 9p syndrome. The gene was also located using a human-rodent radiation hybrid panel. INSL4 was found to be distal to the del(9p) critical region and excluded as a candidate for the syndrome. In addition, the positions of two relaxin genes (RLN1, RLN2) that belong to the same superfamily as INSL4 and which have also been mapped to chromosome 9, were refined. We have shown that the three genes are clustered in the same region. PMID- 9730619 TI - Human serine/threonine protein kinase EMK1: genomic structure and cDNA cloning of isoforms produced by alternative splicing. AB - The EMK (ELKL Motif Kinase) is a small family of ser/thr protein kinases involved in the control of cell polarity, microtubule stability and cancer. We have isolated several cDNA clones that encoded two isoforms of the human ser/thr protein kinase EMK1 (GDB symbol EMK1, alias ELKL motif kinase 1, and MARK2). These isoforms were characterized by the presence of a 162-bp alternative exon that gave rise to two forms, one containing the exon and the other one lacking it. Both forms were found to be coexpressed in a number of selected cell lines and tissue samples when analyzed by RT-PCR. By Northern blot, human EMK1 was shown to be encoded by a single mRNA ubiquitously expressed. We have reconstructed most of the genomic structure of EMK1 from the sequence of two human chromosome 11 cosmids deposited in databases, and showed that the gene is composed of a minimum of 16 small exons. PMID- 9730620 TI - Assignment of the ZYX gene for the LIM protein zyxin to human chromosome bands 7q34-->q35 by in situ hybridization. PMID- 9730621 TI - Assignment of the BMPR1A and BMPR1B genes to human chromosome 10q22.3 and 4q23- >q24 byin situ hybridization and radiation hybrid map ping. PMID- 9730622 TI - Gene structure and chromosome location of mouse Cd39 coding for an ecto-apyrase. PMID- 9730623 TI - Assignment of the cyclin-dependent kinase inhibitor genes Cdkn2a and Cdkn2b to rat chromosome bands 5q32-->q34 and 5q31-->q33, respectively by fluorescence in situ hybridization, using small PCR-generated probes. PMID- 9730624 TI - Assignment of human fatty-acid-coenzyme A ligase, very long-chain 1 gene (FACVL1) to human chromosome band 15q21.2 by fluorescence in situ hybridization. PMID- 9730625 TI - Assignment of Sox4 to mouse chromosome 13 bands A3-A5 by fluorescence in situ hybridization; refinement of the human SOX4 location to 6p22.3 and of SOX20 to chromosome 17p12.3. PMID- 9730653 TI - Imaging approaches to the correlation of structure and function in the kidney. AB - AIMS: This review sets out to cover the areas of radiology and imaging concerned with the structure and function of the kidney which may not be familiar to those outside the field. METHODS: A brief historical introduction to renal imaging is followed in more detail by descriptions of the use of computed tomography in 3D reconstruction, angiography, and functional studies. Comparison with magnetic resonance imaging is made and the use of radiological techniques in intervention. CONCLUSION: Radiology can provide far more information about the kidneys than mere anatomy. Research promises the possibility of reducing the number of tests required to provide structural and functional information. PMID- 9730654 TI - Scintigraphic imaging of renal function. AB - Scintigraphic imaging of renal physiology can be achieved by gamma camera planar imaging, single photon emission computerised tomography (SPECT) or positron emission tomography (PET). Physiological information about the kidney usually requires dynamic imaging, although for some applications imaging may be static. Individual kidney renal blood flow can be measured from dynamic gamma camera imaging, although with PET it is possible to measure regional renal perfusion. Measurement of individual kidney glomerular filtration rate (GFR) is performed routinely with Tc-99m-diethylene triamine pentaacetic acid (DTPA), but information about GFR on a regional basis, and measurement of regional filtration fraction, require PET. Manoeuvres which interfere with renal physiology, such as captopril renography, may also yield useful diagnostic information. PMID- 9730655 TI - Conventional and confocal epi-reflection and fluorescence microscopy of the rat kidney in vivo. AB - To visualize superficial and accessible renal tubule cells functioning in situ and to relate what we can 'see' to what we know of their function from more invasive in vivo or less direct in vitro studies means applying and adapting recent advances in epifluorescence and confocal microscopy to improve image resolution and to combine this with the use of fluorescent labels to monitor the handling of specific molecules by the proximal and distal renal tubule cells in vivo. Doing this in living tissue is novel, especially in the kidney. Application of confocal microscopy to the imaging of living tissue, as opposed to isolated cells, has not been widely reported. The kidney surface has been imaged before using the confocal microscope and in preliminary studies we have extended this by using a different confocal system with and without fluorescence. While the studies published up to now have been morphological, comparing standard renal (structural) histology of surface glomeruli and renal tubules with the corresponding in vivo confocal images, more dynamic, real-time studies have been limited. Individual red blood cells can be seen flowing around the peritubule capillary network and nucleated white blood cells can also be distinguished. Tubule cells, endothelial cells, the proximal tubule cell brush border and cell mitochondria can be visualized. Filtration and secretion can be observed, and the early and late parts of the proximal tubule distinguished, and the distal tubule recognized. Localization of fluorescently labeled insulin to the luminal brush border and progressive uptake of label and distribution within proximal tubule cells toward the basolateral (blood side) membrane can be demonstrated. The possibility of monitoring hemodynamic changes and tracking the filtration, uptake, secretion and absorption of fluorescently tagged molecules, as well as intracellular fluorescence, e.g. calcium or pH, is an exciting prospect and is ripe for detailed exploration. PMID- 9730656 TI - Nuclear magnetic resonance spectroscopy: a non-invasive probe of kidney metabolism and function. AB - Major advances in nuclear magnetic resonance (NMR) spectroscopic methods and technology have led to the increased use of this technique to study kidney metabolism and function. These studies include: (1) the identification of organic osmolytes in the renal medulla and their role as potential markers of medullary development and damage; (2) changes in renal epithelial cell organic solute transport, such as autosomal dominant polycystic kidney disease, and (3) the biochemical heterogeneity of the nephron and identification of markers of site specific renal damage in experimental animals and man. The present review summarises these data with the aim of demonstrating how NMR can be used as an indirect, and non-invasive probe of homeostatic mechanisms in vivo and in vitro. PMID- 9730657 TI - Applications of capillary electrophoresis in nephrology. AB - Capillary electrophoresis has recently emerged as a powerful technique for separating components in biological samples. A family of separation methods capable of handling a diverse range of samples has been developed, the sample volumes required are very small and a wide range of specialised detectors can be employed. This review examines some methods with particular application to the analysis of urine and tubular fluid samples and references relevant applications. PMID- 9730658 TI - Ultramicroanalysis of peptide profiles in biological samples using MALDI mass spectrometry. AB - In the past few years, matrix-assisted laser desorption/ionization (MALDI) mass spectrometry has emerged as a powerful tool for the direct analysis of peptide profiles contained in single neuroendocrine cells, tissue biopsies, as well as in releasates (after a simple sample clean-up). These studies were performed in both invertebrate (the pond snail) and vertebrate (xenopus and rat) species. The present article provides an overview of these data with a special emphasis on the sample handling required for each application. PMID- 9730659 TI - Gene transfer into the adult kidney for unravelling disease processes. AB - Insights into the pathogenesis of human disease must be based on an understanding of the molecular mechanisms that regulate the structure and function of individual organs. For this purpose, gene transfer technologies provide powerful and attractive tools. In principle, two different approaches are feasible to identify pathophysiological roles of certain genes: 'gain-of-function', and 'loss of-function'. The former examines consequences of overexpression of an exogenous gene, and the latter investigates the outcomes of inhibition of a particular molecule via antisense, decoy, ribozyme and dominant-negative strategies. Gene transfer to specific renal structures allows evaluation of in vivo effects of certain molecules on the structure and function of each nephron segment. It would also be useful for therapeutic intervention in renal diseases through introduction of therapeutically relevant genes into affected sites. This article summarizes current experience with renal gene transfer and addresses its potential impacts on the understanding of renal function in vivo. PMID- 9730660 TI - Transgenic models in renal tubular physiology. AB - Animal transgenesis has proven to be useful for physiological as well as physiopathological studies. Besides the classical approach based on the random integration of a DNA construct in the mouse genome, gene targeting can be achieved using totipotent embryonic stem (ES) cells for targeted transgenesis. Transgenic mice are then derived from the transgenic ES cells. This allows the introduction of null mutations in the genome (so-called knock-out) or the control of the transgene expression by the endogenous regulatory sequences of the gene of interest (so-called knock-in). Development of these transgenic animals leads to a better understanding of the cellular function of many genes or to the generation of animal models for human diseases. The purpose of this short review is to describe animal models in renal tubular physiopathology. Recent progresses will allow the generation of animal models with conditional expression of the transgene of interest or with a conditional gene mutation. This permits spatial and temporal control of the expression of the transgene or of the mutation. This should allow the generation of models suitable for physiological analysis or closer to disease state. PMID- 9730661 TI - Tubule function in transgenic mice. AB - Many transporters involved in renal electrolyte transport have recently been identified and cloned. The availability of genetically manipulated mice, especially transgenic knockout and overexpression models, has made it possible to examine ion transport in kidney tubules in the absence of specific transporters whose function in defined tubule segments is well known. Such selective alterations in transport functions are also useful to investigate adaptive mechanisms by which the kidney compensates for specific transport lesions. Examples of mouse models displaying altered renal transport function include targeted disruption of genes encoding the Na-H exchanger isoforms NHE2 and NHE3, the thiazide-sensitive Na-Cl cotransporter TSC, CFTR, and the colonic isoform of the H,K-ATPase. Moreover, mice with null mutation in the Na-H exchanger isoform NHE1 have been also identified. In addition, a strain of mice with enhanced H,K ATPase expression due to a defective endocytosis signal has been developed. Other transporter knockout models will soon become available. In this review we focus on the physiological characterization of renal tubule transport in animals with well-defined genetic transport lesions. PMID- 9730662 TI - Combined in vivo and in vitro approaches to analysis of renal tubule function. AB - The study of renal tubule function obtained its first impulse with whole kidney approaches, such as the clearance technique, and is presently centered on cellular analysis involving intracellular ion activities and on molecular studies describing the structure of transporter molecules and the function of their subunits and constitutional building blocks as well as their distribution along the nephron and within individual cells. Between these extremes a number of successful techniques have dominated the field, including the micropuncture methods which were pioneered by Richards, Walker and others in the late 1920s and 1930s, and were revived and perfectioned in the following decades leading to complex in vivo and in vitro micromethods that for an important period of time have been the center of scientific progress in this area. In the present paper, some of the methods in this field are shortly reviewed, specifically from the point of view of acid-base physiology applied to the renal tubule. PMID- 9730663 TI - Insights from mathematical modeling of renal tubular function. AB - Mathematical models of proximal tubule have been developed which represent the important solute species within the constraints of known cytosolic concentrations, transport fluxes, and overall epithelial permeabilities. In general, model simulations have been used to assess the quantitative feasibility of what appear to be qualitatively plausible mechanisms, or alternatively, to identify incomplete rationalization of experimental observations. The examples considered include: (1) proximal water reabsorption, for which the lateral interspace is a locus for solute-solvent coupling; (2) ammonia secretion, for which the issue is prioritizing driving forces - transport on the Na+/H+ exchanger, on the Na,K-ATPase, or ammoniagenesis; (3) formate-stimulated NaCl reabsorption, for which simple addition of a luminal membrane chloride/formate exchanger fails to represent experimental observation, and (4) balancing luminal entry and peritubular exit, in which ATP-dependent peritubular K+ channels have been implicated, but appear unable to account for the bulk of proximal tubule cell volume homeostasis. PMID- 9730664 TI - Intrauterine growth restriction: definition and etiology. AB - Intrauterine growth restriction (IUGR) is a frequent cause of perinatal morbidity as well as of impaired growth during childhood. Therefore, a clearcut definition of IUGR to identify those babies at risk is essential: The label IUGR generally should be assigned only to those infants with birth weight and/or birth length below the 10th percentile for GA with a pathologic restriction of fetal growth. According to the recent literature, clinical classification of the retarded babies seems to be less significant. Among the etiologic factors responsible for IUGR, one-third of the variations in birth weight are determined by genetic variables, two-thirds by environmental factors. In spite of the fact that a long list of established, different etiologic factors is known, in at least 40% of children no underlying pathology can be identified. Among the preventable, environmental causes of IUGR, smoking of the mother during pregnancy is by far the most important one, which is responsible for more than one third of all IUGR newborns. PMID- 9730665 TI - Children born small-for-gestational age: postnatal growth and hormonal status. AB - It is generally recognized that children born small-for-gestational age (SGA) have a 5-7 times higher risk of short stature than children born at normal size. It has been suggested that the programming of the endocrine axes occurs during critical phases of fetal development and is affected by intrauterine growth retardation. This study was undertaken to characterize the postnatal growth pattern and the final height of children born SGA, as part of a population- based study (n = 3,650), from birth to final height, and to evaluate the hormonal status in another group of prepubertal children born SGA (n = 134) without postnatal catch-up growth. The majority (88%) of 'healthy' full-term singleton SGA infants achieved catch-up growth during the first 2 years of life, and most of the increase in height occurred by 2 months of age. The SGA children who remained short at 2 years of age had a higher risk of short stature later in life. The risk of having a short final height (<-2 SDS) was five times higher for children with a low birth weight and seven times higher for those with a low birth length in comparison with children with a normal birth size. Moreover, about 20% of all children of short stature were born SGA. As a group, children born SGA will have a final height, expressed in SDS, as they had during the prepubertal years. This is in contrast to children, who became short postnatally. During puberty, these short children will have a mean height gain of 0.6 SDS for girls and 0.7 SDS for boys. The mean estimated secretion rate for growth hormone (GH) was lower in the short children born SGA compared with the reference groups born at an appropriate size for gestational age, of either short (p < 0.05) or normal stature (p < 0.001). Moreover, in the youngest children born SGA (2-6 years of age) a different pattern of GH secretion was found, with a high basal GH level, low peak amplitude, and high peak frequency. The majority of the children born SGA had levels of GH-binding protein within the range previously reported for normal children. However, the levels of insulin-like growth factor I (IGF-I), IGF-binding protein-3 (IGFBP-3) and leptin were significantly reduced compared with the reference values (p < 0.001, p < 0.01 and p < 0.001, respectively). In conclusion, the low spontaneous GH secretion rate and a disturbed GH secretion pattern, together with low serum levels of IGF-I, IGFBP-3 and leptin, might contribute to the reduced postnatal growth in some of the subgroup of children born SGA who remained short during childhood. PMID- 9730666 TI - Diagnosis and management of intrauterine growth retardation. AB - Intrauterine growth retardation (IUGR) is associated with significant perinatal morbidity and mortality. This condition can be a sign of genetic disorders, fetal infection, uteroplacental insufficiency, or constitutionally small fetuses. Correct determination of gestational age is the first step in prenatal screening of growth-retarded fetuses. The discovery of a small-for-gestational age fetus necessitates fetal assessment for the evaluation of the etiology and prognosis, and for the determination of the optimal timing for delivery of these fetuses at risk of perinatal asphyxia. IUGR is more frequent in multiple-gestation pregnancies (23-34%) and will be discussed separately. There is no medical treatment for IUGR. Early aspirin treatment reduces the incidence of IUGR in a high-risk population but should not be used routinely in all pregnant women. PMID- 9730667 TI - Multiple hormone resistance in short children born with intrauterine growth retardation? AB - Intrauterine growth retardation (IUGR) is encountered in 2.5% (-2 SD) of newborns. Lack of postnatal catch-up growth is found in 8-20%. If GH secretion is increased early postnatally in IUGR, then some persistently short IUGR children may present with GH insufficiency. However, the mechanism of postnatal catch-up growth is heterogenous. The response to GH treatment with regard to plasma IGF-1, GH dose and growth velocity was analyzed in persistently short idiopathic IUGR children and compared to GH-deficient (GHD) and familial short stature (FSS) children of similar age and degree of short stature. IUGR children require both a greater basal and GH-induced plasma IGF-1 in order to achieve a growth velocity of similar magnitude to that of FSS and GHD children. These data suggest a different sensitivity to GH in IUGR compared to FSS or GHD children, sustaining the hypothesis that these idiopathic IUGR children may be partially IGF-1 resistant. The recent report of partial insulin resistance in IUGR subjects raises the possibility of an IGF-1 receptor- or post-receptor-mediated defect. PMID- 9730668 TI - Growth hormone in the treatment of short stature in young children with intrauterine growth retardation. AB - Growth acceleration and bone maturation were studied in children with severe short stature and a history of intrauterine growth retardation (IUGR) to determine the effect of growth hormone (GH)-treatment. Patients were enrolled in an open randomized, multicenter trial and allocated to either the treated group (group 1, n = 38) or the control group (group 2, n = 31). Both groups were treated daily with 0.2 IU/kg/body weight during 3 years. The children in group 2 started the study with a 1-year observation period followed by a 3-year treatment period. At baseline, the mean age was 4.5 years, bone age was 3.3 years, height standard deviation score (SDS) was -3.4, height velocity (HV) SDS was -1.6. Auxologic data were measured every 3 months and bone age was assessed at the start of the study (baseline) and then every 12 months thereafter. After 1 year of treatment, linear HV in group 1 increased significantly in comparison with the pretreatment period (from 5.7 +/- 2.0 to 10.1 +/- 1.7 cm/year; p < 0.001). Increased HV was sustained during the 2nd and 3rd year of treatment and was significantly higher than at baseline. A similar growth pattern was seen during the 3 years of growth hormone (GH) treatment in group 2. The mean height SDS for chronological age increased by 2.0 +/- 0.7 in the 2 groups after 3 years of treatment. Bone age remained retarded but increased with a mean of almost 4 years after 3 years of treatment in both groups. Even at a dose that is three times the replacement dose, treatment with recombinant human GH was well tolerated. From these results, we conclude that recombinant human GH treatment over 3 years can induce sustained catch-up growth in young children with severe short stature and a history of IUGR. PMID- 9730669 TI - Long-term consequences of intrauterine growth retardation. AB - Recent studies in Europe, North America and the developing world have shown that low birth weight or other indices of abnormal fetal growth in babies born at term are linked with a higher prevalence of raised blood pressure, non-insulin dependent diabetes and cardiovascular disease in late adult life. These findings have led to the 'fetal origins' hypothesis which proposes that fetal adaptations to an adverse intrauterine environment programme persistent physiological and metabolic changes which predispose to these diseases. The mechanisms are unknown, but evidence from animal studies and preliminary evidence in humans suggest that impaired fetal nutrient supply permanently alters neuroendocrine development in the offspring resulting in long-term changes in the set point of adrenocortical and sympathoadrenal hormonal activity. PMID- 9730670 TI - Genetics of Silver-Russell syndrome. AB - The Silver-Russell syndrome (SRS) is generally sporadic, but with sufficient reported cases of dominant and recessive patterns of inheritance to suggest a genetic cause in some cases, at least. No consistent cytogenetic abnormalities have been found although some features of the syndrome have been reported to be associated with structural abnormalities of distal 15q. More recently it has been shown that about 10% of SRS patients have maternal uniparental disomy of chromosome 7 which suggests the presence of a maternally imprinted gene on chromosome 7 that is associated with SRS. In the majority of patients with normal biparental inheritance of chromosome 7 the same gene could be involved if the paternal copy were deleted or mutated so that it is disabled and the maternal copy is silent because of the imprinting. PMID- 9730671 TI - Growth hormone treatment of Russell-Silver syndrome. AB - Russell-Silver syndrome represents a special group of children with intrauterine growth retardation (IUGR) who do not experience catch-up growth and have characteristic dysmorphic features. They also have characteristics of abnormal growth hormone pulsatility, absence of catch-down growth after growth hormone therapy and inappropriate advancement of bone age during the middle childhood years. Data from children with Russell-Silver syndrome should certainly be analysed as a separate group from short children due to nondysmorphic IUGR. Initial data suggests that final height outcome will be improved by using pharmacological doses of biosynthetic human growth hormone. Indeed, the recent data supports the hypothesis of Blizzard's group in 1974 that if growth hormone became available in sufficient quantities, then final height could be altered in IUGR children. In addition, the early recognition and treatment of spontaneous nocturnal hypoglycaemia may well improve the educational achievement of such children. PMID- 9730672 TI - The growth hormone cascade: progress and long-term results of growth hormone treatment in growth hormone deficiency. AB - The growth hormone (GH) cascade and the remarkable advances over the past four decades in our knowledge of its components are considered. It is now over 40 years since human pituitary GH (pit-hGH) was purified and the first GH-deficient patient, a 17-year-old male, was successfully treated with pit-hGH. However, the shortage of pit-hGH limited its use and the dose, the biopotency of preparations varied, strict criteria of GH deficiency (GHD) were used for patient selection including peak plasma immunoreactive GH levels after provocative stimuli of <3.5 5 ng/ml, treatment was not infrequently interrupted, the mean age for initiating treatment was often late in childhood (12-13 years) and the growth deficiency severe (height -4 to -6 SDS), and finally pit-hGH therapy was often discontinued when girls attained a height of 5' and boys 5'5". Nonetheless, the effects of pit hGH were dramatic; the final height SDS increased in isolated GHD to about -2 SDS in boys and -2.5 to -3.0 SDS in girls, and in multiple pituitary hormone deficiencies to between -1 and -2 SDS. Between 1962 and 1985 when the Creutzfeldt Jakob disease crisis struck, the number of GH-deficient patients treated with pit hGH increased from about 150 to over 3,000. The advent of biosynthetic GH (rhGH) and its availability to treat large numbers of idiopathic GH-deficient children (the minimum prevalence rate of which in the USA and UK is between 1 in 3,400 and 4,000) dramatically changed this picture in 1985. It is estimated that more than 60,000 patients have been or are now on treatment. With rhGH treatment the attained mean adult height SDS is now about -1.0, and in our experience with the treatment of patients under 4 years of age, final height may exceed the target height. It is now recognized that (a) the replacement dose of rhGH ranges from 0.175 to 0.35 mg/kg/week and should be individualized; (b) dividing this dose into 6 or 7 daily subcutaneous injections is more effective than giving the same total dose in three weekly portions, and (c) final height correlates significantly with pretreatment chronologic age, height SDS and predicted adult height, duration of therapy, birth length, in some studies height SDS and age at start of puberty, weight, and serum GHBP (an indicator of GH receptor mass). Early recognition of GHD is essential for an optimal height outcome. rhGH treatment should not be delayed in children with documented GHD; the greater the height deficit, the lower the probability that target height will be reached. GHD needs to be detected earlier in children with organic hypopituitarism whether due to a developmental defect, neoplasm, radiation, head trauma, or a CNS infection. Early rhGH therapy in neonatal hypopituitarism has resulted in excellent growth responses. As the height prognosis in isolated GHD is not as good (especially in girls) as in GHD associated with gonadotropin deficiency, the use of LHRH agonists to delay puberty or potent aromatase inhibitors to delay skeletal maturation should be considered in selected patients with isolated GHD. When the growth response to rhGH is less than predicted, one must consider: (a) poor compliance; (b) improper preparation of rhGH for administration or faulty injection techniques; (c) the timing of administration; (d) the dose of glucocorticoid in the ACTH-deficient patient; (e) occult hypothyroidism; (f) inadequate nutrition; (g) a chronic illness; (h) neutralizing antibodies to rhGH, and (i) the wrong diagnosis. The major cause of mortality (unrelated to Creutzfeldt-Jakob disease or a CNS neoplasm) is adrenal crisis and hypoglycemia in children with both GH and ACTH deficiency. Major adverse effects of rhGH treatment in children are uncommon and include idiopathic intracranial hypertension, slipped capital femoral epiphysis, and acute pancreatitis. The rhGH is not an added risk for leukemia in the US and Europe in the absence of coexisting risk factors, nor is there a higher risk of recurrence of b PMID- 9730674 TI - Growth hormone therapy in patients with Turner syndrome. AB - This article summarizes the published data on final height after growth-promoting therapy in Turner syndrome. Using growth hormone (GH) doses ranging between 0.5 and 1.2 IU/kg/week, final height after therapy is improved by 1.5-9.3 cm [final height after therapy vs. projected adult height (PAH)] within various studies. There is no obvious GH dose-response relationship, but a better estimated benefit from therapy seems to result in those studies that combined even rather low GH doses with the anabolic steroid oxandrolone. It is not possible to retrospectively define an optimal treatment regime out of the various published data due to different GH doses, age and dosage of estrogen replacement therapy and the variable methods for calculating the benefit from therapy. It seems to be essential to start estrogens at a safe bone age (> 13 years) in very low doses. Higher GH doses (up to 2.0 IU/kg/week) led to a better growth response during the first years of therapy but data on final height are still to be awaited. PMID- 9730673 TI - Management of recombinant human growth hormone therapy at puberty. AB - Treatment modalities for growth hormone (GH) substitution in GH-deficient children at puberty is still a matter of debate. Although circulating GH levels increase during puberty, it has not been proven that the increase of exogenous GH is necessary for a normal pubertal growth spurt. Increasing GH levels may thus well be the consequence and not the cause of the pubertal growth spurt. A permissive role of GH for pubertal growth has been hypothesized. Some effects of exogenous GH may even be detrimental to pubertal growth; specifically, puberty seems to be shortened by GH administration. Further aspects of the physiology of GH secretion and the poor imitation by current replacement schemes are discussed. Only randomized, prospective studies will allow to define the optimal dose of GH at puberty. As long as these studies are missing, a pragmatic approach is an individualized, minimalistic GH substitution scheme under close surveillance. GH dose at puberty should be kept at prepubertal levels, but must be increased once height velocity drops below the 50th percentile. This widely used approach ensures unnecessary, costly and potentially harmful overdosages of GH. PMID- 9730675 TI - High-dose growth hormone therapy in idiopathic short stature. AB - This article reviews the available data on the effect of growth hormone (GH) administration on the growth of children with idiopathic short stature. The 1st year growth response appears clearly dose dependent, if the mean values from various clinical trials are combined. Dose dependency has also been shown in prospective trials. The dose dependency of the long-term growth response was less convincing in a large prospective study. The effect on final height cannot be proven with certainty in uncontrolled studies. If all available results from clinical trials are combined, a positive correlation between response and dose appears likely. PMID- 9730676 TI - The effect of different growth hormone administration frequencies on growth in growth hormone-deficient patients. AB - In two different groups of clinically prepubertal children (bone age 30 mg/24 h) urinary albumin excretion rate (UAER) and the prevalence of albuminuria at the 9-year examination was 35% (29 of 83). During the follow-up period, systolic blood pressure (p = 0.044), prevalence of hypertension (p < 0.01), and fasting blood glucose levels (p < 0.01) increased, while high-density lipoprotein cholesterol (p < 0.01) decreased. The declines of GFR during the follow-up period were 8.5, 14.1, and 16.3% within genotype groups of II, ID, and DD, respectively (p values for decreases: NS for II, <0.001 for ID, and <0.001 for DD). Patients with the DD genotype tended to have a steeper decrease of GFR, but the change was not statistically significant between the genotype groups. The increases of UAER during the follow-up period were 35.1, 8.3, and 122.4% within genotype groups of II, ID, and DD, respectively, but p values for all increases were not significant. Parallel to GFR, patients with the DD genotype tended to have a steeper increase of UAER, but the change was not statistically significant between the genotype groups. There were no differences in the ACE genotype distribution and allele frequencies between the patients with or without albuminuria either at follow-up or in cross-sectional settings. In conclusion, this 9-year follow-up study does not support the hypothesis that the ACE I/D polymorphism is a major genetic marker of diabetic nephropathy in NIDDM patients. PMID- 9730699 TI - Tissue-specific expression of human angiotensin II AT1 and AT2 receptors and cellular localization of subtype mRNAs in adult human renal cortex using in situ hybridization. AB - All studies analyzing the localization of angiotensin II (Ang II) receptors in the human kidney have been performed at the protein level using 125I-Ang II as a probe. In this study, cellular localizations of Ang II type l (AT1-R) and type 2 (AT2-R) receptor mRNAs in the adult human renal cortex were examined for the first time using in situ hybridization, and their expression patterns determined by RNase protection assay were compared with those in other human tissues. In the human renal cortex obtained from tumor-free portions in renal cell carcinoma, AT1 R mRNA levels were about 8- to 10-fold higher than AT2-R mRNA levels. Human liver and aorta predominantly expressed AT1-R mRNA, while human right atrium contained both AT1-R and AT2-R mRNAs. Ligand-binding assays revealed that the total Ang II receptor number in the human renal cortex was 16.0 +/- 3.3 fmol/mg protein, similar to that in liver (17.7 +/- 5. 8) but significantly higher than in right atrium (11.6 +/- 3.2) and aorta (5.6 +/- 2.7). Relative distribution ratios of AT1-R and AT2-R numbers in the renal cortex and right atrium were 82/17 and 56/42%, respectively. In situ hybridization study indicated that strongest AT1-R mRNA signals were located in interlobular arteries and tubulointerstitial fibrous regions surrounding interlobular arteries and glomeruli, followed in decreasing order by glomeruli and cortical tubules. Expression of AT2-R mRNA was highly localized in interlobular arteries. Cells present in tubulointerstitial regions were positive for vimentin and collagen type 1, indicating that the majority of the cells present in the regions are fibroblasts. Presence of strong AT1-R mRNA signals in the tubulointerstitial fibrous regions surrounding arteries and glomeruli and the expression of AT2-R mRNA in the interlobular artery were the first evidence, suggesting a pharmacological framework for the differential effects of Ang II receptor subtype mediated renal function in the adult human kidney. PMID- 9730700 TI - Serum leptin concentrations in patients on hemodialysis. AB - Serum leptin concentrations in normal humans have been reported to correlate with the body mass index (BMI) as well as with the body fat mass. In this study, we measured serum leptin concentrations in 107 patients on hemodialysis, 30 of whom had diabetes mellitus as the cause, and examined the clinical significance. Furthermore, we evaluated the effects of high-flux dialysis membranes on serum leptin levels. Serum leptin concentrations had a linear correlation with BMI as well as with the percentage of body fat in patients on hemodialysis. The serum leptin concentrations showed a positive correlation with the serum concentrations of total cholesterol, low-density lipoprotein cholesterol, and triglyceride, the body weight, the BMI, and the percentage of body fat. The serum leptin levels were not different between the diabetic and the nondiabetic groups. The serum leptin levels in the nondiabetic group were nearly fourfold higher in women than in men. We investigated the differences in the rate of reduction in serum leptin after dialysis with polysulfone membrane dialyzers (PS-N and PS-UW) in comparison with a cellulose membrane dialyzer (AM-SD), and as a result, we found that the polysulfone membrane dialyzers removed serum leptin, while the cellulose membrane dialyzer did not. We conclude that in patients on hemodialysis, the serum leptin concentration is a valuable clinical marker of the body fat content and may also contribute to the evaluation of hyperlipidemia. PMID- 9730701 TI - Stimulation of hepatocyte growth factor in human acute renal failure. AB - Hepatocyte growth factor (HGF) is a potent mitogen for tubular cells. Experimental injury to the kidney is associated with HGF release both locally and by distant organs stimulated by circulating 'injurins'. In this study, the serum HGF concentration was measured in patients with acute renal failure (ARF). Normal subjects and chronic renal failure patients either not on dialysis or on regular dialysis treatment served as controls. Human mesangial cells were incubated with sera from ARF patients and controls. The serum HGF concentration was strikingly increased in ARF patients (478 +/- 68 ng/dl) and was normal in chronic renal failure patients not on dialysis (20 +/- 3 ng/dl) and in those on regular dialysis treatment (25 +/- 3 ng/dl). Serum of ARF patients strongly stimulated HGF release from mesangial cells (1,384 +/- 55 ng/ml) in comparison with normal serum (67 +/- 10 ng/ml). These results indicate that in ARF HGF participates in tubular repair both as an endocrine factor, released in the circulation, and as a paracrine substance, diffusing to the tubules from the mesangium. PMID- 9730702 TI - Effects of uremic plasma on alpha- and beta-adrenoceptor subtypes. AB - Responsiveness of adrenergic receptors is decreased in patients on maintenance hemodialysis. In this study we investigated whether uremic plasma might affect adrenergic receptors. For this purpose we determined the effects of uremic plasma obtained from 10 patients on hemodialysis treatment (mean age 61 +/- 3 years, dialysis frequency 3 x 4 h/week, duration of treatment 3 +/- 1 years) before and at the end of the 4-hour dialysis treatment on binding of radioligands to beta1- and beta2- as well as alpha1- and alpha2-adrenoceptors. Plasma from 6 healthy volunteers served as control; the plasmas were studied in three dilutions: undiluted, 1:1 (v/v) and 1:4 (v/v) with saline diluted. Plasma from healthy control did not significantly affect the number of beta1- and beta2- or alpha1- and alpha2-adrenoceptors. On the other hand, uremic plasma significantly decreased the number of beta1- and beta2-adrenoceptors; this inhibitory effect was also observed when plasma obtained at the end of the 4-hour dialysis treatment was investigated. On the other hand, uremic plasma did not significantly decrease the number of alpha1- and alpha2-adrenoceptors. We conclude that in patients on maintenance hemodialysis, the presence of inhibitory substance(s) in uremic plasma could be - at least partly - responsible for the beta-adrenoceptor hyporesponsiveness; the mechanism leading ot alpha-adrenoceptor hyporesponsiveness, however, remains to be elucidated. PMID- 9730703 TI - Interferon therapy for chronic hepatitis C in hemodialysis patients: increased serum levels of interferon. AB - BACKGROUND/AIMS: The efficacy and side effects of interferon (IFN) therapy have not been well clarified in hemodialysis patients with chronic hepatitis C. METHODS: In 6 of 9 hemodialysis patients with chronic hepatitis C, 3 million units (MU) or 6 MU of recombinant IFN-alpha2b or natural IFN-alpha were administered intramuscularly daily for the first 2 weeks, followed by three times a week for 22 weeks. In the remaining 3 patients, 3 MU of IFN-alpha2b were given three times a week for 24 weeks. Serum concentrations of IFN-alpha2b were measured sequentially after the injection of interferon. Responders were defined as the patients with normal serum aminotransferase and negative serum HCV RNA 6 months after the cessation of IFN therapy. RESULTS: Three of the 6 patients who were administered IFN daily in the first 2 weeks were responders, while the other 3 withdrew from the therapy due to serious adverse events such as depression, loss of consciousness and persistence of high-grade fever. Serious adverse events were not observed in the 3 patients without daily administration. Half-lives of IFN-alpha2b in hemodialysis patients were significantly longer than those in nonuremic patients (10.0 vs. 6.0 h, p < 0.05). Moreover, the areas under the serum concentration curve of the hemodialysis patients were significantly larger than those of nonuremic patients (756 +/- 223 vs. 324 +/- 223 IU.h/ml, p < 0.05), despite the fact that the dose of IFN-alpha administered to hemodialysis patients was half that administered to nonuremic patients. CONCLUSIONS: In hemodialysis patients with chronic hepatitis C, pharmacokinetic parameters of IFN may be different from those in nonuremic patients, and daily or high-dose administration of IFN may lead to serious adverse events in those patients. PMID- 9730704 TI - Improvement of uremic autonomic dysfunction after renal transplantation: a heart rate variability study. AB - Autonomic dysfunction in hemodialysis patients is one of the components of uremic neuropathy. In this prospective study, we investigated the effect of renal transplantation on uremic autonomic dysfunction with long-term time-domain and frequency-domain heart rate variability. Fourteen hemodialysis patients (10 male, 4 female; mean age 33 +/- 11 (range 16-50) years) were examined before and at the early after transplantation period (mean 4.6 +/- 1.5 (range 3-7. 5) months). The mean time spent on hemodialysis was 16.7 +/- 15.6 (range 6-65) months. In time domain analysis, significant increases in all parameters except pNN50 (SD, SDANN, SDNN, rMSSD) were observed after renal transplantation (p < 0.01). In frequency domain analysis, low-frequency (LF) (0.04-0.15 Hz) and high-frequency (HF) (0.15 0.40 Hz) spectral power were found to be significantly increased after renal transplantation (4.54 +/- 1.04 vs. 12.58 +/- 8. 69 for LF (p = 0.005), 2.80 +/- 1.0 vs. 6.50 +/- 3.55 for HF (p = 0. 005)), but the LF/HF ratio was not different from a pretransplant period (1.71 +/- 0.349 vs. 1.85 +/- 0.49, p = 0.26). It was concluded that autonomic dysfunction in hemodialysis patients is reversible and renal transplantation reverses the sympathetic and parasympathetic autonomic dysfunction simultaneously and at a relatively early stage. PMID- 9730705 TI - Relapsing tinea capitis by Microsporum canis in an adult female renal transplant recipient. AB - Scalp ringworm is very uncommon in adults. The occurrence and the atypical clinical course of this unusual dermatophytosis in a female renal transplant recipient are described. Furthermore, the prevalence and the clinical features of superficial fungal infection in renal transplant recipients are reviewed. As immunosuppression enhances the risk of antifungal therapy failure, more prolonged treatment and careful follow-up are necessary to obtain complete recovery from any dermatophytosis in renal transplant recipients. PMID- 9730706 TI - Reversible neuropathy in chronic renal failure. AB - Chronic renal failure is associated with distal symmetrical axonal neuropathy. We report a 50-year-old man who suffered from chronic renal failure due to benign enlargement of the prostate. He presented with fever and rapidly developing pure motor neuropathy. The nerve conduction velocity was slow and the F response delayed, suggestive of demyelinating neuropathy. A sural nerve biopsy specimen was also consistent with demyelination and axonal changes. This patient improved significantly following intermittent catheterization and a short course of norfloxacin. Renal failure may be associated with reversible motor neuropathy. PMID- 9730707 TI - Life-threatening hematuria from an arteriovenous fistula complicating an open renal biopsy. AB - Arteriovenous fistulae and pseudoaneurysms are not rare after renal biopsy. The majority of these lesions (80%) are asymptomatic or show only transient symptoms. We present here a patient who developed life-threatening hematuria following an open renal biopsy. Arteriovenous fistula and pseudoaneurysm were detected in the biopsied kidney by color-coded Doppler sonography, confirmed by angiography, and the fistula was sealed by superselective arterial embolization with metallic coils. Color-coded Doppler sonography successfully detects the majority of arteriovenous fistulae after renal biopsy, and selective arterial embolization obviates the need for surgical intervention in most cases. PMID- 9730708 TI - Atraumatic loss of a kidney in a patient with alpha1-antitrypsin deficiency. AB - Alpha1-Antitrypsin (alpha1AT) deficiency is characterized by a high risk of developing pulmonary emphysema and liver disease. Arteriopathy may be associated with alpha1AT deficiency. We present a patient with only one kidney and alpha1AT deficiency, but only mild pulmonary symptoms and no signs of liver disease, and atraumatic loss of the remaining kidney. Histological examination of the renal artery showed desposits of a mucoid ground substance in the arterial media, as has been demonstrated in other patients with alpha1AT deficiency. After dialysis treatment for 15 months, the patient underwent an uneventful kidney transplantation. The case draws attention to the awareness of complications of alpha1AT deficiency even in cases with only mild pulmonary manifestations of the disorder. To our knowledge, atraumatic loss of a kidney in a patient with alpha1AT deficiency has not previously been reported. PMID- 9730709 TI - 99mTc-labelled red blood cell scintigraphy for localization of gastrointestinal bleeding in chronic renal failure. AB - Angiodysplasia (AD) may be the source of bleeding in patients with gastrointestinal hemorrhage, with special occurrence in the elderly population and in patients with chronic renal failure (CRF). Although several techniques have been tested for its diagnosis, the gold standard is not well defined yet. We analyze the usefulness of 99mTc-labelled red blood cell (99mTc RBC) scintigraphy in the localization of bleeding from AD lesions in a cohort of 21 patients. Other investigative methods include fibrocolonoscopy examination, angiography, or diagnostic laparotomy. Group A (AD and CRF): 11 patients. Group B (AD without CRF): 10 patients. 99mTc RBC scintigraphy showed 88.9% sensitivity and specificity in group A, while in group B it had 100% sensitivity and specificity. Arteriography showed 100% sensitivity and specificity. On the contrary, fibrocolonoscopy had a very low sensitivity (30%). Our results suggest that 99mTc RBC scintigraphy may be the preferred diagnostic tool for AD, especially in patients with CRF, in whom arteriography may accelerate the decline of renal function. PMID- 9730711 TI - Prevalence of hepatitis E virus antibody in hemodialysis patients. PMID- 9730710 TI - Paper chromatography prior to cortisol RIA allows for accurate use of the dexamethasone suppression test in chronic renal failure. AB - The assessment of the hypothalamic-pituitary-adrenal axis in patients with chronic renal failure (CRF) on hemodialysis is often hampered by abnormal responses to the standard 1-mg dexamethasone suppression test. Various mechanisms have been proposed to explain this lack of suppressibility. The present study was designed to look into the mechanisms possible for these findings in patients with CRF. We studied 6 patients with CRF on hemodialysis and 5 healthy subjects utilizing the 1-mg dexamethasone suppression test as well as the 50-mg hydrocortisone suppression test. Samples were assayed for dexamethasone, adrenocorticotropic hormone, corticosterone, and cortisol by both direct radioimmunoassay (RIA) and RIA after paper chromatography. Utilizing the direct cortisol RIA, 4 of 6 patients with CRF exhibited blunted dexamethasone suppression, while all 6 patients showed normal suppressibility after dexamethasone when cortisol was measured after paper chromatography. In contrast, all controls showed normal suppressibility regardless of the cortisol assay procedure used. The hydrocortisone suppression test was unreliable in the setting of CRF. Mean dexamethasone levels were similar in both groups. Plasma adrenocorticotropic hormone levels were significantly higher in the CRF patients, possibly indicative of an underlying hypothalamic-pituitary-adrenal axis abnormality. Abnormalities in dexamethasone suppression testing in patients with CRF may be explained by the overestimation of cortisol levels by direct RIA rather than by alteration of dexamethasone absorption or metabolism. Measurement of cortisol after paper chromatography is superior to direct RIA of cortisol in patients with CRF. PMID- 9730713 TI - Plasma carnitine levels in patients undergoing hemodialysis. PMID- 9730712 TI - Prolonged cannulation of the femoral vein is a safe method of temporary vascular access for hemodialysis. PMID- 9730714 TI - Vitamin D3 withdrawal in hemodialysis patients with adynamic bone disease. PMID- 9730715 TI - Size limitations to adequacy of small-solute clearance in peritoneal dialysis: body surface area as the size indicator. PMID- 9730716 TI - Role of chlamydia pneumoniae (TWAR) in IgA nephropathy. PMID- 9730717 TI - Lack of serologic evidence for an association between Hantavirus infection and end-stage renal disease. PMID- 9730718 TI - Hemostasis, platelet functions, serotonin and serum lipids during omega-3 fatty acid treatment in patients with glomerulonephritis. PMID- 9730719 TI - Is there an association between glomerular density and birth weight in healthy humans. PMID- 9730720 TI - Detection of urinary MCP-1 in patients with diabetic nephropathy. PMID- 9730721 TI - Newly diagnosed insulin-dependent diabetes mellitus with rapid severe thickening of the glomerular basement membrane. PMID- 9730722 TI - Xanthogranulomatous pyelonephritis in a child with cystinuria. PMID- 9730724 TI - Focal segmental glomerulosclerosis and Turner syndrome. PMID- 9730723 TI - Unusual case of membranous nephropathy exhibiting type III collagen microfibrils. PMID- 9730725 TI - Hemolytic uremic syndrome associated with beta-interferon therapy for chronic hepatitis C. PMID- 9730726 TI - Simplified peritoneal equilibration test in CAPD. PMID- 9730727 TI - A case of membranous nephropathy associated with psoriasis vulgaris. PMID- 9730728 TI - Development of eclampsia and end-stage renal disease in a case of Sickle cell disease. PMID- 9730729 TI - Alport syndrome with a peculiar pattern of distribution of the alpha3-alpha6 chains of type IV collagen in renal basement membrane. PMID- 9730730 TI - Gastroduodenal bleeding after discontinuation of H2-receptor antagonists in hemodialysis patients. PMID- 9730731 TI - Effectiveness of intravenous and subcutaneous calcitriol in the treatment of secondary hyperparathyroidism. PMID- 9730732 TI - Cyclosporine induces apoptosis of mouse terminal proximal straight tubule cells. PMID- 9730734 TI - High-frequency ultrasound biomicroscopy versus ultrasound and optical pachymetry for the measurement of corneal thickness. AB - The aim of this study was to correlate central corneal thickness measurements obtained using optical as well as ultrasound pachymeters and high-frequency ultrasound biomicroscopy (UBM), assuming UBM as reference. Each measurement was performed on 32 patients (60 eyes) by three observers unaware of the other's results, on 3 different days. Pearson correlation was used. A strong correlation was found between UBM and ultrasound pachymetry measurements (r=0.858), a weaker one between UBM and optical pachymetry (r=0.506). Optical versus ultrasound measurements were poorly correlated (r=0.540). Each correlation was statistically significant (p<0.001). UBM can be used as an accurate and reproducible method for determining corneal thickness. PMID- 9730733 TI - Renal failure due to IgA (Berger) nephropathy with inflammatory demyelinating polyradiculoneuropathy. PMID- 9730735 TI - Ultrasound biomicroscopy in the assessment of secondary glaucoma after vitreoretinal surgery and silicone oil injection. AB - This retrospective study was undertaken to evaluate the utility of Ultrasound Biomicroscopy (UBM) in the assessment of the anterior chamber in patients affected by silicone oil-related glaucoma after vitreoretinal surgery. UBM showed high reflectivity of angular structure, anterior chamber alterations and a fairly trophic ciliary body. PMID- 9730736 TI - Ultrasound biomicroscopy in the assessment of penetrating or blunt anterior chamber trauma. AB - Ultrasound biomicroscopy (UBM) was employed in the assessment of the anterior chamber in patients affected by penetrating and blunt traumas. UBM detected correctly the extent and the localization of penetrating injuries and the presence of foreign bodies, IOL dislocation, and angular structures or ciliary body damages. PMID- 9730737 TI - The use of ultrasound biomicroscopy in pre-operative evaluation of orbital blepharal neoformations: report of a case. AB - In our study we have taken into consideration the use of ultrasound biomicroscopy (UBM) in a case of orbital-blepharal lymphoma. The advantages of this methodology, which is considered a refinement of B scan ultrasonography, compared with other instruments at present available on the market, have been described. We have tried to show how the instrument for biomicroscopic ultrasonography makes it possible to obtain images of the anterior segment of the eye in every detail; it is, therefore, capable of giving us information once only available by means of examining histopathological sections. Apart from the advantages, in this work we have also described the limits of this methodology, which in order to allow an accurate diagnosis must always be accompanied by the traditional ocular echography. PMID- 9730738 TI - Keratoconus staging with ultrasound biomicroscopy. AB - The diagnosis of keratoconus is based on slip-lamp and keratometric findings, and its classification includes 4 different stages. In this study, we compare the ultrasound biomicroscopy (UBM) findings and the keratoconus index (KI) determined using UBM with the severity of the disease. By means of UBM (Humphrey Instruments system model 840) using a 50-MHz probe, we studied 49 eyes affected with different forms of keratoconus. Using a method previously described, we calculated the KI and compared these data with the findings obtained using videokeratography. The mean thickness at the corneal apex in keratoconic eyes was 0.369 mm, while the mean peripheral corneal thickness was 0.568 mm. The mean KI was 1.449 in keratoconic eyes. The KI increased according to the severity of the disease. The mean central keratometry power was 56.95, while the mean amount of steepening of the inferior cornea compared with that of the superior cornea was 8.16. UBM can be considered a useful tool in the study of keratoconus. Using high frequency ultrasound, it is also possible to obtain reliable measurements of corneal thickness related to the severity of the disease determined by videokeratography. PMID- 9730739 TI - Corneal oedemas: diagnosis and surgical planning with ultrasound biomicroscopy. AB - We used the Humphrey Ultrasound Biomicroscope (UBM System 840 by Zeiss-Humprey Instruments, San Leandro, Calif. USA) to study various corneal oedemas and leucomas. This UBM system, using a high-resolution probe of 50 Mhz, has an axial and lateral resolution of about 50 micrometer. We analysed 36 eyes divided into two groups, 27 (group A) affected by corneal oedema caused by traumatic-mechanic or phlogistic-toxic lesions, 9 (group B) affected by postsurgery oedema. Thanks to its high-resolution power, we could study their pathogenesis and their clinical evolution and so arrange a suitable therapy and perform an accurate follow-up of these pathologies. PMID- 9730740 TI - Ultrasound-biomicroscopic evaluation of filtering blebs after laser suture lysis trabeculectomy. AB - Sometimes in glaucomatous patients treated with trabeculectomy there is no correlation between bleb shape and intra-ocular pressure (IOP). In this study we evaluated the ultrasound biomicroscopy (UBM) features of filtering blebs after laser suture lysis (LSL) trabeculectomy in order to analyse whether its ultrasound-biomicroscopic image can predict the function (IOP). METHODS: The Humphrey ultrasound biomicroscope, using a high-frequency (50-Mhz) probe, provides high-resolution images of filtering blebs. A total of 103 filtering blebs after LSL trabeculectomy were analysed by UBM. Taking into account the characteristics of internal reflectivity and scleral flap, we classified the blebs into three groups (good, fair, poor) that indicate the bleb function and correlated this UBM pattern with the IOP control (good, borderline, failure). RESULTS: There was a statistically significant corelation between the UBM classification of function and the IOP control level. Both well-functioning and failed trabeculectomies could be identified by UBM. The UBM images of eyes with good IOP control are characterized by better visibility of the route under the scleral flap and a low reflectivity inside the bleb. CONCLUSIONS: In accordance with previous studies, we believe that UBM can be a useful method to study and explain the mechanisms of filtering structures and, together with IOP control, to evaluate the bleb function. PMID- 9730741 TI - Extraocular muscle size comparison between normal and myopic eyes using standardized A scan echography. AB - The aim of this study was to establish the range of diameters of extraocular muscles in myopic patients and to obtain the baseline data to follow progression or regression of pathologies involving them. Standardized A scan and B scan ultrasound was used to measure the thickness of straight extraocular muscles at the muscle belly. No statistical differences were seen between right and left eyes. We found no consistent correlation with age or with axial length in the control or in the myopic patients (p>0.05). Diameters in myopic and control eyes were similar. This lack of difference between myopic and control muscle sizes means that we can obtain accurate measurements also from myopic patients. PMID- 9730742 TI - Echobiometric evaluation of the axial length of the eye and intraocular lens calculation in pseudophakic eyes: our experience. AB - Intraocular lens (IOL) power calculation in 46 pseudophakic eyes (extracapsular cataract extraction with IOL in posterior chamber), utilizing a Javal keratometer, a Sonomed A 2000 echobiometer (probe 10 MHz, velocity=1,548 m/s) and the SRK2 formula, although there was a statistically significant reduction of the axial length, both in normal and hyperopic eyes, demonstrated no statistically significant differences of IOL power, when compared to the power previously calculated in the phakic eye. PMID- 9730744 TI - Biometric pre-operative evaluation of ocular axial length in patients subjected to corneal transplantation. AB - In cases of keratoconus and consequent penetrating keratoplasty, the refractive requirements of the patient determine the dimensions of the donor button and the receiving wound size. The pre-operative biometric evaluation of the axial parameters may induce the surgeon to choose an oversized button to obtain the right curvature of the cornea and avoid hypermetropia. In this study we present 20 patients transplanted between 1993 and 1995, 13 female, 7 male, aged between 23 and 45 years. These cases demonstrate the utility of this easy and economic technique. PMID- 9730743 TI - Echobiometry of the ocular layers at the posterior pole. AB - The thickness of the retina, choroid and sclera in relation to macular oedema (ME) can be measured using A scan echography. Ninety diabetic patients (180 eyes) divided into two groups with and without ME, respectively, were studied compared to a third group of 73 healthy controls (146 eyes). Statistical analysis of results showed good specificity (80%) and reasonable sensitivity (70%) of the echographic method in showing the presence of ME confirmed by fluorescein angiography. PMID- 9730745 TI - Echographic evaluation of choroidal melanomas treated with ruthenium-106 plaques: the treatment failures. AB - From July 1990 to October 1995 we treated 45 patients with choroidal melanomas using ruthenium-106 plaques. The population mean age was 59.2+/-10.4 years. In this paper, the authors describe in detail their clinical management and follow up timing. They also report on the echographic pattern change after treatment and the clinical history of some failure cases. PMID- 9730746 TI - Rapidly growing choroidal metastasis: case report. AB - A rapidly growing choroidal metastasis from an occult carcinoma is described. The patient was first seen with a loss of visual acuity. The fundus examination showed a non-pigmented solid mass located at the posterior pole. Clinical history, echographic characteristics and tumour growth kinetics are described in detail. PMID- 9730747 TI - Recurrent eyelid basal cell carcinoma with sclerochoroidal infiltration: echographic findings. AB - In an 82-year-old woman with a recurrent right lid basal cell carcinoma infiltrating the upper temporal orbit, a solid yellowish lesion was found ophthalmoscopically in the superotemporal periphery of the right eye. On standardized echography examination, the 2.5-mm elevated subretinal lesion was medium to high reflective with irregular structure, whereas the adjacent orbital mass was medium to low reflective and the sclera was thinned with at least one passage between the orbital and ocular lesions. To our knowledge, this is the first echographic report of sclerochoroidal infiltration from a lid basal cell carcinoma. PMID- 9730748 TI - Ultrasonographic features and patterns of regression after conservative treatment of retinoblastoma. AB - The authors analyze the echographic features relative to some cases of retinoblastoma treated with conservative therapy in order to monitor the regression of the tumor consequent to the treatment. PMID- 9730749 TI - Sensitivity and specificity of ultrasonography, fluorescein videoangiography, indocyanine green videoangiography, magnetic resonance and radioimmunoscintigraphy in the diagnosis of primary choroidal malignant melanoma. AB - The prognosis of primary choroidal malignant melanoma (PCMM) is fatal if no early reliable diagnosis is performed. Any incisional biopsy is impossible, and diagnosis is only based on instrumental examinations. The purpose of this study is to evaluate the reliability of ultrasonography, fluorescein videoangiography (FV), indocyanine green videoangiography (ICGV), magnetic resonance (MR) and radioimmunoscintigraphy (RIS) in the diagnosis of PCMM in a series of 12 eyes in which the tumor was suspected. A presumed diagnosis of PCMM was made when a positive result was obtained with 3 or more methods. The presumed diagnosis was then compared with histological findings (true value) in 4 enucleated eyes. The sensitivity and specificity of every single method were evaluated comparing its results with the final presumed diagnosis and with the histological findings. Sensitivity and specificity of every single method have been expressed as percentage of correspondence with the presumed diagnosis. Sensitivity was 100% for ultrasonography, MR, FV, ICGV and 67% for RIS. Specificity was 92% for ultrasonography, 87% for RIS, 83% for MR and 82% for FV and ICGV. This study indicates that the ophthalmologist can obtain a good diagnostic reliability in the case of PCMM using only ultrasonography, FV and ICGV. Besides MR and RIS are important adjunctive methods to ophthalmological investigations for the diagnosis of PCMM. PMID- 9730751 TI - Ultrasonographic follow-up of primary choroidal malignant melanoma after proton beam irradiation therapy. AB - The aim of this study is to evaluate the ultrasonographic aspects of primary choroidal malignant melanoma (PCMM) after proton beam irradiation therapy and the reliability of its ultrasonographic follow-up in a series of 10 patients. All patients were examined with standardized A and B scan ultrasonography before and after treatment with proton beam irradiation therapy. The follow-up was carried out at 1, 3, 5, 8 months and then every 6 months. The mean follow-up was 20 months (range 6-34 months). The thickness of the tumors was demonstrated to be decreased and the internal reflectivity to be enhanced in 9 out of 10 cases from 8 months after treatment till the end of the follow-up period. Histopathology confirmed in 1 eye enucleated 5 months after treatment that medium-low internal reflectivity coresponded with several areas of high cellularity and cellular mitosis. The results of our study indicate the reliability of standardized A and B scan echography in the follow-up of PCMM treated with proton beam irradiation therapy. PMID- 9730750 TI - Ultrasonographic and angiographic follow-up of primary choroidal malignant melanoma after proton beam irradiation therapy. AB - The aim of this study is to evaluate the reliability of the ultrasonographic and angiographic follow-up of primary choroidal malignant melanoma (PCMM) after proton beam irradiation therapy in a series of 14 patients. All patients underwent standardized A and B scan ultrasonography, fluorescein and indocyanine green videoangiography (FV, ICGV) before and after treatment with proton beam irradiation. The follow-up was carried out at 1, 3, 5, 8 months and then every 6 months. The mean follow-up was 26 months (range 6-46 months). The thickness of the tumors was demonstrated to be decreased and the internal reflectivity to be enhanced in 12 out of 14 cases from 8 months after treatment till the end of the follow-up period. In 2 eyes which were enucleated 5 months after treatment, histopathology confirmed that medium-low internal reflectivity corresponded to several areas of high cellularity and cellular mitosis. FV and ICGV were impossible because of opacities of dioptric media in 2 cases. From 8 months after treatment till the end of the follow-up period, in all the examined patients, FV demonstrated large hypofluorescent zones of the lesions which represented the destruction of the vascular tree of the tumor. From 8 months after treatment till the end of the follow-up period, ICGV demonstrated hypofluorescence of the lesions in 8 cases; mild hyperfluorescence and zones of colorant staining were present in late angiograms in 2 cases. The results of our study seem to confirm the reliability of standardized A and B scan ultrasonography in the follow-up of PCMM treated with proton beam irradiation; if ultrasonography is used together with FV and ICGV, the reliability of this combination is surely higher. PMID- 9730752 TI - Melanoma of the choroid above the optic disc: considerations concerning a clinical case. AB - The authors report the case of a patient suffering from melanoma of the choroid above the optic disc. Given the good level of visual acuity and the location and extent of the damage, he was given a radiotherapy treatment with proton beams. At an interval of 6 months from this treatment, the neoformation appeared limited, and no vascular changes due to irradiation were observed. PMID- 9730753 TI - A case of quiescent choroidal melanoma: early echographic signs of reactivation. AB - A medium-sized choroidal melanoma was studied during a 5-year phase of remission. The tumour presented signs of reactivation with changes in the echographic features without a significant increase in size. The echographic signals in the reactivation phase and the echographic evolution after treatment are described. PMID- 9730754 TI - Echographic follow-up in patients who have undergone sclero-uveotumorectomy. AB - A-B scan echography provides accurate information on the location, the nature, the extension of a neoplasm and the involvement of neighbouring tissues as well as on the postsurgical evolution. In this study, we intend to give accurate and useful indications to the surgeon concerning conservative surgery and the follow up after sclero-uveotumorectomy in cases of uveal neoformations. Between January 1988 and June 1996, we examined 46 patients by means of echography before and after sclero-uveotumorectomy. PMID- 9730755 TI - Congenital retinoblastoma: appearance of calcifications during short-term follow up. AB - An echographic follow-up of a case of congenital retinoblastoma was performed by monitoring variations in the tumoral echostructure concomitant with the appearance of microcalcifications. A previously unreported echo-graphic aspect regarding the first phase of observation is described. The calcifications in the tumour were evidenced on the 41st day of the patient's life. PMID- 9730756 TI - Evaluation of the echographic contribution to the parasurgical treatment of melanomas of small dimensions. AB - In this work we underlined the contribution of echography in the parasurgical treatment (photocoagulative) of melanomas of small dimensions. Fourteen patients were examined; some of them were treated with argon laser, others with krypton laser. The echographic examination was performed with the A-B scan technique, naturally accompanied by fluorangiography before and after the operation. An accurate echographic and fluorangiographic follow-up makes it posible to control the evolution of the lesions efficaciously. From this work it is clear how echography gives highly reliable results which allow us, therefore, both to make an adequate therapeutic decision and to evaluate the efficaciousness of the treatment itself. PMID- 9730757 TI - Retinal astrocytoma associated with hypertrophy of the retinal pigment epithelium: clinical report and follow-up. AB - Retinal astrocytoma is a rare and benign tumor of the retina most frequently seen in patients with tuberous sclerosis. We describe the case of a 12-year-old boy with solitary retinal astrocytoma in the right eye associated with hypertrophy of the retinal pigment epithelium in the left eye. In this work the instrumental examinations, the differential diagnosis and the follow-up of 7 years are described. PMID- 9730758 TI - Regression of choroidal metastasis from a carcinoma of the male breast: case report. AB - Carcinoma of the male breast is rare. The author reports a case of metastatic tumor from carcinoma of the male breast in which was evident, after chemotherapy, a regression of a choroidal metastasis and a reduction of the concomitant cerebral and bony tissue metastases. PMID- 9730759 TI - Ultrasonographic follow-up of patients with choroidal melanoma following conservative treatment. AB - The authors assessed the regression of choroidal tumors, following irradiation treatment, by means of B scan sonography (Sonomed B 3000). Thirty-two patients were studied, 12 of whom underwent brachytherapy with 106Ru plaques and 20 of whom were treated with accelerated protons. After a follow-up period of 12 months, the following was observed: reduction of the thickness of the tumor (significantly greater in the tumors which underwent brachytherapy) and morphological and structural changes which consisted in a thinning of the tumor and an increased reflectivity. PMID- 9730760 TI - Conservative treatment of adnexal, epibulbar and intraocular neoplasms by means of electron microbeams: a preliminary study. AB - At present, episcleral plaque brachytherapy and charged particle or photon irradiation are the most commonly employed methods in ocular and adnexal conservative treatments. In the near future, a different therapeutic approach to these malignancies could be represented by a new device based on electron beam emission. The equipment used consists of an electron accelerator originally developed for intraoperative radiotherapy, modified to fit ocular pathologies. Adequate collimators and an enhanced mechanical accuracy are required for this special practice. An operative planning for epibulbar, adnexal and intraocular tumors is described, discussing its rationale for possible applications. Experimental tests using phantoms and Gafchromic(R) films are in progress. As all conservative treatments, the main gaol of this activity is the maintenance of a good visual function. PMID- 9730762 TI - In vitro evaluation of a new echographic contrast agent. AB - BACKGROUND: So far, intravenously injected echographic contrast agents have not been able to overcome the lung circle, and their use was confined to the study of right-sided heart abnormalities. A new agent that is able to pass the pulmonary barrier has been manufactured, and it is currently under investigation in ophthalmology. METHODS: A saccharide-based contrast agent (SHU 454) that is not able to overcome the lung filter has been compared in vitro with a new saccharide based contrast agent (SHU 508 A) that is able to overcome the pulmonary barrier. They have been diluted in saline solution, and the reflectivity of the solution and the eventual sound attenuation have been studied with standardized A scan and a tissue model. RESULTS: The solution with SHU 508 showed a higher reflectivity that lasted longer, without significant sound attenuation. CONCLUSIONS: The long lasting period of the increased reflectivity will allow to better evaluate the normal and pathological vascular network in the eye and orbit, such as the evaluation of the effectiveness of conservative treatment in cases of malignant melanoma with a spread of the indications of standardized echography to other fields of ophthalmology. PMID- 9730761 TI - Orbital metastases: echographic findings our experience. AB - In this paper the authors examine the echographic findings in 20 patients with metastatic and secondary tumors of the orbit observed at the University Eye Clinic of Ferrara (Italy) between January 1985 and August 1995. The aim of this study is to identify some typical echographic characteristics in these forms and to underline the diagnostic role of orbital echography. PMID- 9730763 TI - Echographic study of the vitreoretinal interface in giant retinal tears. AB - Vitreous and vitreoretinal interface changes were evaluated in giant retinal tears in order to plan surgical approach. Vitreal sineresis was evident in patients affected by primary giant retinal tears; in patients affected by retinal disinsertion the vitreous was adherent to the edge of the posterior flap, and in giant retinal tears secondary to IOL implantation, posterior vitreous detachment but no sineresis was displayed. PMID- 9730764 TI - Reliability of standardized echography before vitreoretinal surgery for proliferative diabetic retinopathy. AB - Standardized A- and B-scan echography was performed in 45 patients (68 eyes) affected by proliferative diabetic retinopathy, before vitreoretinal surgery, to evaluate the percentage of agreement between the ultrasonographic data and the surgical findings. The agreement was quite good for tractional retinal detachment (93%) and for posterior vitreous detachment (91%). The main cause of failure was the presence of vitreoschisis, while preretinal and vitreal membranes can easily be detected. PMID- 9730765 TI - Neoplasia-like echographic evidence in retinopathies of several aetiologies: a case report. AB - In ocular retinal pathology, several aetiological entities showing various clinical and ophthalmoscopic pictures can simulate a tumour. The contribution of echography to the diagnostic research in ophthalmology is really important and the good knowledge of the difficulties and echographic limits is essential. We specifically report a tubercular granuloma clinically and echographically simulating a ciliary-body melanoma. PMID- 9730766 TI - Bilateral occlusion of the central retinal artery in a homocystinuric patient: the role of echography. AB - The authors describe a case presented by a homocystinuric 26-year-old male patient with bilateral occlusion of the central retinal artery. Echography carried out the same day of the occlusion and the following days identified and monitored a mass located in the orbit of the OD. Characteristics and dimension of the mass were confirmed and measured by a CT scan and NMR examination. The orbital mass of the OD appeared to be a swelling of the central retinal artery with echo-refracting opacities, which, in light of recent studies, can be interpreted as multiple arterial thrombi, composed predominantly most likely by platelets. Therapeutic possibilities are discussed. PMID- 9730767 TI - Power doppler ultrasonography in ocular and orbital diseases. AB - PURPOSE: The authors present a preliminary report to test the usefulness of a new technique called power Doppler ultrasonography (PD) in the study of orbital vessels. MATERIAL AND METHODS: The ophthalmic artery and vein as well as the central retinal artery and vein have been examined in 10 patients with a General Electric Logiq 5000 that allows the examination with both color Doppler ultrasonography (CD) and PD. RESULTS: PD does not alias, is relatively angle independent and is able to better detect the pathways of these vessels. CONCLUSIONS: PD is a new promising technique that may be superior to CD in detecting orbital and ocular vessels. PMID- 9730768 TI - Standardized echography, pattern electroretinography and visual-evoked potential and automated perimetry in the early diagnosis of Graves' neuropathy. AB - Twenty-four patients (47 eyes) affected by Graves' disease were enrolled to evaluate the role of standardized echography, pattern electroretinogram (P-ERG), visual evoked potentials (P-VEPs) and automated perimetry in the early diagnosis of the compressive optic neuropathy (CON). The P-ERG amplitude reduction was the most sensitive parameter to demonstrate an early impairment of the optic nerve (ON) function. We found a significant negative correlation between the ON diameter and the P-ERG amplitude. VEPs responses were also altered, but their ability in detecting an early ON damage was less sensitive and specific than P ERG. The visual field damage was often aspecific and delayed with respect to electrophysiological alterations. PMID- 9730769 TI - Graves' disease: measurement of the extraocular muscle thickness with the echobiometer. AB - The authors measured extraocular muscle thickness in normal subjects and in patients affected by Graves' disease, using a Sonomed A-2000 echobiometer (probe with 10-MHz frequency); Hertel's exophthalmometry was also performed. Statistically significant differences in muscle thickness between normals and patients were found. This technique seems to be sufficiently useful and reliable in extraocular thickness evaluation, showing data similar to those of the recent literature. PMID- 9730770 TI - Alterations of the internal reflectivity of extra-ocular muscles associated with several clinical stages of Graves' ophthalmopathy. AB - In the period between February 1992 and June 1996, the authors performed extra ocular muscle echobiometry on 163 patients with Graves' disease. The aim of this study is the evaluation of changes occurring in several clinical stages. The authors noted an increase in internal muscular reflectivity in the advanced phases and an irregular structure in the early stages. Controversial results in the literature warrant mastery of the method and further research. PMID- 9730772 TI - Immersion technique in anterior-segment examination. AB - Ultrabiomicroscopy, using equipment with high-frequency transducers, is a very simple method for the study of superficial ocular structures. Nevertheless, these equipment are very expensive and not commonly used. The most widespread and used instrumentations are still provided with lower-frequency probes. These are sufficient to show the anterior segment of the eye. In this work we underline the diagnostic importance of this method and, in some cases, the therapeutic role. PMID- 9730771 TI - Posterior scleritis: ultrasound findings in two cases. AB - Two cases of posterior inflammatory scleritis are described. The authors show the echographic findings, pointing out the helpfulness of diagnostic ultrasound and the crucial role in monitoring these diseases with A scan examinations to obtain clear results of therapy, even if subjective symptoms disappear. PMID- 9730773 TI - Pediatric ultrasound: a personal experience during the period 1991-1994. AB - The ultrasound examination of the eye with A and B scan has become one of the principal diagnostic methods for detecting ocular abnormalities in children because it is a non invasive technique and can be performed without resorting to general anesthesia. The authors report the most unusual and interesting cases from 1,226 examinations performed in the Department of Ophthalmology, Istituto G. Gaslini, Genova, Italy, between January 1991 and December 1994. PMID- 9730774 TI - Effect of desmethyl tirilazad, dizocilpine maleate and nimodipine on brain nitric oxide synthase activity and cyclic guanosine monophosphate during cerebral ischemia in rats. AB - The present study was designed to evaluate the effects of pretreatment with a combination of desmethyl tirilazad (21-aminosteroid) plus dizocilpine maleate (N methyl-D-aspartate receptor antagonist) and nimodipine (calcium channel antagonist) on constitutive nitric oxide synthase (cNOS) activity and cyclic guanosine monophosphate (cGMP) levels in brain homogenates of rats subjected to global cerebral transient ischemia induced by bilateral clamping of the carotids for 30 min and reduction of arterial pressure (to 50-60 mm Hg) by intravenous infusion of trimethaphan (30 mg/kg). Our results show that cerebral ischemia produced an increase in cNOS activity and cGMP levels in brain homogenates. Pretreatment with desmethyl tirilazad or dizocilpine maleate or nimodipine individually significantly suppressed (p < 0.01) the increase in cNOS activity and cGMP levels induced by cerebral ischemia, which may be related to their neuroprotective effect. Similar results were obtained with pretreatment by a combination of desmethyl tirilazad plus dizocilpine maleate plus nimodipine. PMID- 9730775 TI - Bopindolol: a slowly dissociating antagonist from the beta-adrenoceptors in guinea pig atria. AB - The dissociating and/or residual inhibitory effects of bopindolol from beta adrenoceptors of atria strips pretreated with this drug which was then washed out with buffers on the responses to isoprenaline were determined and compared with those of propranolol, pindolol, atenolol, and the two active metabolites of bopindolol: 18-502 and 20-785. Low concentrations of bopindolol (10(-9) to 10(-8) mol/l) and the active metabolite 18-502 (10(-9) mol/l) produced rightward shifts of the concentration-response curves. On the other hand, high concentrations of bopindolol (10(-7) mol/l) and metabolite 18-502 (10(-8) and 10(-7) mol/l) produced a reduced maximum response by isoprenaline, suggesting that these nonparallel rightward shifts have pD2 values of 7.57 (bopindolol) and 7.67 (18 502), respectively, at 0 min after washout with buffers. Pindolol (10(-7) mol/l) and propranolol (10(-7) and 10(-8) mol/l) also produced a rightward shift of isoprenaline response curves, and these concentration-response curves in guinea pig atria strips pretreated with pindolol (10(-7) mol/l) and propranolol (10(-6) mol/l) recovered to control levels. Neither of these drugs, however, reduced the maximum response by isoprenaline. A high concentration (10(-5) mol/l) of atenolol was required for a rightward shift of the isoprenaline concentration-response curve, and this drug also did not reduce the maximum response. Thus, we conclude that bopindolol and metabolite 18-502 slowly dissociate and act as noncompetitive beta-antagonists rather than easily reversible beta-adrenoceptor antagonists. PMID- 9730776 TI - Contractile and relaxant effects of tetrapentylammonium ions in rat isolated mesenteric artery. AB - Both contractile and relaxant responses to tetrapentylammonium ions (TPA+) were studied in rat isolated mesenteric artery. TPA+ (5-10 micromol/l) caused a sustained increase of muscle tension. The contractile effect of TPA+ (10 micromol/l) was dependent upon the presence of extracellular Ca2+ but independent of the presence of endothelium. TPA+ (10-50 micromol/l) induced biphasic contraction, and the amplitude of peak and sustained tension decreased with increasing TPA+ concentration. TPA+ (100-300 micromol/l) only produced monophasic contraction. TPA+ (50 micromol/l) abolished the transient contraction induced by caffeine (10 mmol/l) or phenylephrine (1 micromol/l) in the absence of extracellular Ca2+. Nifedipine and verapamil concentration-dependently reduced the TPA+-induced contraction with respective IC50 values of 1.34 +/- 0. 24 and 9.46 +/- 1.36 nmol/l, these values were similar to 1.35 +/- 0. 21 and 16.07 +/- 1.71 nmol/l, respectively, for the inhibitory effects of nifedipine and verapamil on the high K+ (60 mmol/l)-induced contraction. TPA+ (>10 micromol/l) concentration-dependently reduced the phenylephrine (1 micromol/l)-, U46619 (30 nmol/l)-, endothelin I (10 nmol/l)- and high K+ (60 mmol/l)-induced sustained tension with respective IC50 values of 53. 7 +/- 9.5, 31.9 +/- 5.3, 30.9 +/- 3.4 and 20.9 +/- 2.8 micromol/l. The present results indicate that TPA+ at low concentrations could contract the arterial smooth muscle probably through promoting Ca2+ influx. At higher concentrations (>20 micromol/l), TPA+ relaxes arterial smooth muscle probably through inhibition of both nifedipine-sensitive Ca+ channels and internal Ca2+ release. TPA+, unlike other quaternary ammonium ions, could therefore act at multiple sites in arterial smooth muscle. PMID- 9730777 TI - Vascular and excretory effects of angiotensin II in the rat isolated perfused kidney: influence of an AT1 and a nonselective AT receptor antagonist. AB - Angiotensin II (AII) is a potent vasoconstrictor which, at physiological plasma concentrations, produces antinatriuresis, whereas high intrarenal concentrations cause natriuresis and diuresis. We examined the effects of a selective AT1 receptor antagonist, losartan, and a nonselective AT receptor antagonist, Sar1Thr8AII, on the response to infusion of AII in the isolated rat kidney perfused at constant pressure with a recirculating modified Krebs-Henseleit buffer. AII increased renal vascular resistance (RVR), glomerular filtration rate (GFR) and urinary volume (UV) and sodium excretion (UNaV) without changing the fractional excretion of water or electrolytes. Thus, changes in GFR can account for the natriuresis/diuresis. Both AII receptor antagonists prevented the increase in RVR. However, losartan was without effect on angiotensin-induced increases in GFR, UV or UNaV, whereas Sar1Thr8 AII also prevented the increases in GFR, UV and UNaV. The angiotensin receptor mediating the increase in GFR can be dissociated from that mediating the increase in RVR, providing functional evidence of angiotensin receptor subtypes in the rat kidney. PMID- 9730778 TI - Antiallergic action of betotastine besilate (TAU-284) in animal models: A comparison with ketotifen. AB - The effects of betotastine besilate (betotastine: TAU-284), a novel antiallergic drug, on homologous passive cutaneous anaphylaxis (PCA), mediator-induced cutaneous reaction, antigen-induced asthmatic responses and platelet-activating factor (PAF)-induced airway eosinophilia in several animal models, were compared to ketotifen. Betotastine (0.1 mg/kg, p.o.) and ketotifen (1 mg/kg, p.o.) inhibited both rat PCA and histamine-induced cutaneous reaction, whereas they showed little effect on serotonin-induced cutaneous reaction. Betotastine (0.3 mg/kg, p.o.) and ketotifen (1 mg/kg, p.o. ) significantly inhibited antigen induced bronchoconstriction in guinea pigs which had been passively sensitized with guinea pig IgE antibody. In actively sensitized guinea pigs, the immediate and late phase increase in airway resistance (Rrs) were observed within 5 min and between 4 and 7 h after the aeroantigen challenge. Betotastine (1 mg/kg, p.o.) inhibited both responses. Ketotifen (1 mg/kg, p.o.) inhibited the immediate phase response, but did not affect the late phase response. Exposure of guinea pigs to aerosolized PAF increased the number of eosinophils in bronchoalveolar lavage fluid 24 h after the stimulation. Betotastine (3-10 mg/kg, p.o.) dose-dependently inhibited PAF-induced accumulation of eosinophils in the bronchoalveolar cavity. In contrast, cetirizine (10 mg/kg, p.o.) showed a tendency to inhibit eosinophil accumulation, and ketotifen (10 mg/kg, p.o.) and terfenadine (10 mg/kg, p.o.) did not have any affect. These results indicate that betotastine could be useful in the treatment of allergic disease such as bronchial asthma. PMID- 9730779 TI - Peripheral benzodiazepine receptor expression on leukocytes and neutrophil function during anticonvulsant monotherapy. AB - Epileptic patients on long-term therapy with a single anticonvulsant showed enhanced expression of peripheral benzodiazepine receptors (pBZrs) on neutrophils, monocytes and lymphocytes. N-Formyl-methionyl-leucyl-phenylalanine induced chemotaxis was significantly impaired in neutrophils from patients on carbamazepine (p < 0.01 vs. controls). Neutrophils from patients on phenytoin had enhanced phorbol myristate acetate-stimulated O-2 production (p < 0. 01 vs. controls) and neutrophils from patients on valproic acid had impaired phagocytosis frequency and Staphylococcus aureus lethality index (p < 0.01 vs. controls). Overexpression of pBZrs on leukocytes may reflect the clinical response to anticonvulsants and may play a role in the immunological effects of some of these drugs. PMID- 9730780 TI - Ca2+-induced inhibition of adenylyl cyclase in turkey erythrocyte membranes. AB - We investigated the effects of calcium ions (Ca2+) on the adenylyl cyclase activity in purified turkey erythrocyte membranes. Results showed the following: (i) Ca2+ inhibits cAMP accumulation stimulated by isoproterenol (1 micromol/l), NaF + AlCl3 (10 mmol/l + 20 micromol/l) or forskolin (10 micromol/l) in EGTA washed turkey erythrocyte membranes. IC50 of free [Ca2+] is approximately 0.1 mmol/l in the presence of Mg2+ (2.5 mmol/l) and isobutylmethylxanthine (1 mmol/l). (ii) The potency of Ca2+ to inhibit cAMP accumulation is independent of the type of stimulus used to activate the adenylyl cyclase. We also evaluated the calcium sensitivity of the basal cAMP accumulation in the presence of GTP (10 micromol/l) and Mg2+ (2.5 mmol/l) which was also inhibited by Ca2+ with the same potency. (iii) The inhibition pattern of cAMP accumulation is not affected by the presence of added calmodulin (100 nmol/l). (iv) Ca2+ is ineffective on the binding of isoproterenol to the beta-adrenoceptors. (v) Increasing the concentration of Ca2+ does not induce an observable activation of cyclic nucleotide phosphodiesterase in the present experimental conditions. Thus, we concluded that the inhibition of cAMP accumulation is due to an inhibition of the adenylyl cyclase rather than the activation of phosphodiesterase(s). The presence of a yet unidentified isoform of adenylyl cyclase that can be directly inhibited by Ca2+ or a Gi protein that can be activated by Ca2+ seems to explain these results. In either case, these results provide an additional mode of cross-talk that can take place between the Ca2+- and cAMP-signaling systems. PMID- 9730781 TI - Whatever happened to delusional perception? AB - Thanks to the analysis of delusional perception's formal binary structure, generations of psychiatrists believed that the problem of delusion, at least from a descriptive point of view, had been resolved. The first-rank Schneiderian symptoms, and delusional perception in particular, had become reference points for the diagnosis of delusion and schizophrenia. Today, however, the phenomenon of delusional perception no longer seems to be taken into serious consideration by psychiatrists and psychopathologists. The crisis of the nosographic specificity of Schneiderian first-rank symptoms and the development of a transnosographic perspective in the study of delusion have definitively loosened the ties that existed in traditional psychopathology between delusional perception, delusion and schizophrenia. Despite the fact that delusional perception has lost its fundamental role in the nosographic sector, it still maintains its importance when studied in an interpersonal context. The significance of formal discontinuity evident in delusional perception authorized the psychiatrist to pause on the threshold of delusion without making allowances for the disastrous lack of comprehension so typical of delusional perception, which in turn must be studied from an interpersonal perspective, not as if it were an aseptic object in a laboratory. The therapeutic relationship can therefore provide an opportunity to recuperate those experiences which the patient was unable to store in his memory and concentrated in delusional perception. PMID- 9730782 TI - Religious faith after psychotic illness. AB - Religion can play an important role in the lives of psychiatric patients. We assessed how often a psychotic illness can lead to a change in the strength of religious faith and how commonly religion is used for coping with such illnesses in a sample of consecutively admitted patients. 52 patients with psychosis were interviewed regarding their religious beliefs after their index admission. 69.4% of the patients were religious, and 11 (22.4%) stated that religion was the most important part of their lives. 30.4% of the sample described that there had been an increase in their religiousness after the onset of illness. 61.2% of patients were using their religion for coping with the illness. Such patients had a better insight into their illness and were more compliant with antipsychotic medication. We conclude that the experience of a psychotic illness is likely to lead to an increase in religious beliefs. Such beliefs are commonly used for coping with the illness and some patients attach a great importance to them. PMID- 9730783 TI - Pathological fear of cot death. AB - Cot death (sudden infant death syndrome) is one of the most common causes of death in the first year of life. Four cases with a pathological fear of cot death are presented. All the patients were depressed and in 2 cases the fear of cot death had an obsessional quality. In all cases there were complications during pregnancy (miscarriage, threatened abortion, recurrent vomiting in last trimester). In 1 case, the patient knew 3 mothers who had suffered cot deaths; in another, the infant was gravely ill in the neonatal period. Pathological fear of cot death can be recognised by the presence of two central features - overvigilance and excessive nocturnal checking of the baby's breathing. Therapeutic interventions are discussed. PMID- 9730784 TI - Parapartum mental illness: a long-term follow-Up study. AB - All mothers (n = 79) in the county of Stockholm who gave birth to a child during 1976-77 and were also hospitalised for the first time in a psychiatric clinic were followed up after a mean interval of 15 years. The sample was classified according to the Research Diagnostic Criteria. Comparisons were made with matched obstetric controls. Five patients had died. The recurrence rate was 51% and 7. 3% relapsed after a subsequent childbirth. No difference was found between psychotic and non-psychotic mothers regarding mean sick-leave days per year. The majority of the women with a depressive disorder at index admission suffered from a minor depressive disorder. The women in the group with an unspecified functional psychosis showed a less severe course of illness than the women in the schizophrenia group. PMID- 9730785 TI - First hospitalized versus rehospitalized patients with an affective disorder or schizophrenia: differences in long-term course and outcome. AB - A comparison of course and outcome in schizophrenics and patients with unipolar affective disorders revealed significant differences not only between the two groups but also between first hospitalized and rehospitalized patients within each group. While schizophrenics fared worse in almost all parameters at the end of the 14-year follow-up period, within each group overall course and outcome were also poorer for rehospitalized versus first hospitalized patients. The poorest course was shown by rehospitalized schizophrenics. Future studies on course and outcome should differentiate between first and rehospitalized patients. PMID- 9730786 TI - Affective disorders, hospital admissions, and seasonal variation of mania in a subtropical area, southern hemisphere. AB - Hospital admissions (n = 15,450) to a state psychiatric hospital in Botucatu, Sao Paulo State, Brazil, over a 10-year period (1982-1991) were reviewed. 157 (1%) patients received a probable diagnosis of affective disorder according to DSM-III R criteria. Among them, 46% had been diagnosed by the staff psychiatrists, and their diagnoses were sustained by the researchers, whereas 54% were diagnosed only by one of the researchers (F.K.C.). These last patients had previously received a diagnosis of paranoid schizophrenia or unspecified psychosis (ICD-9). Most of the patients with affective disorders were bipolar: 72 and 8%, respectively, presented manic and depressive episodes. Thus, only 20% received a diagnosis of major depression. A seasonal pattern in hospital admission was observed only for mania in women, their episodes occurring more often (p < 0.02) in spring and summer. No significant seasonal pattern in hospital admission for depression was found. PMID- 9730787 TI - Delusion of maternity. AB - A few case reports on the delusion of pregnancy can be found in the published literature. However, the delusion of maternity, a delusional conviction of the patient that she is a mother of one or more children, has not yet been studied closely. The author describes the case of a 62-year-old childless patient with a diagnosis of chronic paranoid schizophrenia. She developed a delusion of maternity, persistent for many years. This markedly influenced her behaviour. PMID- 9730788 TI - The modern assessment of personality disorders. Part 1: definition and typology of personality disorders. AB - The general definition of personality disorders (PD) has found agreement in many classification systems and has remained relatively stable over many decades. However, a closer look at the history of the classification of various PD reveals that there are changes from generation to generation: a continuous variation in our cultural norms means that some types are 'renormalised' from time to time and that others are added. On the other hand, a comparison of the descriptions and classifications of PD by Schneider, the DSM and ICD systems shows substantial agreement on many types of PD. PMID- 9730789 TI - The treatment of tuberculosis. PMID- 9730790 TI - Established acute respiratory distress syndrome: benefit of corticosteroid rescue therapy. AB - The mortality of the acute respiratory distress syndrome (ARDS) is still very high. There is some evidence based on case series suggesting that corticosteroids may be beneficial in the chronic fibroproliferative state of the disease. In a retrospective study we analysed the data of 31 non-trauma ARDS patients who had been on mechanical ventilation for at least 7 days. Thirteen cases received corticosteroids at a dosage equivalent to 100-250 mg methylprednisolone at the discretion of the attending physician in charge. Apart from corticosteroid administration, supportive care was identical in the treated and non-treated patient subgroups. Both groups were comparable regarding the relevant demographic, clinical and physiologic data, Apache-II score, radiological score, lung injury score and multiple organ failure score. Mortality in the treatment group was 38% (5/13) as opposed to 67% (12/18) in the untreated group (p = 0.117). There was a significant improvement in the PaO2/FIO2 ratio from a median of -26 to +5 mm Hg measured in a 48-hour period before and after corticosteroid treatment (p = 0.039). The response to corticosteroid therapy could not be predicted on the basis of clinical or physiologic data. Five patients responded within 48 h, and 3 showed a delayed response after 5-6 days. There were no significant complications attributable to corticosteroid treatment. CONCLUSIONS: Although the data of this first comparative study were retrospective, they suggest a beneficial effect of corticosteroid treatment in patients with established fibroproliferative ARDS. A prospective clinical trial, however, is highly warranted in order to fully elucidate the true effect of this therapeutic approach under controlled conditions. PMID- 9730791 TI - Effect of dietary supplementation with polyunsaturated fatty acids on bronchial hyperreactivity in subjects with seasonal asthma. AB - Dietary supplementation with omega-3 essential fatty acids results in the production of uniqe 5-lipoxygenase and cyclooxygenase products which are biologically less active and may inhibit the production, or actions, of the eicosanoids produced when arachidonic acid is the substrate for 5-lipoxygenase and cyclooxygenase, rather than omega-3 essential fatty acids. Since airway inflammation may play a central role in the pathophysiology of asthma, we studied the effect of omega-3 essential fatty acids on bronchial responsiveness in 7 atopic patients suffering from seasonal asthma due to airborne allergens, and positive to intracutaneous skin reaction to two or more allergens. Bronchial responsiveness to ultrasonically nebulized distilled water (UNDW) was determined 30 days from the initial ingestion of 3 g/day of omega-3 essential fatty acids and 30 days after stopping dietary supplementation. Flow volume curves and Raw were recorded before the provocation test, at the end of inhalation, and at 10-, 20-, 30- and 60-min intervals. The maximum fall in forced expiratory volume in 1 s (FEV1) and the maximum increase in airway resistance (Raw) were chosen as the main outcome parameters. After 30 days of dietary supplementation, bronchial responsiveness to UNDW was significantly improved (in fact maximum fall in FEV1 was -11% vs. -28% before treatment, and maximum increase in Raw was +37% vs. +265% before treatment). The challenge test repeated 30 days after stopping dietary supplementation was the same as that recorded before treatment. The present data strongly suggest the hypothesis that dietary supplementation with omega-3 essential fatty acids could decrease bronchial hyperreactivity in atopic patients. PMID- 9730792 TI - Atopy, airway reactivity and compressed air diving in males. AB - A decline in expiratory flow rates in divers has recently been attributed to chronic exposure to hyberbaric air. Airway hyperresponsiveness (AHR) to stimuli due to a hyperbaric environment may play a certain role in this context. The aim of this study was to determine the prevalence of AHR in compressed air divers and to assess the value of bronchial challenges for prediction of fitness to dive. A cross-sectional sample of 59 healthy male volunteers--28 divers and 31 diving candidates (controls)--who had been found fit to dive in a diving medical examination underwent additional allergy screening (skin prick and serum IgE) and a histamine bronchial challenge. Pre- and postchallenge body plethysmography was completed to assess AHR. AHR to histamine was significantly increased among divers and positively related to diving experience whereas divers and controls did not differ significantly with respect to age, anthropometric data, current smoking habits, skin prick reaction, and elevated serum IgE. Our results indicate an increased prevalence of AHR to nonspecific inhalation stimuli in experienced divers. Bronchial challenge tests may be helpful to detect asthmatics in the medical assessment of fitness to dive and for follow-up examinations during a diver's career. PMID- 9730793 TI - A new beclomethasone dipropionate multidose powder inhaler in the treatment of bronchial asthma. AB - The clinical efficacy, tolerability and acceptability of a new multidose powder inhaler (MDPI) containing beclomethasone dipropionate (BDP) were compared with those of a BDP aerosol administered with a large volume spacer (MDI-spacer) among adult asthmatics currently receiving from 500 to 1,000 microgram/day of an inhaled corticosteroid. During the study, the dosage of BDP from both devices was 400 microgram twice daily. Ninety-one patients were randomized to the MDPI group and 42 to the MDI-spacer group. The trial was performed as an open, randomized, parallel group multicenter study. The duration of the treatment period was 12 weeks, and the study was preceded by a 2-week run-in period. During the run-in period, the mean morning peak expiratory flow (PEF) was 487 and 466 1/min in the MDPI and MDI-spacer groups, respectively. After the 12-week treatment, the morning PEF was 491 1/min in the MDPI group and 463 1/min in the MDI-spacer group. The evening values were 500 and 479 1/min during the run-in period and 496 and 476 1/min after the 12-week treatment, respectively. Asthma symptom scores and the use of rescue medication were low in both groups, indicating good efficacy of the preparations tested. The median dose of histamine required to decrease forced expiratory volume in 1 s by 15% increased during the study from 800 to 1,098 microgram in the MDPI group and from 795 to 960 microgram in the MDI spacer group. The most frequent adverse events in both groups were hoarseness and sore throat. There were no statistically significant differences between the treatment groups in serum cortisol values or in the number of patients with thrush. Seventy-two percent of the patients regarded the MDPI easier to use while 95% considered it more portable. Over 80% of the patients felt that the MDPI was also easier to clean and as easy or easier to learn to use than the MDI-spacer. To conclude, the novel powder inhaler is well tolerated and at least equally effective as the conventional MDI-spacer combination in the treatment of asthma with BDP. However, in everyday use, patients clearly favored the powder inhaler. PMID- 9730794 TI - Utility of transfer factor to detect different bronchodilator responses in patients with chronic obstructive pulmonary disease. AB - Previous studies have described that there are different types of disease in patients with established chronic obstructive pulmonary disease (COPD) with different clinical course and functional responses. The aim of this study was to evaluate if the presence of low transfer factor (LTF) values can predict the effectiveness of bronchodilator therapy, and to assess whether this group has different risk factors that may be related with the responses. Eighty patients with COPD were evaluated on three occasions. Initial assessment included a standard respiratory questionnaire, blood analysis, skin prick test and baseline lung function, all performed on the first visit. Bronchodilator response was evaluated after low (0.2 mg) and high (1 mg) doses of salbutamol, and after 2 weeks of oral prednisone. In patients with normal TLCO/VA % (NTF), a higher proportion of subjects with previous history of atopy was the only statistically significant difference compared to those with LTF (odds ratio 4.33; 95% confidence interval 1.06-25.15). Although the mean response in forced expiratory volume in 1 s (FEV1) to treatment was analogous in both groups, when bronchodilation was expressed as percent of predicted, there was a clear trend to a lower response in patients with LTF (0.2 mg salbutamol: 6.99 +/- 5.64 vs. 8.94 +/- 6. 61, p = 0.15; 1 mg salbutamol: 10.18 +/- 6.37 vs. 13.45 +/- 7.90, p < 0.05; oral prednisone: 5.51 +/- 6.94 vs. 8.74 +/- 10.81, p = 0.06). The percentage of patients who had >12% improvement from that predicted in FEV1 was also lower in this group (42 vs. 72%; p < 0. 05). Moreover, TLCO/VA% was significantly lower in those subjects with a negative bronchodilator trial with salbutamol (68 +/- 25 vs. 81 +/- 26; p < 0.05) and prednisone (69 +/- 26 vs. 90 +/- 22; p < 0. 01). In patients with LTF and NTF, airway responsiveness was only significantly related with basal airflow limitation (LTF, r = 0.44; NTF, r = 0.38). All other interaction terms were not statistically significant. These results indicate that in patiens with similar serverity of COPD, the presence of LTF indicates a decreased probability of a positive bronchodilator response, probably reflecting different pathological lesions. We suggest that transfer factor should be taken into consideration when bronchial response is evaluated in large clinical trials. PMID- 9730795 TI - Local and peripheral plasma endothelin-1 in pulmonary hypertension secondary to chronic obstructive pulmonary disease. AB - Endothelin-1 (ET-1) has been described to have crucial effects in the initiation and evolution of pulmonary hypertension (PH) secondary to cardiac disorders. However, the precise role of ET-1 in PH induced by chronic obstructive pulmonary disease (COPD) is not yet clear. The objective of this cross-sectional study was to determine the local and peripheral plasma ET-1 profile of COPD patients with or without PH. Twenty-six COPD patients with clinical and/or laboratory findings suspicious of PH, and 20 healthy smoker volunteers constituted the study population. Patients were allocated to PH (n = 17) and non-PH (n = 9) groups according to their pulmonary artery pressures determined by right-heart catheterization. Plasma ET-1 samples, obtained from the main pulmonary artery (mixed venous blood) and peripheral blood (radial artery and brachial vein), were assessed by radioimmunoassay. Brachial vein ET-1 levels were within normal ranges in PH (2.7 +/- 0.5 pg/ml) and non-PH (3.2 +/- 0. 7 pg/ml) COPD patients compared with that of the controls (4.4 +/- 0. 1 pg/ml). Likewise, radial artery ET-1 levels in PH (3.3 +/- 0.7 pg/ml) and non-PH (2.9 +/- 0.8 pg/ml) groups, and in controls (3.4 +/- 1.1 pg/ml) were also comparable. The pulmonary artery ET-1 concentration of the PH group (13.6 +/- 3.7 pg/ml) was higher than that of the non-PH group (2.2 +/- 0.4 pg/ml) and that of the peripheral blood levels of controls. Elevated pulmonary artery ET-1 in the PH group was inversely correlated only with PaO2 levels. These results could be taken as an evidence of a local role of ET-1 in COPD-induced PH, but it remains to be clarified whether ET-1 is a marker or a mediator of PH in COPD. PMID- 9730796 TI - Elevated serum sialyl Lewis X-i antigen levels in non-small cell lung cancer with lung metastasis. AB - To evaluate the relationship between serum levels of sialyl Lewis X-i antigen and lung metastasis in patients with non-small cell lung cancer (NSCLC), we measured the serum level of the antigen in 299 patients with untreated locally advanced or metastatic NSCLC. Before treatment, serum levels of sialyl Lewis X-i antigen were significantly higher in patients with lung metastasis than in those without lung metastatis (p = 0.0001). Of 201 patients without lung metastasis at the time of primary diagnosis, 121 patients died between July 1987 and December 1995. Serum levels of sialyl Lewis X-i antigen in 21 patients who developed lung metastasis during the period were significantly higher than those in 100 patients who did not develop lung metastasis (p = 0.0171). Our results suggested that sialyl Lewis X-i antigen might be a good indicator for the presence or development of lung metastasis, and it might provide clinical information about the management of patients with NSCLC. PMID- 9730797 TI - Cytokine responses in patients with pneumonia caused by Chlamydia or Mycoplasma. AB - The serum concentrations of interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), and interferon-gamma (IFN-gamma) were measured by enzyme immunoassays in 44 patients with Chlamydia (n = 13) or Mycoplasma (n = 14) pneumonia or influenza A infection (n = 17) and in 20 control subjects. The levels of IFN-gamma were raised in 29/44 patients. The concentrations of IL-6 were raised in 32/44 patients. Raised levels of TNF-alpha were seen in 26/44 but there was no significant difference between the levels of the different groups of patients. All three cytokines indicated clinical recovery when acute and convalescent samples from 10 patients with Chlamydia pneumonia were analyzed. IFN gamma, IL6 and TNF-alpha are present in the circulation in the majority of patients with Chlamydia and Mycoplasma pneumonia and in influenza A infection. We suggest that repeated measurement of cytokines, such as IL-6, IFN-gamma and TNF alpha, may be useful in the management of lower respiratory tract infections but further studies are needed to define the value of cytokine measurements in acute pneumonia. PMID- 9730798 TI - Effectiveness of ethanolamine oleate as a pleural sclerosing agent in rabbits. AB - The ideal pleural sclerosing agent should be easily administered, without significant side effects, inexpensive, and widely available. None of the agents presently used meets all of these criteria. Ethanolamine oleate (ETH) is a sclerosing agent used in the sclerotherapy treatment of varicose veins of the legs and esophagus. The objective of the present study was to assess the efficacy of ETH as a pleural sclerosant in rabbits. An additional objective was to assess if better results were obtained when dextrose 50% (D50) as opposed to saline was used as the diluent. Each group of 10 rabbits received a total volume of 2 ml intrapleurally. The eight treatments were as follows: (1) 2 ml saline; (2) 2 ml D50; (3) 25 mg ETH plus 1.5 ml saline; (4) 25 mg ETH plus 1.5 ml D50; (5) 50 mg ETH plus 1.0 ml saline; (6) 50 mg ETH plus 1 ml D50; (7) 75 mg ETH plus 0.5 ml D50, and (8) 100 mg ETH. The rabbits were sacrificed 28 days after the injection. The intrapleural instillation of ETH resulted in evident pleurodesis, which was dose-dependent; 100 mg ETH induced significantly (p<0.05) more adhesions than did any other treatment. The selection of the diluent had no effect on the pleurodesis. The microscopic examination of the right visceral pleura showed that the mean degree of fibrosis after 100 mg ETH was significantly (p<0.05) greater than that after the other solutions. The mean degree of pleural inflammation, lung inflammation and lung fibrosis was minimal in all the groups. From this study we conclude that undiluted ETH produces pleurodesis in our experimental model. At the doses used, the pleurodesis was less than that produced after talc, tetracycline derivatives or silver nitrate in the same model. PMID- 9730799 TI - Single pulmonary nodule after cardiac catheterization. PMID- 9730800 TI - A syndrome of inappropriate secretion of antidiuretic hormone associated with pleuritis caused by OK-432. AB - We here report a case presenting with the syndrome of inappropriate secretion of antidiuretic hormone (SIADH) after having been treated for pleurodesis with OK 432, which is a lyophilized preparation of an attenuated strain of Streptococcus pyogenes. The patient, who had undergone a subtotal esophagectomy 4 years previously, was referred to our department after the diagnosis of a metastatic lung tumor. A right lower lobectomy of the lung was performed, and prolonged air leakage from a pulmonary fistula thereafter developed because of the dissection of severe pleural adhesion. OK-432 (5 klinische einheiten) was administered to the pleural cavity 3 times. On the 13th postoperative day, the patient began to complain of general fatigue and nausea. SIADH was diagnosed based on laboratory findings such as hyponatremia, serum hypo-osmolality and a high excretion of sodium in the urine. A restriction of the fluid intake with a sodium supplement resulted in the return to a normal serum level within 2 weeks. We therefore concluded that the intrapleural instillation of OK-432 had apparently caused SIADH in this case, because no other causes could be found. PMID- 9730801 TI - Eight-year follow-up study of a patient with central alveolar hypoventilation treated with diaphragm pacing. AB - We had treated a 17-year-old girl with central alveolar hypoventilation with diaphragm pacing at home for the past 8 years. Nocturnal diaphragm pacing with an open tracheostomy was effective in maintaining an adequate ventilation after 4 months of undergoing training of the diaphragm on the pacemaker implantation. However, the diaphragm pacemaker failed to maintain it mostly in the presence of respiratory tract infection, when she was treated with intermittent positive pressure ventilation. Pulse oximetry was used at home for monitoring the adequacy of respiratory support. We conclude that the respiratory assistance by the diaphragm pacemaker or the use of a mechanical ventilator as a backup was highly useful for the home care of a patient with central alveolar hypoventilation. PMID- 9730802 TI - Decompensated cor pulmonaleas the first manifestation of adult-onset myopathy. AB - A 46-year-old white female was admitted for decompensated cor pulmonale (CP). It had not interfered with her daily activities and she had not experienced shortness of breath, fatigue or muscle weakness prior to the onset of right heart failure. A thorough investigation revealed severe generalized muscle weakness with restrictive chest bellows disease and secondary CP (mean pressure in the pulmonary artery 60 mm Hg). After having refused respiratory support the patient died a few days after admission. The muscle biopsy was consistent with adult onset acid-maltase deficiency. This is a rare case of metabolic myopathy presenting as decompensated CP without previous symptoms of muscle weakness. This condition can easily be treated with nocturnal ventilatory support, improving the quality and length of life. PMID- 9730803 TI - Adverse reactions to peak flow monitoring. Report of 5 patients. AB - Five patients with adverse reactions to peak flow monitoring are presented: 2 patients had herniation of abdominal content, while the others presented with vasovagal syncope, minor depression and neurotic preoccupation with peak flow values, respectively. As a result, 3 of the 5 patients became noncompliant. For nonpsychological somatic adverse reactions, we calculated an incidence of 1.1 cases/1,000 patients started on peak flow monitoring. Adverse reactions with a psychological background may be more frequent. Clinicians should bear in mind that patients noncompliant with peak flow monitoring may have discontinued because of adverse reactions. PMID- 9730804 TI - Chronic eosinophilic pneumonia with atypical radiographic presentation. AB - Chronic eosinophilic pneumonia (CEP) is a rare disease characterized by a progressive symptomatic deterioration of more than 1 month, pulmonary infiltrates with eosinophils, and a dramatic response to corticosteroid treatment. We describe a patient with CEP who presented with right upper lung infiltrates with evidence of cavitation. PMID- 9730805 TI - Rhodococcus equi lung abscess complicating Evan's syndrome treated with corticosteroid. AB - We report a case of Rhodococcus equi, an unusual pathogen, causing a right upper lobe lung abscess in a patient with Evan's syndrome (auto-immune haemolytic anaemia and thrombocytopenia) who was treated with high-dose corticosteroid therapy. The patient was treated successfully with clarithromycin, vancomycin, ciprofloxacin and imipenen which appear to be effective in combination for this unusual condition in which the treatment regimen has been controversial. PMID- 9730806 TI - Hypercalcemia and multiple pulmonary nodules. PMID- 9730808 TI - NMR-Based model of a telomerase-inhibiting compound bound to G-quadruplex DNA. AB - The single-stranded (TTAGGG)n tail of human telomeric DNA is known to form stable G-quadruplex structures. Optimal telomerase activity requires the nonfolded single-stranded form of the primer, and stabilization of the G-quadruplex form is known to interfere with telomerase binding. We have identified 3,4,9, 10 perylenetetracarboxylic diimide-based ligands as potent inhibitors of human telomerase by using a primer extension assay that does not use PCR-based amplification of the telomerase primer extension products. A set of NMR titrations of the ligand into solutions of G-quadruplexes using various oligonucleotides related to human telomeric DNA showed strong and specific binding of the ligand to the G-quadruplex. The exchange rate between bound and free DNA forms is slow on the NMR time scale and allows the unequivocal determination of the binding site and mode of binding. In the case of the 5' TTAGGG sequence, the ligand-DNA complex consists of two quadruplexes oriented in a tail-to-tail manner with the ligand sandwiched between terminal G4 planes. Longer telomeric sequences, such as TTAGGGTT, TTAGGGTTA, and TAGGGTTA, form 1:1 ligand-quadruplex complexes with the ligand bound at the GT step by a threading intercalation mode. On the basis of 2D NOESY data, a model of the latter complex has been derived that is consistent with the available experimental data. The determination of the solution structure of this telomerase inhibitor bound to telomeric quadruplex DNA should help in the design of new anticancer agents with a unique and novel mechanism of action. PMID- 9730809 TI - Active serine involved in the stabilization of the active site loop in the Humicola lanuginosa lipase. AB - We have investigated the binding properties of and dynamics in Humicola lanuginosa lipase (Hll) and the inactive mutant S146A (active Ser146 substituted with Ala) using fluorescence spectroscopy and molecular dynamics simulations, respectively. Hll and S146A show significantly different binding behavior for phosphatidylcholine (PC) and phosphatidylglycerol (PG) liposomes. Generally, higher binding affinity is observed for Hll than the S146A mutant. Furthermore, depending on the matrix, the addition of the transition state analogue benzene boronic acid increases the binding affinity of S146A, whereas only small changes are observed for Hll suggesting that the active site lid in the latter opens more easily and hence more lipase molecules are bound to the liposomes. These observations are in agreement with molecular dynamics simulations and subsequent essential dynamics analyses. The results reveal that the hinges of the active site lid are more flexible in the wild-type Hll than in S146A. In contrast, larger fluctuations are observed in the middle region of the active site loop in S146A than in Hll. These findings reveal that the single mutation (S146A) of the active site serine leads to substantial conformational alterations in the H. lanuginosa lipase and different binding affinities. PMID- 9730810 TI - MutY DNA glycosylase: base release and intermediate complex formation. AB - MutY protein, a DNA glycosylase found in Escherichia coli, recognizes dA:dG, dA:8 oxodG, and dA:dC mismatches in duplex DNA, excising the adenine moiety. We have investigated the mechanism of action of MutY, addressing several points of disagreement raised by previous studies of this enzyme. MutY forms a covalent intermediate with its DNA substrate but does not catalyze strand cleavage. The covalent intermediate has a half-life of approximately 2.6 h, 2 orders of magnitude greater than the half-life of Schiff bases formed when E. coli formamidopyrimidine-DNA glycosylase (Fpg) and endonuclease III react with their respective substrates. The covalent complex between MutY and its DNA substrate involves Lys-142; however, the position of this residue in the presumptive active site differs from that of catalytic residues involved in Schiff base formation associated with endonuclease III and related DNA glycosylases/AP lyases. MutY converts DNA duplexes containing the dA:8-oxodG mispair to a product containing an abasic site; heat-induced cleavage of this product may account for the several reports in the literature that ascribe AP lyase activity to MutY. The MutY-DNA intermediate complex is highly stable and hinders access by Fpg to DNA, thereby avoiding a double-strand break. Cross-linking of MutY to DNA may play an important role in the regulation of base excision repair. PMID- 9730811 TI - Mechanism of phospholipid binding by the C2A-domain of synaptotagmin I. AB - Synaptotagmin I is a synaptic vesicle membrane protein that probably functions as a Ca2+ sensor in neurotransmitter release and contains two C2-domains which bind Ca2+. The first C2-domain of synaptotagmin I (the C2A-domain) binds phospholipids in a Ca2+-dependent manner similar to that of the C2-domains of protein kinase C, cytoplasmic phospholipase A2, and phospholipase Cdelta1. Although the tertiary structure of these C2-domains is known, the molecular basis for their Ca2+ dependent interactions with phospholipids is unclear. We have now investigated the mechanisms involved in Ca2+-dependent phospholipid binding by the C2A-domain of synaptotagmin I. Our data show that the C2A-domain binds negatively charged liposomes in an electrostatic interaction that is determined by the charge density of the liposome surface but not by the phospholipid headgroup. At the tip of the C2A-domain, three tightly clustered Ca2+-binding sites are formed by five aspartates and one serine. Mutations in these aspartate and serine residues demonstrated that all three Ca2+-binding sites are required for phospholipid binding. The Ca2+ binding sites at the top of the C2A-domain are surrounded by positively charged amino acids that were shown by mutagenesis to be also involved in phospholipid binding. Our results yield a molecular picture of the interactions between a C2-domain and phospholipids. Binding is highly electrostatic and occurs between the surfaces of the phospholipid bilayer and of the tip of the C2A-domain. The data suggest that the negatively charged phospholipid headgroups interact with the basic side chains surrounding the Ca2+ binding sites and with bound Ca2+ ions, thereby filling empty coordination sites and increasing the apparent affinity for Ca2+. In addition, insertion of hydrophobic side chains may contribute to phospholipid binding. This model is likely to be general for other C2-domains, with the relative contributions of electrostatic and hydrophobic interactions dictated by the exposed side chains surrounding the Ca2+-binding region. PMID- 9730812 TI - Crystal structure of the zinc-dependent beta-lactamase from Bacillus cereus at 1.9 A resolution: binuclear active site with features of a mononuclear enzyme. AB - The structure of the zinc-dependent beta-lactamase II from Bacillus cereus has been determined at 1.9 A resolution in a crystal form with two molecules in the asymmetric unit and 400 waters (space group P3121; Rcryst = 20.8%). The active site contains two zinc ions: Zn1 is tightly coordinated by His86, His88, and His149, while Zn2 is loosely coordinated by Asp90, Cys168, and His210. A water molecule (W1) lies between the two zinc ions but is significantly closer to Zn1 and at a distance of only 1.9 A is effectively a hydroxide moiety and a potential, preactivated nucleophile. In fact, Asp90 bridges W1 to Zn2, and its location is thus distinct from that of the bridging water molecules in the binuclear zinc peptidases or other binuclear zinc hydrolases. Modeling of penicillin, cephalosporin, and carbapenem binding shows that all are readily accommodated within the shallow active site cleft of the enzyme, and the Zn1 bound hydroxide is ideally located for nucleophilic attack at the beta-lactam carbonyl. This enzyme also functions with only one zinc ion present. The Zn1-Zn2 distances differ in the two independent molecules in the crystal (3.9 and 4.4 A), yet the Zn1-W1 distances are both 1.9 A, arguing against involvement of Zn2 in W1 activation. The role of Zn2 is unclear, but the B. cereus enzyme may be an evolutionary intermediate between the mono- and bizinc metallo-beta-lactamases. The broad specificity of this enzyme, together with the increasing prevalence of zinc-dependent metallo-beta-lactamases, poses a real clinical threat, and this structure provides a basis for understanding its mechanism and designing inhibitors. PMID- 9730813 TI - Solution structure of two new toxins from the venom of the Chinese scorpion Buthus martensi Karsch blockers of potassium channels. AB - The solution structure of BmTX2 purified from the venom of the Chinese Buthid Buthus martensi has been determined by 2D NMR spectroscopy techniques which led to the description of its 3D conformation. The structure consists of a triple stranded beta-sheet connected to a helical structure. This helix encompasses 10 residues, from 11 to 20, begins with a turn of 310 helix, and ends with an alpha helix. The three strands of beta sheet comprise residues 2-6, with a bulge covering residues 4 and 5, 26-29, and 32-35, with a type I' beta turn centered on residues 30-31. We also characterized the solution structure of BmTX1. The two toxins which are potent blockers of both large-conductance calcium-activated potassium channels (BKCa channels) and voltage-gated potassium channels (Kv1. 3) are highly superimposable and possess the same structural characteristics. Analysis of these structures allows us to hypothesize that, besides the main surface of interaction described by the functional map of charybdotoxin, one can expect that the binding of scorpion toxins on BKCa channels may involve residues on the edge of this surface. PMID- 9730814 TI - Structure and interaction site of the regulatory domain of troponin-C when complexed with the 96-148 region of troponin-I. AB - The structure of the regulatory domain of chicken skeletal troponin-C (residues 1 90) when complexed with the major inhibitory region (residues 96-148) of chicken skeletal troponin-I was determined using multinuclear, multidimensional NMR spectroscopy. This complex represents the first interaction formed between the regulatory domain of troponin-C and troponin-I after calcium binding in the regulation of muscle contraction. The stoichiometry of the complex was determined to be 1:1, with a dissociation constant in the 1-40 microM range. The structure of troponin-C in the complex was calculated from 1039 NMR distance and 111 dihedral angle restraints. When compared to the structure of this domain in the calcium saturated "open" form but in the absence of troponin-I, the bound structure appears to be slightly more "closed". The troponin-I peptide-binding site was found to be in the hydrophobic pocket of calcium saturated troponin-C, using edited/filtered NMR experiments and chemical shift mapping of changes induced in the regulatory domain upon peptide binding. The troponin-I peptide (residues 96-148) was found to bind to the regulatory domain of troponin-C very similarly, but not identically, to a shorter troponin-I peptide (region 115-131) thought to represent the major interaction site of troponin-I for this domain of troponin-C. PMID- 9730815 TI - The structural origin of nonplanar heme distortions in tetraheme ferricytochromes c3. AB - Resonance Raman (RR) spectroscopy, molecular mechanics (MM) calculations, and normal-coordinate structural decomposition (NSD) have been used to investigate the conformational differences in the hemes in ferricytochromes c3. NSD analyses of heme structures obtained from X-ray crystallography and MM calculations of heme-peptide fragments of the cytochromes c3 indicate that the nonplanarity of the hemes is largely controlled by a fingerprint peptide segment consisting of two heme-linked cysteines, the amino acids between the cysteines, and the proximal histidine ligand. Additional interactions between the heme and the distal histidine ligand and between the heme propionates and the protein also influence the heme conformation, but to a lesser extent than the fingerprint peptide segment. In addition, factors that influence the folding pattern of the fingerprint peptide segment may have an effect on the heme conformation. Large heme structural differences between the baculatum cytochromes c3 and the other proteins are uncovered by the NSD procedure [Jentzen, W., Ma, J.-G., and Shelnutt, J. A. (1998) Biophys. J. 74, 753-763]. These heme differences are mainly associated with the deletion of two residues in the covalently linked segment of hemes 4 for the baculatum proteins. Furthermore, some of these structural differences are reflected in the RR spectra. For example, the frequencies of the structure-sensitive lines (nu4, nu3, and nu2) in the high frequency region of the RR spectra are lower for the Desulfomicrobium baculatum cytochromes c3 (Norway 4 and 9974) than for the Desulfovibrio (D.) gigas, D. vulgaris, and D. desulfuricans strains, consistent with a more ruffled heme. Spectral decompositions of the nu3 and nu10 lines allow the assignment of the sublines to individual hemes and show that ruffling, not saddling, is the dominant factor influencing the frequencies of the structure-sensitive Raman lines. The distinctive spectra of the baculatum strains investigated are a consequence of hemes 2 and 4 being more ruffled than is typical of the other proteins. PMID- 9730816 TI - Protein denaturation: a small-angle X-ray scattering study of the ensemble of unfolded states of cytochrome c. AB - Solution X-ray scattering was used to study the equilibrium unfolding of cytochrome c as a function of guanidine hydrochloride concentration at neutral pH. The radius of gyration (Rg) shows a cooperative transition with increasing denaturant with a similar Cm to that observed with circular dichroism. However, the lack of an isoscattering point in the X-ray scattering patterns suggests the equilibrium unfolding is not simply a two-state process. Singular value decomposition (SVD) analysis was applied to the scattering patterns to determine the number of distinct scattering species. SVD analysis reveals the existence of three components, suggesting that at least three equilibrium states of the protein exist. A model was employed to determine the thermodynamic parameters and the scattering profiles of the three equilibrium states. These scattering profiles show that one state is native (N). The other two states (U1, U2) are unfolded, with U2 being fully unfolded and U1 having some residual structure. Using the thermodynamic parameters to calculate fractional populations, U1 is maximally populated at intermediate denaturant concentrations while U2 is maximally populated at high denaturant concentrations. It is likely that there is a multiplicity of denatured states with U1 and U2 representing an average of the denatured states populated at intermediate and high denaturant concentrations, respectively. PMID- 9730818 TI - Structural changes in the heme proximal pocket induced by nitric oxide binding to soluble guanylate cyclase. AB - When expressed in Escherichia coli, the heme domain [beta1(1-385)] of rat lung soluble guanylate cyclase (sGC) is isolated with a stoichiometric amount of bound heme [Zhao, Y., and Marletta, M. A. (1997) Biochemistry 36, 15959-15964]. Nitric oxide (NO) binding to the heme in beta1(1-385) leads to cleavage of the Fe-His bond and formation of a five-coordinate NO-heme complex. Addition of imidazole to the five-coordinate NO complex shifts the Soret peak from 399 to 420 nm, which appears to result from the formation of a six-coordinate NO complex. Removal of the added imidazole by gel filtration results in formation of the five-coordinate NO complex once again. The EPR spectrum of the putative six-coordinate NO complex has nine distinct derivative-shaped lines (a triplet of triplets), which is the signature spectrum of a six-coordinate NO complex with two nitrogen atoms as the axial ligands. [15N]Imidazole simplifies the six-coordinate NO complex EPR spectrum to six distinct derivative-shaped lines (a triplet of doublets), indicating that the other axial ligand in the six-coordinate NO complex is an imidazole molecule. These results show that NO binding to sGC not only leads to the cleavage of the Fe-His bond but also induces a conformational change which opens the heme proximal pocket large enough to accommodate an exogenous imidazole molecule. These observations have important implications for determining the NO activation mechanism of sGC. PMID- 9730817 TI - Imidazole is a sensitive probe of steric hindrance in the distal pockets of oxygen-binding heme proteins. AB - The FixL heme-based sensor, despite its low affinity for oxygen, is much more reactive than myoglobin toward the large polar ligand imidazole. To determine which features of a myoglobin heme pocket favor binding of imidazole, we have measured binding of this ligand to the FixL heme domain, elephant myoglobin, wild type sperm whale myoglobin, and sperm whale myoglobins having alanine, valine, threonine, glutamine, leucine, phenylalanine, or tryptophan substitutions of the distal (E7) histidine residue. Except for histidine, the association rate constants dropped more than 3000-fold as the volume of the E7 side chain, at position 64, was expanded from alanine (10(6) M-1 s-1) to phenylalanine (10(3) M 1 s-1). There was inhibition of imidazole binding due to displacement of coordinated water from H64 and H64Q sperm whale myoglobins, where the E7 side chain hydrogen bonds directly to the bound ligand. The imidazole dissociation rate constants varied less dramatically and less consistently with any single factor, though they were measurably decreased by hydrogen bonding to an E7 glutamine or histidine. On the whole, the results for the sperm whale myoglobin E7 substitutions show that the rate constants for imidazole binding are useful and sensitive indicators of steric hindrance and polar interactions in the distal pockets of myoglobins. The combined effects of the glutamine 64 and phenylalanine 29 in elephant myoglobin largely account for its increased imidazole association and dissociation rate constants, respectively, compared to those of sperm whale myoglobin. An unhindered distal pocket not competent to stabilize positive poles is indicated by the large imidazole association (>/=10(4) M-1 s-1) and dissociation (>/=50 s-1) rate constants, parameters that are characteristic of FixL. PMID- 9730819 TI - Synthesis, biological activity, and conformational analysis of peptidomimetic analogues of the Saccharomyces cerevisiae alpha-factor tridecapeptide. AB - Biochemical and biophysical investigations on the Saccharomyces cerevisiae alpha factor indicate that this tridecapeptide mating pheromone (WHWLQLKPGQPMY) might adopt a type II beta-turn in the center of the peptide when it binds to its G protein-coupled receptor. To test this hypothesis we synthesized analogues of alpha-factor incorporating a (R or S)-gamma-lactam conformational constraint [3 (R or S)-amino-2-oxo-1-pyrrolidineacetamido] in place of the Pro-Gly at residues 8 and 9 of the peptide and tested their biological activities and receptor binding. Analogues were purified to >99% homogeneity as evidenced by high performance liquid chromatography and capillary electrophoresis and characterized by amino acid analysis, mass spectrometry, and nuclear magnetic resonance (NMR) spectroscopy. The restricted alpha-factor analogue WHWLQLK[(R)-gamma lactam]QP[Nle]Y was more active than its lactam-containing diastereomeric homologue WHWLQLK[(S)-gamma-lactam]QP[Nle]Y and about equally active with the [Nle12]-alpha-factor in growth arrest and FUS1-lacZ gene induction assays. Both lactam analogues competed with tritiated [Nle12]-alpha-factor for binding to the alpha-factor receptor (Ste2p) with the (R)-gamma-lactam-containing peptide having 7-fold higher affinity than the (S)-gamma-lactam-containing homologue. Two dimensional NMR spectroscopy and modeling analysis gave evidence that the (R) gamma-analogue is a flexible peptide that assumes a transient gamma-turn structure around the lactam moiety. The results represent the first example of an alpha-factor analogue containing a peptidomimetic constraint that is as active as the native pheromone. The correlation between activity and structure provides further evidence that the biologically active conformation of the molecule contains a turn in the middle of the pheromone. This study provides new insights into the structural basis of alpha-factor activity and adds to the repertoire of conformationally biasing constraints that can be used to maintain and even enhance biological activity in peptide hormones. PMID- 9730820 TI - Self-association of linker histone H5 and of its globular domain: evidence for specific self-contacts. AB - The ability of avian-specific linker histone H5, and the globular domains of H5 (GH5) and H1(0) (GH1(0), to self-associate either free in solution or when bound to DNA was investigated. All three proteins underwent a salt-dependent increase in turbidity that may be indicative of nonspecific interactions. Dithiobis(succinimidyl propionate) cross-linking was used to measure specific contacts for both H5 and GH5 free in solution and bound to DNA. H5 and GH5 each became cross-linked in solution, with GH5 displaying divalent polymerization interactions, which suggests that two specific surfaces were involved in the assembly process. For GH5-DNA complexes, cross-linking appeared to be largely the consequence of aggregation, but under low concentrations of DSP, cross-linking GH5 was observed to assemble preferentially onto DNA before oligomerizing to form massive aggregates. Both linear and supercoiled DNA facilitated GH5 interactions compared to assembly in solution; differences in the distribution of cross-linked polymer sizes indicates that assembly is dependent on both the presence of DNA and the morphology of the DNA. Finally, on the basis of a technique referred to as quantitative proteolysis, GH5 assembly on DNA appears to involve specific protein-protein contacts involving the C terminus of one partner. Overall, the cumulative results reported here support the premise that linker histones assemble specifically both in solution and on DNA. PMID- 9730821 TI - Investigation of the role of tyrosine-114 in the activity of human O6 alkylguanine-DNA alkyltranferase. AB - Tyrosine-114 is one of 13 totally conserved amino acids in all known sequences of O6-alkylguanine-DNA alkyltransferase (AGT). The importance of this amino acid in repair of alkylated DNA by AGT was studied by changing it to phenylalanine (F), alanine (A), threonine (T), or glutamic acid (E) in human AGT. The activities of the mutant proteins were then compared to those of the wild type with regard to abilities to do the following: (a) protect Escherichia coli from the methylating agent N-methyl-N'-nitro-N-nitrosoguanidine (MNNG); (b) repair methylated DNA in vitro; (c) bind to oligodeoxynucleotides containing O6-methylguanine; and (d) react with the low molecular weight pseudosubstrate, O6-benzylguanine. When expressed at high levels in E. coli strain GWR109, lacking endogenous AGT, the wild type and the Y114F mutant were highly effective in reducing mutations and cell killing by MNNG. The Y114A mutant had a much smaller protective effect, and mutants Y114T and Y114E were inactive. Purified preparations of all four AGT mutants showed an approximately similar degree (74-120-fold) of reduction in the rate of reaction with O6-benzylguanine. In contrast, the degree of reduction in activity toward methylated DNA substrates in vitro varied according to the mutation with the more conservative Y114F producing only a 30-fold reduction and the most drastic change of Y114E abolishing activity completely. Alteration Y114A produced a 1000-fold reduction whereas Y114T reduced activity by 10000-fold. All of the mutations affected the binding of AGT to single- or double-stranded oligodeoxynucleotides containing O6-methylguanine. The extent of increase in the Kd varied according to the amino acid with 2-5-fold (F), 7-11-fold (A), 167-200 fold (T), and 600-1000-fold (E) increases. These results are consistent with tyrosine-114 playing a role both in the binding of AGT to its DNA substrate and in facilitating the transfer of the alkyl group. It is probable that AGT resembles other DNA repair proteins in bringing about a "flipping out" of the target base from the DNA helix. Tyrosine-114 is therefore an excellent candidate for a key role in the interaction with the flipped O6-methylguanine. The results also show that when large amounts of AGT are produced in the cell, substantial decreases in the efficiency with which AGT can repair methylated DNA do not prevent the ability to protect E. coli from toxic alkylating agents. Mutant Y114F, whose activity was reduced by 30-fold, was equal to wild-type AGT in bringing about this protection. PMID- 9730822 TI - The 32- and 14-kilodalton subunits of replication protein A are responsible for species-specific interactions with single-stranded DNA. AB - Replication protein A (RPA) is a multisubunit single-stranded DNA-binding (ssDNA) protein that is required for cellular DNA metabolism. RPA homologues have been identified in all eukaryotes examined. All homologues are heterotrimeric complexes with subunits of approximately 70, approximately 32, and approximately 14 kDa. While RPA homologues are evolutionarily conserved, they are not functionally equivalent. To gain a better understanding of the functional differences between RPA homologues, we analyzed the DNA-binding parameters of RPA from human cells and the budding yeast Saccharomyces cerevisiae (hRPA and scRPA, respectively). Both yeast and human RPA bind ssDNA with high affinity and low cooperativity. However, scRPA has a larger occluded binding site (45 nucleotides versus 34 nucleotides) and a higher affinity for oligothymidine than hRPA. Mutant forms of hRPA and scRPA containing the high-affinity DNA-binding domain from the 70-kDa subunit had nearly identical DNA binding properties. In contrast, subcomplexes of the 32- and 14-kDa subunits from both yeast and human RPA had weak ssDNA binding activity. However, the binding constants for the yeast and human subcomplexes were 3 and greater than 6 orders of magnitude lower than those for the RPA heterotrimer, respectively. We conclude that differences in the activity of the 32- and 14-kDa subunits of RPA are responsible for variations in the ssDNA-binding properties of scRPA and hRPA. These data also indicate that hRPA and scRPA have different modes of binding to ssDNA, which may contribute to the functional disparities between the two proteins. PMID- 9730823 TI - The impact of an exocyclic cytosine adduct on DNA duplex properties: significant thermodynamic consequences despite modest lesion-induced structural alterations. AB - The exocyclic base adduct 3,N4-deoxyethenocytosine (epsilonC) is a common DNA lesion that can arise from carcinogen exposure and/or as a biproduct of cellular processes. We have examined the thermal and thermodynamic impact of this lesion on DNA duplex properties, as well as the structural alterations imparted by the lesion. For these studies, we used calorimetric and spectroscopic techniques to investigate a family of 13-mer DNA duplexes of the form (5'CGCATGNGTACGC3')x(3'GCGTACNCATGCG5'), where the central NxN base pair represents the four standard Watson-Crick base pairs (corresponding to four control duplexes), and where either one of the N bases has been replaced by epsilonC, yielding eight test duplexes. Studies on these 12 duplexes permit us to assess the impact of the epsilonC lesion as a function of sequence context. Our spectroscopic and calorimetric data allow us to reach the following conclusions: (i) The epsilonC lesion imparts a large penalty on duplex stability, with sequence context only modestly modulating the extent of this lesion-induced destabilization. This result contrasts with our recent studies of duplexes with abasic sites, where sequence context was found to be the predominant determinant of thermodynamic damage. (ii) For the epsilonC-containing duplexes, sequence context effects are most often observed in the enthalpic contribution to lesion induced duplex destabilization. However, due to compensating entropies, the free energy changes associated with this lesion-induced duplex destablization are nearly independent of sequence context. (iii) Despite significant lesion-induced changes in duplex energetics, our spectroscopic probes detect only modest lesion induced changes in duplex structure. In fact, the overall duplex maintains a global B-form conformation, in agreement with NMR structural data. We discuss possible interpretations of the apparent disparity between the severe thermodynamic and relatively mild structural impacts of the epsilonC lesion on duplex properties. We also note and discuss the implications of empirical correlations between biophysical and biological properties of lesion-containing duplexes. PMID- 9730824 TI - Probing the mechanism of proton coupled electron transfer to dioxygen: the oxidative half-reaction of bovine serum amine oxidase. AB - Bovine serum amine oxidase (BSAO) catalyzes the oxidative deamination of primary amines, concomitant with the reduction of molecular oxygen to hydrogen peroxide via a ping-pong mechanism. A protocol has been developed for an analysis of chemical and kinetic mechanisms in the conversion of dioxygen to hydrogen peroxide. Steady-state kinetics show that two groups need to be deprotonated to facilitate the oxidative half-reaction. The pH dependence of Vmax/Km(O2) reveals pKa's of 6.2 +/- 0.3 and 7.0 +/- 0.2, respectively. A pKa of 7.2 +/- 0.1 has been obtained from a titration of anaerobically reduced BSAO using UV-vis spectrophotometry. The near identity of the pKa obtained from the reduced enzyme titration with the second pKa from steady-state kinetics suggests that this second pKa arises from the reduced cofactor. The assignment of pKa is supported by the observed pH dependence for formation of the cofactor semiquinone signal, detected by EPR spectroscopy under anaerobic conditions. To address the nature of rate-limiting steps in the oxidative half-reaction, the solvent isotope effect, viscosity effect, and O-18 isotope effect on Vmax/Km(O2) have been determined. The solvent isotope effect is indistinguishable from unity, ruling out a proton transfer as a rate-determining step. Use of glucose as a solvent viscosogen shows no viscosity effect, indicating that binding of oxygen is not in the rate determining step. The O-18 kinetic isotope effect is independent of pH with an average value of 18(V/K) = 1.0097 +/- 0. 0010. This has been compared to calculated equilibrium O-18 isotope effects for various dioxygen intermediate species [Tian and Klinman (1993) J. Am. Chem. Soc. 115, 8891], leading to the conclusion that either the first electron transfer to dioxygen or the desorption of product peroxide from a Cu(II)-OOH complex could be the rate-limiting step. The distribution of steady-state enzyme species was, therefore, analyzed through a combination of stopped-flow experiments and analysis of DV and D(V/K) for benzylamine oxidation. We conclude that the major species accumulating in the steady state are the oxidized cofactor-substrate Schiff base complex and the reduced, aminoquinol form of cofactor. These data rule out a slow release of product hydroperoxide from the aminoquinone form of enzyme, leading to the conclusion that the first electron transfer from substrate-reduced cofactor to dioxygen is the rate-determining step in the oxidative half-reaction. This step is also estimated to be 40% rate-limiting in kcat. An important mechanistic conclusion from this study is that dioxygen binding is a separate step from the rate-limiting electron-transfer step to form superoxide. On the basis of a recently determined X-ray structure for the active form of a yeast amine oxidase from Hansenula polymorpha [Li et al. (1998) Structure 6, 293], a hydrophobic space has been identified near the O-2 position of reduced cofactor as the putative dioxygen binding site. Movement of superoxide from this site onto the Cu(II) at the active site may occur prior to further electron transfer from cofactor to superoxide. PMID- 9730825 TI - Surface topography of phytochrome A deduced from specific chemical modification with iodoacetamide. AB - Phytochromes are a photoreversible photochromic light switch for photomorphogenesis in plants. The molecular structure and functional mechanism of phytochromes are not fully understood. On the basis of complete mapping of total tryptic digest of the iodoacetamide-modified oat phytochrome A (phyA), the molecular surface topography of phyA was probed by specific chemical modification of cysteine residues with [14C]iodoacetamide. Under native conditions, only two cysteines (Cys-158 and Cys-311) of eleven half-cystines of the N-terminal chromophore binding domain were modified to a significant extent. In the C terminal domain, six cysteine residues (Cys-715, Cys-774, Cys-809, Cys-869, Cys 961, Cys-995) were readily accessible to iodoacetamide. Among the reactive cysteine residues, only cysteine-311 displayed reactivity that was dependent on the photochromic form (Pr left arrow over right arrow Pfr) of the photoreceptor. Surprisingly, the modification of Cys-311 in the vicinity of the chromophore attachment site (Cys-321) did not have any detectable effect on spectral properties of phyA. Most of the cysteines of the N-terminal domain (Cys-83, Cys 175, Cys-291, Cys-370, Cys-386, Cys-445, Cys-506) are deeply buried in the core of the chromophore binding domain, as they can be modified only after denaturation of the chromoprotein. In the C-terminal domain, modification of only one cysteine residue (Cys-939) required protein denaturation. Since all 22 half cystines can be modified with iodoacetamide without reduction of the chromoprotein, it follows that oat phyA does not have any disulfide bonds. We found that Cys-311, Cys-774, Cys-961, and Cys-995 could be easily partially oxidized under the conditions used for phytochrome isolation. The surface topography/conformation of oat phyA and its role in protein-protein recognition in phytochrome-mediated signal transduction are discussed in terms of the relative reactivity of cysteine residues. PMID- 9730826 TI - Structure-function relationships of human liver cytochromes P450 3A: aflatoxin B1 metabolism as a probe. AB - Cytochromes P450 3A4 and 3A5, the dominant drug-metabolizing enzymes in the human liver, share >85% primary amino acid sequence identity yet exhibit different regioselectivity toward aflatoxin B1 (AFB1) biotransformation [Gillam et al., (1995) Arch. Biochem. Biophys. 317, 374-384]. P450 3A4 apparently prefers AFB1 3alpha-hydroxylation, which results in detoxification and subsequent elimination of the hepatotoxin, over AFB1 exo-8,9-oxidation. In contrast, P450 3A5 is incapable of appreciable AFB1 3alpha-hydroxylation and converts it predominantly to the exo-8,9-oxide which is genotoxic. To elucidate the structural features that govern the regioselectivity of the human liver 3A enzymes in AFB1 metabolism and bioactivation, a combination of approaches including sequence alignment, homology modeling, and site-directed mutagenesis was employed. Specifically, the switch in AFB1 regioselectivity was examined after individual substitution of the divergent amino acids in each of the six putative substrate recognition sites (SRSs) of P450 3A4 with the corresponding amino acid of P450 3A5. Of the P450 3A4 mutants examined, P107S, F108L, N206S, L210F, V376T, S478D, and L479T mutations resulted in a significant switch of P450 3A4 regioselectivity toward that of P450 3A5. The results confirmed the importance of some of these residues in substrate contact in the active site, with residue N206 (SRS-2) being critical for AFB1 detoxification via 3alpha-hydroxylation. Moreover, the P450 3A4 mutant N206S most closely mimicked P450 3A5, not only in its regioselectivity of AFB1 metabolism but also in its overall functional capacity. Furthermore, the other SRS-2 mutant, L210F, also resembled P450 3A5 in its overall AFB1 metabolism and regioselectivity. These findings reveal that a single P450 3A5 SRS domain (SRS-2) is capable of conferring the P450 3A5 phenotype on P450 3A4. In addition, some of these P450 3A4 mutations that affected AFB1 regioselectivity had little influence on testosterone 6beta-hydroxylation, thereby confirming that each substrate-P450 active site fit is indeed unique. PMID- 9730827 TI - Molecular, enzymatic, and regulatory characterization of rat kidney cytochromes P450 4A2 and 4A3. AB - The cDNAs encoding cytochromes CYP 4A2 and 4A3 were cloned by RT-PCR amplification of male rat kidney and liver RNAs, respectively. Sequence analysis demonstrated that these cDNAs were nearly identical to the published sequences for CYPs 4A2 and 4A3. CYP 4A2 and 4A3 share extensive sequence homology that extends into their 3'- and 5'-untranslated segments ( approximately 97% overall nucleotide identity). Analysis of cDNA and genomic DNA sequences shows that a sequence of 123 bp, recognized as an intron during the processing of CYP 4A2 transcripts, is conserved in the 4A3 mRNAs and that these otherwise highly homologous genes show different exon-intron distributions. The CYP 4A2 and 4A3 cDNAs were expressed in a baculovirus-insect cell expression system. Purified recombinant CYP 4A2 oxidized arachidonic acid to a mixture of 19- and 20 hydroxyeicosatetraenoic acids (20 and 80% of the total products, respectively). Reaction rates were maximal when CYP 4A2 was reconstituted in the presence of an equimolar concentration of cytochrome b5 and a 10-fold molar excess of NADPH cytochrome P450 reductase. Studies using microsomal fractions isolated from noninfected insect cells and from cells infected with CYP 4A3 recombinant baculoviruses showed (a) the presence of an endogenous lauric acid omega hydroxylase and arachidonic acid epoxygenase in the noninfected cells, (b) the CYP 4A3-dependent oxidation of lauric acid to 11- and 12-hydroxylaurate (24 and 76% of the total products, respectively), and (c) the lack of arachidonic acid metabolism by microsomal recombinant CYP 4A3. Nucleic acid hybridization and immunoelectrophoresis studies demonstrated that (a) CYP 4A2 transcripts are abundantly expressed in the female kidney and that CYP 4A3 is expressed in female but not in male liver, (b) anti-CYP 4A2 immunoreactive material was detected only in the male kidney, (c) male and female livers or kidneys support only low levels of CYP 4A3 translation, and (d) excess dietary salt does not alter the kidney levels of mRNA transcripts encoding CYP 4A1, 4A2, or 4A3 or change the levels of microsomal anti-4A1 or -4A2 immunoreactive proteins. Finally, no significant differences were observed between Dahl salt resistant or Dahl salt sensitive rats in the levels and/or salt regulation of mRNA transcripts enecoding CYP 4A1, 4A2, or 4A3 or the in levels of the corresponding proteins. PMID- 9730828 TI - Mechanism of Rab geranylgeranylation: formation of the catalytic ternary complex. AB - Rab proteins are geranylgeranylated on one or two C-terminal cysteines by Rab geranylgeranyl transferase (RabGGTase). The reaction is dependent on a Rab binding protein, termed Rab escort protein (REP). Here, we studied the role of REP in the geranylgeranylation reaction. We first characterized the interaction between REP and ungeranylgeranylated Rab using analytical ultracentrifugation and a fluorescence-based assay. We measured an equilibrium dissociation constant of 0.2 microM for the formation of a 1:1 REP-Rab complex and showed that this interaction relies mostly on ionic bonds and does not involve the two C-terminal cysteine residues. Second, we show that REP is required for recognition of Rab by RabGGTase and therefore that the REP-Rab complex is the true substrate for RabGGTase. Third, we show that free REP inhibits the geranylgeranylation reaction, suggesting that the complex is recognized by RabGGTase primarily via a REP-binding site. Our data suggest a model whereby REP behaves kinetically as an essential activator of the reaction. PMID- 9730829 TI - MgATP-dependent and MgATP-independent [3H]noradrenaline release from perforated synaptosomes both use N-ethylmaleimide-sensitive fusion protein. AB - In streptolysin-O (SLO)-perforated rat brain cortical synaptosomes, Ca2+-induced [3H]noradrenaline (3H-NA) release began with a phase lasting about 1 min that did not depend on MgATP. Subsequent release became increasingly MgATP-dependent. The first phase involved release from previously "primed" synaptic vesicles. MgATP dependent release, on the other hand, was release from unprimed vesicles that needed to be primed by ATP hydrolysis before they could be fused with the presynaptic membrane. Vesicle depriming was detected by observing that the initial release decreased when the synaptosomes were perforated and incubated for 2 min in the absence of MgATP before increasing Ca2+ to promote release. One millimolar N-ethylmaleimide (NEM) inhibited both MgATP-dependent and MgATP independent release at all times of incubation (0.5-5 min), and inhibition by NEM was partially reversed at short (0.5 min) and longer (5 min) times by adding intact N-ethylmaleimide sensitive fusion protein (NSF) to the perforated synaptosomes. Polyclonal antibodies against the N-terminal domain of NSF produced dose-dependent inhibition of Ca2+-induced 3H-NA release. This inhibition occurred in both early and late release phases and was highly significant at early times if the perforated synaptosomes were preincubated for 2 min with anti-NSF. These results indicate participation of NSF both after vesicular fusion, probably for separation of SNARE proteins in v/t-SNARE complexes before endocytosis, and, surprisingly, after docking, possibly to maintain vesicles in a primed state and reverse depriming during regulated secretion. PMID- 9730830 TI - Catalytic significance of the specificity of divalent cations as KS* and kcat* cofactors for secreted phospholipase A2. AB - Calcium is required for the substrate binding and for the chemical step of the interfacial catalytic turnover cycle of pancreatic phospholipase A2 (PLA2), but not for the binding of the enzyme to the interface. The role of calcium and other divalent cations (C) is analyzed for the effect on the substrate binding and kcat* for the chemical step. The cofactor role of 3d-cations(II) (C) for the hydrolysis of dimyristoylphosphatidylmethanol (DMPM) vesicles is characterized as an equilibrium dissociation constant for the interfacial binary (E*C) and ternary (E*CL) complexes of PLA2 and substrate mimics (L). Of the cations(II) that promote the binding of a mimic to the enzyme at the interface (E*), only a subgroup supports the chemical step. For example, Cd, Zn, and Cu form ternary E*CL complexes with kcat* of <1 s-1, compared to the rate of >100 s-1 with Ca, Fe, Mn, Co, and Ni. Oxygen exchange from H218O to the products of hydrolysis of DMPM incorporates one 18O in myristate. Incorporation of the first and second 18O occurs during the incubation of both the products of hydrolysis in H218O with PLA2 and Ca, but not with Zn. The cation-dependent changes in the UV difference spectrum, associated with the formation of E*C and E*CL, suggest that the changes are mainly due to catalytic His-48, and possibly Tyr-52 and Tyr-73, and are different with Ca as opposed to Zn. These results and simulations suggest considerable plasticity in the calcium binding and catalytic site environment. It is proposed that the higher ground state stability of the E*CS complex with the inhibitory cations increases the effective activation energy. For the chemical step, calcium coordinated with a nucleophilic water and the ester carbonyl oxygen facilitates the near-attack geometry in the E*CaS, and the His-48.Asp-99 pair acts as a proton acceptor. As a prelude to establishing the catalytic mechanism, factors controlling the energetically demanding transition state are also discussed. PMID- 9730831 TI - Electrostatic and hydrophobic contributions to the folding mechanism of apocytochrome c driven by the interaction with lipid. AB - In aqueous solution, while cytochrome c is a stably folded protein with a tightly packed structure at the secondary and tertiary levels, its heme-free precursor, apocytochrome c, shows all features of a structureless random coil. However, upon interaction with phospholipid vesicles or lysophospholipid micelles, apocytochrome c undergoes a conformational transition from its random coil in solution to an alpha-helical structure on association with lipid. The driving forces of this lipid-induced folding process of apocytochrome c were investigated for the interaction with various phospholipids and lysophospholipids. Binding of apocytochrome c to negatively charged phospholipid vesicles induced a partially folded state with approximately 85% of the alpha-helical structure of cytochrome c in solution. In contrast, in the presence of zwitterionic phospholipid vesicles, apocytochrome c remains a random coil, suggesting that negatively charged phospholipid headgroups play an important role in the mechanism of lipid induced folding of apocytochrome c. However, negatively charged lysophospholipid micelles induce a higher content of alpha-helical structure than equivalent negatively charged diacylphospholipids in bilayers, reaching 100% of the alpha helix content of cytochrome c in solution. Furthermore, micelles of lysolipids with the same zwitterionic headgroup of phospholipid bilayer vesicles induce approximately 60% of the alpha-helix content of cytochrome c in solution. On the basis of these results, we propose a mechanism for the folding of apocytochrome c induced by the interaction with lipid, which accounts for both electrostatic and hydrophobic contributions. Electrostatic lipid-protein interactions appear to direct the polypeptide to the micelle or vesicle surface and to induce an early partially folded state on the membrane surface. Hydrophobic interactions between nonpolar residues in the protein and the hydrophobic core of the lipid bilayer stabilize and extend the secondary structure upon membrane insertion. PMID- 9730832 TI - Oxidative decarboxylation of 6-phosphogluconate by 6-phosphogluconate dehydrogenase proceeds by a stepwise mechanism with NADP and APADP as oxidants. AB - Primary kinetic deuterium, 13C, and multiple deuterium/13C-isotope effects on V/K6PG have been measured for the Candida utilis (cu) and sheep liver (sl) 6 phosphogluconate dehydrogenases (6PGDH). With NADP as the dinucleotide substrate, the following values of D(V/K6PG), 13(V/K6PG)H, and 13(V/K6PG)D were measured at pH 8 for cu6PGDH (sl6PGDH): 1.57 +/- 0.08 (1.87 +/- 0.10), 1.0209 +/- 0.0005 (1.0059 +/- 0.000 10), 1.0158 +/- 0.0001 (1.0036 +/- 0.0008). With APADP as the dinucleotide substrate, values for the above isotope effects at pH 8 are as follows: 2.98 +/- 0.08 (2.47 +/- 0.06), 1. 0106 +/- 0.0002 (1.0086 +/- 0.000 09), and 0.9934 +/- 0.0003 (0.9950 +/- 0.0003). Results indicate the oxidative decarboxylation of 6PG to the 1,2-enediol of ribulose 5-phosphate proceeds via a stepwise mechanism with hydride transfer preceding decarboxylation in all cases. The inverse 13C-isotope effect observed with APADP and 6PG-3d may reflect a preequlibrium isotope effect on the binding of 6PG preceding hydride transfer. Deuterium-isotope effects on V, V/KNADP, and V/K6PG are identical at all pHs and for both enzymes. The primary deuterium-isotope effect on V/K6PG for both enzymes is constant at pH values below the pK in the pH profile for V/K6PG, and decreases as the pH increases. Data suggest the development of rate limitation by a step or steps other than the hydride-transfer step as the pH is increased. PMID- 9730833 TI - A buried polar interaction can direct the relative orientation of helices in a coiled coil. AB - Coiled coils consist of bundles of two or more alpha-helices that are aligned in a parallel or an antiparallel relative orientation. The designed peptides, Acid p1 and Base-p1, associate in solution to form a parallel, heterodimeric two stranded coiled coil [O'Shea, E. K., Lumb, K. J., and Kim, P. S. (1993) Curr. Biol. 3, 658]. The buried interface of this complex is formed by hydrophobic Leu residues, with the exception of an Asn residue from each strand that is positioned to engage in a buried polar interaction. Substitution of these buried Asn residues by Leu residues results in a loss of structural uniqueness, as evidenced by a lack of a particular helix orientation in the Acid-Base coiled coil complex [Lumb, K. J., and Kim, P. S. (1995) Biochemistry 34, 8642]. Here, we alter the positions of the Asn residues in the Acid and Base peptides such that a buried polar interaction is only expected to occur when the helices are in an antiparallel orientation. The resulting peptides, Acid-a1 and Base-a1, associate to form a helical heterodimer, as shown by circular dichroism (CD) and equilibrium sedimentation centrifugation. The helix orientation preference has been measured using covalently linked, disulfide-containing heterodimers in which the constituent peptides are constrained to interact in either a parallel or an antiparallel orientation. Although both the parallel and antiparallel heterodimers form stable, helical structures, the antiparallel heterodimer is the predominant species at equilibrium when the heterodimers are allowed to undergo thiol-disulfide exchange. In addition, the antiparallel heterodimer is more stable to chemical denaturation than the parallel counterpart by approximately 2.3 kcal/mol. These results demonstrate that a single buried polar interaction in the interface between the helices of a coiled coil is sufficient to determine the relative orientation of its constituent helices. PMID- 9730834 TI - Catalytic and biophysical properties of a nitrogenase Apo-MoFe protein produced by a nifB-deletion mutant of Azotobacter vinelandii. AB - A Zn-immobilized metal-affinity chromatography technique was used to purify a poly-histidine-tagged, FeMo-cofactorless MoFe protein (apo-MoFe protein) from a nifB-deletion mutant of Azotobacter vinelandii. Apo-MoFe protein prepared in this way was obtained in sufficient concentrations for detailed catalytic, kinetic, and spectroscopic analyses. Metal analysis and electron paramagnetic resonance spectroscopy (EPR) were used to show that the apo-MoFe protein does not contain FeMo-cofactor. The EPR of the as-isolated apo-MoFe protein is featureless except for a minor S = 1/2 signal probably arising from the presence of either a damaged P cluster or a P cluster precursor. The apo-MoFe protein has an alpha2beta2 subunit composition and can be activated to 80% of the theoretical MoFe protein value by the addition of isolated FeMo-cofactor. Oxidation of the as-isolated apo MoFe protein by indigodisulfonate was used to elicit the parallel mode EPR signal indicative of the two-electron oxidized form of the P cluster (P2+). The midpoint potential of the PN/P2+ redox couple for the apo-MoFe protein was shown to be shifted by -63 mV when compared to the same redox couple for the intact MoFe protein. Although the apo-MoFe protein is not able to catalyze the reduction of substrates under turnover conditions, it does support the hydrolysis of MgATP at 60% of the rate supported by the MoFe protein when incubated in the presence of Fe protein. The ability of the apo-MoFe protein to specifically interact with the Fe protein was also shown by stopped-flow techniques and by formation of an apo MoFe protein-Fe protein complex. Finally, the two-electron oxidized form of the apo-MoFe protein could be reduced to the one-electron oxidized form (P1+) in a reaction that required Fe protein and MgATP. These results are interpreted to indicate that the apo-MoFe protein produced in a nifB-deficient genetic background [corrected] contains intact P clusters and P cluster polypeptide environments. Small changes in the electronic properties of P clusters contained within the apo-MoFe protein are most likely caused by slight perturbations in their polypeptide environments. PMID- 9730835 TI - A second magnesium ion is critical for ATP binding in the kinase domain of the oncoprotein v-Fps. AB - The activity of the kinase domain of the oncoprotein v-Fps was found to be sensitive to the concentration of magnesium ions. Plots of initial velocity versus free magnesium concentration are hyperbolic and do not extrapolate to the origin at stoichiometric ATP-Mg, indicating that there are two sites for metal chelation on the enzyme and the second is nonessential for catalysis. The second metal is strongly activating and increases the reaction rate constant almost 20 fold from 0.5 to 8.3 s-1 using 0.2 mM ATP-Mg and 1 mM peptide, EAEIYEAIE. This increase in rate is due to a large increase in the apparent affinity of ATP-Mg at high magnesium concentrations. At 0.5 and 10 mM free Mg2+, KATP-Mg is 3.6 and 0.22 mM, respectively. Extrapolation of the observed affinity of ATP-Mg to zero and infinite free metal indicates that KATP-Mg is greater than 8 mM in the absence of the second metal and 0.1 mM in the presence of the second metal, a minimum 80-fold enhancement. By comparison, free levels of the divalent ion do not influence maximum turnover (kcat) and have only a 2-fold effect on the Km for the peptide substrate between 0.5 and 20 mM free Mg2+. Viscosometric studies indicate that free Mg2+ does not influence the rates of phosphoryl transfer or net product release above 0.5 mM but does affect directly the dissociation constant for ATP-Mg. The Kd for ATP-Mg in the absence and presence of the second metal ion is >32 and 0.4 mM, respectively. At high magnesium concentrations, ATP Mg and the peptide substrate bind independently, while at lower concentrations (0.5 mM), there is significant negative binding synergism suggesting that the second metal may help to reduce charge repulsion between ATP-Mg and the peptide. The data indicate that the first metal is sufficient for phosphoryl transfer. While the second metal could have some influence on phosphoryl transfer or product binding, it is a potent activator that functions minimally by controlling ATP-Mg binding. PMID- 9730836 TI - A refined kinetic analysis of plasminogen activation by recombinant bovine tissue type plasminogen activator indicates two interconvertible activator forms. AB - Bovine tissue-type plasminogen activator (tPA) was heterologously expressed in the methylotrophic yeast Pichia pastoris and characterized structurally and kinetically. The bovine single-chain tPA-mediated activation of bovine plasminogen was studied in the presence and absence of fibrinogen fragments. We have proposed a refined new method of kinetic analysis which allows examination of both stationary and prestationary phases of this process. The investigation revealed the presence of two interconvertible forms of the recombinant bovine tPA being in equilibrium at a 1 to 50 ratio. Only the minor form was able to bind and activate plasminogen. Saturation of the whole pool of tPA required high plasminogen concentration (Km >/= 5 microM) in order to reverse the equilibrium between the two forms. Fibrinogen fragments activated the single-chain tPA due to preferential binding and stabilization of the minor "active" form of the enzyme until all the molecules of tPA were converted. The same mechanism could be applied to human tPA as well. The Km values, obtained for recombinant bovine and human tPA in the presence of fibrinogen fragments, were found to be similar (Km = 0.1 microM) while kcat of human tPA was 5-10 times higher. PMID- 9730837 TI - Stabilization of ribonuclease HI from Thermus thermophilus HB8 by the spontaneous formation of an intramolecular disulfide bond. AB - To identify factors that contribute to the thermal stability of ribonuclease HI (RNase HI) from Thermus thermophilus HB8, protein variants with a series of carboxyl-terminal truncations and Cys --> Ala mutations were constructed, and their thermal denaturations were analyzed by CD. The results indicate that Cys41 and Cys149 contribute to the protein stability, probably through the formation of a disulfide bond. Peptide mapping analysis for the mutant protein with only two cysteine residues, at positions 41 and 149, indicated that this disulfide bond is partially formed in a protein purified from Escherichia coli in the absence of a reducing reagent but is fully formed in a thermally denatured protein. These results suggest that the thermal stability of T. thermophilus RNase HI, determined in the absence of a reducing reagent, reflects that of an oxidized form of the protein. Comparison of the thermal stabilities and the enzymatic activities of the wild-type and truncated proteins, determined in the presence and absence of a reducing reagent, indicates that the formation of this disulfide bond increases the thermal stability of the protein by 6-7 degreesC in Tm and approximately 3 kcal/mol in DeltaG without seriously affecting the enzymatic activity. Since T. thermophilus RNase HI is present in a reducing environment in cells, this disulfide bond probably is not formed in vivo but is spontaneously formed in vitro in the absence of a reducing reagent. PMID- 9730838 TI - The mechanism of adenosylmethionine-dependent activation of methionine synthase: a rapid kinetic analysis of intermediates in reductive methylation of Cob(II)alamin enzyme. AB - Cobalamin-dependent methionine synthase catalyzes the transfer of a methyl group from methyltetrahydrofolate to homocysteine, generating tetrahydrofolate and methionine. During this primary turnover cycle, the enzyme alternates between the active methylcobalamin and cob(I)alamin forms of the enzyme. Formation of the cob(II)alamin prosthetic group by oxidation of cob(I)alamin or photolysis of methylcobalamin renders the enzyme inactive. Methionine synthase from E. coli catalyzes its own reactivation by a reductive methylation that involves electron transfer from reduced flavodoxin and methyl transfer from AdoMet. This process has been proposed to involve formation of a transient cob(I)alamin intermediate that is then trapped by methyl transfer from AdoMet. During aerobic growth of E. coli, electrons for this process are ultimately derived from NADPH, and electron transfer does not generate a detectable level of cob(I)alamin due to the large potential difference between the NADPH/NADP+ couple and the cob(I)alamin/cob(II)alamin couple. In this paper, we show that even in the presence of the strong reductant flavodoxin hydroquinone, cob(I)alamin is not observed as a significant intermediate. We demonstrate, however, that this is due to a rate-limiting reorganization of the cobalt ligand environment from five coordinate to four-coordinate cob(II)alamin. Mutation of aspartate 757 to glutamate results in a cob(II)alamin enzyme that is approximately 70% four coordinate, and reductive methylation of this enzyme using flavodoxin hydroquinone as the electron donor proceeds through a kinetically competent cob(I)alamin intermediate. Furthermore, wild-type cob(I)alamin enzyme produced by chemical reduction reacts with AdoMet in a kinetically competent reaction. We provide evidence that methyl transfer from AdoMet to cob(I)alamin enzyme results initially in formation of a five-coordinate methylcobalamin enzyme that slowly decays to the active six-coordinate methylcobalamin enzyme. We propose a kinetic scheme for reductive methylation of wild-type cob(II)alamin enzyme by adenosylmethionine and flavodoxin hydroquinone in which slow conformational changes mask the relatively fast electron and methyl transfer steps. PMID- 9730839 TI - Thermodynamics of ligand binding and catalysis in human liver medium-chain acyl CoA dehydrogenase: comparative studies involving normal and 3'-dephosphorylated C8-CoAs and wild-type and Asn191 --> Ala (N191A) mutant enzymes. AB - Following our demonstration that the terminal 3'-phosphate group of acyl-CoA substrates (which is confined to the exterior of the protein structure, and is fully exposed to the outside solvent environment) exhibits a functional role in the recombinant human liver medium-chain acyl-CoA dehydrogenase (MCAD)-catalyzed reaction [Peterson, K. L., and Srivastava, D. K. (1997) Biochem. J. 325, 751 760], we became interested in delineating its thermodynamic contribution in stabilizing the "ground" and "transition" state structures during enzyme catalysis. Since the 3'-phosphate group of the coenzyme A thiolester has the potential to form a hydrogen bond with the side chain group of Asn-191, these studies were performed utilizing both normal and 3'-dephosphorylated forms of octanoyl-CoA and octenoyl-CoA (cumulatively referred to as C8-CoA) as the physiological substrate and product of the enzyme, respectively, as well as utilizing wild-type and Asn191 --> Ala (N191A) site-specific mutant enzymes. The experimental data revealed that the enthalpic contribution of the 3'-phosphate group was similar in both ground and transition states, and was primarily derived from the London-van der Waals interactions (between the 3'-phosphate group of C8 CoA and the surrounding protein moiety), rather than from the potential hydrogen bonding. The temperature dependence of DeltaH degrees for the binding of octenoyl CoA and 3'-dephosphooctenoyl-CoA revealed that the deletion of the 3'-phosphate group from octenoyl-CoA increased the magnitude of the heat capacity changes (DeltaCp degrees) from -0.53 to -0.59 kcal mol-1 K-1. Although the latter effect could be attributed to an increase in the relative hydrophobicity of the ligand, the experimentally observed DeltaCp degrees's (for either of the ligands) could not be predicted on the basis of the changes in the solvent-accessible surface areas of the enzyme and ligand species. These coupled with the fact that the DeltaCp degrees for the binding of octenoyl-CoA to pig kidney MCAD (which is believed to be structurally identical to human liver MCAD) is only -0.37 kcal mol 1 K-1 [Srivastava, D. K., Wang, S., and Peterson, K. L. (1997) Biochemistry 36, 6359-6366] prompt us to question the reliability of predicting the DeltaCp degrees values of the enzyme-ligand complexes from their X-ray crystallographic data. Arguments are presented that certain intrinisic limitations of the crystallographic data preclude kinetic and thermodynamic predictions about the enzyme-ligand complexes and enzyme catalysis. PMID- 9730841 TI - Prolonged stability of brain natriuretic peptide: importance for non-invasive assessment of cardiac function in clinical practice. AB - 1.BNP and ANP are important research indices of severity of heart failure. However, uncertainty regarding the stability of these peptides at room temperature has limited their use to assess cardiac function in routine clinical practice. 2. We assessed the stability of BNP and ANP in blood samples left for 2 h or 2 days at room temperature compared with levels in blood processed immediately (initial). These times were chosen to reflect possible times for samples to be processed in a hospital outpatient clinic (2 h) or a blood sample posted to a laboratory from general practice (2 days). Samples were obtained from eight heart transplant recipients. Blood was separated and plasma stored immediately after collection (initial) and after 2 h or 2 days at room temperature respectively. 3. Initial plasma BNP and ANP values measured by radioimmunoassay after Sep-Pak extraction were 38.9+/-11.1(S.E.M.) pg/ml and 113.6+/-28.1 pg/ml, respectively. After 2 h at room temperature there was no significant fall in either peptide level (35.5+/-9.9 pg/ml, BNP; 104. 9+/-30.6 pg/ml, ANP). However, after 2 days at room temperature there was a significant fall in ANP to 38.1+/-12.6 pg/ml (P<0.005 versus initial level). In contrast, there was no significant fall in BNP after 2 days (32.0+/-8.4 pg/ml). After 2 days at room temperature only 30.4+/-4.3% of the ANP remained, but 86.0+/-5.0% of BNP compared with the initial ANP and BNP measurements. 4. Our study clearly showed that ANP is stable for 2 h and thus could be useful as a screening test for heart disease in hospital. In contrast, BNP remained stable for 2 days. Measuring BNP may thus be practical as a test of heart function both for routine use in hospital and by general practitioners in the community. PMID- 9730840 TI - Functional comparison of two human monocyte chemotactic protein-2 isoforms, role of the amino-terminal pyroglutamic acid and processing by CD26/dipeptidyl peptidase IV. AB - Human Monocyte Chemotactic Protein (MCP)-2 has originally been isolated from stimulated osteosarcoma cells as a chemokine coproduced with MCP-1 and MCP-3. Here, a 5'-end extended MCP-2 cDNA was cloned from a human testis cDNA library. It encoded a 76 residue MCP-2 protein, but differed from the reported bone marrow derived MCP-2 cDNA sequence in codon 46, which coded for a Lys instead of a Gln. This MCP-2Lys46 variant, caused by a single nucleotide polymorphism (SNP), was biologically compared with MCP-2Gln46. The coding regions were subcloned into the bacterial expression vector pHEN1, and after transformation of Escherichia coli, the two MCP-2 protein variants were recovered from the periplasm. The recombinant proteins were purified to homogeneity by heparin-Sepharose affinity chromatography and reversed-phase HPLC. Edman degradation revealed a Gln residue at the NH2 terminus instead of a pGlu. To evaluate the influence of the cyclization, this Gln was chemically converted into pGlu in both MCP-2 variants. The conversion was confirmed by electrospray mass spectrometry. rMCP-2Gln46 and rMCP-2Lys46 and the NH2-terminal cyclic counterparts were tested on monocytic cells in calcium mobilization and chemotaxis assays. No significant difference in biological activity was observed between the rMCP-2Gln46 and rMCP-2Lys46 isoforms. However, for both MCP-2 variants the NH2-terminal pyroglutamate was shown to be essential for chemotaxis, but not for calcium mobilization. NH2 terminal truncation of rMCP-2Lys46 by the serine protease CD26/dipeptidyl peptidase IV (CD26/DPP IV) resulted in the cleavage of the NH2-terminal Gln-Pro dipeptide, whereas synthetic MCP-2 with an amino-terminal pGlu remained unaffected. CD26/DPP IV-clipped rMCP-2Lys46(3-76) was almost completely inactive in both chemotaxis and signaling assays. These observations indicate that the NH2 terminal pGlu in MCP-2 is necessary for chemotactic activity but also that it protects the protein against degradation by CD26/DPP IV. PMID- 9730842 TI - A central gamma-aminobutyric acid mechanism in cardiac vagal control in man revealed by studies with intravenous midazolam. AB - 1. Animal studies show that cardiac vagal tone can be modified by gamma aminobutyric acid neurons acting at several sites in the central nervous system. The present study has attempted to determine whether similar control exists in humans by using midazolam, a benzodiazepine. Benzodiazepines exert their main actions on the central nervous system by interacting co-operatively at the gamma aminobutyric acid receptor. 2. Twenty patients took part in the study before undergoing cardiac catheterization. After resting for 20 min in a semi-supine position on a couch, ECG, blood pressure and respiration were recorded for 5-min periods with either controlled (fixed) or free respiration. During this time a baroreceptor sensitivity test was conducted. 3. Doses of 1 mg and 5 mg of midazolam were administered intravenously. 4. Five-minute segments of data, before and after midazolam, were subjected to power spectral and time-domain analysis. 5. Midazolam caused a decrease in the high-frequency and an increase in the low-frequency components of the power spectral density plot, and in addition reduced the mean R-R interval and R-R variability expressed as the interquartile difference, and pNN50. There were no significant changes in the sensitivity of the baroreflex or in the systolic, diastolic and average blood pressures. 6. This decrease in variability of heart period, particularly at a controlled respiratory frequency, strongly suggests that cardiac vagal tone in man can be regulated by gamma-aminobutyric acid neurons. PMID- 9730843 TI - Lack of habituation of the pattern of cardiovascular response evoked by sound in subjects with primary Raynaud's disease. AB - 1. The vasospasm of primary Raynaud's disease can be triggered by acute emotional stress. We have studied the pattern of cardiovascular response evoked by acute emotional stress, a sound stimulus of 90 dB, 2 kHz for 30 s, in eight subjects with primary Raynaud's disease and in eight age- and sex-matched controls, the sound being repeated five times on each of days 1, 3 and 5. 2. In controls, the first sound evoked the pattern of the alerting response that is characteristic of acute emotional stress: a rise in arterial pressure and heart rate, a decrease in vascular conductance in the cutaneous circulation of the digit, assessed by laser Doppler recording of erythrocyte (red cell) flux in the digit divided by arterial pressure, and an increase in forearm muscle vascular conductance, assessed from forearm blood flow recorded by venous occlusion plethysmography divided by arterial pressure. 3. In the subjects with primary Raynaud's disease, baselines of arterial pressure, digital cutaneous vascular conductance and forearm vascular conductance were not significantly different from those of the controls and they too showed the alerting response to the first sound, the magnitudes of the changes being comparable to those of the controls. 4. In both the controls and subjects with primary Raynaud's disease, the evoked responses were consistent on repetition of the sound on day 1. In contrast, judging from the means of the changes evoked on each day, the controls showed habituation of the individual components of the alerting response over days 1, 3 and 5, whereas the subjects with primary Raynaud's disease showed no habituation of either the forearm muscle vasodilatation or the digital vasoconstriction. Conversely, the decrease in digital cutaneous vascular conductance evoked by a single deep breath was fully reproducible in both controls and subjects with primary Raynaud's disease when tested at the beginning and end of each experimental day. 5. These results allow the novel conclusion that subjects with primary Raynaud's disease have an abnormality of the central neural modulation of the brain stem areas that integrate the cardiovascular components of the alerting response to acute emotional stress, such that habituation of the vasodilator and vasoconstrictor components of the response on repetition of the stimulus is impaired. We propose that such persistence of vasoconstrictor responses to stressful stimuli predisposes to vasospasm, particularly if neurally mediated vasoconstriction is reinforced by locally released vasoconstrictor factors. PMID- 9730844 TI - Factors determining cardiac hypertrophy in hypertensive patients with or without peripheral vascular disease. AB - 1. Coronary ischaemic disease and congestive heart failure are the principal causes of mortality in patients with peripheral vascular disease. Whether cardiac hypertrophy is present and even more pronounced in peripheral vascular disease than in other populations has never been explored.2.Twenty-five hypertensive patients were investigated, 11 without and 14 with peripheral vascular disease, matched for age, sex, mean arterial pressure and antihypertensive drug treatment. Cardiac mass was determined using echocardiography together with measurement of systemic blood pressure, ratio between ankle systolic pressure (ASP) and brachial systolic pressure (BSP), and standard biochemical parameters including natriuresis per 24 h.3. At the same mean arterial pressure, patients with peripheral vascular disease had a significantly higher cardiac mass (157+/-12 versus 116+/-6 g/m2; P<0.01), pulse pressure (81+/-5 versus 55+/-4 mmHg; P<0.01) and natriuresis (180+/-17 versus 144+/-6 mmol/24h; P<0. 01) than controls. Using univariate correlations, cardiac mass was positively associated with pulse pressure, mean arterial pressure and natriuresis, and negatively with the ASP/BSP ratio. On the basis of multivariate regression analysis, only natriuresis was positively correlated to cardiac mass.4. Patients with peripheral vascular disease develop a higher degree of cardiac hypertrophy in comparison with hypertensive subjects with the same level of mean arterial pressure. Sodium intake rather than mechanical factors seems to be the major modulating factor which influences the degree of cardiac hypertrophy. PMID- 9730846 TI - Respiratory correlates of muscle sympathetic nerve activity in heart failure. AB - 1. Sympathetic activation in congestive heart failure indicates a poor prognosis. Haemodynamic correlates of increased sympathetic nerve traffic to muscle (MSNA) and to the heart have been well characterized, but these account for only 50 to 60% of the variance in sympathetic activity between patients.2. In healthy subjects, breathing pattern modulates MSNA and positive airway pressure consistently increases MSNA. However, in patients with heart failure, the influence of spontaneous breathing pattern and of short-term application of nasal continuous positive airway pressure on MSNA have not been described.3. Spontaneous breathing frequency, tidal volume, end-expiratory lung volume, PCO2 and MSNA were recorded, along with blood pressure, heart rate and stroke volume in 14 men with congestive heart failure of idiopathic or ischaemic origin (left ventricular ejection fraction <35%). Measurements were made during baseline rest, followed by 45 min of either nasal continuous positive airway pressure applied at 10 cmH2O (n=9), or spontaneous breathing, in the absence of nasal continuous positive airway pressure (time control; n=6).4. At baseline, there was a significant positive correlation between MSNA burst frequency and breathing frequency (r=0.758, P=0.001), and an inverse correlation between MSNA burst incidence and tidal volume (r=-0.705, P=0.005). These relationships were independent of left ventricular ejection fraction, stroke volume or cardiac output.5.Nasal continuous positive airway pressure increased end-expiratory lung volume, but had no effect on breathing frequency, tidal volume or MSNA.6. In patients with congestive heart failure, there is a significant independent and previously unrecognized correlation between spontaneous breathing pattern and MSNA; patients with rapid shallow breathing exhibit the highest degree of sympathetic activation. In distinct contrast to healthy subjects, the short-term application of nasal continuous positive airway pressure at 10 cmH2O does not increase MSNA in congestive heart failure. PMID- 9730845 TI - Human cardiovascular variability, baroreflex and hormonal adaptations to a blood donation. AB - 1. We studied cardiovascular variability, baroreflex and blood volume regulating hormones to determine the relative roles of autonomic regulation and hormones during blood donation.2. The sympathetic response was studied by measuring the R R interval and systolic blood pressure variability using coarse graining spectral analysis in eight blood donors. Beat-by-beat R-R intervals and blood pressure were recorded for 20 min before and 5 min after a whole-blood donation of 480+/ 10 ml (about 7 ml/kg of blood volume, over 4 min). Plasma catecholamines, vasopressin, atrial natriuretic peptide, endothelin, active renin, osmolality, Na+, K+, haemoglobin and haematocrit were measured just before and after blood withdrawal.3. Blood donation led to increases in the plasma catecholamines (adrenaline, 21+/-2 versus 35+/-3 pg/ml; noradrenaline, 229+/-26 versus 323+/-37 pg/ml; dopamine, 34+/-3 versus 66+/-9 pg/ml) and in systolic blood pressure (130+/-6 versus 140+/-5 mmHg). These changes were independent of ionic or slow endocrine mechanisms. Heart rate, cardiovascular variability and the spontaneous baroreflex sensitivity did not change despite the increase in blood pressure and catecholamines. Thus the peripheral vascular control was probably involved.4. We conclude that the absence of any change in heart rate usually observed during non hypotensive hypovolaemic stress is probably due to the sympathetic activation being counterbalanced by the high supine vagal tone at the heart and not to the heterogeneous nature of the sympathetic neural response or to changes in sympathetic and parasympathetic activity without any change in autonomic balance. PMID- 9730847 TI - Subjects with obstructive pulmonary disease tend to be chronically vasodilated. AB - 1. In 12 unselected outpatients with chronic obstructive pulmonary disease and six controls, arterial pH, PaO2, PaCO2 and oxygen saturation (SaO2), forced expiratory volume in 1.0 s (FEV1.0) and vital capacity were measured. Subjects were grouped into those with or without obstruction based on the Tiffenau index. The Baseline Dyspnoea Index was employed to objectify the severity of dyspnoea and the Borg index to evaluate the subjective sensation. Blood pressure was measured with a sphygmomanometer; calf arterial flow both at rest and during reactive hyperaemia with a plethysmograph. Basal and minimal resistance were calculated.2.FEV1.0 was 26% lower in patients with obstruction than in controls, and was also lower in patients with moderate-to-severe obstruction compared with those with mild or no obstruction. Arterial flow (75% greater in the patients with obstruction) progressively increased with increasing severity of obstruction, being 54% higher in those with mild obstruction than in those with no obstruction (P<0.001), and 28% higher in moderate-severe than in mild obstruction (P<0.005). In multiple regressions, F correlated inversely with FEV1.0, PaO2 and SaO2, and directly with PaCO2. Basal resistance correlated positively with FEV1.0, SaO2 and the Tiffenau index, and inversely with PaCO2 (r= 0.52, P=0.02). Minimal resistance was significantly lower in obstructed than in non-obstructed subjects. Both basal and minimal resistance progressively decreased, although insignificantly, with worsening bronchial obstruction. PaCO2 did not correlate with any haemodynamic parameter. Borg index correlated indirectly with FEV1.0 and basal resistance directly with arterial flow.3. Patients with chronic obstructive pulmonary disease therefore tend to show chronic vasodilatation depending on hypoxia rather than PaCO2. Other mechanisms could be involved in this phenomenon. The Borg index is a good indicator of oxygen desaturation and vasodilatation. PMID- 9730848 TI - Fractal dimension and approximate entropy of heart period and heart rate: awake versus sleep differences and methodological issues. AB - 1. Investigations that assess cardiac autonomic function include non-linear techniques such as fractal dimension and approximate entropy in addition to the common time and frequency domain measures of both heart period and heart rate. This article evaluates the differences in using heart rate versus heart period to estimate fractal dimensions and approximate entropies of these time series.2. Twenty-four-hour ECG was recorded in 23 normal subjects using Holter records. Time series of heart rate and heart period were analysed using fractal dimensions, approximate entropies and spectral analysis for the quantification of absolute and relative heart period variability in bands of ultra low (<0.0033 Hz), very low (0. 0033-0.04 Hz), low (0.04-0.15 Hz) and high (0.15-0.5 Hz) frequency.3. Linear detrending of the time series did not significantly change the fractal dimension or approximate entropy values. We found significant differences in the analyses using heart rate versus heart period between waking up and sleep conditions for fractal dimensions, approximate entropies and absolute spectral powers, especially for the power in the band of 0.0033-0.5 Hz. Log transformation of the data revealed identical fractal dimension values for both heart rate and heart period. Mean heart period correlated significantly better with fractal dimensions and approximate entropies of heart period than did corresponding heart rate measures.4. Studies using heart period measures should take the effect of mean heart period into account even for the analyses of fractal dimension and approximate entropy. As the sleep-awake differences in fractal dimensions and approximate entropies are different between heart rate and heart period, the results should be interpreted accordingly. PMID- 9730849 TI - Influences of carvedilol treatment on the effects of acetylcholine on regional haemodynamics in the spontaneously hypertensive rat. AB - 1. In a previous report, we have shown that vasodilatation induced by acetylcholine is impaired in the kidney and the heart of the spontaneously hypertensive rat (SHR) in vivo. The present investigation was performed to study the influence of oral antihypertensive treatment with carvedilol for 6 to 10 weeks on acetylcholine-induced changes in regional haemodynamics in SHR in vivo. Cardiac output, regional blood flow and vascular resistance in organs of major importance in hypertensive disease, such as the kidney, heart, skeletal muscle, brain and eye, were measured with radioactively labelled microspheres in anaesthetized rats (aged 12-16 weeks).2. Mean arterial blood pressure was significantly lower in the carvedilol-treated SHR group (156+/-3 mmHg, n=17) than in an untreated SHR group (172+/-6 mmHg, n=13). Infusion of acetylcholine (2 microgram.min-1.kg-1) caused similar significant reductions in blood pressure in the two groups (-13+/-1% and -14+/-2%). However, acetylcholine induced a significant increase in total peripheral vascular resistance in the carvedilol group (29+/-10%, P<0.01), whereas no significant change was observed in the control group (0+/-11%).3. Acetylcholine significantly increased renal vascular resistance in the carvedilol group (+62+/-15%, P<0.01), but did not change vascular resistance in the control group (-6+/-6%). In the heart, acetylcholine did not affect vascular resistance in the carvedilol group, but reduced vascular resistance significantly in the control group (-17+/-8%, P<0.05). The circulatory changes induced by acetylcholine in the skeletal muscle, brain and ophthalmic circulation did not differ between the groups.4. In conclusion, the results demonstrate that long-term oral carvedilol treatment in the SHR did not enhance acetylcholine-induced vasodilatation, but instead pronounced renal vasoconstriction was induced by acetylcholine, which could partly be due to a decreased cardiac index. PMID- 9730850 TI - Methylenetetrahydrofolate reductase polymorphism (C-677T) and coronary artery disease. AB - 1. Many studies have shown that hyperhomocysteinaemia is a risk factor for atherosclerotic vascular disease. A mutation (C-677T) in the gene coding for the methylenetetrahydrofolate reductase (MTHFR) enzyme has been shown to produce a thermolabile form of the enzyme. Homozygosity for this mutation has been correlated with an elevated plasma homocysteine concentration. The present study aimed to determine whether this mutation was a risk factor for coronary artery disease (CAD). This was achieved by comparing the frequency of the C-677T mutation in patients with angiographically proven CAD against angiographically normal patients in two separate U.K. samples. The analysis was repeated with CAD patients split into those with >=99% stenosis of arteries and those without, to establish whether the C-677T mutation could be correlated with severity of CAD.2. Two patient groups were selected from London and Sheffield. The London group comprised 174 cases and 148 controls. The Sheffield group comprised 93 cases and 85 controls. The DNA samples of the patients were genotyped by polymerase chain reaction and restriction enzyme digestion.3. For London the homozygous C-677T frequencies were: 0.07 (controls), 0.09 (CAD without >=99% stenosis) and 0.10 (CAD with >=99% stenosis). For Sheffield the homozygous C-677T frequencies were: 0.08 (controls), 0.10 (CAD without >=99% stenosis) and 0.11 (CAD with >=99% stenosis). No association was found between the C-677T mutation and CAD in our sample geographical groups. Statistical comparison by genotype distribution for 0 VD (no vessel disease, i.e. 0% diameter reduction in all epicardial arteries) versus CAD without >=99% stenosis: London, P=0.19; Sheffield, P=0.53; 0 VD versus CAD with >=99% stenosis: London, P=0. 23; Sheffield, P=0.55. PMID- 9730851 TI - Diminished insulin clearance during late pregnancy in patients with type I diabetes mellitus. AB - 1. Intensive insulin treatment of patients with Type I diabetes mellitus during pregnancy is associated with a high frequency of serious hypoglycaemic events. A potential change in insulin metabolism during pregnancy may affect both the frequency and the severity of insulin-induced hypoglycaemia.2. In 10 patients with Type I diabetes, during the third trimester of pregnancy and 5 to 13 months after delivery, hypoglycaemia was induced by the hyperinsulinaemic hypoglycaemic clamp technique. A constant high-dose intravenous insulin infusion was administered for 150 min and arterial blood glucose was clamped at 2.2 mmol/l by counterregulation with intravenous glucose. During the experiment venous samples were collected for later analysis of free plasma insulin, whereby the metabolic clearance rate of insulin could be calculated.3. The desired blood glucose level was approached after approximately 60 min of insulin infusion. After just 30 min the insulin levels were significantly higher during pregnancy compared with after delivery. In addition, the steady-state insulin level from 90 to 150 min was significantly higher during pregnancy.4. From the steady-state insulin levels at 90 to 150 min, the metabolic clearance rate of insulin was calculated, being 24% higher after delivery.5. We conclude that there is a decreased metabolic clearance rate of insulin during pregnancy. This might be due to altered blood flow distribution, decreased hepatic insulin extraction and relative increase in body fat during pregnancy. A decreased clearance of insulin will contribute to the risk for serious hypoglycaemic events in patients with Type I diabetes during pregnancy. PMID- 9730852 TI - Subcutaneous glucagon-like peptide-1 improves postprandial glycaemic control over a 3-week period in patients with early type 2 diabetes. AB - 1.Glucagon-like peptide-1 (7-36) amide (GLP-1) is released into the circulation after meals and is the most potent physiological insulinotropic hormone in man. GLP-1 has the advantages over other therapeutic agents for Type 2 diabetes of also suppressing glucagon secretion and delaying gastric emptying. One of the initial abnormalities of Type 2 diabetes is the loss of the first-phase insulin response, leading to postprandial hyperglycaemia.2. To investigate the therapeutic potential of GLP-1 in Type 2 diabetes, six patients were entered into a 6-week, double-blind crossover trial during which each received 3 weeks treatment with subcutaneous GLP-1 or saline, self-administered three times a day immediately before meals. A standard test meal was given at the beginning and end of each treatment period.3.GLP-1 reduced plasma glucose area under the curve (AUC) after the standard test meal by 58% (AUC, 0-240 min: GLP-1 start of treatment, 196+/-141 mmol.min-1.l-1; saline start of treatment, 469+/-124 mmol.min-1.l-1; F=16.4, P<0.05). The plasma insulin excursions were significantly higher with GLP-1 compared with saline over the initial postprandial 30 min, the time period during which the GLP-1 concentration was considerably elevated. The plasma glucagon levels were significantly lower over the 240-min postprandial period with GLP-1 treatment. The beneficial effects of GLP-1 on plasma glucose, insulin and glucagon concentrations were fully maintained for the 3-week treatment period. 4. We have demonstrated a significant improvement in postprandial glycaemic control with subcutaneous GLP-1 treatment. GLP-1 improves glycaemic control partially by restoring the first-phase insulin response and suppressing glucagon and is a potential treatment for Type 2 diabetes. PMID- 9730853 TI - Glycoxidation, and protein and DNA oxidation in patients with diabetes mellitus. AB - 1. The role of oxidative stress in the pathogenesis of the diabetic state is being investigated extensively. Although oxidative stress has been reported in terms of glycoxidation, protein oxidation and DNA oxidation in diabetes mellitus, oxidation parameters have not been determined in parallel on the same study population.2. We studied 24 patients with diabetes mellitus (14 patients with Type I diabetes with a mean age of 62.3+/-6.3 years and 10 patients with Type II diabetes aged 67.3+/-5.9 years) and compared them with age-matched non-diabetic controls. Urinary o-tyrosine, 8-hydroxy-2'-deoxyguanosine and pentosidine measurements by HPLC were made on two occasions (t1 and t2).3.A clear statistical difference was found between diabetic patients and controls at t1 or t2 for 8 hydroxy-2'-deoxyguanosine and pentosidine, but not for o-tyrosine. No significant correlations were found between clinical and other laboratory parameters except high-density lipoprotein and uric acid. We revealed significantly increased glycoxidation and DNA oxidation in patients with Type I and Type II diabetes, but protein oxidation was not different from controls.4. The finding of increased glycoxidation reflects increased oxidation of the carbohydrate moiety, whereas the increased levels of oxidized DNA may also be interpreted as due to increased DNA repair. The increased 8-hydroxy-2'-deoxyguanosine does not indicate the generation of an individual active oxygen species, but DNA could have been oxidized simply by alkenals from lipid peroxidation, as e.g. malondialdehyde. As no difference in protein oxidation (i.e. o-tyrosine) between diabetics and controls could be revealed, the oxidation of DNA by hydroxyl radical attack is unlikely, as o-tyrosine was proposed as a marker for hydroxyl radical attack. Therefore, the message is that increased glycoxidation can be confirmed, protein oxidation does not appear to take place and increased DNA oxidation is still not proven, as increased 8-hydroxy-2'-deoxyguanosine may simply reflect repair. PMID- 9730854 TI - Absence of glutamine isotopic steady state: implications for the assessment of whole-body glutamine production rate. AB - 1. During infusion of [5-15N]glutamine in patients with gastrointestinal cancer we unexpectedly observed a gradual decrease in time of the appearance rate (Ra) of glutamine in plasma. Here we investigate whether the failure to achieve a plateau isotopic enrichment in plasma is, among other factors, due to incomplete equilibration of the glutamine tracer with the large intramuscular free glutamine pool.2. Plasma and intramuscular glutamine enrichment were measured during 6-11 h infusions of L-[5-15N]glutamine and L-[1-13C]glutamine in post-absorptive patients admitted to hospital for elective abdominal surgery. L-[1-13C]Leucine and L-[ring-2H5]phenylalanine were infused to measure the proportion of glutamine appearing in plasma directly due to its release from protein.3. The glutamine tracer entered muscle, but the rise in intramuscular glutamine enrichment was small, presumably as a result of the enormous size of the intramuscular glutamine pool and the limited speed of entry of glutamine into muscle. In each patient the intramuscular glutamine enrichment was lower than that in plasma (P<0.001), and both increased with tracer infusion time (P<0.001), indicating incomplete equilibration of the glutamine tracer.4.A comparison of the results obtained by the two glutamine tracers indicated that recycling of the nitrogen label contributed to about 15% of the decrease in Ra.5. There was a gradual reduction in the glutamine release from proteolysis, which contributed to 16-21% of the decline in Ra.6. We conclude that slow equilibration of the glutamine tracer with the large muscle glutamine pool significantly contributes to the absence of isotopic steady state. Consequently, the appearance rate of glutamine in plasma measured during short tracer infusion periods (hours) considerably overestimates the whole-body glutamine flux. PMID- 9730855 TI - Effect of interleukin-4 on pro-inflammatory cytokine production and the acute phase response in healthy individuals and in patients with cancer or multiple organ failure. AB - 1. This study investigates whether previously documented effects of interleukin-4 in down-regulating pro-inflammatory cytokine production by peripheral blood mononuclear cells (PBMCs) from healthy individuals would be reproducible in PBMCs isolated from patients with multiple organ failure (acute disease model) and gastrointestinal cancer (chronic disease model). The effects of interleukin-4 on the ability of PBMC supernatants to elicit an acute phase protein response from isolated human hepatocytes were also studied. 2. Incubation of PBMCs with interleukin-4 significantly reduced both spontaneous and lipopolysaccharide induced production of tumour necrosis factor and lipopolysaccharide-induced interleukin-6 production, demonstrating that the PBMCs from patients with acute and chronic disease are not refractory to the effects of interleukin-4. The effects of interleukin-4 on the ability of PBMCs from the groups studied to elicit an acute phase response were complex and varied both between patient groups and individual acute phase proteins. Overall, interleukin-4 reduced the potential of PBMCs to stimulate production of the positive acute phase proteins C reactive protein, alpha1-antichymotrypsin and alpha1-acid glycoprotein.3. This work emphasizes the pleiotropic nature of cytokines and the complex regulatory mechanisms which exist. The study illustrates the difficulties in devising in vivo intervention strategies using cytokines such as interleukin-4. PMID- 9730856 TI - Nitric oxide synthase activity is increased in relation to the severity of liver dysfunction. AB - 1. Nitric oxide is a potent vasodilator which plays a major role in the control of blood pressure. The hyperdynamic circulation of cirrhosis has been linked to nitric oxide.2. We measured neutrophil nitric oxide synthase activity in relation to the level of hepatic dysfunction in patients with liver disease of varying aetiology and severity.3. Neutrophils were isolated from 21 patients (7 Child Pugh score A, 6 grade B and 8 grade C) aged 28-76 (median 49) years. Nitric oxide synthase activity was measured using the conversion of oxyhaemoglobin to methaemoglobin by nitric oxide and expressed in terms of cell protein. Blood pressure and biochemical indices were recorded. Data were assessed using Kruskal Wallis one-way analysis of variance, Mann-Whitney U-test or Pearson correlation as appropriate.4.Systolic, mean arterial and diastolic blood pressures decreased with increasing hepatic damage (P=0.031, P=0.01 and P=0. 038 respectively). Nitric oxide synthase activity increased with the degree of liver dysfunction (P=0.033) and was highest in patients with Child-Pugh score C. Systolic blood pressure correlated with nitric oxide synthase activity in patients with Child Pugh score C (P=0.029).5. Our results show that nitric oxide synthase activity increases with increasing Child-Pugh score and is associated with the development of systemic hypotension. These data may support the involvement of nitric oxide in the haemodynamic disturbances seen in liver disease. PMID- 9730857 TI - Beneficial effect of vitamin E administration on nitric oxide function in subjects with hypercholesterolaemia. AB - 1. Vitamin E administration improves endothelial function in hypercholesterolaemic animals but, generally, has not been found to do so in man. The aim of this study was to determine whether vitamin E administration improves basal or stimulated function of the nitric oxide (NO) dilator system in patients with hypercholesterolaemia. 2. Seven subjects aged 47+/-3 (+/-S.E.M.) years with moderately elevated serum cholesterol concentrations (6.0+/-0.1 mmol/l) were given 4 weeks of placebo therapy followed by 500 i.u. of vitamin E twice daily for 4 weeks. Endothelium-dependent and -independent vasodilatation were assessed by intrabrachial infusion of acetylcholine and sodium nitroprusside, and forearm blood flow was measured by strain-gauge plethysmography. Basal NO function was assessed by infusion of NG-monomethyl-L-arginine. 3. Plasma alpha-tocopherol concentration was enhanced after administration of vitamin E (34.6+/-1.8 to 86.9+/-9.6 micromol/l; P<0.001). In addition, vitamin E administration significantly increased acetylcholine-mediated vasodilatation whether the results were expressed in terms of changes in absolute forearm blood flow (P<0. 01), forearm vascular resistance (P<0.05) or forearm blood flow ratios (P<0.001). Similarly, absolute forearm blood flow (P<0.05), forearm vascular resistance (P<0.01) and forearm blood flow ratio (P<0.01) responses to NG-monomethyl-L arginine were augmented by vitamin E therapy. Sodium nitroprusside responses were unaltered. 4. These results indicate that 4 weeks therapy with 1000 i.u. of vitamin E daily improves basal and stimulated NO-related endothelial function in subjects with hypercholesterolaemia. PMID- 9730858 TI - Protective agents used as additives in University of Wisconsin solution to promote protection against ischaemia-reperfusion injury in rat lung. AB - 1. An intervention to reduce ischaemia-reperfusion lung injury will be an important advance in transplant medicine. Although the mechanisms associated with producing ischaemia-reperfusion endothelial injury have not been completely elucidated, many of the injury mediators have been studied in detail. While no single pharmacological therapy is likely to be totally effective in eliminating this complex injury, we have developed a mixture of agents that are known to block pathways involved in producing ischaemia-reperfusion-associated lung vascular injury.2. The present study modified University of Wisconsin solution (UW) by adding one of the protective agents prostaglandin E1 (PGE1), dexamethasone (Dex) or dibutyryl cAMP (Bt2-cAMP), or a combination of these, to the perfusate of rat lungs exposed to 4 h of cold ischaemia followed by 1 h of reperfusion. Nine modified UW solutions were studied: (1) UW+Dex, (2) UW+PGE1, (3) UW+Bt2-cAMP, (4) UW+Dexx3, (5) UW+PGE1x3, (6) UW+Bt2-cAMPx3, (7) UW+Dex+PGE1, (8) UW+Dex+Bt2-cAMP, (9) UW+PGE1+Bt2-cAMP. These solutions were utilized in individual experiments to assess haemodynamic changes, lung weight gain, the capillary filtration coefficient (Kfc) and pathology in all lungs.3. The results indicate that lung weight gain and Kfc values were significantly lower than with UW alone in groups 1, 2 and 3, which contained only one additional protective agent. In groups 4, 5 and 6, which contain three times the concentration of each protective agent, both Kfc and lung weight gain were similar to those measured in groups 1, 2 and 3, i.e. lungs were protected but the protection was not dose dependent. In groups 7, 8 and 9, which contained two protective agents, lung weight gain and Kfc were greatly reduced compared with UW alone. Histopathological studies showed similar decreases in the injury profiles of lungs.4. Although UW contains several antioxidant protective agents such as allopurinol and glutathione, it did not provide effective protection in our ischaemia-reperfusion lung injury model. UW modified with an additive of PGE1, Dex or Bt2-cAMP attenuated ischaemia-reperfusion injury. Furthermore, UW containing two of these protective agents augmented the protection. Among the modified solutions, it appears that UW+PGE1+Bt2-cAMP protects the lungs to a greater extent than all other solutions used in our study. We suggest that preservation solutions containing PGE1-Bt2-cAMP will provide additional protective effects to organs stored for transplantation. PMID- 9730859 TI - Regulation of cAMP in a lymphocyte preparation isolated from peripheral venous blood in human subjects: the significance of residual thrombocytes, noradrenaline and prostaglandins. AB - 1. The aim of the study was to elucidate the mechanism of the previously reported close correlation observed between noradrenaline and cAMP in a lymphocyte preparation (LP) isolated from peripheral venous blood in healthy subjects. A close correlation was also obtained in the present study between lymphocyte noradrenaline and adrenaline and cAMP both in the basal state and after stimulation with isoproterenol (P<0.05 to 0.007).2. Although 99% of the thrombocytes were removed from the LP during the washing procedure, LP contained approximately one thrombocyte per lymphocyte. The noradrenaline concentration in LP which could be ascribed to residual thrombocytes, calculated from the average noradrenaline concentration in thrombocytes and the number of thrombocytes in LP, correlated closely to noradrenaline in LP (P<0.007).3. To test the hypothesis that noradrenaline in LP was primarily derived from plasma, we studied three patients with phaeochromocytoma, who had high levels of noradrenaline and adrenaline both in plasma and in LP. 4. Further studies showed that the addition of thrombocytes to LP increased cAMP. The response was inhibited by indomethacin, whereas the addition of cimetidine and propranolol had no effect on basal or thrombocyte-stimulated cAMP.5.We conclude that noradrenaline in LP is a marker of the number of residual thrombocytes. The addition of thrombocytes to LP increased cAMP in lymphocytes. This response was not mediated by catecholamines but possibly by prostaglandins. PMID- 9730860 TI - Stimulation of apoptosis by sulindac and piroxicam. AB - 1.Sulindac, cis-5-fluoro-2-methyl-1-(p-methylsulphinylbenzylidene)indene-3-ace tic acid, inhibits growth of colon polyps and cancers. This effect has been attributed to inhibition of prostaglandin synthesis but more recent observations indicate that, in vitro, cells that do not have cyclo-oxygenase nor RNA for synthesis of such enzymes are affected by sulindac. Therefore the presumptive effect is probably not correct.2.It has also been found that sulindac stimulates apoptosis. It is herein postulated that in tumour cells such effects may be due to interaction of the anionic form of the drug with protons in the intermembrane space of mitochondria to disrupt the potential across the inner mitochondrial membrane and thereby initiate apoptosis. Normal cells are not affected. PMID- 9730861 TI - Airway epithelial integrins: why so many? PMID- 9730862 TI - Antibacterial peptides in bronchoalveolar lavage fluid. AB - Defensins and other antimicrobial peptides act in the innate host defense of epithelial surfaces. Human beta defensin 1 (hBD-1) has recently been shown to be expressed in airway epithelial cells and so has been implicated as a primary component of antibacterial activity in human lung. We attempted to purify these molecules from bronchoalveolar lavage fluid (BALF). Extraction of BALF on SepPak C-18 cartridges, followed by continuous acid-urea polyacrylamide gel electrophoresis and reverse-phase high-performance liquid chromatography yielded one fraction with antibacterial activity associated with factors of < 6.5 kD. N terminal amino acid sequencing identified these peptides as human neutrophil defensins (HD) 1 through 3. No hBD-1 was detected. Together with lysozyme, it appears that HD-1 through -3 are the most prominent antimicrobial factors in BALF. The contribution of epithelial defensins such as hBD-1 to antibacterial defense of human airway in vivo remains to be elucidated. PMID- 9730863 TI - Ozone alters the distribution of beta1 integrins in cultured primate bronchial epithelial cells. AB - The effects of 0.5 ppm ozone exposure for 6 h on the synthesis and distribution of beta1 integrins were examined in bronchial epithelial cells cultured at an air cell interface. Ozone exposure damaged cilia and caused significant cell loss. Immunocytochemical localization and quantification of the beta1 subunit in the remaining attached cells using scanning laser cytometry demonstrated time dependent changes in beta1 distribution in response to ozone. Although no changes were detected immediately after exposure, beta1 immunoreactivity increased 23 +/- 5% and 66 +/- 6% at 6 and 24 h, respectively. The increased immunostaining was localized at the apical surfaces and, to a lesser extent, at cell-cell contacts of cultured cells. Furthermore, integrin redistribution was not due to increased messenger RNA (mRNA) levels and protein synthesis because levels of beta1 mRNA and newly synthesized beta1 protein did not change after ozone exposure. However, immunoprecipitation analysis of beta1 integrins in lysates from equal numbers of cells showed that ozone-exposed cells contained 90 +/- 15% more total beta1 subunit at 24 h after exposure. In addition, our results demonstrated the presence of the alpha5beta1 integrin complex in bronchial epithelial cells and that the detergent-soluble amount of its associated beta1 subunit increased 60 +/ 10% in lysates of ozone-exposed cells. In conclusion, ozone altered cellular distribution of beta1 integrins in the remaining attached cells subsequent to cell injury and loss. The changes in beta1 distribution might be due to increased detergent extractibility of beta1 integrins rather than a real increase in the synthesis of beta1 integrins. PMID- 9730864 TI - Copper-dependent inflammation and nuclear factor-kappaB activation by particulate air pollution. AB - Particulate air pollution causes increased cardiopulmonary morbidity and mortality, but the chemical determinants responsible for its biologic effects are not understood. We studied the effect of total suspended particulates collected in Provo, Utah, an area where an increase in respiratory symptoms in relation to levels of particulate pollution has been well documented. Provo particulates caused cytokine-induced neutrophil chemoattractant-dependent inflammation of rat lungs. Provo particulates stimulated interleukin-6 (IL-6) and IL-8 production, increased IL-8 messenger RNA (mRNA) and enhanced expression of intercellular adhesion molecule-1 (ICAM-1) in cultured BEAS-2B cells, and stimulated IL-8 secretion in primary cultures of human bronchial epithelium. Cytokine secretion was preceded by activation of the transcription factor nuclear factor-kappaB (NF kappaB) and was reduced by treatment of cultures with superoxide dismutase, deferoxamine, or N-acetylcysteine. These biologic effects were replicated by culturing BEAS cells with quantities of Cu2+ found in Provo extract. IL-8 secretion by BEAS cells could be modified by addition of normal constituents of airway lining fluid to the culture medium. Mucin significantly reduced IL-8 secretion, and ceruloplasmin significantly increased IL-8 secretion and activation of NF-kappaB. These findings suggest that copper ions may cause some of the biologic effects of inhaled particulate air pollution in the Provo region of the United States, and may provide an explanation for the sensitivity of asthmatic individuals to Provo particulates that has been observed in epidemiologic studies. PMID- 9730865 TI - Immune cells in a mouse airway model of obliterative bronchiolitis. AB - Obliterative bronchiolitis (OB), a form of chronic lung rejection, affects 50% of all lung-transplant recipients and is a major cause of morbidity and mortality. We used the mouse tracheal allograft model of OB to quantitate inflammatory cells during disease progression to evaluate the pathogenesis of this disorder. Tracheas of BALB/c mice were implanted into C57BL/6, severe combined immunodeficiency (SCID), and BALB/c mice. Cyclosporin was administered at 25 mg/kg/d. Grafts were harvested at 2, 6, 10, and 15 wk, and analyzed immunohistochemically. Tracheal allografts developed epithelial injury and cellular infiltrates at 2 wk, epithelial denudation and complete luminal obliteration at 6 wk, and dense collagenous scarring by 15 wk. SCID allografts and isografts demonstrated intact epithelium throughout, although a mononuclear infiltrate was initially present at 2 wk in the SCID allografts. Immunohistochemical staining, using antibodies to mouse CD4(+) (T-helper lymphocyte), CD8(+) (T-cytotoxic/suppressor lymphocyte), and B lymphocytes, macrophages, and myofibroblasts, revealed large numbers of macrophages and CD4(+) and CD8(+) lymphocytes in allografts at 2 wk, compared with isografts. The allograft CD4(+)/CD8(+) ratio was 0.75 at 2 wk. Allografts demonstrated macrophage, myofibroblast, and CD4(+) predominance at 6 and 10 wk (CD4(+)/CD8(+) = 2/1), but by 15 wk had minimal cellularity and were densely scarred. SCID allografts demonstrated a macrophage-predominant infiltrate at 2 wk, with minimal cellularity at later time points. These results indicate that: (1) OB is predominantly an immunologic airway injury; and (2) CD4(+) and CD8(+) lymphocytes and macrophages play an important role in the evolution of airway inflammation and fibrosis. Additionally, this model suggests that chronic airway fibrosis follows a period of intense airway-directed, cell-mediated rejection. PMID- 9730866 TI - Relationship of inhaled ozone concentration to acute tracheobronchial epithelial injury, site-specific ozone dose, and glutathione depletion in rhesus monkeys. AB - Acute pulmonary epithelial injury produced by short-term exposure to ozone varies by site within the tracheobronchial tree. To test whether this variability is related to the local dose of ozone at the tissue site or to local concentrations of glutathione, we exposed adult male rhesus monkeys for 2 h to filtered air or to 0.4 or 1.0 ppm ozone generated from 18O2. Following exposure, lungs were split into lobes and specimens were selected by microdissection so that measurements could be made on airway tissue of similar branching history, including trachea, proximal (generation one or two) and distal (generation six or seven) intrapulmonary bronchi, and proximal respiratory bronchioles. One half of the lung was lavaged for analysis of extracellular components. In monkeys exposed to filtered air, the concentration of reduced glutathione (GSH) varied throughout the airway tree, with the proximal intrapulmonary bronchus having the lowest concentration and the parenchyma having the highest concentration. Exposure to 1.0 ppm ozone significantly reduced GSH only in the respiratory bronchiole, whereas exposure to 0.4 ppm increased GSH only in the proximal intrapulmonary bronchus. Local ozone dose (measured as excess 18O) varied by as much as a factor of three in different airways of monkeys exposed to 1.0 ppm, with respiratory bronchioles having the highest concentration and the parenchyma the lowest concentration. In monkeys exposed to 0.4 ppm, the ozone dose was 60% to 70% less than in the same site in monkeys exposed to 1.0 ppm. Epithelial disruption was present to some degree in all airway sites, but not in the parenchyma, in animals exposed to 1.0 ppm ozone. The mass of mucous and ciliated cells decreased in all airways, and necrotic and inflammatory cells increased. At 0.4 ppm, epithelial injury was minimal, except in the respiratory bronchiole, where cell loss and necrosis occurred, and was 50% that found in monkeys exposed to 1.0 ppm ozone. We conclude that there is a close association between site-specific O3 dose, the degree of epithelial injury, and glutathione depletion at local sites in the tracheobronchial tree. PMID- 9730867 TI - Beta-adrenoceptor-mediated inhibition of IFN-gamma, IL-3, and GM-CSF mRNA accumulation in activated human T lymphocytes is solely mediated by the beta2 adrenoceptor subtype. AB - Cytokine gene expression in T lymphocytes is a strictly regulated process, involving both stimulatory and inhibitory signals. beta-Adrenoceptor (betaAR) agonists are widely used in the treatment of asthma and are able to induce an inhibitory signal on immunological responses after binding to their specific receptors. In this study, the characterization of betaAR subtype(s) (beta1, beta2, and beta3) involved in the regulation of interleukin (IL)-3, IL-4, granulocyte-macrophage colony-stimulating factor (GM-CSF), and interferon-gamma (IFN-gamma) mRNA accumulation was studied by using various betaAR agonists and antagonists. Concanavalin A (Con A)-induced IFN-gamma, GM-CSF, and IL-3 mRNAs are dose-dependently inhibited by the nonselective betaAR agonist isoproterenol and by the selective beta2AR agonist fenoterol. IL-4 mRNA accumulation was not susceptible to betaAR stimulation. The observed inhibition on IFN-gamma, GM-CSF, and IL-3 mRNA was blocked by the selective beta2AR antagonist ICI 118,551 (10(-6) M) and by timolol (10(-6) M), a nonselective antagonist. The selective beta1AR antagonist atenolol (0.3 x 10(-6) M) did not have any effect. Secretion of GM-CSF protein in the presence of increasing concentrations of isoproterenol followed a similar pattern as observed for GM-CSF mRNA. In addition, the betaAR-mediated inhibition of IFN-gamma, GM-CSF, and IL-3 mRNA accumulation and GM-CSF protein secretion were related to the accumulation of intracellular cyclic adenosine monophosphate (cAMP) levels. Although beta3AR mRNA was detectable in Con A activated T lymphocytes, we could not demonstrate a functional activity in the regulation of cytokine expression: the beta3AR agonist BRL 37344 had no effect on the accumulation of the studied cytokine mRNAs, and did not significantly affect cellular cAMP levels. These data demonstrate that beta-agonist-induced inhibition of IFN-gamma, GM-CSF, and IL-3 mRNA accumulation is solely mediated by beta2 adrenoceptors. PMID- 9730868 TI - A CD18/ICAM-1-dependent pathway mediates eosinophil adhesion to human bronchial epithelial cells. AB - Eosinophil adhesion to airway epithelium is believed to facilitate eosinophil accumulation and retention in asthmatic airways. Monoclonal antibodies (mAb) against intercellular adhesion molecule-1 (ICAM-1) and its CD18 leukocyte integrin ligands have been shown to inhibit airway eosinophilia in animal models of asthma, although the role of this pathway in eosinophil-epithelial adhesion is not fully understood. To investigate the role in vitro of CD18 and ICAM-1, we measured adhesion of fluorescently labeled human eosinophils to normal human bronchial epithelial cell (NHBEC) monolayers pretreated for 24 h with culture medium (low constitutive ICAM-1) or tumor necrosis factor-alpha (TNF-alpha; 1 ng/ml) and interferon-gamma (IFN-gamma) (10 ng/ml; increased ICAM-1). Stimulation of eosinophils with C5a (10(-7) M) increased adhesion measured at 30 min to unactivated NHBEC from 11.4 +/- 0.7 to 15.5 +/- 0.4% (n = 4), and this increase was CD18/ICAM-1-independent, whereas phorbolmyristate acetate (PMA) (10(-8) M) induced adhesion (20.7 +/- 1.7%) was abolished by anti-CD18 and reduced by anti ICAM-1. In contrast, C5a- and PMA-induced adhesion to TNF-alpha/IFN-gamma activated NHBEC (increased from 11.1 +/- 1.3% to 21.9 +/- 1.0% and 27.6 +/- 1.9%, respectively) was CD18- and ICAM-1-dependent. Eotaxin, but not regulated on activation normal T cells expressed and secreted, macrophage inflammatory protein 1, formyl methionyl leucyl phenylalanine, leukotriene B4 or platelet-activating factor, also induced CD18/ICAM-1-dependent adhesion to activated NHBEC. In the absence of added chemoattractants, eosinophil adhesion to NHBEC increased with time and, at 120 min, was significantly greater (P < 0.01) to activated NHBEC (37.3 +/- 2.4%, n = 5) than to unactivated monolayers (24.3 +/- 1.9%); mAb against CD18 or ICAM-1 abolished increased, but not basal, adhesion. These results suggest that CD18/ICAM-1 mediated eosinophil adhesion to activated NHBEC but that adhesion to resting NHBEC was largely independent of this pathway. PMID- 9730869 TI - Influence of mechanical stretch on thrombin regulation by fetal mixed lung cells. AB - Respiratory distress syndrome (RDS) is characterized by intrapulmonary fibrin deposition, which can adversely affect surfactant function, and stimulate fibroblast proliferation, which may contribute to the development of bronchopulmonary dysplasia (BPD). We speculated that the premature lung may have impaired regulation of thrombin, thus making preterm infants susceptible to fibrin formation within the lung. Therefore, we studied the effect of stretch, which simulates fetal breathing movements (FBMs), on the generation and inhibition of a key hemostatic enzyme-thrombin-by rat fetal mixed lung cells (FMLCs). Our results showed that stretch induced glycosaminoglycan production with increased antithrombin activity due to an increase in the concentration of active chondroitin sulfate. Stretch downregulated secretion of tissue factor procoagulant activity, which may lead to decreased thrombin generation on the surface of FMLCs. Overall, stretch enhanced the local control of thrombin by FMLCs. These results suggest that premature infants, who will have experienced less FBM, may have impaired thrombin regulation. Impaired thrombin regulation likely contributes to increased fibrin deposition and, potentially, the development of BPD. PMID- 9730870 TI - The effects of hyperoxic injury and antioxidant vitamins on death and proliferation of human small airway epithelial cells. AB - Previously it was reported that hyperoxia induced death of the human lung adenocarcinoma cell line (A549 cells) by necrosis, not by apoptosis. This study examined proliferation and death of untransformed human small airway epithelial (SAE) cells in normoxia or hyperoxia in comparison with A549 cells. We tested the hypothesis that SAE cells respond differently to hyperoxic injury than do A549 cells. We measured total cell number and viability, thymidine incorporation (SAE cells only), lactate dehydrogenase (LDH) release, and apoptotic changes as markers for cell proliferation and death. Protective effects of antioxidant vitamins also were examined in SAE cells. In normoxia, subconfluent SAE cells had less apoptosis and fewer detached cells, but higher thymidine incorporation than did near-confluent cells. Hyperoxia suppressed thymidine incorporation and augmented apoptosis in both subconfluent and near-confluent SAE cells. Hyperoxia decreased the total cell number only in subconfluence, whereas SAE cell viability declined with hyperoxia in near confluence, but not in subconfluence. For SAE cells, necrosis assessed by LDH release was minimal in all conditions and was not augmented by hyperoxia in SAE cells. In contrast, normoxic A549 cells proliferated more rapidly than did SAE cells with a large number of cells detached during the culture. A549 cells underwent necrotic cell death under confluent or in hyperoxic conditions, but had much less apoptotic cell death. In SAE cells, vitamin E partially prevented the decline of thymidine incorporation with hyperoxia in subconfluence and protected against apoptotic changes with hyperoxia in both subconfluent and near-confluent conditions. Vitamin C prevented apoptosis with hyperoxia only in near-confluent SAE cells. Thus, SAE cells maintained balanced apoptosis and cell proliferation that were altered by cell density and hyperoxia and demonstrated very little necrosis with hyperoxia. Although A549 cells underwent cell death mainly by necrosis, they also were influenced by cell density and hyperoxia. Cell density also determined specific antioxidant vitamin protection in SAE cells. PMID- 9730871 TI - Calcium signaling in airway smooth muscle cells is altered by in vitro exposure to the aldehyde acrolein. AB - We have previously observed that acrolein administered ex vivo to isolated airways alters the subsequent airway responsiveness. To examine the cellular mechanisms involved in this alteration, we have studied the effect of acrolein exposure on calcium signaling in myocytes freshly isolated from rat trachea. We have also studied the effect of acrolein exposure on isometric contraction of rat epithelium-free tracheal rings. Tissues were exposed to a variety of acrolein concentrations from 0.1 to 1 microM and durations from 5 to 15 min. In isolated cells, exposure to acrolein did not modify the resting cytosolic Ca2+ concentration ([Ca2+]i) whatever the concentration or duration of exposure, but altered the pattern of the Ca2+ response to acetylcholine (ACh). ACh typically induces an initial [Ca2+]i rise followed by peaks of decreasing amplitude (oscillations). Exposure to a fixed concentration of acrolein (0.2 microM) for 5 and 10 min significantly enhanced the amplitude of the initial [Ca2+]i rise in response to a low concentration of ACh (0.1 microM) by 50.8 and 77%, respectively. Similarly, exposure for a fixed duration of 10 min significantly enhanced the amplitude of the initial [Ca2+]i rise by 49.4% at an acrolein concentration of 0.3 microM. When cells were stimulated with a high ACh concentration (10 microM), the value of the first [Ca2+]i peak was not changed by acrolein exposure; but the frequency at which subsequent peaks occurred was significantly increased by 44.4% after 10 min of exposure to a fixed concentration of 0.2 microM and by 36.3% following an exposure for a fixed duration of 10 min at the concentration of 0.3 microM. In contrast, acrolein, whatever the concentration, had no effect on the caffeine-induced [Ca2+]i response. In rat epithelium-free tracheal rings, acrolein increased the response to muscarinic stimulation, with a maximal effect observed for an exposure to 0.3 microM for 10 min. The effect of acrolein on the [Ca2+]i response of isolated myocytes occurred over a range of doses similar to that on the contractile response of rings, suggesting that the effect of this pollutant on calcium signaling may account, at least partially, for acrolein-induced airway hyperresponsiveness. PMID- 9730872 TI - Tenascin and fibronectin expression in human mesothelial cells and pleural mesothelioma cell-line cells. AB - Fibronectin (Fn) and tenascin (Tn) are two major extracellular matrix (ECM) glycoproteins that may have important roles both in fibrotic lung diseases and in lung tumors. The significance of Fn and Tn in human pleural mesothelial cells and pleural diseases is unclear. Transformed human pleural mesothelial cells (Met5A), primary cultures of mesothelial cells, and cultured mesothelioma cell lines were investigated for Fn and Tn immunoreactivity. Mesothelial cells were exposed for 48 to 96 h to transforming growth factor-beta (TGF-beta), tumor necrosis factor alpha (TNF-alpha), amosite asbestos fibers, or oxidants (H2O2 and menadione, a compound that auto-oxidizes to produce superoxide). Immunofluorescence and Western blotting with monoclonal anti-Fn and anti-Tn antibodies, and Northern blotting with a complementary DNA (cDNA) probe for Tn showed that mesothelial cells are capable of producing Fn and Tn. The mRNA level and immunoreactivity of Tn was enhanced by TGF-beta and TNF-alpha, whereas Fn was intensified only by TGF beta. A wide range of amosite, H2O2, or menadione concentrations had no clear effect on Fn or Tn reactivity. Fn and Tn were present at low or undetectable concentrations in five of six mesothelioma cell lines, whereas the organization of Fn immunoreactivity in these cell lines was variable. Furthermore, results obtained with the tumor tissue of these same mesothelioma patients suggested that Fn and Tn expressions do not necessarily parallel either each other or results obtained with the cultured cells. PMID- 9730873 TI - Effects of LTD4 on human airway smooth muscle cell proliferation, matrix expression, and contraction In vitro: differential sensitivity to cysteinyl leukotriene receptor antagonists. AB - The cysteinyl leukotrienes (CysLTs) mimic many of the features of asthma and are implicated in its pathophysiology. Little, however, is known about the effects of the CysLTs on airways remodeling. In this study the effects of leukotriene D4 (LTD4) on human airway smooth muscle (HASM) cell proliferation and expression of extracellular matrix proteins were investigated. LTD4 (0.1-10 microM) alone had no effect on DNA synthesis in HASM. LTD4, however, markedly augmented proliferation induced by the mitogen, epidermal growth factor (EGF, 1 ng/ml). The potentiating effect of LTD4 (1 microM) on EGF-induced DNA synthesis was abolished by pranlukast (1 microM) or pobilukast (30 microM), but unaffected by zafirlukast (1 microM). In contrast, pranlukast (pKB = 6.9), pobilukast (pKB = 7.0), and zafirlukast (pKB = 6.5) had equivalent potencies for inhibition of LTD4-induced contraction in human bronchus. LTD4 (0.1 or 10 microM) did not increase the total messenger RNA expression of the extracellular matrix proteins (pro-alpha[I] type I or alpha1[IV] type IV collagen), elastin, biglycan, decorin, and fibronectin, and did not influence tumor growth factor-beta (10 ng/ml)-induced effects on the expression of these proteins in HASM cells. These data indicate that LTD4 augments growth factor-induced HASM proliferation but does not alter the expression of various extracellular matrix components. The observed differences in sensitivity to the antagonists suggests that the former phenomenon may be mediated by a CysLT receptor distinct from that which mediates LTD4-induced HASM contraction. Collectively, these results provide preliminary evidence that CysLTs may play a role in airways remodeling in asthma. PMID- 9730874 TI - Altered alveolar macrophage function in calorie-restricted rats. AB - Alveolar macrophage functions associated with clearance of bacteria from the lung were assessed in male Fischer 344 rats maintained on a 25% calorie-restricted diet. Calorie-restricted and ad libitum-fed (control) rats were exposed to concentrations of ozone known to compromise phagocytic function of alveolar macrophages. Ozone suppressed alveolar macrophage phagocytosis of latex beads in vitro in ad libitum-fed rats, but not in calorie-restricted rats. In fact, caloric restriction enhanced phagocytic function in both control and ozone exposed animals. Ad libitum-fed rats exposed to ozone and challenged with Streptococcus zooepidemicus experienced a prolonged infection and influx of polymorphonuclear leukocytes (PMN), whereas calorie-restricted rats exposed to ozone cleared the bacteria in 24 h without an inflammatory response. Bacterial endotoxin-stimulated in vitro production of nitric oxide and tumor necrosis factor (TNF)-alpha as well as expression of TNF-alpha and interleukin-6 messenger RNAs were all lower in alveolar macrophages isolated from calorie-restricted rats. Together, the data suggest that caloric restriction enhances resistance to gram-positive bacteria, while lowering the production of proinflammatory mediators elicited by endotoxin, a component of gram-negative bacteria. Although increased bacterial resistance is considered beneficial, reduction in the lung's ability to induce inflammatory mediators can have both positive and pathophysiologic consequences. PMID- 9730875 TI - Production of endothelins by the ventilatory muscles in septic shock. AB - Circulating endothelin-1 (ET-1) concentration increases significantly in animal models of sepsis. The main mechanism responsible for this rise in ET-1 levels is believed to be upregulation of ET-1 synthesis in various organs, such as the lungs and heart. In this study we investigated whether ET-1 is synthesized in the ventilatory muscles and whether this synthesis is regulated in septic shock. Conscious rats were injected with Escherichia coli endotoxin (lipopolysaccharide [LPS]) and killed 6, 12, and 24 h later. A fourth group of rats was injected with normal saline and served as a control. The diaphragm was excised at the end of the experiment and quickly frozen. Diaphragmatic ET-1 level was measured with radioimmunoassay, and messenger RNA (mRNA) expression of ET-1 precursor prohormone (preproET-1), preproET-3, and endothelin-converting enzyme was measured with reverse transcription-polymerase chain reaction. LPS injection elicited an early (within 6 h) and prolonged rise in diaphragmatic ET-1 concentration. In addition, mRNA levels of preproET-1 and preproET-3 rose by about 4- and 3-fold within 6 to 12 h of LPS injection, whereas mRNA of endothelin converting enzyme increased by more than 10-fold and peaked within 24 h of LPS injection. Immunostaining with anti-ET-1 antibody revealed positive ET-1 staining in the endothelium and somatic muscle fibers of septic diaphragms. These results indicate that diaphragmatic muscle fibers synthesize significant amounts of ET-1 in septic shock and that the rise in ET-1 production is due to upregulation of ET precursors and the converting enzyme. PMID- 9730876 TI - Pharmacologic importance of the reversible fatty acid conjugation of budesonide studied in a rat cell line In vitro. AB - Functional implications of the recently described fatty acid conjugation of budesonide (BUD) (Tunek, A., K. Sjodin, and G. Hallstrom, Drug Metabol. Dispos. 1997;25:1311-1317; Miller-Larson, A., E. Hjertberg, H. Mattsson, M. Dahlback, A. Tunek, and R. Brattsand, Am. J. Respir. Crit. Care Med. 1997;155:A353 [Abstr.]) were studied in a rat cell line, Rat1, transfected with the activation protein-1 (AP-1)-controlled regulatory element (TRE) driving the reporter gene beta galactosidase. TRE is downregulated by glucocorticosteroids (GCS) through interaction with the AP-1 complex. BUD was compared to fluticasone propionate (FP), a potent glucocorticosteroid that does not form fatty acid conjugates. The kinetics and metabolism of the GCS were studied after incubation of either 3H-BUD or 3H-FP with transfected Rat1 cells. Up to 20% of added BUD was taken up into the cells over 24 h. The great majority of the intracellular radioactivity (80 90%) consisted of lipophilic BUD conjugates. After removing extracellular 3H-GCS, the outflow of radioactivity was studied. Only free BUD and not fatty acid conjugates was detected extracellularly, suggesting that hydrolysis of the conjugates was required to release BUD from the cell. During 165 min, less BUD (about 65% of totally incorporated) was released than FP (more than 90%). In the functional studies, FP was about six times more potent than BUD in downregulating TRE after 24 h continuous exposure. However, after a 6-h pulse of GCS, the effect of BUD persisted unchanged 18 h later, whereas FP had almost lost its efficacy (P < 0.05 between the drugs). In addition, the reversible conjugation process of BUD resulted in transferable GCS effects. Medium containing released BUD from previously loaded cells mediated nearly the same downregulatory effect after addition to naive cells as did continuous treatment. No such transferable effect was seen for FP. In conclusion, the reversible fatty acid conjugation of BUD resulted in prolonged cellular retention and anti-inflammatory activity after pulse exposure in this in vitro system. This fatty acid conjugation mechanism appears to add to the beneficial pharmacologic profile of BUD. PMID- 9730877 TI - Direct activation of K(Ca) channel in airway smooth muscle by nitric oxide: involvement of a nitrothiosylation mechanism? AB - Clinically, nitric oxide (NO*) is widely used as a pulmonary vaso- and bronchodilator agent. However, the precise molecular mechanisms by which NO. induces smooth muscle relaxation are not well established. It has been suggested that NO. relaxes airway smooth muscle (ASM) via a 3',5'-cyclic guanosine monophosphate (cGMP)-dependent pathway, and our previous work has shown that Ca2+ activated K+ (KCa) channels are susceptible to cGMP-dependent protein kinase (PKG)-dependent phosphorylation (A. Alioua, J. P. Huggins, and E. Rousseau. Am. J. Physiol. 1995;268:L1057-L1063). To assess whether KCa channels are also directly activated by NO. or one of its derivatives such as peroxynitrite, the activity of these channels was measured upon fusion of sarcolemmal vesicles derived from bovine tracheal smooth muscle cells into planar lipid bilayers (PLB). It was found that in the absence of adenosine triphosphate (ATP), cGMP, and cGMP-dependent protein kinase, NO* donors such as 1-propanamine-3-(2-hydroxy 2-nitroso-1-propylhydrazine) (PAPA NONOate) or 3-morpholinosydnonimine hydrochloride (SIN-1) in the presence of superoxide dismutase (SOD), added on either side of the bilayer, caused a concentration- dependent increase in the open probability (Po) of KCa channels without altering their unitary conductance. Release of NO*, which was measured by chemiluminescence analysis in parallel experiments, affected the gating behavior of KCa channels in the presence of SOD and ethyleneglycol-bis-(beta-aminoethyl ether)- N,N'-tetraacetic acid (EGTA) by reducing the mean closed times and increasing the number and duration of short open events. PAPA NONOate, a true NO. donor, had similar effects in the presence of ethylenediaminetetraacetic acid (EDTA), a heavy-metal chelator, and K-urate, a peroxynitrite scavenger. Addition of either 5 mM dithiothreitol (DTT) or 5 mM reduced glutathione (GSH), as well as 5 mM N-ethylmaleimide (NEM)-an alkylating agent-to the trans (intracellular) side of an experimental chamber slightly increased channel Po but prevented further channel activation by NO* donors. However, neither DTT nor GSH was able to reverse the effect of NO*. In contrast to SIN-1, DTT had no effect when added to the cis (extracellular) side of the chamber. This suggests that the effect of NO* is most likely due to a chemical modification (nitrothiosylation) of intracellular sulfhydryl group(s). Neither PAPA NONOate (NO*), nor SIN-1 had any effect on sarcolemmal Cl- channels reconstituted from the same membrane preparations. Pharmacomechanical measurements made on epithelium-denuded rat bronchus showed that 100 nM charybdotoxin decreased the sensitivity of bronchial smooth muscle to SIN-1 induced relaxations. Altogether, our data suggest that NO-induced bronchorelaxation occurs partly via a direct activation of KCa channels, possibly through a covalent interaction with the cytoplasmic side of their alpha subunit. PMID- 9730878 TI - Cadmium inhibits proteoglycan and procollagen production by cultured human lung fibroblasts. AB - Chronic inhalation of cadmium at the workplace or in cigarette smoke is associated with emphysema, a disease characterized by extensive disruption of lung connective tissue. We have previously shown that cadmium, at noncytotoxic doses, inhibits fibroblast procollagen production in vitro, with maximal inhibitory effects of 69 +/- 6% (P < 0.01) at 30 µM cadmium chloride (CdCl2). In this paper we show that at similar doses, cadmium also inhibits proteoglycan synthesis, with values reduced by between 36 +/- 4% (P < 0.01) and 42 +/- 6% (P < 0.01) for proteoglycans secreted into the culture media and associated with the cell layer, respectively. The greatest inhibition was obtained for the major matrix-associated proteoglycans, versican, decorin, and the large heparan sulfate proteoglycans, with synthesis values reduced by between 60 and 70%. Biglycan and other heparan sulfate proteoglycans were also affected, with synthesis values reduced by between 25 and 45%. In contrast, total protein synthesis was unaffected. Furthermore, effects of cadmium at the protein level were mirrored by reduction in messenger RNA levels for alpha1(I) procollagen, versican, and decorin. These data support the hypothesis that cadmium may play an important role in the pathogenesis of emphysema associated with chronic inhalation of cadmium fumes by inhibiting the production of connective tissue proteins. PMID- 9730879 TI - Combined nasal challenge with diesel exhaust particles and allergen induces In vivo IgE isotype switching. AB - In this study we undertook to provide evidence for local in vivo isotype switching to IgE following nasal challenges. Detection of deleted switch circular DNA (switch circles) by a novel nested polymerase chain reaction-based approach was employed as definitive molecular evidence of Ig isotype switching. Nasal challenge in humans with diesel exhaust particles (DEP) plus ragweed antigen has been shown to enhance local IgE production, stimulate local cytokine production, and markedly increase mucosal IgE antibody to ragweed. Four days after combined intranasal DEP plus ragweed challenge, we detected and characterized clones of deleted switch circular DNA (Sepsilon /Smu) representing switching from mu to epsilon from nasal lavage cells. No switch circular DNA was detected in nasal lavage cells following challenge with DEP alone nor with ragweed allergen alone. These results indicate that the combination of mucosal stimulation with DEP and ragweed allergen is capable of driving in vivo isotype switching to IgE in humans with ragweed allergy. These results are the first direct demonstration of in vivo IgE isotype switching in humans. PMID- 9730880 TI - Chemokines induced by infection of mononuclear phagocytes with mycobacteria and present in lung alveoli during active pulmonary tuberculosis. AB - The capacity of Mycobacterium tuberculosis (MTB) to induce production of chemokines with known chemotactic activity for monocytes and lymphocytes, the cellular building blocks of granulomas, was investigated. These chemokines included regulated upon activation, normal T cell expressed and secreted (RANTES), monocyte chemotactic protein-1 (MCP-1), and macrophage inflammatory protein-1alpha (MIP-1alpha). MTB stimulated production of MCP-1 and MIP-1alpha by blood monocytes (MN) and alveolar macrophages (AM). MTB infection of MN and AM stimulated release but not production of RANTES. AM produced or released significantly higher levels than MN of RANTES (by 2.1-fold), MCP-1 (by 6.9-fold), and MIP-1alpha (by 5. 5-fold) (P < 0.05 for each). This study also confirmed that MTB-infected AM produce the chemokine interleukin (IL)-8. MTB infection of AM resulted in increased steady-state expression of messenger RNA (mRNA) for MCP-1 and MIP-1alpha and minimal increased expression of RANTES mRNA. Both an avirulent (H37Ra) and a virulent (H37Rv) strain of MTB and purified protein derivative of H37Rv but not latex beads induced production of chemokines. Supernatants of MTB infected cells demonstrated chemotactic activity for both monocytes and lymphocytes partially inhibitable by neutralizing antibodies against the chemokines studied. Bronchoalveolar lavage fluid from patients with active pulmonary tuberculosis as compared with healthy control subjects contained increased levels of RANTES (by 8-fold), MCP-1 (by 2.7-fold), and IL-8 (by 8.9 fold) (P < 0.05), but not MIP-1alpha, as compared with healthy control subjects. Thus, multiple chemokines may be involved in recruitment of cells for granuloma formation in tuberculosis. PMID- 9730881 TI - Cytokines modulate expression of cell-membrane complement inhibitory proteins in human lung cancer cell lines. AB - Human lung cancers overexpress several cell-membrane complement inhibitory proteins (CIP). These complement inhibitory proteins are membrane cofactor protein (CD46), decay-accelerating factor (DAF; CD55), and CD59 (protectin). These cell-membrane proteins have a wide normal tissue distribution, are known to protect normal host cells from homologous complement-mediated lysis, and are thought to facilitate tumor escape from immunosurveillance. To study whether proinflammatory cytokines that are involved in cancer growth can modulate cell membrane CIP expression in lung cancer cells, we studied the effect of interleukin (IL)-1alpha, tumor necrosis factor (TNF)-alpha, and interferon (IFN) gamma on two human lung cancer cell lines. ChaGo K-1 and NCI-H596 cell lines, undifferentiated carcinoma and lung adenosquamous carcinoma, respectively, were stimulated with different cytokines, and the effects of incubation time and cytokine concentration on cell-membrane CIP expression were studied. Cell membrane CIP expression was evaluated using flow cytometry and cytokine effect was calculated as percent change in mean fluorescence intensity of each CIP molecule from its untreated control. We found that DAF was the lung cancer cell membrane CIP molecule that was the most responsive to cytokine stimulation. Maximal stimulatory effect was usually noted 72 h after a cytokine was introduced. In ChaGo K-1 and NCI-H596 lung cancer cell lines, IL-1alpha and TNF alpha increased DAF expression. IL-1alpha (100 U/ml/72 h) increased DAF expression up to a maximal mean of 45 and 48%, respectively, in comparison with untreated cells. TNF-alpha (1, 000 U/ml/72 h) increased DAF expression up to a mean of 131 and 46%, respectively. IFN-gamma (1 U/ml/72 h) increased DAF expression in NCI-H596 cells up to a mean of 100%, but had a slight inhibitory effect on DAF expression in ChaGo K-1 cells, decreasing expression by a mean of 17% in comparison with untreated cells. We conclude that cell-membrane DAF expression in the studied human lung cancer cell lines is modulated by IL-1alpha, TNF-alpha, and IFN-gamma, and speculate that cytokine-mediated modulation of cell membrane DAF in human lung cancer cells might affect lung cancer cell biology. PMID- 9730882 TI - Multidrug resistance protein 1 protects the oropharyngeal mucosal layer and the testicular tubules against drug-induced damage. AB - The multidrug resistance protein 1 (MRP1) gene encodes a transporter protein that helps to protect cells against xenobiotics. Elevated levels of MRP1 in tumor cells can result in active extrusion of a wide range of (anticancer) drugs with different cellular targets, a phenomenon called multidrug resistance (MDR). To explore the protective function of the mouse mrp1 protein during drug treatment, we investigated the toxicity caused by the anticancer drug etoposide-phosphate (ETOPOPHOS) in mice lacking the mrp1 gene (mrp1(-/-) mice). We show here that the lack of mrp1 protein results in increased etoposide-induced damage to the mucosa of the oropharyngeal cavity and to the seminiferous tubules of the testis. The high concentrations of mrp1 that we find in the basal layers of the oropharyngeal mucosa and in the basal membrane of the Sertoli cells in the testis apparently protect wild-type mice against this tissue damage. We also find drug-induced polyuria in mrp1(-/-) mice, which correlates with the presence of mrp1 protein in the urinary collecting tubules, the major site of kidney water reabsorption. Our results indicate that specific inhibitors of MRP1 used to reverse MDR, in combination with carcinostatic drugs transported by MRP1, might lead to drug induced mucositis, (temporary) infertility, and diabetes insipidus. PMID- 9730883 TI - Vaccination against chlamydial genital tract infection after immunization with dendritic cells pulsed ex vivo with nonviable Chlamydiae. AB - Chlamydia trachomatis, an obligate intracellular bacterial pathogen of mucosal surfaces, is a major cause of preventable blindness and sexually transmitted diseases for which vaccines are badly needed. Despite considerable effort, antichlamydial vaccines have proven to be elusive using conventional immunization strategies. We report the use of murine bone marrow-derived dendritic cells (DC) pulsed ex vivo with killed chlamydiae as a novel approach to vaccination against chlamydial infection. Our results show that DC efficiently phagocytose chlamydiae, secrete IL-12 p40, and present chlamydial antigen(s) to infection sensitized CD4(+) T cells. Mice immunized intravenously with chlamydial-pulsed DC produce protective immunity against chlamydial infection of the female genital tract equal to that obtained after infection with live organisms. Immunized mice shed approximately 3 logs fewer infectious chlamydiae and are protected from genital tract inflammatory and obstructive disease. Protective immunity is correlated with a chlamydial-specific Th1-biased response that closely mimics the immune response produced after chlamydial infection. Thus, ex vivo antigen-pulsed DC represent a powerful tool for the study of protective immunity to chlamydial mucosal infection and for the identification of chlamydial protective antigens through reconstitution experiments. Moreover, these findings might impact the design of vaccine strategies against other medically important sexually transmitted diseases for which vaccines are sought but which have proven difficult to develop. PMID- 9730884 TI - Syk tyrosine kinase and B cell antigen receptor (BCR) immunoglobulin-alpha subunit determine BCR-mediated major histocompatibility complex class II restricted antigen presentation. AB - Stimulation of CD4(+) helper T lymphocytes by antigen-presenting cells requires the degradation of exogenous antigens into antigenic peptides which associate with major histocompatibility complex (MHC) class II molecules in endosomal or lysosomal compartments. B lymphocytes mediate efficient antigen presentation first by capturing soluble antigens through clonally distributed antigen receptors (BCRs), composed of membrane immunoglobulin (Ig) associated with Ig alpha/Ig-beta heterodimers which, second, target antigens to MHC class II containing compartments. We report that antigen internalization and antigen targeting through the BCR or its Ig-alpha-associated subunit to newly synthesized class II lead to the presentation of a large spectrum of T cell epitopes, including some cryptic T cell epitopes. To further characterize the intracellular mechanisms of BCR-mediated antigen presentation, we used two complementary experimental approaches: mutational analysis of the Ig-alpha cytoplasmic tail, and overexpression in B cells of dominant negative syk mutants. Thus, we found that the syk tyrosine kinase, an effector of the BCR signal transduction pathway, is involved in the presentation of peptide- MHC class II complexes through antigen targeting by BCR subunits. PMID- 9730885 TI - Independent and opposing roles for Btk and lyn in B and myeloid signaling pathways. AB - Transphosphorylation by Src family kinases is required for the activation of Bruton's tyrosine kinase (Btk). Differences in the phenotypes of Btk-/- and lyn-/ mice suggest that these kinases may also have independent or opposing functions. B cell development and function were examined in Btk-/-lyn-/- mice to better understand the functional interaction of Btk and Lyn in vivo. The antigen independent phase of B lymphopoiesis was normal in Btk-/-lyn-/- mice. However, Btk-/-lyn-/- animals had a more severe immunodeficiency than Btk-/- mice. B cell numbers and response to T cell-dependent antigens were reduced. Btk and Lyn therefore play independent or partially redundant roles in the maintenance and function of peripheral B cells. Autoimmunity, hypersensitivity to B cell receptor (BCR) cross-linking, and splenomegaly caused by myeloerythroid hyperplasia were alleviated by Btk deficiency in lyn-/- mice. A transgene expressing Btk at approximately 25% of endogenous levels (Btklo) was crossed onto Btk-/- and Btk-/ lyn-/- backgrounds to demonstrate that Btk is limiting for BCR signaling in the presence but not in the absence of Lyn. These observations indicate that the net outcome of Lyn function in vivo is to inhibit Btk-dependent pathways in B and myeloid cells, and that Btklo mice are a useful sensitized system to identify regulatory components of Btk signaling pathways. PMID- 9730886 TI - Recombinant Mycobacterium bovis bacillus Calmette-Guerin secreting merozoite surface protein 1 (MSP1) induces protection against rodent malaria parasite infection depending on MSP1-stimulated interferon gamma and parasite-specific antibodies. AB - The merozoite surface protein 1 (MSP1) has emerged as a leading malaria vaccine candidate at the erythrocytic stage. Recombinant bacillus Calmette-Guerin (rBCG), which expressed a COOH-terminal 15-kD fragment of MSP1 of Plasmodium yoelii (MSP1 15) as a fusion protein with a secretory protein of Mycobacterium kansasii, was constructed. Immunization of mice with this rBCG induced a higher degree of protection against blood-stage parasite infection than with recombinant MSP1-15 in the RIBI adjuvant (RIBI ImmunoChem Research, Inc., Hamilton, MT) or incomplete Freund's adjuvant systems. We studied the mechanism of protection induced by MSP1 15, and found that interferon (IFN)-gamma had a major role in protection in all adjuvant systems we examined. Mice that produced low amounts of MSP1-15 stimulated IFN-gamma and could not control parasite infection. The antibody against MSP1-15 did not play a major role in protection in this system. After parasite infection, immunoglobulin G2a antibodies, which had been produced by IFN gamma stimulation, were induced and subsequently played an important role in eradicating parasites. Thus, both cellular and humoral immune responses were essential for protection from malaria disease. These data revealed that BCG is a powerful adjuvant to induce such a protective immune response against malaria parasites. PMID- 9730887 TI - Chemokine sequestration by viral chemoreceptors as a novel viral escape strategy: withdrawal of chemokines from the environment of cytomegalovirus-infected cells. AB - Human cytomegalovirus (HCMV), a betaherpesvirus, has developed several ways to evade the immune system, notably downregulation of cell surface expression of major histocompatibility complex class I heavy chains. Here we report that HCMV has devised another means to compromise immune surveillance mechanisms. Extracellular accumulation of both constitutively produced monocyte chemoattractant protein (MCP)-1 and tumor necrosis factor-superinduced RANTES (regulated on activation, normal T cell expressed and secreted) was downregulated in HCMV-infected fibroblasts in the absence of transcriptional repression or the expression of polyadenylated RNA for the cellular chemokine receptors CCR-1, CCR 3, and CCR-5. Competitive binding experiments demonstrated that HCMV-infected cells bind RANTES, MCP-1, macrophage inflammatory protein (MIP)-1beta, and MCP-3, but not MCP-2, to the same receptor as does MIP-1alpha, which is not expressed in uninfected cells. HCMV encodes three proteins with homology to CC chemokine receptors: US27, US28, and UL33. Cells infected with HCMV mutants deleted of US28, or both US27 and US28 genes, failed to downregulate extracellular accumulation of either RANTES or MCP-1. In contrast, cells infected with a mutant deleted of US27 continues to bind and downregulate those chemokines. Depletion of chemokines from the culture medium was at least partially due to continuous internalization of extracellular chemokine, since exogenously added, biotinylated RANTES accumulated in HCMV-infected cells. Thus, HCMV can modify the chemokine environment of infected cells through intense sequestering of CC chemokines, mediated principally by expression of the US28-encoded chemokine receptor. PMID- 9730888 TI - CD161 (NKR-P1A) costimulation of CD1d-dependent activation of human T cells expressing invariant V alpha 24 J alpha Q T cell receptor alpha chains. AB - A population of human T cells expressing an invariant V alpha 24 J alpha Q T cell antigen receptor (TCR) alpha chain and high levels of CD161 (NKR-P1A) appears to play an immunoregulatory role through production of both T helper (Th) type 1 and Th2 cytokines. Unlike other CD161(+) T cells, the major histocompatibility complex-like nonpolymorphic CD1d molecule is the target for the TCR expressed by these T cells (V alpha 24(invt) T cells) and by the homologous murine NK1 (NKR P1C)+ T cell population. In this report, CD161 was shown to act as a specific costimulatory molecule for TCR-mediated proliferation and cytokine secretion by V alpha 24(invt) T cells. However, in contrast to results in the mouse, ligation of CD161 in the absence of TCR stimulation did not result in V alpha 24(invt) T cell activation, and costimulation through CD161 did not cause polarization of the cytokine secretion pattern. CD161 monoclonal antibodies specifically inhibited V alpha 24(invt) T cell proliferation and cytokine secretion in response to CD1d+ target cells, demonstrating a physiological accessory molecule function for CD161. However, CD1d-restricted target cell lysis by activated V alpha 24(invt) T cells, which involved a granule-mediated exocytotic mechanism, was CD161 independent. In further contrast to the mouse, the signaling pathway involved in V alpha 24(invt) T cell costimulation through CD161 did not appear to involve stable association with tyrosine kinase p56(Lck). These results demonstrate a role for CD161 as a novel costimulatory molecule for TCR-mediated recognition of CD1d by human V alpha 24(invt) T cells. PMID- 9730890 TI - Antiinflammatory effects of CD95 ligand (FasL)-induced apoptosis. AB - Apoptosis is critical to homeostasis of multicellular organisms. In immune privileged sites such as the eye, CD95 ligand (FasL)-induced apoptosis controls dangerous inflammatory reactions that can cause blindness. Recently, we demonstrated that apoptotic cell death of inflammatory cells was a prerequisite for the induction of immune deviation after antigen presentation in the eye. In this report, we examine the mechanism by which this takes place. Our results show that Fas- mediated apoptosis of lymphoid cells leads to rapid production of interleukin (IL)-10 in these cells. The apoptotic cells containing IL-10 are responsible for the activation of immune deviation through interaction with antigen-presenting cells (APC). In support of this, we found that apoptotic cells from IL-10(+/+) animals fed to APC in vitro promote Th2 cell differentiation, whereas apoptotic IL-10(-/-) cells, as well as nonapoptotic cells, favor Th1 induction. Thus, apoptotic cell death and tolerance are linked through the production of an antiinflammatory cytokine to prevent dangerous and unwanted immune responses that might compromise organ integrity. PMID- 9730889 TI - The specificity of peptides bound to human histocompatibility leukocyte antigen (HLA)-B27 influences the prevalence of arthritis in HLA-B27 transgenic rats. AB - Human histocompatibility leukocyte antigen B27 is highly associated with the rheumatic diseases termed spondyloarthropathies, but the mechanism is not known. B27 transgenic rats develop a spontaneous disease resembling the human spondyloarthropathies that includes arthritis and colitis. To investigate whether this disease requires the binding of specific peptides to B27, we made a minigene construct in which a peptide from influenza nucleoprotein, NP383-391 (SRYWAIRTR), which binds B27 with high affinity, is targeted directly to the ER by the signal peptide of the adenovirus E3/gp19 protein. Rats transgenic for this minigene, NP1, were made and bred with B27 rats. The production of the NP383-391 peptide in B27(+)NP1(+) rats was confirmed immunologically and by mass spectrometry. The NP1 product displaced approximately 90% of the 3H-Arg-labeled endogenous peptide fraction in B27(+)NP1(+) spleen cells. Male B27(+)NP1(+) rats had a significantly reduced prevalence of arthritis, compared with B27(+)NP- males or B27(+) males with a control construct, NP2, whereas colitis was not significantly affected by the NP1 transgene. These findings support the hypothesis that B27-related arthritis requires binding of a specific peptide or set of peptides to B27, and they demonstrate a method for efficient transgenic targeting of peptides to the ER. PMID- 9730891 TI - Highly restricted T cell repertoire shaped by a single major histocompatibility complex-peptide ligand in the presence of a single rearranged T cell receptor beta chain. AB - The T cell repertoire is shaped by positive and negative selection of thymocytes through the interaction of alpha/beta-T cell receptors (TCR) with self-peptides bound to self-major histocompatibility complex (MHC) molecules. However, the involvement of specific TCR-peptide contacts in positive selection remains unclear. By fixing TCR-beta chains with a single rearranged TCR-beta irrelevant to the selecting ligand, we show here that T cells selected to mature on a single MHC-peptide complex express highly restricted TCR-alpha chains in terms of Valpha usage and amino acid residue of their CDR3 loops, whereas such restriction was not observed with those selected by the same MHC with diverse sets of self peptides including this peptide. Thus, we visualized the TCR structure required to survive positive selection directed by this single ligand. Our findings provide definitive evidence that specific recognition of self-peptides by TCR could be involved in positive selection of thymocytes. PMID- 9730892 TI - Efficient peripheral clonal elimination of B lymphocytes in MRL/lpr mice bearing autoantibody transgenes. AB - Peripheral B cell tolerance was studied in mice of the autoimmune-prone, Fas deficient MRL/ lpr.H-2(d) genetic background by introducing a transgene that directs expression of membrane-bound H-2Kb antigen to liver and kidney (MT-Kb) and a second transgene encoding antibody reactive with this antigen (3-83mu delta, anti-Kk,b). Control immunoglobulin transgenic (Ig-Tg) MRL/lpr.H-2(d) mice lacking the Kb antigen had large numbers of splenic and lymph node B cells bearing the transgene-encoded specificity, whereas B cells of the double transgenic (Dbl-Tg) MRL/lpr.H-2(d) mice were deleted as efficiently as in Dbl-Tg mice of a nonautoimmune B10.D2 genetic background. In spite of the severely restricted peripheral B cell repertoire of the Ig-Tg MRL/lpr.H-2(d) mice, and notwithstanding deletion of the autospecific B cell population in the Dbl-Tg MRL/lpr.H-2(d) mice, both types of mice developed lymphoproliferation and exhibited elevated levels of IgG anti-chromatin autoantibodies. Interestingly, Dbl-Tg MRL/lpr.H-2(d) mice had a shorter lifespan than Ig-Tg MRL/lpr.H-2(d) mice, apparently as an indirect result of their relative B cell lymphopenia. These data suggest that in MRL/lpr mice peripheral B cell tolerance is not globally defective, but that certain B cells with receptors specific for nuclear antigens are regulated differently than are cells reactive to membrane autoantigens. PMID- 9730894 TI - p56lck signals for regulating thymocyte development can be distinguished by their dependency on Rho function. AB - The tyrosine kinase p56lck regulates the differentiation and proliferative expansion of pre-T cells. However, nothing is known about other signaling molecules that operate with p56lck to mediate the pleiotropic changes that occur at this stage of thymocyte development. We used a genetic strategy to examine the requirement for the GTPase Rho in p56lck-mediated signals in the thymus. By generating mice double transgenic for a constitutively activated form of p56lck (p56lckF505) and the Rho inhibitor C3 transferase we were able to compare thymocyte development in mice expressing active p56lck on a wild-type or Rho- background. Thymocytes expressing active p56lck show enhanced proliferation of pre-T cells resulting in increased numbers of late pre-T cells, however, this dramatic effect on pre-T cell proliferation is lost when the p56lck transgene is expressed in thymocytes lacking endogenous Rho GTPase function. Expression of active p56lck also generates double positive (DP) thymocytes with low levels of CD2 antigen expression. Again, p56lck cannot prevent expression of CD2 when expressed on a Rho- background. CD4(+)CD8(+) DP cells expressing active p56lck have been shown to lack functional alpha/beta-T cell receptor (TCR) complexes due to p56lck-mediated inhibition of TCR gene Vbeta-Dbeta rearrangement. This inhibition of TCR expression by active p56lck is unimpaired in the absence of Rho function. The signaling pathways that are mediated by p56lck and control thymocyte proliferation, alpha/beta-TCR and CD2 antigen expression can thus be distinguished by their dependency on Rho function. PMID- 9730893 TI - Dual signaling of the Fas receptor: initiation of both apoptotic and necrotic cell death pathways. AB - Murine L929 fibrosarcoma cells were transfected with the human Fas (APO-1/CD95) receptor, and the role of various caspases in Fas-mediated cell death was assessed. Proteolytic activation of procaspase-3 and -7 was shown by Western analysis. Acetyl-Tyr-Val-Ala-Asp-chloromethylketone and benzyloxycarbonyl Asp(OMe)-Glu(OMe)-Val-Asp(OMe)-fluoromethylketone++ +, tetrapeptide inhibitors of caspase-1- and caspase-3-like proteases, respectively, failed to block Fas induced apoptosis. Unexpectedly, the broad-spectrum caspase inhibitors benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethylketone and benzyloxycarbonyl Asp(OMe)-fluoromethylketone rendered the cells even more sensitive to Fas mediated cell death, as measured after 18 h incubation. However, when the process was followed microscopically, it became clear that anti-Fas-induced apoptosis of Fas-transfected L929 cells was blocked during the first 3 h, and subsequently the cells died by necrosis. As in tumor necrosis factor (TNF)-induced necrosis, Fas treatment led to accumulation of reactive oxygen radicals, and Fas-mediated necrosis was inhibited by the oxygen radical scavenger butylated hydroxyanisole. However, in contrast to TNF, anti-Fas did not activate the nuclear factor kappaB under these necrotic conditions. These results demonstrate the existence of two different pathways originating from the Fas receptor, one rapidly leading to apoptosis, and, if this apoptotic pathway is blocked by caspase inhibitors, a second directing the cells to necrosis and involving oxygen radical production. PMID- 9730896 TI - Molecular cloning of NKp46: a novel member of the immunoglobulin superfamily involved in triggering of natural cytotoxicity. AB - NKp46 has been shown to represent a novel, natural killer (NK) cell-specific surface molecule, involved in human NK cell activation. In this study, we further analyzed the role of NKp46 in natural cytotoxicity against different tumor target cells. We provide direct evidence that NKp46 represents a major activating receptor involved in the recognition and lysis of both human and murine tumor cells. Although NKp46 may cooperate with other activating receptors (including the recently identified NKp44 molecule) in the induction of NK-mediated lysis of human tumor cells, it may represent the only human NK receptor involved in recognition of murine target cells. Molecular cloning of the cDNA encoding the NKp46 molecule revealed a novel member of the immunoglobulin (Ig) superfamily, characterized by two C2-type Ig-like domains in the extracellular portion. The transmembrane region contains the positively charged amino acid Arg, which is possibly involved in stabilizing the association with CD3zeta chain. The cytoplasmic portion, spanning 30 amino acids, does not contain immunoreceptor tyrosine-based activating motifs. Analysis of a panel of human/hamster somatic cell hybrids revealed segregation of the NKp46 gene on human chromosome 19. Assessment of the NKp46 mRNA expression in different tissues and cell types unambiguously confirmed the strict NK cell specificity of the NKp46 molecule. Remarkably, in line with the ability of NKp46 to recognize ligand(s) on murine target cells, the cDNA encoding NKp46 was found to be homologous to a cDNA expressed in murine spleen. In conclusion, this study reports the first characterization of the molecular structure of a NK-specific receptor involved in the mechanism of NK cell activation during natural cytotoxicity. PMID- 9730898 TI - Qa-1b binds conserved class I leader peptides derived from several mammalian species. AB - Qa-1b binds a peptide (AMAPRTLLL), referred to as Qdm (for Qa-1 determinant modifier), derived from the signal sequence of murine class Ia molecules. This peptide binds with high affinity and accounts for almost all of the peptides associated with this molecule. Human histocompatibility leukocyte antigen (HLA) E, a homologue of Qa-1b, binds similar peptides derived from human class Ia molecules and interacts with CD94/NKG2 receptors on natural killer cells. We used surface plasmon resonance to determine the ability of Qa-1b to bind related ligands representing peptides derived from the leaders of class I molecules from several mammalian species. All of the peptides reported to bind HLA-E bound readily to Qa-1b. In addition, peptides derived from leader segments of different mammals also bound to Qa-1b, indicating a conservation of this "Qdm-like" epitope throughout mammalian evolution. We have attempted to define a minimal peptide on a polyglycine backbone that binds Qa-1b. Our previous findings showed that P2 and P9 are important but not sufficient for binding to Qa-1b. Although a minimum peptide (GMGGGGLLL) bound Qa-1(b), its interaction was relatively weak, as were peptides sharing five or six residues with Qdm, indicating that multiple native residues are required for a strong interaction. This finding is consistent with the observation that this molecule preferentially binds this single ligand. PMID- 9730895 TI - Gonococcal invasion of epithelial cells driven by P.IA, a bacterial ion channel with GTP binding properties. AB - The neisserial porin P.I is a GTP binding protein that forms a voltage-gated channel that translocates into mammalian cell membranes and modulates host cell signaling events. Here, we report that P.I confers invasion of the bacterial pathogen Neisseria gonorrhoeae into Chang epithelial cells and that this event is controlled by GTP, as well as other phosphorus-containing compounds. Bacterial invasion was observed only for strains carrying the P.IA subtype of porin, which is typically associated with the development of disseminated neisserial disease, and did not require opacity outer membrane proteins, previously recognized as gonococcal invasins. Allelic replacement studies showed that bacterial invasiveness cotransferred with the P.IA (por1A) gene. Mutation of the P.I associated protein Rmp did not alter the invasive properties. Cross-linking of labeled GTP to the porin revealed more efficient GTP binding to the P.IA than P.IB porin subtype. GTP binding was inhibited by an excess of unlabeled GTP, ATP, and GDP, as well as inorganic phosphate, but not by UTP or beta-glycerophosphate, fully in line with the respective invasion-inhibitory activities observed for these compounds. The P.IA-mediated cellular invasion may explain the more invasive behavior of P.IA strains in the natural infection and may broaden the basis for the development of a P.I-based gonococcal vaccine. PMID- 9730897 TI - Transporters associated with antigen processing (TAP)-independent presentation of soluble insulin to alpha/beta T cells by the class Ib gene product, Qa-1(b). AB - T cell hybridomas isolated from nonresponder H-2(b) mice immunized with pork insulin were stimulated by insulin in the presence of major histocompatibility complex (MHC)-unmatched antigen presenting cells. The restriction element used by these CD4(-) T cells was mapped to an oligomorphic MHC class Ib protein encoded in the T region and identified as Qa-1(b) using transfectants. The antigenic determinant was localized to the insulin B chain, and experiments with truncated peptides suggested that it is unexpectedly long, comprising most or all of the 30 amino acid B chain. The antigen processing pathway used to present insulin to the Qa-1(b)- restricted T cells does not require transporters associated with antigen processing (TAP), and it is inhibited by chloroquine. A wide variety of cell lines from different tissues efficiently present soluble insulin to Qa-1(b) restricted T cells, and insulin presentation is not enhanced by phagocytic stimuli. Our results demonstrate that Qa-1(b) can function to present exogenous protein to T cells in a manner similar to MHC class II molecules. Therefore, this class Ib protein may have access to a novel antigen processing pathway that is not available to class Ia molecules. PMID- 9730899 TI - Differential regulation and ATP requirement for caspase-8 and caspase-3 activation during CD95- and anticancer drug-induced apoptosis. AB - Apoptosis is induced by different stimuli, among them triggering of the death receptor CD95, staurosporine, and chemotherapeutic drugs. In all cases, apoptosis is mediated by caspases, although it is unclear how these diverse apoptotic stimuli cause protease activation. Two regulatory pathways have been recently identified, but it remains unknown whether they are functionally independent or linked to each other. One is mediated by recruitment of the proximal regulator caspase-8 to the death receptor complex. The other pathway is controlled by the release of cytochrome c from mitochondria and the subsequent ATP-dependent activation of the death regulator apoptotic protease-activating factor 1 (Apaf 1). Here, we report that both pathways can be dissected by depletion of intracellular ATP. Prevention of ATP production completely inhibited caspase activation and apoptosis in response to chemotherapeutic drugs and staurosporine. Interestingly, caspase-8, whose function appeared to be restricted to death receptors, was also activated by these drugs under normal conditions, but not after ATP depletion. In contrast, inhibition of ATP production did not affect caspase activation after triggering of CD95. These results suggest that chemotherapeutic drug-induced caspase activation is entirely controlled by a receptor-independent mitochondrial pathway, whereas CD95-induced apoptosis can be regulated by a separate pathway not requiring Apaf-1 function. PMID- 9730900 TI - Interleukin 6 dependence of anti-DNA antibody production: evidence for two pathways of autoantibody formation in pristane-induced lupus. AB - Pristane induces a lupus-like syndrome in nonautoimmune mice characterized by the development of glomerulonephritis and lupus-associated autoantibodies. This is accompanied by overproduction of interleukin (IL)-6, a cytokine linked with autoimmune phenomena. The goal of this study was to evaluate the role of IL-6 in autoantibody production in pristane-induced lupus. BALB/cAn IL-6-deficient (-/-) and -intact (+/+) mice were treated with pristane or phosphate-buffered saline, and autoantibody production was evaluated. Pristane induced high levels of immunoglobulin (Ig)G anti-single-stranded DNA, -double-stranded (ds)DNA, and chromatin antibodies in IL-6(+/+), but not IL-6(-/-) mice by enzyme-linked immunosorbent assay. High titer IgG anti-dsDNA antibodies also were detected in sera from +/+, but not -/-, mice by Crithidia luciliae kinetoplast staining. The onset of IgG anti-dsDNA antibody production in +/+ mice occurred >5 mo after pristane treatment, well after the onset of nephritis, suggesting that these antibodies are not directly responsible for inducing renal disease. In contrast to anti-DNA, the frequencies of anti-nRNP/Sm and anti-Su antibodies were similar in pristane-treated IL-6(-/-) and IL-6(+/+) mice. However, levels were higher in the +/+ group. These results suggest that IgG anti-DNA and chromatin antibodies in pristane-treated mice are strictly IL-6 dependent, whereas induction of anti nRNP/Sm and Su autoantibodies is IL-6 independent. The IL-6 dependence of anti DNA, but not anti-nRNP/Sm, may have implications for understanding the patterns of autoantibody production in lupus. Anti-DNA antibodies are produced transiently, mainly during periods of disease activity, whereas anti-nRNP/Sm antibody levels are relatively insensitive to disease activity. This may reflect the differential IL-6 dependence of the two responses. PMID- 9730901 TI - Paired immunoglobulin-like receptor (PIR)-A is involved in activating mast cells through its association with Fc receptor gamma chain. AB - Paired immunoglobulin-like receptor (PIR)-A and PIR-B possess similar ectodomains with six immunoglobulin-like loops, but have distinct transmembrane and cytoplasmic domains. PIR-B bears immunoreceptor tyrosine-based inhibitory motif (ITIM) sequences in its cytoplasmic domain that recruit Src homology (SH)2 domain containing tyrosine phosphatases SHP-1 and SHP-2, leading to inhibition of B and mast cell activation. In contrast, the PIR-A protein has a charged Arg residue in its transmembrane region and a short cytoplasmic domain that lacks ITIM sequences. Here we show that Fc receptor gamma chain, containing an immunoreceptor tyrosine-based activation motif (ITAM), associates with PIR-A. Cross-linking of this PIR-A complex results in mast cell activation such as calcium mobilization in an ITAM-dependent manner. Thus, our data provide evidence for the existence of two opposite signaling pathways upon PIR aggregation. PIR-A induces the stimulatory signal by using ITAM in the associated gamma chain, whereas PIR-B mediates the inhibitory signal through its ITIMs. PMID- 9730902 TI - TRANCE is necessary and sufficient for osteoblast-mediated activation of bone resorption in osteoclasts. AB - TRANCE (tumor necrosis factor-related activation-induced cytokine) is a recently described member of the tumor necrosis factor superfamily that stimulates dendritic cell survival and has also been found to induce osteoclastic differentiation from hemopoietic precursors. However, its effects on mature osteoclasts have not been defined. It has long been recognized that stimulation of osteoclasts by agents such as parathyroid hormone (PTH) occurs through a hormonal interaction with osteoblastic cells, which are thereby induced to activate osteoclasts. To determine whether TRANCE accounts for this activity, we tested its effects on mature osteoclasts. TRANCE rapidly induced a dramatic change in osteoclast motility and spreading and inhibited apoptosis. In populations of osteoclasts that were unresponsive to PTH, TRANCE caused activation of bone resorption equivalent to that induced by PTH in the presence of osteoblastic cells. Moreover, osteoblast-mediated stimulation of bone resorption was abrogated by soluble TRANCE receptor and by the soluble decoy receptor osteoprotegerin (OPG), and stimulation of isolated osteoclasts by TRANCE was neutralized by OPG. Thus, TRANCE expression by osteoblasts appears to be both necessary and sufficient for hormone-mediated activation of mature osteoclasts, and TRANCE-R is likely to be a receptor for signal transduction for activation of the osteoclast and its survival. PMID- 9730903 TI - D1 dopamine receptor agonists mediate activation of p38 mitogen-activated protein kinase and c-Jun amino-terminal kinase by a protein kinase A-dependent mechanism in SK-N-MC human neuroblastoma cells. AB - We investigated the effects of D1 dopamine receptor stimulation on the activation of mitogen-activated protein kinases (MAPKs) in SK-N-MC human neuroblastoma cells. We found that the D1 dopamine receptor agonist SKF38393 induced similar time- and dose-related activation of p38 MAPK and c-Jun amino-terminal kinase (JNK), whereas extracellular signal-regulated kinase activity was not affected by D1 dopamine receptor stimulation. Maximal stimulation of p38 MAPK and JNK was observed after a 15-min incubation with 100 microM SKF38393. In contrast, 10 microM quinpirole, a D2 dopamine receptor agonist, did not activate p38 MAPK or JNK. Treatment of cells with 10 muM SCH23390, a D1 dopamine receptor antagonist, significantly inhibited the activation of both kinases by SKF38393. These results indicate that activation of the p38 MAPK and JNK signaling pathways is mediated by dopamine D1 receptors in SK-N-MC neuroblastoma cells. Furthermore, dibutyryl cAMP mimicked SKF38393-mediated stimulation of p38 MAPK and JNK. Inhibition of protein kinase A by 1 microM H-89 or 10 microM adenosine 3', 5'-cyclic monophosphothioate (Rp-isomer, triethylammonium salt) markedly attenuated the activation of p38 MAPK and JNK. Conversely, the selective protein kinase C inhibitor calphostin C did not block D1 dopamine receptor-stimulated activation of p38 MAPK and JNK. These results demonstrate, for the first time, that the Gs coupled D1 dopamine receptor activates the p38 MAPK and JNK signaling pathways by a protein kinase A-dependent mechanism. PMID- 9730904 TI - Cannabinoid receptor agonists protect cultured rat hippocampal neurons from excitotoxicity. AB - Cannabinoid receptor agonists act presynaptically to inhibit the release of glutamate. Because other drugs with this action are known to reduce excitotoxicity, we tested several cannabimimetics in a model of synaptically mediated neuronal death. Reduction of the extracellular Mg2+ concentration to 0.1 mM evoked a repetitive pattern of intracellular Ca2+ concentration ([Ca2+]i) spiking that, when maintained for 24 hr, resulted in significant neuronal death. The [Ca2+]i spiking and cell death in this model result from excessive activation of N-methyl-D-aspartate receptors, as indicated by the inhibition of both [Ca2+]i spiking and neuronal death by the N-methyl-D-aspartate receptor antagonist CGS19755 (10 microM). The cannabimimetic drug Win55212-2 (100 nM) completely blocked [Ca2+]i spiking and prevented neuronal death induced by low extracellular Mg2+ concentrations. These effects on [Ca2+]i spiking and viability were stereoselective and were prevented by the CB1 receptor antagonist SR141716 (100 nM). The partial agonist CP55940 (100 nM) also afforded significant protection from excitotoxicity. Cannabimimetic drugs did not protect cells from the direct application of glutamate (30 microM). These data suggest that cannabimimetic drugs may slow the progression of neurodegenerative diseases. PMID- 9730905 TI - Down-regulation of cytochrome P450 2C family members and positive acute-phase response gene expression by peroxisome proliferator chemicals. AB - In this study, we show that peroxisome proliferator chemical (PPC) exposure leads to alterations in the expression of genes in rat liver regulated by the sex specific growth hormone secretory pattern and induced during inflammation. Expression of the male-specific cytochrome P450 (P450) 2C11 and alpha2 urinary globulin (alpha2u) genes and the female-specific P450 2C12 gene was down regulated by some PPC. Expression of P450 2C13, also under control by the sex specific growth hormone secretory pattern, was not altered by PPC treatment, indicating that regulation of CYP2C family members does not involve perturbation of the growth hormone secretory pattern. In contrast to the increases in expression observed when inflammation was induced in male rats, two positive acute-phase response genes, alpha1-acid glycoprotein and beta-fibrinogen, were decreased by PPC exposure. The down-regulation of the P450 2C11 by WY-14,643 could be reproduced in cultured rat hepatocytes, indicating the down-regulation is a direct effect. Experiments in wild-type mice and mice that lacked a functional peroxisome proliferator-activated receptor-alpha gene showed that down regulation by WY of alpha1-acid glycoprotein, beta-fibrinogen, and a mouse homologue of alpha2u was dependent on peroxisome proliferator-activated receptor alpha expression. Our results demonstrate that PPC exposure leads to down regulation of diverse liver-specific genes, including CYP2C family members important in hormonal homeostasis and acute-phase response genes important in inflammatory responses. PMID- 9730906 TI - Regulation of rat hepatic cytochrome P450 expression by sterol biosynthesis inhibition: inhibitors of squalene synthase are potent inducers of CYP2B expression in primary cultured rat hepatocytes and rat liver. AB - The effects of treatment with squalestatin 1, a potent inhibitor of squalene synthase, the first committed enzyme of sterol biosynthesis, were examined on cytochrome P450 expression in primary cultured rat hepatocytes and rat liver. Incubation of cultured hepatocytes with squalestatin 1 caused marked accumulations (maximal elevations that were approximately 25-100% of phenobarbital-elicited increases) of CYP2B mRNA and immunoreactive protein but not of CYP1A, CYP3A, or CYP4A. Squalestatin 1 treatment increased CYP2B and 3 hydroxy-3-methylglutaryl coenzyme A reductase mRNA content in hepatocyte cultures with comparable potencies (ED50 = 5.0 and 18 nM, respectively), and significantly induced CYP2B (mRNA, immunoreactive protein, and pentoxyresorufin O-dealkylase activity) in the livers of treated rats, producing maximal increases at a dose of 25 mg/kg/day that were approximately 32-87% of phenobarbital-induced increases. Squalestatin 1 treatment induced both CYP2B1 and CYP2B2 and activated reporter gene expression in cultured hepatocytes transiently transfected with a plasmid containing approximately 2.4 kb of CYP2B1 gene 5'-flanking region or containing a previously described phenobarbital-responsive region. Coincubation of cultured hepatocytes with 25-hydroxycholesterol suppressed squalestatin 1-mediated CYP2B and 3-hydroxy-3-methylglutaryl coenzyme A mRNA induction with approximately the same potency. Treatment of cultures with SQ-34919, a structurally distinct squalene synthase inhibitor, produced the same selective CYP2B mRNA induction as did squalestatin 1. These results suggest that inhibition of hepatic sterol synthesis activates processes that culminate in increased CYP2B gene transcription. PMID- 9730907 TI - Agonist-induced internalization of the P2Y2 receptor. AB - The Gq/phospholipase C-linked human P2Y2 receptor was tagged at its amino terminus with the hemagglutinin A (HA) epitope sequence (P2Y2-HA) and stably expressed in 1321N1 human astrocytoma cells. Neither the pharmacological selectivity nor the signaling properties of the receptor were altered by the presence of the epitope. An enzyme-linked immunosorbent assay was developed to quantify cell surface levels of P2Y2-HA receptors using an anti-HA antibody. Incubation of cells with P2Y2 receptor agonists resulted in a concentration of agonist- and time-dependent decrease in cell surface immunoreactivity. Methodology for indirect immunofluorescence confocal microscopy was developed and applied to demonstrate that the agonist-promoted decreases in cell surface immunoreactivity paralleled increases in intracellular immunoreactivity. Agonist induced internalization of P2Y2 receptors was demonstrated directly by prelabeling P2Y2-HA receptors with antibody before agonist challenge and then quantifying the movement of receptors from a cell surface to intracellular localization in the presence of agonist. Removal of agonist from the medium resulted in recovery of cell surface immunoreactivity to control levels within approximately 1 hr. Incubation of P2Y2-HA receptor-expressing cells with P2Y2 receptor agonists also resulted in receptor-specific desensitization of nucleotide-promoted inositol phosphate accumulation. This loss of responsiveness occurred more rapidly and to a greater extent than did the agonist-promoted loss of surface receptors. Inhibition of receptor internalization by reduction of temperature to 16 degrees had no effect on the capacity of nucleotides to induce P2Y2 receptor-specific desensitization. These results illustrate that the P2Y2 receptor undergoes agonist-promoted movement to an intracellular compartment. This receptor internalization is not required for agonist-induced desensitization. PMID- 9730908 TI - Transgenic animals with inducible, targeted gene expression in brain. AB - Several inducible gene expression systems have been developed in vitro in recent years to overcome limitations with traditional transgenic mice. One of these, the tetracycline-regulated system, has been used successfully in vivo. Nevertheless, concerns remain about the ability of this system to direct high levels of transgene expression in vivo and to enable such expression to be turned on and off effectively. We report here the generation, using a modified tetracycline regulated system under the control of the neuron-specific enolase promoter, of several lines of mice that direct transgene expression to specific brain regions, including the striatum, cerebellum, CA1 region of the hippocampus, or deep layers of cerebral neocortex. Transgene expression in these mice can be turned off completely with low doses of doxycycline (a tetracycline derivative) and driven to very high levels in the absence of doxycycline. We demonstrate this tissue specific, inducible expression for three transgenes: those that encode luciferase (a reporter protein) or DeltaFosB or the cAMP-response element binding protein (CREB) (two transcription factors). The various lines of transgenic mice demonstrate an inducible system that generates high levels of transgene expression in specific brain regions and represent novel and powerful tools with which to study the functioning of these (or potentially any other) genes in the brain. PMID- 9730909 TI - Nutritional status modulates rat liver cytochrome P450 arachidonic acid metabolism. AB - Alterations in nutritional status affect hepatic cytochrome P450 levels. Since cytochromes P450 participate in the metabolism of arachidonic acid, we hypothesized that changes in liver P450 arachidonic acid metabolism occur during fasting and refeeding. Male Fisher 344 rats were either fed, fasted 48 hr (F48), fasted 48 hr and then refed 6 hr (F48/R6), or fasted 48 hr and then refed 24 hr (F48/R24). F48 rats had reduced body weight, increased plasma beta hydroxybutyrate, and reduced plasma insulin compared with the other groups. Although there was no significant change in total liver P450 content, there was a significant 20%, 48%, and 24% reduction in total hepatic microsomal arachidonic acid metabolism in F48, F48/R6, and F48/R24 rats, respectively, compared with fed rats. Epoxygenase activity decreased by 28%, 51%, and 26% in F48, F48/R6, and F48/R24 rats, respectively. In contrast, omega-1 hydroxylase activity increased by 126% in F48 rats compared with fed rats. Immunoblotting revealed that levels of CYP2C11 protein were markedly reduced, whereas levels of CYP2E1 protein were markedly increased in the F48 and F48/R6 groups. In contrast, levels of CYP1A1, CYP1A2, CYP2B1, CYP2J3, CYP4A1, and CYP4A3 were unchanged with fasting/refeeding. Northern blots revealed that levels of CYP2C11 mRNAs were decreased, whereas CYP2E1 mRNAs were increased in F48 and F48/R6 rats. Recombinant CYP2C11 metabolized arachidonic acid primarily to epoxides with preference for the 14(S),15(R)-, 11(R), 12(S)-, and 8(S),9(R)- epoxyeicosatrienoic acid enantiomers. We conclude that (1) nutritional status affects hepatic microsomal arachidonic acid metabolism, (2) reduced epoxygenase activity in F48 and F48/R6 rats is accompanied by decreased levels of CYP2C11, (3) increased omega-1 hydroxylase activity is accompanied by augmented levels of CYP2E1, and (4) the effects of fasting on CYP2C11 and CYP2E1 expression occur at the pretranslational level. PMID- 9730910 TI - Inhibition of cyclic AMP-dependent kinase by expression of a protein kinase inhibitor/enhanced green fluorescent fusion protein attenuates angiotensin II induced type 1 AT1 receptor mRNA down-regulation in vascular smooth muscle cells. AB - Expression of the angiotensin II type 1 receptor (AT1-R) mRNA in vascular smooth muscle cells (VSMC) is down-regulated by a variety of agonists, including growth factors, agonists of Galphaq protein-coupled receptors, and activators of adenylyl cyclase. To determine whether cAMP-dependent protein kinases (PKA) participates in AT1-R mRNA down-regulation controlled by multiple classes of receptors, a PKA inhibitor peptide (PKIalpha) was developed and expressed in rat VSMC as a fusion with the enhanced green fluorescent protein (eGFP). PKA activity elicited both by forskolin and angiotensin II is suppressed in cells expressing this fusion protein (PKIalpha-eGFP), but platelet-derived growth factor-BB does not stimulate PKA activity in this preparation. PKIalpha-eGFP expression fully inhibits the forskolin-stimulated down-regulation of AT1-R mRNA levels and blocks 50% of the effect elicited by angiotensin II. This indicates that PKA plays a substantial role in angiotensin II-stimulated AT1-R mRNA down-regulation. However, inhibition of PKA has no effect on AT1-R mRNA down-regulation caused by platelet-derived growth factor-BB. These findings show how agonists such as angiotensin II that are not normally considered as activators of PKA can use PKA dependent processes to modulate gene expression. These findings also provide definitive evidence that PKA-dependent pathways are involved in modulation of AT1 R mRNA levels in VSMC. PMID- 9730911 TI - A dileucine sequence and an upstream glutamate residue in the intracellular carboxyl terminus of the vasopressin V2 receptor are essential for cell surface transport in COS.M6 cells. AB - Little is known concerning the intracellular transport of the G protein-coupled receptors (GPCRs). Previous studies suggested a functional role for those residues immediately preceding the conserved palmitoylated cysteine residues in the intracellular carboxyl termini of some GPCRs in cell surface transport. For the human vasopressin V2 receptor, we assessed the significance of a dileucine sequence with an upstream glutamate residue (ELRSLLCC) in mediating cell surface delivery. A series of deletion and point mutants in this region were constructed, and the mutant receptors were expressed in transiently transfected COS.M6 cells. By using [3H]arginine vasopressin binding assays to intact cells and immunofluorescence studies with intact and permeabilized cells, we show that residues E335 (mutant E335Q) and L339 (mutant L339T) are obligatory for receptor transport to the plasma membrane. Residue L340 has a minor but significant influence. [3H]Arginine vasopressin binding experiments on membranes of lysed cells failed to detect any intracellular binding sites for the transport deficient mutant receptors, suggesting that residues E335 and L339 participate in receptor folding. Studies with green fluorescent protein-tagged receptors demonstrate that the bulk of the mutant receptors E335Q and L339T are trapped in the endoplasmic reticulum. Complex glycosylation was absent in these mutant receptors, supporting this conclusion. These data demonstrate that the glutamate/dileucine motif of the vasopressin V2 receptor is critical for the escape of the receptor from the endoplasmic reticulum, most presumably by establishing a functional and transport-competent folding state. A databank analysis revealed that these residues are part of a conserved region in the GPCR family. PMID- 9730912 TI - Constitutive cyclooxygenase-2 expression in healthy human and rabbit gastric mucosa. AB - Selective cyclooxygenase (COX)-2 inhibitors are expected to cause fewer gastric side effects because of sparing of COX-1-dependent prostaglandin (PG) synthesis in the gastric mucosa. However, the possible contribution of COX-2 to overall gastric PG biosynthesis is not known. This study demonstrates constitutive expression of COX-2 mRNA and protein in apparently healthy human and rabbit gastric mucosa. This basal expression of COX-2 protein in human gastric mucosa was increased by lipopolysaccharide and phorbol ester, indicating its up regulation in response to appropriate stimuli. The functional significance of COX 2-dependent PG formation was studied in terms of PGE2 generation in the rabbit mucosa and its inhibition by the COX-2-selective inhibitor flosulide. There was concentration-dependent (IC50 = 107 +/- 55 nM) and ultimately complete inhibition of PGE2 generation by flosulide. In addition, gastric mucosa generated 15 hydroxyeicosatetraenoic acid upon treatment with acetylsalicylic acid. The data suggest an important role for COX-2-dependent PG production in apparently healthy gastric mucosa and raise the issue of whether selective COX-2 inhibitors might also interfere with physiological PG formation and actions in the stomach. PMID- 9730913 TI - Evidence that the p2y3 receptor is the avian homologue of the mammalian P2Y6 receptor. AB - A P2Y receptor with 65% identity to mammalian P2Y6 receptors, termed the p2y3 receptor, was recently cloned from a chick brain cDNA library and was proposed to represent a novel P2Y receptor subtype [Mol Pharmacol 50:258-265 (1996)]. We cloned the turkey homologue of the chick p2y3 receptor, which shares high sequence identity (97.6%) with the chick receptor, and we stably expressed this receptor and the rat P2Y6 receptor in 1321N1 human astrocytoma cells. The capacities of uridine and adenine nucleotides to promote inositol phosphate accumulation and intracellular Ca2+ mobilization were determined for both receptors. UDP and 5-bromo-UDP were the most potent agonists and UTP was a less potent full agonist at both receptors. In contrast, adenine nucleotides and nucleotide derivatives were relatively more potent at the turkey p2y3 receptor than at the rat P2Y6 receptor. To determine whether the avian p2y3 receptor defined a new subtype of mammalian P2Y receptor or was a species homologue of the mammalian P2Y6 receptor, we screened two different human genomic libraries and a Southern blot with a p2y3 receptor probe, under low-stringency conditions that allowed the clear identification of the human P2Y6 receptor gene. Our data indicated that the human genome does not contain a receptor that is more homologous to the avian p2y3 receptor than the P2Y6 receptor. Taken together, these data further define the pharmacological selectivities of these UDP selective receptors and strongly suggest that the avian p2y3 receptor is a species homologue of the mammalian P2Y6 receptor. PMID- 9730914 TI - The 38-amino-acid form of pituitary adenylate cyclase-activating polypeptide induces neurite outgrowth in PC12 cells that is dependent on protein kinase C and extracellular signal-regulated kinase but not on protein kinase A, nerve growth factor receptor tyrosine kinase, p21(ras) G protein, and pp60(c-src) cytoplasmic tyrosine kinase. AB - The 38-amino-acid isoform of pituitary adenylate cyclase-activating polypeptide (PACAP38) elicits a robust outgrowth of neurites in cultured PC12 cells. Initiation of neurite outgrowth occurs within 4-8 hr after the addition of PACAP38. Treatment with PACAP38 does not elicit collateral activation of p140(trk) nerve growth factor receptor tyrosine kinase activity, nor is it associated with tyrosine phosphorylation of suc1-associated neurotrophic factor target, a selective target of neurotrophin tyrosine kinase receptors. Coadministration of epidermal growth factor with PACAP38 elicits an enhanced response. Induction of neurites is also observed on the addition of PACAP38 to dominant negative Src and Ras PC12 cell variants. PACAP38 stimulates extracellular signal-regulated kinase (Erk) activity >10-fold within 5 min, and the effect is augmented by cotreatment with epidermal growth factor. Pretreatment with the cAMP-dependent protein kinase-selective inhibitor, H-89, is ineffective as an antagonist of PACAP38-induced neurite outgrowth, whereas down-regulation of protein kinase C (PKC) by phorbol ester or incubation with PKC-selective inhibitors GF109203X and calphostin C effectively blocks PACAP38-stimulated neurite formation. Stimulation of Erk activity is inhibited by incubation with PD90859, a pharmacological antagonist of the threonine/tyrosine dual-specificity Erk. Inhibition of ligand-stimulated Erk activation prevents PACAP38-induced neurite outgrowth. Collectively, these findings indicate that PACAP38-stimulated neuritogenesis requires PKC and Erk activation but is independent of cAMP dependent protein kinase, nerve growth factor receptor tyrosine kinase, p21(ras) G protein, and pp60(c-src) cytoplasmic tyrosine kinase. PMID- 9730916 TI - Alpha2C-adrenoceptor-overexpressing mice are impaired in executing nonspatial and spatial escape strategies. AB - Drugs acting via alpha2-adrenoceptors modulate cognitive functions mediated via frontostriatothalamic feedback loops. The alpha2C-adrenoceptor subtype is expressed in the basal ganglia, hippocampus, and neocortex, areas that are involved in memory and other cognitive functions. alpha2C-Overexpressing (OE) mice were impaired in spatial or nonspatial water maze (WM) tests, and alpha2 antagonist treatment fully reversed the WM escape defect in OE mice. However, alpha2C-overexpression did not influence open field and passive avoidance behaviors or cortical EEG arousal or the actions of alpha2 agonist or antagonist drugs on these functions. Our results suggest that alpha2C-adrenoceptors can modulate navigation to a hidden or visible escape platform, whereas many other actions of alpha2-adrenergic agents, such as sedation, are not mediated via alpha2C-adrenoceptors. Therefore, alpha2-agonists lacking alpha2C-AR affinity or alpha2C-AR subtype-selective alpha2 antagonists could modulate functioning of frontostriatothalamic feedback loops more effectively than the current subtype nonselective drugs. PMID- 9730915 TI - The anesthetic steroid (+)-3alpha-hydroxy-5alpha-androstane-17beta-carbonitrile blocks N-, Q-, and R-type, but not L- and P-type, high voltage-activated Ca2+ current in hippocampal and dorsal root ganglion neurons of the rat. AB - High voltage-activated (HVA) Ca2+ current (ICa) was recorded from neonatal rat hippocampal and adult rat dorsal root ganglion neurons. In both cell types, (+) 3alpha-hydroxy-5alpha-androstane-17beta-carbonitrile [(+)-ACN], a neuroactive steroid, had no effect on nifedipine- (L-type) or omega-agatoxin IVA- (P-type) sensitive ICa. Selective blockade of N-type current with omega-conotoxin GVIA and of Q-type current with omega-conotoxin MVIIC indicated that (+)-ACN inhibits both N- and Q-type current components in both cell types. Current persisting after blockade of all other current components (R-type) was also sensitive to (+)-ACN. Half-blockade of (+)-ACN-sensitive HVA current occurred in the range of 3-25 microM, with N-type current somewhat more sensitive than Q- or R-type. The (+) ACN enantiomer, (-)-ACN, and pregnanolone were somewhat less effective at inhibiting total HVA current than (+)-ACN, whereas several steroid analogs, including alfaxalone, were relatively ineffective at inhibiting total HVA current. Neither guanosine-5'-O-(2-thio)diphosphate nor guanosine-5'-O-(3 thio)triphosphate altered the ability of (+)-ACN to inhibit HVA current in dorsal root ganglion neurons, indicating that (+)-ACN acts directly on Ca2+ channels. The partial selectivity exhibited by (+)-ACN among different HVA current components suggests that manipulations of steroid analogues may be a useful strategy in the generation of more selective, more potent, small-molecular-weight HVA channel blockers. PMID- 9730917 TI - The 5-hydroxytryptamine6 receptor-selective radioligand [3H]Ro 63-0563 labels 5 hydroxytryptamine receptor binding sites in rat and porcine striatum. AB - Ro 63-0563 [4-amino-N-(2,6 bis-methylamino-pyridin-4-yl)-benzene sulfonamide] is a high affinity 5-hydroxytryptamine6 (HT6) receptor antagonist with more than 100 fold selectivity for the 5-HT6 receptor compared with 69 other receptors and binding sites. The present study describes the properties of [3H]Ro 63-0563, the first selective 5-HT6 receptor radioligand. Specific binding of [3H]Ro 63-0563 (nonspecific binding defined in the presence of 10 microM methiothepin) to recombinant rat and human 5-HT6 receptors was saturable, rapid, and reversible with equilibrium dissociation constants (Kd) of 6.8 nM and 4.96 nM, respectively. The pharmacological profile of the rat 5-HT6 receptor labeled with [3H]Ro 63-0563 (methiothepin > D-lysergic acid diethylamide > clozapine approximately Ro 63-0563 > lisuride > ergotamine approximately Ro 04-6790 > 5-HT > amitriptyline approximately metergoline approximately mianserin approximately ritanserin > methysergide > mesulergine) was similar to that obtained by using either [3H]D lysergic acide diethylamide or [3H]5-HT as radioligand. In equilibrium binding studies with rat striatal membranes, [3H]Ro 63-0563 labeled a single binding site with Kd and Bmax values of 11. 7 nM and 175 fmol/mg protein, respectively. In porcine striatal membranes, [3H]Ro 63-0563 also labeled a single binding site with Kd and Bmax values of 8.0 nM and 130 fmol/mg protein, respectively. The affinities of 14 5-HT6 receptor ligands at this binding site were similar to those found for the recombinant rat and human 5-HT6 receptor, which suggested the presence of 5-HT6 receptors in porcine striatum. PMID- 9730918 TI - A single amino-acid substitution in the EP2 prostaglandin receptor confers responsiveness to prostacyclin analogs. AB - A high degree of homology between the four Gs-coupled prostaglandin (PG) receptors [EP2, EP4, prostacyclin (IP), PGD2 (DP)] and the four Gq/Gi-coupled receptors [EP1, EP3, PGF2alpha (FP), thromboxane A2 (TP)] suggests that prostaglandin receptors evolved functionally from an ancestral EP receptor before the development of distinct binding epitopes. If so, ligand selectivity should be determined by a limited number of amino acids. EP2 receptor transmembrane domain residues that are similar to those in the EP4 receptor but differ from those in the IP receptor were mutated to the corresponding IP receptor residue. Activation of the mutant receptors by PGE2 (EP2 ligand), iloprost (stable prostacyclin analog), and PGE1 (EP2/IP ligand) was determined using a cAMP-dependent reporter gene assay. A Leu304-to-tyrosine substitution in the seventh transmembrane domain enhanced iloprost potency approximately 100-fold. A glycine substitution at Ser120 in the third transmembrane domain had no effect on drug potency but improved the response of the Tyr304 mutant. The potency of the natural prostaglandins PGF2alpha and PGD2 was not enhanced by the mutations. In contrast, the potency of all prostaglandins was reduced 10- to 100-fold when arginine 302, which is thought to be a counterion for the prostaglandin carboxylic acid, was mutated. Thus, a single amino acid change resulted in a selective gain of function for iloprost, which is consistent with the proposed phylogeny of the prostaglandin receptors. PMID- 9730919 TI - Pharmacological analysis of sterol delta8-delta7 isomerase proteins with [3H]ifenprodil. AB - Sterol Delta8-Delta7 isomerases (SIs) catalyze the shift of the double bond from C8-9 to C7-8 in the B-ring of sterols. Surprisingly, the isoenzymes in fungi (ERG2p) and vertebrates [emopamil binding protein (EBP)] are structurally completely unrelated, whereas the sigma1 receptor, a mammalian protein of unknown function, bears significant similarity with the yeast ERG2p. Here, we compare the drug binding properties of SIs and related proteins with [3H]ifenprodil as a common high affinity radioligand (Kd = 1.4-19 nM), demonstrating an intimate pharmacological relationship among ERG2p, sigma1 receptor, and EBP. This renders SIs a remarkable example for structurally diverse enzymes with similar pharmacological profiles and the propensity to bind drugs from different chemical groups with high affinity. We identified a variety of experimental drugs with nanomolar affinity for the human EBP (Ki = 0.5-14 nM) such as MDL28815, AY9944, triparanol, and U18666A. These compounds, as well as the fungicide tridemorph and the clinically used drugs tamoxifen, clomiphene, amiodarone, and opipramol, inhibit the in vitro activity of the recombinant human EBP (IC50 = 0.015-54 microM). The high affinity of the human EBP for 3H-tamoxifen (Kd = 3 +/- 2 nM) implies that the EBP carries the previously described microsomal antiestrogen binding site. Interactions of the EBP with structurally diverse lipophilic amines suggest that novel compounds of related structure should be counterscreened for inhibition of the enzyme to avoid interference with sterol Delta8-Delta7 isomerization. PMID- 9730920 TI - Comprehensive, comprehensible, distributed and intelligent databases: current status. AB - MOTIVATION: It is only a matter of time until a user will see not many but one integrated database of information for molecular biology. Is this true? Is it a good thing? Why will it happen? Where are we now? What developments are fostering and what developments are impeding progress towards this end? SUPPLEMENTARY INFORMATION: A list of WWW resources devoted to database issues in molecular biology is available at http://www.mips.biochem.mpg.de CONTACT: frishman@mips.biochem.mpg.de PMID- 9730921 TI - The LabBase system for data management in large scale biology research laboratories. AB - MOTIVATION: The development of laboratory information management systems (LIMSs) for large scale biology research projects can be a challenging problem. Many such projects generate complex datasets via complex procedures that undergo continuous refinement. A key software challenge is to simplify the database-development task so that databases can be built and modified quickly enough to keep pace with changing project-requirements. RESULTS: LabBase extends the facilities offered by relational database systems to simplify the task of creating databases for large scale biology research projects. LabBase provides a structural object data model, similar to ACEDB, and adds to this the concepts of Materials, Steps, and States: Materials are objects representing the identifiable things that participate in a laboratory protocol; Steps are objects reporting the results of a laboratory or analytical procedure; and States are objects denoting places in a laboratory protocol. The system provides a data definition language for succinctly defining laboratory databases, and operations for conveniently storing and retrieving data in such databases. The system also provides support for workflow management. LabBase is implemented in Perl5 and provides a natural interface for laboratory application programs written in Perl. AVAILABILITY: The software is freely available. Contact the authors. CONTACT: nat@jax.org PMID- 9730922 TI - A model system for studying the integration of molecular biology databases. AB - MOTIVATION: Integration of molecular biology databases remains limited in practice despite its practical importance and considerable research effort. The complexity of the problem is such that an experimental approach is mandatory, yet this very complexity makes it hard to design definitive experiments. This dilemma is common in science, and one tried-and-true strategy is to work with model systems. We propose a model system for this problem, namely a database of genes integrating diverse data across organisms, and describe an experiment using this model. RESULTS: We attempted to construct a database of human and mouse genes integrating data from GenBank and the human and mouse genome-databases. We discovered numerous errors in these well-respected databases: approximately 15% of genes are apparently missing from the genome-databases; links between the sequence and genome-databases are missing for another 5-10% of the cases; about a third of likely homology links are missing between the genome-databases; 10-20% of entries classified as 'genes' are apparently misclassified. By using a model system, we were able to study the problems caused by anomalous data without having to face all the hard problems of database integration. CONTACT: nat@jax.org PMID- 9730923 TI - Issues in developing integrated genomic databases and application to the human X chromosome. AB - MOTIVATION: In the past decade, a vast amount of mapping data has been generated on the human X chromosome, without a mechanism which would provide a global view of exactly what has been achieved. Large datasets are available electronically, but in heterogeneous formats and with incompatible access modes. In addition, relationships between objects in different datasets are often not specified. RESULTS: We discuss the problem of integrating these data into one database and define a number of requirements that are vital for any integration approach. We have developed IXDB, the Integrated X chromosome database, which fulfils those requirements and aims at providing a global view on genomic data at a chromosomal level. IXDB represents a conceptual framework based on identifying, storing and analysing relationships between biological objects, and includes a series of tools to automate the integration of such information. It currently focuses on physical mapping data, as a starting point towards a map of the human X chromosome that should provide a uniform and global research resource for ongoing and future sequencing and functional studies. AVAILABILITY: IXDB is available at http://ixdb.mpimg-berlin-dahlem.mpg.de. The iace2ixdb software and a description of the Iace data format are available from the authors. CONTACT: hrc@genoscope.cns.fr PMID- 9730924 TI - LIGAND: chemical database for enzyme reactions. AB - MOTIVATION: The existing molecular biology databases focus on the sequence and structural aspects of biological macromolecules, i.e. DNAs, RNAs and proteins. However, in order to understand the functional aspects, it is essential to computerize the interaction of these molecules. Furthermore, living cells contain additional molecules, such as metabolic compounds and metal ions, that may also be considered as parts of the basic building blocks of life, but are not well organized in public databases. LIGAND chemical database is our attempt to solve these problems, at least for enzymatic reactions. RESULTS: LIGAND consists of two sections: ENZYME and COMPOUND. The ENZYME section is an extension of previous studies (Suyama et al. , Comput. Applic. Biosci., 9, 9-15, 1993), and it is a flat-file representation of 3303 enzymes and 2976 enzymatic reactions in the chemical equation format that can be parsed by machine. The COMPOUND section has been newly constructed for information on the nomenclature and chemical structures of compounds. It contains 5383 chemical compounds. Both ENZYME and COMPOUND entries contain rich cross-reference information, most of which is automatically generated by the DBGET/LinkDB system, thus providing the linkage between chemical and biological databases. LIGAND is updated daily, tightly coupled with the KEGG metabolic pathway database, and forms the basis for reconstruction and computation of pathways. AVAILABILITY: LIGAND can be accessed through the DBGET/LinkDB and KEGG systems in the Japanese GenomeNet database service via http://www.genome.ad.jp/. The flat-file format of the LIGAND database can be downloaded by anonymous FTP via ftp://kegg. genome.adjp/molecules/ligand/. CONTACT: goto@kuicr.kyoto-u.ac.jp; nishioka@scl.kyoto-u.ac.jp; kanehisa@kuicr.kyoto-u.ac.jp PMID- 9730925 TI - Automatic extraction of keywords from scientific text: application to the knowledge domain of protein families. AB - MOTIVATION: Annotation of the biological function of different protein sequences is a time-consuming process currently performed by human experts. Genome analysis tools encounter great difficulty in performing this task. Database curators, developers of genome analysis tools and biologists in general could benefit from access to tools able to suggest functional annotations and facilitate access to functional information. APPROACH: We present here the first prototype of a system for the automatic annotation of protein function. The system is triggered by collections of s related to a given protein, and it is able to extract biological information directly from scientific literature, i.e. MEDLINE abstracts. Relevant keywords are selected by their relative accumulation in comparison with a domain specific background distribution. Simultaneously, the most representative sentences and MEDLINE abstracts are selected and presented to the end-user. Evolutionary information is considered as a predominant characteristic in the domain of protein function. Our system consequently extracts domain-specific information from the analysis of a set of protein families. RESULTS: The system has been tested with different protein families, of which three examples are discussed in detail here: 'ataxia-telangiectasia associated protein', 'ran GTPase' and 'carbonic anhydrase'. We found generally good correlation between the amount of information provided to the system and the quality of the annotations. Finally, the current limitations and future developments of the system are discussed. AVAILABILITY: The current system can be considered as a prototype system. As such, it can be accessed as a server at http://columba.ebi.ac. uk:8765/andrade/abx. The system accepts text related to the protein or proteins to be evaluated (optimally, the result of a MEDLINE search by keyword) and the results are returned in the form of Web pages for keywords, sentences and s. SUPPLEMENTARY INFORMATION: Web pages containing full information on the examples mentioned in the text are available at: http://www.cnb.uam.es/ approximately cnbprot/keywords/ CONTACT: valencia@cnb.uam.es PMID- 9730926 TI - The effect of evenly spaced constant sites on the distribution of the random division of a molecular sequence. AB - MOTIVATION: A modified Sherman statistic can be used to test whether the differences between two aligned sequences are distributed at random along the sequences, or whether they are clustered, which suggests anomalies of evolution such as partial gene recombination or functional constraints. The presence of evenly spaced constant sites (such as constancy at the second codon position in genes coding for proteins) lowers the statistic and makes the significance less than it should be. RESULTS: The magnitude of the constant-site effect is shown by simulation to depend mainly on the proportion of differences between two sequences and on the number of constant sites that are added after each variable site. This latter number can be estimated from the variance of sites in a sequence matrix at the first, second and third codon positions, to obtain a ratio that corrects the statistic. When expressed as standard errors, the uncorrected results are too low (typically half to one unit when almost all the variation is at the third codon position). Correction raises the standard errors to levels close to expectation. If the data show no marked ternary periodicity, the correction is very small. The method is illustrated with biological data that show close to random behaviour, and with data that exhibit strong clustering. AVAILABILITY: The software is available from the author and has also been placed on the EMBL file server (Software@embl-ebi.ac.uk). CONTACT: phas1@le.ac.uk PMID- 9730927 TI - JOY: protein sequence-structure representation and analysis. AB - MOTIVATION: JOY is a program to annotate protein sequence alignments with three dimensional (3D) structural features. It was developed to display 3D structural information in a sequence alignment and to help understand the conservation of amino acids in their specific local environments. RESULTS: : The JOY representation now constitutes an essential part of the two databases of protein structure alignments: HOMSTRAD (http://www-cryst.bioc.cam.ac.uk/homstrad ) and CAMPASS (http://www-cryst.bioc.cam.ac. uk/campass). It has also been successfully used for identifying distant evolutionary relationships. AVAILABILITY: The program can be obtained via anonymous ftp from torsa.bioc.cam.ac.uk from the directory /pub/joy/. The address for the JOY server is http://www cryst.bioc.cam.ac.uk/cgi-bin/joy.cgi. CONTACT: kenji@cryst.bioc.cam.ac.uk PMID- 9730928 TI - An NMR assignment module implemented in the Gifa NMR processing program. AB - MOTIVATION: Peptide and protein structures are determined daily using NMR spectroscopy. Assignment of the NMR spectra is an important step within the procedure and is usually the limiting one. Computer-aided assignment tools should be user friendly with open architecture to communicate with other programs involved in the structure determination. RESULTS: Here we present an interactive NMR assignment module which provides numerous graphic tools for the user. The module is composed of a database management system-handling peaks, spins and spin systems. The assignment information is maintained as a set of interrelated associative arrays, which serve as generic high-level data structures. The module is developed in the macro language embedded in the Gifa NMR processing program (Pons et al. , J. Biomol. NMR, 8 , 445-452, 1996). This provides the user with a consistent interface, a set of sophisticated tools, and an easily extendible and customizable environment. AVAILABILITY: The program is available on request from the authors. The Gifa package can be accessed at: ((http://www.cbs. univ montp1.fr/GIFA)) CONTACT: Marc-Andre.Delsuc@cbs.univ-montp1.fr PMID- 9730929 TI - QU-GENE: a simulation platform for quantitative analysis of genetic models. AB - MOTIVATION: Classical quantitative genetics theory makes a number of simplifying assumptions in order to develop mathematical expressions that describe the mean and variation (genetic and phenotypic) within and among populations, and to predict how these are expected to change under the influence of external forces. These assumptions are often necessary to render the development of many aspects of the theory mathematically tractable. The availability of high-speed computers today provides opportunity for the use of computer simulation methodology to investigate the implications of relaxing many of the assumptions that are commonly made. RESULTS: QU-GENE (QUantitative-GENEtics) was developed as a flexible computer simulation platform for the quantitative analysis of genetic models. Three features of the QU-GENE software that contribute to its flexibility are (i) the core E(N:K) genetic model, where E is the number of types of environment, N is the number of genes, K indicates the level of epistasis and the parentheses indicate that different N:K genetic models can be nested within types of environments, (ii) the use of a two-stage architecture that separates the definition of the genetic model and genotype-environment system from the detail of the individual simulation experiments and (iii) the use of a series of interactive graphical windows that monitor the progress of the simulation experiments. The E(N:K) framework enables the generation of families of genetic models that incorporate the effects of genotype-by-environment (G x E) interactions and epistasis. By the design of appropriate application modules, many different simulation experiments can be conducted for any genotype environment system. The structure of the QU-GENE simulation software is explained and demonstrated by way of two examples. The first concentrates on some aspects of the influence of G x E interactions on response to selection in plant breeding, and the second considers the influence of multiple-peak epistasis on the evolution of a four-gene epistatic network. AVAILABILITY: QU-GENE is available over the Internet at (http://pig.ag.uq.edu.au/qu-gene/) CONTACT: m.cooper@mailbox.uq.edu. au PMID- 9730930 TI - Overview of HIV and AIDS: biology and epidemiology of the virus. AB - HIV-1 infects mononuclear cells using the CD4+ molecule and the chemical receptors of those cells. After a prolonged clinical latency period, the ability to replace destroyed cells is outpaced by ongoing cellular destruction, leading to the characteristic immunodeficiency of AIDS and its opportunistic infections and neoplasms. In the United States, the number of new cases of AIDS has diminished in recent years, although in some groups, such as women, the number of new cases continues to rise. In the developing world, AIDS remains a pandemic of huge proportions. In the absence of an effective vaccine, culturally appropriate efforts at education and behavior modification offer the best hope of controlling AIDS. PMID- 9730931 TI - Risk to the health care worker of HIV infection and how to minimize It. AB - Occupational transmission of HIV fortunately is uncommon. The risk of acquiring HIV depends on the mode of exposure, the body fluid involved, and the source patient. Percutaneous injuries carry the greatest risk (approximately 0.3%), and blood is by far the most important source of HIV to which the health care worker is exposed. Universal precautions should be applied to all patients in order to decrease the risk of occupational transmission of HIV. Furthermore, a system must be designed to provide adequate assessment, counselling, and follow-up for exposed health care personnel. Postexposure prophylaxis must be tailored to the specific exposure for each health care worker. PMID- 9730932 TI - Oropharyngeal lesions in AIDS. AB - Any physician who is treating HIV-infected patients can expect to deal with individuals who are or will be experiencing the physical manifestations of the disease. Fortunately, most of these conditions can be successfully treated; other cases have palliative treatments that can improve quality of life and assist in pain reduction and suffering. All patients with HIV need to be managed jointly throughout their illnesses so that these patients do not receive conflicting advice, prescriptions, and so forth. Also, patients benefit from receiving care from the specialist that is most appropriate for their condition. The physician or dentist working alone can often help the patient; when they work together, however, optimum care is likely the result. PMID- 9730933 TI - Esophageal disease in patients with AIDS: diagnosis and treatment. AB - Patients with HIV infection often present with symptoms suggesting esophageal disease: these include odynophagia (pain with swallowing), dysphagia (difficulty in swallowing), and retrosternal chest pain. Esophageal symptoms rank second only to diarrhea in frequency of gastrointestinal complaints among patients with AIDS. Also, esophageal opportunistic infections have been associated with a poor outcome, the mean survival after diagnosis being less than 6 months in one study. Such short survival may be explained by the underlying immunosuppression, as well as a decrease in nutritional intake due to difficulty swallowing. PMID- 9730934 TI - Gastric disorders and nutritional management. AB - Protein calorie malnutrition is a frequent complication of HIV and AIDS. Malnutrition has been one of the most common AIDS-defining conditions recognized by the Centers for Disease Control and Prevention. Proper nutritional management can have a positive impact on the clinical course of HIV-infected patients. Further information is required in order to assign nutrition its proper priority in the clinical management of HIV-infected individuals. The use of gastrostomy feedings in patients with AIDS is similar to the use of these feedings in other diseases. PMID- 9730935 TI - Correlation of intestinal structure and function in HIV infection. AB - The gastrointestinal tract has great importance in HIV infection because of its role as a primary barrier to the external environment and consequent need for effective immune function. Many factors promote the development of diarrhea in HIV-infected individuals. Understanding the genesis of the symptom is key to formulating effective therapy. Ultimate control of the problem depends on preventing HIV replication and immune depletion, as well as avoiding the development of opportunistic enteric infections in patients with severe immune deficiency. PMID- 9730936 TI - Evaluation of diarrhea in HIV-infected patients. AB - Diarrhea is a major problem for patients infected with HIV: initial studies indicated that 50% of HIV-seropositive patients developed diarrhea, but this may be an underestimate. Diarrhea has an appreciable adverse affect on the quality of life of these patients; also, they use more health care facilities and health care dollars than HIV-positive patients without diarrhea. Individuals who have homosexuality or bisexuality as their HIV risk factor are more likely to have diarrhea and to have an enteric pathogen identified as the cause of diarrhea than are patients who have heterosexuality or intravenous drug use as their risk factor. PMID- 9730937 TI - Small intestine pathogens in AIDS: conventional and opportunistic. AB - The small intestine, coming in direct contact with ingested potential pathogens, depends on active mucosal immunity to withstand invasion and damage. In patients with AIDS and severe impairment of immunoregulatory lymphocytes, proliferation of protozoal, viral, bacterial, and fungal pathogens produces diarrhea and malabsorption. When noninvasive tests of stool and blood fail to identify responsible organisms, endoscopy can reveal mucosal lesions which are suggestive if not diagnostic. Cryptosporidium, cf2E. intestinalis, cf1CMV, MAC, and other infections can be identified by intestinal biopsy quicker and often at lower overall cost than they can be by culture. PMID- 9730938 TI - Diagnosis and treatment of colonic disease in AIDS. AB - The colon is a frequent site of gastrointestinal complications in patients with HIV infection, and these colonic disorders increase in frequency as immunodeficiency worsens. The most common clinical manifestations of colonic disease in AIDS are diarrhea, lower gastrointestinal bleeding, and abdominal pain. Toxic megacolon, intussuseption, typhlitis, idiopathic colonic ulcer, and pneumatosis intestinalis also have been described. In the HIV-infected patient with preserved immunity, the most common cause of colitis is bacterial, but as the degree of immunodeficiency worsens, opportunistic pathogens (CMV, protozoa, mycobacteria, fungi) and neoplasms become more frequent. The frequent use of antibiotics, chemotherapeutic agents, and frequent hospitalization increase the susceptibility to cf2Clostridium difficule cf1colitis. Endoscopy plays an integral role in the management of many colonic disorders in AIDS. PMID- 9730939 TI - Anorectal pathology in AIDS. AB - Anorectal complaints are common in persons with AIDS and are being seen increasingly because advances in therapy for HIV, such as the new antiretroviral protease inhibitors, have resulted in longer life expectancy for those with HIV infection. In the past, many patients with HIV infection were seen at referral centers; now, however, primary care physicians as well as gastroenterologists and surgeons in the community are managing and caring for these patients. For this reason, it is important for clinicians to recognize the spectrum of anorectal disease in patients with AIDS, as well as its appropriate evaluation and management. PMID- 9730940 TI - Gastrointestinal bleeding in AIDS. AB - Gastrointestinal (GI) bleeding is an uncommon manifestation of AIDS despite the frequent involvement of the GI tract by infections, malignancies, and a variety of other disorders. GI bleeding may occur from a variety of causes, some specifically associated with AIDS and others unrelated to immunocompromise; most patients with AIDS who have lower GI bleeding have causes attributable to AIDS specific lesions. Just as in an incompetent patient, an aggressive approach to diagnosis and treatment of GI bleeding in AIDS patients is warranted. PMID- 9730941 TI - HIV-Related hepatic disease: when and why to biopsy. AB - Patients infected with HIV are susceptible to a variety of hepatic processes that are related to immunosuppression or are associated with the risk factors of homosexuality and parenteral drug use. These processes present in a myriad of ways including fever, right upper quadrant pain, and hepatomegaly, or simply as asymptomatic elevations of liver tests. Care of these patients demands systematic evaluation and treatment to ensure that morbidity and mortality are minimized and quality of life and medical care costs are optimized. PMID- 9730942 TI - AIDS-Related biliary tract disease. AB - Increasingly, specialists caring for patients with AIDS have reported relatively small numbers of patients with biliary tract disease. These conditions fall into three general categories: (1) non-HIV-associated conditions of the bile duct, (2) acalculous cholecystitis, and (3) AIDS cholangiopathy. Whatever the cause, a sizable percentage of patients with AIDS are found to have abnormal biliary tract morphology. It is essential for the clinician first to exclude biliary tract conditions such as gallstone disease and then to clearly investigate those patients with clinical, biochemical, or radiographic features suggestive of papillary stenosis. Patients with AIDS-associated papillary stenosis do respond symptomatically to ERCP sphincterotomy. PMID- 9730943 TI - Laparoscopy in AIDS. AB - Surgical problems in the patient with HIV and AIDS are becoming more commonplace as the incidence of HIV seropositivity increases and patients with AIDS are living longer. Laparoscopic surgery is being applied more frequently in the diagnosis and therapy of these patients' problems as more surgeons become familiar with the techniques. Although no prospectively randomized trials exist, the benefits of the laparoscopic approach clearly have had an impact on the morbidity associated with surgery. The decreased perioperative immune depression may benefit these patients, and risks to the operative teams probably are lessened if basic tenets of laparoscopic surgery are observed. PMID- 9730944 TI - The mechanism of IL-5 signal transduction. AB - Cytokines are important regulators of hematopoiesis. They exert their actions by binding to specific receptors on the cell surface. Interleukin-5 (IL-5) is a critical cytokine that regulates the growth, activation, and survival of eosinophils. Because eosinophils play a seminal role in the pathogenesis of asthma and allergic diseases, an understanding of the signal transduction mechanism of IL-5 is of paramount importance. The IL-5 receptor is a heterodimer of alpha- and beta-subunits. The alpha-subunit is specific, whereas the beta subunit is common to IL-3, IL-5, and granulocyte/macrophage colony-stimulating factor (GM-CSF) receptors and is crucial for signal transduction. It has been shown that there are two major signaling pathways of IL-5 in eosinophils. IL-5 activates Lyn, Syk, and JAK2 and propagates signals through the Ras-MAPK and JAK STAT pathways. Studies suggest that Lyn, Syk, and JAK2 tyrosine kinases and SHP-2 tyrosine phosphatase are important for eosinophil survival. In contrast to their survival-promoting activity, Lyn and JAK2 appear to have no role in eosinophil degranulation or expression of surface adhesion molecules. Raf-1 kinase, on the other hand, is critical for eosinophil degranulation and adhesion molecule expression. Btk is involved in IL-5 stimulation of B cell function. However, it does not appear to be important for eosinophil function. Thus a clear segregation of signaling molecules based on their functional importance is emerging. This review describes the signal transduction mechanism of the IL-3/GM-CSF/IL-5 receptor system and compares and contrasts IL-5 signaling between eosinophils and B cells. PMID- 9730945 TI - Stepping back and looking forward: downregulation of G proteins as a mechanism of desensitization in tissues. Focus on "Carbachol-induced desensitization of PLC beta pathway in rat myometrium: downregulation of Gqalpha/G11alpha". PMID- 9730946 TI - Carbachol-induced desensitization of PLC-beta pathway in rat myometrium: downregulation of Gqalpha/G11alpha. AB - In the estrogen-treated rat myometrium, carbachol increased the generation of inositol phosphates by stimulating the muscarinic receptor-Gq/G11-phospholipase C beta3 (PLC-beta3) cascade. Exposure to carbachol resulted in a rapid and specific (homologous) attenuation of the subsequent muscarinic responses in terms of inositol phosphate production, PLC-beta3 translocation to membrane, and contraction. Refractoriness was accompanied by a reduction of membrane muscarinic binding sites and an uncoupled state of residual receptors. Protein kinase C (PKC) altered the functionality of muscarinic receptors and contributed to the initial period of desensitization. A delayed phase of the muscarinic refractoriness was PKC independent and was associated with a downregulation of Gqalpha/G11alpha. Atropine failed to induce desensitization as well as Gqalpha/G11alpha downregulation, indicating that both events involve active occupancy of the receptor. Prolonged exposure to AlF-4 reduced subsequent AlF-4 as well as carbachol-mediated inositol phosphate responses and similarly induced downregulation of Gqalpha/G11alpha. Data suggest that a decrease in the level of Gqalpha/G11alpha is subsequent to its activation and may account for heterologous desensitization. PMID- 9730947 TI - Voltage-dependent block of endothelial volume-regulated anion channels by calix[4]arenes. AB - We have studied the effects of calix[4]arenes on the volume-regulated anion channel (VRAC) currents in cultured calf pulmonary artery endothelial cells. TS- and TS-TM-calix[4]arenes induced a fast inhibition at positive potentials but were ineffective at negative potentials. Maximal block occurred at potentials between 30 and 50 mV. Lowering extracellular pH enhanced the block and shifted the maximum inhibition to more negative potentials. Current inhibition was also accompanied by an increased current noise. From the analysis of the calix[4]arene induced noise, we obtained a single-channel conductance of 9.3 +/- 2.1 pS (n = 9) at +30 mV. The voltage- and time-dependent block were described using a model in which calix[4]arenes bind to a site at an electrical distance of 0.25 inside the channel with an affinity of 220 microM at 0 mV. Binding occludes VRAC at moderately positive potentials, but calix[4]arenes permeate the channel at more positive potentials. In conclusion, our data suggest an open-channel block of VRAC by calix[4]arenes that also depends on the protonation of the binding site within the pore. PMID- 9730948 TI - Prostaglandin F2alpha stimulates CFTR activity by PKA- and PKC-dependent phosphorylation. AB - The cystic fibrosis transmembrane conductance regulator (CFTR) can be activated by protein kinase A (PKA)- or protein kinase C (PKC)-dependent phosphorylation. To understand how activation of both kinases affects CFTR activity, transfected NIH/3T3 cells were stimulated with forskolin (FSK), phorbol myristate acetate (PMA), or prostaglandin F2alpha (PGF). PGF stimulates inositol trisphosphate and cAMP production in NIH/3T3 cells. As measured by I- efflux, maximal CFTR activity with PGF and FSK was equivalent and fivefold greater than that with PMA. Both PGF and PMA had additive effects on FSK-dependent CFTR activity. PMA did not increase cellular cAMP, and maximal PGF-dependent CFTR activity occurred with approximately 20% of the cellular cAMP observed with FSK-dependent activation. Staurosporine, but not H-89, inhibited CFTR activation and in vivo phosphorylation at low PGF concentrations. In contrast, at high PGF concentrations, CFTR activation and in vivo phosphorylation were inhibited by H 89. As judged by protease digestion, the sites of in vivo CFTR phosphorylation with FSK and PMA differed. For PGF, the data were most consistent with in vivo CFTR phosphorylation by PKA and PKC. Our data suggest that activation of PKC can enhance PKA-dependent CFTR activation. PMID- 9730949 TI - Age-associated increase in PGE2 synthesis and COX activity in murine macrophages is reversed by vitamin E. AB - We previously showed that increased macrophage and PGE2 production with age is due to enhanced cyclooxygenase (COX) activity and COX-2 expression. This study determined the effect of vitamin E supplementation on macrophage PGE2 synthesis in young and old mice and its underlying mechanism. Mice were fed 30 or 500 parts per million vitamin E for 30 days. Lipopolysaccharide (LPS)-stimulated macrophages from old mice produced significantly more PGE2 than those from young mice. Vitamin E supplementation reversed the increased PGE2 production in old mice but had no effect on macrophage PGE2 production in young mice. In both LPS stimulated and unstimulated macrophages, COX activity was significantly higher in old than in young mice at all intervals. Vitamin E supplementation completely reversed the increased COX activity in old mice to levels comparable to those of young mice but had no effect on macrophage COX activity of young mice or on COX-1 and COX-2 protein or COX-2 mRNA expression in young or old mice. Thus vitamin E reverses the age-associated increase in macrophage PGE2 production and COX activity. Vitamin E exerts its effect posttranslationally, by inhibiting COX activity. PMID- 9730950 TI - Functional expression of putative H+-K+-ATPase from guinea pig distal colon. AB - A guinea pig cDNA encoding the putative colonic H+-K+-ATPase alpha-subunit (T. Watanabe, M. Sato, K. Kaneko, T. Suzuki, T. Yoshida, and Y. Suzuki; GenBank accession no. D21854) was functionally expressed in HEK-293, a human kidney cell line. The cDNA for the putative colonic H+-K+-ATPase was cotransfected with cDNA for either rabbit gastric H+-K+-ATPase or Torpedo Na+-K+-ATPase beta-subunit. In both expressions, Na+-independent, K+-dependent ATPase (K+-ATPase) activity was detected in the membrane fraction of the cells, with a Michaelis-Menten constant for K+ of 0.68 mM. The expressed K+-ATPase activity was inhibited by ouabain, with its IC50 value being 52 microM. However, the activity was resistant to Sch 28080, an inhibitor specific for gastric H+-K+-ATPase. The ATPase was not functionally expressed in the absence of the beta-subunits. Therefore, it is concluded that the cDNA encodes the catalytic subunit (alpha-subunit) of the colonic H+-K+-ATPase. Although the beta-subunit of the colonic H+-K+-ATPase has not been identified yet, both gastric H+-K+-ATPase and Na+-K+-ATPase beta subunits were found to act as a surrogate for the colonic beta-subunit for the functional expression of the ATPase. The present colonic H+-K+-ATPase first expressed in mammalian cells showed the highest ouabain sensitivity in expressed colonic H+-K+-ATPases so far reported (rat colonic in Xenopus oocytes had an IC50 = 0.4-1 mM; rat colonic in Sf9 cells had no ouabain sensitivity). PMID- 9730951 TI - GLP-1 action in L6 myotubes is via a receptor different from the pancreatic GLP-1 receptor. AB - The incretin hormone glucagon-like peptide-1 (GLP-1)-(7-36) amide is best known for its antidiabetogenic actions mediated via a GLP-1 receptor present on pancreatic endocrine cells. To investigate the molecular mechanisms of GLP-1 action in muscle, we used cultured L6 myotubes. In L6 myotubes, GLP-1 enhanced insulin-stimulated glycogen synthesis by 140% while stimulating CO2 production and lactate formation by 150%. In the presence of IBMX, GLP-1 diminished cAMP levels to 83% of IBMX alone. In L6 myotubes transfected with pancreatic GLP-1 receptor, GLP-1 increased cAMP levels and inhibited glycogen synthesis by 60%. An antagonist of pancreatic GLP-1 receptor, exendin-4-(9-39), inhibited GLP-1 mediated glycogen synthesis in GLP-1 receptor-transfected L6 myotubes. However, in parental L6 myotubes, exendin-4-(9-39) and GLP-1-(1-36) amide, an inactive peptide on pancreatic GLP-1 receptor, displaced 125I-labeled GLP-1 binding and stimulated glycogen synthesis by 186 and 130%, respectively. These results suggest that the insulinomimetic effects of GLP-1 in L6 cells are likely to be mediated by a receptor that is different from the GLP-1 receptor found in the pancreas. PMID- 9730952 TI - Differences in contractile protein content and isoforms in phasic and tonic smooth muscles. AB - The basis of tonic vs. phasic contractile phenotypes of visceral smooth muscles is poorly understood. We used gel electrophoresis and quantitative scanning densitometry to measure the content and isoform composition of contractile proteins in opossum lower esophageal sphincter (LES), to represent tonic muscle, and circular muscle of the esophageal body (EB), to represent phasic smooth muscle. The amount of protein in these two types of muscles is similar: approximately 27 mg/g of frozen tissue. There is no difference in the relative proportion of myosin, actin, calponin, and tropomyosin in the two muscle types. However, the EB contains approximately 2.4-times more caldesmon than the LES. The relative ratios of alpha- to gamma-contractile isoforms of actin are 0.9 in the LES and 0.3 in EB. The ratio between acidic (LC17a) and basic (LC17b) isoforms of the 17-kDa essential light chain of myosin is 0.7:1 in the LES, compared with 2.7:1 in the EB. There is no significant difference in the ratios of smooth muscle myosin SM1 and SM2 isoforms in the two muscle types. The level of the myosin heavy chain isoform, which contains the seven-amino acid insert in the myosin head, is about threefold higher in the EB compared with LES. In conclusion, the esophageal phasic muscle in contrast to the tonic LES contains proportionally more caldesmon, LC17a, and seven-amino acid-inserted myosin and proportionally less alpha-actin. These differences may provide a basis for functional differences between tonic and phasic smooth muscles. PMID- 9730953 TI - Regulation of apical membrane Na+/H+ exchangers NHE2 and NHE3 in intestinal epithelial cell line C2/bbe. AB - We examined the regulation of the Na+/H+ exchangers (NHEs) NHE2 and NHE3 by expressing them in human intestinal C2/bbe cells, which spontaneously differentiate and have little basal apical NHE activity. Unidirectional apical membrane 22Na+ influxes were measured in NHE2-transfected (C2N2) and NHE3 transfected (C2N3) cells under basal and stimulated conditions, and their activities were distinguished as the HOE-642-sensitive and -insensitive components of 5-(N,N-dimethyl)amiloride-inhibitable flux. Both C2N2 and C2N3 cells exhibited increased apical membrane NHE activity under non-acid-loaded conditions compared with nontransfected control cells. NHE2 was inhibited by 8-(4 chlorophenylthio)adenosine 3',5'-cyclic monophosphate and thapsigargin, was stimulated by serum, and was unaffected by cGMP- and protein kinase C-dependent pathways. In contrast, NHE3 was inhibited by all regulatory pathways examined. Under acid-loaded conditions (which increase apical Na+ influx), NHE2 and NHE3 exhibited similar patterns of regulation, suggesting that the second messenger effects observed were not secondary to effects on cell pH. Thus, in contrast to their expression in nonepithelial cells, NHE2 and NHE3 expressed in an epithelial cell line behave similarly to endogenously expressed intestinal apical membrane NHEs. We conclude that physiological regulation and function of epithelium specific NHEs are dependent on tissue-specific factors and/or conditional requirements. PMID- 9730954 TI - Alpha-adrenergic receptors regulate human lymphocyte amiloride-sensitive sodium channels. AB - Two independent signal transduction pathways regulate lymphocyte amiloride sensitive sodium channels (ASSCs), one utilizing cAMP as a second messenger and the other utilizing a GTP-binding protein. This implies that two plasma membrane receptors play a role in the regulation of lymphocyte ASSCs. In this study, we tested the hypothesis that alpha1- and alpha2-adrenergic receptors independently regulate lymphocyte ASSCs via the two previously identified second messengers. Direct measurements indicated that norepinephrine increased lymphocyte cAMP and activated ASSCs. The alpha2-specific inhibitor, yohimbine, blocked this activation, thereby linking alpha2-adrenergic receptors to ASSC regulation via cAMP. The alpha1-specific ligand, terazosin, acted as an agonist and activated lymphocyte ASSCs but inhibited ASSC current that had been preactivated by norepinephrine or 8-(4-chlorophenylthio) (CPT)-cAMP. Terazosin had no effect on the lymphocyte whole cell ASSC currents preactivated by treatment with pertussis toxin. This finding indirectly links alpha1-adrenergic receptors to lymphocyte ASSC regulation via GTP-binding proteins. Terazosin had no direct inhibitory or stimulatory effects on alpha,beta,gamma-endothelial sodium channels reconstituted into planar lipid bilayers and expressed in Xenopus oocytes, ruling out a direct interaction between terazosin and the channels. These findings support the hypothesis that both alpha1- and alpha2-adrenergic receptors independently regulate lymphocyte ASSCs via GTP-binding proteins and cAMP, respectively. PMID- 9730955 TI - Polarity, integrin, and extracellular matrix dynamics in the postischemic rat kidney. AB - Acute renal failure (ARF) as a consequence of ischemic injury is a common disease affecting 5% of the hospitalized population. Despite the fact that mortality from ARF is high, there has been little improvement in survival rates over the last 40 years. The pathogenesis of ARF may be related to substantial changes in cell-cell and cell-extracellular matrix interactions mediated by beta1-integrins. On the basis of in vitro and in vivo studies, reorganization of beta1-integrins from basal to apical surfaces of injured tubular epithelia has been suggested to facilitate epithelial detachment, contributing to tubular obstruction and backleak of glomerular filtrate. In this study, we examine integrin and extracellular matrix dynamics during epithelial injury and repair using an in vivo rat model of unilateral ischemia. We find that, soon after reperfusion, beta1-integrins newly appear on lateral borders in epithelial cells of the S3 segment but are not on the apical surface. At later times, as further injury and regeneration coordinately occur, epithelia adherent to the basement membrane localize beta1 predominantly to basal surfaces even while the polarity of other marker proteins is lost. At the same time, amorphous material consisting of depolarized exfoliated cells fills the luminal space. Notably, beta1-integrins are not detected on exfoliated cells. A novel finding is the presence of fibronectin, a glycoprotein of plasma and the renal interstitium, in tubular spaces of the distal nephron and to a lesser extent S3 segments. These results indicate that beta1-integrins dramatically change their distribution during ischemic injury and epithelial repair, possibly contributing to cell exfoliation initially and to epithelial regeneration at later stages. Together with the appearance of large amounts of fibronectin in tubular lumens, these alterations may play a significant role in the pathophysiology of ARF. PMID- 9730956 TI - Optical measurement of stimulus-evoked membrane dynamics in single pancreatic acinar cells. AB - Stimulation of pancreatic acinar cells induces the release of digestive enzymes via the exocytotic fusion of zymogen granules and activates postfusion granule membrane retrieval and receptor cycling. In the present study, changes in membrane surface area of rat single pancreatic acinar cells were monitored by cell membrane capacitance (Cm) measurements and by the membrane fluorescent dye FM1-43. When measured with the Cm method, agonist treatment evoked a graded, transient increase in acinar cell surface area averaging 3. 5%. In contrast, a 13% increase in surface area was estimated using FM1-43, corresponding to the fusion of 48 zymogen granules at a rate of 0.5 s-1. After removal of FM1-43 from the surface-accessible membrane, a residual fluorescence signal was shown by confocal microscopy to be localized in endosome-like structures and confined to the apical regions of acinar cells. The development of an optical method for monitoring the membrane turnover of single acinar cells, in combination with measurements of Cm changes, reveals coincidence of exocytotic and endocytotic activity in acinar cells after hormonal stimulation. PMID- 9730957 TI - Induction of Mn SOD in human monocytes without inflammatory cytokine production by a mutant endotoxin. AB - Endotoxin selectively induces monocyte Mn superoxide dismutase (SOD) without affecting levels of Cu,Zn SOD, catalase, or glutathione peroxidase. However, little is known about the structure-activity relationship and the mechanism by which endotoxin induces Mn SOD. In this study we demonstrated that a mutant Escherichia coli endotoxin lacking myristoyl fatty acid at the 3' R-3 hydroxymyristate position of the lipid A moiety retained its full capacity to coagulate Limulus amoebocyte lysate compared with the wild-type E. coli endotoxin and markedly stimulated the activation of human monocyte nuclear factor-kappaB and the induction of Mn SOD mRNA and enzyme activity. However, in contrast to the wild-type endotoxin, it failed to induce significant production of tumor necrosis factor-alpha and macrophage inflammatory protein-1alpha by monocytes and did not induce the phosphorylation and nuclear translocation of mitogen-activated protein kinase. These results suggest that 1) lipid A myristoyl fatty acid, although it is important for the induction of inflammatory cytokine production by human monocytes, is not necessary for the induction of Mn SOD, 2) endotoxin-mediated induction of Mn SOD and inflammatory cytokines are regulated, at least in part, through different signal transduction pathways, and 3) failure of the mutant endotoxin to induce tumor necrosis factor-alpha production is, at least in part, due to its inability to activate mitogen-activated protein kinase. PMID- 9730958 TI - Conductive pathways for chloride and oxalate in rabbit ileal brush-border membrane vesicles. AB - To evaluate the possibility that an apical membrane conductive pathway for oxalate is present in the rabbit distal ileum, we studied oxalate ([14C]oxalate) and chloride (36Cl) uptake into brush-border membrane vesicles enriched 15- to 18 fold in sucrase activity. Voltage-sensitive pathways for oxalate and chloride were identified by the stimulation of uptake provided by an inwardly directed potassium diffusion potential in the presence of valinomycin. Additionally, outwardly directed oxalate (or chloride) gradients stimulated [14C]oxalate (or 36Cl) uptake to a greater degree in the absence of valinomycin (when intracellular and extracellular potassium are equal) than in the presence of valinomycin. Voltage-dependent anion uptake was poorly saturable: apparent affinity constants were 141 +/- 17 and 126 +/- 8 mM for chloride and oxalate, respectively. Activation energies for the voltage-dependent uptake processes were low: 4.7 and 6.3 kcal/mol for chloride and oxalate, respectively. Sensitivity profiles of voltage-dependent chloride and oxalate uptake to anion transport inhibitors were similar. We conclude that an anion conductance is present in the apical membranes of ileal enterocytes and that this conductance is a candidate pathway for oxalate efflux from the enterocyte during transepithelial oxalate secretion. PMID- 9730959 TI - Regulation by retinoids of P2Y2 nucleotide receptor mRNA in human uterine cervical cells. AB - Extracellular ATP stimulates acute changes in paracellular permeability across cultures of human uterine cervical epithelial cells [G. I. Gorodeski, D. E. Peterson, B. J. De Santis, and U. Hopfer. Am. J. Physiol. 270 (Cell Physiol. 39): C1715-C1725, 1996]. In this paper, we characterize mRNA for a P2Y2 nucleotide receptor in human cervical cells. Using oligonucleotide primers based on the sequence of human airway epithelium P2Y2 receptor, a single 632-bp cDNA band was identified in RT-PCR experiments in extracts of human endocervical and ectocervical tissues and in lysates of human cervical CaSki cells, but not in 3T3 fibroblasts. The nucleotide sequence was homologous to the corresponding human airway epithelium P2Y2 receptor. Northern blot analyses revealed hybridization of the P2Y2 receptor probe to a 2.0-kb mRNA fragment, as well as to 2.2-, 3. 0-, and 4.6-kb species, indicating that human cervical cells express P2Y2 receptor mRNA. Incubation of CaSki cells in retinoid-free medium abolished the ATP-induced changes in permeability and decreased the expression of the P2Y2 receptor mRNA; treatment with retinoids restored the responses to ATP and upregulated the P2Y2 receptor mRNA, suggesting that the receptor mediates ATP-related changes in permeability. Treatment with actinomycin D decreased the expression of the P2Y2 receptor RNA, but the ratio density of the receptor RNA relative to glyceraldehyde-3-phosphate dehydrogenase RNA remained unchanged, suggesting that retinoids upregulate transcription of the receptor mRNA. We conclude that retinoid-dependent modulation of the P2Y2 receptor expression, and hence of the responses to ATP, may be an important mechanism for the regulation of secretion of cervical mucus in vivo. PMID- 9730960 TI - Interstitial ATP level and degradation in control and postmyocardial infarcted rats. AB - With the aim of estimating interstitial levels and the breakdown process of ATP, cardiac microdialysis was performed in the left ventricular wall of in situ control and postinfarcted as well as of isolated, Langendorff-perfused rat hearts. With the use of a bioluminescence technique for dialysate ATP measurement, the baseline interstitial fluid ATP concentration in in situ hearts was estimated to be 38 +/- 8 nM. Regional ischemia induced an early peak increase in interstitial fluid ATP to 373 +/- 73 nM that correlates with the maximal incidence of ventricular arrhythmias. During continuous infusion of individual adenine nucleotides (50 microM ATP, ADP, or AMP), the dialysate samples were analyzed for adenine nucleotides, nucleosides, and bases using HPLC with ultraviolet detection. The patterns of catabolites were consistent with the major pathway of metabolism, that is, sequential dephosphorylation catalyzed by a chain of separate ecto-nucleotidases. In in situ postinfarcted hearts as well as in perfused hearts, a reduced catabolism rate of extracellular adenine nucleotides was observed. In conclusion, in in situ rat hearts, ATP can be released in substantial amounts in the interstitium where it readily undergoes enzymatic degradation. Dephosphorylation occurs sequentially and faster in in situ control hearts than in in situ postinfarcted or in perfused hearts. PMID- 9730961 TI - Stimulation of Na+-K+-2Cl- cotransporter in neuronal cells by excitatory neurotransmitter glutamate. AB - Na+-K+-2Cl- cotransporters are important in renal salt reabsorption and in salt secretion by epithelia. They are also essential in maintenance and regulation of ion gradients and cell volume in both epithelial and nonepithelial cells. Expression of Na+-K+-2Cl- cotransporters in brain tissues is high; however, little is known about their function and regulation in neurons. In this study, we examined regulation of the Na+-K+-2Cl- cotransporter by the excitatory neurotransmitter glutamate. The cotransporter activity in human neuroblastoma SH SY5Y cells was assessed by bumetanide-sensitive K+ influx, and protein expression was evaluated by Western blot analysis. Glutamate was found to induce a dose- and time-dependent stimulation of Na+-K+-2Cl- cotransporter activity in SH-SY5Y cells. Moreover, both the glutamate ionotropic receptor agonist N-methyl-D aspartic acid (NMDA) and the metabotropic receptor agonist (+/-)-1 aminocyclopentane-trans-1,3-dicarboxylic acid (trans-ACPD) significantly stimulated the cotransport activity in these cells. NMDA-mediated stimulation of the Na+-K+-2Cl- cotransporter was abolished by the selective NMDA-receptor antagonist (+)-MK-801 hydrogen maleate. trans-ACPD-mediated effect on the cotransporter was blocked by the metabotropic receptor antagonist (+)-alpha methyl-(4-carboxyphenyl)glycine. The results demonstrate that Na+-K+-2Cl- cotransporters in neurons are regulated by activation of both ionotropic and metabotropic glutamate receptors. PMID- 9730962 TI - Inducible expression of protein kinase Calpha suppresses steroidogenesis in Y-1 adrenocortical cells. AB - We have previously shown that protein kinase C (PKC) suppresses steroidogenesis in Y-1 adrenocortical cells. To ask directly if the PKCalpha isoform mediates this suppression, we have developed Y-1 cell lines in which PKCalpha is expressed from a tetracycline-regulated promoter. Induction of PKCalpha expression in these cell lines results in decreased P450 cholesterol side-chain cleavage enzyme (P450 SCC) activity as judged by the conversion of hydroxycholesterol to pregnenolone. Transcription of a P450-SCC promoter-luciferase construct is also reduced when PKCalpha expression is increased. However, expression of PKCalpha has no effect on 8-bromo-cAMP induction of steroidogenesis, indicating that these pathways function independently to regulate steroidogenesis. To determine the relationship between endogenous PKC activity and steroidogenesis, we examined 12 Y-1 subclones that were isolated by limited dilution cloning. In each of these subclones, steroid production correlates inversely with total PKC activity and with the expression of PKCalpha but not PKCepsilon or PKCzeta. These studies define for the first time the role of a specific PKC isoform (PKCalpha) in regulating steroidogenesis and P450-SCC activity in adrenocortical cells. PMID- 9730963 TI - PKA induces Ca2+ release and enhances ciliary beat frequency in a Ca2+-dependent and -independent manner. AB - The intent of this work was to evaluate the role of cAMP in regulation of ciliary activity in frog mucociliary epithelium and to examine the possibility of cross talk between the cAMP- and Ca2+-dependent pathways in that regulation. Forskolin and dibutyryl cAMP induced strong transient intracellular Ca2+ concentration ([Ca2+]i) elevation and strong ciliary beat frequency enhancement with prolonged stabilization at an elevated plateau. The response was not affected by reduction of extracellular Ca2+ concentration. The elevation in [Ca2+]i was canceled by pretreatment with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-AM, thapsigargin, and a phospholipase C inhibitor, U-73122. Under those experimental conditions, forskolin raised the beat frequency to a moderately elevated plateau, whereas the initial strong rise in frequency was completely abolished. All effects were canceled by H-89, a selective protein kinase A (PKA) inhibitor. The results suggest a dual role for PKA in ciliary regulation. PKA releases Ca2+ from intracellular stores, strongly activating ciliary beating, and, concurrently, produces moderate prolonged enhancement of the beat frequency by a Ca2+ independent mechanism. PMID- 9730964 TI - Rho GTPase signaling regulates tight junction assembly and protects tight junctions during ATP depletion. AB - Tight junctions control paracellular permeability and cell polarity. Rho GTPase regulates tight junction assembly, and ATP depletion of Madin-Darby canine kidney (MDCK) cells (an in vitro model of renal ischemia) disrupts tight junctions. The relationship between Rho GTPase signaling and ATP depletion was examined. Rho inhibition resulted in decreased localization of zonula occludens-1 (ZO-1) and occludin at cell junctions; conversely, constitutive Rho signaling caused an accumulation of ZO-1 and occludin at cell junctions. Inhibiting Rho before ATP depletion resulted in more extensive loss of junctional components between transfected cells than control junctions, whereas cells expressing activated Rho better maintained junctions during ATP depletion than control cells. ATP depletion and Rho signaling altered phosphorylation signaling mechanisms. ZO-1 and occludin exhibited rapid decreases in phosphoamino acid content following ATP depletion, which was restored on recovery. Expression of Rho mutant proteins in MDCK cells also altered levels of occludin serine/threonine phosphorylation, indicating that occludin is a target for Rho signaling. We conclude that Rho GTPase signaling induces posttranslational effects on tight junction components. Our data also demonstrate that activating Rho signaling protects tight junctions from damage during ATP depletion. PMID- 9730965 TI - Effect of intracellular pH on acetylcholine-induced Ca2+ waves in mouse pancreatic acinar cells. AB - We have used fluo 3-loaded mouse pancreatic acinar cells to investigate the relationship between Ca2+ mobilization and intracellular pH (pHi). The Ca2+ mobilizing agonist ACh (500 nM) induced a Ca2+ release in the luminal cell pole followed by spreading of the Ca2+ signal toward the basolateral side with a mean speed of 16.1 +/- 0.3 micron/s. In the presence of an acidic pHi, achieved by blockade of the Na+/H+ exchanger or by incubation of the cells in a Na+-free buffer, a slower spreading of ACh-evoked Ca2+ waves was observed (7.2 +/- 0.6 micron/s and 7.5 +/- 0.3 micron/s, respectively). The effects of cytosolic acidification on the propagation rate of ACh-evoked Ca2+ waves were largely reversible and were not dependent on the presence of extracellular Ca2+. A reduction in the spreading speed of Ca2+ waves could also be observed by inhibition of the vacuolar H+-ATPase with bafilomycin A1 (11.1 +/- 0.6 micron/s), which did not lead to cytosolic acidification. In contrast, inhibition of the endoplasmic reticulum Ca2+-ATPase by 2,5-di-tert-butylhydroquinone led to faster spreading of the ACh-evoked Ca2+ signals (25.6 +/- 1.8 micron/s), which was also reduced by cytosolic acidification or treatment of the cells with bafilomycin A1. Cytosolic alkalinization had no effect on the spreading speed of the Ca2+ signals. The data suggest that the propagation rate of ACh-induced Ca2+ waves is decreased by inhibition of Ca2+ release from intracellular stores due to cytosolic acidification or to Ca2+ pool alkalinization and/or to a decrease in the proton gradient directed from the inositol 1,4, 5-trisphosphate-sensitive Ca2+ pool to the cytosol. PMID- 9730966 TI - Mechanical loading regulates expression of talin and its mRNA, which are concentrated at myotendinous junctions. AB - The hypothesis that mechanical loading regulates talin expression in developing and adult muscle was tested using in vitro and in vivo models. Talin was selected for study because it is a key structural link between the cytoskeleton and cell membrane. In the in vitro model, C2C12 myotubes were subjected to cyclic strains for 48 h. In the in vivo model, rat hindlimb muscles were unloaded for 10 days, then reloaded for 2 days. Cyclic loading of myotubes resulted in significant increases in the quantity of talin (68%) and its 190-kDa proteolytic fragment (70%), as well as talin mRNA (180%), relative to unloaded myotube cultures. Similarly, talin concentration and its mRNA increased by 68 and 136%, respectively, in soleus muscles reloaded for 2 days relative to ambulatory controls. Immunohistochemistry and in situ RT-PCR showed that talin and its mRNA are concentrated and colocalized at myotendinous junctions. Thus these findings indicate that increased mechanical loading promotes talin synthesis, which occurs principally at myotendinous junctions, according to talin mRNA distribution. PMID- 9730967 TI - Mechanism of depression in cardiac sarcolemmal Na+-K+-ATPase by hypochlorous acid. AB - Oxidative stress during pathological conditions such as ischemia-reperfusion is known to promote the formation of hypochlorous acid (HOCl) in the heart and to result in depression of cardiac sarcolemmal (SL) Na+-K+-ATPase activity. In this study, we examined the direct effects of HOCl on SL Na+-K+-ATPase from porcine heart. HOCl decreased SL Na+-K+-ATPase activity in a concentration- and time dependent manner. Characterization of Na+-K+-ATPase activity in the presence of different concentrations of MgATP revealed a decrease in the maximal velocity (Vmax) value, without a change in affinity for MgATP on treatment of SL membranes with 0.1 mM HOCl. The Vmax value of Na+-K+-ATPase, when determined in the presence of different concentrations of Na+, was also decreased, but affinity for Na+ was increased when treated with HOCl. Formation of acylphosphate by SL Na+-K+ ATPase was not affected by HOCl. Scatchard plot analysis of [3H]ouabain binding data indicated no significant change in the affinity or maximum binding capacity value for ouabain binding following treatment of SL membranes with HOCl. Western blot analysis of Na+-K+-ATPase subunits in HOCl-treated SL membranes showed a decrease (34 +/- 9% of control) in the beta1-subunit without any change in the alpha1- or alpha2-subunits. These data suggest that the HOCl-induced decrease in SL Na+-K+-ATPase activity may be due to a depression in the beta1-subunit of the enzyme. PMID- 9730968 TI - Effect of Bcl-2 on oxidant-induced cell death and intracellular Ca2+ mobilization. AB - The mechanism by which Bcl-2 inhibits cell death is unknown. It has been suggested that Bcl-2 functions as an antioxidant. Because Bcl-2 is localized mainly to the membranes of the endoplasmic reticulum (ER) and the mitochondria, which represent the main intracellular storage sites for Ca2+, we hypothesized that Bcl-2 might protect cells against oxidant injury by altering intracellular Ca2+ homeostasis. To test this hypothesis, we examined the effect of oxidant treatment on viability in normal rat kidney (NRK) cells and in NRK cells stably transfected with Bcl-2 in the presence or absence of intracellular Ca2+, and we compared the effect of Bcl-2 expression on oxidant-induced intracellular Ca2+ mobilization and on ER and mitochondrial Ca2+ pools. NRK cells transfected with Bcl-2 (NRK-Bcl-2) were significantly more resistant to H2O2-induced cytotoxicity than control cells. EGTA-AM, an intracellular Ca2+ chelator, as well as the absence of Ca2+ in the medium, reduced H2O2-induced cytotoxicity in both cell lines. Compared with controls, cells overexpressing Bcl-2 showed a delayed rise in intracellular Ca2+ concentration ([Ca2+]i) after H2O2 treatment. After treatment with the Ca2+ ionophore ionomycin, Bcl-2-transfected cells showed a much quicker decrease after the maximal rise than control cells, suggesting stronger intracellular Ca2+ buffering, whereas treatment with thapsigargin, an inhibitor of the ER Ca2+-ATPases, transiently increased [Ca2+]i in control and in Bcl-2-transfected cells. Estimates of mitochondrial Ca2+ stores using an uncoupler of oxidative phosphorylation show that NRK-Bcl-2 cells have a higher capacity for mitochondrial Ca2+ storage than control cells. In conclusion, Bcl-2 may prevent oxidant-induced cell death, in part, by increasing the capacity of mitochondria to store Ca2+. PMID- 9730969 TI - Tissue distribution of immunoreactive mouse extracellular superoxide dismutase. AB - Protein content and mRNA expression of extracellular superoxide dismutase (EC SOD) were investigated in 16 mouse tissues. We developed a double-antibody sandwich ELISA using the affinity-purified IgG against native mouse EC-SOD. EC SOD could be detected in all of the tissues examined (lung, kidney, testis, brown fat, liver, adrenal gland, pancreas, colon, white fat, thymus, stomach, spleen, heart, skeletal muscle, ileum, and brain, in decreasing order of content measured as microg/g wet tissue). Lung showed a markedly higher value of EC-SOD than other tissues. Interestingly, white fat had a high content of EC-SOD in terms of micrograms per milligram protein, which corresponded to that of lung. Kidney showed the strongest expression of EC-SOD mRNA. Relatively strong expression of the mRNA was observed in lung, white fat, adrenal gland, brown fat, and testis. Heart and brain showed only weak signals, and no such expression could be detected in either digestive organs or skeletal muscle. Immunohistochemically, EC SOD was localized mainly to connective tissues and vascular walls in the tissues examined. Deep staining in the cytosol was observed in the cortical tubular cells of kidney. These results suggest that EC-SOD is distributed systemically in mice and that the physiological importance of this enzyme may be a compensatory adaptation to oxidative stress, particularly in lung and kidney. PMID- 9730970 TI - Characterization of the cloned human intermediate-conductance Ca2+-activated K+ channel. AB - The human intermediate-conductance, Ca2+-activated K+ channel (hIK) was identified by searching the expressed sequence tag database. hIK was found to be identical to two recently cloned K+ channels, hSK4 and hIK1. RNA dot blot analysis showed a widespread tissue expression, with the highest levels in salivary gland, placenta, trachea, and lung. With use of fluorescent in situ hybridization and radiation hybrid mapping, hIK mapped to chromosome 19q13.2 in the same region as the disease Diamond-Blackfan anemia. Stable expression of hIK in HEK-293 cells revealed single Ca2+-activated K+ channels exhibiting weak inward rectification (30 and 11 pS at -100 and +100 mV, respectively). Whole cell recordings showed a noninactivating, inwardly rectifying K+ conductance. Ionic selectivity estimated from bi-ionic reversal potentials gave the permeability (PK/PX) sequence K+ = Rb+ (1.0) > Cs+ (10.4) >> Na+, Li+, N-methyl-D-glucamine (>51). NH+4 blocked the channel completely. hIK was blocked by the classical inhibitors of the Gardos channel charybdotoxin (IC50 28 nM) and clotrimazole (IC50 153 nM) as well as by nitrendipine (IC50 27 nM), Stichodactyla toxin (IC50 291 nM), margatoxin (IC50 459 nM), miconazole (IC50 785 nM), econazole (IC50 2.4 microM), and cetiedil (IC50 79 microM). Finally, 1-ethyl-2-benzimidazolinone, an opener of the T84 cell IK channel, activated hIK with an EC50 of 74 microM. PMID- 9730971 TI - The opt1 gene of Drosophila melanogaster encodes a proton-dependent dipeptide transporter. AB - We have cloned and characterized the opt1 gene of Drosophila melanogaster. This gene encodes a protein with significant similarity to the PTR family of oligopeptide transporters. The OPT1 protein is localized to the apical epithelial membrane domains of the midgut, rectum, and female reproductive tract. The opt1 message is maternally loaded into developing oocytes, and OPT1 is found in the alpha-yolk spheres of the developing embryo. It is also found throughout the neuropil of the central nervous system, with elevated expression within the alpha and beta-lobes of the mushroom bodies. Transport activity was examined in HeLa cells transiently expressing OPT1. This protein is a high-affinity transporter of alanylalanine; the approximate Km constant is 48.8 microM for this substrate. OPT1 dipeptide transport activity is proton dependent. The ability of selected beta-lactams to inhibit alanylalanine transport suggests that OPT1 has a broad specificity in amino acid side chains and has a substrate requirement for an alpha-amino group. Together these data suggest an important role for OPT1 in regulating amino acid availability. PMID- 9730972 TI - Control of AMP deaminase 1 binding to myosin heavy chain. AB - AMP deaminase (AMPD) plays a central role in preserving the adenylate energy charge in myocytes following exercise and in producing intermediates for the citric acid cycle in muscle. Prior studies have demonstrated that AMPD1 binds to myosin heavy chain (MHC) in vitro; binding to the myofibril varies with the state of muscle contraction in vivo, and binding of AMPD1 to MHC is required for activation of this enzyme in myocytes. The present study has identified three domains in AMPD1 that influence binding of this enzyme to MHC using a cotransfection model that permits assessment of mutations introduced into the AMPD1 peptide. One domain that encompasses residues 178-333 of this 727-amino acid peptide is essential for binding of AMPD1 to MHC. This region of AMPD1 shares sequence similarity with several regions of titin, another MHC binding protein. Two additional domains regulate binding of this peptide to MHC in response to intracellular and extracellular signals. A nucleotide binding site, which is located at residues 660-674, controls binding of AMPD1 to MHC in response to changes in intracellular ATP concentration. Deletion analyses demonstrate that the amino-terminal 65 residues of AMPD1 play a critical role in modulating the sensitivity to ATP-induced inhibition of MHC binding. Alternative splicing of the AMPD1 gene product, which alters the sequence of residues 8-12, produces two AMPD1 isoforms that exhibit different MHC binding properties in the presence of ATP. These findings are discussed in the context of the various roles proposed for AMPD in energy production in the myocyte. PMID- 9730973 TI - Dichotomous development of the organic anion transport protein in liver and choroid plexus. AB - Both adult liver and choroid plexus express the organic anion transport protein (oatp1) and transport [35S]bromosulfophthalein (BSP). Studies of the developing rat liver reveal that oatp1 mRNA and protein do not begin to be expressed until 15 days postnatal and are at adult levels by 30 days. Uptake of [35S]BSP follows the same time course. In contrast, neonatal rat choroid plexus expresses oatp1 mRNA and protein. When quantified on a weight basis, the uptake of [35S]BSP in choroid plexus is lower in the adult than at earlier stages of development. Although fluorescence confocal microscopy of adult rat choroid plexus shows that oatp is localized to the apical surface, facing the cerebrospinal fluid, this method reveals an intracellular localization of oatp1 in the neonate. Approximately 12 wk are required for the appearance of the adult pattern of distribution. Changes in the localization and activity of oatp1 during development could play an important role in the pathobiology of maturation of the liver and the central nervous system. PMID- 9730975 TI - Correction for inner filter effects in turbid samples: fluorescence assays of mitochondrial NADH. AB - Fluorescent determinations of NADH in porcine heart mitochondria were subject to significant errors caused by alterations in inner filter effects during numerous metabolic perturbations. These inner filter effects were primarily associated with changes in mitochondrial volume and accompanying light scattering. The observed effects were detected in a standard commercial fluorometer with emission orthogonal to the excitation light path and, to a lesser extent, in a light path geometry detecting only the surface fluorescence. A method was developed to detect and correct for inner filter effects on mitochondrial NADH fluorescence measurements that were independent of the optical path geometry using an internal fluorescent standard and linear least-squares spectral analysis. A simple linear correction with the inner fluorescence reference was found to adequately correct for inner filter effects. This approach may be useful for other fluorescence probes in isolated mitochondria or other light-scattering media. PMID- 9730974 TI - Estrogen increases the permeability of the cultured human cervical epithelium by modulating cell deformability. AB - Estrogens increase secretion of cervical mucus in females. The objective of this research was to study the mechanisms of estrogen action. The experimental models were human CaSki (endocervical) and hECE (ectocervical) epithelial cells cultured on filters. Incubation in steroid-free medium increased transepithelial electrical resistance (RTE) and decreased epithelial permeability to the cell impermeant acid pyranine. Estrogen treatment reversed the effects, indicating estrogen decreases epithelial paracellular resistance. The estrogen effect was time and dose related (EC50 approximately 1 nM) and specific (estradiol = diethylstilbestrol > estrone, estriol; no effect by progesterone, testosterone, or cortisol) and was blocked by progesterone, tamoxifen, and ICI-182780 (an estrogen receptor antagonist). Estrogen treatment did not modulate dilution potential or changes in RTE in response to diC8 or to low extracellular Ca2+ (modulators of tight junctional resistance). In contrast, estrogen augmented decreases in RTE in response to hydrostatic and hypertonic gradients [modulators of resistance of lateral intercellular space (RLIS)], suggesting estrogen decreases RLIS. Estrogen decreased cervical cell size, shortened response time relative to changes in cell size after hypertonic challenge, and augmented the decrease in cell size in response to hypertonic and hydrostatic gradients. Lowering luminal NaCl had no significant effect on RTE, and the Cl- channel blocker diphenylamine-2-carboxylate attenuated the hypertonicity-induced decrease in cell size to the same degree in control and estrogen-treated cells, suggesting estrogen effects on permeability and cell size are not mediated by modulating Na+ or Cl- transport. In contrast, estrogen increased cellular G-actin levels, suggesting estrogens shift actin steady-state toward G-actin and the cervical cell cytoskeleton toward a more flexible structure. We suggest that the mechanism by which estrogens decrease RLIS and increase permeability is by fragmenting the cytoskeleton and facilitating deformability and decreases in cervical cell size. PMID- 9730976 TI - Protein transport from the endoplasmic reticulum to the Golgi apparatus. AB - As the first step of protein transport along the biosynthetic (secretory/exocytotic) pathway, transport from the endoplasmic reticulum (ER) to the Golgi apparatus has received much attention over the past several decades. The general structural organization underlying this transport process is becoming more defined. The major protein components participating in the budding, pre docking, and docking/fusion events have been identified and their mechanistic aspects investigated. Conceptually, it is now clear that protein export from the ER is a selective process. Although much remains to be defined or refined, the general picture of this transport step has now emerged. PMID- 9730977 TI - Assembly of nuclear pore complexes and annulate lamellae promotes normal pronuclear development in fertilized mammalian oocytes. AB - In addition to functional nuclear pore complexes engaged in nucleo-cytoplasmic transport, the cytoplasmic stacks of pore complexes, called annulate lamellae, exist in numerous cell types. Although both annulate lamellae and nuclear pore complexes are present in fertilized mammalian oocytes, their relative roles in the process of fertilization and preimplantation development are not known. Using epifluorescence and electron microscopy, we explored their fate during bovine fertilization. The assembly of annulate lamellae in bovine oocytes was triggered by sperm-oocyte binding and continued concomitantly with the incorporation of the nuclear pores in the nuclear envelopes of the developing male and female pronuclei. This process was also induced by the parthenogenetic activation of metaphase-II-arrested oocytes. Depletion of Ca2+, previously implicated in oocyte activation and in the insertion of pore complexes into the nuclear envelope, prevented the formation of nuclear pore complexes, but not the assembly of annulate lamellae in oocyte cytoplasm. Injection of the nuclear pore antagonist, wheat germ agglutinin, into the cytoplasm of mature oocytes that were subsequently fertilized caused the arrest of pronuclear development, indicating the requirement of nuclear pore complexes for normal pronuclear development. Treatment of the fertilized oocytes with the microtubule inhibitor, nocodazole, prevented gathering of annulate lamellae around the developing pronuclei, insertion of nuclear pores into their nuclear envelopes, and further pronuclear development. The formation of the male pronuclei was reconstituted in Xenopus egg extracts and reflected the behavior of nuclear pores during natural fertilization. These data suggest that nuclear pore complexes are required for normal pronuclear development from its beginning up until pronuclear apposition. Annulate lamellae may be involved in the turnover of nuclear pore complexes during fertilization, which is in turn facilitated by the reorganization of oocyte microtubules and influx of Ca2+ into oocyte cytoplasm. PMID- 9730978 TI - Localisation of Nramp1 in macrophages: modulation with activation and infection. AB - The murine natural resistance-associated macrophage protein, Nramp1, has multiple pleiotropic effects on macrophage activation and regulates survival of intracellular pathogens including Leishmania, Salmonella and Mycobacterium species. Nramp1 acts as an iron transporter, but precisely how this relates to macrophage activation and/or pathogen survival remains unclear. To gain insight into function, anti-Nramp1 monoclonal and polyclonal antibodies are used here to localise Nramp1 following activation and infection. Confocal microscope analysis in uninfected macrophages demonstrates that both the mutant (infection susceptible) and wild-type (infection-resistant) forms of the protein localise to the membranes of intracellular vesicular compartments. Gold labelling and electron microscopy defines these compartments more precisely as electron-lucent late endosomal and electron-dense lysosomal compartments, with Nramp1 colocalizing with Lamp1 and cathepsins D and L in both compartments, with macrosialin in late endosomes, and with BSA-5 nm gold in pre-loaded lysosomes. Nramp1 is upregulated with interferon-(gamma) and lipopolysaccaride treatment, coinciding with an increase in labelling in lysosomes relative to late endosomes and apparent dispersion of Nramp1-positive vesicles from a perinuclear location towards the periphery of the cytoplasm along the microtubular network. In both control and activated macrophages, expression of the protein is 3- to 4-fold higher in wild-type compared to mutant macrophages. In Leishmania major-infected macrophages, Nramp1 is observed in the membrane of the pathogen-containing phagosomes, which retain a perinuclear localization in resting macrophages. In Mycobacterium avium-infected resting and activated macrophages, Nramp1-positive vesicles migrated to converge, but not always fuse, with pathogen-containing phagosomes. The Nramp1 protein is thus located where it can have a direct influence on phagosome fusion and the microenvironment of the pathogen, as well as in the more general regulation of endosomal/lysosomal function in macrophages. PMID- 9730979 TI - Strategies of epithelial repair: modulation of stem cell and transit amplifying cell proliferation. AB - Using double labeling techniques, we studied the replication of corneal epithelial stem cells that reside exclusively in the limbal zone, and their progeny transit amplifying cells. We show that corneal epithelial stem cells can be induced to enter DNA synthesis by wounding and by TPA. We demonstrate the existence of a hierarchy of TA cells; those of peripheral cornea undergo at least two rounds of DNA synthesis before they become post-mitotic, whereas those of central cornea are capable of only one round of division. However, the cell cycle time of these TA cells can be shortened and the number of times these TA cells can replicate is increased in response to wounding. These results thus demonstrate three strategies of epithelial repair: (i) stem cell replication, (ii) the unleashing of additional rounds of cell proliferation that remain as an untapped reserve under normal circumstances, and (iii) enhancement of TA cell proliferation via a shortening of the cycling time. PMID- 9730980 TI - Identification of cofilin, coronin, Rac and capZ in actin tails using a Listeria affinity approach. AB - Actin assembly is involved in cell motility and intracellular movement of Listeria monocytogenes. Induction of Listeria actin tails is mediated by the surface protein ActA. The N-terminal domain of ActA is sufficient for this function. Cell components known to play a role in the actin-based motility of Listeria are VASP (vasodilatator-stimulated phosphoprotein), the multiprotein Arp2/3 complex and cofilin. VASP interacts with the central domain of ActA. Proteins interacting with the N-terminal domain of ActA have not been identified. To identify novel host cell components of ActA-induced actin tails, we used bovine brain extracts and an affinity approach with Listeria as matrix. Several known components of Listeria tails were isolated including VASP, Arp3 and cofilin. Cofilin was identified by peptide sequencing, and cofilin recruitment and Listeria tail length were found to be pH-dependent, in agreement with its recently reported role in enhancing actin filament turnover. In addition, three proteins not previously known to be associated with Listeria tails, coronin, Rac and capZ, were identified in our affinity approach. In infected cells, the localization of the identified proteins was studied by immunofluorescence. Our findings suggest that these latter proteins, which are known to play critical roles in cellular actin rearrangements, may also be involved in the dynamics of Listeria-induced actin assembly. PMID- 9730981 TI - Muscle-specific functions of ryanodine receptor channels in Caenorhabditis elegans. AB - Ryanodine receptor channels regulate contraction of striated muscle by gating the release of calcium ions from the sarcoplasmic reticulum. Ryanodine receptors are expressed in excitable and non-excitable cells of numerous species, including the nematode C. elegans. Unlike vertebrates, which have at least three ryanodine receptor genes, C. elegans has a single gene encoded by the unc-68 locus. We show that unc-68 is expressed in most muscle cells, and that the phenotypic defects exhibited by unc-68 null mutants result from the loss of unc-68 function in pharyngeal and body-wall muscle cells. The loss of unc-68 function in the isthmus and terminal bulb muscles of the pharynx causes a reduction in growth rate and brood size. unc-68 null mutants exhibit defective pharyngeal pumping (feeding) and have abnormal vacuoles in the terminal bulb of the pharynx. unc-68 is required in body-wall muscle cells for normal motility. We show that UNC-68 is localized in body-wall muscle cells to flattened vesicular sacs positioned between the apical plasma membrane and the myofilament lattice. In unc-68 mutants, the vesicles are enlarged and densely stained. The flattened vesicles in body-wall muscle cells thus represent the C. elegans sarcoplasmic reticulum. Morphological and behavioral phenotypes of unc-68 mutants suggest that intracellular calcium release is not essential for excitation-contraction coupling in C. elegans. PMID- 9730982 TI - The Drosophila POLO kinase localises to multiple compartments of the mitotic apparatus and is required for the phosphorylation of MPM2 reactive epitopes. AB - The MPM2 antibody is a valuable tool for studying the regulation of mitotic events since it specifically recognises a subset of mitosis-specific phosphoproteins. Some MPM2 epitopes have been shown to be phosphorylated by p34(cdc2). However, recent results suggest that the newly emerging family of polo like kinases (Plks) may also act as MPM2 kinases. In this study, we present evidence suggesting that the Drosophila POLO protein is required for the phosphorylation of MPM2 reactive epitopes. POLO displays a dynamic localisation pattern during mitosis, which parallels that of the MPM2 phosphoepitopes, since it is found in the centrosome and centromere from early prophase until late anaphase, the microtubule-overlapping region during anaphase, and the region on either side of the midbody during telophase. Centromere localisation is not dependent upon microtubules since it is retained in colchicine-arrested cells and is present in isolated chromosomes. Furthermore, the level of MPM2 immunoreactivity is directly correlated to the severity of the polo mutant alleles. In cells carrying a hypomorphic allele, the centrosomes of abnormal cells are small and fail to efficiently recruit MPM2 epitopes. In neuroblasts homozygous for a severe loss-of-function allele, the mitotic index is low and the MPM2 labelling is severely reduced or absent. Finally, rephosphorylation of MPM2 epitopes in detergent-extracted Schneider cells requires either POLO stably bound to the cytoskeletons or POLO present in soluble extracts. These results suggest that POLO is required for the phosphorylation of MPM2 epitopes in Drosophila, at the centrosomes, centromeres and the mitotic spindle, and thus might be involved in co-ordinating the mitotic changes of cellular architecture with the activity of the maturation promoting factor. PMID- 9730983 TI - The actin-myosin cytoskeleton mediates reversible agonist-induced membrane blebbing. AB - Suprastimulation of pancreatic acinar cells with specific agonists inhibits zymogen secretion and induces the formation of large basolateral blebs. Currently the molecular mechanisms that mediate this dramatic morphologic response are undefined. Further, it is unclear if blebbing represents a terminal or reversible event. Using computer-enhanced video microscopy of living acini we have found that these large blebs form rapidly (within 2-3 minutes) and exhibit ameboid undulations. They are induced by small increases in agonist concentration and require an energy-dependent phosphorylation event. Remarkably, the blebs are rapidly absorbed when agonist levels are reduced, indicating that blebbing is a reversible response to a physiological stimulus, not a terminal event. Morphological methods show that these dramatic changes in cell shape are accompanied by a marked reorganization of actin and myosin II at the basolateral domain. During 30 minutes of suprastimulation, both basolateral actin and myosin II gradually increase to form a ring centered at the necks of the blebs. Immunocytochemical and biochemical studies with a phospho-specific antibody to the myosin regulatory light chain reveal an activation of myosin II in suprastimulated acini that is completely absent in resting cells. Studies using cytoskeletal antagonistic drugs indicate that bleb formation and motility require actin remodeling concomitant with an activation of myosin II. This aberrant activation and reorganization of the actin-myosin cytoskeleton is likely to have detrimental effects on acinar cell function. Additionally, this mechanism of bleb formation may be conserved among other forms of physiological blebbing events. PMID- 9730984 TI - High levels of actin tyrosine phosphorylation: correlation with the dormant state of Dictyostelium spores. AB - Upon removal of nutrients, the amoebae of the cellular slime mold Dictyostelium discoideum differentiate into dormant spores which survive starvation stress. In this study, we demonstrate that half of the actin molecules in the spores are tyrosine-phosphorylated. The phosphorylated actin is distributed around immobile crenate mitochondria and vesicles, as well as in the cytoplasm of the spores. The actin isolated from spore lysates contains phosphorylated and unphosphorylated forms at the same molar ratio as that of the original whole spore lysate. Under actin polymerizing conditions they form actin filaments and then they are completely depolymerized under actin depolymerizing conditions, indicating that tyrosine phosphorylation of actin may not prohibit actin polymerization nor stimulate depolymerization. The phosphorylation levels increase at the end of the culmination stage when spores have matured morphologically and physiologically, and reach maximum levels after an additional 12 hours of development. The levels are stable for 20 days following spore maturation, and decline to undetectable levels within the next 10 days. Spores having high levels of phosphorylation show high viability, and vice versa. Following activation of spores with nutrient medium containing spore germination promoters, the phosphorylation levels quickly decrease with a half-life of about 5 minutes. After 20 minutes spores begin to swell. At this later time, most of the phosphorylated actin already has been dephosphorylated. Also, in heat-activated spores actin dephosphorylation occurs prior to spore swelling. However, addition of phosphatase inhibitors following heat-activation, prevented spore swelling and dephosphorylation of actin. Our data indicate that the high levels of actin tyrosine phosphorylation, specific to the spore stage, may be required for maintaining dormancy to withstand starvation stress. The rapid dephosphorylation of actin leads to a reactivated dynamic actin system which participates in spore swelling, vesicle movement, and mitochondrial shape changes during the spore germination process. PMID- 9730985 TI - Fibronectin matrix assembly enhances adhesion-dependent cell growth. AB - Cell growth control in non-transformed cells depends, in part, on adhesive interactions with the extracellular matrix. Following injury, excess or altered fibronectin deposition into the extracellular matrix may contribute to the pathogenesis of fibrosis and atherosclerosis by triggering changes in specific cell functions associated with wound repair, including cell proliferation and migration. To assess the role of fibronectin polymerization on cell growth, we isolated mouse embryonic cells that lack endogenous fibronectin (fibronectin-null cells) and established them in culture under serum-free conditions. These fibronectin-null cells do not produce any detectable fibronectin, but are capable of assembling a fibronectin matrix when cultured in the presence of exogenously added fibronectin. Our data indicate that adhesion-dependent growth in fibronectin-null cells is dramatically increased (>2-5x) by culturing cells in the presence of fibronectin. This fibronectin-induced cell growth was blocked by inhibiting fibronectin matrix assembly. Arg-Gly-Asp peptides or fragments of fibronectin that contain the Arg-Gly-Asp cell binding site promoted clustering of the (&agr ;)5beta1 integrin in focal adhesions, but did not enhance cell growth. These data indicate that the polymerization of fibronectin into the extracellular matrix positively regulates cell growth, and that occupancy and clustering of fibronectin-binding integrins alone are not sufficient to trigger increased cell growth. PMID- 9730986 TI - Embryoglycan ectodomains regulate biological activity of FGF-2 to embryonic stem cells. AB - Basic fibroblast growth factor (FGF-2) functions as a natural inducer of mesoderm, regulator of cell differentiation and autocrine modulator of cell growth and transformation. The FGF-2 signals are transduced through receptors with intrinsic protein tyrosine kinase activity. However, receptor binding and activation is governed by extracellular matrix, cell surface or soluble proteoglycans. This paper focuses on the role of proteoglycans synthesized by embryonic cells, embryoglycans, in FGF-2 signaling via FGF receptor-1 (FGFR-1). We found that embryoglycan ectodomain Lewis X, analog of developmentally regulated embryonic cell surface epitope TEC 1, promotes oligomerization of FGF-2 in the cell free chemical crosslinking. In vitro assays show that a large molar excess of extracellular Lewis X does not inhibit binding of FGF-2 to embryonic stem (ES) cells, but prevents the mitogenic effect of FGF-2. Western blot analysis of ES cells revealed the presence of abundant 52 kDa and trace amounts of 67 and 125 kDa isoforms of FGFR-1. However, none of these isoforms undergo any detectable changes in tyrosine phosphorylation under the conditions that modulate the mitogenic effect of FGF-2. Rather, a primary substrate of all receptor tyrosine kinases, phospholipase C gamma (PLC gamma), is activated by both FGF-2 and Lewis X. The combination, FGF-2 plus Lewis X, leads to weak inhibition, when compared with the effects of FGF-2 and Lewis X, respectively. In accordance, the level of phosphorylation of non-receptor tyrosine kinase c-Src is reduced in a reversed pattern to PLC(gamma). Furthermore, in this particular cell type we show the presence of activated forms of extracellular signal-related kinase (ERK) in all nontreated and treated cells. These findings demonstrate that embryoglycan ectodomains may act as negative regulators of FGF-2-induced ES cell proliferation, most likely through the FGFR-1-independent signaling pathway. PMID- 9730987 TI - Anosmin-1 underlying the X chromosome-linked Kallmann syndrome is an adhesion molecule that can modulate neurite growth in a cell-type specific manner. AB - Anosmin-1 is an extracellular matrix glycoprotein which underlies the X chromosome-linked form of Kallmann syndrome. This disease is characterized by hypogonadism due to GnRH deficiency, and a defective sense of smell related to the underdevelopment of the olfactory bulbs. This study reports that anosmin-1 is an adhesion molecule for a variety of neuronal and non-neuronal cell types in vitro. We show that cell adhesion to anosmin-1 is dependent on the presence of heparan sulfate and chondroitin sulfate glycosaminoglycans at the cell surface. A major cell adhesion site of anosmin-1 was identified in a 32 amino acid (32R1) sequence located within the first fibronectin-like type III repeat of the protein. The role of anosmin-1 as a substrate for neurite growth was tested on either coated culture dishes or monolayers of anosmin-1-producing CHO cells. In both experimental systems, anosmin-1 was shown to be a permissive substrate for the neurite growth of different types of neurons. Mouse P5 cerebellar neurons cultured on anosmin-1 coated wells developed long neurites; the 32R1 peptide was found to underly part of this neurite growth activity. When the cerebellar neurons were cultured on anosmin-1-producing CHO cells, neurite growth was reduced as compared to wild-type CHO cells; in contrast, no difference was observed for E18 hippocampal and P1 dorsal root ganglion neurons in the same experimental system. These results indicate that anosmin-1 can modulate neurite growth in a cell-type specific manner. Finally, anosmin-1 induced neurite fasciculation of P5 cerebellar neuron aggregates cultured on anosmin-1-producing CHO cells. The pathogenesis of the olfactory defect in the X-linked Kallmann syndrome is discussed in the light of the present results and the recent data reporting the immunohistochemical localisation of anosmin-1 during early embryonic development. PMID- 9730988 TI - Common and variant properties of intermediate filament proteins from lower chordates and vertebrates; two proteins from the tunicate Styela and the identification of a type III homologue. AB - The chordates combine the vertebrates and the invertebrate phyla of the cephalo- and urochordates (tunicates). Two cytoplasmic intermediate filament (IF) proteins of the urochordate Styela plicata are characterized by cDNA cloning, gene organization, tissue specific expression patterns in the adult animal and the self assembly properties of the recombinant proteins. In line with metazoan phylogeny St-A and St-B have the short length version of the coil 1b domain found in all vertebrate and cephalochordate IF proteins while protostomic IF proteins have the longer length version with an extra 42 residues. St-A is the first IF protein from a lower chordate which can be unambiguously related to a particular vertebrate IF subfamily. St-A shares 46% sequence identity with desmin, displays the N-terminal motif necessary for filament assembly of type III proteins and forms normal homopolymeric 10 nm filaments in vitro. St-A but not St-B is present in smooth muscle cells of the body wall musculature. St-A and St-B are found as separate networks in some interior epithelia. St-B shares 30 to 35% identity with keratin 8, St-A and desmin and does not form IF under in vitro assembly conditions. Its relation to a particular vertebrate IF type or to the eight currently known IF proteins from the cephalochordate Branchiostoma remains unresolved. The striking relation between St-A and desmin predicts that the common progenitor of the urochordate (tunicate) and the cephalochordate/vertebrate lineages already possessed a type III homologue. Unlike in vertebrates intron patterns cannot be used to classify the tunicate IF genes. Although St-A is a type III homologue its gene shows an intron position which in vertebrates is restricted to keratin type II genes. PMID- 9730989 TI - Transforming growth factor-beta dynamically regulates vascular smooth muscle differentiation in vivo. AB - Variations in the levels of smooth muscle-specific isoforms of contractile proteins have been reported to occur in many different vascular diseases. However, although much work has been done in vitro to investigate the regulation of smooth muscle cell differentiation, the molecular mechanisms which regulate the differentiation of vascular smooth muscle tissue in vivo are unknown. Using quantitative immunofluorescence, we show that in rat arteries levels of smooth muscle differentiation markers correlate with the levels of the cytokine TGF beta. In young mice with one allele of the TGF-beta1 gene deleted, the levels of both TGF-beta1 and smooth muscle differentiation markers are reduced compared to wild-type controls. This regulation of smooth muscle differentiation by TGF-beta during post-natal development also occurs dynamically in the adult animal. Following various pharmacological or surgical interventions, including treatment of mice with tamoxifen and balloon injury of rat carotid arteries, there is a strong correlation between the changes in the levels of TGF-beta and changes in the levels of smooth muscle differentiation markers (r=0. 9, P<0.0001 for n=26 experiments). We conclude that TGF-beta dynamically regulates smooth muscle differentiation in rodent arteries in vivo. PMID- 9730990 TI - Regulation of mammalian replication origin usage in Xenopus egg extract. AB - Xenopus embryos initiate replication at random closely spaced sites until a certain concentration of nuclei is achieved within the embryo, after which fewer, more specific chromosomal sites are utilized as origins. We have examined the relationship between nucleo-cytosolic ratio and origin specification when Chinese hamster ovary (CHO) cell nuclei are introduced into Xenopus egg extracts. At concentrations of intact late-G1-phase nuclei that approximate early Xenopus embryos, the entire genome was duplicated nearly 4 times faster than in culture, accompanied by a de-localization of initiation sites at the dihydrofolate reductase (DHFR) locus. As the concentration of nuclei was increased, the number of initiation sites per nucleus decreased and initiation at the DHFR locus became localized to the physiologically utilized DHFR origin. Origin specification was optimal at nuclear concentrations that approximate the Xenopus mid-blastula transition (MBT). Higher concentrations resulted in an overall inhibition of DNA synthesis. By contrast, with intact early G1-phase nuclei, replication initiated at apparently random sites at all concentrations, despite an identical relationship between nucleo-cytosolic ratio and replicon size. Furthermore, permeabilization of late-G1-phase nuclei, using newly defined conditions that preserve the overall rate of replication, eliminated site-specificity, even at nuclear concentrations optimal for DHFR origin recognition. These data show that both nucleo-cytosolic ratio and nuclear structure play important but independent roles in the regulation of replication origin usage. Nucleo-cytosolic ratio clearly influences the number of replication origins selected. However, titration of cytosolic factors is not sufficient to focus initiation to specific sites. An independent mechanism, effecting changes within G1-phase nuclei, dictates which of many potential initiation sites will function as an origin. PMID- 9730991 TI - Maternal smoking and infant lung function. Further evidence for an in utero effect. PMID- 9730992 TI - Pharmacologic treatment of sleep-disordered breathing. AB - Available literature on the use of pharmacologic agents for the treatment of sleep-disordered breathing was reviewed by evidenced-based methodology. Evidence tables were created and studies were graded according to study design and the number of subjects included. Scores for each group of studies evaluating each pharmacologic agent were established so that the quality of research for different drugs could be compared. The use of various ventilatory stimulants, psychotropic drugs, and antihypertensive agents were reviewed. The most objective data are available on theophylline and opioid antagonist/nicotine groups. Although more controlled studies would be helpful, relatively clear-cut indications for the use of ventilatory stimulants exist for hypercapnic obesity hypoventilation patients (medroxyprogesterone), myxedema (thyroid replacement), central apnea (acetazolamide), and periodic breathing in congestive heart failure (theophylline). Few randomized, well-controlled trials have been published that evaluate pharmacologic agents in the treatment of classic OSA. To date, no one agent stands out as being useful for OSA. Future research will need to characterize subjects so that various subsets of patients can be tried on one or on a combination of various pharmacologic agents. PMID- 9730993 TI - Respiratory function among preterm infants whose mothers smoked during pregnancy. AB - We examined whether the adverse effects of prenatal exposure to tobacco on lung development are limited to the last weeks of gestation by comparing respiratory function in preterm infants whose mothers had and had not smoked during pregnancy. Maximal forced expiratory flow (Vmax FRC) and time to peak tidal expiratory flow as a proportion of total expiratory time (TPTEF:TE) were measured prior to discharge from hospital in 108 preterm infants (mean [SD] gestational age, 33.5 [1.8] wk), 40 of whose mothers had smoked during pregnancy. Infant urinary cotinine was less than 4 ng/ml in those born to nonsmokers, but it was as high as 458 ng/ml in exposed infants (p < 0.0001). TPTEF:TE was significantly lower in infants exposed to tobacco in utero (mean [SD], 0.369 [0.109]) when compared with those who were not (0.426 [0.135]) (p <= 0.02). Vmax FRC was also reduced in exposed infants (mean [SD], 85.2 [41.7] ml/s versus 103.8 [49.7] ml/s) (p = 0.07). After allowing for sex, ethnic group, body size, postnatal age, and socioeconomic status, TPTEF:TE remained significantly diminished in infants exposed prenatally to tobacco (p < 0.05). Thus, impaired respiratory function is evident in infants born on average 7 wk prior to the expected delivery date, suggesting that the adverse effects of prenatal exposure to tobacco are not limited to the last weeks of pregnancy. PMID- 9730994 TI - Pulmonary surfactant activity is impaired in lung transplant recipients. AB - Impaired graft function in the postoperative course after lung transplantation (LTx) may in part be due to alterations in pulmonary surfactant. Animal data provide increasing evidence for surfactant abnormalities in the early course after graft reperfusion. However, little is known about the integrity of the surfactant system in human lung transplant recipients. We therefore investigated surfactant properties in bronchoalveolar lavage fluid (BALF) of patients with lung transplants (n = 60) in comparison to that of healthy subjects (n = 10). The phospholipid concentrations of BALF and of surfactant subfractions were measured, and total protein was analyzed. Surface activity was measured with a pulsating bubble surfactometer (PBS). Minimum surface tension was 15.8 +/- 1.1 mN/m in lung transplant recipients, whereas healthy subjects had minimum surface tensions of 3.4 +/- 1.9 mN/m (p = 0.0004). As a marker for potential surfactant inhibition, protein-to-phospholipid (PL) ratios showed no significant differences in the two major study groups. The ratio of small surfactant aggregates to large surfactant aggregates was increased in patients with lung transplants (p = 0.043). Episodes of infection or rejection did not change surface activities, nor did they induce altered ratios of protein to PL or of small to large surfactant aggregates. Surfactant activity did not correlate with pulmonary-function data. Moreover, surface tension showed no correlation with the time after transplantation. Our results suggest a persistent impairment of biophysical surfactant properties after LTx, possibly due to type-II-cell malfunction. PMID- 9730995 TI - Assessment of inspiratory flow limitation invasively and noninvasively during sleep. AB - To define the standard of airway flow limitation, pharyngeal pressure and flow rate were measured during wakefulness and sleep in seven habitual snorers with widely varying degrees of sleep-induced increases in upper airway resistance. Inspiratory pressure:flow relationships were used to group breaths into four categories of flow limitation, including linear (Level 1), mildly alinear (Level 2), constant flow rate with no pressure dependence (Level 3), and decreasing flow rate throughout significant portions of inspiration, i.e., negative pressure dependence (Level 4). These pressure:flow rate gold standards of flow limitation were used to evaluate a flow limitation index derived from the time profile (or "shape") of three noninvasive estimates of flow rate: (1) pneumotach flow rate, (2) differentiated sum respiratory inductance plethysmography (RIP), and (3) nasal pressure. A nonflow limited template for each of these noninvasive measurements was taken from awake breaths and the difference in area determined between the template breath and each of the noninvasive signals measured during nonrapid eye movement (NREM) sleep. The noninvasive flow limitation indices were found to be effective in differentiating severe types of inspiratory flow limitation, i.e., Level 1 versus Level 3 or Level 4 (sensitivity/specificity > 80%). On the other hand, these indirect indices were not able to consistently detect mild levels of flow limitation (Level 1 versus Level 2; sensitivity/specificity = 62 to 72%); nor were these noninvasive estimates of flow rate "shape" sensitive to breaths with a high but fixed resistance throughout inspiration. The area index derived from measurements of pressure at the nares (Pn) was the most sensitive, nonperturbing, noninvasive measure of flow rate and flow limitation, and we recommend its use for recognizing most of the common types of moderate to severe levels of airway flow limitation in sleeping subjects. PMID- 9730996 TI - Inhaled fluticasone reduces sputum inflammatory indices in severe bronchiectasis. AB - Although corticosteroid therapy might be clinically beneficial for bronchiectasis, very little is known of its effects on the inflammatory and infective markers in bronchiectasis. We have therefore performed a double-blind, placebo-controlled study to evaluate the effects of a 4-wk administration of inhaled fluticasone in bronchiectasis. Twenty-four patients (12 female; mean age 51 yr) were randomized into receiving either inhaled fluticasone (500 microgram twice daily) via the Accuhaler device (n = 12) or placebo. At each visit, spirometry, 24-h sputum volume, sputum leukocyte density, bacterial densities, and concentrations of interleukin (IL)-1beta, IL-8, tumor necrosis factor-alpha (TNF-alpha), and leukotriene B4 (LTB4) were determined. There was a significant (p < 0.05) decrease in sputum leukocyte density and IL-1beta, IL-8, and LTB4 after fluticasone treatment. The fluticasone group had one and the placebo group three episodes of exacerbation. There were no significant changes in spirometry (p > 0.05) or any reported adverse reactions in either group. The results of this study show that high-dose fluticasone is effective in reducing the sputum inflammatory indices in bronchiectasis. Large-scale and long-term studies are indicated to evaluate the effects of inhaled steroid therapy on the inflammatory components in bronchiectasis. PMID- 9730997 TI - Effects of changes in fresh fruit consumption on ventilatory function in healthy British adults. AB - Cross-sectional studies have shown frequent fresh fruit consumption to be associated with higher lung function in both children and adults. This relationship is investigated longitudinally in a national sample of 2,171 British adults age 18 to 73 initially examined in 1984, who were reexamined 7 yr later, and had no reported history of chronic respiratory disease throughout. Outcome was assessed by change in forced expiratory volume in one second (FEV1) between the two examinations, standardized for age, height, and sex and related to fresh fruit consumption estimated by food frequency questionnaires at both examinations. After adjustment for region, social class, and smoking, changes in fresh fruit consumption levels were positively associated with changes in FEV1 (p = 0.002), highlighted by a more marked fall in FEV1 (107 ml; 95% confidence interval, 36 to 178 ml) in subjects who reduced their fresh fruit consumption the greatest compared with those with no change. In contrast, average levels of fruit intake were not associated with change in FEV1 (p = 0.695). The implication is that the cross-sectional effects of fresh fruit consumption on ventilatory function appear to be reversible and not progressive, such that consistently low levels of fresh fruit intake do not appear to increase lung function decline. PMID- 9730998 TI - Expiratory muscle function in amyotrophic lateral sclerosis. AB - Few data exist concerning expiratory muscle function in amyotrophic lateral sclerosis (ALS). We studied 26 patients with ALS (16 with respiratory symptoms and 10 without) and measured the maximal static expiratory mouth pressure (MEP), the gastric pressure during a maximal cough (Cough Pga), and the gastric pressure after magnetic stimulation of the lower thoracic nerve roots (Tw Pga). These measurements were related to the ability to generate transient supramaximal flow during a cough (cough spikes), to arterialized capillary blood gases, and to inspiratory muscle strength. Vocal cord motion was examined endoscopically in 11 of the 16 symptomatic patients. Expiratory muscle weakness was related to inability to generate cough spikes with a threshold effect such that spikes were absent for Cough Pga < 50 cm H2O (p = 0.009) or Tw Pga < 7 cm H2O (p = 0.006) and was usually associated with inspiratory muscle weakness. However, in multivariate analysis, PaCO2 was only significantly associated with the maximal sniff esophageal pressure (p = 0.02). Symptomatic patients had significantly lower inspiratory muscle strength, whereas, of the expiratory muscle tests, only Tw Pga was significantly lower (p = 0.0009) in symptomatic patients. Abnormal vocal cord motion was observed in two of the 11 patients examined. We conclude that abdominal muscle weakness in ALS, when substantial, results in an inability to generate transient supramaximal flow during a cough. However, the primary determinant of both ventilatory failure and respiratory symptoms seems to be inspiratory muscle weakness. PMID- 9730999 TI - Control of ventilation during lung volume changes and permissive hypercapnia in dogs. AB - We investigated the effect changes in end-expiratory lung volume (EEVL) had on the response to progressive hypercapnia (CO2-response curve) in eight open-chest, anesthetized dogs, in order to clarify the role that vagal lung mechanoreceptors have in altered respiratory drive during permissive hypercapnia. The dogs were ventilated using a positive-pressure ventilator driven by phrenic neural activity. Systemic arterial CO2 tension (PaCO2) was elevated by increasing the fraction of CO2 delivered to the ventilator. EEVL was altered from approximated functional residual capacity ("FRC") to 1.5 and 0.5 "FRC" by changing positive end-expiratory pressure. Although the tidal volume (VT)-PaCO2 and inspiratory time (TI)-PaCO2 relationships were not affected, decreasing EEVL from 1.5 "FRC" to "FRC" and then to 0.5 "FRC" caused a significant (p < 0.01) upward shift in the CO2-response curves for minute ventilation (V I) and frequency (f ), and a significant (p < 0.01) downward shift in the CO2- response curve for expiratory time (TE). We conclude that these shifts were explained by a decrease in the inhibitory activity of slowly adapting pulmonary stretch receptors (PSRs) as EEVL was lowered. In addition, increases in EEVL from 0.5 "FRC" to 1.5 "FRC" caused a significant (p < 0.05) increase in the apneic threshold, which we attribute to an inhibitory effect on central drive caused by increased PSR activity. PMID- 9731000 TI - Language of dyspnea in assessment of patients with acute asthma treated with nebulized albuterol. AB - To investigate whether the language of dyspnea provides relevant clinical information in addition to that provided by ratings of overall dyspnea intensity when assessing subjective response to therapy, we conducted a prospective study in a cohort of 25 patients with acute asthma presenting to the emergency department of a tertiary care hospital. Patients received nebulized albuterol treatments every 20 min with a maximum of three doses. At presentation and after each treatment, patients completed spirometry, rated overall dyspnea intensity on a modified Borg scale, and selected phrases that described qualities of breathlessness from a 15-item questionnaire. Paired Student's t tests revealed significant improvements in FEV1 (from 1.39 +/- 0.66 L to 1.80 +/- 0.76 L, p < 0. 001) and reductions in dyspnea intensity (from 5.12 +/- 2.08 to 2.82 +/- 1.59, p < 0.001) after the first albuterol treatment. Dyspnea intensity continued to decrease significantly in response to the second treatment, modified Borg rating 2.26 +/- 1.52, although there was no positive bronchodilator response. The results from Cochran Q tests revealed that the frequency of the experience of "chest tightness" decreased significantly across the phases of treatment. However, the sensations of "work" or "breathing effort" persisted at the same time that the FEV1 revealed ongoing airways obstruction. We conclude that attention to the language of dyspnea would alert health care providers to residual air flow obstruction despite decreases in overall dyspnea intensity. PMID- 9731001 TI - Frequencies of T cells expressing interleukin-4 and interleukin-5 in atopic asthmatic children. Comparison with atopic asthmatic adults. AB - T-cell-derived cytokines have been implicated in the pathogenesis of asthma and it has been suggested that Th2-type cytokines (interleukin-4 [IL-4], interleukin 5 [IL-5]) are pivotal in the allergic inflammation. However, there are little data on human cytokine production by individual T cells at the protein level, in particular in asthmatic children. In this study we analyzed the cytokine production at the single cell level in peripheral blood from mild atopic asthmatic (AA) children and adults and age-matched atopic nonasthmatic (AN) and nonatopic nonasthmatic (NN) control subjects (n = 9 in each group) using the technique of intracellular cytokine detection by flow cytometry. Comparing asthmatic children with atopic and nonatopic control subjects, an increased percentage of IL-5-producing T cells (AA: median 4.9% [range 1.1 to 8.9%]; AN: 0.3% [0.2 to 0.9%], p = 0.003; NN: 0.4% [0.1 to 3.8%], p = 0.001) was detectable, with a positive correlation to the number of peripheral eosinophils and to bronchial hyperresponsiveness. The frequency of IL-4-producing T cells was increased in both atopic groups compared with nonatopic controls (AA: 1.2% [0.2 to 2.6%], p = 0.011; AN: 0.8% [0.4 to 3.7%], p = 0.007; NN: 0.4% [0.2 to 0.9%]) with a positive correlation to total IgE concentration. In adults there were no differences in IL-5- or IL-4-producing T cells between all three groups. A substantial proportion of T cells coproducing IL-4 and IL-5 was not detectable in children and adults. These findings indicate that in asthmatic children the frequencies of Th2-type-producing T cells are increased and that expression of IL 4 and IL-5 is regulated independently. PMID- 9731002 TI - Adrenal epinephrine increases alveolar liquid clearance in a canine model of neurogenic pulmonary edema. AB - Case reports of neurogenic pulmonary edema (NPE) often indicate that the edema resolves quickly. Because plasma epinephrine concentration may be elevated in NPE, and epinephrine has been shown to increase the rate of alveolar liquid clearance (ALC), we determined if ALC was increased in a canine model of NPE produced by the intracisternal administration of veratrine. ALC was determined by instilling autologous plasma into a lower lung lobe and using the increase in instillate protein concentration after 4 h to calculate the volume of fluid cleared from the airspaces by mass balance. To prevent pulmonary hypertension and edema, which would confound the mass balance analysis, carotid arterial blood was allowed to drain into a reservoir as pulmonary arterial pressure started to rise after veratrine administration. ALC in animals administered veratrine (n = 6) was 30.4 +/- 1.6 (SE)% of the instilled volume compared with 14.1 +/- 2.1% observed in control animals. The increase in ALC could be inhibited by adrenalectomy, beta2-adrenergic blockade using ICI 118,551, or sodium channel blockade using amiloride and could be duplicated by infusing epinephrine to increase plasma epinephrine concentration to levels observed in NPE. These data indicate that the increased ALC was mediated by adrenal epinephrine and suggest that edema resolution in patients with NPE might be accelerated by endogenous epinephrine. PMID- 9731003 TI - Changes in CD11b and L-selectin expression on eosinophils are mediated by human lung fibroblasts in vitro. AB - Eosinophilic airway infiltration is a central feature in asthma. Eosinophils recovered from bronchoalveolar fluid show an activated phenotype, e.g., increased CD11b and decreased L-selectin expression. We investigated whether lung fibroblasts are able to activate eosinophils in vitro, and if so, which activating factor is most important. CD11b and L-selectin expression of isolated peripheral blood eosinophils were measured by flow cytometry after coculture with normal lung fibroblasts or their conditioned medium. We found that eosinophil CD11b expression increased (154% and 210%, p < 0.05) and L-selectin expression decreased (59% and 35.5%, p < 0.05) on eosinophils compared with baseline (100%) after 4 and 24 h of coculture with interleukin-1-beta (IL-1beta)-stimulated fibroblasts, respectively. Conditioned medium of stimulated fibroblasts also increased CD11b expression, but to a smaller extent (p < 0.05). L-selectin expression of eosinophils in cocultures was not different from that of eosinophils in conditioned medium. Only anti-granulocyte/macrophage colony stimulating factor (anti-GM-CSF) reduced the activation of eosinophils in conditioned medium to almost basal levels (p < 0.05). An increase in CD11b expression is mediated by cytokines as well as direct cell contact, whereas a decrease in L-selectin expression is only mediated by cytokines. GM-CSF released by fibroblasts is an important factor in the modulation of both CD11b and L selectin expression. These results show that lung fibroblasts can activate eosinophils by both adhesive interactions and by soluble factors. PMID- 9731004 TI - Sleep fragmentation indices as predictors of daytime sleepiness and nCPAP response in obstructive sleep apnea. AB - Sleep fragmentation and respiratory disturbance measures are used in the assessment of obstructive sleep apnea (OSA) but have proved to be disappointingly poor correlates of daytime sleepiness. This study investigates the ability of electroencephalograph (EEG) and non-EEG sleep fragmentation indices to predict both presenting sleepiness and the improvement in sleepiness with subsequent nasal continuous positive airway pressure (nCPAP) therapy (nCPAP responsive sleepiness). Forty-one patients (36 men, 5 women), ranging from nonsnorers to severe OSA (> 4% O2 dip rate, median 11.1, range 0.4 to 76.5), had polysomnography with microarousal scoring, computerized EEG analysis, autonomic arousal detection, and body movement analysis. All patients received a trial of nCPAP regardless of sleep study outcome. Spearman's correlation analysis showed significant and similar associations between all sleep fragmentation indices with both pretreatment and nCPAP responsive sleepiness. There was no deterioration in sleepiness on nCPAP in the nonsnorers. Using stepwise multiple regression analysis, the best predictor of nCPAP responsive subjective and objective sleepiness was body movement index, explaining 38% and 43% of the variance, respectively. Variability in EEG sleep depth, quantified from computerized EEG analysis, was the only other index to contribute to these models. Together these indices explained 44% and 51% of the subjective and objective response to nCPAP, respectively. These results suggest that sleep fragmentation indices are useful for identifying OSA patients with sleepiness likely to respond to nCPAP. PMID- 9731005 TI - beta2-Adrenergic receptor haplotypes in mild, moderate and fatal/near fatal asthma. AB - Excess beta2-agonist use in asthmatics has been associated with increased mortality and morbidity. The mechanisms responsible for these observations are unknown. We hypothesized that polymorphisms of the beta2-adrenergic receptor (beta2AR) at amino acid positions 16, 27, and 164, which are known to alter receptor functions in vitro, may predispose asthmatics to fatal/near-fatal asthma and/or modify asthma severity. In preliminary studies we found significant differences in allele frequencies due to ethnic background: Caucasian, Black, Asian Gly16 = 0.61, 0.50, 0.40 and Gln27 = 0.57, 0. 73, 0.80, respectively. beta2AR genotyping was performed on DNA from Caucasians classified as nonasthmatic/nonatopic (n = 84), fatal/near-fatal asthmatics (n = 81) and mild/moderate asthmatics (n = 86). No polymorphism or haplotype was found to be associated with fatal/near-fatal asthma. However, the Gly16/Gln27 haplotype, which undergoes enhanced downregulation in vitro, was substantially more prevalent in moderate asthmatics than in mild asthmatics (p = 0.003, odds ratio = 3.1). We conclude that the beta2AR genotype is not a major determinant of fatal or near-fatal asthma. Furthermore, allele frequency variation among ethnic groups must be considered in clinical studies of beta2AR polymorphisms in asthma. PMID- 9731006 TI - Effects of theophylline on tolerance to the bronchoprotective actions of salmeterol in asthmatics in vivo. AB - Long-term treatment with salmeterol produces tolerance for its protective effects against bronchoconstrictor stimuli in patients with asthma. There is human in vitro evidence that theophylline may prevent beta2-adrenoceptor downregulation. Therefore, we investigated the effect of theophylline on the tolerance to the protective effect of salmeterol against histamine challenge in asthma in vivo. In a parallel 6-wk study, 25 asthmatics were treated with theophylline (mean serum level +/- SEM: 9.9 +/- 1.1 mg/L, Days 1 to 40) or placebo, combined with inhaled salmeterol (50 microgram twice daily, Days 8 to 36). Histamine challenges were carried out by tidal breathing method at entry, and at Days 4, 8, 22, 36, and 40. The response was measured by PC20. There was no significant change in PC20 after 4 d monotherapy with theophylline or placebo (mean difference +/- SEM: 0.54 +/- 0.39 and -0.02 +/- 0.41 doubling dose [DD], respectively; p > 0.15). One hour after the first dose, salmeterol afforded significant protection against histamine, as shown by an increase in PC20 in both the theophylline and placebo group (by 3.49 +/- 0.28 and 3.36 +/- 0.32 DD, respectively; p < 0. 001). However, after 2 and 4 wk salmeterol treatment, the improvements in PC20 by salmeterol were significantly reduced to 1. 80 +/- 0.35 and 1.69 +/- 0.36 DD, respectively, in the theophylline group (p < 0.001), and to 1.55 +/- 0.47 and 1.52 +/- 0.56 DD, respectively, in the placebo group (p < 0.002). These changes were not significantly different between the groups (p > 0.80). After cessation of salmeterol treatment, PC20 was not significantly different from the values at entry in either group (p > 0.90). We conclude that regular theophylline treatment neither prevents, nor worsens, the development of tolerance to the bronchoprotective effect of salmeterol in asthmatics in vivo. PMID- 9731007 TI - Plasma and muscle amino acid levels in relation to resting energy expenditure and inflammation in stable chronic obstructive pulmonary disease. AB - In patients with chronic obstructive pulmonary disease (COPD), muscle wasting can occur independently of fat loss, suggesting disturbances in protein metabolism. In order to provide more insight in amino-acid (AA) metabolism in patients with stable COPD, we examined arterial plasma and anterior tibialis muscle AA levels, comparing 12 COPD patients with eight age-matched healthy control subjects. We also studied relationships between AA levels, the acute phase response as measured by lipopolysaccharide-binding protein (LBP), and resting energy expenditure (REE). In contrast to findings in acute diseases associated with muscle wasting, we found increased muscle glutamine (GLN) levels in our patient group (mean +/- SEM = 10,782 +/- 770 versus 7,844 +/- 293 micromol/kg wet weight, p < 0. 01). Furthermore, muscle arginine, ornithine, and citrulline were significantly increased in the patient group, whereas glutamic acid was decreased. In plasma, the sum of all AA (SumAA) was decreased in the patient group (2,595 +/- 65 versus 2,894 +/- 66 micromol/L, p < 0.01), largely because of decreased levels of alanine (254 +/- 10 versus 375 +/- 25 micromol/L, p < 0.0001), GLN (580 +/- 17 versus 641 +/- 17 micromol/L, p < 0.05), and glutamic acid (91 +/- 5 versus 130 +/- 10 micromol/L, p < 0.01). LBP levels were increased in COPD patients as compared with controls (11.7 +/- 4.5 versus 8.6 +/- 1.0 mg/L, p < 0.05), and showed a positive correlation with REE (r = 0. 49, p = 0.03), a negative correlation with the SumAA in plasma (r = -0.76, p < 0.0001), and no correlation with muscle AA levels. In conclusion, various disturbances in plasma and muscle AA levels were found in COPD patients. A relationship between the observed decreased plasma AA levels and inflammation was suggested. PMID- 9731008 TI - Maternal cigarette smoking is associated with increased inner airway wall thickness in children who die from sudden infant death syndrome. AB - The harmful effects of passive cigarette smoke exposure to infants include an increased frequency of asthma exacerbations, lower respiratory viral infections, and the sudden infant death syndrome (SIDS). Because of a difficulty in obtaining airway tissue from infants, little information is available on the effects of passive cigarette smoke exposure on the structure of the infant airway wall. We examined airway dimensions in 19 children who died from SIDS whose mothers smoked more than 20 cigarettes a day prenatally and postnatally, and compared these data with those from 19 infants who died from SIDS and had nonsmoking mothers. Total inner and outer wall areas were calculated for each airway and expressed in terms of the basement membrane perimeter (Pbm). Inner airway wall thickness was greater in the larger airways of those infants whose mothers had smoked more than 20 cigarettes a day. These findings suggest that infants exposed to a high level of passive cigarette smoke develop significant structural changes in their airways. Increased airway wall thickness may contribute to exaggerated airway narrowing and may help explain the previously observed abnormalities in neonatal lung function that have been described in infants of smoking mothers. PMID- 9731009 TI - Radiographic evidence of silicosis risk in the diatomaceous earth industry. AB - There is limited and conflicting evidence regarding the exposure-response relationship between exposure to crystalline silica and silicosis; the level of risk to current workers remains uncertain. We conducted an epidemiologic investigation of 1,809 workers in the diatomaceous earth industry, where exposures to crystalline silica are primarily to the cristobalite form. On the basis of the median of three independent readings, 81 (4.5%) workers were judged to have opacities on chest radiographs (small opacities, profusion >= 1/0, and/or large opacities). Age-adjusted relative risk of opacities increased significantly with cumulative exposure to crystalline silica. The concentration of respirable crystalline silica to which workers were exposed (highly correlated with period of hire) was an important determinant of risk after accounting for cumulative exposure. For workers with an average exposure to crystalline silica of <= 0.50 mg/m3 (or hired >= 1950), the cumulative risk of opacities for a cumulative exposure to crystalline silica of 2.0 mg/m3-yr was approximately 1.1%; for an average exposure > 0.50 mg/m3 (or hired < 1950), the corresponding cumulative risk was 3.7%. These findings indicate an exposure-response relationship between cumulative exposure to crystalline silica and radiographic opacities; moreover, the relationship was substantially steeper among workers exposed at the highest average concentrations of crystalline silica. PMID- 9731010 TI - Pseudophysiologic emphysema resulting from severe small-airways disease. AB - Loss of lung elastic recoil causing hyperinflation with increased TLC and decreased diffusing capacity and expiratory airflow are physiologic hallmarks of emphysema. We studied lung mechanics in 10 patients (seven men and three women) aged 69 +/- 9 yr (mean +/- SD) who had fixed, severe expiratory airflow limitation with a mean FEV1 = 0.73 +/- 0.1 L (mean +/- SD) (32 +/- 7% predicted) and lung computed tomographic picture grade score <= 20, indicating no or trivial emphysema. Three patients died, in whom whole-lung emphysema scores were 15 each and small airways were abnormal. Marked hyperinflation was present in all 10 patients studied, with TLC 7.3 +/- 1.1 L (140 +/- 12% predicted); FRC 5.6 +/- 0.8 L (177 +/- 30% predicted); and RV 5.2 +/- 0.8 L (242 +/- 28% predicted). Diffusing capacity of carbon monoxide (DLCO was reduced, at 12 +/- 6 ml/min/mm Hg (61 +/- 29% predicted). The pressure-volume curves of the lung were markedly abnormal. Pst(L) at TLC was 11.6 +/- 1.4 cm H2O. Transdiaphragmatic pressure (Pdi) in five patients was 66 +/- 13 cm H2O. These results indicate that severe small-airways disease with no or trivial emphysema may cause a spurious reduction in diffusing capacity as well as severe loss of lung elastic recoil resulting in marked hyperinflation, increased TLC, and decreased Pdi and expiratory airflow. PMID- 9731011 TI - Electrophysiologic and inotropic effects of K+-channel blockade in aged diaphragm. AB - The aminopyridines block several types of potassium (K+) channels and exert a direct inotropic effect on skeletal muscle by prolonging the duration of the action potential. Aging influences skeletal muscle Cl- channels and their regulation, and affects both resting whole-cell K+ conductance and adenosine triphosphate (ATP)-sensitive K+ channels, although in opposite directions. The present study tested the hypothesis that aging affects diaphragm-muscle K+ channels responsible for repolarization of the action potential and force production. Diaphragms of young adult (age 3 to 4 mo) and old (age 20 to 21 mo) male Fischer 344 rats was studied in vitro at 37 degrees C. The K+-channel blocker 3,4-diaminopyridine (DAP, 0.3 mM) did not alter resting membrane potential or action-potential height, overshoot, or rate of depolarization of either young-adult or old muscle. However, DAP slowed the rate of repolarization of the action potential and increased the action-potential area in young-adult and old muscle; the time for the action potential to repolarize by 80% increased from 0.59 +/- 0.02 ms (mean +/- SE) to 3.37 +/- 0.68 ms (p < 0.05) in young-adult muscle and from 0.87 +/- 0.06 ms to 2.52 +/- 0.54 ms (p < 0.05) in old muscle, whereas the action-potential area increased from 56 +/- 3 mVms to 193 +/- 34 mVms (p < 0.05) in young-adult muscle and from 72 +/- 5 mVms to 134 +/- 20 mVms (p < 0. 05) in old muscle. The action-potential area was not different in young-adult and old diaphragm without DAP, but was significantly larger in young-adult than in old diaphragm with DAP (p < 0.05). The functional consequence was that DAP increased diaphragm isometric twitch force by 181 +/- 12% (p < 0.05) in young adult muscle and by 144 +/- 24% (p < 0.05) in old muscle; the increase was significantly greater in young-adult than in old muscle (p < 0.05). These data suggest an aging-associated reduction in, or reduced DAP sensitivity of, diaphragm K+ conductance during action potentials, which most likely reflects aging-associated alterations in delayed-rectifier K+ conductance. Although the inotropic effect of DAP was greater for young-adult than for old diaphragm muscle, the difference was sufficiently modest to show that DAP has substantial inotropic effects in old muscle. PMID- 9731012 TI - Comparison of serial monitoring of peak expiratory flow and FEV1 in the diagnosis of occupational asthma. AB - Peak expiratory flow (PEF) monitoring is often used to establish the relationship between occupational exposure and asthma. FEV1 has been found to be a better physiologic index than PEF in the measurement of airflow obstruction. The aim of this study was to compare the accuracy of serial monitoring of PEF and FEV1 in the diagnosis of occupational asthma. Twenty consecutive subjects referred for possible occupational asthma were asked to perform serial monitoring of PEF and FEV1 using a portable ventilometer. Two sets of graphs were plotted for both PEF and FEV1: graphs with the best of all values and graphs with the best of two reproducible values. Three observers interpreted both PEF and FEV1 recordings by the visual method in a blind, randomized manner as either compatible with occupational asthma or not. Eleven of the subjects had a positive inhalation challenge test (high-molecular-weight agents, n = 6; low-molecular-weight agents, n = 5). In the case of analysis of the graphs plotted with the best of all values, the sensitivity of the PEF recording interpreted by the three observers was 82, 73, and 73%, and of the FEV1 recording as 55, 55, and 45%; specificity of PEF recording was 89, 100, and 100%, and of FEV1 was 56, 89, and 100%. When an agreement between two of the three readers was required to define occupational asthma, sensitivity and specificity were 73 and 100% for PEF and 55 and 89% for FEV1. Lower sensitivities were found when the same analyses were performed with the graphs plotted with the best of two reproducible values. It was concluded that unsupervised FEV1 is not more accurate than unsupervised PEF monitoring in the diagnosis of occupational asthma. Plotting graphs using the best value gives better diagnostic accuracy than plotting them with the best of two reproducible values. PMID- 9731013 TI - Impact of inhaled nitric oxide on platelet aggregation and fibrinolysis in rats with endotoxic lung injury. Role of cyclic guanosine 5'-monophosphate. AB - As inhaled nitric oxide (iNO) may differently increase bleeding time (BT) and inhibit platelet aggregation in normal and lung-injured patients or experimental models, we studied the effects of iNO on hemostasis in presence and absence of an endotoxic lung injury in the rat. Eight hours after intratracheal administration of endotoxin (lipopolysaccharide [LPS]) or its solvent (phosphate-buffered solution [PBS]), four groups of rats were randomized according to the presence or absence of 15 ppm iNO added for an additional 10 h. We measured BT, ex vivo platelet aggregation, plasma fibrinogen, euglobulin clot lysis time (ECLT), and platelet and aortic cyclic guanosine 5'-monophosphate (cGMP) contents. Acute lung inflammation did not influence BT, but increased platelet aggregability, fibrinogen levels, and platelet and aortic cGMP. In control and endotoxic rats, iNO increased BT, reduced platelet aggregability, and increased platelet cGMP. iNO increased aortic cGMP only in healthy rats. ECLT was increased by LPS and unchanged with iNO. These results suggest that the extrapulmonary "systemic" effects induced by iNO on hemostasis were not strictly similar in healthy and LPS rats, inflammation inducing proper changes in coagulation parameters. However, iNO attenuated the procoagulant activity induced by acute lung inflammation, suggesting a potentially beneficial effect of this therapy. PMID- 9731014 TI - Aerosolized surfactant improves pulmonary function in endotoxin-induced lung injury. AB - Surfactant dysfunction is a primary pathophysiologic component in patients with adult respiratory distress syndrome (ARDS). In this study we tested the efficacy of aerosolized surfactant (Sf ) replacement in a severe lung injury model of endotoxin-induced ARDS. Twenty-one certified healthy pigs were anesthetized, surgically prepared for measurement of hemodynamic and lung function, then randomized into one of four groups: (1) control (n = 5), surgical instrumentation only; (2) lipopolysaccharide (LPS) (n = 6), infused with Escherichia coli LPS (100 microgram/kg) without positive end- expiratory pressure (PEEP) and ventilated with a nonhumidified gas mixture of 50% N2O and 50% O2; (3) LPS + PEEP (n = 4), infused with LPS, placed on PEEP (7.5 cm H2O), and ventilated with a humidified gas mixture; and (4) LPS + PEEP + Sf (n = 6), infused with LPS, placed on PEEP, and ventilated with aerosolized Sf (Infasurf, ONY, Inc.). All animals were studied for 6 h. Arterial PO2 significantly decreased in both the LPS and LPS + PEEP groups (LPS + PEEP = 74 +/- 19 mm Hg; LPS = 74 +/- 19 mm Hg, p < 0.05) while venous admixture (Q S/Q T) increased in these groups (LPS + PEEP = 43.3 +/- 3.9%; LPS = 47.7 +/- 11%, p < 0.05) as compared with the control group. PEEP + Sf reduced the fall in PO2 (142 +/- 20 mm Hg) and rise in Q S/Q T (15.1 +/- 3.6%) caused by LPS. Delayed induction of PEEP (2 h following LPS) did not significantly improve any parameter over the LPS group without PEEP in this ARDS model. LPS without PEEP (3.4 +/- 0.2 cells/6,400 micrometer2) caused a marked increase in the total number of sequestered leukocytes in the pulmonary parenchyma as compared with the control group (1.3 +/- 0.1 cells/6,400 micrometer2). LPS + PEEP + Sf (2.3 +/- 0.2 cells/6,400 micrometer2) significantly decreased while LPS + PEEP significantly increased (4.0 +/- 0.2 cells/6,400 micrometer2) the total number of sequestered leukocytes as compared with the LPS without PEEP group. In summary, aerosolized surfactant replacement decreased leukocyte sequestration and improved oxygenation in our porcine model of endotoxin-induced lung injury. PMID- 9731016 TI - Sex differences in mortality of people who visited emergency rooms for asthma and chronic obstructive pulmonary disease. AB - We assess the sex differences in mortality in a population-based cohort of those Barcelona residents older than 14 yr of age who received emergency room services (ERS) for either chronic obstructive pulmonary disease (COPD) or asthma, during the period from 1985 to 1989. Vital status was followed to the end of 1995. A total of 15,517 individuals, 9,918 males and 5,599 females were included in the study. Asthma was diagnosed in 16% of males and 53% of females. Overall, 50% of males and 30% of females died during the follow-up period. The mortality rates in both males and females who visited emergency rooms for COPD or asthma were significantly higher than the expected rates in the general population. These relative increases in the mortality rates were significantly higher in females than in males for both causes of death, COPD (age-adjusted female/male ratio = 2.39), and asthma (ratio = 3.95). However, survival was better in females than males among individuals in the study. The higher fatality in males than females was observed for all causes of death, all respiratory causes, and COPD (risk ratio among patients with COPD = 0.42, 0.29-0.59, and among patients with asthma = 0.11, 0.02-0.60), but not for asthma. Mortality for asthma was higher in females with a diagnosis of COPD (2.79, 1.52-5.13), but it was not different among individuals in whom asthma was diagnosed (1.02, 0.56-1.87). Greater severity of COPD in males than in females could explain a higher risk of dying for all respiratory causes and COPD in males. The increased risk of asthma death in females may be due to problems of coding the term "asthma" in death certificates. The higher rates in females than in males when comparing with the general population, may be an expression of a greater similarity in risk factors, such as smoking, in our population than in males and females of the general population. PMID- 9731015 TI - Contribution of HLA genes to genetic predisposition in diffuse panbronchiolitis. AB - Diffuse panbronchiolitis (DPB) in East Asia is a distinctive chronic obstructive pulmonary disease of unknown etiology. We hypothesize that the disease susceptibility is due to genetic predisposition unique to Asians. Association between human leukocyte antigen (HLA)-Bw54 and the disease was previously reported. In the present study, using newly developed polymerase chain reaction (PCR)- based methods, we directly analyzed HLA class I and II alleles in 76 Japanese patients. HLA-A, -B, and -C antigens were screened by the conventional typing method, and then B22-group alleles including HLA-B54 were genotyped by single-strand conformation polymorphism analysis. Alleles of HLA-DRB1 gene were fully determined by the microtiter plate hybridization method. Thirty-seven percent of the patients possessed HLA-B*5401 allele conserved predominantly in East Asians, as compared with 15% of 110 healthy volunteers (chi2 = 12.4, p = 0.0004). In addition, 4% of the patients possessed B*5504 also unique to Asians but a rare allele which was not found in normal control subjects in this study. Typing of HLA-DRB1 class II gene did not demonstrate strong positive association with the disease. A33, B44, and DRB1*1302 showed negative association with the disease. We conclude that distinctive molecular structure of HLA-B alleles or a closely linked gene in the HLA region contributes to genetic predisposition in diffuse panbronchiolitis. This may partly explain why this disorder is found primarily in Asians. PMID- 9731017 TI - Risk of emergency care, hospitalization, and ICU stays for acute asthma among recipients of salmeterol. AB - We used automated health insurance claims records of a New England insurer to assess the relation between salmeterol and severe nonfatal asthma. We identified 61,712 members who received a beta-agonist from January 1, 1993 to August 31, 1995, including 2, 708 recipients of salmeterol. Compared with recipients of other beta-agonists, future salmeterol recipients had higher rates of asthma hospitalization and dispensings of asthma medications during the year before they received salmeterol. We selected as a comparison group 3,825 recipients of sustained-release theophylline. We defined a baseline period as the year before the start of the follow-up period, and we characterized patients according to age, sex, calendar period, presence of baseline hospitalizations for asthma, presence of chronic obstructive pulmonary disease (COPD), and baseline dispensings of asthma medications. After adjusting for baseline factors, incidence rates of severe asthma in the salmeterol group were not elevated for emergency care (rate ratio estimate [RR] = 0.69, 95% confidence intervals [CI] = 0.42, 1.11), hospitalization (RR = 1.09, 95% CI = 0.60, 1.98), or intensive care unit (ICU) stays (RR = 0.81, 95% CI = 0.25, 2.62). We conclude that salmeterol was prescribed preferentially to high-risk patients and, after adjusting for baseline risk, salmeterol recipients did not have a greater risk than theophylline recipients of severe nonfatal asthma. PMID- 9731018 TI - Utility of a provocation test for diagnosis of chronic pigeon Breeder's disease. AB - Chronic hypersensitivity pneumonitis (CHP) can be difficult to differentiate from other interstitial lung diseases (ILD). To determine the diagnostic usefulness of a provocation test (PT), 17 patients with CHP induced by avian antigens, 17 with other ILD, and five healthy control subjects were challenged with pigeon serum. After PT, an increase in body temperature (BT) and a decrease in FVC, PaO2 and SaO2% were observed in all patients with CHP and in three with ILD. No reaction was noticed in healthy subjects. ROC curves showed that for FVC the best cut point was a drop of 16% displaying sensitivity (S): 76%, specificity (SP): 81%, positive predictive value (PPV): 81%, and negative predictive value (NPV): 83%. For a drop of 3 mm Hg in PaO2 or 3% SaO2, S was 88% for both, SP was 82 and 86%, PPV was 81 and 82%, and NPV was 82 and 86%, respectively. An increase of BT > 0.5(o) C showed S, 100%; SP, 82%; PPV, 100%; NPV, 86%. A univariate regression analysis confirmed that changes in BT and FVC are predicting values of CHP: RR, 82.5 (CI, 10.43 to 651.76) and 1.21 (CI, 1.06 to 1.36). There were no challenge test complications. These findings suggest that PT is a useful tool for diagnosis of CHP. PMID- 9731019 TI - Pneumonia in acute respiratory distress syndrome. A prospective evaluation of bilateral bronchoscopic sampling. AB - We evaluated the diagnostic yield of bilateral bronchoalveolar lavage (BAL) in patients with acute respiratory distress syndrome (ARDS) with suspected ventilator-associated pneumonia (VAP) and compared BAL results from contralateral sites. Ninety-four ARDS patients with suspected VAP underwent 172 bronchoscopies (344 BALs). BAL was processed for quantitative cultures, total cell count and subjected to microscopic analysis for cell differential, presence of intracellular organisms (ICO), and Gram stain. The diagnostic threshold for VAP was a growth of >= 10(4) cfu/ml in BAL culture. Most episodes (68%) had bilateral insignificant bacterial growth. Forty (43%) patients had one or more episodes of VAP. Thirty-three of the 55 (60%) positive bronchoscopies had significant growth in only one side, 18 were right BAL, and 15 were left BAL. Episodes with bilateral significant growth were more likely to be polymicrobial, to have a bacterial growth >= 10(5) cfu/ml in the BAL, and to possess a higher percentage of neutrophils and ICO. Among 65 microorganisms recovered in significant concentration, Pseudomonas aeruginosa occurred in 43% and S. aureus in 15%. Overall, Gram stain had a sensitivity of 54% and a specificity of 87%; and Giemsa stain (> 2% ICO) had a sensitivity of 46% and a specificity of 93%. Antibiotic treatment was modified by the results of BAL cultures in 50 (91%) episodes of pneumonia. In patients with ARDS and suspected VAP, bilateral BAL quantitative bacterial cultures had significant growth on one side only in 19% and in both sides in 13%. PMID- 9731020 TI - Prognostic indicators for blood and marrow transplant patients admitted to an intensive care unit. AB - Although hematopoietic stem cell transplantation (HSCT) can be curative in patients with certain malignancies, survival is poor if the recipient becomes critically ill. This prospective study examined the outcomes of 115 consecutive HSCT patients admitted to the medical intensive care unit (MICU) of a tertiary cancer center and identified variables associated with survival. The need for endotracheal intubation and mechanical ventilation ("intubation") had a profound adverse effect on survival. Overall, 9 of 48 (18.8%) intubated patients survived compared with a survival rate of 44 of 67 (65.7%) among patients not intubated (p < 0.001). This pattern persisted for nearly all patient subgroups. Among intubated patients, those receiving peripheral blood stem cell transplant (PBSCT) had significantly better survival than bone marrow transplant (BMT) patients (8 of 26, 31% versus 1 of 22, 4%; p = 0.028). Multiple logistic regression analyses indicated that the probability a patient admitted to the MICU survived decreased significantly if the patient was intubated, had an allogeneic rather than autologous transplant, had an infection or gastrointestinal bleeding, and also decreased with higher respiratory rate, higher heart rate, longer time from transplant to MICU admission or higher bilirubin. These results may be of value in deciding which critically ill patients will benefit from intubation following major complications after HSCT transplantation. PMID- 9731021 TI - Lung resection for invasive pulmonary aspergillosis in neutropenic patients with hematologic diseases. AB - Invasive pulmonary aspergillosis (IPA) is associated with a high mortality. In 27 consecutive neutropenic patients who underwent lung resection for suspected IPA, we analyzed preoperative diagnostic evaluation, operative procedure, perioperative management, histological findings, outcome concerning recurrence of aspergillosis, and survival to evaluate the morbidity and mortality of a surgical treatment of IPA. Seventeen patients with hematologic diseases had previously undergone high-dose chemotherapy and four stem cell transplantation. Six patients with aplastic anemia were treated with antilymphocyte globulin. IPA was suspected if localized infiltrates developed on thoracic CT scan, and fever persisted under antibiotic therapy in neutropenic patients. In only one case a diagnosis of IPA could be made preoperatively. Twenty patients underwent lobectomy and seven wedge resection. At day of surgery the neutrophil count was below 500 x 10(9)/L in 78% of patients, and the platelet count below in 50 x 10(9)/L in 58% of patients. Invasive fungal infection was confirmed histologically in 22 of 27 patients (81.5%); in five patients no fungal infection was documented. The median duration of surgery was 120 min. Postoperatively, patients stayed one night in the intensive care unit, and chest tubes were removed after 2 d. Within 7 d a median of four erythrocyte packs and two platelet packs per patient were replaced. Major surgical complications occurred in two patients (bronchial dehiscence; pleural aspergillosis). Minor surgical complications included prolonged chest tube drainage (recurrent pneumothorax, n = 2; air leakage, n = 1; hematothorax, n = 1), pleural effusion (n = 4), and seroma (n = 2). Postoperatively, two patients suffered from histologically proven disseminated aspergillosis (pleural aspergillosis, renal aspergilloma) and another patient from suspected orbital aspergillosis. At 30 d postoperative mortality was 11% and 3-mo survival was 77%. After lung resection, seven patients underwent stem cell transplantation without recurrence of IPA. In conclusion, we suggest lung resection is a therapeutic option for invasive pulmonary aspergillosis in neutropenic patients with hematologic diseases and is associated with a low surgery-related morbidity and mortality. PMID- 9731022 TI - Respiratory symptoms in children and indoor exposure to nitrogen dioxide and gas stoves. AB - Nitrogen dioxide levels were measured in 80 homes in the Latrobe Valley, Victoria, Australia, using passive samplers. Some 148 children between 7 and 14 yr of age were recruited as study participants, 53 of whom had asthma. Health outcomes for the children were studied using a respiratory questionnaire, skin prick tests, and peak flow measurements. Nitrogen dioxide concentrations were low, with an indoor median of 11.6 microgram/m3 (6.0 ppb), and a maximum of 246 microgram/m3 (128 ppb). Respiratory symptoms were more common in children exposed to a gas stove (odds ratio 2.3 [95% CI 1. 0-5.2], adjusted for parental allergy, parental asthma, and sex). Nitrogen dioxide exposure was a marginal risk factor for respiratory symptoms, with a dose-response association present (p = 0.09). Gas stove exposure was a significant risk factor for respiratory symptoms even after adjusting for nitrogen dioxide levels (odds ratio 2.2 [1.0-4.8]), suggesting an additional risk apart from the average nitrogen dioxide exposure associated with gas stove use. Atopic children tended to have a greater risk of respiratory symptoms compared with nonatopic children with exposure to gas stoves or nitrogen dioxide, but the difference was not significant. PMID- 9731023 TI - Nasal potential difference in congenital bilateral absence of the vas deferens. AB - Congenital bilateral absence of the vas deferens (CBAVD) is supposed to be due to defective activity of the cystic fibrosis (CF) transmembrane conductance regulator (CFTR) in the genital tract. With the aim of studying CFTR activity in vivo we measured nasal potential difference (NPD) in a group of CBAVD subjects, who were then compared with normal control subjects and CF patients. Sodium transport, measured under basal conditions and after amiloride superinfusion, was normal in almost all CBAVD patients, who had NPD values similar to those of normal control subjects. Chloride transport was studied by measuring NPD during perfusion with a chloride-free solution and isoproterenol. Under these circumstances CBAVD patients as a whole showed normal chloride secretion. However, three subjects with CBAVD had abnormal NPD values. They had either elevated sweat chloride concentrations together with symptoms of mild CF, or compound heterozygosity (DeltaF508/R117H). In conclusion the group of CBAVD patients as a whole presented normal bioelectric properties of nasal epithelium, suggesting normal CFTR activity. In a small subgroup NPD was abnormal, suggesting a diagnosis of CF, later confirmed by elevated sweat chloride concentrations or positive DNA testing. We suggest that CBAVD patients with altered NPD should undergo further clinical follow-up in order to detect possible late complications of CF. PMID- 9731024 TI - Exhaled nitric oxide is higher both at day and night in subjects with nocturnal asthma. AB - Nitric oxide in exhaled air is thought to reflect airway inflammation. No data have been reported so far on circadian changes in NO in subjects with nocturnal asthma. To determine whether exhaled NO shows a circadian rhythm inverse to the circadian rhythm in airway obstruction in subjects with nocturnal asthma, we conducted a study involving six healthy controls, eight individuals without nocturnal asthma (4-h to 16-h variation in peak expiratory flow [PEF] <= 15%), and six individuals with nocturnal asthma (4-h to 16-h PEF variation > 15%). Smoking, use of corticosteroids, and recent respiratory infections were excluded. NO concentrations were measured at 12, 16, 20, and 24 h, and at 4, 8, and 12 h of the next day, using the single-breath method. At the same times, FEV1 and PEF were also measured. Mean NO concentrations were significantly higher in subjects with nocturnal asthma than in subjects without nocturnal asthma, and higher in both groups than in healthy controls at all time points. Mean exhaled NO levels over 24 h correlated with the 4-h to 16-h variation in PEF (r = 0.61, p < 0.01). Exhaled NO did not show a significant circadian variation in any of the three groups as assessed with cosinor analysis, in contrast to the FEV1 in both asthma groups (p < 0.05). At 4 h, mean +/- SD NO levels were higher than at 16 h in subjects with nocturnal asthma; at 50 +/- 20 ppb versus 42 +/- 15 ppb (p < 0.05); other measurements at all time points were similar. Differences in NO and FEV1 from 4 h to 16 h did not correlate with one another. We conclude that subjects with nocturnal asthma exhale NO at higher levels both at night and during the day, which may reflect more severe diurnal airway-wall inflammation. A circadian rhythm in exhaled NO was not observed. NO levels did not correspond to the circadian rhythm in airway obstruction. The small increase in NO at 4 h may indicate an aspect of inflammation, but it is not associated with increased nocturnal airway obstruction. PMID- 9731025 TI - Effectiveness and cost of selective decontamination of the digestive tract in critically ill intubated patients. A randomized, double-blind, placebo controlled, multicenter trial. AB - We evaluated the effect of selective decontamination of the digestive tract (SDD) on the incidence of ventilator-associated pneumonia (VAP) and its associated morbidity and cost in a mixed population of intubated patients. Two hundred seventy-one consecutive patients admitted to the intensive care units (ICUs) of five teaching hospitals and who had an expected need for intubation exceeding 48 h were enrolled and received topical antibiotics or placebo. Uninfected patients additionally received ceftriaxone or placebo for 3 d. VAP occurred in 11.4% of SDD-treated and 29.3% of control-group patients (p < 0.001; 95% confidence interval [CI]: 7.8 to 27.9). The incidence of nonrespiratory infections in the two groups was 19.1% and 30.7%, respectively (p = 0.04; 95% CI: 0.7 to 22.7). Among survivors, the median length of ICU stay was 11 d (interquartile range: 7 to 21.5 d) for the SDD-treated group and 16. 5 d (10 to 30 d) for the control group (p = 0.006). Mean cost per survivor was $11,926 for treated and $16,296 for control-group patients. Mortality was 38.9% and 47.1%, respectively (p = 0.57). In decontaminated patients, the prevalence of gram-negative bacilli fell within 7 d from 47.4% to 13.0% (p < 0.001), whereas colonization with resistant gram positive strains was higher (p < 0. 05) than in the placebo group. In a mixed population of intubated patients, SDD was associated with a significant reduction of morbidity at a reduced cost. Our findings support the use of SDD in this high risk group. PMID- 9731026 TI - Biochemical reaction products of nitric oxide as quantitative markers of primary pulmonary hypertension. AB - Primary pulmonary hypertension (PPH) is a rare and fatal disease of unknown etiology. Inflammatory oxidant mechanisms and deficiency in nitric oxide (NO) have been implicated in the pathogenesis of pulmonary hypertension. In order to investigate abnormalities in oxidants and antioxidants in PPH, we studied intrapulmonary NO levels, biochemical reaction products of NO, and antioxidants (glutathione [GSH], glutathione peroxidase [GPx], and superoxide dismutase [SOD]) in patients with PPH (n = 8) and healthy controls (n = 8). Intrapulmonary gases and fluids were sampled at bronchoscopy. Pulmonary hypertension was determined by right-heart catheterization. NO and biochemical reaction products of NO in the lung were decreased in PPH patients in comparison with healthy controls (NO [ppb] in airway gases: control, 8 +/- 1; PPH, 2.8 +/- 0. 9; p = 0.016; and NO products [microM] in bronchoalveolar lavage fluid [BALF]: control, 3.3 +/- 1.05; PPH, 0.69 +/- 0.21; p = 0.03). However, GSH in the lungs of PPH patients was higher than in those of controls (GSH [microM] in BALF: 0.55 +/- 0.04; PPH, 0.9 +/- 0.1; p = 0.015). SOD and GPx activities were similar in the two groups (p >/= 0.50). Biochemical reaction products of NO were inversely correlated with pulmonary artery pressures (R = -0.713; p = 0.047) and with years since diagnosis of PPH (R = -0.776; p = 0.023). NO reaction products are formed through interactions between oxidants and NO, with the end products of reaction dependent upon the relative levels of the two types of molecules. The findings of the study therefore show that NO and oxidant reactions in the lung are related to the increased pulmonary artery pressures in PPH. PMID- 9731027 TI - Potential masking effects of salmeterol on airway inflammation in asthma. AB - We hypothesized that regular use of long-acting beta-agonists could delay recognition of ("mask") increasing airway inflammation. We studied steroid sparing and "masking" effects of salmeterol versus placebo in 13 asthmatic individuals requiring >= 1,500 microgram inhaled corticosteroid daily. Corticosteroid doses were reduced weekly until criteria were met for an exacerbation or the corticosteroid was fully withdrawn. Subjects were restabilized on their original dose of inhaled corticosteroid for 4 wk before crossover to the alternative treatment. Subjects maintained symptom and peak expiratory flow (PEF) diaries, and underwent weekly spirometric, methacholine challenge, sputum eosinophil, and serum eosinophil cationic protein (ECP) measurements. Mean corticosteroid dose was reduced by 87% during salmeterol treatment, versus 69% with placebo (p = 0.04). Sputum eosinophils increased before exacerbation despite stable symptoms, FEV1, and PEF. In the week before clinical exacerbation, sputum eosinophil counts were higher in the salmeterol treatment arm (19.9 +/- 29.8% [mean +/- SD], versus placebo 9.3 +/- 17.6%; p = 0.006). Five subjects showed > 10% sputum eosinophilia before exacerbation during salmeterol treatment, as compared with two receiving placebo. In this model, salmeterol controlled symptoms and lung function until inflammation became significantly more advanced. We conclude that the bronchodilating and symptom relieving effects of salmeterol can mask increasing inflammation and delay awareness of worsening asthma. PMID- 9731029 TI - Angiotensin-converting enzyme inhibition delays pulmonary vascular neointimal formation. AB - Primary pulmonary hypertension (PPH) is a disease characterized pathologically by pulmonary artery medial hypertrophy, adventitial thickening, and neointimal proliferation. Increasing recognition of the importance of remodeling to the pathogenesis of PPH suggests new therapeutic possibilities, but it will be necessary to (1) identify essential mediators of remodeling, and (2) demonstrate that inhibiting those mediators suppresses remodeling before new antiremodeling therapies can be considered feasible. The effect of angiotensin-converting enzyme (ACE) inhibition on pulmonary vascular remodeling was studied in a newly developed rat model in which neointimal lesions develop between 3 and 5 wk after monocrotaline injury is coupled with increased pulmonary artery blood flow after contralateral pneumonectomy. Neointimal formation was significantly suppressed at 5 wk by ACE inhibition whether it was started 10 d before or 3 wk after remodeling was initiated, although medial hypertrophy and adventitial thickening still developed. By 11 wk, the extent of neointimal formation in rats treated with ACE inhibition was similar to rats without ACE inhibition at 5 wk. Pulmonary artery pressures and right ventricular weights correlated with the extent of neointimal formation. Northern blot analysis and in situ hybridization demonstrated marked suppression of lung tropoelastin and type I procollagen gene expression in the presence of ACE inhibition. An angiotensin II type I receptor antagonist partially, but not completely, replicated the effects of ACE inhibition. These data suggest that the tissue angiotensin system may be a target for therapeutic efforts to suppress the vascular remodeling that is characteristic of primary pulmonary hypertension. PMID- 9731028 TI - Inhaled nitric oxide primes lung macrophages to produce reactive oxygen and nitrogen intermediates. AB - Inhaled nitric oxide is a selective pulmonary vasodilator used for the treatment of pulmonary hypertension. The potential adverse effects of inhaled nitric oxide are unknown and represent the focus of the present studies. Whereas inhalation of nitric oxide (10 to 100 ppm, 5 h) by Balb/c mice had no effect on the number or type of cells recovered from the lung, a dose-related increase in bronchoalveolar lavage protein was observed, suggesting that nitric oxide induces alveolar epithelial injury. To determine if this was associated with altered alveolar macrophage activity, we quantified production of reactive oxygen and nitrogen intermediates by these cells. Interferon-gamma, alone or in combination with lipopolysaccharide (LPS), induced expression of inducible nitric oxide synthase (iNOS) protein and nitric oxide production by alveolar macrophages. Cells from mice exposed to 20 to 100 ppm nitric oxide produced significantly more nitric oxide and expressed greater quantities of iNOS than cells from control animals. Superoxide anion production and peroxynitrite generation by alveolar macrophages were also increased after exposure of mice to nitric oxide. This was correlated with increased antinitrotyrosine antibody binding to macrophages in histologic sections. Taken together, these data demonstrate that inhaled nitric oxide primes lung macrophages to release reactive oxygen and nitrogen intermediates. Increased production of these mediators by macrophages following inhalation of nitric oxide may contribute to tissue injury. PMID- 9731030 TI - Interleukin-5 expression in the bone marrow of sensitized Balb/c mice after allergen challenge. AB - Interleukin-5 (IL-5) is a potent eosinophilopoietic factor implicated in the chronic inflammatory cell accumulation accompanying bronchial asthma. However, its role in stimulating eosinophil differentiation within the bone marrow following allergen exposure remains to be elucidated. The aims of our study were to determine the expression of IL-5 within the bone marrow of sensitized and control mice after allergen exposure, and to investigate the cellular phenotype of IL-5-producing cells. Sensitized Balb/c mice were challenged with either ovalbumin (OVA) or sterile saline. After 6 h, the mice were exsanguinated and the bone marrow prepared for cytospins. Bone marrow-derived cells from OVA-sensitized mice exhibited an increase in IL-5 immunoreactivity and mRNA compared with those from nonsensitized control mice (p < 0. 05). After allergen challenge, there was a further increase in IL-5 expression (p < 0.05) within the bone marrow. Both sensitization and allergen challenge resulted in an increase in the number of cells expressing major basic protein (MBP) (p < 0.05). In nonsensitized mice, the IL-5 mRNA was expressed predominantly by CD34-positive (CD34+) progenitor cells. Following sensitization and allergen challenge, CD3-positive (CD3+) T lymphocytes were the major source of this cytokine. These results demonstrate the presence of IL-5 within the bone marrow of normal Balb/c mice. After sensitization and allergen challenge, the increase in IL-5-producing cells within the bone marrow is attributed by T lymphocytes. PMID- 9731031 TI - Hypoxic stimulation of the stress-activated protein kinases in pulmonary artery fibroblasts. AB - Pulmonary hypertension in response to chronic hypoxia is invariably accompanied by remodeling of the pulmonary vessels but the mechanism by which hypoxia increases the replication of vascular cells is unknown. To investigate the hypothesis that hypoxia stimulates intracellular kinase cascades we measured the activity of "classic" mitogen-activated protein (MAP) kinase pathways and "stress activated" MAP kinase pathways in bovine pulmonary artery fibroblasts subjected to hypoxia for up to 30 h. Hypoxia (1% O2) stimulated strongly the stress activated protein kinases, c-Jun NH2-terminal kinase (JNK) and p38 MAP kinase. Two peaks of p38 MAP kinase activity at 6 and 24 h were associated with an increase in the activity of mitogen-activated protein kinase-activated protein (MAPKAP) kinase-2, the immediate downstream target of p38 MAP kinase. Furthermore, the second phase of p38 MAP kinase activity could be reversed if cells were reoxygenated after 12 h. These data suggest that hypoxic stimulation of pulmonary artery cells is mediated by activation of the stress-activated protein kinases. PMID- 9731032 TI - Glucocorticoid receptors in bronchial epithelial cells in asthma. AB - The expression of the glucocorticoid receptor (GR) in untreated or in steroid dependent asthmatic patients is poorly understood. We therefore studied GR mRNA and protein levels in bronchial biopsies obtained from seven untreated asthmatic patients, seven control volunteers, and seven patients with chronic bronchitis. We also studied in bronchial epithelial cells obtained by brushing from 13 untreated asthmatics, 18 steroid-dependent asthmatics, 11 control volunteers, and 12 patients with chronic bronchitis, GR and heat shock protein 90 kD (hsp90) mRNA as well as the immunoreactivity of GR, intercellular adhesion molecule (ICAM-1), and granulocyte macrophage-colony-stimulating factor (GM-CSF). GR mRNA and protein level was similar in all subject groups in both biopsies and bronchial epithelial cells. Hsp90 mRNA level was also similar in all subject groups. ICAM-1 expression was significantly increased in bronchial epithelial cells from untreated asthmatics, but ICAM-1 was not expressed in those from steroid dependent asthmatic patients. GM-CSF expression was significantly increased in bronchial epithelial cells from untreated and steroid-dependent asthmatic patients. GR expression within the airways is unaltered by oral long-term steroid treatment in asthma, but the expression of some but not all specific markers for asthma is modified by oral steroid. PMID- 9731033 TI - Talc-induced expression of C-C and C-X-C chemokines and intercellular adhesion molecule-1 in mesothelial cells. AB - Treatment of symptomatic carcinomatous pleural effusions is primarily directed at local palliation with a wide variety of sclerosing agents, of which talc is considered to be the most successful. The mechanism whereby talc achieves this effect is unknown. The objective of this study was to investigate whether talc stimulates pleural mesothelial cells (PMC) to release C-X-C and/or C-C chemokines and express adhesion molecules that initiate and amplify the inflammatory process in the pleural space. When PMC were challenged with talc in vitro, interleukin-8 (IL-8) and monocyte chemotactic protein-1 (MCP-1) levels were increased (p < 0.001) both at the protein and the mRNA level as compared with unstimulated cultures. Talc-stimulated PMC culture supernatant showed chemotactic activity for neutrophils and monocytes. The chemotactic activity of PMC culture supernatant was blocked by 44.2% with IL-8-specific antibody and by 55.7% with MCP-1-specific antibody, demonstrating that PMC-derived chemokines are bioactive. Talc also enhanced intercellular adhesion molecule-1 (ICAM-1) expression in PMC. The data demonstrate that talc stimulates PMC to release chemokines and express adhesion molecules that may play a critical role in pleurodesis. PMID- 9731034 TI - Pulmonary infection with Mycobacterium avium-intracellulare leads to air trapping distal to the small airways. AB - To clarify the structure and function of the airways in Mycobacterium avium intracellulare (MAI) infection, we performed pulmonary function tests and high resolution computed tomography (HRCT) of the thorax in female patients 61 +/- 9 yr of age (n = 12) with pulmonary MAI infection without predisposing lung disease and compared their data with those of normal female volunteers 54 +/- 8 yr of age (n = 9). We calculated the E/I ratio, i.e., the average ratio of HRCT number at full expiration to that at full inspiration, as an index for the evaluation of air trapping distal to the small airways. Patients showed significant increases in residual volume and slope of phase III (DeltaN2) of the single-breath nitrogen test, and significant decreases in flow at 50 and 25% of FVC, suggesting hyperinflation and obstruction of the small airways. HRCT of patients revealed the small nodules and ectasis of bronchioles and small bronchi located mainly in segments (S) S2, S3, S4, and S5. The E/I ratio was significantly elevated in patients, and especially higher in the upper lung field than in the lower lung field, suggesting air trapping distal to the small airways. The difference of E/I ratio between the upper and lower field is probably related to the segmental distribution of CT abnormalities. These findings suggest that MAI infection can lead to air trapping distal to the small airways. PMID- 9731035 TI - Bronchial mucosal immunoreactivity of sensory neuropeptides in severe airway diseases. AB - Neuropeptides act on most of the components of the bronchial environment. They influence bronchomotor tone and bronchial vascular caliber and permeability. To investigate the nonadrenergic, noncholinergic system within the airways in asthma and chronic bronchitis, we performed endobronchial biopsies in 16 normal human volunteers, 49 patients with asthma of varying severity, including 16 patients treated with oral corticosteroids, and 13 patients with chronic bronchitis. Frozen sections of biopsies stained with specific antibodies against the neural marker PGP 9.5, vasoactive intestinal peptide (VIP), substance P (SP), calcitonin gene-related peptide (CGRP), and neuropeptide Y (NPY) were analyzed for the presence of nerves through indirect immunofluorescence. Nerves were present in most of the biopsies and were found within and below the epithelium and adjacent to smooth muscle, glands, and blood vessels. By comparison with those in normal subjects, the numbers of VIP-immunoreactive nerves were not significantly decreased in patients with asthma and chronic bronchitis, but NPY-immunoreactive nerves were significantly decreased in the smooth muscle of these latter two groups of patients (p < 0.005). There was no correlation between disease severity and the number of nerves found in the biopsies. This study does not confirm previous findings in autopsy material of some defects in sensory and VIP containing nerves in severe asthma. PMID- 9731036 TI - Elevated levels of expired breath hydrogen peroxide in bronchiectasis. AB - Airway inflammation is important in the development and progression of many lung diseases, including bronchiectasis. Activation of inflammatory cells such as neutrophils, eosinophils, and macrophages induces a respiratory burst resulting in the production of reactive oxygen species such as hydrogen peroxide (H2O2). We have measured exhaled H2O2 in patients with documented bronchiectasis and investigated whether the concentration of H2O2 is related to the disease severity, as defined by lung function. We also investigated whether the concentrations of expired H2O2 were different in bronchiectatic patients who received inhaled corticosteroids compared with steroid-naive patients. In 37 patients with bronchiectasis (mean age, 45 +/- 2.5 yr; FEV1, 59 +/- 3% pred), mean H2O2 concentration in exhaled breath condensate was significantly elevated as compared with the values in 25 age-matched (mean age, 42 +/- 2 yr) normal subjects (0.87 +/- 0.01 versus 0.26 +/- 0.04 microM, p < 0.001). There was a significant negative correlation between H2O2 and FEV1 (r = -0.76, p < 0.0001). Patients treated with inhaled corticosteroids had values of H2O2 similar to those of steroid-naive patients (0.8 +/- 0.1 versus 0.9 +/- 0.1, p > 0.05). We conclude that H2O2 is elevated in exhaled air condensate of patients with bronchiectasis and is correlated with disease severity. Measurement of H2O2 may be used as a simple noninvasive method to monitor airway inflammation and oxidative stress. PMID- 9731037 TI - Surfactant homeostasis in corticotropin-releasing hormone deficiency in adult mice. AB - Glucocorticoids are essential for lung maturation and pharmacologic doses of glucocorticoids increase surfactant in adult rats. Therefore, we asked if glucocorticoid deficiency in corticotropin-releasing hormone-deficient mice (CRH /-) with very low plasma corticosterone levels would alter surfactant pool sizes and precursor incorporation into saturated phosphatidylcholine (Sat PC). Alveolar and lung tissue Sat PC pool sizes were not different for CRH-/- mice and wild type mice. The incorporation of [3H]choline into Sat PC also was similar for the two strains of mice. Glucocorticoids are not a major regulator of surfactant homeostasis in the adult mouse. PMID- 9731039 TI - Chronobiology of asthma. PMID- 9731038 TI - Detection of Mycoplasma pneumoniae in the airways of adults with chronic asthma. AB - Infection with Mycoplasma pneumoniae has been shown to exacerbate asthma in humans. However, the role of M. pneumoniae in the pathogenesis of chronic asthma has not been defined. Eighteen asthmatics with chronic, stable asthma and 11 nonasthmatic control subjects underwent evaluation of the upper and lower airways and serologic analysis to determine the presence of M. pneumoniae, Chlamydia pneumoniae, and seven respiratory viruses through culture, enzyme-linked immunoassay (EIA) and polymerase chain reaction (PCR). M. pneumoniae was detected by PCR in 10 of 18 asthmatics and one of 11 control subjects (p = 0.02). In nine of the 10 patients, the organism was detected in bronchoalveolar lavage or bronchial biopsies. Seven of 18 asthmatics and one of 11 control subjects were also positive for M. fermentans and M. genitalium by PCR. All patients' cultures, EIAs, and serology were negative for M. pneumoniae. All PCR and cultures were negative for C. pneumoniae, and all EIAs for respiratory viruses were negative in all subjects. Nine asthmatics and one control subject exhibited positive serology for C. pneumoniae (p = 0.05). M. pneumoniae was present in the lower airways of chronic, stable asthmatics with greater frequency than control subjects, and may play a role in the pathogenesis of chronic asthma. PMID- 9731040 TI - Iron is hot: an update on the pathophysiology of hemochromatosis. PMID- 9731042 TI - Activation of p38 MAP kinase and JNK but not ERK is required for erythropoietin induced erythroid differentiation. AB - p38 MAP kinase (p38) and JNK have been described as playing a critical role in the response to a variety of environmental stresses and proinflammatory cytokines. It was recently reported that hematopoietic cytokines activate not only classical MAP kinases (ERK), but also p38 and JNK. However, the physiological function of these kinases in hematopoiesis remains obscure. We found that all MAP kinases examined, ERK1, ERK2, p38, JNK1, and JNK2, were rapidly and transiently activated by erythropoietin (Epo) stimulation in SKT6 cells, which can be induced to differentiate into hemoglobinized cells in response to Epo. Furthermore, p38-specific inhibitor SB203580 but not MEK specific inhibitor PD98059 significantly suppressed Epo-induced differentiation and antisense oligonucleotides of p38, JNK1, and JNK2, but neither ERK1 nor ERK2 clearly inhibited Epo-induced hemoglobinization. However, in Epo-dependent FD-EPO cells, inhibition of either ERKs, p38, or JNKs suppressed cell growth. Furthermore, forced expression of a gain-of-function MKK6 mutant, which specifically activated p38, induced hemoglobinization of SKT6 cells without Epo. These results indicate that activation of p38 and JNKs but not of ERKs is required for Epo-induced erythroid differentiation of SKT6 cells, whereas all of these kinases are involved in Epo-induced mitogenesis of FD-EPO cells. PMID- 9731041 TI - Collagen induces tyrosine phosphorylation of Wiskott-Aldrich syndrome protein in human platelets. AB - Wiskott-Aldrich syndrome (WAS) and X-linked thrombocytopenia (XLT) are caused by mutations of the WAS protein (WASP) gene. All hematopoietic stem cell-derived lineages, including platelets, express WASP. Platelets from WAS patients are smaller than their normal counterparts and defects in platelet aggregation and actin polymerization have been reported. To determine if WASP is important for normal platelet function, we examined its role in signal transduction. We found that collagen but not thrombopoietin or thrombin induces a rapid and robust increase in tyrosine phosphorylation of platelet-associated WASP. Collagen induced tyrosine phosphorylation of WASP was inhibited by cytochalasin D and wortmannin, respectively, suggesting that actin polymerization and phosphatidylinositol 3-kinase (PI3-kinase) play a role in the induction of tyrosine phosphorylation of WASP. Binding of glutathion S-transferase (GST)-Grb2 to WASP was seen in the lysate of resting platelets. The binding was reduced when lysates from collagen-stimulated platelets were incubated with GST-Grb2, suggesting that tyrosine phosphorylation of WASP may directly or indirectly modulate the adapter function of WASP. Although thrombin- and thrombopoietin induced increase in tyrosine phosphorylation of WASP is negligible or marginal, WASP from thrombin-activated platelets became incorporated into the Triton X-100 insoluble 10, 000g sedimentable residue in an aggregation-dependent manner, suggesting that it may have a regulatory role in platelet cytoskeletal processes during aggregation. Lastly, we found that WASP is cleaved in response to activation of calpain, a protease that may have a role in postaggregation signaling processes. Our data suggest that collagen specifically induces an increase in tyrosine phosphorylation of WASP and that WASP is involved in signaling during thrombin-induced aggregation by its redistribution to the cytoskeleton and its cleavage during aggregation. PMID- 9731043 TI - Increased susceptibility in Hp knockout mice during acute hemolysis. AB - Haptoglobin, a conserved plasma glycoprotein, forms very stable soluble complexes with free plasma hemoglobin. Hemoglobin binding by haptoglobin is thought to be important in the rapid hepatic clearance of hemoglobin from the plasma and in the inhibition of glomerular filtration of hemoglobin. To evaluate these functions, Haptoglobin knockout (-/-) mice were created. These mice were viable but had a small, significant reduction in postnatal viability. Contrary to popular belief, the lack of haptoglobin did not impair clearance of free plasma hemoglobin in -/- mice. Induction of severe hemolysis by phenylhydrazine caused extensive hemoglobin precipitation in the renal tubular cells of both -/- and +/+ mice, with death occurring in 55% of -/- mice and in 18% of +/+ mice. In general, phenylhydrazine-treated -/- mice suffered greater tissue damage, as evidenced by the induction of hepatic acute phase response resulting in increased plasma alpha 1-acid glycoprotein (AGP) levels. Among -/- and +/+ mice that survived, -/- mice tend to suffer greater oxidative damage and failed to repair or regenerate damaged renal tissues, as indicated by their higher plasma malonaldehyde (MDA) and 4-hydroxy-2(E)-nonenal (HNE) levels and lower mitotic indices in their kidneys, respectively. This study suggested that a physiologically important role of hemoglobin-haptoglobin complex formation is the amelioration of tissue damages by hemoglobin-driven lipid peroxidation. PMID- 9731044 TI - Improved gene transfer into baboon marrow repopulating cells using recombinant human fibronectin fragment CH-296 in combination with interleukin-6, stem cell factor, FLT-3 ligand, and megakaryocyte growth and development factor. AB - We have used a competitive repopulation assay in baboons to develop improved methods for hematopoietic stem cell transduction and have previously shown increased gene transfer into baboon marrow repopulating cells using a gibbon ape leukemia virus (GALV)-pseudotype retroviral vector (Kiem et al, Blood 90:4638, 1997). In this study using GALV-pseudotype vectors, we examined additional variables that have been reported to increase gene transfer into hematopoietic progenitor cells in culture for their ability to increase gene transfer into baboon hematopoietic repopulating cells. Baboon marrow was harvested after in vivo administration (priming) of stem cell factor (SCF) and granulocyte colony stimulating factor (G-CSF). CD34-enriched marrow cells were divided into two equal fractions to directly compare transduction efficiencies under different gene transfer conditions. Transduction by either incubation with retroviral vectors on CH-296-coated flasks or by cocultivation on vector-producing cells was studied in five animals; in one animal, transduction on CH-296 was compared with transduction on bovine serum albumin (BSA)-coated flasks. The highest level of gene transfer was obtained after 24 hours of prestimulation followed by 48 hours of incubation on CH-296 in vector-containing medium in the presence of multiple hematopoietic growth factors (interleukin-6, stem cell factor, FLT-3 ligand, and megakaryocyte growth and development factor). Using these conditions, up to 20% of peripheral blood and marrow cells contained vector sequences for more than 20 weeks, as determined by both polymerase chain reaction and Southern blot analysis. Gene transfer rates were higher for cells transduced on CH-296 as compared with BSA or cocultivation. In one animal, we have used a vector expressing a cell surface protein (human placental alkaline phosphatase) and have detected 10% and 5% of peripheral blood cells expressing the transduced gene 2 and 4 weeks after transplantation as measured by flow cytometry. In conclusion, the conditions described here have resulted in gene transfer rates that will allow detection of transduced cells by flow cytometry to facilitate the evaluation of gene expression. The levels of gene transfer obtained with these conditions suggest the potential for therapeutic efficacy in diseases affecting the hematopoietic system. PMID- 9731045 TI - Eotaxin modulates myelopoiesis and mast cell development from embryonic hematopoietic progenitors. AB - Eotaxin is a potent chemoattractant for eosinophils during inflammation and allergic reactions in the adult, but its role in the embryonic development of the hematopoietic system has not been examined. We report here that eotaxin and its receptor, CCR-3, are expressed by embryonic tissues responsible for blood development, such as fetal liver (FL), yolk sac (YS), and peripheral blood. We found that eotaxin acts synergistically with stem cell factor to accelerate the differentiation of embryonic mast cell progenitors, and this response can be suppressed by pertussis toxin, an inhibitor of chemokine-induced signaling through Gialpha protein and chemotaxis. Eotaxin promotes the differentiation of fetal mast cell progenitors into differentiated mast cells as defined by the expression of mast cell specific proteases. Furthermore, in combination with stem cell factor (SCF), it promotes the growth of Mac-1(+) myeloid cells from embryonic progenitors. These studies suggest that eotaxin may be involved in the growth of granulocytic progenitors and the differentiation and/or function of mast cells during embryogenesis and/or pathological conditions that induce high levels of eotaxin, such as allergic responses. PMID- 9731046 TI - Immunophenotypic and genotypic features, clinical characteristics, and treatment outcome of adult pro-B acute lymphoblastic leukemia: results of the German multicenter trials GMALL 03/87 and 04/89. AB - In contrast to childhood acute lymphoblastic leukemia (ALL), the cell-biological features, clinical characteristics, and treatment outcome of CD10(-) pro-B ALL have not yet been determined in larger series of adult patients. Therefore, we studied 57 adult patients with newly diagnosed pro-B ALL (median age, 30 years) enrolled in two consecutive German multicenter ALL studies (03/87 and 04/89). Extensive immunophenotypic characterization of leukemic blasts could be performed on all patients, whereas adequate cytogenetic data were available in 33 cases and molecular studies in 18 cases, using reverse transcription-polymerase chain reaction to detect MLL-AF-4 transcripts. Twenty-two patients demonstrated a t(4;11)(q21;q23) and/or MLL-AF-4 rearrangements, and 6 patients had other structural abnormalities, including a t(9;22)(q34;q11) (N = 2). Nine patients had a normal karyotype. Patients with 11q23 abnormalities tended to be younger (median age, 29 years) and were characterized by male predominance (64%), hyperleukocytosis (median leukocyte count, 168 x 10(9)/L), and a frequent coexpression of CD65s (64%) as compared with patients with other cytogenetic abnormalities or a normal karyotype. Twelve of 16 (75%) pro-B ALL patients in study 03/87 and 30 of 41 (73%) in study 04/89 achieved a complete remission (CR). Sixteen of 30 patients in study 04/89 remain in continuous CR (CCR) in contrast to only 2 of 12 patients in study 03/87. Interestingly, all 7 patients treated with high-dose cytarabine and mitoxantrone as consolidation in study 04/89 remain alive and leukemia-free. One patient in study 03/87 and 8 in study 04/89 underwent autologous (N = 2) or allogeneic (N = 7) bone marrow transplantation (BMT). The median remission duration was 420 days for patients in study 03/87 and has not yet been reached in study 04/89. The median survival time of all pro-B ALL patients was 571 days in study 03/87 and 747 days in study 04/89. Among the 22 patients with a t(4;11) and/or MLL-AF-4 rearrangements, 17 achieved a CR and 8 are still in CCR, of whom 4 underwent an allogeneic BMT. Remission duration and overall survival did not differ significantly between pro-B ALL patients with 11q23 abnormalities and those with a normal karyotype or other structural abnormalities. These data indicate that intensification of postremission treatment may improve the prognosis of adult pro-B ALL, including patients with a t(4;11). PMID- 9731047 TI - Cytogenetic abnormalities in primary myelodysplastic syndrome are highly predictive of outcome after allogeneic bone marrow transplantation. AB - Allogeneic bone marrow transplantation (BMT) is the only curative therapy available for patients with myelodysplastic syndrome (MDS). In an attempt to identify prognostic factors influencing outcome, we collected data retrospectively on 60 consecutive adult patients who had undergone BMT at our center for primary MDS or acute myelogenous leukemia evolving from preexisting primary MDS (sAML). Patients were divided into subgroups according to cytogenetic abnormalities based on a recently described International MDS Workshop categorization system. The 7-year actuarial event-free survival (EFS), relapse rate, and nonrelapse mortality (NRM) for all patients were 29% (95% confidence interval [CI], 16% to 43%), 42% (CI, 24% to 67%), and 50% (CI, 37% to 64%), respectively. The EFS for the good-, intermediate-, and poor-risk cytogenetic subgroups were 51% (CI, 30% to 69%), 40% (CI, 16% to 63%), and 6% (CI, 0% to 24%), respectively (P = .003). The corresponding actuarial relapse rates were 19% (CI, 6% to 49%), 12% (CI, 2% to 61%), and 82% (CI, 48% to 99%), respectively (P = . 002) with no difference in NRM between the subgroups. Univariate analysis showed cytogenetic category, French-American-British (FAB) subtype, and graft versus-host disease (GVHD) prophylaxis used to be predictive of relapse and EFS. In multivariate analysis, only the cytogenetic category was predictive of EFS, with the relative risk of treatment failure for the good-, intermediate-, and poor-risk cytogenetic subgroups being 1.0, 1.5, and 3.5, respectively (P = . 004). For adults with primary MDS and sAML, even after BMT, poor-risk cytogenetics are predictive of an unfavorable outcome; novel treatment strategies will be required to improve results with allogeneic BMT in this patient population. PMID- 9731048 TI - Long-term follow-up of patients with hairy cell leukemia after cladribine treatment. AB - Hairy cell leukemia is a chronic B-cell disorder that follows an indolent, but progressive course. Cladribine (2-chlorodeoxyadenosine) induces complete remissions in the majority of patients after a single course. We report the long term outcomes, including response rates and their duration; time-to-treatment failure (TTF) rates; retreatment results; toxicities; and survival rates of patients treated at Scripps Clinic (La Jolla, CA). A total of 358 patients with hairy cell leukemia were treated with cladribine at 0.087 or 0.1 mg/kg body weight per day by continuous intravenous infusion for 7 days. The expected number of second neoplasms was based on the National Cancer Institute's Surveillance Epidemiology and End Results data. Of 349 evaluable patients, 319 (91%) achieved an initial complete response and 22 (7%) a partial response with an overall median duration of response follow-up of 52 months. Ninety patients (26%) had relapsed at a median of 29 months. The TTF rate for all 341 responders was 19% at 48 months, 16% for complete responders, and 54% for partial responders. Of 53 evaluable patients treated with second courses of cladribine at first relapse, 33 (62%) achieved complete responses and 14 (26%) partial responses. Twenty-seven patients (8%) developed second neoplasms (only 1 hematopoietic) with an observed to-expected ratio of 1.88 (95% confidence interval, 1.24 to 2.74). The overall survival rate was 96% at 48 months. Single courses of cladribine induced long lasting complete responses in the vast majority of patients. Relapse rates for complete responders were low. Patients who relapse can be successfully retreated with cladribine. Cladribine has high efficacy and a favorable acute and long-term toxicity profile when administered to patients with hairy cell leukemia. PMID- 9731049 TI - Rituximab (anti-CD20 monoclonal antibody) for the treatment of patients with relapsing or refractory aggressive lymphoma: a multicenter phase II study. AB - Rituximab, a chimeric monoclonal antibody that binds specifically to the CD20 antigen, induced objective responses in 50% of patients with low-grade or follicular B-cell lymphoma. Because most nonfollicular B-cell lymphomas also express the CD20 antigen, we conducted a phase II study to evaluate the efficacy and tolerability of this new agent in patients with more aggressive types of lymphoma. Patients with diffuse large B-cell lymphoma (DLCL), mantle cell lymphoma (MCL), or other intermediate- or high-grade B-cell lymphomas according to the Working Formulation were included in this prospective randomized phase II study if they were in first or second relapse, if they were refractory to initial therapy, if they progressed after a partial response to initial therapy, or if they were elderly (age >60 years) and not previously treated. The patients received 8 weekly infusions of rituximab at the dose of 375 mg/m2 in arm A or one infusion of 375 mg/m2 followed by 7 weekly infusions of 500 mg/m2 in arm B. Patients were evaluated 2 months after the last rituximab infusion. Fifty-four patients were randomized from 9 centers in Europe and Australia (28 in arm A and 26 in arm B). A total of 5 complete responses (CR) and 12 partial responses (PR) were observed among the 54 enrolled patients, with no difference between the two doses. In an intent-to-treat analysis, the CR rate was 9% (CI95%, 3% to 20%) and the PR rate was 22% (CI95%, 12% to 36%), for an overall response rate of 31% (CI95%, 20% to 46%). An analysis of prognostic factors showed that response rates were lower in patients with refractory disease, patients with lymphoma not classified as DLCL, and patients with a tumor larger than 5 cm in diameter. DLCL and MCL patients had response rates of 37% and 33%, respectively. The median time to progression exceeded 246 days for the 17 responding patients. The most frequently reported adverse events were related to an infusion syndrome and were mild: 19% of the patients had a grade 3 related adverse event, slightly more in arm B, and only 1 patient had a grade 4 related adverse event in arm A. Two patients (3.7%) withdrew from treatment because of severe adverse events, one patient in each arm. In this first trial of rituximab in DLCL and MCL, patients experienced a significant clinical activity with a low toxicity. Rituximab has significant activity in DLCL and MCL patients and should be tested in combination with chemotherapy in such patients. PMID- 9731050 TI - Treatment-related deaths and second cancer risk after autologous stem-cell transplantation for Hodgkin's disease. AB - Autologous stem-cell transplantation has become a widely used therapy in Hodgkin's disease (HD). To appreciate the early and late risks associated with this procedure, its lethal toxicity and effects on the incidence of secondary cancers were studied. Data related to 467 French patients grafted from 1982 to 1995 for primary sensitive disease (PSD, 22%), primary refractory disease (PRD, 18%), first relapse (R1, 45%), or subsequent relapses (R2, 15%) were analyzed. Grafted patients (PSD, PRD, and R1; n = 393) were matched (3 controls for 1 case) on age, gender, clinical stage, B symptoms, and time at risk with 1179 conventionally treated patients issued from international databases. The proportional hazards (Cox) model was used to assess relative risks (RR). Among grafted patients, 8% died of toxicity related to the procedure, and 18 secondary cancers occurred leading to a 5-year cumulative incidence rate of 8.9%. In this series, risk factors for second cancer were age >/=40 years (RR = 3.73, P = .007) and the use of peripheral blood stem cells as source of graft (RR = 3.10, P = .03). Among grafted and matched ungrafted patients, risk factors for the development of secondary cancer were age >/=40 years (RR = 2.90, P < .001), relapse versus no relapse (RR = 5.22, P = .006), PRD versus other patients (RR = 3.86, P = .033), and grafted versus ungrafted patients (RR = 2.04, P = . 024). Solid tumors were more frequent in grafted than in ungrafted patients (RR = 5.19, P = .001) although the incidence of myelodysplasia and acute myeloid leukemia was similar in the two groups. We conclude that high-dose chemotherapy administered as first-line treatment or after relapse is associated with an acceptable toxic death rate. The risk of secondary myelodysplasia or acute myeloid leukemia is not significantly increased after autologous stem-cell transplantation for HD, whereas an increased risk of solid tumors exists. The peripheral blood stem-cell associated risk of secondary cancer among grafted patients needs further investigations. PMID- 9731051 TI - IL-2 adenovector-transduced autologous tumor cells induce antitumor immune responses in patients with neuroblastoma. AB - In many different murine models, the immunogenicity of tumor cells can be increased by transduction with a range of immunostimulatory genes, inducing an immune response that causes regression of pre-existing unmodified tumor cells. To investigate the relevance of these animal models to pediatric malignancy, we used autologous unirradiated tumor cells transduced with an adenovirus-IL-2 to immunize 10 children with advanced neuroblastoma. In a dose-escalation study, we found that this tumor immunogen induced a moderate local inflammatory response consisting predominantly of CD4(+) T lymphocytes, and a systemic response, with a rise in circulating CD25(+) and DR+ CD3(+) T cells. Patients also made a specific antitumor response, manifest by an IgG antitumor antibody and increased cytotoxic T-cell killing of autologous tumor cells. Clinically, five patients had tumor responses after the tumor immunogen alone (one complete tumor response, one partial response, and three with stable disease). Four of these five patients were shown to have coexisting antitumor cytotoxic activity, as opposed to only one of the patients with nonresponsive disease. These results show a promising correlation between preclinical observations and clinical outcome in this disease, and support further exploration of the approach for malignant diseases of children. PMID- 9731052 TI - Granulocyte colony-stimulating factor enhances bone marrow stem cell damage caused by repeated administration of cytotoxic agents. AB - Despite the increasing use of cytokines to circumvent the acute dose-limiting myelotoxicity of cancer treatment, little is known about the combined effects of cytotoxic agents and cytokines on the primitive stem cells responsible for long term hematopoiesis. In an experimental model, we administered cytotoxic agents that have variable effects on primitive stem cells in C57BL/6 (B6)-mice. Mice received six every-other-week doses of cyclophosphamide (CY, 84 mg/kg), VP-16 (24 mg/kg) + cisplatinum (2.4 mg/kg), carboplatinum (50 mg/kg), chlorambucil (12 mg/kg), BCNU (13.2 mg/kg), or TBI (80 cGy). Granulocyte colony-stimulating factor (G-CSF; 250 microg/kg/day) was administered subcutaneously twice daily on days 3 to 6 after each dose of the cytotoxic agent. Comparison with animals receiving the cytotoxic agent alone was made to investigate the effects of G-CSF on long term hematopoiesis. Hematopoiesis was measured 20 weeks after the last dose of the cytotoxic agent by assessment of peripheral blood counts, marrow cellularity, progenitor cell content (colony-forming units-spleen; CFU-S), and primitive stem cell number (long-term repopulating ability and day 28 and day 35 cobblestone area-forming cell [CAFC] frequencies). Exposure to cytotoxic agents alone resulted in a significant decrease in primitive stem cells (as measured by repopulating units [RU] and day 28 and day 35 CAFC content) in animals given carboplatinum, chlorambucil, BCNU, and TBI, but not in animals treated with cyclophosphamide or VP-16 and cisplatinum. The addition of G-CSF resulted in a significant decrease in stem cell content when compared with no G-CSF administration in animals treated with chlorambucil, BCNU, or TBI. Thus, G-CSF administered after repeated exposure to cytotoxic agents, appeared to damage the primitive stem cell compartment when used in combination with agents known to damage primitive stem cells. These results, although obtained in an experimental model, should raise concerns for the indiscriminate use of G-CSF in the clinic. PMID- 9731053 TI - The zinc finger transcription factor Egr-1 activates macrophage differentiation in M1 myeloblastic leukemia cells. AB - We previously have shown that the zinc finger transcription factor Egr-1 blocked granulocytic differentiation of HL-60 cells, restricting differentiation along the monocytic lineage. Egr-1 also was observed to block granulocyte colony stimulating factor (G-CSF)-induced differentiation of interleukin-3 (IL-3) dependent 32Dcl3 hematopoietic precursor cells, endowing the cells with the ability to be induced by granulocyte-macrophage colony-stimulating factor (GM CSF) for terminal differentiation along the macrophage lineage. To better understand the function of Egr-1 as a positive modulator of monocytic differentiation, in this work we have studied the effect of ectopic expression of Egr-1 on the murine myeloblastic leukemic cell line M1, which is induced for differentiation by the physiological inducer IL-6. It is shown that, unlike in HL 60 and 32Dcl3 cells, ectopic expression of Egr-1 in M1 cells resulted in activation of the macrophage differentiation program in the absence of differentiation inducer. This included the appearance of morphologically differentiated cells, decreased growth rate in mass culture, and cloning efficiency in soft agar, and expression of endogenous c-myb and c-myc mRNAs was markedly downregulated. Untreated M1Egr-1 cells also exhibited cell adherence, expression of Fc and C3 receptors, and upregulation of the myeloid differentiation primary response genes c-Jun, junD, and junB and the late genetic markers ferritin light-chain and lysozyme. Ectopic expression of Egr-1 in M1 cells also dramatically increased the sensitivity of the cells for IL-6-induced differentiation, allowed a higher proportion of M1 cells to become terminally differentiated under conditions of optimal stimulation for differentiation, and decreased M1 leukemogenicity in vivo. These findings demonstrate that the functions of Egr-1 as a positive modulator of macrophage differentiation vary, depending on the state of lineage commitment for differentiation of the hematopoietic cell type. PMID- 9731054 TI - Interleukin-10 inhibits erythropoietin-independent growth of erythroid bursts in patients with polycythemia vera. AB - In polycythemia vera (PV) erythroid colonies that grow in vitro in the absence of exogenous erythropoietin (EPO) arise from the abnormal clone that is responsible for overproduction of red blood cells. Although the mechanism of autonomous formation of burst-forming units-erythroid (BFU-E) is not fully understood, a spontaneous release of growth regulatory molecules by PV cells and/or by accessory cells is likely to be involved. Because of its cytokine synthesis inhibiting action, interleukin-10 (IL-10) could be a potentially useful molecule to modulate abnormal erythropoiesis in PV. We studied the effect of recombinant human IL-10 on the EPO-independent growth of erythroid bursts derived from peripheral blood mononuclear cells (PBMNCs) of patients with PV. IL-10 showed a profound, dose-dependent, and specific inhibitory effect on autonomous BFU-E formation. Ten nanograms per milliliter of IL-10 significantly suppressed spontaneous growth of erythroid colonies in methylcellulose in five of five PV patients tested with a mean inhibition by 81% (range, 72-94). To elucidate the possible mechanism of the inhibitory action of IL-10 we further studied the effect of anticytokine antibodies on autonomous BFU-E growth and the ability of exogenous cytokines to restore IL-10-induced suppression of erythroid colony growth. Among a panel of growth regulatory factors tested (granulocyte-macrophage colony-stimulating factor [GM-CSF], IL-3, granulocyte colony-stimulating factor, stem cell factor, and insulin-like growth factor-1) GM-CSF was the only molecule for which both an inhibition of spontaneous BFU-E formation by its respective antibody as well as a significant restimulation of erythroid colonies in IL-10 treated cultures by exogenous addition was found. Moreover, inhibition of GM-CSF production by IL-10 was shown in PV PBMNCs at the mRNA level. Our data indicate that autonomous BFU-E growth in PV can be profoundly inhibited by IL-10 and that this inhibitory effect seems to be at least in part secondary to suppression of endogenous GM-CSF production. PMID- 9731055 TI - Impaired expression of integrin alpha-4 subunit in cultured mast cells derived from mutant mice of mi/mi genotype. AB - The mi locus encodes a member of the basic-helix-loop-helix-leucine zipper protein family of transcription factors (hereafter called MITF). We have reported that expression of several genes was impaired in cultured mast cells (CMCs) of mi/mi mice due to a defective transactivation ability of mutant MITF (mi-MITF). Because attachment of mi/mi CMCs to fibroblasts is impaired, we examined the expression of integrin genes in mi/mi CMCs in the present study. Among the integrin genes examined, the expression of integrin alpha4 subunit was barely detectable in mi/mi CMCs, and the alpha4 protein was not detected by flow cytometry either. The specific adhesion to vascular cell adhesion molecule-1 (VCAM-1), the ligand for alpha4 subunit, was observed in +/+ CMCs but not in mi/mi CMCs, indicating that the expression of integrin alpha4 subunit at a functional level did not occur in mi/mi CMCs. In the promoter region of the alpha4 subunit gene, there was a CACTTG motif to which normal MITF (+- MITF) bound. The coexpression of +-MITF but not of mi-MITF transactivated the promoter of the alpha4 subunit gene. The deletion or mutation of the CACTTG motif abolished the transactivation by +-MITF, suggesting that +-MITF directly transactivated the gene encoding alpha4 subunit of integrin. PMID- 9731056 TI - An agonist murine monoclonal antibody to the human c-Mpl receptor stimulates megakaryocytopoiesis. AB - Thrombopoietin (TPO) is a hematopoietic growth factor that stimulates megakaryocytopoiesis and platelet production in vivo and promotes the development of identifiable megakaryocytes in vitro. We have developed a murine monoclonal antibody, BAH-1, raised against human megakaryocytic cells, which specifically recognizes the c-Mpl receptor and shows agonist activity by stimulating megakaryocytopoiesis in vitro. BAH-1 antibody specifically binds to platelets and to recombinant c-Mpl with high affinity. Similar to TPO, BAH-1 alone supported the formation of colony-forming unit-megakaryocyte (CFU-MK) colonies. The combination of BAH-1 plus interleukin-3 or of BAH-1 plus human TPO significantly increased the number of human CFU-MK colonies. In addition, BAH-1 monoclonal antibody stimulated the proliferation and maturation of primary bone marrow megakaryocytes in a dynamic heterogeneous liquid culture system. Individual large megakaryocytes as well as small megakaryocytic cells were observed in cultures of CD34(+) CD41(+) cells in the presence of BAH-1 antibodies. Similar to TPO, BAH-1 antibody induced a significant response of murine immature megakaryocytes as observed by an increase in the detectable numbers of acetylcholinesterase positive megakaryocytes. No effects of BAH-1 antibody were observed on colony forming unit-granulocyte-macrophage, burst-forming unit-erythroid, or colony forming unit-erythroid colonies. In vivo studies showed that BAH-1, alone or in combination with TPO, expands the numbers of megakaryocytic progenitor cells in myelosuppressed mice. This antibody should prove useful in understanding the structure-function aspects of the c-Mpl receptor as well as in evaluating the effects of the sustained activation of this receptor in preclinical models of severe thrombocytopenia. PMID- 9731057 TI - Saturation mutagenesis of the beta subunit of the human granulocyte-macrophage colony-stimulating factor receptor shows clustering of constitutive mutations, activation of ERK MAP kinase and STAT pathways, and differential beta subunit tyrosine phosphorylation. AB - The high-affinity receptors for human granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-3 (IL-3), and IL-5 are heterodimeric complexes consisting of cytokine-specific alpha subunits and a common signal-transducing beta subunit (hbetac). We have previously demonstrated the oncogenic potential of this group of receptors by identifying constitutively activating point mutations in the extracellular and transmembrane domains of hbetac. We report here a comprehensive screen of the entire hbetac molecule that has led to the identification of additional constitutive point mutations by virtue of their ability to confer factor independence on murine FDC-P1 cells. These mutations were clustered exclusively in a central region of hbetac that encompasses the extracellular membrane-proximal domain, transmembrane domain, and membrane proximal region of the cytoplasmic domain. Interestingly, most hbetac mutants exhibited cell type-specific constitutive activity, with only two transmembrane domain mutants able to confer factor independence on both murine FDC-P1 and BAF B03 cells. Examination of the biochemical properties of these mutants in FDC-P1 cells indicated that MAP kinase (ERK1/2), STAT, and JAK2 signaling molecules were constitutively activated. In contrast, only some of the mutant beta subunits were constitutively tyrosine phosphorylated. Taken together, these results highlight key regions involved in hbetac activation, dissociate hbetac tyrosine phosphorylation from MAP kinase and STAT activation, and suggest the involvement of distinct mechanisms by which proliferative signals can be generated by hbetac. PMID- 9731058 TI - Leukemic predisposition of mice transplanted with gene-modified hematopoietic precursors expressing flt3 ligand. AB - flt3/flk-2 ligand (FL) is a cytokine that exhibits synergistic activities in combination with other early acting factors on subpopulations of hematopoietic stem/progenitor cells. In addition to normal hematopoietic precursors, expression of the FL receptor, flt3R, has been frequently demonstrated on the blast cells from patients with acute B-lineage lymphoblastic, myeloid, and biphenotypic (also known as hybrid or mixed) leukemias. Because many of these leukemic cell types express FL, the possibility has been raised that altered regulation of FL mediated signaling might contribute to malignant transformation or expansion of the leukemic clone. In humans, FL is predominantly synthesized as a transmembrane protein that must undergo proteolytic cleavage to generate a soluble form. To investigate the consequences of constitutively expressing the analogous murine FL isoform in murine hematopoietic stem/progenitor cells, lethally irradiated syngeneic mice (18 total) were engrafted with post-5-fluorouracil-treated bone marrow cells transduced ex vivo with a recombinant retroviral vector (MSCV-FL) encoding murine transmembrane FL. Compared with control mice (8 total), MSCV-FL mice presented with a mild macrocytic anemia but were otherwise healthy for more than 5 months posttransplant (until 22 weeks). Subsequently, all primary MSCV-FL recipients observed for up to 1 year plus 83% (20 of 24) of secondary MSCV-FL animals that had received bone marrow from asymptomatic primary hosts reconstituted for 4 to 5 months developed transplantable hematologic malignancies (with mean latency periods of 30 and 23 weeks, respectively). Phenotypic and molecular analyses indicated that the tumor cells expressed flt3R and displayed B cell and/or myeloid markers. These data, establishing that dysregulated expression of FL in primitive hematopoietic cells predisposes flt3R+ precursors to leukemic transformation, underscore a potential role of this cytokine/receptor combination in certain human leukemias. PMID- 9731059 TI - Ultrastructural analysis of bone marrow hematopoiesis in mice transgenic for the thymidine kinase gene driven by the alpha IIb promoter. AB - Transgenic mice have been generated with expression of the herpes virus thymidine kinase gene directed by a 2.7-kb fragment of the alphaIIb murine promoter of the gene encoding the alphaIIb-subunit of the platelet integrin alphaIIbbeta3 (Tropel et al, Blood 90:2995, 1997). Administration of ganciclovir (GCV) to these mice resulted not only in an acute cessation of platelet production due to the depletion of the megakaryocytic lineage, but also a decrease in erythrocyte and leukocyte numbers. Immunogold staining on ultrathin frozen sections and electron microscopy has now shown that the remaining population of immature hematopoietic cells contain a high proportion of Sca-1(+) and CD34(+) cells, with CD45R+ cells of the lymphopoietic lineage being maintained. Stromal cells were also preserved. Blood thrombopoietin levels were high. At 4 days of the recovery phase, Sca-1 and CD34 antigen expression decreased with intense proliferation of cells of the three lineages, with megakaryocyte (MK) progenitors being identified by their positivity for glycoprotein IIb-IIIa. These results suggest that transcriptional activity for the alphaIIb gene promoter was present on pluripotent hematopoietic stem cells. At 6 to 8 days after cessation of GCV, numerous mature MK were observed, some of them with deformed shapes crossing the endothelial barrier through thin apertures. Proplatelet production was visualized in the vascular sinus. After 15 days, circulating platelet levels had increased to approximately 65% of normal. Transgenic alphaIIb-tk mice constitute a valuable model to study in vivo megakaryocytopoiesis. PMID- 9731060 TI - Treatment of non-obese diabetic (NOD)/Severe-combined immunodeficient mice (SCID) with flt3 ligand and interleukin-7 impairs the B-lineage commitment of repopulating cells after transplantation of human hematopoietic cells. AB - Until recently, the identification of cellular factors that govern the developmental program of human stem cells has been difficult due to the absence of repopulation assays that detect human stem cells. The transplantation of human bone marrow (BM) or cord blood (CB) into non-obese diabetic (NOD)/severe-combined immunodeficient (SCID) mice has enabled identification of primitive human cells capable of multilineage repopulation of NOD/SCID mice (termed the SCID repopulating cell [SRC]). Here, we examined the effect of long-term in vivo treatment with various combinations of human cytokines on the developmental program of SRC. Detailed flow cytometric analysis of engrafted mice indicated that the vast majority of the human graft of untreated mice was comprised of B lymphocytes at various stages of development as well as myeloid and primitive cells; T cells were not reproducibly detected. Many studies, including murine in vitro and in vivo data and human in vitro experiments, have suggested that flt3 ligand (FL) and/or Interleukin-7 (IL-7) promotes T- and B-cell development. Unexpectedly, we found that treatment of engrafted mice with the FL/IL-7 combination did not induce human T- or B-cell development, but instead markedly reduced B-cell development with a concomitant shift in the lineage distribution towards the myeloid lineage. Effects on lineage distribution were similar in engrafted mice transplanted with highly purified cells indicating that the action of the cytokines was not via cotransplanted mature cells from CB or BM cells. These data show that the lineage development of the human graft in NOD/SCID mice can be modulated by administration of human cytokines providing a valuable tool to evaluate the in vivo action of human cytokines on human repopulating cells. PMID- 9731061 TI - Stimulation of mouse and human primitive hematopoiesis by murine embryonic aorta gonad-mesonephros-derived stromal cell lines. AB - We report here on a novel stromal cell line, AGM-S3, derived from the aorta-gonad mesonephros (AGM) region of a 10.5 days postcoitum (dpc) mouse embryo. The AGM-S3 cells promoted production of hematopoietic progenitors and day-12 spleen colony forming cells from Lin-c-Kit+Sca-1(+) murine primitive hematopoietic cells. They also supported for 6 weeks generation of human multipotential progenitors from cord blood CD34(+)CD38(-) primitive hematopoietic cells. Human long-term repopulating hematopoietic stem cells (LTR-HSC) with the potential to reconstitute hematopoiesis in NOD/SCID mice were maintained on AGM-S3 cells for at least 4 weeks. Flow cytometric analysis showed that CD13, vascular cellular adhesion molecule-1, and Sca-1 were expressed on AGM-S3 cells. Because stem cell factor, interleukin-6 (IL-6), and oncostatin M, but not IL-3, IL-11, leukemia- inhibitory factor, granulocyte colony-stimulating factor, granulocyte-macrophage colony-stimulating factor, thrombopoietin, and Flk2 ligand were detected in reverse transcription-polymerase chain reaction analysis of AGM-S3 cells, the cells seem to express species-cross reactive molecule(s) other than the cytokines examined and which act on primitive hematopoietic progenitor/stem cells. This cell line is expected to elucidate molecular mechanisms regulating early hematopoiesis and pave the way for developing strategies for expansion of human transplantable HSC. PMID- 9731062 TI - Apoptotic regulation in primitive hematopoietic precursors. AB - Bcl-2 and bcl-xL function as suppressors of programmed cell death. The expression of bcl-2 protein in vivo is associated with long-lived hematopoietic cells such as mature lymphocytes and early myeloid progenitors. Bcl-xL, a homologue of bcl 2, is also expressed in lymphocytes and thymocytes. In contrast, the bcl-2 related proteins (bax, bad, and bak) act by promoting apoptotic cell death as shown from their expression in hematopoietic cell lines. We analyzed the expression of bcl-2 and bcl-x proteins in hematopoietic precursors obtained from various cell sources in adult mobilized peripheral blood collected from 13 patients with solid tumors, 8 adult bone marrow, and 12 umbilical cord blood. The analysis was based on the expression of the proliferation and activation specific antigens, CD38 and class II (HLA-DR). Similarly, we analyzed the expression of bcl-2-related proteins bcl-xL, bax, bad, and bak before and during ex-vivo expansion. Hematopoietic precursors expressing strongly the CD34 antigen (CD34(s+)) and lacking CD38 or HLA-DR expression were analyzed by using three color immunofluorescence staining. The majority of CD34(+) cells expressed bcl-2 and unexpectedly showed a bimodal distribution of low and high expression. More cells that lacked or expressed low density CD38 expressed low bcl-2 than the more differentiated counterparts (those with high density CD38). Immaturity (ie, little or no HLA-DR) is associated with the expression of low bcl-2 compared with HLA-DR+. However, HLA-DR-/low population contained a lower number of cells expressing low bcl-2 (30% to 40%) than CD38(-/low) in comparable samples. The hematopoietic precursors with bcl-2(low) and bcl-2(high) formed a homogeneous population of undifferentiated lymphoid-like cells having a similar forward scatter. These cells expressed strongly the bcl-xL protein (>95%) but were bax low (4% to 12%), bad low (0% to 0.8%), and bak low (0% to 3%). The expression of apoptosis specific protein (ASP) was also low (3.4% +/- 3.1%) as was Annexin V. In addition, the CD34(+)/CD38(-) showed low cell cycle activity (<2.2%). Induction of apoptosis by overnight incubation of CD34 cells in serum-deprived medium resulted in the upregulation of bcl-2 as a single population histogram. Thus, these results suggest that in quiescent hematopoietic precursors, the bcl-2 protein plays a less prominent role as a survival promoter than bcl-xL and that the low bcl-2 expression did not promote apoptosis. During day 10 of ex vivo expansion of CD34(+) cells in liquid culture containing stem cell factor, interleukin-3 (IL-3), IL-6, IL-1beta, and erythropoietin, the CD34(+)/CD38(-) cells expressed high bcl-2 as a single population histogram, and greater than 90% were bcl-xL high. However, the expression of pro- and apoptotic antigens increased: bax (10% to 15%), bad (5% to 8%), bak (6% to 14%), and ASP (6% to 10%). These results show the importance of monitoring the expression of these proteins when defining the culture conditions for ex vivo expansion. PMID- 9731063 TI - A recombinant soluble form of the integrin alpha IIb beta 3 (GPIIb-IIIa) assumes an active, ligand-binding conformation and is recognized by GPIIb-IIIa-specific monoclonal, allo-, auto-, and drug-dependent platelet antibodies. AB - The IIb-IIIa glycoprotein complex is a favored target for allo-, auto-, and drug dependent antibodies associated with immune thrombocytopenia. A soluble, recombinant form of the GPIIb-IIIa heterodimer that could be produced in large quantities and maintained in solution without detergent could provide a useful experimental tool for the study of platelet-reactive antibodies, but previous attempts to produce such a construct have yielded only small quantities of the end product. Using a baculovirus expression system and the dual-promoter transfer vector P2Bac, we were able to express soluble GPIIb-IIIa complex (srGPIIb-IIIa) lacking cytoplasmic and transmembrane domains in quantities of about 1,000 microg/L, about 40 times greater than reported previously. The high yield achieved may be related to inclusion of the entire extracellular region of the GPIIb light chain in the construct. srGPIIb-IIIa reacts spontaneously with fibrinogen, and this interaction is totally inhibited by the peptide RGDS. Reactions of 24 GPIIb-IIIa-specific antibodies evaluated (12 monoclonal, 3 allo specific, 3 auto-specific, and 6 drug-dependent) with srGPIIb-IIIa were indistinguishable from reactions with platelet GPIIb-IIIa. Thus, srGPIIb-IIIa spontaneously assumes an active, ligand-binding conformation and contains epitopes for all monoclonal and human antibodies tested to date. srGPIIb-IIIa can be produced in large quantities, can readily be modified by site-directed mutagenesis, and should facilitate identification of epitopes recognized by GPIIb IIIa-specific antibodies, study of the mechanism(s) by which certain drugs promote antibody binding to GPIIb-IIIa in drug-induced thrombocytopenia and structure-function relationships of GPIIb-IIIa. PMID- 9731064 TI - No effect of clot age or thrombolysis on argatroban's inhibition of thrombin. AB - The purpose of this study was to establish the effects of clot age and thrombolysis, with either streptokinase or tissue-type plasminogen activator (tPA), on argatroban's ability to inhibit thrombin. The antithrombotic activity of argatroban has been quantified in fibrin clot permeation and fibrin clot perfusion systems as a function of clot age and composition. Analysis of the argatroban dose-response data with a competitive inhibition model has yielded IC50 values in the low micromolar range. Results obtained in a plasma clot permeation system have also shown that argatroban is a potent inhibitor of clot bound thrombin, independent of either clot age or the presence of hemostatically active platelets. Treatment of aged plasma clots with either streptokinase or alteplase, at therapeutic levels, increased the available thrombin activity, yet argatroban still inhibited this clot-associated thrombin with IC50 values in the low micromolar range. Scanning electron microscopy/morphometric analyses demonstrated that permeation with argatroban had no significant effects on clot structure. We conclude that argatroban is an effective inhibitor of thrombin bound to aged fibrin clots, in purified systems and in plasma clots, as well as in clots that have been treated with the thrombolytic agents streptokinase and alteplase. PMID- 9731065 TI - Lysis of plasma clots by urokinase-soluble urokinase receptor complexes. AB - Single-chain urokinase plasminogen activator (scuPA), the unique form secreted by cells, expresses little intrinsic plasminogen activator activity. scuPA can be activated by proteolytic cleavage to form a two-chain enzyme (tcuPA), which is susceptible to inhibition by plasminogen activator inhibitor type I (PAI-1). scuPA is also activated when it binds to its cellular receptor (uPAR), in which case the protein remains as a single chain molecule with less susceptibility to PAIs. Fibrin clots are invested with PAI-1 derived from plasma and from activated platelets. Therefore, we compared the fibrinolytic activity of complexes between scuPA and recombinant soluble uPAR (suPAR) to that of scuPA, tcuPA, and tcuPA/suPAR complexes. scuPA/suPAR complexes mediated the lysis of plasma-derived fibrin clots 14-fold more extensively than did equimolar concentrations of scuPA and threefold more extensively than did tcuPA or tcuPA/suPAR, respectively. The enhanced catalytic activity of scuPA/suPAR required that all three domains of the receptor be present, correlated with its PAI-1 resistance, was not dependent on fibrin alone, and required a plasma cofactor that was identified as IgG. Human IgG bound specifically to suPAR and scuPA/suPAR as determined by using affinity chromatography and immunoprecipitation. Plasma depleted of IgG lost most of its capacity to promote the fibrinolytic activity of scuPA/suPAR, and the activity of the complex was restored by adding plasma concentrations of purified IgG. These studies indicate that scuPA/suPAR can function as a plasminogen activator in a physiological milieu. PMID- 9731066 TI - Elevated levels of serum-soluble CD14 in human immunodeficiency virus type 1 (HIV 1) infection: correlation to disease progression and clinical events. AB - Soluble (s) CD14, a marker for monocyte/macrophage activation and a mediator of bacterial lipopolysaccharide (LPS) action, was elevated in serum from human immunodeficiency virus type 1 (HIV- 1)-infected individuals (n = 92) compared with seronegative controls. The highest levels were found in patients with advanced clinical and immunological disease. Patients with ongoing clinical events had significantly higher sCD14 levels than symptomatic HIV-1-infected individuals without clinical events, with especially elevated levels in patients infected with Mycobacterium avium complex (MAC). On longitudinal testing of patients (n = 26) with less than 100 x 10(6) CD4 lymphocytes/L at baseline, we found that increasing sCD14 serum concentrations per time unit were associated with death, whereas no differences in CD4 cell number decrease were found between survivors and nonsurvivors. In vitro studies showed that HIV-1 glycoprotein 120 and purified protein derivative (PPD) from M avium (MAC-PPD) stimulated normal monocytes to release sCD14. Furthermore, MAC-PPD induced tumor necrosis factor (TNF) release from monocytes through interactions with CD14 and, importantly, the addition of sCD14 enhanced this MAC-PPD stimulatory effect. Our findings suggest that the CD14 molecule may be involved in the immunopathogenesis of HIV-1 infection, and it is conceivable that serial determination of sCD14 may give useful predictive information concerning disease progression and survival in HIV 1-infected patients. PMID- 9731067 TI - The induction of nitric oxide by interleukin-12 and tumor necrosis factor-alpha in human natural killer cells: relationship with the regulation of lytic activity. AB - We have investigated the interleukin-12 (IL-12) and tumor necrosis factor-alpha (TNFalpha)-induced regulation of human natural killer (NK) cell function and their relationship with nitric oxide (NO) generation. We demonstrate that both cytokines were efficient to trigger the transcription of the inducible nitric oxide synthase (iNOS) mRNA, as detected by reverse transcriptase-polymerase chain reaction (RT-PCR). Western blot analysis and intracytoplasmic fluorescence showed that iNOS protein was also induced by both cytokines. However, our data indicate that NO does not play a significant role in the effector phase of the cytotoxic activity mediated by NK-stimulated cells, inasmuch as the lytic activity was not affected in the presence of specific NO synthase inhibitors. When aminoguanidine (AMG), an inhibitor of iNOS, was added during the afferent phase of NK stimulation with IL-12 and TNFalpha, a subsequent increase in the lytic potential of the effector cells towards the NK-sensitive target cells (K562) and lymphokine activated killer (LAK) target cells (Daudi) was observed. Conversely, the addition of chemical NO donors during the afferent step resulted in a dose dependent inhibition of the NK and LAK cytotoxicity. Our data suggest that the enhancement of NK-cell cytotoxic activity resulting from iNOS inhibition may be correlated, at least in part, to an increase in interferon-gamma production and granzyme B expression. PMID- 9731068 TI - Treatment of B-cell lymphoma with chimeric IgG and single-chain Fv antibody interleukin-2 fusion proteins. AB - Anti-idiotype (Id) antibodies (Abs) have been shown to be effective in treatment of B-cell lymphoma in animal models and in clinical trials. The combination of interleukin-2 (IL-2) can augment the therapeutic effect of anti-Id Abs. To further improve the power of the combined therapy, a monoclonal anti-Id Ab, S5A8, specifically recognizing a murine B-cell lymphoma 38C13, was genetically modified to contain the IL-2 domain and thus use the unique targeting ability of Abs to direct IL-2 to the tumor site. Two forms of the anti-Id-IL-2 fusion proteins were constructed: one configuration consisting of mouse-human chimeric IgG (chS5A8-IL 2) and the other containing only the variable light (VL) and variable heavy (VH) Ab domains covalently connected by a peptide linker (scFvS5A8-IL-2). Both forms of the anti-Id-IL-2 fusion proteins retained IL-2 biological activities and were equivalent in potentiating tumor cell lysis in vitro. In contrast, the antigen binding ability of scFvS5A8-IL-2 was 30- to 40-fold lower than that of the bivalent chS5A8-IL-2. Pharmacokinetic analysis showed that scFvS5A8-IL-2 was eliminated about 20 times faster than chS5A8-IL-2. Finally, it was shown that chS5A8-IL-2 was very proficient in inhibiting 38C13 tumor growth in vivo, more effectively than a combined therapy with anti-Id Abs and IL-2, whereas scFvS5A8 IL-2 did not show any therapeutic effect. These results demonstrate that the anti Id-IL-2 fusion protein represents a potent reagent for treatment for B-cell lymphoma and that the intact IgG fusion protein is far more effective than its single-chain counterpart. PMID- 9731069 TI - Allelotype analysis of adult T-cell leukemia. AB - Allelotype analysis of adult T-cell leukemia (ATL) was undertaken for the first time to identify chromosomal loci relevant to the development of acute/lymphomatous ATL. Loss of heterozygosity (LOH) was screened using 94 highly polymorphic microsatellite markers, distributed among all nonacrocentric, autosomal chromosomes. In each of the 22 cases, DNA obtained from their leukemic cells in acute/lymphomatous phase was compared with their constitutional DNA from mononuclear cells in chronic or remission phase. Allelic losses of at least on one chromosome arm occurred in 91% of the cases (20 individuals). Among 39 chromosome arms, allelic losses were observed on 31 arms at least for one sample. A high frequency of allelic loss (>30%) was seen on chromosome arms 6q (41%) and 17p (48%). The mean fractional allelic loss (FAL) was 0.109. These findings suggest that a novel tumor suppressor gene on chromosome arm 6q, as well as the p53 gene on chromosome arm 17p, probably have an important role in the development of acute/lymphomatous ATL. PMID- 9731070 TI - Acute mixed lineage leukemia with an inv(8)(p11q13) resulting in fusion of the genes for MOZ and TIF2. AB - Chromosomal abnormalities in acute leukemia have led to the discovery of many genes involved in normal hematopoiesis and in malignant transformation. We have identified the fusion partners in an inv(8)(p11q13) from a patient with acute mixed lineage leukemia. We show by fluorescence in situ hybridization (FISH) analysis, Southern blotting, and reverse transcriptase-polymerase chain reaction (RT-PCR) that the genes for MOZ, monocytic leukemia zinc finger protein, and TIF2, transcriptional intermediary factor 2, are involved in the inv(8)(p11q13). We demonstrate that the inversion creates a fusion between the 5' end of MOZ mRNA and the 3' end of TIF2 mRNA maintaining the translational frame of the protein. The predicted fusion protein contains the zinc finger domains, the nuclear localization domains, the histone acetyltransferase (HAT) domain, and a portion of the acidic domain of MOZ, coupled to the CREB-binding protein (CBP) interaction domain and the activation domains of TIF2. The breakpoint is distinct from the breakpoint in the t(8;16)(p11;p13) translocation in acute monocytic leukemia with erythrophagocytosis that fuses MOZ with CBP. The reciprocal TIF2 MOZ fusion gene is not expressed, perhaps as a result of a deletion near the chromosome 8 centromere. The MOZ-TIF2 fusion is one of a new family of chromosomal rearrangements that associate HAT activity, transcriptional coactivation, and acute leukemia. PMID- 9731071 TI - Characterization of siglec-5, a novel glycoprotein expressed on myeloid cells related to CD33. AB - We describe the characterization of siglec-5 (sialic acid-binding Ig-like lectin 5), a novel transmembrane member of the immunoglobulin superfamily, highly related to the myeloid antigen, CD33. A full-length cDNA encoding siglec-5 was isolated from a human activated monocyte cDNA library. Sequencing predicted that siglec-5 contains four extracellular immunoglobulin-like domains, the N-terminal two of which are 57% identical to the corresponding region of CD33. The cytoplasmic tail is also related to that of CD33, containing two tyrosine residues embodied in immunoreceptor tyrosine-based inhibitory motif-like motifs. The siglec-5 gene was shown to map to chromosome 19q13.41-43, closely linked to the CD33 gene. When siglec-5 was expressed on COS cells or as a recombinant protein fused to the Fc region of human IgG1, it was able to mediate sialic acid dependent binding to human erythrocytes and soluble glycoconjugates, suggesting that it may be involved in cell-cell interactions. By using specific antibodies, siglec-5 was found to have an expression pattern distinct from that of CD33, being present at relatively high levels on neutrophils but absent from leukemic cell lines representing early stages of myelomonocytic differentiation. Western blot analysis of neutrophil lysates indicated that siglec-5 exists as a disulfide linked dimer of approximately 140 kD. PMID- 9731072 TI - Activation of the small GTPase rap1 in human neutrophils. AB - The small GTPase Rap1 is highly expressed in human neutrophils, but its function is largely unknown. Using the Rap1-binding domain of RalGDS (RalGDS-RBD) as an activation-specific probe for Rap1, we have investigated the regulation of Rap1 activity in primary human neutrophils. We found that a variety of stimuli involved in neutrophil activation, including fMet-Leu-Phe (fMLP), platelet activating factor (PAF), granulocyte-macrophage colony-stimulating factor (GM CSF), and IgG-coated particles, induce a rapid and transient Rap1 activation. In addition, we found that Rap1 is normally activated in neutrophils from chronic granulomatous disease patients that lack cytochrome b558 or p47phox and have a defective NADPH oxidase system. From these results we conclude that in neutrophils Rap1 is activated independently of respiratory burst induction. Finally, we found that Rap1 is activated by both the Ca2+ ionophore ionomycin and the phorbol ester 12-O-tetradecanoylphorbol 13-acetate (TPA), indicating that phospholipase C (PLC) activation leading to elevated levels of intracellular free Ca2+ and diacylglycerol (DAG) can mediate Rap1 activation. However, inhibition of PLC and Ca2+ depletion only marginally affected fMLP-induced Rap1 activation, suggesting that additional pathways may control Rap1 activation. PMID- 9731073 TI - Is hemoglobin instability important in the interaction between hemoglobin E and beta thalassemia? AB - Hemoglobin E (HbE; alpha2beta226glu-lys), globally the commonest hemoglobin variant, is synthesized at a slightly reduced rate and has a homozygous phenotype similar to heterozygous beta thalassemia. Yet, when it is inherited together with a beta thalassemia allele, the resulting condition, HbE/beta thalassemia, is sometimes characterized by a severe, transfusion-dependent thalassemia major. The severity of this interaction has not been explained. We have explored the possibility that it may reflect the instability of HbE consequent upon globin chain imbalance imposed by the beta thalassemia allele. Time-course and pulse chase globin chain synthesis studies at 37 degrees C on peripheral blood and bone marrow suggest that hemoglobin instability is not significant in steady-state HbE/beta thalassemia; this is confirmed by density-gradient centrifugation studies that show no decrease in HbE levels relative to HbA as HbE/beta+ thalassemia red blood cells age. Globin binding to membranes was assessed and only alpha globin chains were found, in contrast to other unstable hemoglobins in which both alpha and beta chains were present. However, in experiments performed on blood from HbE/beta thalassemics in the temperature range 39 degrees C to 41 degrees C, there was evidence of instability of HbE, a finding that was also observed in homozygous HbE. These findings suggest that the phenotype of HbE/beta thalassemia is primarily the result of the interaction of two beta thalassemia alleles; however, hemoglobin instability may be important during febrile episodes, contributing to worsening anemia. PMID- 9731074 TI - Arg89Cys substitution results in very low membrane expression of the Duffy antigen/receptor for chemokines in Fy(x) individuals. AB - The Duffy (FY) blood group antigens are carried by the DARC glycoprotein, a widely expressed chemokine receptor. The molecular basis of the Fya/Fyb and Fy(a b-) polymorphisms has been clarified, but little is known about the Fyx antigen and the FY*X allele associated with weak expression of Fyb, Fy3, Fy5, and Fy6 antigens. We analyzed here the structure and expression of the FY gene in 4 Fy(a bweak) individuals. As compared with Fy(a-b+) controls, the Fy(a-bweak) red blood cell membranes contained residual amount of DARC polypeptide and these cells were poorly bound by anti-Fy antibodies and chemokines. The FY gene from Fy(a-b+) and Fy(a-bweak) individuals differed by one substitution, C286T. The resulting Arg89Cys amino acid change reduced the binding of anti-Fy antibodies and chemokines to DARC transfectants. We concluded that the Fybweak donors carried the FY*X allele at the FY locus and that the Fyx antigen corresponds to highly reduced expression of a grossly normal Fyb polypeptide caused by the Arg89Cys substitution. Because FY is a single copy gene, this defect should also affect DARC expression in nonerythroid cells. Because the Fyx phenotype is not associated with apparent clinical consequences, we discussed these findings in the light of the putative roles of DARC in various tissues. Finally, we developed a Fyx DNA typing assay that should be useful for genetic studies and clinical transfusion medicine. PMID- 9731075 TI - The G185R mutation disrupts function of the iron transporter Nramp2. AB - Microcytic anemia (mk) mice and Belgrade (b) rats have severe iron deficiency anemia due to defects in intestinal iron transport and erythroid iron utilization. Both animal mutants carry the same missense mutation in Nramp2, the first mammalian iron transporter to be identified. This mutation, in which glycine 185 is changed to arginine (G185R), occurs within predicted transmembrane domain 4 of the protein. We have performed site-directed mutagenesis of murine Nramp2, focusing on amino acids of transmembrane domain 4 that are highly conserved among Nramp-like proteins. We have expressed each mutant form in transfected cells and examined iron transport function, subcellular localization, and protein amounts. All tested forms of Nramp2 localize to the plasma membrane and to transferrin-containing endosomes. Most transmembrane domain 4 mutations affect the amount of protein detected and consequently show diminished iron transport. The G185R mutation, however, causes near total loss of Nramp2 function that cannot be fully explained by a decreased amount of protein, indicating that G185R disrupts iron transport through an alteration in the function of Nramp2, rather than degradation of the protein. PMID- 9731076 TI - Hepatitis G virus RNA and hepatitis G virus-E2 antibodies in Dutch hemophilia patients in relation to transfusion history. AB - The prevalence of hepatitis G virus (HGV)-RNA and HGV-E2 antibodies was studied in a cohort of Dutch hemophilia patients in relation to clotting products used, age, and coinfection with hepatitis C. Between 1991 and 1995, blood samples were taken from 294 patients with hemophilia A, B, or von Willebrand disease. From each patient one fresh frozen sample was tested for HGV cDNA polymerase chain reaction (PCR) and HCV cDNA PCR. Alanine aminotransferase (ALT) tests were performed on plasma samples of all patients. The presence of HGV-E2 antibodies was tested on plasma samples from a subset of 169 patients representing all age groups. Based on the origin and viral safety of the products used, three subgroups of patients were distinguished. Group A: patients who used viral noninactivated factors derived from small and large donor pools; group B: patients who used factors prepared with inadequate viral inactivation techniques derived from small and large donor pools; and group C: patients treated only with optimally viral inactivated large pool clotting factor or recombinant clotting factor concentrate. The prevalence of HGV-RNA was 18%. In group A patients the prevalence was 71%, in group B 50%, and in group C 6%. When related to age, the highest prevalence of HGV-RNA (35%) was seen in patients born between 1980 and 1989. The prevalence of HGV-E2 antibodies increased with age. Of HGV-RNA-negative patients born before 1950, 96% tested positive. HGV viremia did not affect ALT levels, neither in HCV-RNA positive nor in HCV-RNA negative patients. HGV infection is frequently seen in patients with hemophilia. In older age groups a lower rate of HGV-RNA positivity is seen coinciding with a higher rate of antienvelope antibodies. PMID- 9731077 TI - Evidence that CD31, CD49b, and CD62L are immunodominant minor histocompatibility antigens in HLA identical sibling bone marrow transplants. AB - Despite complete matching of siblings for the HLA loci, after bone marrow transplantation (BMT), approximately 20% develop graft-versus-host disease (GVHD). This is presumably due to incompatibility of minor histocompatibility antigens (mHa). We investigated the polymorphisms of 14 adhesion molecules (CD2, CD28, CD31, CD34, CD36, CD42, CD44, CD48, CD49b, CD54, CD62L, CD86, CD102, and CD106) in Japanese subjects and their association with the occurrence of GVHD after allogeneic HLA identical BMT. Six molecules (CD2, CD31, CD42, CD49b, CD54, and CD62L), which were found to be polymorphic, were then examined in 118 HLA identical sibling donors and recipients who had undergone BMT. Association of the incompatibility of the polymorphic molecules with the presence or absence of GVHD was examined. In these six, we observed a significant correlation between acute GVHD and the compatibility of CD31 (codons 563/670) (Pcorrected = .018), and CD31 (codons 563/670) + CD62L (Pcorrected = .018) in patients with the HLA-B44-like superfamily. In patients with the HLA-A3-like superfamily, the compatibility of CD62L (Pcorrected = .03) and CD62L + CD49b (P = . 004, Pcorrected = .078) was associated with acute GVHD. Therefore, CD31, CD49b, and CD62L might be candidates for immunodominant mHa. PMID- 9731078 TI - Involvement of donor T-cell cytotoxic effector mechanisms in preventing allogeneic marrow graft rejection. AB - Donor CD8 cells play a pivotal role in preventing allogeneic marrow graft rejection, possibly by generating cytotoxic effectors needed to eliminate recipient T cells remaining after the pretransplant conditioning regimen or by producing cytokines needed to support the growth and differentiation of hematopoietic stem cells. In the present study, we assessed the role of donor T cell cytotoxic effector function as a mechanism for eliminating recipient CD8 cells that cause marrow graft rejection in mice. The ability to prevent rejection was minimally affected by the presence of a defect in Fas ligand binding or by the absence of granzyme B but was severely affected by the absence of perforin. Doubly mutant perforin-deficient, Fas ligand-defective CD8 cells were completely unable to prevent rejection. Our results indicate first that recipient CD8 effectors responsible for causing marrow graft rejection are sensitive to cytotoxicity mediated by both perforin- and Fas-ligand-dependent mechanisms, and second that donor T cells must have at least one functional cytotoxic mechanism to prevent allogeneic marrow graft rejection. PMID- 9731080 TI - Difficulties in determining prophylactic transfusion thresholds of platelets in leukemia patients. PMID- 9731079 TI - Detection of antibodies to Fasciola and Anisakis in Japanese patients with intravascular lymphomatosis. PMID- 9731081 TI - The Italian experience on interferon as maintenance treatment in multiple myeloma: ten years after. PMID- 9731082 TI - Kaposi's sarcoma-associated herpesvirus is not detected with immunosuppression in multiple myeloma. PMID- 9731083 TI - Cytoplasmic interleukin-4 (IL-4) and surface IL-4 receptor expression in patients with B-cell lymphocytic leukemia. PMID- 9731084 TI - Primary role of the liver in thrombopoietin production shown by tissue-specific knockout. PMID- 9731085 TI - The pathway of exocytosis in human platelets. PMID- 9731086 TI - The prevalence of cardiac valvular insufficiency assessed by transthoracic echocardiography in obese patients treated with appetite-suppressant drugs. AB - BACKGROUND: After case reports of cardiac-valve abnormalities related to the use of appetite suppressants were published, we undertook a study to determine the prevalence of the problem using transthoracic echocardiography. METHODS: We examined patients who had taken dexfenfluramine alone, dexfenfluramine and phentermine, or fenfluramine and phentermine for various periods. We enrolled obese patients who had taken or were taking these agents during open-label trials from January 1994 through August 1997. We also recruited subjects who had not taken appetite suppressants and who were matched to the patients for sex, height, and pretreatment age and body-mass index. The presence of cardiac-valve abnormalities, defined by the Food and Drug Administration and Centers for Disease Control and Prevention as at least mild aortic-valve or moderate mitral valve insufficiency, was determined independently by at least two cardiologists. Multivariate logistic-regression analysis was used to identify factors associated with cardiac-valve abnormalities. RESULTS: Echocardiograms were available for 257 patients and 239 control subjects. The association between the use of any appetite suppressant and cardiac-valve abnormalities was analyzed in a final matched group of 233 pairs of patients and controls. A total of 1.3 percent of the controls (3 of 233) and 22.7 percent of the patients (53 of 233) met the case definition for cardiac-valve abnormalities (odds ratio, 22.6; 95 percent confidence interval, 7.1 to 114.2; P<0.001). The odds ratio for such cardiac valve abnormalities was 12.7 (95 percent confidence interval, 2.9 to 56.4) with the use of dexfenfluramine alone, 24.5 (5.9 to 102.2) with the use of dexfenfluramine and phentermine, and 26.3 (7.9 to 87.1) with the use of fenfluramine and phentermine. CONCLUSIONS: Obese patients who took fenfluramine and phentermine, dexfenfluramine alone, or dexfenfluramine and phentermine had a significantly higher prevalence of cardiac valvular insufficiency than a matched group of control subjects. PMID- 9731087 TI - A population-based study of appetite-suppressant drugs and the risk of cardiac valve regurgitation. AB - BACKGROUND: Recent case reports suggest that a combination of the appetite suppressants fenfluramine and phentermine is associated with an increased risk of cardiac-valve regurgitation. There are also reports of valvular disorders in persons taking fenfluramine or dexfenfluramine alone, particularly for more than three months. METHODS: We conducted a population-based follow-up study and a nested case-control analysis of 6532 subjects who received dexfenfluramine, 2371 who received fenfluramine, and 862 who received phentermine to assess the risk of a subsequent clinical diagnosis of a valvular disorder of uncertain origin. For comparison, we identified a group of 9281 obese subjects who had not taken appetite suppressants who were matched to the treated subjects for age, sex, and weight. All subjects were free of diagnosed cardiovascular disease at the start of follow-up. The average duration of follow-up for all subjects was about four years. RESULTS: There were 11 cases of newly diagnosed idiopathic valvular disorders, 5 after the use of dexfenfluramine and 6 after the use of fenfluramine. There were six cases of aortic regurgitation, two cases of mitral regurgitation, and three cases of combined aortic and mitral regurgitation. There were no cases of idiopathic cardiac-valve abnormalities among the subjects who had not taken appetite suppressants or among those who took only phentermine. The five-year cumulative incidence of idiopathic cardiac-valve disorders was 0 per 10,000 subjects among those who had not taken appetite suppressants (95 percent confidence interval, 0 to 15.4) and among those who took phentermine alone (95 percent confidence interval, 0 to 76.6), 7.1 per 10,000 subjects among those who took either fenfluramine or dexfenfluramine for less than four months (95 percent confidence interval, 3.6 to 17.8; P=0.02 for the comparison with subjects who had not taken appetite suppressants), and 35.0 per 10,000 subjects among those who received either of these medications for four or more months (95 percent confidence interval, 16.4 to 76.2; P<0.001). CONCLUSIONS: The use of fenfluramine or dexfenfluramine, particularly for four months or longer, is associated with an increased risk of newly diagnosed cardiac-valve disorders, particularly aortic regurgitation. PMID- 9731088 TI - An assessment of heart-valve abnormalities in obese patients taking dexfenfluramine, sustained-release dexfenfluramine, or placebo. Sustained-Release Dexfenfluramine Study Group. AB - BACKGROUND: The appetite-suppressant drug fenfluramine, usually given in combination with phentermine, has been reported to be associated with cardiac valvular regurgitation. Concern has been raised that the d-enantiomer of fenfluramine, dexfenfluramine, may also cause this problem. We were able to study the question by modifying an ongoing trial comparing dexfenfluramine with regular dexfenfluramine and placebo. METHODS: We modified our randomized, double-blind, placebo-controlled study of dexfenfluramine to include echocardiographic examinations of 1072 overweight patients within a median of one month after the discontinuation of treatment. The patients (approximately 80 percent of whom were women) had been randomly assigned to receive dexfenfluramine (366 patients), investigational sustained-release dexfenfluramine (352 patients), or placebo (354 patients). The average duration of treatment was 71 to 72 days in each of the three groups. Echocardiograms were assessed in a blinded fashion. RESULTS: When all degrees of valvular regurgitation were considered and when the two dexfenfluramine groups were combined, there was a higher prevalence of any degree of aortic regurgitation (17.0 percent vs. 11.8 percent, P=0.03) and any degree of mitral regurgitation (61.4 percent vs. 54.4 percent, P=0.01) in the active treatment groups than in the placebo group. These differences were primarily due to a higher prevalence of physiologic, trace, or mild regurgitation. Analyses that used the criteria of the Food and Drug Administration for aortic regurgitation of mild or greater severity and mitral regurgitation of moderate or greater severity found no statistically significant difference among the groups (P=0.14 to 0.75). These analyses showed that aortic regurgitation of mild or greater severity occurred in 5.0 percent of the patients in the dexfenfluramine group, 5.8 percent of those in the sustained-release dexfenfluramine group, 5.4 percent of those in the two active-treatment groups combined, and 3.6 percent of those in the placebo group. Mitral regurgitation of moderate or greater severity occurred in 1.7, 1.8, 1.8, and 1.2 percent, respectively. Aortic regurgitation of mild or greater severity, mitral regurgitation of moderate or greater severity, or both occurred in 6.5 percent, 7.3 percent, 6.9 percent, and 4.5 percent, respectively. CONCLUSIONS: The increased prevalence of aortic and mitral regurgitation in patients treated with dexfenfluramine was small, and the degree of regurgitation was usually classified as physiologic, trace, or mild. However, the duration of therapy was short, and whether therapy of longer duration would yield the same or different results is not known. PMID- 9731089 TI - Endogenous hormones and the risk of hip and vertebral fractures among older women. Study of Osteoporotic Fractures Research Group. AB - BACKGROUND AND METHODS: In postmenopausal women, the serum concentrations of endogenous sex hormones and vitamin D might influence the risk of hip and vertebral fractures. In a study of a cohort of women 65 years of age or older, we compared the serum hormone concentrations at base line in 133 women who subsequently had hip fractures and 138 women who subsequently had vertebral fractures with those in randomly selected control women from the same cohort. Women who were taking estrogen were excluded. The results were adjusted for age and weight. RESULTS: The women with undetectable serum estradiol concentrations (<5 pg per milliliter [18 pmol per liter]) had a relative risk of 2.5 for subsequent hip fracture (95 percent confidence interval, 1.4 to 4.6) and subsequent vertebral fracture (95 percent confidence interval, 1.4 to 4.2), as compared with the women with detectable serum estradiol concentrations. Serum concentrations of sex hormone-binding globulin that were 1.0 microg per deciliter (34.7 nmol per liter) or higher were associated with a relative risk of 2.0 for hip fracture (95 percent confidence interval, 1.1 to 3.9) and 2.3 for vertebral fracture (95 percent confidence interval, 1.2 to 4.4). Women with both undetectable serum estradiol concentrations and serum sex hormone-binding globulin concentrations of 1 microg per deciliter or more had a relative risk of 6.9 for hip fracture (95 percent confidence interval, 1.5 to 32.0) and 7.9 for vertebral fracture (95 percent confidence interval, 2.2 to 28.0). For those with low serum 1,25-dihydroxyvitamin D concentrations (< or =23 pg per milliliter [55 pmol per liter]), the risk of hip fracture increased by a factor of 2.1 (95 percent confidence interval, 1.2 to 3.5). CONCLUSIONS: Postmenopausal women with undetectable serum estradiol concentrations and high serum concentrations of sex hormone-binding globulin have an increased risk of hip and vertebral fracture. PMID- 9731091 TI - A Medical Mystery. PMID- 9731090 TI - Chlamydia trachomatis infections in female military recruits. AB - BACKGROUND: Asymptomatic genital Chlamydia trachomatis infections in women can lead to pelvic inflammatory disease, infertility, and ectopic pregnancy. To design a chlamydia-control program, we conducted a large survey of women in the U.S. military. METHODS: From January 1996 through December 1997, urine samples from 13,204 new female U.S. Army recruits from 50 states were screened by ligase chain reaction for C. trachomatis infection. Information on potential risk factors was obtained by questionnaire. With multivariate analysis, we identified criteria for a screening program. RESULTS: The overall prevalence of chlamydial infection was 9.2 percent, with a peak of 12.2 percent among the 17-year-old recruits. The prevalence was 15 percent or more among the recruits from five southern states. The following risk factors were independently associated with chlamydial infection: having ever had vaginal sex (odds ratio for infection, 5.9), being 25 years of age or less (odds ratio, 3.0), being black (odds ratio, 3.4), having had more than one sex partner in the previous 90 days (odds ratio, 1.4), having had a new partner in the previous 90 days (odds ratio, 1.3), having had a partner in the previous 90 days who did not always use condoms (odds ratio, 1.4), and having ever had a sexually transmitted disease (odds ratio, 1.2). A screening program for subjects 25 years of age or less (87.9 percent of our sample) would have identified 95.3 percent of the infected women. CONCLUSIONS: Among female military recruits, the prevalence of chlamydial infection is high. A control program that screens female recruits who are 25 years old or younger with urine DNA-amplification assays has the potential to reduce infection, transmission, and the sequelae of chlamydial infection. PMID- 9731092 TI - Thrombopoietin. PMID- 9731094 TI - Appetite suppressants and valvular heart disease. PMID- 9731096 TI - Expanding efforts to prevent chlamydial infection. PMID- 9731095 TI - Estrogen and the risk of fracture--new data, new questions. PMID- 9731097 TI - Comparison of euro-collins solution, low-potassium dextran solution containing glucose, and ET-kyoto solution for lung preservation in an extracorporeal rat lung perfusion model. PMID- 9731098 TI - Effects of growth hormone and insulin-like growth factor-I on radiation enteritis. a comparative study. PMID- 9731099 TI - Healing of colonic anastomoses after immediate and delayed administration of 5 fluorouracil plus folinic acid. PMID- 9731100 TI - Inhibition of myointimal proliferation by octreotide in canine vein interposition grafts. PMID- 9731101 TI - Metabolism of superoxide dismutase during the adynamic ileus of endotoxemia. PMID- 9731102 TI - Effect of surgery on neutrophil leukotriene B4 generation and arachidonic acid content. PMID- 9731104 TI - Recognition of a gastroesophageal reflex in dogs and its role in lower esophageal sphincter competence. PMID- 9731103 TI - An ex vivo perfusion model to evaluate hyperacute rejection in a discordant pig to-human combination. PMID- 9731106 TI - Annexin V expression and membrane vesiculation during activation of leukemic cell lines. AB - We investigated annexin V expression and membrane vesiculation during activation of four leukemic cell lines (U937, HL60, HEL, and CMK11-5) in order to determine whether annexin V had a role in the coagulation abnormalities related to malignancy. After stimulation by tissue plasminogen activator, binding of a monoclonal anti-annexin V antibody to U937 cells and HL60 cells increased in comparison with binding to control cells. Stimulation with thrombin or lipopolysaccharide also induced such an increase, but U46619 did not. Following activation of U937 and HL60 cells with thrombin and lipopolysaccharide, microparticle formation increased. Tissue plasminogen activator caused an increase of microparticles in U937 cells, but not HL60 cells. On the other hand, CMK11-5 and HEL cells did not show any increase of microparticles. These results suggest that some agonists can potently stimulate expression of prothrombinase PMID- 9731107 TI - Beneficial effect of treatment with a monoclonal anti-tumor necrosis factor-alpha antibody on markers of coagulation and fibrinolysis in patients with active Crohn's disease. AB - Crohn's disease has frequently been associated with coagulation abnormalities, causing intravascular deposition of fibrin and local infarction which can subsequently compromise the gut mucosa. Also, arterial and venous thromboembolic complications of larger vessels appear to be associated with Crohn's disease. Coagulation activation in patients with Crohn's disease could be a result of increased serum and tissue levels of cytokines, as reported. We prospectively studied parameters of coagulation and fibrinolysis in 10 patients with active Crohn's disease, who were subsequently treated with a monoclonal anti-tumor necrosis factor-alpha (TNF) antibody. Ten consecutive patients with active Crohn's disease (CDAI > 150), not responding to a daily dose of at least 20 mg prednisolone, received a single infusion of human/mouse chimeric anti-TNF antibody cA2. All evaluable patients attained complete clinical and endoscopic PMID- 9731108 TI - Protective role of platelets in chronic (Balb/C) and acute (CBA/J) Plasmodium berghei murine malaria. AB - The role of blood platelets in the pathogenesis of malaria remains unclear. In this study we investigated the role of experimentally caused thrombocytopaenia in chronic (Balb/C) and acute (CBA/J) Plasmodium-berghei-induced murine malaria. A group of 30 Balb/C mice were rendered thrombocytopaenic by injection of anti mouse platelet serum given every 2 days from day 2 to day 8 after malaria infection. Compared with the control group, thrombocytopaenic mice showed a greater mortality. The Kaplan-Meier estimate of the risk of death was 4.37-fold higher in the thrombocytopaenic group. Parasitaemia and weight loss was in agreement with the protective effect of platelets. In the acute malaria model, the survival rate was less significant but the estimated risk of death was 1.7 fold higher in the treated group. These results suggesting a protective role of platelets in murine malaria are not in line with previous reports of the literature. Taken together our data suggest, however, that platelets, similarly to some inflammatory cytokines like TNF, play a dual role in the pathogenesis of malaria. Depending on experimental conditions, e.g. the time of onset of thrombocytopaenia, the beneficial role of platelets may outweigh the deleterious one. PMID- 9731109 TI - Acquired type 2a von Willebrand's disease: response to immunoglobulin infusion. AB - A 75-year-old male presented with new symptoms of a bleeding diathesis associated with a decline in the functional activity of von Willebrand factor (type 2a von Willebrand's disease). Replacement therapy was ineffective and he was subsequently treated with intravenous immunoglobulin (IvIg). IvIg not only caused symptomatic improvement but was shown to transiently restore the depleted von Willebrand factor intermediate and high molecular weight multimers. Subsequent periodic IvIg infusions have been used successfully to treat bleeding complications in this patient over the past 3 years. A secondary cause for the acquired von Willebrand's disease has not been identified. PMID- 9731110 TI - A highly specific functional test for factor V leiden: A modified tissue factor assay for activated protein C resistance. AB - We compared the sensitivity and specificity of a tissue factor-based assay (FVR) with the addition of a phospholipid/silica preparation, to the commercially available aPTT-based method, APCR (CoatestTM), and a modified aPTT-based method (APCM) which utilized factor V-depleted plasma, for the detection of the factor V Leiden mutation. A total of 110 patients were included in this study. This included 32 patients on coumadin therapy, 7 patients on heparin therapy, 5 patients on both anticoagulants therapy, and 24 patients who were positive for anticardiolipin antibody (ACL) and/or lupus inhibitor (LI). Our data demonstrate that the FVR is not affected by anticoagulation treatment or ACL/LI antibodies, whereas in the APCR method, 33 patients cannot be determined either due to the anticoagulant therapy or presence of the ACL and/or LI. With the APCM method, the clotting endpoint could not be determined in 1 patient due to the presence of a strong LI. The additional phospholipid/silica material utilized in the FVR enhanced the APC degradation of factor Va and therefore sharpened the demarcation between the factor V Leiden-positive and -negative patients. The sensitivity for the APCR, APCM and FVR was 42, 97 and 100% respectively. The specificity for the APCR, APCM and FVR was 94, 96 and 100% respectively. PMID- 9731111 TI - Diurnal variation in serum remnant-like lipoproteins, platelet aggregation and fibrinolysis in healthy volunteers. AB - Postprandial triglyceridemia and 'remnantemia' may better reflect the atherosclerotic risk than triglyceride (TG) levels in the fasting state. Recently, a new method was developed based on a monoclonal antibody recognizing an epitope distal to the carboxyl end of apo B48 which allows easy measurement of remnant-like lipoproteins (RLP). This study was performed in order to investigate RLP response to a standardized fat meal and establish a normal diurnal pattern of RLP in blood and compare it to platelet aggregation and fibrinolysis in healthy young men. We investigated 7 male volunteers (age range 18-23 years) who received a standardized fat meal (Othsuka Pharmaceutical Company, Japan) containing 32.9% lipids, 2.5% protein, 2.5% carbohydrate, 0.3% calcium and 0.1% phospholipids, and 74 mg/100 g cholesterol (C) at 7:30. The energetic value of this cocktail was 341 kcal/100 g. Area under curve (AUC) responses in TG, RLP-TG and RLP-C after the meal were as follows: for TG 28.66 +/- 8.94; for RLP-TG 17.54 +/- 5.55; for RLP-C 1.27 +/- 0.42 mg x dl-1 x h-1. These responses were correlated to each other. Surprisingly, collagen-induced platelet aggregation in whole blood was negatively related to RLP-C AUC. Fluctuation patterns of TG, RLP-TG and RLP-C concentrations during the day were remarkably similar, peaking in this particular group of subjects at 10:00-12:00 and at about 23:00, whereas cholesterol was decreasing late in the night and very early in the morning. This pattern was different from those of platelet aggregation and fibrinolysis parameters. PMID- 9731115 TI - Inflammation measurement and immunocharacterization of cell proliferation in an experimental model of proliferative vitreoretinopathy. AB - An experimental model of proliferative vitreoretinopathy was developed in the rabbit eye by injecting a solution of human platelet-rich plasma. In this model we evaluated the progression with time of intraocular inflammation and the rate and origin of cell proliferation. A sterile solution adjusted to 107 platelets was injected into the right eye of a total of 46 pigmented and 14 albino rabbits. Animals were sequentially sacrificed at days 7, 14, 21 and 1 month after injection. Clinical evaluation of vitreoretinal proliferation, using a classification in six grades, and of anterior segment inflammation assessed by a Laser Flare Meter, were done for 1 month after injection, before histopathological analysis. Eighty percent of eyes developed tractional retinal detachment in 1 month. Histopathology showed intense cell migration and proliferation in the area of the ciliary body, as early as the seventh day, then further increasing rapidly. Infiltrates were composed of cytokeratin- and vimentin-expressing cells. Abnormal expression of vimentin was also found in ciliary and retinal epithelia and in M?ller cells. Inflammation measured by the Laser Flare Meter was maximal at day 11 and then reached a plateau at significantly higher levels than controls. Albino rabbits showed significantly lower grades of proliferation, as compared to pigmented rabbits. This study thus clarified some characteristics of experimental vitreoretinal proliferations that that proved similar to those in human diseases, such as the involvement of ciliary body and retinal pigment epithelium, the existence of inflammatory reactions preceding cell proliferation and strong changes in intermediate filaments. This may provide a simple and valuable model for antiproliferative assays and shed some light on the pathogenesis of intraocular proliferative disorders. PMID- 9731114 TI - Upregulation of retinal vascular endothelial growth factor mRNAs in spontaneously diabetic rats without ophthalmoscopic retinopathy. A possible participation of advanced glycation end products in the development of the early phase of diabetic retinopathy. AB - Vascular endothelial growth factor (VEGF) has recently been shown to be involved in the pathogenesis of proliferative diabetic retinopathy. However, its involvement in the development of the early phase of diabetic retinopathy is not fully understood. In this study we investigated the retinal VEGF mRNA level in spontaneously diabetic Otsuka Long-Evans Tokushima fatty (OLETF) rats, a model of non-insulin-dependent diabetes, without overt retinopathy, using quantitative reverse-transcription polymerase chain reaction. The retinal VEGF mRNA level was 2.2 times higher (p < 0.0005) in OLETF rats than in control rats at the age of 60 weeks. Moreover, their retinal mRNA level was positively correlated with serum concentration of advanced glycation end products (AGEs) but not to serum glucose concentration. Furthermore, the peak latency of the oscillatory potentials in the electroretinogram, one of the most sensitive markers for the early phase of diabetic retinopathy, was significantly prolonged in OLETF rats (p < 0.05), being also correlated with the serum AGE concentration. The results thus suggest that AGEs, which are formed acceleratedly in diabetic conditions, are involved in the development of the early phase of diabetic retinopathy probably through the induction of retinal VEGF mRNAs. PMID- 9731116 TI - The glutathione content of retinal Muller (glial) cells: the effects of aging and of application of free-radical scavengers. AB - The dependence of intracellular glutathione (GSH), an important radical scavenger, on aging with or without externally applied Ginkgo biloba extract EGb 761, another established radical scavenger, was studied in guinea pig M?ller (retinal glial) cells by using the fluorescent dye monochlorobimane. The GSH content of freshly dissociated cells from untreated aged animals was significantly lower than that of young controls; most of this reduction was prevented by application of EGb 761. Culturing the cells in amino-acid-free caused a loss of up to 50% of the initial GSH content. When the culture medium contained 100 microM glutamate and 100 microM cystine, ongoing GSH synthesis counteracted the loss of GSH. The rates of net GSH synthesis were equal for the two groups of aged animals but significantly higher for cells from young controls. It is concluded that externally applied radical scavengers may enhance the protective glutathione 'reserve' of M?ller cells in cases of neuronal degeneration. PMID- 9731117 TI - Retinoic acid increases in the retina of the chick with form deprivation myopia. AB - We previously reported that expression of retinoic acid receptor beta increases in the sclera of the 2-week-old chick with form deprivation myopia (FDM) and that all-trans-retinoic acid (t-RA) influences proliferation and differentiation of scleral cells. The purpose of this study was to quantify t-RA in the retina of the chick with FDM and to investigate the role of t-RA in FDM in the chick. FDM was induced in 2-day-old chicks by placement of a translucent plastic goggle over one eye, with the contralateral eye used as a control. After 5 days, the chicks were sacrificed. t-RA was extracted from neural retina and served for high performance liquid chromatography analysis. 3H-t-RA was used for normalization. Pieces of the retinae from 5 eyes served as one sample. As a result, t-RA was 09.60 +/- 0.86 ng/eye (0.387 +/- 0.056 ng/mg protein) in the myopic retina was significantly higher than that in the control (p < 0.05, n = 7). These results demonstrate that t-RA increases in the retina within 5 days after visual deprivation. This finding suggests that t-RA may play a role in the metabolic changes in FDM. PMID- 9731118 TI - 20-Hz flicker stimulus can isolate the cone function in rat retina. AB - Cone electroretinograms (ERGs) are typically isolated in humans by flicker stimuli against rod-desensitizing adapting fields. To investigate the manner in which adapting-field luminance affects the cone ERGs, we recorded ERGs in normal albino Sprague-Dawley rats with flicker stimuli presented against adapting fields that ranged in luminance from to 1.75 log cd/m2. A flicker rate of 20 Hz was used to isolate the cone ERGs under all adaptation conditions. We found the amplitudes of cone ERGs to increase with increasing adapting-field luminance. These response characteristics are similar to human ERGs using 30-Hz flicker stimuli, which suggests that flicker stimuli are a useful technique to isolate the cone function in rats. PMID- 9731119 TI - Sialoglycoconjugates in primary and recurrent mixed tumors of lacrimal glands. AB - We examined the activity of Griffonia simplicifolia II agglutinin (GS2) before and after neuraminidase treatment in 2 cases of primary benign mixed tumor (BMT) and a case of recurrent mixed tumor (MT). Histologically, each type of MT is comprised of epithelial and mesenchymal components. GS2, which showed no binding to either structural component in the primary BMT, was specifically bound to the mesenchymal component in the recurrent MT. After neuraminidase treatment, there was a marked increase in GS2-binding levels in the epithelial component of the primary BMT and in both components of the recurrent MT. These results might reveal a prominent sialylation in the recurrent MT. PMID- 9731120 TI - Laminin heterogeneity around Schlemm's canal in normal humans and glaucoma patients. AB - BACKGROUND: The laminins play an important role in cell attachment to the extracellular matrix. This study was undertaken in order to investigate and to compare the presence of several laminin chains and isoforms in normal and glaucomatous trabecular meshwork. METHODS: Fresh-frozen trabeculectomy specimens from glaucoma patients and trabecular meshwork from human donor eyes were included for immunohistochemistry. We used antibodies against laminin 1 (EHS laminin/alpha1beta1gamma1), laminin 2 (merosin/alpha2,beta1gamma1), the subunits of alpha2 (M laminin), beta1 B1laminin), gamma1 (beta2laminin) and beta2 (s laminin) and gamma2 and the associated glycoprotein nidogen. RESULTS: For laminin 1, laminin 2, the subunits alpha2;, beta1, beta2, gamma1 and nidogen, the immunoreactivity of the juxtacanalicular zone underneath the inner wall of Schlemm's canal was always stronger than of the uveal and corneoscleral trabecular meshwork. Staining for laminin chain gamma2 could not be demonstrated in the outflow structures. Pattern and intensity of labelling for the laminins were not different between normal and glaucomatous eyes. DISCUSSION: Heterogeneity of the laminins suggests structural complexity of the juxtacanalicular region. However, the presence of laminin around Schlemm's canal seems to be unaffected by glaucoma and by age. This might indicate maintenance of basement membrane stability in the trabecular meshwork for life. PMID- 9731121 TI - Uveal mast cells are not required for rodent uveitis. AB - Uveal mast cells have previously been considered to be vital mediators of experimental uveitis. We extended the study of these cells to experimental melanin-induced uveitis (EMIU), a recently described clinically relevant model, and re-examined their role in endotoxin-induced uveitis (EIU) using genetically mast cell-depleted mice on a single background. EMIU was induced in Fischer 344 rats by immunisation with bovine ocular melanin (250 microgram. Animals were killed immediately, and on days 1 and 3 of clinical disease. Numbers of uveal mast cells and the percentage of degranulated cells were counted in whole-mount preparations. There was no significant change in either measure across the selected time points. To induce EIU, normal and mast cell-depleted DBA/2 mice were injected with Escherichia coli lipopolysaccharide (400 microgram). Cells infiltrating the eye 24 h after injection were quantified in 5 micrometer ocular cross-sections. Disease was not significantly reduced in the mast cell-depleted mutants. We conclude that uveal mast cells are not required for the development of EMIU or, in contrast to earlier work, EIU. PMID- 9731122 TI - Visual function and gene analysis in a family with Oguchi's disease. AB - A family with 1 case of retinitis pigmentosa (III-1) and 2 cases of Oguchi's disease (III-2, 3) was examined in terms of electrophysiology as well as molecular biology. The proband (III-3), a 42-year-old female, and 2 older brothers (III-1, 2, aged 52 and 45 years) and 2 unaffected members in the same family participated in this study. Corrected visual acuities of the individuals with Oguchi's disease (III-2, 3) were 1.2. On funduscopy, blood vessels stood out in relief against a metallic-appearing background and a Mizuo-Nakamura phenomenon was evident. Full-field electroretinograms (ERGs) recorded from the proband were indicative of rod dystrophy, but results of other electrophysiological examinations (multifocal ERG, pattern ERG and visual-evoked cortical potential recordings) were within normal limits. Patient III-1 had corrected visual acuities of RE 20 cm/m.m. and LE 30 cm/n.d., severe chorioretinal atrophy in both fundi, and full-field ERG revealed rod-cone dystrophy. Mutation of the arrestin gene (1147de1A) was detected in all 3 patients. Visual function in each patient coincides with that of retinitis pigmentosa or Oguchi's disease, respectively. PMID- 9731123 TI - Lens transmission by fluorophotometry after brachytherapy and thermotherapy of choroidal melanoma. AB - We studied the effect of transpupillary thermotherapy (TTT) by a diode laser at 810 nm combined with episcleral ruthenium-106 plaque treatment (106Ru) on lens transparency in patients with choroidal melanoma. Lens transmission of blue-green light was measured by fluorophotometry in 17 patients treated with 106Ru treatment and TTT (measured 0.36 years after treatment), 12 patients treated with 106Ru alone (measured 19 years after treatment) and 25 age-matched healthy controls. Differences in lens transmission were not significant between treated and untreated fellow eyes (p > 0.15) nor between patient and control eyes (p > 0.25). TTT of choroidal melanoma combined with 106Ru plaque irradiation did not have a significant effect on the lens transparency up to 6 years after treatment. PMID- 9731124 TI - RNs unsung heroes during ice storm '98. PMID- 9731125 TI - Nursing informatics for beginners. PMID- 9731126 TI - The computer as tutor. PMID- 9731127 TI - Same-day pediatric surgery. PMID- 9731128 TI - [McEwen's conceptual model in cardiac rehabilitation]. AB - Health motivation is the importance individuals place on their will to persevere, which consequently influences their choice of lifestyle. This article focuses on a relatively new conceptual model that explains cardiac patients' motivation when it comes to initiating and sustaining healthy lifestyle habits. Developed by McEwen in 1993, the Health Motivation Model proposes the following series of variables that influence motivation: previous knowledge, perceived severity, perceived susceptibility, perceived value of action, background variables, internal aids/hindrances, external aids/hindrances and the catalyst. If nurses understand these variables and analyze the patient's case study, they will be better to investigate the origin of cardiac patients' resistance to positive lifestyle changes. PMID- 9731129 TI - Can gerontological nursing survive? AB - By the year 2041, 11 million individuals 65 and over will reside in Canada. This drastic demographic change, coupled with the increasing acuity and complexity of the health care needs of seniors, creates an immense demand for nurses specialized in gerontology. But can gerontological nursing survive as a specialty and respond to this demand? PMID- 9731130 TI - Teaching patients INs and OUTs. AB - When patients don't understand how to care for their chronic illness, frequent hospitalization results. One example is the patient who is admitted to a medical nursing unit in end stage renal disease (ESRD), the common complications of which are hypervolemia, hypovolemia and associated electrolyte imbalances. To prevent further disease progression and frequent hospitalization, an accurate measurement of fluid intake and output is critical, as is the patient's ability to understand and take responsibility for his/her own care. PMID- 9731131 TI - Children of hope. AB - Young people are emerging as a new casualty of the troubles in Northern Ireland. As the marching season begins, Craig Kenny talks to nurses who are eager to promote tolerance so the younger generation can benefit from the peace agreement. PMID- 9731132 TI - A 2020 vision of the future. PMID- 9731133 TI - Fifty years young: going on strong. PMID- 9731134 TI - People with disability may be more visible in the community but they are still being passed by. PMID- 9731135 TI - 'Lunch-break' abortion service seeks to expand. PMID- 9731136 TI - Memories are made of this. PMID- 9731137 TI - Waiting list lengths show no signs of abating. PMID- 9731138 TI - Far too many of us reach for the bottle. PMID- 9731139 TI - Death rights. PMID- 9731140 TI - Catheter care--2. PMID- 9731141 TI - The sky's the limit. PMID- 9731142 TI - The mother of all questions. PMID- 9731143 TI - Under scrutiny. PMID- 9731144 TI - Make love, not warfarin. PMID- 9731145 TI - Planning for action. PMID- 9731146 TI - Evidence-based practice in pressure sore reduction. AB - A two-year project at Chase Farm Hospitals NHS Trust to promote evidence-based practice in pressure sore prevention resulted in a steady reduction in the number of hospital-acquired pressure sores. The pooling of resources by a wide variety of staff with a diverse range of knowledge and experience resulted in a truly multiprofessional project. The knowledge that working together to implement change can succeed strengthens the chances of success in future work and has brought direct benefits to patient care. PMID- 9731147 TI - Management of suprapubic catheters. PMID- 9731148 TI - Clinical supervision and practice nurses. AB - practice nurses need clinical supervision to help them keep abreast of their expanding primary care role. Ali Farquharson, Gail Trotter and Sheila Nimmo report on a project that set out to provide a support model. PMID- 9731149 TI - Extended mental health service improves client well-being. PMID- 9731151 TI - Preventing infection from indwelling catheters. PMID- 9731150 TI - Using structured reflection to improve nursing practice. PMID- 9731152 TI - Catheters: making the right choice. PMID- 9731153 TI - Prevention is better than cure. PMID- 9731154 TI - Setting up an allergy clinic. PMID- 9731155 TI - Allergen avoidance. PMID- 9731156 TI - Towards a framework for women's health. AB - The health of women involves their emotional, social, cultural, spiritual and physical well-being and is influenced by social, political and economic factors, as well as by a woman's biology. This definition of women's health provides the conceptual framework for a relevant and effective approach to patient education, health promotion and disease prevention activities in women. Such activities must focus not only on diseases that are more common, more prevalent or more serious in women, but also on priority health issues identified by women themselves, the diversity of women, the determinants of health and above all, the impact of gender on health. These programs must also be based on knowledge resulting from gender sensitive, bias-free health research. The traditional oppression and disempowerment of women must also be addressed at both personal and societal levels, thus broadening our approach so that we become social as well as health activists. PMID- 9731157 TI - Speaking of illness: issues of first generation Canadian women--implications for patient education and counseling. AB - Based upon my research with first generation Canadian women who have come from Asia, I discuss the issues in women's lives that influence how they manage a chronic illness. I argue that women's relations with health care providers, the organization of the health care system, and structural constraints, especially for those women in the lower echelons of the labour force, profoundly influence the illness experience. Taking this, as the starting point, I proceed to examine the implications for education and counseling. I propose a model for counseling that begins with a critical examination, by health professionals, of their own 'culture of health', and that acknowledges the knowledge and wisdom of women. I suggest that a pedagogical relationship between health care professionals and women can be a site for the development of transformative knowledge and the empowerment of women. PMID- 9731158 TI - Incorporating health and behavioral consequences of child abuse in prevention programs targeting female adolescents. AB - A study examining the health and behavioral consequences of child abuse was conducted among 263 parenting and 257 never-pregnant teens attending a reproductive health clinic. Both groups of teens identified the following major consequences: suicide, prostitution, school drop-out, crime and substance abuse. However, only parenting teens expressed interest in prevention programs that would address these consequences. Traditional child abuse prevention programs are focused on parenting issues and rarely address health and behavioral consequences of abuse. These health and behavioral consequences of abuse may make adolescents vulnerable to abuse their own children as well as interfere with their psychosocial development. Therefore, the authors recommend integrating health and behavioral issues into child abuse prevention programs. PMID- 9731159 TI - Girls in the 90s: a gender-based model for eating disorder prevention. AB - Girls in the 90s is an eating disorder prevention program for pre adolescent/adolescent girls which operationalizes contemporary theories of female development. Set right at the beginning of the continuum of food and weight preoccupation, the program teaches girls to recognize when they feel fat and encourages them to tell the stories that lie underneath. The program validates girls' experiences and feelings, reframes them in terms of female development and provides girls with an understanding of the societal pressures that they face. The flexibility of the program enables it to be used by women with different professional orientations, with different age groups and to address different health and social risks that girls face. The skills can be adapted for use in group and individual counselling situations and can be integrated into existing resources and practice. PMID- 9731160 TI - The concerns of lesbians seeking counseling: a review of the literature. AB - Whilst lesbians are no different in terms of psychological adjustment than heterosexuals, they do seek counseling for a variety of reasons and some of these are unique to their experiences as lesbians. Some of these reasons are related to difficulties in coming out, particularly in young lesbians, and some are to do with relationship problems. In most cases where lesbians seek counseling it is difficult to establish a rapport and therapeutic milieu without disclosure of sexual orientation even if this does not directly relate to the presenting problems. However, empirical evidence suggests that lesbians have difficulty in finding counselors who are not homophobic or heterosexist and who are sensitive to their mental health concerns. The debate about whether or not specialist counseling services should be developed or mainstream services improved will be discussed. PMID- 9731161 TI - Factors associated with breast self-examination behaviour among Chinese women in Hong Kong. AB - The purpose of this study was to examine factors predicting the practice of breast self-examination behaviour among Chinese women in Hong Kong using a cross sectional survey research design. The Health Belief Model was used as the theoretical framework for the study to examine differences between breast self examination practicers and non-practicers and among breast self-examination frequency groups. Data were collected from a convenience sample of 124 women using self-administrated questionnaires. Less than half of the sample practised breast self-examination and only 16% of the practicers performing breast self examination monthly. There were differences between practicers vs. non-practicers and among breast self-examination frequency groups and some support of the predictive power of the Health Belief Model was found. Logistic regression showed that practicers perceived health as important, having fewer barriers and higher susceptibility to breast cancer. Discriminant function analysis revealed that barriers and children status were strong predictors of frequency of breast self examination practice explaining 21.1% of the total variance in breast self examination practice. PMID- 9731162 TI - The information needs of well, longer-term survivors of breast cancer. AB - Nine focus groups for well, longer-term survivors of breast cancer were held in Ontario, Canada. Prevalent themes identified through analysis of focus group transcripts fell into two broad categories, one reflecting the context within which women seek information and the other reflecting the content of information desired and sought. Themes related to context included: the ongoing impact on women of their initial disease experience and continued uncertainty about possible recurrence; womens' lack of information and understanding about processes involved in developing medical knowledge; prevailing mistrust about the impact of cost curtailment policies; and, concerns related to how professional communication can aid or hinder the goal of obtaining information. Themes related to content issues included: follow-up protocols, tamoxifen, detecting signs of possible recurrence, prevention for daughters, neglected side effects of treatment, insurance, lifestyle, and unconventional therapies. PMID- 9731163 TI - Woman-to-woman support: lessons from an Australian case story. AB - The purpose of this article is to demonstrate the use of storytelling to generate critical thinking about women's mental and emotional health care. The case story at the heart of the storytelling process describes a real, conflict situation in a remote, Australian women's health center. The themes generated lead to critical analysis of the concepts of care, independence, self-responsibility, ideology and woman-space, as they pertain to services designed to promote women's mental and emotional well-being. The conflict in the story arose between a lay worker providing woman-to-woman support and a professional counselor. Woman-to-woman support is a strategy developed in the women's health movement. It refers to the provision of time and space in women's health centers for women to work on their own needs with the support of women's health workers. Based on feminist principles, the aim is to empower women with knowledge, resources and skills. Woman-to-woman support builds on the care that women have traditionally provided for each other and is particularly useful for women living in regions where only limited, professional counseling services may be available. PMID- 9731164 TI - A decision aid for women considering hormone therapy after menopause: decision support framework and evaluation. AB - Although postmenopausal women are advised to consider their values when deliberating about potential benefits and risks of hormone therapy (HRT), feasible, effective methods of decision support in primary care have yet to be established. Using an explicit decision support framework, we developed a self administered HRT decision aid and evaluated it in a before/after study of 94 women from six family practices. An audiotape guided women through an illustrated booklet including: detailed information about HRT benefits and risks tailored to a woman's clinical risk, and a values clarification exercise to promote informed decision making consistent with personal values. After using the decision aid participants: had better general knowledge and more realistic personal expectations of HRT benefits and risks; and, felt more certain, informed, clear about values, and supported in decision making. Women's values elicited in the clarification exercise were 84% accurate in discriminating between decisions. Women with polarized preferences at baseline did not change their minds, but were better informed. Changes in preferences occurred in the uncertain group, with equal numbers accepting or declining HRT. Most participants found the decision aid comprehensible, acceptable in length and pace, and balanced. Decision aids are useful in preparing women for decision making about this complex, personal issue. PMID- 9731165 TI - Reassuring the woman facing menopause: strategies and resources. AB - Reassurance and reliable information are the prime requirements of women in premenopause, perimenopause and postmenopause. Women in premenopause want to know what to expect; women in perimenopause want to know that their experiences fall within the range of 'normal'; women in peri- and postmenopause need help in making sensible decisions about the use of hormone therapy. Based on the concerns expressed in over 7000 letters and other messages sent to the international newsletter, A Friend Indeed, this article discusses the concerns of North American women facing menopause in the last decade of the 20th century. PMID- 9731166 TI - Hormone replacement therapy: determinants of women's decisions. AB - Hormone replacement therapy: determinants of women's decisions. The purpose of this qualitative study was to explore the decision-making process used by menopausal women initiating or remaining on hormone replacement therapy (HRT), stopping HRT, or never starting HRT. Eight focus groups, composed of women reflecting these categories, were conducted. Four major themes or spheres of influence emerged as important in the women's decision-making process: the woman's internal influence--the interface between her perceptions and feelings including the symptoms of menopause, the benefits realized by HRT usage, and the experiences of negative side effects; interpersonal relationships, including the patient-physician relationship, family, friends and information networks; external influences, such as ageism and sexism; and consequences resulting from whichever treatment decision was chosen. A new concept was elucidated called "weighted influence" to underscore the dynamic interplay among the spheres. As information about HRT continues to grow and change, an understanding and application of these spheres of influence can assist physicians in engaging in a dialogue with their patients that allows individual evaluation and application of this new information. PMID- 9731167 TI - Building a feminist theoretical framework for screening of wife-battering: key issues to be addressed. AB - The purpose of this paper is to identify key questions which must be addressed in developing a woman-centred, that is feminist, theoretical framework for screening for wife battering in health settings. The health sector has not had a positive history in addressing women's health holistically or in terms of the social context of women's lives. This is notable in relation to the issue of wife battering, where attention is recent but growing and responses have been varied. On the other hand, violence against women is one of the greatest threats to their health and, therefore, it can be argued that the health sector has a major role to play in ending this violence. With the recent efforts within the health sector, a great deal has been accomplished; we have some consensus on what is an appropriate protocol for and the roles of health care providers. We are also making progress in developing training programs. Several things are still lacking an understanding of the systemic barriers to promotion of and participation in screening; sufficient program evaluations; sufficient attention to issues of diversity among women; and, a theoretical framework for practice which links these together. PMID- 9731168 TI - Patient education: new approaches and classical themes. PMID- 9731169 TI - The patient from receiver of information to informed decision-maker. AB - In this article the change of paradigm currently taking place within the field of patient education is indicated. In this change of paradigm the patient is increasingly being seen as responsible for his own health and as someone who makes independent choices in this respect. This has consequences for the role the patient is given in decisions on individuals care. The consequences of current developments within the Dutch society and in medical techniques as these relate to the need for patient education are enlightened. Some of the main topics for patient education are mentioned and the effects of patient education as established through meta-analyses are summarised. A planning model for a systematic development of patient education interventions is recommended. Relevant topics for research on patient education in the Netherlands are indicated. PMID- 9731171 TI - Cystic fibrosis through a female perspective: psychosocial issues and information concerning puberty and motherhood. AB - The purpose of the study was to investigate psychosocial issues concerning puberty and motherhood among CF adult females, to see how they had obtained and conceived information on these matters and how they would like information to be given. Fourteen adult CF females were interviewed. The majority of the women felt socially accepted and did not remember being ashamed over their delayed puberty. Thirteen of the women had been or were living in stable sexual relationships. However, the study revealed problems with destructive behaviour during puberty due to thoughts about premature death, secret worries over delayed puberty, poorly received information about puberty and fertility, avoidance of close relationships with the opposite sex during adolescence and concerns about being a mother with a chronic illness. Information about puberty and fertility should be given individually and in small discussion groups with teenage girls combined with thorough medical and psychological guidance concerning motherhood. PMID- 9731170 TI - Predictors of fecal occult blood screening among older socioeconomically disadvantaged Americans: a replication study. AB - Socioeconomically disadvantaged elderly are less likely to participate in fecal occult blood testing (FOBT). A quasi-experimental design was used in this operational replication study to determine predictors at baseline of subsequent participation in FOBT. Sixty-five percent of the 211 participants in the replication study participated in FOBT, and 47% of the 171 participants in the original study participated in FOBT. Predictors for FOBT in the replication study were male gender, age of 65-75 years old, ability to go places without assistance, history of having had a digital rectal examination and FOBT. This replication study supports targeting socioeconomically disadvantaged populations for FOBT as well as females, persons 85 years and older, persons who need assistance in travel, and person who have not had FOBT before. The results show that socioeconomically disadvantaged persons will participate in FOBT when effective educational interventions that include adaptation for aging changes are used. PMID- 9731172 TI - Randomized trial of a patient-centered hospital unit. AB - Patient-centered hospital units have grown out of the national trend to greater consumerism, but few of these units have been evaluated rigorously. We used a randomized controlled trial to compare patient outcomes on the Planetree Model Hospital Unit with other medical-surgical units in the hospital. Planetree patients were significantly more satisfied than controls with their hospital stay, the unit's environment and nursing care, but did not differ in ratings of physician care. Planetree patients reported more involvement in their care while hospitalized and higher satisfaction with the education they received. There were few differences between Planetree and controls in health behaviors. While Planetree patients reported better mental health status and role functioning after discharge, their health status was similar to controls after 3 to 6 months. There were no differences in length of stay and charges for the index hospitalization, readmissions or outpatient care during the following year. PMID- 9731173 TI - Family stress management following acute myocardial infarction: an educational and skills training intervention program. AB - Although the experience of acute myocardial infarction (AMI) is a family affair, little has been available to guide stress and distress reduction efforts focusing on all members of the family compared to the somewhat larger literature addressing stress management interventions with cardiac patients. This article provides a conceptual background for a specific behavioral therapy approach to family stress management in dealing with the sequelae of AMI for all family members with the goal of reducing morbidity for all family members as they cope with ongoing survivorship issues. The family intervention program is described and its pilot implementation discussed. Evaluation of the pilot suggests that an individually tailored focus for that subset of families at higher risk for elevated persistent distress may be the most cost-effective use of such a family intervention program. PMID- 9731174 TI - Specification of social support behaviours and network dimensions along the HIV continuum for gay men. AB - This study aimed to investigate social supports that HIV-infected persons find helpful and unhelpful, and the size and composition of networks along the disease continuum. Ninety six HIV-infected and 33 seronegative gay men were interviewed. The HIV continuum was represented by seronegative, HIV asymptomatic and symptomatic groups. Emotional and physical support were the most frequently identified helpful supports. Symptomatic persons identified physical support as helpful more often than asymptomatic persons. Availability, acceptance and nurturing were the most frequently identified helpful emotional support behaviours, while domestic support was the most frequently identified physical support behaviour. The most frequently mentioned unhelpful support was overprotectiveness. Overall, HIV-infected people had adequate social networks. Composition of the networks of HIV-infected persons differed from that of seronegative participants in that the former had markedly more professional and family persons and fewer friends in their network. HIV education and counseling interventions should provide emotional support, facilitate physical support for symptomatic persons, offer support that matches specific needs, include significant others, incorporate peer-help and be gay-sensitive. PMID- 9731175 TI - Personal resources, motives and patient education leading to changes in cardiovascular risk factors. AB - The purpose of this study was to assess the importance of patients' abilities and motivation to reduce their excess morbidity by benefiting from patient education designed to effect such change. The course consisted of a 4 week full-time program and 4 day refresher course 1 year later. The 295 consecutive patients (60%) who returned for the refresher had substantially reduced their overrisk for stroke or coronary heart disease. Personal ability to accomplish the desired change, such as standard of education, was found to be unimportant, whereas motives like having other people dependent on one were deemed important. One reason why resources such as education were found not to be important could be that the course was lengthy, requiring nearly 5 weeks, and practically oriented. PMID- 9731176 TI - The prognostic importance of patient pre-operative expectations of surgery for lumbar spinal stenosis. AB - The influence of psychosocial variables in the outcome of surgery for lumbar stenosis (LSS) has not been evaluated. We studied 257 patients with LSS pre operatively and at 6 months to: (a) relate patient expectations of surgery to baseline function and pain; and (b) determine how patient expectations and pre operative function interact to predict post-operative outcomes. RESULTS: On average, patients experienced substantial pain relief, improved function and satisfaction. Patients with many pre-operative expectations, particularly patients with low baseline function, reported more improvement in post-operative function than patients with few expectations. More ambitious expectations for physical function were also associated with improved function and satisfaction at 6 months. Conversely, having more numerous pain relief expectations was associated with more pain and less satisfaction with pain relief. CONCLUSION: Patient expectations influence recovery from surgery at 6 months. To improve outcomes and satisfaction, clinicians should discuss expectations with patients pre-operatively. PMID- 9731177 TI - Do the bacterial flagellar motor and ATP synthase operate as water turbines? AB - Despite much progress in the study of the rotary motors ATP synthase (F0F1) and the bacterial flagellar motor, we still cannot answer the most basic and simple question, how the random thermal movement of H+ (or Na+) ions down a pH (or Na+) gradient spins the rotors. I suggest consideration of the possibility that the motors operate like water turbines, i.e., rotation is the outcome of water mini jets impinging tangentially on the rotors. The vectorial jets are formed when the cations lose part or all of their hydration water upon interacting with a properly positioned site on a protein component which is part of the rotor or is located on a peripheral protein. PMID- 9731178 TI - On the coordination and oxidation states of the active-site copper ion in prokaryotic Cu,Zn superoxide dismutases. AB - The active-site copper ion of the prokaryotic Cu,Zn superoxide dismutase from P. leiognathi is found to undergo reversible reduction upon irradiation of the protein solution with a high-intensity X-ray beam from a third-generation synchrotron source. The same phenomenon is observed for the enzyme crystals, whose diffraction pattern has been obtained from synchrotron sources. In this case the active-site copper-ligand coordination bond lengths and in particular the Cu-NE2(His61) distance are consistent with a copper ion in the reduced state. These results are in line with previous studies on the eukaryotic Cu,Zn superoxide dismutases and suggest the conservation of an identical catalytic mechanism in both the prokaryotic and eukaryotic enzymes. PMID- 9731179 TI - Mechanism of action of a new component of the Ca(2+)-dependent proteolytic system in rat brain: the calpain activator. AB - Rat brain contains a calpain activator specific for the mu-form of the proteinase. We now report that this protein factor binds to the catalytic 80 kDa calpain subunit, promoting the dissociation of the heterodimer structure of the proteinase. The successive steps of the activation process, namely the two autoproteolytic steps producing the 78 kDa and the 75 kDa calpain forms, result in a 100 times faster rate. The activator competes with calpastatin and associates with the inner surface of plasma membranes. Based on its properties, the calpain activator can be visualised as the molecule indicating the sites for calpain activation at which the proteinase can also elude the negative control exerted by calpastatin. PMID- 9731180 TI - Bacterial overexpression, purification, and reconstitution of the carnitine/acylcarnitine carrier from rat liver mitochondria. AB - The carnitine/acylcarnitine carrier from rat liver mitochondria was overexpressed in Escherichia coli. The expressed protein, recovered as inclusion bodies, was solubilized with sarkosyl and purified by Sephadex G-200 and celite chromatography. A yield of 15 mg of purified transport protein per liter of cell culture was obtained. Upon reconstitution into liposomes, the purified carrier catalyzed a [3H]carnitine/carnitine exchange inhibited by maleimides, mercurials, and sulfobetaines. Carnitine esters of various lengths were also transported. The Km for carnitine uptake was 0.47 +/- 0.11 mM, the Vmax of the exchange was 0.78 +/- 0.24 mmol/min per gram of protein, and the Ki for octanoylcarnitine was 13.5 +/- 4.3 microM. The transport properties of the recombinant carrier were virtually identical to those of the native transporter. These studies represent the first overexpression of the functionally active mitochondrial carnitine/acylcarnitine carrier, thus enabling structure/function analysis of this protein by site-directed mutagenesis. PMID- 9731182 TI - Oscillations of membrane potential across a polypeptide membrane, induced by an electrical current. AB - Oscillation of membrane potential across a tri-block copolypeptide membrane composed of (Glu)x-(Leu)y-(Glu)x (x = 0.18 and y = 0.64) was observed under an electrical current, when the membrane was placed between equimolar aqueous salt solutions. The amplitude of the oscillation was influenced by the type of cation and anion in the external salt solution, and the amplitude was in the sequence: K+ > Na+ > Cs+ > Ca2+ and Cl- > Br-. The frequencies of the oscillations were in the range 0.1 to 5 Hz, and were also slightly influenced by the type of cation and anion. PMID- 9731181 TI - ZK1, a novel Kruppel-type zinc finger gene, is induced following exposure to ionizing radiation and enhances apoptotic cell death on hematopoietic cells. AB - To identify and early response factor by exposure to ionizing radiation, we cloned the cDNA of a novel Kruppel-type zinc finger (ZNF) gene, ZK1, from a cDNA library constructed from the human leukemia cell line, CMK86, using degenerate primers. This cDNA encoded a protein of 671 amino acids with an A box of Kruppel associated box (KRAB) domain at the N-terminus, followed by 15 ZNF motifs. The expression level of the ZK1 mRNA in human leukemia cells lines, CMK86 and U937, was increased after exposure to ionizing radiation. Furthermore, murine myeloid precursor 32D cells that were stably transfected with ZK1 cDNA had higher sensitivity to ionizing radiation than the 32D parent cells. These data suggest that the ZK1 gene is one of early response genes by exposure to ionizing radiation, and may have some functions on radiation-induced apoptotic cell death on hematopoietic cells. PMID- 9731183 TI - Evidence suggesting the involvement of hematopoietic stem cells in the pathogenesis of IgA nephropathy. AB - To investigate the role of hematopoietic stem cells in the pathogenesis of IgA nephropathy, T-cell-depleted bone marrow cells for IgA nephropathy-prone ddY mice were transplanted into C57BL/6j (B6) mice pretreated with cyclophosphamide. In the 12th week after bone marrow transplantation, transplanted bone marrow cells had successfully regenerated. In B6 recipients of T-cell-depleted allogeneic bone marrow cells for ddY mice ([ddy-->B6]), mesangial IgA and C3 deposits were significantly more intense than those in B6 mice receiving syngeneic bone marrow cells of B6 mice ([B6-->B6]). The serum IgA level in [ddY-->B6] mice was higher than that in [B6-->B6] mice. Molecular profile analysis of serum IgA revealed that the serum concentration of macromolecular IgA was increased in [ddY-->B6], but not in [B6-->B6] mice. These data suggest that disorders programmed at the level of BMCs are involved in the pathogenesis of IgA nephropathy by increasing circulating levels of macromolecular IgA. PMID- 9731185 TI - A monoclonal antibody that recognizes a common carbohydrate epitope shared by various glycoproteins in human secretions. AB - A monoclonal antibody (mAb; a mouse IgM referred to as 1CF11) recognizing various human glycoproteins was obtained. While the immunoreaction of glycoproteins from human secretions including milk, saliva, and bronchus was demonstrated as a typical dose-responded S-shaped reaction curve on ELISA, no reaction was detected with milks and sera of animal origin as well as human serum. In the constituting polypeptides of the human milk secretory IgA molecule, only the secretory component was recognized by this mAb. Among various chemical treatments of the purified human milk lactoferrin (Lf), only either periodate or mild alkaline treatment abolished the immunoreactivity of the glycoprotein. A recombinant human Lf was not immunoreactive. Finally, the immunoreactive fragments were isolated from human milk Lf, which remained reactive with PAS reagent while lacking the previously reported N-glycans. These results strongly suggest that the mAb 1CF11 recognizes a new glycan O-glycosidically linked to glycoproteins in human secretions. PMID- 9731184 TI - Sildenafil inhibits phosphodiesterase type 5 in human clitoral corpus cavernosum smooth muscle. AB - Phosphodiesterases play an important physiological role by regulating the intracellular levels of cyclic nucleotides. In this study, we investigated the kinetic parameters of inhibition of phosphodiesterase (PDE) type 5 (EC 3.1.4.35, 3',5'-cyclic GMP phosphodiesterase) by a novel, high-affinity, selective PDE type 5 inhibitor, sildenafil, in intact cells and in soluble extracts of human clitoral corpus cavernosum smooth muscle cells. Sildenafil inhibited cGMP hydrolysis in the crude extract (Ki = 7.2 +/- 2.7) and in partially purified preparations (Ki = 9 nM) in a competitive manner, as determined by Dixon plots. Sildenafil was a more effective PDE type 5 inhibitor than zaprinast (Ki = 400.0 +/- 76.4 nM, crude extracts; 250 nM, partially purified). Stimulation of intracellular cGMP synthesis by the nitric oxide donor sodium nitroprusside resulted in a 3.3- and 2.9-fold increase in cGMP concentration in the presence of sildenafil or zaprinast, respectively, compared to sodium nitroprusside treatment alone in intact cells at physiological temperatures. These observations suggest that human clitoral corpus cavernosum smooth muscle tone may be regulated by the synthesis and release of nitric oxide and that this pathway is dependent on PDE type 5 activity. PMID- 9731186 TI - Molt-inhibiting hormone mRNA levels and ecdysteroid titer during a molt cycle of the blue crab, Callinectes sapidus. AB - Synthesis of ecdysteroid molting hormones by crustacean Y-organs is believed to be regulated by a neuropeptide, molt-inhibiting hormone (MIH), produced in eyestalk neural ganglia. In the present study, steady-state MIH mRNA and hemolymph ecdysteroid levels were determined by Northern blot and radioimmunoassay, respectively, during the molt cycle of the blue crab, Callinectes sapidus. The level of MIH mRNA dropped steadily during premolt (D1 D4), reaching a minimum in D3/D4, then increased by 10-fold in postmolt (A/B) and remained elevated during intermolt (C4). These stage-specific changes in MIH mRNA levels were accompanied by significant fluctuations in the hemolymph ecdysteroid titer. The ecdysteroid titer increased steadily to a peak of 377.0 ng/ml in D3 of premolt, then dropped to 120.0 ng/ml in D4 (just prior to molting), and was low during postmolt (A/B, 4.4 ng/ml) and intermolt (C4, 3.3 ng/ml). The results represent the first report of developmental changes in MIH gene expression and are generally consistent with the hypothesis that MIH negatively regulates ecdysteroid synthesis in crustaceans. PMID- 9731187 TI - Bcl-2 is located predominantly in the inner membrane and crista of mitochondria in rat liver. AB - Bcl-2 is now recognized as a potent inhibitor of apoptotic cell death. It has been reported that Bcl-2 is located in the mitochondria, endoplasmic reticulum, and nuclear membrane in some cell lines, and it is not expressed in normal human and rat liver. An earlier study showed that Bcl-2 is an inner mitochondrial membrane protein. On the contrary, the following investigations using immunoelectron microscopy demonstrated that Bcl-2 resides predominantly in the mitochondrial outer membrane. In this study, using a cryo-sectioning immunogold labeling technique and immunoblotting, we carefully determined the subcellular localization of Bcl-2. Here we report that Bcl-2 is expressed in normal rat liver, and it is located predominantly in the inner membrane and crista rather than in the outer membrane of mitochondria. PMID- 9731188 TI - Differences in cytotoxicity of native and engineered RIPs can be used to assess their ability to reach the cytoplasm. AB - Ricin is a heterodimeric cytotoxin composed of RTB, a galactose binding lectin, and RTA, an enzymatic N-glycosidase. The toxin is endocytosed, and after intracellular routing, RTA is translocated to the cytoplasm where it inactivates ribosomes resulting in a loss of host cell protein synthesis and cell death. We show for the first time that the cytotoxicity against cultured T cells by several RTA mutants is directly proportional to the enzyme activity of RTA, suggesting this is a reliable system to measure translocation effects. Large discrepancies between cytotoxicity and enzyme action for a given pair of toxins are therefore attributable to differences in cell binding, uptake, or membrane translocation. Fluid phase uptake and cytotoxicity of isolated RTA are essentially identical to that of the single chain toxin PAP. This important finding suggests that RTA, and the A chain of class 2 RIPs in general, has not evolved special translocation signals to complement the increased target cell binding facilitated by RTB. Experiments with the lectin RCA and with ebulin suggest those toxins have diminished cytotoxicity probably mediated by comparative deficiencies in B chain binding. Addition of a KDEL sequence to RTA increases fluid phase uptake, consistent with the notion that transport to the ER is important for cytotoxicity. Fusion of MBP or GST to the amino terminus of RTA has little effect on enzyme action or cytotoxicity. This result is not altered by protease inhibitors, suggesting the fusion proteins are probably not cleaved prior to translocation of the toxic A chain and implying that the toxins can carry large passenger proteins into the cytoplasm, an observation with interesting potential for analytical and therapeutic chemistry. PMID- 9731189 TI - Colocalization of emerin and lamins in interphase nuclei and changes during mitosis. AB - Emerin is a nuclear membrane protein which is affected by mutation in X-linked Emery-Dreifuss muscular dystrophy. We have previously suggested that emerin is a member of a family of type II integral membrane proteins which associate with the nuclear lamina and which include lamina-associated proteins and the lamin B receptor. We now show that emerin in COS cells is not restricted to the nuclear rim but is also found at intranuclear sites, where it colocalizes with nuclear lamins B1, B2 and A/C. During mitosis, emerin is dispersed throughout the cell and then participates in the reconstitution of membranes around the daughter nuclei. Although emerin and lamins do not remain colocalized during mitosis, they all show some association with the midbody of the mitotic spindle. PMID- 9731190 TI - A splice junction mutation in the alpha(M) gene of phosphorylase kinase in a patient with myopathy. AB - In a 28-year-old man with myopathy and phosphorylase kinase (PhK) deficiency, we found a G-to-C substitution at the 5' end of an intron in the muscle-specific alpha-subunit gene. The mutation destroys the high-consensus GT sequences at the 5' splice junction of the intron, which causes skipping of the preceding exon. This is the second molecular genetic defect identified in the myopathic variant of PhK deficiency. PMID- 9731191 TI - Altered posttranslational modifications of collagen in keloid. AB - Keloid is a tissue with an excessive accumulation of collagen. In this study, we have partially characterized post-translational modifications of type I collagen in human keloid in order to pursue their potential involvement in this pathology. The levels of lysyl hydroxylation of the helical portions of alpha 1 and alpha 2 chains of type I collagen in keloid were significantly higher than those of normal, while the levels of prolyl hydroxylation were identical between these two groups. The contents of the major reducible cross-links in dermal collagen, dehydro-hydroxylysinonorleucine and dehydro-histidinohydroxymero-desmosine, were both significantly higher in keloids (up to sixfold) than those of normal. In addition, significant amounts of hydroxylysine-aldehyde derived cross-links that are characteristic of skeletal tissue collagens, dehydro dihydroxylysinonorleucine (about 0.3 mole/mole of collagen) and pyridinoline (about 0.1 mole/mole of collagen), were found in keloids. These results indicate that keloid-forming cells are phenotypically different from those in normal dermis and that the collagen produced is highly cross-linked. The increased cross linking provides the fibrils with more stability that may result in an accumulation of collagen. PMID- 9731192 TI - N-terminal myosin-binding fragment of talin. AB - Talin, an actin-binding protein from smooth muscle, is shown to bind to myosin in such a way that it stimulates the ATPase activity of myosin irrespective of the phosphorylation state of myosin. The binding site is shown to be localized at the N-terminal, 47 KDa fragment. The position of the actin-binding site at the C terminal suggests that talin may work as a crosslinker between myosin and actin. PMID- 9731193 TI - Sampling the genomic pool of protein tyrosine kinase genes using the polymerase chain reaction with genomic DNA. AB - The polymerase chain reaction (PCR), with cDNA as template, has been widely used to identify members of protein families from many species. A major limitation of using cDNA in PCR is that detection of a family member is dependent on temporal and spatial patterns of gene expression. To circumvent this restriction, and in order to develop a technique that is broadly applicable we have tested the use of genomic DNA as PCR template to identify members of protein families in an expression-independent manner. This test involved amplification of DNA encoding protein tyrosine kinase (PTK) genes from the genomes of three animal species that are well known development models; namely, the mouse Mus musculus, the fruit fly Drosophila melanogaster, and the nematode worm Caenorhabditis elegans. Ten PTK genes were identified from the mouse, 13 from the fruit fly, and 13 from the nematode worm. Among these kinases were 13 members of the PTK family that had not been reported previously. Selected PTKs from this screen were shown to be expressed during development, demonstrating that the amplified fragments did not arise from pseudogenes. This approach will be useful for the identification of many novel members of gene families in organisms of agricultural, medical, developmental and evolutionary significance and for analysis of gene families from any species, or biological sample whose habitat precludes the isolation of mRNA. Furthermore, as a tool to hasten the discovery of members of gene families that are of particular interest, this method offers an opportunity to sample the genome for new members irrespective of their expression pattern. PMID- 9731194 TI - A possible new role for vitamin D-binding protein in osteoclast control: inhibition of extracellular Ca2+ sensing at low physiological concentrations. AB - Upon removal of its sialic acid or galactose residue, vitamin D-binding protein (DBP) becomes a potent macrophage-activating factor, DBP-MAF. Here we document a new function of DBP-MAF and its parent molecule, DBP, in osteoclast control. We show that all DBPs potently inhibit extracellular Ca2+ (cation) sensing at low nanomolar concentrations with the following rank order of potency: native DBP = sialidase-treated DBP > beta-galactosidase-treated DBP. This attenuation remains unaffected despite co-incubation either with the native DBP ligand, 1,25 dihydroxyvitamin D3, or with an asialoglycoprotein receptor modulator, asialoorosomucoid. Taken together, the results suggest that circulating DBP may play a role in the systemic control of osteoclastic bone resorption, a hitherto unrecognized action of the protein. PMID- 9731195 TI - Janus kinase 2 (Jak2) must be catalytically active to associate with the AT1 receptor in response to angiotensin II. AB - Angiotensin II evokes a variety of biological responses by binding to a seven transmembrane cell surface receptor termed AT1. Ligand binding to the AT1 receptor induces the physical association and activation of the intracellular kinase Jak2. To elucidate the mechanism of this association, COS-7 cells were co transfected with the AT1 receptor and either wild type Jak2 or a catalytically inactive Jak2. AT1 receptor-Jak2 association was assessed in vitro by a GST-AT1 receptor fusion protein binding assay and in vivo by direct co immunoprecipitation of the receptor-Jak2 complex. Both studies showed that Jak2 must be catalytically active to form a complex with the AT1 receptor, and that complex formation is associated with Jak2 tyrosine phosphorylation. These results were confirmed using the Jak2 specific inhibitor AG-490. We also found that over expression of wild type Jak2 in COS-7 cells leads to in vivo complex formation of spontaneously autophosphorylated Jak2 with the AT1 receptor. No such complex formation was observed with a dominant negative Jak2. Thus, the physical association of Jak2 with the AT1 receptor is regulated by an angiotensin II mediated autophosphorylation event. PMID- 9731196 TI - Marked enhancement in the reductive dehalogenation of hexachloroethane by a Thr319Ala mutation of cytochrome P450 1A2. AB - Mutation of the conserved Thr319 residue to Ala of cytochrome P4501A2 (CYP1A2) increased the value of Vmax 9-fold for reductive dehalogenation of hexachloroethane in the reconstituted system under anaerobic conditions. The Thr319Ala mutation also increased the elimination over substitution product ratio by 5-fold. The addition of aliphatic alcohols increased by 22-fold the activity obtained with the wild type and varied the elimination over substitution product ratio. Increasing pH increased the ratio of elimination over substitution by primarily affecting the rate of elimination. PMID- 9731197 TI - Thioredoxin peroxidase (natural killer enhancing factor) regulation of activator protein-1 function in endothelial cells. AB - Thioredoxin peroxidase-1 (TxP-1), originally cloned as natural killer enhancing factor-B, belongs to a highly conserved antioxidant family. Tumor necrosis factor alpha (TNF) activates the expression of activator protein-1 (AP-1) responsive genes. We show here that over-expression of TxP-1 blocks TNF-induced AP-1 activation in endothelial ECV304 cells, which was demonstrated by three independent experimental protocols: electromobility shift assay with AP-1 oligonucleotide probe; reporter gene expression with AP-1 binding site, and interleukin-8 production, which is dependent on AP-1. These results are consistent with the role of TxP-1 as an antioxidant and the previous reports that TNF-induced reactive oxygen species were responsible for AP-1 activation. PMID- 9731198 TI - Cellular responses in mouse leukemia L1210 cells made resistant to deoxyadenosine. AB - Recent studies have implicated nucleotides in diverse and unexpected functions related to p53 levels, p53-dependent G0/G1 cell cycle arrest, and the role of dATP in the activation of the caspase-induced apoptosis. Using deoxyadenosine resistant L1210 cells (ED2 and Y8) that had ribonucleotide reductase that was not sensitive to inhibition by dATP and also exhibited other metabolic alterations, the properties of these cells with respect to the role(s) of nucleotides in these functions were explored. In the ED2 and Y8 cells that did not express p53 protein, the pools of UTP, CTP, ATP, and GTP were markedly decreased. The decreased cellular levels of UTP and CTP did not result in these cells being more sensitive to either PALA or acivicin. The ED2 and Y8 cells did not block in G0/G1 in response to PALA treatment even though the basal cellular concentrations of UTP and CTP were reduced 50 to 80%. While it has been shown that dATP in combination with cytochrome c is involved in the apoptotic pathway, the concentration of exogenous deoxyadenosine required to induce apoptosis in the parental L1210 cells was far in excess of the concentration required to inhibit cell growth. Deoxyadenosine did not cause an increase in apoptosis in the deoxyadenosine-resistant Y8 cells. These data suggest that the new roles ascribed to nucleotides may be specific for the particular cell type under very specific conditions. PMID- 9731199 TI - MKP-1 induced in rat brain after electroconvulsive shock is independent of regulation of 42- and 44-kDa MAPK activity. AB - Electroconvulsive shock (ECS) activates MAPKs in rat brain and also induces immediate early genes. We investigated whether ECS induces MKP-1, a specific MAPK phosphatase and an immediate early gene, for feedback regulation of MAPK activity. ECS induced MKP-1 in the cortex, but MAPK activity returned to its basal level before MKP-1 protein increased, within 10 min of ECS. MKP-1 protein amount peaked 1 hr after ECS. MKP-1 induced did not lower the basal level of MAPK activity or attenuate MAPK activation by second ECS. MAPK activation in cerebellum was very weak, but the MKP-1 induction was faster and more prominent than in the cortex. These results suggest that ECS induces MKP-1 in various rat brain regions, however, the induction may not be related to the activation of MAPK and the MKP-1 induced may be independent of the regulation of MAPK activity after ECS. PMID- 9731200 TI - Human dynamin-like protein interacts with the glycogen synthase kinase 3beta. AB - Members of the dynamin superfamily are implicated in vesicle trafficking. Using human glycogen synthase kinase 3 beta (Gsk-3 beta) as bait in the yeast two hybrid system, we identified a novel human dynamin-like protein IV (HdynIV). When the full-length cDNA of HdynIV was sequenced, it showed that HdynIV's carboxyl terminal lacks a proline-rich domain that can bind to Gsk-3 beta. By Northern blot analysis and isoform-specific PCR, we found that HdynIV is expressed ubiquitously in all human tissues examined. Two transcripts of 2.4 and 4.4 kb are shown to be more abundant in heart, brain, and skeletal muscle. Interestingly, the 2.4-kb transcript is expressed more distinctly in the fetal liver than in the adult liver, suggesting that this protein might play a role during development. In the present report, we have demonstrated that HdynIV interacts with the Gsk-3 beta through its carboxyl-terminal region, implying than HdynIV may also be involved in cell signaling. PMID- 9731201 TI - Thyroid hormone regulates the acetyl-CoA carboxylase PI promoter. AB - The acetyl-CoA carboxylase-alpha gene has two promoters, PI and PII. A variety of mRNA products result from this gene, depending on promoter usage and splicing events. We have investigated thyroid hormone regulation of acetyl-CoA carboxylase alpha gene expression, using the reverse-transcription polymerase chain reaction with PI- or PII-specific primers. RNA was extracted from a range of tissues taken from hypo-, eu-, or hyperthyroid rats. PII-generated products were found in all tissues examined at similar levels and were not affected by thyroid state. Products derived from PI were also widely found but with more variable levels of expression. PI mRNAs were reduced in hypo- and elevated in hyperthyroid livers. In brown adipose tissue, more PI products were found in hypothyroid animals. Thus, thyroid hormone regulates the activity of the acetyl-CoA carboxylase PI promoter to influence fatty acid synthesis in a tissue-specific manner. PMID- 9731202 TI - Hypoxia exerts cell-type-specific effects on expression of the class 3 aldehyde dehydrogenase gene. AB - The Class 3 aldehyde dehydrogenase gene (ALDH3) is expressed differentially in a tissue-specific manner, occurring constitutively in some tissues and in others as a result of xenobiotic induction via the Ah receptor/ARNT pathway. ARNT is also involved in regulating gene expression in response to hypoxia. It dimerizes with hypoxia-inducible factor 1 alpha (HIF-1 alpha) and enhances expression of hypoxia responsive genes. To determine if ARNT plays a role in regulating ALDH3 in response to low oxygen tension, we studied the effects of 1% oxygen and the hypoxia mimic cobalt chloride on constitutive and inducible ALDH3 expression in rat hepatoma cells and rat corneal epithelial cells. Hypoxia sharply down regulates constitutive ALDH3 expression in corneal epithelial cells. Likewise, aromatic hydrocarbon-induced ALDH3 expression in H4-II-EC3 cells is significantly reduced by hypoxia. In contrast, hypoxia has no effect on constitutive or aromatic hydrocarbon-inducible ALDH3 expression in HTC cells. Our data indicate that hypoxia exerts cell type-specific effects on both constitutive and induced ALDH3 expression. PMID- 9731203 TI - Characterisation of porcine peroxisome proliferator-activated receptors gamma 1 and gamma 2: detection of breed and age differences in gene expression. AB - Two isoforms of peroxisome proliferator-activated receptor gamma (PPAR gamma) cDNAs, gamma 1 and gamma 2, have been isolated and characterised in swine. The relative expression of the two transcripts was studied by northern blot analysis using total RNA isolated from several porcine tissues taken at three different ages (day 1, after 5 weeks and at 100 kg weight). Hybridisation were carried out with two different probes, one binding to both PPAR gamma transcripts and the other being PPAR gamma 2 specific. Strongest hybridisation signals with the PPAR gamma probe binding both variants were detected in adipose tissues and spleen at all three ages, whereas only faint or no signals were detected in other tissues. The tissue distribution pattern of PPAR gamma 1 and gamma 2 suggests a modulation of tissue distribution for the two transcripts and obvious age and breed differences in gene expression in swine. PMID- 9731204 TI - Hydroxamate iron complex with phenoloxidase activity acting on lignin and chlorolignins. AB - The properties of a siderophore model, acetohydroxamic acid (AHA), of desferral were studied. The pH, ionic strength, and AHA/Fe(III) ratios for o-dianisidine oxidation were optimized. Phenoloxidase activity of hydroxamates/Fe(III) acting on o-dianisidine at pH 7.0 and pH 3.0 was observed. Under these conditions, AHA was able to reduce Fe(III) to Fe(II) followed by ferrozine complexation. AHA/Fe(III) complex degraded lignin and chlorolignins from kraft effluent E1 65% and 85%, respectively, after 24 h. PMID- 9731205 TI - Effects of RIalpha overexpression on cisplatin sensitivity in human ovarian carcinoma cells. AB - Our laboratory has found that Chinese hamster ovary (CHO) and mouse Y1 adrenocortical carcinoma PKA mutants with a defective R subunit, but not altered C subunits, exhibit increased resistance to cisplatin as well as other DNA damaging agents. The mechanism of resistance may be associated with increased recognition of the cisplatin-damaged DNA and protein binding to the DNA lesion, thus enhancing DNA repair in the RI alpha mutants. These data suggest that mutation of RI alpha may confer resistance to cisplatin by affecting DNA repair activity. In the present study, we overexpressed RI alpha in human ovarian carcinoma A2780 cells to demonstrate that RI alpha can modulate cellular sensitivity to cisplatin. Retroviral-infected A2780 cells overexpressing wild type RI alpha cDNA displayed a four- to eightfold greater sensitivity to cisplatin compared with parental cells. Overexpression of RI alpha in the CP70 cisplatin-resistant derivative of A2780 also increased the sensitivity of these cells to cisplatin. Therefore, enhanced expression of the RI alpha subunit of PKA sensitizes cells to the cytotoxic effects of this DNA-damaging agent. These data suggest that RI alpha may act directly, independent of the C subunit, to influence cellular sensitivity to cisplatin. Therefore, modulation of RI alpha expression or its functional status by pharmacological agents may potentially reverse cisplatin resistance in tumors. PMID- 9731206 TI - Association of Cdk2/cyclin E and NF-kappa B complexes at G1/S phase. AB - NF-kappa B/Rel family plays a pivotal role in a wide variety of cellular functions including growth, development, apoptosis and stress responses. Recent studies indicated that NF-kappa B is also involved in the cell cycle regulation, and high expression of c-Rel results in a cell cycle arrest at the G1/S-phase transition (Bash, J., Zong, W,-X., and Gelinas, C. (1997) Mol. Cell. Biol. 17, 6526-6536). Here we report the detection of Cdk2, a critical kinase responsible for the G1/S-phase transition, in immune complexes precipitated by the NF-kappa B antisera. Cdk2 and NF-kappa B association was detected by co-precipitation in the nuclear lysates of the G1/S-phase cells, and was found in cultured cell lines and in T cells purified from human peripheral blood. Using an affinity column containing the C-terminal peptide of human c-Rel, we isolated cyclin E, the regulatory subunit of the Cdk2 complex, as a c-Rel-binding protein. These findings support and provide physical basis for the involvement of NF-kappa B in the G1/S-phase cell cycle control, and suggest an important role played by the C terminal sequence of c-Rel. PMID- 9731207 TI - Combinational effects of vitamin D3 and retinoic acid (all trans and 9 cis) on proliferation, differentiation, and programmed cell death in two small cell lung carcinoma cell lines. AB - The effects of a combination of vitamin D3 [1,25(OH)2D3] and retinoic acid (RA) on proliferation, differentiation, and apoptosis of the human small cell lung carcinoma (SCLC) cell lines NCI-H82 and NCI-H209 were evaluated. Cell proliferation was inhibited by 1,25(OH)2D3 and RA alone. The combination of 1,25(OH)2D3 and the cis form of retinoic acid resulted in an additive decrease in cell proliferation and the induction of apoptosis in various concentrations. Moreover, 3H-thymidine incorporation was inhibited and the number of viable cells was decreased. The characteristics of the apoptotic cells were examined and confirmed by morphologic analysis, light and electron microscopy, and fluorescence detection. It was concluded that 1,25(OH)2D3 and RA exert additive effects on the inhibition of proliferation and the induction of apoptosis in both the NCI-H82 and the NCI-H209 SCLC cell lines. This finding has important implications for the use of retinoids and 1,25(OH)2D3 in cancer prevention and in the therapy of small cell lung carcinoma. PMID- 9731208 TI - An allosteric drug, o,o'-bismyristoyl thiamine disulfide, suppresses HIV-1 replication through prevention of nuclear translocation of both HIV-1 Tat and NF kappa B. AB - The efficacy of o,o'-bismyristoyl thiamine disulfide (BMT) was examined in detail against HIV-1 laboratory isolates (HTLV-IIIB, JRFL, and MN), primary isolates (KMT and KMO), and simian immunodeficiency virus (SIVmac251) in vitro. BMT inhibited the replication of HIV-1 in both laboratory and primary isolates in vitro. In addition, BMT exhibited antiviral activity against SIVmac251. Minimizing energy studies of BMT structure reveal that a trans-disulfide of thiamine (holo drug) disulfide (TDS, protodrug) is allosterically transited to the reactive twisted disulfide of BMT (allo drug) by o,o'-bismyristoyl esterification of TDS. BMT inhibits nuclear translocation of both HIV-1 transactivator (TAT) and the cellular transcriptional nuclear factor-KB (NF-kappa B), resulting in the suppression of HIV-1 replication. PMID- 9731210 TI - Nuclear import and nucleolar accumulation of the human ribosomal protein S7 depends on both a minimal nuclear localization sequence and an adjacent basic region. AB - In the course of the eukaryotic ribosomal biogenesis, both the nuclear import and export are involved. We have studied the nuclear and nucleolar localization of the human ribosomal protein S7. We examined the subcellular distribution of the S7:beta-galactosidase fusion protein in SAOS-2 cells. We have identified two evolutionarily conserved domains, both of which are necessary for S7 nuclear and nucleolar targeting: amino acids 98 to 109 and 115 to 118. Out of the S7 protein context, a fragment 98...118, containing these domains, is sufficient for nuclear transport and nucleolar accumulation. Interestingly, a tetrapeptide 115KRPR118, which can act as an independent nuclear localization signal (NLS), is not sufficient for exclusively nuclear accumulation of the S7 protein if the adjacent region 98...109 is deleted. In addition, site-directed mutagenesis revealed that critical residues for nuclear targeting in this tetrapeptide and in the full length S7 protein are different. While mutation of a Pro117 significantly impaired nuclear import of S7, similar substitution within the tetrapeptide-NLS had no effect on nuclear targeting. This suggests that to function perfectly, proper secondary structure of the S7 nuclear targeting domain is required. PMID- 9731209 TI - Point mutations (Thr240Arg and Gln311Stop) [correction of Thr240Arg and Ala311Stop] in the Parkin gene. AB - Autosomal recessive juvenile parkinsonism (AR-JP) is a distinct clinical and genetic entity characterized by selective degeneration of nigral neurons. Recently, the parkin gene responsible for AR-JP has been identified. To date, we found two different deletional mutations including single and multiple exonic deletions. In the present study, we identified two types of point mutations (Thr240Arg and Gln311Stop) involving exons 6 and 8 in the parkin gene of the AR JP patients from two Turkish families. This is the first report on point mutations for the parkin gene. Furthermore, the Thr240Arg mutation was located on a consensus sequence for the site of phosphorylation by casein kinase II. Identification of its mutation provides an important clue as to the role of the Parkin protein in degeneration of the substantia nigra in the brain of AR-JP patients. PMID- 9731212 TI - Exploration of the structural environment of the iron-sulfur cluster in putidaredoxin by nitrogen-15 NMR spectroscopy of selectively labeled cysteine residues. AB - Putidaredoxin is a di-iron protein whose paramagnetic region is not well characterized by 1H detected NMR. We have studied the structure of this region in greater detail by directly observed 15N NMR of oxidized and reduced putidaredoxin preparations in which the six cysteine residues are selectively labeled with 15N. A new method for preparation of a stable form of reduced putidaredoxin has been developed for use in NMR. The 15N NMR spectra of the oxidized and reduced forms are characteristically different, and we have measured and compared 15N chemical shifts, spin-lattice relaxation times (T1), and chemical shift/temperature dependences for both forms. Evidence for localized valencies of the iron atoms in the reduced form is presented. From the 15N T1 values of the oxidized form, reduced distances of the cysteine backbone 15N nuclei from the center of the Fe2S2 cluster have been calculated. These distances are consistent with those calculated from X-ray crystal structure data for five ferredoxins, and confirm the structural similarity of the Fe2S2 clusters in putidaredoxin and in these ferredoxins in the oxidized state. PMID- 9731211 TI - Generation of nitric oxide by a nitrite reductase activity of xanthine oxidase: a potential pathway for nitric oxide formation in the absence of nitric oxide synthase activity. AB - Nitric oxide (NO) synthesis is well-known to result from the oxidation of L arginine by a family of NO synthases (NOS). However, under hypoxic conditions this mechanism of NO synthesis may be impaired and NO is formed by a NOS independent mechanism. This study was designed to examine the reduction of nitrite to NO by xanthine oxidase (XO) under hypoxia, because the bacterial nitrate/nitrite reductases have structural similarity to XO. We found that both purified and tissue containing XO catalyze the reduction of nitrite to NO, as demonstrated using a chemiluminescent NO meter. This redox reaction requires NADH as an electron donor, and is oxygen independent. The inhibitory profiles suggest that reduction of nitrite takes place at the molybdenum center of XO whilst NADH is oxidized at the FAD center. Heparin binding of XO caused an increase in the catalysis of nitrite reduction. The XO-catalyzed generation of NO may be important in redistribution of blood flow to ischaemic tissue as a supplement to NOS, since both nitrite and NADH have been shown to be elevated in hypoxic tissue. PMID- 9731213 TI - Requirement of the serine-threonine kinase Akt for heat treatment-induced activation of p70 S6 kinase. AB - p70 S6 kinase plays an important role in growth factor-induced translational control and in cell cycle progression. Although the mechanism of p70 S6 kinase regulation is not fully understood, phosphorylation of serine and threonine residues of the enzyme is essential for its activation. The possible role of the serine-threonine kinase Akt in the activation of p70 S6 kinase induced by exposure of cells to heat has now been investigated. Overexpression of a mutant Akt1 (Akt-AA) in which the phosphorylation sites (Thr308 and Ser473) targeted by growth factors are replaced by alanine was shown to exert a dominant negative effect on Akt activation induced by platelet-derived growth factor (PDGF) or by heat treatment in CHO cells. Akt-AA also inhibited p70 S6 kinase activation induced by these stimuli. However, Akt-AA had no effect on the activation of p70 S6 kinase induced by 12-O-tetradecanoylphorbol 13-acetate, which did not stimulate Akt activity in these cells. These data suggest that Akt is required for heat treatment-induced activation of p70 S6 kinase. PMID- 9731214 TI - Overexpression of the Na(+)-Ca2+ exchanger leads to enhanced inotropic responsiveness to Na(+)-channel agonist without sarcoplasmic reticulum protein changes in transgenic mice. AB - The present study investigated the influence of over-expression of the cardiac Na(+)-Ca2+ exchanger on myocardial force of contraction and alterations in sarcoplasmic reticulum (SR) Ca(2+)-handling proteins in a transgenic mouse model. Inotropic effects of Na+ channel agonist BDF 9148 and isoprenaline were determined in isolated electrically driven atria. Protein levels of key SR Ca(2+) handling proteins were determined by Western blot analysis. Transgenic animals had no myocardial hypertrophy or failure. The positive inotropic effect of BDF 9148 was significantly more pronounced in myocardium for transgenic animals, whereas the inotropic response to isoprenaline was similar in both groups. Strong immunoreactivity of the transgene Na(+)-Ca2+ exchanger was detected in myocardium of transgenic animals. Protein levels of SR Ca(2+)-ATPase, phospholamban, and calsequestrin were unchanged. In conclusion, transgenic overexpression of the Na(+)-Ca2+ exchanger is accompanied by increased force development following Na+ channel agonist administration, even though Ca2+ proteins of the SR are unchanged. PMID- 9731215 TI - Identification of stathmin as a novel substrate for p38 delta. AB - p38 mitogen-activated protein kinases (MAPK) are a family of kinases that are activated by cellular stresses and inflammatory cytokines. Although there are many similarities shared by the isoforms of p38 (alpha, beta, gamma, and delta), p38 delta differs from the others in some respects such as inhibitor sensitivity and substrate specificity. Utilizing in a solution kinase assay, we identified a novel p38 delta substrate as stathmin. Stathmin is a cytoplasmic protein that was previously reported to be a substrate of several intracellular signaling kinases and has recently been linked to regulation of microtubule dynamics. p38 delta has significantly higher in vitro phosphorylating activity against stathmin than other p38 isoforms or related MAPKs. In transient expression studies, we found that in addition to different stimuli osmotic stress activates p38 delta to phosphorylate stathmin. The sites of phosphorylation were mapped to Ser-25 and Ser-38, both in vitro and in cells. PMID- 9731216 TI - Molecular cloning and characterization of the rat ovarian 20 alpha-hydroxysteroid dehydrogenase gene. AB - The rat 20 alpha-hydroxysteroid dehydrogenase (20 alpha-HSD) is an enzyme responsible for the catabolism of progesterone to the inactive 20 alpha hydroxprogesterone. We have previously shown that the expression of this enzyme is not regulated by post-translational modification, but at the level of transcription. In this study we have established that the 20 alpha-HSD gene contains nine exons and have isolated a 2.5 kb promoter region. The transcription start site was identified and a TATA box was found. 5' deletions of this promoter significantly decreased basal promoter activity. Treatment with forskolin led to a dose dependent inhibition of the 2.5kg-20 alpha-HSD-luciferase construct. Computer analysis identified one CRE, two Nur77 response elements, two putative AP1 sites and one progesterone response element half-site. In summary, we have identified and partially characterized the promoter region of the rat ovarian 20 alpha-HSD and demonstrated that the regulatory elements for 20 alpha-HSD are present within a 2.5 kb 5' flanking region of the gene. PMID- 9731217 TI - Hammerhead ribozymes targeted to the FBN1 mRNA can discriminate a single base mismatch between ribozyme and target. AB - Hammerhead ribozymes are catalytic RNA molecules that can act in trans, with ribozyme and substrate being two different oligoribonucleotides with regions of complementarity. Mutations in the gene for fibrillin-1 (FBN1) cause Marfan syndrome. The majority of mutations are single-base changes, many of which exert their effect via a dominant-negative mechanism. Previously we have shown that an antisense hammerhead ribozyme, targeted to the FBN1 mRNA can reduce deposition of fibrillin to the extracellular matrix of cultured fibroblasts, suggesting it may be possible to utilize ribozymes to down regulate the production of mutant protein and thus restore normal fibrillin function. This might be achieved by the mutation creating a ribozyme cleavage site that is not present in the normal allele, however this is likely to limit the number of mutations that could be targeted. Alternatively, it might be possible to target the mutant allele via the ribozyme binding arms. To determine the potential of ribozymes to preferentially target mutant FBN1 alleles via the latter approach, the effect of mismatches in helix I of a hammerhead ribozyme, on the cleavage of fibrillin (FBN1) mRNA was investigated. A single base mismatch significantly reduced ribozyme cleavage efficiency both in vitro and in vivo. The discrimination between fully-matched and mismatched ribozyme varied with the length of helix I, with the discrimination being more pronounced in ribozymes with a shorter helix. These data suggest that it should be possible to design hammerhead ribozymes that can discriminate between closely related (mutant and normal) target RNAs varying in as little as a single nucleotide, even if the mutation does not create a ribozyme cleavage site. PMID- 9731218 TI - Regulation of the Drosophila bHLH-PAS protein Sima by hypoxia: functional evidence for homology with mammalian HIF-1 alpha. AB - Hypoxia inducible factor-1 (HIF-1) is a heterodimeric complex of two basic-helix loop-helix proteins of the PAS family which is critical for oxygen-dependent expression of many mammalian genes. Regulation is mediated by the alpha subunit (HIF-1 alpha) and sequences from HIF-1 alpha can confer hypoxia-inducible activity on a Ga14 fusion protein. To analyse conservation of this system of gene regulation between Drosophila and mammalian cells we constructed Ga14 fusions with a series of Drosophila basic-helix-loop-helix PAS (bHLH-PAS) proteins and tested for hypoxia inducibility in transfected Hep3B cells. We found that Ga14 functions with Similar (Sima) but not other Drosophila bHLH-PAS proteins showed inducible activity following exposure to stimuli which classically activate mammalian HIF-1:hypoxia, cobaltous ions, and desferrioxamine. We also found that Sima protein accumulated in Drosophila SL2 cells following hypoxia. Together these findings indicate the existence of functional homologies between Sima and HIF-1 alpha, and that conservation is such as to enable Sima to interact with the hypoxia signal transduction system in mammalian cells. PMID- 9731219 TI - Cholesterol oxides induce programmed cell death in microglial cells. AB - N9 microglial cells were used as a model to examine the effect of cholesterol oxides on central nervous system microglia. Results indicated that 25-OH cholesterol was the most cytotoxic agent among the cholesterol oxides tested. During the process of cell death, this agent caused prominent nuclei condensation and significant DNA fragmentation, a phenomenon association with programmed cell death. Cholesterol oxides were able to potentiate the bacterial lipopolysaccharide (LPS)-induced nitric oxide production to various degrees. Consistent with this finding, Northern blot analysis indicated that 25-OH cholesterol potentiated the LPS-induced nitric oxide synthase RNA levels. The cytotoxicity of 25-OH-cholesterol could be prevented by methyl-beta-cyclodextrin, a glucose polymer known to cause cholesterol oxide efflux from cells. While much attention has been focused on the cytotoxicity of cholesterol oxides on immune cells within the blood, including lymphocytes and macrophages, the results from this study indicated for the first time that these agents are toxic to microglial cells derived from the central nervous system. PMID- 9731220 TI - Molecular cloning, genomic organization and developmental regulation of a novel receptor from Drosophila melanogaster structurally related to gonadotropin releasing hormone receptors for vertebrates. AB - After screening the data base of the Berkeley Drosophila Genome Project with a sequence coding for the transmembrane region of a G protein-coupled receptor, we found that Drosophila might contain a gene coding for a receptor that is structurally related to the Gonadotropin-Releasing Hormone (GnRH) receptors from vertebrates. Using the polymerase chain reaction, with Drosophila cDNA as a template, and oligonucleotide probes coding for the presumed exons of this gene, we were able to clone the cDNA coding for this receptor. The transmembrane region of the receptor shows 36% amino acid residue identity with the transmembrane region of the catfish and 31% amino acid residue identity with that of the rat GnRH receptor. The Drosophila receptor gene contains six introns, whereas the rat gene contains two: one intron in the Drosophila gene occurs at exactly the same position and has the same intron phasing as one intron in the rat gene, suggesting that the Drosophila and mammalian GnRH receptor genes are evolutionarily related. Northern blot analyses show that the Drosophila receptor gene is progressively expressed during larval development with a prominent maximum at the 3rd instar larval stage. Pupae contain low amounts of receptor mRNA, while adult flies contain higher levels, with males having about five times more receptor mRNA than females flies. Southern blot analyses show that Drosophila contains only one copy of the receptor gene, which is located at position 27A2-B1 of chromosome 2. This paper is the first report on the molecular cloning of a member of the GnRH receptor family from invertebrates. PMID- 9731221 TI - Lack of direct connection between arachidonic acid release and prostanoid synthesis upon differentiation of U937 cell. AB - The changes in AA incorporation and release as well as prostanoid synthesis upon differentiation of human premonocytic cell line, U937, induced by three functionally diverse agents--phorbol ester (TPA), dimethyl sulfoxide (DMSO), and retinoic acid (RA) have been investigated. The rate of AA incorporation into the cells remained unchanged whereas a 3- to 6-fold increase in AA release upon stimulation with Ca(2+)-ionophore A23187 as compared to undifferentiated cells was observed. While undifferentiated cells were incapable to metabolise AA via the cyclooxygenase pathway all three types of differentiated U937 cells produced TxB2 and PGE2. Only TPA-differentiated cells responded with a 6-fold increase of prostanoid synthesis after A23187 stimulation, whereas in DMSO-differentiated cells prostanoid synthesis was slightly stimulated by A23187 and in RA differentiated cells it was not stimulated at all. Thus, agonist-induced prostanoid synthesis in differentiated cells is dependent on the nature of differentiating agent and does not correlate with AA liberation. PMID- 9731222 TI - Propofol reacts with peroxynitrite to form a phenoxyl radical: demonstration by electron spin resonance. AB - Peroxynitrite (ONOO-), resulting from the reaction of nitric oxide with superoxide anion, is a powerful oxidant produced in activated macrophages, during ischemia-reperfusion processes as well as in neurodegenerative disorders. This study investigated the reaction of the anesthetic agent propofol (PPF) with ONOO , using electron spin resonance (ESR) and UV-visible spectrometry. Peroxynitrite was synthetized either from acidified hydrogen peroxide and nitrite, or from sodium azide and ozone. The addition of ONOO- to PPF in alkaline solution (pH 12) allowed to detect a, short lifetime, ESR signal corresponding to a phenoxyl radical. This finding was confirmed by a UV-visible study, resulting in the appearance of 427 nm peak and the disappearance of the peak located at 239 nm. The 291 nm peak remained unchanged. The identification of the end-product of the reaction of PPF with ONOO- needs further investigations. PMID- 9731223 TI - Dopamine formation from tyramine by CYP2D6. AB - Dopamine is formed form L-tyrosine by tyrosine hydroxylase and aromatic L-amino acid decarboxylase. In addition to this pathway, however, the formation of catecholamines, including dopamine, from trace amines such as tyramine by hepatic microsomes has been demonstrated. In this study, we investigated the formation of dopamine from trace amines, using human hepatic microsomes and human cytochrome P450 (CYP) isoforms expressed in yeast. Among the 11 isoforms of human CYP expressed in yeast, CYP2D6 was the only isoform exhibiting strong ability to convert p-tyramine and m-tyramine to dopamine. In studies with human hepatic microsomes, the hydroxylation of tyramine to dopamine was inhibited by bufuralol, a typical substrate for CYP2D isoforms, and anti-CYP2D1 antiserum. This is the first report showing that CYP2D is capable of converting tyramine to dopamine. The Km values of CYP2D6, expressed in yeast, for p-tyramine and m-tyramine were 190.1 +/- 19.5 microM and 58.2 +/- 13.8 microM, respectively. Tyramine is an endogenous compound which exists in the brain as a trace amine but is also an exogenous compound which is found in foods such as cheese and wine. Our results suggest that dopamine is formed from endogenous and/or exogenous tyramine by this CYP2D isoform. PMID- 9731224 TI - Unusual effect of high hydrostatic pressure on basic phospholipase A2 from venom of Agkistrodon Halys Pallas. AB - The pressure effect on basic phospholipase A2 (BPLA2) from the venom of Agkistrodon Halys Pallas from the Zhe-Jiang province of China was studied by fluorescence spectroscopy from 0.1 to 650 MPa. It was found that the pressure effect on the tryptophan residue fluorescence emission spectra of the enzyme were was significantly different in two pressure ranges: from 0.1 to 400 MPa and from 400 to 650 MPa respectively. For increasing pressure, the spectrum shifted to the red in the lower pressure range and to the blue in the higher pressure range. Whereas the red shift could be ascribed to the intrinsic pressure dependence of the fluorophore (trp), the blue shift indicated a pressure induced protein conformational change toward a structure where the single tryptophan is in a less polar environment, suggesting its burying deeper inside the protein. This is the first time that such a phenomenon has been observed. Generally, high pressure denaturation of proteins leads to a red shift of tryptophan fluorescence. It was also found that the break point in pressure at which the blue shift began was dependent both on temperature and on the presence of Ca+2 ion, but not on the protein concentration. Experiments at different BPLA2 concentrations and light scattering under pressure indicated that the blue shift was not caused by protein aggregation under high pressure. PMID- 9731225 TI - Efficient and stable gene transfer following microinjection into nuclei of synchronized animal cells progressing from G1/S boundary to early S phase. AB - We examined the possible phase(s) of the cell cycle in which a foreign gene can be stably transferred to animal cells. DNA of the plasmid pSV2neo containing the neomycin-phosphotransferase gene was microinjected into the nuclei of NIH/3T3 cells synchronized by serum starvation and aphidicolin treatment. The frequency of neo(r)-transformation (expressed as a percentage of microinjected cells) was 6% at the G0 phase and increased with progression of the cell cycle to reach a peak of 76% at the G1/S boundary. When the cells started their growth from the G1/S following release from aphidicolin, the frequency increased or decreased in the parallel with the BrdU-labeling index. Furthermore we developed a simplified method in which asynchronously growing cells were treated with aphidicolin at 10 micrograms/ml fro 16 hrs without serum starvation and subjected to microinjection, and their growth was further induced in aphidicolin-free medium. Using five cell lines (BALB/3T3, BALB/MK-2, NRK, CHO-K1, and HeLa) and one primary culture of chicken embryo fibroblasts (CEF), a 3- to 7-fold increase in the frequency of neo(r)-transformation was consistently detected in aphidicolin treated cells, compared to non-treated asynchronous cultures. The present study indicates that synchronized animal cells progressing from the G1/S boundary to the early S phase integrate the PSV2neo DNA into their chromosomes with high efficiency. PMID- 9731226 TI - Ouabain suppresses ATP elevation in response to fuel secretagogues in pancreatic islets. AB - The alterations of the ATP concentration in response to fuel secretagogues such as glucose, glyceraldehyde, and ketoisocaproate (KIC) were investigated in a single islet. The intraislet ATP concentration was transiently elevated and then decreased to a level slightly higher than basal. To asses the ATP content under conditions of reduced ATP consumption, the Na-K pump blocker ouabain was used. The elevation of ATP concentration was found unexpectedly to be suppressed under ouabain in the islet, even when incubated with any of the secretagogues. High glucose did not elevate the intracellular creatine phosphate during incubation with ouabain. Since the suppression rate for the intraislet ATP elevation was considerably smaller with KIC than with glyceraldehyde and glucose, we conclude that ouabain inhibits ATP production, at least in part, in the glycolytic pathway through a feedback mechanism. PMID- 9731227 TI - The protective effect of 17 beta-estradiol on vasomotor responses of aorta from cholesterol-fed rabbit is reduced by inhibitors of superoxide dismutase and catalase. AB - The involvement of endogenous antioxidant enzymes in the protective effect of 17 beta-estradiol against hypercholesterolemia was evaluated on aorta from cholesterol-fed rabbits treated with estradiol. 17 beta-Estradiol, more potent than alpha-tocopherol, restored the acetylcholine-induced relaxation, which was impaired by cholesterol chow, and enhanced the vasorelaxation induced by sodium nitroprusside. Diethyldithiocarbamate (5 mM), a superoxide dismutase (SOD) inhibitor, reduced relaxation to acetylcholine down to the level obtained in cholesterol-fed rabbits not treated with estrogen. This inhibition was dose dependently reversed by addition of exogenous SOD. Aminotriazole (5 mM), a catalase inhibitor, reduced slightly this response, which was not reversed by exogenous catalase. A similar concentration of diethyldithiocarbamate prevented the potentiation of response to sodium nitroprusside induced by estrogen, whereas aminotriazole had no effect. These results suggest that, on aorta from cholesterol-fed rabbit, superoxide dismutase and catalase are involved in the protective effect of estrogen on the vasomotor response to acetylcholine, but only superoxide dismutase participates in response to sodium nitroprusside. PMID- 9731228 TI - Downregulation of the Klotho gene in the kidney under sustained circulatory stress in rats. AB - We recently reported the isolation of the klotho gene, which in predominantly expressed in the kidney and involved in human aging phenotypes. In our previous studies, we demonstrated that the Klotho protein or its metabolites may possibly function as humoral factor(s) and protect against endothelial dysfunction because acetylcholine-mediated NO production in arteries was impaired in heterozygous klotho deficient mice (kl/+). However, the pathophysiological significance of the Klotho protein has not been clarified yet. In the present study, we examined expression of the klotho gene in the kidney of the following rat models for human diseases: (1) spontaneously hypertensive rat, (2) deoxycorticosterone acetate salt hypertensive rat, (3) 5/6 nephrectomized rat, (4) non-insulin-dependent diabetes mellitus rat (the Otsuka Long-Evans Tokushima Fatty rat), and (5) rat with acute myocardial infarction. The expression levels of klotho mRNA in the kidney in these models were significantly lower than controls except for MI rats. This is the first report showing the expression of the klotho gene in the kidney is regulated under sustained circulatory stress such as long-term hypertension, diabetes mellitus, and chronic renal failure. PMID- 9731229 TI - cdc2 kinase-mediated phosphorylation of splicing factor SF2/ASF. AB - SR proteins are a family of splicing factors which are important components of spliceosomes. Recent studies suggested that phosphorylation of SR protein might be a key event for the regulation of pre-mRNA splicing and is prevalent in metaphase cells. To investigate the role of cdc2 kinase in cell cycle-dependent phosphorylation of SR protein, we examined its phosphorylation of SF2/ASF, a representative SR protein. SF2/ASF was phosphorylated both by recombinant cdc2 kinase, a cdc2-cyclin B complex, and by cdc2 kinase immunoprecipitated from G2/M phase HeLa cells. In vitro phosphorylation and phosphopeptide mapping of several mutant proteins revealed that cdc2 kinase specifically phosphorylates the RS domain of SF2/ASF with serines 227, 238 and presumably 199 as major phosphorylation sites. These findings suggest the possibility that cdc2 kinase takes part in the cell cycle-dependent phosphorylation of SR protein which regulates the function of spliceosomes. PMID- 9731230 TI - Cloning of a novel cDNA expressed during the early stages of fracture healing. AB - Using differential mRNA display (DD-PCR), a novel cDNA, FxC1 (Fracture Callus 1) was isolated from the early stages of a healing fractured femur. Utilizing 5' RACE PCR, a 598-bp full-length cDNA was obtained for FxC1 that contains an open reading frame (ORF) of 243 bp, encoding for an 80 amino acid protein. Within this ORF, a leucine zipper motif was present. In vitro transcription/translation of the full-length cDNA generated the expected 9-kDa protein. Northern analysis reveals that this gene is expressed in calluses harvested from post-fracture day 5, 7 and 10, as well as in several other tissues and bone-derived cell lines. During the differentiation of MC3T3 cells along the osteoblast lineage, FxC1 expression increases 3- to 4-fold during the production and deposition of matrix proteins, suggesting a possible role for this protein in cell differentiation. PMID- 9731231 TI - Expression of cartilage-specific functional matrix chondromodulin-I mRNA in rabbit growth plate chondrocytes and its responsiveness to growth stimuli in vitro. AB - Cartilage-specific functional matrix chondromodulin-I (ChM-I) is a 25 kDa glycoprotein purified from fetal bovine epiphyseal cartilage which stimulates the growth of rabbit chondrocytes and the colony formation of the cells in agarose. In the present study, we isolated rabbit ChM-I precursor CDNA by reverse transcription-polymerase chain reactions. Northern blot analysis revealed that the expression of ChM-I mRNA occurred in a tissue-specific manner in cartilage. Moreover, the ChM-I mRNA level was markedly changed in response to growth and differentiation stimuli in primary cultured chondrocytes. Among others, fibroblast growth factor-2, transforming growth factor-beta, and parathyroid hormone related peptide each markedly down-regulated the expression of ChM-I mRNA in chondrocytes. These results indicated that the expression ChM-I was placed under the dynamic control of local growth and differentiation factors. PMID- 9731232 TI - Ectopic expression of a chimeric colony-stimulating factor-1/TrkB-receptor promotes CSF-1-dependent survival of cultured sympathetic neurons. AB - The regulation of the density of innervation and the promotion of survival of neurons are the original effects depending on neurotrophins. Here we analyse such effects evoked by trkB tyrosine kinase in transfected PC12 cells and transfected sympathetic neurons. In order to exclude the previously described modulation of trk kinase activity by the extracellular activation of the low-affinity p75 neurotrophin receptor, we applied a chimeric receptor approach: The extracellular domain of colony-stimulating factor-1 (CSF-1) receptor was fused to the transmembrane and cytoplasmic domain of the trkB tyrosine kinase receptor, allowing its selective activation by the heterologous ligand. Protein expression and CSF-1-induced tyrosine phosphorylation of the chimeric receptor protein was demonstrated in transfected COS cells. After stable transfection into nerve growth factor (NGF)-responsive PC12 cells, CSF-1 mediated the K252a-sensitive induction of fiber outgrowth. Furthermore, we were able to show by heterologous expression of the chimeric receptor, that activation of trkB tyrosine kinase activity is sufficient to promote survival of neurotrophin deprived sympathetic neurons. PMID- 9731233 TI - Identification of the Gbeta5-RGS7 protein complex in the retina. AB - The G protein beta subunit G beta 5 deviates significantly from the four other members of the G beta family in amino acid sequence, unique expression pattern (only in the CNS), and cytosolic localization. To identify the members of the G beta 5-mediated signaling pathway, we purified the native protein complex containing G beta 5 from the cytosolic fraction of bovine retina. Analysis of the isolated complex revealed that G beta 5 is tightly associated with RGS7, a member of the superfamily of negative regulators of G protein signaling. This finding, for the first time, demonstrates an interaction between a G beta subunit and an RGS protein. G beta 5 was not detected in the outer segments of photoreceptor cells, suggesting that the cytosolic G beta 5-RGS7 complex is not directly involved in phototransduction. PMID- 9731234 TI - Fine structural and functional consequences of deglycosylation of the platelet adhesion receptor GPIb-IX (CD 42b). AB - To investigate the role of the glycosylation of the platelet receptor glycoprotein Ib (GPIb, CD 42b), platelets and purified GPIb were deglycosylated by neuraminidase, O- and N-glycosidases. N-deglycosylation and neuraminic-acid cleavage had little effect on ristocetin and botrocetin-induced platelet agglutination. However, O-deglycosylation reduced the response by approximately 50%, and total deglycosylation (the combination of all three glycosidases) fully abolished the response to ristocetin. Interestingly, binding of von Willebrand Factor (vWF) to purified GPIb in the presence of ristocetin and botrocetin in a standardized microtiter plate assay was not altered by partial or even by total deglycosylation. Electron microscopy indicated that the normally stretched approximately 50 nm long molecule was approximately 32 nm after N deglycosylation, approximately 20 nm after O-deglycosylation, and reduced in a approximately 15 nm long collapse by total deglycosylation. These results suggest that deglycosylation has major structural impacts on GPIb, strongly impairing platelet-vWF interactions; however, vWF binding to isolated GPIb remains unaffected. PMID- 9731235 TI - Remodeling of HDL by phospholipid transfer protein: demonstration of particle fusion by 1H NMR spectroscopy. AB - There is evidence that phospholipid transfer protein (PLTP) can increase reverse cholesterol transport by inducing favorable subclass distribution in the high density lipoprotein (HDL) fraction. This includes generation of initial cholesterol acceptor particles, pre beta-HDL, and of enlarged particles that are rapidly cleared from the circulation. However, partly because of methodological difficulties, the mechanisms behind the PLTP-mediated interconversion of HDL particles are not fully understood. In this communication, we describe the use of a novel methodology, based on 1H NMR spectroscopy, to study the PLTP-induced size changes in the HDL particles. In accordance with native gradient gel electrophoresis, the 1H NMR data revealed a gradual production of enlarged HDL particles in the HDL3+ PLTP mixtures. In addition, according to a physical model for lipoprotein particles, relating the frequency shifts observable with NMR to the size of the lipoprotein particles, the NMR data demonstrated that PLTP mediated HDL remodeling involves fusion of the HDL particles. PMID- 9731236 TI - The chaperone-like alpha-crystallin forms a complex only with the aggregation prone molten globule state of alpha-lactalbumin. AB - The chaperone-like alpha-crystallin prevents aggregation of several proteins by interacting with their non-native states. Alpha-Lactalbumin adopts different non native states under different experimental conditions. We have investigated the interaction of alpha-crystallin with three non-identical non-native states, using fluorescence, circular dichroism, and gel filtration chromatography. The compact molten globule state of apo-alpha-lactalbumin in tris buffer does not interact with alpha-crystallin. The expanded, flexible molten globule-like state of reduced apo-alpha-lactalbumin (formed at pH 7.2) also does not interact with alpha-crystallin. Only the aggregation-prone non-native state of reduced apo alpha-lactalbumin formed at pH 6.0 interacts with alpha-crystallin to form a stable complex. The alpha-crystallin bound reduced apo-alpha-lactalbumin exhibits properties similar to those of a molten globule. Our results show that alpha crystallin interacts only with the aggregation prone molten globule state of reduced apo-alpha-lactalbumin but not with the other non-aggregating molten globule states of the protein. PMID- 9731238 TI - Zonal induction of mixed lineage kinase ZPK/DLK/MUK gene expression in regenerating mouse liver. AB - ZPK/DLK/MUK is a serine/theronine kinase believed to be involved in the regulation of cell growth and differentiation. To further explore the suggested participation of ZPK/DLK/MUK in this process, we examined the expression and cellular localization of ZPK/DLK/MUK mRNA in regenerating mouse liver following partial hepatectomy by ribonuclease protection assay and in situ hybridization. The steady-state level of APK/DLKMUK mRNA was very low in normal and sham operated mouse livers, whereas a marked and transient increase was observed in the regenerating liver. While ZPK/DLK/MUK mRNAs were rarely detected in hepatocytes from all zones of the normal liver, hepatocytes of regenerating liver exhibit a gradient of expression ranging from low in the periportal zone, to intermediate in the mid-zone, to high in the pericentral zone. These findings demonstrate a transient stimulation of ZPK/DLK/MUK gene expression that correlates with the growth response of hepatocyte subpopulations in regenerating liver. PMID- 9731237 TI - Novel anti-respiratory syncytial(RS) viral compounds: benzodithiin derivatives. AB - Benzodithiin derivatives are highly potent and specific inhibitors of respiratory syncytial virus (RSV) replication in vitro. The most potent and selective congener of a benzodithiin derivative is 1,4-dihydro-2,3-benzodithiin(RD3-0028). According to the modified 3-(4,5-dimethylthiazole-2-yl) 2,5-diphenyl tetrazolium bromide (MTT) assay developed in our laboratories, this compound has a 50% effective concentration of 4.5 microM and a 50% cytotoxic concentration of 271.0 microM, which is superior to that of ribavirin. This compound also inhibits RSV strain subgroups A and B and clinical isolates. RD3-0028, however, does not inhibit the replication of influenza A virus, measles virus, herpes simplex virus types 1 and 2, or human cytomegalovirus. Two other benzodithiin derivatives [1,4 dihydro-6-methyl-2,3-benzodithiin (RD3-0270) and 1,4-dihydro-5-methyl-1-2,3 benzodithiin (RD3-0284)] also inhibit RSV replication at a selectivity index greater a factor of 20. These results suggest that the benzodithiin skeleton is an important structure for inhibitory activity against RSV replication. PMID- 9731239 TI - Structural and functional consequences of substitutions at the Pro108-Arg14 hydrogen bond in bovine adrenodoxin. AB - Elimination of Pro108 in bovine adrenodoxin is known to result in the formation of a misfolded protein that is not able to incorporate a [2Fe-2S] cluster and rapidly degrades upon expression in E. coli. However, no experimental explanation for this phenomenon has been demonstrated so far. Using the recently obtained 3D structure of the truncated mutant Adx(4-108) we have studied the reasons of the protein stabilization by the proline residue by means of site-directed muta genesis. Two main results have been obtained (i) the conserved hydrogen bond Pro108-Arg14, that connects different structural domains of Adx, contributes 6 kJ/mol into the protein stability and (ii) the presence of proline at position 108 provides a low conformational entropy of the unfolded state, supporting a gain in the Gibbs energy of 5.4 kJ/mol at 37 degrees C. PMID- 9731240 TI - Effects of hydroxyl radical and sulfhydryl reagents on the open probability of the purified cardiac ryanodine receptor channel incorporated into planar lipid bilayers. AB - We examine the effect of hydroxyl radical on the ion permeability of the ryanodine receptor, a calcium releasing channel of sarcoplasmic reticulum. The cardiac ryanodine receptor, purified from pig heart, was reconstituted to proteoliposomes and then incorporated into a planar bilayer membrane. A single channel activity with a conductance of 724 pS in 900/200 mM (cis/trans) KC1 and an ion selectivity of PK:PCl = 1:0.08 was observed. These characteristics are similar to those observed by the incorporation of the channel through sarcoplasmic reticulum vesicles. Hydroxyl radicals chemically generated by the reaction of H2O2 and Cu(ethylenediamine)2 at the cis compartment increased the open probability of the channel. Treatment with SH oxidizing reagents from the cis compartment also increased the open probability, and dithiothreitol, a SH reducing agent, reversed the effect. These findings suggest that hydroxyl radicals react with some SH groups of the ryanodine receptor and increase the Ca2+ release from sarcoplasmic reticulum through the ryanodine receptor. PMID- 9731241 TI - Identification of human beta-defensin-2 in respiratory tract and plasma and its increase in bacterial pneumonia. AB - Human beta-defensin-2 (hBD-2), a novel antimicrobial peptide, was originally isolated from human skin. We found that synthetic hBD-2 has high bactericidal activity against Escherichia coli under conditions nearly the same as in human bronchial airway surface liquid. We prepared an antiserum against hBD-2 and established a highly sensitive radioimmunoassay (RIA). Reverse-phase high performance liquid chromatography coupled with the RIA showed that hBD-2 in patients with human lung, bronchoalveolar lavage flid, and plasma. The plasma concentration of hBD-2 in patients with bacterial pneumonia was 32.1 +/- 3.7 fmol/ml (mean +/- SE), 3.9-fold that of normal individuals. By reverse transcription-polymerase chain reaction, the hBD-2 gene transcript was detected in the respiratory epithelial surface of human lung. Human beta-defensin-2 seems to function in airway mucosal defense. Our findings provide a clue to elucidate its pathophysiological significance in respiratory infection. PMID- 9731242 TI - Cefdinir--a new oral cephalosporin. PMID- 9731243 TI - Risedronate for Paget's disease of bone. PMID- 9731244 TI - New FDA rule affects drug labeling. PMID- 9731245 TI - In support of current vaccination protocols. PMID- 9731246 TI - Animal populations may not be out of control. PMID- 9731247 TI - Comments regarding retrospective study on horses with fungal keratitis. PMID- 9731248 TI - What is your diagnosis? Oblique fracture of the caudal half of the transverse process of the fourth cervical vertebra. PMID- 9731249 TI - ECG of the month. Atrial fibrillation with respiratory-associated atrioventricular cyclical blockade in a dog. PMID- 9731250 TI - Anesthesia case of the month. Complications during surgery for a spinal fracture in a dog. PMID- 9731251 TI - Animal behavior case of the month. A dog was evaluated because of severe aggression directed exclusively toward one of its owners. PMID- 9731252 TI - Role of long-chain polyunsaturated n-3 fatty acids in the development of the nervous system of dogs and cats. PMID- 9731253 TI - Mobile veterinary clinics. PMID- 9731254 TI - American College of Veterinary Anesthesiologists' position paper on the treatment of pain in animals. PMID- 9731255 TI - American Veterinary Dental College's position statement regarding veterinary dental health care providers. PMID- 9731256 TI - Comparison of gonioscopy and ultrasound biomicroscopy for evaluating the iridocorneal angle in dogs. AB - OBJECTIVE: To compare iridocorneal angle grading systems on the basis of gonioscopy and ultrasound biomicroscopy (UBM). DESIGN: Original cross-sectional observational study. ANIMALS: 22 dogs. PROCEDURE: Gonioscopy, goniophotography, and UBM were performed on 38 eyes from dogs without clinical evidence of glaucoma in the eyes examined. RESULTS: Predominant gonioscopic grades derived from goniophotographs were considered normal (n = 26) and mild (12). Ultrasound biomicroscopy angle measurements ranged from 16 to 38 degrees (mean +/- SD, 26.2 +/- 4.5 degrees). Ciliary clefts depicted on UBM images were graded as open (n = 4), compact/narrow (23), and closed (11). Significant differences were not found between UBM-derived ciliary cleft grades and goniophotography-derived dysgenesis grades, nor between UBM-derived ciliary cleft grades and subjective gonioscopic grades. CLINICAL IMPLICATIONS: Because gonioscopy allows evaluation of the anterior face of the ciliary cleft, whereas UBM provides cross-sectional information of the iridocorneal filtration angle, UBM may yield more information regarding pathogenesis and prognosis of, and preferred management approaches to, glaucoma. Ultrasound biomicroscopy may also be useful as a predictor of glaucoma or to diagnose early stages of glaucoma. PMID- 9731257 TI - Effect of distance traveled and speed of racing on body weight and serum enzyme activity of sled dogs competing in a long-distance race. AB - OBJECTIVE: To determine the relationship of serum biochemical values and change in body weight with finishing status (retired from or finished the race), finishing order of a team, and distance traveled for dogs participating in a long distance sled dog race. ANIMALS: 262 of 848 dogs that participated in the 1995 Iditarod Trail sled dog race. DESIGN: Prospective study. PROCEDURE: Body weight was recorded for 261 dogs before the race and again when these dogs retired from or completed the race. Using a nonrandom convenience sample of participating dogs, blood samples were obtained from 151 dogs that retired from the race and 111 dogs that completed the race. RESULTS: Serum biochemical indices of skeletal muscle damage were significantly higher in dogs retiring during the first 500 miles of the race than in dogs retiring in the last 638 miles or finishing the race. Serum sodium concentration was less than the reference range in a significantly greater proportion of dogs that retired from the race than of dogs that completed the race. There was little relationship between finishing order and serum biochemical values. Dogs completing the race lost a mean of 8.9% of body weight, and amount of weight lost was not related to finishing order. CLINICAL IMPLICATIONS: Results indicated that exertional rhabdomyolysis develops more often in dogs that retire during the initial 500 miles of a long-distance race, compared with dogs that complete the race. There is no detectable relationship between the speed with which the race is run (finishing order) and body weight loss or serum biochemical values. PMID- 9731258 TI - Retinal degeneration associated with vitamin E deficiency in hunting dogs. AB - A group of Walker Hounds and Beagles that were fed a diet of table scraps were examined because of slow, progressive loss of vision. Clinical and microscopic features of the disease were correlated to the dogs' micronutrient status. Sensory retinal degeneration, predominantly in the central tapetal fundus, was found in all dogs, and severity of changes varied with age of the dog. Plasma, serum, and tissue concentrations of vitamin E were low in affected dogs (10 to 40% of control values). Lipofuscin accumulation was found on microscopic examination in retinal pigment epithelium, smooth muscle cells of the intestinal tract, and neurons of the CNS. Findings were consistent with nutritional vitamin E deficiency and oxidative injury to photoreceptors of the retina. Changes in these dogs were similar to those described for central progressive retinal atrophy and equine lower motor neuron disease, suggesting these diseases may share a common pathogenesis to vitamin E deficiency. PMID- 9731259 TI - Open-heart correction of tetralogy of Fallot in an acyanotic dog. AB - Tetralogy of Fallot was diagnosed in an acyanotic 11-month-old dog. Predicted pressure gradient across the pulmonic valve, as assessed by use of continuous wave Doppler echocardiography, was 94.5 mm Hg. Bidirectional shunting was identified by means of selective angiography. Open-heart correction was performed, using a transatrial approach with limited ventriculotomy and cardiopulmonary bypass. The hypertrophied infundibulum was resected, the ventricular septal defect was closed primarily, and a transannular pericardial patch graft was applied. Pressure gradients across the pulmonic valve were 52.9 and 22.8 mm Hg 2 weeks and 4 months after surgery, respectively. Advances in cardiopulmonary bypass, anesthetic management, and use of the transatrial approach may improve the success of open-heart correction of tetralogy of Fallot in dogs. PMID- 9731260 TI - Blastomycosis in dogs: 115 cases (1980-1995). AB - OBJECTIVE: To characterize diagnostic results, treatment, and outcome of dogs with blastomycosis during a 15-year period in Louisiana. DESIGN: Retrospective case series. ANIMALS: 115 dogs with blastomycosis. PROCEDURE: Medical records were reviewed for dogs with blastomycosis examined between 1980 and 1995. Additional data were collected from the state veterinary diagnostic laboratory, via telephone interviews of owners, and by use of a random survey of the hospital population. RESULTS: Blastomycosis was detected mainly in young, large-breed dogs. Proximity to a body of water was a significant risk factor for affected dogs. Most dogs were affected in January and August through October. Clinical signs and results of physical examination reflected the multisystemic nature of the disease. Commonly affected systems included the respiratory tract and lymphatic, ocular, and cutaneous systems. Nodular interstitial and interstitial patterns were common findings on thoracic radiographs. Cytologic examination was successful in identifying organisms in samples from vitreous, skin, and lymph nodes. Similar results were achieved for dogs treated with a combination of amphotericin B and ketoconazole, compared with dogs treated with itraconazole. CLINICAL IMPLICATIONS: Results of this study should assist veterinarians with the recognition and management of blastomycosis in dogs. Blastomycosis should be considered as a differential diagnosis for large-breed dogs that live close to a body of water in areas in which the disease is endemic or in dogs with a history of being transported to endemic areas that subsequently develop signs of pulmonary, ocular, lymphatic, or cutaneous disease. Treatment with itraconazole was as effective as treatment with a combination of amphotericin B and ketoconazole. PMID- 9731261 TI - Clinical, clinicopathologic, radiographic, and ultrasonographic abnormalities in dogs with fatal acute pancreatitis: 70 cases (1986-1995). AB - OBJECTIVE: To determine clinical, clinicopathologic, radiographic, ultrasonographic, and coagulation abnormalities in dogs in which acute pancreatitis was fatal. DESIGN: Retrospective study. ANIMALS: 70 dogs. PROCEDURE: History, clinical signs, and physical examination findings at the time of initial evaluation at the veterinary teaching hospital; results of pretreatment laboratory tests, abdominal radiography, and ultrasonography; and histologic abnormalities were obtained from medical records. RESULTS: Clinical signs included anorexia, vomiting, Weakness, diarrhea, polyuria and polydipsia, neurologic abnormalities, melena, weight loss, hematemesis, and passage of frank blood in feces. At the time of initial examination at the veterinary teaching hospital, 68 (97%) dogs were dehydrated, 18 (26%) were icteric, 22 (32%) were febrile, 40 (58%) had signs of abdominal pain, and 30 (43%) were classified as overweight or obese. Most dogs had leukocytosis, neutrophilia with a left shift, and thrombocytopenia. Various serum biochemical abnormalities were identified, including hypoglycemia, azotemia, hypercalcemia and other electrolyte abnormalities, hypoalbuminemia, high hepatic and pancreatic enzyme activities, hypercholesterolemia, and lipemia. For 17 of 28 (61%) dogs, results of coagulation function tests were abnormal. Results of abdominal ultrasonography and radiography were consistent with a diagnosis of acute pancreatitis in 23 of 34 (68%) and 10 of 41 (24%) dogs, respectively. For 2 dogs, results of abdominal ultrasonography were not suggestive of acute pancreatitis, but results of abdominal radiography were. CLINICAL IMPLICATIONS: Clinical signs and results of clinicopathologic tests are inconsistent. Abdominal ultrasonography may be valuable in the diagnostic evaluation of dogs suspected to have acute pancreatitis. PMID- 9731262 TI - Herpesvirus myeloencephalopathy in horses: 11 cases (1982-1996). AB - OBJECTIVE: To determine results of CSF analysis in horses with equid herpesvirus myeloencephalopathy (EHM) and to determine whether results of CSF analysis were associated with outcome. DESIGN: Retrospective study. ANIMALS: 11 horses. PROCEDURE: Medical records of all horses admitted to the veterinary teaching hospital between February 1982 and March 1996 in which EHM was diagnosed were reviewed. RESULTS: 7 horses were < or = 4 years old; 8 were admitted during January, February, or March. Six horses were febrile prior to admission, but none was febrile on the day of admission. Five horses had been stabled with other horses that had clinical signs of neurologic disease. All horses had had an acute onset of hind limb ataxia and paresis. Cranial nerve deficits were detected in 3 horses. Cerebrospinal fluid samples were collected on the day of admission from 10 horses. Protein concentration was high in 8 horses; nucleated cell count was normal in 8. Protein concentration and nucleated cell and RBC counts were not significantly different between horses that survived and horses that were euthanatized. Six horses were euthanatized, and 5 survived. All of the horses that survived remained standing or were able to stand with minimal assistance. CLINICAL IMPLICATIONS: High CSF protein concentration and normal or only slightly high CSF nucleated cell count are common in horses with EHM; however, results of CSF analysis were not associated with outcome. Horses with EHM that become recumbent have a poor prognosis for survival. PMID- 9731264 TI - Determination of the association between Neospora caninum infection and reproductive performance in beef herds. AB - OBJECTIVE: To determine seroprevalence of Neospora caninum infection in beef herds and the association between serologic status and rate of abortion, stillbirth, calf mortality, and reproductive failure. DESIGN: Longitudinal study. ANIMALS: 419 cows from 8 beef herds in central Alberta. PROCEDURE: 1,391 serum samples from a serum bank were analyzed, using ELISA, and results were compared, using logistic regression, with productivity data on individual cows obtained from a database established during a 4-year period. RESULTS: 30% of cows were seropositive at some point during the 4-year period. Risks of abortion (odds ratio [OR], 5.7) and stillbirth (OR, 2.8) in seropositive cows were significantly greater than in seronegative cows. Risks of being culled for any reason (OR, 1.9) or for reproductive failure (OR, 2.5) in seropositive cows were also significantly greater than in seronegative cows. Changes in titer with time in individual cows and a lack of association between serologic status of dam versus daughter suggest that postnatal transmission was possible in these herds. However, horizontal transmission did not appear to play a substantial role in abortions that occurred in these herds. CLINICAL IMPLICATIONS: Neosporosis should be investigated as a potential source of economic loss to the beef industry. PMID- 9731263 TI - Comparison of antibiotic administration in conjunction with supportive measures versus supportive measures alone for treatment of dairy cows with clinical mastitis. AB - OBJECTIVE: To determine whether antibiotic and supportive treatment would improve outcome for dairy cows with naturally developing clinical mastitis, compared with supportive treatment alone. DESIGN: Randomized controlled trial. ANIMALS: 124 cows in one herd with 172 episodes of clinical mastitis. PROCEDURE: Cows were examined at the onset of clinical mastitis, assigned a severity score, and randomly assigned to receive antibiotic (intramammary administration of cephapirin, i.v. administration of oxytetracycline, or both) and supportive treatment (administration of oxytocin, stripping of affected glands, and, in severely affected cows, administration of flunixin meglumine or fluids) or supportive treatment alone. Treatment was continued until 24 hours after signs of clinical mastitis resolved (clinical cure). Milk samples from affected glands were submitted for bacterial culture before initial treatment and every 2 weeks thereafter until the causative organism was no longer isolated (bacteriologic cure). RESULTS: When mastitis was caused by Streptococcus spp or coliform bacteria, clinical cure rate by the tenth milking was significantly higher if antibiotics were used. Bacteriologic cure rate at 14 days was significantly higher when antibiotics were used, particularly if mastitis was caused by Streptococcus spp. Cows receiving antibiotics developed fewer subsequent episodes of clinical mastitis during the 60 days after the initial episode of mastitis and had less severe clinical disease than cows that did not. CLINICAL IMPLICATIONS: Results suggest that, in herds in which mastitis is often caused by environmental bacteria, antibiotic and supportive treatment may result in a better outcome for cows with clinical mastitis than supportive treatment alone. PMID- 9731266 TI - A method for a simulation of continuous intracranial pressure curves. AB - The study introduces a method to simulate continuously an intracranial pressure (ICP) wave form. In a system analysis approach the intracranial compartment was viewed as a black box with arterial blood pressure (ABP) as an input signal and ICP as an output. A weight function was used to transform the ABP curve into the ICP curve. The output ICP waveform was generated using a weight function derived from the transcranial Doppler blood flow velocity (FV) and ABP curves. In order to establish the relationship between TCD characteristics and weight functions simultaneous recordings of FV, ABP, and ICP curves of a defined group of patients were used. A linear function between the TCD characteristics and the weight functions was obtained by calculating a series of multiple regression analyses. Given examples demonstrate the procedure's capabilities in predicting the mean ICP, the pulse and respiratory waveform modulations, and the trends of ICP changes. PMID- 9731265 TI - Evaluation of a portable clinical analyzer in a veterinary hospital setting. AB - OBJECTIVE: Evaluation of a portable clinical analyzer for determination of blood gas tensions, electrolyte and glucose concentrations, and Hct in a hospital setting. DESIGN: Prospective study. ANIMALS: 50 dogs, 50 cats, and 28 horses, all clinically normal. PROCEDURE: Blood samples were analyzed on a portable clinical analyzer to determine concentrations of sodium, potassium, chloride, BUN, glucose, and ionized calcium and values of Hct, pH, PCO2, and PO2. Values obtained were compared with those obtained from the same blood samples, using a standard automatic analyzer (serum sodium, potassium, chloride, BUN, and glucose concentrations), a cell counter (Hct), a blood gas analyzer (pH, PCO2, PO2), and a calcium-pH analyzer (ionized calcium). Bias (mean difference between values obtained on the same sample by different methods) and variability (SD of differences) were determined for all values. Data were also subjected to Deming regression analysis. RESULTS: Correlation coefficients were > 0.90 for all values except potassium and ionized calcium concentrations. Bias and variability were within clinically acceptable limits (+/- 2 SD) for all but potassium, ionized calcium, and glucose concentrations and Hct. Species-dependent variability was observed for glucose concentration and Hct. CLINICAL IMPLICATIONS: Most differences between values obtained with the portable clinical analyzer and standard clinical laboratory systems could be accounted for by differences in type of sample tested (blood vs serum). The portable clinical analyzer is suitable for point-of-care analysis in critical care situations and for routine blood biochemical analysis when extensive laboratory support is unavailable. PMID- 9731267 TI - The calculation of a confidence interval on the absolute estimated benefit for an individual patient. AB - Physicians need a method to predict the individualized absolute therapeutic benefit before deciding which therapy to prescribe to a given patient and the confidence intervals around this estimate. We have derived a method to predict the absolute individual therapeutic benefit in a previous work. In this paper, we present a Monte Carlo simulation to estimate the bias of prediction for an individual with certain characteristics and use a bootstrap method to compute its confidence intervals. Because the bootstrap approach does not depend upon the parametric assumption for the distribution of the prediction, it can be applied to situations where the parametric distribution is unknown. Over 35,000 cases of subjects at risk of cardiovascular events were available for analysis. Our results show the 95% confidence intervals for each individual. In a clinical setting, the use of this approach makes it possible to predict the absolute therapeutic benefit for each patient (the quantity of individual benefit) with sufficient precision. PMID- 9731268 TI - Enhancing the power of record linkage involving low quality personal identifiers: use of the best link principle and cause of death prior likelihoods. AB - The Heartstart Scotland study collects details of all resuscitation attempts carried out by the Scottish Ambulance Service. The linkage between records for Heartstart study subjects who died before admission to a hospital and the national file of death records maintained by the Registrar General for Scotland is described. The conditions under which the Heartstart data is collected make it inevitable that the personal identifying information on which linkage must rely tends to be relatively incomplete and of low accuracy. The linkage process was able to use the best-link principle to take maximum advantage of the fact that, because the Heartstart subjects involved had died, there was an extremely high a priori probability that they would be represented on the national deaths file. In addition, although no cause of death information was recorded on the Heartstart records, a priori expectations of the distribution of causes of death among linked death records were used. Despite these enhancements, however, clerical resolution of a proportion of the potential links generated by the automatic algorithm significantly improved the accuracy of the linkage. PMID- 9731269 TI - Comparative study of local and Karhunen-Loeve-based ST-T indexes in recordings from human subjects with induced myocardial ischemia. AB - In this work we studied ST-T complex changes in the ECG as result of induced ischemia. The principal aim was to determine whether global changes in the ST-T complex were more sensitive markers of ischemic alterations than those based on measurements of changes at specific locations on ST segment or T wave. High resolution ECGs from patients undergoing percutaneous transluminal coronary angioplasty in one of the major coronary arteries were analyzed to give a description of the period from the end of active depolarization (QRS complex) to the end of active repolarization (T wave). During artery occlusion traditional local measurements of the ST-T complex were compared to global measurements based on the Karhunen-Loeve transform. An ischemic change sensor parameter was estimated for each of the studied indexes showing that global measurements detected changes better in the repolarization period in a larger number of leads and with higher sensitivity (more than 85%) than was done using local measurements (sensitivity of 64% with ST level, 33% with T-wave maximum position, and 37% with T-wave maximum amplitude). Using these global indexes it was found that most cases of ST-segment changes were accompanied by T-wave changes (72% of patients). With the use of traditional indexes 23% of patients showed no changes in the repolarization period, whereas with global indexes this percentage decreased to 8%. Thus a global representation of the entire ST-T complex appears to be more suitable than local measurements when studying the initial stages of myocardial ischemia. PMID- 9731270 TI - Myocardial infarction--pinpointing the key indicators in the 12-lead ECG using data mining. AB - In this paper we describe how data mining techniques were used in order to pinpoint the key indicators for myocardial infarction in the electrocardiogram (ECG) by determining existing trends in a large data set. In order to provide a test bed for the data mining techniques a data mining tool was developed so that the effectiveness of various data mining techniques could be determined. The material consisted of 2730 ECGs recorded at an emergency department. A total of 517 ECGs were recorded on patients suffering acute myocardial infarction. The remaining ECGs were defined as control ECGs. A subset of the material was used to train the data mining tool. After training, the data mining tool was able to pinpoint the key ECG indicators for myocardial infarction in the test set (duration and amplitude of the Q wave and R duration in lead V2) and successfully determine which patients had suffered a heart attack. PMID- 9731271 TI - Desulfotomaculum halophilum sp. nov., a halophilic sulfate-reducing bacterium isolated from oil production facilities. AB - A halophilic endospore-forming, sulfate-reducing bacterium was isolated from an oilfield brine in France. The strain, designated SEBR 3139, was composed of long, straight to curved rods. It grew in 1-14% NaCl with an optimum at 6%. On the basis of morphological, physiological and phylogenetical characteristics, strain SEBR 3139 should be classified in the genus Desulfotomaculum. However, it is sufficiently different from the hitherto described Desulfotomaculum species to be considered as a new species. Strain SEBR 3139T (= DSM 11559T) represents the first moderate halophilic species of the genus Desulfotomaculum. The name Desulfotomaculum halophilum sp. nov. is proposed. PMID- 9731272 TI - Alcanivorax borkumensis gen. nov., sp. nov., a new, hydrocarbon-degrading and surfactant-producing marine bacterium. AB - During screening for biosurfactant-producing, n-alkane-degrading marine bacteria, six heterotrophic bacterial strains were isolated from enriched mixed cultures, obtained from sea water/sediment samples collected near the isle of Borkum (North Sea), using Mihagol-S (C14,15-n-alkanes) as principal carbon source. These Gram negative, aerobic, rod-shaped bacteria use a limited number of organic compounds, including aliphatic hydrocarbons, volatile fatty acids, and pyruvate and its methyl ether. During cultivation on n-alkanes as sole source of carbon and energy, all strains produced both extracellular and cell-bound surface-active glucose lipids which reduced the surface tension of water from 72 to 29 mN m-1 (16). This novel class of glycolipids was found to be produced only by these strains. The 16S rRNA gene sequence analysis showed that these strains are all members of the gamma-subclass of the Proteobacteria. Their phospholipids ester linked fatty acid composition was shown to be similar to that of members of the genus Halmonas, although they did not demonstrate a close phylogenetic relationship to any previously described species. On the basis of the information summarized above, a new genus and species, Alcanivorax borkumensis, is described to include these bacteria. Strain SK2T is the type strain of A. borkumensis. PMID- 9731273 TI - Phylogeny of Rhizobium galegae with respect to other rhizobia and agrobacteria. AB - PCR-RFLP with nine restriction enzymes was applied to the 16S and 23S rRNA genes of 42 rhizobial and agrobacterial strains to determine the phylogenetic position of Rhizobium galegae and increase the understanding of the evolution of ribosomal operons. The strains were selected based on previous phylogenetic studies. PCR primers were designed so that they amplified a 2.3 kb fragment of the 23S rRNA gene (excluding the B8 loop). Universal primers rD1 and fD1 were used to amplify the full-length 16S rRNA. The RFLP analysis resulted in 27 and 32 different restriction patterns for 16S and 23S, respectively. The RFLP patterns were transformed to genetic distances and dendrograms were constructed from the data using the unweighted pair group method with averages. The shapes of the dendrograms derived from the analysis of the 16S and 23S rRNA genes correlated well, with only a few strains having different positions. The 23S tree generally had deeper branching than the 16S tree, allowing better discrimination between species and strains. The combined data from the two analyses described 36 genotypes. The eight R. galegae strains formed a homogeneous cluster in all dendrograms. The RFLP analysis was confirmed by partial sequence analysis of the 16S rRNA gene (the first 800 bp), which correlated well with full-length 16S rRNA sequence analysis. The 16S data placed R. galegae near the genus Agrobacterium with Agrobacterium vitis as its nearest neighbour, whereas in the 23S and the combined dendrograms it showed closer affinity to the genus Rhizobium. PMID- 9731274 TI - Methanobacterium subterraneum sp. nov., a new alkaliphilic, eurythermic and halotolerant methanogen isolated from deep granitic groundwater. AB - Deep subterranean granitic aquifers have not been explored regarding methanogens until now. Three autotrophic methane-producing Archaea were isolated from deep granitic groundwater at depths of 68, 409 and 420 m. These organisms were non motile, small, thin rods, 0.1-0.15 micron in diameter, and they could use hydrogen and carbon dioxide or formate as substrates for growth and methanogenesis. One of the isolates, denoted A8p, was studied in detail. It grew with a doubling time of 2.5 h under optimal conditions (20-40 degrees C, pH 7.8 8.8 and 0.2-1.2 M NaCl). Strain A8p is eurythermic as it grew between 3.6 and 45 degrees C. It was resistant to up to 20 mg bacitracin l-1. The G + C content was 54.5 mol%, as determined by thermal denaturation. Phylogenetic studies based upon 16S rRNA gene sequence comparisons placed the isolate A8p in the genus Methanobacterium. Phenotypic and phylogenetic characters indicate that the alkaliphilic, halotolerant strain A8p represents a new species. We propose the name Methanobacterium subterraneum for this species, and strain A8p (= DSM 11074T) is the type strain. PMID- 9731275 TI - Characterization of tropical tree rhizobia and description of Mesorhizobium plurifarium sp. nov. AB - A collection of strains isolated from root nodules of Acacia species in Senegal was analysed previously by electrophoresis of total cell protein, auxanographic tests, rRNA-DNA hydridization, 16S rRNA gene sequencing, DNA base composition and DNA-DNA hybridization [de Lajudie, P., Willems, A., Pot, B. & 7 other authors (1994). Int J Syst Bacteriol 44, 715-733]. Strains from Acacia were shown to belong to two groups, Sinorhizobium terangae, and a so-called gel electrophoretic cluster U, which also included some reference strains from Brazil. Further taxonomic characterization of this group using the same techniques plus repetitive extragenic palindromic-PCR and nodulation tests is presented in this paper. Reference strains from Sudan and a number of new rhizobia isolated from nodules of Acacia senegal, Acacia tortilis subsp. raddiana and Prosopis juliflora in Senegal were included. As a result of this polyphasic approach, the creation of a new species, Mesorhizobium plurifarium, is proposed for a genotypically and phenotypically distinct group corresponding to the former cluster U and containing strains isolated from Acacia, Leucaena, Prosopis and Chamaecrista in West Africa (Senegal), East Africa (Sudan) and South America (Brazil). The type strain of Mesorhizobium plurifarium ORS 1032 has been deposited in the LMG collection as LMG 11892. PMID- 9731276 TI - Enterococcus asini sp. nov. isolated from the caecum of donkeys (Equus asinus). AB - Several Gram-positive, non-spore-forming and non-motile bacteria consisting of pairs or chains of cocci were isolated during an investigation of the bacterial flora of the caecum of donkeys. Physiological and metabolic data indicated that the strains belong to the genus Enterococcus; phenotypic traits of these organisms were not consistent with any of the currently known Enterococcus species. 16S rRNA gene sequence analysis placed these strains in the genus Enterococcus. Their closest relatives are Enterococcus avium, Enterococcus faecium and Enterococcus pseudoavium with a sequence similarity of 97.4%. This group of strains can be differentiated from the other Enterococcus spp. by their phenotypic characteristics: strains do not grow in 6.5% NaCl; they do not produce acid from mannitol, sorbitol, sorbose, sucrose, raffinose, ribose and tagatose; they produce acid from D-xylose; they are able to utilize pyruvate; and they present a negative reaction on arginine. The name Enterococcus asini sp. nov. is proposed for these strains; the type strain is AS2T (= DSM 11492T). PMID- 9731277 TI - An approach to characterizing uncultivated prokaryotes: the Grey Lung agent and proposal of a Candidatus taxon for the organism, 'Candidatus Mycoplasma ravipulmonis'. AB - An approach to characterizing uncultivated bacteria which combines a PFGE procedure for obtaining purified full-length chromosomes with PCR amplification is described. Isolated chromosomes from uncultivated organisms provide a specifically identifiable source material for hybridization, amplification and cloning. The availability of purified chromosomes for DNA amplification should aid in examining the diversity of microbial populations and should also reduce the possibility of forming hybrid DNA artifact molecules. The approach is illustrated by isolating the chromosome of the uncultivated agent of rodent Grey Lung disease and using the purified chromosomes to amplify and directly sequence the evolutionarily conserved 16S rRNA gene. The Grey Lung agent (GLA) contains a 650 kb chromosome and is shown to be a Mycoplasma sp. located phylogenetically in the hominis group of mycoplasmas. If a simple genomic lesion(s) is responsible for the unculturability of GLA, it is conceivable that complementation with DNA from a close relative could permit growth on artificial media. PMID- 9731278 TI - Taxonomic significance of 2,4-diaminobutyric acid isomers in the cell wall peptidoglycan of actinomycetes and reclassification of Clavibacter toxicus as Rathayibacter toxicus comb. nov. AB - An HPLC procedure which separates D- and L-amino acid isomers was applied to an analysis of peptidoglycan of 2,4-diaminobutyric acid (DAB)-containing actinomycetes. The cell wall peptidoglycans of species of the genera Agromyces, Clavibacter and Rathayibacter contain DAB and have been differentiated principally by their menaquinone profile. These peptidoglycans are known to be identical in structure, all being of the B2 gamma type, possessing both D- and L DAB. The type strains of all the subspecies of Clavibacter michiganesis have D- and L-DAB in almost equal proportions in their cell wall peptidoglycan as previously reported. In contrast, the type strains of Clavibacter toxicus and all valid species of the genera Agromyces and Rathayibacter contain the L-isomer of DAB almost exclusively. This characteristic is in good agreement with phylogenetic analyses based on 16S rDNA sequences and menaquinone profiles. On the basis of these data, the transfer of Clavibacter toxicus to the genus Rathayibacter as Rathayibacter toxicus comb. nov. is proposed. The isomer profile of DAB is shown to be a good taxonomic marker to differentiate these genera. PMID- 9731279 TI - Reclassification of Thermomonospora and Microtetraspora. AB - Almost complete 16S rRNA sequences from seven Thermomonospora strains, Thermomonospora curvata, Thermomonospora formosensis, Thermomonospora fusca, Thermomonospora mesophila, Thermomonospora chromogena, Thermomonospora alba and Thermomonospora mesouviformis (a synonym of Thermomonospora alba) were determined and subjected to phylogenetic analysis together with the sequences from all the representative members of the suborder Streptosporangineae. On the basis of phylogenetic, chemotaxonomic and phenotypic evidence, the transfer is proposed of Thermomonospora formosensis to the genus Actinomadura as Actinomadura formosensis comb. nov., Thermomonospora mesophila to the genus Microbispora as Microbispora mesophila comb. nov., and Thermomonospora fusca and Thermomonospora alba to a new genus, Thermobifida gen. nov., which belongs to the family Nocardiopsaceae, as Thermobifida fusca comb. nov. and Thermobifida alba comb. nov. Thermobifida alba is designated the type species of the genus. The transfer is also proposed of all species of the Microtetraspora pusilla group, which were transferred from Actinomadura, to a new genus, Nonomuria gen. nov., as Nonomuria africana comb. nov., Nonomuria angiospora comb. nov., Nonomuria fastidiosa comb. nov., Nonomuria ferruginea comb. nov., Nonomuria flexuosa comb. nov., Nonomuria helvata comb. nov., Nonomuria polychroma comb. nov., Nonomuria pusilla comb. nov., Nonomuria recticatena comb. nov., Nonomuria roseola comb. nov., Nonomuria roseoviolacea comb. nov., Nonomuria rubra comb. nov., Nonomuria salmonea comb. nov., Nonomuria spiralis comb. nov. and Nonomuria turkmeniaca comb. nov. Nonomuria pusilla is designated the type species of the genus. PMID- 9731280 TI - Description of Arthrobacter creatinolyticus sp. nov., isolated from human urine. AB - Three strains of creatinine-hydrolysing bacteria isolated from human urine were characterized taxonomically. They were aerobic, non-spore-forming, Gram-positive rods with the peptidoglycan of the cell wall containing lysine. MK-8 and MK-9 were found to be the major types of menaquinone. The G + C content of the DNA was 66-67 mol%. The 16S rRNA sequence of one strain (GIFU 12498) was determined and aligned with other high-G + C-content Gram-positive rods from different genera. Following phylogenetic analysis, this strain was placed in the genus Arthrobacter. Arthrobacter protophormiae was the most closely related species in the phylogenetic tree, and this species also showed the highest sequence homology value (97%) with GIFU 12498. However, DNA-DNA hybridization indicated that GIFU 12498 did not belong to A. protophormiae (33.8 +/- 3.5% chromosomal similarity). The three urine strains belonged to one species because they shared more than 95% DNA-DNA similarity. It is proposed that these strains are placed in the genus Arthrobacter as a new species, Arthrobacter creatinolyticus sp. nov. The type strain of A. creatinolyticus is GIFU 12498, which has been deposited in the Japan Collection of Microorganisms (JCM) with the accession number JCM 10102. PMID- 9731281 TI - Pelistega europaea gen. nov., sp. nov., a bacterium associated with respiratory disease in pigeons: taxonomic structure and phylogenetic allocation. AB - Twenty-four strains isolated mainly from infected respiratory tracts of pigeons were characterized by an integrated genotypic and phenotypic approach. An extensive biochemical examination using conventional tests and several API microtest systems indicated that all isolates formed a phenotypically homogeneous taxon with a DNA G + C content between 42 and 43 mol%. Whole-cell protein and fatty acid analysis revealed an unexpected heterogeneity which was confirmed by DNA-DNA hybridizations. Four main genotypic sub-groups (genomovars) were delineated. 16S rDNA sequence analysis of a representative strain indicated that this taxon belongs to the beta-subclass of the Proteobacteria with Taylorella equigenitalis as its closest neighbour (about 94.8% similarity). A comparison of phenotypic and genotypic characteristics of both taxa suggested that the pigeon isolates represented a novel genus for which the name Pelistega is proposed. In the absence of differential phenotypic characteristics between the genomovars, it was preferred to include all of the isolates into a single species, Pelistega europaea, and strain LMG 10982 was selected as the type strain. The latter strain belongs to fatty acid cluster I and protein electrophoretic sub-group 1, which comprise 13 and 5 isolates, respectively. It is not unlikely that the name P. europaea will be restricted in the future to organisms belonging to fatty acid cluster I, or even to protein electrophoretic sub-group 1, upon discovery of differential diagnostic features. PMID- 9731282 TI - Pseudonocardia asaccharolytica sp. nov. and Pseudonocardia sulfidoxydans sp. nov., two new dimethyl disulfide-degrading actinomycetes and emended description of the genus Pseudonocardia. AB - Seven bacterial strains capable of oxidizing methyl sulfides were isolated from experimental biofilters filled with tree-bark compost. The isolates could be divided into two groups according to their method of methyl sulfide degradation. Four isolates could use only dimethyl disulfide as the sole source of energy and three strains were able to use dimethyl sulfide and dimethyl disulfide. Oxidation of the methyl sulfides by both groups led to the stoichiometric formation of sulfate. Chemotaxonomic, morphological, physiological and phylogenetic properties identified all isolates as members of the genus Pseudonocardia. The absence of phosphatidylcholine from the polar lipid pattern, as well as results of 16S rDNA analyses, led to the proposal of two new species, Pseudonocardia asaccharolytica sp. nov. and Pseudonocardia sulfidoxydans sp. nov. The type strains are P. asaccharolytica DSM 44247T and P. sulfidoxydans DSM 44248T. With respect to the characteristic polar lipid pattern and the ability to oxidize sulfides, an emended description of the genus Pseudonocardia is proposed. PMID- 9731284 TI - Spiroplasma turonicum sp. nov. from Haematopota horse flies (Diptera: Tabanidae) in France. AB - Strain Tab4cT, a helical prokaryote that was isolated from the body of a Haematopota sp. fly collected in Champchevrier, Indre-et-Loire, Touraine, France, was found to be a member of the class Mollicutes. The cells of strain Tab4cT were small, motile helices that were devoid of a cell wall. The organism passed through filters with mean pore diameters as small as 0.20 mm. Strain Tab4cT grew rapidly in liquid SP-4 medium at both 30 and 37 degrees C. The organism fermented glucose but did not hydrolyse arginine or urea, and did not require serum for growth. In preliminary electrophoretic analyses, the cell protein patterns of strain Tab4cT were distinct from those of 14 other spiroplasmas found in mosquitoes, deer flies and horse flies from Europe and the Far-East. In reciprocal metabolism inhibition and deformation serological tests, employing antigens and antisera representative of spiroplasma groups I-XXXIII (including all sub-groups), plus ungrouped strains BARC 1901 and BARC 2649, no serological relationship with Tab4cT was found. The G + C content of the DNA of strain Tab4cT was about 25 +/- 1 mol% and its genome size was 1.305 kbp. It is proposed that spiroplasma strain Tab4cT be assigned to group XVII (presently vacant) and that strain (ATCC 700271T) is the type strain of a new species, Spiroplasma turonicum. PMID- 9731285 TI - Corynebacterium camporealensis sp. nov., associated with subclinical mastitis in sheep. AB - Four strains of a hitherto-unknown catalase-positive, facultatively anaerobic Corynebacterium species were isolated from the milk of sheep affected by subclinical mastitis. The most characteristic phenotypic reactions of the four strains were their weak fermentative acid production from glucose, their failure to produce acid from mannitol, xylose, sucrose and maltose, and a strong CAMP reaction with Staphylococcus aureus. Chemotaxonomic investigations revealed the presence of a cell wall based on meso-diaminopimelic acid and short-chain mycolic acids, which is consistent with the genus Corynebacterium. A comparative 16S rRNA gene sequence analysis confirmed that the organisms from sheep were members of the genus Corynebacterium, where they formed a distinct subline, exhibiting > 4% sequence divergence with other known Corynebacterium species. Based on both phenotypic and phylogenetic findings, a new species, Corynebacterium camporealensis, is proposed. The type strain of Corynebacterium camporealensis is CECT 4897 (= CRS-51T). PMID- 9731283 TI - Sulfurisphaera ohwakuensis gen. nov., sp. nov., a novel extremely thermophilic acidophile of the order Sulfolobales. AB - Three spherical thermoacidophilic archaea (strains TA-1T, TA-13, TA-14) were obtained from acidic hot springs located in Ohwaku Valley, Hakone, Japan. All the isolates are facultatively anaerobic, and grew optimally at around 85 degrees C, pH 2.0. Isolate TA-1T was characterized further. The G + C content of DNA from TA 1T is 33 mol%. Although these properties resemble those of the genus Acidianus, the sequence of the 16S rRNA gene from strain TA-1T was more similar to that of species of Stygiolobus than of Acidianus. DNA-DNA hybridization experiments also indicated that strain TA-1T is clearly distinguished phylogenetically from the members of Acidianus, Sulfolobus and Metallosphaera. On the basis of the distinct physiological and molecular properties, we describe the new strains as members of the new genus Sulfurisphaera. The type species of the genus is Sulfurisphaera ohwakuensis, and the type strain of the species is TA-1T (= IFO 15161T). PMID- 9731286 TI - Desulfobulbus rhabdoformis sp. nov., a sulfate reducer from a water-oil separation system. AB - A mesophilic, Gram-negative, rod-shaped, marine, propionate-oxidizing sulfate reducer (strain M16T) was isolated from a water-oil separation system on a North Sea oil platform. The optimum conditions for growth were 31 degrees C, pH 6.8-7.2 and 1.5-2.0% (w/v) NaCl and 0.1-0.3% (w/v) MgCl2.6H2O in the medium. The growth yield with sulfate was 4.6 g cell biomass (mol propionate oxidized)-1. Strain M16T is nutritionally related to members of the genus Desulfobulbus, but differs in that it has no vitamin requirement and is able to utilize fumarate and malate as carbon and energy sources. Hydrogenase activity measured as hydrogen uptake was mainly membrane-bound and varied with the growth substrate. Highest activity [28 mumol min-1 (mg protein)-1] was found in cells grown with hydrogen and lowest [50 nmol min-1 (mg protein)-1] in cells grown with propionate as electron donors for sulfate reduction. Desulforubidin, menaquinone-5(H2) and cytochromes of the c and b-type were present. The fatty acid pattern was similar to that found for Desulfobulbus propionicus. The DNA base composition was 50.6 mol% G + C. Strain M16T is equidistantly related to D. propionicus and Desulfobulbus elongatus with 96.1% 16S rDNA similarity. On the basis of differences in genotypic, phenotypic and immunological characteristics, strain M16T (= DSM 8777T) is proposed as the type strain of a new species, Desulfobulbus rhabdoformis. PMID- 9731287 TI - Desulfurella kamchatkensis sp. nov. and desulfurella propionica sp. nov., new sulfur-respiring thermophilic bacteria from Kamchatka thermal environments. AB - Two strains of moderately thermophilic bacteria, which reduce elemental sulfur to hydrogen sulfide, were isolated from volcanic sources in Kamchatka. Strain K-119T was obtained from a thermophilic microbial community associated with Thermothrix thiopara, and strain U-8T was isolated from a cyanobacterial mat inhabiting a sulfide-rich hot spring. Cells of both strains are short Gram-negative rods, motile with one polar flagellum (strain K-119T) or non-motile (strain U-8T). Both strains are obligate anaerobes, have temperature otima of 54-55 degrees C and pH optima of 6.9-7.2. Molecular hydrogen, acetate, fumarate, malate, pyruvate, lactate and long-chain saturated fatty acids served as growth substrates for both species; strain U-8T was also able to grow on propionate. All substrates were oxidized completely, H2S and CO2 being the only metabolic products. Elemental sulfur was obligately required for growth of strain K-119T, whereas strain U-8T was able to grow also with thiosulfate as electron acceptor and on pyruvate without an external electron acceptor. The DNA G + C contents of strains K-119T and U-8T were 31.6 and 32.2 mol%, respectively. Phenotypic features and the results of 16S rRNA sequencing indicate the affiliation of the new isolates to the genus Desulfurella. The DNA-DNA hybridization with Desulfurella acetivorans was 40% for strain K-119T and 55% for strain U-8T; the DNA-DNA hybridization between the new isolates was 32%. Based on the results of morphological, physiological and phylogenetic studies the following two new species are proposed: Desulfurella kamchatkensis sp. nov. with the type strain K-119T (= DSM 10409T) and Desulfurella propionica sp. nov. with the type strain U-8T (= DSM 10410T). PMID- 9731288 TI - Vibrio pectenicida sp. nov., a pathogen of scallop (Pecten maximus) larvae. AB - Five strains were isolated from moribund scallop (Pecten maximus) larvae over 5 years (1990-1995) during outbreaks of disease in a hatchery (Argenton, Brittany, France). Their pathogenic activity on scallop larvae was previously demonstrated by experimental exposure. The phenotypic and genotypic features of the strains were identical. The G + C content of the strains was in the range 39-41 mol%. DNA DNA hybridization showed a minimum of 73% intragroup relatedness. Phylogenetic analysis of small-subunit rRNA sequences confirmed that these strains should be affiliated within the family Vibrionaceae and that they are closely related to Vibrio tapetis and Vibrio splendidus. Phenotypic and genotypic analyses revealed that the isolates were distinct from these two vibrios and so constitute a new species in the genus Vibrio. They utilized only a limited number of organic substrates as sole carbon sources, including betaine and rhamnose, but did not utilize glucose and fructose. In addition, their responses were negative for indole, acetoin, decarboxylase and dihydrolase production. The name Vibrio pectenicida is proposed for the new species; strain A365 is the type strain (= CIP 105190T). PMID- 9731289 TI - Corynebacterium thomssenii sp. nov., a Corynebacterium with N-acetyl-beta glucosaminidase activity from human clinical specimens. AB - A strain of a previously undescribed non-lipophilic coryneform bacterium was isolated from pleural fluids of a patient with chronic renal failure, stroke and pneumonia. Slow fermentative acid production from glucose, maltose and sucrose, and strong N-acetyl-beta-glucosaminidase activity were the most characteristic features of the bacterium. Chemotaxonomic characterization unambiguously indicated that the organism belonged to the genus Corynebacterium. The results of comparative 16S rRNA gene sequence analysis revealed that the isolate represented a new species within the genus, for which the name Corynebacterium thomssenii sp. nov. is proposed. The type strain is DSM 44276. PMID- 9731290 TI - Streptomyces turgidiscabies sp. nov. AB - A new bacterial species is described, for which the name Streptomyces turgidiscabies is proposed. This organism causes potato (Solanum tuberosum) scab in eastern Hokkaido, Japan; the lesions caused are distinctly erumpent. In culture, S. turgidiscabies is distinct from other scab-causing Streptomyces species, having flexuous spore chains and grey mass colour. The spores of this organism are cylindrical and smooth. Its cell walls contain the LL-diaminopimelic acid isomer, and its DNA G + C content is 71 mol%. S. turgidiscabies does not produce melanin or other diffusible pigments, does not grow on agar media at pH 4.0 or 37 degrees C, is positive for utilization of raffinose and inulin as a carbon source, and is sensitive to streptomycin (20 micrograms ml-1), penicillin G (10 IU ml-1), polymyxin B (15 micrograms ml-1), and thallium acetate (10 micrograms ml-1). The levels of DNA relatedness within S. turgidiscabies strains are high but relatedness between strains of this species and strains of S. acidiscabies, S. scabies, S. caviscabies, S. griseus, S. setonii and S. tendae are low. The type strain is SY9113T (= ATCC 700248T = IFO 16080T). PMID- 9731291 TI - Characterization of Actinomyces turicensis and Actinomyces radingae strains from human clinical samples. AB - Whole-organism protein electrophoresis was used to compare and group unidentified coryneform bacteria resembling Gardnerella vaginalis and various Actinomyces and Arcanobacterium species. The obtained clusters of strains were further characterized by whole-cell fatty acid analysis and a variety of biochemical tests. Species-specific oligonucleotide probes based on 16S rRNA gene sequences were designed. The results demonstrate that the majority of the isolates belonged to Actinomyces turicensis; the other strains belonged to Actinomyces radingae. The descriptions of both species are emended. PMID- 9731292 TI - Staphylococcus succinus sp. nov., isolated from Dominican amber. AB - Two bacterial isolates, designated AMG-D1T and AMG-D2, were recovered from 25-35 million-year-old Dominican amber. AMG-D1T and AMG-D2 biochemically most closely resemble Staphylococcus xylosus; they differ physiologically from other staphylococci. Fatty acid analysis and comparisons with extensive databases were unable to show relatedness to any specific taxon. Moreover, AMG-D1T and AMG-D2 contain tuberculostearic acid and meso-diaminopimelic acid, characteristic of the G + C-rich coryneform bacteria, as opposed to L-lysine characteristic of staphylococci. AMG-D1T and AMG-D2 have a G + C ratio of 35 mol%. Phylogenetic analysis with the 16S rRNA gene indicated that AMG-D1T and AMG-D2 were most closely related to Staphylococcus equorum, S. xylosus, Staphylococcus saprophyticus and other novobiocin-resistant staphylococci. Stringent DNA-DNA hybridization studies with AMG-D1T revealed similarities of 38% with S. equorum, 23% with S. xylosus and 6% with S. saprophyticus. The results indicate that AMG D1T and AMG-D2 represent a novel species, which was named Staphylococcus succinus sp. nov. The type strain of the new species is AMG-D1 (ATCC 700337). PMID- 9731293 TI - Identification of streptococci from Greek Kasseri cheese and description of Streptococcus macedonicus sp. nov. AB - Taxonomic studies were performed on some Streptococcus-like organisms isolated from naturally fermented Greek Kasseri cheese. By SDS-PAGE analysis of whole-cell proteins the group was found to be quite different from Streptococcus thermophilus. Comparative 16S and 23S rRNA sequence analyses showed that the isolates represent a new species within the genus Streptococcus, where they are most closely related to the Streptococcus bovis cluster. On the basis of these phylogenetic results and some phenotypic differences, a new species, Streptococcus macedonicus, is proposed. The type strain is ACA-DC 206. PMID- 9731294 TI - Identification of bacterial isolates from biofilters as Paracoccus alkenifer sp. nov. and Paracoccus solventivorans with emended description of Paracoccus solventivorans. AB - Two groups of strains isolated from biofilters for the treatment of waste gases were assigned to the genus Paracoccus by phylogenetic and chemotaxonomic methods. All type strains of the genus Paracoccus were compared with these groups using 16S rDNA sequence analysis, fatty acid patterns and physiological reaction profiles. For both groups, the nearest related reference species was Paracoccus solventivorans based on 16S rDNA sequence similarity. However, whereas one group of isolates was identified as a member of this species by fatty acid analysis and DNA-DNA hybridization, the other group was proposed as a new species, Paracoccus alkenifer sp. nov. Fatty acid analysis showed the unusual fatty acid 20:1cis13 instead of 19:0 cyclo11-12 for P. alkenifer and P. solventivorans, and 14:1cis7 instead of 12:1cis5 for P. alkenifer and Paracoccus kocurii. By means of a GC-MS method, diaminopimelic acid was detected for P. solventivorans. Based on these results we propose an emended description for the species P. solventivorans. PMID- 9731295 TI - Roseobacter gallaeciensis sp. nov., a new marine bacterium isolated from rearings and collectors of the scallop Pecten maximus. AB - Four bacterial strains were isolated from larval cultures and collectors of the scallop Pecten maximus. They showed a high level of intragroup genomic relatedness (84-95%) as determined by DNA-DNA hybridization. The cells were Gram negative, strictly aerobic, motile, ovoid rods. They grew at temperatures from 15 to 37 degrees C and from pH 7.0 to 10, but did not grow in the absence of NaCl and required growth factors. They had the ability to use a wide variety of compounds as sole carbon source: D-mannose, D-galactose, D-fructose, D-glucose, D xylose, melibiose, trehalose, maltose, cellobiose, sucrose, mesoerythritol, D mannitol, glycerol, D-sorbitol, meso-inositol, succinate, propionate, butyrate, gamma-aminobutyrate, DL-hydroxybutyrate, 2-ketoglutarate, pyruvate, fumarate, glycine, L-alpha-alanine, beta-alanine, L-glutamate, L-arginine, L-lysine, L ornithine and L-proline. They exhibited oxidase and catalase activities but no denitrification activity. The isolates did not contain bacteriochlorophyll a. The G + C content ranged from 57.6 to 58 mol%. Phylogenetic analyses of the 16S rRNA sequence revealed that these isolates belong to the genus Roseobacter. On the basis of quantitative hybridization data, it is proposed that these isolates should be placed in a new species, Roseobacter gallaeciensis. The type strain is Roseobacter gallaeciensis BS107T (= CIP 105210T). PMID- 9731296 TI - Paracoccus marcusii sp. nov., an orange gram-negative coccus. AB - Phenotypic, chemotaxonomic and 16S rDNA sequence analysis of an orange Gram negative coccus that appeared as a contaminant on a nutrient agar plate delineated a new species of the genus Paracoccus. Phenotypic features of the strain that differ from all or most of the previously described Paracoccus species include its bright orange colour, caused by the synthesis of large amounts of carotenoids (mainly astaxanthin), and its inability to use nitrate as an electron acceptor in respiration. The name Paracoccus marcusii is proposed for this organism. The type strain is DSM 11574T. PMID- 9731297 TI - Burkholderia graminis sp. nov., a rhizospheric Burkholderia species, and reassessment of [Pseudomonas] phenazinium, [Pseudomonas] pyrrocinia and [Pseudomonas] glathei as Burkholderia. AB - In a survey of soil and wheat or maize rhizoplane bacteria isolated using a medium containing azelaic acid and tryptamine as sole carbon and nitrogen sources, respectively, a large proportion of Burkholderia-like bacteria were found. Among them, a homogeneous group of strains was identifiable based on phenotypic properties, fatty acid composition, DNA-DNA hybridizations and 16S rDNA sequences. According to molecular data, this group belongs to the genus Burkholderia but its weak similarity to previously described species suggests that it belongs to a novel species. Closest 16S rDNA phylogenetic neighbours of this species are Burkholderia caryophylli and two previously named Pseudomonas species which clearly appear to be part of the Burkholderia genus and were thus named Burkholderia glathei comb. nov. and Burkholderia phenazinium comb. nov. Strains of the new species are oxidase- and catalase-positive, produce indole and gelatinase, and use L-xylose, lactose, rhamnose, trehalose, D-lyxose, L-arabitol, xylitol and D-raffinose as sole carbon source. This novel taxon is named Burkholderia graminis. In the course of this study, [Pseudomonas] pyrrocinia also proved to be a member of the Burkholderia genus. PMID- 9731298 TI - Bacillus horti sp. nov., a new gram-negative alkaliphilic bacillus. AB - Novel Gram-negative alkaliphilic strains were isolated from soil obtained from Atsuma, Hokkaido, Japan. The isolates were strictly aerobic rods that produced subterminally located ellipsoidal spores. Chemotaxonomic characteristics of the isolates included the presence of meso-diaminopimelic acid in the cell wall and a DNA G + C content of 40.2-40.9 mol%. The major isoprenoid quinone was menaquinone 7 and the cellular fatty acid profile consisted of a significant amount of 15-C branched-chain acids, iso-C15:0 and anteiso-C15:0. The growth rate was higher at pH 8-10 than at pH 7. Comparative sequence analysis of 16S rDNA of 14 alkaliphilic Bacillus strains indicates that the isolated strain has an equidistant relationship to three already defined rRNA groups of alkaliphilic Bacillus species. Based on the morphological and physiological characteristics, as well as phylogenetic position as determined by 16S rDNA analysis and DNA-DNA relatedness data, it is concluded that these isolates should be designated as a new species, for which the name Bacillus horti is proposed. The type strain is K13T (= JCM 9943T). PMID- 9731299 TI - Vibrio halioticoli sp. nov., a non-motile alginolytic marine bacterium isolated from the gut of the abalone Haliotis discus hannai. AB - Six alginolytic, facultatively anaerobic, non-motile marine bacteria were isolated from the gut of abalone Haliotis discus hannai. DNA-DNA hybridization data showed that the six strains constituted a single genospecies. Phylogenetic analyses of 16S rDNA sequences indicated that the isolates should be assigned to the genus Vibrio. The phenotypic features of the isolates were closely related to Vibrio fischeri and Vibrio pelagius biovar I, but 13 traits (motility, luminescence, alginase production, lipase production, lysine decarboxylase, indole production, growth in 1 and 6% NaCl and assimilation of five carbon compounds) distinguished these strains from V. fischeri, and 17 traits (motility, growth at 37 degrees C, lipase production, indole production, growth in 1 and 6% NaCl, acid from sucrose and D-sorbitol, and assimilation of nine carbon compounds) distinguished these strains from V. pelagius. The G + C content of the isolates was 41.6-43.1 mol%. According to DNA-DNA hybridization data and 16S rDNA phylogenetic analyses, it was concluded that the six isolates constitute a new species different from any other Vibrio species. The name Vibrio halioticoli sp. nov. (type strain IAM 14596T) is proposed. A set of phenotypic features which enables differentiation of the new species from other species of the Vibrionaceae family is described. PMID- 9731300 TI - Phylogenetic diversity of Streptococcus suis strains of various serotypes as revealed by 16S rRNA gene sequence comparison. AB - The 16S rRNA gene sequences of reference strains of Streptococcus suis serotypes 1-34 and 1/2 were determined. A comparative sequence analysis showed that the degree of sequence similarity between S. suis reference strains ranged from 93.94 to 100%. A dendrogram was constructed from the similarity matrix. Thirty-two strains representing 32 serotypes fell into a major group divided into three clusters. The other strains, S. suis serotypes 32, 33 and 34, were more distant. Biochemical characterization of the six more distant strains, including S. suis serotypes 20, 22, 26, 32, 33 and 34, revealed a profile similar to that of other S. suis serotypes. Comparison of the 16S rRNA gene sequences of S. suis reference strains with sequences of other members of the genus Streptococcus indicated that, with the exception of S. suis serotypes 32, 33 and 34, reference strains did not cluster with any other species in the genus. In conclusion, 16S rRNA gene sequence analysis defined a major group of S. suis reference strains which were very closely related and a higher divergence for S. suis serotypes 32, 33 and 34. However, to date, there is no strong evidence to reclassify strains of these serotypes in another species. PMID- 9731301 TI - Phylogenetic heterogeneity of the genus Williopsis as revealed by 18S rRNA gene sequences. AB - A phylogenetic investigation of the ascomycetous yeast genus Williopsis was performed by using 18S rRNA gene sequence analysis. Comparative sequence analysis revealed the genus to be phylogenetically heterogeneous. The five varieties of Williopsis saturnus [var. mrakii, var. sargentensis, var. saturnus (type), var. suaveolens and var. subsufficiens] were found to have identical 18S rRNA gene sequences and formed a distinct group, quite separate from all other Williopsis and non-Williopsis species examined. Williopsis mucosa was found to be the closet phylogenetic relative to the Williopsis saturnus group, however a sequence divergence of approximately 2.3% suggests this species may belong to a separate genus. The recently described species Williopsis salicorniae was found to exhibit a relatively close association with Ogataea minuta (identical to Pichia minuta), the type species of the genus Ogataea. The remaining two members of the genus, Williopsis californica and Williopsis pratensis, were found to form distinct lineages, displaying no specific association with any other Williopsis or non Williopsis species. Based on comparative analysis of 18S rRNA genes it is apparent that the genus Williopsis as presently constituted is not monophyletic, and that the five currently recognized species form separate sublines each potentially worthy of separate generic status. The genus Williopsis should be restricted to the type species Williopsis saturnus and its five varieties. Despite the five varieties of Williopsis saturnus being genealogically indistinguishable at the 18S rRNA gene level, sequence analysis of the Internal transcribed spacer (ITS) region revealed that the five varieties could be differentiated on both their ITS1 and their ITS2 sequences, providing further evidence of the value of ITS sequences for discrimination of yeasts at the subspecies level. PMID- 9731302 TI - Comparative ribosomal protein (L11 and L30) sequence analyses of several Streptomyces spp. commonly used in genetic studies. AB - The taxonomic relationships among nine strains of Streptomyces, which have been commonly used for genetic studies, were examined by sequence analysis of their ribosomal L11(= rplK) protein genes. Phylogenetic relationships among these organisms derived from similarity sequence analysis of the rplK genes were in good agreement with those derived from the analysis of the deduced L11 protein amino acid sequence itself, indicating complete sequence homology among Streptomyces coelicolor A3(2), 'Streptomyces lividans 66' and Streptomyces violaceoruber JCM 4423. S. coelicolor A3(2) related (in the order of closer relatedness) to Streptomyces antibioticus ATCC 14888, Streptomyces griseus IFO 13189, Streptomyces lavendulae MA 406 A-1 and Streptomyces virginiae MAFF 6014. Sequence analysis of the 26 N-terminal amino acid residues of ribosomal L30 proteins also resulted in similar phylogenetic relationships, except that S. griseus, S. lavendulae and S. virginiae were not differentiated from each other using this method. These findings concerning the phylogenetic relationship therefore confirm the previous conclusion that S. coelicolor A3(2), 'S. lividans 66' and S. violaceoruber should be recognized as a single taxon at the species level. PMID- 9731303 TI - Corynebacterium phocae sp. nov., isolated from the common seal (Phoca vitulina). AB - Phenotypic and phylogenetic studies were performed on four strains of a Gram positive non-acid-fast coryneform-like organism isolated from the nasal cavities of common seals (Phoca vitulina). Chemotaxonomic investigations revealed the presence of corynomycolic acids in the unidentified isolates, which is consistent with corynebacteria. 16S rRNA gene sequence analysis demonstrated that the strains from seals represent a hitherto unknown subline within the genus Corynebacterium sensu stricto. Based on the results of the phylogenetic analysis and phenotypical criteria, it is proposed that the bacterium should be classified as a new species, Corynebacterium phocae. The type strain of Corynebacterium phocae is CCUG 38205T. PMID- 9731304 TI - Phylogenetic relationships of Salmonella based on rRNA sequences. AB - To establish the phylogenetic relationships between the subspecies of Salmonella enterica (official name Salmonella choleraesuis), Salmonella bongori and related members of Enterobacteriaceae, sequence comparison of rRNA was performed by maximum-likelihood analysis. The two Salmonella species were separated by 16S rRNA analysis and found to be closely related to the Escherichia coli and Shigella complex by both 16S and 23S rRNA analyses. The diphasic serotypes S. enterica subspp. I and VI were separated from the monophasic serotypes subspp. IIIa and IV, including S. bongori, by 23S rRNA sequence comparison. PMID- 9731305 TI - Staphylococcus pulvereri = Staphylococcus vitulus? PMID- 9731306 TI - Adsorption and leaching of trifluralin, metolachlor, and metribuzin in a commerce soil. AB - Trifluralin [2,6-dinitro-N,N-dipropyl-4-(trifluoromethyl)benzenamine], metolachlor [2-chloro-N-ethyl-6-methylphenyl)-N-(2-methoxy-1-methylethyl) aceta mide], and metribuzin [4-amino-6-(1,1-dimethylethyl)-3-(methylthio)-1,2,4, triazin-5(4H) -one] were selected to study adsorption and leaching potentials related to pollution on Commerce silty clay loam soil near Baton Rouge, Louisiana. At a I:10 soil/water ratio, the Koc values for trifluralin, metolachlor and metribuzin were 875, 135, and 96, respectively. Leaching of these herbicides was evaluated in soil columns (5.4 cm i.d. x 26 cm long). Total recoveries of the herbicides applied to the soil column were 73.1% +/- 4.1%. When the soil columns were leached with three pore volumes of water, the distributions of trifluralin in soil and leachate were 99.993% and 0.007% of the total recoveries, respectively. The distributions of metolachlor was 65.27% in soil and 34.7% in leachate. The distributions of metribuzin was 11.42% in soil and 88.58% in leachate. The results showed that metolachlor and metribuzin were readily leached, while trifluralin was strongly adsorbed to soil. Leaching of three herbicides in the soil column followed the leaching trends of their calculated leaching indices 1.41 x 10(4), 4.18 x 10(6), and 3.38 x 10(8) for trifluralin, metolachlor, and metribuzin, respectively. The results of the study demonstrated the potential of pollution for metolachlor and metribuzin to be leached into the ground water in soils with shallow aquifer. PMID- 9731307 TI - Herbicide residues in leaves of Erythroxylum coca var. coca plants treated with soil-applied tebuthiuron and hexazinone. AB - The herbicide residue levels in leaves of Erythroxylum coca var. coca Lam. plants treated with soil applications of tebuthiuron and hexazinone at 3.36 and 6.72 kg a.i. ha-1 were determined in order to estimate the potential for human exposure to these residues from consuming the leaves or cocaine produced from them. Field grown plants were treated with a commercial formulation of tebuthiuron or hexazinone and leaves were harvested at the first indication of herbicide injury (i.e. chlorosis and/or necrosis) and at the onset of leaf abscission. Herbicide residues were detected by HPLC in leaf samples from both harvests of all plants treated with tebuthiuron or hexazinone. At 3.36 kg ha-1, herbicide residues in the leaves were less than 2 micrograms g-1 dry wt. for both harvests of both experiments. The highest residue levels detected were 5.90 micrograms g-1 dry wt. for tebuthiuron and 7.17 micrograms g-1 dry wt. for hexazinone in leaves from plants treated with the herbicide at the rate of 6.72 kg ha-1 and harvested at the onset of leaf drop. Based on published toxicity data and estimates of leaf consumption, the herbicide residues in leaves of E. coca var. coca plants treated with tebuthiuron or hexazinone at twice the recommended control rates or less would have a negligible contribution to the health risks of individuals who chew coca leaves. Furthermore, based on the most conservative estimates of cocaine yield and herbicide carry over, death by cocaine overdose would occur long before the NOEL for either herbicide was reached. PMID- 9731308 TI - Pesticide concentration variations correlated with well bore volume removal in shallow coastal plain ground water. AB - The effects of well bore volume removal (Vn) on the concentration of alachlor [2 chloro-N-(2,6-diethylphenyl)-N-(methoxy methyl) acetamide] and prometon (6 methoxy-N,N'-bis(1-methylethyl)-1,3,5-triazine-2,4-diamine] in ground water obtained from three monitoring wells installed in the Coastal Plain region of North Carolina was investigated. Seasonal effects were also investigated by conducting the exercise in February and May. In the majority of cases, the lowest pesticide concentrations occurred in the initial well bore volume (V1 = stagnant water). Removal of additional well bore volumes (V2 to V10) from two of the wells resulted in pesticide concentrations that did not vary substantially. This indicates that a representative aquifer sample was obtainable, in most cases from these wells, after removal of the initial well bore volume. In contrast, a third well required the purging of two well bore volumes before a stable alachlor concentration was achieved. Seasonal effects of bore volume removal vs. pesticide concentrations for the three wells were not significant (P > 0.05). It was concluded that a protocol for improved accuracy in pesticide analyses of ground water can be obtained by establishing a pesticide concentration-purging (well bore volume) relationship for each well. PMID- 9731309 TI - Fractions of calcium in the plant-soil system affected by the application of olive oil wastewater. AB - A pot experiment using calcareous soil was conducted in a growth chamber to examine the effects of olive oil wastewater applications on Ca fractions in the plant and on exchangeable Ca in soil after plant growth. The experimental treatments consisted of two rates of olive oil wastewater, two mineral fertilizer treatments including K, which supplied K in amounts equivalent to the K supplied by the olive oil wastewater treatments, a mineral fertilizer without K treatment (F), and a control. The pots were sown with ryegrass which was harvested 3 times at monthly intervals. The high rate of olive oil wastewater resulted in a prolonged increases in dry matter production. It also resulted in a reduction in the concentrations of soluble Ca, bound Ca, inorganic insoluble Ca and organic insoluble Ca in the plant shoots relative to the control and the F treatment. The low rate of olive oil wastewater produced similar but less marked effects, with decreases being observed in the soluble Ca and bound Ca fractions. After 3 months of plant growth, soil exchangeable Ca was higher in the control and F treatment soils than in the soils receiving olive oil wastewater or K fertilizer. At this time, there were no significant differences in exchangeable Ca between the soils receiving olive oil wastewater and those treated with K fertilizer. PMID- 9731310 TI - All that you can be: stereotyping of self and others in a military context. AB - The authors tested the shifting standards model (M. Biernat, M. Manis, & T. E. Nelson, 1991) as it applies to sex- and race-based stereotyping of self and others in the military. U. S. Army officers attending a leadership training course made judgments of their own and their groupmates' leadership competence at 3 time points over a 9-week period. We examined the effects of officer sex and race on both subjective (rating) and objective/common-rule (ranking/Q-sort) evaluations. Stereotyping generally increased with time, and in accordance with the shifting standards model, pro-male judgment bias was more evident in rankings than in ratings, particularly for White targets. Self-judgments were also affected by sex-based shifting standards, particularly in workgroups containing a single ("solo") woman. Differential standard use on the basis of race was less apparent, a finding attributed to the Army's explicit invocation against the use of differential race-based standards. PMID- 9731311 TI - On being happy and mistaken: mood effects on the fundamental attribution error. AB - Does temporary mood influence the occurrence of the fundamental attribution error (FAE)? Based on recent affect-cognition theorizing and research on attributions, 3 experiments predicted and found that negative moods decrease and positive moods increase the FAE, because of the information-processing consequences of these affective states. In Experiment 1, happy mood enhanced and sad mood reduced dispositional attributions based on coerced essays advocating unpopular opinions. Experiment 2 replicated this effect using an unobtrusive mood induction in a field study. Experiment 3 further confirmed these results and also showed that changes in the FAE were linked to mood-induced differences in processing style, as indicated by memory data and confirmed by mediational analyses. The results are discussed in terms of the cognitive processing strategies that mediate mood effects on attributions. The implications of the findings for everyday inferences and for contemporary theories of affect and cognition are considered. PMID- 9731312 TI - The illusion of transparency: biased assessments of others' ability to read one's emotional states. AB - Three sets of studies provide evidence for an illusion of transparency, or a tendency for people to overestimate the extent to which others can discern their internal states. People often mistakenly believe that their internal states "leak out" more than they really do. The authors attribute this bias to a tendency for people to adjust insufficiently from the "anchor" of their own phenomenological experience when attempting to take another's perspective. Evidence for this illusion is provided by showing that liars overestimate the detectability of their lies (Studies 1a, 1b, and 1c) and that people believe their feelings of disgust are more apparent than they actually are (Studies 2a and 2b). A final pair of experiments (Studies 3a and 3b) explores the implications of the illusion of transparency for people's reluctance to intervene in emergencies. All 3 sets of studies also provide evidence consistent with the proposed anchoring and adjustment interpretation. PMID- 9731313 TI - Partner verification: restoring shattered images of our intimates. AB - When spouses received feedback that disconfirmed their impressions of their partners, they attempted to undermine that feedback during subsequent interactions with these partners. Such partner verification activities occurred whether partners construed the feedback as overly favorable or overly unfavorable. Furthermore, because spouses tended to see their partners as their partners saw themselves, their efforts to restore their impressions of partners often worked hand-in-hand with partners' efforts to verify their own views. Finally, support for self-verification theory emerged in that participants were more intimate with spouses who verified their self-views, whether their self views happened to be positive or negative. PMID- 9731314 TI - Membership has its (epistemic) rewards: need for closure effects on in-group bias. AB - Three studies examined the impact of the need for cognitive closure on manifestations of in-group bias. All 3 studies found that high (vs. low) need for closure increased in-group favoritism and outgroup derogation. Specifically, Study 1 found a positive relation between need for cognitive closure and both participants' ethnic group identification and their collective self-esteem. Studies 2 and 3 found a positive relation between need for closure and participants' identification with an in-group member and their acceptance of an in-group member's beliefs and attitudes. Studies 2 and 3 also found a negative relation between need for closure and participants' identification with an out group member and their acceptance of an out-group member's beliefs and attitudes. The implications of these findings for the epistemic function of in-groups are discussed. PMID- 9731315 TI - The moderating effect of self-esteem in reaction to voice: converging evidence from five studies. AB - It has been posited that high self-esteem persons (high SEs) are more confident than low self-esteem persons (low SEs) of their capability to provide meaningful input in a decision process. If this is so, then high SEs should be more influenced by their perceived level of voice, relative to low SEs. Survey data from 4 field studies showed that voice was more positively related to various dependent variables among high SEs than low SEs. In Study 5, the authors experimentally manipulated voice as well as participants' beliefs about their capability to provide meaningful input. As expected, voice had a greater impact on the reactions of participants who were led to believe that they were more capable of providing meaningful input. Theoretical implications are discussed. PMID- 9731316 TI - Culture, social organization, and patterns of violence. AB - Traditional social theorizing holds that strong and cohesive family, community, and religious institutions rein in violence. However, in cultures where certain types of violence are condoned, this should not be true. Specifically, in the U.S. South and West, where culture-of-honor traditions persist, greater social organization should be associated with more violence. This pattern was confirmed in examinations of argument-related homicide rates (Study 1); mass consumption patterns for violence in entertainment, recreation, and vocational pursuits (Study 2); and voting patterns of political elites on gun control and national defense issues (Study 3). Across the 3 studies, social organization was associated with effects in the South and West opposite of what they were in the North. Implications for general theories of cultural evolution, suggesting a cycle in the way societies crystallize and change, are discussed. PMID- 9731317 TI - Adult attachment style and affect regulation: strategic variations in self appraisals. AB - Four studies examined the link between adult attachment style and strategic variations in self-appraisals. Whereas secure persons held a stable positive self view, Studies 1-2 showed that avoidant persons showed a positive self-view and anxious-ambivalent persons a negative self-view, which were strengthened by distress arousal and weakened by factors that inhibit the activation of regulatory mechanisms. Studies 3-4 indicated that insecure persons' self-views varied in accordance with specific attachment-related concerns and needs. Avoidant persons' positive self-view was related to their attempts to validate their sense of self-reliance, and anxious-ambivalent persons' negative self-view was related to their attempts to win others' compassion and affection. Results are discussed in terms of attachment-related strategies of affect regulation. PMID- 9731318 TI - Adult attachment style and affect regulation: strategic variations in subjective self-other similarity. AB - Six studies examined the link between adult attachment style and subjective self other similarity. In Studies 1-3, data were collected on representations of self other similarity in the realms of traits and opinions. Studies 4-5 examined the effects of affective inductions on the link between attachment and self-other similarity. Study 6 examined the cognitive maneuvers people differing in attachment style use for changing self-other similarity upon distress arousal. Whereas avoidant persons underestimated self-other similarity and anxious ambivalent persons overestimated it, secure persons provided more accurate similarity scores. These differences were exacerbated by negative affect and mitigated by positive affect. Insecure persons' distortions resulted from transformations they made in representations of the self and others. Results are discussed in terms of attachment theory. PMID- 9731319 TI - Moderators of self-other agreement: reconsidering temporal stability in personality. AB - Accurate prediction requires information not only about central tendencies but also about variability. In personality prediction, however, most research has focused on trait-level central tendencies. Previously proposed moderators of personality prediction are all conceptually similar in comparing an individual's central tendency in response patterns with that of the normative person. This article proposes an alternative: Trait-level prediction is enhanced by measuring the temporal stability of response patterns within persons. Across 2 studies, individuals with temporally stable response patterns had higher self-other agreement on conscientiousness and extraversion than did individuals with less temporally stable patterns. By comparison, normatively based variables (interitem variability, scalability, or construct similarity) did not moderate self-other agreement. The implications for personality structure, assessment, and prediction are discussed. PMID- 9731320 TI - The dynamic self: how the content and structure of the self-concept change with mood. AB - Change in the content and structure of the self-concept was examined in a 2-year prospective study of depression. A self-descriptive card-sorting task measured the self-concepts of participants who were either high or low in cognitive vulnerability to depression at 2 times, once when their mood was low and once when it was not. Analyses examined change in the positive and negative content of the self-concept and in 3 features of self-structure: differential importance, compartmentalization, and self-complexity. Findings suggest that change in the content of the self-concept simply reflects life circumstances, whereas change in self-structure may help to counteract stress and negative mood. PMID- 9731321 TI - Personal goals and emotional well-being: the moderating role of motive dispositions. AB - Two studies examined the importance of motive dispositions in determining the extent to which the pursuit of personal goals accounts for interindividual differences in emotional well-being. Within the domains of agency and communion, motives were assessed with a picture-story test, whereas self-report measures were used to assess goal attributes. Study 1 found that progress toward motive congruent goals, in contrast to progress toward motive-incongruent goals, accounted for students' daily experiences of emotional well-being. Study 2 found that the combination of high commitment to and high attainability of motive congruent goals predicted an increase in students' emotional well-being over 1 semester. In contrast, high commitment to motive-incongruent goals predicted a decline in emotional well-being. Results are discussed with reference to a 2 system approach to human motivation. PMID- 9731322 TI - Top-down, bottom-up, and horizontal models: the direction of causality in multidimensional, hierarchical self-concept models. AB - A new structural equation modeling approach to questions of the direction of causal flow between global and specific multidimensional measures of self-concept (SC) in two 2-wave, longitudinal studies demonstrated that (a) higher order factors were unable to explain relations among first-order factors at Time 1 (T1), at Time 2 (T2), or between T1 and T2; (b) T1 global SC had little effect on specific SC factors at T2 (a top-down model), but specific factors at T1 had even less effect on T2 global SC (a bottom-up model); and (c) many specific factors were more stable than global factors, but higher order factors were most stable. Results provide little support for top-down, bottom-up, or reciprocal models, instead arguing for a horizontal model in which each T2 SC factor is primarily a function of the matching T1 SC. This casts further doubt on the usefulness of hierarchical representations of SC. PMID- 9731323 TI - Reactions to penalties for offenses committed by the police and public citizens: testing a social-cognitive process model of retributive justice. AB - This research investigated situations involving the police and public citizens in which both committed offenses and were punished. Participants responded to either a scenario describing police violence against a "green" protester (n = 177) or one describing dangerous driving by detectives leading to injury of juveniles in a car chase (n = 149). Results showed that information about following orders mitigated participants' reactions to offenses committed by authority figures. The perceived seriousness of the offenses committed by police authorities was negatively related to participants' level of right-wing authoritarianism and positively related to the importance participants assigned to universalism values. These relations were opposite in direction for the public offenders who refused to obey a police order. Other relations supported a social-cognitive process model in which values, responsibility, seriousness, and deservingness are key variables. PMID- 9731324 TI - The self-fulfilling prophecy in close relationships: rejection sensitivity and rejection by romantic partners. AB - The authors hypothesized a self-fulfilling prophecy wherein rejection expectancies lead people to behave in ways that elicit rejection from their dating partners. The hypothesis was tested in 2 studies of conflict in couples: (a) a longitudinal field study where couples provided daily-diary reports and (b) a lab study involving behavioral observations. Results from the field study showed that high rejection-sensitive (HRS) people's relationships were more likely to break up than those of low rejection-sensitive (LRS) people. Conflict processes that contribute to relationship erosion were revealed for HRS women but not for HRS men. Following naturally occurring relationship conflicts, HRS women's partners were more rejecting than were LRS women's partners. The lab study showed that HRS women's negative behavior during conflictual discussions helped explain their partners' more rejecting postconflict responses. PMID- 9731325 TI - Making sense of loss and benefiting from the experience: two construals of meaning. AB - Theoretical models of the adjustment process following loss and trauma have emphasized the critical role that finding meaning plays. Yet evidence in support of these models is meager, and definitions of meaning have been too broad to facilitate a clear understanding of the psychological process involved. Using a prospective and longitudinal study of people coping with the loss of a family member, we differentiate 2 construals of meaning--making sense of the event and finding benefit in the experience--and demonstrate that both independently play roles in the adjustment process following the loss. Results indicate that making sense of the loss is associated with less distress, but only in the 1st year postloss, whereas reports of benefit finding are most strongly associated with adjustment at interviews 13 and 18 months postloss. PMID- 9731326 TI - Wavelet transforms for electrocardiogram processing. AB - Wavelet transforms were used to remove noise and detect characteristic points for feline and human electrocardiograms. The electrocardiograms contained both 60 Hz line noise and wideband noise. Noise was removed from the signals using both Fourier and wavelet techniques. For the Fourier technique, the signals were notch filtered at 60 Hz, and then low-pass filtered. To remove noise using wavelet techniques, the signals were wavelet-transformed and then soft thresholding was performed on each of the resulting details. The two methods of removing noise were then compared to illustrate the advantages of the wavelet approach. Wavelet transforms were also used to detect a characteristic point on each beat. An algorithm was written to find this point, calculate the instantaneous heart rate, and then plot heart rate variability. PMID- 9731327 TI - A World Wide Web accessible multi-species ECG database. AB - We have developed a system for remotely accessible secure electronic storage of electrocardiographic (ECG) and other associated data. It allows entry of data from any authorized remote user and is specifically built to accommodate the ECGs of multiple species. The present system is implemented on a Sun Sparc Solaris 2.5 platform using Oracle 7.3.2, and the Oracle 7.3.2 Web server. It may be easily ported to any other UNIX or Windows NT platform. No client is needed other than an Internet Protocol connected computer using a web browser such as Netscape Navigator or Microsoft Internet Explorer. PMID- 9731328 TI - Simulation of cardiac conduction system in distributed computer environment. AB - A high resolution, three dimensional, computer model of the cardiac conduction system has been developed. Cardiac geometry was constructed from sectional images of VHP project of the National Library of Medicine. The heart was modeled as a matrix of cells that fill its anatomical structure. The intracellular distance was 1 mm and the total number of cells were 457,482. Electrophysiological parameters like action potential, absolute refractory period and conduction velocity were assigned to each of the cells. The pattern of the excitation sequence propagation as well as potentials on the body surface points were computed on a single processor. The working memory and the time for computation of the algorithms were minimized using efficient data structures. The time to compute an excitation sequence over one cardiac cycle was 4 hours. The algorithms were also implemented on a distributed network of personal computers running on a QNX operating system. The speed of computation of the excitation sequence algorithm was improved by a factor of 2.52 when the algorithm was implemented on a network of three Intel-66 MHz machines. PMID- 9731329 TI - The realization of a haptic (force feedback) interface device for the purpose of angioplasty surgery simulation. AB - Simulation is becoming an increasingly accepted method for training personnel for complex or high risk tasks. With the advent of modern Virtual Reality (VR) technology, such simulations are becoming more and more immersive. One significant limitation to the advance of this science is the challenges associated with simulating the touch or feel of various tasks in conjunction with VR simulations. This work has focused on creating and assessing the feasibility of haptic technology for high fidelity surgery simulation. It has culminated in a prototypical interface device to provide tactile feedback during the simulation of complex interventional radiology procedures such as Angioplasty. This device extends state of the art motion control, distributed computing processes, high resolution sensors for real-time feedback, and modular design techniques to accomplish these goals. The realization of the device, its capabilities, interfacing requirements, its limitations, and future extensions will be presented here. PMID- 9731330 TI - Programmed stimulation for simulation of atrial tachyarrythmias. AB - Swine have become the preeminent non-human animal model for testing of cardiac assistance devices. Such devices must be robust to perturbations in the EKG; therefore it is expeditious, as part of cardiac assistance device testing, to simulate arrhythmias such as atrial flutter and fibrillation in swine. Methods for producing atrial tachyarrythmias include surgical ablation or crushing of atrial tissue and the administration of drugs. These are dangerous, unreliable and usually irreversible; in varying combinations and degrees. When we applied rapid evenly spaced electrical atrial pacing in swine, the variability of ventricular response was less than expected from the corresponding arrhythmia in man. We will describe efforts to produce reversible atrial tachyarrythmias in swine, using programmed stimulation, with the desired degree of ventricular response variability. PMID- 9731331 TI - A DSP based real time system for analysis of bundle of His and late potentials using wavelet transforms. AB - A new system for analysis of Bundle of HIS and Late Potentials has been developed using the wavelet approach. The objective of the present research work is to develop a real-time system which does not rely on averaged data and has the capability to detect beat to beat variations in the cardiac micro-volt signals from the body surface recordings. Multiresolution wavelet analysis gives better time and frequency resolution of the signal and its implementation on DSP hardware makes the system real time. The clinical applicability of the system developed is currently being investigated with initial success on pre clinical data. PMID- 9731332 TI - Feature extraction and modeling of the variability of performance in terms of biomechanical motion patterns during MMH tasks. AB - In investigating manual material handling (MMH) jobs, such as lifting, the quantification of the various kinematic and kinetic parameters of the lift is an important step towards functional assessment and evaluation. Experimental data collection generates a large quantity of data for the different kinetic, kinematic, and electromyographic parameters over the various lifting cycles. In order to efficiently manage and interpret the data, it is important to use appropriate tools which would reduce the dimension of the original data set without sacrificing any important features. Furthermore, the generated parameters are often expressed as a function of the lifting cycle resulting in complex waveforms as the unit of analysis. Appropriate statistical analysis of these waveforms or motion profiles should reflect their vectorial constitution as a function of the lifting cycle rather than the usual method of using traditional descriptive statistics based on collapsing the data over the cycle. PMID- 9731333 TI - Using CAI to accommodate a variety of learning styles in a biomechanics course. AB - Multimedia technology offers a more interactive approach to instruction than the traditional classroom lectures. Through computer-aided instruction (CAI), a number of teaching styles can be used that take into account the different preferences of the students. The Biomechanics Tutorial program that the authors have written is a CAI that incorporates audio, video, simulations, and graphics to: review concepts of mechanics (kinematics and kinetics of interconnected rigid bodies), familiarize students with functional anatomy, and allow students to interactively evaluate the law of mechanics applied to physical performance of activities modeled by a set of biomechanical models of the joints. Principles of ergonomics are reinforced by enabling the student to perform numerous numerical experiments within the context of workplace or task redesign and see the real time consequences of these alterations. For example, the task of holding a load is simulated by allowing the student to change elbow and shoulder angles and the orientation and magnitude of the load. The consequences of these in terms of required muscle forces and joint reaction forces at the elbow and shoulder will be updated on the screen. The detailed rationale of developing this Biomechanics Tutorial which integrates a variety of learning styles will be presented. PMID- 9731334 TI - A technique for using strain gauges to evaluate airbag interaction with cadaveric upper extremities. AB - Radius and ulna fractures from airbag deployment onto the forearm have been reported in the literature. Based on laboratory experiments with eight cadaveric upper extremities, this paper presents a method for using strain gages to evaluate upper extremity loading during airbag deployment. The technique provides strain rates, bending moments, and time of fracture for the radius and ulna. Planar rosettes (350 omega, 5% strain) were selected as the best choice given the application to bone with a rosette being placed mid-shaft on both the anterior and posterior surfaces of the radius and ulna. Forearm incisions were intended to be minimally invasive and to limit damage to the interosseous membrane. The bone surface was prepared with Ether, and the gauges were bonded to the surface with methyl-2-cyanoacrylate. A thin latex cover was installed over the surface of the rosettes to isolate the gauges from the surrounding tissue. Strain relief of the gauges was provided by securing the wire leads to the bone with tie-wraps, as well as suturing the wires to the skin. With this technique all gauges reported accurate data throughout the duration of the impact. PMID- 9731335 TI - The mechanics of drop landing on a flat surface--a preliminary study. AB - In an effort to investigate the mechanics of drop landing on a flat surface, bilateral performance characteristics during the landing were investigated. Two force plates were used as landing surfaces, and the six component kinetic time histories and the path of the center of pressure were measured for both feet. The forces were combined to obtain the resultant ground reaction force, allowing comparison between the global resultant and the forces developed by each leg. The results highlighted an asymmetry, and the presence of one dominant leg during the landing: the contact with the plate occurred with a delay between the dominant and non-dominant legs, and the forces applied to the plate were also unequal. Furthermore, due to the high rate of data acquisition and sensitivity of the force plates used, it was possible to correlate the peaks present in the global vertical reaction force with the impact of the different anatomical segments of each foot. The availability of such high resolution information presents to researchers and clinicians with more key identifiers to quantify various phenomena, potentially injury prevention, pathological diagnosis or quantification, and others. PMID- 9731336 TI - An experimental study of airbag impact to the orbit using an instrumented Hybrid III headform. AB - This paper provides the results from an experimental study intended to assess airbag impact to the orbit. Twenty-seven airbags were deployed onto an instrumented Hybrid III headform. Seven different types of airbags were used that included tethered and nontethered, light to heavy nylon weaves, and different coating types. The airbags were deployed via a pneumatic deployment system. Seven individual force transducers, each having a 2.25 cm2 contact area, were placed on the right orbital region to evaluate the force patterns. The midpoint of the two ocular regions of the headform was positioned 12.5 cm above and 15.0 cm away from the center of the airbag, a position determined from previous airbag deployments to yield the highest leading edge velocity. The average maximum force per sensor ranged from 15.4 N to 63.6 N, and peak pressure ranged from 68 kPa to 282 kPa. The upper center of the orbit presented the highest values while the center of the orbit recorded the lowest values, a comparison that was proven to be statistically significant. For this configuration, the maximum force on the ocular region was found to be independent of the presence of a tether or the maximum internal airbag pressure. PMID- 9731337 TI - Anticipatory postural adjustment and T'ai Chi Ch'uan. AB - Recently much attention has focused on T'ai Chi Ch'uan as a method of improving balance and reducing the risk of falls in the elderly. Little has been done to determine the mechanism of these improvements. This study examined the effects of a sixteen week training program on anticipatory postural adjustments (APA). Eight subjects (median age 36.5) in good health, with no prior training in T'ai Chi were tested. The battery of tests included load dropping of 2.2 kg loads by the subjects standing on an unstable board placed on a force platform. The level and direction of instability were varied. The results show counterintuitive reductions in the APAs of several muscle groups while the stability of standing improved. We interpret the findings as an indication that practicing T'ai Chi leads to a greater use of the elasticity of the peripheral structures involving muscles, ligaments, and tendons while the participation of the central neutral structures in postural equilibrium is decreased. PMID- 9731338 TI - Peak detection in auditory and somatosensory evoked potentials by means of the zero-crossings wavelet representation. AB - A feature detection algorithm for detecting singularity-like features in a one dimensional signal has been developed. It is based on the zero-crossings wavelet representation introduced by Mallat [1], and involves a variance-weighted cross correlation of the appropriate wavelet transform of the signal in the various frequency bands with an averaged template characterizing the particular feature. The algorithm was developed for the purpose of automatic feature detection in averaged evoked potentials. It successfully detects waves I to V in the auditory evoked potential and the N20 and P25 peaks in the median nerve stimulated somatosensory evoked potential, in normal and delayed-latency cases. The performance of the algorithm is plotted as a function of the signal-to-noise ratio. A priori knowledge of the expected feature amplitudes and latencies is not necessary, but may be incorporated into the model. By feeding the feature amplitude and latency estimates back into the algorithm as a priori knowledge, it is configured to track the changing characteristics of a feature over time. PMID- 9731339 TI - Development of an integrated automated retinal surgical laser system. AB - Diabetes is a serious disease that affects millions of people world wide. One of the many complications that may result from this disease is an eye disorder called diabetic retinopathy. This side effect is characterized by the leaching of fluid, mainly blood, into the retinal tissue ultimately killing the rods and cones in the area. Scientists have discovered that by occulating and destroying some of the capillaries near the infected area, further damage may be limited and further vision loss prevented. This procedure is performed on the retina by using a Continuous Wave argon laser to place a pattern of lesions that kill the rods, cones, and underlying capillaries. The "Computer Aided Laser Optics System for Ophthalmic Surgery" or CALOSOS is a system that fully integrates the placement of therapeutic retinal lesions through computer control. To date, two different sub systems have been used to achieve the desired accuracy needed including lesion placement and depth; an all-digital system and a hybrid analog/digital system. The current tracking system that precisely controls the laser uses a wide-angle lens camera to view the whole retina. The difficulty lies in the fact that the only way to control the depth of the lesion is to measure the central reflectance. The goal of this research is to establish that there is a linear relationship between the reflectance of the lesion and the depth of the lesion when this wide-angle lens is used. PMID- 9731340 TI - Wavelet analysis of electromyography for back muscle fatigue detection during dynamic constant-torque exertions. AB - The fatigue of the back muscles appears to be strongly implicated as a risk factor for acquisition of low back pain, which is one of the leading ills of our industrial society. Previously, researchers have successfully measured the level of muscular fatigue by using the Fourier transform to analyze the frequency content of the electromyogram (EMG). However, due to the requirement that the EMG signal be stationary, the Fourier transform is suitable only for the analysis of static muscle exertions in which the muscle is held at constant length and tension. Because the majority of industrial work tasks are not static in nature, new methods for quantifying fatigue during dynamic work are needed. The wavelet transform is a novel, although mathematically well developed, technique for analyzing non-stationary signals that has only recently been applied to the study of EMG. Consequently, the main objective of this project is to develop techniques, using the wavelet transform, for the quantification of back muscle fatigue during dynamic repetitive working conditions. PMID- 9731341 TI - A new ambulatory monitoring instrument of posture and mobility related activities. AB - Long-term ambulatory monitoring of posture and mobility related activities provides useful information about the extent of disability and the outcome of rehabilitation. The aim of this project is developing an instrument which could distinguish between a set of selected mobility-related physical activities and produce parameters characterising the subjects' activity pattern; The selected activities are lying, sitting, standing and walking. A novel activity transducer and a data logger with 1MB memory are employed in the system configuration. Analytical algorithms and program are developed. The subject's whole day activity pattern and the histogram of each activity event are successfully obtained. PMID- 9731342 TI - Use of a turbine in a breath-by-breath computer-based respiratory measurement system. AB - The Computer-Based Respiratory Measurement System (CBRMS) is capable of analyzing individual breaths to monitor the kinetics of oxygen uptake, carbon dioxide production, tidal volumes, pulmonary ventilation, and other respiratory parameters during rest, exercise, and recovery. Respiratory gas volumes are measured by a calibrated turbine transducer while the respiratory gas concentrations are measured by a calibrated, fast-responding medical gas analyzer. To improve accuracy of the results, the inspiratory volumes and gas concentrations are measured and not assumed to be equal to expiratory volumes or ambient concentrations respectively. The respiratory gas volumes and concentration signals are digitized and stored in arrays. The gas volumes are converted to flow signals by software differentiation. These digitized data arrays are stored as files in a personal computer. Time alignment of the flow and gas concentration signals is performed at each breath for maximum accuracy in analysis. For system verification, data were obtained under resting conditions and under constant load exercises at 50 W, 100 W, and 150 W. These workloads were performed by a healthy, male subject on a bicycle ergometer. A strong correlation existed between the CBRMS steady-state results and the standard end-expirate bag collection technique. Thus, there is reason to believe that the CBRMS is capable of calculating respiratory transient responses accurately, a significant contribution to an understanding of total respiratory system function. PMID- 9731343 TI - Gender differences in muscular strength: an allometric model approach. AB - Despite the fact that allometric modeling has been shown to be a more valid technique for group comparisons of body-size-adjusted muscular strength, no study to date has compared gender differences using absolute (no adjustment for body mass, BM), ratio (force/BM) and allometric (force/BMa) models for an active duty military population. Our purpose in this study was to use these models to compare gender differences in three strength tasks: maximal weight lifted to 152 cm height, maximal isometric force on an upright cable pull and maximal isometric handgrip force. Subjects were 989 and 987 active duty men and women, respectively, each of whom had fat-free mass (FFM) assessed hydrostatically. Results indicate that the ranges of gender difference in strength (%) were 57.9 103.7, 14.5-64.7 and 18.7-77.9 for absolute, ratio and allometrically scaled expressions, respectively, for BM and FFM. We conclude: 1. Men exhibit greater strength than women for all three indices regardless of model approach. 2. The magnitude of these differences is smaller when considering FFM rather than BM, but is larger when considering allometric vs. ratio models. PMID- 9731344 TI - Airbag-induced eye injuries: experiments with in situ cadaver eyes. AB - In an attempt to investigate eye injuries from airbags, a set of experiments was performed that involved the deployment of several types of airbags onto thirteen unembalmed, previously frozen cadaver heads. The airbags differed in the material, coating, presence of a tether, and folding pattern, and were deployed via a pneumatic deployment system. The eyes were impacted in situ after being repressureized with saline injected through a 30-gauge needle. Injury determination was achieved by ophthalmic ultrasound imaging, staining with fluorescein dye, and dissection. All twenty-six eyes revealed detached retinas, as shown by the ultrasound, before impact as a result of decaying tissue and the freezing process. High speed video and film were used to capture the events. The impact velocities of the airbags were recorded from the digitized film at the first contact location with the eye and ranged from 30 m/s to 66 m/s. Eyeglasses were placed on four of the specimens, and the presence of eyeglasses seemed to provide protection to the eye because of the lack of contact between the airbag and ocular region. Minimal ocular damage was recorded for all experiments. PMID- 9731345 TI - Bilateral within-subject Q angle asymmetry in young adult females and males. AB - Previous investigations of the quadriceps angle (or Q angle) and its relationship to knee disorders have yielded equivocal results. Methodological differences may account for some of the observed discrepancies, but so too may the tendency for investigators to rely on between-group rather than within-subject research designs. The purpose of this investigation was to study the between-group (male versus female) and within-subject (right versus left lower limb) differences in Q angle measures in subjects with no history of knee disorders. The right and left Q angles of fifty young men and women were measured goniometrically with subjects standing in an erect, weight-bearing position. In males, there was little disparity in the magnitude of the Q angle between the right (9.5 degrees +/- 4.6) and left (10.4 degrees +/- 5.7) lower limb. Similarly, in females, the mean value for the right Q angle (10.5 degrees +/- 4.2) was only slightly smaller in magnitude than that of the left (12.2 degrees +/- 5.2). A 2 x 2 ANOVA revealed no significant gender (p < 0.17) or right to left lower limb (p < 0.19) differences. However, when the data for each subject was examined on an individual basis, the magnitude of the right and left Q angles differed by 4 degrees or more in 24 subjects. In 10 subjects, this difference ranged from 8.0 degrees to 10.3 degrees. For the majority of subjects the left Q angle was greater in magnitude than the right Q angle. These results suggest that (I) the assumption of symmetry in right versus left lower limb Q angle measures may be erroneous, (II) statistical comparisons of mean values may not be the best method by which to analyze Q angle data, and (III) further investigation of bilateral within-subject Q angle asymmetry is warranted. PMID- 9731346 TI - Enhancement of Xe-133 ventilation lung scan image acquired after Tc-99m perfusion scan. AB - In nuclear medicine, the match and mismatch between the images of lung perfusion and ventilation provide an important criterion to diagnose pulmonary embolism. Usually, for imaging clarity, the ventilation scan using 133Xe is performed before the perfusion scan using 99mTc. But the inverse order is preferred clinically, since (i) if the perfusion image is normal, there is no need to do the ventilation scan, and (ii) if the perfusion image is abnormal, the ventilation image can be obtained by focusing on the perfusion abnormalities. However, the quality of the ventilation image is reduced if the ventilation scan is performed after the perfusion scan, because the 140 keV photons emitted from 99mTc will scatter into the acquisition window of 133Xe (81 keV). The purpose of this study is to apply image processing techniques to reduce the scattering effect of 99mTc to get a better 133Xe ventilation image. First, an image sequence in the preferred inverse order is simulated using the images acquired in the normal order. Image processing techniques are used to find an optimized way to reduce the scattering background in the ventilation image. Second, a real image sequence is acquired in the inverse order. An improved ventilation image is then obtained by applying image processing techniques to this image sequence. PMID- 9731347 TI - In vitro characterization of a zinc based bioceramic. AB - Six different blends of zinc calcium phosphorous oxide ceramics (ZCAP) were prepared by mixing and calcining powders (ZnO:CaO:P2O5) of weight percent ratio: 50:30:20, 48:32:20, 44:20:26, 40:40:20, 30:40:30, and 30:30:40. ZCAP is a resorbable bioceramic and has been used to repair bone defects and deliver drugs in a continuous manner. The chemical composition, porosity, and elements released on exposure to buffered Tris HCl were measured for each blend of ZCAP. The products of mixing and thermal reaction were beta-Ca3(PO4)2, alpha-CaZn2(PO4)2, and 2CaO.P2O5. Free calcium and/or zinc oxide was present in several blends. The components of ZCAP and their volume percentages influenced the interconnected porosity of ZCAP bioceramics. The interconnected porosity for all blends of ZCAP ranged from 35 to 38%. Pore sizes from these six blends of ZCAP ranged from less 1 micron to greater than 100 microns. Results of the 12 hour dissolution study showed that more calcium was released than zinc or phosphorous from all blends of ZCAP. Zinc was released in trace amounts from all blends of ZCAP. Release of phosphorous from the different blends of ZCAP was not detected by the procedures used to detect phosphorous in this investigation. These blends of ZCAP have the potential to be used as bone substitutes and probably long term treatment of zinc deficiency in humans. PMID- 9731348 TI - A zinc based self setting ceramic bone substitute for local delivery of testosterone. AB - Testosterone has been shown to stimulate bone healing. However, large doses of testosterone are associated with liver damage and prostate enlargement. Continuous release of testosterone at the traumatized bone site could enhance healing without inducing systemic side effects. In this investigation a self setting zinc sulfate calcium phosphate (ZSCAP) ceramic with and without testosterone was used to fill experimentally induced bone defects in rats. Each treatment group consisted of six albino, Holtzman rats. Groups I and II consisted of non-operated and sham-operated animals. Rats in Group III, IV, V, and VI were implanted with ZSCAP particulate ceramic powders (63-75 microns) containing 0.0, 0.5, 1.0, or 1.5 mg testosterone in a 2.3 mm defect in the femur of each rat. Radiographic and morphologic examination of the implant site at four, eight, and twelve weeks post surgery showed integration of the implant in the femur of each rat. Serum testosterone of all rats was assayed at four, eight, and twelve weeks post surgery. Differences in the serum testosterone levels of rats in the six groups were not significant. Results of this study suggest that a ZSCAP testosterone composite can be used to repair traumatized bone without increasing the levels of testosterone in the systemic circulation. PMID- 9731349 TI - Controlled diffusional release of dispersed solute drugs from biodegradable implants of various geometries. AB - Chronic diseases and pathological medical conditions requiring the administration of longterm pharmaceutical dosages have in the past been treated by oral administrations of tablets, pills and capsules or through the use of creams and ointments, suppositories, aerosols, and injectables. Such forms of drug delivery, which are still currently used today, provide a prompt release of the drug, but with significant fluctuations in the drug levels within various regions of the body. Repeated administrations of the drug are often needed, at rather precise intervals of time, in order to maintain these levels within a relatively narrow therapeutic range as a means of assuring effectiveness at the low end and of minimizing adverse effects at the higher end of the fluctuation spectrum. Recent technical advances now permit one to control the rate of drug delivery. The required therapeutic levels may thus be maintained over long periods of months and years through implanted rate-controlled drug release capsules. Two such novel drug delivery systems currently employed are implanted erodible polymeric and ceramic capsules. Mathematical modeling and computer simulations can be very effective in improving and optimizing the performance of the self-regulating release of therapeutic drugs into specific regions of the body. Further development is needed for the optimal design of such capsules. It is in this area, in particular, that a review will be presented of the mathematical modeling techniques susceptible to refine the development of a reliable tool for designing and predicting the resulting pharmaceutical dosages as a function of time and space. Of primary importance in such models are the time-varying effective permeability of the capsule to the various molecules composing the drug, the effective solubility and diffusion coefficients of the drug and its metabolites in the surrounding tissues and fluids and, finally, the uptake of the drug at the target organ. Mathematical models are presented for the diffusional release of a solute from an erodible matrix in which the initial drug loading c0 is greater than the solubility limit cs. An inward moving diffusional front separates the reservoir (unextracted region) containing the undissolved drug from the partially extracted region. The mathematical formulation of such moving boundary problems has wide application to heat transfer with melting phase transitions and diffusion-controlled growth of particles, in addition to our topic of controlled release drug delivery. In spite of this diversity of applications, only a very few mathematical descriptions have been published for the analysis of release kinetics of a dispersed solute from polymeric or ceramic matrices. In these rare instances, perfect sink conditions are assumed, while matrix swelling, concentration-dependence of the solute diffusion coefficient and the external mass transfer resistance have been largely neglected. The ultimate goal of such an investigation is to provide a reliable design tool for the fabrication of specialized implantable capsule/drug combinations which will deliver pre specified and reproducible dosages over a wide spectrum of conditions and required durations of therapeutic treatment. Such a mathematical/computational tool can also prove effective in the prediction of suitable dosages for other drugs of differing chemical and molecular properties which have not been subjected to time-consuming animal laboratory testing. Finally, such models may permit more realistic scaling of the required dosages of therapeutic drug for variations in diverse factors such as body weight or organ size and capacity of the patient (clinical medicine) or animal (veterinary medicine for farm animals). Additional applications of controlled-release drug delivery for insecticide and pesticide use in agriculture, and the control of pollution in lakes, rivers, marshes, etc. in which a pre-programmed dose-time schedule is necessary, further PMID- 9731350 TI - A hydroxyapatite system for the continuous release of coumadin an anticoagulant. AB - Coumadin is an anticoagulant used in the treatment of patients with pulmonary thromboembolisms or those implanted with mechanical vascular devices. Its efficacy is complicated by side effects such as hemorrhaging. Administration of coumadin by the traditional routes of pills and injections causes wide fluctuations of the drug in plasma. Thus, experiments were conducted to develop a hydroxyapatite ceramic system for releasing coumadin in vivo continuously for 7 10 days. A series of preliminary in vivo experiments was conducted to determine an effective non-lethal therapeutic dosage of coumadin for rats. Homogenous matrices containing varying amounts of coumadin were implanted subcutaneously in rats and the blood clotting time was determined. Results of these experiments showed that one mg coumadin delivered from HA implants doubled the blood clotting time and did not induce fatal hemorrhage. Subsequent in vitro studies were conducted to determine the most efficient way to deliver coumadin at a slow and sustained rate. Results of these experiments suggested that a solid insert of one mg of compressed coumadin in a hydroxyapatite ceramic shell released the drug more slowly and efficiently than a homogenous matrix consisting of compressed coumadin and hydroxyapatite. PMID- 9731351 TI - In vitro and in vivo dapsone release from hydroxyapatite reservoirs. AB - Dapsone is a drug which is administered orally, on a daily basis, for four to five years as a cure for leprosy. The major problems associated with dapsone therapy include hemolysis, methemoglobinemia and patient non-compliance. Cimetidine reduces the side effects of dapsone and increases its levels in the blood. A ceramic drug delivery system was developed to maintain therapeutic levels of dapsone for an extended period of time and to alleviate associated side effects. In vitro release of commercial dapsone tablets (100 mg) and cimetidine pellets (400 mg) from hydroxyapatite reservoirs was studied in 100 ml absolute ethanol at 37 degrees C. Hydroxyapatite (HA) reservoirs loaded with one dapsone tablet released the drug at the rate of 8.3 mg/day for nine days, after which a much slower release occurred for another three days. With a load of two dapsone tablets, the rate of release was 6.7 mg/day for four weeks. HA reservoirs loaded with one cimetidine pellet delivered the drug at a rate of 25 mg/day for sixteen days. Combining both drugs in a single reservoir did not affect their respective release rates. When implanted subcutaneously in rats, HA reservoirs loaded with one dapsone tablet appeared to be well tolerated. After nine weeks, 77 mg of dapsone had been released. These experiments showed that HA reservoirs can be used to deliver dapsone and cimetidine in vivo. PMID- 9731352 TI - Cellular responses to various levels of sustained delivery of testosterone in the ventral prostate. AB - The objective of this study was to evaluate the effect of various dosages of testosterone (T) delivered in a sustained manner by means of tricalcium phosphate lysine (TCPL) delivery system on morphological changes of prostatic tissue using adult male rats as a model. In this experiment, adult male rats (250-300 g BW) were randomly divided into five equal groups (n = 8). Rats in group I, II, and III were castrated and implanted subcutaneously with TCPL loaded with three different dosages (10, 100 and 200 mg T, respectively) of T. Rats in group IV were castrated and implanted with sharm TCPL capsules, and rats in group V served as intact unimplanted controls. Surgical aseptic techniques were performed according to standard laboratory procedures. At the end of 4 and 12 weeks post implantation, four animals from each group were sacrificed and the prostate tissues were collected, weighted, and embedded for histo-pathological evaluations. Data collected from this study have shown that exogenous intake of T delivered in a sustained manner for twelve weeks induced several pathophysiological conditions in ventral prostatic tissue in comparison to the control and sham operated groups. This phenomenon was found to be directly proportional to the dose or the level of sustained delivery. The results demonstrated that the use of 10 mg filled TCPL implants decreased the total mass weight of ventral prostate. Light microscopic evaluation of this group (Group I) revealed a cellular adaptation through an atrophy in the epithelium component. Cytopathological observations such as low cuboidal and thin glands, pleomorphism, and occasional presence of connective tissue stroma were detected. In contrast, ventral prostate collected from animals implanted with TCPL filled with 200 mg T (Group III) showed a significant increase in weights of the wet prostatic tissues in comparison to all groups. Histopathological evaluations demonstrated the following. (i) prostatic hypertrophy alone, or in conjunction with hyperplasia of the epithelial cells, (ii) less connective tissue stroma in comparison to the control group, (iii) occasional involvement of mitotic figures, and (iv) increased angiogenesis. No significant change was observed in those animals implanted with TCPL capsules containing 100 mg T compared to the intact control animals. PMID- 9731353 TI - Germinal cell alterations associated with sustained delivery of testosterone enanthate in adult male rats. AB - It is well documented that testosterone stimulates protein anabolism, muscular development, bone growth and increased metabolic rate. In addition, exogenous low levels of testosterone can inhibit the secretion of gonadotropins by the anterior pituitary. The objective of this study was to investigate the effect of sustained delivery of testosterone enanthate (TE) on the testicular architecture by means of tricalcium phosphate-lysine(TCPL). In this experiment adult male Sprague Dawley rats (250-300 g BW) were randomly divided into three equal groups (n = 16). Rats in group I were implanted subcutaneously with TE loaded TCPL implants (4-6 ng/ml). Rats in group II were implanted with sham TCPL capsules, and rats in group III served as intact unimplanted controls. Surgical aseptic techniques were performed according to standard laboratory procedures. At the end of 2 and 4 weeks post implantation, eight animals from each group were sacrificed and the testes were collected, weighted, and embedded for histopathological evaluations. The results of this study revealed the following: (i) Sustained delivery of TE reduced spermatogonia numbers in animals after 2 and 4 weeks by approximately 10% in comparison to control animals. (ii) The lumen of the tubules showed slight regression after 4 weeks of TE treatment. (iii) Overall decrease of the vasculature of the experimental animals was seen after 2 and continued through 4 weeks with an occasional increase of microvasculature seen in animals treated for 4 weeks with TE. (iv) Finally, the germinal epithelium showed significant reduction in secondary spermatocytes and conversion to mature sperm. PMID- 9731354 TI - Biochemical surface modification of Ti-6Al-4V for the delivery of protein to the cell-biomaterial interface. AB - Biochemical surface modification involves delivery of biomolecules to the tissue implant interface to induce desired cell and tissue responses. We have previously had success in immobilizing and retaining bioactive molecules on Co-Cr-Mo but not on Ti-6Al-4V. The purpose of this study was to modify the gamma aminopropyltriethoxysilane (APS) scheme to enable successful attachment of protein to the surface of Ti-6Al-4V. Ti-6Al-4V samples were silanized with organic (acetone) solutions of APS and dried at increasing temperatures. Concentrations resulting in 2-4 NH2 per nm2 of nominal surface area were incubated in physiological saline for up to 96 hr to assess retention of amino groups. A model protein, trypsin, was coupled to silanized Ti-6Al-4V via glutaraldehyde. The samples were then incubated in saline, and the activity of residual immobilized enzyme was quantified. After drying at 45, 80, or 115 degrees C, the NH2 groups were lost from the surface by 24 hr of incubation in saline. On samples dried at 150 degrees C, with 4% APS, the number of NH2 groups increased after 8 hr and remained relatively constant through 96 hr. Likewise, at 150 degrees C with 2% APS the surface density of NH2 groups reached a maximum at 24 hr and remained relatively constant up to 96 hr. When incubated for 96 hr, Ti 6Al-4V with 4% APS and dried at 150 degrees C retained approximately 31% of the activity initially immobilized, whereas protein on 45 degrees C or adsorbed samples was lost by 24-48 hr. PMID- 9731355 TI - Inhibition of migration of endothelial cells by type V collagen and high molecular weight kininogens. AB - The failure of synthetic vascular grafts in humans is complex and related not only to the type of material but also to the accumulation of cellular and noncellular components onto or within the material. Proteins that adsorb on biomaterials such as vascular grafts may be involved in modulating cell function. Of these components, type V collagen and high molecular weight kininogens (HMWKs) are known to localize on the surface of vascular grafts. We have investigated the effect of type V collagen and HMWKs in the prevention of endothelialization of vascular prostheses by inhibiting cell migration. Inhibition of endothelial cell migration may lead to incomplete endothelialization and thus, failure of these devices. Various concentrations of type V collagen and HMWK solutions were placed at the bottom of 24 well tissue culture plates with porous membrane inserts (8 microns pore size) placed on top. Suspended human umbilical vein endothelial cells (HUVECs) were placed on top of the insert and incubated at 37 degrees C for 2 hours. Following incubation, the inserts were fixed and the cells that migrated through the pores were counted by microscopy. Our results showed that migration of endothelial cells was significantly inhibited in a dose dependent manner by type V collagen and HMWKs. PMID- 9731356 TI - Protein adsorption on chemically modified surfaces. AB - Following the implantation of a biomaterial, the first event to occur at the tissue-material interface is protein adsorption. Once proteins have adsorbed to the surface of a material, cells no longer see the material, but only the protein coated surface layer. This adsorbed protein layer can mediate the type of cells that adhere to the surface, which ultimately can determine the type of tissue that develops. In this experiment, glass and polystyrene surfaces were chemically modified forming ionic, non-ionic, hydrophobic, and hydrophilic surfaces. The modified surfaces were incubated in a RPMI diluted serum solution. After incubation, a radioimmunoassay was used to quantify the exposed proteins adsorbed onto the surface of the material. In general, albumin, complement C3 (C3), fibronectin (FN) and vitronectin (VN) competitively adsorbed to the modified surface in a similar fashion, whereas IgG adsorption was the opposite. The hydrophobic surfaces had higher adsorbance of the adhesion proteins (C3, FN and VN) compared to higher adsorption of albumin and IgG onto the hydrophilic surfaces. The surface mobility of the silane modified surfaces also affected the adhesion of proteins. The differences seen in protein adsorption did not directly correlate to monocyte adhesion and Foreign Body Giant Cell (FBGC) development on these surfaces. PMID- 9731357 TI - Computer analysis of human depth EEG in different sleep stages. AB - This study compares delta (< 4 Hz), theta (4-8 Hz), alpha (8-12 Hz) and beta (12 20 Hz) band EEG signals during wakefulness, rapid-eye-movement (REM) sleep, and stage 1 and stage 2 non-REM sleep recorded from both surface and depth electrodes in patients with drug-resistant partial seizures. Computer analysis utilizing Fast Fourier Transforms (FFT) was performed with the Neurovision software developed for this purpose. Mean amplitudes were calculated for each of the frequency bands. Preliminary analysis was performed with emphasis on the presence and characteristics of theta activity in the hippocampus of the brain. Results demonstrate theta wave activity in the hippocampus with an increase of theta activity in REM sleep as compared to non-REM sleep (stages 1 and 2). PMID- 9731358 TI - Predicting joint moments and angles from EMG signals. AB - Electromyographic (EMG) signals are a direct reflection of muscle activity. Efforts have been made to extract additional information from them regarding the kinematic results of muscle use, with limited success. The purpose of this study was to conduct a wheelchair propulsion experiment using a limited number of subjects to gather information for developing a neural network model to correlate EMG signals to kinematic features of the wrist, elbow, and shoulder joints. The trained model was used to predict joint dynamics from previously unseen EMG data. EMG signals were collected from four muscle groups while five able-bodied subjects propelled a wheelchair. The procedure was videotaped with the kinematics and dynamics of the three joints obtained by digitizing and transforming the images into numerical data. A neural network model, based on the back-propagation algorithm, was built and trained on the collected data. The net's predicted values were close to the observed values, particularly for the joint moments (within 7% of actual values). The results demonstrate the potential of neural networks for predicting the movement patterns of wheelchair users from a small number of subjects. PMID- 9731360 TI - Influence of frame rate and image delay on virtual driving performance. AB - The control and navigation of unmanned ground vehicles (UGVs) by humans requires a thorough understanding of the limitations in human perception and performance. Images of the external world recorded by cameras mounted on the UGV are presented as a video display to the operator, who then remotely manipulates the vehicle using a standard control. Operator performance is directly proportional to the computational complexity associated with the processing of video data. This work studies the effects of frame rate and image delay (lag) on remote driving performance. Experiments were conducted with five subjects using a driving simulator with a 1 dof force feedback steering wheel control. After sufficient training on the simulator, subjects drove a virtual car on a standard track under varying settings of frame rate and lag. Performance was measured by the duration to complete the course. Comparison of performance both within and between subjects showed characteristic driving patterns at different settings. Implications of the findings are discussed in relation to video data presentation for remote driving applications. PMID- 9731359 TI - Evaluation of FNS control systems: software development and sensor characterization. AB - Functional Neuromuscular Stimulation (FNS) systems activate paralyzed limbs by electrically stimulating motor neurons. These systems have been used to restore functions such as standing and stepping in people with thoracic level spinal cord injury. Research in our laboratory is directed at the design and evaluation of the control algorithms for generating posture and movement. This paper describes software developed for implementing FNS control systems and the characterization of a sensor system used to implement and evaluate controllers in the laboratory. In order to assess FNS control algorithms, we have developed a versatile software package using Lab VIEW (National Instruments, Corp). This package provides the ability to interface with sensor systems via serial port or A/D board, implement data processing and real-time control algorithms, and interface with neuromuscular stimulation devices. In our laboratory, we use the Flock of Birds (Ascension Technology Corp.) motion tracking sensor system to monitor limb segment position and orientation (6 degrees of freedom). Errors in the sensor system have been characterized and nonlinear polynomial models have been developed to account for these errors. With this compensation, the error in the distance measurement is reduced by 90 % so that the maximum error is less than 1 cm. PMID- 9731361 TI - The effect of restricting arm swing during normal locomotion. AB - This work examined possible perturbations in contra-lateral upper and lower limb motion, movement variability, and thoracic rotation, when a healthy mature adult walked a distance of 12 m at self-paced speed with and without unilateral passive arm restraint. The 2-dimensional walking data captured from 11 reflective markers placed on the subject's right side were recorded sagitally along the plane of progression using an electronic camera and Peak-5 Performance Video-system. The positional data were calculated off-line and differentiated with respect to one representative stride using Lab View software. Data were analyzed for 5 successive trials of normal walking; 5 successive walking trials with unilateral passive restraint; and a further 4 without restraint. Under the restrained condition the results showed: a) A significant decline in horizontal displacement of the limbs; b) higher variability of the trajectories generated, particularly at the ankle; c) a trend towards greater thoracic rotation on the unrestrained side; and d) altered angular velocity profiles for all upper and lower limb joints and trunk, which did not always return to baseline values after restraint removal. Since motivation level, environment, and methodology was consistent across trials, the findings suggest relatively stereotyped locomotor patterns in mature humans can be readily modulated in response to task constraints. PMID- 9731362 TI - Synergistic action by neuropeptide Y (NPY) and norepinephrine (NE) on food intake, metabolic rate, and brown adipose tissue (BAT) causes remarkable weight loss in the obese (fa/fa) Zucker rat. AB - NPY has been reported to co-exist within catecholaminergic neurons of the central and peripheral nervous systems. The functional significance in noradrenergic neurons has been related to the vasomotor effects of NPY which complement and interact with NE which is known to have central and peripheral effects on resting metabolic rate (RMR), food intake and body weight of rats. We have studied the effect of chronic peripheral administration of NPY on the metabolic action of NE in obese adult male rats. A group of 20 adult male obese (fa/fa) Zucker rats were acclimated to environmental temperature of either 28 degrees C or 17 degrees C. Each group was divided into 5 subgroups: (I) untreated controls; (II) Carrier treated Controls; (III) NPY treated; (IV) NE treated and (V) NPY + NE treated. In subgroups II-V, Alzet (2002) osmotic minipumps were implanted under the skin in the interscapular region. Pumps were filled with carrier alone (subgroups II) plus NPY (subgroups III), or NE (subgroups IV), or both (subgroups V). Delivery rates were calculated to be 0.5 microgram/h NPY; 20 micrograms/h NE, extending over a period of 14 days. Starting from day 2, cumulative food intake and cumulative changes in total body weight were measured every two days. RMR of the animals was measured on days 2, 8, and 14 (indirectly as minimal oxygen consumption). On day 15 animals were sacrificed and specimens of the interscapular BAT were fixed for microscopic examination and measurement of the cross-sectional area of the triglyceride droplets as index for tissue activity. In warm environmental conditions the combined treatment with NE and NPY was the only treatment that caused significant reduction of total body weight by inhibiting food intake and enhancing RMR. The involvement of BAT in this function was observed. In the cold environment the NE and NPY together showed similar but less enhancing effect on body weight; this was mainly due to the significant depression of food intake and slight metabolic response. BAT of this group showed significant response to the combined treatment and slight response to the separate treatments with either NE or NPY. PMID- 9731363 TI - Influence of naproxen on the healing of open excision wound in rats. AB - Excision wounds were made on the paravertebral area on the dorsal side of rat under light ether anaesthesia. The wound was a full thickness wound with removal of the skin upto the fascia. Animals were fed the drug naproxen in a gum acacia suspension with low, median and high doses based on the LD50 value of the drug. The animals were dosed upto the time eschar separated and no raw wound was left. Healing was assessed by planimetric measurement of the wound, periodically, thereby following wound closure by contraction. Granulation tissue formation and wound strength was assessed by using the granuloma model. Epithelialization was also determined. The cellular activity was assessed histologically and the biochemical indices on day of wounding and after the complete healing of the wound. Granulation tissue was used in assessing tensile strength and contraction. Histological sections of the wound on different days were assessed for connective tissue formation by the Mallory-Heidenhan stain. The closure of wounds were represented as percent contraction of original wound size and epithelialization as complete formation of epithelial layer with no rawness of the wound. The scar shape was noted and measured. The results of this study will be discussed. PMID- 9731364 TI - One year histopathological evaluation of fibrous tissue surrounding TCPL implants using adult rats as a model. AB - Fibrous tissue formation is often used as a screening method to determine the biocompatibility of orthopaedic and dental implanted material. In this investigation, porous implants of tricalcium phosphate-lysine delivery devices were implanted intraperitoneally (IP) using adult male rats as a model. The fibrous tissue surrounding the implant was studied histologically to determine the infiltration of inflammatory cells and other healing components. Fibrous capsular tissues were carefully dissected away from the capsule noting the tissue closest to the implanted material. Evaluation of the sections (5um, H&E) collected from various areas of the implants (n = 5 per group) revealed that: (1) The use of TCPL implants resulted in the formation of three distinct cell layers of fibrous tissue. (2) The fibrous tissue thickness was found to be directly proportional to the duration of the implant. (3) The infiltration of macrophages, polymorphonuclear leukocytes, and fibroblasts were present in all phases studied. (4) Ordered collagen was observed only at the end of the 6, 9, and 12 month phases. (5) Remarkable progression of vascularity was present in the second and third layers of the fibrous tissue. In conclusion, these observations confirm our previous finding using different bioceramic material. PMID- 9731365 TI - Preparation and antibacterial evaluation of urinary balloon catheter. AB - Prolonged use of indwelling urinary catheters in bed-ridden patients lead to a variety of Urinary Tract Infections (UTI). The present study deals with the surface treatment of ready-made urinary catheters so as to render them antibacterial in nature. Catheters were treated with varying percentages of iodine in conjunction with natural rubber latex, and their iodide ion release pattern and antibacterial behaviour were evaluated. PMID- 9731366 TI - Biochemical and histological analysis of the flexor tenosynovium in patients with carpal tunnel syndrome. AB - Carpal tunnel release is the most common hand operation performed in this country. In the absence of specific systemic diseases, the etiology and persistence of pain and dysfunction even after surgical decompression is poorly understood. The focus of this investigation was to investigate the biological factors present within the patients serum that may lead to increased sensitivity to pain. Tissue was collected from patients during surgery. The tissue was homogenized and the homogenate analyzed for the presence of IL-1, IL-6, prostaglandin E series (PGE2). The levels were compared with volunteers that had no evidence of carpal tunnel syndrome or pain. The results showed similar levels of IL-1 (range 42-26 ng/ml) in tissue homogenates, and a significant increase in levels of IL-6 and malionaldehyde bis-(diethyl acetal) in CTS patients in comparison to control tissues. This increase may be associated with oxidative changes occurring as a result of ischemia and reperfusion. Tissue homogenates were also evaluated for PGE2. The CTS tissues showed a five fold elevation in PGE2 compared to control tissues. Levels of PGE2 in CTS tissues were statistically different using a two-tailed student T-test. Increased levels of PGE2 can enhance vascular permeability at the site of injury, and can play an important role in activating adenylate cyclase which increases intracellular cyclic adenosine monophosphate (cAMP). This increase in cAMP levels can inhibit functional responses to other inflammatory stimuli. Increases in PGE2 can also cause sensitization of the nerve endings so that a normal stimulus that would not necessarily cause pain will now be experienced as painful. The results of this study demonstrate that arachidonic acid metabolites PGE2 may be responsible for both the pathological changes and clinical symptomatology in carpal tunnel syndrome. PMID- 9731367 TI - Delivery of laboratory data with World Wide Web technology. AB - We have developed an experimental World Wide Web (WWW) based system to deliver laboratory results to clinicians in our Veterinary Medical Teaching Hospital. Laboratory results are generated by the clinical pathology section of our Veterinary Medical Diagnostic Laboratory and stored in a legacy information system. This system does not interface directly to the hospital information system, and it cannot be accessed directly by clinicians. Our "meta" system first parses routine print reports and then instantiates the data into a modern, open architecture relational database using a data model constructed with currently accepted international standards for data representation and communication. The system does not affect either of the existing legacy systems. Location independent delivery of patient data is via a secure WWW based system which maximizes usability and allows "value-added" graphic representations. The data can be viewed with any web browser. Future extensibility and intra- and inter institutional compatibility served as key design criteria. The system is in the process of being evaluated using accepted methods of assessment of information technologies. PMID- 9731368 TI - Hardware and software considerations in 3D ultrasound imaging of a residual limb. AB - An ultrasound scan and data acquisition system has been developed for computer aided prosthetic socket design. The system performs compound scan at each level of the limb by rotating an ultrasound probe around the limb to acquire 36 B-scan images. From these B-scan images, a transverse cross section of the limb is reconstructed using a spatial compounding algorithm. By scanning the limb at many levels, a 3D limb model can be reconstructed. The compound process circumvents the problems associated with the large attenuation of bones and enables reconstruction of a complete image of bones and adjoining soft tissues. In addition, the compound process reduces ultrasound speckle and improves the image quality. These benefits however, can only be achieved through careful hardware and software design. In this paper, methods and algorithms for system calibration, fast and accurate compounding, and removal of the artifacts caused by limb movement during scan are discussed. PMID- 9731369 TI - Data model development as a prelude to implementing a hospital information system. AB - We developed a data model for use in a veterinary teaching hospital using modern object oriented database diagramming. Since we deal with more than one species and often with herds or flocks, the problem became even more complex than the situation in human medicine. We chose to follow, insofar as possible, standards set forth by the HL7 consortium and the American Society for Testing and Materials (ASTM), two organizations deeply involved in setting standards for the health care record. Some modifications became necessary in order to fit the veterinary medical record to these standards but these were straightforward and did not disrupt the overall model. As a first step in constructing a hospital information system we have developed, and present herein, a data model which adequately addresses our needs in both the clinical and financial arenas and which conforms to appropriate standards as completely as possible. PMID- 9731370 TI - Modular data acquisition system updated using LabWindows/CVI Graphical User Interface. AB - The Biomechanics Laboratory of the Neuroscience Department of the Medical College of Wisconsin is currently engaged in research involving trauma biomechanics. For some experiments, 24 channels of analog data must be sampled at 10,000 Hertz. The Modular Data Acquisition System (MDAS) is able to acquire up to 60 channels of analog data at sampling intervals as low as 6 microseconds. This excellent hardware system has only menu-driven utility software that is no longer supported and required updating to a Graphical User Interface (GUI). Using National Instruments LabWindows/CVI software a GUI was developed. The GUI consists of 9 Graphical User Interface panels controlled by a 3000 line C program "MDAS3SAM". "MDAS3SAM" controls a driver program "MDAS_SAM" which communicates with the MDAS unit via a National Instruments GPIB interface. The 9 GUI panels allow the user to configure the MDAS system (selecting channels, sampling interval, triggering levels etc.), start sampling the data, writing the data to hard disk, graphing the data and printing the graphs. The new system allows the user to quickly reconfigure the MDAS unit and obtain accurate results. PMID- 9731371 TI - Application of factor analysis of infrared spectra for quantitative determination of beta-tricalcium phosphate in calcium hydroxylapatite. AB - This work represents a method, which allows to determine phase composition of calcium hydroxylapatite basing on its infrared spectrum. The method uses factor analysis of the spectral data of calibration set of samples to determine minimal number of factors required to reproduce the spectra within experimental error. Multiple linear regression is applied to establish correlation between factor scores of calibration standards and their properties. The regression equations can be used to predict the property value of unknown sample. The regression model was built for determination of beta-tricalcium phosphate content in hydroxylapatite. Statistical estimation of quality of the model was carried out. Application of the factor analysis on spectral data allows to increase accuracy of beta-tricalcium phosphate determination and expand the range of determination towards its less concentration. Reproducibility of results is retained. PMID- 9731372 TI - The materials specialists education and features of biomaterials study and analysis. AB - The work considers some questions of the biomaterials specialists education. The problem of the specialists education in the field of a biomedical material science is not finally solved till now, because of the complexity and variety of necessary knowledge. The main question is what base education is the best for the biomaterial specialists? This work represents the standpoint of scientific school, realizing the educational process based on an electronic engineering material science. PMID- 9731373 TI - Accuracy of fine needle aspiration biopsy of the breast. AB - Fine needle aspiration biopsy (FNAB) has become a widely used and accepted procedure to detect benign and malignant lesions of the breast. In the past, highly invasive procedures were used in the investigation of suspect breast tumors. Surgical excision was the method of choice during this period often requiring hospitalization of the patient. However, the recent introduction of FNAB has allowed a less traumatic approach for such investigations. Due to pressures from Health Management Organizations (HMO's), FNAB is presently the method of choice allowing for decreased patient trauma, expense, and ability of the procedure to be performed on an outpatient basis. Although highly accepted by most clinicians, others suggest that FNAB should perform diagnostically at a level equivalent or higher than that obtainable by frozen tissue sectioning procedures. This study was designed to evaluate the diagnostic ability of FNAB. Four-hundred twenty-seven patients underwent FNAB during 1993-94 at the University of Mississippi Medical Center. Of those patients, two hundred thirty seven also underwent corresponding surgical biopsies. Sensitivity (88%), specificity (96%), a false positive rate (4%), and a false negative rate (12%) were calculated from the data and reported. In conclusion, the diagnostic accuracy of FNAB of the breast determined in this study shows the significance of the procedure. PMID- 9731374 TI - Abnormal cervical lesions in young adults. AB - The objective of this study is to determine the prevalence of cervical lesions in a population of young adults. This study took into account 897,900 pap smears kindly provided by the cytopathology division at the University of Mississippi Medical Center. These cases were subdivided into three different groups based on age. Group one represents the age population of 0-17, group two represents the age population of 18-34, and group three represents the age population of 35 and higher. Seven different parameters were evaluated in this study. These include negative: unsatisfactory, atypia, CIN I, CIN II, CIN III, and invasive carcinoma. Data obtained from this study suggests that cervical lesions among young teenagers were found to be significant and our observations recommend that this population be screened and evaluated on a regular basis. The results of this study conclude that cervical lesions are significant among young adults. This age group would benefit from education and routine screening. In addition early sexual activity, multiple partners, and infectious diseases were noted in cases with cervical cancer. This could be due to the high occurrences of SIL in young adults. Screening of this age group is highly recommended for those young adults at "high risk." PMID- 9731375 TI - Reactive cell change in cervicovaginal smears. AB - Reactive cell change in cervicovaginal smears is a controversial issue. The most common criteria for reactive cell change include an increase in nuclear size, presence of nucleoli, binucleation, cytoplasmic vacuolization, and polychromasia. The purpose of this study is to define, as specifically as possible, the criteria of reactive cell change. Sixty-one cervicovaginal smears in a routine examination obtained during 1988 to 1994 were reviewed for this study. All cases had been diagnosed as reactive. Fifty-three of these were re-diagnosed as reactive and 8 cases were rediagnosed as negative. Inflammatory cells were present in 79% and organisms involvement such as Herpes, Trichomonas, Chlamydia, Gardnerella, and Candida were present in 23% percent. The smears were also evaluated for cellular arrangement, origin of the reactive cells, and presence of nucleoli. The majority of reactive cells were found in aggregates and were of metaplastic origin. Nucleoli were present in 85% of the cases. In all cases the most important criteria of reactive cell change were found to be aggregates of metaplastic cells with central nuclei containing nucleoli and a fine chromatin pattern, followed by the presence of organisms. Additionally, the majority of cases with a cytology diagnosis of reactive cell change had a squamous intraepithelial lesion on biopsy. In conclusion, this study suggests that follow-up Pap smears over a two year period may revert to normal in some of the cases. PMID- 9731376 TI - The effects of image manipulation on automated karyotyping. AB - Karyotyping is one of the standard tools for human genetic investigations. Karyotyping involves the classification and interpretation of chromosomes found in a metaphase spread. As part of the automated karyotyping process, image manipulation is required for appropriately positioning metaphase spread chromosomes in their corresponding karyotype. Image manipulation, specifically image rotation, reorganizes the grey-level information within chromosomes to facilitate analysis. Statistical tests are performed to compare features related to banding pattern and length between unmanipulated chromosomes and corresponding rotated chromosomes. Based on experimental results, reorganizing the grey-level information yields statistically different chromosome features. Depending on the purpose of chromosome image analysis, the interpretation process of karyotyping could be impaired with chromosome feature distortion due to image rotation. PMID- 9731377 TI - Transcutaneous electrovesicogram in normal volunteers, patients with interstitial cystitis, neurogenic bladder, benign prostatic hyperplasia, and after cystectomy. AB - Recordings of the vesical electric activity (electrovesicogram = EVG) were carried out in 22 healthy volunteers (12 male, 10 female: mean age 41.6 years), 9 women with interstitial cystitis (IC: mean age 44.6 years), 24 patients with neurogenic bladder (NB: 16 male, 8 female; mean age 48.3 years), 26 patients with benign prostatic hyperplasia (BPH: mean age 66.8 years), and 9 cystectomy patients (6 men, 3 women: mean age 37.6 years). Four Beckman type silver-silver chloride electrodes were applied to the abdominal skin: one above and lateral to each pubic tubercle, one above symphysis pubis, and a reference electrode to the thigh. Normal EVG manifested as regular slow waves or pacesetter potentials (PPs) which had the same frequency, amplitude and regular rhythm when the test was repeated in the individual subject. Mean frequency was 4.8 cycle/minute (cpm), amplitude 1.7 mV and velocity 4.6 cm/sec. IC manifested with "vesicoarrhythmia" (increased irregular PPs). NB patients exhibited a "dysrhythmic" EVG (irregular PPs) in upper motor neuron lesions and a "silent" EVG in lower motor lesions. In BPH, the compensated bladder showed "tachyvesica" (regular increased PPs) and the decompensated "bradyarrhythmia" (irregular decreased PPs) or "silent" EVG. Cystectomy patients showed a "silent" EVG. In conclusion, transcutaneous EVG may be a useful investigative method in the diagnosis of vesical disorders. It is safe, simple, without complications and cost-effective. PMID- 9731378 TI - Development of a biodynamics work environment (BWE) which integrates a biodynamics data bank, models, and analytical tools. AB - For more than thirty years the Armstrong Laboratory (AL) has studied the response of human volunteers and human surrogates to impact accelerations. Data from these studies form a national resource which should be made available to biodynamics model builders and safety researchers everywhere. Recently AL joined with the National Highway Transportation Safety Administration (NHTSA) to develop a multi media Biodynamics Work Environment (BWE) which will facilitate access to both their respective data bases and provide a means for sharing that access within the scientific community. The BWE concept is sufficiently flexible to allow integration of other databases as well as biodynamics models and tools. The AL data, for example, consists of over 10,000 impact tests which are currently archived on optical media. Each test includes acceleration and motion time histories, subject anthropometry, documentation photos, and high speed video. Among the models to be included are the Articulated Total Body, Head Spine, Dynamic Response Index, Multi-axis Dynamic Response Criteria, and BURNSIM. Complementary analytical tools would involve the use of spreadsheets, statistics, modeling, data analysis, and math packages. This paper will discuss the development, current status, and future plans of the BWE. PMID- 9731379 TI - Accuracy assessment of a technique for contact point determination from planar radiographs. AB - The purpose of this study was to determine the accuracy, precision and repeatability of a simple radiographic method which might be used in in-vivo determination of joint contact points following total joint replacement (TKA). A model of a TKA was constructed from which the anterior/posterior location of the contact point could be mathematically calculated. A radiograph was taken of the model and two observers repeatedly determined the contact point using a parallel rule. The measured locations were compared to the calculated values. The accuracy and precision of the measurement locations in comparison to the calculated values were 1.6 mm and 0.4 mm, respectively. The mean inter-observer error was 0.2 mm. PMID- 9731381 TI - Delay measurement in dual blood volume pulse monitoring using adaptive signal processing techniques. AB - Two adaptive signal processing algorithms are applied to the estimation of the relative delay between Blood Volume Pulse (BVP) signals collected by independent sensors on the subject's index finger. Both the LMS Adaptive Predictor and the Adaptive Delay System are evaluated for the delay estimation in synthetic test signals and on actual pairs of BVP signals. The short-term variations in the relative delay of the BVP signals within a data record is studied and proposed as a key factor determining the better performance of the Adaptive Delay System. Changes in the estimated relative delay between the BVP signals of subjects before and after exercise are shown. It is proposed that these changes are associated with the modifications introduced in the cardiovascular system of the subject by exercise and can, therefore, be used to monitor the level of exercise achieved by a subject at different points in an exercise session. PMID- 9731380 TI - Peripheral nerve (PNS) spinal cord anastomoses bridging spinal cord transection- enhancement of central neurons (CNS) axonal regeneration. AB - These studies involve 76 mature female dogs at 5.5 to 16 kg. The enhancement of axonal regeneration in the spinal cord transection is shown by implanting intercostal nerve with origin cephalic to spinal cord transection. The nerve is implanted in the distal isolated spinal cord near the transection site. The intercostal nerve peripheral portion is anchored in the distal spinal cord using a plasma clot suture. Regeneration of CNS axons and PNS axons in the spinal cord transection demands care, rehabilitation, and maintenance of normal body functions. After a period of two years repair and rehabilitation "reflex" standing and walking developed in 26 mature female dogs. Surgical section of the implanted nerve resulted in loss of standing and reflex walking and return to a paraplegic condition in 12 dogs. Stimulation of the motor cortex and implanted intercostal nerve resulted in movements of the hind limbs of the standing and reflex walking dogs all having cord transection and intercostal nerve anastomoses. PMID- 9731382 TI - Power spectral analysis of autonomic nervous activity in spontaneously hypertensive rats. AB - We studied power spectral analysis of heart rate variability (HRV) in control spontaneously hypertensive rats (SHR) and tourmaline hydroxyl negative ion treated SHR groups. The power spectrum of HRV in SHR groups was composed of two frequency components; low frequency (LF, 0.005 approximately 1.0 Hz) and high frequency (HF, 1.0 approximately 2.0 Hz) components. The low frequency power (LFP) of HRV spectrum, which indicates sympathetic nervous activity, in negative air ion-treated SHR group was significantly (p < 0.01) smaller than in control SHR group. And also we obtained that the high frequency power (HFP) showing parasympathetic nervous activity in negative air ion-treated SHR group was significantly (p < 0.01) higher than in control SHR group. It could be concluded that hydroxyl negative air ions generated by tourmaline ionizer system decreased the elevated blood pressure, and control the sympathetic nervous activity and the parasympathetic nervous activity in SHR having the elevated blood pressure. PMID- 9731383 TI - Artificial vision workbench. AB - Machine vision is an important component of medical systems engineering. Inexpensive miniature solid state cameras are now available. This paper describes how these devices can be used as artificial retinas, to take snapshots and moving pictures in monochrome or color. Used in pairs, they produce a stereoscopic field of vision and enable depth perception. Macular and peripheral vision can be simulated electronically. This paper also presents the author's design of an artificial orbit for this synthetic eye. The orbit supports the eye, protects it, and provides attachment points for the ocular motion control system. Convergence and image fusion can be produced, and saccades simulated, along with the other ocular motions. The use of lenses, filters, irises and focusing mechanisms are also discussed. Typical camera-computer interfaces are described, including the use of "frame grabbers" and analog-to-digital image conversion. Software programs for eye positioning, image manipulation, feature extraction and object recognition are discussed, including the application of artificial neural networks. PMID- 9731384 TI - Review of major injuries and fatalities in USAF ejections, 1981-1995. AB - Our laboratories are examining injuries and deaths resulting from mechanical forces applied to aircrew members in the course of Department of Defense aviation operations. In this paper we report only on bodily injuries sustained during ejection from US Air force, aircraft for the fiscal years 1981-1996, that is, major injuries and fatalities resulting directly from seat acceleration forces, from aerodynamic forces applied to crew members during escape through the effects of windblast and parachute opening shock; from direct contact: and from parachute landing injuries. Such injuries occur typically to the head, neck, cervical spine, thorax, thoracolumbar spine, ribs, pelvis, and the upper and lower extremities. Injuries are usually caused by anomalies in the ejection sequence or by delaying ejection until too close to the ground. Conversely, a planned ejection in a modern ejection seat in controlled, low speed flight imposes forces well below injury thresholds. In the USAF, 10-50 aircrew eject yearly, with a decline since 1991. We conclude that the risk of fatality is 0-11% and of major injury is 2-25%. Both are remarkably low and decreasing in the later years of this study period. The absolute number of head, neck, and spine injuries is 0-10 yearly and similarly decreasing. The results of this study are intended to provide a basis for estimating potential savings in deaths, injuries, and costs expected from the development of improved protective measures. PMID- 9731385 TI - Digital integrated retinal surgical laser system. AB - The year is 2001--ophthalmic retinal surgery is now fully computer assisted. Patients arriving for scheduled treatments of diabetic retinopathy, retinal tears, or macular degeneration have their retina digitally mapped by a technician. From the retinal map, the ophthalmologist plots therapeutic lesion sites with a light pen on the computer screen that will automatically be placed by a computer controlled argon laser. The treatment only requires 100 ms per lesion placement thus reducing office calls to approximately 45 minutes freeing the ophthalmologist for other pressing cases. This paper reports on the development of a clinically significant prototype system that will help bring this scenario to fruition. PMID- 9731386 TI - Co-activation of the hamstrings and quadriceps during the lunge exercise. AB - The anterior lunge exercise is a closed chain kinetic exercise that has been developed to improve the function of the lower limb and to strengthen the hamstrings and quadriceps, simultaneously. In this study, a three-dimensional biomechanical analysis of this exercise was conducted in order to understand the mechanics of this rehabilitation activity. Experimental conditions were recorded using an active optoelectronic kinematic data capture system (OPTOTRAK), two force plates (AMTI) and electromyography (EMG). Data were collected from healthy male subjects while performing several lunges. When the distance between the toe of the rear leg and the heel of the front leg (lunging distance) was maximum, a large net flexion moment was predicted in the front leg in the extented position. This moment was reversed to a large net extension moment in the flexed position. A large increase in the net extension moment in the rear leg was also predicted as the front knee was bent from 5 degrees to 90 degrees of flexion. These data suggest that quadriceps and hamstring muscles co-contraction occur during a maximum lunge in the front leg when it is in the flexed position. PMID- 9731387 TI - Hybrid retinal photocoagulation system using analog tracking. AB - We describe initial in vivo experimental results of a new hybrid digital and analog design for retinal tracking and laser beam control. An overview of the design is given. The results show in vivo tracking rates which exceed the equivalent of 38 degrees per second in the eye, with automated lesion pattern creation. Robotically-assisted laser surgery to treat conditions such as diabetic retinopathy and retinal breaks may soon be realized under clinical conditions with requisite safety using standard video hardware and inexpensive optical components based on this design. PMID- 9731388 TI - The behavior of the hydroxyl groups in polycrystalline porous hydroxyapatite. AB - The activation energy of charge carriers and carrier density in porous polycrystalline hydroxylapatite have been determined by electrophysic measurings in temperature range 15-110'C. As charge carriers in hydroxylapatite is ions, the obtained values were checked by means of IR-spectroscopy using to IR-spectra sections corresponded to oscillations of bond, dissociating to form the conduction ions. It was turned out that mostly the conduction porous polycrystalline patterns of hydroxylapatite has been formed by hydroxylions in the investigated range of the temperature. It is also defined by series of values of activation energy changing from the dissociation energy of water molecule to the split-out-energy of hydroxyl groups of hydroxylapatite molecule. It was observed that values of activation energy and carrier density have been correlated with area of internal surface of material and with amount of hydroxyl groups, the IR-spectroscopy is ascribing to not formula hydroxyl groups of hydroxylapatite. On the basis of these results one is postulated that apatite can exist either in the form of oxy-hydroxylapatite or else in the form of hydroxylapatite, but behavior of share of hydroxylgroups of material is coincide with behavior of hydroxyl groups forming an adsorption lauer of water on the surface of porous polycrystalline hydroxylapatite. PMID- 9731389 TI - Development of a fuzzy logic based system to monitor the electrical responses of nerve fiber. AB - A fuzzy logic based control system to monitor and control the electrical and chemical responses of nerve fibers has been designed and simulated. Fuzzy logic provides powerful reasoning mechanisms for combining information from multiple fuzzy sets.... The rule base has been developed based on the difference between the measured value and the desired value. The potential difference across each nerve fiber and the current through it has been monitored and controlled by a feedback control signal. The controller has been tested and simulated using sinusoidal and square wave forms with different frequency. The desired value was reached very rapidly. The system is capable of monitoring and controlling four nerve fibers simultaneously and allow the researcher to select different modes of operations. PMID- 9731390 TI - Application of artificial neural network for micro-crack and damage evaluation of bone. AB - This paper presents the reasoning and adaptive learning method of artificial neural network (ANN) for micro-crack assessment and damage accumulation due to stiffness loss of dog bone. The importance of using the alternative approach of ANN is that it avoids the complexity of modeling problems, overrides the consideration of simplified assumptions and can be developed directly from experimental data using adaptive learning mechanisms. The proposed artificial neural network model provides a relationship between microdamage accumulation, stiffness loss and number of fatigue cycles (Nf) to failure from an experimental study where stiffness loss and crack area (Cr.Ar., mm2/mm2) are evaluated. This preliminary study using ANN for microdamage evaluation shows that ANN accurately predicts the amount of damage accumulation from stiffness loss. PMID- 9731392 TI - A proposal for a transfer standard for validating pulse oximeter sensors. AB - One method for validating a biosensor is through comparative assessment of the test sensor with a control sensor when both sensors are presented with the same inputs. In the case of adult pulse oximeter (SpO2) sensors, both sensors should ideally be placed on the same digit of the subject. However, at present, one SpO2 sensor interferes with another so that the two sensors are applied to two different digits, which may not be identical in SpO2 level. This article proposes a ring-shaped SpO2 sensor transfer standard that can be used on the same digit as the test sensor, based on the idea of timing, SpO2 is a slowly varying signal, while the two sensors can be gated to measure the same digit's SpO2 value very quickly. Hardware and software requirements for the transfer standard, and a statistical method for analyzing the data will be discussed. A method for the direct comparison of two SpO2 sensors using a "switch over" trial design will also be presented. PMID- 9731391 TI - Effect of aging on human postural control during cognitive tasks. AB - The purpose of this study was to investigate the effect of aging on human postural control during cognitive tasks. Forty-eight subjects between the ages of 20 to 60 years underwent a series of balance tests that placed increasingly more demand on sensorimotor and cognitive mechanisms. Balance tests were based on a dynamic posturography device called EquiTest. The standard clinical protocol consisted of assessing the subject's stability in six different sensory conditions as the inputs from the proprioceptive and visual systems were altered in a systematic manner. The subjects were also tested with the head extended backwards. This represents an additional sensory condition in which the input from the vestibular system is modified. In order to investigate the role of cognitive function on the postural control mechanisms, the balance tests were performed either with or without an attention-demanding task. The test sequence was randomized to account for simple order effects. The relative contribution and interaction of sensory modification, cognitive tasking, and age were assessed. The results showed that head extension significantly increased postural sway in sensory conditions in which the proprioceptive input was altered. The effects of cognitive tasking and age group assignment were only approaching significance. PMID- 9731393 TI - The World Wide Web--a new tool for biomedical engineering education. AB - An ever-increasing variety of materials (text, images, videos, and sound) are available through the World Wide Web (WWW). While textbooks, which are often outdated by the time they are published, are usually limited to black and white text and images, many supplemental materials can be found on the WWW. The WWW also provides many resources for student projects. In BAE 465: Biomedical Engineering Applications, student teams developed WWW-based term projects on biomedical topics, e.g. biomaterials, MRI, and medical ultrasound. After the projects were completed and edited by the instructor, they were placed on-line for world-wide access if permission for this had been granted by the student authors. Projects from three classes have been used to form the basis for an electronic textbook which is available at http:@www.eos.ncsu.edu/bae/research/blanchard /www/465/textbook/. This electronic textbook also includes instructional objectives and sample tests for specific topic areas. Student projects have been linked to the appropriate topic areas within the electronic textbook. Links to relevant sites have been included within the electronic textbook as well as within the individual projects. Students were required to link to images and other materials they wanted to include in their project in order to avoid copyright issues. The drawback to this approach to copyright protection is that addresses can change making links unavailable. In BAE 465 and in BAE 235: Engineering Biology, the WWW has also been used to distribute instructional objectives, the syllabi and class policies, homework problems, and abbreviated lecture notes. This has made maintaining course-related material easier and has reduced the amount of paper used by both the students and the instructor. Goals for the electronic textbook include the addition of instructional simulation programs that can be run from remote sites. In the future, biomedical engineering may be taught in a virtual classroom with participation by an instructor and students from many different parts of the world. PMID- 9731394 TI - A mathematical high time resolution model of the arterial system under extracorporeal circulation. AB - OBJECTIVE: The purpose of the following study was to establish a computer generated model of the hemodynamic effects of pulsatile extracorporal perfusion describing flow and pressure parameters in the body for any given input flow patterns. METHODS: The human arterial tree was delineated according to a 128 branch model encompassing bifurcations and linear physical properties of the arterial walls. The distribution of flow and pressure waves was calculated based on a refined 3-element windkessel model. Autoregulatory mechanisms of brain and kidneys were implemented. RESULTS: By providing a simulated, "pump-generated" flow curve as the input signal to the system, the model was able to create and display flow and pressure curves at a high time resolution in each part of the systemic circulation including reflection phenomena throughout any observation period chosen. The hemodynamic effects of different pump-flow patterns, age, variations in hematocrit, hypothermia and occlusion of arterial branches, like the renal artery, could be simulated. CONCLUSION: In an attempt to get closer to a mathematically based regulation of heart-lung machines, this model of computer generated extracorporeal circulation provides an initial step. Ongoing research is required for implementation of metabolic conditions and continuous approximation of the model of the real physiologic or pathologic situation. PMID- 9731395 TI - Putting the shoulder to the wheel: a new biomechanical model for the shoulder girdle. AB - The least successfully modeled joint complex has been the shoulder. In multi segmented mathematical shoulder models rigid beams (the bones) act as a series of columns or levers to transmit forces or loads to the axial skeleton. Forces passing through the almost frictionless joints must, somehow, always be directed perfectly perpendicular to the joints as only loads directed at right angles to the surfaces could transfer across frictionless joints. Loads transmitted to the axial skeleton would have to pass through the moving ribs or the weak jointed clavicle and then through the ribs. A new model of the shoulder girdle, based on the tension icosahedron described by Buckminster Fuller, is proposed that permits the compression loads passing through the arm and shoulder to be transferred to the axial skeleton through its soft tissues. In this model the scapula 'floats' in the tension network of shoulder girdle muscles just as the hub of the wire wheel is suspended in its tension network of spokes. With this construct inefficient beams and levers are eliminated. A more energy efficient, load distributing, integrated, hierarchical system is created. PMID- 9731396 TI - Production of spiral fractures in human cadaver long bones by use of a simple torsion machine. AB - A simple machine was developed so that long bones could be torqued to failure in order to estimate applied moment and study the resulting spiral fracture pattern. Detailed study of the fragments enabled the postulation of an orderly manner of fracture propagation. Twenty-seven embalmed human cadaver femurs were studied. The machine held one end of the bone in a fixed vise and the other in a rotating vise. Spring scales were used to apply force to a bar connected to the rotating vise which allowed for a crude estimation of torsional moment. Male femurs (72.8 yrs old sigma = 11.4) fractured at a mean torque of 106.7 N-m (sigma = 23.8), while female femurs (78.0 yrs old sigma = 6.7) failed at a mean torque of 96.7 N m (sigma = 39.4). There were no statistically significant differences between the ages (p = 0.20) or the torsional moments at fracture (p = 0.41) for the male and female femurs. Given the similarities in this study population, an eye was turned towards anthropomorphic measurements in order to determine factors that might be indicative of bone strength. Several measurements were made on the fragments especially in the midshaft region. These included six cortex thicknesses, bone depth, width and circumference. Simple statistical analyses were performed. PMID- 9731397 TI - Fractures of experimentally traumatized embalmed versus unembalmed cadaver legs. AB - Intact legs from six geriatric cadavers were fractured in a self-controlled study aimed at documenting the effects of embalming on both the soft and hard tissues of cadaver specimens subjected to biomechanical impact research. Upon bequeathal, one leg was removed and frozen while the other remained with the cadaver for embalming. The embalmed legs were amputated later and pre-test radiographs were made. For testing, a rod was inserted in the upright leg such that simulated upper body mass could be applied. A 50 kg cart propelled by a pneumatic accelerator to 7.5 m/s struck the anterior leg midway between the knee and ankle. The cart was headed by an instrumented steel pipe (4.75 cm dia.) coupled to a transducer which relayed impact force data to a Hewlett Packard 3562 A signal analyzer. Testing was captured on standard VHS video (30 frames/s) and 16 mm Color High Speed Film (1,000 frames/s). Post-test analyses included radiographs and thorough dissection. Peak forces were comparable for matched pairs. The unembalmed legs showed greater soft tissue damage (muscle and skin) but generally less bone fragmentation than their embalmed counterparts. Neurovascular components were virtually unharmed in most legs. PMID- 9731398 TI - Medical and financial aspects of same-day bilateral total knee arthroplasties. AB - In this retrospective study, we compared forty-five patients who had undergone same-day bilateral total knee arthroplasties with 144 patients who had undergone unilateral total knee arthroplasty procedures. All procedures were performed by a single surgeon using a single prosthetic design between June 1994 to May 1996. The issues studied were the length of hospital stay, hospital charges, units of blood transfused, units of homologous blood transfused, and the rate and type of complications. This study also examined the usage and financial charges of extended care facilities and/or home health care following discharge from the hospital. Results show that, for patients suffering from symptomatic bilateral knee disease, the same-day bilateral total knee procedure is significantly less costly than staged bilateral procedures. The rate and type of complications seen in the same-day bilateral cases were not significantly different from complications seen in unilateral cases. PMID- 9731399 TI - Effects of limb perturbation on surface recorded nerve action potentials. AB - The field action potentials are a convolution of source strength and the propagating-point source responses (PPSRs). Surface-recorded peripheral nerve potentials are strongly influenced by the volume-conductor properties of the limb. In this paper the PPSRs are solved based on finite-element formulation, which allows solutions to forward volume-conductor problems involving curved nerves in arbitrarily shaped, inhomogeneous, anisotropic limbs. Effects of limb perturbation on surface-recorded nerve action potentials are simulated based on circular and elliptic cylinders with a uniform cross-section consisting of multiple inhomogeneous and anisotropic regions (fat, muscles, and bone layers). The simulations show that a simple homogeneous model is adequate only for a very superficial nerve that lies completely in the subcutaneous fat layer. Deeper nerves that lie on or within the muscle require a multiple-layer model that takes both the muscle and the fat into account. The cross section only needs to be modeled accurately between the recording site and the nerve location. A circular limb model is adequate for clinical recordings. However, an accurate knowledge of the nerve path is essential for accurate modeling of the action potentials, in both needle and surface recordings. PMID- 9731400 TI - A neural network approach to markerless measurement of human motion. AB - Most automatic motion analysis system operates by tracking markers across a field of view. The markers are usually attached to the skin at the joints. For some applications such as measuring the motion of elderly and the disabled persons, this approach can be uncomfortable which ultimately can affect the motion itself. The protocol of some markers can also be a problem for example active markers using infra-red LEDs must continuously be in the field of view of the detectors at all times. This will impose restriction in the motion. Therefore a marker free method is an attractive proposition. Such an approach would allow the subject to move freely thus exhibiting a more natural movement. We have developed a PC based system which uses frames from the video or photographic capture of a motion in the frontal and sagital plane. Key frames are selected and co-ordinate positions of the joints are identified. Joint angles are calculated from these positions based on a multiple link model of the human figure. The angles are then used as training data for a fully connected neural network. The neural network generates a model of the captured motion which is used to reproduce closely the actual motion. PMID- 9731401 TI - Development and clinical evaluation of Chitra visible light cure composite. AB - Two paste, self cured BIS-GMA based resin was developed and physicochemical and biological characterization carried out. Its suitability and safety for clinical use were evaluated. Clinical trials for direct bonding of Orthodontic brackets conducted at the College of Dental Surgery, Manipal showed a success rate of 83% against 85% with "Right On" adhesive used as control. Further research to improve shelf life of the material led to the development of light cure-BIS-GMA based resin. It was evaluated for physicochemical characteristics which included setting time, compressive strength, diametral tensile strength, microhardness, water sorption and exothermic reaction. Biological characterization studies on material toxicity and biocompatibility on animal models established the safety and suitability of this composite for clinical use in dentistry. Clinical evaluation for direct bonding of Orthodontic brackets in comparison to Ultralight (T.P Orthodontics, USA) showed its handling characteristics to be superior to control. Flash removal was easier around bracket bases bonded with Chitra light cure. However, bond strength of light cure composite was 66% as compared to 79% with Ultralight. Subsequent improvement of the material increased the bond strength significantly. Failure rate with the new material was 11% against 13% with control in the ongoing clinical trials. PMID- 9731402 TI - Knee kinematics in-vivo of kneeling in deep flexion examined by bi-planar radiographs. AB - Squatting and kneeling are important daily activities for Middle and Far East cultures that require positioning the knee in deep flexion. In these activities, the limb becomes fully flexed with a knee flexion angle reaching between 150 and 160 degrees and the heel reaching the posterior surface of the upper thigh. Existing knee prostheses do not allow a full return to normal activities for this large population since they are limited to achieving knee flexion of about 120 degrees. Also, there is very limited information on knee kinematics and/or forces in the range beyond 120 degrees. The purpose of this study is to describe the kinematics of normal knees in-vivo, assessed in deep flexion, using bi-planar radiographs. A-P and lateral views were obtained from 5 healthy subjects during three sequential positions of kneeling. In the 1st position, the subject knelt with the knees fully flexed (deep flexion between 150 degrees and 160 degrees) and torso upright. In the 2nd position, the subject bowed forward to an intermediate position (about 120 degrees of knee flexion). In the 3rd position, the subject bowed further until his/her head touched the floor, supporting the upper torso with hands and attaining a knee flexion of about 90 degrees. The results show that past 135 degrees of knee flexion, the patella was found to clear the femoral groove and was in contact only with the condyles. The results also show that the classical femoral "roll back" does not appear to occur in deep flexion. It seems that the lateral femoral condyle rolls over the postero medial aspect of the lateral tibial plateau while contact of the medial femoral condyle occurs more anteriorly, but still in the posterior aspect of the medial tibial plateau. This asymmetric rolling motion implies an element of internal tibial rotation. Furthermore, the tibia was found to articulate with the femur at the most proximal points of the condyles in deep flexion. These data on the kinematics and contact characteristics of the tibio-femoral joint must be considered in any approach to design for a Deep Flexion Knee Implant. PMID- 9731403 TI - Validation of the advanced dynamic anthropomorphic manikin (ADAM) database: horizontal sled test. AB - As the U.S. Air Force (USAF) continues to introduce advanced technology to make its planes more dynamic, it is becoming increasingly more difficult to adequately test the systems to ensure pilot safety. A cost effective solution to this problem is the use of computer modeling to augment testing. The accuracy of such computer modeling depends on the validity of the analytical formulation, and the correctness of the database characterizing the systems being modeled. One such database is for the large Advanced Dynamic Anthropomorphic Manikin (ADAM); a human surrogate developed by the USAF for high speed ejection testing. The database is used in the Articulated Total Body (ATB) computer model utilized by the Armstrong Laboratories to predict human body dynamics during aircraft crashes and emergency escapes. The large ADAM database, and the parameters from a horizontal sled test were used in an ATB sled simulation. The results of the ATB simulation are compared with actual sled test data. These results include head, chest, and pelvis accelerations; neck and lumbar loads; and elbow, knee, hip and shoulder angular motion. The comparisons are the basis for validating the ADAM database for future predictive simulations. PMID- 9731405 TI - Quadriceps femoris function during knee extension following total knee arthroplasty. AB - Electromyographic signals (EMG) from surface electrodes over the vastus medialis, rectus femoris and vastus lateralis were monitored during isometric knee extension for 10 TKA patients and 6 control subjects. No significant side-to-side differences in normalized EMG signals from any of the monitored muscles were found when the left and right legs of the control group were compared or when the operative and the non-operative legs of the patient group were compared. However, both the operative and the non-operative legs in the patient group differed significantly (p < 0.01) in normalized EMG from the control group. This study has shown that a muscle imbalance, possibly leading to patellar tracking problems, does not routinely exist following TKA through a medial parapatellar incision. PMID- 9731404 TI - Validation of electrolytic-liquid tilt sensors for human motion measurement. AB - The development of inexpensive, low-weight, no-invasive sensors that can be used to accurately measure human motion is important for the evaluation of biomechanical risk during a variety of industrial work activities. Electrolytic liquid tilt sensors that measure the angular position of an object relative to the gravitational force vector may provide a valuable means for assessing the postural demand of work tasks. For example, the biomechanical cost to the low back could be assessed during manual material handling activities, or the demand to the shoulders could be evaluated during overhead construction work. Tilt sensors were tested during static and dynamic activities, using isovelocity and isoacceleration dynamometers to move the sensor. The output voltage of the sensors was found to be linearly proportional to the angular deviation of the dynamometer within an 150 degree range (r > 0.99). During constant angular velocity and constant angular acceleration movements, the correlation between the output of the dynamometer and sensor was high and linearly dependent on angular position. Hence, the sensors are capable of accurate motion measurement during static and controlled dynamic movements. Because of the inertia of the liquid within the sensor, its output during sudden acceleration/deceleration may cause some artifact which requires more extensive investigation. PMID- 9731406 TI - Selected comparisons between experienced and non-experienced individuals during manual wheelchair propulsion. AB - The present study was undertaken to identify potential errors that might arise from combining able-bodied individuals with wheelchair dependent individuals to form larger sample sizes when studying manual wheelchair propulsion. Five able bodied and five wheelchair dependent individuals, ranging in age from 24 to 36, propelled a wheelchair on a dynamometer at three different velocities. Of the nine variables (at three different velocities) investigated, significant differences (p < 0.05) were obtained for three physiological characteristics: propulsion efficiency, peak oxygen consumption, and energy input all at high velocity, and for four technique characteristics: at low velocity (propulsion time), and at high velocity (propulsion time, push angle, work per stroke). The results suggest that able-bodied individuals should not be used to increase the size of subject pools when doing studies which include these variables. PMID- 9731407 TI - Heart rate variability in the horse by ambulatory monitoring. AB - Using a microprocessor controlled Ambulatory Monitoring System (AMS) developed by one of us (LvD), we have been studying the changes in and control of heart rate in the resting horse. The system provides us with InterBeat Intervals (IBI in milliseconds), motion sensing, and a time domain measure (mean successive differences: MSD) of heart rate variability for periods up to 72 hours. Thoracic impedance is also available but parameters for the equine chest are not currently available. The system is completely noninvasive, small, and carried on a surcingle worn by the subject. The equine subject is confined to a stall in our teaching hospital but not otherwise restrained. Recording is virtually unobtrusive. Ten horses (judged to be clinically normal) were used in this preliminary study. After collection, the data were "offloaded" to a laptop computer for additional analysis. The electrocardiogram could be recorded on each of the ten animals. Complete data, suitable for spectral analysis, were obtained from four of the animals. Spectral estimates were calculated on periods of varying lengths (3-5 minutes) with more stable spectral estimates associated with longer recording periods. Results indicated the preponderance of parasympathetic control in equid heart rate. These results provide support for the utility of this method for the study of heart rate variability in the freely behaving horse. PMID- 9731408 TI - Optimizing muscle-to-cardiac timing for aortomyoplasty. AB - A new surgical approach to support failing hearts is known as aortomyoplasty-a technique in which the latissimus dorsi muscle is wrapped around the aorta and stimulated during cardiac diastole to provide chronic diastolic counterpulsation. We hypothesized that the timing of muscle contraction within the cardiac cycle effects the amount of diastolic augmentation during counterpulsation. In dogs (n = 9, 20-25 kg), the effect of muscle-to-cardiac timing on hemodynamic outcome of aortomyoplasty was measured. Muscle stimulation was initiated at the dicrotic notch and stimulus durations were systematically increased. Mean diastolic aortic pressure was maximized when stimulation terminated at the ensuing R-wave. Peak left-ventricular pressure was minimized when muscle stimulation terminated before the ensuing R-wave. The endocardial-viability ratio (a ratio of aortic diastolic pressure augmentation to left-ventricular pressure reduction) was maximized when stimulus terminated at the ensuing R-wave. Muscle-to-cardiac timing influences the effectiveness of counterpulsation during aortomyoplasty. PMID- 9731409 TI - Estimation of the correlation dimension of heart rate using surrogate data techniques. AB - Researchers have recently applied nonlinear dynamical systems theory to physiological time series data. However, the direct implementation of the deterministic equations used to estimate the correlation dimension is problematic for physiological data. Importantly, the distinction between a nonlinear process and noise is particularly difficult for experimental data. One technique that has been developed to help with this distinction is the use of surrogate data. These data preserve many of the properties of the original time series but in fact are a randomization of the original data. Whereas in a sufficiently high embedding space the estimates of the correlation dimension of a nonlinear process will plateau, such estimates for a noise process will continue to rise with increases in the embedding dimension. In the present experiment two data sets were examined. Heart period time series were collected from a female subject during spontaneous breathing and breathing paced at 15 breaths per minute. Both data sets consisted of approximately 3000 interbeat intervals and embedding dimensions of m = 2 through m = 6 were examined. Results indicate that the correlation dimension estimate for the spontaneous breathing data reached a plateau around 4, whereas the surrogate data for this series showed no such plateau. For the paced breathing data, both the original and the surrogate data failed to show evidence of a plateau in the estimates of the correlation dimension. These results suggest that caution must be used in interpreting finite values of correlation dimension as evidence of a chaotic attractor in experimental data if no test for determinism is performed. PMID- 9731410 TI - Wavelet analysis of SAECG to identify patients with conduction defects at risk for sudden cardiac death. AB - The aim of this study was to determine how Wavelet transform analysis of signal averaged ECGs can identify patients with conduction defects who are at high risk for development of ventricular tachycardia. In this study, 34 SA-ECGs and programmed electrical stimulation (PES) reports were obtained from the OSU Department of Cardiology Database (1988-1996) and divided into two groups: 17 patients that had inducible monomorphic VT by PES (VT+) and 17 that showed no arrhythmias (VT-). We used Morlet's wavelet to analyze the X, Y, Z, and RMS vector magnitudes in each group. The mean duration from the peak of the RMS vector magnitude to the QRS offset was statistically different with a T value (2 tailed distribution, unequal variance) of 0.033. We noted statistically significant (p < 0.0001) differences in Wavelet energies for 44 msec after the peak of the RMS vector magnitude largest in the Z lead, the first 22 msec, and frequency bins less than 131 Hz. Although no clinical marker could be determined using Wavelet analysis to distinguish the the VT+ from the VT- group, the results from this study show that their SA-ECGs are indeed different even though the optimal analysis has not yet been devised. PMID- 9731411 TI - Improving portable inhalation therapy through real-time assessment of patient pulmonary state. AB - Metered-dose inhalation therapy is the method of choice for a number of respiratory conditions including asthma and bronchitis. Correct use of the metered-dose inhaler (MDI) has been the subject of much research. Devices have been developed to assist with training in their proper use and to enhance the effectiveness of MDI therapy. However, all lack patient pulmonary state assessment. We have approached the development of an improved portable MDI device by focusing on the total system elements that contribute to a smart inhalation therapy device. Components of such a system should include assessment of the patient's therapeutic state, tracking of drug usage, and also include physician modified alarm thresholds. Therapeutic state is best assessed with end-expiratory CO2 measurements augmented by inspiratory and expiratory flow efforts. Drug usage monitoring needs to include the time history of drug administration. Physician modified alarms should allow configuration of the smart inhaler to account for patient condition so that appropriate alarm limits can be established; for example, raising the CO2 alarm threshold for patients with chronic lung disease. PMID- 9731412 TI - Homohemodialysis: a technique of dialysis for the uremic animal. AB - As an alternative to kidney transplantation in conditions of renal failure, an in situ kidney of a healthy individual may be used. The present study utilizes the kidney of a healthy rabbit as a hemoperfusion unit for another, uremic, animal. The study comprised 17 experimental models, each of which consisted of 2 adult New Zealand rabbits. One animal of each model was rendered uremic by means of bilateral nephrectomy. The blood chemistry (urea, creatinine, sodium, potassium, pH and base deficit) was examined pre- and 4-hourly post-operatively. When after 48 hours post-nephrectomy, the blood chemistry had reached a level sufficient to endanger the animal's life, homohemodialysis was performed. The anticoagulated blood was circulated from the uremic animal to the normal one and then back to the uremic animal through the femoral vessels using a tube system. The blood chemistry was determined every 10 minutes and pH and base deficit every 30 minutes. All animals died or were sacrificed within 21 hours after shunting was started, and autopsy was done. Serum sodium was the first to normalize within the first 10 minutes post-shunting, followed by serum potassium and pH in 30 minutes. Blood urea and creatinine reached normal levels in 40 minutes and base deficit in 60 minutes post-shunting. The pathologic examination of specimens from the vital organs of both the normal and uremic animals showed different degrees of cellular damage probably due to hypotension or acute effects of the unbalanced animal homeostasis. The cellular damage was much less in the normal than in the uremic animals. In conclusion, homohemodialysis proved to be effective in normalizing the concentrations of the different substances retained in the blood of uremic animals within only 60 minutes of dialysis. PMID- 9731413 TI - Response of human fibroblasts to tantalum and titanium in cell culture. AB - The loosening of total joint arthroplasties (TKA) with associated osteolysis has been a persistent problem in orthopaedics. Wear debris from prosthetic devices including Titanium (Ti) is involved in this process. Mechanisms for this osteolytic process are unclear. The purpose of this study was to compare the biological response of Ti and Tantalum (Ta) on retrieved human fibroblasts. Fibroblasts were retrieved from human volunteers and cultured using standard techniques. Twenty-five (25) ml culture flasks were seeded with cells and when reaching confluency four concentrations of Ti and Ta were added. Their mean size was less than 3 microns for both metals and gram weights were 0.0048. 0.0096, 0.048, and 0.096 gms. After ten (10) days the cells were fixed, stained and photographed. For both Ti and Ta, the lowest concentration had little effect on the cells, while at the two higher concentrations, nearly all of the cell were killed. Since both of the metals tested are considered to be inert with respect to toxicity, these results would suggest that the observed cell death, seen equally for both metals, was due to the size and concentration of the particles and not to the metals tested. Mechanisms are currently being investigated which include mechanical as well as chemical factors. PMID- 9731414 TI - A comparison of interleukin-1 beta in human synovial fluid of osteoarthritic and revision total knee arthroplasty. AB - Interleukin-1 beta levels from synovial aspirates were examined to determine if they can be used as indicators of increased synovial activity and an inflammatory reaction within total knee arthroplasty. Synovial aspirates were obtained from twelve osteoarthritic knees scheduled for total knee arthroplasty and twenty-one knees scheduled for total knee revision. Eleven of the revision cases involved titanium alloy prostheses and ten involved cobalt-chrome prostheses. Using a high sensitivity ELISA test kit, the interleukin-1 beta concentrations were compared. A significant difference in the interleukin-1 beta concentration was found between all three knee groups. The knees scheduled for revision surgery showed higher concentrations of interleukin-1 beta than osteoarthritic knees. While the knees implanted with a titanium prosthesis showed the greatest concentration of interleukin-1 beta, the osteoarthritic knees showed the lowest interleukin-1 beta concentration. Furthermore, significantly greater synovial aspirate volumes were obtained from the revision cases than from the osteoarthritic cases. This increased synovial activity is most likely attributed to the high concentration of the particulate wear debris produced from the prosthesis. PMID- 9731415 TI - Frequency dependent radial compliance of latex tubing. AB - Custom latex tubing is often used in medical device evaluation. Examples include thin-walled devices used to reduce leakage of porous vascular grafts, and thicker walled prototypes used as mechanically equivalent synthetic arteries. Medical devices such as stents and balloons are introduced into these for mechanically comparable in vitro testing. The three-dimensional mechanical properties of these tubes are critically important, particularly in accelerated testing, since they are primarily designed to replicate the mechanical rather than biological properties of in vivo arteries. This paper explores the instrumentation and protocols necessary to evaluate the frequency dependent radial compliance of precision built latex tubing. Five cm long samples of custom dipped latex tubing 6 mm in diameter with wall thickness from 0.015" to 0.033" were kept dry or soaked in 37 degrees C phosphate buffered saline for 48 or 96 hours before being mounted on a dynamic internal compliance tester. Each tube was tested initially at 70 bpm to establish the internal radial compliance at the physiologically relevant rate. The frequency of the test was then increased incrementally and the radial compliance re-checked. In the most extreme case, tubes were tested up to 2700 bpm. In each case, the volume, pressure, and length of the tube was monitored continuously. PMID- 9731416 TI - Biochemical and immunochemical evaluation of tissues and synovial fluid from patients undergoing total joint arthroplasty. AB - Cytokines are inflammatory mediators responsible for numerous clinical conditions, and are thought to lead to the resorption of bone. Understanding the nature of the cells producing these factors which control the resorption of bone will ultimately lead to a better understanding of why implants fail or integrate. In this study, synovial tissues and synovial fluids were processed for biochemical as well as histochemical and immunohistochemical determination cytokines responsible for bone resorption. The results from this study showed by both quantitative enzyme linked immunoassay (ELISA) and qualitatively by immunohistology a marked increase (twofold) in interleukin-1 (IL-1), and tumor necrosis factor-beta (TNF beta) in synovial tissues in comparison to control tissues of cartilage, ligament and meniscus. Evaluation of tissues both immunochemically and by Hematoxylin and Eosin demonstrated the presence of fibroblast and cells such as macrophages, and multinucleated giant cells in the synovium that are capable of producing bone resorption. Synovial fluid from primary and revision patients were evaluated for TNF beta and IL-1 were not statistically different. Overall, the results indicate that the inflammatory cells of the synovium are secreting factors which may act to mediate aseptic loosening of implants. PMID- 9731417 TI - The effect of biological media on the hydrolysis of mustard simulants. AB - A useful decontaminant for mustard would be one which is readily available in large quantities, inexpensive and potentially biodegradable. Currently one that is being investigated consists of a mixture containing bovine hemoglobin, gelatin and poi. Two of them, gelatin and poi, are common foodstuffs in the diet on the mainland of the United States and in the Hawaiian culture, respectively. Various combinations of these substances have unusual flow properties since the physical states change from a liquid to a gel form. The hydrolytic kinetics of the mustard simulants, 2-chloroethyl ethyl, 2-chloroethyl methyl and 2-bromoethyl phenyl sulfide are reported. The kinetic mechanisms and rate constants are dependent upon the mixtures' concentration and viscosity. PMID- 9731418 TI - The differential expression of NGFS-like substance from fresh pilose antler of Cervus nippon Temminck. AB - Extracts of cartilage from antlers of non-mature deer, e.g. Cervus nippon Temminck, have been used as Chinese traditional medicines and restoratives. Since bioactivities attributed to the antler extracts resembled growth factor activities, the objective of our study was to compare the biological activities of different deer antler extracts with those of known growth factors. Extracts of the deer antlers were found to stimulate the growth of nerve fibers and to induce morphologic changes during differentiation and effect DNA synthesis in PC-12 cells, thus sharing with a known growth factor, NGF, several bioactivities. PMID- 9731419 TI - Lipoprotein activated macrophages in co-culture with mesangial and endothelial cells: nitric oxide and tissue injury. AB - Experiments were conducted to test the hypothesis that VLDL induced excess NO in macrophages may be the mechanism of tissue injury that explains the risk associated with VLDL towards the development of cardiovascular and renal diseases. VLDL activated macrophages were co-cultured with endothelial and mesangial cells. The results showed 59-63 fold higher NO production in VLDL activated macrophages compared to the unactivated macrophages. Nitric oxide production was further potentiated by an additional four fold (241 fold) in VLDL activated macrophage/mesangial cell co-cultures. In co-cultures of VLDL activated macrophages/endothelial cells, NO production was 47 fold higher than the corresponding controls. Cell injury as measured by lactate dehydrogenase activity in cell culture supernatant was elevated in co-cultures of VLDL activated macrophage and mesangial or endothelial cells. Endothelial cells were more susceptible to nitric oxide mediated injury than mesangial cells. On the other hand, low density lipoprotein (LDL) activated macrophages did not generate significant NO or exert toxicity. These results provide support for the mechanism by which VLDL can cause tissue injury and highlights increased nitric oxide production due to the interaction between VLDL activated macrophages and mesangial cells. PMID- 9731421 TI - The use of the tunica vaginalis sac as a dialyser for the uremic animal model. AB - The tunica vaginalis (TV), being part of the peritoneum, can be used for dialysis. The present study explores the possibility of using the TV sac as a dialyser in 21 male adult New Zealand rabbits which had been rendered uremic by ligation of both ureters. 4 animals acted as controls and another 4 as sham controls. Blood chemistry including blood urea, creatinine, sodium and potassium was assessed pre- and 12 hourly post-ureteric ligation. Intratunical dialysis was started 48 hours after ureteric ligation when a high uremic state had been achieved. Blood chemistry was determined 4 hourly post-dialysis. Dialysate was replaced by a fresh one when the blood chemistry was no further improving. After blood chemistry normalization, the animals were sacrificed and autopsied. Blood chemistry improved after TV dialysis in all rabbits and normalized in 13. Eight animals, in spite of showing improvement in blood chemistry, died before normalization. Blood chemistry normalization necessitated dialysate replacement by a fresh one more than once (3 times in 11 rabbits and 4 times in 2). Autopsy showed degenerative changes in kidneys, liver, spleen, lungs and small intestine which were less marked in the animals whose blood chemistry had normalized. In conclusion, intratunical dialysis succeeded in normalizing the blood chemistry of uremic animals. PMID- 9731420 TI - Effect of dehydroepiandrosterone on endothelial cell proliferation. AB - The steroid dehydroepiandrosterone (DHEA) is know to have anti-atherosclerotic properties. However, the mechanism by which DHEA exerts its effect is not clear. We hypothesized that DHEA protects against the atherogenic effects of LDL. Therefore attempts were made to test this hypothesis on endothelial cells (EC) in culture. Experiments were performed to determine the effect of DHEA and LDL on EC proliferation. Incubation of DHEA with EC for 24 hours had minimal effect on cell proliferation. However, at 48 hours of incubation, 25 and 50 nanograms DHEA inhibited EC proliferation by 50%. LDL (20 ug) inhibited EC proliferation by 27% and 63% at 24 and 48 hours respectively compared to the corresponding controls. EC proliferation was stimulated by 40% in the presence of 5 ng of DHEA and LDL in 24 hour incubations. However, at 48 hour incubations DHEA negated the inhibitory effect of LDL on EC proliferation. Determination of malondialdehyde (MDA), a measure of oxidative damage shows that in incubations of EC containing 5, 25, and 50 ng of DHEA for 24 hour did not affect the MDA levels. However, at 48 hour incubations, MDA levels were 44% and 50% higher respectively compared to the control. MDA levels increased in EC cultures containing LDL by 259% and 208% at 24 and 48 hours respectively compared to the corresponding controls. EC cultures containing LDL and 5 ng DHEA reduced MDA levels by 62% and 50% at 24 and 48 hours respectively compared to the corresponding controls. These results show that DHEA protects EC against LDL induced cytotoxic effect and suggests a mechanism for anti-atherosclerotic effect of DHEA. PMID- 9731422 TI - Animal experimentation with tooth derived calcium hydroxyapatite based composites as bone-graft substitute biomaterials. AB - Calcium Hydroxyapatite (HA) has now been clinically accepted as one of the most important alloplastic bonegraft substitute in the management of oral surgical procedures including periodontal bony defects. HA can be synthetically prepared or derived from natural sources. Among the natural sources bovine bone is common but human source is also used. Of late Calcium Hydroxyapatite has been derived from extracted human tooth source. The present study involves the experiments using different varieties of HA in conjunction with chitosan, a binder, to know it's unique biological behaviour of bone bonding. The experiment was designed using 12(Twelve) New Zealand white rabbits on transcortical drilled hole tibial model and the observation was made under optical and scanning electron microscope. Result showed the osteoconductive and bioactive nature of tooth derived HA and biocompatibility of chitosan. The clinical problem of handling the particulate form of HA can be overcome by using chitosan which when mix with HA can stabilize the particle in surgical sites. This has tremendous clinical applications in dental regenerative surgery. PMID- 9731423 TI - Flow analysis of red blood cell through microvascular bifurcations. AB - The motion of a rigid particle suspended in two-dimensional flow through bifurcations of diverging microvessels (DMs), and converging microvessels (CMs) is dependent on multiple complex factors that include the geometry of the network, the angle of the bifurcation, the pressure gradient across the microvessel, the geometry of the cells and their radial location in the vessels. To determine how these parameters affect cell trajectories and flux into downstream branches of CMs and DMs, the motion of cells flowing into a DMs and CMs bifurcation, with a 45 degrees branch angle has been modelled, for every location by means of the finite-difference analysis (FDA). The modeling data was compared with direct experimental data from converging and diverging microvessels obtained from mesenteric microcirculation of the rat. Detailed statistical analysis showed significant correlation between the modeling data and experimental data. This model provides estimates of RBC flow along the trajectory path through bifurcations of CMs and DMs; sites which may be crucial for flow stoppage, depending on the vessel diameter and cell deformability. PMID- 9731424 TI - Equivalent tree representation of electrocardiogram using genetic algorithm. AB - Electrocardiogram (ECG) gives the electrical activity of the heart. The number of data points required to represent the ECG signal is reduced by using a complete tree representation. This reduced data structure (ECG Tree) is obtained by fitting the ECG signal in a grid structure consisting of both horizontal and vertical lines. The leaf nodes are the points where the vertical grid lines intersect with the ECG signal. These leaf nodes now form the features of the ECG signal. Some of these leaf nodes may be redundant and hence the reduction in the number of leaf nodes and thus optimization of the tree (equivalent tree) is done using a novel technique based on the Genetic Algorithm (GA). In this work, the equivalent tree is formed using GA consisting of four stages. First, from the group of generated leaf nodes various combinations of strings are constructed to form the population. Second, the fitness function is taken as the measure of the vertical distances between two neighbouring leaf nodes in order to evaluate the population with respect to their fitness values. Third, the selection procedure is used to give offsprings based on an assigned threshold value. Finally, crossover and mutation operations are performed repeatedly till an optimized population is obtained. The optimal nodes represent the equivalent tree. The Backpropagation Neural Network as a classifier is used to test the efficacy of the GA in this optimization problem. PMID- 9731425 TI - Visualization of a calcium hydroxylapatite molecular structure. AB - The creation of the three-dimensional complex spatial structure, and peculiarity of changes occurring in a material, because of external influence. The algorithm of the volumetric (space-filling) and ball-pivot model of molecules images construction applied in the system, allows to display ball-pivot and volumetric models of molecules movement in real time scale. Pseudo-three-dimensional image of a molecule, allowing a good look at it from the different points of view, is formed and the opportunity of the stereoimage creation increases presentation effect of its volume many times. The work contains the standard crystal lattice parameters account method of a calcium hydroxylapatite based on the rontgenogram of the material samples. The summary table of Cartesian coordinates of atoms of a calcium hydroxylapatite is applied. PMID- 9731426 TI - Study on plasma-spraying coating bioactive ceramics onto silicon nitride surface as composite endosteal implants. AB - The successful key of endosteal implants depends on the properties of implant materials which are very important for oral implantology at the present. Because silicon nitride has high strength and hydroxylapatite (HA) and flourapatite (FA) have good biocompatibility. In this paper, we apply silicon nitride as base material. Plasma spray HA, FA onto its surface as composite endosteal implants. Physical and chemical properties test, includes X-ray diffraction (XRD), scanning electronic microscope (SEM), EDAX and bonding strength test (push-out test). The results indicate: after plasma-spraying coating, crystalline phase of HA and FA unchanged and form a lot of pores among the crystal particles. Those pores benefit bone growing into them. It is very important for implants to be fixed in bone for long time, Ca/P ratio has no significant change. Bonding strength test results indicate: Si3N4-HA 23.6MPa, Si3N4-FA 27.12 MPa are higher than that of Ti HA 15.07 MPa. On the basis of these studies, they are kinds of ideal implant materials. PMID- 9731427 TI - A new fuzzy logic based image enhancement. AB - In this paper a new fuzzifier (fh) and a new contrast intensification operator (NINT) have been proposed for the enhancement of computerized tomography (CT) images. The performance of the proposed technique has been compared with an existing fuzzy logic based enhancement technique in terms of a new fuzzy contrast measure. The superiority of the proposed technique has been established. PMID- 9731428 TI - [Comparison of the antioxidant effect of estriol and estradiol on low density lipoproteins in post-menopausal women]. AB - BACKGROUND: Estradiol (E2) has a potent antioxidant effect on low density lipoproteins (LDL) in vitro and in vivo, which could be important in explaining the cardioprotective effect of hormone replacement therapy (HRT) in post menopausal women. Estriol (E3), on the other hand, is a weak estrogen with low metabolic effects on different tissues, and at present no cardioprotective effect has been attributed to this steroid. AIM: To study the antioxidant effect of E3 on LDL and to compare it with the potent antioxidant action exhibited by E2. SUBJECTS AND METHODS: After LDL was isolated by ultra centrifugation from plasma of 12 healthy untreated post menopausal women, it was divided into aliquots containing 0.5 mg of LDL protein. Estriol and E2 in doses of 0, 1, 5, 15 and 50 microM were incubated with different aliquots of LDL. CuSO4 15 microM was added to each aliquot to induce an oxidative stress. The aliquots were then incubated during 4 hours at 37 degrees C. Malonaldehyde (MDA) was measured as a marker of LDL oxidation, and expressed as nM/mg protein. RESULTS: (mean +/- SD): Estriol induced a dose-dependent decrease in MDA concentration (baseline 62.8 +/- 21.7; 1 microM: 61.5 +/- 23.0; 5 microM: 52.9 +/- 20.3; 15 microM 43.5 +/- 20.1 and 50 microM: 31.0 +/- 17.6 nM/mg protein; F = 92.4; p < 0.0001), reaching a mean decrease of 50.7% at the highest dose tested. Estradiol has a similar dose dependent decrease in MDA concentration (F = 60.2; p < 0.0001), revealing a more potent effect than E3 (p < 0.05), with a mean decrease of 67.4% at the highest dose tested. CONCLUSIONS: Our results demonstrate that estriol shows an important antioxidant action of LDL in vitro, although its effect is less potent than estradiol. These results raise the possibility that estriol could have a cardioprotective effect in post menopausal women, possibility that has not been yet demonstrated. PMID- 9731430 TI - [Predictive value of the ergometric test stress parameters for myocardial ischemia according to SPECT thallium-201 scintigraphy]. AB - BACKGROUND: Exercise EKG is used as the test of choice in the diagnosis of coronary artery disease. Classical parameters are angor and ST depression representing myocardial ischemia. AIM: To correlate exercise EKG parameters with SPECT 201Thallium to know their likelihood ratios for ischemia. PATIENTS AND METHODS: Two hundred seventy four patients (171 men), aged 58 years old as a mean, were studied. Of these, 23% had a prior myocardial infarction. The likelihood ratios for the presence of ischemia of ST depression, failure to increase blood systolic pressure, the presence of angor and its duration during stress testing were calculated according to the results of SPECT 201Thallium. Seventy one patients were also subjected to a coronary angiography. RESULTS: Among men, likelihood ratios for the presence of angor, failure to increase systolic pressure, ST alterations and duration of angor were 6.9, 6.15, 1.77 and 1.27 respectively. Among women, the figures were 5.45, 1.77, 0.58 and 1.4 respectively. The diagnostic accuracy of SPECT 201Thallium, when correlated with the results of coronary angiography, was 85%. CONCLUSIONS: Among men, the best exercise EKG predictors for myocardial ischemia were the failure to increase systolic blood pressure and the presence of angor. Among women the only significant predictor was the presence of angor. PMID- 9731429 TI - [Nutritional features of elders living in poor communities of the Metropolitan Region of Chile]. AB - BACKGROUND: The higher life expectancy of the Chilean population has required a greater concern about health and nutrition of elderly people. AIM: To assess the alimentation of people over 65 years old, living in communities of the Metropolitan Region with high social vulnerability. MATERIAL AND METHODS: A random and representative sample of 257 elders living in the poorest communities of santiago was studied. They were interviewed in their homes about feeding patterns, the alimentation of the day before and their nutritional status. Mean energy and nutrient consumption and their fitting to FAO/WHO energy and National Research Council micronutrient recommendations were calculated. A feeding quality index, that considers the adequacy of consumption of all nutrients, was calculated. RESULTS: Except for bread, median consumption of all groups of foods was below recommendations. Median energy and protein consumption was 1438 Kcal and 53.9 g in men and 1118 Kcal and 37.7 g in women. Adequacy of consumption of all nutrients was low and the intake of energy, protein, calcium, iron and folate was significantly lower in women than in men. The factors associated with feeding quality in these subjects were a last meal of the day before different from supper, more than 15 hours of morning fasting and sex. CONCLUSIONS: Alimentation of elders living in poor communities of Santiago is severely deficient. Focalized intervention models to improve this situation should be devised. PMID- 9731431 TI - [Reduced plasma volume and changes in vasoactive hormones in underweight pregnant women]. AB - BACKGROUND: Pregnant women with low weight/height (wt/ht) have lower plasma volume and reduced birth weight than women with normal wt/ht. AIM: To explore the hormonal mechanisms involved in these alterations. PATIENTS AND METHODS: Plasma volume, and several hormones related to plasma volume regulation were determined in 24 near term pregnant women with low wt/ht and in 30 with normal wt/ht. RESULTS: Newborns's weight, height and ponderal index were reduced in the low wt/ht group. Plasma volume (3042 +/- 101 vs 3631 +/- 101 ml, p < 0.001); plasma renin activity (7.5 +/- 0.9 vs 11.1 +/- 0.9 ng/ml/h, p < 0.01) and aldosterone (428 +/- 47 vs 710 +/- 58 pg/ml, p < 0.001) were significantly reduced in the low wt/ht group. Similar reductions were observed in serum estradiol and progesterone levels. Urinary kallikrein activity (354 +/- 112 vs 824 +/- 134 nmoles/24 h, p < 0.05), 6-keto-prostaglandin F1 alpha (561 +/- 90 vs 1121 +/- 165 ng/24 h, p < 0.05) and thromboxane B2 (110 +/- 29 vs 280 +/- 29 ng/24 h, p < 0.05) were also reduced in low wt/ht women. CONCLUSIONS: We postulate that the reduced levels of vasoactive hormones observed in pregnant women with low wt/ht may interfere with plasma volume expansion and, in turn, cause low birth weight. PMID- 9731432 TI - [Contribution of chromosomal abnormalities to in vitro fertilization failures]. AB - BACKGROUND: Present knowledge of mechanisms involved in human fertilization has uncovered a new group of pathologic conditions that have been generically named fertilization abnormalities. AIM: To determine the contribution of chromosomal alterations to in vitro fertilization failures. MATERIALS AND METHODS: A cytogenetic analysis of oocytes that were not fertilized after insemination with normal spermatozoa. Oocytes were obtained from patients subjected to in vitro fertilization in a public hospital of Metropolitan Santiago. Ovulation was induced in these patients administering GnRh-a, FSH, HMG and HCG. The double fixation technique described by Wramsby was used to obtain chromosomes. RESULTS: One hundred and seven oocytes coming from 45 women aged 25 to 42 years old were studied. The frequency of aneuploidy in these oocytes was 37.3%, with a 11.8% of hypohaploidy, a 21.6% of hyperhaploidy and a 3.9% of diploid oocytes. In hyperhaploid as well as in hypohaploid oocytes, the chromosomes involved in aneuploidy pertained to groups D. and G. CONCLUSIONS: Although the total frequency of aneuploidy is within normal ranges, the frequency of hyperhaploidy is superior to previous reports. An explanation for this finding could be that the occurrence of a lack of disjunction with chromosomal retention in the parental cell occurs with a higher frequency than that in which the chromosomes are retained in the polocyte. We also suggest that oocyte chromosomal aneuploidy could contribute to the failure of in vitro fertilization procedures. PMID- 9731433 TI - [Effect of casein derived peptides on D-xylose absorption and small intestinal motility in dogs]. AB - BACKGROUND: The presence of food in the intestinal lumen increases absorption from an isolated intestinal loop, the mechanisms involved are unknown. Casein, and its respective hydrolysate, increased D-xylose absorption in both normal volunteers and experimental animals; this effect was associated with prolonged small intestinal transit time and a decrease of motor activity. AIMS: To separate from casein hydrolysate, groups of peptides and to investigate their effects on both D-xylose absorption and small intestinal motility. MATERIAL AND METHODS: Studies were performed on five dogs with a surgically implanted duodenal cannula. Three groups of peptides were separated by means of a Silica Gel 60 column and were continuously infused through the duodenal cannula. After 15 min, 5 g of D xylose were injected in the duodenum, plasma levels were measured, and the area under the curve was estimated. Motility was recorded by means of infused catheters and external transducers. RESULTS: Plasma levels of D-xylose were significantly increased during the infusion of one group of peptides compared to the others. In addition, the area under the curve: 3366 +/- 885 mg x min-1 observed with this group was significantly greater than the other two groups: 1432 +/- 183 mg x min-1 and 1137 +/- 280 mg x min-1 respectively. No statistically significant differences in motor activity were observed between the different groups of peptides. CONCLUSIONS: A group of peptides derived from casein was characterized by increasing D-xylose absorption. The presence of beta casomorphines might be the possible mechanism involved. PMID- 9731434 TI - [Mutations of the p53 suppressor gene in gastric adenocarcinoma]. AB - BACKGROUND: We have shown numeric alterations such as hyperploidy and hypoploidy with loss of chromosome 17 in primary gastric cancer. This chromosome maps p53 suppressor gene that induces the transcription of genes related to cellular cycle control, DNA synthesis and repair, cellular differentiation and apoptosis. AIM: To analyze, at a molecular level, the possible alterations of p53 suppressor gene in samples of gastric cancer and non tumoral mucosa. MATERIAL AND METHODS: Tissue samples of gastric carcinoma and non tumoral gastric mucosa coming from 26 patients subjected to a total gastrectomy were analyzed. The mutation of p53 suppressor gene exons 7 to 9 were determined using a conformational polymorphism analysis in single strands of the gene and indirect sequencing in some cases. RESULTS: Alterations in p53 gene were found in 77% of tumoral and 19% of non tumoral samples. T insertions in codons 260, 317 and 321, G insertion in codon 328 and G-T transvertion in codon 302 were found. Aminoacid sequence analysis of p53 protein obtained with sequencing data showed that T insertion in codon 260 could translate three erroneous aminoacids after the mutation and produce a truncated protein due to the creation of a stop codon. No associations between alterations of p53 gene and clinical or pathological variables such as age, sex, tumor localization, histological type and presence of lymph node metastases were observed. CONCLUSIONS: Mutations of p53 suppressor gene are frequent in gastric carcinoma. PMID- 9731435 TI - [Correlation between the presence of cytomegalovirus antibodies and antigen in blood leukocytes for the diagnosis of primary active infection]. AB - BACKGROUND: The diagnosis of cytomegalovirus infections measuring IgG or IgM antibodies has a high rate of false positive or negative results, specially in immunocompromised patients. AIM: To compare the diagnostic yield of antibodies against cytomegalovirus with the measurement of the antigen in peripheral leukocytes. MATERIAL AND METHODS: Forty three blood samples coming from pediatric patients with suspected cytomegalovirus infections were analyzed. Low affinity IgG and IgM antibodies against Epstein Barr virus and cytomegalovirus, using indirect ELISA assays, and the virus antigen in peripheral leukocytes, using a commercial immunoperoxidase assay, were measured. RESULTS: Seven patients had positive IgM antibodies against cytomegalovirus. In five of these the viral antigen was detected in peripheral leukocytes. Twenty patients had positive antibodies against Epstein Barr virus, and in 16 patients all serologic tests were negative. CONCLUSIONS: There is not a good correlation between antibodies against cytomegalovirus and the detection of its antigen in patients with acute infections. PMID- 9731436 TI - [Devices for saliva collection from the major salivary glands. Results in normal subjects]. AB - BACKGROUND: Collection of saliva produced by the major salivary glands may be accomplished either by cannulation of the glandular ducts or by the application of specific collecting devices to the emergence area of the glandular ducts. Those procedures are complex, slow, invasive and require skilled personnel. AIM: To report the design and application of a device to collect parotid saliva (snail collector) and another device to collect saliva from the submandibular/sublingual complex. MATERIAL AND METHODS: The saliva collection devices were tested in 40 healthy volunteers (20 male) aged 18 to 22 years old. Saliva was collected using conventional conditions, during 5 to 15 min. RESULTS: An average of 1 to 1.5 ml of saliva was collected in the 10-15 min period from both parotid and submandibular/sublingual glands. Flow rates from parotid glands were 80 microliters/min and 180 microliters/min from submandibular/sublingual glands. Parotid saliva had a protein and organic material concentration twice as high than saliva from submandibular/sublingual glands. The presence of human alpha amylase duplet (Mr 55 kD and 58 kD) predominated in parotid saliva, whereas saliva from submandibular/sublingual glands had other molecular markers such as the lysozyme duplet (Mr 18.5 kD and 17 kD). CONCLUSIONS: The tested devices were easily applicable, comfortable and allowed the collection of both parotid saliva and submandibular/sublingual saliva from various subjects at once, under the supervision of a single professional. PMID- 9731437 TI - [Cholestatic hepatitis associated with piroxicam use. Case report]. AB - Most nonsteroidal antiinflammatory drugs can produce hepatotoxicity. We report a 22 years old female who presented with an acute cholestatic hepatitis after a prolonged period of piroxicam use. Hepatitis was attributed to this drug since all markers for hepatitis virus (A, B, C, E, Epstein Barr, Cytomegalovirus and Herpex Simplex) were negative, autoimmune markers were negative, serum iron and ceruloplasmin were normal, there was a temporal relationship between the administration of piroxicam and the hepatitis, the histological picture was compatible with this etiology and the patient had a favorable evolution after the discontinuance of the drug. This type of hepatotoxicity is not common but it must be born in mind when patients must receive nonsteroidal antiinflammatory drugs for prolonged periods. PMID- 9731438 TI - [Merkel cell carcinoma. Report of 4 cases]. AB - We report four cases of Merkel cell carcinoma, a rare cutaneous neuroendocrine neoplasm with a high malignant potential. The first patient is a 90 years old male presenting with a tumor in the left superior eyelid. Surgical excision was the only treatment and seven months later, a local and regional tumoral relapse caused the death of this patient. The second patient is a 83 years old female with a tumor in her left ear. She was treated surgically and with radiotherapy, being free of disease after five years of follow up. The third patient is a 45 years old male with a tumor in the left forearm. Treatment was surgical excision of the primary tumor and axillary lymph nodes. He received post operative radiotherapy and is free of disease after three years of follow up. The last case is a 76 years old male, who was subjected only to an excisional biopsy of a lesion located in the left knee. He had a tumor relapse with inguinal and crural lymph node involvement that caused his death 12 months later. PMID- 9731439 TI - [Cyclosporosis. Report of a clinical case in Concepcion, Chile]. AB - Cyclospora cayetanensis is a newly recognized parasite widely distributed throughout the world, and isolated from children, immunocompetent adults and HIV infected individuals. The clinical manifestations of the infection are watery prolonged diarrhea, anorexia, fatigue, nausea, bloating and weight loss. In immunocompetent individuals, diarrhea is usually self limited but may last several weeks. In immunocompromised hosts it is prolonged, severe and can be associated with biliary tract involvement. We report a 50 years old female that, five days after returning from Cuba, presented with low grade fever, anorexia, fatigue, explosive diarrhea and weight loss. Physical examination was normal. A stool specimen contained many organisms with morphological features of Cyclospora caetanensis. The diagnosis was confirmed at the Centers for Disease Control (Atlanta Ga). PMID- 9731440 TI - [Inspiratory muscle training in patients with chronic obstructive pulmonary disease]. AB - We analyze the effect of inspiratory muscle training (IMT) in patients with chronic obstructive pulmonary disease (COPD), with special emphasis on its effects on inspiratory muscle function and clinical outcomes. We reviewed only randomized, controlled studies that have either controlled both the load and the breathing pattern when using resistive training or have employed a threshold trainer in which the load is independent of the pattern of breathing, since methodological aspects may explain inconsistent results in the literature. In these circumstances, most of the studies demonstrated positive effects on inspiratory muscle function. Clinical effects were seldom evaluated; limited available data showed a reduction in dyspnea that was related to an increase in maximal inspiratory pressures (PIMax). When exercise capacity was evaluated through the distance the patients were able to walk in 6 or 12 minutes, most studies demonstrated a significant increase. Other reported positive effects were improvement in nocturnal SaO2, inspiratory muscle power output and maximal inspiratory flow rate. Based in this review, a recommended training regime appears to be an intermediate load (30-40% PIMax) using a threshold device for 30 minutes daily for at least 5 weeks. Although in the literature the criteria for selecting patients are not always well defined, we consider IMT as a helpful procedure for pulmonar rehabilitation in those patients with a moderately severe inspiratory muscle dysfunction presenting dyspnea during daily living activities despite optimal therapy. PMID- 9731441 TI - [Diagnosis disclosure: the place of beneficence, autonomy and veracity in medical ethics]. AB - OBJECTIVE: The moral doctrine of diagnosis disclosure is derived from a respect for the patient's autonomy as well as the patient's beneficence. These two goals are not necessarily incompatible, but they often lead to different decisions about what information needs to be shared with patients. The ethics of veracity does not prescribe duties, either old or new, but implies a new way of thinking about ethics. METHOD: Theories of the three general sorts just mentioned are identified and explained here. RESULTS: The ethics of veracity as such is limited to unmasking the falsifications of desire that inhabit the moral life. This would meet the moral goals of shared decision-making, because patients could make sense of such data. CONCLUSIONS: Certainly other moral doctrines may turn out to be relevant to the problems discussed in this paper. Nonetheless, they are sufficiently comprehensive to provide an analytical framework by means of which moral problems of diagnosis disclosure can be evaluated. We cannot expect anything else from the enterprise of the ethics of veracity but a critique of authenticity. Its force is that of suspicion, not justification or legitimation, and still less that of prescription. PMID- 9731442 TI - [Antiretroviral therapy. Announcement of the Advisory Committee of the Chilean Society of Infectology on AIDS]. AB - There are treatments to induce the long term suppression of viral replication and that delays the progression of HIV disease. To be effective, these treatments require the uninterrupted use of a combination of drugs (ideally three), patients must be highly compliant, must be instituted in early stages of the disease and drugs must be used for prolonged periods and given by specialists. These treatments are indicated in all symptomatic patients and in those with early immunologic deterioration or high viral load. Recent infection and acute primary retroviral infections should also be considered for treatment. The treatment sponsored and financed by the ministry of health for its beneficiaries is insufficient at this time, since it is received by a minority of eligible patients due to budgetary reasons, and only two drugs are given which is considered such optimal by most experts. The committee considers that the responsibility for financing, providing and delivering these treatments in proper combination and dosages exceeds the duty of the Ministry of Health and should include all the involved parties. However, the state and its official institutions have a special responsibility to provide with adequate treatments to the poorer segments of our population. They also should promote, supervise and control the proper access of the rest of the population to efficient treatments. The committee also considers that the efforts to prevent new infections must not be neglected and that individuals under successful treatment should not consider themselves as non infectious. PMID- 9731443 TI - [Common mental disorders, depression and public health]. AB - Common Mental Disorders (CMD), particularly depressive disorders, are problems of public health importance and it would seem appropriate to approach them from a public health point of view. In this paper only psychosocial aspects are dealt with, because biological factors are less important in the pathogenesis of CMD. Although there are significant associations between socioeconomic variables and CMD, the association between poverty and CMD seems to be one of the most robust. There is good evidence that low income is associated with CMD, and this relationship may be causal. There is therefore a possibility that changes in social and economic policy that would reduce poverty might also reduce the prevalence of CMD. Future research should try to unravel the mechanisms and causal factors that underlie these common and disabling disorders. PMID- 9731444 TI - [Change in attitude to the management, diagnosis and therapy of sleep apnea syndrome. Hungarian Health Service]. AB - Sleep apnea syndrome occurs in 4% of the general population. Due to its incidence, clinical symptoms and its causal or precipitating roles in other diseases, sleep apnea represents a real public health problem. Its early identification and its adequate treatment require a continuously functioning system of screening and care based on the family doctors and the therapeutic centres. The efficacy of such a system consist of following factors: integrated action in screening, care, treatment, postgraduate and health education, suitable regional distribution of basic screening and specialized care and a new type of cooperation among specialists working together in specialized care. The aim of this paper is to present an effective system for its management relying on both the international know-how and the experience obtained in Hungary, which has been adjusted to the structure of the Hungarian Health Service. PMID- 9731445 TI - [The role of the histopathological analysis of sentinel lymph nodes in breast cancer. Preliminary findings]. AB - Axillary lymph node status is the single most powerful prognostic marker for breast cancer. Histopathological assessment of lymph nodes has become the gold standard, although conventional histological work-up may miss 10-20% of node positive cases, potentially resulting in undertreatment and poorer survival of these patients. Identification and detailed histological assessment of sentinel lymph nodes may improve the error rate of conventional methods. We performed the first 30 lymphatic mappings using patent blue vital staining at our department of Surgery in the second semester of 1997. The success rate of identifying 1 or 2 sentinel nodes was 73.3% (22 cases). Axillary dissection and either breast conserving surgery of mastectomy were performed on all patients. Sentinel lymph nodes were serially sectioned and also investigated by immunohistochemistry using primary antibodies to cytokeratin and epithelial membrane antigen. This correctly predicted the qualitative axillary nodal status gained from all the nodes in 21 cases (95.5%). The only false negative sentinel node was associated with a micrometastatis in a non-sentinel lymph node. From the predictive cases 10 (47.6%) had positive nodes, and half of these had metastases only in the sentinel node. To our knowledge, we are the first in Hungary to report preliminary results from a lymphatic mapping study for breast cancer. It seems evident that assessment of sentinel lymph nodes increases the sensitivity of the less reliable conventional histopathological work-up, and this provides a more accurate staging when performed in conjunction with axillary dissection. On the other hand negativity of the sentinel lymph node may question the need for the clearance procedure. PMID- 9731446 TI - [Diagnostic studies of cerebrospinal fluid in patients with multiple sclerosis]. AB - The diagnostic criteria postulated by Poser necessitate clinical and laboratory CSF analysis for establishment of the diagnosis of definitive multiple sclerosis. The present paper reports methods for CSF examinations relating to multiple sclerosis with regard to the examinations suggested by the Charcot Foundation. In the course of CSF analysis, it is important to discriminate between the immunoglobulins present in normal amounts, those synthesized locally in pathological quantities and those penetrating across the damaged blood-CSF barrier. Normally, a parallel assay of CSF and serum specimens is carried out in the course of quantitative and qualitative protein analysis. In 37 patients with clinical multiple sclerosis, we determined the albumin and the immunoglobulin classes IgG, IgA and IgM, using laser nephelometry. An elevated IgG index was found in 76% of the cases, which points to local IgG snythesis and might be proof of the humoral immune response. The albumin quotient, which is suitable for examination of the integrity of the blood-CSF barrier, was within the reference range. Qualitative protein analysis was performed by means of electrophoresis on agarose-gel and isoelectric focusing. Agarose-gel electrophoresis revealed oligoclonal gammopathy in 68%, in contrast with the 91% demonstrated by isoelectric focusing. Comparison of the two kids of qualitative protein analyses indicated that isoelectric focusing was more sensitive for the detection of oligoclonal bands, in support of the literature finding. PMID- 9731447 TI - [Chlamydia pneumoniae infection in patients with myocardial infarct and angina pectoris]. AB - In a prospective open study we investigated Chlamydia pneumoniae infections in 36 consecutively admitted patients: 26 males, mean age 53.4 yr, range 36-70 yr, 10 females mean age 57.7 yr, range 47-70 yr, suffering myocardial infarction (24 acute, 2 previous) or angina pectoris (10). Antibody serum levels were measured by the immunefluorescent method and they were as follows: negative 5, low 12, medium/high 11, chronic infection 5, recent infection 3. The 3 cases considered as recent infections are described in detail. PMID- 9731448 TI - [Kearns-Sayre syndrome]. AB - The authors describe a rare group of symptoms, resulting in progressive external ophthalmoplegia, retinal pigment epithelial dysfunction and cardiac conduction disturbance. The illness belongs to the group of mitochondrial cytopathies. The case extends over the diagnostic possibilities, with special attention on electromyographic diagnostic, clinical symptoms, pathomechanism of the disease, and the therapic possibilities. PMID- 9731449 TI - [Istvan Apathy and neurofibrils]. PMID- 9731450 TI - Character is fate: the life of Paul B. Beeson. PMID- 9731451 TI - Charles Dickens gets a lesson in sensitivity: Mrs. Jane Seymour Hill's reaction to David Copperfield. PMID- 9731452 TI - On interns and interning. PMID- 9731453 TI - Reacting to epidemics--past and present. PMID- 9731454 TI - Angelika: the impact we have as physicians. PMID- 9731456 TI - The public stake in physician authority. PMID- 9731455 TI - Blalock and Harrison--a rare friendship. PMID- 9731457 TI - Physician-assisted suicide. PMID- 9731458 TI - Good clinical teachers. PMID- 9731459 TI - Commitment and charisma. PMID- 9731460 TI - Futures market in organ procurement. PMID- 9731461 TI - Where is the sacrifice? Drs. Soderdahl reply to Dr. Atkins (Pharos, Spring 1998). PMID- 9731462 TI - Abandonment of rotating internships. PMID- 9731464 TI - WHO Expert Committee On Biological Standardization. AB - This report presents the recommendations of a WHO Expert Committee commissioned to coordinate activities leading to the adoption of international requirements for the production and control of vaccines and other biologicals and the establishment of international biological reference materials. The report starts with a discussion of general issues brought to the Committee's attention and provides information on the status and development of reference materials for various antibiotics, antibodies, blood products, cytokines, endocrinological and related substances, toxins and other substances. The second part of the report, of particular relevance to manufacturers and national control authorities, contains requirements for the use of animal cells as in vitro substrates for the production of biologicals, guidelines for the production and control of the acellular pertussis component of monovalent or combined vaccines, as well as guidelines for assuring the quality of DNA vaccines. PMID- 9731463 TI - George Thorn and Cushing's. PMID- 9731465 TI - Retroviral vectors for gene therapy. AB - Although there are many hurdles to overcome for successful gene therapy, there is a vast potential to permanently incorporate genes into cells to correct genetic disorders and to combat viral infections. Retroviruses, inspite of some limitations, offer the best hope in this direction and lentiviral vectors, which infect nondividing cells, may be the choice in the future, especially in gene therapy for central nervous system disorders. PMID- 9731466 TI - Detection of plant viruses--biotechnological and molecular advances. AB - Recent advances in biotechnology and molecular biology have played a significant role in development of rapid, specific and sensitive assays for detection of plant viruses. Production of monospecific polyclonal antibodies, monoclonal antibodies have enabled to group isolates of viruses and distinction of closely related strains. In cDNA hybridization applications, there is an increasing interest to employ non-radioactive probes for detection of nucleic acids. Detection limit of nucleic acid is remarkably comparable to those of radioactive labelled probes. Application of polymerase chain reaction (PCR) has made it possible to amplify the low numbers of viral RNA/DNA molecules and their subsequent detection. Underlying principles, their advantages and disadvantages for application of monospecific polyclonal antibodies, hybridoma technology, molecular hybridization and PCR technology with reference to detection of plant viruses have been discussed in this review. PMID- 9731467 TI - Sequence analysis of full length cDNA for enhancing factor/phospholipase A2. AB - Enhancing factor (EF) protein was initially purified as a modulator of epidermal growth factor from small intestines of mouse. The cDNA sequence, obtained by RT PCR, revealed that EF belonged to the non-pancreatic, phospholipase A2 (PLA2) family. This was the first report of the mouse PLA2. In the present paper we report the complete cDNA sequence of EF gene, in which the 5' sequence has been obtained by RAcE-PCR. The predicted amino acid sequence was computer analysed and the putative sites for enzyme action, calcium binding and heparin binding have been identified. The complete protein sequence of EF along with 16 aligned sequences were used to infer a phylogenetic tree. From this data the mouse EF was grouped with other membrane associated PLA2 with a bootstrap value of 98% indicating that it belonged to this class. PMID- 9731468 TI - Immunohistochemical co-expression of human papillomavirus type 16/18 transforming (E6) oncoprotein and p53 tumour suppressor gene proteins in oesophageal cancer. AB - Human papillomaviruses have been widely implicated as important etiologic agents in various squamous cell carcinomas including oesophageal carcinoma. p53 mutant oncoprotein has also been implicated in various tumours. Immunohistochemical analysis was employed to detect the co-expression of HPV and p53 mutant protein in biopsy specimens of patients of cancer oesophagus as well as controls. This analysis revealed a significantly higher immunopositivity (63%) of E6 oncoprotein of HPV 16/18 in carcinoma of the oesophagus. Immunoexpression of E6 oncoprotein of HPV did not alter significantly the degree of differentiation of the tumour. Seventy-seven percent of cases of oesophageal carcinoma showed strong immuno staining for mutant p53 protein. A higher percentage (89%) of tissues showed immunoexpression of mutant p53 protein in conjunction with E6 oncoprotein of HPV 16/18 indicating a selective degradation of key cellular protein of p53 having regulatory properties which in turn leads to uncontrolled cellular proliferation. Therefore, coexpression of oncoprotein E6 of HPV 16/18 and mutant p53 protein may be considered as a "high risk" factor for progression to oesophageal malignancy. PMID- 9731469 TI - Effect of estrogen and progesterone on newly synthesised and released human fallopian tube specific proteins. AB - Current study was carried out to identify the profile of newly synthesized and released proteins by human fallopian tube (hFT). Results indicated that hFT during menopause synthesised and released only 2-3 proteins as against several proteins ranging from molecular weight (MW) approximately 20 to approximately 130 kD during normal menstrual cycle. In vitro addition of estradiol-17 beta (E2) resulted in synthesis and release of a number of proteins including specific protein of MW 110-130 kD. Addition of progesterone (P) however, led to inhibition of protein synthesis and a combination of E2 and P negated the effect of the latter. An alteration in oviductal secretory protein-profile following addition of E2 in vitro were similar to that observed during normal menstrual cycle. PMID- 9731470 TI - Evaluation of topical formulations of aqueous extract of Centella asiatica on open wounds in rats. AB - Formulations (ointment, cream and gel) of aqueous extract of C. asiatica, when applied topically, thrice daily for 24 days on the open wounds in rats increased cellular proliferation and collagen synthesis at the wound site, as evidenced by increase in collagen content and tensile strength. The treated wounds epithelialised faster and the rate of wound contraction was higher as compared to control wounds. The process of healing was better with gel formulation when compared to other two formulations. PMID- 9731471 TI - Protective effect of turmeric (Curcuma longa L.) extract on carbon tetrachloride induced liver damage in rats. AB - The protective effect of tumeric extract (TE) in diet on CCl4-treated rats was studied. Rats were divided into 5 groups: (1) untreated, (2) CCl4 treated, (3) pre-TE for 2 weeks followed by CCl4, (4) TE + CCl4 given concurrently and (5) 5% TE as positive control. The serum levels of bilirubin, cholesterol, aspartate aminotransferase, (AST), alanine amino transferase (AST), (ALT) and alkaline phosphatase were estimated after 1, 2 and 3 months. CCl4 caused a maximum increase (2-3-fold in all the above parameters. As compared to CCl4 group, a short pre-treatment of TE showed reduction in cholesterol, bilirubin, AST, ALT and alkaline phosphatase activity whereas concurrent treatment of TE + CCl4 reduced to a greater extent the levels of all parameters except ALT. To conclude, concurrent treatment of TE gave significant protection against CCl4 though the values did not reach the normal levels. PMID- 9731472 TI - Effect of current on dermal permeation of positively charged liposomes. AB - Transdermal permeation of positively charged liposomally entrapped diphenhydramine hydrochloride (DPH-HCL) has been investigated in presence of pulse D.C. anodic current. Positively charged liposomes were prepared by lipid film hydration technique with stearyl amine as a charge inducer. The prepared liposomes were then subjected to in vitro permeation studies using artificial membrane (cellophane membrane) and human cadaver skin under the influence of iontophoretic current. The effect of variable current density as well as time frequency of application of current onto the release pattern of the plain drug and charged liposomally entrapped drug were studied and the results were compared. The results indicate that application of pulse D.C. anodic current significantly influences the transfer of positively charged liposomally entrapped DPH-HCL across HCS. PMID- 9731473 TI - Antitumour and radioprotective action of Podophyllum hexandrum. AB - A significant antitumour effect of P. hexandrum, a herb thriving at Himalayas (2500-4000 m), was observed in strain 'A' mice carrying solid tumours developed by transplanting Ehrlich ascites tumour (EAT). Subtoxic well tolerated sequential doses of aqueous extract of P. hexandrum (a daily dose of 34.5 mg/kg b.w. for 15 days) enhanced tumour doubling time (TDT) from 1.94 +/- 0.26 days to 19.1 +/- 2.5 days. However, no synergism was revealed between radiation and P. hexandrum, though both independently manifested antitumour effects. In normal mice, pre irradiation administration of extract of P. hexandrum protected mice in a dose dependent manner (optimal dose being 34.5 mg/kg body.wt. rendering 72% survival for 30 days) against whole body lethal irradiation of 10 Gy. Radioprotective properties of P. hexandrum were found to be comparable to synthetic radioprotectors like diltiazem etc. PMID- 9731474 TI - Effect of gamma irradiation on chemical and biological properties of lipopolysaccharide from Salmonella typhimurium. AB - Lipopolysaccharide (LPS) from S. typhimurium on exposure to gamma-radiation resulted in decrease in toxicity and was less mitogenic, Silver stained profiles of irradiated LPS on polyacrylamide gels revealed complete loss of its heteropolysaccharides which was confirmed further by analysing lipid A and LPS from Salmonella minnesota Re mutants on SDS-PAGE. Glucosamine and 2-keto 3-deoxy octonate(Kdo) contents were significantly decreased on treatment. Lipid A obtained by removal of heteropolysaccharides from LPS was less toxic on exposure to gamma radiations. PMID- 9731475 TI - Role of antioxidant defence in renal pathophysiology: primacy of glutathione peroxidase. AB - Antioxidant defences consisting of superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx), and non-protein thiol (GSH) were estimated in the surviving kidney of unilaterally nephrectomized rats on days 1, 2, 3 and 12, following surgery. Simultaneously renal tissue lipid peroxidation (LP) and plasma GPx was monitored. The aim of this study was to evaluate the role of antioxidant defence in renal pathophysiology. Significant elevations in renal LP, GPx and GSH were evident on the first day and were maintained till 3rd day following nephrectomy. The response of SOD and catalase was slower, attaining significant elevation on 2nd and 3rd day respectively. Tissue GPx was the only parameter which remained elevated till 12th day, while all the other parameters returned to control levels by day 12. Plasma GPx declined significantly on days 3 (13.2%) and 12 (19.6%). These changes appear to be a part of the process when functions of 2 kidneys are taken over by a single kidney following unilateral nephrectomy. PMID- 9731476 TI - Is TAME a potent constrictor of non-airway smooth muscles? AB - Under in vitro conditions N-alpha-tosyl L-arginine methyl ester (TAME) induced a concentration dependent contractile response on ileal strips with EC50 of 4.3 x 10(-5) M as compared to acetylcholine which induced sustainable contractions with EC50 of 3.2 x 10(-6) M. The present study is the first to demonstrate that TAME is a potent constrictor of non-airway smooth muscle. PMID- 9731477 TI - Coagglutination test: a simple and rapid immunodiagnostic test for parvovirus infection in dogs. AB - The coagglutination test (COAT) was developed and standardized to detect canine parvovirus (CPV) antigen in faeces of infected dogs. Anti-parvovirus serum was raised in dogs for coating protein-A containing Staphylococcus aureus Cowan I strain. Agglutination of antibody coated bacteria invariably occurred within 2-3 min when mixed with standard CPV antigen or faecal supernatants of dogs having 8 or more haemagglutination (HA) titre of parvovirus antigen. The test had a perfect correlation with HA test and was found to be slightly more sensitive than agar gel precipitation test (AGPT) in detecting CPV antigens. As COAT is easy and needs no specific equipment or much technical know how to perform, it can be used as a field test for rapid clinical diagnosis of parvovirus infection in dogs. PMID- 9731478 TI - A phytoalexin is modified to less fungitoxic substances by crown rot pathogen in groundnut Arachis hypogaea L. AB - Phytoalexins from four different treatments viz. control, AF 1-treated, A. niger treated, and dual inoculated were separated by TLC showed that one phytoalexin with an Rf value of 0.628 (P1) appeared on 2nd day only in dual-inoculated seeds of groundnut (A. hypogaea). By 3rd day three additional phytoalexins were visualized in response to A. niger-treatment with lower Rf values 0.485 (P2), 0.388 (P3) and 0.314 (P4) as compared to P1. In dual inoculated seedlings, P1 and P3 could be visualized while only P1 appeared in response to AF 1 on 3rd day. All the compounds lost fluorescence on exposure to light, got converted to pale yellow colour. In all the treatments no phytoalexin accumulation was observed after 3rd day. All the four phytoalexins had a major peak between 338 and 339.5 nm. In potato dextrose broth, the growth of A. niger showed a steady increase up to 32 hr while it was significantly inhibited with P1 in microtiter plates. P2, P3 and P4 (in the same order) had significantly less antifungal activity as compared to P1. The antifungal activity of the phytoalexins decreased with decrease in their Rf value. PMID- 9731479 TI - Increased expression of cyclooxygenase 2 occurs frequently in human lung cancers, specifically in adenocarcinomas. AB - Cyclooxygenase (COX)-2 expression was immunohistochemically examined in 59 human lung cancers as well as in normal and premalignant lung specimens. In contrast to scattered weak reactivity seen in normal peripheral airway epithelial cells, markedly up-regulated COX-2 expression was detected in about one-third of atypical adenomatous hyperplasias and carcinoma in situ specimens, and a significant increase in COX-2 expression was observed in 70% of invasive adenocarcinoma cases. Interestingly, the proportion of adenocarcinoma cells with marked COX-2 expression was much greater in lymph node metastases than in the corresponding primary tumors. In contrast, small cell carcinomas showed virtually negligible expression, and squamous cell carcinomas showed infrequent and low expression. These findings suggest that an increase in COX-2 expression may be associated with the development of adenocarcinomas and possibly with acquisition of an invasive and metastatic phenotype. PMID- 9731480 TI - Hypoxia and vascular endothelial growth factor expression in human squamous cell carcinomas using pimonidazole as a hypoxia marker. AB - Hypoxia in human tumors is associated with poor prognosis, but the molecular mechanisms underlying this association are poorly understood. One possibility is that hypoxia is linked to malignant progression through vascular endothelial growth factor (VEGF) induction and the associated angiogenesis and metastasis. The present clinical study measures hypoxia and VEGF expression on a cell-by-cell basis in human squamous cell carcinomas to test the hypothesis that hypoxia and VEGF protein expression are coupled in human tumors. Eighteen patients with invasive squamous cell carcinoma of the uterine cervix and head and neck have been investigated by a quantitative image analysis of immunostained sections from their tumors. The hypoxia marker pimonidazole was used to measure tumor hypoxia, and a commercially available antibody was used to measure VEGF protein expression. A quantitative immunohistochemical comparison of hypoxia and VEGF protein expression revealed no correlation between the two factors. PMID- 9731481 TI - The helix-loop-helix protein Id2 is overexpressed in human pancreatic cancer. AB - Id2 belongs to the Id family of helix-loop-helix (HLH) proteins, which upon heterodimerization with basic HLH proteins prevent basic HLH proteins from DNA binding. Proteins of the Id family act as negative regulatory transcriptional factors, and their expression correlates with cell proliferation and arrested differentiation in many cell lineages. In this study, we characterized the expression of Id2 in normal and cancerous pancreatic tissues. Pancreatic cancers markedly overexpressed Id2 mRNA in comparison to the normal pancreas. Furthermore, there was abundant Id2 immunoreactivity in the cancer cells within the pancreatic tumor mass. In PANC-1 human pancreatic cancer cells, steady-state Id2 mRNA levels increased upon serum addition and decreased after induction of differentiation with either sodium butyrate or 12-O-tetradecanoylphorbol-13 acetate. Inhibition of Id2 expression with Id2 antisense oligonucleotides inhibited the growth of these cells, whereas random and sense oligonucleotides were without effect. These findings suggest that Id2 may have a role in human pancreatic cancer. PMID- 9731482 TI - Antisense oligodeoxyribonucleotide against the MLL-LTG19 chimeric transcript inhibits cell growth and induces apoptosis in cells of an infantile leukemia cell line carrying the t(11;19) chromosomal translocation. AB - To clarify the role of the multiple lineage leukemia gene-leukemia translocation gene of chromosome 19 (MLL-LTG19) protein in leukemogenesis, we synthesized antisense oligodeoxyribonucleotide (ODN) against the fused region of the MLL LTG19 chimeric transcript and treated KOCL33 cells carrying the t(11;19) translocation with antisense ODN. The antisense ODN inhibited cell growth and induced apoptosis in KOCL33 cells but not in Daudi cells, which have no t(11;19). The levels of MLL-LTG19 mRNA and MLL-LTG19 protein in KOCL33 cells treated with antisense ODN were shown to decrease with time by reverse transcription-PCR and Western blot analysis. These results suggest that the MLL-LTG19 fusion protein contributes to cell proliferation and malignant transformation in infantile acute leukemia cells carrying the t(11;19) translocation. PMID- 9731483 TI - The coding region of the Bloom syndrome BLM gene and of the CBL proto-oncogene is mutated in genetically unstable sporadic gastrointestinal tumors. AB - Microsatellite instability (MSI) characterizes the hereditary nonpolyposis colorectal cancer syndrome but is also found in sporadic tumors. Frameshifts in microsatellites found in the coding regions (CDRs) of the TGFbeta1-RII, IGFIIR, hMSH3, hMSH6, and BAX genes indicate that MSI is involved in tumorigenesis by targeting genes that are directly implicated in the tumorigenic process. To identify additional genes targeted for MSI, we performed an analysis of the GenBank database that revealed 21 microsatellite repeats located in the CDR of 18 genes (12% of the analyzed sequences) whose function could be potentially associated with the tumorigenic process. Mutational studies of 57 sporadic gastrointestinal tumor DNAs revealed the presence of length variations in three of them: (a) BLM; (b) CBL; and (c) HOXA1. In the BLM gene, we found a frameshift mutation in a polyadenine repeat, whereas in the CBL proto-oncogene, an expansion of a trinucleotide repeat was detected with no translation shift. These alterations were present in 18 and 9%, respectively, of the genetically unstable sporadic gastrointestinal tumors analyzed, but in none of the cancers without the mutator phenotype. These changes were present in the DNA from the tumor but not in that from normal cells of the same patient. The HOXA1 retraction of a trinucleotide repeat was as frequent in both types of cancers and was also found in some normal paired tissues, therefore behaving as a neutral polymorphism. Our data extend the spectrum of unstable microsatellites located in gene CDRs and suggest that BLM and possibly CBL are involved in gastrointestinal tumorigenesis. Based on its proposed function, the BLM gene could represent a link between MSI and chromosomal instability pathways, because MSI targeting of the BLM gene could generate hypermutability and/or chromosomal instability. PMID- 9731484 TI - Topotecan lactone selectively binds to double- and single-stranded DNA in the absence of topoisomerase I. AB - We report the first experimental observation that a clinically important camptothecin [CPT; topotecan (TPT), a water-soluble CPT] binds directly and noncovalently to double-stranded DNA and single-stranded DNA structures in the absence of topoisomerase I, but only in the lactone form. We observed clear DNA sequence specificity of the TPT lactone binding to duplex DNA, which was comprised of alternating purine-pyrimidine sequences that contained dT. These structural studies of direct TPT lactone-DNA binding support several important considerations involving possible mechanism(s) of anticancer activity of CPT-type drugs containing a 20(S) lactone moiety. PMID- 9731485 TI - Frequent somatic mutations in serine/threonine kinase 11/Peutz-Jeghers syndrome gene in left-sided colon cancer. AB - We analyzed somatic mutation and loss of heterozygosity (LOH) in the serine/threonine kinase 11 (STK11)/Peutz-Jeghers syndrome gene in 49 colorectal tumors in three different stages of a dysplasia-carcinoma sequence. We detected LOH in 10 of 19 (52.6%) informative colorectal cancers at loci D19S886 and/or D19S883, but no LOH was observed in 25 informative adenomas. We detected a total of 9 somatic mutations [7 of 13 (53.8%) left-sided colon cancers and 2 of 7 (28.6%) left-sided adenomas with high-grade dysplasia], but no mutations were detected in right-sided colon tumors. Of the nine mutations, one was a frameshift mutation (the same mutation detected in Peutz-Jeghers syndrome family previously), and the other eight were missense mutations. This results indicate that STK11 is a tumor suppressor gene and that genetic changes of STK11 play an important role in left-sided colon cancer carcinogenesis. PMID- 9731486 TI - Quantitative 1H nuclear magnetic resonance diffusion spectroscopy of BT4C rat glioma during thymidine kinase-mediated gene therapy in vivo: identification of apoptotic response. AB - We have investigated the effects of thymidine kinase-mediated gene therapy in a malignant rat BT4C glioma by using 1H nuclear magnetic resonance spectroscopy in vivo. Ganciclovir has been successfully used in thymidine kinase gene therapy as treatment for various experimental malignancies. The cell damaging effect seems to be mediated by apoptosis, optimally leading to eradication of tumor tissue. In this study, we show that ganciclovir treatment of tumors transfected with the herpes simplex thymidine kinase gene causes profound changes in water, metabolites, and macromolecules observable by diffusion spectroscopy. During treatment, a 50% reduction from 0.14 +/- 0.01 x 10(-9) m2/s in the apparent diffusion coefficient of choline-containing compounds can be observed, concomitant with a 219% increase in the apparent diffusion coefficient of the rapidly diffusing water component. These changes are associated with an increase in the relative fraction of this water component from 87 to 94%. The apparent diffusion coefficients of the slowly diffusing water component and macromolecules remain unaltered. The results imply a reduction in cell size and number, a significant increase in intracellular viscosity, and a possible reduction in the hydrodynamic radii of macromolecular components, which are ascribed as biophysical signatures for apoptotic cell death. PMID- 9731487 TI - Cancer-associated expression of glycolipid sulfotransferase gene in human renal cell carcinoma cells. AB - Human renal cell carcinoma (RCC) tissue and a cell line derived therefrom, SMKT R3, showed markedly increased glycolipid sulfotransferase [cerebroside sulfotransferase (CST); EC 2.8.2.11] activity and accumulated sulfoglycolipids. Recently, we cloned a human CST cDNA from a SMKT-R3 cDNA library (K. Honke et al., J. Biol. Chem., 272: 4864-4868, 1997). In this study, we investigated the expression of the CST gene in seven human RCC lines (SMKT-R1, SMKT-R2, SMKT-R3, SMKT-R4, TOS-1, TOS-2, and ACHN) and their normal counterpart, human renal proximal tubular cells. On Northern blot analysis, a marked increase of CST mRNA was observed in every RCC line, except for ACHN, as compared with normal cells. ACHN cells showed a slightly increased level of CST mRNA. CST activity was correlated with the amount of mRNA. Sulfoglycolipid analysis revealed that expression of lactosylceramide sulfate was correlated with the CST level. Furthermore, we examined the effects of epidermal growth factor (EGF), tetradecanoylphorbol-13-acetate, and genistein, which are known to regulate CST activity in SMKT-R3 cells, on CST-gene expression in various RCC cells. On treatment with EGF, CST mRNA time-dependently increased in accord with its activity in SMKT-R3 cells. Yet, augmentation by EGF was only observed in SMKT-R3. In contrast, a reduction of CST mRNA and activity by tetradecanoylphorbol-13 acetate and genistein was observed in all of the lines examined. Taken together, these findings indicate that in human RCC cells, the CST gene is generally overexpressed via a signaling pathway involving protein kinase-C and tyrosine kinases. PMID- 9731488 TI - High susceptibility of p53(+/-) knockout mice in N-butyl-N-(4 hydroxybutyl)nitrosamine urinary bladder carcinogenesis and lack of frequent mutation in residual allele. AB - The loss of p53 functions is considered to compromise the growth-suppression machinery of the cell and facilitate neoplastic change. In humans, genetic alteration in the p53 gene is one of the most frequently observed molecular changes in tumors, including urinary bladder carcinomas. We have investigated the susceptibility of heterozygote p53 knockout mice to N-butyl-N-(4 hydroxybutyl)nitrosamine (BBN) in terms of urinary bladder tumor induction. Both p53(+/-) knockout mice and C57BL/6 original parent strain were administered 0, 0.002, 0.004, 0.0075 and 0.025% BBN in the drinking water for 20 weeks. As compared with the C57BL/6 strain, greater lesion yields were observed in knockout mice after 20 weeks of treatment. Transitional cell carcinomas were found in 9 (75%) and 12 (100%) of each 12 mice of the 0.0075 and 0.025% BBN treatment groups, respectively, whereas only 1 (11%) and 6 (67%) of each 9 of the C57BL/6 mice demonstrated tumors. Preneoplastic lesions (dysplasia) were also observed more frequently in the lower dose groups in the knockout mice than C57BL/6 mice. PCR single-strand conformation polymorphism analysis followed by DNA direct sequencing of the p53 gene (exons 5-8) extracted from bladder tumors demonstrated mutations in 3 of 11 (27.3%; exon 7) and 8 of 29 (27.6%; exons 5-8) tumors in C57BL/6 and knockout mice, respectively. There was no significant difference in the mutation rates at the residual p53 gene between the two cases. All mutations observed in knockout mice were restricted to the normal allele, and none were present in the gene-targeted null allele. In a separate experiment, 5-bromo-2' deoxyuridine labeling indices after treatment with BBN for 2 or 4 weeks were significantly higher in knockout mice than wild-type mice. Measurement of the urinary concentration of N-butyl-N-(3-carboxypropyl)nitrosamine, a proximate carcinogenic metabolite, revealed no significant differences between knockout and original parent strain after administration of 0.0075% BBN in the drinking water for 4 weeks. In conclusion, knockout mice are distinctly more sensitive to urinary bladder carcinogenesis induced by BBN than their original parent strain, as evidenced by elevated DNA synthesis during carcinogen administration and an increased tumor yield. The high susceptibility of p53 knockout mice appeared to be related to the high level of cell proliferation rather than that of N-butyl-N (3-carboxypropyl)nitrosamine in the urine or that of mutations at the p53 gene. PMID- 9731489 TI - Telomerase activity exclusively in cervical carcinomas and a subset of cervical intraepithelial neoplasia grade III lesions: strong association with elevated messenger RNA levels of its catalytic subunit and high-risk human papillomavirus DNA. AB - In this study, we investigated telomerase activity and human telomerase reverse transcriptase (hTERT) mRNA expression in relation to high-risk human papillomavirus (HPV) DNA presence in the spectrum of cervical premalignant lesions. Reconstruction experiments revealed that telomerase activity determined by the telomeric repeat amplification protocol assay and hTERT mRNA by reverse transcriptase-PCR could be detected in down to 100 and 1 SiHa cervical cancer cells, respectively. Telomeric repeat amplification protocol analysis on cervical tissue specimens revealed that none of the histomorphologically normal cervical samples (n = 8) and cervical intraepithelial neoplasia (CIN) grade I (n = 10) and grade II (n = 8) lesions had detectable telomerase activity. However, telomerase activity was shown in 40% of CIN grade III lesions (n = 15) and 96% of squamous cell carcinomas (n = 24). Despite the fact that hTERT mRNA was found at much higher frequencies, semiquantitative reverse transcriptase-PCR revealed that elevated hTERT mRNA levels were strongly correlated with detectable telomerase activity. Furthermore, telomerase activity and elevated hTERT mRNA levels were only detected in cases that contained high-risk HPV DNA. In contrast, low or undetectable hTERT mRNA levels were demonstrated in both high-risk HPV positive and negative cases. These data indicate that telomerase activity detectable with the assay used and concomitant elevated levels of hTERT mRNA reflect a rather late step in the CIN to squamous cell carcinoma sequence, which follows infection with high-risk HPV. PMID- 9731490 TI - Ionizing radiation inhibits chemotherapy-induced apoptosis in cultured glioma cells: implications for combined modality therapy. AB - Surgical resection followed by radiation therapy is the mainstay of treatment for glioblastoma multiforme (GBM), the most aggressive of the malignant gliomas. The poor clinical response of GBM and the intrinsic radiation resistance of this tumor type have prompted clinical investigations seeking to define the role of chemotherapy in the treatment of GBM. In this study, we examined the cytotoxic response of GBM-derived cell lines to treatment with both radiation and chemotherapy. We observed that the sensitivity of glioma cells to cisplatin- and FAS-induced apoptosis was diminished by prior treatment with ionizing radiation. Radiation conferred resistance to cisplatin and FAS cytotoxicity in a dose- and time-dependent manner. Radiation diminished the cisplatin-induced cytotoxicity of malignant glioma cells but failed to alter the cisplatin susceptibility of normal primary human astrocytes. Given the role of p53 in the response of cells to irradiation, we evaluated whether p53 function affects the observed radiation induced resistance to cisplatin. By examining isogenic cell lines differing only in p53 function, we demonstrated that radiation conferred resistance to cisplatin independently of p53. Current clinical strategies in the treatment of astrocytic tumors, which include combined modality therapy, have been empirically derived from limited clinical experience. Further understanding of the molecular determinants of apoptosis associated with combined modality therapy may guide the design of more efficacious multimodality protocols. PMID- 9731491 TI - Effects of some novel inhibitors of C17,20-lyase and 5alpha-reductase in vitro and in vivo and their potential role in the treatment of prostate cancer. AB - The effects of some novel steroidal compounds were evaluated against both human C17,20-lyase and 5alpha-reductase in vitro and also against androgen synthesis in normal male rats. L-2, L-36, L-37, and I-41 showed potent inhibition of human testicular C17,20-lyase, with IC50s of 43, 39, 42, and 58 nM, respectively. In contrast, ketoconazole, a competitive inhibitor of C17,20-lyase, had an IC50 of 76 n.M. L-36 also showed potent inhibitory activity against 5a-reductase in human prostatic microsomes, with an IC50 of approximately 31 nM. The inhibitory activities of L-2 and 1-41 on 5alpha-reductase were moderate, with IC50s of 75 and 151 nM, respectively, whereas L-37 showed little inhibitory activity against this enzyme. In comparison, finasteride, a potent inhibitor of 5alpha-reductase, had an IC50 of 33 nM. When normal male rats were treated with these novel compounds (50 or 100 mg/kg/day) for 14 consecutive days, the wet weight of the prostate was significantly reduced by L-36, L-37, and I-41, compared to the control group. Testosterone levels in rat serum were also reduced by L-36 (55%), L-37 (86%), and I-41 (53%). The concentrations of testosterone in rat testes were reduced by these novel compounds by 13-74%. The compounds also reduced the concentration of testosterone in rat prostates by 35-75%. Similarly, dihydrotestosterone (DHT) concentration in rat serum was reduced 30-89% by these compounds, compared to the control group. Prostatic DHT levels were also lower in rats treated with L-36 (48%), L-37 (54%), or I-41 (26%). In contrast, L-2 enhanced serum testosterone and prostatic DHT concentrations by >50%. These findings suggest that the dual activities of several of these novel inhibitors of C17,20-lyase and 5alpha-reductase accounts for the diminished levels of circulating androgens in vivo. PMID- 9731493 TI - Risk of anal carcinoma in situ in relation to human papillomavirus type 16 variants. AB - Infection with human papillomavirus (HPV), especially HPV16, is central to the development of squamous anogenital cancers and their precursor lesions, termed "squamous intraepithelial neoplasias." Men who have sex with men, particularly those who are infected with HIV, are at a high risk for anal infection with HPV16 and for low-grade anal neoplasia; however, only a subset of these men develop anal invasive cancer or its immediate precursor lesion, anal carcinoma in situ (CIS). To examine the hypothesis that certain variants of HPV16 are most strongly associated with development of anal CIS, we followed 589 men who have sex with men whose initial anal cytological smears did not show anal CIS. Anoscopy, anal cytology, and PCR-based assays for detection and classification of HPV types were performed every 4-6 months, with HPV16 further classified by single-stranded conformation polymorphism analysis as being a prototype-like (PL) or non prototype-like (NPL) variant. Anal CIS was histologically confirmed in 6 of 384 (1.6%) consistently HPV16-negative men, in 12 of 183 (6.6%) men with HPV16 PL variants, and in 4 of 22 (18.2%) men with HPV16 NPL variants. After adjustment for anal cytological diagnoses at study entry, HIV status and CD4 count, and detection of HPV types other than type 16, men with HPV16 NPL variants were 3.2 times (95% confidence interval, 1.0-10.3) more likely to develop anal CIS than were those with PL variants. Neither detection of HPV16 DNA at high levels nor detection of HPV16 DNA for a prolonged period, factors that we previously demonstrated to be associated with risk of high-grade anal squamous intraepithelial neoplasia, was significantly associated with HPV16 NPL variants. The biological mechanism relating to Ihis excess risk remains undetermined. PMID- 9731492 TI - Estrogenic effects of genistein on the growth of estrogen receptor-positive human breast cancer (MCF-7) cells in vitro and in vivo. AB - Genistein, found in soy products, is a phytochemical with several biological activities. In the current study, our research focused on the estrogenic and proliferation-inducing activity of genistein. We have demonstrated that genistein enhanced the proliferation of estrogen-dependent human breast cancer (MCF-7) cells in vitro at concentrations as low as 10 nM, with a concentration of 100 nM achieving proliferative effects similar to those of 1 nM estradiol. Expression of the estrogen-responsive gene pS2 was also induced in MCF-7 cells in response to treatment with a concentration of genistein as low as 1 microM. At higher concentrations (above 20 microM), genistein inhibits MCF-7 cell growth. In vivo, we have shown that dietary treatment with genistein (750 ppm) for 5 days enhanced mammary gland growth in 28-day-old ovariectomized athymic mice, indicating that genistein acts as an estrogen in normal mammary tissue. To evaluate whether the estrogenic effects observed in vitro with MCF-7 cells could be reproduced in vivo, MCF-7 cells were implanted s.c. in ovariectomized athymic mice, and the growth of the estrogen-dependent tumors was measured weekly. Negative control animals received the American Institute of Nutrition (AIN)-93G diet, the positive control group received a new s.c. estradiol (2 mg) pellet plus the AIN-93G diet, and the third group received genistein at 750 ppm in the AIN-93G diet. Tumors were larger in the genistein (750 ppm)-treated group than they were in the negative control group, demonstrating that dietary genistein was able to enhance the growth of MCF-7 cell tumors in vivo. Increased uterine weights were also observed in the genistein-treated groups. In summary, genistein can act as an estrogen agonist in vivo and in vitro, resulting in the proliferation of cultured human breast cancer cells (MCF-7) and the induction of pS2 gene expression. Here we present new information that dietary genistein stimulates mammary gland growth and enhances the growth of MCF-7 cell tumors in ovariectomized athymic mice. PMID- 9731494 TI - Fractionated ionizing radiation accelerates loss of amplified MDR1 genes harbored by extrachromosomal DNA in tumor cells. AB - In tumor specimens such as those from neuroblastoma, ovarian, and lung carcinoma patients, the prevalence of extrachromosomal circular DNA molecules harboring amplified genes has been well established. In some cases, the amplified genes have been identified as oncogenes, and their increased expression appears to contribute to the maintenance and progression of the malignancy. The aim of this study was to investigate the effect of fractionated radiation treatment, given in daily doses similar to those administered clinically, on the stability of extrachromosomal circular DNA molecules in cancer cells. Our studies were conducted with multidrug-resistant KB cells, which harbor extrachromosomal copies of the multidrug resistance gene (MDR1) almost exclusively on circular DNA molecules of approximately 750 and 1500 kb pairs. This size range is representative of extrachromosomal circular DNA molecules that have been shown to harbor amplified oncogenes in vivo. Exponentially growing MDR KB cells were exposed to 1400 and 2800 cGy ionizing radiation administered in 7 and 14 fractions, respectively, at 200 cGy per fraction/day. A statistically significant decrease in MDR1 extrachromosomal gene copy number was reproducibly detected in the irradiated cells compared with unirradiated cells passaged for the duration of the experiment in the absence of radiation treatment. This decrease was accompanied by a reduction in multidrug resistance and in P-glycoprotein levels, as determined by clonogenic dose-response assays and Western analyses, respectively. P-glycoprotein is a multidrug transporter encoded by the MDR1 gene. Fluorescence in situ hybridization studies further determined that extrachromosomal circular DNA loss correlated to the entrapment of these DNA molecules in radiation-induced micronuclei. These results indicate that radiation induced loss of extrachromosomally amplified genes from tumor cells via their entrapment in micronuclei contributes to the improved therapeutic response observed for some cancers. PMID- 9731495 TI - Irreversible G2-M arrest and cytoskeletal reorganization induced by cytotoxic nucleoside analogues. AB - The mechanisms by which cytotoxic agents perturb the normal cell biology and cell cycle progression of cancer cells were explored using B16F10 cells genetically modified to express the Herpes Simplex Virus-thymidine kinase gene. Culture in the presence of the nucleoside analogue ganciclovir induced a profound morphological change that required entry of treated cells into S phase and was dependent on prenylated proteins such as those of the rho gene family. Cell cycle arrest occurred in late S phase or G2 phase due to the activation of the G2-M DNA damage checkpoint. This checkpoint control operated at the level of inhibition of the activity of Cdc2/cyclin B and occurred by two mechanisms: (a) p53-mediated up regulation of p21CIP/WAF1 expression and its association with Cdc2/cyclin B; and (b) prevention of the dephosphorylation of tyrosine 15 of Cdc2. These events occurred in vitro and in vivo, and were shown to mediate bystander killing in this model. The mechanism of cell death seemed to be due to the irreversible cell cycle arrest at the G2-M checkpoint, rather than induction of apoptosis. These data link DNA damage checkpoints with cytoskeletal signaling pathways and the core cell cycle machinery and may represent a general mechanism of cytotoxicity of this class of nucleoside analogues. PMID- 9731496 TI - Tumor targeting with biotinylated tumor necrosis factor alpha: structure-activity relationships and mechanism of action on avidin pretargeted tumor cells. AB - We have recently described a new strategy for targeting biotinylated tumor necrosis factor-alpha (TNF-alpha) to tumors, based on pretargeting with biotinylated antibodies and avidin. Here, we have analyzed the structure-activity relationships of several biotin-TNF-alpha conjugates and studied the mechanism of their interaction with avidin and TNF-alpha receptors on tumor cells. The study has been carried out using an in vitro model based on human melanoma Colo 38 cells and monoclonal antibody 225, an antibody against the high molecular weight melanoma-associated antigen. Immunochemical and cytotoxicity studies showed that biotin-TNF-alpha but not TNF-alpha persists for several hours on the surface of cells pretargeted with biotin-monoclonal antibody 225 and avidin and triggers cytolytic effects. Studies on the mechanism of action showed that biotin-TNF alpha trimers can slowly dissociate from targeted cells in a bioactive form, through trimer-monomer-trimer transitions. Structure-activity relationship studies showed that nonbiotinylated subunits must be present in the biotin-TNF alpha trimers for efficient release of bioactive TNF-alpha. Colo 38 cells targeted with biotin-TNF-alpha were able to kill mouse L-M cells in coculture experiments, indicating that the released TNF-alpha can interact also with TNF alpha receptors expressed by bystander cells. In conclusion, the targeting complex works as a system that slowly releases bioactive TNF-alpha in the microenvironment of the targeted cell. This opens up the possibility that cells other than those reached by the targeting antibody (e.g., endothelial cells and local cells of the immune system) can be affected in vivo. PMID- 9731497 TI - Superior cytotoxicity with ganciclovir compared with acyclovir and 1-beta-D arabinofuranosylthymine in herpes simplex virus-thymidine kinase-expressing cells: a novel paradigm for cell killing. AB - Enzyme-prodrug therapy using ganciclovir and herpes simplex virus-thymidine kinase (HSV-TK) has demonstrated excellent antitumor activity in many different types of malignant cells. Previously, we noted that ganciclovir was substantially more cytotoxic than other HSV-TK substrates. Therefore, we embarked on a study to determine the basis for the superior cytotoxicity of ganciclovir. In U251tk human glioblastoma cells that stably express HSV-TK, ganciclovir elicited a >4 log cell kill instead of the < or =1.5 log cell kill mediated by two other HSV-TK substrates, 1-beta-D-arabinofuranosylthymine (araT) and acyclovir. Study of the metabolism of these drugs demonstrated that acyclovir was poorly phosphorylated to its active triphosphate with DNA incorporation below the limit of detection, which may explain the < 1 log cell kill in these cells. Lower levels of ganciclovir triphosphate accumulated compared with araT triphosphate (araTTP) under conditions that induced < or =1 log cell kill (67 versus 1235 pmol/10(7) cells, respectively), and the half-life for the triphosphate of ganciclovir was shorter than that of araT (terminal half-lives of 15 and 41 h, respectively). Incorporation of ganciclovir monophosphate into DNA was less than that of araT monophosphate, and both analogues were retained in DNA for > or =48 h. Thus, the superior cytotoxicity of ganciclovir was not due to enhanced metabolism to active forms. Highly cytotoxic concentrations of ganciclovir produced only weak inhibition of DNA synthesis. This allowed cells to proceed through S and G2-M phases during and after drug exposure, resulting in a doubling of cell number by 48 h after drug washout. As they attempted to progress through the cell cycle a second time, ganciclovir-treated cells accumulated in early S-phase and remained there until cell death, suggesting that ganciclovir incorporation in the DNA template was important for cytotoxicity. In contrast, strong inhibition of DNA synthesis by araTTP prevented cells from traversing the cell cycle for at least 12 h after drug washout, when the active metabolite was largely degraded araT treated cells were unable to divide for at least 72 h after drug exposure, at which point the surviving cells displayed a normal cell cycle distribution pattern. Based on the results presented here, we propose a novel paradigm in which the ability of ganciclovir to incorporate into DNA without inhibiting progression through S-phase, combined with high cytotoxicity for incorporated ganciclovir monophosphate, produces multilog cytotoxicity. PMID- 9731498 TI - Lysosomal sequestration of polyamine analogues in Chinese hamster ovary cells resistant to the S-adenosylmethionine decarboxylase inhibitor, CGP-48664. AB - CGP-48664, an inhibitor of the polyamine biosynthetic enzyme S-adenosylmethionine decarboxylase (AdoMetDC), is presently undergoing Phase 1 clinical trials as an experimental anticancer agent. We have shown previously (D. L. Kramer et al., J. Biol. Chem., 270: 2124-2132, 1995) that Chinese hamster ovary (CHO) cells that are made resistant to the growth inhibitory effects of the drug overexpress AdoMetDC because of a stable gene amplification. Unexpectedly, these same cells (CHO/644) were found to be insensitive to the growth inhibitory effects of N1,N11 diethylnorspermine (DENSPM)-a polyamine analogue also undergoing Phase 1 clinical trials-despite accumulating approximately 5 times more analogue than parental cells. We now report that treatment of CHO/664 cells with DENSPM results in the formation of numerous large cytoplasmic vacuoles, which on the basis of electron microscopy and cytochemical staining seem to be lysosomal in origin. A series of newly established CHO cell lines made differentially resistant to 1, 3, 10, 30, and 100 microM CGP-48664 by chronic exposure were used to demonstrate that vacuole formation correlated with the accumulation of extremely high levels of DENSPM without increasing growth inhibition. These same cells were used to show that AdoMetDC gene overexpression as indicated by mRNA levels was unrelated to vacuole formation; cells resistant to 100 microM CGP-48664 displayed a 170-fold increase in AdoMetDC mRNA levels and formed vacuoles in response to DENSPM, whereas those resistant to 10 microM CGP-48664 displayed a 120-fold increase in AdoMetDC mRNA levels and failed to form vacuoles. Despite accumulating to high intracellular levels, DENSPM was much less effective than spermine at down regulating ornithine decarboxylase and polyamine transport activities in highly resistant cells. Similarly, DENSPM was less able to induce spermidine/spermine N1 acetyltransferase activity in cells that formed vacuoles than in those that did not. Overall, natural polyamines failed to induce vacuoles and various analogues of DENSPM were used to probe the structural specificity of the effect. The data are consistent with the probability that DENSPM is sequestered to high concentrations in lysosomal vacuoles of CGP-48664-resistant cells and is, therefore, not available to interact with polyamine regulatory sites or to cytotoxically affect cell growth. In addition to implicating the lysosome as a potential new site of CGP-48664 drug action that could be involved in antitumor activity and/or host toxicities, the findings also suggest a potential mechanism of cell resistance to analogues such as DENSPM. PMID- 9731499 TI - Allogeneic cell therapy for a murine mammary carcinoma. AB - The effect of allogeneic cell therapy on tumor growth was studied in a murine model of mammary carcinoma (4T1) as an experimental model of solid tumors in humans. i.v. inoculation of 4T1 (H-2d) cells into syngeneic mice [BALB/c or (BALB/cXC57BL/6)F1] (F1) carrying the H-2d histocompatible antigens results in tumor colonies in the lungs that finally cause the death of all of the mice. Sublethally irradiated F1 mice were inoculated with 4T1 cells to simulate minimal residual disease and with immunocompetent splenocytes derived from naive donors of F1 (syngeneic), BALB/c (syngeneic to the tumor but semiallogeneic to the host), or C57BL/6 (allogeneic to the tumor and semiallogeneic to the host) mice. The survival of F1 tumor-bearing mice that were treated with allogeneic C57BL/6 splenocytes was significantly prolonged (P < 0.02) compared with hosts given F1 or BALB/c-derived splenocytes that are syngeneic to 4T1 tumor cells. Adoptive transfer of lung cells that were isolated from F1 primary mice inoculated with 4T1 cells and syngeneic BALB/c or F1 splenocytes led to local tumor growth and death in secondary recipients. In contrast, only 1 of 22 secondary recipients developed tumors when inoculated with lung cells derived from F1 mice given allogeneic C57BL/6 splenocytes. All of the 21 secondary hosts survived disease free for a follow-up time of >200 days. These results indicate that immunocompetent cells allogeneic to the mammary carcinoma cells were able to inhibit tumor development in the primary hosts and to prevent tumor growth in the adoptive recipients, which suggests that allogeneic cell therapy may be an efficient antitumor tool to eradicate minimal residual disease in human solid tumors. PMID- 9731500 TI - Docetaxel chronopharmacology in mice. AB - Docetaxel tolerance and antitumor efficacy could be enhanced if drug administration was adapted to circadian rhythms. This hypothesis was investigated in seven experiments involving a total of 626 male B6D2F1 mice, synchronized with an alternation of 12 h of light and 12 h of darkness (12:12), after i.v. administration of docetaxel. In experiment (Exp) 1, the drug was given once a week (wk) for 6 wks (20 mg/kg/wk) or for 5 wks (30 mg/kg/wk) at one of six circadian times, during light when mice were resting [3, 7, or 11 hours after light onset (HALO)], or during darkness, when mice were active (15, 19, or 23 HALO). Endpoints were survival and body weight change. In Exp 2 and 3, docetaxel (30 mg/kg/wk) was administered twice, 1 wk apart, at one of four circadian stages (7, 11, 19, or 23 HALO). Endpoints were hematological and intestinal toxicities. In Exp 4, circadian changes in cell cycle phase distribution and BCL-2 immunofluorescence were investigated in bone marrow as possible mechanisms of docetaxel tolerability rhythm. In Exp 5 to 7, docetaxel was administered to mice bearing measurable P03 pancreatic adenocarcinoma (270-370 mg), with tumor weight and survival as endpoints. Mice from Exp 5 and 6 received a weekly schedule of docetaxel at one of six circadian stages (20 or 30 mg/kg/wk at 3, 7, 11, 15, 19, or 23 HALO). In Exp 7, docetaxel (30 mg/kg) was given every 2 days (day 1, 3, 5 schedule) at 7, 11, 19, or 23 HALO. Docetaxel dosing in the second half of darkness (19 or 23 HALO) resulted in significantly worse toxicity than its administration during the light span (3, 7, or 11 HALO). The survival rate ranged from 56.3% in the mice treated at 23 HALO to 93.8 or 87.5% in those injected at 3 or 11 HALO, respectively (Exp 1, P < 0.01). Granulocytopenia at nadir was -49 +/- 14% at 7 HALO compared with -84 +/- 3% at 19 HALO (Exp 2 and 3, P < 0.029), and severe jejunal mucosa necrosis occurred in 5 of 8 mice treated at 23 HALO as opposed to 2 of 18 receiving docetaxel at 7, 11, or 19 HALO (Exp 2 and 3, P < 0.02). The time of least docetaxel toxicity corresponded to the circadian nadir in S or G2-M phase and to the circadian maximum in BCL-2 immunofluorescence in bone marrow. Docetaxel increased the median survival of tumor-bearing mice in a dose-dependent manner (controls: 24 days; 20 mg/kg weekly, 33 days; 30 mg/kg weekly or day 1, 3, 5 schedule, 44 or 46 days, respectively; Exp 5-7). Survival curves of treated mice differed significantly according to dosing time for each dose and schedule (P from log rank <0.003 to P < 0.03). In Exp 5 and 6, the percentage of increase in life span was largest if docetaxel was administered weekly at 7 HALO (20 mg/kg, 220%; 30 mg/kg, 372%) and lowest after docetaxel dosing at 19 HALO (80% with 20 mg/kg) or at 15 HALO (78% with 30 mg/kg). In Exp 7, (day 1, 3, 5 schedule), docetaxel was most active at 11 HALO (percentage increase in life span, 390%) and least active at 23 HALO (210%). Docetaxel tolerability and antitumor efficacy were simultaneously enhanced by drug dosing in the light span, when mice were resting. Mechanisms underlying the tolerability rhythm likely involved the circadian organization of cell cycle regulation. Docetaxel therapeutic index may be improved with an administration at night in cancer patients, when fewest bone marrow cells are in S or G2-M phase. PMID- 9731501 TI - Chimeric human-mouse IgG antibodies with shuffled constant region exons demonstrate that multiple domains contribute to in vivo half-life. AB - Structural features that determine the differing rates of immunoglobulin catabolism are of great relevance to the engineering of immunologically active reagents. Sequences in the CH2 and CH3 region of IgG have been shown to regulate the rate of clearance through their interaction with FcRn. In an attempt to probe additional structural features that regulate antibody half-life, we have investigated two families of chimeric antibodies, composed of identical murine heavy and light antidansyl variable regions joined to human kappa light-chains and wild-type or shuffled human IgG heavy-chain constant regions. These antibodies were iodinated, and their clearance was studied in severe combined immunodeficient mice hosts by whole-body radioactivity measurements. Clearances of the wild-type and recombinant antibodies were biphasic. In a panel of immunoglobulins derived from IgG2 and IgG3, as successive domains were varied from gamma2 to gamma3, beta-phase half-life gradually decreased from 337.0 h to 70.6 h. Statistical analysis suggested that the composition of each of the three domains affected half-life, and no single region of the molecule by itself determined the rate of clearance. In the second panel of immunoglobulins derived from IgG1 and IgG4, the construct with the amino terminus portion of the molecule derived from IgG4, joined within the CH2 domain to the COOH terminus portion of IgG1, had a half-life paradoxically greater than either IgG1, or IgG4 (P < 0.012). All four IgG1/IgG4 constructs demonstrated presence of the concentration catabolism phenomenon, which is a unique hallmark of immunoglobulin catabolism. The contribution of all three constant region domains to immunoglobulin half-life may be due to distant conformational effects in addition to direct binding to protective receptors, and emphasizes the importance of distant sequences on the rate of immunoglobulin catabolism. Interesting possibilities regarding mechanisms controlling immunoglobulin metabolism are raised by the hybrid gamma4/gamma1 molecule with a half-life greater than either parental immunoglobulin. Understanding the relationships between the structure of these molecules and their clearance rate will further our ability to produce immunoglobulins with improved pharmacokinetic properties. PMID- 9731502 TI - Antigen expressed on tumor cells fails to elicit an immune response, even in the presence of increased numbers of tumor-specific cytotoxic T lymphocyte precursors. AB - We have used T-cell receptor (TCR) transgenic mice to analyze the interaction of tumors with the immune system. We show that the tumor cell line Lewis lung lymphocytic choriomeningitis virus (LL-LCMV), genetically manipulated to express an H-2 Db-restricted epitope of the lymphocytic choriomeningitis virus glycoprotein (LCMV33-41), can grow progressively in TCR transgenic mice, where approximately 50% of CD8+ T cells are specific for LCMV33-41. TCR transgenic T cells were not deleted in tumor-bearing mice, and their surface phenotype and cytokine secretion patterns remained typical of naive T cells. Also, TCR transgenic T cells from tumor-bearing mice had undiminished capacity to proliferate to antigen in vitro. Tumor-protective immune responses could be elicited in TCR transgenic mice by immunization with LCMV33-41 peptide-loaded dendritic cells. Tumor resistance correlated with the switch of TCR transgenic T cells from a CD44low to a CD44high phenotype and increased capacity to produce IFNgamma in vitro. Results similar to those obtained in TCR transgenic mice were also obtained using an adoptive transfer system, where small numbers of TCR transgenic T cells were injected into normal C57BL/6 hosts. These data indicate that even large tumors may not induce specific immunization, tolerance, or anergy to tumor antigens, and that high numbers of tumor-specific CTL precursors are not sufficient to provide tumor resistance. PMID- 9731503 TI - Induction of persistent tumor-protective immunity in mice cured of established colon carcinoma metastases. AB - The induction of tumor-specific T-cell responses that are effective in eradicating disseminated tumors and in mounting a persistent tumor-protective immunity is one of the major goals of tumor immunotherapy. Here, we demonstrate that we achieved this goal by directing interleukin 2 (IL-2) to the tumor microenvironment of colon carcinoma metastases in syngeneic mice with a recombinant antibody-IL-2 fusion protein (huKS1/4-IL-2). Eradication of established pulmonary metastases is induced by a CD8+ T cell-mediated immune response, which can be transmitted to naive syngeneic severe combined immunodeficient mice by adoptive transfer of CD8+ T cells from immune animals. This immune response was followed by the induction of a long-lived immunity against challenge up to 5 months later with CT26-KSA or wild-type CT26 murine colon carcinoma cells in BALB/c mice. This memory immune response was confirmed by flow cytometric analyses of CD8+ T cells isolated from secondary lymphoid tissue that revealed a phenotypic profile typical of early memory T cells. This long-lived protective tumor immunity was successfully boosted to become optimally effective in all experimental animals by injections of noncurative doses of IL-2 fusion protein 4 days after challenge with tumor cells. Taken together, our results indicate that the huKS1/4-IL-2 fusion protein elicits a long-lived cellular memory immune response that can be amplified by additional applications of IL-2 fusion proteins. This approach could become useful for the treatment of colorectal carcinoma in an adjuvant setting, particularly in patients with minimal residual disease. PMID- 9731504 TI - The human p19ARF protein encoded by the beta transcript of the p16INK4a gene is frequently lost in small cell lung cancer. AB - The p16IN4/CDKN2/MTS1 gene encodes two structurally different proteins: a cyclin dependent kinase inhibitor called p16INK4a, which regulates retinoblastoma protein-dependent G1 arrest, and a cell cycle inhibitor designated p19ARF, which arrests cell growth in G1-S and also in G2-M. Whereas inactivation of p16INK4a has been described as a frequent event in lung cancer, the current function of p19ARF is still poorly understood. We have examined the expression of the human p19ARF (hp19ARF) protein in a large series of lung cancers using immunohistochemistry and showed that the protein was more frequently lost in high grade neuroendocrine (NE) lung tumors (large cell NE carcinoma and small cell lung carcinoma; 51 of 78, 65%) than it was in non-small cell lung cancer (25 of 101, 25%). No deleterious mutation was found in exons 1beta and 2 of hp19ARF in those NE tumors with negative immunoreactivity, and a beta transcript was detected in the majority of them. Concomitant absence of hp19ARF and retinoblastoma proteins was frequently detected in high-grade NE lung tumors, whereas no relationship could be found between the status of hp19ARF and p53 proteins in those tumors. These results are consistent with an alternative growth suppressor function for hp19ARF in NE lung cancer that is distinct from that of p16INK4a. Moreover, the frequent uncoupling between the beta transcript and the hp19ARF protein suggests a novel mechanism of inactivation at the translational level. PMID- 9731505 TI - Expression of the ErbB-neuregulin signaling network during human cerebellar development: implications for the biology of medulloblastoma. AB - The four receptor tyrosine kinase I receptors, ErbB-1, ErbB-2, ErbB-3, and ErbB 4, which have been implicated in the development of a variety of normal and malignant tissues, are activated through ligand mediated homo- and heterodimerization. We have previously reported the frequent coexpression, heterodimerzation, and prognostic significance of ErbB-2 and ErbB-4 in childhood medulloblastoma, an embryonal tumor of the cerebellar external granule cell layer (EGL). In the present study, we have used immunohistochemistry and Western blotting analysis to analyze the expression of the ErbB receptors and neuregulin (NRG) 1-alpha and NRG1-beta ligands during normal human cerebellar development. We demonstrate that ErbB-1, ErbB-3, ErbB-4, and NRG1-beta display specific temporal and topographical distribution in the cerebellum during intrauterine and postnatal life, and that normal ErbB-NRG signaling in the EGL multiplying zone is likely to be mediated by ErbB-4 and NRG1-beta. In contrast, ErbB-2, which is expressed in 86% of medulloblastomas, could not be detected at any stage of cerebellar development. Therefore, we propose that positive deregulation of ErbB 2 expression in the cerebellar EGL, leading to the formation of a NRG41-beta driven ErbB-2/ErbB-4 autocrine loop, is an important factor in medulloblastoma tumorigenesis. In further support of this hypothesis, we provide evidence using reverse transcription-PCR analysis that expression of the ErbB-2 and ErbB-4 receptors, but not ErbB-1 or ErbB-3, is deregulated in medulloblastoma compared with normal developing cerebellum. We also demonstrate NRG1-beta expression in 87% (n = 46 of 48) of medulloblastoma primary tumors, with the greatest expression levels occurring in tumors with high ErbB-2 and ErbB-4 receptor coexpression. Furthermore, the expression of all three components of the proposed autocrine loop (ie., ErbB-2, ErbB-4, and NRG1-beta) was significantly related to the presence of metastases at diagnosis (P < 0.05). PMID- 9731506 TI - Hypermethylation of the p16INK4a promoter in colectomy specimens of patients with long-standing and extensive ulcerative colitis. AB - Functional inactivation of the p16INK4a gene has been reported to be involved in the development of a variety of human malignancies. Recent evidence shows that transcriptional silencing as a consequence of hypermethylation of CpG islands is the predominant mechanism of p16INK4a gene inactivation in sporadic colon cancer. This study sought to identify the significance of p16INK4a methylation in the colonic epithelium of patients with long-standing ulcerative colitis. A total of 89 tissue samples was retrieved from three colectomy specimens. A methylation specific PCR assay was applied. The methylation status was compared with histological findings and the flow cytometrically determined DNA index. Hypermethylation of the p16INK4a promoter region was detected in 12.7% of samples that were negative for dysplasia. However, 70.0% of samples with dysplasia and all of the samples with carcinomatous lesions revealed hypermethylation. Hypermethylation of the p16INK4a gene promoter was detected already in 40% of specimens with lesions indefinite for dysplasia and in 13.7% of samples with exclusively diploid cell populations. These results suggest that hypermethylation of the p16INK4a promoter region is a frequent and early occurring event during the process of neoplastic progression in ulcerative colitis. PMID- 9731507 TI - Mutation rate of a microsatellite sequence in normal human fibroblasts. AB - Dinucleotide repeats, because of their repetitive nature, are prone to frameshift mutations, most likely via a DNA-polymerase slippage mechanism. Mutation rates in microsatellite DNA sequences are high in mismatch repair-defective cells. In normal cells, only estimates of maximal rates of mutation in microsatellites have been possible previously, because of the low sensitivity of screening assays for mutations in endogenous sequences. We have measured the spontaneous mutation rate of a dinucleotide repeat in diploid human foreskin fibroblasts. In our system, the mutation target is a (CA)17 repeat contained within a stably integrated plasmid. The repeat disrupts the reading frame of a neomycin (neo) resistance gene within the plasmid. Cells containing frameshift mutations in the CA repeat that correct the reading frame of the neo gene are selected using the neo analogue G418. This system of measuring microsatellite mutation rates is highly sensitive, because there is a specific target within which mutations can be selected. Fluctuation analysis of cells containing the target DNA yielded mutation rates of <3.1 x 10(-8) to 44.8 x 10(-8) mutations/cell/generation. This is the first report of a direct measurement of a spontaneous mutation rate of a microsatellite sequence in normal human cells. PMID- 9731508 TI - Regulation of expression of the DNA repair gene O6-methylguanine-DNA methyltransferase via protein kinase C-mediated signaling. AB - O6-Alkylguanine is the major mutagenic and cytotoxic DNA lesion induced by alkylating agents, including 2-chloroethyl-N-nitrosourea-based antitumor drugs. This lesion is repaired by O6-methylguanine-DNA methyltransferase (MGMT), the expression of which is highly variable in both normal tissues and in tumor cells. The promoter of the human MGMT gene was found to contain two putative activator protein (AP)-1 sites. Here, we show that the level of MGMT mRNA in HeLa S3 cells was increased 3-5-fold by phorbol-12-myristate-13-acetate (TPA) and 1,2-diacyl-sn glycerol (DAG), which are activators of protein kinase C (PKC), as well as by okadaic acid, an inhibitor of protein phosphatases. The PKC inhibitor 1-(5 isoquinoline sulfonyl)-2-methylpiperazine-HCl eliminated MGMT activation by TPA and DAG but not by OA. Prior down-regulation of PKC abolished subsequent effects of TPA or DAG. The results indicate AP-1 to be involved in regulation of MGMT expression. This hypothesis was supported by showing AP-1 binding to two target sequences of the MGMT promoter and transactivation of the MGMT promoter upon cotransfection with c-fos and c-jun in F9 cells. That TPA-mediated induction of MGMT caused increased cellular resistance to 2-chloroethyl-N-nitrosourea suggests a therapeutic significance for PKC-mediated MGMT modulation. PMID- 9731509 TI - Improved quantitation of minimal residual disease in multiple myeloma using real time polymerase chain reaction and plasmid-DNA complementarity determining region III standards. AB - The complementarity determining region III of the rearranged immunoglobulin heavy chain gene has been the target for tumor-specific PCR assays for the detection and follow-up of B-cell malignancies. Previously, these assays have relied on gel based end point data collection methods (i.e., band densitometry) and, thus, have provided at best a semiquantitative assessment of tumor levels. We show the development of a novel, real-time TaqMan PCR assay to quantitate residual multiple myeloma cells in clinical samples after high-dose chemotherapy and autologous stem cell transplantation. We provide evidence that real-time PCR is reproducible, sensitive, and quantitative. In a 40-replicate PCR experiment targeting the beta-actin gene, the coefficient of variation for threshold cycle data was 1.6%, whereas it increased to 13.6% and 31%, respectively, for end point fluorescence and gel densitometry. Moreover, in an experiment directly comparing standard curves obtained from band densitometry and threshold cycle data, the standard curve constructed from threshold cycle data had a multiple R2 value of 1.00 and demonstrated a dynamic range >4 logs, compared with the 2-log linear range of gel densitometry. Finally, we show that when a complementarity determining region III-specific PCR primer is used in conjunction with a consensus primer for the immunoglobulin heavy chain joining gene, plasmid DNA can be used as a readily available and effective substitute for clonal plasma-cell genomic DNA when preparing standards. By applying real-time PCR to the analysis of clinical samples, we are able to quantitate levels of tumor involvement with unparalleled reproducibility and statistical confidence. Real-time PCR technology may well provide the accuracy and reliability necessary for minimal residual disease detection to have real prognostic significance. PMID- 9731510 TI - Mammalian 3-methyladenine DNA glycosylase protects against the toxicity and clastogenicity of certain chemotherapeutic DNA cross-linking agents. AB - DNA repair status is recognized as an important determinant of the clinical efficacy of cancer chemotherapy. To assess the role that a mammalian DNA glycosylase plays in modulating the toxicity and clastogenicity of the chemotherapeutic DNA cross-linking alkylating agents, we compared the sensitivity of wild-type murine cells to that of isogenic cells bearing homozygous null mutations in the 3-methyladenine DNA glycosylase gene (Aag). We show that Aag protects against the toxic and clastogenic effects of 1,3-bis(2-chloroethyl)-1 nitrosourea and mitomycin C (MMC), as measured by cell killing, sister chromatid exchange, and chromosome aberrations. This protection is accompanied by suppression of apoptosis and a slightly reduced p53 response. Our results identify 3-methyladenine DNA glycosylase-initiated base excision repair as a potentially important determinant of the clinical efficacy and, possibly, the carcinogenicity of these widely used chemotherapeutic agents. However, Aag does not contribute significantly to protection against the toxic and clastogenic effects of several chemotherapeutic nitrogen mustards (namely, mechlorethamine, melphalan, and chlorambucil), at least in the mouse embryonic stem cells used here. We also compare the Aag null phenotype with the Fanconi anemia phenotype, a human disorder characterized by cellular hypersensitivity to DNA cross-linking agents, including MMC. Although Aag null cells are sensitive to MMC-induced growth delay and cell cycle arrest, their sensitivity is modest compared to that of Fanconi anemia cells. PMID- 9731511 TI - Centrosome defects and genetic instability in malignant tumors. AB - Genetic instability is a common feature of many human cancers. This condition is frequently characterized by an abnormal number of chromosomes, although little is known about the mechanism that generates this altered genetic state. One possibility is that chromosomes are missegregated during mitosis due to the assembly of dysfunctional mitotic spindles. Because centrosomes are involved in spindle assembly, they could contribute to chromosome missegregation through the organization of aberrant spindles. As an initial test of this idea, we examined malignant tumors for centrosome abnormalities using antibodies to the centrosome protein pericentrin. We found that centrosomes in nearly all tumors and tumor derived cell lines were atypical in shape, size, and composition and were often present in multiple copies. In addition, virtually all pericentrin-staining structures in tumor cells nucleated microtubules, and they participated in formation of disorganized mitotic spindles, upon which chromosomes were missegregated. All tumor cell lines had both centrosome defects and abnormal chromosome numbers, whereas neither was observed in nontumor cells. These results indicate that centrosome defects are a common feature of malignant tumors and suggest that they may contribute to genetic instability in cancer. PMID- 9731512 TI - Genomic instability and catalase gene amplification induced by chronic exposure to oxidative stress. AB - Chronic exposure (>200 days) of HA1 fibroblasts to increasing concentrations of H2O2 or O2 results in the development of a stable oxidative stress-resistant phenotype characterized by increased cellular antioxidant levels, particularly catalase (D. R. Spitz et al, Arch. Biochem. Biophys., 279: 249-260, 1990; D. R. Spitz et al., Arch. Biochem. Biophys., 292: 221-227, 1992; S. J. Sullivan et al., Am. J. Physiol. (Lung Cell. Mol. Physiol.), 262: L748-L756, 1992). Acutely stressed cells failed to develop a stably resistant phenotype or increased catalase activity, suggesting that chronic exposure is required for the development of this phenotype. This study investigates the mechanism underlying increased catalase activity in the H2O2- and O2-resistant cell lines. In H2O2- and O2-resistant cells, catalase activity was found to be 20-30-fold higher than that in the parental HA1 cells and correlated with increased immunoreactive catalase protein and steady-state catalase mRNA levels. Resistant cell lines also demonstrated a 4-6-fold increase in catalase gene copy number by Southern blot analysis, which is indicative of gene amplification. Chromosome banding and in situ hybridization studies identified a single amplified catalase gene site located on a rearranged chromosome with banding similarities to Z-4 in the hamster fibroblast karyotype. Simultaneous in situ hybridization with a Z-4 specific adenine phosphoribosyltransferase (APRT) gene revealed that the amplified catalase genes were located proximate to APRT on the same chromosome in all resistant cells. In contrast, HA1 cells contained only single copies of the catalase gene that were not located on APRT-containing chromosomes, indicating that amplification is associated with a chromosomal rearrangement possibly involving Z-4. The fact that chronic exposure of HA1 cells to either HO2 or 95% O2 resulted in gene amplification suggests that gene amplification represents a generalized response to oxidative stress, contributing to the development of resistant phenotypes. These results support the hypothesis that chronic exposure to endogenous metabolic or exogenous environmental oxidative stress represents an important factor contributing to gene amplification and genomic instability. PMID- 9731513 TI - Human T-cell leukemia virus type I Tax transactivates human interleukin 8 gene through acting concurrently on AP-1 and nuclear factor-kappaB-like sites. AB - The transactivator protein, Tax, from the human T-cell leukemia virus type I (HTLV-I) transactivates both viral and cellular genes. Previously, we had shown that interleukin 8 (IL-8) is constitutively expressed in HTLV-I-infected cells and in cells transiently expressing Tax. We show here that the IL-8 promoter is Tax responsive in Jurkat T cells. Furthermore, using several deletion and mutated plasmids of the 5'-flanking regulatory region of the IL-8 gene linked to the luciferase gene as a reporter and mutant tax gene expression vectors, we have established that both AP-1 at -126 to -120 and nuclear factor (NF)-kappaB-like cis-element at -80 to -71 are essential and sufficient for the induction of the IL-8 gene by HTLV-I Tax. In addition, overexpression of the dominant-negative mutants of NF-kappaB inhibitor molecules, IkappaBalpha and IkappaBbeta, abolished the Tax-induced activation of IL-8 gene. Gel mobility shift assays detected proteins specifically binding to the AP-1 and NF-kappaB-like sites in Tax expressing T-cell lines infected with HTLV-I. Similarly, the nuclear translocation of proteins specifically bound to these two motifs was shown in JPX 9 cells, a subclone of Jurkat cells, carrying the Tax sequences under the control of an inducible promoter. Taken together, these results suggest that the cooperation of transcription factors NF-kappaB and AP-1 is essential for transactivation of IL-8 gene by HTLV-I Tax. PMID- 9731514 TI - Synthetic matrix metalloproteinase inhibitors and tissue inhibitor of metalloproteinase (TIMP)-2, but not TIMP-1, inhibit shedding of tumor necrosis factor-alpha receptors in a human colon adenocarcinoma (Colo 205) cell line. AB - The solubilization of plasma membrane receptors through proteolytic cleavage of the ligand binding domain at the cell surface is an important mechanism for regulating cytokine function and receptor signaling. The inhibition of the shedding of a variety of receptors by synthetic inhibitors of the matrix metalloproteinases (MMPs) implicates metalloproteinases in this regulatory event. We examined the effects of two naturally occurring tissue inhibitors of metalloproteinases, TIMP-1 and TIMP-2, and several synthetic MMP inhibitors (MMPIs) on the shedding of both tumor necrosis factor alpha receptor type I (TNFalpha-RI; Mr 55,000) and TNFalpha-RII (Mr 75,000) by the Colo 205 human colon adenocarcinoma cell line. Culture of Colo 205 cells for 48 h resulted in the shedding of both TNFalpha-RI and TNFalpha-RII, as determined by ELISA. The shedding of TNFalpha receptors was not affected by TIMP-1 or protease inhibitors aprotinin, pepstatin, or leupeptin but was inhibited in a dose-dependent manner by the following synthetic MMPIs: batimastat and marimastat (BB-94 and BB-2516, respectively, British Biotech, Inc.); CT1418 (Celltech Therapeutics); CGS27023A (Novartis Pharmaceuticals); and RO31-9790 (Roche), with IC50s ranging from 3.2 to 38.0 microM. Similarly, TIMP-2 from two different sources reproducibly inhibited the shedding of both TNFalpha-RI and TNFalpha-RII in a dose-dependent manner (IC50 = 286 +/- 33 nM for TNFalpha-RI shedding and 462 +/- 52 nM for shedding of TNFalpha-RII). The inhibition of TNFalpha-RI shedding was confirmed in the SW626 human ovarian adenocarcinoma cell line. The synthetic MMPIs and TIMP-2, but not TIMP-1, also caused a dose-dependent increase in the number of TNFalpha receptors retained on the surface of Colo 205 cells, as determined by flow cytometry. Inhibition of TNFalpha receptor shedding with TIMP-2 occurs at molar concentrations 10-100 times less than those required with low molecular weight, synthetic MMPIs but at concentrations greater than those required to inhibit collagen degradation. Modulation of TNFalpha receptor shedding by TIMP-2 could have important implications for the pleiotropic effects of TNFalpha in both normal and malignant cells and for the pharmacological activity of synthetic MMPIs. PMID- 9731515 TI - Regulation of vascular endothelial growth factor expression in human colon cancer by insulin-like growth factor-I. AB - We investigated the role of insulin-like growth factor (IGF)-I and IGF-binding proteins (IGFBPs) in the regulation of vascular endothelial growth factor (VEGF) expression in colon cancer cells and the mechanism by which this regulation occurs. HT29 human colon cancer cells were treated with IGF-I for various time periods. VEGF mRNA expression increased within 2 h and peaked at 24 h. SW620 colon cancer cells exhibited a peak induction of VEGF mRNA 8 h after IGF-I treatment. IGF-I induction of VEGF was confirmed at the protein level. In experiments using transient transfection of VEGF promoter-reporter constructs into HT29 cells, IGF-I increased the activity of the VEGF promoter, and pretreatment of HT29 cells with dactinomycin abrogated the induction of VEGF mRNA by IGF-I. The half-life of VEGF mRNA was not prolonged by treatment with IGF-I. Blocking the activity of IGFBP-4 did not significantly modulate the effect of IGF I induction of VEGF mRNA in HT29 cells. Treating cells with des-(1-3)-IGF-I (an active derivative of IGF-I that does not bind to binding proteins) had effects on VEGF mRNA expression that were similar to those of IGF-I. These findings suggest that IGF-I regulates VEGF expression in human colon cancer cells by induction of transcription of the VEGF gene. IGFBPs do not significantly affect IGF-I induction of VEGF. PMID- 9731516 TI - Correspondence re: Y.E. Shi et al., Antitumor activity of the novel human breast cancer growth inhibitor, mammary-derived growth inhibitor-related gene, MRG. Cancer Res., 57: 3084-3091, 1997. PMID- 9731517 TI - Conditionally yours. PMID- 9731518 TI - Massive attack on high-throughput biology. PMID- 9731519 TI - Rapid flux in plant genomes. PMID- 9731521 TI - The war of the sex chromosomes. PMID- 9731520 TI - A lungful of transcription factors. PMID- 9731522 TI - The APCI1307K allele and breast cancer risk. PMID- 9731523 TI - We're off to see the genome. AB - We discuss some societal and legal ramifications of the human genetics revolution. Our reflections were stimulated by discussions among scientists, citizens and legal experts at a large public symposium. We outline key issues regarding oversight of genetic research on human subjects, banking of DNA data by governments and corporations, the potential impact of behavioural genetics and effects upon racial and racist thinking. We contend that, in some cases, well intentioned but naive efforts to protect the rights of individuals and groups may hurt everyone by blocking the progress of useful research. PMID- 9731524 TI - Data management and analysis for gene expression arrays. AB - Microarray technology makes it possible to simultaneously study the expression of thousands of genes during a single experiment. We have developed an information system, ArrayDB, to manage and analyse large-scale expression data. The underlying relational database was designed to allow flexibility in the nature and structure of data input and also in the generation of standard or customized reports through a web-browser interface. ArrayDB provides varied options for data retrieval and analysis tools that should facilitate the interpretation of complex hybridization results. A sampling of ArrayDB storage, retrieval and analysis capabilities is available (www.nhgri.nih.gov/DIR/LCG/15K/HTML/ ), along with information on a set of approximately 15,000 genes used to fabricate several widely used microarrays. Information stored in ArrayDB is used to provide integrated gene expression reports by linking array target sequences with NCBI's Entrez retrieval system, UniGene and KEGG pathway views. The integration of external information resources is essential in interpreting intrinsic patterns and relationships in large-scale gene expression data. PMID- 9731525 TI - PAK3 mutation in nonsyndromic X-linked mental retardation. AB - Nonsyndromic X-linked mental retardation (MRX) syndromes are clinically homogeneous but genetically heterogeneous disorders, whose genetic bases are largely unknown. Affected individuals in a multiplex pedigree with MRX (MRX30), previously mapped to Xq22, show a point mutation in the PAK3 (p21-activated kinase) gene, which encodes a serine-threonine kinase. PAK proteins are crucial effectors linking Rho GTPases to cytoskeletal reorganization and to nuclear signalling. The mutation produces premature termination, disrupting kinase function. MRI analysis showed no gross defects in brain development. Immunofluorescence analysis showed that PAK3 protein is highly expressed in postmitotic neurons of the developing and postnatal cerebral cortex and hippocampus. Signal transduction through Rho GTPases and PAK3 may be critical for human cognitive function. PMID- 9731526 TI - Dysferlin, a novel skeletal muscle gene, is mutated in Miyoshi myopathy and limb girdle muscular dystrophy. AB - Miyoshi myopathy (MM) is an adult onset, recessive inherited distal muscular dystrophy that we have mapped to human chromosome 2p13. We recently constructed a 3-Mb P1-derived artificial chromosome (PAC) contig spanning the MM candidate region. This clarified the order of genetic markers across the MM locus, provided five new polymorphic markers within it and narrowed the locus to approximately 2 Mb. Five skeletal muscle expressed sequence tags (ESTs) map in this region. We report that one of these is located in a novel, full-length 6.9-kb muscle cDNA, and we designate the corresponding protein 'dysferlin'. We describe nine mutations in the dysferlin gene in nine families; five are predicted to prevent dysferlin expression. Identical mutations in the dysferlin gene can produce more than one myopathy phenotype (MM, limb girdle dystrophy, distal myopathy with anterior tibial onset). PMID- 9731527 TI - A gene related to Caenorhabditis elegans spermatogenesis factor fer-1 is mutated in limb-girdle muscular dystrophy type 2B. AB - The limb-girdle muscular dystrophies are a genetically heterogeneous group of inherited progressive muscle disorders that affect mainly the proximal musculature, with evidence for at least three autosomal dominant and eight autosomal recessive loci. The latter mostly involve mutations in genes encoding components of the dystrophin-associated complex; another form is caused by mutations in the gene for the muscle-specific protease calpain 3. Using a positional cloning approach, we have identified the gene for a form of limb girdle muscular dystrophy that we previously mapped to chromosome 2p13 (LGMD2B). This gene shows no homology to any known mammalian gene, but its predicted product is related to the C. elegans spermatogenesis factor fer-1. We have identified two homozygous frameshift mutations in this gene, resulting in muscular dystrophy of either proximal or distal onset in nine families. The proposed name 'dysferlin' combines the role of the gene in producing muscular dystrophy with its C. elegans homology. PMID- 9731528 TI - The paleontology of intergene retrotransposons of maize. AB - Retrotransposons, transposable elements related to animal retroviruses, are found in all eukaryotes investigated and make up the majority of many plant genomes. Their ubiquity points to their importance, especially in their contribution to the large-scale structure of complex genomes. The nature and frequency of retro element appearance, activation and amplification are poorly understood in all higher eukaryotes. Here we employ a novel approach to determine the insertion dates for 17 of 23 retrotransposons found near the maize adh1 gene, and two others from unlinked sites in the maize genome, by comparison of long terminal repeat (LTR) divergences with the sequence divergence between adh1 in maize and sorghum. All retrotransposons examined have inserted within the last six million years, most in the last three million years. The structure of the adh1 region appears to be standard relative to the other gene-containing regions of the maize genome, thus suggesting that retrotransposon insertions have increased the size of the maize genome from approximately 1200 Mb to 2400 Mb in the last three million years. Furthermore, the results indicate an increased mutation rate in retrotransposons compared with genes. PMID- 9731529 TI - Mass spectrometry and EST-database searching allows characterization of the multi protein spliceosome complex. AB - Many important cell mechanisms are carried out and regulated by multi-protein complexes, for example, transcription and RNA processing machinery, receptor complexes and cytoskeletal structures. Most of these complexes remain only partially characterized due to the difficulty of conventional protein analysis methods. The rapid expansion of DNA sequence databases now provides whole or partial gene sequences of model organisms, and recent advances in protein microcharacterization via mass spectrometry allow the possibility of linking these DNA sequences to the proteins in functional complexes. This approach has been demonstrated in organisms whose genomes have been sequenced, such as budding yeast. Here we report the first characterization of an entire mammalian multi protein complex using these methods. The machinery that removes introns from mRNA precursors--the spliceosome--is a large multi-protein complex. Approximately half of the components excised from a two-dimensional gel separation of the spliceosome were found in protein sequence databases. Using nanoelectrospray mass spectrometry, the remainder were identified and cloned using public expressed sequence tag (EST) databases. Existing EST databases are thus already sufficiently complete to allow rapid characterization of large mammalian protein complexes via mass spectrometry. PMID- 9731530 TI - Mutations in a polycistronic nuclear gene associated with molybdenum cofactor deficiency. AB - All molybdoenzymes other than nitrogenase require molybdopterin as a metal binding cofactor. Several genes necessary for the synthesis of the molybdenum cofactor (MoCo) have been characterized in bacteria and plants. The proteins encoded by the Escherichia coli genes moaA and moaC catalyse the first steps in MoCo synthesis. The human homologues of these genes are therefore candidate genes for molybdenum cofactor deficiency, a rare and fatal disease. Using oligonucleotides complementary to a conserved region in the moaA gene, we have isolated a human cDNA derived from liver mRNA. This transcript contains an open reading frame (ORF) encoding the human moaA homologue and a second ORF encoding a human moaC homologue. Mutations can be found in the majority of MoCo-deficient patients that confirm the functional role of both ORFs in the corresponding gene MOCS1 (for 'molybdenum cofactor synthesis-step 1'). Northern-blot analysis detected only full-length transcripts containing both consecutive ORFs in various human tissues. The mRNA structure suggests a translation reinitiation mechanism for the second ORF. These data indicate the existence of a eukaryotic mRNA, which as a single and uniform transcript guides the synthesis of two different enzymatic polypeptides with disease-causing potential. PMID- 9731532 TI - Sonic hedgehog is essential to foregut development. AB - Congenital malformation of the foregut is common in humans, with an estimated incidence of 1 in 3000 live births, although its aetiology remains largely unknown. Mice with a targeted deletion of Sonic hedgehog (Shh) have foregut defects that are apparent as early as embryonic day 9.5, when the tracheal diverticulum begins to outgrow. Homozygous Shh-null mutant mice show oesophageal atresia/stenosis, tracheo-oesophageal fistula and tracheal and lung anomalies, features similar to those observed in humans with foregut defects. The lung mesenchyme shows enhanced cell death, decreased cell proliferation and downregulation of Shh target genes. These results indicate that Shh is required for the growth and differentiation of the oesophagus, trachea and lung, and suggest that mutations in SHH and its signalling components may be involved in foregut defects in humans. PMID- 9731531 TI - Essential function of Gli2 and Gli3 in the formation of lung, trachea and oesophagus. AB - Foregut malformations (oesophageal atresia, tracheo-oesophageal fistula, lung anomalies and congenital stenosis of the oesophagus and trachea) are relatively common anomalies occurring in 1 in 2,000-5,000 live births, although their aetiology is poorly understood. The secreted glycoprotein Sonic hedgehog (Shh) has been suggested to act as an endodermal signal that controls hindgut patterning and lung growth. In mice, three zinc-finger transcription factors, Gli1, Gli2 and Gli3, have been implicated in the transduction of Shh signal. We report here that mutant mice lacking Gli2 function exhibit foregut defects, including stenosis of the oesophagus and trachea, as well as hypoplasia and lobulation defects of the lung. A reduction of 50% in the gene dosage of Gli3 in a Gli2-/- background resulted in oesophageal atresia with tracheo-oesophageal fistula and a severe lung phenotype. Mutant mice lacking both Gli2 and Gli3 function did not form oesophagus, trachea and lung. These results indicate that Gli2 and Gli3 possess specific and overlapping functions in Shh signalling during foregut development, and suggest that mutations in GLI genes may be involved in human foregut malformations. PMID- 9731533 TI - The APCI1307K allele and cancer risk in a community-based study of Ashkenazi Jews. AB - Mutations in APC are classically associated with familial adenomatous polyposis (FAP), a highly penetrant autosomal dominant disorder characterized by multiple intestinal polyps and, without surgical intervention, the development of colorectal cancer (CRC). APC is a tumour-suppressor gene, and somatic loss occurs in tumours. The germline T-to-A transversion responsible for the APC I1307K allele converts the wild-type sequence to a homopolymer tract (A8) that is genetically unstable and prone to somatic mutation. The I1307K allele was found in 6.1% of unselected Ashkenazi Jews and higher proportions of Ashkenazim with family or personal histories of CRC (ref. 2). To evaluate the role of I1307K in cancer, we genotyped 5,081 Ashkenazi volunteers in a community survey. Risk of developing colorectal, breast and other cancers were compared between genotyped I1307K carriers and non-carriers and their first-degree relatives. PMID- 9731534 TI - Trisomy 7-harbouring non-random duplication of the mutant MET allele in hereditary papillary renal carcinomas. AB - The gene defect for hereditary papillary renal carcinoma (HPRC) has recently been mapped to chromosome 7q, and germline mutations of MET (also known as c-met) at 7q31 have been detected in patients with HPRC (ref. 2). Tumours from these patients commonly show trisomy of chromosome 7 when analysed by cytogenetic studies and comparative genomic hybridization (CGH). However, the relationship between trisomy 7 and MET germline mutations is not clear. We studied 16 renal tumours from two patients with documented germline mutations in exon 16 of MET. Fluorescent in situ hybridization (FISH) analysis showed trisomy 7 in all tumours. To determine whether the chromosome bearing the mutant or wild-type MET gene was duplicated, we performed duplex PCR and phosphoimage densitometry using polymorphic microsatellite markers D7S1801 and D7S1822, which were linked to the disease gene locus, and D1S1646 as an internal control. We determined the parental origin of chromosome alleles by genotyping parental DNA. In all 16 tumours there was an increased signal intensity (2:1 ratio) of the microsatellite allele from the chromosome bearing the mutant MET compared with the allele from the chromosome bearing the wild-type MET. Our study demonstrates a non-random duplication of the chromosome bearing the mutated MET in HPRC and implicates this event in tumorigenesis. PMID- 9731535 TI - Schizophrenia susceptibility loci on chromosomes 13q32 and 8p21. AB - Schizophrenia is a common disorder characterized by psychotic symptoms; diagnostic criteria have been established. Family, twin and adoption studies suggest that both genetic and environmental factors influence susceptibility (heritability is approximately 71%; ref. 2), however, little is known about the aetiology of schizophrenia. Clinical and family studies suggest aetiological heterogeneity. Previously, we reported that regions on chromosomes 22, 3 and 8 may be associated with susceptibility to schizophrenia, and collaborations provided some support for regions on chromosomes 8 and 22 (refs 9-13). We present here a genome-wide scan for schizophrenia susceptibility loci (SSL) using 452 microsatellite markers on 54 multiplex pedigrees. Non-parametric linkage (NPL) analysis provided significant evidence for an SSL on chromosome 13q32 (NPL score=4.18; P=0.00002), and suggestive evidence for another SSL on chromosome 8p21-22 (NPL=3.64; P=0.0001). Parametric linkage analysis provided additional support for these SSL. Linkage evidence at chromosome 8 is weaker than that at chromosome 13, so it is more probable that chromosome 8 may be a false positive linkage. Additional putative SSL were noted on chromosomes 14q13 (NPL=2.57; P=0.005), 7q11 (NPL=2.50, P=0.007) and 22q11 (NPL=2.42, P=0.009). Verification of suggestive SSL on chromosomes 13q and 8p was attempted in a follow-up sample of 51 multiplex pedigrees. This analysis confirmed the SSL in 13q14-q33 (NPL=2.36, P=0.007) and supported the SSL in 8p22-p21 (NPL=1.95, P=0.023). PMID- 9731536 TI - HIRA, a mammalian homologue of Saccharomyces cerevisiae transcriptional co repressors, interacts with Pax3. AB - HIRA maps to the DiGeorge/velocardiofacial syndrome critical region (DGCR) at 22q11 (refs 1,2) and encodes a WD40 repeat protein similar to yeast Hir1p and Hir2p. These transcriptional co-repressors regulate cell cycle-dependent histone gene transcription, possibly by remodelling local chromatin structure. We report an interaction between HIRA and the transcription factor Pax3. Pax3 haploinsufficiency results in the mouse splotch and human Waardenburg syndrome (WSI and WSIII) phenotypes. Mice homozygous for Pax3 mutations die in utero with a phenocopy of DGS, or neonatally with neural tube defects. HIRA was also found to interact with core histones. Thus, altered stoichiometry of complexes containing HIRA may be important for the development of structures affected in WS and DGS. PMID- 9731537 TI - Targeted disruption of the biglycan gene leads to an osteoporosis-like phenotype in mice. AB - The resilience and strength of bone is due to the orderly mineralization of a specialized extracellular matrix (ECM) composed of type I collagen (90%) and a host of non-collagenous proteins that are, in general, also found in other tissues. Biglycan (encoded by the gene Bgn) is an ECM proteoglycan that is enriched in bone and other non-skeletal connective tissues. In vitro studies indicate that Bgn may function in connective tissue metabolism by binding to collagen fibrils and TGF-beta (refs 5,6), and may promote neuronal survival. To study the role of Bgn in vivo, we generated Bgn-deficient mice. Although apparently normal at birth, these mice display a phenotype characterized by a reduced growth rate and decreased bone mass due to the absence of Bgn. To our knowledge, this is the first report in which deficiency of a non-collagenous ECM protein leads to a skeletal phenotype that is marked by low bone mass that becomes more obvious with age. These mice may serve as an animal model to study the role of ECM proteins in osteoporosis. PMID- 9731538 TI - Identification of a candidate modifying gene for spinal muscular atrophy by comparative genomics. AB - Spinal muscular atrophy (SMA) is a common recessive disorder characterized by the loss of lower motor neurons in the spinal cord. The disease has been classified into three types based on age of onset and severity. SMA I-III all map to chromosome 5q13 (refs 2,3), and nearly all patients display deletions or gene conversions of the survival motor neuron (SMN1) gene. Some correlation has been established between SMN protein levels and disease course; nevertheless, the genetic basis for SMA phenotypic variability remains unclear, and it has been postulated that the loss of an additional modifying factor contributes to the severity of type I SMA. Using comparative genomics to screen for such a factor among evolutionarily conserved sequences between mouse and human, we have identified a novel transcript, H4F5, which lies closer to SMN1 than any previously identified gene in the region. A multi-copy microsatellite marker that is deleted in more than 90% of type I SMA chromosomes is embedded in an intron of this gene, indicating that H4F5 is also highly deleted in type I SMA chromosomes, and thus is a candidate phenotypic modifier for SMA. PMID- 9731539 TI - Sensory ataxia and muscle spindle agenesis in mice lacking the transcription factor Egr3. AB - Muscle spindles are skeletal muscle sensory organs that provide axial and limb position information (proprioception) to the central nervous system. Spindles consist of encapsulated muscle fibers (intrafusal fibers) that are innervated by specialized motor and sensory axons. Although the molecular mechanisms involved in spindle ontogeny are poorly understood, the innervation of a subset of developing myotubes (type I) by peripheral sensory afferents (group Ia) is a critical event for inducing intrafusal fiber differentiation and subsequent spindle formation. The Egr family of zinc-finger transcription factors, whose members include Egr1 (NGFI-A), Egr2 (Krox-20), Egr3 and Egr4 (NGFI-C), are thought to regulate critical genetic programs involved in cellular growth and differentiation (refs 4-8, and W.G.T. et al., manuscript submitted). Mice deficient in Egr3 were generated by gene targeting and had gait ataxia, increased frequency of perinatal mortality, scoliosis, resting tremors and ptosis. Although extrafusal skeletal muscle fibers appeared normal, Egr3-deficient animals lacked muscle spindles, a finding that is consistent with their profound gait ataxia. Egr3 was highly expressed in developing muscle spindles, but not in Ia afferent neurons or their terminals during developmental periods that coincided with the induction of spindle morphogenesis by sensory afferent axons. These results indicate that type I myotubes are dependent upon Egr3-mediated transcription for proper spindle development. PMID- 9731541 TI - Assignment of Tangier disease to chromosome 9q31 by a graphical linkage exclusion strategy. AB - A low level of high density lipoprotein (HDL) cholesterol is a strong predictor of ischaemic heart disease (IHD) and myocardial infarction. One cause of low HDL cholesterol is Tangier disease (TD), an autosomal codominant inherited condition first described in 1961 in two siblings on Tangier Island in the United States of America. Apart from low HDL-cholesterol levels and an increased incidence of atherosclerosis, TD is characterized by reduced total cholesterol, raised triglycerides, peripheral neuropathy and accumulation of cholesteryl esters in macrophages, which causes enlargement of the liver, spleen and tonsils. In contrast to two other monogenic HDL deficiencies in which defects in the plasma proteins apoA-I and LCAT interfere primarily with the formation of HDL (refs 7 10), TD shows a defect in cell signalling and the mobilization of cellular lipids. The genetic defect in TD is unknown, and identification of the Tangier gene will contribute to the understanding of this intracellular pathway and of HDL metabolism and its link with IHD. We report here the localization of the genetic defect in TD to chromosome 9q31, using a genome-wide graphical linkage exclusion strategy in one pedigree, complemented by classical lod score calculations at this region in a total of three pedigrees (combined lod 10.05 at D9S1784). We also provide evidence that TD may be due to a loss-of-function defect. PMID- 9731540 TI - A missense mutation in the alphaB-crystallin chaperone gene causes a desmin related myopathy. AB - Desmin-related myopathies (DRM) are inherited neuromuscular disorders characterized by adult onset and delayed accumulation of aggregates of desmin, a protein belonging to the type III intermediate filament family, in the sarcoplasma of skeletal and cardiac muscles. In this paper, we have mapped the locus for DRM in a large French pedigree to a 26-cM interval in chromosome 11q21 23. This region contains the alphaB-crystallin gene (CRYAB), a candidate gene encoding a 20-kD protein that is abundant in lens and is also present in a number of non-ocular tissues, including cardiac and skeletal muscle. AlphaB-crystallin is a member of the small heat shock protein (shsp) family and possesses molecular chaperone activity. We identified an R120G missense mutation in CRYAB that co segregates with the disease phenotype in this family. Muscle cell lines transfected with the mutant CRYAB cDNA showed intracellular aggregates that contain both desmin and alphaB-crystallin as observed in muscle fibers from DRM patients. These results are the first to identify a defect in a molecular chaperone as a cause for an inherited human muscle disorder. PMID- 9731542 TI - Introduction. Alzheimer's disease: from research to practice. PMID- 9731543 TI - Neuroimaging for diagnosis of dementia. AB - Although many clinicians consider neuroimaging studies as optional for the differential diagnosis of dementia, clinical experience suggests that they can improve diagnostic accuracy. Data are limited, however, on sensitivity, specificity, and cost-effectiveness of various neuroimaging techniques. The author reviews advantages and disadvantages of neuroimaging techniques for the differential diagnosis of dementia and describes strategies used for early detection of Alzheimer's disease, including combining positron emission tomography scanning with genetic risk assessment. Such approaches could provide a means for in vivo therapeutic monitoring of brain function during experimental antidementia treatment trials. PMID- 9731544 TI - New therapeutic approaches to cognitive impairment. AB - Therapeutic approaches to the cognitive impairment of dementia are making their way into clinical practice. Clinical pharmacologic approaches toward improvement of cognitive symptoms are discussed, with an emphasis on cholinergic approaches, since they currently appear most promising and since several cholinesterase inhibitors may soon be available for prescribing. As more knowledge is gained about dosing, side effects, and mechanisms of action, these drugs can be prescribed more efficiently. Current research approaches to slowing the rate of cognitive decline are discussed, including the use of antioxidants, monoamine oxidase-B inhibitors, and cholinesterase inhibitors. Drugs that improve cognition may also have effects on behavioral symptoms, severe dementia, and non Alzheimer's dementia. Evidence suggests that some dementia patients may be particularly responsive to intervention and that other medications may enhance response. Psychosocial interventions may also contribute to prolonging the time to institutionalization. PMID- 9731546 TI - Developing Treatment Guidelines for Alzheimer's Disease and Other Dementias. AB - Escalating health care costs and evidence that there is widespread variation in medical practice have led to the formation of more than 1800 consensus conferences in the past 10 years. These conferences seek to review the evidence that existing treatments have demonstrable efficacy; to determine if evidence favors one form of therapy over another; to review the guidelines for implementation, continuation, and discontinuation of the therapy; and to identify currently used treatments for which no benefit can be documented. This is a review of the process used by the American Psychiatric Association Task Force on Developing Treatment Guidelines for Alzheimer's Disease and Other Dementias, still in process at the time of this presentation. PMID- 9731545 TI - Future therapeutic approaches to Alzheimer's disease. AB - As palliative treatments for Alzheimer's disease proliferate, the focus of therapeutics turns to drugs with the potential to alter course. Evidence is reviewed, suggesting that there is biological plausibility to utilizing anti inflammatory agents, antioxidants, free-radical scavengers, estrogen preparations, and perhaps cholinomimetics. This range of possibilities leads to an optimistic assessment of the likelihood for altering the course or delaying the onset of Alzheimer's disease. PMID- 9731547 TI - Suppression of proliferative cholangitis in a rat model with direct adenovirus mediated retinoblastoma gene transfer to the biliary tract. AB - Proliferative cholangitis (PC) associated with hepatolithiasis develops the stricture of main bile ducts, and is the main cause of residual and/or recurrent stones after repeated treatments for hepatolithiasis. The aim of this study was to inhibit PC using the cytostatic gene therapy with direct adenovirus-mediated retinoblastoma (Rb) gene transfer to the biliary tract. PC was induced by introducing a fine nylon thread into the bile duct in a rat model. The adenovirus vector encoding a nonphosphorylatable, constitutively active form of retinoblastoma gene product (AdRb) was administered directly into the biliary tract. The adenovirus vector encoding beta-galactosidase (AdlacZ) was also given as a control. The bile duct wall thickness and 5'-bromodeoxyuridine (BrdU) labeling index were compared among uninfected, AdlacZ-infected, and AdRb-infected PC rats. The Rb expression in the bile duct was detected using reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemical study. AdRb-infected bile ducts showed inhibition of the epithelial and fibrous tissue proliferation and the peribiliary gland hyperplasia, resulting in a significant reduction of wall thickness compared with uninfected and AdlacZ-infected ones. The BrdU labeling index was 4.87% +/- 3.06% in the AdRb-infected bile ducts, while those of uninfected and AdlacZ-infected ones were 15.48% +/- 4.61% and 11.72% +/- 1.23%, respectively (P < .05). In conclusion, our cytostatic gene therapy approach using direct Rb gene transfer into the biliary tract suppressed PC in a rat model and may offer an effective therapeutic option for reducing recurrences following treatments against hepatolithiasis. PMID- 9731548 TI - Effect of the prokinetic agent, erythromycin, in the Richardson ground squirrel model of cholesterol gallstone disease. AB - Impaired gallbladder motility and delayed intestinal transit contribute to cholesterol gallstone formation by impeding the enterohepatic circulation of bile salts and causing gallbladder stasis. The therapeutic value of erythromycin, a prokinetic motilin analog, was evaluated in an animal model of gallstone formation. Eighty ground squirrels were fed either a trace- (control) or a high- (1%) cholesterol diet. Half of each diet group received either erythromycin stearate or placebo orally twice daily for 4 weeks. Biliary lipid secretion and bile salt pool size were determined via common duct cannulation. Gallbladder contractile response to cholecystokinin (CCK) was studied in vitro. Intestinal transit was evaluated in vivo by 51Cr marker. In the placebo-treated group, fed the high- versus the trace-cholesterol diet, bile salt secretion decreased (trace cholesterol + placebo, 21.0 +/- 1.8 nmol/min/g liver vs. high-cholesterol + placebo, 9.3 +/- 1.4 nmol/min/g liver), cholesterol saturation index (CSI) doubled (trace-cholesterol + placebo, 0.61 +/- 0.06 vs. high-cholesterol + placebo, 1.30 +/- 0.04), nucleation time shortened (trace-cholesterol + placebo, > 21 days vs. high-cholesterol + placebo, 6.4 +/- 1.0 days), cholesterol crystals formed, gallbladder contractility diminished, and intestinal transit was delayed (each P < .05). Erythromycin treatment of animals on the high-cholesterol diet restored gallbladder contractility and intestinal transit to control levels, increased bile salt secretion, reduced the total bile salt pool, lowered the cholesterol saturation of bile, lengthened the nucleation time, and so reduced crystal formation (each P < .05). Erythromycin enhances gallbladder motility and hastens intestinal transit, promoting more rapid enterohepatic cycling of bile salts. This increases bile salt secretion, improves cholesterol solubility, and reduces crystal development. PMID- 9731549 TI - Increased CD1d expression on small bile duct epithelium and epithelioid granuloma in livers in primary biliary cirrhosis. AB - Cluster of differentiation 1 (CD1) is a family of four distinct nonpolymorphic major histocompatibility complex class I-like molecules that can present microbial nonpeptide lipid antigens to T cells. Among the CD1 gene family, CD1d is found in a wide range of tissues including the intestine and liver, and has been proposed to play an important role in mucosal immunity. Primary biliary cirrhosis (PBC) is an immune-mediated liver disease involving the intrahepatic small bile ducts, which also belong to the mucosal immune system. In this study, we studied the expression of CD1d in patients with PBC and compared the data with those of patients with hepatic sarcoidosis, primary sclerosing cholangitis (PSC), chronic viral hepatitis (CVH), and normal liver as controls. CD1d was found to be expressed in hepatocytes in all cases examined, and in epithelioid granuloma cells in 19 of 22 PBC livers and in 4 of 4 livers with hepatic sarcoidosis. In addition, CD1d was focally expressed on epithelial cells of the small bile ducts in approximately 50% of the PBC patients but in no controls. Such bile duct epithelial staining of CD1d was seen in early-stage PBC and virtually absent in late-stage PBC. Moreover, there was no evidence of expression of CD1d in large bile duct epithelial cells of PBC. The CD1d on biliary epithelial cells in PBC may be involved in the antigen presentation of microbial lipid antigen(s) to surrounding T cells. Alternatively, modified endogenous lipidic compounds may share analogy with bacterial lipid antigens and explain CD1d expression, a possible epiphenomenon rather than a proof of bacterial involvement. PMID- 9731550 TI - Effects of bile duct ligation on hepatic expression of female-specific CYP2C12 in male and female rats. AB - Gender differences in hepatic sex steroid and drug metabolism result from hormonal regulation of specific cytochrome P450 genes (CYP). In male rats, bile duct ligation (BDL) is associated with down-regulation of the male-specific genes, CYP2C11 and CYP3A2, together with a decrease in serum testosterone levels and a two- to three-fold increase in serum estradiol concentrations. We anticipated that if estrogen is responsible for down-regulation of male-specific CYPs in BDL male rats, the female-specific CYP2C12, which is not normally present in adult male rat liver, should be up-regulated. We examined this proposal by determining the profile of hepatic cytochrome P450 enzymes in female rats subjected to BDL, and by seeking evidence for expression of CYP2C12 in male rats that do not normally express this gene. In female rats killed 5 days after BDL, total cytochrome P450 content and NADPH-cytochrome P450-reductase (P450 reductase) were decreased to 74% and 58% of control, respectively. Microsomal enzyme activities attributable to CYP2A1, CYP2C6, and CYP2E1 were 50% to 60% of control, but ethylmorphine N-demethylase, which in female liver is catalyzed by CYP2C12 and to a lesser extent CYP2C6, was significantly less affected (81% of control). Likewise, levels of CYP2C6 and P450-reductase proteins were decreased in proportion to the corresponding enzyme activities (50% to 60%), while CYP2C12 protein (and mRNA levels) were not altered in BDL female rat liver. In sham operated male rats, transcripts for CYP2C12 were rarely detected, but mRNA levels rose to appreciable levels within 24 hours of BDL, and CYP2C12 protein was expressed in hepatic microsomes of BDL male rats. Administration of estradiol to male rats produced a similar elevation of CYP2C12 mRNA, to values approximately 40% of female rats. It is concluded that CYP2C12 is up-regulated in male rats with cholestasis caused by BDL, while CYP2C12 protein is preserved in female rats when other microsomal proteins are decreased. These changes may be related to the increase in serum estradiol levels that result from altered hepatic steroid metabolism. The results demonstrate that activities of individual drug metabolizing enzymes in liver disease can be determined by dysregulation of the constitutive expression of hepatic CYP genes. PMID- 9731551 TI - Determinants of the selection of phosphatidylcholine molecular species for secretion into bile in the rat. AB - Certain phosphatidylcholine (PC) molecular species appear to be secreted into bile preferentially, but the mechanism for this selection remains obscure. We used multivariate analysis to examine the relationship between PC structure and the odds of secretion for individual PC species secreted into bile. PC was isolated from Folch extracts of bile and liver from rats, and individual molecular species of PC were quantified with reverse-phase high-performance liquid chromatography (HPLC). The odds of secretion for a given PC species were quantified as the ratio of its mole% in bile/mole% in liver. Regression analysis indicated that the odds of secretion were significantly related to length of both the sn-1 and sn-2 acyl chains (P < .0001 for both) and to relative hydrophobicity as determined by reverse-phase HPLC (P < .0001). In addition, the relationship between odds of secretion and sn-1 chain length was best described by a parabolic function. Considered together, these characteristics accounted for 88% of the observed differences in odds of secretion. This relationship between PC structure and odds of secretion was strikingly similar to the relationship between PC structure and affinity for bovine PC transfer protein. When multivariate models were used to predict both the odds of secretion and the affinity for PC transfer protein for a set of biologically plausible PC species, there was a linear relationship between the two. The likelihood of a given PC species being secreted into bile can be related to the structural characteristics of the acyl chains without having to postulate the existence of a special pool of PC destined for biliary secretion. Second, the structural characteristics that dictate selection of PC species for secretion into bile are similar to those that determine binding affinity for PC transfer protein, suggesting that the likelihood of a PC being secreted into bile is, in fact, closely related to its binding affinity for PC transfer protein (PC-TP). PMID- 9731552 TI - Acute hepatic allograft rejection: incidence, risk factors, and impact on outcome. AB - Hepatic allograft rejection remains an important problem following liver transplantation, and, indeed, complications related to the administration of immunosuppressive therapy remain a predominant cause of posttransplantation morbidity and mortality. The Liver Transplantation Database (LTD) was used to study a cohort of 762 consecutive adult liver transplantation recipients and determined the incidence, timing, and risk factors for acute rejection. We also evaluated the impact of histological severity of rejection on the need for additional immunosuppressive therapy and on patient and graft survival. Four hundred ninety (64%) of the 762 adult liver transplantation recipients developed at least one episode of rejection during a median follow-up period of 1,042 days (range, 336-1,896 days), most of which occurred during the first 6 weeks after transplantation. Multivariate analysis revealed that recipient age, serum creatinine, aspartate transaminase (AST) level, presence of edema, donor/recipient HLA-DR mismatch, cold ischemic time, and donor age were independently associated with the time to acute rejection. An interesting observation was that the histological severity of rejection was an important prognosticator: the use of antilymphocyte preparations was higher, and the time to death or retransplantation was shorter, for patients with severe rejection. Findings from this study will assist in decision-making for the use of immunosuppressive regimens and call into question whether complete elimination of all rejection or alloreactivity is a desirable goal in liver transplantation. PMID- 9731553 TI - Vasopressin reverses mesenteric hyperemia and vasoconstrictor hyporesponsiveness in anesthetized portal hypertensive rats. AB - We recently reported that vasopressin analogues correct the in vitro vascular hyporeactivity to adrenergic vasoconstrictors in portal hypertensive rats. The aim of the present study was to determine whether vasopressin reduces splanchnic blood flow in portal vein-ligated (PVL) rats by restoring vasoconstrictor responsiveness in vivo. The ultrasonic transit time-shift technique was used for blood flow measurements. At basal conditions, blood flow through the superior mesenteric artery was elevated 1.6-fold in PVL rats as compared with sham operated (SHAM) control rats. PVL rats also exhibited blunted mesenteric constrictor responses to the adrenoceptor agonist, phenylephrine (0.03-1 micromol x min(-1) x kg(-1)). Terlipressin (2-20 microg x k(-1)) and arginine vasopressin (3-300 pmol x min(-1) x kg(-1)) dose-dependently reduced, and at the highest doses, even abolished, the difference in mesenteric blood flow (MBF) between PVL and SHAM rats. When expressed as percent changes relative to baseline, mesenteric arterial responses to terlipressin and arginine vasopressin were found to be enhanced in PVL rats as compared with SHAM rats. Moreover, pretreatment with terlipressin (20 microg x kg(-1)) reversed the mesenteric hyporesponsiveness to phenylephrine of PVL rats. These vasopressin effects were independent of the nitric oxide (NO) pathway, because they were not mimicked by inhibition of NO synthesis with N(G)-nitro-L-arginine methyl ester (L-NAME) (0.1-10 mg x kg(-1)). These data indicate that pharmacological doses of vasopressin reverse the splanchnic hyperemia by restoring the responsiveness to adrenergic vasoconstrictors in portal hypertensive rats. PMID- 9731554 TI - Beneficial hemodynamic effects of bosentan, a mixed ET(A) and ET(B) receptor antagonist, in portal hypertensive rats. AB - In patients with cirrhosis, the plasma level of endothelin, a potent vasoconstrictor peptide, is elevated, and endothelin plays a role in increased intrahepatic vascular resistance. Thus, the aim of this study was to evaluate the hemodynamic effects of bosentan, a mixed ET(A) and ET(B) endothelin receptor antagonist in three models of portal hypertension. In all groups of rats, endothelin (2 microg/kg intravenously) administration significantly increased intrahepatic vascular resistance. In rats with secondary biliary cirrhosis, bosentan (30 mg/kg) significantly reduced portal pressure from 14.6 +/- 1.2 to 12.1 +/- 0.6 mm Hg, while portal blood flow and cardiac output increased by 45% and 57%, respectively. Thus, hepatocollateral vascular resistance decreased significantly from 177 +/- 19 to 101 +/- 9 dyn x s x cm(-5) x 10(-3). Similar results were observed in rats with CCl4-induced cirrhosis. In isolated perfused cirrhotic rat livers, bosentan (1 to 100 micromol/L) had no significant effect on hepatic vascular resistance. In portal vein-stenosed rats, bosentan administration significantly decreased portal pressure from 13.1 +/- 0.6 to 11.4 +/- 0.5 mm Hg by reducing portosystemic vascular resistance, because bosentan had no effect on vascular resistance of normal rat liver. In conclusion, bosentan administration decreased portal pressure in vivo by reducing hepatocollateral vascular resistance in rats with cirrhosis. Thus, mixed endothelin receptor antagonists might be a new approach in the pharmacological treatment of portal hypertension. PMID- 9731555 TI - Human leukocyte antigen-C genes and susceptibility to primary sclerosing cholangitis. AB - Genetic susceptibility to primary sclerosing cholangitis (PSC) is associated with the extended HLA A1-B8-DR3 haplotype and also with the DRB3*0101-DRB1*0301 DQA1*0103-DQB1*0603 haplotype. However, very few studies have considered the role of HLA C which lies between HLA A and B, is highly polymorphic, and encodes proteins which play an important role in immunoregulation and in disease susceptibility. Traditional assignment of HLA Cw antigens by serology is both inaccurate and unreliable, with a high error rate. The aim of this study was to characterize the distribution of HLA C alleles in a large group of patients with primary sclerosing cholangitis by using a recently developed polymerase chain reaction-based genotyping technique. Ninety-three white adult patients of northern European origin with well characterized PSC and 100 geographically and racially matched controls were studied. HLA C and HLA DRB1 alleles were assigned by polymerase chain reaction-based genotyping, HLA A and B antigens by standard microlymphocytotoxicity test and extended haplotypes were constructed according to known patterns of linkage disequilibrium. The Cw*07 gene was found in 67.7% of patients versus 54% of controls (P = .051, OR = 1.79). This increase was a result of inheritance of the Cw*0701 allele which was found in 51.6% of patients compared with 34% of controls (P = .013, OR = 2.07). There were no significant differences in the frequencies of any of the other Cw alleles including the Cw*07 group: Cw*0702, Cw*0703, and Cw*0704. HLA-encoded genetic susceptibility to PSC is associated with the HLA Cw*0701 allele, but the association is weak and may simply reflect linkage disequilibrium with the HLA B8-DR3 haplotype. These findings indicate that the telomeric limit of HLA-encoded susceptibility to primary sclerosing cholangitis lies close to the HLA C locus. PMID- 9731556 TI - Effects of protein kinase C modulators on Na+/K+ adenosine triphosphatase activity and phosphorylation in aortae from rats with cirrhosis. AB - Protein kinase C (PKC) modulates the activity and phosphorylation of the catalytic alpha-subunit of sodium-potassium-adenosine triphosphatase (Na+/K+ ATPase) in normal arteries. Because PKC is altered in cirrhotic aortae, Na+/K+ ATPase may also be altered in these arteries. The aim of the present study was to investigate alpha-subunit activity and phosphorylation in aortae from normal and cirrhotic rats, under baseline conditions and during exposure to PKC modulators. Alpha-subunit activity was assessed by measuring the amount of 32P released by hydrolysis of [gamma-32P]ATP in freshly isolated cell membranes (in the absence of PKC modulators only) and membrane depolarization caused by ouabain-induced alpha-subunit inhibition in isolated aortae (in the absence and presence of PKC modulators). Alpha-subunit phosphorylation was assessed by incorporation of 32P into alpha-subunits. Staurosporine, a PKC inhibitor, and phorbol 12,13-dibutyrate (PDBU), a PKC activator, were used. In addition, alpha-subunit expression was studied by Western blot analysis. In the absence of PKC modulators, the amount of 32P released by hydrolysis of [gamma-32P]ATP and ouabain-induced membrane depolarization were significantly lower in cirrhotic than in normal aortae. Staurosporine suppressed ouabain-induced membrane depolarization in cirrhotic and normal arteries. Ouabain-induced membrane depolarization was similar in cirrhotic aortae exposed to PDBU and in normal arteries studied under baseline conditions. Alpha-subunit phosphorylation was significantly lower in cirrhotic than in normal aortae, in aortae under baseline conditions, and in arteries exposed to staurosporine. Phosphorylation of the alpha-subunit was similar in cirrhotic aortae exposed to PDBU and in normal arteries under baseline conditions. Western blot analysis showed that the amount of alpha-subunit did not significantly differ between cirrhotic and normal aortae. In conclusion, a decrease in baseline Na+/K+ ATPase alpha-subunit activity occurs in aortae from cirrhotic rats as a result of reduced basal PKC activity. This PKC-dependent decreased alpha-subunit activity may be caused by a reduction in PKC-induced alpha-subunit phosphorylation. PMID- 9731557 TI - Neuropeptide Y stimulates bile secretion via Y1 receptor in the left dorsal vagal complex in rats. AB - Neuropeptide Y (NPY) injected into the cerebrospinal fluid and the left dorsal vagal complex enhances bile acid-independent and bicarbonate-dependent bile secretion through vagal muscarinic pathways in animal models. NPY binds to and activates six different receptor subtypes, and NPY Y1 and Y2 receptors are distributed in the dorsal vagal complex. We sought to determine which NPY receptor subtypes are involved in central stimulation of bile secretion by examining the effect of microinjection of specific NPY receptor agonists into the dorsal vagal complex. The bile duct was cannulated in urethane-anesthetized and bile acid-compensated rats. After measuring basal secretion, NPY, peptide YY (PYY), [Leu31, Pro34]NPY, NPY(13-36), or NPY(3-36) was microinjected into the either right or left dorsal vagal complex and bile secretion was observed for 100 minutes. Hepatic branch vagotomy was performed 2 hours before the peptide injection. Microinjection of NPY and PYY (8 pmol) into the left dorsal vagal complex increased bile secretion. [Leu31, Pro34]NPY microinjected into the left dorsal vagal complex also dose-dependently (1-8 pmol) stimulated bile acid independent and bicarbonate-dependent bile secretion. Microinjection of NPY(13 36) into the left dorsal vagal complex did not stimulate and NPY(3-36) dose dependently inhibited bile secretion. Stimulation of bile secretion by [Leu31, Pro34]NPY was abolished by hepatic branch vagotomy. NPY acts in the left dorsal vagal complex to stimulate bile acid-independent and bicarbonate-dependent bile secretion via Y1 receptor subtype. PMID- 9731558 TI - Effects of propranolol on the hepatic hemodynamic response to physical exercise in patients with cirrhosis. AB - Physical exercise increases portal pressure (hepatic venous pressure gradient [HVPG]) in patients with cirrhosis. It is unknown if this deleterious effect is associated with changes in gastroesophageal collateral blood flow and if these can be prevented by propranolol administration. The aim of this study was to characterize the effects of propranolol on the splanchnic hemodynamic response to exercise in patients with cirrhosis. Twenty-three patients with cirrhosis and portal hypertension had hemodynamic measurements in baseline conditions, and during moderate cycling exercise (40 W) under double-blind propranolol or placebo administration. In patients receiving placebo, HVPG significantly increased during exercise (from 16.7 +/- 0.9 to 19.0 +/- 1.0 mm Hg; P < .01), hepatic blood flow (HBF) decreased (-18% +/- 4%; P < .01), while azygos blood flow (AzBF) was unchanged (4% +/- 12%; ns). In patients receiving propranolol, portal pressure did not increase during exercise, but decreased from 16.3 +/- 1.0 to 12.9 +/- 1.1 mm Hg (P < .01). The lack of increase in HVPG in response to exercise in patients receiving propranolol may be related to a more pronounced decrease in HBF, as compared with patients receiving placebo, and to a blunted increase in cardiac output (CO). Moderate physical exercise adversely influences the hepatic hemodynamics in patients with cirrhosis, causing a significant increase in portal pressure. This is effectively prevented by propranolol pretreatment. PMID- 9731559 TI - Renal effects of transjugular intrahepatic portosystemic shunt in cirrhosis: comparison of patients with ascites, with refractory ascites, or without ascites. AB - Renal effects of the transjugular intrahepatic portosystemic shunt (TIPS) were compared in 6 patients without ascites (group 1), 11 patients with ascites responding to diuretic treatment (group 2), and 6 patients with refractory ascites (group 3). Seven days after insertion of TIPS, 24-hour urinary sodium excretion had increased in patients with ascites: 113 +/- 16 mmol to 170 +/- 30 mmol (P = .012) in group 2, and 22 +/- 8 mmol to 77 +/- 27 mmol (P = .050) in group 3. In group 3, fractional sodium excretion tended to increase from 0.26% +/ 0.14% to 0.62% +/- 18% (P = .081). The relative increase of urinary sodium excretion (to 444% +/- 122%) and fractional sodium excretion (to 413% +/- 127%) in group 3 was significantly (P < .05) higher than in group 1 and group 2, respectively. Creatinine clearance and 24-hour urinary volume were not significantly changed in either group. Patients with Child-Pugh class C had a more pronounced effect of TIPS on urinary sodium excretion (increase to 396% +/- 115% vs. 139% +/- 15%; P = .066) and on fractional sodium excretion (increase to 415% +/- 103% vs. 94% +/- 15%; P = .020) than patients with less-severe liver disease. Fractional sodium excretion of less than 0.35% before TIPS was found to be an indicator of renal response to TIPS. The effect of TIPS on urinary sodium excretion and on fractional sodium excretion was related to the patients' Child Pugh score (r = .55; P = .007 and r = .68; P = .001, respectively) and inversely to their fractional sodium excretion (r = -.44; P = .047 and r = -.54; P = .012, respectively) before TIPS. These data demonstrate that TIPS affects renal sodium handling in patients with ascites, particularly in patients with refractory ascites. Severity of liver disease and fractional sodium excretion before TIPS are parameters to predict the extent of the renal response. PMID- 9731560 TI - N-acetylcysteine prevents development of the hyperdynamic circulation in the portal hypertensive rat. AB - Partial portal vein ligation (PPVL) leads to the development of a hyperdynamic circulation. It is associated with elevated levels of tumor necrosis factor (TNF alpha) and nitric oxide (NO) production, both of which can result in oxidant injury. In this study, we have investigated whether PPVL is associated with the development of oxidative stress, by measuring urinary F2-isoprostanes. In addition, we have examined whether N-acetylcysteine (NAC) can ameliorate oxidant injury and prevent the development of the hyperdynamic circulation. Urinary excretion of F2-isoprostanes increased sixfold following PPVL together with a significant increase in plasma nitrite and nitrate. Treatment with NAC inhibited the formation of F2-isoprostanes as well as the increase in plasma nitrite and nitrate. Hemodynamic studies in anesthetized rats showed that following PPVL, cardiac output and portal pressure increased, and systemic vascular resistance decreased, consistent with the development of a hyperdynamic circulation. These changes were prevented by chronic administration of NAC. We conclude that NAC prevents the development of the hyperdynamic circulation and that the formation of reactive oxygen species may be important in the pathogenesis of these hemodynamic changes. PMID- 9731561 TI - Bone mineral density, serum insulin-like growth factor I, and bone turnover markers in viral cirrhosis. AB - Previous studies suggest that low bone mass is a complication of alcoholic liver disease. Nevertheless, little is known about bone mass and bone metabolism in viral cirrhosis. To evaluate the prevalence and magnitude of hepatic osteopenia in these patients, bone remodeling status, and its relationship with the severity of liver disease and serum levels of insulin-like growth factor I (IGF-I), we studied 32 consecutive patients with viral cirrhosis and no history of alcohol intake. Bone mineral density (BMD) was measured by dual x-ray absorptiometry in the lumbar spine (LS) and femoral neck (FN), and the values were expressed as the z score. Bone metabolism markers and hormone profiles were measured. Patients with viral cirrhosis showed reduced BMD in all sites (LS: -1.27 +/- 1.06, P < .001; FN: -0.48 +/- 0.96; P < .01). Of the 32 patients, 53% met the diagnostic criteria for osteoporosis. In patients, urine deoxypyridinoline (D-Pyr) as a marker of bone resorption and serum bone alkaline phosphatase (b-AP) as a marker of bone formation were significantly higher than in control subjects (P < .001 and P < .01, respectively). Serum IGF-I was lower than in control subjects (P < .001), and significant differences were also found between patients with and without osteoporosis (P < .05). BMD in LS correlated with severity of the disease, with serum levels of IGF-I, and with urine D-Pyr. Our findings show that viral cirrhosis is a major cause of osteoporosis in men, and that low serum IGF-I levels seem to play a role in the bone mass loss in these patients. The biochemical markers of bone remodeling suggest high-turnover osteoporosis in patients with viral cirrhosis. PMID- 9731562 TI - Detection of cholangiocarcinoma in primary sclerosing cholangitis by positron emission tomography. AB - Primary sclerosing cholangitis (PSC) predisposes to cholangiocarcinoma (CC), which usually is widespread in the liver at the time of the diagnosis and which has a median survival of approximately 6 months. Positron emission tomography (PET) is a noninvasive scanning method that allows the assessment of metabolism in vivo by means of positron-emitting radiolabeled tracers. [18F]Fluoro-2-deoxy-D glucose (FDG) is a glucose analogue that accumulates in various malignant tumors because of their high glucose metabolic rates. The purpose of the study was to develop a PET method to detect small CC tumors in patients with PSC. PET scanning of the liver was performed after intravenous injection of 200 MBq FDG in 9 patients with PSC, 6 patients with PSC + CC, and 5 controls. The scanning was performed at successive time intervals for a total of 90 minutes with simultaneous successive arterial blood sampling for radioactivity concentration determination. In each of the PSC + CC patients, 2 to 7 "hot spots" were seen, with volumes of 1.0 to 45 mL (median, 4.4 mL). There were no hot spots in the two other patient groups. The localization of hot spots was confirmed by single-blind evaluation. Data were analyzed by the Gjedde-Patlak plot, yielding values of the net metabolic clearance of FDG, K [mL min(-1) 100 mL(-1) tissue]. In the CC hot spots, maximum K values were 1.59 to 4.17 (median, 2.34; n = 6); in the reference liver tissues of these patients, K values were 0.40 to 0.69 (median, 0.49); in PSC patients, they were 0.23 to 0.53 (median, 0.36); and in controls, they were 0.20 to 0.34 (median, 0.31). The difference between K in CC hot spots and the other groups was statistically significant (P < .001). We conclude that FDG-PET seems to be able to detect small CC tumors and may be useful in the therapeutic management of PSC. PMID- 9731563 TI - Hepatocyte growth factor induces hepatocyte proliferation in vivo and allows for efficient retroviral-mediated gene transfer in mice. AB - Recombinant retroviral vectors are an attractive means of transferring genes into the liver because they integrate into the host cell genome and result in permanent gene expression. However, efficient in vivo gene transfer is limited by the requirement of active cell division for integration. Traditional approaches to induce liver proliferation have the disadvantage of inducing hepatocellular injury by delivery of toxins or by surgical partial hepatectomy. As a nontraumatic alternative, we show that exogenous hepatocyte growth factor (HGF) is a powerful and safe mitogen for the mature intact murine liver when delivered continuously into the portal vein. A 5-day infusion of human HGF (5 mg/kg/d) resulted in > 140% increase in relative liver mass, which returned to normal in 4 to 5 weeks. This clearly shows that an exogenous growth factor can induce robust liver proliferation in vivo. In addition, we show that the HGF-induced proliferation was independent of interleukin-6, an essential cytokine involved in liver regeneration after partial hepatectomy. When recombinant retroviral vectors were infused in combination with HGF, 30% of hepatocytes were stably transduced with no indication of hepatic injury or histopathology. These results show the ability to obtain a clinically relevant transduction efficiency with retroviral vectors in vivo without the prior induction of liver injury. The level of hepatic gene transfer achieved has the potential to be curative for a large number of genetic liver diseases. PMID- 9731564 TI - Levels of transforming growth factor beta and transforming growth factor beta receptors in rat liver during growth, regression by apoptosis and neoplasia. AB - Transforming growth factor beta1 (TGF-beta1) has been implicated as inhibitor of cell proliferation and a potent inducer of apoptosis in vitro and in vivo after the administration of high doses. To assess the role of endogenous TGF-beta1, we quantitated the cytokine and its receptors in rat liver during regenerative and hyperplastic growth, regression by apoptosis, and in hepatocellular carcinoma (HCC). This was accomplished by Northern blot analysis and by RNase protection assay of the messenger RNA (mRNA) of TGF-beta1 and TGF-beta receptors (TbetaR) types I to III and by an activity bioassay of the TGF-beta proteins. Untreated rat livers were found to contain 15.6 +/- 4.8 ng TGF-beta1 protein/g tissue; TGF beta2 protein was not detected. To induce toxic cell death and subsequent regenerative DNA synthesis in the liver, rats were treated with a necrogenic dose of carbon tetrachloride (CCl4). After 24 and 48 hours, there was an upregulation of TGF-beta1 (mRNA, up to tenfold; protein, about twofold) and of TbetaRs (mRNA: two- to fourfold); that indicates an overall enhanced production of and sensitivity to TGF-beta1, which may serve to confine the regenerative response. Hyperplastic liver growth and regression of the hyperplasia were induced by treatment with cyproterone acetate (CPA) or nafenopin (NAF) followed by withdrawal; neither mRNAs of TGF-beta1 and TbetaR types I to III nor TGF-beta1 protein exhibited significant changes during the growth phase or during regression by apoptosis. We also studied neoplastic growth. HCC, obtained after long-term treatment with NAF, exhibited high rates of cell replication and apoptosis. The majority of lesions contained mRNA and protein of TGF-beta1 and mRNA of TbetaR types I to III at concentrations similar to those of the surrounding tissue. In conclusion, during liver regeneration there is a pronounced upregulation of expression of both TGF-beta1 and TbetaRs I to III, but not during mitogen-induced liver growth or regression. It appears that apoptosis is induced via altered local concentration of TGF-beta1, in a paracrine and/or autocrine way. By this mechanism the lethal effects of TGF-beta1 may be locally confined, and overshoots of apoptosis in the liver may be prevented. PMID- 9731565 TI - Retinoic acid treatment induces apoptosis or expression of a more differentiated phenotype on different fractions of cultured fetal rat hepatocytes. AB - The present study reports the effects of retinoic acid (RA) on cultured fetal rat hepatocytes. We show that RA treatment induces both differentiation and apoptosis. Hepatocytes cultured for 48 hours in the presence of 5 microl/L RA form junctional complexes in the areas of contact between neighboring cells and develop bile canaliculi, typical features of mature and well-differentiated cells. At the same time, about 20% of cells are induced to die by apoptosis, and the percentage of apoptotic cells increases according to the concentration of RA used and the duration of treatment. The induction of apoptosis, studied at the morphological and biochemical levels, revealed that, in our system, the classical compaction of chromatin occurs only during the final stages of the process; instead of the common marker of apoptosis, i.e., the "DNA ladder" pattern of fragmentation, megabase-sized fragments were found. These observations provide further evidence of the existence of fundamental differences in the mechanisms of apoptosis among cell types. To investigate the molecular mechanism of the effects of RA, we evaluated the expression of two proteins, c-myc and p53, which are known to be involved in both cell differentiation and apoptosis. The data obtained show that the amount of p53 remained unchanged after RA treatment. On the contrary, a dose-dependent reduction in c-myc levels was found, suggesting that RA action may be mediated by modulation of this oncogene. Our findings regarding the apoptosis-inducing effect of RA, which was not found in adult hepatocytes, suggest a possible relationship between this phenomenon and the proliferative capacity and/or differentiation state of hepatocytes. PMID- 9731566 TI - Expression of cyclin-dependent kinase inhibitor p21 in human liver. AB - The p21 protein is a universal inhibitor of cyclin-dependent kinases and of cell cycle progression and is involved in numerous growth-inhibitory pathways in cell culture systems. Recent studies suggest that p21 regulates hepatocyte cell cycle progression in models of liver regeneration. The present study was designed to investigate the possible involvement of p21 in the control of hepatocyte proliferation in human liver diseases. To examine that, the expression of p21 in clinical liver biopsy specimens was determined by immunohistochemistry. This was correlated with hepatocyte Ki-67 immunostaining (a marker of hepatocyte proliferation in vivo) as well as histologic features. Little p21 or Ki-67 expression was detected in normal human liver or in specimens of nonalcoholic steatohepatitis. In patients with alcoholic hepatitis, increased expression of p21, but not of Ki-67, was observed. In specimens with chronic hepatitis C, hepatocyte p21 expression was significantly correlated with Ki-67 immunostaining, as well as with the degree of inflammation and fibrosis. These results indicate that hepatocyte p21 expression is upregulated in response to hepatic injury and correlates with histologic markers of proliferation and disease activity. This study provides evidence that p21 plays a role in the regulation of hepatocyte proliferation in human liver diseases. PMID- 9731567 TI - Chronic alcohol intake reduces retinoic acid concentration and enhances AP-1 (c Jun and c-Fos) expression in rat liver. AB - Chronic ethanol intake may interfere with retinoid signal transduction by inhibiting retinoic acid synthesis and by enhancing activator protein-1 (AP-1) (c Jun and c-Fos) expression, thereby contributing to malignant transformation. To determine the effect of ethanol on hepatic retinoid levels, retinoic acid receptors (RARs) and AP-1 (c-Jun and c-Fos) gene expression, chronic ethanol (36% of total calorie intake) pair-feeding was conducted on rats for a 1-month period. Retinoic acid, retinol, and retinyl ester concentrations in both liver and plasma were examined by using high-performance liquid chromatography (HPLC). Both retinoic acid receptor (alpha, beta, gamma) and AP-1 (c-Jun and c-Fos) expression in the rat liver were examined by using Western blot analysis. Treatment with high-dose ethanol led to a significant reduction of retinoic acid concentration in both the liver and the plasma (11- and 8.5-fold reduction, respectively), as compared with animals pair-fed an isocaloric control diet containing the same amount of vitamin A. Similar to the retinoic acid reductions, both retinol and retinyl palmitate levels in the livers of the alcohol-fed group decreased significantly, but in smaller fold reduction (6.5- and 2.6-fold reduction, respectively). Ethanol did not modulate the expression of RARalpha, -beta, and gamma genes in the liver. However, chronic alcohol feeding enhanced AP-1 (c-Jun and c-Fos) expression by 7- to 8-fold, as compared with the control group. These data suggest that functional downregulation of RARs by inhibiting biosynthesis of retinoic acid and up-regulation of AP-1 gene expression may be important mechanisms for causing malignant transformation by ethanol. PMID- 9731568 TI - A new prognostic system for hepatocellular carcinoma: a retrospective study of 435 patients: the Cancer of the Liver Italian Program (CLIP) investigators. AB - The clinical outcome of cirrhotic patients with hepatocellular carcinoma (HCC) depends both on the residual liver function and tumor characteristics. However, the relative prognostic weight of these variables is not well defined. The aims of this study were to verify the value of known prognostic factors and to devise a prognostic index more sensitive than the commonly used Okuda stage. A retrospective analysis of the cases of HCC diagnosed at 16 Italian institutions from 1990 to 1992 was performed. Overall survival was the only end point used in the analysis. The Cox model, stratified by locoregional treatment, was used for multivariate analyses. The final model was derived from 10 randomly chosen training samples, and the prognostic validity of the Cancer of the Liver Italian Program (CLIP) score was assessed on the corresponding testing samples. Four hundred thirty-five cases of HCC were collected. As of January 1997, 313 patients (72%) were deceased. Overall median survival was 20 months. At multivariate analysis, independent predictive factors of survival were Child-Pugh stage, tumor morphology, alpha-fetoprotein (AFP), and portal vein thrombosis. A simple scoring system (CLIP score) was thus produced, assigning linear scores (0/1/2) to the covariates. Compared with Okuda stage, the CLIP score, structured as a six category tool, has a greater discriminant ability, revealing a class of patients with an impressively more favorable prognosis and another class with a relatively shorter life expectancy. The CLIP score is a new prognostic system that accounts for both liver function and tumor characteristics. It is easy to calculate and appears to give more precise information than the Okuda stage. PMID- 9731569 TI - Portal branch ligation with a continuous hepatocyte growth factor supply makes extensive hepatectomy possible in cirrhotic rats. AB - In a cirrhotic liver, the regenerative ability and specific functions are so impaired that excessive resection easily complicates postoperative liver dysfunction, which frequently leads to life-threatening multiple-organ failure. Hepatocyte growth factor (HGF), first identified as the most potent stimulator of DNA synthesis in primary hepatocytes, not only stimulates liver regeneration, but also accelerates hepatic function, improves fibrosis, and protects liver cells against injury. Therefore, we investigated the efficacy of preoperative portal branch ligation (PBL) (which can induce compensatory hypertrophy of the unaffected lobes) combined with a continuous HGF supply in the performance of extensive hepatectomy in cirrhotic rats. Cirrhosis was induced by intraperitoneal injections of dimethylnitrosamine (DMN) three times per week for 3 weeks. Five days after the last injection, when 70% hepatectomy is lethal, the rats underwent portal ligation of the left lateral and median branches (corresponding to approximately 70% of the total volume of the liver). Simultaneously, they were continuously treated with either recombinant human HGF (rhHGF) or vehicle from an intraperitoneally implanted osmotic pump. Four days after the portal ligation, the occluded lobes were resected. The HGF treatment rapidly increased both the wet weight of the unoccluded lobes and the hepatocellular DNA synthesis. The blood chemical analysis indicated that HGF significantly suppressed the posthepatectomy liver dysfunction. Most importantly, the HGF treatment markedly improved the survival rate of the rats at 48 hours after the major hepatectomy. In conclusion, PBL combined with a continuous HGF supply makes extensive hepatectomy possible in cirrhotic rats, mainly by promoting the hypertrophy of the unaffected lobes. PMID- 9731570 TI - Parenchymal cell apoptosis as a signal for sinusoidal sequestration and transendothelial migration of neutrophils in murine models of endotoxin and Fas antibody-induced liver injury. AB - Endotoxin (ET) induces neutrophil sequestration in hepatic sinusoids, the activation of proinflammatory transcription factors (nuclear factor KB [NF kappaB]) with up-regulation of adhesion molecules on sinusoidal endothelial cells and hepatocytes. However, if galactosamine (Gal) is co-administered with ET, neutrophils transmigrate and attack parenchymal cells. This suggests that a signal from parenchymal cells triggers neutrophil transmigration. In this study, we tested the hypothesis that parenchymal cell apoptosis may induce neutrophil transendothelial migration in the Gal/ET model. Treatment of C3Heb/FeJ mice with 700 mg/kg Gal and 100 microg/kg ET induced tumor necrosis factor alpha (TNF alpha) formation (13.25 +/- 0.75 ng/mL) and hepatic NF-kappaB activation at 90 minutes; the generation of the C-X-C chemokine KC (2.86 +/- 0.30 ng/mL at 5 hours); sinusoidal neutrophil sequestration (380 +/- 21 polymorphonuclear leukocytes/50 high-power fields) and apoptosis (925% +/- 29% increase of DNA fragmentation; and a 45-fold increase of terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL)-positive cells) at 6 hours, followed by transmigration of neutrophils and development of substantial necrosis (38% +/- 3% of hepatocytes; alanine transaminase [ALT]: 1,500 +/- 300 U/L) at 7 hours. Administration of uridine (1,000 mg/kg) did not reduce plasma levels of TNF-alpha and KC, NF-kappaB activation, or polymorphonuclear leukocyte sequestration, but attenuated apoptosis by 90% to 94%. In these livers, neutrophils did not transmigrate and liver injury was prevented (necrosis: < 5%; ALT: 40 +/- 3 U/L). However, massive apoptosis and liver injury initiated by the anti-Fas antibody, Jo2, did not recruit neutrophils into the liver. We conclude that excessive parenchymal cell apoptosis represents an important signal for transmigration of primed neutrophils sequestered in sinusoids during endotoxemia in vivo. However, apoptosis per se does not cause neutrophil sequestration in the liver vasculature. PMID- 9731571 TI - Hepatic preconditioning in rats is defined by a balance of adenosine and xanthine. AB - The present work investigates the relationship between adenosine, nitric oxide (NO), and free radicals during ischemic preconditioning in rat liver. For this purpose, we evaluated: 1) the efficacy of different periods of preconditioning; 2) the changes in the concentration of adenine nucleotides during preconditioning; 3) the importance of adenosine and xanthine concentrations in the induction of preconditioning; and 4) the possible effect of xanthine oxidase derived superoxide anion on NO during preconditioning. Results show that just a 10- to 15-minute period of ischemia followed by 10-minute reperfusion prevents the ischemic damage that would be induced by a subsequent 90 minutes of ischemia followed by 90 minutes of reperfusion. Administration of xanthine or metabolization of endogenous adenosine abolishes the protective effect of preconditioning. When rats have been subjected to a period of preconditioning not within the effective time window (10-15 minutes), and thus offering no protection, the administration of a NO donor was found to restore the protection. The dose needed to restore protection appears to be proportional to the endogenous xanthine concentration. In addition, when xanthine oxidase was inhibited, preconditioning effectively offered protection in front of ischemia and reperfusion, independently of the xanthine concentration. Altogether, this indicates that the time window of ischemia capable to induce preconditioning in liver is defined by the relative tissue concentrations of adenosine and xanthine. The lower limit of this window (10 minutes) is defined by the amount of adenosine able to induce NO generation. Its upper limit (15 minutes) is defined by the concentration of xanthine able to remove the generated NO. PMID- 9731572 TI - Osmotic regulation of the heat shock response in primary rat hepatocytes. AB - The influence of cell hydration and taurine on the heat shock response was studied in primary rat hepatocytes. Heat-induced accumulation of inducible heat shock protein 70 (HSP70) mRNA and protein was increased under hypo-osmotic conditions. In contrast, hyper-osmotic exposure blocked the HSP70 response during an 8-hour recovery, and this was paralleled by a reduction of overall protein synthesis and an impairment of thermotolerance. Taurine counteracted the hyper osmotic inhibition of heat-induced HSP70 expression, but increased overall protein synthesis only slightly. A rapid and transient activation of the stress activated protein kinase, JNK-2, was triggered by hyper-osmolarity, whereas the JNK-2 response to hypo-osmolarity was delayed. JNK-2 activation in response to heat was suppressed by hypo-osmolarity, but was markedly increased under hyper osmotic conditions. The latter effect was blocked by taurine. A pronounced induction of the mRNA for the MAP-kinase phosphatase, MKP-1, in response to heat was observed during hypo- and normo-osmolarity, but no MKP-1 induction was found under hyper-osmotic conditions, although hyper-osmolarity itself led to accumulation of small levels of MKP-1 mRNA. Also, the block of heat-induced MKP-1 mRNA expression by hyper-osmolarity was abolished in the presence of taurine. The data provide evidence for a role of cellular hydration and taurine in the protection of liver parenchymal cells against heat injury via regulation of HSP70 expression and the balance between JNK-2 and MKP-1 activity. PMID- 9731573 TI - Modulation of endothelin and nitric oxide: a rational approach to improve canine hepatic microcirculation. AB - ET receptor blocker (TAK-044) and NO donor (FK409) were used to improve the hepatic microcirculation following ischemia-reperfusion injury. In the first experiment (60 minutes of ischemia), 15 dogs were divided into three groups: group A (control), saline; group B, TAK 5 mg/kg; and group C, FK 0.4 mg/kg. In the second experiment (90 minutes of ischemia), 38 dogs were divided into six groups that underwent 90 minutes of hepatic ischemia followed by reperfusion: group I (control), saline only; group II, TAK 5 mg/kg and FK 3.2 mg/kg; group III, TAK 5 mg/kg and FK 0.4 mg/kg; group IV, TAK 5 mg/kg; group V, FK 0.4 mg/kg; and group VI, FK3.2 mg/kg. All drugs were administered through the portal vein. Following 60 minutes of ischemia, both FK and TAK produced significant improvement in hepatic microcirculation and enzymatic status when compared with the control group. After 90 minutes of ischemia, low doses of FK and TAK significantly improved hepatic microcirculation and reduced portal pressure following reperfusion in group III compared with group I. Leakage of hepatic enzymes was prevented and tissue injury score was significantly lower in group III. In group VI, early protection was obtained to some extent; however, blood pressure was reduced significantly following reperfusion compared with group I. In contrast, hepatocellular function deteriorated and the tissue injury score was higher in group II animals. TAK pretreatment with low doses of FK provided the best protection for the hepatic microcirculation in ischemia-reperfusion injury of the liver. PMID- 9731574 TI - Maternal cholestasis does not affect the ontogenic pattern of expression of the Na+/taurocholate cotransporting polypeptide (ntcp) in the fetal and neonatal rat liver. AB - To assess the effects of cholestasis during pregnancy on fetal and neonatal mRNA expression, protein mass, and function of the Na+/taurocholate cotransporting polypeptide (Ntcp), common bile duct ligation (BDL) was performed in pregnant rats on day 14 of pregnancy (maternal cholestasis [MC] group), and livers were harvested at days 20 and 21 of fetal life, as well as at days 4, 7, 14, 21, and 28 after birth. Sham-operated rats and their litters were used as controls. Ntcp steady-state mRNA levels, protein mass, and function were determined by Northern blotting, immunoblotting, and taurocholate (TC) transport studies in isolated short-term cultured hepatocytes, respectively. In addition, protein mass and function of the organic anion transporting polypeptide (Oatp1), another sinusoidal bile acid transporter, were studied at 4 weeks of age. The majority of pregnant cholestatic rats (94%) were able to carry pregnancy to term. Body and liver weights of the offspring from the MC group were lower than those from sham operated animals at all postnatal time points. Ntcp steady-state mRNA levels, protein mass, and function were unaffected by MC. The ontogenic pattern of expression was identical in offspring from MC and controls with detection of the Ntcp mRNA at day 21 of fetal life. There was a significant increase in mRNA postnatally, reaching adult levels by 7 days of age. Protein mass and function of Ntcp as well as of Oatp1 were similar in offspring from MC and control groups at 4 weeks of age. In conclusion, maternal obstructive cholestasis during the last third of pregnancy does not affect the fetal/neonatal expression of the basolateral bile acid transporters, Ntcp and Oatp1. This suggests that the impaired bile acid excretion described in this experimental model is not related to altered uptake of bile acids in the affected neonate. PMID- 9731575 TI - Altered regulation of Src tyrosine kinase by transforming growth factor beta1 in a human hepatoma cell line. AB - Transforming growth factor betas (TGF-betas) are the potent growth inhibitors for various cell types. Certain transformed cells, however, show poor response to TGF beta-induced growth inhibition, which contributes to their uncontrolled proliferation. Recently, we have reported that TGF-beta1 induces degradation of activated Src tyrosine kinase in rat fibroblasts. To elucidate the alteration in TGF-beta signaling pathway in tumor cells that cannot respond to the cytokine, we compared the effects of TGF-beta1 on Src kinase in two human hepatoma cell lines, TGF-beta1-insensitive Mahlavu cells and TGF-beta1-sensitive HepG2 cells. TGF beta1 decreased Src kinase activity in HepG2 cells, but increased cellular Src levels and Src kinase activity in Mahlavu cells. Co-incubation of Mahlavu cells with TGF-beta1 and 12-O-tetradecanoyl phorbol 13-acetate (TPA) decreased Src protein levels and Src kinase activity, inducing TGF-beta1 sensitivity. TGF-beta1 induced tyrosine dephosphorylation of Ras guanosine triphosphatase-activating protein (Ras-GAP) and Ras inactivation in HepG2 cells, but induced Ras-GAP phosphorylation and Ras activation in Mahlavu cells. The Src kinase inhibitor abolished the increase of Src kinase activity in TGF-beta1-treated Mahlavu cells, and induced TGF-beta1 sensitivity. These findings suggest that regulation of Src kinase by TGF-beta1 is altered in Mahlavu cells. The altered regulation of Src may contribute to TGF-beta1 insensitivity in this cell line, at least in part through activation of Ras. PMID- 9731576 TI - Impact of alcohol on the histological and clinical progression of hepatitis C infection. AB - In patients infected with the hepatitis C virus (HCV), 20% to 30% will progress to cirrhosis in over two to three decades. Viral and host factors that are important in the clinical and histologic progression of HCV infection are not entirely certain. It has been suggested that liver disease is worse in alcoholics infected with HCV. In the present retrospective study, we examined the effect of moderate alcohol intake on the histologic and clinical progression of HCV infection and assessed whether other variables such as gender, length of exposure, mode of exposure, HCV RNA levels, and ferritin levels also independently impacted disease progression. Liver biopsies were analyzed for the degree of fibrosis, presence of cirrhosis, and histologic activity by using the Histologic Activity Index of Knodell. Patients were divided into two groups based on whether their alcohol intake was significant or not significant. Significant alcohol intake was defined as > 40 g alcohol/day in women and > 60 g of alcohol/day in men for > 5 years. Groups were further divided based on the decades of exposure to HCV. There was no difference in the age or length of exposure to HCV in the alcohol and the alcohol-free group. HCV RNA serum levels, ferritin levels, and viral genotypes were similar in both groups. There was a two to threefold greater risk of liver cirrhosis and decompensated liver disease in the alcohol group. Also, the rate to which subjects developed cirrhosis was faster in the alcohol group with 58% being cirrhotic by the second decade as opposed to 10% being cirrhotic in the nonalcohol group by the second decade. The histologic and clinical acceleration of liver disease was independent of the mode of exposure or sex. In summary, alcohol intake is an independent risk factor in the clinical and histologic progression of HCV infection. PMID- 9731577 TI - Dissection of human humoral immune response against hepatitis C virus E2 glycoprotein by repertoire cloning and generation of recombinant Fab fragments. AB - Demonstration of antibodies inhibiting key viral functions is the basis for the design of an effective vaccine. Dissection of the human antibody response by repertoire cloning may be a powerful means to address this issue. In this study, a panel of human monoclonal recombinant Fab fragments specific for hepatitis C virus (HCV) E2 envelope protein was generated. The selection procedure was designed to select for cross-genotype reactive antibodies. Sequences coding five different human recombinant Fabs specific for the HCV/E2 protein were cloned and characterized. The ability of the cloned antibody fragments to inhibit adhesion of recombinant envelope E2 protein to target cells was assayed. While affinity of the different antibody fragments appeared similar, activity in inhibiting E2 binding to target cells varied considerably from one Fab fragment to another. Two Fabs were not able to inhibit E2 binding at high concentration (40 microg/mL), while three other Fab clones were active in neutralizing 50% of the E2 binding at concentrations ranging from 3 to 0.35 microg/mL. PMID- 9731578 TI - Cellular immunity to hepatitis C virus core protein and the response to interferon in patients with chronic hepatitis C. AB - To investigate the involvement of T-cell response against hepatitis C virus (HCV) antigens in viral clearance after interferon therapy, we measured interleukin-2 (IL-2) production by peripheral mononuclear cells in response to HCV core in patients with chronic hepatitis C. In a cohort of 43 patients, we investigated the frequency of circulating core-specific T-helper (Th) cell precursors by the limiting-dilution assay, and in a second cohort of 60 patients, we analyzed the response to specific core epitopes using 52 synthetic 15-mer overlapping peptides. We observed that the frequency of core-specific Th cell precursors was significantly higher in patients with sustained biochemical and virological response (SR) after interferon (IFN) therapy (median, 1/55,736) than in untreated patients (1/274,023) or that in patients who remained viremic after completion of the treatment-nonresponders (NR) plus transient responders (TR) (1/1,909,972). Patients who failed to respond to IFN (NR) and those who relapsed after IFN discontinuation (TR) had a similarly low number of precursors. The number of core peptides recognized by SR, TR, NR, UT, and healthy controls was 8.2 +/- 1.5, 6.5 +/- 1.2, 2.0 +/- 0.5, 2.7 +/- 0.9, and 0.3 +/- 0.2, respectively. In SR, the intensity of the proliferative response to core peptides as estimated by the summation of stimulation indexes (sigmaSI) was significantly higher than in NR and than in UT, but not different from that of TR. Our results indicate that both expansion of HCV-specific Th cell precursors and Th cell recognition of multiple core epitopes seem to be important in the elimination of HCV after IFN therapy. PMID- 9731579 TI - Predictors of patient and graft survival following liver transplantation for hepatitis C. AB - End-stage liver disease secondary to hepatitis C virus (HCV) infection is the leading indication for liver transplantation in the United States. Recurrence of HCV infection is nearly universal. We studied the patients enrolled in the National Institute of Diabetes and Digestive and Kidney Diseases Liver Transplantation Database to determine whether pretransplantation patient or donor variables could identify a subset of HCV-infected recipients with poor patient survival. Between April 15, 1990, and June 30, 1994, 166 HCV-infected and 509 HCV negative patients underwent liver transplantation at the participating institutions. Median follow-up was 5.0 years for HCV-infected and 5.2 years for HCV-negative recipients. Pretransplantation donor and recipient characteristics, and patient and graft survival, were prospectively collected and compared. Cumulative patient survival for HCV-infected recipients was similar to that of recipients transplanted for chronic non-B-C hepatitis, or alcoholic and metabolic liver disease, better than that of patients transplanted for malignancy or hepatitis B (P = .02 and P = .003, respectively), and significantly worse than that of patients transplanted for cholestatic liver disease (P = .001). Recipients who had a pretransplantation HCV-RNA titer of > or = 1 x 10(6) vEq/mL had a cumulative 5-year survival of 57% versus 84% for those with HCV-RNA titers of < 1 x 10(6) vEq/mL (P = .0001). Patient and graft survival did not vary with recipient gender, HCV genotype, or induction immunosuppression regimen among the HCV-infected recipients. While long-term patient and graft survival following liver transplantation for end-stage liver disease secondary to HCV are generally comparable with that of most other indications, higher pretransplantation HCV-RNA titers are strongly associated with poor survival among HCV-infected recipients. PMID- 9731580 TI - The efficacy of prophylactic interferon alfa-2b in preventing recurrent hepatitis C after liver transplantation. AB - Clinical recurrence of hepatitis C after liver transplantation can lead to cirrhosis, liver failure, and death. In patients undergoing liver transplantation for hepatitis C, we assessed the efficacy of interferon alfa-2b (IFN) in preventing recurrent hepatitis. We randomized 86 patients to either an IFN group (3 MU three times a week starting within 2 weeks after transplantation and continued for 1 year) or a control (no IFN) group. Recurrence, the primary end point, was diagnosed on biopsy performed at 1 year or for abnormal biochemistries. HCV RNA levels were measured by branched-chain DNA (bcDNA) assay and arbitrarily defined as low, moderate, or high (< 10 x 10(5), 10-100 x 10(5), or > 100 x 10(5) Eq/mL, respectively). Data on 30 IFN patients and 41 no-IFN patients who survived > or = 3 months were reviewed. Mean follow-up was 669 +/- 228 days for IFN patients and 594 +/- 254 days for no-IFN patients. IFN patients were less likely to develop recurrent hepatitis (8 IFN vs. 22 no-IFN patients, P = .017, log rank analysis). IFN and 1-month HCV RNA level were independent predictors of recurrence. IFN reduced the risk of recurrence by a factor of 0.4 (P = .04, Cox proportional hazards model); HCV RNA level > 100 x 10(5) Eq/mL at 1 month after transplantation increased the risk by a factor of 3.1 (P = .01). Low, moderate, and high viral levels at 1 and 3 months were associated with significantly different rates of recurrence in IFN patients (P = .05 at 1 month and P = .003 at 3 months) but not in untreated patients (P = .28 at 1 month and P = .25 at 3 months). In patients with two or more rejections, the risk of recurrence was increased by a factor of 2.17 (P = .05). On 47 1-year biopsies (24 IFN; 23 no IFN), piecemeal necrosis was more common in untreated patients (P < .02). One- and 2-year patient survival, respectively, was 96% and 96% with IFN and 91.2% and 87.2% without (P = NS). Prophylactic IFN reduced the incidence of recurrent hepatitis after transplant. Although IFN was most effective in patients with low HCV RNA levels, we also noted an effect in patients with moderate levels. IFN did not prevent viremia, suggesting that it may work through alternative mechanisms. PMID- 9731581 TI - TT-virus infection in North American blood donors, patients with fulminant hepatic failure, and cryptogenic cirrhosis. AB - A novel DNA virus, TT-virus (TTV), has been reported in patients with non-A-G posttransfusion hepatitis in Japan. We sought to determine whether TTV infection occurs in North American blood donors and to further determine the prevalence of TTV infection in several groups of patients with liver disease, including patients with cryptogenic cirrhosis and idiopathic fulminant hepatic failure. TTV infection was sought by detection of TTV DNA in serum by polymerase chain reaction (PCR) using primers generated from a conserved region of the TTV genome. Blood donors, patients with cryptogenic cirrhosis, idiopathic fulminant hepatic failure, and patients with other forms of advanced liver disease with and without a history of parenteral exposures were studied. TTV infection was present in 1% (1 of 100) of blood donors, 15% (5 of 33) of patients with cryptogenic cirrhosis, 27% (3 of 11) of patients with idiopathic fulminant hepatic failure, 18% (2 of 11) of patients with a history of exposure to blood products, and 4% (1 of 25) of patients without parenteral risk factors. For all patients tested, a history of prior exposure to blood products was associated with an increased risk of TTV infection (relative risk, 4.5; 90% confidence intervals, 0.6-43.9). We conclude that TTV infection is present among North American blood donors and is common in patients with liver disease, including cryptogenic cirrhosis and fulminant hepatic failure. Further studies are required to determine the role of TTV in the pathogenicity of acute and/or chronic liver disease. PMID- 9731582 TI - Rho directs activation-associated changes in rat hepatic stellate cell morphology via regulation of the actin cytoskeleton. AB - Hepatic stellate cell activation, thought to play a key role in fibrosis of the liver, is characterized by changes in cellular morphology. The intracellular signals regulating morphological alterations associated with stellate cell activation are uncertain. The ras-like guanosine triphosphate-binding protein, rho, has recently emerged as an important regulator of the actin cytoskeleton, and consequently cell morphology. The aim of this study was to test the hypothesis that rho signaling pathways direct activation-associated morphological changes in stellate cells by regulating the actin cytoskeleton. The morphology and actin cytoskeleton of primary rat hepatic stellate cells were studied with phase contrast, differential interference contrast, and epifluorescence microscopy. Immunohistochemistry and immunoblot analysis were used to examine rho expression and activity, respectively. Quiescent and activated stellate cells were investigated in the absence and presence of C3 transferase, a bacterial toxin that specifically inhibits rho. Stellate cell activation was characterized by the development of prominent intracellular fibers, and the loss of dendrite like processes and perinuclear retinoid droplets. Moreover, activation was accompanied by the formation of prominent actin stress fibers and focal adhesions. Both rho expression and activity were demonstrated in stellate cells. C3 transferase blocked and reversed, both activation-associated morphological alterations and activation-associated changes in the actin cytoskeleton, in quiescent and activated stellate cells, respectively. These results indicate that rho directs activation-associated changes in rat hepatic stellate cell morphology via regulation of the actin cytoskeleton. PMID- 9731583 TI - Hyponatremia in cirrhosis: from pathogenesis to treatment. PMID- 9731584 TI - Apoptosis: silent killer or neutron bomb? PMID- 9731586 TI - The Nineteenth United States-Japan Joint Hepatitis Panel Meeting. PMID- 9731585 TI - Portal hypertension and variceal bleeding: an AASLD single topic symposium. PMID- 9731587 TI - What the Amish can tell us about...cholestasis. PMID- 9731588 TI - Hereditary hemochromatosis--sometimes having a real complex can be a good thing. PMID- 9731589 TI - Chronobiological analysis of seasonal variations of variceal hemorrhage. PMID- 9731590 TI - Antiphospholipid-protein antibodies and ischemic stroke: not just cardiolipin any more. PMID- 9731591 TI - High prevalence of antiphosphatidylinositol antibodies in young patients with cerebral ischemia of undetermined cause. AB - BACKGROUND AND PURPOSE: Anticardiolipin antibodies (aCL) are associated with thrombotic phenomena including cerebral ischemia in young adults. Although aCL are directed to a neoepitope formed by phospholipid and beta2-glycoprotein I (beta2-GPI), immunoassays based on cardiolipin as target antigen are widely used. We previously demonstrated that 47% of aCL-negative systemic lupus erythematosus (SLE) patients had antiphospholipid antibodies (aPL) to epitopes other than cardiolipin, and we found an association between aPL to noncardiolipin antigens and thrombosis. We now assess the prevalence and clinical significance of noncardiolipin aPL in young adults with cerebrovascular disease of undetermined etiology. METHODS: Seventy-seven non-SLE patients, aged <51 years, with cerebral ischemia were studied. Specificity of aPL were characterized by ELISAs using 7 different phospholipids: cardiolipin (CL), phosphatidylserine (PS), phosphatidylinositol (PI), phosphatidylglycerol (PG), phosphatidic acid (PA), phosphatidylcholine, and phosphatidylethanolamine. RESULTS: Thirty-four patients (44.1%), had aPL to 1 or more of the following antigens: 23.4% to CL, 18.2% to PS, 15.6% to PG, 14.3% to PA, and 28.6% to PI. Fifty-nine patients (76.6%) were aCL negative. Of these subjects 23.4% showed aPL to noncardiolipin epitopes. PI was the specificity with highest prevalence in all subgroups, and in 6 patients anti-PI antibodies were the only detectable aPL. The binding of aPL to the different antigens was beta2-GPI dependent. CONCLUSIONS: Our data demonstrate a high prevalence of aPL in young adults with cerebral ischemia of undetermined cause. PI was the specificity with highest prevalence, suggesting that anti-PI antibodies may be an immunological marker in young patients with cerebrovascular disease. PMID- 9731592 TI - Prothrombin gene G20210-->A transition is a risk factor for cerebral venous thrombosis. AB - BACKGROUND AND PURPOSE: It has been recently reported that a G-->A transition at nucleotide position 20210 in the 3'-untranslated region of the prothrombin gene is associated with elevated plasma prothrombin levels and an increased risk of deep venous thrombosis. To date, it is unknown whether this polymorphism also represents a risk factor for cerebral venous thrombosis (CVT). METHODS: Venous blood samples were collected from 45 patients with CVT and from 354 healthy blood donors as controls. A second control group consisted of 131 subjects with acute ischemic stroke or transient ischemic attack (TIA). Genomic DNA was isolated from peripheral blood leukocytes. Amplification of DNA was performed by polymerase chain reaction (PCR). The G-->A transition at nucleotide position 20210 of the prothrombin gene was detected by allele-specific restriction digestion. RESULTS: The G20210-->A transition in the prothrombin gene was found in a heterozygous form in 4 of 45 patients with CVT (8.9%) and in 8 of 354 healthy control subjects (2.3%). This difference was statistically significant (P=0.010). The G20210-->A transition increased the relative risk for CVT approximately 5-fold (age-adjusted odds ratio 5.7; 95% CI 1.5 to 21.5). In contrast, in the group of patients with acute cerebral ischemia, only 3 of 131 subjects (2.3%) were heterozygous for the G20210-->A transition, which corresponded to the prevalence in the group of healthy blood donors. CONCLUSIONS: The recently described G20210-->A transition in the 3'-untranslated region of the prothrombin gene is an inherited risk factor for CVT but obviously not for acute ischemic stroke or TIA. PMID- 9731593 TI - Mechanisms of cerebrovascular events as assessed by procoagulant activity, cerebral microemboli, and platelet microparticles in patients with prosthetic heart valves. AB - BACKGROUND AND PURPOSE: Cerebrovascular events (CVE) in patients with prosthetic heart valves (PHV) have remained a severe and frequent complication despite oral anticoagulation with or without aspirin. We studied the possible pathophysiological involvement of platelet-derived microparticles (PMP) as a contributing factor for the increased incidence of CVE in patients with PHV. METHODS: We compared in a retrospective, case-control study the clinical outcome after the implantation of the PHV with several different independent morphological and functional methods, including simultaneous transcranial Doppler monitoring of both middle cerebral arteries, PMP detection by flow cytometry with use of platelet-specific antibodies, coagulation markers, and determination of the procoagulant activity by Russell's viper venom time, a phospholipid-dependent coagulation assay. RESULTS: Eight of 26 patients with PHV had 9 CVE during 136 person-years of observation. Transcranial Doppler monitoring revealed an increased frequency of microembolic signals recorded over a 30-minute period in patients with CVE (75+/-25; median, 55; range, 27 to 248) compared with those without CVE (23+/-12; median, 7; range, 0 to 153; P<0.05) or with control subjects (0; P<0.001). Flow cytometry analysis showed increased levels of PMP in patients with compared to those without CVE (4.1+/-0.6% versus 2.4+/-0.4% of all fluorescence-positive events gated; P<0.05). Increased procoagulant activity was documented by the shortened Russell's viper venom time expressed as an increased level of platelet equivalents per microliter of plasma in patients compared with control subjects (+24.7+/-14.9%; P<0.01). Subgroup analysis revealed that patients with CVE had a higher excess of platelet equivalents per microliter of plasma than patients without CVE in relation to the controls (+68.7+/-36.7%; P<0.05). Mildly elevated thrombin-antithrombin III complexes (2.9+/-0.7; median, 2.3; normal, <2.0 microg/L) suggested incompletely suppressed thrombin formation, and fibrin generation (fibrinopeptide A) was in the upper normal range (2.1+/ 0.2; median, 1.8; normal, <2.0 ng/mL), despite adequate anticoagulation (INR=3.6+/-0.1). CONCLUSIONS: Our data show increased microembolic signals, platelet microparticles, and procoagulant activity in symptomatic patients with PHV and provide a potential pathophysiological explanation of CVE. PMID- 9731594 TI - Time course of ADCw changes in ischemic stroke: beyond the human eye! AB - BACKGROUND AND PURPOSE: Using newly developed computerized image analysis, we studied the heterogeneity of apparent diffusion coefficient of water (ADCw) values in human ischemic stroke within 10 hours of onset. METHODS: Echo-planar trace diffusion-weighted images from 9 patients with focal cortical ischemic stroke were obtained within 10 hours of symptom onset. An Iterative Self Organizing Data Analysis (ISODATA) clustering algorithm was implemented to segment different tissue types with a series of DW images. ADCw maps were calculated from 4 DW images on a pixel-by-pixel basis. The segmented zones within the lesion were characterized as low, pseudonormal, or high, expressed as a ratio of the mean+/-SD of ADCw of contralateral noninvolved tissue. RESULTS: The average ADCW in the ischemic stroke region within 10 hours of onset was significantly depressed compared with homologous contralateral tissue (626.6+/ 76.8 versus 842.9+/-60.4x10(-6) mm2/s; P<0.0001). Nevertheless, ISODATA segmentation yielded multiple zones within the stroke region that were characterized as low, pseudonormal, and high. The mean proportion of low:pseudonormal:high was 72%:20%:8%. CONCLUSIONS: Despite low average ADCW, computer-assisted segmentation of DW MRI detected heterogeneous zones within ischemic lesions corresponding to low, pseudonormal, and high ADCw not visible to the human eye. This supports acute elevation of ADCw in human ischemic stroke and, accordingly, different temporal rates of tissue evolution toward infarction. PMID- 9731595 TI - Diffusion-weighted magnetic resonance imaging in acute stroke. AB - BACKGROUND AND PURPOSE: Diffusion-weighted MRI (DWI) is highly sensitive in detecting early cerebral ischemic changes in acute stroke patients. In this study we compared the sensitivity of DWI with that of conventional MRI techniques. Furthermore, we investigated the prognostic value of the volume of ischemic lesions on DWI scans and of the apparent diffusion coefficient (ADC). METHODS: We performed DWI, fluid-attenuated inversion recovery, spin-echo T2-weighted MRI, and spin-echo proton density-weighted MRI in 42 patients with acute stroke and 15 control subjects. The volume of ischemic lesions was measured on early (<60 hours after onset) and follow-up MRI scans. Clinical outcome was measured 4 months after onset of symptoms with the National Institutes of Health Stroke Scale, the Barthel Index, and the Rankin Scale. RESULTS: With DWI, 98% of the ischemic lesions were detected, and with fluid-attenuated inversion recovery, 91% were detected, whereas with early T2-weighted or proton density-weighted scans, only 71% (P=0.002, chi2) and 80% (P=0.02, chi2) of lesions, respectively, were found. Lesion volume on early DWI scans correlated significantly with clinical outcome ratings (P<0.01). In patients with a first-ever stroke, a lesion volume of < or =22 mL on DWI predicted good outcome with a 75% sensitivity and a 100% specificity. The mean ADC of ischemic lesions was 29% lower than the ADC of normal-appearing parts of the brain (P<0.001). The ADC ratio correlated significantly with clinical outcome (P<0.05). CONCLUSIONS: DWI is a better imaging method than conventional MRI in detecting early ischemic lesions in stroke patients. Lesion size as measured on DWI scans and, to a lesser extent, ADC values are potential parameters for predicting clinical outcome in acute stroke patients. PMID- 9731596 TI - Perilesional blood flow and edema formation in acute intracerebral hemorrhage: a SPECT study. AB - BACKGROUND AND PURPOSE: Secondary brain injury and edema formation contribute significantly to morbidity and mortality after intracerebral hemorrhage (ICH). The pathogenesis of this process is poorly understood. We sought to characterize alterations in perilesional blood flow that occur during the acute phase of ICH and to determine whether progressive enlargement of edema surrounding ICH is related to increased or decreased perfusion. METHODS: We performed paired consecutive CT and 99mTc-hexamethylpropylenamine oxime single-photon emission computed tomography (SPECT) scans during the acute (mean, 18 hours) and subacute (mean, 72 hours) phase of ICH in 23 patients. Hematoma and edema volumes were traced and calculated from CT images. SPECT-derived hypothetical flow deficit volumes (FDV) around each hematoma were calculated by measuring a "zero-flow" volume within a large perilesional region of interest (based on percent tracer count loss compared with the contralateral side) and subtracting the corresponding ICH volume. Patients with significant midline shift (>5 mm) or global blood flow reduction were excluded from the analysis. RESULTS: ICH volume (18 mL) did not change, mean edema volume increased by 36% (from 19 to 25 mL, P<0.0001), and mean FDV decreased by 55% (from 14 to 6 mL, P=0.0004) between the acute and subacute phases. Edema volume on the second CT scan correlated positively with FDV on the first SPECT scan (Spearman's p=0.48, P=0.02), and with the volume of reperfused perilesional tissue (FDVacute-FDVsubacute) (Spearman's p=0.41, P=0.05). Perilesional edema on CT always corresponded topographically with perfusion deficits on SPECT. In 4 patients, delayed focal hyperemia was identified in more peripheral cortical regions, but these areas appeared normal on CT. CONCLUSIONS: Perilesional blood flow normalizes from initially depressed levels as edema forms during the first 72 hours after ICH, and the eventual extent of edema correlates with the volume of reperfused tissue. These results suggest that the potential for perilesional ischemia is highest in the earliest hours after ICH onset and implicate reperfusion injury in the pathogenesis of perihematoma edema formation. PMID- 9731597 TI - Comparison of the ABC/2 estimation technique to computer-assisted volumetric analysis of intraparenchymal and subdural hematomas complicating the GUSTO-1 trial. AB - BACKGROUND AND PURPOSE: The volume of an intracerebral hemorrhage has been shown to be an important independent predictor of mortality in several reports. A technique for estimating hematoma volume, known as the ABC/2 method, has been proven a reliable, simple bedside technique for the volume measurement of intraparenchymal intracerebral hemorrhage. Subdural hematomas also carry a significant mortality risk but are more amenable to surgical evacuation. A reliable, simple bedside measurement of subdural hematoma volume may prove a valuable tool in prognostication and management of patients with this entity. METHODS: Computed tomographic (CT) brain scans of 244 patients suffering from intracranial hemorrhage in the GUSTO-1 trial were systematically reviewed. The volumes of 298 intraparenchymal hematomas were measured by the ABC/2 technique, and the volumes of 44 subdural hematomas were measured by an adaptation of this technique and compared to computer-assisted volumetric analysis. RESULTS: Excellent correlation between the techniques were achieved for both subdural (r=0.842; slope, 0.982) and intraparenchymal hematoma volume measurements (r=0.929; slope, 1.11). CONCLUSIONS: The ABC/2 method is a simple and accurate technique for the measurement of intraparenchymal hematoma volume, and a simple adaptation allows for a similarly accurate measurement of subdural hematoma volume as well. PMID- 9731598 TI - Recurrent primary cerebral hemorrhage: frequency, mechanisms, and prognosis. AB - BACKGROUND AND PURPOSE: The frequency of recurrent primary cerebral hemorrhage (RPCH), mainly in cases related to hypertension, has been considered low. This study investigated the frequency, mechanisms, and prognosis of RPCH. METHODS: We evaluated 359 patients with neuroimaging evidence of cerebral hemorrhage and selected 22 with RPCH. RESULTS: Five patients (23%) were older than 70 years at the first cerebral hemorrhage. Mean ages at the first and second hemorrhages were 60 and 63 years, respectively. Risk factors included hypertension (86%), diabetes (27%), and tobacco and alcohol use (each 14%). Hypocholesterolemia was demonstrated in 35% of the patients. The most common pattern of recurrent bleeding was ganglionic-ganglionic, mainly related to hypertension. Overall mortality was 32%. Forty-one percent and 27% of patients, respectively, had incapacitating and nonincapacitating sequelae; 2 of the latter had RPCH with a lobar location. Ganglionic-ganglionic hemorrhage was associated with a poor prognosis; otherwise, this pattern was uncommon in patients with nonincapacitating sequelae. Analysis of the control of risk factors, primarily hypertension after the first cerebral hemorrhage, disclosed that 56% of patients did not gain subsequent control. CONCLUSIONS: Rebleeding after a first primary intracerebral hemorrhage is not uncommon. The main topographic pattern of bleeding, ganglionic-ganglionic, is likely the result of hypertension; the less common lobar-lobar pattern probably results from amyloid angiopathy. PMID- 9731599 TI - Blood pressure control and recurrence of hypertensive brain hemorrhage. AB - BACKGROUND AND PURPOSE: Recent studies have demonstrated that recurrence of hypertensive brain hemorrhage (HBH) is not uncommon. However, risk factors for the recurrence of HBH have not been evaluated systematically. METHODS: We analyzed 74 patients with HBH who were admitted to our clinic and followed up as outpatients for a mean of 2.8 years. Blood pressure (BP) and other clinical features were compared between the groups of patients with and without rebleeding. We determined the recurrence rate of HBH in relation to BP. RESULTS: Diastolic BP was significantly higher in the recurrence group than in the nonrecurrence group (88+/-8 versus 82+/-7 mm Hg; P=0.04). Systolic BP and other clinical variables were not different between the groups. The recurrence rate was 10.0% per patient-year in patients with diastolic BP >90 mm Hg and <1.5% in those with lower diastolic BP (P<0.001). No patients with diastolic BP <70 mm Hg experienced rebleeding. CONCLUSIONS: Higher diastolic BP was related to an increased rate of rebleeding. Diastolic BP >90 mm Hg may be regarded as a factor predictive of the recurrence of HBH. PMID- 9731600 TI - Asymptomatic embolization in subjects with atrial fibrillation not taking anticoagulants: a prospective study. AB - BACKGROUND AND PURPOSE: Embolism is believed to be the major cause of stroke in patients with nonvalvular atrial fibrillation (NVAF). The detection of asymptomatic embolic signals (ES) in individuals with NVAF might allow identification of patients at high risk of stroke and monitoring of therapy in individual subjects. We determined the frequency of asymptomatic ES in patients with NVAF who were not taking warfarin. METHODS: Bilateral transcranial Doppler recordings were made for 1 hour from the middle cerebral arteries of 111 successive patients with NVAF taking aspirin alone or no antithrombotic or anticoagulant therapy. Adequate recordings could be made in 86 patients. In 79 subjects, recordings were performed on a second occasion to study temporal variability. Recordings for a single hour were also made in 30 age-matched control subjects. RESULTS: ES were detected in 13 (15.1%) of NVAF subjects but in no control subjects (P=0.02). ES were detected both in subjects with symptomatic NVAF (4 of 30 [13.1%], P=0.04 versus controls) and asymptomatic NVAF (9 of 56 [16.1%], P=0.02 versus controls). There was no correlation between the presence of ES and smoking status, diabetes, hypertension, aspirin use, aspirin dose, symptomatic status, left atrial size, left ventricular function, or the presence of left atrial thrombus detected on transthoracic echocardiography. Repeating the recording increased the number of patients with ES to 21 (26.6%). On considering the results of both recordings, again there was no association for either recording between the presence of ES and smoking status, diabetes, hypertension, aspirin use, aspirin dose, age, symptomatic status, left atrial size, or left ventricular function. On repeating the recording, in the symptomatic group only 2 patients (8%) changed status, in contrast to 15 (29%) in the asymptomatic group. CONCLUSIONS: ES can be detected in patients with NVAF at a low frequency. Particularly in asymptomatic patients, ES show marked temporal variability. We found no correlation between the presence of previously reported clinical and echocardiographic markers of increased stroke risk and the presence of ES. This association requires further investigation before the clinical utility of this technique in patients with NVAF is decided. PMID- 9731601 TI - Atrial fibrillation is an independent determinant of low cognitive function: a cross-sectional study in elderly men. AB - BACKGROUND AND PURPOSE: Cerebrovascular disease is increasingly recognized as a cause of dementia and cognitive decline. We have previously reported an association between hypertension and diabetes and low cognitive function in the elderly. Atrial fibrillation is another main risk factor for cerebrovascular disease. The aim of this study was to investigate whether atrial fibrillation is associated with low cognitive function in elderly men with and without previous manifest stroke. METHODS: This was a cross-sectional study based on a cohort of 952 community-living men, aged 69 to 75 years, in Uppsala, Sweden. Cognitive functions were assessed by the Mini-Mental State Examination and the Trail Making Tests, and a composite z score was calculated. The relation between atrial fibrillation and cognitive z score was analyzed, with stroke and other vascular risk factors taken into account. RESULTS: All analyses were adjusted for age, education, and occupational level. Men with atrial fibrillation (n=44) had lower mean adjusted cognitive z scores (-0.26+/-0.11) than men without atrial fibrillation (+0.14+/-0.03; P=0.0003). The exclusion of stroke patients did not alter this relationship; the mean cognitive z score was -0.24+/-0.12 in the 36 men with atrial fibrillation and +0.17+/-0.03 in those without atrial fibrillation (P=0.0004), corresponding to a difference of 0.4 SDs between groups. Adjustments for 24-hour diastolic blood pressure and heart rate, diabetes, and ejection fraction did not change this relationship. Men with atrial fibrillation who were treated with digoxin (n=27) performed markedly better (-0.05+/-0.21) than those without treatment (n=9; -1.14+/-0.34; adjusted P=0.0005). Previous myocardial infarction was not associated with impaired cognitive results. CONCLUSIONS: In these community-living elderly men, we found an association between atrial fibrillation and low cognitive function independent of stroke, high blood pressure, and diabetes. Interventional studies are needed to answer the question of whether optimal treatment of atrial fibrillation may prevent or postpone cognitive decline and dementia. PMID- 9731602 TI - Microemboli in cerebral circulation and alteration of cognitive abilities in patients with mechanical prosthetic heart valves. AB - BACKGROUND AND PURPOSE: It has been shown previously that cerebral microemboli may occur frequently in patients with a normal mechanical heart valve (MHV) without prior history of stroke. Some arguments strongly suggest that these microemboli have a gaseous origin. In other circumstances such as extracorporeal circulation or decompression in divers, it has been demonstrated that cerebral microbubbles could lead to some deterioration in cognitive functions. Therefore, we have studied attention and memory, which are among the most impaired cognitive functions as demonstrated in previous studies, in patients with an MHV. METHODS: Three groups of 12 volunteers each were composed of patients with an MHV and embolic signals in the cerebral circulation (group 1), patients with biological prostheses (group 2), and healthy subjects (group 3). Groups were carefully matched for age and verbal intellectual abilities. For each group, a transcranial Doppler examination was performed and a set of cognitive tests assessing sustained and selective attention and episodic and working memory was administered. RESULTS: The mean embolic rate was 29 per hour in patients with an MHV. No embolus was detected in the other 2 groups. Episodic memory was significantly modified in both groups 1 and 2 compared with the control group for tasks that required high-processing resources. Working memory performance was significantly decreased in MHV patients. No between-groups differences were observed for the other parameters. CONCLUSIONS: Alteration of episodic memory can be attributed to a long-term effect of the surgical procedure. Deterioration of working memory can be related to the presence of cerebral microemboli in MHV patients. PMID- 9731603 TI - Cost-effectiveness of anticoagulation in nonrheumatic atrial fibrillation in the primary prevention of ischemic stroke. AB - BACKGROUND AND PURPOSE: A number of clinical trials have shown the value of anticoagulating patients with nonrheumatic atrial fibrillation to prevent ischemic stroke. The purpose of this study was to assess the cost-effectiveness of anticoagulation in nonrheumatic atrial fibrillation with particular reference to the very elderly (aged >75 years) who have a higher incidence of bleeding events while undergoing anticoagulation. METHODS: We calculated the incremental costs per life-year gained for 4 base cases using efficacy data from the Boston Area Anticoagulation Trial for Atrial Fibrillation, the meta-analysis of the 5 nonrheumatic atrial fibrillation trials, cost data from a district general hospital, and review of the literature. RESULTS: The cost per life-year gained free from stroke over 10 years ranged from -pound sterling 400.45 (ie, a resource saving achieved for each life-year gained free from stroke) to pound sterling 13,221.29. The results were most sensitive to alteration in the frequency of anticoagulation monitoring. CONCLUSIONS: For medical and economic reasons, anticoagulation treatment in the prevention of ischemic stroke is justified. Although older patients are more at risk of adverse events, anticoagulation is more cost-effective in this group. PMID- 9731605 TI - Prediction of functional outcome after stroke: comparison of the Orpington Prognostic Scale and the NIH Stroke Scale. AB - BACKGROUND AND PURPOSE: This study compared the ability of 2 stroke impairment scales, Orpington Prognostic Scale and National Institutes of Health (NIH) Stroke Scale, to predict disability as measured by the Barthel activities of daily living (ADL) Index and higher level of self-reported physical functioning as measured by the SF-36 physical functioning index (PFI) at 1, 3, and 6 months after stroke. METHODS: The participants in this ongoing study are 184 individuals who sustained an eligible stroke and were recruited for the Kansas City Stroke Study. All patients were prospectively evaluated using standardized assessments at enrollment (within 14 days of stroke onset) and followed at 1, 3, and 6 months after stroke. Coefficient of determination (R2) was used to assess the ability of the 2 stroke scales to prognosticate outcomes. RESULTS: Means and SDs of the Orpington Prognostic Scale and NIH Stroke Scale measured at baseline were 3.6+/ 1.31 and 5.5+/-4.58, respectively. The Spearman's rank correlation between the 2 baseline measures was 0.83 (P=0.0001). The Orpington Prognostic Scale and the NIH Stroke Scale explained well the variance in Barthel ADL Index (P<0.001). However, the Orpington Prognostic Scale explained more variance than did the NIH Stroke Scale. Similarly, the Orpington Prognostic Score explained more variance in higher level of physical function than did the NIH Stroke Scale. The amount of variance in Barthel ADL Index and SF-36 PFI, which were explained by both stroke severity measures, decreased over time. CONCLUSIONS: Our results demonstrate that in a sample of mostly mild and moderate strokes, the Orpington Prognostic Scale compared with the NIH Stroke Scale is simpler to use and is a slightly better predictor of ADL and higher levels of physical function. PMID- 9731604 TI - Association between mitral annulus calcification and carotid atherosclerotic disease. AB - BACKGROUND AND PURPOSE: It has been established that mitral annulus calcification (MAC) is an independent predictor of stroke, though a causative relationship was not proved, and that carotid artery atherosclerotic disease is also associated with stroke. The aim of this study was to determine whether there is an association between the presence of MAC and carotid artery atherosclerotic disease. METHODS: Of the 805 patients in whom the diagnosis of MAC was made by transthoracic echocardiography between 1995 and 1997, 133 patients (60 men and 73 women; mean age, 74.3+/-8 years; range, 47 to 89 years) underwent carotid artery duplex ultrasound for various indications; the study group comprised these patients. They were compared with 129 age- and sex-matched patients without MAC (57 men and 72 women; mean age, 73.6+/-7 years; range, 61 to 96 years) who underwent carotid artery duplex ultrasound during the same period for the same indications. MAC was defined as a dense, localized, highly reflective area at the base of the posterior mitral leaflet. MAC was considered severe when the thickness of the localized, highly reflective area was > or =5 mm on 2 dimensional echocardiography in the 4-chamber view. Carotid artery stenosis was graded as follows: 0%, 20%, 40%, 60%, 80%, and 100%. RESULTS: Compared with the control group, the MAC group showed a significantly higher prevalence of carotid stenosis of > or =40% (45% versus 29%, P=0.006), which was associated with > or =2-vessel disease (23% versus 10%, P=0.006) and bilateral carotid artery atherosclerotic disease (21% versus 10%, P=0.011). Severe MAC was found in 48 patients. More significant differences were found for the severe MAC subgroup (for carotid stenosis of > or =40%) in rates of carotid artery atherosclerotic disease (58% versus 29%, P=0.001), and > or =2-vessel disease (31% versus 10%, P=0.001), in addition to bilateral carotid artery stenosis (27% versus 10%, P=0.004) and even bilateral proximal internal carotid artery stenosis (21% versus 8%, P=0.015). Furthermore, significant carotid artery atherosclerotic disease (stenosis of > or =60%) was significantly more common in the severe MAC subgroup than in the controls (42% versus 26%, P<0.05) and was associated with higher rates of > or =2-vessel disease (19% versus 7%, P=0.02) and bilateral carotid artery stenosis (17% versus 7%, P=0.05). On multivariate analysis, MAC and age but not traditional risk factors were the only independent predictors of carotid atherosclerotic disease (P=0.007 and P=0.04, respectively). CONCLUSIONS: There is a significant association between the presence of MAC and carotid artery atherosclerotic disease. MAC may be an important marker for atherosclerotic disease of the carotid arteries. This association may explain the high prevalence of stroke in patients with MAC. PMID- 9731606 TI - Dependence and perceived difficulty in daily activities in community-living stroke survivors 2 years after stroke: a study of instrumental structures. AB - BACKGROUND AND PURPOSE: There is a need for better understanding of the structure of instruments for functional outcome assessment after discharge from rehabilitation. One purpose of the study was to contribute to the analysis of instrumental dimensionality. Another purpose was to compare disability in stroke patients within the younger age range 2 years after onset of stroke with that at discharge with respect to both dependence and patients' perceived difficulty and to extend the assessments with instrumental activities. METHODS: We studied 68 stroke survivors aged 18 to 71 years at onset (59% aged <55 years) by means of interviews in their home, using activities from the Functional Independence Measure (FIM) and Instrumental Activity Measure (IAM) for ratings of dependence and perceived difficulty. Rasch analysis was used to construct calibrated linear measures and to evaluate the level of fit. RESULTS: Acceptable models for comparison of dependence between discharge and follow-up were found for the physical and the social-cognitive items in FIM. However, personal care and social cognitive items showed an increased level of dependence at follow-up compared with at discharge. A combination of physical activities from FIM and IAM also gave acceptable models for both dependence and perceived difficulty, and the hierarchical orders of activities are presented. In general, there was agreement between the ratings of dependence and perceived difficulty, but with some discrepancies. Men found it harder to be independent in such instrumental activities as cooking and cleaning than women; the opposite was true for small scale shopping and locomotion outdoors. Subjects aged > or =55 years had slightly higher level of dependence and perceived difficulty in IAM activities than those below that age. CONCLUSIONS: Changes in the hierarchical order of activities should be taken into account in follow-up studies. Differences in the environment between hospital and home, as well as differences in support and motivation, might explain the relatively larger degree of dependence at follow-up compared with at discharge and indicate the need for further rehabilitation efforts. Instrumental activities could be combined with FIM activities in a model. For individual items, ratings of both dependence and perceived difficulty may provide further insight into the disablement process. PMID- 9731607 TI - Blood pressure and functional recovery in acute ischemic stroke. AB - BACKGROUND AND PURPOSE: The relevance of elevated blood pressure in acute ischemic stroke and its most appropriate management are unresolved. We aimed to evaluate the rate of functional recovery with relation to early blood pressure management in patients with ischemic stroke. METHODS: Four hundred eighty-one consecutive ischemic stroke patients were admitted to the Neurology Service within 20.9+/-10.5 hours of symptoms onset as part of the Barcelona Downtown Stroke Registry, including 235 patients who received oral antihypertensive agents within <24 hours after stroke onset. Demographic, clinical (Mathew scale), and CT scan findings were collected prospectively. Mean arterial pressure (MAP) was recorded before hospital arrival and at 7 AM on days 1, 2, and 7 of hospitalization. The primary end point was complete functional recovery at day 7 defined as a score of 0 to 1 on the modified Rankin scale. RESULTS: Two hundred fifty-two patients achieved complete recovery on day 7. Using logistic regression, independent predictors of complete recovery included mild impairment at stroke presentation, lack of history of hypertension, and absence of brain edema on CT scan. Also, a 20% to 30% drop in MAP on day 2 after stroke onset almost tripled the odds of full recovery (odds ratio, 2.9; 95% CI, 1.3 to 6.3). MAP tended to normalize after stroke in all subjects, more rapidly if hypotensive agents were administered. Brain edema was also less frequent in patients with a greater drop in blood pressure. Despite the fact that a drop in MAP >30% from baseline was observed in 49 patients, this preceded worsening stroke in only 4 patients. Conversely, worsening stroke occurred in 51 patients despite stable blood pressure. CONCLUSIONS: These results suggest that complete recovery in ischemic stroke is facilitated by a moderate blood pressure reduction when brain edema develops, most likely as the result of a more adequate cerebral perfusion pressure. Conversely, stroke worsening due to pharmacological hypoperfusion is exceptional. PMID- 9731608 TI - Prognostic value of motor evoked potential obtained by transcranial magnetic brain stimulation in motor function recovery in patients with acute ischemic stroke. AB - BACKGROUND AND PURPOSE: The early prognostic application of transcranial magnetic brain stimulation (TMS) for assessing motor and functional recovery in ischemic stroke patients has yielded contradictory results. We performed a prospective study of patients with acute ischemic stroke and motor deficit to evaluate the early prognostic value of TMS in motor and functional recovery. METHODS: Fifty patients with different degrees of hemiparesis were studied in the first week after ischemic stroke and evaluated by clinical scales (Medical Research Council Scale, Canadian Neurological Scale, Barthel Index), with clinical follow-up over 6 months. TMS (Magstim 200) was performed at the same time, recording the motor evoked potential (MEP) in the thenar eminence muscles, with facilitation by voluntary contraction. RESULTS: Of the total group of 50 patients, MEP was absent in 20 and present in 30 (17 with normal and 13 with delayed central conduction time [CCT]). The patients with MEP showed better motor and functional recovery than those without. The MEP provided information on patient recovery, regardless of the initial strength and/or Barthel values. The degree of recovery was better in those patients with normal CCT than in those with delayed CCT. CONCLUSIONS: MEP obtained by TMS in patients with hemiparesis after acute ischemic stroke is useful as an early prognostic indicator of motor and functional recovery. This technique would allow the early identification of those patients who will have a good recovery, particularly among those with severe initial paresis. PMID- 9731609 TI - Hyperinsulinemia and the risk of stroke in healthy middle-aged men: the 22-year follow-up results of the Helsinki Policemen Study. AB - BACKGROUND AND PURPOSE: Several studies have shown that hyperinsulinemia is associated with the risk of coronary heart disease, but information on the association of hyperinsulinemia with the risk of stroke is limited. We investigated the association of hyperinsulinemia with the risk of stroke during a 22-year follow-up of the Helsinki Policemen Study population. METHODS: The study was based on a cohort of 970 men aged 34 to 64 years who were free of cerebrovascular disease, other cardiovascular disease, or diabetes. Risk factor measurements at baseline examination included an oral glucose tolerance test with blood glucose and plasma insulin measurements at 0, 1, and 2 hours. Area under the insulin response curve during oral glucose tolerance test was used as a composite variable reflecting plasma insulin levels. RESULTS: During the 22-year follow-up, 70 men had a fatal or nonfatal stroke. Hyperinsulinemia (highest area under the insulin response curve quintile compared with the combined 4 lower quintiles) was associated with the risk of stroke (age-adjusted hazard ratio, 2.12; 95% CI, 1.28 to 3.49), but not independently of other risk factors (multiple-adjusted hazard ratio, 1.54; 95% CI, 0.90 to 2.62), which was mainly due to the impact of obesity, particularly upper body obesity, with subscapular skinfold thickness used as an index. Of other risk factors, upper body obesity, blood pressure, and smoking were independent predictors of the risk of stroke. CONCLUSIONS: Hyperinsulinemia was associated with the risk of stroke in Helsinki policemen during the 22-year follow-up, but not independently of other risk factors, particularly upper body obesity. PMID- 9731610 TI - A quantitative study of the emotional outcome of people caring for stroke survivors. AB - BACKGROUND AND PURPOSE: Although the physical and, to a lesser extent, emotional outcome of stroke survivors has been well documented, there are far fewer data relating to the outcomes of those who care for them. We aimed to describe the outcome of those caring for stroke patients and to identify both patient and caregiver factors that are associated with poor caregiver outcomes. METHODS: As part of a randomized trial to evaluate a stroke family care worker, we identified 417 patients (67% of all referrals to our institution). We followed up 376 survivors of whom 246 identified a caregiver at a 6-month follow-up interview. The patients and caregivers were asked to complete 2 measures of emotional distress ( 30-item General Health Questionnaire [GHQ-30] and Hospital Anxiety and Depression [HAD] Scale). A regression analysis was used to identify factors that were independently associated with poor caregiver outcomes. RESULTS: Fifty-five percent of responding caregivers scored more than 4 on the GHQ-30, indicating that emotional distress is common in this group. Caregivers were more likely to be depressed if the patients were severely dependent (P<0.01) or emotionally distressed themselves (P<0.01). Female caregivers reported more anxiety (median HAD=8) than male caregivers (median HAD=5; P<0.01) but caregivers' levels of anxiety were not so clearly related to the patients' degree of physical disability as their levels of depression. Caregivers suffered more emotional distress if the patients had been dependent before their strokes. CONCLUSIONS: These data may help to identify those caregivers at greatest risk of poor emotional outcomes and thus help in the planning of trials and delivery of interventions aimed at preventing or treating distress among caregivers. PMID- 9731611 TI - Circadian variation in acute ischemic stroke: a hospital-based study. AB - BACKGROUND AND PURPOSE: We investigated circadian rhythm in ischemic stroke onset and its subtypes, differentiating between first-ever stroke and recurrent stroke. METHODS: A consecutive series of 1223 patients with ischemic stroke was admitted at 2 reference hospitals; the time of onset of symptoms was obtained, differentiating between onset while asleep and awake. We compared circadian rhythm between stroke types and between first-ever and recurrent stroke. RESULTS: The onset time was known in 914 patients; 25.6% experienced onset on awakening [higher incidence in thrombotic and lacunar stroke (28.9% and 28.4%, respectively) than in embolic stroke (18.8%)]. For all stroke subtypes, there was a significant diurnal variation, with a morning peak between 6 AM and noon; after redistributing the hour of onset of patients awakening with stroke, the morning peak was minimal in all types of stroke. There were no differences in circadian rhythm between patients with first-ever and recurrent stroke. CONCLUSIONS: Only hospitalized patients were studied. There is a circadian rhythm in all types of stroke, with higher frequency during the day and lower frequency in the last hours in the evening. The highest incidence in the early hours of the morning can be overestimated, due to patients who awaken with stroke. There is no difference in circadian rhythm between first-ever stroke and recurrent stroke. PMID- 9731612 TI - Hypocapnia and cerebral hypoperfusion in orthostatic intolerance. AB - BACKGROUND AND PURPOSE: Orthostatic and other stresses trigger tachycardia associated with symptoms of tremulousness, shortness of breath, dizziness, blurred vision, and, often, syncope. It has been suggested that paradoxical cerebral vasoconstriction during head-up tilt might be present in patients with orthostatic intolerance. We chose to study middle cerebral artery (MCA) blood flow velocity (BFV) and cerebral vasoregulation during tilt in patients with orthostatic intolerance (OI). METHODS: Beat-to-beat BFV from the MCA, heart rate, CO2, blood pressure (BP), and respiration were measured in 30 patients with OI (25 women and 5 men; age range, 21 to 44 years; mean age, 31.3+/-1.2 years) and 17 control subjects (13 women and 4 men; age range, 20 to 41 years; mean age, 30+/-1.6 years); ages were not statistically different. These indices were monitored during supine rest and head-up tilt (HUT). We compared spontaneous breathing and hyperventilation and evaluated the effect of CO2 rebreathing in these 2 positions. RESULTS: The OI group had higher supine heart rates (P<0.001) and cardiac outputs (P<0.01) than the control group. In response to HUT, OI patients underwent a greater heart rate increment (P<0.001) and greater reductions in pulse pressure (P<0.01) and CO2 (P<0.001), but total systemic resistance failed to show an increment. Among the cerebrovascular indices, all BFVs (systolic, diastolic, and mean) decreased significantly more, and cerebrovascular resistance (CVR) was increased in OI patients (P<0.01) compared with control subjects. In both groups, hyperventilation induced mild tachycardia (P<0.001), a significant reduction of BFV, and a significant increase of CVR associated with a fall in CO2. Hyperventilation during HUT reproduced hypocapnia, BFV reduction, and tachycardia and worsened symptoms of OI; these symptoms and indices were improved within 2 minutes of CO2 rebreathing. The relationships between CO2 and BFV and heart rate were well described by linear regressions, and the slope was not different between control subjects and patients with OI. CONCLUSIONS: Cerebral vasoconstriction occurs in OI during orthostasis, which is primarily due to hyperventilation, causing significant hypocapnia. Hypocapnia and symptoms of orthostatic hypertension are reversible by CO2 rebreathing. PMID- 9731614 TI - Early hemicraniectomy in patients with complete middle cerebral artery infarction. AB - BACKGROUND AND PURPOSE: Malignant, space-occupying supratentorial ischemic stroke is characterized by a mortality rate of up to 80%. Several reports indicate a beneficial effect of hemicraniectomy in this situation. However, whether and when decompressive surgery is indicated in these patients is still a matter of debate. METHODS: In an open, prospective trial we performed hemicraniectomy in 63 patients with acute complete middle cerebral artery infarction. Initial clinical presentation was assessed by the Scandinavian Stroke Scale (SSS) and the Glasgow Coma Scale (GCS). All survivors were reexamined 3 months after surgical decompression, with the clinical evaluation graded according to the Rankin Scale (RS) and Barthel Index (BI). We analyzed the influence of early decompressive surgery (<24 hours after symptom onset, based on clinical status at admission and initial CT findings) versus late surgery (>24 hours after first reversible signs of herniation) on mortality, functional outcome, and the length of time of critical care therapy was needed. RESULTS: In total, 46 patients (73%) survived. Despite complete hemispheric infarction, no survivor suffered from complete hemiplegia or was permanently wheelchair bound. In patients with speech-dominant hemispheric infarction (n=11), only mild to moderate aphasia was present. The mean BI score was 65, and RS score revealed severe handicap in 13% of the patients. In 31 patients with early decompressive surgery, mortality was 16% and BI score 68.8. Early hemicraniectomy led to a significant reduction in the length of time critical care therapy was needed (7.4 versus 13.3 days, P<0.05). CONCLUSIONS: In general, the outcome of patients treated with craniectomy in severe ischemic hemispheric infarction was surprisingly good. In addition, early decompressive surgery may further improve outcome in these patients. PMID- 9731613 TI - The apolipoprotein E epsilon4 allele and outcome in cerebrovascular disease. AB - BACKGROUND AND PURPOSE: Polymorphism of the apolipoprotein E gene (APOE) may influence outcome after traumatic brain injury and intracerebral hemorrhage, with the epsilon4 allele being associated with poorer prognosis. We investigated APOE allele distribution in acute stroke and the effect of the epsilon4 allele on outcome. METHODS: APOE genotypes were determined in 714 stroke patients: 640 ischemic stroke and 74 intracerebral hemorrhage patients. The survival effect of the epsilon4 allele was assessed with the use of a stratified log-rank test. A Cox proportional hazards regression model was used to estimate the independent effect of epsilon4 dose (0, 1, or 2) on survival, and logistic regression was used to determine the effect on 3-month outcome (good if alive at home, poor if in care or dead). RESULTS: Allele distribution matched the general population with no difference between the ischemic and hemorrhagic groups. Survival in the entire cohort was unaffected by epsilon4 dose. Improved survival with increasing epsilon4 dose was found in the ischemic group (relative hazard=0.76 per allele; P=0.04). If transient ischemic attacks were excluded, a trend for improved survival persisted (P=0.06). With intracerebral hemorrhage, a trend was seen toward reduced survival with epsilon4 (P=0.07, log-rank test). Three-month outcome in the ischemic group was unaffected by epsilon4 dose, and a trend toward poorer outcome with epsilon4 was seen for intracerebral hemorrhage (P=0.10). CONCLUSIONS: The APOE epsilon4 allele had divergent effects on survival and outcome in ischemic and hemorrhagic strokes in this population. The reported adverse effect on patients with intracerebral hemorrhage was supported. The favorable survival effect on ischemic stroke patients requires further study. PMID- 9731615 TI - Local intra-arterial thrombolysis in acute ischemic stroke. AB - BACKGROUND AND PURPOSE: We performed a retrospective analysis of the prognostic factors in patients treated with local intra-arterial thrombolysis (LIT). The purpose of this study was to evaluate the safety and efficacy of LIT using urokinase in patients with acute ischemic stroke of the anterior or posterior circulation and to determine the influence of clinical and radiological parameters on outcome. METHODS: Forty-three patients were treated with LIT using urokinase (median dose, 0.75x10(6) IU). The median National Institutes of Health Stroke Scale (NIHSS) score at hospital admission was 18 (range, 9 to 36). Nine patients had occlusions of the internal carotid artery (ICA), 23 of the middle cerebral artery (MCA), 1 of the anterior cerebral artery, and 10 of the basilar artery (BA). Outcome was assessed after 3 months and classified as good for Rankin Scale (RS) scores of 0 to 3 and poor for RS scores of 4 or 5 and death. RESULTS: Nine patients (21%) recovered to RS scores 0 or 1, 17 (40%) to scores of 2 or 3, and 7 (16%) to scores of 4 or 5. Ten patients (23%) died. Outcome was good in 17 patients (80%) with MCA occlusions, in 3 patients (33%) with ICA, and in 5 patients (50%) with BA occlusions. Good outcome was associated with an initial NIHSS score of <20 (P<0.001), improvement by 4 or more points on NIHSS score within 24 hours (P=0.001), and vessel recanalization (P=0.02). Recanalization was more likely if LIT was started within 4 hours (P=0.01). Symptomatic cerebral hemorrhage occurred in 2 patients (4.7%). CONCLUSIONS: LIT was most efficacious in patients with MCA and BA occlusions when the initial NIHSS score was less than 20 and when treated within 4 hours. It is of limited value in patients with distal ICA occlusions. PMID- 9731616 TI - Transient response harmonic imaging: an ultrasound technique related to brain perfusion. AB - BACKGROUND AND PURPOSE: Gray-scale harmonic imaging is the first method to visualize blood perfusion and capillary blood flow with ultrasound after intravenous contrast agent application. The purpose of the present study was to evaluate the potential of transient response second harmonic imaging (TRsHI) to assess normal echo contrast characteristics in different brain areas by transcranial ultrasound. METHODS: In 18 patients without cerebrovascular diseases, TRsHI examinations were performed bilaterally with the use of the transtemporal approach after application of 6.5 mL of a galactose-based microbubble suspension (400 mg/mL). The transmission rate was once every 4 cardiac cycles. Regional cerebral contrast was visually assessed and then quantified off-line with the use of time-intensity curves. In 4 different regions of interest (ROI) (posterior part of the thalamus [ROIa], anterior part of the thalamus [ROIb], lentiform nucleus [ROIc], and white matter [ROId]), the following parameters were evaluated: peak intensity, area under the curve (AUC), and time to peak intensity. AUC ratios for ROIc/a, d/a, c/b, and d/b were calculated. RESULTS: In all patients parenchymal contrast enhancement was visually detectable. One hundred thirty-one characteristic time-intensity curves (baseline phase, peak contrast intensity, slow washout phase) were demonstrable in 144 ROIs. In ROIc and ROId, characteristic contrast curves could be observed most frequently (68/72 examinations), whereas time-intensity curves in ROIa and ROIb could not be evaluated because of inadequate contrast enhancement in 9 of 72 examinations. Time to peak intensity varied between 20 and 52 cardiac cycles; in 1 patient it was 88 cardiac cycles. In all individuals AUCs and in 16 of 18 subjects peak intensity in ROIc and ROId showed a 2- to 10-fold increase compared with ROIa and ROIb. In no examination did AUC ratios show a >2-fold side difference irrespective of the ROI. CONCLUSIONS: The present study demonstrates for the first time that TRsHI produces accurate contrast in different brain areas and represents an ultrasonic tool related to brain perfusion. Absolute values of quantitative parameters show high variations caused by different temporal bone thicknesses and a complex relationship between echo contrast concentrations and measurements of optic intensities. Ratios between different ROIs help to compare contrast enhancement in different brain areas. Furthermore, because of the fact that attenuation of contrast enhancement in TRsHI depends strictly on the insonation depth, harmonic imaging studies of brain perfusion cannot be compared directly with other imaging techniques such as positron emission tomography. PMID- 9731617 TI - Endothelial nitric oxide synthase exon 7 polymorphism, ischemic cerebrovascular disease, and carotid atheroma. AB - BACKGROUND AND PURPOSE: The role of endothelial nitric oxide synthase (eNOS) in normal physiology suggests that it could be a potential candidate gene for stroke. Reduced eNOS activity could mediate an increased stroke risk through hypertension or independent of hypertension through abnormal vasomotor responses, promoting atherogenesis, or increased platelet adhesion and aggregation. Recently, a common polymorphism in exon 7 of the eNOS gene (894G-->T) has been reported to be a strong risk factor for coronary artery disease. We determined whether it was also a risk factor for transient ischemic attack (TIA) and ischemic stroke and for carotid atheroma. METHODS: We studied 361 consecutive white patients presenting with ischemic stroke or TIA to a neurological cerebrovascular disease service and 236 normal white controls. In all patients CT and/or MR head imaging and high-resolution carotid duplex ultrasound were performed. The presence of the polymorphism (N/n) was determined by polymerase chain reaction and restriction with the enzyme BanII. RESULTS: There was no difference in the frequency of the NN genotype between patients and controls (13.0% versus 15.3%; P=0.44) or in N allele frequency (39% versus 37%; P=0.57). There was no association with genotype when only patients with stroke (excluding those with TIA) or when only individuals aged < or =65 years were considered. In contrast, there was a highly significant independent association between cerebrovascular disease and hypertension (odds ratio, 2.87; 95% CI, 2.0 to 4.15; P<0.00001), smoking (odds ratio, 2.58; 95% CI, 1.80 to 3.70; P<0.00001), and diabetes (odds ratio, 2.68; 95% CI, 1.38 to 5.24; P=0.004). There was no relationship between the polymorphism and any particular stroke subtype: large vessel disease, for NN, 15 of 105 (14.3%); lacunar disease, 10 of 75 (13.3%); cardioembolic and unknown, 18 of 151 (11.9%); and tandem pathology, 4 of 30 (13.3%) (P=0.68, chi2). There was no difference in the mean degree of carotid stenosis between the 3 genotypes: NN, 31.1% (SD, 27.1); Nn, 30.1% (29.0); and nn, 31.2% (26.3) (P=0.9). There was no association between the NN genotype or the N allele and hypertension. CONCLUSIONS: We failed to find a relationship between this exon 7 polymorphism and ischemic cerebrovascular disease. In particular, it was not associated with stroke and TIA secondary to large-vessel atherosclerosis or with the degree of carotid stenosis in patients with cerebrovascular disease. It is unlikely that this particular polymorphism or any closely linked polymorphism is a major risk factor in the majority of white patients with stroke. PMID- 9731618 TI - Retrograde transvenous neuroperfusion: a back door treatment for stroke. AB - BACKGROUND AND PURPOSE: Stroke is the third leading cause of death and the leading cause of adult disability in the United States. The clot-lysis drug tissue plasminogen activator is the only treatment that has been effective for acute stroke patients, yet there are significant limitations to its use and effectiveness. In this study retrograde transvenous neuroperfusion (RTN) was evaluated for its efficacy in reversing acute ischemia, preventing paralysis, and limiting pathological evidence of infarction in baboons. METHODS: Ten adult male baboons underwent 3.5 hours of reversible middle cerebral artery occlusion (MCAO) under isoflurane (0.25% to 1.5%) anesthesia. Five randomly chosen animals received RTN treatment 1 hour after start of MCAO. Somatosensory evoked potentials were recorded during MCAO. Animals were assigned daily neurological scores. Animals were killed 6 days after MCAO, and brains were quantitatively analyzed for infarct volume. RESULTS: Within 1 hour after RTN was started, treated animals showed significantly improved somatosensory evoked potentials (103.3% versus 75% of baseline; P<0.01). Likewise, the combined neurological score for the RTN-treated group was 99.2, while the combined mean score for the untreated group was 66.4 (P<0.015). The mean infarction volume was 8.8+/-3.1% (of contralateral hemisphere) for the control group and 0.3+/-0.2% for the RTN treated group (P<0.01). No increased mortality was seen in the RTN-treated group. CONCLUSIONS: We conclude that RTN treatment during MCAO effectively reverses the pathophysiological sequelae of ischemia, even when the treatment is initiated 1 hour after the onset of ischemia. Although the infarct volume in the control group was variable when quantitatively assessed 6 days after 3.5 hours of MCAO, virtually no evidence of infarcts was seen in the RTN-treated group. PMID- 9731619 TI - Intracerebral hemorrhage in the rat: effects of hematoma aspiration. AB - BACKGROUND AND PURPOSE: Deep intracerebral hemorrhage is associated with considerable mortality and morbidity, but the value of surgical therapy is debatable. The purpose of this study was to evaluate whether aspiration of the hematoma in a rodent model of intracerebral hemorrhage could improve final neurological outcome. METHODS: Intracerebral hemorrhage was induced in 2 groups of rats by injection of bacterial collagenase into the caudate nucleus. In 1 group of rats, streptokinase was used to lyse the hematoma 4 hours after hemorrhage induction, and the clot was then aspirated. Behavioral function was evaluated repeatedly until the rats were killed 7 weeks after collagenase injection. Histology was used to assess neuronal loss, astroglial proliferation, and overall brain morphology. In a second experiment, brain water was measured at 24 hours. RESULTS: The treated rats performed significantly better than controls on a motor-behavior evaluation on days 1, 2, and 28 after aspiration. Skilled forelimb testing performed for 3 weeks after the global behavior evaluations showed a significant deficit of contralateral forelimb function in both groups, but there was no significant difference between the 2 groups. Neuronal loss in the perihematoma striatum was significantly greater in untreated compared with treated rats. In most rats, structural damage extended into the internal capsule and thalamus. CONCLUSIONS: Aspiration of the hematoma after collagenase-induced hemorrhage slightly improved acute functional outcome and reduced neuronal loss from the striatum. Further studies are required to delineate the mechanism of the effect. PMID- 9731620 TI - Endothelin B receptor antagonists attenuate subarachnoid hemorrhage-induced cerebral vasospasm. AB - BACKGROUND AND PURPOSE: While it has been widely reported that the vasospasm following subarachnoid hemorrhage (SAH) is prevented/reversed by endothelin (ET) receptor antagonists selective for the ET(A) receptor and by nonselective ET receptor antagonists, ie, antagonists of both the ET(A) and ET(B) receptors, there are no reports on the possible attenuation of the spasm by selective ET(B) receptor antagonists. The purpose of this study was to investigate whether (1) ET(B) receptor antagonists prevent and reverse SAH-induced spasm and (2) attenuation of the spasm results from blockade of smooth muscle ET(B) (ET(B2)) receptor-mediated constriction and/or endothelial ET(B) (ET(B1)) receptor mediated ET-1-induced ET-1 release. METHODS: SAH-induced spasm of the rabbit basilar artery was induced with the use of a double hemorrhage model. In vivo effects of agents on the spasm were determined by angiography after their intracisternal infusion (10 microL/h) by mini osmotic pump. In situ effects of agents on the spasm were determined by direct measurement of vessel diameter after their suffusion in a cranial window. RESULTS: SAH constricted the basilar artery by 30%. Intracisternal infusion with 10 micromol/L BQ788, an ET(B1/B2) receptor antagonist, reduced the spasm to 10%. To investigate whether BQ788 prevented the spasm by blockade of ET(B1) receptor-mediated ET-1-induced ET-1 release, as opposed to ET(B2) receptor-mediated constriction, we tested whether ET(B1) receptor blockade also prevented the spasm. Indeed, intracisternal infusion with 10 micromol/L RES-701-1, a selective ET(B1) receptor antagonist, reduced the spasm to 10%. Similarly, in situ superfusion with 1 micromol/L BQ788 reversed the spasm by 40%, and 1 micromol/L RES-701-1 reversed the spasm by 50%. However, both BQ788 and RES-701-1 enhanced by 40% to 50% the 3 nmol/L ET-1 induced constriction elicited in spastic vessels previously relaxed with 0.1 mmol/L phosphoramidon, an ET-converting enzyme inhibitor. CONCLUSIONS: These results demonstrate that ET(B) receptor antagonists prevent and reverse SAH induced cerebral vasospasm in an animal model. The likely mechanism underlying the attenuation of the spasm is blockade of ET(B1) receptor-mediated ET-1-induced ET-1 release of newly synthesized ET-1. These studies provide rationale for the therapeutic use of ET(B1) receptor antagonists to relieve the vasospasm following SAH, as well as other pathophysiological conditions involving possible ET-1 induced ET-1 release. PMID- 9731621 TI - Monoclonal antibodies against ICAM-1 and CD18 attenuate cerebral vasospasm after experimental subarachnoid hemorrhage in rabbits. AB - BACKGROUND AND PURPOSE: Inflammatory responses have been implicated in the elaboration of several forms of central nervous system injury, including cerebral vasospasm after subarachnoid hemorrhage (SAH). A critical event participating in such responses is the recruitment of circulating leukocytes into the inflammatory site. Two of the key adhesion molecules responsible for the attachment of leukocytes to endothelial cells are intercellular adhesion molecule-1 (ICAM-1) and the common beta chain of the integrin superfamily (CD18). This study examined the effects of monoclonal antibodies on ICAM-1 and the effects of CD18 on cerebral vasospasm after SAH. METHODS: A rabbit model of SAH was utilized to test the influence of intracisternally administered antibodies to ICAM-1 and CD18 on cerebral vasospasm. Antibodies were administered alone or in combination, and the cross-sectional area of basilar arteries was assessed histologically on day 2 post-SAH. RESULTS: Treatment with antibodies to ICAM-1 or CD18 inhibited vasospasm by 22% and 27%, respectively. When administered together, the attenuation of vasospasm increased to 56%. All of these effects achieved statistical significance. CONCLUSIONS: These findings provide the first evidence that the severity of cerebral vasospasm can be attenuated using monoclonal antibodies against ICAM-1 and CD18. The results reinforce the concept that cell mediated inflammation plays an important role in cerebral vasospasm after SAH and suggest that therapeutic targeting of cellular adhesion molecules can be of benefit in treating cerebral vasospasm. PMID- 9731622 TI - Ischemic preconditioning and brain tolerance: temporal histological and functional outcomes, protein synthesis requirement, and interleukin-1 receptor antagonist and early gene expression. AB - BACKGROUND AND PURPOSE: A short duration of ischemia (ie, ischemic preconditioning [PC]) can provide significant brain protection to subsequent ischemic events (ie, ischemic tolerance [IT]). The present series of studies was conducted to characterize the temporal pattern of a PC paradigm, to systematically evaluate the importance of protein synthesis in PC-induced IT, and to explore candidate gene expression changes associated with IT. METHODS: Temporary middle cerebral artery occlusion (MCAO) (10 minutes) was used for PC. Various periods of reperfusion (ie, 2, 6, and 12 hours and 1, 2, 7, 14, and 21 days) were allowed after PC and before permanent MCAO (PMCAO) (n=7 to 9 per group) to establish IT compared with non-PC (sham-operated) rats (n=22). Infarct size, forelimb and hindlimb motor function, and cortical perfusion (laser-Doppler flowmetry; n=9 per group) were measured after PMCAO. The effects of the protein synthesis inhibitor cycloheximide administered just before PC (n= 13 to 17) or administered long after PC but just before PMCAO (n=7 to 8) on IT were also determined. Interleukin- receptor antagonist mRNA (reverse transcriptase and polymerase chain reactions [n=20] and Northern analysis [n=50]) and protein expression (immunohistochemistry [n=16]) after PC and early response gene expression (Northern analysis [n=16]) after PMCAO in PC animals were determined. RESULTS: Hemispheric infarct was significantly (P<0.01) reduced only if PC was performed 1 day (decreased 58.4%), 2 days (decreased 58.1%), or 7 days (decreased 59.4%) before PMCAO. PC significantly (P<0.01) reduced neurological deficits (similar to reductions in infarct size). Cycloheximide eliminated ischemic PC induced IT effects on both brain injury and neurological deficits if administered before PC (P<0.05) but not if administered long after PC but before PMCAO. PC did not produce any significant brain injury, alter cortical blood flow after PMCAO, or produce contralateral cortical neuroprotection. Interleukin-1 receptor antagonist mRNA and protein expression were increased significantly (P<0.01) only during the IT period. PC rats also exhibited a significant (P<0.01) reduction in c-fos and zif268 mRNA expression after PMCAO. CONCLUSIONS: PC is a powerful inducer of ischemic brain tolerance as reflected by preservation of brain tissue and motor function. PC induces IT that is dependent on de novo protein synthesis. New protein(s) that occurs at the PC brain site 1 to 7 days after PC contributes to the neuroprotection. Those proteins that are produced after the more severe PMCAO in PC animals apparently do not contribute to IT. The PC-induced IT is also associated with increased expression of the neuroprotective protein interleukin-1 receptor antagonist and a reduced postischemic expression of the early response genes c-fos and zif268. (Stroke. 1998;29:1937-1951.) PMID- 9731623 TI - Delayed treatment with an adenosine kinase inhibitor, GP683, attenuates infarct size in rats with temporary middle cerebral artery occlusion. AB - BACKGROUND AND PURPOSE: Brain ischemia is associated with a marked increase in extracellular adenosine levels. This results in activation of cell surface adenosine receptors and some degree of neuroprotection. Adenosine kinase is a key enzyme controlling adenosine metabolism. Inhibition of this enzyme enhances the levels of endogenous brain adenosine already elevated as a result of the ischemic episode. We studied a novel adenosine kinase inhibitor (AKI), GP683, in a rat focal ischemia model. METHODS: Four groups of 10 adult Sprague-Dawley rats were exposed to 90 minutes of temporary middle cerebral artery (MCA) occlusion. Animals were injected intraperitoneally with vehicle, 0.5 mg/kg, 1.0 mg/kg, or 2.0 mg/kg of GP683 30, 150, and 270 minutes after the induction of ischemia by a researcher blinded to treatment group. The animals were euthanatized 24 hours after MCA occlusion, and brains were stained with 2,3,5-triphenyltetrazolium chloride. We measured brain temperatures in a separate group of 6 rats before and after administration of 1.0 mg/kg GP683. RESULTS: All treated groups showed a reduction in infarct volumes, but a significant effect was observed only in the 1.0 mg/kg-dose group (44% reduction, P=0.0077). Body weight, physiological parameters, neurological scores, and mortality did not differ among the 4 groups. No apparent behavioral side effects were observed. Brain temperatures did not change after drug injection. CONCLUSIONS: Our results indicate that the use of AKIs offers therapeutic potential and may represent a novel approach to the treatment of acute brain ischemia. The therapeutic effect observed was not caused by a decrease in brain temperature. PMID- 9731624 TI - Expression and function of recombinant endothelial nitric oxide synthase gene in canine basilar artery after experimental subarachnoid hemorrhage. AB - BACKGROUND AND PURPOSE: Gene transfer with recombinant viral vectors encoding vasodilator proteins may be useful in therapy of cerebral vasospasm after subarachnoid hemorrhage (SAH). Relaxations mediated by nitric oxide are impaired in cerebral arteries affected by SAH. The present study was designed to determine the effect of SAH on the efficiency of ex vivo adenovirus-mediated gene transfer to canine basilar arteries and to examine whether expression of recombinant endothelial nitric oxide synthase (eNOS) gene may have functional effects on vasomotor reactivity of spastic arteries affected by SAH. METHODS: Replication deficient recombinant adenovirus vectors encoding bovine eNOS (AdCMVeNOS) and Escherichia coli beta-galactosidase (AdCMVbeta-Gal) genes were used for ex vivo gene transfer. Rings of basilar arteries obtained from control dogs and dogs exposed to SAH were incubated with the vectors in minimum essential medium. Twenty-four hours after gene transfer, expression and function of the recombinant genes were evaluated by (1) histochemical or immunohistochemical staining, (2) beta-galactosidase protein measurement, and (3) isometric tension recording. RESULTS: Transduction with AdCMVbeta-Gal and AdCMVeNOS resulted in the expression of recombinant beta-galactosidase and eNOS proteins mostly in the vascular adventitia. The expression of beta-galactosidase protein was approximately 2-fold higher in SAH arteries than in normal arteries. Endothelium-dependent relaxations caused by bradykinin and substance P were suppressed in SAH arteries. The relaxations to bradykinin were significantly augmented in both normal and SAH arteries after AdCMVeNOS transduction but not after AdCMVbeta-Gal transduction. The relaxations to substance P were augmented by AdCMVeNOS transduction only in normal arteries. Bradykinin and substance P caused relaxations even in endothelium-denuded arteries, when the vessels were transduced with AdCMVeNOS. These endothelium-independent (adventitia-dependent) relaxations to bradykinin observed after AdCMVeNOS transduction were similar between normal and SAH arteries, whereas those to substance P were significantly reduced in SAH arteries compared with normal arteries. CONCLUSIONS: These results suggest that expression of recombinant proteins after adenovirus-mediated gene transfer may be enhanced in cerebral arteries affected by SAH and that successful eNOS gene transfer to spastic arteries can at least partly restore the impaired nitric oxide-mediated relaxations through local (adventitial) production of nitric oxide. PMID- 9731625 TI - Behavioral testing does not exacerbate ischemic CA1 damage in gerbils. AB - BACKGROUND AND PURPOSE: Previous research studying ablative lesions has suggested that functional use may exacerbate brain injury. If true, this would have considerable ramifications not only for the mechanistic understanding of neuronal injury but also for the clinical use of physiotherapy. In this report the hypothesis that behavioral use of brain tissue exacerbates ischemic hippocampal injury was tested. METHODS: Gerbils were subjected to sham operation or 5 minutes of normothermic ischemia. To produce borderline hippocampal CA1 injury and enhance susceptibility to exacerbation, 2 of 3 ischemic groups were cooled (>48 hours) beginning at 6 hours after ischemia. Increased use of the hippocampus was produced by a battery of tests involving 3 novel small mazes, a T maze, and an open field. One hypothermic group was not tested and served as a control. RESULTS: Behavioral testing failed to worsen ischemic damage since neuronal loss in the behaviorally tested and untested hypothermic groups was 12% and 8%, respectively, while that in the untreated ischemic group was 81% at a 1-month survival. Accordingly, protected CA1 cells tolerated the neuronal activity associated with behavioral testing. Concomitant with marked CA1 neuroprotection, a significant reduction in behavioral deficits with the hypothermic treatment was observed. Importantly, behavioral testing was found to transiently elevate brain temperature. CONCLUSIONS: CA1 neuronal survival was unaffected by behavioral testing or the associated mild fever. Hypothermia delayed for 6 hours provided sustainable CA1 neuroprotection. PMID- 9731626 TI - Neuronal damage and plasticity identified by microtubule-associated protein 2, growth-associated protein 43, and cyclin D1 immunoreactivity after focal cerebral ischemia in rats. AB - BACKGROUND AND PURPOSE: An objective of therapeutic intervention after cerebral ischemia is to promote improved functional outcome. Improved outcome may be associated with a reduction of the volume of cerebral infarction and the promotion of cerebral plasticity. In the developing brain, neuronal growth is concomitant with expression of particular proteins, including microtubule associated protein 2 (MAP-2), growth-associated protein 43 (GAP-43), and cyclin D1. In the present study we measured the expression of select proteins associated with neurite damage and plasticity (MAP-2 and GAP-43) as well as cell cycle (cyclin D1) after induction of focal cerebral ischemia in the rat. METHODS: Brains from rats (n=28) subjected to 2 hours of middle cerebral artery occlusion and 6 hours, 12 hours, and 2, 7, 14, 21, and 28 days (n=4 per time point) of reperfusion and control sham-operated (n=3) and normal (n=2) rats were processed by immunohistochemistry with antibodies raised against MAP-2, GAP-43, and cyclin D1. Double staining of these proteins for cellular colocalization was also performed. RESULTS: Loss of immunoreactivity of both MAP-2 and GAP-43 was observed in most damaged neurons in the ischemic core. In contrast, MAP-2, GAP 43, and cyclin D1 were selectively increased in morphologically intact or altered neurons localized to the ischemic core at an early stage (eg, 6 hours) of reperfusion and in the boundary zone to the ischemic core (penumbra) during longer reperfusion times. CONCLUSIONS: The selective expressions of the neuronal structural proteins (MAP-2 in dendrites and GAP-43 in axons) and the cyclin D1 cell cycle protein in neurons observed in the boundary zone to the ischemic core are suggestive of compensatory and repair mechanisms in ischemia-damaged neurons after transient focal cerebral ischemia. PMID- 9731627 TI - Photothrombotic middle cerebral artery occlusion in spontaneously hypertensive rats: influence of substrain, gender, and distal middle cerebral artery patterns on infarct size. AB - BACKGROUND AND PURPOSE: To analyze the effects of substrain and gender differences in spontaneously hypertensive rats (SHR) and distal middle cerebral artery (MCA) branching patterns on infarct size, we compared infarct volumes produced by photothrombotic distal MCA occlusion using SHR/Kyushu and SHR/Izumo (Izm). METHODS: Twenty-four male and 8 female SHR/Kyushu, 15 male and 5 female SHR/Izm, and 6 male Wistar-Kyoto rats (WKY)/Izm (5 to 7 months old) were subjected to photothrombotic distal MCA occlusion, and infarct volumes were determined. RESULTS: Although blood pressure levels were essentially the same between the two substrains of hypertensive rats, infarct volumes were significantly larger in the SHR/Kyushu substrain than in SHR/Izm of either sex (P<0.001); infarct volumes in male and female SHR/Kyushu were 83.8+/-11.7 and 58.5+/-9.2 mm3, and those in male and female SHR/Izm were 61.5+/-10.7 and 34.8+/ 7.9 mm3, respectively (values are mean+/-SD). Male SHR/Kyushu that had simple Y shaped MCA showed smaller infarcts (75.8+/-14.6 mm3, n=11) than those with more branching (regular) MCA (93.2+/-19.1, n=13), the difference being significant (P=0.022). Male SHR/Izm with simple distal MCA also produced smaller infarctions than those with regular MCA (51.0+/-3.7 versus 68.9+/-8.7 mm3, P=0.0004). CONCLUSIONS: Photothrombotic occlusion of distal MCA in hypertensive rats provides a simple and reproducible model of focal ischemia. Most importantly, this study emphasizes the substantial variabilities in infarct sizes caused by the differences in substrains of SHR, gender, and distal MCA patterns. PMID- 9731628 TI - Mechanisms of motor dysfunction after transient MCA occlusion: persistent transmission failure in cortical synapses is a major determinant. AB - BACKGROUND AND PURPOSE: Failure of prompt motor recovery after spontaneous recirculation or thrombolytic therapy may be due to an unsatisfactory restoration of synaptic activity within cortex and/or blockade of electrical impulses at the severely ischemic subcortical region. METHODS: Afferent, efferent, and synaptic activities were focally examined within the rat sensorimotor cortex by recording the somatosensory-evoked potential (SEP) and motor area response evoked by stimulation of premotor afferents (PmEP) intracortically and the motor-evoked potential (MEP) generated by stimulation of the forelimb area from the brain stem. The effect of ischemia on electrical activity in the cortex and on axonal conduction in the subcortical region was studied differentially by proximal or distal occlusion of the MCA. RESULTS: MEP consisted of direct and indirect waves generated by direct activation of pyramidal axons and indirect excitation of pyramidal neurons via cortical synapses, respectively. MEP, PmEP, and SEP disappeared on proximal occlusion. Following reperfusion after 1 to 3 hours of ischemia, the direct wave of MEP readily recovered but the indirect wave showed no improvement, suggesting a restored axonal conduction but impaired cortical synaptic transmission. The synaptic defect, which also caused a poor recovery in PmEP and SEP and on electrocorticogram, was persistent and detected 24 hours after 1 hour of proximal occlusion. CONCLUSIONS: Our data suggest that motor dysfunction is caused by loss of cortical excitability and blockade of motor action potentials at the subcortical level during ischemia. After brief transient ischemia, axonal conduction readily recovers; however, a persistent transmission failure at cortical synapses leads to motor dysfunction. PMID- 9731630 TI - Morbidity of intracranial hemorrhage in patients with cerebral arteriovenous malformation. PMID- 9731629 TI - Recurrent intracranial hemorrhage due to postpartum cerebral angiopathy: implications for management. AB - BACKGROUND: Postpartum cerebral angiopathy as a cause of hemorrhagic stroke in young women is not well recognized. It is unknown whether this disorder represents a true inflammatory vasculitis or transient vasoconstriction related to the hormonal events of pregnancy and the postpartum period. CASE DESCRIPTION: A 39-year-old woman presented with postpartum intracranial hemorrhage and, 32 months later, with subarachnoid hemorrhage, following normal pregnancies. Cerebral angiography obtained after each stroke demonstrated diffuse irregularity of branches of the middle cerebral arteries consistent with a diffuse vasospastic process or classic vasculitis. Neurological deficits resolved and results of a transcranial Doppler study normalized after a short course of high-dose corticosteroids following the second stroke. CONCLUSIONS: Postpartum cerebral angiopathy should be considered in the differential diagnosis of recurrent intracranial hemorrhagic stroke in young women. Recognition of this condition may preclude treatment with potentially toxic therapies for vasculitis and will have important implications for counseling women on subsequent pregnancies. PMID- 9731631 TI - Depressed platelet status in an elderly patient with hemorrhagic stroke after thrombolysis for acute myocardial infarction. PMID- 9731632 TI - Comparison between endothelial and neuronal nitric oxide pathways in rat aorta and gastric fundus. AB - This study examines the ability of different nitric oxide synthase (NOS) inhibitors and NO donors to inhibit the endothelium-dependent relaxation of the rat aorta and the NANC relaxation of the rat gastric fundus. NG-Nitro-L-arginine, N-monomethyl-L-arginine, and S-methyl-L-thiocitrulline elicite comparable potency in the aorta and in the fundus. However, 1-(2-trifluoromethyl)imidazole (TRIM), unlike 7-nitroindazole, is more potent on the fundus than on the aorta, showing that TRIM elicits a selective functional inhibition of the neural NOS isoform. (1H)-(1,2,4)Oxadiazole(4,3-a)quinoxalin-1-one, a selective inhibitor of soluble guanylyl cyclase, inhibits the dilator response in both tissues and the cyclic GMP mimetic, 8-Br-cGMP, is 16 times more potent for inducing relaxation in the gastric fundus than in the aorta. However, methylene blue and LY-83583, two other inhibitors of soluble guanylyl cyclase and superoxide anion-generating agents, are at least 100 times less potent on fundus strips than on aortic rings. The data suggest that once released into the extracellular space, NO is more susceptible to inactivation by superoxide anions in the vascular tissue than in the gastric fundus. Thus, the study shows that selective inhibition of NO in a target tissue may be reached not only at the NOS isoform level but also by the manipulation of the NO pathway. PMID- 9731633 TI - Certain S-substituted isothioureas not only inhibit NO synthase catalytic activity but also decrease translation and stability of inducible NO synthase protein. AB - In an attempt to identify potent inhibitors of inducible (type II) NO synthase (iNOS) for use in cell culture systems, we found that two S-substituted isothioureas were very potent in cell culture but one such compound also interfered with the induction of NO synthase. S-Ethylisothiourea (EITU) and S aminoethylisothiourea (AEITU) were found to be much more potent than NG methylarginine, NG-nitroarginine methy lester, or aminoguanidine as inhibitors of NO production by cultured RAW 264.7 cell macrophages activated by lipopolysaccharide (LPS). The approximate EC50 values as inhibitors of NO production, assessed by 24-h accumulation in cell culture media, were 10 microM (EITU), 30 microM (AEITU), 300 microM (NG-methylarginine), and 1000 microM (aminoguanidine). EITU was found to inhibit NO production by activated macrophages without interfering with the induction of iNOS. More specifically, EITU failed to influence transcription of iNOS mRNA (Northern blot analysis), translation of iNOS protein (pulse experiments), or degradation of translated iNOS protein (pulse-chase experiments). In contrast, however, AEITU interfered markedly with the induction of iNOS by mechanisms attributed to inhibition of translation of iNOS mRNA into functional protein as well as acceleration of degradation of already translated iNOS protein. These observations indicate that AEITU should not be used in cell culture experiments where the intent is solely to assess the consequences of inhibition of iNOS catalytic activity. PMID- 9731634 TI - Nitric oxide activates granule-associated DNase in human monocytes. AB - Activated and differentiated human monocytes with a CD14+CD16+ phenotype were found to contain a DNase activity associated with secretion granules. Activated cells were obtained from patients with autoimmune diseases. Activation and differentiation of monocytes were also achieved after incubation of PBMC from healthy subjects with protein A (SpA) or immunopotentiating peptides. DNase activity corresponded to a 66-kDa protein, similar to that described in granules from T lymphocytes, active preferentially on double-strand DNA. DNA fragmentation activity increased when NO donors were present; the activity was higher in the presence of Ca2+, and at low pH values. The Ca2+-dependent activity was inhibited by Zn2+. NO-dependent activity was additive with that of Ca2+-dependent and it was not inhibited by Zn2+. Dithiothreitol did not modify the effect of NO on DNase activity. Incubation of PBMC in the presence of NMLA, an inhibitor of NO synthases, decreased this DNase activity. Data reported clearly suggest a regulatory role of NO in granule-associated DNase activity. PMID- 9731635 TI - Induction of nitric oxide in human monocytes and monocyte cell lines by Mycobacterium tuberculosis. AB - The induction of nitric oxide in human monocytes during mycobacterial infection has been a controversial issue. This study describes a comparative evaluation of the colorimetric and fluorometric methods for the detection of NO in response to Mycobacterium tuberculosis (MTB) infection in human peripheral blood-derived monocytes (PBM) and in U937, a human monocyte-derived cell line. MTB was grown in monocyte cultures in vitro for 7 or 10 days. RPMI 1640 medium, without antibiotics and supplemented with L-arginine, Hepes, 5% human AB serum, and tetrahydrobiopterin was used to support monocyte growth. As early as 72 h after infection, soluble nitrite was detectable in the medium using the fluorometric assay with diaminonaphthalene (DAN). Early induction of NO correlated with an increase in the levels of iNOS mRNA as quantitated by RT-PCR. NO levels increased progressively up to day 10 (PBM) or day 7 (U937), when 150-200 nM/10(6) cells of soluble nitrite accumulated in cultures, as measured by DAN. Furthermore, monocytes stained positively for human iNOS protein and peroxynitrite after infection with MTB. The induction of NO by MTB was inhibited by four different inhibitors of iNOS enzyme including N-monomethylarginine. Inhibition of NO resulted in the enhancement of the intracellular growth of two of five clinical isolates of MTB. NO released from a donor (S-nitroso-N-penicillamine) also had a direct bacteriostatic effect on the same isolates in broth cultures. MTB strains thus showed a differential susceptibility to intracellular and extracellular NO. In most of these assays, the Greiss reagent was limited by its sensitivity and remained negative for soluble nitrite throughout the 7-10 days of incubation. Thus, the colorimetric method, which is widely used, may give false-negative results in NO assays. This report also demonstrates for the first time that MTB induces mRNA for iNOS, iNOS protein, NO, and peroxynitrite in human monocyte/macrophage cultures. PMID- 9731636 TI - Localization of inducible nitric oxide synthase mRNA in inflamed gastrointestinal mucosa by in situ reverse transcriptase-polymerase chain reaction. AB - Immunohistochemistry has been critical in determining the tissue localization of inducible nitric oxide synthase (iNOS). However, this technique suffers from nonspecific staining which may lead to false-positive results and the failure of antisera to recognize iNOS from different species. We developed a technique to determine the localization of iNOS mRNA, as opposed to protein, in tissue sections using an in situ RT-PCR (IS RT-PCR) technique. Sections of inflamed gastrointestinal mucosa were used because they were known to be positive for iNOS. The IS RT-PCR technique localized iNOS mRNA to the same sites as immunoreactive iNOS in human gastritis associated with Helicobacter pylori infection, Crohn's disease, and experimental inflammatory bowel disease induced by the hapten TNBS, in rat colon and in guinea pig ileum. The detection of mRNA had an excellent signal-to-noise ratio. Preservation of tissue morphology was poorer than that with immunohistochemistry due the cycles of heating required in the PCR process. This method could be very useful in detecting iNOS gene expression in situations of excessive nonspecific staining with immunohistochemistry or of failure of antibodies to react because of species differences. This technique is also readily applicable to detect RNA and DNA markers of other disease processes. PMID- 9731637 TI - Synthesis, decomposition, and vasodilator action of some new S-nitrosated dipeptides. AB - A number of amino acid methyl esters have been coupled to N-acetylpenicillamine to give a range of sulfur-containing dipeptides. These have been nitrosated to give a family of structurally related NO-donor drugs. The catalytic effect of copper ions upon the release of NO from these compounds is much less than that upon S-nitroso-N-acetylpenicillamine. However, all the nitrosated dipeptides respond in a similar way with little variation in the value of kCu. On the other hand, the vasodilator action of these compounds and the inhibiting effect of hemoglobin do vary quite considerably within the family. It is suggested that this indicates some tissue penetration by these drugs. PMID- 9731639 TI - Pulmonary arteriovenous fistulas in patients with left isomerism and cardiac malformations. PMID- 9731640 TI - The role of devices in the closure of atrial septal defects in the oval fossa. PMID- 9731638 TI - Human and rat neutrophils constitutively express neural nitric oxide synthase mRNA. AB - Freshly isolated rat circulating neutrophils (PMN) constitutively expressed neural nitric oxide synthase (nNOS) mRNA and nNOS protein and exhibited spontaneous basal release of low concentrations of nitrate and nitrite anion (RNI). In contrast, rat peripheral monocytes and macrophages were devoid of nNOS mRNA and protein and did not exhibit basal or spontaneous release of RNI. Constitutive neural NOS mRNA was also found in human PMN. However, nNOS protein was not expressed and spontaneous generation of RNI was absent in the human PMN. Spontaneous release of RNI from rat PMN was inhibited by 7-nitroindazole but not by L-N-iminoethyllysine, which further supported the nNOS origin of the spontaneously produced RNI. Intravenous administration of Escherichia coli endotoxin (0.6 mg/kg) did not acutely affect the content of nNOS mRNA or protein but inhibited nNOS-derived production of RNI in PMN and up-regulated iNOS mRNA and iNOS protein in PMN, macrophages, and monocytes. This communication demonstrates the existence of nNOS mRNA in rat and human PMN and nNOS protein in rat PMN. Moreover, the data also show that the nNOS system in rat PMN is functional and is inhibitable by the nNOS inhibitor 7-nitroindazole. These findings offer an explanation for the spontaneous formation of the PMN-derived relaxing factor resembling nitric oxide (NO). Moreover, since basal production of NO can affect expression of adhesion molecules and cell-cell binding, the nNOS system within the rat may play an important role in PMN function in normal and disease states. Finally and speculatively, if constitutively expressed nNOS mRNA is subject to activation and translation into nNOS protein, nNOS may also play a role in the function of human PMN. PMID- 9731641 TI - Commentary on social research about young people with congenital conditions. PMID- 9731642 TI - Pulmonary and systemic arteriovenous fistulas in patients with left isomerism. AB - Hepatic venous blood has been thought to play some role as a vasoactive agent in the development of pulmonary arteriovenous fistulas in patients with congenital heart disease. During the last 15 years, we have observed pulmonary arteriovenous fistulas in 3, and systemic arteriovenous fistulas in 2, patients from our 16 cases of left isomerism. During the same period, neither pulmonary nor systemic arteriovenous fistulas were detected among 50 patients with right isomerism. Pulmonary arteriovenous fistulas had developed in the absence of surgery in 1 of the patients. Both pulmonary and systemic fistulas were detected in an another patient, in whom the hepatic venous blood bypassed the pulmonary circulation. The level of somatostatin, which is known to reduce splanchnic blood flow, was high in the systemic venous blood of this patient. Although the mechanism of development of the fistulas has yet to be clarified, we should be aware that not only pulmonary, but also systemic arteriovenous fistulas can be found in patients with left isomerism, even prior to any surgical intervention. PMID- 9731643 TI - Early clinical experience with use of the 'Amplatzer Septal Occluder' device for atrial septal defect. AB - Device closure of oval fossa atrial septal defects with the Amplatzer Septal Occluder was performed in 26 patients ranging in age from 0.89 to 60.44 years. In eight additional patients no device implant was performed because of the presence of multiple defects or because the defect was of a size unsuitable for closure with the devices currently available. The stretched diameter of the defects that were closed ranged from 4 to 23 mm (mean 14+/-5.4 mm) and device sizes ranged from 4 to 24 mm. Two devices were unstable, of which one embolized to the right atrium after release. Both devices were retrieved at the same procedure. One of these patients subsequently underwent a successful device closure of his defect using a larger (24-mm) device. Three patients had multiple defects, which were successfully closed with a single device. At 1-month follow-up 23/26 (88%) and at 3-month follow-up 22/24 (92%) patients had complete closure of their defects, while two had residual shunts. One further patient who had complete closure of his defect at 1-month post-implant had his device removed and his atrial septal defect patched surgically 8 weeks after device closure. This was done as a result of the development of a vegetation affecting the device after an episode of septicaemia, which was not related to the cardiac problems. There was no procedure-related morbidity or mortality and all patients remain well at the present time. PMID- 9731644 TI - Growing up with congenital heart disease: the dilemmas of adolescents and young adults [see comment]. AB - Advances in diagnosis, medical management and surgical intervention have improved the longevity and quality of life for children with congenital heart disease. Despite this, research studies specifically examining the psychosocial concerns of adolescents and young adults with congenital heart disease are few. To explore the subjective experiences and dilemmas of this population during the transition from adolescence to young adulthood, we interviewed, using a semi-structured protocol, a convenience sample of nine adolescents and young adults. Using analytic procedures inherent in Grounded Theory methodology, seven themes were identified: the dilemma of normality; dilemmas in disclosure; dilemmas in strategies for management of illness; the challenge of social integration versus social isolation; the challenge of dependence versus independence; the challenge of uncertainty; and strategies for coping. An understanding of these experiences by health professionals can be beneficial in helping this clinical population as they grow up and face the challenges of an uncertain, yet promising, future. PMID- 9731645 TI - Verapamil therapy in infants with hypertrophic cardiomyopathy. AB - We sought to evaluate the safety and efficacy of acute and chronic treatment with verapamil in infants with hypertrophic cardiomyopathy. Prior studies have shown an improvement in adults with hypertrophic cardiomyopathy who were treated with verapamil. Acutely, it reduced the degree of left ventricular outflow tract obstruction. Chronic therapy was associated with an improvement in symptoms and increased long-term survival. To date, no data are available on the efficacy of this drug in infants with hypertrophic cardiomyopathy. We evaluated prospectively the safety and efficacy of verapamil in infants. The acute and chronic effects of verapamil on infants with an echocardiographic diagnosis of hypertrophic cardiomyopathy were evaluated at a single institution between 1980 and 1994, with long-term follow-up available until 1996. Acute effects of an intravenous bolus of 0.1 mg/kg and infusion at 0.007 mg/kg/min were evaluated, where possible, in the cardiac catheterization laboratory. Oral verapamil at 3-5 mg/kg/day was started after catheterization. Follow-up included serial clinical, echocardiographic and Holter monitoring. A total of 22 patients were studied, 17 having a presumed diagnosis of primary hypertrophic cardiomyopathy including three with Noonan's syndrome. Acute infusion of the drug was well tolerated by all, without adverse electrophysiological effects. Haemodynamic effects were consistent with a negative inotropic action with significant falls in cardiac index (4.6+/-1.2 to 4.1+/-0.9 l/min/m2), systolic blood pressure (88+/-16 to 82+/ 14 mmHg) and gradient across the left ventricular outflow tract (nine patients 48.2+/-30.4 to 28.4+/-24.1 mmHg). End-diastolic pressure was unchanged (14.0+/ 6.8 to 13.9+/-4.7 mmHg). Three patients with primary hypertrophic cardiomyopathy died (two while being treated). In the group with primary hypertrophic cardiomyopathy continuing with long-term treatment, follow-up revealed regression in two, progression in three (two died) and stability in 10. For those treated, there was a trend towards improvement in clinical status. Verapamil is well tolerated acutely in infants with hypertrophic cardiomyopathy. The outcome was considerably better in these patients compared with prior reports, though careful long-term assessment is needed. PMID- 9731646 TI - Left ventricular mechanics after closure of ventricular septal defect: influence of size of the defect and age at surgical repair. AB - To evaluate the influence of the size of the defect and the age of surgical repair on left ventricular mechanics, including geometry, shape, diastolic and systolic function as well as myocardial contractility, we used cross-sectional echo-Doppler to study 20 patients (12 males, 8 females) who had undergone successful surgical closure of a ventricular septal defect. The patients were divided in two groups, corrected early and late, on the basis of the degree of left-to-right shunting (ratio of pulmonary to systemic output of greater or less than 2.5/1) and the age at the surgical repair (older or younger than 2 years of age). The group undergoing early correction included 11 patients, mean age 7.1+/ 1.8 years (range 4.2-11.8 years), having surgery at mean age of 1.3+/-0.6 years for a large ventricular septal defect (mean ratio of pulmonary to systemic output of 3.1/1; range 3.4-2.7/1) with a mean postoperative follow-up 4.6+/-1.9 years. The group of nine patients undergoing late correction had a mean age of 11.3+/ 4.9 years (range 6.7-17.2 years), with a later surgical repair (mean age 4.7+/ 2.7 years) for a moderate-sized ventricular septal defect (mean pulmonary/systemic output ratio 2.1/1; range 2.3-1.7) and a mean postoperative follow-up of 7+/-4.2 years. Each group of surgically repaired patients was compared with a control group matched for age, body surface area and gender. No significant differences were found between the normal controls and those undergoing early correction for any assessed functional index regarding left ventricular geometry (normalized volumes and mass for body surface area, mass/volume and thickness/radius ratios), shape (long axis-short axis ratio), diastolic (mitral and pulmonary venous flow patterns) and systolic (fractional shortening and rate-corrected mean velocity of circumferential fibre shortening) function. In addition, the data points for each patient for the rate-corrected mean velocity of circumferential fibre shortening to end-systolic stress relationship were within the 95% confidence limits of normal, suggesting normal left ventricular contractility. On the other hand, the patients undergoing surgery at a later age showed a persistent increase of the normalized left ventricular end-diastolic volume and mass, with an higher mass/volume ratio and reduced end-systolic stress compared with normal controls. Furthermore, left ventricular shape (long axis-short axis ratio) was abnormal at end-diastole but with its normal values at end-systole. Our data suggest that, in the presence of a large ventricular septal defect, early successful surgical repair <2 years of age results in complete recovery of left ventricular mechanics in the postoperative follow-up. In contrast, surgical closure at > 2 years of age, even for a moderately sized ventricular septal defect, deleteriously affects postoperative left ventricular geometry and shape. Since prolonged volume overload may be detrimental to myocardial function, earlier surgical repair should be recommended. PMID- 9731647 TI - Epidemiology and diagnosis of ventricular septal defect in Malta. AB - Malta is a small island with minimal changes in its population, making it an ideal location for epidemiological and historical studies dealing with congenital heart malformations. Ventricular septal defect was studied retrospectively from 1930 to 1994. A sharp and significant decline in age at diagnosis was found, predating echocardiography. All defects are now diagnosed by echocardiography, which has resulted in an increased prevalence of this lesion as seen at birth, particularly of minor defects. The prevalence of ventricular septal defect from 1990 to 1994 was 3.85/1000 live births, with 3.03/1000 not needing surgery and 0.83/1000 requiring operative intervention. Half the defects closed spontaneously. The majority of ventricular septal defects overall, and those spontaneously closing, were muscular defects. The overall prevalence at birth was significantly higher than that reported in recent studies using similar methodologies, implying that the reported rate is more likely to be the true prevalence at birth of clinically detectable defects. PMID- 9731648 TI - History, diagnosis, surgery and epidemiology of pulmonary stenosis in Malta. AB - The Maltese population constitutes an ideal location for epidemiological and historical studies dealing with congenital heart malformations. Pulmonary stenosis was studied retrospectively from 1943 to 1994. A sharp, significant decline in age at diagnosis was found, which predates the introduction of echocardiography. All defects are now diagnosed by echocardiography, which has resulted in an increased prevalence at birth of this lesion, particularly of pulmonary stenosis not requiring intervention. The prevalence at birth of pulmonary stenosis from 1990 to 1994 was 1.65/1000 live births (95% CI: 1.21 2.24), with 1.11/1000 mild lesions (95% CI: 0.76-1.62) and 0.54/1000 lesions requiring intervention (95% CI: 0.31-0.92). The prevalence at birth overall was significantly higher than that reported in recent studies with similar methodologies due to the higher pickup of milder variants of pulmonary stenosis by echocardiography. Significantly more pulmonary stenosis was found in females than in males. PMID- 9731649 TI - Rare forms of isolation of the subclavian artery: echocardiographic diagnosis and surgical considerations. AB - Isolation of the subclavian artery is an unusual anomaly in which the subclavian artery arises not from the aortic arch but from a pulmonary artery via an arterial duct. Such isolation most often occurs with a right aortic arch, and in lesions frequently associated with a right arch, such as tetralogy of Fallot. Since 1994, we have undertaken surgery in four young infants with isolated subclavian arteries and unusual associated anomalies, including one with atrioventricular septal defect and common valvar orifice, two with interruption of a left aortic arch and one with interruption of a right aortic arch. In both patients with interrupted left arch, the isolated subclavian artery was diagnosed preoperatively by echocardiography. We emphasize the significant surgical issues. PMID- 9731650 TI - Predictors of developmental outcomes in children with complete transposition. AB - Cognitive, functional, educational achievement and behavioural measures were employed to assess neurobehavioral status in 57 of 60 participants who were initially enrolled in the Baltimore-Washington Infant Study, and who survived surgical correction of complete transposition (concordant atrioventricular and discordant ventriculo-arterial connections). Charts were reviewed to investigate the relationship between birth variables, surgical strategy and developmental outcomes. Higher preoperative weight was associated with better outcomes on the Stanford-Binet Short-term Memory subtest, while lower preoperative oxygen tension was associated with better outcomes on the Abstract/Visual Reasoning subtest and a test of Visual-Motor Integration. Longer total bypass time was associated with poor outcomes on the Short-term Memory subtests. Higher average flow rates during cooling and rewarming were associated with higher scores in the test of short term memory but poorer outcomes on a test for visual motor integration. Longer cooling times were associated with higher scores on the test for Visual-Motor Integration. Patients suffering seizures scored lower on the Stanford-Binet Composite, as well as in their tests of achievement. The data indicate that non verbal skills may be particularly sensitive to variations in surgical strategies employed to correct complete transposition. Overt neurological events, such as seizures, were related to global deficits in intellectual functioning. Prospective studies evaluating systemic variations in surgical procedures and attempts to prevent and manage perioperative neurological events are important for further investigation of neurodevelopmental outcomes in children surviving surgical correction. PMID- 9731651 TI - Possible sources of right-to-left shunting in patients following a total cavopulmonary connection. AB - Despite a good haemodynamic result, many children have a mildly decreased arterial-oxygen saturation following a total cavopulmonary connection. Our study was performed to determine possible mechanisms of right-to-left shunting in these patients. We performed elective cardiac catheterization in 19 children at a mean interval of 3.6 years following a total cavopulmonary connection. The intrapulmonary right-to-left shunt, the intracardiac right-to-left shunt and the total right-to-left shunt were calculated under mechanical ventilation with 100% oxygen. The intrapulmonary right-to-left shunt was 10.8+/-3.5% of the pulmonary blood flow, and the total right-to-left shunt accounted for 18.9+/-5.2% of the systemic blood flow. The intracardiac right-to-left shunt in patients with no relevant venovenous collaterals or leaks in the atrial tunnel was calculated at 6.4+/-3.0% of the systemic blood flow, while the intracardiac right-to-left shunt in patients with relevant collaterals or leaks accounted for 13.0+/-5.9% of the systemic blood flow. Since intrapulmonary arteriovenous fistulas were not demonstrated angiographically in any of our patients, the intrapulmonary right-to left shunt is probably due to low ratios of perfusion to ventilation in some pulmonary segments. The intracardiac right-to-left shunt was due to leaks across the interatrial baffle, collaterals between systemic and pulmonary veins, and to the coronary sinus draining to the pulmonary venous atrium. PMID- 9731652 TI - New approach in partial cavopulmonary connection. AB - The development of pulmonary arteriovenous fistulas after bidirectional cavopulmonary operations, such as the bidirectional Glenn shunt and Kawashima's procedure, has raised concern. Development of these fistulas, which are more frequent than initially thought, can represent a limiting factor in the late outcome of these patients and may even limit the indication for these types of surgery. Whether the fistulas can be reversed by transforming the surgical procedures has yet to be established. In the hope of avoiding this kind of complication, thought to be caused by the lack of passage of a hypothetical hepatic factor through the pulmonary circulation, we have developed an inverted type of bidirectional cavopulmonary connection in which the blood coming from the liver perfuses immediately both lungs. This is made possible by shunting via an intra-atrial tunnel the blood from the superior caval vein directly to the left atrium, and the blood from the inferior caval vein to the right branch of the pulmonary trunk (keeping its bifurcation intact). We describe findings in two patients undergoing successful surgery with this technique. Serial follow-up with contrast echocardiography did not show evidence of arteriovenous pulmonary fistulas. Despite our numbers being small, and the time of follow-up being limited, we believe that it is important to document these and similar cases. PMID- 9731653 TI - Impact of dynamic 3D transoesophageal echocardiography in the assessment of atrial septal defects and occlusion by the double-umbrella device (CardioSEAL) AB - Occlusion of the atrial septal defects in the oval fossa by interventional catheterization has progressed, but still has limitations. Three-dimensional (3D) echocardiography can provide unique views unavailable by cross-sectional imaging. The objective of this study was to define the clinical application of 3D echocardiography in the assessment and monitoring of transcatheter occlusion of atrial septal defects. Three-dimensional echocardiography was attempted prior to occlusion of atrial septal defects in 41 patients (median age 8.6 years). Serial cross-sectional images were acquired by multiplane transoesophageal echocardiography and displayed by means of computer reconstruction. Dynamic 3D echocardiographic images of defects in the oval fossa were obtained in 40 of 41 patients (98%). Volume-rendering demonstrated the anterosuperior rim in 36 (90%) and the inferoposterior rim in 24 (60%), but failed to reveal small additional fenestrations in six. Sizes measured by 3D echocardiography were significantly larger than those provided by cross-sectional transoesophageal echocardiography (p=0.007), but differed little from those obtained with balloon sizing (p=0.6). After occlusion, 3D echocardiography showed positions of all arms of the device in 20 of 24 cases. Location of any protruding arms, or residual defects, were also clearly revealed. Three-dimensional images obtained in 12 patients during deployment of the double-umbrella device were useful in monitoring its position (single-frame) and for explaining the mechanism of protrusion. Current 3D echocardiography provides clinically relevant information for selection of patients for closure of atrial septal defects by interventional catheterization and when monitoring during implantation. Information obtained by this technique can clarify the mechanism of deployment of the device and closure of the defect, therefore influencing outcomes. PMID- 9731654 TI - Radiofrequency catheter ablation of multiple haemodynamically unstable ventricular tachycardias in a patient with surgically repaired tetralogy of Fallot. AB - A patient with repaired tetralogy of Fallot presented with recurrent syncope and had multiple haemodynamically unstable ventricular tachycardias unresponsive to antiarrhythmic medications. Ventricular tachycardias became haemodynamically tolerated with amiodarone, procainamide and dopamine, permitting activation and entrainment mapping. Radiofrequency ablation of three tachycardia circuits was performed. Ventricular tachycardia could not be induced 1 week, and 3 and 9 months later. Radiofrequency ablation is feasible for multiple, haemodynamically unstable ventricular tachycardias in repaired tetralogy of Fallot. PMID- 9731655 TI - Sudden death 10 years after patch aortoplasty for coarctation. AB - A 25-year-old man died 10 years after a Dacron patch was used to repair a coarctation of the aorta. Death was due to rupture of an unrecognized aneurysm at the site of the patch. After the initial operation at the age of 15 years, there had been no signs of residual or recurrent obstruction. He had no evidence of hypertension and was discharged some years later from regular hospital follow-up to the care of his general practitioner. We strongly recommend that patients who have undergone repair of aortic coarctation by patch aortoplasty should have lifelong follow-up in cardiac units with imaging facilities for monitoring aortic dilation. We would now recommend surgical intervention in the presence of progressive aortic dilation. PMID- 9731656 TI - Serious sequels of Kawasaki disease. AB - A male infant, aged 2 month, with Kawasaki disease had a myocardial infarction despite intravenous infusions of gamma globulin and aspirin at high dosage. He developed progressively a thin walled, dilated aneurysm of the apex of the left ventricle which became lined with thrombus despite treatment with warfarin. Another boy, aged 6 years, was noted on the 10th day of the evolution of Kawasaki disease to have developed a giant aneurysm of the main stem of the left coronary artery. Despite infusion of gamma globulin, the aneurysm remained unaltered and developed a thrombus. The thrombus resolved following treatment with warfarin, though the giant aneurysm has persisted. These two cases illustrate the serious consequences that can follow Kawasaki disease despite management optimal by current standards. PMID- 9731657 TI - Delayed hemopericardium due to trivial chest trauma. AB - We report two cases of hemopericardium occurring in seven-month-old and 12-year old boys, who had no history of major trauma. The possible cause of the hemopericardium for the infant was falling from a bed which was 75 cm high two weeks prior to the admission. The 12-year-old boy had fallen from a chair and damaged his chest 4 weeks previously. Their coagulation tests were all normal. By means of pericardiotomy, we drained 120 ml and 1200 ml of blood, respectively. The boys have now been well over follow-up periods of 24 and 18 months, respectively. PMID- 9731658 TI - Resonance imaging of a ruptured aneurysm of the sinus of Valsalva. AB - Ruptured aneurysm of an aortic sinus of Valsalva is a rare cause of left-to-right shunting. We show how resonance imaging can be used to make the diagnosis. This technique can successfully characterize the shunt as well as determine the presence of associated anomalies, such as ventricular septation and aortic regurgitation. It may be the only study required prior to therapeutic intervention. PMID- 9731659 TI - Complete transposition in a chick embryo demonstrated by scanning electron microscopy. AB - Chick embryos are frequently used as animal models when researching the developing heart. In the past, every attempt to induce complete transposition (the combination of concordant atrioventricular and discordant ventriculo arterial connections) failed in chicks, suggesting that it might be impossible to develop a chicken model for this malformation. We demonstrate, to the best of our knowledge, the first well-documented case of complete transposition occurring in the chick. PMID- 9731660 TI - Counselling strategies for parents of infants with congenital heart disease. AB - Congenital heart disease is a significant cause of morbidity and mortality in the newborn. Its diagnosis may lead to a crisis in the affected families; there are the perceived implications of having an abnormality of so vital an organ. To that may be added the assumed guilt or blame, grief and at times anger, frequently experienced by parents of abnormal infants. It often befalls the paediatric cardiologist to initiate counselling while providing the expert information concerning the abnormality and its optimum management. Such counselling differs from that needed for minor lesions as compared for more complex abnormalities where a fatal outcome may ensure. While it is important to provide an accurate diagnosis and management plan to the parents, early detailed information is often confusing and may not be assimilated at a time of great stress. The parents seem more concerned as to whether the infant will survive, what the long term outlook will be, whether he or she will attend school, play, work and so on. With the more severe cardiac abnormalities, especially where there is a family history, one need be aware of the often perceived guilt of the parents. At times, it may be necessary to help the parents retain sufficient 'self-control', delaying the grieving process to enable them to contribute to the decision making. Where the infant has died, a follow-up appointment can facilitate grieving and help deal with unresolved issues. Through skilled counselling, the cardiologist in addition to his/her diagnostic and management skills, may meaningfully influence the ongoing care of the infant. They may help avoid the development of unrealistic fears or an over-optimistic outlook, thereby fostering the normal development of the child. PMID- 9731661 TI - The Brock procedure (closed infundibular resection) for Fallot's tetralogy: 43 years later. AB - Innovative cardiac surgery ('blind' or indirect infundibular resection) for tetralogy of Fallot on a child of 4 years was followed by survival for 43 years without further surgery. The patient remained well until about one year before death, when he developed clinical features of progressive biventricular failure associated with pulmonary hypertension and incompetence of the pulmonary and tricuspid valves. Postmortem examination showed severe damage to one of the leaflets of the pulmonary valve, interpreted as due to inadvertent avulsion during the original surgical procedure. A large ventricular septal defect was present, but there was no residual subpulmonary infundibular obstruction. PMID- 9731662 TI - Right aortic arch with coarctation proximal to the right subclavian artery and Kommerell's diverticulum. PMID- 9731663 TI - The Association for European Paediatric Cardiology: paediatric cardiology and the European Union. PMID- 9731664 TI - Powerlifting versus weightlifting. PMID- 9731666 TI - Controversial E & M coding guidelines deferred. PMID- 9731665 TI - Laxity versus instability. PMID- 9731667 TI - Red blood cell transfusion practices in patients undergoing orthopedic surgery: a multi-institutional analysis. AB - This retrospective review analyzed and compared transfusion practices in patients undergoing orthopedic surgery in five Massachusetts hospitals with current practice guidelines; opportunities for improvement were identified. Patient specific clinical information and data about transfusion practices were obtained from the medical records of 384 Medicare patients undergoing orthopedic surgery between January 1992 and December 1993. The number of patients who donated autologous blood preoperatively differed significantly among hospitals as did the number of autologous units that were unused. The number of blood units transfused at each transfusion event also differed significantly; some surgeons transfused > or =2 units in the majority of their patients, while others transfused 1 unit at a time. This variation in practice was not explained by differences in patients' clinical status. The mean pretransfusion hematocrit was higher for autologous versus allogeneic blood, suggesting more liberal criteria to transfuse autologous blood. Nearly half of all transfusion events were determined to have been potentially avoidable. Avoidable transfusions were also three to seven times more likely with autologous than with allogeneic blood. Significant inter-hospital differences existed in the number of elective surgery patients exposed to allogeneic blood. The major determinant of allogeneic blood use in these patients was the availability of autologous blood. Each additional autologous blood unit available decreased the odds of allogeneic blood exposure twofold. Differences in intraoperative and postoperative blood salvage use also were noted. These findings indicate that significant variations in practice exist. Comparative data enabled hospitals to identify and target specific areas for improvement. PMID- 9731668 TI - An analysis of hylamer and polyethylene bearings from retrieved acetabular components. AB - Hylamer and conventional polyethylene acetabular liners of the same design, revised for a variety of reasons, were examined and compared to assess the performance of Hylamer as a bearing material. Clinical damage modes, linear wear rates, oxidation levels, and mechanical properties were measured. In both series, many liners were retrieved for dislocation. Wear/osteolysis was the most common reason for retrieval in the Hylamer series, while none of the conventional polyethylene liners were retrieved for this reason. Nearly all liners exhibited abrasion, burnishing, scratching, and creep. The Hylamer liners had more cracking, delamination, and pitting. The Hylamer liners had an average linear wear rate of 0.32 mm/year, while the conventional polyethylene liners had an average wear rate of 0.20 mm/year. Due to sample size, no statistical difference in wear rate was noted between the two groups. In general, both the Hylamer and conventional polyethylene showed oxidation peaks subsurface, resulting from their exposure to gamma radiation in air. Liners with elevated oxidation had decreased ultimate tensile strength, elongation, and toughness. For given oxidation levels, the corresponding mechanical properties of Hylamer appeared lower than those of conventional polyethylene. The ultimate tensile strength values ranged from 14 to 33 MPa for Hylamer and 19 to 32 MPa for conventional polyethylene. Elongation ranges were 19% to 350% (Hylamer) and 80% to 375% (conventional). The Hylamer retrievals in this study gave initial indications of performance; Hylamer appeared to behave similarly, but not superiorly, to conventional polyethylene, in the early functional period with respect to clinical wear and clinical performance. Both Hylamer and conventional polyethylene liners were degraded by gamma sterilization in air, with Hylamer liners demonstrating greater property changes. PMID- 9731669 TI - Functional outcome of arthrodesis for failed total knee arthroplasty. AB - To establish the ability of a salvage procedure to restore an independent lifestyle, the SF-36 functional outcome instrument and the pain, mobility, and physical activity subscales of the Arthritis Impact Measurement Scale (AIMS) was used to assess patient function. Nine patients (10 knees) who had undergone arthrodesis for failed total knee arthroplasty were compared with a control group of successful primary total knee arthroplasty patients. Average clinical follow up was 42 months (minimum: 24 months). For six of the eight SF-36 categories, the average scores for the arthrodesis and arthroplasty groups were similar. The average global scores for the two groups were nearly identical. The arthroplasty patients scored better on the AIMS physical activity and mobility subscales than the arthrodesis group, although the latter group fared better on the pain subscale. Overall, global scores favored the arthroplasty patients. The only subscales to show a statistically significant difference between the arthrodesis and arthroplasty groups were the SF-36 physical functioning and the AIMS physical activity subscales. This pilot study demonstrated the ability of a salvage procedure to allow for an independent lifestyle with minimal complications. Furthermore, despite its popularity, the SF-36 does not appear as sensitive as the AIMS to differences in functional status or health outcomes between total knee arthroplasty and arthrodesis patients. PMID- 9731670 TI - Initial stability comparison of modular hip implants in synthetic femurs. AB - Synthetic femurs were used to assess the initial bone-implant interface stability of three total hip systems: Wright Medical Technology's Infinity smooth trochanteric module (S-TM), Infinity porous-coated trochanteric module (PC-TM), and Johnson and Johnson S-ROM with a porous surface. The hips were implanted into synthetic femurs, rigidly fixed, and subjected to internal rotation and cyclic, axial compressive loads. The results showed that all three implants achieved good initial implant stability and would be expected to permit bone ingrowth. The porous-coated implants showed greater initial implant stability with less axial micromotion compared with the smooth implants. This finding suggests that surface texture plays a role in initial stability of uncemented prostheses if the bone behaves similar to the material used in this study. PMID- 9731671 TI - Overuse tendinitis of the intrinsic muscles. AB - Repetitive strain injuries are currently the leading cause of occupational illnesses. This report describes seven patients who presented with the sole symptom of hand pain and subsequently were diagnosed with intrinsic tendinitis. Six of the 7 patients were given injections of a local anesthetic and steroid solution into the region of the lumbrical tunnels for both diagnostic and therapeutic purposes. All 6 patients had immediate short-term resolution of their symptoms in the office, thereby confirming the diagnosis. Complete resolution of symptoms after both the injection and other treatment interventions occurred in 4 patients, and partial resolution of symptoms occurred in the remaining two patients. Follow-up ranged from 3 to 20 months for 6 patients, and the seventh patient was lost to follow-up. The clinical presentation, diagnostic work-up, and treatment of intrinsic tendinitis are described. PMID- 9731672 TI - Migration measurement of cementless acetabular components: value of clinical and radiographic data. PMID- 9731673 TI - Elbow arthrosis as an initial presentation of acromegaly. PMID- 9731674 TI - Legg-Calve-Perthes disease in a 17-month-old child: a 20-year follow-up. PMID- 9731675 TI - Chronic displaced medial epicondyle fracture. PMID- 9731676 TI - Answer please. Hydatid disease of the spine. PMID- 9731677 TI - Informed consent for obesity surgery. AB - A patient cannot consent to an operation without being adequately informed. There are a number of different operations in use today for treatment of severe obesity. The variations are designed to (1) limit food intake and/or (2) create malabsorption. The surgeon has a duty, according to the law of informed consent, to provide all of the information necessary for a reasonable person to decide whether to consent to the operation recommended. Changes in anatomy, function and risk therefore need to be explained. When only limitation of intake is planned, as in vertical banded gastroplasty (VBG), the patient should know how large the pouch will be and how the outlet will be stabilized. When both intake restriction and malabsorption are planned, as in Roux-en-Y gastric bypass (RGB), or biliopancreatic diversion (BPD), the patient should know whether there will be a larger pouch (less restriction) and a short common channel (more malabsorption) as in BPD or a smaller pouch and less malabsorption. Patients should know that if they have an operation that uses maximum malabsorption to bring weight to a nearly normal level, the risk of malnutrition will be increased, which may require further hospitalization and possible operative treatment. When the duodenum is to be bypassed, the patient should know that this will impair iron and calcium absorption, and that access to this area for radiologic and endoscopic procedures may not be possible. Simple drawings can be used to explain what is planned and how the operation will determine body weight, side-effects, and risk. PMID- 9731679 TI - An adjustable vertical banded gastroplasty does not eliminate the risk of staple line disruption. AB - BACKGROUND: The two main reasons for reoperation after vertical banded gastroplasty (VBG) in the treatment of obesity are staple-line disruption and stomal stenosis. PATIENTS: Seven morbidly obese patients of mean (+/-SEM) body mass index (BMI) 43.7 +/- 1.9 kg/m2 treated with an adjustable vertical banded gastroplasty (AVBG). RESULTS: No complications of the band system were reported. Weight-loss [BMI at 2 years follow-up 33.9 +/- 6.9 kg/m2 (n = 5)] was equivalent to that seen after VBG with a fixed band. Two of the patients developed staple line disruption at 18 and 24 months after surgery. CONCLUSION: AVBG allows adjustment of the stoma, but staple-line disruption was common in this small series. It is possible that an excessive filling of the band in order to achieve excess weight loss results in a high pressure in the upper pouch which increases the risk of staple-line disruption. PMID- 9731678 TI - Changes in esophageal function after vertical banded gastroplasty as demonstrated by esophageal scintigraphy. AB - BACKGROUND: The effects of surgery for morbid obesity on the function of the upper gastrointestinal (GI) tract are of interest to bariatric surgeons. This study was undertaken to determine any changes in esophageal function, following vertical banded gastroplasty (VBG) in morbidly obese patients, as detected by esophageal scintigraphy. METHODS: Ten consecutive morbidly obese patients (six female and four male) underwent preoperative esophageal scintigraphy and upper GI endoscopy. These investigations were repeated 12 months after VBG to coincide with expected appreciable weight reduction. The results were tabulated together with body mass indices, crude weights and percentage excess weight lost. RESULTS: Before VBG one patient gave a history of mild heartburn, one had mild dyspepsia and the remaining eight patients had no GI symptoms. No patient had a hiatus hernia or endoscopic evidence of reflux esophagitis. Preoperatively all patients had abnormal scintiscans. The abnormalities were esophageal retention (all) and intraesophageal reflux (five out of 10 patients). Gastroesophageal reflux was not identified in any patient. Postoperatively scintiscans were normal or improved in six out of 10 patients and unchanged in four out of 10 patients. In three patients the scans were normal and three showed overall improvement in esophageal function, although in one of these latter patients gastroesophageal reflux was observed. CONCLUSIONS: In this series of morbidly obese patients, esophageal function as assessed by scintigraphy was abnormal. Following VBG it improved in six out of 10 patients and was unchanged in four out of 10. However, in one patient, who had shown an overall improvement in esophageal function, gastroesophageal reflux was demonstrated when it had not been seen preoperatively. This was asymptomatic. Thus, adverse changes in esophageal function after VBG were uncommon. PMID- 9731680 TI - Changes in measured resting energy expenditure after Roux-en-Y gastric bypass for clinically severe obesity are not related to bypass limb-length. AB - BACKGROUND: Roux-en-Y gastric bypass (RYGB) for clinically severe obesity (CSO) results in a 'paradoxical' response of the measured resting energy expenditure (MREE) in which the MREE remains within the predicted range based upon the Harris Benedict (HB) equation, despite a significant decrease in caloric intake to 500 1000 kcal/day. The mechanism for this response is unknown. A study was undertaken to determine whether the changes in MREE after RYGB are related to limb-length of the gastric bypass. METHODS: A prospective clinical trial of varying limb-lengths based on body mass index (BMI) in patients having RYGB for CSO. The records of patients who underwent RYGB for CSO and had MREE measured at baseline, 6 months and 12 months postoperation were reviewed. MREE was performed using a Med Graphics CCM system after an overnight fast or at least 4 hours after a light meal, and a 30 minute rest in a supine position in a neutral environment, on the same day of the week between the hours of 10 a.m. and 4 p.m. Patients were selected for RYGB in accordance with NIH recommendations. RYGB was performed in a standardized fashion with the Roux limb-length varied as follows: (A) BMI < or = 51 kg/m2 - 75 cm limb (n = 20); (B) BMI < or = 51 kg/m2 - 150 cm limb (n = 16); (C) BMI > or = 51 kg/m2 - 150 cm limb (n = 18); or (D) BMI > or = 51 kg/m2 - 250 cm limb (n = 6). RESULTS: Data from 60 patients (nine male, 51 female; mean age 39 years; mean baseline BMI 51.5 +/- 10 kg/m2; mean baseline weight 145 +/- 32 kg) were analyzed. There were no significant differences in MREE or percentage HB predicted energy expenditure between the groups. CONCLUSIONS: These data suggest that the observed changes in MREE following RYGB for CSO are not related to the limb-length of the bypass. PMID- 9731681 TI - Pitfalls in the diagnosis of gallbladder disease in clinically severe obesity. AB - BACKGROUND: Cholelithiasis affects 10-20% of the USA population, with higher incidence in certain ethnic groups. Obesity is associated with an increase in gallstone formation, reported in up to 45% of morbidly obese patients. Ultrasound is the best diagnostic tool, although its accuracy is less in this particular population. This paper discusses false negative sonographic findings in morbid obesity. METHODS: Retrospective review of 5257 patients submitted to bariatric surgery. Cholecystectomy had previously been performed in 16%. Gallbladder ultrasound was obtained in the remaining group, and cholecystectomy was done based on this information and/or intraoperative observations. Radiology results and surgical findings were correlated with pathology reports. Misread films were reviewed by a radiologist blind to these reports. RESULTS: The series consisted of 88% females. Mean age, weight and percentage overweight were 37 years, 125 kg and 105%, respectively. Cholecystectomy was performed in 3084 patients (59%). Discrepancies between radiological and pathological findings were found in 35 cases (1.1%). Five correct diagnosis of lithiasis also had gallbladder hydrops. Four 'inconclusive' and 20 'negative' studies showed definitive pathology. In six cases of 'non/poor visualization', lithiasis was encountered. CONCLUSIONS: Preoperative gallbladder ultrasound is mandatory in bariatric surgery. Results are accurate and false-negative reports rare if sonographers and radiologists are experienced. Non/poor visualization is usually due to technical problems or gallbladder pathology, not due to the patient's size. False-negative results are commonly caused by soft stones, microlithiasis or polypoid cholesterolosis. Single calculus impacted in the cystic duct can produce hydrops, resulting in a negative sonogram. PMID- 9731682 TI - Influence of a forced air warming system on morbidly obese patients undergoing Roux-en-Y gastric bypass. AB - BACKGROUND: Hypothermia during and after major abdominal surgery decreases host defenses, increases the incidence of coagulopathy and may alter blood pressure, cardiac contractility and myocardial stability. METHODS: We designed a prospective randomized study to compare the benefits of a forced air warming system with warm blanket treatments in minimizing the effects of hypothermia on 64 morbidly obese patients undergoing Roux-en-Y gastric bypass. RESULTS: Patients in the forced air warming group (n = 32) had significantly higher perioperative body core temperature, lower central venous pressure and blood pressure readings, lower incidence of shivering, less blood loss intraoperatively and achieved a higher post anesthesia Aldrete Score than those patients in the warmed blanket group (n = 32). CONCLUSION: The forced air warming system is safe, cost effective and beneficial in minimizing the undesirable consequences of hypothermia in morbidly obese patients undergoing Roux-en-Y gastric bypass. PMID- 9731683 TI - Pregnancy following gastric bypass for morbid obesity. AB - BACKGROUND: Women who suffer from morbid obesity are often infertile. If these women are able to become pregnant, they are considered high risk because of the hypertension, diabetes and other associated risk factors. Following the pregnancy is difficult due to limitations of the physical examinations. More costly ultrasound examinations are needed at a higher frequency. Bariatric surgery reduces the woman's weight and the incidence of obesity related co-morbidities. The number of pregnancies and rate of complications during those pregnancies in our post-bariatirc surgical patients were evaluated. METHOD: Our group has been doing bariatric surgery since the early 1980s. We have over 2000 active patients on our current newsletter mailing list. The patients also have a series of networks through support groups. The patients are informed to contact us when they become pregnant so we may assist the obstetrician with their care. Through these various means, we have been able to identify 41 women in our patient population who have become pregnant. Using personal interview, questionnaire, and review of perinatal records, pregnancy-related risks and complications were studied. RESULTS: With over a 95% follow-up rate on the patients identified as having been pregnant following surgery, we found less risk of gestational diabetes, macrosomia, and cesarean section than associated with obesity. There were no patients with clinically significant anemia. CONCLUSION: Since the patients had an operation that restricts their food intake, some basic precautions should be taken when they become pregnant. With this in mind, our patients have done well with their pregnancies. The post-surgical group had fewer pregnancy-related complications than did an internally controlled group that were morbidly obese during their previous pregnancies. PMID- 9731684 TI - Silicone-adjustable gastric banding: disappointing results. AB - BACKGROUND: Silicone-adjustable Gastric banding (SAGB) has been popularized as a minimally invasive, completely reversible surgical treatment for morbid obesity. We report here our 3-year experience of SAGB with special reference to complications and side-effects. METHODS: There were 90 patients in total, of whom 72 were women. Median age was 42 (range, 20-68) years and median body mass index (BMI) was 43 (range, 34-57) kg/m2. Laparoscopy was attempted to position the band in 63 cases but had to be converted to laparotomy in 16 (25%). Twenty-seven patients were laparotomized. We used the Swedish band (AB Obtech) throughout the series. In addition to regular clinic visits, patients were followed-up with upper gastrointestinal series 6 months postoperatively and gastroscopy after 2 years or earlier when symptomatic. RESULTS: Median BMI decreased to 32 kg/m2 after 12 months and to 31 kg/m2 after 24 months. With a median follow-up time of 35 months (range, 22-48), 32 patients (35%) have been re-operated usually with removal of the balloon system and conversion into a Roux-en-Y gastric bypass. The most common reasons for re-operation were band erosion (n = 10) and erosive esophagitis (n = 14). Additional indications for re-operation included pouch dilatation, invagination of distal gastric wall through the band, leakage from the balloon, patient dissatisfaction, and severe allergic reaction. When questioned 2 years postoperatively more than half of the patients reported vomiting, heartburn and regurgitation but 78% still pronounced themselves satisfied with the operation. Esophagitis was found in 56% of the patients at gastroscopy after 2 years. CONCLUSION: SAGB could be positioned with laparoscopy in 75% of the cases but the incidence of complications and side-effects postoperatively has been high. PMID- 9731685 TI - Double closed loop obstruction and perforation in a previous Roux-en-Y gastric bypass. AB - A healthy 45-year-old woman with a previous Roux-en-Y gastric bypass presented with the signs, symptoms and blood analysis results consistent with acute pancreatitis. She was initially treated nonoperatively and subsequently went into circulatory shock. Computerized tomographic scan and exploratory laparotomy revealed a volvulus of the afferent jejunal limb with secondary obstruction, necrosis, and perforation of the bypassed stomach. PMID- 9731686 TI - Obesity classification. PMID- 9731687 TI - On reporting variations on vertical banded gastroplasty. PMID- 9731688 TI - Current insights and new treatment options in heart failure. PMID- 9731689 TI - Left ventricular dilatation after myocardial infarction: ACE inhibitors, beta blockers, or both? AB - Left ventricular (LV) dilatation after myocardial infarction (MI) is a major predictor of prognosis and identifies which patients will develop heart failure. Left ventricular dilatation or remodeling starts immediately after MI and progresses in the chronic phase of heart failure. Factors influencing remodeling, such as infarct size and neurohumoral activation, including the sympathetic and renin-angiotensin system, are discussed. Remodeling can be affected by reduction of infarct size and inhibition of neurohumoral activation. The effect of thrombolysis, beta-blockade, and angiotensin-converting enzyme (ACE) inhibition in the acute phase after MI and in the chronic phase of heart failure on remodeling are discussed. On the basis of beneficial effects of ACE inhibition and beta-blockade in acute MI and in chronic heart failure, a treatment strategy is proposed in which both ACE inhibition and beta-blockade are started early after MI. Depending on infarct size and ventricular function, continued treatment in the chronic phase of heart failure must be considered. PMID- 9731690 TI - Hypertension and the development of heart failure. AB - Hypertension is a well-known risk factor that predisposes to the development of left ventricular hypertrophy, coronary flow abnormalities, and systolic and diastolic dysfunction. This complex of abnormalities is known as hypertensive heart disease and eventually leads to heart failure. Structural lesions underlying this process include excess deposition of collagens and cellular infiltration, with an increase in the size of cardiac myocytes (remodeling). The occurrence of arteriolar lesions may lead to impediments of flow. Clinically, hypertensive heart disease manifests itself by anginal complaints and sometimes by silent ischemia, arrhythmias, and sudden death. Alterations in systolic and/or diastolic function may be associated with symptoms of heart failure. Although successful treatment of hypertension can probably retard the process of cardiac impairment, there is as yet no evidence that heart failure can be prevented. PMID- 9731691 TI - Neurohumoral activation and progression of heart failure: hypothetical and clinical considerations. AB - The model for heart failure has changed radically over the past 20 years. No longer a simple hemodynamic paradigm of pump dysfunction, heart failure is now characterized as a complex clinical syndrome with release of many neurohormones and cytokines, which are believed to be most responsible for progression of disease. This change in our understanding of the pathophysiology of heart failure has important therapeutic implications. Drugs designed to influence the myocardial contractile state have been found over the past few decades to have either a neutral or an adverse effect on long-term survival, whereas agents designed to block the renin-angiotensin-aldosterone and other neurohormonal systems have proved to be remarkably effective treatment. Recently, drugs designed to block excessive sympathetic nervous system activity have been demonstrated in well-controlled studies to be safe and effective forms of therapy for heart failure. Carvedilol, a nonselective beta-adrenergic blocker with alpha1 blocking and antioxidant properties, is associated with prevention of progression of heart failure as manifested by improvement in left ventricular (LV) function, reduction in heart size, and improved survival in patients with New York Heart Association functional Class II and III symptoms. This improvement is observed equally in patients with ischemic and non-ischemic heart failure. It is tempting to speculate that beta-adrenergic blockers prevent the progression of heart failure by reducing LV mass and LV chamber size. In essence, carvedilol, and perhaps other beta-adrenergic blockers, appear to abrogate relentless LV remodeling which is typically associated with progression of heart failure. The combination of angiotensin-converting enzyme inhibitors and beta-adrenergic blockers may be particularly effective in this regard, although more data on beta adrenergic blockers in patients with advanced heart failure are needed. Data from experimental heart failure animal models also suggest that endothelin (ET) subtype A (ET(A)) receptor blockers have the potential to lessen the pace of progressive LV remodeling. As our understanding of the neuroendocrine response to diminished cardiac performance improves, novel and even more imaginative neurohormonal and cytokine antagonists are likely to emerge as important new treatments for both hypertension and heart failure. PMID- 9731692 TI - Neurohormonal activation, oxygen free radicals, and apoptosis in the pathogenesis of congestive heart failure. AB - A variety of pathophysiologic processes are activated in patients with congestive heart failure (CHF), and some of these have been implicated in the progression of the disease. The most important processes to be activated in CHF are the neurohormonal systems, which include the renin-angiotensin system, the sympathetic nervous system, and the endothelin system. In addition to the neurohormonal systems, the formation of reactive oxygen free radicals is increased in patients with CHF. It has been postulated that stimulation of neurohormonal pathways and the formation of oxygen free radicals ultimately lead to the activation of a family of transcription factors that are involved in cardiac remodeling, which is a hallmark of CHF. In addition, the formation of oxygen free radicals has been implicated in the process of apoptosis or programmed cell death, which may be responsible for a continued loss of myocardial cells, resulting in the progressive decrease in left ventricular function that occurs over time in patients with CHF. Carvedilol is a multiple action neurohormonal antagonist that is effective in slowing the progression of CHF. In double-blind, placebo-controlled clinical trials, carvedilol decreased mortality by 65% (p <0.001) and significantly reduced hospitalization. Carvedilol is a nonselective beta-blocker and vasodilator, the latter activity resulting from alpha1-adrenoceptor blockade. The hemodynamic responses produced by carvedilol result primarily from the blockade of beta1-, beta2-, and alpha1 adrenoceptors. Carvedilol reduces total peripheral vascular resistance and preload without significantly compromising cardiac output or eliciting reflex tachycardia. Carvedilol is also a potent antioxidant that may protect the myocardium from damage produced by oxygen radicals and, as a consequence of its antioxidant activity, carvedilol also inhibits apoptosis in the myocardium. The ability of carvedilol to inhibit apoptosis in the heart may be responsible, in part, for the ability of the drug to reduce mortality and to inhibit the progression of CHF. PMID- 9731693 TI - Ejection fraction improvement by beta-blocker treatment in patients with heart failure: an analysis of studies published in the literature. AB - Because ejection fraction (EF) is one of the most important predictors of survival in patients with left ventricular (LV) dysfunction and because Packer showed a large reduction in mortality figures with carvedilol, in contrast to former studies with bisoprolol and metoprolol, we investigated if this difference in survival may be related to a difference in improvement of LV function by different beta-blockers. We searched the MEDLINE database and all reference lists of articles obtained through the search for the relation between beta-blocker treatment and improvement in EF. Forty-one studies met the criteria and we added two of our own studies. Four hundred and fifty-eight patients were treated with metoprolol with a mean follow-up of 9.5 months and a mean increase in EF of 7.4 EF units. One thousand thirty patients were treated with carvedilol with a mean follow up of 7 months and a mean increase in EF of 5.7 EF units. One hundred ninety-nine patients were treated with bucindolol with a mean follow-up of 4 months and a mean increase in EF of 4.6 EF units. Several small studies with nebivolol, atenolol, and propranolol were also studied and, when combined, the mean increase in EF was 8.6 EF units. When patients with idiopathic and ischemic cardiomyopathies were compared, the average increase in EF units was 8.5 vs. 6.0, respectively. The use of beta-blocker treatment in heart failure patients, irrespective of the etiology, improved LV function in almost all studies and it appears that the differences among beta-blockers and among etiologies is small and probably insignificant. However, there is a difference in survival rate when the various beta-blockers are compared, suggesting that mechanisms other than improvement of LV function by beta-blockers are responsible for the difference in survival. PMID- 9731694 TI - Maximal and submaximal exercise testing in heart failure. AB - Although reduced exercise capacity is the main complaint of patients with congestive heart failure (CHF), the best method to measure it remains controversial. Peak VO2, obtained using maximal exercise testing, is the most accurate measure of maximal functional capacity. It is related to peak exercise cardiac output and is one of the most important independent variables for the prognostic assessment of patients with CHF. It has, however, a low sensitivity for measurement of changes induced by therapy and is poorly related to everyday physical activity, patient symptoms, and quality of life. The anerobic threshold may also be regarded as a parameter of maximal functional capacity. Its value is mainly indirect, because it shows that the patient is performing a maximal effort limited by the cardiovascular system. The VO2 kinetics at the start and at the end of exercise are probably more related to patient symptoms, but it is unresolved which protocols and parameters might best be used to study this aspect of exercise performance. Duration of a submaximal exercise at a constant work rate and the distance walked during a 6-min walking test are gaining wide popularity as parameters of submaximal performance. However, when these exams are carried out up to exhaustion in patients with severe functional limitation, they may involve attainment of the anerobic threshold and therefore their clinical meaning may be similar to the one of a maximal exercise test. Moreover, tests based on the assessment of submaximal exercise capacity have been useful for assessment of therapy in single-center trials but have been often inadequate in multicenter trials. PMID- 9731696 TI - Beta-blockers for chronic heart failure: from prejudice to enlightenment. AB - Experience accumulated from several large trials strongly suggest that beta blockers should be used for the management of congestive heart failure (CHF). Beta-blockade should be added to conventional therapy such as diuretics, ACE inhibitors, and digoxin, as this was the approach used in the major trials. It is appropriate to treat patients with mild, moderate and, when stable, severe CHF. The benefits obtained include improvements in left ventricular function, reductions in symptoms and morbidity, improvement of quality of life, and delay of clinical progression, reflected by a reduced need for hospitalization and a reduction in mortality. Beta-blockers are much better tolerated, when used appropriately in selected patients, than was previously supposed. PMID- 9731695 TI - Ischemia and left ventricular dysfunction: a reciprocal relation? AB - There is convincing evidence that (prolonged) episodes of myocardial ischemia lead to impairment of left ventricular (LV) function and ultimately to chronic congestive heart failure (CHF), but whether the opposite is also true has not been well established. We studied this issue in two groups of CHF patients with positron emission tomography (PET) by using [13N]ammonia (13NH3) as a tracer. In the first protocol we compared 12 patients with idiopathic dilated cardiomyopathy (who have normal coronary arteries) with 12 healthy controls. In the second protocol we studied a group of 24 patients with documented coronary artery disease (CAD). In this protocol, we compared patients with normal LV function to those with LV dysfunction and CHF. In patients with cardiomyopathy, myocardial blood flow at rest was normal but flow reserve (after dipyridamole infusion) was significantly impaired (1.7 +/- 0.08) compared with normal subjects (2.7 +/- 0.04; p <0.05). Furthermore, by examining [18F]fluorodeoxyglucose (18FDG) uptake, a perfusion-metabolism mismatch was observed in 24 +/- 6% of the myocardium in patients with cardiomyopathy as opposed to 0% of normals (p <0.05). In patients with CAD, myocardial blood flow reserve (measured in non-stenotic arteries to non infarcted area) was impaired in CHF patients (1.7 +/- 0.06) compared to those with normal LV function (2.3 +/- 0.05; p <0.05). In both groups of CHF patients, the impairment of blood flow reserve showed a significant correlation with the severity of CHF. In conclusion, myocardial blood flow reserve is impaired in patients with CHF in proportion to the degree of CHF. Metabolic studies with 18FDG further show that, in patients with idiopathic dilated cardiomyopathy and CHF, flow-metabolism mismatch is present in a substantial part of the myocardium, suggesting a pathogenetic role for ischemia. PMID- 9731697 TI - Retinoid X receptor and c-cerbA/thyroid hormone receptor regulate erythroid cell growth and differentiation. AB - Nuclear receptors are important regulators of erythroid cell development. Here we investigated the impact of retinoid X receptor (RXR), retinoic acid receptor (RAR), and of the c-erbA/thyroid hormone (T3) receptor (c-erbA/TR) on growth and differentiation of erythroid cells using an in vitro culture system of stem cell factor-dependent erythroid progenitors. RXR, RAR, and c-erbA/TR-specific ligands were found to induce erythroid-specific gene expression and to accelerate erythroid differentiation in culture, with T3 being most effective. Furthermore, while ligand-activated c-erbA/TR accelerated differentiation, unliganded c erbA/TR effectively blocked differentiation and supported sustained progenitor growth in culture. Thus, c-erbA/TR appears to act as a binary switch affecting erythroid cell fate: unliganded c-erbA/TR supports growth while ligand-activated c-erbA/TR induces differentiation. Additionally, to determine the impact of RXR for erythroid cell development, dominant interfering mutant RXRs, lacking the transcriptional activator functions AF-1 and AF-2, or AF-2 only, or the entire DNA-binding domain, were introduced into erythroid progenitor cells via recombinant retrovirus vectors and analyzed for RXR-specific effects. It was found that expression of wild-type RXR and of the RXR mutants devoid of AF-1 and/or AF-2 supported a transient outgrowth of erythroid cells. In marked contrast, expression of the dominant interfering deltaDNA-binding domain RXR, containing a deletion of the entire DNA-binding domain, was incompatible with erythroid cell growth in vitro, suggesting a pivotal role of RXR for erythroid cell development. PMID- 9731698 TI - Differential regulation of basal and cyclic adenosine 3',5'-monophosphate-induced somatostatin gene transcription in neural cells by DNA control elements that bind homeodomain proteins. AB - A number of genes encoding neuropeptides are expressed in the peripheral and central nervous systems, in different endocrine organs, and in specialized cells distributed along the gastrointestinal tract. Whether expression of the same neuropeptide gene in different tissues is regulated by similar transcriptional mechanisms or by mechanisms that differ in a cell-specific manner remains unclear. We report on promoter studies on the regulation of the somatostatin gene in immortalized neural precursor cells derived from developing rat forebrain. Expression of the somatostatin gene in these cells was determined by RT PCR/Southern blot analysis, by immunocytochemistry, and by RIA. We show that in cerebrocortical and hippocampal cells, expression of the somatostatin gene is regulated by several negative and positive DNA cis-regulatory elements located throughout the promoter region. The somatostatin cAMP-response element appears to play a prominent role in neural somatostatin gene expression by acting as a strong enhancer even in the absence of cAMP stimulation. Site-directed mutagenesis followed by transient transfection assays indicated that SMS-TAAT1, SMS-TAAT2, and SMS-UE, three previously identified homeodomain protein-binding regulatory elements that enhance transcription in pancreatic cells, act as repressors of transcription in neural cells. Electrophoretic mobility shifts assays indicate that those elements bind protein complexes that differ between neural and pancreatic cells. Our results support the notion that expression of the somatostatin gene in neural cells occurs via transcriptional mechanisms that are different from those regulating expression of the same gene in pancreatic cells. PMID- 9731700 TI - Upstream stimulatory factors mediate estrogen receptor activation of the cathepsin D promoter. AB - Overexpression of cathepsin D (CD), a ubiquitous lysosomal protease, is closely associated with a poor clinical outcome for patients with breast cancer. Estrogen greatly induces transcription of the CD gene in estrogen receptor (ER)-positive breast cancer cells. In this report, we transiently introduced a human CD promoter/chloramphenicol acetyltransferase reporter gene into human MCF-7 breast cancer cells to study the mechanisms by which the ER activates the promoter. Using an in vivo Exonuclease III footprinting assay, we found that estrogen stimulation of MCF-7 cells induced loading of a transcription factor(s) to a portion of the promoter (-124 to -104) that is homologous to the adenovirus major late promoter element. Subsequent gel mobility shift assays with a 21-bp CD -124/ 104 probe and nuclear extracts prepared from naive and estrogen-stimulated cells detected a single sequence-specific protein-DNA complex. Southwestern and UV cross-linking experiments detected two proteins of 44 kDa and 43 kDa that were specifically bound to the 21-bp fragment of the promoter. Gel super-shift assays with upstream stimulatory factor 1 (USF-1) and USF-2 antibodies demonstrated that USF-1 and USF-2 bound to the E box probe. Sequence specific binding was abolished by a 2-bp change shown previously to prevent the binding of USF to the E box. Incorporation of a mutant E box into the wild-type CD promoter/chloramphenicol acetyltransferase gene abolished USF binding and reduced the levels of both basal and estrogen-stimulated transcription. These results suggest that the ER targeting of USF-1 and USF-2 is a critical step in hormone activation of CD gene transcription in human breast cancer cells. PMID- 9731699 TI - IRE-ABP (insulin response element-A binding protein), an SRY-like protein, inhibits C/EBPalpha (CCAAT/enhancer-binding protein alpha)-stimulated expression of the sex-specific cytochrome P450 2C12 gene. AB - In primary hepatocytes, overexpression of an insulin response element-A binding protein (IRE-ABP), a member of the SRY family of high-mobility group (HMG) proteins, inhibits CCAAT/enhancer-binding protein alpha (C/EBPalpha)-mediated activation of the female-specific cytochrome P450 2C12 (CYP2C12) gene, but not the male-specific cytochrome P450 2C11 (CYP2C11) gene. IRE-ABP and C/EBPalpha have overlapping specificity for the C/EBPalpha target site in the CYP2C12 promoter and compete for binding to CYP2C12 DNA in vitro. In contrast, IRE-ABP and C/EBPalpha bind distinct sequences in the CYP2C11 promoter. A single amino acid substitution in the HMG domain of IRE-ABP impairs its ability to bind DNA and to inhibit the effect of C/EBPalpha on CYP2C12 gene expression. Therefore, the ability of IRE-ABP to inhibit C/EBPalpha-stimulated CYP2C12 gene expression requires a functional DNA-binding domain. Taken together, our findings suggest that SRY-like proteins can bind to a subset of sequences recognized by the C/EBP family of DNA-binding proteins and modulate gene transcription in a context specific manner. PMID- 9731701 TI - Transrepression of c-jun gene expression by the glucocorticoid receptor requires both AP-1 sites in the c-jun promoter. AB - The c-jun protooncogene encodes a nuclear protein, cJun, which is a major component of the AP-1 transcription factor. AP-1 regulates various aspects of cell proliferation and differentiation. As an immediate early response gene, the expression of the c-jun gene is affected by various extracellular stimuli, such as serum, phorbol esters, and glucocorticoids. In mouse L929 fibroblasts, dexamethasone (DEX) treatment caused a 60% reduction of c-jun mRNA levels. Previous studies indicated that this reduction is due to the alteration of the transcription rate of the c-jun gene. To further investigate the molecular mechanisms of transcriptional repression of c-jun by DEX, a full-length human c jun promoter, from -1780 to +731, was amplified from genomic DNA using PCR and then linked to the luciferase reporter gene. To identify the regulatory elements responsible for the down-regulation, nested deletions spanning the promoter were generated, and the promoter/luciferase constructs were transiently transfected into L929 cells. Upon hormone treatment, basal activity of the full-length c-jun promoter was reduced by approximately 40%, which accounts for two-thirds of the overall down-regulation observed at the mRNA level. This reduction of c-jun promoter activity was abolished after deletion of the region between -1780 to 63, where two AP-1 sites (-182 and -64) are located. Site-directed deletion of these AP-1 sites reduced the basal activity of the c-jun promoter and prevented repression by DEX. Repression of the c-jun gene is due to the transrepression activity of the glucocorticoid receptor (GR), as determined using GR mutants lacking this activity. Overexpression of cJun overcame the negative effect of DEX, suggesting that down-regulation of the c-jun gene by hormone is mediated by the interaction between the GR and the cJun protein. These studies are the first to show that glucocorticoids can repress c-jun promoter activity through the AP-1 sites in the c-jun promoter in mouse fibroblast cells. They also suggest that inhibition of cell proliferation by glucocorticoids may be due not only to the interference with AP-1 activity on other cellular genes, but also because of a direct transcriptional suppression of c-jun gene expression by the GR. PMID- 9731702 TI - Definition of a negative modulation domain in the human progesterone receptor. AB - The progesterone receptor (PR) occurs in two major forms, the full-length PRB and the amino-truncated PRA, which lacks 164 amino-terminal residues. PRB functions as a strong transcriptional activator of progesterone-responsive genes, whereas PRA is inactive in several cell types where it may even act as a trans-dominant repressor of PRB and other steroid receptors, like the glucocorticoid receptor or, reportedly, the estrogen receptor. We initially observed that a PR deleted of its entire amino domain (PR538-C) is incapable of trans-repressing PRB or glucocorticoid receptor, suggesting that a negative modulation domain must be contained in the region between position 165 and 538. After testing progressive deletion mutants and chimeras, we demonstrate that this negative modulating domain is confined within 120 residues in the amino-terminal region and that it contains a subdomain of 40 residues that is crucial for intermolecular transrepression. Duplication, deletion, and transplantation of the negative modulation domain show that the negative modulation domain has only a limited functional autonomy. In our hands, transrepression of estrogen receptor could not be substantiated, and, under our conditions, at least an equimolar concentration of PRA expression plasmid is required for transrepression. Our deletion studies reveal domains that correlate with strong homology patches between the amino terminal domains of mammalian and avian PR. PMID- 9731703 TI - Inhibition by insulin of glucocorticoid-induced gene transcription: involvement of the ligand-binding domain of the glucocorticoid receptor and independence from the phosphatidylinositol 3-kinase and mitogen-activated protein kinase pathways. AB - Insulin can inhibit the stimulatory effect of glucocorticoid hormones on the transcription of genes coding for enzymes involved in glucose metabolism. We reported earlier that insulin inhibits the glucocorticoid-stimulated transcription of the gene coding for liver 6-phosphofructo-2-kinase (PFK-2). To elucidate the mechanism of these hormonal effects, we have studied the regulatory regions of the PFK-2 gene in transfection experiments. We found that both glucocorticoids and insulin act via the glucocorticoid response unit (GRU) located in the first intron. Footprinting experiments showed that the GRU binds not only the glucocorticoid receptor (GR), but also ubiquitous [nuclear factor I (NF-I)] and liver-enriched [hepatocyte nuclear factor (HNF)-3, HNF-6, CAAT/enhancer binding protein (C/EBP)] transcription factors. Site-directed mutational analysis of the GRU revealed that these factors modulate glucocorticoid action but that none of them seems to be individually involved in the inhibitory effect of insulin. We did not find an insulin response element in the GRU, but we showed that insulin targets the GR. Insulin-induced inhibition of the glucocorticoid stimulation required the ligand-binding domain of the GR. Finally, the insulin-signaling cascade involved was independent of the phosphatidylinositol-3-kinase and mitogen-activated protein kinase pathways. Together, these results suggest that insulin acts on the PFK-2 gene via another pathway and targets either the GR in its ligand-binding domain or a cofactor interacting with this domain. PMID- 9731704 TI - A tumor-specific truncated estrogen receptor splice variant enhances estrogen stimulated gene expression. AB - This study examines the cooperative effects of a human estrogen receptor-alpha (ERalpha) isoform on estrogen (E2)-mediated gene activation in U2-OS osteosarcoma cells. Delta5ERalpha, an alternatively spliced ERalpha variant lacking exon 5, is coexpressed with normal ERalpha in several E2-responsive neoplastic tissues. However, the potential interactions of delta5ERalpha with normal ERalpha have not been functionally characterized. Delta5ERalpha encodes the hormone-independent trans-activating function (AF-1), as well as the constitutive receptor dimerization and DNA-binding domains. It is generated by an alternate splice event that omits exon 5 and alters the reading frame of the resulting mRNA. The delta5ERalpha protein is prematurely truncated and lacks the majority of the hormone-binding and activating function-2 (AF-2) domains. When delta5ERalpha mammalian expression vector was transfected alone in human ERalpha/ERbeta negative osteosarcoma U2-OS cells, it had no effect on either basal or E2 mediated EREtk81Luc reporter transcriptional activity, while transfected cells expressing control normal ERalpha increased EREtk81 Luc activity up to 20-fold in response to 10 nM E2. However, when delta5ERalpha was cotransfected with normal ERalpha, both basal and E2-stimulated EREtk81Luc reporter activation were increased approximately 500% over levels observed when cells were transfected with ERalpha alone. Similar effects of delta5ERalpha and normal ERa coexpression were observed using an E2-responsive human C3 promoter/luciferase reporter construct. The effects of delta5ERalpha on normal ERalpha were further assessed in U2-OS cells stably transfected with normal ERalpha. Transfection of increasing amounts of delta5ERalpha expression vector into [ERalpha+]OS cells resulted in potentiation of E2-stimulated ERELuc activity in a synergistic, dose-dependent manner. Moreover, coexpression of delta5ERalpha in [ERalpha+]OS cells improved E2 sensitivity 100-fold over cells expressing ERalpha alone. Proliferation rates of stable U2-OS cell lines expressing delta5ERalpha were significantly increased (P < 0.05), with cell doubling times reduced from 35 h in control parental U2-OS cells to 28 h in [delta5ERalpha]OS cells. However, growth rates were not affected by either E2 or tamoxifen treatment. Electromobility shift/supershift assays using nuclear extracts of U2-OS cells stably transfected with ERalpha and delta5ERalpha confirmed the constitutive binding of delta5ERalpha and ERalpha protein to estrogen-response element (ERE) sequence independent of E2 and also showed an increase in delta5ERalpha/ERalpha-ERE complexes with E2 treatment. These data are consistent with interactive effects of normal ERalpha and delta5ERalpha on transcription from classic ERE gene promoters. Delta5ERalpha appears to therefore act as a dominant positive receptor that increases both basal and E2-stimulated gene transactivation of normal ERalpha. PMID- 9731705 TI - Thyroid hormone receptor does not heterodimerize with the vitamin D receptor but represses vitamin D receptor-mediated transactivation. AB - The 9,000 Mr calcium-binding protein calbindin-D9k (CaBP9k) is markedly induced by 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] in mammalian intestine. However, although a vitamin D response element (VDRE) has been reported in the promoter of the rat CaBP9k gene (at -490/-472), the CaBP9k promoter is weakly transactivated by 1,25-(OH)2D3. Previous studies indicated that when MCF-7 cells are transfected with the rat CaBP9k VDRE ligated to the thymidine kinase promoter and treated with both 1,25-(OH)2D3 and T3 there is an enhancement of the response observed with 1,25-(OH)2D3 alone, suggesting direct cross-talk between thyroid hormone and the vitamin D endocrine system and activation via the formation of vitamin D receptor (VDR)-thyroid hormone receptor (TR) heterodimers. To determine whether the weak response of the rat CaBP9k natural promoter to 1,25-(OH)2D3 could be enhanced by T3, CaBP9k promoter/reporter chloramphenicol acetyltransferase constructs were transfected in MCF-7 cells, and the cells were treated with the two hormones alone or in combination. No induction with T3 alone and no enhancement of reporter activity in the presence of both hormones was observed. To determine whether a lack of effect by T3 was specific for the CaBP9k promoter and to further examine the possibility of cross-talk between the TR- and VDR signaling pathways, the 1,25-(OH)2D3-responsive rat 24 hydroxylase [24(OH)ase] promoter and the rat osteocalcin VDRE (-457/-430), both fused to reporter genes were similarly examined in MCF-7 cells. Again, no enhancement of the response to 1,25-(OH)2D3 was observed in the presence of T3. In addition, a similar lack of response to T3 but responsiveness to 1,25-(OH)2D3 was observed when UMR106-01 osteosarcoma cells [which, like MCF-7 cells, express VDR, TR, and the retinoid X receptor (RXR) endogenously] were transfected with a 1,25-(OH)2D3 responsive mouse osteopontin promoter reporter. In vitro DNA binding assays were carried out using purified human VDR, human RXRalpha, and chick T3Ralpha and 24(OH)ase, osteocalcin, osteopontin, and CaBP9k VDRE oligonucleotide probes. No VDR-TR heterodimer binding on any of these VDREs was observed, although, as expected, there was binding by the VDR-RXR complex and strong TR-RXR binding to a consensus thyroid hormone response element. Simultaneous gel retardation assays using similar and lower concentrations of TR with RXR showed strong binding of TR-RXR on a 32P-labeled thyroid response element. Studies using the yeast two-hybrid system also did not provide evidence for the formation of a VDR-TR protein protein interaction. In addition, in vivo data showed that transfection of TR, in fact, repressed VDR-mediated transcription and that the repression could be reversed by the addition of RXR. Thus, in vitro and in vivo experiments do not support ligand-sensitive transactivation mediated by VDR-TR heterodimer formation but rather suggest that TR expression can repress 1,25-(OH)2D3-induced transcription predominantly by sequestering RXR. PMID- 9731706 TI - Characterization of the DNA-binding and dominant negative activity of v-erbA homodimers. AB - The oncoprotein v-erbA is a mutated form of thyroid hormone receptor alpha1 that is virtually incapable of binding T3. V-erbA is a dominant repressor of transcription induced by thyroid hormone receptors and retinoic acid receptors; however, the genetic targets of v-erbA that lead to oncogenesis are not known. Although v-erbA can bind as monomers and dimers to DNA containing the consensus sequence AGGTCA arranged as direct, inverted, or everted repeats, it is not known which sequence represents the optimal v-erbA-binding site. Determination of the DNA recognition properties of v-erbA would allow a better understanding of the repressor activity of this oncoprotein. The current studies, by using a random DNA selection strategy, have determined that the imperfect everted repeat 5' TGACC(T/C)NT(A/G)AGGTCAC is the optimal v-erbA homodimer-binding site, where N represents any di- or trinucleotide. Functional studies show that everted repeats containing this sequence are substantially more potent v-erbA response elements than direct or inverted repeats, even though many classic T3 response elements are direct repeats. Thus, v-erbA represses only a subset of T3 response elements. In a similar fashion, v-erbA was found to repress a subset of vitamin D response elements. Of general interest, the data indicate that the two molecules of a transcription factor homodimer do not necessarily have identical DNA-binding specificities. PMID- 9731707 TI - Stat 5b and the orphan nuclear receptors regulate expression of the alpha2 macroglobulin (alpha2M) gene in rat ovarian granulosa cells. AB - Alpha2-macroglobulin (alpha2M) is a serine protease inhibitor and cytokine inactivator associated with inflammation and tissue remodeling. The gene encoding this protein is selectively induced in the rat corpus luteum by the luteotropic hormone and cytokine, PRL. The promoter of the alpha2M gene contains two regulatory regions that bind a diverse set of transcription factors and confer functional activity in ovarian granulosa-luteal cells. The PRL response element (PRLRE) binds PRL-activated (tyrosine-phosphorylated) signal transducers and activators of transcription (Stat 5b and Stat 5a). 5'-Deletion of the Stat binding sites or mutation of either one or both of these sites within the context of the intact promoter abolished PRL inducibility of alpha2M promoter-reporter constructs in granulosa-luteal cells. Cotransfection with a vector expressing a dominant negative, truncated form of Stat 5b abolished PRL-induced activation of a2M transgenes. 5'-Deletion of the Stat-binding sites abolished all promoter reporter activity in response to PRL. Internal deletion of a second functional domain 3' of the PRLRE also abolished PRL inducibility and markedly reduced basal activity, indicating that functional interactions between these two regions might occur. The 3'-region was shown to bind orphan members of the nuclear receptor superfamily, steroidogenic factor 1 (SF-1) and chicken ovalbumin upstream promoter-transcription factor (COUP-TF) and has been called the orphan receptor response element (ORRE). When site-specific mutations were made in either the SF 1 -binding site or the two COUP-TF direct repeat (DR1 and DR2) binding sites in the context of the intact promoter, specific changes in the functional activity of this novel region of the alpha2M promoter were observed. Mutation of the SF-1 site drastically reduced basal activity of the alpha2M promoter. Mutation of the COUP-TF sites caused the basal activity of the alpha2M promoter to increase markedly. Neither mutation altered the PRL inducibility of these constructs. Lastly, differentiation of cultured granulosa cells was required for functional activity of both the PRLRE and the ORRE. Collectively, these results document for the first time that Stat 5b, SF-1, and COUP-TF each exert specific effects on the function of the alpha2M promoter: basal activity is controlled by the balance of SF-1 (positive) and COUP-TF (negative) activities and PRL inducibility is mediated by activation of Stat 5b. These results add alpha2M to the list of nonsteroidal genes regulated by SF-1 in the gonads and provide the first evidence that COUP-TF has a specific role in regulating ovarian gene activity. In addition, the ORRE and PRLRE act independently of, rather than synergistically with, each other to regulate basal and PRL-induced expression of alpha2M in ovarian luteal cells. PMID- 9731708 TI - Visualization of Pit-1 transcription factor interactions in the living cell nucleus by fluorescence resonance energy transfer microscopy. AB - The pituitary-specific transcription factor Pit-1 forms dimers when interacting with specific DNA elements and has been shown to associate with several other nuclear proteins. Recently, techniques have become available that allow visualization of protein-protein interactions as they occur in single living cells. In this study, the technique of fluorescence resonance energy transfer (FRET) microscopy was used to visualize the physical interactions of Pit-1 proteins fused to spectral variants of the jellyfish green fluorescent protein (GFP) that emit green or blue light [blue fluorescent protein (BFP)]. An optimized imaging system was used to discriminate fluorescence signals from single cells coexpressing the BFP- and GFP-fusion proteins, and the contribution of spectral overlap to background fluorescence detected in the FRET images was established. Energy transfer signals from living cells expressing a fusion protein in which GFP was tethered to BFP by short protein linker was used to demonstrate acquisition of FRET signals. Genetic vectors encoding GFP- and BFP Pit-1 proteins were prepared, and biological function of the fusion proteins was confirmed. FRET microscopy of HeLa cells coexpressing the GFP- and BFP-Pit-1 demonstrated energy transfer, which required the two fluorophores to be separated by less than 100 A. Biochemical studies previously demonstrated that Pit-1 physically interacts with both c-Ets-1 and the estrogen receptor. FRET imaging of cells coexpressing BFP-Pit-1 and GFP-Ets-1 demonstrated energy transfer between these fusion proteins, a result consistent with their association in the nucleus of these living cells. In contrast, there was no evidence for energy transfer between the BFP-Pit-1 and an estrogen receptor-GFP fusion proteins. It is likely that the FRET imaging approach described here can be applied to many different protein-partner pairs in a variety of cellular contexts. PMID- 9731709 TI - The DNA-binding and tau2 transactivation domains of the rat glucocorticoid receptor constitute a nuclear matrix-targeting signal. AB - Using an ATP-depletion paradigm to augment glucocorticoid receptor (GR) binding to the nuclear matrix, we have identified a minimal segment of the receptor that constitutes a nuclear matrix targeting signal (NMTS). While previous studies implicated a role for the receptor's DNA-binding domain in nuclear matrix targeting, we show here that this domain of rat GR is necessary, but not sufficient, for matrix targeting. A minimal NMTS can be generated by linking the rat GR DNA-binding domain to either its tau2 transactivation domain in its natural context, or a heterologous transactivation domain derived from the Herpes simplex virus VP16 protein. The transactivation and nuclear matrix-targeting activities of tau2 are separable, as transactivation mutants were identified that either inhibited or had no apparent effect on matrix targeting of tau2. A functional interaction between the NMTS of rat GR and the RNA-binding nuclear matrix protein hnRNP U was revealed in cotransfection experiments in which hnRNP U overexpression was found to interfere with the transactivation activity of GR derivatives that possess nuclear matrix-binding capacity. We have therefore ascribed a novel function to a steroid hormone transactivation domain that could be an important component of the mechanism used by steroid hormone receptors to regulate genes in their native configuration within the nucleus. PMID- 9731710 TI - BOD (Bcl-2-related ovarian death gene) is an ovarian BH3 domain-containing proapoptotic Bcl-2 protein capable of dimerization with diverse antiapoptotic Bcl 2 members. AB - Using the yeast two-hybrid protein-protein interaction system to search for genes capable of forming dimers with the antiapoptotic protein Mcl-1, we have isolated BOD (Bcl-2-related ovarian death agonist) from an ovarian fusion cDNA library. The three variants of BOD (long, medium, and short) have an open reading frame of 196, 110, and 93 amino acids, respectively; all of them contain a consensus Bcl-2 homology 3 (BH3) domain but lack other BH domains found in channel-forming Bcl-2 family proteins. In the yeast cell assay, BOD interacts with diverse antiapoptotic Bcl-2 proteins [Mcl-1, Bcl-2, Bcl-xL, Bcl-w, Bfl-1, and Epstein Barr virus (EBV) BHRF-1] but not with different proapoptotic Bcl-2 proteins (BAD, Bak, Bok, and Bax). After overexpression in mammalian Chinese hamster ovary (CHO) cells, BOD induces apoptosis that can be prevented by the baculoviral caspase inhibitor P35. The cell-killing activity of BOD is also antagonized in cells cotransfected with the antiapoptotic Bcl-w protein, which showed high affinity for BOD in the two-hybrid assay. Furthermore, mutagenesis studies showed that BOD mutants with alterations in the BH3 domain lose cell-killing ability, suggesting that the BH3 domain is important for the mediation of cell killing by BOD. BOD mRNA is ubiquitously expressed in ovary and multiple other tissues. The BOD gene is also conserved in diverse mammalian species. Identification of BOD expands the group of proapoptotic Bcl-2 proteins that only contains the BH3 domain and allows future elucidation of the intracellular mechanism for apoptosis regulation in ovary and other tissues. PMID- 9731711 TI - Functional antagonism of gonadal steroids at the 5-hydroxytryptamine type 3 receptor. AB - Steroid hormone action involves binding to cognate intracellular receptors that, in turn, bind to respective response elements and thus modulate gene expression. The present study shows that the gonadal steroids, 17beta-estradiol and progesterone, may also act as functional antagonists at the 5-hydroxytryptamine type 3 (5-HT3) receptor in whole-cell voltage-clamp recordings of HEK 293 cells stably expressing the 5-HT3 receptor. Functional antagonistic properties at this ligand-gated ion channel could also be shown for 17alpha-estradiol, 17alpha ethinyl-17beta-estradiol, mestranol, R 5020, testosterone, and allopregnanolone but not for pregnenolone sulfate and cholesterol. An antagonism at the 5-HT3 receptor could further be observed with the aromatic alcohol 4-dodecylphenol but not with phenol or ethanol. Thus, the modulation of 5-HT3 receptor function by steroids or alcohols is dependent on their respective molecule structure. The antagonistic action of steroids at the 5-HT3 receptor is not mediated via the serotonin binding site because the steroids did not alter the binding affinity of [3H]GR65630 to the 5-HT3 receptor, and kinetic experiments revealed a quite different response pattern to 17beta-estradiol when compared with the competitive antagonist metoclopramide. BSA-conjugated gonadal steroids labeled with fluorescein isothiocyanate bound to membranes of HEK 293 cells expressing the 5 HT3 receptor in contrast to native HEK 293 cells. However, there was no dose dependent displacement of the binding of gonadal steroids to membranes of cells expressing the 5-HT3 receptor in binding experiments or fluorescence studies. Thus, gonadal steroids probably interact allosterically with the 5-HT3 receptor at the receptor-membrane interface. The functional antagonism of gonadal steroids at the 5-HT3 receptor may play a role for the development and course of nausea during pregnancy and of psychiatric disorders. PMID- 9731712 TI - Insulin-like growth factor-I affects perinatal lethality and postnatal development in a gene dosage-dependent manner: manipulation using the Cre/loxP system in transgenic mice. AB - Insulin-like growth factor-I (IGF-I) is essential for cell growth, differentiation and postnatal development. A null mutation in igf-1 causes intrauterine growth retardation and perinatal lethality. The present study was designed to test the lower limit of igf-1 gene dosage that ensures survival and postnatal growth by using the Cre/loxP system. Mice with variable reductions in IGF-I levels were generated by crossing EIIa-cre transgenic mice and mice with loxP-flanked igf-1 locus (igf-1/flox). EIIa-cre mice express bacteriophage P1 Cre (causes recombination) recombinase under the adenovirus promoter EIIa, during early embryonic development before implantation, and cause genomic recombination of the igf-1/flox locus. Mice with the most extensive recombination die immediately after birth, while the survivors have significant growth retardation in proportion to the reduction in their igf-1 gene. Interestingly, this gene dosage effect on body weight was not very significant before weaning. However, when the young animals were weaned at 3 weeks, the igf-1 gene dosage was the only independent predictor of the weight gain between 3 and 6 weeks among the parameters tested. Although growth retarded, mice with Cre-induced partial igf-1 deficiency were fertile and gave birth to null mice. Thus Cre-induced genomic recombination using the EIIa promoter occurs during development and creates distinct phenotypes compared with the conventional null mutation. This variability allows for postnatal survival and will enable one to begin to explore the role of the endocrine vs. paracrine effects of IGF-I. PMID- 9731713 TI - In memoriam: Roy Orval Greep. PMID- 9731714 TI - Hypothesis: comparisons of inter- and intra-individual variations can substitute for twin studies in drug research. AB - Twin studies are useful devices to determine the heritability of persistent but variable characteristics that tend to differ among individuals. Drug responses are not persistent affairs; they are temporary characteristics. One therefore may ask whether twin studies are necessary to assess the genetic element in pharmacological responsiveness. To measure the genetic component contributing to their variability, it seems logical to investigate the response variation by repeated drug administration to given individuals, and to compare the variability of the responses within and between individuals. We attempt here to describe a theoretical background of this venture, and to show some results of the exercise. Potential sources of error or uncertainty are discussed. PMID- 9731715 TI - N-acetyltransferase NAT1 and NAT2 genotypes and lung cancer risk. AB - Acetyltransferases, encoded by the NAT1 and NAT2 genes, are involved in the activation/inactivation reactions of numerous xenobiotics, including tobacco derived aromatic amine carcinogens. Several allelic variants of NAT1 and NAT2, which cause variations in acetylation capacity, have been detected. The NAT2 slow acetylator phenotype/genotype has been inconsistently associated with lung cancer and, to date, the role of NAT1 polymorphism in lung cancer has not been reported. The effect of NAT1 and NAT2 genetic polymorphisms on individual lung cancer risk was evaluated among 150 lung cancer patients and 172 control individuals, all French Caucasian smokers. The NAT1 alleles (*3, *4, *10, *11, *14, and *15) and the NAT2 alleles (*4, *5, *6, *7) were differentiated by polymerase chain reaction-based restriction fragment length polymorphism methods using DNA extracted from peripheral white blood cells. Genotypes were classified according to current knowledge of the functional activity of the variant alleles. The NAT1*10 and NAT1*11 alleles were considered as rapid alleles, the NAT1*4 and the NAT1*3 as normal alleles and NAT1*14 and NAT1*15 as slow-acetylation alleles. Logistic regression analyses were performed taking into account the age, sex, smoking and occupational exposures. A significant association was observed between lung cancer and NAT1 genotypes (P(homogeneity) < 0.02) with a gene dose effect (P(trend) < 0.01); compared with homozygous rapid acetylators, the lung cancer risk was 4.0 (95% confidence interval 0.8-19.6) for heterozygous rapid acetylators, 6.4 (95% confidence interval 1.4-30.5) for homozygous normal acetylators and 11.7 (95% confidence interval 1.3-106.5) for heterozygous slow acetylators. None of the individuals were homozygous slow acetylators. Similar results were obtained whatever the adjustment considered. No significant association was found between NAT2 genotype and lung cancer. The NAT1 polymorphism may thus be an important modifier of individual susceptibility to smoking-induced lung cancer. PMID- 9731716 TI - Detection of mutations and polymorphism of N-acetyltransferase 1 gene in Indian, Malay and Chinese populations. AB - The xenobiotic metabolizing enzymes N-acetyltransferases (NATs) are important for the biotransformation and/or bioactivation of drugs and carcinogens. NATs are coded for in humans by two distinct genes, designated NAT1 and NAT2. NAT1, which was originally thought to be monomorphic, was recently reported to exhibit variation in human populations. Recent studies suggested that a genetic polymorphism of NAT1 may be associated with colorectal cancer risk. The distributions of NAT1 allele and genotype frequencies in unrelated individuals among Indian (n = 140), Malay (n = 122) and Chinese (n = 181) populations in Singapore were characterized by polymerase chain reaction-restriction fragment length polymorphism and allele-specific-polymerase chain reaction. The allelic frequencies of NAT1*3, NAT1*4, NAT1*10 and NAT1*11 among Indians were 0.3, 0.51, 0.17 and 0.02, respectively. The corresponding NAT1 allelic frequencies in Malays were 0.29, 0.30, 0.39 and 0.02, respectively, and were similar to those in Chinese in the region. The allelic frequencies of NAT1*3, NAT1*4, NAT1*10 and NAT1*11 among Chinese were 0.33, 0.35, 0.30 and 0.02, respectively. These findings are of importance for the determination of cancer risk in these populations. In addition, nucleotide changes at positions 350-351 (GG to CC) and 497-499 (GGG to CCC) of the NAT1 gene were not found in the alleles of the populations studied. PMID- 9731717 TI - NAD(P)H:quinone oxidoreductase: polymorphisms and allele frequencies in Caucasian, Chinese and Canadian Native Indian and Inuit populations. AB - NAD(P)H:quinone oxidoreductase (NQO1) catalyses the two-electron reduction of quinone compounds. NQO1 is involved in the reductive bioactivation of cytotoxic antitumour quinones such as mitomycin C, but also plays a protective role against the carcinogenicity and mutagenicity of quinones, their precursors and metabolites. Three alleles have been identified in the human population: the functional Arg139/Pro187 allele (which we have termed NQO1*1); the nonfunctional allele Arg139/Ser187 (NQO1*2) and the Trp139/Pro187 allele (NQO1*3), which is associated with a diminished activity. We applied polymerase chain reaction-based genotyping assays to characterize interethnic variability in the frequency of NQO1 alleles in Caucasian (n = 575), Canadian Native Indian (n = 110), Canadian Inuit (n = 83) and Chinese (n = 86) populations. The NQO1*2 allele was found at significantly higher frequencies in Chinese (0.49) and Native North American populations (Inuit 0.46; Canadian Native Indians 0.40) compared with Caucasians (0.16). The NQO1*3 allele was not observed in Inuit individuals, and occurred at a lower frequency than the NQO*2 allele in Caucasians (0.05), Chinese (0.04) and Canadian Native Indians (0.01). Our results predict that a greater proportion of Orientals and related ethnic groups lack, or have reduced, NQO activity relative to Caucasians. Affected individuals may not only exhibit resistance to quinone based cancer therapy because of a decreased production of cytotoxic drug metabolites, but may also be more susceptible to toxicities associated with toxicants. PMID- 9731718 TI - The relationship between CYP1A1 aryl hydrocarbon hydroxylase activity and lung cancer in a Japanese population. AB - Because aryl hydrocarbon hydroxylase (AHH) is considered to be responsible for the activation of benzo(a)pyrene and other polyaromatic hydrocarbons in cigarette smoke to carcinogens, it is important to examine CYP1A1 (AHH) activity in the determination of susceptibility to lung cancer. We investigated AHH activity in peripheral mitogen-treated lymphocytes in 108 lung cancer patients and 95 healthy control individuals. Non-induced AHH activity was detectable in all the samples. AHH inducibility (3-methylcholanthrene-induced/non-induced AHH activity) showed a very wide interindividual variation as well as non-induced AHH activity. No significant associations were found between adjusted AHH activity and histologic type of tumor among lung cancer patients. Adjusted AHH inducibility of genotype C [geometric mean and 95% confidence interval (CI); 15.56 and 11.69-20.71] in MspI polymorphism was significantly higher than those of the other two genotypes (P = 0.0001), while no significant difference was observed between genotypes A (4.76 and 3.82-5.93) and B (5.60 and 4.57-6.86). On the other hand, non-induced AHH activity of genotype Val/Val (0.121 and 0.082-0.178 pmol/min/10(6) cells) in isoleucine-valine (Ile-Val) polymorphism was significantly higher than those of genotypes Ile/Ile (0.042 and 0.034-0.052 pmol/min/10(6) cells) and Ile/Val (0.040 and 0.030-0.053 pmol/min/10(6) cells) (P < 0.0001). Even after controlling for age, cigarettes smoked per day and season of the year, high AHH inducibility (7.0 < versus 0 < < or = 3.0: OR and 95 %CI, 12.4 and 2.88-53.4) was an independent risk factor for lung cancer. The data indicate that high AHH inducibility may strongly associate with the susceptibility to lung carcinogenesis. PMID- 9731719 TI - Comparison of three CYP2D6 probe substrates and genotype in Ghanaians, Chinese and Caucasians. AB - The ability to metabolize CYP2D6 substrates sparteine, debrisoquine, and dextromethorphan was studied in healthy Caucasian (n = 20), Ghanaian (n = 21), and Chinese (n = 22) CYP2D6 extensive metabolizers. Genotype analysis for the CYP2D6*1, *3, *4, *5, *9, *10, and *17 alleles was performed. Interethnic differences in the disposition of the probe drugs were found among the extensive metabolizers; extensive metabolizer status was confirmed by phenotype and genotype analysis. The mean metabolic rate was lower for Caucasians than for Ghanaians for sparteine (P < 0.02) and for both Ghanaians and Chinese for debrisoquine (P < 0.02). Correlation comparisons resulted in lower pairwise correlation coefficients in Ghanaians compared with Chinese and Caucasians for every combination of probe substrates. In addition, in Chinese and Caucasians, metabolic rates for each pair of probe drugs were significantly correlated (P < 0.002), but in Ghanaians the dextromethorphan metabolic rates were not correlated to either sparteine or debrisoquine (P < 0.05). Even when only those with a CYP2D6*1/*1 genotype were included in the correlation calculations, the Ghanaians had very low correlation coefficients (r(s) - 0.02-0.2, n = 9); much lower than those found in Caucasian (r(s) 0.78-0.92, n = 14) or Chinese (r(s) 0.54-0.96, n = 7) individuals. Quinidine had significantly less affect on sparteine metabolic rates in Ghanaians than both Caucasians and Chinese (P < 0.02). In addition, five of the 21 Ghanaian individuals had dextromethorphan metabolic ratios which were unaffected by quinidine. These individuals also had differences in urinary recovery of dextromethorphan and its metabolites when compared to the other Ghanaian individuals. These results confirm the large ethnic differences in probe drug metabolism and quinidine sensitivity among these ethnic groups. They also suggest that the Ghanaians have an additional unidentified allele(s) with altered substrate specificity and quinidine sensitivity which is currently genotyped as CYP2D6*1. PMID- 9731720 TI - The RsaI polymorphism of CYP2E1 and susceptibility to alcoholic liver disease in Caucasians: effect on age of presentation and dependence on alcohol dehydrogenase genotype. AB - Twin studies in Caucasians suggest that susceptibility to alcoholic liver disease is, in part, genetically determined. Because most of the deleterious effects of alcohol are caused by its metabolism, attention has focused upon genes encoding ethanol metabolizing enzymes. Caucasians are polymorphic at only two of these gene loci--cytochrome P450 2E1 (CYP2E1) and alcohol dehydrogenase 3 (ADH3). We examined the frequency of the RsaI polymorphism of CYP2E1 and ADH3 genotype in 264 patients with alcoholic liver disease and 121 local control individuals. There was a non-significant excess of the rare c2 CYP2E1 allele in patients with advanced liver disease compared with control individuals/patients with steatosis only (0.029 versus 0.017/0.00). However, patients with the c2 allele presented at a younger age compared with those with the wild type c1 allele only (42.3 +/- 1.6 years versus 49.0 +/- 0.6 years; P = 0.001) with at least as advanced histology (93% cirrhotic versus 74%). Male patients had a higher frequency of the ADH3*2/*2 genotype (which encodes the less active gamma2 subunit) than control individuals [odds ratio (OR) 2.04 (1.11-3.76)], however, ADH3 genotype did not differ with histological stage or with age of presentation. Patients with advanced disease possessing the c2 allele had a significantly higher frequency of the ADH3*2/*2 genotype compared with c1 homozygotes [OR 3.71 (1.24-11.09)]. This study demonstrates that, although rare in Caucasians, possession of the mutant c2 allele of CYP2E1 increases the risk of alcoholic liver disease at a given level of cumulative alcohol consumption. This risk appears to be particularly manifest in individuals carrying the ADH3*2 allele, presumably reflecting increased metabolism of ethanol by CYP2E1. In the absence of the c2 allele, ADH3 genotype does not influence the risk of advanced alcoholic liver disease but, in males at least, may influence the risk of alcoholism. PMID- 9731721 TI - Genetic polymorphisms of CYP2D6, CYP1A1 and CYP2E1 in the South-Amerindian population of Chile. AB - Polymorphisms of cytochrome P450 genes show pronounced interethnic variation and have not been previously studied in the South-Amerindian population, which probably has an Asian origin. Therefore, a similar distribution of allelic and haplotype frequencies of cytochrome P450 genes to Asian populations might be expected in South-Amerindians. We analysed the allelic frequencies and haplotype distribution for CYP2D6, CYP1A1 and CYP2E1 genes in the South-Amerindian population of Chile (Mapuche, n = 84) by Southern blot or polymerase chain reaction-restriction fragment length polymorphism. Similar allelic frequencies and haplotype distribution for the CYP2E1 gene between Mapuches and Asian populations were observed. Frequencies of the two major functional CYP2D6*1 and CYP2D6*2 alleles and the CYP2D6*5 null allele were similar to most populations world-wide. The alleles CYP2D6*3 and *9, absent in Asians, were not found in Mapuches. The CYP2D6*4 allelic group, uncommon in Asian populations, had a low frequency in Mapuches (0.036). However, the CYP2D6*10 allele (Ch1, Ch2 and J), highly frequent in Asians (0.33-0.50), had a very low frequency (0.018) in our study population. In addition, the presence of the common Chinese 44 kb XbaI fragment of CYP2D6 (0.19-0.31 in Asians) was not detected in South-Amerindians. Interestingly, high frequencies for the rare m2 and Val alleles of the CYP1A1 gene were found in Mapuches (0.821 and 0.91, respectively), and the rare Val/m2 haplotype was significantly higher in Mapuches (0.748) than in Asians (0.24) (P < 0.01). The frequency of this haplotype in Mapuches is the highest frequency reported to date. The population studied was in Hardy-Weinberg equilibrium for these polymorphisms. The major differences between Mapuches and Asians were for CYP2D6*10 and CYP1A1 allelic frequencies, as well as the absence of the common Chinese 44 kb XbaI fragment of CYP2D6. These differences might be interpreted as a consequence of genetic drifts caused by a founder effect in the settlement of South-Amerindians, or genetic selection caused by dietary or environmental factors. PMID- 9731722 TI - No association between tacrine transaminitis and the glutathione transferase theta genotype in patients with Alzheimer's disease. PMID- 9731723 TI - The UGT1A1*28 allele is relatively rare in a Japanese population. PMID- 9731724 TI - Characterization of polymorphisms IVS25AS-6A to G, C1237T and T3409C in the human angiotensin converting enzyme gene. PMID- 9731725 TI - Biomarkers in benign breast disease: risk factors for breast cancer. PMID- 9731727 TI - Defects in mitotic checkpoint genes may speed cells toward malignancy. PMID- 9731726 TI - Increasing incidence of childhood primary malignant brain tumors--enigma or no brainer? PMID- 9731728 TI - Smoke this: genetics research begins to uncloud who gets hooked. PMID- 9731729 TI - Spirituality returns to the fold in medical practice. PMID- 9731730 TI - How randomized clinical trials came into their own. PMID- 9731731 TI - Journal letters financed by tobacco industry. PMID- 9731732 TI - Immunohistochemical detection of c-erbB-2 and p53 in benign breast disease and breast cancer risk. AB - BACKGROUND: We studied the associations between c-erbB-2 protein overexpression and p53 protein accumulation in benign breast tissue and the risk of subsequent breast cancer. METHODS: We conducted a case-control study nested within the cohort of 4888 women in the National Breast Screening Study (NBSS) who were diagnosed with benign breast disease during active follow-up. Case subjects were the women who subsequently developed breast cancer (ductal carcinoma in situ [DCIS] or invasive carcinoma). Control subjects were matched to each case subject on NBSS study arm, screening center, year of birth, and age at diagnosis of benign breast disease. Histologic sections of benign and cancerous breast tissues were analyzed immunohistochemically. Information on potential confounding factors was obtained by use of a self-administered lifestyle questionnaire. RESULTS: Accumulation of p53 protein was associated with an increased risk of progression to breast cancer (adjusted odds ratio [OR] = 2.55; 95% confidence interval [CI] = 1.01-6.40), whereas c-erbB-2 protein overexpression was not (adjusted OR = 0.65; 95% CI = 0.27-1.53). The findings for c-erbB-2 and p53 did not differ among strata defined by menopausal status, allocation within the NBSS, history of breast disease, and whether the benign breast disease was detected at a scheduled screen or between screens. The results were also similar after exclusion of case subjects whose diagnosis of breast cancer occurred within 1 year of their diagnosis of benign breast disease and after exclusion of subjects with DCIS. CONCLUSIONS: p53 protein accumulation, but not c-erbB-2 protein overexpression, appears to be associated with an increased risk of progression to breast cancer in women with benign breast disease. PMID- 9731733 TI - Trends in reported incidence of primary malignant brain tumors in children in the United States. AB - BACKGROUND: The reported incidence of primary malignant brain tumors among children in the United States increased by 35% during the period from 1973 through 1994. The purpose of our study was twofold: 1) to determine whether the reported incidence rates for this period are better represented by a linear increase over the entire period ("linear model") or, alternatively, by a step function, with a lower rate in the years preceding 1984-1985 and a constant higher rate afterward ("jump model"); and 2) to identify the specific brain regions and histologic subtypes that have increased in incidence. METHODS: Incidence data from the Surveillance, Epidemiology, and End Results Program of the National Cancer Institute for the period from 1973 through 1994 for primary malignant brain tumors in children were used to model the number of cases in a year as a random variable from a Poisson distribution by use of either a linear model or a jump model. RESULTS/CONCLUSIONS: The increase in reported incidence of childhood primary malignant brain tumors is best explained by the jump model, with a step increase in incidence occurring in the mid-1980s. The brain stem and the cerebrum are the primary sites for which an increase in tumor incidence has been reported. The increase in reported incidence of low-grade gliomas in the cerebrum and the brain stem (unaccompanied by an increase in mortality for these sites) supports the substantial contribution of low-grade gliomas to the overall increase in reported incidence for childhood brain tumors. IMPLICATIONS: The significantly better fit of the data to a jump model supports the hypothesis that the observed increase in incidence somehow resulted from changes in detection and/or reporting of childhood primary malignant brain tumors during the mid 1980s. PMID- 9731734 TI - Oral psoralen and ultraviolet-A light (PUVA) treatment of psoriasis and persistent risk of nonmelanoma skin cancer. PUVA Follow-up Study. AB - BACKGROUND/METHODS: The treatment of psoriasis with high-dose exposure to oral psoralen and ultraviolet-A light (i.e., PUVA) substantially increases the risk of cutaneous squamous cell cancer, but not of basal cell cancer, within a decade of beginning treatment. To assess the persistence of cancer risk among individuals treated with PUVA, including those who discontinued therapy long ago and those without substantial exposure to other carcinogens, we prospectively studied a cohort of 1380 patients with psoriasis who were first treated during the period from January 1, 1975, through October 1, 1976, and evaluated risk factors associated with the development of cutaneous squamous cell cancers and basal cell cancers after 1985. RESULTS: From 1975 through 1996, 237 patients developed 1422 cutaneous squamous cell cancers. From 1986 through 1996, 135 (12.5%) of 1081 patients without a prior squamous cell cancer developed 593 such tumors. From 1975 through 1997, 247 patients developed 1042 basal cell cancers; these patients included 151 individuals with a first basal cell cancer after 1985. Among those without a squamous cell or a basal cell cancer in the first decade of the prospective study, a strong dose-related increase in the risk of squamous cell cancer was observed in the subsequent decade (adjusted relative risk [> or =337 treatments versus <100 treatments] = 8.6; 95% confidence interval = 4.9-15.2). Risk of basal cell cancer was substantially increased only in those patients exposed to very high levels of PUVA (> or =337 treatments). CONCLUSIONS: High dose exposure to PUVA is associated with a persistent, dose-related increase in the risk of squamous cell cancer, even among patients lacking substantial exposure to other carcinogens and among patients without substantial recent exposure to PUVA. Exposure to PUVA has far less effect on the risk of basal cell cancer. The use of PUVA for psoriasis should be weighed against the increased cancer risk. PMID- 9731735 TI - Distinct altered patterns of p27KIP1 gene expression in benign prostatic hyperplasia and prostatic carcinoma. AB - BACKGROUND: The p27KIP1 gene, whose protein product is a negative regulator of the cell cycle, is a potential tumor suppressor gene; however, no tumor-specific mutations of this gene have been found in humans. This study was undertaken to identify and to assess potential alterations of p27KIP1 gene expression in patients with benign prostatic hyperplasia (BPH) and patients with prostate cancer. METHODS: We analyzed 130 prostate carcinomas from primary and metastatic sites, as well as prostate samples from normal subjects and from patients with BPH. Immunohistochemistry and in situ hybridization were used to determine the levels of expression and the microanatomical localization of p27 protein and messenger RNA (mRNA), respectively. Immunoblotting and immunodepletion assays were performed on a subset of the prostate tumors. Associations between alterations in p27KIP1 expression and clinicopathologic variables were evaluated with a nonparametric test. The Kaplan-Meier method and the logrank test were used to compare disease-relapse-free survival. Prostate tissues of p27Kip1 null (i.e., knock-out) and wild-type mice were also evaluated. RESULTS: Normal human prostate tissue exhibited abundant amounts of p27 protein and high levels of p27KIP1 mRNA in both epithelial cells and stromal cells. However, p27 protein and p27KIP1 mRNA were almost undetectable in epithelial cells and stromal cells of BPH lesions. Furthermore, p27Kip1 null mice developed enlarged (hyperplastic) prostate glands. In contrast to BPH, prostate carcinomas were found to contain abundant p27KIP1 mRNA but either high or low to undetectable levels of p27 protein. Primary prostate carcinomas expressing lower levels of p27 protein appeared to be biologically more aggressive (two-sided P = .019 [Cox regression analysis]). CONCLUSIONS/IMPLICATIONS: On the basis of these results, we infer that loss of p27Kip1 expression in the human prostate may be causally linked to BPH and that BPH is not a precursor to prostate cancer. PMID- 9731737 TI - Cancer death rates associated with human immunodeficiency virus infection in the United States. PMID- 9731736 TI - Plasma sex steroid hormone levels and risk of breast cancer in postmenopausal women. AB - BACKGROUND: A positive relationship has generally been observed between plasma estrogen levels and breast cancer risk in postmenopausal women, but most of these studies have been small and few have evaluated specific estrogen fractions (such as percent bioavailable estradiol). In addition, few studies have evaluated plasma androgen levels in relation to breast cancer risk, and their results have been inconsistent. We prospectively evaluated relationships between sex steroid hormone levels in plasma and risk of breast cancer in postmenopausal women by use of a case-control study nested within the Nurses' Health Study. METHODS: Blood samples were collected during the period from 1989 through 1990. Among postmenopausal women not using hormone replacement therapy at blood collection (n = 11,169 women), 156 women were diagnosed with breast cancer after blood collection but before June 1, 1994. Two control subjects were selected per case subject and matched with respect to age, menopausal status, month and time of day of blood collection, and fasting status at the time of blood collection. RESULTS: From comparisons of highest and lowest (reference) quartiles, we observed statistically significant positive associations with risk of breast cancer for circulating levels of estradiol (multivariate relative risk [RR] = 1.91; 95% confidence interval [CI] = 1.06-3.46), estrone (multivariate RR = 1.96; 95% CI = 1.05-3.65), estrone sulfate (multivariate RR = 2.25; 95% CI = 1.23-4.12), and dehydroepiandrosterone sulfate (multivariate RR = 2.15; 95% CI = 1.11-4.17). We found no substantial associations with percent free or percent bioavailable estradiol, androstenedione, testosterone, or dehydroepiandrosterone. The positive relationships were substantially stronger among women with no previous hormone replacement therapy. CONCLUSION: Our data, in conjunction with past epidemiologic and animal studies, provide strong evidence for a causal relationship between postmenopausal estrogen levels and the risk of breast cancer. PMID- 9731738 TI - Re: Urokinase and the urokinase receptor: association with in vitro invasiveness of human bladder cancer cell lines. PMID- 9731739 TI - Dietary fat and breast cancer. PMID- 9731740 TI - Increased P-glycoprotein messenger RNA stability in rat liver tumors in vivo. AB - P-glycoproteins (Pgp) are comprised of a small family of plasma membrane proteins whose abundance in cultured cells is often associated with the multidrug resistance phenotype. Overexpression of Pgp has been observed in many types of human cancers, but the molecular basis for this overexpression has not been established. We have used primary monolayer cultures of adult rat hepatocytes and a stepwise model of rat liver carcinogenesis to study the regulation of Pgp gene expression. We observed a marked overexpression of Pgp, specifically the class II Pgp, in both systems. In addition, we observed that a number of unrelated genes including alpha-tubulin, beta-actin, gamma-actin, cytokeratin 8, cytokeratin 18, and c-myc are overexpressed in cultured hepatocytes, and they are also overexpressed during liver carcinogenesis and in transplantable tumors. Nuclear run-on assays showed no increase in the transcriptional activity of Pgp genes in transplantable liver tumors compared to normal liver. Studies of in vivo mRNA stability, however, revealed that all three Pgp mRNAs were relatively stable in transplantable liver tumors (t(1/2) > 12 h), in contrast to what was found in normal liver (t(1/2) < 2 h). In addition, mRNA for several other genes, including alpha-tubulin, c-myc, and cyclin D1, all appear to be stabilized in the tumors. These findings suggest that the overexpression of Pgp genes in rat liver tumors may be the result of a mechanism involving stabilization of a diverse group of mRNAs. PMID- 9731741 TI - Expression of insulin-like growth factor binding protein-5 by ovine granulosa cells is regulated by cell density and programmed cell death in vitro. AB - In vivo, in the sheep ovary, the expression of insulin-like growth factor binding protein (IGFBP)-2 and particularly IGFBP-5 has been shown to increase dramatically in apoptotic granulosa cells from atretic follicles. The aim of this work was to study the relationship between apoptosis induced by serum starvation in vitro and expression of IGFBP-2 and -5 by ovine granulosa cells. For this purpose, granulosa cells from follicles 1-3 mm in diameter were cultured in the presence of serum for 2 days, then cultured in the presence or absence of serum for 24, 48, or 72 hr. At the end of the culture, cells were counted, cell viability was assessed by studying DNA fragmentation, and IGFBPs expression was studied by quantitative autoradiography, Western-ligand blotting, immunoblotting, and quantitative in situ hybridization. In vitro, IGFBP-2 and particularly IGFBP 5 were the main IGFBPs secreted by ovine granulosa cells. Serum starvation provoked (i) apoptosis of granulosa cells within 48 hr, (ii) a marked decrease in cell density, and (iii) a marked increase in the amount of IGFBP-5 associated with cell membranes and with the walls of culture wells, but no change in culture medium. The increase in the amount of cell- and wall-associated IGFBP-5 after serum starvation was essentially due to the consecutive decrease in cell density rather than to an increase in cell apoptosis. Indeed, irrespective of the presence or absence of serum, the amount of IGFBP-5 associated to cell membranes was inversely correlated to cell density. In contrast, the amount of IGFBP-5 present in culture medium was positively correlated to cell density. Furthermore, expression of IGFBP-5 mRNA was shown to increase with both cell density and cell death. Indeed, the expression of IGFBP-5 mRNA dramatically increased with cell density, irrespective of the presence or absence of serum, but at a similar cell density, expression was higher in serum-free than in serum conditions. Overall, these results indicate that, in vitro, the localization of IGFBP-5 on ovine granulosa cell membranes and in culture medium, respectively, was mainly dependent on cell density, whereas expression of IGFBP-5 mRNA was related to both cell density and cell death. These data suggest that IGFBP-5 is involved in both growth arrest and apoptosis of granulosa cells in the sheep. PMID- 9731742 TI - Isolation and characterization of osteoclast precursors that differentiate into osteoclasts on calvarial cells within a short period of time. AB - Osteoclasts are formed in cocultures of mouse calvarial cells and hematopoietic cells in the presence of osteotropic factors such as 1alpha,25-dihydroxyvitamin D3 [1alpha,25(OH)2D3], parathyroid hormone (PTH) and prostaglandin E2 (PGE2). We isolated osteoclast precursors (OCPs) from the coculture and examined their characteristics. After coculture for 7 days of mouse calvarial cells and bone marrow cells in the absence of osteotropic factors, hematopoietic cells were recovered and applied to a Sephadex G-10 column. Cells which passed through the column were collected as OCPs. When OCPs were cultured on calvarial cell layers in the presence of 1alpha,25(OH)2D3, tartrate-resistant acid phosphatase (TRAP) positive cells first appeared within 24 h, and their number increased thereafter. OCPs also differentiated into TRAP-positive cells within 48 h on the calvarial cell layer which had been pretreated with either 1alpha,25(OH)2D3, PTH, or PGE2. Autoradiography using [125I]-labeled calcitonin showed that TRAP-positive cells formed on the calvarial cell layer expressed calcitonin receptors. Direct contact between OCPs and calvarial cells was required for the differentiation of OCPs into TRAP-positive cells. Flow cytometric analysis revealed that OCPs were positive for Mac-1, Mac-2, and Gr-1 but negative for F4/80, B220 and CD3e. Calvarial cells obtained from macrophage-colony stimulating factor (M-CSF) deficient osteopetrotic (op/op) mice did not support OCP formation. A cell preparation disaggregated from long bones of newborn mice contained OCPs that differentiated into TRAP-positive cells on calvarial cells within 48 h, but cell preparations of freshly isolated bone marrow cells and alveolar macrophages did not. These results suggest that OCPs are specific cells which are formed only in the bone microenvironment and that OCPs recognize a signal(s) expressed by stromal cells in response to osteotropic factors and differentiate into osteoclasts. PMID- 9731743 TI - Differential expression and biological activity of retinoic acid-induced TGFbeta isoforms in embryonic palate mesenchymal cells. AB - The effect of retinoic acid (RA) on TGF-beta mRNA expression and protein production in murine embryonic palate mesenchymal (MEPM) cells was examined by Northern blotting and TGF-beta bioassay in association with TGF-beta isoform specific neutralizing antibodies. Heat or acid activation was used to distinguish between latent and active TGF-beta protein released into the culture medium. RA had little or no effect on TGF-beta1 mRNA expression and protein production. In contrast, RA increased TGF-beta2 and beta3 protein released into the culture medium, the protein being mostly in an inactive or latent form. The amount of active TGF-beta released was increased relative to the total increase in TGF-beta released, suggesting that RA treatment stimulated activation of latent TGF-beta. RA also increased TGF-beta2 mRNA expression; we have previously shown that RA upregulates TGF-beta3 mRNA in these cells. RA and TGF-beta individually inhibited 3H-thymidine incorporation into MEPM cell DNA, while, when administered simultaneously, they inhibited proliferative activity to a greater extent. Heat- or acid-activated conditioned medium (CM) from MEPM cells treated with RA was able to inhibit 3H-thymidine incorporation into MEPM cell DNA to an extent greater than seen with RA treatment alone. Coincubation of heat-activated CM from RA-treated MEPM cells with pan-specific or TGF-beta2 or beta3-specific neutralizing antibodies partially relieved the inhibitory effect on 3H-thymidine incorporation, suggesting that this proliferative response was due to RA-induced TGF-beta. Simultaneous treatment with RA and TGF-beta also stimulated gycosaminoglycan (GAG) synthesis to an extent greater than that seen with TGF beta treatment alone, this despite the ability of RA to inhibit GAG synthesis. These data demonstrate a role for RA and RA-induced TGF-beta in the regulation of palate cell proliferation and GAG synthesis and suggest a role for TGF-beta in retinoid-induced cleft palate. PMID- 9731744 TI - Modulation of insulin-like growth factor actions in L6A1 myoblasts by insulin like growth factor binding protein (IGFBP)-4 and IGFBP-5: a dual role for IGFBP 5. AB - We have previously shown that the insulin-like growth factors (IGFs) stimulate both proliferation and differentiation of skeletal muscle cells in culture, and that these actions in L6A1 muscle cells may be modulated by three secreted IGF binding proteins (IGFBPs), IGFBP-4, -5, and -6. Since we found that the temporal expression pattern of IGFBP-4 and IGFBP-5 differed dramatically during the transition from proliferating myoblasts to differentiated myotubes, we undertook the current study to examine the effects of purified IGFBP-4 and IGFBP-5 on IGF stimulated actions in L6A1 muscle cells. As has been shown for other cell types, we found that IGFBP-4 had only inhibitory actions, inhibiting IGF-I and IGF-II stimulated proliferation and differentiation. In contrast, IGFBP-5 exhibited both inhibitory and stimulatory actions. When added in the presence of 30 ng/ml IGF-I, IGFBP-5 (250 ng/ml) inhibited all markers of the early proliferative response: the tyrosine phosphorylation of the cytoplasmic signaling molecules IRS-1 and Shc, the activation of the MAP kinases, ERK1 and 2, the elevation of c-fos mRNA, the early inhibition of the elevation in myogenin mRNA, and the increase in cell number. In contrast, IGFBP-5 stimulated all aspects of the myogenic response to IGF-I: the later rise in myogenin mRNA, the elevation of creatine kinase activity, and the fusion of myoblasts into myotubes. This dual response to IGFBP 5 was greatest when it was added at a molar ratio of IGFBP-5 to IGF-I of 2:1. In contrast, when IGFBP-5 was added in the presence of IGF-II, it inhibited both proliferation and differentiation. Neither IGFBP had any effect when added in the presence of R3 IGF-I, an analog with substantially reduced affinity for IGFBPs. Our results suggest that the role of IGFBP-4 is mainly to sequester excess IGFs, and thus inhibit all actions. IGFBP-5, however, is capable of eliciting a dual response, possibly due to its unique ability to associate with the cell membrane. PMID- 9731745 TI - Autocrine FGF signaling is required for vascular smooth muscle cell survival in vitro. AB - Expression of both basic fibroblast growth factor (bFGF) and FGF receptors (FGFR) by vascular smooth muscle cells suggests that autocrine FGF signaling mechanisms may have important functions. Inhibition of smooth muscle cell bFGF expression provokes apoptosis, suggesting that endogenous bFGF generates an anti-apoptotic signal. The purpose of this study was to determine whether the survival function of endogenous bFGF requires signaling through FGFR. A recombinant adenovirus encoding a truncated murine FGFR-1 lacking the kinase domain (DN-FGFR) efficiently expressed the transgene in cultured rat aortic smooth muscle cells. The truncated receptor acted in a dominant negative fashion to effectively prevent receptor-mediated signaling, assessed by phosphorylation of p42/p44 MAP kinase. Expression of DN-FGFR provoked apoptosis of SMC in a dose-dependent fashion that was insensitive to recombinant bFGF but could be rescued by platelet derived growth factor or epidermal growth factor. Heterologous growth factor rescue was inhibited by PD98059, an inhibitor of MEK (MAP kinase-kinase). These data demonstrate that inhibition of FGF receptor activation results in apoptosis and suggest that an intact autocrine FGF signaling loop is required for vascular smooth muscle cell survival in vitro. These findings also implicate the Ras/Raf/MEK/MAP kinase cascade in generating or sustaining the survival signal. The functional significance of an autocrine FGF signaling loop in non-transformed cells has important implications for cardiovascular development, remodeling and disease. PMID- 9731746 TI - Withdrawal of 2-mercaptoethanol induces apoptosis in a B-cell line via Fas upregulation. AB - Mouse lymphoid cell cultures are dependent on reducing agents in their culture medium to allow proliferation and survival of the cells. In the case of the mouse CD5+-pre-B cell line SPGM-1, withdrawal of 2-mercaptoethanol (2-ME) resulted in rapid inhibition of proliferation and subsequent cell death by apoptosis. The pathways leading to cell death by withdrawal of 2-ME or by incubation with ionomycin, a known inducer of apoptosis, were compared. Both kinds of stimulation resulted in apoptosis of the whole population, but cell death occurred with different kinetics. Only apoptosis induced by ionomycin was inhibited by coincubation with the phorbol ester PMA, while apoptosis induced by withdrawal of 2-ME was not. Overexpression of the human bcl-2 proto-oncogene in these cells delayed the death process induced by either method. SPGM-1xbcl-2 cells accumulated in the G0/G1 and G2/M cell cycle phases after removal of 2-ME from the medium, whereas treatment with ionomycin resulted in an arrest only in the G0/G1 transition. Interestingly, both stimuli induced the expression of the Fas receptor, but with different kinetics, while the Fas ligand (FasL) was expressed constitutively in SPGM-1 cells. These data demonstrate that withdrawal of 2-ME and incubation with ionomycin both induce rapid cell death by apoptosis, possibly mediated by an autocrine Fas/FasL loop. Although the initial pathways activated by the two forms of treatment must be different, they converge on a common level controlled by the anti-apoptotic gene product Bcl-2. PMID- 9731747 TI - Thrombin receptor expression and responsiveness of human monocytic cells to thrombin is linked to interferon-induced cellular differentiation. AB - Human thrombin has been shown to stimulate monocyte chemotaxis, phagocytosis, and interleukin (IL8) production, but the mechanisms responsible for stimulation are not well defined. In some cells, thrombin stimulation of proliferation appears to require both cleavage of the proteolytically activated receptor for thrombin (PAR1) and activation of a nonproteolytically activated thrombin receptor (N PAR), while in others activation of either receptor alone may be sufficient for stimulation. We, therefore, have initiated studies to address thrombin receptor expression and cell responsiveness to thrombin in interferon gamma (IFNgamma) differentiated and nondifferentiated U937 monocytic cells. Northern blot analysis shows that PAR1 expression is upregulated upon differentiation. Experiments with biotinylated and 125I-thrombin show that specific thrombin binding is dramatically increased by differentiation although it is not clear if this binding is to PAR1 or to a separate binding component such as N-PAR which is present on fibroblasts and other cells. Addition of thrombin at concentrations of 1-10 microg/ml (30-300 nM, concentrations where specific thrombin binding is observed) stimulates proliferation of IFNgamma-differentiated U937 cells but not of undifferentiated U937 cells. Thrombin also stimulates interleukin-6 (IL6) production in IFNgamma-differentiated U937 cells. Moreover, thrombin induces high levels of IL6, interleukin-1beta (IL1beta), and tumor necrosis factor-alpha (TNF alpha) production by peripheral blood mononuclear cells (PBMC) and monocytes. These results show that differentiated U937 cells and mature PBMC are responsive to thrombin whereas nondifferentiated U937 are not. Further, this responsiveness appears to correlate with expression of PAR1 and to a dramatic increase in specific thrombin binding. That thrombin stimulates cytokine production and proliferation in populations of differentiated monocytes suggests that thrombin may be an important regulator of inflammation and wound healing. PMID- 9731748 TI - Isozymes of cyclic AMP-dependent protein kinases (PKA) in human lymphoid cell lines: levels of endogenous cAMP influence levels of PKA subunits and growth in lymphoid cell lines. AB - Activation of the cAMP signaling pathway in lymphoid cells is known to inhibit cell proliferation of T and B cells as well as cytotoxicity of natural killer (NK) cells. In order to find suitable model systems to study cAMP-mediated processes, we have examined the expression of cAMP-dependent protein kinase (PKA), endogenous levels of cAMP, and cell proliferation in eight cell lines of B lineage origin, four cell lines of T lineage origin, and normal human B and T cells. We demonstrated that the expression of mRNA and protein for one of the regulatory (R) subunits of PKA (RIalpha) was present in all the cells investigated, in contrast to the other R subunits (RIbeta, RIIalpha, and RIIbeta). Furthermore, three T cell lines and one B cell line expressed only RIalpha and C, implying these cells to contain solely PKA type I. Moreover, for the RI subunit, we observed an apparent reciprocal relationship between levels of mRNA and protein. Generally, RIalpha protein was low in cell lines where mRNA was elevated and vice versa. This was not the case for the RII subunits, where high levels of mRNA were associated with elevated levels of protein. Interestingly, we demonstrated an inverse correlation between levels of endogenous cAMP and cell growth as determined by [3H]-thymidine incorporation and cell-doubling rate (P < 0.05). Taken together, our results demonstrate great differences in PKA isozyme composition, which should be taken into consideration when using lymphoid cell lines as model system for cAMP/PKA effects in normal lymphocytes. PMID- 9731749 TI - Regulation of glucose transport by angiotensin II and glucose in cultured vascular smooth muscle cells. AB - Glucose transport in response to angiotensin II (AII) was assessed in cultured vascular smooth muscle (VSM) cells by measuring the uptake of [3H]-2 deoxyglucose, a radiolabeled non-metabolizable glucose analog. Significant stimulation occurred by 2 hr of exposure with the maximum effect being observed between 6 and 8 hr. All effects were concentration dependent with a threshold response being detected at 0.1 nM. AII-stimulated transport was blocked by saralasin, an AII receptor antagonist, indicating that AII binding to a specific receptor is required for AII to elicit the transport response. AII-stimulated transport was also blocked when cells were incubated with cycloheximide for 6 hr, suggesting that protein synthesis is required for the long-term effects of AII on glucose transport. A specific protein synthesized in response to AII stimulation was the GLUT 1 glucose transporter as assessed by western blot analysis. Inhibition of protein kinase C (PKC) by bisindolylmaleimide and staurosporine did not affect VSM responsiveness to AII, suggesting that AII is capable of stimulating glucose transport through a PKC-independent mechanism; however, VSM responsiveness to AII did appear to be dependent upon the presence of extracellular calcium. The importance of calmodulin in mediating the response of VSM cells to AII was indicated by the inhibition of AII-stimulated glucose transport when VSM cells were incubated in the presence of the calmodulin inhibitors, calmidazolium and W7. Finally, glucose uptake increased with decreasing levels of glucose in the incubation medium. This was accompanied by a corresponding decrease in the relative effectiveness of AII in stimulating glucose uptake. PMID- 9731750 TI - Potentiating effects of pertussis toxin on leukotriene C4 induced formation of inositol phosphate and prostacyclin in human umbilical vein endothelial cells. AB - Leukotriene C4 is an arachidonic acid metabolite and an important mediator of inflammation and anaphylaxis that is known to induce production of prostacyclin in endothelial cells. The goal of this study was to examine the signal transduction mechanisms activated by leukotriene C4 stimulation. Formation of inositol phosphates was measured to determine the activation of phospholipase C and pertussis toxin was used to explore the role of G-proteins. Additionally, we evaluated the role of protein kinase C in these events, especially whether there was an interaction between pertussis toxin mediated effects and the activity of protein kinase C. Leukotriene C4 induced a dose- and time-dependent formation of inositol phosphates and prostacyclin. The response to leukotriene C4 was greater than the response to leukotriene D4 even after treatment with L-serine borate complex, suggesting the presence of a specific leukotriene C4 receptor. Exposure to pertussis toxin potentiated, time-dependently, the leukotriene C4 induced formation of inositol phosphates and prostacyclin through a mechanism which was altered by manipulation of protein kinase C activity. The exact mechanism is not clear but our results are consistent with a postulated dual mechanism of phospholipase C control, in which leukotriene C4 induced stimulation is attenuated by a pertussis toxin sensitive G-protein. PMID- 9731751 TI - Activation of the sodium/hydrogen exchanger via the fibronectin-integrin pathway results in hematopoietic stimulation. AB - The proliferative response of hematopoietic cells is regulated by many factors, including the presence and type of growth factors, the cellular microenvironment, and the physiochemical conditions prevailing in the tissue milieu. A process fundamental to all cells is the regulation of the intracellular acid-base conditions. One of the mechanisms by which intracellular pH (pHi) is regulated is through the sodium/hydrogen exchanger, a ubiquitous membrane protein which exploits the intra- and extracellular sodium ion gradient to drive hydrogen ions out of the cell. However, activation of the exchanger via mitogenic and nonmitogenic signals leads to an increase in pHi which, in turn, may directly or indirectly result in a proliferative response. It has been shown that interaction of fibronectin with its integrin receptor subunits alpha4 and alpha5 can result in activation of the Na+/H+ exchanger. In this report, we demonstrate that when mouse bone marrow cells are physically brought together in a preculture system we designate as high cell density culture (HCDC), in a small volume and at the same cellularity as that in the marrow, hematopoietic stem and progenitor cell populations are stimulated with no additional stimulation in the presence of growth factors. Neutralizing antibodies to the growth factors added to HCDC had little, if any, effect on the degree of stimulation. However, when antibodies to fibronectin or the alpha4 integrin subunit were added to HCDC, inhibition was observed, indicating that the observed hematopoietic stimulation occurred via the fibronectin-integrin pathway. Addition of 5 microM 5-(N,N-hexamethylene) amiloride (5-HMA), a specific inhibitor of the Na+/H+ exchanger, also resulted in inhibition of in vitro hematopoiesis. Since the exchanger was implicated, we then measured the pHi of normal and HCDC-treated bone marrow cells in the absence and presence of 5-HMA by flow cytometry using the fluorescent pH-sensitive indicator, carboxy SNARF-1 AM. It was found that cells subjected to HCDC exhibited a higher pH than normal fresh cells. In each case, the pH was lowered in the presence of 5 HMA. Furthermore, addition of antibodies to fibronectin or the alpha4 integrin subunit to HCDC also reduced the pH, to a similar level to that found for 5-HMA. Our results demonstrate, for the first time, that a hematopoietic stem and progenitor cell proliferative response can be initiated by activation of the Na+/H+ exchanger, leading to an increase in pHi, via cell-cell interaction through the fibronectin-integrin pathway. This pathway could, therefore, be significant not only in normal hematopoietic regulation, but also under pathophysiological conditions. PMID- 9731752 TI - Focal delivery of fibroblast growth factor-1 by transfected cells induces spiral ganglion neurite targeting in vitro. AB - Sensory cells in the cochlea of the rat transiently express acidic fibroblast growth factor (FGF-1) during the developmental period of terminal innervation in the sensory epithelium. To explore the potential role of FGF-1 in terminal innervation events, the response of cochlear ganglion neurons to FGF-1 was evaluated in culture. Explants from the spiral ganglion of postnatal day 5 rats were cultured in the presence of exogenous FGF-1, with or without heparin. FGF-1 in the culture medium produced a dose-dependent increase in the number and length of neurites produced by spiral ganglion neurons, a response that was enhanced by heparin. To assess the effects of FGF-1 produced by a focal, cellular source, additional explants were cocultured with 3T3 cell transfectants that secrete FGF 1. Neurites that came into contact with FGF-1 secreting cells branched, formed bouton-like terminal swellings on the surface of the transfectants, and stopped extending. The results suggest that FGF-1 may stimulate neurite extension into the sensory epithelium of the cochlea and that focal production of FGF-1 may contribute to the formation of contacts on sensory cells by developing neurites. PMID- 9731753 TI - Possible involvement of p21/waf1 in the growth inhibition of HepG2 cells induced by hepatocyte growth factor. AB - Hepatocyte growth factor (HGF) is a potent mitogen for a variety of cell types, but it is also known as an antimitogenic factor for several types of tumor cell lines. The biological processes by which HGF inhibits tumor cell growth remain poorly understood. Here we report a comparative study of HGF-mediated signal transduction events between two opposite responding types of human hepatoblastoma cell lines, HuH6 and HepG2. Following serum starvation, both cell lines were cultured in hepatocyte growth medium (HGM), a chemically defined medium, in the presence or absence of HGF. Under these culture conditions, cell growth in HuH6 was promoted by HGF, while it was inhibited in HepG2. Phosphorylation of p42/mitogen-activated protein (MAP) kinase was observed within 10 min after HGF stimulation in both cell lines. The level of phosphorylated MAP kinase in HuH6 declined to basal levels after 2 hr. However, in HepG2 the phosphorylated form was detectable at 6 hr. p21/waf1 was induced in both cell lines where levels peaked 4-6 hr after HGF stimulation. In HuH6, a marked decrease of p21/waf1 was observed at 8-12 hr, while a high level of p21/waf1 was sustained for at least 24 hr in HepG2. HGF treatment depressed cdk2 activity in a time-dependent manner in HepG2 while the activity increased in HuH6. When serum-starved HepG2 was growth stimulated with serum in the presence or absence of HGF, the cells treated with HGF underwent growth inhibition correlating with a sustained induction of p21/waf1 and a decrease of cdk2 activity. Immunoprecipitation analysis revealed accumulation of cdk2-associated p21/waf1 in the HGF-treated HepG2. Together, the results suggest that sustained induction of p21/waf1 mediates growth inhibition in HepG2 in the presence of HGF. PMID- 9731754 TI - Tumor cell apoptosis, lymphocyte recruitment and tumor vascular changes are induced by low temperature, long duration (fever-like) whole body hyperthermia. AB - A single treatment of low-temperature, long-duration, whole-body hyperthermia of either severe combined immunodeficient (SCID) mice bearing human breast tumor xenografts or Balb/c mice bearing syngeneic tumors for 6-8 hr can cause a temporary reduction of tumor volume and/or a growth delay. In both animal model systems, this inhibition is correlated with the appearance of large numbers of apoptotic tumor cells. Because this type of mild heat exposure, comparable to a common fever, is not itself directly cytotoxic, other explanations for the observed tumor cell death were considered. Our data support the hypothesis that this hyperthermia protocol stimulates some component(s) of the immune response, which results in increased antitumor activity. In support of this hypothesis, increased numbers of lymphocyte-like cells, macrophages, and granulocytes are observed in the tumor vasculature and in the tumor stroma immediately following this mild hyperthermia exposure. In Balb/c mice, an infiltrate persists in the tumor for at least 2 weeks. Using the SCID mouse/human tumor system, we found that both host natural killer (NK) cells and injected human NK cells were increased at the site of tumor following hyperthermia treatment. Experiments using anti-asialo-GM1 antibodies indicate that the tumor cell apoptosis seen in the SCID mouse appears to be due largely to the activity of NK cells, although additional roles for other immunoeffector cells and cytokines appear likely in the immunologically complete Balb/c model. Another interrelated hypothesis is that immunoeffector cells may have greater access to the interior of the tumor because we have observed that this treatment causes an obvious expansion in the diameter of blood vessels within the tumor and an increase in nucleated blood cells within the vessels, which persists as long as 2 weeks after treatment. Further study of the mechanisms by which mild hyperthermia exerts antitumor activity could result in this treatment protocol being used as an effective, nontoxic adjuvant to immunotherapy and/or other cancer therapies. PMID- 9731755 TI - Modulation of antioxidant enzymes, reactive oxygen species, and glutathione levels in manganese superoxide dismutase-overexpressing NIH/3T3 fibroblasts during the cell cycle. AB - NIH/3T3 mouse embryo fibroblasts were transfected with the cDNA for manganese superoxide dismutase (MnSOD). Previous studies showed characteristic unique AE profiles in nonsynchronous populations of parental, control plasmid-transfected, and MnSOD-overexpressing NIH/3T3 cell lines. However, the present study showed that during S and M phases of the cell cycle, antioxidant enzyme (AE) levels were altered in MnSOD-overexpressing cell lines towards levels in S and M phases of parental and control plasmid-transfected cells. Because of the demonstration that MnSOD overexpression inhibits cell growth in both nonmalignant and malignant cells, the present study was designed to measure AEs, reactive oxygen species (ROS), and glutathione levels in various phases of the cell cycle in both parental NIH/3T3 cells and NIH/3T3 cells overexpressing MnSOD, to try to determine whether AEs, ROS, and glutathione levels could have a possible regulatory role in cell cycle progression. In all cell lines studied, ROS levels were lower in M than S phase of the cell cycle. Total glutathione and glutathione disulfide levels were greatly increased during the M phase of the cell cycle compared with quiescence and S phase in all cell lines studied. This suggests that oxidative stress exists in M phase of the cell cycle with total glutathione levels increased to decrease oxidative stress. Analysis of MnSOD-overexpressing cell clones showed a correlation of decreased cell growth with an increase in ROS in S phase of the cell cycle and a decrease in glutathione in mitosis. The data strongly suggest that specific levels of cell redox state are necessary for cells to successfully progress through the various phases of the cell cycle. PMID- 9731756 TI - Effects of polyamines and calcium and sodium ions on smooth muscle cytoskeleton associated phosphatidylinositol (4)-phosphate 5-kinase. AB - In many different cell types, including smooth muscle cells (Baron et al., 1989, Am. J. Physiol., 256: C375-383; Baron et al., J. Pharmacol. Exp. Ther. 266: 8 15), phosphatidylinositol (4)-phosphate 5-kinase plays a critical role in the regulation of membrane concentrations of phosphatidylinositol (4,5)-bisphosphate and formation of inositol (1,4,5)-trisphosphate. In unstimulated porcine trachealis smooth muscle, 70% of total cellular phosphatidylinositol (4) phosphate 5-kinase activity was associated with cytoskeletal proteins and only trace activity was detectable in isolated sarcolemma. Using two different preparations, we studied cytoskeleton-associated phosphatidyl inositol (4) phosphate 5-kinase under conditions that attempted to mimic the ionic and thermal cytoplasmic environment of living cells. The cytoskeleton-associated enzyme, studied using phosphatidylinositol (4)-phosphate substrate concentrations that produced phosphatidylinositol 4,5-bisphosphate at about 10% of the maximal rate, was sensitive to free [Mg2+], had an absolute requirement for phosphatidylserine, phosphatidic acid, or phosphatidylinositol, and included type I isoforms. At 0.5 mM free [Mg2+], physiological spermine concentrations, 0.2-0.4 mM, increased phosphatidylinositol (4)-phosphate 5-kinase activity two to four times compared to controls run without spermine. The EC50 for spermine-evoked increases in activity was 0.17 +/- 0.02 mM. Spermine-evoked enzyme activity was a function of both free [Mg2+] and substrate concentration. Cytoskeleton-associated phosphatidylinositol (4)-phosphate 5-kinase was inhibited by free [Ca2+] over a physiological range for cytoplasm--10(-8) to 10(-5) M, an effect independent of the presence of calmodulin. Na+ over the range 20 to 50 mM also inhibited this enzyme activated by 5 mM Mg2+ but had no effect on spermine-activated enzyme. Na+, Ca2+, and spermine appear to be physiological modulators of smooth muscle cytoskeleton-bound phosphatidylinositol (4)-phosphate 5-kinase. PMID- 9731757 TI - Modulation of cellular tryptophan metabolism in human fibroblasts by transforming growth factor-beta: selective inhibition of indoleamine 2,3-dioxygenase and tryptophanyl-tRNA synthetase gene expression. AB - Alterations in the rate of cellular tryptophan metabolism are involved in mediating important biological activities associated with cytokines and growth factors. Indoleamine 2,3-dioxygenase (IDO) and tryptophanyl-tRNA synthetase are enzymes of tryptophan metabolism whose expression in a variety of cells and tissues is highly inducible by interferon-gamma (IFN-gamma). Transforming growth factor-beta (TGF-beta) antagonizes many cellular responses to IFN-gamma. The interaction of these two cytokines plays an important role in maintaining homeostasis during inflammation and repair. In human skin and synovial fibroblasts in vitro, TGF-beta caused time- and dose-dependent abrogation of IFN gamma-stimulated expression of IDO and tryptophanyl-tRNA synthetase mRNAs. The inhibition was selective and did not appear to be due to down-regulation of IFN gamma signaling by TGF-beta. In parallel with its effect on IDO mRNA expression, TGF-beta caused a marked reduction in intracellular IDO protein levels and abrogated IDO activity and tryptophan catabolism in these cells induced by IFN gamma. IFN-gamma caused a rapid and striking increase in the amount of IDO heterogeneous nuclear pre-mRNA and induced transcription of the IDO gene, as demonstrated by transient transfection assays. TGF-beta partially reversed this stimulation. IFN regulatory factor (IRF)-1 and stat1 are cellular intermediates in IFN signaling. Both are implicated in activation of IDO transcription in response to IFN-gamma. The stimulation by IFN-gamma of IRF-1 protein and mRNA expression was not prevented by treatment of fibroblasts with TGF-beta. Furthermore, gel mobility shift assays indicated that TGF-beta did not inhibit the induction of stat1 and IRF-1 binding activity to their cognate DNA recognition sites in the IDO gene promoter. In contrast, the stability of IDO mRNA transcripts was reduced in fibroblasts treated with TGF-beta, as shown by determination of mRNA half-lives following blockade of transcription with 5,6 dichlorobenzimidazole riboside. The findings indicate that TGF-beta prevents the induction of IDO and tryptophanyl-tRNA synthetase gene expression in fibroblasts. The repression of IDO expression by TGF-beta is mediated at both transcriptional and posttranscriptional levels. These results implicate TGF-beta in the negative regulation of tryptophan metabolism, provide evidence for the molecular basis of this regulation, and indicate that cellular tryptophan metabolism is under tight immunological control. PMID- 9731758 TI - Glycosaminoglycans bind granulocyte-macrophage colony-stimulating factor and modulate its mitogenic activity and signaling in human osteoblastic cells. AB - We recently demonstrated that granulocyte-macrophage colony-stimulating factor (GM-CSF) is an autocrine growth factor for human osteoblastic (hOB) cells. Since GM-CSF is a member of the heparin-binding factor family, we examined the interactions between GM-CSF and glycosaminoglycans (GAGs) present in the osteoblast microenvironment. Using a bioassay in which the mitogenic activity of recombinant human (rh) GM-CSF was measured after incubation in the presence of an hOB cell layer or extracellular matrix (ECM) produced by these cells, we showed that rhGM-CSF binds to GAG components present in the ECM and that the bound rhGM CSF retains its ability to stimulate hOB cell proliferation. Heparan sulfate compounds on the hOB cell surface were also found to sequester GM-CSF. Moreover, treatment with sodium chlorate, an inhibitor of GAG sulfation, suppressed the mitogenic activity of rhGM-CSF on hOB cells. This inhibitory effect was rescued by a low dose of heparin. Heparin was also found to promote the effect of rhGM CSF on hOB cell proliferation, allowing nonmitogenic high doses of rhGM-CSF to stimulate hOB cell growth. Western blot analysis showed that undersulfation of cellular GAGs by chlorate inhibited the increased tyrosine phosphorylation of proteins involved in GM-CSF signaling in cloned immortalized hOB cells. The data demonstrate that GM-CSF binds to proteoglycans on the hOB cell surface and in ECM produced by these cells and that the bound GM-CSF is biologically active. Furthermore, this study shows that cellular proteoglycans play an essential role in GM-CSF signaling and biological activity in hOBs. PMID- 9731759 TI - Love that beta-sheet. PMID- 9731760 TI - Fat sites found! PMID- 9731761 TI - New insights into sequence-dependent DNA structure. PMID- 9731762 TI - The energy landscape in non-biological and biological molecules. PMID- 9731763 TI - Chasing collagen. PMID- 9731764 TI - Picture story. Adding to the transcription rap sheet. PMID- 9731765 TI - Picture story. An RNA-RNA activator. PMID- 9731766 TI - OK, and the alpha-helix. PMID- 9731767 TI - Crystal structure of Escherichia coli HdeA. PMID- 9731769 TI - Solution structure of the tobramycin-RNA aptamer complex. AB - We have solved the solution structure of the aminoglycoside antibiotic tobramycin complexed with a stem-loop RNA aptamer. The 14 base loop of the RNA aptamer 'zippers up' alongside the attached stem through alignment of four mismatches and one Watson-Crick pair on complex formation. The tobramycin inserts into the deep groove centered about the mismatch pairs and is partially encapsulated between its floor and a looped out guanine base that flaps over the bound antibiotic. Several potential intermolecular hydrogen bonds between the charged NH3 groups of tobramycin and acceptor atoms on base pair edges and backbone phosphates anchor the aminoglycoside antibiotic within its sequence/structure specific RNA binding pocket. PMID- 9731768 TI - The synaptic SNARE complex is a parallel four-stranded helical bundle. AB - The heterotrimeric synaptic soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complex, consisting of the synaptic vesicle-associated membrane protein 2 (VAMP2) and presynaptic plasma membrane proteins SNAP-25 (synaptosome-associated protein of 25,000 Mr) and syntaxin 1A, is a critical component of the exocytotic machinery. We have used spin labeling electron paramagnetic resonance spectroscopy to investigate the structural organization of this complex, particularly the two predicted helical domains contributed by SNAP 25. Our results indicate that the N- and C-terminal domains of SNAP-25 are parallel to each other and to the C-terminal domain of syntaxin 1A. Based on these findings, we propose a parallel four-stranded coiled coil model for the structure of the synaptic SNARE complex. PMID- 9731770 TI - Evidence for an unfolding and refolding pathway in cytochrome c. AB - It has been suggested that three partially unfolded forms detected in a native state hydrogen exchange study of oxidized cytochrome c may represent sequential intermediates in an unfolding-refolding pathway. To better define the structure of each intermediate a 'stability labeling' method was used in which the stability of a given segment against unfolding was changed. The condition--folded or unfolded--of the stability-labeled segment in each intermediate could then be determined. The structures found are discrete and native-like and are just those necessary for a sequential pathway. PMID- 9731771 TI - Engineering an intertwined form of CD2 for stability and assembly. AB - The amino-terminal domain of CD2 has the remarkable ability to fold in two ways: either as a monomer or as an intertwined, metastable dimer. Here we show that it is possible to differentially stabilize either fold by engineering the CD2 sequence, mimicking random mutagenesis events that could occur during molecular evolution. Crystal structures of a hinge-deletion mutant, which is stable as an intertwined dimer, reveal domain rotations that enable the protein to further assemble to a tetramer. These results demonstrate that a variety of folds can be adopted by a single polypeptide sequence, and provide guidance for the design of proteins capable of further assembly. PMID- 9731772 TI - The crystal structure of the hyperthermophile chromosomal protein Sso7d bound to DNA. AB - Sso7d and Sac7d are two small (approximately 7,000 Mr), but abundant, chromosomal proteins from the hyperthermophilic archaeabacteria Sulfolobus solfataricus and S. acidocaldarius respectively. These proteins have high thermal, acid and chemical stability. They bind DNA without marked sequence preference and increase the Tm of DNA by approximately 40 degrees C. Sso7d in complex with GTAATTAC and GCGT(iU)CGC + GCGAACGC was crystallized in different crystal lattices and the crystal structures were solved at high resolution. Sso7d binds in the minor groove of DNA and causes a single-step sharp kink in DNA (approximately 60 degrees) by the intercalation of the hydrophobic side chains of Val 26 and Met 29. The intercalation sites are different in the two complexes. Observations of this novel DNA binding mode in three independent crystal lattices indicate that it is not a function of crystal packing. PMID- 9731773 TI - An atomic model of fimbrin binding to F-actin and its implications for filament crosslinking and regulation. AB - Using a new procedure that combines electron-density correlation with biochemical information, we have fitted the crystal structure of the N-terminal actin-binding domain of human T-fimbrin to helical reconstructions of fimbrin-decorated actin filaments. The map locates the N-terminal calcium-binding domain and identifies actin-binding site residues on the two calponin-homology domains of fimbrin. Based on this map, we propose a model of a fimbrin crosslink in an actin bundle and its regulation by calcium. PMID- 9731774 TI - Regulation of SNARE complex assembly by an N-terminal domain of the t-SNARE Sso1p. AB - The fusion of intracellular transport vesicles with their target membranes requires the assembly of SNARE proteins anchored in the apposed membranes. Here we use recombinant cytoplasmic domains of the yeast SNAREs involved in Golgi to plasma membrane trafficking to examine this assembly process in vitro. Binary complexes form between the target membrane SNAREs Sso1p and Sec9p; these binary complexes can subsequently bind to the vesicle SNARE Snc2p to form ternary complexes. Binary and ternary complex assembly are accompanied by large increases in alpha-helical structure, indicating that folding and complex formation are linked. Surprisingly, we find that binary complex formation is extremely slow, with a second-order rate constant of approximately 3 M(-1) s(-1). An N-terminal regulatory domain of Sso1p accounts for slow assembly, since in its absence complexes assemble 2,000-fold more rapidly. Once binary complexes form, ternary complex formation is rapid and is not affected by the presence of the regulatory domain. Our results imply that proteins that accelerate SNARE assembly in vivo act by relieving inhibition by this regulatory domain. PMID- 9731775 TI - Structure of the ATP-dependent oligomerization domain of N-ethylmaleimide sensitive factor complexed with ATP. AB - N-ethylmaleimide-sensitive factor (NSF) is a hexameric ATPase which primes and/or dissociates SNARE complexes involved in intracellular fusion events. Each NSF protomer contains three domains: an N-terminal domain required for SNARE binding and two ATPase domains, termed D1 and D2, with D2 being required for oligomerization. We have determined the 1.9 A crystal structure of the D2 domain of NSF complexed with ATP using multi-wavelength anomalous dispersion phasing. D2 consists of a nucleotide binding subdomain with a Rossmann fold and a C-terminal subdomain, which is structurally unique among nucleotide binding proteins. There are interactions between the ATP moiety and both the neighboring D2 protomer and the C-terminal subdomain that may be important for ATP-dependent oligomerization. Of particular importance are three well-ordered and conserved lysine residues that form ionic interactions with the beta- and gamma-phosphates, one of which likely contributes to the low hydrolytic activity of D2. PMID- 9731776 TI - Insights into transition state stabilization of the beta-1,4-glycosidase Cex by covalent intermediate accumulation in active site mutants. AB - The catalytic mechanism of 'retaining' beta-glycosidases has been the subject of considerable interest and debate for many years. The visualization of a covalent glycosyl enzyme intermediate by X-ray crystallography was first accomplished with a saccharide substrate substituted with fluorine at its 2-position. The structure implicated major roles for residue His 205 and for the 2-hydroxyl position of the proximal saccharide in binding and catalysis. Here we have studied the kinetic behavior of various His 205 mutants. One of these mutants, a double mutant H205N/E127A, has been used to stabilize a covalent glycosyl-enzyme intermediate involving an unsubstituted sugar, permitting crystallographic analysis of the interactions between its 2-hydroxyl group and the enzyme. PMID- 9731777 TI - Conserved mode of peptidomimetic inhibition and substrate recognition of human cytomegalovirus protease. AB - Human cytomegalovirus (HCMV) protease belongs to a new class of serine proteases, with a unique polypeptide backbone fold. The crystal structure of the protease in complex with a peptidomimetic inhibitor (based on the natural substrates and covering the P4 to P1' positions) has been determined at 2.7 A resolution. The inhibitor is bound in an extended conformation, forming an anti-parallel beta sheet with the protease. The P3 and P1 side chains are less accessible to solvent, whereas the P4 and P2 side chains are more exposed. The inhibitor binding mode shows significant similarity to those observed for peptidomimetic inhibitors or substrates of other classes of serine proteases (chymotrypsin and subtilisin). HCMV protease therefore represents example of convergent evolution. In addition, large conformational differences relative to the structure of the free enzyme are observed, which may be important for inhibitor binding. PMID- 9731778 TI - Crystal structure of human serum albumin complexed with fatty acid reveals an asymmetric distribution of binding sites. AB - Human serum albumin (HSA) is the most abundant protein in the circulatory system. Its principal function is to transport fatty acids, but it is also capable of binding a great variety of metabolites and drugs. Despite intensive efforts, the detailed structural basis of fatty acid binding to HSA has remained elusive. We have now determined the crystal structure of HSA complexed with five molecules of myristate at 2.5 A resolution. The fatty acid molecules bind in long, hydrophobic pockets capped by polar side chains, many of which are basic. These pockets are distributed asymmetrically throughout the HSA molecule, despite its symmetrical repeating domain structure. PMID- 9731779 TI - Ex vivo gene therapy prevents chronic graft vascular disease in cardiac allografts. AB - OBJECTIVE: We hypothesized that ex vivo hyperbaric transfection of antisense oligodeoxynucleotides for blockade of intercellular adhesion molecule-1, an important mediator of cell adhesion and T-cell co-stimulation, would reduce chronic graft vascular disease in cardiac allografts. METHODS: PVG hearts underwent ex vivo transfection with antisense, reverse antisense intercellular adhesion molecule-1 oligodeoxynucleotide (80 micromol/L), or saline solution at 3 atm pressure for 45 minutes at 4 degrees C and were transplanted heterotopically into ACI recipients with or without treatment with intercellular adhesion molecule-1 (1A29) or leukocyte function associated antigen-1 (WT.1) monoclonal antibodies. Transfection efficiency was confirmed with fluorescein isothiocyanate labeled oligodeoxynucleotides and fluorescent microscopy. Efficacy of intracellular adhesion molecule-1 blockade was assessed with the use of immunohistochemistry. Graft reperfusion injury was evaluated at 6 to 24 hours by neutrophil infiltration (myeloperoxidase [MPO]), cardiac edema (%wt/wt), and histologic injury (percent contraction band necrosis). Grafts from recipients treated with cyclosporine A (5 mg/kg per day, days 0 to 9) were scored for chronic graft vascular disease on postoperative day 90 ranging from 0 (no involvement) to 4 (>50% vascular occlusion). RESULTS: Transfection was highly efficient (fluorescein isothiocyanate-labeled oligodeoxynucleotides in 48%+/-5% of total myocardial nuclei) and effective at blocking intracellular adhesion molecule-1 expression (positive area in allografts taken on postoperative day 3 was reduced from 100%+/-0% to 52%+/-14%, n=4). Blockade with antisense oligodeoxynucleotides versus monoclonal antibodies was less effective at preventing reperfusion injury while more effective at reducing chronic graft vascular disease (score 0.98+/-0.48, p < 0.05). Reverse antisense oligodeoxynucleotides and vector control (antisense oligodeoxynucleotide infusion without pressure) groups failed to demonstrate this beneficial effect. CONCLUSION: Hyperbaric transfection of antisense oligodeoxynucleotides proved highly efficient, effective at blockade of intracellular adhesion molecule-1, and demonstrated a sequence-specific reduction in chronic graft vascular disease. This highly targeted alteration of donor organ immunogenicity may have an important future role in clinical immunosuppressive strategies. PMID- 9731780 TI - Tracheal transplantation for carinal reconstruction in dogs. AB - BACKGROUND: Experimental carinal allotransplantation has been performed with tracheocarinal Y-shaped allografts in dogs. In this study we tried canine carinal reconstruction with cylindrical allografts. MATERIAL AND METHODS: Carinal reconstruction was performed with allotransplantation of cylindrical trachea in dogs, and graft healing was evaluated by bronchoscopic observation, mucosal blood flow measurement, and histologic examination. A section of the recipient carina containing five tracheal rings and two main stem bronchi was removed, and a donor trachea seven rings long was inserted between the recipient trachea and the left main stem bronchus; then side-to-end anastomosis was performed between the graft midportion and recipient right main stem bronchus (new carina). The grafts were wrapped with pedicled omentum. Fresh grafts were transplanted into one group of dogs (n=8 ), and grafts cryopreserved for 1 week were transplanted into another group (n=7). RESULTS: No anastomotic leakage occurred in any dog. Excellent healing of grafts and graft anastomoses was observed by fiberoptic bronchoscopy in six dogs (75%) in the fresh graft group and in four dogs (57%) in the cryopreserved graft group. The mucosal blood flow in the new carina decreased remarkably and, although it recovered, mucosal blood flow remained under the preoperative level on day 28 after the operation. CONCLUSION: Cylindrical tracheal allotransplantation is useful for carinal reconstruction, and the method of side-to-end anastomosis between the donor trachea and recipient bronchus is a feasible and accessible procedure in dogs. PMID- 9731781 TI - Bronchoscopic treatment of intraluminal typical carcinoid: a pilot study. AB - OBJECTIVE: The curative potential of various bronchoscopic treatments such as Nd:YAG laser, photodynamic therapy, and brachytherapy for the treatment of intraluminal tumor has been reported previously. Bronchoscopic treatment can be used to treat small intraluminal tumor with curative intent, such as in patients with roentgenologically occult squamous cell cancer. In a retrospective study, we showed that bronchoscopic treatment provided excellent local control with surgical proof of cure in 6 of 11 patients with intraluminal typical bronchial carcinoid. METHODS: In a prospective study, 19 patients (8 women and 11 men) with resectable intraluminal typical bronchial carcinoid have undergone bronchoscopic treatment under general anesthesia. Median age was 44 years (range, 20-74 years). If tumor persisted after 2 bronchoscopic treatment sessions, surgery was performed within 4 months after the treatment. RESULTS: Bronchoscopic treatment was able to completely eradicate tumor in 14 of the 19 patients (complete response rate 73%, 95% CI: 49%-91%). Median follow-up of these patients is 29 months (range, 8-62 months). One patient had severe cicatricial stenosis after bronchoscopic treatment, and sleeve lobectomy was necessary. No residual carcinoid was found in the resected specimen. In the remaining 5 patients, bronchoscopic treatment did not result in a complete response and radical surgical resection was performed afterward with confirmation of residual carcinoid in the resected specimen. Median follow-up of the surgical group is 34 months (range, 12-62 months). CONCLUSIONS: Current data suggest that bronchoscopic treatment may be an effective alternative to surgical resection in a subgroup of patients with resectable intraluminal typical bronchial carcinoid. It alleviated the necessity of surgical resection in 68% (95% CI: 43%-87%) of the patients. PMID- 9731782 TI - Prognostic assessment of 1310 patients with non-small-cell lung cancer who underwent complete resection from 1980 to 1993. AB - OBJECTIVE: The TNM staging system of lung cancer is widely used as a guide for estimating prognosis and selecting treatment modality. In 1997, the International Union Against Cancer and the American Joint Committee on Cancer have adopted a revised stage grouping for lung cancer. However, the validity of the new stage grouping has not been fully established. We investigated the prognoses of patients who had resection of non-small-cell lung cancer to confirm the validity of the revised classification. METHODS: A total of 1310 patients with non-small cell lung cancer underwent complete resection and pathologic staging of the disease in our hospitals from 1980 through 1993. A pulmonary resection was performed with a systematic nodal dissection. The survivals were calculated with the Kaplan-Meier method on the basis of overall deaths, and the survival curves were compared by log rank test. RESULTS: There were significant differences in survival between patients with T1 N0 M0 and T2 N0 M0 disease and between those with T1 N1 M0 and T2 N1 M0 disease. However, there was no significant difference between patients with T2 NO M0 disease and those with T1 N1 M0 disease. No significant difference in survival was observed among patients with T2 N1 M0, T3 NO M0, and T3 N1 M0 cancer. Patients with different invaded organs of T3 subdivision (pleura, chest wall, pericardium, or diaphragm) had a different prognosis. There was no significant difference between patients with T3 N2 M0 disease and those with stage IIIB disease. CONCLUSIONS: We supported most of the revision, such as dividing stage I, dividing stage II, and putting T3 N0 M0 to stage IIB. Furthermore, we found some candidates for a subsequent revision, such as putting T3 N1 M0 to stage IIB, putting T2 N0 M0 and T1 N1 M0 together, regarding diaphragm invasion as T4, and putting T3 N2 M0 to stage IIIB. PMID- 9731784 TI - Intermediate survival in neonates with aortic atresia: a multi-institutional study. The Congenital Heart Surgeons Society. AB - OBJECTIVE: Controversy persists with regard to the treatment of patients with aortic atresia. Staged reconstructive operations and primary transplantation have been advocated as treatment strategies, but in many instances no treatment is undertaken. A multi-institutional study was undertaken for the purpose of characterizing this challenging patient group, comparing the prevalence and outcomes of the various treatment strategies, and identifying potential predictors of success or failure with each. METHODS AND RESULTS: A total of 323 neonates with aortic atresia were entered into a 21-institution prospective, nonrandomized study between January 1, 1994, and January 1, 1997. Three protocols were used, nonexclusively in many institutions: (1) staged reconstructive surgery with initial palliation by a Norwood procedure and eventual Fontan operation, (2) heart transplantation as initial definitive therapy, and (3) nonsurgical management. Analysis was based on initial protocol assignment: staged reconstructive surgery in 253 patients, heart transplantation in 49 patients, and nonsurgical management in 21 patients. For all patients initially entered into the 2 surgical treatment protocols, survival at 1, 3, 12, 24, and 36 months after entry was 67%, 59%, 52%, 51%, and 50%, respectively. A multivariable analysis found incremental risk factors for death at any time after entry to be lower birth weight (P=.04), associated noncardiac anomaly (P=.007), and entry into the nonsurgical protocol (P < .0001) or the staged reconstructive surgery protocol (P=.03). Four institutions had higher survival statistics; 2 used a heart transplantation protocol and 2 used a staged reconstructive surgery protocol. For the 113 patients treated at these 4 institutions, survival at 1, 3, 12, 24, and 36 months after entry was 77%, 70%, 64%, 62%, and 61%, respectively. Survival among the 4 institutions was similar (P=0.1). CONCLUSIONS: Among patients with aortic atresia, other features of cardiac structure including aortic size, degree of left ventricular hypoplasia, and degree of mitral hypoplasia or atresia are not predictive of survival from 2 surgical protocols. The highest survival was achieved with either treatment strategy at institutions strongly committed to the use of one or the other surgical management protocol. PMID- 9731783 TI - Serum carcinoembryonic antigen as an adjunct to preoperative staging of lung cancer. AB - OBJECTIVE: To determine whether measurement of preoperative serum carcinoembryonic antigen concentrations adds useful prognostic data to current preoperative staging of lung cancer by computed tomography, bronchoscopy, and mediastinoscopy. METHODS: A prospective cohort study of 130 consecutive patients was evaluated for suspected lung cancer from July 1991 through December 1992 at a university-affiliated Veterans Affairs Medical Center. Serum concentrations of carcinoembryonic antigen were measured before diagnosis, staging, or resection of cancer. RESULTS: Malignant disease was diagnosed by bronchoscopy, needle biopsy, mediastinoscopy, or resection in 111 of 130 patients. In the 50 patients undergoing resection with curative intent, multivariate analysis indicated that carcinoembryonic antigen was a significant predictor of survival independent of patient age, pathologic stage, histologic type, and tumor size (P=.0357). CONCLUSIONS: Elevated preoperative serum concentrations of carcinoembryonic antigen predict a poor prognosis for lung cancer independent of other conventional staging parameters and have an adjunctive role in the staging of lung cancer. PMID- 9731785 TI - Effects of various flow types on maternal hemodynamics during fetal bypass: is there nitric oxide release during pulsatile perfusion? AB - OBJECTIVE: This study investigates the role of various flow conditions on maternal hemodynamics during fetal cardiopulmonary bypass. METHODS: Normothermic fetal bypass was conducted under pulsatile, or steady flow, for a 60-minute period. Fetal lamb preparations were randomly assigned to 1 of the 3 groups: steady flow (n=7), pulsatile flow (n=7), or pulsatile blocked flow bypass (n=7), where fetuses were perfused with Nomega-nitro-L-arginine after the first 30 minutes of pulsatile flow to assess the potential role of endothelial autacoids. RESULTS: Maternal oximetry and pressures remained unchanged throughout the procedure. Under fetal pulsatile flow, maternal cardiac output increased after 20 minutes of bypass and remained significantly higher than under steady flow at minute 30 (8.8+/-0.7 L x min(-1) vs 5.9+/-0.5 L x min(-1), P=.02). Maternal cardiac output in the pulsatile group also remained higher than in both steady and pulsatile blocked flow groups, reaching respectively 8.7+/-0.9 L x min(-1) vs 5.8+/-0.4 L x min(-1) (P=.02) and 5.9+/-0.3 L min(-1) (P=.01) at minute 60. Maternal systemic vascular resistances were significantly lower under pulsatile than under steady flow after 30 minutes and until the end of bypass (respectively, 9.1+/-0.6 IU vs 12.7+/-1.1 IU, P=.02 and 8.9+/-0.5 IU vs 12.9+/ 1.2 IU, P=.01). Infusion of Nomega-nitro-L-arginine was followed by an increase in systemic vascular resistances from 9.3+/-0.7 IU, similar to that of the pulsatile group, to 13.5+/-1 IU at 60 minutes, similar to that of the steady flow group. CONCLUSIONS: Maternal hemodynamic changes observed under fetal pulsatile flow are counteracted after infusion of Nomega-nitro-L-arginine, suggesting nitric oxide release from the fetoplacental unit under pulsatile fetal flow conditions. PMID- 9731786 TI - Is return of angina after coronary artery bypass grafting immutable, can it be delayed, and is it important? AB - BACKGROUND: Because survival after either an operation or angioplasty is similar across a wide spectrum of coronary patients, lasting symptom relief assumes high priority. OBJECTIVES: The objectives of this observational clinical study were (1) to determine whether the return of angina is immutable; (2) to identify factors that might delay its return, and (3) to evaluate whether its return is predictive of subsequent adverse events. METHODS: The return of angina of any degree of severity and morbid events subsequent to its return were studied by multivariable time-related analyses in a consecutive series of 9600 patients who were undergoing primary isolated coronary bypass operations between 1971 and 1992. RESULTS: The freedom rate from return of angina was 94%, 82%, 61% and 38% at 1, 5, 10, and 15 years. Increased modest risk of early return of angina was associated with preoperative demographic, symptom, coronary and vascular disease variables but reduced by more extensive arterial grafting. The ever-increasing risk of late return of angina was associated with demographic, symptomatic, left ventricular function, and coronary disease variables and was related strongly to comorbidity but was weakly reduced by controllable surgical variables. After the return of angina, 10-year freedom rate from infarct and survival was 71% and 68% respectively. CONCLUSIONS: (1) The risk of angina return increases relentlessly after operation, so it is likely immutable. (2) Delay of late angina return by use of arterial grafting is clinically trivial; control of noncardiac comorbidity may be more effective. (3) Fortunately, the return of angina after coronary artery bypass grafting has minimal impact on survival and is not predictive of imminent infarct. PMID- 9731787 TI - Difference in acetylcholine-induced nitric oxide release of arterial and venous grafts in patients after coronary bypass operations. AB - OBJECTIVES: In vivo investigation of nitric oxide release in coronary bypass grafts has not been reported. We studied acetylcholine-induced nitric oxide release in vivo of coronary bypass grafts and vasomotor responses to acetylcholine of grafted coronary arteries in patients after coronary bypass grafting. METHODS: We examined 24 internal thoracic artery grafts and 16 saphenous vein grafts in 39 patients. The mean ages of the patients were 65 years for the arterial grafts and 68 years for the venous grafts. Nitric oxide was measured as the plasma nitrite level by the Griess reaction. Before and after intragraft acetylcholine infusion (5 microg), blood was sampled from the distal end of the graft, and angiograms were taken and analyzed by cine-densitometry. RESULTS: The plasma nitrite concentration after stimulation with acetylcholine compared with the control value was 134%+/-52% at 4 minutes (p=0.05) and 184%+/ 107% at 6 minutes (p=0.01) in the arterial grafts; in the venous grafts these values were 101%+/-24% at 4 minutes (p=0.96) and 108%+/-36% at 6 minutes (p=0.69). Low-dose acetylcholine dilated the coronary arteries supplied by arterial grafts by 6.3%+/-16.6% whereas coronary arteries supplied by venous grafts were reduced by 9.8%+/-11.8% in diameter and the vasoactive responses were different (p=0.01). CONCLUSIONS: In vivo internal thoracic artery grafts had more endothelium-derived nitric oxide release in response to acetylcholine than did saphenous vein grafts after coronary bypass grafting. PMID- 9731788 TI - Hematocrit value on intensive care unit entry influences the frequency of Q-wave myocardial infarction after coronary artery bypass grafting. The Institutions of the Multicenter Study of Perioperative Ischemia (McSPI) Research Group. AB - OBJECTIVES: No data exist regarding "the best" hematocrit value after coronary artery bypass graft surgery. Transfusion practice varies, because neither an optimal hematocrit value nor a uniform transfusion trigger criterion has been determined. METHODS: To investigate the optimal hematocrit value, we studied 2202 patients undergoing coronary bypass. The hematocrit value on entry into the intensive care unit (IHCT) was categorized into three groups: high (> or = 34%), medium (25% to 33%), and low (< or = 24%). Characteristics and adverse events (outcomes) were compared, and the effect of IHCT on the risk of myocardial infarction was determined by logistic regression. RESULTS: High IHCT (> or = 34%) was associated with an increased rate of myocardial infarction (8.3% vs 5.5% vs 3.6%; p < or = 0.03, high, medium vs low) and with more severe left ventricular dysfunction (11.7% vs 7.4% and 5.7%; p=0.006, high, medium vs low). Mortality rate increased with higher IHCT when all the high-risk subgroups were combined (8.6% vs 4.5% vs 3.2%; p < 0.001, high, medium vs low). By multivariate analysis, IHCT remained the most significant predictor of adverse outcomes (relative risk high vs low 2.22, 95% confidence interval: 1.04 to 4.76). No characteristic, event, medication, or transfusion therapy confounded the relationship between IHCT and outcome. CONCLUSION: High IHCT is associated with a higher rate of myocardial infarction and is an independent predictor of infarction. On the basis of the risk of myocardial infarction, there is no rationale for transfusion to an arbitrary level after coronary artery bypass grafting. PMID- 9731789 TI - Validation of a new intraoperative technique to evaluate load-independent indices of right ventricular performance in patients undergoing cardiac operations. AB - BACKGROUND: Assessment of right ventricular performance in the perioperative period is difficult because there is no generally accepted method of measuring right ventricular volume. We set out to determine whether conductance technology could provide a valuable technique for the investigation of intraoperative right ventricular function. METHODS AND RESULTS: Three validating studies were performed in 25 patients undergoing routine coronary revascularization. Study 1: The influence of conductance catheter position in the right ventricle was examined in 10 patients. Insertion of the conductance catheter through the outflow tract was associated with a larger gain constant and a smaller parallel conductance compared with insertion through the tricuspid valve. Study 2: The reproducibility of contractility measurements with the use of a conductance catheter was examined in 7 additional patients. Removal and reinsertion of the conductance catheter was not associated with any significant difference in right ventricular volume or contractile function. Study 3: Right ventricular performance before and after cardiopulmonary bypass was compared in 8 additional patients. There was a fall in the slope of the right ventricular preload recruitable stroke work from 15.6 (3.8) to 11.0 (5.1) mm Hg (P=.01) and an increase in the slope of the end-diastolic pressure-volume relations from 0.05 (0.02) to 0.11 (0.05) mm Hg/mL (P=.001). CONCLUSIONS: The conductance technique can be used to study perioperative changes in right ventricular performance. Insertion of the conductance catheter through the outflow tract provides stable and reproducible data. There is significant impairment of right ventricular contractility in the early postoperative period. PMID- 9731790 TI - Valve replacement with a stentless bioprosthesis: versatility of the porcine aortic root. AB - OBJECTIVE: Stentless valves convey important hemodynamic benefits but are used selectively depending on aortic root structure. The Freestyle valve (Medtronic, Inc, Minneapolis, Minn) is a versatile device that can be implanted by different methods depending on operating conditions. We aimed to demonstrate that a stentless valve could be used in every patient without increased risk of morbidity or mortality. We documented the effects of this valve on clinical outcome and left ventricular mechanics. METHODS: The Freestyle valve was implanted by the modified subcoronary method into 200 consecutive unselected patients who received a tissue valve in the aortic position and by root replacement in 2. Forty-three percent were older than 75 years. Forty percent underwent coronary bypass. Detailed clinical and echocardiographic follow-up (Food and Drug Administration protocol) was used out to 3 years. RESULTS: Mean ischemic time was 43+/-6 minutes for isolated aortic valve replacement and 63+/ 14 minutes with concomitant procedures. Thirty-day mortality was 6%, none of the deaths being valve related. Hemodynamic function improved progressively with falling valve gradients and increased effective orifice areas. Left ventricular mass fell within normal limits over 2 years, but at 3 years there was a non-valve related upswing. No instances of valve thrombosis, hemolysis, or paravalvular leak were noted. Less than 5% had mild to moderate aortic regurgitation. CONCLUSIONS: The Freestyle valve can be used in virtually every patient with aortic valve disease and provides superlative hemodynamic outcome. Hospital mortality and morbidity are similar to those reported for stented valves in an elderly population. PMID- 9731791 TI - Insulin stimulates pyruvate dehydrogenase and protects human ventricular cardiomyocytes from simulated ischemia. AB - Impaired myocardial metabolism after cardioplegic arrest results in persistent anaerobic lactate production. Insulin may protect the heart from ischemia and reperfusion by enhancing myocardial metabolic recovery. However, the stimulation of glycolysis during ischemia may be detrimental because of an accumulation of metabolic end-products. We examined the effect of insulin on quiescent human ventricular cardiomyocytes subjected to simulated cardioplegic ischemia and reperfusion. METHODS: Primary cardiomyocyte cultures were established from patients undergoing corrective repair of tetralogy of Fallot. Cells were exposed to varying concentrations of glucose and insulin during 30 minutes of stabilization in 10 mL of phosphate-buffered saline solution. Ischemia was simulated by exposing the cells to a low volume (1.5 mL) of deoxygenated phosphate-buffered saline solution for 90 minutes followed by 30 minutes of simulated reperfusion in 10 mL of normoxic phosphate-buffered saline solution. Cell viability was assessed by trypan blue exclusion. The activity of mitochondrial pyruvate dehydrogenase was measured in 3 states: stabilization, ischemia, and reperfusion. In addition intracellular lactate, adenine nucleotides, extracellular lactate, pyruvate, and acid release were measured. RESULTS: Higher ambient glucose concentrations resulted in greater cellular injury although insulin-treated cells displayed less injury after ischemia and reperfusion. Insulin increased the pyruvate dehydrogenase activity by 31% in cardiomyocytes and reduced extracellular lactate production by 40%. Intracellular adenosine triphosphate was improved by 75% in cells exposed to high glucose concentrations in the presence of insulin. CONCLUSIONS: Insulin protected human ventricular cardiomyocytes from ischemia and reperfusion. This protection may be due to a stimulation of pyruvate dehydrogenase activity which resulted in improved aerobic metabolism. PMID- 9731792 TI - Isolated left ventricular myocyte contractility in patients undergoing cardiac operations. AB - BACKGROUND: Because of methods required for obtaining isolated left ventricular myocytes, evaluation of the contractile function of isolated left ventricular myocytes in normal human patients has been limited. Accordingly, the goal of the present study was to develop a means to isolate human left ventricular myocytes from small myocardial biopsy specimens collected from patients undergoing elective coronary artery bypass operations and to characterize indices of myocyte contractile performance. METHODS: Myocardial biopsy specimens were obtained from the anterior left ventricular free wall of 22 patients undergoing coronary artery bypass operations. Myocytes were isolated from these myocardial samples by means of a stepwise enzymatic digestion method and micro-trituration techniques. Isolated left ventricular myocyte contractile function was assessed by computer assisted high-speed videomicroscopy under basal conditions and in response to beta-adrenergic receptor stimulation with isoproterenol. RESULTS: A total of 804 viable left ventricular myocytes were successfully examined from all of the myocardial biopsy specimens with an average of 37+/-4 myocytes per patient. All myocytes contracted homogeneously at a field stimulation of 1 Hz with an average percent shortening of 3.7%+/-0.1% and shortening velocity of 51.3+/-1.3 microm/s. After beta-adrenergic receptor stimulation with isoproterenol, percent shortening and shortening velocity increased 149% and 118% above baseline, respectively (P < .05). CONCLUSION: The unique results of the present study demonstrated that a high yield of myocytes could be obtained from human left ventricular biopsy specimens taken during cardiac operations. These myocytes exhibited stable contractile performance and maintained the capacity to respond to an inotropic stimulus. The methods described herein provide a basis by which future studies could investigate intrinsic and extrinsic influences on left ventricular myocyte contractility in human beings. PMID- 9731793 TI - Regional cerebral blood flow during rewarming of cardiopulmonary bypass correlates with posthypothermic regional glucose use. AB - BACKGROUND AND OBJECTIVES: Although global measurements of cerebral blood flow and metabolism during and after profoundly hypothermic cardiopulmonary bypass have been performed both in experimental animals and in human beings, little is known about their regional changes. The purpose of this study was to investigate the changes in regional cerebral blood flow during profoundly hypothermic cardiopulmonary bypass and regional cerebral glucose use after cardiopulmonary bypass. METHODS: We measured regional cerebral blood flow with positron emission tomography during both the cooling (n=5) and rewarming (n=5) of hypothermic cardiopulmonary bypass in anesthetized dogs by continuously infusing 15O-labeled water. We altered the core temperature between 20 degrees and 37 degrees C. To assess the integrity of brain metabolism, we measured the regional cerebral glucose use by bolus injections of 18F-labeled 2-fluoro-2-deoxy-D-glucose. RESULTS: Regional cerebral blood flow decreased homogeneously during cooling. The regional cerebral blood flow at 20 degrees C was about one fourth of that at 37 degrees C. In contrast, at 24 degrees, 28 degrees , and 32 degrees C during rewarming, there were significant interregional differences in the regional cerebral blood flow for given temperatures (p=0.0075, 0.034, and 0.048, respectively). These interregional differences disappeared after rewarming. Although the regional cerebral blood flow significantly correlated with the regional cerebral glucose use in the control condition at 37 degrees C without cardiopulmonary bypass (r=0.75; p=0.00012), this correlation disappeared after profoundly hypothermic cardiopulmonary bypass (r=0.204; p=0.388). Regional cerebral blood flow at 32 degrees C during rewarming positively correlated with the regional cerebral glucose use after cardiopulmonary bypass (r=0.655; p=0.0017). CONCLUSION: The altered regional cerebral blood flow during rewarming of profoundly hypothermic cardiopulmonary bypass might affect regional brain metabolism. PMID- 9731794 TI - Interleukin-6 derived from hypoxic myocytes promotes neutrophil-mediated reperfusion injury in myocardium. AB - BACKGROUND: Reperfusion injury in the myocardium has recently been considered to be a type of inflammation, and close attention has been paid to the possible involvement of neutrophils, complement, and cytokines in the onset of this injury. Recently, it has been reported that serum levels of interleukin-6 are elevated significantly after myocardial infarction. The major site of interleukin 6 production and its exact roles are still unknown. In this study, we hypothesized that myocytes may produce interleukin-6 during hypoxia and this may play a role in neutrophil-mediated reperfusion injury. METHODS AND RESULTS: In the clinical study, 20 patients who underwent coronary artery bypass grafting were divided into 2 groups: group F, in which patients were treated with a serine protease inhibitor (FUT-175, 2 mg/kg per hour) during cardiopulmonary bypass, and group C (untreated patients). In group C, myocardial interleukin-6 production, as determined by the difference between the interleukin-6 level in the cardiopulmonary bypass circuit and its level in coronary venous blood, increased significantly after reperfusion (12+/-4 pg/mL) as compared with that before aortic crossclamping (2+/-2 pg/mL). In group F, the increase in the interleukin-6 level was suppressed significantly (before aortic crossclamping, 3+/-2 pg/mL; after reperfusion, 4+/-3 pg/mL). The interleukin-6 production differed significantly between group C and group F. In the in vitro experimental study, the supernatant from myocytes exposed to 2 hours of hypoxia (group 2H) showed significantly higher levels of interleukin-6 (455+/-260 pg/mL) than that from normoxic myocytes (group N) (47+/-15 pg/mL). This interleukin-6 production was suppressed by the addition of FUT-175 (123+/-24 pg/mL). The interleukin-6 production by endothelial cells of coronary vessels did not differ between group 2H (283+/-151 pg/mL) and group N (151+/-86 pg/mL). In a coincubation system with a monolayer of endothelial cells on collagen membrane and myocytes under collagen membrane in a modified Boyden chamber, 2 hours of coincubation showed a significantly higher percent of neutrophil transendothelial migration (group 2H vs N, 78%+/-13% vs 26%+/-11%), value of chemiluminescence (22+/-8 vs 5+/-2 x 10(3) counts/3 minutes), and percent of irreversibly damaged myocytes (48%+/-17% vs 12%+/-8%) than normoxic coincubation. In contrast, anti-interleukin-6 monoclonal antibody significantly attenuated neutrophil transendothelial migration (42%+/-19%) and irreversible damage of myocytes (26%+/-15%) in 2 hours of coincubation. CONCLUSIONS: Interleukin-6 is produced from myocardium during ischemia and reperfusion in patients undergoing coronary bypass grafting. This interleukin-6 may be derived from hypoxic myocytes and play a role in neutrophil mediated reperfusion injury in myocardium. PMID- 9731795 TI - Postintubation tracheoesophageal fistula: surgical treatment of three cases. PMID- 9731796 TI - Surgical correction of the hypoplastic aortic arch by the subclavian free flap method in the neonate. PMID- 9731797 TI - Malignant fibrous histiocytoma of the heart producing interleukin-6. PMID- 9731798 TI - Left thoracodorsal artery as an inflow graft for minimally invasive direct coronary artery bypass grafting. PMID- 9731799 TI - Improved expandable prosthesis in postpneumonectomy syndrome with deformed thorax. PMID- 9731800 TI - Minimally invasive left anterior descending coronary artery bypass with right gastroepiploic artery graft. PMID- 9731801 TI - The relation between pump flow rate and pulsatility on cerebral hemodynamics during pediatric cardiopulmonary bypass. PMID- 9731802 TI - Aortic valves are antigenic but less so than myocardium. PMID- 9731803 TI - Repair of subdivided left atrium associated with persistent left superior vena cava. PMID- 9731804 TI - Early extubation after cardiac operations in neonates and young infants. PMID- 9731805 TI - Lesser sac endoscopy and laparoscopy in pancreatic carcinoma definitive diagnosis, staging and palliation. AB - Laparoscopy with lesser sac endoscopy (LSE) were used in combination from 1987 to 1992 in 103 patients for differentiation between pancreatic carcinoma and other peripancreatic pathology, staging, and palliation. LSE identified pancreatic carcinoma in 38 patients; pancreatic cystadenocarcinoma in 2 patients; pancreatic cystadenoma in 3 patients; pancreatic adenoma in 1 patient; pancreatic metastases from liver in 2 patients; and pancreatic cysts in 5 patients. False negative diagnosis of pancreatic carcinoma occurred in two cases. Nontumor pancreatic pathology was revealed in 10 patients. Specifically, acute pancreatitis was found in four patients, and chronic pancreatitis was found in six patients. Extrapancreatic cancers were identified in 15 patients: retroperitoneal extraorgan tumors were found in 2 patients; extrahepatic biliary tract cancer in 6 patients; gallbladder cancer in 1 patient; liver cancer in 3 patients; and stomach cancer in 1 patient. In five cases no pathology was found. Overall correct definitive diagnosis was established in 101 patients. Sensitivity of laparoscopy with LSE for pancreatic carcinoma diagnosis proved to be 95 per cent (38 of 40 patients), for pancreatic tumors diagnosis 96.22 per cent (51 of 53 patients); specificity of the method 100 per cent; and accuracy of diagnosis 98 per cent (101 of 103 patients). Thus, the accuracy of the method was as high as the accuracy of combination of all known modalities. Criteria of unresectability were revealed with the combination of LSE and laparoscopy in 75 per cent (30 of 40 cases) of pancreatic carcinoma. Moreover, laparoscopy allowed palliation of pancreatic carcinoma. Laparoscopic cholecystostomy was performed in 10 patients, and laparoscopic cholecystojejunostomy with enteroenterostomy was performed in 6 patients. PMID- 9731806 TI - Does lining polypropylene with polyglactin mesh reduce intraperitoneal adhesions? AB - A method that appears to reduce the rate of adhesion formation between intraperitoneal viscera and prosthetic mesh is the placement of absorbable mesh between nonabsorbable mesh and intraperitoneal viscera. In this study, polyglactin mesh was compared with nonabsorbable polypropylene mesh (Marlex). Forty-seven Sprague-Dawley rats were divided into four groups: 1) control, 2) polyglactin (Vicryl), 3) polypropylene mesh, and 4) polyglactin-lined polypropylene mesh. All rats that underwent mesh placement had midline laparotomy with anastamosis of mesh to fascial borders. Controls underwent midline laparotomy and closure only. Groups were then studied at 1, 2, and 3 months, respectively, to determine the degree of adhesion formation. Gross inspection was performed by a blinded researcher with numerical rank given based on the number of adhesions observed: 0, none; 1, mild; 2, moderate; and 3, severe. The data showed that rats in group 3 (polypropylene only) had significant adhesions at 3 months, with average numerical score of 2.75. Polyglactin and polyglactin/polypropylene groups had similar scores of 1.5 each. Control groups predictably showed little adhesion formation, with average score of 0.25. Based on these data, it is observed that lining polypropylene mesh with absorbable polyglactin mesh can reduce adhesion formation to nonabsorbable mesh. The difference in degree of adhesions is most notable at 3 months. This technique may be an important adjunct to reduce the clinical sequelae of intraperitoneal adhesions. PMID- 9731807 TI - Primary anastomosis in the treatment of acute disease of the unprepared left colon. AB - Between June 1, 1990 and December 31, 1996, 58 consecutive patients with unprepared colons were urgently explored for nontraumatic disease with intent to proceed with primary left-sided colonic anastomosis. Unprotected anastomoses were not attempted in 15 patients. The causes of exclusion included preoperative and intraoperative shock in three patients, and three patients were on long-term high dose steroids, four had gross fecal contamination of the peritoneal cavity, four had large pelvic abscesses, and one had ischemic colitis. All 43 patients undergoing anastomosis without protective colostomy had stapled anastomoses. Indications included complicated diverticular disease in 32 cases. There were nine cases of obstruction from colorectal carcinoma and one obstruction due to sigmoid volvulus. There was one case of perforation from pseudomembranous enterocolitis. The most common complications were: atelectasis in nine cases, wound infection in two cases, and prolonged ileus in two cases. Pelvic abscess occurred in one case. There was one wound dehiscence. There was one anastomotic dehiscence, and there was no mortality. Operative time averaged 85 minutes and hospital length of stay 9.7 days. Primary anastomosis of the unprepared left colon is safe in most urgent and emergent situations, thus avoiding the significant morbidity and cost of colostomy closure. PMID- 9731808 TI - Laparoscopic cholecystectomy in octogenarians. AB - Performance of laparoscopic cholecystectomy (LC) is increasing, and patients age 80 and over comprise an increasingly larger proportion of the LC population. This study documents that the increase is accompanied by safe outcome in this patient population. However, the evidence also suggests that cholelithiasis appears to have been a neglected condition in this age group. The prevalence of nonelective procedures, the conversion rate to an open operation, more intraoperative complications, and the percentage having evidence of common bile duct stone passage all support this assertion. With the technology of LC, we are now appropriately addressing the problem with a treatment that allows less surgical trauma to the patient and shorter recovery time. Same-day LC surgery for the octogenarian appears to be very safe and would justify a decision to perform earlier LC in these patients. Surgery done before the appearance of comorbid conditions that increase the surgical and anesthetic risks may result in improved outcomes for the elderly at lower cost. Even when necessary in the already hospitalized patient, LC can be accomplished with morbidity and mortality comparable to those of elective abdominal procedures in younger populations. PMID- 9731809 TI - Iliac artery ischemic: analysis of risks for ischemic complications. AB - Risk factors for lower extremity ischemic complications (ICs) following iliac arterial injuries have not been addressed. Patients with penetrating iliac artery injuries over a 15-year period were reviewed. IC was defined as compartment syndrome with or without tissue loss. Patients with iliac artery repair who developed ICs were compared with those without ICs (excluding early deaths from hemorrhagic shock). Comparison included demographics, severity of shock, physical examination, and operative findings. There were 94 arterial injuries in 80 patients (34 common iliac, 23 internal iliac, and 37 external iliac). There were 26 deaths (33%), and 3 patients were excluded for technical reasons. Of the 51 who underwent arterial reconstruction, 34 had no ischemia, whereas 17 (33%) had ICs (9 with tissue loss and 8 with compartment syndrome only). Immediate fasciotomies were performed in 6 of the IC patients due to the early recognition of compartment syndrome; 1 required amputation from the profound ischemia. Delayed recognition of compartment syndrome in the remaining 11 IC patients resulted in eight amputations (P < 0.05). We conclude that ICs following iliac arterial injuries significantly correlate with shock as indicated by systemic pH, lactate and transfusion requirements, and a preoperative pulseless extremity. In these patients, close monitoring of compartment pressures is necessary, and immediate fasciotomies should be strongly considered. PMID- 9731810 TI - Pancreatic injuries resulting from penetrating trauma: a multi-institution review. AB - Pancreatic injury from penetrating trauma continues to be a source of significant morbidity and mortality, with questions remaining regarding optimal treatment of injuries. Our goal was to evaluate current trends in the operative management of these injuries. Our patient population comprised all patients admitted to one of three Level I trauma centers over an 8-year period that had sustained penetrating pancreatic trauma. The study was a retrospective chart review. Sixty-two patients were identified. All had associated abdominal injuries, with the liver and stomach being the most commonly injured organs. There were 14 deaths (mortality 22.6%), 10 within the first 48 hours due to associated vascular injury. In the 52 patients surviving beyond 48 hours, there were 19 patients with injuries to the main pancreatic duct and 33 with parenchymal injuries only. Pancreatic resection was carried out for all patients with ductal injury except for one, who later required distal pancreatectomy for pseudocyst and pancreatic fistula. Significant pancreatic fistulae developed in five patients, three in patients treated by drainage and two in patients treated by resection. The incidence of fistula formation was significantly higher for drainage versus resection in the patients with ductal injuries. The incidences of other complications were not affected by type of pancreatic injury, associated injuries, or method of management. We conclude that the majority of deaths in patients with penetrating pancreatic trauma are due to associated organ or vascular injuries. Appropriate management of the pancreatic injury can reduce the long-term complications. These results support treating patients with suspected ductal injuries by appropriate resection. Drainage should probably be sufficient for most nonductal pancreatic injuries. PMID- 9731811 TI - Surgical clips: a cause of late recurrent gallstones. AB - The formation of gallstones around surgical clips after cholecystectomy is a rare complication, with only seven reported cases in the English literature since its initial description in 1979. Three other cases report clip migration into the common bile duct and obstruction. We report a recent experience with "clip cholelithiasis." A 78-year-old female, 16 years following cholecystectomy, presented with a several-month history of colicky abdominal pain worsened by meals, and a 1 week history of jaundice, anorexia, nausea, and vomiting. An abdominal ultrasound demonstrated dilatation of the biliary tree without visible choledocholithiasis. Endoscopic retrograde cholangiopancreatography demonstrated a 1.5-cm radiolucent stone in the common bile duct containing a central surgical clip. She was successfully treated with endoscopic sphincterotomy and stone retrieval. The first report of clip cholelithiasis occurred in 1979. Six additional cases have been reported as well as three cases of clip migration without stone formation into the common bile duct. The incidence of clip cholelithiasis may increase in frequency with the increased use of metallic clips during laparoscopic cholecystectomy. The occurrence of cholelithiasis around inert metals is rare and may be prevented using absorbable clips; however, stone formation is also reported around absorbable materials. PMID- 9731812 TI - Colorectal cancer in the young patient. AB - Although predominantly a disease of older adults, colorectal cancer affects the younger population with an incidence of two to six per cent. It is thought to carry a less favorable prognosis in the young than in the general population. This may be due to advanced stage of the tumor at diagnosis. This study is composed of 37 patients, aged 40 and younger, treated over a 20-year period for colorectal cancer at Louisiana State University Medical Center-Shreveport and E. A. Conway Hospital. It was performed to investigate the incidence, stage at diagnosis, and prognosis of colorectal cancer in these young patients. The location of the primary tumor was fairly evenly distributed throughout the colon and rectum in this population. Pain, weight loss, rectal bleeding, and nausea and vomiting were the most common presenting symptoms. A family history of colon cancer or premalignant lesions were not risk factors in this study. Seventy per cent of all patients were treated with curative intent, and 42 per cent of these patients developed recurrent disease. The patients in this review presented with a higher incidence of advanced disease. Thirty-seven per cent of the lesions were Duke's C and 22 per cent were Duke's D, with poor 5-year survival (11% and 0%, respectively) when compared with national studies. The absolute 5-year survival for all young patients with colorectal cancer was 26 per cent (5 of 19 patients). It is important for the surgeon to be aware of the potential for colorectal cancer in young patients and to take an aggressive approach to the diagnosis and early treatment of the disease. PMID- 9731814 TI - Late follow-up and functional outcome after traumatic reproductive tract injuries in women. AB - The aim of this study was to assess female reproductive tract injuries and late effects on sexual and reproductive function. This was a review of women presenting to a Level I trauma center with reproductive tract injuries over 12 years. Thirty-one women (average age, 30 years) were divided into coital (19) and noncoital (12) injury groups. One-third of coital trauma presented late, one fourth was abuse related, and seven women presented in shock. All had vaginal lacerations, and 15 required repair. Follow-up in 6 of 19 (32%) women averaged 4.5 years. Noncoital injuries resulted primarily from vehicular trauma, and two thirds had associated abdominal injuries. Interventions included: vaginal laceration irrigation/repair (4), salpingectomy (2), ovariectomy (2), repair uterine perforation (1), and emergency cesarean section (2). The average Injury Severity Score was 25, with two deaths. Follow-up in 6 of 10 (60%) survivors averaged 6.1 years. The combined group has had seven subsequent pregnancies, and two women have minor dyspareunia after pelvic fracture. Women with coital injuries may develop shock, requiring rapid resuscitation and operative repair. Noncoital injuries are often associated with multiple severe injuries and require operative intervention. Late sequelae are minimal in both groups, and even severe injuries do not preclude normal pregnancy and sexual function. PMID- 9731813 TI - Covering the "open abdomen": a better technique. AB - "Damage control" in severe abdominal trauma, abdominal compartment syndrome, necrotizing fasciitis of the abdominal wall, and necrotizing pancreatitis often preclude closure of the fascia after laparotomy. Many techniques have been reported for temporary coverage of the exposed viscera, but most have had documented problems. We report the successful use, since 1989, of a temporary sutureless coverage. The viscera are covered with omentum when possible, then with a clear plastic sheet. Sump drains are placed over this layer. The entire abdomen is then covered with two layers of iodophor-impregnated adhesive plastic drape. The last 50 patients managed with this technique are reported. The most common indication (27 patients) was for treatment of severe abdominal trauma. There were no wound infections, fasciitis, or bowel obstruction. Eighteen patients died; no deaths were related to abdominal closure. Temporary abdominal covering with adhesive plastic sheeting is a rapid, safe, and readily available method for managing the open abdomen. This technique provides a physiologic milieu for the abdominal viscera, simplifies nursing care, and promotes safe closure of the abdomen at a later time. PMID- 9731815 TI - A protocol for the initial management of unstable pelvic fractures. AB - The initial management of life-threatening hemorrhage associated with severe pelvic fractures has long been a source of debate. A review of the literature reveals that many advocate emergent orthopedic external fixation (EX-FIX) for severe pelvic fractures, whereas others claim greater success with angiographic embolization (ANGIO) as the first line of treatment. Although many have attempted to classify management options by fracture pattern, to date there has been no prospective trial comparing outcomes for each method of treatment. We offer a prospective study of all pelvic fracture patients admitted to our Level I trauma center between July 1994 and July 1995. Patients were classified according to fracture pattern and degree of hemodynamic instability. Those with primarily anterior pelvic ring fractures underwent emergent EX-FIX for control of hemorrhage, whereas those with primarily posterior pelvic ring fractures underwent emergent ANGIO to control hemorrhage. We found that blood product requirements and hospital stay were similar in each group. However, the complication rate was higher in patients who underwent initial emergency EX-FIX, primarily because of failure to adequately control hemorrhage. We conclude that patients with anterior-posterior compression type 2 and 3, lateral compression type 2 and 3, or vertical shear injuries, who are hemodynamically unstable as a result of their pelvic fracture, should undergo immediate ANGIO if laparotomy is not indicated. If laparotomy is indicated, EX-FIX should be placed intraoperatively, followed by postoperative ANGIO. PMID- 9731816 TI - Use of an artificial neural network to predict length of stay in acute pancreatitis. AB - Length of stay (LOS) predictions in acute pancreatitis could be used to stratify patients with severe acute pancreatitis, make treatment and resource allocation decisions, and for quality assurance. Artificial neural networks have been used to predict LOS in other conditions but not acute pancreatitis. The hypothesis of this study was that a neural network could predict LOS in patients with acute pancreatitis. The medical records of 195 patients admitted with acute pancreatitis were reviewed. A backpropagation neural network was developed to predict LOS >7 days. The network was trained on 156 randomly selected cases and tested on the remaining 39 cases. The neural network had the highest sensitivity (75%) for predicting LOS >7 days. Ranson criteria had the highest specificity (94%) for making this prediction. All methods incorrectly predicted LOS in two patients with severe acute pancreatitis who died early in their hospital course. An artificial neural network can predict LOS >7 days. The network and traditional prognostic indices were least accurate for predicting LOS in patients with severe acute pancreatitis who died early in their hospital course. The neural network has the advantage of making this prediction using admission data. PMID- 9731817 TI - Management of bronchobiliary fistula as a late complication of hepatic resection. AB - Bronchobiliary fistula is an uncommon but remarkable complication after hepatic resection. The case reported illustrates the clinical presentation and preferred initial management of these fistulae. A 61-year-old white male underwent two wedge resections for colorectal metastases to the liver with removal of a portion of the right diaphragm. Four years later, he developed obstructive jaundice secondary to tumor recurrence in the porta hepatis, which required endoscopic stent placement, radiation, and chemotherapy. Almost 2 years later, he developed frank biliptysis. Percutaneous transhepatic cholangiography (PTC) revealed occlusion of the common hepatic duct stent and a bronchobiliary fistula. With adequate reestablishment of common duct drainage, the patient rapidly improved and was discharged free of symptoms. Bronchobiliary fistulae are rare complications of hepatic resection that can present from days to years after operation. Endoscopic retrograde cholangiopancreatography and PTC are the diagnostic studies of choice and offer the possibility of therapeutic intervention. Although large series in the literature emphasize the surgical management of bronchobiliary fistulae, the reoperative procedures tend to be complicated, with a significant morbidity and mortality. Nonsurgical interventions via endoscopic retrograde cholangiopancreatography or PTC are more recently notably successful when resolution of a distal biliary obstruction is accomplished. Only after aggressive attempts at nonoperative, interventional techniques have failed should operative approaches be entertained. PMID- 9731818 TI - The Internet and education in surgery. AB - The purpose of this review is to explain the developing role of the Internet and the World Wide Web (WWW) in promoting education in surgery. Internet sites relevant to surgery are appearing rapidly. Remote literature searches can query for surgery trials and results. Societies are using the WWW for transmission and review of publication materials. News groups interactively discuss current developments and trends. Surgeons are using personal and institutional sites to advertise services. Conventional slide shows migrate to the WWW for convenient downloading for surgeons and patients. Multimedia capabilities of the WWW expand the depth of information transmission, enabling education emanating from remote sites with narration and video depiction of procedures. These sophisticated tools can be demonstrated today with real online applications. One site facilitates surgical education using the WWW for program information, symposium coordination, links to regional subspecialty societies, residency cataloging, patient question and answer forums, and multimedia procedure descriptions. The principles of WWW communication used in this website can adapt to meet any educational need. The specialty of surgery is well suited to incorporation of online multimedia education over the Internet to follow new developments in our field. PMID- 9731819 TI - Localized malignant mesothelioma: a case report. AB - Localized malignant mesothelioma is an extremely rare form of presentation of malignant mesotheliomas. Only six cases have been reported. A 66-year-old male, former smoker, with occupational exposure to asbestos for 35 years, presented complaining of increasing fatigability, low-grade fever and anorexia for 4 weeks. The chest radiography showed a left lung mass. The computed tomography showed a 5 cm left posterior mass. The biopsy showed malignant cells. The patient underwent a surgical en bloc resection of the tumor. Pathology revealed a localized, poorly differentiated mesothelioma. Immunohistochemistry was positive for cytokeratin and vimentin while negative for carcinoembryonic antigen and Leu-M1. The final diagnosis was localized malignant mesothelioma. Aggressive resection of the tumor has shown to increase survival in previous reports, although the biological behavior of such tumors is still difficult to predict. PMID- 9731820 TI - Intrathyroid cysts of thyroglossal duct origin. AB - Thyroglossal duct cysts develop from a persistent portion of the thyroglossal tract and have been described as occurring anywhere from the base of the tongue to the manubrium. We present two patients who presented with a cystic thyroid nodule due to an intrathyroid thyroglossal duct cyst. A fine-needle aspiration biopsy was performed, which revealed benign squamous cells. Upon exploration, the first patient was found to have a 2-cm cyst within the thyroid isthmus, and in the second patient, a 1-cm cyst was found in the right thyroid lobe. Pathologic analysis revealed the cyst to be lined by a squamous epithelium consistent with a thyroglossal duct cyst. The lesions were completely surrounded by normal thyroid tissue. There was no evidence of a remnant of the thyroglossal duct extending from the thyroid in the region of the cyst. Both patients were treated by thyroid lobectomy and isthmusectomy and have remained without evidence of recurrence. Intrathyroid thyroglossal duct cysts should be included in the differential of patients with cystic thyroid lesions. Fine-needle aspiration revealing benign squamous cells is usually diagnostic and may detect an occult carcinoma arising within the cyst. Surgical resection is curative and should include a Sistrunk procedure if a thyroglossal duct tract is present. PMID- 9731821 TI - Retrosternal goiter: a six-year institutional review. AB - Retrosternal goiter is defined as any goiter in which at least 50 per cent of the thyroid resides below the level of the thoracic inlet. The incidence of retrosternal goiter varies from 3 to 20 per cent with respect to thyroidectomy patients. A retrospective chart review from June 1991 to December 1997 found 232 thyroidectomies performed at our institution. Sixteen patients were found to have retrosternal goiters (6.9%). The mean age was 57.8 years (range, 34-92). All were of benign pathology. Symptoms included shortness of breath (68.8%), hoarseness (37.5%), dysphagia (31.3%), and superior vena cava obstruction (6.25%). Thirteen patients were female (81.3%). Fifteen patients had surgical intervention (93.8%). Total thyroidectomy was performed in nine cases (60%), whereas lobectomy was performed in six cases (40%). All treated patients had complete resolution of symptoms. A cervical incision alone was used in 13 cases (86.7%). Complications consisted of one postoperative pleural effusion and in one case a traumatic C5 nerve root compression occurred. There were no instances of long-term vocal cord paralysis or hypoparathyroidism. There was no perioperative mortality. In the majority of patients with retrosternal goiter, surgery can be done expeditiously through a cervical incision with minimal morbidity and mortality. PMID- 9731822 TI - Interventions to reduce decibel levels on patient care units. AB - The University of Virginia Health System inpatient satisfaction survey identified noise as the most important irritant to surgical inpatients. Analysis of the level and pattern of noise on patient floors and intensive care units was done with baseline measurements followed by then two separate interventions: 1) education of nursing and physician staff 2) closing patient room doors. A decibel meter (M-27 Dosimeter) recorded the noise level over 24 hours. Patients doors were open in the initial measurements. Next, three 1-hour education sessions were conducted by a surgeon and nursing supervisor to review noise-reduction strategies with the staff. These included using pagers in vibrate mode, minimizing overhead announcements, and conducting nurse reports and physician teaching sessions in classrooms away from the nurses' station. Finally, the doors were closed except as visitors and staff entered the room. Little impact was seen from staff education. Closing patient doors on surgical floors decreased noise an average of 6.0 dB, a change that patients can readily perceive. Conversely, intensive care unit patients are exposed to more noise with closed doors, presumably because most noise emanates from equipment within the room. A policy of closing patient floor room doors may increase patient satisfaction. PMID- 9731823 TI - Evaluation of acute mental status change in the nonhead injured trauma patient. AB - Acute mental status change in the first 24 hours after trauma is uncommon in nonhead injured patients who initially present with a normal sensorium. Although arterial hypoxemia is the classic etiology for such a mental status change, three less common etiologies should always be considered: cerebral fat embolism, blunt carotid artery injury, and vertebrobasilar artery thrombosis. Prompt diagnosis and appropriate treatment can significantly improve patient morbidity and mortality. Three nonhead injured trauma patients are described illustrating cerebral fat embolism, blunt carotid artery injury, and vertebrobasilar artery thrombosis as causes of acute mental status change. Each patient initially presented with a clear sensorium, but subsequently developed neurological deficits within 24 hours after admission. All had a normal admission CT scan of the head. MRI or conventional arteriography was diagnostic in each case. Any patient who is initially lucid and subsequently develops a neurological deficit, or a patient whose neurological status does not correlate with brain CT findings should undergo immediate evaluation for possible cerebral fat embolism or cervical vessel injury. An algorithm for management of nonhead injured trauma patients with acute mental status deterioration is presented. PMID- 9731824 TI - Positron emission tomography in diagnosis of occult adenocarcinoma of the breast. AB - Occult adenocarcinoma with clinically apparent axillary lymphadenopathy represents a challenging surgical problem. Mammography is frequently unable to identify a primary breast carcinoma, and extramammary sources are common and equally difficult to identify. This may leave the clinician and patient with a conundrum of whether to proceed with "blind" mastectomy. A 35-year-old white female presented with axillary adenopathy and a normal breast physical exam. Mammography was unable to demonstrate a specific tumor. Excisional biopsy of the axillary lymph node demonstrated metastatic adenocarcinoma. Positron emission tomography showed increased uptake in the breast and the axilla, consistent with breast carcinoma and axillary metastases. The patient underwent modified radical mastectomy and pathologic review of the specimen proved infiltrating ductal carcinoma in the breast with metastatic nodes. Positron emission tomography may be helpful in localizing occult carcinoma of the breast that presents with metastatic lymph nodes and in excluding other potential primaries. PMID- 9731825 TI - Zenker's diverticulum in the elderly: a neurologic etiology? AB - Though described in 1769, the etiology of Zenker's diverticulum remains unclear. Various primary esophageal motor disorders have been proposed, but no consistent manometric pattern or anatomic etiology has been uniformly recognized. An association with clinical neurologic disease at our institution prompted a review of 12 cases of Zenker's diverticulum in patients over 60 years of age, treated in the last 8 years. Nine patients (75%) underwent cricopharyngeus myotomy and diverticulectomy, with uniformly good results. Ten patients (83%) had an associated neurologic disorder, substantiated by cranial CT or MRI, in most cases. A wide range of neurologic problems were identified, but a strong trend toward brainstem or basilar lesions was present. As expected, the etiology of the neurologic abnormality in most patients in this group was cerebrovascular disease, but two patients had peripheral neuropathies. We suggest that the etiology of Zenker's diverticulum in the elderly may be neurologic in origin. Esophageal motor disorders, including incomplete upper esophageal sphincter opening and increased hypopharyngeal pressures, which may result in Zenker's diverticulum, may be a manifestation of central or peripheral neurologic disease in the elderly. PMID- 9731826 TI - Re: Videothorascopic removal of a mediastinal teratoma. PMID- 9731827 TI - Clinical pathways for general surgeons: elective ventral (incisional) hernia repair. PMID- 9731828 TI - Transactions of the Sixtieth Annual Meeting of The South Atlantic Association of Obstetricians and Gynecologists. What goes around comes around. Presidential address. PMID- 9731829 TI - A prospective study of transvaginal hydrosonography in the evaluation of abnormal uterine bleeding. AB - OBJECTIVE: The purpose of this study was to determine whether the intrauterine instillation of saline solution during transvaginal ultrasonographic imaging (hydrosonography) improves the diagnostic accuracy in detecting intrauterine abnormalities determined by direct visualization of the intrauterine cavity with either hysteroscopy or after hysterectomy. STUDY DESIGN: This study was a prospective, blinded study of 39 women referred with the diagnosis of abnormal uterine bleeding after failing medical management. A routine vaginal probe ultrasonographic examination was followed by a blinded transvaginal hydrosonography in patients proceeding to either hysteroscopy or hysterectomy. RESULTS: Twelve of the 39 patients had masses that impinged on the intrauterine cavity at hysteroscopy or hysterectomy. In 4 of the 12 patients with confirmed masses, an intrauterine lesion was detected by hydrosonography that was not seen on routine vaginal probe ultrasonography. In no case was an intrauterine mass detected by the hysteroscopy or after hysterectomy when hydrosonography indicated a normal intrauterine cavity. In 4 cases hydrosonography suggested that masses were present that were not confirmed at direct visualization. Although hydrosonography always recognized when intracavitary pathologic conditions existed in a patient, hydrosonography sometimes underestimated the number of intracavitary lesions present. CONCLUSIONS: Hydrosonography is a simple, minimally invasive, and effective tool to use in the evaluation of patients with abnormal uterine bleeding who have not responded to medical treatment. In no case did hydrosonography fail to indicate when pathologic conditions existed in a given patient, and a normal hydrosonography always indicated a normal intrauterine cavity at hysteroscopy or after hysterectomy. For these reasons hydrosonography is a sensitive tool to triage women with abnormal uterine bleeding to operative or conservative management. PMID- 9731830 TI - Outcome comparison of in vitro fertilization treatment with highly purified subcutaneous follicle-stimulating hormone (Fertinex, a urofollitropin) versus intramuscular menotropins. AB - OBJECTIVE: The aim of this study was to investigate various outcome measures of stimulation with highly purified subcutaneous follicle-stimulating hormone (Fertinex, a urofollitropin) compared with first- and second-generation urinary human menopausal gonadotropin standards (Pergonal, Metrodin). STUDY DESIGN: Retrospective analysis was restricted to our most efficient in vitro fertilization age group (23-34 years). Data from Institute for Assisted Reproduction in vitro fertilization cycles 1 through 11 with Pergonal, Metrodin, or both were tabulated for hormonal values, oocyte quality, and embryo outcome as baseline data. Patients in cycles 12 through 13 were treated with Fertinex and Pergonal or Fertinex alone and then reviewed for the same parameters. RESULTS: Two hundred thirty-eight in vitro fertilization records with embryo transfer were analyzed. Clinical pregnancy rates per embryo transfer in an optimal age group were similar despite use of first- through third-generation urinary gonadotropin preparations: Pergonal and Metrodin, 67%; Metrodin, 64%; Fertinex and Pergonal, 62%; and Fertinex, 54%. There were no discernible differences in hormonal response, oocyte recovery, or embryonic growth. CONCLUSION: Administered subcutaneously, the third-generation urinary gonadotropin preparation Fertinex is effective in in vitro fertilization treatment in young women. PMID- 9731831 TI - Ethical issues in managed care for the obstetrician and gynecologist. AB - The practice of medicine is now managed. Of this there is no doubt. The individual physician is placed in an ever-increasingly vulnerable position. He or she must cope with a myriad of contractual arrangements with strange concepts such as "withholds," "capitation," "covered lives," "limited liability on the part of the managed care organization;" "outcomes analysis," "practice guidelines," and, last but not least, "gag rules." Patients are being denied care that the physician may consider, if not essential, at least most desirable. On the one hand, the physician must serve a fiduciary obligation to the patient and act as the patient's advocate; on the other hand, the physician's income may be proportionally dependent on limiting the extent of the patient's access to unlimited care. The physician may be limited by restrictions imposed by the managed care organization as to what disclosures he or she may make to the patient regarding limitations of care. We will explore these issues from an ethical perspective and attempt to offer some insights on the basis of a review of the comments of many knowledgeable commentators on this topic, and we will explore the virtues that physicians will need to rely on to come to grips with the dilemmas they will face in the future with managed care. PMID- 9731832 TI - Minilaparotomy hysterectomy. AB - OBJECTIVE: This article reports our experience with minilaparotomy hysterectomy. STUDY DESIGN: Minilaparotomy was defined as a skin incision < or = 6 cm in length. From January 1, 1996, to June 30, 1997, data were collected on all patients who underwent hysterectomy by means of a minilaparotomy. RESULTS: During the study a total of 250 hysterectomies were performed. Twenty-six of those were performed by means of a minilaparotomy. The mean age of the patients was 54 years. Seven had endometrial cancer and 8 had an adnexal mass. In 1 patient the incision was extended for staging of an ovarian cancer. The only intraoperative complication was rupture of a 6-cm ovarian tumor. After operation, 2 patients had febrile morbidity, 1 had a prolonged ileus, and in 1 atrial fibrillation developed. The mean uterine weight was 123 g. Median day of Foley catheter removal and mean day of ambulation, regular diet, and discharge were 1 day, 1.2 days, 2.4 days, and 3.4 days, respectively. CONCLUSION: Minilaparotomy is a safe and feasible route of hysterectomy for a selected group of patients. PMID- 9731834 TI - Been there--done that: surgical challenges. PMID- 9731833 TI - Expression and specific immunolocalization of the human parathyroid hormone/parathyroid hormone-related protein receptor in the uteroplacental unit. AB - OBJECTIVE: Our purpose in these studies was to determine the expression and cellular localization of the parathyroid hormone/parathyroid hormone-related protein receptor in the human uteroplacental unit. STUDY DESIGN: Human uteroplacental tissues were obtained and ribonucleic acid was extracted. Reverse transcriptase-polymerase chain reaction was performed with use of primers for both the parathyroid hormone/parathyroid hormone-related protein receptor and human phosphoglyceraldehyde dehydrogenase. Ethidium bromide-stained gels and Southern blots were evaluated, and polymerase chain reaction fragments were sequenced. For immunohistochemistry, slides were incubated with a newly developed antibody (3D1.1) specific for the parathyroid hormone/parathyroid hormone-related protein receptor, and bound monoclonal antibody was detected by use of the avidin biotin technique. RESULTS: Reverse transcriptase polymerase chain reaction gels and blots showed that receptor messenger ribonucleic acid was present in choriodecidua, placenta, and myometrium. Sequence analysis revealed complete identity of the receptor product and the known nucleotide sequence in the receptor. There was intense receptor staining of the myometrial smooth muscle as well as staining of the endothelium and smooth muscle of the associated vasculature. In umbilical cord immunoreactive receptor was found in the vascular endothelium and vascular smooth muscle cells and in stromal cells. In choriodecidua receptor was found in chorionic trophoblasts and decidualized endometrial stromal cells. In all tissues immunostaining was specific, as evidenced by the blocking of staining after addition of receptor peptide to the antibody (absorbed controls). CONCLUSION: The parathyroid hormone/parathyroid hormone-related protein receptor is widely expressed in the human uteroplacental unit. The cellular localizations of the receptor in smooth muscle reflect the ability of parathyroid hormone-related protein to relax both uterine and vascular smooth muscle. The presence of novel autocrine and paracrine systems in the human uteroplacental unit is suggested by the finding that the same cells or adjacent cells produce both parathyroid hormone-related protein and its receptor. PMID- 9731835 TI - Follow-up of sexual assault victims. AB - OBJECTIVES: Our goal was to describe medical findings and health-related concerns of sexual assault victims who returned for follow-up and to assess demographic and assault characteristics of victims who used follow-up services compared to those who did not. STUDY DESIGN: This study is a retrospective cohort analysis involving records from two sources: the acute sexual assault evidentiary examination and the SAFE (Sexual Assault Follow-up Evaluation) Clinic visit. Data were extracted from the records of 389 adolescent and adult victims who reported an acute sexual assault and underwent a complete evidentiary examination between January 1, 1995, and June 30, 1997. Descriptive statistics were generated on demographic and historical information, assault characteristics, and medical and laboratory findings. For the subgroup that presented for follow-up, additional descriptive statistics were generated to describe their interim history, concerns, medical outcomes, treatments, and psychosocial functioning. Chi2 analyses were used to identify differences in the group that returned for follow up compared with the group that did not. Statistical significance was defined as P < .05. RESULTS: There were no differences in age, race, or perpetrator factors between patients who used follow-up services and patients who did not return to the SAFE Clinic. Similarly, there were no other assault characteristics, relationships, or physical examination findings that were associated with follow up patterns. A total of 31% (n = 122) of all sexual assault victims returned for a follow-up visit. Physical complaints were reported by 42.6%, but 98.0% had normal findings at a general examination, and 94.8% had a normal result of gynecologic examination. Pregnancy and sexually transmitted diseases, including human immunodeficiency virus, were identified through the follow-up clinic. Since the assault, 49.2% had been sexually active, 10% with multiple partners and 73.3% without consistent condom use. Disturbances in sleep, sexual function, and appetite were commonly reported among victims at follow-up. Numerous assault related fears were reported. CONCLUSIONS: Among recent rape victims, follow-up rates are low, and there are no factors that correlate with the use of follow-up services. Those who do come in for follow-up have physical complaints and health related concerns that are related to their recent assault, but most have normal physical findings. Efforts to reach sexual assault victims will require aggressive and innovative strategies to remain in contact with women and girls after rape. PMID- 9731836 TI - Urinary complications of Miami pouch: trend of conservative management. AB - OBJECTIVE: Continent urinary diversions have become popular among gynecologic oncologists. Much information has been gained concerning the complications and current management of patients with continent ileocolonic reservoirs. The high mortality rate associated with reoperation has led clinicians to adopt a trend toward conservative means of management. The purpose of this study was to evaluate the applicability of conservative management of complications related to the creation of the continent ileocolonic reservoir Miami pouch. STUDY DESIGN: Patients who underwent creation of the Miami pouch at the Division of Gynecologic Oncology, University of Miami School of Medicine, since 1988 have been included in this study. Management of complications, with particular emphasis on the conservative treatment, has been reviewed in detail for each patient. Open surgery and conservative treatment have been compared. RESULTS: Seventy-seven patients underwent creation of the Miami pouch from February 1988 to September 1997. Sixty (77.9%) patients were affected by recurrent cervical cancer; 72 (93.5%) were previously radiated. The perioperative mortality rate was 11.7% (9 patients). Six of these patients died as a result of sepsis; all of them underwent reoperation at least once. The most common urinary complications were ureteral stricture or obstruction (22.1%), difficult catheterization (19.5%), and pyelonephritis (13%). Conservative management strategies used for these complications included percutaneous nephrostomy, stent placement, balloon dilatation, radiologically (ultrasonography, fluoroscopy, computed tomography) guided placement of catheters, and antibiotic treatment. Eighty percent of the complications associated with the ileocolonic reservoir were resolved with conservative treatment, whereas 16.9% required surgical revision. CONCLUSION: On the basis of these findings, conservative management of urinary reservoir complications should always be considered before surgical intervention is attempted. The exact time to engage in open revision should be individualized on the basis of the clinical condition of each patient. It is our belief that the conservative approach should be instituted whenever possible but surgical intervention not be delayed when absolutely indicated. PMID- 9731837 TI - A randomized clinical trial of prostaglandin E2 intracervical gel and a slow release vaginal pessary for preinduction cervical ripening. AB - OBJECTIVE: Our purpose was to compare the efficacy of 2 different prostaglandin E2 delivery methods for preinduction cervical ripening. STUDY DESIGN: Ninety patients admitted for labor induction with a Bishop score <8 were randomized to either 0.5 mg prostaglandin E2 intracervical gel (Prepidil) every 6 hours for 2 doses or 10 mg prostaglandin E2 slow release vaginal pessary (Cervidil). Oxytocin induction was begun after 12 hours. It was estimated that enrollment of 90 women would be required to identify a 30% difference in the percent delivered in <24 hours (1 - beta = .80, alpha = .05). Data were analyzed with use of chi2 analysis or the Student t test. RESULTS: There were 45 subjects in each treatment arm. The percent delivered by 24 hours was 53% with intracervical gel and 63% with vaginal pessary (P = .28). Mean change in Bishop score was 1.8 +/- 1.9 for the intracervical gel versus 3.2 +/- 3.1 for the vaginal pessary (P = .01). No difference was demonstrated in mean time to delivery, 28.3 versus 24.0 hours (P = .19) or percent requiring cesarean section. CONCLUSION: Preinduction cervical ripening with a slow release prostaglandin E2 vaginal pessary resulted in greater change in Bishop score than with intracervical prostaglandin E2. There was a trend toward shorter time to delivery with the pessary. There was no statistically significant difference in percent delivered in <24 hours. PMID- 9731838 TI - Regulation of monocyte to macrophage differentiation by antiglucocorticoids and antioxidants. AB - OBJECTIVE: Our purpose was to examine RU 486 and related compounds on monocyte to macrophage differentiation through scavenger receptors and cellular adhesion. STUDY DESIGN: Human monocytes were isolated, cultured, and treated with dexamethasone, levonorgestrel, RU 486, and other structurally related compounds alone or in combination. Macrophage scavenger receptor activity, inhibited by glucocorticoids and associated in the current literature with macrophage cellular adhesion, was determined in this study by counting the number of adherent cells after treatment. In addition, fluorescent-labeled acetyl-low-density lipoprotein uptake was determined as a function of scavenger receptor biologic activity. RESULTS: Dexamethasone, levonorgestrel (antiglucocorticoid only) and RU 486 (antiglucocorticoid and antioxidant) all significantly decreased adherent macrophages (4%, 52%, and 74% of control). Levonorgestrel, however, demonstrated a marked uptake of fluorescent-labeled scavenger receptor ligand. RU 486 and dexamethasone were antagonistic when combined (P < .001); levonorgestrel was less antagonistic but, however, still significant (P < .05). Reduced RU 486 (antioxidant but loses antiglucocortioid activity) decreased cellular adhesion, yet scavenger receptor function was enhanced. Both probucol (extracellular mechanism of action) and probucol analog (intracellular action) markedly up regulated scavenger function, but once again a separation of adhesion from scavenger activity was noted. Vitamin E (antioxidant) and onapristone (antioxidant and antiglucocorticoid) had virtually little to no effect on adhesion and scavenger receptor activity. Finally, pyrrolidine dithiocarbamate, a potent oxygen-free radical quencher, was toxic to all cells examined. CONCLUSIONS: RU 486 is a known antiglucocorticoid with novel antioxidant properties first demonstrated by our laboratories. Levonorgestrel has antiglucocorticoid but no antioxidant activity. RU 486 antagonized the inhibitory effect of dexamethasone on scavenger receptor development, whereas levonorgestrel was stimulatory. A separation of scavenger receptor-induced cellular adhesion and scavenger receptor internalized ligand was demonstrated by (1) reduced RU 486, which loses its antiglucocorticoid activity but retains its antioxidant activity, and (2) probucol analog, which is chemically altered to allow intracellular entry. Glucocorticoids decrease the development of scavenger receptors, whereas antioxidants regulate inflammatory cytokines by intracellular mechanisms. It is therapeutically important to up-regulate scavenger receptor activity by antiglucocorticoids in the peritoneal cavity of women with endometriosis. However, because these mechanisms also induce inflammatory cytokines, a balance of antioxidants and antiglucocorticoids such as those demonstrated in the above study may prove beneficial. PMID- 9731839 TI - Expression of gamma-glutamyl transpeptidase in stage III and IV ovarian surface epithelial carcinomas does not alter response to primary cisplatin-based chemotherapy. AB - OBJECTIVE: Gamma-glutamyl transpeptidase activity has been shown to be essential for the nephrotoxicity of cisplatin. The purpose of this study was to determine whether expression of gamma-glutamyl transpeptidase in ovarian carcinomas is necessary for the antitumor effect of cisplatin. STUDY DESIGN: Tumor tissue from 18 patients with stage III or IV ovarian serous papillary carcinoma or poorly differentiated adenocarcinoma was analyzed for expression of gamma-glutamyl transpeptidase by histochemical or immunohistochemical staining. Response to cisplatin-based combination chemotherapy was evaluated on the basis of clinical response, progression-free interval, and survival. RESULTS: Gamma-glutamyl transpeptidase expression in the tumors ranged from 0% to 100% of the tumor cells gamma-glutamyl transpeptidase positive. Patient survival ranged from 15 months to 9 years. Twelve of the 18 patients had a complete response to the initial course of cisplatin-based combination chemotherapy. There was no statistically significant correlation between either response or time to relapse and gamma glutamyl transpeptidase expression. However, there was a correlation between high levels of gamma-glutamyl transpeptidase in the tumor and acute ototoxicity in patients treated with cisplatin. Expression of high levels of gamma-glutamyl transpeptidase in the tumor was also found to be associated with shorter patient survival, suggesting that gamma-glutamyl transpeptidase might have a role in resistance to drugs used in second- and third-line therapy. CONCLUSIONS: Expression of gamma-glutamyl transpeptidase in ovarian serous papillary or poorly differentiated adenocarcinomas is not necessary for the antitumor activity of cisplatin. A correlation was found between high levels of gamma-glutamyl transpeptidase in the tumor and both increased ototoxicity from cisplatin and decreased patient survival. These data suggest that administering an inhibitor of gamma-glutamyl transpeptidase activity to block the nephrotoxicity of cisplatin would not interfere with its therapeutic effect. PMID- 9731840 TI - The temporal efficacy of early second-look lysis of adhesions in reducing postoperative adhesions in a murine model. AB - OBJECTIVE: Our aim was to determine whether early second-look lysis of adhesions reduces postoperative adhesions. STUDY DESIGN: With the murine uterine horn model, early second-look lysis of adhesions was performed 5, 7, 14, and 21 days after an electrocautery injury. Sites with adhesions (between 36 and 46/time point) underwent lysis of adhesions. Fourteen days later, a reviewer blinded to the treatment assessed adhesion formation, including adhesions not present at early second-look lysis of adhesions (eg, de novo adhesions). RESULTS: The rate of adhesion formation was 49% of control sites, unchanged when the early second look lysis of adhesions was performed at 5 (44.4%) and 7 (39.5%) days, reduced at 14 days (28.6%), and increased at 21 days (74%). The pattern of de novo adhesions was similar, 17.6% when the early second-look lysis of adhesions was performed at 5 days, 10% at 7 days, 0% at 14 days, and 28.6% at 21 days. The only histologic difference between the groups was neovascularity at day 21. CONCLUSIONS: Early second-look lysis of adhesions was effective in reducing postoperative adhesions only when performed at 14 days in this model, suggesting that the specific cellular events occurring at the time of the early second-look lysis of adhesions are critical to efficacy. PMID- 9731841 TI - Neonatal mortality for very low birth weight deliveries in South Carolina by level of hospital perinatal service. AB - OBJECTIVE: The purpose of this study was to determine whether neonatal mortality rates for very low birth weight (500 to 1499 g) infants born in South Carolina differ by level of perinatal services available at the hospital of birth. STUDY DESIGN: Linked live birth certificates and infant death certificates for 1993 through 1995 were used. Birth weight-specific neonatal mortality rates among 2375 very low birth weight infants were estimated and analyzed by race and by level of perinatal services at the hospital of birth. Rates were compared with chi2 analysis. RESULTS: Seventy-eight percent of very low birth weight deliveries occurred in level III hospitals. The overall neonatal mortality rate was 178 deaths/1000 very low birth weight live births. Neonatal mortality rates, adjusted for birth weight and race, were significantly higher (P < .05) for infants born in level I hospitals (267 deaths/1000 live births), all level II hospitals (232 deaths/1000 live births), and level II hospitals with neonatologists (213 deaths/1000 live births) than for infants born in level III centers (146 deaths/1000 live births). CONCLUSION: Very low birth weight infants are more likely to survive if born in level III hospitals than in level I or II facilities, with or without neonatologists. Obstetric providers should support public health efforts and perinatal health systems to ensure that all women have access to a strong system of risk-appropriate perinatal care. PMID- 9731842 TI - Atypical glandular cells of undetermined significance: a five-year retrospective histopathologic study. AB - OBJECTIVE: Our purpose was to ascertain the types and frequency of pathologic conditions associated with atypical glandular cells of undetermined significance on Papanicolaou smears. STUDY DESIGN: A 5-year retrospective review of screening cervical cytologic examinations diagnosed as atypical glandular cells of undetermined significance was performed at the University of Virginia to determine pathologic findings associated with atypical glandular cells of undetermined significance on Papanicolaou smears stratified by subtype and overall. RESULTS: Pathologic findings for the respective Papanicolaou smears with the diagnosis of atypical glandular cells of undetermined significance not otherwise specified, favor benign, squamous intraepithelial lesions, and favor neoplasia through the follow-up interval were as follows: squamous intraepithelial lesions in 11%, 8%, 38%, and 20%; adenocarcinoma in situ in 3%, 0%, 0%, and 10%; endometrial hyperplasia in 3%, 5%, 1%, and 2%; and cancer in 8%, 3%, 1%, and 7%. Overall, 63 patients (32%) had preinvasive or invasive lesions. CONCLUSIONS: Atypical glandular cells of undetermined significance on Papanicolaou smears were correlated with significant findings in 45% of patients (32% with preinvasive or invasive lesions and 13% with benign lesions). A prompt and aggressive workup is recommended. PMID- 9731843 TI - Death of the Papanicolaou smear? A tale of three reasons. AB - The Papanicolaou smear is arguably the most cost-effective cancer screening test ever devised. Yet future availability of this low-cost test is seriously threatened by increasing litigation, huge awards, and the implied linkage between "error" and "negligence." The expectation of a 0 error standard, even for a screening test, is central in the current medical-legal climate. Three reasons for this escalating problem will be explored, as follows: (1) substandard laboratories; (2) misunderstanding of the Papanicolaou smear by the media, the public, the legal profession, and physicians, compounded by the "greed factor"; (3) an "acceptable error rate" in quality laboratories for Papanicolaou smear screening. I will explore the future of the Papanicolaou smear and will make specific recommendations for all obstetricians-gynecologists. PMID- 9731844 TI - Understanding miscarriage or insensitive abortion: time for more defined terminology? AB - A search of the two major journals of obstetrics and gynecology published in the United States has shown that the word abortion rather than miscarriage to describe early pregnancy loss is 6 times more likely to be used compared with most other English language journals over the last 5 years. The use of miscarriage, with descriptive adjectives such as threatened, incomplete, and complete, is recommended and the term delayed miscarriage is suggested in place of missed abortion. PMID- 9731845 TI - Can the number of cigarettes smoked predict high-grade cervical intraepithelial neoplasia among women with mildly abnormal cervical smears? AB - OBJECTIVE: We investigated whether the number of cigarettes smoked by women who had mildly abnormal cytologic study results could identify those at risk for high grade dysplasia. STUDY DESIGN: This was a prospective study of all women who were referred for colposcopy with a mildly abnormal cervical smear over a 4-year period. A detailed questionnaire was completed that evaluated sociodemographic characteristics including smoking history. Colposcopy then determined the degree of disease. RESULTS: One hundred seventy-three women were referred with a mildly abnormal cervical smear. There was a significant relationship between the numbers of cigarettes smoked and the risk of high grade disease (P = .007). Once the number of cigarettes smoked daily exceeded 20, the risk of high grade cervical intraepithelial neoplasia was increased fivefold (odds ratio 5.85 [95% confidence interval 1.92 to 17.80]). CONCLUSION: Cigarette smoking is associated with a dose dependent increased risk of cervical intraepithelial neoplasia grade 2 or 3 among women who have mildly abnormal cervical smears. PMID- 9731846 TI - Risk factors for familial and sporadic ovarian cancer among French Canadians: a case-control study. AB - OBJECTIVE: The objective was to compare risk factors between familial and sporadic ovarian cancer by means of a case-control approach. STUDY DESIGN: We conducted a case-control study among French Canadian women in Montreal during 1995-1996. One hundred seventy women 20 to 84 years old with histologically confirmed diagnoses of primary ovarian carcinomas or borderline tumors were interviewed concerning their reproductive, family, and medical histories. During the same period 170 randomly selected population control subjects, frequency matched to the case patients according to age and ethnic group, were also interviewed. Unconditional logistic regression methods were used for data analysis. RESULTS: The major factors influencing the risk of development of ovarian cancer were as follows: (1) family history of breast or ovarian cancer, (2) a late age at use of oral contraceptives (a protective effect), and (3) a late age at last childbirth (a protective effect for familial case patients only). CONCLUSION: These factors had equally great impacts in familial and sporadic cases, implying that the underlying mechanisms of carcinogenesis in sporadic and familial ovarian cancer may be similar and that hereditary ovarian cancer may be preventable. PMID- 9731847 TI - The cost-effectiveness of treating women with a cervical vaginal smear diagnosis of atypical squamous cells of undetermined significance. AB - OBJECTIVE: The purpose was to determine the optimal treatment protocol for women with a cervical vaginal diagnosis of atypical squamous cells of undetermined significance. STUDY DESIGN: By means of decision analysis, 8 strategies were compared in terms of cost, life expectancy, cost-effectiveness, and the number of cancers and complications from treatment. Data were obtained from the medical literature and the University of Iowa Hospitals and Clinics. RESULTS: Compared with more aggressive strategies, such as those that use immediate colposcopy, strategies featuring repeated smears were less expensive and carried fewer complications but had lower life expectancies per patient and more cancers. The strategy of repeating a smear annually had a lower cost per patient than did the other strategies, ranging from $112 to $989, and had a similar discounted life expectancy to that of the strategy with the longest discounted life expectancy. CONCLUSIONS: In most clinical scenarios strategies that used repeated smears were the most cost-effective. PMID- 9731848 TI - Qualification of atypical squamous cells of undetermined significance in an independent laboratory: is it useful or significant? AB - OBJECTIVES: Correlation of cervical smears and biopsy samples emphasizes the significance of atypical squamous cells of undetermined significance as a diagnostic category. STUDY DESIGN: A retrospective study (October 1, 1995-March 21, 1997) was performed on all Papanicolaou smears diagnosed at DIANON Systems, Inc (Stratford, Conn). RESULTS: During this period 1551 patients with Papanicolaou smears had subsequent cervical biopsies performed within 90 days of cytologic sampling. There were 560 diagnoses of atypical squamous cells of undetermined significance. Tissue specimen results were positive in 203 cases (36.2%) and negative in 357 cases (63.8%). Of the positive biopsy results after a smear with atypical squamous cells of undetermined significance, 109 showed low grade squamous intraepithelial lesions and 94 showed high-grade squamous intraepithelial lesions. CONCLUSIONS: This study, the largest cytologic and histologic correlation of atypical squamous cells of undetermined significance, demonstrates that a significant percentage of smears with atypical squamous cells of undetermined significance represent squamous intraepithelial lesions and that qualification of atypical squamous cells of undetermined significance as reactive or dysplastic is useful and should serve as a guide for patient management. PMID- 9731849 TI - Elevated high-density lipoprotein cholesterol and dietary fat intake in women with cyclic mastopathy. AB - OBJECTIVE: This study was designed to examine the contribution of plasma lipids to the pathophysiology of cyclic mastopathy, before and after consideration of diet and sex hormones. STUDY DESIGN: Thirty-four women with severe cyclic mastopathy (case patients) and 29 women without cyclic mastopathy (control subjects) recorded their breast symptoms daily during 1 menstrual cycle. During each menstrual phase (follicular, early luteal, late luteal, and menstrual) they prospectively completed 2 24-hour dietary diaries, provided blood for lipid and hormone assays, and underwent anthropometric measurements. RESULTS: Mean age was 34 years. Premenstrual breast swelling and tenderness were significantly more severe in case patients (P < .0001). Cyclic change (late luteal vs follicular) of high-density lipoprotein cholesterol differed between case patients and control subjects, with case patients having a relative excess of high-density lipoprotein cholesterol in the premenstrual phase (P = .01). Dietary fat intake was greater throughout the cycle in case patients (37.5 vs 33.7% of calories, P = .02), and case patients reported increased appetite in the premenstrual phase (P = .01). In multivariate analyses the contributions of mean dietary fat intake and of cyclic change in high-density lipoprotein cholesterol were independently significant, with odds ratios for upper versus lower quintiles being slightly >5. CONCLUSIONS: Women with cyclic mastopathy had a relative excess of high-density lipoprotein cholesterol during the symptomatic late luteal phase of the menstrual cycle and a higher fat intake throughout the cycle than did control subjects. These observations support the hypothesis that lipids (notably high-density lipoprotein cholesterol) and a high-fat diet play a role in the pathophysiologic characteristics of cyclic mastopathy. PMID- 9731850 TI - The effect of monocyte chemotactic protein 1 in intraperitoneal adhesion formation in a mouse model. AB - OBJECTIVE: Intraperitoneal adhesions are a significant cause of morbidity among women of reproductive age. Monocyte chemotactic protein 1 plays a role in the chemotaxis of mononuclear cells and fibroblasts into the peritoneal injury site. To evaluate its role in intraperitoneal adhesion formation, we used an established postsurgical adhesion model in mice. STUDY DESIGN: Surgical adhesions in mice were induced by scraping and crushing peritoneal sites. We analyzed the injury sites for the temporal expression of monocyte chemotactic protein 1 messenger ribonucleic acid and cellular infiltration at 12 to 24 hours across 10 days and evaluated the effects of monocyte chemotactic protein 1 and anti monocyte chemotactic protein 1 neutralizing antibody on adhesion formation. On induction of adhesions animals were randomly assigned to 1 of 4 groups: (1) control, (2) nonspecific immunoglobulin G, (3) monocyte chemotactic protein 1, (4) anti-monocyte chemotactic protein 1 antibody. Animals received daily intraperitoneal injections for 6 days. Adhesions were scored on day 14 and immunostained for fibroblasts and macrophages. RESULTS: Adhesions developed consistently by the fifth postoperative day. We detected an increase in monocyte chemotactic protein 1 messenger ribonucleic acid expression at 48 hours; this remained until the fourth postoperative day. By the second day macrophages were present at the injury site. Animals treated with anti-monocyte chemotactic protein 1 antibody had significantly fewer adhesions develop than did the other three groups. CONCLUSION: This study demonstrates that monocyte chemotactic protein 1 may play a role in adhesion formation. Inhibiting the action of this chemokine may help to prevent adhesions. PMID- 9731851 TI - Calcium carbonate and the premenstrual syndrome: effects on premenstrual and menstrual symptoms. Premenstrual Syndrome Study Group. AB - OBJECTIVE: Previous reports have suggested that disturbances in calcium regulation may underlie the pathophysiologic characteristics of premenstrual syndrome and that calcium supplementation may be an effective therapeutic approach. To evaluate the effect of calcium carbonate on the luteal and menstrual phases of the menstrual cycle in premenstrual syndrome, a prospective, randomized, double-blind, placebo-controlled, parallel-group, multicenter clinical trial was conducted. STUDY DESIGN: Healthy, premenopausal women between the ages of 18 and 45 years were recruited nationally across the United States at 12 outpatient centers and screened for moderate-to-severe, cyclically recurring premenstrual symptoms. Symptoms were prospectively documented over 2 menstrual cycles with a daily rating scale that had 17 core symptoms and 4 symptom factors (negative affect, water retention, food cravings, and pain). Participants were randomly assigned to receive 1200 mg of elemental calcium per day in the form of calcium carbonate or placebo for 3 menstrual cycles. Routine chemistry, complete blood cell count, and urinalysis were obtained on all participants. Daily documentation of symptoms, adverse effects, and compliance with medications were monitored. The primary outcome measure was the 17-parameter symptom complex score. RESULTS: Seven hundred twenty women were screened for this trial; 497 women were enrolled; 466 were valid for the efficacy analysis. There was no difference in age, weight, height, use of oral contraceptives, or menstrual cycle length between treatment groups. There were no differences between groups in the mean screening symptom complex score of the luteal (P = .659), menstrual (P = .818), or intermenstrual phase (P = .726) of the menstrual cycle. During the luteal phase of the treatment cycle, a significantly lower mean symptom complex score was observed in the calcium-treated group for both the second (P = .007) and third (P < .001) treatment cycles. By the third treatment cycle calcium effectively resulted in an overall 48% reduction in total symptom scores from baseline compared with a 30% reduction in placebo. All 4 symptom factors were significantly reduced by the third treatment cycle. CONCLUSIONS: Calcium supplementation is a simple and effective treatment in premenstrual syndrome, resulting in a major reduction in overall luteal phase symptoms. PMID- 9731852 TI - Antepartum cervical ripening: applying prostaglandin E2 gel in conjunction with scheduled nonstress tests in postdate pregnancies. AB - OBJECTIVE: Our purpose was to evaluate whether inserting prostaglandin E2 gel at the time of scheduled nonstress tests in patients with postdate pregnancies can decrease rates of intervention. STUDY DESIGN: A multicenter pilot study enrolled women with postdate pregnancies with Bishop score < or = 6 who were undergoing antepartum fetal heart rate testing. Patients were randomized in a double-blind fashion to receive either a prostaglandin E2 intracervical gel (Prepidil) or a placebo gel after each of their scheduled nonstress tests. RESULTS: There were no significant differences in the number of antepartum tests, labor inductions, or cesarean sections, the maximum oxytocin dosage, or the interval from admission to delivery in the prostaglandin E2 gel and placebo gel groups (n = 90). In the subset of patients with a Bishop score between 3 and 6 (63 patients), there were fewer inductions in the prostaglandin E2 group (30% vs 55%, P < .05). CONCLUSION: Application of prostaglandin E2 gel at the time of scheduled antepartum testing in patients with postdate pregnancies with unfavorable cervices decreased the induction rate only among patients with intermediate Bishop scores. PMID- 9731853 TI - Methadone maintenance in pregnancy: a reappraisal. AB - OBJECTIVE: The purpose of this study is to evaluate women receiving methadone maintenance during pregnancy. STUDY DESIGN: Thirty-two pregnancies in women receiving methadone maintenance were matched by gestational age to women with a positive urine screen for cocaine at delivery and to drug-free controls. Pregnancy outcome variables were compared, including birth weight and neonatal morbidity. Analysis was by chi2 and t test with significance set at .05. RESULTS: Birth weight of methadone-exposed infants was 2748 g versus 2925 g for cocaine and 3032 g for controls, P = not significant. Birth weight comparison with a 50 mg maternal methadone cutoff dose was not different. A head circumference for methadone infants of 32.4 +/- 4.7 cm was significantly less than controls, 33.5 +/- 4.0 cm, P < .04, but not different from infants of cocaine users, 32.8 +/- 3.1 cm. Women using cocaine had a significantly higher incidence of meconium in labor compared with methadone and controls. Of women taking methadone 27 of 32 (84.3%) were positive for other drugs of abuse in the last screen before or at delivery. Cocaine 12 of 32 (37.5%), other opiates 13 of 32 (40.6%), and marijuana 14 of 32 (43.7%) were the most prevalent. Neonatal withdrawal occurred in 23 of 32 (72%) women taking methadone. The neonates of women using < 50 mg of methadone were as likely to withdraw as those women using > or = 50 mg, 61.5% versus 79.0%, P not significant. Three neonates in the methadone group (9.3%) had major congenital anomalies, with 2 of the 3 (66.6%) resulting in mortality. CONCLUSIONS: Birth outcome is not significantly different between methadone and cocaine users. Women receiving methadone maintenance are likely to abuse other illicit drugs. PMID- 9731854 TI - Circulating vascular cell adhesion molecule-1 in pre-eclampsia, gestational hypertension, and normal pregnancy: evidence of selective dysregulation of vascular cell adhesion molecule-1 homeostasis in pre-eclampsia. AB - OBJECTIVE: Our purpose was to investigate circulating levels of vascular cell adhesion molecule-1 in the peripheral and uteroplacental circulations during normotensive and hypertensive pregnancies. STUDY DESIGN: This prospective observational study involved 2 patient groups. Group 1 consisted of 22 women with pre-eclampsia and 30 normotensive women followed up longitudinally through pregnancy and post partum. There were an additional 13 women with established gestational hypertension. Group 2 consisted of 20 women with established pre eclampsia and 19 normotensive control subjects undergoing cesarean delivery. Plasma levels of vascular cell adhesion molecule-1 were measured in blood drawn from the antecubital vein (group 1) and from both the antecubital and uterine veins (group 2). Data were analyzed by analysis of variance. RESULTS: In group 1 vascular cell adhesion molecule-1 levels did not change significantly throughout normal pregnancy and post partum. Women with established pre-eclampsia had increased vascular cell adhesion molecule-1 levels compared with the normotensive pregnancy group (P = .01). Vascular cell adhesion molecule-1 levels were not elevated in women with established gestational hypertension. In group 2 significantly higher levels of vascular cell adhesion molecule-1 were detected in the uteroplacental (P < .0001) and peripheral (P < .0001) circulations of pre eclamptic women by comparison with normotensive women. In the pre-eclamptic group there was a tendency toward higher vascular cell adhesion molecule-1 levels in the peripheral circulation than in the uteroplacental circulation (P = .06). In contrast to vascular cell adhesion molecule-1, circulating levels of E-selectin and intercellular adhesion molecule-1, other major leukocyte adhesion molecules expressed by the endothelium, were not different in pre-eclamptic and normotensive pregnancies. CONCLUSION: Established pre-eclampsia is characterized by selective dysregulation of vascular cell adhesion molecule-1 homeostasis. This event is not an early preclinical feature of pre-eclampsia, does not persist post partum, is not a feature of nonproteinuric gestational hypertension, and is not observed with other major leukocyte adhesion molecules. Induction of vascular cell adhesion molecule-1 expression in pre-eclampsia may contribute to leukocyte mediated tissue injury in this condition or may reflect perturbation of other, previously unrecognized, functions of this molecule in pregnancy. PMID- 9731855 TI - Ethanol-induced expression of cytokines in a first-trimester trophoblast cell line. AB - OBJECTIVES: Altered cytokine expression at the fetoplacental interface may be a potential mechanism for the development of fetal immune dysfunction in children with fetal alcohol syndrome. This study was conducted to determine whether first trimester trophoblasts respond to ethanol exposure by the induction of specific cytokines. STUDY DESIGN: HTR-8/SVneo trophoblast cells were cultured in vitro in the presence of either ethanol (0.5% [vol/vol]), lipopolysaccharide (1 microg/mL), or ethanol and lipopolysaccharide. Expression of granulocyte colony stimulating factor, regulated on activation normal T cell expressed and secreted, and interleukin-6 was examined by Northern analysis and enzyme-linked immunosorbent assay. RESULTS: Culture in the presence of ethanol, lipopolysaccharide, or lipopolysaccharide and ethanol resulted in the increased transcription and secretion of granulocyte colony-stimulating factor, regulated on activation normal T cell expressed and secreted, and interleukin-6 at significantly greater levels (P < .01) than control cultures. CONCLUSIONS: Human first-trimester trophoblasts express high levels of cytokines when cultured in the presence of ethanol. Trophoblasts may therefore be an important exogenous source of cytokines for the fetus, and altered cytokine levels during early gestation may have an adverse effect on the development of the fetal immune system. PMID- 9731856 TI - Shoulder dystocia and associated risk factors with macrosomic infants born in California. AB - OBJECTIVE: The purpose of this study was to examine the 1-year incidence statewide in California of shoulder dystocia and its associated risk factors. STUDY DESIGN: With a data set that contains computer-linked records from the birth certificate and hospital discharge records of both mother and baby, all births of infants >3500 g in >300 civilian acute care hospitals in California in 1992 were analyzed. All cases of shoulder dystocia were identified from discharge records, birth certificates, or both and were analyzed with both bivariate and multivariate techniques to identify specific risk factors. RESULTS: A total of 175,886 vaginal births of infants >3500 g were included in our database, of which 6238 infants (3%) had shoulder dystocia. The percentages of births complicated by shoulder dystocia for unassisted births not complicated by diabetes were 5.2% for infants 4000 to 4250 g, 9.1% for those 4250 to 4500 g, 14.3% for 4500 to 4750, and 21.1% for those 4750 to 5000 g. Shoulder dystocia increased by approximately 35% to 45% in vacuum- or forceps-assisted births to nondiabetic mothers. Similar increases were seen in unassisted births to diabetic mothers. The risk of shoulder dystocia for assisted births to diabetic mothers was even more dramatic: 12.2% for infants 4000 to 4250 g, 16.7% for those 4250 to 4500 g, 27.3% for those 4500 to 4750 g, and 34.8% for those 4750 to 5000 g. After controlling for other parameters, there was an increased risk of shoulder dystocia associated with diabetes (odds ratio 1.7), assisted delivery (odds ratio 1.9), and induction of labor (odds ratio 1.3). Rates of birth trauma, asphyxia, and length of stay were all increased among births complicated by shoulder dystocia. CONCLUSION: This information on the incidence of shoulder dystocia and associated risk factors for a large statewide population may assist providers of obstetric care in counseling patients when macrosomia is suspected. The inaccuracy of estimating fetal weight is a severe limitation in attempting to establish guidelines designed to prevent shoulder dystocia. PMID- 9731857 TI - In pregnancies with gestational diabetes mellitus and intensive therapy, perinatal outcome is worse in small-for-gestational-age newborns. AB - OBJECTIVE: This study analyzed the relationship between birth weight and perinatal outcome among women with gestational diabetes mellitus. STUDY DESIGN: The relationship between perinatal outcome and birth weight was analyzed for 821 pregnancies of women with gestational diabetes mellitus attended in a tertiary hospital and receiving intensive metabolic therapy (normocaloric diet, self monitoring of blood glucose level and individually tailored insulin regimen when needed). The Mantel-Haenszel test was used to adjust for preterm delivery. RESULTS: Seven percent of neonates were small for gestational age, 85% were appropriate for gestational age, and 8% were large for gestational age. After adjustment for preterm delivery the rates of adverse fetal outcome, low 1-minute Apgar score, and hypoglycemia were greater among small for gestational age neonates than among appropriate and large for gestational age infants (odds ratios 3.08, 2.51, and 3.17, respectively). CONCLUSION: Among women with gestational diabetes mellitus who are receiving intensive therapy, perinatal outcome is worse for small for gestational age neonates than for appropriate and large for gestational age neonates. PMID- 9731858 TI - The acute pressure natriuresis response blunted and the blood pressure response reset in the normal pregnant rat. AB - OBJECTIVES: Our purpose was to test the hypothesis that the acute pressure natriuresis curve was reset in pregnancy to facilitate the volume expansion. STUDY DESIGN: Studies were done with 14- to 16-day pregnant (n = 8) and age matched virgin female (n = 6) Sprague-Dawley rats that were under general anesthesia. The left kidney was denervated, and mechanical clamps were placed on the aorta above and below the renal arteries for manipulation of renal perfusion pressure. Rats received intravenous 0.9% sodium chloride (1.5% body weight/h) and a cocktail of vasoactive factors to suppress variation in endogenous hormones. Renal perfusion pressure was varied acutely from 125 to 95 mm Hg, and glomerular filtration rate, renal plasma flow, sodium excretion, and urine flow were measured in both kidneys at each renal perfusion pressure. Data were analyzed by unpaired t test and by homogeneity by slopes. RESULTS: The acute pressure natriuresis curve was blunted in pregnant rats versus virgins, and the renal nerves were not responsible. The blunted natriuretic response in pregnancy was due to loss of tubular epithelial responsiveness to increased blood pressure. CONCLUSION: The pressure natriuretic response is markedly blunted in pregnancy, permitting the cumulative plasma volume expansion to occur. Contrary to nongravid states, blunting of the acute pressure natriuresis curve in pregnancy is not associated with increased blood pressure because of the profound peripheral vasodilation. This suggests an alteration in the mechanism(s) normally linking blood pressure control to the acute pressure natriuresis relationship. PMID- 9731859 TI - Parathyroid hormone-related peptide (1 to 34) inhibits in vitro oxytocin stimulated activity of pregnant baboon myometrium. AB - OBJECTIVES: We sought to determine the in vitro effect of parathyroid hormone related protein fragment 1 to 34 on oxytocin precontracted myometrium from baboons in late gestation and to explore possible regional uterine differences in responsiveness comparing muscle strips from the lower uterine segment, posterior and anterior corpus, and fundus. STUDY DESIGN: We used cumulative concentration response (1 to 100 nmol/L) curves to parathyroid hormone-related protein (1 to 34) in isolated strips of baboon myometrium obtained at cesarean hysterectomy of 6 pregnant baboons in the last third of pregnancy. RESULTS: Parathyroid hormone related protein (100 nmol/L) decreased both amplitude and frequency of contraction. The maximum effect of parathyroid hormone-related protein on the amplitude of contractions was greater than saline solution (2-way analysis of variance, F ratio 424.0, P < .001), but there was no difference comparing the four regions of the uterus (F ratio 1.342, P = .286). The maximum effect of parathyroid hormone-related protein on the frequency of contractions was greater than saline solution (2-way analysis of variance, F ratio 162.5, P < .0001), but no difference in response was noted in the 4 regions of the uterus (F ratio = 0.682, P = .572). CONCLUSION: Parathyroid hormone-related protein completely inhibited the contractile effect of high doses of oxytocin in the lower segment, corpus, and fundus of the baboon uterus. No difference in response of myometrium was obtained from different regions of the uterus. PMID- 9731860 TI - Uterine relaxation responses to calcitonin gene-related peptide and calcitonin gene-related peptide receptors decreased during labor in rats. AB - OBJECTIVES: Our purpose was to investigate (1) whether uterine relaxation responses to calcitonin gene-related peptide are differentially regulated during pregnancy and labor, (2) the involvement of nitric oxide in smooth muscle relaxant action of calcitonin gene-related peptide in the rat uterus, (3) whether receptors for calcitonin gene-related peptide are expressed in rat uterus, and if so (4) whether the concentrations of these receptors are differently regulated during pregnancy and labor. STUDY DESIGN: Rats were killed on day 18 of gestation, at the time of spontaneous labor, or postpartum day 2. The uteri were removed for in vitro contractility measurements, nitric oxide production, and calcitonin gene-related peptide receptor binding assay. RESULTS: (1) Calcitonin gene-related peptide induced a dose-dependent relaxation in spontaneously contracting uterine strips from pregnant rats on day 18 of gestation; (2) the relaxation effects of calcitonin gene-related peptide on the uterus were decreased during spontaneous delivery at term and post partum compared with that during pregnancy; (3) calcitonin gene-related peptide-induced relaxation was inhibited by pretreatment of the uterine tissue with a calcitonin gene-related peptide receptor antagonist, calcitonin gene-related peptide(8-37); (4) nitric oxide synthesis inhibitor (N(G)-nitro-L-arginine methyl ester) and soluble guanylate cyclase inhibitor (LY83583) significantly decreased calcitonin gene related peptide-induced relaxation of the rat uterus during pregnancy; (5) calcitonin gene-related peptide increased the uterine nitric oxide production in pregnant rats, and this increase was obliterated in the presence of calcitonin gene-related peptide(8-37); and (6) calcitonin gene-related peptide receptors are present in rat uterus, and the concentration of these receptors dramatically increases during pregnancy and decreases during labor at term. CONCLUSIONS: Calcitonin gene-related peptide inhibits uterine spontaneous contractions in rats during pregnancy but not during labor and post partum. The inhibitory effects of calcitonin gene-related peptide on uterine contractility appear to be modulated, at least in part, by the activation of nitric oxide generation in the rat uterus. Changes in calcitonin gene-related peptide receptors could contribute to the changes in calcitonin gene-related peptide-mediated uterine relaxation during pregnancy and labor. PMID- 9731861 TI - Potentially asphyxiating conditions and spastic cerebral palsy in infants of normal birth weight. AB - OBJECTIVE: Our purpose was to examine the association of cerebral palsy with conditions that can interrupt oxygen supply to the fetus as a primary pathogenetic event. STUDY DESIGN: A population-based case-control study was performed in four California counties, 1983 through 1985, comparing birth records of 46 children with disabling spastic cerebral palsy without recognized prenatal brain lesions and 378 randomly selected control children weighing > or = 2500 g at birth and surviving to age 3 years. RESULTS: Eight of 46 children with otherwise unexplained spastic cerebral palsy, all eight with quadriplegic cerebral palsy, and 15 of 378 controls had births complicated by tight nuchal cord (odds ratio for quadriplegia 18, 95% confidence interval 6.2 to 48). Other potentially asphyxiating conditions were uncommon and none was associated with spastic diplegia or hemiplegia. Level of care, oxytocin for augmentation of labor, and surgical delivery did not alter the association of potentially asphyxiating conditions with spastic quadriplegia. Intrapartum indicators of fetal stress, including meconium in amniotic fluid and fetal monitoring abnormalities, were common and did not distinguish children with quadriplegia who had potentially asphyxiating conditions from controls with such conditions. CONCLUSION: Potentially asphyxiating conditions, chiefly tight nuchal cord, were associated with an appreciable proportion of unexplained spastic quadriplegia but not with diplegia or hemiplegia. Intrapartum abnormalities were common both in children with cerebral palsy and controls and did not distinguish between them. PMID- 9731862 TI - An alternative for women initially declining genetic amniocentesis: individual Down syndrome odds on the basis of maternal age and multiple ultrasonographic markers. AB - OBJECTIVE: Our purpose was to develop a method of calculating the individual odds of Down syndrome on the basis of a combination of maternal age and multiple ultrasonographic parameters that can be used to counsel women at high risk who initially decline amniocentesis. STUDY DESIGN: Maternal age and ultrasonographic biometry data were collected prospectively on 3254 normal and 30 Down syndrome singleton fetuses between 15 and 24 weeks' gestation. Humerus length data were expressed as multiples of the normal median. Log transformation of the humerus length data permitted their expression in gaussian frequency distributions and the calculation of likelihood ratios for Down syndrome on the basis of humerus length. We also developed likelihood ratios on the basis of the degree of nuchal skinfold thickening and the presence or absence of hyperechoic fetal bowel and hypoplastic fifth digit. RESULTS: The ultrasonographic parameters and maternal age did not significantly correlate with each other and were significant independent predictors of Down syndrome. We therefore calculated the individual odds of Down syndrome by using the product of the age-related risk and the likelihood ratios associated with nuchal thickening, humerus length shortening, and the presence or absence of hyperechoic fetal bowel or fifth digit hypoplasia, respectively. At a Down syndrome risk level of >1:50, a 60.0% detection rate with 4.5% false-positive rate was observed with a screen-positive rate of 5.5%, positive predictive value of 1:10, and odds ratio (95% confidence interval) of 28.4 (12.8 to 64.0). CONCLUSION: This is the first report of individual odds calculation based on multiple midtrimester biometry parameters and maternal age. The screening efficiency is similar to that reported with triple-analyte serum screening. These data are useful for counseling women who are at increased Down syndrome risk and initially decline amniocentesis. PMID- 9731863 TI - Hemostasis in the uteroplacental and peripheral circulations in normotensive and pre-eclamptic pregnancies. AB - OBJECTIVE: Our purpose was to determine the hemostatic changes in the uteroplacental and peripheral circulations in normotensive and pre-eclamptic pregnancies. STUDY DESIGN: This prospective, observational study involved 2 patient groups. Group 1 consisted of 30 normotensive women and 22 women with pre eclampsia who were followed up longitudinally through pregnancy and post partum. Group 2 consisted of 20 women with established pre-eclampsia and 19 normotensive control subjects, all undergoing cesarean section. Plasma levels of thrombin antithrombin III complex, soluble fibrin, plasmin-alpha2-antiplasmin complex, and fibrin-degradation product (D-dimer) were measured in blood drawn from the antecubital vein (group 1) and from both the antecubital and uterine veins (group 2). Data were analyzed by analysis of variance. RESULTS: In group 1 levels of thrombin-antithrombin III complex, soluble fibrin, and fibrin-degradation product were significantly higher during normal pregnancy than at 6 weeks post partum. Plasmin-alpha2-antiplasmin complex levels did not change. No differences between the pre-eclamptic and normotensive pregnancy groups were found for any of the hemostatic markers. In group 2 normotensive women undergoing cesarean section, thrombin-antithrombin III complex and soluble fibrin levels were significantly higher in the uterine vein than in the antecubital vein. In group 2 women with pre-eclampsia, thrombin-antithrombin III complex and fibrin-degradation product levels were significantly higher in the uterine vein than in the antecubital vein. In addition, plasmin-alpha2-antiplasmin complex and fibrin-degradation product levels were higher and soluble fibrin levels were lower in the uterine vein in the pre-eclamptic group than in the normotensive group. CONCLUSION: Both the coagulation and fibrinolytic systems are activated during normal pregnancy. Activation of these systems is more marked in the uteroplacental circulation than in the systemic circulation in both normotensive and pre-eclamptic pregnancies. An abnormal pattern of hemostasis occurs in the uteroplacental circulation in pre eclampsia. PMID- 9731864 TI - Prevention of herpes simplex virus infection and latency by prophylactic treatment with acyclovir in a weanling mouse model. AB - OBJECTIVE: Acyclovir is an antiviral agent that inhibits acute herpes simplex virus replication and decreases the frequency of reactivation, but it is not currently used to prevent primary disease or the establishment of latency. The purpose of this study was to reexamine the efficacy of acyclovir in preventing acute and latent herpes simplex virus infection. STUDY DESIGN: Mice were infected by footpad inoculation with 2 viral recombinants that express beta-galactosidase. Half of each group was treated prophylactically with intraperitoneal acyclovir and then given acyclovir in the drinking water. Four days after infection, the dorsal root ganglia were removed, fixed, and stained, and the number of cells expressing beta-galactosidase were counted. RESULTS: Compared with placebo, prophylactic acyclovir completely inhibited acute viral replication as evidenced by the absence of beta-galactosidase activity (P < .001) and significantly decreased the number of neurons harboring latent infection (P = .01). CONCLUSION: Acyclovir prophylaxis prevented acute and reduced latent ganglionic infection with herpes simplex virus in a weanling mouse model. PMID- 9731865 TI - Urocortin in pregnancy. AB - OBJECTIVES: The aim of this study was to investigate the possibility that urocortin is the ligand that displaces corticotropin-releasing hormone from its binding protein in the maternal circulation during pregnancy and, if so, to determine whether urocortin, like corticotropin-releasing hormone, is synthesized in substantial quantities in the placenta. STUDY DESIGN: A radioimmunoassay specific for urocortin was developed and used for measurement of the peptide in chorionic villi and fetal membranes (amnion and chorion) from normal and preeclamptic pregnancies. These tissues were also assayed for corticotropin releasing hormone. Assays for urocortin were also carried out on normal term pregnant and nonpregnant myometrium and on plasma from nonpregnant individuals, and assays for both peptides were performed on sequential normal pregnancy plasma samples taken from mid gestation until term. RESULTS: Corticotropin-releasing hormone was present in normal term (1904 +/- 489 pg/g) and preeclamptic placentas (5897 +/- 1526 pg/g) and in normal term fetal membranes (645 +/- 155 pg/g, n = 6 in all cases). Urocortin was not detected in any of the tissues studied, nor was it found in the normal human plasma samples. Unlike the situation for corticotropin-releasing hormone, no pregnancy-related pattern was seen for urocortin in the plasma from pregnant women. CONCLUSIONS: Urocortin is not translated to any great extent in the pregnancy tissues investigated, nor is it present in the circulation of pregnant women in detectable amounts. Furthermore, it is unlikely that urocortin is responsible for the high maternal plasma levels of free corticotropin-releasing hormone circulating in the latter stages of pregnancy, but this does not preclude the possibility that another, as yet uncharacterized, corticotropin-releasing hormone-like peptide may be. PMID- 9731866 TI - Doppler flow velocity waveforms of the umbilical arteries correlate with intravillous blood volume. AB - OBJECTIVE: My purpose was to measure the volume of the fetoplacental vessel tree and to relate findings to Doppler flow patterns of the umbilical arteries. STUDY DESIGN: One hundred sixty placentas were examined by means of standardized random block placental histomorphometry after delivery and the results were compared with antenatal Doppler findings. RESULTS: There was a high correlation (r = 0.703) between the intravillous blood volume obtained from measurements of intermediate and terminal villi and the Doppler flow velocity waveforms detected within the last week before delivery. Moreover, the reduced size of a vessel tree less than 85 mL is highly predictive of perinatal complications, such as fetal growth restriction, low umbilical artery pH values after birth, reduced Apgar scores, and cesarean section for fetal distress. CONCLUSION: These data suggest that reduced end-diastolic flow velocities in the umbilical arteries are associated with elevated fetoplacental impedance owing to reduced vascularization of intermediate and terminal villi. PMID- 9731867 TI - Cervical screening adjuncts: recent advances. AB - In an effort to reduce the false-negative rate of cervical cytologic findings, several new technologies have recently evolved. Automated cytologic testing (PapNet, AutoPap 300 QC) proposes to rescreen negative conventional cytologic findings to identify smears likely to be false negative. Fluid-based monolayers (ThinPrep, CytoRich) propose to reduce the false-negative rates by optimizing the collection and preparation of cells. Human papillomavirus deoxyribonucleic acid testing by Hybrid Capture has been proposed for a variety of screening and triage roles. Visual screening after application of acetic acid is done by cervicography by use of a photographic technique, whereas in speculoscopy the screening is done by direct visualization of the cervix by the primary care provider. Polarprobe uses biophysical parameters and a computer algorithm to give an instantaneous prediction of the likelihood of cervical disease. Each of these techniques, as well as the clinical experience with them, is reviewed. Current and possible future uses are discussed with regard to both clinical usefulness and cost effectiveness. PMID- 9731868 TI - Recurrent candidiasis and "malcarbohydrate metabolism". PMID- 9731869 TI - Fetal body composition and ultrasonographic estimates of fetal weight. PMID- 9731870 TI - Risk of magnesium for respiratory failure in pregnancy. PMID- 9731871 TI - Nonobstetric emergencies in pregnancy. PMID- 9731872 TI - Databases and the statistical usage of (perinatal) results--reply. PMID- 9731873 TI - Some concerns about the new research guidelines for interpretation of electronic fetal heart rate monitoring. PMID- 9731874 TI - Erb's palsy without shoulder dystocia. PMID- 9731875 TI - Clinical outcome of mild fetal ventriculomegaly. PMID- 9731876 TI - Are there seasonal effects on obstetric circadian rhythms? PMID- 9731877 TI - Maternal weight, body mass index, and cesarean delivery. PMID- 9731878 TI - Cesarean section. PMID- 9731879 TI - Papillary adenoma of the large intestine: a historical perspective. PMID- 9731880 TI - Prevention of osteoporosis in women treated for hereditary breast and ovarian carcinoma: a need that is overlooked. PMID- 9731881 TI - Predicting outcomes of therapy for prostate carcinoma patients. PMID- 9731882 TI - Cost-effectiveness of hepatic artery infusion chemotherapy. PMID- 9731883 TI - Interphase cytogenetics for studying solid tumors. PMID- 9731884 TI - Staging in thyroid carcinoma. PMID- 9731885 TI - Staging of thyroid carcinoma--reply. PMID- 9731886 TI - Expression of P-glycoprotein, a multidrug-resistance gene product, is induced by radiotherapy in patients with oral squamous cell carcinoma. AB - BACKGROUND: It has been observed that patients who have previously undergone radiotherapy have a lower rate of response to chemotherapy. METHODS: The authors investigated the effects of radiation on the expression of P-glycoprotein (Pgp), a multidrug-resistance gene product, in 56 patients with primary oral cancer. No patients received prior or concurrent chemotherapy. The 56 patients consisted of 3 groups: 1) 20 patients with preradiation or pretreatment specimens only, 2) 18 patients with both pre- and postradiation specimens, and 3) 18 patients with postradiation specimens only. Pgp expression was determined by immunohistochemistry with two monoclonal antibodies, C219 and C494. RESULTS: Among patients in Groups 1 and 2, only 1 (2.6%) and 2 (5.3%) patients had Pgp expression in their tumors before treatment with C219 and C494, respectively. For Group 2 patients, 66.7% and 72.2% had tumors that expressed Pgp with the two antibodies, respectively, only after and not prior to radiation. When patients in Groups 2 and 3 were combined, 63.9% and 72.2% had Pgp expression with the two antibodies, respectively, after radiation. Pgp expression was significantly induced after radiation compared with expression before treatment (P < 0.001). Overexpression of p53 protein, detected by immunohistochemistry with DO-7 antibody, was seen in the tumors of 40 patients (71.4%), and the status was quite consistent throughout radiotherapy. Pgp expression had no significant association with p53 protein expression. CONCLUSIONS: Pgp expression was significantly induced by radiation in human oral cancers. This induction of Pgp expression likely confers multidrug resistance to the cancer cells and may affect the efficacy of subsequent or concurrent chemotherapy. It may explain the lower rate of response to chemotherapy among patients who have previously had radiotherapy. PMID- 9731887 TI - Micrometastasis and tumor cell microinvolvement of lymph nodes from esophageal squamous cell carcinoma: frequency, associated tumor characteristics, and impact on prognosis. AB - BACKGROUND: The purpose of this study was to investigate micrometastasis (MM) and tumor cell microinvolvement (TCM) in the regional lymph nodes of patients with esophageal squamous cell carcinoma (SCC). METHODS: MM was defined as individual tumor cells or tumor cell clusters <0.5 mm in greatest dimension with a surrounding stromal reaction. TCM was defined as individual tumor cells or tumor cell clusters without a surrounding stromal reaction. One thousand nine hundred and fifty-four lymph nodes were dissected from 69 complete (R0) resection specimens of TNM classified pT1-3, pN0 or pN1, and M0 esophageal SCC. These lymph nodes were examined immunohistochemically using the monoclonal antibody cocktail AE1/AE3 for cytokeratins. The primary tumors were immunostained with an anti-E cadherin monoclonal antibody. RESULTS: MM +/- TCM was found in 13 cases (31.7%) and TCM alone in 2 cases (4.9%) of the 41 pN0 cases. The pN0 patients with MM (but not TCM) had the same shorter survival as the original pN1 cases (P < 0.05). Of the 69 primary tumors, 49 (71.0%) had reduced or negative E-cadherin expression that showed a correlation with the occurrence of lymph node metastases (original pN1), MM, and TCM, but not prognosis. CONCLUSIONS: The results of the current study show that, in SCC of the esophagus, MM, but not TCM, in the regional lymph nodes is prognostically equivalent to metastasis and should be examined by immunohistochemistry to classify these cases correctly as pN1. PMID- 9731888 TI - Clonal analysis of superficial depressed-type gastric carcinoma in humans. AB - BACKGROUND: An important unanswered question concerning the histogenesis of superficial-type gastric carcinoma is whether it is monoclonal or multiclonal in origin. Therefore, the authors analyzed multiple areas of each cancer with a clonality assay based on trinucleotide repeat length polymorphism of the human androgen receptor gene (HUMARA) that was subject to random inactivation of X chromosomes. METHODS: The HUMARA assay was applied to 15 gastric carcinomas, early and advanced stage, manifested in superficial, depressed lesions of various sizes and at least some signet ring cells. DNA was extracted from fresh frozen and formalin fixed tumor tissues that were microdissected from the mucosal lesions, and the HUMARA locus was amplified by polymerase chain reaction with and without prior digestion of nonmethylated DNA with Hpa II. The amplified DNA samples were loaded on polyacrylamide gels, electrophoresed, and visualized by a silver-staining method. RESULTS: In the 15 cases examined, 9 cancers were informative (had features of the types sought in this study), and in these 9 cancers a total of 57 areas were analyzed. In 7 of the 9 cancers, the inactivated allele was common to all the informative areas of each tumor, irrespective of the macroscopic shape of the tumor or the degree of histologic heterogeneity within it. In one of the two remaining cancers, the inactivated allele of one of the areas examined was different from those in the other areas. CONCLUSIONS: Most of the superficial depressed-type gastric carcinomas in this study were demonstrated to be of monoclonal origin. This finding supports a notion expressed previously in the literature that superficial-type carcinoma has a long natural history, and it indicates that efforts to detect gastric carcinomas in early stages to improve patients' survival should be encouraged. PMID- 9731889 TI - Increased cell proliferation of the gastric mucosa in first-degree relatives of gastric carcinoma patients. AB - BACKGROUND: Studies not considering Helicobacter pylori infection have suggested the presence of a hereditary risk for gastric carcinoma. However, other studies have identified intrafamilial clustering of H. pylori infection as a causal factor in gastric carcinogenesis. This prompted the authors to study the effect of H. pylori and hereditary factors on the proliferation of gastric mucosa because hyperproliferation appears to be an early step in carcinogenesis. METHODS: In a total of 39 patients (19 first-degree relatives of patients with gastric carcinoma and 20 dyspeptic controls), 2 biopsy specimens each from the antrum and corpus were examined histologically. In addition, crude nuclei fractions were prepared from other biopsy specimens obtained in the same manner. Nuclei were fixed in 70% ethanol and stained with propidium iodine prior to measurement. A cell cycle analysis was performed using a flow cytometer. For analysis a proliferative index (PI) (percentage of nuclei in the S- and G2/M phases) was calculated. RESULTS: In comparison with control patients, first degree relatives of gastric carcinoma patients had increased mucosal proliferation of the antrum (Student's t test, P = 0.017). After excluding patients with H. pylori infection (12 in each group), relatives of gastric carcinoma patients had significantly increased proliferation not only in the antrum (PI: 16.5 vs. 12.1; P = 0.043), but also in the corpus (PI: 17.2 vs. 13.0; P = 0.024). CONCLUSIONS: A family history of gastric carcinoma may increase the risk for developing gastric carcinoma via mucosal hyperproliferation, irrespective of H. pylori infection. PMID- 9731890 TI - Cost-effectiveness of systemic and regional chemotherapy for the treatment of patients with unresectable colorectal liver metastases. AB - BACKGROUND: Management of unresectable colorectal liver metastases (CLM) can be by regional (hepatic arterial infusion [HAI]) or systemic chemotherapy, or by symptom control alone. In this study the costs of each type of management were related to clinical outcome in 134 patients with CLM. METHODS: The costs (both in terms of health care and to society) and benefits (treatment-added survival and normal quality of life survival) of chemotherapy treatment of 85 patients (HAI with implanted pump: 51 patients; and systemic chemotherapy: 34 patients) were compared with those in 49 patients managed by symptom control only. RESULTS: HAI chemotherapy cost the most (Pound Sterling 18,263 per patient) and symptom control the least (Pound sterling 2136 per patient). When survival was included, HAI was the most cost-effective treatment (health care cost per life year gained with HAI vs. systemic chemotherapy: Pound Sterling 24,604; systemic chemotherapy vs. symptom control: Pound Sterling 32,788), but there was no difference with regard to health care cost per normal quality of life gained. Societal costs incurred by lost work time and welfare payments during illness were higher for HAI (Pound Sterling 12,897) than systemic chemotherapy (Pound Sterling 9143) or symptom control (Pound Sterling 8090) because HAI-treated patients lived longer and, although working longer and contributing more productivity to society, lost more work days than other patients. CONCLUSIONS: The least expensive management for CLM was symptom control, whereas systemic and HAI chemotherapies were equally cost-effective in producing added normal quality survival for health care resources expended. Although overall societal costs were higher for HAI than for either systemic chemotherapy or symptom control, the cost benefit was difficult to interpret because of uncertain attitudes regarding continued work during terminal illness. PMID- 9731891 TI - Microsatellite instability and p53 mutations in sporadic right and left colon carcinoma: different clinical and molecular implications. AB - BACKGROUND: Left and right colon carcinomas can display different clinical, pathologic, and genetic characteristics. The purpose of this study was to characterize multiple molecular genetic alterations in sporadic colon carcinoma and to correlate them with the location of the tumors and with lymph node metastasis. METHODS: One hundred and twenty-five cases of sporadic colon carcinoma (50 in the right colon and 75 in the left colon in patients with no family history of colon carcinoma) were studied. Status of the p53 gene was assessed by polymerase chain reaction (PCR), single strand conformation polymorphism, and sequencing at exons 5-8. Microsatellite instability was analyzed with five microsatellite markers at chromosome 18. The mismatch repair genes hMLH1 and hMSH2 were studied in tumors found to have microsatellite instability by PCR and sequencing. RESULTS: In the 125 cases studied, there were 53 tumors with mutations of the p53 gene. p53 mutations correlated with lymph node metastases from right colon carcinoma cases (61%), and all cases with p53 mutations and microsatellite instability were AJCC/UICC Stage III (Dukes Stage C). In the right colon carcinoma cases the rate of microsatellite instability was related to the tumor size (19% in tumors measuring < 4 cm, and 34% in tumors measuring > 4 cm). No correlation between microsatellite instability and p53 mutations was detected. In the left colon carcinoma cases, p53 mutations were detected in 41% of tumors and microsatellite instability in 14%; neither finding was related to the tumor size. Mutations of the hMLH1 and hMSH2 mismatch repair genes were detected in 7 of 24 cases with marked microsatellite instability. CONCLUSIONS: Microsatellite instability is prone to occur in sporadic right colon carcinoma during tumor growth and is not associated significantly with mutations in the hMLH1 and hMSH2 mismatch repair genes or in the p53 gene. Concomitant detection of microsatellite instability and p53 mutations in right colon carcinoma is associated with the presence of lymph node metastases. PMID- 9731892 TI - Infrequent somatic mutation of the adenomatous polyposis coli gene in aberrant crypt foci of human colon tissue. AB - BACKGROUND: The authors examined somatic mutations of the adenomatous polyposis coli (APC) gene in 84 human aberrant crypt foci (ACF) to determine whether APC gene mutations were involved in the histologic progression of ACF. METHODS: Mutation cluster regions of the APC gene were subjected to polymerase chain reaction single-strand conformation polymorphism analysis and direct sequencing. RESULTS: Four kinds of deletion were detected in the mutation cluster regions of APC gene in five ACF. APC mutation was detected in 1 of 18 ACF with Stage I abnormalities (6%). Four of 10 adenomatous ACF (40%) harbored the mutation. There were no mutations in 56 hyperplastic ACF. CONCLUSIONS: These results suggest that APC mutations may be involved initially in only a limited number of adenomas in ACF. PMID- 9731893 TI - Retrospective analysis of the effect of interferon therapy on the clinical outcome of patients with viral cirrhosis. AB - BACKGROUND: Recent data suggest that interferon therapy (IFN) can reduce the risk of progression to hepatocellular carcinoma (HCC) in patients with hepatitis C virus (HCV)-related cirrhosis. METHODS: A cohort of 189 patients with Child's Stage A cirrhosis of viral etiology followed prospectively were analyzed retrospectively to assess the effects of IFN on the clinical course and development of HCC. RESULTS: During a mean follow-up of 71.5+/-23.6 months, 7.9% of 88 treated and 21.8% of 101 untreated patients showed worsening of the Child's disease stage (P < 0.01); 5.6% of treated and 26.7% of untreated patients developed HCC (P < 0.001); and 3.4% of treated and 19.8% of untreated patients died of liver disease or underwent orthotopic liver transplantation (OLT) (P < 0.005). Using Cox's regression analysis, no treatment with IFN, high bilirubin and alkaline phosphatase (ALP) levels, and low leukocyte counts and prothrombin activity (PT) were associated significantly with worsening of Child's disease stage; no treatment with IFN, long term disease, low albumin and PT, and high gamma-glutamyl transpeptidase (GGT) were related significantly to HCC development; and no treatment with IFN, low albumin and PT, and high GGT and ALP were associated significantly with reduced survival. After adjustment for independent risk factors identified by multivariate analysis, the estimated cumulative probability of worsening of cirrhosis (P < 0.05), development of HCC (P < 0.001), and death or OLT (P < 0.005) was significantly lower in IFN-treated patients compared with untreated patients. This beneficial effect of therapy was statistically evident only in HCV positive patients. CONCLUSIONS: These results support the hypothesis that IFN improves clinical outcomes and reduces progression to HCC in patients with HCV-related cirrhosis. These conclusions, based on retrospective data, should be confirmed prospective. PMID- 9731894 TI - Development of a new staging system for patients with recurrent laryngeal squamous cell carcinoma. AB - BACKGROUND: The management of recurrent head and neck squamous cell carcinoma can be challenging to both physician and patient. This is due at least in part to the lack of an accurate and clinically applicable staging system for these patients. The purposes of this study were to examine the survival patterns of patients presenting with recurrent laryngeal tumors, identify key factors affecting prognosis, and combine these factors to create a new staging system to predict survival. METHODS: The methods included a retrospective chart review of 473 patients with laryngeal squamous cell carcinoma who received their initial treatment at Washington University between 1980 and 1992. From this population, 126 patients (27%) who developed recurrence were identified. RESULTS: The overall 2-year survival rate was 40% (50 of 124 patients). Four variables affected survival: initial TNM stage, initial treatment, morphologic extent of recurrence, and treatment of recurrence. These variables were entered into a multivariate analysis to determine independent prognostic significance. Three variables were found to be statistically significant: TNM stage (chi-square = 4.6; P = 0.03), initial treatment (chi-square 14.3; P = 0.0002), and extent of recurrence (chi square = 19.4; P = 0.0001). The process of conjunctive consolidation was used to combine significant variables to create a new staging system for laryngeal recurrence. CONCLUSIONS: This staging system provides accurate estimates of prognosis, involves no new technology to implement, can aid both the patient and physician in planning treatment, and can be used in observational studies to assess the relative effectiveness of competing therapies. PMID- 9731895 TI - The prognostic significance of fluorodeoxyglucose positron emission tomography imaging for patients with nonsmall cell lung carcinoma. AB - BACKGROUND: A fundamental property of malignant tumors is increased glucose metabolism, which can be estimated by imaging the glucose analog fluorodeoxyglucose (FDG). The aim of this study was to determine whether FDG uptake in lung carcinoma, as measured on positron emission tomography (PET) imaging in patients with newly diagnosed bronchogenic carcinoma, has prognostic significance. METHODS: A retrospective review of all patients with a new diagnosis of nonsmall cell lung carcinoma and FDG-PET imaging was performed. Stage at presentation, cell type, tumor size, and survival data were recorded. For each patient, a standardized uptake ratio (SUR) was calculated for the primary lesion on PET and was correlated with clinical information to determine prognostic significance. RESULTS: One hundred fifty-five patients, with a mean age of 56 years, were enrolled. One hundred eighteen patients (76%) had lesions with SUR values <10 and had a median survival of 24.6 months (95% confidence interval [CI]: 20.9-41.1). Thirty-seven patients (24%) had tumor lesions with SUR values >10 and had a median survival of 11.4 months (95% CI: 9.3-19.4). An SUR >10 correlated with poorer survival (P = 0.0049). Patients with primary lesions >3 cm and an SUR > 10 had the worst prognosis, with a median survival of 5.7 months (95% CI: 4.0-13.1). Multivariate analysis demonstrated than an SUR > 10 provided prognostic information independent of the clinical stage and lesion size. CONCLUSIONS: On PET studies of patients with lung carcinoma, FDG uptake within the primary lesion correlates with survival. PMID- 9731896 TI - Multimodality treatment of primary lymphoepithelioma-like carcinoma of the lung. AB - BACKGROUND: Lymphoepithelioma-like carcinoma (LELC) of the lung occurs at a higher frequency in Asian compared with Western patients. Its association with Epstein-Barr virus varies among different ethnic groups. METHODS: Nine patients with primary LELC of the lung treated at a single institution with a multimodality approach comprised of surgery, chemotherapy, and radiotherapy are reported. Chemotherapy was comprised of cisplatin, 100 mg/m2, on Day 1 and 5 fluorouracil, 1 g/m2, on Days 2, 3, and 4. RESULTS: Five male and 4 female patients were treated over a 3-year period. Eight patients were non-smokers. Three patients had operable disease. Two of these patients received adjuvant radiotherapy or chemotherapy and remained free of recurrence at 18 and 20 months, respectively; 1 patient received no adjuvant treatment, and palliative chemotherapy was given for subsequent recurrent disease. Six patients had inoperable disease and received palliative chemotherapy +/- radiotherapy. Five patients had distant metastatic disease at presentation. Of the 7 patients who were evaluable for response to chemotherapy, 71.4% had a partial response and 28.6% had progressive disease. One patient who was evaluable for response to radiotherapy achieved a partial response. CONCLUSIONS: Primary LELC of the lung has a high rate of systemic metastasis and is highly chemosensitive. A multimodality approach to the management of this disease is recommended. PMID- 9731897 TI - Intracellular accumulation of thallium as a marker of cisplatin cytotoxicity in nonsmall cell lung carcinoma: an application of inductively coupled plasma mass spectrometry. AB - BACKGROUND: Thallium-201 (201Tl) scintigraphy has been used to detect malignant pulmonary disease. The mechanism of Tl influx in tumor cells is believed to be similar to that of cisplatin (CDDP) mediated by sodium- and potassium-activated adenosine triphosphatase (Na-K ATPase), and the Na-K ATPase activity may determine the cellular CDDP accumulation and sensitivity to CDDP. The objective of this study was to determine the accumulation of CDDP and Tl in vitro by using inductively coupled plasma mass spectrometry (ICP-MS), a new analytic technique for detecting ultra trace elements, and to evaluate the correlations between cellular CDDP and Tl accumulation, between CDDP 50% inhibitory concentration (IC50) values and cellular CDDP accumulation, and between CDDP IC50 values and cellular Tl accumulation. METHODS: Eight nonsmall cell lung carcinoma (NSCLC) cell lines were used (five adenocarcinomas and three squamous cell carcinomas). The cell lines were exposed to CDDP or Tl for 1 hour, and the resulting cellular accumulation of platinum and Tl was determined by ICP-MS. CDDP IC50 values were determined by a soluble tetrazolium/formazan assay. RESULTS: The authors were able to measure cellular CDDP and Tl accumulation precisely, and heterogeneity in the cellular accumulation of CDDP and Tl existed among the NSCLC cell lines. A significant inverse correlation was observed between CDDP IC50 values and the cellular accumulation of both CDDP and Tl. CONCLUSIONS: ICP-MS is suitable for the determination of cellular CDDP and Tl accumulation in NSCLC cell lines. Cellular Tl accumulation determined by ICP-MS may reflect CDDP cytotoxicity rather than cellular CDDP accumulation. PMID- 9731898 TI - Multimodality treatment of 128 patients with locally advanced breast carcinoma in the era of mammography screening using standard polychemotherapy with 5 fluorouracil, epirubicin, and cyclophosphamide: prognostic and therapeutic implications. AB - BACKGROUND: Locally advanced breast carcinoma is associated with a poor prognosis. With single treatment modalities, i.e., surgery and/or radiation therapy, results have been consistently dismal. However, several earlier reports have indicated improvement in survival with a combined modality approach, i.e., the utilization of systemic therapy. METHODS: Between 1991 and 1994, 128 patients with locally advanced noninflammatory or inflammatory breast carcinoma (LABC) were treated with a combined modality strategy consisting of 4-6 courses of preoperative 5-fluorouracil (600 mg/m2), epirubicin (60 mg/m2), and cyclophosphamide (600 mg/m2) (FEC) every 3 weeks, followed by modified radical mastectomy or sector resection with axillary dissection in combination with postoperative radiotherapy and concomitant cyclophosphamide (850 mg/m2). Postoperatively, 3-5 adjuvant courses of FEC therapy were given. Nine percent of the patients received preoperative radiotherapy because the FEC therapy was not sufficiently effective. One-third of the patients were given tamoxifen (20 or 40 mg daily) at the end of the multimodal therapy. RESULTS: Clinical responses were observed in 60% of the patients; 5% had complete responses (CR) and 55% had partial responses (PR). Stable disease (SD) was observed in 40%. No patient had progressive disease (PD) preoperatively. With a median follow-up of 37 months, the median disease free survival (DFS) and median overall survival (OS) were 29 and 54 months, respectively. The actuarial 5-year DFS and OS were 36% and 49%, respectively. The locoregional recurrence rate was 20%, and 53% of the patients experienced systemic relapse. Univariate analysis revealed a significant prognostic difference according to clinical stage of LABC in favor of less advanced stages. Clinical and biologic parameters linked to a significantly worse prognosis were the presence of inflammatory breast carcinoma and peau d'orange. There was a significant trend of worse prognosis for patients receiving below 60% and 75% of the intended dose intensity with reference to DFS and OS, respectively. CONCLUSIONS: Standard dose preoperative and postoperative FEC therapy combined with surgery and radiotherapy in the era of mammography screening seem to yield results comparable to those achieved with other conventional strategies in the treatment of unscreened populations. PMID- 9731899 TI - Prediction of axillary lymph node involvement of women with invasive breast carcinoma: a multivariate analysis. AB - BACKGROUND: The increasing use of systemic therapy for women with lymph node negative breast carcinoma and earlier stage of disease at mammographic detection raises questions regarding the need for routine axillary lymph node dissection. Predictive modeling for lymph node involvement may be one way to reduce the need for axillary lymph node dissection and its morbidity. METHODS: A multivariate analysis of 12 factors predictive of axillary lymph node involvement was conducted in a population-based cohort of 4312 women with invasive breast carcinoma diagnosed between January 1, 1993 and December 31, 1996. RESULTS: Clinical palpability, lymphatic or vascular invasion, lesion size, margin status, histology, and patient age were independent predictors of axillary lymph node involvement. The model correctly identified lymph node status in 76.6% of cases. Model accuracy and fit were equally high when applied to randomly selected halves of the study subjects. Approximately 32.0% of the patients in the study sample (1363/4312) were identified as having an extremely high (91%; n = 1102) or low (10%; n = 261) risk of lymph node involvement. In a second analysis, a clinically useable, three-variable model identified a very low risk group of patients (n = 147) with a 4.8% risk of lymph node metastasis and a high risk group of patients (n = 1008) with a 74.2% risk of lymph node metastasis. Greater than 90% of subjects in the high risk group received adjuvant systemic therapy even if they were lymph node negative pathologically. CONCLUSIONS: A clinically useable, three variable model employing tumor and lymph node palpability, size, and lymphatic or vascular invasion can identify women with invasive breast carcinoma in whom axillary lymph node dissection is very unlikely to alter recommendations regarding adjuvant systemic therapy. PMID- 9731900 TI - Prognostic impact of morphometric nuclear grade of endometrial carcinoma. AB - BACKGROUND: The identification of high risk patients with endometrial carcinoma is considered essential for individualized therapy to improve prognosis and avoid overtreatment. The goal of the current study was to investigate the prognostic value of nuclear morphometry, particularly for patients with localized endometrial carcinoma. METHODS: In a prospective study including 115 patients primarily treated for endometrial carcinoma at Haukeland University Hospital in Bergen, Norway between 1981-1990, data regarding clinical variables, histologic type, histologic grade, DNA index, estrogen and progesterone receptor concentration and nuclear morphometry were collected. The median follow-up period for the survivors was 9 years (range, 5-15 years). RESULTS: International Federation of Gynecology and Obstetrics (FIGO) stage, histologic type, histologic grade (World Health Organization), DNA index, progesterone receptor concentration, mean nuclear area, greatest and smallest nuclear diameter, nuclear perimeter, and standard deviation to mean nuclear area all influenced survival significantly in univariate analyses. In multivariate analyses, FIGO stage and morphometrically determined mean nuclear size were identified as independent prognostic factors, whereas histologic grade had borderline significance. Histologic type, DNA index, progesterone receptor concentration, and standard deviation of mean nuclear area were not significantly associated with prognosis. The nuclear perimeter was identified as the most powerful prognostic morphometric factor, and in a separate multivariate analysis that included patients with localized disease only, it was also an independent prognostic factor. This also was the case for the subgroup of patients with endometrioid carcinoma, adenoacanthoma, or adenosquamous carcinoma. CONCLUSIONS: Morphometric nuclear grade was a stronger prognostic factor than subjective histologic grade, histologic type, DNA index, and hormone receptor concentration in endometrial carcinoma patients, including those patients with localized disease. PMID- 9731901 TI - Histologic characterization of rat ovarian carcinoma induced by intraovarian insertion of a 7,12-dimethylbenz[a]anthracene-coated suture: common epithelial tumors of the ovary in rats? AB - BACKGROUND: The surface epithelium of the human ovary simulates the mullerian form in tumor formation. In experimental animals, however, such a phenomenon has not previously been observed. METHODS: A chemical carcinogen, 7,12 dimethylbenz[a]anthracene (DMBA), was heated, and the central portion of a 3-0 silk suture was immersed in the melted carcinogen. After vaginal cytology, the DMBA-coated silk was inserted into the ovaries of 40 Wistar strain rats age 7 weeks. The animals were sacrificed when a tumor mass became large enough to extend the abdominal wall. The experimental period lasted for 60 weeks. The tumor histology was compared with that in human counterparts and the mullerian derivatives in rats. RESULTS: In 19 rats, including 4 animals with cornified vaginal smears at the time of DMBA treatment, an ovarian carcinoma developed within 36 weeks. In 17 epithelial tumors, the neoplastic cells proliferated to form papillary or glandular structures, and the lining epithelium consisted of flattened or cuboidal cells and columnar cells, often with pseudostratified nuclei. Adenocarcinoma components transformed into squamous cells in two cases. The remaining two neoplasms were pure mesenchymal tumors with histology of a fibrosarcomatous tumor, and one of them contained heterologous malignant osteoid components. In the remaining 21 rats, including 5 with cornified smears, no tumors developed during the experimental period. CONCLUSIONS: The histologies of DMBA-induced rat ovarian carcinoma simulated the epithelia of rat ovarian surface, fallopian tube, endometrium, and uterine cervix. The results of this study suggest that the surface epithelium of rat ovary also simulates the mullerian form in tumor formation. PMID- 9731902 TI - A Gleason score of 7 predicts a worse outcome for prostate carcinoma patients treated with radiotherapy. AB - BACKGROUND: In most reported surgical series, prostate carcinoma patients with a Gleason score of 7 have had worse outcomes than those with other moderately differentiated cancers. Because of variations in reporting grade and grouping Gleason scores, radiation series have conflicting results. METHODS: Five hundred sixty-three men with clinical Stage T1-T3, N0 or Nx, M0 adenocarcinoma of the prostate and known pretreatment prostate specific (PSA) levels received external beam radiation only. The median pretreatment PSA was 10.3 ng/mL (range, 0.2-191 ng/mL). The median duration of follow-up was 42 months (range, 2-114 months). Survival without biochemical failure (bNED) was defined as PSA < or = 1.5 ng/mL and not rising when measured on two consecutive occasions. RESULTS: The 5-year rate of bNED control for all 563 patients was 62%. Increasing Gleason score predicted for decreased bNED control (78% for 2-4, 63% for 5-6, 37% for 7, and 33% for 8-10 at 5 years; P = 0.0001 for overall comparison). The bNED control rate for patients with a Gleason score of 7 was significantly less than the rate for those with Gleason 5-6 in both univariate (P = 0.0008) and multivariate (P = 0.0068) analysis. T classification by palpation, pretreatment PSA, and dose were also shown to be independent predictors of bNED control in multivariate analysis. CONCLUSIONS: Even after adjustment for other known prognostic factors, a Gleason score of 7 was associated with worse bNED control than Gleason scores of 2-4 and 5-6 among patients treated with external beam radiotherapy only for clinically localized prostate carcinoma. Patients with a Gleason score of 7 should not be lumped together with those who have a Gleason score of 5-6; they may instead benefit from more aggressive treatment strategies. PMID- 9731903 TI - Interphase cytogenetic study of preoperative core biopsies for the prediction of early serum prostate specific antigen recurrence after radical prostatectomy of clinically localized prostate carcinoma. AB - BACKGROUND: Current clinical and pathologic methods are imprecise in predicting the extent of prostate carcinoma progression. This study was conducted to evaluate the role of interphase cytogenetics (ICG) with chromosome enumeration probes relative to the role of conventional pathologic characteristics in the preoperative prediction of postoperative tumor classification and recurrence of elevated serum concentrations of prostate specific antigen (PSA). METHODS: The authors performed ICG with enumeration probes for chromosomes 7, 17, and X on 6 microm sections of core biopsies from 75 patients with clinically localized adenocarcinoma of the prostate. Results were compared with pathologic findings (tumor classification, volume, and status of surgical margins) of the corresponding radical prostatectomy specimens and data on serum PSA measurements taken for 72 patients during a follow-up period of up to 42 months (mean, 19.8 months). In addition to ICG, biopsies were reviewed for primary, secondary, highest, and combined Gleason grades, number and bilaterality of positive biopsies, amount of carcinoma (percentage and core length of infiltration), amount of high grade carcinoma, and perineural invasion. RESULTS: Chromosome categories were unevenly distributed among pathologic tumor classification groups (P = 0.0008) and between groups with positive and negative surgical margins (P = 0.0116). Tumors with aneusomic chromosome numbers had larger volumes than those with eusomy and tetrasomy (P = 0.0022). Kaplan-Meier analysis demonstrated that PSA recurrences (> or = 0.4 ng/mL) were more frequent and observed earlier in patients with detected chromosomal aneusomies than in those with eusomic and tetrasomic chromosome numbers (P < 0.0001). Cox regression analysis indicated that ICG was the most valuable independent factor in predicting PSA recurrences. CONCLUSIONS: The detection of numeric chromosomal aberrations in preoperative core biopsies is an adverse prognostic sign. Grading based on the quantification of genetic changes might prove useful in the prognostic stratification of patients with clinically localized prostate carcinoma. PMID- 9731904 TI - Ten-year disease free survival after transperineal sonography-guided iodine-125 brachytherapy with or without 45-gray external beam irradiation in the treatment of patients with clinically localized, low to high Gleason grade prostate carcinoma. AB - BACKGROUND: The authors report observed 10-year brachytherapy results in the treatment of 152 consecutive patients with clinically organ-confined prostate carcinoma. METHODS: One hundred and fifty-two consecutive patients with T1-T3, low to high Gleason grade, prostate carcinoma were treated between January 1987 and June 1988 at Northwest Hospital in Seattle, Washington. Their median age was 70 years (range, 53-92 years). Of these 152 patients, 98 (64%) received an iodine 125 implant alone (Group 1), and the remaining 54 patients (36%), who were judged to have a higher risk of extraprostatic extension, also were treated with 45 gray (Gy) of external beam irradiation to the pelvis (Group 2). No patient underwent lymph node sampling, and none received androgen ablation therapy. Multivariate regression and the Mann-Whitney rank sum test were used for statistical analysis. Preoperative patient data with associated success or failure outcomes at 10 years after treatment were used for training and validating a back-propagation neural network prediction program. RESULTS: The average preoperative prostate specific antigen (PSA) value, clinical stage, and Gleason grade were 11.0 ng/mL, T2, and 5, respectively. The median posttreatment follow-up was 119 months (range, 3-134 months). Overall survival 10 years after treatment was 65%. At last follow-up only 3 of the 152 patients (2%) had died of prostate carcinoma. Ninety-seven patients (64%) remained clinically and biochemically free of disease at 10 years of follow-up and had an average PSA value of 0.18 ng/mL (range, 0.01-0.5 ng/mL). In these patients a period of 42 months was required to reach the average PSA (0.5 ng/mL). The median to last PSA follow-up was 95 months (range, 3-134 months). Postoperative needle biopsies were negative in 56% of patients, positive in 15% of patients, and not available in 29% of patients. Only 6% of patients developed bone metastasis. At 10 years there was no statistically significant difference in treatment outcome between patients who received iodine-125 alone, and those who received iodine-125 with 45-Gy external beam irradiation (P = 0.08). Nevertheless, in these two groups preoperative PSA, stage, and Gleason grade were significantly different (P < 0.01). In the artificial neural network analysis, pretreatment serum PSA was the most accurate predictor of disease-free survival. CONCLUSIONS: Percutaneous prostate brachytherapy is a valid and efficient option for treating patients with clinically organ-confined, low to high Gleason grade, prostate carcinoma. Observed 10-year follow-up documents serum PSA levels superior to those reported in several published external beam irradiation series, and comparable to those published in a number of published radical prostatectomy series. PMID- 9731905 TI - Prognostic risk factors that identify patients with clinical stage I nonseminomatous germ cell tumors at low risk and high risk for metastasis. AB - BACKGROUND: The purpose of this study was to develop a reliable model to identify clinical Stage I nonseminomatous germ cell tumors (NSGCTs) associated with low risk or high risk for occult retroperitoneal metastasis, so that the model could be used to customize the therapeutic approach for patients with these tumors. The model was to be based on pathohistologic parameters and immunohistochemical expression of proliferation markers, proteases, and adhesion molecules in the primary tumor. METHODS: One hundred forty-nine patients with clinical Stage I NSGCTs underwent retroperitoneal lymphadenectomy and were included in the study. Three to five paraffin embedded, formalin fixed tissue blocks were available from each patient and were analyzed for the following histopathologic features associated with pathologic Stage I or II disease: the presence or absence of vascular invasion (VI), the presence or absence of tunic invasion, and the percentage of each histologic type present in the primary tumor. Immunohistochemical expression of MIB-1, p53, bcl-2, cathepsin D, and E-cadherin was evaluated using a semiquantitative scoring system. Statistical analysis was performed with univariate and multivariate logistic regression models. RESULTS: The percentage of embryonal carcinoma (%EC, P < 0.001) and the presence of VI (P < 0.0001) and tunic invasion (P < 0.002) were the most significant independent risk factors associated with pathologic Stage II disease. A combination of %EC and VI allowed correct prediction of final pathologic stage for 88% of clinical Stage I patients. Cutoff values including both variables identified the correct pathologic stage for 131 of 149 patients (88%). Less than 45% EC and the absence of VI correctly identified pathologic Stage I disease in 91.5% of patients; more than 80% EC and the presence of VI correctly predicted pathologic Stage II in 88%. In univariate analysis, only p53 (P < 0.03) and E-cadherin (P < 0.001) expression were significantly different in the embryonal carcinoma component of pathologic Stage I and II NSGCT. To evaluate prospectively the clinical utility of the new derived cutoff points, the data were applied to 10 consecutive patients with clinical Stage I NSGCT who underwent retroperitoneal lymphadenectomy; pathologic Stage I and II were correctly predicted for 5 of 6 Stage I and 4 of 4 Stage II patients, respectively. CONCLUSIONS: %EC and the presence or absence of VI appear to be reliable prognosticators to identify patients at high risk and low risk for occult retroperitoneal disease. In cases of clinical Stage I NSGCT, p53, bcl-2, MIB-1, cathepsin D, and E-cadherin did not appear to be of prognostic significance. The authors recommend that all patients with clinical Stage I NSGCT have their primary orchiectomy specimens evaluated for %EC and the presence of VI to determine their risk for occult retroperitoneal metastasis. PMID- 9731906 TI - Prospective multicenter study of thyroiscarcinoma treatment: initial analysis of staging and outcome. National Thyroid Cancer Treatment Cooperative Study Registry Group. AB - BACKGROUND: A novel prognostic staging classification encompassing all forms of thyroid carcinoma was created for the National Thyroid Cancer Treatment Cooperative Study (NTCTCS) Registry, with the goal of prospective validation and comparison with other available staging classifications. METHODS: Patient information was recorded prospectively from 14 institutions. Clinicopathologic staging was based on patient age at diagnosis, tumor histology, tumor size, intrathyroidal multifocality, extraglandular invasion, metastases, and tumor differentiation. RESULTS: Between 1987 and 1995, 1607 patients were registered. Approximately 43% of patients were classified as NTCTCS Stage I, 24% Stage II, 24% Stage III, and 9% Stage IV. Patients with follicular carcinoma were more likely to have "high risk" Stage III or IV disease than those with papillary carcinoma. Of 1562 patients for whom censored follow-up was available (median follow-up, 40 months), 78 died of thyroid carcinoma or complications of its treatment. Five-year product-limit patient disease specific survival was 99.8% for Stage I, 100% for Stage II, 91.9% for Stage III, and 48.9% for Stage IV (P < 0.0001). The frequency of remaining disease free also declined significantly with increasing stage (94.3% for Stage I, 93.1%for Stage II, 77.8% for Stage III, and 24.6% for Stage IV). The same patients also were staged applying six previously published classifications as appropriate for their tumor type. The predictive value of the NTCTCS Registry staging classification consistently was among the highest for disease specific mortality and for remaining disease free, regardless of the tumor type. CONCLUSIONS: The NTCTCS Registry staging classification provides a prospectively validated scheme for predicting short term prognosis for patients with thyroid carcinoma. PMID- 9731908 TI - The National Cancer Data Base report on cancer of the vagina. AB - BACKGROUND: This study was conducted to determine practice patterns in the management of vaginal malignancy. METHODS: The National Cancer Data Base (NCDB), a large central registry of hospital case data, was reviewed for the 10-year period 1985-1994 for patients registered with a primary diagnosis of vaginal cancer. Patients with a prior history of malignancy were excluded. RESULTS: Between 1985-1994 4885 cases of vaginal cancer were submitted to NCDB. More than 90% were epithelial neoplasia with approximately 25% of these in situ lesions only. Squamous carcinoma was more common as the age of the patient progressed. Adenocarcinomas represented nearly all the carcinomas in the group of patients age < 20 years and were observed less frequently with advanced age. Relative survival at 5 years was stage-related: Stage 0: 96%; Stage I: 73%; Stage II: 58%; and Stages III-IV: 36%. Melanoma had an extremely poor prognosis with a 5-year survival rate of only 14%. A significant number of sarcomas occurred in children for whom chemotherapy played a major role in treatment. Chemotherapy was used less frequently in the older patients. Survival was better in the younger patients (90% vs. 30% in the older patients). CONCLUSIONS: Although vaginal cancer is the rarest of genital malignancies, it appears that treatment and results from the NCDB reported from multiple institutions followed prescribed treatment guidelines. PMID- 9731907 TI - A study evaluating the efficacy and tolerability of tropisetron in combination with dexamethasone in the prevention of delayed platinum-induced nausea and emesis. AB - BACKGROUND: Chemotherapy-induced emesis is one of the most disturbing side effects of cancer therapy. Control of acute emesis has improved substantially during recent years, but control of delayed emesis and nausea remains a challenging problem. The role of 5-HT3 receptor antagonists in the treatment of delayed emesis is disputed. METHODS: Tropisetron, a highly specific 5-HT3 receptor antagonist, was compared (as an adjunct to dexamethasone) with placebo in a randomized, double blind, multicenter trial for the prevention of delayed emesis during platinum-containing chemotherapy. Three hundred chemotherapy-naive women with gynecologic malignancies were included. The cisplatin dose was in the range of 50-100 mg/m2. RESULTS: Acute emesis was prevented completely in 87% of patients and acute nausea in 77% of patients in the complete series. During the complete delayed period (Days 2-6), total control of emesis was achieved in 77% of the dexamethasone and tropisetron-treated patients and in 72% of the patients receiving dexamethasone and placebo (P = 0.2473). During the same period nausea was controlled completely in 42% of the dexamethasone and tropisetron group and in 41% of the dexamethasone and placebo group. On Day 3, complete protection from nausea was achieved in 65% of patients receiving tropisetron and in 51% of patients receiving placebo (P = 0.0304). Constipation occurred more frequently in the tropisetron group. CONCLUSIONS: Tropisetron added to dexamethasone improved control of delayed nausea on Day 3 compared with placebo. No significant differences were recorded regarding control of delayed emesis. PMID- 9731909 TI - The National Cancer Data Base report on Hodgkin's disease for 1985-1989 and 1990 1994. AB - BACKGROUND: A national survey of the management of Hodgkin's disease patients based on cases in the National Cancer Data Base (NCDB) provides a basis for evaluating the results of educational and therapeutic programs. These patients are believed to have been drawn from all nationalities, native and migrant, and were reported by hospital cancer registries throughout the United States, including large and small community hospitals, university and other teaching hospitals, military and Veterans Administration (VA) hospitals, and National Cancer Institute (NCI)-Designated Centers. METHODS: Data submitted voluntarily to the NCDB were used to determine trends in patterns of patient care across time. For the period 1985-1994, data from 35,033 patients with newly diagnosed Hodgkin's disease were analyzed and separated into two time periods, 1985-1989 and 1990-1994. RESULTS: Data were analyzed with respect to age, race, histology, stage, treatment, and survival. The majority of patients (83.6%) were white, the age group with the highest incidence was 20-29 years, and nodular sclerosis was the most common histologic type. Staging was reported as a combination of clinical and pathologic stage ("combined stage"). The number of cases of reported stage increased from 51.7% for the years 1985-1989 to 75.7% for the years 1990 1994. Radiation therapy was used primarily to treat patients in Stages I and II, although the overall use of radiotherapy declined by 10% in the later period. The overall observed 5-year survival rate was 83.2%, and the disease specific observed survival rate was 84.9%. Stage for stage, survival was better for younger patients and poorer for older patients. CONCLUSIONS: The survey reflects the actual management of Hodgkin's patients disease. The reported cases for 1994 represent 60.6% of the estimated occurrences for that year in the U.S. There has been a significant improvement in the frequency of use of the staging system. A continuing increase in survival for patients with Hodgkin's disease is occurring. This method of studying disease management provides a measure of educational efforts and guides to developmental research. PMID- 9731910 TI - Radiation-induced sarcoma--50 years later. PMID- 9731911 TI - Resection of hepatic and pulmonary metastases in patients with colorectal carcinoma. PMID- 9731912 TI - Undifferentiated carcinoma with osteoclast-like giant cells of the pancreas and periampullary region. PMID- 9731913 TI - Expression of cytokeratin 20 in urinary cytology of patients with bladder carcinoma. PMID- 9731914 TI - Hand-foot syndrome following prolonged infusion of high doses of vinorelbine. PMID- 9731915 TI - Cancer among spouses: review of 195 couples. PMID- 9731916 TI - Intensified therapy for infants with acute lymphoblastic leukemia: results from the Dana-Farber Cancer Institute Consortium. PMID- 9731917 TI - Report of a panel on the relationship between public exposure to pesticides and cancer. PMID- 9731918 TI - Randomized database studies: a new method to assess drugs' effectiveness? AB - The need to evaluate drugs' effects in real clinical practice is increasingly important. Randomized clinical trials (RCTs) and database analyses (DBA) are the two main methods to assess treatments effectiveness. RCTs remain the "gold standard" for comparing alternative treatments. However, they are conducted under strict, protocol-driven conditions that may limit their generalizability. Advantages of new high quality clinical databases, on the other hand, include the simple and economic access to large number and range of cases, and the ability to capture all aspects of actual medical practice. The main potential limitation of DBA is the potential for comparison bias due to the lack of randomization. Despite the efforts to design naturalistic trials and to use sophisticated statistical techniques to minimize selection bias, the inherent limitations of both methods (problems of external and internal validity, respectively) have not been completely solved. Thus, the actual challenge is the development of some new strategy capable of generating results with an acceptable balance between internal and external validity. As randomization is essential to minimize comparison bias, we point out the possibility to include randomization modules in computer-based patient records. The theoretical foundation of these "randomized database studies" is the simultaneous use of both experimental and observational methods in the assessment of drugs' effectiveness. The progressive standardization of clinical practice and the development and adoption of improved computer-based patient records could facilitate the use of this new research strategy. PMID- 9731919 TI - Outcome of pre-hospital antibiotic treatment of meningococcal disease. AB - OBJECTIVE: To assess the effect of pre-hospital antibiotic treatment given by general practitioners to patients with meningococcal disease. DESIGN: A 16-year population-based historical follow-up study based on referral letters and hospital records in the County of North Jutland, Denmark. SUBJECTS: 320 patients with meningococcal disease, of whom 302 were examined by a general practitioner before admission to hospital. MAIN OUTCOME MEASURES: Death. RESULTS: 44 patients (14.6%) were given antibiotic treatment by the referring general practitioner. Nine of these (20.5%) died, compared with 16 (6.2%) patients who did not receive pre-hospital antibiotic treatment. The presence of skin bleeding, petechiae, and impaired consciousness were strongly associated with case fatality. Even after adjustment for these variables the odds ratio (OR) for death in patients treated with antibiotics was high (OR = 3.2; 95% CI, 0.9-10.6). In the 15 patients with skin bleeding (ecchymoses, suggillations) the case fatality rate was 100% in patients treated with antibiotics, and 50% in patients who did not receive antibiotics before hospitalization. If skin bleeding was replaced in the models by the presence of disseminated intravascular coagulation on admission, the OR for death in patients with pre-hospital antibiotic treatment was 35.9 (95% CI, 2.9-441.8) in the presence of disseminated intravascular coagulation and 1.9 (95% CI, 0.2-19.5) in its absence. CONCLUSIONS: Pre-hospital treatment is mainly given to the most severe cases with expected high case fatality, and this confounding by indication was probably not fully adjusted for with the available data. The results contradict previous findings but provide reason to doubt the benefit of pre-hospital antibiotic treatment in patients with meningococcal disease. PMID- 9731920 TI - Time trends in the treatment of acute myocardial infarction in Switzerland from 1986 to 1993: do they reflect the advances in scientific evidence from clinical trials? AB - Three acute coronary care surveys (1986, 1990, and 1993) were conducted in the Swiss region of Vaud-Fribourg on all men aged 25 to 64 years hospitalized for a definite myocardial infarction (218, 224, and 167 cases). Nearly all patients received anticoagulants and nitrates. The proportion of patients treated increased significantly, between 1986 and 1990, for antiplatelet drugs (from 51% to 96%) and thrombolytics (from 9% to 44%) and, between 1990 and 1993, for beta blockers (from 57% to 78%) and angiotensin-converting enzyme inhibitors (from 26% to 43%). The use of calcium antagonists and antiarrhythmics dropped over time. Coronary arteriography and angioplasty were increasingly performed (53% and 18% in 1993), although progressively postponed in-hospital stay. The observed trends reflect a rapid translation of clinical trials into medical practice. However the use of thrombolytics could be raised further by shortening the hospitalization delay (median: 3 hours in 1993) and door-to-needle time (median: 47 minutes) which remained stable over time. PMID- 9731921 TI - Differences in risk factors for being either a hepatitis B carrier or anti hepatitis C+ in a hepatoma-hyperendemic area in rural Taiwan. AB - This is a study of the differences in the risk factors for being either hepatitis B surface antigen positive [HBsAg(+)] or antibody to hepatitis C virus positive [Anti-HCV(+)] in A-Lein, a rural area in southern Taiwan, an area which also has a high hepatoma mortality rate. Three hundred eighty-five patients age > or =40 years participated in hepatoma screening at the A-Lein Community Health Center during 1995. Those who were HBsAg(-) and anti-HCV(-) or had coinfection of HBsAg(+) and anti-HCV(+) were excluded, leaving 293 patients: 109 HBsAg(+) and 184 anti-HCV(+). The anti-HCV(+) patients had a lower socioeconomic status (as defined by level of education and type of occupation) and were older than HBsAg(+) patients (P < 0.05). Those with higher alanine aminotransferase levels (ALT) also had a higher anti-HCV(+) to HBsAg(+) odds ratio (OR), and a dose response relationship was found, P < 0.0001. Anti-HCV(+) patients were more likely than HBsAg(+) patients to have a spouse who shared the infection, OR = 5.11; 95% CI, 2.30-11.28. Anti-HCV(+) patients were more likely than HBsAg(+) patients to have had blood transfusions (OR = 2.66; 95% CI, 1.20-5.89), frequent medical injections (OR = 2.64; 95% CI, 1.62-4.31), or injections by non-licensed medical providers (OR = 1.91; 95% CI, 1.18-3.09). Multiple logistic regression analysis showed that the significant factors for anti-HCV(+) patients vs. HBsAg(+) patients are drinking habit (OR = 3.45; 95% CI, 1.02-11.60), age (OR = 6.33; 95% CI, 2.93-13.68), and frequent medical injections (OR = 2.88; 95% CI, 1.65-5.03). The transmission of hepatitis C in A-Lein is closely related to low socioeconomic status, age, alcohol abuse, spouses being anti-HCV(+), and frequent medical injections, especially from non-licensed medical providers, including both pharmacists and those with no medical licensing whatsoever. These nonlicensed medical providers sometimes reuse needles to save money, which is a likely route of infection. PMID- 9731923 TI - Procedures for estimating growth rates in thoracic aortic aneurysms. AB - Thoracic aortic aneurysms (TAAs) are potentially lethal medical conditions often requiring surgical intervention. Reliable information on TAA growth rates and associated risk factors is important for managing this challenging patient population. Unfortunately, a number of studies have employed questionable statistical methods, leading to biased and imprecise estimates. The present study describes these statistical problems in existing studies and delineates procedures for obtaining more reliable results. Using data from the Yale Center for Thoracic Aortic Disease, the study compares TAA growth rate estimates using conventional methods versus the recommended approach of instrumental variables (IV) estimation. The IV approach is designed to mitigate problems of measurement errors inherent in existing estimates of TAA growth. The results demonstrate that IV estimation yields more robust and precise estimates of TAA growth rates and risk factors for TAA growth. For example, the conventional approach yields TAA growth rates that fluctuate substantially-from 0.12 cm/yr to 0.90 cm/yr-depending on (1) the minimum serial follow-up period for patient inclusion in the study and (2) how subjects with negative measured growth rates are handled. In contrast, growth rate estimates using the IV approach are much more robust, ranging from 0.12 to 0.13 cm/yr. The 95% confidence intervals of estimated TAA growth are much more compact using the IV approach as well. We conclude that the IV estimation procedure yields more reliable estimates of TAA growth than does the conventional approach. PMID- 9731922 TI - Association of serum albumin concentration, serum ionized calcium concentration, and blood pressure in the Third National Health and Nutrition Examination Survey. AB - A few small studies of white persons have found a positive association between serum albumin and blood pressure. However, this association might be due to ionized calcium. No data on albumin or ionized calcium have appeared for African Americans or Hispanics, and few for women. To explore the association of serum albumin (g/L) and ionized calcium (mmol/L) with both systolic and diastolic blood pressure, data from the Third National Health and Nutrition Examination Survey, 1988-94, were analyzed. Results from multiple regressions, controlling for age, overweight, alcohol intake, hematocrit, pulse, antihypertensive medication, and smoking indicate that serum albumin is positively correlated (P < 0.01) to systolic and diastolic blood pressure among non-Hispanic white men 25-59 and 60 89 years old. Ionized calcium was associated negatively with diastolic blood pressure among younger Mexican-American men. In this national sample, serum albumin was consistently associated with systolic and diastolic blood pressure only among non-Hispanic white men. PMID- 9731924 TI - Comparison of the Quality of Well-being Scale and the SF-36 results among two samples of ill adults: AIDS and other illnesses. AB - PURPOSE: To compare results of using the SF-36 Short Form 36 (SF-36) and the Quality of Well-being Scale (QWB) in characterizing health outcomes over time in patients having serious illnesses, including cancer and AIDS. BACKGROUND: The SF 36 and the QWB are alternative measures of health-related quality of life. The SF 36 is a morbidity measure that features a profile of nine dimensions. The QWB is a preference-based measure that combines morbidity and mortality into a single number. However, the QWB can also be scored and used as a profile. We compare SF 36 and QWB scores with different scoring methods to assess validity and sensitivity to change over time in health outcomes for adult patients with HIV infection, cancer, and other serious illnesses. SUBJECTS: 201 adults with serious illnesses, including 99 with AIDS and 102 with cancer or other illnesses. PROCEDURE: All subjects received both measures at baseline and at 6-month intervals thereafter, over a period of 21/2 years. RESULTS: In the profile mode, the QWB captured outcomes that characterize the AIDS syndrome. The SF-36 differentiated between the AIDS and other illnesses patients on some scales, but without consistent direction. However, the overall QWB showed a decrease in quality of life over time for both the AIDS and other illnesses patients while the SF-36 did not. This is because many patients died and these were counted as outcomes by the QWB and as missing data by the SF-36. CONCLUSIONS: The QWB appears to be better able to capture outcomes of serious illness over time than does the SF-36. PMID- 9731925 TI - Approximate standard errors and confidence intervals for indices of positive and negative agreement. AB - Indices of positive and negative agreement for observer reliability studies, in which neither observer can be regarded as the standard, have been proposed. In this article, it is demonstrated by means of an example and a small simulation study that a recently published method for constructing confidence intervals for these indices leads to intervals that are too wide. Appropriate asymptotic (i.e., large sample) variance estimates and confidence intervals for the positive and negative agreement indices are presented and compared with bootstrap confidence intervals. We also discuss an alternative method of interval estimation motivated from a Bayesian viewpoint. The asymptotic intervals performed adequately for sample sizes of 200 or more. For smaller samples, alternative confidence intervals such as bootstrap intervals or Bayesian intervals should be considered. PMID- 9731926 TI - Impact of an educational program on antibiotic use in a tertiary care hospital in a developing country. AB - A multi-cross-sectional study was conducted in a 2000-bed tertiary care university hospital in Bangkok, Thailand, from September 1993 to May 1994 to assess the effectiveness of an educational program on the use of antibiotics. Data on the study covered antibiotic usage both in-patients and out-patients. Data were collected for a 24-hour period every 2 weeks for 7 days for each 3 month period. The target population were residents, general practitioners, and sixth-year medical students. The educational program provided information derived from the data of inappropriate use of antibiotics during the pre-intervention period and guidelines on the use of antibiotics which were agreed to by a consensus among the faculty in all clinical departments. The study revealed: (1) the prevalence of antibiotic use and the cost of antibiotics during post intervention period was significantly decreased by 20%; (2) the use of antibiotic prophylaxis for obstetrics patients and patients undergoing cataract surgery decreased significantly; (3) there was a shift from second or third generation cephalosporins to cefazolin for surgical prophylaxis; (4) the duration of perioperative antibiotic prophylaxis was reduced to under 2 days; (5) there was a shift from netilmicin or amikacin to gentamicin for the treatment of community acquired infection; and (6) the mortality, median length of hospital stay, and nosocomial infection rate among the patients who received antibiotics during the post-intervention period were not significantly different from those during the pre-intervention period. These results suggest that this educational program comprising information feedback and antibiotic usage guidelines was effective in improving antibiotic use at this tertiary care university hospital in Thailand. PMID- 9731927 TI - A national HIV community cohort: design, baseline, and follow-up of the AmFAR Observational Database. American Foundation for AIDS Research Community-Based Clinical Trials Network. AB - This article describes the design, methodology, baseline distributions, and general follow-up characteristics of the American Foundation for AIDS Research (AmFAR) National Observational Database (ODB) Project including the benefits and limitations of collecting information on a large simple cohort in the HIV community setting. The study prospectively followed 15,611 HIV-positive men and women and collected longitudinal and cross-sectional data on demographics, medical conditions, drug therapies, laboratory parameters, and survival. Participants were followed between October 1990 and December 1993 by 252 community-based sites coordinated by 22 centers in the Community-Based Clinical Trials Network (CBCT Network) throughout the United States (including Puerto Rico) and Toronto, Canada. The ODB provided quantitative information on a national level needed to track the HIV epidemic and plan clinical trials conducted through the Network, and to provide sites with local databases to monitor patients and facilitate access to therapies in clinical trials. Overall, the ODB contains information on 1,925 women (12%) and 13,686 men (88%), 60% white, 20% African American, 17% Latino/Hispanic, with 56,254 baseline and follow up forms, a median follow-up of about 12 months, a 16% loss-to-follow-up, and an 11% mortality rate. AmFAR plans to place the ODB in the public domain. PMID- 9731928 TI - Differences in perceived and presented adverse drug reactions in general practice. AB - BACKGROUND: Postmarketing surveillance (PMS) studies are frequently based on data from general practitioners (GPs). Patients, however, do not always report to their GP suspected adverse drug reactions. SETTING: A postmarketing cohort study on adverse reactions to sumatriptan, performed with assistance of drug dispensing GPs in The Netherlands. METHODS: Questionnaires were sent to all drug-dispensing GPs in The Netherlands, as well as to their patients on sumatriptan. To avoid bias, no specific adverse reactions were mentioned in the questionnaires. RESULTS: Of the GPs, 589 (86%) responded; of the patients, 1202 (70%) responded. The most frequently reported suspected adverse reactions to sumatriptan reported by the GPs were dizziness (1.7%), nausea or vomiting (1.5%), drowsiness or sedation (1.4%), and chest pain (1.3%). The most frequently reported suspected adverse reactions by the patients were paraesthesia (11.7%), dizziness (8.1%), feeling of heaviness (8.0%), and chest pain (7.9%). Neither the GPs nor the patients reported serious adverse reactions. CONCLUSIONS: First, patients experience significantly more suspected adverse reactions than are registered by their GP. In view of this higher frequency of reporting of suspected adverse reactions, postmarketing studies with data from GPs only, may underestimate the cumulative incidence of adverse reactions. Second, we conclude that it is possible to obtain useful additional information about adverse drug reactions from patients by sending them questionnaires via their GP. PMID- 9731930 TI - QEEG in migraine without aura. PMID- 9731929 TI - Sensitivity to light and noise in tension-type and cervicogenic headache. PMID- 9731931 TI - Endothelin in migraine patients. PMID- 9731932 TI - Migraine in childhood and adolescence. PMID- 9731933 TI - Differential effects of migraine drugs on cerebral blood flow autoregulation. AB - The effect of the migraine drugs ergotamine and sumatriptan on the cerebral blood flow (CBF) autoregulation was studied in halothane/nitrous oxide-anesthetized normotensive Wistar Kyoto rats. Ergotamine, an ergot alkaloid affecting 5HT, norepinephrine, and dopamine receptors, was administered intravenously as a single dose of 25 microg/kg. Sumatriptan, a selective 5HT1-like receptor agonist, was administered by intravenous infusion of 300 microg/kg/h. CBF was measured with the intracarotid 133Xe-injection method. The blood pressure limits of CBF autoregulation were determined by computerized least sum of square analysis. CBF autoregulation was preserved after both ergotamine and sumatriptan. Ergotamine shifted the lower blood pressure limit of CBF autoregulation towards higher blood pressures from 60 +/- 3 mmHg to 82 +/- 4 mmHg (p<0.01), but did not significantly affect the upper blood pressure limit of CBF autoregulation. Sumatriptan had no significant effects on the blood pressure limits of CBF autoregulation. PMID- 9731934 TI - Photophobia and phonophobia in tension-type and cervicogenic headache. AB - Light and sound-induced discomfort and pain thresholds were measured in 26 patients with cervicogenic headache, in 40 patients with tension-type headache, and in 100 headache-free controls. Neither headache group was significantly different as to photophobia and phonophobia, but both were significantly more sensitive to light and sound than controls (p<0.0001), even when patients were tested in the headache-free period (p<0.05). Episodic and chronic tension-type headache had similar photo- and phonophobia thresholds (p> or =0.7). Tension-type headache patients were more photo- and phonophobic during headache than outside attack (p<0.05), but this was not true for cervicogenic headache (p> or =0.56). In cervicogenic headache patients, photophobia (p<0.05) but not phonophobia (p=0.28) was greater on the symptomatic side than on the non-symptomatic side. PMID- 9731935 TI - Pattern reversal visual evoked potentials in migraine with aura and migraine aura without headache. AB - Pattern reversal visual evoked potentials (PVEPs) were recorded in 20 patients with migraine with aura (MA), 19 patients with migraine without headache (migraine equivalent; ME) during interictal periods, and 34 normal subjects. All migraine patients had hemianopsia or fortification spectra during attacks. In both MA and ME patients of less than 49 years of age, there were significant (p<0.01) differences in amplitude of PVEPs at the mid-occipital and contralateral to visual aura electrode sites compared to normal subjects. Amplitude of PVEPs in MA and ME showed significant (p<0.001) increases when recorded soon after attacks, especially within 10 days. There was a significant (p<0.01) correlation between percentage asymmetries and the duration of illness in both MA and ME. We conclude from our PVEP findings that cortical spreading depression remains the most likely explanation for the migraine visual aura. PMID- 9731936 TI - EEG spectral analysis in migraine without aura attacks. AB - In 16 patients suffering from migraine without aura, we examined quantitative EEG and steady-state visual evoked potentials (SSVEPs) at 27 Hz stimulation during the critical phase of migraine and in attack-free periods. The main spontaneous EEG abnormalities found during the critical phase were the slowing and asymmetry of the dominant frequency in the alpha range. The amplitude of the SSVEP F1 component was significantly reduced during the attack phase compared with the intercritical phase; in the latter condition the visual reactivity to 27 Hz stimulus was increased over almost the entire scalp compared with normal subjects. The EEG abnormalities confirm a fluctuating modification of alpha activity during the migraine attack, probably related to a functional disorder. The suppression of visual reactivity during the migraine attack could be related to a phenomenon of neuronal depolarization such as spreading depression, occurring in a situation of central neuronal increased excitability predisposing to migraine attacks. PMID- 9731937 TI - Endothelin in migraine patients. AB - Endothelins (ETs) are the most potent vasoconstrictors known, and may be the mediators of the vasoconstrictive phase in migraine attacks. We studied 31 previously selected migraine patients with (9) and without (22) aura ictally and interictally to determine their plasma ET-1 values. The mean interictal and ictal values were 5.3 pg/ml (SD 1.8) and 6.4 pg/ml (SD 3.9), respectively. The ictal values were markedly elevated at the beginning of the migraine attack (<2 h) and declined to interictal or even lower level later (4 to 6 h) in the course of an attack. The local vasoconstriction at the beginning of a migraine attack might be ET-mediated secondarily to serotonin activation. PMID- 9731938 TI - Migraine in childhood and adolescence. A critical study of the diagnostic criteria and of the influence of age on clinical findings. AB - We studied 253 children aged <15 years. Phase 1 included 193 children with migraine (1.1 and 1.2) divided into two groups (<10 and > or = 10 years). We studied the relationship between age and migraine type, headache characteristics, and associated symptoms of the International Headache Society (IHS) definition. A higher frequency of migraine with aura, pulsatile quality, and unilateral location was observed in older children. In phase 2 we studied 176 children with headache (excluding migraine with aura), comparing diagnostic criteria, definition items, sensitivity, and specificity. The results showed that item B of the definition was the most frequent cause of exclusion in the 1.7 diagnostic group. Compared with Vahlquist and the IHS, the Prensky criteria were the most sensitive. Sensitivity was >70% for pain of moderate/severe intensity, duration between 2 and 48 h, isolated photophobia, isolated phonophobia, and aggravation with physical activity. Specificity was >70% for nausea, vomiting, phonophobia and photophobia, isolated photophobia, aggravation with physical activity, and isolated phonophobia. Based on three alternative definitions, each modifying one item of the IHS definition, the sensitivity and specificity of these alternative definitions were compared with the "extended" criteria (children with migraine without aura and migrainous disturbance, according to the IHS criteria, grouped together). Exclusion of headache duration increased sensitivity by 10%, compared to restrictive IHS criteria, without decreasing specificity. PMID- 9731939 TI - Screening for migraine in the general population: validation of a simple questionnaire. AB - We present validation of a simple questionnaire designed to screen the general population for migraine. It comprises four questions: (1) "Have you ever had migraine?" (2) "Have you ever had severe headache accompanied by nausea?" (3) "Have you ever had severe headache accompanied by hypersensitivity to sound and light?" (4) "Have you ever had visual disturbances lasting 5-60 min followed by headache?" A telephone interview carried out by a physician was used as an index of validity. The study population was 5,360 twins from the population-based Danish Twin Registry. All twin pairs where at least one twin had answered "yes" to at least one of our questions were eligible for the telephone interview (n=2,272 twins). The response rate to the questionnaire was 87%; the participation rate in the telephone interview was 90%. The questionnaire ascertained 85% of all migraineurs (sensitivity). A combination of two questions (questions 1 and 4) extracted 93% of the twins with migraine with aura and 74% of the twins with migraine without aura, yet only 26% of the sample needed to be interviewed. We conclude that in the Danish population two simple questions are sufficient to screen for migraine in selecting participants for a diagnostic clinical interview. Our questionnaire clearly merits further study to document its universal applicability. PMID- 9731940 TI - Flunarizine in migraine prophylaxis: predictive factors for a positive response. AB - The efficacy of flunarizine in migraine prophylaxis is confirmed in both open and controlled trials. However, it is unknown what factors may influence a good response to prophylaxis with flunarizine. The aim of this study was to determine the possible predictive factors for therapeutic responsiveness to 3 months' treatment with flunarizine. One-hundred headache patients treated with flunarizine were evaluated. We considered "responders" those patients who recorded a reduction in migraine frequency of 75% after treatment. Statistical analysis revealed four factors which might influence therapeutic responsiveness in our patients. Positive factors were a family history (p<0.01) and high intensity of pain (p<0.01); negative factors were frequent attacks (p<0.01) and a history of analgesic abuse (p<0.001). Patients with no previous history of analgesic abuse, low frequency of attacks at baseline, higher levels of migraine pain, and positive family history constitute the prototype of flunarizine long term treatment responders. PMID- 9731941 TI - No effect of prophylactic treatment with metoprolol on cerebrovascular CO2 reactivity in migraineurs. AB - The prophylactic effect of metoprolol in the treatment of migraine is well known, but its mode of action is still unclear. In the past, increased CO2 reactivity has been reported as one pathognomic finding in interictal migraineurs. Using transcranial Doppler we assessed CO2 reactivity in 20 migraineurs before and 3 h after the first intake of 50 mg metoprolol, and subsequently twice after 1 and 8 weeks of continuous therapy with 150 mg metoprolol/d. Before initiation of therapy, migraineurs as a group had increased CO2 reactivity (p=0.07) compared to 20 age- and sex-matched volunteers. While treatment with metoprolol has been reported to affect amplitudes of increased contingent negative variation or visual evoked potentials in interictal migraineurs, it had no influence on enhanced CO2 reactivity in the present study. Moreover, the pretreatment value of CO2 reactivity did not correlate with the clinical efficacy of metoprolol after a 2-month treatment period. PMID- 9731942 TI - Trigeminal-autonomic cephalgias (TACs) PMID- 9731943 TI - Hypnic headache syndrome. PMID- 9731944 TI - Olfaction in migraine. PMID- 9731945 TI - Sumatriptan-induced axial dystonia in a patient with cluster headache. PMID- 9731946 TI - Bioavailability of spiramycin and lincomycin after oral administration to fed and fasted pigs. AB - The disposition of spiramycin and lincomycin was measured after intravenous (i.v.) and oral (p.o.) administration to pigs. Twelve healthy pigs (six for each compound) weighing 16-43 kg received a dose of 10 mg/kg intravenously, and 55 mg/kg (spiramycin) or 33 mg/kg (lincomycin) orally in both a fasted and a fed condition in a three-way cross-over design. Spiramycin was detectable in plasma up to 30 h after intravenous and oral administration to both fasted and fed pigs, whereas lincomycin was detected for only 12 h after intravenous administration and up to 15 h after oral administration. The volume of distribution was 5.6 +/- 1.5 and 1.1 +/- 0.2 L/kg body weight for spiramycin and lincomycin, respectively. For both compounds the bioavailability was strongly dependent on the presence of food in the gastrointestinal tract. For spiramycin the bioavailability was determined to be 60% and 24% in fasted and fed pigs, respectively, whereas the corresponding figures for lincomycin were 73% and 41%. The maximum plasma concentration of spiramycin (Cmax) was estimated to be 5 microg/mL in fasted pigs and 1 microg/mL only in fed pigs. It is concluded that an oral dose of 55 mg/kg body weight is not enough to give a therapeutically effective plasma concentration of spiramycin against species of Mycoplasma, Streptococcus, Staphylococcus and Pasteurella multocida. The maximum plasma concentration of lincomycin was estimated to be 8 microg/mL in fasted pigs and 5 microg/mL in fed pigs, but as the minimum inhibitory concentration for lincomycin against Actinobacillus pleuropneumoniae and P. multocida is higher than 32 microg/mL a therapeutically effective plasma concentration could not be obtained following oral administration of the drug. For Mycoplasma the MIC90 is below 1 microg/mL and a therapeutically effective plasma concentration of lincomycin was thus obtained after oral administration to both fed and fasted pigs. PMID- 9731947 TI - Intracellular accumulation, subcellular distribution and efflux of tilmicosin in swine phagocytes. AB - Tilmicosin is a semi-synthetic macrolide antibiotic, currently approved for veterinary use in cattle and swine respiratory disease. As the concentrations of tilmicosin are generally low in swine lung tissue, the interaction of tilmicosin with three types of swine phagocytes (monocyte-macrophages, alveolar macrophages, and neutrophils) was evaluated to provide an understanding of clinical efficacy. After incubation with radiolabelled tilmicosin, uptake was determined and expressed as the ratio of the intracellular (Ci) to the extracellular (Ce) drug concentration (Ci/Ce). Tilmicosin was avidly accumulated by the swine phagocytes (Ci/Ce 48-69 at 4 h incubation) with 51 to 85% localized in the lysosomes. Uptake was dependent on cell viability, temperature and pH, but was not influenced by the metabolic inhibitors, sodium cyanide or potassium fluoride. However, lipopolysaccharide (LPS) exposure increased tilmicosin uptake by the swine phagocytes. In neutrophils, upon removal of extracellular tilmicosin, 60% of the intracellular tilmicosin was effluxed within the first 30 min, but after 4 h of incubation in drug-free medium, 25% remained cell-associated. In contrast, after 4 h of incubation in drug-free medium, 60% and 45% of tilmicosin remained cell associated, within alveolar macrophages and monocyte-derived macrophages, respectively. Tilmicosin uptake was observed to increase lysosomal enzyme (acid phosphatase, lysozyme and beta-glucuronidase) production. Finally, neutrophils were shown to transport and efflux bioactive tilmicosin in a test system measuring both neutrophil chemotaxis under agarose and a bioassay measuring inhibition of bacterial growth in the presence of antibiotic in agar. These in vitro interactions of tilmicosin with swine phagocytes suggest an integral role in effecting clinical efficacy. PMID- 9731948 TI - Disposition of mirosamicin in honeybee hives. AB - Disposition of mirosamicin, a macrolide antibiotic, to honeybee adults, larvae, honey and royal jelly in the beehive after in-feed administration to adult bees was studied. Treatment was initiated at the end of July when the availability of natural pollen and nectar was poor. The drug was mixed with pollen-substitute paste and administered to honeybee colonies continuously for a week at a dosage of 200 mg/hive/week. High distributions in adult bees, jelly, larvae and a relatively low distribution in honey, of mirosamicin were observed. One day dosing of microsamin in sucrose syrup, a nectar substitute, resulted in a very high and long lasting residue in honey. Both royal and worker jelly, secreted from the jelly glands of adult bees, are acidic, so that a high distribution of a basic drug, such as mirosamicin, in jelly can be expected. This mechanism was considered to be responsible for a high concentration of mirosamicin in honeybee larvae, the host of Paenibacillus-larvae infection (American foulbrood), as primary larval food is jelly. PMID- 9731949 TI - Antibiotic effects on cytochromes P450 content and mixed-function oxygenase (MFO) activities in the American alligator, Alligator mississippiensis. AB - There are no Food and Drug Administration (FDA)-approved antimicrobial agents for use in cultured American alligators (Alligator mississippiensis) destined for human consumption yet some producers administer antibiotics for prophylaxis. The cytochromes P450-dependent mixed-function oxygenases (MFO) catalyze the oxidation of xenobiotic compounds such as drugs, pesticides and polycyclic aromatic hydrocarbons. Herein, we describe the effects of oxytetracycline, ceftazidime and enrofloxacin on the MFO system of the American alligator, Alligator mississippiensis. Juvenile alligators (4 animals/treatment) were administered these antibiotics intraperitoneally in an effort to induce hepatic microsomal cytochromes P450. Alligators treated with enrofloxacin exhibited emesis and convulsive spasms within 5 min of the initial injection. Total hepatic cytochromes P450 contents were significantly decreased in oxytetracycline-and enrofloxacin-pretreated alligators. In vitro hepatic microsomal benzyloxyresorufin O-dealkylase (BROD) activity was significantly decreased by enrofloxacin pretreatment. Western blots of proteins from antibiotic-pretreated alligator hepatic microsomes incubated with several mammalian and fish cytochromes P450 (CYP) antibodies exhibited little or no induction of CYP1A1, 2B, 2C and 2E1. In vitro incubation with enrofloxacin and oxytetracycline caused a concentration-dependent decrease in alkyl-substituted phenoxazone dealkylase activities catalyzed by phenobarbital- and 3-methylcholanthrene-induced alligator hepatic microsomes. PMID- 9731950 TI - Adrenocortical and metabolic responses to dobutamine infusion during halothane anaesthesia in ponies. AB - The study investigated whether hypotension in halothane-anaesthetised ponies is the stimulus inducing an endocrine stress response by assessing the effect of maintenance of normotension with a dobutamine infusion. Groups of six ponies were studied. After premedication with acepromazine (0.04 mg/kg) anaesthesia was induced with thiopentone (10 mg/kg) and maintained for 120 min with halothane (group AN). Dobutamine was infused to effect (1.1-4.4 microg/kg/min) to maintain arterial pressure at pre anaesthetic levels. The conscious group (CON) were prepared as for AN and then received only dobutamine infusion 1.0 microg/kg/min for 120 min. Arterial blood pressure, pH, oxygen and carbon dioxide tension, pulse rate, haematocrit, and plasma cortisol, glucose and lactate concentrations were measured before, at 20 min intervals during anaesthesia, and 20 and 120 min after anaesthesia ceased. Blood pressure remained close to control in both groups. The AN group became hypercapnic and acidotic, pulse rate and haematocrit increased, cortisol increased more than twofold and plasma glucose and lactate did not change. All values remained at control in the CON group except for small increases in haematocrit and decreases in pulse rate. Maintenance of normotension during halothane anaesthesia did not blunt the adrenocortical response to anaesthesia nor did the same dose of dobutamine alone increase plasma cortisol. Hypotension appears not to be the sole stimulus to equine adrenocortical activity during halothane anaesthesia. PMID- 9731951 TI - Pharmacokinetics of enrofloxacin after intravenous, intramuscular and oral administration in houbara bustard (Chlamydotis undulata macqueenii). AB - The in-vitro activity of enrofloxacin against 117 strains of bacteria isolated from bustards was determined. Minimum inhibitory concentrations for 72% of the Proteus spp., E. coli, Salmonella spp. and Klebsiella spp. (n = 61) and for 48% of the Streptococci spp. and Staphylococci spp. (n = 31) were < or = 0.5 microg/mL. The minimum inhibitory concentration (MIC) of 76% of Pseudomonas spp. (n = 25) was < or = 2 microg/mL. Fourteen strains were resistant to concentrations > or = 128 microg/mL. The elimination half-lives (t1/2 elim beta) (mean +/- SEM) of 10 mg/kg enrofloxacin in eight houbara bustards (Chlamydotis undulata) were 6.80 +/- 0.79, 6.39 +/- 1.49 and 5.63 +/- 0.54 h after oral (p.o.), intramuscular (i.m.) and intravenous (i.v.) administration, respectively. Enrofloxacin was rapidly absorbed from the bustard gastro-intestinal tract and maximum plasma concentrations of 1.84 +/- 0.16 microg/mL were achieved after 0.66 +/- 0.05 h. Maximum plasma concentration after i.m. administration of 10 mg/kg was 2.75 +/- 0.11 microg/mL at 1.72 +/- 0.19 h. Maximum plasma concentration after i.m. administration of 15 mg/kg in two birds was 4.86 microg/mL. Bioavailability was 97.3 +/- 13.7% and 62.7 +/- 11.1% after i.m. and oral administration, respectively. Plasma concentrations of enrofloxacin > or = 0.5 microg/mL were maintained for at least 12 h for all routes at 10 mg/kg and for 24 h after i.m. administration at 15 mg/kg. Plasma enrofloxacin concentrations were monitored during the first 3 days of treatment in five houbara bustards and kori bustards (Ardeotis kori) with bacterial infections receiving a single daily i.m. injection of 10 mg/kg for 3 days. The mean plasma enrofloxacin concentrations in the clinical cases at 27 and 51 h (3.69 and 3.86 microg/mL) and at 48 h (0.70 microg/mL) were significantly higher compared with the 3 h and 24 h time intervals from clinically normal birds. The maximum plasma concentration (Cmax)/MIC ratio was ranked i.v. (10/mg/kg) > i.m. (15 mg/kg) > i.m. (10 mg/kg) > oral (10 mg/kg), but it was only higher than 8:1 for i.v. and i.m. administrations of enrofloxacin at 10 mg/kg and 15 mg/kg, respectively, against a low MIC (0.5 microg/mL). A dosage regimen of 10 mg/kg repeated every 12 h, or 15 mg/kg repeated every 24 h, would be expected to give blood concentrations above 0. 5 microg/mL and hence provide therapeutic response in the bustard against a wide range of bacterial infections. PMID- 9731952 TI - The pharmacokinetics of furosemide in anaesthetized horses after bilateral ureteral ligation. AB - The pharmacokinetics of furosemide were investigated in anaesthetized horses with bilateral ureteral ligation (BUL) with (n = 5) or without (n = 5) premedication with phenylbutazone. Horses were administered an intravenous (i.v.) bolus dose of furosemide (1 mg/kg) approximately 60-90 min after BUL. Plasma samples collected up to 3 h after drug administration were analysed by a validated high performance liquid chromatography method. Median plasma clearance (CLp) of furosemide in anaesthetized horses with BUL was 1.4 mL/min/kg. Apparent steady state volume of distribution (Vd(ss)) ranged from 169 to 880 mL/kg and the elimination half life (t1/2) ranged from 83 min to 209 h. No differences in plasma concentration or kinetic parameter estimates were observed when phenylbutazone was administered before furosemide administration. BUL markedly reduces the elimination of furosemide in horses and models the potential effects that severe changes in kidney function may have on drug kinetics in horses. PMID- 9731953 TI - Femtomolar bradykinin-induced relaxation of isolated bovine coronary arteries, mediated by endothelium-derived nitric oxide. AB - We reported previously that bradykinin induces endothelium-dependent relaxation at nanomolar (nM) concentrations in isolated bovine coronary arteries with an intact endothelium. Recently we have found that in the presence of 10 microM indomethacin, femtomolar (fM) concentrations of bradykinin induce endothelium dependent relaxation in some bovine coronary arteries (approximately 10% of the coronary arteries examined). The present study was designed to characterize the relaxation induced by fM bradykinin. Relaxation was completely abolished by repeated application of fM bradykinin, by 100 microM N(omega)-nitro-L-arginine methyl ester and by 10 microM methylene blue. Relaxation induced by nM bradykinin was partly affected by these treatments. Relaxation induced by both concentrations of bradykinin was inhibited by a B2-kinin receptor antagonist, [Thi5,8, D-Phe7]-bradykinin, in a concentration-dependent manner, but not by a B1 kinin receptor antagonist, des-Arg9, [Leu8]-bradykinin. In the presence of 10 microM captopril, an angiotensin-converting enzyme (ACE) inhibitor, all coronary arteries examined in this experiment showed endothelium-dependent relaxation to fM bradykinin. These results show that some bovine coronary arteries relax in response to fM bradykinin, and this response is mediated predominantly by the release of nitric oxide via stimulation of endothelial B2-kinin receptors. The relaxation may be dependent on ACE activity. PMID- 9731954 TI - Pharmacokinetics of indomethacin in sheep after intravenous and intramuscular administration. AB - The pharmacokinetics of indomethacin (1 mg/kg) was determined in six adult sheep after intravenous (i.v.) and intramuscular (i.m.) injection. Plasma concentrations were maintained within the therapeutic range (0.3-3.0 microg/mL) from 5 to 50 min after i.v. and from 5 to 60-90 min after i.m. administration. After two trials, indomethacin best fitted an open two-compartment model. The mean (+/- SD) volumes of distribution at steady state (Vd(ss)) were 4.10 +/- 1.40 and 4.21 +/- 1.93 L/kg and the mean clearance values (ClB) were 0.17 +/- 0.06 and 0.22 +/- 0.12 L/h x kg for i.v. and i.m. routes, respectively. The elimination phase half-lives did not show any significant difference between routes of injection (t1/2beta = 17.4 +/- 4.6 and 21.25 +/- 4.44 h, i.v. and i.m. respectively). After i.m. administration, plasma maximum concentration (Cmax = 1.10 +/- 0.68 microg/mL) was reached 10 min after dosing; the absorption phase was fast (Kab = 26 +/- 18 h(-1)) and short (t1/2ab = 2.33 +/- 1.51 min) and the mean bioavailability was 91.0 +/- 32.8%, although there was considerable interanimal variation. In some individuals, bioavailability was higher than 100%. This fact combined with the slower elimination phase after i.m. than after i.v. administration, could be related with enterohepatic recycling. PMID- 9731955 TI - Pharmacokinetic studies of flunixin meglumine and phenylbutazone in plasma, exudate and transudate in sheep. AB - Flunixin meglumine (FM, 1.1 mg/kg) and phenylbutazone (PBZ, 4.4 mg/kg) were administered intravenously (i.v.) as a single dose to eight sheep prepared with subcutaneous (s.c.) tissue-cages in which an acute inflammatory reaction was stimulated with carrageenan. Pharmacokinetics of FM, PBZ and its active metabolite oxyphenbutazone (OPBZ) in plasma, exudate and transudate were investigated. Plasma kinetics showed that FM had an elimination half-life (t1/2beta) of 2.48 +/- 0.12 h and an area under the concentration - time curve (AUC) of 30.61 +/- 3.41 Lg/mL x h. Elimination of PBZ from plasma was slow (t1/2beta = 17.92 +/- 1.74 h, AUC = 968.04 +/- microg/mL x h). Both FM and PBZ distributed well into exudate and transudate although penetration was slow, indicated by maximal drug concentration (Cmax) for FM of 1.82 +/- 0.22 microg/mL at 5.50 +/- 0.73 h (exudate) and 1.58 +/- 0.30 microg/mL at 8.00 h (transudate), and Cmax for PBZ of 22.32 +/- 1.29 microg/mL at 9.50 +/- 0.73 h (exudate) and 22.07 +/- 1.57 microg/mL at 11.50 +/- 1.92 h (transudate), and a high mean tissue cage fluids:plasma AUClast ratio obtained in the FM and PBZ groups (80-98%). These values are higher than previous reports in horses and calves using the same or higher dose rates. Elimination of FM and PBZ from exudate and transudate was slower than from plasma. Consequently the drug concentrations in plasma were initially higher and subsequently lower than in exudate and transudate. PMID- 9731956 TI - The regulatory status of xylazine for use in food-producing animals in the United States. AB - Xylazine is commonly used in veterinary medicine as a tranquillizer or adjunct to surgical anaesthesia. Although its use is approved in companion animals and certain species of deer, xylazine remains unapproved for use in food-producing animals in the United States. This paper reviews existing toxicological and residue chemistry information on xylazine in food animals, particularly cattle, and discusses the regulatory status of the drug in the US, as well as the conclusions reached by the Joint FAO/WHO Expert Committee on Food Additives in its recent evaluation of xylazine. PMID- 9731957 TI - Quantification of flumequine enantiomers in plasma by high-performance liquid chromatography. PMID- 9731958 TI - Pathophysiology of scleroderma: an update. AB - OBJECTIVES: To review the pathophysiological background of systemic sclerosis in relation to the main components involved: microvascular system, immunological system and fibroblasts of the connective tissue. BACKGROUND: Although many particular aspects of the pathophysiology of systemic sclerosis have been investigated in recent years, the complexity of the pathogenesis and the important links between the components involved remain unclear. METHODS: Literature review. RESULTS AND CONCLUSION: Scleroderma is a connective tissue disorder resulting in a progressive fibrosis of skin and internal organs. The genetic background is not clear. The microvascular system is one of the first targets involved (damage of capillaries, enhanced expression of adhesion molecules interacting with lymphocytes, perivascular infiltrates as starting points for tissue fibrosis). The immune system is unbalanced (selection of T-cell subpopulations, elevated serum levels of several cytokines, occurrence of autoantigens to topoisomerase I, centromeric proteins and RNA polymerases). As far as autoimmunity is concerned the triggering autoantigen is still unknown. Development of connective tissue fibrosis is prominent (subpopulations of fibroblasts with disturbed regulation of collagen turnover by TGF-beta, CTGF and collagen receptors (alpha1beta1, alpha2beta1)). Investigation of pathophysiology of scleroderma is effected by monitoring the serum levels for soluble mediators, by cell culture studies of affected and non-affected fibroblasts and EC, by studying environmentally induced forms of scleroderma and by studies using animal models. PMID- 9731960 TI - The efficacy of a new topical treatment for psoriasis: Mirak. AB - BACKGROUND: A new natural product for the treatment of psoriasis has recently become available in many European countries: Mirak. The Mirak Home Care Packs consist of natural spring water, volcanic earth and vitamin E cream. Recently, the efficacy of Mirak has come into question. As this treatment is used by many psoriasis patients in Europe, it is important for dermatologists to be informed about the clinical effects of the therapy. AIM: To evaluate the efficacy and side effects of the Mirak Home Care Packs. METHODS: By means of a placebo-controlled left/right comparison, both clinical and histological parameters were evaluated during 6 weeks of treatment. RESULTS: The reduction in induration was significantly greater in the Mirak-treated lesions than in the lesions treated with a placebo. A reduction in desquamation was found in both treatments; the difference between the treatments was not statistically significant. A decrease in number of proliferative cells in the Mirak-treated lesions was seen, but the difference with placebo-treated lesions was not significant. The other investigated parameters did not change during treatment. No side effects were seen. CONCLUSIONS: The Mirak Home Care Pack induces a modest therapeutic effect compared to placebo treatment, without significant side effects. Treatment with the Mirak Home Care Packs alone will probably not be able to compete with the already existing treatments for psoriasis. PMID- 9731959 TI - Contact allergens in patients with leg ulcers. AB - BACKGROUND: Contact dermatitis can complicate the treatment of leg ulcers and is an acquired phenomenon resulting from the use of topical medications. OBJECTIVE: To show the incidence of contact dermatitis reactions to topical medications applied to leg ulcers and to evidence changing trends in such reactions through comparison of two case series about 20 years apart. SUBJECTS AND METHODS: We studied two groups of patients with leg ulcers that were patch tested with contact allergens in 1973-1974 and in 1994-1995. RESULTS: One or more positive patch tests was present in 75% and 40% of the patients, respectively. A decrease in the incidence of positive reactions to neomycin, local anesthetics and parabens mix was seen in 1994-1995. The most important contact allergens in 1994 1995 were fragrance mix, colophony and the excipients wool alcohols and amerchol. Other relevant sensitizers were formaldehyde, neomycin and gentamycin. CONCLUSION: The changing trends in contact allergens over the last 20 years may be explained by changes in the components of topical agents used for treatment. PMID- 9731961 TI - Clobetasol propionate followed by calcipotriol is superior to calcipotriol alone in topical treatment of psoriasis. AB - BACKGROUND: Although potent, topical corticosteroids offer effective and rapid healing of psoriatic lesions. Their long term use is limited because of the risk of side effects. Calcipotriol is safe for long-term treatment, but its initial efficacy is lower than with topical corticosteroids. OBJECTIVES: To investigate whether 2 weeks of treatment with clobetasol propionate 0.05% ointment bd followed by 4 weeks of treatment with calcipotriol 50 microg/g bd would offer therapeutic advantages over 6 weeks of continuous treatment with calcipotriol. METHODS: Forty-nine patients with moderate to severe plaque psoriasis were recruited from five centres in Norway. In a randomised, double-blind, right- versus left-side comparison, ointments were applied to two symmetrically-located areas. RESULTS: Two weeks of treatment with clobetasol propionate produced a significantly greater decrease in total symptom score (combined scores of erythema, induration and scaling) than calcipotriol treatment (P < 0.0001). This improvement on the clobetasol propionate-treated side of the body was maintained throughout a subsequent 4-week treatment period when calcipotriol was applied to both sides of the body (P < 0.0001). The superiority of the clobetasol propionate followed by calcipotriol treatment was maintained during a 4-week, treatment free, observation period. Treatments were well tolerated with no rebound effect. CONCLUSIONS: Clobetasol propionate ointment bd for 2 weeks followed by treatment with calcipotriol ointment bd for 4 weeks was superior to calcipotriol ointment alone in the treatment of plaque psoriasis. PMID- 9731962 TI - Aspergillus versicolor as cause of onychomycosis: report of 12 cases and susceptibility testing to antifungal drugs. AB - BACKGROUND: Onychomycoses caused by opportunistic moulds are not well understood, and many are due to Scopulariopsis brevicaulis and other species. Aspergillus versicolor is not documented as an etiological agent in most studies. We have found an increasing prevalence of this species which is involved in 5.8% of all fungal infections of toe nails. OBJECTIVE: To study the clinical and mycological characteristics of the onychomycosis caused by A. versicolor and the in vitro susceptibility of this mould to antifungal agents. RESULTS: Onychomycosis due to A. versicolor is mainly seen in people over 60 and presents with chronic involvement of the big toe nails. Predisposing factors are not always present and the infection does not respond to conventional topical antifungals. In vitro, A. versicolor has been shown to be resistant to griseofulvin and fluconazole as well as to amphotericin B, whereas MICs for itraconazole and ketoconazole are variable but within a range of 0.50-4.0 microg/ml; on the contrary, MICs for terbinafine are very low (<0.125 microg/ml). DISCUSSION: Aspergillus versicolor could be considered as an emergent pathogen causing toenail onychomycosis. Local treatment seems not to be effective. Of the various systemic antifungal agents studied terbinafine appears to be the most effective in treating onychomycosis. PMID- 9731964 TI - Squamous cell carcinoma of the skin--histopathological features and their significance for the clinical outcome. AB - BACKGROUND: Owing to the rising incidence of tumours, the question of reliable risk classification is becoming increasingly significant. OBJECTIVE: Participation in the multicentre carcinoma registry maintained by the Association of Operative and Oncological Dermatology of the German Dermatological Society requires, in addition to the parameters of clinical staging and grading already established by the International Union against Cancer, description of other histopathological criteria related to prognosis, with special attention to microstaging. METHODS: One hundred and eighty-four patients with squamous cell carcinoma of the skin were examined. The histological parameters, carcinoma type, Breslow index, invasion level, growth form, grading and mitotic index were recorded and classified, and clinical staging was performed. RESULTS: It was found that the clinical criterion of tumour diameter (T-category) determines the further course of the disease. The other parameters taken into account, namely pathohistological microstaging and grading, can increase the accuracy of prognosis evaluation, and in particular enable carcinomas of the T1-category to be classified as either high malignant or low malignant tumours. The desmoplastic squamous cell carcinoma subtype is a potential risk tumour. Endophytic tumours are more malignant in their development than exophytically-growing carcinomas, and the probability of recurrence and metastatic tumour spread further increases when ulceration can be detected by microscope. In order to distinguish metastatic and recurring carcinomas, in addition to determining the invasion levels after Clark, measuring the Breslow index proved to be an important criterion. Quoting the mitotic index augmented the grading system, which further improved the reliability of malignancy assessment. CONCLUSIONS: The TNM categories currently applied to squamous cell carcinomas of the skin must be supplemented by additional histological parameters which, according to our findings, permit more accurate classification of high and low malignant tumours. PMID- 9731963 TI - South American cutaneous leishmaniasis: report of ten cases in Israeli travelers. AB - BACKGROUND: Cutaneous South American leishmaniasis is caused by several species of leishmaniasis. Lack of appropriate treatment may lead to mucocutaneous leishmaniasis, mainly with L. b. braziliensis and L. b. panamensis. OBJECTIVE: To describe the clinical findings of Israeli travelers infected with cutaneous South American leishmaniasis and to draw attention to this problem. SUBJECTS: Ten patients were interviewed, examined and treated. RESULTS: Twenty-two lesions of cutaneous leishmaniasis were found, all in exposed areas. Patients were seen by an average three physicians (range 1-6) before the final diagnosis was confirmed by direct smear, after an average period of 125 days (range 88-270 days). Treatment with Pentostam was started after an average period of 134 days (range 94-275 days). All lesions healed completely, but with scarring. CONCLUSION: Travelers to endemic areas, as well as physicians, should be instructed about the potential risks and the clinical manifestations of cutaneous and mucocutaneous South American leishmaniasis. Such awareness will prevent undue delay in diagnosis and treatment. PMID- 9731965 TI - Hereditary multiple fibrofolliculomas, trichodiscomas and acrochordons: syndrome of Birt-Hogg-Dube. AB - The syndrome of Birt-Hogg-Dube (BHD) is an autosomal dominant syndrome, characterized by a triad of cutaneous lesions including multiple fibrofolliculomas, trichodiscomas, and acrochordons. There are many clinical expressions, solitary and multiple forms, with or without other skin tumors. In the literature BHD syndrome has been associated with internal malignancy. We describe a patient with multiple firm, skin-colored papules in which the three types of lesion are documented. No signs of systemic disease were present. PMID- 9731966 TI - Adult dermatomyositis with livedo reticularis and multiple skin ulcers. AB - Skin ulcers are commonly described in the juvenile form of dermatomyositis; however, in the adult form they are rarely described. We report on a patient with adult dermatomyositis, deep cutaneous ulcers in the skin folds and livedo reticularis eruption. PMID- 9731967 TI - Multinucleate cell angiohistiocytoma: a report of two cases. AB - We report the clinical, histological and immunological features of two cases of multinucleate cell angiohistiocytoma (MCAH) in women of 32 and 53 years of age, respectively. Clinically, MCAH occurs mostly in middle-aged women and consists of crops of reddish-purple, dome-shaped papules especially on the limbs. Histologically, the reticular dermis presents an increased number of small vascular channels with plump endothelial cells embedded in a fibrohistiocitic stroma with numerous bizarre multinucleate cells. Bizarre multinucleated cells are not specific to MCAH; they can be observed in numerous other cutaneous conditions. However, MCAH presents quite distinctive clinico-pathological findings and may be easily differentiated from other cutaneous disorders. PMID- 9731968 TI - Disseminated papular acantholytic dyskeratosis. AB - We present two cases of elderly women with clinically atypical transient disseminated papular eruption, showing suprabasal epidermal clefts with dyskeratosis and acantholysis. The clinical picture did not correspond to either Darier's disease, Hailey's disease or Grover's acantholysis. In spite of clinical similarity the cases differed in course and outcome. Overlapping dyskeratosis and acantholysis varied in different specimens. The case with transient acantholytic lesions of short duration showed much more pronounced dyskeratosis than the other in which several relapses occurred in a 4-year period. Such cases defying classification belong to the heterogeneous group of papular acantholytic dyskeratosis. PMID- 9731969 TI - Scleromyxoedema and thrombotic thrombocytopaenic purpura: two rare conditions both responding to plasma exchange. AB - We report the case of a 66-year-old female who over an 18-month period developed severe, disabling scleromyxoedema with pulmonary fibrosis. Treatment with oral prednisolone and melphalan had failed to prevent disease progression. Treatment with a 5-day course of plasma exchange, intravenous cyclophosphamide (500 mg) and methyl-prednisolone (1 g on 3 consecutive days) was unfortunately followed by the development of thrombotic thrombocytopaenic purpura (TTP). After 17 extra plasma exchanges, she recovered and there has been a dramatic improvement in her skin signs. We postulate that the extra plasma exchanges which she received as a consequence of developing TTP have contributed to this result. To our knowledge, TTP has never been associated with scleromyxoedema, but it is likely to be a coincidence in this case. PMID- 9731970 TI - Trichorhinophalangeal syndrome type I. AB - A 32-year-old short statured woman with alopecia, typical facies, shortened angulated fingers and toes with Trichorhinophalangeal syndrome type I (TRPS I) is reported. The absence of exostosis and mental retardation rule out TRPS II. The absence of generalized shortness of all phalanges, metacarpals and metatarsals distinguish it from TRPS III. Possibly the various types of Trichorhinophalangeal syndrome are genetically identical but have a varied clinical spectrum. PMID- 9731971 TI - Benign nodular calcification and calciphylaxis in a haemodialysed patient. AB - Several types of soft tissue calcification can be detected from radiographic evaluation of patients with end-stage renal failure. The factors that predispose to such calcification include an increase in CaxP product in serum, the degree of secondary hyperparathyroidism, the level of blood magnesium, the degree of alkalosis, and the presence of local tissue injury. Three major varieties include calcification of medium-sized arteries, periarticular or tumoral calcification and visceral calcification. Calciphylaxis is a phenomenon consisting of acute ischemic necrosis in presence of calcification of dermohypodermic arterioles. It occurs mostly in chronic renal failure patients with secondary or tertiary hyperparathyroidism with a persistently elevated calcium-phosphorus product. There are few options in treating calciphylaxis and the outcome is generally poor. The authors report the case of a haemodialised patient with benign nodular calcification and calciphylaxis. The coexistence of both entities in the same patient has never been described. PMID- 9731972 TI - Allergic contact dermatitis from naftifine in a child without cross-reaction to terbinafine. AB - Allergic contact dermatitis from naftifine has been scarcely described in the English literature, all of them in adults. We report a case of a 12-year-old girl who developed an acute eczema on her neck after application of a naftifine cream. This fact was confirmed by a patch-test study. We did not find a cross-reaction to terbinafine, a structurally linked allylamine. PMID- 9731973 TI - Cellulitis of the eyelids associated with sinusitis and brain abscess. AB - Erythema in the orbital area can indicate systemic and life-threatening diseases. We experienced an unusual and serious case of orbital cellulitis that was difficult to distinguish from a case with good prognosis. A 21-year-old man developed an erythema around his eyes. He exhibited no symptoms that would suggest lesions in deep tissues, but his condition turned out to be cellulitis retrogradely metastasized from an odontogenic sinusitis traced to a dental treatment problem. Computed tomography revealed complication of a large abscess in the frontal lobe. Cellulitis of the orbital area requires particular clinical discretion. PMID- 9731974 TI - Amelanotic lentigo maligna and amelanotic lentigo maligna melanoma: a report of three cases mimicking intraepidermal squamous carcinoma. AB - The clinical diagnosis of amelanotic melanoma may pose diagnostic difficulties. We report three cases of amelanotic lentigo maligna, two of which developed an invasive component (lentigo maligna melanoma). The clinical appearances in each case mimicked intraepidermal squamous carcinoma. PMID- 9731975 TI - Pityriasis rotunda. PMID- 9731976 TI - Amelanotic lentigo maligna. PMID- 9731977 TI - Chordoma. PMID- 9731978 TI - Kyrle's disease in diabetes mellitus and chronic renal failure. PMID- 9731979 TI - Pyodermia chancriformis: an unusual presentation of lymphomatoid papulosis. PMID- 9731980 TI - Acquired trichomegaly of the eyelashes in a child with human immunodeficiency virus infection. PMID- 9731981 TI - Ritodrine-induced pustular eruption in a pregnant woman with psoriasis. PMID- 9731982 TI - Coexistence of chronic discoid lupus erythematosus and sensorineural deafness. PMID- 9731984 TI - Heterogeneity of protein profiles of Helicobacter pylori isolated from individual patients. AB - BACKGROUND: In order to characterize the diversity of Helicobacter pylori (H. pylori) in infected individuals, 10 colonies of H. pylori were isolated from the gastric juice of 25 patients with gastroduodenal diseases (total 250 isolates). METHODS: Protein profiles of isolates were compared by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Results were confirmed by Western blotting (immunoblotting) test using rabbit antisera against three different strains of H. pylori. RESULTS: The protein profiles of 18 of 25 cases (72%) showed a single type of H. pylori with the same polypeptide pattern. In contrast, heterogeneity in the protein profiles was seen in isolates from seven cases (28%). Two differing H. pylori types with two very different polypeptide patterns were found in 10 isolates from one case. In six patients, the protein profiles of isolates were found to have variations in their polypeptides between molecular weights of 30,000 (30K) and 14K, which are thought to be associated with bacterial membrane protein. In some isolates, a polypeptide band of the 16K was missing. Each of three different antisera confirmed differences among the distinct isolates from individual patients. CONCLUSIONS: These findings suggest that more than one antigenically different strain of H. pylori may exist in same infected individuals. PMID- 9731983 TI - H. pylori and cagA: relationships with gastric cancer, duodenal ulcer, and reflux esophagitis and its complications. AB - BACKGROUND: Helicobacter pylori infection is accepted to be associated with two mutually exclusive diseases: duodenal ulcer and gastric cancer. Attention has recently focused on possible relationships between H. pylori and gastroesophageal reflux disease and its complications such as adenocarcinoma of the gastric cardia. The aim of this study was to provide a framework for explaining the seemingly paradoxical associations between H. pylori and various gastrointestinal diseases. METHODS: Available data regarding H. pylori infection, cagA, acid secretion, corpus gastritis, and gastroesophageal reflux disease (GERD) and its complications are reviewed, and testable hypotheses are presented. RESULTS: Linking the type of H. pylori (cagA-positive vs. cagA-negative), the pattern and intensity of inflammation, and acid secretion explains the apparent paradoxes in the associations between H. pylori and gastric cancer, duodenal ulcer, and GERD. Although H. pylori is inhibited by bile, a duodenal acid load sufficient to lower the average pH to precipitate bile acids overcomes that inhibition. H. pylori that contain a functional cag pathogenicity island produce a vigorous inflammatory response. The severity of mucosal inflammation predicts likelihood of different outcomes (e.g., in the bulb with likelihood of developing duodenal ulcer, and in the corpus with the degree of reduction in acid secretion and the rate of development of multifocal atrophic gastritis). Development of H. pylori corpus gastritis is promoted by profound inhibition of acid secretion (e.g., childhood infections or a high level of antisecretory therapy). The CagA protein, or the cagA gene, is a marker for enhanced inflammation, but CagA is not directly involved in the pathogenesis of gastric cancer or duodenal ulcer disease, nor is it a reliable indicator of the presence of a functional cag pathogenicity island. CONCLUSION: The relationship between the type of H. pylori infection, presence or absence of a functional cag pathogenicity island, corpus inflammation, and acid secretion explains the duodenal ulcer/gastric cancer paradox and the relationship between H. pylori infection and the complications of GERD. The predicted rank order for the presence of GERD and its complications (peptic stricture, Barrett's esophagus, and adenocarcinoma of the gastric cardia) is highest in the population without H. pylori infection, less in those with H. pylori infection, and least in those infected with cagA-positive H. pylori. Controversy and confusing epidemiological observations will continue unless future studies provide data on the gastric corpus histology (or acid secretion) as well as regarding the presence or absence of a functional and intact cag pathogenicity island of the infecting organism. PMID- 9731985 TI - Identification of potential diagnostic and vaccine candidates of Helicobacter pylori by "proteome" technologies. AB - BACKGROUND: There is great interest in characterizing the proteins of the gastric pathogen, Helicobacter pylori, especially those proteins to which humans respond immunologically. Such proteins have potential importance in diagnosis and vaccine development. METHODS: Two-dimensional gel electrophoresis in combination with Western blotting was used to separate and identify potential antigens of Helicobacter pylori strain Z-170. Proteins found to be reactive with pooled sera from 14 infected patients were individually digested in situ with endoproteinase Lys-C, and the resulting fragments were analyzed by matrix assisted laser desorption time-of-flight mass spectrometry (MALDI-TOF MS). RESULTS: Over 20 proteins were reactive in Western blots with pooled sera from 14 infected patients. The mass spectral data was compared with predictions from the H. pylori genome DNA sequence. Each of the 20 proteins was readily identified. CONCLUSIONS: We propose that this "proteome" approach for identification of previously unknown proteins will be useful in examining regulation of H. pylori gene expression and protein localization in the development of improved serologic tests to detect and monitor H. pylori infection. This approach will also be useful for identifying potential targets for antimicrobial or vaccine development for H. pylori and other pathogens whose genomes have been sequenced. PMID- 9731986 TI - Detection of coccoid Helicobacter pylori: light microscopical immunogold silver enhancing stain. AB - BACKGROUND: Helicobacter pylori (H. pylori) can be morphologically divided into spiral and coccoid forms. Although many different staining procedures for light microscopy have been developed to detect H. pylori, there are no reports that the coccoid forms can be specially identified by a staining method. The ability to identify the coccoid form of H. pylori by light microscopy would be important for studies evaluating its possible role in gastric disease. We examined whether both the coccoid and the spiral forms could be stained using anti-H. pylori antibody. METHODS: Specimens from the stomachs of dyspeptic patients with proven H. pylori infection and H. pylori from culture (1st and 4th day) were stained with the light microscopic immunogold silver enhancing stain using the rabbit IgG specific for H. pylori. Cultured H. pylori was also stained with electron microscopic immunogold staining method using the same antibody. The number of coccoid forms was counted by scanning electron microscopy. RESULTS: Biopsies and H. pylori from 6 patients were studied. H. pylori from both biopsy material and culture appeared as black organisms by light microscopic immunogold silver enhancing stain. The coccoid forms constituted 0.4% and 98.3% on days 1 and 4 of culture, respectively (by scanning electron microscopy). The antigen recognized by the anti-H. pylori antibody was located on the surface of the flagella, the cell wall, or in the cytoplasm by immunoelectron microscopy. CONCLUSION: This study shows that both spiral and coccoid forms of H. pylori can be detected by light microscopic immunogold silver enhancing stain using anti-H. pylori antibody. This simple stain is to be proven useful for studies aimed at understanding whether the coccoid form plays a role in disease. PMID- 9731987 TI - Effect of H. pylori infection and CagA status on leukocyte counts and liver function tests: extra-gastric manifestations of H. pylori infection. AB - BACKGROUND: It has been suggested that H. pylori infection is associated with abnormalities in total leukocyte count as well as the number of basophils and lymphocytes. In addition, CagA seropositivity has been associated with an increase in serum transaminase (SGOT) values. The aim of this study was to confirm the findings of previous subgroup analyses in patients before and after treatment for H. pylori infection and to ascertain whether the abnormalities reversed following successful treatment. METHODS: Blood counts and serum transaminase levels were obtained prior to and following treatment of H. pylori infection of H. pylori-infected duodenal ulcer patients. CagA status was assessed by Western blot of the H. pylori isolates obtained from the patients. RESULTS: Ninety-four ulcer patients were studied, including 77 with CagA-positive H. pylori isolates (82%) and 17 with CagA-negative H. pylori isolates. All study parameters remained within normal limits both before and after therapy. There were no significant changes in any study parameter in those who failed therapy. Successful therapy resulted in a significant fall in total white cell count (7413 +/- 520 cmm to 6738 +/- 410 cmm, for pretreatment vs. cured, respectively, p = 0.04) and was almost entirely accounted for by a reduction in the number of circulating polymorphonuclear leukocytes (4595 +/- 370 cmm to 3855 +/- 270 cmm for pretreatment vs. cured, respectively, p = 0.015). The pretreatment SGOT and basophil count were significantly higher in those with CagA-positive H. pylori (SGOT = 23 +/- 1 vs. 18.5 +/- 1 U). Successful or failed therapy with follow-up for 3 months post therapy did not result in a significant change of SGOT levels. CONCLUSIONS: We confirmed an increase in total leukocyte count and number of polymorphonuclear leukocytes in those with H. pylori infection. We also confirmed higher SGOT levels with CagA-positive H. pylori infection, but the failure to resolve within 3 months of cure of the infection makes it unlikely to be a direct result of the H. pylori infection. PMID- 9731988 TI - Performance of a rapid whole blood test for Helicobacter pylori in primary care: a German multicenter study. AB - BACKGROUND: Serological rapid whole-blood tests for the detection of H. pylori are presently being promoted for use in primary care. We conducted a multi-center study to investigate the diagnostic accuracy of the Boehringer Mannheim Helicobacter pylori test (BM test), which is identical with the Cortecs Helisal test. PATIENTS AND METHODS: A previous diagnosis of H. pylori, a history of peptic ulcer diseases, or proton-pump inhibitor, bismuth or antibiotic use during the preceding month were exclusion criteria. The BM test was performed prior to endoscopy by 7 primary care physicians, 5 practicing gastroenterologists, or a single physician in the university hospital outpatient service. During endoscopy, antral and corpus biopsies were obtained for histology and rapid urease testing (RUT). H. pylori positivity was defined by histology and/or RUT as reference methods. H. pylori IgG-ELISA was performed additionally. RESULTS: Of the 203 patients included, 151 were H. pylori-positive by reference methods (74.4%). The overall accuracy of the BM test was 77.3%. Eight BM tests were indeterminate, and in the other 195 patients the test performed as follows: sensitivity 80.3%, specificity 81.3%, positive predictive value 92.9%, negative predictive value 57.4%. Using IgG-ELISA as reference, the BM test performance was similar. It also did not differ substantially among the three groups of physicians involved. CONCLUSIONS: We found the performance of the BM test to be insufficiently accurate, as both over- and underdiagnosis of H. pylori infection were not infrequent. This test needs to be improved before its use in primary care can be recommended. PMID- 9731989 TI - Risk of gastrointestinal bleeding associated with Helicobacter pylori infection in patients with hemophilia or von Willebrand's syndrome. AB - BACKGROUND: Many episodes of bleeding in the upper gastrointestinal tract are caused by Helicobacter pylori infection. Because these episodes present a life threatening complication in patients with bleeding disorders, we prospectively investigated the prevalence of H. pylori infection in patients with hemophilia A or B and with the von Willebrand syndrome. METHODS: Seventy patients (54 men, 16 women, ages 40 +/- 11 years) and 100 age-related volunteers (63 men, 37 women, ages 39 +/- 9 years) were tested for H. pylori infection using the 13C urea breath test. Fifty-four patients with hemophilia and 16 patients with von Willebrand syndrome participated. RESULTS: Thirty-three (33%) of the controls and 24 (34.3%) of the patients showed positive 13C urea breath tests (p = .97). Nineteen (35.2%) patients with hemophilia and 5 (31.3%) patients with von Willebrand syndrome were positive for H. pylori. History of dyspeptic symptoms (28% vs. 26%) were not different in patients and controls (p = .91). Gastric ulcers (20% vs. 5%) and duodenal ulcers (7% vs. 5%) were diagnosed more often in patients with bleeding disorders. Fourteen of the patients (20%), but none of the controls had a history of gastrointestinal bleeding (p < .001). CONCLUSIONS: The rate of H. pylori infection and dyspepsia in patients with bleeding disorders is similar to the prevalence in the normal population. Due to increased bleeding complications, H. pylori screening and therapy appears mandatory in patients with bleeding disorders. PMID- 9731990 TI - Decreased prevalence of Helicobacter pylori infection in gastroesophageal reflux disease. AB - BACKGROUND: An increased incidence of reflux esophagitis has been reported after eradication of H. pylori in patients with duodenal ulcer. To determine if H. pylori is associated with lower rates of esophagitis, we studied the prevalence of H. pylori infection in patients with and without reflux esophagitis and a subgroup of patients with concomitant peptic ulcer disease. METHODS: Patients who underwent esophagogastroduodenoscopy and had diagnostic testing for H. pylori over a 30-month period were studied. H. pylori infection was determined by rapid urease testing, gastric histopathology, or serology. Reflux esophagitis was determined by endoscopic and/or histologic criteria. RESULTS: Of 514 patients, 39.5% had H. pylori infection and 22.2% had reflux esophagitis. The prevalence of H. pylori infection in patients with reflux esophagitis was 30.7%, compared with 42.0% in patients without esophagitis (p = 0.039). The odds ratio for esophagitis risk with H. pylori infection was 0.61 (95% CI, 0.39-0.95). Neither patient age nor gender affected H. pylori prevalence. In patients with duodenal ulcer, H. pylori was present in 36.4% of patients with esophagitis and in 69.2% of patients without esophagitis (p = 0.018). The odds ratio for esophagitis with H. pylori infection in these patients was 0.25 (95% CI, 0.09-0.73). CONCLUSIONS: Our study demonstrates that H. pylori infection is significantly less prevalent in patients with reflux esophagitis and may protect against its development. In duodenal ulcer patients, this effect was more dramatic. Further study is required to confirm these findings and elucidate mechanisms underlying possible beneficial effects of H. pylori. PMID- 9731992 TI - Lansoprazole quadruple therapy is effective in curing Helicobacter pylori infection. AB - BACKGROUND: Quadruple therapy using omeprazole combined with classic bismuth triple therapy has been advocated as optimal therapy for the cure of Helicobacter pylori (H. pylori) infection. We investigated the efficacy of substituting lansoprazole for omeprazole in proton pump quadruple therapy. MATERIALS AND METHODS: In a prospective open study, 219 consecutive patients, with either peptic ulcer disease or biopsy-proven H. pylori-associated gastritis, received seven days of lansoprazole, bismuth, tetracycline and metronidazole after three days of lansoprazole pretreatment. Cure of infection was judged by 14C urea breath test at six weeks after completion of therapy. RESULTS: On an intention to treat basis, 198 of the 219 patients (90%) were confirmed to be cured of H. pylori infection. Compliance was excellent and minimal side effects reported. CONCLUSION: Lansoprazole-based quadruple therapy achieves a very high cure rate in an unselected population of either peptic ulcer patients or those with H. pylori-associated gastritis. Recommended regimens should achieve at least 90% cure of infection. Lansoprazole quadruple therapy is effective and compares favorably with other H. pylori treatment regimens. PMID- 9731991 TI - Efficacy of lansoprazole in eradicating Helicobacter pylori: a meta-analysis. AB - BACKGROUND: The combination of lansoprazole with antibiotics either as double or triple therapy has demonstrated an H. pylori eradication rate of between 80 and 90%. With the aim of providing a complete revision of the results of these clinical studies and a quantification of the efficacy of lansoprazole in eradicating H. pylori and healing peptic ulcers, we have undertaken a meta analysis of all the controlled studies published in the literature. METHODS: This meta-analysis reviewed all randomized, controlled clinical trials published as full text articles in English between 1993 and 1996 that reported the efficacy of lansoprazole treatment as monotherapy or in combination with antibiotics in the treatment of peptic ulcer and in eradicating H. pylori. Articles were identified from the literature, which included both manual and computerized research (MEDLINE) and references provided by articles in this area. In order to compare the efficacy of triple therapy comprising lansoprazole vs. another PPI, data from abstracts (n = 5) were used, as no full text articles were located. RESULTS: This systematic review of the literature documents that lansoprazole has a high degree of efficacy in eradicating H. pylori, above all when used within treatment schemes including amoxicillin or clarithromycin, and metronidazole or tinidazole. This efficacy is comparable to that of other PPIs. CONCLUSIONS: Triple therapy allows the eradication of H. pylori in more than 85% of cases in patients with peptic ulcer. In addition, there is a substantial comparability of the efficacy of lansoprazole and omeprazole when they are used together with other anti infective agents. Thus, lansoprazole appears to offer an option in the eradication of H. pylori. PMID- 9731994 TI - Ranitidine bismuth citrate plus clarithromycin: a dual therapy regimen for patients with duodenal ulcer. AB - BACKGROUND: The combination of ranitidine bismuth citrate (RBC) and clarithromycin (CLR) was compared with each treatment alone for the eradication of H. pylori and healing of duodenal ulcers in patients infected with H. pylori. METHODS: This two-phase, randomized, double-blind, placebo-controlled, multicenter study evaluated 203 patients with an active duodenal ulcer treated with either (1) RBC 400 mg BID for 4 weeks plus CLR 500 mg TID for the first 2 weeks; (2) RBC 400 mg BID for 4 weeks plus placebo TID for the first 2 weeks; (3) placebo BID for 4 weeks plus CLR 500 mg TID for the first 2 weeks; or (4) placebo BID for 4 weeks plus placebo TID for the first 2 weeks. Patients with healed ulcers after treatment entered a 24-week observation phase for the assessment of H. pylori and ulcer relapse. RESULTS: Four-week ulcer healing rates were higher with RBC + CLR (71%) and RBC alone (66%) compared with placebo (15%; p < 0.05) and CLR alone (49%). H. pylori eradication rates were significantly higher with RBC + CLR (86%) compared with RBC alone (0%, p < .001) or CLR alone (24%, p < .001). Ulcer recurrence rates after 6 months were lower in patients eradicated of H. pylori infection (17%) compared with patients who remained infected (43%). All treatments were well tolerated. CONCLUSIONS: Ranitidine bismuth citrate plus clarithromycin is a simple, convenient, and well-tolerated dual therapy regimen that is effective in eradicating H. pylori and healing duodenal ulcers in patients infected with H. pylori. The eradication of H. pylori in patients with healed ulcers significantly reduces the rate of ulcer relapse. PMID- 9731993 TI - Cure of H. pylori infection using a 7-day triple therapy combining pantoprazole with two antibiotics. AB - BACKGROUND: Acid pump inhibitors combined with antimicrobials cure gastritis and peptic ulcer disease but a standard therapy has not yet been established. We therefore investigated a triple therapy with pantoprazole. METHODS: The aim of this open-label monocenter trial, involving 30 intention-to-treat patients with peptic ulcer disease or functional dyspepsia, was to assess the H. pylori cure rate after a 7-day triple therapy with pantoprazole (40 mg bid) plus metronidazole (500 mg bid) and amoxicillin (1 g bid). The H. pylori status was assessed by rapid urease test, histological examination and culture at the initial examination and by histological examination and culture at the study end 4 weeks after ending all therapy. RESULTS: At the end of the trial H. pylori was eradicated in 21 of 27 per protocol patients (78%; 95% CI 58-91%) and in 21 of 30 patients included in the trial (70%; 95% CI 51-85%). In 15 of 16 per protocol patients with metronidazole-sensitive strain (94%; 95% CI 70-100%) the infection was cured, but in contrast eradication was accomplished in only one of 3 patients with a metronidazole-resistant H. pylori strain. Post-treatment resistance to metronidazole was observed in 6 cases, although 4 of them had had H. pylori strains sensitive to metronidazole at the initial visit. The gastritis had clearly been improved, and the activity of gastritis had completely disappeared 4 weeks after treatment. Seven adverse events were observed in 7 patients, the intensity of which was moderate in 6 cases. CONCLUSIONS: This short-term triple therapy with pantoprazole, amoxicillin and metronidazole provides an effective regimen especially in patients with metronidazole-sensitive strain. PMID- 9731995 TI - Autoimmune reactions in type A and H. pylori gastritis. PMID- 9731996 TI - Male pattern hair loss: current understanding. PMID- 9731997 TI - Vulvodynia--a dermatovenereologist's perspective. PMID- 9731998 TI - Nehushtan: the dry bones, Ezekiel, Prometheus, the breath, and the signals. PMID- 9731999 TI - Double-labeled immunofluorescence study of cutaneous nerves in psoriasis. AB - BACKGROUND AND OBJECTIVE: In recent years, many reports have suggested an active role of neuropeptides in the pathogenesis of psoriasis. Increased numbers of neuropeptide-containing nerves positive for substance P (SP), vasoactive intestinal polypeptide (VIP), and calcium gene-related peptide (CGRP) have been reported in psoriatic tissue. As psoriatic epidermis has a larger mass/volume, however, it is expected to have more nerves and a higher number of neuropeptergic fibers. Therefore, instead of demonstrating a larger number of neuropeptergic fibers, a more significant study is to investigate whether the neuropeptergic fibers are denser in psoriatic tissue. In this study, we applied a double labeled immunofluorescence technique. This method allows the identification of the total number of nerve fibers and the number of nerves positive for specific neuropeptides. MATERIALS AND METHODS: We obtained biopsies from nine lesional and seven non-lesional psoriatic skins and six normal controls. Biopsies were snap frozen and then cut into 14 microm cryosections. The tissues were first treated with anti-microtubule associated protein (MAP)2 antibody to stain the nerves. This was followed by a second set of stainings for SP, VIP, and CGRP. Primary antibodies were used in dilutions of 1:200 for anti-MAP2, 1:200 for anti-SP, 1:800 for anti-VIP, and 1:400 for anti-CGRP. RESULTS: We found that the percentage of SP-positive fibers was twofold greater and the percentage of CGRP positive fibers was 2.5 times greater in the psoriatic epidermis than in the epidermis of normal skin. Psoriatic epidermis had 30.1 +/- 3.9% SP-positive nerve fibers compared with 15.7 +/- 3.7% in the normal control. The corresponding values for CGRP-positive nerve fibers were 30.1 +/- 3.9% and 12.0 +/- 4.2%. CONCLUSIONS: The results of our study suggest that SP- and CGRP-containing neuropeptide nerve fibers are more dense in the psoriatic epidermis. Both SP and CGRP are chemotactic to neutrophils and mitogenic to keratinocytes and endothelial cells. In addition, SP activates T lymphocytes and induces adhesion molecules on the endothelial cells. Our observations suggest that neuropeptides may play a significant role in the inflammatory and proliferative process of psoriasis. PMID- 9732000 TI - Oral lichen planus and HCV infection: a clinical evaluation of 263 cases. AB - BACKGROUND: Hepatitis C virus (HCV) infection induces variable dermatologic manifestations. Our purpose was to determine whether there is an association between HCV infection and oral lichen planus (OLP). METHODS: Antibodies to HCV were determined in patients with OLP (263 patients; 156 women and 107 men, with a mean age of 55.5 years) and in a control population. RESULTS: Seventy six cases (28.8%) were positive for HCV antibodies with the second-generation enzyme-linked immunosorbent assay (ELISA II) test. All of these cases were confirmed with the second-generation recombinant immunoblot assay (RIBA II) test. In 61 cases (23.1%), high levels of serum transaminase were found. Positivity for hepatitis B virus (HBV) markers was found in 31 patients (11.7%) and for hepatitis A virus (HAV) markers in 43 patients (16.3%). None had positivity for hepatitis D virus (HDV) markers. As a control group, we used 100 patients (58 women and 42 men, with a mean age of 55.3 years) referred to the School of Dentistry of the University of Naples "Federico II," and treated for general dental caries. In the control group, HCV antibody positivity was found in three cases. CONCLUSIONS: The high prevalence of HCV antibody in this group of patients with OLP, higher than in the healthy population, suggests a link (p = 1.423 x 10(-7), chi-squared test) between these two diseases. These findings stress the importance of liver examination in OLP patients, and the need for other studies on the high susceptibility to hepatitis viruses in the population in the southern part of Europe. PMID- 9732001 TI - Lupus erythematosus-like features in patients with cutaneous T-cell lymphoma. AB - BACKGROUND: The development of lupus erythematosus-like (LE-like) features in patients with cutaneous T-cell lymphoma (CTCL) has not been reported previously in the literature. Both diseases, however, have been etiologically linked to retroviruses. OBJECTIVE: Our purpose was to report four cases of patients with CTCL who developed LE-like features during the course of their disease, and to evaluate for evidence of antibodies to retroviruses in the sera of these patients. PATIENTS: Four patients with biopsy-proven CTCL with clinical or histologic features of systemic lupus erythematosus (SLE) were evaluated for clinical and laboratory criteria for SLE. Only one patient demonstrated four American Rheumatism Association (ARA) criteria sufficient for the diagnosis of SLE. The remaining three patients demonstrated one or two criteria for SLE. In addition, the sera of these patients were examined by Western blot analysis for evidence of human immunodeficiency virus type I (HIV-I), human T-cell lymphotrophic virus type I (HTLV-I), or human intracisternal A-type particle type I (HIAP-I) retroviral proteins. Each patient demonstrated antibodies to some of the retroviral proteins examined. The sera of two patients reacted to proteins for HIAP-I, and the sera of two patients reacted to p24 gag proteins of HIV-I. No patient reacted to HTLV-I proteins. CONCLUSIONS: Our report identifies four patients with CTCL who developed LE-like features during the course of their disease. Although the etiology of CTCL and SLE has not been well established, each has been linked to retroviruses. Evidence of antibodies to retroviral proteins was identified in each of our patients by Western blot analysis. Although the clinical and laboratory findings in these cases do not resolve the etiologic role of retroviruses in CTCL or SLE, they suggest that retroviruses may have a role in the pathogenesis of the clinical phenomenon reported in these four patients. PMID- 9732002 TI - The epidemiologic trends of scabies among Israeli soldiers: a 28-year follow-up. AB - BACKGROUND: Scabies is not a notifiable disease in most countries. Thus, the reported rates of the disease in large populations are usually inaccurate and based on estimations. Scabies is usually reported only when large outbreaks occur. OBJECTIVE: This article describes the global epidemiology of scabies in the Israel Defense Force. The data used in this study are based on the routine and mandatory reporting of every individual case of scabies to the Army Health Branch Epidemiology Department since 1968. RESULTS: There was a period of 13 years of quiescence from the implementation of reporting in 1968 until 1981. This period was followed by an epidemic of 15 years, peaking in 1985 and 1986, and returning to the baseline rates of the quiescent period by 1996. This prospectively observed pattern of at least 13 years of quiescence followed by a peak of 15 years is consistent with other, mostly retrospective, reports. Although the peak may have been triggered by the "Lebanon" war of 1982, it is noticeable that no peak was observed during the "Yom Kippur" war (1973). CONCLUSIONS: Prospective, mandatory reporting of the kind described here should help to better understand the epidemiology of scabies. PMID- 9732003 TI - Epidemiologic aspects of scabies in Mali, Malawi, and Cambodia. AB - BACKGROUND: The prevalence rates of scabies are compared in Bamako, Mali, Karonga District, Malawi, and Battambang Province, Cambodia. METHODS: In Mali, children attending three different urban schools catering for different socio-economic levels were examined specifically for scabies. In Malawi, data were collected during a total population survey for leprosy. In Cambodia, a sample survey was carried out in a rural area to determine the prevalence of leprosy and other skin diseases. RESULTS: In Mali, the prevalence rate of scabies among all the children examined was 4% (44/1103), but only 1.8% (7/388) in the higher socio-economic group. In Malawi, the overall prevalence rate of scabies was 0.7% (408/61,735). The highest rate (1.1%) was found among children 0-9 years of age. In Cambodia, the overall prevalence in the 13 villages screened was 4.3% (645/14,843). The highest rate (6.5%) was found among children 0-9 years of age. CONCLUSIONS: Scabies was most prevalent among children in Cambodia and Malawi, but there were considerable differences in the overall rates between the two areas studied. The data from all three countries indicate that poor socio-economic conditions, in particular crowding and public water supplies, are risk factors for scabies. PMID- 9732004 TI - Outbreak of rat mite dermatitis in medical students. AB - BACKGROUND: The tropical rat mite, Ornithonyssus bacoti (0. bacoti), is an ectoparasite on many rodents, but when the rodent is not available, humans may become the victim of the mite's bite. The bite induces a nonspecific dermatitis; therefore, it is not easy to diagnose rat mite dermatitis unless the parasites are found. MATERIALS AND METHODS: Ten cases of rat mite dermatitis were found in medical students who had studied in the same room of the library. Their nonspecific dermatitis consisted of small papules, and parasites were found in the skin or in the environment. The mites were collected and identified as O. bacoti, female. Histopathologic studies showed moderate perivascular lymphohistiocytic infiltrations intermingled with some eosinophils. The presence of rodents in or around the room was confirmed by the students, but there had been no preceding rodent eradication. Although the rats were not captured in the library, insecticides were sprayed, and no further problem with either mites or dermatitis developed during the follow-up period. CONCLUSIONS: Rat mite dermatitis can occur in clusters that involve a common source of exposure to the rat mite epidemiologically. Prompt identification of rat mites and the eradication of mites and rodents from the environment can prevent further spreading of the disease. PMID- 9732005 TI - A new model of epidermal culture for the surgical treatment of vitiligo. AB - BACKGROUND: Vitiligo can be successfully treated with grafts of autologous cultured epidermal cells. OBJECTIVE: To evaluate the efficacy of autologous grafting of epidermal cells, cultured by an original method, in the treatment of localized vitiligo refractory to other therapies. METHODS: Autologous normally pigmented skin was used to culture keratinocytes and melanocytes on a supporting layer of biomaterial (Laserskin), which was grafted directly onto achromatic skin after de-epithelialization with liquid carbon dioxide. The percentage area of repigmentation was calculated by image analysis. RESULTS: Initial repigmentation of the treated areas was observed 1 month after treatment. Repigmentation continued to increase for 3 months after grafting. Follow-up at 3, 6, 12, and 18 months showed almost complete repigmentation in six out of 11 cases. In four other patients, 40-71% of the grafted achromatic area was repigmented. In one patient, repigmentation was impeded by sepsis. CONCLUSIONS: The method was found to be effective in the treatment of localized vitiligo refractory to other treatments. The therapeutic procedure was simple, reproducible, and easy to use. PMID- 9732006 TI - The "gauntlet" of pellagra. AB - A 48-year-old alcoholic Filipino man presented to the outpatient department with a 2-year history of an eruption in a photosensitive distribution and episodes of mild diarrhea. He was otherwise in good health. Dermatologic examination revealed a browny-red coloration, with a sharply demarcated erythematous border, affecting both hands and lower forearms, where it was striking in its symmetry (Fig. 1). Around the neck, it was typical of a casal's necklace. The fronts and backs of the legs and the dorsa of the feet were also erythematous. The patient showed no evidence of mental confusion. A clinical diagnosis of pellagra was made based on the morphology, and treatment with nicotinamide 500 mg daily was instituted. The eruption quickly improved and resolved in 2 weeks (Fig. 2). PMID- 9732007 TI - Nevus depigmentosus and inflammatory linear epidermal nevus--an unusual combination with a note on histology. PMID- 9732008 TI - Wells' syndrome triggered by centipede bite. PMID- 9732009 TI - Scrofuloderma. PMID- 9732010 TI - Captopril-triggered linear IgA bullous dermatosis. PMID- 9732011 TI - Epidermolysis bullosa acquisita in a 6-year-old Japanese boy. PMID- 9732012 TI - Familial mandibuloacral dysplasia--a report of four cases. PMID- 9732013 TI - Dermatography as a new treatment for alopecia areata of the eyebrows. AB - BACKGROUND: Alopecia areata is considered to be an autoimmune disease. It consists of patchy hair loss of the scalp and the eyebrows, making it a disfiguring condition. This 10-year study was designed to assess the usefulness of the treatment of the eyebrows with dermatography as a relatively quick and simple method to obtain a cosmetically satisfactory result. MATERIALS AND METHODS: The eyebrow areas were covered with a halftone pattern of tiny dots of color pigments, using a Van der Velden Derma-injector, without anesthesia. On average, two to three dermatography sessions of 1 h were required. The follow-up was 4 years. RESULTS: Thirty three patients, most of whom had been previously treated with a sensitizer such as dinitrochlorobenzene (DNCB), were treated with dermatography. Four patients had also been treated by a beautician with a crude form of tattooing. The results in 30 patients were excellent. In three patients the results were good. CONCLUSIONS: Dermatography is a technique offering a good alternative for time-consuming, troublesome treatment modalities that often have considerable side-effects. With dermatography, no side-effects were found. PMID- 9732014 TI - Severe alopecia areata treated with systemic corticosteroids. AB - BACKGROUND: Treatment of severe alopecia areata is difficult, and most efforts to successfully treat this condition have been disappointing. Systemic corticosteroids have been demonstrated as an effective treatment of severe alopecia areata. METHODS: Eighteen patients with alopecia areata (extensive patchy and totalis universalis types) were treated with systemic corticosteroids. RESULTS: Satisfactory hair regrowth was achieved in seven patients (38.9%). Hair fall subsequently occurred in all of these patients on discontinuation or tapering of corticosteroid therapy. CONCLUSIONS: Systemic corticosteroid therapy does not prevent the spread or relapse of severe alopecia areata and, when complete regrowth is obtained, it is rarely maintained off therapy. PMID- 9732015 TI - Giovan Cosimo Bonomo (1663-1696): discoverer of the etiology of scabies. PMID- 9732017 TI - Treatment of "cutaneous self-injurious behavior" (CSIB) PMID- 9732018 TI - Thrombocytopenia associated with oral terbinafine. PMID- 9732019 TI - Demodecidosis manifested on the external genitalia. PMID- 9732020 TI - Pemphigus vulgaris associated with systemic lupus erythematosus. PMID- 9732021 TI - Cutaneous angiosarcoma arising on the radiation site of a congenital facial hemangioma. PMID- 9732022 TI - Cockroaches can vector human disease. PMID- 9732023 TI - A review of heartwater and the threat of introduction of Cowdria ruminantium and Amblyomma spp. ticks to the American mainland. AB - Heartwater, caused by the rickettsial agent Cowdria ruminantium, is one of the most devastating livestock diseases in sub-Saharan Africa. In addition to domestic cattle, sheep, and goats, a variety of nondomestic species can acquire subclinical and clinical infections. Recent epidemiologic findings that demonstrate a long-term host carrier state in domestic and wild ruminants, intrastadial transmission by the tick vectors (Amblyomma spp.), vertical transmission of the agent from cows to their calves, and the presence of both C. ruminantium and Amblyomma variegatum in the Caribbean suggest that the introduction of this exotic disease to the American mainland is a significant threat. Veterinarians working with wildlife should be familiar with this disease and should follow appropriate preventive measures to minimize the risk of infection in captive and wild populations of ruminants. PMID- 9732024 TI - Reproductive assessment of male elephants (Loxodonta africana and Elephas maximus) by ultrasonography. AB - Transrectal ultrasonography was performed on five wild and two captive male African elephants (Loxodonta africana) and four captive male Asian elephants (Elephas maximus) to develop standards for assessment of reproductive health and status. The entire internal urogenital tract was visualized ultrasonographically by using a 3.5 MHz or a 7.5-MHz transducer in combination with a probe extension adapted for elephant anatomy. The findings were verified by postmortem ex situ ultrasound examinations in several individuals of each species. Each part of the internal urogenital tract was sonographically detectable except for the bulbourethral glands and the cranial portion of the ureters and ductus deferentes, which were only observed in situ in the neonate. Each structure visualized was measured and described. The size and morphology of the urogenital structures, especially the accessory glands, were indicative of breeding status, if known. There was a notable difference between African and Asian males in the size and morphology of the prostate gland and a slight difference in the shape of the ampullae. No other structure showed significant species differences. The detection of the location and description of the testes may provide information for modifying present castration procedures. Furthermore, ultrasound examination of the male accessory glands may aid in the identification of potential semen donors for assisted reproduction programs in captive elephants. PMID- 9732025 TI - Evaluating adrenal activity in African wild dogs (Lycaon pictus) by fecal corticosteroid analysis. AB - A noninvasive corticosteroid hormone monitoring technique was validated for use in African wild dogs (Lycaon pictus). The double-antibody 125I radioimmunoassay for corticosterone was validated by demonstrating parallelism between serial dilutions of wild dog fecal extracts and the standard curve, recovery of corticosterone added to fecal extracts, and the time course of fecal corticoid excretion after an exogenous adrenocorticotropic hormone (ACTH) challenge. All feces were collected from three female and two male African wild dogs for 72 hr before and 144 hr after i.m. injection of long-acting ACTH (Acthar Gel, 400 IU). Fecal corticosterone immunoreactivity increased 10-30-fold within 24 hr of ACTH administration in all individuals, with peak concentrations from 1,200-8,000 ng/g. High-pressure liquid chromatography analysis revealed that >90% of all corticosterone immunoreactivity was associated with a single peak that exhibited intermediate polarity relative to cortisol and corticosterone reference tracers. Fecal corticosterone immunoreactivity appears to reflect adrenal activity in the African wild dog and, therefore, may be useful for evaluating stress. From a conservation perspective, these techniques can complement in situ and ex situ research studies designed to evaluate how environmental conditions and management strategies affect overall animal health. PMID- 9732026 TI - Health evaluation of free-ranging guanaco (Lama guanicoe). AB - Twenty free-ranging guanaco (Lama guanicoe) in Chubut Province, Argentina, were immobilized for health evaluations. All but two animals appeared to be in good condition. Hematology, serum chemistry, and vitamin and mineral levels were measured, and feces were evaluated for parasites. Serology tests included bluetongue, brucellosis, bovine respiratory syncitial virus, bovine viral diarrhea/mucosal disease, equine herpesvirus 1, infectious bovine rhinotracheitis, Johne's disease (Mycobacterium paratuberculosis), foot and mouth disease, leptospirosis (17 serovars), parainfluenza-3, and vesicular stomatitis. Blood samples from 20 domestic sheep (Ovis aries) maintained in the same reserve with the guanaco were also collected at the same time for serology tests. No guanaco had positive serologic tests. Sheep were found to have antibody titers to bovine respiratory syncytial virus, Johne's disease, leptospirosis, and parainfluenza-3. There was no apparent difference in external appearance or condition, or statistical difference in blood test values, between the animals that were positive or negative for parasite ova. PMID- 9732027 TI - Hematology and serum biochemistry values of free-ranging red howler monkeys (Alouatta seniculus) from French Guiana. AB - One hundred twenty-two wild red howler monkeys (Alouatta seniculus) were translocated during flooding of the forest at a hydroelectric dam site in French Guiana. Blood samples from 103 animals were evaluated for 13 hematologic and/or 22 serum chemistry parameters. Significant age-specific variation was found for white blood cell (WBC), lymphocyte, and platelet counts and for alkaline phosphatase values. Adult males and females had significant differences in red blood cell count, packed cell volume, and hemoglobin, creatinine, cholesterol, and calcium values. In juveniles, amylase and cholesterol levels were significantly lower in males than in females. Significant differences associated with reproductive status were also observed; i.e., lower cholesterol level in pregnant females and higher packed cell volume in lactating females. Chronic stress due to habitat disappearance may have been responsible for significant differences between thin animals and those in good condition. Thin animals had lower WBC, eosinophil, basophil, and monocyte counts and higher platelet counts. Capture stress was probably responsible for high and variable levels of creatine kinase, aspartate aminotransferase, and lactate dehydrogenase. PMID- 9732028 TI - A diet supplement for captive wild ruminants. AB - Nutritional husbandry of captive wild ruminants often requires feeding these animals a supplemental diet to enhance their health, reproductive performance, and productivity. Although supplemental diets for wild ruminants are commercially available, few have been evaluated in controlled intake and digestion trials. Voluntary intake, digestive efficiency, nitrogen retention, and gross energy utilization of pronghorn (Antilocapra americana), mule deer (Odocoileus hemionus), mountain sheep (Ovis canadensis), mountain goats (Oreamnos americanus), and wapiti (Cervus elaphus) consuming a high-energy, high-protein pelleted supplement were compared. Voluntary intake of dry matter, energy, and nitrogen were similar (P > 0.34) between mountain goats and mountain sheep and consistently lower (P < 0.03) for these species than for pronghorn, mule deer, and wapiti. Differences in digestive efficiency among species were inversely related to dry matter intake rates. Apparent digestibility of dry matter, organic matter, and neutral-detergent fiber was 10-20% higher for mountain goats and mountain sheep than for the other species (P < 0.04). Although these findings suggest a superior digestive efficiency for mountain goats and mountain sheep, species comparisons are inconclusive because of the confounding effects of season and ambient temperature on voluntary intake and digestion. Under the conditions of this experiment, the diet tested was safe, nutritious, and highly palatable. Protein and energy concentrations appear to be sufficient to meet or exceed known nutritional requirements of captive wild ruminants. PMID- 9732029 TI - (E)-5-(2'-bromovinyl)-2'-deoxyuridine inhibition of macropodid herpesvirus 1 in vitro. AB - Herpesviruses have caused the death of kangaroos and wallabies in European and North American zoos. Eight antiherpetic purine or pyrmidine nucleoside compounds were tested in plaque reduction neutralization tests for in vitro inhibition of macropodid herpesvirus 1, a virus that has been associated with illness in captive macropods in Australia. The virus was most susceptible to inhibition by (E)-5-(2'-bromovinyl)-2'-deoxyuridine (BVDU) and 5'-iodo-2'-deoxycytidine. Because BVDU effectively inhibits macropodid herpesvirus 2 in vitro, it may be the drug of choice for experimental therapy in herpesvirus infections in captive macropodids. PMID- 9732030 TI - A retrospective study of morbidity and mortality of raptors in Florida: 1988 1994. AB - A retrospective study was conducted on 390 raptors admitted to the University of Florida Veterinary Medical Teaching Hospital (VMTH) during 1988-1994. Representatives of 20 species were admitted; the five most common species were the barred owl (Strix varia, 72), eastern sreech owl (Otus asio, 63), red shouldered hawk (Buteo lineatus, 49), bald eagle (Haleaeetus leucocephalus, 43), and red-tailed hawk (Buteo jamaicensis, 38). A primary clinical diagnosis was determined in 340 (87%) of the 390 raptors admitted to the VMTH; a diagnosis was not made for the remaining 50 birds. Eighty-two percent (279) had traumatic injuries, and 87% (243) of those were directly related to human activity. The primary clinical diagnoses in the remaining 61 raptors included toxicosis (21), poor nutrition (15), infectious disease (11), orphaned young (11), and electrocution (3). The disposition of the 390 raptors was as follows: 61% (237) died or euthanized, 21% (80) released to the wild, 15% (57) outcome unknown, and 4% (16) permanent captives. Necropsies were performed on 32 of the 237 raptors that died. PMID- 9732031 TI - Long-term viral serology of semi-free-living and captive ungulates. AB - Between 1973 and 1994, blood samples were collected at Whipsnade Wild Animal Park (UK) from three ungulate species kept in enclosures, including 28 European bison (Bison bonasus), 37 scimitar-horned oryx (Oryx dammah), and 49 Pere David's deer (Elaphurus davidianus), and also from 144 semi-free-living Chinese water deer (Hydropotes inermis). These samples were tested for the presence of antibodies against three bovine viral diarrhea virus (BVDV)-like strains, three alpha herpesvirus strains, enzootic hemorrhagic disease virus (EHDV) of deer, bovine respiratory syncytial virus (BRSV), bovine adenovirus 3 (BAV-3), and enzootic bovine leucosis virus (EBLV). Thirty-three individuals (13.1%) had antibodies to one or more of the three BVDV-like viruses, with titers ranging from 1:5 to 1:16, and 17 individuals (6.6%) were positive for antibodies to one or more of the three alpha-herpesviruses, with titers between 1:4 and 1:80. The highest titers and greatest proportion of seropositivity were against SH9/11, a recently isolated cytopathogenic pestivirus from wild roe deer (Capreolus capreolus). There were no positive reactors to BRSV and EHDV, and there was only one BAV-3 positive reactor, a scimitar-horned oryx, and one EBLV reactor, a European bison. There is no serologic evidence that semi-free-ranging Chinese water deer are important reservoirs or transmitters of the viral diseases investigated. PMID- 9732032 TI - Bacterial isolates from California sea lions (Zalophus californianus), harbor seals (Phoca vitulina), and northern elephant seals (Mirounga angustirostris) admitted to a rehabilitation center along the central California coast, 1994 1995. AB - In 2 yr of bacteriologic culturing of 297 California sea lions (Zalophus californianus), 154 harbor seals (Phoca vitulina), and 89 northern elephant seals (Mirounga angustirostris) stranded alive along the California coast, the most frequent isolates from inflammatory lesions in live animals were Escherichia coli, Streptococcus viridans, Listeria ivanovii, beta-hemolytic Streptococcus spp., and Enterococcus spp. This is the first report of L. ivanovii isolation from a marine mammal. The common isolates from lung and liver in animals dying during rehabilitation were E. coli, Salmonella spp., Klebsiella spp., Pseudomonas spp., Aeromonas spp., Proteus spp., and Staphylococcus aureus. The most common isolates from brain were Enterococcus spp., E. coli, Klebsiella spp., and Pseudomonas spp. Ocular lesions were seen most often in harbor seals and elephant seals, from which the isolates cultured included Pseudomonas spp., Enterococcus spp., Streptococcus viridans, E. coli, Staphylococcus aureus, Proteus spp., Morganella morganii, Moraxella spp., beta-hemolytic Streptococcus spp., and L. ivanovii. Nine different Salmonella serotypes were isolated from 49 animals; S. newport was the most common. These results should enable those working clinically with these species to make logical decisions in choosing initial antimicrobial therapy. PMID- 9732033 TI - Alterations in blood platelet morphology during aggregate formation in the Asian elephant (Elephas maximus). AB - The ultrastructure of Asian elephant (Elephas maximus) platelets before and after activation with the agonist platelet activating factor (PAF) was studied. The unactivated platelet has a distinct ultrastructural appearance: the cytoplasm contains large randomly distributed granules but lacks the internal cristae that typify the open canalicular system in many types of mammalian platelets. Following PAF stimulation, large platelet aggregates form, but many platelets remain discrete, with little evidence of pseudopod formation or fusion of membranes. Two types of platelets are visible within the aggregates: those that are morphologically intact and those with gaplike features on the outer membrane and that have become degranulated, appearing as empty swollen sacs. The lack of platelet membrane fusion within the aggregates may permit the reversal of aggregation that is a characteristic response of elephant platelets to PAF. PMID- 9732034 TI - Gastrografin as a gastrointestinal contrast agent in the Greek tortoise (Testudo hermanni). AB - Eighteen Greek tortoises (Testudo hermanni), divided into three groups, were kept at three different average ambient temperatures. Gastrografin was administered to all individuals by orogastric tube at a dosage of 1 ml/130 g body weight. The partial and total transit times were recorded by means of radiographs taken immediately postadministration and at 20, 40, 60, 90, 120, and 150 min and 3, 4, 6, 8, 12, 24, 48, 72, and 96 hr postadministration. Mean total transit times were 2.6 hr (range 1.5-4.0 hr) at 30.6 degrees C, 6.6 hr (range 3.0-8.0 hr) at 21.5 degrees C, and 17.3 hr (range 8.0-24.0 hr) at 15.2 degrees C. These transit times allow a radiologic diagnosis within a relatively short period, especially compared with contrast studies performed with barium sulfate. The visualization of the intestinal tract is good with Gastrografin; however, intestinal mucosal detail was not completely satisfactory. PMID- 9732035 TI - Immobilization of wild kinkajous (Potos flavus) with medetomidine-ketamine and reversal by atipamezole. AB - As part of a wildlife rescue during the filling of a lake created by a hydroelectric dam (Petit Saut, French Guiana), 10 wild kinkajous (Potos flavus) were immobilized with medetomidine and ketamine for clinical examination and collection of biological samples. A mean (+/-SD) i.m. dose of 0.11+/-0.01 mg/kg medetomidine and 5.5+/-0.6 mg/kg ketamine rapidly induced complete immobilization (3.0+/-0.9 min) with good muscle relaxation and loss of corneal and pedal withdrawal reflexes. The duration and the quality of the anesthesia allowed procedures including minor surgery. Rectal temperature, heart and respiration rates, and relative oxyhemoglobin saturation (SpO2) were monitored at 5 min, 15 min, and 30 min after the medetomidine ketamine injection. Rectal temperature and heart rate significantly decreased during this time (P < 0.05). Low values of SpO2 (<90%) were recorded shortly after the injection. Hypoxemia partially resolved with time, confirmed by an increase in most SpO2 values. Atipamezole given i.m. at 5 mg/mg of medetomidine reversed the effects of the medetomidine in kinkajous. No adverse effects were observed during recovery. In group I, the antagonist was injected at 40.6+/-3.9 min. In group II, the animals showed signs of spontaneous recovery 37.9+/-6.9 min before antagonist injection at 52.2+/-6.1 min. Time from antagonist injection to ambulatory state was significantly shorter (P < 0.05) in group II (2.8+/-1.1 min) than in group I (6.9+/-1.2 min). PMID- 9732036 TI - Anesthesia in a Baird's tapir (Tapirus bairdii). AB - A Baird's tapir (Tapirus bairdii) was satisfactorily immobilized on two occasions with i.m. detomidine (0.065-0.13 mg/kg) and butorphanol (0.13-0.2 mg/kg). On the second occasion, anesthesia was induced by i.v. administration of ketamine (2.2 mg/kg). Twenty minutes later, endotracheal intubation was performed after an additional i.v. injection of ketamine (1.5 mg/kg). Anesthesia was maintained with isoflurane, which provided excellent conditions for radiology and surgery. Anesthesia was associated with hypoxemia when the tapir was allowed to breathe air and with hypoventilation. Mean arterial pressure remained satisfactory. No antagonist drugs were administered, and recovery from anesthesia was rapid and smooth. PMID- 9732037 TI - Heartworm (Dirofilaria immitis) disease and glomerulonephritis in a black-footed cat (Felis nigripes). AB - A 6-yr-old, 1.36-kg, intact female black-footed cat (Felis nigripes) was presented to the Veterinary Medical Teaching Hospital, University of Florida, with a history of depression, lethargy, and anorexia. Cardiac dysfunction and renal failure were diagnosed on the basis of antemortem and postmortem findings. At necropsy, heartworms (Dirofilaria immitis), glomerulonephritis, and endometritis were present. The glomerulonephritis could have been immune mediated and may have been associated with the heartworm infection or the chronic endometritis or both. Heartworm disease should be included in the list of differential diagnoses for any exotic cat housed outdoors in an endemic heartworm region that dies peracutely or has suggestive gastrointestinal or respiratory signs. Heartworm prophylaxis and annual serologic testing in exotic cats housed outdoors in heartworm endemic regions are recommended. PMID- 9732038 TI - Hepatotoxicity and secondary photosensitization in a red kangaroo (Megaleia rufus) due to ingestion of Lantana camara. AB - Three red kangaroos (Megaleira rufus), an adult male, an adult female, and a yearling, were exposed in bedding and food to coastal bermuda hay that contained the toxic plant Lantana camara. The adult male exhibited signs of anorexia, depression, lethargy, and jaundice. The adult female was presented dead. After 1 wk, following exposure to sunlight, the adult male and a yearling joey developed exudative dermatitis of the ear margins, eyelids, muzzle, and scrotum and opacity of the corneas. The adult male had a leucocytosis, anemia, bilirubinemia, bilirubinuria, hyperproteinemia, and elevated alanine aminotransferase, gamma glutamyl transpeptidase, alkaline phosphatase, and bile acid serum levels. Postmortem examination of the adult male revealed jaundice, and the liver was swollen, mottled, and pale yellow to reddish yellow. The gall bladder was markedly distended. Histopathologically, there was hepatocellular enlargement with vesiculation of the nuclei and sporadic feathery degeneration of the cytoplasm. The yearling joey survived and was treated symptomatically with i.v. fluids and antibiotics. The history, clinical signs, diagnostic findings, necropsy findings, and exposure to the toxic plant Lantana camara support the diagnosis of secondary photosensitization and hepatoxicity. PMID- 9732039 TI - Disseminated coccidioidomycosis in a mandrill baboon (Mandrillus sphinx): a case report. AB - A case of disseminated coccidioidomycosis caused by a dimorphic fungus Coccidioides immitis in a mandrill baboon (Mandrillus sphinx) was diagnosed following radiography, ultrasound-guided aspiration of thoracic lesions, and aspiration cytology of skeletal lesions of the left sixth rib. The diagnosis was confirmed by fungal culture and serum quantitative immunodiffusion for antibodies against C. immitis. PMID- 9732040 TI - Gastric spiral bacteria in small felids. AB - Nine small cats, including one bobcat (Felis rufus), one Pallas cat (F. manul), one Canada lynx (F. lynx canadensis), two fishing cats (F. viverrina), two margays (F. wiedii), and two sand cats (F. margarita), necropsied between June 1995 and March 1997 had large numbers of gastric spiral bacteria, whereas five large cats, including one African lion (Panthera leo), two snow leopards (P. uncia), one Siberian tiger (P. tigris altaica), and one jaguar (P. onca), necropsied during the same period had none. All of the spiral organisms from the nine small cats were histologically and ultrastructurally similar. Histologically, the spiral bacteria were 5-14 microm long with five to nine coils per organism and were located both extracellularly within gastric glands and surface mucus, and intracellularly in parietal cells. Spiral bacteria in gastric mucosal scrapings from the Canada lynx, one fishing cat, and the two sand cats were gram negative and had corkscrewlike to tumbling motility when viewed with phase contrast microscopy. The bacteria were 0.5-0.7 microm wide, with a periodicity of 0.65-1.1 microm in all cats. Bipolar sheathed flagella were occasionally observed, and no periplasmic fibrils were seen. The bacteria were extracellular in parietal cell canaliculi and intracellular within parietal cells. Culture of mucosal scrapings from the Canada lynx and sand cats was unsuccessful. Based on morphology, motility, and cellular tropism, the bacteria were probably Helicobacter-like organisms. Although the two margays had moderate lymphoplasmacytic gastritis, the other cats lacked or had only mild gastric lymphoid infiltrates, suggesting that these organisms are either commensals or opportunistic pathogens. PMID- 9732042 TI - Atrioventricular septal defects in three llamas (Lama glama). AB - Two half-sibling neonatal llamas (Lama glama) and one unrelated adult llama were presented with various complaints, including failure to thrive, respiratory distress, and excessive recumbency. The related camelids were born in successive years to the same dam but from unrelated sires. Thoracic auscultation revealed significant systolic and diastolic murmurs on both sides of the chest in all three llamas, and arterial blood gas evaluation revealed hypoxemia in two llamas. Echocardiographic examinations revealed large atrioventricular septal (AVS) defects in all three llamas. Two llamas were euthanized after diagnosis and the third died 4 mo later. Postmortem examination confirmed large AVS defects in all animals. There also was marked cardiomegaly in each animal. The discovery of such a cardiac anomaly in these three camelids suggests that it may be common in this species and may have a genetic basis. PMID- 9732041 TI - Cerebrospinal nematodiasis in a white-handed gibbon (Hylobates lar) due to Baylisascaris sp. AB - An adult white-handed gibbon (Hylobates lar) at a zoo in eastern Kansas was euthanized after developing a head tremor, generalized motor incoordination, and partial paresis of the right arm that persisted over 2 yr. Magnetic resonance imaging early in the course of the disease demonstrated a localized left frontal lobe cerebritis. Larvae morphologically consistent with a Baylisascaris species were seen in tissue sections of the cerebrum and cerebellum. Epizootiologic investigation, which included qualitative fecal flotations, evaluation of soil samples for nematode eggs, and necropsy examination of livetrapped raccoons (Procyon lotor), indicated that Baylisascaris procyonis was most likely to have caused the cerebrospinal nematodiasis in this gibbon. PMID- 9732043 TI - Actinomycotic splenitis and intestinal volvulus in an alpaca (Lama pacos). AB - Morphologic, microbiologic, and polymerase chain reaction amplification techniques were used to evaluate an alpaca (Lama pacos) with splenitis and intestinal volvulus. The intestinal volvulus produced a severe necrosuppurative typhlocolitis associated with vascular thrombosis and was most likely the cause of death of this animal. In addition, this animal had multiple coalescing abscesses affecting most of the splenic tissue. The isolation of Actinomyces spp. from the spleen and the morphology of the colonies when stained with Gram and Steiner stains support a diagnosis of splenic actinomycosis. PMID- 9732044 TI - Acral lick dermatitis in a jackal (Canis aureus). AB - Acral lick dermatitis was diagnosed in a 6-mo-old female jackal (Canis aureus) that was born and housed in a zoological garden in Hafez-Haim, Israel. Other dermatologic diseases were ruled out. Although the lesions were presumed to be psychogenic in origin, they resolved with topical therapy using an ointment containing benzocaine, neomycin sulfate, and hydrocortisone acetate. No recurrence has been observed. PMID- 9732045 TI - Antibody response to Newcastle disease vaccination in a flock of young houbara bustards (Chlamydotis undulata). AB - Twelve young houbara bustards (Chlamydotis undulata) were vaccinated with a lentogenic strain of Newcastle disease virus. Another seven birds were kept in close contact with the treated flock but were not vaccinated. Antibody levels were measured in all birds with hemagglutination inhibition test over the course of 1 yr. Antibody formation with no side effects was observed in 18 birds. The duration and amplitude of the antibody response differed between the groups. PMID- 9732046 TI - Prevalence of Salmonella and Campylobacter species in animals at Emperor Valley Zoo, Trinidad. AB - The prevalence of Salmonella and thermophilic Campylobacter species in animals kept at the Emperor Valley Zoo, Trinidad, was determined. Of the 433 animals from a total of 30 species sampled, 28 (6.5%) and 11 (2.5%) were positive for Salmonella and Campylobacter, respectively. The difference was statistically significant (P < or = 0.001: chi2). Overall, 12 stereotypes of Salmonella were isolated, with S. miami accounting for eight (25.8%) of 31 isolates. All Campylobacter isolates were C. jejuni, with nine (81.8%) of 11 isolates originating from birds. Reptiles had a high prevalence of Salmonella infection with a high probability for salmonellosis, but the risk of campylobacteriosis appears to be very low. PMID- 9732047 TI - Clinical challenge. Distal extremity necrosis. PMID- 9732048 TI - The role of cellular senescence in skin aging. AB - Higher organisms contain two types of cells: postmitotic cells, which never divide, and mitotic (or mitotically competent) cells, which can divide. Postmitotic cells include mature nerve, muscle, and fat cells, some of which persist for life. Mitotic cells include epithelial and stromal cells of organs such as the skin. Because postmitotic and mitotic cells differ in their proliferative capacity, they may age by different mechanisms. Normal somatic mitotically competent cells do not divide indefinitely. The process that limits the cell division number is termed cellular or replicative senescence. Replicative senescence is thought to be a powerful, albeit imperfect, tumor suppressive mechanism. It is also thought to contribute to organismic aging. Senescent cells undergo three phenotypic changes: they irreversibly arrest growth, they acquire resistance to apoptotic death, and they acquire altered differentiated functions. The growth arrest is very likely critical for the role of replicative senescence in tumor suppression, but may be less important for the aging of organs such as the skin. On the other hand, the altered differentiation may be critical for compromising the function and integrity of organs like the skin during aging. Senescent keratinocytes and fibroblasts appear to accumulate with age in human skin. Moreover, senescent cells express genes that have long range, pleiotropic effects - degradative enzymes, growth factors, and inflammatory cytokines. Thus, relatively few senescent cells might compromise skin function and integrity. Moreover, by altering the tissue microenvironment, senescent cells may also contribute to the rise in cancer that occurs with age. PMID- 9732049 TI - A relationship between thermotolerance and longevity in Caenorhabditis elegans. AB - Studies of aging in the nematode Caenorhabditis elegans have revealed a relationship between stress resistance and the rate of aging: Mutations which extend mean and maximum life-span also confer resistance to thermal stress. We review the molecular genetics of aging in C. elegans and introduce methods for obtaining novel mutants which display altered aging rates. We present the use of the "surrogate" phenotype of thermotolerance to develop a selection for novel mutations which slow aging. PMID- 9732050 TI - DNA repair and transcription in human premature aging disorders. AB - The human progeroid disorders Cockayne syndrome and Werner syndrome present with several clinical features that are associated with normal aging. These include distinct changes in the skin. The genes responsible for these conditions have recently been cloned and characterized. They both contain a characteristic helicase sequence, and helicase activity has been demonstrated using the purified Werner protein. Helicases are involved in a number of DNA metabolic transactions, including transcription, replication, and DNA repair. Cockayne cells are deficient in a special type of DNA repair, transcription coupled DNA repair, but they also appear to be defective in basal transcription. The diverse functions of the Cockayne protein are under intense study. Werner cells may have subtle defects in DNA repair, and possibly also in transcription. The biochemical clarification of the precise role of these gene products is likely to provide very significant clues into the mechanism of aging. PMID- 9732051 TI - Molecular mechanisms for the senescent cell cycle arrest. AB - Normal human diploid fibroblasts (HDF) have a finite proliferative life-span at the end of which they are arrested with a G1 phase DNA content regardless of the culture conditions. Serum stimulated senescent HDF fail to phosphorylate their retinoblastoma protein (pRb) and consequently do not express a large cohort of late G1 phase genes whose products are necessary for entry into S phase. Because pRb is believed to be phosphorylated sequentially in G1 phase by cyclin D-CDK4/6 and cyclin E-CDK2 complexes, we and others have investigated the status of these complexes in senescent HDF. There is little or no cyclin E-associated kinase activity in senescent IMR90 even though potentially active cyclin E-CDK2 complexes are present, suggesting the presence of an inhibitor. Likewise, cyclin D is complexed with its catalytic partners CDK4 and CDK6 in senescent HDF, but it is not known whether these complexes are active. p21Sdi1,Cip1,Waf1, a ubiquitous inhibitor of the activity of cyclin-CDK complexes, increases progressively throughout the life-span of HDF, but then declines again after the cells become senescent. In contrast, p16Ink4a, which binds monomeric CDK4 and CDK6 thereby preventing their binding to cyclin D, is increased dramatically at the time of senescence and remains high for at least 2 mo. Thus, it is possible that increased p21 initiates the senescent cell cycle arrest in normal cells, but p16 is important for the long-term maintenance of that arrest. PMID- 9732052 TI - Effect of aging on DNA repair and skin carcinogenesis: a minireview of population based studies. AB - In the general population, the risk of developing sunlight-induced skin cancer increases as age increases. During the aging process, many biologic factors contribute to the increased risk of developing cancer, including increasing cumulative carcinogenic exposure and increased cellular susceptibility to DNA damage induced by carcinogens. The latter is probably due to an age-related decrease in cellular DNA repair capacity (DRC). Secondary to this decrease in DNA damage repair associated with aging, oncogene activation and amplification also increase, as does the frequency of defects in tumor suppressor genes. All these factors lead to carcinogenesis. Population studies using peripheral blood lymphocytes, transformed lymphoblastoid cells, and primary skin fibroblasts have shown that human DRC decreases with increasing age. The decrease in DRC is also correlated with an increased mutation rate. Reduced DRC may thus be one of the underlying biologic mechanisms for the age-related increase in risk of skin cancer. Therefore, it is important to take the effect of age on DNA repair into account in population studies of DNA repair and skin cancer risk. PMID- 9732053 TI - Age-related changes in activation of mitogen-activated protein kinase cascades by oxidative stress. AB - Oxidative stress is thought to play a critical role in aging and the pathogenesis of human disease. Molecular studies of both the physiologic function of oxidants and the deleterious consequences of exposure to oxidative stress have suggested that signal transduction cascades may be targeted by oxidants. Here, we review recent studies from this laboratory examining the molecular basis for the activation of mitogen-activated protein kinases by oxidative stress and the influence of these pathways on cellular fate. We examine the association between constitutive activation of extracellular signal-regulated kinase (ERK) and cancer, and discuss how such mechanisms may contribute to oxidant-induced skin carcinogenesis. We also address the relationship between a decline in activation of this same pathway and the aged phenotype. In this regard, we review evidence that a decrease in activation of ERK by growth factor correlates with a reduced proliferative capacity in the isolated rat hepatocyte model, and we provide new data indicating that the activation of the ERK pathway in response to oxidant stimuli is also decreased with age. Further evidence demonstrates that this alteration is associated with both a reduced mitogenic response and a decline in hepatocyte cell survival in response to oxidative stress. Finally, we provide perspective on how modulations in ERK signaling may interplay with other changes in signal transduction cascades in the aging process. PMID- 9732054 TI - Apoptosis: a role in skin aging? AB - Prevailing theories view aging as an outcome of both programmed and stochastic events that occur over the lifetime of the individual. In this context, aging can be defined as a progressive decline in homeostasis and a period characterized by the inability of the organism to respond adaptively to stress. Apoptosis thus stands out as a potential key cellular process that may be affected during aging because apoptosis is both an important homeostatic mechanism and a protective cellular response to stress. In this paper we introduce the topic of apoptosis, its regulation, and its roles in epidermal homeostasis as a feature of normal keratinocyte differentiation and as a cellular endpoint of stress. To probe the question of whether apoptosis contributes to the process of skin aging, we present evidence for apoptotic dysregulation during aging in nonskin systems and discuss some findings suggesting that decreased efficiency of apoptosis may contribute to the alterations characteristic of intrinsic (chronologic) aging and extrinsic (photoaging) skin aging. PMID- 9732055 TI - Aging in epidermal melanocytes: cell cycle genes and melanins. AB - With aging, melanocytes become unevenly distributed in the epidermis. In light skin individuals, hypopigmentation is found in association with focal hyperpigmentation (lentigo senilis). Apparently this results from progressive loss of active melanocytes and focal increase in melanocyte proliferation and/or aggregation. This paper summarizes the present knowledge on aging of melanocytes in vivo and in vitro, with a focus on the role of melanin as a determinant for proliferation and terminal differentiation. We describe that excessive melanin deposition by cyclic AMP-inducing agents results in increased expression of the cyclin-dependent kinase inhibitors p27Kp-1 and p21SDI-1/Waf-1, increased binding of p16 to CDK4, and terminal differentiation. Importantly, the efficiency with which the melanocytes exit the cell cycle depends on the melanin background of the donor's cells. Melanocytes from skin types IV-VI that accumulate large amounts of brown black melanin (eumelanin), lose expression of the transcription factors E2F1 and E2F2, two key regulatory proteins, and withdraw from the cell cycle more rapidly than melanocytes from skin types I and II that accumulate red/yellow melanin (pheomelanin). Thus, we propose that terminal differentiation is a tumor suppressor mechanism that becomes less efficient under imperfect eumelanization. PMID- 9732056 TI - Molecular mechanisms of cutaneous aging: connective tissue alterations in the dermis. AB - Cutaneous aging is a complex biological phenomenon consisting of two distinct components, (a) the intrinsic, genetically determined degenerative aging processes and (b) extrinsic aging due to exposure to the environment, also known as "photoaging". These two processes are superimposed in the sun-exposed areas of skin, with profound effects on the biology of cellular and structural elements of the skin. This overview summarizes our current understanding of the mechanisms of innate versus extrinsic aging with emphasis on connective tissue alterations, primarily collagen and the elastic fiber network. We also introduce a novel transgenic mouse model, expressing a human elastin promoter-reporter gene construct, suitable for studies on biology and preventive pharmacology of the cutaneous aging. PMID- 9732057 TI - Photodamage: an historical perspective. PMID- 9732058 TI - Aging versus photoaging: postulated mechanisms and effectors. AB - The differences between intrinsic aging and photoaging are reviewed. The various model systems currently employed for the studies of aging and photoaging are discussed. Findings on age associated decrements in receptor/ligand mediated signaling as well as changes during cellular senescence in the expression of nuclear transcription factors are described. The role of telomere shortening and oxidative damage in the aging process is explained. At the cellular level, genetic and behavioral differences between aging and photoaging are illustrated with particular emphasis on changes in the structure and function of the tumor suppressor gene p53. PMID- 9732059 TI - Inhibition of UV-induced p53 mutations by sunscreens: implications for skin cancer prevention. AB - Ultraviolet (UV) radiation is a potent human carcinogen and it induces skin cancer in experimental animals. Recent studies have shown that unique mutations in the p53 tumor suppressor gene contribute to the development of human and mouse UV-induced skin cancers. Such mutations are also found in sun-damaged skin and actinic keratosis, suggesting that p53 mutations arise early during UV skin carcinogenesis. Our studies have shown that p53 mutations can be detected in UV irradiated mouse skin months before the gross appearance of skin tumors, suggesting that p53 mutations can serve as a surrogate early biologic endpoint in skin cancer prevention studies. Indeed, application of sun protection factor 15 sunscreens to mouse skin before each UV irradiation resulted in an 88-92% reduction in the number of p53 mutations. Because p53 mutations represent an early essential step in photocarcinogenesis, these results imply that inhibition of this event may protect against skin cancer development. PMID- 9732060 TI - Molecular mechanisms of intrinsic skin aging and retinoid-induced repair and reversal. AB - Past studies have shown that topical treatment of sun-exposed skin with all-trans retinoic acid improves the clinical and histologic appearance of the skin. This is associated with a reduction in matrix metalloproteinase elaboration and with expression of a newly synthesized collagenous matrix. Whether retinoid therapy might have a similar impact on the appearance of intrinsically aged skin is not known. This study, using human skin in organ culture and epidermal keratinocytes and fibroblasts in monolayer culture, show that retinoic acid stimulates growth of both keratinocytes and fibroblasts and stimulates extracellular matrix production by the fibroblasts. Adult skin from sun-exposed and sun-protected sites responds equally well to retinoic acid, whereas neonatal skin is much less responsive under the same conditions. The implications of this are (i) that retinoids may be able to repair intrinsically aged skin as well as photoaged skin, and (ii) that retinoids modulate human skin cell function in a manner that is age-related, and not simply a response to photodamage. PMID- 9732061 TI - Molecular mechanisms of photoaging and its prevention by retinoic acid: ultraviolet irradiation induces MAP kinase signal transduction cascades that induce Ap-1-regulated matrix metalloproteinases that degrade human skin in vivo. AB - Ultraviolet radiation from the sun damages human skin, resulting in an old and wrinkled appearance. A substantial amount of circumstantial evidence indicates that photoaging results in part from alterations in the composition, organization, and structure of the collagenous extracellular matrix in the dermis. This paper reviews the authors' investigations into the molecular mechanisms by which ultraviolet irradiation damages the dermal extracellular matrix and provides evidence for prevention of this damage by all-trans retinoic acid in human skin in vivo. Based on experimental evidence a working model is proposed whereby ultraviolet irradiation activates growth factor and cytokine receptors on keratinocytes and dermal cells, resulting in downstream signal transduction through activation of MAP kinase pathways. These signaling pathways converge in the nucleus of cells to induce c-Jun, which heterodimerizes with constitutively expressed c-Fos to form activated complexes of the transcription factor AP-1. In the dermis and epidermis, AP-1 induces expression of matrix metalloproteinases collagenase, 92 kDa gelatinase, and stromelysin, which degrade collagen and other proteins that comprise the dermal extracellular matrix. It is hypothesized that dermal breakdown is followed by repair that, like all wound repair, is imperfect. Imperfect repair yields a deficit in the structural integrity of the dermis, a solar scar. Dermal degradation followed by imperfect repair is repeated with each intermittent exposure to ultraviolet irradiation, leading to accumulation of solar scarring, and ultimately visible photoaging. All trans retinoic acid acts to inhibit induction of c-Jun protein by ultraviolet irradiation, thereby preventing increased matrix metalloproteinases and ensuing dermal damage. PMID- 9732062 TI - Increased intracellular macrophage inflammatory protein-1beta correlates with advanced HIV disease. AB - The objective of this study was to correlate between macrophage inflammatory protein-1beta (MIP1beta) and viral loads in untreated, HIV-infected individuals. For that purpose, HIV-positive patients were tested for number of copies of HIV RNA in plasma and for intracellular MIP1beta in freshly explanted CD8 and CD4 lymphocytes and monocytes. Results demonstrate that the levels of MIP1beta in the various cell populations were significantly higher in the HIV group than in age matched healthy individuals. Moreover, patients with low CD4 cell counts (<500/microl) and relatively high viral loads exhibited much higher levels of intracellular MIP1beta than patients with lower viral loads and CD4 counts >500/microl. We conclude therefore that although MIP1beta is induced in the various cell populations as a result of HIV infection in vivo, a high intracellular level of MIP1beta appears to be linked to a deterioration in the immune status of the patients. PMID- 9732063 TI - Rhesus macaques that become systemically infected with pathogenic SHIV 89.6-PD after intravenous, rectal, or vaginal inoculation and fail to make an antiviral antibody response rapidly develop AIDS. AB - A new simian-human immunodeficiency virus (SHIV) stock (SHIV 89.6-PD), derived from plasma of a rhesus macaque used for in vivo serial passage of virulence attenuated SHIV 89.6, produces systemic infection after intravenous, intravaginal, or intrarectal inoculation of rhesus macaques. Infection with this virus results in high levels of viral antigen in plasma, a precipitous decline in CD4+ T-cell counts, and a disease syndrome that is characteristic of AIDS. Rapid progression to disease was associated with failure to seroconvert to viral antigens, whereas longer survival was associated with production of antiviral antibodies. In intravenously inoculated animals, peak antigenemia occurred at 7 days postinjection (PI) and severe CD4+ depletion occurred at 14 days PI. In mucosally infected animals, peak antigenemia occurred at 14 days PI and severe CD4+ depletion was not evident until 21 days PI. The 1-week delay in both viral antigenemia and CD4+ T-cell decline in mucosally infected animals is consistent with the hypothesis that, following vaginal inoculation, virus dissemination proceeds in a stepwise manner from the mucosal surface to the draining lymph nodes and subsequently to the bloodstream. This animal model can be used to test the ability of HIV-1 envelope-based vaccines to prevent infection or disease after challenge by the three major routes of HIV transmission. PMID- 9732064 TI - Efficacy of adding indinavir to previous reverse transcriptase nucleoside analogues in relation to genotypic and phenotypic resistance development in advanced HIV-1-infected patients. AB - We assessed the efficacy of adding indinavir in patients with advanced HIV-1 infection, who were previously exposed to different reverse transcriptase (RT) nucleoside analogues. Twenty-five patients with an initial median CD4 cell count of 20 cells/mm3 (range, 0-80 cells/mm3) were treated with indinavir (800 mg three times per day) for 24 weeks. The median initial viral load was 5.4 log (range, 3.6-6.7 log). Of these patients, 56% (14 of 25) had an initial decrease in viral load of >1 log and sustained response of >0.5 log of HIV-1 RNA from baseline. Twelve of these 14 responder patients (85%) showed a sustained RNA response undetectable by NASBA assay, and no genotypic changes in protease were detected at week 24. In those with a temporary or absent response to indinavir, either resistant viruses or lack of compliance was observed. In compliant patients (15 of 16), relatively small increases in 50% inhibitory concentration (IC50) to indinavir and only two to three amino acid changes were sufficient to produce treatment failure. Phenotypic drug-resistant assays at 24 weeks revealed cross resistance to ritonavir in all the patient isolates and to saquinavir in one third of the isolates. We observed an initial and persistent response to the addition of indinavir in patients with advanced disease and prolonged antiretroviral treatment. Therapy failure, as defined by increases in viral RNA, was associated with either lack of compliance or the development of low level indinavir-resistant virus. Clinical studies need to be designed to determine to what extent these viruses may respond to other protease inhibitors. PMID- 9732065 TI - Anemia and survival in HIV infection. AB - Several clinical studies have suggested that anemia is an independent risk factor for dying in patients with HIV disease. We analyzed data from a large urban HIV clinical practice in Baltimore to assess the annual incidence of anemia, the risk of dying in patients who develop anemia, and the association between recombinant human erythropoietin use to treat anemia and subsequent survival. In 2348 patients observed between 1989 and 1996, 498 (21%) developed at least grade 1 anemia (hemoglobin <9.4 g/dl); 95 (4%) developed grade 4 anemia (hemoglobin <6.9 g/dl). Development of anemia was associated with decreased survival, independent of other prognostic factors. Use of erythropoietin was more likely in patients of nonminority race, those who did not inject drugs, those with a lower CD4 count or AIDS, and those being treated for cytomegalovirus disease (p < .05). Adjusting for these factors as well as severity of anemia, age, diagnosis of opportunistic disease, blood transfusion, and antiretroviral therapy in a time-dependent Cox proportional hazards analysis, erythropoietin use (n=91) was associated with a decreased hazard of dying (relative hazard [RH]=0.57; 95% confidence interval [CII, 0.40-0.81; p=.002). Although we cannot rule out a treatment selection bias, adjusting for available prognostic factors and factors potentially associated with a decision to use erythropoietin suggests that erythropoietin for treatment of anemia is associated with improved survival in HIV disease. PMID- 9732066 TI - Factors of weight loss in patients with HIV and chronic diarrhea. AB - Weight loss is significant in patients with HIV and chronic diarrhea. The aim of our study was to test for the links between weight loss, the level of food intake, and the severity of diarrhea and nutrient malabsorption. One hundred and sixteen patients with HIV and chronic diarrhea underwent a standardized gastrointestinal and nutritional evaluation, which included a questionnaire on diarrhea, a prospective estimation of food intake, a measurement of blood parameters and fecal lipid and nitrogen outputs, a stool examination for bacteria and parasites, and upper and lower digestive tract endoscopy. Diarrhea resulted from an infection by Cryptosporidia, Microsporida, or other pathogens in 22%, 20%, and 13% of the patients, respectively. Diarrhea appeared idiopathic in 45% of the patients. A significant negative correlation existed between the severity of weight loss and the levels of nutrient intake (p < .005), and a significant positive correlation between the severity of weight loss and stool frequency (p < .01). Multiple linear regression identified low caloric intake and high stool frequency as predictive of weight loss. No significant correlation was found between weight loss and the parameters of malabsorption, either by bivariate study or multiple regression. These results suggest that, in patients with HIV and chronic diarrhea, the degree of wasting is significantly related to the levels of dietary intake and the clinical severity of diarrhea, but not to the extent of nutrient malabsorption. PMID- 9732067 TI - Prevalence of hepatitis G virus RNA in the sera of patients with HIV infection. AB - OBJECTIVE: The routes of transmission of the hepatitis G virus (HGV) are similar to those responsible for infection with HIV. We sought to evaluate the prevalence of HGV RNA in the sera of HIV-infected patients. METHODS: The sera of 157 HIV infected patients were assayed by reverse transcriptase-polymerase chain reaction (RT-PCR) using established primers for HGV. Patients were divided into group 1 (positive circulating hepatitis B surface antigen [HBsAg]), group 2 (positive anti-hepatitis C virus [HCV] antibody) and group 3 (without markers for HBV or HCV). RESULTS: The overall prevalence of HGV RNA was 22%; prevalence was higher in group 1 (49%) than in groups 2 (16%) or 3 (7%). Patients with positive HGV RNA had laboratory values similar to HGV RNA-negative patients except for higher CD4 counts. Patients with an estimated risk duration of < or = 14 years were more likely to be HGV RNA-positive than patients at risk for >15 years. HGV RNA was found as frequently in patients with a homosexual lifestyle as in injection drug users (IDU). Multivariable analysis showed that the presence of HBsAg was the strongest factor associated with the presence of HGV RNA in serum. CONCLUSIONS: Patients with HIV and HBV coinfection are significantly more likely to be HGV RNA positive. Patients with a risk factor duration for >15 years were less likely to be HGV RNA-positive, pointing to a decrease in HGV RNA prevalence over time. This study supports the notion that homosexual lifestyle, in addition to injection drug usage and blood product transfusion, is a risk factor for HGV infection. PMID- 9732068 TI - Discrepancy between sex- and water-associated risk behaviors for cryptosporidiosis among HIV-infected patients in San Francisco. AB - OBJECTIVE: Potential transmission of cryptosporidiosis through drinking water supplies has been highly publicized; however, it is unknown whether this reporting has increased patient awareness or reduced other risk behaviors for exposure to this organism, such as high-risk sexual behavior. METHODS: Consecutive patients presenting for initial evaluation to the Gastroenterology AIDS Clinic completed a questionnaire that assessed knowledge about cryptosporidiosis, perceived risk of infectious diarrhea, drinking water sources, and high-risk sexual behavior. RESULTS: Fifty-one patients completed the questionnaire (82% male; 86% homosexual; mean age, 38 years; median CD4 count, 136 x 10(6) cells/L). Most respondents (31 of 44; 70%) believed they were at risk for infectious diarrhea. Awareness of cryptosporidiosis was high (31 of 45; 69%) as was avoidance of tap water (26 of 51; 51%) and exclusive or frequent use of bottled or boiled water (40 of 51; 78%). Respondents who used bottled water reported spending an average of $331.76 U.S. annually. However, high-risk sexual behavior remained common: 21 (41%) of the 51 subjects reported unprotected anal intercourse or oral-anal sexual contact. High-risk sexual behavior was prevalent even among subjects who drank exclusively boiled or bottled water. CONCLUSIONS: Awareness of risk for infectious diarrhea and cryptosporidiosis is high among patients infected with HIV in San Francisco. Patients perceive drinking water to be a substantial risk factor for infectious diarrhea and incur significant expense to avoid tap water. However, high-risk sexual behaviors remain prevalent in this population and should be the focus of future education efforts. PMID- 9732069 TI - Determination of the inhibitory effect on Toxoplasma growth in the serum of AIDS patients during acute therapy for toxoplasmic encephalitis. AB - In 26 AIDS patients treated for toxoplasmic encephalitis, the inhibitory effect on Toxoxplasma growth of sequential sera taken before and after initiation of therapy was determined using a culture-based immunoassay and compared with pyrimethamine blood levels. A marked inhibition of Toxoplasma growth was observed 1 day after initiation of therapy with pyrimethamine plus sulfadiazine and was maintained between 67% and 93% throughout the 15-day follow-up period. The degree of inhibition was not correlated to pyrimethamine blood levels and seemed highly potentiated when sulfadiazine rather than macrolides was given in combination with pyrimethamine. PMID- 9732070 TI - Predictive factors influencing peak viral load drop in response to nucleoside reverse transcriptase inhibitors in antiretroviral-naive HIV-1-infected patients. AB - Therapy with two nucleoside reverse transcriptase inhibitors (NRTI), the backbone of triple combinations, is still widely used in early stages of HIV-1 disease. However, factors influencing virologic response need to be further analyzed, to test the hypothesis that the reduction of plasma RNA viremia with NRTI may be greater in patients with higher baseline viral load (BVL) and to analyze the predictive factors of viral load drop below detection (200 HIV RNA copies/ml of plasma). Selected for the study were 169 HIV-1-infected antiretroviral therapy naive patients with CD4+ T-lymphocyte counts ranging from 6 to 1040/microl coming from three randomized studies comparing the efficacy of monotherapy (zidovudine [ZDV], 250 mg every 12 hours; N=40) versus two-drug therapy consisting of ZDV (250 mg every 12 hours) with dideoxycytidine (ddC, 0.75 mg every 8 hours) or didanosine (ddI, 200 mg every 12 hours; N=129). Viral load was measured at 1, 3, and 6 months by polymerase chain reaction (PCR). A linear regression model was used to analyze the relation between BVL and the peak reduction of plasma RNA viremia. The variables included in a logistic regression model to determine the likelihood of VLs dropping below detection levels were age, gender, risk group for HIV-1 infection, baseline CD4+ lymphocyte count, BVL, clinical status (AIDS versus non-AIDS), HIV-1 phenotype (syncytium-inducing [SI] versus non-syncytium inducing [NSI]) and type of treatment (monotherapy versus double therapy). The peak reduction of VL was related to baseline level following a linear model in both monotherapy and double-therapy regimens. In the subgroup of patients treated with two NRTIs, the regression line that fitted best with the data was log10 (peak reduction)=1.8-0.36 log10 (BVL) (F=23.5; p < .0001). This indicates that for every increase of 1 log10 of BVL, the peak reduction would be of 0.64 log10 greater. Forty-nine (29%) of the 169 patients dropped to <200 copies/ml. The likelihood of dropping below detection level was significantly greater in those receiving double therapy versus monotherapy (odds ratio [OR]=16.1; 95% confidence interval [CI], 2-128), in those with baseline CD4+ lymphocyte count >350/microl (OR=2.28; 95% CI, 1.1-4.9) and in those with BVL <10,000 HIV-1 RNA copies/ml (OR= 2.25; 95% CI, 1.1-6.1). None of the 13 patients with an SI phenotype at baseline dropped below detection levels. The reduction of VL in response to two NRTIs was greater in those patients with a higher level of BVL. In conclusion, peak reduction below detection in response to NRTI can be predicted and is associated with double therapy, with a baseline CD4+ cell count >350/microl and with a BVL <10,000 RNA copies/ml. PMID- 9732071 TI - Determinants of sexual risk-taking among young HIV-negative gay and bisexual men. AB - Data from a cohort of young HIV-negative gay and bisexual men were analyzed to identify determinants of sexual risk-taking at baseline. Gay/bisexual men aged between 18 and 30 completed a self-administered questionnaire including demographics, depression, social support, substance use, and consensual versus nonconsensual sex. Risk-takers were defined as those who had unprotected anal sex with casual male sex partners in the previous year; non-risk-takers were defined as those who reported consistent condom use during anal sex with all male partners in the previous year. Logistic regression was used to identify independent predictors of sexual risk-taking. Of 439 men studied, risk-takers had less education, a higher depression score, less social support, and were more likely to report nonconsensual sex and recreational drug use relative to non-risk takers. Independent predictors of sexual risk-taking were low education, nitrite use, low social support (adjusted odds ratio [AOR]=1.65; 95% CI, 1.04-2.59), and nonconsensual sex experienced as a youth or adult (AOR=1.85; 95% CI, 1.15-2.96). Young gay/bisexual men reporting nonconsensual sex, low social support, or nitrite use were significantly more likely to have recently had unprotected anal sex with casual partners. HIV prevention programs aimed at young gay/bisexual men should include sexual abuse counselling and foster community norms supporting safer sex practices. PMID- 9732072 TI - HIV infection in disadvantaged out-of-school youth: prevalence for U.S. Job Corps entrants, 1990 through 1996. AB - To describe HIV infection prevalence and prevalence trends for disadvantaged out of-school youth in the United States, we analyzed the HIV prevalence for and demographic characteristics of youth, aged 16 through 21 years, who entered the U.S. Job Corps from January 1990 through December 1996. Job Corps is a federally funded jobs training program for socially and economically disadvantaged out-of school youth. All 357,443 entrants residing at Job Corps centers during their training were tested for HIV infection; 822 (2.3 per 1000) were HIV-positive. HIV prevalence was higher for women than for men (2.8 per 1000 versus 2.0 per 1000; relative risk [RR]=1.4; 95% confidence interval [CI]=1.2-1.6). Among racial/ethnic groups, prevalence was highest for African Americans (3.8 per 1000). Prevalence was higher for African American women (4.9 per 1000) than for any other gender and racial/ethnic group. From 1990 through 1996, standardized HIV prevalence-stratified by age, race/ethnicity, home region, population of home metropolitan statistical area, and year of entry--declined for women and for men: for women, from 4.1 per 1000 in 1990 to 2.1 per 1000 in 1996 (p=.001); and for men, from 2.8 per 1000 in 1990 to 1.4 per 1000 in 1996 (p=.001). These data suggest that HIV prevalence for disadvantaged out-of-school youth declined from 1990 through 1996. However, considering their youth, prevalence was still high, particularly for women and African Americans, most notably African American women. These data support the need for ongoing HIV prevention programs targeting such youth. PMID- 9732073 TI - High frequency of the GWG (Pro Trp) envelope variant of HIV-1 in Southeast Brazil. AB - The HIV-1 variant that contains the GWG amino acid sequence in the crown of the principal neutralizing determinant (PND) has been detected in a few patients in Japan, France, and Brazil by direct sequencing. We describe for the first time the use of restriction fragment length polymorphism (RFLP) and limited DNA sequencing of the C2-V3 region of the HIV-1 envelope (env) gene to determine the prevalence of the variant in 75 HIV-1-infected Brazilian patients. Overall prevalence of the GWG sequence as indicated by RFLP was 57% (43 of 75). The prevalence in females (72%) was higher than that in males (32%) and newborns (40%). Two GFG sequences and 1 GLG sequence were also detected. This finding is relevant for the planning of vaccines and for studies of the epidemiology of HIV 1 in Brazil. PMID- 9732074 TI - A study of sexual behavior among rural residents of China. AB - OBJECTIVE: Although the recent spread of HIV/AIDS and other sexually transmitted diseases (STDs) in China has been associated with sexual activities, little information has been available about sexual behavior in rural areas with high HIV prevalence. Studies identifying high-risk sexual behaviors are needed to formulate effective prevention programs. METHODS: A cross-sectional study design was used to measure sexual activities using a two-stage cluster sampling method. A two-part anonymous questionnaire was used. Sensitive questions related to sexual behavior were administered using a tape recorder, earphones, and an answer sheet which did not include the text. RESULTS: In total, 1057 subjects were interviewed. Among 886 sexually active individuals, 7.8% had >1 sexual partner, 22.8% had premarital sex, 2.4% had anal intercourse, 4.1% had oral intercourse, and 2.3% had both anal and oral intercourse. Less than 2% reported past or current sexually transmitted diseases. Overall, 10.4% used condoms; only 11.2% for every sexual act. History of premarital and extramarital sex was higher in younger people. CONCLUSIONS: Sexual norms in rural China are changing rapidly and high-risk sexual behavior among young rural residents is increasing. Strategies to prevent HIV/AIDS should include education to promote delayed onset of sexual activity, safer sexual behavior, and condom use. PMID- 9732076 TI - Caveat emptor. PMID- 9732075 TI - Determinants of progression of HIV infection in a Greek hemophilia cohort followed for up to 16 years after seroconversion. AB - Our objectives are to describe the progression of HIV disease and to assess the influence of hemophilia-related variables, age at infection, and antibodies to cytomegalovirus infection (anti-CMV) in a Greek cohort of 158 HIV-1-positive hemophilic men, who received prospective follow-up for up to 16 years after infection. A total of 79 patients had died, representing a cumulative progression rate of 72.4% (95% confidence interval [CI], 56.6-83.3). A significant proportion of the mortality (30%) resulted from conditions not formally related to AIDS, with liver failure and cerebral hemorrhage predominant. At 16 years after seroconversion, 66 patients had developed clinical AIDS, a cumulative progression rate of 58.2% (95% CI, 47.1%-86.3%) whereas 15 years after infection 81.5% (95% CI, 74.2%-87.9%) of the patients had AIDS or a CD4 cell count <200 cells/mm3. Hemophilia-related variables or presence of anti-CMV were not significantly associated with disease progression. Age at infection was a strong prognostic factor for all three endpoints. Appropriate modeling showed a nonlinear age effect, with a steeper increase of relative hazard for patients >40 years of age at seroconversion. The age effect remained significant even after controlling for current CD4 cell count. Further investigation is required to elucidate the mechanisms of the age effect and the contribution of HCV coinfection on the disease progression. PMID- 9732077 TI - Translaminar facet screw placement: an anatomic study. AB - Anatomic measurements for screw path length, caudal and lateral angles, and superior and inferior lamina border thicknesses from L1 to L5 were measured for translaminar facet screw fixation in 30 dried lumbar spines. All measured values were fairly constant from L1 to L5. The mean values of the length of the screw path and lateral angle gradually increased from L1-2 to L5-S1 levels (41 to 54 mm, 39 degrees to 60 degrees, respectively). The caudal angle of screw placement relative to transverse plane gradually decreased from L1-2 to L5-S1 levels (60 degrees to 38 degrees). The superior border of the lamina was relatively thinner, with the mean thickness ranging from a minimum of 1.3 mm at L1 to a maximum of 2.0 mm at L5. The thickness of the inferior border of the lamina increased from L1 to L5 (from 6.7 mm to 7.8 mm). This study confirmed that a translaminar facet screw, 40 mm to 50 mm long at L1 to L5 levels, 60 mm long at L5-S1 level, and 4.5 mm in diameter, should be inserted through the lumbar facet joint at an angle of 40 degrees to 50 degrees laterally at L1 to L5 levels, 50 degrees to 55 degrees laterally at L5-S1 level, 45 degrees to 60 degrees caudally at L1 to L5 levels, and 35 degrees to 40 degrees at L5-S1 level caudally. PMID- 9732078 TI - Assessing the accuracy of partial weight-bearing instruction. AB - A force-monitoring platform was used to ascertain accuracy in following instructions concerning partial weight bearing. Three groups of healthy volunteers were instructed to bear 60 pounds of weight on a foot in one of three ways: on a bathroom scale, against a therapist's hand, or on a force-monitoring platform. The accuracy of each group, tested against platform measures, revealed significantly greater accuracy in the group instructed on the force-monitoring platform. PMID- 9732079 TI - Salvage of failed amputation about the hip in peripheral vascular disease by open wound care and nutritional support. AB - Five consecutive patients with wound and/or plastic surgical flap failure after hip disarticulation or amputation at the lesser trochanteric transfemoral level were treated with local tissue debridement, open wound management, culture specific antibiotic therapy, and nutritional supplementation. All of the patients underwent amputation about the hip as a result of ischemic necrosis of the lower extremity. Four of the five patients were able to achieve wound healing by second intention. The fifth patient died 2 months after the surgery. None of the patients required revision surgery. One patient underwent split-thickness skin grafting to minimize the need for continued wound care. Local wound management combined with nutritional support and culture-specific antibiotic therapy is an acceptable alternative to major amputation stump revision in patients with potentially high morbidity who fail to heal after amputation about the hip. PMID- 9732080 TI - Orthopedic residents' perceptions of the content and adequacy of their residency training. AB - The content and adequacy of orthopedic surgery residency training can be evaluated by several means. The Accreditation Council for Graduate Medical Education and the Residency Review Committee set standards with which residency programs must comply in order to be accredited. Residents' perceptions of the content and adequacy of their training is another means of evaluating orthopedic residency training. A questionnaire was sent to all graduating orthopedic residents in the United States, Canada, and Puerto Rico. The questionnaire provided program and individual resident demographics, as well as the residents' rating of specific areas of residency training on a 5-point scale (1=superior, 2=above average, 3=average, 4=below average, 5=inadequate). Completed surveys were received from 454 of the 698 graduating orthopedic surgery residents listed by the American Academy of Orthopaedic Surgeons; the response rate was therefore 65.0%. Our respondents were representative of the entire population in terms of geographic and sex distribution. Respondents rated their general orthopedic training at 1.9. The areas of training that had the best ratings included trauma/fracture (1.8), adult reconstruction (1.9), and pediatrics (1.9). The worst rating was reported for training in foot and ankle (2.7). Factors related to better ratings for general orthopedic training included male sex of residents, programs with more full-time faculty, programs with more hours of weekly teaching conferences, programs with one or more faculty present at all teaching conferences and programs in which residents first operate independently at or before postgraduate year 4. Sixty-six percent of all respondents were planning to hold a fellowship immediately after graduation. The most common fellowships taken included sports medicine (20.5% of all respondents), hand (12.1%), and spine (9.5%). Younger graduating residents, those from larger programs (more residents per year), and those from the Mideast (U.S.), and New England regions were most likely to enter a fellowship after graduation. PMID- 9732081 TI - Acute compartmental syndrome from hematogenous osteomyelitis of the ulna. AB - Compartmental syndrome of the forearm in children is usually caused by fractures, soft-tissue damage, burns, or arterial injury. This report presents the case of a child who had compartmental syndrome of the forearm resulting from acute hematogenous osteomyelitis of the ulna. PMID- 9732082 TI - Irreducible anterior shoulder dislocation with fracture of the greater tuberosity. AB - The combination of an anterior shoulder dislocation and an avulsion fracture of the greater tuberosity can usually be reduced by closed methods. This report describes a patient requiring open reduction and division of the subscapularis tendon before reduction of the glenohumeral dislocation could be achieved. PMID- 9732084 TI - Hyphenated history: Osgood-Schlatter disease. AB - The language of orthopedics is rather interesting in that it often credits the original describers or those who popularized a disease process by attaching their names to the disease process in question. These so-called "eponyms" have become quite commonplace in our literature and offer important orthopedic historical insight. Often throughout history, the simultaneous discovery of a disorder is described by two independent researchers, resulting in a hyphenated eponym. Such is the case in the observations made by two physicians, Robert Bayley Osgood and Carl Schlatter, concerning overuse injuries of the tibial tubercle in adolescents. This disorder subsequently became known as "Osgood-Schlatter disease, and aspects of its hyphenated history are the focus of this paper. PMID- 9732083 TI - Foot pump prophylaxis for deep venous thrombosis: the rate of effective usage in trauma patients. AB - Trauma patients are at risk for deep venous thrombosis (DVT) but often cannot receive systemic anticoagulation therapy. The major reason for failure of mechanical methods of DVT prophylaxis is ineffective usage. It has been postulated that foot pumps may have a better compliance rate than do other devices. One thousand observations were performed on trauma patients in both the intensive care unit (ICU) and on the surgical ward. Foot pumps were applied properly and functioning correctly 59% of the time. Patients in the ICU had significantly better compliance than did patients on the surgical ward. These rates are not better than published rates for other devices for DVT prophylaxis. PMID- 9732085 TI - Pelvic prolapse: diagnosing and treating cystoceles, rectoceles, and enteroceles. AB - The current generation of women is maintaining a healthier and more active lifestyle into an older age. Treatable conditions such as stress urinary incontinence and pelvic prolapse detract from this active lifestyle. In many cases, an improved quality of life can be maintained by treating pelvic prolapse conditions with relatively minor surgical procedures. Optimal treatment requires a knowledge of pelvic floor anatomy, an understanding of the various pelvic floor defects, and experience in selecting the appropriate procedure. The unequivocal diagnosis of pelvic prolapse conditions can only be made on physical examination. Each section of the vagina -- anterior, posterior, lateral, and apex -- must be inspected and evaluated separately to define the true nature and degree of prolapse. The examination should be performed with a moderate amount of urine in the bladder, and the patient must strain forcefully during the procedure. In some cases, this requires that the patient stand or sit upright during part of the examination to allow all areas of prolapse to become manifest. When the proper procedures are performed, excellent long-term results can be anticipated. The successful treatment of cystoceles requires an evaluation for both lateral and central defects, as inadequate treatment of either defect will lead to recurrences. The treatment of rectoceles is more controversial: Most clinicians would repair symptomatic rectoceles, but many choose not to treat asymptomatic rectoceles because there is little documented benefit to justify the risk of postoperative dyspareunia. Small asymptomatic enteroceles may be treated with a pessary; however, large symptomatic enteroceles usually require surgery. PMID- 9732086 TI - Managing clinical complexities of long-term contraception. AB - The 3 methods of long-term contraception (LTC) approved for use in the US are intrauterine devices (IUDs), levonorgestrel subdermal implants, and sustained release medroxyprogesterone acetate injections. Women who use reversible LTC are likely to receive care from clinicians who did not prescribe the method of contraception originally. Fortunately, most concerns arising from the use of reversible LTC can be managed by clinicians regardless of their specialty. Problems associated with an IUD can include a missing string, partial expulsion of the device, change in menstrual pattern, vaginal discharge, or infection. Headaches, weight gain, dermatologic problems, changes in hair growth, and irregularities in menses are among the problems that clinicians may confront while caring for a woman who has had subdermal hormone implants or has been taking depot injections. Besides the problems caused by or complicating specific methods of LTC, a woman may seek clinical care for a variety of general concerns. These include a desire to terminate LTC in order to become pregnant, and physical changes that are suspected to indicate pregnancy or contraceptive failure. In addition, general health conditions such as concurrent medications, hypertension, and endocrine disorders may need special consideration in a woman using LTC. There are few medical indications for discontinuing or changing LTC, even when intercurrent illnesses arise. PMID- 9732087 TI - Recurrent miscarriage: causes, evaluation, and treatment. AB - Recurrent miscarriage or fetal loss syndrome (also known as fetal wastage syndrome) is characterized by recurrent spontaneous abortion. There are many syndromes associated with recurrent fetal loss, including anatomic anomalies, endocrine/hormonal abnormalities, genetic/chromosomal abnormalities, and blood coagulation protein/platelet defects. Many of these syndromes are treatable, leading to normal term pregnancy, if the clinician is astute and vigorously pursues a thorough evaluation of why the patient has suffered unexplained, spontaneous miscarriages. There is no uniform agreement on how many spontaneous, unexplained miscarriages are needed to diagnose recurrent fetal loss; we generally pursue an evaluation for causation if a women has had 2 or more such events. In this article, we discuss the common reasons for recurrent fetal loss, plus diagnostic procedures to consider in pinpointing the problem, such as cytogenetic studies, blood coagulation protein/platelet tests, hysterosalpingography, sonography, and magnetic resonance imaging. We also describe management strategies that often lead to successful pregnancy outcome when the underlying problem is addressed. For example, in the case of thrombotic defects, a common cause of recurrent fetal loss, we report a 100% success rate in achieving a normal-term delivery among women who took low-dose (81 mg/day) aspirin preconception followed by postconception low-dose (5000 units q 12 h) heparin. PMID- 9732089 TI - Assessing coronary artery disease in women: how useful is coronary angiography? AB - The data are conflicting, but the suspicion is that there may be a gender bias against referring women for angiography in coronary disease evaluation. These cardiologists, however, review the studies and conclude that, even with the available noninvasive tests, coronary angiography continues to be the gold standard for assessing and monitoring heart disease -- even in women. The burden of coronary artery disease (CAD) in women is significant. In spite of increasing uses of noninvasive testing, coronary angiography remains the gold standard in the diagnosis and assessment of CAD. Since gender differences exist in the clinical presentation of CAD and in the sensitivity and specificity of noninvasive testing, coronary angiography remains an invaluable tool in providing diagnostic and prognostic information in women. Angiography is also appropriate when vasospastic disease is suspected. Although gender differences in the indication for coronary angiography are recognized, evidence as to whether there is a bias against women in the referral for cardiac catheterization after noninvasive testing or myocardial infarction is conflicting. The possibility that physicians underestimate the risk of disease in women cannot be ruled out. Therefore, proper training of physicians in the clinical assessment and prediction of the pretest risk for coronary disease in women cannot be overemphasized. In addition, physicians should be aware that normal coronary angiograms in women cannot always rule out the existence of myocardial ischemia, especially in conditions such as variant angina and syndrome X. Coronary angiography has also been invaluable in elucidating the benefits of lipid lowering therapy and estrogen use in women in the prevention of heart disease. Coronary angiography, therefore, remains an invaluable tool in the management of CAD in women. PMID- 9732088 TI - Dysphoric disorders in women: a case of perinatal depression. AB - During her third pregnancy, a woman becomes preoccupied with illness and death. When the dysphoria continues after her child is born, how would you evaluate and manage the problem? PMID- 9732090 TI - Recognizing and treating syphilis in pregnancy. AB - The number of primary and secondary syphilis cases in young women rose dramatically in the late 1980s and early 1990s, due to illicit drug use and the exchange of drugs for sex. Of infants born to mothers with primary or secondary syphilis, up to 50% will be premature, stillborn, or die in the neonatal period; further, most of these children are born with congenital disease that may not be apparent for years. While appropriate treatment of the pregnant female can prevent congenital syphilis, the major deterrent has been the inability to effectively identify these women and get them to undergo treatment. In determining a penicillin regimen, the clinician must consider the stage of maternal infection, the length of fetal exposure, and physiologic changes in pregnancy that can affect the pharmacokinetics of antibiotics. Treatment decisions may be further complicated in patients who are allergic to penicillin or infected with HIV. The pathogenesis of congenital syphilis is not completely understood, but placental invasion is the presumed major route. All women should be screened for syphilis with a nontreponemal test (eg, rapid plasma reagin [RPR] or venereal disease research laboratory [VDRL] test) in the first trimester. Those at high risk should be retested at 28 weeks and near delivery. Even with appropriate treatment of syphilis during pregnancy, fetal infection may still occur in up to 14% of cases. Treating syphilis during pregnancy can be difficult due to physiologic changes that can alter drug levels and the risk that drugs will induce uterine contractions or compromise the health of the fetus. While there are added risks and potential complications, treatment regimens parallel those in nonpregnant women. PMID- 9732091 TI - Liver problems in pregnancy: distinguishing normal from abnormal hepatic changes. AB - Abnormal liver tests occur in 1 of 10 pregnancies, though liver function is usually normal during pregnancy. The data suggest that liver metabolic capacity may be reduced in late pregnancy. Hepatic excretory function has been assessed in human pregnancy by both bromosulfophthalein (BSP) and bilirubin tolerance tests. The data suggest that the hepatic excretion of both compounds is impaired in the last half of normal human pregnancy. Thus, the clearance of compounds that are metabolized via the microsomal oxidizing pathway or secreted into bile may be impaired during pregnancy (especially late pregnancy). There is a 20% increase in total body water during pregnancy, and cardiac output increases 30% to 50%. The increment in cardiac output represents shunting of blood to the fetal-placental unit. Serum cholesterol and triglyceride levels begin to rise in the fourth month of pregnancy and peak at term. At term, pregnant women have a 25% to 50% rise in serum cholesterol levels to 265+/-8 mg/dL and a 150% increase in serum triglyceride levels to 180+/-13 mg/dL. Chemical analysis of tissue samples and histologic studies suggest that both cholesterol and triglycerides accumulate in the liver during normal pregnancy. The latter is thought to represent a storage pool of metabolic fuel to sustain the fetus during periods of starvation or inadequate nutrition. It is believed that both the enlarged gallbladder and supersaturation of bile with cholesterol contribute to gallstone formation in pregnant women. PMID- 9732092 TI - Virtual consult--aggressive angiomyxoma of the vulva: impact of GnRH agonists. PMID- 9732093 TI - Gynecology case challenge: vaginal bleeding in a woman taking an injectable contraceptive. PMID- 9732094 TI - Gynecology case challenge--persistent amenorrhea postpartum. AB - A 32-year-old woman does not resume menses even more than a year after the birth of her third child. How would you assess and treat this problem? PMID- 9732095 TI - Endometrial ablation versus hysterectomy: STOP-DUB. AB - Dysfunctional uterine bleeding (DUB) is a common clinical condition that frequently leads to hysterectomy. Endometrial ablation --a "minimally invasive" surgical technique that removes or destroys the endometrial lining of the uterus - is a conservative alternative to hysterectomy for DUB. While endometrial ablation has lower immediate costs and shorter recovery than hysterectomy, symptoms are not always resolved. Available data from studies with admittedly incomplete follow-up suggest that up to one quarter of patients treated with endometrial ablation require repeat ablation or subsequent hysterectomy to stop DUB. This suggests that the short-term advantages of endometrial ablation may be offset by possible longer-term disadvantages. The Surgical Treatments Outcomes Project for Dysfunctional Uterine Bleeding (STOP-DUB) is a randomized trial designed to compare endometrial ablation against hysterectomy. The primary outcomes address issues of importance to women, such as quality of life and resolution of symptoms that led to surgery. Other outcomes include subsequent surgery and cost-effectiveness of the procedures. The study's target enrollment is 800 women--400 in each treatment group -- from 20 clinical centers throughout the US. The women will be followed for 2 years after surgery. Part of the STOP DUB is a parallel observational study that involves women who do not choose surgery or who are not eligible for the randomized trial but could become eligible with time. It is anticipated that the result of this research will provide important information to women and their health care professionals as they consider the relative merits of surgical treatments for DUB. PMID- 9732096 TI - Liver problems in pregnancy: part 2--managing pre-existing and pregnancy-induced liver disease. AB - In distinguishing normal from abnormal hepatic changes, the author described the expected changes in liver tests that occur during complicated pregnancy. This article reviews the forms of pre-existing liver disease that may affect or be affected by pregnancy, as well as liver diseases that tend to arise during pregnancy. Among the pre-existing liver diseases are autoimmune chronic active hepatitis, which may be activated by pregnancy and tends to be associated with an increased risk of still and premature births. Worsening of chronic hepatitis B and C has occasionally been observed. While some women with cirrhosis can sustain a normal pregnancy without any worsening of hepatic function, others develop liver failure; plus, women with cirrhosis are less fertile and have higher rates of both stillbirths and premature infants. Other liver disorders that may or may not be affected by pregnancy include Dubin-Johnson syndrome, Gilbert syndrome, benign recurrent intrahepatic cholestasis, Wilson's disease, hepatic adenomas, and focal nodular hyperplasia. Among the hepatic disorders that occur during pregnancy in normally healthy women and then resolve after delivery is intrahepatic cholestasis of pregnancy (also known as pruritus gravidarum, recurrent intrahepatic cholestasis of pregnancy, and obstetric hepatosis). Others include acute fatty liver of pregnancy and HELLP syndrome (hemolysis, elevated liver enzymes, and low platelet count), which may be part of the spectrum of disorders associated with pre-eclampsia/eclampsia. Pregnancy may also trigger the dissemination of herpes infection to the liver. PMID- 9732097 TI - Ob-Gyn interactive case challenge--liver disease in the third trimester of pregnancy. PMID- 9732098 TI - STD case challenge--differential diagnosis of a genital dermatosis. AB - A 25-year-old white woman arrives in your outpatient clinic with a red, nonpruritic genital rash. What is the differential diagnosis and treatment? PMID- 9732099 TI - Perspective on women's health: editors' 1997-1998 year in review. AB - Heart disease, breast cancer, and hormone therapy were top clinical concerns in women's health in 1997. One of the major reports on heart disease confirmed that women are no different from men in terms of early infarct-related artery patency rates, reocclusion after thrombolytic therapy, and ventricular functional response to injury/reperfusion; nevertheless, women have 3 times the mortality of men in the first 30 days after an acute myocardial infarction. Research brought only modest gains in the understanding of breast cancer etiology in 1997, but engendered major debate on whether women younger than 50 years should have mammograms every 1 to 2 years. A National Institutes of Health consensus conference said no, but the National Cancer Institute's National Cancer Advisory Board said yes. Evidence of estrogen benefits and risks mounted: One report added to the data suggesting that estrogen may retard age-related memory loss, while another study reported that the risk of breast cancer significantly increased with long-term use of estrogens. The interest in selective estrogen receptor modulators (SERMs), also called "designer estrogens," grew. Efforts to develop pharmacologic treatment for obesity suffered a setback in 1997 when a team reported that 1 in 3 patients who used d-fenfluramine developed abnormal valvular thickening, with the most severe cases needing valve replacement. One of the most promising events in colorectal cancer, the third most common cancer in women, was the set of screening guidelines issued by the Agency for Health Care Policy and Research. The year ended with major ethical debates about multiple gestation and cloning. PMID- 9732100 TI - Pelvic prolapse: diagnosing and treating uterine and vaginal vault prolapse. AB - Uterine prolapse is often associated with a concomitant rectocele, cystocele, and/or an enterocele. Moderate degrees of prolapse are often associated with a feeling of pelvic heaviness or fullness or low back pain. The symptoms usually worsen with exertion and ease with bed rest. In severe prolapse, the cervix may descend outside the vaginal introitus, and patients may complain that a "mass" is protruding from the vagina. Bleeding from mucosal ulcerations or from the cervical os may occur due to rubbing of the prolapsed tissue against the patient's clothing. The commonly associated problems of cystoceles and rectoceles may lead the patient to complain of difficulty voiding, recurrent urinary infections, and/or "splinting" to defecate. Mild cases of uterine prolapse do not require therapy unless the patient is symptomatic; in most cases of second- or third-degree prolapse, however, patients may be quite uncomfortable and desire therapy. Nonsurgical options, such as a pessary, are usually tried first if the patient desires conservative therapy. Operative repair for uterine prolapse is usually approached vaginally if the uterus is small. An abdominal approach may be preferred if the uterus is large or if the woman has had multiple previous pelvic procedures or has extensive endometriosis or other processes that may obliterate the cul-de-sac. In either approach, the uterosacral and cardinal ligaments must be carefully ligated and tied together, and the cul-de-sac must be obliterated to reduce the risk of subsequent enterocele and to properly suspend the vaginal vault. PMID- 9732101 TI - Managing psychiatric medications in the breast-feeding woman. AB - Given the high risk of postpartum psychiatric problems, clinicians need to be prepared to appropriately manage the breast-feeding woman who needs psychotropics. These psychiatric researchers examine the issues and offer guidelines. Following childbirth, many women are at high risk for the onset or recurrence of psychiatric illness. Women who need psychopharmacologic treatment may wish to breast-feed their infants, but the data regarding the degree of drug passage to the infant and the subsequent effects of this exposure on infant growth and development are very limited, leaving clinicians with little guidance for responding in ways that protect the health and well-being of both mother and infant. In general, the less protein-bound, the more lipid-soluble, and the more weakly basic a drug is, the more likely it is to diffuse into breast milk. When a psychotropic medication is administered, the infant's clinical status and serum concentrations, including metabolite concentrations, should be closely monitored. Among the agents that have been the subject of at least limited studies in breast feeding women are tricyclic antidepressants, selective serotonin reuptake inhibitors, benzodiazepines, and the mood stabilizers lithium, carbamazepine, and divalproex. This article examines the factors that influence infant exposure to psychotropic medication through breast-feeding and includes clinical guidelines for managing the breast-feeding woman on psychotropics as well as protecting and caring for her infant. PMID- 9732102 TI - alpha-MeGlc and D-glucose transport by hepatopancreatic brush border membrane vesicles from prawn. AB - Sugar transport by prawn (Penaeus japonicus) hepatopancreatic epithelium has been studied. Brush-border membrane vesicles (hBBMV) were isolated, studies of osmotic reactivity were made indicating that these vesicles were closed and with low contamination from basolateral membranes. Incubation of hBBMV in the presence of Na+ resulted in rapid sugar uptake by the vesicles with an overshoot at 5 min, achieving the equilibrium value at 60 min. The absence of Na+ or the presence of phloridzin inhibited the overshoot. This uptake appears to be dependent on the membrane potential, since K+ efflux down its concentration gradient in the presence of valinomycin accelerated sugar influx and increased the overshoot when K+-loaded hBBMV were used. The kinetic study of Na+-dependent sugar uptake reveals that values of Km and Vmax were of the same order of magnitud as those described for other absorptive epithelia. PMID- 9732103 TI - Serum proteolytic activities and antiproteases in human colorectal carcinoma. AB - Some proteolytic enzymes, trypsin, cathepsin B, cathepsin D, collagenase, elastase and their inhibitors, API and AMG, in serum of patients with colorectal carcinoma have been evaluated. Twenty patients belonged to stage B of colorectal carcinoma, twenty two patients to stage D (Astler and Coller classification) and a control group of thirty healthy volunteers were evaluated. Except in cathepsin D, patients exhibit higher enzymatic activities than healthy subjects, and both groups have all the proteolytic activities assayed in serum. Patients with disseminated disease have increased cathepsin B and collagenase levels, with a decrease of trypsin activity, showing an increment in API and AMG in sera. However, only the API values were significantly higher in patients with metastases. The coexistence of proteolytic activities in human sera together with their inhibitors is considered as well as the origin of these, tumoral and/or reactive, increments. Cathepsin B levels are raised in colorectal neoplasms and contribute to the destruction of the extracellular matrix and the proliferation of tumoral cells. There is evidence that a relation between collagenase like activity and tumor invasiveness exists. Cathepsin B and collagenase increases agree with the tumoral mass. On the other hand, trypsin decrease in metastatic carcinoma is probably related to the increment of their inhibitors, API and AMG, acute phase reactant proteins. PMID- 9732104 TI - Validation of an EIA kit for determination of total thyroid hormones in rat serum. Effects of different anaesthetics. AB - Two enzyme immunoassays for the quantitative determination of total thyroxine (T4) and total triiodothyronine (T3) in human serum were validated to measure total T4 and T3 in rat serum. The specificity, sensitivity, detectability and reproducibility have been tested as well as the effects of different anaesthetics (pentobarbital and a mixture of ketamine and xylacine) on serum thyroid hormone levels. Hormones were quantified, by means of a previously validated technique, 18 hours after surgery for the placement of a stem for blood extraction in conscious and freely moving animals. Thyroid serum levels, especially T3, were slightly lower in xylacine plus ketamine treated animals than in those treated with pentobarbital. The administration of glutamic acid, stimulated the secretion of thyroid hormones, this effect appearing 30 minutes after its administration and it was independent of the anaesthetic used during the surgery for the cannula placement, although in pentobarbital treated rats, the serum T3 level increase induced by glutamate appears 60 minutes after the treatment. PMID- 9732105 TI - Maternal body weight gain and fetus development of rats fed a moderately altered olive oil. AB - The present study examines whether the consumption of a moderately altered olive oil influenced body weight gain and food efficiency ratio of pregnant rats as well as placental and fetal development. Olive oil used for frying 15 times undergoes a relatively slight alteration involving a statistically significant increase in polar content (9.0+/-0.1 mg/100 mg oil vs 2.0+/-0.1 mg/100 mg oil; p < 0.001). The methyl ester content also increased (5.1+/-0.8 mg/100 mg oil vs 1.8+/-0.5 mg/100 mg oil; p < 0.02), while the linoleic acid and oleic acid contents decreased significantly (6.2+/-0.6% oil vs 7.2+/-0.2% oil and 75.8+/ 0.6% vs 78.9+/-0.2%, respectively, both p < 0.05). Wistar rats were divided into four groups, two of which included pregnant rats (P1 and P2) and the other two, non-pregnant rats (NP1 and NP2). Groups NP1 and P1 received a diet containing 15% of fat as unused olive oil, while groups NP2 and P2 were fed a diet with a fat content of 15% as the olive oil used in 15 fryings. Pregnancy increased food intake, body weight, weight gain and food efficiency ratio (P1 vs NP1, and P2 vs NP1), while consumption of the used olive oil diet with respect to the unused oil diet did not alter food intake, body weight, weight gain and food efficiency ratio, placental weight, fetal weight and the number of fetuses in P2 rats with respect to P1 ones. These results suggest that in pregnant rats consumption of olive oil with a moderate level of alteration, as the only dietary fat source, exerts no detrimental effects on the mother weight gain or conceptus development. PMID- 9732106 TI - Fluid electrolyte changes during prolonged restriction of motor activity in rat. AB - Water and electrolyte changes in urine and plasma of rats during prolonged restriction of motor activity (hypokinesia), have been studied, on 90 male Wistar rats (375 to 396 g) during a 15 day period of prehypokinesia and during a 90 day period of hypokinesia (HK). All rats were divided equally into two groups: rats placed under ordinary vivarium conditions served as vivarium control rats (VCR) and rats subjected to HK served as hypokinetic rats (HKR). The hypokinetic effect was carried out by keeping the HKR group in small individual cages that restricted all their movements in all directions without hindering food and water intake. During the 15 days of the prehypokinetic period and during the 90 days of the hypokinetic period, fluid consumed and eliminated in urine, food intake, body weight, plasma sodium and potassium concentration and excretion thereof in urine, plasma osmolality, total protein plasma concentration, whole blood haemoglobin and haematocrit concentration were measured. In the HKR group water and food intakes decreased significantly (p < 0.05) when compared with the VCR group, whilst diuresis, excretion of sodium and potassium in urine, plasma sodium and potassium concentration, plasma osmolality, plasma protein concentration, whole blood haemoglobin and haematocrit increased significantly (p < or = 0.05) when compared with the VCR group. It was concluded that prolonged exposure to HK induces significant changes in water balance and in both urinary and plasma sodium and potassium in rat. PMID- 9732107 TI - Histochemical study of the vastus lateralis muscle fibre types of athletes. AB - In 16 women and 66 men, aged 14 to 36 years, 10 sedentary and 72 athletes, the histochemical characteristics of the vastus muscle fibres have been studied. Muscle biopsies were processed histochemically using the myofibrillar ATPase method, and were classified according to gender, sport activity and type of exercise. The average diameter of muscle fibres was larger in men than in women and in trained individuals than in sedentary ones. The largest percentage of Type I fibres was found in long distance runners; the smallest in those performing karate and triple jump. The largest percentage of Type IIA fibres was found in swimmers and the smallest in footballers. The largest percentages of Type IIB and IIC fibres were found in footballers. The largest average diameters of Type I and Type II fibres were found in swimmers and in long distance athletes respectively. Type I and Type II fibres were dominant in the aerobic group and the anaerobic one, respectively; the percentage of Type IIC fibres was smaller in the anaerobic group. PMID- 9732108 TI - Effects of a single melatonin treatment on brain regional serotonin metabolism in the Syrian hamster. PMID- 9732109 TI - Body composition and biochemical profile as affected by diet and renal transplantation among renal patients. PMID- 9732110 TI - Assessment of upwind dinghy sailing performance using a Virtual Reality Dinghy Sailing Simulator. AB - The ability of fourteen competitive helmsmen of different skill levels to sail a standard course towards the wind (upwind) was assessed using a virtual reality sailing simulator. The simulator consisted of a Laser dinghy deck which pivoted between two supports and was dynamically controlled by a computer driven pneumatic ram. Computer generated graphics realistically reproduced helming, sheeting, tacking and boat trim. After familiarisation with the simulator, subjects performed a standard 1 km upwind test and were ranked according to their completion time. The subjects were then asked to fill out a questionnaire to obtain an estimate of how effectively the simulator reproduced the conditions of actual sailing. Mean scores showed the sailors considered overall feel and simulation of physical movement as "good" (5 on a scale of 1 to 6). Rankings for the upwind test were compared with independent competition rankings for each subject. Overall time to complete the upwind test correlated well with a subject's external ranking (Spearman's rank order r=0.99). The results indicate that the test used can differentiate between variations in upwind sailing performance over a wide range of ability. The simulator thus provides for the first time a method of measuring and analysing a sailor's performance in a controlled laboratory setting. PMID- 9732111 TI - Whole body cooling by immersion in water at moderate temperatures. AB - This study investigated the potential use of whole body cooling by water immersion for lowering body temperatures prior to endurance exercise. Rectal temperature (Tre), mean skin temperature (Tsk), oxygen consumption (VO2), and ventilation (VE) were measured in 7 male and 3 female subjects who were immersed in a water bath for up to 60 min. Initial water temperature was 28.8+/-1.5 degrees C and decreased to 23.8+/-1.1 degrees C by the end of immersion. Pre immersion Tre of 37.34+/-0.36 degrees C was not altered by 60 min water immersion but decreased to 36.64+/-0.34 degrees C at 3 min post immersion (p < 0.01). Tsk decreased from 33.23+/-1.4 degrees C to 26.95+/-1.8 degrees C (p < 0.01) at the end of immersion. Reductions in Tre and Tsk resulted in reduced body heat content (Hc) of approximately 545 kJ (p < 0.01) at the end of immersion. VO2 and VE increased from pre-immersion values of 0.34+/-0.08 L x min(-1) and 6.2+/-1.4 L x min(-1) to 0.54+/-0.09 L x min(-) and 11.5+/-5.4 L x min(-1) at the end of immersion, respectively. Heart rate remained unchanged throughout immersion. These results indicate that whole body immersion in moderately cold water temperatures is an effective cooling maneuver for lowering body temperatures and body Hc in the absence of severe physiological responses generally associated with sudden cold stress. PMID- 9732112 TI - Comparison of injuries in elite senior and junior Australian football. AB - Three thousand and thirty one AFL and 1034 injuries in the VSFL U/18 competition were recorded by club doctors over the 1992, 1993 and 1994 seasons. Hamstring strains had the highest incidence (86.4 per 10,000 player hours) and prevalence (30.2 hours missed per 1000 hours) of any injury in the AFL, but were significantly less common in the U/18 competition. Other injuries which were common in both competitions were ankle sprains, thigh haematomas, concussion, groin strains and head lacerations. Injury prevalence was higher overall in the AFL, with lower limb muscle strains (hamstring, calf, quadriceps) being significantly more prevalent than in the U/18 competition. Injuries which were significantly more prevalent in the U/18 competition included stress fractures and concussion. Subsequent to this study, coaches and medical staff in the U/18 competition were made aware of the high risk of stress fractures in young footballers with heavy training loads. The AFL injury survey is ongoing and in the process of being computerised; risk factors for specific injuries with high rates are being studied further. PMID- 9732113 TI - Measurement of elastic-like behaviour in the power squat. AB - Because traditional procedures of evaluating elastic-like behaviour have yielded mixed results, the purpose of this work was to explore two methods of measuring elastic-like behavior in the power squat. The entire concentric time method was based on traditional procedures. The initial concentric time method was developed to examine elastic-like behavior for the beginning 0.2 s of concentric movement. The present study compares a power squat performed maximally by nine subjects at 70% of their 1 repetition maximum. Squats were performed with rebound (REB) and without rebound (NRB). For the entire concentric time method only concentric time was significantly greater (p < 0.05) in the NRB than the REB. For the initial concentric time method the relative displacement, velocity, net work, & peak power of the center of mass were significantly greater (p < 0.05) in the REB than the NRB. Some subjects had theoretically infeasible negative results for elastic enhancement in the entire concentric time method, but not the initial concentric time method. It seems that measuring elastic-like behavior near the end of the movement can be confounded by the constraints of the task. Based on its success, the initial concentric time method appears to be more appropriate for measurement of elastic-like behavior in lifting. PMID- 9732114 TI - The relationship between repeated sprint ability and the aerobic and anaerobic energy systems. AB - A large number of team games require participants to repeatedly produce maximal or near maximal sprints of short duration with brief recovery periods. The purpose of the present study was to determine the relationship between a repeated sprint ability (RSA) test that is specific to the energy demands of Australian Rules football (ARF), and the aerobic and anaerobic energy systems. Seventeen ARF players participated in the study. Each participant was assessed for VO2 max, accumulated oxygen deficit (AOD), best 20 m sprint time and RSA. The RSA test involved 12x20 m sprints departing every 20 s. When including the work performed during the time taken to decelerate, the test involved a work to rest ratio of approximately 1:3. Total sprinting time and the percentage decrement of repeated sprinting times were the two derived measures of RSA. The results indicate that the best 20 m sprint time was the only factor to correlate significantly with total sprinting time (r = 0.829, P < 0.001) and percentage decrement (r = -0.722, P < 0.01). VO2 max and AOD were not related to the total sprinting time or the percentage decrement that was produced by the RSA test. This was interpreted to signify that the phosphagen system was the major energy contributor for this test. PMID- 9732115 TI - Methods of the NSW Schools Fitness and Physical Activity Survey, 1997. AB - Physical fitness, participation in physical activity, fundamental motor skills and body composition are important contributors to the health and the development of a healthy lifestyle among children and youth. The New South Wales Schools Fitness and Physical Activity Survey, 1997, was conducted to fill some of the gaps in our knowledge of these aspects of the lives of young people in New South Wales. The survey was conducted in February and March, 1997 and collected data on a randomly-selected sample of students (n = 5518) in Years 2, 4, 6, 8 and 10. Measures were taken on body composition (height and weight, waist and hip girths, skinfolds), health-related fitness (aerobic capacity, muscular strength, muscular endurance, flexibility), fundamental motor skills (run, vertical jump, catch, overhand throw, forehand strike and kick), self-reported physical activity, time spent in sedentary recreation, and physical education (PE) classes. The methods are described to assist in the development of surveys of other populations and to provoke debate relevant to the development and dissemination of standard approaches to monitoring the fitness, physical activity habits and body composition of Australian children and youth. Finally, we offer comments on some of the strengths and limitations of the methods employed. PMID- 9732116 TI - The dynamic loading response of surfaces encountered in beach running. AB - The purpose of this study was to measure the response to dynamic loading of sand surfaces typically encountered in beach running. An instrumented drop test rig was constructed and used to guide a drop mass through impact with two surfaces (i) dry, uncompacted sand; and (ii) wet, compacted sand. Four drop masses (3.86, 7.24, 10.62 and 14.0 kg) were chosen and dropped from four different drop heights (100, 200, 300 and 400 mm) to represent the kinetic energies typically experienced during heelstrike in running. Accelerations were measured using a piezoelectric accelerometer and the trajectory of the drop head was measured using a displacement transducer. The following response variable were calculated for each trial: (i) peak impact force, (ii) mean impact force, (iii) impulse, (iv) total impact time, (v) rise time, (vi) fall time, (vii) maximum penetration, (viii) energy absorbed by the surface, and (ix) surface stiffness. Mean and peak impact forces were approximately 4 times greater for the wet surface while penetration, impact time and rise time were approximately 3-4 times greater for the uncompacted surface condition. The wet surface was also found to be 6 times stiffer than the uncompacted surface indicating the presence of water substantially altered surface compliance. Results are discussed in terms of their implications for performance and the potential for injury to athletes who run on these surfaces. PMID- 9732117 TI - Physical performance differences between weight-trained sprinters and weight trainers. AB - The present study tested and compared well-trained athletes who were performing low-velocity, high-force resistance training and sprint running training (ST) when recruited, with subjects who were performing low-velocity, high-force resistance training but not sprint training (NST) when recruited. Eleven male sprint runners (mean +/- SD; age = 19.0 +/- 1.4 yr: height = 182.0 +/- 4.7 cm: mass = 75.7 +/- 4.7 kg), and eight male weight-trained athletes who were not currently performing sprint training, or any other additional training, (mean + SD; age = 21.5 +/- 1.8 yr: height = 184.5 +/- 3.6 cm: mass = 78.4 +/- 4.6 kg) participated in the study; all subjects had a minimum of two years resistance training experience. Tests included 1. running speed (20 m time after a 50 m acceleration distance and 20 m acceleration time from a stationary start), 2. isokinetic hip flexor/extensor torque (and torque adjusted for body mass), angle of peak torque, time to reach peak torque and torque acceleration energy at low (1.05 rad x s(-1) [60 degrees x s(-1)), moderate (4.74 rad x s(-1) [270 degrees x s(-1)) and high (8.42 rad x s(-1) [480 degrees x s(-1)) speeds and 3. maximum squat lift. ST subjects produced more isokinetic hip extensor torque when adjusted for body mass at 4.74 rad x s(-1) (270 degrees x s(-1); p<0.05) and reached their peak torque faster (p<0.05). ST subjects also produced more hip flexor torque at 8.42 rad x s(-1) (480 degrees x s(-1); p<0.05), and torque per body mass at 4.74 rad x s(-1) (270 degrees x s(-1)) and 8.42 rad x s(-1) (480 degrees x s(-1); p<0.05) and reached peak flexor torque faster than NST subjects (4.74 rad x s(-1) [270 degrees x s(-1)], p<0.05; 8.42 rad x s(-1) [480 degrees x s(-1), p<0.01). Further, ST subjects performed better in tests of running acceleration over 20 m (p<0.02) and achieved a higher maximum running velocity after a 50 m acceleration distance (p<0.001). No significant differences were found in isokinetic strength at low (1.05 rad x s(-1) [60 degrees x s(-1)) velocities or in maximal squat lift strength. The results of the present study suggest that athletes who perform low-velocity, high force training concurrently with high-velocity training are superior in tests of isokinetic strength at high velocities when compared to athletes who only perform low-velocity, high force training. This may be due to training or genetic factors. PMID- 9732118 TI - The VISA score: an index of severity of symptoms in patients with jumper's knee (patellar tendinosis). Victorian Institute of Sport Tendon Study Group. AB - Symptoms of jumper's knee (patellar tendinosis) are not easily quantified and this may explain why there are no evidence-based guidelines for managing the condition. A simple, practical questionnaire-based index of severity would facilitate jumper's knee research and subsequently, clinical management. Thus we devised and tested the Victorian Institute of Sport Assessment (VISA) questionnaire. The brief questionnaire assesses (i) symptoms, (ii) simple tests of function and (iii) ability to play sport. Six of the eight questions are scored on a visual analogue scale from 0-10 with 10 representing optimal health. The maximal VISA score for an asymptomatic, fully performing individual is 100 points and the theoretical minimum is 0 points. We found the VISA scale to have excellent short-term test-retest, and inter-tester reliability (both, r>0.95) as well as good short-term (one week) stability (r=0.87). Mean (SD) of the VISA scores ranged from 95 (8) points in asymptomatic control subjects to 55 (12) points in patients who presented to a sports medicine clinic with jumper's knee and 22 (17) points in patients before surgery for chronic jumper's knee. Six- and twelve-months after surgery VISA scores returned to 49 (15) and 75 (17) points respectively, mirroring clinical recovery. We conclude that the VISA score is a reliable index of the severity of jumper's knee that has potential to aid clinicians and researchers. PMID- 9732119 TI - The influence of game location on athletes' psychological states. AB - The purpose of the study was to investigate the relationship between game location and precompetition psychological states. Male rugby players (N = 100) completed the Competitive State Anxiety Inventory-2 and the Profile of Mood States approximately 1 hr before a home and an away game. Repeated measures multivariate analysis of variance of mood and anxiety scores indicated significant differences between home and away locations. Participants scored higher on Vigor and Self-confidence, and lower on Tension, Depression, Anger, Fatigue, Confusion, Cognitive Anxiety, and Somatic Anxiety when competing at home. The findings support the proposal (Courneya & Carron, 1992) that psychological states are influenced by game location. PMID- 9732120 TI - Applying the Sports Medicine Australia pre-exercise screening procedures: who will be excluded? AB - Recently Sports Medicine Australia (SMA) and the Australian Association for Exercise and Sport Science (AAESS) developed guidelines for pre-exercise screening and supervision of fitness testing, based on the American College of Sports Medicine (ACSM) system. The procedure involves classifying individuals into one of three risk groups (apparently healthy, at higher risk, with known disease). Using data collected in a 1992 survey of 2298 Australian adults aged 18 78 years conducted by the Department of the Arts, Sport, the Environment and Territories (DASET), we calculated the percentage of the general population falling within each risk group and therefore exclusion rates (ie the proportion of subjects who, it is recommended, would require medical clearance prior to exercise or exercise testing). The analysis of data found that between 43-73% of males and 44-61% of females would require clearance. A cost analysis suggests that a rigorous application of the SMA-AAESS guidelines would cost between $250 million and $1.2 billion each year. On the basis of the results, suggestions for reviewing the guidelines have been proposed. PMID- 9732121 TI - Anthropometric profiles of elite triathletes. PMID- 9732122 TI - The guanylin and uroguanylin peptide hormones and their receptors. AB - Guanylin and uroguanylin are newly discovered, related peptides that activate common guanylyl cyclase signaling molecules and via 3', 5'-guanosine cyclic monophosphate regulate the activity of a variety of tissues and organs. Additionally, the message for both peptides is expressed in a variety of tissues and organs, including the intestinal tract and kidney, and thus may serve as part of a functional endocrine axis linking these two major organ systems in fluid/volume homeostasis. This manuscript reviews the discovery and nature of the guanylin and uroguanylin peptides, their actions on the intestinal mucosa and kidney, the distribution and molecular biology of the guanylyl cyclase C receptor, and explores the future directions of this rapidly developing, expanding field of inquiry. PMID- 9732123 TI - Characterization of the inner enamel epithelium in the enamel-free area based on the ability to secrete enamel protein demonstrated by in situ hybridization and immunohistochemistry. AB - Both the expression of amelogenin mRNA and secretion of amelogenin were investigated in rat molars by in situ hybridization and immunohistochemistry. Probes were designed by multiple-labeling of oligonucleotide probes for in situ hybridization. Amelogenin mRNA first appeared in differentiating ameloblasts of the distal region and some inner enamel epithelial cells of enamel-free area (EFA cells) of the second cusp at postnatal day 0. At the same time, amelogenin protein was detected in the extracellular matrix between dentin and differentiating ameloblasts and in some EFA cells of the second cusp. At postnatal day 1-3, amelogenin was expressed in the secretory ameloblasts, and in the matrix beneath these cells. Both amelogenin mRNA and amelogenin were detected in the EFA cells and their extracellular matrix. After postnatal day 5, amelogenin mRNA and amelogenin were detected in the secretory ameoloblasts and extracellular matrix in the enamel-forming region, respectively. At this time, amelogenin mRNA was not detected in the EFA cells, but a small amount of amelogenin was found in the matrix beneath the EFA cells. These findings suggest that EFA cells differentiate into amelogenin-secreting cells, i.e. ameloblasts, but that the secretion lasts for only a short period at the early stage of tooth development. PMID- 9732124 TI - Distribution and neuropeptide content of nitric oxide synthase-containing nerve fibers in arteries and conduction system of the rat heart. AB - The rat heart receives its blood supply not only from the coronary arteries but also from the accessory coronary arteries that supply mainly the atria. The distribution and chemical nature of nitric oxide synthase-containing nerve fibers in the conduction system as well as the coronary and accessory coronary arteries were investigated using immunohistochemistry and NADPH diaphorase (NADPH-d) histochemistry. A few NADPH-d-positive nerve fibers were observed mainly around the main trunk of the coronary arteries, while NADPH-d-positive fibers were found along the entire course of the accessory coronary arteries from their main branches to the arteriolar level. A double-staining method demonstrated that NADPH-d-positive fibers innervating both the coronary and accessory coronary arteries contained vasoactive intestinal polypeptide or neuropeptide Y. NADPH-d positive fibers were relatively abundant in the sinus node and penetrating bundle but were very sparse in the atrioventricular node and right bundle branch. No NADPH-d-positive fibers were detected in the left bundle branch. Some of the NADPH-d-positive fibers innervating the penetrating bundle exhibited distinct immunoreactivity to calcitonin gene-related peptide. These results suggest that nitric oxide may play a role as a neurotransmitter and/or neuromodulator in the neural control of the cardiac blood flow and impulse conduction. PMID- 9732126 TI - The sympathetic nerves of the parasellar region: pathways to the orbit and the brain. AB - Sympathetic nerves innervate targets in the orbit and the brain. They issue from the superior cervical ganglion and reach the parasellar region via the internal carotid nerve. Information on their further parasellar course and distribution is scant and contradictory. In this study the parasellar sympathetic pathways of 30 human infants and 6 human fetuses were investigated by microdissection and histologically. A common parasellar sympathetic trunk, which reunites all the nerve fibers emanating from the lateral and medial internal carotid plexus, is described as well as its further divisions. It was found that the posterior knee of the infant carotid siphon is free of large sympathetic nerve bundles. In addition a ganglion is described, which is situated in the parasellar adipose body. It is reached by nerve fibers coming from the parasellar sympathetic pathways. Fibers that issue from this ganglion join the periorbita and the orbital muscle of Muller. These anatomical facts are of immediate importance for preventing nerve damage during cavernous sinus surgery. Furthermore, the study improves the anatomical knowledge of the parasellar region and suggests a new concept for the innervation of the orbital muscle. PMID- 9732125 TI - Mitochondrial density of ventral horn neurons in the rat spinal cord. AB - Mitochondrial density in neurons of the dorsolateral region of the ventral horn at the L5 spinal cord segment in rats was examined using electron microscopy. The gamma motoneurons had a higher density of mitochondria (25.1 +/- 4.2%, n = 19) in the cytoplasm compared to the alpha motoneurons which had a mitochondrial density of 19.4 +/- 4.5% (n = 38). An inverse relationship between cell body size and mitochondrial density was found for alpha (n = 38) and alpha plus gamma (n = 57), but not for gamma (n = 19), motoneuron populations. The higher densities of mitochondria in the smaller neurons correspond well with their metabolic properties since the smaller neurons have the highest oxidative enzyme activities. PMID- 9732127 TI - Postnatal development of the cruciate ligament insertions in the rat knee. morphological evaluation and immunohistochemical study of collagens types I and II. AB - The postnatal structural remodelling and calcification patterns in the insertions (entheses) of both cruciate ligaments were studied in a rat model with histology and immunohistochemical analysis of collagens types I and II. In the neonate, both ligaments which labelled only for type I collagen attached to epiphyseal cartilage which solely labelled for collagen type II. The entheses calcified between days 20 and 35, and a subchondral bone plate formed under the entheses between days 30 and 55. Thus, within a period of 35 days the tissue to which the ligaments attached increased multifold in stiffness. Interestingly, the process of enthesial calcification and formation of compact bone did not happen simultaneously in both ligaments, not even synchronous at both ends of the same ligament or within a single insertion. This asynchronous calcification of the different knee ligament insertions may make the sudden change in mechanical environment at the entheses less dramatic for the ligaments and knee joint surfaces as anticipated from mechanical models. In addition, a fibrocartilaginous tissue, rich in collagen type II, formed in the ligament at a time when the epiphyseal cartilage was replaced by bone, and grew wider with time. The interposition of a fibrocartilaginous zone in the insertion may diminish the sudden change in stiffness between ligament soft tissue and hard bone. PMID- 9732128 TI - A calculation of the forces acting on the human acetabulum during walking. Based On in vivo force measurements, kinematic analysis and morphometry. AB - Information about the loading of the human acetabulum during walking is necessary for a functional understanding of the morphology of the pelvic girdle and the hip joint as well as for the optimization of endoprosthetic therapy in osteoarthritis. For this purpose, experimental data of the forces acting on the femur in walking taken from the literature [Bergmann et al.: J. Biomech. 1993;26: 969-990] were combined with our own kinematic and morphometric data, to transform the force vectors from the femoral into a pelvic and an acetabular frame. During the walking cycle, the resultant force vector takes a rather constant course relative to the pelvis and its orientation seems to be highly regulated to act within a small range of angles. Only small deviations occur from the angles against the vertical which the resultant peak force forms in the frontal plane (F = 11 degrees, medially orientated) and in the sagittal plane (S = 5 degrees, ventrally orientated). The experimental results form the basis for a model of the incongruous hip joint as an elastic joint, the femoral head being centered between compliant elements. PMID- 9732129 TI - Effect of temperature on the transient evoked and distortion product otoacoustic emissions in rats. AB - In order to study the energy dependence of the cochlear amplifier, transient evoked otoacoustic emissions (TEOAEs) and distortion product otoacoustic emissions (DPOAEs) were recorded in rats during gradual cooling to 27 degrees C and heating to 40 degrees C. In the range 33-39 degrees C, the TEOAEs and DPOAEs were maximal in amplitude and almost insensitive to temperature. However, they were significantly depressed (reversibly) at higher and lower temperatures. Intensity functions were plotted at 37, 27 and 40 degrees C for both types of oto acoustic emissions. At 37 degrees C intensity functions were nonlinear, with a notch at mid-intensity regions. At 27 degrees C, the magnitudes were depressed more at the lower intensities and threshold elevations were observed. As a result, the intensity functions were more linear and the notch was no longer seen. This result provides further evidence for a more active, energy-dependent component of the otoacoustic emissions at lower intensities for both TEOAEs and DPOAEs. The cooling probably affects the lower intensity otoacoustic emissions by inducing a depression in the endocochlear potential, by reducing the motility of the outer hair cells and by introducing a small conductive hearing loss. PMID- 9732130 TI - Basal cochlear lesions result in increased amplitude of otoacoustic emissions. AB - We have measured the changes in transient otoacoustic emissions (TEOAEs) and distortion product otoacoustic emissions (DPOAEs) during and after ototoxic amikacin treatment in an animal (chinchilla) model. TEOAE and DPOAE were recorded from 6 adult chinchillas over a 6-week time course starting just before a 5-day or 7-day treatment period with amikacin sulphate (400 mg/kg/day, i.m.). After final recordings, cochlear morphology was assessed by scanning electron microscopy. Generally, both DPOAE and TEOAE amplitudes change during and after treatment in a systematic fashion. High-frequency components change first, followed by lower-frequency components. We note that there is often a long latency to the onset of changes in otoacoustic emissions (OAE), and that these changes can continue for weeks after treatment. Most importantly we report that when the basal region of the cochlea is damaged in the frequency region above the OAE recording bandwidth (0.6-6 kHz for TEOAE; 1-6.7 kHz for DPOAE), we often find an increase in OAE amplitudes. More specifically, we note that as a cochlear lesion progresses apically, there is often a transient increase in a frequency specific OAE before it reduces or is lost. Our results suggest that the increase in OAE amplitudes precedes the expression of detectable cochlear pathology. PMID- 9732131 TI - Transient-evoked and 2F1-F2 distortion product oto-acoustic emissions in dogs: preliminary findings. AB - Transient (click)-evoked oto-acoustic emissions (TEOAEs) and distortion product oto-acoustic emissions (DPOAEs) were recorded in a feasibility study in 7 healthy mixed-breed dogs using the ILO 92 OAE analyser (Otodynamics, Hartfield, UK). Five dogs were found to have normal hearing in both ears and 2 dogs in the left ear only following otoscopy, tympanometry and auditory brainstem response audiometry. Twelve sets of TEOAEs (click-evoked) to 80 dB peSPL click stimulus and 9 sets of DPOAEs (2F1-F2) to 8 different stimulus levels of the primary tones (L1/L2) were collected at 11 test frequencies (F2) in these normal-hearing dogs. TEOAEs were successfully recorded in 11 of the 12 ears using the default user setting and in all 12 ears using the quickscreen program. DPOAEs were successfully recorded in all 9 ears tested. While the TEOAEs parameters matched those for humans, the average signal-to-noise ratio of DPOAEs was considerably higher in the dogs. Stimulus levels at 55/55, 55/45 and 55/35 dB SPL were demonstrated to produce DPOAEs that seem to reflect the active dynamic status of the outer hair cell system. Postmortem DPOAEs at these stimulus levels and TEOAEs at 80 Db peSPL could not be elicited 5 min following euthanasia of dogs. However, DPOAEs could still be recorded albeit with reduced amplitude at stimulus levels where L1 > 55 dB SPL. The results suggest that TEOAEs and DPOAEs in dogs have the potential to provide valuable insights into their mechanisms of generation, and the specific role and behaviour of outer hair cells of the cochlea in certain pathological conditions, particularly in drug-induced ototoxicity, in humans. PMID- 9732132 TI - Effects of age, gender and ear side on SOAE parameters in infancy and childhood. AB - We investigated 267 infants and children aged 9 days to 16.8 years to study the spontaneous otoacoustic emission (SOAE) data prevalence, number per ear, level and frequency as a function of growth. Dependence on age, gender and ear side was statistically analyzed using the method of generalized estimation equations. Except in the 1st year of life, SOAE prevalence per ear and SOAE number per ear decreased significantly with increasing age. Both SOAE parameters were significantly higher in female than in male subjects, with gender difference of SOAE prevalence per ear being more distinct in the 1st year of life. Although a clear ear side effect on SOAE prevalence per ear could already be seen in ears of female children in this age group, only SOAE number per ear was significantly higher in right ears than in left ears from the 1st year of life on. Except in the first 12 months, SOAE level and SOAE frequency decreased significantly with increasing age. Neither a significant gender difference nor a significant ear side difference could be determined. Our results found in infancy and childhood are discussed within the framework of the current literature. PMID- 9732133 TI - Clinical applicability of transient evoked otoacoustic emissions: identification and classification of hearing loss. AB - The study aimed at the development of a clinically applicable methodology that could: (1) discriminate transient evoked otoacoustic emission (TEOAE) recordings from normal hearing or hearing impaired individuals; (2) classify the nature of the hearing loss as conductive or as cochlear, and (3) define clear-cut TEOAE clinical criteria. A classification algorithm based on a multivariate discriminant analysis of fast Fourier transform data from recordings evoked by click stimuli of 50 +/- 2, 62 +/- 2, 68 +/- 2 and 80 +/- 2 dB SPL was used to discriminate 302 normal subjects from 383 subjects suffering from mild to moderate hearing losses. The best discriminant model (QDF80) produced a sensitivity of 93.8% and a specificity of 79.4%. When extra correlation criteria were serially applied to the classification outcome, the specificity was increased to 85.3%, but the sensitivity was marginally decreased to 91.7%. The classification of the correctly identified hearing-impaired cases yielded 93.8% identification of conductive and 75.1% identification of cochlear cases. A sensitivity analysis of the misclassified hearing-impaired cases suggested that the TEOAE spectra are well correlated with the 2-kHz but poorly correlated with the 4-kHz octave frequency. PMID- 9732134 TI - Classification of patients with Meniere's disease using otoacoustic emissions. AB - Otoacoustic emissions (OAEs) were studied in patients with Meniere's disease in order to assess their usefulness for distinguishing between different stages of the disease. An accurate classification of Meniere's patients is expected to allow optimalization of the treatment for each particular stage. Click-evoked and distortion product OAEs were evaluated in both ears for 70 Meniere's patients. Based on these measurements, Meniere's patients can be divided into four different categories. In patients with small hearing losses OAEs are found, whereas patients with pure-tone thresholds larger than 60 dB exhibit no OAEs at all. In the intermediate range (30- to 60-dB thresholds) two categories of patients are distinguished: patients with relatively large emissions and patients without measurable emissions. These findings suggest the presence of various stages in the pathophysiological mechanism involved in Meniere's disease. Additionally, Meniere's patients with contralateral ears with normal thresholds have significantly smaller emissions than normal-hearing adults. This observation could represent a very early manifestation of bilateral Meniere's disease, which cannot be detected by other diagnostic methods. PMID- 9732143 TI - Stenotrophomonas (Xanthomonas) maltophilia: in vitro susceptibility to selected antimicrobial drugs, single and combined, with and without defibrinated human blood. AB - Sixteen selected isolates of Stenotrophomonas maltophilia varied in susceptibility to the combined phagocytic/serum bactericidal activity of fresh defibrinated human blood (65 vol%). Four representative isolates (X1, X11, X25, and X50), which differed in susceptibility to cefepime, ceftazidime, rifampin, and timentin, were subjected to checkerboard microtiter broth dilution tests involving combinations of cefepime plus timentin, ceftazidime plus ofloxacin, cotrimoxazole plus timentin, rifampin plus polymyxin B, and rifampin plus polymyxin B nonapeptide; all combinations yielded additive or synergistic effects against all four strains. Unexpectedly, the combination of cefepime plus timentin was bactericidally active against the two cefepime-resistant isolates. This finding was substantiated by blood/broth plus combined antimicrobial drug assays. Cefepime plus timentin effectively killed all four test strains. Ofloxacin combined with ceftazidime was bactericidally active against the test strains, including two isolates (X11, X50) with intermediate ofloxacin sensitivity. Cotrimoxazole plus timentin in blood, but not in broth, was bactericidal for the timentin-resistant isolate X25. As expected, various triple combinations of chemotherapeutic agents in blood and broth revealed polymyxin B plus rifampin, regardless of the third combination partner, to exert bactericidal activity against all test strains. Similarly, rifampin combined with ofloxacin and ceftazidime was bactericidally active in blood and broth. The observation that timentin combined with cefepime was effective against cefepime-resistant strains of S. maltophilia might prove of clinical relevance with regard to chemotherapy of nosocomial infections due to multiple-antibiotic resistant strains of this opportunistic pathogen. PMID- 9732144 TI - In vitro activity of piperacillin/tazobactam against 615 Pseudomonas aeruginosa strains isolated in intensive care units. AB - From May 1996 to September 1997, 615 Pseudomonas aeruginosa strains isolated from patients in intensive care units collected from different Italian laboratories were studied. The susceptibility of piperacillin/tazobactam, in comparison with other antipseudomonal antibiotics, to their NCCLS breakpoints was evaluated: amikacin 79. 6%, carbenicillin 67.0%, ceftazidime 73.4%, ciprofloxacin 55.8%, imipenem 64.1%, piperacillin 88.1%, piperacillin/tazobactam 92.4% and ticarcillin/clavulanic acid 69.0%. Seventy-three strains were selected because of their resistance to piperacillin and the mechanisms underlying such a resistance were investigated. Isoelectric focusing and hydrolysis assays revealed the presence of 15 plasmid-mediated beta-lactamases. Chromosomal beta-lactamase derepression was demonstrated in 34 isolates. The remaining 24 piperacillin resistant strains did not produce beta-lactamases and an 'intrinsic mechanism' of resistance was inferred. The piperacillin/tazobactam combination restored resistance in 25 piperacillin strains. Nine of these were derepressed for chromosomal beta-lactamase, 8 showed impermeability and 8 showed plasmid enzymes. PMID- 9732145 TI - Effect of different beta-lactams in combination with beta-lactamase inhibitors in the presence or absence of tobramycin against some enterobacteriaceae producing extended-spectrum beta-lactamases. AB - Extended-spectrum beta-lactamase (ESBL) production among members of the family Enterobacteriaceae generally involves resistance to oxyimino-cephalosporins and monobactams, while implying different susceptibility profiles with other antimicrobial agents. We have investigated the activity of some beta-lactam antibiotics, alone or in double and triple combinations with beta-lactamase inhibitors, in the presence or absence of tobramycin (TOB), against some Enterobacteriaceae producing ESBL by means of time-kill curves. Antimicrobials employed were ceftazidime (CAZ), cefotaxime (CTX), TOB, ampicillin (AMP), ampicillin-sulbactam (ASL), amoxicillin (AML), amoxicillin-clavulanic acid (AMC), piperacillin (PIP) and piperacillin-tazobactam (TZP), at 1/4 minimum inhibitory concentration for susceptible strains and at achievable serum concentrations for resistant strains. Only the combinations CTX-ASL, CAZ-ASL and PIP-ASL were synergistic, both at 6 and 24 h, on some strains of Klebsiella species. PMID- 9732147 TI - Effect of cefamandole, cefuroxime and cefoxitin on yeast fecal flora of surgical patients. AB - Aim of the present study was to evaluate the effect of cefamandole, cefuroxime and cefoxitin on the level of gastrointestinal (GI) colonization by Candida albicans in humans. Twenty-eight adult patients received one of these three cephalosporins for 10 days, as treatment of infection, and were studied prospectively. Quantitative stool cultures for yeasts were performed immediately before, at the end, and 1 week after discontinuation of treatment. All three antibiotics caused an increase of the yeast concentration in the fecal flora. The increase caused by cefoxitin was the highest (2.5 log10 CFU/g of stool). Our results suggest that the cephalosporins tested cause minor increases of the colonization of the GI tract by C. albicans. PMID- 9732146 TI - Comparative in vitro activity of vancomycin and other antimicrobial agents against methicillin-resistant staphylococcus aureus and enterococcus faecium in the Tohoku district of Japan. AB - Susceptibility patterns of methicillin-resistant Staphylococcus aureus (MRSA) and Enterococcus faecium obtained from various hospitals of the Tohoku district were documented. MICs of 6 antimicrobial agents against a total of 480 strains (380 strains were MRSA and 100 were E. faecium) were estimated. All MRSAs were susceptible to vancomycin, teicoplanin and quinupristin/dalfopristin, but all of them were resistant to ampicillin and benzylpenicillin. None of the E. faecium strains were found to be resistant to vancomycin, teicoplanin and quinupristin/dalfopristin. Excluding these, almost all strains of E. faecium were resistant to the remaining drugs. These data suggest that despite the emergence of vancomycin resistance to E. faecium in Europe and in the United States, vancomycin, teicoplanin and quinupristin/dalfopristin will nevertheless provide effective bactericidal activity in the Tohoku area of Japan. PMID- 9732148 TI - In vitro activities of E1101, a novel oral cephalosporin, against bacteria causing infections in obstetric and gynecological patients. AB - E1101 is a new oral cephalosporin with a broad spectrum of antibacterial activity. It inhibited more than 90% of clinical isolates of Streptococcus agalactiae, Escherichia coli and Peptostreptococcus magnus at the concentration of 3.13 mg/l. E1101 was the most active agent against S. agalactiae and E. coli. Since none of the compounds was sufficiently active against the Bacteroides fragilis and Prevotella bivia isolates, they are not appropriate in the treatment of patients with infections caused by these organisms. The results of this study suggest that, subject to confirmation by clinical trials, E1101, in combination with an agent with reliable activity against anaerobic bacteria, is suitable as empirical therapy of patients with obstetric and gynecological infections. PMID- 9732149 TI - Effect of a 17-member azalide on tumor cell growth. AB - The action of the 17-member azalide 4'-demycarosyl-20-deoxo-20-(di-N-benzylamino) 8a-aza-8a-homotyl osin was examined in vitro using five different human cell lines: laryngeal carcinoma (Hep2), pancreatic carcinoma (MiaPaCa2), breast carcinoma (MCF7), neuroblastoma, and normal diploid fibroblast (Hef522). After exposure, the cell growth was arrested, and morphological changes occurred in a dose-dependent manner. At a concentration of 10(-4) M the azalide completely inhibited the growth of all cell lines examined and induced morphological changes such as cell shrinkage, chromatin condensation, and DNA fragmentation. These features point to apoptosis. PMID- 9732150 TI - Bactericidal effect of levofloxacin on strains with equal susceptibility in an in vitro urinary bladder model. AB - The in vitro bactericidal effects of therapeutic concentrations of levofloxacin (LVFX) on clinical isolates of Pseudomonas aeruginosa and Enterococcus faecalis were assessed. Starting from 10(7) CFU/ml, the bacterial count was determined serially during the exposure to LVFX present at concentrations automatically simulated to its clinically achievable urinary levels at the recommended dosage (100 mg 3 times daily). An isolate of P. aeruginosa for which the MIC of LVFX was 16 microgram/ml was completely eradicated after 10 h of exposure to LVFX. On the other hand, 40 h of exposure to LVFX were required for complete eradication of an isolate of E. faecalis with equal susceptibility, although the bacterial count was rapidly reduced to 10(2) CFU/ml in the early phase of the exposure. It was concluded that LVFX exerts a more potent bactericidal effect on P. aeruginosa than on E. faecalis despite a similar susceptibility of these organisms to LVFX. PMID- 9732151 TI - Ceftriaxone monotherapy in the treatment of low-risk febrile neutropenia. AB - Febrile neutropenia in patients who have undergone chemotherapy is usually treated with a combination of broad-spectrum antibiotics. There are no exactly defined protocols for single-agent treatment because a clear definition of low risk febrile neutropenia is lacking. This paper examines the safety and efficacy of once-daily ceftriaxone in 376 cases. MATERIAL AND METHODS: In a prospective observational study carried out between February 1992 and January 1996, 959 febrile episodes at 48 hospitals were recorded. Inclusion criteria were neutropenia (absolute neutrophil count, ANC <1,000/ microl) with fever (>/=38.5 degreesC) or a C-reactive protein concentration >1 mg/dl and suspected infection. Nine hundred and one episodes (acute leukemia n = 396, lymphoma n = 220, solid tumors n = 272 and other disorders n = 13) in 828 patients aged between 1 and 97 years were analyzed, of which 876 episodes were evaluable for response. All patients initially underwent empirical treatment with ceftriaxone (adults: 2 g/day; children: 80 mg/kg/day), either alone (376) or in combination with other agents (525). RESULTS: The mean ANC was 423/ microl (SD +/- 316) and the median duration of neutropenia 10 days. Of the 363 episodes treated initially with ceftriaxone alone, 70.8% responded versus 56.9% in the combination therapy group. The favorable response to the initial monotherapy treatment was explained by a low-risk population in the monotherapy group. A KI >6 (p < 0.0001), ANC >/=500/ microl (p = 0.0001) and a duration of ANC <5 days (p < 0.05) were significantly more frequent in the monotherapy arm and were predictive of lower risk at the commencement of treatment. CONCLUSION: Ceftriaxone is effective in febrile neutropenia. Treatment with ceftriaxone alone was safe and highly effective in low-risk patients. Single-agent regimens appear to be a suitable treatment option in low-risk febrile neutropenia. PMID- 9732152 TI - Antimicrobial prophylaxis in laparoscopic and conventional cholecystectomy . Conclusions of a large prospective multicenter quality assurance study in Germany. AB - BACKGROUND: Postoperative infection following cholecystectomy poses a significant threat to recovery, with major cost repercussions. Though antimicrobial prophylaxis is commonly practiced, its value - particularly in laparoscopic cholecystectomy - has not yet been adequately documented. METHOD: In a prospective multicenter quality assurance study in 28 German hospitals, an analysis of data collected on 4,477 patients undergoing conventional (n = 1,349) or laparoscopic (n = 3,128) cholecystectomy was performed; 2,217 patients received and 2,260 did not obtain perioperative antibiotic cover. RESULTS: Postoperative infections occurred in a total of 136 patients, with infection rates of 5.0% in those without prophylaxis, 0.8% in those on ceftriaxone, and 1.2% in those on other antibiotic regimens. Patients receiving prophylaxis fared significantly better than those with no prophylaxis in terms of the rate of postoperative wound infections, chest infections, other complications, reoperation and mortality. CONCLUSION: Neither laparoscopic nor conventional open cholecystectomy should be performed without adequate perioperative antimicrobial prophylaxis in future, especially since such measures also reduce hospital stay and hence the costs. PMID- 9732154 TI - Phlebology: an essential part of dermatology. PMID- 9732153 TI - Comparative study on the effectiveness of antifungal agents in different regimens against vaginal candidiasis. AB - OBJECTIVE: A study was carried out to compare three treatment regimens for vaginal candidiasis. METHODS: A total of 150 women with clinical and mycological evidence of vaginal candidiasis were randomized to receive daily a 200-mg dose of oral itraconazole for 3 days (50 women), a single oral 150 mg dose of fluconazole (50 women), or daily 100 mg dose of intravaginal clotrimazole for 6 days (50 women). They were assessed at 5-15 days (short-term assessment) and again at 30 60 days (long-term assessment) after discontinuation of the treatment. RESULTS: At the short-term or long-term assessment, Candida species were completely eradicated from the vagina in 80 or 74% in the 3-day oral itraconazole group, 76 or 70% in the single oral fluconazole group, and 72 or 60% in the intravaginal clotrimazole group, respectively. The rates of clinical effectiveness were 92 or 88% in the 3-day oral itraconazole group, 80 or 76% in the single oral fluconazole group, and 72 or 58% in the intravaginal clotrimazole group, respectively. Treatment-related side effects were not found in any group. CONCLUSIONS: Our study suggests that the treatment of vaginal candidiasis with oral itraconazole or oral fluconazole would be effective and that an oral itraconazole or fluconazole therapy might be one choice in the treatment of vaginal candidiasis. PMID- 9732155 TI - Cutaneous mastocytosis in adults. evaluation of 14 patients with respect to systemic disease manifestations. AB - BACKGROUND AND OBJECTIVE: Systemic mastocytosis is a rather rare disorder involving the skin and several other organs. The aim of this study was to analyse the extent of extracutaneous manifestations in 14 adult patients who presented with prominent cutaneous involvement within the last 5 years. RESULTS: The cutaneous lesions were clinically diagnosed as telangiectasia macularis eruptiva perstans in 2 patients, urticaria pigmentosa of varying extent in 11 and diffuse erythrodermic mastocytosis in 1 patient. All patients had extracutaneous manifestations with involvement of one additional organ system in 6/14 cases, two in 5/14 and three in 3/14. Ten out of 14 patients suffered from generalized pruritus, and 11/14 reported mild wheal formation, while 3/14 with multi-organ involvement mentioned recurrent flushing episodes. The gastro-intestinal tract was involved in 8/14 cases with an increase in gastric and colon mucosal mast cells in 5/8 cases and gastroduodenitis in 2. Bone marrow involvement was seen in 7/13 patients, hepatosplenomegaly in 2, anaemia in 2 and thrombocytopenia in 3. The disease had a duration of 0.5-32 years, clinical symptoms remaining basically unchanged. Malignant transformation was not seen; only 1 patient developed myelodysplastic syndrome within 2 years after the first cutaneous lesions. CONCLUSIONS: Our study shows that extracutaneous involvement should be carefully considered in adult patients with cutaneous mastocytosis. Systemic multi-organ mast cell disease in adults is a long-lasting disorder with recurrent episodes of varying clinical symptomatology. However, the disease shows rather slow progression, and malignant transformation is rare. Satisfactory management is achieved by symptomatic oral drug intake. PMID- 9732156 TI - Expression of stem cell factor in the lesional skin of systemic sclerosis. AB - BACKGROUND AND OBJECTIVE: Mast cells are thought to play an important role in the pathogenesis of systemic sclerosis (SSc), although their significance is still unknown. As mast cells are increased in number in the lesional skin of the early stage of scleroderma, we addressed the question whether expression of stem cell factor (SCF), a mast cell growth factor, is upregulated in the lesional skin. METHODS: Immunohistochemical analysis of SCF was performed in paraffin-embedded skin sections from the lesions of 18 patients with SSc (13 in the edematous and 5 in the sclerotic phase) and from normal skin of 5 subjects. SCF messenger RNA expression was also examined on cultured fibroblasts derived from SSc by using reverse-transcriptase polymerase chain reaction. RESULTS: Immunostaining showed focal expression for SCF in fibroblast-like spindle-shaped cells, mast cells, keratinocytes and endothelial cells. SCF-positive spindle-shaped cells were significantly increased in the reticular dermis in the early edematous stage of SSc (33.7 +/- 13.0/mm2) as compared with normal skin (18.3 +/- 4. 2/mm2; p <0.05); however, there was no significant difference between the sclerotic phase (18.9 +/- 6.9/mm2) and normal skin. mRNA expression of SCF was detected both in cultured scleroderma-derived and normal skin-derived fibroblasts. CONCLUSION: These results may suggest that SCF plays a role in the fibrotic process in early stage SSc. PMID- 9732157 TI - The acute effect of smoking on cutaneous microcirculation blood flow in habitual smokers and nonsmokers. AB - BACKGROUND: Smoking is known to be a primary cause of chronic harmful effects on the vascular system. It also induces some acute effects on the coronary circulation and on the ophthalmic artery. OBJECTIVE: The aim of our study was to assess the effect of smoking a single cigarette on cutaneous blood flow in habitual smokers as well as in nonsmokers. METHODS: A laser Doppler flowmeter was used to perform measurements of cutaneous microcirculation. Flowmetric data were recorded (i) before smoking, (ii) inhaling from an unlighted cigarette, (iii) during cigarette smoking, (iv) 2 and (v) 5 min after smoking. RESULTS: We could show that smoking a single cigarette acts on the cutaneous microcirculation reducing blood flow in both groups of subjects (38.1% reduction in smokers and 28.1% reduction in nonsmokers; p <0.01). Interestingly, the recovery phase is faster in nonsmoker subjects than in smoker ones; in fact, the recovery is complete 2 and 5 min after cigarette smoking in nonsmokers and in smokers, respectively. CONCLUSION: Smoking a single cigarette decreases the cutaneous blood flow in habitual smoker as well as in nonsmoker subjects. Moreover, the slower recovery phase of smokers suggests that their microcirculation become inured to smoke. PMID- 9732158 TI - Prolactin: does it have a role in the pathogenesis of psoriasis? AB - BACKGROUND: The aetiopathogenesis of psoriasis is still not fully understood. Recently, it has been reported that prolactin (PRL) exerts a proliferative effect on human keratinocytes in vitro. PRL may, therefore, play an important role in the pathogenesis of psoriasis. OBJECTIVE: To assess the serum PRL level in patients with psoriasis vulgaris (PV). METHODS: Serum levels of PRL were estimated in 12 patients with PV (age: 11-45 years with mean +/- SD 30.4 +/- 10.2 years; sex: 7 males, 5 females) and the results were compared with those in 9 patients with atopic dermatitis (age: 15-47 years with mean +/- SD 28.1 +/- 11.9 years; sex: 4 males, 5 females) and 20 normal control subjects (age: 16-45 years with mean +/- SD 36.1 +/- 11.9 years; sex: 15 males, 5 females). RESULTS: Serum PRL in PV (mean +/- SD 25.8 +/- 16.1 ng/ml) was significantly higher compared to those in atopic dermatitis (mean +/- SD 9.1 +/- 4.7 ng/ml) and normal control subjects (mean +/- SD 10.3 +/- 5.3 ng/ml; ANOVA --> p = 0.0008). Three patients with PV (2 males and 1 female with ages of 35, 40 and 11 years, respectively) had the highest serum levels well above the normal range but they were <100 ng/ml, the minimum limit for the diagnosis of prolactinoma (chi2 test --> p <0.025). CONCLUSION: Since PRL belongs to the growth hormone family, its raised serum level may have a role in the hyperproliferation of kerationocytes in vivo, the hallmark of the psoriasis disease process. PMID- 9732159 TI - Ki-67 immunostaining of normal human epidermis: comparison with 3H-thymidine labelling and PCNA immunostaining. AB - BACKGROUND: The size of the germinative growth fraction (i.e. the number of actively proliferating germinative cells) of normal human epidermis is still a subject of debate. Ki-67 antigen and PCNA, an auxiliary protein of d-polymerase, are considered as markers of the growth fraction when used under optimal conditions. METHOD: In the present work, we have compared Ki-67 expression (detected with MIB1 antibody) with PCNA expression (detected with PC10 antibody) in biopsies of normal human epidermis fixed in neutral formalin and using antigen retrieval by microwave processing. To obtain additional information, such as the percentage of cells in S phase, biopsies were also incubated in 3H-thymidine before immunostaining. RESULTS: Before microwave treatment, 8% of the basal cells were positive for MIB1 antibody and 7.8% were positive for PC10 antibody. The 3H thymidine labelling index was 2.8%. The proportion of MIB1-positive cells rose to 19% after antigen retrieval by microwave processing. In the same way, the 3H labelling index rose to 9%. In contrast, PC10 became positive in all epidermal nuclei. CONCLUSION: These results suggest that the growth fraction of the germinative cell population of normal human epidermis is not larger than 20% and is composed of cells with a short cell cycle time. PMID- 9732161 TI - Is aberrant mammary tissue a marker for chronic alcoholism or kidney-urinary tract malformations? AB - BACKGROUND: Numerous publications describe the relationship between aberrant mammary tissue (AMT) and kidney-urinary tract malformations, individual/ familial alcoholism and sense organ disorders. PATIENTS AND METHODS: We investigated these possible associations and reviewed 72 cases observed in our Department during the past 3 years: 30 men and 42 women, 17 of them with bilateral AMT (7 men and 10 women) and 25 patients from 9 families. Diagnosis was made according to Kajawa's classification. A detailed family history was performed asking for individual or familial alcoholism, especially in the mother, in addition to blood tests and ultrasonographic examination of the abdomen and the kidneys. RESULTS: We only found 1 family history of alcoholism in 3 families, but in the father, never in the mother or the affected subject. No congenital/ hereditary nephrourinary defects or sense organ disorders were found. CONCLUSIONS: We believe that in our population AMT is not a marker for alcoholism, kidney-urinary malformations nor sense organ disorders. PMID- 9732160 TI - Physiopathogenic investigations in a case of familial stiff-skin syndrome. AB - BACKGROUND: Stiff-skin syndrome (SSS) is a rare cutaneous syndrome characterized by stony-hard skin and limitation of joint mobility. Its cause is still unknown. OBJECTIVE: Biological investigations were performed in a new case of SSS. METHODS: Collagen production and DNA biosynthesis were studied from fibroblast culture. Proinflammatory cytokines (TNF-alpha, IL-6 and TGF-beta2) were measured in the patient's serum. Results were compared with pathological findings. RESULTS: Collagen production and DNA biosynthesis were normal whereas the level of circulating cytokines was high. Histological examination of the skin showed mild fibrosis in the dermis whereas the fascia was not thickened. CONCLUSION: Our clinical and biological findings suggest that in this case, cutaneous changes may be related to an inflammatory process rather than to a primary fibroblast defect or a fascial abnormality as previously hypothesized. PMID- 9732162 TI - Flow cytometry analysis of peripheral blood lymphocytes from patients with bullous pemphigoid. AB - BACKGROUND: Bullous pemphigoid (BP) is an autoimmune disease observed in elderly subjects in which alteration of peripheral blood lymphocyte subset repartition has been described. OBJECTIVE: To evaluate peripheral blood lymphocyte subsets from BP patients and identify immunologic changes that can be assigned either to the autoimmune disease or to the ageing process. METHODS: Peripheral blood lymphocytes from 24 patients with BP and 24 age- and sex-matched healthy individuals were analyzed by flow cytometry and these results were compared to those obtained from young adults. RESULTS: This study showed: (i) the presence of T-cell lymphopenia with a decreased number of CD8+ and gdgammadelta T lymphocytes in BP patients compared with younger adult controls; (ii) a high dispersion of the results in the BP population; (iii) the absence of a significant difference of lymphocyte subset levels between BP patients and age- and sex-matched controls. CONCLUSION: This flow cytometry study suggests that the modifications of lymphocyte subpopulation values observed in BP patients are related to immunologic changes induced by age in elderly people. PMID- 9732163 TI - Perfluoropolyethers in the prevention of irritant contact dermatitis. AB - BACKGROUND: Perfluoropolyethers (PFPEs) are non-reactive perfluorinated (no hydrogen atoms) liquid polymers that showed promise as ingredients of protective preparations in occupational dermatology. OBJECTIVE: The aim of this study was to investigate if oil-in-water (o/w) emulsions containing 0.5, 1.0, 2.0 and 4.0% PFPEs (molecular weight 6,250 D) prevent epidermal barrier disruption induced by a repetitive irritation test (RIT). METHODS: PFPE-containing o/w emulsions and the emulsion base alone were evaluated against a set of 4 irritants [10% sodium lauryl sulphate (SLS), 0.5% sodium hydroxide (NaOH), 15% lactic acid (LA), and undiluted toluene (TOL)] in the RIT. Ten subjects were tested on the paravertebral skin of the mid-back. Irritation was assessed by visual scoring, transepidermal water loss (TEWL) and colorimetry. RESULTS: Both the emulsion base and all PFPE-containing preparations significantly suppressed irritation by SLS and NaOH. Against LA and TOL, only the 4% PFPE-containing preparation was significant as assessed by TEWL. CONCLUSION: A 4% PFPE-containing o/w emulsion significantly inhibits irritation due to a wide spectrum of hydrophilic and lipophilic irritants. o/w emulsions with lower concentrations of PFPE are also effective against SLS and NaOH, but this is also true for the cream base alone. Therefore, further studies are necessary to evaluate if a minimum of 4% PFPE in the base provides additional benefit. PMID- 9732164 TI - Comparison of the effects of cetirizine and ebastine on the skin response to histamine iontophoresis monitored with laser Doppler flowmetry. AB - BACKGROUND: The administration of histamine with iontophoresis is an alternative method to skin prick tests or intradermal injections. Skin reactions obtained with this method can be recorded with laser Doppler flowmetry (LDF) and previous studies with this method have shown histamine-induced laser Doppler changes in the wheal area. OBJECTIVE: In order to compare the influence of two H1 receptor antagonists (cetirizine 10 mg vs. ebastine 10 mg) on the skin vascular responses to histamine introduced by iontophoresis, we designed a double-blind, randomized, two-period crossover trial in which 18 volunteers were randomized. METHODS: Before and 2, 5 and 7 h after drug administration, iontophoresis (30 s, 1.4 mA/cm2) of histamine 10% was performed and followed by (1) monitoring of skin vascular responses with LDF at the administration site and at 1 cm from it, and (2) wheal and flare area measurements. RESULTS: 2, 5 and 7 h after intake of the antihistaminic drug, there were significant differences between both drugs. Concerning LDF recordings, we noted at the histamine administration site an increase in perfusion unit (PU) values which is an effect known to be in proportion to the degree of inhibition of wheal reaction, and at 1 cm distal to the histamine administration site, there was a decrease in PU values. These changes were more marked under cetirizine. A greater suppressive effect of cetirizine on the wheal and flare reaction was consistently observed at all time points during the study, demonstrating its superior efficacy. CONCLUSION: We conclude that (1) cetirizine demonstrated a stronger antihistaminic effect compared to ebastine at all time points; (2) iontophoresis appears to be an appropriate method to study specific microvascular changes at the delivery site of histamine and hence to detect the earliest changes occurring at the site of agonist-antagonist competition in the skin. PMID- 9732165 TI - Application of argon plasma coagulation in skin surgery. AB - BACKGROUND: Argon plasma coagulation (APC) is a noncontact electrosurgical technique which has been used in open surgery for about 20 years and in endoscopy for about 5 years. OBJECTIVE: The aim of the present study was to provide preliminary results on the effect of this method in skin surgery. METHODS: The effects of variations in gas flow, output power and coagulation times on the extent of the resulting skin coagulation zone were tested in a pig skin model. Furthermore, preliminary results were obtained from 48 patients. RESULTS: The depth of the coagulation zone, which depends on the output power of the high frequency (HF) generator and the application time, was examined clinically and histologically in 150 different pig skin coagulations. The maximum depth of the coagulation zone was 4 mm. Coagulation zones below 4 mm can be maintained by controlling the output power of the HF generator and/or the duration of application. Variations of argon flow did not influence the depth of the coagulation zone significantly. In preliminary clinical tests, 48 patients with common warts, senile hemangiomas and actinic keratoses were treated with APC. In all cases, APC was highly effective and easy to perform. No severe problems or complications were observed. The skin lesions were destroyed with minimal or no scarring and without damaging the surrounding tissue. CONCLUSION: In the present preliminary study, APC proved to be an effective treatment with well-controlled tissue destruction. Further clinical studies are required to evaluate the suitability and the indications of this method in the treatment of cutaneous lesions. PMID- 9732166 TI - Treatment of vascular lesions using the VersaPulse variable pulse width frequency doubled neodymium:YAG laser. AB - BACKGROUND: The flashlamp-pumped dye laser is considered the treatment of choice for vascular lesions including port-wine stains. However, this treatment is associated with an intensive postoperative purpura and considerable pain. OBJECTIVE: We tested a newly developed laser device with a 532-nm neodymium:YAG laser with variable pulse widths between 2 and 10 ms. METHODS: Forty-two consecutive patients from our laser clinics were treated with the new laser system delivering energy through a chilled tip by means of circulating water (cooling temperature was between 4 and 5.5 degreesC). Forty of them had been pre treated with an argon laser and an argon-pumped dye laser. RESULTS: Out of the 42 patients, 1 patient had a complete clearing, 11 had an excellent result (more than 80% clearing), 21 patients had a good result (51-75% clearing), weak responses were seen in 9 patients. Significant success was already seen after 1 or 2 treatments. Side effects were rare. There was no purpura after treatment. Local anesthesia was only applied in 6 out of 42 patients. CONCLUSION: The variable pulse width frequency doubled neodymium:YAG laser seems to be an alternative to the flashlamp-pumped dye laser and deserves to be investigated in comparative trials. PMID- 9732167 TI - Relapses of onychomycosis after successful treatment with systemic antifungals: a three-year follow-up. AB - BACKGROUND: Data about relapses of onychomycosis after treatment with the new systemic antifungals vary among the different studies, with figures ranging from 3 to 20% for terbinafine and from 21 to 27% for itraconazole, depending on the follow-up duration. OBJECTIVE: To determine the prevalence of relapses of onychomycosis cured by terbinafine compared with that of onychomycosis cured by itraconazole. METHODS: We followed up 47 patients whose toenail onychomycosis had been mycologically cured in an open randomized study comparing intermittent itraconazole treatment with continuous terbinafine treatment and intermittent terbinafine therapy. Patients were examined every 3 months for up to 3 years after the end of therapy. At each visit clinical and mycologic (direct microscopy and cultures) evaluations were performed. RESULTS: Eight of the 36 patients (22.2%) who completed the study had a relapse of onychomycosis during the follow up period, including 2 patients of the terbinafine 250 mg group, 2 patients of the terbinafine 500 mg group and 4 patients in the itraconazole 400 mg group. As the original infection, the relapse was caused in all cases by Trichophyton rubrum. CONCLUSIONS: This study shows that 22.2% of patients with onychomycosis successfully treated with systemic antifungals experienced a relapse. The relapse rate increased from 8. 3% at month 12 to 19.4% at month 24 and to 22.2% at month 36. Relapses were more common in patients treated with pulse itraconazole (4/11) than in patients treated with continuous (2/12) or intermittent (2/13) terbinafine. Statistical analysis did not reveal any significant difference between relapse rates in the three groups. PMID- 9732168 TI - Combination of PUVAsol and topical calcipotriol in vitiligo. AB - BACKGROUND: A large variety of therapeutic agents are being tried for the treatment of vitiligo, but psoralens continue to be mainstay of treatment although they are not uniformly effective. Recent advances in pathophysiology have established a perturbed calcium homeostasis in affected skin, and melanocytes were shown to express vitamin D3 receptors. OBJECTIVE: The purpose of present study was to determine the efficacy of the combination of PUVAsol with topical calcipotriol in the treatment of vitiligo. METHODS: Nineteen patients with essentially bilateral symmetrical lesions were enrolled in a randomized, double-blind, right/left comparative study of 18 months duration. An oral dose of 0.6 mg/kg 8-methoxypsoralen was given 2 h before exposure to sunlight thrice weekly to all patients. The patients were advised to apply calcipotriol (50 microgram/g) on one side of the body and placebo ointment over the lesions on the other side twice daily. RESULTS: At the end of 6 months, 12 patients (70%) showed marked to complete improvement on calcipotriol-treated sides as compared to 6 patients (35%) showing similar improvement on placebo-treated sides (p <0.05). At the end of treatment, 13 patients (76%) showed marked improvement in calcipotriol treated lesions whereas 9 patients (53%) showed moderate to marked improvement in placebo-treated lesions. The repigmentation of hands and feet was much better with the combination of PUVAsol and calcipotriol. CONCLUSION: The combination of PUVA and calcipotriol is highly effective and works faster and may be used for shortening the therapy with PUVA in the treatment of vitiligo. PMID- 9732169 TI - Ecstasy pimples - a new facial dermatosis. AB - Ecstasy (XTC) has become a popular drug in the rave, dance and techno scene. Several severe disorders due to drug addiction have been described but no dermatological symptoms. We report on 2 patients (20-year-old female, 21-year-old male) with medical problems after taking XTC. Both developed a facial rash with reddish pimples after oral intake of XTC. The distribution resembled either periorificial dermatosis or acneiform rash without white- or blackheads. The lesions cleared without specific treatment. We suggest that XTC pimples represent an acneiform dermatosis in young people taking designer drugs. Though the dermatosis itself seems to be mild, it may be a cutaneous marker for drug abuse. PMID- 9732170 TI - Abscess-forming neutrophilic dermatosis: report of three cases associated with hemopathies. AB - BACKGROUND: The development of different types of neutrophilic dermatosis is reported to occur in the course of malignant hemopathies. These concern mainly Sweet's syndrome, pyoderma gangrenosum, erythema elevatum et diutinum and neutrophilic eccrine hidradenitis. OBSERVATIONS: We have recently encountered the cases of 3 patients who presented all with multiple acneiform papules and dome shaped aseptic abscesses leaving scars. Pus was sterile in all except case 3 in which slight Staphylococcus aureus growth was shown. However, in this patient, only steroids were effective demonstrating that this bacterium was not responsible for the disease. Histopathology disclosed a dense dermal polymorphonuclear neutrophil infiltrate and some mononuclear cells. Two of these patients had myelodysplastic syndromes while one had IgA myeloma. CONCLUSION: Abscess-forming neutrophilic dermatosis seems to be another type of neutrophilic dermatosis associated with hematological malignancies. PMID- 9732171 TI - HIV-Associated eosinophilic folliculitis and follicular mucinosis. AB - The term HIV-associated eosinophilic folliculitis (EF) designates an idiopathic dermatitis that appears in HIV-infected patients with different clinical manifestations but with a distinctive histological feature characterized by a predominantly eosinophilic infiltrate in the follicular infundibula. On the other side, follicular mucinosis (FM) is a reaction pattern in the follicular epithelium, characterized by a mucinous degeneration of the outer sheath of follicles and sebaceous glands. It has been described in association with a variety of unrelated conditions. We report 2 HIV-infected patients with a pruritic papular eruption. Histopathological study revealed the coexistence of EF and FM. To our knowledge, this is the first report that describes this association. The possible relationship between these two entities is discussed. PMID- 9732172 TI - Ethanol-induced urticaria: elevated tryptase levels after double-blind, placebo controlled challenge. AB - We present a 48-year-old patient who complained for 1 year about urticarial reactions which appeared always when he ingested alcoholic beverages. Skin prick tests with ethanol were negative but positive with 10% acetic acid in the patient. Normal controls tested negative with acetic acid. Skin prick tests to common immediate-type allergens were negative. The patient underwent a double blind, placebo-controlled challenge test. A few minutes after challenge with ethanol but not with placebo, the patient developed erythema and wheals on the chest and the upper arms. The tryptase serum level rose from undetectable (0.1 U/ml) before challenge to 3.8 U/ml after skin lesions had appeared. This case demonstrates that increased tryptase serum levels can help in the diagnosis of ethanol-induced urticaria. PMID- 9732173 TI - Photo Koebner phenomenon in erythema-multiforme-like eruption induced by contact dermatitis due to bufexamac. AB - A photo Koebner phenomenon following contact dermatitis due to bufexamac is described. A 52-year-old man presented with dermatitis on his buttocks and an erythema-multiforme (EM)-like eruption near the original lesion and on sun exposed areas. He had been using bufexamac ointment (Anderm(R)) around and inside the anus for a few weeks before he noticed exacerbation. On patch testing, he exhibited severe contact sensitivity to bufexamac. Phototesting performed on his first visit showed that he had an abnormal reaction to UVB. An EM-like eruption similar to the original lesions developed around the irradiated sites. In this case, photosensitivity does not seem to be a drug-induced photoallergy due to systemic or topical use, but rather a photo Koebner phenomenon. PMID- 9732175 TI - 80th annual meeting of the Swiss society for dermatology and venereology. PMID- 9732174 TI - Prevalences of subacute cutaneous lupus erythematosus and Epidermolysis bullosa acquisita among Korean/Oriental populations. PMID- 9732176 TI - Dr. C. Richard fleming. 1943-1997 PMID- 9732177 TI - Esophageal manometry and modern medicine. PMID- 9732178 TI - Helicobacter infection in man: problems to be solved. AB - Helicobacter pylori (Hp) infection in man has several disease outcomes, varying from asymptomatic chronic gastric inflammation to Hp-associated dyspepsia, pepic ulcer disease, gastric adenocarcinoma, and Malt lymphoma. Particularly controversial is the role of Hp infection in the genesis of chronic dyspeptic symptoms. Only a small percentage of chronic dispeptics have long-lasting remission of the complaints after cure of the infection. It is now well established that healing of the inflammation through microbial eradication cures peptic ulcer disease. The high efficacy of bismuth or PPI triple and quadruple therapies is overshadowed by the rising resistance to metronidazole and clarithromycin. The exact role of Hp in gastric carcinogenesis in the various geographical areas needs further study. The results of ongoing trials, evaluating the long-term outcome of Hp cure, on cancer rates are anxiously awaited. The acquisition rate of new knowledge through basal and clinical Hp research has rarely been witnessed in medicine. PMID- 9732179 TI - Clinical value of esophageal motility testing. AB - Esophageal motility testing is the method of choice in evaluating esophageal motor disorders. Some physicians, however, question the clinical utility of esophageal motility testing, since the results are often normal in symptomatic patients. The clinical utility of esophageal motility testing is reviewed for patients with a complaint of noncardiac chest pain, dysphagia or symptoms of gastroesophageal reflux disease. Esophageal motility testing is particularly useful for evaluating patients with dysphagia, but less so for gastroesophageal reflux disease patients, and has little clinical utility in patients with noncardiac chest pain. PMID- 9732180 TI - Hepatobiliary diseases in patients with AIDS: focus on AIDS cholangiopathy and gallbladder disease. AB - Diseases of the biliary tree and gallbladder are being described with increasing frequency among patients with the acquired immunodeficiency syndrome (AIDS). Despite the profound immunosuppressive therapy transplant recipients receive, these patients do not appear to be affected by these disorders; thus, human immunodeficiency virus (HIV)-infected patients appear to be uniquely susceptible. Most HIV-infected patients that develop cholangiopathy have severe immunodeficiency with CD4 lymphocyte counts <200/mm3. The objective of this review is to summarize the available literature on AIDS-cholangiopathy, focusing on its diagnosis and classification and to suggest an approach for its evaluation and its management. PMID- 9732181 TI - Somatostatin in the treatment of non-variceal upper gastrointestinal bleeding. AB - The efficacies of somatostatin and octreotide have been widely studied in the control of bleeding from oesophageal varices. It has also been suggested that these drugs may be useful for the control of non-variceal upper gastrointestinal (UGI) bleeding, including that from peptic ulcers. In approximately 80% of patients presenting with non-variceal UGI bleeding, haemorrhage ceases spontaneously and does not recur. However, the remaining 20% of patients require active treatment. Results from recent studies have indicated that somatostatin is an effective treatment for the control of non-variceal UGI bleeding in high-risk patients, i.e. those in whom haemorrhage does not cease spontaneously or is likely to recur. In contrast there is no good evidence available at present to support a role for octreotide in this indication. The efficacy of somatostatin in controlling bleeding in patients with non-variceal UGI bleeding at high risk of mortality upon admission, or rebleeding following endoscopy, coupled with an excellent safety and tolerability profile, suggests that it may be a valuable therapeutic option in the management of non-variceal bleeding. PMID- 9732182 TI - Risks of intra-abdominal nonshunt surgery in cirrhotics. AB - We have reviewed the risks of various nonshunt intra-abdominal operations in cirrhotic patients. Most of these studies are retrospective reviews with limitations. Among various risk stratifications in cirrhosis, Child-Pugh classification is sufficiently informative. Elective surgery can be done safely in patients with Child's A or B class. Operations in Child's C patients and emergent surgery carry formidably high mortality. Limiting the extent of surgery, controlling ascites, correcting coagulation abnormality and malnutrition and aggressively treating infection, might reduce mortality. Laparoscopic cholecystectomy and endoscopic sphincterotomy in cirrhotics seem to be promising in reducing mortality and morbidity. PMID- 9732183 TI - Extraesophageal varices. AB - Esophageal varices are the most common site of variceal bleeding. However, bleeding from varices that are not located in the esophagus may account for up to 30% of all variceal bleeding. Significant varices can occur in the stomach, duodenum, jejunum, ileum, colon, rectum, and biliary tree. They can also occur at the site of a surgical ostomy. These types of varices bleed less commonly than esophageal varices, but they can also be far more difficult to diagnose and treat. The absence of stigmata of recent esophageal variceal bleeding and certain clues in the patient's history and clinical presentation should raise the clinician's suspicion of an extraesophageal site of variceal bleeding. Particularly, patients with extrahepatic causes of portal hypertension, cirrhotic patients with a prior history of gastrointestinal surgery, and patients who present with profound bleeding but without hematemesis need to be evaluated further if an obvious site of esophageal variceal bleeding is not seen on initial endoscopy. In this article, we review the features particular to extraesophageal varices as well as the diagnosis and management of bleeding from these varices. We use the term extraesophageal rather than ectopic because the term ectopic varices implies exclusion of both esophageal and gastric varices. PMID- 9732184 TI - Imaging techniques in the evaluation of adenocarcinoma of the pancreas. AB - Adenocarcinoma of the pancreas is a highly malignant neoplasm that presents late and carries a dismal prognosis. Despite technical advances, the various imaging modalities used to evaluate and stage a pancreatic adenocarcinoma have not had a significant impact on survival rates. This article describes the various imaging modalities used to image and stage an adenocarcinoma of the pancreas and reviews their relative accuracy and limitations in diagnosing and staging this carcinoma. We conclude that for appropriate staging of adenocarcinoma of the pancreas, the combination of a computed axial tomography scan and an endoscopic ultrasound represents an efficient and cost-effective approach. PMID- 9732185 TI - Pathophysiology and therapy of chronic radiation-induced injury to the colon. AB - Radiotherapy of pelvic malignancies causes chronic radiation damage to the gut in approximately 5% of patients. The injury can lead to local ischemia and fibrosis with the development of ulcers, strictures and lower gastrointestinal bleeding. The clinical presentation varies from mild disease to debilitating rectal bleeding, diarrhea, obstruction and fistula formation. Therapy should be directed toward the dominant symptom. Formalin instillation and endoscopic obliterative therapy can be used for bleeding due to telangiectasis. Nutritional intervention, e.g. total parenteral nutrition or elemental diets, is useful as adjunctive therapy to maintain hydration and nutritional status. Surgery should be reserved for severe refractory bleeding, fistulas or obstruction. Due to the recurrent character of the disease and the high complication rate, surgery should be viewed as an effort of last resort. PMID- 9732186 TI - History of oral mucosa. PMID- 9732187 TI - Sustained-release alfuzosin, finasteride and the combination of both in the treatment of benign prostatic hyperplasia. European ALFIN Study Group. AB - OBJECTIVES: To assess the additive benefit of combining an alpha1-blocker and a 5alpha-reductase inhibitor. METHODS: This European, randomized, double-blind, multicenter trial involved 1.051 patients with lower urinary tract symptoms related to benign prostatic hyperplasia. Patients received sustained release (SR) alfuzosin (n = 358), a selective alpha1-blocker given at a dose of 5 mg twice daily without dose titration; finasteride (n = 344), 5 mg once daily, or both drugs (n = 349), for 6 months. Primary efficacy criteria were symptomatic improvement (International Prostate Symptom Score: I-PSS) and maximum flow rate (Qmax). Safety was assessed by monitoring adverse events. RESULTS: Symptomatic improvement was significantly higher from the 1st month of treatment with SR alfuzosin, alone or in combination; mean changes in I-PSS versus baseline at end point were -6.3 and -6.1, respectively, compared with -5.2 with finasteride alone (SR alfuzosin vs. finasteride, p = 0.01; combination vs. finasteride, p = 0.03). The percentages of patients with a decrease in I-PSS of at least 50% were 43, 42 and 33% for SR alfuzosin, the combination and finasteride, respectively (SR alfuzosin vs. finasteride, p = 0.008; combination vs. finasteride, p = 0.009). In the overall population, increases in Qmax were greater with SR alfuzosin and the combination, compared with finasteride alone after 1 month of therapy, but changes at end-point were similar in the three treatment groups. In those 47% of patients likely to be obstructed (baseline Qmax <10 ml/s), however, mean increases in Qmax were significantly higher with SR alfuzosin, alone or in combination, whatever the visit. Finasteride, alone or in combination, significantly impaired sexual function. The incidence of postural symptoms was low and similar in the three treatment groups. CONCLUSION: In this 6-month trial, SR alfuzosin was more effective than finasteride, with no additional benefit in combining both drugs. PMID- 9732188 TI - Ratio of free to total prostate-specific antigen in patients with prostatic intraepithelial neoplasia. AB - OBJECTIVE: There are many reports about the effects of prostatic intraepithelial neoplasia (PIN) on serum prostate-specific antigen (PSA) level. The aim of this study was to determine the relationship between PIN and serum free PSA/total PSA (fPSA/tPSA) ratios. METHODS: We evaluated 46 patients with PIN, 15 patients with benign prostatic hyperplasia (BPH), and 16 patients with localized prostatic carcinoma (CaP) for the amount of fPSA and tPSA with the chemiluminescent enzyme assay. RESULTS: fPSA values from BPH to high-grade PIN (PIN2 and PIN3) was increased, and then a decrease was observed from high-grade PIN to CaP. fPSA was significantly different between BPH and low-grade PIN and high-grade PIN. There was no significant difference observed between BPH and CaP. tPSA values increased from BPH to CaP. tPSA was significantly different between BPH and high-grade PIN and CaP. fPSA/tPSA ratios decreased from BPH to CaP. This ratio was significantly different between CaP and BPH and low-grade PIN. There was no significant difference between CaP and high-grade PIN. CONCLUSIONS: Our results confirm that fPSA/tPSA ratio is better at discriminating between patients with CaP and those with BPH, but not between patients with CaP and those with high-grade PIN. Due to similarities between CaP and high-grade PIN, we think that decreased fPSA/tPSA ratio obtained at the time of intial diagnosis of PIN without concurrent carcinoma could be used as predictive factors to distinguish patients in whom carcinoma will be found on subsequent biopsies from those with PIN not associated with cancer on repeat biopsy. PMID- 9732189 TI - Cryoablation of localized prostate cancer. Experience in 48 cases, PSA and biopsy results. AB - OBJECTIVES: As the first German center to perform perineal cryoablation of localized prostate cancer, we present our experience in a series of 48 consecutive patients. METHODS: 7 patients staged T1, 21 with T2 disease and 20 patients with T3 tumor were treated. 62.5% of the patients received neoadjuvant hormonal downsizing. Follow-up ranged from 4 to 27 months with a median of 15 +/- 5.7 months. RESULTS: Positive control biopsies after 6 months were obtained in 0% of T1 tumors, 16.7% of T2 tumors and 26.7% of T3 tumors. Prostate-specific antigen persistence above 1 ng/ml was diagnosed in 14.3, 33.3, and 40%, respectively. Complications were acceptable. 22.9% of the patients had prolonged urinary retention, requiring transurethral resection in 5 patients (10.4%) to relieve obstruction. In 5 cases (10.4%) incontinence was found, in 2 of these patients mild urge incontinence declined over time, in 3 cases moderate to severe stress incontinence developed. Two of these patients were pretreated with radiotherapy. No fistulae were noted. CONCLUSIONS: Cryoablation of the prostate is not a substitution for radical prostatectomy but enables the surgeon to perform a radical curative procedure in patients unfit for other radical forms of treatment or unwilling to undergo these. Long-term follow-up and prospective studies are necessary to define the clinical significance of this procedure. PMID- 9732190 TI - Two-year follow-up of a prospective randomised trial of electrovaporization versus resection of prostate. AB - OBJECTIVES: Transurethral electrovaporization of the prostate (TUVP) has become a popular, minimally invasive procedure to treat BPH with promising initial results. This study was conducted to compare the efficacy, safety and durability of TUVP with standard TURP. We report the 2-year follow-up. METHODS: 104 consecutive men with BPH admitted for surgery were randomised to TUVP (52 patients, mean age: 67.5 years) or TURP (52 patients, mean age: 70.2 years). 47 patients in each arm completed 2-year follow-up. RESULTS: Follow-up data at 2 years show a comparable, significant and maintained improvement in mean IPSS (TUVP: 4.3 vs. TURP: 6.3), quality of life score (TUVP: 1. 1 vs. TURP: 1.7), and maximum flow rate (TUVP: 22.4 vs. TURP: 21.2 ml/s) with fall in mean post-void residual volume (TUVP: 18.8 vs. TURP: 22.8 ml). Postoperative impotence reported in TUVP: 17% vs. TURP: 11% (p = 0.49) and retrograde ejaculation TUVP: 72% vs. TURP: 89% (p = 0.47). Two patients in each arm (4%) had urethral stricture and 2 patients (4%) in the resected group had bladder neck stricture. Four patients in each group required re-operation for residual adenoma during the 2 years (4% in each arm each year). CONCLUSIONS: Our 2 years' follow-up results suggest that TUVP is as effective as standard TURP in the treatment of moderate-sized BPH with comparable durability. PMID- 9732191 TI - Effect of laser prostatectomy on the serum prostate-specific antigen concentration: results of a prospective study. AB - OBJECTIVE: To assess the effect of the laser prostatectomy (LP) procedure on the serum prostate-specific antigen (PSA) levels. PATIENT AND METHODS: The serum PSA level was determined in 41 patients with benign prostatic hyperplasia 1 day before and 1, 3, 7, 15, 30, and 90 days after LP. All patients underwent preoperative evaluation with routine blood tests, serum PSA level, IPSS symptom questionnaire, intravenous pyelography, uroflowmetry, postvoid residual urine measurements, and transrectal ultrasonography (TRUS). IPSS symptom questionnaire, uroflowmetry, postvoid residual urine measurements, and TRUS were repeated 3 months after LP. RESULTS: PSA levels showed a statistically significant increase 24 h after LP, then a slow decrease and by 1 month the PSA levels had returned to their initial levels. A statistically significant positive correlation was found between the PSA level 24 h after LP and the amount of energy applied to the prostate during operation (r 0.87, p < 0.0001). After 30 and 90 days the mean PSA values were under the preprostatectomy concentration. The mean PSA values at 30 and 90 days were statistically significantly lower than those measured before treatment (p < 0.05). There was a statistically significant positive correlation between the reduction in PSA and the reduction in prostate weight 3 months after LP. CONCLUSION: This study showed that LP produced a variable rise in PSA, with a peak rise in PSA occurring 24 h after the procedure. In some patients, the serum PSA returned to baseline by 15 days. But, after 15 days the mean PSA level was still greater than the preprocedure value. Therefore, we recommend that blood should not be sampled for PSA for at least 30 days after LP. The mean PSA levels 30 and 90 days after treatment were lower compared with the preoperative levels. This low level of PSA can probably indicate a reduction in prostate volume following LP. PMID- 9732192 TI - Functional evaluation of different ileal neobladders and ureteral reimplantation techniques. AB - OBJECTIVES: To compare and assess the function of ileal neobladders with different reconfiguration and with several types of ureteral reimplantation. METHODS: Forty-five male patients underwent radical cystectomy and detubularized ileal neobladder. In 20 patients an ileal neobladder was carried out according to Studer's technique, in 12 a 'W'- and in 13 a 'U'-shaped neobladder. In the Studer's patients 60 cm of ileum was used, in the 'W' 40 and in the 'U' 30 cm. For the uretero-ileal anastomosis Nesbit's technique was utilized in the Studer's, in the 'W' and 'U' neobladders Camey Le Duc's technique was performed instead. Four patients underwent a serous-lined extramural tunnel reimplantation. Follow-up included a functional and morphological study of the urinary system and a urodynamic study. RESULTS: All Nesbit's uretero-ileum anastomoses resulted refluent when the reservoir was filled up, 15 of 50 ureteral reimplantations according to the Camey Le Duc technique showed reflux at full filling. At 3, 6 and 12 months follow-up, the double reconfiguration reservoirs (Studer's and 'W') showed a larger capacity and a lower maximum pressure than neobladders with a single bending. At 12 months, continence and the voiding interval time was significantly higher in the double reconfiguration than in the 'U' neobladders. CONCLUSION: The double reconfiguration of the reservoir ('W') might be preferable to that with a single one. As for the type of ureteral anastomosis to select, the problem is still debatable even if in our case-control study we have had better results in terms of reflux and stenosis with the uretero-enteric anastomosis with Nesbit's and associated afferent long tubular ileal limb than with Studer's technique. PMID- 9732193 TI - Extracorporeal shock wave lithotripsy for lower caliceal calculi. AB - OBJECTIVE: The aim of the study is to determine the effectiveness of extracorporeal shock wave lithotripsy (ESWL) therapy for isolated lower caliceal calculi. PATIENTS AND METHODS: We analyzed 165 patients who were treated with the Siemens Lithostar Plus on an outpatient basis between March 1993 and August 1997. The age of patients ranged from 17 to 70 (mean 39.11) years. The stone size varied from 4 to 42 mm, and patients who had stones larger than 21 mm had a double-J stent inserted prior to treatment. RESULTS: The overall stone-free rate at 3 months was 53.33%; whereas it was 61.79, 48.27, and 27.27% according to the stone size, /=21 mm, respectively. Complications were rare, including 2 pyelonephritis, 2 subcapsular hematoma formation, 24 renal colics and 8 stone streets, which were managed by ureteral stenting or additional ESWL and resulted in complete stone clearance. CONCLUSION: ESWL therapy is a reasonable and effective method for small lower caliceal stones, but due to its relatively low stone-free and high complication rates, percutaneous nephrolithotripsy or open surgery should be considered for stones larger than 21 mm. PMID- 9732194 TI - Requirement of analgesia for extracorporeal shock wave lithotripsy and efficacy of a nonsteroidal antiinflammatory drug: piroxicam. AB - OBJECTIVE: In this study, the requirement of analgesia and the analgesic efficacy of a long-acting nonsteroidal antiinflammatory drug (NSAID), piroxicam, were investigated in patients with renal stone disease treated with extracorporeal shock wave lithotripsy (ESWL). METHODS: This randomized, placebo-controlled study included 60 patients. Patients were divided into two groups randomly. A single dose of saline (2 ml) was given to the patients in group 1 (n = 20) and 2 ml of 40 mg piroxicam to the patients in group 2 (n = 40). All injections were administered into the gluteal muscle 45 min before ESWL. A verbal rating scale (VRS) was used to evaluate the pain. Groups were compared according to age, sex, weight, height, stone size, number of shock waves, duration of ESWL and VRS scores. RESULTS: There was no statistically significant difference between both groups in demographic data, stone size, number of shock waves and duration of ESWL procedure (p > 0.05). However mean VRS scores were significantly lower in group 2 than in group 1 during and after the ESWL procedure. CONCLUSION: We considered that analgesic agents should be used to control the pain in second generation lithotriptors. Piroxicam has clinically significant effects on the pain and also antiinflammatory effects, inhibits ureteric activity, and helps in stone passage. PMID- 9732195 TI - Cosmetic orchiectomy using pedicled fibro-fatty tissue graft for prostate cancer: a new approach. AB - OBJECTIVE: To preserve the appearance of a normal scrotum at orchiectomy, a pedicled fibro-fatty tissue graft was inserted in the lumen of the tunica vaginalis. METHODS: 42 testes of 21 patients with metastatic prostate cancer were treated. A pedicled fibro-fatty tissue graft prepared from the region between the spermatic cord and the scrotal septum was inserted in the lumen of the tunica vaginalis following epididymis-sparing orchiectomy under local or spinal anesthesia. RESULTS: A pedicled fibro-fatty tissue graft which reached the epididymal tail was prepared in all instances. The average postoperative volume of the scrotal content was 18 ml (range 5-30 ml). Postoperative complications were scrotal edema, sensation of traction at the inguinal region and epididymitis. All but 1 patient were satisfied with the appearance of the scrotum and achieved sufficient androgen ablation. Mean follow-up was 8 (3-11) months. CONCLUSION: This cosmetic orchiectomy is a safe, simple and economical procedure to preserve the appearance of a normal scrotum; it also achieves the reliable result of androgen deprivation. PMID- 9732196 TI - Using buccal mucosa for urethral reconstruction in primary and re-operative surgery. AB - OBJECTIVE: Urethral reconstruction using buccal mucosa grafts finds increased broader use in patients with congenital or acquired urethral defects. The authors present their experience of 20 such treated patients. 12 repeatedly operated patients (11 congenital defects, 1 war injury) showed urethral defects of 4.5-20 cm in length. In 8 patients, 6 with hypospadias and 2 with prior straightening of the penis due to penoscrotal transposition, a urethral defect length of 2.5-7 cm had to be bridged. METHOD: For 2 patients a tubularized buccal mucosa free graft was used and for 18 patients the onlay method was partly or wholly used. In 1 patient a tubularized preputial island flap was used in addition to a buccal mucosa free graft. The urethral meatus was placed at the tip of the glans in all patients. RESULTS: Postoperatively a coronary fistula occurred in 1 patient. In 2 patients anastomotic strictures were seen after 18 and 60 days, respectively. One patient had a meatal stenosis and one splitting of the glans wings. With a mean follow-up time of 27 months, no further complications have been observed. CONCLUSION: The use of the buccal mucosa graft for urethral reconstruction is a successful method with a low incidence of complications. The indications arising from this operating method should, in our opinion, lead to the increased and broader use of this treatment method for congenital and acquired urethral defects. PMID- 9732198 TI - Is resolution of vesicoureteric reflux by conservative management predictable in patients with myelodysplasia? AB - OBJECTIVE: To evaluate the probable factors that might predict the outcome of conservative management of vesicoureteric reflux (VUR) in myelodysplastic patients. PATIENTS AND METHODS: A retrospective review of 24 children with VUR secondary to neurogenic bladder (15 girls and 9 boys, age range 1-18 years) out of 75 myelodysplasia patients between 1994 and 1996 was made. Patients were grouped according to their response to conservative management: Group I: patients with their VUR resolved or downgraded (n = 15), and group II: patients with their VUR unchanged or increased (n = 9). The following parameters were compared between the two groups: age, sex, VUR grade and laterality, urodynamic parameters (bladder capacity, compliance, leak point pressure), type of bladder neuropathy, accompanying neuropathology (walking problem, anal incontinence). RESULTS: Most of the parameters studied failed to predict the outcome of conservative management of VUR in patients with neurogenic bladder dysfunction. Higher grades of VUR if bilateral seem to benefit more from conservative management than lower grades do. Conservative management appears to be more effective in hyperreflexic bladders than areflexic bladders in terms of VUR resolution. CONCLUSION: Although prediction of patients resistant to conservative management of VUR could save myelodysplastic children from prolonged risk of renal damage, current methods of evaluation are of very little help in this aspect. PMID- 9732197 TI - Clinical features of vesicoureteral reflux in infants and outcome of conservative therapeutic approach. AB - OBJECTIVES: We evaluated the clinical features of vesicoureteral reflux (VUR) detected in infants and the outcome of a conservative therapeutic approach. METHODS: Consecutively 67 infants with VUR (55 boys and 12 girls) were enrolled in this study. Of the 67 patients, 34 had primary and 33 had secondary VUR. Underlying abnormalities in secondary VUR were: posterior urethral valve (PUV) in 7; bulbar urethral stenosis (Cobb's collar) in 16; neurovesical dysfunction (NVD) in 8, and others in 2. Transurethral incision was performed in patients with PUV or Cobb's collar. NVD was managed with intermittent catheterization. All patients were followed with antibiotic prophylaxis. RESULTS: No significant difference was found in VUR grades between primary and secondary VUR. Spontaneous resolution of VUR was noted in 31% of primary and 54% of secondary VUR (p<0.02). VUR downgrading including VUR resolution was also noted more often in secondary than in primary VUR (80 vs. 48%; p<0.01). CONCLUSIONS: The distribution of primary and secondary VUR in infants was almost equal in our study. Resolution of reflux is seen more often in secondary than in primary cases. Thus, early detection and proper management of underlying lower urinary tract abnormalities, either structural or functional, are crucial in the treatment of VUR in infants. PMID- 9732199 TI - Persistent Mullerian duct syndrome. AB - Persistent mullerian duct syndrome (PMDS) is a rare form of male pseudohermaphroditism. We present 5 cases with PMDS (2 cases associated with testicular malignancy) and discuss the diagnosis and management. Management strategies of PMDS have changed. Whereas in the past, removal of the mullerian remnants was targeted together with orchidopexy or -ectomy, this is no longer recommended. However, testicles that cannot be descended at an early stage are at a high risk of malignancy and should, therefore, be removed. If this is necessary on both sides, there is the additional problem of lifelong testosterone substitution which requires efficient patient monitoring and good patient compliance. In cases where this cannot be achieved, compromises, such as temporarily delayed orchidectomy, may be considered. PMID- 9732200 TI - Absence of brain parenchymal damage following intravascular injection of polytetrafluoroethylene paste. AB - OBJECTIVE: Polytetrafluoroethylene (PTFE) paste has been used for over 30 years to treat urinary incontinence and for 14 years to treat vesicoureteral reflux with little reported morbidity. Some investigators have been concerned by the use of PTFE as the implanted substance, because migration of PTFE particles from the site of injection in the periurethral and periureteral regions to the lungs and brain has been reported in animal studies. We injected PTFE paste intravascularly in dogs in order to investigate its effect on brain parenchyma. METHODS: A total of 12 mongrel dogs, weighing 11.5-17.5 kg, were divided into four groups. Group 1 (n = 3): injection of 0.5 ml of PTFE paste suspended in 50 ml of saline into a peripheral vein once a week for 4 weeks. Group 2 (n = 3): injection of 50 ml of saline into a peripheral vein once a week for 4 weeks as controls. Group 3 (n = 3): one injection of 0.1 ml of PTFE paste suspended in 20 ml of saline into the right carotid artery. Group 4 (n = 3): one injection of 20 ml of saline into the right carotid artery as controls. After an interval of 6 months, all animals were sacrificed and the lungs and brain removed. Brain from 1 animal in each group was dissolved in sodium hypochlorite solution, the resulting organ suspension was centrifuged, and the smear preparations of the precipitate examined by polarized light microscopy, scanning electron microscopy, and X-ray microanalysis. Brains from 2 animals in each group were fixed in formalin solutions, 6-micrometer sections were cut and stained with haematoxylin and eosin, Cajal stain for Purkinje fibre, and Luxol fast blue stain for myelin, and glial fibrillary acidic protein immunohistochemistry was carried for astrocytes using monoclonal mouse anti-GFAP (glial fibrillary acidic protein) at a dilution of 1:50 with an avidin biotin-peroxidase complex method. RESULTS: PTFE particles were seen in the cerebral vessels in only those animals who had PTFE injected into the right carotid artery. The haematoxylin and eosin staining showed PTFE particles in vessels with focal foreign-body reaction, but no infarction. Luxol fast blue staining showed no demyelination around vessels containing the particles and the parenchyma. Cajal staining demonstrated no abnormality of nerve fibres, and there was no astrocytosis using GFAP immunohistochemical staining. CONCLUSIONS: Our findings indicate that following intravenous injection, there was no evidence of migration of PTFE to the brain. Small quantities of PTFE injected into the carotid arteries were associated with local foreign-body reaction, but no brain parenchymal tissue damage was found. PMID- 9732202 TI - 13th Congress of the European Society for Urological Oncology and Endocrinology. Innsbruck, Austria, October 1-3, 1998. PMID- 9732201 TI - Expression of p21(waf1/cip1) protein in transitional cell bladder tumours and its prognostic value. AB - OBJECTIVES: p21(waf1/cip1) protein is a cyclin-dependent kinase inhibitor able to arrest the cell at the G1 phase by inhibiting DNA replication through interaction with proliferating cell nuclear antigen (PCNA). Experimental analyses of human bladder cancer cell lines show that p21 might be considered a tumour suppressor gene since it is able to induce apoptosis like p53. The prognostic value and expression of p21(waf1/cip1) is incompletely studied in bladder cancer at present. METHODS: The expression of p21 protein was immunohistochemically analysed in paraffin-embedded specimens of 186 patients with primary transitional cell bladder tumours. The results of immunohistochemical analysis were related to known prognostic factors and complete long clinical follow-up data (over 11 years). RESULTS: The expression of p21(waf1/cip1) protein was significantly related to DNA ploidy, S phase fraction, mitotic index, apoptotic index, morphometric nuclear factors, and the expression of p53 and PCNA proteins, whereas it was unrelated to grade or TNM classification. In univariate survival analysis, the expression of p21(waf1/cip1) protein was not significantly related to prognosis. Independent prognostic factors were T category, papillary status and mitotic index. CONCLUSION: The results indicate that although the expression of p21(waf1/cip1) protein is related to indicators of cell proliferation, apoptosis and p53 expression, it has no better prognostic value than already established prognostic factors in bladder cancer. PMID- 9732203 TI - Fixation stability among schizophrenia patients. AB - Twenty-four schizophrenia and 26 normal subjects were presented targets for fixation at +/-17.5 degrees and 0 degrees of visual angle. The manner in which stimuli were presented allowed us to evaluate for the presence of gaze-evoked and rebound drifts, and for frequency of saccades at both eccentric and central fixation. Schizophrenia patients and normal subjects had remarkably similar performance regardless of stimulus condition. These results suggest that the gaze holding apparatus is functioning normally among schizophrenia patients, a finding that is not easily attributable to either medication effects and/or low statistical power. These data are another indication that schizophrenia patients have specific, not general, abnormalities of ocular motor control. PMID- 9732204 TI - Aberrant functional organization in schizophrenia: analysis of EEG coherence during rest and photic stimulation in drug-naive patients. AB - EEG coherence provides a measure of functional correlations between two EEG signals. The present study was conducted to examine intrahemispheric EEG coherence at rest and during photic stimulation (PS) in 18 drug-naive patients with paranoid schizophrenia and 30 control subjects. Compared with the controls, the schizophrenic patients had significantly higher intrahemispheric coherence of the resting EEG for the delta band, although no significant group differences were found for other frequency bands. EEG analysis during PS showed that the patients also had significantly higher EEG coherence over the left posterior regions. In this study, we also examined the changes in intrahemispheric coherence from rest to the stimulus condition (i.e., PS-related coherence reactivity); the patients were found to show significantly smaller changes, with significant group differences being also confined to the posterior regions in the left hemisphere. These findings provide evidence that schizophrenic patients have abnormal EEG coherence in both resting and stimulus conditions and suggest more diffuse, undifferentiated functional organization within hemispheres. In addition, diminished coherence reactivity suggests a failure of PS-related functional reorganization in schizophrenia. PMID- 9732205 TI - Differences in whole blood serotonin levels based on a typology of parasuicide. AB - Suicidal behavior has to be considered as a multifactorial phenomenon, which can be analyzed in a classifying-phenomenological manner. We have examined the relation of parasuicide typology to whole blood concentrations of serotonin, HVA, and tryptophan in 58 patients classified into 4 groups of parasuicide typology compared to 22 nonsuicidal depressed patients and 20 healthy subjects. Suicidal patients classified as impetuous, desperate and ambivalent types had significantly reduced whole blood 5-HT levels in comparison with the appealing type as well as nonsuicidal subjects. No differences were detected in the HVA content, but whole blood tryptophan concentrations were significantly reduced in impetuous suicidal patients and depressed patients compared to healthy subjects. This study provides evidence for reduced whole blood serotonin content based on different types of parasuicide. PMID- 9732206 TI - Relationship between cognitive function, growth hormone and insulin-like growth factor I plasma levels in aged subjects. AB - Basal growth hormone (GH) and insulin-like growth factor I (IGF-I) as well as GH responses to GH-releasing hormone (GHRH) were studied in 22 subjects (7 females, 15 males), aged between 65 and 86 years. The study was aimed at investigating the possible correlations between the age-dependent GH-IGF-I axis decline and the cognitive function - assessed by the Mini Mental State Examination (MMSE). The relationship between hormonal data, cognition and age, body weight, body mass index (BMI), some nutritional indices (triceps skinfolds, TSF, mid-arm circumference, MAC), and physical activity - quantified by the physical functioning index (PFI)--were also analyzed. GH basal levels were within the normal range, while GH responses to GHRH were blunted in most cases. GH peaks after GHRH were directly correlated with GH basal values. IGF-I serum levels were found to be in the lower part of the reference range for adult subjects or below it. GH responses to GHRH, but not GH and IGF-I basal levels, were inversely correlated with subject age. GH secretion areas after GHRH were inversely correlated with BMI, but no further correlations between GH data and clinical or nutritional parameters were found. MMSE values directly correlated with MAC and PFI values. IGF-I levels were directly correlated with MMSE scores, being lowered in patients with more advanced cognitive deterioration, and with MAC values--the decrease of which is thought to reflect protein caloric malnutrition--but not with body weight, BMI, TSF and PFI. MMSE-related protein caloric malnutrition and decreased physical activity possibly take part in affecting IGF- I function in subjects with mild cognitive impairment and, reciprocally, IGF-I decrement might affect neuronal function. PMID- 9732207 TI - Enlarged cerebrospinal fluid spaces in opiate-dependent male patients: a stereological CT study. AB - Computed tomography was performed in 9 male patients with a diagnosis of opiate dependence and in 9 age-matched psychiatric controls (neurotic depression). Patients with a history or diagnosis of another substance dependence (alcohol, cocaine, cannabis) were excluded from the study. The volumes of internal and external components of cerebrospinal fluid (CSF) were measured with a point counting stereological method. Analysis of variance with age as a covariate revealed a significant enlargement of external and external CSF spaces in male patients with opiate dependence. There was no significant correlation between the length of opiate dependence and the volumes of internal and external CSF spaces. The present results suggest that opiate dependence is associated with structural brain alterations. However, the relationship between opiate dependence and structural brain changes is complex and still not well understood. PMID- 9732208 TI - Upregulation of the platelet Serotonin2A receptor and low blood serotonin in suicidal psychiatric patients. AB - Suicidality has been found to be associated with low pre- and postsynaptic serotonin functioning. The purpose of this study was to examine whether in acutely suicidal psychiatric inpatients, the blood serotonin concentration was related to the underlying psychiatric disorder and whether it was associated with changes in the affinity (dissociation constant, KD) or in the maximal binding capacity (Bmax) of the platelet serotonin2A receptor. We therefore determined the blood serotonin concentrations and the platelet serotonin2A receptor activities of 45 suicidal psychiatric patients and 20 healthy subjects. We found that the blood serotonin concentrations were significantly lower in suicidal patients compared to healthy subjects. In all diagnostic categories (affective disorder, schizophrenia and adjustment disorder) we noted a significantly higher maximal binding capacity of the platelet serotonin2A receptor. These findings support the notion that a reduction in the availability of serotonin and an upregulation of the serotonin2A receptors in psychiatric patients are associated with a loss of control over suicidal impulses. PMID- 9732209 TI - Lymphocyte response to mitogens: influence of life events and personality. AB - The aim of the present study was to examine the possibility that the accumulation of life events is associated with low lymphoproliferative response to mitogens in undergraduate students. We also analyzed the possible interaction between life events and personality traits. Lymphocyte response to phytohemagglutinin (PHA) was lower in subjects with high life events compared to those with low levels. Introverted subjects were found to exhibit lower lymphocyte responses to PHA than those who were extraverted, and there was no interaction between the effect of introversion and life events on the proliferative capacity. Lymphocyte proliferation was low in subjects with high anxiety scores, whether they had high or low levels of life events. In the group with high scores on independence a high accumulation of life events was not associated with lower lymphoproliferation; while in the group with low scores it was, suggesting that independence buffers the association between life stress and lower cellular immunity. PMID- 9732210 TI - The cholinergic basis of the smoking-induced EEG activation profile. AB - Acute quantitative electroencephalographic effects of cigarette smoking were examined in 15 smokers within a repeated-measures design which assessed changes in power-spectral estimates following acute pre-treatment with placebo, a dose (20 mg) of mecamylamine, a dose (0.6 mg) of scopolamine and a combined dose of mecamylamine and scopolamine. Compared to sham smoking, the smoking of a single cigarette following placebo pre-treatment reduced absolute and relative power in slow (delta, theta) frequency bands, increased absolute and relative power in alpha and beta frequency bands and accelerated mean frequency. These smoking induced power changes in slow- and fast-frequency bands were differentially affected by the separate and combined actions of the cholinergic antagonists with treatments involving mecamylamine tending to abolish smoking-induced slow frequency absolute power reductions and fast-frequency relative power increments. Self-ratings of smoking-induced increases in alertness were altered by mecamylamine and combined treatments while sensory aspects of cigarette smoking were only altered with combined mecamylamine and scopolamine pre-treatment. The results are discussed with respect to brain-behaviour relationships and mechanisms maintaining the smoking habit. PMID- 9732211 TI - The statistical distribution of wrist movements during sleep. AB - The purpose of this work was to describe the basic statistical properties of the process of production of movements measured with a wrist actimeter, along a complete sleep period in normal human subjects. Two distinct types of random magnitudes were considered to analyze the data, the times between successive groups of movements and the number of movements at each fixed time (1 min) measurement epoch. Suitable probabilistic models for the two variates were chosen, fitting theoretical distribution functions to the observed data. It is concluded that interval data fit a one-parameter exponential distribution, while the number of movements fit a two-parameter negative binomial distribution. The estimated values of these parameters, besides being necessary to perform further statistical analysis, are a measure of the intensity and frequency of the movements. Finally the relationship between polysomnography measures and the elicited parameters was studied. PMID- 9732212 TI - Strontium-89 chloride in the treatment of bone metastases from breast cancer. AB - Sixty-four patients with painful metastatic breast cancer in bone were treated with 2 MBq/kg of strontium-89 chloride as a single intravenous injection. Patients were followed with records of medication, hematology parameters, serial bone and Sr-89 bremsstrahlung images and with a point pain score scale (10-0). The response was assessed during a 6-month period of follow-up. Fifty-two of 64 patients (81%) showed at least a moderate improvement. Eighteen out of the 52 responders showed a dramatic decrease in bone pain (35%), 21 (40%) presented a satisfactory response and in 13 cases (25%) the response was moderate. Only 12 patients (19%) from the whole group did not feel any improvement on pain palliation. A statistically significant decrease of pretreatment levels of platelets and leukocyte counts was observed after 4-6 weeks of therapy in 50 (70%) patients. Although most patients showed no change in their bone scans after 3 months of treatment, an obvious improvement was observed in 3 of them. Furthermore no additional painful metastases on their bone scintigraphic images were observed. The selective strontium-89 local uptake in metastatic sites was also confirmed directly by bremsstrahlung scans which were absolutely comparable to the respective 99mTc bone scans. Precautions have been taken against Sr-89 contamination from the patients' blood or excretions. PMID- 9732213 TI - Proliferating cell nuclear antigen immunoreactivity and the presence of P53 mutation in basaloid squamous cell carcinoma of the larynx. AB - Basaloid squamous cell carcinoma (BSCC) is an aggressive variant of squamous cell carcinoma (SCC) that was first described in 1986. In the English literature, only 28 cases of this histopathological entity have been reported in the larynx. Proliferative features of this infrequent tumor were investigated by proliferating cell nuclear antigen immunoreactivity, and the presence of P53 mutation was also investigated in a series of 4 cases. BSCC was found to share similar features with regular SCC in terms of its proliferation rate and P53 mutation positivity, as well as biological behavior and clinical outcome. PMID- 9732214 TI - First-line vinorelbine-mitoxantrone combination in metastatic breast cancer patients relapsing after an adjuvant anthracycline regimen: results of a phase II study. AB - PURPOSE: Previous studies demonstrated that doxorubicin and vinorelbine combinations in first-line chemotherapy are highly active in metastatic breast cancer. Mitoxantrone is an anthracenedione with low cardiotoxicity, and seems to be effective when combined with vinorelbine after prior exposure to anthracyclines. PATIENTS AND METHODS: Seventy-two patients with metastatic breast cancer were included in a phase II study. All patients had previously received one anthracycline-containing regimen (doxorubicin or epirubicin) in an adjuvant setting. Vinorelbine was administered at 25 mg/m2 in a 20-min intravenous (i. v.) infusion, days 1 and 8. Mitoxantrone was given at 10 mg/m2 (66 patients) or 12 mg/m2 (6 patients) in a slow i.v. infusion on day 1. Courses were repeated every 3 weeks. RESULTS: Sixty-five patients were evaluable for response; the objective response rate was 49% (95% CI: 37-63%), including four complete and 28 partial responses, with a median duration of response of 7 months (range 2.3-27). Median overall survival was 19 months (range 2-48). Grade 3-4 granulocytopenia was observed in 46% of patients. There were 12 admissions (3% of cycles), involving 17% of patients for febrile neutropenia. Seven patients (10%) experienced grade 3 or 4 cardiotoxicity, and 1 patient died of cardiac heart failure. Other side effects were rare and mild. CONCLUSIONS: The vinorelbine and mitoxantrone combination is an active regimen with low toxic complications when cumulative doses of mitoxantrone are limited to 70 mg/m2. The results in this phase II study make it worthwhile including this regimen in a phase III study for patients who have previously received an anthracycline-containing regimen. PMID- 9732215 TI - A case control study on risk factors involved in inflammatory breast recurrence after breast-conserving surgery. AB - Recurrence that poses the biggest problem after breast-conserving surgery is local recurrence. Particularly, in the case of inflammatory breast recurrence which is rare but has a specific pathologic nature, it is important to elucidate the pathology and risk factors and to consider appropriate countermeasures. In the present study, we classified 133 cases of recurrence following breast conserving surgery, collected from 18 key hospitals/institutes in Japan. Recurrence types were divided into three groups, namely, inflammatory breast recurrence, noninflammatory breast recurrence and distant metastasis only, and the risk factors involved in recurrence were investigated by the case control study allotting 2 controls to each case. The study population consisted of 9 cases of the inflammatory type, 64 cases of the noninflammatory type and 60 cases of distant metastasis. The significant risk factor for inflammatory breast recurrence was positive lymph node metastasis, which was significantly more frequent in lymphatic invasion-positive cases unlike in the distant metastasis group. The positive surgical margin and nonradiation therapy which have been shown to be significant risk factors for noninflammatory breast recurrence were entirely unrelated with inflammatory breast recurrence. In addition, the inflammatory-type recurrence time was as short as about 12 months irrespective of whether radiation therapy was performed or not. The inflammatory type was accompanied with local wide extension (cancerous embolus of the dermal lymphatic vessels), and distant metastasis (lymphangitis carcinomatosa) at the time of recurrence, and further surgery was impossible in most cases, with a significantly poorer prognosis than the other recurrence types. These findings suggest that this recurrence corresponds to the so-called 'occult' case of primary inflammatory breast carcinoma. We think it important to predict this recurrence by close pathological examination, particularly in patients with lymph node metastasis, and to consider appropriate measures. PMID- 9732216 TI - Clinical significance of combined immunohistochemical detection of CD44v and sialyl LeX expression for colorectal cancer patients undergoing curative resection. AB - To evaluate their prognostic value, the expressions of CD44v and sialyl LeX (SLX) in colorectal cancers were studied immunohistochemically. Tissue specimens were reacted with monoclonal antibodies (mAb) CD44-1V and CSLEX-1. Of the 145 colorectal cancer patients undergoing curative resection, 59 (40.7%) were positive for mAb CD44-1V, and 40 (27.6%) were positive for mAb CSLEX-1. There was a significant correlation between the combined expression of SLX and CD44v8-10 and lymph node metastasis. The patients with tumors negative for CD44v8-10 and SLX had the most favorable prognoses. Conversely, the patients with tumors positive for both CD44v8-10 and SLX had a high recurrence rate and the poorest prognoses. In a multivariate analysis using the Cox regression model, the combined expression of SLX and CD44v8-10 emerged as an independent prognostic indicator. These results suggested that the combined expression of CD44v8-10 and SLX may be a biologic marker of prognostic significance. PMID- 9732217 TI - Diffuse large cell lymphomas: identification of prognostic factors and validation of the International Non-Hodgkin's Lymphoma Prognostic Index. A Hellenic Cooperative Oncology Group Study. AB - Several clinical prognostic factors have been identified that predict treatment outcome in patients with diffuse large cell lymphomas. An International Non Hodgkin's Lymphoma Prognostic Index (IPI) has been recently formulated. We tried to identify the clinical prognostic factors that predict treatment outcome in Greek patients with diffuse large cell lymphomas and validated the IPI in these patients. The possible prognostic variables for tumor response, relapse-free (RFS) and overall survival (OS) were analyzed in 239 consecutive patients treated with anthracycline-based chemotherapy regimens. In univariate analysis, factors associated with poor response were stages III-IV, performance status (PS) >/=2, spleen and bone marrow involvement, more than one extranodal site involved, increased lactate dehydrogenase (LDH) value, hemoglobin (Hb) <12 g/dl, albumin <3.5 g/dl, erythrocyte sedimentation rate (ESR) >50 mm/h. Multivariate analysis identified stage, PS, more than one extranodal site involved, increased LDH level, and ESR > 50 mm/h as the factors more predictive of poor response. For RFS, multiple Cox analysis found stages III-IV and bone marrow involvement to be statistically significant. For OS, multiple Cox analysis identified stage III-IV, PS >/=2, bone marrow involvement, more than one extranodal site involved, increased LDH level and ESR > 50 mm/h as negative prognostic factors. Patients stratified in the different risk groups of the IPI had a significantly different outcome regarding complete response (CR) rate, RFS and OS. In conclusion, although age >60 years was not recognized as an adverse factor in this analysis, our patients stratified in the different groups of the IPI had significant differences in CR rate, 2-year RFS and OS verifying the prognostic significance of the index. Bone marrow involvement and ESR > 50 mm/h, parameters that are not included in the IPI, adversely affected survival. PMID- 9732218 TI - Fadrozole versus megestrol acetate: a double-blind randomised trial in advanced breast cancer. AB - Ninety-six patients were entered into a randomised, double-blind, double-dummy, clinical trial to assess the efficacy and safety of fadrozole as compared to megestrol acetate as second-line hormonal treatment for patients with advanced breast cancer. Analysis of results was on an intention-to-treat basis and included response rate, time to progression (TTP), time to treatment failure (TTF) and survival. Forty-six patients received fadrozole and 50 were randomised to megestrol acetate. Patients and pretreatment prognostic variables were balanced in the two arms of the trial. The objective response rates [3/46 (7%) for fadrozole and 3/50 (6%) for megestrol acetate], TTP, TTF and survival were similar in the two arms of the trial. Toxicity was also similar in the two arms of the trial and consisted mainly of oedema, hypertension and minor gastrointestinal symptoms. Fadrozole appears to be as active as megestrol acetate in second-line hormonal treatment of advanced breast cancer. PMID- 9732219 TI - Clinical evaluation of pancreatitis-associated protein as a serum marker of hepatocellular carcinoma: comparison with alpha-fetoprotein. AB - This study evaluated the significance of serum pancreatitis-associated protein (PAP) assay, as a marker of hepatocellular carcinoma (HCC), in comparison and combined with alpha-fetoprotein (AFP) assay. Sixty-five patients with HCC, 59 with liver cirrhosis (LC) and 68 asymptomatic controls (C) were studied. PAP and AFP values significantly increased from C to LC and HCC group (p < 0.0001). The area under receiver-operating characteristic (ROC) curve for the two markers was not statistically different. At 100% specificity, ROC analysis gave a cut-off level for AFP of 166 IU/l with 40% sensitivity, and a cut-off level of 240 microg/l for PAP with 23% sensitivity. Diagnostic accuracy of combined AFP and PAP assay was significantly higher than AFP alone. Sensitivity according to tumor size also improved using the combined assay, especially for tumors <5 cm. Stepwise logistic regression indicated that AFP, but not PAP, was associated with an increased risk of developing HCC. These data confirm that PAP production is increased only in some cases of HCC and that the combined PAP and AFP assays do not significantly improve specificity over AFP assay alone. Consequently, PAP assay can only be recommended in cases of justified suspicion of HCC with negative AFP. PMID- 9732221 TI - Rare pulmonary tumors - a review of 32 cases. AB - In a review of pulmonary tumors diagnosed at our institute from 1976 to 1995, we found 20 malignant and 12 benign rare tumors, which accounted for 0.57 and 0.34% of all pulmonary tumors, respectively. The histological types of these rare malignant tumors were malignant lymphoma (6/20), carcinosarcoma (3/20), mucoepidermoid carcinoma (2/20), bronchial gland mixed tumor (2/20), adenocystic carcinoma (1/20), myoepithelioma (1/20), leiomyosarcoma (1/20), epitheloid hemangioendothelioma (1/20), hemangiopericytoma (1/20), malignant melanoma (1/20) and choriocarcinoma (1/20). Benign rare tumors involved papilloma (3/12), lipoma (3/12), leiomyoma (3/12), adenoma (1/12), fibroma (1/12), and meningioma (1/12). The clinical and pathological features of malignant tumors were roughly the same as those of common pulmonary carcinomas. In contrast, benign tumors were never larger than 3 cm and were more commonly located in the central parts of the lung, which explained the relatively frequent symptoms of wheezing and fever associated with obstructive pneumonia. PMID- 9732220 TI - Combination regimen with carboplatin, epirubicin and etoposide in metastatic carcinomas of unknown primary site: A Hellenic Co-Operative Oncology Group Phase II Study. AB - The encouraging results that have been reported for cisplatin combination chemotherapy in a minority of patients with cancer of unknown primary (CUP) together with the previously shown equal activity of carboplatin in this setting, prompted us to investigate the effectiveness of a carboplatin-containing regimen in a phase II clinical trial. Sixty-two evaluable CUP patients entered the protocol. The chemotherapy regimen consisted of carboplatin 300 mg/m2 IV D1, epirubicin 45 mg/m2 IV D1 and etoposide 120 mg/m2 IV D1-3 (CEE regimen) administered every 3 weeks. The median age of the patients was 61 years and 36 were male. Thirty-one diagnosed as poorly differentiated carcinomas (pdc) and the rest as adenocarcinomas. By clinicopathological criteria, 14 patients had a predominately nodal disease with pdc or poorly differentiated adenocarcinomas (pda), 3 women peritoneal carcinomatosis, and the remaining 46 patients had a predominately splanchnic involvement (24 pdc, 22 pda). Twenty-three patients responded to chemotherapy with 4 (6.5%) complete and 19 (30.5%) partial responders (RR 37%, 95% CI 25-49%). An equal activity of the regimen was observed between the two major histopathological types, the pdc and the adenocarcinomas. Nevertheless, significant differences were seen when the CEE regimen was assessed for its activity in the distinct clinicopathological subsets of CUP. Patients with predominately nodal disease of midline distribution with pdc or pda, and women with peritoneal carcinomatosis, achieved a response rate of 64 and 62% respectively, as compared with a 26% response rate for those with predominately splanchnic involvement. Overall median survival was 10 months and for patients with midline distribution 15 months. The regimen was well tolerated. It is concluded that CEE is a relatively nontoxic chemotherapy regimen and easily administered on an outpatient basis. This prospective phase II study confirmed the activity of carboplatin in the chemosensitive subsets of the predominately nodal disease of midline distribution and peritoneal carcinomatosis in women in the CUP syndrome. PMID- 9732222 TI - Monoclonal antibody targeting of ovarian carcinoma. AB - The ability to effectively define disease status in ovarian cancer after initial therapy or to selectively screen high-risk populations remains a major challenge for in vivo monoclonal antibody (mAb)-targeted approaches. Antitumour murine mAbs (HMFG1, HMFG2, H317, and H17E2) and the reshaped human antibody Hu2PLAP (against placental alkaline phosphatase; PLAP), labelled with indium-111 and iodine-123, were evaluated for their ability to localise ovarian tumours in sequential studies of our group. Thirty patients with ovarian cancer, aged 40-78 years (median 60 years) were studied with HMFG1/G2: 11, and H317/H17E2: 19 murine mAbs. Six patients with ovarian cancer aged between 36 and 65 years (median 49 years) were studied with the reshaped human Hu2PLAP mAb (5 patients) or the murine H17E2 mAb (2 patients) labelled with 111In via a new macrocyclic chelating agent (DOTA). One of these was imaged twice, with H17E2- and Hu2PLAP-DOTA-111In, respectively. In 20 out of 22 patients with radiologically measurable ovarian cancer, the presence of tumour was confirmed by the murine mAb scan and correlated well with the findings of conventional radiology diagnostic methods. One of these patients with a negative H17E2 scan and a large abdominal mass at laparotomy was found to have a PLAP-negative tumour on immunohistochemistry. Additionally, the antibody scan revealed the presence of active disease, confirmed at laparotomy/laparoscopy, in 6 out of 8 patients considered to be in clinical complete remission. Best images were obtained at 24 and 48 h after the 123I and 111In mAbs, respectively. Successful imaging with the reshaped human antibody, Hu2PLAP, was seen in 2 patients with PLAP-positive tumours. Antibodies to DOTA developed in 2 patients. In conclusion, immunolocalisation of ovarian tumours is feasible with both murine and reshaped human mAbs. The sensitivity and specificity of the method appear very high in this pilot study, and in view of the absence of toxicity, the diagnostic contribution of this approach should be evaluated prospectively. Given the low number of patients without surgically detectable disease in the present study, future investigations should include more patients with no evidence of disease in order to provide more meaningful estimates of specificity. PMID- 9732223 TI - Use of serum tumor markers for the diagnosis and follow-up of breast cancer. PMID- 9732224 TI - The possible prognostic significance of p53 immunostaining status of the primary tumor in patients developing local recurrence after breast-conserving surgery. AB - Prognostic factors for distant metastases in patients with local recurrence after breast-conserving treatment (BCT) were studied. Fifty-six patients who developed local recurrence after BCT were recruited from 18 key hospitals/institutes in Japan. All 10 patients whose primary tumors were DCIS fared well without evidence of distant failure for a median follow-up period of 57 months (range 41-72) after the local recurrence. Inflammatory local recurrence was observed in 5 patients whose prognosis was grave: 3 with concomitant distant metastases and 1 developing them 7 months later. In the remaining 41 patients with noninflammatory local recurrence, various clinicopathological factors including age, disease-free interval, histology of the primary and recurrent tumors, axillary lymph node status, estrogen and progesterone receptor, immunohistochemical staining of erbB2 and p53 protein were evaluated as prognostic factors. Only the p53 immunostaining status of the primary tumor was found to be a significant prognostic indicator for distant metastases; distant disease-free survival at 5 years after the local recurrence was 92% for patients with p53-negative cancers and 51% for those with p53-positive cancers (p < 0.05). PMID- 9732225 TI - Prognostic significance of 67-kDa laminin receptor expression in advanced gastric cancer. AB - The ability of cancer cells to attach to laminin has been correlated with their metastatic potential and highly metastatic cancer cells seem to express on their surface significantly more laminin receptors than do their much less metastatic or benign counterparts. The expression of 67-kDa laminin receptors (LR) was investigated in a group of 75 patients who underwent gastrectomy for advanced gastric cancer, with special reference to the possible role in the tumor progression and in the overall survival. The tumor LR expression was immunohistochemically determined in paraffin-embedded sections using the MLuC5 monoclonal antibody which recognizes the 67-kDa LR and the avidin-biotin immunoperoxidase method. Of the 75 cases analyzed, 43 cases (57.3%) displayed a positive reaction. The cumulative 5-year survival rate was 72.6% (95% CI 52.5 85.3) for patients without expression of LR and 46.6% (29.8-61.8) for those with positive LR expression. A significant association between LR expression and depth of tumor invasion (0.022) was found. By univariate analysis the presence of laminin receptors seemed to be associated with a higher risk of death [RR 1.72 (95% CI 0.71-4.20], but this effect disappeared after controlling for depth of tumor invasion. In conclusion, these results suggest that tumor expression of laminin receptors could be correlated with tumor aggressiveness. However, the prognostic significance of laminin receptor expression is already provided by the depth of tumor invasion. PMID- 9732226 TI - Association between tumor angiogenesis and Borrmann type 4 carcinomas of the stomach. AB - A total of 202 gastric carcinomas, including 33 type 4 carcinomas, were studied. Intratumoral vessels were stained with an anti-CD34 monoclonal antibody before being quantitated by light microscopy (x200). The mean vessel counts for type 4 carcinomas were significantly higher than those for any other type of carcinoma, with the exception of type 1 carcinomas. Multivariate logistic regression showed that the vessel count was one of independent variables associated with type 4 carcinomas. In addition, survival analysis demonstrated that patients with type 4 carcinoma had significantly less favorable survival rates than those with other types of carcinomas. PMID- 9732227 TI - Putative role of dihydropyrimidine dehydrogenase in the toxic side effect of 5 fluorouracil in colorectal cancer patients. AB - Dihydropyrimidine dehydrogenase (DPD) is the first and rate limiting enzyme in the catabolism of 5-fluorouracil (5-FU). It has been reported from various laboratories that the plasma concentration of 5-FU was influenced by DPD activities in various normal human organs (e.g. liver or lymphocytes). Since the congenital deficiency in DPD caused severe, in some cases lethal, FU-related toxicity, it was decided to collect data about the DPD activity in colorectal cancer patients in order to investigate the possible correlation between the enzyme activity and appearance of the side effects of 5-FU. Assuming that DPD activity in lymphocytes represents the 5-FU catabolic capacity of the organism, DPD activity was determined in the lymphocytes of 48 patients with colorectal cancer after surgery during the therapeutic course with 5-FU and folinic acid. On the basis of the enzyme activity, patients were divided into three categories: low (DPD <5.03 pmol/min/10(6) lymphocytes); medium (DPD = 5.04-13.25 pmol/min/10(6) lymphocytes), and high (DPD > 13.26 pmol/min/10(6) lymphocytes) activity groups. By evaluating the toxic side effects during the 5-FU + folinic acid treatment, the following results were obtained. In the low DPD activity group, 9 of 11 patients had 5-FU-related side effects (mucositis, diarrhea, myelotoxicity, angina pectoris, hypertension). In 3 patients, no change of the therapy was needed, in 3 patients symptoms could be reversed by dose reduction of 5-FU while in 3 patients interruption of 5-FU therapy was needed. In the medium DPD activity group, mild toxicity (diarrhea, transitory hypertension) occurred in 5 of 29 and in the high activity group (diarrhea) in 1 of 8 patients, respectively. In these last two groups, no dose reduction of 5-FU was necessary. The present study furnished further evidence for the possible correlation between the 5-FU side effects and DPD function. Consequently, it is recommended to measure DPD activity prior to 5-FU based chemotherapy, which might be helpful in avoiding drug-related toxicity by adjusting the dose of 5-FU individually. PMID- 9732228 TI - Amplification of the c-met, c-erbB-2 and epidermal growth factor receptor gene in human gastric cancers: correlation to clinical features. AB - We examined amplification of the c-met, c-erbB-2, and epidermal growth factor receptor (EGFR) gene in the patients with primary gastric cancer, and compared the data with clinical features in order to clarify the relationship between oncogenic abnormality and clinical features. Oncogene amplifications were examined by slot blot hybridization using DNAs extracted from formalin-fixed and paraffin-embedded tissues of primary gastric cancers. Seven of the seventy cancers (10.0%) had c-met gene amplification, nine (12.9%) had c-erbB-2 gene amplification, and six (8.6%) had EGFR gene amplification, respectively. Eighteen cases (25.7%) exhibited one or multiple oncogene amplification, and two cases (2.9%) exhibited simultaneous amplification of the three genes. The cases with c met gene amplification tend to show invasive character and were related to peritoneal dissemination. The cases with c-erbB-2 gene amplification were related to lymph node metastasis. The cases with EGFR gene amplification had large tumors and were in highly advanced stage. The survival rate in patients with oncogene amplification was significantly lower than that in patients without amplification. Our data indicated that these genes were related to growth and metastasis of gastric cancer. Furthermore, this study about the three genes suggested that the type of activated gene might decide on the type of metastasis and clinical features. PMID- 9732229 TI - Characteristics of non-Hodgkin's lymphoma complicated by renal cell malignancies. AB - Previous epidemiological studies showed an increased risk for the development of renal cell carcinoma (RCC) after adjuvant therapy or simultaneous discovery of non-Hodgkin's lymphoma (NHL). However, clinicopathological features of NHL complicated by RCC are not well known. We summarized the clinicopathological features of 42 cases with malignant lymphoma complicated by renal tumors collected in a nationwide study in Japan. There were 27 males and 15 females, and the age at initial diagnosis of NHL ranged from 51 to 85 years (median 69 years). The clinical stages of NHL were stage I in 13 patients, stage II in 8, stage III in 8 and stage IV in 13 patients. RCC was simultaneously detected in 4 patients, within 1 year after the diagnosis of NHL in 20 and after more than 1 year in 13 patients. In the remaining 3 patients, NHL was diagnosed 1 year (2 cases) or 6 years (1 case) after nephrectomy. Histologically, all but 2 cases of NHL were diagnosed as diffuse lymphoma, with the large-cell type being the most common. The remaining 2 cases were follicular lymphoma. NHL in 35 cases (85%) were immunophenotyped as B cell type and 4 (10%) as T cell type. Renal tumors in all but 2 cases were classified as RCC: clear-cell type in 29, chromophobic type in 3, chromophilic type in 7 and Bellini duct carcinoma in 1 case. All RCC showed a cellular malignancy of grade II or III. When compared to sporadic autopsy cases of NHL in Japan, the frequency of extranodal lymphoma and B cell type was higher in the current cases (p = 0.06). In addition, leiomyosarcoma occasionally occurred among complicated renal tumors. PMID- 9732230 TI - Marked elevation of serum CA19-9 and stomach carcinoma. PMID- 9732231 TI - Italian guidelines for the management of infectious complications in pediatric oncology: empirical antimicrobial therapy of febrile neutropenia. AB - The Italian Association for Paediatric Haematology and Oncology prepared a guideline document aimed at unifying and rationalising as much as possible the management of febrile neutropenia in children with cancer, because of the potential impact of these procedures on hospital costs and on the development of antibiotic resistance. Before starting anti-infective therapy, at least 2 blood cultures, a throat swab, urine-culture, and cultures from any suspected infected site, should be performed. Routine chest X-rays at onset of febrile neutropenia are probably not necessary, in absence of respiratory signs. At the present time, the safer option probably remains the combination of a beta-lactam and an aminoglycoside, and treating febrile neutropenia outside of hospital should be considered an investigational approach. The choice of the most appropriated regimen for each institution should be based also on the local bacteriological statistics and patterns of bacterial resistance. Antibiotic toxicity and cost should be other important factors. Every subsequent addition or substitution of antibiotics should be based on objective signs of clinical deterioration. The only accepted empirical modification is empirical antifungal therapy, while the empirical addition of a glycopeptide antibiotic cannot be recommended. PMID- 9732232 TI - Regulation of the human glut4 gene expression in tumor RD18 cell line. AB - We tested if glut4, the gene for muscle-specific glucose transporter, underwent some variations of expression in neoplastic cells. Our model was a rhabdomyosarcoma cell line (RD18) which retains the ability to differentiate along the myogenic pathway. Any definable changes of expression of glut4 in normal and RD18 cells were revealed by Northern blot analysis. In order to identify the transcriptional regulatory regions of the glut4 gene we performed a deletion analysis of the 5' flanking region. The downregulation which we found in the expression of this gene in RD18 cells could be related with the activity of a negative regulatory element. PMID- 9732233 TI - Changes in matrix metalloproteinases during the evolution of interstitial renal fibrosis in a rat experimental model. AB - The aim of the present study was to analyze the matrix metalloproteinase (MMP) activity during the evolution of interstitial renal fibrosis in a rat experimental model of unilateral ureteral obstruction. The interstitial type I collagenase and the gelatinolytic activities were analyzed by radiolabeled substrate degradation. Interstitial collagenase activity was low at all times while gelatinolytic activity increased on day 6 of evolution, with a decrease in activity from this point. The use of organomercurials revealed the presence of latent enzyme in all cases. Normal kidney samples contained MMP-9 in both active and proenzyme forms as revealed by zymography. On day 3 MMP-9 dimers appeared, and increased activity was observed until day 6. A decrease in the gelatinolytic activity was detected from days 9-15 of evolution. This observation was confirmed by Western blot analysis that revealed the presence of proMMP-9 mainly from days 6-12. Tissue inhibitor of metalloproteinase-1 (TIMP-1) was also detected alone and in combination with proMMP-1 and MMP-1, particularly from days 6-15 of evolution. The presence of MMP-9 and MMP-1 was detected in the cytoplasm of cortical tubular cells by immunohistochemistry, with no difference between the experimental and the normal kidneys. There was also an increase in collagen concentration from day 3 after surgery that increased during the entire evolution of the experimental model. This work reveals that the decrease in the MMP-9 and MMP-1 enzymatic activity, due to their interaction with TIMP-1 and to the lack of activation of the latent forms, may participate in the excessive collagen deposit during the evolution of experimental interstitial renal fibrosis. PMID- 9732234 TI - The effect of interferon-gamma and tumor necrosis factor-alpha on the expression of ICAM-1 and HLA-DR molecules on cells of a human germ cell neoplasm and their susceptibility to lysis by lymphokine-activated killer cells. AB - The ICAM-1 molecule plays a role in the interaction of NK cells with a variety of tumor cells, including carcinoma, melanoma and glioblastoma cells. In the present study, we analyzed the effect of IFN-gamma and TNF-alpha on both the expression of HLA-DR and ICAM-1 molecules on HGCN (Germa-2), and on their susceptibility to lysis by LAK cells. Our results show that 1,000 U/ml IFN-gamma induced a substantial increase in the expression of both ICAM-1 molecules and HLA-DR on the cell surface, while the effect of TNF-alpha on the expression of these molecules was substantially less prominent. When Germa-2 cells, previously exposed to 1,000 U/ml IFN-gamma, were employed as target cells in a 4-hour 51Cr release assay, a statistically significant increase in the lysis by LAK cells was noted. These results show that in the presence of IFN-gamma, Germa-2 tumor cells undergo modulation which affects both the expression of ICAM-1 and HLA-DR molecules as well as their susceptibility to lysis by LAK cells. PMID- 9732235 TI - Tissue-specific high-level expression of human endogenous retrovirus-R in the human adrenal cortex. AB - In an attempt to clarify the biological nature of a human endogenous retrovirus (HERV), HERV-R, which is a single-copy type of HERVs and is conserved as a full length viral sequence, the expression of HERV-R mRNA in normal autopsied systemic organs was examined by Northern blot analysis. The expression showed different levels among individuals, with the adrenal glands expressing the highest level of HERV-R among all organs tested, except for the placenta. In various adrenal tumors, HERV-R was expressed at high levels in all cortical adenomas but less so in pheochromocytomas. In situ hybridization revealed the expression of HERV-R to be localized in all layers of the adrenal cortex, but not in the medulla. This high-level expression of HERV-R in the adrenal cortex may possibly relate to differentiation and/or steroid production by adrenocortical cells. PMID- 9732236 TI - Fluorescent in situ hybridization for identifying cytogenetic abnormalities in inadequate and suboptimal specimens. AB - Conventional cytogenetic analysis is often hampered by its dependency on the evaluation of dividing cells. When the mitotic index is low, or the cytogenetic preparation suboptimal, an accurate diagnosis often cannot be achieved using standard GTG banding. Molecular cytogenetics using fluorescent in situ hybridization (FISH) can be extremely useful in such a situation because it can be performed on nondividing interphase nuclei, and can usually be carried out on existing slides without requiring an additional specimen. The present paper presents two case histories to illustrate the utility of FISH in the clinical setting. PMID- 9732237 TI - Cell proliferation and apoptosis of the glomerular epithelial cells in rats with puromycin aminonucleoside nephrosis. AB - Injury and repair of the glomerular epithelial cells (GECs) play an important role in the pathogenesis of focal segmental glomerulosclerosis (FSGS). To obtain a better understanding of proliferation and apoptosis of GECs, we examined immunohistochemical and in situ hybridization findings in puromycin aminonucleoside nephrosis (PAN) of rats. The minimal-change nephrotic syndrome model (PAN-MCNS) was induced by administering 5 subcutaneous injections of puromycin aminonucleoside (PA; each 1.5 mg/100 g B/W to one group of rats), whereas the FSGS model (PAN-FSGS) was induced by administering an additional 5 injections of PA to another group of rats. The cell kinetics of the GECs were assessed with labeling 5-bromo 2'-deoxyuridine (BrdU) and proliferating cell nuclear antigen (PCNA). To investigate regulation of apoptosis in rats with PAN, we evaluated the expression of p53, Fas antigen, Fas ligand and Bc1-2. Rats with PAN-MCNS exhibited a significantly greater number of BrdU- and PCNA-labeled GECs as compared with control rats. In rats with PAN-FSGS, the number of PCNA-labeled GECs was greater than in rats with PAN-MCNS, but the number of BrdU-labeled GECs was lower. Apoptotic cells were occasionally observed in the sclerotic lesions, with the number being significantly higher in rats with PAN-FSGS than in rats with PAN-MCNS and control. Apoptotic cells were observed in the GECs of PAN-FSGS rats. However, they were negative for p53, Fas antigen, and Fas ligand. Immunohistochemical and in situ hybridization studies revealed a greater intraglomerular overexpression of Bc1-2 protein and bc1-2 mRNA in the PAN-FSGS rats as compared with control rats. These results suggest that insufficient proliferation and apoptosis in GECs may be involved in the progression of FSGS. PMID- 9732238 TI - Prolonged hematopoietic chimerism in normal mice transplanted in utero with human hematopoietic stem cells. AB - We have previously reported prolonged hematopoietic chimerism in normal mice transplanted in utero with human fetal hematopoietic stem cells (HSC) by flow cytometry. We now further confirm the human origin of these cells by demonstrating human DNA in the marrow of one such chimeric mouse. We also examined 42 mice born after in utero transplantation with HSC enriched from human adult marrow cells. All live-born mice were treated with recombinant human growth factors. Twelve had human cells in the peripheral blood (range: 01.-2.93%). Thymic samples were positive in 3 cases. The bone marrow of 2 mice contained cells expressing human CD34 antigen. Light scatter characteristics support the presence of multilineage hematochimerism. Human IgM was present in 2 of 4 chimeric sera tested. Thus, normal mice transplanted in utero with human HSC may permit long-term engraftment and differentiation of the human HSC. PMID- 9732239 TI - High-risk human papillomavirus infection and E6 protein expression in lesions of the uterine cervix. AB - Pathologic and epidemiologic investigations carried out over the past several years have provided evidence that carcinogenesis in the uterine cervix is a multi step process involving discreet preinvasive stages. Molecular epidemiologic data also indicate that human papillomavirus (HPV) infection is a critical factor in the tumor progression process. In vitro studies have shown that for the initiation and maintenance of the malignant phenotype, the expression of the HPV transforming protein E6 is required. The E6 protein produced by the high-risk HPV types 16 and 18 can bind to and inactivate the tumor suppressor protein p53 leading to deregulated proliferation and defective apoptosis, thus facilitating tumor progression. Therefore, determination of the HPV genotype alone may not be sufficient in assessing tumor progression in the uterine cervix. In the present study, a total of 623 cervical tissue samples at various phases of tumor progression were assessed for HPV infection by nonisotopic in situ hybridization (NISH) and for HPV 16/18 E6 protein expression by immunocytochemistry. There was significant correlation between the extent of histological abnormality and HPV infection. Significant correlation (r = 0.707, p = 0.000) was observed between the presence of HPV 16 and high-grade squamous intraepithelial lesions (SILs) and invasive cancer. The odds ratio of a cervical tissue infected with HPV 16 falling into these two categories was 44.57 (95% CI: 27.10, 73.30). The E6 protein also was mostly detected in high-grade SILs and cervical cancer tissue expressing either HPV 16 or 18. It was less frequent in low-grade SILs infected with HPV 16/18 and was absent in benign cervical tissue infected with HPV 16. The odds ratio of an HPV-16/18-infected cervical tissue positive for E6 being a high-grade SIL or invasive cancer was 16.20 (95% CI: 6.06, 43.33). These results thus show the clinical utility of HPV characterization along with the analysis of the transforming protein E6 in the assessment of tumor progression in the uterine cervix. PMID- 9732240 TI - Regulation of vascular endothelial growth factor receptor KDR in vitro by a soluble factor in confluent endothelial cells. AB - Hypoxia regulates the expression of both vascular endothelial growth factor (VEGF) and its receptor (KDR). We have shown that cell density regulates VEGF expression in colon cancer and hypothesized that a similar mechanism regulates KDR in endothelial cells. Human umbilical vein endothelial cells were grown as sparse and confluent monolayers. Northern blot analysis revealed that KDR and VEGF mRNA expression in confluent cells was more than two-fold greater than in sparse cells. In contrast, flt-1 expression increased only slightly in cells grown to confluence. Cells were then plated at various concentrations and subjected to semi-quantitative PCR; KDR mRNA expression increased as cell density increased. Serum-free conditioned medium from cells grown to confluency for 48 h was added to sparsely plated cells, and KDR expression in the sparse cells increased twofold. We conclude that cell density regulates KDR endothelial cell expression via an unidentified soluble factor. PMID- 9732241 TI - Severe pediatric head injury: myth, magic, and actual fact. AB - Reports of low mortality rate for severely head-injured children have led to the myth that children have a better outcome than adults after severe head injury. This was examined in the present study by comparing the outcome of 4,041 severely head-injured children with 14,789 adults from five Level I trauma centers in the Pennsylvania Trauma Systems Foundation from 1986 to 1996. Severe head injury was defined as GCS <8, and outcome was defined by mortality. The overall mortality rate for children (<18 years) was 36.5% and for adults 47. 6%. However, for patients involved in motor vehicle accidents, there was no clinically significant difference between children age 3-11 years (35%), age 12-18 years (31.4%) and adults (32.5%). The unprecedented low mortality rate for severe head injury in children was probably due to the selection bias of the patient population. We conclude that children involved in motor vehicle accidents are just as likely to die from severe head injury as adults. PMID- 9732242 TI - Death in shunted hydrocephalic children in the 1990s. AB - Using a combined search of the Children's Hospital (Birmingham, Ala., USA) medical records and the Jefferson County Health Department death records, we reviewed all shunt-related deaths that occurred between January 1990 and July 1996. Of these, we excluded patients who died of nonhydrocephalus-related reasons, such as bronchopulmonary dysplasia, as well as patients who had other serious neurological illnesses such as brain tumor and hydranencephaly. Twenty eight patients died of shunt-related causes in the 6.5-year period. A survival analysis showed that 96% survived 32 months after first shunting. Of 28 patients, 23 were beyond help prior to medical evaluation. However, at least 10 of these patients had symptoms suggestive of shunt failure at least 24 h and as long as 2 weeks prior to their demise. We conclude that hydrocephalic children still die of shunt failure despite the modern technology of the 1990s. Some of these causes may be avoidable through early detection of symptoms. Guidelines to patients, families, and primary caregivers should be emphasized. PMID- 9732243 TI - Myelomeningocele repair in utero: a report of three cases. AB - BACKGROUND: Evidence accumulating over the last 10 years suggests that the exposed spinal cord tissue in a myelomeningocele sustains a secondary injury as the result of prolonged exposure to the intrauterine environment. These data suggest that early closure of the myelomeningocele sac might prevent such injury and in turn improve the neurologic outcome in the affected infant. METHODS: Three patients with fetuses carrying the ultrasonic diagnosis of myelomeningocele elected to enter a study of the feasibility of repairing myelomeningocele in utero. At approximately 28 weeks of gestation each patient underwent laparotomy and hysterotomy, thus exposing the myelomeningocele defect. The defect was closed in a routine surgical fashion, and the hysterotomy was then closed. RESULTS: The 3 patients recovered from surgery without incident. Early premature contractions subsided, and they were discharged by the 5th postoperative day. At between 33 and 36 weeks of gestation, each infant was delivered via cesarean section. The observed neurologic deficits were within the range expected from the anatomic level of the defects. Two of the infants have not required ventriculoperitoneal shunting. CONCLUSIONS: This limited series of patients suggests that myelomeningocele can be repaired in utero with minimal morbidity to either the mother or her fetus. A larger study will be needed to substantiate this low morbidity and to determine the extent of any neurologic benefit of early surgery. PMID- 9732244 TI - Abnormalities of water metabolism after surgery for optic/chiasmatic astrocytomas in children. AB - A major concern during the early postoperative period after surgical resection of optic chiasmatic gliomas is the derangement of sodium and water metabolism which may add to the morbidity of the procedure. The purpose of this study was to characterize the abnormalities of water and sodium metabolism in children with optic chiasmatic gliomas treated surgically at British Columbia's Children's Hospital and to identify therapeutic modalities which might prevent or ameliorate the development of these complications. A retrospective chart review of children with optic/chiasmatic gliomas undergoing operations on the tumor was performed and the pre- and postoperative radiographs reviewed by an independent neuroradiologist. There were 11 patients who underwent 13 operations for either resection (n = 9) or biopsy (n = 4) of their optic chiasm tumor. The extent of resection in patients undergoing more than simple biopsy ranged from 83 to 99%, and all patients with resection had exophytic tumor extending into the hypothalamus. Postoperative syndrome of inappropriate antidiuretic hormone secretion (SIADH) and/or diabetes insipidus occurred after 8 of the 9 tumor resections and was associated with significant morbidity. No disturbance of water metabolism occurred after biopsy only. In patients with SIADH, the urinary sodium level rose markedly 6-12 h prior to the development of hyponatremia, and it was concluded that this was a valuable predictor of impending hyponatremia. Replacement of urine output was noted to be the most reliable way to avoid rapid fluctuations in serum sodium and to avoid the morbidity of diabetes insipidus or SIADH and is recommended in the postsurgical patients who cannot regulate fluid intake. PMID- 9732245 TI - The internet as a pediatric neurosurgery information resource. AB - The Internet is evolving as a powerful source of medical information for patients, family members, and health care providers; however, critical analyses of this information are generally lacking. This investigation was carried out to ascertain the sources, nature, and accuracy of information acquired via searches on the Internet for ten common pediatric neurosurgical topics. Thirteen source types and nine categories of information were characterized and examined. With few notable exceptions, the educational information was highly accurate. It is concluded that the Internet represents an important new pediatric neurosurgery educational resource for the layman and health care providers. Guidelines are offered to assist in the use of this new medium. PMID- 9732246 TI - Untethered filum terminale presenting as cauda equina lesion. AB - We report the case of a young girl with recurrent bladder dysfunction. Magnetic resonance imaging performed for evaluation of initial urologic symptoms revealed a low-lying conus medullaris. She underwent an L5 laminectomy and cord untethering by sectioning of the filum terminale. After initial improvement of bladder function, her symptoms returned 4 years later. Repeat magnetic resonance imaging demonstrated a new intradural lesion at L2. At surgery she was found to have an untethered, thickened, coiled filum terminale at L2. PMID- 9732247 TI - Primary hypothyroidism mimicking a pituitary macroadenoma. AB - Pituitary macroadenomas occur infrequently in children and can present with visual dysfunction or endocrinopathy. Occasionally, primary end-organ failure can cause reactive enlargement of the pituitary gland that may be clinically and radiographically indistinguishable from pituitary macroadenoma. The authors describe an example of reactive pituitary hyperplasia from primary hypothyroidism that mimicked a pituitary macroadenoma in a child. Recognition of this entity, which responds to medical therapy, is important because it can spare patients unnecessary surgery. PMID- 9732248 TI - Unusual association of intractable temporal lobe seizures and intracranial aneurysms in an adolescent: is it a coincidence? AB - Intracranial aneurysms in the pediatric age group are rare occurrences. They usually present with subarachnoid hemorrhage or mass effect. Their association with epilepsy has rarely been reported; such concurrence may not be a coincidence. We present a 16-year-old girl with a 5-year history of medically intractable complex partial seizures. Preoperative electrophysiological and neuroimaging studies demonstrated an epileptogenic focus and atrophy in the right mesial temporal lobe, and ipsilateral incidental aneurysm at the carotid artery bifurcation. The patient underwent a complete right anterior temporal lobectomy, followed by clipping of the aneurysm. We concluded that the epilepsy management in association with cerebral aneurysms is controversial, but when surgery is indicated, clipping of the aneurysm and resection of the epileptogenic focus may provide the optimal outcome. The relevant literature is reviewed and the possible mechanisms of production of epilepsy by intracranial aneurysms are discussed. PMID- 9732249 TI - Myelitis: a rare presentation of mumps. AB - We report a rare case of mumps myelitis in which parotid swelling appeared 3 days after the symptoms of myelitis. A 10-year-old boy presented with acute paraplegia of grade I-II on MRC (Medical Research Council) scale and retention of urine with normal sensations. Central motor conduction to tibialis anterior (CMCT-TA) on the right side was 32 ms and 24 ms on the left side. Spinal MRI revealed hyperintense signal changes extending from C3 to T12. After 5 days of methylprednisolone therapy, there was marked improvement in weakness, micturition normalized and CMCT-TA also returned to normal. MRI repeated after 1 month was also normal. This response seems to be due to antiedema and to the antiinflammatory effect of methylprednisolone, because of a temporal relationship between MPS and clinical improvement. However, the possibility of natural recovery cannot be ruled out. PMID- 9732250 TI - Back pain in an 8-year-old boy. PMID- 9732251 TI - Urbach-Wiethe disease (Lipoid proteinosis). PMID- 9732252 TI - Prognostic factors in infants and very young children with intracranial ependymomas. AB - The Pediatric Oncology Group (1986-1990) conducted a study in which 48 children <3 years of age with intracranial ependymomas were treated with prolonged postoperative chemotherapy (CT) and delayed RT. Thirty-one children, 0-23 months of age at diagnosis (Gp A) received 2 years of CT followed by RT; while 17 children, 24-36 months of age at diagnosis (Gp B) received CT for 1 year followed by radiation. One-year survivals were 87% (Gp A) and 94% (Gp B) and 2-year survivals were 67% (Gp A) and 82% (Gp B). In subsequent years a significant divergence in survivals according to age has been noted (p = 0.04). Five-year survivals were 25.7% (Gp A) vs. 63.3% (Gp B). The curves began to diverge 1 year following diagnosis. Other than age, the only significant prognostic factor was degree of surgical resection: 5-year survivals were 66% (total resection) vs. 25% (subtotal resection). Neither the presence of metastases, degree of anaplasia nor the degree of surgical resection varied significantly according to age at diagnosis. The most likely reason for the difference in survivals between the two age groups relates to the timing of radiation following CT, i.e., 1-year delay in children 24-36 months of age compared to a 2-year delay in children 0-23 months of age. An alternative but less likely hypothesis is that ependymomas in the younger children have a more aggressive biology. In contrast, survivals in the 24 to 36-month group are much better than previous reports in the literature suggesting that prolonged postoperative CT may allow both a delay in CRT as well as provide improved survivals. Based on these results, future treatment trials should emphasize maximal surgical resection and a delay in radiation of no more than 1 year. PMID- 9732253 TI - A novel embryogenetic mechanism for Currarino's triad: inadequate dorsoventral separation of the caudal eminence from hindgut endoderm. AB - Currarino's triad is a congenital malformation involving the combination of anorectal stenosis, a presacral mass (most often a teratoma or ventral menigocele) and an anterior sacral bony defect (scimitar sacrum). Current theories regarding its embryogenesis are difficult to reconcile with our current understanding of caudal neuraxial and hindgut development. Caudal neuraxial structures develop from the caudal eminence (or tail bud), which normally separates from the hindgut endoderm concurrent with ingrowth of the posterior notochord during late gastrulation. We describe the first reported association of Currarino's triad with a caudal split cord malformation. It has previously been proposed that split cord malformations and related 'complex dysraphic malformations' involving abnormalities of one or more of the three primary germ layers arise through disordered midline axial integration during gastrulation. The presence of a split cord malformation in a patient with Currarino's triad suggests that the two disorders share a common embryogenetic pathway. We propose that the malformations of Currarino's triad arise through a failure of dorsoventral separation of the caudal eminence from the hindgut endoderm during late gastrulation. PMID- 9732254 TI - Dural tenting sutures in pediatric neurosurgery. AB - Dural tenting sutures were used on a selective basis in 329 consecutive cranial operations in children; 16 (4.8%) required dural tenting sutures for the control of epidural bleeding and 313 (95.2%) required no dural sutures. Dural tenting sutures were never placed for prophylaxis or to satisfy a routine. Reoperation for postoperative epidural hematoma was required in 1 child (0.3%) and that child was subsequently discovered to have hemophilia. The author concludes that there is no compelling evidence for the prophylactic use of dural tenting sutures in children. PMID- 9732255 TI - Endoscopic third ventriculostomy: an outcome analysis. AB - Endoscopic third ventriculostomy (ETV) has gained widespread acceptance as an effective way to manage hydrocephalus in selected patients. To determine which patient groups have the highest chance of successful ETV, a retrospective case review was performed. From June 1992 to December 1996, 97 patients underwent a total of 98 ETVs at our institution. There were 59 males and 38 females with a mean age of 8.1 years (range 1 day to 29.5 years). Twenty-six of 98 procedures (26%) were abandoned due to either unfavorable anatomy, inability to perform a cisternostomy, or hemorrhage. Follow-up data was available in 92 patients for a mean of 24.2 months. The rate of successful ETV in 71 patients, with either complete shunt avoidance or removal, varied widely by diagnosis and patient age. The highest success rates were achieved in patients with aqueductal stenosis, tectal plate tumor, myelomeningocele and posterior fossa tumor. Complications included one transient herniation syndrome, one basilar artery perforation, 2 cases of ventriculitis, one transient decrease in level of consciousness, and one transient hemiparesis. We feel these results support the continued use of ETV in only carefully selected patients with hydrocephalus. PMID- 9732256 TI - Treatment and long-term outcome of pineal nongerminomatous germ cell tumors. AB - In 1994 we reported on a small series of 3 children with malignant, marker positive pineal nongerminomatous germ cell tumors treated with a 'sandwich' protocol. Here, we report on the long-term survival of these children. Preoperative chemotherapy consisted of two courses of bleomycin, etoposide, and cisplatin. En bloc resection of the tumors via the supracerebellar, infratentorial route was performed immediately after decline of tumor marker levels. Postoperatively, two courses of vinblastine, ifosfamide, and cisplatin were applied, followed by craniospinal irradiation. The patients showed no major neurological deficits and no evidence (neuroradiologically or with regard to tumor marker levels) of recurrence of disease after 66, 71, and 78 months, respectively. We propose this regimen for children with tumors of the pineal region in whom the tumor markers are positive. It should be started without histological classification of the tumor to avoid possible spillage of malignant tumor cells to the cerebrospinal fluid. PMID- 9732257 TI - Effects of proteins, blood cells and glucose on the viscosity of cerebrospinal fluid. AB - It has long been assumed that cerebrospinal fluid (CSF) is a newtonian fluid with viscosity similar to water, yet high protein content, has been postulated to increase the viscosity of CSF in vivo. Such an increase in viscosity may have serious implications for the effectiveness of surgical shunts implanted to re establish the CSF flow in cases of abnormal CSF circulation. In this study, glucose content, total protein content and blood cell count in the CSF of 23 patients undergoing brain surgery were measured. Viscosity measurements were performed on duplicate CSF samples over a range of shear strain rates of 25-1,460 s-1. The results indicated that high protein or high cell concentration in CSF does not significantly affect the viscosity of the cerebral fluid at those shear rates. CSF is clearly newtonian, and its viscosity at 37 degreesC is in the range of 0.7-1 mPa.s. PMID- 9732258 TI - Treatment of villous hypertrophy of the choroid plexus by endoscopic contact coagulation. AB - Villous hypertrophy of the choroid plexus (VHCP) is a condition characterized by overproduction of cerebrospinal fluid by bilaterally symmetric and enlarged, yet histologically normal, choroid plexi. Medical and surgical therapy have been met with variable success. Traditional shunting procedures often result in failure to correct the underlying problem as well as failure to absorb the large volume of fluid produced. Craniotomy for open surgical resection of the choroid is associated with considerable morbidity. The authors describe a technique of endoscopic contact coagulation as an effective and safe treatment of VHCP. A 14 month-old hydrocephalic girl with VHCP who failed ventriculoperitoneal shunting as the sole treatment of her hydrocephalus presented with increasing ascites. She was successfully treated with ventriculoperitoneal shunting and endoscopic Bugby wire-directed monopolar contact coagulation of the hyperplastic choroid plexus. Postoperatively her head circumference and cognitive development is normal for her age, and her ascites has resolved. Endoscopic contact coagulation of the hyperplastic choroid plexus is a minimally invasive surgical method which treats the cause of the CSF production while avoiding the operative complications of open choroid plexus resection. PMID- 9732259 TI - Intramedullary lipoma of dorsocervicothoracic spinal cord with intracranial extension and hydrocephalus. AB - An intradural, intramedullary lipoma originating within the cervicothoracic cord with significant extension into the posterior fossa is reported. The lipoma was found incidentally by CT scan at the infant's age of 2 days. Follow-up studies by CT and MRI were performed on several occasions. Because of hydrocephalus, VP shunting was performed. After shunting and shunt revisions, surgery of lipoma was performed at the age of 2 years. Posterior fossa craniectomy and laminectomy from C1 to T4 revealed a lipoma, which was maximally reduced. Six months after surgery, the child was well and showed good recovery. PMID- 9732260 TI - Focal cortical dysplasia with glioproliferative changes causing seizures: report of 3 cases. AB - In contrast to neoplasia, lesions of focal cerebral dysplasia are thought to be completed developmental processes of abnormal neuronal migration. We present three children with seizures resulting from brain lesions which pathologically demonstrate regions of both clearcut focal cortical dysplasia and also hypercellularity and monomorphism typical of proliferative lesions such as low grade glial tumor. These cases suggest the existence of a distinct subgroup of patients with prominent glioproliferative changes in association with focal cortical dysplasia, challenging the conventional dichotomy between dysplastic and proliferative categories of brain lesions. Recognition of patients with dual pathology may be of practical as well as theoretical importance. PMID- 9732261 TI - Five-year-old female with a cyst of the fourth ventricle. PMID- 9732262 TI - The role of chemotherapy in newly diagnosed ependymoma of childhood. AB - The use of chemotherapy in treating children with newly diagnosed ependymoma is currently being investigated, both clinically and experimentally. Assessment of the true efficacy of this modality is hampered by a lack of prospective randomized trials comparing conventional treatment schemes of aggressive surgical excision followed by radiation therapy with or without addition of chemotherapy. Although with current regimens, the role of chemotherapy in newly diagnosed disease appears limited, measurable disease responses have been recognized. Preliminary studies support the potential use of chemotherapy in infants in an effort to delay radiation therapy or even avoid radiation therapy, although with conventional chemotherapy, a delay exceeding 12 months is inadvisable. Additionally, preliminary data suggest that in children with incompletely resected tumors, chemotherapy may be of benefit as an adjunct to second-look surgery. Experimental work investigating chemotherapeutic sensitivity, molecular genetics, or mechanisms to overcome drug resistance may offer some benefit in utilizing chemotherapy for ependymoma of childhood. PMID- 9732263 TI - LMO T-cell translocation oncogenes typify genes activated by chromosomal translocations that alter transcription and developmental processes. PMID- 9732264 TI - JNK targets p53 ubiquitination and degradation in nonstressed cells. AB - In this study we elucidated the role of nonactive JNK in regulating p53 stability. The amount of p53-JNK complex was inversely correlated with p53 level. A peptide corresponding to the JNK binding site on p53 efficiently blocked ubiquitination of p53. Similarly, p53 lacking the JNK binding site exhibits a longer half-life than p53(wt). Outcompeting JNK association with p53 increased the level of p53, whereas overexpression of a phosphorylation mutant form of JNK inhibited p53 accumulation. JNK-p53 and Mdm2-p53 complexes were preferentially found in G0/G1 and S/G2M phases of the cell cycle, respectively. Altogether, these data indicate that JNK is an Mdm2-independent regulator of p53 stability in nonstressed cells. PMID- 9732265 TI - SNAP19 mediates the assembly of a functional core promoter complex (SNAPc) shared by RNA polymerases II and III. AB - The basal transcription factor SNAPc binds to the PSE, a core element in the RNA polymerase II and III human snRNA promoters. SNAPc contains at least four subunits, but it has not been possible to assemble a fully defined recombinant SNAPc. Here we reconstitute SNAPc from five recombinant subunits, SNAP43, SNAP45, SNAP50, SNAP190, and a newly identified subunit, SNAP19. This recombinant complex binds specifically to the PSE and directs both RNA polymerase II and III snRNA gene transcription. Thus, the same core SNAPc nucleates the assembly of two classes of initiation complexes. PMID- 9732266 TI - Phosphorylation regulates association of the transcription factor Pho4 with its import receptor Pse1/Kap121. AB - The transcription factor Pho4 is phosphorylated and localized predominantly to the cytoplasm when budding yeast are grown in phosphate-rich medium and is unphosphorylated and localized to the nucleus upon phosphate starvation. We have investigated the requirements for nuclear import of Pho4 and find that Pho4 enters the nucleus via a nonclassical import pathway that utilizes the importin beta family member Pse1/Kap121. Pse1 binds directly to Pho4 and is required for its import in vivo. We have defined the nuclear localization signal on Pho4 and demonstrate that it is required for Pse1 binding in vitro and is sufficient for PSE1-dependent import in vivo. Phosphorylation of Pho4 inhibits its interaction with Pse1, providing a mechanism by which phosphorylation may regulate import of Pho4 in vivo. PMID- 9732267 TI - Membrane recruitment of the kinase cascade scaffold protein Ste5 by the Gbetagamma complex underlies activation of the yeast pheromone response pathway. AB - In the Saccharomyces cerevisiae pheromone response pathway, the Gbetagamma complex activates downstream responses by an unknown mechanism involving a MAP kinase cascade, the PAK-like kinase Ste20, and a Rho family GTPase, Cdc42. Here we show that Gbetagamma must remain membrane-associated after release from Galpha to activate the downstream pathway. We also show that pheromone stimulates translocation of the kinase cascade scaffold protein Ste5 to the cell surface. This recruitment requires Gbetagamma function and the Gbetagamma-binding domain of Ste5, but not the kinases downstream of Gbetagamma, suggesting that it is mediated by Gbetagamma itself. Furthermore, this event has functional significance, as artificial targeting of Ste5 to the plasma membrane, but not intracellular membranes, activates the pathway in the absence of pheromone or Gbetagamma. Remarkably, although independent of Gbetagamma, activation by membrane-targeted Ste5 requires Ste20, Cdc42, and Cdc24, indicating that their participation in this pathway does not require them to be activated by Gbetagamma. Thus, membrane recruitment of Ste5 defines a molecular activity for Gbetagamma. Moreover, our results suggest that this event promotes kinase cascade activation by delivering the Ste5-associated kinases to the cell surface kinase Ste20, whose function may depend on Cdc42 and Cdc24. PMID- 9732268 TI - CLB5 and CLB6 are required for premeiotic DNA replication and activation of the meiotic S/M checkpoint. AB - Initiation of DNA replication during the mitotic cell cycle requires the activation of a cyclin-dependent protein kinase (CDK). The B-type cyclins Clb5 and Clb6 are the primary activators of the S phase function of the budding yeast CDK Cdc28. However, in mitotically growing cells this role can be fulfilled by the other B-type cyclins Clb1-Clb4. We report here that cells undergoing meiotic development also require Clb dependent CDK activity for DNA replication. Diploid clb5/clb5 clb6/clb6 mutants are unable to perform premeiotic DNA replication. Despite this defect, the mutant cells progress into the meiotic program and undergo lethal segregation of unreplicated DNA suggesting that they fail to activate a checkpoint that restrains meiotic M phase until DNA replication is complete. We have found that a DNA replication checkpoint dependent on the ATM homolog MEC1 operates in wild-type cells during meiosis and can be invoked in response to inhibition of DNA synthesis. Although cells that lack clb5 and clb6 are unable to activate the meiotic DNA replication checkpoint, they do possess an intact DNA damage checkpoint which can restrain chromosome segregation in the face of DNA damage. We conclude that CLB5 and CLB6 are essential for premeiotic DNA replication and, consequently, for activation of a meiotic DNA replication checkpoint. PMID- 9732269 TI - okra and spindle-B encode components of the RAD52 DNA repair pathway and affect meiosis and patterning in Drosophila oogenesis. AB - okra (okr), spindle-B (spnB), and spindle-D (spnD) are three members of a group of female sterile loci that produce defects in oocyte and egg morphology, including variable dorsal-ventral defects in the eggshell and embryo, anterior posterior defects in the follicle cell epithelium and in the oocyte, and abnormalities in oocyte nuclear morphology. Many of these phenotypes reflect defects in grk-Egfr signaling processes, and can be accounted for by a failure to accumulate wild-type levels of Gurken and Fs(1)K10. We have cloned okr and spnB, and show that okr encodes the Drosophila homolog of the yeast DNA-repair protein Rad54, and spnB encodes a Rad51-like protein related to the meiosis-specific DMC1 gene. In functional tests of their role in DNA repair, we find that okr behaves like its yeast homolog in that it is required in both mitotic and meiotic cells. In contrast, spnB and spnD appear to be required only in meiosis. The fact that genes involved in meiotic DNA metabolism have specific effects on oocyte patterning implies that the progression of the meiotic cell cycle is coordinated with the regulation of certain developmental events during oogenesis. PMID- 9732271 TI - Pax9-deficient mice lack pharyngeal pouch derivatives and teeth and exhibit craniofacial and limb abnormalities. AB - Pax genes have been shown to play important roles in mammalian development and organogenesis. Pax9, a member of this transcription factor family, is expressed in somites, pharyngeal pouches, mesenchyme involved in craniofacial, tooth, and limb development, as well as other sites during mouse embryogenesis. To analyze its function in vivo, we generated Pax9 deficient mice and show that Pax9 is essential for the development of a variety of organs and skeletal elements. Homozygous Pax9-mutant mice die shortly after birth, most likely as a consequence of a cleft secondary palate. They lack a thymus, parathyroid glands, and ultimobranchial bodies, organs which are derived from the pharyngeal pouches. In all limbs, a supernumerary preaxial digit is formed, but the flexor of the hindlimb toes is missing. Furthermore, craniofacial and visceral skeletogenesis is disturbed, and all teeth are absent. In Pax9-deficient embryos tooth development is arrested at the bud stage. At this stage, Pax9 is required for the mesenchymal expression of Bmp4, Msx1, and Lef1, suggesting a role for Pax9 in the establishment of the inductive capacity of the tooth mesenchyme. In summary, our analysis shows that Pax9 is a key regulator during the development of a wide range of organ primordia. PMID- 9732270 TI - A novel homeobox gene mediates the Dpp signal to establish functional specificity within target cells. AB - Morphogen gradients of secreted molecules play critical roles in the establishment of the spatial pattern of gene expression. During midgut development in Drosophila, secreted molecules of Decapentaplegic (Dpp) and Wingless (Wg) establish unique transcriptional regulation within target cells to specify the resultant cell types. Here we report the identification of a novel homeobox gene, defective proventriculus (dve), which is required for the midgut specification under the control of Dpp and Wg. In dve mutants, two distinct parts of the midgut, the proventriculus and middle midgut, are abnormally organized. The Wg signal regulates dve expression during proventriculus development. On the other hand, dve is a downstream target of Dpp in the middle midgut and defines the functional specificity of copper cells along with another Dpp target gene, labial. Thus, the dve gene acts under the two distinct extracellular signals at distant parts of the midgut primordia. PMID- 9732272 TI - Transcription occurs in pulses in muscle fibers. AB - We report a novel mechanism of gene regulation in skeletal muscle fibers. Within an individual myofiber nucleus, not all muscle loci are transcriptionally active at a given time and loci are regulated independently. This phenomenon is particularly remarkable because the nuclei within a myofiber share a common cytoplasm. Both endogenous muscle-specific and housekeeping genes and transgenes are regulated in this manner. Therefore, despite the uniform protein composition of the contractile apparatus along the length of the fiber, the loci that encode this structure are not transcribed continuously. The total number of active loci for a particular gene is dynamic, changing during fetal development, regeneration, and in the adult, and potentially reflects the growth status of the fiber. The data reveal that transcription in particular stages of muscle fiber maturation occurs in pulses and is defined by a stochastic mechanism. PMID- 9732273 TI - Transcriptional termination in the Balbiani ring 1 gene is closely coupled to 3' end formation and excision of the 3'-terminal intron. AB - We have analyzed transcription termination, 3'-end formation, and excision of the 3'-terminal intron in vivo in the Balbiani ring 1 (BR1) gene and its pre-mRNA. We show that full-length RNA transcripts are evenly spaced on the gene from a position 300 bp upstream to a region 500-700 bp downstream of the polyadenylation sequence. Very few full-length transcripts and no short, cleaved, nascent transcripts could be observed downstream of this region. Pre-mRNA with 10-20 adenylate residues accumulates at the active gene and then rapidly leaves from the gene locus. Only polyadenylated pre-mRNAs could be detected in the nucleoplasm. Our results are consistent with the hypothesis that transcription termination occurs in a narrow region for the majority of transcripts, simultaneous with 3'-end formation. Excision of the 3'-terminal intron occurs before 3'-end formation in about 5% of the nascent transcripts. When transcription terminates, 3' cleavage takes place and 10-20 adenylate residues are added, the 3'-terminal intron is excised from additionally about 75% of the pre-mRNA at the gene locus. Our data support a close temporal and spatial coupling of transcription termination and the cleavage and initial polyadenylation of 3'-end formation. Excision of the 3'-terminal intron is highly stimulated as the cleavage/polyadenylation complex assembles and 3'-end formation is initiated. PMID- 9732274 TI - Ribonuclease E organizes the protein interactions in the Escherichia coli RNA degradosome. AB - The Escherichia coli RNA degradosome is the prototype of a recently discovered family of multiprotein machines involved in the processing and degradation of RNA. The interactions between the various protein components of the RNA degradosome were investigated by Far Western blotting, the yeast two-hybrid assay, and coimmunopurification experiments. Our results demonstrate that the carboxy-terminal half (CTH) of ribonuclease E (RNase E) contains the binding sites for the three other major degradosomal components, the DEAD-box RNA helicase RhlB, enolase, and polynucleotide phosphorylase (PNPase). The CTH of RNase E acts as the scaffold of the complex upon which the other degradosomal components are assembled. Regions for oligomerization were detected in the amino terminal and central regions of RNase E. Furthermore, polypeptides derived from the highly charged region of RNase E, containing the RhlB binding site, stimulate RhlB activity at least 15-fold, saturating at one polypeptide per RhlB molecule. A model for the regulation of the RhlB RNA helicase activity is presented. The description of RNase E now emerging is that of a remarkably complex multidomain protein containing an amino-terminal catalytic domain, a central RNA-binding domain, and carboxy-terminal binding sites for the other major components of the RNA degradosome. PMID- 9732275 TI - Sequence and DNA structural determinants of N4 virion RNA polymerase-promoter recognition. AB - Coliphage N4-coded, virion-encapsidated RNA polymerase (vRNAP) is able to bind to and transcribe promoter-containing double-stranded DNAs when the template is supercoiled and Escherichia coli single-stranded DNA-binding protein (Eco SSB) is present. We report that vRNAP-promoter recognition and activity on these templates require specific sequences and a hairpin structure on the template strand. Hairpin extrusion, induced by Mg(II) and physiological superhelical density, is essential to provide the correct DNA structure for polymerase recognition, as mutant promoters that do not form hairpins show reduced in vitro activity. Therefore, a supercoil-induced DNA structural transition regulates N4 vRNAP transcription. Eco SSB activates transcription at physiological superhelical densities by stabilizing the template-strand hairpin. Specific sequences at the promoters are conserved to provide proper contacts for vRNAP, to support hairpin extrusion, or both. We propose a model for in vivo utilization of the vRNAP promoters, and discuss the roles of DNA supercoiling and Eco SSB in promoter activation. PMID- 9732276 TI - Amino acid-amino acid contacts at the cooperativity interface of the bacteriophage lambda and P22 repressors. AB - The bacteriophage lambda repressor and its relatives bind cooperatively to adjacent as well as artificially separated operator sites. This cooperativity is mediated by a protein-protein interaction between the DNA-bound dimers. Here we use a genetic approach to identify two pairs of amino acids that interact at the dimer-dimer interface. One of these pairs is nonconserved in the aligned sequences of the lambda and P22 repressors; we show that a lambda repressor variant bearing the P22 residues at these two positions interacts specifically with the P22 repressor. The other pair consists of a conserved ion pair; we reverse the charges at these two positions and demonstrate that, whereas the individual substitutions abolish the interaction of the DNA-bound dimers, these changes in combination restore the interaction of both lambdacI and P22c2 dimers. PMID- 9732277 TI - Communication between Hin recombinase and Fis regulatory subunits during coordinate activation of Hin-catalyzed site-specific DNA inversion. AB - The Hin DNA invertase becomes catalytically activated when assembled in an invertasome complex containing two Fis dimers bound to an enhancer segment. The region of Fis responsible for transactivation of Hin contains a mobile beta hairpin arm that extends from each dimer subunit. We show here that whereas both Fis dimers must be capable of activating Hin, Fis heterodimers that have only one functional activating beta-arm are sufficient to form catalytically competent invertasomes. Analysis of homodimer and heterodimer mixes of different Hin mutants suggests that Fis must activate each subunit of the two Hin dimers that participate in catalysis. These experiments also indicate that all four Hin subunits must be coordinately activated prior to initiation of the first chemical step of the reaction and that the process of activation is independent of the catalytic steps of recombination. We propose a molecular model for the invertasome structure that is consistent with current information on protein-DNA structures and the topology of the DNA strands within the recombination complex. In this model, a single Fis activation arm could contact amino acids from both Hin subunits at the dimer interface to induce a conformational change that coordinately positions the active sites close to the scissile phosphodiester bonds. PMID- 9732278 TI - Dynamics of the genome during early Xenopus laevis development: karyomeres as independent units of replication. AB - During Xenopus laevis early development, the genome is replicated in less than 15 min every 30 min. We show that during this period, DNA replication proceeds in an atypical manner. Chromosomes become surrounded by a nuclear membrane lamina forming micronuclei or karyomeres. This genomic organization permits that prereplication centers gather on condensed chromosomes during anaphase and that DNA replication initiates autonomously in karyomeres at early telophase before nuclear reconstruction and mitosis completion. The formation of karyomeres is not dependent on DNA replication but requires mitotic spindle formation and the normal segregation of chromosomes. Thus, during early development, chromosomes behave as structurally and functionally independent units. The formation of a nuclear envelope around each chromosome provides an in vivo validation of its role in regulating initiation of DNA replication, enabling the rate of replication to accelerate and S phase to overlap M phase without illegitimate reinitiation. The abrupt disappearance of this atypical organization within one cell cycle after thirteen divisions defines a novel developmental transition at the blastula stage, which may affect both the replication and the transcription programs of development. PMID- 9732279 TI - Presence of pre-rRNAs before activation of polymerase I transcription in the building process of nucleoli during early development of Xenopus laevis. AB - During the early development of Xenopus laevis, we followed in individual nuclei the formation of a nucleolus by examining simultaneously its structural organization and its transcriptional competence. Three distinct situations were encountered with different frequencies during development. During the first period of general transcriptional quiescence, the transcription factor UBF of maternal origin, was present in most nuclei at the ribosomal gene loci. In contrast, fibrillarin, a major protein of the processing machinery, was found in multiple prenucleolar bodies (PNBs) whereas nucleolin was dispersed largely in the nucleoplasm. During the second period, for most nuclei these PNBs had fused into two domains where nucleolin concentrated, generating a structure with most features expected from a transcriptionally competent nucleolus. However, RNA polymerase I-dependent transcription was not detected using run-on in situ assays whereas unprocessed ribosomal RNAs were observed. These RNAs were found to derive from a maternal pool. Later, during a third period, an increasing fraction of the nuclei presented RNA polymerase I-dependent transcription. Thus, the structural organization of the nucleolus preceded its transcriptional competence. We conclude that during the early development of X. laevis, the organization of a defined nucleolar structure, is not associated with the transcription process per se but rather with the presence of unprocessed ribosomal RNAs. PMID- 9732280 TI - The hrp23 protein in the balbiani ring pre-mRNP particles is released just before or at the binding of the particles to the nuclear pore complex. AB - Balbiani ring (BR) pre-mRNP particles reside in the nuclei of salivary glands of the dipteran Chironomus tentans and carry the message for giant-sized salivary proteins. In the present study, we identify and characterize a new protein component in the BR ribonucleoprotein (RNP) particles, designated hrp23. The protein with a molecular mass of 20 kD has a single RNA-binding domain and a glycine-arginine-serine-rich auxiliary domain. As shown by immunoelectron microscopy, the hrp23 protein is added to the BR transcript concomitant with transcription, is still present in the BR particles in the nucleoplasm, but is absent from the BR particles that are bound to the nuclear pore complex or are translocating through the central channel of the complex. Thus, hrp23 is released just before or at the binding of the particles to the nuclear pore complex. It is noted that hrp23 behaves differently from two other BR RNP proteins earlier studied: hrp36 and hrp45. These proteins both reach the nuclear pore complex, and hrp36 even accompanies the RNA into the cytoplasm. It is concluded that each BR RNA-binding protein seems to have a specific flow pattern, probably related to the particular role of the protein in gene expression. PMID- 9732281 TI - Nup154, a new Drosophila gene essential for male and female gametogenesis is related to the nup155 vertebrate nucleoporin gene. AB - The Nup154 gene of Drosophila encodes a protein showing similarity with known nucleoporins: rat Nup155 and yeast Nup170 and Nup157. Hypomorphic mutant alleles of Nup154 affected female and male fertility, allowing investigation of the gene function in various steps of oogenesis and spermatogenesis. Nup154 was required in testes for cyst formation, control of spermatocyte proliferation and meiotic progression. In ovaries, Nup154 was essential for egg chamber development and oocyte growth. In both the male and female germ line, as well as in several other cell types, the Nup154 protein was detected at the nuclear membrane, but was also present inside the nucleus. Intranuclear localization has not previously been described for rat Nup155 or yeast Nup170 and Nup157. In mutant egg chambers the Nup154 protein accumulated in the cytoplasm, while it was only barely detected at the nuclear envelopes. FG repeats containing nucleoporins detected with mAb414 antibody were also mislocalized to a certain extent in Nup154 mutant alleles. This suggests that Nup154 could be required for localizing other nucleoporins within the nuclear pore complex, as previously demonstrated for the yeast Nup170. On the other hand, no evident defects in lamin localization were observed, indicating that Nup155 mutations did not affect the overall integrity of the nuclear envelope. However, ultrastructural analyses revealed that in mutant cells the morphology of the nuclear envelope was altered near the nuclear pore complexes. Finally, the multiplicity of phenotypes observed in Nup154 mutant alleles suggests that this gene plays a crucial role in cell physiology. PMID- 9732282 TI - Transport of axl2p depends on erv14p, an ER-vesicle protein related to the Drosophila cornichon gene product. AB - COPII-coated ER-derived transport vesicles from Saccharomyces cerevisiae contain a distinct set of membrane-bound polypeptides. One of these polypeptides, termed Erv14p (ER-vesicle protein of 14 kD), corresponds to an open reading frame on yeast chromosome VII that is predicted to encode an integral membrane protein and shares sequence identity with the Drosophila cornichon gene product. Experiments with an epitope-tagged version of Erv14p indicate that this protein localizes to the ER and is selectively packaged into COPII-coated vesicles. Haploid cells that lack Erv14p are viable but display a modest defect in bud site selection because a transmembrane secretory protein, Axl2p, is not efficiently delivered to the cell surface. Axl2p is required for selection of axial growth sites and normally localizes to nascent bud tips or the mother bud neck. In erv14Delta strains, Axl2p accumulates in the ER while other secretory proteins are transported at wild-type rates. We propose that Erv14p is required for the export of specific secretory cargo from the ER. The polarity defect of erv14Delta yeast cells is reminiscent of cornichon mutants, in which egg chambers fail to establish proper asymmetry during early stages of oogenesis. These results suggest an unforeseen conservation in mechanisms producing cell polarity shared between yeast and Drosophila. PMID- 9732283 TI - Degradation of misfolded endoplasmic reticulum glycoproteins in Saccharomyces cerevisiae is determined by a specific oligosaccharide structure. AB - In Saccharomyces cerevisiae, transfer of N-linked oligosaccharides is immediately followed by trimming of ER-localized glycosidases. We analyzed the influence of specific oligosaccharide structures for degradation of misfolded carboxypeptidase Y (CPY). By studying the trimming reactions in vivo, we found that removal of the terminal alpha1,2 glucose and the first alpha1,3 glucose by glucosidase I and glucosidase II respectively, occurred rapidly, whereas mannose cleavage by mannosidase I was slow. Transport and maturation of correctly folded CPY was not dependent on oligosaccharide structure. However, degradation of misfolded CPY was dependent on specific trimming steps. Degradation of misfolded CPY with N-linked oligosaccharides containing glucose residues was less efficient compared with misfolded CPY bearing the correctly trimmed Man8GlcNAc2 oligosaccharide. Reduced rate of degradation was mainly observed for misfolded CPY bearing Man6GlcNAc2, Man7GlcNAc2 and Man9GlcNAc2 oligosaccharides, whereas Man8GlcNAc2 and, to a lesser extent, Man5GlcNAc2 oligosaccharides supported degradation. These results suggest a role for the Man8GlcNAc2 oligosaccharide in the degradation process. They may indicate the presence of a Man8GlcNAc2-binding lectin involved in targeting of misfolded glycoproteins to degradation in S. cerevisiae. PMID- 9732284 TI - Ca2+ homeostasis in the agonist-sensitive internal store: functional interactions between mitochondria and the ER measured In situ in intact cells. AB - Mitochondria have a well-established capacity to detect cytoplasmic Ca2+ signals resulting from the discharge of ER Ca2+ stores. Conversely, both the buffering of released Ca2+ and ATP production by mitochondria are predicted to influence ER Ca2+ handling, but this complex exchange has been difficult to assess in situ using conventional measurement techniques. Here we have examined this interaction in single intact BHK-21 cells by monitoring intraluminal ER [Ca2+] directly using trapped fluorescent low-affinity Ca2+ indicators. Treatment with mitochondrial inhibitors (FCCP, antimycin A, oligomycin, and rotenone) dramatically prolonged the refilling of stores after release with bradykinin. This effect was largely due to inhibition of Ca2+ entry pathways at the plasma membrane, but a significant component appears to arise from reduction of SERCA-mediated Ca2+ uptake, possibly as a consequence of ATP depletions in a localized subcellular domain. The rate of bradykinin-induced Ca2+ release was reduced to 51% of control by FCCP. This effect was largely overcome by loading cells with BAPTA-AM, highlighting the importance of mitochondrial Ca2+ buffering in shaping the release kinetics. However, mitochondria-specific ATP production was also a significant determinant of the release dynamic. Our data emphasize the localized nature of the interaction between these organelles, and show that competent mitochondria are essential for generating explosive Ca2+ signals. PMID- 9732285 TI - The cytoplasmic tail of rhodopsin acts as a novel apical sorting signal in polarized MDCK cells. AB - All basolateral sorting signals described to date reside in the cytoplasmic domain of proteins, whereas apical targeting motifs have been found to be lumenal. In this report, we demonstrate that wild-type rhodopsin is targeted to the apical plasma membrane via the TGN upon expression in polarized epithelial MDCK cells. Truncated rhodopsin with a deletion of 32 COOH-terminal residues shows a nonpolar steady-state distribution. Addition of the COOH-terminal 39 residues of rhodopsin redirects the basolateral membrane protein CD7 to the apical membrane. Fusion of rhodopsin's cytoplasmic tail to a cytosolic protein glutathione S-transferase (GST) also targets this fusion protein (GST-Rho39Tr) to the apical membrane. The targeting of GST-Rho39Tr requires both the terminal 39 amino acids and the palmitoylation membrane anchor signal provided by the rhodopsin sequence. The apical transport of GST-Rho39Tr can be reversibly blocked at the Golgi complex by low temperature and can be altered by brefeldin A treatment. This indicates that the membrane-associated GST-Rho39Tr protein may be sorted along a yet unidentified pathway that is similar to the secretory pathway in polarized MDCK cells. We conclude that the COOH-terminal tail of rhodopsin contains a novel cytoplasmic apical sorting determinant. This finding further indicates that cytoplasmic sorting machinery may exist in MDCK cells for some apically targeted proteins, analogous to that described for basolaterally targeted proteins. PMID- 9732286 TI - Development of approaches to improve cell survival in myoblast transfer therapy. AB - Myoblast transplantation has been extensively studied as a gene complementation approach for genetic diseases such as Duchenne Muscular Dystrophy. This approach has been found capable of delivering dystrophin, the product missing in Duchenne Muscular Dystrophy muscle, and leading to an increase of strength in the dystrophic muscle. This approach, however, has been hindered by numerous limitations, including immunological problems, and low spread and poor survival of the injected myoblasts. We have investigated whether antiinflammatory treatment and use of different populations of skeletal muscle-derived cells may circumvent the poor survival of the injected myoblasts after implantation. We have observed that different populations of muscle-derived cells can be isolated from skeletal muscle based on their desmin immunoreactivity and differentiation capacity. Moreover, these cells acted differently when injected into muscle: 95% of the injected cells in some populations died within 48 h, while others richer in desmin-positive cells survived entirely. Since pure myoblasts obtained from isolated myofibers and myoblast cell lines also displayed a poor survival rate of the injected cells, we have concluded that the differential survival of the populations of muscle-derived cells is not only attributable to their content in desmin-positive cells. We have observed that the origin of the myogenic cells may influence their survival in the injected muscle. Finally, we have observed that myoblasts genetically engineered to express an inhibitor of the inflammatory cytokine, IL-1, can improve the survival rate of the injected myoblasts. Our results suggest that selection of specific muscle-derived cell populations or the control of inflammation can be used as an approach to improve cell survival after both myoblast transplantation and the myoblast-mediated ex vivo gene transfer approach. PMID- 9732287 TI - Differential membrane localization and intermolecular associations of alpha dystrobrevin isoforms in skeletal muscle. AB - alpha-Dystrobrevin is both a dystrophin homologue and a component of the dystrophin protein complex. Alternative splicing yields five forms, of which two predominate in skeletal muscle: full-length alpha-dystrobrevin-1 (84 kD), and COOH-terminal truncated alpha-dystrobrevin-2 (65 kD). Using isoform-specific antibodies, we find that alpha-dystrobrevin-2 is localized on the sarcolemma and at the neuromuscular synapse, where, like dystrophin, it is most concentrated in the depths of the postjunctional folds. alpha-Dystrobrevin-2 preferentially copurifies with dystrophin from muscle extracts. In contrast, alpha-dystrobrevin 1 is more highly restricted to the synapse, like the dystrophin homologue utrophin, and preferentially copurifies with utrophin. In yeast two-hybrid experiments and coimmunoprecipitation of in vitro-translated proteins, alpha dystrobrevin-2 binds dystrophin, whereas alpha-dystrobrevin-1 binds both dystrophin and utrophin. alpha-Dystrobrevin-2 was lost from the nonsynaptic sarcolemma of dystrophin-deficient mdx mice, but was retained on the perisynaptic sarcolemma even in mice lacking both utrophin and dystrophin. In contrast, alpha dystrobrevin-1 remained synaptically localized in mdx and utrophin-negative muscle, but was absent in double mutants. Thus, the distinct distributions of alpha-dystrobrevin-1 and -2 can be partly explained by specific associations with utrophin and dystrophin, but other factors are also involved. These results show that alternative splicing confers distinct properties of association on the alpha dystrobrevins. PMID- 9732288 TI - Gamma-sarcoglycan deficiency leads to muscle membrane defects and apoptosis independent of dystrophin. AB - gamma-Sarcoglycan is a transmembrane, dystrophin-associated protein expressed in skeletal and cardiac muscle. The murine gamma-sarcoglycan gene was disrupted using homologous recombination. Mice lacking gamma-sarcoglycan showed pronounced dystrophic muscle changes in early life. By 20 wk of age, these mice developed cardiomyopathy and died prematurely. The loss of gamma-sarcoglycan produced secondary reduction of beta- and delta-sarcoglycan with partial retention of alpha- and epsilon-sarcoglycan, suggesting that beta-, gamma-, and delta sarcoglycan function as a unit. Importantly, mice lacking gamma-sarco- glycan showed normal dystrophin content and local- ization, demonstrating that myofiber degeneration occurred independently of dystrophin alteration. Furthermore, beta dystroglycan and laminin were left intact, implying that the dystrophin dystroglycan-laminin mechanical link was unaffected by sarcoglycan deficiency. Apoptotic myonuclei were abundant in skeletal muscle lacking gamma-sarcoglycan, suggesting that programmed cell death contributes to myofiber degeneration. Vital staining with Evans blue dye revealed that muscle lacking gamma-sarcoglycan developed membrane disruptions like those seen in dystrophin-deficient muscle. Our data demonstrate that sarcoglycan loss was sufficient, and that dystrophin loss was not necessary to cause membrane defects and apoptosis. As a common molecular feature in a variety of muscular dystrophies, sarcoglycan loss is a likely mediator of pathology. PMID- 9732289 TI - The yeast V159N actin mutant reveals roles for actin dynamics in vivo. AB - Actin with a Val 159 to Asn mutation (V159N) forms actin filaments that depolymerize slowly because of a failure to undergo a conformational change after inorganic phosphate release. Here we demonstrate that expression of this actin results in reduced actin dynamics in vivo, and we make use of this property to study the roles of rapid actin filament turnover. Yeast strains expressing the V159N mutant (act1-159) as their only source of actin have larger cortical actin patches and more actin cables than wild-type yeast. Rapid actin dynamics are not essential for cortical actin patch motility or establishment of cell polarity. However, fluid phase endocytosis is defective in act1-159 strains. act1-159 is synthetically lethal with cofilin and profilin mutants, supporting the conclusion that mutations in all of these genes impair the polymerization/ depolymerization cycle. In contrast, act1-159 partially suppresses the temperature sensitivity of a tropomyosin mutant, and the loss of cytoplasmic cables seen in fimbrin, Mdm20p, and tropomyosin null mutants, suggesting filament stabilizing functions for these actin-binding proteins. Analysis of the cables in these double-mutant cells supports a role for fimbrin in organizing cytoplasmic cables and for Mdm20p and tropomyosin in excluding cofilin from the cables. PMID- 9732290 TI - Involvement of an actomyosin contractile ring in Saccharomyces cerevisiae cytokinesis. AB - In Saccharomyces cerevisiae, the mother cell and bud are connected by a narrow neck. The mechanism by which this neck is closed during cytokinesis has been unclear. Here we report on the role of a contractile actomyosin ring in this process. Myo1p (the only type II myosin in S. cerevisiae) forms a ring at the presumptive bud site shortly before bud emergence. Myo1p ring formation depends on the septins but not on F-actin, and preexisting Myo1p rings are stable when F actin is depolymerized. The Myo1p ring remains in the mother-bud neck until the end of anaphase, when a ring of F-actin forms in association with it. The actomyosin ring then contracts to a point and disappears. In the absence of F actin, the Myo1p ring does not contract. After ring contraction, cortical actin patches congregate at the mother-bud neck, and septum formation and cell separation rapidly ensue. Strains deleted for MYO1 are viable; they fail to form the actin ring but show apparently normal congregation of actin patches at the neck. Some myo1Delta strains divide nearly as efficiently as wild type; other myo1Delta strains divide less efficiently, but it is unclear whether the primary defect is in cytokinesis, septum formation, or cell separation. Even cells lacking F-actin can divide, although in this case division is considerably delayed. Thus, the contractile actomyosin ring is not essential for cytokinesis in S. cerevisiae. In its absence, cytokinesis can still be completed by a process (possibly localized cell-wall synthesis leading to septum formation) that appears to require septin function and to be facilitated by F-actin. PMID- 9732291 TI - Control of neuronal size homeostasis by trophic factor-mediated coupling of protein degradation to protein synthesis. AB - We demonstrate that NGF couples the rate of degradation of long-lived proteins in sympathetic neurons to the rate of protein synthesis. Inhibiting protein synthesis rate by a specific percentage caused an almost equivalent percentage reduction in the degradation rate of long-lived proteins, indicating nearly 1:1 coupling between the two processes. The rate of degradation of short-lived proteins was unaffected by suppressing protein synthesis. Included in the pool of proteins that had increased half-lives when protein synthesis was inhibited were actin and tubulin. Both of these proteins, which had half-lives of several days, exhibited no degradation over a 3-d period when protein synthesis was completely suppressed. The half-lives of seven other long-lived proteins were quantified and found to increase by 84-225% when protein synthesis was completely blocked. Degradation-synthesis coupling protected cells from protein loss during periods of decreased synthesis. The rate of protein synthesis greatly decreased and coupling between degradation and synthesis was lost after removal of NGF. Uncoupling resulted in net loss of cellular protein and somatic atrophy. We propose that coupling the rate of protein degradation to that of protein synthesis is a fundamental mechanism by which neurotrophic factors maintain homeostatic control of neuronal size and perhaps growth. PMID- 9732293 TI - The RET-glial cell-derived neurotrophic factor (GDNF) pathway stimulates migration and chemoattraction of epithelial cells. AB - Embryonic development requires cell migration in response to positional cues. Yet, how groups of cells recognize and translate positional information into morphogenetic movement remains poorly understood. In the developing kidney, the ureteric bud epithelium grows from the nephric duct towards a group of posterior intermediate mesodermal cells, the metanephric mesenchyme, and induces the formation of the adult kidney. The secreted protein GDNF and its receptor RET are required for ureteric bud outgrowth and subsequent branching. However, it is unclear whether the GDNF-RET pathway regulates cell migration, proliferation, survival, or chemotaxis. In this report, we have used the MDCK renal epithelial cell line to show that activation of the RET pathway results in increased cell motility, dissociation of cell adhesion, and the migration towards a localized source of GDNF. Cellular responses to RET activation include the formation of lamellipodia, filopodia, and reorganization of the actin cytoskeleton. These data demonstrate that GDNF is a chemoattractant for RET-expressing epithelial cells and thus account for the developmental defects observed in RET and GDNF mutant mice. Furthermore, the RET-transfected MDCK cells described in this report are a promising model for delineating RET signaling pathways in the renal epithelial cell lineage. PMID- 9732292 TI - SCAR, a WASP-related protein, isolated as a suppressor of receptor defects in late Dictyostelium development. AB - G protein-coupled receptors trigger the reorganization of the actin cytoskeleton in many cell types, but the steps in this signal transduction cascade are poorly understood. During Dictyostelium development, extracellular cAMP functions as a chemoattractant and morphogenetic signal that is transduced via a family of G protein-coupled receptors, the cARs. In a strain where the cAR2 receptor gene is disrupted by homologous recombination, the developmental program arrests before tip formation. In a genetic screen for suppressors of this phenotype, a gene encoding a protein related to the Wiskott-Aldrich Syndrome protein was discovered. Loss of this protein, which we call SCAR (suppressor of cAR), restores tip formation and most later development to cAR2(-) strains, and causes a multiple-tip phenotype in a cAR2(+) strain as well as leading to the production of extremely small cells in suspension culture. SCAR-cells have reduced levels of F-actin staining during vegetative growth, and abnormal cell morphology and actin distribution during chemotaxis. Uncharacterized homologues of SCAR have also been identified in humans, mouse, Caenorhabditis elegans, and Drosophila. These data suggest that SCAR may be a conserved negative regulator of G protein-coupled signaling, and that it plays an important role in regulating the actin cytoskeleton. PMID- 9732295 TI - Novel roles for alpha3beta1 integrin as a regulator of cytoskeletal assembly and as a trans-dominant inhibitor of integrin receptor function in mouse keratinocytes. AB - Previously we found that alpha3beta1 integrin-deficient neonatal mice develop micro-blisters at the epidermal-dermal junction. These micro-blisters were associated with poor basement membrane organization. In the present study we have investigated the effect of alpha3beta1-deficiency on other keratinocyte integrins, actin-associated proteins and F-actin organization. We show that the absence of alpha3beta1 results in an increase in stress fiber formation in keratinocytes grown in culture and at the basal face of the basal keratinocytes of alpha3-null epidermis. Moreover, we see a higher concentration of actin associated proteins such as vinculin, talin, and alpha-actinin at focal contact sites in the alpha3-deficient keratinocytes. These changes in focal contact composition were not due to a change in steady-state levels of these proteins, but rather to reorganization due to alpha3beta1 deficiency. Apart from the loss of alpha3beta1 there is no change in expression of the other integrins expressed by the alpha3-null keratinocytes. However, in functional assays, alpha3beta1 deficiency allows an increase in fibronectin and collagen type IV receptor activities. Thus, our findings provide evidence for a role of alpha3beta1 in regulating stress fiber formation and as a trans-dominant inhibitor of the functions of the other integrins in mouse keratinocytes. These results have potential implications for the regulation of keratinocyte adhesion and migration during wound healing. PMID- 9732294 TI - Mode of action of interleukin-6 on mature osteoclasts. Novel interactions with extracellular Ca2+ sensing in the regulation of osteoclastic bone resorption. AB - We describe a physiologically significant mechanism through which interleukin-6 (IL-6) and a rising ambient Ca2+ interact to regulate osteoclastic bone resorption. VOXEL-based confocal microscopy of nonpermeabilized osteoclasts incubated with anti- IL-6 receptor antibodies revealed intense, strictly peripheral plasma membrane fluorescence. IL-6 receptor expression in single osteoclasts was confirmed by in situ reverse transcriptase PCR histochemistry. IL 6 (5 ng/l to 10 microg/l), but not IL-11 (10 and 100 microg/l), reversed the inhibition of osteoclastic bone resorption induced by high extracellular Ca2+ (15 mM). The IL-6 effect was abrogated by excess soluble IL-6 receptor (500 microg/l). Additionally, IL-6 (5 pg/l to 10 microg/l) inhibited cytosolic Ca2+ signals triggered by high Ca2+ or Ni2+. In separate experiments, osteoclasts incubated in 10 mM Ca2+ or on bone released more IL-6 than those in 1.25 mM Ca2+. Furthermore, IL-6 mRNA histostaining was more intense in osteoclasts in 10 or 20 mM Ca2+ than cells in 1.25 mM Ca2+. Similarly, IL-6 receptor mRNA histostaining was increased in osteoclasts incubated in 5 or 10 mM Ca2+. Thus, while high Ca2+ enhances IL-6 secretion, the released IL-6 attenuates Ca2+ sensing and reverses inhibition of resorption by Ca2+. Such an autocrine-paracrine loop may sustain osteoclastic activity in the face of an inhibitory Ca2+ level generated locally during resorption. PMID- 9732298 TI - Importance of dietary gamma-linolenic acid in human health and nutrition. AB - Considerable debate remains regarding the distinct biological activities of individual polyunsaturated fatty acids (PUFA). One of the most interesting yet controversial dietary approaches has been the possible prophylactic role of dietary gamma-linolenic acid (GLA) in treating various chronic disease states. This strategy is based on the ability of diet to modify cellular lipid composition and eicosanoid (cyclooxygenase and lipoxygenase) biosynthesis. Recent studies demonstrate that dietary GLA increases the content of its elongase product, dihomo-gamma-linolenic acid (DGLA), within cell membranes without concomitant changes in arachidonic acid (AA). Subsequently, upon stimulation, DGLA can be converted by inflammatory cells to 15-(S)-hydroxy-8,11,13 eicosatrienoic acid and prostaglandin E1. This is noteworthy because these compounds possess both anti-inflammatory and antiproliferative properties. Although an optimal feeding regimen to maximize the potential benefits of dietary GLA has not yet been determined, it is the purpose of this review to summarize the most recent research that has focused on objectively and reproducibly determining the mechanism(s) by which GLA may ameliorate health problems. PMID- 9732296 TI - ZAP-70 tyrosine kinase is required for LFA-1-dependent T cell migration. AB - The ZAP-70 tyrosine kinase is essential for T cell activation by the T cell receptor. We show that ZAP-70 is also required for migration of T cells that is dependent on the integrin LFA-1. Invasion of TAM2D2 T cell hybridoma cells into fibroblast monolayers, which is LFA-1-dependent, was blocked by overexpression of dominant-negative ZAP-70 and by piceatannol but not by herbimycin A. The Syk inhibitor piceatannol blocks the Syk homologue ZAP-70, which is expressed by TAM2D2 cells, with the same dose dependence as the inhibition of invasion. Dominant-negative ZAP-70 completely inhibited the extensive metastasis formation of TAM2D2 cells to multiple organs upon i.v. injection into mice. Migration of TAM2D2 cells through filters coated with the LFA-1 ligand ICAM-1, induced by 1 ng/ml of the chemokine SDF-1, was blocked by anti-LFA-1 mAb and also abrogated by dominant-negative ZAP-70 and piceatannol. In contrast, migration induced by 100 ng/ml SDF-1 was independent of both LFA-1 and ZAP-70. LFA-1 cross-linking induced tyrosine phosphorylation, which was blocked by dominant-negative ZAP-70 and piceatannol. We conclude that LFA-1 engagement triggers ZAP-70 activity that is essential for LFA-1-dependent migration. PMID- 9732299 TI - Adrenocortical responsiveness to adrenocorticotropic hormone is enhanced in chronically food-restricted rats. AB - Chronic food restriction (FR) of rats and mice results in moderate hyperadrenocorticism, which may play a role in activating cellular mechanisms that retard aging. Previously, we reported that the FR-induced hyperadrenocorticism is not due to an activated hypothalamo-pituitary unit. Therefore, we investigated in a series of experiments whether adrenal responsiveness to adrenocorticotropic hormone (ACTH), in vitro and in vivo, is enhanced by FR. Three mo-old male Fischer 344 rats were either given free access to food (AL rats) or restricted to 60% of food consumed by AL rats (FR rats) from 6 wk of age. They were killed by decapitation in the morning (AM) and afternoon (PM) when endogenously circulating corticosterone levels are at their nadir and peak, respectively. In vitro, adrenal glands from FR rats (1.5 mo FR) produced more corticosterone per mg at all doses of ACTH than those from AL rats in both the AM and PM (diet main effect, P < 0.001). FR (1.5 to 2.5 mo) also enhanced adrenal responsiveness to physiologic (diet main effect, P < 0.05) and superphysiologic (diet main effect, P < 0.001) levels of ACTH administered in vivo to dexamethasone-treated rats. ACTH-receptor (ACTH-R) mRNA, normalized to adrenal mass or to total RNA, was not influenced by FR (1.5 mo). However, adrenal ACTH-R mRNA, as well as adrenal mass, per unit body weight was greater in FR than in AL rats (diet main effect, P < 0.001). These results indicate that enhanced adrenocortical responsiveness to ACTH plays a major role in the hyperadrenocortical state of chronically FR rats. PMID- 9732297 TI - Endothelial cell E- and P-selectin and vascular cell adhesion molecule-1 function as signaling receptors. AB - Previous studies have shown that polymorphonuclear leukocyte (PMN) adherence to endothelial cells (EC) induces transient increases in EC cytosolic free calcium concentration ([Ca2+]i) that are required for PMN transit across the EC barrier (Huang, A.J., J.E. Manning, T. M. Bandak, M.C. Ratau, K.R. Hanser, and S.C. Silverstein. 1993. J. Cell Biol. 120:1371-1380). To determine whether stimulation of [Ca2+]i changes in EC by leukocytes was induced by the same molecules that mediate leukocyte adherence to EC, [Ca2+]i was measured in Fura2-loaded human EC monolayers. Expression of adhesion molecules by EC was induced by a pretreatment of the cells with histamine or with Escherichia coli lipopolysaccharide (LPS), and [Ca2+]i was measured in single EC after the addition of mAbs directed against the EC adhesion proteins P-selectin, E-selectin, intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), or platelet/endothelial cell adhesion molecule-1 (PECAM-1). Both anti-P- and anti-E-selectin mAb, as well as anti-VCAM-1 mAb, induced transient increases in EC [Ca2+]i that were comparable to those induced by 200 microM histamine. In contrast, no effect was obtained by mAbs directed against the endothelial ICAM-1 or PECAM-1. PMN adherence directly stimulated increases in [Ca2+]i in histamine- or LPS-treated EC. mAbs directed against leukocyte CD18 or PECAM-1, the leukocyte counter receptors for endothelial ICAM-1 and PECAM-1, respectively, did not inhibit PMN induced EC activation. In contrast, mAb directed against sialyl Lewis x (sLex), a PMN ligand for endothelial P- and E-selectin, completely inhibited EC stimulation by adherent PMN. Changes in EC [Ca2+]i were also observed after adherence of peripheral blood monocytes to EC treated with LPS for 5 or 24 h. In these experiments, the combined addition of mAbs to sLex and VLA-4, the leukocyte counter-receptor for endothelial VCAM-1, inhibited [Ca2+]i changes in the 5 h treated EC, whereas the anti-VLA-4 mAb alone was sufficient to inhibit [Ca2+]i changes in the 24 h-treated EC. Again, no inhibitory effect was observed with an anti-CD18 or anti-PECAM-1 mAb. Of note, the conditions that induced changes in EC [Ca2+]i, i.e. , mAbs directed against endothelial selectins or VCAM-1, and PMN or monocyte adhesion to EC via selectins or VCAM-1, but not via ICAM-1 or PECAM-1, also induced a rearrangement of EC cytoskeletal microfilaments from a circumferential ring to stress fibers. We conclude that, in addition to their role as adhesion receptors, endothelial selectins and VCAM-1 mediate endothelial stimulation by adhering leukocytes. PMID- 9732300 TI - Dietary menhaden and corn oils and the red blood cell membrane lipid composition and fluidity in hyper- and normocholesterolemic miniature swine. AB - Fatty acids in the diet are readily incorporated into lipids in various tissues. However, it is not clear whether all tissues have the same level of incorporation. Second, (n-6) unsaturated fatty acids increase the fluidity of membranes, but this has not been shown for (n-3) fatty acids. In this study, we measured the incorporation of (n-6) and (n-3) fatty acids into erythrocyte membrane lipids and studied their effects on the fluidity of erythrocyte membranes. One group of female miniature swine was made hypercholesterolemic by feeding the swine cholesterol and lard for 2 mo; the other group served as controls and was fed a stock diet. Both groups were then fed either corn oil or menhaden oil or a mixture of the two for 23 additional weeks. Blood was collected at 0, 2, 4, 12 and 23 wk after initialization of the experimental diets, and fatty acid composition of phospholipids was assessed. Membrane phospholipids of pigs fed menhaden oil had elevated (n-3) fatty acids (20:5 and 22:6), and lower 18:2 than those fed corn oil. There was no difference in 20:4 content. The fatty acid changes occurred as early as 2 wk after consumption of the corn oil or menhaden oil in pigs previously fed a stock diet, but it took longer in pigs previously fed lard + cholesterol, indicating residual effects of pretreatment. Menhaden oil increased anisotropy (indicating decreased fluidity) more than corn oil for the nonpolar probe diphenylhexatriene (DPH) at earlier time points, but not at 23 wk. Erythrocyte membrane fluidity was significantly related to membrane polyunsaturate content, with (n-6) fatty acids having a greater influence than (n 3) fatty acids. A comparison of the present red blood cell fatty acid compositions with brain synaptosome fatty acid compositions for the same animals showed poor correlations for some of the fatty acids. There was no significant direct relationship between docosahexaenoate (DHA) concentrations in erythrocyte membranes with DHA concentrations in brain synaptosomes from cerebellum, forebrain and caudate nucleus. PMID- 9732301 TI - Addition of guar gum and soy protein increases the efficacy of the American Heart Association (AHA) step I cholesterol-lowering diet without reducing high density lipoprotein cholesterol levels in non-human primates. AB - The aim of this study was to determine whether the addition of soy protein and guar gum to the American Heart Association (AHA) Step I diet would increase its efficacy compared with the typical "Average American Diet" (AAD) in a non-human primate model. Twenty adult female cynomolgus monkeys (Macaca fascicularis) were fed one of three diets for 6 wk. The AAD contained 36% energy from fat; the standard Step I diet contained 30% energy from fat; and the modified AHA Step I diet contained 30% energy from fat with the addition of soy protein isolate (10% of total energy) and guar gum (5.8 g/d). Plasma samples were collected from food deprived monkeys at 4, 5 and 6 wk of dietary treatment for analyses of plasma total cholesterol (TC), lipoprotein cholesterol and triacylglycerol (TAG) concentrations. Plasma TC, LDL-C, HDL-C and TAG concentrations were not significantly different in wk 4, 5 and 6 within any of the diet periods; thus the three measurements were averaged. After 6 wk of dietary treatment, monkeys fed the standard Step I diet had lower plasma TC (-19%) (P < 0.05) and LDL cholesterol (LDL-C) (-24%) (P < 0.09) than when they were fed the AAD, with no effect on HDL cholesterol (HDL-C), the lipoprotein cholesterol profile or TAG. Beyond the effect of the standard Step I diet, the modified AHA Step I diet further reduced plasma TC and LDL-C (-24% and -40%) (P < 0. 05) and the TC/HDL-C and LDL-C/HDL-C ratios (-37% and -52%) (P < 0. 05) with no significant changes in plasma HDL-C or TAG. The primary conclusions of this study are that the efficacy of the AHA Step I cholesterol-lowering diet can be increased with the addition of soy protein and guar gum and provide a more favorable lipoprotein cholesterol profile. Whether the cholesterol-lowering effect is the result of soy protein or guar gum or a synergistic effect of both remains to be determined. PMID- 9732302 TI - Dietary soluble fiber lowers plasma LDL cholesterol concentrations by altering lipoprotein metabolism in female guinea pigs. AB - This experiment was designed to evaluate the effects of pectin (PE), guar gum (GG) and psyllium (PSY) intake on VLDL and LDL metabolism in female guinea pigs fed high dietary cholesterol. Guinea pigs were fed a 15 g/100 g fat diet containing 0.25 g/100 g cholesterol with 12.5 g/100 g PE, 12.5 g/100 g GG, 7.5 g/100 g PSY or 12.5 g/100 g cellulose (control diet) for 4 wk. Plasma cholesterol concentrations were 29, 43 and 39% lower in guinea pigs fed PE, GG or PSY, respectively, compared with the control group (P < 0.0001). Plasma apolipoprotein (apo) B concentrations were 16-22% lower in the groups fed soluble fiber compared with the control group (P < 0.01). In contrast, hepatic cholesterol and triglyceride concentrations were not different among the PE, GG, PSY and control groups. No differences in triacylglycerol (TAG) or apo B secretion rates, measured by blocking VLDL catabolism by triton (WR 1339) injection, were observed, whereas plasma LDL apo B fractional catabolic rates (FCR), determined by injection of radiolabeled LDL, were higher in guinea pigs fed GG or PSY than in those from the control group. All sources of dietary soluble fiber reduced LDL apo B flux (P < 0.05). These results suggest that the mechanisms of plasma LDL cholesterol lowering by dietary soluble fiber are distinctive for each fiber source and result in specific alterations in lipoprotein metabolism in female guinea pigs. Differences between male and female guinea pigs in response to these diets are discussed. PMID- 9732304 TI - Cigarette smoking is associated with unhealthy patterns of nutrient intake: a meta-analysis. AB - The aim of this investigation was to assess the relationship between smoking status and nutrient intakes using a meta-analysis. Publications in English were sought through a Medline search using the following key words: food habits, eating, feeding behavior, diet, food, nutrition, nutritional status or assessment, tobacco use disorder, tobacco, nicotine and smoking. Scanning relevant reference lists of articles and hand searching completed the data collection. No attempt was made to search for unpublished results. Paper selection was based on nutritional surveys including comparisons of smokers with nonsmokers. Fifty-one published nutritional surveys from 15 different countries with 47,250 nonsmokers and 35,870 smokers were used in the analysis. The estimates of size effects were calculated with the mean and variance values of each nutrient intake and the size of the sample. Smokers declared significantly (all P < 10(-5)) higher intakes of energy (+4.9%), total fat (+3.5%), saturated fat (+8.9%), cholesterol (+10.8%) and alcohol (+77.5%) and lower intakes of polyunsaturated fat (-6.5%), fiber (-12.4%), vitamin C (-16.5%), vitamin E ( 10.8%) and beta-carotene (-11.8%) than nonsmokers. Protein and carbohydrate intakes did not differ between smokers and nonsmokers. There was no evidence of heterogeneity among studies. In conclusion, the nutrient intakes of smokers differ substantially from those of nonsmokers. Some of these differences may exacerbate the deleterious effects of smoke components on cancer and coronary heart disease risk. PMID- 9732303 TI - Long-term fructose consumption accelerates glycation and several age-related variables in male rats. AB - Fructose intake has increased steadily during the past two decades. Fructose, like other reducing sugars, can react with proteins through the Maillard reaction (glycation), which may account for several complications of diabetes mellitus and accelerating aging. In this study, we evaluated the effect of fructose intake on some age-related variables. Rats were fed for 1 y a commercial nonpurified diet, and had free access to water or 250 g/L solutions of fructose, glucose or sucrose. Early glycation products were evaluated by blood glycated hemoglobin and fructosamine concentrations. Lipid peroxidation was estimated by urine thiobarbituric reactive substances. Skin collagen crosslinking was evaluated by solubilization in natural salt or diluted acetic acid solutions, and by the ratio between beta- and alpha-collagen chains. Advanced glycation end products were evaluated by collagen-linked fluorescence in bones. The ratio between type-III and type-I collagens served as an aging variable and was measured in denatured skin collagen. The tested sugars had no effect on plasma glucose concentrations. Blood fructose, cholesterol, fructosamine and glycated hemoglobin levels, and urine lipid peroxidation products were significantly higher in fructose-fed rats compared with the other sugar-fed and control rats. Acid-soluble collagen and the type-III to type-I ratio were significantly lower, whereas insoluble collagen, the beta to alpha ratio and collagen-bound fluorescence at 335/385 nm (excitation/emission) were significantly higher in fructose-fed rats than in the other groups. The data suggest that long-term fructose consumption induces adverse effects on aging; further studies are required to clarify the precise role of fructose in the aging process. PMID- 9732305 TI - Vitamin A or beta-carotene supplementation reduces but does not eliminate maternal night blindness in Nepal. AB - We investigated the effect of supplementing women weekly with 7000 microg retinol equivalents as preformed vitamin A or beta-carotene vs. a placebo, on the incidence of night blindness during pregnancy and the postpartum period in the rural plains of Nepal. Over a period of approximately 3 y, approximately 29,000 women of child-bearing age, living in 171 wards that were randomized to one of the three supplements, contributed 9932 first pregnancies. A prospective, weekly surveillance identified night blindness in pregnant women, verified further by detailed questioning about nighttime vision. After delivery, women were also interviewed at approximately 3 and approximately 6 mo postpartum to elicit a night blindness history over the preceding 3 mo. Vitamin A supplementation reduced the incidence of night blindness during pregnancy from 10.7% among controls to 6.7% (relative risk 0.62, 95% confidence interval: 0.45-0.85). beta Carotene supplementation had less of an effect (0. 83, 0.63-1.11). Among women who took >95% of their vitamin A supplements during pregnancy, incidence of verified night blindness was reduced by 67%. Incidence (per 100 person-years) of night blindness during the first 3 mo postpartum was 11.3 in the control, 4.3 in the vitamin A and 8.7 in the beta-carotene groups, yielding corresponding relative risks of 0.38 (0.26-0.55) and 0.77 (0.57-1. 04). In the second 3 mo postpartum, both vitamin A and beta-carotene reduced night blindness by approximately 50%. Vitamin A intakes approaching a recommended amount for pregnancy markedly reduced but did not eliminate night blindness in Nepali women. Greater intakes of vitamin A than provided and/or other nutrients may be needed to prevent maternal night blindness in rural South Asia. PMID- 9732306 TI - Obesity in Latin American women and children. AB - National surveys conducted since 1982 were used to assess maternal and child obesity in Latin American and Caribbean countries and in U. S. residents of Mexican descent. Obesity in women, a body mass index (BMI) >/=30 kg/m2, was 3% in Haiti, 8-10% in eight Latin American countries and 29% in Mexican Americans. Median BMI for Latin American women were near or above the 50th percentile of the general U.S. population; values exceeded the 75th percentile in the case of Mexican Americans. The prevalence of overweight (>1 SD above mean weight-for height) in children 1-5 y of age ranged from 6% in Haiti to 24% in Peru among 13 countries. Overweight occurred in 24% of Mexican-American children. Prevalences of overweight in children and of obesity in women were greater in urban areas and in households of higher socioeconomic status. Overweight in children increased with higher maternal education; however, in some countries, obesity in women decreased with higher education. No general pattern of change over time was observed in eight countries in overweight in children. Obesity in women increased in the three countries with such data and in Mexican-American women and children. There was a tendency for greater national incomes to be associated with greater obesity levels in women and with lower levels of stunting in children. Levels of obesity in the region indicate a public health concern, particularly among women, considering that studies have identified mortality and morbidity risks associated with obesity in adults. PMID- 9732307 TI - Development and regulation of calcium metabolism in healthy girls. AB - The major components of calcium metabolism, as evaluated by a dual-tracer stable isotope method, were determined in 100 studies of 68 healthy girls, aged 5-18 y and analyzed from a developmental and regulatory viewpoint. Bone calcium deposition and removal rates were closely correlated with the size of the exchangeable bone calcium compartment. All three quantities, as well as intestinal calcium absorption, peaked at or near menarche. Both bone calcium deposition and removal rates were positively and linearly correlated with calcium absorption. However, in this correlation, because bone calcium deposition increased 70% faster than calcium absorption, most of the increase in the bone calcium compartment and its turnover must have occurred in response to something other than intestinal calcium input; presumably this occurred in response to developmental signals. Nevertheless, the constancy of the serum calcium in the face of a large intestinal calcium input and the modest way in which excretion overcame the calcium load in this population point to the importance of the exchangeable bone calcium compartment, in dynamic equilibrium with the bone mineral, as the site at which most of the load is taken up. In this population of girls, as in older women, this increase in the skeletal calcium balance resulted from a decrease in the bone calcium removal rate that was greater than the corresponding increase in the bone calcium deposition rate. PMID- 9732308 TI - D-Tagatose, a stereoisomer of D-fructose, increases hydrogen production in humans without affecting 24-hour energy expenditure or respiratory exchange ratio. AB - In growth studies on rats, the ketohexose D-tagatose has been shown to contribute no net metabolizable energy, and a pronounced thermic effect of the sugar has been suggested to account for the absence of energy. In a double-blind and balanced cross-over design, we measured 24-h energy expenditure in eight normal weight humans in a respiration chamber during the consumption of 30 g D-tagatose or 30 g sucrose/d. Metabolic measurements were performed before and after a 2-wk adaptation period with a 30-g daily intake of the test sugar. Total 24-h energy expenditure and hour-by-hour profile were unaffected by the test sugar. The nonprotein respiratory exchange ratio (RERnp) was similar during consumption of D tagatose and sucrose. However, the effect on RERnp due to CO2 produced by fermentation of D-tagatose could not be quantified in this study. A significant increase in 24-h H2 production (35%) during D-tagatose administration suggests a substantial malabsorption of the sugar. We found no effects of the 2-wk adaptation period on the measured gas exchange variables. Significantly lower fasting plasma insulin and triglyceride concentrations were observed during D tagatose administration compared with the sucrose period. No effects of D tagatose on body weight and composition were seen, but the perception of fullness 2.5 h after the sugar load was greater with D-tagatose. In conclusion, this study does not suggest a pronounced thermic effect of D-tagatose, and other mechanisms seem to be required to explain its lack of net energy. PMID- 9732309 TI - Opposite fluxes of glutamine and alanine in the splanchnic area are an efficient mechanism for nitrogen sparing in rats. AB - Glutamine release by the liver constitutes a process of nitrogen salvage through the recycling of a part of the nitrogen, which prevents irreversible nitrogen losses as urea. The aim of this work was to study the nitrogen cycling in the splanchnic bed under different nutritional conditions: fed state, postabsorptive state (16 h food deprivation) or prolonged starvation (24 or 40 h). Rats were adapted to a 15% casein diet for 15 d and then sampled. The digestive, hepatic and splanchnic balances of glucose, lactate, ketone bodies, urea and amino acids were determined. There was a net release of lactate and alanine by the digestive tract, due to the high rate of glycolysis and glutaminolysis. During prolonged starvation, ketone bodies became major energy fuel for the intestine. In fed rats, there was a net uptake of most amino acids by the liver, except for glutamine and glutamate. Urea, glutamine and glutamate released represented 33, 24 and 6% of total nitrogen taken up by the liver, respectively. In postabsorptive rats, compared with fed rats, there was a significant reduction of ureagenesis, and glutamine became the major form of nitrogen released by the liver. In fact, nitrogen cycling in the form of glutamine or glutamate in the liver may be interpreted as a nitrogen salvage process, rather than as an acid base control process. In the splanchnic area, in parallel with a highly active cycling of glucose as lactate, there exists a nitrogen cycling involving opposite fluxes of glutamine and alanine. PMID- 9732310 TI - Dietary flavonoids reduce lipid peroxidation in rats fed polyunsaturated or monounsaturated fat diets. AB - We investigated the influence of dietary flavonoids on alpha-tocopherol status and LDL peroxidation in rats fed diets enriched in either polyunsaturated fatty acids (PUFA) or monounsaturated fatty acids (MUFA). Diets equalized for alpha tocopherol concentrations were or were not supplemented with 8 g/kg diet of flavonoids (quercetin + catechin, 2:1). After 4 wk of feeding, plasma lipid concentrations were lower in rats fed PUFA than in those fed MUFA with a significant correlation between plasma alpha-tocopherol and cholesterol concentrations, r = 0.94, P < 0. 0001). Dietary lipids influenced the fatty acid composition of VLDL + LDL more than that of HDL or microsomes. The resistance of VLDL + LDL to copper-induced oxidation was higher in rats fed MUFA than in those fed PUFA as assessed by the lower production of conjugated dienes and thiobarbituric acid reactive substances (TBARS) and by the >100% longer lag time for dienes production. (P < 0.0001). Dietary flavonoids significantly reduced by 22% the amounts of dienes produced during 12 h of oxidation in rats fed diets rich in PUFA and lengthened lag time 43% in those fed MUFA. Microsomes of rats fed MUFA produced approximately 50% less TBARS than those of rats fed PUFA (P < 0.0001) and they contained more alpha-tocopherol in rats fed MUFA than in those fed PUFA with higher values (P < 0. 0001) in both groups supplemented with flavonoids (P < 0.0001). Our findings suggest that the intake of dietary flavonoids is beneficial not only when diets are rich in PUFA but also when they are rich in MUFA. It seems likely that these substances contribute to the antioxidant defense and reduce the consumption of alpha-tocopherol in both lipoproteins and membranes. PMID- 9732311 TI - Rates of lysine catabolism are inversely related to rates of protein synthesis when measured concurrently in adult female rats induced to grow at different rates. AB - To test the effect of changes in the rate of protein synthesis on amino acid oxidation, both were studied concurrently in individual 200-g female Sprague Dawley rats. In a growth trial (Experiment 1), recombinant bovine somatotropin (rbST) was injected subcutaneously (0, 2 or 12 mg/d) over 6 d (n = 4 rats per rbST level). Weight gain increased with rbST level (P < 0.01); 1.96 +/- 0.8, 4.24 +/- 0.8 and 8.67 +/- 0.8 g/d, respectively. After treatment with rbST (0 or 12 mg/d) for 4 d (Experiment 2), rats were injected via a tail vein catheter with valine (400 mmol, 4.07 mBq L-[3,4(n)-3H]valine) at 0, 4, 10, 13 or 16 h after the daily rbST injection and killed 20 min later. This flooding dose was 5 to 6 times, not 10 times, the free pool as hoped. Protein synthesis in rbST-treated rats increased 46% in muscle (P < 0.001) and 36% in liver (P < 0.01). The ks was unaltered with time after rbST injection (0-16 h, P > 0.05). When 600 mmol valine (4.4 mBq L-[3,4(n)-3H]valine) was used in Experiment 3, specific activity (SA) of free valine was constant over 20 min and was 94 +/- 4% of that injected. Finally, in Experiment 4, protein synthesis and amino acid oxidation rates measured in the same rat revealed a 35% increase (P < 0.01) in protein synthesis in hind leg muscle and a 29% increase in liver (P < 0.05) from rbST-injected (12 mg/d) rats (n = 6). Lysine oxidation was estimated by continuous (12 h) infusion of L-[1 14C]lysine via the opposite tail vein catheter. Expired CO2 was collected over 20 min intervals and SA at plateau was estimated by fitting an exponential model. Lysine oxidation was reduced (P < 0.05) by 44% in rbST-treated rats. The idea that an increase in protein synthesis results in decreased amino acid oxidation remains tenable. PMID- 9732312 TI - Plasma and hepatic cholesterol levels and fecal neutral sterol excretion are altered in hamsters fed straw mushroom diets. AB - The effect of the fruiting body and mycelium of Volvariella volvacea (straw mushroom) on the concentrations of plasma lipids, liver cholesterol, fecal neutral sterol and bile acid excretions was investigated in male Golden Syrian hamsters. The hamsters were fed a purified hypercholesterolemic diet (0.1% cholesterol, 10% fat) for 4 wk to elevate plasma lipid concentrations. Twelve hamsters with elevated plasma total cholesterol were randomly assigned to each treatment group: control (5% cellulose), mushroom fruiting body (5%) and mushroom mycelium (5%). After 4 wk of mushroom diet consumption, the plasma total cholesterol, HDL cholesterol, and combined VLDL + LDL cholesterol concentrations (mmol/L) were significantly lower than control in the group fed the fruiting body diet (40, 38 and 43%, respectively) (P < 0.05). The liver cholesterol levels were significantly lower in both the mushroom fruiting body- and the mycelium-fed groups (28 and 21% in terms of concentration; 39 and 30% in terms of total content, respectively) (P < 0.05) than that in the control group. Fecal neutral sterol excretion in the mushroom fruiting body- and mycelium-fed groups was significantly higher (81 and 74%, respectively) (P < 0.05) than that in the control group. Although no significant differences (P > 0.05) in the excretion of fecal bile acids were observed among groups fed the mushroom diets and the control diet, the mushroom fruiting body diet-fed hamsters apparently had less bacterial degradation of cholic acid as indicated by a significantly greater proportion (P < 0.05) of fecal cholic acid than in controls. They also had a significantly lower proportion of fecal deoxycholic acid (P < 0.05). This study suggests that the fruiting body of the straw mushroom lowers elevated plasma cholesterol in hypercholesterolemic hamsters, whereas the mycelium does not. PMID- 9732313 TI - Dietary amino acids are the preferential source of hepatic protein synthesis in piglets. AB - To investigate the utilization of dietary amino acids for hepatic protein synthesis, seven female pigs ( 28 d old, 7.5 kg) were implanted with catheters in a carotid artery, the jugular and portal veins, and the stomach. A portal flow probe was also implanted. The pigs were fed a high protein diet once hourly and infused intragastrically with [U-13C]algal protein for 6 h. Amino acid labeling was measured in arterial and portal blood, in the hepatic free and protein-bound pools and in apolipoprotein B-100 (apoB-100), albumin and fibrinogen. The isotopic enrichments of apoB-100-bound [U-13C]threonine, leucine, lysine and phenylalanine were 33, 100, 194 and 230% higher than those of their respective hepatic free amino acid pools (P < 0.01). Using the labeling of apoB-100 to estimate that of the protein synthetic precursor, the fractional rate of hepatic protein synthesis was 42 +/- 2%/d. Between 5 and 8% of the dietary tracer amino acids was used for hepatic protein synthesis. In contrast to the small intestinal mucosa, in which the majority of the metabolized amino acids were apparently catabolized, protein synthesis utilized from 48% (threonine) to 90% (lysine) of the hepatic uptake of tracer amino acids. It appears that hepatic protein synthesis consumes nutritionally significant quantities of dietary essential amino acids in first pass and that extracellular, especially portal, essential amino acids are channeled to hepatic protein synthesis in the fed state. PMID- 9732314 TI - Duodenal infusions of palmitic, stearic or oleic acids differently affect mammary gland metabolism of fatty acids in lactating dairy cows. AB - The effect of dietary lipids on the fatty acid (FA) profile of cows' milk fat is mainly dependent on digestive processes and mammary gland uptake and metabolism of FA. The objective of this study was to determine the separate effects of high arterial concentrations of 16:0, 18:0 and cis-18:1(n-9) on uptake, synthesis and 18:0 desaturation rate in the mammary gland of lactating dairy cows, via arterio venous differences and mammary gland balance of FA. In a 4 x 4 Latin square, four lactating Holstein cows with cannula in the proximal duodenum were infused duodenally with a mixture providing daily 0 (C treatment) or 500 g FA with mainly 16:0 (P treatment), 18:0 (S treatment) or cis-18:1(n-9) (O treatment). Significantly higher arterial concentrations of infused FA in arterial plasma nonesterified FA and triglycerides (NETGFA) were observed with P and O treatments, but the effect of the S treatment was much lower. Arterio-venous differences of NETGFA increased with arterial concentrations. The number of synthesized FA in the mammary gland was not significantly affected by duodenal infusion of FA. Mean chain length was significantly reduced by P and O treatments, suggesting an effect of mammary gland uptake of long-chain FA on the termination process of mammary gland synthesis of FA. Across all treatments, 4:0 mammary gland balance increased linearly (r = 0.67, P = 0.004) with mammary gland FA uptake. Mammary gland desaturation of 18:0 to cis-18:1(n-9) averaged 52% and was not significantly affected by treatments, but was reduced by trans-18:1 mammary gland uptake. Uptake, synthesis and desaturation of FA by the mammary gland of dairy cows are affected by arterial concentrations of 16:0, 18:0 and cis 18:1(n-9). PMID- 9732315 TI - The quantity of dietary protein affects brain protein synthesis rate in aged rats. AB - The purpose of this study was to determine whether the quantity of dietary protein affects the rate of brain protein synthesis in aged rats. Experiments were conducted on three groups of 30-wk-old rats fed diets containing 0, 5 or 20 g casein/100 g for 10 d. The fractional rates of protein synthesis in brain, liver and kidney declined with a decrease in quantity of dietary protein. In brain, liver and kidney, RNA activity [g protein synthesized/(g RNA.d)] was significantly correlated with the fractional rate of protein synthesis. The RNA concentration (mg RNA/g protein) was not related to the fractional rate of protein synthesis in any organ. The results suggest that the rate of protein synthesis in the brain declines with a decrease in quantity of dietary protein in aged rats, and that RNA activity is at least partly related to the fractional rate of brain protein synthesis. PMID- 9732316 TI - Alpha-linolenic acid deficiency modifies distractibility but not anxiety and locomotion in rats during aging. AB - In rodents, chronic dietary alpha-linolenic acid deficiency decreases learning and memory and alters dopaminergic and serotoninergic neurotransmission. However, these two neurotransmitter systems are related mainly to attention, emotion and locomotion. Therefore, we decided to investigate the effects of dietary alpha linolenic acid deficiency in rats tested with animal models of distractibility (the distractometer procedure), anxiety (the elevated plus maze) and ambulatory activity (a circular corridor). Moreover, because these neurochemical modifications persist during aging, we decided to study the effects of aging on these behaviors by using rats aged 2, 6, 12 and 24 mo. An age-related decline in distractibility was observed that was accelerated by linolenic acid deficiency. Indeed, an age-related reduction in distractibility was found in so far as distraction time was reduced at the age of 12 mo in controls and at the age of 24 mo in deficient groups compared with 2-mo-old rats. Moreover, distraction time was significantly lower in 6- and 24-mo-old rats fed a deficient diet compared with age-matched controls. Anxiety was not modified by diet or age. Finally, a parallel decrease in locomotion was exhibited by rats fed both diets between 6 and 12 mo of age. Locomotion was not modified by diet. These results show that dietary alpha-linolenic deficiency alters behavior in a very specific way; distractibility is modified by diet, whereas anxiety and locomotion are not, suggesting that particular brain areas may be altered. PMID- 9732317 TI - An evaluation of dual-energy X-Ray absorptiometry and underwater weighing to estimate body composition by means of carcass analysis in piglets. AB - To evaluate the use of dual-energy X-ray absorptiometry (DXA) and underwater weighing (UWW) for body-composition measurements, the carcasses of eight piglets (12-wk old, 15-22 kg in weight) were dissected into muscle, fat and bone. Thereafter, the components were homogenized and chemically analyzed for fat and bone mineral mass. Body components as measured by DXA correlated closely to the carcass analysis (r = 0.90-1.0). However, DXA still overestimated significantly the bone mineral mass, lean mass and total weight, and underestimated fat mass. The reproducibility of measurements, expressed as the CV for fat mass was 13.5%, whereas for total weight, lean mass and bone mineral mass, the CV was 0.74-1.9%. Fat mass was overestimated by UWW using the equations of Siri or Kraybill (r = 0. 77), but not by the equation of Lohman et al. (r = 0.69). The difference between the estimation of fat by chemical analysis and estimations by DXA and UWW was significantly affected by the amount of water in lean mass and fat-free mass. PMID- 9732318 TI - Evaluation of dual-energy X-Ray absorptiometry for body-composition assessment in rats. AB - Recent developments in dual-energy X-ray absorptiometry (DXA) have rendered feasible the determination of whole-body composition in small laboratory animals by directly measuring fat, fat-free and mineral bone masses. Our aim was to evaluate this technique by cross-calibrating the DXA method with the carcass chemical analysis in a heterogeneous population of nondiabetic Wistar and diabetic GK rats (21 animals were used for precision error and reproducibility determinations and 26 were used for accuracy studies). We report that this technique is optimized for weights >200 g. The respective CV for lean mass, fat mass and percentage of fat mass determined in short-term or transversal studies was 1.1 +/- 0.1, 3.0 +/- 1.3 and 3. 1 +/- 0.4% (mean +/- SD) respectively. Further, this technique is valid for rats weighing from 130 to 200 g by using three successive scans. In longitudinal studies, daily calibrations significantly increased the percentage of fat mass CV to 6.6 +/- 3.3%, but it was significantly decreased to 3.0 +/- 2.7% by the use of triplicate scans. The accuracy for DXA was excellent in reference to the chemical extraction technique (r2 = 0.95 for percentage of fat mass, P < 0.0001), using an adjustment factor of 0.75 (limits of agreement between the two methods for percentage of fat mass = -1.7-2.3%). Mimicry of longitudinal changes in body composition with intraperitoneal injections of saline solution demonstrated a satisfactory detection of body component changes (=300 mg/l (5% and 9%). The proportion of patients with hypoglycaemic attacks was not different between groups, but mean weight gain in the atenolol group was greater (3.4 kg v 1.6 kg). CONCLUSION: Blood pressure lowering with captopril or atenolol was similarly effective in reducing the incidence of diabetic complications. This study provided no evidence that either drug has any specific beneficial or deleterious effect, suggesting that blood pressure reduction in itself may be more important than the treatment used. PMID- 9732340 TI - Relation between iron stores and non-insulin dependent diabetes in men: case control study. PMID- 9732341 TI - Trends in smoking during pregnancy in England, 1992-7: quota sampling surveys. PMID- 9732339 TI - Cost effectiveness analysis of improved blood pressure control in hypertensive patients with type 2 diabetes: UKPDS 40. UK Prospective Diabetes Study Group. AB - OBJECTIVES: To estimate the economic efficiency of tight blood pressure control, with angiotensin converting enzyme inhibitors or beta blockers, compared with less tight control in hypertensive patients with type 2 diabetes. DESIGN: Cost effectiveness analysis incorporating within trial analysis and estimation of impact on life expectancy through use of the within trial hazards of reaching a defined clinical end point. Use of resources driven by trial protocol and use of resources in standard clinical practice were both considered. SETTING: 20 hospital based clinics in England, Scotland, and Northern Ireland. SUBJECTS: 1148 hypertensive patients with type 2 diabetes from UK prospective diabetes study randomised to tight control of blood pressure (n=758) or less tight control (n=390). MAIN OUTCOME MEASURE: Cost effectiveness ratios based on (a) use of healthcare resources associated with tight control and less tight control and treatment of complications and (b) within trial time free from diabetes related end points, and life years gained. RESULTS: Based on use of resources driven by trial protocol, the incremental cost effectiveness of tight control compared with less tight control was cost saving. Based on use of resources in standard clinical practice, incremental cost per extra year free from end points amounted to pound1049 (costs and effects discounted at 6% per year) and pound434 (costs discounted at 6% per year and effects not discounted). The incremental cost per life year gained was pound720 (costs and effects discounted at 6% per year) and pound291 (costs discounted at 6% per year and effects not discounted). CONCLUSIONS: Tight control of blood pressure in hypertensive patients with type 2 diabetes substantially reduced the cost of complications, increased the interval without complications and survival, and had a cost effectiveness ratio that compares favourably with many accepted healthcare programmes. PMID- 9732342 TI - Risk of testicular cancer with cryptorchidism and with testicular biopsy: cohort study. PMID- 9732344 TI - The cycle and its medical foes PMID- 9732343 TI - Is general practice in need of a career structure? PMID- 9732345 TI - Ductal carcinoma in situ of the breast. PMID- 9732347 TI - The golden mean PMID- 9732346 TI - Childhood Cushing's syndrome induced by betamethasone nose drops, and repeat prescriptions. PMID- 9732348 TI - Reforming the Russian health service. PMID- 9732349 TI - Late onset genetic disease: where ignorance is bliss, is it folly to inform relatives? PMID- 9732351 TI - Association between plasma plasminogen activator inhibitor-1 and survival in colorectal cancer. Measuring C reactive protein concentrations may be more useful. PMID- 9732350 TI - Screening for nuchal translucency. Measurements give parents useful information. PMID- 9732352 TI - Public concern about complaints against doctors is widespread. PMID- 9732353 TI - Trial is needed of ACE inhibitors plus beta blockers in survivors of myocardial infarction. PMID- 9732354 TI - Doctors must be trained to deal with adolescents. PMID- 9732355 TI - Interrupting the sympathetic outflow in causalgia and reflex sympathetic dystrophy. Terminology is outdated-1994 taxonomy should be used. PMID- 9732357 TI - Jonathan mann PMID- 9732356 TI - Elements of decentralisation in plans to reform NHS may prevail. PMID- 9732358 TI - The value of life PMID- 9732359 TI - Anna: too much, too young PMID- 9732360 TI - Prison to the community in one, totally unprepared, step PMID- 9732361 TI - Good news on diabetes, bad news on nose drops PMID- 9732362 TI - Tight blood pressure control reduces risks of type 2 diabetes and is cost effective PMID- 9732364 TI - British women persist in smoking during pregnancy PMID- 9732363 TI - High iron stores may predict development of type 2 diabetes PMID- 9732365 TI - Danish data do not suggest greatly increased risk of testicular cancer in biopsied testes PMID- 9732366 TI - Training for general practice should be more like that for specialties PMID- 9732368 TI - Inhibition of cholesterol biosynthesis in primary cultured rat hepatocytes by artichoke (Cynara scolymus L.) extracts. AB - High-dose aqueous extracts from artichoke leaves were found to inhibit cholesterol biosynthesis from 14C-acetate in primary cultured rat hepatocytes in a concentration-dependent biphasic manner with moderate inhibition (approximately 20%) between 0.007 and 0.1 mg/ml and more strong inhibition at 1 mg/ml. Cytotoxic effects detected by lactate dehydrogenase leakage and the 3-[4, 5-dimethylthiazol 2-yl]-2,5-dephenyl tetrazolium bromide-assay were restricted to higher concentrations. Replacement of 14C-acetate by 14C-mevalonate largely omitted the inhibiting effect of artichoke extracts indicating an inhibition at the level of hydroxymethylglutaryl-CoA-reductase. However, no direct inhibition of this enzyme could be detected and no other enzymic steps later in the biosynthetic pathway for cholesterol seemed to be affected. Instead, inhibition was found to occur in a time-dependent manner, to last for several hours even after washing out the extracts by fresh medium and to be fully reversible within 20 hr after removal of the extracts. In addition, the stimulation of HMGCoA-reductase activity by insulin was efficiently blocked by the extracts, although other insulin-dependent phenomena, such as increased lactate production, were not influenced. These results suggest an indirect modulation of hydroxymethylglutaryl-CoA-reductase activity as the most likely inhibitory mechanism of the artichoke extracts. Screening of several known constituents of artichoke extracts revealed that cynaroside and particularly its aglycone luteolin were mainly responsible for inhibition, whereas chlorogenic acid was much less effective and caffeic acid, cynarin and other dicaffeoylquinic acids were without significant influence. Indeed, luteolin also efficiently blocked the insulin effect on cholesterol biosynthesis. In conclusion, these results demonstrate that artichoke extracts may inhibit hepatic cholesterol biosynthesis in an indirect but efficient manner and, thus, may contribute via this action to the recently confirmed hypolipidemic influence of this phytopharmacon in man. PMID- 9732367 TI - Role of 5-HT4 receptors in the mouse passive avoidance test. AB - The effects of the administration of different 5-HT4 receptor antagonists (SDZ 205557, GR 125487) and 5-HT4 receptor agonists (BIMU 1, BIMU 8) on memory processes were evaluated in the mouse passive avoidance test. The administration of SDZ 205557 (10 mg kg-1 i.p.) and GR 125487 (10 mg kg-1 i.p.) immediately after termination of the training session produced an amnesic effect. BIMU 1 (20 mg kg 1 i.p.) and BIMU 8 (30 mg kg-1 i.p.), administered 20 min before the training session, prevented the 5-HT4 receptor antagonist-induced amnesia. In the same experimental conditions BIMU 1 (10 mg kg-1 i.p.; 25 microgram/mouse intracerebroventricularly) and BIMU 8 (30 mg kg-1 i.p.; 30 microgram per mouse intracerebroventricularly) prevented scopolamine (1 mg kg-1 i.p.) and dicyclomine (2 mg kg-1 i.p.) amnesia and, at the dose of 10 and 30 mg kg-1 i.p. respectively, prevented amnesia induced by exposure to a hypoxic environment. At the highest effective doses, none of the drugs impaired motor coordination, as revealed by the rota rod test, or modified spontaneous motility and inspection activity, as revealed by the hole board and Animex tests. The 5-HT3 antagonist ondansetron (0.1-1 mg kg-1 i.p.) was unable to prevent scopolamine-, 5-HT4 antagonist- and hypoxia-induced amnesia. These results suggest that the modulation of 5-HT4 receptors plays an important role in the regulation of memory processes. On these bases, the 5-HT4 receptor agonists could be useful in the treatment of cognitive deficits although 5-HT4 receptor antagonists may represent pharmacological tools for investigation of new potential antiamnesic drugs. PMID- 9732369 TI - Pharmacological characterization of human m1 muscarinic acetylcholine receptors with double mutations at the junction of TM VI and the third extracellular domain. AB - A mutant human m5 receptor containing the mutations of Ser465 to Tyr and Thr466 to Pro showed constitutive activity. By replacing the equivalent Ser388 with Tyr and Thr389 with Pro, we created a mutant human m1 (Hm1) receptor with comparable double mutations. The mutant receptor, Hm1(Ser388Tyr, Thr389Pro), was stably expressed in A9 L cells and displayed enhanced responses to classical muscarinic agonists with significantly increased potencies. Choline, a normal component of growth media, showed an efficacy comparable to acetylcholine and carbachol at Hm1(Ser388Tyr, Thr389Pro) receptors. Methylcarbachol, a selective nicotinic agonist, exhibited partial agonist activity at human m1 wild-type receptors and full agonist activity at Hm1(Ser388Tyr, Thr389Pro) receptors. l-Hyoscyamine inhibited the activities of choline and methylcarbachol. Muscarinic antagonists displayed small reductions in binding affinities, although muscarinic agonists showed greatly increased binding affinities for Hm1(Ser388Tyr, Thr389Pro) receptors. All agonists, including choline and methylcarbachol, showed multiple affinity states at Hm1(Ser388Tyr, Thr389Pro) receptors in the absence of GppNHp. The high affinity binding sites for acetylcholine, arecoline and choline were shifted in the presence of GppNHp. These results suggest that Hm1(Ser388Tyr, Thr389Pro) is conformationally favorable for agonist binding and receptor activation. PMID- 9732370 TI - Substance P induction of itch-associated response mediated by cutaneous NK1 tachykinin receptors in mice. AB - Our experiments were conducted to determine whether substance P (SP) would elicit an itch sensation mediated by mast cells in mice. An intradermal injection of SP (10-135 microgram site-1) into the rostral back of the ICR mouse dose-dependently produced scratching of the injected site. The SP- (135 microgram site-1 = 100 nmol site-1) induced scratching was inhibited by capsaicin (repeated administration) and naloxone; features being similar to itch in humans. SP elicited scratching in mast cell-deficient (WBB6F1 W/Wv) mice as well as control (+/+) mice. Pretreatment with compound 48/80 produced similar degrees of inhibition of SP-induced scratching in mast cell-deficient mice as well as control +/+ and ICR mice. Intradermal injections of the NK1 receptor agonist GR73632 produced dose-dependent scratching, while the NK2 agonist GR64349 and the NK3 agonist senktide were without effects. SP-induced scratching was inhibited by the NK1 receptor antagonists spantide and L-668,169, but not by the NK2 antagonist L-659,877. The results suggest that scratching of the mouse induced by an i.d. injection of SP is itch-associated response. The SP action may be mediated at least partly by cutaneous NK1 receptors, and mast cells may not be key factors in SP-induced itching. PMID- 9732371 TI - Evidence against anandamide as the hyperpolarizing factor mediating the nitric oxide-independent coronary vasodilator effect of bradykinin in the rat. AB - The mediator of nitric oxide-(NO) independent vasodilation attributed to endothelium-derived hyperpolarizing factor remains unidentified although there is evidence for a cytochrome P450-derived eicosanoid. Anandamide, the ethanolamide of arachidonic acid and an endogenous ligand for cannabinoid receptors, was proposed as an endothelium-derived hyperpolarizing factor-mediating mesenteric vasodilation to acetylcholine and the hypotensive effect of bradykinin. Using pharmacological interventions that attenuate responses to bradykinin, we examined the possibility of anandamide as a mediator of the NO-independent vasodilator effect of bradykinin in the rat perfused heart by determining responses to anandamide and arachidonic acid. Hearts were treated with indomethacin to exclude prostaglandins and nitroarginine to inhibit NO synthesis and elevate perfusion pressure. The cannabinoid receptor antagonist, SR 141716A (2 microM), reduced dose-dependent vasodilator responses to anandamide (1-10 microgram) but was without effect on responses to AA (1-10 microgram), bradykinin (10-1000 ng) or cromakalim (1-10 microgram). Inhibition of voltage-dependent Ca++ channels with nifedipine (5 nM) attenuated vasodilation to anandamide and arachidonic acid whereas inhibition of Ca++-activated K+ channels with charybdotoxin (10 nM) reduced responses to arachidonic acid but had no effect on vasodilation induced by anandamide. Inhibition of cytochrome P450 with clotrimazole (1 microM) greatly reduced vasodilator responses to bradykinin with less effect on those to anandamide. Finally, the time course of the coronary vasodilator responses to anandamide and bradykinin were dissimilar. These results argue against a role of anandamide in the vasodilator effect of bradykinin in the rat heart. PMID- 9732372 TI - Concentration-effect relationship of l-propranolol and metoprolol in spontaneous hypertensive rats after exercise-induced tachycardia. AB - The concentration-effect relationship of l-propranolol and dl-metoprolol were investigated in spontaneous hypertensive rats using reduction in exercise-induced tachycardia as a pharmacodynamic endpoint. The influence of protein binding on the effect relationship was also assessed. The rats were assigned to treatment or placebo groups, where each group received three randomly selected consecutively increasing steady-state infusions. Different pharmacodynamic effect models were fitted to the data, using nonlinear mixed effect modeling. The data were best described by a combined effect model, with a sum of an ordinary Imax and a linear model. At the lower concentration range, the ordinary Imax model dominated, although at higher concentrations, the effect was linearly related to the antagonist concentration. The Imax were 83 +/- 6 and 103 +/- 6 beats . min-1 and the IC50 were 18.1 +/- 4.3 and 50.6 +/- 15.2 ng/ml for l-propranolol and dl metoprolol, respectively. The slope in the linear model was steeper for l propranolol than for dl-metoprolol, 28.9 +/- 2.8 and 4.48 +/- 0.39 beats . ml . (min . microgram)-1, respectively. Plasma protein binding of l-propranolol was saturable. The unbound IC50 for l-propranolol was 1.14 +/- 0.27 ng/ml. The concentration-effect relationship of l-propranolol was altered at higher plasma concentrations, due to saturable protein binding. The Imax and the linear concentration-effect relationship may be interpreted as a specific beta antagonist effect and a membrane-stabilizing effect, respectively. Using exercise induced tachycardia as a pharmacodynamic endpoint, to study the effect of beta antagonists in spontaneous hypertensive rats, seems to give reliable results and can be a useful model to extrapolate to humans. PMID- 9732373 TI - The role of dopaminergic systems in the perinatal sensitivity to 3, 4 methylenedioxymethamphetamine-induced neurotoxicity in rats. AB - Our study was aimed at analyzing the basis for the apparent lack of perinatal sensitivity to the serotonergic neurotoxin 3, 4-methylenedioxymethamphetamine (MDMA, "ecstasy"). MDMA (20 mg/kg s. c.) repeatedly administered to rat dams during gestation, did not affect [3H]paroxetine-labeled serotonin (5-HT) transporter density and 5-HT content in the offspring. A single dose of MDMA was then given to pups, not exposed prenatally to MDMA, at different postnatal ages (PND14, 21, 28 and 35). Long-term significant reductions in 5-HT levels in all the brain regions examined were only found at PND35. In a different set of experiments, MDMA administered at PND21 alone or in combination with (R)-1-(2, 5 dimethoxy-4-iodophenyl)2-aminopropane (R-DOI, 0.5 mg/kg s.c.), or L-3,4 dihydroxyphenylalanine (L-DOPA, 80 mg/kg s.c.), caused a significant hyperthermia in the pups. However, only L-DOPA followed by MDMA caused a lasting reduction of 5-HT levels and 5-HT transporter density in the hippocampus and in the frontal cortex. In adult animals, no change in 5-HT levels and 5-HT transporter density in different brain regions was either found when MDMA was given to rats previously lesioned with 6-hydroxydopamine, but a significant reduction was again found in the lesioned animals receiving MDMA in combination with L-DOPA. These results appear to indicate that the hyperthermia induced by MDMA is not sufficient to produce lasting neurotoxic effects on the serotonergic system, at least at PND21, and support an important role for dopamine in the mechanism of neurotoxicity of MDMA, suggesting that an already developed dopaminergic system is necessary for the expression of the serotonergic deficits. PMID- 9732374 TI - Transport of L-valine-acyclovir via the oligopeptide transporter in the human intestinal cell line, Caco-2. AB - It has been reported that conjugating acyclovir, a potent antiviral with low oral bioavailability, to L-valine increases its urinary excretion in rats. However, it was also reported that this increase is not found for the D-valine ester, suggesting that a carrier-mediated mechanism is involved in its intestinal absorption. Therefore, mechanisms involved in the transepithelial transport of L valine-acyclovir were investigated using the intestinal cell line, Caco-2, as a model system for the intestinal epithelium. Only the mucosal-to-serosal transport of acyclovir was increased by conjugation with L-valine (approximately 7-fold), suggesting the involvement of a carrier-mediated mechanism. This conclusion was supported by the finding that this increase was saturable. The mucosal-to-serosal transport of L-valine-acyclovir could be inhibited by L-glycylsarcosine, but not by L-valine, suggesting the involvement of the dipeptide carrier. Also it was found that L-valine-acyclovir inhibits the uptake of cephalexin, a substrate for the oligopeptide transporter. Stability of the esters in either the mucosal or serosal bathing solution is more than 90% after completion of the transport study. However, after transport, the receiver solution contained approximately 90% of acyclovir. Based on these findings it was concluded that absorption of the L-valine ester of acyclovir occurs as a result of uptake by the oligopeptide transporter at the apical cell membrane followed by intracellular hydrolysis of the ester and efflux of acyclovir. PMID- 9732375 TI - Calcium-mediated second messengers modulate the expression of behavioral sensitization to cocaine. AB - To assess the influence of calcium channel antagonists on the expression of behavioral sensitization to cocaine, the L-type calcium channel antagonist diltiazem or the N-type calcium channel antagonist omega-conotoxin GVIA was microinjected into the medial nucleus accumbens before a systemic cocaine challenge injection among rats that were previously treated with daily systemic saline or cocaine injections. The results indicated that both of these drugs attenuated the expression of behavioral sensitization to cocaine. Among saline pretreated rats, diltiazem did not influence the behavioral response to an acute injection of cocaine, whereas omega-conotoxin significantly impaired acute cocaine-induced behavioral hyperactivity. A second series of experiments assessed the influence of protein kinases on the expression of behavioral sensitization to cocaine. Inhibitors of calcium/calmodulin-dependent protein kinase II (KN-93, N [2-[[[3-(4'-chlorophenyl)-2-propenyl]methylamino]methyl]phenyl]-N-( 2 hydroxyethyl)-4'-methoxy-benzenesulfonamide phosphate), protein kinase A (H-89, N [2((p-bromocinnamyl)amino)ethyl]-5-isoquinolinesulfonamide) or calcium-dependent protein kinase C (bisindolymaleimide I, 2-[1-(3-dimethylaminopropyl)-1H-indol-3 yl]-3-(1H-indol-3-yl)-maleimi de) were microinjected into the medial nucleus accumbens before a challenge injection of cocaine among rats repeatedly administered either saline or cocaine. None of the kinase inhibitors influenced the behavioral response induced by cocaine in saline-pretreated rats. Among cocaine-sensitized animals, the microinjection of KN-93 or bisindolymaleimide I blocked the expression of behavioral sensitization to cocaine, whereas H-89 had no effect. Taken together, these results indicate that neuronal calcium, acting via calcium-dependent kinases, promotes the expression of behavioral sensitization to cocaine. PMID- 9732376 TI - Multiple actions of methohexital on hippocampal CA1 and cortical neurons of rat brain slices. AB - To explore the mechanism by which methohexital (MTH) activates epileptiform activity in patients with epilepsy, we examined the effects of MTH on hippocampal CA1 and neocortical neurons via extracellular and whole-cell patch-clamp recordings in rat brain slices. Perfusion of slices with 10 to 100 microM MTH caused no significant change in glutamatergic transmission in the hippocampal CA1 region, but enhanced gamma-aminobutyric acid (GABA)A-mediated inhibitory postsynaptic currents and induced spontaneous inhibitory postsynaptic currents in neocortical and hippocampal CA1 neurons. In addition, MTH induced a tonic, bicuculline-sensitive hyperpolarization in association with increases in membrane conductance, suggesting a direct stimulation of GABAA receptors by MTH. Spontaneous epileptiform activity was not observed in the neocortex and hippocampus after exposure of slices to MTH, neither in the standard in vitro condition nor in the presence of 4-aminopyridine, which promotes rhythmic synaptic activities. We suggest that the activation of epileptiform activity in vivo by MTH may result from increased neuronal synchrony via the potentiation of GABAA-mediated synaptic inhibition. PMID- 9732377 TI - Chronic administration of taurine to aged rats improves the electrical and contractile properties of skeletal muscle fibers. AB - A reduction of resting chloride conductance (GCl) and a decrease of the voltage threshold for contraction are observed during aging in rat skeletal muscle. The above alterations are also observed in muscle of adult rat after taurine depletion. As lower levels of taurine were found by others in aged rats compared to young rats, we tested the hypothesis that a depletion of taurine may contribute to the alteration of the electrical and contractile properties we found in skeletal muscle during aging. This was accomplished by evaluating the potential benefit of a pharmacological treatment with the amino acid. To this aim 25-mo-old Wistar rats were chronically treated (2-3 mo) with taurine (1 g/kg p.o. daily) and the effects of such a treatment were evaluated in vitro on the passive and active membrane electrical properties of extensor digitorum longus muscle fibers by means of current-clamp intracellular microelectrode technique. Excitation-contraction coupling was also evaluated by measuring the voltage threshold for contraction with the intracellular microelectrode "point" voltage clamp method. In parallel muscle and blood taurine contents were determined by high-performance liquid chromatography. Taurine supplementation significantly raised taurine content in muscle toward that found in adult rats. Supplementation also significantly increased GCl vs. the adult value, in parallel the excitability characteristics (threshold current and latency) related to this parameter were ameliorated. The increase of GCl induced by taurine was accompanied by a restoration of the pharmacological sensitivity to the R(+) enantiomer of 2-(p-chlorophenoxy) propionic acid, a specific chloride channel ligand. In parallel also the protein kinase C-mediated modulation of the channel was restored; in fact the potency of 4-beta-phorbol 12, 13-dibutyrate in reducing GCl was lower in taurine-treated muscles vs. untreated aged, being rather similar to that observed in adult. The treatment also improved the mechanical threshold for contraction of striated fibers which in aged rats is shifted toward more negative potentials, moving it toward the adult values. Our results suggest that the reduction of taurine content could play a role in the alteration of electrical and contractile properties observed during aging. These findings may indicate a potential application of taurine in ensuring normal muscle function in the elderly. PMID- 9732378 TI - CVT-124, a novel adenosine A1 receptor antagonist with unique diuretic activity. AB - Administration of the selective adenosine A1 receptor antagonist, CVT-124, to conscious chronically instrumented rats resulted in significant increases in urine flow rate and sodium excretion without affecting potassium excretion or renal hemodynamics. Its maximum effect was twice that of hydrochlorothiazide which was associated with a significant kaliuresis. The diuretic effect of CVT 124 was less than that observed with furosemide; however, furosemide administration was associated with a large increase in potassium excretion as well as a reduction in glomerular filtration rate. When given at equinatriuretic doses, CVT-124 enhanced the diuretic and natriuretic activity of furosemide without further increasing potassium excretion. In contrast, the combination of hydrochlorothiazide and furosemide resulted in a 3-fold increase in potassium excretion. These data suggest that CVT-124 possesses unique diuretic activity and, as such, it represents a potential new therapeutic in fluid retaining disorders. In addition, its unique mechanism of action suggests that CVT-124 would be effective in otherwise diuretic-resistant patients. PMID- 9732379 TI - Comparing the subjective, psychomotor and physiological effects of intravenous pentazocine and morphine in normal volunteers. AB - The purposes of this study were to characterize the subjective, psychomotor and physiological effects of pentazocine in non-drug-abusing volunteers and to compare and contrast the effects of pentazocine with those of morphine. Sixteen subjects without histories of opiate dependence were injected in an upper extremity vein with 0, 7.5, 15 or 30 mg/70 kg pentazocine or 10 mg/70 kg morphine, using a randomized, double-blind, crossover design. Pentazocine increased scores on the pentobarbital-chlorpromazine-alcohol group and lysergic acid diethylamide scales and decreased scores on the benzedrine group scale of the Addiction Research Center Inventory, increased adjective checklist ratings of "nodding," "sweating" and "turning of stomach" and increased visual analog scale ratings of "difficulty concentrating," "drunk" and "having unpleasant bodily sensations." Pentazocine (30 mg) had a greater propensity to increase ratings associated with dysphoria than did 10 mg of morphine. Pentazocine produced impairment on four measures of psychomotor performance. Ten milligrams of morphine produced minimal psychomotor impairment. Both pentazocine and morphine induced miosis, but 10 mg of morphine had a greater magnitude of effect than 30 mg of pentazocine. The results of the present study demonstrate that 7.5 to 30 mg of pentazocine had orderly, dose-related effects on subjective, psychomotor and physiological variables. Further, a clinically relevant dose of pentazocine, 30 mg, produced a greater magnitude of dysphoric subjective effects than did 10 mg of morphine, which is consistent with the literature reporting that pentazocine has a greater likelihood of inducing psychotomimesis than do other opioids. PMID- 9732380 TI - Hydrogen peroxide-induced stimulation of L-type calcium current in guinea pig ventricular myocytes and its inhibition by adenosine A1 receptor activation. AB - Hydrogen peroxide (H2O2) produces complex cardiac effects that may involve altered calcium homeostasis. The cardiotoxic effects of H2O2 can be attenuated by adenosine A1 receptor agonists. The present study examined the effect of H2O2 on L-type Ca++ current (ICa,L) in guinea pig ventricular myocytes under two different recording conditions and the influence of adenosine receptor agonists. H2O2 (100 microM), did not have any significant effect on ICa,L, under conventional whole cell patch configuration. However, when recorded under nystatin perforated patch configuration, H2O2 caused a gradual and significant increase (84 +/- 14%) in ICa,L compared to control values. N6 cyclopentyladenosine (CPA), an adenosine A1 receptor agonist, significantly attenuated the effect of H2O2. The inhibitory effect of N6-cyclopentyladenosine was antagonized by 8cyclopentyl-1, 3-dipropylxanthine, an adenosine A1 receptor antagonist. The A2A and A3 receptor agonists, 2-p-(2-Carboxyethyl)phenethylamino 5'- N - ethylcarboxamidoadenosine (CGS-21680) and 1-deoxy-1-[6-[[(3 iodophenyl)methyl]amino]-9H-purin-9-yl]-N-methyl-be ta-D-ribofuranuronamide, respectively, did not modulate the enhancement of ICa,L by H2O2. Moreover the effects of N6-cyclopentyladenosine were mimicked by the protein kinase C inhibitor bisindolylmaleimide. Thus, our results demonstrate a potent stimulatory effect of H2O2 on ICa,L in guinea pig ventricular myocytes. We further demonstrate that adenosine A1 receptor activation attenuates this effect. Our results suggest a potential basis for altered calcium homeostasis in response to H2O2 as well as the salutary effects of A1 receptor activation against H2O2 induced cardiotoxicity. PMID- 9732381 TI - Effect of KATP channel blocker U37883A on renal function in experimental diabetes mellitus in rats. AB - An increase in glomerular filtration rate (GFR) in early diabetes mellitus is considered a risk factor for the development of diabetic nephropathy. Insulin deficiency may increase the activity of ATP-sensitive potassium channels (KATP), which could promote afferent arteriolar vasodilation und thus contribute to glomerular hyperfiltration in early diabetes mellitus. To further elucidate this hypothesis we performed renal clearance experiments in anesthetized rats at 2 and 6 weeks after onset of streptozotocin-induced insulin-treated diabetes mellitus and studied the acute effect of the putative KATP channel blocker 4 morpholinecarboximidine-N-1-adamantyl-N'-cyclohexylhydr ochloride (U37883A) on renal function. In control rats, application of U37883A (1.5 mg/kg i.v. bolus plus 1.5 mg/kg/hr) induced a significant reduction in heart rate, but did not affect or even slightly increased mean arterial blood pressure. Furthermore, U37883A did not significantly affect renal vascular resistance, renal blood flow or GFR, but caused an eukaliuretic diuresis and natriuresis and lowered plasma renin activity. Diabetic rats at both 2 or 6 weeks after streptozotocin exhibited essentially an identical response to U37883A; in particular, RVR and glomerular hyperfiltration remained unchanged. These results show that in both control and diabetic rats, the renal excretory function, renin secretion and pace setting in the heart were sensitiv to U37883A, implying a functional contribution of KATP channel activity. However, in both control and diabetic rats, renal vascular resistance, renal blood flow, or GFR were not altered by U37883A. These results argue against a substantial role for KATP channels in the basal control of renal hemodynamics in both nondiabetic and diabetic rats. PMID- 9732382 TI - Inhibitory effects of nitric oxide donors on nitric oxide synthesis in rat gastric myenteric plexus. AB - We investigated whether nitric oxide (NO) exerts an inhibition on its own synthesis in the gastric myenteric plexus in rats. Nonadrenergic, noncholinergic relaxations in response to transmural electrical stimulation (TS) were markedly antagonized by NG-nitro-L-arginine methyl ester, (10(-4) M) and abolished by tetrodotoxin (10(-6) M). Pretreatment with various NO donors (3-morpholino sydnonymide [SIN-1 (3 x 10(-7) to 3 x 10(-6) M)], S-nitroso-N-acetylpenicillamine (10(-6) to 10(-5) M), sodium nitroprusside (10(-8) to 3 x 10(-8) M) and 8 bromoquanosine 3', 5'-cyclic monophosphate [8-bromo-cGMP (10(-6) to 3 x 10(-6) M)]) significantly inhibited TS-evoked nonadrenergic, noncholinergic relaxations in a dose-dependent manner. In contrast, vasoactive intestinal polypeptide (10( 8) M)-induced relaxations were not affected by SIN-1 or 8-bromo-cGMP. TS evoked a significant increase in 3H-citrulline formation, which was completely abolished by calcium-free medium, NG-nitro-L-arginine methyl ester, (10(-4) M) and tetrodotoxin (10(-6) M). 3H-citrulline formation evoked by TS was significantly inhibited by SIN-1 (10(-7) to 10(-5) M) and 8-bromo-cGMP (10(-7) to 10(-5) M) in a dose-dependent manner. The inhibitory effect of SIN-1 was partially prevented by 1H-[1,2, 4]oxadiazolo[3,4-a]quinoxalin-1-one (10(-5) M), a guanylate cyclase inhibitor. We conclude that NO synthesis in the gastric myenteric plexus is negatively regulated by NO and cGMP. This suggests an autoregulatory feedback mechanism of NO synthesis in the gastric myenteric plexus. PMID- 9732383 TI - Characterization of the effects of the partial dopamine agonist terguride on cocaine self-administration in the rat. AB - Dopamine neurotransmission is an important neuropharmacological component of cocaine self-administration in rodents. Terguride is a prototype drug belonging to a recently characterized class of compounds, dopamine partial agonists, which appear to possess a unique pharmacological profile in altering dopamine neurotransmission, where these drugs act as antagonists in conditions of high dopaminergic tone. The aim of the present study was therefore to test the effects of systemic administration of terguride in rats self-administering cocaine intravenously. The different aspects of cocaine self-administration examined after treatment with terguride were (a) the acute reinforcing properties of cocaine in rats exposed to limited-access self-administration of cocaine, (b) a full cocaine dose-effect function, (c) the reinforcing properties of cocaine as measured by a progressive ratio schedule and (d) the ability of terguride to maintain self-administration by itself. Terguride (0.025-0.4 mg/kg i.p.) significantly and dose-dependently reduced the acute reinforcing properties of cocaine as measured by an increase in responding for a single training dose of cocaine and a reduction of the inter-reinforcement interval. In addition, terguride (0.2-0.4 mg/kg) shifted the entire cocaine dose-effect function to the right, thus showing an antagonism of the reinforcing properties of cocaine independent of response rate. Moreover, in rats trained to self-administer cocaine on a progressive ratio schedule, terguride reduced the maximum fixed ratio ("breaking point") for cocaine reinforcement, also suggesting a decrease in the reinforcing properties of cocaine. Finally, in rats trained to self administer cocaine terguride did not substitute for cocaine, thus indicating that terguride does not maintain intravenous self-administration by itself. PMID- 9732385 TI - Dopamine D2 receptors mediate glomerular hyperfiltration due to amino acids. AB - Renal dopamine has been proposed to be involved in the regulation of glomerular filtration rate (GFR). Because inhibition of dopamine D2 receptors abolishes the renal hyperfiltration due to amino acid load, we tested the hypothesis that pharmacological activation of D2-like receptors mimicked this renal response. In anesthetized rats, quinpirole (0.3 microgram . 100 g-1 . min-1), an agonist for receptors of the D2-like family, caused an increase in GFR by 20 +/- 2%, which corresponded to that provoked by infusion of an 10% amino acid solution. The D2 receptor antagonist S(-)-sulpiride that acts both centrally and peripherally completely abolished the renal hemodynamic response to quinpirole and to amino acids whereas domperidone, a peripherally acting D2 receptor antagonist, inhibited this hyperfiltration only in part. Urinary dopamine excretion increased in response to amino acid infusion whether GFR increased or not. We conclude that, in anesthetized rats, dopamine D2 receptors contribute to the amino acid induced hyperfiltration and that both central and peripheral receptors might be involved, whereas dopamine excreted into the urine does not appear to play a functional role in this renal hemodynamic response. PMID- 9732384 TI - Full agonistic properties of BAY x 3702 on presynaptic and postsynaptic 5-HT1A receptors electrophysiological studies in the rat hippocampus and dorsal raphe. AB - The present studies evaluated the effects of acute and long-term administration of the 5-HT1A agonist BAY x 3702 on the responsiveness of dorsal raphe 5-HT neurons and of dorsal hippocampus CA3 pyramidal neurons. BAY x 3702 potently reduced the firing activity of 5-HT neurons and of CA3 pyramidal neurons when applied by microiontophoresis and this inhibitory effect of BAY x 3702 was fully antagonized by low intravenous doses of the 5-HT1A antagonist WAY 100635. Concurrent microiontophoretic application of BAY x 3702 did not antagonize the suppressant effect of 5-HT on firing activity of 5-HT and CA3 pyramidal neurons. Sustained administration of BAY x 3702 for 2 days (1 and 1.25 mg/kg/day using osmotic minipumps implanted subcutaneously) markedly decreased the firing rate of dorsal raphe 5-HT neurons. This was followed by a full recovery to normal after only 7 days of treatment. The postsynaptic 5-HT1A receptors in the hippocampus, contrary to the presynaptic 5-HT1A receptors, were not desensitized after a 14 day treatment. In conclusion, BAY x 3702 acted as a full and potent agonist both at somatodendritic 5-HT1A autoreceptors and at postsynaptic 5-HT1A receptors. Long-term administration of BAY x 3702 resulted in a desensitization of the somatodendritic 5-HT1A autoreceptors, but in an unaltered responsiveness of 5 HT1A receptors on pyramidal neurons. These results suggest that sustained administration of BAY x 3702 enhances neurotransmission at postsynaptic 5-HT1A receptors. PMID- 9732386 TI - Further characterization of the expression in liver and catalytic activity of CYP2B6. AB - Previous studies in this laboratory have determined the lack of specificity of several antibody and substrate probes of CYP2B6. The goals of the current study were to examine the expression of CYP2B6 in a bank of human liver microsome (HLM) samples using a new specific monoclonal antibody (MAb 49-10-20) and to further characterize the substrate specificity of CYP2B6. A 100-fold variability in expression of immunodetectable CYP2B6 was demonstrated in a bank of 19 HLM samples (0.7 pmol/mg protein to 71. 1 pmol/mg protein) using MAb 49-10-20. CYP2B6 levels were found to significantly (P < .0001) correlate with S-mephenytoin N demethylation to nirvanol (r2 = 0.89), 7-hydroxy-4-trifluoromethylcoumarin formation (r2 = 0.81) and several markers of CYP3A levels and activity. The relationships between nirvanol formation and CYP3A levels or activity were found to depend on two HLM samples. Km (apparent) values were generated for benzyloxyresorufin O-deethylation (1.3 microM), benzphetamine N-demethylation (93.4 microM), 3-cyano 7-ethoxycoumarin O-deethylation (71.3 microM), midazolam 1'-hydroxylation (46.1 microM) and 4-chloromethyl-7-ethoxycoumarin O-deethylation (33.7 microM) using expressed CYP2B6. Testosterone 16beta-hydroxylation by expressed CYP2B6 resulted in atypical kinetics characteristic of substrate activation. The data best fit the Hill equation with a Km (apparent) of 50.5 microM and an n of 1.3 (n = number of sites bound by activator). In conclusion, the highly specific MAb 49-10-20 was used to provide further confirmation that S mephenytoin N-demethylation to nirvanol is a CYP2B6 selective probe. Finally, some, but not all substrates of CYP2B6 demonstrate autoactivation. PMID- 9732387 TI - Effects of N-methyl-D-aspartate receptor antagonists on discriminative stimulus effects of naloxone in morphine-dependent rats using the Y-maze drug discrimination paradigm. AB - The present study assessed the ability of various site-selective N-methyl-D aspartate (NMDA) receptor antagonists to affect the discriminative stimulus properties of naloxone in morphine-dependent rats. Adult male Wistar rats were trained to discriminate 0.1 mg/kg of s.c. naloxone from saline using a Y-maze shock-avoidance procedure. Naloxone-appropriate responding was exhibited as a function of naloxone dose (0.01-1.0 mg/kg, ED50 = 0.03 mg/kg) and was also observed when morphine treatment temporarily was discontinued (8-96 hr, peak at 24 hr). Discriminative stimulus effects of naloxone (0.1-3.0 mg/kg) were antagonized by morphine (10-100 mg/kg). Ligands of peripheral opioid receptors failed to either substitute for naloxone (methylnaloxone, 0.1-3.0 mg/kg) or attenuate naloxone's stimulus effects (loperamide, 1-30 mg/kg). In rats treated with the training dose of naloxone, administration of dizocilpine (0.03-0.3 mg/kg) and D-CPPene (1-10 mg/kg) decreased levels of naloxone-appropriate responding, whereas memantine (1-30 mg/kg), ACEA-1021 (10 and 50 mg/kg) and eliprodil (3-30 mg/kg) seemed to have little or no effects. Meanwhile, all NMDA receptor antagonists produced a decrease in the occurrence of two or more of the following opioid withdrawal signs: weight loss, forelimb tremor, ptosis, diarrhea and "wet-dog"-like shaking. Additionally, dizocilpine (0.1 mg/kg), D-CPPene (5.6 mg/kg) and ACEA-1021 (50 mg/kg) but not memantine (10 mg/kg) or eliprodil (30 mg/kg) significantly reduced the naloxone-appropriate escape area selection when administered during the period of suspended morphine treatment 24 hr after the last morphine injection. Thus, NMDA receptor antagonists appear to inhibit the discriminative stimulus effects of both naloxone-precipitated and spontaneous morphine withdrawal, and this ability depends on the type of antagonist applied. PMID- 9732388 TI - Potentiation and inhibition of nicotinic acetylcholine receptors by spermine in the TE671 human muscle cell line. AB - Nicotinic acetylcholine receptors (nAChR) of the TE671 cell line were investigated using whole-cell and membrane patch recording techniques. At negative holding potentials (VH), pulses of acetylcholine (ACh) elicited whole cell inward currents that rapidly desensitized. The EC50 value for ACh at VH = 60 mV was 7.8 microM. The ACh-induced current reversed at approximately 0 mV. Desensitization of nAChR by ACh was biphasic and reversible within approximately 20 sec. Spermine (1-100 microM) potentiated responses to ACh (10 microM - 1 mM) by reducing the rate of onset of desensitization; potentiation was inhibited by arcaine (10-100 microM). Spermine (1 mM) noncompetitively antagonized the AChinduced current. Antagonism by 1 to 5 mM spermine was voltage-dependent, increasing with negative VH. In 100 microM arcaine, this antagonism was shown to contain a voltage-independent component. Spermine (10 mM) increased the EC50 values for ACh, suggesting that at this concentration the polyamine is also a competitive antagonist. Single channel openings elicited during application of ACh to outside-out patches had a conductance of 47 pS at VH = -60 mV. At 10 and 100 microM, spermine increased channel open probability (po), but at 1 mM spermine, po was not significantly different from controls. The single channel conductance for ACh was unaffected by 10 and 100 microM spermine, but was decreased by 1 mM spermine. Spermine promoted the occurrence of approximately 27 pS openings. It is proposed that spermine acts at an excitatory modulatory site similar to that present on N-methyl-D-aspartate receptors and at least three inhibitory sites on nAChR of TE671 cells. PMID- 9732389 TI - Antiplatelet efficacy of XV459, a novel nonpeptide platelet GPIIb/IIIa antagonist: comparative platelet binding profiles with c7E3. AB - Recent advances in the development of i.v. platelet glycoprotein alphaIIb/beta3 integrin (GPIIb/IIIa) antagonists led to the development of either a class of small-molecular-weight antagonists with a short to ultra-short duration of antiplatelet effects (Integrelin, Tirofiban, DMP728) or a very long-acting antagonist (ReoPro). Thus the present study was undertaken to characterize the antiplatelet efficacy of a small-molecule GPIIb/IIIa antagonist, DMP754/XV459, and to determine its platelet GPIIb/IIIa receptor binding profiles. DMP754, upon its conversion with esterases to its free acid form XV459, and XV459 itself, demonstrated high potency (IC50 = 0.030-0.060 microM) in inhibiting human platelet aggregation induced by ADP (100 microM), thrombin receptor agonist peptide (10 microM) or collagen (20 microgram/ml) in citrate or heparin. Maximal platelet aggregation inhibition was achieved at 50 to >/=80% receptor occupancy, depending on the agonist used. Both XV459 and c7E3 bind with high affinity to either activated human platelets (Kd = 0.0008 and 0.0091 microM, respectively) or unactivated human platelets (Kd = 0.0025 and 0.0092 microM, respectively). XV459 demonstrated tight association with human, baboon and (to a lesser extent) canine platelets (t1/2 of dissociation = 7 +/- 0, 8 +/- 1 and 1.4 +/- 0.1 minutes, respectively). Both c7E3 and XV459 associate tightly with slower dissociation rates to unactivated human platelets. XV459 represents a potent antiplatelet agent in inhibiting platelet aggregation along with offering high affinity and a relatively slow dissociation rate from human platelet GPIIb/IIIa receptors that might allow for once-a-day p.o. dosage. PMID- 9732390 TI - Application of liquid chromatography/mass spectrometry in accelerating the identification of human liver cytochrome P450 isoforms involved in the metabolism of iloperidone. AB - Iloperidone, [1-[4-[3-[4-(6-fluoro-1, 2-benzisoxazol-3-yl)-1-piperidinyl]propoxy] 3-methoxyphenyl]eth anone, 1, is currently undergoing clinical trials as a potential antipsychotic agent. The metabolism of iloperidone was studied in human liver microsomes to define the metabolic pathways and to identify the cytochrome P450 (CYP) isoforms responsible for the formation of major iloperidone metabolites. Iloperidone was extensively metabolized in vitro via hydroxylation, reduction and O-demethylation to produce 1-[4-[3-[4-(6-fluoro-1, 2-benzisoxazol-3 yl)-1-piperidinyl]propoxy]-3-methoxyphenyl]-2- hydrox yethanone, 4; 4-[3-[4-(6 fluoro-1, 2-benzisoxazol-3-yl)-1-piperidinyl]propoxy]-3-methoxy-alpha-met hylben zene methanol, 3, and 1-[4-[3-[4-(6-fluoro-1, 2-benzisoxazol3-yl)-1 piperidinyl]propoxy]-3-hydroxyphenyl]etha none, 2, respectively, in decreasing order of abundance. The major in vitro metabolite, 4, present in trace quantities in urine, was postulated to be either eliminated in bile as a conjugate or further metabolized to a phenol, 4-[3-[4-(6-fluoro-1, 2-benzoisoxazol-3-yl) piperidin1-yl]propoxy]-3-methoxyphenol , 5. The formation of the three major in vitro metabolites 2, 3 and 4 was NADPH dependent. The major circulating and urinary metabolite in humans dosed with 1 was metabolite 3. The mean apparent Km and Vmax for formation of 2 by human liver microsomes was 7.4 +/- 3.0 microM and 0.0343 +/- 0.0134 nmol min-1 mg-1, respectively. The mean apparent Km and Vmax for 3 was 101.2 +/- 34.7 microM and 0.1414 +/- 0.0346 nmol min-1 mg-1, respectively. The mean apparent Km and Vmax for 4 was 39.7 +/- 10.8 microM and 0.1372 +/- 0.056 nmol min-1 mg-1, respectively. The CYP isoenzymes responsible for the formation of metabolites 2, 3 and 4 were determined by using selective chemical inhibitors and by correlation studies. Metabolites 2 and 4 were formed by CYP3A4 and by the polymorphic CYP2D6 respectively. Metabolite 3 is postulated to be produced mainly by a cytosolic enzyme(s), although CYP3A, CYP1A2 and CYP2E1 isozymes were shown to be involved in its formation as well. The power of liquid chromatography/mass spectrometry in greatly accelerating the process of identifying the human liver CYP isoforms involved in the metabolism of iloperidone was demonstrated in this study. Liquid chromatography/mass spectrometry was used in the initial studies to confirm the identities of the metabolites. This was followed by accurate and reliable quantitation of individual metabolites present in biological extracts by operating the mass spectrometer in the selected ion monitoring mode. PMID- 9732391 TI - A study on the metabolism of etoposide and possible interactions with antitumor or supporting agents by human liver microsomes. AB - The metabolism of etoposide was investigated by using human liver microsomes and nine recombinant human cytochrome P450 (CYP) isoforms to identify the CYP isoform(s) involved in the major metabolic pathway (3'-demethylation) of etoposide as well as to evaluate the possible metabolic interactions with several antitumor or supporting agents. The 3'-demethylation of etoposide followed a Michaelis-Menten one-enzyme kinetic behavior in six human liver microsomal samples. The relationships were assessed with six different human liver microsomes between the 3'-demethylation of etoposide and metabolic activities for substrate probes of the respective CYP isoforms, showing a significant correlation (r = 0. 932, P < .01) only with 6beta-hydroxylation of testosterone, a marker substrate for CYP3A4. Inhibitor/substrate probes for CYP3A4, ketoconazole, troleandomycin, verapamil and cyclosporin, or supporting agents, vincristine and prednisolone, inhibited etoposide 3'-demethylation by human liver microsomes. p-Nitrophenol, a substrate for CYP2E1, also inhibited etoposide 3' demethylation. Among the nine recombinant human CYP isoforms, CYP3A4 exhibited the highest catalytic activity with respect to etoposide 3'-demethylation, compared with the minor activities of CYP1A2 and 2E1. Collectively, these data suggest that etoposide 3'-demethylation is mediated mainly by CYP3A4 and to a minor extent by CYP1A2 and 2E1. Furthermore, some supporting agents (vincristine and prednisolone) and the substrates of CYP3A4, which may be coadministered with etoposide during the cancer chemotherapies, inhibit the etoposide 3' demethylation activity in vitro. The results may provide clinical implications with respect to the possible metabolic interactions between etoposide and other drugs studied herein in patients with cancer undergoing etoposide concurrently with either of them. PMID- 9732392 TI - Differential blockade of the antinociceptive effects of centrally administered cannabinoids by SR141716A. AB - We evaluated delta-9 tetrahydrocannabinol (Delta9-THC), delta-8 tetrahydrocannabinol (Delta8-THC), CP55,940 (CP55), 1-deoxy-11-hydroxy-Delta8-THC dimethylheptyl (deoxy-HU210, a CB2-selective cannabinoid that also binds the CB1 receptor) and the endogenous cannabinoid anandamide (ANA) via i.c.v. and/or intrathecal (i.t.) routes of administration, alone and in combination with SR141716A (SR), a CB1 antagonist, using the tail-flick test. Our studies were performed in order better to characterize potential diversity in interactions of the cannabinoids with the cannabinoid (CB1) receptor. When SR was administered i.c.v. or i.p. before Delta9-THC, Delta8-THC or CP55 (i.c.v. or i.t.), SR was a potent antagonist and the blockade was complete (AD50 50% MPE) were achieved using the tail flick test whereas base line levels of antinociception were observed 30 sec later in the same rats using the hotplate test. By contrast, antinociception evoked by i.v. morphine (10 micromol) exceeded 50% MPE using both the hotplate and tail flick tests although the "apparent" potency was approximately 2.5 times greater using the tail flick test. After i.c.v. dosing, M6G (0.22-3.3 nmol) was significantly (P < .05) more potent when assessed using the tail flick compared with the hotplate test. Taken together, these data strongly indicate that following central and systemic administration, M6G's antinociceptive effects are mediated primarily by spinal structures whereas both spinal and supraspinal mechanisms contribute to systemic morphine's antinociceptive effects. PMID- 9732397 TI - Interaction of 2',2'-difluorodeoxycytidine (gemcitabine) and formycin B with the Na+-dependent and -independent nucleoside transporters of Ehrlich ascites tumor cells. AB - The uptake of [3H]formycin B by Ehrlich ascites tumor cells was examined in both normal Na+ buffer (physiological) and nominally Na+-free buffer (iso-osmotic replacement with Li+). These studies were conducted to further characterize the equilibrative nucleoside transporter subtypes of Ehrlich cells and to assess the contribution of Na+-dependent concentrative transport mechanisms to the cellular accumulation of nucleoside analogues by these cells. Formycin B is poorly metabolized by mammalian cells and, hence, can be used as a substrate to measure transport kinetics in energetically competent cells. Initial studies established that formycin B inhibited [3H]uridine uptake by the ei (equilibrative inhibitor insensitive) and es (equilibrative inhibitor-sensitive) transporters of Ehrlich cells with Ki values of 48 +/- 28 and 277 +/- 25 microM, respectively. Similarly, [3H]formycin B had Km values of 111 +/- 52 and 635 +/- 147 microM for uptake by the ei and es transporters, respectively. When assays were conducted in the presence of Na+, plus 100 nM nitrobenzylthioinosine to prevent efflux via the es transporters, the intracellular concentration of [3H]formycin B exceeded the initial medium concentration by more than 3-fold, indicating the activity of a Na+-dependent transporter. Interestingly, the initial rate of uptake of [3H]formycin B was significantly higher in the Li+ buffer (es-mediated Vmax = 65 +/- 10 pmol/microliter . sec) than in the Na+ buffer (Vmax = 8.4 +/- 0.9 pmol/microliter . sec); this may reflect trans-acceleration of [3H]formycin B uptake by elevated intracellular adenosine levels resulting from the low Na+ environment. This model was then used to assess the interaction of gemcitabine (2',2'-difluorodeoxycytidine) with the equilibrative and concentrative nucleoside transporters. Gemcitabine, which has shown considerable potential for the treatment of solid tumors, was a relatively poor inhibitor of [3H]formycin B uptake via the equilibrative transporters (IC50 approximately 400 microM). In contrast, gemcitabine was a potent inhibitor of the Na+-dependent nucleoside transporter of Ehrlich cells (IC50 = 17 +/- 5 nM). These results suggest that the cellular expression/activity of Na+-dependent nucleoside transporters may be an important determinant in gemcitabine cytotoxicity and clinical efficacy. PMID- 9732398 TI - S 16924 ((R)-2-[1-[2-(2,3-dihydro-benzo[1,4] dioxin-5-Yloxy)-ethyl]-pyrrolidin 3yl]-1-(4-fluoro-phenyl)-ethanone), a novel, potential antipsychotic with marked serotonin (5-HT)1A agonist properties: I. Receptorial and neurochemical profile in comparison with clozapine and haloperidol. AB - S 16924 showed a pattern of interaction at multiple (>20) native, rodent and cloned, human (h) monoaminergic receptors similar to that of clozapine and different to that of haloperidol. Notably, like clozapine, the affinity of S 16924 for hD2 and hD3 receptors was modest, and it showed 5-fold higher affinity for hD4 receptors. At each of these sites, using a [35S]GTPgammaS binding procedure, S 16924, clozapine and haloperidol behaved as antagonists. In distinction to haloperidol, S 16924 shared the marked affinity of clozapine for h5-HT2A and h5-HT2C receptors. However, an important difference to clozapine (and haloperidol) was the high affinity of S 16924 for h5-HT1A receptors. At these sites, using a [35S]GTPgammaS binding model, both S 16924 and clozapine behaved as partial agonists, whereas haloperidol was inactive. In vivo, the agonist properties of S 16924 at 5-HT1A autoreceptors were revealed by its ability to potently inhibit the firing of raphe-localized serotoninergic neurones, an action reversed by the selective 5-HT1A receptor antagonist, WAY 100,635. In contrast, clozapine and haloperidol only weakly inhibited raphe firing, and their actions were resistant to WAY 100,635. Similarly, S 16924 more potently inhibited striatal turnover of 5-HT than either clozapine or haloperidol. Reflecting its modest affinity for D2 (and D3) autoreceptors, S 16924 only weakly blocked the inhibitory influence of the dopaminergic agonist, apomorphine, upon the firing rate of ventrotegmental area-localized dopaminergic neurones. Further, S 16924 only weakly increased striatal, mesolimbic and mesocortical turnover of dopamine (DA). Clozapine was, similarly, weakly active in these models, whereas haloperidol, in line with its higher affinity at D2 (and D3) receptors, was potently active. In the frontal cortex (FCX) of freely moving rats, S 16924 dose dependently reduced dialysate levels of 5-HT, whereas those of DA and NAD were dose-dependently increased in the same samples. In contrast, although S 16924 also suppressed 5-HT levels in the striatum and nucleus accumbens, DA levels therein were unaffected. Clozapine mimicked this selective increase in DA levels in the FCX as compared to striatum and accumbens. In contrast, haloperidol modestly increased DA levels in the FCX, striatum and accumbens to the same extent. In distinction to S 16924, clozapine and haloperidol exerted little influence upon 5-HT levels. Finally, the influence of S 16924 upon FCX levels of 5-HT, DA (and NAD) was attenuated by WAY 100,635. In conclusion, S 16924 possesses a profile of interaction at multiple monoaminergic receptors comparable to that of clozapine and distinct to that of haloperidol. In addition, S 16924 is a potent, partial agonist at 5-HT1A receptors. Correspondingly, acute administration of S 16924 decreases cerebral serotoninergic transmission and selectively reinforces frontocortical as compared to subcortical dopaminergic transmission. In line with these actions, S 16924 shows a distinctive profile of activity in functional (behavioral) models of potential antipsychotic activity (companion paper). PMID- 9732399 TI - S 16924 ((R)-2-[1-[2-(2,3-dihydro-benzo[1,4] dioxin-5-yloxy)-ethyl]-pyrrolidin 3yl]-1-(4-fluoro-phenyl)-ethanone), a novel, potential antipsychotic with marked serotonin (5-HT)1A agonist properties: II. Functional profile in comparison to clozapine and haloperidol. AB - S 16924 antagonized locomotion provoked by dizocilpine and cocaine, reduced conditioned avoidance responses and blocked climbing elicited by apomorphine, models predictive of control of the positive symptoms of schizophrenia: its median inhibitory dose (ID)50 was 0.96 mg/kg, s.c. vs. 1.91 for clozapine and 0.05 for haloperidol. Rotation elicited in unilateral, substantia nigra-lesioned rats by the D1 agonist, SKF 38393, and by the D2 agonist, quinpirole, was blocked equipotently by S 16924 (0.8 and 1. 7) and clozapine (0.6 and 2.0), whereas haloperidol preferentially blocked quinpirole (0.02) vs. SKF 38393 (1.8). S 16924 more potently inhibited the head-twitches elicited by 1-(2, 5-dimethoxy-4 iodophenyl)-2-aminopropane (DOI) and the locomotion provoked by phencyclidine than it inhibited the locomotion elicited by amphetamine (ID50s = 0.15 and 0.02 vs. 2.4). Clozapine showed a similar preference (0.04 and 0.07 vs. 8.6), but not haloperidol (0. 07 and 0.08 vs. 0.04). The discriminative stimulus (DS) properties of DOI were also blocked by S 16924 (ID50 = 0.17) and clozapine (0. 05) but not by haloperidol (>0.16). S 16924 fully (100%) generalized [effective dose (ED)50 = 0.7] to a clozapine DS and clozapine (0.23) fully generalized to a S 16924 DS whereas haloperidol (>/=0.08) only partially generalized (/=80.0) or clozapine (>/=80.0). Further, S 16924 (ID50 = 3.2) and clozapine (5.5) inhibited induction of catalepsy by haloperidol. This action of S 16924 was abolished by the 5-HT1A receptor antagonist, WAY 100,635 (0.16), which less markedly attenuated the anticataleptic action of clozapine. Further, although gnawing elicited by methylphenidate was inhibited by S 16924 (ID50 = 8.4), clozapine (19.6) and haloperidol (0.04), only the action of S 16924 was blocked by WAY 100,635 (0.16). Haloperidol potently (0.01-0.16, approximately 24-fold) increased prolactin levels whereas they were less markedly affected by S 16924 (2.5-40.0, 4-fold) and clozapine (10.0-40.0, 3-fold). Clozapine displayed high affinity at cloned, human, muscarinic (M1) and native, histamine (H1) receptors (Kis = 4.6 and 5.4 nM, respectively), whereas S 16924 (>1000 and 158) and haloperidol (>1000 and 453) displayed low affinity. In conclusion, S 16924 displays a profile of activity in diverse models of potential antipsychotic and extrapyramidal properties similar to that of clozapine and different to that of haloperidol. In particular, reflecting its partial agonist actions at 5-HT1A receptors, S 16924 inhibits rather than induces catalepsy in rats. However, in contrast to clozapine, S 16924 displays only low affinity for muscarinic and histaminic receptors. PMID- 9732400 TI - Potentiation of CD95L-induced apoptosis of human malignant glioma cells by topotecan involves inhibition of RNA synthesis but not changes in CD95 or CD95L protein expression. AB - Topotecan is a novel topoisomerase I inhibitor that may have a role in the adjuvant chemotherapy of several solid tumors, including malignant glioma. Here, we have characterized the time- and concentration-dependent toxicity of topotecan in four human malignant glioma cell lines, LN-18, LN-229, LN-308 and T98G. High micromolar concentrations of topotecan, which are unlikely to be achieved in plasma in human patients in vivo, were cytotoxic within 48 hr, induced DNA fragmentation, did not induce major cell cycle changes, failed to consistently alter BCL-2 or BAX protein levels but inhibited RNA synthesis and induced cleavable DNA/topoisomerase I complex formation. Prolonged exposure for 72 hr to high nanomolar to low micromolar concentrations of topotecan augmented p21 protein levels and induced G2/M arrest but failed to consistently alter BCL-2 and BAX protein levels, did not induce significant DNA/topoisomerase I complex formation and did not inhibit RNA synthesis. Neither short-term nor long-term topotecan toxicity was blocked by ectopic expression of bcl-2 or wild-type p53. Transfer of a mutant p53 gene enhanced topotecan sensitivity in wild-type p53 LN 229 but not mutant p53 LN-18 cells. CD95 ligand (CD95L)-induced apoptosis was synergistically enhanced by short-term/high concentration but not long-term/low concentration exposure to topotecan, suggesting that topotecan sensitizes human malignant glioma cells to CD95L-induced apoptosis via inhibition of RNA synthesis. These data suggest that topotecan needs to be administered in high concentrations, such as an intratumoral polymer, to limit glioma cell growth in synergy with CD95L in vivo. PMID- 9732401 TI - Role of cyclooxygenase-2 in the healing of gastric ulcers in rats. AB - To elucidate the role of cyclooxygenase (COX)-2 in ulcer healing, we compared the effects of NS-398 (COX-2-selective inhibitor) and indomethacin (nonselective COX inhibitor) on the healing of acetic acid-induced gastric ulcers in rats. Prostaglandin E2 (PGE2) production was elevated in ulcerated tissue, but remained unaffected in intact tissue. COX-2 mRNA was only detected in the ulcerated tissue, in which the COX-2 protein was found in fibroblasts, macrophages/monocytes and granulocytes. In contrast, COX-1 mRNA expression was not affected by ulceration. In an in vitro study, the increased PGE2 production was inhibited by NS-398; this had no effect on PGE2 production in the intact tissue. When NS-398 and indomethacin were administered to rats, 3 and 6 mg/kg NS 398 only reduced PGE2 production in the ulcerated tissue, but 10 mg/kg NS-398 and 0.5 to 2 mg/kg indomethacin inhibited the production in both the ulcerated and intact tissues. The healing of gastric ulcers was significantly impaired by 3 to 10 mg/kg NS-398 and 1 and 2 mg/kg indomethacin. The delay in ulcer healing was associated with the inhibition of PGE2 production in the ulcerated tissue. As observed upon histological analysis, regeneration of the mucosa, maturation of the ulcer base and angiogenesis in the base were significantly prevented by 6 mg/kg NS-398 and 2 mg/kg indomethacin, although the inhibitory effect of NS-398 was weaker than that of indomethacin. These results clearly indicate that COX-2 plays an important role in the healing of gastric ulcers in rats. PMID- 9732402 TI - Hepatic sinusoidal membrane transport of anionic drugs mediated by anion transporter Npt1. AB - The purpose of our study was to establish the localization of the anion transporter Npt1 in liver and the relevance of Npt1 to carrier-mediated hepatic transport of beta-lactam antibiotics. Immunocytochemical examination of mouse liver with antiserum for Npt1 showed basolateral (sinusoidal) membrane localization. Function of Npt1 was characterized in Xenopus laevis oocytes. Injection of in vitro-transcribed cRNA into oocytes resulted in an increased uptake of [14C]benzylpenicillin (PCG). The Npt1-mediated uptake was saturable with a Michaelis constant (Km) of 0.46 +/- 0.18 mM and a maximum rate (Vmax) of 46.6 +/- 8.5 pmol/60 min/oocyte, and the uptake of [14C]PCG was independent of Na+ and pH, but dependent on chloride ion. Npt1-mediated [14C]PCG uptake was inhibited by several beta-lactam antibiotics and probenecid. Oocytes injected with Npt1-cRNA demonstrated significantly enhanced transport activity for other anionic compounds such as [14C]faropenem, [14C]foscarnet and [3H]mevalonic acid, as well as [14C]PCG, compared with water-injected oocytes. In conclusion, Npt1 is suggested to participate in hepatic sinusoidal membrane transport of organic anions such as beta-lactam antibiotics as well as inorganic anions for the efflux from hepatocyte-to-blood direction. PMID- 9732403 TI - The effects of stress on homeostasis in JCR-LA-cp rats: the role of nitric oxide. AB - We have investigated the effects of early phases of chronic stress on generation and actions of nitric oxide (NO) in JCR:LA-cp rats both lean (+/+) and obese (cp/cp). Restraint stress was carried out for a 15-min single exposure or for 1 hr every day during 4, 9 or 14 days. The stress reaction was evidenced by significant increase in plasma cortisol. The exposure to stress for 14 days led to a neuronal damage in lean rats as evidenced by a decrease in glutamate uptake and an increase in the release of lactate in synaptosomes. This effect was not observed in obese rats. Concomitantly, the levels of glutamate increased in the hippocampus at 14 days in lean, but not obese rats, that showed higher basal levels of glutamate than lean rats. The activity of NO synthase (NOS) and guanosine cyclic monophosphate levels increased in the hippocampus preceding the neuronal damage. The neuronal lesions were prevented by inhibition of NOS without affecting cortisol levels. In the cardiovascular system, chronic stress exerted no significant effect on blood pressure, aortic contractility or platelet aggregation. However, there were significant changes in plasma nitrite/nitrate that reached maximum at 4 to 9 days. It is concluded that the generation of NO contributes to the systemic response to the organism to stress. In the brain, NO appears to be detrimental as this molecule mediates glutamate-dependent hippocampal damage, this effect being cortisol-independent. In contrast, in the vascular system, increased generation of NO may attenuate the vasoconstrictor and platelet aggregatory effects of catecholamines and other mediators of stress. PMID- 9732404 TI - Purification of two rat hepatic proteins with A-esterase activity toward chlorpyrifos-oxon and paraoxon. AB - A-esterases are calcium-dependent hydrolases that can detoxify the active metabolites (oxons) of organophosphorus insecticides such as chlorpyrifos and parathion. A-esterases from rat liver have previously been shown to hydrolyze chlorpyrifos-oxon but not paraoxon at low substrate concentrations. Two A esterases were extracted by ammonium sulfate fractionation from solubilized rat liver microsomes followed by gel filtration chromatography and preparative scale isoelectric focusing. The proteins displayed similar characteristics and were difficult to separate; both had similar high molecular mass and isoelectric point range and exhibited A-esterase activity toward high and low concentrations of chlorpyrifos-oxon and high concentrations of paraoxon. Sufficient amounts of the higher molecular mass protein were obtained for kinetic studies, which yielded a Km of 0.93 mM toward high concentrations of chlorpyrifos-oxon and a Vmax of 369 nmoles product formed/mg protein-min. The protein hydrolyzed phenyl acetate, chlorpyrifos-oxon and paraoxon, suggesting that arylesterase and A-esterase activities are attributable to the same liver protein(s). Assays of purified protein and kinetic studies of microsomes suggested that the activity toward high (320 microM) and low ( AF-DX 116 (3 nM) > pirenzepine (300 nM). Comparison of these minimum ACh releasing concentrations to the known affinities of the antagonists for the five mAChR subtypes is consistent with the conclusion that the autoreceptor regulating mPRF ACh release is the M2 subtype. Considerable evidence supports a role for cholinergic neurotransmission and postsynaptic M2 receptors in the mPRF in regulating levels of arousal. The present data suggest that presynaptic M2 receptors contribute to the regulation of arousal states by modulating mPRF ACh release. PMID- 9732411 TI - HMR 1883, a novel cardioselective inhibitor of the ATP-sensitive potassium channel. Part I: effects on cardiomyocytes, coronary flow and pancreatic beta cells. AB - The novel sulfonylthiourea HMR 1883 was investigated in in vitro systems. The rilmakalim-induced shortening of the APD90 in guinea pig right papillary muscle at pHo = 6.0 was antagonized half-maximally by glibenclamide and HMR 1883 with 0.14 microM and 0. 6 microM, respectively. Hypoxia-induced shortening of the APD90 was significantly attenuated by the sulfonylureas when applied 60 min after induction of hypoxia. In isolated guinea pig ventricular myocytes the APD90 as well as the whole-cell current was measured with the patch-clamp technique. The rilmakalim-induced shortening of the APD90 was half-maximally antagonized by glibenclamide and HMR 1883 with 10 nM and 0.4 microM, respectively (pHo = 6.5). The rilmakalim-induced whole-cell current (at 0 mV clamp-potential) was inhibited by glibenclamide and HMR 1883 half-maximally with 20 nM and 0.8 microM, respectively (pHo = 7.4). In isolated perfused guinea pig hearts, the coronary flow (CF) was increased by perfusion with hypoxic solution (20% O2). Whereas 1 microM glibenclamide completely inhibited the hypoxia-induced increase in CF, 10 microM HMR 1883 reduced it by only 18%. Pancreatic effects were investigated in rat insulinoma cells (RINm5F), which were hyperpolarized with 100 microM diazoxide. Addition of glibenclamide or HMR 1883 depolarized the cell potential half-maximally with concentrations of 9 nM and approximately 20 microM, respectively. In conclusion, the sulfonylthiourea HMR 1883 blocks KATPs in cardiac muscle cells with 10-50 fold higher potency than in pancreatic beta-cells and has little effect on the coronary vascular system. Therefore, HMR 1883 has pharmacological selectivity for cardiac myocytes and thereby may be a promising substance for the prevention of ischemia-induced ventricular fibrillation. PMID- 9732412 TI - HMR 1883, a novel cardioselective inhibitor of the ATP-sensitive potassium channel. Part II: effects on susceptibility to ventricular fibrillation induced by myocardial ischemia in conscious dogs. AB - The activation of the ATP-sensitive potassium channel (KATP) during myocardial ischemia leads to potassium efflux, reductions in action potential duration and the formation of ventricular fibrillation (VF). Drugs that inactivate KATP should prevent these changes and thereby prevent VF. However, most KATP antagonists also alter pancreatic channels, which promote insulin release and hypoglycemia. Recently, a cardioselective KATP antagonist, HMR 1883, has been developed that may offer cardioprotection without the untoward side effects of existing compounds. Therefore, VF was induced in 13 mongrel dogs with healed myocardial infarctions by a 2-min coronary artery occlusion during the last minute of a submaximal exercise test. On subsequent days, the exercise-plus-ischemia test was repeated after pretreatment with HMR 1883 (3.0 mg/kg i.v., n = 13) or glibenclamide (1.0 mg/kg i.v., n = 7). HMR 1883 (P < .001) and glibenclamide (P < .01) prevented VF in 11 of 13 and 6 of 7 animals, respectively. Glibenclamide, but not HMR 1883, elicited increases in plasma insulin and reductions in blood glucose. Glibenclamide also reduced (P < .01) both mean coronary blood flow and left ventricular dP/dt maximum as well as the reactive hyperemia induced by 15 sec coronary occlusions (-30.3 +/- 11%), whereas HMR 1883 did not alter this increase in coronary flow (-3.0 +/- 4.7%). Finally, myocardial ischemia (n = 10) significantly (P < .01) reduced refractory period (control, 121 +/- 2 msec; occlusion, 115 +/- 2 msec), which was prevented by either glibenclamide or HMR 1883. Thus, the cardioselective KATP antagonist HMR 1883 can prevent ischemically induced reductions in refractory period and VF without major hemodynamic effects or alterations in blood glucose levels. These data further suggest that the activation of KATPs may play a particularly important role in both the reductions in refractory period and lethal arrhythmia formation associated with myocardial ischemia. PMID- 9732413 TI - Prenatal exposure to fluoxetine (Prozac) produces site-specific and age-dependent alterations in brain serotonin transporters in rat progeny: evidence from autoradiographic studies. AB - The present study provides the first autoradiographic evidence of age-dependent regional changes in the density of serotonin (5-HT) transporters in offspring following prenatal exposure to fluoxetine. Pregnant rats received either saline or fluoxetine (10 mg/kg, s.c.) daily from gestational day 13 through 20. The density of [3H]citalopram-labeled 5-HT transporters was determined in forebrain regions and in midbrain raphe nuclei of prepubescent and adult male offspring. Brain regions representing integral components of the limbic system were particularly sensitive to the prenatal treatment. For example, prenatal fluoxetine exposure significantly altered the density of 5-HT transporters in subregions of the hypothalamus (dorsomedial nucleus, -21%; lateral hypothalamus, +21%), hippocampus (CA2, +47%; CA3, +38%), and amygdala (basolateral nucleus, +32%; medial nucleus, +44%) in prepubescent offspring. However, 5-HT transporter density in the dorsal and median raphe was unaltered in this same group of offspring. In adult offspring, 5-HT transporter densities, in all brain regions examined, were not significantly altered by prenatal exposure to fluoxetine. The present study also identifies significant age-related differences in 5-HT transporter densities between prepubescent and adult control offspring. For example, in adult control offspring, densities of 5-HT transporters were significantly greater in the cingulate cortex (+33%), basolateral amygdala (+58%), and CA1 area of the hippocampus (+78%); but significantly lower in the temporal cortex (-65%) and median raphe (-25%). The age-dependent and site specific alterations in the density of 5-HT transporters suggests that either 5 HT innervation and/or 5-HT neuron function in various forebrain regions may be altered by prenatal exposure to fluoxetine. PMID- 9732414 TI - Serotonin-mediated palmitoylation and depalmitoylation of G alpha proteins in rat brain cortical membranes. AB - We investigated serotonin stimulated palmitoylation of G alpha subunits in rat brain cerebrocortical membranes. Serotonin dose dependently stimulated palmitoylation of membrane G alpha proteins. The highest [3H] palmitate incorporation observed was by G alpha-q (7-fold), followed by G alpha-o (5-fold), G alpha-i (4-fold) and G alpha-s (3-fold) and these increases in palmitoylation were blocked by methiothipin, a serotonin receptor antagonist. Isoproterenol selectively stimulated G alpha-s palmitoylation which was blocked by propranalol. Immunoprecipitates of palmitoylated G alpha subunits yielded single labeled bands on SDS-PAGE. In an attempt to define the sequence of palmitoylation/depalmitoylation that follows receptor stimulation, nonreceptor mediated palmitoylation was carried out in the presence of guanine nucleotides and receptor mediated G alpha depalmitoylation was then monitored. Receptor stimulation did not result in depalmitoylation when membranes were prelabeled with [3H] palmitic acid in the presence of the nonhydrolyzable analogue of GTP, Gpp(NH)p. However, serotonin receptor stimulation in the presence of guanine nucleotides, depalmitoylated (90%) membrane G alpha proteins when prelabeled in the presence of GTP. Coimmunoprecipitation experiments revealed decrease in G beta immunoreactivity associated with G alpha immunoprecipitates obtained from membranes prelabeled in presence of GTP prior to reincubation with Gpp(NH)p and serotonin. These observations suggest that receptor occupation results in depalmitoylation of the trimer, followed by guanine nucleotide exchange and dissociation of the alpha subunit from beta-gamma dimer and that the activated alpha subunit is a substrate for repalmitoylation. PMID- 9732416 TI - Evaluation of expanded polytetrafluoroethylene arteriovenous access grafts onto which microvessel-derived cells were transplanted to "improve" graft performance: preliminary results. AB - The purpose of this study was to implement and evaluate a clinical protocol for following longitudinally the luminal responses of microvessel cell seeded expanded polytetrafluoroethylene (ePTFE) vascular grafts implanted for hemodialysis access. Half of the patients enrolled in the study were randomized to receive grafts that were "seeded" with transplanted microvessel cells derived from autologous subcutaneous fat; the other half of the patients received nonseeded grafts. The patients agreed to scheduled biopsies of their grafts at three postoperative times. All biopsy samples were evaluated by routine histologic and electron microscopy techniques. Three men and six women were enrolled in the study. All operative procedures were tolerated well. However, only two of the nine patients agreed to 1-year postimplantation biopsies; one of these patients had been randomized to receive a "nonseeded" ePTFE graft and one randomized to receive a "seeded" graft. The "seeded" graft at 3 months showed endothelial cells on the luminal surface as well as some intimal thickening. By 20 months, the same "seeded" graft showed significant concentric intimal thickening and by 24 months, this "seeded" graft thrombosed. The "nonseeded" graft at 16 months had irregular areas of intimal thickening which were quite patchy in nature. The flow contacting surface of the "nonseeded" graft remained thin. The intima of the "seeded" graft was twice as thick as that of the "nonseeded" graft. The methodologies implemented in the study design were appropriate. Biopsy samples were obtained without complication and were easily processed for analysis. Patient compliance with the biopsy protocol was problematic however. The study was terminated because of the development of significant concentric intimal hyperplasia in a "seeded" graft. PMID- 9732415 TI - Identification of new human CYP2C19 alleles (CYP2C19*6 and CYP2C19*2B) in a Caucasian poor metabolizer of mephenytoin. AB - A genetic polymorphism in the metabolism of the anticonvulsant drug S-mephenytoin has been attributed to defective CYP2C19 alleles. This genetic polymorphism displays large interracial differences with the poor metabolizer (PM) phenotype representing 2-5% of Caucasian and 13-23% of Oriental populations. In the present study, we identified two new mutations in CYP2C19 in a single Swiss Caucasian PM outlier (JOB 1) whose apparent genotype (CYP2C19*1/CYP2C19*2) did not agree with his PM phenotype. These mutations consisted of a single base pair mutation (G395A) in exon 3 resulting in an Arg132-->Gln coding change and a (G276C) mutation in exon 2 resulting in a coding change Glu92-->Asp. However, the G276C mutation and the G395A mutation resided on separate alleles. Genotyping tests of a family study of JOB1 showed that the exon 2 change occurred on the CYP2C19*2 allele, which also contained the known splice mutation in exon 5 (this variant is termed CYP2C19*2B to distinguish it from the original splice variant now termed CYP2C19*2A). The exon 3 mutation resided on a separate allele (termed CYP2C19*6). In all other respects this allele was identical to one of two wild-type alleles, CYP2C19*1B. The incidence of CYP2C19*6 in a European Caucasian population phenotyped for mephenytoin metabolism was 0/344 (99% confidence limits of 0 to 0.9%). Seven of 46 Caucasian CYP2C19*2 alleles were CYP2C19*2B(15%) and 85% were CYP2C19*2A. The Arg132Gln mutation was produced by site-directed mutatgenesis and the recombinant protein expressed in a bacterial cDNA expression system. Recombinant CYP2C19 6 had negligible catalytic activity toward S-mephenytoin compared with CYP2C19 1B, which is consistent with the conclusion that CYP2C19*6 represents a PM allele. Thus, the new CYP2C19*6 allele contributes to the PM phenotype in Caucasians. PMID- 9732418 TI - The effect of hypercholesterolemia on the rabbit transarterial wall oxygen gradient. AB - The objective of this study was to determine the effect of hypercholesterolemia on the transarterial wall oxygen gradient. Female New Zealand white rabbits (3-4 kg) were fed a 0.5% cholesterol supplemented diet or a 0.25% cholesterol supplemented diet and their transarterial wall oxygen gradients measured prior to the formation of atherosclerotic lesions at 4 weeks (0.5% cholesterol group) or 8 weeks (0.25% cholesterol diet) after beginning the diet. Arterial blood oxygen content and arterial blood pressure were recorded during the experiments. Control rabbits had a serum cholesterol level of 52.8 +/- 6 mg/dl, rabbits fed the 0.25% cholesterol diet had serum cholesterol levels of 579.5 +/- 29.2 mg/dl, and those fed the 0.5% cholesterol diet had serum cholesterol levels of 1235.4 +/- 37.6 mg/dl. There was no difference in the transarterial wall oxygen gradients between any of the groups. These results were noted with no differences in arterial blood oxygen content, arterial blood pressure, or evidence of atherosclerotic lesions. Hypercholesterolemia does not alter the delivery of oxygen to the artery wall prior to the formation of atherosclerotic lesions. PMID- 9732417 TI - Locally applied antisense oligonucleotide to proliferating cell nuclear antigen inhibits intimal thickening in experimental vein grafts. AB - This study examines the effect of antisense oligonucleotide to proliferating cell nuclear antigen (PCNA) on the formation of vein graft intimal hyperplasia in vivo, using localized administration. Twenty-four New Zealand white rabbits had a right carotid interposition bypass graft using the external jugular vein and were sacrificed on the 28th postoperative day. To determine the effect of PCNA on the development of intimal hyperplasia, 6 animals had their grafts coated with a pluronic gel containing 18 base antisense oligonucleotide to PCNA (1 mg/ml), 6 received a pluronic gel containing an 18 base nonsense oligonucleotide (1 mg/ml), and 12 animals were controls (6 with and 6 without pluronic gel). These grafts were harvested for morphology and videomorphometry. There was no change in the intimal thickness between the control and gel-treated groups. (70 +/- 4 microm versus 72 +/- 4 microm; mean +/- s.e.m.; p = ns). The presence of nonsense oligonucleotide had no further effect. Antisense PCNA produced a 26% decrease in intimal thickness to 50 +/- 4 microm in the treated vein grafts (p < 0.03) without a change in medial thickness. This study shows that a local single application of antisense oligonucleotide to PCNA will reduce the intimal hyperplasia in experimental vein grafts over 28 days. PMID- 9732419 TI - Chronic loading and extension increases the acute breaking strength of polypropylene sutures. AB - Polypropylene sutures provide satisfactory strength for construction of vascular anastomoses, but occasionally they break. Experimental studies show that they break at reduced forces when they are subjected to chronic loads. Moreover, in patients, sutures are subject to acute loads superimposed on chronic loads. For example, an episode of hypertension applies acute load that is added to the baseline chronic load in a suture that has been used to close an arteriotomy. The purpose of the present study was to examine the breaking force of 6-0 polypropylene sutures subjected to acute loads after they had been loaded with chronic loads. One hundred sixty-five 6-0 polypropylene sutures were subjected to 50-175 g chronic loads in vitro. After 38 days they were subjected to additional increasing acute loads until they broke. Five hundred ninety other sutures were subjected to "injuries" of manipulation before chronic loading. A stray knot was simulated by placing a knot in the center of 90 sutures. Nurse's tugs used to straighten folded sutures in the operating room were simulated by applying brief loads of 75-275 g to 452 other sutures. Intraoperative injuries were simulated in 48 other sutures by pinching them with DeBakey forceps. Surprisingly, chronic loading of polypropylene sutures increased their acute breaking force. It is suggested that this may have resulted from increased orientation of crystals in the core of the filaments. By contrast, disturbing the outer surface of the filament by placing a stray knot, or pinching with forceps decreased acute breaking strength. These data suggest that if polypropylene sutures do not break soon after they have been placed in a patient, they may gain strength over time. PMID- 9732420 TI - Carotid atherosclerosis in patients operated for lower extremity ischemia before the age of 50: a case control study. AB - The purpose of this study was to investigate the presence of, and to identify factors associated with carotid atherosclerosis in patients, previously operated on for lower extremity ischemia before the age of 50. Forty-eight patients were compared to sex- and age-matched controls. All subjects were examined with duplex ultrasonography of the neck arteries and analysis of serum lipoproteins. History including smoking habits, family history of cardiovascular disease, and medication was also obtained. The patients were examined clinically and their preoperative angiograms were reevaluated. Thirty-one patients (64%) and 13 controls (23%) had a carotid lesion (p < 0.0001). Patients with suprainguinal or multilevel disease had a higher proportion of carotid lesions than those with only infrainguinal disease in whom the proportion was similar to the controls. A multiple regression analysis among the patients revealed that age, level of lower extremity arterial disease, presence of family history, and the ratio apolipoproteinB/apolipoproteinA discriminated significantly between those with and without carotid disease. It is concluded that a high proportion of patients operated on for lower extremity suprainguinal arterial occlusive disease at an early age have carotid lesions at follow-up, while patients operated on due to isolated infrainguinal disease have a prevalence similar to controls. PMID- 9732421 TI - 25-year trends in resection of abdominal aortic aneurysms. AB - An attempt was made to document trends that have occured over a 25-year period in clinical presentation, preoperative evaluation, operative management, and patient outcome in patients with an abdominal aortic aneurysm. The experience (574 aneurysmectomies) of one cardiovascular surgical group was analyzed by retrospective review of hospital and office records. Changes over time of patients' ages, aneurysm sizes and statuses, prior myocardial revascularization, operative mortality, and certain other parameters were evaluated. During the period of study, there was a significant decrease in aneurysm size, increase in patients' ages, and an increased incidence of previous coronary artery bypass. No ruptured aneurysm was < 5 cm in diameter. The incidence of rupture and the operative mortality in patients with a ruptured aneurysm did not change significantly. There was a significantly (p = 0.03) lower operative mortality of 0.4% in the latter half of the series for elective aneurysmectomy. Increased utilization of preoperative cardiologic evaluation, and myocardial revascularization, has been associated with a decreased operative mortality in patients undergoing elective aneurysmectomy even though the patients are now older and have more age-related comorbidities. Elective aneurysmectomy should be offered to most patients when an abdominal aortic aneurysm is > or =5 cm in diameter. PMID- 9732422 TI - Preoperative color flow Doppler imaging for fibula free tissue transfers. AB - Fibula osteocutaneous free tissue transfer to reconstruct the oromandibular complex is a widely recommended technique following oncologic resection. Preoperative determination of adequate perfusion to the donor extremity is necessary to assure lower extremity viability after flap harvest. Vascular variations and/or peripheral arterial occlusive disease (PAOD) may exist whereby sacrifice of peroneal vessels can cause ischemia to the lower leg and foot. Additionally, variability of cutaneous perforators can make the fibula skin paddle viability unpredictable. Color flow Doppler (CFD) is a reliable modality to preoperatively assess the lower extremity in fibula osteocutaneous free tissue transfer patients. Prospective CFD examination of 38 consecutive patients (76 legs) considered for fibula free flap reconstruction was performed. A standard protocol was designed to evaluate the lower extremity vasculature and identify cutaneous perforators with CFD. Findings were studied with respect to flap choice, operative findings, and reconstruction outcomes. Number of cutaneous perforators and their impact on skin paddle design were also recorded. Color flow Doppler's ability to image peroneal vessels as well as determine collateral and distal perfusion were effective. CFD accurately identified bilateral vascular anomalies in one patient (2.6%), and significant arterial disease in three patients (7.9%). Cutaneous perforators were also accurately mapped and confirmed intraoperatively in 31 patients. In several instances, the information provided by the CFD examination altered flap selection, 4/38 patients (10.5%), or skin paddle design, 5/32 patients (15.6%). Color flow Doppler allowed successful fibula transfer in all the free flap candidates with normal exams. It has the advantages of low cost and no morbidity. CFD allows for accurate mapping of fibula cutaneous perforators which facilitates skin paddle design. We recommended the use of preoperative CFD in all patients being considered for fibular free flap surgery. PMID- 9732423 TI - Computerized tomographic angiography scan following carotid endarterectomy. AB - The purpose of this study was to evaluate the role of computed tomographic angiography (CTA) for postoperative assessment of carotid endarterectomy (CE). Twenty carotid endarterectomies were performed and controlled by using (1) intraoperative angiography, (2) postoperative duplex scanning and CTA with multiprojection volume reconstruction (MPVR). Intraoperative angiographic controls were deemed satisfactory for all patients. In 12 patients, the postoperative morphological aspect was satisfactory with CTA and duplex scanning. In the eight remaining patients, CTA and/or duplex scanning revealed 12 abnormalities: 3 were equally visualized on CTA and duplex scanning, 6 only on CTA and 3 only on duplex scanning. CTA is a rapid and noninvasive technique allowing the surgeon to get informative and comparative data. It might be an interesting alternative to postoperative angiography. PMID- 9732424 TI - Fresh and cryopreserved arterial homografts in the treatment of prosthetic graft infections: experience of the Italian Collaborative Vascular Homograft Group. AB - Following the experience of cardiac surgeons with homografts in the treatment of infective aortic valve endocarditis, cardiovascular surgeons have investigated in situ revascularization by means of homografts in the management of vascular prosthetic graft infections. Preliminary results are encouraging, but their late fate in long-term follow-up and the influence of preservation techniques are still under investigation. This article reports the experience of the Italian Collaborative Vascular Homograft Group, with the use of fresh and cryopreserved arterial homografts for the treatment of prosthetic graft infections. Between March 1994 and December 1996, 44 patients with prosthetic graft infection were treated with homografts (13 preserved at 4 degrees C, 31 cryopreserved). The mean age of the patients was 65 years. Emergency surgical procedures were performed in eight patients (18%). Sepsis was diagnosed in 11 patients, aortoenteric fistula in 13, and false aneurysms in 10. Staphylococcus was the main cause of infection. The types of vascular reconstruction with homograft were: 32 aortobifemoral, 3 aortoaortic, 2 iliofemoral, 4 peripheral, and 3 axillobifemoral. Human lymphocyte antigen (HLA) and antibody (ABO) blood group system compatibility between donors and recipients was not respected. The mean duration of follow-up was 15 months (range 1-33). Clinical and duplex scanning evaluations were routinely performed. Computed tomography (CT) or magnetic resonance (MR) scanning or arteriography were performed on the basis of duplex scanning results. There were six deaths during the early postoperative period (30 days) with a mortality rate of 13.6%. During the follow-up there were five late deaths with a mortality rate of 11.4%. Eight patients had graft occlusion. Three cases were successfully treated with thrombectomy. Two cases were successfully treated with femoropopliteal bypass with autologous vein. In three cases leg amputation was necessary. The results of fresh and cryopreserved homograft were compared. No significative differences of early postoperative mortality, late mortality, homograft related mortality, and graft occlusion were observed. We have evaluated the actuarial survival of the patients and the actuarial patency of the homografts on the aortoiliac reconstructions. Twelve months after the surgery the actuarial survival of the patients was 73% and the actuarial patency of the homografts was 56%. In our preliminary experience, we have not observed any significant difference in terms of clinical outcome by using fresh rather than cryopreserved homografts. In the near future it will be our policy to employ only cryopreserved homografts. Moreover, we will extend vessel harvesting to nonheart-beating donors, thus maximizing retrieval. The aforementioned solutions will supply the best graft availability to obtain dimensional and ABO compatibility between donors and recipients. PMID- 9732425 TI - Internal carotid artery dissection in a patient with Behcet's syndrome. AB - The authors report a case of internal carotid artery dissection in a young woman with Behcet's syndrome. The authors postulate that a vasculitis of the vasa vasorum already suspected as the basis of aneurysm formation in course of Behcet's syndrome can account for occurrence of arterial dissection in this inflammatory condition. PMID- 9732426 TI - Ureteropelvic urinary extravasation due to iliac artery aneurysm. AB - We report a case of left-sided hydronephrosis and ureteropelvic urinary extravasation due to a large left iliac artery aneurysm. Urinoma was diagnosed preoperatively by contrast-enhanced computed tomography. The patient was successfully treated by percutaneous nephrostomy and ureteral double J stent placement followed by staged operative repair. PMID- 9732427 TI - Catheter-directed thrombolysis in the treatment of phlegmasia cerulea dolens. AB - Phlegmasia cerulea dolens is a potentially devastating complication of extensive deep venous thrombosis for which there is currently no consensus for treatment. Heparin anticoagulation, surgical thrombectomy, thrombolytic therapy, fasciotomy, and amputation have each been advocated. We present two cases of phlegmasia cerulea dolens successfully treated with catheter-directed venous thrombolytic therapy. PMID- 9732428 TI - Cocaine-induced thrombosis of common iliac and popliteal arteries. AB - Cocaine-induced thrombosis has been reported in the literature; however, its mechanism is not fully understood. Most cases are of small caliber vessels, such as the coronaries and cerebral vasculature. We report a case of a 36-year-old man with signs and symptoms of acute arterial insufficiency in his right lower extremity. At angiography, the right common iliac artery and the popliteal artery were occluded. The patient was successfully treated with thrombolytic therapy. Cocaine-induced thrombosis should be suspected in a patient with history of cocaine abuse who presents with acute arterial insufficiency in an extremity, without an identifiable source. PMID- 9732430 TI - Animal models for the study of lower extremity chronic venous disease: lessons learned and future needs. AB - The purpose of this review is to define animal models of chronic venous disease and to demonstrate how animal studies can impact our understanding and treatment of this disorder. To this end an extensive literature search was conducted highlighting potential animal models of chronic lower extremity venous disease. Scientific investigations using animals to study particular aspects of this disease are also reviewed. This review was conducted by members of the Committee on Research of the American Venous Forum to help provide direction for future venous research endeavors. Useful models of chronic venous occlusive disease involve controlled ligation of a major lower limb vein and multiple tributaries. Such a model can provide sustained venous hypertension and studies using this model have confirmed that an isodiametric graft can provide early hemodynamic relief. Models of primary, postphlebitic, and isolated chronic deep venous insufficiency are available for study. Valve repair or transplantation can positively impact the insufficiency observed in these models. Investigations into valve substitutes have generally been disappointing or are undergoing early evaluation. In conclusion, animal models for the study of some aspects of chronic venous disease do exist and have already affected our clinical approach to patients. The scientific study of basic pathophysiology, diagnostics, end-organ response, and long-term surgical treatments of this disorder in well-controlled animal experiments have not been conducted. PMID- 9732429 TI - May-Thurner syndrome: management by endovascular surgical techniques. AB - May-Thurner syndrome is a condition in which there is impaired venous return due to compression of the left common iliac vein by the overlying right common iliac artery. The impedance of flow occurs both by the compressive force of the artery against the vein and by resultant intraluminal web formation inside the vein. Because of the mechanical nature of the obstruction, conservative management of these patients has resulted in poor outcomes. Typically, operative therapy is recommended and perused via various approaches. We have recently treated a 15 year-old patient with this disorder utilizing endovascular surgical techniques with an excellent outcome. A 1-year follow-up is presented. PMID- 9732431 TI - Irradiation for the treatment of intimal hyperplasia. AB - Intimal hyperplasia represents a serious complication limiting the long-term benefits of vascular interventions such as balloon angioplasty and stent placement. Although pharmacological interventions have attempted to curtail restenosis, they have not been shown to be effective to date. Radiotherapy is one alternative that has shown promise as an inhibitor of intimal hyperplasia in several animal models. Irradiation causes cell death by producing irreparable damage to DNA. This is believed to be the mechanism of inhibition of VSMC proliferation. Delivery of irradiation can be either intraluminal via an angiographically directed catheter or extraluminal using an external radiation source such as an x-ray device. Intraluminal irradiation has generally utilized either gamma or beta-emitting sources. Both have been effective in producing a dose response, although some studies advocate the use of beta-type irradiation as a safer, more efficient means of delivery. Extraluminal irradiation also has been an effective inhibitor of intimal hyperplasia. Studies suggest that this form of irradiation provides a more even-dose distribution to vessel walls than an intravascular delivery system. The use of radiotherapy has more recently been extended to clinical trials, and initial studies have shown promising results. The success of irradiation must be balanced with its potential complications including radiation-induced arteritis, coronary artery stenosis, and secondary development of malignancy. Although these have been associated with irradiation, the dose used in these cases was often considerably higher than those used in the treatment of intimal hyperplasia. Finally, with the advent of gene therapy, irradiation may provide an additional means of supplementing this new type of therapy through radiation-inducible gene therapy. PMID- 9732432 TI - Surgical management of chronic venous insufficiency. PMID- 9732433 TI - Thoracoscopic sympathectomy. PMID- 9732435 TI - Tetramethylammonium:coenzyme M methyltransferase system from methanococcoides sp AB - A methanogen (strain NaT1) that belongs to the family of Methanosarcinaceae and that can grow on tetramethylammonium as the sole energy source has recently been isolated. We report here that cell extracts of the archaeon catalyze the formation of methyl-coenzyme M from coenzyme M and tetramethylammonium. The activity was dependent on the presence of Ti(III) citrate and ATP, and was rapidly lost under oxic conditions. Anoxic chromatography on DEAE-Sepharose revealed that two fractions, fractions 3 and 4, were required for activity. A 50 kDa protein that together with fraction 3 catalyzed methyl-coenzyme M formation from tetramethylammonium and coenzyme M was purified from fraction 4. From fraction 3, a 22-kDa corrinoid protein and a 40-kDa protein exhibiting methylcobalamin:coenzyme M methyltransferase (MT2) activity were purified. The N terminal amino acid sequences of these purified proteins were determined. The 40 kDa protein showed sequence similarity to MT2 isoenzymes from Methanosarcina barkeri. Cell extract of strain NaT1 grown on trimethylamine rather than on tetramethylammonium did not exhibit tetramethylammonium:coenzyme M methyltransferase activity. The strain was identified as belonging to the genus of Methanococcoides, its closest relative being Methanococcoides methylutens. PMID- 9732434 TI - Methylglyoxal production in bacteria: suicide or survival? AB - Methylglyoxal is a toxic electrophile. In Escherichia coli cells, the principal route of methylglyoxal production is from dihydroxyacetone phosphate by the action of methylglyoxal synthase. The toxicity of methylglyoxal is believed to be due to its ability to interact with the nucleophilic centres of macromolecules such as DNA. Bacteria possess an array of detoxification pathways for methylglyoxal. In E. coli, glutathione-based detoxification is central to survival of exposure to methylglyoxal. The glutathione-dependent glyoxalase I-II pathway is the primary route of methylglyoxal detoxification, and the glutathione conjugates formed can activate the KefB and KefC potassium channels. The activation of these channels leads to a lowering of the intracellular pH of the bacterial cell, which protects against the toxic effects of electrophiles. In addition to the KefB and KefC systems, E. coli cells are equipped with a number of independent protective mechanisms whose purpose appears to be directed at ensuring the integrity of the DNA. A model of how these protective mechanisms function will be presented. The production of methylglyoxal by cells is a paradox that can be resolved by assigning an important role in adaptation to conditions of nutrient imbalance. Analysis of a methylglyoxal synthase-deficient mutant provides evidence that methylglyoxal production is required to allow growth under certain environmental conditions. The production of methylglyoxal may represent a high-risk strategy that facilitates adaptation, but which on failure leads to cell death. New strategies for antibacterial therapy may be based on undermining the detoxification and defence mechanisms coupled with deregulation of methylglyoxal synthesis. PMID- 9732436 TI - Nucleoids and coated vesicles of "Epulopiscium" spp AB - We describe here aspects of the anatomy of two "Epulopiscium" morphotypes, unusually large bacteria that are not yet cultured and that reproduce by the internal generation of two or more vegetative daughter cells. Two morphotypes, A and B, which are enteric symbionts of several species of herbivorous surgeonfish (Acanthuridae), were collected around the Great Barrier Reef of Australia, preserved there, and later stained for light microscopy. Some samples were examined by electron microscopy. In both morphotypes, countless discrete nucleoplasms or nucleoids were found to occupy a single shallow layer just beneath the surface all around these organisms. At each end of the morphotype B cells, a membrane-bound compartment containing dense cords of chromatin was observed. When these were found at each end of growing daughter cells, no polar compartments were then found in their mother organism. Electron micrographs of sections of morphotype A symbionts show that their outermost region is composed of tightly packed coated vesicles, each surrounded by a thin, dense, spacious capsule. Near the surface of type A organisms the remains of broken vesicles, broken capsules, and a finely fibrous matrix fuse to form a fabric that serves as the cell wall. Morphotype B organisms, however, were observed to have a distinct, morphologically continuous outer wall. PMID- 9732437 TI - The Serratia marcescens hemolysin is secreted but not activated by stable protoplast-type L-forms of Proteus mirabilis. AB - The outer-membrane protein ShlB of Serratia marcescens activates and secretes hemolytic ShlA into the culture medium. Without ShlB, inactive ShlA (termed ShlA*) remains in the periplasm. Since Proteus mirabilis L-form cells lack an outer membrane and a periplasm, it was of interest to determine in which compartment recombinant ShlA* and ShlB are localized and whether ShlB activates ShlA*. The cloned shlB and shlA genes were transcribed in P. mirabilis stable L form cells by the temperature-inducible phage T7 RNA polymerase. Radiolabeling, Western blotting, and complementation with C-terminally truncated ShlA (ShlA255) identified inactive ShlA* in the culture supernatant. ShlB remained cell-bound and did not activate ShlA without integration in an outer membrane. Although hemolytic ShlA added to L-form cells had access to the cytoplasmic membrane, it did not affect L-form cells. Synthesis of the large ShlA protein (165 kDa) in P. mirabilis L-form cells under phage T7 promoter control demonstrates that L-form cells are suitable for the synthesis and secretion of large recombinant proteins. This property and the easy isolation of released proteins make L-form cells suitable for the biotechnological production of proteins. PMID- 9732438 TI - High genetic and physiological diversity of sulfate-reducing bacteria isolated from an oligotrophic lake sediment. AB - The community structure of sulfate-reducing bacteria in littoral and profundal sediments of the oligotrophic Lake Stechlin (Germany) was investigated. A collection of 32 strains was isolated from the highest positive dilutions of most probable-number series, and their partial 16S rRNA gene sequences and genomic fingerprints based on ERIC (enterobacterial repetitive intergenic consensus)-PCR were analyzed. The strains fell into eight distinct phylogenetic lineages, and the majority (70%) showed a close affiliation to the genus Desulfovibrio. Most of the remaining strains (22%) were related to the gram-positive Sporomusa and Desulfotomaculum groups. A high redundancy of 16S rRNA gene sequences was found within several of the phylogenetic lineages. This low phylogenetic diversity was most pronounced for the subset of strains isolated from oxic sediment layers. ERIC-PCR revealed that most of the strains with identical 16S rRNA gene sequences were genetically different. Since strains with identical 16S rRNA gene sequences but different genomic fingerprints also differed considerably with respect to their physiological capabilities, the high diversity detected in the present work is very likely of ecological relevance. Our results indicate that a high diversity of sulfate-reducing bacterial strains can be recovered from the natural environment using the established cultivation media. PMID- 9732439 TI - Environmental and physiological factors affecting the uptake of phosphate by chlorobium limicola AB - The uptake of soluble phosphate by the green sulfur bacterium Chlorobium limicola UdG6040 was studied in batch culture and in continuous cultures operating at dilution rates of 0.042 or 0.064 h-1. At higher dilution rates, washout occurred at phosphate concentrations below 7.1 &mgr;M. This concentration was reduced to 5. 1 &mgr;M when lower dilution rates were used. The saturation constant for growth on phosphate (K&mgr;) was between 2.8 and 3.7 &mgr;M. The specific rates of phosphate uptake in continuous culture were fitted to a hyperbolic saturation model and yielded a maximum rate (Vamax) of 66 nmol P (mg protein)-1 h-1 and a saturation constant for transport (Kt) of 1.6 &mgr;M. In batch cultures specific rates of phosphate uptake up to 144 nmol P (mg protein)-1 h-1 were measured. This indicates a difference between the potential transport of cells and the utilization of soluble phosphate for growth, which results in a significant change in the specific phosphorus content. The phosphorus accumulated within the cells ranged from 0.4 to 1.1 &mgr;mol P (mg protein)-1 depending on the growth conditions and the availability of external phosphate. Transport rates of phosphate increased in response to sudden increases in soluble phosphate, even in exponentially growing cultures. This is interpreted as an advantage that enables Chl. limicola to thrive in changing environments. PMID- 9732441 TI - Identification of phototrophic sulfur bacteria through the analysis of lmwRNA band patterns AB - Several phototrophic sulfur bacteria were identified preliminarily through the analysis of the low-molecular-weight RNA fraction (lmwRNA) of bacterial cells. This fraction includes the ribosomal 5S RNA and several transfer RNAs. These molecules were separated by high-resolution electrophoresis in polyacrylamide gels, and the resulting band patterns were used as fingerprints for the identification of the organisms. We examined a large number of well-characterized reference strains together with a broad range of purple sulfur bacterial isolates from freshwater and marine environments. A cluster analysis was run using the similarity matrix calculated from the band patterns. Despite the shortcomings of the method, close relatives were clustered together yielding a number of groups consistent with the phylogenetic arrangement established through the analyses of a few available 16S rRNA gene sequences. Thus, the classification obtained gives further support to rearrangement of the group as the analyses of 16S rRNA gene sequences had previously suggested. We conclude that the analysis of lmwRNA band patterns is a rapid and simple tool for grouping and preliminarily identifying new isolates of phototrophic sulfur bacteria. PMID- 9732440 TI - Mutations in cell division proteins FtsZ and FtsA inhibit phiX174 protein-E mediated lysis of Escherichia coli. AB - Electron microscopic studies emphasized that the protein-E-specific transmembrane tunnel structure, which permeabilizes Escherichia coli, is not randomly distributed over the cell envelope but is restricted to areas of potential division sites. These sites were located predominantly in the middle of the cell, but approximately one-third of these structures are found at the polar sites. Therefore, E. coli mutant strains with defects in cell division components were tested for their sensitivity to protein-E-mediated lysis. The ftsZ84 and the ftsA12 cell division mutant strains of E. coli were tolerant to protein-E mediated lysis, whereas the ftsA3 mutant strain was lysed by protein E under conditions nonpermissive for division. The protein-E-tolerant phenotype of ftsZ84 and ftsA12 and the lysis-sensitive phenotype of other components of the septosome (e.g., ftsA3, ftsQ, and ftsI) suggest that initiation of cell division - rather than specific functions of cell division - plays an essential role in protein-E mediated lysis. SulA-overproducing cells had a lysis-positive phenotype, the ring structure - but not the GTPase function - of FtsZ was impaired. PMID- 9732442 TI - Metronidazole resistance and microaerophily in Campylobacter species. AB - Metronidazole is active against most anaerobic organisms and is also used in the treatment of the microaerophilic bacterium Helicobacter pylori. Resistance to metronidazole is uncommon in most anaerobic organisms, but it is increasingly prevalent in H. pylori. Previously we have suggested that metronidazole resistance in H. pylori is inherent in the microaerophilic nature of the organism and therefore would be present in other microaerophiles such as Campylobacter. Short periods of anaerobiosis caused metronidazole-resistant (MtrR) strains of Campylobacter spp. to become sensitive to metronidazole. Under microaerophilic conditions, cultures of the MtrR mutant Campylobacter coli R1 at bacterial cell densities of greater than 10(8) cfu/ml lost viability, whereas no loss in viability was observed in cultures at cell densities of less than 10(8). The MtrS C. coli strain lost viability at all cell densities. Comparisons of NAD(P)H oxidase activity between MtrS and MtrR strains indicated that the MtrS C. coli strain contained fourfold higher NADH oxidase activity and twofold higher NADPH oxidase activity than did the MtrR Campylobacter strains. These results show that MtrR Campylobacter spp. display resistance characteristics similar to those of H. pylori, suggesting that the resistance mechanism is a phenomenon of the microaerophilic nature of these bacteria. PMID- 9732443 TI - Characterization of the group 1 and group 2 sigma factors of the green sulfur bacterium Chlorobium tepidum and the green non-sulfur bacterium Chloroflexus aurantiacus. AB - The group 1 and group 2 sigma70-type sigma factors of the green sulfur bacterium Chlorobium tepidum and of the green nonsulfur bacterium Chloroflexus aurantiacus were cloned and characterized. Cb. tepidum was found to contain one sigma70-type sigma factor; the expression of the gene was analyzed by Northern blot hybridization and primer-extension mapping. Cf. aurantiacus has genes encoding four sigma factors of groups 1 and 2. The expression of these genes was examined in cells grown aerobically and anaerobically. The sigC gene was expressed at approximately equal levels under both conditions, resulting in its designation as the group 1 sigma factor of this organism. The only other detectable transcripts arose from the sigB gene, which was expressed at higher levels during aerobic growth. A phylogenetic tree was obtained using the group 1 sigma factors of Cb. tepidum, Cf. aurantiacus, and diverse eubacteria as the molecular marker. The resulting phylogenetic tree shows that Cb. tepidum and Cf. aurantiacus are related to each other and to the cyanobacteria. The relationship of the group 2 sigma factors of Cf. aurantiacus and the cyanobacteria was more specifically examined phylogenetically. The group 2 sigma factors of Cf. aurantiacus probably arose by gene duplication events after the split of the green nonsulfur bacteria from other photosynthetic eubacteria. PMID- 9732444 TI - Membrane-associated redox activities in Thermotoga neapolitana. AB - Elemental sulfur reduction by the hyperthermophilic bacterium Thermotoga neapolitana provides an alternative to hydrogen evolution during fermentation. Electrons are transferred from reduced cofactors (ferredoxin and NADH) to sulfur by a series of unknown steps. One enzyme that may be involved is an NADH:methyl viologen oxidoreductase (NMOR), an activity that in other fermenting organisms is associated with NADH:ferredoxin oxidoreductase. We found that 83% of NMOR activity was contained in the pellet fraction of cell extracts subjected to ultracentrifugation. This pellet fraction, presumably containing cell membranes, was required for electron transfer to NAD+ from ferredoxin-dependent pyruvate oxidation. However, the NMOR activity in this fraction used neither Thermotoga nor clostridial ferredoxins as substrates. NMOR activity was also detected in aerobically prepared vesicles. By comparison with ATPase activities, NMOR was found primarily on the cytoplasmic face of these vesicles. During these studies, an extracytoplasmic hydrogenase activity was discovered. In contrast to the soluble hydrogenase, this hydrogenase activity was completely inhibited when intact cells were treated with cupric chloride and was present on the extracytoplasmic face of vescides. In contrast to a soluble hydrogenase reported in Thermotoga maritima, this activity was air-stable and was inhibited by low concentrations of nitrite. PMID- 9732445 TI - Expression of the succinate dehydrogenase genes (sdhCAB) from the facultatively anaerobic paenibacillus macerans during aerobic growth AB - Paenibacillus (formerly Bacillus) macerans is capable of succinate oxidation under oxic conditions and fumarate reduction under anoxic conditions. The reactions are catalyzed by different enzymes, succinate dehydrogenase (Sdh) and fumarate reductase (Frd). The genes encoding Sdh (sdhCAB) were analyzed. The gene products of sdhA and sdhB were similar to the subunits of known Sdh and Frd enzymes. The hydrophobic subunit SdhC showed close sequence similarity to the class of Sdh/Frd enzymes containing diheme cytochrome b. From the sdhCAB gene cluster two transcripts were produced, one comprising sdhCAB, the other sdhAB. The transcripts were found only during aerobic growth, and the amount was directly proportional to Sdh activity, but inversely proportional to Frd activity. PMID- 9732446 TI - Effects of pisatin on Dictyostelium discoideum: its relationship to inducible resistance to nystatin and extension to other isoflavonoid phytoalexins. AB - Dictyostelium discoideum amoebae can acquire resistance to otherwise inhibitory concentrations of pisatin, an isoflavonoid phytoalexin of pea, and nystatin, a polyene antibiotic, following pretreatment with sublethal concentrations of these compounds. Additionally, growth on medium containing pisatin can induce nystatin resistance. We show here that distinct mechanisms mediate the inducible resistance to these two compounds because it is possible to isolate mutations that specifically block the induction of nystatin resistance but do not affect the induction of pisatin resistance. Pisatin did not affect wild-type sterol biosynthesis; therefore, the induction of nystatin resistance by pisatin is probably not via an alteration of membrane sterols. The inducible pisatin resistance phenotype was shown to extend to the isoflavonoid phytoalexins maackiain and biochanin A, and all three compounds inhibited the aggregation of amoebae that is normally triggered by starvation. PMID- 9732447 TI - Isolation and characterization of desulfovibrio senezii sp. nov., A halotolerant sulfate reducer from a solar saltern and phylogenetic confirmation of desulfovibrio fructosovorans as a new species AB - A new halotolerant Desulfovibrio, strain CVLT (T = type strain), was isolated from a solar saltern in California. The curved, gram-negative, nonsporeforming cells (0.3 x 1.0-1.3 &mgr;m) occurred singly, in pairs, or in chains, were motile by a single polar flagellum and tolerated up to 12.5% NaCl. Strain CVLT had a generation time of 60 min when grown in lactate-yeast extract medium under optimal conditions (37 degreesC, pH 7.6, 2.5% NaCl). It used lactate, pyruvate, cysteine, or H2/CO2 + acetate as electron donors, and sulfate, sulfite, thiosulfate, or fumarate as electron acceptors. Elemental sulfur, nitrate, or oxygen were not used. Sulfite and thiosulfate were disproportionated to sulfate and sulfide. The G+C content of the DNA was 62 mol%. Phylogenetic analysis revealed that Desulfovibrio fructosovorans was the nearest relative. Strain CVLT is clearly different from other Desulfovibrio species, and is designated Desulfovibrio senezii sp. nov. (DSM 8436). PMID- 9732448 TI - Introductory statement PMID- 9732449 TI - The journal and its field: a case of co-evolution. PMID- 9732450 TI - The genetic code is one in a million. AB - Statistical and biochemical studies of the genetic code have found evidence of nonrandom patterns in the distribution of codon assignments. It has, for example, been shown that the code minimizes the effects of point mutation or mistranslation: erroneous codons are either synonymous or code for an amino acid with chemical properties very similar to those of the one that would have been present had the error not occurred. This work has suggested that the second base of codons is less efficient in this respect, by about three orders of magnitude, than the first and third bases. These results are based on the assumption that all forms of error at all bases are equally likely. We extend this work to investigate (1) the effect of weighting transition errors differently from transversion errors and (2) the effect of weighting each base differently, depending on reported mistranslation biases. We find that if the bias affects all codon positions equally, as might be expected were the code adapted to a mutational environment with transition/transversion bias, then any reasonable transition/transversion bias increases the relative efficiency of the second base by an order of magnitude. In addition, if we employ weightings to allow for biases in translation, then only 1 in every million random alternative codes generated is more efficient than the natural code. We thus conclude not only that the natural genetic code is extremely efficient at minimizing the effects of errors, but also that its structure reflects biases in these errors, as might be expected were the code the product of selection. PMID- 9732451 TI - Accelerated evolution of cytochrome b in simian primates: adaptive evolution in concert with other mitochondrial proteins? AB - We have sequenced the cytochrome b gene of Horsfield's tarsier, Tarsius bancanus, to complete a data set of sequences for this gene from representatives of each primate infraorder. These primate cytochrome b sequences were combined with those from representatives of three other mammalian orders (cat, whale, and rat) in an analysis of relative evolutionary rates. The nonsynonymous nucleotide substitution rate of the cytochrome b gene has increased approximately twofold along lineages leading to simian primates compared to that of the tarsier and other primate and nonprimate mammalian species. However, the rate of transversional substitutions at fourfold degenerate sites has remained uniform among all lineages. This increase in the evolutionary rate of cytochrome b is similar in character and magnitude to that described previously for the cytochrome c oxidase subunit II gene. We propose that the evolutionary rate increase observed for cytochrome b and cytochrome c oxidase subunit II may underlie an episode of coadaptive evolution of these two proteins in the mitochondria of simian primates. PMID- 9732452 TI - Diversifying selection governs sequence polymorphism in the major adhesin proteins fimA, papA, and sfaA of Escherichia coli. AB - Fimbriae or pili are essential adherence factors usually found in pathogenic bacteria to aid colonization of host cells. Three major structural pilin genes, fimA, sfaA, and papA, from Escherichia coli natural isolates were examined and nucleotide sequence data revealed elevated levels of both synonymous and nonsynonymous site variation at these loci. Examination of synonymous site variation shows a fivefold increase in fimA sites, relative to the housekeeping gene mdh; and similarly the sfaA and papA genes have increased synonymous sites variation relative to fimA. Nonsynonymous site variation is also elevated at all three loci but, in particular, at the papA locus (kN = 0.44). The kN/kS ratio for the three genes are among the highest yet reported for E. coli genes. Regional variation in nucleotide polymorphism within each of the genes reveal hypervariable segments where nonsynonymous substitutions exceed synonymous substitutions. We propose that at the fimA, papA, and sfaA genes, diversifying selection has brought about the increase levels of polymorphism. PMID- 9732453 TI - An evaluation of measures of synonymous codon usage bias. AB - Synonymous codons are not generally used at equal frequencies, and this trend is observed for most genes and organisms. Several methods have been proposed and used to estimate the degree of the nonrandom use of the different synonymous codons. The estimates obtained by these methods, however, show different levels of both precision and dispersion when coding regions of a finite number of codons are under analysis. Here, we present a study, based on computer simulation, of how the different methods proposed to evaluate the nonrandom use of synonymous codons are affected by the length of the coding region analyzed. The results show that some of these methods are heavily influenced by the number of codons and that the comparison of codon usage bias between coding regions of different lengths shows a methodological bias under different conditions of nonrandom use of synonymous codons. The study of the dispersion of the estimates obtained by the different methods gives, on the other hand, an indication of the methods to be applied to compare values of codon usage bias among coding regions of equivalent length. PMID- 9732454 TI - The mouse gene encoding the testis-specific isoform of Poly(A) binding protein (Pabp2) is an expressed retroposon: intimations that gene expression in spermatogenic cells facilitates the creation of new genes. AB - The gene encoding the testis-specific isoform of mouse poly(A) binding protein (Pabp2) has been isolated and sequenced. Unexpectedly, comparison of the sequence of genomic and cDNAs demonstrated that the Pabp2 gene lacks introns, whereas all other functional Pabp genes in plants, amphibians, and mammals contain introns. Thus, the mouse Pabp2 gene is a retroposon, created by synthesizing a reverse transcriptase copy of a processed mRNA and inserting the copy into the genome. The Pabp2 retroposon is unusual because it is functional: previous work demonstrates that its promoter drives the accumulation of Pabp2 mRNA in meiotic and early haploid spermatogenic cells, and the Pabp2 mRNA encodes a protein whose size and RNA-binding specificities are characteristic of PABP in plants, yeast, and mammals (Kleene et al. 1994). Two novel factors can be implicated in the retention of function of the Pabp2 retroposon. First, the promoter of the Pabp2 gene is not derived from its intron-containing progenitor, Pabp1. Second, mRNAs encoding somatic PABP isoform, PABP1, are present at high levels in meiotic and haploid spermatogenic cells. Both features contrast with the phosphoglycerate kinase 2 retroposon, which is believed to compensate for the depletion of the somatic isoform due to X-chromosome inactivation in meiotic spermatogenic cells. We also document that more functional retroposons are expressed in meiotic and haploid spermatogenic cells than in any other tissue and speculate that transcriptional derepression in spermatogenic cells favors the creation of expressed retroposons. PMID- 9732455 TI - Molecular evolution of P transposable elements in the Genus drosophila. II. The obscura species group. AB - A phylogenetic analysis of P transposable elements in the Drosophila obscura species group is described. Multiple P sequences from each of 10 species were obtained using PCR primers that flank a conserved region of exon 2 of the transposase gene. In general, the P element phylogeny is congruent with the species phylogeny, indicating that the dominant mode of transmission has been vertical, from generation to generation. One manifestation of this is the distinction of P elements from the Old World obscura and subobscura subgroups from those of the New World affinis subgroup. However, the overall distribution of elements within the obscura species group is not congruent with the phylogenetic relationships of the species themselves. There are at least four distinct subfamilies of P elements, which differ in sequence from each other by as much as 34%, and some individual species carry sequences belonging to different subfamilies. P sequences from D. bifasciata are particularly interesting. These sequences belong to two subfamilies and both are distinct from all other P elements identified in this survey. Several mechanisms are postulated to be involved in determining phylogenetic relationships among P elements in the obscura group. In addition to vertical transmission, these include retention of ancestral polymorphisms and horizontal transfer by an unknown mating-independent mechanism. PMID- 9732456 TI - Full-length L1 elements have arisen recently in the same 1-kb region of the gorilla and human genomes. AB - New copies of the mammalian retrotransposon L1 arise in the germline at an undetermined rate. Each new L1 copy appears at a specific evolutionary time point that can be estimated by phylogenetic analysis. In humans, the active L1 sequence L1.2 resides at the genomic locus LRE1. Here we analyzed the region surrounding the LRE1 locus in humans and gorillas to determine the evolutionary history of the region and to estimate the age of L1.2. We found that the region was composed of an ancient L1, L1Hs-Lrg, which was significantly divergent from all other L1 sequences available in the databases. We also determined that L1.2 was absent from the gorilla genome and arose in humans after the divergence of gorilla and human lineages. In the gorilla LRE1 region, we discovered a different full-length L1 element, L1Gg-1, which was allelic and present at a high gene frequency in gorillas but absent from other primates. We determined the nucleotide sequence of L1Gg-1 and found that it was 98% identical to L1.2, suggesting a close relationship between active L1s in gorillas and humans. PMID- 9732457 TI - The complete mitochondrial DNA sequence of the pig (Sus scrofa). AB - The complete mitochondrial genome sequence of the pig, Sus scrofa, was determined. The length of the sequence presented is 16,679 nucleotides. This figure is not absolute, however, due to pronounced heteroplasmy caused by variable numbers of the motif GTACACGTGC in the control region of different molecules. A phylogenetic study was performed on the concatenated amino acid and nucleotide sequences of 12 protein-coding genes of the mitochondrial genome. The analysis identified the pig (Suiformes) as a sister group of a cow/whale clade, making Artiodactyla paraphyletic. The split between pig and cow/whale was molecularly dated at 65 million years before present. PMID- 9732458 TI - Conflict among individual mitochondrial proteins in resolving the phylogeny of eutherian orders. AB - The phylogenetic relationship among primates, ferungulates (artiodactyls + cetaceans + perissodactyls + carnivores), and rodents was examined using proteins encoded by the H strand of mtDNA, with marsupials and monotremes as the outgroup. Trees estimated from individual proteins were compared in detail with the tree estimated from all 12 proteins (either concatenated or summing up log-likelihood scores for each gene). Although the overall evidence strongly suggests ((primates, ferungulates), rodents), the ND1 data clearly support another tree, ((primates, rodents), ferungulates). To clarify whether this contradiction is due to (1) a stochastic (sampling) error; (2) minor model-based errors (e.g., ignoring site rate variability), or (3) convergent and parallel evolution (specifically between either primates and rodents or ferungulates and the outgroup), the ND1 genes from many additional species of primates, rodents, other eutherian orders, and the outgroup (marsupials + monotremes) were sequenced. The phylogenetic analyses were extensive and aimed to eliminate the following artifacts as possible causes of the aberrant result: base composition biases, unequal site substitution rates, or the cumulative effects of both. Neither more sophisticated evolutionary analyses nor the addition of species changed the previous conclusion. That is, the statistical support for grouping rodents and primates to the exclusion of all other taxa fluctuates upward or downward in quite a tight range centered near 95% confidence. These results and a site-by site examination of the sequences clearly suggest that convergent or parallel evolution has occurred in ND1 between primates and rodents and/or between ferungulates and the outgroup. While the primate/rodent grouping is strange, ND1 also throws some interesting light on the relationships of some eutherian orders, marsupials, and montremes. In these parts of the tree, ND1 shows no apparent tendency for unexplained convergences. PMID- 9732459 TI - Identification of major phylogenetic branches of inhibitory ligand-gated channel receptors. AB - The gene superfamily of ligand-gated ion channel (LGIC) receptors is composed of members of excitatory LGIC receptors (ELGIC) and inhibitory LGIC receptors (ILGIC), all using amino acids as ligands. The ILGICs, including GABAA, Gly, and GluCl receptors, conduct Cl- when the ligand is bound. To evaluate the phylogenetic relationships among ILGIC members, 90 protein sequences were analyzed by both maximum-parsimony and distance matrix-based methods. The strength of the resulting phylogenetic trees was evaluated by means of bootstrap. Four major phylogenetic branches are recognized. Branch I, called BZ, for the majority of the members are known to be related to benzodiazepine binding, is subdivided into IA, composed of all GABAA receptor alpha subunits, and IB, composed of the gamma and epsilon subunits, which are shown to be tightly linked. Branch II, named NB for non-benzodiazepine binding, and consisting of GABAA receptor beta, delta, pi, and rho subunits, is further subdivided into IIA, containing beta subunits; IIB, containing delta, and pi subunits; and IIC, containing rho subunits. Branch IIIA, composed of vertebrate Gly receptors, is loosely clustered with Branch IIIB, composed of invertebrate GluCl receptors, to form Branch III, which is designated NA for being non-GABA responsive. Branch IV is called UD for being undefined in specificity. The existence of primitive forms of GABAA receptor non-beta subunits in invertebrates is first suggested by the present analysis, and the identities of sequences p25123 from Drosophila melanogaster, s34469 from Lymnaea stagnalis, and u14635 and p41849 from C. aenorhabditis elegans are determined to be different from their previously given annotations. The proposed branching classification of ILGICs provides a phylogenetic map, based on protein sequences, for tracing the evolutionary pathways of ILGIC receptor subunits and determining the identities of newly discovered subunits on the basis of their protein sequences. PMID- 9732460 TI - Evolution of the primate androgen receptor: a structural basis for disease. AB - Androgen effects mediated by the androgen receptor (AR) are essential for male reproductive development and virilization. Comparison of AR DNA coding sequence from five primate species, Homo sapiens (human), Pan troglodytes (chimpanzee), Papio hamadryas (baboon), Macaca fascicularis (macaque), and Eulemur fulvus collaris (collared brown lemur), supports their phylogeny with complete conservation of the DNA and steroid binding domain protein sequence. A linear increase in trinucleotide repeat expansion of homologous CAG and GGC sequences occurs in the NH2-terminal transcriptional activation region and is proportional to the time of species divergence. A serine phosphate/glutamine repeat interaction is observed where increasing CAG repeat length is associated with an increased rate of serine 94 phosphorylation. Disparity in the calculated and apparent molecular weight with CAG repeat expansion of an AR NH2-terminal fragment suggests self-aggregation with increasing glutamine repeat length into the pathological range. These results suggest that a CAG/glutamine repeat expanded during divergence of the higher primate species, which may have a direct effect on AR structure and support a common pathway in CAG trigenic diseases in the pathophysiology of neurodegeneration observed in X-linked spinal bulbar and muscular atrophy. PMID- 9732461 TI - Identification and expression of the SOS response, aidB-like, gene in the marine sponge Geodia cydonium: implication for the phylogenetic relationships of metazoan acyl-CoA dehydrogenases and acyl-CoA oxidases. AB - Sponges (Porifera) are the phylogenetically oldest metazoan organisms. From one member of the siliceous sponges, Geodia cydonium, the cDNA encoding a putative SOS protein, the AidB-like protein of the Ada system from bacteria, was isolated and characterized. The cDNA, GCaidB, comprises an open reading frame of 446 amino acid (aa) residues encoding a polypeptide with a calculated Mr of 49,335. This molecule shows high similarity to the bacterial AidB proteins from Mycobacterium tuberculosis and Escherichia coli and somewhat lower similarities to acyl-CoA dehydrogenases (ADHs) and acyl-CoA oxidases (AOXs). Northern blot analysis confirmed the presence of the complete transcript. The deduced sponge aa sequence, GC_aidB, possesses the two characteristic acyl-CoA dehydrogenase signatures 1 and 2. Incubation of the sponge with N-methyl-N'-nitro-N nitrosoguanidine causes a strong increase in the 2.1-kb large transcript of GCaidB; maximal expression is seen after 24 h of incubation with this DNA methylating agent. ADHs and AOXs can be grouped, depending on the position of the catalytically important Glu residue, into the Glu-Gly (Glu adjacent to Gly) class and the Glu-Arg (Glu adjacent to Arg) class. The phylogenetically oldest metazoan AidB-like molecule, GC_aidB of G. cydonium, belongs to the Glu-Gly class of ADHs. Phylogenetic analyses of the Glu-Gly class enzymes, with the described AidB-like protein from G. cydonium and the bacterial AidB polypeptides, together with metazoan ADHs and AOXs, revealed that the AidB(-like) proteins diverged first from a common ancestor, while the eukaryotic AOX and ADA polypeptides as well as the GHDs appeared later. According to the analyses, the very long-chain ADHs are older than the medium-chain, short-chain, and branched-chain ADHs. Inclusion of the phylogenetical oldest member of the Glu-Arg class of enzymes, the bacterial ADH-CaiA sequence in these analyses, revealed that this class of enzymes appeared later in evolution and arose from the Glu-Gly class perhaps after gene duplication. PMID- 9732464 TI - Early metazoan divergence was about 830 million years ago. AB - From a total of 22 nuclear genes, we estimate that the divergence time between Drosophila and vertebrates was about 830 million years ago (mya), which is significantly (1% level) earlier than the Cambrian explosion indicated by the early triploblastic fossils (<600 mya). PMID- 9732462 TI - The origin of trypsin: evidence for multiple gene duplications in trypsins. AB - The trypsin family of serine proteases is one of the most studied protein families, with a wealth of amino acid sequence information available in public databases. Since trypsin-like enzymes are widely distributed in living organisms in nature, likely evolutionary scenarios have been proposed. A novel methodology for Fourier transformation of biological sequences (FOTOBIS) is presented. The methodology is well suited for the identification of the size and extent of short repeats in protein sequences. In the present paper the trypsin family of enzymes is analyzed with FOTOBIS and strong evidence for tandem gene duplication is found. A likely evolutionary path for the development of present-day trypsins involved an intrinsic extensive tandem gene duplication of a small DNA fragment of 15-18 nucleotides, corresponding to five or six amino acids. This ancestral trypsin gene was subsequently duplicated, leading to the earliest version of a full-sized trypsin, from which the contemporary trypsins have developed. PMID- 9732463 TI - Heterologous gene expression in an Escherichia coli population under starvation stress conditions. AB - A novel system to study the evolution of transcription signals in heterologous systems under selective starvation conditions is described. It is based on the plasmid-mediated transfer of his biosynthetic genes from Azospirillum brasilense into a heterologous Escherichia coli mutant population lacking histidine biosynthetic ability. We show that under highly selective stressful conditions, genetic changes in the donor plasmid lead to mutated sequences that are efficiently recognized as promoters by the E. coli RNA polymerase. PMID- 9732465 TI - Human cell line toxicity of binary and ternary chemical mixtures in comparison to individual toxic effects of their components. AB - Contaminated site projects involve health risk assessment procedures that must consider potential interactive effects of present contaminants. In order to establish a quick and reliable method that accounts for smaller than additive, additive, or greater than additive toxic effects, this study compared the cytotoxicity of 34 binary and ternary chemical mixtures of four structurally different chemicals to HeLa cells. Further, five blind samples of these mixtures or their components were tested to determine the ability to identify unknown mixtures. The colorimetric MTS in vitro cytotoxicity assay was used to detect cytotoxic effects after the cells were exposed for 1 h to serial dilutions of the mixtures. Experimental cytotoxicity data were compared and set against data predicted by a mathematical algorithm. They mainly showed additive effects of the components in mixture but also identified smaller than additive and greater than additive effects. A subjective classification scheme allowed evaluation of the toxicity of the blind samples in comparison to results from the study on binary and ternary mixtures tested before. This scheme focused on quantitative cytotoxicity data as well as on the slope of the concentration-effect curves and demonstrated the use of the MTS assay for human health risk assessments in the context of contaminated sites. PMID- 9732466 TI - Spatial and temporal trends of paraquat, diquat, and difenzoquat contamination in water from marsh areas of the valencian community (Spain) AB - The levels and distribution of diquat, paraquat, and difenzoquat were determined by solid phase extraction (SPE) and high-performance liquid chromatography (HPLC) in water samples from irrigation channels, rivers, and lagoons taken during 1 year from three different marsh areas of the Valencian community. These areas are representative of the typical Mediterranean coastal ecosystems. All three compounds were detected. Diquat was found most frequently at all the sampling sites. Although the spatial distribution of diquat and paraquat showed a maximum concentration near the fields where they were originally applied, their location fluctuated due to irregular large spill and/or loading. The herbicide concentration tended to be highest during the summer (June, July, and August) because these are the months with the least rainfall and highest evaporation rates, when weeds grow best and pesticides are needed more often. The average concentration found for diquat was 0.09 &mgr;g/L, with a maximum of 3.10 &mgr;g/L. The average concentration for paraquat was 0.01 &mgr;g/L, with a maximum of 3.95 &mgr;g/L. Samples that were below the method detection limit are included in the mean calculation as zero. Difenzoquat was only detected in one sample at a concentration of 1.75 &mgr;g/L. PMID- 9732467 TI - The environmental occurrence of herbicides: the importance of degradates in ground water. AB - Numerous studies are being conducted to investigate the occurrence, fate, and effects on human health and the environment from the extensive worldwide use of herbicides to control weeds. Few studies, however, are considering the degradates of these herbicides in their investigations. Our study of herbicides in aquifers across Iowa found herbicide degradates to be prevalent in ground water, being detected in about 75% of the wells sampled. With the exception of atrazine, the frequencies of detection in ground water for a given herbicide increased multifold when its degradates were considered. Furthermore, a majority of the measured concentration for a given herbicide was in the form of its degradates even for a relatively persistent compound such as atrazine. For this study, degradates comprised from 60 to over 99% of a herbicide's measured concentration. Because herbicide degradates can have similar acute and chronic toxicity as their parent compounds, these compounds have environmental significance as well as providing a more complete understanding of the fate and transport of a given herbicide. Thus, it is essential that degradates are included in any type of herbicide investigation. PMID- 9732469 TI - Photolysis of phloxine B in water and aqueous solutions AB - Phloxine B (2',4',5',7'-tetrabromo-4,5,6,7-tetrachlorofluorescein disodium salt) as a potential photoactive insecticide was rapidly photodegraded in water under various light sources. Two major photolytic products characterized were 2',4',5' tribromo-4,5,6, 7-tetrachlorofluorescein and 4',5'-dibromo-4,5,6, 7 tetrachlorofluorescein. The photolysis rates of phloxine B were influenced by various factors including salts in medium, sample pH, and light sources. Half lives (t(1/2)) of phloxine B spiked in different water samples and 2% NaCl solution at 29 +/- 1 degreesC ranged from 0.70 to 1.28, 26.3 to 115, and 14.1 to 46.2 hours under 254 nm, 365 nm, and cool white fluorescent lights, respectively. Half-lives of phloxine B in tap, stream, or seawater in a beaker were from 10 to 13 min under sunlight at ambient air temperature. In a range of buffer pH 6-8 at 29 +/- 1 degreesC, phloxine B photodegraded slightly faster in acidic solution than in basic solution. The photolysis t(1/2) of phloxine B at 29 +/- 1 degreesC was 25, 32, 128, and 755 min in the buffered NaF, NaCl, NaBr, and NaI solutions, respectively. The t(1/2) of phloxine B was 31 min when phloxine B was dissolved in the sodium phosphate buffer as control. Sodium iodide and ammonium iodide photostabilized phloxine B 24 and 27 folds, respectively, when it was compared with the buffer control. PMID- 9732468 TI - Organophosphate and carbamate insecticides in agricultural waters and cholinesterase (ChE) inhibition in common carp (Cyprinus carpio). AB - Cholinesterase (ChE) activity was used as a biomarker for assessing exposure of common carp (Cyprinus carpio) to organophosphate and carbamate insecticides from irrigated agricultural waters. Carp were collected from a lake (Royal Lake) that receives most of its water from irrigation return flows and from a reference lake (Billy Clapp Lake) outside of the irrigation system. Results indicated that the mean whole-brain ChE activity of carp from Royal Lake (3.47 micromol/min/g tissue) was 34.2% less than that of carp from Billy Clapp Lake (5.27 micromol/min/g tissue) (p = 0.003). The depressed ChE activity in brain tissue of Royal Lake carp was in response to ChE-inhibiting insecticides detected in water samples in the weeks prior to tissue sampling; the most frequently detected insecticides included chlorpyrifos, azinphos-methyl, carbaryl, and ethoprop. Neither sex nor size appears to be a covariable in the analysis; ChE activity was not correlated with fish length or weight in either lake and there was no significant difference in ChE activity between the two sexes within each lake. Although organophosphate and carbamate insecticides can break down rapidly in the environment, this study suggests that in agricultural regions where insecticides are applied for extended periods of the year, nontarget aquatic biota may be exposed to high levels of ChE-inhibiting insecticides for a period of several months. PMID- 9732470 TI - Influence of trophic status on the toxic effects of a herbicide: A microcosm study AB - Naturally derived microbial communities were developed in the laboratory under three nutrient regimes by manipulating phosphate and nitrate concentrations. Resulting communities differed in both functional and structural attributes. Low nutrient microcosms (0.05 mg N-NO3-/L + 0.01 mg P-PO4-3/L) showed the sharpest differences. Medium (0.5 mg N-NO3-/L + 0.1 mg P-PO4-3/L) and high (5.0 mg N-NO3 /L + 1.0 mg P-PO4-3/L) nutrient treatments differed in total algal biomass and algal community composition. After a 25-day acclimation period, a single dose of the herbicide diquat (3.5 mg/L) was added to test the response of the microbial communities to herbicide stress. Regardless of nutrient regime, diquat-dosed microcosms had decreased electron transport system activity (ETSA), an almost complete absence of cyanobacteria, and reduced gross photosynthesis (GP), respiration, and pH relative to undosed microcosms. Inorganic nutrients (PO4-3, NO3-) were released from the stressed algal communities, probably as a result of their altered metabolism. Alkaline phosphatase activity (APA), total microbial biomass (estimated as protein), algal biomass (estimated as chlorophyll), and relative abundance of green algal taxa proved highly insensitive to herbicide action. Nutrient treatments had a small influence on toxicant effects; the magnitude of the herbicide effects was comparable across nutrient levels. Only the capacity of recovery from the toxic stress was affected by trophic status. At the end of the study period, ETSA had recovered to control values in high nutrient microcosms but not in medium and low ones. Microcosm pH, and to a lesser extent GP, showed recovery under both high and medium nutrient treatments. Trophic status affected the diquat disappearance rate; the herbicide persisted longer in low nutrient microcosms than in high and medium nutrient ones. Differences in recovery capacity may stem from higher nutrient level microcosms reaching less toxic herbicide levels in a shorter period of time. PMID- 9732471 TI - Characterization of organotin-resistant bacteria from boston harbor sediments AB - Organotins are widely used in agriculture and industry. They are toxic to a variety of organisms including bacteria, although little is known of their physiology and ecology. Bacteria resistant to six organotins-tributyltin (TBT), dibutyltin (DBT), monobutyltin (MBT), triphenyltin (TPT), diphenyltin (DPT), and monophenyltin (MPT)-were isolated from Boston Harbor sediments, Massachusetts, USA. Bacteria resistant to each of the organotins, except DPT, were isolated directly from estuarine sediments. Viability of the organotin-resistant bacteria on serial transfer in the laboratory ranged from 80 to 91%. Each isolate was screened for resistance to the other organotins. All of 250 isolates were resistant to at least two organotins. No DPT-resistant isolates were found on initial isolation on DPT, although there was DPT resistance among the other organotin-resistant bacteria. Eighty percent of TBT-resistant bacteria were TPT resistant, suggesting that antifouling paints containing TPT will not be a suitable substitute for TBT in paints designed to inhibit microbial biofilms. Debutylation reduced toxicity in some cases while dephenylation did not. Thus, even though trisubstituted organotins are generally believed to be more toxic than di- or monosubstituted organotins, this may not always be the case, and more than one mechanism of resistance may be involved. All the bacteria were resistant to at least six of eight heavy metals tested, suggesting that resistance to heavy metals may be associated with resistance to organotins. PMID- 9732472 TI - Sensitivity to copper in a ciliate as a possible component of biological monitoring in the lagoon of venice AB - The impact of copper was studied in cultures of the microbenthic organism Euplotes vannus (Ciliophora, Hypotrichida). This ciliate was isolated from sediment collected from a particular area in the Lagoon of Venice, which may be considered almost unpolluted by heavy metals. The effects of copper exposure in the laboratory on growth, metal accumulation, total acid-soluble thiol levels, and glutathione levels were examined, together with morphological alterations. E. vannus exhibited tolerance toward copper up to a concentration exposure of 0.2 &mgr;g Cu/ml, at which cell growth rate overlapped that of controls. Copper accumulated up to 239 &mgr;g/g dry wt after exposure to 0.4 &mgr;g Cu/ml, and morphological alterations were evident in cells exposed to concentrations from this value upward. Processes of vegetative reorganization involving nuclear apparatus, membranelles, cirri, and cortex could be observed after 4 days of exposure to 0.4 &mgr;g Cu/ml. Laboratory experiments with cultures of ciliates of cosmopolitan distribution such as Euplotes species in controlled conditions indicated the value of these organisms, which constitute a simple model for a monitoring system suitable for prediction in multicellular organisms. PMID- 9732473 TI - The toxicity of margosan-O, a product of neem seeds, to selected target and nontarget aquatic invertebrates AB - Margosan-O, an insecticide formulated from extracts of neem tree (Azadirachta indica) seed kernels, besides being toxic, also has feeding, oviposition deterring, and growth-inhibitory effects on insects. This product, registered in the United States for ornamental plants, has been proposed for food crop use. However, little information exists on its effects on aquatic organisms. This study investigated toxicity of Margosan-O to the mosquito Culex spp., a possible target species, and to nontarget species-two crustaceans, Daphnia magna, Hyalella azteca, and a dipteran, Chironomus riparius. The 48-h EC50 value of 105 mg L-1 for Culex spp. was significantly more toxic than for C. riparius (281 mg L-1), not significantly different from D. magna (125 mg L-1) but was significantly less toxic than for H. azteca (71 mg L-1). A concentration of 20-30 mg L-1 caused growth inhibitory effects in Culex spp. and C. riparius larvae and 40 and 84 mg L 1 affected growth and reproduction in H. azteca and D. magna, respectively. Margosan-O may not be suitable for mosquito control since the concentrations required to control emergence may have some nontarget effects. Alternatively, the agricultural application of Margosan-O is also not expected to reduce the survival or produce growth and reproductive effects in nontarget aquatic organisms. However, based on estimated concentrations of less than 10 mg L-1 in adjacent shallow bodies of water and recommendations for repeated applications, there should be concern that the threshold for chronic toxicity is too narrow. PMID- 9732474 TI - The acute toxicity of lindane to hyalella azteca and the development of a sublethal bioassay based on precopulatory guarding behavior AB - Acute and sublethal toxicity of the organochlorine insecticide lindane to the amphipod crustacean Hyalella azteca was investigated. Acute experiments were conducted for a maximum test exposure period of 240 h with adult and neonate H. azteca. Median lethal concentrations (LC50s) determined for adult Hyalella included a 48-h LC50 of 47.6 &mgr;g/L and 240-h LC50 of 26.9 &mgr;g/L. For neonate H. azteca 24-, 48-, and 240-h LC50s were 29.5, 14.8, and 9.8 &mgr;g lindane/L, respectively. Neonate H. azteca were approximately three times more sensitive than adults. Two sublethal toxicity bioassays were developed based on the direct and indirect disruption of the precopulatory or mate guarding behavior of Hyalella. This reproductive behavior is readily quantifiable and of ecological significance as it is a vital component of the mating success of the species. The direct disruption bioassay examined the separation of precopulatory pairs maintained in control water and a range of lindane concentrations during a 24-h exposure period. Median separation times (ST50s) were determined and the LOEC was 24.4 &mgr;g lindane/L. The indirect disruption bioassay consisted of a test exposure period of just 4 h after which an invertebrate anesthetic solution was administered to induce separation of precopulatory pairs. The LOEC was 17.3 &mgr;g lindane/L, suggesting that the indirect precopulatory separation bioassay was comparable to the 24-h direct separation study. Both bioassays are rapid, relatively simple to perform, and have yielded effect concentrations that correspond with LC50 values determined using adult and neonate H. azteca life stages over more prolonged lindane exposures. Following some modification, these behavioral bioassays may be suitable for use in the hazard evaluation of sediments and for deployment as in situ toxicity tests. PMID- 9732475 TI - Chronic effects of the herbicide diuron on freshwater cladocerans, amphipods, midges, minnows, worms, and snails. AB - The chronic effects of the herbicide diuron on survival and reproduction of Daphnia pulex, and survival and growth of the amphipod Hyalella azteca, the midge Chironomus tentans, juvenile and embryo/larval fathead minnows, Pimephales promelas, annelid worms, Lumbriculus variegatus, and snails, Physa gyrina, were determined in laboratory static and static-renewal tests. D. pulex 96-h and 7-day LC50 values were 17.9 and 7.1 mg/L; 7-day LOAEL and NOAEL values based on mortality and reproduction were 7.7 and 4.0 mg/L. H. azteca 96-h and 10-day LC50 values were 19.4 and 18.4 mg/L; 10-day LOAEL and NOAEL values based on survival and reduced weight were 15.7 and 7.9 mg/L. C. tentans 10-day LC50 value was 3.3 mg/L; 10-day LOAEL and NOAEL values based on growth were 7.1 and 3.4 mg/L, and 3.4 and 1.9 mg/L based on mortality. Juvenile fathead minnows had a 10-day LC50 of 27.1 mg/L and 10-day LOAEL and NOAEL values based on growth of 3.4 and <3.4 mg/L. The fathead minnow embryo-larval test had a 7-day LC50 value of 11.7 mg/L and 7-day LOAEL and NOAEL values based on reduced growth of 8.3 and 4.2 mg/L. L. variegatus had 10-day LOAEL and NOAEL values based on reduced weight of 3.5 and 1.8 mg/L. P. gyrina had 10-day LOAEL and NOAEL values based on reduced weight of 22.8 and 13.4 mg/L. Laboratory effects concentrations were higher that those found in normal field application situations, except in areas of localized pooling after recent herbicide applications, indicating that there would probably be little harm to these fish and invertebrates from diuron exposure in the field. PMID- 9732477 TI - Heavy metals inhibit limb regeneration in horseshoe crab larvae AB - We studied the effects of heavy metals on the regeneration of walking legs in horseshoe crabs (Limulus polyphemus). The second walking leg was amputated in embryos (stage 20 and 21) and first instar (trilobite) larvae, and the length and morphology of the regenerated appendage was observed after molting to the second instar stage. Regeneration following continuous exposure to TBT (0. 001-100 mg/L), mercury (0.001-100 mg/L), cadmium (0.01-100 mg/L), chromium (0.1-100 mg/L), lead (0.1-100 mg/L), and copper (1-100 mg/L) was measured relative to regeneration in seawater. Although regeneration was incomplete in controls, treatment with heavy metals led to smaller and/or malformed legs. The impacts of heavy metals on survival, molting, and regeneration of horseshoe crab larvae were ranked as follows: organotin > Hg > Cd > Cr > Zn > Pb >== Cu. Cu and Pb did not inhibit regeneration, even at 100 mg/L. TBT, Hg, Cd, Cr, and Zn inhibited the regeneration of appendages, although first instar larvae successfully molted into second instars even after treatment. Regeneration was comparable to seawater controls in less than 2.5 mg/L Zn. In 5.0 and 10.0 mg/L Zn, regeneration was inhibited and the length of regenerated appendages remained shorter in all second instars. Larvae treated with 10 mg/L Zn for 1-week intervals during the molt cycle showed similar patterns of regeneration. The regeneration of claws was not all or none, and formation of the claw was proportional to the length of regenerated appendages. Limb regeneration in horseshoe crab larvae may be a useful model system for the study of pollutant impacts. PMID- 9732476 TI - Heavy metals alter the survival, growth, metamorphosis, and antipredatory behavior of Columbia spotted frog (Rana luteiventris) tadpoles. AB - Amphibian populations appear to be declining around the world. Although there is no single cause, one factor may be pollution from heavy metals. As a result of mining in the Silver Valley of Idaho, heavy metals have been released into habitats containing many species of sensitive organisms, including spotted frogs (Rana luteiventris). While the gross extent of pollution has been well documented, the more subtle behavioral effects of heavy metals such as lead, zinc, and cadmium are less well studied. We tested the effects of heavy metals on the short-term survival (LC50) of spotted frog tadpoles. Compared to single metals, metals presented together were toxic at lower doses. We also raised the tadpoles in outdoor mini-ecosystems containing either a single heavy metal or soil from an EPA Superfund site in the Silver Valley known to be composed of numerous heavy metals. Exposure to Silver Valley soil resulted in delayed metamorphosis. We tested the ability of metal-exposed tadpoles to detect and respond to chemical cues emanating from predacious rainbow trout. We found that high levels of Silver Valley soil, medium levels of zinc, and medium and high levels of lead resulted in a decreased fright response. Low levels of cadmium, zinc, and lead did not cause a significant effect, but low levels of soil did result in a decreased fright response. Heavy metals may alter interactions between tadpoles and their predators. PMID- 9732478 TI - Altered growth and metabolism of an estuarine shrimp (Palaemonetes pugio) during and after metamorphosis onto fenvalerate-laden sediment. AB - Dry weight (W), carbon (C), nitrogen (N), and energy (E) (calculated) accumulation were measured in the estuarine grass shrimp, Palaemonetes pugio, throughout larval development and during the first 2 weeks as postlarvae in seawater over sediment containing the pyrethroid insecticide fenvalerate (SCF; nominal concentrations of 1, 10, and 100 microgram fenvalerate kg-1 sediment). The influence of fenvalerate-laden sediment on shrimp growth and utilization patterns of C, N, and E was dependent on fenvalerate concentration, age of shrimp, and whether shrimp were premetamorphic or postmetamorphic in development. The fenvalerate concentration in the sediment, which ultimately inhibited larval metamorphosis (100 microgram fenvalerate kg-1 sediment), significantly reduced W accumulation in developing larvae and in postlarvae growing on the sediment for an equivalent time. Accumulation of C, N, and E varied not only with concentration of SCF, but differed between pelagic larvae developing in water above SCF and newly settled postlarvae growing in direct contact with SCF. Larvae developing above >/=10 microgram kg-1 SCF contained significantly less N, while postlarval shrimp settling onto >/=10 microgram kg-1 SCF accumulated significantly less C and E. Measurable variations in growth and energy reserves of toxicant-sensitive life stages in response to environmentally realistic insecticide exposures have a direct link to ecological consequences of toxic stress and may be useful as biomarkers to diagnose early damage in estuarine populations. PMID- 9732479 TI - A study of the lethal and sublethal toxicity of polyphase P-100, an antisapstain fungicide containing 3-iodo-2-propynyl butyl carbamate (IPBC), on fish and aquatic invertebrates. AB - The acute toxicity of Polyphase P-100, an antisapstain wood preservative that contains 97% 3-iodo-2-propynyl butyl carbamate (IPBC), was determined for three species of fish (coho salmon, rainbow trout, and starry flounder) and three species of aquatic invertebrates (Daphnia magna, Hyalella azteca, and Neomysis mercedis). The 96-h LC50 values for the various fish species exposed to Polyphase P-100 ranged from 95 ppb for coho smolts (Oncorhynchus kisutch) to 370 ppm for juvenile starry flounder (Platichthys stellatus). The sensitivity of coho to Polyphase P-100 was altered by their developmental stage. Coho embryos were six to nine times more tolerant of Polyphase P-100 than coho alevins, which were twice as tolerant as coho smolts. The 48-h LC50 values for the invertebrates D. magna, H. azteca, and N. mercedis were 40 ppb, 500 ppb, and 2,920 ppb, respectively. In addition to a wider range of sensitivity to Polyphase P-100 compared with the fish species, the invertebrate species were characterized by a shallower concentration-response. In acute, 24-h sublethal tests with juvenile starry flounder and rainbow trout, there was no primary or secondary stress response (changes in hematocrit, leucocrit, hemoglobin concentration, plasma lactate concentration, and plasma cortisol concentration) at concentrations up to 50% of the 96-h LC50 value. The acute toxicity of a 1:8 mixture of Polyphase P 100 and Bardac 2280 (another antisapstain compound that contains didecyldimethylammonium chloride [DDAC] as the active ingredient) was close to additive for fish, but not for invertebrate species. The acute toxicity of the mixture was seven to eight times more than additive for H. azteca, but two to three times less than additive for D. magna. Some sublethal stress responses were revealed with the mixture that were not observed with the test chemicals alone. PMID- 9732480 TI - Acute 2,4-D poisoning in tench (Tinca tinca L.): lesions in the hematopoietic portion of the kidney. AB - An experimental model was designed to study the acute lesions caused by a continuous exposure to 2,4-dichlorophenoxyacetic acid (2,4-D) disolved in water (400 mg/L) in hematopoietic kidney tissue in tench (Tinca tinca L). Fifty fish were used in this study, 15 for calculating LC50 and 35 were euthanized 1, 2, 5, 8, and 12 days postpoisoning (five treated and two controls each time). Tissue samples, fixed in 5% glutaraldehyde in 0.1 M phosphate buffer (pH 7. 2) for histopathological examination, revealed marked alteration of hematopoietic tissue, characterized by progressive swelling and cell necrosis, activation of the phagocyte system, and subsequent formation of myelin figures. Variations recorded in hematocrit and hemoglobin levels in blood samples indicated changes in membrane permeability, complementing the findings on hematopoietic tissue. The lethal dose (LC50) at 96 h demonstrated the importance of the species and chemical form used as factors in calculating a product's toxicity. PMID- 9732481 TI - Behavioral responses to atrazine and diuron in goldfish. AB - Experiments were performed in goldfish to determine the effects of a short-term exposure (24 h) to atrazine or diuron (0.5, 5, 50 microgram/L) on some behavior endpoints related to swimming and social activities. Observations were also made to assess the influence of such exposure on the behavioral responses of fish to the flow of a crude skin extract solution from conspecifics, active in social chemocommunication and producing alarm behaviors. Additive tests were run to check the behavioral responses of previously unexposed goldfish to the flow of a solution of atrazine- or diuron-contaminated water, at three concentrations (0.1, 1, 10 mg/L). Significant burst swimming reactions appeared in response to a 24-h exposure to atrazine, at the lowest concentration tested (0.5 microgram/L). A 24 h exposure to 5 microgram/L atrazine or diuron was found to induce various significant behavioral alterations in fish. At this concentration, both herbicides decreased grouping behavior and atrazine also increased surfacing activity. Herbicide-exposed fish showed a decreased grouping behavior during the flow of the skin extract solution. Sheltering was also decreased during the flow of the biological solution in fish exposed to atrazine. Moreover, fish exposed to diuron clearly displayed attraction responses to the flow of the skin solution. Previously unexposed fish showed a significant increase in burst swimming reactions in response to the flow of a solution of atrazine- or diuron contaminated water, at all concentrations tested (0.1, 1, 10 mg/L). Furthermore, the diuron-contaminated flow was found to be significantly attractive at the highest concentration. These results indicate that a short-term exposure to a relatively low concentration (5 microgram/L) of atrazine or diuron can affect various behaviors of fish not only directly but also indirectly by altering the chemical perception of natural substances of eco-ethological importance. In consideration of the basic role of olfaction in fish behavior, these results also emphasize the need for further developments on the possible effects of aquatic toxicants on olfactory-mediated behaviors. PMID- 9732482 TI - Contaminants in eggs of colonial waterbirds and hepatic cytochrome P450 enzyme levels in pipped tern embryos, Washington State. AB - Eggs of Forster's terns (Sterna forsteri) collected in 1991 from nesting colonies on Crescent Island (Columbia River) and the Potholes Reservoir in south central Washington generally contained low residues of organochlorine pesticides and metabolites, 2,3,7, 8-tetrachlorodibenzo-p-dioxin, 2,3,7,8 tetrachlorodibenzofuran, and polychlorinated biphenyls (PCBs). Hepatic cytochrome P450 enzyme activity in pipped embryos of Forster's terns from the two colonies seemed unaffected by contaminants. At Crescent Island, examination of 23 Forster's tern eggs with large embryos (19 viable [10 pipped] and four dead [two pipped]) revealed developmental abnormalities in two viable pipped embryos (missing maxilla and deformed pelvic girdle) and a viable prepipping embryo (shortened beak). Our limited sample sizes and number of compounds analyzed preclude us from determining whether or not the abnormalities are related to contaminants. No abnormalities were noted in 10 pipped eggs (nine viable and one dead at collection) of Forster's terns collected from the Potholes Reservoir colony. Eggs of Caspian terns (Sterna caspia) collected from Crescent Island in 1991 also contained generally low residues of contaminants, only one developmental abnormality was noted, and limited data indicated that cytochrome P450 enzyme activity apparently was unaffected by contaminants. Organochlorine contaminants were generally low in addled eggs of American white pelicans (Pelecanus erythrorhynchos) collected from Crescent Island in 1994. PMID- 9732483 TI - Subchronic dietary toxicity of strychnine: bobwhite quail are less sensitive than mallard ducks. AB - Separate, 28-day, subchronic studies of strychnine dietary toxicity were conducted using northern bobwhite quail (Colinus virginianus) and mallard ducks (Anas platyrhynchos). Five groups (five males five females/group) of 29-week-old quail were fed Purina(R) Game Bird Breeder Layena(R) diets containing mean (+/ SD) 484.2 (+/-17.0), 972. 6 (+/-54.0), 1,870.8 (+/-176.1), 3,516.7 (+/-68.0), and 6,083.3 (+/-269.6) microgram/g strychnine; whereas five groups of 27-week-old mallards (five males five females/group) were fed similar diets containing mean (+/-SD) 18.8 (+/-1.3), 91.1 (+/-27.3), 235.0 (+/-33. 8), 484.2 (+/-17.0), and 972.6 (+/-54.0) microgram/g strychnine. Separate "vehicle control" (0.0 microgram/g strychnine) groups (five males, five females/group) were included in each study. Strychnine toxicity was much less pronounced in quail; no observed effect concentrations (NOECs) were 972.6 (+/-54.0) and 91.1 (+/-27.3) microgram/g strychnine for quail and ducks, respectively. Several possible explanations for the species effects are offered, and some practical issues affecting the conduct of long-term, dietary toxicity studies are discussed. PMID- 9732484 TI - Retrospective study of the diagnostic criteria in a lead-poisoning survey of waterfowl. AB - Between 1983 and 1986 the National Wildlife Health Center (NWHC) conducted a nationwide study of lead poisoning of waterfowl from federal and state refuges. This survey was done to assist in identifying zones with lead-poisoning problems. One thousand forty one moribund or dead waterfowl were collected and examined. The presence or absence of 13 gross lesions selected as indicators of lead poisoning and three lesions indicating body condition was recorded. Lead poisoning diagnoses were based on the finding of at least 6-8 ppm (wet weight) lead in the liver and either lead shot in the gizzard content or at least one convincing gross lesion indicative of lead poisoning. Four hundred twenty-one of these waterfowl were diagnosed as lead poisoned. The NWHC survey provided a comprehensive basis for estimating the sensitivities, specificities, and likelihood ratios of the gross lesions of lead poisoning and the associated hepatic lead concentrations for several species of waterfowl. Some of the 13 defined gross lesions were more common than others; frequencies ranged from 3% to 80% in the 421 lead-poisoned waterfowl. The most reliable indicators of lead poisoning were impactions of the upper alimentary tract, submandibular edema, myocardial necrosis, and biliary discoloration of the liver. Each of the 13 lesions occurred more frequently in the lead-poisoned birds, but each of the lesions also occurred in waterfowl that died of other causes. The number of lead shot present in a bird's gizzard was only weakly correlated with its hepatic lead concentration; however, this weak correlation may have been adequate to account for differences in hepatic lead concentrations among species, once the weights of the species were taken into account. Although lead-poisoned ducks tended to have higher hepatic mean lead concentrations than did lead-poisoned geese or swans, the differences were probably a result of a greater dose of shot per body weight than to kinetic differences between species. Hepatic lead concentrations were independent of age and sex. Ninety-five percent of waterfowl diagnosed as lead poisoned had hepatic lead concentrations of at least 38 ppm dry weight (10 ppm wet weight). Fewer than 1% of the waterfowl that died of other causes had a concentration that high. This fifth percentile, of 38 ppm dry weight (10 ppm wet weight), is a defensible criterion for identifying lead-poisoned waterfowl when interpreting hepatic lead concentrations in the absence of pathological observations. PMID- 9732485 TI - Subacute and reproductive effects in mink from exposure to Fusarium fujikuroi culture material (M-1214) containing known concentrations of moniliformin. AB - This study was conducted to ascertain the subacute and reproductive effects in mink (Mustela vison) resulting from exposure to moniliformin, a toxic mycotoxin produced by Fusarium fungi. In a preliminary trial, adult mink were presented diets that contained targeted concentrations of 10, 20, 40, 80, 160, or 240 ppm moniliformin provided by F. fujikuroi culture material (M-1214). The mink fed diets that contained more than 40 ppm moniliformin refused to eat significant quantities of feed. Feeding adult mink diets that contained 8.1 or 17.0 ppm (wet weight) moniliformin, provided by F. fujikuroi culture material, in a 30-day subacute trial produced no significant adverse effects on feed consumption, body weights, hematologic parameters, or serum chemical values, and notable histologic changes in tissues that were examined. In the reproduction trial, female mink were exposed to the same dietary concentrations of moniliformin provided by F. fujikuroi culture material as in the subacute test from 2 weeks prior to the breeding season until their offspring (kits) were 8 weeks old. Consumption of the high-dose (17 ppm) diet resulted in significant neonatal mortality and reduced kit body weights at birth and at 8 weeks of age. Necropsy of 8-week-old kits from the control and high-dose groups revealed no gross or histologic lesions or alterations in liver, lung, or heart tissues that could account for the mortality observed in the kits exposed to the culture material. These results indicate that long-term (105-135 days) dietary exposure to F. fujikuroi culture material containing 17 ppm moniliformin is not lethal to adult female mink, but can have adverse effects on neonatal mink. PMID- 9732486 TI - Age-dependent accumulation of heavy metals in baikal seal (Phoca sibirica) from the Lake Baikal. AB - Concentrations of Fe, Mn, Zn, Cu, Cd, and Hg were determined in the liver, kidney, and muscle of 60 Baikal seals collected from Lake Baikal in 1992 to investigate age-dependent accumulation. Among essential elements, Fe concentrations in the muscle, liver, and kidney increased with age, suggesting development of diving ability. The concentrations of Mn, Zn, and Cu decreased with age, especially at immature stages. Toxic elements such as Hg and Cd decreased in adult males and thus the male-female difference was clearly observed in their concentrations, which differed from patterns usually found in marine mammals. Such accumulation patterns were due to difference in the feeding rates between males and females under low exposure to Hg and Cd. In addition, a greater excretion of Hg than that of Cd through molting and parturition was estimated. PMID- 9732487 TI - Lifetime health risk assessment from exposure of recreational users to polycyclic aromatic hydrocarbons. AB - In order to assess the lifetime risk of skin cancer for recreational users from dermal exposure to polycyclic aromatic hydrocarbons (PAHs), sediment samples were collected from beach sites along the St. Marys River near Sault Ste. Marie, Ontario, and in Hamilton Harbor and Toronto Harbor, Ontario, and analyzed for PAHs. Dermal exposure and lifetime skin cancer risk were estimated as follows: Concentrations of 11 PAHs with sufficient or limited evidence of carcinogenicity or mutagenicity were converted to benzo(a)pyrene (BaP) equivalents using toxic equivalency factors (TEFs). Lifetime dermal exposure values were derived based on the BaP equivalents in the silt + clay fraction taken as representative of suspended sediment particulates to which recreational users would be exposed. The lifetime health risk of skin cancer associated with such exposures was above the negligible risk level of 1.0 x 10(-6) at offshore Rytac, Lake George Channel, and Bell Point beaches in the St. Marys River; at Pier 4 Park in Hamilton Harbor; and at Humber Bay, Sunnyside Beach, Cherry Beach, and Water Rats Sailing Club in Toronto Harbor. Risk was negligible inshore at the Rytac and Bell Point beaches and at Squirrel Island and Ojibway Trailer Park along St. Marys River, at Lax Beach in Hamilton Harbor; and at Centre Island in Toronto Harbor. Strategies to reduce risk were developed with these communities; a key recommendation was to take a bath or shower within 24 h after a swim because virtually all the PAHs on the skin would be removed. PMID- 9732488 TI - Assessment of asbestos burden in the placenta and tissue digests of stillborn infants in South Texas. AB - The primary aim of this prospective study was to examine the tissues and placentas of autopsied stillborn infants for presence of asbestos fibers. Asbestos burden of lung, liver, skeletal muscle, and placenta digests of 82 stillborn infants was determined using standard bleach digestion technique. The digests were examined by electron microscopy, and the types of fibers determined using energy dispersive x-ray analysis and selected area diffraction analysis. Digests of 45 placentas collected from deliveries of liveborn healthy infants were processed and examined similarly as controls. Asbestos fibers were detected in 50% of the fetal digests and 23% of the placental digests of stillborn infants. Of the fibers present, 88% were chrysotile, 10% were tremolite, and 2% were actinolite and anthophyllite. Fibers measured 0.5-16.73 microgram in length (mean 1.55 microgram), and 0.03-0.8 microgram in width (mean 0.098 microgram). Lungs were most frequently positive for fibers (50%), followed by muscle (37%), placenta (23%), and liver (23%). Mean fiber counts were highest in the liver (58,736 f/g), followed by placenta (52,894 f/g), lungs (39,341 f/g), and skeletal muscle (31,733 f/g). Digests of 15% of the control placentas also showed asbestos fibers, although in very small numbers. The mean fiber count of the stillborn placentas (52,894 f/g) was significantly higher than the mean fiber count of the control placentas (mean 19 f/g) (p = 0.001). A highly significant association was found between fiber presence in stillborns and a maternal history of previous abortions (p = 0.007). A significant association was also found between fiber presence and placental diseases (p = 0.041). An association was suggested between working mothers and fiber presence (p = 0.19), although it did not reach statistical significance. The study documents the presence of small and thin asbestos fibers in stillborn fetal tissues and placenta. Significantly higher number of fibers were found in stillborn tissues compared to controls (liveborn placenta). The absence of a maternal history of asbestos-related occupations suggests that the fibers may have been acquired through environmental exposure. PMID- 9732489 TI - Exposure to methyl bromide during greenhouse fumigation on Crete, Greece. AB - In agricultural areas where greenhouses and dwellings are intermixed, the general population as well as the professional applicators may be exposed to pesticides. In a field study on Crete, exposure to methyl bromide during soil fumigation was assessed. Exposure of applicators (both contractors and farmers) were measured with personal air sampling equipment. Environmental monitoring inside and outside greenhouses combined with meteo data formed the basis for calculating the exposure of the general population with a computer aided dispersion model. Exposure of contractors exceeded the TLV value. The safe limit for the general population living close to a fumigated greenhouse is also exceeded. PMID- 9732490 TI - Costochondral junction fractures and intra-abdominal trauma in non-accidental injury (child abuse). AB - Rib fractures are a common skeletal manifestation of non-accidental injury (NAI) in infants and young children and are generally considered to be highly specific for abuse. There are, however, relatively few descriptions of fractures involving the costochondral junctions in NAI. We present three children (two boys, one girl; 7, 18, and 36 months of age) with anterior rib fractures which involved the sixth to ninth costochondral junctions. The fractures were bilateral in two children and symmetrical in one. They had appearances analogous to 'bucket handle' metaphyseal fractures of long bones. They were difficult to visualise and healed with minimal callus formation. These fractures were associated with major abdominal visceral injuries, which in themselves carry a significant morbidity and mortality. The importance of recognising such fractures is highlighted. PMID- 9732491 TI - MRI diagnosis of osteomyelitis of the cuboid bone in two infants. AB - We describe two infants in whom MRI diagnosed osteomyelitis of the cuboid bone when conventional X-rays were negative. Neoplastic, traumatic and ischaemic aetiologies could be excluded with the initial MR examinations. PMID- 9732493 TI - The impact of vesicoureteral reflux on contralateral renal length in infants with multicystic dysplastic kidney. AB - PURPOSE: The purpose of our study was to determine the influence of vesicoureteral reflux (VUR) on contralateral renal length in neonates and young infants with unilateral multicystic dysplastic kidney (MCDK). MATERIAL AND METHODS: We reviewed the imaging findings in 48 term neonates and infants (27 boys; 21 girls) who had unilateral MCDK (mean age at diagnosis 0.09 years; range 0-0.64 years). Each had renal ultrasonography (RUS), renal scintigraphy, and voiding cystourethrography before 1 year of age. The diagnosis of MCDK was based on characteristic imaging findings (i. e., an echogenic, cystic kidney at RUS that did not function at scintigraphy). None had contralateral hydronephrosis or cysts. We calculated an age-corrected z-score for contralateral renal length (at RUS) in each patient based on published standards. We examined the effects of gender, ipsilateral or contralateral VUR, and age at RUS on the contralateral renal length using multifactor ANOVA. RESULTS: Nine patients (19 %) had VUR into the contralateral kidney. The refluxing kidneys were significantly shorter (renal length: median 5.1 cm, mean 5.07 cm; z-score: median - 0.43, mean - 0.58) than the nonrefluxing kidneys (renal length: median 6.2 cm, mean 6. 08 cm; z-score: median 1.03, mean 1.04; P < 0.001). The contralateral kidney was more than 1 SD longer than the mean for age in none of the 9 patients with VUR on that side. By comparison, the contralateral kidney was more than 1 SD longer than the mean for age in 21 (54 %) of 39 patients with no VUR on that side, and more than 2 SD longer than the mean in 5 (13 %). CONCLUSION: VUR into the kidney contralateral to a MCDK is associated with smaller size of that kidney during the first year of life. PMID- 9732492 TI - Pelvis-shoulder dysplasia. AB - Pelvis-shoulder dysplasia is a rare focal skeletal dysostosis. We present the long-term follow-up of a patient with this condition. This patient has severe pelvic dysplasia but no involvement of the scapulae or clavicles. Despite the severity of the pelvic dysplasia, this man is able to function well. This is the fifth case of pelvis-shoulder dysplasia reported, but the only one documenting follow-up into adulthood. PMID- 9732494 TI - Asymmetric medullary nephrocalcinosis in two children. AB - Medullary nephrocalcinosis (MNC) is usually a bilateral process with symmetric involvement of both kidneys. Asymmetric medullary nephrocalcinosis has been previously reported in the literature, but has not been well illustrated or explained. We report the sonographic findings in two pediatric patients with hypercalcemia. In both patients an unrelated unilateral renal abnormality, (renal vein thrombosis in one and obstructive hydronephrosis in the other) prevented the development of MNC in the affected kidney, probably by decreasing the glomerular filtration rate and/or altering the renal tubular function. PMID- 9732495 TI - High-resolution CT findings in Wilson-Mikity syndrome: a case report. AB - Wilson-Mikity syndrome (WMS), an uncommon cause of respiratory distress presenting after birth, is radiologically characterised by varying degrees of interstitial thickening and bilateral cyst-like foci of hyperinflation. Aetiology and pathogenesis are still unknown. There are few reports of WMS in the paediatric literature and none describing the features and value of high resolution CT. The purpose of this report is to describe the radiographic findings and high-resolution CT appearance of WMS and to correlate them with the histopathological findings. PMID- 9732496 TI - Spontaneous correction of the malpositioned percutaneous central venous line in infants. AB - Malpositioning of the percutaneously placed central venous line (PCVL) or percutaneously inserted central catheter (PICC) in infants is not a rare occurrence. It has been occasionally observed that these lines spontaneously correct themselves. This prospective study was done to study the incidence of malposition and spontaneous correction. Using a modification of the standard method, 187 catheters were placed with 98.9 % success. Seven of these were initially malpositioned. All seven corrected themselves within a day when left in and used as a peripheral intravenous line. In many centers malpositioned catheters are taken out and replaced, which imposes great stress on the critically ill infant. Our study suggests that to avoid this stress the catheter should be left in place, since spontaneous correction may occur. PMID- 9732497 TI - Diagnostic imaging of primitive neuroectodermal tumour of the chest wall (Askin tumour). AB - OBJECTIVES: To describe the radiological features of primitive neuroectodermal tumour (PNET) of the chest wall (Askin tumour) at diagnosis and to analyse the radiological changes occurring as a consequence of treatment and during follow up. MATERIALS AND METHODS: Nine children with histologically proven PNET were studied. At diagnosis, all patients underwent chest X-ray (CXR), chest CT and bone scintigraphy; three patients also had MR and three had US. During treatment and follow-up, CT was performed in all patients. RESULTS: CT demonstrated a solid heterogeneous chest wall mass in all children at diagnosis and six had a rib lesion. Small nodular densities in the extra-pleural fat were identified in three patients at diagnosis. US, performed in three patients, excluded tumour infiltration of the lung or diaphragm, which had been suspected on CT. On MR, the lesions showed high signal intensity in T1-weighted/proton-density images and intermediate/high signal intensity in T2-weighted images compared with muscle. Minimal chest wall involvement was demonstrated in one case by MRI. Extensive necrosis of tumour mass with pseudo-cystic appearance was documented in the five patients who underwent chemotherapy. Macroscopically complete resection was performed in five patients but there was early local recurrence after surgery in two, identified by CT in one and by MR in the other. CONCLUSIONS: PNET of the chest wall should be considered in a child with a chest wall mass. CT is valuable for evaluating tumour extension at diagnosis, the effects of chemotherapy and assessing tumour recurrence after surgery. However, CT can overestimate pleural, lung or diaphragmatic infiltration, which are better evaluated by US. MR was superior to CT in the evaluation of tumour extension in one of three patients and may be considered complementary to CT, particularly in very large chest wall tumours. PMID- 9732498 TI - Glomerulocystic disease with hepatoblastoma in a neonate: a case report. AB - Glomerulocystic disease (GCD) is a very rare condition. Only two previous reports have linked this condition with hepatoblastoma. We report a neonate with US evidence of grossly enlarged echogenic kidneys and features typical of hepatic fibrosis, complicated by the presence of a hepatoblastoma. The report discusses the differential diagnosis and highlights GCD as one cause of large, bright kidneys on US. It also adds further evidence to the suggested association between GCD and hepatoblastoma. PMID- 9732499 TI - Bacterial orchitis in a baby with imperforate anus. PMID- 9732500 TI - Unusual gallbladder findings in two brothers with metachromatic leukodystrophy. AB - Characteristic biliary tree abnormalities in metachromatic leukodystrophy (MLD) include gallbladder polyposis and haemobilia. We report two brothers with MLD, who presented with uncommon biliary complications. One presented with gastric outlet obstruction secondary to gallbladder enlargement, which was treated by percutaneous aspiration. He later developed gallbladder carcinoma with liver metastases. His brother demonstrated US findings consistent with gallstones. PMID- 9732501 TI - MRI in the assessment of a newborn with cervical teratoma. AB - Teratoma of the head and neck is a rare lesion comprising 6 % of all teratomas, with only 3 % occurring in the cervical region [1]. Most are non-malignant lesions consisting of a variety of tissues of variable maturity, commonly with neuroepithelial and thyroid elements. They often present as a large cystic mass in the neck of a neonate or infant and frequently cause respiratory embarrassment due to local mass effect necessitating urgent surgical intervention. They may be difficult to distinguish from cystic hygromas, both clinically and radiologically. Imaging plays an important role in the assessment of these lesions, especially in preparation for surgery. We present a case of cervical teratoma and emphasise the role of MRI. PMID- 9732502 TI - Eosinophilic gastroenteritis mimicking idiopathic hypertrophic pyloric stenosis. AB - We report two infants with eosinophilic gastroenteritis (EG). This rare disease can mimic the clinical symptoms and US appearance of idiopathic hypertrophic pyloric stenosis (IHPS). US examination of the antropyloric region with a high frequency linear transducer can assist in the differentiation of EG from IHPS, which is important because the therapeutic approaches are completely different. Eosinophilic gastroenteritis should be considered in the differential diagnosis of IHPS, especially when there has been an ineffective pyloromyotomy. PMID- 9732503 TI - Systemic spread of meconium peritonitis. AB - Meconium peritonitis is a chemical peritonitis which occurs following bowel perforation during fetal life. It is generally looked upon as benign, resulting in no long-term sequelae. We present a case of a newborn infant with meconium peritonitis who developed infarcts in several organs. At autopsy the infarcts proved to be caused by emboli as a result of intravascular dissemination of meconium. To our knowledge, this is the first reported case of systemic spread of meconium peritonitis in the literature and suggests that meconium peritonitis may have more serious implications than generally thought. PMID- 9732504 TI - Malrotation in newborns following antenatal diagnosis of intra-abdominal cyst. AB - Two newborn girls had malrotation, small bowel in a subdiaphragmatic location on the right and leftward displacement of the liver. On antenatal scans, each had been diagnosed as having a large intra-abdominal cyst, but this had disappeared in both by the time of delivery. Both infants were asymptomatic at birth. One baby had a wrinkled abdominal wall, which is typically a component of prune belly syndrome. Both babies underwent Ladd procedure for their malrotation. In one, plate-like calcification over the hepatic capsule was the only residue of the previous cyst. In the other, mesenchymal hamartoma of the liver was diagnosed from histology of a collapsed adherent cyst. PMID- 9732505 TI - CT of splenic infarction in SLE. PMID- 9732506 TI - Craniosynostosis 1998: concepts and controversies. AB - The diagnosis of craniosynostosis requires attention to radiologic technique, familiarity with the signs of closure at each suture, and correlation with the clinical setting. Misdiagnosis may result in unwarranted calvarial or craniofacial surgery or a delay of surgery, necessitating a more extensive procedure. PMID- 9732507 TI - Intracranial lipoma of the quadrigeminal region associated with complex partial seizures. AB - A 7-year-old Japanese boy with an intracranial lipoma of the quadrigeminal region and complex partial seizures is reported. Among 28 published patients with lipoma originating in the quadrigeminal plate and ambient cistern, 6 suffered from seizures and 3 were mentally retarded. Our patient's seizures were controlled with carbamazepine. PMID- 9732508 TI - Disappearing suprarenal masses in fetuses and infants. PMID- 9732509 TI - Intrahepatic plexiform neurofibroma in neurofibromatosis 1. PMID- 9732510 TI - Salter-Harris type I fracture of the sacro-coccygeal joint. PMID- 9732511 TI - FORUM: Partnership Forum Framework: Participative Framework for Protected Area Outreach. AB - / Contemporary trends in natural resource management are reviewed, with specific reference to the shift in conservation management strategies away from law enforcement-based strategies towards strategies aimed at facilitating local community participation in the management of natural resources. This review lays a foundation for the presentation of a conceptual framework, the partnership forum framework, for the planning, implementation, and evaluationof protected area outreach programmes. The framework proposes that protected areas should function as integral components of the local social, economic, and environmental systems and that the integration of the protected area into these systems should be managed through comanagement institutions. The establishment of such institutions is discussed, and it is argued that the development of comanagement institutions can be characterized into four progressive phases: a preliminary communication phase, a problem-solving phase, a pilot project phase, and a comanagement phase. The framework proposes that during the three initial phases the partnership forum members will develop management procedures that they will use during the comanagement phase. The framework is presented as a design skeleton around which the site-specific characteristics of specific protected area outreach programs will combine to form an outreach program, i.e., the framework is process rather than project based.KEY WORDS: Sub-Saharan Africa; Integrated conservation and development PMID- 9732512 TI - PROFILE: "Low-Salt" Shrimp Aquaculture in Thailand: Goodbye Coastline, Hello Khon Kaen! AB - / Intensive shrimp culture has been confined to relatively narrow bands of land along the seashores of tropical developing nations due to the need for large volumes of saltwater for water exchange during the culture period. Recent developments in Thailand suggest, however, that this close association could soon be a thing of the past. Large numbers of Thai farmers are adopting low-salinity culture systems that rely upon sea or salt pan water that is trucked inland. This development greatly increases the potential for establishing shrimp cultivation much further from the coast than previously believed possible. The migration of intensive shrimp farming into freshwater environments, however, raises serious concerns over the disposal of pond effluents and the impact of saltwater intrusion on surrounding agricultural activities. In the absence of effective government regulation of the expansion and operation of the shrimp culture industry, supporting local nongovernmental organizations (NGOs) and community initiatives may be the only means of minimizing the negative impacts of shrimp farming on rural communities.KEY WORDS: Aquaculture; Shrimp; Salinity; Thailand PMID- 9732513 TI - Animal Burrowing Attributes Affecting Hazardous Waste Management. AB - / Animal burrowing is critical to the formation of soils and contributes to the interface between geological materials and organic life. It also influences the management of hazardous materials at nuclear waste facilities and elsewhere. For example, residues and waste products from the production of nuclear weapons are released onto the ground surface and within engineered burial structures. Soil bioturbation has exposed radionuclides and other hazardous materials to wind and rain, thereby risking inhalation and injury to humans and wildlife on and off site. Soil bioturbation can expand soil depths and spatial distributions of the source term of hazardous waste, potentially increasing chronic exposures to wildlife and humans over the long term. Ample evidence indicates that some of the large quantities of hazardous materials around the world have been released from soil repositories, where they have also contaminated and harmed biota. Key burrowing parameters influencing these outcomes include the catalog of resident species, and their abundance, typical burrow volumes (void space created by soil displacement), burrow depth profiles, maximum depth of excavation, constituents and structural qualities of excavated soil mounds, and proportion of the ground covered by excavated soil. Other important parameters include rate of mound construction, depth of den chambers, and volume of burrow backfill. Soil bioturbation compromised the integrity of some hazardous waste management systems using soil, but the environmental impact remains largely unknown. Designers and operators of waste management facilities, as well as risk assessors, need to understand how burrowing animals influence hazardous waste storage.KEY WORDS: Burrowing; Environmental impact; Radioactivity; Risk; Soil bioturbation; Hazardous waste PMID- 9732514 TI - Solid Waste Management in Nigeria: Problems and Issues. AB - / This paper is a presentation of the problems of solid waste management in Nigeria and certain important issues that must be addressed in order to achieve success. At the core of the problems of solid waste management are the absence of adequate policies, enabling legislation, and an environmentally stimulated and enlightened public. Government policies on the environment are piecemeal where they exist and are poorly implemented. Public enlightenment programs lacked the needed coverage, intensity, and continuity to correct the apathetic public attitude towards the environment. Up to now the activities of the state environmental agencies have been hampered by poor funding, inadequate facilities and human resources, inappropriate technology, and an inequitable taxation system. Successful solid waste management in Nigeria will require a holistic program that will integrate all the technical, economic, social, cultural, and psychological factors that are often ignored in solid waste programs.KEY WORDS: Solid waste; Management; Problems; Solutions; Nigeria PMID- 9732515 TI - RESEARCH: Environmental Racism in the Sunbelt? A Cross-Cultural Analysis. AB - / Sociologist Robert Bullard challenged the prevailing paradigm of environmentalism as a consensual issue in the United States by developing the concept of environmental racism. As he claims, ethnic minorities have been put "at greater environmental risk" than has the Caucasian majority in most areas of the country. This study of the Tucson metropolitan area examines this proposition by utilizing data from several sources: interviews with elected officials and other opinion leaders, GIS-generated socioeconomic data, articles in the press, and a literature review. We conclude that Bullard's concept has validity for this metropolitan area but that there also exist widely divergent differences of opinion on the subject. We explain why this is so. We further conclude that the allegation of "environmental racism" made by the Hispanic community in the 1980s and 1990s has had a transformative effect on local politics.KEY WORDS: Environment; Racism; Tucson; Garbage dumps; County government; Pollution; Equity; Justice PMID- 9732516 TI - Attitudes About Recreation, Environmental Problems, and Estuarine Health Along the New Jersey Shore, USA. AB - / Management of ecosystems has advanced by an improvement in our understanding not only of how ecosystems function, but of how people perceive their functioning and what they consider to be environmental problems within those systems. Central to such management is understanding how people view estuaries. In this article I explore the perceptions and attitudes of people about coastal recreation, environmental problems, and future land use along the New Jersey shore (USA) by interviewing people who attended a duck decoy and craft show on Barnegat Bay. The people who were interviewed engaged in more days of fishing than any other recreational activity and engaged in camping the least. There were significant differences in recreational rates as a function of gender and location of residence, with men hunting and fishing more than women and photographing less than women. Jet skis were perceived as the most severe environmental problem, with chemical pollution, junk, oil runoff and overfishing as second level problems. Birds were perceived as not an environmental problem at all. Fishing, hiking, preservation, and camping ranked as the highest preferred future land uses for the two sites examined (Oyster Creek Nuclear Generating Station, Naval Weapons Station Earle). The preferred future land uses for these two sites, which are not under consideration for land-use changes, were very similar to those of people living near the Department of Energy's Savannah River Site in South Carolina, despite the media attention and considerations of nuclear storage.KEY WORDS: Recreation; Perceptions; Environmental problem; Gender; Land use; Coastal PMID- 9732517 TI - Desire to Bargain and Negotiation Success: Lessons About the Need to Negotiate from Six Hydropower Disputes. AB - / We investigated the notion that successful negotiations require that all parties to the dispute must have a desire to bargain. This desire is most likely to be present when the dispute exhibits ripeness and each party believes a bargained solution is the most cost-effective way to resolve differences. Structured interviews of participants in six Federal Energy Regulatory Commission hydropower licensing consultations were conducted to determine the level of need to negotiate for each party. The findings indicate that a need to negotiate is a necessary, but not sufficient, condition for success. Several factors were associated with a need to negotiate: a weak BATNA (best alternative to a negotiated agreement); a salient issue; participants' sense of efficacy; a sense of inevitability; professional roles encouraging negotiation; and disputes about facts as opposed to disputes about values. Participants' need to negotiate fluctuated throughout the process and intensified when questions were ripe: i.e., critical issues were debated or the regulatory process required action.KEY WORDS: Alternative dispute resolution; Federal licenses; Federal Energy Regulatory Commission; Instream flow; Environmental planning PMID- 9732518 TI - Effect of Environmental Setting on Sediment, Nitrogen, and Phosphorus Concentrations in Albemarle-Pamlico Drainage Basin, North Carolina and Virginia, USA. AB - / Environmental settings were defined, through an overlay process, as areas of coincidence between categories of three mapped variables?Mland use, surficial geology, and soil drainage characteristics. Expert judgment was used in selecting factors thought to influence sediment and nutrient concentrations in the Albemarle-Pamlico drainage area. This study's findings support the hypothesis that environmental settings defined using these three variables can explain variations in the concentration of certain sediment and nutrient constituents. This finding underscores the importance of developing watershed management plans that account for differences associated with the mosaic of natural and anthropogenic factors that define a basin's environmental setting. At least in the case of sediment and nutrients in the Albemarle-Pamlico region, a watershed management plan that focuses only on anthropogenic factors, such as point-source discharges, and does not account for natural characteristics of a watershed and the influences of these characteristics on water quality, may lead to water quality goals that are over- or underprotective of key environmental features and to a misallocation of the resources available for environmental protection.KEY WORDS: Environmental setting; Water quality; Watershed management; Nutrients; Sediment PMID- 9732519 TI - Use of Airborne Thermal Imagery to Detect and Monitor Inshore Oil Spill Residues During Darkness Hours. AB - / Trials were conducted using an airborne video system operating in the visible, near-infrared, and thermal wavelengths to detect two known oil spill releases during darkness at a distance of 10 nautical miles from the shore in St. Vincent's Gulf, South Australia. The oil spills consisted of two 20-liter samples released at 2-h intervals, one sample consisted of paraffinic neutral material and the other of automotive diesel oil. A tracking buoy was sent overboard in conjunction with the release of sample 1, and its movement monitored by satellite relay. Both oil residues were overflown by a light aircraft equipped with thermal, visible, and infrared imagers at a period of approximately 1 h after the release of the second oil residue. Trajectories of the oil residue releases were also modeled and the results compared to those obtained by the airborne video and the tracking buoy. Airborne imagery in the thermal wavelengths successfully located and mapped both oil residue samples during nighttime conditions. Results from the trial suggest that the most advantageous technique would be the combined use of the tracking beacon to obtain an approximate location of the oil spill and the airborne imagery to ascertain its extent and characteristics.KEY WORDS: Airborne video; Thermal imagery; Global positioning; Oil-spill monitoring; Tracking beacon PMID- 9732521 TI - A Comparison of Urban-Proximate and Urban-Distant Wilderness Users on Selected Variables. AB - / The underlying premise of this study is that wilderness areas attract visitors desiring or expecting different wilderness experiences. In this study, wilderness areas were dichotomized according to distance from a large urban center (urban proximate vs urban-distant). Four wilderness areas in southern California were used as the study sites. Comparisons were made on selected attributes commonly associated with the wilderness experience. Differences were observed on a number of variables such as acceptable number and type of encounters with other visitors, management preferences, and preferred group sizes. The findings of this study are congruent with those from previous studies and suggest that distance to large urban centers may be a functional variable in explaining differences among selected wilderness attributes.KEY WORDS: Expectancy theory; Normative standards; Wilderness; Wilderness experience; Urbanization PMID- 9732520 TI - Effects of Metal Mining and Milling on Boundary Waters of Yellowstone National Park, USA. AB - / Aquatic resources in Soda Butte Creek within Yellowstone National Park, USA, continue to be threatened by heavy metals from historical mining and milling activities that occurred upstream of the park's boundary. This includes the residue of gold, silver, and copper ore mining and processing in the early 1900s near Cooke City, Montana, just downstream of the creek's headwaters. Toxicity tests, using surrogate test species, and analyses of metals in water, sediments, and macroinvertebrate tissue were conducted from 1993 to 1995. Chronic toxicity to test species was greater in the spring than the fall and metal concentrations were elevated in the spring with copper exceeding water quality criteria in 1995. Tests with amphipods using pore water and whole sediment from the creek and copper concentrations in the tissue of macroinvertebrates and fish also suggest that copper is the metal of concern in the watershed. In order to understand current conditions in Soda Butte Creek, heavy metals, especially copper, must be considered important factors in the aquatic and riparian ecosystems within and along the creek extending into Yellowstone National Park.KEY WORDS: Mining; Metals; Toxicity; Biomonitoring; Copper; Yellowstone National Park PMID- 9732522 TI - Sources and Distribution of Polychlorinated Terphenyls at a Major US Aeronautics Research Facility. AB - / High concentrations of an unusual, complex mixture of chlorinated compounds were discovered in sediments and oysters near a federal aeronautics facility during implementation of a pollutant screening protocol. The mixture was identified as Aroclor 5432, a polychlorinated terphenyl (PCT) formulation, produced in the US until 1972. PCTs, particularly low chlorinated mixtures, have rarely been reported in the environment, despite significant manufacture and usage. Releases were traced to two outfalls. Creek sediments downstream of one contained concentrations as high as 200,000 |gmg/kg (dry weight basis); those in indigenous oysters reached 35,000 |gmg/kg, indicating significant bioavailability and bioaccumulation potential. Subsequent work showed that PCTs were widely disseminated in marsh grass, crabs, and fish. PCTs, PCBs, and mercury were also detected in storm drain lines entering these outfalls. The lines received input from both storm water and research buildings. Historical hydraulic fluid leaks and in-service compressor fluids in some buildings contained PCTs and PCBs. Contaminated materials on-site were removed to minimize pollutant spread. Aroclor 5432 usage, most likely as compressor/hydraulic fluid additives, probably ended about ten years prior to its on-site detection. In terms of biological effects, intraperitoneal injection of fish with Aroclor 5432 induced cytochrome P-4501A (CYP1A) and ethoxyresorufin O-deethylase (EROD) activity to a similar degree as PCB Aroclor 1254 and to a greater extent than PCT Aroclor 5460. The presence of high concentrations of PCTs contributed to the facility being included on the National Priorities List. It subsequently became the first US federal facility to sign a Federal Facility Agreement, identifying cleanup responsibilities, prior to formal listing.KEY WORDS: Polychlorinated terphenyls; Aroclor; Contaminated sediments; Hydraulic fluid; Enzyme induction; Polychlorinated biphenyls PMID- 9732524 TI - Title Index. PMID- 9732523 TI - ENVIRONMENTAL AUDITING: Indicators Assessment for Habitat Conservation Plan of Yolo County, California, USA. AB - / Whereas habitat conservation plans (HCPs) have been intended to provide comprehensive environmental mitigation for multiple species, they often narrow in focus to one species and either one mitigation site or unspecified sites. We developed an indicators framework from which to rate land units for their ecological integrity, collateral values (nonbiological qualities that can improve conservation), and restoration and conservation opportunities. The ratings of land units were guided by the tenets of conservation biology and principles of landscape and ecosystem ecology, and they were made using existing physical and floral information managed on a GIS. As an example of how the indicators approach can be used for HCPs, the 29 legally rare species targeted by the Yolo County HCP were each associated with vegetation complexes and agricultural crops, the maps of which were used for rating some of the landscape indices. The ratings were mapped so that mitigation can be directed to the places on the landscape where the legally rare species should benefit most from conservation practices. The most highly rated land units for conservation opportunity occurred along streams and sloughs, especially where they emerged from the foothills and entered the Central Valley and where the two largest creeks intersected the Sacramento River flood basin. We recommend that priority be given to mitigation or conservation at the most highly rated land units. The indices were easy to measure and can be used with other tools to monitor the mitigation success. The indicators framework can be applied to other large-area planning efforts with some modifications.KEY WORDS: Ecosystem; Indicators; Landscape; Mitigation; Planning; Yolo County; California PMID- 9732526 TI - A chitinase encoding gene (chit1 gene) from the entomopathogen Metarhizium anisopliae: isolation and characterization of genomic and full-length cDNA. AB - There are no reports to date of entire gene sequences coding for chitinolytic enzymes from entomopathogenic fungi, even though these enzymes act synergistically with proteolytic enzymes to solubilize insect cuticle during the key step of host penetration, having considerable importance in the biological control of some insect pests. This paper reports the complete nucleotide sequence and analysis of the chromosomal and full-length cDNA copies of the regulated gene (chit1) coding one of the chitinases produced by the biocontrol agent Metarhizium anisopliae. Degenerated primers, encompassing conserved regions of other fungal chitinases, were used to amplify a 650-bp DNA fragment, which was used to isolate genomic and cDNA clones from M. anisopliae. Albeit at least two different chitinases are characterized in this fungus, only one chit gene was isolated. The chit1 gene is interrupted by three short typical fungal introns and has a 1,521 bp ORF, which encodes a protein of 423 amino acids with a stretch of 35 amino acid residues displaying characteristics of signal peptide. The deduced sequence of the mature protein predicts a 42-kDa protein with pI of 5.8. Southern analysis of genomic DNA indicates a single copy of chit1 in the M. anisopliae genome. PMID- 9732527 TI - Changes in protein synthesis as a consequence of heme depletion in Escherichia coli. AB - Two-dimensional gel electrophoresis and N-terminal amino acid sequence determination were used to compare the protein synthesis of exponentially growing Escherichia coli with heme-deficient cells. Mutation of the E. coli hemA gene encoding glutamyl-tRNA reductase resulted in the absence of detectable amounts of heme. As a consequence of heme deficiency, the induction of tryptophanase (trpA), citrate synthase (gltA), and aldehyde dehydrogenase (aldA) and the repression of enolase (eno) and phosphoglycerate kinase (pgk) were observed. All induced genes are under the control of the catabolite repressor protein Crp. The observed changes in gene expression as a consequence of heme depletion are discussed. PMID- 9732528 TI - Overexpression of a mutant B subunit in toxigenic Vibrio cholerae diminishes production of active cholera toxin in vivo. AB - A mutant cholera toxin B subunit containing a G33E substitution was constructed and expressed in V. cholerae. The G33E amino acid substitution did not affect the amount of recombinant CTB secreted to the culture medium. The overexpression of the mutant B subunits in wild-type toxigenic cholera vibrios led to an 80% decrease in production of active cholera toxin in vitro and in vivo. Overexpression of BG33E subunits could be instrumental in the increase of the biosafety of live attenuated cholera candidate vaccine strains. PMID- 9732529 TI - Molecular analysis of chitin synthase 1 (CHS1) gene sequences of Trichophyton mentagrophytes complex and T. rubrum. AB - Nucleotide sequences of chitin synthase 1 (CHS1) gene of dermatophytes, Arthroderma benhamiae, A. simii, A. vanbreuseghemii, Trichophyton mentagrophytes var. interdigitale (T. interdigitale), and T. rubrum were analyzed for their phylogenetic relationship. About 620-bp genomic DNA fragments of the CHS1 gene were amplified from these dermatophytes by polymerase chain reaction (PCR) and sequenced. The CHS1 nucleotide sequences of these five dermatophytes showed more than 90% similarity between the species. The phylogenetic analysis of their sequences revealed that A. benhamiae, A. simii, A. vanbreuseghemii, and T. rubrum were genetically distinct from one another, but T. interdigitale was genetically very close to A. vanbreuseghemii. On the other hand, a specific restriction endonuclease site of HinfI was present in the CHS1 gene fragment of T. rubrum but not in those of A. benhamiae, A. simii, A. vanbreuseghemii and T. interdigitale. The molecular analysis of CHS1 genes will provide useful information for the identification of these Trichophyton species and the understanding of their evolution. PMID- 9732530 TI - In vivo production of neuraminidase by Pasteurella haemolytica in market stressed cattle after natural infection. AB - Pasteurella haemolytica (Ph) is the most important cause of the bovine acute fibrinohemorrhagic pneumonia that occurs in market stressed calves after shipment to feedyards. Recent characterization of neuraminidase production by these organisms has shown that all 16 serotypes produce an immunologically similar form of the enzyme. Anti-neuraminidase antibody against PhA1 and PhA6 was determined in 101 2- to 5-month-old calves, on their farms of origin, at the order buyer barn (OBB), and through 28 days in the feedyard. Half of the calves were vaccinated with a killed Ph serotype-A1 (PhA1) product. Nasal secretion and tonsil wash specimens were cultured for Ph and Pasteurella multocida (Pm). Serum antibody against PhA1 and PhA6 was measured by indirect hemagglutination (IHA), and anti-neuraminidase antibody was determined by the neutralization assay. At the feedyard, 73 calves had respiratory tract disease. IHA values ranged between 1:2 and 1:1024 for PhA1 and between 1:2 and 1:512 for Ph serotype A6 (PhA6). Forty-two, 24, and 28% of the calves were infected with PhA1, PhA6, and Pm, respectively. Ninety-six percent of the calves experienced an increase in anti PhA1 neuraminidase antibody when sera drawn on feedyard day 28 were compared with sera drawn on the farm. These data demonstrate that the enzyme neuraminidase is produced in vivo in market stressed cattle after a natural Ph infection. PMID- 9732531 TI - Localization of putative virulence genes on a physical map of the bacillus thuringiensis subsp. gelechiae chromosome AB - The insect pathogen Bacillus thuringiensis (Bt) has earlier been shown to possess virulence factors in addition to the crystal toxins. Bt subsp. gelechiae strain Bt13 lacks crystals but is still virulent to lepidopteran insects. Among the virulence co-expressed genes are two phospholipases; phosphatidylinositol specific phospholipase C (PI-PLC) and phosphatidylcholine-degrading phospholipase C (PC-PLC), flagellin, and beta-lactamase I. In addition to these putative virulence factors the toxic neutral metalloprotease immune inhibitor A (InA) has been identified. In this paper we report a circular 5.9 Mb combined physical and genetic map of the of the Bt subsp. gelechiae chromosome. The genes encoding PI PLC, PC-PLC, InA, flagellin, and beta-lactamase I are shown to be scattered over the chromosome. The PLC-encoding genes have been cloned from Bt13, and DNA sequencing showed that the Bt subsp. gelechiae PLC genes are >90% identical to their previously cloned equivalents from Bt or B. cereus. An HD-1 crystal toxin (cryIA) gene probe was found to hybridize to the Bt13 chromosome, but not to extrachromosomal elements. PMID- 9732532 TI - Partial purification and characterization of ornithine carbamoyl transferase (OCT) from the cyanobacterium nostoc sp. Strain PCC 73102 AB - Ornithine carbamoyl transferase (OCT) catalyzes the formation of citrulline and orthophosphate from ornithine and carbamoyl phosphate. We have partially purified OCT from the filamentous cyanobacterium Nostoc sp. strain PCC 73102, using ammonium sulfate precipitation (35-55%), a gel-filtration column (Sephacryl S 200), followed by an affinity column (Sepharose-6B-PALO). The partially purified OCT was analyzed on native-PAGE and shown to be an active enzyme with an estimated molecular weight of approximately 80 kDa. The isoelectric point was determined to be about 6.2. Varying the ornithine concentration resulted in a hyperbolic response of the reaction velocity at lower concentrations. Ornithine concentrations above 2 mM inhibited the enzyme. A hyperbolic response of the OCT reaction was observed when increasing the carbamoyl phosphate concentration. From a double reciprocal plot, a saturation concentration of 0.8 mM and a Vmax of 0.4 U/mg may be calculated. None of the tested compounds (argininosuccinate, arginine, aspartic acid, urea) had any significant positive effect on the in vitro activity of the partially purified OCT. Moreover, at concentrations higher than 10 mM, all tested compounds had an inhibitory effect. PMID- 9732533 TI - Algicidal effect of peridinium bipes on microcystis aeruginosa AB - Peridinium bipes exerted an inhibitory effect on the growth of Microcystis aeruginosa. The algicidal action of water-soluble extract from P. bipes was studied. After treatment with P. bipes extract, the absorption spectrum of M. aeruginosa culture changed markedly, particularly in the ranges of 500 approximately 650 and 420 approximately 460 nm. An increase in absorption in this wavelength resulted from a leakage of phycobilines, both phycocyanin and allophycocyanin, from the treated cells. However, no leakage of chlorophyll was detected. The leakage of phycobilines was a short-term effect, detectable within 1 h of incubation. Both the plasmalemma and thylakoid membranes in M. aeruginosa cells were damaged and distorted in fine structure after treatment with P. bipes extract. It is assumed that algicide from P. bipes exerts its effect on the cell membranes, giving rise to changes in membrane permeability and a dissociation of phycobiline assemblages, but not of chlorophyll complex, on the thylakoid membranes. As a result, phycobilines were leaked out from the cells. PMID- 9732534 TI - Analysis of the botulinum neurotoxin type F gene clusters in proteolytic and nonproteolytic Clostridium botulinum and Clostridium barati. AB - Comparison of genes encoding type F botulinum neurotoxin progenitor complex in strains of proteolytic Clostridium botulinum strain Langeland, nonproteolytic Clostridium botulinum strain 202F, and Clostridium barati strain ATCC 43256 reveals an identical organization of genes encoding a protein of molecular mass of approx. 47 kDa (P-47), nontoxic-nonhemagglutinin (NTNH) and botulinum toxin (BoNT). Although homology between the protein components of the complexes encoded by these different species all producing botulinum neurotoxin type F is considerable (approx. 69-88% identity), exceptionally high homology is observed between the C-termini of the P-47s (approx. 96% identity) and the NTNHs (approx. 94% identity) encoded by Clostridium botulinum type F strain Langeland and Clostridium botulinum type A strain Kyoto. Such a region of extremely high sequence identity is strongly indicative of recombination in these strains synthesizing botulinum neurotoxins of different antigenic types. PMID- 9732535 TI - Gene organization and transcriptional analysis of the Spiroplasma citri rpsB/tsf/x operon. AB - The nucleotide sequence of a 6863-bp Spiroplasma citri DNA fragment comprising the spiralin gene was determined. Sequence analysis revealed eight putative ORFs that encode ribosomal protein S2, elongation factor Ts, spiralin, 6 phosphofructokinase, pyruvate kinase, and three unidentified proteins (A, B, and X). The gene organization reported here is different from that previously published. Northern blot analysis of rpsB, tsf, and x transcripts indicates that these genes are organized into a single transcriptional unit (operon). However, the detection of an additional transcript corresponding to the rpsB gene alone suggests that a transcriptional mechanism should occur in the 3' region of the rpsB gene, allowing a conditional transcription termination. PMID- 9732536 TI - Methods for detection of conjugative plasmid transfer in aquatic environments. AB - Donor and recipient counter selection was evaluated by selecting bacteria that received plasmid RP4 by conjugation on filters and in lake water microcosms. Three counter selection systems were compared; (i) Use of antibiotic-resistant recipients, (ii) use of an auxotrophic donor, and (iii) use of a donor with chromosomal suicide genes. Transfer efficiencies of transconjugants per recipient obtained with the three different counter selection systems in filter-matings were not significantly different. Some nalidixic acid-resistant recipients became partly sensitive to nalidixic acid after receiving the plasmid. Use of an auxotrophic donor was a feasible and easy way to recover indigenous transconjugants. A strain with two copies of the suicide gene gef was successfully eliminated in filter-matings, but elimination of the donor in microcosms by induction of the suicide genes did not succeed. Thus, this counter selection system was not usable in microcosm experiments. PMID- 9732537 TI - Physiological and molecular biological characterization of ammonia oxidation of the heterotrophic nitrifier Pseudomonas putida. AB - The heterotrophic nitrifier Pseudomonas putida aerobically oxidized ammonia to hydroxylamine, nitrite, and nitrate. Product formation was accompanied by a small but significant release of NO, whereas N2O evolution could not be detected under the assay conditions employed. The isolate reduced nitrate to nitrite and partially further to NO under anaerobic conditions. Aerobically grown cells utilized gamma-aminobutyrate as a carbon source and as a N-source by ammonification. The physiological experiments, in particular the inhibition pattern by C2H2, indicated that P. putida expressed an ammonia monooxigenase. DNA hybridization with an amoA gene probe coding for the smaller subunit of the ammonia monooxigenase of Nitrosomonas europaea allowed us to identify, to clone, and to sequence a region with an open reading frame showing distinct sequence similarities to the amoA gene of autotrophic ammonia oxidizers. PMID- 9732538 TI - News & notes. Efficient library construction with a TA vector and its application to cloning of the phytoene synthase gene from the cyanobacterium Spirulina platensis. AB - An efficient and simple method for constructing a genomic DNA library is presented by use of a TA cloning vector. It is based on sonicative cleavage of genomic DNA and modification of the fragment ends with Taq DNA polymerase, followed by ligation with a TA vector. This method was successfully applied to cloning of the phytoene synthase gene crtB from Spirulina platensis. The method is useful when the genomic DNA is not well digested with restriction enzymes owing to methylation or other reasons. PMID- 9732539 TI - Primitive neuroectodermal tumors. AB - The primitive neuroectodermal tumors (PNETs) of Hart and Earle constitute an important fraction of pediatric brain tumors that are clinically characterized by their aggressive behavior. In 1983, Rorke expanded the term "PNET" to include all small cell embryonal neoplasms of neuroectodermal origin, regardless of the location of the tumor. More recently, Dehner also proposed the terms "central" and "peripheral" PNETs, and the provocative concept of "PNET" has now come to encompass a diverse group of tumors in both the central and peripheral nervous systems. The acronym "PNET" has thus become a confusing and controversial term in the diagnosis and classification of pediatric embryonal tumors. We reviewed ten papers concerning the various aspects of PNETs. PMID- 9732540 TI - Are p53 mutations and p53 overexpression prognostic factors for astrocytic tumors? AB - The p53 tumor supressor gene is the most frequently mutated gene in human cancers. Approximately 40% of astrocytic tumors have alterations of the p53 gene, which are considered to play an important role in tumorigenicity and malignant progression. Since the main functions of normal p53 are cell-cycle regulation and induction of apoptosis, p53 mutations are considered to be associated with rapid tumor growth and resistance to radiation and chemotherapy. Although in certain cancers, p53 mutations are considered a poor prognostic factor, the predictive role in astrocytic tumors is not clear. Immunohistochemical studies have shown conflicting results, probably because this technique fails to provide a reliable p53 gene status. Mutation analyses have shown the association between p53 mutations and shorter survival of high-grade gliomas only in pediatric patients, but not in adults. This may suggest that p53 mutations have a relatively lower impact on the survival of malignant astrocytomas than other gene alterations, which pediatric tumors rarely harbor. PMID- 9732541 TI - Hypothermia before and after insult. AB - It is well established that hypothermia will protect cerebral and cord tissue against necrosis from prolonged ischemia. The optimum depth of hypothermia and safe total time has not yet been established. There is considerable evidence that hypothermia may be useful as a treatment adjunct after trauma or ischemias, but the same caveats apply. There are agreed risks, the exact nature of which are unknown, and there are suspicions that hypothermia after the insult may be only delaying the natural recovery processes. PMID- 9732542 TI - Imaging in epidural lipomatosis. AB - This paper reviews the literature concerning symptomatic epidural lipomatosis. The different diagnostic procedures (conventional myelography, computed tomography, and magnetic resonance imaging) and therapeutic approaches (including medical treatment, surgery, and dietary influence) are discussed. PMID- 9732543 TI - Evaluation of surgical treatment outcome in epilepsy. AB - Seven recent papers are reviewed for outcomes following epilepsy surgery. The criteria of outcome assessment are analyzed and compared. All studies agree in indicating that the combination of the classic evaluation of seizure frequency with that of quality of life is required for a comprehensive view of the surgical outcome. However, the assessment modalities and outcome scales proposed present relevant differences. The need for standardization is apparent. A surgical outcome scoring system using multiple measures is recommended. The outcome evaluation should be performed not sooner than 2 years after surgery. The most relevant of the many variables of the outcome should be selected in such a way as to permit assessment of the epileptological response to surgery as well as the changes in the quality of life. PMID- 9732544 TI - Interventional neuroradiology. AB - The different interventional neuroradiological procedures are continually developing. New fields such as fibrinolytic therapy or intracranial stent treatment of vascular stenosis have become the object of interest. The analysis of the results of intravascular procedures has become more refined, with a critical evaluation of the quality of results and complication rates. A more solid scientific evaluation of these procedures is taking the place of the intial enthusiasm, offering a better basis for patient selection for neuroradiological procedures. In this analysis we present papers dealing with complications, analysis and treatment; results (follow-up of giant aneurysms treatment); use of stent in the internal carotid artery territory; and intra-arterial thrombolytic therapy (results and patient selection). PMID- 9732545 TI - Clinical experience with endovascular treatment of aneurysms using Guglielmi detachable coils. AB - Guglielmi detachable coils (GDCs) provide an endovascular means for aneurysm treatment; however, their role has yet to be defined. This article reviews the most recent clinical series regarding efficacy, safety, and clinical outcomes in both the acute and nonacute setting. Successful treatment was possible in the majority of cases and included cases of complete aneurysm obliteration and with only a small neck remnant. Patients with a neck remnant often received additional treatments although some patients went on to complete obliteration without further treatment. Successful treatment provided protection from rebleeding in a follow-up period of 2 years. Success depended on operator experience as well as width of the aneurysm ostium. Recurrence and incomplete obliteration were more common with giant or large aneurysms and aneurysms with a wide base. Major procedure-related complications predominantly resulted from intraprocedural rupture and thromboembolic events. These could both be treated via endovascular means at the time of the procedure. Thromboembolic events occurred more frequently with acutely ruptured aneurysms, especially aneurysms with a wide base. Vasospasm rates were not found to vary significantly from those found in surgical series when corrected for Fisher grouping. Morbidity and mortality rates as well as Glasgow outcome scores were at least as good as what would be expected from surgery during both the acute and nonacute setting. The papers reviewed indicate that the GDC provides safe and efficacious treatment for most berry aneurysms in both the acute and nonacute setting relative to surgical results. PMID- 9732546 TI - Role of chemotherapy in the treatment of low-grade gliomas. AB - Being the most prevalent group of brain tumors in all age groups, low-grade gliomas still leave the question of the optimal treatment open. Despite the indolent, non-aggressive behavior of these tumors, there has been no dramatic improvement in treatment results. Surgery and radiotherapy have certain limitations in the treatment of patients with low-grade gliomas. The purpose of this review is to display a role of chemotherapy in overcoming these therapeutic obstacles. The recent reports of groups studying treatment for low-grade gliomas are analyzed. PMID- 9732549 TI - Publications scanned for pertinent articles. PMID- 9732548 TI - Papers reviewed in this issue. PMID- 9732550 TI - Patterns of embryonic neurogenesis in a primitive wingless insect, the silverfish, Ctenolepisma longicaudata: comparison with those seen in flying insects. AB - Neurogenesis was examined in the central nervous system of embryos of the primitively wingless insect, the silverfish, Ctenolepisma longicaudata, using staining with toluidine blue (TB) and the incorporation of bromodeoxyuridine (BUdR). The silverfish has the same number and positioning of neuroblasts as seen in more advanced insects and the relative order in which the different neuroblasts segregate from the neuroectoderm is highly conserved between Ctenolepisma and the grasshopper, Schistocerca. Of the 31 different neuroblasts found in a thoracic segment, one (NB 6-3) has a much longer proliferative period in silverfish. Of the remainder, 14 have similar proliferative phases, while16 neuroblasts have extended their proliferative period by 10% of embryogenesis or greater in the grasshopper as compared with the silverfish. Both insects had similar periods of abdominal neurogenesis except that in the silverfish terminal ganglion a prominent set of neuroblasts continued dividing until close to hatching, possibly reflecting the importance of cercal sensory input in this insect. This comparison between silverfish and grasshopper shows that the shift from wingless to flying insects was not accompanied by the addition of any new neuronal lineages in the thorax. Instead, selected lineages underwent a proliferative expansion to supply the additional neurons presumably needed for flight. The expansion of specific thoracic lineages was accompanied by the reduction of the terminal abdominal lineages as flying insects began to de emphasize their cercal sensory system. PMID- 9732551 TI - Innervation regulates the metamorphic fates of larval abdominal muscles in the moth, Manduca sexta. AB - With the onset of metamorphosis, the abdominal muscles of the moth, Manduca sexta, follow one of three developmental fates: maintenance, respecification, or death. The maintained muscles retain their larval size and morphology throughout adult development. The respecified and dying muscles dedifferentiate, which involves regression, nuclear degeneration, and myofibril breakdown. Nuclei in both dying and respecified muscles also proliferate. The amount of nuclear degeneration is greater in the dying muscle fibers, and the amount of nuclear proliferation is greater in the respecified muscles. Four to ten days after pupation, the sizes of the respecified muscles stabilize while the dying muscles are lost. During regression, a subset of the respecified muscle fibers die. The surviving respecified muscle fibers grow and differentiate during the last half of adult development. In respecified muscles, denervation triggers an increased amount of nuclear degeneration and a decreased amount of nuclear proliferation. As a result, denervated respecified fibers experience increased muscle regression including an increased loss of muscle fibers and sometimes muscle death. Surviving respecified fibers still grow and differentiate yet are only 5 to 12% of the control size. Denervation triggers dedifferentiation in maintained muscles, resulting in fiber loss and occasionally muscle death. The percentage of fibers which dedifferentiate varies between different muscles. Denervation also triggers nuclear proliferation, with the amount of nuclear proliferation correlated with the extent of dedifferentiation of the individual muscle fibers. The dedifferentiated maintained fibers subsequently undergo differentiation in the absence of muscle growth. PMID- 9732552 TI - Combinatorial control of Drosophila mef2 gene expression in cardiac and somatic muscle cell lineages. AB - The Drosophila mef2 gene encodes a MADS domain transcription factor required for the differentiation of cardiac, somatic, and visceral muscles during embryogenesis and the patterning of adult indirect flight muscles assembled during metamorphosis. A prerequisite for D-MEF2 function in myogenesis is its precise expression in multiple cell types during development. Novel enhancers for D-mef2 transcription in cardiac and adult muscle precursor cells have been identified and their regulation by the Tinman and Twist myogenic factors have been demonstrated. However, these results suggested the existence of additional regulators and provided limited information on the specification of progenitor cells for different muscle lineages. We have further characterized the heart enhancer and show it is part of a complex regulatory region controlling the activation and repression of D-mef2 transcription in several cell types. The mutation of a GATA sequence in the enhancer changes its specificity from cardial to pericardial cells. Also, the addition of flanking sequences to the heart enhancer results in expression in a new cell type, that being the founder cells of a subset of body wall muscles. As tinman function is required for D-mef2 expression in both the cardial and founder cells, these results define a shared regulatory DNA that functions in distinct lineages due to the combinatorial activity of Tinman and other factors that work through adjacent sequences. The analysis of D-mef2-lacZ fusion genes in mutant embryos revealed that the specification of the muscle precursor cells involved the wingless gene and the activation of a receptor tyrosine kinase signaling pathway. PMID- 9732553 TI - H19 and Igf2 are expressed and differentially imprinted in neuroectoderm-derived cells in the mouse brain. AB - Igf2 and H19 are reciprocally imprinted genes that are closely linked and coexpressed in tissues of mesodermal and endodermal origin. Here we report that coexpression of these genes is also found in specific fetal tissues of neuroectodermal origin, that is in the ventral midline region of both the hindbrain and spinal cord. For cells of neuroectodermal origin, complete absence of Igf2 and H19 transcription was previously described. Analysis of allele specific expression of both Igf2 and H19 in the ventral midline region of the hindbrain shows that H19 is expressed monoallelically, with the paternal allele being silent, whereas Igf2 is expressed biallelically. Furthermore, we observed a strong influence of the parental species background, in that the Mus musculus allele was always expressed at higher levels than the M. spretus allele. This was observed when the M. spretus allele was contributed by the mother or by the father. An analysis of Igf2 methylation by bisulphite genomic sequencing provided no clear answer as to whether Igf2 expression and methylation are linked in a tissue of neuroectodermal origin. Taken together, our results provide novel information on H19 and Igf2 expression and imprinting patterns in the fetal mouse brain. In addition, they indicate that some aspects of Igf2 regulation in cells of neuroectodermal origin do not follow the pattern that exists in mesoderm- and endoderm-derived tissues. Apart from the ventral midline region, H19 and Igf2 were found to be coexpressed in the ectodermally derived Rathke's pouch and in some circumventricular organs of the brain, such as the organum vasculosum of the lamina terminalis (OVLT) and the pineal gland. PMID- 9732554 TI - Expression of SC1 is associated with the migration of myotomes along the dermomyotome during somitogenesis in early mouse embryos. AB - SC1 is a secreted glycoprotein with a high amino acid sequence similarity to SPARC (Secreted Protein, Acidic, Rich in Cysteine). SC1 transcripts were first detected in mouse embryos after day 8.5 post coitus (p.c.) in somites at the medial lip of the dermomyotome. Expression of SC1 transcripts by the progenitor cells continued as they began involuting under the dermomyotome and during their migration along the lateral wall of the dermomyotome. After myotome migration was completed, SC1 mRNA expression was downregulated in the trunk region. The data indicate that SC1 expression is restricted to the initial stages of epaxial myotome differentiation and migration, undergoing rapid downregulation prior to myotome emigration from the somitic environment. PMID- 9732556 TI - 4D confocal microscopy of Dictyostelium discoideum morphogenesis and its presentation on the Internet. AB - Methods to present three-dimensional (3D) and time series of 3D datasets (4D) are demonstrated using the recent advances in confocal microscopy and computer visualization. The process of cell sorting during tip formation in the slime mould Dictyostelium discoideum is examined as an example by in vivo confocal microscopy of spectrally different green fluorescent protein (GFP) variants as reporters of cell-type specific gene expression. Also, cell sorting of the co aggregating slime mould species D. discoideum and D. mucoroides is observed using a GFP variant and a spectrally distinguishable fluorescent vital stain. The confocal data are handled as 3D and 4D datasets, their processing and the advantages of different methods of visualization are discussed step by step. Selected sequences of the experiments can be viewed on the Internet, giving a much better impression of the complex cellular movements during Dictyostelium morphogenesis than printed photographs. PMID- 9732555 TI - A novel member of the bombyxin gene family: structure and expression of bombyxin G1 gene, an insulin-related peptide gene of the silkmoth Bombyx mori. AB - Bombyxin G1 gene, a novel insulin-related peptide gene of the silkmoth Bombyx mori, has been identified. The G1 gene encodes a precursor peptide which shows 41 56% and 28% sequence identities with preprobombyxins previously characterized and human preproinsulin, respectively. The G1 gene forms a pair with bombyxin C2 gene with opposite transcriptional orientation in a bombyxin gene cluster. The bombyxin G1 mRNA in Bombyx brain was shown to locate in four pairs of medial neurosecretory cells. PMID- 9732557 TI - Preventive orthopedics. PMID- 9732558 TI - Consistency of lumbar discograms of the same disc obtained twice at a 2-week interval: influence of needle tip position. AB - : Although numerous papers have emphasized the importance of accurate needle positioning in lumbar discography, no concrete evidence is available to support this contention, and no study has evaluated the image consistency of discography as influenced by this factor. By observing the consistency of two images in relation to needle tip position we aimed to clarify the importance of needle positioning in discography. One hundred and ninety-two patients (324 discs) receiving steroid intradiscal therapy in whom discography of the same disc was performed twice at a 2-week interval and in whom the needle tip position was within the acceptable range (as defined by us) were studied. The patients were divided into two groups: in group G, in whom the needle tip was within a limited range on both discograms, and group P, in whom the needle tip was in this range on only one discogram. Image consistency was compared roentgenographically in the two groups. The consistent image rate for the total number of discs was 48.5%, being significantly higher in group G (53.2%) than in group P (39.0%). The rates were lower in the nucleus pulposus and the posterior portion of the disc than in the other disc areas, but were significantly higher in group G (85. 4% and 75.0%, respectively, for these two areas). The necessity for accurate needle tip positioning was proved roentgenographically. PMID- 9732559 TI - Which vertebrae should be assessed in diagnosing osteoporosis by plain radiography? Comparative study of radiographic findings and bone mineral density measured by dual energy X-ray absorptiometry. AB - In 1990, the Ministry of Health and Welfare of Japan advocated a criterion for the diagnosis radiographic bone atrophy in lumbar vertebrae. We carried out a comparative study of this criterion (i.e. , radiographic bone atrophy grading) and bone mineral density measured by posteroanterior and lateral dual energy X ray absorptiometry. We investigated the second to fourth lumbar vertebrae of 47 postmenopausal women (mean age, 67.2 years; range, 57-82 years), to determine which vertebra would most accurately indicate osteoporosis. A significant difference in bone mineral density (P < 0.01) was found between each grade of radiographic bone atrophy in all of the second, third, and fourth vertebrae examined. The highest correlation (r = 0.714) was found between the radiographic grades and the bone mineral density on lateral dual energy X-ray absorptiometry of the third lumbar vertebrae. Our visual determinations were dependable, radiographic and a examination of the third lumbar vertebra rather than that of other lumbar vertebrae was more helpful for the detection of radiographic bone atrophy. Plain radiographic film can provide a rough estimation of bone atrophy. PMID- 9732560 TI - Effect of exercise on tibial and lumbar vertebral bone mass in mature osteopenic rats: bone histomorphometry study. AB - The effect of moderate running exercise on tibial and lumbar vertebral bone mass was examined in mature osteopenic rats by bone histomorphometry. Ten 37-week-old female Wistar rats, with bone loss resulting from being fed a relatively low calcium diet for 14 weeks after ovariectomy at the age of 23 weeks, were randomly divided into two groups of five animals each; control and exercise groups. The exercise consisted of treadmill running at 12 m/min for 1 h per day on 5 days per week for 12 weeks. During the exercise period, all animals were fed a standard calcium diet. After 12 weeks of exercise, bone histomorphometry was evaluated for cancellous bone (secondary spongiosa) of the proximal tibia and the fourth lumbar vertebra and for cortical bone of the tibial shaft. The findings suggested that in the mature osteopenic rat, there was a beneficial effect of moderate running exercise with adequate calcium intake on bone mass only in a weight-bearing long bone, the tibia. The mechanism for increased bone mass appeared to be both decreased bone resorption and increased bone formation in cancellous bone and increased bone formation in cortical bone. PMID- 9732561 TI - Influence of ligation of the internal iliac veins on the venous plexuses around the sacrum. AB - Excessive bleeding is a significant problem during total sacrectomy. Ligation of the internal iliac veins to control bleeding from the pelvic venous plexus has been reported to be mandatory. However, despite ligation of the internal iliac veins, excessive hemorrhage from the pelvic and epidural venous plexuses is often encountered. We postulated that ligation of the internal iliac veins increases blood loss during total sacrectomy and we investigated the influence of ligation of the internal iliac veins on the pelvic and epidural venous plexuses in white rabbits. We also investigated the influence of the animal's operative position on the epidural venous pressure. Venography was performed to study the differences in blood flow patterns before and after ligation of the internal iliac veins. Without ligation, contrast medium passed into the inferior vena cava, but not into the epidural venous plexus. The epidural venous plexus was contrast-filled when the internal iliac veins were ligated. The pressure in the internal iliac veins was increased with their ligation, and decreased with ligation of the abdominal aorta. The pressure was also decreased with intentional bleeding from the epidural venous plexus, and with changing the animal's position to headdown. Ligation of the internal iliac veins leads to congestion of the pelvic venous and epidural venous plexuses. Ligation of the internal iliac arteries and positioning the animal headdown were effective ways to resolve the congestion in these venous plexuses. PMID- 9732562 TI - A brief review for orthopedic surgeons: fatigue damage (microdamage) in bone (its determinants and clinical implications). AB - Bone modeling can slowly strengthen bones to keep their strains below bone's microdamage (MDx) threshold. When that condition is satisfied the slow basic multicellular unit (BMU)-based remodeling can usually repair the little MDx that occurs anyway, and some always does. While this arrangement minimizes fatigue fractures of whole bones or trabeculae, they can still happen if: (A) drugs, disease, or dead bone impair MDx repair; (B) if bone loads increase faster than the sluggish modeling can strengthen bone to meet the new loads, and/or faster than remodeling can repair the increased MDx; (C) if a cyst, tumor, or surgery removes enough bone to let strains in the remaining bone reach or exceed the MDx threshold; (D) if abnormal properties of bone as a material cause too much MDx to repair; (E) if altered modeling and remodeling thresholds cause an osteopenia that lets normal activities cause bone strains in or above the MDx threshold range; (F) or if strains in the bone supporting a load-bearing implant reach or exceed bone's MDx threshold. PMID- 9732563 TI - From Wolff's law to the mechanostat: a new "face" of physiology. PMID- 9732564 TI - Osteosarcoma of bone. AB - The definition of osteosarcoma requires that a malignant tumor of bone produce osteoid or bony matrix. With this as a basic definition, osteosarcoma of bone can be divided into several clinicopathologic entities based on clinical, roentgenographic, and pathologic features. The tumors can be broadly divided into those arising within the bone and those arising on the surface of bone. Most intraosseous osteosarcomas are high-grade malignant tumors that occur in children and adolescents. A small number occur in older patients, and they may be related to a preexisting condition, such as Paget's disease or radiation. The site of the lesion has prognostic importance. Osteosarcoma of the jaws is associated with an especially good prognosis, whereas the same kind of tumor involving the skull has a very poor prognosis. Most osteosarcomas of the surface of bone are well or moderately differentiated and are associated with an excellent prognosis. PMID- 9732566 TI - Complaints about nurses: the unlearnt lessons. PMID- 9732565 TI - Nursing must find its raison d'etre. PMID- 9732567 TI - Male catheterization by female nurses: a small-scale survey. AB - A small-scale questionnaire survey of nurses in one Welsh district general hospital examined the views of nurses in relation to catheterization of male patients. The findings demonstrate that while most nurses agree that it is acceptable for females to catheterize male patients, most female nurses do not undertake the procedure as they incorrectly believe there are either local or national policies that prevent patients from being catheterized by nurses of the opposite sex. This often results in patients waiting longer than necessary to be catheterized, and nurses (usually male nurses) from other clinical areas being requested to catheterize a patient for whom they are not caring. Such beliefs and practices stifle the development of knowledge on this issue. PMID- 9732568 TI - Wound assessment: measuring the area of a leg ulcer. AB - As part of an holistic and systematic assessment, the area of a leg ulcer can be a useful measurement for evaluating the wound repair process. The treatment of a chronic leg ulcer can, if not monitored, become a sequence of dressings and bandaging. Thus, a timely recognition of improvement or deterioration of the wound condition may be overlooked. Area change often depends upon the original size and pathology of the ulcer as well as the treatment method. Perhaps the most important factor which can be derived from area measurements is the rate of area change. Measurements taken at regular intervals give a good indication of a wound's healing progress. Computer technology can be used to assist in accurate area measurement. This information gives the clinician vital information on the treatment's efficacy and the data can be used for further statistical or case study analysis. PMID- 9732569 TI - Breast awareness project for women with a learning disability. AB - In 1995/6, while conducting a quality assurance evaluation in a residential group home for adults with a learning disability, the subject of breast screening and the importance of early detection of breast abnormalities was raised by a member of staff. At the time, the Mulberry Trust had a quality standard that women between 50 and 64 years who are supported in continuing care should attend breast screening clinics on a 3-yearly basis if they wish. A pilot scheme was devised based on the premise that breast awareness for service users should be promoted. It was decided that the scheme should include a breast examination, conducted on a monthly basis, ideally by the service users themselves, or by suitably trained staff on their behalf. The pilot scheme was implemented using available research and training was provided for qualified nursing staff within the trust. The training covered breast cancer prevention and breast awareness. After 7 months the scheme was evaluated and changes were made to policy and practice, including consultation with the trust's ethics panel regarding implementation of procedures. Training for staff in breast awareness continues and the scheme is slowly being introduced across the trust to enable all service users to be involved, both in residential homes and community settings. However, in future, the emphasis will be on identifying changes in the breast during normal care routines, such as bathing and dressing, as opposed to formal, clinical examination. PMID- 9732570 TI - The link between Down's syndrome and Alzheimer's disease: 1. AB - This article, the first of two parts, considers the link between Down's syndrome and Alzheimer's disease and how this link has been a significant factor with regards to research into the aetiology of Alzheimer's disease. It describes some of the suggested causes of Alzheimer's disease in people with Down's syndrome. The diagnosis, signs and symptoms of Alzheimer's disease are briefly discussed. The second article concludes with the implications of Alzheimer's disease in people with Down's syndrome for family careers, services and nurses. PMID- 9732571 TI - Caring for patients with morbid obesity in hospital. AB - Caring for the patient with morbid obesity may require adaptation of routine nursing care; however, as with all patients, care should be tailored to individual needs. This article aims to highlight the specific needs of patients with morbid obesity and discusses ways of addressing these needs. The activities of living model is commonly used to assess, plan, implement and evaluate nursing care. This model, used to identify the potential needs of a patient with obesity, will provide the framework for the article. Documentation of procedures used for patient care, e.g. lifting and bathing, must be made in the patient's care plan enabling other nurses caring for the patient to identify quickly the most appropriate and safe way to care for the patient. PMID- 9732572 TI - Radiology nursing: legal issues surrounding the expanded role. AB - The annual conference for the Royal College of Nursing's Forum of Nurses in Radiology and Cardiology took place in Birmingham last year. Participants were asked to consider the ways in which their professional practice could develop within The Scope of Professional Practice (UKCC, 1992a) guidelines and the possible problems they feared could arise. The legal issues which arose from their answers were then analysed. The aim of this article is to examine, in the light of the legal issues, the chosen areas of expansion within the nurse's role and the concerns felt by the participants in order to draw up lessons for nursing generally. PMID- 9732573 TI - Proactive risk management: documentation of patient care. PMID- 9732574 TI - [Promotion of perception in neonatal intensive care]. PMID- 9732575 TI - [Preoperative and postoperative care after correction of transposition of great arteries]. PMID- 9732576 TI - [Medicine between mother and child. II. The newborn in the hospital. Chances and dangers from the experts]. PMID- 9732577 TI - [Is nursing in need of special ethics?]. PMID- 9732578 TI - ["Breathe deeply"--pediatric nursing education, slightly different]. PMID- 9732579 TI - [Sebastian Kneipp's active health promotion]. PMID- 9732580 TI - [Toxic and less toxic plants]. PMID- 9732581 TI - [Requirement for regulated patient documentation. The documentation has to show the complete course of disease]. PMID- 9732582 TI - [Objectively caring--sociopolitical and ethical dimensions in pediatric nursing- 6. Hannover Pediatric Nursing Congress]. PMID- 9732583 TI - [Children and adolescents in future health care--comprehensive health care for children requires effective pediatric departments]. PMID- 9732584 TI - Work makes nurses sick. PMID- 9732585 TI - Value nurses and they will stay. PMID- 9732586 TI - Cost of keeping nurses on holiday island. PMID- 9732587 TI - Home comforts. PMID- 9732588 TI - Understanding dysfunction. PMID- 9732589 TI - Cold feet but warm debate. PMID- 9732590 TI - Testing our intuition. PMID- 9732591 TI - Study notes. PMID- 9732592 TI - Postnatal ills. PMID- 9732594 TI - Clinical equals. PMID- 9732593 TI - Countdown to change. PMID- 9732595 TI - Maintaining networks. PMID- 9732596 TI - Excellence in action--Charter Mark 1998. AB - Last year, 365 Charter Mark awards were made to public sector organisations who deal directly with the public. In this report, we describe how organisations may apply for the award, the benefits of gaining the award and the judging process. PMID- 9732597 TI - Aggression and violence: examining the theories. AB - Theories of aggressive and violent behaviour fall into a confusing range of categories. In this review, the author attempts to make sense of the different concepts and describes the theory that underpins each of them. PMID- 9732598 TI - Issues in infertility nursing: broadening the debate. AB - The lack of dialogue between nurses and feminists around fertility and infertility treatments detracts from the development of assisted conception nursing. In this article, the author examines the existing literature and argues for a stronger nursing contribution to this controversial subject. PMID- 9732599 TI - Medical admissions units: the role of the nurse practitioner. AB - In this article, the author describes the development of a medical admissions unit. He also highlights the introduction of an advanced nurse practitioner role and the contribution this has made to raising the quality of patient care. PMID- 9732600 TI - Managing the menopause. AB - This article discusses the effects of the menopause and ways in which nurses can advise women on help and treatments that are available to help allay their symptoms. This article should be read in conjunction with other CE articles and these are referred to at the appropriate points. PMID- 9732601 TI - Home help. PMID- 9732602 TI - Tide is turning on inequality of care. PMID- 9732603 TI - Study matters. PMID- 9732604 TI - Children beware. PMID- 9732605 TI - Far too little and too late. PMID- 9732607 TI - Time to go. PMID- 9732606 TI - Personal touch. PMID- 9732608 TI - Conflict of interests. PMID- 9732609 TI - Working where the clients are. PMID- 9732610 TI - When two souls meet. AB - As a nurse specialist working in a regional gender identity unit, Rosemary Grimshaw is frequently called on for advice. Here she describes her role and some common misconceptions about transsexualism. PMID- 9732611 TI - Transmission of HIV from mother to child. AB - Over 4,000 cases of women with human immunodeficiency virus (HIV) have been reported from every area of the UK (PHLS 1997). There are over 300 births each year to HIV-positive mothers, with the highest prevalence in London. This joint report finds that HIV-positive women are generally not offered routine testing and treatment, and recommends that a programme to prevent the transmission of HIV from mother to baby is needed urgently. PMID- 9732612 TI - Psychological management of stoma-related concerns. AB - In this article, the author discusses how nurses can identify, prevent and treat the psychological consequences of stoma surgery for patients and their families. PMID- 9732614 TI - Paying the price. PMID- 9732613 TI - Promoting health in young people. AB - This article discusses aspects of health that might affect young people and ways in which nurses can promote healthy living and support this group of clients during a potential period of anxiety. PMID- 9732615 TI - Tailor made for midwives. PMID- 9732616 TI - Post-registration education and practice (PREP) PMID- 9732617 TI - Anthea does the rounds. PMID- 9732618 TI - No smoke without fire. PMID- 9732619 TI - Moving too fast. PMID- 9732620 TI - Dream on. PMID- 9732621 TI - Video nation. PMID- 9732622 TI - Joint effort. PMID- 9732623 TI - Sexual rights. PMID- 9732624 TI - Is it time to cut corners? PMID- 9732625 TI - Watchful eye. PMID- 9732626 TI - A helping hand. PMID- 9732627 TI - A class act. PMID- 9732628 TI - War against the deadly sting. PMID- 9732629 TI - Time to catch up. PMID- 9732630 TI - Adverse reactions to food. AB - A significant number of people believe that they are 'allergic' to certain foods. Here, in an edited account of a recent report from the National Dairy Council (1998), the causes and management of food intolerance are discussed. PMID- 9732631 TI - Raising the profile of nursing: how to organise a study day. AB - This article provides practical advice of organising a study day. It is based on the authors' experience of organising a successful event in their own hospital. PMID- 9732632 TI - A comparison of two depot injection techniques. AB - In the study reported in this article, the researchers attempted to raise awareness among practitioners of the importance of intramuscular drug administration technique in reducing injection site complications following antipsychotic depot injections. They also aimed to improve and expand the scope of present practice by comparing the effect of two accepted techniques, the 'air bubble' and 'Z-track' on these complications, and demonstrate that the air bubbles technique is more effective in reducing seepage and causes less discomfort. A 'within subjects' design was used, and Likert scales for scoring subjective and objective assessment of complications were established and scored at each injection. The study showed that there was no significant difference between the effects of either technique. PMID- 9732633 TI - Abuse of power in the nurse-client relationship. AB - A small number of health professionals are at risk of stepping over the boundaries of acceptable behaviour towards their clients. While sexual misconduct is clearly defined, the author argues that other inappropriate behaviours are harder to define--especially in nursing where touch is an important component of care. PMID- 9732635 TI - New areas in which nurses could take on work traditionally done by doctors. PMID- 9732634 TI - Hyperlipidaemia. AB - This article discusses the etiology of hyperlipidemia and methods of diagnosis and treatment, associating this with the risk of coronary heart disease, so that nurses will develop their understanding and advise patients appropriately. PMID- 9732636 TI - Milestones on the road to equal pay. PMID- 9732638 TI - The price of stress. PMID- 9732637 TI - Learning the script. PMID- 9732639 TI - How to save eight lives a year. PMID- 9732640 TI - Health promotion for everyone. PMID- 9732641 TI - We never close. PMID- 9732642 TI - 50 years of NHS nursing. PMID- 9732643 TI - The RCN's enduring relationship. PMID- 9732644 TI - Computer care. PMID- 9732645 TI - Research: getting it funded. AB - In this summary of the ENB's recent guidelines on preparing research proposals, we review the strategic purpose of the research funding agencies and the process of submitting a successful research proposal. PMID- 9732646 TI - From research to practice: a review of the literature. AB - In this article, the author considers whether nursing research is more likely to be implemented in a climate which recognises the importance of research evidence. She then examines the capacity of research to improve the quality of nursing care. PMID- 9732647 TI - Pressure damage prevention: basing practice on evidence. AB - As part of an initiative to develop evidence-based practice at the Northern General Hospital, Sheffield, a three-part project was undertaken. The aims were to identify barriers to using research in nursing, establish a baseline of nurses' knowledge and its influence on their practice in one essential area of nursing care--pressure damage prevention--and develop a strategy for change which took account of the findings from the first two parts of the project. In this article, the authors describe the second part of the project which examined nursing knowledge and practice with reference to the management of pressure damage prevention. The findings are discussed and the authors recommend that nurses integrate into their practice evidence from sources such as systematic reviews. PMID- 9732648 TI - Radiotherapy nursing: understanding the nurse's role. AB - Radiotherapy nurses are integral members of the multidisciplinary team. In this article, the author describes the role and makes recommendations for its future development. PMID- 9732650 TI - The menopause: sexually-related problems. AB - This article discusses the physiological and psychosocial effects of the menopause on women and relates these effects to sexual relationships so that nurses are better informed when advising clients/patients. PMID- 9732649 TI - Nurses' access to library and information services. AB - The author reports on the impact of recent changes in the funding and organisation of library and information services which support the education of nurses. In order that nurses might better contribute to evidence-based practice and clinical effectiveness initiatives, the author calls for greater communication and co-operation between library and health services. PMID- 9732651 TI - Communication in general practice: recognition and treatment of mental illness. AB - From previous studies there is a lot of evidence that in primary care settings, many patients tend to express their mental problems in terms of physical symptoms. Therefore, the general practitioner (GP) needs to recognize mental problems at an early stage. Early recognition allows for adequate treatment that might speed up recovery. The present article reports on a study exploring the GP's ability to recognize mental illness, the communication style that is supposed to support this ability, the subsequent treatment of mental problems, and the patient's recovery. Two databases were used. First, an observation study, involving 351 videotaped consultations held by 15 GPs, yielded information on communication style and recognition abilities. Patients in this study were selected randomly. The second database obtained treatment data and measures of patient recovery from a 1-year follow-up study dealing with the treatment and course of mental illness. Patients in this study were selected because their GPs considered their problems "mainly psychosocial by nature". Half of them were categorized within psychological and social diagnostic categories of the International Classification for Primary Care (ICPC), the other half were categorized within physical disease categories, with an assessment by the GP that the complaints were mainly psychosocial. Results showed no significant relationships between the recognition of mental illness and nine communication features supposed to induce these abilities. There was a tendency however, for a positive association between recommended communicative behaviour of the GP and his or her tendency to give frequently psychosocial evaluations of the patient's complaints. Also, there was a negative tendency between this recommended behaviour and the degree of agreement between the GP's evaluation and the score on a psychiatric screening questionnaire. This agreement is called "accuracy". Frequent psychosocial evaluations were related to exploring behaviour and mental health referral in case of psychosocial complaints. Further, relationships between the GPs' recognition ability and various measures of patients' recovery did not prove univocal. Both positive, negative and absent relationships were found. PMID- 9732652 TI - Asthma education programme in Russia: educating patients. AB - To address the recent rise in asthma morbidity and mortality in Russia, an intervention study was conducted to improve asthma diagnosis, treatment and prevention. US recommendations for asthma management were adapted for use in educating Moscow families with children with asthma. Two hundred and fifty-two children with asthma aged 4-14 years receiving health care in eight Moscow public health clinics together with their parents were enrolled in the study to see whether US teaching manuals for asthma management would be acceptable and effective in Russia. Children at four of the clinics with recent asthma attacks were randomly assigned to either the education or control group to test if patient education and guided asthma care would improve outcomes for patients. Modern medications were made available to both groups to see if training in the US guidelines was necessary to get physicians to use the medications. Children with recent asthma attacks at the other four clinics were defined as comparison group 1 to control for the possible effect of medication availability. All children at the eight clinics who had no asthma attacks composed comparison group 2 to see if the outcomes for these children would change over time. One-year follow-up results showed significant improvement in asthma self-management skills of children and parents, in terms of asthma treatment, only among those in the education group. Significant increases were observed in the subgroup of children in the education group using anti-inflammatory drugs for asthma control. Children in the education group had markedly increased peak flow rates and reduced daily peak flow variability as compared to control and comparison groups. There was a significantly greater reduction in doctor visits by the education group of children compared to control. Presumably, changes in parents' and children's behaviour in terms of asthma treatment and prevention skills, proper treatment of the disease and access to medications could be responsible for reducing asthma morbidity in children. PMID- 9732653 TI - Information booklets about cancer: factors influencing patient satisfaction and utilization. AB - Providing patients with adequate information is an important component of care. This exploratory study investigated factors influencing patient satisfaction with and utilization of information booklets. The research was conducted in two stages. In stage 1, five commonly used cancer information booklets were reviewed by 36 Australian patients who were either receiving chemotherapy or had just completed treatment. Data were collected on patient satisfaction with, preference for and utilization of information booklets. In addition data were collected on variables identified in the literature as potentially influencing patient satisfaction, including patient characteristics, presentation and readability of booklets, and the timing of provision. A high level of satisfaction was found for all five information booklets, although a clear preference for one particular booklet emerged. The most notable feature of this booklet was its readability level (grade 8); in contrast the other booklets were written at levels equivalent to grades 11-12. Stage 2 focused on the side effects of patient information preference style on their satisfaction and recall of information presented in two booklets in the course of their treatment. No differences were found between patients who seek information and those who avoid it. The findings of this study suggest that patients' information needs may be better met if information booklets are written in plain English, and presented to patients prior to treatment. Future studies incorporating a larger sample of patients and greater selection and variety of information booklets are required to further determine if patient characteristics and features of booklet presentation influence patient satisfaction and preference. PMID- 9732654 TI - Sex differences in illness beliefs and illness behavior in patients with suspected coronary artery disease. AB - The aim of this study was to explore sex differences in illness beliefs and behavior in patients with suspected coronary artery disease (CAD). Twenty-eight patients, 16 women and 12 men, were interviewed. The results show that both men and women think of CAD as a 'men's disease' and have equal knowledge of CAD risk factors. However, especially the men considered their own risk of developing CAD lower than their estimated probability of their own sex and as low as their estimated risk for women. Both men and women did not attribute their symptoms indicative of CAD to their heart. Women, especially those who did not attribute their symptoms to their heart, had a longer patient delay than men, although their symptoms were indicative of CAD. To conclude, men as well as women should be made more aware of their own risk of developing CAD and of the manifestation of CAD symptoms. Physicians could be encouraged to ask patients more explicitly and thoroughly about their illness beliefs, to check their knowledge and inform them about CAD. PMID- 9732655 TI - Journal writing as a social support strategy for parents of premature infants: a pilot study. AB - BACKGROUND: Having a premature baby is acknowledged to be stressful to parents. Journal writing combines practical, emotional and informational support that may be useful to these parents. METHODS: We conducted a study to assess the potential for promoting journal-writing for parents receiving social support in a special care nursery (SCN). Parents were provided with educational material on journal writing, and subsequently surveyed concerning their journal-writing during their child's hospitalization. RESULTS: Of the 73 parents enrolled, 32% kept a journal; of these, 73% felt it helped considerably in reducing stress, and 68% used it as a means of addressing the most stressful elements of their nursery experience. Journals were used primarily to document involvement in care (45%), record keeping (36%), and organization of thoughts (27%). All of those who kept a journal recommended it for use by other parents. CONCLUSIONS: Encouraging parents to keep a journal is a constructive way of dealing with the SCN-related stress. PMID- 9732656 TI - Problem-posing in education: transformation of the practice of the health professional. AB - This study was developed by a group of professionals from different areas (nurses and educators) concerned with health education. It proposes the use of a problem posing model for the transformation of professional practice. The concept and functions of the model and their relationships with the educative practice of health professionals are discussed. The model of problem-posing education is presented (compared to traditional, "banking" education), and four innovative experiences of teaching-learning are reported based on this model. These experiences, carried out in areas of environmental and occupational health and patient education have shown the applicability of the problem-posing model to the practice of the health professional, allowing transformation. PMID- 9732657 TI - Emotions, coping and the need for support in families of children with cancer: a model for psychosocial care. AB - In the case of childhood cancer, the personal threats are severe for both the child, the parents and other family members. For the child, there is the threat to physical integrity, safety, security, and above all, to life. For the parents, there is the threat of losing the child. However, a number of studies have shown that psychopathological disturbances are rarely found in children with cancer or their parents. We may conclude from this that most children and parents use coping strategies that protect them from developing psychopathology. In organising support for families with a child with cancer, much can be learned from children's and parent's perceptions and reactions. When problems of adjustment arise, a thorough analysis of how children and parents perceive their situation, as well as an extensive analysis of their coping efforts, is necessary to direct effective supportive actions. A psychosocial support model is proposed which can be helpful in interpreting these emotions and coping strategies. PMID- 9732658 TI - The assessment of pain in the cognitively impaired elderly: a literature review. PMID- 9732659 TI - Goal attainment and life satisfaction: variables under study. PMID- 9732660 TI - Niagara Geriatric Mental Health Outreach Team: private partnerships in care. PMID- 9732661 TI - St. Joseph's Villa Community Bathing Program: nursing innovation in action. PMID- 9732662 TI - Nursing assistants reduce aggressive behaviour during bathing cognitively impaired nursing home residents. PMID- 9732663 TI - Learn to love your GP. PMID- 9732664 TI - Amniotomy to shorten spontaneous labour. PMID- 9732665 TI - No win, no fee or no fault? PMID- 9732666 TI - Damage to the pelvic floor: causes, prevention and treatment. PMID- 9732667 TI - Iron supplementation in pregnancy: research-based practice? PMID- 9732668 TI - Maternal mental health. 3: An action plan for midwives. PMID- 9732669 TI - Making it happen. Organizing an overseas study trip. PMID- 9732670 TI - The quiet midwife. PMID- 9732671 TI - Aquanatal training course. PMID- 9732672 TI - My clients and other animals. PMID- 9732673 TI - [Usefulness of a nursing dictionary]. PMID- 9732674 TI - [To explain pain and make it understood: forms and resources of explanatory discussions]. PMID- 9732675 TI - [Pain and nursing care]. PMID- 9732676 TI - [Insecurity ... the effect of the evolution of a concept on the development of nursing knowledge]. PMID- 9732677 TI - [The homeless in emergency services: their contacts with nurses]. AB - This research paper is based on the homeless, as representatives of extreme poverty but also as clients of public hospitals: how far does the rising number of homeless people in the hospital emergency departments pose a problem to the nurses? An epidemiological survey carried out in the emergency departments of Toulouse-Purpan, goes in search of the features of the homeless people. The problematics of this essay is situated at the interface of the three elements dealt with in the original questioning: the homeless, the nurses and the University Hospital. The hypothesis is the following: "The rising number of homeless people welcomed in the emergency departments of the university hospitals is revealing of the tensions to which the nurses are subjected, torn as they are between the specificity of the request, the assignments of the emergency departments and the professional values and skills". Tensions to which the nurses are subjected are identified thanks to the analysis of their speech, on the basis of their perceptions and the image they have of the homeless. These perceptions are interpreted as portent of certain types of practices: dominated by rejection, denial, extreme rationality, questioning, motherly practices. PMID- 9732678 TI - [Acute postoperative pain in the adult: effect of its conception on the method of care]. PMID- 9732680 TI - [Revista Rol de Enfermeria completes 20 years] [In Process Citation] PMID- 9732681 TI - [Nursing and communication]. PMID- 9732679 TI - [Preventing the pain and discomfort of premature infants: a new method of registration of the EEG signal]. PMID- 9732682 TI - [20 Years of dietetics] [In Process Citation] PMID- 9732683 TI - [The future of nursing care in Europe]. PMID- 9732684 TI - [Smoking and diabetes. An intervention protocol]. AB - Tobacco smoking habit has been considered like an important cause of diabetes mellitus complications. Health education efficiency has been supported by many experiences in order to promote tobacco withdrawal by personal counselling with a direct nurse role at primary health care level. This study was carried out by all this information with tre straight goal of describing changes in the diabetic tobacco behaviour after a stepped counselling and a primary care nurse following. Description details are given of the smoking diabetic population counselled and listed at the same time in a nurse control clinic over a 18 months period. A Smokers Helping Scheme (SHS) was used. SHS understood that tobacco withdrawal is behavioural changing process with a few steps. Smokers were given with special written material for thes purpose. Results data were caught by the researchers from the chronical patient's census and from the tobacco program control sheets. Tobacco withdrawal was verified by espirated air CO determination (Smokorlizar system) and has been maintained by the 25% of managed diabetic people in this investigation. We strongly believe that nursing diabetic tobacco counselling by SHS acts like a behavioural modificator as results are showing. Health state improvement and a better life quality have been got on the diabetic people that has been managed. PMID- 9732686 TI - [Radial and ulnar arteries]. PMID- 9732685 TI - [Reflections on Martha E. Rogers' theory]. AB - This article is a brief review on the assertions made by Martha E. Rogers. If one enlarges these assertions, one arrives at the conclusion that each individual lives his/her own designated life, totally different from other people; hence, having a unique living experience. A whole set reality about human nature becomes known by considering an energy field, as well as subatomic particles, to be the fundamental unit of life. As Ms. Rogers see it, "man lives in a probabilistic universe". This affirmation drives us to the limits of uncertainty since we can never really learn all the health-nursing procedures. PMID- 9732687 TI - [Image of the nursing professionals in hospital emergency units]. PMID- 9732688 TI - [System of design and organization of protocols]. PMID- 9732690 TI - [Vitamin and mineral requirements in women III]. PMID- 9732689 TI - [Accidental destruction of a colostomy. Case report]. AB - The case which we describe in the following article is an example of the application of new advances in stomatology as well as an example of perfect coordination among medical professionals. A basic element for high quality health care. We describe the accidental removal of a loop colostomy with detachment and colaspe of the stoma and the corresponding treatment followed in order to maintain the viability of the stoma. Finally, we mention the advantages obtained in choosing a conservative treatment for this kind of complicated stomas. PMID- 9732691 TI - [Prinicipal vitamins and minerals and their importance]. PMID- 9732692 TI - [Epidemiology of pressure ulcers or the danger of a new Tower of Babel]. AB - Pressure epidemiology is a fundamental facet in meeting this important challenge to the health care system. The studies done on epidemiology signify an important effort by their authors, although their validity may be conditioned by methodological and conceptual aspects. The proliferation of studies about pressure epidemiology in our country, some of which having methodological conceptions not in agreement with the existing bibliography about this topic in other countries, make it necessary to gain a grasp of the basic epidemiologic aspects applied to the study of pressures as a health problem. This paper presents a bibliographical review on the subject and states proposals adapted to the characteristics of our immediate environs. This paper includes a review of these concepts: prevalence, incidence, recurrence, and severity; in addition to the most utilized variables in describing the pressure problem. Different strategies for the elaboration of epidemiological studies about pressures are also analyzed. Furthermore, the minimum information which authors of said studies need to facilitate in their reports are mentioned. PMID- 9732693 TI - [Tracheostomy cannulas]. AB - Tracheostomic clyster pipes are mechanisms which allow one to artificially maintain the permeability of the air passageway under those circumstances when it is not possible to breathe by the conventional way. Nowadays, there is a wide variety of clyster pipes available to medical professionals. Each of these has its own traits and precise applications. This article describes those clyster pipes most frequently used in hospitals and outpatient clinics; furthermore, their use, care and therapeutic applications are mentioned. PMID- 9732694 TI - [Living with chronic renal insufficiency]. AB - In Spain today there are some 27,000 patients suffering from chronical renal insufficiency to such an extent that they require treatment which artificially performs their kidney functions or have had a kidney transplant. The basic artificial renal function therapies are hemodialysis and peritoneal dialysis. Both therapies, which are usually well-tolerated although not exempt from complications, allow patients to have an acceptable rehabilitation and quality of lifestyle. Theoretically, a kidney transplant is the ideal solution; however, the lack of donor organs, organ rejection and patients; particular circumstances place limits on this therapeutic option. Nevertheless, whatever therapy is chosen, patients must make adaptions in their life style to include following a diet, administering prescribed drugs, applying specific hygienic practices, and the like. All of these require the patient to have an appropriate educational support structure. PMID- 9732695 TI - LY294002-mediated inhibition of phosphatidylinositol 3-kinase activity triggers growth inhibition and apoptosis in CD40-triggered Ramos-Burkitt lymphoma B cells. AB - Cells of the Epstein-Barr virus genome-negative Ramos-Burkitt lymphoma (Ramos-BL) B cell line can be rescued from antigen receptor (AgR)-triggered growth inhibition and apoptosis by signals transduced through their surface CD40. This study investigates whether phosphatidylinositol 3-kinase (PI3-kinase), which has been reported to be intimately involved in the regulation of normal and neoplastic cell growth, plays a role in CD40-promoted Ramos-BL B cell survival and uses the selective and reversible PI3-kinase inhibitor, LY294002 (LY). LY mediated inhibition of PI3-kinase activity triggers growth inhibition and leads to the processing of caspase-3, caspase-3-like activity, cleavage of the death substrate poly(ADP-ribose) polymerase (PARP), and apoptosis from the G1 phase of cell cycle. These data indicate that constitutive PI3-kinase activity is critical for Ramos-BL B cell progression through the cell cycle such that if this PI3 kinase-dependent pathway(s) is inhibited, the cells default to apoptosis. Signals transduced through CD40 abrogate LY-triggered caspase-3-like activity and PARP cleavage but fail to inhibit LY-triggered growth inhibition, processing of caspase-3, and apoptosis. Likewise, in the presence of LY, signals transduced through CD40 abrogate AgR-triggered caspase-3-like activity and PARP cleavage but fail to inhibit AgR-triggered growth inhibition, caspase-3 processing, and apoptosis. The LY-mediated induction of growth inhibition and apoptosis occurs in the presence of the CD40-induced anti-apoptotic protein Bcl-XL. Taken together these data indicate that the CD40 of Ramos BL B cells is linked to PI3-kinase independent and -dependent routes of survival: CD40-mediated inhibition of AgR triggered caspase-3-like activity, PARP cleavage, and CD40-triggered Bcl-XL expression are PI3-kinase-independent, whereas PI3-kinase is critical for CD40 mediated rescue of this cellular population from AgR-triggered growth inhibition, caspase-3 processing, and apoptosis. PMID- 9732696 TI - Host resistance against Listeria monocytogenes is reciprocal during the course of infection in alymphoplastic aly mutant mice. AB - The aly is a unique spontaneous autosomal recessive mutation in mice that causes a systemic defect of lymph nodes and Peyer's patches. We investigated host resistance against Listeria monocytogenes infection in the mutant. The 50% lethal dose of L. monocytogenes in aly/aly mice was 10-fold higher than their heterozygotes, termed aly/+mice, or their wild-type C57BL/6 mice. The bacterial growth in the spleens and livers of aly/aly mice was more efficient early in infection, and their listericidal activity of peritoneal macrophages was higher than those of aly/+mice. In contrast, the complete elimination of bacteria from the spleens and livers of infected mice in the late stage of infection, in which a T-cell-dependent mechanism is required, was delayed in the aly/aly mice. Moreover, an acquired resistance against secondary infection with L. monocytogenes was markedly diminished in the aly/aly mice. The production of endogenous interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF alpha), which are critical in antilisterial resistance, was reduced in the aly/aly mice during the infection. The production of IFN-gamma, and interleukin-4 was also diminished in the spleen cell cultures of aly/aly mice when stimulated with heat-killed L. monocytogenes or the T-cell receptors were directly stimulated with anti-CD3-epsilon monoclonal antibody. These results suggest that acquired immunity against L. monocytogenes infection is attenuated in aly/aly mice, and that the insufficient production of IFN-gamma and TNF-alpha might be involved in the immunodeficiency. PMID- 9732697 TI - Noninvolvement of IL-4 and IL-10 in tolerance induction to experimental autoimmune thyroiditis. AB - The mechanisms of tolerance induced with deaggregated mouse thyroglobulin (dMTg) in experimental autoimmune thyroiditis (EAT) is not yet well defined. As shown previously, the induction and maintenance of tolerance require CD4+ T cells exerting active regulatory function to prevent EAT induction. To examine whether Th2 cells are responsible for resistance we injected anti-IL-4 and anti-IL-10, separately or together, into CBA (H2k) mice at the time of MTg pretreatment to study the role of IL-4 and IL-10 in tolerance induction. Our results show that tolerance can be well established without involving IL-4 or IL-10. To determine whether IL-4 was involved in tolerance induction in another EAT-susceptible strain, IL-4 knockout mice on B10.Q background were similarly pretreated with dMTg and immunized. These IL-4 knockout mice exhibited very good tolerance. The lack of response to EAT induction was not due to IL-4 deficiency, since immunized IL-4 knock-out control mice developed severe EAT. Moreover, resistance was strong in IL-4 knock-out mice also given anti-IL-10. The data in both susceptible strains show that IL-4 and IL-10 play a small role in induced resistance to EAT. PMID- 9732698 TI - Identification of a human CD8+ T lymphocyte neo-epitope created by a ras codon 12 mutation which is restricted by the HLA-A2 allele. AB - Point mutations in the ras proto-oncogenes, notably at codon 12, are found in high frequency of human malignancies and, thus, may be appropriate targets for the induction of tumor-specific T cell responses in cancer immunotherapy. In this study, we examined the mutant ras protein sequence reflecting the substitution of Gly to Val at position 12 as a putative point-mutated determinant for potential induction of an HLA-A2-reactive, CD8+ cytotoxic T lymphocyte (CTL) response. We identified the ras 4-12(Val12) sequence as a minimal 9-mer peptide, which displayed specific binding to HLA-A2 by T2 bioassays. Peptide binding to HLA-A2 on T2 cells was weak and required coincubation with exogenous beta(2) microglobulin to facilitate and enhance complex formation. In contrast, the wild type ras 4-12(Gly12) peptide failed to bind to HLA-A2 even in the presence of beta(2)-microglobulin, consistent with the hypothesis that the point mutation creates a C-terminus anchor residue. A CD8+ CTL line against the ras 4-12(Val12) peptide was derived in vitro from a normal HLA-A2+ donor using a model culture system consisting of T2 cells as antigen presenting cells pulsed with exogenous mutant ras peptide and beta(2)-microglobulin plus cytokines (interleukin-2 and 12). Functional characterization of CD8+ CTL line revealed (1) peptide-specific and HLA-A2-restricted cytotoxicity against a panel of peptide-pulsed targets; (2) no specific lysis using the normal ras peptide sequence; (3) half-maximal lysis with exogenous peptide of approximately 0.3 microM; (4) lysis of HLA-A2+ B cell lines infected with a recombinant vaccinia virus construct encoding the point mutated human K-ras gene; and (5) specific lysis of the HLA-A2+ SW480 colon carcinoma cell line expressing the naturally occurring K-ras Val12 mutation. Maximal lysis of SW480 cells occurred following interferon (IFN)-gamma pretreatment, which correlated with enhanced HLA-A2 and ICAM-1 (CD54) expression. Specificity of lysis was revealed by the absence of lysis against a HLA-A2+ melanoma cell line (+/- IFN-gamma), which lacked the mutant Val12 mutation, and the inability of an irrelevant CD8+ CTL line to lyse SW480 (+/- IFN-gamma) unless the appropriate exogenous peptide was added. These findings demonstrated that tumor cells may endogenously process and express mutant ras epitopes, such as the 4-12(Val12) sequence, albeit in limiting amounts that may be potentiated by IFN gamma treatment. These data support the biological relevance of this sequence and, thus, may have important implications for the generation of ras oncogene specific CTL responses in clinical situations. PMID- 9732699 TI - Developmental changes and functional properties of human memory T cell subpopulations defined by CD60 expression. AB - The present study was undertaken to examine developmental changes of T cells expressing CD60 and their functional properties. Three-color immunofluorescence analysis revealed that the CD60 antigen was preferentially expressed on a proportion of memory (CD45RO+) CD4+ T cells, but less on memory CD8+ T cells, while this antigen is undetectable in naive (CD45RO-) T cells. A frequency of memory CD4+ T cells expressing CD60 in the peripheral blood was negligible in newborns and gradually increased with advancing age. CD60+ memory CD4+ T cells showed stronger proliferative responses to PPD and produced higher levels of IL-4 and IL-10 than CD60- ones, whereas production of IL-2 and IFN-gamma was similarly found in both cell subpopulations. In addition, it was shown that efficient helper activity for Ig production by B cells was predominated in CD60+ memory CD4+ T cells. These results suggest that CD60 may be primarily expressed on the functionally differentiated memory effector cells among circulating CD45RO+ CD4+ T cells. PMID- 9732700 TI - Induction of endogenous mammary tumor virus in lymphocytes infected with murine acquired immunodeficiency syndrome virus. AB - Mice infected with murine acquired immunodeficiency syndrome (MAIDS) virus developed lymphoadenopathy and profound immunodeficiency. Concomitantly the expression of endogenous mammary tumor virus (MTV) mRNA increased significantly, especially for the 1.7-kb 3' open reading frame (ORF) mRNA encoding MTV superantigen. B cell lines that are established from MAIDS mice and exhibit superantigen activity also express a high level of 1.7-kb endogenous MTV and mRNA. Infection of a B cell tumor line in vitro with retrovirus containing the cloned MAIDS virus gene induced superantigen activity and this cell line also expressed the 1.7-kb superantigen coding MTV 3' ORF mRNA. These results strongly suggest a link between MAIDS virus infection and the induction of endogenous superantigen activity. This may play an important role in the pathogenesis of the MAIDS virus. PMID- 9732701 TI - Impaired negative selection in CD28-deficient mice. AB - T cell antigen receptors (TCR) expressed on developing T cells can react with self-peptides presented by proteins encoded by the major histocompatibility complex (MHC). Depending on the relative strength of these interactions, thymocytes are either negatively selected as potentially autoreactive and deleted or positively selected to become mature T cells. Developmental selection may also be regulated by signals in addition to those mediated through the TCR. In peripheral T cells, the CD28 receptor plays an important role in enhancing the survival and expansion of T cells activated by TCR engagement. Therefore, we have investigated the role of CD28 in regulating the selection of thymocytes using CD28-deficient mice. Surprisingly, we found a 50% increase in cell number in the thymi of CD28-deficient compared to wildtype mice, suggesting that CD28 might play a role in negative selection. Negative selection of double-positive thymocytes was found to be significantly reduced in response to either antigen or antibody crosslinking of the TCR complex in CD28-deficient animals. This was not due to a generalized defect in thymocyte survival as thymocytes from CD28 deficient and wildtype mice displayed similar sensitivity to apoptosis initiated by either gamma-irradiation or dexamethasone. In contrast to its role in T cell activation and survival in the peripheral immune system, the CD28 receptor appears to participate in the intracellular signaling events that result in negative selection in the thymus. PMID- 9732702 TI - Effect of a monoclonal antibody against interleukin-4 on suppression of antigen induced arthritis in mice by oral administration of the inducing antigen. AB - We investigated a role for interleukin-4 (IL-4) in suppression of T-cell-mediated antigen-induced arthritis (AIA) in mice by oral administration of the inducing antigen. For this investigation, a monoclonal antibody (11B11 mAb) that specifically neutralizes IL-4 was employed. AIA was induced by immunization with methylated bovine serum albumin (mBSA) (day 0) followed by intraarticular injection of mBSA into the ankle joint. To induce oral tolerance, mBSA was administered orally once a day from day-5 to-1. The 11B11 mAB was injected i.p.. 30 min before each oral administration of mBSA. Oral administration of mBSA resulted in marked suppression of AIA. Suppression of the joint inflammation of the oral antigen was significantly diminished by treatment with 11B11 mAb. The mAb treatment was also followed by blockade of suppression of proliferative responses of lymphoid cells to mBSA by oral antigen. Furthermore, secretion of IL 4 was significantly increased following oral administration of mBSA and the increased IL-4 secretion was markedly reduced by treatment with 11B11 mAb. There was a decrease in production of IFN-gamma in orally tolerized mice that was blocked by the IL-4-neutralizing mAb. Thus, treatment with an anti-IL-4-mAb appears to be effective in blocking suppression of AIA by oral administration of the inducing antigen. The results also suggest that IL-4 may play a role in down regulation of T-cell-mediated inflammation by feeding pathogenic antigens. PMID- 9732703 TI - Protein I/II of oral viridans streptococci increases expression of adhesion molecules on endothelial cells and promotes transendothelial migration of neutrophils in vitro. AB - As accumulation of leukocytes in perivascular tissues is a key step in inflammatory disorders, we have analyzed in the present work the up-regulation of expression of adhesion molecules, such as E-selectin, ICAM-1, and VCAM-1, on human endothelial cells, in response to protein I/II, a modulin from Streptococcus mutans OMZ 175. Using cultured human saphenous vein endothelial cells (HSVEC), we demonstrated that protein I/II directly and specifically up regulated E-selectin, ICAM-1, and VCAM-1 expression. We confirmed also that the up-regulation of adhesion molecules in HSVEC is mediated by lectin activity for NANA- and fucose-containing receptors. The ability of protein I/II to promote the transendothelial migration of neutrophils was then examined. Using Transwell inserts, we found that protein I/II, in promoting the up-regulation of adhesion molecule expression, stimulates neutrophil migration through endothelial cells. These events may play a role in the etiology of inflammatory responses leading to the various pathologies associated with oral viridans streptococci. PMID- 9732704 TI - The glycosylphosphatidylinositol-anchored form and the transmembrane form of CD58 are released from the cell surface upon antibody binding. AB - The adhesion molecule CD58 is expressed on the cell surface in both a transmembrane form and a glycosylphosphatidylinositol (GPI)-anchored form. Here we report that CD58 is released from JY cells following cross-linking by immobilized anti-CD58 monoclonal antibodies. Antibodies to other cell surface proteins, as well as PMA and LPS, did not trigger CD58 release. The release resulted from membrane cleavage, since biotin-labeled CD58 was released from biotinylated cells, and down-modulation of CD58 surface expression accompanied accumulation of soluble CD58 IN culture media. We have previously reported the isolation of JY variant cells, which lack expression of GPI anchored proteins and thus express only the transmembrane form of CD58. Here we show that these variant cells release CD58 upon crosslinking, indicating that the transmembrane isoform is released, probably by proteolysis. Antibodies directed to the cytoplasmic domain of CD58, in contrast to antibodies against an extracellular epitope of CD58, did not react with released CD58, supporting a membrane cleavage mechanism. It is also shown that CD58, released from [3H]ethanolamine-labeled JY cells, contained ethanolamine. This result demonstrated that the GPI-anchored CD58 can be released in parallel to the transmembrane isoform and that this release does not result from proteolytic cleavage, since cleavage by a protease would have removed the ethanolamine. The present data suggest that the two isoforms of CD58 are released upon antibody binding and that their release is mediated by distinct mechanisms. PMID- 9732705 TI - Bring back the rotating internship. PMID- 9732706 TI - Diagnosis of chest discomfort simplified. PMID- 9732707 TI - Abortion in proportion. PMID- 9732708 TI - The best treatment may be no treatment at all. PMID- 9732709 TI - Reporting of gender-related information in clinical trials of drug therapy for myocardial infarction. AB - BACKGROUND: Concern has been expressed that women are not adequately represented in clinical trials evaluating treatments for medical conditions they commonly experience. This study was designed to assess the reporting of data on women in recently published trials of drug therapy for myocardial infarction, including those funded by an agency with a gender-related policy. METHODS: All randomized controlled trials and meta-analyses of drug therapies for myocardial infarction published in The New England Journal of Medicine, The Lancet, The Journal of the American Medical Association, the Annals of Internal Medicine and the British Medical Journal from January 1992 to December 1996 were evaluated. On preliminary review 102 articles met the inclusion criteria; these were reviewed in detail, and 59 were excluded. Two reviewers independently extracted gender-related information from the 43 articles; discrepancies were resolved by consensus. RESULTS: Women presented up to 48% of the trial participants (mean 24.1%). In the trials funded by an agency with a gender-related policy, only 16.8% of participants, on average, were women. Of the 43 articles in the sample, only 14 (32%) provided gender-related results. Funding from an agency with gender-related policy did not affect the reporting of gender-related information. Subgroup analyses were provided for 14 (32%) of the 43 trials, including 2 (29%) of 7 trials funded by an agency with a gender-related policy. Of the 12 trials that included interaction analyses (excluding the 2 trials in which secondary analyses were conducted specifically to identify differences between women and men), 7 (58%) conducted an interaction analysis to determine if women responded differently than men; for one of these the interaction analysis was for a secondary outcome measure (drug safety). Only 5 (12%) of the 43 articles mentioned the differences between men and women in the Discussion section; 2 of these were studies that used secondary analyses to examine sex differences. Of the 5, only 1 was funded by an agency with a gender-related policy. INTERPRETATION: Women were poorly represented in the randomized controlled trials in this sample, regardless of whether the trials were funded by an agency with a gender-related policy. Structured reporting of gender-related information for clinical trials may improve the quality of information available about women and therefore facilitate the application of research findings to the care of women. PMID- 9732711 TI - The challenge of diversity in the delivery of women's health care. PMID- 9732710 TI - Survivors of sexual abuse: clinical, lifestyle and reproductive consequences. AB - BACKGROUND: In recent years, an increase in the prevalence of sexual abuse of women has been reported in Canada and elsewhere. However, there are few empirical data on the extent of the problem in Canadian aboriginal populations. The authors investigated the presence of a reported history of sexual abuse and other health determinants in a sample of women attending a community health centre with a substantial aboriginal population. This allowed determination of whether reported sexual abuse and its associated demographic and health-related effects were different for aboriginal and non-aboriginal women. METHODS: A sample of 1696 women was selected from women attending a community health centre in a predominantly low-income inner-city area of Winnipeg for a cross-sectional survey designed to study the association between sexual behavior and cervical infections. The survey was conducted between November 1992 and March 1995 and involved a clinical examination, laboratory tests and an interviewer-administered questionnaire. A substudy was conducted among 1003 women who were asked 2 questions about sexual abuse. RESULTS: The overall response rate for the main study was 87%. Of the 1003 women who were asked the questions about sexual abuse, 843 (84.0%) responded. Among the respondents, 368 (43.6%) were aboriginal. Overall, 308 (36.5%) of the respondents reported having been sexually abused, 74.0% of the incidents having occurred during childhood. The prevalence was higher among aboriginal women than among non-aboriginal women (44.8% v. 30.1%, p < 0.001). Women who had been sexually abused were younger when they first had sexual intercourse, they had multiple partners, and they had a history of sexually transmitted diseases. In addition, non-aboriginal women who had been sexually abused were more likely than those who had not been abused to have been separated or divorced, unemployed and multiparous and to have used an intrauterine device rather than oral contraceptives. Aboriginal women who had been sexually abused were more likely than those who had not been abused to have been separated or divorced, unemployed and multiparous and to have used an intrauterine device rather than oral contraceptives. Aboriginal women who had been sexually abused were more likely than those who had not been abused to have had abnormal Papanicolaou smears. The proportion of smokers was higher among the abused women than among the non-abused women in both ethnic groups. INTERPRETATION: A history of sexual abuse was associated with other clinical, lifestyle and reproductive factors. This suggests that sexual abuse may be associated with subsequent health behaviors, beyond specific physical and psychosocial disorders. Aboriginal and non-aboriginal women who have suffered sexual abuse showed substantial differences in their subsequent health and health related behaviours. PMID- 9732712 TI - Goin' to the country: challenges for women's health care in rural Canada. PMID- 9732713 TI - Understanding women's health through data development and data linkage: implications for research and policy. PMID- 9732714 TI - Lessons in women's health: body image and pulmonary disease. PMID- 9732715 TI - Women who use injection drugs: the social context of risk. PMID- 9732716 TI - Sexual health of women with disabilities. PMID- 9732717 TI - Disability and childbirth: meeting the challenges. PMID- 9732718 TI - Not all your patients are straight. PMID- 9732719 TI - Abreast in a boat--a race against breast cancer. PMID- 9732720 TI - Not alone: peer support through audio teleconferencing for rural women with breast cancer. PMID- 9732721 TI - A pain consultation clinic for women. PMID- 9732722 TI - Healthy living for immigrant women: a health education community outreach program. PMID- 9732723 TI - Providing primary health care to immigrants and refugees: the North Hamilton experience. PMID- 9732724 TI - The poor are different from you and me. PMID- 9732725 TI - Canadians participate as unique US project puts death under the microscope. AB - The Death in America Project, sponsored by an American philanthropist, is designed to find inadequacies in knowledge about the course and outcomes of care of dying patients. The participating scholars include 3 Canadian physicians. PMID- 9732726 TI - What women don't know could kill them. PMID- 9732727 TI - [Nootropic drug evaluation for general practice]. AB - Treatment with "nootropic drugs" of patients suffering from dementia is often described as arbitrary. To define the potential usefulness of nootropic drugs, effects and side effects, economic aspects in comparison to other treatment approaches were studied. A summary of literature published concerning these criteria underlines the efficacy of nootropics in improving the symptoms at the beginning and in postponing the progress of the disease. The majority of substances does not lead to severe side effects. There are not enough studies comparing the effects of the substances or on prophylactic effects. Delayed admission to care units leads to a positive cost effect of nootropics. Thus, also for economic reasons it seems advisable to treat patients with nootropics. Better effects are gained when nootropic treatment is combined with training and changing of the structure of the environment. A proposal for a rational treatment with nootropics is derived from the data. PMID- 9732728 TI - [Psychiatric evaluation in accordance with the transsexualism regulation. An experience-based plea for guidelines and against arbitrary bias]. AB - Today patients want to change their Christian names not so much for social support but in order to obtain sex change operations. For this reason among others we caution against casual and careless evaluations and plead for conscientiousness and thoroughness. The spectrum of 16 evaluation cases in the year 1995 is presented. Following this and looking for guiding principles, the most important aspects are discussed: the dilatable frame of the written law, the examiner's diagnostic process, the duty of being well informed, reconstruction of development as the first goal of examination, real-life self-testing and therapy, the obligation to report one's own findings, change of Christian name and indication of operation, evaluation for gender changing, problems of the judge expert relationship and of future development. PMID- 9732729 TI - [Transsexualism. Current status of research and clinical practice]. AB - In the past decades the cultural and social status of transsexuals has changed considerably. They have established their own organizations, obtained legal rights and access to health insurance services. The professional point of view has changed as well considerably. Very different personality structures, developmental courses and sexual preferences are described. As differential diagnoses homosexual and transvestitic developments, psychoses, early-onset personality disorders, adolescence crises, culturally induced gender dysphorias, and intersexual disorders have primarily to be considered. In therapy, the old confrontation psychotherapy vs. surgery has widely been relinquished. Patient's age is nowadays much lower. Sex ratio in Germany is approaching for the first time 1:1. So far worldwide a preponderance of biologically male transsexuals is reported. The total number of adult transsexuals in Germany is estimated 2000 4000. PMID- 9732730 TI - [The Chabot case. Assisted suicide from the psychiatric viewpoint]. AB - The international discussion on physician-assisted dying as well as the recent development in North Australia and Oregon point to a growing tendency to favour assisted suicide as against killing on request--last not least for reasons of public acceptance. The decision of the Supreme Court of the Netherlands in a case of suicide assisted by a psychiatrist gives the opportunity to discuss the problem from the psychiatric point of view. PMID- 9732731 TI - [Violence in partnership of immigrant women from Turkey]. AB - Forty-one female immigrants from Turkey, who were victims of violence in their relationships and diagnosed with "reaction to severe stress and adjustment disorders" (F43, ICD-10) were compared with a controle-group of the same origin, sex, age and diagnosis who had not suffered from such violence. The frequency of emergency treatment and suicide attempts (in the last month before the first consultation) of the victims of violence were higher than in the controle-group. The victims of violence less frequently live in large families and were more often employed than the controle-group. The duration of time until the immigration of the partner was significantly higher in the group of victims than in the controle-group. The results are discussed with regard to studies concerning other ethnic groups and specific aspects of psychodynamics and migration. PMID- 9732732 TI - [Compulsory treatment in psychiatry from the viewpoint of the patient. A prospective study]. AB - During a 6-month period all patients who had been treated involuntarily in a state-funded mental mental health unit were studied prospectively. They were interviewed after the first compulsory measure and at the end of treatment. A control group of patients (not experiencing involuntary treatment) were interviewed as well. The aim of the study was to establish whether the groups differed with respect to judgement of subjective well-being and satisfaction with treatment. The stability of these judgements over time and especially changes in perception and acceptance of involuntary treatment during the course of treatment were also monitored. the 36 patients in the study group perceived themselves as being far less ill at both time points and were less satisfied with treatment than the 29 control patients. At the end of treatment, all patients judged themselves to be significantly more ill retrospectively and patients of the study group were more satisfied with treatment than at the beginning. When questioned in the days after an involuntary measure, 25% of the patients concerned denied that any measure had taken place against their will; at the end of the hospital stay this figure had increased to 42%. In contrast, four control patients complained that they had been involuntarily subjected to treatment. In discordance with the previously published literature, the acceptance of involuntary treatment declined over time. PMID- 9732733 TI - [Defenses in rating scales and therapy success]. AB - Evaluation of psychotherapeutic or psychopharmacological treatment frequently uses rating scales without regard for patient's defensiveness. This study demonstrates on 309 psychotherapy patients treated with inpatient client-centered therapy and followed-up 1 year after treatment that defensiveness, measured with validity scales of personality inventories MMPI-K, FPI-R and Giessen-Test, influences ratings (scales CGI, HAMA, HAMD, BRMES) before and after therapy. Patients who deny psychopathology on admission do so at discharge and to some extent at follow-up. The extent of effect is significantly diminished with an increase in frankness in these patients, indicated by the significant improvement in validity scales. Inpatient client-centered psychotherapy improved defensiveness in the case of initial self-criticism and diminished it in the case of initial retentiveness. PMID- 9732734 TI - [Assisted suicide after discharge from the psychiatric clinic]. AB - Assisted suicide has become a topic often and controversally discussed in international literature. Psychiatry and mentally ill, however, have been neglected, probably because the question of competence of judgement and will are a major issue in this matter. Two cases are reported, one of a 60-year old man and a women of 87 years, who have committed suicide a few days after having been dismissed from stationary psychiatric care. The reason of admission had been a psychotic event with manifest AIDS-disease in the first, the public utterance of her wish to die with support of, the swiss "Help to Die" organization "Exit", in the latter. Ethics and means of preentive interventions in such cases are discussed. PMID- 9732735 TI - [Assisted suicide in dementia. Ethical considerations between paternalism and autonomy]. AB - This article discusses the problem of euthanasia presenting the case of an 82 year-old man with progressive dementia. Difficulties encountered during daily clinical work are described and analysed, in order to clarify decisions on ethical, legal and professional medical grounds. General decisions concerning life-sustaining measures may be qualitatively improved if the situation of the individual is constantly assessed and considered within the treatment process. PMID- 9732736 TI - [Immunoglobulin therapy in Gilles de la Tourette syndrome]. AB - It has known for a long time that Sydenham's chorea and tics, as seen in Gilles de la Tourette's syndrome (GTS), are phenomenologically very similar. Tics may occur as symptoms of acute Sydenham's chorea or persist over years as residual symptoms. Investigating of children suffering from GTS, including obsessive compulsive symptoms, have provided signs of a poststreptococcal autoimmune process but also shown that treatment based on immunological interventions has been effective. We treated a 14-year-old boy showing all diagnostic criteria of GTS, familial susceptibility, and an increase in the antibody titer of streptococcal antigens with 75 immunoglobulins i.v. over 5 days. Response to this therapy was good regarding motor tics, vocal tics, and behavioral symptoms such as disturbed impulse control which still persisted after 9 months. These findings and the successful therapy underline reports of the literature and point to a pathogenetic mechanism of an immunologically triggered disturbance of the striatal dopaminergic system, at least in a subgroup of GTS. PMID- 9732737 TI - [Change in gender identity in male pseudohermaphroditism. Differential diagnosis of transsexualism and intersexuality]. AB - Gender identity change in intersexual patients is rare. We report on the transsexual development of a patient with male pseudohermaphroditism. The case report emphasizes the importance of a thorough differential diagnosis. PMID- 9732738 TI - [Report on development of standards in treatment and assessment of transsexual patients]. PMID- 9732740 TI - ["Chronic fatigue syndrome"]. PMID- 9732739 TI - [Entactogenic drugs "ecstasy" (MDMA), "eve" (MDE) and other ring-substituted methamphetamine derivatives. A new class of substances among illegal designer drugs?]. PMID- 9732741 TI - ["Opiate positive" in urine drug screening test after eating poppy seed cake]. PMID- 9732742 TI - [Historical and current construction principles for psychiatric clinics]. PMID- 9732743 TI - [Supplementary remarks]. PMID- 9732744 TI - [Commentary. DGPPN recommendation for basic psychiatric documentation]. PMID- 9732745 TI - Lack of retrovirus gene expression in teratocarcinoma stem cells is limited to the nucleus. AB - Experiments are described showing that the block to retrovirus replication by undifferentiated embryonal carcinoma cells can be overcome by fusion with differentiated cell karyoplasts but not cytoplasts. This supports previous notions that the expression of input proviral genes in teratocarcinoma stem cells is under different controls than in differentiated cells and indicates that the exercising of this control is limited to the nucleus of the embryonal carcinoma cell. PMID- 9732746 TI - Transcriptional control of the expression of mouse globin genes in myeloma x erythroleukemia cell hybrids. AB - Fusions were made between thymidine kinase deficient (TK-) Friend Cells inducible for hemoglobin production, and immunoglobulin-producing, hypoxanthine-guanine phosphoribosyltransferase-deficient (HGPRT-) myeloma cells. Hybrids were selected in hypoxanthine-aminopterin-thymidine (HAT) and identified by isozyme analysis and chromosome counts. All hybrids resembled the myeloma cell line in mode of growth and were immunoglobulin secretors. All hybrids did not express hemoglobin and were uninducible for hemoglobin production with dimethyl sulfoxide (DMSO). Hybridization of genomic globin DNA probes with hybrid-derived nuclear and cytoplasmic mRNAs blotted to nitrocellulose filter indicated that lack of expression of the globin genes in the hybrids was due to lack of transcription. PMID- 9732748 TI - A temperature-sensitive DNA synthesis mutant isolated from the Chinese hamster ovary cell line. AB - A temperature-sensitive DNA synthesis mutant, tsC8, was isolated from mutagenized Chinese hamster ovary cells by the fluorodeoxyuridine suicide technique. The tsC8 cells showed inhibition of DNA synthesis at the nonpermissive temperature (NPT) with little effect on initial levels of RNA and protein synthesis. Temperature arrested tsC8 cells had G1 or S DNA content and the temperature-sensitive (ts) period of the tsC8 cell cycle was the interval between the G1/S border and the middle of the S period. The tsC8 cells were unable to enter the S phase when exposed to the NPT during the G1 period of the cell cycle. When S phase tsC8 cells were shifted to the NPT, they incorporated [3H]thymidine at rates similar to the parental cell type for only 2 h, indicating a ts defect in DNA synthesis. The tsC8 mutation is expressed in a recessive manner and is in a gene distinct from those affected in other DNA synthesis mammalian cell mutants. PMID- 9732747 TI - Regulation of human globin gene expression in mouse erythroleukemia x human fibroblast hybrid cells. AB - A somatic cell hybrid, XX-8, was obtained from a fusion of tetraploid mouse erythroleukemia cells with human Lesch-Nyhan skin fibroblasts. This hybrid cell was previously shown (1) to produce human beta- but no human gamma-globin mRNA sequences after induction with dimethylsulfoxide. In this study we show that: (a) human beta- and gamma-globin genes are present in XX-8 cells in approximately equal numbers; (b) no human gamma-globin mRNA sequences can be detected in either the cytoplasmic or nuclear RNA fractions even with several different inducers; (c) after induction the human beta-globin gene is converted from a DNase I insensitive or closed structure to a DNase I open configuration, while the human gamma-globin gene remains closed; and (d) no human beta-globin polypeptide can be detected in the intact induced cells, indicating that fibroblast globin genes, even when induced to make mRNA in an erythroid environment, do not synthesize an RNA that is translated efficiently. PMID- 9732749 TI - Expression of human transferrin receptor is controlled by a gene on chromosome 3: assignment using species specificity of a monoclonal antibody. AB - The monoclonal antibody OKT-9 has been shown to recognize the human transferrin receptor. We have exploited the species specificity of OKT-9 to map a gene controlling human transferrin receptor expression to chromosome 3, using human mouse somatic cell hybrids. The gene for the human transferrin receptor and the gene controlling transferrin expression may be linked in humans. PMID- 9732750 TI - Bromodeoxyuridine resistance in CHO cells occurs in three discrete steps. AB - Four independent mutants were isolated from mutagenized cultures of CHO cells by sib selection on the basis of resistance to a low concentration (2.6 x 10(-5) M) of BrdU. All four lines were stable, but all had about 100% of the wild-type (WT) specific activity of thymidine kinase (TK). None of the four yielded derivatives resistant to a high level of BrdU (2 x 10(-4) M) in one step even after mutagenesis, but variants resistant to 4-6 x 10(-5) M BrdU could be isolated at frequencies of about 2 x 10(-5)/cell. At frequencies of 10(-4)-10(-5), the second step mutants gave colonies resistant to 2 x 10(-4) M BrdU. The second and third steps of resistance were correlated with partial and complete reduction, respectively, in the specific activity of TK, suggesting that the variants may be genotypically heterozygous and homozygous-negative at the tk locus. The first step of BrdU resistance was dominant and appeared to result from a mutation in the gene from ribonucleotide reductase, since in vitro assays on partially purified preparations showed that the reductase activity in mutant cells was less sensitive to BrdUTP than in WT cells. PMID- 9732751 TI - Assignment of an oligomycin-resistance locus to human chromosome 10. AB - An oligomycin-resistant variant of human fibrosarcoma HT1080 was isolated and characterized as nuclear and codominant. The mutant was stable, was not cross resistant to respiratory inhibitors, and it contained a mitochondrial ATPase which was less sensitive to oligomycin. Hybrids formed between the human mutant and a mouse cell line expressed the resistance phenotype. By a detailed karyotypic analysis of these hybrids using trypsin-Giemsa banding it was found that resistance to oligomycin correlated with the retention of two human chromosomes 10. The hybrid lines contained only mouse mitochondrial DNA as shown by analyses of mitochondrially synthesized proteins and mitochondrial DNA. The study assigns an ATPase oligomycin-resistance locus to human chromosome 10 and suggests that mouse and human subunits can combine in a functional enzyme complex. PMID- 9732752 TI - Linkage of the leuS, emtB, and chr genes on chromosome 5 in humans and expression of human genes encoding protein synthetic components in human--Chinese hamster hybrids. AB - We isolated interspecific hybrids between normal human leukocytes and a Chinese hamster ovary cell line that has mutations in three genes, leuS, emtB, and chr, all of which are linked to chromosome 2. The conditionally lethal mutation in the leuS gene in this cell line affects leucyl-tRNA synthetase and renders the cell line nonviable at 39 degrees C. The mutation in the emtB locus alters ribosomal protein S14 and results in the cell line being resistant to the protein synthesis inhibitor, emetine, while the mutation in the chr locus renders the cells resistant to sodium chromate. The interspecific hybrids were selected at 39 degrees C so that they were required to retain and express the human leuS gene. Ten out of ten such heat-resistant hybrids also expressed the human emtB and chr genes. Segregants selected as having lost the human emtB gene simultaneously lost the human chr and leuS genes as well. The linkage relationship between these three genes has thus been conserved during the evolution of the human and Chinese hamster genomes. All three genes were localized to human chromosome 5. Furthermore, our results indicate that the ribosomal protein product of the human emtB gene is incorporated into functional ribosomes in place of the human corresponding Chinese hamster protein, raising several interesting questions concerning the coordinate regulation of genes encoding ribosomal proteins in mammalian cells. PMID- 9732754 TI - [What is the future of Tzanck's cytodiagnosis?]. PMID- 9732753 TI - Derepression of genes on the human inactive X chromosome: evidence for differences in locus-specific rates of derepression and rates of transfer of active and inactive genes after DNA-mediated transformation. AB - Mouse-human hybrid cells that contained an inactive human X chromosome were treated with agents known to alter gene expression and to perturb DNA methylation. 5-Azacytidine greatly increased the rate of derepression of HPRT on the inactive X, while butyrate and dimethyl sulfoxide had smaller effects. Ethionine did not change the rate of derepression. Derepression of two other X chromosomal loci, PGK and GPD, was also detected. The rate of derepression of PGK was 20-fold higher than the rate for HPRT. Derepression events at the two loci appeared to be independent. Hybrids expressing derepressed X-chromosomal genes had more variable levels of human enzyme activities when compared to control hybrids. HPRT+ clones did not appear after transfer of purified DNA from a cell hybrid containing an inactive human X into HPRT- recipients, but such clones did appear after transfer of DNA from derivative cells in which HPRT had been derepressed. PMID- 9732755 TI - [Antiproteases in every shape]. PMID- 9732756 TI - [Contact vasculitis caused by topical agents with non-steroidal anti-inflammatory agents or analgesics]. AB - INTRODUCTION: Topical non steroidal antiinflammatory drugs (NSAID) are recently used in France. Seven cases of contact vasculitis due to topical NSAID are reported. PATIENTS AND METHODS: The clinical and histological features and follow up data of seven patients with contact vasculitis due to topical NSAID were retrospectively reviewed. RESULTS: The mean age of the seven patients (four women, three men) was 39 years. The topical NSAID used were: ketoprofene in four cases, mephenesine in one case and phenylbutazone in two cases. Cutaneous lesions occurred after a mean time of four days. Histological examination of a skin biopsy specimen showed a leucocytoclastic vasculitis in two cases, a lymphocytic vasculitis in two cases and a mixed vasculitis in three cases. Previous sensibilization to the drug was noted in five cases. Cutaneous patch tests with the drugs were positive in all cases. CONCLUSION: The risk of systemic reaction after oral ingestion of the culprit drug may be considered in these patients. PMID- 9732757 TI - [Peri-sudoral lipoma]. AB - INTRODUCTION: We identified in our files 11 cases of a new variant of the superficial lipoma recently described by Hitchcock, Hurt and Santa Cruz, characterized by the presence of eccrine sweat glands and denominated "adenolipoma of the skin". PATIENTS AND METHOD: We examined 1,742 skin lesions registered in the Laboratory for Skin Histopathology from January 1989 to August 1996. These lesions were 397 lipomas, 1,325 skin tags (acrochordons) of which 120 with a fatty stroma and 20 connective tissue hamartomas of which 7 superficial lipomatous hamartomas. Among these lesions we looked for those corresponding to the prime description of the cutaneous adenolipoma. RESULTS: We identified 11 cases of adenolipoma, i.e. a frequency of 0.65 p. 100 for the whole examined lesions and 2 p.100 among the 524 lesions with a fatty component (lipomas, lipomatous hamartomas and fibrolipomas). The mean age of the patients was 50 years and the sex-ratio F/M 1.75. The lesions were localized on the lower limbs (7 cases), especially on the thighs (4 cases), on the trunk (3 cases) and on the shoulder (1 case). DISCUSSION: The adenolipoma is quite different from the common cutaneous lipomas. It develops within the dermis or the fatty layer. It is a solitary lipoma most often localized on the proximal parts of the limbs, especially on the thighs. Histologically it presents as a lobulated and capsulated tumor where eccrine sweat glands and ducts are present inside the fatty lobules. Apparently the adenolipoma originates from the fat pad around the sweat coils without proliferation of the sweat glands or ducts; this entity should therefore be denominated "perisudoral lipoma" rather than adenolipoma. It has to be differentiated from the skin tags with a fatty stroma (fibrolipomas) and from the superficial lipomatous hamartomas. PMID- 9732758 TI - [Rapid diagnosis of cutaneous leishmaniasis and histoplasmosis by direct microscopic tests]. AB - BACKGROUND: Cutaneous leishmaniasis and disseminated histoplasmosis can be diagnosed by direct examination of pathology specimens after Giemsa staining. CASE REPORT: An HIV-infected man developed cutaneous leishmaniasis and disseminated histoplasmosis with buccal lesions concomitantly. Smears stained with the RAL 555 kit provided the diagnosis of these two diseases. DISCUSSION: This case illustrates how direct microscopic examinations can provide inexpensive, rapid and safe diagnosis of infectious diseases. Direct microscopic examinations are routine practice in a tropical dermatology unit. PMID- 9732759 TI - [Kasabach-Merritt syndrome of the leg associated with osteolysis or Gorham sign]. AB - INTRODUCTION: Kasabach-Merritt syndrome and Gorham's sign are two uncommon and severe, sometimes life-threatening, complications in infants with vascular lesions. Their association has been described in rare cases. CASE REPORT: An infant with a vast congenital angiomatous lesion including an extensive lymphatic component, developed active regional osteolysis then suddenly suffered disseminated intravascular coagulation of the leg. Medical treatment was unsatisfactory. After unsuccessful use of low molecular weight heparin, pentoxifyllin and alpha interferon, amputation of the leg was required to avoid a fatal outcome. DISCUSSION: Kasabach-Merritt syndrome does not develop on classic immature hemangiomas, despite some contradictory statements in the literature. In our case, a complex tumor developed in association with a lymphatic malformation. The association of Kasabach-Merritt syndrome with osteolysis (Gorham's sign) does not appear to be fortuitous. Therapeutic management of these severe complications is difficult and requires case by case analysis. PMID- 9732760 TI - [Cutaneous adenolipoma]. AB - INTRODUCTION: Adenolipoma of the skin is an unusual microscopic variant of the solitary lipoma, superposable to the adenolipoma of the breast, characterized by the presence of normal eccrine sweat glands inside the fat proliferation. OBSERVATIONS: Two new cases are reported. DISCUSSION: Adenolipoma of the skin is a benign lesion whose clinical features are similar to those of solitary lipoma. Probably, it is only a histological curiosity in which the eccrine glands are entrapped and carried by the adipose proliferation. PMID- 9732761 TI - [Spindle-cell hemangioendothelioma with monomelic and multifocal form in a child]. AB - BACKGROUND: Spindle-cell hemangioendothelioma is a soft tissue skin tumor recently identified histologically. It can occur at all ages but generally is seen in young adults. The lesion usually occurs as a subcutaneous mass involving the limbs. CASE REPORT: A particular case of spindle-cell hemangioendothelioma was observed in an 18-month-old child. The lesions progressed with a monomelic distribution on the upper limb. Histological diagnosis of spindle-cell hemangioendothelioma was achieved at the age of 6 years. DISCUSSION: The age of the patient and the monomelic distribution is particular in this case of spindle cell hemangioendothelioma, inciting a nosological discussion on this disease and other vascular tumors of childhood and the relationship of these types of lesions with Maffucci's syndrome. Although no anomalies have been detected to date, radiological surveillance is needed as cases of Maffucci's syndrome associated with spindle-cell hemangioendothelioma is described in the literature. PMID- 9732762 TI - [The concept of neutrophilic disease]. PMID- 9732763 TI - [Physician-patient relations in dermatology]. PMID- 9732764 TI - [A case for diagnosis: multiple syringoma]. PMID- 9732765 TI - [Accidental exposure to blood]. PMID- 9732767 TI - [Question of the month: how do you treat frostbite?]. PMID- 9732766 TI - [Dermatology and psychotropic drugs]. PMID- 9732768 TI - [Target-controlled intravenous anesthesia. Who pilots what?]. PMID- 9732769 TI - [Use of Glasgow coma score in prehospital assessment]. PMID- 9732770 TI - [Assessment of a new light guide (Trachlight) for tracheal intubation]. AB - OBJECTIVE: To assess the learning curve of a new lighwand device, Trachlight (Laerdal), for blind orotracheal intubation in patients without foreseen difficulty in airway management. STUDY DESIGN: Open, prospective, clinical study. USERS: Twelve persons practicing anaesthesia (specialists, trainees, nurses) underwent videotape learning and manikin training with ten successful intubation manoeuvres required with the device. METHODS: Each person had to carry out a tracheal intubation in ten consecutive patients undergoing scheduled surgery and without history or clinical sign of difficults airway management. RESULTS: One hundred and twenty patients were included. The overall success rate with the Trachlight was 87%. An easy learning curve was obtained as demonstrated by the low failure rate in the first three patients, and by the success rate on the first or second attempt in the last four patients. There was no significant difference in failure rate with or without muscle relaxation (10 vs 20%, NS). Finally, all failures with the Trachlight were followed by successful intubation using direct laryngoscopy, and no traumatic complications were recorded with the device. CONCLUSION: Trachlight is a new lightwand device enabling blind tracheal intubation with a easy learning curve in patients without difficulty in airway management, even for non-selected operators. PMID- 9732771 TI - [Malignant hyperthermia and appendicular sepsis. Can they be differentiated during surgical procedure?]. AB - OBJECTIVE: To assess the possibility to differentiate clinically intraoperative malignant hyperthermia (MH) and sepsis. STUDY DESIGN: Comparative retrospective study of clinical cases. PATIENTS: Sixteen patients operated on for acute appendicitis and developing clinical signs of MH confirmed or not by in vitro caffeine halothane contracture tests (IVCT). METHOD: To isolate the patients' characteristics with regard to the diagnosis of sepsis and MH crisis. To compare both groups of clinical features with results of IVCT. RESULTS: The diagnosis of MH sensitivity has been excluded in ten hyperthermic patients and confirmed in four others with IVCT. No correlation was existing between the importance of perioperative sepsis, MH features and IVTC results. CONCLUSIONS: This study confirmed the difficulty to differentiate clinically MH and sepsis during surgery. Considering the severe outcome of MH crisis, it is recommended to start the specific therapy even in case of appendicular sepsis. PMID- 9732772 TI - [Use of Glasgow coma scale by anesthesia and intensive care internists in brain injured patients]. AB - OBJECTIVE: To evaluate the quality and reliability of the Glasgow coma scale (GCS) score when determined, in head trauma patients, by trainees in anaesthesiology. STUDY DESIGN: Prospective survey. USERS: One hundred trainees in their first to fourth year of training in anaesthesiology. METHODOLOGY: A questionnaire completed by the trainees concerning: demographic data; place, time and qualification of the physician determining the first GCS score; time and qualification of the physician determining the subsequent GCS score; assessment of the GCS score in case of asymmetrical motor response, tracheal intubation, bilateral eyelid oedema, or circulatory or ventilatory failure. RESULTS: Sixty questionnaires were available for analysis. Lack of compliance with the rules for the GCS score evaluation resulted in many errors by most of the trainees. Only a few of them determined an accurate GCS score in cases of asymmetric motor response or impossibility to determine verbal or ocular response. Finally, GCS scores were determined later only very rarely. CONCLUSION: In order to provide optimal care and allow an accurate assessment of therapeutic efficiency, special attention should be given to the teaching of the GCS scoring method in head trauma patients. PMID- 9732773 TI - [Relations between anesthetists and general practitioners or pediatricians. Results of a cross sectional study among 2281 physicians with private practice]. AB - OBJECTIVE: To assess the relationship of anaesthetiologists (Anaes) with general practitioners (GP) and paediatricians (Paed), possessors of the medical files of children scheduled to undergo ambulatory anaesthesia and participating in postoperative surveillance. STUDY DESIGN: Cross-sectional prospective survey. PERSONS: GP and Paed treating children admitted in September 1994 at the paediatric clinic of the University hospital of Tours for ambulatory surgery. METHOD: Questionnaire including 11 items circulated to 2,181 GP and 100 Paed of the centre of France. RESULTS AND DISCUSSION: Replies were obtained from 1,053 GP (48%) and 64 Paed (64%). The experience of Anaesth in paediatric anaesthesia was the least important criterion for the choice of the clinic (for 67% of Paed and 44% of GP). The rate of contacts of Anaesth with GP and Paed was low (23% of GP and 38% of Paed). Informations concerning anaesthesia (26% of GP and 42% of Paed), or its complications (8% of GP and 8% of Paed) were scarce. The rules of ambulatory anaesthesia were better known by Paed (56%) than by GP (34%), unlike those for autotransfusion (45% of GP and 23% of Paed). A majority of GP (82%) and Paed (73%) would appreciate an anaesthetic report. Most of GP (86%) and Paed (83%) were in favour of more informations, mainly through continuing medical education, on these problems. PMID- 9732774 TI - [Target-controlled intravenous anesthesia]. AB - Target-controlled infusion (TCI) is a new delivery system for i.v. anaesthetic agents with which the anaesthetist targets a plasma drug concentration to achieve a predetermined effect. With this system, the tedious task of calculating the amount of administered drug required to achieve the target concentration is left in charge of a microprocessor which commands the infusion device. TCI has long been used only by a few research teams, but this year a much wider field opens to this delivery system through marketing of Diprifusor, a TCI system specifically designed for administration of propofol in everyday practice. This article describes the rationale for administering i.v. agents through TCI delivery systems, the pharmacokinetic basis of TCI, the regulations and a broad overview of clinical applications, both recent and yet to come. PMID- 9732775 TI - [Different types of nerve injuries in locoregional anesthesia]. AB - Neural damage is a possible complication of central nerve blockade and regional anaesthesia. Damage may be caused by ischaemic, mechanical or chemical mechanisms, which may occur either alone or in combination. Neural ischaemia may be caused by 1) prolonged and severe arterial hypotension, which compromises blood supply to the cord, 2) a spinal haematoma whose main etiological factor is a coagulation abnormality and 3) an intraneural injection. Mechanical trauma by the needle bevel is an important factor of neuropathy, particularly when searching for paraesthesiae. Neurological complications may also result from a direct neurotoxic effect of local anaesthetic agents which is concentration and dose-dependent. Better understanding of these mechanisms will permit the establishment of reliable bases for new preventive strategies. PMID- 9732776 TI - [Hemodynamic effects of hypertonic saline solutions]. AB - Haemodynamic effects of hypertonic saline solutions (HSS) have been extensively studied in animals and humans. Hypertonic sodium chloride (7.5%, 2,500 mOsm.L-1) either alone or combined with colloids, remains the standard solution. The haemodynamic response of HSS observed during treatment of hypovolaemic shock is explained by 1) an increase in preload due to the expansion of the plasma volume and a musculocutaneous vasoconstriction and 2) a decrease in systemic vascular resistance and afterload. A myocardial stimulation has been shown in various experimental conditions and in humans. However, the clinical relevance of this inotropic effect is questionable. Haemorrhagic shock is the main indication for small volume resuscitation with HSS. Other potential situations for the use of HSS are volume replacement in perioperative period, septic shock or burn injury and cardiopulmonary resuscitation. Before recommending the clinical use of HSS, additional clinical studies are required to substantiate the benefits of HSS over colloids. PMID- 9732777 TI - [Anaphylactic shock caused by fibrin glue]. AB - We observed a case of anaphylactic shock in a 68-year-old woman after a nasal intramucosal injection of fibrin glue for telangiectasies therapy. The tests showed an allergy to aprotinin contained in the glue. In the previous years, glue and aprotinin had been administered to the patient several times for nasal bleeding. PMID- 9732778 TI - [A low dose of nalbuphine reverses respiratory depression but not analgesia induced by intraspinal morphine]. AB - Postoperative pain management after scoliosis surgery is based in our institution on intrathecal morphine administration. This case report describes an immediate and major postoperative respiratory depression that occurred in the recovery room, requiring the maintenance of the endotracheal tube. This respiratory depression was reversed by i.v. administration of a low dose of nalbuphine, which allowed tracheal extubation without suppression of morphine-induced analgesia. PMID- 9732779 TI - [Hemolysis can conceal another hemolysis]. AB - We report a delayed haemolytic reaction during the course of a severe HELLP syndrome, which required red blood cell transfusions. Haemolysis was mainly ascribed to the erythrocyte alloantibody anti-Fy a. The fact that this antibody was undetectable before transfusions (despite cross-matching and anti-erythrocyte antibody screening test) emphasizes the limits of these tests for low antibody concentrations. Moreover, this patient, who had a remarkable obstetrical history, received red blood cell transfusions matched for Rhesus and Kell antigens. The opportunity to perform a more extended phenotype screening for patients at high risk to develop multiple alloantibodies is discussed. PMID- 9732780 TI - [Severe form of familial migraine with coma]. AB - The authors report the case of a 43-year-old man experiencing a migraine attack with coma requiring a treatment in an intensive care unit for 9 days. The probable triggering cause as a cerebral arteriography. PMID- 9732781 TI - [Desflurane and errors of gas selection on vaporizer analyzer]. AB - With monochromatic infrared gas spectrometers (MIS), the displayed concentration is computed from measured IR absorption and a gain factor specific for the selected volatile agent (VA). As MIS cannot detect which VA is actually present, the displayed concentration can be very different from the actual one. As bottles and vaporizers are very specific for desflurane, it is impossible to misfill a vaporizer; however an erroneous selection of VA on MIS remains possible. The aim of this study was to assess the displayed concentrations after erroneous vapour selection on the monitor. When either desflurane, or isoflurane or enflurane were delivered at constant concentrations, all VA measured by the MIS, namely desflurane, sevoflurane, isoflurane, enflurane and halothane were successively selected and the displayed concentrations compared with the actual vapour concentration using a Capnomac Ultima (Datex) monitor. Consequences of erroneous selection can be included in three categories: 1) dangerous error, when a displayed concentration is much lower than the actual one, e.g. desflurane or sevoflurane erroneously selected; 2) evident error, when displayed concentration is much higher than 10 vol%; 3) uncomfortable situation, when displayed and actual concentrations are similar, e.g. isoflurane erroneously selected instead of desflurane. This error can only be detected by a careful checking of the device. PMID- 9732782 TI - [Bilateral sciatica: a predictive sign of extradural hematoma after lumbar disk herniation]. AB - The authors analyse five cases, in order to recognize a sensitive early clinical feature of extradural haematoma following lumbar diskectomy. Neither time delay (time from surgery to the suspicion of the diagnosis) nor pain intensity (a subjective factor which might be obtunded by an analgesic drug) are valuable features. The only reliable predictive sign is bilateral sciatica, which occurred in all cases, before the diagnosis was confirmed by a sensitive and motor deficit in the lower limbs. PMID- 9732783 TI - [First publication of a case of latex allergy]. PMID- 9732784 TI - [Treatment of perioperative acute anemia by human recombinant erythropoietin in a Jehovah's witness]. PMID- 9732785 TI - [Which early sedation for severe brain injured patients?]. PMID- 9732786 TI - [Delayed neurological deficiency after spinal anesthesia in a patient with a history of lymphocytic meningitis]. PMID- 9732787 TI - [Peak effect of ketoprofen is necessary in the treatment of postoperative pain]. PMID- 9732788 TI - [Exertion heat stroke and acute liver insufficiency]. PMID- 9732789 TI - [Diagnosis with DNA probes of ventilator associated acquired pneumonia]. PMID- 9732790 TI - [Medical jargon, in French or English?]. PMID- 9732791 TI - [Risk of confusion in Hypnovel and Anexate packaging (Roche)]. PMID- 9732818 TI - [Four corner bladder base and bladder neck suspension with intraosseous anchorage in the treatment of moderate cystocele]. AB - During the last 10 years the original Pereyra technique of needle bladder neck suspension has been object of more than 36 modifications with the goal to improve long term results and to enhance feasibility. It represents also a part of the so called four corner bladder and bladder neck suspension (anterior suspending sutures) which is at present a reliable and durable manner to manage mild to moderate cystocele; this procedure reestablishes safely and simply support to bladder base, bladder neck and urethra preventing the onset of a denovo stress urinary incontinence. Complications include post-operative pain which could represent a problem in about 16% of the patients: it has been related to the entrapment of the ileoinguinal nerve between prolene sutures and rectus fascia and may be responsible of a delay in the re-establishment of a normal voiding pattern due to the pain elicited during any rectus muscle contraction. We propose a refinement of this procedure which includes the osseous anchoring of the suspending suture through the Mitek G II anchor system. Reduction in postoperative pain and fast recovery of a normal voiding pattern soon after surgery seems to be the most important result of this modification. Osteitis pubis has not been noted. Any improvement in long term durability of the procedure has not yet been determined due to the short follow-up and limited series of cases and the need for subsequent long term follow-up. PMID- 9732819 TI - [Minimally invasive treatment of female urinary incontinence due to sphincter incompetence]. AB - Urinary incontinence is one of the most frequent diseases in female urology. There are hundreds of therapeutic opportunities without sure outcomes, because of lack of the etiologic cause. Transurethral injection has good results in around of the 60% of the cases with minimal invasivity, good patients compliance, ripetitivity of the method. From 1993 to 1997 we treated 44 ladies with urinary incontinence due to low pressure of closure of the sphincter: 13 patients (29.55%) disease free, 17 (38.64%) meliorate. PMID- 9732820 TI - [Improvement of endoscopic resection procedures in relation to the volume of prostatic adenoma]. AB - First choice therapy of obstructive prostatic hyperplasia is still represented by endoscopic transurethral resection of the prostate, despite new techniques for tissue ablation have being proposed. The issue aims to answer to the questions arising about indications of this procedure, and introduces to the knowledge of physical laws which regulate irrigation during and after operation and to the different techniques of irrigating flow derivation whose application reduced morbidity depending on tissue volume and "resection time". Moreover, the new technique of continuous resection with simultaneous suprapubic evacuation of tissue chips (Turbo-TUR) will be illustrated in details, as it permitted to extend indications and advantages of endoscopic surgery to bulky adenomas, electively submitted to open surgery up to now. PMID- 9732821 TI - [Phenotypic changes in mucin-secreting cells in the ileal neobladder mucosa: a metaplastic or precancerous lesion?]. AB - Colonic metaplasia with shifting from sialo-to sulfomucins was observed in 10/18 patients with ileal orthotopic neobladder; their median follow-up was 59 months. There is a significant statistical relationship (p = 0.002) between Colonic Metaplasia and a follow-up longer than 14 months. Diversion Cancer is nowadays a practical problem and probably urologists will be confronted with it in the future more than at present. A modification of established attitudes as regards urinary diversion in standard situations (i.e., 60 over years old patients with invasive bladder cancer) don't seems, at present, justified, although our study confirms the suspicion that ileal neobladder could be considered theoretically at risk for cancer onset. PMID- 9732822 TI - [Prostatic endoprosthesis in the treatment of prostatic obstruction: preliminary experience with the new Trestle model]. AB - The urethral stent is a relatively new treatment modality for benign prostatic hypertrophy ostruction. Although the ultimate prostatic endoprosthesis has yet to be developed, various stents have been produced and investigated with good results. We review the several currently available stents and we report the preliminary clinical experience with a new device (Trestle). PMID- 9732823 TI - Relief of BPO or improvement in quality of life? AB - Benign prostatic hyperplasia (BPH) can cause benign prostatic enlargement with subsequent benign prostatic obstruction (BPO) and lower urinary tract symptoms (LUTS). A reduction in the size of the prostate has long been considered one of the most important treatment goals. However, there is a poor correlation between prostate size and both LUTS and BPO, and between BPO and symptoms. Today, the urologist's primary objectives are to minimize symptoms, relieve BPO and decrease the morbidity associated with BPO. From the patient's point of view, rapid relief of LUTS and immediate improvement in associated quality of life (QOL) are the most important factors. Although there is a good correlation between relief of symptoms (as measured by the International Prostate Symptom Score [I-PSS], for example) and associated improvement in bothersomeness and QOL, particularly that associated with filling ('irritative') symptoms, it is still important to quantify LUTS-related bothersomeness and QOL. Various questionnaires have been developed to measure bothersomeness (e.g. Symptom Problem Index [SPI], Danish PSS [DAN-PSS], International Continence Society BPH Study Group [ICSmale] questionnaire) and QOL (e.g. I-PSS-QOL, BPH Impact Index [BII] and QOL9 BPH specific questionnaire). In addition, the impact of treatment on sexual function should also be taken into account when judging the overall well being or QOL of the patient. A grading system to evaluate the global improvement in patients following treatment has been established. Patients are either graded as showing 'slight', 'moderate' or 'marked' improvement, with the reduction in I-PSS or BII scores required for each classification dependent on baseline symptom severity. Medical treatment strategies designed to alleviate the symptoms of BPH and consequently improve the patient's QOL are now becoming increasingly important. PMID- 9732824 TI - Medical therapy and quality of life. AB - The risk of mortality and long-term morbidity, including loss of sexual function, associated with surgical procedures for symptomatic benign prostatic hyperplasia (BPH) has prompted research into alternative medical therapies. Phytotherapy involves the use of herbal formulations, where the mechanisms of action are usually obscure and although studies have confirmed their effectiveness in symptom relief and improving quality of life (QOL), few placebo-controlled trials exist. Both the 5 alpha-reductase inhibitor finasteride and alpha 1-adrenoceptor antagonists (e.g. alfuzosin, doxazosin, prazosin, tamsulosin and terazosin) have been recommended as appropriate treatment options for patients with lower urinary tract symptoms (LUTS) associated with benign prostatic obstruction (BPO), and their efficacy has been proven in several placebo-controlled trials. Finasteride reduces the static component of BPO--by reducing the size of the prostate--and, as a result, symptom relief is slow (6-12 months) and is predominantly restricted to patients with large prostates (> 40 g). The alpha 1-adrenoceptor antagonists, on the other hand, reduce the dynamic component of obstruction--relaxation of smooth muscle in the prostate, urethra and bladder neck--and provide rapid symptom relief after only a few doses, relieving LUTS more effectively than finasteride and irrespective of prostate size. All of the various alpha 1 adrenoceptor antagonists provide effective and comparable relief of LUTS, and an improvement in bothersomeness and symptom-related QOL. However, it is also important that the therapy is fast acting and acceptable to the patient, in that it does not interfere with other medication or produce unpleasant side effects. These documented properties of the alpha 1A-adrenoceptor antagonists make them an ideal choice for the medical treatment of symptomatic BPH. PMID- 9732826 TI - Symptomatic BPH and hypertension: does comorbidity affect quality of life? AB - Most alpha 1-adrenoceptor antagonists are non-subtype selective and act on smooth muscle in the prostate, as well as in the vascular system and, as such, have effects on blood pressure as well as relieving LUTS (lower urinary tract symptoms) in symptomatic benign prostatic hyperplasia (BPH). As many elderly patients with LUTS also take concomitant antihypertensive therapy, it has been suggested by some that these patients should be treated with an alpha 1 adrenoceptor antagonist that targets both symptomatic BPH and hypertension simultaneously. However, an alternative school of thought believes that hypertension, as a malignant disease, should be treated optimally first, before the LUTS are controlled. Many different classes of antihypertensive drugs have been developed and evidence, with regard to reduction of cardiovascular morbidity and mortality, supports the use of diuretics and beta-blockers in this indication. However, from this point of view, few data support the use of alpha 1 adrenoceptor antagonists in antihypertensive therapy, and studies indicate that elderly patients in particular are prone to orthostatic hypotension and its effects when treated with alpha 1-adrenoceptor antagonists. This, together with the fact that hypertension is such a potentially morbid disease, suggests that alpha 1-adrenoceptor antagonists should not be used as a first line treatment for the treatment of hypertension. Rather, patients with comorbidity should be treated optimally for both diseases, being treated initially for hypertension with the optimal agent available and then with an alpha 1-adrenoceptor antagonist that is not haemodynamically active to target their LUTS. Tamsulosin, a selective alpha 1A-adrenoceptor antagonist, has no clinically significant effect on blood pressure compared with placebo, thus posing less risk for the patient, especially with regard to symptomatic orthostatic hypotension. PMID- 9732825 TI - Cardiovascular effects of alpha-blockers used for the treatment of symptomatic BPH: impact on safety and well-being. AB - Alpha-adrenoceptor antagonists (alpha-blockers) are efficacious in treating lower urinary tract symptoms (LUTS) suggestive of benign prostatic obstruction (BPO), also termed symptomatic benign prostatic hyperplasia (BPH), causing bladder outlet obstruction (BOO). There is little difference among the various alpha blockers in terms of efficacy in treating LUTS. However, conventional quinazoline derivatives such as terazosin, doxazosin and alfuzosin, originally developed for hypertension, have inherent cardiovascular extension effects, which influence the well being and safety of patients with LUTS by impairing physiological blood pressure (BP) control, even when their effect on unchallenged BP may be quite low. Preclinically, tamsulosin, a sulphonamide-substituted phenethylamine, has a relative selectivity for the alpha 1-adrenoceptors of the lower urinary tract. Clinically, this is associated with fewer cardiovascular extension effects with tamsulosin (modified release capsule) 0.4 mg once daily. This allows the use of convenient regimens of 0.4 mg tamsulosin administered once daily after breakfast from initiation of treatment without the need for 'step-up' increases of dose to avoid 'first-dose' hypotension. Extensive investigation, including multiple orthostatic stress testing (which otherwise is unusual in the characterization of alpha-blockers because of their inherent safety), confirms that tamsulosin 0.4 mg definitely carries a lower risk of impaired BP control. PMID- 9732827 TI - Tamsulosin: real life clinical experience in 19,365 patients. AB - OBJECTIVE: To compare the efficacy, global tolerability and blood pressure effects of tamsulosin (0.4 mg once daily) in subgroups of patients with lower urinary tract symptoms (LUTS) suggestive of benign prostatic obstruction (BPO). METHODS: Data from two open-label, observational studies (Study I: 9,507 patients treated for 4 weeks, Study II: 9,858 patients treated for 12 weeks) were analyzed to compare efficacy, global tolerability and effects on blood pressure in subgroups of patients. RESULTS: The efficacy of tamsulosin was largely unaffected by age or previous phytotherapy; in patients with severe symptoms, the efficacy was at least as large as in those with mild or moderate symptoms. More than 90% of patients reported a good or very good tolerability; in a multivariate analysis, this was slightly reduced in patients with concomitant disease but not affected by antihypertensive co-medication or baseline blood pressure. In patients without co-morbidity or co-medication, the tamsulosin-induced blood pressure reductions were similar to those previously reported for placebo treatment; mean additional blood pressure reductions in patients with concomitant disease or medication was not more than 2 mm Hg. Patients who had previously been treated with beta-sitosterol, other plant extracts or finasteride reported tamsulosin to be more effective than their previous treatment. Patients who had previously received beta-sitosterol or other plant extracts rated the global tolerability of tamsulosin to be similar to that of their previous treatment, while those who had previously received finasteride or other alpha 1-adrenoceptor antagonists rated the global tolerability of tamsulosin to be significantly better than that of their previous treatment. CONCLUSIONS: We conclude that tamsulosin is efficacious in all types of patients with LUTS suggestive of BPO. It is globally well tolerated and has marginal effects on blood pressure, including the vast majority of patients with cardiovascular comorbidity, diabetes or on antihypertensive comedication. PMID- 9732829 TI - Cardiovascular effects of alpha-blockers for the treatment of symptomatic BPH. PMID- 9732831 TI - [Detection of soluble interleukin-2 receptor in the serum of patients with non Hodgkin's lymphoma]. AB - BACKGROUND: Patients with non-Hodgkin's lymphoma (NHL) have increased serum levels of soluble interleukin-2 receptor (sCD25). In this study the authors investigate: a) the value of sCD25, compared to other serum markers, as tumor marker, and b) the relationship of the sCD25 with the response to therapy and prognosis. PATIENTS AND METHODS: Serum interleukin-2 receptor (sCD25) levels were measured at diagnosis in 63 patients with NHL (low-grade lymphoma 30 and high grade lymphoma 33). RESULTS: High levels of sCD25 were found in these patients compared to a control group (median 1,757 U/ml vs 385 U/ml; p < 0.0001). Significant differences were also found between the high-grade group and the low grade group, as a whole and within the same Ann Arbor stage. sCD25 showed a correlation coefficient higher than other serum parameters (albumin, LDH, beta 2 microglobulin, uric acid, C-reactive protein) with Ann Arbor stage and with the number of involved lymph nodes or extralymphatic organs. In the high-grade NHL, the median of sCD25 (3,000 U/ml) separates patients with differences in the overall survival (p = 0.0138) and in percentage of complete remisions (p = 0.0079). All the patients with sCD25 < or = 3,000 U/ml reached the remision. The association sCD25 > 3,000 U/ml and albumin < 3.5 g/dl selected to 5 out of 6 patients who failed induction chemotherapy, and only 2 out of 22 who reached the remision. CONCLUSIONS: The sCD25 is the best serum factor for estimating tumor burden in NHL. sCD25 level isolates or associated with albumin provides prognostic information. PMID- 9732832 TI - [Expression of CD44 variant v6 in breast carcinoma and its relationship with other parameters of tumor biology]. AB - BACKGROUND: The variant v6 of CD44 has been associated with metastatic behaviour in neoplasms such as lymphoma, colon carcinoma and breast carcinoma. The expression of CD44v6 in breast carcinoma by flow cytometry and its relationship with other tumor markers is studied in this paper. MATERIAL AND METHODS: The expression of CD44v6 was studied in 46 fresh tissue specimens by flow cytometry using a monoclonal antibody which recognizes the variant v6. Ploidy and cell cycle were also studied by flow-cytometry using propidium iodide labelling. p 53, c-erbB-2 and estrogen receptor expression as well as cell proliferation by Ki-67 staining were performed by immunohistochemistry. RESULTS: Nineteen out of 46 tumors were CD44v6 positive, without correlation with other tumor markers. Levels of CD44v6 in 19 positive samples sligtly correlates with S-phase and hystological grade. CONCLUSIONS: In the present study there was not a correlation among the presence of the variant v6 of CD44 on tumor cells from breast carcinoma and the aggressivity of the tumor. The expression of CD44v6 by flow cytometry does not seem to be a good prognostic marker. PMID- 9732833 TI - [Prognostic factors in uterine sarcomas: a 21-year retrospective study at the Clinic and Provincial Hospital of Barcelona]. AB - BACKGROUND: Uterine sarcomas show low incidence and poor outcome despite the treatment. The prognostic factors for the survival were determined in this study. PATIENTS AND METHODS: Thirty-nine females with sarcoma of the uterus have been studied retrospectively from January 1975 to December 1996. They were treated in the Gynecology and Radiation Oncology Departments at Hospital Clinic i Provincial of Barcelona. Thirty-seven patients had surgery, 22 radiotherapy and 4 chemotherapy. The influence on the disease-specific survival, disease-free survival, local relapse disease-free survival and metastasis disease-free survival from the following pronostic factors was studied: age, pathologic subtype, miometrial invasion, mitosis, vascular and lymphatic invasion, tumor size, stage, radiotherapy and local relapse. RESULTS: 1) The disease-specific survival at 2 and 5 years was 51.5% and 42.5% respectively, and the disease-free survival at 2 and 5 years was 39%; the incidence of local and distant relapses- was 28 and 33%. 2) The multivariate analysis showed that the overall survival and the disease free survival were affected by the vascular invasion (odds ratio [OR] 12 and 32.6, respectively) and the local failure (OR = 3 and 25.5, respectively); the only factor that affected the local relapse-free survival and metastasis free survival was the III and IV stages (OR = 5.6 in both cases). CONCLUSIONS: In uterine sarcomas, the vascular invasion and the local relapse were prognostic factors for overall survival and for disease-free survival. In stages III and IV there was a decrease in the local relapse-free survival and metastasis-free survival. A correlation between vascular invasion and advanced stages was found. The outcome of the uterine sarcomas is poor, local and distant failure being responsible for this bad prognosis. PMID- 9732834 TI - [Gamma-delta peripheral T-cell lymphomas]. PMID- 9732835 TI - [Renopulmonary syndrome]. PMID- 9732836 TI - [Self-medication with antibiotics. The URANO Group]. PMID- 9732838 TI - [Serum sickness caused by minocycline. A rare case]. PMID- 9732837 TI - [The long road of neurotransmission. The third messengers and other transcription factors in the nervous system (and II). Physiopathological and clinical aspects]. PMID- 9732839 TI - [Outbreak of trichinosis caused by T. britovi]. PMID- 9732840 TI - [Arterial hypertension secondary to renal infarction in primary antiphospholipid syndrome]. PMID- 9732841 TI - [Declaration of health for all workers. Accepted during the second meeting of the WHO Collaborating Centers in Occupational Health in October 11-14, 1994 Beijing, China]. PMID- 9732842 TI - [Tissue polypeptide antigen and carcinoembryonic antigen in bladder neoplasms]. AB - The study was designed to evaluate the effect of acetilation phenotype and smoking on the concentration of tissue polypeptide antigen (TPA) and carcinoembryonic antigen (CEA) in serum of patients with neoplasm of bladder. The study covered a group of 76 patients suffering from neoplasm of bladder and a control group composed of 46 persons. Acetylation phenotype as well as TPA and CEA concentrations in serum were determined. In addition, data on smoking habit were collected. The study indicated significant difference in TPA values in both group while there was no difference in regard to CEA values. It was found that in patients with neoplasm of bladder both acetylation phenotype and smoking affect significantly TPA concentration in serum. PMID- 9732844 TI - [Peptic ulcer among workers in the engineering and chemical industries]. AB - Incidence of peptic ulcer among workers employed in the engineering and chemical industries is presented in view of selected environmental factors. The study covered 157 workers with peptic ulcer who reported themselves to outpatient clinics at the Nitrogenous Fertilizers Plant in Pulawy and the Automobile Works in Lublin (5000 and 3252 employees, respectively). The study was conducted over a period of one year. It included: medical examinations, supplementary tests and a questionnaire survey. The questionnaire asked, among others, about working conditions, type of occupation performed and lifestyle. Peptic ulcer was diagnosed in 2.03% of workers engaged in the engineering industry and 1.82% of those working at the chemical plant. The incidence of peptic ulcer was higher among manual workers (2.18%) than in office workers (1.16%). The patients had been exposed to the following environmental factors: excessive noise--21%, vibration--17.2%, commuting--11.5%, extra work--14.5%, conflicts at work--13.5%, conflicts at home--15.3%, irregular meals--21.6%, alcohol intake--22.4%, cigarette smoking--62.4%. The study indicates that peptic ulcer is more often diagnosed in manual than in office workers and its incidence can be associated with working conditions and lifestyle. However, no significant variation in annual morbidity caused by peptic ulcer was observed among workers of the plants under study. PMID- 9732843 TI - [Effect of the cholesterol and protein diet on cytochrome P-450 dependent monooxygenase activity]. AB - The effect of cholesterol and protein diet on mixed-function oxidase activity and desaturation of fatty acids were studid as well as phenobarbital inducibility of those parameters. Investigations were carried out on Wistar adult male rats. The animals were on the cholesterol (0.25%) and protein (32%) diet for 45 days. Cytochrome P-450 and cytochrome b5 contents, NADPH-cytochrome P-450 reductase, NADH-cytochrome b5 reductase, aniline hydroxylase and 4-aminopyrine N-demethylase activities were measured in a hepatic microsomal fraction. The cholesterol diet exerted a negative effect on all parameters except NADH-cytohrom b5 reductase. The protein diet did not change the activity or levels of enzymes under study. The only exception applied to induction of NADH-cytochrome b5 reductase and 4 aminopyrine N-demethylase activities. Simultaneous treatment with phenobarbital had a heterogenous effect depending on the type of diet and parameters examined. PMID- 9732845 TI - [Evaluation of ultraviolet radiation reflectance for human skin]. AB - In a group of 20 males aged 19-20 years ultraviolet radiation reflectance of different wavelength (250-400 nm) from skin surface was measured. It was found that the reflectance increased monotonically with the decrease of wavelength. The reflectance values were compared with the coefficient of relative spectral biological effectiveness and significance of both values for evaluation of human exposure to ultraviolet radiation was discussed. PMID- 9732846 TI - [Disinfectants in health service institutions and frequency of their use]. AB - On the basis of information obtained from sanitary and epidemiological stations places all over the country and from hospitals of the Lodz region it was found that about 60 types of disinfectants are now under use in Poland. The most frequent ones are: lysoformine, chloramine, aldesan, virkon, hypochloride, septil R, cidex, lysol, denatured alcohol, secuspet, aerodesine 2000, desoform, iodoseptane. Some of them contain well known allergenic factors (glutaraldehyde, benzalkonium, hydroquinone, phenol). Bearing in mind an increasing incidence of occupational skin diseases among health service workers it is postulated to continue studies of allergenic properties of disinfectants. PMID- 9732848 TI - [The proposition of a rule for evaluating the percentage of health impairment due to occupationally related myocardial infarction]. AB - The authors propose the principles how to calculate a proportional health impairment due to myocardial infarction. The proposals are based on own experience, literature and regulations. It is suggested that the following should be taken into account for the calculation purposes: case history, resting electrocardiogram, postexercise electrocardiogram, results of Holter's examination, chest x-ray including evaluation of the heart silhoutte and UCG. PMID- 9732847 TI - [Test of electric current perception thresholds at the fingertips--repeatability of the results]. AB - The main aim of present study was to estimate the repeatability of measurements of electric current thresholds performed ten times in the same person. The study covered five persons. Variation in values was connected with individual differences in perception of the stimuli. Coefficients of variation were used as a relative measure of dispersion. Coefficients of variation calculated for individual frequencies were as follows: 7-37% for direct current, 4-19% for alternating current at frequency of 50 Hz, 3-23% for 100 Hz, 5-26% for 400 Hz, 5 23% for 1000 Hz and 4-19% for transient current. Current at frequency of 50 Hz showed the lowest perception threshold and the highest repeatability of values. At frequencies 100, 400 and 1000 Hz values of perception thresholds were higher and more variable. Transient current was sensed at high intensity but it showed small differences in consecutive examinations. PMID- 9732849 TI - [Evaluation of the nervous system in workers as needed for preventive care. Methodical indicators. II. Objective examinations]. AB - A model of medical examinations useful in general evaluation of the nervous system was proposed. It is designed for doctors of occupational health services who perform prophylactic examinations but they are not neurologists. Technically simple elements of routine neurological examinations are selected. They are easy to perform and provide most observations which can be interpreted by a doctor who is not specialised in neurology. PMID- 9732851 TI - [The French system of occupation medicine. II. Evolution of the French system of occupational medicine]. AB - The author presents a discussion on the format of French occupational health system during the eighties. The discussion responded to challenges of civilization changes and new ideas presented by international organization. Exchange of view between supporters and opponents of the existing system produced its new shape based on two fundamental principles: a multidisciplinary character of workers' health care and a greater involvement of doctors in adapting occupational environment to needs of employees. PMID- 9732850 TI - [Nitrosamines]. AB - The paper reviews data on the property and mechanism of toxicologic effects of nitrosamines. Particular attention is turned to metabolism and carcinogenesis of these compounds in view of their concentration in human environment. PMID- 9732852 TI - [Prevention of risk and its control in the work environment. Report from the World Health Organization, 19 October 1994]. AB - On the 19-21 September 1994 an international meeting of experts was convened at the World Health Organization office in Geneva. The result of this meeting was the formation of the PACE working group. PACE stands for "Prevention And Control Exchange". It is programme designed to stimulate the sharing of solutions and control measures in order to reduce occupational hazards. Internationally there is wide agreement on the need for sharing of knowledge and a realisation that a collaborate effort is required. PMID- 9732853 TI - A chronic lack of definition. PMID- 9732854 TI - Arizona professor files lawsuit after being fired for misconduct. PMID- 9732855 TI - Prisoner's DNA database ruled unlawful. PMID- 9732856 TI - NIH institute to work with trial of AIDS vaccine, despite concerns. PMID- 9732857 TI - DuPont opens up access to genetics tool. PMID- 9732858 TI - Outcry as 'scientific' badger cull is launched to target TB. PMID- 9732859 TI - India's short cow drags Roslin Institute into controversy. PMID- 9732860 TI - Tougher crackdown on fraud needed. PMID- 9732861 TI - Modified animal feeds must be put to the test. PMID- 9732862 TI - Evolutionary biology. The secrets of faces. PMID- 9732863 TI - Translation. Cinderella factors have a ball. PMID- 9732864 TI - Immunology. Fetal fascination. PMID- 9732865 TI - Synaptic plasticity. Down with novelty. PMID- 9732866 TI - DNA methylation models histone acetylation. PMID- 9732867 TI - Eukaryotic ribosomes require initiation factors 1 and 1A to locate initiation codons. AB - The scanning model of translation initiation is a coherent description of how eukaryotic ribosomes reach the initiation codon after being recruited to the capped 5' end of messenger RNA. Five eukaryotic initiation factors (eIF 2, 3, 4A, 4B and 4F) with established functions have been assumed to be sufficient to mediate this process. Here we report that eIF1 and eIF1A are also both essential for translation initiation. In their absence, 43S ribosomal preinitiation complexes incubated with ATP, eIF4A, eIF4B and eIF4F bind exclusively to the cap proximal region but are unable to reach the initiation codon. Individually, eIF1A enhances formation of this cap-proximal complex, and eIF1 weakly promotes formation of a 48S ribosomal complex at the initiation codon. These proteins act synergistically to mediate assembly of ribosomal initiation complexes at the initiation codon and dissociate aberrant complexes from the mRNA. PMID- 9732868 TI - A symmetrically pulsed jet of gas from an invisible protostar in Orion. AB - Young stars are thought to accumulate most of their mass through an accretion disk, which channels the gas and dust of a collapsing cloud onto the central protostellar object. The rotational and magnetic forces in the star-disk system often produce high-velocity jets of outflowing gas. These jets can in principle be used to study the accretion and ejection history of the system, which is hidden from direct view by the dust and dense gas of the parent cloud. But the structures of these jets are often too complex to determine which features arise at the source and which are the result of subsequent interactions with the surrounding gas. Here we present infrared observations of a very young jet driven by an invisible protostar in the vicinity of the Horsehead nebula in Orion. These observations reveal a sequence of geyser-like eruptions occurring at quasi regular intervals and with near-perfect mirror symmetry either side of the source. This symmetry is strong evidence that such features must be associated with the formation of the jet, probably related to recurrent or even chaotic instabilities in the accretion disk. PMID- 9732869 TI - Effects of sexual dimorphism on facial attractiveness. AB - Testosterone-dependent secondary sexual characteristics in males may signal immunological competence and are sexually selected for in several species. In humans, oestrogen-dependent characteristics of the female body correlate with health and reproductive fitness and are found attractive. Enhancing the sexual dimorphism of human faces should raise attractiveness by enhancing sex-hormone related cues to youth and fertility in females, and to dominance and immunocompetence in males. Here we report the results of asking subjects to choose the most attractive faces from continua that enhanced or diminished differences between the average shape of female and male faces. As predicted, subjects preferred feminized to average shapes of a female face. This preference applied across UK and Japanese populations but was stronger for within-population judgements, which indicates that attractiveness cues are learned. Subjects preferred feminized to average or masculinized shapes of a male face. Enhancing masculine facial characteristics increased both perceived dominance and negative attributions (for example, coldness or dishonesty) relevant to relationships and paternal investment. These results indicate a selection pressure that limits sexual dimorphism and encourages neoteny in humans. PMID- 9732870 TI - Separate body- and world-referenced representations of visual space in parietal cortex. AB - In order to direct a movement towards a visual stimulus, visual spatial information must be combined with postural information. For example, directing gaze (eye plus head) towards a visible target requires the combination of retinal image location with eye and head position to determine the location of the target relative to the body. Similarly, world-referenced postural information is required to determine where something lies in the world. Posterior parietal neurons recorded in monkeys combine visual information with eye and head position. A population of such cells could make up a distributed representation of target location in an extraretinal frame of reference. However, previous studies have not distinguished between world-referenced and body-referenced signals. Here we report that modulations of visual signals (gain fields) in two adjacent cortical fields, LIP and 7a, are referenced to the body and to the world, respectively. This segregation of spatial information is consistent with a streaming of information, with one path carrying body-referenced information for the control of gaze, and the other carrying world-referenced information for navigation and other tasks that require an absolute frame of reference. PMID- 9732871 TI - Spatial exploration induces a persistent reversal of long-term potentiation in rat hippocampus. AB - Experience-dependent long-lasting increases in excitatory synaptic transmission in the hippocampus are believed to underlie certain types of memory. Whereas stimulation of hippocampal pathways in freely moving rats can readily elicit a long-term potentiation (LTP) of transmission that may last for weeks, previous studies have failed to detect persistent increases in synaptic efficacy after hippocampus-mediated learning. As changes in synaptic efficacy are contingent on the history of plasticity at the synapses, we have examined the effect of experience-dependent hippocampal activation on transmission after the induction of LTP. We show that exploration of a new, non-stressful environment rapidly induces a complete and persistent reversal of the expression of high-frequency stimulation-induced early-phase LTP in the CA1 area of the hippocampus, without affecting baseline transmission in a control pathway. LTP expression is not affected by exploration of familiar environments. We found that spatial exploration affected LTP within a defined time window because neither the induction of LTP nor the maintenance of long-established LTP was blocked. The discovery of a novelty-induced reversal of LTP expression provides strong evidence that extensive long-lasting decreases in synaptic efficacy may act in tandem with enhancements at selected synapses to allow the detection and storage of new information by the hippocampus. PMID- 9732872 TI - Decreased lesion formation in CCR2-/- mice reveals a role for chemokines in the initiation of atherosclerosis. AB - Chemokines are proinflammatory cytokines that function in leukocyte chemoattraction and activation and have recently been shown to block the HIV-1 infection of target cells through interactions with chemokine receptors. In addition to their function in viral disease, chemokines have been implicated in the pathogenesis of atherosclerosis. Expression of the CC chemokine monocyte chemoattractant protein-1 (MCP-1) is upregulated in human atherosclerotic plaques, in arteries of primates on a hypercholesterolaemic diet; and in vascular endothelial and smooth muscle cells exposed to minimally modified lipids. To determine whether MCP-1 is causally related to the development of atherosclerosis, we generated mice that lack CCR2, the receptor for MCP-1 (ref. 7), and crossed them with apolipoprotein (apo) E-null mice which develop severe atherosclerosis. Here we show that the selective absence of CCR2 decreases lesion formation markedly in apoE-/- mice but has no effect on plasma lipid or lipoprotein concentrations. These data reveal a role for MCP-1 in the development of early atherosclerotic lesions and suggest that upregulation of this chemokine by minimally oxidized lipids is an important link between hyperlipidaemia and fatty streak formation. PMID- 9732873 TI - Leptin modulates the T-cell immune response and reverses starvation-induced immunosuppression. AB - Nutritional deprivation suppresses immune function. The cloning of the obese gene and identification of its protein product leptin has provided fundamental insight into the hypothalamic regulation of body weight. Circulating levels of this adipocyte-derived hormone are proportional to fat mass but maybe lowered rapidly by fasting or increased by inflammatory mediators. The impaired T-cell immunity of mice now known to be defective in leptin (ob/ob) or its receptor (db/db), has never been explained. Impaired cell-mediated immunity and reduced levels of leptin are both features of low body weight in humans. Indeed, malnutrition predisposes to death from infectious diseases. We report here that leptin has a specific effect on T-lymphocyte responses, differentially regulating the proliferation of naive and memory T cells. Leptin increased Th1 and suppressed Th2 cytokine production. Administration of leptin to mice reversed the immunosuppressive effects of acute starvation. Our findings suggest a new role for leptin in linking nutritional status to cognate cellular immune function, and provide a molecular mechanism to account for the immune dysfunction observed in starvation. PMID- 9732874 TI - Csk controls antigen receptor-mediated development and selection of T-lineage cells. AB - The development and function of alphabetaT lymphocytes depend on signals derived from pre-T and alphabetaT cell receptors (preTCR and alphabetaTCR) (reviewed in refs 1, 2). The engagement of these receptors leads to the activation of Lck and Fyn, which are protein tyrosine kinases (PTKs) of the Src family. It remains unclear to what extent the activation of Src-family PTKs can direct the differentiation steps triggered by preTCR and alphabetaTCR. Here we show that the inactivation of the negative regulator of Src-family PTKs, carboxy-terminal Src kinase (Csk), in immature thymocytes abrogates the requirement for preTCR, alphabetaTCR and major histocompatibility complex (MHC) class II for the development of CD4+ 8+ double-positive and CD4+ single-positive thymocytes as well as peripheral CD4 alphabetaT-lineage cells. These data show that Csk and its substrates are required to establish preTCR/alphabetaTCR-mediated control over the development of alphabetaT cells. PMID- 9732875 TI - FGF-mediated mesoderm induction involves the Src-family kinase Laloo. AB - During embryogenesis, inductive interactions underlie the development of much of the body plan. In Xenopus laevis, factors secreted from the vegetal pole induce mesoderm in the adjacent marginal zone; members of both the transforming growth factor-beta (TGF-beta) and fibroblast growth factor (FGF) ligand families seem to have critical roles in this process. Here we report the identification and characterization of laloo, a novel participant in the signal transduction cascade linking extracellular, mesoderm-inducing signals to the nucleus, where alteration of cell fate is driven by changes in gene expression. Overexpression of laloo, a member of the Src-related gene family, in Xenopus embryos gives rise to ectopic posterior structures that frequently contain axial tissue. Laloo induces mesoderm in Xenopus ectodermal explants; this induction is blocked by reagents that disrupt the FGF signalling pathway. Conversely, expression of a dominant inhibitory Laloo mutant blocks mesoderm induction by FGF and causes severe posterior truncations in vivo. This work provides the first evidence that a Src related kinase is involved in vertebrate mesoderm induction. PMID- 9732876 TI - Smad3 and Smad4 cooperate with c-Jun/c-Fos to mediate TGF-beta-induced transcription. AB - Smad proteins transduce signals for transforming growth factor-beta (TGF-beta) related factors. Smad proteins activated by receptors for TGF-beta form complexes with Smad4. These complexes are translocated into the nucleus and regulate ligand induced gene transcription. 12-O-tetradecanoyl-13-acetate (TPA)-responsive gene promoter elements (TREs) are involved in the transcriptional responses of several genes to TGF-beta (refs 5-8). AP-1 transcription factors, composed of c-Jun and c Fos, bind to and direct transcription from TREs, which are therefore known as AP1 binding sites. Here we show that Smad3 interacts directly with the TRE and that Smad3 and Smad4 can activate TGF-beta-inducible transcription from the TRE in the absence of c-Jun and c-Fos. Smad3 and Smad4 also act together with c-Jun and c Fos to activate transcription in response to TGF-beta, through a TGF-beta inducible association of c-Jun with Smad3 and an interaction of Smad3 and c-Fos. These interactions complement interactions between c-Jun and c-Fos, and between Smad3 and Smad4. This mechanism of transcriptional activation by TGF-beta, through functional and physical interactions between Smad3-Smad4 and c-Jun-c-Fos, shows that Smad signalling and MAPK/JNK signalling converge at AP1-binding promoter sites. PMID- 9732877 TI - Importance of diabetes mellitus and systemic hypertension rather than completeness of revascularization in determining long-term outcome after coronary balloon angioplasty (the LDCMC registry). Lady Davis Carmel Medical Center. AB - The study examined the 10-year outcome in a cohort of 227 unselected, consecutive patients (age 58+/-10 years) undergoing coronary balloon angioplasty between 1984 and 1986 and followed in a single cardiac center (Lady Davis Carmel Medical Center registry). In particular, we sought to identify the relative importance of the systemic risk factors diabetes and hypertension and the extent of coronary disease as opposed to procedure-related technical variables, the immediate success of the procedure, or completeness of revascularization. By life-table analysis (99% follow-up), 94% of the patients were alive at 5 years, and 77% at 10 years after angioplasty. Ten-year survival was reduced in patients with diabetes mellitus (59% vs 83%, p = 0.0008), in patients with previous myocardial infarction (68% vs 85%, p = 0.01), in patients with ejection fraction <50% (55% vs 82%, p = 0.005), and in patients with 3-vessel disease (58% vs 84% and 86% for 1- and 2-vessel disease, respectively, p = 0.04). Diabetes mellitus was the major independent predictor of poor survival (adjusted odds ratio 3.1, 95% confidence interval 1.55 to 6.19, p = 0.001). Survival at 10 years was identical in 199 patients in whom angioplasty was complete and in 25 in whom the balloon catheter did not cross the lesion, although bypass surgery was more frequent in the latter group (45% vs 21%, p = 0.001). Incomplete revascularization did not predict poor survival (72% vs 79% with complete angioplasty, p = NS). Event-free survival at 10 years for the whole group was 29%, and 49% of patients survived with no event other than a single repeat angioplasty procedure. Multivessel disease, hypertension, and diabetes mellitus were independent predictors of decreased event-free survival, but incomplete revascularization was not. Thus, long-term outcome after coronary balloon angioplasty was related to diabetes mellitus, systemic hypertension, and extent of coronary disease, but not to the immediate success of the procedure or completeness of revascularization. PMID- 9732878 TI - Incidence, consequences, and risk factors of early reocclusion after primary and/or rescue percutaneous transluminal coronary angioplasty for acute myocardial infarction. AB - Percutaneous transluminal coronary angioplasty (PTCA) for acute myocardial infarction (AMI) achieves high patency rates. Conversely, it has been shown that after thrombolysis, early reocclusion of the infarct-related artery (IRA) is associated with substantial morbidity and mortality. The aim of this retrospective study was to study the incidence, prognostic implications, and clinical risk factors for in-hospital reocclusion of the IRA after successful emergency PTCA for AMI. We studied 399 consecutive patients (aged 59+/-14 years, 52% with anterior wall infarction) admitted <6 hours after AMI onset, of whom 374 (94%) were successfully treated with primary (n = 297) or rescue (n = 77) PTCA, with a stenting rate of 8%. Predischarge angiography was performed in 306 (82%). Early reocclusion of the IRA occurred in 28 patients (9%) and was silent in 6 (2%). The reocclusion rate was 10% for primary PTCA and 8% for rescue PTCA (p = NS). Twenty-two of 28 patients (6%) underwent repeat emergency coronary angiography because of early recurrent ischemia and most (n = 18) were treated with emergency PTCA. Early recurrent ischemia occurred mostly (86%) within 5 days of AMI onset. There was a higher prevalence of on-site hemorrhage (18% vs 5%, p = 0.007), blood transfusion (11% vs 2%, p = 0.01), pulmonary edema (21% vs 4%, p <0.01), and in-hospital death (21% vs 1%, p = 0.0001) in patients with predischarge reocclusion. On multivariate analysis, cardiogenic shock on admission and absence of dyslipidemia were strong and independent predictors (p = 0.01) of IRA reocclusion. In conclusion, early reocclusion after emergency PTCA occurred in 9% of the patients and was associated with substantial morbidity and mortality. This warrants attempts to decrease its incidence, e.g., with more frequent use of stents. PMID- 9732880 TI - Usefulness of isometric hand grip exercise in detecting coronary artery disease during dobutamine atropine stress echocardiography in patients with either stable angina pectoris or another type of positive stress test. AB - Dobutamine atropine stress echocardiography (DASE) detects coronary artery disease (CAD) by increasing myocardial oxygen demand causing ischemia. The sensitivity of the test for detection of CAD is reduced in patients with submaximal stress. We hypothesized that increasing cardiac work load by adding isometric exercise would improve the detection of ischemia during DASE. We studied 31 patients, mean age 57+/-11 years, with angiographically documented CAD. Patients underwent DASE using incremental dobutamine doses from 5 to 40 microg/kg/min, followed by atropine if peak heart rate was <85% of predicted maximal. Hand grip was then performed for 2 minutes at 33% of maximal voluntary contraction, while dobutamine infusion was maintained at the peak dose. The addition of hand grip during dobutamine stress was associated with a significant increase in systolic blood pressure (143+/-21 vs 164+/-24 mm Hg, p = 0.001) and left ventricular end-systolic circumferential wall stress (72+/-30 x 10(3) dynes/cm2 vs 132+/-34 x 10(3) dynes/cm2, p = 0.004). Wall motion score index increased from 1.0 at rest to 1.15+/-0.18 with dobutamine (p = 0.0004 vs rest), and increased further to 1.29+/-0.22 with the addition of hand grip (p = 0.004 vs dobutamine). Ischemia was detected in 19 patients (62%) with dobutamine-atropine stress alone and in 25 (83%) after the addition of hand grip (p <0.05). The addition of hand grip during DASE is feasible, and improves the detection of myocardial ischemia. PMID- 9732879 TI - Metabolic changes in hibernating myocardium after percutaneous transluminal coronary angioplasty and the relation between recovery in left ventricular function and free fatty acid metabolism. AB - To elucidate the changes in oxidative metabolism in hibernating myocardium after coronary revascularization, we performed myocardial single-photon emission computed tomography with a free fatty acid analog, I-123 beta methyliodophenylpentadecanoic acid (BMIPP), and thallium-201 before and 1 month after percutaneous transluminal coronary angioplasty (PTCA) in 11 patients with angina pectoris caused by single artery stenosis. All patients had improvement in wall motion after PTCA at the region with coronary stenosis; the wall motion abnormality score evaluated by left ventriculography decreased from 5.5+/-0.8 (mean +/- SE) to 2.1+/-0.9, p <0.01) after PTCA. The defect score of I-123 BMIPP images was significantly larger than that of thallium-201 images either before (14+/-1.3 vs 8.9+/-1.1, p <0.01) or 1 month after (7.4+/-1.5 vs 3.7+/-0.8, p <0.01) PTCA. The decrease in the defect score of both images was significant (p <0.01). Changes in the wall motion abnormality score showed a significant correlation with both the change in the defect score of thallium-201 images (r = 0.58, p < 0.01) and that of I-123 BMIPP images (r = 0.75, p <0.01). These results indicate that the metabolism of free fatty acid is impaired in hibernating myocardium, and that improvement in left ventricular function after successful PTCA is strongly associated with the recovery of oxidative metabolism. PMID- 9732881 TI - Usefulness of exercise echocardiography in predicting cardiac events in an outpatient population. AB - The prognostic value of exercise echocardiography in an outpatient population is not well defined. A total of 1,020 consecutive patients referred for exercise echocardiography in an ambulatory care setting were studied by reviewing their medical records and exercise echocardiographic data. Of these, 71 (7%) were excluded due to technically inadequate tests, leaving 949 patients who were included in the analysis. A positive exercise echocardiogram (EE) was defined as an appearance of a new wall motion abnormality or worsening of a baseline abnormality. Cardiac events, defined as myocardial infarction, coronary angioplasty, coronary bypass surgery, and death, were documented during a 12 month follow-up period. Cardiac events occurred in 17% of patients (26 of 152) with a positive exercise echocardiogram (EE) and in 2.5% (20 of 797) with a negative EE (p <0.001). The incidence of myocardial infarction (2.6% vs 0.4%, p <0.02), coronary angioplasty (7% vs 1%, p <0.001), and coronary bypass surgery (9% vs 1%, p <0.001) were higher in patients with a positive versus a negative EE. There was 1 death in the positive study group and none in the negative group. Significant independent variables (p <0.05) that predicted cardiac events included a positive exercise electrocardiogram, history of coronary angioplasty, nonspecific ST-T changes on the baseline electrocardiogram, double product <25,000, men, chest pain on exercise test, and a positive exercise electrocardiogram. On a stepwise logistic regression model, exercise echocardiography emerged as an independent predictor of future cardiac events in an outpatient population. This predictive value was enhanced in the presence of a positive exercise electrocardiogram compared with a negative exercise electrocardiogram (24.2% vs 7.9%, p <0.03). Our study suggests that exercise echocardiography is an independent predictor of future cardiac events in an outpatient population. PMID- 9732882 TI - Developing and testing a system to improve the quality of heparin anticoagulation in patients with acute cardiac syndromes. AB - We have taken a stepwise approach to improving the dosing of continuous intravenous heparin in patients with acute coronary syndromes. Our primary objective was to use computer modeling to develop a nomogram for managing heparin therapy and to put in place a continuous quality monitoring system to evaluate the nomogram's effectiveness. We prospectively collected data on 41 patients with unstable angina or myocardial infarction who were treated with heparin. Their response to heparin was computer modeled and the dose to achieve an activated partial thromboplastin time (aPTT) ratio of 2.0 was established. This dose was regressed against all demographic characteristics to establish predictors of heparin dose (phase I). The regression formula was used prospectively in 110 patients to initiate the infusion rate of heparin and a bolus dose to achieve an aPTT ratio of 2.5. Subsequent dosage adjustments were achieved by computer modeling the patient's aPTT response (phase II). A nomogram was developed that simulated the decisions achieved using computer-assisted methods. This was retrospectively tested and then prospectively tested in 50 patients using nursing staff (phase IV). The nomogram was then made generally available (phase IV) and has been tested in an additional 310 patients. Phase I: Of the original 41 patients, 32% of the aPTT ratios were in the therapeutic range, 36% were supratherapeutic, and 32% were subtherapeutic after the first 24 hours. Phases II and III resulted in 85% of the aPTT ratios between 1.5 and 2.5 at 24 hours. Phase 4 had similar results in 310 patients. The use of computer-assisted or a computer generated nomogram to adjust heparin therapy results in better control of heparin therapy than using standard methods. PMID- 9732883 TI - Risk of thromboembolic events in patients with atrial flutter. AB - Based on multiple studies, clear, guided anticoagulation therapy is recommended for patients with atrial fibrillation. The value of anticoagulation therapy in patients with atrial flutter, however, is less well established. Little is known about the incidence of thromboembolism in patients with atrial flutter. We evaluated the risk of thromboembolism in 191 consecutive unselected patients referred for treatment of atrial flutter. A history of embolic events was noted in 11 patients. Acute embolism (<48 hours) occurred in 4 patients (3 after direct current cardioversion, 1 after catheter ablation). During follow-up of 26+/-18 months, 9 patients experienced thromboembolic events. During the follow-up, the overall embolic event rate (including acute embolism and thromboembolic events during follow-up) was 7 % in this patient population. Risk indicators for an embolic event in an univariate analysis were organic heart disease (p = 0.037), depressed left ventricular function (p = 0.02), history of systemic hypertension (p = 0.004), and diabetes mellitus (p = 0.0038). Using multivariate analysis, a history of hypertension was the only independent predictor for elevated embolic risk in this patient population (odds ratio = 6.5; 95% confidence intervals 1.5 to 45). Thus, the thromboembolic risk is higher than previously recognized for patients with atrial flutter. Anticoagulation therapy may decrease this risk. PMID- 9732884 TI - Comparison between propafenone and digoxin administered intravenously to patients with acute atrial fibrillation. PAFIT-3 Investigators. The Propafenone in Atrial Fibrillation Italian Trial. AB - In recent-onset atrial fibrillation, intravenous propafenone has been shown to effectively restore sinus rhythm, whereas the efficacy of intravenous digoxin has been questioned. We directly compared these 2 drugs and placebo in acute atrial fibrillation. One hundred twenty-three patients with atrial fibrillation lasting <72 hours were randomized to a 10-minute intravenous infusion of either propafenone (2 mg/kg, 41 patients) or digoxin (0.007 mg/kg, 40 patients) or placebo (42 patients). After 1 hour, nonconverted propafenone or digoxin patients were switched to the alternative drug, while nonconverted placebo patients were randomized to either propafenone or digoxin. The observation time ended 1 hour later. By 1 hour, conversion rates were 49% in the propafenone group, 32% in the digoxin group (p = 0.12), and 14% in placebo group (p <0.001 vs propafenone, p = 0.08 vs digoxin). After crossover, digoxin converted 5% of propafenone patients, while propafenone converted 48% of digoxin patients (p <0.05). In the 36 nonconverted placebo patients, sinus rhythm was obtained in 53% of cases with propafenone, and in 5% with digoxin (p < 0.05). Globally, among the 116 patients who received a drug as first treatment, 30 of 60 patients (50%) were converted by propafenone versus 14 of 56 (25%) by digoxin (p <0.01) (odds ratio 2.0, 95% confidence interval 1.19 to 3.36). In nonconverters, the ventricular rate reduction was faster (15 vs 45 minutes) and more prominent (-24% vs -14%) with propafenone than with digoxin. In conclusion, intravenous propafenone terminates atrial fibrillation more effectively than either placebo or intravenous digoxin. In addition, in nonconverted patients, it obtains a more rapid and marked control of the ventricular rate. PMID- 9732885 TI - Radiofrequency catheter ablation versus medical therapy for initial treatment of supraventricular tachycardia and its impact on quality of life and healthcare costs. AB - We prospectively compared the impact on quality of life and cost effectiveness between ablation and medication as an initial strategy for patients with paroxysmal supraventricular tachycardia (SVT). Seventy-nine consecutive patients with newly documented paroxysmal SVT were treated with either ablation or medication. Health surveys (SF-36 and disease-specific questions) were obtained at baseline and after 12 months of follow up. Cost of health care utilization for the 6 months before and after treatment were measured. Both medication and ablation improved quality of life. However, ablation improved quality of life in more general health categories than medication. At follow up, ablation was associated with significantly improved quality of life in the bodily pain (63+/ 24 vs 81+/-20, p <0.005), general health (69+/-21 vs 79+/-21, p <0.05), vitality (55+/-21 vs 66+/-22, p <0.05), and role emotion (78+/-36 vs 94+/-17, p <0.05) categories when compared with medication. Although both medication and ablation decreased frequency of disease-specific symptoms, ablation resulted in complete amelioration of symptoms in more patients (33% vs 74%). Potential long-term costs were similar for medication and ablation. In conclusion, ablation improves health related quality of life to a greater extent, and in more aspects of general and disease-specific health than medication. PMID- 9732886 TI - Electrocardiographic conduction disturbances in association with low-level lead exposure (the Normative Aging Study). AB - Recent research indicates that cumulative exposure to lead may be more toxic than previously thought. This study was undertaken to examine the relation of low level lead exposure to electrocardiographic (ECG) conduction disturbances among 775 men who participated in the Normative Aging Study (average age 68 years; range 48 to 93). We used K-x-ray fluorescence to measure lead levels in the tibia and patella, and graphite furnace atomic absorption spectroscopy to measure blood lead levels. The mean (SD) values for blood lead, tibia lead, and patella lead were 5.8 (3.4) microg/dl, 22.2 (13.4) microg/g, and 30.8 (19.2) microg/g, respectively. Bone lead levels were found to be positively associated with heart rate-corrected QT and QRS intervals, especially in younger men. Specifically, in men <65 years of age, a 10 microg/g increase in tibia lead was associated with an increase in the QT interval of 5.03 ms (95% confidence interval [CI], 0.83 to 9.22) and with an increase in the QRS interval of 4.83 ms (95% CI, 1.83 to 7.83) in multivariate regression models. In addition, an elevated bone lead level was found to be positively associated with an increased risk of intraventricular block in men <65 years of age and with an increased risk of atrioventricular (AV) block in men > or = 65 years of age. After adjustment for age and for serum high density lipoprotein (HDL) level, a 10 microg/g increase in tibia lead was associated with an odds ratio (OR) of 2.23 (95% CI, 1.28 to 3.90) for intraventricular block in men <65 years of age and with an OR of 1.22 (95% CI, 1.02 to 1.47) for AV block in men > or = 65 years of age. Blood lead level was not associated with any of the ECG outcomes examined. The results suggest that cumulative exposure to lead, even at low levels, may depress cardiac conduction. PMID- 9732887 TI - Changes in pacing lead impedance over time predict lead failure. AB - It has been suggested that a decrease in lead impedance may predict pacing lead failure, but there is limited prospective data about the relation of changes in lead impedance over time to lead performance. We monitored changes in lead impedance through implantable pulse generators with real-time telemetry data capability in 105 patients with Medtronic 4012 leads (n = 38) and Medtronic 4004 leads (n = 67). Pacing lead failure was documented by serial ambulatory electrocardiographic monitoring or intensified pacemaker clinic surveillance. A significant decrease in lead impedance was observed in patients with Medtronic 4012 and Medtronic 4004 leads with documented lead failure, whereas lead impedance remained stable over time in patients without documented lead failure. The sensitivity and specificity of a lead impedance decrease of > or =15% to predict lead failure were 69% and 70%, respectively. The sensitivity and specificity of a lead impedance decrease of > or =30% to predict lead failure were 36% and 90%, respectively. The positive and negative predictive values for a lead impedance decrease of > or =15% were 54% and 81%, respectively, and for a lead impedance decrease of > or = 30% were 65% and 73%, respectively. Thus, small decreases in lead impedance may identify failing leads. Serial measurement of pacing lead impedance over time is a useful tool to monitor pacing lead performance. PMID- 9732888 TI - Comparison in systemic hypertension of left ventricular mass and geometry with systolic and diastolic function in patients <65 to > or = 65 years of age. AB - Previous studies have differed on the independent effect of age and gender to left ventricular (LV) mass. Data on ventricular remodeling in hypertensive patients > or = 65 years of age is lacking. Similarly, the systolic and diastolic interaction in older hypertensives is not well defined. In a prospective study, we examined the relation of LV mass, relative wall thickness, and systolic and diastolic interaction in 508 hypertensive patients between 50 and 80 years of age who were divided according to age (<65 and > or = 65 years) and gender. LV mass, geometric classification, systolic wall stress, and Doppler filling were obtained according to standard Doppler echocardiographic criteria. In men, most measurements were similarly distributed. However, women > or = 65 years of age had smaller LV systolic dimensions, thicker ventricular septums, higher endocardial and midwall fractional shortenings, and lower end-systolic wall stress. Although LV mass was higher in men, there was no age difference within the same sex. The most common LV geometric remodeling was increased relative wall thickness in the form of concentric hypertrophy or concentric remodeled. The predominant mitral flow pattern was "impaired relaxation"; however, older patients had even shorter E waves, taller A waves, and lower E/A ratios. Thus, patients > or = 65 years of age had an even higher prevalence of this pattern (men, 89% vs 73%, p <0.001, and women, 91% vs 77%, p <0.001). Delayed LV relaxation with preservation of systolic ejection indexes is an early abnormality in essential hypertension, which lasts an undetermined time with further progression as patients aged. As a result, hypertensive patients > or = 65 years of age had the most pronounced structural and functional changes, an observation particularly noted in women. In those > or = 65 years, data from the Doppler E wave and A wave do not distinguish the physiologic process of aging from the pathologic changes of pressure overload. PMID- 9732889 TI - The influence of basal nitric oxide activity on pulmonary vascular resistance in patients with congestive heart failure. AB - Increased pulmonary resistance may reduce survival and treatment options in patients with congestive heart failure. Nitric oxide (NO) is a determinant of normal pulmonary resistance vessel tone. We tested the hypothesis that loss of NO function contributes to increased pulmonary vascular resistance index (PVRI) in congestive heart failure. Pulmonary arterial resistance vessel function was studied in 25 conscious adults. Three groups were studied: 8 controls, 9 patients with congestive heart failure and normal PVRI, and 8 patients with congestive heart failure and raised PVRI. Segmental arterial flow was determined with a Doppler wire and quantitative angiography. NG-monomethyl-L-arginine (L-NMMA) was used to inhibit NO, whereas phenylephrine was used as an endothelium-independent control. The response to inhibition of NO with L-NMMA was less in patients with congestive heart failure and elevated PVRI than in patients with congestive heart failure and normal PVRI (p <0.05). The difference in response between the congestive heart failure groups was specific to NO-dependent regulation because the response to the endothelium-independent constrictor phenylephrine was not different (p = 0.92). There was no difference in response to L-NMMA between controls and patients with congestive heart failure and normal PVRI. The response to L-NMMA correlated to PVRI. In adults with congestive heart failure, NO appears to play an important role in maintaining normal pulmonary resistance. PMID- 9732890 TI - Increased ventricular repolarization inhomogeneity during postural changes in patients with syndrome X. AB - The interlead variation in QT interval (QT dispersion) can be used to assess regional inhomogeneity of ventricular repolarization under a variety of conditions, including stress. Patients with syndrome X may have increased sympathetic activity that could change QT interval regionally and give rise to an increase in QT dispersion under exercise testing. To test the hypothesis, 26 consecutive patients with syndrome X (group 1) were studied. Two additional groups matched in terms of age, sex, and left ventricular mass index consisting of 26 nonconsecutive patients with coronary artery disease (group 2) and 20 normal subjects (group 3) were studied for comparison. Standing induced a significantly higher increase of heart rate in group 1 than in groups 2 and 3 (7.5+/-6.0 vs 4.0+/-6.3 and 1.1+/-3.6 beats/min; p = 0.05 and 0.003, respectively). There were significant differences in QT dispersion between groups 1 and 2 on upright standing (48+/-12 vs 34+/-14 ms, p = 0.0003), but not at baseline (33+/-14 vs 38+/-11 ms, p = NS) or at peak exercise (38+/-9 vs 38+/-9 ms, p = NS). Results did not change when QTc dispersion was substituted for QT dispersion. From a conditional multivariate logistic regression analysis, the only independent predictor of occurrence of syndrome X on upright standing was QTc dispersion (odds ratio = 1.255, p = 0.01). Electrocardiographic QTc dispersion provides important clinical information. Patients with syndrome X had a higher increase of heart rate and QTc dispersion in response to standing from the supine position compared with patients with coronary artery disease and normal subjects. PMID- 9732891 TI - Relation of age to left ventricular function in clinically normal adults. AB - The extent to which age, independent of cardiovascular diseases, influences left ventricular (LV) function in adults is uncertain. Echocardiograms and simultaneous arterial pressure in 464 clinically normal adults aged 16 to 88 years were used to measure LV dimensions, endocardial and midwall LV fractional shortening, stroke volume, cardiac output, and circumferential end-systolic stress. The ratios of observed endocardial and midwall shortening to values predicted for observed end-systolic stress were used as measures of chamber and myocardial function. LV endocardial shortening increased slightly with age, as did an index of LV chamber performance, the end-systolic stress/volume index ratio (r = 0.11, p = 0.019, and r = 0.20, p <0.001). However, when age-related increases in LV wall thickness and blood pressure were controlled for by examining afterload-corrected endocardial shortening, no age relation was detected. Weak age-related declines were observed in midwall shortening (r = 0.09, p = 0.043) and afterload-corrected midwall shortening (r = -0.12, p <0.01). Cardiac index decreased slightly with advancing age (r = -0.14, mean -6.7 ml/min/m2/ year, p = 0.003). Total peripheral resistance and the pulse pressure/stroke volume ratio, a measure of arterial stiffness, increased more strongly with age (r = 0.27 and 0.38, both p <0.001). Thus, LV pump performance at rest measured by cardiac index is slightly lower in older than in younger clinically normal adults. Endocardial fractional shortening was slightly higher in older subjects, but the physiologically more appropriate midwall measures of myocardial function decreased slightly. The observed change in LV pump performance was related to smaller LV chamber size and higher total peripheral resistance in older subjects. PMID- 9732892 TI - Comparison of cardiac findings at necropsy in octogenarians, nonagenarians, and centenarians. AB - Certain clinical and necropsy cardiac findings are described and compared in 391 octogenarians (80%), 93 nonagenarians (19%), and in 6 centenarians (1%). The number of men and women was similar (248 [51%] and 242 [49%]). The cause of death was cardiac in 228 patients (47%), vascular but noncardiac in 71 (14%), and noncardiac and nonvascular in 191 (39%). The frequency of a cardiac condition causing death decreased with increasing age groups (51% vs 32% vs 0), and the frequency of a noncardiac, nonvascular condition causing death increased with increasing age groups (36% vs 47% vs 100%). Among the cardiac conditions causing death, coronary artery disease was found in 62% of cases (141 of 228), aortic valve stenosis in 16% (36 of 228), and cardiac amyloidosis in 10% of cases (22 of 228). Calcific deposits were found at necropsy in the coronary arteries in 81% of the patients (398 of 490), in the aortic valve in 47% (228 of 490), in the mitral annular area in 39% of the patients (190 of 490), and in 1 or both left ventricular papillary muscles in 25% of the patients (122 of 490). The calcific deposits tended to be less frequent in the octogenarians. Three hundred (61%) of the 490 patients had > or = 1 major coronary arteries narrowed > 75% in cross sectional area by plaque and the percent of patients in each of the 3 age groups and the percent of coronary arteries significantly narrowed in each of the 3 age groups were similar. PMID- 9732893 TI - Influence of guidewire and catheter type on the frequency of cerebral microembolic signals during left heart catheterization. AB - Cerebral embolization is a serious complication during diagnostic heart catheterization. To date there have been no studies to determine whether the technique and the catheter type influence the frequency of cerebral microembolic signals (MES's) during left ventricular catheterization. Twenty-two patients had a leading straight tip guidewire protruding 5 to 10 cm outside the coronary catheters when the latter was advanced over the aortic arch (group A), whereas in 21 patients the guidewire was withdrawn in the descending part of the aorta (group B). Transcranial Doppler of the left middle cerebral artery was performed to monitor the number of cerebral MES's. When a protruding guidewire was used to advance the coronary catheters over the aortic arch, MES's were detected in 86% of the patients compared with 29% when the catheters were advanced without a guidewire (relative risk = 4.6, p = 0.00001). The number of MES's per patient also was significantly higher when a guidewire was used (median 9 vs 0) (p = 0.000004). In group A, a higher number of MES's was detected when a right Judkins catheter was advanced over the aortic arch than when a left Judkins catheter was advanced (median 6.5 vs 1) (p = 0.0005) and in patients who previously had a myocardial infarction than in those who had not (median 1 1 vs 4) (p = 0.007). This study strongly suggests that the risk of embolization is greater when straight tip guidewires are used to advance catheters over the aortic arch during left ventricular heart catheterization, especially in patients with a history of myocardial infarction. PMID- 9732894 TI - Thrombolism with atrial flutter. AB - These data provide convincing evidence that patients with atrial flutter have a significant thromboembolic risk. Their anticoagulation should be managed in the same manner as patients with atrial fibrillation. PMID- 9732895 TI - Robert McKinnon Califf, MD: a conversation with the editor. Interview by William Clifford Roberts. PMID- 9732896 TI - Initial results of laser-based percutaneous myocardial revascularization for angina pectoris. AB - Results of a 30-patient pilot study of a recently developed percutaneous myocardial revascularization approach are described. The feasibility and positive safety profile of percutaneous myocardial revascularization are clearly demonstrated, with no mortality associated with the treatment or in the immediate post-treatment period and an incidence of only 1 major complication. PMID- 9732897 TI - Prognosis of medically stabilized unstable angina pectoris with a negative exercise test. AB - Three hundred twenty seven patients with medically stabilized unstable angina and a negative exercise test were followed-up during a mean of 39 months. Male gender, diabetes mellitus, and previous myocardial infarction, but not exercise parameters, were predictors of death or acute myocardial infarction. PMID- 9732898 TI - Diagnosing coronary arterial stent thrombosis and arterial closure. AB - This single-center review of a consecutive series of patients requiring reexamination by angiography within 1 week of a coronary stent placement due to chest pain reveals that patients treated with a poststent anticoagulation regimen of warfarin and aspirin, and those with lower poststent deployment dilation pressures, have an increased risk of subacute stent thrombosis. Repeat cardiac catheterization within the first week after coronary artery stent implantation should be reserved for patients with significant electrocardiographic changes. PMID- 9732900 TI - Intravascular ultrasound findings in stenting of unprotected left main coronary artery stenosis. AB - We evaluated the role of intravascular ultrasound (IVUS) in 16 patients with unprotected left main coronary artery (LMCA) stenting compared with 80 patients with other (non-LMCA) native coronary artery stenting and found that (1) additional high-pressure or larger size balloon dilations were more frequently performed in LMCA stenting than in non-LMCA stenting (p <0.05) and (2) after IVUS guided stent implantation, minimum lumen area was > or = 9 mm2 in 88% of patients who underwent LMCA stenting and in 19% of those who underwent non-LMCA stenting (p <0.001). IVUS guidance may be a more important adjunctive imaging modality in the stenting of unprotected LMCA stenoses than in stenting of non-LMCA stenoses. PMID- 9732899 TI - Underutilization of lipid-lowering drugs in older persons with prior myocardial infarction and a serum low-density lipoprotein cholesterol > 125 mg/dl. AB - A prospective study of 500 consecutive persons (aged 60 years with Q-wave myocardial infarction admitted to a long-term health care facility) investigated the prevalence of the use of a lipid-lowering drug at the time of admission in persons with a fasting serum low-density lipoprotein (LDL) cholesterol >125 mg/dl measured the day after admission. The prevalence of a lipid-lowering drug in persons with myocardial infarction and an LDL cholesterol >125 mg/dl was 7% (11 of 153 persons) in persons 60 to 80 years of age and 3% (6 of 182 persons) in persons 81 to 100 years of age. PMID- 9732901 TI - Local administration of L-703,081 using a composite polymeric stent reduces platelet deposition in canine coronary arteries. AB - We compared the effect on platelet deposition of the glycoprotein IIb/IIIa receptor antagonist L-703,081, administered locally via a drug delivery stent, with that of a standard metal stent in a canine coronary model. There was a significant reduction in platelet deposition using the L-703,081-impregnated stent compared with the bare metal stent. This study demonstrates an alternative route of delivery of GPIIb/IIIa antagonists with potential advantages over systemic administration. PMID- 9732902 TI - Clinical reliability of single-lead VDD pacing from evaluation of P-wave sensing under dynamic conditions. AB - In a group of 20 patients implanted with a single-lead VDD pacing system, a wide interindividual variability was found in P-wave amplitude changes occurring under dynamic conditions, even though they were of minor clinical relevance because constant atrial tracking was maintained. PMID- 9732903 TI - Effects of adenosine on retrograde refractoriness of accessory atrioventricular connections. AB - Ventricular premature stimuli were used to demonstrate adenosine-mediated decreases in the retrograde refractoriness of accessory atrioventricular connections. This response is consistent with the concept that accessory atrioventricular connections have electrophysiologic properties that are similar to those of atrial myocardium. PMID- 9732904 TI - Estimating the proportion of post-myocardial infarction patients who may benefit from prophylactic implantable defibrillator placement from analysis of the CAST registry. Cardiac Arrhythmia Suppression Trial. AB - We defined the proportion of post-myocardial infarction patients who would have been eligible for the Multicenter Automatic Defibrillator Implantation Trial (MADIT) from a population of 94,797 patients with myocardial infarction entered into the Cardiac Arrhythmia Suppression Trial Registry. From this large population, only between 0.3% to 1.7% would have met strict eligibility criteria for MADIT. PMID- 9732905 TI - Impact of a nurse practitioner on the cost of managing inpatients with heart failure. AB - We examined the impact on hospital costs of having a nurse practitioner manage uncomplicated patients hospitalized for decompensated heart failure. This strategy was associated with a significant decrease in total hospital costs ($6,659+/-5,843 vs $5,211+/-4,137 [p < 0.03]), a trend toward decreased length of stay (4.0+/-3.0 vs 3.4+/-2.4 days [p = 0.13]), and no significant change in the 30-day readmission rate (13% of patients vs 16% of patients [p = NS]). PMID- 9732906 TI - Serum interleukin-6 in congestive heart failure secondary to idiopathic dilated cardiomyopathy. AB - Increased serum interleukin-6 (IL-6) was associated with a higher incidence of New York Heart Association functional classes III to IV and worse left ventricular function during follow-up. Patients with elevated serum IL-6 had poor prognosis. These results reinforce the concept that increased serum IL-6 may also play an important role in disease progression. PMID- 9732907 TI - Comparison of three different atrial septal defect occlusion devices. AB - Three different devices were used to close secundum-type atrial septal defects in 28 patients. The "Amplatzer" is associated with an easier and shorter procedure than are the "Sideris Buttoned Device" and the Microvena "Angel Wings" devices. PMID- 9732908 TI - Effects of pulmonary balloon valvuloplasty on right coronary artery blood flow in pulmonary valve stenosis. AB - Pulmonary balloon valvuloplasty results in improvement in the right coronary artery blood flow velocity pattern and the volumetric flow in patients with pulmonary valve stenosis. These changes are closely related to concomitant changes in right ventricular systolic pressure. PMID- 9732909 TI - Risk of atrioventricular block during adenosine pharmacologic stress testing in heart transplant recipients. AB - Pharmacologic stress testing with adenosine in heart transplant recipients implies a high risk of atrioventricular block. Dipyridamole is preferable as a coronary vasodilator. PMID- 9732910 TI - Effect of niacin supplementation on fibrinogen levels in patients with peripheral vascular disease. AB - This study demonstrates that niacin supplementation decreases plasma fibrinogen and low-density lipoprotein cholesterol in subjects with peripheral vascular disease randomized to receive niacin, warfarin, antioxidants, or placebo. Changes in fibrinogen levels are highly correlated with changes in low-density lipoprotein cholesterol (r = 0.61; p < 0.009) in subjects taking niacin. PMID- 9732911 TI - Hemodynamic effects of arbutamine. AB - This study examined the hemodynamic effects of arbutamine, a synthetic catecholamine, in 12 patients with and 7 patients without coronary artery disease. Arbutamine produced a balanced positive inotropic (increase in left ventricular dp/dt) and chronotropic effect (increase in heart rate). PMID- 9732912 TI - Rationale and design of the Myocardial Ischemia Reduction With Aggressive Cholesterol lowering (MIRACL) Study that evaluates atorvastatin in unstable angina pectoris and in non-Q-wave acute myocardial infarction. PMID- 9732913 TI - Twenty-four more days. PMID- 9732914 TI - Echocardiography or radionuclide methods for assessment of left ventricular function in acute myocardial infarction. PMID- 9732915 TI - Nonsteroidal antiinflammatory drugs and gallstone disease: will an aspirin a day keep the gallstones away? PMID- 9732916 TI - Reversal of gastric atrophy after Helicobacter pylori eradication: is it possible or not? PMID- 9732917 TI - Has the impact of Helicobacter pylori therapy on ulcer recurrence in the United States been overstated? A meta-analysis of rigorously designed trials. AB - OBJECTIVE: The aim of this study was to assess the effect of H. pylori eradication on ulcer recurrence in North American duodenal ulcer patients by examining only treatment studies that met rigorous methodologic criteria. METHODS: Data sources were computerized bibliographic searches from 1983, review of reference lists, communication with companies that manufacture medications used for H. pylori therapy in the U.S., and H. pylori investigators, review of open presentations to the Food and Drug Administration, and review of abstracts from annual scientific meetings. Criteria for study inclusion were double blind, randomized North American trials of H. pylori therapy for duodenal ulcer, scheduled endoscopic follow-up exams for > or = 6 months, and H. pylori cure documented > or = 4 wk after completion of therapy by at least two endoscopic biopsy tests. Seven relevant trials were identified. Data were abstracted independently and disagreement was resolved by consensus. We obtained missing data and identified erroneous assessments through contact with an author or sponsor of all studies. RESULTS: The common odds ratio for ulcer recurrence was 0.20 (95% CI, 0.13-0.31) and 2.8 patients would need to be successfully treated to prevent one ulcer recurrence at 6 months. The pooled ulcer recurrence rate at 6 months in patients with H. pylori eradication was 20%. CONCLUSION: Results of North American studies of highest methodological quality confirm that H. pylori eradication markedly decreases ulcer recurrence. Nevertheless, 20% of patients in these studies had ulcer recurrence within 6 months, despite successful cure of infection and no reported use of NSAIDs. Non-H. pylori, non-NSAID ulcers may be more common in the U.S. than previously believed. PMID- 9732918 TI - Nonsteroidal antiinflammatory drug use and gallstone disease prevalence: a case control study. AB - OBJECTIVES: Conflicting results on the relationship between gallstone disease and the use of nonsteroidal antiinflammatory drugs (NSAIDs) have been reported, but studies on the effect of NSAID use in populations not selected on the basis of a high risk for gallstone development are still lacking. METHODS: We conducted a case-control study involving 216 patients, regular NSAID users (43 men and 173 women) consecutively admitted to a rheumatology department, suffering from rheumatoid arthritis (n = 147), osteoarthritis (n = 49), and ankylosing spondylitis (n = 20). Two-hundred sixteen patients who were not NSAID users, matched for gender, age, and body mass index, consecutively admitted to a medical department for various medical pathologies, acted as a control group. All patients underwent upper abdomen ultrasonography. RESULTS: The overall prevalence of gallstones was similar in the two groups: 24.0% in NSAID users (15.7% actual stones and 8.3% previous cholecystectomy) and 21.3% in controls (13.9% gallstones and 7.4% cholecystectomy). The prevalence of gallstone disease was significantly higher in women than in men, and the mean age was higher in gallstone patients than in gallstone-free patients, in both groups. No significant differences in type and duration of arthritis condition, type and dose of NSAID taken, and duration of treatment between gallstone patients and gallstone-free patients were found. On logistic regression analysis only female gender, aging, and family history of gallstone disease were significantly associated with the presence of gallstones, whereas no relationship between NSAID use and gallstone disease was found. CONCLUSIONS: Chronic NSAID ingestion does not seem to prevent gallstones in arthritis patients; in these patients gallstone disease is associated with classic risk factors (female gender and age). PMID- 9732919 TI - Reversal of fundic atrophy after eradication of Helicobacter pylori. AB - OBJECTIVES: We sought to evaluate the effect of Helicobacter pylori eradication in patients with fundic atrophic gastritis. METHODS: Acid secretion, gastric emptying, and histology were evaluated in 20 patients with fundic atrophic gastritis and H. pylori infection. After investigation, 10 patients (Group 1) received an eradicating treatment and 10 (Group 2) did not receive any treatment. One year later, the baseline investigations were repeated. Subsequently, patients in Group 2 received the same treatment given to patients in Group 1 and were reevaluated 12 months later. A further follow-up was performed in both groups 36 months after the treatment. RESULTS: At 1-yr follow-up, all the patients in Group 1 were H. pylori negative whereas all the patients in Group 2 were still infected. In Group 1, there was a significant improvement of both fundic atrophy and acid secretion, compared with baseline (p < 0.01). In Group 2, no substantial modification of either histological or functional parameters was observed at the first follow-up; conversely, a significant (p < 0.01) improvement of fundic atrophy and acid secretion was detected in these patients 12 months after eradication of the bacterium. Histological pattern remained unchanged at 36 months of follow-up in both groups. Gastric emptying remained, on the average, unaffected by the treatment; however, three patients with delayed gastric emptying at entry had normal gastric emptying after eradication of H. pylori. CONCLUSIONS: Our data suggest that mucosal atrophy can be reduced or even reversed by the eradication of H. pylori, and this is associated with a recovery of gastric function. PMID- 9732920 TI - Clarithromycin-resistant Helicobacter pylori in patients with duodenal ulcer in the United States. AB - BACKGROUND: Clarithromycin is a key component of several antimicrobial treatment regimens for Helicobacter pylori. Cure rates with clarithromycin-containing regimens are significantly decreased when resistance is present. Resistance develops by a point mutation in the ribosomal RNA of some organisms exposed to clarithromycin. We studied the prevalence of clarithromycin-resistant organisms in patients with duodenal ulcer in the United States from 1993-96. METHODS: Patients with endoscopic evidence of a duodenal ulcer were studied. Gastric biopsies were cultured for H. pylori and antimicrobial sensitivity was determined by the E-test (epsilometer agar diffusion gradient). RESULTS: In 1993-94, three of 78 patients (4%) had clarithromycin-resistant strains of H. pylori. In 1995 96, 44 of 348 patients (12.6%; p = 0.025) had resistant strains of H. pylori. Patients who had previously failed antimicrobial treatment for H. pylori accounted for much of the increase in resistant strains (25%). CONCLUSIONS: Failed therapy with clarithromycin-based regimens is a growing cause of antimicrobial resistance in H. pylori in the United States. Whereas the overall rates of primary resistance are low, the increase in secondary resistance over a short period of time is worrisome. New treatments that prevent the emergence of resistance may be important in the future. PMID- 9732921 TI - A prospective randomized trial comparing the use of omeprazole-based dual and triple therapy for eradication of Helicobacter pylori. AB - BACKGROUND: Controversy surrounds the optimal composition, dosage, and duration of therapies for eradication of Helicobacter pylori. We prospectively compared omeprazole-based dual and triple therapies in the eradication of H. pylori in a randomized manner. METHODS: Between June 1995 and March 1997, 1000 consecutive patients with acid-peptic disease associated with H. pylori infection (duodenal ulcer, 388 patients, gastric ulcer, 179 patients; duodenitis, 173 patients; gastritis, 260 patients) were prospectively recruited. They were randomized to either a 2-wk (OA) course of omeprazole 20 mg and amoxicillin 1 g, both given twice daily, or treatment for 1 wk (OCM) with omeprazole 20 mg once daily, clarithromycin 500 mg twice daily, and metronidazole 400 mg twice daily. RESULTS: The age of these 1000 patients ranged from 16 to 90 yr, with a mean of 54.9 yr. Side effects occurred in 29.6% (95% confidence interval [CI] 25.6-33.8%) and 10.6% (95% CI 8.0-13.6%) of patients taking OCM and OA, respectively (p < 0.0001). Apart from taste disturbance, however, there were no significant differences in the incidences of side effects between the two groups. One patient in the OA group and four patients of the OCM group could not tolerate the medications, and therefore did not complete treatment (p = 0.37). Seven and 13 patients in the OA and OCM groups, respectively, refused a second endoscopy (p = 0.25). The remaining 975 patients underwent a second endoscopy. Positive endoscopic findings were significantly more common in the OA group (51/492; 10.4%; 95% CI 7.8-13.4%) than in the OCM group (25/483; 5.2%; 95% CI 3.4-7.5%) in the per-protocol (PP) analysis (p = 0.004). On intent-to-treat (ITT) analysis, the overall eradication rates in the OA and OCM groups were 73.6% (95% CI 69.5 77.4%) and 92% (95% CI 89.3-94.2%), respectively (p < 0.0001). On PP analysis, the corresponding rates were 74.8% (95% CI 70.7-78.6%) and 95.2% (95% CI 92.9 97.0%), respectively (p < 0.0001). CONCLUSIONS: A course of omeprazole, clarithromycin, and metronidazole for 1 wk is a safe, well-tolerated, efficacious, and cost-effective treatment for H. pylori infection. PMID- 9732922 TI - Dyspepsia in relation to Helicobacter pylori infection and psychosocial work stress in white collar employees. AB - OBJECTIVE: We undertook an investigation of the relationship between psychosocial work stress and Helicobacter pylori (H. pylori) infection with dyspepsia. METHODS: We conducted a cross-sectional study among 189 employees of a health insurance company in the city of Ulm, Germany. RESULTS: A clear association between work-related psychosocial factors and the occurrence of dyspeptic symptoms during the past 3 months was evident. Persons who were considered to have a critical style of coping with work demands suffered more often from dyspeptic symptoms. Current infection with H. pylori was not associated with prevalence of dyspeptic symptoms. These results were also confirmed by adjustment for age, gender, smoking status, education, and use of antiinflammatory drugs within the past 3 months, by means of multivariate analysis. The odds ratio (OR) for having a dyspepsia symptom score in the upper tertile versus the 1st or 2nd was 3.22 (95% confidence interval [CI], 1.56-6.65), given that the employee was considered to have a critical style of coping with work demands. The OR for having a dyspepsia symptom score in the upper tertile given H. pylori infection was 1.23 (95% CI, 0.44-3.46), indicating no association of current H. pylori infection with dyspeptic symptoms. CONCLUSIONS: A critical style of coping with work demands may be an important determinant for dyspepsia-like symptoms. Therefore, in the absence of an underlying disease, specific intervention programs should be targeted at the behavior of the affected individual (e.g., stress-reduction programs) rather than on the treatment of specific symptoms or infection with H. pylori. PMID- 9732923 TI - Biopsy forceps disinfection technique does not influence Helicobacter pylori culture. AB - OBJECTIVES: Culturing Helicobacter pylori (Hp) has a low sensitivity rate, and is affected by factors such as the number of biopsies, transport, and culture conditions. Hp detection is also influenced by omeprazole, antibiotics, bismuth salts, or benzocaine use. Disinfection procedures based on glutaraldehyde are highly effective in eliminating any Hp contamination of endoscopic equipment. However, the possibility that some residual glutaraldehyde present in biopsy forceps after decontamination could affect Hp viability has not yet been investigated. METHODS: Antral specimens from 25 patients with active gastric or duodenal ulcer obtained with three forceps (sterilized with ethylene oxide, glutaraldehyde, or glutaraldehyde-phenolate) were streaked on appropriate media, and results of culture evaluated. RESULTS: Helicobacter pylori was isolated in 17 patients. Positivity of culture was independent of the way the forceps were sterilized, and the number of colonies (mean +/- SD) was similar for the three types of forceps (475 +/- 312, 533 +/- 242, and 550 +/- 225 colony-forming units [CFUs] for ethylene oxide, glutaraldehyde, and glutaraldehyde-phenolate, respectively). Moreover, the incubation time since isolation was also similar (6.0 +/- 1.3, 5.8 +/- 1.2, and 5.7 +/- 1.2 days for ethylene oxide, glutaraldehyde, and glutaraldehyde-phenolate disinfected forceps, respectively). CONCLUSION: The use of glutaraldehyde to sterilize biopsy forceps is not responsible for the false-negative results of Hp culture. PMID- 9732924 TI - Gastrointestinal hemorrhage in patients with systemic sclerosis and CREST syndrome. AB - OBJECTIVES: Systemic sclerosis (SSc) and calcinosis, Raynaud's phenomenon, esophageal disease, sclerodactyly, telangiectasia (CREST) syndrome present distinctive microvasculature lesions that are thought to be responsible for tissue damage and disease progression. Involvement of the gastrointestinal tract may lead to the occurrence of profuse hemorrhage. We performed a study to assess the incidence and characteristics of gastrointestinal hemorrhage in a large group of patients with SSc and CREST syndrome. METHODS: We reviewed the medical records of 144 patients with SSc/CREST seen at our institution during the period 1985 1996. Endoscopic findings and clinical data were correlated. Data are expressed as means +/- SD. RESULTS: Twenty-two of 144 (15.2%) patients had at least one episode of gastrointestinal hemorrhage (16 women, 6 men; mean age, 59.4 +/- 17.6 yr). Eight patients (8/22; 36%) had multiple episodes and four (4/22; 18%) required chronic transfusion therapy. Mucosal telangiectasias were the most common cause of bleeding (9/22; 40.9%), followed by peptic ulcer disease (7/22; 31.8%) and erosive gastritis (3/22; 13.6%). Bleeding telangiectasias occurred in the entire gastrointestinal tract, including oral cavity (n = 1), esophagus (n = 1), stomach (n = 3), duodenum (n = 1), ileum (n = 1), cecum (n = 2), and colon (n = 2). Mortality was 22.7% in patients with gastrointestinal bleeding, compared with 7.3% in patients without bleeding. CONCLUSIONS: Patients with SSc/CREST syndrome are at risk of developing severe gastrointestinal hemorrhage. This complication is associated with frequent hospitalization, blood transfusions, and increased mortality. Mucosal telangiectasias are the most common source of bleeding. Appropriate endoscopic intervention is recommended in evaluating and preventing bleeding in patients with SSc/CREST. PMID- 9732925 TI - Military history of patients with inflammatory bowel disease: an epidemiological study among U.S. veterans. AB - OBJECTIVES: The military history of patients with inflammatory bowel disease (IBD) contains types of exposure that are not available through other sources and may provide clues about the as-yet unknown etiology of IBD. We therefore sought to describe the epidemiology of IBD among veterans, with particular emphasis on their military history. METHODS: A case-control study compared 10,544 IBD patients and 42,026 controls with respect to age, gender, ethnicity, time period of military service, military duty in Vietnam, status as prisoner of war, and exposure to Agent Orange. RESULTS: Subjects with Crohn's disease were younger than those with ulcerative colitis or without IBD (odds ratio: 0.85; 95% confidence interval [CI]: 0.83-0.87). Both types of IBD affected female veterans significantly more often than male veterans, the relative female predominance being more pronounced in Crohn's disease than ulcerative colitis (0.70; 0.61-0.81 vs 0.83; 0.71-0.96). Whites were more prone to develop both types of IBD than nonwhites (2.46; 2.27-2.68 vs 2.11; 1.95-2.27). Military duty in Vietnam and a status as prisoner of war both exerted a protective influence against Crohn's disease (0.84; 0.75-0.96 and 0.60; 0.41-0.87, respectively), but not ulcerative colitis. CONCLUSIONS: The results are consistent with the hypothesis that exposure to poor sanitation decreases the future risk of developing Crohn's disease. PMID- 9732926 TI - Clinical status of ulcerative colitis in patients who smoke. AB - OBJECTIVES: Ulcerative colitis (UC) is largely a disease of nonsmokers. There are few patients who are current smokers, but we have identified a group and reviewed their clinical status, disease activity, and nicotine exposure to examine whether they remain well controlled while smoking. METHODS: Fifty-one patients from three centers with verified UC were reviewed. RESULTS: Thirty of the group were men; mean age 50 yr, with a mean age of onset of 37 yr. Twenty-two patients had proctosigmoid disease, 12 involvement of left colon, and 17 total colitis. All were current smokers; 41 were cigarette smokers averaging 17 daily. At the onset of colitis 30 were nonsmokers, 25 of them were ex-smokers and 19 developed colitis within 2 yr of stopping smoking. Twenty-eight believed smoking improved disease activity and none felt smoking had a detrimental effect on their UC. Eleven were receiving no medication for UC, 40 were receiving 5-ASA (5 aminosalicylic acid) preparations, and only two took oral steroids. All were in clinical remission, with the exception of one patient; mean St. Marks score was 1.5, out of a possible total of 22. Sigmoidoscopic grades were inactive in all patients except three. Histological assessment showed significant activity in only five. Median serum nicotine was 8 ng/ml (range, 0.4-24.4), median serum cotinine 180 ng/ml (range, 20-453), with corresponding salivary cotinine of 255 ng/ml (range, 34-683). Median rise in nicotine 2 min after a cigarette in 35 patients was 12.1 ng/ml (range, 0.4-44). CONCLUSIONS: Because most current smokers with UC have inactive disease, smoking may contribute to the clinical remission in these patients. PMID- 9732928 TI - Proliferative patterns of rectal mucosa as predictors of advanced colonic neoplasms in routinely processed rectal biopsies. AB - OBJECTIVES: We sought to determine whether the evaluation of rectal cell proliferation in routinely processed rectal biopsies of apparently normal mucosa can predict the presence of advanced colonic neoplasms. METHODS: Fifty consecutive patients, who did not meet any of the following exclusion criteria, underwent total colonoscopy. Patients with nonadvanced adenomas, inflammatory bowel disease, hereditary predisposition to colonic cancer, or a history of colonic neoplasms were excluded. Patients with neoplasms in the distal 40 cm of the large bowel were also excluded. An adenoma was considered advanced if it had a diameter > 1 cm, or villous or severe dysplasia histology were present. In 26 of the 50 patients (Group A: 16 men, 10 women; mean age, 65 yr) advanced colonic neoplasms (advanced adenomas or cancer) were detected; in the remaining 24 (Group B: 13 men, 11 women; mean age, 66 yr) the large bowel was free of neoplasms. In all patients the proliferative patterns of apparently normal rectal mucosa were evaluated using the monoclonal antibody MIB-1 to assess the expression of Ki-67 antigen in routinely processed tissues. Proliferation index for the entire crypt, as well as proliferation indices for each of the five equal compartments, into which the crypt had been divided longitudinally, were calculated for each patient. RESULTS: The mean proliferation indices were similar between the two groups compared. The mean proliferation index for the upper crypt compartments (4 + 5) in the Group A patients was significantly higher than for those of the Group B patients (p < 0.01). Multivariate stepwise logistic regression analysis revealed that among gender, age, and proliferative parameters, the pattern of cell proliferation in the upper rectal crypt (4 + 5) compartment was the only predictor of advanced colonic neoplasms (beta = 11.01, p < 0.001). CONCLUSIONS: Our data suggest that the evaluation of the upward expansion of the rectal crypt proliferative zone in routinely processed rectal biopsies of apparently normal mucosa appears to predict the presence of advanced colonic neoplasms. These preliminary results should be confirmed in larger studies. PMID- 9732927 TI - Slow-release lanreotide treatment in endocrine gastrointestinal tumors. AB - OBJECTIVES: Lanreotide is a somatostatin analogue whose activity persists for 10 14 days. In this study, we treated a group of patients with gastrointestinal endocrine tumors with lanreotide to assess its therapeutic efficacy and tolerability. METHODS: Eighteen patients, 12 male and six female, mean age 58 yr (range, 25-80 yr) were studied. Ten had carcinoid tumors, five had nonfunctioning endocrine tumors, two had glucagonomas, and the remaining one had a gastrinoma. All patients had somatostatin receptors, demonstrated by octreoscan scintigraphy. Lanreotide was administered intramuscularly at a dose of 30 mg every 10 days, for a mean of 12 months (range, 5-18 months). Fifteen of the 18 patients had been previously treated with octreotide. RESULTS: In patients with carcinoid tumors, lanreotide markedly reduced daily bowel movements and flushing episodes. A reduction was also observed in urinary serotonin and urinary 5 hydroxyindoleacetic acid, although it was not statistically significant. A marked reduction in symptoms, and in plasma glucagon and serum gastrin levels, was also observed in patients with glucagonoma and gastrinoma. In the five patients with nonfunctioning endocrine tumors, as in all the other 13 patients, no significant effects were noted in the size of the tumor. The administration of lanreotide did not cause side effects, apart from transient abdominal pain and pain at the injection site in two patients. Only in the patient with gastrinoma was lanreotide suspended, because of the appearance of attacks of marked hypoglycemia. In the 15 patients previously treated with octreotide, no differences in the effects were noted with lanreotide. CONCLUSIONS: Lanreotide has a satisfactory therapeutic efficacy and tolerability in the treatment of gastrointestinal endocrine tumors; its effects are similar to those of octreotide. However, unlike octreotide, it can be administered once every 10-14 days, instead of 2 or 3 times daily and for this reason, it is preferable in clinical practice. PMID- 9732929 TI - Comparison of two bowel preparations for colonoscopy: sodium picosulphate with magnesium citrate versus sulphate-free polyethylene glycol lavage solution. AB - OBJECTIVES: Adequate preparation of the bowel is essential for accurate colonoscopic examination. We compared colonic preparation with sodium picosulphate plus magnesium citrate (SPS-Mg) with sulphate-free polyethylene glycol electrolyte lavage (PEG-EL) solution before colonoscopy, for quality of bowel cleansing, patient discomfort, and side effects. METHODS: Sixty-eight consecutive patients were randomly assigned to receive either 3 sachets of SPS-Mg (16.5 g each) (n = 39) or 3 L of PEG-EL (n = 29) on the day before colonoscopy. Shortly before the procedure each patient was interviewed to determine the degree of discomfort (1 = none or mild, 2 = moderate, 3 = severe) and side effects. The quality of bowel cleansing was graded by a gastroenterologist who was unaware of the method of preparation (from 1 = poor to 4 = excellent). RESULTS: Of the 29 PEG-EL patients, four (14%) did not complete the preparation because of side effects. The degree of discomfort was significantly greater with PEG-EL (mean score, 2.3 +/- 0.7) than with SPS-Mg (mean score, 1.4 +/- 0.5; p < 0.01). Nausea and vomiting were significantly more common in the PEG-EL group (38% vs 13%; p < 0.05). Using intention-to-treat analysis, bowel cleansing proved to be significantly better with SPS-Mg than with PEG-EL (mean score +/- SD, 3.05 +/- 0.9 and 2.57 +/- 1.0, respectively; p = 0.036). CONCLUSIONS: Colonic preparation with SPS-Mg is better tolerated, associated with significantly fewer side effects, and results in higher quality bowel cleansing than preparation with PEG EL. PMID- 9732930 TI - Femoral neck osteopenia in patients with inflammatory bowel disease. AB - OBJECTIVE: The mechanism of bone loss in patients with inflammatory bowel disease (IBD) is not completely understood. The aim of this study was to assess indices of bone turnover and bone mineral density (BMD) in the lumbar spine and femoral neck in IBD patients. METHODS: Sixty-three patients with Crohn's disease and 41 with ulcerative colitis were studied. Serum bone-specific alkaline phosphatase (B ALP), osteocalcin, parathyroid hormone (PTH), 25 hydroxyvitamin D, interleukin-6 (IL-6), and urinary N-telopeptide cross linked type 1 collagen (NTX) were determined. BMD of the lumbar spine and femoral neck was determined by dual x-ray absorptiometry in 59 patients. RESULTS: In the femoral neck 42% of the patients had osteopenia (-2.5 SD < BMD T score < -1 SD) and another 41% had osteoporosis (BMD T score < -2.5). In the spine 34% of the patients had osteopenia and additional 42% had osteoporosis. BMD T scores were lower in the femoral neck compared to the spine. Reduced BMD was unrelated to gender, disease type, lifetime corticosteroid dose, but inversely correlated with disease duration (r = -0.36, p < 0.05). Serum IL-6 was higher in IBD patients compared to controls. A reduced level of osteocalcin, a marker of bone formation, was present in 7% of patients and an increase in NTX, a marker of bone resorption, in 25% of them. Osteoporotic IBD patients (spine or hip BMD T score < -2.5) had increased serum IL-6, osteocalcin and PTH level compared to nonosteoporotic patients. CONCLUSIONS: There is a high prevalence of reduced BMD at the spine and femoral neck in IBD patients, which is more severe in the hip. Bone turnover in osteoporotic IBD patients is associated with an increase in osteocalcin, PTH and IL-6. IL-6 may play a role in the pathogenesis of bone loss in IBD. PMID- 9732931 TI - Outcome states of colorectal cancer: identification and description using patient focus groups. AB - OBJECTIVE: Utilities for the outcome states of colorectal cancer must be measured to evaluate the cost-utility of screening and surveillance strategies for this disease. We sought to identify these outcome states, define their associated areas of morbidity, and construct representative descriptions of them for use in a utilities assessment instrument. METHODS: We identified candidate colorectal cancer outcome states based on a review of the literature and interviews with health care professionals. We organized patient focus groups from each of the candidate outcome states to examine their homogeneity and define their associated areas of morbidity. After analyzing the focus group transcripts, we identified and described outcome states of colorectal cancer for future incorporation into a utilities assessment instrument. RESULTS: Six candidate outcome states of colorectal cancer were identified based on disease stage and location at diagnosis. Thirty-eight patients then participated in six focus groups. Analysis of the focus group transcripts revealed seven areas of morbidity associated with colorectal cancer. These areas included problems with social interaction and cognition, fear of cancer recurrence, pain, fatigue, changes in bowel habits, and sexual dysfunction. Based on differences in the intensity and frequency of the symptoms reported in each of these areas, seven distinct outcome states of colorectal cancer were identified and described. CONCLUSION: Clinically distinct outcome states of colorectal cancer are determined by the stage and location of the cancer at the time of diagnosis. Descriptions of these outcome states were created using data collected from patient focus groups. These descriptions can be incorporated into a utilities assessment instrument. PMID- 9732932 TI - Relationship between biliary and serum bile acids and response to ursodeoxycholic acid in patients with primary biliary cirrhosis. AB - OBJECTIVE: Ursodeoxycholic acid (UDCA) improves liver biochemistries and enriches the bile with UDCA in patients with primary biliary cirrhosis. The aim of this study was to determine whether the degree of enrichment of bile correlated with that of serum and whether either of these measures correlated with improvement in measures of liver disease. METHODS: In a randomized study, biliary and serum bile acid analyses were performed at entry and after 2 yr of UDCA or placebo. RESULTS: The percentage of ursodeoxycholic acid in bile increased by 42% in the UDCA group (n = 61) compared with 8% in the placebo group (n = 57) (p < 0.0001). Measurement of serum bile acids in 32 patients (18 ursodeoxycholic acid, 14 placebo) indicated that at 2 yr, ursodeoxycholic acid comprised 65% of serum bile acids in the treated group and 7% in the placebo group. Agreement between bile and serum was fair (r = 0.75, p < or = 0.00002) because in some patients, plasma but not biliary bile acids were enriched with UDCA. Changes in biliary ursodeoxycholic acid correlated significantly but weakly with the changes in serum alkaline phosphatase, AST, bilirubin, and in Mayo risk score. Correlations between changes in serum bile acid composition and biochemical measures of disease activity were even weaker. CONCLUSION: The measurement of biliary bile acids is superior to that of serum bile acids for assessing the compliance and changes in the circulating bile acids in patients receiving ursodeoxycholic acid for the treatment of primary biliary cirrhosis. Furthermore, measures to further increase the proportion of ursodeoxycholic acid in circulating bile acids should be explored. PMID- 9732933 TI - Serum apolipoprotein(a) concentrations and Apo(a) phenotypes in patients with liver cirrhosis. AB - OBJECTIVE: The liver is the major site of apolipoprotein(a) synthesis, and an inverse correlation between the size of apolipoprotein(a) isoforms and its serum levels have been described. We evaluated the Apo(a) serum levels and its isoforms in patients with liver cirrhosis at different stages of the disease (Childe Turcotte classification), and during the characteristic phase of liver synthesis decline. METHODS: We studied 84 patients with liver cirrhosis and 185 control subjects with normal liver function. RESULTS: Apo(a) serum levels were significantly lower (p < 0.01) in cirrhotic patients and, after 24 months, six patients showing a change from class A to class B had a statistically significant decrease in Apo(a) concentrations (p = 0.0313). Moreover, our data showed an inversion of the small/large isoforms ratio in patient with cirrhosis in spite of the reduction in plasma concentration. CONCLUSION: We showed a reduction of Apo(a) serum concentrations in a large number of patients with cirrhosis and, for the first time, during the characteristic phase of liver synthesis decline, confirming the liver as the major site of Apolipoprotein(a) synthesis. Moreover we showed in the cirrhotic patients that the normal correlation between Apo(a) isoforms and Apo(a) concentrations is not conserved and the low levels are not dependent upon a high prevalence of large isoforms. PMID- 9732934 TI - Unexpected clinical remission of cholestasis after rifampicin therapy in patients with normal or slightly increased levels of gamma-glutamyl transpeptidase. AB - OBJECTIVE: Rifampicin is an effective drug against pruritus in intrahepatic cholestasis. However, there is no specific hepatic disease in which its use could cause undoubtedly biochemical improvement. The aim of this study was to describe patients with complete remission of cholestatic symptoms after rifampicin therapy. METHODS: We reported three female patients with intrahepatic cholestasis with no evidence of viral, metabolic, or autoimmune liver diseases. Total bilirubin levels ranged from 13.2 to 27.2 mg/dl (before the first treatment with rifampicin), and in all of them gamma-glutamyl transpeptidase values were within the normal range or slightly increased. Rifampicin therapy was administered orally, without any concomitant drug, with an effective dosage of 5-17 mg/kg/day. RESULTS: In all patients, pruritus ceased completely and bilirubin returned to normal values. The symptoms recurred after rifampicin withdrawal on, at least, three occasions in each patient, and these symptoms were always eliminated after its reintroduction. The patients had a total of 16 cholestatic episodes during a follow-up of 8 yr, with a complete clinical recovery in all of them. Undergoing therapy with a suitable dosage of rifampicin, none of the patients had a cholestatic crisis even during a period for as long as 12 months. The diagnosis of two patients was consistent with benign recurrent intrahepatic cholestasis, and it was not well defined in the remaining. CONCLUSION: Rifampicin may induce clinical remission, and perhaps prevent clinical relapses of intrahepatic cholestasis with normal or slightly increased levels of gamma-glutamyl transpeptidase. PMID- 9732935 TI - Increased acylCoA-cholesterol ester acyltransferase activity in gallbladder mucosa in patients with gallbladder cholesterolosis. AB - OBJECTIVE: Although cholesterolosis of the human gallbladder is a relatively common disease, its etiology has not been fully understood. The aim of this study was to determine this etiology. METHODS: The lipid composition of the gallbladder mucosa and gallbladder bile and the enzyme activities (acylCoA-cholesterol ester acyltransferase [ACAT] and cholesterol ester hydrolase [CEH]) of the gallbladder mucosa were measured in control subjects, patients with cholesterolosis, and patients with cholesterol gallstone disease. RESULTS: Levels of cholesterol ester in gallbladder mucosa in patients with cholesterolosis (n = 12) were higher than those in control subjects (n = 8). With regard to the lipid content in gallbladder bile, no differences were found in concentrations of cholesterol, phospholipids, and bile acids among control subjects (n = 11), patients with cholesterolosis (n = 13), and those with cholesterol gallstone disease (n = 15). In gallbladder mucosa, ACAT activity was significantly higher in patients with cholesterolosis (n = 10) than in control subjects (n = 8), whereas CEH activity did not differ between the two groups. As a result, the ACAT/CEH activity ratio was higher in patients with cholesterolosis than in control subjects. CONCLUSIONS: It would be suggested that cholesterol ester synthesis of gallbladder mucosa might play an etiological role in the development of cholesterolosis. PMID- 9732936 TI - No evidence for overexpression of the p53 protein and mutations in exons 4-9 of the p53 gene in a large family with adenomatous polyposis. AB - OBJECTIVE: Familial adenomatous polyposis coli (FAP) is an autosomal dominant disease characterized by an early onset of numerous adenomatous polyps of the colon and a high risk of colon carcinoma. The role of the p53 gene in the multistage process of FAP is as yet poorly defined. In the present study, a large family with evidence of polyposis and colon cancer was screened for the mutations of the p53 gene and protein overexpression. METHODS: We examined p53 protein expression from individuals with immunohistochemical techniques using monoclonal antibody PAb1801. Polymerase chain reaction products of exons 4-9 of the p53 were examined from individuals by single strand, conformational polymorphism analysis. RESULTS: We could find no evidence of overexpression and mutations of the p53 in any lesion including adenomas and carcinomas. CONCLUSION: We found that p53 gene alterations do not contribute to the genesis of adenoma or carcinoma of FAP patients for this large family examined. PMID- 9732937 TI - Gluten-free diet induces regression of T-cell activation in the rectal mucosa of patients with celiac disease. AB - OBJECTIVE: An increase in the number of intraepithelial lymphocytes (IEL) in the rectal epithelium of patients with active celiac disease has been described. No data are available about how they vary during a gluten-free diet. The aim of the study was to assess the effect of a gluten-free diet on T-cell activation in the rectal mucosa of adult patients with celiac disease. METHODS: Frozen duodenal and rectal biopsies were available in four celiac patients (one male, three female, mean age 39 yr) both before and after 7 to 24 months on a gluten-free diet. Biopsy samples were stained using monoclonal antibodies directed against CD3, betaF1, TcRdelta1, CD25, and HLADR. Numbers of IEL were estimated by counting the peroxidase-stained cells per 100 epithelial cells. Four patients without histological abnormalities were used as control subjects. RESULTS: In the four patients with active celiac disease but in none of the controls, CD25 was expressed by both duodenal and rectal lamina propria cells and HLADR was expressed by duodenal (4/4) and rectal (2/4) epithelial cells. In addition, two patients with active celiac disease had features of lymphocytic colitis, i.e., >20 IEL per 100 epithelial cells. After a gluten-free diet, the mean number of rectal CD3+ betaF1+ IEL decreased (9% vs 21%) and the expression of CD25 and HLADR was no longer present. These changes mirrored those found in the small intestinal biopsies. CONCLUSION: These results suggest that in celiac disease, gluten-driven T-cell activation is not restricted to the proximal part of the intestine but is present on the whole intestinal length. Assessment of the effectiveness of a gluten-free diet through rectal biopsies warrants investigation, as it could lessen discomfort for patients and prove more cost effective. PMID- 9732938 TI - Dual therapy using a double dose of lansoprazole with amoxicillin versus triple therapy using a double dose of lansoprazole, amoxicillin, and clarithromycin to eradicate Helicobacter pylori infection: results of a prospective randomized open study. AB - OBJECTIVES: The eradication of Helicobacter pylori is recommended in duodenal ulcer disease. The aim of this randomized open trial was to evaluate and compare H. pylori eradication and safety after a dual therapy consisting of lansoprazole (30 mg b.i.d.) and amoxicillin (1 g b.i.d.) versus a triple therapy consisting of lansoprazole (30 mg b.i.d.), amoxicillin (1 g b.i.d.), and clarithromycin (500 mg b.i.d.) administered from day 1 to day 14. METHODS: All patients with an ulcer received lansoprazole (30 mg) from day 15 to day 28. H. pylori status was determined from antral biopsies using histology, culture, and polymerase chain reaction (PCR) upon inclusion and 1-3 months after the end of the treatment. RESULTS: Of the 50 patients included in the study, five did not adhere to the protocol. H. pylori eradication was obtained in 37.5% of the patients receiving lansoprazole-amoxicillin (n = 9/24) and in 95.2% of the patients receiving lansoprazole-amoxicillin-clarithromycin (n = 20/21, p < 0.0002). Minor side effects appeared in 8.3% of the cases during dual therapy (n = 2/24) and in 52% during triple therapy (n = 13/22, p < 0.001). These side effects consisted mainly of diarrhea and a metallic taste. CONCLUSION: Concomitant administration of double doses of lansoprazole with amoxicillin and clarithromycin is very efficacious against H. pylori infection compared with dual therapy. PMID- 9732939 TI - Pseudoaneurysm of the cystic artery: a rare cause of hemobilia. AB - Aneurysms are a rare cause of hemobilia, and usually involved are branches of the hepatic and gastro-duodenal arteries. A case of a patient with hemobilia secondary to a pseudoaneurysm of the cystic artery is presented. Fewer than 10 cases have been reported in the literature, and in all of them the condition was associated with inflammation of the gall bladder, as in our case. Selective hepatic angiography is the procedure of choice for diagnosis. Upper gastrointestinal endoscopy with side-viewing scopy can demonstrate bleeding from papilla. Color-Doppler ultrasonography also may prove to be useful in equivocal cases. Cholecystectomy and ligation of cystic artery with proximal control of the hepatic artery was done at laparotomy after diagnosis was made. PMID- 9732940 TI - Inflammatory pseudotumor of the liver--a rare entity and a diagnostic challenge. AB - Inflammatory pseudotumors are rare benign lesions that occur throughout the body. Hepatic pseudotumors are uncommon lesions, accompanied by fever, malaise, abdominal pain, and mass effect, and therefore are commonly misdiagnosed as malignant tumors or liver abscesses. Because even routine imaging procedures usually fail to differentiate hepatic pseudotumors from liver neoplasms, a diagnostic histology procedure is usually needed. We present a case of hepatic pseudotumor that resolved spontaneously and review the literature, including 40 previously reported cases and the differential diagnosis. PMID- 9732942 TI - Fluoroscopy-induced radiodermatitis after transjugular intrahepatic portosystemic shunt. AB - Transjugular intrahepatic portosystemic shunt is a nonsurgical procedure used to manage the complications of portal hypertension. This report describes three cases of fluoroscopy-induced radiodermatitis after transjugular intrahepatic portosystemic shunt and reviews the characteristics and treatment of radiation induced skin reactions. PMID- 9732941 TI - Enhanced phenotypic expression of alpha-1-antitrypsin deficiency in an MZ heterozygote with chronic hepatitis C. AB - A middle-aged white man of Scotch-Irish ancestry, being treated for chronic hepatitis C, was found to be heterozygous for alpha1-antitrypsin deficiency (PiMZ phenotype) after diagnostic PAS-positive, diastase-resistant globules were detected in a liver biopsy. The globules had not been present in a biopsy obtained 4 yr previously. He was also found to be heterozygous for the cys282tyr mutation of the HFE gene, which is the chief cause of HLA-linked hereditary hemochromatosis (HHC). His liver disease progressed over 4 yr from mild hepatitis to moderate hepatitis with cirrhosis despite therapy with interferon-alpha, and phlebotomy plus interferon. These conditions appeared to have synergistic effects, with the chronic viral hepatitis unmasking the alpha1AT deficiency, and the alpha1AT deficiency (and possibly the heterozygosity for HHC), exacerbating the course of the hepatitis C. PMID- 9732943 TI - A case of glycogen storage disease type Ia with multiple hepatic adenomas and G727T mutation in the glucose-6-phosphatase gene, and a comparison with other mutations previously reported. AB - We report a case of 23-yr-old man with glycogen storage disease (GSD) type Ia complicated by multiple hepatic adenomas. Analysis of the G-6-Pase gene using peripheral blood sample showed this patient to be homozygous for a G-to-T transversion at nucleotide 727 in exon 5. This mutation is prevalent among Japanese patients, suggesting that specific genotypes may correlate with different clinical courses or outcomes. PMID- 9732944 TI - Inverted Meckel's diverticulum: an entity simulating an ileal polyp. AB - A case of inverted Meckel's diverticulum is described. This presented as an ileal polyp in an individual with chronic unexplained iron deficiency anemia. Most prolapsed Meckel's diverticula occur acutely as intussusceptions with bowel obstruction and characteristically develop in childhood. This case therefore represents an unusual surgical problem in an older individual in which the diagnosis was clinically unexpected. PMID- 9732945 TI - Spontaneous bacterial peritonitis caused by infection with Listeria monocytogenes: a case report and review of the literature. AB - Spontaneous bacterial peritonitis is a frequent and often serious complication of long-standing ascites in the presence of advanced liver disease. Coliform bacteria account for the infection in most cases and are thought to be related to translocation of bacteria from the bowel into the peritoneal cavity. The empiric use of cefotaxime is well established as most of the causative organisms are sensitive to this antibiotic. However, we report on a case of spontaneous bacterial peritonitis in a patient with hepatitis C related cirrhosis who was awaiting liver transplantation caused by infection with Listeria monocytogenes, in which the patient did not improve with empiric antibiotic therapy. This case adds to the 23 others reported in the literature since 1966. Our case raises some concerns about the universal empiric usage of cefotaxime for spontaneous bacterial peritonitis because it does not offer adequate coverage against organisms such as Listeria, enterococci, Pasturella, and anaerobes. PMID- 9732946 TI - Insulinoma associated with liver lesions: value of MR imaging. AB - We report on a middle-aged woman who presented with clinical and biochemical findings of insulinoma. Preoperative evaluation by ultrasound, CT, and angiography located the pancreatic lesion but also revealed two focal liver lesions. The latter were interpreted as metastases. MR imaging with injection of superparamagnetic iron oxide particles not only localized the insulinoma but proved to be the only noninvasive technique capable to exclude presence of liver metastases preoperatively. This reversed management to minimal laparoscopic surgery. Recent literature of preoperative imaging evaluation of insulinoma and focal liver lesions is discussed. PMID- 9732947 TI - Ibuprofen-induced hepatotoxicity in patients with chronic hepatitis C: a case series. AB - Hepatitis C is a common chronic infection. Nonsteroidal anti-inflammatory drugs are commonly ingested both over-the-counter and by prescription. This case report describes three cases where ibuprofen use leads to a marked rise in hepatic transaminases with one case repeating on rechallenge. These cases support the recommendation of acetaminophen over nonsteroidal antiinflammatory drug use in patients with chronic hepatitis C. PMID- 9732948 TI - Malignant change in a duodenal adenoma in familial adenomatous polyposis: report of a case. AB - Patients with FAP (familial adenomatous polyposis) are known to be at high risk for duodenal cancer. Although the adenoma-carcinoma sequence is thought to exist in the duodenum, clinical observation of the development of duodenal adenoma to cancer has rarely been reported. We outline a 44-yr-old postcolectomy man with FAP who underwent regular gastroduodenoscopy annually or biannually and was found to be harboring duodenal ampullary cancer 5 yr after colectomy. Endoscopic and pathological examination of the ampullary lesion during these 5 yr revealed progression of pathology from adenoma to carcinoma. Pathology of the surgical specimen confirmed ampullary cancer. This in vivo demonstration of the adenoma carcinoma sequence highlights the current limitations of duodenal surveillance in FAP. PMID- 9732949 TI - Hereditary hemorrhagic telangiectasia causing high output cardiac failure: treatment with transcatheter embolization. AB - We report a case of hereditary hemorrhagic telangiectasia complicated by high output heart failure caused by intrahepatic arteriovenous malformations. This patient was treated using transcatheter embolization of the intrahepatic arteriovenous malformations with concurrent measurement of cardiac output to monitor progress of the embolization. PMID- 9732950 TI - The Muir-Torre syndrome: a rare variant of hereditary nonpolyposis colorectal cancer associated with hMSH2 mutation. AB - The Muir-Torre syndrome is a rare autosomal dominant disorder characterized by the association of visceral malignancies with typical skin lesions. This syndrome is now considered a subtype of the more common hereditary nonpolyposis colorectal cancer syndrome (HNPCC). This last condition has been ascribed to mutations in four mismatch repair genes, and similar mutations, mostly located at hMSH2 gene, are now being described in some Muir-Torre patients. We describe the case of a 64 yr-old woman with no family history of colorectal cancer, who developed two visceral malignancies belonging to the usual spectrum of hereditary nonpolyposis colorectal cancer (colon and stomach), beginning at age 41. She additionally developed several skin tumors, including multiple keratoacanthomas, thus fulfilling Muir-Torre diagnostic criteria. Because of her cutaneous phenotype, she was screened for DNA mismatch repair gene mutations by in vitro synthetized protein assay (IVSP) and a truncating mutation was identified at hMSH2. We further discuss the clinical significance of the Muir-Torre phenotype, the association of this syndrome with hMSH2 mutations and the important implications of genetic diagnosis for the patient and her offspring. PMID- 9732952 TI - PCR analysis of T. whippelii DNA in a case of Whipple's disease: effect of antibiotics and correlation with histology. AB - A 58-yr-old man developed severe weight loss, arthralgias, and diarrhea. Endoscopic examination of the stomach and duodenum revealed thickened folds of duodenal mucosa. Biopsy of the gastric mucosa was negative, whereas duodenal biopsy revealed blunted epithelial villi and PAS-positive foamy macrophages within the lamina propria. Bacilli typical of those associated with Whipple's disease were found by electron microscopy. The diagnosis was confirmed by polymerase chain reaction (PCR) assay, which detected a portion of the 16S ribosomal RNA gene sequence corresponding to the Whipple bacillus (Tropheryma whippelii) in duodenum, stomach, and liver biopsies before therapy. T. whippelii DNA was eliminated from all tissues tested within 3 months of starting antibiotic treatment, but the histological improvement lagged behind the clinical and molecular evidence of improvement. PMID- 9732953 TI - Omeprazole for bleeding PUD: do we finally have evidence for effective medical therapy? PMID- 9732951 TI - Multiple early gastric stump carcinomas after gastrectomy for peptic ulcer. AB - The remnant stomach after partial gastrectomy is considered to have a predilection for the development of primary gastric carcinoma. However, early gastric stump carcinomas are uncommon because the diagnosis of gastric stump carcinoma is more difficult than that of carcinoma in the intact stomach. Triple early gastric stump carcinomas, as in the present case, are exceedingly rare and may provide some clues for further investigation of carcinogenesis in the gastric stump. We studied about the histological appearance, genetic alterations (P-53 gene, c-erbB-2 gene and K-ras gene), and expression of tumor-associated antigens (carcinoembryonic antigen, carbohydrate antigen 19-9, and sialyl-Tn) in this rare case. The three carcinomas differed from each other histologically. With respect to genetic alterations, c-erbB-2 was amplified in one lesion, but no mutations of K-ras and P-53 gene were detected. The three carcinomas also differed from each other on the expression of tumor-associated antigens. In noncancerous mucosal epithelium at the anastomosis showing hyperplasia and cystic formation of glandular epithelial cells, no genetic alterations were detected, but sialyl-Tn and carbohydrate antigen 19-9 were expressed. These results suggest that there may be different processes of carcinogenesis of the three carcinomas even though they occurred under identical environmental conditions to those that have increased cancer risk. PMID- 9732954 TI - For whom the bell tolls. PMID- 9732955 TI - Same-day discharge for duodenal ulcer-related bleeding--be safe or sorry? PMID- 9732956 TI - Gastric xanthelasma in a patient after partial gastrectomy. AB - Gastric xanthelasma is a benign condition that must be differentiated from early carcinoma. The first descriptions of this condition appeared in the literature >100 yr ago but, to date, no adequate photographs of the gross endoscopic appearance have been published. With the hope of increasing awareness of this condition, we report a case of gastric xanthelasma in a patient with previous partial gastrectomy and review the existing literature. We also present photographs with the typical endoscopic characteristics of the lesion. PMID- 9732957 TI - Non-Hodgkin's lymphoma presenting nine years after ileal-pouch ileoanal anastomosis for ulcerative colitis. PMID- 9732958 TI - Giant pseudopolyps presenting as colocolic intussusception in Crohn's colitis. AB - Bowel obstruction is a well-known complication of Crohn's disease and is usually a result of stricture formation. Intussusception due to giant pseudopolyps is a rare form of bowel obstruction even in Crohn's disease. These giant pseudopolyps rarely regress with medical management alone and often require surgical resection. PMID- 9732959 TI - Primary intestinal cryptococcosis mimicking adenomatous polyp in an HIV-negative patient. AB - Primary cryptococcal infection is thought to arise in the lungs, whereas secondary lesions may be found anywhere in the body. Because intestinal involvement is rare, especially in nonimmunocompromised patients, little is known about this localization. Nevertheless, the intestinal tract has long been suggested a possible portal of entry of Cryptococcus neoformans, although the hypothesis has never been sufficiently documented. We report an isolated cryptococcosis of the sigmoid colon mimicking an adenomatous polyp. The lesion has an endoscopic interest, being the first of its kind reported in the literature, and a more important pathogenic interest, as it highlights a further pathway of cryptococcal infection, one of major importance in immunocompromised patients. PMID- 9732960 TI - Treatment of chronic post-radiation proctitis with oral administration of sucralfate. AB - Several nonsurgical approaches to the treatment of postradiation proctitis have been described, but no effective conservative treatment has yet been established. As an alternative to the usual treatment, three cases of chronic postradiation proctitis with hemorrhage were successfully treated with oral administration of sucralfate, with resultant decreased bleeding in long term follow-up period. Oral sucralfate may provide a novel approach to the treatment of intractable postradiation proctitis. PMID- 9732961 TI - A simple technique to remove migrated esophageal stents. AB - A 51-yr-old man with a tracheoesophageal fistula from an esophageal carcinoma had two expandable covered stents placed, which migrated distally. After several unsuccessful attempts to remove the stents, we fashioned a homemade snare to entrap and remove the stents under endoscopic and fluoroscopic guidance. PMID- 9732962 TI - H. pylori: is it really such a bad guy after all? PMID- 9732963 TI - Body composition and energy metabolism in celiac disease. PMID- 9732964 TI - Hepatitis C virus infection related to anabolic-androgenic steroid injection in a recreational weight lifter. PMID- 9732965 TI - Diagnosis of cirrhosis and portal hypertension. PMID- 9732966 TI - Hepatitis G virus (HGV) and antibodies to a putative HGV envelope protein in alcoholic patients in southeastern France. PMID- 9732967 TI - Retrograde esophageal contraction? A late sequela to sclerotherapy. PMID- 9732968 TI - Generalized cutaneous metastases from carcinoma stomach. PMID- 9732969 TI - Ampullary carcinoid. PMID- 9732971 TI - Monitoring of unbound digoxin in patients treated with anti-digoxin antigen binding fragments: a model for the future? PMID- 9732970 TI - HCV genotypes and breakthrough in patients treated with recombinant alpha interferon. PMID- 9732972 TI - Erythrocyte folate analysis: a cause for concern? AB - Neural tube defects can be prevented by adequate intake of periconceptional folate, and inverse associations between folate status and cardiovascular disease and various cancers have been noted. Thus, there is renewed interest in the analysis of red cell folate (RCF) as an indicator of folate deficiency risk. Assessment of the assumptions that underpin RCF assays indicates that many are false. Published literature suggests that increased deoxy-hemoglobin (which can bind RCF electrostatically) yields more assayable folate, and increased oxy hemoglobin (which cannot bind RCF) yields less assayable folate. It is argued that as deoxy-hemoglobin picks up oxygen and switches quaternary structure, any bound folate must, on purely theoretical grounds, become physically "trapped". Venous blood taken for analysis is 65% to 75% saturated with oxygen, and pro-rata "trapping" will lead to serious underestimation of RCF. Hence, doubt is cast over the validity of all previous RCF values. Some strategies for accurately assessing RCF are suggested. PMID- 9732973 TI - 5-Aminolevulinic acid dehydratase deficiency porphyria: a twenty-year clinical and biochemical follow-up. AB - 5-Aminolevulinic acid dehydratase (ALAD) activity in two patients with compound heterozygous 5-aminolevulinic acid dehydratase deficiency porphyria was studied over the last 20 years. The patients' enzyme activity was <10% from 1977 to 1997. An acute crisis in each patient was successfully treated by infusion of glucose and heme arginate. After this therapy both urinary 5-aminolevulinic acid (ALA) and total porphyrins were diminished to 65% in patient B. In patient H, ALA was decreased to 80%, and total porphyrins were reduced to 15% after treatment with heme arginate and glucose. The patients remained free of symptoms after this therapy. Family studies of patient B showed cross-reactive immunological material (CRIM), in which the maternal mutation is CRIM(+), whereas the paternal mutation is CRIM(-). Incubation of erythrocyte lysates with ALA decreased porphyrin formation, whereas incubation with porphobilinogen produced porphyrin concentrations within reference values in both patients, confirming that ALAD activity is rate-limiting in these cells. PMID- 9732974 TI - Biochemical hallmarks of tyrosine hydroxylase deficiency. AB - We report the biochemical hallmarks of tyrosine hydroxylase deficiency with emphasis on reliable diagnostic strategies of four new cases of an inborn error of tyrosine hydroxylase (TH). Three of our patients from different parts of the Netherlands were found homozygous for a mutation in exon 6 (G698A) of the TH gene, and one patient was found compound heterozygous for the same mutation and an additional mutation in exon 3. The first clinical symptoms of hypokinesia, rigidity of arms and legs and axial hypotonia, developed between 3 and 7 months of age. Cerebrospinal fluid investigations revealed a characteristic metabolite constellation in every case: low homovanillic acid (HVA) and 3-methoxy-4 hydroxyphenylethyleneglycol concentrations in the presence of normal reference range 5-hydroxyindolacetic acid concentrations. Strict adherence to a standardized lumbar puncture protocol and adequate age-related reference values are essential for diagnosis of this "new" treatable neurometabolic disorder. Urinary measurements of HVA, vanillylmandelic acid, and catecholamines can lead to false-negative conclusions. All patients showed a remarkable clinical improvement on a low dose of L-dihydroxyphenylalanine/ (S)-2-(3,4 dihydroxybenzyl)-2-hydrazinpropionic acid. During treatment, cerebrospinal fluid HVA, and 3-methoxy-4-hydroxy-phenylethyleneglycol increased substantially. PMID- 9732975 TI - Capillary electrophoresis for rapid profiling of organic acidurias. AB - Organic acids analysis is a powerful technique in the diagnosis of inborn errors of metabolism. Clinically, patients present with severe symptoms, and early detection and appropriate treatment are often lifesaving. Most of the existing methods are based on gas chromatography in combination with mass spectrometry and require sophisticated equipment and complex sample pretreatment and derivatization. We propose a rapid, simple, and automated capillary electrophoretic method for routine analysis of urine to detect 27 organic acids related to metabolic diseases. With this method, direct measurements are performed on samples after initial centrifugation and dilution, if needed. Separation is performed in pH 6.0 phosphate buffer with methanol added as an organic modifier, -10 kV applied potential, and ultraviolet detection at 200 nm. The assay is completed in <15 min, and alternative separation conditions are proposed in case of overlapping peaks. The developed method allows the identification and quantitation of methylmalonic, pyroglutamic, and glutaric acids in samples of patients with diseases related to these acids. PMID- 9732976 TI - Different intracellular compartmentations of cardiac troponins and myosin heavy chains: a causal connection to their different early release after myocardial damage. AB - We investigated the net myocardial release of creatine kinase isoenzyme MB (CKMB), myoglobin, cardiac troponin T (cTnT), cardiac troponin I (cTnI), and cardiac beta-type myosin heavy chain (beta-MHC) into the coronary circulation after cardioplegic cardiac arrest in humans. Cardiac markers were measured in paired arterial, central venous, and coronary sinus blood in 19 patients undergoing elective coronary artery bypass grafting (CABG) before aortic cross clamping and 1, 5, 10, and 20 min after aortic declamping. cTnT and cTnI were released into the coronary sinus in parallel to each other and almost simultaneously to myoglobin and CKMB within 20 min of reperfusion. In contrast, no beta-MHC was released in the same patients during the study period. The average soluble cTnT and cTnI pools in right atrial appendages of 11 patients with right atrial and right ventricular pressures within reference values were comparable and were approximately 8% of total myocardial troponin content. The soluble beta-MHC pool was <0.1% in all patients. Our results demonstrate the impact of the different intracellular compartmention of regulatory and contractile proteins on their early release from damaged myocardium. PMID- 9732977 TI - Cardiac troponin T isoforms expressed in renal diseased skeletal muscle will not cause false-positive results by the second generation cardiac troponin T assay by Boehringer Mannheim. AB - The purpose of this study was to determine whether the two monoclonal anti cardiac troponin T (cTnT) antibodies (MAbs) used in the second generation cTnT assay by Boehringer Mannheim (BM, capture Ab, M11.7; detection Ab, M7) would detect cTnT isoforms expressed in human skeletal muscle in response to chronic renal disease (CRD). cTnT expression was examined in skeletal muscle biopsies obtained from 45 CRD patients, as well as nondiseased human heart (n = 3) and skeletal muscle (n = 3). cTnT proteins were resolved by modified 7.5% sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transferred to nitrocellulose, and probed with the following anti-cTnT MAbs: M11.7; M7; JS-2, Lakeland Biomedical; and 13-11, Duke University. All four antibodies detected the cTnT isoforms (Ta, Te) expressed in human myocardium. In 20 of 45 skeletal muscle biopsies, MAb M11.7 recognized its epitope in one to three proteins, molecular mass 34-36 kDa, designated Te, Td, and Tc; the strongest signal was that of Te. The same proteins were recognized by MAbs JS-2 and 13-11. The BM M7 antibody did not detect the cTnT isoforms in the molecular mass range of 34-36 kDa. However, MAb M7 did detect a cTnT isoform, molecular mass 39 kDa, in 2 of 45 biopsies. This isoform had an electrophoretic mobility similar to the predominant heart cTnT isoform, Ta. We conclude that cTnT isoforms are expressed in the skeletal muscle of CRD patients. However, given the epitopes recognized by the BM MAbs M7 and M11.7 and the variable presence of these cTnT isoforms in skeletal muscle, the second generation BM cTnT assay will not detect these isoforms if they are released from skeletal muscle into the circulation. PMID- 9732978 TI - Analytical performance and clinical application of a new rapid bedside assay for the detection of serum cardiac troponin I. AB - Detection of cardiac troponin I (cTnI) in patients suspected of having an acute coronary syndrome is highly predictive for an adverse outcome. We evaluated a bedside test for cTnI that uses a polyclonal capture antibody and two monoclonal indicator antibodies. Clinical studies were performed in patients with acute coronary syndrome and patients with chest pain but no evidence of acute myocardial injury. The whole-blood, 15-minute assay had a concordance of 98.9% with an ELISA for cTnI and a detection limit of 0.14 microg/L, and the device tolerated temperatures between 4 degrees C and 37 degrees C. Diagnostic sensitivity for myocardial infarction at arrival (3.5 +/- 2.7 h after onset of symptoms) was 60% [creatine kinase isoenzyme MB (CK-MB) mass, 48%; CK activity, 36%; P < 0.01], and 4 h later, diagnostic sensitivity was 98% (CK-MB mass, 91%; CK activity, 61%; P < 0.01). In 38% of the patients with unstable angina, at least one positive cTnI test was found (CK-MB mass, 4%; CK activity, 2%). No false-positive test results were found in renal failure or injury of skeletal muscle. We conclude that the diagnostic efficacy of the cTnI rapid test was comparable with the cTnI ELISA and superior to CK-MB determination. Therefore, this device could facilitate decision-making in patients with chest pain at the point of care. PMID- 9732979 TI - Inhibition of LDL oxidation by melatonin requires supraphysiologic concentrations. AB - Melatonin has been suggested as a potent antioxidant that may protect against development of atherosclerosis and cancer; however, these effects are unproven and controversial. The antioxidant capacity of melatonin was tested in comparison with alpha-tocopherol, ascorbic acid, and the melatonin precursors tryptophan and serotonin, by measuring inhibition of metal ion-mediated and human macrophage mediated oxidation of LDL. Melatonin had weak antioxidant activity that was detectable only at concentrations 10000- to 100000-fold higher than physiologic concentrations. These results were comparable with published data showing that the radical scavenging activity of melatonin requires markedly supraphysiologic concentrations. In contrast, alpha-tocopherol was 50- to 100-fold more potent and was efficacious at physiologic concentrations. Ascorbic acid and tryptophan also were active at physiologic concentrations and were significantly more potent than melatonin. In summary, extremely supraphysiologic concentrations of melatonin had only weak antioxidant activity, which was surpassed by alpha-tocopherol, ascorbic acid, and tryptophan. PMID- 9732980 TI - Simple method for the routine determination of betaine and N,N-dimethylglycine in blood and urine. AB - A simple and convenient method using commercially available derivatization reagents is described for the measurement of betaine and N,N-dimethylglycine (DMG) in blood and urine. Precolumn derivatization of plasma or urine is performed directly in acetonitrile without extraction with p-bromophenacyl bromide and crown ether as catalyst. The p-bromophenacyl ester derivatives are then separated by high-performance liquid chromatography, using an isocratic system of acetonitrile and water containing choline. Effluent was monitored at 254 nm. The limit of detection was 5 micromol/L for betaine and 2 micromol/L for DMG. Analytical recovery was >97% for both analytes. Total and within-day CVs were 2.0-4.4% and 0.9-2.2% for DMG. For betaine, the total and within-day CVs were 1.3-5.3% and 0.4-3.8%, respectively. The method is precise and cost effective and has been used successfully to determine the concentrations of DMG and betaine in human plasma and urine. PMID- 9732981 TI - Evaluation of the tacrolimus II microparticle enzyme immunoassay (MEIA II) in liver and renal transplant recipients. AB - We evaluated the MEIA II with blood samples with added tacrolimus (3.0, 5.0, 11.0, and 22.0 microg/L). The assay had acceptable recoveries (99-103%) and intraday imprecision (<16.0%) across the range of concentrations studied, except for the recoveries at 3.0 microg/L (86.3%) and 5.0 microg/L (80.7%). Comparison of liver (n = 116) and renal (n = 113) patient samples measured by MEIA II against HPLC-tandem mass spectrometry (HPLC-MS/MS) found a mean overestimation of 15.6%. From these comparison data it can be calculated that at values of 5 and 20 microLg/L in liver or renal transplant patient samples, measured by HPLC-MS/MS, MEIA II will have the corresponding range estimates of 3.6-7.9 microg/L and 20.9 25.4 microg/L, respectively. No clinically significant difference in results, in terms of overestimation or correlation, was observed between the two transplant groups studied. The MEIA II is an improvement on the previous MEIA I and is suitable for the therapeutic drug monitoring of tacrolimus where HPLC-MS/MS is unavailable. PMID- 9732982 TI - Serum digoxin in the presence of digibind: determination of digoxin by the Abbott AxSYM and Baxter Stratus II immunoassays by direct analysis without pretreatment of serum samples. AB - We have reevaluated the feasibility of using direct immunochemical methods to track free digoxin in patients receiving Digibind. We report here that results obtained by the Stratus II and AxSYM immunoassays on patients receiving digoxin (without Digibind), digoxin-fortified serum samples supplemented with Digibind, and a digitoxic patient treated with Digibind, show no clinically significant biases. We conclude that useful free digoxin concentrations may be obtained for Digibind-treated patients using either the AxSYM or Stratus immunoassays without subjecting samples to ultrafiltration before analysis. PMID- 9732983 TI - What is hemoglobin A1c? An analysis of glycated hemoglobins by electrospray ionization mass spectrometry. AB - Hemoglobin A1c (HbA1c) is a stable minor Hb variant formed in vivo by posttranslational modification by glucose, originally identified by using cation exchange chromatography, and containing primarily glycated N-terminal beta chains. However, the structure(s) of the quantified species has not been elucidated, and the available methods lack a reference standard. We used electrospray ionization mass spectrometry to determine the extent of glycation of samples separated by boronate affinity and/or cation exchange chromatography. Analyses of clinical samples were consistent with the curvilinear relationship of patient glucose and HbA1c. As glycation increased, the ratio of beta-chain to alpha-chain glycation increased, and the number of glycation sites on the beta chain increased, although these were relatively minor components. We found several glycated species that cochromatographed with HbA1c on cation exchange, including species with both glycated alpha- and beta-chains, nonglycated alpha- and glycated beta-chains, and multiply glycated beta-chains. The combined use of affinity and cation exchange chromatography with structural confirmation by electrospray ionization mass spectrometry was found to be useful in producing samples of sufficient purity for the standardization of glycohemoglobin clinical assays. PMID- 9732984 TI - Multivariate approach to quality control in clinical chemistry. AB - When monitoring analyzer performance in the clinical setting, laboratories are required to test multiple concentrations of control material on a daily basis. Because of the nature of laboratory testing, there is the potential for correlation between the concentrations of control material being monitored. Although traditional clinical quality-control approaches make an underlying assumption of independence with respect to the control concentrations, this will not always be the case. The presence of correlation in some circumstances suggests the use of a new approach for evaluating clinical laboratory monitoring data: the multivariate control chart. Such a chart (the chi2 chart) is evaluated and compared with traditional quality-control approaches used in the laboratory setting. Results indicate that the multivariate approach provides an attractive alternative to many traditional methods of quality assurance when control concentrations are correlated. PMID- 9732986 TI - Measurement of specific immunoglobulin E: intermethod comparison and standardization. AB - Recently introduced "second-generation" techniques for specific IgE measurement have produced some analytical improvement, offering better clinical sensitivity than previous techniques. The aims of our study were to compare the analytical and clinical performances of four second-generation techniques for allergen specific IgE measurement in serum and to ascertain whether the new system for reporting quantitative results contributes to greater clinical agreement between findings using the techniques considered. Allergen-specific IgE was measured using the CAP System, CARLA, ENEA, and AlaSTAT, and the findings were compared. A significant disagreement was found between CAP and ENEA for all allergens and between CAP and CARLA for D1 and G5. However, the clinical discrepancies were reduced by selecting method-specific thresholds using ROC analysis. Second generation techniques enable us to obtain better standardization of results; however, the identification of a specific threshold appears to be a prerequisite for the appropriate clinical interpretation of the test findings. PMID- 9732985 TI - Luminometric single step urea assay using ATP-hydrolyzing urease. AB - An automatic enzyme kinetic luminometric method for determination of small quantities of urea in biological fluids and in microdialysates is presented. The method is based on the ATP-hydrolyzing urease reaction [urea amidohydrolase (ATP hydrolyzing); EC 3.5.1.45], monitored by a luciferin-luciferase ATP reaction. The assay range is 100 pmol to 50 nmol with a detection limit of 5 micromol/L in the sample, compared with detection limits of 0.1 mmol/L in earlier spectrophotometric methods. To reduce the non-urea-dependent ATPase activity (v(blank)) and to increase the urea-dependent activity, 1,2-propanediol was included. Assay conditions were optimized by multivariate analysis. Recoveries of urea added to blood dialysate and plasma were 96-103%. No analytical interference of common metabolites, drugs, or other additives was observed. The total CVs (6 days and six concentrations, 1.2-21.8 mmol/L) were 3.6-8.5%. The results obtained with the present assay were highly correlated for dialysate (r = 0.979) and for plasma (r = 0.978) with those obtained by a spectrophotometric kit method with slopes of 1.02-1.03 and intercepts of 0.08-0.23 mmol/L. PMID- 9732987 TI - False increase in C-reactive protein attributable to heterophilic antibodies in two renal transplant patients treated with rabbit antilymphocyte globulin. AB - Increased serum C-reactive protein (sCRP) is a sensitive marker of renal graft rejection. We describe the cases of two children with uncomplicated renal transplantation who had false-positive sCRP values on analyzers using rabbit anti CRP but values within the reference range with anti-CRP from other animal species. Cross-reaction with heterophilic antibodies was suggested by clinical and biological signs of serum sickness and daily treatment with rabbit antilymphocyte globulin (ALG). The interference depended on the serum concentration of the cross-reactant and was removed by subtotal IgG adsorption to Protein A or Protein G or by immunoadsorption using rabbit ALG or total IgG in non-immune rabbit serum. Anti-rabbit IgG and IgM antibodies were detected in both patients. These are the first reported cases of cross-reaction with heterophilic antibodies in a turbidimetric CRP assay. PMID- 9732988 TI - Fluorescence-based SSCP analysis with automatic allele detection demonstrated for the factor V Leiden mutation. PMID- 9732989 TI - Kinetics of plasma total homocysteine in patients receiving high-dose methotrexate therapy. PMID- 9732990 TI - Improved method to retrieve DNA from dried silver-stained polyacrylamide gels. PMID- 9732991 TI - Prostate-specific antigen expression in neoplastic human myeloid cell lines. PMID- 9732992 TI - Dysfunctional factor VII variant (FVII Tondabayashi) with R79Q: determination of mutated site with monoclonal anti-human factor VII antibody (B101/B1). PMID- 9732993 TI - Serum carcinoembryonic antigen, cancer antigen 125, cancer antigen 15-3, squamous cell carcinoma, and tumor-associated trypsin inhibitor concentrations during healthy pregnancy. PMID- 9732994 TI - Alternative ELISA for sex hormone-binding globulin in plasma. PMID- 9732995 TI - Hemoglobin D [beta 121(GH4)Glu-->Gln] causing falsely low and high HbA1c values in HPLC. PMID- 9732996 TI - Determination of cerebrospinal fluid glucose with the vitros DT60 II dry chemistry system. PMID- 9732997 TI - Comment on the nature and specificity of Bayer Corporation and Chiron Diagnostics hCG immunoassays. PMID- 9732998 TI - Tissue polypeptide antigen as a putative indicator of apoptosis. PMID- 9732999 TI - Miniaturization of analytical systems. AB - Miniaturization has been a long-term trend in clinical diagnostics instrumentation. Now a range of new technologies, including micromachining and molecular self-assembly, are providing the means for further size reduction of analyzers to devices with micro- to nanometer dimensions and submicroliter volumes. Many analytical techniques (e.g., mass spectrometry and electrophoresis) have been successfully implemented on microchips made from silicon, glass, or plastic. The new impetus for miniaturization stems from the perceived benefits of faster, easier, less costly, and more convenient analyses and by the needs of the pharmaceutical industry for microscale, massively parallel drug discovery assays. Perfecting a user-friendly interface between a human and a microchip and determining the realistic lower limit for sample volume are key issues in the future implementation of these devices. Resolution of these issues will be important for the long-term success of microminiature analyzers; in the meantime, the scope, diversity, and rate of progress in the development of these devices promises products in the near future. PMID- 9733000 TI - Ligand assays: from electrophoresis to miniaturized microarrays. AB - The main developments in the "ligand assay" field in which I have been involved are traced. These include the original development of "first generation" competitive assays relying on radiolabeled analyte markers; the development of the first "second generation", noncompetitive (ultrasensitive) methods, which rely on the use of labeled (monoclonal) antibodies and high specific activity nonisotopic labels (leading to the transformation of the immunodiagnostic field in the 1980s); and the development of the first "third generation" miniaturized, chip-based, microarray methods, which permit the simultaneous ultrasensitive measurement of many analytes in the same small sample. The latter--applicable both to immunoassay and to DNA/RNA analysis--are likely to revolutionize the diagnostic and pharmaceutical fields in the next decade. PMID- 9733001 TI - Fixed polarizer ellipsometry for simple and sensitive detection of thin films generated by specific molecular interactions: applications in immunoassays and DNA sequence detection. AB - Biological thin films may form on a surface by specific molecular interactions. The fixed polarizer ellipsometer (FPE) is a sensitive instrument that detects biological thin films either qualitatively or quantitatively. The design is simple and inexpensive. The assays are formatted on an optical surface, and the FPE detection is based on the phase shift of linearly polarized light after reflection through a thin film. We have constructed mathematical models of the FPE response to reflection through single-layer and two-layer films that agree closely with experimental data. Several biological assays have been measured with the FPE to demonstrate the application of this technology to clinical targets, including ultrasensitive immunoassays for hepatitis B surface antigen (0.1 ng/mL) and alpha-fetoprotein (0.01 ng/ mL) and DNA hybridization (0.5 fmol/microL target probe). A clinical study for detection of group A streptococcus from patient throat swabs demonstrated the qualitative application of the FPE to infectious disease targets. The flexibility and sensitivity of the FPE makes this technology suitable for numerous target analytes and applications. PMID- 9733002 TI - Mass-sensing, multianalyte microarray immunoassay with imaging detection. AB - Miniaturization of ligand binding assays may reduce costs by decreasing reagent consumption, but it is less apparent that miniaturized assays can simultaneously exceed the sensitivity of macroscopic techniques by analyte "harvesting" to exploit the total analyte mass available in a sample. Capture reagents (avidin or antibodies) immobilized in 200-microm diameter zones are shown to substantially deplete analyte from a liquid sample during a 1-3-h incubation, and the assays that result sense the total analyte mass in a sample rather than its concentration. Detection of as few as 10(5) molecules of analyte per zone is possible by fluorescence imaging in situ on the solid phase using a near-infrared dye label. Single and multianalyte mass-sensing sandwich array assays of the IgG subclasses show the sensitivity and specificity of ELISA methods but use less than 1/100 the capture antibody required by the 96-well plate format. PMID- 9733003 TI - Cost-efficient use of gas chromatography-mass spectrometry: a "piggyback" method for analysis of gabapentin. PMID- 9733004 TI - Ultrasensitive direct fluorescent immunoassay for thyroid stimulating hormone. PMID- 9733005 TI - Charge coupled device optics system for simultaneous measurement of multiple reactions in a microplate. PMID- 9733006 TI - Determination of the viability of Escherichia coli at the single organism level by electrorotation. PMID- 9733007 TI - A miniaturized, self-contained, single-use, disposable assay device for the quantitative determination of the bone resorption marker, NTx, in urine. PMID- 9733008 TI - Simultaneous detection of multiple analytes using Copalis technology: a reduction to practice. PMID- 9733009 TI - A rapid, sensitive, multiplexed assay for detection of viral nucleic acids using the FlowMetrix system. PMID- 9733010 TI - Clinical evaluation of a new, miniaturized biosensor for self-monitoring of blood glucose. PMID- 9733011 TI - Multiplexed analysis of human cytokines by use of the FlowMetrix system. PMID- 9733012 TI - New approaches in the binding of DNA for clinical applications. PMID- 9733013 TI - Transference of reference intervals in the validation of automated chemiluminescent immunoassays on a new platform. PMID- 9733014 TI - Fully automated enzyme immunoassay system for the determination of activator specific histamine release from basophils in whole blood. PMID- 9733015 TI - Chemiluminescent detection of DNA in low- and medium-density arrays. PMID- 9733016 TI - Oxidative stress in chronic renal failure. AB - Cardiovascular disease is the major cause of morbidity and mortality in chronic renal failure. The aim of this review is to summarise current evidence suggesting that there is increased free radical production, antioxidant depletion and changes in lipoprotein composition in renal failure which will lead to oxidation of LDL and hence to accelerated development of atherosclerosis. PMID- 9733017 TI - Mechanism of reaction of nitrogen dioxide radical with hydroxycinnamic acid derivatives: a pulse radiolysis study. AB - Nitrogen dioxide radical (NO2.) is known as a toxic agent produced in the metabolism of nitrates and nitrites. By the use of the pulse radiolysis technique, the mechanism of the reaction of NO2. radical with hydroxycinnamic acid derivatives (HCA) was studied and the rate constants have been measured. The rate constants were found to be 7.4 x 10(8), 7.2 x 10(8), 8.6 x 10(8) dm3 mol(-1) s(-1) for ferulic acid, sinapic acid and caffeic acid, respectively. The reactions produce the corresponding phenoxyl radical. PMID- 9733018 TI - Oxidative stress in subjects affected by celiac disease. AB - In order to study the role of oxidative stress in celiac disease, protein carbonyl groups, thiobarbituric acid-reactive substance and pentosidine were evaluated in the plasma of nine patients with asymptomatic celiac disease and in a control group (n = 25). Plasma alpha-tocopherol, retinol and lipids were determined in the same samples. The levels of markers of oxidative stress derived from both protein (carbonyl groups) and lipids (thiobarbituric acid-reactive substances) were significantly higher in celiac disease patients, whereas lipoproteins and alpha-tocopherol were significantly lower. These data indicate that in celiac disease, even when asymptomatic, a redox imbalance persists; this is probably caused by an absorption deficiency, even if slight. Dietary supplementation with antioxidant molecules may offer some benefit and deserves further investigation. PMID- 9733019 TI - Different DNA damaging species as a result of oxidation of n-butyraldehyde and iso-butyraldehyde by Cu(II). AB - The isomers n- and iso-butyraldehyde (BuA) in combination with Cu(II) induced single and double strand breaks in PM2 DNA, whereas the aldehydes, or Cu(II) alone had only negligible effect. The DNA damage was the result of radical oxidations of the aldehydes under formation of Cu(I). Cu(I) formation was independent of molecular oxygen. Extensive DNA degradation was only observed in the presence of molecular oxygen. Characterization of DNA damage pointed to different ultimate DNA damaging species. While catalase and neocuproine inhibited strand break formation induced by iso-BuA/Cu(II) to a high degree, these inhibitors were less effective in the n-BuA/Cu(II) reaction. On the other hand, sodium azide showed a high strand break inhibition in the n-BuA/Cu(II) reaction, but low inhibition in the iso-BuA/Cu(II) reaction. 2-Deoxyguanosine was hydroxylated in the 8-position by iso-BuA/Cu(II) but little reaction occurred with n-BuA/Cu(II). Chemiluminescence was detected during both BuA/Cu(II) reactions, whereby the intensity of the luminescence signal was 3.5-fold higher for n-BuA/Cu(II) than for iso-BuA/Cu(II). We suppose that the copper(II)-driven oxidation of n- and iso-BuA proceeds via different pathways with different DNA damaging consequences. Whereas the oxidation of iso-BuA mainly results in damage by .OH-radicals, the oxidation of n-BuA may lead to a radical reaction chain whereby excited states are involved and the resulting DNA-damaging species are not .OH-radicals. PMID- 9733021 TI - Oxidation of lipids in low density lipoprotein particles. AB - This study was undertaken to understand further the mechanisms and dynamics of the oxidation of lipids in low density lipoprotein (LDL) particles, aiming specifically at elucidating the material balance between oxygen uptake and products found and also the relative susceptibilities to oxidation of cholesteryl ester in the core and phosphatidylcholine in the outer monolayer in the LDL particles. It was found that considerable amount of oxygen uptake could not be accounted for by conjugated diene or total peroxides. Total peroxide was measured from the phosphine oxide formed from triphenylphosphine or diphenyl pyrenylphosphine by reduction of peroxides. Cholesteryl ester hydroperoxides and phosphatidylcholine hydroperoxides were the major peroxides formed in LDL oxidation, but they accounted for about 60% of total peroxide. Cholesterol was also oxidized, but its oxidation was significant only at the later stages of the reaction. It was also found that the oxidizability of cholesteryl ester relative to phosphatidylcholine was larger within the LDL particle than in homogeneous solution and this was interpreted in the context of the physical properties of LDL particle. PMID- 9733020 TI - Liver microsomal parameters related to oxidative stress and antioxidant systems in hyperthyroid rats subjected to acute lindane treatment. AB - Liver microsomal functions related to xenobiotic biotransformation and free radical production were studied in control rats and in animals subjected to L 3,3',5-triiodothyronine (T3) and/or lindane administration as possible mechanisms contributing to oxidative stress, in relation to the activity of enzymes (superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx), and glucose 6-phosphate dehydrogenase (G-6PDH)) and content of lipid-soluble vitamins (alpha tocopherol, beta-carotene, and lycopene) affording antioxidant protection. Lindane treatment in euthyroid rats at a dosage of 20mg/kg did not modify the content of liver microsomal cytochromes P450 and b5, the activity of NADPH cytochrome P450 reductase and NADH-cytochrome b5 reductase, and the production of superoxide radical (O2.-), as well as antioxidant systems, except for the reduction in lycopene levels. Hyperthyroidism elicited a calorigenic response and increased specific and molecular activities of NADPH-cytochrome P450 reductase, O2.- generation, and G-6PDH activity, concomitantly with diminution in liver SOD and catalase activities and in alpha-tocopherol, beta-carotene, and lycopene levels. The administration of lindane to hyperthyroid animals led to a further increase in the molecular activity of NADPH-cytochrome P450 reductase and in the O2.- production/SOD activity ratio, and decrease of hepatic alpha-tocopherol content, in a magnitude exceeding the sum of effects elicited by the separate treatments, as previously reported for reduced glutathione depletion. Collectively, these data support the contention that the increased susceptibility of the liver to the toxic effects of acute lindane treatment in hyperthyroid state is conditioned by potentiation of the hepatic oxidative stress status. PMID- 9733023 TI - Radiation-induced effects on cefotaxime: ESR study. AB - As an alternative to heat and gas exposure sterilization, ionizing radiation is gaining interest as sterilization process for medicinal products. Detection and dosimetry of pharmaceuticals radiosterilization is a growing concern to numerous government regulatory agencies worldwide. In this context, it is necessary to find methods distinguishing between irradiated and nonirradiated pharmaceuticals. In the absence of suitable detection methods, our attention was focused on electron spin resonance (ESR) spectrometry. A third generation cephalosporin, cefotaxime, was chosen as model; this antibiotic is a potential candidate for radiation treatment due to its thermosensitivity. While the ESR spectra of a nonirradiated sample presents no signal, a signal, dependent of the irradiation dose, is found in irradiated samples. The number of free radicals was estimated by comparing the second integral from radiosterilized samples and a diphenyl picrylhydrazyl reference. Estimation of the number of free radicals gives 1.9 x 10(20) radicals mol(-1) at 20 kGy. From this result, the G-value (number of radicals (100eV)(-1)) could be estimated to 0.3. Aside from qualitative detection, ESR spectrometry can be used for dose estimation. When quadratic, exponential or bi-exponential functions are applied to the variation of peak to peak amplitude vs. dose, these functions correlate well with the data. However, it is important to notice that linear function correlates well with the data for doses lower than 20 kGy. Since the radiation dose selected must be always based upon the bioburden of the products and the degree of sterility required (EN 552 and ISO 11137) 25kGy could no longer be accepted as a "routine dose" for sterilizing a pharmaceutical. Doses from 6kGy (ISO 11137) could be investigated and linear regression would appear to be the least expensive route to follow. The free radicals concentration appeared to not decrease during the 57 days of storage; the number generated during the irradiation allows the detection of radiosterilized cefotaxime up to two years after irradiation. PMID- 9733022 TI - Antioxidant activity of 5-aminosalicylic acid against peroxidation of phosphatidylcholine liposomes in the presence of alpha-tocopherol: a synergistic interaction? AB - Oxidative damage has been implicated in the pathogenesis of inflammatory bowel diseases. 5-Aminosalicylic acid (5-ASA), the anti-inflammatory drug commonly used in the treatment of this condition, has been shown to possess antioxidant properties considered to be of particular importance in the pathologic context of these diseases. However, its action mechanisms are far from being completely elucidated, especially regarding its antioxidant properties in the presence of endogenous antioxidants such as alpha-tocopherol (alpha-T), the major defence system of biomembranes against lipid peroxidation. In this study we investigated the scavenging activity of 5-ASA toward peroxyl radicals generated at different sites of soybean PC liposomes, used as model membranes, either alone or in combination with alpha-T. 5-ASA, separately, shows strong scavenging activity toward peroxyl radicals generated in the aqueous phase by thermal decomposition of 2,2'-azobis(2-amidinopropane hydrochloride) (AAPH), inducing a clear concentration-dependent inhibition period, either of oxygen consumption or of conjugated diene hydroperoxides production. HPLC analysis indicates that 5-ASA is consumed, at a constant rate, throughout the reaction, and when the inhibition period is over, the oxidation rate is resumed. On the other hand, apart from a slight decrease in the rate of oxidation, 5-ASA is unable to suppress efficiently lipid peroxidation, when the reaction starts inside the lipid membranes, by thermal decomposition of 2,2'-azobis(2,4-dimethylvaleronitrile) (AMVN). When 5 ASA is combined with alpha-T, and the oxidation starts in the aqueous phase, an additive inhibitory effect occurs between both compounds. 5-ASA protects efficiently alpha-T against initial attack from AAPH-peroxyl radicals, delaying its consumption. On the other hand, if the reaction starts inside the lipid bilayer, 5-ASA prolongs significantly the inhibitory period produced by alpha-T on the initial rate of oxidation, as measured by oxygen consumption and conjugated diene hydroperoxides. This inhibitory effect points to a synergistic interaction between 5-ASA and alpha-T, since 5-ASA, by itself, is unable to suppress the oxidation reaction. Therefore, 5-ASA reveals an important cooperative effect with alpha-T, either affording an efficient protection to this antioxidant compound, when free radicals are generated in the aqueous site, or potentiating its activity when oxidation is initiated inside the lipid bilayer. Taking into account that the ascorbic acid content decreases significantly in the inflamed mucosa of patients with inflammatory bowel diseases, our data are, certainly, a very important contribution to the knowledge of the anti inflammatory action of 5-ASA. PMID- 9733024 TI - Monitoring of low density lipoprotein oxidation by low-level chemiluminescence. AB - A method for monitoring low-density lipoprotein (LDL) oxidation by low-level chemiluminescence (LL-CL) is described in this study. The kinetic indices obtained with this procedure, in particular lag-time and K value (related to prooxidant activity of Cu2+ bound to LDL) are compared with those of the established UV-absorbing conjugated diene assay. The correlation of lag-time values obtained by LL-CL and conjugated diene assay was very high both in the case of Cu2+- and peroxyl-radical-mediated oxidation (r = 0.99). By using the transient free radical scavenging activity of butylated hydroxytoluene, a calibration of LL-CL for lipid peroxyl radical and termination rate was obtained. The spectral analysis of LL-CL from oxidizing LDL shows a maximum peak between 420 and 500 nm, corresponding to the emission of triplet carbonyl compounds. LL CL allows continuous and direct monitoring of LDL oxidation as extraction and derivatization of lipid peroxidation products are not required. Moreover, some limitations of UV spectroscopy such as by absorbing compounds need not be considered. Therefore, the present procedure represents a simple and convenient tool for continuous monitoring of LDL oxidation which may be applied to mechanistic and clinical studies. PMID- 9733025 TI - Effect of combined coenzyme Q10 and d-alpha-tocopheryl acetate supplementation on exercise-induced lipid peroxidation and muscular damage: a placebo-controlled double-blind study in marathon runners. AB - To test the effects of combined coenzyme Q10 (Q10) and d-alpha-tocopheryl acetate supplementation on exercise-induced oxidative stress and muscular damage we conducted a double-blind study in 37 moderately trained male marathon runners. These were randomly allocated to receive either an antioxidant cocktail: 90 mg of Q10 and 13.5 mg of d-alpha-tocopheryl acetate daily (18 men) or placebo (19 men) for three weeks before a marathon (42km) run. Just before the run, plasma Q10 was 282% (p < 0.0001) and plasma vitamin E 16% (p < 0.007) higher in the supplemented group, than in the placebo group. Also the proportion of plasma ubiquinol of total Q10, an indication of plasma redox status in vivo, was significantly higher in the supplemented group. Furthermore, the susceptibility of the VLDL + LDL fraction, to copper-induced oxidation, was significantly reduced in the supplemented group, compared to the placebo group. The exercise increased lipid peroxidation significantly in both study groups, as assessed by the elevated proportion LDL of LDL and the increased susceptibility of lipoproteins to copper induced oxidation. However, the supplementation had no effect on lipid peroxidation or on the muscular damage (increase in serum creatine kinase activity or in plasma lactate levels) induced by exhaustive exercise. Plasma ascorbate, Q10, whole blood glutathione and serum uric acid concentrations increased during the exercise, elevating significantly the TRAP value of plasma by 10.3% and the proportion of plasma ubiquinol of total Q10 by 4.9%. These results suggest that even though exercise increases plasma lipid peroxidation, it also elevates the antioxidative capacity of plasma, as assessed by the increased plasma TRAP and the proportion of Q10H2 of total Q10. However, prior supplementation with small doses of Q10 and d-alpha-tocopheryl acetate neither attenuates the oxidation of lipoproteins nor muscular damage induced by exhaustive exercise such as encountered in a marathon run. PMID- 9733026 TI - Medical complications of achondroplasia: a multicentre patient review. AB - Achondroplasia is the most prevalent chondrodysplasia and numerous authors have documented the varied social and medical complications that may compromise a full and productive life. Complications include cervicomedullary compression, spinal stenosis, restrictive and obstructive lung disease, otitis media, and tibial bowing, among others. These known complications have led to recommendations for the anticipatory management of such patients. There are relatively few data on the actual rates and timing of these problems. This paper reports data on the rates and age of occurrence of several of these complications based on a review of recorded chart information of 193 patients ascertained from several well established genetic centres with a known interest in the chondrodysplasias. The length of follow up varied and the rates of occurrence at specific age intervals were used to estimate the cumulative percentage affected for each complication. The report includes information on otitis media, ventilation tubes, hearing loss, tonsillectomy, speech problems, tibial bowing and osteotomy, ventricular shunting, apnoea, cervicomedullary decompression, and neurological signs attributable to spinal stenosis. PMID- 9733027 TI - Sperm DNA analysis in a Friedreich ataxia premutation carrier suggests both meiotic and mitotic expansion in the FRDA gene. AB - Friedreich ataxia is usually caused by an expansion of a GAA trinucleotide repeat in intron 1 of the FRDA gene. Occasionally, a fully expanded allele has been found to arise from a premutation of 100 or less triplet repeats. We have examined the sperm DNA of a premutation carrier. This man's leucocyte DNA showed one normal allele and one allele of approximately 100 repeats. His sperm showed an expanded allele in a tight range centering on a size of approximately 320 trinucleotide repeats. His affected son has repeat sizes of 1040 and 540. These data suggest that expansion occurs in two stages, the first during meiosis followed by a second mitotic expansion. We also show that in all informative carrier father to affected child transmissions, with the notable exception of the premutation carrier, the expansion size decreases. PMID- 9733028 TI - Ovine neuronal ceroid lipofuscinosis: a large animal model syntenic with the human neuronal ceroid lipofuscinosis variant CLN6. AB - The neuronal ceroid lipofuscinoses (NCLs) are a group of inherited degenerative neurological diseases affecting children. A number of non-allelic variants have been identified within the human population and the genes for some of these have recently been identified. The underlying mechanism for the neuropathology remains an enigma; however, pioneering studies with the naturally occurring ovine model (OCL) have led to the proposal that these diseases represent lesions in specific hydrophobic protein degradation pathways. In this study, we show linkage between OCL and microsatellite markers on OAR 7q13-15. Using interspecies chromosome painting we establish that OAR 7q13-15 is syntenic with human chromosome 15q21 23, the region which was recently defined as the location of a newly identified late infantile variant (CLN6). We propose that our ovine model represents a mutation in the gene orthologous to that mutated in the human late infantile variant CLN6. The ovine linkage flock, consisting of 56 families, represents a powerful resource for positional cloning of this NCL gene. The availability of such a large animal model will have important implications for experimentation in downstream corrective therapies. PMID- 9733029 TI - Del(18p) shown to be a cryptic translocation using a multiprobe FISH assay for subtelomeric chromosome rearrangements. AB - We have previously described a fluorescence in situ hybridisation (FISH) assay for the simultaneous analysis of all human subtelomeric regions using a single microscope slide. Here we report the use of this multiprobe FISH assay in the study of a patient whose karyotype was reported by G banding analysis as 46,XX,del(18)(p11.2). Although the proband had some features suggestive of a chromosomal abnormality, relatively few of the specific features of del(18p) were present. She was a 37 year old female with mild distal spinal muscular atrophy (SMA), arthritis of the hands, an abnormal chest shape (pectus excavatum), and an unusual skin condition (keratosis pilaris). Reverse chromosome painting with degenerate oligonucleotide primer-polymerase chain reaction (DOP-PCR) amplified del(18p) chromosomes as a probe confirmed the abnormality as del(18p), with no evidence of any other chromosome involvement. Subsequently, the multiprobe FISH assay confirmed deletion of 18p subtelomeric sequence. However, the assay also showed that sequences corresponding to the 2p subtelomeric probe were present on the tip of the shortened 18p. The patient is therefore monosomic for 18p11.2-pter and trisomic for 2p25-pter, and the revised karyotype is 46,XX,der(18)t(2;18)(p25; p11.2). We believe that a proportion of all cases reported as telomeric deletions may be cryptic translocations involving other chromosome subtelomeric regions. Further studies such as this are necessary to define accurately the clinical characteristics associated with pure monosomy in chromosomal deletion syndromes. PMID- 9733030 TI - Molecular, cytogenetic, and clinical characterisation of six XX males including one prenatal diagnosis. AB - Cytogenetic analysis, fluorescent in situ hybridisation (FISH), and molecular amplification have been used to characterise the transfer of Yp fragments to Xp22.3 in six XX males. PCR amplification of the genes SRY, RPS4Y, ZFY, AMELY, KALY, and DAZ and of several other markers along the Y chromosome short and long arms indicated the presence of two different breakpoints in the Y fragment. However, the clinical features were very similar in five of the cases, showing a male phenotype with small testes, testicular atrophy, and azoospermia. All these patients have normal intelligence and a stature within the normal male range. In the remaining case, the diagnosis was made prenatally in a fetus with male genitalia detected by ultrasound and a 46,XX karyotype in amniocytes and fetal blood. Molecular analysis of fetal DNA showed the presence of the SRY gene. FISH techniques also showed Y chromosomal DNA on Xp22.3 in metaphases of placental cells. To our knowledge, this is the second molecular prenatal diagnosis reported of an XX male. PMID- 9733031 TI - Family history of breast cancer: what do women understand and recall about their genetic risk? AB - The current study has two aims: (1) to look at people's recall of risk information after genetic counselling and (2) to determine the impact of receiving an audiotape of the genetic consultation on level of recall, cancer related worry, and women's uptake of risk management methods. Using a prospective randomised controlled design, subjects receiving an audiotape were compared with a standard consultation group. Participants were drawn from attenders at the genetic clinics of two London hospitals and included 115 women with a family history of breast cancer. Assessment of perceived genetic risk, mental health, cancer worry, and health behaviour was made before counselling at the clinic (baseline) and by postal follow up. Usefulness of audiotapes and satisfaction with the clinical service was assessed by study specific measures. The data indicate that cancer worry is reduced by provision of an audiotape of the genetic consultation. Recall of the genetic risk figure, however, is not affected by provision of an audiotape and neither is it related to women's overall perception of being more or less at risk of breast cancer than the average woman. Forty-one percent of women accurately recalled their personal risk of breast cancer at one month follow up; however, 25% overestimated, 11% underestimated, and 23% could not remember or did not know their breast cancer risk. Recall of the risk figure is more accurate when the clinical geneticist has given this to the woman as an odds ratio rather than in other formats. Subsequent health behaviour is unaffected by whether women have an audiotape record of their genetic consultation. Results suggest that having a precise risk figure may be less important than women taking away from the consultation an impression that something can be offered to help them manage that risk. Provision of an audiotape of the consultation is of limited usefulness. The need for psychological care to be better integrated into genetic counselling at cancer family clinics was highlighted by the study. The results are discussed in terms of future service development. PMID- 9733032 TI - Men in breast cancer families: a preliminary qualitative study of awareness and experience. AB - In inherited forms of breast cancer, attention in clinical genetics services has focused on women because they are most at risk of developing cancer. Men at risk of transmitting a predisposing gene mutation are less likely to have a genetic test than the women in these families. This preliminary study investigates the perspective of the brothers of women with familial breast cancer and is based on qualitative analysis of 22 semistructured interviews using an attenuated form of Grounded Theory. There is an awareness among these men (without having had genetic counselling) that the breast cancer in their families is inherited. Some of them harbour fear of developing cancer themselves and many are concerned that their daughters might develop breast cancer. Some appeared to use avoidance as a coping strategy. The men were very often excluded from family conversations about breast cancer. Implications for the provision of genetic counselling for these families are discussed. PMID- 9733034 TI - Four sibs with dislocated elbows, bowed tibiae, scoliosis, deafness, cataract, microcephaly, and mental retardation: a new MCA/MR syndrome. AB - We report four sibs with an MCA/MR syndrome whose parents were first cousins. The sibs had mental retardation, microcephaly, hearing problems, cataract, and multiple osseous malformations, such as dislocated elbows, bowed tibiae, and scoliosis. Review of published reports and the use of the London Dysmorphology Database suggest that this family presents a new syndrome. PMID- 9733033 TI - Predicting adaptation to presymptomatic DNA testing for late onset disorders: who will experience distress? Rotterdam Leiden Genetics Workgroup. AB - The first comparative study on predicting post-test distress (conceptualised by intrusion and avoidance, measured with the Impact of Event Scale) after presymptomatic genetic testing for Huntington's disease (HD, n=25), cancer syndromes (familial adenomatous polyposis (FAP, n=23)), and hereditary breast and ovarian cancer (HBOC, n=10) is reported. The variables with the highest predictive potential of post-test distress are presented. Participants who were depressed before the test were more distressed after testing, but we found that those who were anxious before the test were less distressed, that is, had less intrusive thoughts post-test. Other factors associated with a higher level of post-test intrusion were gender (being a woman), having children, and pre-test intrusion. Religion and being at risk for HBOC were associated with less post test intrusion. Participants who showed avoidance behaviour before the test and those who had many people available for support showed more avoidance behaviour post-test. The test result did not additionally contribute to post-test distress. The prima facie simple notion that the test result, as such, determines the distress experienced seems to be a misrepresentation of the complex reality. PMID- 9733035 TI - Spastic paraplegia, optic atrophy, microcephaly with normal intelligence, and XY sex reversal: a new autosomal recessive syndrome? AB - Two female sibs of first cousin Iranian parents were found to have the syndrome of spastic paraplegia, optic atrophy with poor vision, microcephaly, and normal cognitive development. Karyotype analysis showed a normal female constitution in one and a male constitution (46,XY) in the other. The XY female showed normal female external genitalia, normal uterus and tubes, and streak gonads. SRY gene sequencing was normal. We conclude that the present family probably represents a new autosomal recessive trait of pleiotropic effects including XY sex reversal and adds further evidence for the heterogeneity of spastic paraplegia syndromes as well as sex reversal syndromes. PMID- 9733036 TI - Familial craniosynostosis, anal anomalies, and porokeratosis: CAP syndrome. AB - We report on the occurrence of coronal craniosynostosis, anal anomalies, and porokeratosis in two male sibs. A third male sib was phenotypically normal as were the parents. The occurrence of these three clinical features has, to our knowledge, not been reported before. Cutaneous or anal anomalies or both have been reported in a number of syndromes associated with craniosynostosis, including Crouzon, Pfeiffer, Apert, and Beare-Stevenson syndromes. These syndromes are associated with mutations in the fibroblast growth factor receptor genes FGFR1, FGFR2, and FGFR3. They are inherited in an autosomal dominant fashion. In contrast, the cases we report do not carry any of the common FGFR mutations and the pedigree suggests autosomal or X linked recessive inheritance. PMID- 9733037 TI - Baller-Gerold syndrome associated with congenital portal venous malformation. AB - We report a 4 year old boy in whom the clinical features of craniosynostosis and bilateral absent radii led to a diagnosis of Baller-Gerold syndrome. Additional congenital abnormalities included midface hypoplasia, atrial and ventricular septal defects, right hydronephrosis, partial sacral agenesis, and anterior ectopic anus. Evidence of portal venous hypertension was present from 8 months and a congenital portal venous malformation was discovered at 2 years. This is the first reported case of Baller-Gerold syndrome associated with a congenital portal venous malformation. We discuss the diagnostic confusion between this syndrome and other overlapping malformation syndromes and propose optimal evaluation strategies aimed at clarifying the nosology of these syndromes. PMID- 9733038 TI - Fine localisation of the gene for central areolar choroidal dystrophy on chromosome 17p. AB - Central areolar choroidal dystrophy (CACD) is a retinal disease which causes progressive profound loss of vision in patients during middle age. The disease is inherited as an autosomal dominant trait and shows genetic heterogeneity. Mutations in the peripherin-RDS gene on chromosome 6 have been reported in affected members of families transmitting the disease. A new locus at chromosome 17p13 was identified recently by a genome wide linkage search in members of a large Northern Irish family. We now report the refinement of the critical region for this gene to an interval of approximately 5 cM flanked by polymorphic markers D17S1810 and CHLC GATA7B03. PMID- 9733039 TI - Further refinement of the Usher 2A locus at 1q41. AB - Usher syndrome (USH) is characterised by congenital sensorineural hearing loss and progressive pigmentary retinopathy. All three subtypes (USH1, USH2, and USH3) are inherited as recessive traits. People with Usher type 2 (USH2) have normal vestibular responses and moderate to severe hearing loss. These syndromes have been found to be genetically heterogeneous, with a single locus for USH2 at 1q41 (USH2A), six loci for USH1, and one for USH3. Some USH2 families have been excluded from the 1q41 locus suggesting that a second, as yet unidentified, locus (USH2B) must exist. Linkage studies suggest that around 90% of USH2 families are USH2A. Four USH2 families were analysed for linkage to markers flanking the USH2A locus. In one of these families a recombination event was observed in an affected subject which excludes the USH2A gene from proximal to the marker AFM143XF10 and defines this as the new centromeric flanking marker for the USH2A locus. A further recombination event in another patient from this family confirmed AFM144XF2 as the telomeric flanking marker. The interval between these polymorphic markers is estimated to be 400 kb. This region is completely contained in each of three YACs from the CEPH library: 867g9, 919h3, and 848b9. This refinement more than halves the critical genetic interval and will greatly facilitate positional cloning of the USH2A gene. PMID- 9733040 TI - Recurrence of the D409H mutation in Spanish Gaucher disease patients: description of a new homozygous patient and haplotype analysis. AB - Gaucher disease results, in most patients, from mutations in the gene encoding glucocerebrosidase. Mutation D409H is the third most frequent in Spanish patients, accounting for 5.7% of all mutated alleles. This allele is associated mainly with the neurological forms of the disease. Recently, homozygosity for the D409H mutation has been associated with a particular phenotype, including specific cardiovascular symptoms. Here we report a second Spanish patient bearing the D409H/D409H genotype with a very early manifestation of the disease. The patient started enzyme replacement therapy at 3 months of age. A common origin for the Spanish D409H alleles was ruled out by haplotype analysis using an internal polymorphism of the glucocerebrosidase gene and two external microsatellite markers. PMID- 9733041 TI - Identical de novo mutation at the D4F104S1 locus in monozygotic male twins affected by facioscapulohumeral muscular dystrophy (FSHD) with different clinical expression. AB - Facioscapulohumeral muscular dystrophy (FSHD) is a progressive hereditary neuromuscular disorder, transmitted in an autosomal dominant fashion. Its clinical expression is highly variable, ranging from almost asymptomatic subjects to wheelchair dependent patients. The molecular defect has been linked to chromosome 4q35 markers and has been related to deletions of tandemly repeated sequences located in the subtelomeric region detected by probe p13E-11 (D4F104S1). We describe a pair of monozygotic male twins affected by FSHD, carrying an identical de novo p13E-11 EcoRI fragment of paternal origin and showing great variability in the clinical expression of the disease, one being almost asymptomatic and the other severely affected. Their medical history was the same, with the exception of an anti-rabies vaccination performed at the age of 5 in the more severely affected twin. We hypothesise that the vaccination might have triggered an inflammatory immune reaction contributing to the more severe phenotype. PMID- 9733042 TI - Paternally inherited deletion of CSH1 in a patient with Silver-Russell syndrome. AB - In a continuing study on the aetiology of Silver-Russell syndrome (SRS), we detected a patient with a heterozygous deletion in the growth hormone gene cluster (17q22-q24). The deletion of the chorionic somatomammotrophin hormone 1 (CSH1) gene was inherited from the patient's father. The patient shows typical symptoms of SRS. Though deletions of CSH1 have been reported without any phenotypic consequences, the heterozygous deletion might be involved in the aetiology of SRS in the case presented here. Apart from other observations in SRS, like maternal uniparental disomy 7, changes in the genomic region 17q22-qter might be responsible for the expression of this syndrome for at least some of the patients, leading to the heterogeneity of SRS. PMID- 9733043 TI - A new dominant retinitis pigmentosa family mapping to the RP18 locus on chromosome 1q11-21. PMID- 9733044 TI - Frequency of inherited deletions of 22q11. PMID- 9733045 TI - The annual incidence of DiGeorge/velocardiofacial syndrome. PMID- 9733046 TI - Sharing of PPT mutations between distinct clinical forms of neuronal ceroid lipofuscinoses in patients from Scotland. PMID- 9733047 TI - PTEN and prostate cancer. PMID- 9733048 TI - Instability in the normal CTG repeat range at the myotonic dystrophy locus. PMID- 9733049 TI - Outcome of combination chemotherapy in extensive stage small-cell lung cancer: any treatment related progress? AB - During the past two decades many different treatment regimens of combination chemotherapy have been applied in extensive stage small-cell lung cancer (SCLC). This study was carried out to identify whether these modifications have resulted in an improved overall survival for extensive stage during the past two decades. In total, 1111 patients with extensive stage SCLC were included in six consecutive randomised trials in our setting from 1973 until 1992. Of these, 526 patients treated in the early period (1973-1981) were compared with 585 patients treated in the late period (1981-1992) with respect to pretreatment prognostic factors, staging, treatment and outcome. No change in the distribution of prognostic factors was detected and the frequency of patients with extensive stage was equal in the two periods, and no difference in overall response rates and survival was observed (P = 0.49). Median survival in the two periods was 208 days and 215 days, respectively. No stage migration or treatment-related improved outcome was observed in extensive disease. We suggest restricting aggressive treatment to patients with favorable prognosis and long-term survival as a realistic aim. PMID- 9733050 TI - Criteria of functional and oncological operability in surgery for lung cancer: a multicenter study. The Bronchogenic Carcinoma Cooperative Group of the Spanish Society of Pneumology and Thoracic Surgery (GCCB-S). AB - This study was undertaken to determine how much agreement was prevalent in the criteria of functional operability (a patient's capacity for tolerating surgery) and oncological operability (the preoperative assessment of the potential for excising all tumoral tissue with a prognostically favorable result) used in the hospitals participating in the Bronchogenic Carcinoma Cooperative Group of the Spanish Society of Pneumology and Thoracic Surgery (GCCB-S). The GCCB-S includes 17 hospitals in which all consecutive cases of lung cancer in which thoracotomy is performed have been registered since 1993. A survey was sent to all participating centers requesting a semiquantitative evaluation of the level of agreement between the functional and oncological operability criteria used in the hospital and the functional and oncological operability criteria established by consensus among the members of the Coordinating Group of the GCCB-S. The consensus criteria were established mainly by evaluating evidence from the literature. Fourteen hospitals completed the survey: in 13 hospitals (93%) the consensus functional operability criteria were used 'just about always' (in more than 90% of cases) and in one hospital 'almost always' (between 80 and 90% of cases). In 14 hospitals the consensus oncological operability criteria were used 'just about always' (more than 90% of cases), although in five centers (36%) other tests, mainly abdominal imaging, were also used systematically. In conclusion, this GCCB-S study detected a broad agreement among the participating hospitals as to the minimal criteria for preoperative evaluation of the operability of patients with lung cancer. PMID- 9733051 TI - Radiation therapy alone in the treatment of tumours of the trachea. AB - A retrospective analysis of 23 patients with tracheal malignancy treated with a radiation therapy alone is reported. All patients were irradiated at Bydgoszcz Cancer Center during the period 1990-1996. To overcome serious damage to normal tissues, a dose escalation combination of external beam irradiation and brachytherapy was used in most cases. Squamous cell carcinoma was the most common type and was seen in 13 cases. Adenoid cystic carcinoma occurred in seven, adenocarcinoma in two and carcinoid in one patient. Eight patients were treated with definitive and 15 with palliative intent. Local control was attained in 8 of 23 patients and was more frequent for patients from curative group treated with doses greater than 60 Gy. The mean survival for all patients was 9.5 months, and 26 and 7.2 months for definitive and palliative group, respectively. Survival was strongly correlated to histologic type and response to radiotherapy. PMID- 9733052 TI - Sera from patients with malignant mesothelioma can contain autoantibodies. AB - Malignant mesothelioma (MM) is resistant to all conventional forms of therapy though there is considerable evidence from clinical trials and animal models of the disease that an immune response can be elicited to the tumour. In order to define those target antigens expressed by MM cells which might provide a focus for an effective immune response we tested patients' sera for the presence of MM autoantibodies by Western blot analysis. Eight of 29 (28%) patients with MM had serum antibodies of the IgG class in high titre and each antiserum recognised different protein antigens. In those individuals where sequential samples were available, the antibody titre increased with the progression of the disease though the number of target antigens remained constant. Sera from the eight patients were studied further: six of the antigen complexes were expressed at least partially in the nucleus; two showed some specificity for the tumour in that they discriminated antigens that were highly expressed in all human MM cell lines, but were not expressed in a human SV40 transformed mesothelial line; four of the antisera recognised a homologue in mouse tissue and each of these had a different pattern of expression. Collectively, these antisera define a subset of nuclear autoantigens that are over-expressed in dividing cells. PMID- 9733053 TI - HLA class I and class II expression of pulmonary adenocarcinoma cells and the influence of interferon gamma. AB - BACKGROUND: The clinico-biological significance of HLA (both class I antigen and class II one) expressed on tumor cells still remains controversial. METHODS: Tumor cells were freshly separated from 33 surgical specimens of pulmonary adenocarcinoma. The tumor cells were incubated for 24 h in the presence or absence of IFN-gamma (130 International Units/ml). After incubation, the cells were cytocentrifuged onto glass slides and immunostained with either an anti-HLA class I (A, B, C) monoclonal antibody or anti-HLA class II (DR) one. RESULTS: In 22 of 33 cases (66.7%), the HLA class I were individually expressed by more than 60% of tumor cells while so were the HLA class II in 15 (45.4%). No significant correlation was observed between the HLA class I expression and the HLA class II one. The proportion of HLA class I-positive tumor cells correlated with neither the grade of histological differentiation nor the stage of disease. In contrast, the proportion of HLA class II-positive tumor cells correlated with both the grade of histological differentiation and the stage. In most cases, IFN-gamma was found to increase the proportion of class II-positive tumor cells as well as that of class I-positive cells. CONCLUSIONS: The above findings thus suggested that the HLA class II expression might therefore represent a manifestation of cellular differentiation and that IFN-gamma may, as a result, have the potential to differentiate cancer cells. PMID- 9733054 TI - Phase-II randomized study of pre-operative IL-2 administration in operable NSCLC. AB - Lymphocytopenia is a prognostic factor for shorter survival in advanced lung cancer and it is likely related to an interleukin-2 (IL-2) deficiency occurring during cancer progression. Major surgery itself for cancer is known to induce lymphocytopenia in the postoperative period. Postoperative lymphocyte decrease in colorectal cancer can be prevented by preoperative administration of recombinant human (rhIL-2), indicating that it is possible to drive appropriately important host defence agents during critical events, such as major surgery. The aim of this study is to verify if recombinant human interleukin-2 (rhIL-2) administered preoperatively is able to prevent the lymphocyte decrease occurring after radical surgery in operable lung cancer. This phase II study included 40 patients with operable NSCLC screened as stage II or IIIA, randomized to receive rhIL-2, 9000000 IU subcutaneously twice daily for 3 days before surgery (treated group, 20 patients) or not (control group, 20 patients). At baseline, there were no significant differences in total lymphocyte number and lymphocyte subsets (T cell, T-helper, CD8+, natural killer, CD4/CD8 ratio) between groups. Postoperatively the control group showed a decrease in total lymphocyte count, T lymphocyte count, T-helper cell number and CD4/CD8 ratio, significant at the 14th postoperative day relative to baseline values. In contrast, in the rhIL-2 treated group, at the 3rd and at the 14th postoperative days, a significant increase was observed over both baseline and control group values of total lymphocyte count, T cells and T-helper cells. NK cell number increased significantly only over the control group. CD4/CD8 ratio was increased at the 14th postoperative day significantly over both baseline and control values. At pathological staging after surgery, four patients in the rhIL-2 group and four in the control group resulted in stage pIIIB; one patient in the rhIL-2 group resulted in stage IV (contralateral metastasis). Indeed, 15/20 rhIL-2 treated patients and 16/20 control patients were radically operated. After a 24-month follow-up, 12/20 rhIL 2 treated patients were alive and 8/15 radically operated were disease-free; 8/20 control patients were alive and 4/16 radically operated were disease-free. Toxicity was mild to moderate and easy manageable; treatment was suspended in one patient due to skin rash with hypotension grade II. The preoperative administration of rhIL-2 is feasible and prevents lymphocyte decrease occurring postoperatively after surgery for lung cancer. Further studies are required to assess the impact on survival. PMID- 9733055 TI - Two cases of intrapulmonary lymph node presenting as a peripheral nodular shadow: diagnostic differentiation from lung cancer. AB - We present two cases of intrapulmonary lymph node. The patients were a 44-year old woman and a 71-year-old man each with a small peripheral nodule in the lung. On computed tomography (CT) scans, both nodules were spiculated. Since histological diagnosis could not be obtained by bronchoscopic examination or CT guided needle biopsy, they underwent video-assisted thoracoscopic surgery. Histological examination of the resected material revealed that both nodules were composed of lymph node. Intrapulmonary lymph node has until recently been assigned no clinical significance; however, differential diagnosis of this lesion from lung cancers and other metastatic tumors is now clinically important. PMID- 9733056 TI - Immunohistochemical localization of placental leucine aminopeptidase/oxytocinase in normal human placental, fetal and adult tissues. AB - While oxytocinase is known to exist in pregnancy serum and placenta, the present study describes the expression of the mRNA for this enzyme in a wide variety of other human tissues. Northern blot analysis was used to detect the mRNA, with a probe derived from a cDNA for oxytocinase/placental leucine aminopeptidase (P LAP). Both the distribution and localization of immunoreactive oxytocinase/P-LAP protein have been determined immunohistochemically by use of an anti-P-LAP antibody in normal placental, fetal and adult tissues. In placental tissues, only syncytiotrophoblasts were stained positively. In both fetal and adult tissues, positive staining was obtained in vascular endothelial cells, gastrointestinal mucosal cells, epithelial cells of hepato-biliary, pancreato-biliary, bronchial alveolar and renal tubular systems as well as islet cells of pancreas and neurons in the central nervous systems. Sweat-gland cells, seminal vesicles and prostate gland in the adult, as well as adipocytes and skeletal muscle cells in the fetus were also stained. The widespread distribution of P-LAP suggests its involvement in a variety of physiological events not restricted to the regulation of the amounts of bioactive peptides such as arginine vasopressin (AVP) and oxytocin (OT) in pregnancy. The presence of P-LAP in syncytiotrophoblasts supports the idea that P-LAP in pregnancy serum is derived from the placenta. PMID- 9733057 TI - Effects of sustained hypoxaemia with 72 hours recovery on 11beta-hydroxysteroid dehydrogenase types 1 and 2 gene expression in near-term fetal sheep. AB - The study examined the effects of 8 h sustained hypoxaemia, with 72 h recovery, on the expression of 11beta-hydroxysteroid dehydrogenase (11beta-HSD) types 1 and 2 in near-term fetal sheep. Placental tissue and fetal liver and kidney were collected at Days 135-138 gestation 72 h after 8 h sustained hypoxaemia induced by lowering maternal inspired oxygen with (n = 9) and without (n = 6) metabolic acidosis or after 8 h normoxia (n = 6). In hypoxic fetuses with metabolic acidosis, a significant increase in the level of 11beta-HSD2 mRNA in the kidney compared with controls was correlated significantly with degree of associated fetal acidaemia, but there were no corresponding increases in the tissue level of 11beta-HSD2 activity. Hence, a time lag may exist between the mRNA and activity. Alternatively, the translation of 11beta-HSD2 mRNA may be inhibited. In contrast, levels of 11beta-HSD1 mRNA in the placenta and fetal liver were unchanged 72 h after sustained hypoxaemia. These results indicate that sustained fetal hypoxaemia with metabolic acidosis selectively up-regulates 11beta-HSD2 mRNA expression in the near-term fetal sheep kidney. This may be a re-bound effect at 72 h following an initial down-regulation as observed in a previous study. PMID- 9733058 TI - Follicular cells affect the fertilizability and developmental competency of bovine oocytes in vitro. AB - The present study examined the time-dependent effects of follicular cells on the fertilizability of oocytes and their subsequent development to blastocysts. The percentages of oocytes reaching the metaphase-II stage of maturation rose from 51.3% after 16 h of culture to 86.2% at 28 h (cumulus-intact oocytes; CIO) and, for the same time points, from 65.4% to 83.3% (corona-enclosed oocytes; CO) and 54.3% to 88.9% (denuded oocytes; DO), respectively. When DO were cultured for more than 24 h before insemination, fertilization rates were significantly lower compared with CIO and CO. The maximum rates of development to blastocysts were observed when the oocytes were cultured for 24 h in the CIO group (22.1%), 20 h in the CO group (19.7%) and 18 h in the DO group (9.2%), respectively. These results suggest that (i) the presence of cumulus cells or corona cells during maturation is not necessary for nuclear maturation of oocytes; (ii) the attachment of corona cells to the oocytes during maturation is important for the further development to the blastocyst stage, and (iii) the presence of attached cumulus and/or corona cells during maturation in vitro extends the maturation period required for further development to the blastocyst stage. PMID- 9733059 TI - Suppression of arousal by progesterone in fetal sheep. AB - The high rate of progesterone synthesis by the placenta in late gestation exposes the ovine fetus to high concentrations of progesterone and its metabolites that may affect activity of the fetal brain. The aim of this study was to determine the effect of inhibiting maternal progesterone synthesis on sleep-wake activity in fetal sheep. Fetal and maternal vascular catheters, a fetal tracheal catheter, and electrodes for recording fetal electrocortical (ECoG), electro-ocular (EOG) and nuchal muscle electromyographic (EMG) activity were implanted. At 128-131 days gestation, progesterone production was inhibited by an injection of trilostane (50 mg), a 3beta-hydroxysteroid dehydrogenase inhibitor. Vehicle solution or progesterone (3 mg h(-1)) was then infused into the ewe between 6 and 12 h after the trilostane treatment. Maternal progesterone concentrations were significantly reduced from 1-24 h after trilostane treatment (P < 0.05) when followed by vehicle infusion. Fetal breathing movements (FBM), EOG, nuchal muscle EMG, and behavioural arousal increased 12 h after trilostane treatment (P < 0.05). In contrast, there was no change in fetal arousal, EOG, EMG or FBM activities when progesterone was infused after the trilostane treatment. These findings show that progesterone can influence fetal behaviour, and indicates that normal progesterone production tonically suppresses arousal, or wakefulness in the fetus. PMID- 9733060 TI - A 39,X/40,XY true hermaphrodite mouse with normal ovarian function. AB - A cryptorchid mouse with a 39,X/40,XY chromosome constitution was identified among 414 offspring born in the departmental XO mouse breeding colony. This mouse had a small testis on the left, with no sign of spermatogenesis, and a normal ovary on the right with several corpora lutea. PMID- 9733061 TI - Fertilization of bovine oocytes grown in vitro. AB - Early antral follicles 0.5-0.7 mm in diameter were dissected from bovine ovaries, and oocyte-cumulus complexes with pieces of parietal granulosa (OCCGs) were then collected from the follicles. The OCCGs containing oocytes of 90-99 microm diameter (94.7+/-2.8 microm, n = 196) were selected and embedded in collagen gels and cultured for 14 days in TCM199 containing 10% fetal calf serum and 4 mM hypoxanthine. From cultured OCCGs, 144 surviving oocytes were recovered, of which 53 were granulosa cell-enclosed oocytes and 91 were denuded oocytes. The mean diameter of the surviving oocytes was 114.2+/-8.4 microm, significantly larger than that measured before culture (P < 0.05). The granulosa cell-enclosed oocytes and denuded oocytes were further cultured for maturation for 24 h. After culture, 72% (38/53) of the granulosa cell-enclosed oocytes and 59% (54/91) of the denuded oocytes showed normal morphology. These oocytes were then inseminated with bovine spermatozoa. After 29 h of insemination, all of the denuded oocytes had degenerated, while 32% (12/38) of the granulosa cell-enclosed oocytes showed normal morphology. Of the 12 oocytes, 5 were penetrated by spermatozoa, and 2 formed both male and female pronuclei. These results demonstrate for the first time that bovine oocytes grown in vitro acquire meiotic competence and can be penetrated by spermatozoa. PMID- 9733062 TI - Inducible nitric oxide synthase in the epithelial epididymal cells of the rat. AB - The expression of mRNA for inducible nitric oxide synthase (iNOS) in rat epithelial cells of epididymis was investigated with reverse transcription followed by polymerase chain reaction. Immunocytochemical reaction for iNOS was performed to confirm the enzyme's localization in the epididymal epithelium. Additionally, an indirect spectrophotometric method for nitric oxide (NO) determination was applied for measurement of nitrite production by cultured epididymal epithelial cells. Inducible NOS mRNA was detected in freshly isolated epithelial cells, in cultured cells without stimulation as well as in cultured cells after stimulation by lipopolysaccharide and interferon-gamma. Inducible NOS immunoreactivity was observed in the apical part of epithelial cells of epididymal sections and in the cytoplasm of cells in culture. Release of nitrite was observed in vitro in both the unstimulated and stimulated cells of caput (1.44+/-0.94 v. 4.37+/-2.42 microM) and cauda (0.69+/-1.21 v. 5.21+/-2.76 microM) epididymis (P < 0.001). To the best of our knowledge, this is the first study to demonstrate iNOS in the epididymal epithelial cells of the rat. Nitric oxide released by epididymal epithelial cells may act on cells and tissues located nearby. The results may help explain epididymal function: sperm storage, passage and maturation. Excessive epididymal NO production may also play a role in the inflammatory infertility of the male. PMID- 9733063 TI - The follicle-stimulating hormone beta-subunit gene of the common brushtail possum (Trichosurus vulpecula): analysis of cDNA sequence and expression. AB - Reverse transcription-PCR has been used to obtain a cDNA sequence from the follicle-stimulating hormone (FSH) beta-subunit gene of the Australian brushtail possum (Trichosurus vulpecula). Comparisons of the possum FSHbeta-mRNA coding region nucleotide sequence with that of six eutherian mammal homologues reveals a mean percent identity of 77.3% and 76.8% at the nucleotide and predicted amino acid-sequence levels respectively. Furthermore, the predicted amino acid sequence of the possum FSHbeta mature protein shows evolutionary conservation of twelve cysteine residues and two potential N-linked glycosylation sites. The protein lacks the CAGY motif present in most reported glycoprotein beta-subunit sequences. The translation termination codon and consensus polyadenylation sequence overlap, a feature observed in other mammalian FSHbeta genes. Northern hybridization of total RNA from adult female possum pituitary revealed three hybridizing transcripts of approximately 2.8, 1.2 and 0.5 kb which may arise from utilizing alternative polyadenylation signals. In situ hybridization localized the FSHbeta transcripts to a sub-population of anterior pituitary cells interpreted as being gonadotropes. In summary the results indicate considerable evolutionary conservation of the structure of the FSH beta-subunit gene between the marsupial and eutherian mammalian lineages. PMID- 9733064 TI - Arachidonic acid-induced acrosomal loss in the spermatozoa of a marsupial, the tammar wallaby (Macropus eugenii). AB - Tammar wallaby spermatozoa were induced to undergo acrosomal loss when incubated with arachidonic acid (AA). Ultrastructural examination indicated that the AA induced acrosomal loss occurred via multiple point fusions between the outer acrosomal membrane and the overlying plasma membrane. This form of acrosomal loss mimicked the physiological acrosome reaction (AR) seen in the sperm of eutherian mammals. The fusion event was limited to the acrosomal region of the plasma membrane and did not proceed past the peri-acrosomal ring. The entire acrosome was lost after AA treatment leaving no evidence of a persistent equatorial segment-like region. Ultrastructural evidence of AR-like membrane fusion was seen immediately on addition of 50 microg mL(-1) AA and a large proportion of sperm examined after five min incubation were in the late stages of membrane fusion. Longer-term incubation with AA had deleterious effects on wallaby sperm motility. It remains to be determined whether the AA-induced membrane fusion observed here indicates that AA is involved in the marsupial AR. However, pretreatment of sperm with the protein kinase C (PKC) inhibitor HMG significantly reduced AA-induced acrosomal loss suggesting that AA may have acted via PKC. If this is so, AA is probably physiologically significant and a novel pathway may be operating during AR induction in marsupials. PMID- 9733065 TI - Effect of maternal infusion of adrenocorticotrophin1-24 (ACTH1-24) on parturition in guinea-pigs near term. AB - While the mechanism of onset of labour in guinea-pigs is unknown, it has been suggested that administration of adrenocorticotrophin1-24 (ACTH1-24) near term induces labour. In order to verify this finding, guinea-pigs were fitted with indwelling carotid and jugular vascular cannulae. ACTH1-24 (30 microg h(-1) for four hours, n = 9) or vehicle (n = 5) was infused intravenously on Day 64 (term is 68 days). ACTH1-24 had no effect on gestational length (68.4+/-1.0 days, n = 6 v. control, 69.6+/-0.3 days, n = 5, P = 0.8). Symphysial width, fetal weight, number and viability were similar in both groups (all P > 0.1). Infusion of ACTH1 24 increased maternal ACTH concentrations from <1.8 pmol L(-1) to 34+/-6 pmol L( 1) (n = 6, P < 0.01) while fetal ACTH concentrations remained undetectable (n = 6). Infusion of ACTH1-24 increased cortisol concentrations in maternal plasma from 8.3+/-0.6 mmol L(-1) to 15.8+/-0.8 mmol L(-1) (n = 6, P < 0.001) but had no effect on concentrations of 13,14-dihydro-15-keto-PGF2alpha (P = 0.8). It is concluded that (1) maternal infusion of ACTH1-24 at the dosage used does not induce labour in guinea-pigs, and (2) ACTH1-24 does not cross the placenta. PMID- 9733066 TI - Early onset of parturition induced by acute alcohol exposure in C57BL/6J mice: role of uterine PGE and PGF2alpha. AB - These studies were designed to determine the effect of acute alcohol treatment on gestational length and to probe for a mechanism underlying alcohol-induced early onset of parturition (EOP) in mice. Experiment 1: alcohol increases the incidence of EOP. Pregnant C57BL/6J mice were given alcohol (0, 4, 5 or 6 g kg(-1), i.g.) on Gestational Day (GD) 10, 15, 16, 17 or 18. Deliveries were monitored every 6 h from GD 18. Results indicated that 6 g kg(-1) alcohol treatment on GD 17 or 18 increased the incidence of EOP. Experiment 2: prostaglandins (PGs) play roles in parturition. The purpose of Experiment 2 was to determine whether PGs mediate alcohol-induced EOP in mice. The results indicated that pretreatment on GD 17 with aspirin, a prostaglandin synthesis inhibitor, prevented alcohol-induced EOP. These data suggest that alcohol-induced EOP in mice may be mediated by PGs. Experiment 3: PGs are influenced by alcohol and are triggers of labour. Experiment 3 measured uterine PGs associated with the onset of alcohol-induced EOP in mice. Alcohol increased uterine PGE and PGF2alpha, with PGE levels higher than control before labour, and elevated PGF2alpha levels correlating with labour. Changes in gestational length have important implications for pregnancy outcome, as well as for normal fetal growth and development. PMID- 9733067 TI - Differential expression pattern of inhibin alpha and betaA subunits in the ovaries of postnatal and prepubertal lambs. AB - The aim of the present study was to characterize gene and protein expression of follistatin, inhibin alpha (alpha) and inhibin betaA (betaA) subunits in the ovaries of postnatal (3-week-old) and prepubertal (14 and 20-25-week-old) lambs. Northern blot analysis revealed the presence of two alpha and two betaA mRNAs. In postnatal ovary the a 1.2-kb transcript was abundant, its amount gradually falling, while the 2.0-kb mRNA increased and became a major band at 20-25 weeks. Both betaA mRNAs, 4.5 kb and 6.0-7.5 kb, were weakly expressed in postnatal ovary, but whereas the 4.5-kb mRNA expression remained at a low level, that of the 6.0-7.5 kb mRNA increased about five fold in prepubertal ovary. The ratio of total alpha mRNAs to the dominant betaA form (6.0-7.5 kb) varied from 1.27 at 3 weeks to 0.33 at 20-25 weeks of age. One major follistatin mRNA of 2.5-3.6 kb was recognized and was constitutively expressed during ovarian growth. Several molecular-mass forms of alpha and betaA subunits with different compositions were seen in prepubertal compared with postnatal ovaries, the latter exhibiting more active follicular growth. In summary, ovine ovaries undergo distinct changes early in life, both morphological and functional, and show a changing pattern of inhibin subunit expression. PMID- 9733068 TI - Up-regulation of protein kinase C in regenerating optic nerve fibers of goldfish: immunohistochemistry and kinase activity assay. AB - Protein kinase C (PKC) activation has been associated with synaptic plasticity in many projections, and manipulating PKC in the retinotectal projection strongly affects the activity-driven sharpening of the retinotopic map. This study examined levels of PKC in the regenerating retinotectal projection via immunostaining and assay of activity. A polyclonal antibody to the conserved C2 (Ca2+ binding) domain of classical PKC isozymes (anti-panPKC) recognized a single band at 79-80 kD on Western blots of goldfish brain. It stained one class of retinal bipolar cells and the ganglion cells in normal retina, as shown previously. Strong staining was not present in the optic fiber layer of retina or in optic nerve, optic tract, or terminal zone in tectum, with the exception of a single fascicle of optic nerve fibers that by their location and by L1 (E587) staining were identified as those arising from newly added ganglion cells at the retinal margin. Normal tectal sections showed dark staining of a subclass of type XIV neuron with somas at the top of the periventricular layer and an apical dendrite ascending to stratum opticum. In regenerating retina, swollen ganglion cells stained darkly and stained axons were seen in the optic fiber layer. In regenerating optic nerve (2-11 weeks postcrush), all fascicles of optic fibers stained darkly for both PKC and L1(E587). At 5 weeks postcrush, PKC staining could also be seen in the medial and lateral optic tracts and stratum opticum at the front half of the tectum and very lightly over the terminal zones. PKC activity was measured in homogenized tissues dissected from a series of fish with unilateral nerve crush from 1 to 5 weeks previously. Activity levels stimulated by phorbols and Ca2+ were measured by phosphorylation of a specific peptide and referred to levels measured in the opposite control side. Regeneration did not increase overall PKC activity in retina or tectum, but in optic nerve there was an 80% rise after the first week. The increased activity verifies that the increased staining in nerve represented an up-regulation of functional PKC during nerve regeneration. PMID- 9733069 TI - Pituitary adenylate cyclase activating polypeptide (PACAP) expression in sympathetic preganglionic projection neurons to the superior cervical ganglion. AB - Pituitary adenylate cyclase activating polypeptides (PACAP27 and PACAP38) are members of the VIP/secretin/glucagon family of peptides and have diverse neuroregulatory effects in sympathoadrenal cell development and function. PACAP peptides regulate rat superior cervical ganglion (SCG) neuron catecholamine and neuropeptide Y content and secretion, and promote sympathoneuroblast survival through activation of specific PACAP1 receptor isoforms. In examining the potential sources of PACAP regulating the SCG, PACAP expression was identified in rat preganglionic neurons in the intermediolateral cell column (IML) of the thoracic spinal cord which provide primary afferent projections to this sympathetic ganglion. Thoracic spinal cord segments (T1-4) contained approximately 17 pmol PACAP38 immunoreactivity/g tissue wet weight. Reverse transcription polymerase chain reaction of cDNA from microdissected thoracic spinal cord using primers specific for rat neuronal proPACAP identified proPACAP mRNA expression in the IML; the results correlated with neurons labeled for proPACAP mRNA by in situ hybridization histochemistry and implicated PACAP biosynthesis in IML neurons. To demonstrate directly proPACAP transcript expression in preganglionic projection neurons to the SCG, the ganglion was decentralized and the sympathetic trunk immersed in fluorogold to identify sympathetic preganglionic neurons by retrograde labeling. Cryosections of spinal cord segments containing preganglionic neuron fluorogold labeled neurons were processed subsequently for in situ hybridization histochemical localization of proPACAP mRNA using a digoxigenin-labeled riboprobe; IML neurons were examined for fluorogold and digoxigenin/alkaline phosphatase product dual labeling. More than half of the preganglionic projection neurons to the SCG expressed PACAP mRNA, consistent with the postulate that PACAP peptides released from a subpopulation of thoracic IML preganglionic neurons may be physiological anterograde modulators of sympathetic SCG function. PMID- 9733070 TI - Disconnected mutants show disruption to the central projections of proprioceptive neurons in Drosophila melanogaster. AB - We used a P[GAL4] enhancer-trap line, C161, in conjunction with the UAS-lacZ reporter construct to visualize the central projections of a defined set of thoracic and abdominal sensory neurons in a disconnected (disco) mutant background. The results show defects in the organization of sensory axons in the larval and adult central nervous system. The defects are indicative of problems with axon growth and development and include (a) poor axon fasciculation, (b) aberrant axon growth, (c) excessive terminal branching, and (d) ectopic innervation. Sensory neuron identity appears to be normal. The defects are comparable to those previously described for larval photoreceptor and adult retinular cells in disco mutants and extend the known effects of this mutation. Reduced larval and adult viability are likely to result from locomotory defects related to the disruption of the sensory system. PMID- 9733071 TI - Continuous adult development of multiple innervation in toadfish sonic muscle. AB - The sonic muscle of the oyster toadfish Opsanus tau produces unfused contractions at over 200 Hz for mating call production, requiring extreme muscle fiber synchronization. This multiply innervated muscle is sexually dimorphic and grows for life by fiber proliferation and hypertrophy. Previous descriptions of its multiple innervation did not consider fish size or sex. We examined neuromuscular junction (NMJ) development in adult fish of both sexes between 123 and 343 mm in total length (24.7790 g in mass). The NMJ was a tubelike trough that varied in length from 8 to 178 microm. Troughs were usually straight, although some consisted of consecutive ovals and some were branched. Median length of NMJs increased linearly with fish length (r2=.40; p=.002) from 58 to 75 microm. Modal lengths were mostly between 50 and 60 microm and did not increase ontogenetically, indicating that the median increase was caused by a greater number of large junctions in older fish. Median interval between NMJs (measured from the beginning of one junction to the next) ranged from 92 to 116 microm and did not vary with fish size (r2=.06; p=.285). Considering muscle fiber elongation, the data indicate an increase from 60 to 140 NMJs per fiber during fish growth. There were no sexual differences in NMJ length or spacing. In view of the slow conduction velocity of sonic muscle fibers, the addition of new NMJs and the relatively constant distance between them supports rapid and synchronized contraction necessary for sound production in both sexes. PMID- 9733072 TI - Expression and neuropeptidergic characterization of estrogen receptors (ERalpha and ERbeta) throughout the rat brain: anatomical evidence of distinct roles of each subtype. AB - The recent cloning of a second estrogen receptor (ER) provided a new tool to investigate and clarify how estrogens are capable of communicating with the brain and influence gene expression and neural function. The purpose of the present study was to define the neuroanatomical organization of each receptor subtype using a side-by-side approach and to characterize the cellular population (s) expressing the ERbeta transcript in the endocrine hypothalamus using immunohistochemistry combined with in situ hybridization. Axonal transport inhibition was accomplished to cause neuropeptide accumulation into the cytoplasm and thus facilitate the detection of all positive luteinizing hormone-releasing hormone (LHRH), corticotropin-releasing factor (CRF), vasopressin (AVP), oxytocin (OT), gastrin-related peptide (GRP), and enkephalin (ENK) neurons. The genes encoding either ERalpha or -beta were expressed in numerous limbic-associated structures, and fine differences were found in terms of intensity and positive signal. Such phenomenon is best represented by the bed nucleus of the stria terminalis (BnST) and preoptic area/anterior hypothalamus, where the expression pattern of both transcripts differed across subnuclei. The novel ER was also found to be expressed quite exclusively in other hypothalamic nuclei, including the supraoptic (SON) and selective compartments (magnocellular and autonomic divisions) of the paraventricular nucleus (PVN). A high percentage of the ERbeta expressing neurons located in the ventro- and dorsomedial PVN are of OT type; 40% of the OT-ir cells forming the medial magnocellular and ventromedial parvocellular PVN showed a clear hybridization signal for ERbeta mRNA, whereas a lower percentage (15-20%) of OT neurons were positive in the caudal parvocellular PVN and no double-labeled cells were found in the rostral PVN and other regions of the brain with the exception of the SON. Very few AVP-ir neurons expressing ERbeta transcript were found throughout the rat brain, although the medial PVN displayed some scattered double-labeled cells (<5%). Quite interestingly, the large majority of the ERbeta-positive cells in the caudal PVN were colocalized within CRF-ir perikarya. Indeed, more than 60-80% of the CRF-containing cells located in the caudolateral division of the parvocellular PVN exhibited a positive hybridization signal for ERbeta mRNA, whereas very few (<5%) neuroendocrine CRF-ir parvocellular neurons of the medial PVN expressed the gene encoding ERbeta. A small percentage of ERbeta-expressing cells in the dorsocaudal and ventromedial zones of the parvocellular PVN were also ENK positive. The ventral zone of the medial parvocellular PVN also displayed GRP-ir neurons, but no convincing hybridization signal for ERbeta was detected in this neuronal population. Finally, as previously described for the gene encoding the classic ER, LHRH neurons of both intact and colchicine-pretreated animals did not express the novel estrogen receptor. This study shows a differential pattern of expression of both receptors in the brain of intact rats and that ERbeta is expressed at various levels in distinct neuropeptidergic populations, including OT, CRF, and ENK. The influence of estrogen in mediating genomic and neuronal responses may therefore take place within these specific cellular groups in the brains of cycling as well as intact male mammals. PMID- 9733074 TI - Electrical membrane properties of trapezoid body neurons in the rat auditory brain stem are preserved in organotypic slice cultures. AB - The medial nucleus of the trapezoid body (MNTB) is a conspicuous structure in the mammalian auditory brain stem. It is a major component of the superior olivary complex and is involved in sound localization. Recently, organotypic slice culture preparations of the superior olivary complex were introduced to investigate the development of inhibitory and excitatory projections (Sanes and Hafidi, 1996; Lohmann et al., 1998). In the present article, we further assessed the organotypicity of our culture system (Lohmann et al., 1998) and examined electrical membrane properties of MNTB neurons expressed under culture conditions. To do so, MNTB neurons from early postnatal rats (P3-5) were studied after 3-6 days in vitro (DIV) by whole-cell patch-clamp recordings. Their mean resting potential was -59 mV, the input resistance averaged 171 Momega, and the average time constant was 3 ms. Four types of voltage-activated conductances were observed in voltage-clamp recordings. All cells expressed a tetrodotoxin (TTX) sensitive sodium current. Two types of potassium currents could be characterized: a tetraethylammonium (TEA) -sensitive and a 4-aminopyridine (4-AP)-sensitive conductance, both of which are composed of a transient and a sustained component. Finally, an inwardly rectifying current, activated by hyperpolarizing voltage steps, was found. In current-clamp recordings, depolarizing current pulses typically elicited a single action potential. In the presence of 4-AP, however, these current pulses induced a train of action potentials. The duration of action potentials was increased by 4-AP and the afterhyperpolarization was reduced. Hyperpolarizing current injections induced a "sag" in the membrane potential, indicating the influence of an inwardly rectifying current. Our results demonstrate that MNTB neurons in slice cultures have electrical membrane properties comparable to those of their counterparts in acute slices. PMID- 9733073 TI - Mechanisms of insulin-like growth factor regulation of programmed cell death of developing avian motoneurons. AB - During development of the avian neuromuscular system, lumbar spinal motoneurons (MNs) innervate their muscle targets in the hindlimb coincident with the onset and progression of MN programmed cell death (PCD). Paralysis (activity blockade) of embryos during this period rescues large numbers of MNs from PCD. Because activity blockade also results in enhanced axonal branching and increased numbers of neuromuscular synapses, it has been postulated that following activity blockade, increased numbers of MNs can gain access to muscle-derived trophic agents that prevent PCD. An assumption of the access hypothesis of MN PCD is the presence of an activity-dependent, muscle-derived sprouting or branching agent. Several previous studies of sprouting in the rodent neuromuscular system indicate that insulin-like growth factors (IGFs) are candidates for such a sprouting factor. Accordingly, in the present study we have begun to test whether the IGFs may play a similar role in the developing avian neuromuscular system. Evidence in support of this idea includes the following: (a) IGFs promote MN survival in vivo but not in vitro; (b) neutralizing antibodies against IGFs reduce MN survival in vivo; (c) both in vitro and in vivo, IGFs increase neurite growth, branching, and synapse formation; (d) activity blockade increases the expression of IGF-1 and IGF-2 mRNA in skeletal muscles in vivo; (e) in vivo treatment of paralyzed embryos with IGF binding proteins (IGF-BPs) that interfere with the actions of endogenous IGFs reduce MN survival, axon branching, and synapse formation; (f) treatment of control embryos in vivo with IGF-BPs also reduces synapse formation; and (g) treatment with IGF-1 prior to the major period of cell death (i.e., on embryonic day 6) increases subsequent synapse formation and MN survival and potentiates the survival-promoting actions of brain-derived neurotrophic factor (BDNF) and glial cell line-derived neurotrophic factor (GDNF) administered during the subsequent 4- to 5-day period of PCD. Collectively, these data provide new evidence consistent with the role of the IGFs as activity-dependent, muscle derived agents that play a role in regulating MN survival in the avian embryo. PMID- 9733075 TI - Photoperiod regulates neuronal bromodeoxyuridine labeling in the brain of a seasonally breeding mammal. AB - Seasonal changes in vertebrate brain function are pervasive, but annual cycles in the rates of neuronal incorporation are established only in songbirds. Although cell division continues in the subependymal and hippocampal subgranular zones of adult rodents, there exists no parallel evidence that seasonal plasticity in mammals extends to changes in neuronal or glial number. We examined the effect of photoperiod on incorporation of new neurons in the brain of the adult golden hamster, a long-day breeder. We administered the cell birth marker 5' bromodeoxyuridine (BrdU) to males which had either been maintained in long days, transferred to short days for 10 weeks, or moved acutely from long to short or short to long days. The number of cells in specific brain regions immunoreactive (ir) for this thymidine analog was determined 7 weeks later. The number of BrdU ir cells in the dentate gyrus and subependymal zone increased twofold in short days. Transfer between photoperiods 10 days before the BrdU injections produced intermediate numbers of BrdU-labeled cells in the dentate gyrus, but was as effective as long-term photoperiodic exposure in the subependymal zone. Photoperiod also had similar effects in the hypothalamus and cingulate/retrosplenial cortex, but not in the central gray or preoptic area. Double-label immunocytochemistry indicated that very few of the BrdU-ir cells were glia, but that a majority had neuronal phenotype. In the subependymal zone, short days significantly increased the number of BrdU-labeled neurons. We did not detect significant effects of photoperiod on the volume of either the granule cell layer of the hippocampus or the dentate gyrus as a whole. We conclude that short day lengths increase neuronal birth and/or survival in several brain regions of adult hamsters. PMID- 9733076 TI - Neural correlates of singing behavior in male zebra finches (Taeniopygia guttata). AB - This study examined the relationship between the volumes of four song control nuclei: the high vocal center (HVC), the lateral part of the magnocellular nucleus of the anterior neostriatum (IMAN), Area X, and the robust nucleus of the archistriatum (RA), as well as syrinx mass, with several measures of song output and song complexity in male zebra finches (Taeniopygia guttata). Male zebra finches' songs were recorded in standardized recording sessions. The syrinx and brain were subsequently collected from each bird. Volumes of the song control nuclei were reconstructed by measuring the cross-sectional area of serial sections. Syrinx mass was positively correlated with RA volume. The volume of IMAN was negatively related to element repertoire size and the number of elements per phrase. We found no other correlations between brain and behavioral measures. This study, combined with others, indicates that the evidence for a general relationship among songbirds between HVC volume and song complexity is equivocal. There are clear species differences in this brain-behavior correlation. PMID- 9733078 TI - Inspiratory muscle activity during bird song. AB - The apparently continuous flow of bird song is in reality punctuated by brief periods of silence during which there are short inspirations called minibreaths. To determine whether these minibreaths are accompanied, and thus perhaps caused, by activity in inspiratory muscles, electromyographic (EMG) activity was recorded in M. scalenus in zebra finches and in M. scalenus and Mm. levatores costarum in cowbirds, together with EMGs from the abdominal expiratory muscles, air sac pressure and tracheal airflow. EMG activity in Mm. scalenus and levatores costarum consistently preceded the onset of negative air sac pressure by approximately 11 ms during both quiet respiration and singing in both species. The electrical activity of these two muscles was very similar. Compared with during quiet respiration, the amplitude of inspiratory muscle EMG during singing was increased between five- and 12-fold and its duration was decreased from >200 ms to on average 41 ms during minibreaths, again for both species, but inspiratory muscle activity did not overlap with that of the expiratory muscles. Thus, there was no indication that the inspiratory muscles acted either to shorten the duration of expiration or to reduce the expiratory effort as might occur if both expiratory and inspiratory muscles were simultaneously active. Inspiratory and expiratory muscle activities were highly stereotyped during song to the extent that together, they defined the temporal pattern of the songs and song types of individual birds. PMID- 9733077 TI - Ectopic expression of p27Kip1 in oligodendrocyte progenitor cells results in cell cycle growth arrest. AB - Oligodendrocyte differentiation is a complex process believed to be controlled by an intrinsic mechanism associated with cell-cycle arrest. Recently, the cell cycle inhibitor protein p27 Kip1 has been proposed as a key element in causing growth arrest of oligodendrocyte precursor cells. To investigate the effects of p27 upon oligodendrocyte cell development, we have introduced the p27 cDNA in oligodendrocyte progenitor cells using an adenovirus vector. Progenitor cells normally express low levels of p27. After adenoviral infection and p27 overexpression, progenitor cells were able to undergo cell-cycle arrest, even in the presence of strong mitogens. The effects of p27 were shown to be directly upon cyclin-dependent kinase-2 (CDK2), the protein kinase complex responsible for G1/S transition, as immunodepletion of oligodendrocyte extracts of p27 protein resulted in the activation of CDK2 activity. However, cells that became growth arrested owing to infection with p27 adenovirus did not display conventional oligodendrocyte differentiation markers, such as O4 or O1. Taken together, these data provide mechanistic evidence indicating that p27 is primarily involved in oligodendroglial progenitor proliferation by inhibiting CDK2 activity and inducing oligodendrocyte cell-cycle arrest. PMID- 9733079 TI - Ontogenetic alteration in peptidergic expression within a stable neuronal population in lobster stomatogastric nervous system. AB - In the adult lobster, Homarus gammarus, the stomatogastric ganglion (STG) contains two well-defined motor pattern generating networks that receive numerous modulatory peptidergic inputs from anterior ganglia. We are studying the appearance of extrinsic peptidergic inputs to these networks during ontogenesis. Neuron counts indicate that as early as 20% of development (E20) the STG neuronal population is quantitatively established. By using immunocytochemical detection of 5-bromo-2'-deoxyuridine incorporation, we found no immunopositive cells in the STG by E70. We concluded that the STG neuronal population remains quantitatively stable from mid-embryonic life until adulthood. We then investigated the ontogeny of FLRFamide- and proctolin-like peptides in the stomatogastric nervous system, from their first appearance until adulthood by using whole mount immunocytochemistry. Numerous FLRFamide-like-immunoreactive STG neuropilar ramifications were observable as early as E45 and remain thereafter. From E50 to the first larval stage, one to three STG somata stained, while somatic staining was not observed in larval stage II and subsequent stages. From E50 and thereafter, the STG neuropilar area was immunopositive for proctolin. One to two proctolinergic somata were detected in the STG of the three larval stages but were not seen in embryos, the post-larval stage or in adults. Thus, peptidergic inputs to the STG are present from mid-embryonic life. Moreover, whereas in the adult, STG neurons only contain glutamate or acetylcholine, some neurons transiently express peptidergic phenotypes during development. Although this system expresses an ontogenetic peptidergic plasticity, the STG neurons produce a single stable embryonic-larval motor output (Casasnovas and Meyrand [1995] J. Neurosci. 15:5703-5718). PMID- 9733080 TI - Quantitative analysis of cerebellar lobulation in normal and agranular rats. AB - Cerebellar pattern formation was investigated in rats treated with DNA modifying agents. Animals were subjected to combinations of daily injections of methylazoxymethanol acetate (MAM) for the last 6 days gestation and/or localised X-irradiation of the hindbrain on postnatal days 1 and 5 (P1 and P5). Animals were analysed on embryonic day 18 (E18), P0, P3, P7, and P14. Five parameters of the cerebellum were recorded from midsagittal sections: the number of primary lobules; the thickness of the external germinal layer (EGL); the density of cells in the internal granule cell layer (IGL) region; and the midsagittal area and perimeter. In addition, the laterolateral cerebellar distance was calculated. The data demonstrate that pre- and postnatal reduction of the EGL results in reduced cerebellar growth and folding. Cessation of the treatment at birth results in a recovery and eventual overproduction of EGL, but cerebellar growth and the development of fissures lags behind that of normal rats. Pre- and postnatal destruction of the EGL severely limited cerebellar growth and fissuration, and the cerebella contained only five primary lobules at P14. Rats subjected to postnatal X-irradiation alone had a similar low density of granule cells relative to those treated with a combination of prenatal MAM injections and postnatal X irradiation, and yet the cerebella contained deeper fissures and more lobules (nine at P14). The data indicate that there are two phases of cerebellar folding: the establishment of five lobules that arise independent of granule cell production, and the granule cell-dependent expansion and partitioning of these five principal lobules during postnatal development. We propose that the lack of correlation between the severity of the granule cell loss and degree of lobulation in agranular rats indicates that granule cells exert an inductive influence over lobulation that is in part independent of the forces generated by their production and differentiation. PMID- 9733081 TI - Galanin-R1 receptor in anterior and mid-hypothalamus: distribution and regulation. AB - The distribution and regulation of galanin-R1 receptor (GAL-R1-R) mRNA has been studied in the anterior and mid-diencephalon by using in situ hybridization. Moreover, possible colocalization of GAL-R1-R mRNA and prepro-galanin or vasopressin mRNAs has been analyzed at the cellular level using double in situ hybridization methodology. Many nuclei in the hypothalamus expressed GAL-R1-R mRNA, including the paraventricular nucleus (PVN) and the supraoptic nucleus (SON). Strong expression was also seen in the same sections in various areas outside of the diencephalon. The distribution patterns are similar to those described in earlier studies. Double labeling experiments showed GAL-R1-R mRNA in vasopressin neurons in the PVN and SON. Moreover, GAL-R1-R mRNA and prepro galanin mRNA were colocalized in several hypothalamic nuclei. GAL-R1-R mRNA levels showed a high degree of plasticity. Thus, salt loading resulted in a marked increase in GAL-R1-R mRNA levels in the PVN and SON and a moderate decrease was seen during lactation. In contrast, hypophysectomy caused a decrease in GAL-R1-R mRNA levels. Differential effects of colchicine were recorded with a decrease of GAL-R1-R mRNA in the magnocellular hypothalamic neurons. After salt loading or during lactation, GAL-R1-R mRNA and prepro-galanin mRNA were regulated in parallel, whereas their levels changed in opposite directions after hypophysectomy and colchicine injection. In conclusion, GAL-R1-Rs are present in several hypothalamic nuclei, partly in neurons synthesizing galanin. The receptors are regulated in a specific fashion in the various nuclei, depending on the stimulus applied. The results suggest that the effect of galanin in the hypothalamus partly depends on the state of receptor expression. PMID- 9733082 TI - Chemical anatomy of excitatory endings in the dorsal cochlear nucleus of the rat: differential synaptic distribution of aspartate aminotransferase, glutamate, and vesicular zinc. AB - In order to identify cytochemical traits relevant to understanding excitatory neurotransmission in brainstem auditory nuclei, we have analyzed in the dorsal cochlear nucleus the synaptic distribution of aspartate aminotransferase, glutamate, and vesicular zinc, three molecules probably involved in different steps of excitatory glutamatergic signaling. High levels of glutamate immunolabeling were found in three classes of synaptic endings in the dorsal cochlear nucleus, as determined by quantitation of immunogold labeling. The first type included auditory nerve endings, the second were granule cell endings in the molecular layer, and the third very large endings, better described as "mossy." This finding points to a neurotransmitter role for glutamate in at least three synaptic populations in the dorsal cochlear nucleus. The same three types of endings enriched in glutamate immunoreactivity also contained histochemically detectable levels of aspartate aminotransferase activity, suggesting that this enzyme may be involved in the synaptic handling of glutamate in excitatory endings in the dorsal cochlear nucleus. There was also extrasynaptic localization of the enzyme. Zinc ions were localized exclusively in granule cell endings, as determined by a Danscher-selenite method, suggesting that this ion is involved in the operation of granule cell synapses in the dorsal cochlear nucleus. PMID- 9733083 TI - Apoptosis of undifferentiated progenitors and granule cell precursors in the postnatal human cerebellar cortex correlates with expression of BCL-2, ICE, and CPP32 proteins. AB - Naturally occurring apoptotic cells have been demonstrated in the postnatal cerebellum of rodents (Wood et al. [1993] Neuron 11:621-632; Krueger et al. [1995] J. Neurosci. 15:3366-3374). The nature of these cells differs among species: they are considered to be granule cells in mouse and astrocytes in rat. We labeled proliferating and apoptotic cells in the postnatal human cerebellar cortex by using antibodies against the Ki-67/proliferating cell nuclear antigen and the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling method for fragmented DNA. We also immunocytochemically detected some proteins encoded by genes modulating apoptosis and specific markers of neuronal/glial differentiation. Proliferating cells were observed from birth to 4 months, representing 31-35% of cells within the external granular layer (EGL). Apoptotic cells were detected during the first 3 months and corresponded to 5-7% of EGL cells. Much lower percentages were calculated in other cortical layers and white matter. The balance between proliferation and apoptosis was quantitatively favorable to the latter during the first postnatal week. Expression of BCL-2, CPP32, and interleukin-1beta-converting enzyme (ICE) proteins was spatially and developmentally regulated in parallel with apoptosis. Apoptotic cells were often CPP32/ICE immunoreactive but negative for BCL-2. Some apoptotic cells were positive for vimentin and, less frequently, for alpha-internexin or type-III beta tubulin, but never expressed the glial fibrillary acidic protein. This study demonstrates that apoptosis is a significant phenomenon in early postnatal development of human cerebellar cortex and shares some of the regulatory mechanisms described in other vertebrates. PMID- 9733084 TI - ORL-1 and mu opioid receptor antisera label different fibers in areas involved in pain processing. AB - Mu opioid receptors (MOR) mediate the analgesic effects of opioid drugs such as morphine. The opioid receptor-like (ORL-1) receptor is structurally related to opioid receptors and the ORL-1 receptor agonist, orphanin FQ/nociceptin, induces analgesia at the spinal level, but appears to recruit different circuitry than that used by mu opioids. When administered intracerebroventricularly, orphanin FQ/nociceptin produces hyperalgesia and/or reverses opioid analgesia. The functionally distinct actions elicited by MOR and ORL-1 receptors, which activate similar intracellular signaling systems and show similar regional distributions, could be explained by their differential cellular localization. By using double label immunohistochemistry and confocal microscopy, the present study investigates the distribution of MOR and ORL-1 receptors in regions of the rat nervous system that are involved with nociceptive processing. In general co localization of MOR and ORL-1 receptor immunoreactivity was not observed in either perikarya or neuropil in the dorsal root ganglia, nor in the Lissauer's tract and superficial laminae of the spinal cord. Likewise, there was no evidence for co-localization of these receptors within the periaqueductal gray, the nucleus raphe magnus, the gigantocellular reticular nucleus, and the nucleus of the solitary tract. These observations indicate that MOR and ORL-1 receptors are expressed predominantly on different fiber systems in these regions. This differential distribution is consistent with the distinct pharmacology of ORL-1 and MOR receptor agonists and suggests that the antisera to MOR and ORL-1 receptors may provide useful markers for further investigations of analgesic and counteranalgesic pathways modulating pain perception. PMID- 9733085 TI - Corticostriatal connections of the superior temporal region in rhesus monkeys. AB - Corticostriatal connections of auditory areas within the supratemporal plane and in rostral and caudal portions of the superior temporal gyrus were studied by the autoradiographic anterograde tracing technique. The results show that the primary auditory cortex has limited projections to the caudoventral putamen and to the tail of the caudate nucleus. In contrast, the second auditory area within the circular sulcus has connections to the rostral and the caudal putamen and to the body of the caudate nucleus and the tail. The association areas of the superior temporal gyrus collectively have widespread corticostriatal projections characterized by differential topographic distributions. The rostral part of the gyrus projects to ventral portions of the head of the caudate nucleus and of the body and to the tail. In addition, there are connections to rostroventral and caudoventral portions of the putamen. The mid-portion of the gyrus projects to similar striatal regions, but the connections to the head of the caudate nucleus are less extensive. Compared with the rostral and middle parts of the superior temporal gyrus, the caudal portion has little connectivity to the tail of the caudate nucleus. It projects more dorsally within the head and the body and also more dorsally within the caudal putamen. These differential patterns of corticostriatal connectivity are consistent with functional specialization at the cortical level. PMID- 9733086 TI - Sympathetic innervation of the upper and lower regions of the uterus and cervix in the rat have different origins and routes. AB - The origins and routes of the postganglionic sympathetic nerve supply to the upper and lower uterus and to the cervix were investigated in the rat by using denervation procedures combined with immunohistochemistry and retrograde tracing. The sympathetic nerve fibers of the upper part of the uterus arise from the ovarian plexus nerve. They mainly originate (90%) from neurons of the suprarenal ganglia (SRG) and of the T10 to L3 ganglia of the paravertebral sympathetic chain. Fluoro-Gold injections into different regions of the upper uterus showed that the SRG neurons mainly provide innervation to the tubal extremity (52%) rather than to the uterine portion below this area (26%). Very few neurons of the celiac ganglion or the aorticorenal ganglia participated in this innervation. Most of the sympathetic innervation of the lower uterus and the cervix (90%) originates from neurons of the paravertebral ganglia T13 to S2, principally at the L2-L4 levels. By using immunocytochemistry, we show that very few tyrosine hydroxylase-positive neurons of the pelvic plexus project to these areas, where they represent only 3% of the sympathetic nerve supply. Again, very few neurons of the inferior mesenteric ganglion (IMG) supply the lower uterus and the cervix. The comparison between retrograde tracing experiments in intact animals and after the removal of the IMG shows that very few sympathetic postganglionic axons from the paravertebral chain pass through the IMG to reach the lower uterus and the cervix. In contrast, these axons mainly project to splanchnic nerves bypassing the IMG to connect with the hypogastric nerves. In addition, some axons supplying the lower uterus follow the superior vesical arteries and then reach the organ. Taken together, these results show that the upper region of the uterus receives a sympathetic innervation that is different in origin and route from that of the lower uterus and the cervix. Such a marked region-specific innervation suggests that nerve control of the myometrial activity may be functionally different between the oviduct and the cervical ends of the uterus. PMID- 9733087 TI - Transforming growth factor alpha with insulin stimulates cell proliferation in vivo in adult rat vestibular sensory epithelium. AB - Hair cells, the sensory receptors of the mammalian inner ear, have long been thought to be produced only during embryogenesis, and postnatal hair cell loss is considered to be irreversible and is associated with permanent hearing and balance deficits. Little is known about the factors that regulate hair cell genesis and differentiation. The mitogenic effects of insulin and transforming growth factor alpha (TGFalpha) were assayed in vivo in normal and drug-damaged rat inner ear. Tritiated thymidine and autoradiographic techniques were used to identify cells synthesizing DNA. Simultaneous infusion of TGFalpha and insulin directly into the inner ear of adult rats stimulated DNA synthesis in the vestibular sensory receptor epithelium. New supporting cells and putative new hair cells were produced. Infusion of insulin alone or TGFalpha alone failed to stimulate significant DNA synthesis. These results suggest that exogenous growth factors may have utility for therapeutic treatment of hearing and balance disorders in vivo. PMID- 9733088 TI - Mitochondrial Ca2+ transients in cardiac myocytes during the excitation contraction cycle: effects of pacing and hormonal stimulation. AB - Using laser scanning confocal microscopy, our objective was to measure mitochondrial, nuclear, and cytosolic free ionized Ca2+ in adult rabbit cardiac myocytes loaded with Ca2+-indicating fluorophores. When myocytes were loaded with Fluo 3 at 37 degrees C, the fluorophore was loaded extensively into the cytosol and nucleus, but poorly into mitochondria, and Fluo 3 fluorescence transients after field stimulation were confined to the cytosol and nucleus. In contrast, after loading at 4 degrees C, Fluo 3 also entered mitochondria, and large transients of mitochondrial Fluo 3 fluorescence then occurred after stimulation. Isoproterenol (1 microM) increased the magnitude of Ca2+ transients and their subsequent rate of decay, an effect more marked in the cytosol and nucleus than in mitochondria. As pacing frequency was increased from 0.5 to 2 Hz, diastolic mitochondrial Ca2+ rose markedly in the absence but not in the presence of isoproterenol. Resting Ca2+ estimated by Indo 1 ratio imaging using UV/visible laser scanning confocal microscopy was about 200 nM in all compartments. During field stimulation, Ca2+ transiently increased to 671, 522, and 487 nM in cytosol, interfibrillar mitochondria, and perinuclear mitochondria, respectively. Isoproterenol increased these respective peak values to 1280, 750, and 573 nM. These results were consistent with those obtained in Fluo 3 experiments. We conclude that rapid mitochondrial Ca2+ transients occur during excitation contraction coupling in adult rabbit cardiac myocytes, which may be important in matching mitochondrial metabolism to myocardial ATP demand during changes in cardiac output. PMID- 9733090 TI - Localization at complex I and mechanism of the higher free radical production of brain nonsynaptic mitochondria in the short-lived rat than in the longevous pigeon. AB - Free radical production and leak of brain nonsynaptic mitochondria were higher with pyruvate/malate than with succinate in rats and pigeons. Rotenone, antimycin A, and myxothiazol maximally stimulated free radical production with pyruvate/malate but not with succinate. Simultaneous treatment with myxothiazol plus antimycin A did not decrease the stimulated rate of free radical production brought about independently by any of these two inhibitors with pyruvate/malate. Thenoyltrifluoroacetone did not increase free radical production with succinate. No free radical production was detected at Complex IV. Free radical production and leak with pyruvate/malate were higher in the rat (maximum longevity 4 years) than in the pigeon (maximum longevity 35 years). These differences between species disappeared in the presence of rotenone. The results localize the main free radical production site of nonsynaptic brain mitochondria at Complex I. They also suggest that the low free radical production of pigeon brain mitochondria is due to a low degree of reduction of Complex I in the steady state in this highly longevous species. PMID- 9733089 TI - Cloning and sequence analysis of the structural gene for the bc1-type Rieske iron sulfur protein from Thermus thermophilus HB8. AB - The structural gene encoding the Rieske iron-sulfur protein from Thermus thermophilus HB8 has been cloned and sequenced. The gene encodes a protein of 209 amino acids that begins with a hydrophilic N-terminus followed by a stretch of 21 hydrophobic amino acids that could serve as a transmembrane helix. The remainder of the protein has a hydrophobicity pattern typical of a water-soluble protein. A phylogenetic analysis of 26 Rieske proteins that are part of bc1 or b6f complexes shows that they fall into three major groups: eubacterial and mitochondrial, cyanobacterial and plastid, and five highly divergent outliers, including that of Thermus. Although the overall homology with other Rieske proteins is very low, the C-terminal half of the Thermus protein contains the signature sequence CTHLGC (13X)-CPCH that most likely provides the ligands of the [2Fe-2S] cluster. It is proposed that this region of the protein represents a small domain that folds independently and that the encoding DNA sequence may have been transferred during evolution to several unrelated genes to provide the cluster attachment site to proteins of different origin. The role of individual residues in this domain of the Thermus protein is discussed vis-a-vis the three-dimensional structure of the bovine protein (Iwata et al., 1996 Structure 4, 567-579). PMID- 9733091 TI - Binding of rat brain hexokinase to recombinant yeast mitochondria: effect of environmental factors and the source of porin. AB - Heterologous binding of rat brain hexokinase to wild type, porinless, and recombinant yeast mitochondria expressing human porin was assessed, partially characterized, and compared to that in the homologous system (rat liver mitochondria). With porin-containing yeast mitochondria it is shown that (i) a significant, saturable association occurs; (ii) its extent and apparent affinity, correlated with the origin of porin, are enhanced in the presence of dextran; (iii) the binding requires Mg ions and apparently follows a complex cooperative mechanism. This heterologous association does not seem to differ fundamentally from that in the homologous system and represents a good basis for molecular studies in yeast. With porinless yeast mitochondria, binding occurs at much lower affinity, but to many more sites per mitochondrion. The results indicating a major but not exclusive role for porin in the binding are discussed in terms of (i) the mode and mechanism of binding, and (ii) the suitability of the rat hexokinase-yeast mitochondria couple for the study of heterogeneous catalysis in reconstituted cellular model systems. PMID- 9733092 TI - Channel specificity and secondary structure of the glucose-inducible porins of Pseudomonas spp. AB - The OprB porin-mediated glucose transport system was investigated in Pseudomonas chlororaphis, Burkholderia cepacia, and Pseudomonas fluorescens. Kinetic studies of [U-14C]glucose uptake revealed an inducible system of low Km values (0.3-5 microM) and high specificity for glucose. OprB homologs were purified and reconstituted into proteoliposomes. The porin function and channel preference for glucose were demonstrated by liposome swelling assays. Examination of the periplasmic glucose-binding protein (GBP) components by Western immunoblotting using P. aeruginosa GBP-specific antiserum revealed some homology between P. aeruginosa GBP and periplasmic proteins from P. fluorescens and P. chlororaphis but not B. cepacia. Circular dichroism spectropolarimetry of purified OprB-like porins from the three species revealed beta sheet contents of 31-50% in agreement with 40% beta sheet content for the P. aeruginosa OprB porin. These findings suggest that the high-affinity glucose transport system is primarily specific for glucose and well conserved in the genus Pseudomonas although its outer membrane component may differ in channel architecture and specificity for other carbohydrates. PMID- 9733093 TI - Hypothyroidism leads to a decreased expression of mitochondrial F0F1-ATP synthase in rat liver. AB - In liver mitochondria isolated from hypothyroid rats, the rate of ATP synthesis is lower than in mitochondria from normal rats. Oligomycin-sensitive ATP hydrolase activity and passive proton permeability were significantly lower in submitochondrial particles from hypothyroid rats compared to those isolated from normal rats. In mitochondria from hypothyroid rats, the changes in catalytic activities of F0F1-ATP synthase are accompanied by a decrease in the amount of immunodetected beta-F1, F0 1-PVP, and OSCP subunits of the complex. Northern blot hybridization shows a decrease in the relative cytosolic content of mRNA for beta F1 subunit in liver of hypothyroid rats. Administration of 3,5,3'-triodo-L thyronine to the hypothyroid rats tends to remedy the functional and structural defects of F0F1-ATP synthase observed in the hypothyroid rats. The results obtained indicate that hypothyroidism leads to a decreased expression of F0F1-ATP synthase complex in liver mitochondria and this contributes to the decrease of the efficiency of oxidative phosphorylation. PMID- 9733096 TI - Genetic analysis of a conserved sequence in the HoxD complex: regulatory redundancy or limitations of the transgenic approach? AB - Extensive sequencing in the HoxD complex of several vertebrate species has revealed a set of conserved DNA sequences interspersed between neighboring Hox genes. Their high degree of conservation strongly suggested that they are used for regulatory purposes, a hypothesis that was largely confirmed by using "classical transgenesis" or in vivo mutagenesis through the embryonic stem (ES) cell technology. Here, we show that this is not always the case. We report that the deletion of a conserved regulatory sequence located in the HoxD complex gives different results, depending on the transgenic approach that was used. In "conventional" transgenesis, this sequence was necessary for proper expression in a subdomain of the developing limb. However, a deletion of this sequence in complexo did not confirm this effect, thereby creating an important discrepancy between the classical transgenic and the ES cell-based, targeted mutagenesis. This unexpected observation may show the limitations of the former technology. Alternatively, it could illustrate a redundancy in regulatory circuits and, thus, justify the combination of parallel strategies. PMID- 9733095 TI - Plant cell membranes as biochemical targets of the phytotoxin helminthosporol. AB - Helminthosporol is one of the natural sesquiterpenoid toxins isolated and identified in the culture medium of the phytopathogenic ascomycete fungus Cochliobolus sativus. The effect of this phytotoxin was investigated on enzymatic activities, electron and ion transport in mitochondria, chloroplasts, and microsomes of plant. The results indicate that helminthosporol drastically affects the membrane permeability of these organelles to protons and substrate anions, inhibiting the mitochondrial oxidative phosphorylation, the photophosphorylation in chloroplasts, and the proton pumping across the cell plasma membrane. The 1,3-beta-glucan synthase activity, involved in defense mechanisms of plant cells against stress and damage, e.g., during pathogen attack, was also strongly inhibited by the toxin. PMID- 9733094 TI - Human mitochondrial transmembrane metabolite carriers: tissue distribution and its implication for mitochondrial disorders. AB - Mitochondrial transmembrane carrier deficiencies are a recently discovered group of disorders, belonging to the so-called mitochondriocytopathies. We examined the human tissue distribution of carriers which are involved in the process of oxidative phosphorylation (adenine nucleotide translocator, phosphate carrier, and voltage-dependent anion channel) and some mitochondrial substrate carriers (2 oxoglutarate carrier, carnitine-acylcarnitine carrier, and citrate carrier). The tissue distribution on mRNA level of mitochondrial transport proteins appears to be roughly in correlation with the dependence of these tissues on mitochondrial energy production capacity. In general the main mRNA expression of carriers involved in mitochondrial energy metabolism occurs in skeletal muscle and heart. Expression in liver and pancreas differs between carriers. Expression in brain, placenta, lung, and kidney is lower than in the other tissues. Western and Northern blotting experiments show a comparable HVDAC1 protein and mRNA distribution for the tested tissues. Patient's studies showed that cultured skin fibroblasts may not be a reliable alternative for skeletal muscle in screening for human mitochondrial carrier defects. PMID- 9733097 TI - Temporal and spatial expression of alternative splice-forms of the alpha1(XI) collagen gene in fetal rat cartilage. AB - Type XI collagen, a member of the group of fibrillar collagens, plays a regulatory role in the formation of the collagen fibril network in cartilage and consequently plays a pivotal role in the formation of the endochondral skeleton. The mechanism by which type XI collagen limits fibril growth appears to involve the large noncollagenous amino terminal domain. Complex alternative splicing occurs within this domain in two of the three constituent subunits, alpha1(XI) and alpha2(XI). In the alpha1(XI) chain, three alternatively spliced exons encoding one very basic and two very acidic peptides generate six spliceforms and protein isoforms. In order to better understand the significance of this alternative splicing, we have examined fetal rat cartilage to determine: (a) the relationship between alternative splicing and chondrogenesis in limb bud micromass culture; (b) the relative levels of expression of each of the splice forms by ribonuclease protection; and (c) the distribution of splice-forms and protein isoforms by in situ hybridization and immunohistochemistry. The results indicate that the pattern of alternative splicing of the alpha1(XI) chain is tightly linked to chondrogenesis. The two most abundant spliceforms in fetal rib cartilage are v(o), lacking all three exons, and v1b, containing the exon encoding the basic peptide. While most of the spliceforms show a general distribution in nasal, Meckel's, and rib cartilage, v1b was restricted to the dorsal portion of the fetal rib. This distribution appears to correlate with the portion of the rib which will ultimately ossify, rather than with any of the differentiative states of chondrocytes. Together these results suggest that alternative splicing within the amino terminal domain of the alpha1(XI) chain may contribute to the function of type XI collagen and that expression of the basic v1b peptide may play a role in endochondral ossification. PMID- 9733098 TI - Expression and function of the urokinase type plasminogen activator during mouse hemochorial placental development. AB - Mouse midlate placental development involves extensive tissue remodeling and cell invasion, processes which could be mediated by extracellular proteolytic enzymes. We have performed in situ expression analysis of urokinase type plasminogen activator (uPA), as well as functionally related molecules (uPA receptor, low density lipoprotein receptor-related protein, plasminogen activator inhibitor type-1) in day 10.5 to 18.5 post coitum (p.c.) murine placentas. In situ hybridization demonstrated the presence of uPA transcripts in the invasive trophoblast cells, in particular in glycogen-rich trophoblasts, a cell population that between embryonic days 12.5 and 15.5 infiltrates the maternal decidual tissue. In addition, we observed high uPA expression in the cells of uterine epithelium. Enzymatically active uPA was detected in both sites of uPA mRNA expression by in situ zymography. Expression and activity data suggest a role for this protease in the processes of cell invasion and uterine epithelial remodeling. Only low levels of uPA receptor (uPAR) transcripts were found in trophoblasts and decidual tissue at days 10.5 and 11.5 p.c. At the same stages, a prominent expression of plasminogen activator inhibitor type-1 (PAI-1) by spongiotrophoblasts and giant trophoblasts, as well as of LDL receptor-related protein (LRP) by spongiotrophoblasts and decidual cells could be detected, suggesting a role in regulating extracellular proteolysis in the area of fetomaternal interface. Analysis of uPA null placentas showed the presence of decidual extravascular fibrin deposits, which were not detected in wild type placentas. At the same time, the extent of infiltration of trophoblast cells in maternal decidual tissue, evaluated by anti-cytokeratin immunostaining, was similar in wild type and uPA null placentas. Our studies show that in murine hemochorial placentation, uPA has an essential role in the maintenance of the fibrinogenic/fibrinolytic balance in the decidua. The function of uPA in trophoblast invasion appears not to be indispensable, and its absence can be overcome by redundant or compensatory mechanisms. PMID- 9733099 TI - Graded retinoid responses in the developing hindbrain. AB - The purpose of this study was to make an explicit test of the idea that a retinoid could act as a morphogen, differentially activating genes and specifying anteroposterior (a-p) level in the developing vertebrate central nervous system (CNS). Our approach was to characterize the concentration-dependent effects of retinoic acid (RA) on the neural expression of a set of a-p patterning genes, both in vivo and in an in vitro system for neural patterning. Our results indicate that a retinoid is unlikely to specify a-p level along the entire CNS. Instead, our data support the idea that the developing hindbrain may be patterned by a retinoid gradient. Sequentially more posterior hindbrain patterning genes were induced effectively by sequentially higher RA concentration windows. The most posterior CNS level induced under our RA treatment conditions corresponded to the most posterior part of the hindbrain. PMID- 9733100 TI - Diminished growth of atrioventricular cushion tissue in stage 24 retinoic acid treated chicken embryos. AB - Stage 34 chicken hearts have shown a spectrum of looping disturbances, changed hemodynamics, and changed growth of both right ventricular myocardium and atrioventricular cushion tissue after retinoic acid treatment. To obtain more information about the onset of the malformations we studied stage 24, the stage between the previously studied stage 34 and the moment of treatment. Sixteen stage 24 chicken embryos were examined after treatment with 1 microg all-trans retinoic acid at stage 15 and compared with 6 sham operated embryos. Morphological examination was supported by graphic reconstructions. Absolute volumes of atrial, atrioventricular, and ventricular myocardia were measured by a point counting method. The absolute volumes of the endocardial cushions were measured as well. Fifteen (15/16) retinoic acid-treated hearts did not show marked malformations as far as could be detected with our current macroscopic and microscopic techniques. One (1/16) retinoic acid-treated heart showed an abnormal tubular C-shape with a less bended inner curvature and with an abnormal horizontally oriented atrioventricular canal. The dorsal cushion tissue of this atrioventricular canal was discontinuous with the dorsal mesocardium and covered the malpositioned myocardial border between the atrium and the atrioventricular canal. The volume measurements did show a difference between retinoic acid treatment and sham operations. The retinoic acid-treated hearts showed a significant volume decrease of the atrioventricular cushions. No significant differences were found in the volumes of the ventricular myocardium compared to the sham operated embryos. We hypothesize that, between stages 15 and 24, retinoic acid directly affects the myocardial wall and the cushion tissue formation. In the present material this has resulted in decreased atrioventricular cushion growth, in changed hemodynamics, and in a severe looping disturbance of one embryo. We further hypothesize that, between stages 24 and 34, the malformations with minor looping disturbances will become apparent. Thus, development beyond stage 24 would result in the spectrum of looping disturbances as has been found at stage 34. These latter morphological malformations would lead to increasing hemodynamic changes, resulting in changes in growth as a secondary effect. PMID- 9733101 TI - Patterns of paired-related homeobox genes PRX1 and PRX2 suggest involvement in matrix modulation in the developing chick vascular system. AB - PRX1 (MHox) and PRX2 (S8) were previously shown to be expressed throughout embryogenesis in complex, mostly mesenchyme-specific patterns. In the developing cardiovascular system both genes were highly expressed in prospective connective tissues, that is, endocardial cushions and valves, the epicardium, and the wall of the great arteries and veins. We further scrutinised expression of PRX1 and PRX2 in the developing vascular system of the chicken embryo and compared patterns with those of established vascular differentiation markers (muscle actin, procollagen I, and fibrillin-2). PRX1 and PRX2 expression were associated with the primary vessel wall from early stages onward and became increasingly restricted to the adventitial and outer medial cell layers. PRX1 eventually colocalised strikingly with procollagen I and fibrillin-2 expression and generally excluded high smooth muscle actin expression. Furthermore, PRX1 expression preceded the segregation of very distinct nonmuscular cells and smooth muscle cells in the media of the great arteries. PRX2 patterns deviated at later stages from those of PRX1 and showed specific and high transcript levels in the ductus arteriosus from embryonic day 6 onward. Results suggest that PRX genes are not essential in smooth muscle contractile differentiation, but may be involved in matrix modulation in the vascular system and possibly in defining the noncontractile cellular phenotype and in media-adventitia definition. PMID- 9733102 TI - Transforming growth factor alpha up-regulates desmin expression during embryonic mouse tongue myogenesis. AB - Myogenesis is determined by a set of myogenic differentiation factors that are, in turn, regulated by a number of peptide growth factors. During embryonic mouse tongue formation, transforming growth factor alpha (TGF alpha), epidermal growth factor (EGF), and their cognate receptor (EGFR) are co-expressed spatially and temporally with desmin, a muscle-specific structural protein. This investigation tested the hypothesis that TGF alpha directly regulates the myogenic program in developing tongue myoblasts. Mandibular processes from the first branchial arch of embryonic day 10.5 (E10.5) mouse embryos were microdissected and explanted into an organ culture system using serumless chemically defined medium. Exogenous TGF alpha at 10 and 20 ng/ml specifically increased the amount of desmin expression and the number of desmin-positive cells without affecting the general growth and development of the mandibles. This inductive response was detected as early as 2 days after treatment and sustained up to 9 days in culture. EGFR antisense oligonucleotides (30 microM) as well as tyrphostin (80 microM) were able to negate TGF alpha-induced up-regulation of desmin expression. These data indicate that autocrine and/or paracrine action of TGF alpha promotes tongue myogenesis, and that this action is mediated through functional kinase activity of the EGFR. We speculate that the myogenic program in the developing mouse tongue is dependent upon growth factor mediated cell-cell communication of mesenchymal cells originating from the occipital somites and ectomesenchymal cells originating from the cranial neural crest. PMID- 9733103 TI - Analysis of Hox gene expression during early avian heart development. AB - The anteroposterior (A-P) patterning of the developing heart underlies atrial and ventricular lineage specification and heart chamber morphogenesis. The posteriorization of cardiomyogenic phenotype with retinoic acid (RA) treatment of primitive streak stage chicken embryos is suggestive of a role for the clustered homeobox (Hox) genes in early heart patterning (Yutzey et al. [1994] Development 120:871-873; [1995] Dev. Biol. 170:531-541). A screen for Hox genes expressed in chick heart primordia and primitive heart led to the isolation of anterior genes of the Hox clusters expressed during cardiogenesis. Specific hoxd-3, hoxa-4, and hoxd-4 transcripts were detected at the early stages of heart formation and full length cDNA clones were isolated. Expression of hoxd-3 was detected in the heart forming region of embryos prior to heart tube formation. Expression of hoxa-4, hoxd-3, and hoxb-5 was increased in cardiogenic tissue treated with RA in culture conditions that also produced changes in positionally restricted cardiomyogenic phenotypes. Hox genes expressed in cardiac explants exhibited distinct sensitivities to RA and ouabain treatment when compared to genes, such as nkx 2.5, that are involved in cardiac commitment and differentiation. These studies support a role for Hox genes in early heart patterning and suggest that positional information in the cardiogenic region is established by regulatory mechanisms distinct from early heart lineage specification. PMID- 9733104 TI - Expression of zebrafish bHLH genes ngn1 and nrd defines distinct stages of neural differentiation. AB - Two zebrafish bHLH genes, neurogenin-related gene I (ngn1) and neuroD (nrd), have been isolated. ngn1 expression is initiated at the end of gastrulation in the neural plate and defines broad domains of cells that probably possess an ability to develop as neurons. This finding suggests that ngn1 may play a role during determination of cell fate in neuroblasts. ngn1 and pax-b are expressed in a mutually exclusive manner. nrd expression follows that of ngn1 in restricted populations of cells selected from ngn1-positive clusters of cells. The earliest nrd-positive cells in the brain and the trunk are a subset of the primary neurons. ngn1 is not expressed in the eye. Here, nrd transcription is activated at 25 hours postfertilization in the ventral retina. Expression of islet-1 occurs in nrd-positive cells after expression of nrd, and the expression of the two genes partially overlaps in time. These observations suggest that during eye development nrd expression may follow expression of some other neurodetermination gene(s). This supports the idea that expression of nrd is a necessary step leading toward overt neuronal differentiation. PMID- 9733105 TI - epicardin: A novel basic helix-loop-helix transcription factor gene expressed in epicardium, branchial arch myoblasts, and mesenchyme of developing lung, gut, kidney, and gonads. AB - We report the cloning, chromosomal localization, and analysis of the expression pattern of epicardin, a member of the basic helix-loop-helix (bHLH) family of transcription factors. Within its bHLH domain, the human and murine epicardin genes were most similar to paraxis, a bHLH gene important for segmentation of embryonic paraxial mesoderm. In situ hybridization studies revealed strong epicardin expression in murine embryos at 9.5 days postcoitum (dpc) in a region of the septum transversum at the base of the heart known as the proepicardial organ. This mesenchymal structure extends villous projections from which epicardial precursor cells emerge and migrate out over the surface of the myocardium. Strong expression was seen in individual migratory cells and clusters at 9.5 dpc and in a continuous epicardial cell layer in more mature hearts. Also from 9.5 dpc, epicardin transcripts were seen in endocardial cushions of the atrioventricular canal and outflow tract, in skeletal myoblasts within branchial arches and in condensing mesenchyme of gut, kidney, urinary tract, gonads, spleen, and lung. Northern analysis showed that expression persisted in mature visceral organs and heart, but was transient in skeletal muscle. The central role played by bHLH factors in pathways for tissue determination in the embryo suggests a function for epicardin in specification of select mesodermal cell populations associated with heart, cranial skeletal muscle, gut, and urogenital system. PMID- 9733106 TI - ADH1 and ADH4 alcohol/retinol dehydrogenases in the developing adrenal blastema provide evidence for embryonic retinoid endocrine function. AB - Studies on retinoid signaling indicate that much of the regulation of this pathway may involve enzymes that synthesize the active ligand retinoic acid. Alcohol dehydrogenases ADH1 (class I ADH) and ADH4 (class IV ADH) function as retinol dehydrogenases in the oxidation of retinol, a necessary step in the synthesis of retinoic acid from vitamin A. These enzymes as well as retinoic acid have previously been localized in the adult adrenal gland, thus providing evidence that this organ is an endocrine source of retinoic acid. Here, we have examined the involvement of ADH1 and ADH4 in embryonic adrenal function by using transgenic mouse technology and immunohistochemistry. Transgenic mice were generated that contain various portions of the mouse ADH4 promoter and 5' flanking region fused to lacZ. Embryos harboring a construct containing 9.0 kb of 5'-flanking region displayed very high levels of lacZ expression in the developing adrenal blastemas at embryonic stage E11.5 during the initial phase of mouse adrenal gland development. The presence of endogenous ADH4 protein in stage E11.5 adrenal blastemas was demonstrated by immunohistochemistry, and this was the only site of ADH4 immunodetection in stage E11.5 embryos. Endogenous ADH1 protein was also detected by immunohistochemistry in stage E11.5 adrenal blastemas. ADH1 and ADH4 proteins were detectable at later stages of adrenal development, and both were localized to developing adrenal cortical cells by stage E14.5. The presence of both ADH1 and ADH4 retinol dehydrogenases during the earliest stages of adrenal gland development, combined with our earlier findings of high levels of retinoic acid in the embryonic adrenal gland, suggests that one of the earliest functions of ADH may be to provide an embryonic endocrine source of retinoic acid for growth and development. PMID- 9733107 TI - Regulated expression of matrix metalloproteinases and TIMP in nephrogenesis. AB - The roles of the matrix metalloproteinases (MMPs) and their specific inhibitors, the tissue inhibitors of metalloproteinases (TIMP), in embryologic development in general, and in nephrogenesis in particular, have not been fully elucidated. The activities of these enzymes and their inhibitors may be critical in the extensive extracellular matrix remodeling that accompanies the formation of the full complement of mature nephrons in the developing kidney. The temporal and spatial expression of two critical basal lamina modifying enzymes, the 72 kDa gelatinase A (MMP-2) and the 92 kDa gelatinase B (MMP-9), as well as TIMP-1, -2, and -3 molecules were evaluated in the developing rat kidney. Additionally, transcripts for the recently described membrane-associated matrix metalloproteinase, MT1-MMP (MMP-14), which can act as an activating receptor for MMP-2/TIMP-2 complexes (Strongin et al.[1995] J. Biol. Chem. 270:5331-5338) were localized by in situ hybridization. Our immunohistochemical data demonstrate distinct localization of MMP-2 within immature nephron structures undergoing epithelial differentiation, while MMP-9 localizes only to the invading vascular structures within immature glomeruli. In contrast, by in situ hybridization, MMP-2 transcripts localize to the background undifferentiated mesenchyme and not to those structures undergoing epithelial differentiation. In a pattern similar to the MMP-2 protein, MT1-MMP transcripts were found within developing epithelial structures. Neither MMP-2, MMP-9 nor MT1-MMP were detected in mature nephrons. TIMP-2 and -3 follow a pattern of expression similar to the MMP-2 protein. We conclude that MMP-2 and TIMP play important roles in the remodeling of basal laminae associated with the epithelial structures of the developing kidney, that these enzymes are temporally and spatially regulated, and that the co-localization of MT1-MMP to sites of basement membrane remodeling suggests a potential role for this molecule as a receptor for and/or modulator of MMP-2/TIMP complexes. PMID- 9733108 TI - AmphiBMP2/4, an amphioxus bone morphogenetic protein closely related to Drosophila decapentaplegic and vertebrate BMP2 and BMP4: insights into evolution of dorsoventral axis specification. AB - Amphioxus AmphiBMP2/4 appears to be a single gene closely related to vertebrate BMP2 and BMP4. In amphioxus embryos, the expression patterns of AmphiBMP2/4 suggest patterning roles in the ectodermal dorsoventral axis (comparable to dorsoventral axis establishment in the ectoderm by Drosophila decapentaplegic and vertebrate BMP4). In addition AmphiBMP2/4 may be involved in somite evagination, tail bud growth, pharyngeal differentiation (resulting in club-shaped gland morphogenesis), hindgut regionalization, differentiation of olfactory epithelium, patterning of the anterior central nervous system, and establishment of the heart primordium. One difference between the developmental role of amphioxus AmphiBMP2/4 and vertebrate BMP4 is that the former does not appear to be involved in the initial establishment of the dorsoventral polarity of the mesoderm. PMID- 9733109 TI - Regulation of lens regeneration by fibroblast growth factor receptor 1. AB - Lens regeneration in vivo is restricted to some urodeles only. After removal of the lens, this remarkable event is initiated from the dorsal iris. The pigmented epithelial cells from the dorsal iris dedifferentiate and subsequently transdifferentiate to form the regenerating lens. This property of the dorsal iris implies specific regulation along the dorsal-ventral axis. To date, no known genes are known to be specifically expressed in the dedifferentiating cells and to be involved in lens regeneration. In this paper, we show that FGFR-1 expression and function is correlated with the process of lens regeneration from the dorsal iris. Following lentectomy, FGFR-1 protein is specifically present in the dedifferentiating pigment epithelial cells in the dorsal iris, but is absent from the ventral iris. Subsequently, FGFR-1 protein is present throughout the process of lens regeneration and fiber differentiation. Furthermore, we show that an FGFR-1-specific inhibitor is able to inhibit the process of transdifferentiation and lens regeneration. In this sense, FGFR-1 can be regarded as the first known lens regeneration-associated factor. PMID- 9733110 TI - Primary myotubes preferentially mature into either the fastest or slowest muscle fibers. AB - Myoblasts and myotubes are heterogeneous, but what is the significance of this heterogeneity? Is it a vital component of the mechanism by which a muscle develops or is it part of the process that generates mature fibers with diverse sizes, speeds of contracture, and metabolisms? We have begun to explore these questions by using BrdU to selectively label rat primary myotubes, thus enabling their mature characteristics to be defined for the first time. In the soleus, the type I fibers of primary myotube origin were 21% larger than those of secondary myotube origin, indicating that the origin of a fiber can affect its mature force production. In the extensor digitorum longus (EDL), the primary myotubes differentiated into all known fibers types, but with marked variation in frequency. In the superficial portion of the EDL, 97% of primary myotubes became IIB fibers, even though approximately 41% of the fibers in this region are IIA or IIX. In the deep portion, primary myotubes preferentially developed into type I fibers. Thus, primary myotubes in the EDL predominantly differentiate into the two most dissimilar fiber types: the slowest, smallest, most oxidative, type I fibers and the largest, fastest, most glycolytic, type IIB fibers. Each of the subtypes of primary myotubes had a different fate. In the EDL, the slow and fast primary myotubes appeared to differentiate into type I and IIB fibers, respectively. This implies that spatial and temporal signals operating in the limb are major determinants of the mature pattern of fiber types and that innervation of a muscle involves a selective matching between the various types of motoneurons and muscle fibers. PMID- 9733111 TI - Growth performance, intestinal microbial populations, and serum cholesterol of broilers fed diets containing Lactobacillus cultures. AB - A study was conducted to determine the effects of adherent Lactobacillus culture on growth performance, intestinal microbial population, and serum cholesterol level of broilers. Four dietary treatments, consisting of the basal diet (control), basal diet + 0.05, 0.10, or 0.15% Lactobacillus culture (LC), were fed to 2,000 Arbor Acres broiler chicks from 1 to 42 d of age (DOA). The chicks were randomly assigned to 40 cages (50 chicks per cage, 10 cages per diet). The experimental period was 42 d. Body weights and feed to gain ratio were measured at 21 and 42 DOA. The intestinal microbial populations and serum cholesterol levels were determined at 10, 20, 30, and 40 DOA. The results showed that body weights and feed to gain ratios were improved significantly (P < 0.05) when compared to control broilers for broilers fed diets containing 0.05 or 0.10% LC, but not 0.15% LC, at 21 and 42 DOA. Coliform counts in the cecum of birds receiving 0.05% LC at 10, 20, and 30 DOA, and 0.10% at 10 and 20 DOA were significantly lower (P < 0.05) than those of the control birds. The total aerobes, total anaerobes, lactobacilli, and streptococci in the small intestines and ceca of the control birds were not significantly different from those of the treated groups. Serum cholesterol levels were significantly lower (P < 0.05) in broilers fed the three diets containing LC at 30 DOA, and in the birds fed 0.05 or 0.10% LC at 20 DOA. PMID- 9733112 TI - The effect of light regimen, floor space, and energy and protein levels during the growing period on body weight and early egg size. AB - Two experiments (Exp.) were conducted to determine whether dietary manipulation of energy and protein, additional floor space, and the use of a step-down light regimen during the growing period can influence BW of the White Leghorn pullets at the age of housing (18 wk) and egg size during the early stages of egg production. The results of Exp. 1 indicated that BW at 18 wk of age was increased (P < 0.05) due to the use of high energy (3,036 vs 2,816 kcal ME/kg) or high protein (17.5 vs 14.5%) diets or providing the pullets with more floor space (346 vs 283 cm2 per pullet) from 8 to 18 wk of age. However, the extent of increases of BW due to these variables were not large enough to increase egg size during the early stages of the egg production cycle. The results of Exp. 2 indicated that 18-wk BW was increased (P < 0.05) only due to the use of a high protein sequence (22, 18, and 16% vs 18, 16, and 14% that were used during 0 to 6, 6 to 12, and 12 to 18 wk of age, respectively). Body weight at 18 wk of age was not influenced by the use of a higher energy diet (3,036 vs 2,816 kcal ME/kg) from day-old to 18 wk. Energy or protein levels did not have an effect on early egg size or overall performance up to 66 wk of age. Body weight of the pullets on the step-down light regimen (which were exposed to 23 h/d light at day-old and was gradually reduced to 8 h/d at 15 wk of age) were heavier than the pullets of the short-day light regimen (which were exposed to 8 h/d light during the growing period) during most parts of the growing and laying periods (P < 0.05). The step down light regimen resulted in increased egg size and higher percentage of extra large plus large-sized eggs for the entire experiment (P < 0.05), but reduced hen day egg production and egg mass and impaired feed conversion (P < 0.05) for the entire experiment (18 to 66 wk of age). PMID- 9733113 TI - Breeding biology of Muscovy duck Cairina moschata in natural incubation: the effect of nesting behavior on hatchability. AB - A breeding biology study of a genetically unselected variety of Muscovy duck Cairina moschata was conducted in an experimental duck unit in Maputo, Mozambique (25 degrees 58' S, 32 degrees 35' E), to gain insight into factors affecting hatchability during natural incubation. Nesting and incubation behaviors were recorded by daily nest visits. Of 1,338 nests investigated, 70% were incubated until hatching. In 23% of the incubated nests, eggs were laid by more than one duck in the same nest, indicated as dump nests. Artificial dump nests (15% of the incubated nests) were created by adding eggs (690 eggs) from nests abandoned by the duck before incubation had started. A total of 37% of the incubated nests contained eggs that were laid after the onset of incubation (nonterm eggs). Similar hatching rate was found between ducks raised in parks with or without access to swimming water. No significant difference was found in hatchability between normal (0.76) and dump nests (0.77). Artificial dump nests showed higher hatching rates than nests containing nonterm eggs. Dump nesting appears to be a reproductive strategy used by the Muscovy duck to enhance duckling production. Hatching rate was strongly influenced by the length of laying period (period between the ovoposition of the first egg until the onset of incubation) and reproduction cycle (laying period and incubation period). Hatchability was higher for clutches with a shorter reproduction cycle. It is concluded that nesting behavior of the domesticated Muscovy duck is similar to that of its wild ancestor. PMID- 9733114 TI - Comparative effects of added sodium chloride, ammonium chloride, or potassium bicarbonate in the drinking water of broilers, and feed restriction, on the development of the ascites syndrome. AB - A hypothesis that the ionic composition of drinking water might affect development of the ascites syndrome in broilers was investigated in two trials. The first trial comprised four groups of 650 male chicks. A control treatment was normal tap water and the other three treatments comprised the addition to the tap water of 1,000 mg/L sodium as NaCl, 5,000 mg/L NH4Cl, or 5,000 mg/L KHCO3, supplied from age 2 to 47 d. At Day 28, equally sized subsets of these groups were moved to individual cages, where they received a severe exposure to ambient cold. The development of the ascites syndrome was monitored by measurements of hematocrit and arterial blood oxygen saturation (PaO2) by oximetry, body weight, and examination of dead birds for cause of death. Mortality from ascites in cold exposed birds from Days 28 to 47 was 28, 48, 40, and 16% in the tap water, NaCl, NH4Cl, and KHCO3 groups, respectively; only the NaCl mortality was significantly different from the tap water mortality. The KHCO3 treatment increased PaO2 (compared with tap water treatment) at Day 28 by 5.5% and at Day 35 by 10.5%, but not at Day 42. The KHCO3 caused a reduction in body weight, which was 13% less than the tap water group at Day 42, probably due to a chronic toxicity. The second trial specifically examined the same parameters with lower water levels of KHCO3 (3,000 and 1,000 mg/L), in comparison to a 10% feed restriction protocol, in order to clarify whether the increased PaO2 was due to a specific effect of the KHCO3 or was a metabolic manifestation of a reduced growth rate. The 3,000 mg/L KHCO3 treatment had no effect on PaO2, but the 1,000 mg/L treatment augmented PaO2 by 5.3% at Day 35 (but not at Days 28 or 42), without reducing the final body weight. The feed restriction group showed an elevated PaO2 of 5.4% at Day 35 (but not at Days 28 or 42), with no reduction in the final body weight. The inclusion of 1,000 mg/L of KHCO3 into the drinking water of broilers or a temporary 10% feed restriction may be means to augment PaO2. PMID- 9733115 TI - Use of low doses of clenbuterol to reduce incidence of ascites syndrome in broilers. AB - Beta-adrenergic agonists have been shown to be capable of improving growth performance in poultry when added to the feed at 1.0 ppm. However, no reference has been made concerning the cardiovascular responses when one of these agents is added to the feed at a lower concentration during the whole production cycle. The aim of this paper was to assess the effects on the ascites syndrome of 0.25 ppm clenbuterol in the feed, throughout 52 d, in broiler chicks. Results showed a lack of difference in growth and feed conversion rate between the untreated control groups and the experimental group. There were differences in mortality due to the ascites syndrome, abdominal fat:body weight ratio, and ventricular index. A statistically significant positive correlation was also found between ventricular index and mortality rate (r = 0.98). If adequate withdrawal times are ensured, the use of clenbuterol at 0.25 ppm is suggested to reduce mortality due to the ascites syndrome in broilers. PMID- 9733116 TI - Relationship between long bone distortion and tibial dyschondroplasia in male turkeys. AB - To determine whether a relationship exists between long bone distortion and tibial dyschondroplasia (TD), the hock joints of Large White commercial male turkeys were examined using low intensity x-ray imaging (hand-held lixiscope). All lame birds with long bone distortion and 96 control birds without lameness were examined at 8.3 and 12.3 wk of age for TD lesions using a lixiscope. Results indicated that the incidence of TD was less than 2% at 8.3 wk of age. The incidence increased to 21% by 12.3 wk of age. The TD lesions at 12 wk of age were considered mild covering only one-fourth of the growth plate. Chi-square analysis indicated that control turkeys without long bone distortion were just as likely to have TD lesions as were lame turkeys with long bone distortion. Under the conditions of the present experiment, TD did not appear to be a significant skeletal problem in Large White commercial male turkeys prior to 12.3 wk of age. Only 5 out of 37 (14%) toms with long bone distortion at 12.3 wk of age had TD lesions, suggesting that the valgus-varus deformity may be independent of TD. PMID- 9733117 TI - Incidence of Campylobacter in crops of preharvest market-age broiler chickens. AB - Previous research has identified cecal and intestinal contents as sources for Campylobacter contamination of broiler carcasses in the processing plant. During the present study, we evaluated the crop contents of preharvest market-age broilers as a potential reservoir of field-derived Campylobacter in the processing plant. Crops were collected aseptically from 40 randomly selected market-age broilers in each of nine commercial broiler flocks. Ceca were collected from broilers in six of the same flocks for comparison with the crop samples. The presence of Campylobacter in the crops and ceca was determined by enrichment culture in Bolton broth followed by culture on Campy-Ceflex plates. Campylobacter was isolated from the crop contents of broilers in seven of the nine flocks and from the cecal contents in three of six flocks. The incidence of Campylobacter-positive crop samples among all birds evaluated (224/359; 62%) was significantly higher (P < 0.001) than the number of positive cecal samples (9/240; 4%). The results indicate that the incidence of Campylobacter contamination of crop contents may exceed that of the cecal contents by as much as 37-fold in some broiler flocks, and may represent a critical preprocessing control point in reducing Campylobacter entry into the processing plant. PMID- 9733118 TI - Effect of tylosin tartrate (Tylan Soluble) on cellular immune responses in chickens. AB - Although many antimicrobial agents have been reported to cause immunosuppression in animals, macrolide antibiotics enhance immune function. Tylosin is a macrolide antibiotic approved for the control of mycoplasmosis in poultry. The purpose of this investigation was to determine the effect of tylosin on cellular immune functions in chickens. There was no significant difference in adherent splenocyte chemotaxis between tylosin-treated and untreated (control) chickens. Tylosin increased splenocyte proliferation and splenocyte conditioned medium (CM) proliferative activity above control levels. Removal of adherent splenocytes before preparation of CM caused a reduction in CM proliferative activity. Tylosin also increased antitumor activity of splenocytes. These data are the first to suggest that the macrolide antibiotic, tylosin tartrate, has a modulatory effect in chickens on the immune parameters studied. PMID- 9733119 TI - Bioavailability of tryptophan in soybean meal and tryptophan retention in the carcasses of four-week-old ducks. AB - The objectives of the experiment were to determine the bioavailability of tryptophan in soybean meal for 4-wk-old ducks using the slope-ratio bioassay with L-tryptophan as the reference diet and estimate the efficiency of dietary tryptophan retention in the carcasses of 4- to 7-wk-old ducks. An initial representative group of 12 ducks with an average body weight of 1,951 g (SD = 69) were killed for carcass compositional analysis. A basal diet formulated to contain 1.13 g of tryptophan/kg and adequate in all other amino acids, was supplemented with 0.2, 0.4, or 0.6 g tryptophan/kg from L-tryptophan, or 0.2 or 0.4 g tryptophan/kg from soybean meal. Four pens of three 4-wk-old ducks received each of the six diets for 21 d. Ducks fed the basal diet and the basal diet supplemented with 0.2 or 0.4 g of L-tryptophan/kg for 21 d were used as the final slaughter groups for carcass compositional analysis. Dietary supplemental tryptophan from either L-tryptophan or soybean meal linearly increased final weight (P < 0.05), weight gain (P < 0.10), and gain:feed ratio (P < 0.05). Common intercept, multiple linear regressions in slope-ratio methodology performed using weight gain and gain:feed ratio as dependent variables and grams of supplemental tryptophan/kg of diet as the independent variable gave 94 and 92% tryptophan bioavailability in soybean meal relative to L-tryptophan, respectively. Linear regression of tryptophan deposition in the carcass on tryptophan intake resulted in a 21% efficiency of carcass tryptophan retention above maintenance. PMID- 9733120 TI - Investigation on the possibility of reducing protein, phosphorus, and calcium requirements of laying hens by manipulation of time of access to these nutrients. AB - Experiments (Exp.) were conducted to determine whether the daily requirement of laying hens for protein, P, and Ca can be reduced by providing the hens with adequate levels of these nutrients only during those hours of the day that the physiological need for these nutrients for formation of various components of eggs are increasing. In Exp. 1, birds of the positive control were fed a 16% protein diet and birds of the negative control were fed a 13% protein diet continuously during the light period (0500 to 2100 h). The other groups were fed the 16% protein diet during the morning (0500 to 1300 h) and the 13% protein diet during the afternoon (1300 to 2100 h) or vice versa. The design of Exp. 2 was similar to Exp. 1. The birds of the positive control were fed a 0.4% available P (AP) and the birds of the negative control were fed a 0.2% AP diet, with other groups receiving the high-low AP or the low-high AP diets during the morning and the afternoon, respectively. The birds of the negative controls in these Exp. had almost comparable performance to those fed the other dietary treatments. As a result, these Exp. did not have negative controls for comparison of different dietary treatments. Additionally, regardless of dietary treatments, birds consumed about 40% of their daily feed intake during the morning and about 60% during the afternoon in these Exp. Due to these two shortcomings, it was not possible to reach to a decisive conclusion pertaining to the objectives of the Exp. The results of Exp. 3 indicated that the above pattern of daily feed intake was not due to an increased appetite for Ca during the afternoon hours for shell formation. Various indices of shell quality were not improved when most parts of the daily Ca need was fed during the afternoon and evening and were not reduced when most parts of the daily Ca need was fed during the morning. PMID- 9733121 TI - Further investigations on the effect of dietary manipulation of protein, phosphorus, and calcium for reducing their daily requirement for laying hens. AB - Experiments were conducted to determine the possibility of reducing the daily requirement of laying hens for Ca, available P (AP), and protein by providing the hens with adequate levels of these nutrients only during those hours of the day that the physiological need for these nutrients for formation of various components of the eggs are increasing. The results of the Ca experiment indicated that the daily Ca requirement cannot be reduced by providing the hens with adequate levels of Ca during the afternoon (1500 to 2100 h) and inadequate Ca level during the morning (0500 to 1500 h). Providing the hens with most of the daily Ca need during the afternoon did not have a beneficial effect on shell quality as compared to the control group that received a diet with 3.5% Ca during the morning and the afternoon. On the other hand, inadequate Ca intake during the afternoon adversely affected shell quality as compared to the control group (P < 0.05). The AP experiment indicated that egg production performance and shell quality can be maintained satisfactorily as long as the daily intake of AP is adequate, regardless of whether most of the AP is consumed during the morning or the afternoon. Egg production and shell quality of hens fed a diet with 0.4% AP during the morning and 0.1% AP during the afternoon or 0.1% AP during the morning and 0.4% AP during the afternoon were not different than the control group that received a diet with 0.25% AP during the morning and the afternoon. The results of two protein experiments failed to indicate that egg production performance of laying hens can be maintained satisfactorily by providing them with an adequate level of protein (16%) only during the morning and inadequate level of protein (10%) during the afternoon or, conversely, compared to the control group that received a diet with 16% protein both during the morning and afternoon (P < 0.05). The supply of adequate protein with sufficient amino acids, during the morning and the afternoon, was required for satisfactory maintaining egg production performance. Under the conditions selected for the conduct of this study, the results of the current experiments, combined with those of a previous report, failed to support the hypothesis that satisfactory performance may be maintained by providing the hens with adequate levels of protein, AP, and Ca only during those hours of the day when the physiological need for these nutrients for formation of various components of the egg are increasing. PMID- 9733122 TI - Developmental study of alpha-methyl-D-glucoside and L-proline uptake in the small intestine of the White Leghorn chicken. AB - Development changes in alpha-methyl-D-glucoside and L-proline accumulation were studied using everted sleeves from the duodenum, jejunum, and ileum of chickens. Six age groups of chickens were used: 1 d and 1, 2, 3, 5 to 6, and 12 to 14 wk. Our results showed the presence of a Na+-dependent mechanism of sugar and amino acid uptake that is already fully developed at hatch. The intestinal transport activity undergoes substantial changes with age and region studied. Intracellular accumulation of alpha-methyl-D-glucoside and L-proline was greater in newly hatched chicks and then declined with age in the three regions of the small intestine, except for L-proline transport in ileum, which remained constant during the period studied. The transport mechanisms for each nutrient followed separate developmental patterns along the small intestine during the period studied. PMID- 9733123 TI - Performance and tissue zinc and metallothionein accumulation in chicks fed a high dietary level of zinc. AB - Four experiments were conducted to identify several factors that might improve the accuracy and reproducibility of Zn bioavailability assays for chicks. Response of tissue Zn and metallothionein (MT) concentrations to various elevated levels and soluble sources of dietary Zn were measured, as well as the effect of delaying high Zn administration until 7 d posthatching to alleviate the detrimental effect of Zn sulfate on feed intake to 3 wk of age. Bone Zn increased (P < 0.01) in all experiments in response to increasing dietary Zn concentrations. Liver and pancreas MT were affected (P < 0.01) by a source by age interaction and variability that made this criterion unsuitable for bioavailability assays. Lastly, 1-d-old chicks were used to study the effect of delaying feeding of a high-Zn diet up to 7 d of age. The basal diet was fed continuously for 21 d as a control. A diet containing 1,000 ppm Zn was either fed continuously from Day 1, or started on Day 3, 5, or 7. Chicks given high Zn on Day 3, 5, or 7 decreased (P < 0.01) feed intake within 24 h of feeding. Delayed feeding of high dietary Zn might help to alleviate decreased feed intake observed in previous studies. Delaying the onset of high Zn feeding by several days may help alleviate feed intake problems observed with Zn sulfate. Use of either Zn gluconate or Zn acetate as a standard in assays or use of MT synthesis as a bioavailability criterion will probably not be useful to improve accuracy of the estimates. PMID- 9733124 TI - Dietary arginine and lysine in Large White toms. 1. Increasing arginine:lysine ratios does not improve performance when lysine levels are adequate. AB - A study was conducted utilizing two strains of male Large White turkeys (BUT Big 6 and Nicholas 700) to determine the effects of increasing Arg:Lys ratios on live performance and carcass composition. Diets were formulated to provide 100, 110, and 120% of NRC (1994) Lys levels, adjusted for dietary energy level, with Arg:Lys ratios of 1.0:1, 1.1:1, 1.2:1, and 1.3:1 in a 3 x 4 factorial arrangement. Eight pens of 15 poults (four pens of each strain) were fed each of the 12 test diets for an 18-wk period. Diets were changed at 3-wk intervals rather than the 4-wk interval suggested by NRC. The results of this study suggest that the Arg and Lys levels suggested by the NRC (1994) are not sufficient when diets are fed on 3-wk intervals, rather than the 4-wk intervals suggested by NRC. This conclusion is in agreement with the studies of Waldroup et al. (1997b). Increasing Arg:Lys ratios improved performance of turkeys only when the diets contained insufficient amounts of Arg in association with low levels of Lys. Increasing Arg:Lys ratios when diets contained sufficient amounts of these two amino acids was without benefit, in contrast to the report of Brake et al. (1994). Turkeys of the BUT Big 6 strain appeared to be more sensitive to marginal deficiencies of Lys and Arg than did turkeys of Nicholas 700 strain. PMID- 9733125 TI - Nutrient digestibility in food waste ingredients for Pekin and Muscovy ducks. AB - Food wastes are valuable resources to be recycled into new added-value products through animal production. The determination of energy and digestibility values of these wastes is essential for feed formulation. Corn, soybean meal (SBM), and a total of nine industrial food waste ingredients were tested in a comparative metabolic study in Pekin and Muscovy ducklings at two different ages during growth. The "precision-feeding" technique was employed to establish DM, fat, and fiber digestibility as well as retention of N and energy (AME, AMEn in Pekins; and AME, AMEn, TME, TMEn in Muscovies) for the 11 ingredients. For Pekin at 3 wk of age, the AMEn of peanuts, tofu, pogo, granola, waste diet, bread, corn, SBM, okara, and brewers grains were 5,141, 4,019, 3,971, 3,908, 3,141, 2,279, 1,572, and 1,442 kcal/kg, respectively. For Pekin at 6 wk of age, the AMEn of peanuts, pogo, tofu, granola, waste diet, bread, corn, SBM, and okara were 5,340, 4,327, 4,254, 4,079, 3,567, 3,302, 3,201, 2,416, and 1,562 kcal/kg, respectively. For Muscovy at 7 wk of age, the TMEn of peanuts, pogo, granola, waste diet, corn, tofu, bread, SBM, okara, and peanut skin were 5,207, 4,321, 4,057, 3,733, 3,233, 3,180, 3,084, 2,236, 1,575, and 904 kcal/kg, respectively. For Muscovy at 11 wk of age, the TMEn of peanuts, pogo, granola, tofu, waste diet, corn, bread, SBM, okara, and brewers grains were 5,077, 4,137, 4,025, 3,921, 3,586, 3,254, 3,123, 2,245, 2,007, and 1,392 kcal/kg, respectively. Nitrogen retention was significantly (P < 0.05) higher for SBM, tofu, okara, pogo, peanuts, and the food waste diet and lower for bread, corn, granola, brewers grains, and peanut skin. Dry matter digestibility was high for granola, pogo, corn, bread, and the food waste diet. Fat digestibility was generally the same for all the ingredients and was consistently over 97%. Bread neutral detergent fiber (NDF) was significantly (P < 0.05) the most digestible (88.92% NDF digestibility), as it consisted of 96.29% hemicellulose, whereas okara NDF was significantly (P < 0.05) the least digestible (26.94% NDF digestibility) and contained only 14.38% hemicellulose. Peanut skins and SBM with 30% hemicellulose showed only slightly higher digestibilities of NDF. The results of this study establish reliable data for formulation of duck diets using the tested industrial food waste ingredients as well as corn and SBM in both Pekin and Muscovy ducklings at two different ages during growth to market weight. PMID- 9733126 TI - Utilization of spent hen meal in diets for broiler chickens. AB - Studies were conducted to evaluate spent hen meal (SHM) produced in commercial rendering plants as a nutrient source in diets for broiler chickens. Utilizing previously determined nutrient composition values, including digestible amino acid and TMEn content, diets were formulated to include 0, 5, 10, and 15% of SHM from three different locations. In the first experiment, conducted in battery pens from 1 to 21 d posthatch, diets were formulated with digestible amino acid requirements set at 90, 95, or 100% of NRC (1994) total amino acid requirements. In the second experiment, conducted in floor pens from 1 to 49 d posthatch, diets were formulated with digestible amino acid requirements set at 95% of NRC (1994) total amino acid requirements. Samples of birds from the second experiment were processed to determine the possible influence of SHM inclusion on carcass yield. Results of the present studies indicate that SHM from commercial rendering facilities can be utilized in diets for growing broiler chickens provided adjustments are made in nutrient content and digestibility. When formulated on the basis of digestible amino acid content, levels of SHM up to 10% appear acceptable based upon body weight, feed conversion, bone ash, and carcass yield. Higher inclusion rates generally reduced performance. It is apparent that differences in nutritional quality may exist among products produced by different rendering facilities; however, evaluation of products to assess nutrient quality may be difficult under commercial conditions. As more information is generated regarding typical amino acid content and digestibility of rendered SHM, the product may be used with greater confidence in commercial diets. PMID- 9733127 TI - Bioavailability of iron in cottonseed meal, ferric sulfate, and two ferrous sulfate by-products of the galvanizing industry. AB - Iron depletion-repletion assays were carried out with young chicks to establish Fe bioavailability values for Fe2(SO4)3.7H2O (22.7% Fe), Fe-ZnSO4.H2O (20.2% Fe, 13.0% Zn), Zn-FeSO4.H2O (20.2% Zn, 14.2% Fe), and cottonseed meal (200 mg Fe/kg). Standard hemoglobin response curves were established using feed-grade FeSO4.H2O (28.8% Fe) or reagent-grade FeSO4.7H2O (20.1% Fe) as standards such that relative bioavailability (RBV) could be assessed for the experimental sources of Fe. Weight gain, hemoglobin, and hematocrit responded linearly (P < 0.05) to Fe supplementation in all assays. Using hemoglobin as the response criterion, slope ratio calculations established Fe RBV values of 126% for Fe-ZnSO4.H2O and 93% for Zn-FeSO4.H2O. The 126% value for Fe-ZnSO4.H2O was greater (P < 0.05) than the FeSO4.H2O standard (100%), but the 93% value for Zn-FeSO4.H2O was not different (P > 0.10) from the standard. However, evaluation of all criteria of response (hemoglobin, hematocrit, weight gain) suggested that neither Fe-ZnSO4.H2O nor Zn FeSO4.H2O had different Fe RBV values than FeSO4.H2O. Standard-curve calculations were used for assessment of Fe RBV in Fe2(SO4)3.7H2O and cottonseed meal, as only a single level of Fe addition was studied for each of these products. Iron RBV in Fe2(SO4)3.7H2O was estimated to be 37%, whereas Fe RBV in cottonseed meal was found to be 56%. Both of these values were lower (P < 0.05) than the FeSO4 standard. The data suggest that the two new products, representing combinations of FeSO4.H2O and ZnSO4.H2O by-products of the galvanizing industry, are excellent sources of bioavailable Fe, whereas ferric sulfate and cottonseed meal are relatively poor sources of usable Fe. PMID- 9733128 TI - Effect of acute heat stress on amino acid digestibility in laying hens. AB - An experiment was conducted to determine the effect of acute heat stress exposure on amino acid digestibility in laying hens. A total of 30 commercial laying hens were singly housed in an environmentally controlled facility, fed a standard laying ration, and exposed to a constant thermoneutral temperature (21 C) for 12 d. The hens were then randomly fed one of three diets (10 hens per diet) and exposed to three consecutive temperature periods (8 d each), which consisted of: 1) a constant 21 C temperature, 2) a cycling temperature of 35 C for 12 h and 29 C for 12 h, and 3) a constant 21 C temperature. The three isonitrogenous (18% CP) diets fed were: 1) a corn-soybean meal diet, 2) a corn-soybean meal diet containing 15% meat and bone meal, and 3) a corn-soybean meal diet containing 5% alfalfa meal and 20% wheat bran. Excreta were collected from all hens during the last 4 d of each temperature period and apparent amino acid digestibility was determined. There was a significant diet effect (P < 0.05) on amino acid digestibility. Digestibility of amino acids in Diet 2 (corn-soybean meal/meat and bone meal) was higher (P < 0.05) than in the other two diets. In addition, digestibility of amino acids in Diet 3 (corn-soybean meal/alfalfa meal/wheat bran) was significantly lower (P < 0.05) than in Diets 1 or 2. Heat stress generally had no significant effect on amino acid digestibility except for His and Lys digestibility. Histidine digestibility was higher during the heat stress period than during the initial and recovery thermoneutral periods, whereas Lys digestibility was higher during the heat stress period than during the initial thermoneutral period. These results indicated that acute heat stress (8 d) had no adverse effects on dietary amino acid digestibility in laying hens. PMID- 9733129 TI - D-allothreonine has no growth promoting efficacy for chicks. AB - One hundred and sixteen crossbred male chicks were used in two battery trials to establish the biological efficacy and toxicity of D-allothreonine (D-allo-Thr) relative to L-Thr. In the efficacy trial, graded doses of D-allo-Thr or L-Thr were added to a Thr-deficient (0.24% L-Thr) chemically defined diet and fed to chicks during the period 10 to 21 d posthatching. Addition of 0, 0.09, and 0.18% L-Thr produced marked linear (P < 0.01) growth and feed efficiency responses, but addition of 0.18 or 0.36% D-allo-Thr did not elicit a response in either weight gain or feed efficiency. In the toxicity trial, 2% D-allo-Thr or 2% L-Thr were added to a conventional 23% CP corn-soybean meal starter diet. During an 11-d feeding period, neither weight gain nor voluntary feed intake were affected (P > 0.10) by 2% additions of either compound. This experiment demonstrates that chicks cannot metabolize D-allo-Thr to L-Thr and that neither L-Thr nor D-allo Thr are growth depressing when provided in a large surfeit. PMID- 9733130 TI - Examination of antisense RNA and oligodeoxynucleotides as potential inhibitors of avian leukosis virus replication in RP30 cells. AB - Avian leukosis virus (ALV) is an economically important pathogen of chickens. Both antisense RNA and antisense oligodeoxynucleotides (ODN) have been used to diminish the replication and spread of other retroviruses. The use of antisense RNA and ODN to inhibit ALV replication has been examined in cultured RP30 cells. Using an expression system that constitutively transcribes antisense ALV RNA, one transfected cell clone showed a significant reduction in virus growth. However, this effect was not reproducibly observed in other transfected cell lines or in cells in which the antisense transcript was expressed from a regulatable promoter, even though a substantial amount of antisense transcript was generated. Antisense ODN complementary to several different target sites near the 5' end of the ALV genome were also tested for antiviral activity, by comparison of antisense ODN effects to those of randomized sequence controls. An antisense ODN complementary to the ALV primer binding site demonstrated a reproducible reduction in viral replication. However, when the corresponding region was specifically employed as a target for intracellular antisense RNA expression, there again was no significant inhibition of ALV. These results suggest that in vivo expression of antisense RNA is unlikely to be an effective way to generate transgenic poultry that are resistant to field strains of ALV. PMID- 9733131 TI - Isolation and characterization of a calpain activator in chicken skeletal muscle. AB - The calpains (E.C. 3.4.22.17) and calpastatin constitute an ubiquitous, intracellular, Ca2+-dependent protease/inhibitor system. This system has been implicated as a principal regulator of myofibrillar protein degradation in both ante-mortem and postmortem muscle. Although proteolytic activity of the calpains is primarily controlled through interaction of calpain and calpastatin, evidence for an activator(s) has been limited and the reported characteristics varied. The function of the activator has not been elucidated. A putative calpain activator has been isolated from the Pectoralis muscle of broiler breeders (Cobb x Cobb). The activator elutes from an ion-exchange column at approximately 200 mM NaCl. Addition of activator increased apparent m-calpain activity to a level demonstrating a fourfold increase in proteolysis. The activator/calpain complex maintains a requirement for Ca2+ for proteolytic activity. Under physiological conditions, presence of the activator negates the ability of calpastatin to inhibit m-calpain. Additionally, the activator alone does not demonstrate proteolytic activity. Effect of the activator is pH-dependent; in a physiological pH range, the activator enhances m-calpain proteolytic activity but at pH less than 6.75 the effect is to inhibit m-calpain. The activator's ability to modulate m-calpain activity and eliminate calpastatin's effect provides a further means of regulating this important enzyme system. PMID- 9733132 TI - Localization of macrophages in the ovarian follicles during the follicular growth and postovulatory regression in chicken, Gallus domesticus. AB - Macrophages may be a potential regulator of ovarian functions. The goal of this study was to determine the changes in the macrophage frequency (number of cells per unit square of tissue) during follicular growth, ovulation, and postovulatory follicular regression in chickens. Cryostat sections of ovarian stroma containing primary follicles, small white follicles, and preovulatory and postovulatory follicles of laying hens were immunostained for macrophage using mouse anti chicken macrophage monoclonal antibody. Macrophages were observed under a light microscope and counted by a computer assisted image analyzer. The frequency of macrophages in the theca layer was significantly greater in the small white follicles than in the primary follicles (P < 0.01) and also greater in the preovulatory follicles than in the small white follicles (P < 0.01). No significant differences were observed in the macrophage frequency between the third largest and largest preovulatory follicles. In the theca layer of postovulatory follicles, macrophage frequency was significantly greater than in that layer in the preovulatory follicles (P < 0.01); however, the frequency of macrophages decreased significantly in the Day 3 postovulatory follicles as compared with Day 1 postovulatory follicle (P < 0.05). These results suggest that macrophages may play an important role in the follicular development and regression of postovulatory follicles. PMID- 9733133 TI - Impedance recordings to determine change in extracellular volume in the brain following cardiac arrest in broiler chickens. AB - The present study describes a method to determine the onset and development of brain damage in broiler chickens. Exsanguination disrupts the brain metabolism and causes the brain to become ischemic. Energy-requiring systems in the cell membrane fail, which results in an ionic shift over the membrane, accompanied by a water influx into the cell. This cellular edema decreases the extracellular volume of brain tissue. In mammals, this brain damage has been measured by recording brain impedance. We adapted this approach for use with poultry. Five to six-week-old commercial broilers were equipped with impedance recording electrodes in the striatum area of the brain. Cardiac arrest was induced by means of an intravenous injection of MgCl2 and brain impedance was recorded for 30 min. The resulting curves showed a high similarity to those obtained in rats. No effects of 12 h antemortem feed deprivation on the size and rate of change in brain impedance could be found. Both in anesthetized and conscious birds, a change in brain impedance was found. We conclude that brain impedance can be used to determine the development of ischemic brain damage in broiler chickens. PMID- 9733134 TI - Effect of superoxide and superoxide-generating systems on the prooxidant effect of iron in oil emulsion and raw turkey homogenates. AB - Mechanisms of superoxide.O2--generating systems on the pro-oxidant effect of iron from various sources were studied. Reaction mixtures were prepared with distilled water, oil emulsion, or meat homogenates. Free ionic iron (ferrous and ferric), ferritin and hemoglobin (Hb) were used as iron sources, and KO2 and xanthine oxidase (XOD) systems were used to produce .O2-. Thiobarbituric acid reactive substances (TBARS) values and iron contents of the reaction mixtures were determined. Ferric iron and ferritin, in the presence or absence of superoxide generating systems, had no catalytic effect on the oxidation of oil emulsion but became pro-oxidants when reducing agent (ascorbate) was present. Ferrous iron and Hb had strong catalytic effects on the oxidation of oil emulsion as shown by TBARS values. Superoxide and H2O2, generated from superoxide-generating systems, oxidized ferrous iron and ascorbate, and lowered the pro-oxidant effect of ferrous iron in oil emulsion. Addition of ferric or ferrous iron increased but Hb did not have any effect on the TBARS values of raw meat homogenates. The reaction mechanisms of superoxide and the superoxide-generating systems on the prooxidant effect of various iron sources indicated that .O2- was a strong oxidizer rather than a reducing agent, and the antioxidant effect of XOD system in oil was caused by the oxidation of ferrous iron to the ferric form by .O2- and/or H2O2. PMID- 9733135 TI - Organoleptic evaluation of eggs produced by laying hens fed diets containing graded levels of flaxseed and vitamin E. AB - Eighteen-week-old laying hens were fed diets containing 0, 10, or 20% ground flaxseed and 10 or 100 IU vitamin E/kg diet, in a factorial arrangement. When birds were 60 wk of age, eggs were collected from various treatments and used in taste panel studies. All studies involved freshly boiled eggs. In Experiment 1, four separate panels were conducted to obtain information on egg aroma, egg flavor, presence of any off-flavor, and overall acceptability of the egg. Pooled data showed a very significant difference among panelists for all attributes tested (P < 0.01) and off-flavors were detected in eggs from hens fed 10 and 20% flaxseed with 10 mg vitamin E/kg. In Experiment 2 panelists were asked to evaluate egg aroma, yolk flavor, and overall acceptability in eggs from birds fed the lowest level of vitamin E with 0 or 10% flaxseed, and birds fed 20% flaxseed and 10 or 100 mg vitamin E/kg. The highest ratings for egg aroma, yolk flavor, and overall acceptability were for the control eggs. There was a significant reduction in overall acceptability as flaxseed concentration increased (P < 0.05) and an interesting and significant (P < 0.05) decline in overall acceptability for birds fed 100 vs 10 IU vitamin E/kg diet in eggs for birds fed 20% flaxseed. In a third study, there was an indication of preference for eggs from birds not fed flaxseed, when the diet contained 10, rather than 100 IU vitamin E/kg diet. These data suggest that high (> 10%) levels of flaxseed used in the bird's diet will result in some decrease in overall egg acceptability as assessed by aroma and flavor. These effects seem to be accentuated by using high levels of vitamin E in the bird's diet. PMID- 9733136 TI - Sensory and objective characteristics of broiler meat from commercial broilers fed rendered spent hen meal. AB - The objective of the study was to determine sensory and objective characteristics of broiler breast and thigh meat from commercial broilers fed rendered spent hen meal (RSHM) from hatch to 42 d of age. Breast and thigh muscles from 90, 6-wk-old straight-run broilers (i.e., mixed male and female broilers) fed starter and grower diets consisting of either 0, 8, or 12% RSHM were evaluated for sensory characteristics, instrumental texture, and compositional profiles. The RSHM treatments had no adverse effects (P > 0.05) on juiciness, chicken flavor intensity, tenderness, or compositional profiles for the breast or thigh meats. Off-flavor scores for all treatments were above the threshold value, indicating that the RSHM imparted no off-flavors to the breast and thigh meats. Warner Bratzler shear measurements were similar (P > 0.05) for breast meat from broilers in all treatments. No shear measurements were conducted for the thigh meat. It was concluded that RSHM can be incorporated into the diets of broilers at levels of up to 12% without causing objectionable sensory characteristics in the cooked broiler meat. PMID- 9733137 TI - Eggshell characteristics and penetration by Salmonella through the productive life of a broiler breeder flock. AB - Egg weight, specific gravity, conductance, and ability of Salmonella to penetrate the shell and membranes were determined for hatching eggs from a commercial broiler breeder flock. Thirty unsanitized eggs were sampled on Weeks 29, 34, 39, 42, 48, 52, and 56 of flock age for specific gravity and conductance. An additional 10 intact eggs were inoculated with Salmonella by a temperature differential immersion method for 1 min. Eggs were then emptied of contents and filled with a selective medium that allowed visualization of Salmonella growth on the inside of the shell and membrane complex. Over the 27-wk sampling period, egg weight increased from 56 to 66 g and was positively correlated with hen age (r = 0.96, P < 0.05). However, neither specific gravity (ranging from 1.077 to 1.082) nor eggshell conductance (ranging from 14.7 to 17.9 mg weight loss/d per torr) showed any clear trend throughout the life of the flock despite the increase in egg weight. Conductance values were not correlated with specific gravity. The number of eggs positive for Salmonella penetration after 24 h incubation showed a general upward trend with flock age; however, penetration frequency and hen age were not found to be significantly correlated (P > 0.05). No relationship was found between egg specific gravity, conductance, or egg weight and the likelihood of Salmonella to penetrate the eggshell. Because shell characteristics did not change over time and the penetration patterns did vary, it is likely that factors other than specific gravity and conductance were involved in the penetration of eggshells by Salmonella. PMID- 9733138 TI - Meat quality of broiler breast meat following post-mortem electrical stimulation at the neck. AB - This experiment was conducted to evaluate the effects of electrical stimulation (ES) on breast fillets harvested at 1 h post-mortem and individually quick frozen (IQF) or aged on ice (ICE). Twelve birds were electrically stimulated (450 V, 750 mA, 2 s on/1 s off for 15 s) at the neck in a saline bath. Twenty-four unstimulated birds were used as controls. Breast fillets were harvested at 1 h post-mortem from ES and control carcasses or at 4 h post-mortem from control carcasses and were either IQF or ICE until 24 h post-mortem. Fillets were then analyzed for shear value, pH, R value, and color. Electrical stimulation significantly reduced shear values compared to the 1 h controls for both IQF and ICE treatments. The ICE fillets deboned at 1 h from ES-treated carcasses had shear values similar to those of ICE fillets deboned from the 4 h controls. Electrical stimulation increased the percentage of shear values at or below 8 kg/g for the fillets from ES-treated carcasses compared to the 1 h controls. Electrical stimulation accelerated the normal post-mortem decline in pH and increase in R value. There was no significant difference in L* or a* values between the fillets held for 1 or 24 h. The results suggest that by electrically stimulating carcasses at the neck using a saline bath, the aging period could be eliminated by removing fillets immediately after chilling at 1 h, decreasing the costs associated with aging whole carcasses or front halves. PMID- 9733139 TI - SP22: a novel fertility protein from a highly conserved gene family. AB - Three nucleotide sequences encoding SP22, a protein originally identified in detergent extracts of cauda epididymal sperm, were isolated from a rat testis cDNA library. While two of these cDNA sequences differed only in their 5' untranslated regions, a third cDNA was predicted to contain an additional 13 amino acids of coding sequence. Amino acid sequences obtained following Edman degradation of purified SP22 protein and cDNA sequence data both indicated that SP22 was a member of a highly conserved and widely expressed gene family found in organisms as diverse as human and Escherichia coli. Interestingly, while a 1-kb mRNA transcript was widely expressed in somatic tissues, a unique pattern of testicular expression was observed, including the appearance of a novel 1.5-kb transcript and an increase in the abundance of the 1-kb transcript during spermatogenic cell development. Anti-SP22 peptide antiserum was shown to recognize a family of 22-kDa proteins on western blots of detergent-extracted cauda epididymal sperm protein, suggesting that multiple charge variants of SP22 coexist. Moreover, affinity-purified anti-SP22 peptide immunoglobulin localized in a highly specific manner to the anterior-ventral surface of the equatorial segment of the sperm head. This is an extremely intriguing finding as SP22 was originally shown to be highly correlated with, and predictive of, the fertilizing ability of cauda epididymal sperm. Although no conclusive function has been attributed to any members of the SP22 gene family, the localization of SP22 over a discrete region of the sperm head suggests a pivotal role in sperm-egg interactions. PMID- 9733140 TI - Interactions between testicular macrophages and Leydig cells. PMID- 9733141 TI - New classification system for erectile dysfunction therapies. PMID- 9733142 TI - Detection of human glandular kallikrein, hK2, as its precursor form and in complex with protease inhibitors in prostate carcinoma serum. AB - Forms of human glandular kallikrein (kK2) in prostate carcinoma serum were identified using monoclonal antibodies specific for hK2 and prohK2. Recombinant mammalian hK2, prohK2, and prostate = specific antigen (PSA) were utilized to confirm the specificity of monoclonal antibodies for hK2 and the lack of reactivity with PSA. In prostate cancer patient sera containing high levels of hK2 (>100 ng/ml), hK2 exists as a complex with alpha1-antichymotrypsin with a molecular weight of 90 kDa. The kallikrein also exists as a 32-kDa free form, which includes the precursor pro form of hK2. The relative amount of complex and free hK2 varied, but in most sera examined the 32-kDa form predominated. Recombinant hK2 readily formed complexes with alpha2-macroglobulin when the two proteins were incubated together as well as when hK2 was spiked into female serum. PMID- 9733143 TI - Antioxidant potential of human serum albumin: role in the recovery of high quality human spermatozoa for assisted reproductive technology. AB - Human serum albumin (HSA) is being considered as an alternate media for sperm enrichment in assisted reproductive technology (ART) because of recent concern with the use of Percoll. In this study, we compared HSA and Percoll for 1) sperm recovery, 2) reactive oxygen species scavenging potential, and 3) effects on total oxidative stress to spermatozoa. The spermatozoa-enriched fractions obtained from Percoll (80%:40%) and HSA (12%) were monitored for sperm motility, viability, hypoosmotic swelling test (HOST), and adenosine triphosphate (ATP) levels. The effect of superoxide anions (O2.-) on donor human spermatozoa was observed in the presence of either HSA or Percoll media. A combination of luminol and the Cypridina luciferin analog 2-methyl-6-(4-methoxyphenyl)-3,7 dihydroimidazo(1,2-alpha)pyraz in-3-one hydrochloride was used as a highly sensitive chemiluminescence probe in our hypoxanthine and xanthine oxidase-based assay for O2.-. Sperm membrane total oxidative stress was determined by measuring levels of the prostanoid 8-iso-Prostaglandin F2alpha (8-iso-PGF2alpha). Significant differences in sperm parameters between the Percoll-enriched spermatozoa (motility 60%+/-4%, viability 56%+/-6%, and HOST 73%+/-7%) and those enriched with HSA (motility 84%+/-5%, viability 85%+/-4%, and HOST 84%+/-3%; P < 0.01) were observed. Adenosine triphosphate levels were significantly higher, by almost 50%, in samples processed with HSA than with Percoll (P=0.03). The dismutation rate of O2.- in HSA (slope -6.8) was significantly lower than in Percoll (slope -87.0; P < 0.01). Sperm motility and ATP levels decreased at a slower rate after treatment with O2.- in the presence of HSA when compared to Percoll; moreover, spermatozoa in HSA regained partial motility after 2 hours, whereas spermatozoa in Percoll were immobilized. No significant differences in 8 iso-PGF2alpha levels in spermatozoa enriched by either HSA or Percoll were observed. We conclude that the HSA sperm enrichment procedure improves the recovery of higher quality spermatozoa compared to Percoll and, because of its antioxidant properties, may be useful in processing high leukospermia semen samples for ART purposes. PMID- 9733144 TI - FSH does not directly influence testicular macrophages. AB - We have previously demonstrated that conditioned medium from testicular macrophages stimulates testosterone production by Leydig cells. It was also reported that conditioned medium from macrophages treated with follicle stimulating hormone (FSH) had an even greater amount of Leydig cell-stimulating activity than medium from untreated macrophages, indicating that this factor is under the regulation of FSH. However, most other laboratories have been unable to reproduce this effect of FSH. We have recently purified and partially characterized the stimulatory factor from macrophage-conditioned medium that stimulates Leydig cells. The purpose of the present investigation was to reinvestigate the effect of FSH by determining whether it regulates the production of this purified factor and by determining whether macrophages have mRNA for the FSH receptor. Testicular macrophages were isolated from adult rats and incubated 24 hours with human recombinant FSH (20 units/ml), ovine FSH (200 ng/ml), fetal bovine serum (2%), or dibutyryl cyclic adenosine monophosphate (1 mM). The macrophage-derived factor (MDF) was then purified from conditioned medium of the various treatment groups and added to Leydig cells. The concentration of testosterone in the Leydig cell medium was then measured after 16 hours. It was found that serum significantly stimulated production of the MDF. However, FSH had no effect on production of the MDF in the presence or absence of serum. Dibutyryl cyclic adenosine monophosphate exerted a slight inhibitory effect on production of the macrophage-derived factor. Most importantly, testicular macrophages did not express detectable levels of FSH receptor mRNA, either in vivo or in vitro, when evaluated using either in situ hybridization or northern analysis, under identical conditions that clearly demonstrated FSH receptor mRNA in Sertoli cells. We conclude that testicular macrophages are not a direct target for FSH. PMID- 9733145 TI - Androgen concentrations and their receptors in the periurethral region are higher than those of the subcapsular zone in benign prostatic hyperplasia (BPH). AB - Benign prostatic hyperplasia (BPH) is an androgen-dependent disease that initially develops in the inner prostate, where the highest concentrations of testosterone (T) and dihydrotestosterone (DHT) are found. In this study, we have evaluated the cytosolic androgen receptors (ARc), the nuclear androgen receptors (ARn), and the concentrations of T, DHT, and 3alpha-androstanediol (3alphaDiol) in BPH tissue to verify the existence of a possible correlation between androgens and their receptor concentrations. Prostatic samples, removed by suprapubic prostatectomy in 15 untreated patients, were sectioned in periurethral, intermediate, and subcapsular zones. Testosterone, DHT, and 3alphaDiol were evaluated by radioimmunoassay after extraction and purification on celite microcolumns, and ARc and ARn were evaluated by means of dextran-coated charcoal method. In total tissue, mean levels of DHT, T, and 3alphaDiol were 2,531+/-308, 260+/-36, and 403+/-35 pg/mg of DNA (mean+/-SE), respectively. Cytosolic androgen receptors, detectable in all cases, were 16+/-2.8 fmol/mg of protein (mean+/-SE), and ARn, detectable in 12 cases, were 108+/-15 fmol/mg of DNA (mean+/-SE). A linear correlation between DHT and 3alphaDiol, T and DHT, and 3alphaDiol and ARn was found. If the different regions are considered, the periurethral zone, site of the primitive BPH nodule, presents the highest levels of androgens and ARn with respect to the other regions. This relative hyperandrogenism may be responsible for the growth-promoting processes of this area, leading to urinary obstruction. PMID- 9733146 TI - Interaction between Ca2+, cyclic 3',5' adenosine monophosphate, the superoxide anion, and tyrosine phosphorylation pathways in the regulation of human sperm capacitation. AB - In order to fertilize the egg, spermatozoa must go through the capacitation process where they experience Ca2+ uptake, increases in cyclic 3',5' adenosine monophosphate (cAMP) concentrations, superoxide anion production, and protein tyrosine phosphorylation. Although the importance of these processes has been described, the interactions between them, as well as the temporal sequence of these events, remain to be demonstrated. Previous studies from our laboratory have demonstrated that tyrosine phosphorylation of p105 and p81 (p105/81), the two major human sperm phosphotyrosine-containing proteins, was under cAMP and oxygen derivatives regulation. In the present study, we investigated the importance of intra- and extracellular Ca2+, as well as the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine and the phosphatase inhibitors calyculin A and okadaic acid, in the production of superoxide anion and p105/81 tyrosine phosphorylation. An increase in p105/81 phosphotyrosine content was observed when spermatozoa were incubated in the absence of extracellular Ca2+ or with the calmodulin antagonist N-(6-aminohexyl)-1-naphthalenesulfonamide. However, the human sperm capacitation inducer FCSu (ultrafiltrate of fetal cord serum) requires the presence of the extracellular Ca2+ to induce capacitation, superoxide anion production, and tyrosine phosphorylation of p105/ 81, whereas free intracellular Ca2+ had no effect on these last two processes. The production of superoxide anion by spermatozoa was stimulated by inhibitors of phosphodiesterases and serine/threonine phosphoprotein phosphatases. The tyrosine phosphatase inhibitor vanadate decreased by 40% the FCSu-stimulated superoxide anion production, although it had no effect when used alone. These results suggest that, during sperm capacitation, Ca2+ induces an elevation in cAMP levels; this cAMP, through undefined serine/threonine protein phosphorylation, stimulates the generation of superoxide anion, which, in turn, causes the increase in p105/81 phosphotyrosine contents. PMID- 9733147 TI - Deficiency of fucosidase results in acrosomal dysgenesis and impaired sperm maturation. AB - Although a variety of glycosyltransferases and glycosidases have been implicated in spermatogenesis and posttesticular sperm maturation, the biological role of these enzymes in these processes is largely unknown. We describe reproductive sequelae in a cohort of male dogs suffering from fucosidosis, a heritable lysosomal storage disorder caused by a severe deficiency of alpha-L-fucosidase. There was a reduction in the total number of sperm in the ejaculate. Only 3-5% of sperm were motile. None of the sperm were found to be morphologically normal. The predominant morphological defects observed were malformed acrosomes (56%) and retained proximal cytoplasmic droplets (92%), indicating that spermiogenesis and sperm maturation were impaired. The cytoplasm of all cellular components of the testis and excurrent ducts were vacuolated. The vacuolation resulted from enlargement of lysosomes caused by accumulation of compounds that are otherwise cleaved/degraded when lysosomal hydrolases are present normally. It is possible that impairment in spermatogenesis, particularly morphogenesis of the acrosome, is due to physical damage caused by anomalous enlargement of lysosomes. Although an unambiguous causal relationship could not be established, it is evident from the available information that the derangement in events associated with epididymal sperm maturation, namely acquisition of motility and shedding of the cytoplasmic droplet, is likely due to lack of fucosidase leading to impaired sperm membrane modification. This heritable condition in dogs may serve as a spontaneously occurring knock-out model for further elucidating the role of alpha L-fucosidase in spermatogenesis and sperm maturation. PMID- 9733148 TI - The effects of aging on the expression of glutathione S-transferases in the testis and epididymis of the Brown Norway rat. AB - Glutathione S-transferases (GSTs), a family of isoenzymes, catalyze the conjugation of glutathione to a variety of electrophiles, and protect cellular constituents from electrophilic and oxidative attack. Aging is associated with an overall increase in oxidative stress and thus free radical production. The present study examines the immunocytochemical localization of Ya, Yc, Yb1, Yb2, Yo, and Yf GST subunits in the testis and epididymis of Brown Norway rats aged 3, 12, 18, and 24 months. In the testis, neither Sertoli nor germ cells showed changes in the GST staining pattern during aging. At 24 months, two types of Leydig cells were noted. Some (peritubular) formed a distinct band at the periphery of the tubule while others were seen in the interstitial space. The peritubular cells were identified as Leydig cells by specific staining for 3beta hydroxysteroid dehydrogenase (3beta-HSD), a Leydig cell-specific marker. Both types of Leydig cells were intensely reactive for all GST subunits at all ages. In the epididymis, principal cells of all epididymal regions, except the proximal cauda region, showed no changes in GST expression at all ages examined. At 24 months, some principal cells of this region became greatly enlarged and vacuolated. These cells were unreactive for Yo, Yb1, Yb2, and Yc, while adjacent normal-appearing principal cells maintained the same intensity of expression as seen in 3-month controls. In contrast, vacuolated principal cells were reactive for the Ya subunit, while adjacent normal principal cells were unreactive. These data indicate that selective changes occur in the expression of GSTs at 24 months in principal cells having both a normal and a vacuolated appearance. The underlying mechanism responsible for these changes with age is unresolved, but we speculate that they lose the ability to handle oxidative stress. Taken together, these data show that aging affects region-specific changes in GST expression in the epididymis and Leydig cell distribution in the testis. PMID- 9733149 TI - Patterns of efferent lymphatics of the mouse testis. AB - Previous studies in the rat demonstrated that lymph vessels arise from various sites in the testis and then anastomose to form the main lymph trunk toward the testicular artery. Although mice are as commonly used in the laboratory as the rat, precise knowledge concerning their testicular lymphatics has not been established. In the present study, the patterns of testicular lymphatic drainage in the mouse were examined using local injection of India ink into the testicular parenchyma. The results demonstrated that, besides the lymph trunk surrounding the testicular artery, a lymphatic drainage route running along the ductus deferens is also present. PMID- 9733150 TI - The loss of alpha-adrenergic effect during the erectile response in the long-term diabetic rat. AB - The present study was designed to investigate the effect of long-term, streptozotocin-induced diabetes on the erectile response in the laboratory rat. Mean arterial blood pressure (MAP) and intracavernosal blood pressure within the erectile tissue (CCP) were continuously monitored during erection elicited by stimulation of the autonomic innervation of the penis. MAP and CCP were also measured during administration of two drugs: nitroglycerin, a nitric oxide donor drug and phenylephrine, an alpha-adrenergic agonist. The results of these studies show that during graded electrical stimulation of the ganglion, the overall magnitude of the erectile response was greater in the diabetic rats than in untreated control animals. Neither diabetic nor control animals responded significantly to infusion of nitroglycerin. However, diabetic rats and control rats responded very differently to administration of phenylephrine; in the control rats, this alpha agonist caused a sharp decline in CCP as the cavernosal vessels constricted in response to the drug. The same dose of phenylephrine had no discernible effect on CCP in the diabetic animals. This loss of alpha responsiveness may be confined to the penile circulation because MAP was elevated to approximately the same extent in both groups. Taken together, these results show that long-term diabetes leads to a failure of alpha-adrenergic responsiveness in the cavernosal circulation. The greater erectile response to ganglionic stimulation in the diabetic animals is likely due to the loss of response to endogenous norepinephrine. PMID- 9733151 TI - Triptolide: a potential male contraceptive. AB - The antifertility effect of triptolide and other related compounds, isolated from Tripterygium wilfordii, has been demonstrated in male rats. The exact sites and mechanism of action of triptolide remain unknown. Our objectives were to determine whether triptolide at selected dose levels that induce infertility has any detrimental effects on the testes and to determine the sites and the possible mechanisms of its action. Groups of six adult male Sprague-Dawley rats were given oral administration of either vehicle (control group) or triptolide (50 or 100 microg/kg body weight) daily for 35 or 70 days. Body weight gain was normal in all treated groups. All six rats treated with a high dosage of triptolide were infertile during the second (63-70 days) mating trial. A lower dose (50 microg) of triptolide gave intermediate fertility values. Plasma levels of luteinizing hormone, follicle-stimulating hormone, testosterone, and intratesticular testosterone were not significantly different between control and triptolide treated groups. Cauda epididymal sperm content was decreased by 68% and the motility, which averaged 58.2% in the control rat, was reduced to almost zero. No effects of triptolide were observed on testis and accessory organs weight, volumes of tubular lumen and the total Leydig cells, tubule diameter, and the number of Sertoli cells, spermatogonia, preleptotene (PL), and pachytene (P) spermatocytes. There were, however, modest but significant decreases in tubule volume and the number of round spermatids at stages VII-VIII. No changes in the germ cell apoptotic index measured at stages VII-VIII and XIV-I were noted between controls and rats rendered infertile with a high dose of triptolide. Thus, triptolide, at a dose level that induces complete infertility in the adult rats, has minimal adverse effects on the testes and acts primarily on the epididymal sperm making triptolide an attractive lead as a post-testicular male contraceptive. PMID- 9733152 TI - Apoptosis pattern elicited by several apoptogenic agents on the seminiferous epithelium of the adult rat testis. AB - Spontaneous germ cell death during spermatogenesis is an important event, and the usefulness of the seminiferous epithelium as an in vivo model to study apoptosis has been evidenced. Nevertheless, the response of the testis to apoptogenic agents has not been analyzed. This study was designed to determine germ cell sensitivity to induction of apoptosis and to provide baseline data on the testis response to several apoptogenic agents. Induced apoptosis was assessed by in situ DNA 3'-end labeling and quantified at every stage of the spermatogenic cycle. The shortest response time for every agent was established based on morphological and quantitative criteria. Our results show significantly increased incidence of germ cell deaths after all treatments, mainly at stages I, XII, and XIV. These specific stages coincide with those at which the greatest numbers of spontaneous germ cell deaths occur in control animals. Moreover, the rapid and highly specific response of germ cells to all the apoptogenic agents used in the present study indicate that apoptosis must be tightly regulated at these stages of the seminiferous epithelium. As a consequence, we propose that the disruption of apoptosis control might be an important determinant for idiopathic male infertility. PMID- 9733153 TI - Effect of medium-term supplementation with a moderate dose of n-3 polyunsaturated fatty acids on blood pressure in mild hypertensive patients. AB - Several studies have shown that n-3 polyunsaturated fatty acids (n-3 PUFA) are able to lower blood pressure (BP) in humans, but large doses of fish oils have been often used. Moreover, most of the studies available in the literature were not able to evaluate the specific effects of n-3 PUFA because they employed fish oils which contain, together with n-3 PUFA, many other different components. The aim of this preliminary study was to evaluate if medium-term supplementation with a moderate dose of highly purified eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) ethyl esters is able to reduce BP in mild hypertensive patients. Sixteen mild essential hypertensive (diastolic BP: 95-104 mm Hg), non diabetic, normolipidemic male outpatients and 16 normotensive male controls were recruited to participate in the study. Both hypertensive and control subjects were randomly assigned to receive either EPA and DHA ethyl esters (2.04 g EPA and 1.4 g DHA) as active treatment or olive oil (4 g/day) as a placebo for a period of 4 months. These subjects were followed up with 24-hour ambulatory BP monitoring and blood chemistry analyses at 2 and 4 months of treatment and 2 months after its discontinuation. The intake of n-3 PUFA was checked by red blood cell (RBC) phosphatidylcholine (PC) fatty acid composition. The effect of n-3 PUFA on BP in the active group was maximum after 2 months. Both systolic (-6 mm Hg, p<0.05) and diastolic (-5 mm Hg, p<0.05) BP significantly decreased during the n-3 PUFA ethyl ester supplementation. No further effect was observed at 4 months with a return to baseline values during the recovery period. These data indicate that 4 g/day of highly purified EPA + DHA ethyl esters are able to favorably affect BP in mild hypertensives. PMID- 9733154 TI - S-nitrosoglutathione/glutathione disulphide/Cu2+-dependent stimulation of L arginine transport in human platelets. AB - In this study, we have examined the effects of authentic nitric oxide (NO), NO+ (NOBF4), glutathione (GSH), glutathione disulphide (GSSG), and S nitrosoglutathione (GSNO) in the presence and absence of Cu2+, which thermally releases NO from S-nitrosothiols on the transport of L-arginine into the human platelet. The K(M,apparent) was unaffected by NO, NO+, GSH, and GSNO. However, Cu2+ lowered K(M,apparent) by approximately 2.85-fold. Cu2+-dependent lowering of K(M,apparent) was also observed, albeit to a smaller extent when this ion was mixed with GSH (approximately 1.9-fold lower) and GSNO (approximately 2.0-fold). GSSG also lowered K(M,apparent) by approximately 1.5-fold. The Vmax,apparent of L arginine uptake was unaffected by NO, NO+, GSH, and Cu2+. Vmax,apparent was stimulated by to the largest extent by GSNO (approximately 2.28-fold) and GSNO plus Cu2+ (approximately 2.7-fold). GSSG and GSH plus Cu2+ also increased Vmax,apparent by approximately 1.9-fold. When these parameters are expressed in terms of transport efficiency (Vmax/K(M)) the largest effect of nearly 4.7-fold (over controls) was obtained by a combination of GSNO plus Cu2+. These results suggest that platelet L-Arg transport is not affected either by NO or NO+ but by a thiol-disulphide exchange reactions on the platelet L-Arg transporter, brought about by GSNO and GSSG. Based on these results, a GSNO/GSSG/Cu2+ dependent regulatory mechanism for the uptake of L-arginine in human platelets has been proposed. PMID- 9733155 TI - Prothrombin fragment 1 + 2 measures treatment effect in patients with antiphospholipid syndrome. AB - Antiphospholipid syndrome (APS) is characterized by recurrent thrombosis. The anticoagulant management of APS thrombosis remains controversial. Few reports on markers of in vivo activation of coagulation have been reported. To determine whether plasma levels of prothrombin fragment 1 + 2 (F1 +2) correlate with thrombotic risk and treatment effect in patients with APS, plasma F1 + 2 levels were followed in 57 patients with this syndrome for more than 2 years. Clinical findings were also observed in these patients. Plasma levels of F1 + 2 in patients with APS were significantly higher when compared with control subjects (p<.05). These results suggest patients with APS are in a hypercoagulable state. Plasma levels of F1 + 2 significantly decreased following treatment with either aspirin, or aspirin plus warfarin (p<.05 and p<.01, respectively). Recurrent thromboses or spontaneous abortions occurred in all eight patients whose plasma levels of F1 + 2 remained higher than 1 nmol/l after treatment with either aspirin alone or no anticoagulants. These patients were subsequently treated with warfarin as well as aspirin, and plasma levels of F1 + 2 decreased to less than 1 nmol/l, with no additional thrombotic events over the remainder of the 2-year follow-up. No fatal bleeding was observed in treated patients. Our results suggest plasma levels of F1 + 2 are useful indicators of successful treatment. It is also suggested that warfarin plus mini-dose aspirin therapy is effective for patients with APS to protect from recurrent thromboses without harmful side effects. Further, prospective cohort studies are needed to substantiate these associations. PMID- 9733156 TI - Characterization of the binding of FK633 to the platelet fibrinogen receptor. AB - 125I-fibrinogen bound to ADP-activated fixed platelets in a saturable manner. The Scatchard plot was curvilinear but nonlinear model fitting of the data suggested that there was only one binding site with KA=4.19+/-1.3 x 10(6) M(-1) and a maximum number of binding sites of 3.9+/-1.1 x 10(4) molecules/platelet. The GPIIb/IIIa antagonists RGDS and FK633 both inhibited 125I-fibrinogen binding (50 microg/ml) in a dose-dependent manner. FK633 had a KB value of 2.5+/-0.48 x 10(7) M(-1) (Kb=39.9 nM) and an IC50, value of 64 nM, while RGDS had an KB of 2.55+/ 0.76 x 10(4) M(-1) (Kb=39.2 microM) with an IC50 value of 63 microM. At concentrations below its IC50 value FK633 was a competitive antagonist of fibrinogen binding. However, at concentrations above its IC50 value it was a noncompetitive antagonist. The IC50 values of FK633 remained constant over a wide range of fibrinogen concentrations while its KB value changed-increasing from 1.8+/-0.6 x 10(7) M(-1) at 10 microg/ml 125I-fibrinogen to 4.5+/-1.6 x 10(7) M( 1) at 300 microg/ml and decreasing to 0.3+/-0.2 x 10(7) M(-1) at 2.4 mg/ml. Thus, FK633 is a reversible, noncompetitive antagonist of fibrinogen binding to the platelet GPIIb/IIIa receptor. PMID- 9733157 TI - Body weight does not predict for anti-Xa levels after fixed dose prophylaxis with enoxaparin after orthopedic surgery. AB - Enoxaparin after joint arthroplasty is effective prophylaxis against venous thromboembolism. This is usually given as a fixed dose without monitoring of anti Xa levels. This study assesses the relationship between trough anti-Xa levels, body weight, and venous thromboembolism. Consenting patients at three institutions were treated with Enoxaparin 30 mg subcutaneously bis in die postoperatively until discharge. Chromogenic anti-Xa levels were measured on the fifth postoperative day by the method of Stachrome (Diagnostica Stago). All patients had bilateral compression doppler ultrasonography on day 10 or discharge and were followed for 12 weeks for evidence of venous thromboembolism. Eleven patients developed objectively confirmed venous thromboembolism during the study. In this study, there was poor correlation between weight and anti-Xa levels. In addition, body weight and anti-Xa levels of patients who developed venous thromboembolism were compared to those who did not and there were no significant differences between the two groups. In conclusion, this study shows that there is poor correlation of trough anti-Xa levels with body weight. Recognizing the low overall event rate this study does not support the need to monitor anti-Xa levels or adjusting the dose according to weight. PMID- 9733158 TI - Neutralase reverses the anti-coagulant but not the anti-thrombotic activity of heparin in a rabbit model of venous thrombosis. AB - Neutralase (heparinase I; E.C. 4.2.2.7) is a heparin-degrading enzyme undergoing clinical evaluation as an alternative to protamine for reversing the anticoagulant effects of heparin in coronary bypass surgery. The objective of this study was to assess the relative effects of Neutralase and protamine on reversal of heparin-dependent elevations in coagulation parameters and inhibition of clot formation in a rabbit vena caval stasis model. Rabbits were treated with saline or heparin (300 U/kg) for 10 minutes, followed by saline, protamine (2.6 mg/kg), or Neutralase (10 or 30 microg/kg, representing 1.23 IU/kg and 3.69 IU/kg, respectively). Twenty minutes later, venous stasis was induced, and vena caval clots were excised, weighed, and characterized. Coagulation parameters [activated partial thromboplastin time (aPTT) and thrombin clotting time (TCT)] and antiFactor IIa and Xa levels were measured throughout the protocol. Both protamine and Neutralase reversed heparin-mediated increases in aPTT (>300 seconds to 26-35 seconds) and TCT (>300 seconds to 29-56 seconds) to values that were not different from saline-treated, nonheparinized animals. Thrombus weight in the nonheparinized saline group was 62+/-7 mg; heparin-treated animals had no detectable clots. Protamine reversal of heparin was associated with clot formation (89+/-20 mg) while Neutralase reversal was not (no clots). Heparin induced increases in antiFactor IIa activity were reversed similarly by protamine and Neutralase (from 4.3-8.8 U/ml to 0.2-0.3 U/ml) while antiFactor Xa activity was differentially reversed (from 3.9-5.9 U/ml to 0.7-1.3 U/ml Neutralase; 5.5 U/ml to 0.02 U/ml protamine). These results are consistent with a hypothesis that Neutralase cleaves heparin into fragments, which are devoid of antiFactor IIa activity that retain modest antiFactor Xa activity, resulting in reversal of anticoagulant, but not antithrombotic, heparin activity. This property of Neutralase may be beneficial in reducing post-surgical thrombotic events after reversal of heparin. PMID- 9733159 TI - Low-dose heparin pretreatment is not able to prevent clotting activation during coronary angiography. PMID- 9733161 TI - Neurofibrillary degeneration and cell loss in the nucleus basalis in comparison to cortical Alzheimer pathology. AB - Neurofibrillary tangle staging was compared in the nucleus basalis and cerebral cortex of Alzheimer's disease patients with and without Lewy body disease. In pure Alzheimer's disease, cholinergic nucleus basalis cell number, as determined from counts in serial forebrain sections, was 22-60% of control mean, with the majority of residual cells containing tangles. A comparison between control cell number and the combined number of tangles plus tangle-free neurons in pure Alzheimer's disease suggests that the majority of nucleus basalis neurons were lost through neurofibrillary degeneration. The staging of neurofibrillary degeneration in the nucleus basalis was discordant with cortical changes as some controls had more extensive tangle formation in the nucleus basalis than in the cerebral cortex. Patients having both Alzheimer's disease and Lewy body pathology had few or no tangles in the nucleus basalis despite greater loss of neurons than purely demented patients. The presence of concomitant pathology had a greater effect on nucleus basalis tangle burden than did cortical disease stage, suggesting dichotomous disease processes in the cerebral cortex and forebrain. PMID- 9733160 TI - Mitotic phosphoepitopes precede paired helical filaments in Alzheimer's disease. AB - We have shown previously that the TG-3 and MPM-2 antibodies recognize phosphoepitopes common to mitosis and degenerating neurons of Alzheimer's disease(AD) brain. Here, we have evaluated their occurrence in human brain biopsy tissue, and confirm that they are absent in mature neurons of adult brain, but reappear during neurodegeneration in AD. The TG-3 epitope appears ahead of the MPM-2 epitope and is distributed throughout the neuronal soma. Tau is the major TG-3 antigen in AD brain. The initial localization of MPM-2 immunoreactivity in primary dendrites, it's robust occurrence in granulovacuolar bodies, and the increased immunoreactivity with 300-350-kDa proteins, suggest MAPI B as a candidate MPM-2 antigen in AD. Production of mitotic phosphepitopes in more than one type of human neurodegenerative lesion implicates mitotic kinases as common mediators of neuronal death. Because mitotic phosphoepitopes appear before paired helical filaments, it is suggested that mitotic kinase activation triggers neurofibrillary tangle formation. Future studies will need to focus on factors influencing mitotic kinase activity, a point with potential for early diagnosis and disease abrogation. PMID- 9733162 TI - Cerebrospinal fluid production is normal in Down syndrome. AB - The rate of production of cerebrospinal fluid (CSF) and the caudorostral gradients of total CSF protein were measured in seven subjects with Down syndrome (DS) and compared to age-matched healthy normal volunteers. The CSF production rate in DS subjects (0.35 +/- 0.02 mL/min.) did not differ significantly from normal subjects (0.37 +/- 0.09 mL/min.). In addition, the caudorostral gradient of total protein was similar in DS and normal subjects, with more caudal fractions of lumbar CSF having higher total protein levels than more rostral fractions. These data suggest that there is no gross disturbance in CSF dynamics in DS. PMID- 9733163 TI - Attenuation of age-dependent oxidative damage to DNA and protein in brainstem of Tg Cu/Zn SOD mice. AB - Age-dependent accumulation of oxidative DNA and protein damage in brainstem and striatum was assessed in normal and transgenic (tg) mice which overexpress human Cu/Zn superoxide dismutase (h-SOD1). A marker of oxidative DNA damage, 8-hydroxy 2'-deoxyguanosine (oxo8dG), was measured at 3, 12, and 18 months of age in control and tg mice. Cu/Zn SOD, but not MnSOD, activities in brainstems and striata from tg mice were increased compared to controls at all ages. At 18 months, oxo8dG levels were increased by 58% in brainstem and by 21% in striatum of control mice. In the tg mice, brainstem and striatal oxo8dG levels were increased to a lesser extent than in the corresponding controls. Protein oxidation (carbonyl content), was increased by 59% at 18 months in control brainstem, but not in striatum, and the increase was significantly attenuated in the tg mice. In summary, oxidative damage to DNA and protein increased with age in brainstem (and to a lesser extent in striatum), and augmented Cu/Zn SOD activity modified the extent of DNA and protein damage. PMID- 9733164 TI - Age-dependent local modulation of hippocampal-evoked responses to perforant path stimulation. AB - Local modulation of hippocampal-evoked responses to perforant path stimulation was studied by leaking drugs from the recording pipette placed in the dentate gyrus of anesthetized young (3 months old), aging (17 months old) and old (28 months old) rats. In old rats, the excitatory postsynaptic potential (EPSP) slope was much reduced compared to young and aging rats. The population spike (PS) size was similar in all age groups. Bicuculline caused a marked increase in PS size relative to population EPSP, and reversed the response to the second pulse in a paired-pulse paradigm from inhibition to facilitation. The effect of bicuculline was only slightly reduced in old rats. The 5-HT1a agonist 8-OH-DPAT potentiated PSs in the dentate gyrus, while not affecting paired-pulse inhibition. The effect of 8-OH-DPAT was slightly reduced in old rats. Carbachol, a cholinergic agonist, reversed paired-pulse inhibition into facilitation in the young brain, but not in aging and old rats. These results demonstrate that age affects differentially the action of biogenic amines on hippocampal reactivity to afferent stimulation. PMID- 9733165 TI - Age-related changes in the volume of somata and organelles of cerebellar granule cells. AB - Because cerebellar granule cells are fixed post-mitotic cells, it is expected that they undergo age-related changes like other neurons. To examine this possibility, a stereological study on granule cells of rat neocerebellar cortex was performed for an age spectrum of 2 to 24 months using eight different age groups. The nucleator method, together with point and intersection counting, was used to obtain primary data; arithmetical calculations determined the secondary data. In the soma, the absolute surface area did not change significantly; the volume did, however, exhibit a significant negative linear trend with age. Excluding dense bodies, the absolute volumes of the cytoplasmic components did not vary significantly. The absolute volume of dense bodies displayed a significant positive linear trend with age. Significant positive correlations were detected between the somatic volume and the absolute volume of either mitochondria or ground substance. It was concluded that granule cells showed a fair degree of morphological stability through 18 months. However, the observed changes warn that accompanying physiological alterations may occur, with putative effects on motor coordination. PMID- 9733166 TI - Uninjured aged sympathetic neurons sprout in response to exogenous NGF in vivo. AB - The extent to which the loss of plasticity by aged neurons is due to changes in the neuronal environment or to a loss of growth potential of the neurons has not been determined. In previous studies we observed that young adult cerebrovascular axons undergo a sprouting response following a 2-week intracerebroventricular infusion of nerve growth factor (15 microg; NGF). The present study used electron microscopy to examine the innervation of the intradural segment of the internal carotid artery of the aged rat and to determine whether aged sympathetic perivascular axons would respond to in vivo infusion of NGF. Young adult and aged Fischer 344 female rats received a 2-week intracranial infusion of NGF (15 microg) or vehicle (VEH) and were perfused for electron microscopy. Although there was no change in the total number of perivascular axons associated in aged VEH when compared with young adult VEH, a significant reduction was observed in aged VEH when total axons and sympathetic axons were expressed per microm2 vascular wall, reflecting an age-related increase in blood vessel size. Following NGF infusion, aged sympathetic axons were significantly increased by 192% compared with aged VEH cases. These results suggest that there is a proportional reduction in sympathetic cerebrovascular neurons with aging but that they exhibit robust sprouting in response to an exogenous neurotrophin. PMID- 9733167 TI - Multiple synaptic connections of a single neuron change differentially with age. AB - The efficacy of chemical synaptic connections of a single identified interneuron with different types of follower neurons was studied throughout the adult life of the pond snail Lymnaea stagnalis. Simultaneous intracellular recordings were made from the interneuron RPeD1 and its follower neurons in isolated CNS preparations from animals of different age groups (3-18 months of age). The presence of postsynaptic responses to RPeD1 action potentials was tested. With increasing age, the number of A-group neurons that was found with a response to evoked RPeD1 action potentials decreased, yet the number of HIJK-group neurons responding to RPeD1 input increased. The number of G-group neurons and the number of individual neurons VD2/3 and VD4 with RPeD1 input did not differ significantly between age groups. However, there was variability in the presence of responses in these individual neurons. Thus, synaptic connections of the single interneuron RPeD1 change differentially throughout the adult life of L. stagnalis. Within the A group we found indications that changes in RPeD1 input apply to the entire A group. In the A-group neurons changes in several electrical properties could not account for the observed age-related changes in the number of neurons responding to RPeD1 action potentials. PMID- 9733168 TI - Septo-hippocampal cholinergic and neurotrophin markers in age-induced cognitive decline. AB - Messenger RNA (mRNA) molecules encoding proteins related to the presynaptic cholinergic and neurotrophin systems were quantitated in the hippocampus and basal forebrain of Long-Evans rats with spatial learning ability assessed in the Morris water maze. The reverse transcriptase-polymerase chain reaction showed that the mRNAs for the low-affinity neurotrophin receptor (p75-NTR) and the growth-associated protein GAP-43 were decreased in level in the basal forebrain of aged-impaired rats. In the hippocampus of these aged-impaired rats, the mRNA for VGF, another neurotrophin-inducible gene, also was decreased. In situ hybridization histochemistry revealed that mRNAs for nerve growth factor (NGF) and brain-derived neurotrophic factor increased in level in the aged rat hippocampus; when age effects were removed, NGF mRNA level remained significantly correlated with maze performance. Enzyme-linked immunosorbent assay indicated that NGF protein was expressed at normal levels in the aged rat hippocampus. These mRNA and protein alterations may signify that a defect in neurotrophin signaling exists in the brains of aged Long-Evans rats, underlying reduced plasticity responses in the basal forebrain cholinergic system. PMID- 9733169 TI - Age-related levels of urinary free cortisol in the tree shrew. AB - There are still controversies concerning the effect of aging on the basal glucocorticoid concentration in mammals, including humans. Some studies reported an elevated glucocorticoid concentration in older subjects, while other reports showed no increases with age. These discrepancies may be caused by different experimental designs, gender differences, or varying sampling time points. The bulk of animal studies reporting increases of glucocorticoids with age were performed in rats. The present study was designed to investigate the impact of age on adrenocortical activity in a non-rodent mammalian species, tree shrews (Tupaia belangeri). We analyzed the basal urinary free cortisol concentration in the morning urine of male tree shrews in different age groups. Immediately after birth, a large variation in basal urinary free cortisol concentration (10-818 pg/micromol crea) has been observed. Between 21-40 day of age, the urinary cortisol concentration was low (32.7 +/- 5.6 pg/micromol crea) and increased steadily during puberty until adulthood (201-500 days; 161.8 +/- 15.1 pg/micromol crea). Thereafter, no further rise in basal urinary free cortisol concentration was found with increasing age and after reaching senescence (7-8 years). PMID- 9733170 TI - Effect of maternal hydration on mild fetal pyelectasis. AB - The aim of our study was to determine whether maternal hydration status prior to prenatal sonography affects fetal renal pelvic diameter. The renal pelvic diameters of fetuses from two different institutions were compared prospectively. At one institution 74 women were asked to drink 32 to 48 ounces of water prior to undergoing sonography (hydration group), whereas at the second institution, no specific hydration regimen was requested of 176 subjects. The inclusion criteria were as follows: greater than 15 weeks' gestation, otherwise normal obstetrical sonogram, normal amniotic fluid volume, and negative family history for renal disease. Renal pelvic diameter, degree of maternal bladder fullness, and gestational age were compared between the two groups using logistic regression analysis and log-linear analysis. A P value < 0.01 was considered significant. Bladder fullness in the two groups differed significantly (P < 0.001). Logistic regression analysis showed a very strong effect of maternal bladder fullness on fetal renal pelvic diameter (P < 0.001). The log-linear analysis model showed a highly significant association between maternal bladder fullness and fetal renal pelvic diameter (P < 0.001). We conclude that maternal hydration influences fetal renal pelvic diameter. The larger fetal renal diameters seen in the hydrated group support physiologic theories that the effects of maternal hydration on amniotic fluid volume are partially mediated via fetal urine production. PMID- 9733171 TI - Resolution of human parvovirus B19-induced nonimmune hydrops after intrauterine transfusion. AB - Our objective is to report our experience with cases of prolonged recovery from nonimmune hydrops secondary to human parvovirus B19 infection occurring after intrauterine transfusion. We reviewed cases referred to our unit over a 10 year period for exposure to parvovirus B19 infection. Those cases with serologic evidence of recent infection were identified. The cases requiring intrauterine transfusion were reviewed for demographic details, time of exposure, parvovirus B19 serology, gestational age at detection of nonimmune hydrops, number and results of fetal blood samples, duration from intrauterine transfusion to resolution of hydrops, and neonatal outcome. Of 38 cases identified through serologic evidence of recent parvovirus B19 infection, 35 (92.1%) did not develop hydrops, and these were followed by serial ultrasonography for 8 weeks from the time of exposure. Three cases (7.9%) developed hydrops and required intrauterine transfusion; in two the transfusion was intravascular and in one it was intraperitoneal. The mean duration from intrauterine transfusion to resolution of hydrops was 8 weeks 2 days. Pregnancy outcome in all cases was normal. In cases of nonimmune hydrops secondary to parvovirus B19 infection, resolution of the hydrops after intrauterine transfusion may take up to 12 weeks with a normal pregnancy outcome. PMID- 9733172 TI - Sonographic features of ovarian remnants. AB - Ovarian remnants occur after a portion of ovarian tissue is left behind unintentionally after oophorectomy. The ovarian remnant may be functional and cystic, producing pelvic pain and, in some patients, extrinsic compression of the distal ureter. Ovarian remnants frequently are associated with adhesions from previous pelvic surgery for endometriosis or pelvic inflammatory disease. Ovarian remnants also may be included within pelvic peritoneal inclusion cysts. In this retrospective study, the sonographic features of ovarian remnants in 10 patients with surgical proof or clinical follow-up data are described. Most ovarian remnants were simple cysts (seven of 10), three had multiple septations, and six had a rim of presumably ovarian tissue with arterial and venous flow. Three patients with ovarian remnant masses that were aspirated had symptomatic relief without recurrence. In one patient, guided aspiration was unsuccessful, probably owing to the presence of organized hemorrhage within the mass. Extrinsic compression of the distal ureter was observed in one patient, who was treated with gonadotropin releasing hormone agonist (Lupron). The sonographic findings of a completely cystic or multiseptated pelvic mass with a rim of vascularized solid tissue in a postoophorectomy patient, although such cases are rare, suggest the diagnosis of an ovarian remnant. If the diagnosis can be established with a high degree of certainty, sonographically guided aspiration may be attempted in an effort to provide symptomatic relief. Otherwise, sonography is useful in serial assessment of these masses in patients receiving medical treatment. PMID- 9733173 TI - Hill-Sachs lesion in recurrent shoulder dislocation: sonographic detection. AB - In a prospective study 61 patients with recurrent anterior shoulder dislocation were evaluated by sonography, radiography, and surgery to determine the value of sonography in the detection of a HillSachs lesion. The group consisted of 57 male and four female patients with an average age of 27 years. Hill-Sachs lesion was found in 54 (88%) shoulders of the 61 surgically treated patients. Using surgical findings as the gold standard, we found sonography to be 96% (52 of 54 cases) sensitive, 100% specific (seven of seven cases), and 97% (59 of 61 cases) accurate in the diagnosis of HillSachs lesion. The average size of the lesion measured by sonography was 19.2 mm long, 16.0 mm wide, and 4.1 mm deep. The lesion was of small or medium size (up to 6 mm deep) in 88% of patients. Results of our study show that sonography is a valuable imaging technique in the diagnosis of Hill-Sachs lesion. It produced only two false-negative results when compared with surgical findings. PMID- 9733174 TI - Transcranial Doppler ultrasonography to evaluate need for cerebrospinal fluid drainage in hydrocephalic children. AB - The aim of this study was to determine whether the resistive index in the anterior cerebral artery, as measured by transcranial Doppler ultrasonography without and with pressure provocation, predicts the need for cerebrospinal fluid drainage in hydrocephalic children. Both without and with pressure provocation, the resistive index was significantly higher (P > 0.05) in patients with raised intracranial pressure compared with control group patients and dropped significantly after drainage. With receiver operating characteristic analysis, the optimal cutoff point between normal and abnormal resistive index values was determined at 0.71 without pressure provocation and at 0.90 with pressure provocation. The addition of the pressure provocation test improved accuracy from 81 to 91%, mainly by improving specificity. In conclusion, transcranial Doppler ultrasonography with pressure provocation accurately identifies hydrocephalic children who require cerebrospinal fluid drainage procedures. PMID- 9733175 TI - Intraoperative ultrasonographically guided cryoablation of renal masses: initial experience. AB - The purpose of this study was to evaluate the feasibility of intraoperative ultrasonography to guide cryoablation of renal masses. Renal cryoablation was performed on six patients with solid renal tumors. Under ultrasonographic guidance, cryoprobes measuring 3 mm in diameter were placed into the renal tumor parenchyma or into surrounding normal parenchyma. Intraoperative ultrasonography accurately delineated tumor size, cryoprobe placement, and depth of freezing. An echogenic interface was generated by the marked impedance differences at the junction of the normal renal parenchyma and frozen tissue. In addition, intraoperative ultrasonography identified a total of nine additional lesions in three patients that were not detected by preoperative imaging. These lesions also were treated cryosurgically during the same operation. There were no deaths. The patients have been followed with clinical and laboratory assessments as well as with MR imaging or CT scanning, and all have remained tumor free 3 to 22 months postoperatively. Ultrasonographically guided renal cryoablation is a feasible technique for treating malignant renal tumors while preserving renal parenchyma. Long-term follow-up studies in a larger series of patients are required to determine the true efficacy and safety of this procedure. PMID- 9733176 TI - Femoral vein pseudoaneurysm: uncommon complication of femoral vein puncture. PMID- 9733177 TI - Focal peliosis hepatis resembling metastatic liver tumor. PMID- 9733178 TI - Fryns syndrome: prenatal diagnosis and pathologic correlation. PMID- 9733179 TI - Echogenic fetal bowel and calcified meconium in a fetus with trisomy 21. PMID- 9733180 TI - Frequent occurrence of underreporting multiple gestations based on vaginal and abdominal ultrasonograms performed prior to 6 weeks' gestational age. PMID- 9733181 TI - Otocephaly: prenatal sonographic diagnosis. PMID- 9733182 TI - The diagnosis of Spigelian hernia (SH) by high-resolution real-time sonography. PMID- 9733183 TI - Performance of the basic fetal cardiac ultrasound examination. PMID- 9733184 TI - Cue control and head direction cells. AB - Previous research has shown that head direction (HD) cells in both the anterior dorsal thalamus (ADN) and the postsubiculum (PoS) in rats discharge in relation to familiar, visual landmarks in the environment. This study assessed whether PoS and ADN HD cells would be similarly responsive to nonvisual or unfamiliar environmental cues. After visual input was eliminated by blindfolding the rats, HD cells maintained direction-specific discharge, but their preferred firing directions became less stable. In addition, rotations of the behavioral apparatus indicated that some nonvisual cues (presumably tactile, olfactory, or both) exerted above chance stimulus control over a cell's preferred firing direction. However, a prominent auditory cue was not effective in exerting stimulus control over a cell's preferred direction. HD cell activity also was assessed after rotation of a novel visual cue exposed to the rat for 1, 3, or 8 min. An 8-min exposure was enough time for a novel visual cue to gain control over a cell's preferred direction, whereas an exposure of 1 or 3 min led to control in only about half the sessions. These latter results indicate that HD cells rely on a rapid learning mechanism to develop associations with landmark cues. PMID- 9733185 TI - The hippocampus and transverse patterning guided by olfactory cues. AB - Normal rats and rats with hippocampal system damage were trained on a novel, olfactory version of the transverse-patterning task that involved the concurrent learning and continued performance of 3 partially ambiguous discrimination problems (A+B-, B+C-, C+A-). Animals with lesions of the fornix or perirhinal entorhinal cortex acquired at least as rapidly as normal rats these problems presented in sequential blocks of trials involving the same stimulus pair. All groups also performed well on an initial test session when the order of stimulus pair presentations was randomized. Normal rats continued to discriminate appropriately in additional testing sessions with trials presented in random order. By contrast, both groups with hippocampal system damage performed poorly in continued random-order testing. These results extend the generality of the deficit in transverse patterning to the olfactory modality and demonstrate that the deficit is equivalent in magnitude after fornix or perirhinal-entorhinal damage. Findings also suggest that the transverse-patterning problem can be acquired transiently without critical hippocampal involvement, although continued performance relies on hippocampal function. PMID- 9733186 TI - Lesions of the frontal cortex, hippocampus, and intralaminar thalamic nuclei have distinct effects on remembering in rats. AB - Lesions of the intralaminar thalamic nuclei (ILn), the medial wall (MW) area of prefrontal cortex, and the hippocampus were compared and found to have distinct effects on delayed matching-to-sample (DMS) and delayed non-matching-to-sample (DNMS) tasks based on different types of stimulus cues. Hippocampal lesions impaired DNMS trained in a radial arm maze but had little effect on DMS trained with retractable levers or olfactory DNMS. MW lesions affected the DMS task but had limited effects on olfactory DNMS and radial arm maze DNMS. ILn lesions resulted in a more generalized pattern of impairment for radial maze tasks and (in previous studies) for the DMS and olfactory DNMS tasks. Only the hippocampal lesion was associated with a delay-dependent impairment. It is argued that ILn lesions disrupt remembering through their effects on the recurrent, feedback pathways that link functionally related areas of the basal ganglia and cortex. PMID- 9733187 TI - Intensity discrimination and auditory brainstem responses in cochlear implant and normal-hearing listeners. AB - Intensity-discrimination limens (IDLs) and auditory brainstem responses (ABRs) were measured as a function of stimulus intensity in 6 cochlear implant (CI) and 8 normal-hearing (NH) listeners. Pulse-train stimuli were delivered electrically to the auditory nerve in CI listeners and acoustically in NH listeners. In CI listeners, the IDLs expressed as Weber fractions decreased monotonically with increasing intensity. In NH listeners, a nonmonotonic IDL function showing a peak a midintensities was observed. ABR wave amplitudes increased regularly with intensity only in CI listeners. Results support the notion that the slight decrease in Weber's fractions with increasing sound intensity--generally referred to as "the near-miss to Weber's law"--is subtended by retrocochlear processes, whereas the increase in Weber's fractions at midlevels--known as "the severe departure from Weber's law"--originates in cochlear mechanisms. PMID- 9733188 TI - Dissociation of visual and auditory pattern discrimination functions within the cat's temporal cortex. AB - In ablation-behavior experiments performed in adult cats, a double dissociation was demonstrated between ventral posterior suprasylvian cortex (vPS) and temporo insular cortex (TI) lesions on complex visual and auditory tasks. Lesions of the vPS cortex resulted in deficits at visual pattern discrimination, but not at a difficult auditory discrimination. By contrast, TI lesions resulted in profound deficits at discriminating complex sounds, but not at discriminating visual patterns. This pattern of dissociation of deficits in cats parallels the dissociation of deficits after inferior temporal versus superior temporal lesions in monkeys and humans. PMID- 9733189 TI - Contributions of the hippocampus, amygdala, and dorsal striatum to the response elicited by reward reduction. AB - Rats were trained to run down a runway for either 1 or 10 food pellets. After training, those receiving 10 pellets were shifted to 1 pellet. Such shifts typically elicit a temporary decrease in running speed. Groups of normal rats and rats with bilateral lesions of the fimbria-fornix, lateral-basolateral complex of the amygdala, or dorsal striatum were tested with the shifted and unshifted procedures. Separate experiments, identical except for the intertrial intervals (ITIs; 3 min vs. 30 s), were carried out. The data are consistent with the view that an integrated action of multiple neural systems is required to observe the typical response to reward reduction in unlesioned rats. One system that includes the dorsal striatum promotes a reinforced approach response to the goal box. A neural system that includes fimbria-fornix is required to retain information about reduced reward over the 3-min ITI. A system that includes the amygdala may acquire a conditioned aversive response to the goal box after the shift is detected, leading to reduced speeds over testing. PMID- 9733190 TI - Memory impairment on a delayed non-matching-to-position task after lesions of the perirhinal cortex in the rat. AB - Previous research conducted in monkeys and rats has established that the perirhinal cortex is critically involved in object- or stimulus-recognition memory, whereas other research suggests this region may contribute to memory for object discriminations. These findings do not rule out the possibility that the perirhinal cortex plays a more general role in memory. The present experiment addressed whether selective lesions of the perirhinal cortex would result in a delay-dependent deficit on a test of memory that did not involve stimulus recognition or object memory. Rats with bilateral perirhinal lesions were tested on a delayed non-matching-to-position task. Lesions of the perirhinal cortex did not interfere with acquisition or performance at short (0-4 s)-delay intervals, but lesions did impair performance at longer delays. It is suggested that the perirhinal cortex is involved in maintaining representations of trial-specific information over time. PMID- 9733191 TI - Neuronal plasticity induced by fear conditioning is expressed during paradoxical sleep: evidence from simultaneous recordings in the lateral amygdala and the medial geniculate in rats. AB - The lateral amygdala (LA) and its afferent connections from the medial geniculate (MG) play a pivotal role in auditory fear conditioning. The authors evaluated whether those neurons could express in paradoxical sleep (PS) physiological plasticity acquired in waking. After a habituation session, rats received tone footshock pairings in 3 sessions. After each session, the tone alone was presented during PS episodes. Multiunit activity was simultaneously recorded in the LA and the medial part of the MG. Both in LA and MG, conditioned responses emerged rapidly (within 5 trials), were expressed with short latency (<20 ms), and were maintained in PS after training. Such changes were not observed in pseudoconditioned rats. These results are discussed regarding the question of the primary sites of plasticity in auditory fear conditioning and regarding the functional significance of preserved expression in PS of learning-induced neuronal plasticity. PMID- 9733192 TI - The dorsal hippocampus is essential for context discrimination but not for contextual conditioning. AB - The authors describe how (a) the timing of hippocampal lesions and (b) the behavioral-representational demands of the task affect the requirement for the hippocampus in contextual fear conditioning. Post- but not pretraining lesions of the hippocampus greatly reduced contextual fear conditioning. In contrast, pretraining lesions of the hippocampus abolished context discrimination, a procedure in which mice are trained to discriminate between 2 similar chambers (shock context vs. no-shock context). Whereas either contextual- or cue-based strategies can be used to recognize an aversive context, discrimination between similar contexts is optimally acquired by contextual (hippocampal)-based strategies. In keeping with the lesion results, Nf1(+/-)/Nmdar1(+/-) mutant mice, which have spatial learning deficits, are impaired in context discrimination but not in contextual conditioning. Together, these data dissociate hippocampal and nonhippocampal contributions to contextual conditioning, and they provide direct evidence that the hippocampus plays an essential role in the processing of contextual stimuli. PMID- 9733193 TI - Opposite effects of lateral septal LTP and lateral septal lesions on contextual fear conditioning in mice. AB - The effect of fimbrial high-frequency stimulation (HFS)-induced long-term potentiation (LTP) in the lateral septum (LS) on contextual fear conditioning was studied in mice. Mice were conditioned for fear toward a novel context through the use of footshocks. The 1st experiment showed that pretraining HFS reduced significantly conditional freezing to contextual stimuli. The 2nd experiment was designed to determine whether the reduction of freezing produced by fimbrial HFS resulted from LTP in the LS rather than from LTP in other brain structures. Accordingly, mice with lesions of the LS were used and submitted to the same protocol as in the 1st experiment. Results showed that LS lesions completely abolished the impairing effect of fimbrial HFS and, as a whole, potentiated the freezing response. These data suggest that contextual fear conditioning is strongly modulated by the level of hippocampal-LS synaptic neurotransmission. PMID- 9733194 TI - Correlational relationship between shock intensity and corticosterone secretion on the establishment and subsequent expression of contextual fear conditioning. AB - A role for corticosterone in the consolidation of contextual fear conditioning has previously been proposed. In this study, physiological evidence was found to support this view. The extent of conditioned fear and the levels of plasma corticosterone in rats, after context exposure at training and at different posttraining times (24 hr and 7 days), depended on the intensity of the unconditional stimulus (footshock). In each experimental session, a positive correlation was found between the magnitude of corticosterone levels and the fear related behavioral inhibition exhibited in the context. Results support the involvement of corticosterone on the processes that occur during consolidation in determining the strength at which the contextual fear conditioning is stored as a long-term memory. PMID- 9733195 TI - Comparison of the amnestic effects of NMDA receptor antagonist MK-801 and nitric oxide synthase inhibitors: L-NAME and L-NOARG in goldfish. AB - Investigations indicate that the induction of long-term potentiation (LTP) may be mediated by postsynaptic N-methyl-D-aspartate (NMDA) receptors and that the maintenance of LTP may be initiated by nitric oxide (NO), a retrograde messenger carrying signals backward from the postsynaptic to the presynaptic neuron. The present study compared amnestic effects of dizocilpine maleate (MK-801), an NMDA receptor antagonist, and nitro-L-arginine-methyl-ester (L-NAME) and N-nitro-L arginine (L-NOARG), nitric oxide (NO) inhibitors, in goldfish, using active avoidance conditioning as the learning paradigm. The results showed that MK-801 and NO inhibitors produced anterograde amnesia at doses that did not impair performance processes necessary for learning to occur. Furthermore, MK-801 did not produce retrograde amnesia, whereas L-NAME did, suggesting that MK-801 impaired learning whereas NO inhibitors impaired memory consolidation and possibly also learning. PMID- 9733196 TI - The opposite effects of cysteamine on the acquisition of two different tasks in mice are associated with bidirectional testing-induced changes in hippocampal adenylyl cyclase activity. AB - The hypothesis of a role for hippocampal somatostatin (SS-14) in learning and memory processes was further examined by means of 2 selective learning tasks that were previously shown to be either impaired (spatial discrimination task) or facilitated (barpressing task) by hippocampal lesions. Results showed that subcutaneous injections of cysteamine (160 mg/kg) (a) impaired acquisition of the spatial task while producing an opposite (i.e., facilitative) effect on acquisition of the barpressing task and (b) produced an up regulation of hippocampal adenylyl cyclase (AC) activity, which was antagonized by spatial discrimination training but enhanced by training in the barpressing task. Moreover, opposite task-dependent training-induced changes in hippocampal AC activity was observed in saline-treated mice. These results suggest that bidirectional regulatory mechanisms of hippocampal function involving both SS-14 and ACs may occur as a function of the type of learning. PMID- 9733198 TI - Ontogenetic differences in the expression of conditioned stimulus conditioning: effects of retention interval. AB - Preweanling 17-day-old rats were tested for retention of the conditioned emotional response after a 5-min or 24-hr retention interval. For a variety of conditioning parameters (i.e., variation in conditioned stimulus modality, unconditioned stimulus intensity, number of training trials), conditioned responding was consistently weaker after 5 min than after 24 hr. This apparent "incubation," or "hypermnesic," effect was not found in adult rats, even when comparable conditioning levels were indicated on the 24-hr test. The transient short-term retention deficit observed in 17-day-old preweanlings was alleviated by placing the pup in its home cage during the 5-min retention interval or by extending the conditioning session. Fifteen-day-old rat pups did not benefit from home cage exposure or extended training and displayed the transient short-term retention deficit regardless. The results are discussed in terms of age-related effects on time-dependent memory consolidation. PMID- 9733197 TI - Reactivation treatment prevents the memory-impairing effects of scopolamine in preweanling rats. AB - The authors report that the expression of a conditioned odor aversion is impaired in preweanling rats when they are conditioned on Postnatal Day 12 and tested under the influence of scopolamine hydrobromide (0.2 or 0.5 mg/kg, intraperitoneal) after a 48-hr, but not after a 2-hr, retention interval (Experiment 1). This effect of scopolamine is not dependent on maturation of the cholinergic system between Days 12 and 14 (Experiment 2), nor is it due to peripheral mechanisms (Experiment 3). When pups are reexposed to the unconditioned stimulus (footshock) before drug administration, performance on the 48-hr retention test is not impaired by scopolamine (Experiment 4). These findings demonstrate that the cholinergic system may be critical for the retrieval and expression of long-term or weak memories in young rats. However, the expression of active memories (recent or recently reactivated) may not be dependent on the cholinergic system to the same extent as is the expression of inactive memories. PMID- 9733199 TI - Isolation-stress-induced facilitation of passive avoidance memory in the day-old chick. AB - This study showed that facilitation of recall of a weak version of the 1-trial passive avoidance learning task could be achieved by behavioral "stressing" of day-old chicks after training. Recall, usually retained for less than 9 hr, was extended by socially isolating the chicks for 1 hr immediately after training. There was a brief 3-fold increase in plasma corticosterone levels 10 min after isolation. Facilitated recall was not evident when chicks were isolated 2 hr after training, and it was blocked by intracerebral administration of 2-ng RU 38486, a specific glucocorticoid receptor antagonist, per chick. Male chicks responded more to isolation than did female chicks, presumably a consequence of the additional stress of the injection procedure. PMID- 9733200 TI - Aging and ischemia in gerbils impair spatial memory performance. AB - Gerbils aged 3 months and 24+ months were subjected to 5 min of global forebrain ischemia and tested in a radial arm maze (1 trial/day, 50 days). Compared with age-matched, sham-operated controls, ischemic animals were impaired on measures of both working and reference memory. Aged animals were impaired on working memory, but not on reference memory, compared with their younger counterparts. Hippocampal CA1 and CA2 regions were significantly and comparably damaged in the 2 ischemic groups but were unaffected by aging. The results suggest that aging and ischemia have functionally similar effects on working memory, but the 2 processes differentially impact reference memory. PMID- 9733201 TI - Social isolation blocks the expression of memory after training that a food is inedible in Aplysia fasciata. AB - Isolating a sexually mature Aplysia fasciata for either 1 or 24 hr immediately after training that a food is inedible blocks the subsequent expression of memory measured 24 hr later. Isolation that is delayed for 1 hr after training, but not for 12 hr after training, is also effective in blocking memory. Isolation affects memory because of a specific effect caused by the absence of pheromones secreted by conspecifics rather than by a nonspecific change in the chemical environment, because transferring animals to a novel environment (120% seawater) that contains a conspecific does not affect memory. Isolation also does not affect memory in sexually immature Aplysia, even though immature animals are able to sense one another's presence. Isolation may affect memory because social (and sexual) isolation is a form of stress in mature A. fasciata, and stress after training affects retention in many animals. PMID- 9733202 TI - The effects of cocaine, amphetamine, and the dopamine D1 receptor agonist SKF 38393 on fear extinction as measured with potentiated startle: implications for psychomotor stimulant psychosis. AB - Using Pavlovian conditioned increases in the amplitude of the acoustic startle reflex as a behavioral indicator of fear motivation, the authors previously showed a resistance to extinction after repeated associations of cocaine with the fear-evoking conditioned stimulus (CS). In Experiment 1, acute administration of cocaine, amphetamine, and the dopamine (DA) D1 receptor agonist SKF 38393 produced a similar fear enhancement. In Experiment 2, a noncontingent injection of cocaine and SKF 38393 provoked a CS potentiation of acoustic startle in fear extinguished laboratory rats. Potential behavioral, neurochemical, and neuroendocrine explanations for the effects of psychomotor stimulants on conditional fear were discussed. It was suggested that DA agonist drugs increase fear expression possibly by activating mesoamygdaloid associative neurocircuitry involved in excitatory conditioned fear reactions. PMID- 9733203 TI - Effects of systemic, intracerebral, or intrathecal administration of an N-methyl D-aspartate receptor antagonist on associative morphine analgesic tolerance and hyperalgesia in rats. AB - A flavor paired with morphine shifted to the right the function relating morphine dose to tail-flick latencies and provoked hyperalgesic responses when rats were tested in the absence of morphine. These learned increases in nociceptive sensitivity were not mediated by alterations in tail-skin temperature. Microinjection of the competitive N-methyl-D-aspartate (NMDA) receptor antagonist D,L-2-amino-5-phosphonopentanoic acid (AP-5) into the lateral ventricle reversed the hyperalgesic responses but spared the tolerance to morphine analgesia. By contrast, systemic administration of the noncompetitive NMDA receptor antagonist MK-801 or intrathecal infusion of AP-5 reversed the hyperalgesic responses as well as the tolerance to morphine analgesia. The results demonstrate that associatively mediated tolerance to morphine analgesia can co-occur with hyperalgesic responses and are discussed relative to learned activation of endogenous pronociceptive mechanisms. PMID- 9733204 TI - Laterally asymmetrical cell number in a sexually dimorphic nucleus in the gerbil hypothalamus is correlated with vocal emission rates. AB - A lateralized relationship between the total volume of a discrete area in a hypothalamic, sexually differentiated nucleus (SDApc) and stereotyped vocalization exists in male Mongolian gerbils. In this present study, using unbiased stereological methods, two cytoarchitectural estimates of the SDApc's structure, neuron number, and nuclear volume were found to be sexually differentiated and also laterally asymmetrical in adult males. In ovariectomized females receiving exogenous testosterone, no cytoarchitectural component was asymmetrical. Significantly, the estimate of neuron number, but not nuclear volume, in the left SDApc of males was correlated with vocal emission rate. The authors conclude that a specific, sex-related cytoarchitectural SDApc parameter shows left-right asymmetry, suggesting the SDApc has an intimate role in mediating hemispheric specialization rather than just being an end point index of individual structural variability. PMID- 9733205 TI - Taste responses in the greater superficial petrosal nerve: substantial sodium salt and amiloride sensitivities demonstrated in two rat strains. AB - A great quantity of research has focused on neural responses of the chorda tympani nerve (CT) to taste stimuli. This report examined salt and sugar sensitivity of the greater superficial petrosal nerve (GSP) and the effect of amiloride on these neural responses. In addition to Sprague-Dawley (SD) rats that have CT responses typical of most rat strains, we included Fischer 344 (F344) rats whose CT responses to sodium chloride (NaCl) are higher than those of other strains. After a stimulation series in which water served as the rinse, a series of stimuli was presented in 100 microM amiloride. The GSP was highly responsive to NaCl, sodium acetate (NaAc), ammonium chloride, and sucrose; NaCl and NaAc responses were strongly suppressed by amiloride. Relative responses to NaCl were significantly higher in F344 than in SD rats. In summary, the GSP is highly sensitive to salt and sugar stimulation, and palatal taste receptors have a considerable degree of amiloride sensitivity. PMID- 9733206 TI - Analgesic responses of male mice exposed to the odors of parasitized females: effects of male sexual experience and infection status. AB - The present study shows that parasites influence both the responses of males to infected females and the responses of male hosts to females. Male mice exposed for 30 min to the odors of females infected with the nematode parasite Heligmosomoides polygyrus displayed a naloxone-sensitive, opioid-mediated analgesia, whereas males exposed for 1 min showed a shorter duration and lower amplitude naloxone-insensitive "nonopioid" analgesia that involved serotoninergic (5-HT) and excitatory amino acid (N-methyl-D-aspartate [NMDA] receptor) systems. The male mice distinguished between the odors of infected and physically stressed females, displaying greater analgesia after exposure to the odors of infected than stressed females. The analgesic responses to the odors of infected females were also affected by the males' prior sexual experience; sexually experienced males exhibited significantly greater analgesia than sexually naive males. In contrast, male mice infected with H. polygyrus failed to show a nonopioid analgesia after exposure to the odors of infected females and displayed a markedly lower level of opioid analgesia than uninfected mice. These results show that male mice can discriminate between the odors of parasitized and nonparasitized females and find the odors of parasitized estrous females aversive. PMID- 9733207 TI - Learning upregulates brain-derived neurotrophic factor messenger ribonucleic acid: a mechanism to facilitate encoding and circuit maintenance? AB - Brain-derived neurotrophic factor (BDNF) promotes neuron survival, enhances sprouting, protects neurons against insult, and may be involved in several aspects of learning and memory. In this study, rats trained to locate a submerged platform in a water maze had elevated levels of BDNF messenger ribonucleic acid (mRNA) in the hippocampus (p < .05), a structure associated with spatial memory. BDNF mRNA expression increased after 3 and 6 days but not after 1 day of training in the water maze. A yoked control group that swam without the platform present, to control for physical activity, showed a trend for elevated BDNF mRNA at an intermediate level between the learning and sedentary groups. Other cortical and subcortical areas did not show a significant increase in BDNF mRNA after learning or activity (p > .05). These findings suggest that learning can impact BDNF mRNA expression localized to the brain areas involved in the processing of spatial information. Furthermore, behaviors such as physical activity and learning may help maintain and protect neurons at risk in aging and neurodegenerative disease via increased BDNF expression. PMID- 9733209 TI - Effects of gonadectomy in infancy and adulthood on handedness in male and female Mongolian gerbils. AB - When assuming a species-typical tripodal posture, female Mongolian gerbils most often rest on their left forepaws and hold their right forelimbs aloft; male gerbils most often do the reverse. This experiment examined effects of gonadectomy, both in infancy and in adulthood, on the sexually dimorphic asymmetry in forelimb use by Mongolian gerbils when maintaining a tripodal stance. In adulthood, both male and female gerbils that had been gonadectomized at birth reversed their forelimb use while in a tripodal stance: Gonadectomized males used their forelimbs as did sham-operated females, and gonadectomized females used their forelimbs as did sham-operated males. Gonadectomy in adulthood abolished the sexual dimorphism in forelimb use seen in sham-operated subjects. It was concluded that gonadal hormones have organizational as well as possible activational effects on adult patterns of forelimb use by gerbils. PMID- 9733208 TI - Rhinal cortex ablations fail to disrupt reinforcer devaluation effects in rhesus monkeys (Macaca mulatta). AB - Studies have shown that excitotoxic lesions of the amygdala attenuate reinforcer devaluation effects in monkeys and rats. Because the rhinal (i.e., entorhinal and perirhinal) cortex has prominent reciprocal connections with the amygdala and has been suggested to store knowledge about objects, it is possible that it too composes part of the critical circuitry subserving learning about objects and their associated reinforcement value. To test this possibility, rhesus monkeys with rhinal cortex removals as well as unoperated controls were tested using a reinforcer devaluation procedure. Monkeys with rhinal cortex removals and controls, unlike those with amygdala lesions, tended to avoid displacing objects overlying a devalued food. These results indicate that the rhinal cortex is not a critical part of the neural circuitry mediating the effects of reinforcer devaluation. PMID- 9733211 TI - Lack of predictability of classical animal models for hypolipidemic activity: a good time for mice? AB - Hypolipidemic drugs that are efficacious in man are not always active in classical animal models of dyslipidemia. Inhibitors of HMG-CoA reductase (statins) do not lower plasma cholesterol in rats, but yet this species was alone in providing activity for fibrate-type drugs. Nicotinic acid possesses many desirable features with regard to clinical use, but most of these actions are lacking in rats and monkeys. The metabolism of low density lipoproteins in hamsters is widely thought to be similar to that in humans, yet neither statins or fibrates lower plasma lipids in these species. With the advent of mouse models expressing specific human genes (or disruption of genes) it is now possible to re examine the effect of established drugs and to characterize new hypolipidemic compounds with respect to site and mechanism of action. Drug responses observed in humans are now being seen in such mouse models (e.g. HDL elevation with fenofibrate in mice with the human apo A-I gene). Moreover, mice are now being screened for compounds that lower plasma (human) Lp(a), or lower plasma cholesterol in the absence of LDL receptors. It is proposed that these new genetic mouse models may afford a more focused examination of drug action and provide, for new compounds, better prediction of the human response. PMID- 9733210 TI - Nitric oxide and endothelin in the development of cardiac allograft vasculopathy. Potential targets for therapeutic interventions. AB - Extensive research has been carried out in recent years to discover the potential risk factors contributing to cardiac allograft atherogenesis. Injury to endothelial cells has been regarded as an important early mechanism in the development of transplant atherosclerosis; it leads to the manifestation of epicardial and microvascular endothelial dysfunction and development of intimal hyperplasia. Moreover, continuous minor endothelial cell damage contributes to endothelial dysfunction which reflects one of the first measurable steps in the cascade of atherogenesis without macroscopic evidence of vascular lesions. The discovery of two important vasoactive substances nitric oxide (NO) and endothelin (ET) has brought new insights but also new unsolved questions regarding the mechanisms leading to atherosclerosis. To date it is known that both substances play a major role in both prevention and development of atherosclerosis. NO appears to be protective in low concentrations by inhibiting leukocyte and platelet activation/adherence and smooth muscle cell proliferation. Impaired endothelial NO production, as one cause of endothelial dysfunction may occur in early stages of atherosclerosis before macroscopic lesions are evident. In addition, increased endothelin release also results in endothelial dysfunction by inducing vasoconstriction; it promotes vascular lesion formation due to endothelial- and vascular smooth muscle cell proliferation. Direct and indirect manipulation of both the NO and ET signal transduction systems may provide novel preventive and therapeutic approaches for limiting transplant atherogenesis and to treat native atherosclerosis. This review summarizes important experimental and clinical evidence which points to nitric oxide and endothelin as potential therapeutic targets in the process of cardiac allograft vasculopathy. PMID- 9733212 TI - Transforming growth factor-beta 1 and ascorbate regulate proliferation of cultured smooth muscle cells by independent mechanisms. AB - We previously reported that ascorbate (vitamin C) can regulate the growth of cultured vascular smooth muscle cells (VSMC) directly as well as by altering the properties of extracellular matrix (ECM) [Mol Cell Cardiol 1997;29:3293-303]. In the present study we compared the effects of ascorbate and transforming growth factor-beta 1 (TGF-beta1) on VSMC growth in order to determine whether their actions were mediated by similar mechanisms. When VSMC proliferation was stimulated by fetal bovine serum, the addition of TGF-beta1 (20 ng/ml) or ascorbate (1 mM) to the cell culture medium inhibited the cellular incorporation of [3H]thymidine by 19 and 59%, respectively, and by 85% when added together. The cell growth inhibitory effects of TGF-beta1 and ascorbate were partially mediated by changing the growth-regulatory properties of the ECM produced by the cells. Thus, VSMC grew more slowly on ECM deposited by VSMC under treatment with 20 ng/ml TGF-beta1 or 1 mM ascorbate (52 and 46% inhibition, respectively) than on control ECM, and their combination had an additional inhibitory effect (84%). Anti-TGF-beta1 neutralizing antibodies prevented the direct and ECM-mediated effects of TGF-beta1 on VSMC growth, but did not alter the effects of ascorbate. When ECM was pre-incubated with increasing concentrations of TGF-beta1, the growth rate of freshly plated VSMC gradually decreased, indicating that ECM-bound TGF-beta1 retained its biological activity. Comparison of the patterns of TGF betal binding to ECM produced by VSMC in the presence or absence of ascorbate revealed no significant differences. Extraction of ECM-bound TGF-beta1 by incubation of exposed ECM with plasmin did not affect the ECM-mediated inhibitory effect of ascorbate, as the rate of proliferation of secondary VSMC cultures grown on ascorbate-dependent and independent matrices treated with plasmin were equally increased. These results suggest that the amount of ECM-bound TGF-beta1 was not altered by ascorbate. The secretion of TGF-beta1 into the cell culture medium by VSMC also did not depend on the ascorbate supply. Finally, addition of heparin to the VSMC culture medium during ECM production abolished the ECM mediated growth inhibitory effects of ascorbate, but did not affect the action of TGF-beta1. Our data demonstrate that the growth inhibitory effects of ascorbate on cultured VSMC are independent of the action of TGF-beta1, and the effects of these two compounds on VSMC growth are additive. PMID- 9733213 TI - Cholesteryl esterase-treated LDL augments oxidized LDL-mediated cholesteryl ester deposition in mouse peritoneal macrophages. AB - Arterial unesterified cholesterol, phospholipid particles have been isolated from atherosclerotic lesions and characterized. However, the role of these 'liposomes' in macrophage foam cell formation is unclear. Recently, LDL, after trypsin and cholesteryl esterase treatment (T/CE LDL), was shown to have physical properties similar to the unesterified cholesterol, phospholipid particles isolated from atherosclerotic lesions. Yet, when mouse peritoneal macrophages were incubated with these model particles in culture medium (DMEM and 5% LPDS), only an insignificant accumulation of cellular cholesteryl esters was observed. Previously, we demonstrated that complex formation between unesterified cholesterol, phosphatidylcholine liposomes and cupric sulfate-oxidized LDL dramatically enhances the ability of the liposomes to augment cellular cholesterol accretion (Greenspan P, Yu H, Mao F, Gutman RL. J Lipid Res 1997;38:101-109). When T/CE LDL, another cholesterol-rich phospholipid particle, was substituted for unesterified cholesterol phosphatidylcholine liposomes in our complex, mouse peritoneal macrophages accumulated a significant amount of both cellular unesterifed cholesterol (61 microg/mg cell protein) and cholesteryl esters (76 microg/mg cell protein) after 48 h of incubation. These results demonstrate again that the interaction of two cholesterol-bearing particles (T/CE LDL and oxidized LDL), which individually can not promote significant cholesterol accumulation in cells, will, when combined, produce macrophage foam cells. PMID- 9733214 TI - Increased PAI activity and PAI-1 antigen occurring with an oral fat load: associations with PAI-1 genotype and plasma active TGF-beta levels. AB - Whether the post-prandial lipemic response is linked to potentially pro atherogenic and/or prothrombotic changes in plasminogen activator inhibitor (PAI) and transforming growth factor-beta (TGF-beta) is uncertain. The aim of our study was to determine whether PAI-1 antigen and PAI activity were elevated during post prandial lipemia following a standard fat tolerance test. We also investigated changes in TGF-beta1 antigen and TGF-beta activity, to determine whether changes in TGF-beta activity were associated with changes in PAI measurements. Lastly, the influence of genotype at a common insertion/deletion polymorphism in the PAI 1 promoter on changes in PAI activity and PAI-1 antigen was examined. Fat tolerance tests were undertaken in 57 healthy middle-aged men to investigate associations between plasma concentrations of lipoproteins, PAI (antigen and activity) and TGF-beta. PAI-1 concentration increased by 76% after 8 h (P < 0.0001). PAI activity also increased by 64% (P = 0.0054) and TGF-beta activity decreased by 10% (P < 0.0001). Increases in PAI-I antigen and PAI activity varied markedly between individuals. To investigate these heterogeneous responses we examined whether genotype at the common insertion/deletion polymorphism of the PAI-1 promoter accounted for these differences. Individuals with at least one 4G (deletion) allele showed potentially pro-atherogenic changes in both PAI-1 and TGF-beta, compared to individuals who were homozygous for the 5G (insertion) allele. In conclusion, increased PAI and decreased TGF-beta activity occur during a fat tolerance test and this effect may be modulated by a common insertion/deletion polymorphism in the PAI-1 promoter. PMID- 9733215 TI - Relationships between protein C, protein S, von Willebrand factor and euglobulin lysis time and cardiovascular risk factors in subjects with and without coronary heart disease. AB - Measures of fibrinolytic and thrombotic function have been examined in 55 subjects with recently identified coronary heart disease, and age and sex matched control subjects. Measurements were particularly directed at factors and processes which could be affected by changes in endothelial function and included the euglobulin lysis time as well as plasma levels of von Willebrand factor (vWF). Plasma levels of protein S and protein C were also measured. Measurements were made before and after a period of 10-min veno-occlusion combined with rhythmic hand exercise. In addition anthropometric, haemodynamic and biochemical measurements (plasma lipids and apolipoproteins, glucose and insulin) were obtained and correlated with the haematological parameters. Protein S and vWF levels were significantly higher, both before and after veno-occlusive exercise, in subjects with CHD than in the asymptomatic controls. Euglobulin lysis times were not significantly different but only shortened on veno-occlusive exercise in those without CHD. Protein S levels were significantly correlated with systolic blood pressure, plasma total cholesterol, plasma triglyceride, plasma phospholipid, plasma fasting glucose and both apolipoprotein A1 and B levels. vWF levels were not significantly related to any of the other variables. Subjects whose pre-exercise euglobulin lysis times exceeded 6 h had significantly higher BMI, plasma total cholesterol, triglyceride, phospholipid, insulin, glucose and apoB concentrations and lower HDL cholesterol than those with lysis in less than 6 h. The findings from this study are consistent with a role for endothelial dysfunction in the production of atherosclerotic vascular disease and may indicate additional, non-haemodynamic, mechanisms for such an association. In addition, the relationship between elevated levels of protein S and CHD does not appear to depend on the demonstrated associations between protein S and a number of other cardiovascular risk factors. PMID- 9733216 TI - Smaller, denser LDL particles are not a risk factor for cardiovascular disease in healthy nonagenarian women of the Cremona Population Study. AB - We evaluated LDL particle size and its relation with other established risk factors for cardiovascular disease in a group of healthy nonagenarian ( > or = 90 years) women participating in the Cremona Population Study. A group of younger healthy postmenopausal women (45-75 years) was used as control group. Nonagenarian women had significantly lower body mass index, systolic and diastolic blood pressure, and fasting insulin concentrations. Plasma total, LDL and HDL cholesterol, apo AI and apo B concentrations, and LpAI and LpAI:AII particles were significantly lower in the nonagenarian group as well. LDL particle size (262.7+/-0.9 vs. 270.1+/-1.1 A) was also lower in the nonagenarian group. The presence of the E4 isoform of apo E in the nonagenarian group resulted in significantly higher levels of plasma apo AI and LpAI:AII particles, and a trend toward larger LDL particles, and a lower diastolic blood pressure. In conclusion, smaller and denser LDL particles might not represent an important risk factor for cardiovascular disease in healthy nonagenarian women of the Cremona Population Study, characterised by a reduced number of LDL particles and other protective factors, like low systolic and diastolic blood pressure, body mass index, and plasma insulin levels. PMID- 9733217 TI - Elevated plasma cholesteryl ester transfer in NIDDM: relationships with apolipoprotein B-containing lipoproteins and phospholipid transfer protein. AB - Lecithin:cholesteryl acyl transferase (LCAT) and cholesteryl ester transfer protein (CETP) are key factors in the esterification of cholesterol and the subsequent transfer of cholesteryl ester from high density lipoproteins (HDL) towards very low and low density lipoproteins (VLDL + LDL). Phospholipid transfer protein (PLTP), lipoprotein lipase (LPL) and hepatic lipase (HL) are involved in plasma phospholipid and triglyceride metabolism and also affect HDL. Equivocal changes in plasma cholesteryl ester transfer have been reported in non-insulin dependent diabetes mellitus (NIDDM). In 16 NIDDM men with plasma triglycerides < or = 4.5 mmol/l and cholesterol < or = 8.0 mmol/l. plasma cholesteryl ester transfer (CET), cholesterol esterification rate, LCAT and PLTP activity levels were higher (P < 0.05 to P < 0.02) in conjunction with higher plasma triglycerides (P < 0.01) and lower HDL cholesterol and cholesteryl ester levels (P < 0.05) compared to 16 matched healthy men. Multiple stepwise regression analysis demonstrated that CET was positively related to VLDL + LDL cholesterol (P < 0.001), triglycerides (P = 0.001), PLTP activity (P = 0.007) and CETP activity (P = 0.008, multiple r = 0.94). NIDDM had no effect on CET, independently from these parameters. HDL cholesteryl ester was negatively related to CET (P= 0.017), HL activity (P = 0.033) and NIDDM (P = 0.047) and positively to LCAT activity levels (P = 0.034, multiple r = 0.68). It is concluded that the elevated CET in plasma from NIDDM patients is associated with higher plasma triglycerides and PLTP activity levels. Furthermore, our data suggest that in normo- and moderately dyslipidaemic subjects PLTP and CETP activity levels per se may influence the rate of cholesteryl ester transfer in plasma. Plasma cholesteryl ester transfer appears to be a determinant of HDL cholesteryl ester, but other factors are likely to contribute to lower HDL cholesteryl ester levels in NIDDM. PMID- 9733218 TI - Quantitative analysis of cholesterol and cholesteryl esters in human atherosclerotic plaques using near-infrared Raman spectroscopy. AB - Raman spectroscopy is a non-destructive analytical technique and previous results have shown that qualitative analysis of the lipid component of human atheromatous arteries is feasible. In this paper, we describe a quantitative analytical method for cholesterol and cholesteryl esters in human atherosclerotic plaques, combined with Raman spectroscopic results, using partial least-squares (PLS) regression, a statistical multivariate method based on factorial analysis. Twenty-nine human atherosclerotic pooled samples were studied and the results of Raman spectroscopy coupled with the PLS method were compared to biochemical results. The standard error of prediction was 16.1, 13.6, 1.9, 3.3 and 3.4 mg/g for total cholesterol, free cholesterol, palmitate cholesteryl, oleate cholesteryl and linoleate cholesteryl, respectively. The repeatability of Raman spectroscopy was found to be excellent. Our results show that Raman spectroscopy is a promising technique to obtain a consistent and non-destructive quantitative analysis of cholesterol and cholesteryl esters in human atherosclerotic lesions. In situ and in vivo analysis is a possibility in the near future. PMID- 9733219 TI - Apolipoprotein E4, lipoprotein lipase C447 and angiotensin-I converting enzyme deletion alleles were not associated with increased wall thickness of carotid and femoral arteries in healthy subjects from the Stanislas cohort. AB - Studies have shown contrasting results concerning the relation between carotid intima-media thickness (IMT) and apolipoprotein E (apo E) and angiotensin converting enzyme (ACE) polymorphisms. Subjects, 76 men and 74 women, between 33 and 50 years, without any history of cardiovascular disease and without any anti hypertensive or lipid lowering medication were selected from the Stanislas cohort. The IMT of carotid and femoral arteries were investigated by B-mode ultrasonography. The common apo E, (C/G)447 lipoprotein lipase (LPL) and I/D ACE gene polymorphisms and serum ACE activity were determined. In the overall sample, male sex, age, systolic blood pressure, BMI, serum apo B level and tobacco consumption were positively correlated with carotid and femoral IMT. The common apo E polymorphism, the (C/G)LPL447 polymorphism and ACE activity were not related to carotid and femoral IMT variability in either men or women. Unexpectedly, the I allele of the ACE gene was related to higher femoral IMT than the D allele in non-smokers only. Similar results were observed after adjustment for the main covariates of IMT variability. In conclusion, amongst our young adult sample the candidate risk factors for cardiovascular disease, apo epsilon4, C447-LPL and D-ACE alleles and ACE activity were not associated with increased carotid and femoral IMT. PMID- 9733220 TI - Effect of estrogen on vascular smooth muscle cells is dependent upon cellular phenotype. AB - To investigate the growth-regulating action of estrogen on vascular smooth muscle cells (SMC), effects of beta-17-estradiol (beta-E2) on phenotypic modulation and proliferation of rabbit aortic SMC were observed in vitro. At 10(-8)M, beta-E2 significantly slowed the decrease in volume fraction of myofilaments (Vv myo) of freshly dispersed SMCs in primary culture, indicating an inhibitory effect of beta-E2 on spontaneous phenotypic modulation of SMC from a contractile to a synthetic phenotype. Freshly dispersed SMCs treated with beta-E2 also had a relatively longer quiescent phase than control cells before intense proliferation occurred. This was in contrast to SMCs in passage 2 3 (synthetic state), where beta-E2-treated cells replicated significantly faster than untreated cells. beta E2 also markedly enhanced the serum-induced DNA synthesis of synthetic SMCs in a concentration-dependent manner within physiological range (10(-10)to 10(-8)M). These findings indicate that the growth-regulating effect of estrogen on vascular SMC is dependent on the cell's phenotypic state. It delays the cell cycle re entry of the contractile SMCs by retarding their phenotypic modulation: however, once cells have modulated to the synthetic phenotype, it promotes their replication. PMID- 9733221 TI - The role of (E)-4-hydroxy-2-nonenal in platelet activation by low density lipoprotein and iron. AB - (E)-4-Hydroxy-2-nonenal (HNE) is a highly reactive product of the oxidation of low density lipoprotein (LDL) which increases the platelet aggregation response to various agonists. HNE formation was increased during the enhanced platelet aggregation to thrombin, ADP. A23187 and epinephrine in the presence of LDL. The increase in platelet aggregation and HNE formation by LDL was inhibited by superoxide dismutase and catalase, suggesting superoxide and hydrogen peroxide produced by platelets during aggregation may be at least partly responsible. The responsiveness of platelets to LDL and the accompanying HNE formation was increased further in the presence of ferrous ion. The effect of ferrous ion on both platelet responses and HNE formation was decreased by superoxide dismutase, catalase and the antioxidants dipyridamole and probucol implicating platelet derived free radicals. Ferrous ion caused an increase in the release of arachidonic acid from platelet membrane phospholipids in the presence of LDL which was probably caused by increased HNE production. The results suggest iron could increase platelet reactivity at sites of vascular injury by increasing HNE formation and promote the development of atherosclerotic lesions. PMID- 9733222 TI - Increased secretion of cholesteryl ester transfer protein from hamster adipose tissue: stimulation by beta-adrenergic agents. AB - High levels of cholesteryl ester transfer protein (CETP) favours decreased plasma high density lipoprotein cholesterol and increased levels of cholesterol in apolipoprotein B containing lipoproteins. Adipose tissue is one of the major sources of circulating CETP. Previous studies by our group and others demonstrated that the production of CETP from hamster adipose tissue increases after fasting, a metabolic state known to affect the sympathoadrenal axis. The present study examines the influence of beta-adrenergic agonists on the secretion of CETP from hamster adipose tissue. Fifteen minutes after an intraperitoneal injection of isoproterenol (12 microg/kg), the release of CETP mass and activity from adipose tissue fragments incubated in vitro were significantly increased. This was associated with an elevation in CETP mass and activity in plasma. The effects of isoproterenol on CETP release from adipose tissue and plasma CETP levels were suppressed by propranolol, a beta-adrenoceptor inhibitor. Addition of 10(-6) M isoproterenol to adipose tissue in vitro increased the release of CETP mass and activity from adipose tissue and this was also blocked by propranolol. Isoproterenol-induced secretion of CETP activity from adipose tissue was partially inhibited by cytochalasin B, an inhibitor of actin cytoskeleton reorganization. Forskolin, a classical adenylate cyclase agonist and 8-bromo cAMP, a functional analogue of cAMP, mimicked the effect of isoproterenol on CETP release from adipose tissue. Our results suggest that isoproterenol increases the secretion of CETP from hamster adipose tissue through a beta-adrenoceptor and a cAMP-dependent pathway. Actin cytoskeleton reorganization may be required for secretion of CETP. The findings imply that the secretion of CETP from adipose tissue is under neurosympathetic control. PMID- 9733223 TI - Association between secondary flow in models of the aorto-celiac junction and subendothelial macrophages in the normal rabbit. AB - In order to examine the association between arterial fluid dynamics and the distribution of subendothelial macrophages in the normal rabbit aorta, steady and pulsatile particle flow visualization was performed in a geometrically realistic model of the rabbit aorto-celiac junction region. Over a range of aorto-celiac steady flow ratios, particle pathlines along the upstream lateral aortic walls curved to enter the celiac orifice, while two asymmetric regions of reversing spiral secondary flow originated along the downstream lateral portions of the orifice flow divider. These regions increased in size as either the Reynolds number or flow into the celiac artery increased. In pulsatile flow studies, particles along the lateral aortic walls near the celiac orifice began to spiral into the branch during peak systole. During systolic deceleration, the size of this spiral flow region increased as particles reversed direction to enter the celiac orifice. This contrasted with flow patterns directly upstream and downstream of the orifice, which remained unidirectional throughout this period even along the distal lip of the orifice. The highest frequency of subendothelial white blood cells in the normal rabbit aorta was associated with regions where secondary flow patterns occurred, and where the orientation of endothelial cell nuclei deviated from the major direction of aortic flow. Secondary flow patterns may aid the accumulation of monocytes and macrophages about the lateral regions of the celiac artery flow divider by transporting monocytes to the walls, allowing them time to attach to the endothelial cells, or by stimulating the endothelial cells to express leukocyte adhesion molecules. These same regions are associated with increased endothelial permeability to low density lipoprotein and, under hypercholesterolemic conditions, lesion origination. PMID- 9733224 TI - Inhibition of cross-links in collagen is associated with reduced stiffness of the aorta in young rats. AB - Collagen and elastin fibres are of major importance in providing the aorta with tensile strength and elasticity. The presence of cross-links in collagen and elastin is essential for the mechanical stability of collagen and elastin fibres. beta-aminopropionitrile (BAPN) reduces the formation of cross-links by inhibiting the enzyme lysyloxidase. Young rats were injected with BAPN to inhibit the formation of cross-links, and the changes in the biomechanical and biochemical properties of the thoracic aorta were studied. The biomechanical analyses of aortic samples from BAPN-treated rats showed a significantly increased diameter (1.64 +/-0.02 mm), a significantly reduced maximum load (1.08+/-0.08 N), and a significantly reduced maximum stiffness (3.34+/-0.10 N) compared with controls (1.57+/-0.02 mm, 1.55+/-0.04 N and 4.49 +/-0.14 N, respectively). No changes in the concentrations of collagen and elastin were found. The content of pyridinoline, a mature collagen cross-link, was significantly decreased by 49% in the BAPN-treated group compared with controls. No changes in the concentration of desmosine + isodesmosine, the major cross-links of elastin. were found. The present study shows that cross-links are essential in providing mechanical stability of the aorta. Even a partial inhibition of the cross-linking processes results in a destabilisation of the aortic wall with increased diameter and reduced strength and stiffness. PMID- 9733225 TI - Soy lecithin reduces plasma lipoprotein cholesterol and early atherogenesis in hypercholesterolemic monkeys and hamsters: beyond linoleate. AB - The current study was designed to investigate the hypocholesterolemic and anti atherogenic properties of soy lecithin beyond its fatty acid content. In experiment 1, 18 cynomolgus monkeys were divided into three groups of six and fed diets which approximated either the average American diet (AAD), the American Heart Association (AHA) Step I diet, or a modified AHA (mAHA) Step I diet containing 3.4% soy lecithin for 8 weeks. Plasma samples were collected from food deprived monkeys and analyzed for total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), very low- and low-density lipoprotein cholesterol (non-HDL-C), and triglyceride (TG) concentrations. Group comparisons revealed that monkeys fed the mAHA Step 1 diet had significantly lower plasma TC (-46%) and non-HDL-C (-55%) levels compared to the AAD diet, whereas monkeys fed the AHA Step 1 diet had lesser reductions in plasma TC (-21%) and non-HDL-C ( 18%) levels. The monkeys fed the mAHA Step I diet had significantly lower plasma TC (-32%) and non-HDL-C (-45%) compared to the monkeys fed the AHA step diet. Also, only the mAHA Step I diet significantly reduced pre-treatment plasma TC and non-HDL-C levels by - 39 and -51% respectively with no significant effect on plasma HDL-C or TG levels. In experiment 2, 45 hamsters were divided into three groups of 15 and fed the following three modified non-purified diets for 8 weeks: a hypercholesterolemic diet (HCD) containing 10%, coconut oil and 0.05%, cholesterol, HCD plus 3.4%, soy lecithin (+SL), or the HCD with added levels of linoleate and choline equivalent to the +SL diet but no lecithin (-SL). Plasma lipids were determined as in experiment 1 and aortas were perfusion-fixed and Oil Red O stained for morphometric analyses of fatty streak area. Relative to the HCD group, the +SL-treated hamsters had significantly lower plasma TC (-58%), non-HDL C (-73%) and aortic fatty streak area (-90%). Relative to the -SL group, hamsters fed the +SL diet had significantly lower plasma TC (-33%), non-HDL-C (-50%) and significantly reduced aortic fatty streak area (-79%). In conclusion, the first experiment suggests that the cholesterol-lowering efficacy of the AHA Step I diet can be enhanced with the addition of soy lecithin without reducing plasma HDL-C levels. whereas the second experiment suggest that the hypocholesterolemic, and in particular, the anti-atherogenic properties of soy lecithin cannot be attributed solely to its linoleate content. PMID- 9733226 TI - Lipoprotein(a) and the significance of the association between platelet glycoprotein IIIa polymorphisms and the risk of premature myocardial infarction. AB - Platelet glycoprotein IIb/IIIa may be involved in the pathogenesis of myocardial infarction as the key element in platelet aggregation and as the binding site of lipoprotein(a) to platelets, inhibiting plasminogen binding and activation. Recently, a strong association between the P1A2 polymorphism of the glycoprotein IIIa gene and acute coronary thrombosis has been reported. although this has not been confirmed. In an associated study, we determined plasma lipoprotein levels, the apo E genotype and the P1A genotype in 250 males under 55 years with myocardial infarction and they were compared with 250 age- and sex-matched controls. Patients showed an over-representation of the epsilon3/4 genotype with respect to the control group. We found that there were no differences in the allelic frequency of P1A2 between case patients and age-matched controls (chi2 = 0.05, P = 0.92) and that subjects bearing the P1A2 allele showed higher plasma lipoprotein(a) concentration than p1A1/P1A1 individuals. Therefore, in this population there is no association between carriage of p1A2 allele and increased risk of myocardial infarction but the carriage of P1A2 is associated with higher plasma Lp(a) concentration. PMID- 9733227 TI - Association between angiotensin I-converting enzyme genotypes, extracranial artery stenosis, and stroke. AB - The insertion(I)/deletion(D) polymorphism of the angiotensin-converting-enzyme (ACE) gene has been associated with an increased risk of myocardial infarction, lacunar stroke, and with an increased intimal-medial thickness in several populations. The aim of this study was to evaluate whether the ACE I/D genotype is associated with stenosis of extracranial arteries and stroke in middle-aged and aged men and women. We studied 388 patients (247 male, 141 female) using Doppler and Duplex ultrasound of the extracranial arteries. Patients' history was obtained by standard questionnaire and by the hospital case records. Genomic DNA was analyzed by polymerase chain reaction (PCR) to identify the I/D polymorphism, with a second insertion specific PCR in samples classified as homozygous DD genotypes to prevent mistyping. The ACE genotype groups (DD 132, ID 164, II 92) were well matched for the basic characteristics. The DD genotype was more common in patients with extracranial artery stenosis > or = 50%, compared with patients without stenosis (59/147 versus 73/241, odds ratio 1.54, 95%-CI 1.01-2.37), but was not associated with a history of stroke (30/91 versus 102/297, odds ratio 0.94, 95%-CI 0.57-1.54). The association of the DD genotype with extracranial artery stenosis was also present in hypertensive subjects (n = 206, odds ratio 1.76, 95%-CI 0.99-3.17). In the whole group multiple logistic regression analysis revealed that the association of the DD genotype with extracranial artery stenosis was independent of age, gender, hypertension, hyperlipidemia, and diabetes. In conclusion, the ACE DD genotype is a weak risk factor for hemodynamically relevant stenosis of extracranial arteries, but not for stroke. PMID- 9733228 TI - Accelerated cholesteryl ester transfer in patients with essential hypertension and the effect of ramipril treatment. AB - Although the transfer of cholesteryl ester (CE) from high-density lipoprotein (HDL) to the apolipoprotein B-containing lipoproteins (very-low-density lipoproteins + low-density lipoproteins) has been shown to be abnormally increased in a number of conditions associated with increased cardiovascular risk, it has not been studied in patients with essential hypertension (EH). To determine whether subjects with EH have increased CE transport, CE transfer (CET) was estimated isotopically and lipoprotein lipid and phospholipid composition determined in a group of 14 untreated normolipidemic (triglycerides 116+/-46, cholesterol 185+/-30, HDL 38+/-10 mg/dl) otherwise healthy ethnically diverse EH subjects. CET was significantly increased in EH subjects compared to a similar group of normotensive controls (EH: k = 0.27+/- 0.09 vs. control k = 0.11+/-0.02: P < 0.01). Lipoprotein concentration and composition were comparable in the two groups and closely resembled that of an age- and sex-matched reference group. The abnormal increase in CET persisted (k = 0.25+/-0.12) after 3 months of treatment with the angiotensin converting enzyme (ACE) inhibitor ramipril without a change in either plasma or lipoprotein lipids. Thus, CET is increased in normolipidemic subjects with EH and is not affected by the ACE inhibitor ramipril. PMID- 9733229 TI - The relationship between apolipoprotein E, dementia, and vascular illness. AB - The purpose of this study was to concurrently assess the relationship of Apolipoprotein E (APOE) with both dementias and vascular illnesses in the very old. Nine hundred and fifty nine subjects (mean age 85 years) in a long-term care facility were genotyped and cognitively tested with the Mini Mental State Exam. All subjects were studied for the relationship of APOE with atherosclerotic heart disease, hypertension, or stroke without concomitant dementia. Four hundred fifty individuals met criteria for inclusion into one of the following groups: Alzheimer's disease (n = 318), vascular dementia (n = 49), or not demented controls (n = 83) and were investigated for the relationship between APOE and these diagnostic categories. APOE epsilon4 was not associated with atherosclerotic heart disease, hypertension, or stroke without concomitant dementia. The APOE epsilon3 allele was more common in men with atherosclerotic heart disease. In contrast, the APOE epsilon4 allele was more common in patients with Alzheimer's disease (22%) and vascular dementia (26%) than in not demented controls (7%). APOE epsilon4 is associated with dementias in the very old, whereas its relationship with either peripheral or central nervous system vascular disease without dementia is not as robust. PMID- 9733230 TI - Short term effects of omega-3 fatty acids on the radial artery of patients with coronary artery disease. AB - Long-term dietary omega-3 fatty acids improve coronary endothelial function in CAD patients, heart transplant recipients and diabetics. This study assessed whether short term omega-3 fatty acids affect radial artery function in CAD patients. A high resolution A-mode echotracking device (NIUS 02) was used to measure continuously, radial artery internal diameter at rest, during flow mediated vasodilation (FMD), during cold pressure test (CPT), and after sublingual glyceryl trinitrate (GTN). We studied 18 male CAD patients in a randomized, double blind, placebo controlled design. Between pre- and post intervention measurements 24 h apart, nine subjects received 18 g fish oil concentrate (6.4 g eicosapentaenoic acid and 3.9 g docosahexaenoic acid) and nine subjects 18 g placebo. In the placebo group correlation between both baseline diameters was 0.98; P < 0.001. Pre-intervention FMD was 7.5+/-5.6%, CPT mediated vasoconstriction was 3.8+/-2.5%, and GTN induced vasodilation was 15.7+/-9.8%. Vascular responses post-intervention showed no significant difference to pre intervention, there was no significant difference between both treatment groups. The radial artery does not seem to be an immediate target for vasodilatory actions of omega-3 fatty acids. PMID- 9733232 TI - Exon 4, and in particular codon 152 of the LDL receptor gene, is a hot spot for point mutations. PMID- 9733231 TI - A novel apolipoprotein E2 variant, E2Toranomon (Q187E), identified in a type III hyperlipoproteinemia patient with coronary atherosclerosis. PMID- 9733233 TI - Hearing improvement after resection of cerebellopontine angle meningioma: case study of the preoperative role of transient evoked otoacoustic emissions. AB - In a retrospective case study of a patient with a right-sided cerebellopontine angle mass lesion, transient evoked otoacoustic emissions were robustly present despite a severe to profound sensorineural hearing loss and abnormal auditory brainstem response. These results were interpreted as suggestive of a neural site of lesion, and the potential for planned, preserved, or improved hearing by a suboccipital surgical craniotomy was considered. A gross total resection was successful. Three years postoperatively, the patient has normal hearing sensitivity and word recognition ability. PMID- 9733235 TI - Recovery nystagmus revisited. AB - Recovery nystagmus (RN) describes a spontaneous nystagmus with a fast-phase beating toward the ipsilesional ear. The mechanisms underlying RN implicate central vestibular system compensation processes. The presence of RN is significant because it implies that function has returned from the affected peripheral vestibular system. A case is described where RN was recorded. The processes underlying RN are described. PMID- 9733234 TI - Monitoring and predicting ototoxic damage using distortion-product otoacoustic emissions: pediatric case study. AB - Young children undergoing cisplatin chemotherapy are known to be at risk for progressive sensorineural hearing loss. Early detection of such hearing loss is important for providing management options. However, in ill and/or young children, behavioral audiometry may not be sufficiently precise to detect the early stages of hearing loss. This case illustrates that distortion-product otoacoustic emissions (DPOAEs) may be an appropriate cross-check measure to supplement and confirm pediatric behavioral data. Perhaps more importantly, this study suggests that DPOAEs may have the potential to predict the earliest stages of progressive hearing loss before such changes are seen in audiometric thresholds. PMID- 9733236 TI - Bizarre "sawtooth" tympanogram in a patient with otitis media. AB - A patient evaluated for otitis media had a bizarre tympanogram with a "sawtooth" configuration. Acute otitis media with a pinpoint perforation was diagnosed. After resolution of the acute infection, a 2-mm diameter perforation remained and the presence of a patulous eustachian tube was documented. Tympanoplasty was eventually required, after which a normal tympanogram was recorded. Our interpretation of the tympanogram is discussed and the literature reviewed. PMID- 9733237 TI - Managing hearing loss in a patient with Alzheimer disease. AB - This case study reports the management of hearing loss in a patient with Alzheimer disease (AD) living at home with a spouse care giver. The report highlights the interaction between symptoms associated with AD and hearing loss and the lack of data regarding remediation of hearing loss in this population. Specifically, the case illustrates the modifications in evaluation and verification of the hearing aid fitting that may be advisable when working with patients with AD. The data for this patient illustrate some novel measurement techniques that may assist the professional in documenting the impact of treatment in this population. PMID- 9733238 TI - Large vestibular aqueduct syndrome: an overlooked etiology for progressive childhood hearing loss. AB - If a drastic change in hearing has occurred in a child following a minor head trauma, change in barometric pressure, or physical exertion, large vestibular aqueduct syndrome (LVAS) should be considered. Most audiologists are unaware of LVAS or do not suspect it, in part due to the presence of a conductive component. LVAS can be seen in conjunction with Mondini's dysplasia or may appear by itself and is easily identified by a computed tomography scan. We present five cases of LVAS and discuss the natural history, audiologic and imaging findings, and relevant literature. PMID- 9733239 TI - Antiviral therapy in a child with pediatric human immunodeficiency virus (HIV): case study of audiologic findings. AB - Over the past decade, much research has been conducted to determine the auditory consequences of human immunodeficiency virus (HIV) infection and acquired immune deficiency syndrome (AIDS). This research, primarily using adult patients, has focused on the involvement of the central auditory nervous system (CANS). Measures of auditory evoked potentials, particularly the auditory brainstem response (ABR), can document changes in the CANS as the disease progresses and during treatment with antiviral therapies such as zidovudine (AZT) and didanosine (ddI). This case study presents the audiologic findings for a child with HIV infection. Evaluations were performed over a 2-year period prior to the initiation of antiviral therapy and following treatment. Audiologic measures included behavioral audiometry, tympanometry, otoacoustic emissions, and ABR latency/intensity functions and rate studies. Findings indicated a gradual shortening of all ABR component latencies following the initiation of antiviral therapy. In addition, a high-frequency hearing loss was detected during the final evaluation subsequent to 19 months of treatment with AZT and ddI. PMID- 9733240 TI - Central auditory processing in a patient with bilateral temporal lobe tumors: case report. AB - The purpose of this article is to report the audiologic and central auditory processing abilities of a 34-year-old male with a right temporal lobe tumor and a history of bilateral tumors of the temporal lobes. The patient was evaluated presurgical re-exploration and again at 2.5 months and 4 months postoperatively. Test results demonstrated little change in peripheral hearing abilities; however, marked fluctuations were recorded on several tests administered postoperatively. Overall, this patient demonstrated a wide range of performance on tests of central auditory function, notably scores that decreased postoperatively and returned to better than baseline on the SCAN-A and repeated abnormal scores on the Pitch Pattern Sequence Test and the Symbol Digit Modality Test. Auditory Fusion Test-Revised results were initially normal, were markedly abnormal immediately postoperative, and returned to normal during the second postoperative visit. Our purpose in conducting this case study was to demonstrate, with central auditory processing test findings as well as magnetic resonance images, functional disorders of communication in a pre- and postoperative patient with a temporal lobe tumor. PMID- 9733241 TI - Auditory disorder in central nervous system miliary tuberculosis: case report. AB - We evaluated a 28-year-old female with a unilateral hearing loss of unusual pathogenesis, that of central nervous system miliary tuberculosis. Audiologic and otologic findings were consistent with left retrocochlear disorder, characterized by a profound hearing sensitivity loss, absent acoustic reflexes, normal otoacoustic emissions, and the presence of only wave I of the auditory brainstem response. Imaging studies revealed the presence of multiple punctate lesions, one of which was extra-axial and located at the left cerebellopontine angle. The pattern of audiometric test results, particularly the combination of normal otoacoustic emissions and profound hearing sensitivity loss, contributed importantly to the investigative sequence leading to the final diagnosis. PMID- 9733242 TI - Re: "Development of SCAN-A: test of auditory processing disorders in adolescents and adults" (J Am Acad Audiol 1996;6:286-292) PMID- 9733243 TI - Maurice H. Miller takes the editor to task. PMID- 9733244 TI - Parental perception of the adequacy of pain control in their child after discharge from the emergency department. AB - OBJECTIVE: To determine how well the pain of pediatric patients who are discharged from our emergency department (ED) is managed. DESIGN: Prospective, observational case series conducted from 9/21/96 to 3/16/97. SETTING: University tertiary care ED with an annual pediatric census of 11,000, consisting of a diverse racial and socioeconomic population. PATIENTS: Children ages < or = 15 years and discharged from the ED with one of the preselected acute, painful, conditions including fracture, corneal abrasion, ankle sprain with swelling, burn, otitis media with pain at discharge, or exudative pharyngitis. STUDY PROTOCOL: At time of discharge from the ED, data collectors not involved in the patients' care obtained consent from the patient's parent or guardian and completed data collection that included the final diagnosis and any recommended analgesic. Physicians were blinded to patient enrollment. Parents were phoned 48 hours after ED discharge and asked if they believed their child's pain was adequately controlled at home and if they had sought medical care elsewhere. Data were reported as percentages, and confidence intervals (CI) were calculated. RESULTS: From a convenience sample of 75 patients, five could not be contacted, leaving 70 for analysis. The mean age was 5.4+/-4.7 years. Sixty-seven parents (96%; 95% CI, 91-100%) believed their child's pain was well controlled, and 67 patients (96%; 95% CI, 91-100%) received an analgesic prescription. Five parents did not administer the prescribed analgesic because they believed their child's pain was controlled without it. No parent sought further medical care for pain medication for their child. CONCLUSIONS: Our pediatric pain management study showed high analgesic use and high parental satisfaction. Lectures and bedside education may be a way to improve pain management in pediatric patients. PMID- 9733245 TI - Pediatric ambulance utilization in a large American city: a systems analysis approach. AB - BACKGROUND: Research on utilization of ambulances by pediatric patients lacks an objective, reproducible tool for the evaluation of patterns of ambulance use by both the providers and the users of this resource. OBJECTIVES: 1) To develop an objective, diagnosis-based measure of appropriateness of ambulance utilization. 2) To use the measure to evaluate whether Municipal Ambulance Service dispatchers assign ambulances appropriately, and whether parents/caretakers request ambulances appropriately. STUDY DESIGN: 1) Development of the pediatric ambulance need evaluation (PANE) tool: The consensus of an expert panel was used to assign patients arriving by ambulance to three levels of prehospital transport need based upon their ultimate hospital discharge diagnoses, and were as follows: required advanced life support ambulance (ALS); required basic life support ambulance (BLS); required a less acute mode of transport (LAT). 2) Assessment of appropriateness of ambulance assignments by EMS call-receiving operators (CRO) and of ambulance requests by parents/caretakers: Comparison of actual type of ambulance assigned and of need for ambulance, using the PANE tool and hospital admission rates as gold standards. DATA COLLECTION: Level of prehospital transport provided (ALS vs BLS), ultimate ED diagnosis, and ED disposition (admission vs discharge) was collected for each patient from information abstracted from the prehospital and ED records. SETTING: Bellevue Hospital Center and Harlem Hospital Center, two level I trauma centers in New York City, both with Pediatric Emergency Departments staffed 24 hours a day by attending physicians and residents. PATIENT SELECTION: Consecutive sample of 2633 patients, birth to 18 years of age, who arrived to either hospital by ambulance as primary transports from the field over a one-year period. RESULTS: 1) Development of PANE tool: At Bellevue Hospital, 7% of ED visits arrived by ambulance; at Harlem Hospital, 5% arrived by ambulance. Using these ambulance arrivals, 215 diagnoses were identified for inclusion in the PANE tool. An expert panel categorized each diagnosis as requiring ALS, BLS, or LAT, with a high level of interobserver agreement (weighted kappa = 0.793). As a measure of external validity of the PANE, admission rates were highest in the ALS group, next highest in the BLS group, and lowest in the LAT group (chi2 for trend, P < 0.05). 2) Assessment of ambulance assignments and requests: According to the PANE tool, the sensitivity of dispatcher assignment of ALS ambulances was 72 %. Therefore, 28 % of patients who required an ALS ambulance received BLS care. 50% of patients assigned to an ALS ambulance did not require that level of care, and 1/3 of these were categorized by the PANE as not requiring an ambulance at all. CONCLUSIONS: The PANE tool compared favorably to admission rates as a measure of the severity of illness of patients arriving by ambulance. Applying the PANE tool, we conclude that the majority of requests for ambulances are appropriate, and that the majority of the time dispatchers were able to dispatch the appropriate level of care. However, there is room for significant improvement in utilization of ambulances, and tools like the PANE will be useful in achieving this goal. PMID- 9733246 TI - pH monitoring during diabetic ketoacidosis using the Paratrend 7 continuous blood gas monitoring system. PMID- 9733247 TI - Unexpected second foreign bodies in pediatric esophageal coin ingestions. AB - OBJECTIVE: To determine the frequency of unexpected second foreign bodies in children who present to the pediatric emergency department with esophageal coin impaction. DESIGN/METHODS: A retrospective chart review pediatric patients with esophageal coin impaction who underwent esophagoscopy/laryngoscopy for coin removal in a 16-year period at a tertiary referral center. Data analysis consists of descriptive statistics. RESULTS: Eighty three of 85 (95%) eligible charts were reviewed. Three children (3.6%) had unsuspected second foreign bodies: an adherent penny, a second penny low in the esophagus, and pieces of paper and lint. No significant esophageal injury occurred. CONCLUSIONS: Unexpected second foreign bodies in pediatric esophageal coin ingestions with adequate radiographic studies are rare and generally do not cause significant esophageal injury. PMID- 9733248 TI - Differentiation of systemic infection and congenital obstructive left heart disease in the very young infant. AB - BACKGROUND: The differentiation of severe systemic infection, such as sepsis or meningitis, from a congenital obstructive left heart abnormality presents a unique challenge to clinicians responsible for the care of such infants in the first few weeks of life. Clinical findings are very similar in the two populations. Failure to identify the need for specific intervention, such as prostaglandin administration, by the primary care or emergency physician may result in increased morbidity or death in these infants. METHODS: We undertook a retrospective review of critically ill infants 0 to 28 days of age presenting with either bacterial sepsis or meningitis or a congenital obstructive left heart syndrome (COLHS), in order to identify historical, physical, or laboratory findings which might differentiate the two groups at presentation. Discriminant analysis was performed using the presence or absence of COLHS as the dependent variable. A COLHS index was derived to determine its sensitivity and specificity for differentiating the two groups. RESULTS: The presence of cardiomegaly predicted COLHS with 85% sensitivity and 95% specificity. Cardiomegaly had a positive predictive value for COLHS of 0.95. Unfortunately, most of the other variables which, individually or in combination, were significantly different between the two groups demonstrated poor sensitivity for prediction of the presence of obstructive left heart disease. Eleven of the predictor variables were chosen for inclusion in the multivariate model, and a COLHS index was developed which correctly classified 62/63 cases (98% sensitivity, 100% specificity). CONCLUSIONS: We conclude that while it is very difficult to differentiate these two groups at presentation, early clinical suspicion of COLHS with attention to key clinical parameters identified in this study may expedite appropriate intervention and enhance outcome. The multivariate model derived may provide a template from which further research can elucidate a more clinically useful tool for the clinician. PMID- 9733250 TI - Lower extremity pain in a three year old: manifestations of Guillain-Barre syndrome. AB - Guillain-Barre syndrome (GBS) in children can present with early symptoms of limb and back pain that progress to acute generalized paralysis. Although the clinical course of this illness may be benign with spontaneous recovery, treatment is indicated for more severe cases. We report a three-year-old patient with GBS, whose symptoms were initially missed, and discuss the salient diagnostic and therapeutic issues for this disease. PMID- 9733249 TI - Childhood hypoglycemia in an urban emergency department: epidemiology and a diagnostic approach to the problem. AB - OBJECTIVE: To 1) determine the prevalence of hypoglycemia in childhood in a pediatric emergency department (ED), 2) determine epidemiology of idiopathic ketotic hypoglycemia (IKH), 3) determine diagnostic yield of the workup of hypoglycemia, and 4) review a diagnostic approach to hypoglycemia. SETTING: Urban pediatric ED of a tertiary level children's hospital. METHODS: Retrospective review of all medical records with a primary or secondary diagnosis of hypoglycemia (ICD-9 code 251.2) seen at the ED between 1/92 and 8/95. RESULTS: Thirty-one patients were identified. Mean blood glucose was 34.2 mg/dl. Prevalence of hypoglycemia among population seeking care in our ED was 6.54/100,000 visits. Eighteen patients were diagnosed with IKH for a prevalence of 3.9/100,000. IKH demographics were: mean age 27.7 months; 12 males, 6 females; 8 white, 9 black, and 1 not available. The weights of five patients were < 25th percentile. Fourteen of the 18 IKH patients had hormone studies done insulin [cost $40], growth hormone [$69], cortisol [$54]. All 14 had appropriately suppressed insulin levels (< 5microU/ml) and high cortisol levels > 22 microg/ml. Thirteen of the 14 had normal or high growth hormone (GH) levels (0.7-6 ng/ml). Four IKH patients had urine drug screens ($280); all were negative. Although no IKH patient was febrile, six had sepsis workups ($380); all were negative. Urine ketones were positive in 15 of the 18 tested (> 3+ in eight patients). Mean anion gap was 20 (range: 16-30). Eight of the 18 IKH patients were discharged from the ED after return to normal status. CONCLUSIONS: IKH is the most common cause of hypoglycemia in children beyond the infancy period. In its typical presentation (previously healthy one- to five-year-old, with normal growth and development, who presents with a first episode of symptomatic fasting hypoglycemia and appropriate degree of ketonuria, without hepatomegaly, and with resolution of symptoms on administration of glucose), an extensive and overzealous workup for endocrinopathy or inborn error of metabolism is not necessary. PMID- 9733251 TI - An unusual complication of endotracheal intubation: ingestion of a laryngoscope bulb. AB - Many well described complications can result from endotracheal intubation in neonates during resuscitation. Ingestion of a laryngoscope bulb is a rare event with potentially serious consequences. We are reporting this unusual complication in a neonate during delivery room resuscitation and point to the importance of checking the integrity of the equipment prior to resuscitative efforts. PMID- 9733252 TI - Pseudoparalysis of the lower extremity in an infant. AB - We describe a case of septic arthritis of the knee in an afebrile, well-appearing four-month-old female. She had been evaluated previously for lower extremity disuse attributed to antecedent trauma. Her physical examination was remarkable only for limitation of extension of the affected leg at the knee. Emergency physicians should understand the physical examination findings suggesting joint effusion and the need to consider osteomyelitis/septic arthritis in children less than one year of age with pseudoparalysis. PMID- 9733253 TI - Cocaine toxicity following dermal application of adrenaline-cocaine preparation. PMID- 9733254 TI - Early hemodynamic variations assessed by an echo-Doppler aortic blood flow device in a severely burned infant: correlation with the circulating cytokines. AB - We describe the case of a seriously burned infant who suffered from a deep burn covering approximately 30% of his total body surface area. Because invasive hemodynamic monitoring is usually not suggested in infants, hemodynamic profile can be misunderstood. We tested a new echo-Doppler device to determine hemodynamic variation using a small esophageal probe specifically designed for newborns and infants. The aortic flowmeter was connected with satellite devices to obtain the hemodynamic profile, including aortic blood flow (ABF), preejection period, left ventricular ejection time, mean arterial pressure, heart rate, calculated stroke volume, calculated total systemic vascular resistance (TSVR), and end-tidal CO2 pressure. The positioning of the probe was easily obtained at each time. The hemodynamic management initially exhibited a hypovolemic status followed by a hyperdynamic profile, as suggested by a gradually increased ABF, which seemed similar to the variations currently reported in adult burn patients. Concurrent with hemodynamic determinations, plasma samples were drawn to measure interleukin-6 (IL-6), interleukin-1beta(IL-1beta), and tumor necrosis factor alpha(TNF-alpha)levels. A consistent peak of IL-6 occurred simultaneously with the drop in TSVR. In contrast, no marked modifications were observed with IL 1beta and TNF-alpha. The same circulating cytokines moved alike in burned adults; IL-6 could partly explain the mechanisms of hemodynamic variation through the systemic inflammatory response syndrome. On the other hand, the echo-Doppler device could provide valuable noninvasive findings, allowing early improvement in resuscitation during the acute phase of critically burned infants and children. PMID- 9733255 TI - Conjunctival staining and corneal and conjunctival abrasions caused by 2% aqueous gentian violet solution. PMID- 9733256 TI - Tension pneumothorax secondary to grass head aspiration. AB - Aspiration of inflorescence or grass heads (seed head of grasses) often presents with atypical signs and symptoms because grass heads have a tendency to rapidly migrate to the periphery of the lung. If this is not recognized, it can lead to delay in diagnosis and serious complications. Removal with rigid bronchoscopy maybe difficult, and surgery is often needed. We report a case of a seven-month old child who had a delayed diagnosis of grass head aspiration and subsequently presented with a life threatening tension pneumothorax. This case highlights the importance of obtaining a detailed history in cases of foreign body aspiration and the need to include it in the differential diagnosis of unexplained respiratory symptoms, especially those of sudden onset in children. PMID- 9733257 TI - Mastoiditis: a case-based review. AB - Mastoiditis was a common complication of otitis media in the preantibiotic era. Because its incidence has decreased dramatically in recent years, its diagnosis can be delayed or missed, particularly in infants and young children. This delay can result in significant morbidity and increased costs, owing to longer inpatient treatment and surgical intervention. We offer a current, case-based review of the pathogenesis, presentation, and treatment of mastoiditis. PMID- 9733258 TI - Targeted management strategies for cardiovascular toxicity from tricyclic antidepressant overdose: the pivotal role for alkalinization and sodium loading. PMID- 9733259 TI - Five month old in a motor vehicle accident. PMID- 9733260 TI - Pediatric emergency medicine: legal briefs. PMID- 9733261 TI - The passive and aggressive evaluation of the cervical area. PMID- 9733262 TI - A case of acute choreoathetoid movements following a single ingestion of pemoline in a two-year-old boy. PMID- 9733263 TI - The psychosocial impact of bone marrow transplantation: a review of the literature. AB - Bone marrow transplant (BMT) is a procedure used for the treatment of a variety of cancers and malignant diseases. Recovery from this intensive process requires a long-term course, often accompanied by acute morbidity which includes various distressing physical symptoms. Recent literature has begun to explore the impact of this procedure on quality of life and psychosocial issues. While survivorship is often associated with a highly rated global quality of life, recovery from BMT is accompanied by several psychosocial difficulties which negatively impact patients. Fatigue is a common complaint, often hindering recipients for several years following their transplant. As well, reports of psychological distress, psychiatric symptoms, and/or mood disturbances such as anxiety or depression are not uncommon. Many patients also indicate interruption of sexual activity and increased sexual difficulty for several months following BMT. While some investigators have begun to examine hormone replacement therapy (HRT) as a treatment option for reducing sexual dysfunction, there is a general paucity of literature evaluating interventions for BMT survivors. This article reviews the literature examining various quality of life aspects including fatigue, psychosocial difficulties, and sexual functioning of patients during recovery from BMT. Limitations of past research are discussed and directions for future research suggested. PMID- 9733264 TI - Negative selection and protection of normal progenitor cells for autografting. AB - Autologous bone marrow transplantation (ABMT) after high-dose chemotherapy is recognized as a curative approach to treating hematologic malignancies and some invasive solid tumors. However, tumor cells present in the bone marrow at the time of harvesting are a potential cause for relapse. Ex vivo marrow purging with very high doses of cytotoxic agents has been introduced in an attempt to remove neoplastic cells contaminating the autograft. The procedure, however, has been limited by its high toxicity to normal bone marrow progenitor cells. In their purging procedures, investigators have used agents such as amifostine, originally developed to protect against the effects of radiation and chemotherapy. In this article, the appropriateness of protecting normal cells with amifostine during various purging procedures will be reviewed. PMID- 9733265 TI - Transplantation activities and treatment strategies in paediatric stem cell transplantation centres: a report from the EBMT Working Party on Paediatric Diseases. European Group for Blood and Marrow Transplantation. AB - To determine the current approach to stem cell transplantation (SCT) in centres which treat predominantly paediatric patients, a questionnaire was sent to 67 centres known by the EBMT registry to perform SCT mainly in children. Fifty-five centres from 19 countries responded. Forty centres (75%) started their transplantation activities between 1980 and 1992. Median number of transplants/centre was 95 (range 8-400). Median number of transplants/centre/year was 18 (range 5-85). On average, there was one physician responsible for seven SCT/year while one nurse was involved for a median of 1.7 SCT/year. Median four rooms/centre (range 1-17) were available for paediatric SCT. The most common isolation facilities were rooms with high efficiency particulate air filtration (HEPA). Eighty-two percent (45/55) of the centres performed allogeneic as well as autologous SCT, while 5% (three centres) offered exclusively allogeneic SCT and 13% (seven centres) used only autologous stem cell rescue. Stem cell source for allogeneic SCT was bone marrow in 87%, peripheral blood (PB) in 10% and umbilical cord blood in 3%. Donors were HLA matched related in 57%, mismatched related in 13%, and matched unrelated in 30% of allogeneic SCT. PB was the most commonly used stem cell source for autologous SCT (48%), followed by BM (41%) and the two together (11%). Data analysis revealed substantial differences in protective care, stem cell processing and transplantation procedures within the centres, irrespective of the country, centre size and transplant type. PMID- 9733266 TI - Survival, disease-free survival and adverse effects of conditioning for allogeneic bone marrow transplantation with busulfan/cyclophosphamide vs total body irradiation: a meta-analysis. AB - Randomized, prospective studies comparing BUCY to TBI conditioning regimens for allogeneic bone marrow transplantation have yielded conflicting results. We investigated the overall survival, the disease-free survival and the toxicities of BUCY vs TBI-based regimens by conducting a meta-analysis of all published, randomized, prospective trials comparing these regimens. Five studies were analyzed. We evaluated six endpoints: survival, disease-free survival, veno occlusive disease (VOD) of the liver, acute GVHD, chronic GVHD, and interstitial pneumonitis. We combined individual study results using a random effects model. Survival and disease-free survival were better with TBI-based regimens than with BUCY, but these differences were not statistically significant (survival odds ratio 1.4, 95% confidence interval 0.9-2.2, P = 0.09; disease-free survival odds ratio 1.2, 95% confidence interval 0.7-2.1, P = 0.44). A power analysis indicated that BUCY was unlikely to have a clinically relevant survival or disease-free survival advantage. The power analysis could not exclude the possibility of such an advantage for TBI-based regimens. A significantly greater incidence of VOD occurred with BUCY (odds ratio 2.5, 95% confidence interval 1.2-5.2, P = 0.02). For the other side-effects, there were no significant differences. We concluded that TBI-based regimens cause less VOD than BUCY and are at least as good for survival and disease-free survival. PMID- 9733267 TI - High-dose chemotherapy with autologous stem cell support in patients with responding stage IV breast cancer. AB - Ninety-four patients underwent high-dose chemotherapy with stem cell support for stage IV breast cancer. The high-dose chemotherapy consisted of the Stamp V regimen in all patients comprising cyclophosphamide, thiotepa and carboplatin (CTCb). Twenty-three patients received sequential high-dose therapies with the first consisting of high-dose melphalan and the second of Stamp V. Two patients died from chemotherapy-related complications resulting in a transplant-related mortality at 100 days of 2.2%. The progression-free survival at 3 years was 36% in patients with no evidence of disease at the first course of high-dose therapy compared with 17% in patients with remaining disease at time of the high-dose therapy (P = 0.03). There was no difference in overall survival between patients with no evidence of disease and other patients. The source of stem cells, single or double courses of high-dose therapy, positive selection of CD34+ cells, or number of involved sites had no influence on either progression-free survival or overall survival. Further studies of more intensive induction chemotherapy followed by high-dose therapy with stem cell support are indicated. PMID- 9733268 TI - Allogeneic transplantation of unmanipulated peripheral blood stem cells in patients with multiple myeloma. AB - In multiple myeloma (MM), allogeneic bone marrow transplantation may produce complete and durable responses, but is accompanied by significant transplant related mortality (TRM). To assess feasibility and possible advantages offered by the use of allogeneic, growth factor-primed PBSC instead of marrow, we analyzed the data of 10 patients with MM (IgG = 6, IgA = 1, BJ = 2, non-secreting = 1; stage II = 1, stage III = 8, plasma-cell leukemia = 1) who received an allogeneic transplant with PBSC. Their age ranged between 35 and 53 years (median 45). All were HLA-identical to their sibling donors. Prior to allograft, six patients received standard-dose chemotherapy (DAV or CY-Dexa) and four a sequential intensified scheme with autologous PBSC support. At the time of transplantation, three patients were in CR, three in PR, three had refractory disease, one progressive disease. Patients were conditioned with busulfan-melphalan (n = 9) or busulfan-cyclophosphamide (n = 1), and were allografted with unmanipulated PBSC obtained by apheresis after treatment with G-CSF alone (n = 6) or GM-CSF followed by G-CSF (n = 4). All patients engrafted, with 0.5 x 10(9)/l PMN and 50 x 10(9)/l platelets on (median) day 13. Four patients had > or =grade II acute GVHD (grade II in 3, grade III in 1). Following allograft, CR was achieved in 71% patients. Eight are currently alive, with six in CR at a median of 18.5 months (range 7-28) from the transplant. Two patients died, 1 and 4 months from the allograft, respectively, and one is alive with progression. A PCR analysis of IgH rearrangement showed that residual disease was no more molecularly detectable in four out of seven evaluated patients following allograft. The results suggest that PBSC may improve the therapeutic efficacy of allogeneic transplant in MM, not only by a reduction of TRM but also by an improvement of rate and quality of response. PMID- 9733269 TI - Post-chemotherapy and cytokine pretreated marrow stromal cell layers suppress hematopoiesis from normal donor CD34+ cells. AB - Marrow stromal layers were used to investigate the potential role of negative regulators produced by the marrow microenvironment as one potential cause of hematopoietic suppression after chemotherapy and cytokines. Stromal layers were established from marrow of normal or prechemotherapy donors and breast cancer patients after hematological recovery from one cycle of 5-fluorouracil, leucovorin, doxorubicin, and cyclophosphamide and GM-CSF or PIXY321 (GM-CSF/IL-3 fusion protein). Normal donor CD34+ cells were placed in contact with stromal layers, and the number of colony-forming units for granulocytes and macrophages (CFU-GM) was determined. There were 25-79% fewer CFU-GM in post-chemotherapy stromal layer cocultures than in no chemotherapy cocultures. With neutralizing antibody to TNF-alpha the number of CFU-GM in no chemotherapy and post chemotherapy stromal cocultures was, respectively, 96 +/- 7% (n = 5) and 142 +/- 8% (n = 5) of the number with no antibody treatment. PIXY321 and GM-CSF pretreated stromal layers also suppressed production of CFU-GM. Anti-TNF-alpha promoted an increase in CFU-GM numbers from GM-CSF, but not PIXY321, pretreated stromal cocultures. The results demonstrate that post-chemotherapy marrow stromal layers were deficient in supporting in vitro hematopoiesis and suggest that negative regulators induced by chemotherapy and cytokines may be one cause for this defect. PMID- 9733270 TI - The importance of CD34+/CD33- cells in platelet engraftment after intensive therapy for cancer patients given peripheral blood stem cell rescue. AB - The study was designed to determine whether the number of CD34+/CD33- cells given at autologous peripheral blood stem cell (PBSC) rescue after intensive therapy for cancer was a better predictor of platelet engraftment than the total number of CD34+ cells infused. Comparison between the total number of CD34+ cells/kg infused with the number of CD34+/CD33- cells/kg infused showed that, generally, 2 x 10(6) total CD34+ cells contained 1.38 x 10(6) CD34+/CD33- cells. There was poor correlation between the number of CD34+/CD33- and CD34+/CD33+ cells in the graft (r = 0.332). Engraftment times for platelets and neutrophils were evaluated in 68 patients. There was no significant difference between the times for platelets to reach >25 x 10(9)/l or neutrophils to reach >0.5 x 10(9)/l among patients who received > or <2 x 10(6) total CD34+ cells or > or <1.38 x 10(6) CD34+/CD33- cells although the latter was consistently the better predictor. Platelet recovery to >50 x 10(9)/l and >100 x 10(9)/l was delayed significantly in patients who received <1.38 x 10(6) CD34+/CD33-/kg infused (P < 0.02 and P < 0.05, respectively). The number of CD34+/CD33- cells/kg infused was a stronger predictor of platelet recovery than the total number of CD34+ cells infused (P < 0.05 for platelets >50 or >100 x 10(9)/l). Although platelet recovery was delayed significantly in patients who had <4 x 10(4) granulocyte-macrophage colony forming units (CFU-GM)/kg infused, the time delay between receipt of PBSCs and availability of the colony counts limits the use of this assay to patients who do not require stem cells to be given immediately. Our data suggest that the number of CD34+/CD33- cells given at PBSC rescue provide information about the quality of the graft necessary for long-term platelet engraftment. However, since the percentage of CD34+/CD33- cells shows considerable inter-patient variation, measurement of this cell population may be important in patients who experience poor stem cell mobilization or when a target dose of 2 x 10(6) total CD34+ cells/kg is not achieved. PMID- 9733271 TI - A comparative study on the efficacy of CD8-positive cells in enhancing allogeneic bone marrow engraftment: cell sorting vs microbead selection. AB - We have used a superparamagnetic microbead selection system to positively select a murine bone marrow CD8+ cell population. The functional ability of these cells to enhance allogeneic bone marrow engraftment was compared with that of fluorescence activated cell sorter purified CD8+ cells. The CD8+ cell population prepared by the microbead selection procedure was as effective as cell sorter purified CD8+ cells in enhancing T cell-depleted allogeneic bone marrow engraftment in lethally irradiated mice. Phenotypic characterization of these cells shows that most of these CD8+ cells express CD3 and the T cell antigen receptor complex. PMID- 9733272 TI - Urinary excretion and pharmacokinetics of acrolein and its parent drug cyclophosphamide in bone marrow transplant patients. AB - The urinary excretion and pharmacokinetics of acrolein (ACRO) and its parent drug cyclophosphamide (CP) were investigated in 16 randomly selected bone marrow transplant (BMT) recipients when CP was used for conditioning. Patients suffering from aplastic anemia (n = 3) received a 4-day course of CP at a dose of 50 mg/kg daily infused intravenously (i.v.) over 1 h. Patients with leukemia (n = 13) were given either a combination of busulphan followed by CP at a dose of 50 mg/kg infused i.v. over 1 h for 4 days, or CP at a dose of 60 mg/kg by i.v. infusion over 1 h daily for 2 days followed by total body irradiation. Serial plasma samples and urine were collected after the start of the first CP dose. CP was analyzed by capillary gas chromatography, whereas ACRO was measured in urine by liquid chromatography. The plasma concentration-time data for CP conformed to the two-compartment model and the mean and s.e.m. values of alpha, beta, Vss, total clearance, and renal clearance observed were 1.29 (0.31) h(-1), 0.17 (0.03) h( 1), 0.67 (0.13) l/kg, 0.14 (0.02) l/h x kg, and 0.0188 (0.0052) l/h x kg, respectively. The mean and s.e.m. values of fraction of CP excreted in the form of ACRO during this interval (fmu) and ratio of the 24-h urinary concentration of ACRO/creatinine (Cmu(n)) were 1.96 (0.35%) and 9.11 (2.19) microg of ACRO/mg of creatinine, respectively. Two patients developed hemorrhagic cystitis (HC). Each of these two patients excreted significantly (P < 0.01) more ACRO in the first and second 4-h urine collection periods. However, there was no significant difference in fmu or Cmu(n) of ACRO between either of these two patients and the rest. This suggests that the rate of appearance of ACRO in urine is more crucial for developing HC than the cumulative amount excreted. PMID- 9733274 TI - Comparison of a patient-controlled analgesia system with continuous infusion for administration of diamorphine for mucositis. AB - Mucositis remains an important problem following BMT and may delay discharge from hospital. Patient-controlled analgesia (PCA) systems have been reported to be of benefit in controlling BMT-associated mucositis. The present study comprised 65 patients (age range 16-68 years; 19 allografts, 29 peripheral blood stem cell autografts and 17 autologous bone marrow). Subjects were prospectively randomised to receive intravenous diamorphine for pain relief either by conventional continuous infusion (CI) or by PCA, using a Medex Walkman 440 delivery system. Each patient assessed his/her pain control and nausea daily by a visual analogue scale. Twenty-two patients did not require any diamorphine. Four patients required diamorphine for pain other than mucositis, and four patients failed PCA control. Of 35 assessable cases, no difference in pain control was noted between CI and PCA. However, PCA-controlled patients required significantly less diamorphine than CI controlled patients (mean, 131 +/- 23 mg for PCA vs 296 +/- 40 mg for CI; P = 0.001), and PCA required fewer days of diamorphine than CI (mean, 7.17 +/- 0.66 days for PCA, 9.00 +/- 0.65 days for CI; P = 0.03). Side effects were minimal and equivalent in the two arms. The findings suggest that PCA and CI offer equivalent control of the pain of BMT-associated mucositis, but PCA requires less total consumption and duration of diamorphine therapy. PMID- 9733273 TI - Safety of therapeutic anticoagulation in patients with multiple myeloma receiving autologous stem cell transplantation. AB - The use of autologous stem cell transplantation (ASCT) for the treatment of multiple myeloma is increasing. Anticoagulation may be required during ASCT for conditions such as Hickman line thrombosis. The safety of anticoagulation in patients receiving ASCT is unknown. We report a retrospective case-control study of the safety of therapeutic anticoagulation in patients with multiple myeloma receiving ASCT. We identified 10 patients who received therapeutic anticoagulation during ASCT. For each of the 10 cases identified, two matched controls were selected. As a primary endpoint, bleeding complications were assessed. Secondary endpoints included survival, length of hospital stay, transfusion requirements, grade 4 toxicity, and days to platelet engraftment. Bleeding complications were not significantly different between patients receiving anticoagulation and controls (P = 0.3). Three of 10 anticoagulated patients and two of 20 controls had a bleeding complication. Mortality during admission was similar (P = 1.0); one anticoagulated patient and one control died of sepsis. A trend towards increased median number of platelet transfusions in the heparinized patients was seen (27 vs 12 units, P = 0.055), reflecting the higher transfusion threshold chosen for the anticoagulated patients. The other secondary endpoints did not differ between patients and controls. In this case control study, bleeding was not significantly increased in the group receiving anticoagulation during ASCT. This group electively received more units of platelets than controls. Thus, therapeutic anticoagulation can be managed with minimal increased toxicity during ASCT. PMID- 9733275 TI - Detection of hepatitis G virus/GB virus C after allogeneic bone marrow transplantation. AB - Abnormal liver function before allogeneic BMT has been associated with VOD. Hepatitis G virus/GB virus C (HGV) is a recently discovered virus suggested to be a cause of non-A, non-B, non-C, non-D and non-E hepatitis. The aim of this retrospective study was to analyze the risk for liver complications and time to engraftment in patients infected with HGV. Fifty patients transplanted in 1995 were examined with RT-PCR for HGV on samples collected before, and between 3 and 6 months after BMT. Seven patients had HGV detected before BMT. No patient became infected during or early after the BMT. There were no differences in either pre- or post-transplant liver function abnormalities, VOD, or time to neutrophil engraftment in patients who did or did not have HGV detected before BMT. We conclude that the importance of HGV infection for the development of post transplant complications is limited. PMID- 9733276 TI - Oerskovia xanthineolytica: a new pathogen in bone marrow transplantation. AB - A 53-year-old woman with non-Hodgkin lymphoma underwent an autologous bone marrow transplant (BMT). Incomplete reconstitution necessitated the use of a long-term central venous catheter. One year after BMT she presented with fever. Echocardiography revealed vegetations on the tricuspid valve. Gram-positive rods grown from blood cultures and catheter tip were identified as Oerskovia xanthineolytica. We report the first case of native valve endocarditis caused by this organism. PMID- 9733277 TI - Lack of reactivity to CMV pp65 antigenemia testing in a patient with CMV disease following allogeneic bone marrow transplant. AB - Pre-emptive antiviral therapy based on the early detection of CMV infection is an important strategy for the prevention of CMV disease following allogeneic BMT. Accepted methods for early detection of CMV infection include viral culture of blood or bronchial lavage specimens or CMV pp65 antigenemia testing of peripheral blood specimens. We describe a patient with aplastic anemia with worsening liver transaminases after allogeneic bone marrow transplantation who had repeated negative tests for CMV pp65 antigenemia despite positive viral blood cultures. Re examination of peripheral blood samples with a different pp65 antibody pool revealed the presence of high levels of CMV in peripheral blood leukocytes, confirming a lack of reactivity to the original antibody pool. Following institution of antiviral therapy, a prompt reduction in the number of pp65 antigen-positive peripheral blood leukocytes paralleled a reduction in abnormal transaminases. The practical implications of these findings are discussed. PMID- 9733278 TI - Successful engraftment of unrelated cord blood stem cells for familial erythrophagocytic lymphohistiocytosis. Kinki Cord Blood Bank. AB - Familial erythrophagocytic lymphohistiocytosis (FEL) is an autosomal recessive disorder that can only be corrected by stem cell transplantation. One of our patients with FEL in second complete remission underwent successful cord blood stem cell transplantation (CBSCT); the donor was an HLA one-locus mismatched and unrelated individual. The conditioning regimen consisted of BU/CY/VP-16. The transfused cell dose was 6.8 x 10(7)/kg, which contained 1.36 x 10(5)/kg of CD34 cells and 3.4 x 10(4)/kg of CFU-GM. After CBSCT, there were no major infectious complications. Acute grade I GVHD was well controlled. Neutrophil counts reached >0.5 x 10(9)/l by day 20 and platelet counts reached >50 x 10(9)/l by day 40. Deficient natural killer activity returned to normal after the transplant. The patient recovered well more than 7 months after receiving CBSCT, without showing evidence of chronic GVHD. We recommend CBSCT for FEL patients who have no HLA matched siblings or unrelated donors. PMID- 9733279 TI - Rat pleural mesothelial cells adapted to serum-free medium as a model for the study of growth factor effects. AB - Mesothelial cells are the putative progenitors of mesotheliomas and cell lines have been used as tools to study the responses of these cells to various stimuli, including growth factors. The present study was undertaken to develop a rat mesothelial cell line capable of sustained growth under serum-free conditions with the object of avoiding the possible confounding effects of undefined serum components. Responses of mesothelial cells to epidermal growth factor were shown to differ under serum-free versus low-serum culture conditions. In contrast, a cell line, SFM1, adapted to growth in serum-free medium was characterized and found to exhibit responses to growth factors similar to the responses reported for human cell lines. This new line should prove to be a useful model for the study of these cells in vitro. PMID- 9733280 TI - Protection against cigarette smoke-induced damage to intact transformed rabbit corneal cells by N-acetyl-L-cysteine. AB - In order to assess cigarette smoke-induced oxidative damage to intact cells, an assay was developed to measure cell detachment and protection. Due to the complex nature of cigarette smoke, which contains molecules that can interfere with conventional spectrophotometric and fluorometric biochemical assays, transformed rabbit corneal cells were radiolabeled with tritiated thymidine and then subjected to direct stream smoke. As a result, cell damage in response to the smoke from only two cigarettes could be measured in a time-dependent manner. When cells were prelabeled with N-acetyl-L-cysteine (NAC), a substrate for glutathione synthesis, a significant reduction in damage was measured. Additionally, when buthionine sulfoximine (BSO), an inhibitor of glutathione synthesis, was incubated with cells, a reduction in the effectiveness of NAC was observed, although NAC still retained some activity. Furthermore, vitamin E conferred no protection to cells in this system nor was NAC active in a separate assay that appears to favor peroxyl radical generation. From these results we conclude that cigarette smoke damage can easily be determined at the cellular level with this technique and that NAC acted to prevent this damage in two ways: first, as glutathione precursor and, secondly, as an antioxidant capable of scavenging non peroxyl radicals. PMID- 9733281 TI - Sulfur mustard exposure enhances Fc receptor expression on human epidermal keratinocytes in cell culture: implications for toxicity and medical countermeasures. AB - Sulfur mustard (HD) is a chemical warfare blister agent. The biochemical basis of HD-induced vesication is unknown, and no antidote currently exists. Basal epidermal cells are a major site of HD toxicity in vivo, with inflammation and HD increased proteolytic activity implicated as factors that contribute to HD pathology. Fc receptors (FcR) bind to the Fc region of antibody to mediate many effector and regulatory functions that can influence inflammatory responses. FcR are found on all types of immune cells and are also expressed on the surface of human keratinocytes. Assay by fluorescent antibodies demonstrated significantly enhanced CD32 (FcRII) and CD16 (FcRIII) on human epidermal keratinocyte (HEK) cell cultures at 8 to 24 h after exposure to HD (50, 100 and 200 micromol/L). The enhanced CD32 was time- and concentration-dependent and agreed well with the time course of increased proteolysis and cutaneous pathology observed during HD vesication. HD-increased FcR on the surface of HEK might be a mechanism of vesication. PMID- 9733283 TI - In vitro study of cytotoxicity of quinolones on rabbit tenocytes. AB - Tendinitis and tendon rupture complicating fluoroquinolone therapy have been reported recently, especially affecting men over 60 years. These new quinolones are more potent antimicrobial agents than older nonfluorinated compounds like nalidixic acid. We compared the effects of one quinolone (nalidixic acid) and two fluoroquinolones (norfloxacin and pefloxacin) on cultured rabbit Achilles tendon cells. First, we examined their effects on cell viability, mitochondrial succinate dehydrogenase and global activity, mitochondrial activity using microtitration methods. Pefloxacin and norfloxacin were more cytotoxic than nalidixic acid according to IC50 values. These results confirm that mitochondria represent a biological target of fluoroquinolones. Moreover, the extracellular matrix was studied by molecular hybridization. After a 72 h treatment, the level of type I collagen transcripts was not modified with any of the three antimicrobial agents, whereas mRNA encoding decorin was decreased with 10(-4) mol/L pefloxacin only. The decrease of transcripts encoding decorin suggests that this matrix component is another target of pefloxacin and modification of decorin seems to be an early event (before mitochondrion alteration) which may contribute to the explanation of tendon rupture. PMID- 9733284 TI - Cooperative interaction of autocrine and paracrine mitogens for airway epithelial cells. AB - Mitogens of the EGF family may play an important role in regulating the proliferation of airway epithelial cells (AEC). We examined the production of autocrine mitogenic activity by mouse AEC cultured from explants of tracheal tissue. DNA synthesis by growth-arrested AEC was stimulated by conditioned media from cells maintained in serum-free culture without exogenous growth factors. The mitogenic activity was blocked by a specific inhibitor of the EGF receptor tyrosine kinase. Furthermore, conditioned media from AEC contained molecular species that could compete with radiolabeled EGF in a receptor binding assay. However, mitogenic activity was not blocked by neutralizing antibodies to EGF or to transforming growth factor-alpha, but was partly inhibited by co-incubation with heparin, suggesting that it might be due to a heparin-binding member of the EGF family. The activity was potentiated by co-incubation with IGF-1, analogous to the potentiation by IGF-1 of the mitogenic activity of EGF for AEC. Moreover, the autocrine mitogen produced by AEC exhibited cooperative interaction with the mitogenic activity in conditioned media from growth factor-deprived mouse lung fibroblasts, consistent with the hypothesis that interactions with mesenchymal cells could influence the proliferation of AEC in vivo. PMID- 9733282 TI - Single-strand breaks, cell cycle arrest and apoptosis in HL-60 and LLCPK1 cells exposed to 1,2-dibromo-3-chloropropane. AB - We investigated 1,2-dibromo-3-chloropropane (DBCP)-induced DNA damage, cell cycle alterations and cell death in two cell lines, the human leukemia HL-60 and the pig kidney LLCPK1, both of which are derived from potential target sites for DBCP induced toxicity. DBCP (30-300 micromol/L) caused a concentration-dependent increase in the levels of DNA single-strand breaks in both cell lines as well as in cultured human renal proximal tubular cells. After extended DBCP exposure in LLCPK1 cells (100 micromol/L, 30 h), the level of DNA breaks returned almost to control values. Incubation for 48 h showed a clear reduction of growth with DBCP concentrations as low as 10 micromol/L. Flow cytometric analysis showed that DBCP (1-10 micromol/L) exposure for 24 h caused an accumulation of LLCPK1 cells in the G2/M-phase. In HL-60 cells the accumulation in G2/M-phase was less marked, and at higher concentrations the cells accumulated in S-phase. Flow cytometric studies of HL-60 and LLCPK1 cells exposed to 100-500 micromol/L DBCP showed increased number of apoptotic cells/bodies with a lower DNA content than that of the G1 cells. Microscopic studies revealed that there were increased numbers of cells with nuclear condensation and fragmentation, indicating that apoptosis was the dominant mode of death in these cell lines, following exposure to DBCP. The characteristic ladder pattern of apoptotic cells was observed when DNA from DBCP treated HL-60 cells and LLCPK1 cells was electrophoresed in agarose. The finding that DBCP can cause an accumulation of cells in G2/M-phase and induce apoptosis in vitro may be of importance for the development of DBCP-induced toxicity in vivo. PMID- 9733285 TI - Modulation of adrenal cell functions by cadmium salts. 5. Cadmium acetate and sulfate effects on basal and ACTH-stimulated steroidogenesis. AB - In vitro and in vivo cadmium toxicity studies focus almost exclusively on CdCl2 effects. Only a few studies have used adrenocortical cells and tissue to determine cadmium salt effects during stress of adrenocorticotropin stimulation. Because several biologically relevant water-soluble cadmium salts exist, this study extended work with CdCl2 to evaluate the acute adrenocortical cell steroid secretory responses to non-lethal cadmium acetate (CdAc2) and CdSO4 concentrations. Control or ACTH-stimulated cultured Y-1 mouse adrenal tumor cells (ATCC) which secrete 20alpha-dihydroprogesterone (20-DHP) were incubated for 0.5 h in serum-free medium (FMEM) with or without 0.5, 1.0, 5.0, 10.0, 50.0, 100.0, 500.0 and 1000.0 microg CdAc2 or CdSO4/ml FMEM (1.9, 3.8, 19.0, 38.0, 190.0, 380.0 and 1900.0 micromol/L, respectively). For each salt, cell viability was measured at the end of the incubation using live cell trypan blue exclusion. In addition, cumulative CdAc2 effects during 4 h incubations and effect reversibility were determined for control and stimulated cells. After each experimental incubation, the 20-DHP secreted into the medium was determined by radioimmunoassay. Over 80% of all control or ACTH-stimulated cells were viable after incubation in the presence or absence of various CdAc2 or CdSO4 concentrations. Cadmium acetate and sulfate inhibited basal and ACTH-stimulated steroid secretion in a dose-dependent manner. For basal steroid secretion the CdAc2 concentration that first significantly inhibited was 0.5 microg/ml medium (1.9 micromol/L); stimulated secretion was significantly inhibited beginning at 5.0 microg/ml (19.0 micromol/L) and the concentration reducing stimulated 20-DHP secretion by 50% (IC50) was 5.6 microg/ml (21.3 micromol/L). Similarly, the first CdSO4 concentration to significantly inhibit basal and ACTH-stimulated steroid secretion was 10.0 microg/ml medium (39.0 micromol/L); the IC50 was 7.8 microg/ml (29.8 micromol/L). Except that basally secreting Cd2+-treated cells almost doubled 20-DHP secretion after Cd2+ removal and subsequent incubation with ACTH, all basal and ACTH-stimulated steroid secretion was irreversibly inhibited by every CdAc2 concentration. All CdAc2 concentrations initiated and maintained cumulative inhibitory effects on basal and ACTH-stimulated steroid secretion over a 4 h period. Reversibility and cumulative CdSO4 treatment studies were not conducted. Based on the results from the present studies, both CdAc2 and CdSO4 appeared to incrementally inhibit control and ACTH-stimulated steroidogenesis without affecting cell viability and to be more potent inhibitors of adrenocortical cell steroid secretion than CdCl2. Finally, CdAc2 effects on control and stimulated cells were cumulative and irreversible. PMID- 9733286 TI - PANDAS: the search for environmental triggers of pediatric neuropsychiatric disorders. Lessons from rheumatic fever. AB - Pediatric autoimmune neuropsychiatric disorders associated with streptococcal infection (PANDAS) is a relatively new diagnostic construct applied to children or adolescents who develop, and have repeated exacerbations of, tic disorders and/or obsessive-compulsive disorder following group A beta-hemolytic streptococcal infections. The proposed pathophysiology is that the group A beta hemolytic streptococcal bacteria trigger antibodies that cross-react with the basal ganglia of genetically susceptible hosts leading to obsessive-compulsive disorder and/or tics. This is similar to the etiologic mechanisms proposed for Sydenham's chorea, in which group A beta-hemolytic streptococcal antibodies cross react with the basal ganglia and result in abnormal behavior and involuntary movements. When first proposed, there was much controversy about the idea that streptococcal infections were etiologically related to rheumatic fever. In a like manner, discussion has arisen about the concept of infection-triggered obsessive compulsive disorder and tic disorders. We review the historical background to these controversies, give an update on the findings provided by research on PANDAS, and address areas of future study. PMID- 9733287 TI - Frequency of neurologic disorders in the neonatal intensive care unit. AB - Neonatal intensive care unit survival rates have improved significantly over the past decade. This improvement primarily reflects declining mortality rates among preterm infants. Neurologic morbidity increases with prematurity and is the major predictor of long-term disability. Accordingly, concern has been expressed that the burden of neurologic dysfunction among contemporary neonatal intensive care unit survivors may be increasing. To define the trends of neurologic disorders in the contemporary neonatal intensive care unit, all 4164 admissions between 1986 and 1995 to a tertiary neonatal intensive care unit were examined. Neonatal intensive care unit admissions (413 +/- 49 per year), proportion of births at less than 37 weeks (70 +/- 3% per year), and referral patterns were stable between 1986 and 1995. Over the study period, 773 (18%) of 4164 neonatal intensive care unit infants had a total of 1062 neurologic disorders. The neonatal intensive care unit mortality rate declined from 12% in 1986 to 4.2% in 1995 (P < .01). Neurologic disorders declined, from 27% of infants born in 1986 to 12% in 1995 (P < .001): 356 had seizures (14% in 1986 to 4% in 1995; P < .001), 235 had hypoxic-ischemic encephalopathy (8% in 1986 to 4% in 1995, P < .01), and 167 had intraventricular hemorrhage (7% in 1986 to 1.4% in 1995, P < .005). Frequency of congenital or chromosomal aberration affecting the nervous system was relatively constant (4.5% per year). Despite a three-fold improvement in neonatal intensive care unit survival between 1986 and 1995, the frequency of perinatally acquired neurologic disorders declined by more than 50%. PMID- 9733288 TI - Neurotrophic factors in cerebrospinal fluid and serum of patients with Rett syndrome. AB - Rett syndrome is now considered to be a neurodevelopmental disease. Its cause is unknown, but it has been suggested that neuronal growth factors and neurotransmitters play important roles. We measured levels of brain-derived neurotrophic factor and glial cell line-derived neurotrophic factor in cerebrospinal fluid, and nerve growth factor and brain-derived neurotrophic factor in serum in child and adolescent patients with Rett syndrome. Levels of brain-derived neurotrophic factor and glial cell line-derived neurotrophic factor in cerebrospinal fluid were below the limit of sensitivity of the methods used. Serum levels of nerve growth factor and brain-derived neurotrophic factor did not differ from control values. In Rett syndrome, the normal serum levels of nerve growth factor together and previously reported low levels of the factor in cerebrospinal fluid indicate that the latter may reflect low levels of nerve growth factor in the central nervous system. PMID- 9733289 TI - Evaluation of cerebellar size in attention-deficit hyperactivity disorder. AB - Evidence from animal and human research suggests that the cerebellum may play a role in cognition. This includes domains of executive function that are normally attributed to the prefrontal cortex and are typically deficient in individuals with attention-deficit hyperactivity disorder (ADHD). To investigate cerebellar structure in ADHD, magnetic resonance imaging morphometry was used to measure the area of the cerebellar vermis in 12 males with ADHD and 23 male controls matched for age and Wechsler Full-Scale IQ. Analyses were conducted to evaluate group differences, as well as differences between matched pairs of subjects with ADHD and those without ADHD. All measurements were corrected for overall brain size. Both analyses revealed that the size of the posterior vermis was significantly decreased in males with ADHD (P < .05 in both analyses), and that within the posterior vermis, the inferior posterior lobe (lobules VIII-X) was involved in this reduction (P < .05 for group analysis, P < .005 for matched pair analysis), while the superior posterior lobe (lobules VI/VII) was not involved in the reduction. The finding of abnormal inferior posterior vermal size suggests that dysfunction within this region of the cerebellum may underlie clinical deficits seen in individuals with ADHD. PMID- 9733290 TI - Respiratory sinus arrhythmia in children with severe cyanotic and pallid breath holding spells. AB - In this study we investigated centrally mediated parasympathetic regulation of modulated cardiac vagal tone among children with severe cyanotic and pallid breath-holding spells by examining respiratory sinus arrhythmia. Respiratory sinus arrhythmia was evaluated in 41 children; 17 subjects with cyanotic breath holding spells (6 boys, 11 girls; mean age 37.1 months), 7 subjects with pallid breath-holding spells (2 boys, 5 girls; mean age 33.0 months), and 17 controls (8 boys, 9 girls; mean age 41.2 months). Subjects had recurrent (more than 3) severe breath-holding spells. Each subject's electrocardiogram was recorded in a quiet room and digitized by an 80386 personal computer during five 1-minute periods. R R intervals within each 1-minute period were converted to heart rate in 120 successive 0.5-second intervals. The resultant heart rate time series was converted to its underlying frequency composition by a fast Fourier transform and averaged across minutes. Respiratory sinus arrhythmia was defined as the variability in the time series over a frequency range (0.096 Hz to 0.48 Hz) corresponding to a range of respiratory rates from 6 to 30 breaths per minute. ANCOVA adjusting for age and sex was conducted with the subject group as the independent measure. There were no significant differences between subjects with cyanotic breath-holding spells and controls. Pallid breath-holding spell subjects had a marked difference in respiratory sinus arrhythmia from either controls or subjects with cyanotic breath-holding spells, demonstrating less variability in respiratory sinus arrhythmia (P < .042) This study supports the hypothesis that there exists autonomic dysregulation in pallid breath-holding spells, caused by a primary central parasympathetic disturbance distinct from the dysregulation found in cyanotic breath-holding spells. PMID- 9733291 TI - Analgesic rebound headache in children and adolescents. AB - For more than a decade, the frequent use of analgesics has been recognized to lead to daily headaches in adults. To date, no studies on the occurrence of analgesic rebound headache have been done on the pediatric population. We retrospectively reviewed all charts of patients with the diagnosis of headache seen in our pediatric headache clinic between January 1996 and May 1997. Among the 98 patients seen, 46 (47%) suffered from daily or near daily headaches; 30 of them were consuming daily analgesics. Twenty-four patients (mean age 12.1 years, and mean follow-up 6.2 months) successfully discontinued their analgesics. Twenty two patients were also placed on amitriptyline. A significant reduction in the frequency (80%), severity (47%), and number of school days missed (74%) were seen. In conclusion, this data is comparable to previous observations reported in adults, and suggests that the daily use of analgesics might result in daily or near daily headaches in the pediatric population. Discontinuing daily analgesics, with the concomitant use of amitriptyline, is an effective treatment for analgesic rebound headache in this population. PMID- 9733292 TI - Rett syndrome: significant clinical overlap with Angelman syndrome but not with methylation status. AB - Rett syndrome and Angelman syndrome are neurodevelopmental disorders characterized by severe intellectual disability, microcephaly, speech disturbance, movement disorders with gait and/or truncal ataxia, and occasionally a similar facial appearance. Both conditions can be difficult to diagnose in girls early in their clinical course and can be difficult to distinguish from each other. Genomic imprinting is a known association in Angelman syndrome and previously has been suggested in Rett syndrome. Our aim was to evaluate the methylation status in a cohort of classical patients with Rett syndrome, using a methylation system for chromosome 15q11-13. Methylation analysis of chromosome 15 has not been previously reported in Rett syndrome. Furthermore, we document the clinical features of 31 girls with classical Rett syndrome and confirm the phenotypic similarities between Rett syndrome and Angelman syndrome. The methylation studies in these girls with Rett syndrome were normal. This excludes an imprinting error of the Angelman syndrome critical region on chromosome 15 (15q11-13) as an association with Rett syndrome, and indicates that methylation studies may be useful in distinguishing Rett syndrome from Angelman syndrome in young patients with an overlapping clinical phenotype. A normal methylation pattern, however, does not exclude the diagnosis of Angelman syndrome and clear distinction between the two syndromes will evolve over time. PMID- 9733293 TI - Cognitive neuropsychological and regional cerebral blood flow study of a developmentally dyslexic Japanese child. PMID- 9733294 TI - Pachygyria associated with childhood-onset epileptic spasms. PMID- 9733295 TI - Judicious resection and/or radiosurgery for parasagittal meningiomas: outcomes from a multicenter review. Gamma Knife Meningioma Study Group. AB - BACKGROUND: Parasagittal meningiomas, especially when associated with the middle or posterior third of the superior sagittal sinus, pose difficult management challenges. Initial surgical excision is associated with high morbidity and frequent tumor recurrence after subtotal resection. Neurological deficits are cumulative when multiple resections are required. No consistent management approach exists for patients with such tumors. In addition to observation, management options include resection, stereotactic radiosurgery, or fractionated radiation therapy used alone or in combination. METHODS: Sixteen centers where resection, gamma knife radiosurgery, and/or radiation therapy were available provided management data on 203 patients with histologically benign meningiomas from the time of initial diagnosis through follow-up after radiosurgery. The timing of resections, parameters of radiosurgery, rates of tumor control, morbidity, and functional patient outcomes were studied. The median follow-up duration in this study was 3.5 years (maximum, 33 yr after presentation and 6 yr after radiosurgery). RESULTS: The tumors were located in the anterior superior sagittal sinus in 52 patients, at the middle of the sinus in 91, and at the posterior portion of the sinus in 60. The mean tumor volume at the time of radiosurgery was 10 cc. In patients who underwent radiosurgery as the primary therapy (n = 66), the 5-year actuarial tumor control rate was 93 +/- 4%. No clinical failure (need for additional therapy or worsened neurological function) occurred in patients who had smaller tumors (<7.5 cc) and who had never undergone resection (n = 41). The 5-year control rate for patients with previous surgery was only 60 +/- 10%; the control rate for the radiosurgery-treated volume was 85%. Most failures resulted from remote tumor growth. Multivariate analyses identified significantly decreased tumor control with increasing tumor volume (P = 0.002) and previous neurological deficits (P = 0.002). The rate of transient, symptomatic edema after radiosurgery was 16%, was more common with larger tumors, and occurred within 2 years. Of 33 patients who were employed at the time of radiosurgery for whom a minimum of 1 year of follow-up data were available, 30 remained employed (91%). A decrease in functional status after radiosurgery was noted in only 3 of 33 (9%) employed and 7 of 77 (9%) unemployed patients. CONCLUSION: In patients with smaller tumors (<3 cm in diameter) and patent sagittal sinuses, we advocate radiosurgery alone as the first surgical procedure. Patients with larger tumors and those with progressive neurological deficits resulting from brain compression should first undergo resection. Planned second stage radiosurgery should be performed soon afterward for any residual tumor nodule or neoplastic dural remnant. Multimodality management may enhance long term tumor control, reduce the need for multiple resections, and maintain the functional status of the patient. PMID- 9733296 TI - Occipital lobe vascular malformations: prevalence of visual field deficits and prognosis after therapeutic intervention. AB - BACKGROUND: The prevalence of visual field (VF) deficits in association with vascular malformations of the occipital lobe is not known, and the prognosis of the VF after therapeutic intervention has not been systematically documented. METHODS: We reviewed the clinical records of 23 consecutive patients who were managed at a single institution during a 3-year period with intracranial vascular malformations extending within the anatomic borders of the occipital lobe. Lesion location and treatment rendered were correlated with formal VF testing performed before and after therapeutic intervention. RESULTS: Twenty-one of the 23 patients underwent treatment of their lesions. Treatment included resection after preoperative embolization (12 patients), resection alone (2 patients with arteriovenous malformations and 3 patients with cavernous malformations), and stereotactic radiosurgery (4 patients; preceded by embolization in 3 of the 4). One patient was followed expectantly, and another died as a result of hemorrhage soon after undergoing endovascular embolization. The VFs were assessed before and after therapeutic intervention (follow-up assessment, 2-29 mo). New VF deficits or worsening of preexisting VF deficits were documented in 5 of the 21 treated patients (24%), but only 2 of these patients (9.5%) had persistent deficits at the time of their follow-up examinations. Among the 10 patients with pretreatment VF deficits, 5 improved and the other 5 were unchanged after treatment. CONCLUSION: Patients with occipital lobe vascular malformations frequently present with associated VF deficits. Surgical resection or stereotactic radiosurgery (with or without previous embolization) of these lesions can be performed with little risk of causing new VF deficits or worsening of preexisting ones. Many VF deficits can be expected to improve or resolve after therapy. PMID- 9733297 TI - Radical excision of intramedullary cavernous angiomas. AB - OBJECTIVE: This is a retrospective study of patients with surgically treated, intramedullary cavernous malformations. We conducted the study to elucidate the outcomes of the patients, as well as potential pitfalls in their care. METHODS: A series of 12 patients underwent radical excision of intramedullary cavernous malformations between 1986 and 1996. All lesions were diagnosed by magnetic resonance imaging. Although seven patients experienced recurrent episodes of pain and sensorimotor disturbances, the histories of the other five patients were relevant for slowly progressing deficits (mostly sensory). RESULTS: All cavernomas were completely resected. No deaths were recorded. In follow-up examinations (5-102 mo after discharge), there was no evidence of recurrence, either clinically or in control magnetic resonance imaging scans. In follow-up examinations, two patients demonstrated sensory deficits that were slightly more pronounced than the preoperative deficits. The postoperative neurological status of 3 of 12 patients was unchanged, compared with the preoperative status. The status of the remaining seven patients had improved. For four patients there was effective functional improvement, and for three others there was complete postoperative relief of pain. Deficits of the long tracts were less prone to recover. CONCLUSION: The clinical course of cavernous malformations may be difficult to distinguish from that of spinal dural arteriovenous malformations or focal demyelinating disease. In the latter case, even magnetic resonance imaging results could be deceptive. Radical resection of these malformations is feasible, with relatively low surgical morbidity, provided that the preoperative deficits of the patients are limited. Given the generally progressive course of the illness and the few acute catastrophic myelopathies, complete excision is advocated whenever malformations are symptomatic. PMID- 9733298 TI - Reassessment of the role of radiation therapy in the treatment of endocrine inactive pituitary macroadenomas. AB - OBJECTIVE: This prospective clinical trial was undertaken to assess the rate of tumor recurrence in patients with endocrine-inactive pituitary macroadenomas who underwent gross total surgical resection of their tumors and did not receive adjuvant radiotherapy. METHODS: Between December 1987 and July 1994, 45 patients with endocrine-inactive pituitary macroadenomas underwent transsphenoidal surgery. In 38 (84%) of these patients, gross total surgical resection was achieved and was confirmed by postoperative magnetic resonance imaging (n = 37) or computed tomography (n = 1). After receiving counseling from the neurosurgeon concerning the risks and benefits of radiation therapy, 32 of the 38 patients elected not to receive adjuvant radiotherapy. Patients were followed through March 1998 with radiographic imaging obtained every 6 months for the first 2 years, annually for postoperative Years 3 and 4, and then every 2 to 3 years thereafter. The study end point was defined as radiographic tumor recurrence or patient death. RESULTS: The mean follow-up duration for the study group was 5.5 years. During that time, 2 of 32 (6%) patients developed recurrence, at 18 and 24 months, respectively, after initial surgery. Both were successfully treated using radiation therapy, with one requiring additional surgery. Three additional patients died as a result of unrelated causes 9, 12, and 49 months, respectively, after initial surgery. Immunocytochemical analysis revealed 66% of the tumors to be weak gonadotroph cell adenomas, 22% to be null cell adenomas, 9% to be silent prolactinomas, and 3% to be silent corticotroph cell adenomas. CONCLUSION: This study demonstrates a 6% 5-year recurrence rate in patients with endocrine inactive pituitary macroadenomas treated using gross total surgical resection alone. Reserving radiation therapy for the infrequent patient with recurrence and sparing the majority of patients the associated risks inherent in its use seems reasonable. PMID- 9733299 TI - Postoperative sella: evaluation with fast spin echo T2-weighted high-resolution imaging. AB - OBJECTIVE: The purpose of this study was to investigate the magnetic resonance features of the postoperative sella with fast spin echo (FSE) T2-weighted high resolution imaging and to evaluate the benefits of the sequence using a follow-up magnetic resonance imaging protocol after transsphenoidal surgery. METHODS: Coronal spin echo (SE) T1-weighted and FSE T2-weighted images were prospectively obtained in 24 patients after surgery for pituitary adenomas. We observed the signals and the contour of normal structures, fluid collection, implanted materials, and mass lesions, including granulation tissue and adenoma. RESULTS: The pituitary gland was delineated in 51 of 59 FSE T2-weighted images, 90.2% of which presented clear boundaries. Whereas the gland was detected in 49 of 58 SE T1-weighted images, only 20.4% showed the boundaries. A mass lesion was identified in each of 12 patients with good resolution on FSE T2-weighted images. SE T1-weighted images detected mass lesions in 7 of 12 patients without distinctive boundaries. Contrast enhancement had little advantage in clarifying the boundaries between normal and abnormal structures. For the detection of mass lesions in the sella, the kappa values for interobserver agreement were 0.8 for FSE T2-weighted images and 0.25 for SE T1-weighted images. CONCLUSION: FSE T2 weighted imaging is a reliable method with which to assess the sella with sufficient resolution after transsphenoidal surgery. The combination of unenhanced SE T1-weighted and FSE T2-weighted images may reduce the use of contrast material after pituitary surgery. PMID- 9733300 TI - Maternal shunt dependency: implications for obstetric care, neurosurgical management, and pregnancy outcomes and a review of selected literature. AB - OBJECTIVE: Because more women with cerebrospinal fluid shunts are surviving to child-bearing age, neurosurgeons, obstetricians, and other health care professionals require information about the care of these patients, especially during pregnancy and delivery. The purpose of this study was to gather comprehensive data from women with shunts regarding their clinical histories during and immediately after pregnancy. The following questions were addressed. 1) How does maternal shunt dependency influence the course of pregnancies and pregnancy outcomes? 2) What neurosurgical complications characterize this population of patients? 3) What complications of shunt dependency influence obstetric management, including prenatal testing and delivery? METHODS: A total of 37 respondents (age, 18-41 yr), accounting for 77 pregnancies, completed a questionnaire providing information on maternal background and medical history, shunt performance during pregnancy, management of delivery, pregnancy outcomes, and unusual complications. RESULTS: Fifty-six pregnancies resulted in live births; of these, 47 occurred in women with ventriculoperitoneal shunts. Three women underwent therapeutic abortions, 1 experienced preterm delivery, and 8 experienced 17 miscarriages. Four women experienced seizures during pregnancy, five reported third-trimester headaches, and eight described abdominal pains during the first and third trimesters. Four babies were diagnosed as having congenital defects. Shunt malfunctions and revisions occurred 10 times in 7 women, either during pregnancy or within 6 months after delivery. No acute malfunctions occurred during delivery. Forty-seven cases, representing 84% of all pregnancies, exhibited no shunt malfunctions or revisions. CONCLUSION: This study extends previous observations to a larger population of shunt-dependent mothers. The results suggest that maternal shunt dependency entails a relatively high incidence of complications but that proper care of these patients can lead to normal pregnancies and deliveries. PMID- 9733301 TI - Relief of trigeminal neuralgia after percutaneous retrogasserian glycerol rhizolysis is dependent on normalization of abnormal temporal summation of pain, without general impairment of sensory perception. AB - OBJECTIVE: This study was undertaken to examine the pathophysiological mechanisms of trigeminal neuralgia and the mechanisms underlying pain relief after percutaneous retrogasserian glycerol rhizolysis (PRGR). METHODS: Quantitative examination of sensory and pain perception was performed in the trigger area and the contralateral nonpainful facial skin area for 39 patients with trigeminal neuralgia who had been previously treated with PRGR and for 14 non-surgically treated patients. In a prospective study, 9 of the 14 patients were examined before and 4 to 6 weeks after PRGR. RESULTS: In the trigger area of patients who had been previously treated with PRGR for trigeminal neuralgia, we demonstrated increased temperature and tactile thresholds in pain-free patients and in patients with paroxysmal or continuous pain. Abnormal temporal summation of pain (characterized by progressive increases in pain intensity, with radiation of pain and aftersensation) was present in patients with paroxysmal or continuous pain but not in pain-free patients. In the trigger area of non-surgically treated patients with trigeminal neuralgia, we demonstrated significantly increased temperature and tactile thresholds and the presence of abnormal temporal summation of pain. The prospective study showed that pain relief after PRGR was associated with normalization of abnormal temporal summation of pain, without increased sensory loss. CONCLUSION: Partial deafferentation, with impairment of thin (C/Adelta) and thick (Abeta) fiber-mediated sensations and abnormal temporal summation of pain, is present in the trigger area of patients with trigeminal neuralgia. Relief of pain after PRGR depends on the normalization of abnormal temporal summation of pain, which is independent of general impairment of sensory perception. Assessment of the temporal summation of pain may serve as an important tool to record central neuronal hyperexcitability, which may play a key role in the pathophysiological changes in trigeminal neuralgia. PMID- 9733302 TI - Vestibular schwannoma management in the next century: a radiosurgical perspective. AB - PURPOSE: To discuss how the evolution of vestibular schwannoma radiosurgery, changes in health care delivery, and patient accessibility to medical information will affect the management of vestibular schwannomas in the future. CONCEPT: In comparison with microsurgical resection of vestibular schwannomas, radiosurgery has a lower morbidity rate, a similar risk of requiring further surgery, and higher patient satisfaction. As this information becomes more widely available to patients and third-party payors, radiosurgery may replace surgical resection as the preferred management strategy for patients with small to medium sized vestibular schwannomas in the United States. RATIONALE: It is estimated that 2500 patients are diagnosed with vestibular schwannomas each year in the United States. Assuming that 80% undergo surgery, 2000 operations are performed annually for newly diagnosed vestibular schwannomas. Data available since 1987 regarding the number of cases for which gamma knife radiosurgery was performed were used to predict the number of patients who will undergo vestibular schwannoma radiosurgery in the future. If the current trend continues, an equal number of patients will undergo surgical resection and radiosurgery to treat their vestibular schwannomas (approximately 1000/yr) sometime between 2005 and 2010. Moreover, it is predicted that by 2020, two-thirds of the patients who are newly diagnosed with vestibular schwannomas will undergo radiosurgery, with surgical resection being reserved for patients with large tumors associated with symptomatic brain stem compression. DISCUSSION: Early data regarding vestibular schwannoma radiosurgery predicted an exponential growth curve. Although it is premature to assume that the current trend will continue, it is likely that an ever increasing percentage of patients will undergo radiosurgery as accessibility to this alternative increases, and more data are published regarding long-term tumor growth control rates. If the mathematical model proves to be accurate, then stereotactic radiosurgery will replace surgical resection as the preferred management strategy for the majority of patients with vestibular schwannomas. PMID- 9733303 TI - Computed tomography-guided trigeminal tractotomy-nucleotomy in the management of vagoglossopharyngeal and geniculate neuralgias. AB - OBJECTIVE: Vagoglossopharyngeal and geniculate neuralgias are less frequently seen types of cranial neuralgias. Their causes and symptomatology are similar to those of trigeminal neuralgia; however, the complex anatomic relationship between the intermedius, vagal, and glossopharyngeal nerves leads to difficulties in the diagnosis and management of neuralgias originating from these cranial nerves. Numerous procedures have been used to treat intractable neuralgias of the VIIth, IXth, and Xth cranial nerves: 1) extracranial sectioning of the cranial nerves, 2) percutaneous thermal rhizotomy, 3) intracranial glossopharyngeal and vagal rhizotomies, 4) microvascular decompression, and 5) percutaneous trigeminal tractotomy-nucleotomy (TR-NC) or nucleus caudalis dorsal root entry zone operation. We propose that computer-guided TR-NC may be the first-choice operation for patients with glossopharyngeal, vagal, or geniculate neuralgia. PATIENTS AND METHODS: Nine patients suffering from idiopathic vagoglossopharyngeal neuralgia (six patients) and geniculate neuralgia (three patients) were managed at our clinic. Computed tomography-guided percutaneous trigeminal TR-NC was performed for these nine patients. RESULTS: Excellent (six patients) or good (three patients) pain control was obtained in each patient. Complications included temporary ataxia in two patients after TR-NC. CONCLUSION: The risk:benefit ratio should be evaluated individually to select the appropriate treatment procedure for patients with vagoglossopharyngeal and geniculate neuralgias. Computed tomography-guided percutaneous TR-NC is an effective and minimally invasive procedure for such patients. PMID- 9733304 TI - The predictive value of intraoperative somatosensory evoked potential monitoring: review of 244 procedures. AB - INTRODUCTION: There is some controversy regarding the value of intraoperative neurophysiological monitoring in predicting postoperative neurological deficits. We discuss our experience with the use of intraoperative somatosensory evoked potentials (SSEPs) during surgery of cranial base tumors. METHODS: We retrospectively reviewed all of the procedures that had been performed for the resection of cranial base tumors from July 29, 1993, through March 16, 1995. One hundred ninety-three consecutive patients had undergone a total of 244 procedures. SSEP waveforms were classified as follows: Type I, no change; Type II, change that reverts to baseline; Type III, change that does not revert to baseline; and Type IV, complete flattening of the SSEP waveform without improvement. Two patients had no waveforms from the beginning of the case (Type V) and were excluded from further analysis. New immediate postoperative neurological deficits were recorded. RESULTS: There were 64 male and 129 female patients, with a mean age of 46.6 years. One hundred seventy-seven patients had Type I SSEP waveforms, 13 of whom had postoperative deficits (7%). Fifty-six patients had Type II SSEPs, and nine (16%) of them had postoperative neurological deficits. Six patients had Type III SSEPs, and three had Type IV SSEPs, all of whom (100%) had postoperative deficits. There was a correlation between SSEP type and the results of the postoperative neurological examinations. The positive predictive value is 100%, and the negative predictive value is 90%. Although a change in the waveform that did not revert to baseline (Types III and IV) always predicted a postoperative deficit, a normal waveform did not always rule out postoperative deficits. Pathological abnormality, vessel encasement, vessel narrowing, degree of cavernous sinus involvement, brain stem edema, middle fossa location, final amount of resection, age, and tumor size correlated with a high predictive value of SSEP monitoring on univariate analysis (P < 0.05). None of these variables correlated significantly on multivariate analysis (P > 0.05), although brain stem edema was close (P = 0.0571). CONCLUSION: Intraoperative SSEPs have a high positive predictive value during surgery for cranial base tumors, but they do not detect all postoperative deficits. PMID- 9733305 TI - Vascular reconstruction using interposed small vessels. AB - OBJECTIVE: This study presents the relationship between the patency of short vessel graft bypasses and their diameter/length. METHODS: The authors performed interposed graft bypass operations using small vessels for four patients with moyamoya disease, six patients with cerebral thrombosis, and one patient with aortitis syndrome. The donor artery was the superficial temporal artery (10 patients) or the occipital artery (1 patient), and the recipient artery was the cortical branch of the middle cerebral artery (8 patients) or the cortical branch of the anterior cerebral artery (3 patients). The interposed graft used between these donor and recipient vessels was the superficial temporal vein (seven patients), the superficial temporal artery (three patients), or the epigastric artery (one patient). RESULTS: Good patency of the graft was confirmed for 7 of these 11 patients. Regarding the relationship between the diameter/length and the patency, we found that long-term patency could not be expected when the discriminant function of y = (15.39 x diameter) - (0.35 x length) - 14.37 was below zero. CONCLUSION: Short-vessel graft bypass is a practical option for cerebral revascularization surgery when short large vessels are used. PMID- 9733306 TI - Pallidotomy lesion locations: significance of microelectrode refinement. AB - OBJECTIVE: To determine whether stereotactic pallidotomy requires refinement using microelectrode recording to ensure proper lesion placement. METHODS: The experiment approach was based on retrospective comparisons of microelectrode refined radiofrequency lesion locations with hypothetical unrefined lesion positions. Actual and hypothetical pallidotomy lesions were classified based on their lesion center (thermocoagulative zone) locations and their total lesion areas (surrounding edematous zone) relative to the pallidal target. Assessments were made using postoperative T2-weighted magnetic resonance axial images, which showed both the lesion and globus pallidus (GP). The magnitude of microelectrode refinement from an initial preoperative starting point determined by computed tomography was calculated using stereotactic coordinates and included corrections for the lesioning tract trajectory angle. RESULTS: In all 25 patients, the center of the actual pallidotomy lesion was within the GP. Without microelectrode refinement, 13 of 25 hypothetical lesion positions would have been localized such that the lesion center would not have remained in the GP. In eight cases, microelectrode refinement resulted in no significant change in lesion location, but in one case, microelectrode refinement resulted in lesion center placement away from the GP. CONCLUSION: Kinesthetically driven microelectrode refinement in pallidotomy lesioning seems to be required to ensure proper lesion location within the GP. PMID- 9733307 TI - Measurement of intraoperative brain surface deformation under a craniotomy. AB - OBJECTIVE: Several causes of spatial inaccuracies in image-guided surgery have been carefully studied and documented for several systems. These include error in identifying the external features used for registration, geometrical distortion in the preoperative images, and error in tracking the surgical instruments. Another potentially important source of error is brain deformation between the time of imaging and the time of surgery or during surgery. In this study, we measured the deformation of the dura and brain surfaces between the time of imaging and the start of surgical resection for 21 patients. METHODS: All patients underwent intraoperative functional mapping, allowing us to measure brain surface motion at two times that were separated by nearly an hour after opening the dura but before performing resection. The positions of the dura and brain surfaces were recorded and transformed to the coordinate space of a preoperative magnetic resonance image, using the Acustar surgical navigation system (manufactured by Johnson & Johnson Professional, Inc., Randolph, MA) (the Acustar trademark and associated intellectual property rights are now owned by Picker International, Highland Heights, OH). This system performs image registration with bone-implanted markers and tracks a surgical probe by optical triangulation. RESULTS: The mean displacements of the dura and the first and second brain surfaces were 1.2, 4.4, and 5.6 mm, respectively, with corresponding mean volume reductions under the craniotomy of 6, 22, and 29 cc. The maximum displacement was greater than 10 mm in approximately one-third of the patients for the first brain surface measurement and one-half of the patients for the second. In all cases, the direction of brain shift corresponded to a "sinking" of the brain intraoperatively, compared with its preoperative position. Analysis of the measurement error revealed that its magnitude was approximately 1 to 2 mm. We observed two different patterns of the brain surface deformation field, depending on the inclination of the craniotomy with respect to gravity. Separate measurements of brain deformation within the closed cranium caused by changes in patient head orientation with respect to gravity suggested that less than 1 mm of the brain shift recorded intraoperatively could have resulted from the change in patient orientation between the time of imaging and the time of surgery. CONCLUSION: These results suggest that intraoperative brain deformation is an important source of error that needs to be considered when using surgical navigation systems. PMID- 9733308 TI - Virtual endoscopy for planning and simulation of minimally invasive neurosurgery. AB - OBJECTIVE: This article demonstrates the usefulness and the problems of present state software for virtual endoscopy as a tool for the planning and simulation of minimally invasive neurosurgical procedures. METHODS: The software Navigator (General Electric Medical Systems, Buc, France) was applied for virtual endoscopic visualization of three-dimensional magnetic resonance data sets of healthy volunteers and neurosurgical patients, using a clinical magnetic resonance scanner (1.5-T Signa Hispeed; General Electric Medical Systems). Classical approaches for minimally invasive procedures were simulated. RESULTS: Virtual endoscopy provided impressive three-dimensional views of intracranial and intracerebral cavities, with visualization of many anatomic details of the brain's inner and outer surfaces. The method proved to be especially suited for the simulation and planning of operations of intraventricular lesions, for which the technical limitations of the present state of development of this method have fewer implications. However, the present state of technology, as described in this article, has two major shortcomings: 1) the blood vessels cannot be visualized together with the brain tissue and cranial nerves; and 2) different tissue compartments cannot be stained in their original coloring, which would facilitate their recognition and thus orientation in space by anatomic landmarks. Another important disadvantage at this stage is time consumption for many single working steps. CONCLUSION: Virtual endoscopy is a promising tool for teaching and training in intracranial neuroanatomy as well as for planning and simulation of minimally invasive (e.g., endoscopic), mainly intraventricular, operations. Direct clinical application is, at this stage of development, limited by several technical shortcomings of visualization and quantification of distances and modeling of surfaces. PMID- 9733309 TI - The role of angioplasty in the treatment of cerebrovascular disease. AB - Percutaneous transluminal angioplasty (PTA), an established treatment of arterial stenosis in coronary, renal, and other peripheral sites, is being applied to the cerebrovascular territory with greater frequency. Early results suggest that PTA may be safe and efficacious in the treatment of extracranial arterial stenosis secondary to atherosclerosis and fibromuscular dysplasia. PTA is also being used with promising results in treating symptomatic intracranial arterial stenosis from atherosclerosis. This review examines PTA in the treatment of cerebrovascular disease, current indications, and results. PMID- 9733310 TI - Persistent trigeminal artery: an anatomic study. AB - INTRODUCTION: The most frequent embryonic communication between the vertebrobasilar and carotid systems is a persistent trigeminal artery (PTA). It has been observed in 0.1 to 0.2% of cerebral angiograms. We found this variation in an anatomic specimen, and after microscopic dissection, we performed an analysis of the course of the PTA and its relationship with the abducens nerve and the meningohypophyseal trunk. METHOD: A PTA was incidentally encountered in an injected cadaver specimen during a transpetrosal approach. This embryonic variation and its anatomic relationship are discussed. RESULTS: The PTA can take either a lateral or medial course regarding its relationship with the abducens nerve. When the PTA originates from the posterolateral aspect of the posterior bend of the cavernous carotid artery (C4 segment), it crosses underneath and distorts the abducens nerve, continuing between the abducens and trigeminal nerves. When taking a medial course, the PTA arises from the posteromedial aspect of the posterior bend of the cavernous carotid at the same segment and pierces the clival dura at the dorsum sellae. Cranial nerve displacement or distortion is less likely in this variation. In an analysis of carefully described anatomic studies, the PTA and meningohypophyseal trunk were found arising from either common or separated origins. CONCLUSION: The most frequent embryological anastomosis between the carotid and vertebrobasilar system is the PTA. Its course and relationship with the cranial nerves may determine its clinical presentation. PMID- 9733311 TI - Anatomy of the frontotemporal branch of the facial nerve and indications for interfascial dissection. AB - INTRODUCTION: Many studies have been conducted of the surgical anatomy of the frontotemporal branch of the facial nerve (FTBFN). However, very few have addressed the indications for interfascial dissection. When the zygomatic arch needs to be exposed, the interfascial approach is recommended to protect the FTBFN. With the transbasal or subfrontal approaches, however, when a bicoronal skin incision is used, the need for the interfascial approach is not clear. METHODS: We studied 10 temporal regions (5 cadaveric heads). We dissected the recognized fascial layers of the temporal region and the FTBFN. We performed a histological study in a sixth specimen. RESULTS: We observed the following. 1) The galea and the superficial layer of the deep temporal fascia become fused in a curved line from the lateral orbital border 2.8 cm above the zygomatic arch to a point 3 cm posterior to the inferolateral angle of the orbit. 2) After this transitional area of adherence, the subgaleal loose cellular layer is lost and is replaced by a fibrofatty tissue. 3) The FTBFN in its course above the zygomatic arch runs in this tissue layer without being protected by the galea. 4) Over the superolateral angle of the orbital rim, the galea protects FTBFN, and there are no subgaleal adhesions in that area. CONCLUSION: Ahove the zygomatic arch, the FTBFN is not protected by the galea. During bicoronal approaches, if only the superolateral angle of the orbital rim needs to be exposed and not the zygomatic arch, there is no need to protect the FTBFN using an interfascial approach. PMID- 9733312 TI - Delayed vascular changes after antiangiogenic therapy with antivascular endothelial growth factor antibodies in human glioma xenografts in nude mice. AB - OBJECTIVE: The purpose of this study was to examine the delayed effects of antivascular endothelial growth factor treatment on tumor growth and vascularity in a subcutaneous mouse tumor model of human glioblastoma. METHODS: Antivascular endothelial growth factor antibody treatment was administered for a period of 6 weeks, to suppress tumor growth. To detect late vascular effects, tumor vascular parameters for treated tumors and control tumors were analyzed 4 weeks thereafter. By that time, tumors had grown to adequate sizes (diameter, 8-10 mm) for comparison with untreated control tumors. Vascular parameters were quantified by using an image-analysis system. RESULTS: Vascular density was significantly lower in antivascular endothelial growth factor antibody-treated tumors, compared with control tumors of similar size. The vascular architecture of treated tumors was also distinctly different, compared with control tumors, showing larger but sparser vessel structures. CONCLUSION: These findings suggest that antiangiogenic therapy may have a prolonged effect on the vascular architecture of certain tumors, resulting in enduring changes in the tumor vessels. Because tumor vasculature plays an important role in the sensitivity to various treatment modalities, these changes are likely to influence the responses of these tumors to further therapy. PMID- 9733313 TI - Reactive oxygen species in reoxygenation injury of rat brain capillary endothelial cells. AB - OBJECTIVE: To clarify the mechanism of anoxia/reoxygenation (A/R) injury of rat brain capillary endothelial cells (BCEC). METHODS: BCEC isolated from Sprague Dawley rats by enzymatic treatment and centrifugation were subjected to anoxia (95% N2, 5% CO2) for 20 minutes and then to reoxygenation (95% air, 5% CO2) for 3 hours. Enzyme inhibitors, including oxypurinol, indomethacin, and N(G)-nitro-L arginine methyl ester, or specific free-radical scavengers, such as superoxide dismutase, catalase, and the ferric iron chelator deferoxamine, were added before A/R injury. The BCEC were incubated in a range of Ca2+ concentrations from 1 to 0.01 mmol/L during A/R injury. Cytotoxicity was assayed by release of intracellular lactate dehydrogenase (LDH). RESULTS: With A/R injury, LDH release from the control group (no protective agents) significantly increased (44.8 +/- 3.3%), compared with a small increase in a normoxic group. BCEC treated with oxypurinol, indomethacin, or N(G)-nitro-L-arginine methyl ester showed suppression of LDH release. LDH release was almost totally suppressed by superoxide dismutase and partially by catalase or deferoxamine. The LDH release was partly dependent on calcium concentration. CONCLUSION: BCEC subjected to A/R become potent generators of free radicals, especially superoxide anion. Free radical production depends on both xanthine oxidase and cyclooxygenase pathways. Peroxynitrite and extracellular Ca2+ both contribute importantly to reoxygenation injury of BCEC. PMID- 9733315 TI - Saccular aneurysm induction by elastase digestion of the arterial wall: a new animal model. AB - OBJECTIVE: To develop a rabbit aneurysm model that is more realistic in gross appearance and histological features than previous models and to enable the development of a larger animal model. METHODS: Ten rabbits received porcine pancreatic elastase, five at the right common carotid artery bifurcation and five others at the right superior thyroid artery origin. One control animal received collagenase and another received papaverine, each at the right superior thyroid artery origin. The agents were topically delivered to the arterial adventitia with a microsyringe after surgical exposure of the targeted arteries. The arteries were monitored for aneurysm growth with a video camera for up to 3 hours and were then removed and processed for histology. RESULTS: Saccular aneurysms developed in one of five animals after elastase application at the carotid bifurcation and in all five animals receiving elastase at the superior thyroid artery origin. Among the six aneurysms, recurrent minor hemorrhages occurred in four, thrombosis of the aneurysm sac in three, and rupture causing severe bleeding in one. Histological sections revealed thin-walled aneurysms composed only of collagen fibers and some cellular elements. No saccular dilation resulted from papaverine application. Collagenase application resulted in a hemorrhagic thrombotic lesion in the arterial wall but no aneurysm formation. CONCLUSION: Arterial saccular aneurysms were induced in rabbits by topical application of elastase with an easy and efficient method. These aneurysms are histologically similar to natural aneurysms, and their arterial nature renders them more authentic than those of surgical models. This aneurysm model may serve as a foundation for further aneurysm research. PMID- 9733314 TI - Posttraumatic cerebral ischemia after fluid percussion brain injury: an autoradiographic and histopathological study in rats. AB - OBJECTIVES: Mild-to-moderate reductions in local cerebral blood flow (ICBF) have been reported to occur in rats after moderate (1.7-2.2 atm) fluid percussion brain injury. The purpose of this study was to determine whether evidence for severe ischemia (i.e., mean ICBF < 0.25 ml/g/min) could be demonstrated after severe brain injury. In addition, patterns of indium-labeled platelet accumulation and histopathological outcome were correlated with the hemodynamic alterations. METHODS: Sprague-Dawley rats (n = 23), anesthetized with halothane and maintained on a 70:30 mixture of nitrous oxide:oxygen and 0.5% halothane, underwent normothermic (37 degrees C) parasagittal fluid percussion brain injury (2.4-2.6 atm). Indium-111-tropolone-labeled platelets were injected 30 minutes before traumatic brain injury (TBI), while 14C-iodoantipyrine was infused 30 minutes after trauma for ICBF determination. Sham-operated animals (n = 8) underwent similar surgical procedures but were not injured. For histopathological analysis, traumatized rats (n = 5) were perfusion-fixed 3 days after TBI. RESULTS: In autoradiographic images of indium-labeled platelets, abnormal platelet accumulation that was most pronounced overlying the pial surface was commonly associated with severe reductions in ICBF within underlying cortical regions 30 minutes after TBI. For example, within the lateral parietal cortex, ICBF was significantly reduced from 1.67 +/- 0.11 ml/g per minute (mean +/- standard error of the mean) in sham-operated animals to 0.23 +/- 0.03 ml/g per minute within the traumatized group. In addition to focal severe ischemia, moderate reductions in ICBF were detected throughout the traumatized hemisphere, including the frontal and occipital cortices, hippocampus, thalamus, and striatum. Mild decreases in ICBF were also observed throughout the contralateral cerebral cortex. At 3 days after severe TBI, histopathology demonstrated intracerebral and subarachnoid hemorrhage associated with cerebral contusion and selective neuronal necrosis. CONCLUSION: These data indicate that multiple cerebrovascular abnormalities, including subarachnoid hemorrhage, focal platelet accumulation, and severe ischemia, are important early events in the pathogenesis of cortical contusion formation after TBI. Injury severity is expected to be a critical factor in determining what therapeutic strategies are attempted in the clinical setting. PMID- 9733316 TI - The mystery of angiography and the "unawarded" Nobel Prize: Egas Moniz and Hans Christian Jacobaeus. AB - OBJECTIVE: To investigate the circumstances surrounding why Egas Moniz was not awarded the Nobel Prize for his contribution of angiography, provide a synopsis of Moniz's political and medical careers, and present a biographical sketch of Hans Christian Jacobaeus, the neurologist who evaluated Moniz's Nobel Prize nominations, as well as to dispel long-standing misconceptions concerning Moniz's recognition and to acknowledge the contributions of other researchers. HISTORICAL PERSPECTIVE: In 1936, Antonio Caetano de Abreu Freire Egas Moniz published the results of a radical treatment for mental illness, a surgical procedure he termed prefrontal leucotomy. Moniz achieved such remarkable results with mental patients who suffered from conditions previously deemed incurable that many physicians throughout the world immediately embraced the procedure. In 1949, the Nobel Prize Committee recognized Moniz's contribution with the Nobel Prize in Physiology and Medicine. Why Moniz's earlier major contribution to medicine, the discovery and development of angiography, was not acknowledged in like fashion has remained a mystery. Nobel Prize documents reveal that Moniz was nominated for the award on two separate occasions; both times, Jacobaeus, Chairman of the Department of Neurology at Karolinska Institute and a member of the Nobel Prize Committee, evaluated the nominations and recommended against awarding Moniz the prize. CONCLUSION: The development of imaging techniques was not isolated to any one individual's contribution. Several persons, including Walter Dandy and Jacobaeus, were leading figures. PMID- 9733317 TI - Atypical central nervous system lymphoma at the cranial base: report of four cases. AB - OBJECTIVE AND IMPORTANCE: Primary central nervous system lymphoma is a disease with increasing incidence. Atypical presentations are becoming more frequent. CLINICAL PRESENTATION: Three patients bearing cavernous sinus lesions presented initially with periorbital pain and diplopia. Tolosa-Hunt syndrome was the initial presumptive diagnosis for two patients, and meningioma was the diagnosis for the third patient. A fourth patient presented with left ear pain, and a mass in the left internal auditory canal was thought to represent an acoustic neuroma. INTERVENTION: For all four patients, an operative pathological diagnosis was obtained and was compatible with central nervous system lymphoma. All patients were treated with osmotic blood-brain barrier disruption with intra-arterial chemotherapy using a methotrexate-based regimen. CONCLUSION: All four cases included atypical presentations of lymphoma. These cases again illustrate that a correct diagnosis cannot be obtained based only on imaging and clinical findings. PMID- 9733319 TI - Trigeminal neuralgia resulting from infarction of the root entry zone of the trigeminal nerve: case report. AB - OBJECTIVE AND CLINICAL IMPORTANCE: We present a case of trigeminal neuralgia resulting from infarction of the root entry zone of the trigeminal nerve. This is the first reported case of an unusual cause of trigeminal neuralgia. CLINICAL PRESENTATION: A 71-year-old man presented with severe lancinating pain in the left V1 and V2 distributions. Magnetic resonance imaging of the brain demonstrated a small wedge-shaped infarct at the root entry zone of the left trigeminal nerve in the pons. INTERVENTION: Medical management with carbamazepine was initially successful, but the patient later developed refractory pain and was unable to tolerate side effects of the medication. The patient underwent subsequent percutaneous glycerol rhizotomy, which resulted in complete resolution of his pain. CONCLUSION: Infarction of the root entry zone may produce typical symptoms of trigeminal neuralgia similar to a multiple sclerosis plaque at the root entry zone. Treatment of trigeminal neuralgia must consider the underlying cause. Glycerol rhizotomy may provide relief of pain for patients in whom there is no evidence of vascular compression. PMID- 9733318 TI - Abscess formation in invasive pituitary adenoma: case report. AB - OBJECTIVE AND IMPORTANCE: Pituitary abscess is rare, with few cases of secondary pituitary abscess reported. To our knowledge, only 14 cases of pituitary adenomas with abscess have been reported, and this is the first report to include a magnetic resonance image of a pituitary adenoma with abscess. CLINICAL PRESENTATION: A 22-year-old man presented with decreasing vision, headache, and vomiting. His higher functions were normal, with visual disturbance in both eyes. Computed tomographic scans and magnetic resonance images were suggestive of a sellar tumor with a cystic component. INTERVENTION: A right pterional craniotomy was performed, and the tumor was debulked, revealing an abscess in the center. Postoperatively, the patient received a regimen of antibiotics. CONCLUSION: Early surgical intervention plus follow-up antibiotic therapy are the mainstay of treatment for abscess formation in pituitary adenomas. PMID- 9733320 TI - Abducens nerve palsy after radiofrequency rhizolysis for trigeminal neuralgia: case report. AB - OBJECTIVE AND IMPORTANCE: Abducens nerve palsy is a rare and reversible complication associated with percutaneous radiofrequency trigeminal rhizolysis. CLINICAL PRESENTATION: An 86-year-old man developed an abducens nerve palsy immediately after undergoing percutaneous radiofrequency trigeminal rhizolysis for severe trigeminal neuralgia involving all three divisions of the trigeminal nerve. The palsy resolved spontaneously after 2 months. CONCLUSION: This case, in combination with previous reports, suggests that cases of transient sixth nerve palsy associated with percutaneous radiofrequency trigeminal rhizolysis for trigeminal neuralgia are more likely to occur in elderly patients with ophthalmic division involvement. PMID- 9733321 TI - Ruptured de novo aneurysm induced by ethyl 2-cyanoacrylate: case report. AB - OBJECTIVE AND IMPORTANCE: We report a rare case of a ruptured de novo aneurysm induced by ethyl 2-cyanoacrylate. CLINICAL PRESENTATION: A 44-year-old woman had undergone microvascular decompression for a right-sided facial spasm. The preoperative vertebral angiogram did not show any aneurysmal dilation. The right anteroinferior cerebellar artery, which was compressing the exit zone of the facial nerve, was detached and fixed to the dura mater with ethyl 2 cyanoacrylate. Nine years later, the patient suffered a subarachnoid hemorrhage caused by the rupture of a newly developed aneurysm of the right anteroinferior cerebellar artery. INTERVENTION: The aneurysm was clipped 2 days after onset of the subarachnoid hemorrhage. It consisted of two bulges in the arterial wall on the proximal side of the meatal loop. One bulge was stuck to the dura mater of the pyramis by ethyl 2-cyanoacrylate, which had been used in the microvascular decompression 9 years previously. CONCLUSION: This is the first reported clinical case of a de novo aneurysm induced by a cyanoacrylate adhesive. Ethyl 2 cyanoacrylate can damage the arterial wall and induce a de novo aneurysm. PMID- 9733323 TI - Carotid artery balloon test occlusion: combined clinical evaluation and xenon enhanced computed tomographic cerebral blood flow evaluation without patient transfer or balloon reinflation: technical note. AB - OBJECTIVE: Clinical evaluation was combined with xenon-enhanced computed tomographic (CT) cerebral blood flow (CBF) evaluation during carotid artery balloon test occlusion (BTO), without patient transfer from the angiography suite to the CT scanner or balloon reinflation. TECHNIQUE: Thirteen patients underwent carotid artery BTO. Placement of temporary occlusion balloons was performed with patients positioned on the CT scanner table. If neurological testing revealed no changes within 10 minutes after balloon inflation, patients were positioned within the CT scanner gantry for xenon-enhanced CT CBF evaluation. CBF evaluations were begun 12 to 15 minutes after balloon inflation and required 8 minutes for completion. After completion of CBF evaluation, neurological testing continued during 30 minutes of arterial occlusion. RESULTS: One patient did not tolerate BTO, with the development of reversible hemiparesis. Reliable CBF data were not obtained because of patient motion in one case. Eleven patients clinically tolerated BTO and completed CBF evaluation. For five patients, xenon enhanced CT scanning revealed no regions with CBF of less than 30 ml/100 g/min. For four patients, xenon-enhanced CT scanning revealed small regions with CBF of less than 30 ml/100 g/min within the anterior frontal lobe on the occluded side. For two patients, ipsilateral CBF decreased dramatically during BTO, with CBF in many regions of less than 30 ml/100 g/min and in some of less than 20 ml/100 g/min. CONCLUSION: Xenon-enhanced CT CBF evaluation can be combined with clinical testing during BTO without patient transfer, balloon reinflation, or increases in the duration of the procedure. We recognize that the value of CBF evaluation during BTO remains to be proven; our technique does, however, eliminate abbreviated clinical neurological evaluation, patient transfer, and balloon reinflation, which were previously associated with the use of xenon-enhanced CT CBF evaluation during carotid artery BTO. PMID- 9733322 TI - Cervical myelopathy caused by hypoplasia of the atlas: two case reports and review of the literature. AB - OBJECTIVE AND IMPORTANCE: Congenital anomalies of the posterior arch of the atlas (C1) are uncommon. They range from partial clefts to total agenesis of the posterior arch. Developmental cervical canal stenosis is a congenital anomaly that may cause cervical myelopathy. Myelopathy caused by cervical stenosis at the level of the atlas has been reported in only three cases. We present two cases of nontraumatic cervical myelopathy caused by spinal stenosis at the level of the atlas associated with a hypoplastic but complete posterior arch of C1. CLINICAL PRESENTATION: Two elderly Chinese men developed cervical myelopathy gradually during months to years, without preceding trauma. Imaging revealed a hypoplastic but complete posterior C1 arch associated with changes of spondylosis in both patients, producing severe spinal stenosis and spinal cord compression. Posterior decompression was achieved in both by the removal of the posterior arch of C1 with its surrounding thickened posterior ligaments. Symptoms and clinical findings improved in the two patients during the follow-up period. CONCLUSION: The anomaly presented in our two cases differs from the established classification of congenital abnormalities of the posterior arch of the atlas, suggesting a different embryological defect. The hypoplastic posterior C1 arch created a congenitally narrowed spinal canal in our patients, rendering the spinal cord more susceptible to compression related to degenerative changes of the spine. Surgical removal of the shortened posterior C1 arch and surrounding degenerative ligaments is an effective treatment for symptomatic patients with this condition. PMID- 9733324 TI - Functional magnetic resonance imaging localization of ictal onset to a dysplastic cleft with simultaneous sensorimotor mapping: intraoperative electrophysiological confirmation and postoperative follow-up: technical note. AB - INTRODUCTION: Although technically challenging to obtain, ictal functional magnetic resonance imaging has been used to localize ictal onset zones in a small number of patients. We used this technique to demonstrate the inherent epileptogenicity of dysplastic cortex. METHODS: We present a 16-year-old female patient with intractable left-sided sensorimotor seizures and a congenital dysplastic cleft lying along the right rolandic fissure. Preoperative functional magnetic resonance imaging (blood oxygen level-dependent sequence, 1.5 T) localized the motor and sensory cortices to the anterior border of the cleft. During a speech activation run, the patient experienced a 20-second seizure. Initial activation was seen within the dysplastic cortex along the deep posterior margin of the cleft. Intraoperative median nerve stimulation produced a distinct N20/P20 wave inversion over the dysplastic cleft. Stimulation mapping performed with the patient awake confirmed the location of the sensorimotor cortex on the anterior border of the cleft, and preresection electrocorticography identified abundant interictal spikes along the posterior border after opening the cleft. RESULTS: After surgical resection of the dysplastic cortex, the patient exhibited transient minimal weakness and mild neglect, which resolved within 1 week. Two years after surgery, she was neurologically intact and seizure-free. CONCLUSION: This study used functional magnetic resonance imaging to demonstrate the inherent epileptogenicity of dysplastic cortex and to simultaneously map ictal and functional cortex. The N20 wave inversion can be a useful intraoperative tool for identifying the central sulcus (or its equivalent), even in the presence of abnormal cortical architecture. PMID- 9733325 TI - Evaluation of cerebral vasospasm after early surgical and endovascular treatment of ruptured intracranial aneurysms. PMID- 9733326 TI - Traumatic basilar aneurysm after endoscopic third ventriculostomy: case report. PMID- 9733327 TI - Intracerebral inflammation after human brain contusion. PMID- 9733328 TI - Intramedullary pressure in syringomyelia: clinical and pathophysiological correlates of syrinx distension. PMID- 9733329 TI - Acute posttraumatic subdural hematomas: "intradural" computed tomographic appearance as a favorable prognostic factor. PMID- 9733331 TI - Tacrine efficacy in Lewy body dementia. AB - BACKGROUND: Response to tacrine varies among patients with Alzheimer's disease (AD). Lewy body dementia (LBD) could be a high responder subtype of AD. The aim of the study was to compare the effects of tacrine in LBD and AD. METHODS: Seventy-five consecutive outpatients with mild or moderate AD were screened. Tacrine was given at a dose of 40 mg/day during 6 weeks. During the next 6 weeks, the patients were treated with 80 mg/day and afterwards with 120 mg/day. Patients were assessed at baseline and treated with a dose of 120 mg/day tacrine for 2 weeks. RESULTS: Analysis was performed on 39 patients (AD, N = 20; LBD, N = 19). Eight patients were lost to follow-up, eight patients manifested with side effects, six suffered from an intercurrent somatic disease during the study and 14 patients had poor compliance or were treated with incompatible drugs. Twenty two patients (11 AD/11 LBD) increased their cognitive performances with tacrine. Among the 22 patients, the improvement differed between the AD and the LBD groups. In AD, conceptualization improved; in LBD, the improvements occurred in verbal initiation and digit span. CONCLUSION: This study emphasizes the importance of using appropriate tests to determine the positive effects of pharmacological treatments. PMID- 9733330 TI - Adjustment to residential placement in Alzheimer disease patients: does premorbid personality matter? AB - AIM: To evaluate the influence of premorbid personality on adaptation to placement in a long-term care facility. SUBJECTS: Twenty-eight persons with probable Alzheimer disease (AD) residing in an academically affiliated nursing home for 6-9 months. METHODS: Premorbid personality was described retrospectively by two informants for each resident using the revised NEO Personality Inventory (NEO-PI-R). Standardized tests and rating scales were used on admission to the facility to assess cognition, mood state, physical dependency and general health. Nurses rated each AD resident's social behaviour, participation in activities and quality of sleep. RESULTS: Poorer adjustment was associated with more severe dementia but better physical health. None of the NEO-PI-R domain scores predicted adjustment. CONCLUSIONS: Contrary to popular belief, premorbid personality is relatively inconsequential for an AD patient's adaptation to a long-term care facility. PMID- 9733332 TI - Burnout: current knowledge and relevance to old age psychiatry. AB - OBJECTIVES: To review the literature on burnout and consider its relevance to old age psychiatry and the role of the consultant. DATA SOURCES: Medline and PsychLit computerized databases. DATA SYNTHESIS: Burnout is a syndrome of emotional exhaustion, depersonalization and decreased sense of personal accomplishment which is recognized in people working in the human service professions and can have adverse effects on the workforce. There is little evidence of unique stressors related to care of elderly mentally ill people. Burnout is likely to be modified by workplace interventions. Relevant areas for intervention are political and social, organizational and management, training and personal issues. Support to consultants and their continuing professional development need to be radically reviewed. PMID- 9733333 TI - Early loss of mother or father predicts depression in old age. AB - The independent predictive roles of early losses, personality traits, acute losses and long-term stress situations for the occurrence of depression in elderly Finns were described using a longitudinal design. The persons non depressed in an epidemiological study in 1984-85 were interviewed in 1989-90 (N = 679) and the occurrence of depression was determined according to DSM III criteria. Logistic regression models were used to assess the independent roles of the hypothesized factors as predictors. An early loss of the mother among men and an early loss of the father among women independently predicted the occurrence of depression in logistic regression models. Older age in men, and a higher number of symptoms, the occurrence of previous depression and not living alone in women were also independent predictors. In men, impaired functional abilities and poor self-perceived health tended to predict depression. In conclusion, the psychological trauma which develops upon the experience of an early parental loss contributes to the development of depression even in old age. The role of stressors in life as independent predictors of depression in old age was also ascertained. PMID- 9733334 TI - The economic consequences of Alzheimer's disease in the context of new drug developments. AB - The first national symptomatic treatment for Alzheimer's disease has received a very mixed and perhaps ageist reception from purchasers of health care in the UK. This is largely because detailed information on the long-term effects of this class of drugs is scarce. However, by looking at the published evidence on the economic burden of Alzheimer's disease, some observations and assumptions can be made as to the influence of the new drug treatments. The drug therapies available and those most likely to become licensed are reviewed and the potential economic impact is discussed. Long-term outcome studies would properly address this, but as these drugs have now demonstrated efficacy, particularly in non-cognitive behaviours, it will be ethically more difficult to maintain patients on placebo for long periods. Some assumptions therefore have to be made from long-term open label studies. Those drugs currently available, and those in development, may offer effective treatment for some of the core symptoms of Alzheimer's disease, slowing the rate of cognitive decline and preserving competence in activities of daily living for longer. If handled correctly, these treatments have the potential to offer cost savings for many patients, and cost-effectiveness improvements look probable. PMID- 9733335 TI - Severity of cognitive impairment in Alzheimer's disease affects list learning using the California Verbal Learning Test (CVLT). AB - Impairment in list learning is considered a primary symptom of Alzheimer's disease (AD), yet there are no published reports examining the relationship between list learning and severity of cognitive impairment. We gave nine-item and 16-item versions of the California Verbal Learning Test (CVLT; Delis et al., 1987), a standardized shopping list assessment of memory, to 24 AD patients (mean age = 76.2 +/- 8.1; mean years of education = 13.8 +/- 2.4), who were stratified into four groups based on MMSE scores (mean = 16.0 +/- 5.6). ANOVAs revealed severity effects for total list learning (p < 0.001), the first trial (p < 0.001), the last trial (p < 0.001) and short- and long-delay recall measures. Most of these differences seemed due to floor effects. For example, the modal number of words recalled after a delay was 0 by subjects with MMSE scores below 21. Severity of cognitive impairment was associated with the proportion of intrusions such that the most severely demented subjects gave almost entirely intrusion responses. Surprisingly, list length did not significantly affect any of the free recall measures. Our results suggest that list learning and recall seem to be lost relatively early in AD. Measures of list recall like the CVLT may not be useful in tracking severity of cognitive impairment over time. PMID- 9733336 TI - Dementia care needs in an area population: case register data and morbidity survey estimates. AB - OBJECTIVE: To compare case register data on the frequency and distribution of known dementia cases in a metropolitan area population with expected total numbers computed from a national disability survey. METHOD: Known cases were enumerated by a cross-sectional census of the Camberwell Dementia Register. Expected total numbers were calculated using the Cognitive Disability (CD) Planning Model, based on the OPCS national survey of disability, 1985-86. RESULTS: Cases ascertained by the Dementia Register census comprised one-fifth of expected total prevalence. The proportion of such cases was higher for persons in long-stay care (1 in 3) than for those in private households (1 in 7). According to the CD Planning Model, cases known to specialist agencies were on average no more severely disabled and dependent than those who were unknown. In terms of absolute numbers, the district nursing and home help services appeared to be the most important untapped sources of case detection, but other research indicates that general practice contacts (not included in the planning model) may be at least equally important. CONCLUSIONS: At any given time, a high proportion of dementia cases, whether in long-stay care or in the community, will be outside the purview of specialist services. Primary care agencies are a major potential source, and a systematic health screening of persons aged over 75 years could be used to realize this potential. PMID- 9733337 TI - The feasibility of electronic tracking devices in dementia: a telephone survey and case series. AB - BACKGROUND: Patients with dementia who go out unaccompanied are at risk of accidents or getting lost. It is not known whether they could benefit from electronic tracking devices or whether such devices are practically feasible. METHOD: The likely demand for an electronic tracking device was assessed by means of a telephone survey of a convenience sample of 99 carers. The practical feasibility of a tracking system was assessed in 24 patients with dementia. RESULTS: The telephone survey suggested that 20% of patients were at continuing risk of traffic accidents and 45% were at continuing risk of getting lost. About 7% could have benefited from using the device at the time of survey and a further 11% could have benefited at an earlier point in their illness. In the feasibility study, only nine patients consistently used the device. In two patients, it was successfully used in a search. One patient was injured by a passing vehicle when he had got lost out of range of the device. A major barrier to using the device was recognizing the risk of getting lost before it happened. CONCLUSION: Significant numbers of patients are at risk. Electronic tracking devices may occasionally be useful in carefully selected cases. PMID- 9733338 TI - Somatization in young versus older female panic disorder patients. AB - BACKGROUND AND RATIONALE: Studies in younger patients with panic disorder suggest greater somatization compared to similarly aged normal controls. Thus, we compared the degree of somatization in young versus older female patients with panic disorder to ascertain whether similarly high levels of somatization exist in older panic disorder patients. METHOD: Community-dwelling subjects were recruited for clinical trials for panic disorder and met Diagnostic and Statistical Manual of Mental Disorders (DSM-III-R) criteria for panic disorder as a primary diagnosis. Our sample (N = 64) contained 42 younger females (< 55 years of age; age range 21-54, mean age 34.6) who were compared to 22 older females (> or = 55 years of age; age range 55-73, mean age 60.8). Subjects were evaluated at baseline using the Self-Report Inventory for Somatic Symptoms (SISS). Statistical analysis of total somatization disorder scores (TSDS) was accomplished by t-tests for independent groups. RESULTS: Older patients showed statistically significantly higher total somatization disorder scores (TSDS) (X = 11.54, SD = 7.45) than did younger patients (X = 8.07, SD = 4.77; t(62) = 2.27, p = < 0.05). CONCLUSION: Our results are suggestive of a higher degree of somatization in older compared to younger female panic disorder patients. PMID- 9733340 TI - A 10-item Hamilton Depression Rating Scale to measure major depressive episode severity in outpatients. PMID- 9733339 TI - Resettlement and the Health of the Nation Outcome Scales (HoNOS) in the elderly. PMID- 9733341 TI - Dementia care in Japan: insurance for long-term care legislation in Japan. PMID- 9733343 TI - Current awareness in geriatric psychiatry. PMID- 9733342 TI - Comparison of the Global Assessment of Functioning scale and the Montgomery Asberg Depression Rating Scale in elderly depressed patients. PMID- 9733344 TI - Pharmacologic preconditioning with monophosphoryl lipid A is abolished by 5 hydroxydecanoate, a specific inhibitor of the K(ATP) channel. AB - We sought to determine the role of opening of adenosine triphosphate (ATP) sensitive potassium channel (K(ATP) channel) in monophosphoryl lipid A (MLA) induced myocardial protection after ischemia/reperfusion (I/R) in rabbit. We used 5-hydroxydecanoate (5-HD), an ischemia-selective inhibitor of K(ATP) channel, to block MLA-stimulated cardiac protection. Four groups of rabbits were studied: group I, MLA-vehicle; group II, MLA; group III, MLA + 5-HD; and group IV, 5-HD only. MLA (35 microg/kg, i.v.) or vehicle were given 24 h before I/R. 5-HD (5 mg/kg) was given 15 min before ischemia. All rabbits underwent 30-min coronary occlusion, followed by 3-h reperfusion. Area at risk was delineated by injection of Evan's blue, and infarct size was determined by tetrazolium staining. Pretreatment with MLA reduced infarct size (percentage of area at risk) from 40+/ 8.6% to 15.1+/-1.5%. The infarct size increased to 51.9+/-5.8% with 5-HD in MLA treated rabbits. 5-HD did not alter infarct size significantly when given in vehicle-treated control rabbits. These data suggest that MLA exerts its protective effect through activation of K(ATP) channel. PMID- 9733346 TI - Endogenous nitric oxide influences arteriovenous anastomosis adrenergic tone in the conscious rabbit ear. AB - The role of nitric oxide (NO) in the control of arteriovenous anastomoses (AVAs) has not been studied in vivo in a thermoregulatory end organ. In this study, the effect of local inhibition of NO synthesis by NG-nitro-L-arginine methyl ester (L NAME) on the microvasculature in the rabbit ear (n=12) was observed in vivo through a chronically implanted ear microvascular chamber. Ear cutaneous blood perfusion (CBP), total auricular arterial flow (TAF), and ear temperature were monitored simultaneously with the direct microvascular observations. Results revealed that intrafacial artery infusion of L-NAME produced significant vasoconstriction of arterioles, AVAs, and venules (p < 0.05). A decrease of ear blood perfusion also was demonstrated by changes of CBP, TAF, and surface temperature. The data provide evidence that basal generation of NO influences the vascular resistance in the thermoregulatory end organ. Moreover, endogenous NO production may be more important in regulating the AVA flow than is flow in other parts of the rabbit ear microvasculature. The effects of NO inhibition on ear microvasculature were not abolished by superior cervical ganglionectomy, indicating that NO production in the rabbit ear is not a neurally mediated mechanism. Further study with a short-term rabbit ear preparation showed that inhibition of NO production with L-NAME enhanced microvascular constrictive responses to extraluminal application of norepinephrine. NO thus appears to play a role of basal vasodilator in opposition to the basal adrenergic vasoconstrictor tone in the rabbit ear. PMID- 9733345 TI - Transition metal chelators reduce directly measured myocardial free radical production during reperfusion. AB - Transition metals such as iron and copper are present in the myocardium and can act as catalysts for the formation of oxygen free radicals during reperfusion after myocardial ischemia. Previous studies suggested that transition metal chelators such as desferrioxamine reduce the production of such radicals and may thereby attenuate postischemic myocardial dysfunction. These studies used spin trapping agents, commonly nitrone compounds, which may themselves influence the severity of the ischemia and reperfusion events being studied. We evaluated two transition metal chelators, desferrioxamine, an iron chelator, and bathocuproine, a copper chelator, by using a new electron paramagnetic resonance technique that does not require the administration of spin traps. We measured ascorbate free radical, an index of free radical production, in the great cardiac vein effluent. Twenty-eight open-chest dogs underwent 20 min of coronary artery occlusion and 30 min of reperfusion. Ten dogs received no drug, 10 dogs received 750 mg bathocuproine, i.v., and eight dogs received 700 mg desferrioxamine, i.v. Both bathocuproine and desferrioxamine blunted the postreperfusion increase in ascorbate free radical generation: no drug, 36+/-8% increase; desferrioxamine, 13+/-5% increase; bathocuproine, 21+/-6% increase (p < 0.05 vs. baseline). Thus direct free radical measurements indicate that chelation of the transition metals iron and copper reduces free radical generation during reperfusion. PMID- 9733347 TI - Angiotensin-converting enzyme inhibition and salt in experimental myocardial infarction. AB - It is well known that angiotensin-converting enzyme inhibitors attenuate progressive ventricular enlargement or hypertrophy after myocardial infarction and that cardiac angiotensin-converting enzyme activity is increased in the rat model of myocardial infarction. In this study, to determine whether the beneficial effects of angiotensin-converting enzyme inhibition on cardiac hypertrophy after myocardial infarction are due to a reduction in ventricular afterload or to inhibition of cardiac angiotensin-converting enzyme, we used sodium loading during angiotensin-converting enzyme inhibition. The rat model of myocardial infarction was treated with a vehicle, 1% saline, as drinking fluid, perindopril (2 mg/kg/day), or 1% saline as drinking fluid plus perindopril (2 mg/kg/day) for 6 weeks. Perindopril reduced blood pressure, prevented cardiac hypertrophy, and inhibited cardiac angiotensin-converting enzyme. The effects of perindopril on blood pressure and cardiac hypertrophy were abolished by sodium loading, which did not alter the degree of cardiac angiotensin-converting enzyme inhibition. Thus the actions of perindopril on cardiac hypertrophy depend more on blood pressure reduction than on cardiac angiotensin-converting enzyme inhibition in the rat model of myocardial infarction. PMID- 9733348 TI - Evidence for constitutive release of nitric oxide in the venous circuit of pigs. AB - To determine whether the venous circuit constitutively produces nitric oxide (NO), we infused the NO synthase inhibitor, NG-nitro-L-arginine methyl ester (L NAME) into anesthetized and mechanically ventilated pigs and measured venous circuit parameters. We measured cardiac output (Q) by thermodilution and obtained arterial (Part), central venous pressure (in the inferior vena cava), right atrial (Pra), pulmonary artery (PAP), and pulmonary capillary wedge pressures. A balloon was transiently inflated in the right atrium to stop venous return and obtain mean circulatory filling pressure (MCFP). Venous compliance (Cv) was measured by volume boluses. Starling curves were obtained from changes in Q with changes in Pra from the boluses. Resistance to venous return (RVR) was calculated from (MCFP - Pra)/Q. After baseline measurements, we infused 25 mg/kg of L-NAME over 10 min in seven pigs and monitored them for a further 2 h. Three others served as time controls and showed no significant hemodynamic changes. L-NAME markedly decreased cardiac output from 3.8+/-0.86 to a low of 2.0+/-0.2 L/min, and increased blood pressure from 114+/-16 to 144+/-11 mm Hg and pulmonary artery pressure from 15+/-2 to 30+/-12 mm Hg (p < 0.05). MCFP increased from 9.1+/-1.2 to 11.4+/-2.4 mm Hg (p < 0.05); Cv did not change. Cardiac function curves were markedly depressed and flattened and remained depressed for 2 h. The increase in RVR of 167% from 1.8+/-0.6 mm Hg/L/min at baseline to 5.4+/-3.7 mm Hg/L/min (p < 0.05) was similar to the 188% increase in systemic vascular resistance. These data indicate that constitutive release of NO decreases baseline venous resistance and increases capacitance. There also appears to be a worsening of cardiac function when NOS is inhibited. PMID- 9733349 TI - ACE insertion/deletion genotype affects bradykinin metabolism. AB - A deletion allele of the angiotensin-converting enzyme (ACE) gene has been associated with increased serum ACE activity, enhanced conversion of angiotensin (Ang) I to Ang II, and cardiovascular morbidity. This study tested the hypothesis that the ACE deletion allele is also associated with enhanced degradation of bradykinin, a vasoprotective peptide. Metabolism of synthetic bradykinin was measured in sera obtained from subjects who were homozygous for either the ACE deletion (n=12) or insertion (n=8) allele and who had participated in an Ang I infusion protocol. ACE levels tended to be increased in subjects who were DD compared with those who were II [41.2 (95% CI, 27.9, 54.3) vs. 28.0 IU/L (20.0, 35.9); t=-1.6; p=0.1 18]. During Ang I infusion, plasma Ang II concentrations were increased in DD compared with II subjects (F =4.4; p=0.052). In contrast, the half-life of bradykinin was significantly decreased in sera obtained from ACE DD compared with II subjects [26.3 s (17.8, 34.6 s) vs. 42.1 s (24.4, 59.9 s); t= 2.4; p=0.029]. Moreover, there were significant inverse relations between the half-life of bradykinin and serum ACE activity (p < 0.001) and between the half life of bradykinin and the conversion of Ang I to Ang II (p=0.026). This study confirms that ACE genotype determines bradykinin degradation and suggests another mechanism whereby the ACE D allele could be associated with deleterious cardiovascular effects. PMID- 9733350 TI - Potentiation by isoniazid of relaxation induced by nitrovasodilators in rat aorta. AB - The influence of isoniazid (ISO) preincubation on the relaxant effects of a group of nitrovasodilators was examined in norepinephrine-contracted rat aortic rings with and without endothelium. ISO displaced to the left the dose-response curves to all nitrovasodilators and increased the negative logarithm of their median effective concentration (EC50) values. Potentiation was minimal with sodium nitrite and increased progressively with sodium nitroprusside, nitroglycerin (NTG), and isosorbide dinitrate. The phenomenon occurred in rings with and without endothelium and was not seen with the nitric oxide-releasing agent acetylcholine or with the nonspecific vasodilator hydralazine. These results confirm previous observations of potentiation of NTG vasorelaxation by ISO and extend them to other nitrovasodilators. Potentiation is attributed to the previously postulated inhibition by ISO of pyridoxal-requiring enzymes involved in the breakdown of homocysteine, leading to its intracellular accumulation. Increased availability of this sulfhydryl donor would lead in turn to enhanced bioactivation and vasorelaxant effects of nitrovasodilators. The different degrees of potentiation observed would be related to the sulfhydryl requirement of each compound for bioactivation. Alternatively, enhanced bioactivation of the nitrovasodilators could be due to induction by ISO of P-450 enzymes involved in this process. PMID- 9733351 TI - Ethanol selectively counteracts hypotension evoked by central I(1)-imidazoline but not alpha2-adrenergic receptor activation in spontaneously hypertensive rats. AB - Previous studies from our laboratory showed that ethanol counteracts hypotensive responses to clonidine in spontaneously hypertensive rats. This study investigated whether this effect of ethanol involves interaction with central alpha2-adrenoceptors or I(1)-imidazoline receptors or both. The effects of ethanol (0.5 or 1 g/kg, i.v.) or an equal volume of saline on hypotensive and bradycardic responses to clonidine (mixed alpha2-adrenoceptor/I(1)-imidazoline receptor agonist), rilmenidine (selective I(1)-imidazoline receptor agonist), or alpha-methylnorepinephrine (selective alpha2-adrenoceptor agonist) were studied in conscious spontaneously hypertensive rats. Intracisternal administration of clonidine (0.5 microg), rilmenidine (25 microg), or alpha-methylnorepinephrine (4 microg) elicited similar decreases in mean arterial pressure (MAP; 25-30 mm Hg) that lasted > or =60 min. Subsequent administration of ethanol (0.5 and 1 g/kg, i.v.) counteracted the hypotensive effect of clonidine in a dose-related manner. Ethanol (1 g/kg) increased the blood pressure to levels similar to baseline (preclonidine) levels, and blood pressure remained significantly (p < 0.05) higher compared with the corresponding values in saline-treated rats. Similarly, ethanol (0.5 and 1 g/kg, i.v.) dose-dependently counteracted the hypotensive effect of rilmenidine. The antagonizing effects of ethanol on hypotension evoked by clonidine and rilmenidine were comparable in terms of both magnitude and duration. In contrast, ethanol (0.5 or 1 g/kg) had no effect on hypotension evoked by alpha-methylnorepinephrine. Except for a brief increase in blood pressure by ethanol (1 g/kg) at 5 min, blood pressure values obtained in alpha methylnorepinephrine-treated rats receiving any of the two doses of ethanol were similar to postsaline values. Ethanol had no effect on bradycardic responses to any of the three hypotensive agents. Blood ethanol concentrations were similar regardless of the antihypertensive drug used. We concluded that the adverse hemodynamic effect of ethanol on centrally mediated hypotensive responses depends on the types of receptors involved in the elicitation of this response. That ethanol counteracts decreases in blood pressure evoked by clonidine and rilmenidine but not by alpha-methylnorepinephrine suggests an interaction between ethanol and central pathways involved in I(1)-imidazoline receptor-mediated hypotension. PMID- 9733352 TI - Ca2+ channel-blocking activity of propranolol and betaxolol in isolated bovine retinal microartery. AB - The relaxant action of the standard beta-blocker propranolol was compared with betaxolol, a beta-blocker with established vasorelaxant properties. Ring segments of bovine retinal microartery (n=36, theta=237 microm), which lacks adrenergic nerves and beta-adrenoceptors, were mounted in an organ bath for isometric force recording. l-, d-, dl-Propranolol and betaxolol were equally effective in relaxing tonic K+-induced contractions. The median effective dose (ED50) value was approximately 10(-5) M for both beta-blockers. The relaxation by both beta blockers was unaffected by endothelium removal. Like verapamil, both beta blockers induced smaller relaxation of tonic prostaglandin F2alpha (PGF2alpha) induced force, which depended less on Ca2+ influx than did K+-induced force: K+-, but not PGF2alpha-induced contractions were abolished in Ca2+-free medium. The minor betaxolol-induced relaxation of tonic PGF2alpha-induced force was blocked in Ca2+-free medium. With repeated exposures to PGF2alpha in Ca2+-free medium, initial phasic PGF2alpha-induced force declined less with every exposure than did subsequent tonic force. When the preparations were briefly equilibrated with K+- and Ca2+-rich solution before every exposure to PGF2alpha phasic force did not decline, indicating that phasic force primarily depended on Ca2+ released from intracellular stores. Both beta-blockers failed to relax phasic PGF2alpha-induced force. Thus propranolol and betaxolol are equipotent vasorelaxant drugs in retinal microartery, both probably acting via Ca2+ channel blockade. This activity (that shows no stereospecificity) thus appears to be a more general property of beta-blockers. Microarteries might be more sensitive to this activity than are conductance arteries. PMID- 9733353 TI - Pharmacokinetics and pharmacodynamics of sibrafiban (Ro 48-3657), an orally active IIb/IIIa antagonist, administered alone or in combination with heparin, aspirin, and recombinant tissue-type plasminogen activator in beagles. AB - This study characterized the pharmacokinetics (PK) and pharmacodynamics (PD) of sibrafiban (Ro 48-3657) in the presence of aspirin, heparin, and recombinant tissue-type plasminogen activator (rt-PA) in beagles. Sibrafiban is a double prodrug that undergoes bioconversion to the inactive prodrug Ro 48-3656 and to the active IIb/IIIa antagonist, Ro 44-3888, after oral administration. After oral sibrafiban, peak Ro 48-3656 plasma concentrations were observed earlier than Ro 44-3888 and were five- to sixfold higher than Ro 44-3888 peak concentrations. Administration of sibrafiban with heparin and aspirin or heparin and rt-PA did not alter sibrafiban PK. Ro 48-3656 and Ro 44-3888 PK and inhibition of platelet aggregation profiles in groups treated with sibrafiban and heparin/aspirin or sibrafiban and heparin/rt-PA were similar to those of the group receiving sibrafiban alone. Sibrafiban resulted in >80% inhibition of adenosine diphosphate (ADP)-mediated platelet aggregation and an approximate sixfold increase in bleeding time (BT) compared with baseline measurements. The BT increase was greater in the sibrafiban, heparin, and rt-PA-treated group, during rt-PA administration, compared with the group treated with sibrafiban alone. The recovery of platelet aggregation may be slower after administration of sibrafiban with heparin and rt-PA. Sibrafiban had no effect on rt-PA PK or heparin PD. PMID- 9733354 TI - Divergent effects of different antihypertensive drugs on endothelium-dependent vasodilation in the human forearm. AB - Several studies indicated an abnormal endothelium-dependent vasodilation (EDV) in hypertensive patients, but no study has systematically investigated the effects of different pharmacologic classes of antihypertensive drugs on EDV. This study aimed to evaluate the effects of three different antihypertensive regimens [angiotensin-converting enzyme (ACE) inhibition, calcium channel blockade, and beta-blockade] on EDV when given locally in the forearm at a constant blood pressure. The increase in forearm blood flow (FBF) during local intraarterial infusions of methacholine (MCh; inducing EDV) and sodium nitroprusside (SNP; inducing endothelium-independent vasodilation, EIDV) was measured in young, normotensive subjects by venous occlusion plethysmography, before and during concomitant local intraarterial infusion of any of the antihypertensive drugs. Without changing baseline FBF, enalaprilat (n=6, 2.4 mg/h) potentiated the increase in FBF induced by MCh [from 22.6+/-2.3 (SD) to 25.4+/-2.3 ml/min/100 ml tissue at 4 microg/min; p < 0.05], but the response to SNP was unchanged. Local intraarterial verapamil infusion (n=6), at a dose individually titrated to keep baseline FBF unchanged, did not alter the response to MCh infusion, whereas the response to SNP was potentiated. A higher dose of verapamil (n=6), which increased baseline FBF, increased both EDV and EIDV significantly in parallel (p < 0.05). The local propranolol infusion (n=6, 1.2 mg/h) attenuated the FBF response to MCh significantly (from 28.9+/-5.7 to 21.5+/-3.2 ml/min/100 ml tissue at 4 microg/min; p < 0.05), whereas both baseline FBF and the response to SNP were unchanged. In conclusion, this investigation showed that commonly used antihypertensive drugs affect endothelial vasodilator function in a different ways. ACE inhibition enhanced EDV, whereas a nonselective beta-blocker attenuated EDV. The calcium channel blocker, verapamil, improved both EDV and EIDV, probably by a direct effect on the vascular smooth-muscle cells. PMID- 9733355 TI - Local cholinergic suppression of pacemaker activity in the rabbit sinoatrial node. AB - The effects of transmural vagal stimulation and acetylcholine (ACh) superfusion on primary and latent pacemaker cells of the rabbit sinoatrial node were studied by using microelectrodes. Both ACh and vagal stimulation lengthened atrial cycle length by 40-60% as compared with control. In the cells from the primary pacemaker area, both ACh superfusion and vagal stimulation suppressed action potential (AP) amplitude and then induced inexcitability. In contrast, cells from subsidiary pacemaker area as well as atrium remained excitable. These effects were completely reversible and also were abolished by atropine, 10(-7) M. Cholinergically induced suppression of AP amplitude is predictable based on the maximal rate of AP upstroke (dV/dt). The probability of amplitude suppression was the highest among pacemaker cells (dV/dt, <3 V/s), in which ACh suppressed amplitude in 27 (93%) of 29 cells, and vagal stimulation did so in 38 (81%) of 47 cells. With increasing upstroke velocity, the probability of amplitude suppression decreased. Inexcitability did not occur in cells whose dV/dt was >15 V/s. The suppression of AP amplitude by ACh occurred in a concentration-dependent manner: the concentration inducing suppression of amplitude in 50% of pacemaker cells was approximately 10 microM. These results indicate that cholinergic effects on typical pacemaker and subsidiary pacemaker cells are different: whereas subsidiary pacemaker cells remain excitable, typical pacemaker cells become quiescent. We hypothesize that quiescent cells create quiescent regions in the center of the sinoatrial node that might functionally be an obstacle for reentrant tachycardias. PMID- 9733356 TI - Drug-related torsades de pointes in the isolated rabbit heart: comparison of clofilium, d,l-sotalol, and erythromycin. AB - Torsades de pointes is a potentially life-threatening form of polymorphic ventricular tachyarrhythmia typically seen in the presence of repolarization prolonging agents. We investigated this particular form of tachyarrhythmia in the isolated, perfused rabbit heart. The experimental model was designed to reproduce conditions that are clinically known to be associated with an increased propensity to the development of torsades de pointes. The class III agent clofilium (1 microM) and d,l-sotalol (10 microM), as well as the antibiotic erythromycin (30-150 microM) were infused in the presence of either normal (5.88 mM) or low (1.5 mM) potassium concentration in sinus-driven or atrioventricular (AV)-blocked hearts. Ventricular tachyarrhythmias spontaneously emerged in the clofilium-, d,l-sotalol-, and erythromycin-treated AV-blocked hearts. The episodes showed typical features of torsades de pointes found in humans. They developed within 4-12 min after the onset of infusion, were normally nonsustained, and only rarely degenerated into ventricular fibrillation. Electrical stimulation at cycle lengths <600 ms and perfusion with MgSO4 suppressed arrhythmic activity. In the d,l-sotalol- and erythromycin-treated hearts, torsades de pointes occurred only in the presence of hypokalemia and bradycardia, whereas, in the presence of clofilium, bradycardia alone caused torsades de pointes. Monophasic action-potential recordings demonstrated early afterdepolarizations in endocardial and epicardial recordings. Thus the isolated AV-blocked rabbit heart represents a model for studying drug-related torsades de pointes and its mechanism. PMID- 9733357 TI - Effects of sumatriptan on coronary flow and left ventricular function in the isolated perfused guinea pig heart. AB - The effects of the 5-HT1B/D receptor agonist, sumatriptan, on coronary flow (CF) and left ventricular function in the isolated perfused guinea pig heart were investigated in the presence and absence of coronary endothelial dysfunction induced by nitric oxide (NO) synthase inhibition with Nomega-nitro-L-arginine methyl ester (L-NAME; 10 microM). Hearts were perfused under constant pressure (80 cm H2O) with oxygenated (95% O2/5% CO2) Krebs bicarbonate buffer (pH 7.4) and were driven at 4 Hz. In the absence of L-NAME (n=37), sumatriptan (0.1-32 microM) failed statistically significantly to affect left ventricular developed pressure (LVDP; maximal change, -8.1+/-1.8%; NS vs. vehicle), left ventricular end diastolic pressure (LVEDP; +10.4+/-9.8%, NS), or CF (-12.2+/-1.4%; NS compared with vehicle). L-NAME per se significantly reduced coronary flow (CF; -26.3+/ 2.9%; p < 0.001), thereby increasing coronary vascular tone, and decreased LVDP ( 17.1+/-1.8%; p < 0.01). In hearts perfused with L-NAME (10 microM; n=61), sumatriptan (0.1-32 microM) still failed significantly to affect CF (maximal change, 0.2+/-5.7%, NS) but concentration-dependently increased LVEDP [maximal increase, 89.0+/-30.3%; p < 0.05; geometric mean EC50 3.6 (2.9-5.7) microM], which was not prevented by the 5-HT1B/D receptor antagonist, GR 127935 (0.1 microM; maximal increase, 51.8+/-11.1%; n=48, NS compared with sumatriptan alone). In conclusion, sumatriptan failed significantly to affect CF even in the presence of endothelial dysfunction. LV function similarly remained unaffected in normal hearts, but sumatriptan produced diastolic contracture in the presence of coronary endothelial dysfunction by a mechanism apparently not involving 5-HT1B/D receptors. Collectively the data indicate that 5-HT1B/D receptor expression or effector coupling or both are absent or low in the guinea pig heart, because no detectable functional responses were observed. PMID- 9733358 TI - Comparative effects of carvedilol and metoprolol on cardiac ischemia-reperfusion injury. AB - The effects of carvedilol, a multiple-action neurohormonal antagonist, and metoprolol, a highly selective beta1 antagonist, were compared on postischemic contractile recovery and contracture. Isolated rabbit hearts were aerobically perfused for 45 min and subjected to zero-flow normothermic ischemia for 30 or 60 min followed by reperfusion for 30 min. Carvedilol and metoprolol were added to the perfusion solution 10 min before inducing ischemia and were maintained in the perfusate throughout reperfusion. Left ventricular developed pressure (LVDP) and left ventricular end-diastolic pressure (LVEDP) were assessed with an intraventricular balloon. Because the volume of the balloon was held constant, an increase in LVEDP reflected an increase in diastolic chamber stiffness or "contracture." After 30 min of ischemia, the carvedilol-treated hearts exhibited a significantly better cardiac function than did control or metoprolol-treated hearts. At the end of reperfusion, the control group LVDP recovered to 21.4+/ 9.9% of the preischemic value. With 0.03, 0.1, and 0.3 microM metoprolol, LVDP recovered to 33.2+/-13.6%, 41.7+/-13.0%, and 48.8+/-13.3% of initial developed pressure, respectively. In the carvedilol group, a greater recovery of LVDP was obtained at 0.03, 0.1, and 0.3 microM: 64.0+/-2.5%, 60.4+/-6.3%, and 68.0+/-2.0% of preischemic values, respectively (p < 0.05 vs. controls). Within the first 5 min of reperfusion, LVEDP increased to 70.3+/-2.7 mm Hg in control hearts, indicating a pronounced contracture, whereas metoprolol reduced LVEDP when given at high concentration, 0.3 microM (41.9+/-10.7 mm Hg). Carvedilol, even at the lowest concentration, 0.03 microM, almost completely inhibited the postischemic contracture (16.5+/-4.0 mm Hg; p < 0.05 vs. control and metoprolol). The cardioprotection provided by carvedilol also is observed in hearts subjected to more severe ischemic periods. After 60 min of ischemia, control hearts failed to restore LVDP function; in the metoprolol group, ventricular function recovered to only 4.6+/-3.1%, whereas carvedilol-treated hearts exhibited 23.6+/-1.9% of preischemic values at the end of reperfusion. In addition, carvedilol induced a reduction in ischemic contracture: control, 36.7+/-3 mm Hg; metoprolol, 38.7+/ 3.7 mm Hg; and carvedilol, 15.7+/-8.4 mm Hg at 50 min of ischemia. Similarly, carvedilol reduced contracture during the reperfusion compared with metoprolol and control groups (83.2+/-3.4 mm Hg, 106.9+/-3.3 mm Hg, and 107.6+/-4.1 mm Hg, respectively). These data clearly demonstrate that carvedilol was markedly more effective than metoprolol to protect systolic function after ischemia and to reduce postischemic contracture. PMID- 9733359 TI - Bradykinin B2-receptor-mediated positive chronotropic effect of bradykinin in isolated rat atria. AB - The positive chronotropic effect of bradykinin was investigated in isolated spontaneously beating atria of the rat. Cumulative additions of bradykinin (0.3 100 nM) caused a concentration-dependent increase in the beating rate of the atria by maximally 35+/-4 beats/min, approximately 25% of the 1 microM isoprenaline-induced maximal responses. In contrast, the active metabolite of bradykinin and selective bradykinin B1-receptor agonist, Des-Arg9-bradykinin, did not influence the spontaneous frequency of beating. Propranolol (1 microM) combined with prazosin (1 microM) did not affect the positive chronotropic effect of bradykinin. A selective bradykinin B2-receptor antagonist, Hoe 140, concentration-dependently shifted the response curves for bradykinin to the right, whereas the bradykinin B1-receptor antagonist, Lys-[Leu8]Des-Arg9 bradykinin had no effect. The tachycardic responses to bradykinin were potentiated by ramipril, an angiotensin-converting enzyme/kininase II inhibitor, but not affected by Nomega-nitro-L-arginine methyl ester hydrochloride, a nitric oxide synthesis inhibitor. Indomethacin and meclofenamate, two cyclooxygenase inhibitors, abolished the bradykinin-induced chronotropic effect. These results indicate that exogenous bradykinin induces a positive chronotropic effect that occurs independent of adrenoceptors. The bradykinin-induced chronotropic effect is mediated by bradykinin B2 receptors, whereas B1 receptors do not play a role in mediating this effect. Prostaglandins but not nitric oxide appear to be involved in bradykinin-induced positive chronotropic effect. PMID- 9733360 TI - 17-Beta estradiol regulation of myocardial glutathione and its role in protection against myocardial stunning in dogs. AB - We studied the effect of 2-week treatment with estradiol 17beta on myocardial glutathione concentration in dogs and isolated perfused rat heart subjected to brief coronary ischemia and reperfusion. Estradiol protected against ischemia/reperfusion-induced myocardial systolic shortening and malonylaldehyde production and increased myocardial glutathione concentration and glucose-6 phosphate dehydrogenase enzyme activity. Reduction of myocardial glutathione with buthionine sulfoximine to levels seen in the absence of estrogen reversed the protective effect of estradiol against myocardial dysfunction and lipid peroxidation associated with ischemia/reperfusion. These results suggest that the antioxidant effect of estradiol in ischemia/reperfusion may be mediated by regulation of myocardial glutathione metabolism. PMID- 9733361 TI - Effects of balanced vasodilator, flosequinan, on aortic impedance in failing heart. AB - An arteriovenous vasodilator, flosequinan, has been shown to be effective for the treatment of acute heart failure. However, little is known as to its effect on aortic impedance, which is known to be a proper and precise expression of left ventricular (LV) afterload. To evaluate the acute cardiovascular effect of flosequinan in failing heart, we administered flosequinan intravenously to seven dogs with cardiac failure produced by an infusion of carbon powder (20-50 microm in diameter) into left main trunks of coronary artery. The LV-pump function was severely impaired after intracoronary injection of carbon powder, as evidenced by the findings that cardiac output, circumferential shortening velocity (mean Vcf), and peak +dP/dt of LV pressure were all decreased, associated with a significant increase in LV end-diastolic pressure. Flosequinan (0.9 mg/kg, i.v.) increased cardiac output by 28%, mean Vcf by 44%, and peak +dP/dt by 24%, whereas it decreased total systemic resistance by 32%, time constant of LV pressure decay by 22%, and LV end-diastolic pressure by 18%. Moreover, flosequinan substantially decreased the pulsatile components of LV afterload (i.e., characteristic impedance by 11% and arterial wave reflection coefficient by 45%). Thus flosequinan exerted not only positive inotropic but also positive lusitropic effects, in association with a significant reduction of both pulsatile and steady components of LV afterload, contributing to an improvement of LV-pump function in acute cardiac failure. PMID- 9733362 TI - Actions of 8 epi prostaglandin F2alpha on isolated rat aorta. AB - 8-epi prostaglandin F2alpha(8-epi PGF2alpha) contracted rat thoracic aorta rings in a concentration-dependent manner in the presence or absence of functional endothelium [median effective concentration (EC50) values, 455+/-52 and 268+/-34 nM, respectively; Student's t test; p=0.006]. U46619 was a more potent agonist with or without functional endothelium (EC50 values, 6.8+/-1.6 and 4.5+/-1.0 nM, respectively). SQ29548 [a thromboxane (TP)-receptor antagonist] inhibited contractions to both 8-epi PGF2alpha and U46619 in a competitive manner, with mean pA2 values of 8.3 and 7.9, respectively. 8-Epi PGF2alpha had a further contractile effect in vessels that had been contracted with noradrenaline and had been shown to possess a functional endothelium. Inhibition of thromboxane synthesis with OKY-046 or blockade of endothelin receptors with bosentan had no effect on responses to 8-epi PGF2alpha or U46619. Preincubation with 8-epi PGF2alpha or noradrenaline shifted the concentration-response curves to U46619 upward at low concentrations of U46619 with no significant change in EC50 values or maximal responses. Reduction of TP-receptor number in rat aorta with dithiothreitol caused a concentration-dependent inhibition of responses to both U46619 and 8-epi PGF2alpha, with no effect on maximal responses and or on the responses to U46619 after the preincubation with 8-epi PGF2alpha. These results indicate that 8-epi PGF2alpha is a potent vasoconstrictor in the rat aorta and are suggestive of an action of 8-epi PGF2alpha at the TP receptor. PMID- 9733364 TI - Pharmacokinetics and pharmacodynamics of SM-20302, a GPIIb/IIIa receptor antagonist, in anesthetized dogs. AB - We examined the pharmacokinetic and pharmacodynamic properties of SM-20302, a GPIIb/IIIa receptor antagonist, in anesthetized dogs. SM-20302 was administered intravenously in doses of 30 (n=2), 100 (n=4), 300 (n=4), and 1,000 microg/kg (n=4). The half-life of the initial phase was 4 min, and that of the terminal phase was 162-209 min. SM-20302 produced a dose-dependent increase in the initial plasma concentration and the area under concentration-time curve but did not alter the volume of distribution, mean residence time, or plasma clearance. Plasma clearance for SM-20302 ranged from 6.58 to 9.73 ml/min/kg. All doses of SM 20302 inhibited (> or =90%) the ex vivo platelet aggregation induced by adenosine diphosphate (ADP) or arachidonic acid (AA) in citrated platelet-rich plasma (cPRP). In heparinized PRP (hPRP), a dose-dependent (44-89%) inhibition was observed. By using a sigmoid Emax model, the in vivo median inhibitory concentration (IC50) for SM-20302 was estimated to be 14-19 ng/ml in cPRP and 79 89 ng/ml in hPRP. To validate the calculated parameters, an infusion regimen was designed for the prevention of coronary artery thrombosis. Infusion of SM-20302 produced 64-67% inhibition of platelets in hPRP and maintained vessel patency despite vessel wall injury. The results suggest that SM-20302 exhibits linear pharmacokinetics and that its ability to inhibit platelet aggregation in hPRP may correlate more accurately with its in vivo antithrombotic efficacy. PMID- 9733363 TI - Effect of cilnidipine, a novel dihydropyridine Ca2+ channel blocker, on adrenal catecholamine secretion in anesthetized dogs. AB - We investigated the effect of cilnidipine, a novel dihydropyridine Ca2+ channel blocker possessing blocking actions on N-type and L-type voltage-dependent Ca2+ channels (VDCCs), in comparison with the L-type VDCC blocker nifedipine, on adrenal catecholamine secretion in response to splanchnic nerve stimulation (SNS), acetylcholine (ACh), the nicotinic receptor stimulant 1,1-dimethyl-4 phenyl-piperazinium (DMPP), and muscarine in anesthetized dogs. Ca2+ channel blockers and cholinergic agonists were infused and injected, respectively, into the adrenal gland through the phrenicoabdominal artery. Cilnidipine (0.3-3 microg/min) inhibited increases in both epinephrine (EPI) and norepinephrine (NE) output induced by SNS (2 Hz), ACh (1.5 microg), and DMPP (0.2 microg). However, cilnidipine inhibited increase in NE output induced by muscarine (1 microg) without affecting increase in EPI output. Nifedipine (0.3-3 microg/min) inhibited the ACh- and DMPP-induced increases in EPI and NE output without affecting the SNS- and muscarine-induced increases in EPI and NE output. From these results, it seems likely that the inhibition by cilnidipine of the SNS-induced EPI and NE secretion and of the muscarine-induced NE secretion is related to its blocking action on N-type VDCCs. PMID- 9733366 TI - Aggravation of myocardial infarction in the porcine heart by capsaicin-induced depletion of calcitonin gene-related peptide (CGRP). AB - The potent vasodilator calcitonin gene-related peptide (CGRP) is stored in a population of C-fiber afferents that are sensitive to capsaicin. CGRP has been suggested to have a beneficial effect in myocardial ischemia. In this study we used capsaicin pretreatment to deplete cardiac C-fiber peptide stores and tried to evaluate the role of endogenous CGRP in myocardial ischemia. Six pigs were pretreated with capsaicin (50 mg/kg). Forty-eight hours later, they were subjected to 40min occlusion of the left anterior descending coronary artery. After 4 h of reperfusion, the heart was excised, and the extent of myocardial infarction was measured by using triphenyl tetrazolium chloride. Content of CGRP in the ischemic and the nonischemic myocardium was measured by radioimmunoassay. Capsaicin-treated pigs had more extensive myocardial infarction (56+/-6% vs. 26+/ 8% of the area at risk; p=0.013) and a lower myocardial content of CGRP (14+/-6 vs. 32+/-5 pmol/g; p=0.039) compared with six untreated control pigs. Furthermore, capsaicin-treated pigs had significantly increased mean arterial blood pressure compared with controls. This study indicates that peptides released from cardiac C fibers have a beneficial effect in myocardial ischemia and reperfusion. In view of its potent effects in cardiovascular regulation, CGRP is a possible candidate for the mediation of the observed cardioprotective effect. PMID- 9733365 TI - Effects of moxonidine on stress-induced peak blood pressure and renal function: a randomized, double-blind, placebo-controlled crossover study. AB - Moxonidine is an imidazoline I1-receptor agonist that centrally acts by reducing the sympathetic tone. Furthermore, proximal tubular I1-receptors have been isolated in human kidneys, but their natriuretic effects have never been demonstrated. Because stress tests elicited a sympathetically mediated increase in blood pressure and in sodium reabsorption, the aim of this study was to assess the effects of moxonidine (0.4 mg/day; 1 month) on stress-induced cardiovascular response and renal sodium handling in hypertensives, in a double-blind, crossover, placebo-controlled study. The stress test used is an efficient and reproducible computerized version of Stroop's stress test. During the experimental sessions, both rest and stress renal functional parameters were determined: glomerular filtration rate (inulin clearance), renal plasma flow (para-aminohippurate clearance), filtration fraction, sodium excretion, and segmental sodium tubular reabsorption (lithium clearance). During the placebo phase, stress induced a significant increase in systolic blood pressure (deltaSBP; 15.8+/-10.7 mm Hg) and diastolic blood pressure (deltaDBP; 8.2+/-6.1 mm Hg). During stress, glomerular filtration rate tended to decrease, whereas renal plasma flow significantly decreased, resulting in a significant increase in filtration fraction. Despite the increase in BP, stress induced a decrease in sodium excretion that was mainly due to a nonsignificant increase in sodium reabsorption in the proximal parts of the tubules. Moxonidine significantly reduced rest and stress BP, but the stress cardiovascular reactivity was not altered. At rest, renal function was well preserved by the treatment. Stress induced modifications in renal function and sodium handling were not altered by the treatment. In conclusion, moxonidine reduced rest and stress-induced peak BP and preserved basal renal function. The study failed to demonstrate any effect of moxonidine either on basal renal sodium handling or on stress-induced increase in sodium reabsorption. PMID- 9733367 TI - Aggravation of left ventricular remodeling by a novel specific endothelin ET(A) antagonist EMD94246 in rats with experimental myocardial infarction. AB - An endothelin (ET(A)) antagonist reduced mortality and an ET(A) + ET(B) antagonist prevented left ventricular dilatation in rats with large myocardial infarction. This study tested the hypothesis that long-term blockade of the ET(A) receptor would have beneficial effects on left ventricular function and remodeling. Three hours after coronary artery ligation or sham operation in rats, EMD94246 (100 mg/kg/day, n=62) or placebo (n=62) was given by gavage. Eight weeks later, left ventricular hemodynamic measurements were performed and left ventricular volume determined with a double-lumen catheter after KCl-induced cardiac arrest. EMD94246 treatment had no effects on mortality or hemodynamic parameters. In rats with large infarcts, EMD94246 significantly increased left ventricular volume (2.5+/-0.1 vs. 2.2+/-0.1 ml/kg; p < 0.05). The nonpeptide ET(A)-selective antagonist EMD94246 promoted chronic left ventricular dilatation in rats with large myocardial infarction. PMID- 9733368 TI - Parasitic infections of the heart. PMID- 9733369 TI - Multi-resistant pneumococci in children in day-care centres. PMID- 9733370 TI - Drug-resistant Streptococcus pneumoniae in day-care centres in Stockholm County. AB - Between January 1994 and July 1995, 40 pre-school children were found to have drug-resistant Streptococcus pneumoniae(DRSP), i.e. reduced sensitivity to penicillin (minimum inhibitory concentration, MIC, > or =0.1) and resistance to at least two other antibiotic drugs. Twenty-five of the children were index cases with symptoms of respiratory disease, and 15 children were carriers discovered in contact-tracing in connection with an index case. Children attending the same group in the day-care centre as an index child were routinely screened. Thirteen of the index children were attending day-care centres. In 11 of these day-care centres, contact-tracing and nasopharyngeal swabs from 424 children and 128 day care personnel identified an additional 13 asymptomatic children who were carriers of DRSP. In all but one case, the same serotype as the index case was discovered. No day-care personnel were carriers of the DRSP strain. Sixteen (64%) of the 25 children with symptoms caused by DRSP were 1 year old or younger, whereas eight (61%) of the 13 children who were carriers of DRSP were 3-4 years old. In conclusion, when a child attending a day-care centre is discovered to have respiratory disease caused by DRSP, there is a great probability that additional children will be identified in the group with the same DRSP strain. PMID- 9733371 TI - A retrospective study of treatment of cerebral toxoplasmosis in AIDS patients with trimethoprim-sulphamethoxazole. AB - AIMS OF THE STUDY: a retrospective study was designed to evaluate efficacy and tolerance of trimethoprim-sulphamethoxazole (TMP-SMZ) in AIDS patients with cerebral toxoplasmosis (TE). PATIENTS AND METHODS: we reviewed 471 patients with AIDS, and we analysed 71 AIDS patients with TE, who received intravenous therapy with TMP-SMZ (TMP: 10 mg/kg/day, SMZ: 50 mg/kg/day) for 4 weeks. RESULTS: 35 patients (49.2%) had a complete regression of clinical signs, and a complete resolution of radiological lesions was noted in 41 patients (57.7%). Improvement of clinical signs and radiological lesions were observed in 27 patients (38%), and in nine patients (12.6%), respectively. In contrast, nine patients (12.6%) did not show any clinical change, or worsened. Twenty-two patients (30.9%) suffered from adverse cutaneous reactions, whereas many patients had haematological toxicity. CONCLUSIONS: TMP-SMZ seems to be an efficient therapy for TE in AIDS patients, although further prospective, randomized therapeutic trials are required to confirm these results. PMID- 9733372 TI - Natural and 'in vitro' selected antigenic variants of influenza A virus (H2N2). AB - We provide data on the prevalence of SRH antibody to influenza A/Singapore/1/57 (H2N2). Approximately 10.3% of sera had antibody to the influenza A (H2N2) subtype virus in comparison to the 36.9% of positive sera to a representative influenza A (H3N2) and 31.5% to influenza A (H1N1) viruses. The percentage of subjects with antibody constantly decreased from the older to the younger age groups. Persons born after 1968 were essentially seronegative, whereas subjects born before 1900, and in the decade 1950-1959, showed the highest antibody levels to influenza A (H2N2) viruses. These age groups also appeared to have 'protective' levels of anti-HA antibody to influenza A (H2N2) virus. An antigenic variant of A/Singapore/1/57 virus was selected in the laboratory using a monoclonal antibody to HA. Serological comparison of the new in vitro variant with the parental virus and two naturally occurring viruses, namely A/England/12/64 and Tokyo/3/67, showed that certain human sera were able to distinguish the variant, indicating a restricted antibody repertoire in these adult and children's sera, providing an explanation of how such variants could actually arise in nature. PMID- 9733373 TI - Primary cutaneous aspergillosis: our experience in 10 years. AB - We describe nine patients with primary cutaneous aspergillosis who were diagnosed during the 10-year period between 1987 and 1996. All except one were adults. Seven of those nine cases had an immunocompromised state. Among the immunocompromised patients, six had burn wounds and one was a renal transplant recipient. Aspergillus flavus was isolated from seven patients and A. fumigatus from one, and in one other case the species could not be determined. Three patients who had total burn surface area of more than 70% died. The other patients responded well to extensive surgical debridement with or without institution of oral itraconazole. PMID- 9733374 TI - The incidence of, and factors leading to, parvovirus B19-related hydrops fetalis following maternal infection; report of 10 cases and meta-analysis. AB - OBJECTIVES: to clarify the approximation of the frequency of B19-related nonimmune hydrops fetalis (NIHF), and to know the critical period during which maternal infection led to NIHF. METHODS: we investigated the characteristics of 10 cases of antenatal B19 infection diagnosed over the past 10 years in Miyagi prefecture, Japan, and performed a meta-analysis of these cases and those previously reported in the literature. RESULTS: NIHF caused by intrauterine B19 infection was diagnosed between 11 and 23 weeks of gestation in 10 women over the past 10 years in Miyagi prefecture, Japan. The source of infection was the mother's older child in six out of 10 cases, and children at a kindergarten where the mothers worked in two cases. The interval between the onset of infection and the diagnosis of NIHF ranged from 2 to 6 weeks. B19 infection was responsible for 10 (15.2%) in 66 cases of aetiology unknown NIHF in this study, and for 57 (19.1%) of 299 cases of non-malformed or aetiology-unknown NIHF by meta-analysis of the literature. Meta-analysis of the 165 reported cases of antenatal B19 infection, including the 10 cases described above, showed that there was a 10.2% excess risk of fetal death in women infected with B19 during pregnancy and a 12.40% excess risk in women infected during the first 20 weeks of pregnancy. Transplacental transmission was confirmed in 69 (24.1%) of 286 cases. The mean gestational age at diagnosis of NIHF was 22.8 +/- 5.1 weeks. The mean interval between the onset of maternal infection and diagnosis of NIHF was 6.2 +/- 3.7 weeks. CONCLUSIONS: these approximations will be useful for counselling and management for pregnant women. The critical period during which maternal infection led to NIHF correlated with the hepatic period of hematopoietic activity. These findings suggest that parvovirus B19 may have an affinity for erythroid lineage cells at the hepatic stage of hematopoiesis, which may strongly influence the clinical features of feto-maternal B19 infection. PMID- 9733375 TI - A retrospective study on the efficacy and safety of amphotericin B in a lipid emulsion for the treatment of cryptococcal meningitis in AIDS patients. AB - To evaluate the efficacy and safety of Amphotericin B dissolved in dextrose (Amb) or in a lipid emulsion (Intralipid, Amb-IL) in AIDS patients with cryptococcal meningitis, we conducted a retrospective study in 30 AIDS patients with cryptococcal meningitis. A clinical complete resolution was obtained in 11 patients (55%) treated with Amb, and in six patients (60%) treated with Amb-IL. Intralipid did not decrease the infusion-related adverse effects, in particular nephrotoxicity and anaemia. Our results indicate that Amb-IL formulation is useful in the treatment of cryptococcal meningitis in AIDS patients, but it does not reduce the infusion-related adverse events. PMID- 9733376 TI - Is new immigrant screening for tuberculosis still worthwhile? AB - BACKGROUND: new immigrant screening in the 1980s showed that the official Port of Arrival (POA) system performed poorly, that there was a significant yield in terms of clinical tuberculosis, and that preventive measures (chemoprophylaxis and BCG) were appropriate in a substantial minority Data for the 1990s was sought for comparison. METHODS: prospective data on new immigrant screening for 1990 1994 inclusive in the Blackburn, Hyndburn and Ribble Valley local government areas were analysed, and compared with previous 1983-1988 data. RESULTS: of the 2242 new immigrants screened, 1333 were from Pakistan, 604 from India and 305 from the rest of the world. A total of 898 (40%) were found via the POA system, but 1344 (60%) were only identified by local links with the Family Health Services Authority (FHSA). Ten cases of active tuberculosis were found (0.45%), chemoprophylaxis was given to 19/465 (4.1%) of children aged 0-15 years, and BCG vaccination to 530/1705 (31%) of those aged under 30 years. CONCLUSIONS: between 1990-1994 the official POA system continued to perform poorly. The yield of new tuberculosis cases detected was lower than in the 1980s. Chemoprophylaxis at 4% and BCG vaccination at 31% showed that preventive health measures were appropriate for over one-third of new immigrants aged under 30. PMID- 9733377 TI - Regional microbiology of the cystic fibrosis lung: a post-mortem study in adults. AB - Although the majority of patients with cystic fibrosis (CF) become chronically colonized with Pseudomonas aeruginosa, the mode of acquisition of infection remains unclear. Epidemiological studies using genotyping techniques suggest that person-to-person transmission of this organism may occur. All these studies have utilized sputum or throat swab samples. We have studied the regional microbiological flora of the lungs of five CF patients at post-mortem and genotyped P. aeruginosa strains found therein and in the major airway. We have shown that although in most cases major airway secretions accurately reflect the peripheral lung flora, in cases of multiple strain carriage, major airway cultures may not reflect all strains present in the periphery of the lung. This finding has implications for the interpretation of epidemiological studies that use genotyping of strains from sputum and throat swab samples to assess possible routes of transmission. PMID- 9733378 TI - Incidence of gonorrhea in Belgrade, 1988-1994. AB - The incidence of gonorrhea in Belgrade decreased 55.5% from 1988 to 1994, but a decreasing trend of gonorrhea incidence actually started many years before. In all age groups gonorrhea incidence was higher in men than women, and the male/female ratio increased with age. The incidence was highest in men and women 20-29 years old. In both sexes the lowest incidence was below 15 years of age and in persons 50 or more years of age. In men, service and industrial workers were the most frequently affected by gonorrhea. In women the disease was commonest among unemployed persons and among workers of the service and industrial sectors. In both sexes gonorrhea infection was commonest in populations who had never married, and in the divorced population. PMID- 9733379 TI - Chronic diarrhoea among HIV-infected adult patients in Nairobi, Kenya. AB - OBJECTIVES: Chronic diarrhoea and wasting are well recognized features of AIDS in Africa. However, because of resource constraints few comprehensive aetiological studies have been conducted in sub-Saharan Africa which have included a broad range of microbiological investigations. We undertook a prospective cross sectional study of adult patients admitted to a government hospital in Nairobi, Kenya, to determine possible bacterial, mycobacterial, parasitic and viral causes of diarrhoea; to consider which may be treatable; and to relate microbiological findings to clinical outcome. METHODS: Stool specimens from 75 consecutive HIV seropositive patients with chronic diarrhoea admitted to a Nairobi hospital were subjected to microbiological investigation and results were compared with clinical findings and outcome. Stool samples were cultured for bacteria and mycobacteria and underwent light and electron microscopy; lawns of Escherichia coli were probed for pathogenic types and aliquots were tested for the presence of Clostridium difficile cytotoxin. Blood cultures for mycobacteria and other bacterial pathogens were performed as clinically indicated. RESULTS: Thirty-nine (52%) patients yielded putative pathogens, the most common being Cryptosporidium sp. (17%), Salmonella typhimurium (13%), and Mycobacterium tuberculosis (13%). Of 41 patients investigated for pathogenic Escherichia coli, enteroaggregative E. coli and diffusely adherent E. coli were each found in four patients. Thirty-one (41%) patients died. Detection of cryptosporidium cysts was the single most significant predictor of death (X2 = 5.2, P<0.05). Many patients did not improve (21; 28%) or self-discharged whilst still sick (5; 7%) but five (7%) were diagnosed ante mortem with tuberculosis and treated and a further 13 (17%) showed improvement by time of discharge. CONCLUSIONS: HIV-infected patients with chronic diarrhoea in Nairobi have a poor outcome overall, and even with extensive investigation a putative pathogen was identified in only just over half the patients. The most important step is to exclude tuberculosis; and the most useful investigation appears to be Ziehl-Neelsen staining. Other potentially treatable gram-negative bacterial pathogens, S. typhimurium, Shigella sp. and adherent E. coli were, however, common but require culture facilities which are not widely accessible for definitive identification. Further studies focussing on simple ways to identify sub-groups of patients with treatable infections are warranted. PMID- 9733380 TI - Hospital admissions in children due to pneumococcal pneumonia in England. AB - Hospital records of 116 children under 5 years of age discharged from 11 hospitals in three regions in England with a diagnosis of lobar (pneumococcal) pneumonia were reviewed to estimate the proportion likely to be attributable to infection with Streptococcus pneumoniae. Of these, 100 (86%) had lobar/focal changes on chest X-ray consistent with pneumococcal infection, although only one (1%) had pneumococcus isolated from blood. However, a further 89 (89%) with a lobar/focal picture were considered to be likely or possibly due to pneumococcal infection on the basis of the white cell count, level of C-reactive protein, isolation of the S. pneumoniae from either sputum or nasopharingeal aspirate and failure to identify another responsible pathogen. Of 135 cases with a discharge diagnosis of bronchopneumonia or pneumonia (organism unspecified), two (1%) had S. pneumoniae isolated from blood and a further 95 (70%) had clinical or laboratory features consistent with pneumococcal infection or S. pneumoniae isolated from either sputum or nasopharyngeal aspirate. With the imminent availability of conjugate pneumococcal vaccines, there is a need for improved diagnostic methods for identifying the pathogens responsible for community acquired pneumonia in young children. PMID- 9733381 TI - Paediatric neurobrucellosis: case report and literature review. AB - Neurological complications are rare in childhood brucellosis: there are only 33 reported cases. In children, neurobrucellosis is usually of acute presentation involving the central nervous system. We report our experience with an 8-year-old boy with brucella meningitis who demonstrated a Jarisch-Herxheimer-like reaction, i.e. initial clinical deterioration following the commencement of antibrucella treatment, associated with increased pleocytosis and shift from lymphocytic to polymorphic predominance and an already increased CSF lactate. These CSF findings have not been previously described. The patient recovered completely after 3 months' therapy consisting of rifampicin, doxycycline and gentamicin. Paediatric neurobrucellosis therapy should be a combination of three antibrucella antibiotic that include an aminoglycoside; for a period of 8-12 weeks, steroids may be added to treat complications. The prognosis of neurobrucellosis in children is usually good. PMID- 9733382 TI - Non-toxigenic Corynebacterium diphtheriae: two cases and review of the literature. AB - Non-toxigenic Corynebacterium diphtheriae infections are being reported with increasing frequency. We present two cases of C. diphtheriae endocarditis requiring early valve replacement. Both cases were complicated by cerebral embolic phenomena and pseudoaneurysm formation in lower limb arterial vessels. Non-toxigenic C. diphtheriae septicaemia must be excluded when 'diphtheroids' are isolated from blood cultures. PMID- 9733383 TI - Intra-articular empyema due to Staphylococcus caprae following arthroscopic cruciate ligament repair. AB - Staphylococcus caprae is a catalase-positive, coagulase-negative coccus that has been originally isolated from goat milk. We describe the first case of an intra articular empyema caused by S. caprae in an immunocompetent patient following arthroscopic cruciate ligament repair. The patient recovered completely after debridement and antibiotic therapy with cefazolin and amoxicillin, respectively. This case demonstrates that this organism may rarely cause serious nosocomial infections in immunocompetent adults. PMID- 9733384 TI - Anaerobiospirillum succiniciproducens septicaemia: important aspects of diagnosis and management. AB - Anaerobiospirillum succiniciproducens is a rare cause of septicaemia. A 63-year old woman with liver cirrhosis and a history of melaena developed A. succiniciprodocens septicaemia. She owned two pet dogs and a cat. Despite supportive management and antibiotic treatment supported by in vivo testing, the patient died. The characteristics identification and antimicrobial susceptibility of A. succiniciproducens are discussed and previous reported underlying disease reviewed. PMID- 9733385 TI - Management of brucella endocarditis of a prosthetic valve. AB - A case of chronic brucella endocarditis of a prosthetic valve is reported. The diagnosis of this infection was established by positive blood cultures and high brucella agglutination titre. The patient was successfully managed by combination of medical therapy (consisting of streptomycin, trimethoprim-sulphamethoxazole, rifampin and tetracycline) and surgery. PMID- 9733386 TI - Actinomyces pyogenes septic arthritis in a diabetic farmer. AB - We report a case of septic arthritis and osteomyelitis of the left ankle due to Actinomyces pyogenes in a diabetic farmer. Few confirmed human cases of A. pyogenes infection have been reported, partly because of inadequate identification of this bacterium. Bacteriological characteristics of the organism, which resembles Arcanobacterium haemolyticum, are described with a review of previous case reports. PMID- 9733387 TI - Oral histoplasmosis: a case report. AB - We report a case of biopsy-proven histoplasmosis in an 81-year-old man with mouth ulcers. The initial infection was probably contracted whilst he was a prisoner of war in Sumatra, and was reactivated during intercurrent illness with congestive cardiac failure and oral corticosteroid treatment for idiopathic thrombocytopaenic purpura. Of particular note is the latent period of 50 years, and a positive cytoplasmic antineutrophil cytoplasmic antibody (cANCA) titre. PMID- 9733388 TI - Haemophilus paraphrophilus; a rare cause of intracranial abscess. AB - We report a case of a 42-year-old man man who presented with neurological symptoms and was found to have an intracranial abscess. A stereotactic aspiration of the abscess yielded a pure growth of Haemophilus paraphrophilus. The patient responded to treatment with cefotaxime. We postulate the mechanism of infection in this patient. PMID- 9733389 TI - Listeria monocytogenes meningitis in a penicillin-allergic paediatric renal transplant patient. AB - Currently in many centres the extended spectrum cephalosporins (e.g. cefotaxime and ceftriaxone) are being used empirically for patients with suspected bacterial meningitis. We present a case of meningitis in a penicillin allergic paediatric renal transplant patient from whose cerebrospinal fluid (CSF) Listeria monocytogenes was cultured, despite four days of cefotaxime therapy. The patient was successfully treated with meropenem but required neuro-endoscopic intervention for hydrocephalus. PMID- 9733391 TI - Glycopeptide-induced vasculitis--cross-reactivity between vancomycin and teicoplanin. AB - Teicoplanin has been suggested for use in patients suffering complications from vancomycin. We describe two patients who developed a vasculitic rash whilst on vancomycin with recrudescence of the rash with subsequent teicoplanin therapy. PMID- 9733390 TI - Q fever in pregnancy: case report after a 2-year follow-up. AB - We report an acute Q fever case, a febrile syndrome, in the 14th week of pregnancy. Placental infection was documented by Coxiella burnetii culture. Newborn infection was ruled out on the basis of the absence of serological evidence after 2 years and on clinical normality. Serological diagnosis is reviewed here, as maternal serology was suggestive of chronic Q fever. The clinical progress, following extended observation, was consistent with acute infection. A QpDV plasmid, already described as being common to acute and chronic European cases, was detected. PMID- 9733392 TI - Jarisch-Herxheimer reaction complicating the treatment of chronic Q fever endocarditis: elevated TNFalpha and IL-6 serum levels. AB - Jarisch-Herxheimer reaction (J-HR) is an acute febrile reaction which may complicate the initiation of an effective treatment against infections due to intracellular micro-organisms. We report a case of J-HR complicating treatment of chronic Q fever endocarditis with demonstration of elevated serum cytokine concentrations. PMID- 9733393 TI - Can alprostadil improve liver failure in HIV-infected patients with severe acute viral hepatitis? AB - Liver failure with hepatic encephalopathy during an acute viral hepatitis carries a very high mortality. Liver transplantation is the usual treatment, but for poor candidates for transplantation only supportive therapy is available. Two patients with HIV infection developed an acute B hepatitis with liver insufficiency and hepatic encephalopathy. After an alprostadil infusion was begun they improved quickly and made a full recovery. This drug merits further investigation. PMID- 9733394 TI - Failure of anti-pneumococcal vaccine and prophylactic penicillin in a splenectomized patient. PMID- 9733395 TI - Acute meningitis caused by Streptococcus constellatus. PMID- 9733396 TI - Human fascioliasis: seasonal variations and female preponderance of complicated forms. PMID- 9733397 TI - Acute bacterial meningitis after dental fillings. PMID- 9733398 TI - Bilateral thrombosis of the internal jugular veins with spasmodic torticollis in a patient with acquired immunodeficiency syndrome and disseminated cytomegalovirus infection. PMID- 9733399 TI - Streptococcus mitis: urinary tract infection in a renal transplant patient. PMID- 9733400 TI - Improvement of seizure control by psychological methods in patients with intractable epilepsies. AB - Recent attempts to clarify the pathogenesis of pharmacoresistant epilepsies arrive at the conclusion that intractable epilepsies might be prevented by earlier, more effective pharmacotherapy. In this paper the problem of intractability is examined from a psychological point of view. Sixteen patients with intractable epilepsies were trained in techniques of self control (SC) in addition to ongoing pharmacological treatment. The SC training consisted of detailed self observation which aimed at identifying warning signals of a beginning seizure and seizure-provoking factors and the development of 'counter measures' (behavioural measures to interrupt a beginning seizure and to neutralize provoking factors). After SC training, all those patients who successfully managed to deal with their identified problems (strong psychic stress and/or poor intuitive SC abilities) achieved a significant improvement of seizure control: 68% obtained 80-100% reduction and 12% obtained 60-70% reduction of seizures. None changed for the worse. These findings suggested that psychological methods of seizure control can contribute to improving long standing intractable epilepsies. Offered early in the process of epilepsy they may even help to prevent the development of intractability. A new kind of polytherapy is proposed, consisting of a combination of pharmacological and SC therapy. PMID- 9733401 TI - Reading epilepsy: report of five new cases and further considerations on the pathophysiology. AB - Five new cases of reading epilepsy (RE) are reported. This is an epilepsy syndrome belonging to the group of idiopathic localization-related epilepsies. They all have some interesting features which contribute to the understanding of the pathomechanism and nosology of this specific type of reflex epilepsy. In our first patient the precipitating effect of texts in unknown languages depended upon phonematic intricacy. With our second case, changes of script within the text (Latin to Greek) increased the precipitating effect. The visual aura experiences reported in vague terms by some patients with RE may represent ictal dyslexia. For case three RE had been misdiagnosed as phobic neurosis. Even if a patient has a history of development dyslexia, and ictal dyslexia is a feature of the seizures, the onset of RE is not during primary school age but at puberty. Our fourth patient's manifestation factor of (late-onset) RE was a change of scriptural environment (Kyrillic to Latin) . Unilateral myocloni were observed with bilateral spike wave discharge in RE, and carbamazepine possibly increased the epileptic response in RE. With the co-occurrence of RE and juvenile myoclonic epilepsy in case five, the clinical features of both syndromes remained separate. All five patients responded well to treatment with valproic acid, and all confirmed that the syndrome has no tendency to deteriorate in long-term follow up. PMID- 9733402 TI - The Yelandur study: a community-based approach to epilepsy in rural South India- epidemiological aspects. AB - Data on the epidemiology of epilepsy in a rural community in a developing country would be of value in planning a decentralized management of this malady in its early stages commensurate with available local resources. A detailed screening instrument covering various seizure types was used by trained paramedical workers in a door-to-door survey of a population of 64,963 in rural South India. The prevalence period was from 1 April 1990 to 31 March 1991. The crude prevalence rate per 1000 for active epilepsy was 4.38 for males, 3.40 for females and 3.91 for both. The minimum and maximum prevalence rates, the latter computed from a validation sample, were 3.91 and 4.63 for active epilepsy; 0.28 and 0.77 for inactive epilepsy and 4.19 and 5.41 for life-time prevalence. In addition, corresponding figures for hot-water epilepsy, a type of reflex epilepsy peculiar to this area, were 2.49 and 2.99 for active phase; 0.35 and 0.85 for inactive phase and 2.85 and 3.83 for life-time prevalence. The incidence rate for epilepsy was 49.3 per 100,000, the same as in developed countries. These data do not support the concept that the prevalence of epilepsy in developing countries is twice that in the developed world. However, the role of local/regional variations should be borne in mind before extrapolating the figures to an entire country. PMID- 9733403 TI - Sudden unexpected death in epilepsy: is carbamazepine implicated? AB - Sudden unexpected death in epilepsy (SUDEP) has been recognised for centuries. The precise frequency of occurrence is not well defined. Education of medical professionals is needed, so that death certificates and coronial inquests may appropriately, correctly and consistently record SUDEP as the case of death. Correct identification will then allow further investigation of this misunderstood, and often ignored, epilepsy complication. SUDEP incidence may be increasing, either as a result of increased recognition, or possibly due to a real increase in incidence. All currently available antiepileptic drugs (AEDs) have been associated with SUDEP, and current opinion assumes that the relative proportion of patients suffering SUDEP is representative of average AED usage type for a particular time and locality, however, recently analysed data suggest a strong bias towards carbamazepine. A review of Cardiff Epilepsy Unit data shows that carbamazepine was disproportionately represented in patients suffering SUDEP. In this series, 11 of the 14 SUDEP patients were taking carbamazepine at the time of death. This was calculated as 79% of all patients, compared to average carbamazepine usage by all other Cardiff Epilepsy Unit patients of 38%. The data also indicate that one patient was not taking any drug therapy, and died during his first seizure, reducing the number of evaluable 'drug usage' patients to 13, and increasing the proportion taking carbamazepine at the time of death to 85%, (P < 0.01). Possible mechanisms include carbamazepine induced lengthening of the ECG Q-T interval combined with a mild pro-arrhythmic effect of epileptic seizure discharges, and consequent transient cardiac instability leading to arrhythmic death. Or alternatively, excessive post-seizure brainstem inhibition might result in blunting or transient abolition of central hypoxic and hypercarbic respiratory drive, with consequent post-ictal respiratory arrest, subsequent exacerbation of hypoxia, further cardiac destabilisation and death due to hypoxia/failed re-establishment of respiration and terminal cardiac arrhythmia. Current knowledge about SUDEP remains poor. Education is needed so that case ascertainment can be correctly documented. Delineation of the precise mechanisms involved should lead to definitive prevention strategies. Evaluation of carbamazepine as a significant causative factor in SUDEP is also needed. PMID- 9733404 TI - Long-term follow-up study of vigabatrin in pretreated children with West syndrome. AB - A multicentre, long-term, open-label, add-on study of vigabatrin was undertaken in 23 pretreated children with infantile spasms. After 3 months of vigabatrin therapy 11 of the 23 patients had become seizure-free. At this time two-thirds of these 11 children still received other antiepileptic drugs (AEDs) in addition to vigabatrin (mostly valproic acid and/or dexamethasone). After a mean follow-up time of 5 1/4 years (range: 4 1/4-6 1/2) 72% of 18 evaluable patients (two children died, three were lost to follow-up) revealed seizure freedom for at least 1 year. The mean duration of vigabatrin therapy had been 2 1/2 years (range: 2 weeks to 4 3/4 years). Two-thirds of the 18 children continued to take AEDs, three of them undergoing vigabatrin monotherapy. Relapses of infantile spasms had occurred in 14% of the children. The rate of vigabatrin side effects (10%) was low. At follow-up, the EEG of 13 and the 18 patients demonstrated focal or multifocal epileptic discharges. Fifty-five percent had developed another epilepsy (focal epilepsy, secondary generalized epilepsy or myoclonic-astatic epilepsy). With respect to mental functions, three children were normal or slightly retarded, four showed moderate retardation and 11 revealed severe or very severe retardation. This long-term result is comparable to that in ACTH studies with unselected patients. The conclusions are: (1) vigabatrin is an effective drug for the short-term and long-term treatment of refractory infantile spasms; (2) the relapse rate is low; (3) vigabatrin is well tolerated; (4) with respect to secondary epilepsies and mental functions the long-term outcome in these pretreated children is similar to that in earlier studies with ACTH or corticosteroids. PMID- 9733405 TI - Factors associated with the employment problems of people with established epilepsy. AB - The employment experiences of 245 respondents with epilepsy as their main diagnosis were examined as part of a study into the rehabilitation needs of an epilepsy outpatient clinic. It was found that 9% of the sample was unemployed and a further 16% were in receipt of a disability pension. Patients with seizures in remission were more likely to be employed and less likely to have experienced job problems, to feel limited by the epilepsy or to experience stigma. Job problems per se were experienced by 35% of the population. Of those with uncontrolled seizures, 50% had had job problems; however, 22% thought that their current employment situation had not been unduly influenced by epilepsy. It was those respondents who were younger or who were diagnosed early with epilepsy who were most likely to perceive their current situation as a result of having epilepsy. The survey suggests that unemployment is not the major problem it was once thought to be but that discrimination at work is a more serious problem which could lead to under-employment and restricted career development. Career planning should be instigated early and employment services should include information, practical advice and emotional support. PMID- 9733406 TI - Driving and epilepsy in Sri Lanka. AB - Regulations regarding driving for patients with epilepsy vary from country to country. They are well implemented in developed countries, but this is not the case in countries such as Sri Lanka. The aims of this study were to study characteristics of a cohort of patients with epilepsy who were driving or riding a vehicle at present, and study the attitudes of a representative sample of doctors, patients with epilepsy and the general population regarding aspects of driving by patients with epilepsy. Patients with epilepsy attending the medical clinics at the Colombo North General Hospital, Ragama, who were driving, were given a questionnaire and interviewed in order to assess their seizure characteristics. Another questionnaire was administered to epileptic patients visiting the clinics, a sample from the general population (relatives visiting in patients at the University Medical Unit selected randomly), doctors working at the General Hospital in Ragama and the Base Hospital in Negombo, and general practitioners in the Gampaha district, where these two hospitals are situated, which was designed to assess their views regarding driving by persons with epilepsy. Of the patients with epilepsy interviewed 24.8% were presently driving a vehicle, of them 51% were riding a motorcycle. The attitudes of the general public and patients to driving by epileptic patients were at opposite ends of the spectrum; 97% of the general public being opposed to driving by persons with epilepsy, while epileptics themselves being of the view that the rules should be lax. Doctors thought that there should be some regulations against driving by epileptic patients. These facts must be considered when setting implementable regulations regarding driving by epileptics in developing countries. PMID- 9733407 TI - An insight into children's and adolescents' experience of seizures and epilepsy. AB - A qualitative study was performed to investigate the individual experience of seizures and epilepsy in children and adolescents. Forty-one patients aged between 6 and 18 years old and affected with idiopathic epilepsy underwent one or more semi-structured interviews in a hospital day unit. Children aged 7 years or older could describe the experience of partial fits (in one case also of a presumably generalized fit). Seizures which occurred 6-12 months before had often been forgotten. Psychic involvement was reported in 90.3% cases, even when seizures had been classified as partial motor according to the parents' description. Social status and school achievement had no significant influence on the patient's ability to express his or her feelings, but some children had serious difficulty finding appropriate words to describe unfamiliar experiences; other patients used a simile, uncommon expressions or odd names to describe the fit. A poor relationship was found between seizure severity and patient's discomfort, and the image of the disease appeared independent of the experience of the seizures. As regards the epilepsy itself, patients seemed to suffer from generic problems rather than from specific concern about it, but some adolescents inserted their thoughts about the disease into reflections on their existential condition. PMID- 9733408 TI - The neurophysiological evaluation of nocturnal frontal lobe epilepsy. AB - The most reliable technique for the diagnosis of nocturnal frontal lobe epilepsy (NFLE) is nocturnal video-polysomnography, which is an expensive procedure and unavailable in many Departments of Neurology and Epileptology around the world. The aim of the present study was to evaluate the role of routine video-EEG and video-EEG after sleep deprivation, during the daytime, in the diagnosis of NFLE. We studied 23 patients complaining of repeated nocturnal motor attacks using a 3 level neurophysiological evaluation, including video-EEG when awake (level 1), video-EEG after sleep deprivation, during the daytime (level 2) and nocturnal video-polysomnography (level 3). All the patients had a normal video-EEG when awake. The video-EEG after sleep deprivation (level 2) allowed a diagnosis of NFLE in 52.2% of the patients, while the nocturnal video-polysomnography (level 3) allowed this diagnosis in 87.0% of the same sample. In the patients complaining of repeated nocturnal motor attacks, a video-EEG after sleep deprivation performed during the daytime, could be useful for diagnosis in about one half of cases. This methodology is routinely performed in many Departments of Neurology and Epileptology, and is much less binding and expensive than nocturnal video-polysomnography and so it could be important economically for the health service. PMID- 9733409 TI - Psychiatric disorder and cognitive function in children with epilepsy in Kerala, South India. AB - The cognitive and psychiatric associations of childhood epilepsy have not been studied in developing countries. Children with epilepsy were identified during a population-based epidemiological study of 1403 8- to 12-year-old children in Kerala, South India. They were compared with age-, sex- and social class-matched controls on measures of reading, vocabulary, non-verbal reasoning and school performance. In addition, psychiatric symptoms were measured using standard questionnaires and the presence or absence of psychiatric disorder was established by interview. Patients performed as well as controls on the non verbal test, but performed worse on tests of vocabulary and reading, suggesting a specific disadvantage in the area of education. Psychiatric disorder was more prevalent in the children with epilepsy. It was concluded that epilepsy in the population studied is accompanied by a significant burden of cognitive and psychiatric disorders which need recognition and adequate service provision. PMID- 9733410 TI - Primary reading epilepsy. AB - This is a report of a 23-year-old-man with primary reading epilepsy. He had had three generalized tonic-clonic seizures, each time beginning with visual illusions and occurring while reading a political science text in English with complicated words. He also described tightness and stiffness in the jaw and musculature of mastication for the past 2 years, which only lasted for a few seconds and only appeared while reading political science books in English. He noted that this sensation was associated with misreading of foreign and difficult words and disappeared when he stopped reading. This case report supports the view that difficult words and misreading are provocative factors evoking seizures in reading epilepsy. PMID- 9733411 TI - The perceived rehabilitation needs of a hospital-based outpatient sample of people with epilepsy. AB - A postal survey was carried out to identify the perceived epilepsy rehabilitation needs of a hospital-based outpatient population. A response rate of 70% resulted in 245 patients being surveyed. Data showed that 65% of the total sample wanted more rehabilitation assistance and that 27% required substantial contact with the rehabilitation services. The most common request was for more medical information both via written material and telephone contact with a specially trained epilepsy nurse. Approximately one in six patients would like to attend a course on how to live with epilepsy. Access to a psychologist was most often requested during the first year following diagnosis and demand for courses on how to live with epilepsy was highest in the second to fourth year following diagnosis. Requests for social worker assistance were associated with employment issues. Frequency of seizures, duration of epilepsy and age were significant variables in relation to demands for rehabilitation resources. The general conclusions are that (1) the minimum standards of a rehabilitation service should include greater access to medical information via a variety of authoritative sources; and (2) that team based resources are wanted by a substantial proportion of the population in relation to specific and definable problems, which would involve intensive input from psychologists and social workers. PMID- 9733412 TI - Postictal hemifacial purpura. AB - Non-traumatic stereotyped postictal purpura is rare. A 25-year-old woman presented with right facial, cheek and periorbital purpuric eruptions that occurred after secondarily generalized tonic-clonic seizures. The stereotyped, invariably right-sided facial skin eruption, which resolved in 48 hours, falsely raised concerns of spousal abuse. Possible pathophysiological mechanisms include: (a) valsalva-induced capillary hypertension with secondary purpura, (b) ictal corticolimbic stimulation of the autonomic nervous innervation of facial vasomotor structures, and (c) trigeminal-mediated local release of vasoactive substances. Although rare, such stereotyped patterns of purpura should be recognized to avoid incorrect attribution of spousal abuse. PMID- 9733413 TI - Symptomatic nocturnal frontal lobe epilepsy. PMID- 9733414 TI - Update: outbreak of influenza A infection--Alaska and the Yukon Territory, July August 1998. AB - On July 26, 1998, CDC and Health Canada, in cooperation with local public health authorities, began investigating reports of febrile respiratory illnesses and associated pneumonia among summer land and sea travelers to Alaska and the Yukon Territory (1). Epidemiologic and laboratory evidence has implicated influenza A virus as the etiologic agent of the outbreak. From June 11 through August 22, completed viral cultures of 101 (48%) of 209 nasopharyngeal specimens have yielded 26 influenza A isolates, four other respiratory viruses, and 71 negative results; results are pending for 108 additional specimens. Of the 26 influenza A isolates, five have been characterized at CDC; all have been identified as influenza A/Sydney/5/97 (H3N2)-like viruses, a strain included in the 1998-99 influenza vaccine. This report presents updated information about the outbreak and includes recommendations for influenza A prevention and control in this setting. PMID- 9733415 TI - Success in implementing Public Health Service guidelines to reduce perinatal transmission of HIV--Louisiana, Michigan, New Jersey, and South Carolina, 1993, 1995, and 1996. AB - In 1994, the Public Health Service (PHS) published guidelines for zidovudine (ZDV) use to reduce perinatal transmission of human immunodeficiency virus (HIV) (1), and in 1995 published guidelines for HIV counseling and voluntary testing of pregnant women (2). To directly assess the implementation of these guidelines and to identify barriers to the continued reduction of perinatal transmission, four states that conduct surveillance for HIV/acquired immunodeficiency syndrome (AIDS) (Louisiana, Michigan, New Jersey, and South Carolina) enhanced routine surveillance activities to conduct a population-based evaluation. This report summarizes the preliminary results of the evaluation, which identified 1) increases from 1993 to 1996 in the proportion of pregnant HIV-infected women in whom HIV infection was diagnosed before the birth of their child, 2) increases in the proportion of women offered ZDV and 3) lack of prenatal care as a critical obstacle to fully implementing the guidelines. PMID- 9733416 TI - Ciguatera fish poisoning--Texas, 1997. AB - On October 21, 1997, the Southeast Texas Poison Center was contacted by a local physician requesting information about treatment for crew members of a cargo ship docked in Freeport, Texas, who were ill with nausea, vomiting, diarrhea, and muscle weakness. This report summarizes an investigation of this outbreak by the Texas Department of Health (TDH), which indicated that 17 crew members experienced ciguatera fish poisoning resulting from eating a contaminated barracuda. PMID- 9733417 TI - Recruiting black men to a clinical trial to evaluate prostate cancer screening- Detroit, Michigan, 1998. AB - In 1998, an estimated 184,500 cases of prostate cancer will be diagnosed and approximately 39,200 men will die from this disease (1). Black men have higher prostate cancer incidence and mortality rates than white men (2). Representation of blacks in clinical trials that investigate the treatment of cancer is proportional to the burden of this disease in the black population (3). However, blacks have generally been underrepresented in clinical trials of preventive interventions (4). To determine the effect of socioeconomic status (SES) on the enrollment of black men in a trial that includes screening for prostate cancer, the African American Men (AAMEN) project in Detroit, Michigan, analyzed data from local recruitment efforts. This report summarizes preliminary results of this analysis, which indicate that SES was not an important factor in refusal to participate in the screening trial. PMID- 9733418 TI - Psychotherapeutic work with people with dementia: a review of the literature. AB - Until recently, psychotherapy and counselling techniques have rarely been used with people with dementia. However, the change in emphasis within dementia care towards a person-centred approach has meant that there is a growing clinical interest in their use. This paper reviews the published literature in this area which bears on psychotherapy assessment, intervention and evaluation. Although there is a developing clinical literature on intervention techniques drawn from all the main psychotherapeutic approaches, there has been little research into the effectiveness of this work and such research as does exist often uses methodologies that are inappropriate for such an early stage of clinical development. It is argued that clinical research should adopt case study or single-case designs that are more appropriate than group designs for evaluating new clinical developments. PMID- 9733419 TI - Adolescents who drop out of psychotherapy at a community-based psychotherapy centre: a preliminary investigation of the characteristics of early drop-outs, late drop-outs and those who continue treatment. AB - The present study examined the difference between young people who terminated treatment prematurely and who continued in treatment. One hundred and thirty-four young people (ages 12 to 24 years) who attended a community-based psychotherapy centre for psychoanalytic psychotherapy between 1 April 1993 and 31 March 1996 comprised the sample. It was predicted that drop-outs would consist of younger adolescents who were referred, who show a high score for externalizing problems such as aggression and delinquency and a low score for internalizing problems such as anxiety and depression. It was also predicted that continuers would be older, self-referred and show a high score for internalizing problems and a low score for externalizing problems. The results indicated significant differences between drop-outs and continuers: drop-outs were younger, had greater externalizing problems, school problems and presented with moderate to severe hyperkinetic or conduct disorder. Continuers were older, had fewer externalizing problems, were self-referred and were likely to be treated by supportive therapists. Since age was the most significant predictor of attendance, the sample was separated into younger adolescents and older adolescents and the same analyses repeated. In the younger group ethnic minority status, and being treated by a supportive therapist predicted continuing in treatment and a diagnosis of conduct disorder predicted premature termination. The clinical implications of the present findings for the delivery of psychotherapy services to young people are discussed. PMID- 9733420 TI - Follow-up study of British military hostages and their families held in Kuwait during the Gulf War. AB - On 2 August 1990 Iraq invaded Kuwait and held hostage all its inhabitants. Amongst those forced to stay were 71 British servicemen and their families who were held hostage for up to four and a half months. This study investigated the mental health status of this group of individuals at 6 and 18 months after the final hostage was released. Participants completed the Impact of Event Scale and the 28-item version of the General Health Questionnaire at both 6 and 18 months. In addition they completed a questionnaire regarding background factors, the dimensions of the trauma and the effects of their hostage experience. The Impact of Event Scale scores changed little over time whereas the General Health Questionnaire scores reduced significantly (p = .001) over the 12-month period suggesting that despite ongoing intrusive and avoidance phenomena levels of psychological distress did reduce. Those variables most strongly associated with a poor psychological outcome were witnessing physical violence and perceived deterioration in physical and mental health. Poor outcome at 6 months was strongly correlated with poor outcome at 18 months. PMID- 9733421 TI - Strategies for well-being in later life: a qualitative analysis. AB - A 70-year-old retired professional man, with some health problems but a high measured level of well-being, was interviewed for his perception of the sources of well-being. A grounded-theory analysis showed that he related well-being most strongly to the freedom to choose activities, and manage his own time. Other well being strategies within these domains were (i) reconstruing time, so that physical slow-down was assimilated to the positive values of leisure, patience and a steady pace of work, and (ii) social comparisons of self with others. Well being in the context of age-related symptoms was achieved explicitly through tenacious self-monitoring and self-management of physical and cognitive effort, and implicitly through minimizing or distancing of symptoms. The implicit strategies suggest a reorganization of subjectivity. The Discussion addresses the problems of inferring implicit strategies, and psychological processes specific to the experience of ageing. PMID- 9733422 TI - The contribution of adult attachment style to the adjustment to infertility. AB - This study examines the contribution of adult attachment style to the adjustment to infertility. Both husbands and wives of 80 infertile couples undergoing medical treatment completed the Attachment Style Scale, the Mental Health Inventory and the Dyadic Adjustment Scale. One year later, data were collected on whether women became pregnant. Diagnosis of male infertility was significantly more distressing than diagnosis of female infertility. Significant differences were found among attachment groups: secure persons, either men or women, reported more well-being, less distress and more dyadic adjustment than avoidant and anxious-ambivalent persons. Partners of secure persons also reported significantly higher levels of well-being and dyadic adjustment and significantly lower levels of distress than partners of anxious-ambivalent persons. However, these effects of attachment style were significant mainly when male infertility was diagnosed. Husbands' secure attachment made a significant positive contribution to pregnancy likelihood and this effect was mediated by adjustment measures. Results are discussed in terms of attachment theory. PMID- 9733423 TI - Hypochondriacal concerns, symptom reporting and secondary gain mechanisms. AB - Hypochondriacal concerns (HCs) and somatic symptom reporting (SSR) are associated. However, HCs are believed to be linked to the ego-defence coping strategy of avoiding help and SSR is believed to be linked to secondary gain. One hundred and twenty undergraduate students completed measures of HCs, SSR and the Desire for Control (DC) Scale. Subscales of the DC scale were used as indices of help avoidance and secondary gain. Both hierarchical multiple linear regression and LISREL structural modelling were used to control for the confound between HCs and SSR and explore the links with help avoidance and secondary gain. HCs were found to be primarily associated with an increased desire to strive for independence (avoid help) and levels of SSR were associated with the desire to have others make decisions (secondary gain). PMID- 9733424 TI - Can social cognitive models contribute to the effectiveness of HIV-preventive behavioural interventions? A brief review of the literature and a reply to Joffe (1996; 1997) and Fife-Schaw (1997) AB - A recent debate in the British Journal of Medical Psychology has considered the role of social cognitive models, such as the theory of reasoned action and the theory of planned behaviour, in understanding HIV-preventive behaviour. In this paper we clarify some of the assumptions involved in applications of social cognitive models. We briefly review available evidence on the capacity of such models to predict HIV-preventive sexual behaviour and outline a number of criteria for judging their predictive success. The importance of behavioural prediction for the development of effective HIV-preventive behavioural interventions is discussed and recent evaluations of interventions based on these models are reviewed. We conclude that the models are effective in predicting HIV preventive behaviours and provide empirically supported theoretical guidance on psychological changes likely to result in HIV-preventive behaviour change. In addition we argue that, to date, evaluations of theoretically specified interventions are encouraging. Further development and rigorous testing of HIV/AIDS interventions based on social cognitive models is recommended. PMID- 9733425 TI - Vulnerability factors in the anxiety disorders. AB - Groups of non-clinical individuals scoring high and low on a measure of trait anxiety were compared to a group of people with a history of clinical anxiety on measures of locus of control of behaviour and reported parental rearing practices (group size = 30 in each group). The Parental Bonding Instrument (Parker, Tulping & Brown, 1979) was used to assess parental overprotection and care. A scale relating to parental sensitization was also used in this study. It was hypothesized that the high-trait anxiety group would produce similar results to the anxiety-disorder group on locus of control and parental overprotection, and similar results to the low-trait anxiety group on parental care and parental sensitization. The hypotheses were, in most cases, supported and a relationship between trait anxiety and parental overprotection was suggested. The results also suggested that parental sensitization may be particular to anxiety disorders. PMID- 9733426 TI - The role of unrecognized ambivalence: the mirroring of family conflicts within the professional and legal network. AB - This paper raises some questions about the deleterious effects of unrecognized (unconscious) ambivalence running through child care systems, the Children Act and the adversarial legal system. It is postulated that unrecognized ambivalence may become one of the powerful unconscious dynamics contributing to the mirroring of the family conflicts within the professional network. A detailed clinical example, in which we acted as expert witnesses within a child psychiatry hospital team, is used to illustrate how the denial of ambivalence in a highly complex case, that of a large family involved in care proceedings, led to a state of polarization between the family and a team of professionals which was replicated and amplified within the wider welfare and legal system. It is our view that a state of polarization is likely to produce 'all or nothing decisions' which may not be in the best interest of both children and parents. It is argued that the adversarial nature of the legal process and some underlying assumptions in the Children Act, may contribute to ambivalence being unrecognized. Then it is suggested that ambivalence is inevitable both for professionals and families, particularly in highly complex cases, but with its recognition, a resolution may take place. The conclusions contain some suggestions to promote this process. PMID- 9733427 TI - A comparison of the long-term effectiveness of distraction and focusing in the treatment of auditory hallucinations. AB - Cognitive-behavioural interventions for patients experiencing neuroleptic resistant auditory hallucinations have fallen into two main categories: those which encourage distraction as a coping strategy, and those which encourage patients to focus on or expose themselves to their hallucinations. A 20-session distraction treatment was compared with an equal length focusing treatment for 19 patients who were experiencing chronic auditory hallucinations. Patients were followed-up for approximately 2 years. No differences were observed between the groups for outcome on symptom severity overall, although the focusers showed a greater belief that their voices were their own thoughts at the final follow-up point. When the two groups were combined, there was a significant reduction in the frequency of hallucinations and the disruption to life caused by them during treatment, although this was not maintained at follow-up. During treatment, there was a significant increase in self-esteem for focusers but a significant decrease for distracters. At 2-year follow-up, both focusers and distracters showed a reduction in self-esteem in comparison to the end of therapy. The results show no overwhelming advantage of one treatment over the other and confirm previous observations of the difficulty of treating hallucinations with cognitive behaviour therapy (CBT). However, there was some indication that CBT influenced some important clinical variables and further investigation is warranted. PMID- 9733428 TI - Molecular mechanism of embryo-endometrial adhesion in primates: a future task for implantation biologists. PMID- 9733429 TI - Role for cell surface oligosaccharide in cell-cell recognition during implantation. PMID- 9733430 TI - Human chromosome deletions in Yq11, AZF candidate genes and male infertility: history and update. AB - Human chromosome deletions in Yq11 seem to occur frequently as de novo mutation events in men with idiopathic azoospermia or severe oligozoospermia. However, the molecular extensions of these deletions are variable. They can be large and therefore visible under the microscope or small, not visible under the microscope, and containing the deletion of one or more DNA loci recently mapped in an apparently consecutive order along the Yq11 chromosome region. The results of 20 extensive microdeletion screening programmes have now corroborated the prevalence of the deletion of three non-overlapping DNA regions in proximal, middle and distal Yq11, which were designated earlier as AZFa, AZFb and AZFc. Deletions of single DNA loci were also reported, but as de novo and as polymorphic mutation events. Their clinical significance with regard to the men's infertility should therefore initially be handled with caution. Multiple Y genes expressed in human testis have now been mapped to each AZF region. At least one of them should be functional in human spermatogenesis and, if mutated, cause azoospermia. However, gene-specific mutations leading to the azoospermia phenotype have not yet been found for any of these AZF candidate genes. This might raise the question as to whether an AZF gene really exists in Yq11 or if the azoospermia phenotypes are only observed after deletion of a complete AZF region, after deletion of its complete gene content. PMID- 9733431 TI - The molecular mechanisms of oocyte maturation and early embryonic development are unveiling new insights into reproductive medicine. AB - The purpose of the present review is to outline the current understanding on the molecular mechanisms governing various stages of oocyte maturation, transition from maternal to embryonic control and the initial steps of pre-embryo development. The cytoplasmic and nuclear maturation of the oocyte during pre ovulatory development can be viewed as separate entities. Cytoplasmic maturation and the acquisition of stores of RNA and protein dominates oocyte development between the premordial and pre-ovulatory stages of development. Initiation of nuclear maturation is marked by the breakdown of the nuclear envelope, or germinal vesicle and is triggered by the midcycle luteinizing hormone peak. In vitro, this is associated with a decrease in the intracellular concentrations of cAMP. This and several subsequent steps of meiosis are controlled by the M-phase promoting factor (MPF). While the constituents of MPF, p34cdc2 kinase and B-type cyclin, are also present in mitotically dividing cells, in meiotically dividing oocytes the regulation of MPF activity differs. An oocyte-specific protein kinase, c-mos, plays an important role in up-regulating the activity of MPF at various stages of final oocyte maturation. Several lines of evidence suggest that the proper function of the c-mos-MPF system is associated with important features of the last stages of oocyte maturation such as the resumption of meiotic maturation, inhibition of DNA replication between meiosis I and II, and the maintenance of the oocyte at metaphase II arrest until it is fertilized. Eventually the destruction of c-mos and active MPF following fertilization allows the initiation of mitotic cell division in the pre-embryo. The very first cell divisions of the human pre-embryo are still under the control of maternally inherited mRNA and protein. Several lines of evidence suggest that in humans, zygotic gene expression is initiated between the 4- and 8-cell stages, after which the pre-embryo begins to utilize its own genes. Some of the first genes to be expressed in the human pre-embryo encode proteins that are associated with cell division, extracellular growth modulatory signals as well as factors associated with implantation. We acknowledge that most of the data presented comes from species other than human, therefore at present the full biological role of the proposed regulatory pathways and control mechanisms for human biology remains speculative. PMID- 9733432 TI - Germ cell apoptosis in men with complete and incomplete spermiogenesis failure. AB - Germ cell apoptosis was evaluated in 11 men suffering from nonobstructive azoospermia and enrolled in a spermatid conception programme. In six of these patients, round spermatids (Sa stage) were the most advanced spermatogenic cells recovered from testicular biopsy samples. This condition is referred to as complete spermiogenesis failure. In the remaining five men, a few late elongated spermatids (Sd stage) were unexpectedly found in the testicular biopsy samples on the day of treatment. This condition is referred to as incomplete spermiogenesis failure. Germ cell apoptosis in both groups of patients was examined by analysing cell smears prepared from mechanically disintegrated testicular tissues using terminal deoxyribonucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL), which detects apoptosis-specific DNA fragmentation, and annexin-V binding, detecting apoptosis-related translocation of plasma membrane phosphatidylserine to the membrane's outer surface. Both methods were combined, in double-fluorescence labelling preparations, with immunocytochemical detection of proacrosin, a specific germline marker. Patients with complete spermiogenesis failure had significantly higher frequencies of primary spermatocytes and round spermatids carrying the apoptosis-specific DNA damage in comparison with patients with incomplete spermiogenesis failure. Surprisingly, apoptosis-related phosphatidylserine externalization occurs rarely until the advanced stages of spermiogenesis. Since externalized phosphatidylserine is expected to be involved in the recognition of apoptotic cells by phagocytes, apoptotic spermatocytes and round spermatids may not be removed easily by phagocytosis. The high frequency of DNA damage in round spermatids from patients with complete spermiogenesis failure explains the low success rates of spermatid conception in these cases. The evaluation of apoptosis can help predict success rates of spermatid conception. PMID- 9733433 TI - Y chromosome microdeletions, in azoospermic or near-azoospermic subjects, are located in the AZFc (DAZ) subregion. AB - Submicroscopic deletions of the Y chromosome and polymorphisms of the androgen receptor (AR) gene in the X chromosome have been observed in men with defective spermatogenesis. To further define the subregions/genes in the Y chromosome causing male infertility and its relationship to polymorphisms of the AR polyglutamine tract, we screened the genomic DNA of 202 subfertile males and 101 healthy fertile controls of predominantly Chinese ethnic origin. Y microdeletions were examined with 16 sequence-tagged site (STS) probes, including the RBM and DAZ genes, spanning the AZFb and AZFc subregions of Yq11, and related to the size of trinucleotide repeat encoding the AR polyglutamine tract. Y microdeletions were detected and confirmed in three out of 44 (6.8%) of azoospermic and three out of 86 (3.5%) severely oligozoospermic patients. No deletions were detected in any of the patients with sperm counts of >0.5 x 10(6)/ml, nor in any of the 101 fertile controls. All six affected patients had almost contiguous Y microdeletions spanning the entire AZFc region including the DAZ gene. The AZFb region, containing the RBM1 gene, was intact in five of the six subjects. Y deletions were not found in those with long AR polyglutamine tracts. Our study, the first in a Chinese population, suggest a cause and effect relationship between Y microdeletions in the AZFc region (possibly DAZ), and azoospermia or near-azoospermia. Y microdeletions and long AR polyglutamine tracts appear to be independent contributors to male infertility. PMID- 9733434 TI - Occurrence of GABA and GABA receptors in human spermatozoa. AB - Gamma-aminobutyric acid (GABA) concentrations in seminal plasma and washed spermatozoa from normal donors were assessed by a sensitive radioreceptor assay, and were detectable in both fractions. Specific binding of [3H]-muscimol was shown to be dependent on protein concentration, temperature and incubation time. [3H]-muscimol specific binding to human sperm membranes was significantly inhibited by the GABA type A receptor (GABA(A)) antagonist, bicuculline, and by the GABA(A) agonists, muscimol and isoguvacine, but not by the GABA type B receptor (GABA(B)) agonist baclofen. Scatchard analysis of [3H]-muscimol binding yielded a linear plot consistent with a single population of binding sites with a dissociation constant in the low nanomolar range. Incubation with GABA at a high micromolar concentration for 3 h under capacitating conditions resulted in an increase in the percentage of spermatozoa showing hyperactivated motility as assessed by computerized motility analyser. However, low micromolar concentrations of the GABA(A) agonist, muscimol, were sufficient to significantly increase sperm hyperactivity. These results suggest that the effect of GABA on human sperm motility might be mediated through a specific GABA(A) receptor. PMID- 9733435 TI - Annexin V labelling and terminal transferase-mediated DNA end labelling (TUNEL) assay in human arrested embryos. AB - Confocal laser scanning microscopy was used to observe human arrested and fragmented preimplantation embryos obtained by in-vitro fertilization. Observation of the cellular actin cortex and chromatin showed a high frequency of embryos with blastomeres exhibiting two or more nuclei, while others had nuclei displaying chromatin condensation and fragmentation patterns. Many of the abnormal chromatin images could be due to the process of programmed cell death (apoptosis). The possible link between abnormalities of the blastomeres and apoptosis was investigated using two detection methods for cells undergoing apoptosis. Detection of phosphatidylserine exposure was performed using annexin V; the chromosomal breakdown preceding the nuclear collapse of apoptotic nuclei was tested using the terminal transferase-mediated DNA end labelling (TUNEL) assay. Annexin V staining was observed in all arrested and/or fragmented human embryos, but not in cryopreserved embryos which continued to develop normally after thawing. The TUNEL assay was positive in 30% (15/50) of arrested embryos, all of which had cytoplasmic fragments. In contrast, embryos showing regular size blastomeres without fragments were TUNEL negative. PMID- 9733436 TI - Detection of a novel splice variant of the hypoxanthine-guanine phosphoribosyl transferase gene in human oocytes and preimplantation embryos: implications for a RT-PCR-based preimplantation diagnosis of Lesch-Nyhan syndrome. AB - We have detected a novel splice variant of the hypoxanthine-guanine phosphoribosyl transferase (HPRT) gene in two human oocytes and four preimplantation embryos from the 4-cell to the 8-cell stage of development. The novel HPRT transcript lacks exons 4, 5 and 6 of the normal HPRT gene. The same parental origin for the two oocytes and two of the preimplantation embryos, in which the alternatively spliced transcript was detected, might suggest that the alternative splicing is influenced by genetic background. Mutations in the HPRT gene which cause alternative mRNA splicing are implicated in Lesch-Nyhan syndrome. However, the relatively high frequency of detection of this novel HPRT transcript described here (6/109 oocytes and preimplantation embryos) suggests that it is not involved in Lesch-Nyhan syndrome. It is probable that the alternative HPRT transcript is derived from the aberrant splicing of a small percentage of the total mRNA produced from normal HPRT alleles. The presence of this alternative transcript in human preimplantation embryos may complicate an reverse transcription-polymerase chain reaction-based preimplantation diagnosis of Lesch-Nyhan syndrome. PMID- 9733437 TI - Fluorescent PCR and automated fragment analysis for the clinical application of preimplantation genetic diagnosis of myotonic dystrophy (Steinert's disease). AB - Myotonic dystrophy (DM), or Steinert's disease, is an autosomal dominant disease characterized by myotonia, muscular weakness and atrophy, as well as lens opacities, cardiomyopathy and mild endocrine changes. The gene for DM located on 19q contains a triplet repeat at the 3' end of the gene. In DM patients, this repeat is found to be expanded. We have previously described a preimplantation genetic diagnosis (PGD) for DM using polymerase chain reaction (PCR) followed by conventional analysis on ethidium bromide-stained gels. The major drawback of this system was that allelic dropout occurred in >20% of the cells, leading to the loss of healthy embryos for transfer. To resolve this problem, we developed a PGD for DM using fluorescent PCR followed by fragment analysis on an automated DNA sequencer and made a comparison between the conventional PCR described earlier and fluorescent PCR, which turned out to be superior in accuracy and efficiency. Three PGD cycles were performed using fluorescent PCR and are described here. PMID- 9733438 TI - Possible involvement of very low density lipoproteins in steroidogenesis in the human ovary. AB - To determine whether human luteal cells can utilize very low density lipoproteins (VLDL)-carried cholesterol for steroidogenesis, we investigated the expression of VLDL receptor mRNA in human ovarian tissues and progesterone production by human luteinized granulosa cells after the addition of VLDL. The production of progesterone in the presence of human chorionic gonadotrophin (HCG) was increased significantly (P < 0.05) by VLDL (2479 +/- 1477 ng/10(5) cells, mean +/- SD, n = 6) and low density lipoproteins (LDL) (2726 +/- 1287), in comparison with the level in the absence of these lipoproteins (1350 +/- 739). Northern blot analysis revealed that the levels of expression of VLDL and LDL receptor mRNA in granulosa cells were almost equal to those in whole ovarian tissue. VLDL receptor mRNA was abundant in granulosa cells of preovulatory follicles and cells of the corpus luteum. Preovulatory thecal cells and stromal cells expressed lower amounts of VLDL receptor mRNA than granulosa cells of preovulatory follicles and cells of the corpus luteum. From the present study, it might be suggested that VLDL is utilized for steroidogenesis in human luteinized granulosa cells. PMID- 9733439 TI - Human large luteal cells in the menstrual cycle and early pregnancy express leukotriene A4 hydrolase. AB - Leukotriene (LT) A4 hydrolase (EC3.3.2.6) converts LTA4 to LTB4 which shows a chemotactic activity to leukocytes. To investigate the involvement of LTB4 in human corpus luteum (CL) function, the localization of LTA4 hydrolase in human ovarian tissues was examined by immunohistochemistry with a rabbit polyclonal antibody against LTA4 hydrolase. The enzyme was weakly expressed on granulosa cells of the follicles. After ovulation, the intensity of LTA4 hydrolase on large luteal cells increased and was highest in the midluteal phase. High expression was also observed in the CL of early pregnancy. In theca interna and small luteal cells, LTA4 hydrolase was weakly detected in all developmental stages. Immunoblot analysis demonstrated that the molecular mass of LTA4 hydrolase expressed in CL was 62 kDa, and confirmed that LTA4 hydrolase expression increased during CL formation and remained high in early pregnancy. The sequence encoding mRNA of LTA4 hydrolase, which was isolated from CL and amplified by polymerase chain reaction, was shown to be identical to the previously reported one. Immunocytochemistry showed that LTA4 hydrolase expression in cultured granulosa cells increased over 4 days in vitro and was enhanced by human chorionic gonadotrophin treatment. These expression profiles of LTA4 hydrolase suggest the involvement of LTB4 in luteal cell function during CL formation and early pregnancy. PMID- 9733440 TI - Expression of oestrogen receptor alpha and beta in cultured human ovarian surface epithelial cells. AB - Ovarian surface epithelial (OSE) cells participate in the formation of the ovarian cortex and are potential targets of oestrogen action. Oestrogens typically act through nuclear oestrogen receptors (ER) of which there are two known subtypes: ERalpha and ERbeta. In view of the potential importance of oestrogen as a local regulator of OSE cell function, we screened for ERalpha and ERbeta mRNA in primary OSE cell cultures by reverse transcriptase-polymerase chain reaction (RT-PCR) analysis, and used freshly isolated granulosa cells (GC) and granulosa-lutein cells (GLC) as positive controls. OSE cells, scraped from the ovarian surface of women undergoing laparotomy for benign gynaecological conditions, were cultured for up to 21 days to obtain enough cells for mRNA extraction. GC were obtained from spontaneously cyclic women undergoing total hysterectomy; while GLC were obtained from follicular aspirates of gonadotrophin stimulated in-vitro fertilization patients. Total RNA (1 microg) was reverse transcribed into single-stranded cDNA for PCR (30 cycles) using primers selected to give specific ERalpha and ERbeta products. The ERalpha and ERbeta PCR products, authenticated by cloning and sequencing, were both weakly detectable by Southern analysis in cultured OSE cells and readily detectable in GC and GLC. These results show that cultured human OSE express both ERalpha and ERbeta mRNA, consistent with a role for oestrogen in the regulation of OSE cell function in vivo. PMID- 9733441 TI - Differential expression and regulation of a human transgene, HLA-B27, in mouse placental and embryonic cell lines. AB - Unlike other somatic cells, human placental trophoblast cells do not express the highly polymorphic HLA-A and HLA-B human leukocyte major histocompatibility antigens that would stimulate maternal immunological rejection of the fetus. To investigate mechanisms underlying cell lineage-specific expression, cell lines were generated from homozygous matings of HLA-B27 transgenic mice. Trophoblast cell lines were generated from gestation day 10 placentas and fibroblasts were cultured from gestation day 13/14 embryos. Polymerase chain reaction (PCR) readily identified HLA-B DNA in transgenic trophoblastic cells but specific mRNA was of low abundance, being detectable by reverse transcriptase PCR but not by Northern blot hybridization. HLA-B-specific protein in/on the trophoblast cells was undetectable by cell enzyme-linked immunosorbent assay and the protein was not induced by exposing the trophoblastic cells to interferon-gamma (IFN-gamma). Restricted expression was specific for the HLA-B transgene and its antigen; IFN gamma-inducible endogenous H-2Db class I antigens were detectable on the trophoblast cells. In contrast to the trophoblastic cells, HLA-B27 transgenic fibroblasts expressed IFN-gamma-inducible HLA class I antigens as well as H-2Db antigens. Thus, the mechanism(s) regulating expression of the polymorphic HLA-B antigen in trophoblastic cells is gene-specific, IFN-gamma-resistant and operative at the level of transcription or immediate post-transcription. PMID- 9733442 TI - Host-microbe interaction in reactive arthritis: does HLA-B27 have a direct effect? PMID- 9733443 TI - Bacillus Calmette-Guerin induced aseptic arthritis: an experimental model of reactive arthritis. PMID- 9733444 TI - Adenovirus mediated gene delivery to the joints of guinea pigs. AB - OBJECTIVE: To clarify in vivo applicability of adenovirus mediated gene delivery to examine a gene therapy for human joint diseases. METHODS: We directly injected vectors harbouring beta-galactosidase gene and transforming growth factor (TGF) beta1 gene into the joints of Hartley guinea pigs. Expressions of delivered LacZ were examined by 5-bromo-4-chloro-3-indolyl-beta-D-galactoside staining and reverse transcription-polymerase chain reaction. The levels of TGF-beta1 that were delivered to the joint and then transferred to the joint fluid were assessed by ELISA. RESULTS: LacZ expression was observed in almost all synovial tissue samples and in chondrocytes on the surface of degenerated cartilage. In the other organs, expression of delivered genes was not observed. For 2 weeks following gene delivery TGF-beta1 levels in joint fluid were significantly higher than the levels in the controls for 2 weeks. CONCLUSION: Direct gene delivery into the joint cavity is feasible with the in vivo gene delivery method using adenovirus vector and would be clinically applicable. PMID- 9733445 TI - Osteonecrosis in the rheumatoid femoral head. AB - OBJECTIVE: To define the prevalence and pathological spectrum of femoral head osteonecrosis in patients with rheumatoid arthritis (RA) and to correlate its presence with disease related clinical and therapeutic factors. METHODS: A total of 545 primary total hip arthroplasties performed in 507 patients with RA were identified. A historical review of each patient's rheumatoid disease and treatment as well as pathological review of each femoral head specimen was performed. RESULTS: Osteonecrosis was identified in 66 specimens (12.1%) in one of 2 discrete forms. Thirty-two specimens (5.9%) contained classic subchondral avascular necrosis. Thirty-four specimens (6.2%) contained osteonecrosis in association with degenerative changes (within regions of sclerotic and eburnated subchondral bone), but not classic avascular necrosis. Remaining femoral head specimens were characterized by inflammatory arthritis (431 specimens) or degenerative joint disease (48 specimens). Corticosteroid therapy was used in 81% of patients with avascular necrosis and 68% with degenerative osteonecrosis. This was significantly greater prevalence than in patients without osteonecrosis (33%). Average daily prednisone dosage was 8 mg and no association between dosage and the presence of osteonecrosis was identified. No correlation between pathological findings and clinical disease severity was identified. In 5 of 27 specimens showing classic avascular necrosis and 11 of 34 containing degenerative osteonecrosis, no steroid treatment had been administered. CONCLUSION: Femoral head osteonecrosis is present in about 12% of patients with RA at hip arthroplasty, and occurs in 2 forms -- classic avascular necrosis and degenerative necrosis. Both forms are significantly associated with corticosteroid use. "Low dose" therapy does not protect patients against the development of osteonecrosis. Additionally, baseline prevalence of osteonecrosis of about 3% occurs in the absence of steroid use and may be related to the underlying inflammatory diseases. Despite its association with osteonecrosis the net effect of corticosteroid therapy on the natural history of rheumatoid hip disease remains unclear. PMID- 9733446 TI - Forefoot deformity, pain, and mobility in rheumatoid and nonarthritic subjects. AB - OBJECTIVE: To evaluate how painful metatarsal arthritis affects foot and ankle mechanics and mobility. METHODS: We studied 16 symptomatic forefeet in 10 patients with rheumatoid arthritis (RA) and compared them with 14 asymptomatic forefeet in 7 nonarthritic subjects. RA limbs with significant disease at other locations were excluded. We measured pain and deformity of the foot using a visual analog scale and a modified articular index. A video based 3 dimensional gait analysis system and force platform were used to collect data on subjects walking barefoot at a self-selected pace according to an established protocol. Mobility level was quantified using the Sickness Impact Profile (SIP) ambulation subscale. RESULTS: We observed considerable pain and deformity of the forefeet of RA subjects. During gait, motion and force measures revealed that RA subjects significantly (p < 0.005) delayed and reduced forefoot loading, which minimized use of the foot as a rigid level for push off. As a result, stride lengths were shorter and gait was slower compared to nonarthritic subjects. SIP scores revealed that these changes in gait resulted in moderate disability in RA subjects (p=0.05). CONCLUSION: Impairments of the forefoot due to RA include pain and deformity, which produce characteristic stance phase abnormalities in foot function, a slow walking speed, and moderate disability. PMID- 9733447 TI - The development of rheumatoid arthritis after recombinant hepatitis B vaccination. AB - OBJECTIVE: Hepatitis B vaccination has been associated with reactive arthritis and rarely rheumatoid arthritis (RA). We defined the clinical, serologic, and immunogenetic background of patients developing RA, soon after recombinant hepatitis B vaccination. METHODS: The clinical, serologic, and HLA antigens of a cluster of firefighters who developed arthritis after prophylactic recombinant hepatitis B vaccination (5 subjects), as well as a second group of sporadic cases of arthritis (6 patients) after hepatitis B vaccination are described. RESULTS: Ten of 11 patients fulfilled revised American College of Rheumatology criteria for RA. All cases had persistent arthritis for more than 6 months; at 48 months followup 2 cases no longer had inflammatory arthritis. Nine patients required disease modifying antirheumatic drugs. Five subjects were HLA-DR4 positive. HLA class II genes expressing the RA shared motif were identified in 9/11 patients genotyped for HLA-DRbeta1 and DQbeta1 alleles (0401, 0101, or 0404). All the firefighters shared the HLA-DRbeta1 allele 0301 and the DQbeta1 allele 0201, with which it is in linkage disequilibrium. CONCLUSION: These polymorphic residues in the binding site of the MHC class II molecules of the affected patients appear capable of binding some peptide sequences of the recombinant vaccine peptides they received and may be responsible for hepatitis B vaccine triggering development of RA in these cases. Recombinant hepatitis B vaccine may trigger the development of RA in MHC class II genetically susceptible individuals. PMID- 9733449 TI - Cross cultural adaptation and validation of the Chinese Health Assessment Questionnaire for use in rheumatoid arthritis. AB - OBJECTIVE: The Health Assessment Questionnaire - Disability Index (HAQ), used as a disability and outcome measurement in rheumatoid arthritis (RA), has been validated in several languages, but not in Chinese. Our aim was to validate the Chinese version of HAQ (Chinese-HAQ) to suit the needs of Chinese speaking patients with RA in an Asian setting. METHODS: The original HAQ was modified in the context of Chinese culture and translated into Chinese by 2 translators aware of the objective of the questionnaire. The Chinese HAQ was self-administered by 42 patients with RA during their routine followup visit and one week later. RESULTS: The test-retest reliability assessed using Spearman's correlation coefficient was 0.84. Between dimensions measured in the HAQ, the highest test retest reliability was observed for walking (Spearman correlation coefficient rs=0.80) and the lowest was for eating (rs=0.54). The internal consistency of the scale using Cronbach's alpha was high at 0.86. In terms of criterion validity, the Chinese-HAQ score was found to correlate well with American College of Rheumatology functional status (rs=0.501, p=0.01). The Chinese-HAQ scores also correlated well with markers of disease activity such as patient's perception of pain measured on a visual analog scale (rs=0.55, p < 0.001), grip strength in mm Hg (rs=-0.55. p < 0.001 ), and physician's assessment of disease activity (rs=0.59, p < 0.001). CONCLUSION: The Chinese HAQ is a reliable and valid instrument for studies measuring disability of patients with RA in Singapore. PMID- 9733448 TI - High dose versus low dose fludarabine in the treatment of patients with severe refractory rheumatoid arthritis. AB - OBJECTIVE: Fludarabine, a nucleoside analog that targets both resting and proliferating lymphocytes, is a promising drug for the treatment of autoimmune diseases. We conducted a 2 dose, open label clinical trial to evaluate the toxicity/safety of the fludarabine treatment and its clinical and immunological effects. METHODS: Twenty-six patients with severe rheumatoid arthritis (RA) refractory to treatment with at least one slow acting antirheumatic drug were treated with intravenous fludarabine [20 mg/m2 body surface area (n=12) or 30 mg/m2 body surface area (n=14) per day for 3 consecutive days] given monthly for 6 months. Second line agents with the exception of glucocorticoids were discontinued at least 4 weeks before study entry. Measurements included toxicity and tolerability monitored at monthly intervals: efficacy, by both a 50% reduction in tender or swollen joint count and American College of Rheumatology (ACR) criteria for 20% response; and phenotypic analysis of peripheral blood mononuclear cells and T cell functional assays. RESULTS: Using intention-to-treat analysis, 2 of 12 (17%) patients in the low dose and 7 of 14 (50%) in the high dose groups had 50% or greater reduction in tender and/or swollen joint count after 6 months of therapy compared to baseline (p=0.09). Two of 12 (17%) in the low dose group and 5 of 14 (36%) in the high dose group met ACR criteria for 20% improvement (p=0.28). No immediate toxicity was observed. Several infections occurred, including 4 episodes of limited Herpes zoster, which responded to standard therapy. Significant lymphopenia involving T and B cells was observed in all patients. Both naive (CD4+CD45RA+) and memory CD4+ T cells (CD4+CD45RO+) were reduced (naive > memory). No significant regeneration of naive T cells was observed, which may suggest limited thymic regenerative capacity. Fludarabine decreased the proliferative response of peripheral blood lymphocytes to mitogens, as well as the production of T cell (interleukin 2 and interferon-gamma) and monocyte derived (tumor necrosis factor-alpha and IL-10) cytokines. CONCLUSION: Fludarabine treatment of patients with severe, refractory RA resulted in significant lymphopenia, suppression of lymphocyte function, and clinical improvement in the high dose group. There was no immediate toxicity; however, several infections occurred. Controlled trials are needed to substantiate the clinical improvement observed in this open label trial. PMID- 9733450 TI - Why not use OSRA? A comparison of Overall Status in Rheumatoid Arthritis (RA) with ACR core set and other indices of disease activity in RA. AB - OBJECTIVE: The Overall Status in Rheumatoid Arthritis (OSRA) is a recently validated measure designed for routine immediate clinical use in patients with rheumatoid arthritis (RA). It is composed of demographic data, activity score (activity total), damage score (damage total), and drug treatment. We tested the hypothesis that this tool relates to existing measures and pooled indices of disease activity, including the SF-36. METHODS: Demographic information, OSRA, SF 36, and the ACR core set [inflammatory indicators (ESR, CRP), tender and swollen joints, visual analog scale for pain, Patient and Physician Global Assessment, and Health Assessment Questionnaire (HAQ)] were collected for 86 consecutive outpatients with RA who were starting or changing second-line therapy and again at 6 months. OSRA measures were examined for their relationship to all core set variables (SF-36, HAQ, Stoke Index, Disease Activity Score, and Mallya-Mace) using Spearman's rank correlation. OSRA was used to audit 246 consecutive outpatients with RA to determine its clinical utility. RESULTS: The median age was 58 years (range 29-82); median disease duration 63 mo (range 3-384); OSRA disease activity (mean 3.8, range 0-8) and damage (mean 2.7, range 0-7) scores were strongly associated with specific ACR core set and SF-36 measures, and all pooled indices examined. OSRA disease activity was significantly higher in outpatients in whom second-line therapy was changed. CONCLUSION: (1) The OSRA was highly correlated with HAQ and core set measures of disease activity: (2) the OSRA damage total was strongly associated with HAQ and correlated strongly with both duration and Larsen score; (3) OSRA scores also correlated well with specific SF-36 measures (activity total with Physical Functioning and Bodily Pain; damage total with Physical and Social Functioning); (4) OSRA shows good correlation with pooled indices that cannot be performed immediately in clinic; and (5) the OSRA activity score shows a strong association with clinical decisions made in the outpatient department. PMID- 9733451 TI - Smoking interferes with efficacy of antimalarial therapy in cutaneous lupus. AB - OBJECTIVE: There have been occasional reports of patients with refractory cutaneous lupus improving after cessation of cigarette smoking. It has been hypothesized that the effects of cigarette smoking on hepatic cytochrome P450 induction can alter the metabolism of antimalarials. Our objective was to determine the role of smoking in the efficacy of antimalarial therapy in cutaneous lupus. METHODS: A retrospective cohort study from the University of Toronto Lupus Clinic. Patients with either acute discoid or subacute cutaneous lupus (SACL) who received antimalarial therapy for their cutaneous lesions were selected. The smoking group consisted of regular smokers, while the nonsmoking group consisted of individuals who never smoked during the study period. The primary outcome measure was the complete resolution of the cutaneous lesion at 6 and 12 months of antimalarial therapy. Secondary outcome measures included the mean steroid dose and Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) score. Analysis included descriptive statistics and Fisher's exact test. RESULTS: Seventeen smokers (9 with discoid lupus, 5 SACL, 3 both) and 19 nonsmokers (11 discoid, 6 SACL, 2 both) were identified. The cutaneous eruption resolved completely in 3/17 smokers versus 9/17 nonsmokers after 6 months of antimalarial therapy (p < 0.035) and 3/16 smokers and 9/17 nonsmokers at 12 months (p < 0.046). There was no significant change in the mean steroid dose or SLEDAI in either group. CONCLUSION: Smoking appears to decrease the efficacy of antimalarial therapy in cutaneous lupus. The interaction between smoking and the efficacy of antimalarials in a variety of SLE presentations should be investigated further. PMID- 9733452 TI - Socioeconomic status and health in women with systemic lupus erythematosus. AB - OBJECTIVE: Health outcomes of patients with chronic illnesses are commonly worse in people of lower socioeconomic status (SES). We investigated psychosocial factors that may mediate the relationship between SES and measures of morbidity in women with systemic lupus erythematosus (SLE). METHODS: We collected information on SES, psychosocial factors, and health status in a cross sectional survey of 100 women with SLE. SES was rated using the Hollingshead Two-Factor Index, a weighted average of years of formal education and occupational prestige (higher Hollingshead Index=lower SES). Health status measures included the Health Assessment Questionnaire Disability Index (HAQ), the Systemic Lupus International Collaborating Clinics/American College of Rheumatology Damage Index (SLICC/ACR), the Systemic Lupus Activity Measure (SLAM), and the SLE Disease Activity Index (SLEDAI). Potential mediators consisted of 18 environmental, medical care, social, psychological, and behavioral factors. RESULTS: Patients with higher Hollingshead Indexes (lower SES) had more functional disability as measured by the HAQ (r=0.22: p=0.03) and more cumulative organ damage as measured by the SLICC/ACR Damage Index (r = 0.19; p=0.06). SES was not related to either the SLAM or SLEDAI. Significant univariate associations were present between the Hollingshead Index and 10 potential mediating variables: household crowding, insurance status, organizational barriers to medical care, depression, health locus of control by powerful others, SLE knowledge, social support, marital status, body mass index, and regular alcohol use. However, in multiple linear regression analyses, only 3 of these variables modified the relationship between Hollingshead Index and the HAQ: more severe depression scores, higher body mass index, and more restricted access to medical care. More severe depression and greater locus of control by powerful others tended to mediate the relationship between low SES and greater organ damage. CONCLUSION: SES is related to morbidity in women with SLE. There are identifiable and potentially modifiable mediators of this relationship. PMID- 9733453 TI - Clinical significance of immunoglobulin A antiphospholipid antibodies: possible association with skin manifestations and small vessel vasculitis. AB - OBJECTIVE: To clarify whether immunoglobulin A (IgA) antiphospholipid antibodies (aPL) are an independent risk factor for specific manifestations of collagen vascular diseases. METHODS: We determined IgG, IgM, and IgA anticardiolipin antibodies (aCL) and lupus anticoagulant (LAC) in 77 patients with various collagen diseases. Fifty-four patients who had positive results for either or both antibody classes were compared to 23 patients with systemic lupus erythematosus who had none of these antibodies. The association between the antibodies and clinical manifestations (thrombosis, fetal loss, thrombocytopenia, biological false positive test for syphilis, cutaneous manifestations, central nervous system involvement, and renal involvement) was analyzed. RESULTS: Of 54 patients with aPL, 33 showed significantly high levels of IgA aCL. Among them, IgA aCL coexisted with other aCL isotypes or LAC in 24 patients. The 9 patients with IgA aCL alone frequently had vasculitis associated manifestations, although thrombotic events and recurrent fetal loss were rare. Multivariate linear regression analysis showed that IgA aCL were independently associated with thrombocytopenia, skin ulcers, chilblain lupus, and vasculitis. There was also an association between IgM aCL and skin ulcers or chilblain lupus. CONCLUSION: Clinical manifestations of patients with IgA aCL differ from those of patients with IgG aCL. Determination of all 3 aCL isotypes and LAC is important to assess the risk of specific clinical manifestations in patients with aPL. PMID- 9733454 TI - Homocysteine, methylenetetrahydrofolate reductase polymorphism, antiphospholipid antibodies, and thromboembolic events in systemic lupus erythematosus: a retrospective cohort study. AB - OBJECTIVE: To examine the relationship between plasma homocysteine, 5,10 methylenetetrahydrofolate reductase (MTHFR) polymorphism, and the risk of arterial and/or venous thrombosis in systemic lupus erythematosus (SLE). METHODS: Plasma homocysteine and MTHFR polymorphism were determined in a retrospective cohort of 175 patients with SLE. Associations between homocysteine levels and antiphospholipid antibodies, renal function, and drug therapy were analyzed. RESULTS: Patients with arterial thrombosis had higher homocysteine concentration than nonthrombotic patients (20.6+/-10.2 vs 13.2+/-6.8 micromol/l; p < 0.001). The crude odds ratio for arterial thrombosis for the highest versus lowest quartile of homocysteine was 4.4 (95% CI 1.3-14.9). After adjustment for other risk factors the odds ratio remained significant (3.7; 95% CI 1.2-13.0). Increased plasma homocysteine was not associated with venous thrombosis. A significant correlation was found between homocysteine and creatinine (r=0.57, p < 0.0001) or the use of methotrexate. Homocysteine levels were higher in patients using corticosteroids and lower in patients using oral contraceptive drugs. This finding may be biased by impaired renal function in patients using corticosteroids and the advice to stop estrogen-containing contraceptives in female patients upon an episode of thrombosis. The distribution of MTHFR genotypes was 50% for homozygous wild type, 42% for heterozygous, and 8% for homozygous mutant, with a similar distribution among patients with or without a history of thrombosis. CONCLUSION: Raised levels of homocysteine are associated with arterial thrombosis in patients with SLE. Mutation in the MTHFR gene does not explain the raised levels, and renal failure is by far the most frequent cause. PMID- 9733455 TI - Anti-BB'-Sm antibodies, anticardiolipin antibodies, and thrombosis in systemic lupus erythematosus. AB - OBJECTIVE: Anticardiolipin antibodies (aCL) are associated with thrombosis in patients with systemic lupus erythematosus (SLE). Our aim was to determine whether there is an association between aCL, anti-Sm antibodies, and thrombosis in patients with SLE. METHODS: Sera from 153 patients with SLE were studied by ELISA, immunoblotting, and counter-immunoelectrophoresis (CIE) for anti-Sm antibodies, aCL and anti-dsDNA antibodies were detected by ELISA and radioimmunoassay, respectively. RESULTS: Anti-Sm antibodies were detected in 62 patients (40.5%) by ELISA and in 16 (10.4%) by CIE; IgG-anti-BB'-Sm in 44 (28.7%) by immunoblotting; IgG-aCL in 82 (53.5%), IgM-aCL in 44 (28.7%), and anti-dsDNA antibodies in 128 (83.6%). Anti-Sm and anti-dsDNA antibodies were significantly more frequent in patients with than in patients without aCL. Of the 89 patients with aCL, 36 (40.4%) showed IgG-anti-BB'-Sm antibodies (OR: 4.7; 95% CI: 2-10.5). Thrombosis was present in 20 (22.4%) SLE patients with aCL and in two (3.1 %) SLE patients without aCL (OR: 8.9; 95% CI: 2.4-31.8). Of the 22 patients with thrombosis, five (22.7%) had precipitating anti-Sm antibodies (OR: 3.2; 95% CI: 1.04-9.8) and 14 (63.6%) showed IgG-anti-BB'-Sm antibodies (OR: 5.8; 95% CI: 2.3 14). Anti-BB'-Sm and aCL were significantly more frequent in patients with anti dsDNA antibodies (32 and 62%) than in patients without these antibodies (12 and 36%) (OR: 3.4 and 2.9; 95% CI: 1.03-11.1 and 1.2-6.7, respectively). CONCLUSION: IgG-anti-BB'-Sm antibodies are associated with aCL, anti-dsDNA, and thrombosis in patients with SLE. Our findings suggest a possible association between anti-Sm, anti-dsDNA, and aCL responses in these patients. PMID- 9733456 TI - Polymyalgia rheumatica in biopsy proven giant cell arteritis does not constitute a different subset but differs from isolated polymyalgia rheumatica. AB - OBJECTIVE: To assess clinical and laboratory features that may be useful in differentiating isolated polymyalgia rheumatica (PMR) from PMR associated with biopsy proven giant cell arteritis (GCA); and in differentiating biopsy proven GCA associated with PMR from GCA without manifestations of PMR. METHODS: Clinical records of patients with PMR and biopsy proven GCA diagnosed at Hospital Xeral, Lugo, Spain from January 1987 through May 1997 were reviewed. Patients with a positive temporal artery biopsy were categorized into 2 different subgroups according to the presence or absence of associated PMR. The patients with biopsy proven GCA associated with PMR were compared with a group of patients with isolated PMR (not associated with GCA). RESULTS: From a total of 108 biopsy proven patients with GCA, 45 had associated PMR. Apart from a predominance of women and a longer delay to diagnosis, patients with PMR associated with GCA did not differ from the patients with GCA without PMR manifestations. In comparing patients with isolated PMR (n=117) with patients with PMR associated with GCA, we observed that PMR associated with GCA was a more severe disease, with significant abnormality in most laboratory variables, including constitutional syndrome, higher elevation of erythrocyte sedimentation rate and platelet counts, and lower values of hemoglobin. CONCLUSION: In both isolated PMR and PMR associated with GCA we observed a predominance of women. While there are no differences in the type of polymyalgia symptoms in patients with isolated PMR versus PMR associated with GCA, severe abnormalities associated with the inflammatory response in PMR may have prognostic value for more severe disease, which may be linked to the presence of GCA. PMID- 9733457 TI - Antibody response to Klebsiella pneumoniae 60 kDa protein in familial and sporadic ankylosing spondylitis: role of HLA-B27 and characterization as a GroEL like protein. AB - OBJECTIVE: To study the antibody response of HLA-B27+ patients with ankylosing spondylitis (AS) and their first degree relatives to the 60 kDa protein of Klebsiella pneumoniae and to characterize this protein. METHODS: Sera from 84 individuals were analyzed by ELISA to determine the titer of antibodies against the 60 kDa protein of K. pneumoniae. Subjects were divided into 3 categories: Group 1: 44 HLA-B27+ AS related individuals (35 patients, 9 healthy controls); Group 2: 28 healthy B27- AS related individuals; and Group 3: 12 healthy B27- non AS related subjects. The 60 kDa protein of K. pneumoniae was induced at 45 degrees C and purified by electroelution from sodium dodecyl sulfate polyacrylamide gel electrophoresis. It was characterized as a GroEL-like heat shock protein (HSP). The recognition of GroEL-like protein was confirmed by immunoblot of 2 dimension electrophoresis. The response to GroEL-like protein from other bacteria and the response to lipopolysaccharide (LPS) was also analyzed by immunoblot. RESULTS: HLA-B27+ individuals (Group 1), independent of their disease status, showed a significant higher response to the 60 kDa protein of K. pneumoniae than HLA-B27- subjects from Groups 2 and 3 (p < 0.0001). This protein was characterized as a HSP of the GroEL family and designated HSP60Kp. The GroEL of other enterobacteria as well as that of Mycobacterium leprae were recognized by HLA-B27+ individuals by immunoblot, whereas HLA-B27- individuals did not. LPS was not recognized by HLA-B27 positive or negative subjects. CONCLUSION: These findings suggest a relationship between HLA-B27 and the response to a GroEL-like protein that could have implications in AS. PMID- 9733458 TI - HLA-B27 does not affect invasion of arthritogenic bacteria into human cells. AB - OBJECTIVE: To investigate the effect of HLA-B27 expression on entry of Salmonella typhimurium and Yersinia enterocolitica into human cells. METHODS: We performed standard bacterial invasion assays with S. typhimurium and Y enterocolitica to analyze isogenic pairs of HeLa (epithelial), U937 (promonocyte), C1R (B lymphocyte), and Jurkat (T lymphocyte) human cell lines and their respective HLA B27 transfectants. Invasion of peripheral blood derived T lymphocytes, monocytes, and B lymphocytes/dendritic cell fraction (corresponding to peripheral blood cells depleted of monocytes and T lymphocytes) from patients with ankylosing spondylitis and healthy donors was also analyzed. The percentage of internalized bacteria was quantified, and the differences between HLA-B27 positive and negative samples were compared. RESULTS: The percentages of intracellular S. typhimurium and Y enterocolitica in HeLa, U937, and C1R with or without B27 were not statistically different (independent t test). We also found that the percentage of internalized bacteria did not differ significantly between HLA-B27 positive and negative samples in the different populations of peripheral blood derived cells. CONCLUSION: The presence of HLA-B27 on the surface of human cells does not alter the degree of bacterial invasion into either cultured human cell lines or peripheral blood derived human cells, and the influence of HLA-B27 expression on bacterial invasion should not be implicated in the pathogenesis of reactive arthritis related to Salmonella and Yersinia. PMID- 9733459 TI - Physiological characterization of mBSA antigen induced arthritis in the rat. I. Vascular leakiness and pannus growth. AB - OBJECTIVE: To study the temporal relation between vascular inflammatory activity and synovial hyperplasia during the development of methylated bovine serum albumin (mBSA) antigen induced arthritis (AIA) in the rat, and to correlate these variables to changes in knee diameter. The influence of a single dose of indomethacin and methotrexate (MTX) on these measures was also determined. METHODS: Vascular inflammatory activity was assessed as extravasation of radiolabelled albumin. Synovial hyperplasia was followed by measurements of the increases in wet and dry weight of the anterior part of the periarticular soft tissue and by routine histology. RESULTS: The vascular inflammation peaked on Day 3 after antigen challenge. The pannus weight increased at a slower pace, peaking on Day 7. No major difference between the sexes was found in these responses. Both variables were attenuated by MTX or indomethacin, suggesting a dependence between them. The water content of the pannus increased in tandem with the tissue growth but did not correlate to vascular leakiness, and is thus explained by the structural properties of the pannus rather than by the formation of inflammatory edema. In histological sections, ingrowth of pannus and destruction of cartilage was visible from Day 3 until the end of the experiment. CONCLUSION: Proliferative response follows the inflammatory vascular inflammation over time. The knee diameter, which is the most commonly used clinical measurement, seems mainly to be a reflection of the former variable. The effects of MTX and indomethacin suggest that the pannus formation is induced by the inflammatory activity in this model. PMID- 9733460 TI - Physiological characterization of mBSA antigen induced arthritis in the rat. II. Joint blood flow, glucose metabolism, and cell proliferation. AB - OBJECTIVE: Based on the hypothesis that blood flow in the inflamed joint is inadequate to maintain aerobic glycolysis, we sought to estimate the correlation between blood flow, glucose metabolism, and cellular proliferation rate in the arthritic joint. METHODS: Experiments were performed on rats with antigen induced arthritis (AIA). Regional blood flows (RBF) were measured with the microsphere technique, glucose metabolism by determination of [14C]2-deoxy-D-glucose (2-DG) uptake, and the proliferative response as the incorporation of [3H]-thymidine. RESULTS: In periarticular soft tissue of the arthritic knee the only significant change in the weight related RBF was an approximate 70% rise on Day 14 after arthritis onset. The RBF was lowest on Day 3 and the time course for the changes was inversely related to intensity of vascular inflammation. Weight related 2-DG uptake was more elevated than the RBF and peaked on Day 3. [3H]-thymidine incorporation in the soft tissue was only markedly enhanced on Day 3. Neither 2 DG nor [3H]-thymidine uptake was affected by treatment with methotrexate or indomethacin. In epiphyseal bone RBF was reduced on the first day of arthritis, but steadily increased thereafter. CONCLUSION: In AIA an intense vascular leakiness negatively affects the synovial blood. There is a marked enhancement of glucose metabolism, but only a minor part of this increase seems to be induced by increased cellular proliferation. PMID- 9733461 TI - Collagen induced arthritis: reversal by mercaptoethylguanidine, a novel antiinflammatory agent with a combined mechanism of action. AB - OBJECTIVE: We recently identified mercaptoethylguanidine (MEG) as an antiinflammatory agent with a combined mechanism of action. Its effects include inhibition of the inducible isoform of nitric oxide synthase (iNOS), scavenging peroxynitrite, a cytotoxic oxidant species produced from nitric oxide (NO) and superoxide, and inhibition of cyclooxygenase (COX). We investigate the effect of MEG in collagen induced arthritis (CIA). METHODS: Syngeneic LOU rats were immunized with native type II collagen on Day 0. After clinical signs of arthritis developed on Day 10, treatment with MEG was initiated (30 mg/kg ip tid) and continued until sacrifice on Day 28. Serum nitrite/nitrate was measured in control animals, at arthritis onset and 2 days after the start of MEG treatment. Clinical scores were obtained daily. At Day 28, radiographic scores were obtained, and joints were harvested for the measurement of mRNA for tumor necrosis factor-alpha (TNF-alpha), collagenase, and stromelysin. RESULTS: Serum nitrite/nitrate increased from 7.9+/-0.7 mM (baseline) to 13.5+/-2.6 at arthritis onset (p < 0.05). Within 48 h of MEG treatment, nitrite/nitrate levels fell to 7.2+/-1.1 (p < 0.05). By Day 28, clinical arthritis scores (measured on a scale of 0-8) were 7.1+/-0.6 in the vehicle group compared to 1.4+/-0.6 in the MEG treated group (p < 0.0001). Radiographic scores (scale 0-6) on Day 28 were reduced from 4.9+/-0.6 to 0.6+/-0.4 (p < 0.0002) by MEG treatment. MEG reduced the synovial expression of mRNA for TNF-alpha, collagenase, and stromelysin by 72, 67, and 52%, respectively. CONCLUSION: These data show that MEG has beneficial effects on established CIA. The mechanism of action may be related to inhibition of synovial iNOS expression or activity, inhibition of COX, scavenging of peroxynitrite, with subsequent inhibition of angiogenesis, metalloproteinase, and TNF-alpha expression. PMID- 9733462 TI - The role of T cells in polyethylene particulate induced inflammation. AB - OBJECTIVE: To investigate the role of T lymphocytes in ultra-high molecular weight polyethylene (UHMWPE) induced inflammation in joint arthroplasty. METHOD: We address the role of T cells in wear induced inflammation by injecting the knee joints of both immune competent rats and mice and severe combined immunodeficient (SCID) mice with UHMWPE. Histological and immunohistochemical analysis of the synovial tissues was compared. Interaction between human T cells and UHMWPE particles was examined in vitro using T cell activation assays. RESULTS: Histological and immunohistochemical analysis of the knees of the immune competent animals showed significant UHMWPE induced inflammation. In contrast, the tissue in the SCID mice knee joints showed very little inflammatory response to UHMWPE despite phagocytosis of the particulate. Since the SCID mice have no functional T or B lymphocytes, it is highly likely that the lack of inflammation in knee joints may be due to the absence of mouse T cells, as the infiltration of T cells into the joint tissue may enhance the inflammatory response to UHMWPE particles. T cell activation assays showed that T cells were not directly activated by UHMWPE particles and the nature of the interaction was not revealed from these experiments. CONCLUSIONS: Although T cells are not directly involved in UHMWPE particle induced inflammation, as shown by the T cell activation assays, the histological data from the mice studies clearly show differences in the amplitude of inflammation from animals with and without functional T cells. Our studies suggest that the T cells may enhance the inflammatory response due to a bystander effect. Since the macrophages upon ingestion of UHMWPE particles release several cytokines including tumor necrosis factor-alpha, interleukin 1, and IL-6, it is possible that T cells in the vicinity of these macrophages may become attracted to the knee joint and activated due to cytokine release. PMID- 9733463 TI - Is sonography performed by the rheumatologist as useful as arthrography executed by the radiologist for the assessment of full thickness rotator cuff tears? AB - OBJECTIVE: Sonography and arthrography are techniques used to detect rotator cuff tears (RCT). The diagnostic value of sonography executed by a rheumatologist versus that of arthrography performed by a radiologist for assessment of RCT was investigated among patients with unilateral chronic shoulder complaints without an underlying inflammatory rheumatic disease. METHODS: Forty-eight patients underwent sonography, arthrography, and surgical inspection, the results of surgical inspection being the gold standard. RESULTS: Sensitivity for assessment of a full thickness RCT was 0.86 for sonography and 0.77 for arthrography, specificity was 0.88 for sonography and 0.92 for arthrography. CONCLUSION: Compared to arthrography performed by the radiologist, sonography executed by the rheumatologist is as useful for assessment of full thickness rotator cuff tears. Sonography performed by the rheumatologist in combination with history taking and the physical examination during a single visit might change the routine daily practice of rheumatologists. PMID- 9733464 TI - Chemically modified non-antimicrobial tetracyclines inhibit activity of phospholipases A2. AB - OBJECTIVE: Tetracyclines have been recognized as useful agents for therapy of inflammatory arthritides. However, prolonged use of tetracyclines is limited by their detrimental antimicrobial properties. Recently, a group of chemically modified tetracyclines (CMT) devoid of antimicrobial properties has been synthesized. Some CMT were found to inhibit various matrix metalloproteinases (MMP). We reported previously that antimicrobial tetracyclines inhibit the activity of proinflammatory secretory group II phospholipase A2 (sPLA2). The objective of this study was to detect whether non-antimicrobial CMT also inhibit sPLA2 and other phospholipases A2. METHODS: Ten synthetic CMT were tested for inhibition of sPLA2 human and porcine PLA2, and Naja naja PLA2. PLA2 activity was assessed by radiolabeled Escherichia coli assay using standard and high calcium concentrations. RESULTS: Six of 10 CMT inhibited sPLA2 activity at concentrations close to or lower than 50 microg/ml. All 6 CMT had identical C1-3 and C10-12a positions in the 4-ringed nucleus of the tetracycline molecule. Calcium concentrations up to 20 mM did not eliminate the inhibitory activity of CMT. Inhibition of other PLA2 was induced by some CMT, all but one (CMT-9) belonging to the group of strong inhibitors of sPLA2. Thus, inhibition of PLA2 different from sPLA2 does not necessarily require identical C1-3/C10-12a residues. CONCLUSION: Since CMT, which inhibit proinflammatory sPLA2, are also inhibitors of some MMP, they may be useful for therapy of inflammatory diseases in which both MMP and sPLA2 are overexpressed. PMID- 9733465 TI - Histomorphometric and biochemical effect of various hyaluronans on early osteoarthritis. AB - OBJECTIVE: To evaluate, histomorphometrically and biochemically, different protective effects of clinically used hyaluronans (HA). METHODS: An experimental osteoarthritis (OA) model was applied to 132 mature NZW rabbits by resecting the unilateral anterior cruciate ligament (ACL). The other knee, where ACL remained intact, served as the control. We used native HA with different molecular weights, HA-50 (MW 5-7.3 x 10(5)), HA-80 (MW 8 x 10(5)), HA-360 (MW 3.6 x 10(6)), and crosslinked HA (HA-CL). HA were injected into the joint once a week for 5 weeks (HA-50, HA-80, HA-360) or 3 weeks (HA-CL) beginning 4 weeks after ACL transection. Histomorphometric and biochemical assessment was performed 9 weeks post-transection for both the HA treated and nontreated groups. RESULTS: In gross morphological observation, cartilage degeneration was suppressed in HA treated groups, and this effect was superior in the groups receiving either HA-80 or HA CL. Histomorphometric and biochemical analyses of the articular cartilage revealed similar results: the HA-80 and HA-CL groups showed no significant differences between the ACL transection and the control knees by histomorphometric variables, while the nontreated groups revealed significant degeneration. These evaluations were done in unblinded fashion. Biochemical analyses, including DNA synthesis in the synovium, also showed that articular cartilage and synovium in the HA-80 and HA-CL groups did not present significant changes compared to controls. CONCLUSION: In quantitative evaluation of this short term study using the OA model, native HA-80 and HA-CL presented a superior cartilage protective effect compared to the other native HA. PMID- 9733466 TI - The association of peripheral monocyte derived interleukin 1beta (IL-1beta), IL-1 receptor antagonist, and tumor necrosis factor-alpha with osteoarthritis in the elderly. AB - OBJECTIVE: To examine the association of peripheral blood mononuclear cell (PBMC) derived interleukin 1beta (IL-1beta), IL-1 receptor antagonist (IL-1Ra), tumor necrosis factor alpha (TNF-alpha), and radiographic osteoarthritis (OA) in the elderly. METHODS: A total of 703 subjects (436 women, 267 men, mean age 78.5+/ 4.5 yrs) had both knee and hand radiographs, and cytokines were measured during the 22nd biennial examination of the Framingham Cohort. PBMC derived IL-1beta , IL-1Ra, and TNF-alpha production was assessed using a non-cross reacting polyclonal radioimmunoassay. Knee OA was defined as a score of > 2 using a modified Kellgren and Lawrence scale. The presence of osteophytes and joint space narrowing were scored separately on a 0-3 scale, in which disease was defined a priori as a score > 0 for each feature. Sex-specific odds ratios were calculated for knee OA after adjusting for weight, history of knee injury, and use of estrogen and nonsteroidal antiinflammatory drugs. RESULT: No uniform associations were found for IL-1beta or IL-1Ra in men, or for TNF-alpha production and radiographic OA in either sex. We found possible associations for the highest levels of IL-1beta production and the presence of knee osteophytes [OR=2.0 (1.2 3.5)] and joint space narrowing [OR=1.7 (1.1-2.8)] in women. Our data suggested a possible protective effect for IL-1Ra production and hand OA in women [OR=0.6 (0.4-1.0)]. CONCLUSION: We found no consistent association of PBMC cytokine production and radiographic OA. However, women with the highest production of IL 1beta and IL-1Ra had respectively higher rates of knee OA and lower rates of hand OA than expected. PMID- 9733467 TI - Serum cholesterol and osteoarthritis. The baseline examination of the Ulm Osteoarthritis Study. AB - OBJECTIVE: To assess the association between serum cholesterol and osteoarthritis (OA). METHODS: OA patterns were studied in 809 patients with knee or hip joint replacement due to OA in 4 hospitals in southwest Germany. Participants had a standardized interview and examination. Radiographs of the contralateral joint as well as both hands and a blood sample were obtained. Serum cholesterol levels were divided into tertiles and hypercholesterolemia was defined as > or = 6.2 mmol/l or use of antihyperlipidemic drugs. According to the presence or absence of radiographic OA in the contralateral joint, participants were categorized as having bilateral or unilateral OA. If radiographic OA of different finger joints was present, participants were categorized as having generalized OA. Odds ratios and 95% confidence intervals for the association of serum cholesterol with OA patterns were calculated with logistic regression, adjusting for potential confounders. RESULTS: Eighty-five percent of participants with radiographs had bilateral OA and 26% generalized OA. No association was observed between hypercholesterolemia and bilateral OA. Hypercholesterolemia (OR 1.61; 95% CI 1.06 2.47) and high serum cholesterol levels (3rd versus 1st tertile: OR 1.73; 95% CI 1.02-2.92) were independently associated with generalized OA. This association was almost exclusively due to participants with knee OA. CONCLUSION: These data add to the evidence regarding the independent role of serum cholesterol as a systemic risk factor for OA. The discrepant associations observed for different OA patterns are likely due to the relative weight of other risk factors. PMID- 9733468 TI - Transdermal penetration of diclofenac after multiple epicutaneous administration. AB - OBJECTIVE: To test whether therapeutic diclofenac concentrations are attained in skeletal muscle tissue beneath the application site of an epicutaneously administered diclofenac foam formulation. METHODS: Diclofenac foam (5%) was administered epicutaneously at the thigh 80 mg/200 cm2 twice daily for a period of 7 days in healthy volunteers (n=12). On Day 8, 2 microdialysis probes were inserted into skeletal muscle tissue beneath the application site and an 80 mg dose was administered epicutaneously. Concentration versus time profiles in plasma and skeletal muscle were followed for 10 hours. RESULTS: Concentration versus time profiles were obtained for plasma and interstitial muscle fluid in all experiments. Mean Cmax in plasma was 18.75+/-4.97 ng/ml. Corresponding interstitial concentrations in skeletal muscle were significantly higher, 219.68+/-66.36 ng/ml (p=0.01). Plasma concentrations were not correlated to tissue concentrations (r=-0.08). CONCLUSION: There is significant direct penetration of diclofenac into skeletal muscle following multiple epicutaneous administration. However, the concentration attained in individual subjects is not predictable and may be strongly influenced by individual skin properties. PMID- 9733469 TI - Incidence and prevalence of juvenile chronic arthritis in East Berlin 1980-88. AB - OBJECTIVE: To estimate the incidence and prevalence rates of juvenile chronic arthritis (JCA). METHODS: The study population was children under 16 years of age living in the East Berlin area (part of the former German Democratic Republic). By admission order that was effective up to 1990, all children with symptoms of a rheumatic disease living in the East Berlin area had to be referred to the 2nd Children's Hospital at Berlin-Buch. This specific condition allowed us to ascertain cases from the clinical records and to calculate population rates. Based upon this data, the results of surveys with different methods of case ascertainment are compared. RESULTS: An incidence rate of 3.5 per 100,000 and a prevalence rate of 2.0 per 10,000 children were calculated. The frequency of JCA is higher for girls, with an incidence of 4.3 per 100,000 and a prevalence of 2.3 per 10,000. The figures for boys are 2.7 per 100,000 and 1.7 per 10,000, respectively. CONCLUSION: Because of the specific prerequisites, the population rates of prevalence and incidence that were based on clinical records can be regarded as valid in this study. Deviant results of other surveys can be explained by differences in the study design or in the diagnostic procedures used. PMID- 9733470 TI - Juvenile chronic arthritis in urban San Jose, Costa Rica: a 2 year prospective study. AB - OBJECTIVE: To find the incidence and prevalence of juvenile chronic arthritis (JCA) in the urban area of San Jose, Costa Rica. METHODS: During the year preceding our 2 year prospective, population based study, we conducted an educational program on JCA. The physicians caring for children < 16 years of age from all centers in the study area followed the program. They were asked to refer all cases of possible JCA according to EULAR criteria. The children were all evaluated at the National Children's Hospital. RESULTS: Of 189 children referred, 48 fulfilled EULAR criteria for JCA. The 2 year incidence rate for JCA was 13.7 per 100,000 children < 16 years old. This corresponds to an annual incidence per 100,000 children of 6.8 (95% CI 4.1-9.6). The incidence rate for pauciarticular onset JCA was 3.9 per 100,000. At the prevalence date, 122 cases of JCA were recorded, corresponding to a prevalence of 34.9 per 100,000 children < 16 years. When patients in remission were excluded, the prevalence was 31.4 per 100,000 (95% CI 25.5-37.2). The pauciarticular onset form was the most common, 71% of all prevalence cases. The highest incidence and prevalence were noted for pauciarticular girls with late onset JCA. No incidence peak was found in preschool age. The girl-to-boy ratio was 1.5/1. Antinuclear antibodies (ANA) were positive in only 7 cases (6.3%). IgM rheumatoid factor was found in 13 children (10.6%). Chronic iritis was observed in 4 cases, all of them ANA negative and older than 7 years of age at onset of arthritis. CONCLUSION: The incidence and prevalence observed were lower than those reported in other population based studies, but within the confidence intervals of their data. The incidence rate for pauciarticular JCA was significantly lower than that reported in other comparable studies. ANA positive pauciarticular preschool girls and associated uveitis were rarely encountered. PMID- 9733471 TI - Measuring disability in early juvenile rheumatoid arthritis: evaluation of a Norwegian version of the childhood Health Assessment Questionnaire. AB - OBJECTIVE: To assess the reliability, validity, and sensitivity to change of the Norwegian version of the childhood Health Assessment Questionnaire (CHAQ) and to examine the relationship between disability, disease severity, and psychosocial factors in patients with early juvenile rheumatoid arthritis (JRA). METHODS: Physical functioning was assessed by the CHAQ in 109 patients (median age 6.6 years, range 1.0-16.6) with JRA and a median of 4 months' (range 2-23) disease duration. Eighty-three patients were reassessed after a median of 6 months (range 3-21). Psychosocial functioning was assessed by the Child Behavior Checklist (n=39). RESULTS: The internal consistency of the CHAQ was good (Cronbach's alpha=0.83). The test-retest and parent-patient correlations were high [intraclass correlation coefficients 0.85 (n=18) and 0.75 (n=20), respectively, p < 0.001]. The CHAQ correlated moderately with number of tender, swollen and mobility restricted joints, morning stiffness, C-reactive protein, pain, and patients' and physicians' global assessments [correlation coefficients (r) ranging from 0.55 to 0.30, p < 0.01], but weakly with erythrocyte sedimentation rate (r=0.17, NS). The CHAQ also correlated with low levels of social competence (r=-0.49, p < 0.05) and high levels of internalizing behavior problems in the patients (r=0.43, p < 0.01) and low education levels of the mothers (r=-0.31, p < 0.01). Pain (beta 0.45, p < 0.001), number of swollen joints (beta 0.31, p < 0.001), and internalizing behavior problems (beta 0.45, p < 0.01) were predictors of disability. The median CHAQ changed from 0.25 to 0.00 (p < 0.05) in the 41 patients who improved, from 0.31 to 0.85 (p < 0.05) in the 18 patients whose condition was worse, and from 0.50 to 0.59 (NS) in the 24 patients whose condition was unchanged after 6 months. The effect size of the change was small (0.28) in those who improved and moderate (0.54) in those who became worse. CONCLUSION: The Norwegian version of the CHAQ is a reliable and valid instrument for measuring disability in children with early JRA. Pain, joint inflammation, and psychosocial factors are the most important correlates of disability and the CHAQ is sensitive to clinical change. PMID- 9733472 TI - Botulinum toxin increases tearing in patients with Sjogren's syndrome: a preliminary report. PMID- 9733473 TI - The use of standardized patients in needs assessment and CME. PMID- 9733474 TI - Video analysis of cytokine and cell adhesion molecule staining. PMID- 9733475 TI - Antibodies to the DEK protein in Kikuchi's disease. PMID- 9733476 TI - The increasing incidence of isolated congenital heart block in Finland. PMID- 9733477 TI - Autoimmune hemolytic anemia associated with eosinophilic fasciitis. PMID- 9733478 TI - Increased serum eosinophil cationic protein levels in familial Mediterranean fever. PMID- 9733479 TI - (Z,Z)-2,7-Bis(4-amidinobenzylidene)cycloheptan-1-one: identification of a highly active inhibitor of blood coagulation factor Xa. PMID- 9733481 TI - Conformationally constrained 1,3-diamino ketones: a series of potent inhibitors of the cysteine protease cathepsin K. PMID- 9733480 TI - Discovery of N-[2-[5-[Amino(imino)methyl]-2-hydroxyphenoxy]-3, 5-difluoro-6-[3 (4, 5-dihydro-1-methyl-1H-imidazol-2-yl)phenoxy]pyridin-4-yl]-N-methylgl y cine (ZK-807834): a potent, selective, and orally active inhibitor of the blood coagulation enzyme factor Xa. PMID- 9733482 TI - Discovery of potent, achiral matrix metalloproteinase inhibitors. PMID- 9733483 TI - Prodrugs of anthracyclines for use in antibody-directed enzyme prodrug therapy. AB - A series of new prodrugs of daunorubicin and doxorubicin which are candidates for antibody-directed enzyme prodrug therapy (ADEPT) is reported. These compounds (25a,b,c and 32a,b,c) have been designed to generate cytotoxic drugs after activation with beta-glucuronidase. As expected, recovery of the active drug was observed after enzymatic cleavage by Escherichia coli beta-glucuronidase as well as by a fusion protein which has been obtained from human beta-glucuronidase and humanized CEA-specific binding region. The six prodrugs are highly stable and are more than 100-fold less cytotoxic than doxorubicin against murine L1210 cell lines. The ortho-substituted phenyl carbamates 25a,b,c are better substrates for beta-glucuronidase than the corresponding para-substituted analogues. After taking into account additional factors such as stability in plasma and kinetics of enzymatic cleavage, we selected the o-nitro prodrug 25c for clinical trials. PMID- 9733484 TI - ATP-Citrate lyase as a target for hypolipidemic intervention. 2. Synthesis and evaluation of (3R,5S)-omega-substituted-3-carboxy-3, 5-dihydroxyalkanoic acids and their gamma-lactone prodrugs as inhibitors of the enzyme in vitro and in vivo. AB - A series of (3R,5S)-omega-substituted-3-carboxy-3, 5-dihydroxyalkanoic acids have been synthesized and evaluated as inhibitors of the recombinant human form of ATP citrate lyase. The best of these have Ki's in the 200-1000 nM range. As the corresponding thermodynamically favored gamma-lactone prodrugs, a number of compounds are able to inhibit cholesterol and fatty acid synthesis in HepG2 cells and reduce plasma triglyceride levels in vivo. The best of these, compound 77, is able to induce clear hypocholesterolemic and hypotriglyceridaemic responses when administered orally to rat and dog. These results provide evidence to support the hypothesis that compounds which inhibit ATP-citrate lyase have the potential to be a novel class of hypolipidemic agent, which possess combined hypocholesterolemic and hypotriglyceridemic activities. PMID- 9733485 TI - Classical/nonclassical hybrid cannabinoids: southern aliphatic chain functionalized C-6beta methyl, ethyl, and propyl analogues. AB - The stereoelectronic requirements for interaction of the southern aliphatic hydroxyl of cannabimimetic pharmacophores with the CB1 and CB2 receptors are explored. The stereoselective syntheses of three series of classical/nonclassical hybrid cannabinoids are described. These compounds were designed to investigate the importance of the southern aliphatic hydroxyl (SAH) pharmacophore for cannabimimetic activity. Variation in the chain length of the SAH moiety in these 6beta-(hydroxyalkyl)dihydrobenzopyran analogues, from 6beta-hydroxymethyl to 6beta-(omega-hydroxyethyl) and 6beta-(omega-hydroxypropyl), and the effects of replacing the hydroxyl functionality by hydride and iodide are reported. Our results indicate that the SAH pharmacophore has less pronounced effects than the C-3 aliphatic chain on cannabinoid activity. Furthermore, it appears that this southern molecular component is capable of interacting with two different subsites on the receptor and that the nature of this interaction is determined by the terminal substituent on the C-6beta alkyl group. One of the subsites can accommodate the relatively polar SAH pharmacophore, while the second subsite interacts with more hydrophobic C-6beta substituents and can accommodate large spherical pharmacophores separated by three methylene carbons from the tricyclic cannabinoid template. PMID- 9733486 TI - Derivation of a three-dimensional pharmacophore model of substance P antagonists bound to the neurokinin-1 receptor. AB - Constrained systematic search was used in an exhaustive conformational analysis of a structurally diverse set of substance P (SP) antagonists to identify a unique hypothesis for their bound conformation at the neurokinin-1 receptor. In this conformation, two aromatic groups essential for high affinity adopt a perpendicular or edge-on arrangement. This pharmacophore hypothesis for the receptor-bound conformation was used in a comparative molecular field analysis (CoMFA) of an expanded set of SP antagonists, and the predictive ability of the resulting three-dimensional quantitative structure-activity relationship (3D QSAR) was evaluated against a test set of SP antagonists different from those in the training set. This CoMFA model based on the Constrained Search alignment yielded significant cross-validated, conventional, and predictive r2 values equal to 0.70, 0.93, and 0.82, respectively. For comparison, the SP antagonists were forced into an alternative poorer alignment in which the two aromatic rings were parallel and then subjected to a CoMFA analysis. Both the parallel and perpendicular arrangements of the aromatic rings are seen in X-ray structures of SP antagonists and have been proposed as candidates for the receptor-bound conformation. The parallel (or stacked) conformation yielded a poorer correlation with a cross-validated r2 = 0.57, a conventional r2 = 0.90, and a predictive r2 = 0.78. Our results indicate that although both alignments could generate a reasonable CoMFA correlation, the stacked conformation is unlikely to be the receptor-bound conformation, as the covalent structure of the antagonists precludes a common geometry in which the aromatic rings are stacked. PMID- 9733487 TI - 2-[N-Acylamino(C1-C3)alkyl]indoles as MT1 melatonin receptor partial agonists, antagonists, and putative inverse agonists. AB - The synthesis of several novel indole melatonin analogues substituted at the 2 position with acylaminomethyl (8-11), acylaminoethyl (5a-k), or acylaminopropyl (13) side chains is reported. On the basis of a novel in vitro functional assay (specific binding of [35S]GTPgammaS), which can discriminate agonist from partial agonist, antagonist, and inverse agonist ligands, 5a,g, h,j and 13 were shown to be partial agonists, 5d,e and 8-11 competitive antagonists, and 5b,c,k putative inverse agonists. Binding and functional assays were performed on cloned human MT1 receptor. Structure-activity relationship considerations indicate that N-[1 aryl-2-(4-methoxy-1H-indol-2-yl)(C1-C2)alkyl]alkanamides represent a lead structure for this type of ligands. PMID- 9733488 TI - 2-Deoxy derivative is a partial agonist of the intracellular messenger inositol 3,4,5,6-tetrakisphosphate in the epithelial cell line T84. AB - We have synthesized the first deoxy analogues of myo-inositol 3,4,5, 6 tetrakisphosphate (1) [Ins(3,4,5,6)P4], rac-2-deoxy-myo-inositol 3, 4,5,6 tetrakisphosphate (rac-2), 2-deoxy-myo-inositol 1,4,5, 6-tetrakisphosphate (ent 2), and rac-1-deoxy-myo-inositol 3,4,5, 6-tetrakisphosphate (rac-3). In order to evaluate the binding properties of the three derivatives to the yet unidentified intracellular binding sites for Ins(3,4,5,6)P4, the analogues were converted to membrane-permeant derivatives. Starting with common inositol precursors, various forms of Barton-McCombie deoxygenation and classical protection/deprotection procedures yielded the desired precursors rac-1-O-butyryl-2-deoxy-myo-inositol (rac-12), ent-3-O-butyryl-2-deoxy-myo-inositol (ent-12), and rac-2-O-butyryl-1 deoxy-myo-inositol (rac-19), respectively. Phosphorylation and subsequent deprotection yielded rac-2, ent-2, and rac-3. Alternatively, phosphorylation followed by alkylation with acetoxymethyl bromide gave the membrane-permeant derivatives 1-O-butyryl-2-deoxy-myo-inositol 3,4,5,6-tetrakisphosphate octakis(acetoxymethyl) ester (rac-5), 3-O-butyryl-2-deoxy-myo-inositol 1,4,5,6 tetrakisphosphate octakis(acetoxymethyl) ester (ent-5), and 2-O-butyryl-1-deoxy myo-inositol 3,4,5,6-tetrakisphosphate octakis(acetoxymethyl) ester (rac-6), respectively. We examined the potency of the membrane-permeant deoxy derivatives in inhibition of calcium-mediated chloride secretion (CaMCS) in intact T84 cells. Compared to the 1,2-di-O-butyryl-myo-inositol 3,4,5, 6-tetrakisphosphate octakis(acetoxymethyl) ester (4), the membrane-permeant derivative of Ins(3,4,5,6)P4 (1), the 2-deoxy derivative (rac-5) exhibited a slightly weaker inhibitory effect, while the enantiomerically pure 2-deoxy-Ins(1,4,5,6)P4 (ent-5) and the 1-deoxy derivative (rac-6) were inactive. As expected, the effect was stereoselective. Thus, the 1-hydroxyl group is apparently essential for binding and the inhibitory effect of Ins(3,4,5,6)P4 on chloride secretion, whereas the 2 hydroxyl group plays a less important role. PMID- 9733489 TI - Synthesis and antitumor activity of 4-aminomethylthioxanthenone and 5 aminomethylbenzothiopyranoindazole derivatives. AB - Two new series of antitumor agents, 4-aminomethylthioxanthenones (6-50) and 5 aminomethylbenzothiopyranoindazoles (51-61), are described and compared. Nearly all members of both series display excellent in vivo activity versus murine pancreatic adenocarcinoma 03 (Panc03) although there is little to distinguish the two series from each other. In both series there is no discernible relationship between structure and in vivo efficacy. Selected analogues were evaluated in vitro; all were observed to have moderate to strong DNA binding via intercalation. However, varying degrees of in vitro P388 cytotoxicity and topoisomerase II inhibition were seen. In general, those molecules which exhibited strong topoisomerase II inhibition were significantly more cytotoxic than those which did not. In both series, those derivatives (48-50, 60, and 61) having a phenolic hydroxy substitution exhibited the most potent P388 cytotoxicity and topoisomerase II inhibition. PMID- 9733490 TI - Biologically active oligodeoxyribonucleotides. 5. 5'-End-substituted d(TGGGAG) possesses anti-human immunodeficiency virus type 1 activity by forming a G quadruplex structure. AB - A series of hexadeoxyribonucleotides (6-mers), d(TGGGAG), substituted with a variety of aromatic groups at the 5'-end were synthesized and tested for anti human immunodeficiency virus type 1 (HIV-1) activity. While unmodified d(TGGGAG) (31) had no anti-HIV-1 activity, compound 23 with a 3,4-di(benzyloxy)benzyl (DBB) group at the 5'-end potently inhibited the HIV-1IIIB-induced cytopathicity of MT 4 cells in vitro (IC50 = 0.37 microM) without cytotoxicity up to 40 microM. A thermal denaturation study on the 5'-end-substituted 6-mers by means of the circular dichroism (CD) spectra demonstrated that the aromatic substituent attached at the 5'-end of the 6-mer strongly enhanced the formation of a parallel helical structure consisting of four strands (quadruplex). On the contrary, compound 36, in which one of the guanosines of 23 was replaced by a thymidine, did not form a quadruplex, thus exhibiting no anti-HIV-1 activity. Moreover, both compound 15, with a tert-butyldiphenylsilyl group solely at its 3'-end, and compound 21, with a relatively small substituent, a benzyl group, at the 5'-end, formed quadruplexes but had no anti-HIV-1 activity. These findings led us to the conclusion that both the quadruplex structure and the aromatic substituent with adequate size at the 5'-end are crucial for the interaction of the 5'-end substituted 6-mers with the V3 loop as well as the CD4 binding site on viral gp120, resulting in anti-HIV-1 activity. PMID- 9733491 TI - Rational design, synthesis, and X-ray structure of selective noncovalent thrombin inhibitors. AB - We have designed, synthesized, and tested in vitro a novel class of noncovalent thrombin inhibitors. The main feature of these inhibitors is a 6,5-fused bicyclic core structure that fills the S2 pocket of the active site of thrombin. The bicycle introduces conformational constraint into the ligand and locks the Xaa Pro amide bond into the desired trans configuration. Among the known ring systems, we selected by molecular modeling the 7-thiaindolizidinones (BTD) as our basic template. The influence of several structural features was analyzed: the length of the argininal side chain, the stereochemistry at C6, and the importance of making optimal use of the S3 pocket. Finally, an X-ray crystal structure of inhibitor 15 bound to thrombin was obtained at a resolution of 2.3 A. These designed thrombin inhibitors, which were prepared by an efficient synthesis, showed high selectivity over trypsin and other serine proteases. Further derivation based on the information obtained by X-ray crystallography should certainly allow to improve the potency. PMID- 9733492 TI - 2-Iminopyrrolidines as potent and selective inhibitors of human inducible nitric oxide synthase. AB - A series of substituted 2-iminopyrrolidines has been prepared and shown to be potent and selective inhibitors of the human inducible nitric oxide synthase (hiNOS) isoform versus the human endothelial nitric oxide synthase (heNOS) and the human neuronal nitric oxide synthase (hnNOS). Simple substitutions at the 3-, 4-, or 5-position afforded more potent analogues than the parent 2 iminopyrrolidine 1. The effect of ring substitutions on both potency and selectivity for the different NOS isoforms is described. Substitution at the 4- and 5-positions of the 2-iminopyrrolidine yielded both potent and selective inhibitors of hiNOS. In particular, (+)-cis-4-methyl-5-pentylpyrrolidin-2-imine, monohydrochloride (20), displayed potent inhibition of hiNOS (IC50 = 0.25 microM) and selectivities of 897 (heNOS IC50/hiNOS IC50) and 13 (hnNOS IC50/hiNOS IC50). Example 20 was shown to be an efficacious inhibitor of NO production in the mouse endotoxin assay. Furthermore, 20 displayed in vivo selectivity, versus heNOS isoform, by not elevating blood pressure at multiples of the effective dose in the mouse. PMID- 9733493 TI - Reactions of melatonin and related indoles with free radicals: a computational study. AB - Melatonin is being increasingly promoted as a therapeutic agent for the treatment of jet lag and insomnia and has been recently suggested to act as an efficient free-radical scavenger. In the present work, its mechanisms of action for scavenging hydroxyl radicals have been investigated using semiempirical AM1 and density functional theory (DFT) computational tools. Two different reactions were proposed as follows: one involving the abstraction of an indolic hydrogen to yield a neutral radical and another involving the addition of the hydroxyl radical to the indolic moiety. Our results show that, from a thermodynamical standpoint, melatonin may directly scavenge hydroxyl radicals both in vacuum and in aqueous solution. The structural requirements for free-radical-trapping ability have been examined comparing melatonin with related indoles. Computational data suggest that 5-methoxy and N-acetyl groups of melatonin do not significantly affect its thermodynamical capacity of free-radical trapping. The present results support experimental data on the potential of melatonin as a physiological or pharmacological antioxidant agent. PMID- 9733494 TI - 2-, 5-, and 6-Halo-3-(2(S)-azetidinylmethoxy)pyridines: synthesis, affinity for nicotinic acetylcholine receptors, and molecular modeling. AB - 3-(2(S)-Azetidinylmethoxy)pyridine (A-85380) has been identified recently as a ligand with high affinity for nicotinic acetylcholine receptors (nAChRs). Here we report the synthesis and in vitro nAChR binding of a series of 10 pyridine modified analogues of A-85380. The novel compounds feature a halogen substituent at position 2, 5, or 6 of the 3-pyridyl fragment. Those with the substituents at position 5 or 6, as well as the 2-fluoro analogue, possess subnanomolar affinity for nAChRs in membranes from rat brain. For these ligands, Ki values range from 11 to 210 pM, as measured by competition with (+/-)-[3H]epibatidine. In contrast, 2-chloro, 2-bromo, and 2-iodo analogues exhibit substantially lower affinity. AM1 quantum chemical calculations demonstrate that the bulky substituents at position 2 cause notable changes in the molecular geometry. The high-affinity members of the series and (+)-epibatidine display a tight fit superposition of low-energy stable conformers. The new ligands with high affinity for nAChRs may be of interest as pharmacological probes, potential medications, and candidates for developing radiohalogenated tracers to study nAChRs. PMID- 9733495 TI - A new series of highly potent growth hormone-releasing peptides derived from ipamorelin. AB - A new series of GH secretagogues derived from ipamorelin is described. In an attempt to obtain oral bioavailability, by reducing the size and the number of potential hydrogen-bonding sites of the compounds, a strategy using the peptidomimetic fragment 3-(aminomethyl)benzoic acid and sequential backbone N methylations was applied. Several compounds from this series release GH with high in vitro potency and efficacy in a rat pituitary cell assay and high in vivo potency and efficacy in anesthetized rats. The tetrapeptide NNC 26-0235 (3 (aminomethyl)benzoyl-D-2Nal-N-Me-D-Phe-Lys-NH2) shows, following iv administration, comparable in vivo potency to ipamorelin, GHRP-2, and GHRP-6 with an ED50 in swine at 2 nmol/kg. NNC 26-0235 demonstrated a 10% oral bioavailability in dogs, and NNC 26-0235 and ipamorelin were able to increase basal GH level by more than 10-fold after oral administration of a dose of 1.8 and 2.7 mg/kg, respectively. The tripeptide NNC 26-0323 (3-(aminomethyl)benzoic acid-N-Me-D-2Nal-N-Me-D-Phe-ol) which showed moderate in vitro potency but lacked in vivo potency demonstrated a 20% oral bioavailability in rats. PMID- 9733496 TI - Novel orally active growth hormone secretagogues. AB - A novel class of growth hormone-releasing compounds with a molecular weight in the range from 500 to 650 has been discovered. The aim of this study was to obtain growth hormone secretagogues with oral bioavailability. By a rational approach we were able to reduce the size of the lead compound ipamorelin (4) and simultaneously to reduce hydrogen-bonding potential by incorporation of backbone isosters while retaining in vivo potency in swine. A rat pituitary assay was used for screening of all compounds and to evaluate which compounds should be tested further for in vivo potency in swine and oral bioavailability, fpo, in dogs. Most of the tested compounds had fpo in the range of 10-55%. In vivo potency in swine after iv dosing is reported, and ED50 was found to be 30 nmol/kg of body weight for the most potent compound. PMID- 9733497 TI - Synthesis and biological evaluation of 2-acyl analogues of paclitaxel (Taxol). AB - The anticancer drug paclitaxel (Taxol) has been converted to a large number of 2 debenzoyl-2-aroyl derivatives by three different methods. The bioactivities of the resulting analogues were determined in both tubulin polymerization and cytotoxicity assays, and several analogues with enhanced activity as compared with paclitaxel were discovered. Correlation of cytotoxicity in three cell lines with tubulin polymerization activity showed reasonable agreement. Among the cell lines examined, the closest correlation with antitubulin activity was observed with a human ovarian carcinoma cell line. PMID- 9733499 TI - Bicyclic acylguanidine Na+/H+ antiporter inhibitors. AB - Blockade of the Na+/H+ exchange has been shown to diminish the serious consequences of myocardial ischemia. The aim of this investigation was to alter the structure of the common benzoylguanidine NHE inhibitors in such a way that the 3-methylsulfonyl and 4-alkyl group form a ring. The new benz-fused five-, six , and seven-membered ring sulfones were prepared by internal Heck reaction. Benz fused five-membered ring sulfones could also be prepared by internal aldol-type condensation using ketones or nitriles as acceptor groups. In the final step, the carboxyl groups were converted to acylguanidines preferentially by guanidine treatment of the esters or acid chlorides. The compounds were tested as their methanesulfonate salts. The inhibition of the Na+/H+ antiport activity was determined by observing the uptake of 22Na+ into acidified rabbit erythrocytes. Additionally, the inhibition of the antiport activity was assessed also by the platelet swelling assay (PSA), in which the swelling of human platelets was induced by the incubation in the presence of a weak organic acid. On average, the IC50 values in the PSA turned out to be about 10-fold higher than in the erythrocyte assay primarily due to a higher Na+ concentration in the PSA; however, the order of the compounds' potency was not substantially altered. The new compounds were found to be highly active with peak values ranging within the cariporide and EMD 96785 standards. PMID- 9733498 TI - Piperazinyl oxazolidinone antibacterial agents containing a pyridine, diazene, or triazene heteroaromatic ring. AB - Oxazolidinones are a novel class of synthetic antibacterial agents active against gram-positive organisms including methicillin-resistant Staphylococcus aureus as well as selected anaerobic organisms. Important representatives of this class include the morpholine derivative linezolid 2, which is currently in phase III clinical trials, and the piperazine derivative eperezolid 3. As part of an investigation of the structure-activity relationships of structurally related oxazolidinones, we have prepared and evaluated the antibacterial properties of a series of piperazinyl oxazolidinones in which the distal nitrogen of the piperazinyl ring is substituted with a six-membered heteroaromatic ring. Compounds having MIC values 99% of cells were colonocytes. [3H]Thymidine uptake studies demonstrate a fivefold increased incorporation in this cell preparation compared to earlier fractions. 3-Isobutyl-l-methylxanthine (IBMX, 100 microM) was added to all cell suspensions in order to prevent cAMP metabolism. Cell suspensions were incubated for 2 min at 37 degreesC with different concentrations of 5-HT (n = 7). cAMP was measured by enzyme immunoassay. In another series of experiments, 5-HT (0.3 microM) stimulation of cAMP was similarly measured in the presence and absence of 5-HT receptor antagonists: 10 microM 5-HTP-DP (5-HT1P; n = 4), 0.1 microM ketanserin (5-HT2A; n = 4), 0.3 microM ondansetron (5-HT3; n = 4), 3 microM tropisetron (5-HT3 and 5 HT4; n = 4), and 10 nM GR-113808 (5-HT4; n = 5). Results. 5-HT produced a dose dependent increase in cAMP. The increase was significant at concentrations >/=0.3 microM when compared to cells incubated with IBMX alone. In the second series of experiment, 5-HT-induced generation of cAMP at a dose of 0.3 microM was significantly inhibited in the presence of GR-113808 and tropisetron. Conclusion. 5-HT acts at a 5-HT4 receptor to induce production of cAMP in rat distal crypt colonocytes. PMID- 9733601 TI - A paradox of cerebral hyperperfusion in the face of cerebral hypotension: the effect of perfusion pressure on cerebral blood flow and metabolism during normothermic cardiopulmonary bypass. AB - BACKGROUND: The purpose of this study was to determine the impact of perfusion pressure on cerebral blood flow (CBF) and metabolism during normothermic cardiopulmonary bypass (CPB) and after weaning. MATERIALS AND METHODS: Two groups of mongrel dogs were studied (Group A, CPB perfusion at 50 mm Hg, n = 6; and Group B, CPB perfusion at 100 mm Hg, n = 6). All animals underwent 2 h of normothermic bypass at cardiac indexes >2.1 L/min/m2 and were weaned from pump, maintained at pressures >75 mm Hg, and followed for an additional 2 h. RESULTS: In both groups CBF increased over 85% from baseline, in proportion to the hemodilution during the initiation of CPB. Intracranial pressure increased moderately in both groups during CPB, compromising CBF at 1 h in Group A, but not in Group B. The Group A cerebral metabolic rate for oxygen (CMRO2), however, remained unchanged as the percentage of oxygen extraction increased to compensate for the decreased CBF. During recovery, temperature, mean arterial pressure, and cerebral perfusion pressure were not significantly different between the two groups. However, the CBF, percentage of oxygen extracted, and CMRO2 were significantly lower in Group A. CONCLUSIONS: Normothermic CPB initiated with a crystalloid prime and performed at the lower end of a 50-70 mm Hg perfusion window resulted in a highly significant increase in CBF in order to compensate for hemodilution, while at the same time reduced the perfusion pressure available to supply the increased CBF. Together, these two events create a hemodynamic paradox of hyperperfusion in the face of hypotension. The reduction in CMRO2 in Group A is yet to be explained but seems to remain coupled to CBF and could represent a previously undescribed protective mechanism of hibernating cerebral tissue, similar to the phenomena of ischemic preconditioning in the heart, where cerebral tissue is stimulated to lower metabolism in response to inadequate CBF. PMID- 9733602 TI - Vasopressin selectively increases 5-fluorouracil uptake by colorectal liver metastases following hepatic artery bolus infusion. AB - BACKGROUND: Poor drug uptake secondary to the hypovascularity of colorectal liver metastases may partially explain their limited response to hepatic artery chemotherapy. Vasoconstrictors can increase tumor perfusion but their effect on drug uptake has not been well-characterized. The aim of this study was to determine whether vasopressin could selectively increase tumor uptake of 5-FU. MATERIALS AND METHODS: A syngeneic rat model of colorectal liver metastases was used. Control group rats underwent a 60-s hepatic artery infusion of 14C-5-FU (30 mCi/150 microL). Treatment group rats had vasopressin (60 mIU/kg, dose determined in pilot study) added to the 14C-5-FU infusion. Mean systemic arterial pressure was minimally affected. Tumor:liver (T/L UR) and tumor center:periphery (C/P UR) 5-FU uptake ratios were determined using quantitative autoradiography techniques. Differences in tumor size (< or > 4 mm) and location (superficial vs deep) were accounted for. Statistical analysis was by repeated measures ANOVA (P = 0.01 significant). RESULTS: A total of 161 tumors in 18 rats was analyzed. T/L URs were significantly higher in the treatment group compared to controls for tumors <4 mm (1.72 +/- 0.14 vs 0.70 +/- 0.16, P <0.001), tumors >4 mm (0.99 +/- 0.15 vs 0.45 +/- 0.16, P = 0.01), deep tumors (1.17 +/- 0.13 vs 0.68 +/- 0.15, P = 0.01), and superficial tumors (1.54 +/- 0. 15 vs 0.47 +/- 0.17, P <0.001). C/P URs did not differ significantly between the groups. CONCLUSIONS: The results of this study show that vasopressin selectively enhances the uptake of 5-FU by colorectal liver metastases in a rat model of hepatic artery infusion. This may represent a promising strategy for improving tumor response rates and patient survival. PMID- 9733603 TI - Interleukin-10 reduces morbidity and mortality in murine multiple organ dysfunction syndrome (MODS). AB - HYPOTHESIS: IL-10 will reduce morbidity and mortality in murine MODS. Introduction. Intraperitoneal (ip) zymosan causes a triphasic inflammatory process leading to MODS. Phase I is an acute systemic inflammatory response to sterile peritonitis. Phase II is the recovery phase. Phase III is characterized by recurrent illness, progressive organ dysfunction, and elevated proinflammatory cytokines. METHODS: Male ICR mice were randomized (on Experiment Day 0, time = 0 h) into four initial groups (A-D): Control Group A received no zymosan and no IL 10. Group B received zymosan (1 mg/g mouse BW, t = 0) and no IL-10. Group C received no zymosan and IL-10 at t = 2 h. Group D received zymosan and IL-10 at t = 2 h. On Experiment Day 4, mice in Groups B-D were randomized into six further treatment groups (B1 and B2, C1 and C2, D1 and D2). Group B1 received no treatment. Group B2 received IL-10 when clinical signs of recurrent illness developed (Phase III, 12-18 days after zymosan treatment). Mice were sacrificed when they were preterminal (clinical signs of shaking, shivering, or paralysis) or on Experiment Day 28 (survivors). Plasma total bilirubin and creatinine levels were measures of organ function. Terminal pulmonary compliance was measured in situ through a physiologic range of tidal volumes. RESULTS: Mice entering Phase III consistently progressed to MODS characterized by elevated bilirubin and hemorrhagic lungs which, if left untreated, was lethal. Mice treated with IL-10 (Group B2) when they entered Phase III had lower mortality (28.6% vs 100%, P < 0.02), longer survival (25 vs 18 days, P < 0.05), and improved lung pulmonary compliance (slope beta1 = 0.082 ml/mm Hg vs 0.059 ml/mm Hg, P < 0.001) compared to untreated (Group B1) mice in Phase III. CONCLUSIONS: IL-10 improves survival even when given after clinical signs of illness are present. PMID- 9733604 TI - Basic fibroblast growth factor expression precedes flow-induced arterial enlargement. AB - Arteries enlarge in response to increased blood flow, but the molecular signals controlling this enlargement are not well understood. Basic fibroblast growth factor (bFGF) is a potent mitogen for endothelial cells (EC) and smooth muscle cells (SMC) and promotes cellular proliferation and differentiation. In order to determine whether bFGF is expressed in response to increased blood flow in vivo, carotid-jugular arteriovenous fistulas (AVF) were created in Japanese white rabbits. The carotid artery proximal to the fistula was harvested after 1, 3, or 7 days and compared to nonoperated, control carotid arteries. Arterial blood flow increased five- to eightfold in all AVF animals and resulted in a significant increase in wall shear stress. The proximal carotid artery arterial diameter was no different from control after 1 and 3 days (2.3 +/- 0. 1 mm) but enlarged to 2.9 +/- 0.1 mm (P < 0.05) after 7 days. RT-PCR revealed early transcription of bFGF mRNA at 1 and 3 days with increased densitometric band ratio (bFGF/beta actin) at 7 days. Immunohistochemical analysis revealed bFGF protein localization in EC of control arteries as well as AVF arteries at all time points. SMC and adventitia expression of bFGF was absent in controls, minimal at 1 day, and increased after 3 and 7 days in the experimental groups. Western blotting confirmed the presence of bFGF in samples and transmission immunoelectron microscopy confirmed its nuclear localization. Endothelial cells in vivo express bFGF under both normal and elevated flow conditions. Smooth muscle cells, however, do not express bFGF under normal flow conditions but begin to express bFGF after 1 day of high flow with increased expression after 3 and 7 days. Flow induced arterial enlargement begins after SMC expression of bFGF. Therefore, bFGF may play a role in arterial enlargement and adaptive remodeling in response to increased flow. PMID- 9733605 TI - Curcumin blocks cyclosporine A-resistant CD28 costimulatory pathway of human T cell proliferation. AB - INTRODUCTION: Curcumin (Cur) is a phenolic component of common spice, turmeric. We have reported earlier that it possesses antineoplastic and immunosuppressive properties in vitro. It has been reported that cyclosporine A (CyA), a commonly used immunosuppressant does not inhibit CD28 costimulatory pathway of T-cell activation. We hypothesized that Cur, a tyrosine kinase inhibitor, would block CyA-resistant CD28 costimulatory pathway of human T cell proliferation. MATERIALS AND METHODS: Human T-lymphocytes were isolated from healthy donors using gradient centrifugation and rosetting techniques. In four separate experiments T-cells were plated in triplicate in 96-well plates at a density of 2X105 cells/well. These cells were stimulated with 0.5 ng/ml phorbol myristate acetate (PMA) + 0.5 (g/ml anti-CD28 antibody (PMA-CD28 group) or 2.5 microgram/ml PHA (PHA group). Cur or CyA at varying concentrations (0.31, 0.625, 1.25, 2.5, 5, or 10 microgram/ml and 1.25, 2.5, 5, 10, 20, or 250 ng/ml, respectively) was added and cellular proliferation was measured by the uptake of [3H]thymidine and is reported (mean cpm/well(SD). Cells from the PMA-CD28 group that were treated with either curcumin or 0.4% DMSO (vehicle control for curcumin) were studied for evidence of apoptosis by staining with viable dyes MC540 and Hoechst 33342 and subsequently analyzed in the cell sorter. RESULTS: Cur caused a concentration dependent inhibition of T-cell proliferation in the PMA-CD28 group (from 32775 +/ 3084 to 66 +/- 42 at 5.0 microgram/ml of cur) and PHA group (from 50956 +/- 5747 to 24 +/- 12 at 5.0 microgram/ml) with a calculated ED50 of 3.5 and 7.7, microM respectively. CyA inhibited T-cell proliferation in the PHA group with a calculated ED50 of 2.7 ng/ml but failed to block PMA + anti-CD28-stimulated T cell proliferation even at 250 ng/ml. PMA-CD28 group cells treated with 10 microgram/ml curcumin showed a significantly increased apoptosis as compared to control (0.4% DMSO). CONCLUSION: Since Cur blocks the CyA-resistant PMA + anti CD28 pathway of T-cell proliferation, it may have novel adjuvant immunosuppressive properties. PMID- 9733607 TI - Cell-free hemoglobin preserves renal function during normothermic ischemia. AB - INTRODUCTION: The purpose of this study is to determine whether an infusion of polymerized hemoglobin solution is capable of suppressing the tubular damage and loss of renal function normally seen during a clinically relevant period of warm ischemia. METHODS: Male rats (350-450 g) were randomized to treatment with control (5% human serum albumin, HSA, n = 6) or test solution (9% polymerized hemoglobin, PHB, n = 6). Following a right nephrectomy, the left renal artery was perfused with 4 ml of HSA or PHB at 37 degreesC. The left renal artery was temporarily occluded for 50 min. At 72 h, creatinine (Cr), blood urea nitrogen (BUN), and percentage hemoglobin (Hb) were measured and the kidney was removed. Stained kidney sections were graded for ischemic injury (0-4, 0 = normal and 4 = necrosis of the proximal tubule). All results were expressed as means +/- SEM and statistical analysis was performed by t test. RESULTS: Treatment with PHB resulted in lower Cr (1.2 +/- 0.23 mg/dl vs 3.26 +/- 0.60 mg/dl, P < 0.01) and BUN (60.5 +/- 12.7 mg/dl vs 151 +/- 20.2 mg/dl, P < 0.01) at 72 h compared to HSA controls. Total hemoglobin was not significantly different at 72 h. The weight of all treated kidneys increased; however, the increase was significantly less in the PHB-treated group (34 +/- 9.1% vs 70 +/- 7. 4%, P < 0.01). PHB-treated kidneys had less evidence of histologic damage compared to those in the HSA group (0.75 +/- 0.11 vs 2.50 +/- 0.64, P < 0.05). CONCLUSIONS: During normothermic renal ischemia, renal artery infusion of PHB resulted in preservation of renal function and histologic architecture. PHB solutions may be useful in preserving organ function during prolonged periods of in vivo ischemia. PMID- 9733606 TI - Flow cytometric analysis of chimerism in the rat tolerant to a renal allograft. AB - BACKGROUND: Chimerism, produced by the two-way migration of cells between graft and host, is a proposed mechanism by which tolerance occurs. The appearance of donor/recipient chimeras in tolerant ACI to Lewis rat heterotopic renal transplants was assessed in peripheral blood leukocytes using flow cytometry after staining with monoclonal antibodies. MATERIALS AND METHODS: ACI and Lewis rats were used as donor and recipient, respectively, after Rapamycin and Cyclosporin immunosuppression with or without donor blood or bone marrow transfusion. ACI and Lewis animals were also used for isograft and single-kidney controls. Animals were sacrificed at various time points after initial operation. Flow cytometry was performed on isolated peripheral blood leukocytes at sacrifice. Histologic and functional data were also obtained. The monoclonal antibody panel included RT1(a) (ACI, MHC I) combined with CD2, CD4, CD8, CD16, and CD25 or RT1(a,c) (bone marrow chimeras). RESULTS: RT1(a)+, CD8+ cells were transiently present in the peripheral blood leukocytes of Lewis recipients with the exception of allogeneic bone marrow recipients. No significant number of RT1(a)+, CD16+ ("dendritic" cell-line) chimeras was seen. Veto cells (RT1(a,c)+) were transiently present in the bone marrow recipients, but they did not lead to improved outcome. Furthermore, no correlation was made between histologic tolerance and any of these donor-derived cells. CONCLUSION: Donor/recipient chimerism, and the veto cell phenomenon are not operational tolerance mechanisms in this stringent model of ACI to Lewis rat renal transplantation. PMID- 9733609 TI - Choledochosphincter inhibitory reflex: identification of the reflex in dogs and its significance. AB - BACKGROUND: The sphincter of Oddi (SO) may undergo functional disorders. The mechanism of action of this sphincter is as yet not fully explored; the current study aims at studying some aspects of this mechanism. METHODS: Twelve mongrel dogs (mean weight 15.3 +/- 2. 9 SD kg, 8 male, 4 female) were studied. Under general anesthesia, the abdomen was opened and the gall bladder, common bile duct (CBD), and duodenum were exposed. Through separate punctures in the CBD, a balloon-tipped 2F catheter was introduced into the CBD and a 2F manometric catheter was placed within the SO. The positioning of the catheters was controlled fluoroscopically. The pressure response of the CBD and SO to CBD balloon distension with CO2, without and with separate anesthetization of either the CBD or SO, was recorded. RESULTS: Upon CBD distension by 0.5 ml of CO2, the pressure in the CBD rose (P < 0.001) and in the SO dropped (P < 0.01). The SO pressure drop was momentary and did not change significantly (P > 0. 05) with increase in the volume of CBD distension. The pressure response was blocked on separate anesthetization of the CBD and SO. CONCLUSIONS: The SO opening on CBD distension is suggested to be reflex and not hydromechanical. It seems to be mediated through a reflex which we call "choledochosphincter inhibitory reflex." Derangement of this reflex might result in functional disorders of the SO. PMID- 9733608 TI - Sodium butyrate upregulates Kupffer cell PGE2 production and modulates immune function. AB - The immunosuppressive effect of portal venous blood transfusions in organ transplantation has been well established and may be mediated by increased Kupffer cell production of the immunosuppressive arachidonic acid metabolite prostaglandin E2 (PGE2). In this study, butyrate, a short-chain fatty acid known to enhance gene transcription, is hypothesized to enhance Kupffer cell PGE2 production by altering cyclooxygenase or phospholipase A2 (PLA2) activity, thus augmenting the immunosuppressive effect of portal venous transfusion. Lewis rats were given a portal venous transfusion of Wistar-Firth blood or saline 1 h prior to Kupffer cell harvest. The in vitro effects of butyrate on Kupffer cell PGE2 production, cyclooxygenase, and PLA2 activity were assessed. Kupffer cell tumor necrosis factor-alpha (TNFalpha) production was also assessed due to its sensitivity to PGE2 and its proinflamatory effects. Kupffer cells from portally transfused animals produced significantly more PGE2 than saline-transfused controls. Addition of butyrate to the culture medium further increased PGE2 production by as much as sevenfold in Kupffer cells of portally transfused animals. Other short-chain fatty acids, propionate and hexanoate, did not increase PGE2 production. Butyrate added to Kupffer cells from transfused animals slightly upregulated inducible cyclooxygenase (COX-2) mRNA levels as measured by both Northern blot and reverse-transcriptase polymerase chain reaction and increased PLA2 activity fivefold as measured by Western blot. Kupffer cell immune function was also affected by in vitro butyrate treatment with a significant decrease in the production of TNFalpha. Thus, butyrate may be a useful immunoregulatory agent in organ transplantation protocols which seek to enhance transcription of immunosuppressive molecules. PMID- 9733610 TI - Intermittent hepatic pedicle clamping reduces liver and lung injury. AB - BACKGROUND: Temporary occlusion of the hepatic hilum is used to control hemorrhage during liver resection, but can result in widespread organ dysfunction. MATERIALS AND METHODS: Adult male Sprague-Dawley rats were subjected to either continuous (Group C) or intermittent (Group R) hepatic ischemia. The total ischemia time (60 min) was divided into four 15-min periods in Group R. Blood and lung tissue specimens were collected 2 h after the induction of ischemia (early phase), and 24 h after the termination of ischemia (late phase). Plasma lactate dehydrogenase (LDH), aspartate aminotransferase (AST), alanine aminotransferase (ALT), tumor necrosis factor-alpha (TNF-alpha), and interleukin 6 (IL-6) concentrations were measured. Histologic sections were studied using hematoxylin and eosin, as well as naphthol AS-D chloroacetate esterase techniques. RESULTS: In the early phase, LDH, ALT, TNF-alpha, and IL-6 concentrations were significantly higher in Group C than in Group R. Pulmonary septal thickening and polymorphonuclear leukocyte infiltration were less severe in Group R than in Group C. These differences were significant in the late phase. CONCLUSIONS: Intermittent hepatic pedicle clamping reduces the ischemia reperfusion injury not only to the liver but also to the lungs. This technique may improve the outcome in patients undergoing liver resection. PMID- 9733611 TI - Sepsis increases lung glutamine synthetase expression in the tumor-bearing host. AB - Acute stresses such as trauma or endotoxemia augment GLN demand and are associated with increased release of this amino acid from skeletal muscle and lung as well as increased expression of glutamine synthetase (GS, the principal enzyme of GLN synthesis) in these tissues. Muscle GLN release is also increased during chronic catabolic states which are associated with depletion of lean body mass, such as starvation or malignancy. We hypothesized that the expression of GS in response to an acute stress would be altered in tumor-bearing rats (TBR) experiencing severe cachexia and therefore a previously heightened GLN demand. Male Fischer 344 rats were implanted with methylcholanthrene-induced fibrosarcoma tumors or underwent sham operations and pair-feeding (sham) with TBR partners. When tumor burden reached approximately 15% of carcass weight, animals received injections of either Escherichia coli lipopolysaccharide (LPS, 1 mg/kg body wt) or saline vehicle. Rats were sacrificed 8 h after injection and lung and muscle tissue were analyzed for GS mRNA and protein via Northern and Western blot techniques, respectively. LPS injection caused an equivalent 4- to 6-fold increase in lung and muscle GS mRNA in both TBR and sham rats (P < 0.01). LPS did not produce a significant increase in GS protein level in muscle tissue of either group or in lung tissue of sham rats. In contrast, endotoxin did lead to a 3.5 fold increase in GS protein levels in lung tissue of TBRs (P < 0.05). This increase in lung GS protein may signify the importance of the lung in maintaining GLN homeostasis during chronic catabolic states where muscle mass is diminished. PMID- 9733612 TI - Stereotactic core biopsy reduces the reexcision rate and the cost of mammographically detected cancer. AB - The management of patients with mammographic abnormalities is rapidly shifting from needle-localized surgical biopsy (NLB) to stereotactic core biopsy (SCB). The precise role of SCB in the management of nonpalpable breast cancer remains to be defined. The purpose of this study was to compare SCB to NLB in the diagnosis of mammographically detected breast cancer in women who underwent breast conserving surgery. The records of all patients with nonpalpable breast cancer who underwent breast-conserving surgery from 1/1/95 to 6/1/97 were analyzed with respect to method of diagnosis, time interval from detection to diagnosis and breast-conserving surgery, volume of breast tissue excised, margin status and reexcision rate, number of surgical procedures, and total charges and costs per patient. During a 30-month period, 117 patients with nonpalpable breast cancer underwent breast-conserving surgery. The diagnosis was made by NLB in 69 patients and SCB in 48 patients. The time from detection to diagnosis and breast conserving surgery was 1.7 +/- 0.5 and 8.1 +/- 1.2 days for SCB patients and 6. 8 +/- 1.3 and 16.9 +/- 2.3 days for NLB patients (P < 0.01). The volume of breast tissue removed was 117.9 +/- 5.6 cm3 for SCB patients versus 75.2 +/- 2.9 cm3 for NLB patients (P < 0.01). Three SCB patients (6%) had positive margins, while 38 NLB patients (55%) had positive margins (P < 0.01). Only 1 SCB patient (2%) was reexcised, while 34 NLB patients (50%) were reexcised (P < 0.01). Eighty-nine percent of SCB patients had a single surgical procedure compared to 39% of NLB patients (P < 0.001). Patients who underwent SCB had reduced total charges and total costs per patient compared to NLB patients ($11,700 +/- $554 and $3537 +/- $167 per SCB patient versus $15,654 +/- $706 and $4853 +/- $198 per NLB patient, P < 0. 0001). Stereotactic core biopsy shortens the time from detection at mammography to diagnosis and breast-conserving therapy, permits appropriate discussion of treatment alternatives, reduces the positive margin rate and reexcision rate, and may represent a significant cost savings in the management of nonpalpable breast cancer. PMID- 9733613 TI - Short-term outcome of chronic immunosuppression on the development of breast lesions in premenopausal heart and lung transplant patients. AB - The risk of development of breast lesions in patients on chronic immunosuppression is unknown. In order to assess this risk, a retrospective review was performed of the records of 87 women between the ages of 12 and 47 years who received thoracic organ transplant from 1987 to 1996 at our institution. Inclusion criteria consisted of patients who were premenopausal, had no previous history of breast disease, and survived for at least 1 year posttransplantation. All patients were on a triple immunosuppressive regimen consisting of cyclosporine, steroids, and azathioprine. Mean follow-up was 4 +/- 1.2 years with a range of 1-6 years. During this period, 21 patients (24%) with a mean age of 38 +/- 10 years had screening or diagnostic mammography. The remainder of patients with a mean age of 24 +/- 9 years were followed clinically. Overall, 10 patients (11%) developed a total of 17 palpable, solid lesions at 33 to 72 months posttransplantation. Fifteen of these lesions were surgically excised. Five of the patients had multiple lesions. Pathological examination of the specimens revealed fibroadenoma in nine, fibrocystic disease in four, low grade phylloides tumor in one, and T-cell lymphoma in one case. None of the patients have developed primary breast cancer during follow-up. In conclusion, short-term immunosuppression does not increase the risk of the development of benign breast lesions in young women after thoracic organ transplantation, but rather the distribution of benign lesions is similar in an age-matched population. There were several cases of multiple fibroadenomas in the transplant population, but mammography revealed no malignant disease in this age group and does not need to be utilized in this population beyond what is considered standard for immunocompetent patients. The long-term effect ofimmunosuppressive therapy on the developmentof breast cancer in this group remains to be defined. PMID- 9733614 TI - Retroviral gene transfer: effects on endothelial cell phenotype. AB - BACKGROUND: Endothelial cells (EC) are an attractive target for somatic cell gene therapy, both for the treatment of cardiovascular disease and for the systemic delivery of recombinant gene products directly into the circulation. Recent evidence, however, suggests that viral transduction may induce unfavorable changes in EC phenotype. We examined the proliferative capacity and cell adhesion molecule (CAM) profile of EC after retroviral gene transfer (GT), employing a clinically relevant ex vivo GT protocol. METHODS: Human umbilical vein EC (HUVEC, N = 14 isolates) were exposed to supernatants containing the MFG.nlsLACZ vector, which codes for a nuclear localized beta-galactosidase. Control HUVEC were exposed to empty virus (CRIP) or no virus (NT). Efficiency of GT was quantitated by direct counting of beta-galactosidase-stained cells on a grid. Proliferation was quantitated by a 1-week assay of viable cell counts. Expression of EC activation molecules (Class II major histocompatibility antigen [MHC II], E selectin, intercellular adhesion molecule-1 [ICAM-1], and vascular cell adhesion molecule-1 [VCAM-1]) was examined using fluorescent cytometry (FACS) at rest and after cytokine stimulation. RESULTS: GT was reproducibly efficient (mean 57%, range 40-77%) using sequential viral exposures without selection. NT, CRIP, and LACZ-transduced HUVEC exhibited identical FACS profiles for E-selectin, ICAM-1, VCAM-1, and MHC II at rest, consistent with a nonactivated state. Upregulation of expression by cytokine was quantitatively similar for all groups. Growth rates were likewise not different between groups. CONCLUSIONS: Retroviral vectors may be employed to achieve high percentages of transduced EC for ex vivo GT without the use of selection. Transduced EC generated in this fashion are not activated, demonstrate an unaltered pattern of inducible CAM expression, and exhibit normal cell growth. The effects of GT on target cell phenotype are likely to be both vector and protocol specific and should be carefully assessed in each case prior to in vivo applications. PMID- 9733615 TI - Kupffer cell-mediated lymphocyte apoptosis: a PGE2-dependent mechanism of portal venous transfusion-induced immunosuppression? AB - BACKGROUND: Kupffer cells, after exposure to alloantigen via the portal vein, mediate an immunosuppressive effect involving enhanced production of PGE2. We hypothesize that up-regulation of Kupffer cell CoA-independent transacylase (CoA IT) by portal venous transfusion (PVT) is a possible mechanism of increased PGE2 production. Additionally, enhanced lymphocyte apoptosis, a process known to be macrophage dependent and facilitated by PGE2, is postulated as a possible mechanism of PVT-induced, Kupffer cell-mediated immunosuppression. METHODS: Lewis rat Kupffer cells were isolated after portal venous infusion with 1 ml of Wistar Firth blood (PVT) or saline (PV sal). Kupffer cell PGE2 production and CoA-IT activity was assessed. Lymphocyte apoptosis after exposure to PVT or PV sal treated Kupffer cells was also assessed by flow cytometry. RESULTS: PVT-treated Kupffer cells produced significantly more PGE2 and had increased CoA-IT activity when compared to PV sal-treated Kupffer cells. Treatment of Kupffer cells with a selective inhibitor of CoA-IT significantly decreased PVT-induced Kupffer cell PGE2 production. Increased lymphocyte apoptosis was observed after coculture with PVT-treated Kupffer cells compared to PV sal-treated cells. CONCLUSIONS: PVT increases Kupffer cell PGE2 production via increased CoA-IT activity and induces Kupffer cell-mediated lymphocyte apoptosis. Lymphocyte apoptosis facilitated by Kupffer cells within the hepatic sinusoid may be an important mechanism of PVT induced immunosuppression in organ transplantation. PMID- 9733616 TI - Pulmonary LPS-binding protein (LBP) upregulation following LPS-mediated injury. AB - BACKGROUND: The acute respiratory distress syndrome (ARDS) causes significant morbidity and mortality among trauma patients. Although multiple factors have been implicated, pulmonary injury in this population may be due to inflammatory mediators released in response to stimuli such as endotoxin (LPS). LBP plays an integral part in LPS-mediated release of inflammatory cytokines and increased local expression of LBP as the result of a primary injury may prime the lung to secondary LPS-mediated damage. MATERIALS AND METHODS: To determine the magnitude of pulmonary LBP upregulation following LPS injury we challenged rats with either intravenous (IV) or intratracheal (IT) LPS. Animals from each group were euthanized at 1, 2, 4, and 8 h postchallenge. Lung LBP and CD14 mRNA levels were assayed by Northern blot. Serum and bronchoalveolar lavage (BAL) fluid were assayed for inflammatory cytokines (TNF-alpha, MCP-1, IL-1beta, IL-6, and IL-10) by ELISA. RESULTS: LBP and CD14 mRNA levels were found to increase significantly in lung tissue after both IV and IT LPS with the IV LPS animals having a greater increase over 8 h. Serum TNF-alpha was significantly elevated in the IV LPS group whereas very low levels were detected in the BAL. Only BAL TNF-alpha was increased in the IT group at 8 h. CONCLUSION: Local pulmonary LBP and CD14 mRNA are both upregulated after either systemic or local LPS exposure. Such upregulation may render thelung more susceptible to local immune overactivation and injury during subsequent exposures to LPS. PMID- 9733617 TI - Protein kinase C activation during Ca2+-independent vascular smooth muscle contraction. AB - The cellular signaling mechanisms that modulate the sustained vascular smooth muscle contractions that occur in vasospasm are not known. We and others have hypothesized that a kinase cascade involving protein kinase C (PKC) modulates sustained vascular smooth muscle contraction. The purpose of this investigation was to develop a model in which the traditional contractile pathways involving myosin light chain phosphorylation are not activated and determine if the PKC pathway is activated under these conditions. The phosphorylation of caldesmon, myosin light chain (MLC20), and the specific PKC substrate, MARCKS (myristoylated, alanine-rich C-kinase substrate) was measured in bovine carotid arterial smoothmuscle (BCASM) stimulated with phorbol 12,13-dibutyrate (PDBu) under Ca2+-containing and Ca2+-free conditions. PDBu stimulation led to increases in caldesmon and MARCKS phosphorylation to the same degree in the presence or absence of Ca2+. PDBu stimulation but did not lead to increases in MLC20 phosphorylation over basal levels in Ca2+-free conditions. Immunoblot analysis of BCASM using PKC isoform-specific antibodies demonstrated the presence of one "Ca2+- dependent" PKC isoform: alpha, and two of the "Ca2+-independent" isoforms: epsilon and zeta. These data suggest that Ca2+-independent isoforms of PKC may play a role in the sustained phase of BCASM contractions through a kinase cascade that involves caldesmon and MARCKS phosphorylation but not MLC20 phosphorylation. PMID- 9733618 TI - Hepatocyte growth factor promotes liver regeneration with prompt improvement of hyperbilirubinemia in hepatectomized cholestatic rats. AB - BACKGROUND: In hepatectomy for patients with liver cirrhosis or cholestasis, prolonged postoperative hyperbilirubinemia is a troublesome complication and, if uncontrolled, often leads to life-threatening hepatic failure. Hepatocyte growth factor (HGF), first identified as the most potent mitogen for primary hepatocytes, has been shown to have multiple biological properties on liver, including mitogenic, antifibrotic, and cytoprotective activities. This study investigated the beneficial effects of a perioperative HGF supply to jaundiced liver after hepatectomy in rats. MATERIALS AND METHODS: As a model of jaundiced liver, we used an alpha-naphtylisocyocyanate (ANIT)-induced intrahepatic cholestasis model. Forty-eight hours after intraperitoneal injection of ANIT (75 mg/kg), when the total serum bilirubin level was moderately increased, a 70 % hepatectomy was performed. Human recombinant HGF (250 microgram/kg) (n = 15) or vehicle alone (n = 15) was intermittently administered to the rats 12 h before surgery and every 12 h after that until sacrifice. RESULTS: Perioperative HGF treatment effectively accelerated hepatocellular DNA synthesis of cholestatic liver followed by increase in the regenerated liver weight. Moreover, HGF supply promptly improved hyperbilirubinemia within 24 h after surgery. Histological examination revealed that HGF administration attenuated periportal inflammation and formation of bile duct obstructions. Postoperative serum concentrations of tumor necrosis factor-alpha, a representative inflammatory cytokine, were not altered by HGF treatment. CONCLUSIONS: Perioperative HGF supply not only promotes liver regeneration but also ameliorates hyperbilirubinemia in hepatectomized cholestatic rats. This mode of HGF treatment may be clinically useful for hepatectomy in patients with cholestasis. PMID- 9733619 TI - The effect of twist on microvascular anastomotic patency and angiographic luminal dimensions. AB - BACKGROUND: Microvascular anastomoses must be constructed perfectly in order to be successful. One of the subtle technical errors that can occur during construction is twisting of the anastomosis. In the present study, we examined the effect of twist on the immediate, 2-h postoperative angiographic dimensions and patency of microvascular anastomoses. MATERIALS AND METHODS: Sixty-four Sprague-Dawley rats were assigned randomly to four groups. The femoral arteries were dissected for a distance sufficient to permit the application of an 8.5-mm long microanastomotic approximator clamp. Microarteriorrhaphies were performed with twists of 0 degrees, 90 degrees, 180 degrees, or 270 degrees. Patency was assessed 2 h after surgery using transabdominal aortic arteriography with run off. Measurements were recorded for each anastomosis, as well as for the narrowest and widest diameters of the vessels within 10 mm of the anastomosis. RESULTS: Fifty-nine of the 64 rats had technically satisfactory angiograms which permitted measurement of vascular dimensions. The cross-sectional areas of the narrowest areas and the anastomoses were inversely related to the degree of twist, and were significantly reduced at 270 degrees (P < 0.05). Two-hour patency rates were 86% with 0 degrees twist, 82% with 90 degrees twist, 71% with 180 degrees twist, and 33% with 270 degrees twist. The reduction in patency with 270 degrees twist was statistically significant (P < 0.05). CONCLUSIONS: Twists of 0 degrees, 90 degrees, and 180 degrees did not impair patency at a statistically significant level, but twists of 270 degrees did significantly reduce patency of microvascular anastomoses (P < 0.05). PMID- 9733620 TI - Feasibility of double-expression retroviral vector using complement regulatory factor gene. AB - The donor source of vascular endothelial cells for hybrid blood vessels seeded with genetically engineered endothelial cells is generally considered to be autologous. The purpose of this study was to determine whether porcine endothelial cells transduced with double-expression retroviral vector using complement-resistant gene could be substituted for autologous endothelial cells. Decay-accelerating factor (DAF) and tissue plasminogen activator (tPA) cDNA were inserted into retroviral vector with homologous restriction factor 20 cDNA as a complement regulatory factor gene. Porcine aortic endothelial cells were transduced with these double-expression retroviral vectors, followed by the complement-dependent selection. Porcine endothelial cells transduced withdouble expression retroviral vectors showed a high gene expression of both DAF and tPA. Complement-dependent cytotoxicity and adherence of U937 were significantly inhibited by the transduction of double-expression vectors with complement regulatory factor gene. Double-expression retroviral vector using complement regulatory factor gene was efficacious in substituting porcine endothelial cells for the autologous endothelial cells. PMID- 9733621 TI - Osteogenic potential of cultured periosteal cells in a distracted bone gap in rabbits. AB - Osteogenesis of cultured periosteal cells was investigated in a bone-defective gap that was created artificially by distraction in the tibia of 12-week-old rabbits. A 10-mm circumferential length of periosteum was stripped from each stump of the osteotomized tibia, and the tibia was distracted rapidly (2 mm/day), resulting in disturbance of callus formation. Periosteal-derived cells, which were isolated from the contralateral tibia, were introduced into cell culture, subcultured twice to a population of 5 x 10(7) cells, and then injected into the defective bone gap when distraction was complete. Following inoculation of the cultured cells, significant new bone formation in the bone gap was observed. The control group which did not undergo cell transplantation showed only slight new callus formation which is supposed to be formed by osteogenic cells from the bone marrow. The bone mineral content of newly formed bone between the distracted tibia was analyzed quantitatively on radiographs. Histologically, the transplanted cells initially formed a mass at the injected site and then gradually differentiated into bone tissue from the peripheral region. Bromodeoxyuridine immunohistochemical stain was utilized to investigate the localization of the transplanted cells. The present study confirms that the orthotopically implanted periosteum-derived cells facilitate osteogenesis in a bone defect created using distraction in rabbits. PMID- 9733622 TI - Enhancement of metastatic activity of colon cancer as influenced by expression of cell surface antigens. AB - BACKGROUND: Cell surface antigens are contributory factors toward metastatic activity. There have been no detailed studies on changes in cell surface antigens of colon cancer cell lines. To control life-threatening metastasis, it is necessary to evaluate what types of changes in cell surface antigens exert an influence on metastatic activity. MATERIALS AND METHODS: In vivo selection was performed using the human colon cancer-derived cell line KM12SM to obtain variants of metastatic activity. A murine spleen injection-liver metastasis procedure reflecting the latter half of the metastatic process was adopted and repeated four times. Flow cytometric analyses were carried out to detect expression of antigens: Lewis a (Lea), Lewis x (Lex), sialyl Lewis a (sLea), sialyl Lewis x (sLex), E-cadherin, CD44v6, integrin alpha2 (CD49b), integrin alpha3 (CD49c), integrin alpha4 (CD49d), integrin alpha5 (CD49e), and integrin beta1 (CD29). RESULTS: In vivo selection produced variants with higher metastatic activity. In the original line KM12SM, sLea, E-cadherin, CD49b, CD49c, or CD29 were positive in more than 40% of the cells. After selection, the percentage of cells positive for Lea, sLea, and all examined integrins significantly increased. Lex, sLex, and CD44v6 increased slightly, while E-cadherin decreased slightly. CONCLUSIONS: In vivo selection and flow cytometric analysis revealed that Lea, sLea, CD49b, CD49c, and CD29 appear to be involved in the increase of metastatic activity. The changes of integrin expression in this study suggest that integrins collaborate in the promotion of adhesion to an extracellular matrix. PMID- 9733623 TI - In vivo gene transfer to skin and wound by microseeding. AB - BACKGROUND: Gene transfer to skin has many potential applications but lacks a safe, practical delivery method. This report presents a new technique, microseeding, for in vivo gene transfer to skin and wounds and for DNA-mediated vaccination. The plasmid DNA solution was delivered directly to the target cells of the skin by a set of oscillating solid microneedles driven by a modified tattooing device. MATERIALS AND METHODS: Skin and partial-thickness excisional wounds in pigs were microseeded with either hEGF expression plasmid or beta galactosidase expression plasmid. Human EGF was also delivered by single injection or particle bombardment. hEGF expression in wound fluid and in target tissue was determined by ELISA with anti-hEGF-specific antibodies. Additionally, weanling pigs were microseeded with a hemagglutinin of swine influenza virus expression plasmid and production of anti-HA-specific antibodies was determined by blocking ELISA. RESULTS: hEGF expression in microseeded partial thickness wounds (5664 pg/site) and skin sites (969 pg/site) peaked 2 days after transfection being four- to seven-fold higher than gene transfer by a single intradermal injection and two- to three-fold higher than particle-mediated gene transfer. The beta-galactosidase-expressing cells were detected in dermis and epidermis. Pigs microseeded with HA expression plasmid were protected from infection by the Swine influenza virus. CONCLUSIONS: These results demonstrate that microseeding is a simple and effective method for in vivo gene transfer to skin and wounds and is more efficient than single injection and particle-mediated gene transfer. PMID- 9733624 TI - Prevention of small intestinal ischemia-reperfusion injury in rat by anti cytokine-induced neutrophil chemoattractant monoclonal antibody. AB - The function of cytokine-induced neutrophil chemoattractant (CINC), which is the rat counterpart to human growth-related gene product belonging to the CXC chemokine subgroup, is based principally on neutrophil-specific chemotactic activity. In addition, we previously reported that plasma CINC was elevated during the period of small intestinal ischemia-reperfusion injury, and that there was a correlation between the degree of mucosal damage and the peak level of CINC after reperfusion, suggesting that CINC may play a major role in neutrophil infiltration into the rat small intestinal ischemia-reperfusion injury site. Thus, we investigated whether administration of anti-CINC monoclonal antibodies (mAbs) reduces small intestinal ischemia-reperfusion injury. Small intestine was subjected to ischemia for 3 h by occlusion of the anterior mesenteric artery with an atraumatic vascular clump. After infusion of anti-CINC mAbs or isotype-matched mAbs, the intestine was subjected to reperfusion. The pretreatment with anti-CINC mAbs attenuated ischemia-reperfusion injury in the small intestine, in association with the reduction of tumor necrosis factor-alpha and myeloperoxidase production, and resulted in the prolongation of survival. It is concluded that CINC plays an important role in the onset of rat small intestinal ischemia reperfusion injury. In addition, blocking the action of CINC, namely, the neutrophil chemotactic activity, may be useful in preventing ischemia-reperfusion injury in the small intestine. PMID- 9733625 TI - Thymic atrophy caused by thymocyte apoptosis in experimental severe acute pancreatitis. AB - Although a reduction in peripheral lymphocytes has been reported in clinical cases of acute pancreatitis, the thymic change remains still unknown. To investigate impairment of cellular immunity in acute pancreatitis, alterations of the thymus in rats with acute pancreatitis were examined experimentally. Male Wistar rats were used. Two groups with pancreatitis of different severity and a control group for each were established. The thymus was weighed and the number of thymocytes counted. Apoptosis in the thymus was examined by in situ nick-end labeling, DNA agarose gel electrophoresis, and cell cycle analysis using propidium iodide. Both thymus weight and number of thymocytes decreased significantly in the rats with necrotizing pancreatitis 20 h after induction of pancreatitis (P <0.02 vs sham operation). Neither thymus atrophy nor thymocyte reduction was observed in rats with edematous pancreatitis. In thymuses from rats with necrotizing pancreatitis, in situ nick-end labeling showed a significant increase in apoptotic changes of thymocytes, which was also confirmed by the stepladder pattern on agarose gel electrophoresis of the extracted DNA and by cell cycle analysis. It is concluded that thymus atrophy associated with apoptosis occurs in rats with necrotizing pancreatitis. PMID- 9733626 TI - The immunologic role of IFN-gamma in ACI to Lewis kidney transplantation. AB - BACKGROUND: One of the proposed mechanisms of tolerance induction is the Th-1/Th 2 paradigm. The Th-1 cell is proinflammatory, secreting IFN-gamma and IL-2. Conversely, the Th-2 cell is anti-inflammatory, secreting IL-4 and IL-10. In our earlier studies a shift toward Th-2 dominance was required for tolerance induction in this model. MATERIALS AND METHODS: ACI and Lewis rats were used as donors and recipients, respectively. Twelve hours prior to engraftment, rapamycin 1.5 mg/kg po and cyclosporin 10 mg/kg sc were given, followed by 5 mg/kg sc postop (days 1-7). Lewis rats were used as isografts. Functional allograft tolerance was induced consistently in 100% of the recipients with 50% of the allografts exhibiting normal histology beyond 120 days. Qualitative RT-PCR was performed on the grafts to determine IFN-gamma expression with beta-actin housekeeping gene as control. RESULTS: IFN-gamma was expressed in all untreated allografts (5/5) and all treated, yet rejecting, allografts (4/4). None of the isografts (0/5) or histologically tolerant allografts (0/4) expressed IFN-gamma. This distribution was statistically significant (P < 0.001, Fischer's exact test). CONCLUSION: Our findings support a shift from Th-2 to Th-1 predominance as the corollary mechanism responsible for preventing histologic tolerance. PMID- 9733627 TI - Tumor necrosis factor binding protein improves incisional wound healing in sepsis. AB - BACKGROUND: Sepsis is associated with poor wound healing; however, the exact role of tumor necrosis factor (TNF) as a mediator of sepsis-induced alterations in different types of tissue repair is unknown. This study examines the effects of a specific TNF antagonist (TNFbp) on the healing of intestinal anastomoses, incisional wounds, and polyvinyl (PVA) sponge implants in chronic abdominal sepsis. METHODS: Three groups of male Sprague-Dawley rats were studied: control, sepsis, and sepsis + TNFbp. Jejunal resection and anastomosis were performed through a 4-cm upper midline incision on day 1. On day 3, sepsis was induced by creation of a chronic abdominal abscess. Saline (0.1 ml) or TNFbp (1.0 mg/kg, 0.1 ml) was injected subcutaneously every day starting 4 h prior to sepsis. On day 7, the wound-breaking strength (WBS) of the skin incision and intestinal anastomoses was determined using a tensiometer. Wound histology and collagen deposition were evaluated by comparison of Sirius red-stained sections. The hydroxyproline content of PVA sponges was used to quantitate collagen content under the different experimental conditions. RESULTS: Septic mortality (20% vs 26%) was not significantly altered by TNFbp. Septic animals demonstrated a reduction in food consumption on days 3 to 5 that was not affected by TNFbp administration. Neither sepsis nor TNFbp altered the breaking strength or histologic appearance of intestinal anastomoses. However, the breaking strength of incisional wounds was decreased by 40% in septic rats (P < 0.001 vs controls). Administration of TNFbp to septic rats significantly improved incisional WBS (P < 0.01 vs sepsis), but not to control levels. Serius red staining of incisional wounds and PVA sponges demonstrated a decrease in collagen organization and deposition in septic rats that was ameliorated by TNFbp. Similarly, the reduction in hydroxyproline content of PVA sponges from septic animals was prevented by TNFbp. CONCLUSIONS: The process of tissue repair in intestine and skin wounds appears to be significantly different following the septic insult. The healing of jejunal anastomoses was refractory to the catabolic effects of sepsis. In contrast, collagen deposition and organization are significantly decreased in cutaneous wounds during chronic sepsis. TNFbp significantly ameliorated the inhibitory effects of sepsis on cutaneous wound healing. These results suggest that TNF is an important mediator of the decrease in collagen deposition observed in cutaneous wounds during the septic state. PMID- 9733628 TI - Prevention of retrosternal adhesion formation in a rabbit model using bioresorbable films of polyethylene glycol and polylactic acid. AB - The purpose of this study was to test the efficacy of three bioresorbable films of polyethylene glycol (EO) and polylactic acid (LA) (EO/LA = 1.5, 2.5, and 3.0) in the prevention of adhesion formation between the epicardium and the sternum (retrosternal adhesions) in a rabbit model. Retrosternal adhesions were generated by sternotomy, pericardiotomy, and abrasion of the anterior epicardium. The adhesion barrier was placed between the epicardium and the sternum and sutured to the edge of the pericardium. Epicardial adhesions were evaluated 14-20 days later by assessing the area of the epicardium covered by adhesions. In the control rabbits, tenacious adhesions were observed between sternum and the central portion of epicardium (portion exposed through the pericardiotomy) which were difficult to dissect. When a bioresorbable film was placed over the pericardium, adhesion formation at the central strip of the epicardium (area between the sternum and the epicardium exposed through the pericardium) could be reduced or prevented. At this site, the areas of adhesion formation were 0% (EO/LA = 1.5), 8.4 +/- 2.8% (EO/LA = 2.5), and 5.6 +/- 4.7% (EO/LA = 3.0) of the central strip, significantly less than that observed in the control group, 78.0 +/- 5.8% (P < 0.01). At the anterior left and right and posterior apex of the heart (sites where the film was not placed), there were no differences between control and treatment groups. The films were completely resorbed at the time of necropsy in group EO/LA = 2.5 and 3.0. Small pieces of film were observed in group EO/LA = 1.5. In conclusion, the bioresorbable films [EO/LA = 1.5 (REPEL-CV), 2.5, or 3.0] were efficacious in the reduction of retrosternal adhesions to the epicardium. PMID- 9733629 TI - Induction of apoptosis in a neuroblastoma and hepatocyte coculture model. AB - BACKGROUND: The dysregulation of apoptosis may alter the progression of tumor growth and explain the clinical dichotomy observed in children with neuroblastoma (NB). An overexpression of the bcl-2 proto-oncogene induces resistance to apoptosis and has been observed in unfavorable NB. We hypothesized that alterations in apoptosis may be a result of the interactions between NB and the tissues surrounding it. MATERIALS AND METHODS: Human Chang liver cells (HCL, 10(4) cells/cm2) were plated in two-chamber slides for 3 days. Human NB cells (10(5) cells/cm2) were added to one of the chambers and incubated for 3 more days. Control NB were plated under identical conditions in its own medium and in the HCL medium with growth curves measured. DNA fragmentation was detected via the TUNEL method (TdT-mediated nick end-labeling) and bcl-2 expression was determined by immunostaining. RESULTS: NB growth was unaltered by the change in medium. NB stained mildly positive for bcl-2 when plated alone but became markedly positive in coculture. Histologically, HCL and NB appeared healthy when plated alone, but a halo of apoptotic HCL was seen around NB in the coculture. When plated alone, both NB and HCL demonstrated minimal apoptotic activity as detected via the TUNEL method. In the coculture, a halo of HCL surrounding the NB exhibited markedly increased DNA fragmentation and this intensity diminished in cells distant from the NB. CONCLUSIONS: The regulation of apoptosis was altered in this coculture model of NB and HCL. HCL stimulated NB to overexpress bcl-2 and presumably become resistant to apoptosis. Conversely, NB induced the surrounding HCL to undergo apoptosis. The interaction between the local tissue and NB induced alterations in apoptosis in both cell types and resulted in a survival advantage for NB. PMID- 9733630 TI - Lecithin protects against plasma membrane disruption by bile salts. AB - INTRODUCTION: Detergent disruption of epithelial plasma membranes by bile salts may contribute to pathogenesis of cholestasis and gastroesophageal reflux disease. Bile, despite containing high concentrations of bile salts, normally is not toxic to biliary or intestinal epithelia. We hypothesize that lecithin in bile may protect cell membranes from disruption by bile salts. METHODS: We studied the interactions of taurine conjugates of ursodeoxycholate (TUDCA), cholate (TCA), chenodeoxycholate (TCDCA), and deoxycholate (TDCA) with erythrocyte plasma membranes with or without large unilamellar egg lecithin vesicles for various times at 23 degreesC. Release of hemoglobin was quantified spectrophotometrically. The concentration of bile salt monomers and simple micelles in the intermixed micellar aqueous phase (IMMC) was determined by centrifugal ultrafiltration. RESULTS: The degree of hemolysis depended on the hydrophobicity of the bile salts and was progressive over time. Addition of lecithin reduced the hemolytic effects of 20 mM TCA or 2 mM TDCA in a concentration-dependent manner at both 30 min and 4 h. Increasing the concentration of lecithin progressively reduced the IMMC of TDCA. Hemolysis following addition of lecithin to 2 mM TDCA was comparable to hemolysis produced by lecithin-free TDCA solutions when diluted to similar IMMC values. CONCLUSION: We conclude that lecithin reduces plasma membrane disruption by hydrophobic bile salts. This protection may be attributable to association of bile salts with vesicles and mixed micelles, reducing the concentration of bile salt monomers and simple micelles available to interact with cell membranes. Lecithin may play a key role in preventing bile salt injury of biliary and gastrointestinal epithelia. PMID- 9733631 TI - Nitric oxide mediates acute lung injury by modulation of inflammation. AB - Nitric Oxide's (NO) function in vasomotor control, inflammation, and signal transduction makes it an attractive potential mediator of the capillary leak seen in acute lung injury. Despite extensive study, the role of NO in intestinal ischemia/reperfusion-induced capillary leak remains controversial. Rats were treated with vehicle, norepinephrine, or L-NNA (nitric oxide synthase inhibitor) and then underwent sham laparotomy or 30 min SMA occlusion followed by 1 to 12 h of reperfusion. Evan's Blue dye was administered 1 h before animals were euthanized. Ratios of bronchoalveolar lavage or small-intestine lavage to serum dye concentrations were calculated as measures of capillary leak. Circulating neutrophil activation was measured with a nitroblue tetrazolium reduction assay. In vehicle-treated animals, both capillary leakage and PMN activation peaked at 4 h of reperfusion. These parameters returned to baseline by 12 h. Treatment with L NNA accelerated ischemia/reperfusion-induced PMN activation as well as accelerated capillary leak from 4 to 1 h. Treatment with norepinephrine (hypertensive control) increased the magnitude of lung capillary leak but had no effect on the timing of ischemia/reperfusion-induced PMN activation or ischemia/reperfusion-induced capillary leak. These data show that intestinal ischemia/reperfusion-induced systemic capillary leak is associated with systemic neutrophil activation. Nitric oxide synthase inhibition accelerates ischemia/reperfusion-induced capillary leak and mediates the capillary leak seen in acute lung injury by modulating neutrophil activation. PMID- 9733632 TI - The effects of G-CSF treatment and starvation on bacterial translocation in hemorrhagic shock. AB - BACKGROUND: Bacterial translocation is thought to be responsible for infectious complications after hemorrhagic shock. The aim of this study is to investigate the effects of granulocyte colony-stimulating factor (G-CSF) treatment on bacterial translocation in starved or fed animals subjected to hemorrhagic shock. MATERIALS AND METHODS: Fifty Wistar albino rats (200-275 g) were divided into six groups such as naive control (n = 7), G-CSF treatment (n = 7), hemorrhagic shock in starved rats (n = 9), hemorrhagic shock in fed rats (n = 9), G-CSF treatment 24 h before hemorrhagic shock in starved rats (n = 9), and G-CSF treatment 20 min after hemorrhagic shock in fed rats (n = 9). Hemorrhagic shock was induced by withdrawal of 2.1 ml/100 g blood via a carotid arterial cannulae placed under sodium pentobarbital anesthesia. Twenty-four hours later, mesenteric lymph nodes, liver, spleen, and peripheral blood samples were evaluated by using a quantitative microbiological technique and the numbers of colony-forming units were compared between groups. RESULTS: No bacteria was detected in samples from naive controls or G-CSF-treated unshocked rats. In animals subjected to hemorrhage, Escherichia coli was the predominant pathogen together with Streptococcus faecalis, Pseudomonas, and Lactobacillus species. In this model, starvation augmented the magnitude of bacterial translocation while G-CSF treatment has virtually abolished it. CONCLUSION: Under experimental conditions, preshock starvation increases gut-derived bacterial translocation and administration of G-CSF before or after hemorrhagic insult significantly reduces it. PMID- 9733633 TI - The effect of sequential injuries on splanchnic perfusion and eicosanoid release. AB - BACKGROUND: This study examines the hypothesis that sequential burn injury followed by intraabdominal sepsis induces significantly greater splanchnic hypoperfusion and reduced intestinal PGI2 release than either injury independently. MATERIALS AND METHODS: Anesthetized Sprague-Dawley rats were randomized to one of four groups: BURN (45% body surface area scald burn) + cecal ligation and puncture (CLP); BURN alone; CLP alone; or uninjured controls (SHAM). Twenty-four hours following injury, superior mesenteric artery (SMA) blood flow was measured with a doppler flow probe. Splanchnic eicosanoid release (6-keto PGF1alpha, metabolite of PGE2; TxB2, metabolite of TxA2; and PGE2) was measured in mesenteric venous effluent utilizing an isolated, perfused bowel preparation. RESULTS: SMA blood flow was no different than that of controls 72 h following BURN injury alone; whereas CLP alone resulted in a 80% reduction in splanchnic blood flow when compared with controls (P < 0.001). SMA blood flow in animals sustaining BURN + CLP was only modestly reduced from controls (P = 0.04) and 3.6 times greater than that of animals sustaining CLP alone (P < 0.001). PGI2 was the dominant eicosanoid released by the intestine with levels 10 times greater than TxB2 and nearly 50 times greater than PGE2. CLP either alone or when combined with BURN was associated with a 60% decrease in splanchnic PGI2 release when compared to controls (P < 0.05). CONCLUSIONS: These data suggest that moderate BURN injury in rats attenuates the severe reduction in splanchnic perfusion associated with intraabdominal sepsis and that this occurs despite profound reductions in the release of the endogenous splanchnic vasodilator PGI2. PMID- 9733634 TI - Effect of lung volume reduction surgery on pulmonary diffusion capacity in a rabbit model of emphysema. AB - BACKGROUND: While there is renewed interest in lung volume reduction surgery (LVRS) for treatment of emphysema, many aspects of the operation such as patient selection and surgical end points of excision are uncertain. We studied the effects of LVRS on measured lung volumes and diffusion capacity in an animal model to investigate optimal resection volumes. METHODS: Emphysema was induced in 32 New Zealand white (NZW) rabbits using aerosolized elastase. Helium dilution lung volumes and single breath DLCO were measured concurrently at baseline, following induction of emphysema (preop), and 1 week postoperatively (postop) following LVRS. Bilateral upper and middle lobe stapled lung resections were performed through midline sternotomies with excision of variable amounts of lung tissue from 1.8 to 5.8 g. RESULTS: FRC increased following induction of emphysema and decreased postoperatively. DLCO improved with increasing lung tissue resection up to 3 g of tissue and then decreased as even greater amounts were removed (r = 0.54). CONCLUSIONS: Measured lung volumes increase with development of emphysema and appropriately decrease in response to LVRS in this rabbit model. DLCO improves with moderate resection but then decreases with excessive excision of lung quantities and may help define one physiologic operative end point. In this rabbit model, excision of approximately 30% of lung volume was optimal and prevented further decrease in diffusion capacity. PMID- 9733635 TI - Hepatocyte growth factor stimulates fetal gastric epithelial cell growth in vitro. AB - BACKGROUND: The growth and development of the fetal gastrointestinal tract is likely mediated, in part, by peptide growth factors. We compared the mitogenic effects of graded doses of hepatocyte growth factor (HGF) to epidermal growth factor (EGF), transforming growth factor-alpha (TGF-alpha), and insulin-like growth factor-1 (IGF-1) on fetal rabbit gastric epithelial cells. MATERIALS AND METHODS: Fetal rabbit gastric epithelial cells were purified by mechanical dissociation and selected culture and grown in short-term (24 h) and long-term (12 days) culture. Stimulation of fetal gastric epithelial cell growth in response to individual peptide growth factors was measured by [3H]thymidine incorporation and cell counting. RESULTS: In short-term culture, HGF stimulated [3H]thymidine incorporation in a dose-dependent manner from a threshold at 10 pM to a maximum at 100 pM. For EGF and TGF-alpha, maximal stimulation occurred at 100 pM. For HGF, maximal [3H]thymidine incorporation was 3.6 +/- 0.7 times basal. For EGF and TGF-alpha, maximal [3H]thymidine incorporation was 4.3 +/- 0.4, and 3.6 +/- 0.4 times basal, respectively. For IGF-1, maximal [3H]thymidine incorporation was only 70% of the maximal effect observed for the other growth factors tested. Rabbit amniotic fluid increased [3H]thymidine uptake in a dose dependent manner. In long-term culture, purification to greater than 90% epithelial cells was attained after 12 days treatment. For HGF, EGF, TGF-alpha, and 20% rabbit amniotic fluid, significant increases in cell number above control (P < 0.05) were observed at 1 nM concentrations. None of these individual factors, however, increased cell growth as significantly as that of 10% fetal bovine serum. CONCLUSIONS: Our results suggest that: (1) HGF stimulates [3H]thymidine uptake and cell proliferation in fetal rabbit gastric epithelial cells in vitro, and (2) HGF's mitogenic effect on fetal rabbit gastric epithelial cell growth is comparable to that observed for EGF and TGF-alpha, but superior to the effect observed for IGF-1. PMID- 9733636 TI - Assessment of the role of neutrophils on the antitumor effect of TNFalpha in an in vivo isolated limb perfusion model in sarcoma-bearing brown Norway rats. AB - INTRODUCTION: Isolated limb perfusion (ILP) with TNFalpha in combination with melphalan and IFNgamma has resulted in an immediate and dramatic tumor response in patients. Such an effect was also noted following ILP in a rat sarcoma model. This model enables us to investigate several factors responsible for the TNFalpha induced tumor responses. We applied total body irradiation (TBI) to reduce white blood cell count, to investigate the contribution of leukocytes to the anti-tumor effect of TNFalpha. METHODS: Small fragments of the nonimmunogenic BN 175 sarcoma were implanted sc in the lower hind leg. A 5 Gy TBI was performed before ILP at a tumor diameter of approximately 15 mm. The hind limbs of 63 rats were perfused and were divided into 6 groups: group 1, sham perfusion, n = 9; group 2, TBI + sham perfusion, n = 6; group 3, TNFalpha 50 microgram, n = 9; group 4, melphalan 40 microgram, n = 9; group 5, TNFalpha 50 microgram + melphalan 40 microgram, n = 22; group 6, TBI + TNFalpha + melphalan ILP, n = 8. In addition, 10 rats were perfused for histological analysis at 24 h post-ILP. RESULTS: We observed in Group 1: 9/9 progressive disease (PD); Group 2: 6/6 PD; Group 3: 9/9 PD; Group 4: 9/9 no change (NC) of tumor diameter for at least 4 days; Group 5: 6/22 NC, 16/22 complete remission (CR), 12/16 of which showed skin necrosis at the tumor site; and Group 6: 7/8 NC and 1/8 CR (without skin necrosis). After TBI, WBC reduction of 80-95% was observed, while the number of platelets was not significantly reduced and platelet aggregation was maintained at 72 %. Histological analysis revealed decreased hemorrhagic necrosis associated with the absence of PMN infiltration at the tumor margins in the TBI rats. CONCLUSION: TBI and the associated reduction in WBC count decreased the tumor response by TNFalpha and melphalan significantly and abrogated the immediate response of skin necrosis at the tumor site, as found in rats treated with TNFalpha and melphalan without TBI. These data strongly suggest that leukocytes play an important role in the hemorrhagic effects of TNFalpha. PMID- 9733637 TI - Crystal structure of agkistrodotoxin, a phospholipase A2-type presynaptic neurotoxin from agkistrodon halys pallas. AB - The crystal structure of agkistrodotoxin containing eight copies of molecules in the asymmetric unit has been determined at 2.8 A resolution to a crystallographic R factor of 0.207 by the molecular replacement technique. Two spatially adjacent regions of agkistrodotoxin molecule, turn 55-61 and stretch 85-91, are remarkably different from those of non-neurotoxic isoforms in conformation and electrostatic characteristics. These regions are likely to be involved in the recognition of agkistrodotoxin towards the specific receptor at the presynaptic membrane. The structural comparison of the interfacial recognition site with non-neurotoxic isoforms reveals a decreased hydrophobicity and lack of residues with bulky hydrophobic side-chains (i.e. Trp) to serve as membrane anchors. This structural feature of agkistrodotoxin may be related to the reduced non-specific binding of the toxin to non-targeted membrane before it arrives at the presynaptic membrane and recognizes the putative receptor. A unique hydrophobic patch including residues I19, P20, F21, A23, F24, M118 and F119 is found on the surface of the molecule near the entrance of the hydrophobic channel which plays an important role in crystal packing. The interaction mode between the patches might give a clue to the binding of the neurotoxin on the membrane. The agkistrodotoxin molecules in the asymmetric unit form two tetramers and each tetramer exhibits a novel "dimer of dimers"-like structure. A molecule-spanning four-stranded antiparallel beta-sheet is formed by the beta-wings of two molecules within a tetramer. PMID- 9733638 TI - Missense translation errors in Saccharomyces cerevisiae. AB - We describe the development of a novel plasmid-based assay for measuring the in vivo frequency of misincorporation of amino acids into polypeptide chains in the yeast Saccharomyces cerevisiae. The assay is based upon the measurement of the catalytic activity of an active site mutant of type III chloramphenicol acetyl transferase (CATIII) expressed in S. cerevisiae. A His195(CAC)-->Tyr195(UAC) mutant of CATIII is completely inactive, but catalytic activity can be restored by misincorporation of histidine at the mutant UAC codon. The average error frequency of misincorporation of histidine at this tyrosine UAC codon in wild type yeast strains was measured as 0. 5x10(-5) and this frequency was increased some 50-fold by growth in the presence of paromomycin, a known translational error-inducing antibiotic. A detectable frequency of misincorporation of histidine at a mutant Ala195 GCU codon was also measured as 2x10(-5), but in contrast to the Tyr195-->His195 misincorporation event, the frequency of histidine misincorporation at Ala195 GCU was not increased by paromomycin, inferring that this error did not result from miscognate codon-anticodon interaction. The His195 to Tyr195 missense error assay was used to demonstrate increased frequencies of missense error at codon 195 in SUP44 and SUP46 mutants. These two mutants have previously been shown to exhibit a translation termination error phenotype and the sup44+ and sup46+ genes encode the yeast ribosomal proteins S4 and S9, respectively. These data represent the first accurate in vivo measurement of a specific mistranslation event in a eukaryotic cell and directly confirm that the eukaryotic ribosome plays an important role in controlling missense errors arising from non-cognate codon-anticodon interactions. PMID- 9733639 TI - Roles of pIII in filamentous phage assembly. AB - Filamentous phage protein III (pIII), located at one end of the phage, is required for infectivity and stability of the particle. Cells infected with phage from which gene III has been completely deleted produce particles that are not released into the medium but stay associated at the surface. These particles are much longer than normal phage. They can be released by subsequent expression of pIII. Viewed with the electron microscope, cells infected with gene III deletion phage are decorated with structures that resemble extremely long pili. Surprisingly, such cells are viable and can form colonies. The pIII deficiency can be complemented in trans, but there is a threshold concentration below which assembly does not occur. Above this threshold, pIII is used very efficiently and is incorporated into infectious but longer than unit length phage. As the concentration of pIII is increased, the number of infectious particles increases, and their average length decreases.pIII stabilizes pVI, a second phage protein found at the pIII end of the particle. In the absence of pIII, degradation of pVI is very rapid. pIII is thus not only required for infectivity and particle stability, but to terminate assembly and release the phage from its assembly site. PMID- 9733640 TI - Manganese water-soluble porphyrin senses DNA conformation. AB - We have determined the sites that are preferentially cleaved by Mn(T4MPyP) (where T4MPyP is the dianion of 5, 10, 15, 20, tetrakis (4-N-methylpyridine)porphyrin) on synthetic DNAs and on both intrinsically curved and average-shaped natural DNA sequences. On the basis of cleavage selectivity and of DNase I footprinting we show that the recognition specificity by this compound is based on steric properties: the preferred conformation is a DNA minor groove narrower than average and dimensionally defined. This conclusion is reached on the basis of: (i) the localization of the preferential cleavage sites at the 3' extremity of short A-tracts, known to undergo minor groove directional narrowing; (ii) the effects of temperature on cleavage specificity on curved sequences; (iii) the localization of cleavage sites in synthetic constructs whose crystal and solution structure was previously defined, and in programmed sequence variants thereoff; (iv) the effects of base substitutions on cleavage efficiency; (v) DNase I footprinting analysis. Several of these evidences argue against the possibility that Mn(T4MPyP)/DNA site selection occurs on the basis of electrostatic potential effects.Mn(T4MPyP) provides a tool for the analysis of DNA conformation whose selectivity is complementary to that of DNase I and hydroxyl radicals. PMID- 9733641 TI - Sequence dependence of branch migratory minima. AB - The Holliday junction is a central intermediate in the process of genetic recombination. The position of its branch-point can relocate through an isomerization known as branch migration. This migration occurs because the branch point is flanked by homologous symmetry. All attempts at modeling the kinetics of branch migration have relied on the assumption that branch migration minima are sequence-independent. We have tested that assumption here, using a competition assay based on symmetric immobile branched junctions; these are junctions that cannot undergo branch migration, despite the fact that they are flanked by homology. The assay used is predicated on the non-association of strands displaced in the assay; we have tested this assumption, and have performed our experiments under conditions where we know that it is true. We have measured the free energy of relocating a branched junction from a fixed non-homologous sequence to all possible dimeric symmetric sequences. We find that the assumption of sequence-independence is often valid, but that it is not universally true. We find that the flanking sequences can have a marked effect on the free energy measured, both for extensions of symmetry and for reversals of flanking nucleotides. We have varied the temperature in our experiments, and have derived both enthalpies and entropies for the different sequences. The entropies are largely unfavorable, whereas the enthalpies are largely favorable; regardless of the signs of these quantities, we see that this is another system where enthalpy entropy compensation is operative. PMID- 9733642 TI - Large-scale sequence comparisons reveal unusually high levels of variation in the HLA-DQB1 locus in the class II region of the human MHC. AB - Comparison of genomic sequences flanking the HLA-DQB1 locus in the human MHC class II region reveals local sequence variation of up to 10%, which is the highest level of sequence variation found in the human genome so far. The variation is haplotype-specific and extends far beyond the transcriptional unit of the DQB1 gene, suggesting hitch-hiking along with functionally selected alleles as the most likely mechanism. All major insertions/deletions (indels) were found to be of retroviral origin and in the immediate upstream region of DQB1. Possible cis-acting effects of these indels on the transcriptional regulation of DQB1 are discussed. PMID- 9733643 TI - Nidogen-2: a new basement membrane protein with diverse binding properties. AB - Human nidogen-2 was cloned and sequenced (1375 residues) and found to share 46% sequence identity and a similar domain arrangement with the previously characterized basement membrane protein nidogen-1. Recombinant nidogen-2 was purified as a 200 kDa protein from transfected mammalian cell medium, showed a high level of N and O-glycosylation, and could be clearly distinguished from nidogen-1 (150 kDa) by specific antibodies. Electron microscopy demonstrated that the two isoforms have a similar shape, consisting of three globular domains connected by two threads, but differ somewhat in length. Northern blots and immunological assays demonstrated co-expression of the nidogens in various tissues and cultured cells. Immunofluoresence revealed colocalization in vessel walls and other basement membrane zones but some differences in heart and skeletal muscle. Nidogen-2 interacted with collagens I and IV, and perlecan at a comparable level to nidogen-1 but failed to bind to fibulins. Nidogen-2 bound to laminin-1, but only moderately to the epitope on the laminin gamma1 chain, which promotes high-affinity binding of nidogen-1. Both nidogens were cell-adhesive for a restricted number of cell lines, with nidogen-2 having a higher activity. Together, these data suggest that nidogen-2 can compensate for some but not all functional activities ascribed to nidogen-1. PMID- 9733644 TI - Characterization of nebulette and nebulin and emerging concepts of their roles for vertebrate Z-discs. AB - Nebulin is an 800 kDa large actin-binding protein specific to skeletal muscle and thought to act as a molecular template that regulates the length of thin filaments. Recently, a 100 kDa nebulin-like protein has been described in the avian cardiac muscle and referred to as nebulette. We have determined the full length (8 kb) cDNA sequence of the human nebulette. Its open reading frame (3044 bp) encodes a 109 kDa protein that shares extensive similarity with the C terminal region of human nebulin. The C-terminal regions of nebulin and nebulette are identical in domain organization and share a family of highly related C terminal repeats, a serine-rich domain with potential phosphorylation sites, and an SH3 domain. Immunoelectron-microscopy suggests that the C-terminal 30 kDa of nebulin and nebulette filaments integrate into the Z-disc lattice, whereas their N termini appear to project into the I-band. Gene mapping studies assign the human nebulette gene to chromosome 10p12, whereas the nebulin gene has been previously assigned to 2q21. Evolutionary constraints appear to have maintained identical modular arrangements in these two independent genes. Comparison of nebulin and nebulette cDNAs demonstrates that a subgroup of repeats within the C terminal regions is regulated tissue-specifically and stage-dependently during development of both molecules. This leads to a substantial diversity of nebulin and nebulette isoforms. Their further study is likely to provide insights into how they contribute to the molecular diversity of Z-discs from different muscle tissues and fiber types. PMID- 9733645 TI - Efficient display of an HCV cDNA expression library as C-terminal fusion to the capsid protein D of bacteriophage lambda. AB - We describe the construction and characterization of a hepatitis C virus (HCV) cDNA expression library displayed as a fusion to the carboxy terminus of the capsid protein D of bacteriophage lambda. cDNA inserts were obtained by tagged random-priming of the HCV genome and cloned into a lambda vector from which chimeric phage bearing both wild-type D protein and D fusion products on the capsid surface were produced. The resulting library was affinity-selected with anti-HCV human monoclonal antibodies recognizing linear or conformational epitopes, and human sera from HCV-infected patients. Selection was monitored by immuno-screening experiments, ELISA, and sequence analysis of positive clones. The performance of this library was compared with two additional HCV cDNA display libraries generated as N-terminal fusions to the III and VIII capsid proteins of filamentous phage M13. The results obtained demonstrate the great potential of the lambda display system for constructing complex cDNA libraries for natural ligand discovery. PMID- 9733646 TI - Structure of a filamentous phosphoglycoprotein polymer: the secreted acid phosphatase of Leishmania mexicana. AB - The insect stage of the protozoan parasite Leishmania mexicana secretes a filamentous acid phosphatase (secreted acid phosphatase, SAP), a polymeric phosphoglycoprotein. The wild-type (wt) SAP filament is a copolymer composed of two related gene products SAP1 and SAP2, which are identical in the enzymatically active NH2-terminal domain and the COOH-terminal domain, but differ in the length of a highly glycosylated Ser/Thr-rich repeat region (32 amino acids and 383 amino acids, respectively) which is located between these domains. When expressed separately, full length SAP1, SAP2, or the NH2-terminal domain alone, are able to assemble into filaments. The Ser/Thr-rich region is the exclusive target for a novel type of O-glycosylation via phosphoserines. By using glycerol spraying/low angle rotary metal shadowing and labelling with monoclonal antibodies it is demonstrated that the repetitive region adopts an extended conformation forming side arms which project radially from the filament core and terminate with the COOH-terminal domain. The length of the side arms of SAP1 and SAP2 (20 nm and 90 nm, respectively) corresponds to the predicted length of the Ser/Thr-rich repeat region of SAP1 and SAP2. Mass determination by scanning electron microscopy (STEM) shows that one morphologically defined globular particle of the filament core is a polypeptide dimer. We propose a model for the filament core, in which the globular NH2-terminal SAP domains form one strand composed of polypeptide dimers or two tightly associated strands of monomers which may twist into a double helix, similar to actin filaments. The highly O-glycosylated side arms project from the filament core conferring an overall bottle-brush-like appearance. The L. mexicana SAP is compared to SAPs secreted by the closely related species L. amazonensis and L. donovani. PMID- 9733647 TI - GlnK, a PII-homologue: structure reveals ATP binding site and indicates how the T loops may be involved in molecular recognition. AB - GlnK is a recently discovered homologue of the PII signal protein, an indicator of the nitrogen status of bacteria. PII occupies a central position in the dual cascade that regulates the activity of glutamine synthetase and the transcription of its gene. The complete role of Escherichia coli GlnK is yet to be determined, but already it is known that GlnK behaves like PII and can substitute for PII under some circumstances thereby adding to the subtleties of nitrogen regulation. There are also indications that the roles of the two proteins differ; the expression of PII is constitutive while that of GlnK is linked to the level of nitrogen in the cell. The discovery of GlnK begs the question of why E. coli has both GlnK and PII. Clearly, the structural similarities and differences of GlnK and PII will lead to a better understanding of how PII-like proteins function in E. coli and other organisms. We have crystallised and solved the X-ray structure of GlnK at 2.0 A resolution. The asymmetric unit has two independent copies of the GlnK subunit and both pack around 3-fold axes to form trimers. The trimers have a barrel-like core with recognition loops (the T-loops) that protrude from the top of the molecule. The two GlnK molecules have similar core structures to PII but differ significantly at the C terminus and the loops. The T-loops of the two GlnK molecules also differ from each other; one is disordered while the conformation of the other is stabilised by lattice contacts. The conformation of the ordered T-loop of GlnK differs from that observed in the PII structure despite the fact that their sequences are very similar. The structures suggest that the T-loops do not have a rigid structure and that they may be flexible in solution. The presence of a turn of 310 helix in the middle of the T-loop suggests that secondary structure could form when it interacts with soluble receptor enzymes.Co-crystals of GlnK and ATP were used to determine the structure of the complex. In these crystals, GlnK occupies a position of 3-fold symmetry. ATP binds in a cleft on the side of the molecule. The cleft is suitably positioned for ATP to influence the flexible T-loops. It is found at the junction of two beta sheets and is formed by two peptides one of which contains a variant of the "Gly-loop" found in other mononucleotide binding proteins. This sequence, Thr-Gly-X-X-Gly-Asp-Gly-Lys-Ile-Phe, forms part of the B-loop and is conserved in a wide variety of organisms that include bacteria, algae and archeabacteria. This sequence is more highly conserved than the functional T-loop, suggesting that ATP has an important role in PII-like proteins. PMID- 9733648 TI - The structure of a trimeric archaeal adenylate kinase. AB - The adenylate kinase from the hyperthermophilic archaean species Sulfolobus acidocaldarius has been cloned, expressed in Escherichia coli, purified and crystallized. The crystal structure was elucidated by multiple isomorphous replacement and non-crystallographic density averaging. The structure was refined at 2.6 A (1 A=0.1 nm) resolution. The enzyme is trimeric, in contrast to previous solution measurements that suggested a dimeric structure, and in contrast to the vast majority of adenylate kinases, which are monomeric. In large parts of each subunit the chain fold resembles the known enzyme structure from eubacteria and eukaryotes although the sequence homology is negligible. Since the asymmetric unit contains two trimers with and without bound AMP at the AMP sites and with an ADP at one of the six ATP sites, the analysis shows the enzyme in several states. The conformational differences between these states resemble those of other adenylate kinases. Because of sequence homology, the structure presented provides a good model for the methanococcal adenylate kinases. PMID- 9733649 TI - The dependence of chemical exchange on boundary selection in a fibronectin type III domain from human tenascin. AB - The third fibronectin type III domain from human tenascin adopts a compact beta sandwich fold. Its boundaries were originally selected to encode a 90-residue domain (TNfn31-90). We conclude that the dynamic properties of TNfn3 are more accurately represented when the C terminus is extended by the two naturally succeeding residues. Longitudinal (R1) and transverse (R2) 15N relaxation rates, and ?1H-15N? NOE enhancements at pH 4.9 and 300 K are presented for TNfn31-90 and TNfn31-92, the extended form, at two field strengths (11.74 and 14.10 T). Nearly identical results confirm their similar motional properties over a broad range of timescales. However, a number of residues near the C terminus in TNfn31-90 exhibit elevated transverse relaxation rates and broadened signals in 1H-15N HSQC spectra. Explicit rates of chemical exchange for five residues in TNfn31-90 were determined by measuring transverse relaxation rates in a series of CPMG experiments with spin-echo refocusing delays increasing from 311 to 1436 micros. Calculated exchange rates average 1000(+/-311) s-1, with individual uncertainties near 20%. Homonuclear TOCSY experiments collected between pH 4 and 7 reveal the coincident titration of two acidic clusters in TNfn31-90 at pH 5. 64(+/-0.47). The repulsive electrostatic interaction of the C-terminal carboxylate with one of these clusters may promote chemical exchange in the shorter domain. Additionally, NOE and chemical shift data suggest hydrogen bond formation between the added residues and adjacent loops. The data affirm the importance of judiciously selecting domain boundaries prior to the characterization of molecular properties. PMID- 9733650 TI - A model of Cdc25 phosphatase catalytic domain and Cdk-interaction surface based on the presence of a rhodanese homology domain. AB - Mammalian Cdc25 phosphatase is responsible for the dephosphorylation of Cdc2 and other cyclin-dependent kinases at Thr14 and Tyr15, thus activating the kinase and allowing cell cycle progression. The catalytic domain of this dual-specificity phosphatase has recently been mapped to the 180 most C-terminal amino acids. Apart from a CX3R motif, which is present at the active site of all known tyrosine phosphatases, Cdc25 does not share any obvious sequence similarity with any of those enzymes. Until very recently, the Cdc25 family was the only subfamily of tyrosine phosphates for which no three-dimensional structural data were available. Using the generalized profile technique, a sensitive method for sequence database searches, we found an extended and highly significant sequence similarity between the Cdc25 catalytic domain and similarly sized regions in other proteins: the non-catalytic domain of two distinct families of MAP-kinase phosphates, the non-catalytic domain of several ubiquitin protein hydrolases, the N and C-terminal domain of rhodanese, and a large and heterogeneous groups of stress-response proteins from all phyla. The relationship of Cdc25 to the structurally well-characterized rhodanese spans the entire catalytic domain and served as template for a structural model for human Cdc25a, which is fundamentally different from previously suggested models for Cdc25 catalytic domain organization. The surface positioning of subfamily-specific conserved residues allows us to predict the sites of interaction with Cdk2, a physiological target of Cdc25a. Based on the results of this analysis, we also predict that the budding yeast arsenate resistance protein Acr2 and the ORF Ygr203w encode protein phosphatases with catalytic properties similar to that of the Cdc25 family. Recent determination of the crystal structure of the Cdc25a catalytic domain supports the validity of the model and demonstrates the power of the generalized sequence profile technique in homology-based modeling of the three-dimensional structure of a protein having a weak but significant sequence similarity with a structurally characterized protein. PMID- 9733652 TI - Life's third domain (Archaea): an established fact or an endangered paradigm? AB - The three-domain proposal of Woese et al. (Proc. Natl. Acad. Sci. USA 87, 4576 (1990)) divides all living organisms into three primary groups or domains named Archaea (or archaebacteria), Bacteria (or eubacteria), and Eucarya (or eukaryotes), with Eucarya being relatives (or descendants) of Archaea. Although this proposal is currently widely accepted, sequence features and phylogenies derived from many highly conserved proteins are inconsistent with it and point to a close and specific relationship between archaebacteria and gram-positive bacteria, whereas gram-negative bacteria are indicated to be phylogenetically distinct. A closer relationship of archaebacteria to gram-positive bacteria in comparison to gram-negative bacteria is generally seen for the majority of the available gene/protein sequences. To account for these results, and the fact that both archaebacteria and gram-positive bacteria are prokaryotes surrounded by a single cell membrane, I propose that the primary division within prokaryotes is between Monoderm prokaryotes (surrounded by a single membrane) and Diderm prokaryotes (i.e., all true gram-negative bacteria containing both an inner cytoplasmic membrane and an outer membrane). This proposal is consistent with both cell morphology and signature sequences in different proteins. Protein phylogenies and signature sequences also show that all eukaryotic cells have received significant gene contributions from both an archaebacterium and a gram- negative eubacterium. Thus, the hypothesis that archaebacteria and eukaryotes shared a common ancestor exclusive of eubacteria, or that the ancestral eukaryotic cell directly descended from an archaea, is erroneous. These results call into question the validity of the currently popular three-domain proposal and the assignment of a domain status to archaebacteria. A new classifica- tion of organisms consistent with phenotype and macromolecular sequence data is proposed. PMID- 9733653 TI - A reconsideration of Galton's problem (using a two-sex population). AB - The main purposes of this paper are to promote and expound the bisexual Galton Watson branching process as a relevant model for the consideration of Francis Galton's problem regarding the extinction of surnames of "men of note." A scheme for adapting the bisexual process to consider Galton's problem is introduced. A necessary and sufficient condition for the certain extinction of a male-induced property in a two-sex species is presented. An approach for calculating the extinction of a male-generated characteristic in the two-sex species is proposed. That approach is then used to find the probability of the extinction of surnames in a bisexual population for Alfred Lotka's data based on an United States Census. Finally, these results are then compared with the classic extinction probabilities (from Lotka) associated with the traditional Galton-Watson branching process using asexual reproduction. PMID- 9733654 TI - Quantifying the intrinsic transmission dynamics of tuberculosis. AB - Previously we have formulated transmission models of untreated tuberculosis epidemics (Blower et al., Nature, Medicine 1 (1995), 815-821); in this paper, we present time-dependent uncertainty and sensitivity analyses in order to quantitatively understand the transmission dynamics of tuberculosis epidemics in the absence of treatment. The time-dependent uncertainty analysis enabled us to evaluate the variability in the epidemiological outcome variables of the model during the progression of a tuberculosis epidemic. Calculated values (from the uncertainty analysis) for the disease incidence, disease prevalence, and mortality rates were approximately consistent with historical data. The time dependent sensitivity analysis revealed that only a few of the model's input parameters significantly affected the severity of a tuberculosis epidemic; these parameters were the disease reactivation rate, the fraction of infected individuals who develop tuberculosis soon after infection, the number of individuals that an infectious individual infects per year, the disease death rate, and the population recruitment rate. Our analysis demonstrates that it is possible to improve our understanding of the behavior of tuberculosis epidemics by applying time-dependent uncertainty and sensitivity analysis to a transmission model. PMID- 9733655 TI - Stable equilibria in multilocus genetic systems: a statistical investigation. AB - A data base of gametic distributions at a stable equilibrium for genetic systems with up to five diallelic loci was created by numerically iterating equations for the dynamics of gametic frequencies in multilocus systems under selection. For a given number of loci, iterations were conducted for 4000 random sets of genotypic fitnesses, 6 values of recombination, and 10 different initial distributions. The data base was used to investigate the following properties of stable equilibria maintaining a polymorphism in a given number of loci that are expected a priori, i.e., without any constraints on fitnesses of genotypes: probability for a fitness set to yield a such equilibrium; probability for a random trajectory to converge to a such equilibrium; genetic load at a such equilibrium. The expected number of simultaneously stable equilibria, and the fraction of genome maintained polymorphic were also investigated as well as some parameters expected at an equilibrium maintaining all loci polymorphic. One of the most important findings is that multilocus genetic systems have a potential for maintaining a polymorphism in a large number of loci under selection without an input of new genetic variation. PMID- 9733656 TI - Signalling among relatives. II. Beyond the tower of Babel. AB - Models of costly signalling are commonly employed in evolutionary biology in order to explain how honest communication between individuals with conflicting interests can be stable. These models have focused primarily on a single type of honest signalling equilibrium, the separating equilibrium in which any two different signallers send distinct signals, thereby providing signal receivers with complete information. In this paper, we demonstrate that in signalling among relatives (modelled using the Sir Philip Sidney game), there is not one but a large number of possible signalling equilibria, most of which are pooling equilibria in which different types of signallers may share a common signal. We prove that in a general Sir Philip Sidney game, any partition of signallers into equi-signalling classes can have a stable signalling equilibrium if and only if it is a contiguous partition, and provide examples of such partitions. A similar (but slightly stricter) condition is shown to hold when signals are transmitted through a medium with signalling error. These results suggest a solution to a problem faced by previous signalling theory models: when we consider the separating equilibrium, signal cost is independent of the frequency of individuals sending that signal and, consequently, even very rare signaller types can drastically affect signal cost. Here, we show that by allowing these rare signallers to pool with more common signallers, signal cost can be greatly reduced. PMID- 9733658 TI - Dynamics of transposable elements in metapopulations: a model of P element invasion in Drosophila. AB - Work on how transposable elements are maintained and spread by virtue of their transposition processes have produced many theoretical studies of their evolutionary dynamics. But recent studies, which have experimentally identified some of these mechanisms, have not been taken into account. We present an integrated model of P transposable element regulation. It includes, at an individual level, the various mechanisms of regulation and the transposition events, that have been experimentally identified, recording specifically the chromosomal localisations of the inserted copies. It attempts to define the minimum conditions for explaining the regulation and spread of the P transposable element in Drosophila melanogaster natural populations. One test of this model is that it must explain the different population states found in the wild. A program that simulates the changes in Drosophila populations during the invasion of P elements was developed; the simulated populations were then compared to natural population data at the molecular and genetic levels. The model was validated by testing the dynamics of P element invasion in populations. It could explain the different natural population states with a recurrent invasion process. The simulations show that migration reduces the total number of copies, increases the number of defective copies, decreases P-activity and increases P-susceptibility, shifting equilibrium states from P to M'. They also show that the copies determining P-cytotype regulation spread faster by selection when located on the X chromosome. This result could account for the unexplained accumulation of P copies on the X chromosomes of some natural populations. Moreover the simulations predict a novel equilibrium state, called P', not yet characterized in natural populations but that can be found in natural population data. PMID- 9733657 TI - Restrictions on components of variance for epistatic models. AB - If a disease is caused by several loci, then the additive variance at each locus may represent only small portions of the total genetic variance, while epistatic variance components may explain a significant amount of the total genetic variance. In this paper we first give simple general formulations to derive all the components of total genetic variance in a random sample for any multilocus model. We then derive these components for a series of fifteen models that have been proposed as being the two-allele two-locus models most likely for disease. We discuss the restrictions and limitations on the penetrance and the gene frequencies, implied by the disease prevalence, for each model. We investigate the relative magnitudes of the components of variance for the various models and show that in six of the models one or other of the epistatic variance components can be larger than each of the other components. It is suggested that investigations be undertaken to develop appropriate sampling and analytical techniques to detect these variance components by linkage analysis. PMID- 9733659 TI - The N-glucuronidation of xenobiotics. An aspet-supported symposium held at the 1996 faseb meeting in washington, dc PMID- 9733660 TI - N+-glucuronidation, a common pathway in human metabolism of drugs with a tertiary amine group. AB - Glucuronidation of either an aliphatic or aromatic tertiary amine group in a molecule results in a quaternary ammonium-linked glucuronide metabolite (i.e. N+ glucuronide). The development of sound information on N+-glucuronide metabolites, including their characterization, has been slow. In part, this is because the presence of both the carboxylic acid group and cationic center in their structure imparts physiochemical properties such that procedures used in their analysis, including extraction, require judicious selection. The techniques used in the identification of N+-glucuronide metabolites and those metabolites identified in human urine are the focus of this review. Especially useful in their identification are the availability of an authentic synthetic sample and the use of mass spectrometry and nuclear magnetic resonance (NMR) techniques that, in the first instance, involve atmospheric pressure ionization or fast atom bombardment modes of ionization and high-resolution 1H NMR. More than 30 N+-glucuronide metabolites of xenobiotics have been identified in human urine. In particular, N+ glucuronidation is a common phenomenon in the metabolism of H1 antihistamine and antidepressant drugs with an aliphatic tertiary amine group. Those marketed drugs in which the reported N+-glucuronide mean urinary excretion of the orally administered dose exceeds 10% include cyclizine, cyclobenzaprine, cyproheptadine, dothiepin, doxepin, ketotifen, lamotrigine, mianserin, and tioconazole. The pharmacological importance of N+-glucuronidation has not been clarified. PMID- 9733661 TI - Species differences in N-glucuronidation. AB - Glucuronidation of amines has been shown to exhibit species differences in vitro and in vivo. Substrates for N-glucuronidation can be classified according to the chemical structures of the resulting glucuronides into two groups: compounds that form non-quaternary N-conjugates, and those that form the quaternary counterparts. For compounds of the former class-such as sulfonamides, arylamines, and alicyclic, cyclic, and heterocyclic amines-species differences appear to be less striking and are of a quantitative nature. No one common laboratory animal species used routinely in metabolism research (e.g. rat, mouse, dog, non-human primate, rabbit, and guinea pig) has been shown to be deficient in N glucuronidation when all of the substrates studied and reported are taken into consideration. The ability of a species to form N-glucuronides is compound dependent, although rabbit and guinea pig appear to exhibit the highest capacity for this bioconjugation among preclinical species. For tertiary amines, most notably the tricyclic antidepressant and antihistamine drugs, N-glucuronidation is commonly observed in non-human primates and man. There are examples, however, of quaternary glucuronidation occurring in lower animal species. In exploring species differences in amine conjugation in vivo, it is noted that the apparent absence of N-glucuronides in animal urine may not reflect the inability of that species to form such conjugates, since the N-glucuronides may be excreted in bile. Problems such as degradation or low recoveries commonly encountered in isolation and identification of in vivo metabolites further complicate the interpretation of data. Because of the wide range of pKa values exhibited by various classes of amines, caution also should be exercised for in vitro studies since incubation conditions for N-glucuronidation often are substrate- and species-dependent. Explanations for the species differences observed in N glucuronidation appear to be emerging as rapid advances are made in the understanding of the glucuronosyltransferases at the molecular level. More information, however, remains to be gathered from the glucuronosyltransferase genes of animal species other than humans before a better understanding of species differences in N-glucuronidation can be achieved. PMID- 9733662 TI - Olanzapine 10-N-glucuronide. A tertiary N-glucuronide unique to humans. AB - In humans, a major metabolite of the atypical antipsychotic olanzapine in the plasma and in the urine was found to be an N-glucuronide. Unexpectedly, the glucuronic acid moiety was linked through a nitrogen of the benzodiazepine nucleus of olanzapine by way of a secondary amine linkage, rather than through a nitrogen on the piperazine substituent of the nucleus, to give a quaternary ammonium glucuronide. Derivatization with phenylisothiocyanate to yield a thiourea adduct indicated that conjugation occurred via a secondary amine. Subsequently, mass spectrometry and nuclear magnetic resonance studies with the isolated metabolite and later with the synthesized metabolite indicated that the glucuronide was linked at the 10- position of olanzapine. This phase 2 metabolite was only detected in the plasma and urine of human subjects and not in mice, rats, or monkeys; a trace of this metabolite was detected in dog urine. The N-10 glucuronide was resistant to enzymatic and base hydrolysis but was cleaved under acidic conditions. Formation of an N-glucuronide metabolite directly with the benzodiazepine nucleus has not previously been reported. PMID- 9733663 TI - N-glucuronidation of benzidine and its metabolites. Role in bladder cancer. AB - Workers exposed to high levels of benzidine have a 100-fold increased incidence of bladder cancer. This review evaluates the overall metabolism of benzidine to determine pathways important to initiation of bladder cancer. Upon incubation of benzidine with liver slices from rats, dogs, and humans, different proportions of this diamine were N-acetylated and N-glucuronidated. With dogs, a non-acetylator species, N-glucuronidation was the major pathway. In contrast, little glucuronidation was observed in rats with N, N'-diacetylbenzidine, the major metabolite of benzidine. Human liver slices demonstrated both extensive N acetylation and N-glucuronidation. Differences between rats and humans were attributed to rapid deacetylation by human liver with N-acetylbenzidine rather than an accumulation of N, N'-diacetylbenzidine. N-Acetylbenzidine oxidative metabolism was also observed. The acid lability of glucuronide products of benzidine, N-acetylbenzidine, and oxidation products of N-acetylbenzidine metabolism was assessed. N-Glucuronides of benzidine, N-acetylbenzidine, and N' hydroxy-N-acetylbenzidine were acid-labile, with the latter having a much longer half-time than the former two glucuronides. Because bladder epithelium contains relatively high levels of prostaglandin H synthase and not cytochrome P450, the peroxidative metabolism of N-acetylbenzidine was assessed. N'-(3'-Monophospho deoxyguanosin-8-yl)-N-acetylbenzidine was the only DNA adduct detected. This adduct is also the major adduct detected in bladder cells from workers exposed to benzidine. In urine from these workers, an inverse relationship between urine pH and levels of free (unconjugated) benzidine and N-acetylbenzidine was observed. A similar inverse relationship was observed for urine pH and levels of bladder cell N'-(3'-monophospho-deoxyguanosin-8-yl)-N-acetylbenzidine. These results suggest multiple pathways (acetylation, glucuronidation, peroxidation) in multiple organs (liver, blood, kidney, bladder) are important in benzidine-induced bladder cancer. PMID- 9733664 TI - Glucuronidation of amine substrates by purified and expressed UDP glucuronosyltransferase proteins. AB - Conjugation of many primary, secondary, and tertiary amine-containing xenobiotics with glucuronic acid can result in the formation of N-glucuronide metabolites. For carcinogenic arylamines and their N-hydroxylated metabolites, N glucuronidation can result in the formation of either inactive metabolites or labile conjugates, which can be transported to their target tissue (urinary bladder) where they may be converted to reactive metabolites. Drugs with primary amine (e.g. dapsone) or secondary amine moieties (e.g. sulfadimethoxine and clozapine) can also be metabolized to N-glucuronides. The metabolism of a number of tertiary amine-containing pharmacological agents to quaternary ammonium-linked glucuronides represents a unique and important metabolic pathway for these compounds that is highly species-dependent. This review summarizes our present knowledge of the uridine diphosphate (UDP)-glucuronosyltransferase enzymes involved in catalyzing N-glucuronide formation. Of the more than 30 UDP glucuronosyltransferases that have been purified or cloned and expressed, many catalyze N-glucuronide formation for primary and secondary amine substrates. In contrast, only human UDP-glucuronosyltransferases 1A3 and 1A4 have been shown to catalyze quaternary ammonium-linked glucuronide formation for aliphatic tertiary amines. The structure of the UGT1 gene complex is highly conserved across species, and it appears that a mutation in the first exon encoding UDP glucuronosyltransferase 1A4, resulting in a pseudo-gene, may explain the inability of some species to form quaternary ammonium-linked glucuronides. PMID- 9733665 TI - Human cytochrome P450-catalyzed conversion of the proestrogenic pesticide methoxychlor into an estrogen. Role of CYP2C19 and CYP1A2 in O-demethylation. AB - 1,1,1-Trichloro-2,2-bis(4-methoxyphenyl)ethane (methoxychlor) is a widely used pesticide that is pro-estrogenic. We have elucidated the human cytochrome P450 enzymes responsible for conversion of methoxychlor into its major metabolite, the mono-O-demethylated derivative (mono-OH-M) that is estrogenic. Incubation of methoxychlor with microsomes from insect cells overexpressing either CYP1A2, CYP2C18, or CYP2C19 yielded mono-OH-M with turnover numbers of 14.9, 15.5, and 39.1 nmol/min/nmol of P450, respectively. CYP2B6 and CYP2C9 were much less active. Incubations with purified CYP2C19 and CYP2C18 resulted in formation of mono-OH-M, and also the bis-demethylated metabolite. Co-incubation of liver microsomes with methoxychlor and various P450 isoform-selective inhibitors suggested involvement of several P450s in mono-O-demethylation, including CYP1A2, CYP2A6, CYP2C9, and CYP2C19. A role for CYP2C19, CYP1A2, and CYP2A6 was also indicated by multivariate regression analysis of the mono-O-demethylase activity in a panel of human liver microsomes characterized for isoform-specific catalytic activities (R2 = 0.96). Based on the totality of the evidence, CYP2C19 appears to be the major catalyst of methoxychlor mono-O-demethylation. However, in individuals lacking functional CYP2C19 (e.g. the "poor metabolizer" phenotype), CYP1A2 may play the predominant role. CYP2A6, CYP2C9, and CYP2B6 probably contribute to a lesser extent. Although CYP2C18 is an efficient methoxychlor demethylase, its expression in liver is reportedly low or absent, suggesting a negligible role for this enzyme in methoxychlor metabolism. Lengthy incubations of liver microsomes with methoxychlor produced other secondary and tertiary metabolites. Efficient conversion of methoxychlor to estrogenic mono-OH-M by liver microsomes suggests that methoxychlor has the potential to be estrogenic in humans, as observed in several animal species. PMID- 9733666 TI - Interaction of terfenadine and its primary metabolites with cytochrome P450 2D6. AB - The substrate structure-activity relationships described for the major human drug metabolizing cytochrome P450 (P450 or CYP) enzymes suggest that the H1 receptor antagonist terfenadine could interact with CYP2D6 either as a substrate or as an inhibitor, in addition to its known ability to act as a substrate for CYP3A4. Based on this substrate structure-activity relationship, computer modeling studies were undertaken to explore the likely interactions of terfenadine with CYP2D6. An overlay of terfenadine and dextromethorphan, a known substrate of CYP2D6, showed that it was possible to superimpose the site of hydroxylation (t butyl group) and the nitrogen atom of terfenadine with similar regions in dextromethorphan. These observations were substantiated by the ease of docking of terfenadine into a protein model of CYP2D6. Experimentally, terfenadine inhibited CYP2D6 activity in human liver microsomes with an IC50 of 14-27 microM, depending on the CYP2D6 substrate used. The inhibition of CYP2D6 was further defined by determining the Ki for terfenadine against bufuralol 1'-hydroxylase activity in four human livers. Terfenadine inhibited bufuralol 1'-hydroxylase activity with a Ki of approximately 3.6 microM. The formation of the hydroxylated metabolite (hydroxyterfenadine) in microsomes prepared from human liver and specific P450 cDNA-transfected B lymphoblastoid cells indicated that only CYP2D6 and CYP3A4 were involved in this transformation. As expected, the rate of formation was greatest with CYP3A4 (Vmax = 1257 pmol/min/nmol of P450), with CYP2D6 forming the metabolite at a 6-fold lower rate (Vmax = 206 pmol/min/nmol of P450). The two enzymes had similar KM values (9 and 13 microM, respectively). These data indicate that, as predicted from modeling studies, terfenadine has the structural features necessary for interaction with CYP2D6. PMID- 9733667 TI - Comparative pharmacokinetics of oral and intravenous ifosfamide/mesna/methylene blue therapy. AB - Oral treatment with ifosfamide results in dose-limiting encephalopathy. Methylene blue is effective in reversal and prophylaxis of this side effect. In the present study, the pharmacokinetics of ifosfamide after iv and po therapy in combination with prophylactic administration of methylene blue were investigated. Nine patients with metastatic non-small cell lung cancer were treated by a combination of ifosfamide (3 days), sodium 2-mercaptoethane sulfonate (4 days), and etoposide (8 days). Cycles were repeated every 28 days. Ifosfamide was administered orally, with the exception of one of the first two cycles, when it was administered as a short infusion (randomly assigned). The patients received methylene blue in doses of 50 mg po 3 times daily; an initial dose of 50 mg was given the evening before chemotherapy. Urine samples were collected over the entire treatment period, and concentrations of ifosfamide and its major metabolite, 2-chloroethylamine, were measured by gas liquid chromatography. By the same technique, 2- and 3 dechloroethylifosfamide were determined in plasma and urine. Overall alkylating activity in urine was assayed by reaction of the alkylating metabolites with 4 (4'-nitrobenzyl)-pyridine. The chemotherapeutic regimen was well-tolerated by all of the patients studied. There was no evidence of a shift in the metabolic pattern dependent on the route of administration. From the data, we conclude that methylene blue has a neuroprotective effect and that the pharmacokinetics of ifosfamide are not influenced by its comedication. PMID- 9733668 TI - Metabolism of clenbuterol in rats. AB - The metabolic fate of [14C]clenbuterol was studied in male and female Wistar rats. After a single oral dose of 200 microgram/kg [14C]clenbuterol, in an 8-day study period, approximately 60% of the radioactivity was eliminated in urine; 20 and 30% of the radioactivity was excreted in feces by male and female rats, respectively. HPLC coupled to on-line radioactivity detection allowed the separation and quantitation of clenbuterol metabolites, some of which were found to be poorly stable in urine. Most of the urinary and fecal metabolites of clenbuterol were isolated and identified using various MS techniques. Analytical methods were also developed to establish the metabolic profiles in feces and tissues, up to 72 hr after clenbuterol administration. Clenbuterol was mainly metabolized by N-dealkylation (secondary amine), as well as N-oxidation and sulfate conjugation (primary amine). Gender-related differences in the rates of clenbuterol N-dealkylation were observed. 4-N-Hydroxylamine was the major metabolite detected in urine, whereas more than one half of the radioactivity in feces was associated with clenbuterol sulfamate. PMID- 9733669 TI - Deposition and retention of radiolabeled serum constituents in hair after systemic administration. AB - To investigate the chemical mechanisms involved in the accumulation of drugs or other compounds in hair, we examined the deposition of radiolabeled serum constituents in the hair of BALB/c (albino) and C57 (pigmented) mice. The extents of in vivo incorporation of a normal serum cation (45Ca2+), a serum anion (36C1 ), a neutral constituent ([14C]urea), and a structural component of hair ([35S]cysteine) were studied to provide a reference framework for the examination of foreign substances deposited in hair from serum. The use of two mouse strains allowed evaluation of the effect of hair pigmentation on levels of accumulation. Additionally, the endogenous contents of Mg2+, Na+, and K+ (measured by inductively coupled plasma-atomic emission spectroscopy) were determined, as was their stability to removal. Hair concentrations of isotopes were calculated from mean specific activities determined over the treatment period and corrected for quenching and decay. 45Ca2+ accumulation (500 ng/mg of hair in C57 mice and 25 ng/mg of hair in BALB/c mice) was unaffected by 24-hr phosphate buffer extraction. Of the [14C]urea accumulated (3500 ng/mg in C57 and BALB/c mice), 50% was removed by 24-hr extraction in phosphate buffer. Of the 36C1- accumulated (65 ng/mg in C57 mice and 30 ng/mg in BALB/c mice), one half was removed by 24-hr extraction in phosphate buffer. The accumulated [35S]cysteine (210 ng/mg in C57 mice and 110 ng/mg in BALB/c mice) could not be removed. Endogenous Mg2+ (350 ng/mg in C57 mice and 75 ng/mg in BALB/c mice) was stable to 24-hr extraction with phosphate buffer. K+ (2500 ng/mg) and Na+ (400 ng/mg) concentrations were approximately equal in the two strains and were largely extractable. Based on the accumulation of a neutral serum constituent (urea), the data suggest that factors other than ionic binding are important in the deposition of circulating molecules into hair. The extent and reversibility of ionic binding are dependent on the chemical nature of the binding substance. The presence of hair pigmentation greatly increased the accumulation of 45Ca2+, 36C1-, and [35S]cysteine. These data suggest a multicompartmental nature of drug storage in hair. PMID- 9733670 TI - In vivo metabolism and disposition of the nephrotoxicant N-(3, 5 dichlorophenyl)succinimide in Fischer 344 rats. AB - N-(3,5-Dichlorophenyl)succinimide (NDPS) was originally developed as an agricultural fungicide. Previous work indicated that NDPS-induced renal damage in rats is metabolism-dependent and that hydroxylated metabolites might be involved in the nephrotoxic response. In this study, the disposition and nephrotoxicity of [14C]NDPS at two time points (3 and 24 hr) and three doses (0.2, 0.4, and 0.6 mmol/kg) were examined in male Fischer 344 rats. At 3 hr, only approximately 6.0% of the administered dose (0.6 mmol/kg) had been excreted. Elimination was nearly complete by 24 hr, except at the highest dose. Urinary elimination far exceeded fecal elimination at all doses. The urinary metabolites were identified as N-(3, 5-dichlorophenyl)succinamic acid, N-(3, 5-dichlorophenyl)-2-hydroxysuccinamic acid, N-(3, 5-dichlorophenyl)-3-hydroxysuccinamic acid, and N-(3, 5 dichlorophenyl)malonamic acid. N-(3, 5Dichlorophenyl)-3-hydroxysuccinamic acid had not been previously detected in vivo. The same metabolites were also detected in the feces, blood, liver, and kidneys of rats. In addition, two novel in vivo NDPS metabolites were detected in liver and kidney homogenates. These metabolites were tentatively identified as N-(3, 5-dichlorophenyl)-2-hydroxysuccinimide and N (3, 5-dichloro-4-hydroxyphenyl)succinamic acid. Dose-dependent increases in blood urea nitrogen levels, diuresis, proteinuria, glucosuria, and covalent protein adducts correlated with increases in oxidative metabolism. Rapid NDPS metabolism could help explain the early onset of nephrotoxicity. These studies provide additional evidence for the importance of oxidative metabolism in NDPS-induced kidney damage. PMID- 9733671 TI - Metabolism of 3-butene-1,2-diol in B6C3F1 mice. Evidence for involvement of alcohol dehydrogenase and cytochrome p450. AB - 3-Butene-1,2-diol (BDD), a metabolite of 1,3-butadiene, is rapidly metabolized by B6C3F1 mice at doses ranging from 10 to 250 mg/kg. Calculation of plasma clearance suggested that the kinetics of BDD metabolism were dose-dependent. Clearance varied 5-fold in this dose range. Urinary excretion of BDD was also dose-dependent but did not exceed 5% of the administered dose. A small fraction of the dose (<1%) was excreted as glucuronide or sulfate conjugates. Benzylimidazole, a cytochrome P450 inhibitor, decreased the clearance of BDD (25 mg/kg) by 44%, whereas 4-methylpyrazole, an alcohol dehydrogenase and cytochrome P450 inhibitor, decreased BDD clearance by 82%. BDD administration (250 mg/kg) resulted in depletion of hepatic and renal nonprotein thiols, by 48 and 22%, respectively. Pretreatment of mice with 4-methylpyrazole provided partial protection against depletion of nonprotein thiols, whereas pretreatment with benzylimidazole was ineffective. Incubation of BDD with NADPH and mouse liver microsomes resulted in time-dependent inactivation of p-nitrophenol hydroxylase (PNPH) activity, a marker for cytochrome P450. Inclusion of glutathione, with or without glutathione peroxidase, did not attenuate the inactivation of PNPH, whereas deferoxamine, superoxide dismutase, catalase, and mannitol provided modest protection. These results are consistent with suicide inhibition of PNPH by BDD, with a minor role for reactive oxygen species in the loss of PNPH. Treatment of mice with BDD (250 mg/kg) inactivated hepatic microsomal PNPH activity by 50% after 60 min. These results suggest that BDD is extensively and rapidly metabolized in mice, and they provide evidence for the formation of reactive intermediates that could play a role in the toxicity of 1, 3-butadiene. PMID- 9733672 TI - Covalent sequestration of the nitrogen mustard mechlorethamine by metallothionein. AB - The research reported here demonstrates covalent binding to the metal-binding protein metallothionein (MT) by the therapeutic nitrogen mustard mechlorethamine. The most surprising aspect of this interaction is the selectivity of the alkylating agent for specific residues of MT. A combination of MS and proteolytic and enzymatic methods was used to deduce specific locations of mechlorethamine alkylation. These experiments indicated that alkylation occurs predominantly in the carboxyl domain of MT, with one molecule of mechlorethamine covalently cross linking two cysteine residues. Electrospray MS revealed the retention of all seven metal ions in the cross-linked MT/mechlorethamine adducts, highlighting the uniqueness of this protein. Computerized docking experiments supported the hypothesis that selective binding precedes selective alkylation, and the structure of the drug indicates the minimal structural requirements for this binding. These results support the idea that MT overexpressed in tumor cells contributes to the inactivation of anticancer drugs. PMID- 9733673 TI - In vitro and in vivo metabolism of desogestrel in several species. AB - The metabolism of desogestrel (13-ethyl-11-methylene-18, 19-dinor-17alpha-pregn-4 en-20-yn-17-ol), an orally active progestogen, was studied in vivo after administration of single oral doses to rats and dogs and in vitro using rat, rabbit, dog, and human liver microsomes. Metabolites were isolated and identified by NMR and MS analysis. After oral administration of [3H]desogestrel to rats and dogs, desogestrel was extensively metabolized in both species. Radioactivity was predominantly eliminated in the feces. In rats, desogestrel was metabolized mainly at the C3-, C5-, C11-, and C15-positions. Both in vivo and in vitro, the majority of metabolites were 3alpha-hydroxy,4,5alpha-dihydro derivatives. Other main metabolic routes for desogestrel in rats were 15alpha-hydroxylation and epoxidation of the C11-methylene moiety. In addition to phase I metabolites, glucuronic acid and sulfate conjugates of desogestrel were observed in vivo. In dogs, desogestrel was mainly metabolized at the C3- and C17-positions. In contrast to the rat metabolites, metabolites isolated from dog urine or feces were mainly 3beta-hydroxy,4,5alpha-dihydro derivatives. In most of the metabolites present in dog urine and feces, the five-membered D-ring was expanded to a six-membered D-ring, i.e. D-homoannulation to a 17A-keto-D-homo ring. D-Homo metabolites, which were major metabolites in plasma, urine, and feces of dogs, were not observed in vitro. In dog liver microsomes, the 3-keto metabolite of desogestrel was the major metabolite. Similarly to dog liver microsomes, rabbit and human liver microsomes mainly converted desogestrel to its 3-keto metabolite. Predominant positions for further hydroxylation of the 3-keto metabolite of desogestrel were the C6-position (6beta-hydroxy) and the ethyl substituent at the C13-position, for both species. PMID- 9733674 TI - Pharmacokinetics and blood-brain barrier transport of an anti-transferrin receptor monoclonal antibody (OX26) in rats after chronic treatment with the antibody. AB - Monoclonal antibodies (MAbs) directed against cell surface receptors (e.g. the transferrin receptor or the insulin receptor) on the brain capillary endothelium, which makes up the blood-brain barrier (BBB) in vivo, are brain drug-delivery vectors. When cells are chronically exposed to MAbs in tissue culture, there is often down-regulation of the cell surface receptors. To examine whether similar down-regulation occurs in vivo, rats were chronically treated either with the OX26 murine MAb to the rat transferrin receptor or with a mouse IgG2a isotype control (0.25 mg/kg sc daily for 1 week), and the BBB transport of the OX26 MAb was then measured for both rat brain and liver in vivo. Although this treatment regimen resulted in a 41% increase in the permeability-surface area product for 125I-OX26 MAb transport into rat liver in vivo, there was no significant change in the BBB permeability-surface area product for the OX26 MAb. These studies indicate that repetitive administration of cell surface-specific MAbs does not necessarily result in down-regulation of BBB receptors. PMID- 9733675 TI - Cimetidine sulfoxidation in small intestinal microsomes. AB - In previous studies, sulfoxide metabolite was observed in animal and human intestinal perfusions of cimetidine and other H2-antagonists in vivo. L Methionine, imipramine, and the anionic exchange inhibitor diisothiocyanostilbene 2,2'-disulfonic acid reduced metabolite appearance. A sequence of follow-up studies is underway, for the purpose of assessing the contributions of drug metabolism and drug and metabolite transport to variable drug absorption. In this regard, drug-drug and drug-nutrient interactions represent a primary focus of this research. The S-oxidation of cimetidine in mammalian small intestinal microsomes was studied from three different species and two intestinal regions. Based on preparation activity and tissue availability, the relative contributions of flavin-containing monooxygenases and cytochrome P450 enzymes to cimetidine sulfoxidation were evaluated in rabbit jejunal microsomes. Additional inhibitor studies were carried out to evaluate the role of microsomal cimetidine sulfoxidation in the previous in vivo observations. PMID- 9733676 TI - Impact of culture-independent studies on the emerging phylogenetic view of bacterial diversity. PMID- 9733677 TI - Mutations in Salmonella pathogenicity island 2 (SPI2) genes affecting transcription of SPI1 genes and resistance to antimicrobial agents. AB - The Salmonella typhimurium genome contains two pathogenicity islands (SPI) with genes encoding type III secretion systems for virulence proteins. SPI1 is required for the penetration of the epithelial layer of the intestine. SPI2 is important for the subsequent proliferation of bacteria in the spleens of infected hosts. Although most mutations in SPI2 lead to a strong reduction of virulence, they have different effects in vitro, with some mutants having significantly increased sensitivity to gentamicin and the antibacterial peptide polymyxin B. Previously we showed that certain mutations in SPI2 affect the ability of S. typhimurium to secrete SPI1 effector proteins and to invade cultured eukaryotic cells. In this study, we show that these SPI2 mutations affect the expression of the SPI1 invasion genes. Analysis of reporter fusions to various SPI1 genes reveals highly reduced expression of sipC, prgK, and hilA, the transcriptional activator of SPI1 genes. These observations indicate that the expression of one type III secretion system can be influenced dramatically by mutations in genes encoding a second type III secretion system in the same cell. PMID- 9733678 TI - Characterization of native and recombinant forms of an unusual cobalt-dependent proline dipeptidase (prolidase) from the hyperthermophilic archaeon Pyrococcus furiosus. AB - Proline dipeptidase (prolidase) was purified from cell extracts of the proteolytic, hyperthermophilic archaeon Pyrococcus furiosus by multistep chromatography. The enzyme is a homodimer (39.4 kDa per subunit) and as purified contains one cobalt atom per subunit. Its catalytic activity also required the addition of Co2+ ions (Kd, 0.24 mM), indicating that the enzyme has a second metal ion binding site. Co2+ could be replaced by Mn2+ (resulting in a 25% decrease in activity) but not by Mg2+, Ca2+, Fe2+, Zn2+, Cu2+, or Ni2+. The prolidase exhibited a narrow substrate specificity and hydrolyzed only dipeptides with proline at the C terminus and a nonpolar amino acid (Met, Leu, Val, Phe, or Ala) at the N terminus. Optimal prolidase activity with Met-Pro as the substrate occurred at a pH of 7.0 and a temperature of 100 degrees C. The N-terminal amino acid sequence of the purified prolidase was used to identify in the P. furiosus genome database a putative prolidase-encoding gene with a product corresponding to 349 amino acids. This gene was expressed in Escherichia coli and the recombinant protein was purified. Its properties, including molecular mass, metal ion dependence, pH and temperature optima, substrate specificity, and thermostability, were indistinguishable from those of the native prolidase from P. furiosus. Furthermore, the Km values for the substrate Met-Pro were comparable for the native and recombinant forms, although the recombinant enzyme exhibited a twofold greater Vmax value than the native protein. The amino acid sequence of P. furiosus prolidase has significant similarity with those of prolidases from mesophilic organisms, but the enzyme differs from them in its substrate specificity, thermostability, metal dependency, and response to inhibitors. The P. furiosus enzyme appears to be the second Co-containing member (after methionine aminopeptidase) of the binuclear N-terminal exopeptidase family. PMID- 9733679 TI - Mutational inactivation of a gene homologous to Escherichia coli ptsP affects poly-beta-hydroxybutyrate accumulation and nitrogen fixation in Azotobacter vinelandii. AB - Strain DS988, an Azotobacter vinelandii mutant with a reduced capacity to accumulate poly-beta-hydroxybutyrate, was isolated after mini-Tn5 mutagenesis of the UW136 strain. Cloning and nucleotide sequencing of the affected locus revealed a gene homologous to Escherichia coli ptsP which encodes enzyme INtr, a homologue of enzyme I of the phosphoenol pyruvate-sugar phosphotransferase system with an N-terminal domain similar to the N-terminal domain of some NifA proteins. Strain DS988 was unable to grow diazotrophically with 10 mM glucose as a carbon source. Diazotrophic growth on alternative carbon sources such as gluconate was only slightly affected. Glucose uptake, as well as glucose kinase and glucose-6 phosphate-dehydrogenase activities that lead to the synthesis of gluconate-6 phosphate, were not affected by the ptsP mutation. The inability of DS988 to grow diazotrophically in 10 mM glucose was overcome by supplying ammonium or other sources of fixed nitrogen. Acetylene reduction activity but not transcription of the nitrogenase structural gene nifH was shown to be impaired in strain DS988 when it was incubated in 10 mM glucose. The diazotrophic growth defect of DS988 was restored either by increasing the glucose concentration to above 20 mM or by lowering the oxygen concentration. These data suggest that a mutation in ptsP leads to a failure in poly-beta-hydroxybutyrate metabolism and in the respiratory protection of nitrogenase under carbon-limiting conditions. PMID- 9733680 TI - Probing the role of cysteine residues in glucosamine-1-phosphate acetyltransferase activity of the bifunctional GlmU protein from Escherichia coli: site-directed mutagenesis and characterization of the mutant enzymes. AB - The glucosamine-1-phosphate acetyltransferase activity but not the uridyltransferase activity of the bifunctional GlmU enzyme from Escherichia coli was lost when GlmU was stored in the absence of beta-mercaptoethanol or incubated with thiol-specific reagents. The enzyme was protected from inactivation in the presence of its substrate acetyl coenzyme A (acetyl-CoA), suggesting the presence of an essential cysteine residue in or near the active site of the acetyltransferase domain. To ascertain the role of cysteines in the structure and function of the enzyme, site-directed mutagenesis was performed to change each of the four cysteines to alanine, and plasmids were constructed for high-level overproduction and one-step purification of histidine-tagged proteins. Whereas the kinetic parameters of the bifunctional enzyme appeared unaffected by the C296A and C385A mutations, 1,350- and 8-fold decreases of acetyltransferase activity resulted from the C307A and C324A mutations, respectively. The Km values for acetyl-CoA and GlcN-1-P of mutant proteins were not modified, suggesting that none of the cysteines was involved in substrate binding. The uridyltransferase activities of wild-type and mutant GlmU proteins were similar. From these studies, the two cysteines Cys307 and Cys324 appeared important for acetyltransferase activity and seemed to be located in or near the active site. PMID- 9733681 TI - Transcription analysis of two disparate rRNA operons in the halophilic archaeon Haloarcula marismortui. AB - The genome of the halophilic archaeon Haloarcula marismortui contains two rRNA operons designated rrnA and rrnB. Genomic clones of the two operons and their flanking regions have been sequenced, and primary transcripts and processing intermediates derived from each operon have been characterized. The 16S, 23S, and 5S genes from the two operons were found to differ at 74 of 1,472 positions, 39 of 2,922 positions, and 2 of 122 positions, respectively. This degree of sequence divergence for multicopy (paralogous) rRNA genes was 10- to 50-fold or more higher than anticipated. The two operons exhibit other profound differences that include (i) the presence in rrnA and the absence in rrnB of tRNAAla and tRNACys genes in the intergenic and distal regions, respectively, (ii) divergent 5' flanking sequences, and (iii) distinct pathways for processing and maturation of 16S rRNA. Processing and maturation of 16S and 23S rRNA from rrnA operon transcripts and of 23S rRNA from rrnB operon transcripts follow the canonical halophilic pathway, whereas maturation of 16S rRNA from rrnB operon transcripts follows an unusual and different pathway that is apparently devoid of any 5' processing intermediate. PMID- 9733682 TI - Deletion of the alternative sigma factor sigmaB in Staphylococcus aureus reveals its function as a global regulator of virulence genes. AB - A deletion of the sigB operon was constructed in three genetically distinct Staphylococcus aureus strains, and the phenotypes of the resulting mutants were analyzed. Compared to the corresponding wild-type strains, the DeltasigB mutants showed reduced pigmentation, accelerated sedimentation, and increased sensitivity to hydrogen peroxide during the stationary growth phase. A cytoplasmic protein missing in the DeltasigB mutants was identified as alkaline shock protein 23, and an extracellular protein excreted at higher levels in one of the DeltasigB mutants was identified as staphylococcal thermonuclease. Interestingly, most sigB deletion phenotypes were only seen in S. aureus COL and Newman and not in 8325, which was found to contain an 11-bp deletion in the regulator gene rsbU. Taken together, our results show that sigmaB is a global regulator which modulates the expression of several virulence factors in S. aureus and that laboratory strain 8325 is a sigmaB-defective mutant. PMID- 9733684 TI - Phosphorylation of the periplasmic binding protein in two transport systems for arginine incorporation in Escherichia coli K-12 is unrelated to the function of the transport system. AB - In Escherichia coli K-12, the accumulation of arginine is mediated by two distinct periplasmic binding protein-dependent transport systems, one common to arginine and ornithine (AO system) and one for lysine, arginine, and ornithine (LAO system). Each of these systems includes a specific periplasmic binding protein, the AO-binding protein for the AO system and the LAO-binding protein for the LAO system. The two systems include a common inner membrane transport protein which is able to hydrolyze ATP and also phosphorylate the two periplasmic binding proteins. Previously, a mutant resistant to the toxic effects of canavanine, with low levels of transport activities and reduced levels of phosphorylation of the two periplasmic binding proteins, was isolated and characterized (R. T. F. Celis, J. Biol. Chem. 265:1787-1793, 1990). The gene encoding the transport ATPase enzyme (argK) has been cloned and sequenced. The gene possesses an open reading frame with the capacity to encode 268 amino acids (mass of 29.370 Da). The amino acid sequence of the protein includes two short sequence motifs which constitute a well-defined nucleotide-binding fold (Walker sequences A and B) present in the ATP-binding subunits of many transporters. We report here the isolation of canavanine-sensitive derivatives of the previously characterized mutant. We describe the properties of these suppressor mutations in which the transport of arginine, ornithine, and lysine has been restored. In these mutants, the phosphorylation of the AO- and LAO-binding proteins remains at a low level. This information indicates that whereas hydrolysis of ATP by the transport ATPase is an obligatory requirement for the accumulation of these amino acids in E. coli K 12, the phosphorylation of the periplasmic binding protein is not related to the function of the transport system. PMID- 9733683 TI - Topological analysis of DcuA, an anaerobic C4-dicarboxylate transporter of Escherichia coli. AB - Escherichia coli possesses three independent anaerobic C4-dicarboxylate transport systems encoded by the dcuA, dcuB, and dcuC genes. The dcuA and dcuB genes encode related integral inner-membrane proteins, DcuA and DcuB (433 and 446 amino acid residues), which have 36% amino acid sequence identity. A previous amino acid sequence-based analysis predicted that DcuA and DcuB contain either 12 or 14 transmembrane helices, with the N and C termini located in the cytoplasm or periplasm (S. Six, S. C. Andrews, G. Unden, and J. R. Guest, J. Bacteriol. 176:6470-6478, 1994). These predictions were tested by constructing and analyzing 66 DcuA-BlaM fusions in which C terminally truncated forms of DcuA are fused to a beta-lactamase protein lacking the N-terminal signal peptide. The resulting topological model differs from those previously predicted. It has just 10 transmembrane helices and a central, 80-residue cytoplasmic loop between helices 5 and 6. The N and C termini are located in the periplasm and the predicted orientation is consistent with the "positive-inside rule." Two highly hydrophobic segments are not membrane spanning: one is in the cytoplasmic loop; the other is in the C-terminal periplasmic region. The topological model obtained for DcuA can be applied to DcuA homologues in other bacteria as well as to DcuB. Overproduction of DcuA to 15% of inner-membrane protein was obtained with the lacUV5-promoter-based plasmid, pYZ4. PMID- 9733686 TI - The ggpS gene from Synechocystis sp. strain PCC 6803 encoding glucosyl-glycerol phosphate synthase is involved in osmolyte synthesis. AB - A salt-sensitive mutant of Synechocystis sp. strain PCC 6803 defective in the synthesis of the compatible solute glucosylglycerol (GG) was used to search for the gene encoding GG-phosphate synthase (GGPS), the key enzyme in GG synthesis. Cloning and sequencing of the mutated region and the corresponding wild-type region revealed that a deletion of about 13 kb occurred in the genome of mutant 11. This deletion affected at least 10 open reading frames, among them regions coding for proteins showing similarities to trehalose (otsA homolog)- and glycerol-3-phosphate-synthesizing enzymes. After construction and characterization of mutants defective in these genes, it became obvious that an otsA homolog (sll1566) (T. Kaneko et al., DNA Res. 3:109-136, 1996) encodes GGPS, since only the mutant affected in sll1566 showed salt sensitivity combined with a complete absence of GG accumulation. Furthermore, the overexpression of sll1566 in Escherichia coli led to the appearance of GGPS activity in the heterologous host. The overexpressed protein did not show the salt dependence that is characteristic for the GGPS in crude protein extracts of Synechocystis. PMID- 9733685 TI - A natural large chromosomal inversion in Lactococcus lactis is mediated by homologous recombination between two insertion sequences. AB - Comparative analysis of chromosomal macrorestriction polymorphism of the two closely related Lactococcus lactis subsp. cremoris strains MG1363 and NCDO763 revealed the presence of a large inversion covering half of the genome. To determine what kind of genetic element could be implicated in this rearrangement, the two inversion junctions of MG1363 and NCDO763 chromosomes were cloned and characterized. Nucleotide sequence analysis showed the presence of one copy of the lactococcal IS905 element in each junction. Each copy of this element contained the same nucleotide mutation that inactivates the putative transposase. Comparison of the sequences surrounding the insertion sequence demonstrated that the large inversion arose from a single-step homologous recombination event between the two defective copies of the IS905 element. The large inversion presumably conferred no selective disadvantage on strain NCDO763 because this rearrangement did not alter the oriC-terC symmetry of the chromosome and the local genetic environment. PMID- 9733687 TI - Transformation of the Lyme disease spirochete Borrelia burgdorferi with heterologous DNA. AB - Studies of the spirochete Borrelia burgdorferi have been hindered by the scarcity of genetic tools that can be used in these bacteria. For the first time, a method has been developed by which heterologous DNA (DNA without a naturally occurring B. burgdorferi homolog) can be introduced into and persistently maintained by B. burgdorferi. This technique uses integration of circular DNA into the bacterial genome via a single-crossover event. The ability to transform B. burgdorferi with heterologous DNA will now permit a wide range of experiments on the biology of these bacteria and their involvement in the many facets of Lyme disease. PMID- 9733688 TI - Oxidative stress response and characterization of the oxyR-ahpC and furA-katG loci in Mycobacterium marinum. AB - Oxidative stress response in pathogenic mycobacteria is believed to be of significance for host-pathogen interactions at various stages of infection. It also plays a role in determining the intrinsic susceptibility to isoniazid in mycobacterial species. In this work, we characterized the oxyR-ahpC and furA-katG loci in the nontuberculous pathogen Mycobacterium marinum. In contrast to Mycobacterium smegmatis and like Mycobacterium tuberculosis and Mycobacterium leprae, M. marinum was shown to possess a closely linked and divergently oriented equivalents of the regulator of peroxide stress response oxyR and its subordinate gene ahpC, encoding a homolog of alkyl hydroperoxide reductase. Purified mycobacterial OxyR was found to bind to the oxyR-ahpC promoter region from M. marinum and additional mycobacterial species. Mobility shift DNA binding analyses using OxyR binding sites from several mycobacteria and a panel of in vitro generated mutants validated the proposed consensus mycobacterial recognition sequence. M. marinum AhpC levels detected by immunoblotting, were increased upon treatment with H2O2, in keeping with the presence of a functional OxyR and its binding site within the promoter region of ahpC. In contrast, OxyR did not bind to the sequences upstream of the katG structural gene, and katG expression did not follow the pattern seen with ahpC. Instead, a new open reading frame encoding a homolog of the ferric uptake regulator Fur was identified immediately upstream of katG in M. marinum. The furA-katG linkage and arrangement are ubiquitous in mycobacteria, suggesting the presence of additional regulators of oxidative stress response and potentially explaining the observed differences in ahpC and katG expression. Collectively, these findings broaden our understanding of oxidative stress response in mycobacteria. They also suggest that M. marinum will be useful as a model system for studying the role of oxidative stress response in mycobacterial physiology, intracellular survival, and other host-pathogen interactions associated with mycobacterial diseases. PMID- 9733689 TI - Mutational analysis of the transcriptional regulator GcvA: amino acids important for activation, repression, and DNA binding. AB - The GcvA protein is required for both glycine-mediated activation and purine mediated repression of the gcvTHP operon. Random and site-directed PCR mutagenesis was used to create nucleotide changes in gcvA to identify residues of the protein involved in activation, repression, and DNA binding. Single amino acid substitutions at L30 and F31 cause a defect in activation of a gcvT-lacZ fusion but have no effect on repression or DNA binding. Single amino acid substitutions at V32 and S38 cause the loss of binding of GcvA to DNA. A deletion of the carboxy-terminal 14 amino acids of GcvA results in the loss of purine mediated repression and, consequently, a constitutive activation of a gcvT-lacZ fusion. The results of this study partially define regions of GcvA involved in activation, repression, and DNA binding and demonstrate that these functions of GcvA are genetically separable. PMID- 9733690 TI - Escherichia coli tol-pal mutants form outer membrane vesicles. AB - Mutations in the tol-pal genes induce pleiotropic effects such as release of periplasmic proteins into the extracellular medium and hypersensitivity to drugs and detergents. Other outer membrane defective strains such as tolC, lpp, and rfa mutations are also altered in their outer membrane permeability. In this study, electron microscopy and Western blot analyses were used to show that strains with mutations in each of the tol-pal genes formed outer membrane vesicles after growth in standard liquid or solid media. This phenotype was not observed in tolC and rfaD cells in the same conditions. A tolA deletion in three different Escherichia coli strains was shown to lead to elevated amounts of vesicles. These results, together with plasmid complementation experiments, indicated that the formation of vesicles resulted from the defect of any of the Tol-Pal proteins. The vesicles contained outer membrane trimeric porins correctly exposed at the cell surface. Pal outer membrane lipoprotein was also immunodetected in the vesicle fraction of tol strains. The results are discussed in view of the role of the Tol-Pal transenvelope proteins in maintaining outer membrane integrity by contributing to target or integrate newly synthesized components of this structure. PMID- 9733692 TI - Efficient transfer of the pheromone-independent Enterococcus faecium plasmid pMG1 (Gmr) (65.1 kilobases) to Enterococcus strains during broth mating. AB - Plasmid pMG1 (65.1 kb) was isolated from a gentamicin-resistant Enterococcus faecium clinical isolate and was found to encode gentamicin resistance. EcoRI restriction of pMG1 produced five fragments, A through E, with molecular sizes of 50.2, 11.5, 2.0, 0.7, and 0.7 kb, respectively. The clockwise order of the fragments was ACDEB. pMG1 transferred at high frequency to Enterococcus strains in broth mating. pMG1 transferred between Enterococcus faecalis strains, between E. faecium strains, and between E. faecium and E. faecalis strains at a frequency of approximately 10(-4) per donor cell after 3 h of mating. The pMG1 transfers were not induced by the exposure of the donor cell to culture filtrates of plasmid-free E. faecalis FA2-2 or an E. faecium strain. Mating aggregates were not observed by the naked eye during broth mating. Small mating aggregates of several cells in the broth matings were observed by microscopy, while no aggregates of donor cells which had been exposed to a culture filtrate of E. faecalis FA2-2 or an E. faecium strain were observed, even by microscopy. pMG1 DNA did not show any homology in Southern hybridization with that of the pheromone-responsive plasmids and broad-host-range plasmids pAMbeta1 and pIP501. These results indicate that there is another efficient transfer system in the conjugative plasmids of Enterococcus and that this system is different from the pheromone-induced transfer system of E. faecalis plasmids. PMID- 9733691 TI - Spore photoproduct lyase from Bacillus subtilis spores is a novel iron-sulfur DNA repair enzyme which shares features with proteins such as class III anaerobic ribonucleotide reductases and pyruvate-formate lyases. AB - The major photoproduct in UV-irradiated spore DNA is the unique thymine dimer 5 thyminyl-5,6-dihydrothymine, commonly referred to as spore photoproduct (SP). An important determinant of the high UV resistance of Bacillus subtilis spores is the accurate in situ reversal of SP during spore germination by the DNA repair enzyme SP lyase. To study the molecular aspects of SP lyase-mediated SP repair, the cloned B. subtilis splB gene was engineered to encode SP lyase with a molecular tag of six histidine residues at its amino terminus. The engineered six His-tagged SP lyase expressed from the amyE locus restored UV resistance to spores of a UV-sensitive mutant B. subtilis strain carrying a deletion-insertion mutation which removed the entire splAB operon at its natural locus and was shown to repair SP in vivo during spore germination. The engineered SP lyase was purified both from dormant B. subtilis spores and from an Escherichia coli overexpression system by nickel-nitrilotriacetic acid (NTA) agarose affinity chromatography and was shown by Western blotting, UV-visible spectroscopy, and iron and acid-labile sulfide analysis to be a 41-kDa iron-sulfur (Fe-S) protein, consistent with its amino acid sequence homology to the 4Fe-4S clusters in anaerobic ribonucleotide reductases and pyruvate-formate lyases. SP lyase was capable of reversing SP from purified SP-containing DNA in an in vitro reaction either when present in a cell-free extract prepared from dormant spores or after purification on nickel-NTA agarose. SP lyase activity was dependent upon reducing conditions and addition of S-adenosylmethionine as a cofactor. PMID- 9733693 TI - Transcriptional regulation and evolution of lactose genes in the galactose lactose operon of Lactococcus lactis NCDO2054. AB - The genetics of lactose utilization within the slow-lactose-fermenting Lactococcus lactis strain NCDO2054 was studied with respect to the organization, expression, and evolution of the lac genes. Initially the beta-galactosidase gene (lacZ) was cloned by complementation of an Escherichia coli mutant on a 7-kb HpaI fragment. Nucleotide sequence analysis of the complete fragment revealed part of a gal-lac operon, and the genes were characterized by inactivation and complementation analyses and in vitro enzyme activity measurements. The gene order is galK-galT-lacA-lacZ-galE; the gal genes encode enzymes of the Leloir pathway for galactose metabolism, and lacA encodes a galactoside acetyltransferase. The galT and galE genes of L. lactis LM0230 (a lactose plasmid cured derivative of the fast-lactose-fermenting L. lactis C2) were highly similar at the nucleotide sequence level to their counterparts in strain NCDO2054 and, furthermore, had the same gene order except for the presence of the intervening lacA-lacZ strain NCDO2054. Analysis of mRNA for the gal and lac genes revealed an unusual transcriptional organization for the operon, with a surprisingly large number of transcriptional units. The regulation of the lac genes was further investigated by using fusions consisting of putative promoter fragments and the promoterless beta-glucuronidase gene (gusA) from E. coli, which identified three lactose-inducible intergenic promoters in the gal-lac operon. The greater similarity of the lacA and lacZ genes to homologs in gram-negative organisms than to those of gram-positive bacteria, in contrast to the homologies of the gal genes, suggests that the genes within the gal operon of L. lactis NCDO2054 have been recently acquired. Thus, the lacA-lacZ genes appear to have engaged the promoters of the gal operon in order to direct and control their expression. PMID- 9733694 TI - Two functionally distinct regions upstream of the cbbI operon of Rhodobacter sphaeroides regulate gene expression. AB - A number of cbbFI::lacZ translational fusion plasmids containing various lengths of sequence 5' to the form I (cbbI) Calvin-Benson-Bassham cycle operon (cbbFIcbbPIcbbAIcbbLIcbbSI) of Rhodobacter sphaeroides were constructed. Expression of beta-galactosidase was monitored under a variety of growth conditions. It was found that 103 bp of sequence upstream of the cbbFI transcription start was sufficient to confer low levels of regulated cbbI promoter expression; this activity was dependent on the presence of an intact cbbR gene. Additionally, R. sphaeroides CbbR was shown to bind to the region between 9 and 100 bp 5' to the cbbFI transcription start. Inclusion of an additional upstream sequence, from 280 to 636 bp 5' to cbbFI, resulted in a significant increase in regulated cbbI promoter expression under all growth conditions tested. A 50-bp region responsible for the majority of this increase occurs between 280 and 330 bp 5' to cbbFI. The additional 306 bp of upstream sequence from 330 to 636 bp also appears to play a positive regulatory role. A 4 bp deletion 281 to 284 bp 5' to cbbFI significantly reduced cbbI expression while the proper regulatory pattern was retained. These studies provide evidence for the presence of two functionally distinct regions of the cbbI promoter, with the distal domain providing significant regulated promoter activity that adheres to the normal pattern of expression. PMID- 9733695 TI - YscB of Yersinia pestis functions as a specific chaperone for YopN. AB - Following contact with a eucaryotic cell, Yersinia species pathogenic for humans (Y. pestis, Y. pseudotuberculosis, and Y. enterocolitica) export and translocate a distinct set of virulence proteins (YopE, YopH, YopJ, YopM, and YpkA) from the bacterium into the eucaryotic cell. During in vitro growth at 37 degrees C in the presence of calcium, Yop secretion is blocked; however, in the absence of calcium, Yop secretion is triggered. Yop secretion occurs via a plasmid-encoded type III, or "contact-dependent," secretion system. The secreted YopN (also known as LcrE), TyeA, and LcrG proteins are necessary to prevent Yop secretion in the presence of calcium and prior to contact with a eucaryotic cell. In this paper we characterize the role of the yscB gene product in the regulation of Yop secretion in Y. pestis. A yscB deletion mutant secreted YopM and V antigen both in the presence and in the absence of calcium; however, the export of YopN was specifically reduced in this strain. Complementation with a functional copy of yscB in trans completely restored the wild-type secretion phenotype for YopM, YopN, and V antigen. The YscB amino acid sequence showed significant similarities to those of SycE and SycH, the specific Yop chaperones for YopE and YopH, respectively. Protein cross-linking and immunoprecipitation studies demonstrated a specific interaction between YscB and YopN. In-frame deletions in yopN eliminating the coding region for amino acids 51 to 85 or 6 to 100 prevented the interaction of YopN with YscB. Taken together, these results indicate that YscB functions as a specific chaperone for YopN in Y. pestis. PMID- 9733696 TI - The Bacteroides fragilis BtgA mobilization protein binds to the oriT region of pBFTM10. AB - The Bacteroides fragilis conjugal plasmid pBFTM10 contains two genes, btgA and btgB, and a putative oriT region necessary for transfer in Bacteroides fragilis and Escherichia coli. The BtgA protein was predicted to contain a helix-turn helix motif, indicating possible DNA binding activity. DNA sequence analysis of the region immediately upstream of btgA revealed three sets of inverted repeats, potentially locating the oriT region. A 304-bp DNA fragment comprising this putative oriT region was cloned and confirmed to be the functional pBFTM10 oriT by bacterial conjugation experiments using E. coli and B. fragilis. btgA was cloned and overexpressed in E. coli, and the purified protein was used in electrophoretic mobility shift assays, demonstrating specific binding of BtgA protein to its cognate oriT. DNase I footprint analysis demonstrated that BtgA binds apparently in a single-stranded fashion to the oriT-containing fragment, overlapping inverted repeats I, II, and III and the putative nick site. PMID- 9733697 TI - Identification of two genes from Streptomyces argillaceus encoding glycosyltransferases involved in transfer of a disaccharide during biosynthesis of the antitumor drug mithramycin. AB - Mithramycin is an antitumor polyketide drug produced by Streptomyces argillaceus that contains two deoxysugar chains, a disaccharide consisting of two D-olivoses and a trisaccharide consisting of a D-olivose, a D-oliose, and a D-mycarose. From a cosmid clone (cosAR3) which confers resistance to mithramycin in streptomycetes, a 3-kb PstI-XhoI fragment was sequenced, and two divergent genes (mtmGI and mtmGII) were identified. Comparison of the deduced products of both genes with proteins in databases showed similarities with glycosyltransferases and glucuronosyltransferases from different sources, including several glycosyltransferases involved in sugar transfer during antibiotic biosynthesis. Both genes were independently inactivated by gene replacement, and the mutants generated (M3G1 and M3G2) did not produce mithramycin. High-performance liquid chromatography analysis of ethyl acetate extracts of culture supernatants of both mutants showed the presence of several peaks with the characteristic spectra of mithramycin biosynthetic intermediates. Four compounds were isolated from both mutants by preparative high-performance liquid chromatography, and their structures were elucidated by physicochemical methods. The structures of these compounds were identical in both mutants, and the compounds are suggested to be glycosylated intermediates of mithramycin biosynthesis with different numbers of sugar moieties attached to C-12a-O of a tetracyclic mithramycin precursor and to C-2-O of mithramycinone: three tetracyclic intermediates containing one sugar (premithramycin A1), two sugars (premithramycin A2), or three sugars (premithramycin A3) and one tricyclic intermediate containing a trisaccharide chain (premithramycin A4). It is proposed that the glycosyltransferases encoded by mtmGI and mtmGII are responsible for forming and transferring the disaccharide during mithramycin biosynthesis. From the structures of the new metabolites, a new biosynthetic sequence regarding late steps of mithramycin biosynthesis can be suggested, a sequence which includes glycosyl transfer steps prior to the final shaping of the aglycone moiety of mithramycin. PMID- 9733698 TI - Mutation of an alternative sigma factor in the cyanobacterium Nostoc punctiforme results in increased infection of its symbiotic plant partner, Anthoceros punctatus. AB - An alternative group 2 sigma factor was identified in the nitrogen-fixing, symbiotically competent cyanobacterium Nostoc punctiforme and designated sigH. Transcription of sigH was specifically induced within 1.5 h following exposure of N. punctiforme to its symbiotic plant partner, Anthoceros punctatus. A mutation in sigH resulted in a sixfold-higher initial infection of A. punctatus tissue without a parallel increase in nitrogen-fixing activity. PMID- 9733699 TI - The Na+-responsive ntp operon is indispensable for homeostatis of K+ and Na+ in Enterococcus hirae at limited proton potential. AB - Enterococcus hirae ATCC 9790 grew well in Na+-deficient, low-K+ medium, but growth was inhibited by carbonylcyanide m-chlorophenylhydrazone (CCCP). Growth inhibition and decrease of cellular K+ levels in the presence of CCCP were relieved by the addition of Na+ and a high concentration of K+. In contrast, in the mutant defective in Na+-ATPase or the NtpJ component of the KtrII K+ uptake system, CCCP-induced growth inhibition was rescued by a high concentration of K+ but not of Na+. These transporters are thus indispensable for homeostatis of K+ and Na+ at low proton potential. PMID- 9733700 TI - Transcriptional analysis and mutation of a dnaA-like gene in Synechocystis sp. strain PCC 6803. AB - Transcription of the dnaA gene of the cyanobacterium Synechocystis sp. strain PCC 6803 is light dependent and yields a monocistronic mRNA, as determined by Northern analysis. Surprisingly, mutants with inactivated dnaA were viable. In batch cultures under standard conditions, the mutants grew like the wild type and did not show an aberrant phenotype. We conclude that, unlike the situation in other bacteria, dnaA of Synechocystis sp. cannot have an essential function, such as initiation of DNA replication. PMID- 9733701 TI - Characterization of a novel acyl carrier protein, RkpF, encoded by an operon involved in capsular polysaccharide biosynthesis in Sinorhizobium meliloti. AB - Rhizobial capsular polysaccharides (RKPs) play an important role in the development of a nitrogen-fixing symbiosis with the plant host and in Sinorhizobium meliloti AK631 functional rkpABCDEF genes are required for the production of RKPs. After cloning the rkpF gene, we overexpressed and purified the derived protein product (RkpF) in Escherichia coli. Like acyl carrier protein (ACP), the RkpF protein can be labeled in vivo with radioactive beta-alanine added to the growth medium. If homogeneous RkpF protein is incubated with radiolabeled coenzyme A in the presence of purified holo-ACP synthase from E. coli, an in vitro transfer of 4'-phosphopantetheine to the RkpF protein can be observed. The conversion from apo-RkpF protein to holo-RkpF protein seems to go along with a major conformational change of the protein structure, because the holo-RkpF protein runs significantly faster on native polyacrylamide gel electrophoresis than the apo-RkpF protein. Electrospray mass spectrometric analysis reveals a mass of 9,585 for the apo-RkpF protein and a mass of 9,927 for the holo-RkpF protein. Our data show that RkpF is a novel ACP. PMID- 9733703 TI - Influence of Lif, the lysostaphin immunity factor, on acceptors of surface proteins and cell wall sorting efficiency in Staphylococcus carnosus. AB - Proteins harboring a C-terminal cell wall sorting signal are covalently linked to pentaglycine acceptors within the staphylococcal peptidoglycan. This pentaglycine was modified when the lysostaphin immunity factor (Lif) of Staphylococcus simulans was expressed in Staphylococcus carnosus, likely by the exchange of two glycine residues for serine residues. A reporter protein was efficiently linked to the modified acceptor, indicating that the sorting reaction is not strictly dependent on the wild-type structures of the acceptors. PMID- 9733704 TI - IS481 and IS1002 of Bordetella pertussis create a 6-base-pair duplication upon insertion at a consensus target site. AB - The insertion sequence IS481 and its isoform IS1002 have been observed to transpose into the bvgAS locus of Bordetella pertussis, for which the DNA sequence has previously been determined. Upon insertion of IS481 at three different sites and IS1002 at one site, a 6-bp sequence originally present was found at the junction of bvg and insertion sequence DNA. This indicates that, contrary to prior reports, IS481 and IS1002 do create a duplication upon insertion. In this light, examination of these and other examples of IS481 and IS1002 reported in the literature leads to the observation that the 6-bp recognition sequence usually fits the consensus NCTAGN. The near-palindromic nature of this sequence, when directly repeated at the ends of IS481 or IS1002, apparently led to the interpretation that 5 of these base pairs were part of the terminal inverted repeats flanking these elements. PMID- 9733702 TI - Molecular analysis of the capsule gene region of group A Streptococcus: the hasAB genes are sufficient for capsule expression. AB - Enzymes directing the biosynthesis of the group A streptococcal hyaluronic acid capsule are encoded in the hasABC gene cluster. Inactivation of hasC, encoding UDP-glucose pyrophosphorylase in the heavily encapsulated group A streptococcal strain 87-282, had no effect on capsule production, indicating that hasC is not required for hyaluronic acid synthesis and that an alternative source of UDP glucose is available for capsule production. Nucleotide sequence and deletion mutation analysis of the 5.5 kb of DNA upstream of hasA revealed that this region is not required for capsule expression. Many (10 of 23) group A streptococcal strains were found to contain insertion element IS1239' approximately 50 nucleotides upstream of the -35 site of the hasA promoter. The presence of IS1239' upstream of hasA did not prevent capsule expression. These results elucidate the molecular architecture of the group A streptococcal chromosomal region upstream of the has operon, indicate that hasABC are the sole components of the capsule gene cluster, and demonstrate that hasAB are sufficient to direct capsule synthesis in group A streptococci. PMID- 9733705 TI - Characterization of dacC, which encodes a new low-molecular-weight penicillin binding protein in Bacillus subtilis. AB - The pbp gene (renamed dacC), identified by the Bacillus subtilis genome sequencing project, encodes a putative 491-residue protein with sequence homology to low-molecular-weight penicillin-binding proteins. Use of a transcriptional dacC-lacZ fusion revealed that dacC expression (i) is initiated at the end of stationary phase; (ii) depends strongly on transcription factor sigmaH; and (iii) appears to be initiated from a promoter located immediately upstream of yoxA, a gene of unknown function located upstream of dacC on the B. subtilis chromosome. A B. subtilis dacC insertional mutant grew and sporulated identically to wild type cells, and dacC and wild-type spores had the same heat resistance, cortex structure, and germination and outgrowth kinetics. Expression of dacC in Escherichia coli showed that this gene encodes an approximately 59-kDa membrane associated penicillin-binding protein which is highly toxic when overexpressed. PMID- 9733706 TI - Cloning and molecular characterization of a multicopy, linear plasmid-carried, repeat motif-containing gene from Borrelia turicatae, a causative agent of relapsing fever. AB - Borrelia turicatae is one of several spirochete species that can cause relapsing fever. Here, we describe the identification and characterization of a gene from B. turicatae and other relapsing-fever spirochetes that exhibits homology with the rep+ and ORF-E gene families of the Lyme disease spirochetes. This gene, which we have designated repA, encodes a putative protein of 30.2 kDa with an isoelectric point of 4.69. The central region of RepA harbors a series of amino acid repeat motifs which exhibit homology with casein kinase 2 phosphorylation sites. Through Southern hybridization analyses, we demonstrate that repA (or a closely related sequence) is multicopy in the relapsing-fever spirochetes and is carried on variably sized linear plasmids in both Borrelia parkeri and B. turicatae. Transcriptional analyses demonstrate that repA is expressed, albeit at low levels, during in vitro cultivation of B. turicatae. Transcriptional start site analysis revealed that repA is preceded by a consensus ribosomal binding site and an appropriately spaced promoter element. The sequence conservation, unique features, and multicopy status of repA and its homologs suggest that RepA may play an important genus-wide role in the biology of the Borrelia. PMID- 9733707 TI - Heterologous expression of the Desulfovibrio gigas [NiFe] hydrogenase in Desulfovibrio fructosovorans MR400. AB - The ability of Desulfovibrio fructosovorans MR400 DeltahynABC to express the heterologous cloned [NiFe] hydrogenase of Desulfovibrio gigas was investigated. The [NiFe] hydrogenase operon from D. gigas, hynABCD, was cloned, sequenced, and introduced into D. fructosovorans MR400. A portion of the recombinant heterologous [NiFe] hydrogenase was totally matured, exhibiting catalytic and spectroscopic properties identical to those of the native D. gigas protein. A chimeric operon containing hynAB from D. gigas and hynC from D. fructosovorans placed under the control of the D. fructosovorans hynAp promoter was constructed and expressed in D. fructosovorans MR400. Under these conditions, the same level of activity was obtained as with the D. gigas hydrogenase operon. PMID- 9733708 TI - A region in Bacillus subtilis sigmaH required for Spo0A-dependent promoter activity. AB - Spo0A activates transcription in Bacillus subtilis from promoters that are used by two types of RNA polymerase, RNA polymerase containing the primary sigma factor, sigmaA, and RNA polymerase containing a secondary sigma factor, known as sigmaH. The region of sigmaA near positions 356 to 359 is required for Spo0A dependent promoter activation, possibly because Spo0A interacts with this region of sigmaA at these promoters. To determine if the amino acids in the corresponding region of sigmaH are also important in Spo0A-dependent promoter activation, we examined the effects of single alanine substitutions at 10 positions in sigmaH (201 to 210). Two alanine substitutions in sigmaH, at glutamine 201 (Q201A) and at arginine 205 (R205A), significantly decreased activity from the Spo0A-dependent, sigmaH-dependent promoter spoIIA but did not affect expression from the sigmaH-dependent, Spo0A-independent promoters citGp2 and spoVG. Therefore, promoter activation by Spo0A requires homologous regions in sigmaA and sigmaH. A mutant form of Spo0A, S231F, that suppresses the sporulation defect caused by several amino acid substitutions in sigmaA did not suppress the sporulation defects caused by the Q201A and R205A substitutions in sigmaH. This result and others indicate that different surfaces of Spo0A probably interact with sigmaA and sigmaH RNA polymerases. PMID- 9733709 TI - Mammalian lipid phosphate phosphohydrolases. PMID- 9733710 TI - Requirement of p38 mitogen-activated protein kinase for neuronal differentiation in PC12 cells. AB - Nerve growth factor (NGF) induces sustained activation of classical MAP kinase (MAPK, also known as ERK) and neuronal differentiation in PC12 cells, whereas epidermal growth factor (EGF) induces transient activation of ERK/MAPK and stimulates proliferation of the cells. Although previous studies showed that sustained activation of ERK/MAPK is important for neuronal differentiation of the cells, a recent report revealed that inhibition of the sustained phase of ERK/MAPK activation alone does not block neurite outgrowth caused by NGF. These results suggest requirement for an additional signaling pathway(s) triggered by NGF in neuronal differentiation. Here we show that NGF induces sustained activation of p38, a subfamily member of the MAPK superfamily, and that inhibition of the p38 pathway blocks neurite outgrowth in PC12 cells. Surprisingly, expression of constitutively active MAPK/ERK kinase (MAPKK, also known as MEK) results in p38 activation as well as ERK/MAPK activation, and a p38 inhibitor blocks neurite outgrowth caused by the constitutively active MAPKK/MEK. Moreover, constitutive activation of p38 is able to induce neurite outgrowth when combined with EGF treatment. These results reveal an essential role of p38 in neuronal differentiation in PC12 cells. PMID- 9733711 TI - Cloning and expression analysis of a novel salicylate suppressible gene, Hs-CUL 3, a member of cullin/Cdc53 family. AB - By using a mRNA differential display technique to search for salicylate suppressible genes, we identified a cDNA in human foreskin fibroblasts, which by GenBankTM DNA data base search shows sequence homology to the recently reported cullin/Cdc53 (CUL) family genes, especially CUL-3. We have cloned the full-length human CUL-3 (Hs-CUL-3) cDNA. It encodes a 768-amino acid polypeptide and has a predicted molecular weight of 88,939. The amino acid sequence of Hs-CUL-3 shows 46% homology to that of its Caenorhabditis elegans ortholog, Ce-CUL-3, and 27 and 23% to that of Hs-CUL-1 and Hs-CUL-2, respectively. Northern blot analysis showed that phorbol 12-myristate 13-acetate increased the expression of Hs-CUL-3 mRNA in a concentration- and time-dependent manner, and this increase was inhibited by sodium salicylate. Hs-CUL-3 widely expressed in human tissues and its expression in cultured COLO205 colon cancer cells was increased when compared with that in normal colon cells. It is likely that Hs-CUL-3 is involved in cell proliferation control. PMID- 9733712 TI - Smad7 is an activin-inducible inhibitor of activin-induced growth arrest and apoptosis in mouse B cells. AB - Members of the transforming growth factor-beta (TGF-beta) family, which includes the activins, relay signals from serine/threonine kinase receptors in membrane to nucleus via intracellular Sma- and Mad-related (Smad) proteins. Inhibitory Smad proteins were found to prevent the interaction between the serine/threonine kinase receptors and pathway-restricted Smad proteins. Smad7 was identified as a TGF-beta-inducible antagonist of TGF-beta signaling, and it may participate in a negative feedback loop to control TGF-beta signaling. Here we demonstrate that the mRNA expression of Smad7 is induced by activin A in mouse B cell hybridoma HS 72 cells, which undergo growth arrest and apoptosis upon exposure to activin A. The ectopic expression of mouse Smad7 in HS-72 cells suppressed the activin A induced cell cycle arrest in the G1 phase by abolishing the activin A-induced expression of p21(CIP1/WAF1) and hypophosphorylation of retinoblastoma protein. Furthermore, Smad7 expression suppressed activin A-induced apoptosis in HS-72 cells. Thus, our data indicate that Smad7 is an activin A-inducible antagonist of activin A-induced growth arrest and apoptosis of B lineage cells. PMID- 9733713 TI - Activity of Rap1 is regulated by bombesin, cell adhesion, and cell density in NIH3T3 fibroblasts. AB - Rap1 and Ras are homologous GTPases that are implicated in cell proliferation and differentiation. At present, little is known about the regulation of Rap1 activity. Using a recently developed assay with activation-specific probes, we found increased activity of endogenous Rap1 in NIH3T3 cells after stimulation with the neuropeptide growth factor bombesin in a concentration- and time dependent manner. The activity of endogenous Ras was unaffected. Analysis of putative effectors showed no activation of c-Raf-1 or B-Raf after bombesin stimulation. However, MAPK/Erk-phosphorylation and the proliferation rate was increased. In addition, Rap1 was activated during cell adhesion to coated and uncoated tissue culture plates, as well as in response to various mitogens. Surprisingly, the basal Rap1 activity was observed to be cell density-dependent, with low levels when cells were reaching confluency. The results suggest that Rap1 acts as an important mediator of mitogenic signals distinct to Ras activation. PMID- 9733715 TI - Transpeptidation by porcine pepsin catalyzed by a noncovalent intermediate unique to its iso-mechanism. AB - Porcine pepsin proteolysis of the hexapeptide Leu-Ser-p-nitro-Phe-Nle-Ala-Leu-OMe (where OMe = methoxy and Nle = norleucine) in the presence of dipeptide Leu-Leu synthesizes a new hexapeptide Leu-Ser-p-nitro-Phe-Leu-Leu. Contrary to transpeptidation kinetics of other proteases, which depend upon an acyl-enzyme intermediate, the time course for pepsin-catalyzed transpeptidation displays a distinct lag before reaching a steady-state reaction velocity. Moreover, this lag is coupled to burst kinetics for the formation of proteolytic products, Leu-Ser-p nitro-Phe and Nle-Ala-Leu-OMe. The lag requires that free Leu-Ser-p-nitro-Phe accumulate in the reaction medium during the lag phase and subsequently rebind for transpeptidation. Consistent with this dissociative kinetic mechanism are normal solvent isotope effects on formation of the proteolytic products Leu-Ser-p nitro-Phe (vH/vD = 2.2 +/- 0.2) and Nle-Ala-Leu-OMe (vH/vD = 1.8 +/- 0.1) as opposed to an inverse effect on the formation of the transpeptidation product Leu Ser-p-nitro-Phe-Leu-Leu (vH/vD = 0.40 +/- 0.09). Because proteolysis is slower in D2O but transpeptidation is faster, the isotopically sensitive step must occur after release of both products of proteolysis, which precludes putative acyl enzyme covalent intermediates. Isotopically enhanced transpeptidation is a new type of isotope effect but one that is consistent with the Uni Bi iso-mechanism previously postulated on the basis of solvent isotope effects on Vmax but not on Vmax/Km (Rebholz, K. L., and Northrop, D. B. (1991) Biochem. Biophys Res. Commun. 179, 65-69) and confirmed by solvent isotope effects on the onset of inhibition by pepstatin (Cho, Y.-K., Rebholz, K. L., and Northrop, D. B. (1994) Biochemistry 33, 9637-9642). As a new biochemical mechanism for peptide bond synthesis that has a potential for applications in biotechnology, it is here proposed that the energy necessary to drive peptide synthesis from free peptides comes from the sizable free energy drop associated with rehydration of the active site of pepsin in 55 M water. PMID- 9733714 TI - Grb2 interaction with MEK-kinase 1 is involved in regulation of Jun-kinase activities in response to epidermal growth factor. AB - Epidermal growth factor (EGF) receptor was shown to be involved in the activation pathway of the stress-activated protein kinase/c-Jun NH2-terminal kinase (SAPK/JNK) cascade not only by EGF, but also by UV radiation or osmotic stress. This paper describes a specific interaction between the COOH-terminal SH3 domain of Grb2 and the NH2-terminal regulatory domain of MEKK1 in ER22 cells overexpressing the EGF receptor. This interaction results in the formation of a constitutive complex between Grb2 and MEKK1 in both proliferating and resting cells. EGF stimulation causes this complex to be rapidly and transiently recruited by Shc proteins. The subsequent release of the Grb2-MEKK1 complex from Shc proteins correlates with JNK activation. Transfection of the NH2-terminal regulatory domain of MEKK1 specifically inhibits EGF-dependent JNK activation indicating that Grb2 is involved in MEKK1 activation. Thus, adaptor proteins have a new role in the regulation of the SAPK/JNK cascade after EGF stimulation. PMID- 9733716 TI - Phosphatidylinositides bind to plasma membrane CD14 and can prevent monocyte activation by bacterial lipopolysaccharide. AB - Although bacterial lipopolysaccharides (LPS) and several other microbial agonists can bind to mCD14 (membrane CD14), a cell-surface receptor found principally on monocytes and neutrophils, host-derived mCD14 ligands are poorly defined. We report here that phosphatidylinositol (PtdIns), phosphatidylinositol-4-phosphate, and other phosphatidylinositides can bind to mCD14. Phosphatidylserine (PS), another anionic glycerophospholipid, binds to mCD14 with lower apparent affinity than does PtdIns. LPS-binding protein, a lipid transfer protein found in serum, facilitates both PS- and PtdIns-mCD14 binding. PtdIns binding to mCD14 can be blocked by anti-CD14 monoclonal antibodies that inhibit LPS-mCD14 binding, and PtdIns can inhibit both LPS-mCD14 binding and LPS-induced responses in monocytes. Serum-equilibrated PtdIns also binds to mCD14-expressing cells, raising the possibility that endogenous PtdIns may modulate cellular responses to LPS and other mCD14 ligands in vivo. PMID- 9733717 TI - Phosphatidylinositol 3,4,5-trisphosphate-dependent stimulation of phospholipase C gamma2 is an early key event in FcgammaRIIA-mediated activation of human platelets. AB - Platelets express a single class of Fcgamma receptor (FcgammaRIIA), which is involved in heparin-associated thrombocytopenia and possibly in inflammation. FcgammaRIIA cross-linking induces platelet secretion and aggregation, together with a number of cellular events such as tyrosine phosphorylation, activation of phospholipase C-gamma2 (PLC-gamma2), and calcium signaling. Here, we show that in response to FcgammaRIIA cross-linking, phosphatidylinositol (3,4, 5) trisphosphate (PtdIns(3,4,5)P3) is rapidly produced, whereas phosphatidylinositol (3,4)-bisphosphate accumulates more slowly, demonstrating a marked activation of phosphoinositide 3-kinase (PI 3-kinase). Inhibition of PI 3-kinase by wortmannin or LY294002 abolished platelet secretion and aggregation, as well as phospholipase C (PLC) activation, indicating a role of this lipid kinase in the early phase of platelet activation. Inhibition of PLCgamma2 was not related to its tyrosine phosphorylation state, since wortmannin actually suppressed its dephosphorylation, which requires platelet aggregation and integrin alphaIIb/beta3 engagement. In contrast, the stable association of PLCgamma2 to the membrane/cytoskeleton interface observed at early stage of platelet activation was fully abolished upon inhibition of PI 3-kinase. In addition, PLCgamma2 was able to preferentially interact in vitro with PtdIns(3,4,5)P3. Finally, exogenous PtdIns(3,4,5)P3 restored PLC activation in permeabilized platelets treated with wortmannin. We propose that PI 3-kinase and its product PtdIns(3,4,5)P3 play a key role in the activation and adequate location of PLCgamma2 induced by FcgammaRIIA cross-linking. PMID- 9733718 TI - The second intracellular loop of the m5 muscarinic receptor is the switch which enables G-protein coupling. AB - We have completed a systematic search of the intracellular loops of a muscarinic acetylcholine receptor for domains that govern G-protein coupling. A unique feature of the second intracellular (i2) loop was an ordered cluster of residues where diverse substitutions cause constitutive activation. A second group of residues in i2 was identified where mutations compromised receptor/G-protein coupling. The residues of each group alternate and are spaced three to four positions apart, suggesting an alpha-helical structure where these groups form opposing faces of the helix. We propose that the constitutively activating face normally constrains the receptor in the "off-state," while the other face couples G-proteins in the "on-state." Therefore, the i2 loop functions as the switch enabling G-protein activation. PMID- 9733719 TI - Selective interference of beta-arrestin 1 with kappa and delta but not mu opioid receptor/G protein coupling. AB - The role of beta-arrestin 1 (beta-arr1) in regulation of responsiveness of kappa, delta, and mu opioid receptors has been investigated in human embryonic kidney 293 cells cotransfected with opioid receptor and beta-arr1. Expression of human beta-arr1 attenuated kappa and delta opioid receptor subtype-mediated inhibition of cAMP production and resulted in a 100-fold increase of EC50 values for kappa agonist U69593 and delta-agonist [D-Pen2, D-Pen5]enkephalin and 30-40% reduction of their maximal responses. In contrast, coexpression of beta-arr1 with mu opioid receptor did not affect the concentration-effect relationship of mu-agonist [D Ala2,N-Me-Phe4,Gly5-ol]enkephalin. In parallel, kappa and delta receptor-mediated G protein activation was also remarkably attenuated by overexpression of beta arr1, while the mu-agonist-stimulated response remained intact. These results indicate that beta-arr1 interferes receptor/G protein coupling and differentially regulates the responsiveness of opioid receptors. Truncation of kappa and delta opioid receptors at carboxyl termini abolished inhibition of beta-arr1 on the responsiveness of both receptors. Furthermore, mu opioid receptor became sensitive to beta-arr1 regulation following replacement of its carboxyl terminus with the corresponding portion of the delta receptor. Removal of potential phosphorylation sites on the carboxyl terminus of kappa opioid receptor led to reduced effect of beta-arr1 on the receptor-mediated response. These results suggest that receptor carboxyl terminus and its phosphorylation play an important role in the interaction of beta-arr1 and opioid receptors. PMID- 9733720 TI - Half-site modification of Lys-480 of the Na+,K+-ATPase alpha-chain with pyridoxal 5'-diphospho-5'-adenosine reduces ATP-dependent phosphorylation stoichiometry from half to a quarter. AB - Pig and dog kidney Na+,K+-ATPase preparations, irrespective of specific activity, showed approximately 0.5 mol of maximum phosphorylation/mol alpha-chain for ATP or acetyl phosphate (AcP) at steady state conditions. Pyridoxal 5'-diphospho-5' adenosine (AP2PL)-treated pig kidney enzymes containing approximately 0.5 mol of AP2PL probe at Lys-480/mol (Tsuda, T., Kaya, S., Funatsu, H., Hayashi, Y., and Taniguchi, K. (1998) J. Biochem. (Tokyo) 123, 169-174) showed a quarter-site phosphorylation by ATP and half-site phosphorylation from AcP. The addition of 10 microM ATP to the Mg2+-Na+-bound AP2PL enzyme induced rapid quarter-site phosphorylation (47/s), followed by two different AP2PL fluorescence changes, a rapid decrease (29/s) and a slow increase (1.1/s). The addition of 1 mM AcP to the Mg2+-Na+-bound AP2PL enzyme induced a slow half-site phosphorylation (3/s), followed by a monophasic AP2PL fluorescence increase (1.2/s). After treatment of the AP2PL enzyme with fluorescein 5'-isothiocyanate to modify Lys-501 fully, the Mg2+-Na+-dependent phosphorylation capacity from ATP of the resulting AP2PL fluorescein 5'-isothiocyanate enzyme was reduced to approximately 6% without significant changes in half-site phosphorylation capacity with respect to AcP, dynamic AP2PL fluorescence change by ATP and change by AcP. These data and others support the hypothesis that the functional membrane-bound Na+, K+-ATPase has tetrameric properties. PMID- 9733721 TI - ATP and acetyl phosphate induces molecular events near the ATP binding site and the membrane domain of Na+,K+-ATPase. The tetrameric nature of the enzyme. AB - The addition of ATP to Mg2+-Na+-bound-probe labeled Na+,K+-ATPase preparations containing approximately 0.5 mol of pyridoxal 5'-diphospho-5'-adenosine (AP2PL) probe at Lys-480 and approximately 0.9 mol of fluorescein 5'-isothiocyanate (FITC) probe at Lys-501 showed a decrease and an increase in the AP2PL fluorescence intensity with neither significant ATP-dependent phosphorylation nor FITC fluorescence change. The rate constants for the fluorescence change increased nearly linearly with increasing ATP concentrations. The substitution of AcP for ATP decreased the FITC fluorescence rather monophasically, 8.5/s, which was followed by the half-site phosphorylation with same amount of components with different rate constant, 7.2 and 4.6/s, followed by a much slower increase in the two components of AP2PL fluorescence, 1.4 and 0.2/s. The addition of Na+ with increasing concentrations of ATP to the K+-bound AP2PL-FITC enzymes induced accelerations in the decrease and an increase in the AP2PL fluorescence intensity with two different increases in the FITC fluorescence intensity, showing that the same concentration of ATP is capable of inducing four different fluorescence changes. The addition of ATP to the Mg2+-Na+-bound enzymes modified with N-[p-(2 benzimidazolyl)phenyl]-maleimide (BIPM) at Cys-964 and retaining full Na+,K+ ATPase activity induced two different increases in BIPM fluorescence intensity. Each rate constant for the BIPM fluorescence change versus concentrations of ATP gave two intersecting straight lines. These data and the stoichiometries of fluorescence probe bindings and ATP- and AcP-dependent phosphorylation provide strong support for the conclusion that the functional membrane-bound Na+,K+ ATPase is a tetramer. PMID- 9733722 TI - Signaling and phosphorylation-impaired mutants of the rat follitropin receptor reveal an activation- and phosphorylation-independent but arrestin-dependent pathway for internalization. AB - We have previously shown that the rat follitropin receptor (rFSHR) expressed in transfected cells becomes phosphorylated upon stimulation of the cells with agonist or a phorbol ester. Peptide mapping and mutagenesis studies have also shown that the agonist- or phorbol ester-induced phosphorylation of the rFSHR maps to Ser/Thr residues present in the first and third intracellular loops. The experiments presented herein were initially designed to test for the presence of additional phosphorylation sites on the second intracellular loop of the rFSHR. Analysis of two new mutants in which the two threonines in the second intracellular loop (rFSHR-2L) or the two threonines in the second intracellular loop and the seven Ser/Thr residues in the third intracellular loop (rFSHR-2L + 3L) were mutated showed that one or more of the two threonines in the second intracellular loop are phosphorylated in response to phorbol ester, but not in response to agonist stimulation. Since rFSHR-2L and rFSHR-2L + 3L displayed a reduction in agonist-induced signaling, two additional mutants (rFSHR-D389N and rFSHR-Y530F) were constructed in an attempt to better understand the relationship between the agonist-induced activation, phosphorylation, and internalization of the rFSHR. These point mutations impaired agonist-stimulated signal transduction and abolished agonist-induced phosphorylation. Co-transfection studies revealed that the phosphorylation of these mutants can be rescued by overexpression of G protein-coupled receptor kinase 2, but this increased phosphorylation only rescues the internalization of rFSHR-D389N. The internalization of both mutants could be rescued by overexpression of arrestin-3, however. Taken together, these results argue that the agonist-induced activation and phosphorylation of the rFSHR are not essential for internalization. while the interaction of the rFSHR with a nonvisual arrestin is essential for internalization. PMID- 9733724 TI - Plasma membrane-bound tissue inhibitor of metalloproteinases (TIMP)-2 specifically inhibits matrix metalloproteinase 2 (gelatinase A) activated on the cell surface. AB - The cell-surface activation of pro-matrix metalloproteinase 2 (pro-MMP-2) is considered to be critical for cell migration and invasion. Treatment of human uterine cervical fibroblasts with concanavalin A activates pro-MMP-2 on the cell surface by converting it to the 65-kDa form with a minor form of 45 kDa. However, the 65-kDa MMP-2 was inactivated by tissue inhibitor of metalloproteinases (TIMP) 2 that was bound to the plasma membrane upon concanavalin A treatment. TIMP-2 binds to the plasma membrane through its N-terminal domain by two different modes of interaction as follows: one is sensitive to a hydroxamate (HXM) inhibitor of MMPs and the other is HXM-insensitive. TIMP-2 bound to the membrane in a HXM insensitive manner, comprising about 40-50% of TIMP-2 on the membrane, is the inhibitor of the cell surface-activated MMP-2. It, however, does not inhibit MMP 3, MMP-9, and the 45-kDa MMP-2 lacking the C-terminal domain. The inhibition of the 65-kDa MMP-2 by TIMP-2 is initiated by the interaction of their C-terminal domains. Subsequently, the MMP-2.TIMP-2 complex is released from the membrane, and the activity of MMP-2 is blocked by TIMP-2. In the presence of collagen types I, II, III, V, or gelatin, the rate of inhibition of the 65-kDa MMP-2 by the membrane-bound TIMP-2 decreased considerably. These results suggest that the pericellular activity of MMP-2 is tightly regulated by membrane-bound TIMP-2 and surrounding extracellular matrix components. PMID- 9733723 TI - Increased association of synaptosome-associated protein of 25 kDa with syntaxin and vesicle-associated membrane protein following acrosomal exocytosis of sea urchin sperm. AB - Synaptosomal-associated protein of 25 kDa (SNAP-25) is a palmitoylated integral membrane protein expressed almost exclusively in neuronal and neuroendocrine tissues. This protein forms a ternary complex with vesicle-associated membrane protein (VAMP) and syntaxin, which is thought to regulate the fusion of plasma and vesicle membranes during exocytosis. We report the identification of SNAP-25 expressed in sea urchin sperm. Sea urchin SNAP-25 shares greater identity with mammalian SNAP-25 than with mammalian SNAP-23, a ubiquitously expressed homologue believed to regulate membrane fusion in non-neuronal tissues. Sea urchin sperm contain a single exocytotic vesicle, the acrosomal vesicle, whose contents are exposed during the acrosome reaction. Fusion of the plasma membrane with the acrosomal vesicle membrane at multiple points (vesiculation) results in the release of SNAP-25 with the shed acrosome reaction vesicles. A complex containing SNAP-25, syntaxin, and VAMP is present in sperm, as detected by affinity chromatography and immunoprecipitation. Although this complex is present prior to the acrosome reaction, the amount of complex increases over 4-fold following acrosomal exocytosis. These findings support the involvement of SNAP-25 in the invertebrate sperm acrosome reaction, possibly through increased association with VAMP and syntaxin driving the fusion of plasma and acrosomal membranes. PMID- 9733726 TI - Inhibition of ribonuclease P activity by retinoids. AB - The effect of two naturally occurring (retinol and all-trans retinoic acid) and two synthetic (isotretinoin and acitretin) analogs of vitamin A (retinoids) on tRNA biogenesis was investigated employing the RNase P of Dictyostelium discoideum as an in vitro experimental system. RNase P is an ubiquitous and essential enzyme that endonucleolytically cleaves all tRNA precursors to produce the mature 5' end. All retinoids tested revealed a dose-dependent inhibition of RNase P activity, indicating that these compounds may have a direct effect on tRNA biogenesis. Detailed kinetic analysis showed that all retinoids behave as classical competitive inhibitors. The Ki values determined were 1475 microM for retinol, 15 microM for all-trans retinoic acid, 20 microM for isotretinoin, and 8.0 microM for acitretin. On the basis of these values acitretin is a 184, 2.5, and 1.9 times more potent inhibitor, as compared with retinol, isotretinoin, and all-trans retinoic acid, respectively. Taking into account that retinoids share no structural similarities to precursor tRNA, it is suggested that their kinetic behavior reflects allosteric interactions of these compounds with hydrophobic site(s) of D. discoideum RNase P. PMID- 9733725 TI - In the uncoupling protein (UCP-1) His-214 is involved in the regulation of purine nucleoside triphosphate but not diphosphate binding. AB - The nucleotide binding to uncoupling protein (UCP-1) of brown adipose tissue is regulated by pH. The binding pocket of the nucleotide phosphate moiety has been proposed to be controlled by the protonization of a carboxyl group (pK approximately 4.5) for both nucleoside diphosphates (NDP) and nucleoside triphosphates (NTP) (identified as Glu-190) and of a histidine (pK approximately 7. 2) for NTP only. Here we identify His-214 as a pH sensor specific for NTP binding only. In reconstituted UCP-1 from hamster, DEPC diminishes binding of NTP but not of NDP. It also prevents inhibition of H+ transport by NTP but not by NDP. Hamster UCP-1 expressed in Saccharomyces cerevisiae was mutated to H214N resulting in only moderate change of the binding affinity for NTP (GTP) but a 10 fold affinity decrease with the bulkier substituent in H214W, whereas the affinity for NDP (ADP) was largely unchanged. The steep decrease with pH of the binding affinity for NTP in wild type (from pH 6.0 to 7.5) was much flatter in the mutants. Also, the pH dependence of binding and dissociation rates was diminished in these mutants. The transport of H+ and Cl- was not affected. Thus, His-214 is only involved in nucleotide binding, whereas, as previously shown, His 145 and His-147 are involved only in H+ transport. The results validate the earlier proposal of a histidine regulating the NTP binding in addition to a carboxyl group controlling both NTP and NDP binding. It is proposed that His-214 protrudes into the binding pocket for the gamma-phosphate thus inhibiting NTP binding and that His214H+ is retracted by a background -CO2- group to give way for the gamma-phosphate. PMID- 9733727 TI - Purification, cloning, and preliminary characterization of a Spiroplasma citri ribosomal protein with DNA binding capacity. AB - The rpsB-tsf-x operon of Spiroplasma citri encodes ribosomal protein S2 and elongation factor Ts, two components of the translational apparatus, and an unidentified X protein. A potential DNA-binding site (a 20-base pair (bp) inverted repeat sequence) is located at the 3' end of rpsB. Southwestern analysis of S. citri proteins, with a 30-bp double-stranded oligonucleotide probe (IRS), containing the 20-bp inverted repeat sequence and the genomic flanking sequences, detected an IRS-binding protein of 46 kDa (P46). P46 protein, which displays preferential affinity for the IRS, was purified from S. citri by a combination of affinity and gel filtration chromatographies. The native form of P46 seems to be homomultimeric as estimated by SDS-polyacrylamide gel electrophoresis analysis and gel filtration. A 3.5-kilobase pair S. citri DNA fragment comprising the P46 gene and flanking sequences was cloned and sequenced. Sequence analysis of this DNA fragment indicated that the P46 gene is located within the S10-spc operon of S. citri at the position of the gene coding for ribosomal protein L29 in the known S10-spc operons. The similarity between the N-terminal domain of P46 and the L29 ribosomal protein family and the presence of a 46-kDa IRS-binding protein in S. citri ribosomes indicated that P46 is the L29 ribosomal protein of S. citri. We suggest that P46 is a bifunctional protein with an L29 N-terminal domain and a C-terminal domain involved in IRS binding. PMID- 9733728 TI - Regulation of human involucrin promoter activity by a protein kinase C, Ras, MEKK1, MEK3, p38/RK, AP1 signal transduction pathway. AB - Involucrin is a marker of keratinocyte terminal differentiation. Our previous studies show that involucrin mRNA levels are increased by the keratinocyte differentiating agent, 12-O-tetradecanoylphorbol-13-acetate (TPA) (Welter, J. F., Crish, J. F., Agarwal, C., and Eckert, R. L. (1995) J. Biol. Chem. 270, 12614 12622). We now study the signaling cascade responsible for this regulation. Protein kinase C and tyrosine kinase inhibitors inhibit both the TPA-dependent mRNA increase and the TPA-dependent increase in hINV promoter activity. The relevant response element is located within the promoter proximal regulatory region and includes an AP1 site, AP1-1. Co-transfection of the hINV promoter with dominant negative forms of Ras, MEKK1, MEK1, MEK7, MEK3, p38/RK, and c-Jun inhibit the TPA-dependent increase. Wild type MEKK1 enhances promoter activity and the activity can be inhibited by dominant negative MEKK1, MEK1, MEK7, MEK3, p38/RK, and c-Jun. In contrast, wild type Raf-1, ERK1, ERK2, MEK4, or JNK1 produced no change in activity and the dominant negative forms of these kinases failed to suppress TPA-dependent transcription. Treatment with an S6 kinase (S6K) inhibitor, or transfection with constitutively active S6K produced relatively minor changes in promoter activity, ruling out a regulatory role for S6K. These results suggest that activation of involucrin transcription involves a pathway that includes protein kinase C, Ras, MEKK1, MEK3, and p38/RK. Additional pathways that transfer MEKK1 activation via MEK1 and MEK7 also may function, but the downstream targets of these kinases need to be identified. AP1 transcription factors appear to be the ultimate target of this regulation. PMID- 9733729 TI - Rapid identification of protein phosphatase 1-binding proteins by mixed peptide sequencing and data base searching. Characterization of a novel holoenzymic form of protein phosphatase 1. AB - Microcystin-affinity chromatography was used to purify 15 protein phosphatase 1 (PP1)-binding proteins from the myofibrillar fraction of rabbit skeletal muscle. To reduce the time and amount of material required to identify these proteins, proteome analysis by mixed peptide sequencing was developed. Proteins are resolved by SDS-polyacrylamide gel electrophoresis, electroblotted to polyvinylidene fluoride membrane, and stained. Bands are sliced from the membrane, cleaved briefly with CnBr, and applied without further purification to an automated Edman sequencer. The mixed peptide sequences generated are sorted and matched against the GenBank using two new programs, FASTF and TFASTF. This technology offers a simple alternative to mass spectrometry for the subpicomolar identification of proteins in polyacrylamide gels. Using this technology, all 15 proteins recovered in PP-1C affinity chromatography were sequenced. One of the proteins, PP-1bp55, was homologous to human myosin phosphatase, MYPT2. A second, PP-1bp80, identified in the EST data bases, contained a putative PP-1C binding site and a nucleotide binding motif. Further affinity purification over ATP Sepharose isolated PP-1bp80 in a quaternary complex with PP-1C and two other proteins, PP-1bp29 and human p20. Recombinant PP-1bp80 also bound PP-1C and suppressed its activity toward a variety of substrates, suggesting that the protein is a novel regulatory subunit of PP-1. PMID- 9733731 TI - The Gcn5.Ada complex potentiates the histone acetyltransferase activity of Gcn5. AB - The Gcn5 histone acetyltransferase (HAT) is part of a large multimeric complex that is required for transcriptional activation in yeast. This complex can acetylate in vitro and in a Gcn5-dependent manner both nucleosomal and free core histones. For this reason it is believed that part of the function of the Gcn5.Ada complex is chromatin remodeling effected by histone acetylation. The roles of the other subunits of this complex are not yet known. We have generated mutated Gcn5 proteins with severely attenuated in vitro HAT activities. Despite their apparent loss in HAT activity, these GCN5 derivatives complemented all the defects of a gcn5 strain. We have shown that when these mutated proteins were produced in yeast cells in the absence of another component of the complex, Ada2, their activity was still compromised. By contrast, when produced in the wild type context, they were partially capable of acetylating free histones and were even more active when nucleosomal arrays were used as substrates. Kinetic enzymatic analyses showed that the rate of catalysis by Gcn5 was enhanced when the mutated proteins were produced in yeast in the presence of Ada2. Because Ada2 is required for the assembly of Gcn5, we conclude that one role for components of the Gcn5.Ada complex is the potentiation of its HAT activity. PMID- 9733730 TI - The topology of VDAC as probed by biotin modification. AB - The outer membrane of mitochondria contains channels called VDAC (mitochondrial porin), which are formed by a single 30-kDa protein. Cysteine residues introduced by site-directed mutagenesis at sites throughout Neurospora crassa VDAC (naturally devoid of cysteine) were specifically biotinylated prior to reconstitution into planar phospholipid membranes. From previous studies, binding of streptavidin to single biotinylated sites results in one of two effects: reduced single-channel conductance without blockage of voltage gating (type 1) or locking of the channels in a closed conformation (type 2). All sites react with streptavidin only from one side of the membrane. Here, we extend this approach to VDAC molecules containing two cysteines and determine the location of each biotinylated residue with respect to the other within the membrane. When a combination of a type 1 and a type 2 site was used, each site could be observed to react with streptavidin. Two sets of sites located on opposite surfaces of the membrane were identified, thereby establishing the transmembrane topology of VDAC. A revised folding pattern for VDAC, consisting of 1 alpha helix and 13 beta strands, is proposed by combining these results with previously obtained information on which sites are lining the aqueous pore. PMID- 9733732 TI - Kringle 2 mediates high affinity binding of plasminogen to an internal sequence in streptococcal surface protein PAM. AB - Many cells express receptors for plasminogen (Pg), although the responsible molecules in most cases are poorly defined. In contrast, the group A streptococcal surface protein PAM contains a domain with two 13-amino acid residue long repeated sequences (a1 and a2) responsible for Pg binding. Here we identify the region in Pg that interacts with PAM. A radiolabeled proteolytic plasminogen fragment containing the first three kringles (K1-K3) interacted with streptococci expressing PAM or a chimeric surface protein harboring the a1a2 sequence. In contrast, plasminogen fragments containing kringle 4 or kringle 5 and the activable serine proteinase domain failed to bind to PAM-expressing group A streptococci. A synthetic and a recombinant polypeptide containing the a1a2 sequence both bound to immobilized recombinant K2 (rK2) but not to rK1 or rK3. The interaction between the a repeat region and rK2 was reversible, and rK2 completely blocked the binding of Pg to the a1a2 region. The binding of the a repeat containing polypeptide to K2 occurred with an equilibrium association constant of 4.5 x 10(7) M-1, as determined by surface plasmon resonance, a value close to that (1.6 x 10(7) M-1) calculated for the a1a2-Pg interaction. Inhibition experiments suggested involvement of the lysine-binding site of K2 in the interaction. These data demonstrate that K2 contains the major Pg-binding site for PAM, providing the first well defined example of an interaction between an internal Pg-binding region in a protein and a single kringle domain. PMID- 9733733 TI - Contacts between reverse transcriptase and the primer strand govern the transition from initiation to elongation of HIV-1 reverse transcription. AB - HIV-1 reverse transcriptase (RT) utilizes RNA oligomers to prime DNA synthesis. The initiation of reverse transcription requires specific interactions between HIV-1 RNA, primer tRNA3Lys, and RT. We have previously shown that extension of an oligodeoxyribonucleotide, a situation that mimicks elongation, is unspecific and differs from initiation by the polymerization rate and dissociation rate of RT from the primer-template complex. Here, we used replication intermediates to analyze the transition from the initiation to the elongation phases. We found that the 2'-hydroxyl group at the 3' end of tRNA had limited effects on the polymerization and dissociation rate constants. Instead, the polymerization rate increased 3400-fold between addition of the sixth and seventh nucleotide to tRNA3Lys. The same increase in the polymerization rate was observed when an oligoribonucleotide, but not an oligodeoxyribonucleotide, was used as a primer. In parallel, the dissociation rate of RT from the primer-template complex decreased 30-fold between addition of the 17th and 19th nucleotide to tRNA3Lys. The polymerization and dissociation rates are most likely governed by interactions of the primer strand with helix alphaH in the p66 thumb subdomain and the RNase H domain of RT, respectively. PMID- 9733735 TI - Reconstitution of the N-terminal transcription activation function of human mineralocorticoid receptor in a defective human glucocorticoid receptor. AB - N-terminal sequences involved in transcription activation by the human mineralocorticoid receptor (hMR) have yet to be defined. We have addressed this issue and generated overlapping internal deletion mutants hMRDelta59-162, hMRDelta59-247, hMRDelta59-328, hMRDelta162-247, hMRDelta247-328, hMRDelta247 382, and hMRDelta328-382 with intact DNA-binding and hormone-binding domains. A second set of mutant receptors with unique BglII sites was generated to facilitate the isolations of fragments. Immunodetection with anti-hMR peptide antibodies and hormone-binding assays showed that the mutations did not affect the expression of the receptors or ability to bind aldosterone. Distribution of aldosterone binding activity of wild type and deletion mutants expressed in HeLa cells was predominantly nuclear. Furthermore, deletion of sequences between 59 and 390 did not affect DNA binding activity. Transfection studies with HeLa cells revealed a region around residue 247 that was crucial for normal receptor function. Deletion of amino acids 59-162 did not affect the transcriptional activity of the hMR. However, deletion of sequences 247-382 and 328-382 markedly decreased the transcription activation function. The induction of the reporter gene by the chimera hGRDelta71-262/hMR328-382 was 2-fold higher than with the wild type hGR, but 200-fold when compared with hGRDelta71-262, indicating that the AF-1 domain is located between positions 328 and 382 in the hMR. PMID- 9733734 TI - UDP-glucose deficiency causes hypersensitivity to the cytotoxic effect of Clostridium perfringens phospholipase C. AB - A Chinese hamster cell line with a mutation in the UDP-glucose pyrophosphorylase (UDPG:PP) gene leading to UDP-glucose deficiency as well as a revertant cell were previously isolated. We now show that the mutant cell is 10(5) times more sensitive to the cytotoxic effect of Clostridium perfringens phospholipase C (PLC) than the revertant cell. To clarify whether there is a connection between the UDP-glucose deficiency and the hypersensitivity to C. perfringens PLC, stable transfectant cells were prepared using a wild type UDPG:PP cDNA. Clones of the mutant transfected with a construct having the insert in the sense orientation had increased their UDP-glucose level, whereas those of the revertant transfected with a UDPG:PP antisense had reduced their level of UDP-glucose compared with control clones transfected with the vector. Exposure of these two types of transfectant clones to C. perfringens PLC demonstrated that a cellular UDP glucose deficiency causes hypersensitivity to the cytotoxic effect of this phospholipase. Further experiments with genetically engineered C. perfringens PLC variants showed that the sphingomyelinase activity and the C-domain are required for its cytotoxic effect in UDP-glucose-deficient cells. PMID- 9733736 TI - Conserved sequence and structural motifs contribute to the DNA binding and cleavage activities of a geminivirus replication protein. AB - Tomato golden mosaic virus (TGMV), a member of the geminivirus family, has a single-stranded DNA genome that replicates through a rolling circle mechanism in nuclei of infected plant cells. TGMV encodes one essential replication protein, AL1, and recruits the rest of the DNA replication apparatus from its host. AL1 is a multifunctional protein that binds double-stranded DNA, catalyzes cleavage and ligation of single-stranded DNA, and forms oligomers. Earlier experiments showed that the region of TGMV AL1 necessary for DNA binding maps to the N-terminal 181 amino acids of the protein and overlaps the DNA cleavage (amino acids 1-120) and oligomerization (amino acids 134-181) domains. In this study, we generated a series of site-directed mutations in conserved sequence and structural motifs in the overlapping DNA binding and cleavage domains and analyzed their impact on AL1 function in vivo and in vitro. Only two of the fifteen mutant proteins were capable of supporting viral DNA synthesis in tobacco protoplasts. In vitro experiments demonstrated that a pair of predicted alpha-helices with highly conserved charged residues are essential for DNA binding and cleavage. Three sequence motifs conserved among geminivirus AL1 proteins and initiator proteins from other rolling circle systems are also required for both activities. We used truncated AL1 proteins fused to a heterologous dimerization domain to show that the DNA binding domain is located between amino acids 1 and 130 and that binding is dependent on protein dimerization. In contrast, AL1 monomers were sufficient for DNA cleavage and ligation. Together, these results established that the conserved motifs in the AL1 N terminus contribute to DNA binding and cleavage with both activities displaying nearly identical amino acid requirements. However, DNA binding was readily distinguished from cleavage and ligation by its dependence on AL1/AL1 interactions. PMID- 9733737 TI - Defective pancreatic beta-cell glycolytic signaling in hepatocyte nuclear factor 1alpha-deficient mice. AB - Mutations in the hepatocyte nuclear factor-1alpha (HNF-1alpha) gene cause maturity onset diabetes of the young type 3, a form of type 2 diabetes mellitus. In mice lacking the HNF-1alpha gene, insulin secretion and intracellular calcium ([Ca2+]i) responses were impaired following stimulation with nutrient secretagogues such as glucose and glyceraldehyde but normal with non-nutrient stimuli such as potassium chloride. Patch clamp recordings revealed ATP-sensitive K+ currents (KATP) in beta-cells that were insensitive to suppression by glucose but normally sensitive to ATP. Exposure to mitochondrial substrates suppressed KATP, elevated [Ca2+]i, and corrected the insulin secretion defect. NAD(P)H responses to glucose were substantially reduced, and inhibitors of glycolytic NADH generation reproduced the mutant phenotype in normal islets. Flux of glucose through glycolysis in islets from mutant mice was reduced, as a result of which ATP generation in response to glucose was impaired. We conclude that hepatocyte nuclear factor-1alpha diabetes results from defective beta-cell glycolytic signaling, which is potentially correctable using substrates that bypass the defect. PMID- 9733738 TI - Redesign of choline acetyltransferase specificity by protein engineering. AB - Since the development of site-directed mutagenesis techniques over 15 years ago (Zoller, M. J., and Smith, M. (1982) Nucleic Acids Res. 10, 6487-6500), it has been a goal of protein engineering to utilize the procedure to redesign existing enzyme structures to produce proteins with altered or novel catalytic properties. To date, however, the more successful achievements have relied exclusively on the availability of three-dimensional protein structure maps to direct the redesign strategies. Presently, such maps are unavailable for choline acetyltransferase and carnitine acetyltransferase, enzymes that catalyze the reversible transfer of an acetyl group from acetyl-CoA to choline and L-carnitine, respectively. A more empirical approach, based on cross-referencing substrate structure comparisons with protein alignment data, was used to redesign choline acetyltransferase to accommodate L-carnitine as an acceptor of the acetyl group. A mutant choline acetyltransferase that incorporates four amino acid substitutions from wild type, shows a substantial increase in catalytic efficiency (kcat/Km) toward L-carnitine (1,620-fold) and shifts the catalytic discrimination between choline and L carnitine by >390,000 in favor of the latter substrate. These dramatic alterations in catalytic function demonstrate that significant success in protein redesign can be achieved in the absence of three-dimensional protein structure data. PMID- 9733739 TI - Chromatin condensation is not associated with apoptosis. AB - Apoptosis plays an important role in the survival of an organism, and substantial work has been done to understand the signaling pathways that regulate this process. Characteristic changes in chromatin organization accompany apoptosis and are routinely used as markers for cell death. We have examined the organization of chromatin in apoptotic PC12 and HeLa cells by indirect immunofluorescence and electron spectroscopic imaging. Our results indicate that de novo chromatin condensation normally seen during mitosis does not occur when cells undergo apoptosis. Instead, the condensed chromatin typically observed results from aggregation of the heterochromatin. We present evidence that, early in apoptosis, there is a rapid degradation of the nuclease-hypersensitive euchromatin that contains hyperacetylated histones. This occurs coincident with the loss of nuclear integrity due to degradation of lamins and reorganization of intranuclear protein matrix. These events lead to collapse of the nucleus and aggregation of heterochromatin to produce the appearance of condensed apoptotic chromatin. This heterochromatin aggregate is then digested by nucleases to produce the oligonucleosomal DNA ladder that is a hallmark of late apoptosis. Unlike mitosis, we have not seen any evidence for the requirement of phosphorylated histones H1 and H3 to maintain the chromatin in the condensed state. PMID- 9733741 TI - Probing domain functions of chimeric PDE6alpha'/PDE5 cGMP-phosphodiesterase. AB - Chimeric cGMP phosphodiesterases (PDEs) have been constructed using components of the cGMP-binding PDE (PDE5) and cone photoreceptor phosphodiesterase (PDE6alpha') in order to study structure and function of the photoreceptor enzyme. A fully functional chimeric PDE6alpha'/PDE5 enzyme containing the PDE6alpha' noncatalytic cGMP-binding sites, and the PDE5 catalytic domain has been efficiently expressed in the baculovirus/High Five cell system. The catalytic properties of this chimera were practically indistinguishable from those of PDE5, whereas the noncatalytic cGMP binding was similar to that of native purified PDE6alpha'. The inhibitory gamma subunit of PDE6 (Pgamma) enhanced the affinity of cGMP binding at noncatalytic sites of native PDE6alpha' by approximately 6-fold. The polycationic region of Pgamma, Pgamma-24-45, was mainly responsible for this effect, while the inhibitory domain of Pgamma, Pgamma-63-87, was ineffective. On the contrary, Pgamma failed to inhibit catalytic activity of the chimeric PDE6alpha'/PDE5 or to modulate its noncatalytic cGMP binding. Substitutions of Ala residues for the conserved Asn, Asn193 or Asn402, in the two N(K/R)XD-like motifs of the chimeric PDE noncatalytic cGMP-binding sites, each led to a loss of the noncatalytic cGMP binding. Our data suggest that both putative noncatalytic sites of PDE6alpha' are important for binding of cGMP, and that the two binding sites are coupled. Furthermore, mutation Asn402 --> Ala resulted in an approximately 10-fold increase of the Km value for cGMP, indicating that occupation of the noncatalytic cGMP- binding sites of PDE6alpha' may regulate catalytic properties of the enzyme. PMID- 9733740 TI - Evidence that DOCK180 up-regulates signals from the CrkII-p130(Cas) complex. AB - DOCK180 is one of the two principal proteins bound to the SH3 domain of the adaptor protein CrkII. Here, we have studied the involvement of DOCK180 in integrin signaling. DOCK180 was neither phosphorylated nor bound to CrkII in quiescent NIH 3T3 cells and 3Y1 cells. We found that DOCK180 was phosphorylated and bound to CrkII in NIH 3T3 cells stimulated with integrin and also in 3Y1 cells transformed by v-src or v-crk. The binding of DOCK180 to CrkII correlated with the binding of CrkII to p130(Cas), which is a major CrkII SH2 domain-binding protein at focal adhesions. In a reconstitution experiment, expression of DOCK180 induced hyperphosphorylation of p130(Cas) and a concomitant increase in the amount of CrkII bound to p130(Cas). Similarly, binding of DOCK180 to CrkII was also enhanced by the coexpression of p130(Cas). Finally, we found that coexpression of p130(Cas) and CrkII with DOCK180 induced local membrane spreading and accumulation of DOCK180-CrkII-p130(Cas) complexes at focal adhesions. These findings suggest that DOCK180 positively regulates signaling from integrins to CrkII-p130(Cas) complexes at focal adhesions. PMID- 9733742 TI - Nitrosation of uric acid by peroxynitrite. Formation of a vasoactive nitric oxide donor. AB - Peroxynitrite (ONOO-), formed by the reaction between nitric oxide (. NO) and superoxide, has been implicated in the etiology of numerous disease processes. Low molecular weight antioxidants, including uric acid, may minimize ONOO-- mediated damage to tissues. The tissue-sparing effects of uric acid are typically attributed to oxidant scavenging; however, little attention has been paid to the biology of the reaction products. In this study, a previously unidentified uric acid derivative was detected in ONOO--treated human plasma. The product of the uric acid/ONOO- reaction resulted in endothelium-independent vasorelaxation of rat thoracic aorta, with an EC50 value in the range of 0.03-0.3 microM. Oxyhemoglobin, a .NO scavenger, completely attenuated detectable .NO release and vascular relaxation. Uric acid plus decomposed ONOO- neither released .NO nor altered vascular reactivity. Electrochemical quantification of .NO confirmed that the uric acid/ONOO- reaction resulted in spontaneous (thiol-independent) and protracted (t1/2 approximately 125 min) release of .NO. Mass spectroscopic analysis indicated that the product was a nitrated uric acid derivative. The uric acid nitration/nitrosation product may play a pivotal role in human pathophysiology by releasing .NO, which could decrease vascular tone, increase tissue blood flow, and thereby constitute a role for uric acid not previously described. PMID- 9733743 TI - Vanadium K-edge x-ray absorption spectroscopy reveals species differences within the same ascidian genera. A comparison of whole blood from Ascidia nigra and Ascidia ceratodes. AB - Vanadium K-edge x-ray absorption spectroscopy (XAS) was used to examine whole blood preparations from the tunicates Ascidia nigra and Ascidia ceratodes. Each XAS spectrum exhibits a rising edge inflection near 5480 eV characteristic of vanadium(III) and an intensity maximum at 5484.0 eV. In A. ceratodes blood cells, intrinsic aquo-VSO4+ complex ion is indicated by an inflection feature at 5476 eV in the first derivative of the vanadium K-edge XAS spectrum, but this feature is notably absent from the first derivative of the vanadium K-edge spectrum of blood cells from A. nigra. A strong pre-edge feature at 5468.6 eV also uniquely distinguishes the vanadium K-edge XAS spectrum of A. nigra blood cells, implying that vanadyl ion represents approximately 25% of the endogenous vanadium. However, the energy position of the rising edge inflection of the vanadium K-edge XAS spectrum of A. nigra (5479.5 eV) is 1 eV lower than that of A. ceratodes (5480.5 eV), the reverse of any expected shift arising from the endogenous vanadyl ion. Thus, in contrast to A. ceratodes, a significant fraction of the blood cell vanadium(III) in A. nigra is apparently in a ligation environment substantially different from that provided by water. These novel species-related differences may have taxonomic significance. PMID- 9733744 TI - RFX1, a single DNA-binding protein with a split dimerization domain, generates alternative complexes. AB - The transcription of various viral and cellular genes is regulated by palindromic and nonpalindromic DNA sites resembling the EP element of the hepatitis B virus enhancer, which generate similar DNA-protein complexes. The upper EP complex contains homodimers of the transcription regulator RFX1. We show that RFX1 possesses a split, extended dimerization domain composed of several evolutionarily conserved boxes, one of which was previously shown to mediate dimerization. Such an unusually long and complex dimerization domain could potentially serve for generating multiple complexes. In addition to the previously characterized complex, RFX1 generated a novel DNA-protein complex of extremely low mobility, formed only with palindromic DNA sites. Different deletions within the dimerization domain altered the relative abundance of the two complexes, suggesting an interplay between them. Formation of the low mobility complex correlated with transcriptional repression, in that both activities were mediated by several portions of the conserved region. Our results propose a mechanism by which the extended dimerization domain mediates the formation of alternative homodimeric complexes, which differ in the nature of the intersubunit interaction. By participating in different types of interactions, this domain may regulate the relative abundance of the different complexes, thus affecting transcriptional activity. PMID- 9733745 TI - Chimeric receptors composed of phosphoinositide 3-kinase domains and FCgamma receptor ligand-binding domains mediate phagocytosis in COS fibroblasts. AB - Receptors for the Fc portion of IgG (FcgammaR) initiate phagocytosis of IgG opsonized particles by a process involving the assembly of a multi-molecular signaling complex. Several members of this complex have been identified, including Src family kinases, Syk/ZAP 70 family kinases, and phosphoinositide 3 kinase (PI3-K). To test directly the role of PI3-K in mediating phagocytosis, we assessed the phagocytic ability of chimeric receptors composed of FcgammaR extracellular and transmembrane domains fused to regions of the p85 subunit of PI3-K. We found that chimeric receptors with cytoplasmic tails composed of the entire p85 subunit of PI3-K or the inter-Src homology 2 portion of p85 triggered phagocytosis in transfected COS fibroblasts. These two chimeras also showed phosphoinositide kinase activity in vitro when immunoadsorbed. In contrast, a chimera containing only the carboxyl-terminal Src homology 2 domain of p85 that does not interact with the catalytic p110 subunit of PI3-K did not trigger phagocytosis, nor did it show kinase activity in vitro. These data suggest that localization and direct activation of PI3-K at the site of particle attachment is sufficient to trigger the process of phagocytosis. PMID- 9733746 TI - The ortho effect makes manganese(III) meso-tetrakis(N-methylpyridinium-2 yl)porphyrin a powerful and potentially useful superoxide dismutase mimic. AB - The ortho, meta, and para isomers of manganese(III) 5,10,15, 20-tetrakis(N methylpyridyl)porphyrin, MnTM-2-PyP5+, MnTM-3-PyP5+, and MnTM-4-PyP5+, respectively, were analyzed in terms of their superoxide dismutase (SOD) activity in vitro and in vivo. The impact of their interaction with DNA and RNA on the SOD activity in vivo and in vitro has also been analyzed. Differences in their behavior are due to the combined steric and electrostatic factors. In vitro catalytic activities are closely related to their redox potentials. The half-wave potentials (E1/2) are +0.220 mV, +0.052 mV, and +0.060 V versus normal hydrogen electrode, whereas the rates of dismutation (kcat) are 6.0 x 10(7), 4.1 x 10(6), and 3.8 x 10(6) M-1 s-1 for the ortho, meta, and para isomers, respectively. However, the in vitro activity is not a sufficient predictor of in vivo efficacy. The ortho and meta isomers, although of significantly different in vitro SOD activities, have fairly close in vivo SOD efficacy due to their similarly weak interactions with DNA. In contrast, due to a higher degree of interaction with DNA, the para isomer inhibited growth of SOD-deficient Escherichia coli. PMID- 9733747 TI - The Saccharomyces cerevisiae NDE1 and NDE2 genes encode separate mitochondrial NADH dehydrogenases catalyzing the oxidation of cytosolic NADH. AB - In Saccharomyces cerevisiae, the NDI1 gene encodes a mitochondrial NADH dehydrogenase, the catalytic side of which projects to the matrix side of the inner mitochondrial membrane. In addition to this NADH dehydrogenase, S. cerevisiae exhibits another mitochondrial NADH-dehydrogenase activity, which oxidizes NADH at the cytosolic side of the inner membrane. To investigate whether open reading frames YMR145c/NDE1 and YDL 085w/NDE2, which exhibit sequence similarity with NDI1, encode the latter enzyme, NADH-dependent mitochondrial respiration was assayed in wild-type S. cerevisiae and nde deletion mutants. Mitochondria were isolated from aerobic, glucose-limited chemostat cultures grown at a dilution rate (D) of 0. 10 h-1, in which reoxidation of cytosolic NADH by wild-type cells occurred exclusively by respiration. Compared with the wild type, rates of mitochondrial NADH oxidation were about 3-fold reduced in an nde1Delta mutant and unaffected in an nde2Delta mutant. NADH-dependent mitochondrial respiration was completely abolished in an nde1Delta nde2Delta double mutant. Mitochondrial respiration of substrates other than NADH was not affected in nde mutants. In shake flasks, an nde1Delta nde2Delta mutant exhibited reduced specific growth rates on ethanol and galactose but not on glucose. Glucose metabolism in aerobic, glucose-limited chemostat cultures (D = 0.10 h-1) of an nde1Delta nde2Delta mutant was essentially respiratory. Apparently, under these conditions alternative systems for reoxidation of cytosolic NADH could replace the role of Nde1p and Nde2p in S. cerevisiae. PMID- 9733748 TI - Prodomain-dependent nuclear localization of the caspase-2 (Nedd2) precursor. A novel function for a caspase prodomain. AB - Caspases are cysteine proteases that play an essential role in apoptosis by cleaving several key cellular proteins. Despite their function in apoptosis, little is known about where in the cell they are localized and whether they are translocated to specific cellular compartments upon activation. In the present paper, using Aequorea victoria green fluorescent protein fusion constructs, we have determined the localization of Nedd2 (mouse caspase-2) and show that both precursor and processed caspase-2 localize to the cytoplasmic and the nuclear compartments. We demonstrate that the nuclear localization of caspase-2 is strictly dependent on the presence of the prodomain. A caspase-2 prodomain-green fluorescent protein localized to dot- and fiber-like structures mostly in the nucleus, whereas a protein lacking the prodomain was largely concentrated in the cytoplasm. We also show that an amino-terminal fusion of the prodomain of caspase 2 to caspase-3 mediates nuclear transport of caspase-3, which is normally localized in the cytoplasm. These results suggest that, in addition to roles in dimerization and recruitment through adaptors, the caspase-2 prodomain has a novel function in nuclear transport. PMID- 9733749 TI - Human minichromosome maintenance proteins and human origin recognition complex 2 protein on chromatin. AB - Minichromosome maintenance (Mcm) proteins and the constituents of the origin recognition complex (Orc) are essential components of the eukaryotic replication initiation apparatus. Published evidence strongly suggests that the binding of Mcm proteins to chromatin is contingent upon the prior binding of Orc proteins. Here we use two different approaches to investigate the presence of the human ORC2 protein and of Mcm proteins on chromatin of HeLa cells in various cell cycle phases. First, we mobilized chromatin-bound proteins by micrococcal nuclease and analyzed the resulting digestion products by sucrose gradient centrifugations. Under digestion conditions when Mcm proteins were almost entirely released from chromatin, ORC2 protein was found to be associated with chromatin fragments containing several hundred base pairs of DNA. Second, we used an in vivo cross linking procedure to covalently link Mcm proteins and ORC2 to DNA by short exposure of intact HeLa cells to formaldehyde. Specific immunoprecipitations revealed that cross-linked nucleoprotein fragments carried either Mcm proteins or ORC2 protein, but not both. Based on the lengths of the DNA fragments in immunoprecipitates, we estimate that the distance between chromatin-bound ORC2 protein and chromatin-bound Mcm proteins must be at least 500-1000 base pairs in HeLa cells. PMID- 9733750 TI - ATP binding to the Escherichia coli clamp loader powers opening of the ring shaped clamp of DNA polymerase III holoenzyme. AB - The Escherichia coli gamma complex serves as a clamp loader, catalyzing ATP dependent assembly of beta protein clamps onto primed DNA templates during DNA replication. These ring-shaped clamps tether DNA polymerase III holoenzyme to the template, facilitating rapid and processive DNA synthesis. This report focuses on the role of ATP binding and hydrolysis catalyzed by the gamma complex during clamp loading. We show that the energy from ATP binding to gamma complex powers several initial events in the clamp loading pathway. The gamma complex (gamma2 delta delta'chi psi) binds two ATP molecules (one per gamma subunit in the complex) with high affinity (Kd = 1-2. 5 x 10(-6) M) or two adenosine 5'-O-(3 thiotriphosphate)(ATPgammaS) molecules with slightly lower affinity (Kd = 5-6.5 x 10(-6) M). Experiments performed prior to the first ATP turnover (kcat = 4 x 10( 3) s-1 at 4 degreesC), or in the presence of ATPgammaS (kcat = 1 x 10(-4) s-1 at 37 degreesC), demonstrate that upon interaction with ATP the gamma complex undergoes a change in conformation. This ATP-bound gamma complex binds beta and opens the ring at the dimer interface. Still prior to ATP hydrolysis, the composite of gamma complex and the open beta ring binds with high affinity to primer-template DNA. Thus ATP binding powers all the steps in the clamp loading pathway leading up to the assembly of a gamma complex. open beta ring.DNA intermediate, setting the stage for ring closing and turnover of the clamp loader, steps that may be linked to subsequent hydrolysis of ATP. PMID- 9733751 TI - Pre-steady state analysis of the assembly of wild type and mutant circular clamps of Escherichia coli DNA polymerase III onto DNA. AB - The beta protein, a dimeric ring-shaped clamp essential for processive DNA replication by Escherichia coli DNA polymerase III holoenzyme, is assembled onto DNA by the gamma complex. This study examines the clamp loading pathway in real time, using pre-steady state fluorescent depolarization measurements to investigate the loading reaction and ATP requirements for the assembly of beta onto DNA. Two beta dimer interface mutants, L273A and L108A, and a nonhydrolyzable ATP analog, adenosine 5'-O-(3-thiotriphosphate) (ATPgammaS), have been used to show that ATP binding is required for gamma complex and beta to associate with DNA, but that a gamma complex-catalyzed ATP hydrolysis is required for gamma complex to release the beta.DNA complex and complete the reaction. In the presence of ATP and gamma complex, the beta mutants associate with DNA as efficiently as wild type beta. However, completion of the reaction is much slower with the beta mutants because of decreased ATP hydrolysis by the gamma complex, resulting in a much slower release of the mutants onto DNA. The effects of mutations in the dimer interface were similar to the effects of replacing ATP with ATPgammaS in reactions using wild type beta. Thus, the assembly of beta around DNA is coupled tightly to the ATPase activity of the gamma complex, and completion of the assembly process requires ATP hydrolysis for turnover of the catalytic clamp loader. PMID- 9733752 TI - Identification and characterization of a sialidase released by the salivary gland of the hematophagous insect Triatoma infestans. AB - Sialidases (EC 3.2.1.18) are commonly found in viruses, bacteria, fungi, protozoa, and vertebrates, but not in invertebrates. We have previously reported the presence of a new sialidase activity in the gut of exclusively hematophagous insects of the Triatoma genus, which transmit Chagas' disease (Amino, R., Acosta, A., Morita, O. M., Chioccola, V. L. P., and Schenkman, S. (1995) Glycobiology 5, 625-631). Here we show that this sialidase is present in the salivary gland of Triatoma infestans, and it is released with the saliva during the insect bite. The sialidase was purified to homogeneity (>5000 times) to a specific activity of more than 20 units/mg. It elutes from a gel filtration column with a volume corresponding to the size of 33 kDa, and it migrates as a single 26-kDa band in SDS-polyacrylamide gel electrophoresis, which is unusually smaller when compared with other known sialidases. T. infestans sialidase hydrolyzes preferentially alpha2-->3-linked sialic acids at pH 4-8, with maximal activity between pH 5.5 and 6.5, which is compatible with the optimal pH of secreted sialidases. The sialidase is competitively inhibited by 2-deoxy-2, 3-dehydro-N-acetyl-neuraminic acid (Ki = 0.075 mM) and differently from many sialidases, with exception of Salmonella typhimurium sialidase, it is inhibited competitively by HEPES (Ki = 15 mM). The fact that T. infestans sialidase is released with the saliva and can hydrolyze sialyl-LewisX blood groups, which are the ligands for selectins, suggests that it might have a role in the blood feeding. PMID- 9733753 TI - Mechanisms of spectral tuning in blue cone visual pigments. Visible and raman spectroscopy of blue-shifted rhodopsin mutants. AB - Spectral tuning by visual pigments involves the modulation of the physical properties of the chromophore (11-cis-retinal) by amino acid side chains that compose the chromophore-binding pocket. We identified 12 amino acid residues in the human blue cone pigment that might induce the required green-to-blue opsin shift. The simultaneous substitution of nine of these sites in rhodopsin (M86L, G90S, A117G, E122L, A124T, W265Y, A292S, A295S, and A299C) shifted the absorption maximum from 500 to 438 nm, accounting for 2,830 cm-1, or 80%, of the opsin shift between rhodopsin and the blue cone pigment. Raman spectroscopy of mutant pigments shows that the dielectric character and architecture of the chromophore binding pocket are specifically altered. An increase in the number of dipolar side chains near the protonated Schiff base of retinal increases the ground excited state energy gap via long range dipole-dipole Coulomb interaction. In addition, the W265Y substitution causes a decrease in solvent polarizability near the chromophore ring structure. Finally, two substitutions on transmembrane helix 3 (A117G and E122L) act in combination with the other substitutions to alter the binding-pocket structure, resulting in stronger interaction of the protonated Schiff base group with the surrounding dipolar groups and the counterion. Taken together, these results identify the amino acid side chains and the underlying physical mechanisms responsible for a majority of the opsin shift in blue visual pigments. PMID- 9733754 TI - Regulated endocytosis of G-protein-coupled receptors by a biochemically and functionally distinct subpopulation of clathrin-coated pits. AB - beta-2 Adrenergic receptors (B2ARs) are endocytosed by clathrin-coated pits. This process serves specialized functions in signal transduction and receptor regulation, raising the question of whether B2ARs are associated with biochemically specialized membrane vesicles during their endocytic trafficking. Here we show that B2ARs are endocytosed by a distinct subpopulation of clathrin coated pits, which represent a limited subset of coated pits present in the plasma membrane, even in cells overexpressing both B2ARs and beta-arrestin. Coated pits mediating agonist-induced endocytosis of B2ARs differ from other coated pits mediating constitutive endocytosis of transferrin receptors in their temperature dependence for fission from the plasma membrane and in the association of their membrane coats with beta-arrestin. Endocytosis of these coated pits generates endocytic vesicles selectively enriched in B2ARs, which fuse within approximately 10 min after their formation with a common population of endosomes containing both B2ARs and transferrin receptors. These observations demonstrate, for the first time, the existence of a functionally and biochemically distinct subpopulation of clathrin-coated pits that mediate the agonist-regulated endocytosis of G-protein-coupled receptors, and they suggest a new model for the formation of compositionally specialized membrane vesicles at the earliest stage of the endocytic pathway. PMID- 9733755 TI - Detailed comparison of two molecular models of the human CD40 ligand with an x ray structure and critical assessment of model-based mutagenesis and residue mapping studies. AB - The interactions between the B cell receptor CD40 and its ligand on T cells are critical for the integrity of immune responses. The human CD40 ligand gp39, a tumor necrosis factor-like protein, has been the subject of intense efforts to identify the receptor-binding site and to analyze naturally occurring mutations that compromise gp39 function in vivo. These investigations relied heavily on molecular models of gp39, built in the presence of only approximately 25% sequence identity to tumor necrosis factor. The x-ray structure of gp39 has made it possible to assess modeling accuracy and to evaluate the results of model based mutagenesis analyses. Although the models display local errors, their accuracy was sufficient to predict the CD40-binding site, to map natural mutations, and to rationalize their effects. One of five gp39 residues critical for CD40 binding was displaced in the models, and 1 of 21 point mutants was incorrectly classified. Factors most important for the reliability of the molecular models and their successful applications were valid sequence alignments and the focus of experimental studies on regions of high prediction confidence. Analysis of mutagenesis experiments correlated with anti-gp39 monoclonal antibody binding studies to assess the conformational integrity of mutant proteins. PMID- 9733756 TI - Differential activation of p70 and p85 S6 kinase isoforms during cardiac hypertrophy in the adult mammal. AB - An adult feline right ventricular pressure overload (RVPO) model was used to examine the two S6 kinase (S6K) isoforms, p70(S6K) and p85(S6K), that are involved in translational and transcriptional activation. Biochemical and confocal microscopy analyses at the level of the cardiocyte revealed that p70(S6K) is present predominantly in the cytosol, substantially activated in 1-h RVPO (>12 fold), and phosphorylated in the pseudosubstrate domain at the Ser-411, Thr-421, and Ser-424 sites. p85(S6K), which was localized exclusively in the nucleus, showed activation subsequent to p70(S6K), with a sustained increase in phosphorylation for up to 48 h of RVPO at equivalent sites of p70(S6K), Thr-421 and Ser-424, but not at Ser-411. Neither isoform translocated between the cytosol and the nucleus. Further studies to determine potential upstream elements of S6K activation revealed: (i) similar time course of activation for protein kinase C isoforms (alpha, gamma, and epsilon) and c-Raf, (ii) absence of accompanying phosphatidylinositol 3-kinase activation, (iii) activation of c-Src subsequent to p70(S6K), and (iv) similar changes in adult cardiocytes after treatment with 12-O tetradecanoylphorbol-13-acetate. Thus, these studies suggest that a protein kinase C-mediated pathway couples pressure overload to growth induction via differential activation of S6K isoforms in cardiac hypertrophy. PMID- 9733757 TI - Expression of carbonic anhydrase V in pancreatic beta cells suggests role for mitochondrial carbonic anhydrase in insulin secretion. AB - Carbonic anhydrase V (CA-V) is a mitochondrial enzyme that provides bicarbonate for pyruvate carboxylase in liver and kidney. In the course of a survey of the tissue distribution of CA-V, we detected intense immunostaining in pancreatic islets when sections from rat and mouse pancreases were reacted with a polyclonal antibody to recombinant mouse CA-V. The distribution and large number of CA-V positive cells in each islet suggested that they represented beta cells. Double immunofluorescence staining of tissue sections and isolated islet cells showed cellular colocalization of CA-V and insulin, confirming that beta cells contain CA-V. Western blotting of rat islets of Langerhans and primary beta cells showed 33- and 30-kDa polypeptides of precursor and mature CA-V, respectively. The CA-V expression was beta cell-specific since no CA-V immunoreaction was detected in the primary alpha cells. Immunohistochemical staining for CA-I, CA-II, CA-IV, CA VI, and CA-IX was negative in beta cells, and Western blotting of beta cells also failed to identify any CA in beta cells except CA-V. The specific localization of CA-V in beta cells led us to hypothesize that CA-V may be functionally linked to the regulation of insulin secretion. Consistent with this hypothesis, the CA inhibitor acetazolamide was found to be a strong inhibitor of glucose-stimulated insulin secretion by isolated rat pancreatic islets. PMID- 9733758 TI - Differential coupling of alpha1-, alpha2-, and beta-adrenergic receptors to mitogen-activated protein kinase pathways and differentiation in transfected PC12 cells. AB - Three adrenergic receptor families that selectively activate three different G proteins (alpha1/Gq/11, alpha2/Gi, and beta/Gs) were used to study mitogen activated protein kinase (MAPK) activation and differentiation in PC12 cells. PC12 cells were stably transfected with alpha1A-, alpha2A-, or beta1-adrenergic receptors (ARs) in an inducible expression vector, and subclones were characterized. Norepinephrine stimulated inositol phosphate formation in alpha1A transfected cells, inhibited cyclic adenosine 3'5'-monophosphate (cAMP) formation in alpha2A-transfected cells, and stimulated cAMP formation in beta1-transfected cells. Nerve growth factor activated extracellular signal-regulated kinases (ERKs) in all cell lines; however, norepinephrine activated ERKs only in alpha1A- and beta1-transfected cells but not in alpha2A-transfected cells. Norepinephrine also activated c-Jun NH2-terminal kinase and p38 MAPK in alpha1A-transfected cells but not in beta1- or alpha2A-transfected cells. Norepinephrine caused differentiation of PC12 cells expressing alpha1A-ARs but not those expressing beta1- or alpha2A-ARs. However, norepinephrine acted synergistically with nerve growth factor in promoting differentiation of cells expressing beta1-ARs. Whereas ERKs are activated by Gi- but not Gs-linked receptors in many fibroblastic cell lines, we observed the opposite in PC12 cells. The results show that activation of the different G protein signaling pathways has different effects on MAPKs and differentiation in PC12 cells, with Gq signaling pathways activating all three major MAPK pathways. PMID- 9733759 TI - The herpes simplex virus type 1 origin binding protein. Specific recognition of phosphates and methyl groups defines the interacting surface for a monomeric DNA binding domain in the major groove of DNA. AB - The UL9 gene of herpes simplex virus type 1 (HSV-1) encodes an origin binding protein (OBP). It is an ATP-dependent DNA helicase and a sequence-specific DNA binding protein. The latter function is carried out by the C-terminal domain of OBP (DeltaOBP). We have now performed a quantitative analysis of the interaction between DeltaOBP and its recognition sequence, GTTCGCAC, in oriS. Initially optimal conditions for binding were carefully determined. We observed that complexes with different electrophoretic mobilities were formed. A cross-linking experiment demonstrated that nonspecific complexes containing 2 or more protein monomers per DNA molecule were formed at high protein concentrations. The specific complex formed at low concentrations of DeltaOBP had an electrophoretic mobility corresponding to a 1:1 complex. We then demonstrated that the methyl groups of thymine in the major groove were essential for high affinity binding. Changes in the minor groove had considerably smaller effects. Ethylation interference experiments indicated that specific contacts were made between OBP and three phosphates in the recognition sequence. Finally, these observations were used to present a model of the surface of DNA that interacts with DeltaOBP in a sequence-specific manner. PMID- 9733760 TI - The GAGA factor of Drosophila binds triple-stranded DNA. AB - The Drosophila GAGA factor binds specifically to simple repeating d(GA.TC)n DNA sequences. These sequences are known to be capable of forming triple-stranded DNA as well as other non-B-DNA conformations. Here, it is shown that GAGA binds to a d[CT(GA.TC)]22 intermolecular triplex with similar specificity and affinity as to a regular double-stranded B-form d(GA.TC)22 sequence. The interaction of GAGA with triplex DNA cannot, however, stimulate transcription in vitro. The affinity of GAGA for triplexes of the purine motif, such as a d[AG(GA.TC)]22 intermolecular triplex, is significantly lower. The DNA binding domain of GAGA is sufficient for efficient binding to triplex DNA. Based on the reported solution structure of the complex of GAGA-DNA binding domain with double-stranded DNA, a model for its interaction with triplex DNA is proposed in which most of the protein-DNA contacts observed in duplex DNA are maintained, especially those occurring through the minor groove. The higher negative charge of the triplex is likely to have also an important contribution to both the specificity and affinity of the interaction. PMID- 9733761 TI - Regulated Co-translational ubiquitination of apolipoprotein B100. A new paradigm for proteasomal degradation of a secretory protein. AB - Presentation of a wild-type secretory protein, apolipoprotein B100 (apoB), to the cytosol for ubiquitin-proteasome proteolysis has been observed in HepG2 cells. A currently accepted model for proteasomal degradation of secretory proteins is retrograde translocation of the substrate polypeptides from the lumen of endoplasmic reticulum (ER) back to the cytosol. In this report, we present evidence that newly synthesized apoB becomes exposed to the cytosol and targeted to the proteasomes in a co-translational manner. Thus, after protein translation was synchronized with puromycin, partially synthesized apoB polypeptides were found to be conjugated to ubiquitin. The magnitude of co-translational ubiquitination and subsequent degradation of apoB was increased when cells were pretreated with either herbimycin A to induce cytosolic Hsp70 or with an inhibitor of microsomal triglyceride transfer protein; both treatments impede translocation of nascent apoB across the ER membrane. These treatments also decreased secretion of apoB and increased its degradation via the ubiquitin proteasome pathway. We suggest that translocation arrest with subsequent co translational exposure to the cytosol provides an alternative model to explain how mammalian secretory proteins can overcome topological segregation by the ER membrane and undergo degradation by the ubiquitin-proteasome pathway. PMID- 9733762 TI - Dictyostelium TRFA homologous to yeast Ssn6 is required for normal growth and early development. AB - The TPR (tetratricopeptide repeat) family became widespread during evolution, having been found from bacteria to mammals. By means of restriction enzyme mediated integration, we have identified a Dictyostelium gene (trfA) highly homologous to a Saccharomyces cerevisiae gene encoding a TPR protein, Ssn6 (Cyc8), which functions as a global transcriptional repressor for diverse genes. The deduced amino acid sequence of the Dictyostelium gene product, TRFA, contains 10 consecutive TPR units as well as Gln repeats, Asn repeats, and a region rich in Glu, Lys, Ser, and Thr. The sequences of some of the 10 TPR units in TRFA are more than 70% identical to the corresponding units in Ssn6. The trfA- cells produced smooth plaques on a bacterial lawn and failed to aggregate normally when starved on a plain agar plate. Individual trfA- cells also failed to correctly respond to cAMP, although the adenylyl cyclase of trfA- cells was expressed upon starvation and activated by stimulation with cAMP as in the wild-type cells. When cultured in a rich medium in suspension, they grew more slowly and stopped growing at a lower density than the wild-type cells. Furthermore, they divided into cells of various sizes and tended to be much smaller than the wild-type cells. These pleiotropic defects of the trfA- cells suggest the possibility that Dictyostelium TRFA may regulate the transcription of diverse genes required for normal growth and early development. PMID- 9733763 TI - Crystal structure of polygalacturonase from Erwinia carotovora ssp. carotovora. AB - The crystal structure of the 40-kDa endo-polygalacturonase from Erwinia carotovora ssp. carotovora was solved by multiple isomorphous replacement and refined at 1.9 A to a conventional crystallographic R-factor of 0.198 and Rfree of 0.239. This is the first structure of a polygalacturonase and comprises a 10 turn right-handed parallel beta-helix domain with two loop regions forming a "tunnel like" substrate-binding cleft. Sequence conservation indicates that the active site of polygalacturonase is between these two loop regions, and comparison of the structure of polygalacturonase with that of rhamnogalacturonase A from Aspergillus aculeatus enables two conserved aspartates, presumed to be catalytic residues, to be identified. An adjacent histidine, in accord with biochemical results, is also seen. A similarity in overall electrostatic properties of the substrate-binding clefts of polygalacturonase and pectate lyase, which bind and cleave the same substrate, polygalacturonic acid, is also revealed. PMID- 9733764 TI - Overexpression of perilipin A and B blocks the ability of tumor necrosis factor alpha to increase lipolysis in 3T3-L1 adipocytes. AB - Perilipins, a family of phosphoproteins, are specifically located at the surface of intracellular lipid (triacylglycerol) droplets, the site of lipolysis. Stimulation of lipolysis in 3T3-L1 adipocytes by tumor necrosis factor alpha (TNF alpha) is associated with a decrease in total cellular expression of perilipin A and B, consistent with the hypothesis that a decrease in perilipin protein expression is required for TNF-alpha-induced lipolysis. Adenovirus-mediated overexpression of perilipin A or B maintains perilipin protein levels on the lipid droplet and blocks TNF-alpha-induced lipolysis. In contrast, overexpression of perilipin A or perilipin B does not inhibit isoproterenol-stimulated lipolysis and does not alter the isoproterenol-induced migration of perilipins from the lipid droplet. These results provide the first evidence of how perilipin functions and suggest that TNF-alpha regulates lipolysis, in part, by decreasing perilipin protein levels at the lipid droplet surface. PMID- 9733765 TI - An enhancer element in the EphA2 (Eck) gene sufficient for rhombomere-specific expression is activated by HOXA1 and HOXB1 homeobox proteins. AB - In the hindbrain of the mouse embryo, there is often coincident rhombomere restricted expression of Eph receptor tyrosine kinases and Hox homeobox genes, raising the possibility of regulatory interactions. In this paper, we have identified cis-acting regulatory sequences of the EphA2 (Eck) gene, which direct node and hindbrain-specific expression in transgenic embryos. An 8-kilobase region of mouse genomic DNA element was sufficient to drive rhombomere 4 (r4) specific expression while conferring patchy expression in the node. Further analysis localized the rhombomere-specific enhancer to a 0.9-kilobase sequence. This element contains multiple Hox-Pbx consensus binding sites that bind to both HOXA1/Pbx1 and HOXB1/Pbx1 proteins in vitro. Co-expression of either HOXA1 or HOXB1 with Pbx1 transactivated EphA2 enhancer-dependent reporter gene expression. These results, together with observations of reduced EphA2 expression in hoxa1 and hoxb1 double mutant mice, suggest that expression of EphA2 gene in rhombomere 4 is directly regulated by Hoxa1 and Hoxb1 homeobox transcription factors. PMID- 9733766 TI - The cyclophilin-like domain mediates the association of Ran-binding protein 2 with subunits of the 19 S regulatory complex of the proteasome. AB - The combination of the Ran-binding domain 4 and cyclophilin domains of Ran binding protein 2 selectively associate with a subset of G protein-coupled receptors, red/green opsins, upon cis-trans prolyl isomerase-dependent and direct modification of opsin followed by association of the modified opsin isoform to Ran-binding domain 4. This effect enhances in vivo the production of functional receptor and generates an opsin isoform with no propensity to self-aggregate in vitro. We now show that another domain of Ran-binding protein 2, cyclophilin-like domain, specifically associates with the 112-kDa subunit, P112, and other subunits of the 19 S regulatory complex of the 26 S proteasome in the neuroretina. This association possibly mediates Ran-binding protein 2 limited proteolysis into a smaller and stable isoform. Also, the interaction of Ran binding protein 2 with P112 regulatory subunit of the 26 S proteasome involves still another protein, a putative kinesin-like protein. Our results indicate that Ran-binding protein 2 is a key component of a macro-assembly complex selectively linking protein biogenesis with the proteasome pathway and, thus, with potential implications for the presentation of misfolded and ubiquitin-like modified proteins to this proteolytic machinery. PMID- 9733767 TI - AP-1 and ets transcription factors regulate the expression of the human SPRR1A keratinocyte terminal differentiation marker. AB - The 173-base pair proximal promoter of SPRR1A is necessary and sufficient for regulated expression in primary keratinocytes induced to differentiate either by increasing extracellular calcium or by 12-O-tetradecanoylphorbol-13-acetate (TPA) treatment. Whereas calcium-induced expression depends both on an AP-1 and an Ets binding site in this region, responsiveness to TPA resides mainly (but not exclusively) on the Ets element, indicating that Ets factors are important targets for protein kinase C signaling during keratinocyte terminal differentiation. This conclusion is further substantiated by the finding that expression of ESE-1, an Ets transcription factor involved in SPRR regulation, is also induced by TPA, with kinetics similar to SPRR1A. The strict AP-1 requirement in SPRR1A for calcium-induced differentiation is not found for SPRR2A, despite the presence of an identical AP-1 consensus binding site in this gene. Binding site swapping indicates that both the nucleotides flanking the TGAGTCA core sequence and the global promoter context are essential in determining the contribution of AP-1 factors in gene expression during keratinocyte terminal differentiation. In the distal SPRR1A promoter region, a complex arrangement of positive and negative regulatory elements, which are only conditionally needed for promoter activity, are likely involved in gene-specific fine-tuning of the expression of this member of the SPRR gene family. PMID- 9733769 TI - The insulin-like growth factor (IGF)binding protein 1 binding epitope on IGF-I probed by heteronuclear NMR spectroscopy and mutational analysis. AB - NMR spectroscopy studies and biosensor interaction analysis of native and site directed mutants of insulin-like growth factor I (IGF-I) was applied to identify the involvement of individual residues in IGF-I binding to IGF-binding protein 1 (IGFBP-1). Backbone NMR chemical shifts were found to be affected by IGFBP-1 binding in the following residues: Pro2, Glu3, Cys6, Gly7, Gly19, Pro28-Gly30, Gly32, Arg36, Arg37, Gln40-Gly42, Pro63, Lys65, Pro66, and Lys68-Ala70. Three IGF I arginine side chains were identified by NMR to participate in IGFBP-1 binding. All IGF-I arginine residues were replaced by alanines, using site-directed mutagenesis, in four single substituted variants, IGF-I(R21A), IGF-I(R50A), IGF I(R55A), and IGF-I(R56A), and one double replacement mutant, IGF-I(R36A/R37A). Biosensor interaction analysis binding studies demonstrate the involvement of Arg36-Arg37 and Arg50 in IGFBP-1 binding, while experiments with the IGF-I receptor implicate Arg21, Arg36-Arg37, and Arg56 as part of the receptor binding epitope. These overlapping binding surfaces explain why IGF-I receptor and IGFBP 1 binding to IGF-I is competitive. The C terminus of free, but not IGFBP-1-bound, IGF-I is found to exist in two distinct, NMR-detectable conformations at 30 degreesC. One possible explanation for this structural heterogeneity could be cis trans isomerization of the Cys6-Cys48 disulfide bond. PMID- 9733768 TI - Identification and characterization of novel clathrin adaptor-related proteins. AB - We have identified a human approximately 87-kDa protein, designated as gamma2 adaptin, that is similar to gamma-adaptin (called gamma1-adaptin in this paper), a large chain of the AP-1 clathrin-associated adaptor complex, not only in the primary structure (60% amino acid identity) but also in the domain organization. Northern blot analysis has shown that its mRNA is expressed in a variety of tissues. Analysis using a yeast two-hybrid system has revealed that, similarly to gamma1-adaptin, gamma2-adaptin is capable of interacting not only with the sigma1 chain (called as sigma1A in this paper), the small chain of the AP-1 complex, but also with a novel sigma1-like protein, designated as sigma1B, which shows an 87% amino acid identity to sigma1A; and that, unlike gamma1-adaptin, it is unable to interact with beta1-adaptin, another large chain of the AP-1 complex. Immunofluorescence microscopy analysis has revealed that gamma2-adaptin is localized to paranuclear vesicular structures that are not superimposed on structures containing gamma1-adaptin. Furthermore, unlike gamma1-adaptin, gamma2 adaptin is recruited onto membranes in the presence of a fungal antibiotic, brefeldin A. These data suggest that gamma2-adaptin constitute a novel adaptor related complex that participates in a transport step different from that of AP 1. PMID- 9733770 TI - Requirement for an interaction of XRCC4 with DNA ligase IV for wild-type V(D)J recombination and DNA double-strand break repair in vivo. AB - The XRCC4 gene is required for the repair of DNA double-strand breaks in mammalian cells. Without XRCC4, cells are hypersensitive to ionizing radiation and deficient for V(D)J recombination. It has been demonstrated that XRCC4 binds and stimulates DNA ligase IV, which has led to the hypothesis that DNA ligase IV is essential for both of these processes. In this study deletion mutants of XRCC4 were tested for their ability to associate with DNA ligase IV in vitro and for their ability to reconstitute XRCC4-deficient cells in vivo. We find that a central region of XRCC4 from amino acids 100-250 is necessary for DNA ligase IV binding and that deletions within this region functionally inactivates XRCC4. Deletions within the C-terminal 84 amino acids neither affect DNA ligase IV binding nor the in vivo function of XRCC4. The correlation between the ability or inability of XRCC4 to bind DNA ligase IV and its ability or failure to reconstitute wild-type DNA repair in vivo, respectively, demonstrates for the first time that the physical interaction with DNA ligase IV is crucial for the in vivo function of XRCC4. Deletions within the N-terminal 100 amino acids inactivate XRCC4 in vivo but leave DNA ligase IV binding unaffected. This indicates further DNA ligase IV-independent functions of XRCC4. PMID- 9733771 TI - The crystal structure of amyloidogenic Leu55 --> Pro transthyretin variant reveals a possible pathway for transthyretin polymerization into amyloid fibrils. AB - The x-ray crystal structure of the amyloidogenic Leu55 --> Pro transthyretin (TTR) variant, implicated as the causative agent in early-onset familial amyloidotic polyneuropathy (Jacobson, D. R., McFarlin, D. E., Kane, I., and Buxbaum, J. N. (1992) Hum. Genet. 89, 353-356), has been solved by molecular replacement, refined at 2.7 A to a Rcryst value of 0.190 (Fobs > 2.0sigma), and compared with wild-type transthyretin to understand the molecular mechanism(s) involved in amyloidogenesis. Leu55 --> Pro TTR crystallizes in space group C2, with eight monomers in the asymmetric unit, and the observed packing contacts are considerably different from those described for the wild-type protein. Refinement of the crystal structure shows that the proline for leucine substitution disrupts the hydrogen bonds between strands D and A, resulting in different interface contacts. Based on the assumption that the observed packing contacts may be significant for amyloidogenesis, a model for the TTR amyloid is proposed. It consists of a tubular structure with inner and outer diameters approximately of 30 and 100 A and four monomers per cross-section. PMID- 9733772 TI - Specific contribution of Tyk2 JH regions to the binding and the expression of the interferon alpha/beta receptor component IFNAR1. AB - Cytokine signaling involves the activation of the Janus kinase (JAK) family of tyrosine kinases. These enzymes are physically associated with cytokine receptor components. Here, we sought to define the molecular basis of the interaction between Tyk2 and IFNAR1, a component of the interferon alpha/beta receptor, by delimiting a minimal IFNAR1 binding region in the Tyk2 protein. Using an in vitro assay system, we narrowed down the interaction domain to a region comprising the JH7 and part of the JH6 homology boxes (amino acids 22-221). When expressed in Tyk2-negative cells, the JH7-6 region was unable to stabilize IFNAR1 protein levels, a critical function that we previously attributed to the N region (amino acids 1-591) of Tyk2. Moreover, substitution of the JH7-JH6 domain in JAK1 with that of Tyk2 did not restore IFNAR1 level nor interferon alpha signaling in Tyk2 negative cells. Thus, the major interaction surface lies within JH7-6, but additional JH regions (JH5-4-3) contribute in a specific manner to the in vivo assembly of Tyk2 and IFNAR1. Evidence is also provided of the lack of specificity of the Tyk2 kinase-like and tyrosine kinase domains in interferon alpha/beta receptor signaling. PMID- 9733773 TI - Preferential binding of high mobility group 1 protein to UV-damaged DNA. Role of the COOH-terminal domain. AB - Binding of chromosomal high mobility group 1 protein (HMG1) to UV-damaged DNA has been studied with oligonucleotides containing a single dipyrimidine site for formation of UV photolesions. Irradiation of an oligonucleotide with unique TT dinucleotide resulted in generation of cyclobutane pyrimidine dimer with no evidence for induction of (6-4) photoproducts, whereas the analysis of irradiated TC-containing oligonucleotide detected (6-4) photoproducts but not cyclobutane pyrimidine dimers. Mobility shift assays have revealed that HMG1 protein binds preferentially to irradiated TT and TC oligonucleotides. Photoreversal of cyclobutane pyrimidine dimers with DNA photolyase and hydrolysis of the (6-4) photoproducts with hot alkali substantially reduced but did not eliminate binding of HMG1. The protein, therefore, appears to bind the two main types of UV damages in DNA, but some other photolesion(s) contributes to the preferential binding of HMG1 to irradiated DNA. By quantifying gel shift assays and considering the efficiencies of lesion formation, we determined dissociation constants of 1.2 +/- 0.5 and 4.0 +/- 1.5 microM for irradiated TT and TC oligonucleotides, respectively, and 70 +/- 20 microM for the control non-irradiated probes. Tryptic removal of the acidic COOH-terminal domain of HMG1 significantly affected binding of the protein to both irradiated and intact oligonucleotides. The potential role of HMG1 in recognition of the UV lesions in DNA is discussed. PMID- 9733774 TI - Molecular characterization of a broad selectivity neutral solute channel. AB - In all living cells, coordination of solute and water movement across cell membranes is of critical importance for osmotic balance. The current concept is that these processes are of distinct biophysical nature. Here we report the expression cloning of a liver cDNA encoding a unique promiscuous solute channel (AQP9) that confers high permeability for both solutes and water. AQP9 mediates passage of a wide variety of non-charged solutes including carbamides, polyols, purines, and pyrimidines in a phloretin- and mercury-sensitive manner, whereas amino acids, cyclic sugars, Na+, K+, Cl-, and deprotonated monocarboxylates are excluded. The properties of AQP9 define a new evolutionary branch of the major intrinsic protein family of aquaporin proteins and describe a previously unknown mechanism by which a large variety of solutes and water can pass through a single pore, enabling rapid cellular uptake or exit of metabolites with minimal osmotic perturbation. PMID- 9733775 TI - The cation-binding domain from the alpha subunit of integrin alpha5 beta1 is a minimal domain for fibronectin recognition. AB - The cation-binding domain from the alpha subunit of human integrin alpha5beta1 was produced as a recombinant protein, alpha5-(229-448). This protein displays a well defined fold with a content of 30-35% alpha-helix and 20-25% beta-strand, based on circular dichroism. The binding of Ca2+ or Mg2+ to alpha5-(229-448) results in a biphasic conformational rearrangement consistent with the occurrence of two classes of cation-binding sites differing by their affinities. The two classes of sites are located in two conformationally independent lobes, as established by a parallel study of two recombinant half-domains (N- and C terminal) that also adopt stable folds. Upon saturation with divalent cations, alpha5-(229-448) binds an Arg-Gly-Asp (RGD)-containing fibronectin ligand to form a 1:1 complex. Complex formation is associated with a specific conformational adaptation of the ligand, suggesting an induced fit mechanism. In contrast, neither of the half-domains is competent for ligand binding. The alpha5-(229-448) fibronectin complex is dissociated in the presence of an RGD peptide, as well as of a simple carboxylic acid, suggesting that the RGD aspartyl carboxylate is an essential element that directly interacts with the alpha5 cation-binding domain. PMID- 9733776 TI - The sequence, bacterial expression, and functional reconstitution of the rat mitochondrial dicarboxylate transporter cloned via distant homologs in yeast and Caenorhabditis elegans. AB - The dicarboxylate carrier (DIC) belongs to a family of transport proteins found in the inner mitochondrial membranes. The biochemical properties of the mammalian protein have been characterized, but the protein is not abundant. It is difficult to purify and had not been sequenced. We have used the sequence of the distantly related yeast DIC to identify a related protein encoded in the genome of Caenorhabditis elegans. Then, related murine expressed sequence tags were identified with the worm sequence, and the murine sequence was used to isolate the cDNA for the rat homolog. The sequences of the worm and rat proteins have features characteristic of the family of mitochondrial transport proteins. Both proteins were expressed in bacteria and reconstituted into phospholipid vesicles where their transport characteristics closely resembled those of whole rat mitochondria and of the rat DIC reconstituted into vesicles. As expected from the role of the DIC in gluconeogenesis and ureogenesis, its transcripts were detected in rat liver and kidney, but unexpectedly, they were also detected in rat heart and brain tissues where the protein may fulfill other roles, possibly in supplying substrates to the Krebs cycle. PMID- 9733777 TI - Transcription factor MSY-1 regulates expression of the murine growth hormone receptor gene. AB - Previous studies identified and partially characterized a 42-base pair regulatory element in the 5'-flanking region of the L1 transcript of the murine growth hormone (GH) receptor gene that interacted with both double- and single-stranded DNA-binding proteins. We present evidence that the double-stranded DNA-binding protein is NF-Y, a CCAAT box-binding protein. Experiments with a dominant negative form of NF-Y indicate that NF-Y does not play a direct role in regulating the activity of the FP42 element. A cDNA clone that specifically interacts with the upper (coding) strand of the regulatory element was isolated by screening a cDNA expression library using the Southwestern technique. DNA sequencing, electrophoretic mobility shift assay, Southwestern blot analysis, and supershift EMSA confirm the identity of the single-stranded binding protein to be MSY-1, a DNA-binding protein that is evolutionary conserved from prokaryotes to eukaryotes. Mapping of single-stranded DNA configurations reveals that MSY-1 can facilitate the formation of single-stranded DNA regions in the GH receptor 5' flanking region. Transient transfection experiments support the role of MSY-1 as a repressor of GH receptor gene activation. Southwestern blot analysis indicates that the levels of nuclear MSY-1 are decreased in the livers of pregnant mice, suggesting a role for MSY-1 in the increased expression of the GH receptor during pregnancy. PMID- 9733778 TI - Molecular cloning and expression of human and mouse tyrosylprotein sulfotransferase-2 and a tyrosylprotein sulfotransferase homologue in Caenorhabditis elegans. AB - Tyrosine O-sulfation, a common post-translational modification in eukaryotes, is mediated by Golgi enzymes that catalyze the transfer of the sulfuryl group from 3'-phosphoadenosine 5'-phosphosulfate to tyrosine residues in polypeptides. We recently isolated cDNAs encoding human and mouse tyrosylprotein sulfotransferase 1 (Ouyang, Y. B., Lane, W. S., and Moore, K. L. (1998) Proc. Natl. Acad. Sci. U. S. A. 95, 2896-2901). Here we report the isolation of cDNAs encoding a second tyrosylprotein sulfotransferase (TPST), designated TPST-2. The human and mouse TPST-2 cDNAs predict type II transmembrane proteins of 377 and 376 amino acid residues, respectively. The cDNAs encode functional N-glycosylated enzymes when expressed in mammalian cells. In addition, preliminary analysis indicates that TPST-1 and TPST-2 have distinct specificities toward peptide substrates. The human TPST-2 gene is on chromosome 22q12.1, and the mouse gene is in the central region of chromosome 5. We have also identified a cDNA that encodes a TPST in the nematode Caenorhabditis elegans that maps to the right arm of chromosome III. Thus, we have identified two new members of a class of membrane-bound sulfotransferases that catalyze tyrosine O-sulfation. These enzymes may catalyze tyrosine O-sulfation of a variety of protein substrates involved in diverse physiologic functions. PMID- 9733779 TI - An intact zinc ring finger is required for tumor necrosis factor receptor associated factor-mediated nuclear factor-kappaB activation but is dispensable for c-Jun N-terminal kinase signaling. AB - The diverse biological effects of the tumor necrosis factor (TNF) receptor superfamily are believed to be mediated in part through TNF receptor-associated factors (TRAFs), a family of cytoplasmic adaptor proteins which can activate intracellular signaling pathways, including the nuclear factor-kappaB (NF-kappaB) and c-Jun N-terminal kinase (JNK) pathways. TRAFs 2, 5, and 6 strongly activate both pathways when overexpressed; however, TRAF 3 (a close homologue of TRAF 5) does not significantly activate either pathway. The current study addresses the structural basis for this difference by substituting corresponding domains of TRAF 5 into TRAF 3 and testing activation of both pathways. A small region of TRAF 5 (the first zinc finger and 10 residues of the second zinc finger) is sufficient to convert TRAF 3 into an activator of both pathways. Also, an intact zinc ring finger is required for NF-kappaB activation but not JNK activation. In agreement with this finding, TRAF 2A, a TRAF 2 splice variant with an altered ring finger, is a specific activator of JNK. These findings suggest that different domains of TRAFs may be involved in NF-kappaB and JNK signaling. Also, alternative splicing of TRAFs may represent a novel mechanism whereby TNF family receptors can mediate distinct downstream effects in different tissues. PMID- 9733780 TI - Molecular cloning and characterization of the noncatalytic subunit of the Rab3 subfamily-specific GTPase-activating protein. AB - We recently purified and characterized from rat brain a GTPase-activating protein (GAP) specific for the Rab3 small G protein subfamily implicated in Ca2+ dependent exocytosis. Rab3 GAP showed two bands with Mr of about 130,000 (p130) and 150,000 (p150) on SDS-polyacrylamide gel electrophoresis. p130, but not p150, showed the catalytic activity. Because p150 was likely the subunit of Rab3 GAP, here we cloned the cDNA of p150, determined its primary structure, and characterized it. The tissue and subcellular distribution patterns of p150 and p130 were similar, and both the proteins were enriched in the synaptic soluble fraction. p150 was co-immunoprecipitated with p130 from this fraction. Recombinant p150 formed a heterodimer with recombinant p130 as estimated by sucrose density gradient ultracentrifugation. Recombinant p150 neither showed the Rab3A GAP activity nor affected the activity of recombinant p130. When p150 and p130 were co-expressed in the cells, the subcellular localization of each protein did not change. These results indicate that p150 is the noncatalytic subunit of Rab3 GAP. PMID- 9733781 TI - Requirement for both the amino-terminal catalytic domain and a noncatalytic domain for in vivo activity of ADP-ribosylation factor GTPase-activating protein. AB - The small GTP-binding protein ADP-ribosylation factor-1 (ARF1) regulates intracellular transport by modulating the interaction of coat proteins with the Golgi complex. Coat protein association with Golgi membranes requires activated, GTP-bound ARF1, whereas GTP hydrolysis catalyzed by an ARF1-directed GTPase activating protein (GAP) deactivates ARF1 and results in coat protein dissociation. We have recently cloned a Golgi-associated ARF GAP. Overexpression of GAP was found to result in a phenotype that reflects ARF1 deactivation (Aoe, T., Cukierman, E., Lee, A., Cassel, D., Peters, P. J., and Hsu, V. W. (1997) EMBO J. 16, 7305-7316). In this study, we used this phenotype to define domains in GAP that are required for its function in vivo. As expected, mutations in the amino terminal part of GAP that were previously found to abolish ARF GAP catalytic activity in vitro abrogated ARF1 deactivation in vivo. Significantly, truncations at the carboxyl-terminal part of GAP that did not affect GAP catalytic activity in vitro also diminished ARF1 deactivation. Thus, a noncatalytic domain is required for GAP activity in vivo. This domain may be involved in the targeting of GAP to the Golgi membrane. PMID- 9733782 TI - The subunit b of the F0F1-type ATPase of the bacterium Mycoplasma pneumoniae is a lipoprotein. AB - The DNA sequence analysis of the F0F1-ATPase operon of the bacterium Mycoplasma pneumoniae predicted that the subunit b, encoded by the gene atpF, is a lipoprotein of the murein lipoprotein type of Escherichia coli. Here we experimentally verify this prediction by metabolic labeling of subunit b with [14C]palmitic acid and by in vivo interfering with the processing of the prolipoprotein form of subunit b by the antibiotic globomycin, a specific inhibitor of the signal peptidase II. Our results suggest that the subunit b of the F0F1-ATPase of M. pneumoniae is anchored at the cytoplasmic membrane by an N terminal lipid modification in addition to its transmembrane domain. The lipoprotein nature of subunit b and its proposed membrane topology seems to be characteristic for mycoplasmas, since among all sequenced bacterial atpF genes, only those from Mycoplasma gallisepticum and Mycoplasma genitalium code for a conserved lipoprotein consensus sequence. PMID- 9733783 TI - Cystatin F is a glycosylated human low molecular weight cysteine proteinase inhibitor. AB - A previously undescribed human member of the cystatin superfamily called cystatin F has been identified by expressed sequence tag sequencing in human cDNA libraries. A full-length cDNA clone was obtained from a library made from mRNA of CD34-depleted cord blood cells. The sequence of the cDNA contained an open reading frame encoding a putative 19-residue signal peptide and a mature protein of 126 amino acids with two disulfide bridges and enzyme-binding motifs homologous to those of Family 2 cystatins. Unlike other human cystatins, cystatin F has 2 additional Cys residues, indicating the presence of an extra disulfide bridge stabilizing the N-terminal region of the molecule. Recombinant cystatin F was produced in a baculovirus expression system and characterized. The mature recombinant protein processed by insect cells had an N-terminal segment 7 residues longer than that of cystatin C and displayed reversible inhibition of papain and cathepsin L (Ki = 1.1 and 0.31 nM, respectively), but not cathepsin B. Like cystatin E/M, cystatin F is a glycoprotein, carrying two N-linked carbohydrate chains at positions 36 and 88. An immunoassay for quantification of cystatin F showed that blood contains low levels of the inhibitor (0.9 ng/ml). Six B cell lines in culture secreted barely detectable amounts of cystatin F, but several T cell lines and especially one myeloid cell line secreted significant amounts of the inhibitor. Northern blot analysis revealed that the cystatin F gene is primarily expressed in peripheral blood cells and spleen. Tissue expression clearly different from that of the ubiquitous inhibitor, cystatin C, was also indicated by a high incidence of cystatin F clones in cDNA libraries from dendritic and T cells, but no clones identified by expressed sequence tag sequencing in several B cell libraries and in >600 libraries from other human tissues and cells. PMID- 9733784 TI - Phosphorylation of vitronectin by casein kinase II. Identification of the sites and their promotion of cell adhesion and spreading. AB - The cell adhesion protein vitronectin (Vn) was previously shown to be the major target in human blood for an extracellular protein kinase A, which is released from platelets upon their physiological stimulation with thrombin and also prevails as an ectoenzyme in several other types of blood cells. Because plasma Vn was shown to have only one protein kinase A phosphorylation site (Ser378) but to contain approximately 3 mol of covalently bound phosphate, and because human serum and blood cells were shown to contain also a casein kinase II (CKII) on their surface, we studied the phosphorylation of Vn by CKII attempting to find out whether such phosphorylation modulates Vn function, an acid test for its having a physiological relevance. Here we show (i) that the CKII phosphorylation of Vn has a Km of 0.5-2 microM (lower than the Vn concentration in blood, 3-6 microM), (ii) that it is targeted to Thr50 and Thr57, which are vicinal to the RGD site of Vn, and (iii) that the phosphorylation of Thr57 facilitates the phosphorylation of Thr50. The maximal stoichiometry of the CKII phosphorylation of plasma Vn was found to be low, which, in principle, could be due to its partial prephosphorylation in vivo. However, for the detection of a functional modulation, we needed a comparison between a fully phosphorylated Vn (at Thr57 and Thr50) and a nonphosphorylated Vn. Therefore, we expressed Vn in a baculovirus system and show (i) that the CKII phosphorylation of wt-Vn enhances the adhesion of bovine aorta endothelial cells; (ii) that the double mutant T50E/T57E (in which the neutral Thr residues are replaced by the negatively charged Glu residues considered analogs of Thr-P) has a significantly enhanced capacity to promote cell adhesion and to accelerate cell spreading when compared with either wild-type Vn or to the neutral T50A/T57A mutant; and (iii) that, at least in the case of bovine aorta endothelial cells, the T50E/T57E mutant exhibits an enhanced adhesion, which seems to be due to an increased affinity toward the alphav beta3 Vn receptors. PMID- 9733785 TI - The transmembrane domains of ectoapyrase (CD39) affect its enzymatic activity and quaternary structure. AB - Mammalian ectoapyrase (CD39) is an integral membrane protein with two transmembrane domains and a large extracellular region. The enzymatic activity of ectoapyrase is inhibited by most detergents used for membrane protein solubilization. In contrast, the enzymatic activities of soluble E-type ATPases, including potato tuber (Solanum tuberosum) apyrase and parasite ecto-ATPase, are not affected by detergents. Here we show that ectoapyrase is a tetramer and that detergents that reduce the activity of the enzyme promote dissociation of the tetramer to monomers. We expressed a secreted form of the ectoapyrase in COS-7 cells by fusing the signal peptide of murine CD4 with the extracellular domain of the ectoapyrase. The soluble ectoapyrase is catalytically active and its activity is not affected by detergents. Mutants of the ectoapyrase with only the NH2- or the COOH-terminal transmembrane domain are membrane-bound, and their activity is no longer affected by detergents. The enzymatic activity of all of the mutant proteins is less than that of the native enzyme. These results suggest that the proper contacts between the transmembrane domains of the monomers in the tetramer are necessary for full enzymatic activity. PMID- 9733786 TI - Fidelity and mutational specificity of uracil-initiated base excision DNA repair synthesis in human glioblastoma cell extracts. AB - The fidelity of DNA synthesis associated with uracil-initiated base excision repair was measured in human whole cell extracts. An M13mp2 lacZalpha DNA-based reversion assay was developed to assess the error frequency of DNA repair synthesis at a site-specific uracil residue. All three possible base substitution errors were detected at the uracil target causing reversion of opal codon 14 in the Escherichia coli lacZalpha gene. Using human glioblastoma U251 whole cell extracts, approximately 50% of the heteroduplex uracil-containing DNA substrate was completely repaired, as determined by the insensitivity of form I DNA reaction products to cleavage by a combined treatment of E. coli uracil-DNA glycosylase and endonuclease IV. The majority of repair occurred by the uracil initiated base excision repair pathway, since the addition of the bacteriophage PBS2 uracil-DNA glycosylase inhibitor protein to extracts significantly blocked this process. In addition, the formation of repaired form I DNA molecules occurred concurrently with limited DNA synthesis, which was largely restricted to the HinfI DNA fragment initially containing the uracil residue and specific to the uracil-containing DNA strand. Based on the reversion frequency of repaired M13mp2 DNA, the fidelity of DNA repair synthesis at the target was determined to be about one misincorporated nucleotide per 1900 repaired uracil residues. The major class of base substitutions propagated transversion mutations, which were distributed almost equally between T to G and T to A changes in the template. A similar mutation frequency was also observed using whole cell extracts from human colon adenocarcinoma LoVo cells, suggesting that mismatch repair did not interfere with the fidelity measurements. PMID- 9733788 TI - Phosphotyrosine 1173 mediates binding of the protein-tyrosine phosphatase SHP-1 to the epidermal growth factor receptor and attenuation of receptor signaling. AB - The protein-tyrosine phosphatase SHP-1 binds to and dephosphorylates the epidermal growth factor receptor (EGFR), and both SH2 domains of SHP-1 are important for this interaction (Tenev, T., Keilhack, H., Tomic, S., Stoyanov, B., Stein-Gerlach, M., Lammers, R., Krivtsov, A. V., Ullrich, A., and Bohmer, F. D. (1997) J. Biol. Chem. 272, 5966-5973). We mapped the EGFR phosphotyrosine 1173 as the major binding site for SHP-1 by a combination of phosphopeptide activation, phosphopeptide competition, and receptor YF mutant analysis. Mutational conversion of the EGFR sequence 1171-1176 AEYLRV into the high affinity SHP-1 binding sequence LEYLYL of the erythropoietin receptor (EpoR) led to a highly elevated SHP-1 binding to the mutant EGFR (EGFR1171-1176EpoR) and in turn to an enhanced dephosphorylation of the receptor. SHP-1 expression interfered with EGF dependent mitogen-activated protein kinase stimulation, and this effect was more pronounced in case of EGFR1171-1176EpoR. Reduced SHP-1 binding to the EGFR Y1173F mutant resulted in a reduced receptor dephosphorylation by coexpressed SHP-1 and less interference with EGF-dependent mitogen-activated protein kinase stimulation. The effects of receptor mutations on SHP-1 binding were, however, stronger than those on receptor dephosphorylation by SHP-1. Therefore, receptor dephosphorylation may be the result of the combined activity of receptor-bound SHP-1 and SHP-1 bound to an auxiliary docking protein. PMID- 9733787 TI - Regulation of interleukin-1beta-induced interleukin-6 gene expression in human fibroblast-like synoviocytes by p38 mitogen-activated protein kinase. AB - Involvement of p38 mitogen-activated protein (MAP) kinase in interleukin (IL)-6 gene expression of human fibroblast-like synoviocytes (FLSs) was assessed. p38 MAP kinase was constitutively expressed in human FLSs and activated in response to IL-1beta. A pyridinylimidazole compound, SB203580, inhibited p38 MAP kinase activity in vivo, since the activity of MAPKAP kinase-2 (a substrate of p38 MAP kinase) in IL-1beta-stimulated FLSs was totally suppressed by it. SB203580 concentration-dependently inhibited protein production and gene expression of IL 6 by human FLSs. The effect of SB203580 was dependent on de novo protein synthesis. SB203580 significantly reduced the stability of IL-6 mRNA without affecting the rate of IL-6 gene transcription. Here, we provide evidence that p38 MAP kinase is activated in response to IL-1beta in human FLSs and is involved in IL-6 synthesis by stabilizing IL-6 mRNA. PMID- 9733789 TI - Antibody imprint of a membrane protein surface. Phagocyte flavocytochrome b. AB - Structural features of the integral membrane protein flavocytochrome b (Cyt b) were discovered using an antibody "imprint" of the Cyt b surface. Amino acid sequences were selected from a random nonapeptide phage-display library by their affinity for the monoclonal antibody 44.1 binding site, which recognizes the native conformation of the p22 subunit of Cyt b. Transferred nuclear Overhauser effect spectroscopy and rotating frame Overhauser effect spectroscopy NMR were used to study the antibody-bound conformation of a synthetic peptide derived from phage-displayed sequences. The NMR data supported the phage-display analysis suggesting the existence of a complex epitope and allowed the modeling of the close spatial proximity of the epitope components 29TAGRF33 and 183PQVNPI188 from discontinuous regions of p22. Although these regions are separated by two putative membrane-spanning domains and are 150 residues apart in the sequence, they appear to combine to form a complex epitope on the cytosolic surface of the transmembrane protein. NMR constraints, measured from the antibody-bound conformation of a composite peptide mimetic of the Cyt b epitope, and one constraint inferred from the phage-display results, were used to demonstrate the close proximity of these two regions. This information provides a low resolution view of the tertiary structure of the native discontinuous epitope on the Cyt b surface. Given additional antibodies, such imprint analysis has the potential for producing structural constraints to help support molecular modeling of this and other low abundance or noncrystallizable proteins. PMID- 9733790 TI - Localization and characterization of two nucleotide-binding sites on the anaerobic ribonucleotide reductase from bacteriophage T4. AB - We have used 8-azidoadenosine 5'-triphosphate (8-N3ATP) to investigate the nucleotide-binding sites on the NrdD subunit of the anaerobic ribonucleotide reductase from T4 phage. Saturation studies revealed two saturable sites for this photoaffinity analog of ATP. One site exhibited half-maximal saturation at approximately 5 microM [gamma-32P]8-N3ATP, whereas the other site required 45 microM. To localize the sites of photoinsertion, photolabeled peptides from tryptic and chymotryptic digests were isolated by immobilized Al3+ affinity chromatography and high performance liquid chromatography and subjected to amino acid sequence and mass spectrometric analyses. The molecular masses of the photolabeled products of cyanogen bromide cleavage were estimated using tricine SDS-polyacrylamide gel electrophoresis. Overlapping sequence analysis localized the higher affinity site to the region corresponding to residues 289-291 and the other site to the region corresponding to residues 147-160. Site-directed mutagenesis of Cys290, a residue conserved in all known class III reductases, resulted in a protein that exhibited less than 10% of wild type enzymatic activity. These observations indicate that Cys290 may reside in or near the active site. High performance liquid chromatography analysis revealed that photoinsertion of [gamma-32P]8-N3ATP into the site corresponding to residues 147 160 was almost completely abolished when 100 microM dATP, dGTP, or dTTP was included in the photolabeling reaction mixture, whereas 100 microM ATP, GTP, CTP, or dCTP had virtually no effect. Based on these nucleotide binding properties, we conclude that this site is an allosteric site analogous to the one that has been shown to regulate substrate specificity of other ribonucleotide reductases. There was no evidence for a second allosteric nucleotide-binding site as observed in the anaerobic ribonucleotide reductase from Escherichia coli. PMID- 9733791 TI - Protein kinase A-regulated instability site in the 3'-untranslated region of lactate dehydrogenase-A subunit mRNA. AB - Expression of the lactate dehydrogenase A subunit (LDH-A) gene can be controlled by transcriptional as well as posttranscriptional mechanisms. In rat C6 glioma cells, LDH-A mRNA is stabilized by activation and synergistic interaction of protein kinases A and C. In the present study, we aimed to identify the sequence domain which determines and regulates mRNA stability/instability by protein kinase A and focused our attention on the 3'-untranslated region (3'-UTR) of LDH A mRNA. We have constructed various chimeric globin/lactate dehydrogenase (ldh) genes linked to the c-fos promoter and stably transfected them into rat C6 glioma cells. After their transfection, we determined the half-life of transcribed chimeric globin/ldh mRNAs. The results showed that at least three sequence domains within the LDH-A 3'-UTR consisting of nucleotides 1286-1351, 1453-1471, and 1471-1502 are responsible for the relatively rapid rate of LDH-A mRNA turnover in the cytoplasm. Whereas chimeric globin/ldh mRNAs containing the base sequences 1286-1351 and 1453-1471 were not stabilized by (Sp)-cAMPS, an activator of protein kinase A, instability caused by the 1471-1502 domain was significantly reversed. Additional deletion and mutational analyses demonstrated that the 3' UTR fragment consisting of the 22 bases 1478-1499 is a critical determinant for the (Sp)-cAMPS-mediated LDH-A mRNA stabilizing activity. Because of its functional characteristics, we named the 22-base region "cAMP-stabilizing region." PMID- 9733792 TI - Structure and expression of the mouse AhR nuclear translocator (mArnt) gene. AB - Aryl hydrocarbon receptor (AhR) nuclear translocator (Arnt) gene has been isolated and characterized from a mouse genomic DNA library. The gene is about 60 kilobases long and split into 22 exons. An unusual exon/intron junctional sequence was found in the 11th intron of the gene that begins with GC at its 5' end. The exon/intron arrangement of mArnt gene differs greatly from those of the other members of the same basic-helix-loop-helix/PAS family. The gene is TATA less and has several transcription start sites. The promoter region of the mArnt gene is GC-rich and contains a number of putative regulatory DNA sequences such as two GC-boxes, a cAMP-responsive element, E-box, AP-1 site, and CAAT-box. Deletion experiments revealed that all these DNA elements made substantial contributions to a high level of expression of the gene, except for the cAMP responsive element. Of all, two GC-boxes displayed the most dominant enhancing effects. It was demonstrated that there exist specific factors binding to these DNA elements in the nuclear extracts of HeLa cells. Among them, Sp1 and Sp3, and CAAT-box binding factor-A were identified to bind the GC-boxes and CAAT-box, respectively. Expression of MyoD in HeLa cells stimulated the Arnt promoter activity by binding to the E-box. PMID- 9733794 TI - CREB and its associated proteins act as survival factors for human melanoma cells. AB - cAMP response element-binding protein (CREB) and activating transcription factor 1 (ATF-1), members of the CREB/ATF family, have been implicated in cAMP- and calcium-induced transcriptional activation. We have previously demonstrated that quenching of CREB-associated proteins in metastatic melanoma cells by a dominant negative CREB (KCREB) that is mutated within its DNA-binding domain decreased their radiation resistance, and their tumorigenic and metastatic potential in nude mice. As the induction of apoptosis by diverse exogenous signals is dependent on the elevation of intracellular Ca2+, the purpose of this study was to determine the role of CREB and its associated proteins in apoptosis using KCREB. We used thapsigargin (Tg), which inhibits endoplasmic reticulum-dependent Ca2+-ATPase and thereby increases cytosolic Ca2+, to induce apoptosis. MeWo human melanoma cells were transfected with the KCREB expression vector and subsequently analyzed for their susceptibility to Tg-induced apoptosis. Here we demonstrate that expression of KCREB in MeWo cells rendered them susceptible to Tg-induced apoptosis. Tg treatment induced phosphorylation of CREB and possibly ATF-1 transcription factors. Treatment with Tg induced CRE-dependent transcription in parental cells, whereas this activation was reduced in the KCREB-transfected cells. In addition, CAT activity driven by the CRE-dependent promoter was inhibited in parental MeWo cells cotransfected with increasing concentrations of KCREB in a dose-dependent manner. We did not observe any changes in Bcl-2 or Bcl 2-related proteins (Bcl-x, Bax, and Bad) in control or KCREB-transfected cells before or after treatment with Tg. Collectively, these data indicate that CREB and its associated proteins act as survival factors for human melanoma cells, and hence contribute to the acquisition of the malignant phenotype. PMID- 9733793 TI - The catalytic subunit of the cAMP-dependent protein kinase of ovine sperm flagella has a unique amino-terminal sequence. AB - The basis for the unusual properties of the catalytic subunit (C) of ram sperm cAMP-dependent protein kinase was investigated. Ram sperm C was purified and found by mass spectrometry (MS) to be approximately 890 Da smaller than Calpha, the predominant somatic isoform. Partial internal amino acid sequence from ram sperm C was an exact match to that of bovine Calpha, but differed from the predicted sequences for the Cbeta and Cgamma isoforms. MS analysis of 2-nitro-5 thiocyanatobenzoic acid fragments showed that the mass difference originated in the amino-terminal region. A unique blocked amino-terminal fragment was isolated from sperm C and sequenced by a combination of tandem mass spectrometry and Edman degradation of a subfragment. The results revealed that the amino-terminal myristate and the first 14 amino acids of Calpha are replaced by an amino terminal acetate and six different amino acids in sperm C. The predicted mass difference due to these changes is 899 Da. The region of homology between sperm C and Calpha begins at the exon 1/exon 2 boundary in Calpha, suggesting that sperm C results from use of an alternate exon 1 in the Calpha gene. The different amino terminus of sperm C may be related to a unique requirement for localization of the "free" C subunit within the sperm flagellum. PMID- 9733795 TI - Identification of a novel 81-kDa component of the Xenopus origin recognition complex. AB - The Xenopus origin recognition complex is essential for chromosomal DNA replication in cell-free extracts. We have immunopurified the Xenopus origin recognition complex with anti-Xorc2 antibodies and analyzed its composition and properties. Xorc2 (p63) is specifically associated with Xorc1 (p115) and up to four additional polypeptides (p81, p78, p45, and p40). The cDNA encoding p81 is highly homologous to various expressed sequence tags from humans and mice encoding a protein of previously unknown function. Immunodepletion of p81 from Xenopus egg extracts, which also results in the removal of Xorc2, completely abolishes chromosomal DNA replication. Thus, p81 appears to play a crucial role at S phase in higher eukaryotes. PMID- 9733796 TI - Activation of integrated provirus requires histone acetyltransferase. p300 and P/CAF are coactivators for HIV-1 Tat. AB - A unique aspect of the retrovirus life cycle is the obligatory integration of the provirus into host cell chromosomes. Unlike viruses that do not integrate, retroviruses must conserve an ability to activate transcription from a chromatin context. Human immunodeficiency virus (HIV)-1 encodes an unusual and an unusually potent transcriptional transactivator, Tat, which binds to a nascent viral leader RNA, TAR. The action of Tat has been well studied in various reductive model systems; however, the physiological mechanism through which Tat gains access to chromatin-associated proviral long terminal repeats (LTRs) is not understood. We show here that a nuclear histone acetyltransferase activity associates with Tat. Intracellularly, we found that Tat forms a ternary complex with p300 and P/CAF, two histone acetyltransferases (HATs). A murine cell defect in Tat transactivation of the HIV-1 LTR was linked to the reduced abundance of p300 and P/CAF. Thus, overexpression of p300 and P/CAF reconstituted Tat transactivation of the HIV-1 LTR in NIH3T3 cells to a level similar to that observed for human cells. By using transdominant p300 or P/CAF mutants that lack enzymatic activity, we delineated a requirement for the HAT component from the latter but not the former in Tat function. Finally, we observed that Tat-associated HAT is preferentially important for transactivation of integrated, but not unintegrated, HIV-1 LTR. PMID- 9733797 TI - Requirements for and regulation of origin opening of plasmid P1. AB - Origin opening is essential for the initiation of DNA replication in the theta mode and requires binding of initiator proteins. Using reactivity to KMnO4 in vivo as an assay, we find that, like initiation, origin opening of the Escherichia coli plasmid P1 requires the host initiators DnaA and HU and the plasmid-encoded initiator RepA. The ability to detect opening at the P1ori in vivo allowed us to study this activity at various copy numbers in chimeric replicons. The opening was prevented when the P1ori was cloned in high copy vectors or when excess RepA binding sites (iterons) were provided in trans. However, when RepA supply was also increased, the opening was efficient. A further increase in RepA prevented opening. Replication of an incoming P1 under these conditions correlated with opening. These results demonstrate that initiation is possible even at abnormally high origin concentrations and that oversupply of RepA, relative to iterons, can prevent replication by blocking origin opening. It appears that plasmid overreplication can be prevented either by limiting RepA or by accumulating RepA at a rate higher than that of the origin. PMID- 9733798 TI - Mutations in and monoclonal antibody binding to evolutionary hypervariable region of Escherichia coli RNA polymerase beta' subunit inhibit transcript cleavage and transcript elongation. AB - A 190 amino acid-long region centered around position 1050 of the 1407-amino acid long beta' subunit of Escherichia coli RNA polymerase (RNAP) is absent from homologues in eukaryotes, archaea and many bacteria. In chloroplasts, the corresponding region can be more than 900 amino acids long. The role of this hypervariable region was studied by deletion mutagenesis of the cloned E. coli rpoC, encoding beta'. Long deletions mimicking beta' from Gram-positive bacteria failed to assemble into RNAP. Mutants with short, 40-60-amino acid-long deletions spanning beta' residues 941-1130 assembled into active RNAP in vitro. These mutant enzymes were defective in the transcript cleavage reaction and had dramatically reduced transcription elongation rates at subsaturating substrate concentrations due to prolonged pausing at sites of transcriptional arrest. Binding of a monoclonal antibody, Pyn1, to the hypervariable region inhibited transcription elongation and intrinsic transcript cleavage and, to a lesser degree, GreB-induced transcript cleavage, but did not interfere with GreB binding to RNAP. We propose that mutations in and antibody binding to the hypervariable, functionally dispensable region of beta' inhibit transcript cleavage and elongation by distorting the flanking conserved segment G in the active center. PMID- 9733799 TI - Membrane integration of Na,K-ATPase alpha-subunits and beta-subunit assembly. AB - The control of membrane insertion of polytopic proteins is still poorly understood. We carried out in vivo translation/insertion experiments in Xenopus oocytes with combined wild type or mutant membrane segments of the alpha-subunit of the heterodimeric Na, K-ATPase linked to a glycosylation reporter sequence. We confirm that the four N-terminal hydrophobic segments of the alpha-subunit behave as alternating signal anchor/stop transfer motifs necessary for two lipid inserted membrane pairs. For the six C-terminal membrane segments, however, proper packing depends on specific sequence information and association with the beta-subunit. M5 is a very inefficient signal anchor sequence due to the presence of prolines and polar amino acids. Its correct membrane insertion is probably mediated by posttranslational hairpin formation with M6, which is favored by a proline pair in the connecting loop. M7 has partial signal anchor function, which may be mediated by the presence of glycine and glutamine residues. The formation of a transmembrane M7/M8 pair requires the association of the beta-subunit, which induces a conformational change in the connecting extracytoplasmic loop that favors M7/M8 packing. The formation of the M9/M10 pair appears to be predominantly mediated by the efficient stop transfer function of M10. Mutations that provide signal anchor function to M5, M7, and M9 abolish or impede the transport activity of the enzyme. These data illustrate the importance of specific amino acids near or within hydrophobic regions as well as of subunit oligomerization for correct topographical alignment that is necessary for proper folding and/or activity of oligomeric membrane proteins. PMID- 9733800 TI - Platelet-derived growth factor-specific regulation of the JE promoter in rat aortic smooth muscle cells. AB - JE is a member of the family of "immediate early" genes induced by growth factors and cytokines. JE encodes a low molecular weight secretory glycoprotein analogous to the human monocyte chemoattractant protein, MCP-1. JE and MCP-1 proteins are thought to play an important role in inflammation and in the recruitment of monocyte/macrophages to the vessel wall during the development of atherosclerosis. We have previously reported that the induction of JE in rat aortic smooth muscle cells (SMC) was specific to platelet-derived growth factor (PDGF) and was not seen with other growth agonists. Using a luciferase reporter system and transient transfection assays of rat aortic SMC, we now report the identification of a region in the proximal rat JE promoter that is responsive to PDGF but not to other growth factors (angiotensin II and alpha-thrombin) or cytokines (interleukin 1-beta and tumor necrosis factor-alpha). The full response to PDGF (approximately 6-fold) requires the cooperative activity of two potentially novel cis-acting elements, at positions -146 to -128 and -84 to -59. While each element produces a different pattern in electrophoretic mobility shift assays, they appear to bind the same PDGF-responsive species. Further analysis of these regions should provide important insights into PDGF-specific responses in vascular SMC. PMID- 9733801 TI - Modulation of kinase activity and oncogenic properties by alternative splicing reveals a novel regulatory mechanism for B-Raf. AB - Members of the raf oncogene family encode serine/threonine protein kinases, which activate the mitogen-activated protein kinase kinase MEKs (MAPK or ERK kinases) through direct interaction and phosphorylation. Several recent studies have revealed interesting differences between two members of this family, Raf-1 and B Raf, regarding their activation, regulation, and kinase activity. In particular, B-Raf was shown to display higher MEK kinase activity than Raf-1. By using both two-hybrid analysis and coimmunoprecipitation experiments, we demonstrate here that B-Raf also markedly differs from Raf-1 by a higher affinity for MEK. We previously reported that the B-raf gene encodes multiple protein isoforms resulting from complex alternative splicing of two exons (exons 8b and 10) located upstream of B-Raf kinase domain. In the present study, we show that these naturally occurring modifications within the protein sequence markedly modulate both the biochemical and oncogenic properties of B-Raf. The presence of exon 10 sequences enhances the affinity for MEK, the basal kinase activity, as well as the mitogenic and transforming properties of full-length B-Raf, whereas the presence of exon 8b sequences seems to have opposite effects. Therefore, alternative splicing represents a novel regulatory mechanism for a protein of the Raf family. PMID- 9733802 TI - Identification and cloning of a glucan- and lipopolysaccharide-binding protein from Eisenia foetida earthworm involved in the activation of prophenoloxidase cascade. AB - Coelomic fluid of Eisenia foetida earthworms contains a 42-kDa protein named coelomic cytolytic factor 1 (CCF-1) that was described previously to be involved in cytolytic, opsonizing, and hemolytic properties of the coelomic fluid. Cloning and sequencing of CCF-1 reveal significant homology with the putative catalytic region of beta-1,3- and beta-1,3-1,4-glucanases. CCF-1 also displays homology with coagulation factor G from Limulus polyphemus and with Gram-negative bacteria binding protein of Bombyx mori silkworm, two proteins involved in invertebrate defense mechanisms. We show that CCF-1 efficiently binds both beta-1,3-glucan and lipopolysaccharide. Moreover, CCF-1 participates in the activation of prophenoloxidase cascade via recognition of yeast and Gram-negative bacteria cell wall components. These results suggest that the 42-kDa CCF-1 protein of E. foetida coelomic fluid likely plays a role in the protection of earthworms against microbes. PMID- 9733803 TI - Pro-tumor necrosis factor-alpha processing activity is tightly controlled by a component that does not affect notch processing. AB - The extracellular domain of a heterogeneous group of transmembrane proteins can be proteolytically released from the cell surface, a process known as protein ectodomain shedding. Despite the biomedical importance of several substrates of the shedding system, such as the beta-amyloid precursor protein (betaAPP), little is known about the regulation of protein ectodomain shedding, and the only protease known to be involved is the metalloprotease disintegrin, tumor necrosis factor-alpha converting enzyme (TACE). Here, we show that previously described pro-transforming growth factor-alpha shedding-defective cell mutants (M2 cells), known to be defective in ectodomain shedding of several molecules, that include betaAPP, fail to shed the ectodomain of pro-TNF-alpha. The target of the mutation is a component required for TACE activity, since transfection of TACE into M2 cells has no effect on the shedding of pro-TNF-alpha and somatic cell fusions between M2 cells and TACE null cells recover the ability to shed pro-TNF-alpha, pro-transforming growth factor-alpha, and betaAPP. Furthermore, we show that TACE is also necessary for the shedding of betaAPP since TACE null cells show defective betaAPP shedding. Biochemical evidence shows that the component that controls TACE is different from protein kinase C, the only known activator of protein ectodomain shedding, and that this component does not affect biosynthesis or processing of TACE or other metalloprotease disintegrins. The component mutated in M2 cells is likely to control only a subset of metalloprotease disintegrins involved in regulated ectodomain shedding, since Notch processing, a process known to be dependent on the activity of another metalloprotease disintegrin, Kuzbanian, is normal in M2 cells. PMID- 9733804 TI - Transmembrane protein insertion orientation in yeast depends on the charge difference across transmembrane segments, their total hydrophobicity, and its distribution. AB - The determinants of transmembrane protein insertion orientation at the endoplasmic reticulum have been investigated in Saccharomyces cerevisiae using variants of a Type III (naturally exofacial N terminus (Nexo)) transmembrane fusion protein derived from the N terminus of Ste2p, the alpha-factor receptor. Small positive and negative charges adjacent to the transmembrane segment had equal and opposite effects on orientation, and this effect was independent of N- or C-terminal location, consistent with a purely electrostatic interaction with response mechanisms. A 3:1 bias toward Nexo insertion, observed in the absence of a charge difference, was shown to reflect the Nexo bias conferred by longer transmembrane segments. Orientation correlated best with total hydrophobicity rather than length, but it was also strongly affected by the distribution of hydrophobicity within the transmembrane segment. The most hydrophobic terminus was preferentially translocated. Insertion orientation thus depends on integration of responses to at least three parameters: charge difference across a transmembrane segment, its total hydrophobicity, and its hydrophobicity gradient. Relative signal strengths were estimated, and consequences for topology prediction are discussed. Responses to transmembrane sequence may depend on protein-translocon interactions, but responses to charge difference may be mediated by the electrostatic field provided by anionic phospholipids. PMID- 9733805 TI - Adenine nucleoside 3'-tetraphosphates are novel and potent inhibitors of adenylyl cyclases. AB - 2'-Deoxyadenosine 3'-tetraphosphate (2'-deoxy-3'-A4P) and 2', 5'-dideoxyadenosine 3'-tetraphosphate (2',5'-dideoxy-3'-A4P) were synthesized, and their effects were tested on crude and purified forms of native adenylyl cyclases isolated from brain. Syntheses combined the method of alkoxide activation with the use of tribromoethyl phosphoromorpholino-chloridate as an initial phosphorylating agent. Inhibition of adenylyl cyclase was rapid in onset. With 2'-d-3'-A4P or 2',5'-dd 3'-A4P inhibition of a purified native enzyme conformed to a linear noncompetitive behavior with respect to substrate, metal-5'ATP. Order of potency was 2', 5'-dideoxy- > 2'-deoxyadenosine and 3'-tetraphosphate > 3'-triphosphate. Both mechanism of inhibition and rank order of potency were consistent with inhibition via the 3'-nucleotide-(P)-site on adenylyl cyclase. Neither 2',5'-dd 3'-ATP nor 2',5'-dd-3'-A4P had any effect on the activities of other adenosine nucleotide binding proteins such as Ca2+/calmodulin-sensitive cyclic nucleotide phosphodiesterase, Na+/K+-ATPase, or cAMP-dependent protein kinase. With purified adenylyl cyclase from bovine brain 2',5'-dd-3'-A4P and 2'-d-3'-A4P gave, respectively, IC50 values of 9.3 and 15 nM and Ki values of 23 and 53 nM. These 3'-nucleotides are the most potent regulators described for adenylyl cyclases. PMID- 9733806 TI - The role of herpes simplex virus ICP27 in the regulation of UL24 gene expression by differential polyadenylation. AB - Herpes simplex virus specifies two sets of transcripts from the UL24 gene, short transcripts (e.g., 1.4 kb), processed at the UL24 poly(A) site, and long transcripts (e.g., 5.6 kb), processed at the UL26 poly(A) site. The 1.4- and 5.6 kb transcripts initiate from the same promoter but are expressed with early and late kinetics, respectively. Measurements of transcript levels following actinomycin D treatment of infected cells revealed that the 1.4- and 5.6-kb UL24 transcripts have similar stabilities, consistent with UL24 transcript kinetics being regulated by differential polyadenylation rather than by differential stabilities. Although the UL24 poly(A) site, which gives rise to short transcripts, is encountered first during processing, long transcripts processed at the UL26 site are equally or more abundant; thus, operationally, the UL24 site is weak. Using a series of viral ICP27 mutants, we investigated whether ICP27, which has been suggested to stimulate the usage of weak poly(A) sites, stimulates 1.4-kb transcript accumulation. We found that accumulation of 1.4-kb transcripts did not require ICP27 during viral infection. Rather, ICP27 was required for full expression of 5.6-kb transcripts, and the decrease in 5. 6-kb transcripts relative to 1.4-kb transcripts was not due solely to reduced DNA synthesis. Our results indicate that temporal expression of UL24 transcripts can be regulated by differential polyadenylation and that although ICP27 is not required for processing at the operationally weak UL24 poly(A) site, it does modulate 5.6-kb transcript levels at a step subsequent to transcriptional initiation. PMID- 9733807 TI - Local periocular vaccination protects against eye disease more effectively than systemic vaccination following primary ocular herpes simplex virus infection in rabbits. AB - Vaccination of experimental animals can provide efficient protection against ocular herpes simplex virus type 1 (HSV-1) challenge. Although it is suspected that local immune responses are important in protection against ocular HSV-1 infection, no definitive studies have been done to determine if local ocular vaccination would produce more efficacious protection against HSV-1 ocular challenge than systemic vaccination. To address this question, we vaccinated groups of rabbits either systemically or periocularly with recombinant HSV-2 glycoproteins B (gB2) and D (gD2) in MF59 emulsion or with live KOS (a nonneurovirulent strain of HSV-1). Three weeks after the final vaccination, all eyes were challenged with McKrae (a virulent, eye disease-producing strain of HSV 1). Systemic vaccination with either HSV-1 KOS or gB2/gD2 in MF59 did not provide significant protection against any of the four eye disease parameters measured (conjunctivitis, iritis, epithelial keratitis, and corneal clouding). In contrast, periocular vaccination with gB2/gD2 in MF59 provided significant protection against conjunctivitis and iritis, while ocular vaccination with live HSV-1 KOS provided significant protection against all four parameters. Thus, local ocular vaccination provided better protection than systemic vaccination against eye disease following ocular HSV-1 infection. Since local vaccination should produce a stronger local immune response than systemic vaccination, these results suggest that the local ocular immune response is very important in protecting against eye disease due to primary HSV-1 infection. Thus, for clinical protection against primary HSV-1-induced corneal disease, a local ocular vaccine may prove more effective than systemic vaccination. PMID- 9733808 TI - Cytomegalovirus assembly protein precursor and proteinase precursor contain two nuclear localization signals that mediate their own nuclear translocation and that of the major capsid protein. AB - The cytomegalovirus (CMV) assembly protein precursor (pAP) interacts with the major capsid protein (MCP), and this interaction is required for nuclear translocation of the MCP, which otherwise remains in the cytoplasm of transfected cells (L. J. Wood et al., J. Virol. 71:179-190, 1997). We have interpreted this finding to indicate that the CMV MCP lacks its own nuclear localization signal (NLS) and utilizes the pAP as an NLS-bearing escort into the nucleus. The CMV pAP amino acid sequence has two clusters of basic residues (e.g., KRRRER [NLS1] and KARKRLK [NLS2], for simian CMV) that resemble the simian virus 40 large-T-antigen NLS (D. Kalderon et al., Cell 39:499-509, 1984) and one of these (NLS1) has a counterpart in the pAP homologs of other herpesviruses. The work described here establishes that NLS1 and NLS2 are mutually independent NLS that can act (i) in cis to translocate pAP and the related proteinase precursor (pNP1) into the nucleus and (ii) in trans to transport MCP into the nucleus. By using combinations of NLS mutants and carboxy-terminal deletion constructs, we demonstrated a self-interaction of pAP and cytoplasmic interactions of pAP with pNP1 and of pNP1 with itself. The relevance of these findings to early steps in capsid assembly, the mechanism of MCP nuclear transport, and the possible cytoplasmic formation of protocapsomeric substructures is discussed. PMID- 9733809 TI - Control of cytomegalovirus in bone marrow transplantation chimeras lacking the prevailing antigen-presenting molecule in recipient tissues rests primarily on recipient-derived CD8 T cells. AB - Cytomegalovirus (CMV) infection during the transient immunodeficiency after bone marrow transplantation (BMT) develops into disease unless antiviral CD8 T cells are restored in due course. Histoincompatibility between donor and recipient is associated with increased risk. Complications may include a rejection response against the foreign major histocompatibility complex (MHC) antigens and a lack of antiviral control resulting from a misfit between donor-derived T cells and the antigenic viral peptides presented in recipient tissues. Here we have established a murine model of CMV disease after experimental BMT performed across a single MHC class I disparity. Specifically, BALB/c bone marrow cells expressing the prevailing antigen-presenting molecule Ld were transplanted into the Ld gene deletion mutant BALB/c-H-2(dm2), an experimental setting that entails a selective risk of host-versus-graft but not graft-versus-host response. The reconstituted T cell population proved to be chimeric in that it consisted of Ld-positive donor derived and Ld-negative recipient-derived cells. Pulmonary infiltrates did not include cytolytic T cells directed against Ld. This finding implies that the infection did not trigger a host-versus-graft response. Notably, upon adoptive transfer, donor-derived CD8 T cells preferentially protected tissues of donor genotype, whereas recipient-derived CD8 T cells protected tissues of either genotype. We infer from these data that the focus on immunodominant antigens presented by Ld within the donor cell population distracted the donor T cells from protecting recipient tissues and that protection in the chimeras was therefore primarily based on recipient T cells. As a consequence, T-cell chimerism after BMT should give a positive prognosis with respect to control of CMV. PMID- 9733811 TI - Recombinant measles viruses with mutations in the C, V, or F gene have altered growth phenotypes in vivo. AB - An understanding of the determinants of measles virus (MV) virulence has been hampered by the lack of an experimental model of infection. We have previously demonstrated that virulence phenotypes in human infections are faithfully reproduced by infection of human thymus/liver (thy/liv) implants engrafted into SCID mice, where the virus grows primarily in stromal cells but induces thymocyte apoptosis (P. G. Auwaerter et al., J. Virol. 70:3734-3740, 1996). To begin to elucidate the roles of the C protein, V protein, and the 5' untranslated region of the F gene (F 5'UTR) in MV infection in vivo, the replication of strains bearing mutations of these genes was compared to that of the parent sequence tagged Edmonston strain (EdTag). Growth curves show that mutants fall into two phenotypic classes. One class of mutants demonstrated kinetics of growth similar to that of EdTag, with decreased peak titers. The second class of mutants manifested peak titers similar to that of EdTag but had different replication kinetics. Abrogation of V expression led to delayed and markedly prolonged replication. Additionally, thymocyte survival was prolonged and implant architecture was preserved throughout the course of infection. In contrast, massive bystander thymocyte death occurred after infection with EdTag and all other mutants. A mutant which overexpressed V in Vero cells (V+) had the opposite phenotype of the A mutant not expressing V (V-). V+ grew more rapidly than EdTag with 100-fold-greater levels of virus production 3 days after infection. These results suggest that C, V, and the F 5'UTR are accessory factors required for efficient virus replication in vivo. In addition, thymocyte survival after V- infection suggests this protein may play multiple roles in pathogenesis of MV infection of thymus. Since these recombinant mutant viruses grew identically to the parent virus in Vero cells, the data show that thy/liv implants are an excellent model for investigating the determinants of MV virulence. PMID- 9733810 TI - Membrane fusion promoted by increasing surface densities of the paramyxovirus F and HN proteins: comparison of fusion reactions mediated by simian virus 5 F, human parainfluenza virus type 3 F, and influenza virus HA. AB - The membrane fusion reaction promoted by the paramyxovirus simian virus 5 (SV5) and human parainfluenza virus type 3 (HPIV-3) fusion (F) proteins and hemagglutinin-neuraminidase (HN) proteins was characterized when the surface densities of F and HN were varied. Using a quantitative content mixing assay, it was found that the extent of SV5 F-mediated fusion was dependent on the surface density of the SV5 F protein but independent of the density of SV5 HN protein, indicating that HN serves only a binding function in the reaction. However, the extent of HPIV-3 F protein promoted fusion reaction was found to be dependent on surface density of HPIV-3 HN protein, suggesting that the HPIV-3 HN protein is a direct participant in the fusion reaction. Analysis of the kinetics of lipid mixing demonstrated that both initial rates and final extents of fusion increased with rising SV5 F protein surface densities, suggesting that multiple fusion pores can be active during SV5 F protein-promoted membrane fusion. Initial rates and extent of lipid mixing were also found to increase with increasing influenza virus hemagglutinin protein surface density, suggesting parallels between the mechanism of fusion promoted by these two viral fusion proteins. PMID- 9733812 TI - Characterization of and functional antigen presentation by central nervous system mononuclear cells from mice infected with Theiler's murine encephalomyelitis virus. AB - We examined the phenotype and function of cells infiltrating the central nervous system (CNS) of mice persistently infected with Theiler's murine encephalomyelitis virus (TMEV) for evidence that viral antigens are presented to T cells within the CNS. Expression of major histocompatibility complex (MHC) class II in the spinal cords of mice infected with TMEV was found predominantly on macrophages in demyelinating lesions. The distribution of I-As staining overlapped that of the macrophage marker sialoadhesin in frozen sections and coincided with that of another macrophage/microglial cell marker, F4/80, by flow cytometry. In contrast, astrocytes, identified by staining with glial fibrillary acidic protein, rarely expressed detectable MHC class II, although fibrillary gliosis associated with the CNS damage was clearly seen. The costimulatory molecules B7-1 and B7-2 were expressed on the surface of most MHC class II positive cells in the CNS, at levels exceeding those found in the spleens of the infected mice. Immunohistochemistry revealed that B7-1 and B7-2 colocalized on large F4/80(+) macrophages/microglia in the spinal cord lesions. In contrast, CD4(+) T cells in the lesions expressed mainly B7-2, which was found primarily on blastoid CD4(+) T cells located toward the periphery of the lesions. Most interestingly, plastic-adherent cells freshly isolated from the spinal cords of TMEV-infected mice were able to process and present TMEV and horse myoglobin to antigen-specific T-cell lines. Furthermore, these cells were able to activate a TMEV epitope-specific T-cell line in the absence of added antigen, providing conclusive evidence for the endogenous processing and presentation of virus epitopes within the CNS of persistently infected SJL/J mice. PMID- 9733813 TI - Effect of immune activation on the dynamics of human immunodeficiency virus replication and on the distribution of viral quasispecies. AB - Virus replication in a human immunodeficiency virus (HIV)-infected individual, as determined by the steady-state level of plasma viremia, reflects a complex balance of viral and host factors. We have previously demonstrated that immunization of HIV-infected individuals with the common recall antigen, tetanus toxoid, disrupts this steady state, resulting in transient bursts of plasma viremia after immunization. The present study defines the viral genetic basis for the transient bursts in viremia after immune activation. Tetanus immunization was associated with dramatic and generally reversible shifts in the composition of plasma viral quasispecies. The viral bursts in most cases reflected a nonspecific increase in viral replication secondary to an expanded pool of susceptible CD4(+) T cells. An exception to this was in a patient who harbored viruses of differing tropisms (syncytium inducing and non-syncytium inducing [NSI]). In this situation, immunization appeared to select for the replication of NSI viruses. In one of three patients, the data suggested that immune activation resulted in the appearance in plasma of virus induced from latently infected cells. These findings illustrate certain mechanisms whereby antigenic stimulation may influence the dynamics of HIV replication, including the relative expression of different viral variants. PMID- 9733814 TI - The NH2 terminus of the herpes simplex virus type 1 regulatory protein ICP0 contains a promoter-specific transcription activation domain. AB - The transcriptional program of herpes simplex virus is regulated by the concerted action of three immediate-early (alpha) proteins, ICP4, ICP27, and ICP0. The experiments described in this study examine the role of the acidic amino terminus (amino acids 1 to 103) of ICP0 in gene activation. When tethered to a DNA binding domain, this sequence activates transcription in the yeast Saccharomyces cerevisiae. Deletion of these amino acids affects the ability of ICP0 to activate alpha-gene promoter reporters in transient expression assays, while it has little or no effect on a beta- and a gamma-gene reporter in the same assay. Viruses that express the deleted form of ICP0 (ICP0-NX) have a small-plaque phenotype on both Vero cells and the complementing cell line L7. Transient expression and immunofluorescence analyses demonstrate that ICP0-NX is a dominant negative form of ICP0. Immunoprecipitation of ICP0 from cells coinfected with viruses expressing ICP0-NX and ICP0 revealed that ICP0 oligomerizes in infected cells. These data, in conjunction with the finding that ICP0-N/X is dominant negative, provide both biochemical and genetic evidence that ICP0 functions as a multimer in infected cells. PMID- 9733815 TI - Tyrosine 112 of latent membrane protein 2A is essential for protein tyrosine kinase loading and regulation of Epstein-Barr virus latency. AB - Latent membrane protein 2A (LMP2A) of Epstein-Barr virus (EBV) is expressed on the plasma membrane of B lymphocytes latently infected with EBV and blocks B-cell receptor (BCR) signal transduction in EBV-immortalized B cells in vitro. The LMP2A amino-terminal domain that is essential for the LMP2A-mediated block on BCR signal transduction contains eight tyrosine residues. Association of Syk protein tyrosine kinase (PTK) with LMP2A occurs at the two tyrosines of the LMP2A immunoreceptor tyrosine-based activation motif, and it is hypothesized that Lyn PTK associates with the YEEA amino acid motif at LMP2A tyrosine 112 (Y112). To examine the specific association of Lyn PTK to LMP2A, a panel of LMP2A cDNA expression vectors containing LMP2A mutations were transfected into an EBV negative B-cell line and analyzed for Lyn and LMP2A coimmunoprecipitation. Lyn associates with wild-type LMP2A and other LMP2A mutant constructs, but Lyn association is lost in the LMP2A construct containing a tyrosine (Y)-to phenylalanine (F) mutation at LMP2A residue Y112 (LMP2AY112F). Next, the LMP2AY112F mutation was recombined into the EBV genome to generate stable lymphoblastoid cell lines (LCLs) transformed with the LMP2AY112F mutant virus. Analysis of BCR-mediated signal transduction in the LMP2AY112F LCLs revealed loss of the LMP2A-mediated block in BCR signal transduction. In addition, LMP2A was not tyrosine phosphorylated in LMP2AY112F LCLs. Together these data indicate the importance of the LMP2A Y112 residue in the ability of LMP2A to block BCR mediated signal transduction and place the role of this residue and its interaction with Lyn PTK as essential to LMP2A phosphorylation, PTK loading, and down-modulation of PTKs involved in BCR-mediated signal transduction. PMID- 9733816 TI - The secondary structure of the R region of a murine leukemia virus is important for stimulation of long terminal repeat-driven gene expression. AB - In addition to their role in reverse transcription, the R-region sequences of some retroviruses affect viral transcription. The first 28 nucleotides of the R region within the long terminal repeat (LTR) of the murine type C retrovirus SL3 were predicted to form a stem-loop structure. We tested whether this structure affected the transcriptional activity of the viral LTR. Mutations that altered either side of the stem and thus disrupted base pairing were generated. These decreased the level of expression of a reporter gene under the control of viral LTR sequences about 5-fold in transient expression assays and 10-fold in cells stably transformed with the LTR-reporter plasmids. We also generated a compensatory mutant in which both the ascending and descending sides of the stem were mutated such that the nucleotide sequence was different but the predicted secondary structure was maintained. Most of the activity of the wild-type SL3 element was restored in this mutant. Thus, the stem-loop structure was important for the maximum activity of the SL3 LTR. Primer extension analysis indicated that the stem-loop structure affected the levels of cytoplasmic RNA. Nuclear run-on assays indicated that deletion of the R region had a small effect on transcriptional initiation and no effect on RNA polymerase processivity. Thus, the main effect of the R-region element was on one or more steps that occurred after the template was transcribed by RNA polymerase. This finding implied that the main function of the R-region element involved RNA processing. R-region sequences of human immunodeficiency virus type 1 or mouse mammary tumor virus could not replace the SL3 element. R-region sequences from an avian reticuloendotheliosis virus partially substituted for the SL3 sequences. R-region sequences from Moloney murine leukemia virus or feline leukemia virus did function in place of the SL3 element. Thus, the R region element appears to be a general feature of the mammalian type C genus of retroviruses. PMID- 9733817 TI - Apoptotic regulation of T cells and absence of immune deficiency in virus infected gamma interferon receptor knockout mice. AB - Acute viral infections often induce a transient period of immune deficiency in which the host's T cells fail to proliferate in response to T-cell mitogens and fail to make an antigen-specific memory recall response. This has been associated with the enhanced sensitivity of these highly activated T cells to undergo apoptosis, or activation-induced cell death (AICD), upon T-cell receptor ligation. Here we show that gamma interferon receptor-deficient (IFN-gamma R-/-) mice mount a T-cell response to lymphocytic choriomeningitis virus (LCMV) infection but fail to undergo the transient immune deficiency. Instead, their T cells were hyperproliferative and relatively, but not completely, resistant to AICD. The immune response returned to homeostasis, but with delayed kinetics, in parallel with delayed clearance of the virus. Wild-type mice receiving high doses of disseminating LCMV Clone 13 are known to undergo clonal exhaustion of their virus-specific cytotoxic T lymphocytes (CTL). To determine whether this process was mediated by AICD associated with IFN-gamma or with Fas-Fas ligand interactions, LCMV-specific precursor CTL frequencies were examined in LCMV Clone 13-infected IFN-gamma R-/- or lpr (Fas-deficient) mice. In both instances, viral persistence was established and CTL precursors were greatly eliminated. This finding indicates that clonal exhaustion of CTL does not require IFN-gamma or Fas, even though both molecules influence AICD and the transient immune deficiency seen in the LCMV infection. PMID- 9733818 TI - Dendritic cells route human immunodeficiency virus to lymph nodes after vaginal or intravenous administration to mice. AB - We have developed a murine model to study the involvement of dendritic cells (DC) in human immunodeficiency virus (HIV) routing from an inoculation site to the lymph nodes (LN). Murine bone marrow-derived DC migrate to the draining LN within 24 h after subcutaneous injection. After incubation of these cells with heat inactivated (Hi) HIV type 1 (HIV-1), HIV RNA sequences were detected in the draining LN only. Upon injection of DC pulsed with infectious HIV, the virus recovered in the draining LN was still able to productively infect human T cells. After a vaginal challenge with Hi HIV-1, the virus could be detected in the iliac and sacral draining LN at 24 h after injection. After an intravenous challenge, the virus could be detected in peripheral LN as soon as 30 min after injection. The specific depletion of a myeloid-related LN DC population, previously shown to take up blood macromolecules and to translocate them into the LN, prevented HIV transport to LN. Together, our data demonstrate the critical role of DC for HIV routing to LN after either a vaginal or an intravenous challenge, which does not require their infection. Therefore, despite the fact that the mouse is not infectable by HIV, this small animal model might be useful to test preventive strategies against HIV. PMID- 9733819 TI - Serp2, an inhibitor of the interleukin-1beta-converting enzyme, is critical in the pathobiology of myxoma virus. AB - Recently, myxoma virus was shown to encode an additional member of the serpin superfamily. The viral gene, called serp2, was cloned, and the Serp2 protein was shown to specifically bind to interleukin-1beta (IL-1beta)-converting enzyme (ICE), thus inhibiting the cleavage of pro-IL-1beta by the protease (F. Petit, S. Bertagnoli, J. Gelfi, F. Fassy, C. Boucraut-Baralon, and A. Milon, J. Virol. 70:5860-5866, 1996). Here, we address the role of Serp2 in the development of myxomatosis, a lethal infectious disease of the European rabbit. A Serp2 mutant myxoma virus was constructed by disruption of the single-copy serp2 gene and insertion of the Escherichia coli gpt gene serving as the selectable marker. A revertant virus was obtained by replacing the E. coli gpt gene by the intact serp2 open reading frame. The Serp2(-) mutant virus replicated with wild-type kinetics both in rabbit fibroblasts and a rabbit CD4(+) T-cell line (RL5). Moderate reduction of cell surface levels of major histocompatibility complex I was observed after infection with wild-type or Serp2(-) mutant myxoma virus, and both produced white pocks on the chorioallantoic membrane of the chick embryo. After the infection of European rabbits, the Serp2(-) mutant virus proved to be highly attenuated compared to wild-type myxoma virus, as demonstrated by the clinical course of myxomatosis and the survival rates of infected animals. Pathohistological examinations revealed that infection with wild-type myxoma virus resulted in a blockade of the inflammatory response at the vascular level. In contrast, rapid inflammatory reactions occurred upon infection with the Serp2( ) mutant virus. Furthermore, lymphocytes in lymph nodes derived from animals inoculated with Serp2 mutant virus were shown to rapidly undergo apoptosis. We postulate that the virulence of myxoma virus in the European rabbit can be partially attributed to an impairment of host inflammatory processes and to the prevention of apoptosis in lymphocytes. The weakening of host defense is directly linked to serp2 gene function and is likely to involve the inhibition of IL-1beta converting-enzyme-dependent pathways. PMID- 9733820 TI - An envelope modification that renders a primary, neutralization-resistant clade B human immunodeficiency virus type 1 isolate highly susceptible to neutralization by sera from other clades. AB - SF162 is a primary (PR), non-syncytium-inducing, macrophagetropic human immunodeficiency virus type 1 (HIV-1) clade B isolate which is resistant to antibody-mediated neutralization. Deletion of the first or second hypervariable envelope gp120 region (V1 or V2 loop, respectively) of this virus does not abrogate its ability to replicate in peripheral blood mononuclear cells and primary macrophages, nor does it alter its coreceptor usage profile. The mutant virus with the V1 loop deletion, SF162DeltaV1, remains as resistant to antibody mediated neutralization as the wild-type virus SF162. In contrast, the mutant virus with the V2 loop deletion, SF162DeltaV2, exhibits enhanced susceptibility to neutralization by certain monoclonal antibodies whose epitopes are located within the CD4-binding site and conserved regions of gp120. More importantly, SF162DeltaV2 is now up to 170-fold more susceptible to neutralization than SF162 by sera collected from patients infected with clade B HIV-1 isolates. In addition, it becomes susceptible to neutralization by sera collected from patients infected with clade A, C, D, E, and F HIV-1 isolates. These findings suggest that the V2, but not the V1, loop of SF162 shields an as yet unidentified region of the HIV envelope rich in neutralization epitopes and that the overall structure of this region appears to be conserved among clade B, C, D, E, and F HIV-1 PR isolates. PMID- 9733821 TI - Env-independent protection induced by live, attenuated simian immunodeficiency virus vaccines. AB - Live attenuated simian immunodeficiency viruses (SIV), such as nef deletion mutants, are the most effective vaccines tested in the SIV-macaque model so far. To modulate the antiviral immune response induced by live attenuated SIV vaccines, we had previously infected rhesus monkeys with a nef deletion mutant of SIV expressing interleukin 2 (SIV-IL2) (B. R. Gundlach, H. Linhart, U. Dittmer, S. Sopper, S. Reiprich, D. Fuchs, B. Fleckenstein, G. Hunsmann, S. Stahl-Hennig, and K. Uberla, J. Virol. 71:2225-2232, 1997). In the present study, SIV-IL2 infected macaques and macaques infected with the nef deletion mutant SIVDeltaNU were challenged with pathogenic SIV 9 to 11 months postvaccination. In contrast to the results with naive control monkeys, no challenge virus could be isolated from the SIV-IL2- and SIVDeltaNU-infected macaques. However, challenge virus sequences could be detected by nested PCR in some of the vaccinated macaques. To determine the role of immune responses directed against Env of SIV, four vaccinated macaques were rechallenged with an SIV-murine leukemia virus (MLV) hybrid in which the env gene of SIV had been functionally replaced by the env gene of amphotropic MLV. All vaccinated macaques were protected from productive infection with the SIV-MLV hybrid in the absence of measurable neutralizing antibodies, while two naive control monkeys were readily infected. Since the SIV MLV hybrid uses the MLV Env receptor Pit2 and not CD4 and a coreceptor for virus entry, chemokine inhibition and receptor interference phenomena were not involved in protection. These results indicate that the protective responses induced by live attenuated SIV vaccines can be independent of host immune reactions directed against Env. PMID- 9733822 TI - Common themes of antibody maturation to simian immunodeficiency virus, simian human immunodeficiency virus, and human immunodeficiency virus type 1 infections. AB - Characterization of virus-specific immune responses to human immunodeficiency virus type 1 (HIV-1) and simian immunodeficiency virus (SIV) is important to understanding the early virus-host interactions that may determine the course of virus infection and disease. Using a comprehensive panel of serological assays, we have previously demonstrated a complex and lengthy maturation of virus specific antibody responses elicited by attenuated strains of SIV that was closely associated with the development of protective immunity. In the present study, we expand these analyses to address several questions regarding the nature of the virus-specific antibody responses to pathogenic SIV, SIV/HIV-1 (SHIV), and HIV-1 infections. The results demonstrate for the first time a common theme of antibody maturation to SIV, SHIV, and HIV-1 infections that is characterized by ongoing changes in antibody titer, conformational dependence, and antibody avidity during the first 6 to 10 months following virus infection. We demonstrate that this gradual evolution of virus-specific antibody responses is independent of the levels of virus replication and the pathogenicity of the infection viral strain. While the serological assays used in these studies were useful in discriminating between protective and nonprotective antibody responses during evaluation of vaccine efficacy with attenuated SIV, these same assays do not distinguish the clinical outcome of infection in pathogenic SIV, SHIV, or HIV-1 infections. These results likely reflect differences in the immune mechanisms involved in mediating protection from virus challenge compared to those that control an established viral infection, and they suggest that additional characteristics of both humoral and cellular responses evolve during this early immune maturation. PMID- 9733823 TI - Encapsidation of viral DNA requires the adenovirus L1 52/55-kilodalton protein. AB - Previous work demonstrated that the adenovirus L1 52/55-kDa protein is required for assembly of viral particles, although its exact role in the assembly process is unclear. The 52/55-kDa protein's early expression, however, suggests that it might have other roles at earlier times during infection. To uncover any role the 52/55-kDa protein might have at early times and to better characterize its role in assembly, a mutant adenovirus incapable of expressing the 52/55-kDa protein was constructed (H5pm8001). Analysis of the onset and extent of DNA replication and late protein synthesis revealed that H5pm8001-infected 293 cells entered the late stage of infection at the same time as did adenovirus type 5 (Ad5)-infected cells. Interestingly, H5pm8001-infected cells displayed slightly lower levels of replicated viral DNA and late proteins, suggesting that although not required, the 52/55-kDa protein does augment these activities during infection. Analysis of transcripts produced from the major late and IVa2 promoters indicated a slight reduction in H5pm8001-infected compared to Ad5-infected cells at 18 h postinfection that was not apparent at later times. Analysis of particles formed in H5pm8001 cells revealed that empty capsids could form, suggesting that the 52/55-kDa protein does not function as a scaffolding protein. Subsequent characterization of these particles demonstrated that they lacked any associated viral DNA. These findings indicate that the 52/55 kDa-protein is required to mediate stable association between the viral DNA and empty capsid and suggest that it functions in the DNA encapsidation process. PMID- 9733824 TI - Genetic variation in a human immunodeficiency virus type 2 live-virus Macaca nemestrina vaccine model. AB - Four pigtailed macaques were inoculated with an infectious, apathogenic human immunodeficiency virus type 2 (HIV-2) molecular clone (HIV-2KR) and subsequently challenged with a highly pathogenic strain, HIV-2287, together with two naive control animals. After challenge, two animals inoculated with a high dose of the immunizing strain were protected from CD4 decline and immunodeficiency. To examine the role of genetic heterogeneity in protection, fragments of the env gene were amplified from peripheral blood mononuclear cell DNA and plasma RNA of challenged animals by PCR, examined by using a heteroduplex tracking assay (HTA), and sequenced. By HTA, variation was detected principally within the V1 and V2 regions of envelope. Extent of variation in viral DNA clones as assessed by HTA correlated with inoculum size, as did the degree of variation in sequences of clones derived from viral DNA. Conversely, a rapid reduction in the number of plasma viral RNA variants was noted by HTA at 8 weeks postinfection in protected animals; this reduction was not present in naive or unprotected macaques. Sequences derived from plasma viral RNA were found to be more closely related than corresponding viral DNA sequences, and protection correlated with a significant reduction in variation in plasma RNA sequences in animals given the identical inocula of HIV-2287. Nonsynonymous mutations were significantly less prevalent in the protected animals. An additional potential glycosylation site was predicted to be present in the V2 region in all but one clone, and amino acid signatures related to protection were identified in viral DNA and RNA clones within both the V1 and V2 regions. Examination of the role of viral variation in this HIV-2 live-virus vaccine model may provide valuable insights into immunopathogenesis. PMID- 9733825 TI - Analysis of constructed E gene mutants of mouse hepatitis virus confirms a pivotal role for E protein in coronavirus assembly. AB - Expression studies have shown that the coronavirus small envelope protein E and the much more abundant membrane glycoprotein M are both necessary and sufficient for the assembly of virus-like particles in cells. As a step toward understanding the function of the mouse hepatitis virus (MHV) E protein, we carried out clustered charged-to-alanine mutagenesis on the E gene and incorporated the resulting mutations into the MHV genome by targeted recombination. Of the four possible clustered charged-to-alanine E gene mutants, one was apparently lethal and one had a wild-type phenotype. The two other mutants were partially temperature sensitive, forming small plaques at the nonpermissive temperature. Revertant analyses of these two mutants demonstrated that the created mutations were responsible for the temperature-sensitive phenotype of each and provided support for possible interactions among E protein monomers. Both temperature sensitive mutants were also found to be markedly thermolabile when grown at the permissive temperature, suggesting that there was a flaw in their assembly. Most significantly, when virions of one of the mutants were examined by electron microscopy, they were found to have strikingly aberrant morphology in comparison to the wild type: most mutant virions had pinched and elongated shapes that were rarely seen among wild-type virions. These results demonstrate an important, probably essential, role for the E protein in coronavirus morphogenesis. PMID- 9733826 TI - Mortality among human immunodeficiency virus type 2-positive villagers in rural Guinea-Bissau is correlated with viral genotype. AB - We present the results of a 6-year study of 131 human immunodeficiency virus (HIV) type 2 (HIV-2)-infected individuals from a rural population in Guinea Bissau. Proviral DNA sequences 1.3 kb in length were obtained from each individual and, together with clinical data, including proviral load and CD4 and CD8 levels, were used to assess whether viral genotype influences clinical outcome. With a phylogenetic model, a correlation was found between viral genotype and mortality; this correlation was not due to confounding factors, such as age-specific viral strains or cohabitation of patients. The data provide strong evidence for the involvement of viral genetic factors in determining HIV disease progression in vivo. The pattern of association found suggests that virulence factors are multiple and scattered throughout the HIV-2 genome and can be rapidly gained or lost by the virus through a combination of mutation and recombination. These findings may lead to the identification of viral determinants of HIV disease progression. PMID- 9733827 TI - Role of the TRAF binding site and NF-kappaB activation in Epstein-Barr virus latent membrane protein 1-induced cell gene expression. AB - In this study, we investigated the induction of cellular gene expression by the Epstein-Barr Virus (EBV) latent membrane protein 1 (LMP1). Previously, LMP1 was shown to induce the expression of ICAM-1, LFA-3, CD40, and EBI3 in EBV-negative Burkitt lymphoma (BL) cells and of the epidermal growth factor receptor (EGF-R) in epithelial cells. We now show that LMP1 expression also increased Fas and tumor necrosis factor receptor-associated factor 1 (TRAF1) in BL cells. LMP1 mediates NF-kappaB activation via two independent domains located in its C terminal cytoplasmic tail, a TRAF-interacting site that associates with TRAF1, 2, -3, and -5 through a PXQXT/S core motif and a TRADD-interacting site. In EBV transformed B cells or transiently transfected BL cells, significant amounts of TRAF1, -2, -3, and -5 are associated with LMP1. In epithelial cells, very little TRAF1 is expressed, and only TRAF2, -3, and -5, are significantly complexed with LMP1. The importance of TRAF binding to the PXQXT/S motif in LMP1-mediated gene induction was studied by using an LMP1 mutant that contains alanine point mutations in this motif and fails to associate with TRAFs. This mutant, LMP1(P204A/Q206A), induced 60% of wild-type LMP1 NF-kappaB activation and had approximately 60% of wild-type LMP1 effect on Fas, ICAM-1, CD40, and LFA-3 induction. In contrast, LMP1(P204A/Q206A) was substantially more impaired in TRAF1, EBI3, and EGF-R induction. Thus, TRAF binding to the PXQXT/S motif has a nonessential role in up-regulating Fas, ICAM-1, CD40, and LFA-3 expression and a critical role in up-regulating TRAF1, EBI3, and EGF-R expression. Further, D1 LMP1, an LMP1 mutant that does not aggregate failed to induce TRAF1, EBI3, Fas, ICAM-1, CD40, and LFA-3 expression confirming the essential role for aggregation in LMP1 signaling. Overexpression of a dominant form of IkappaBalpha blocked LMP1 mediated TRAF1, EBI3, Fas, ICAM-1, CD40, and LFA-3 up-regulation, indicating that NF-kappaB is an important component of LMP1-mediated gene induction from both the TRAF- and TRADD-interacting sites. PMID- 9733828 TI - The coxsackievirus-adenovirus receptor protein can function as a cellular attachment protein for adenovirus serotypes from subgroups A, C, D, E, and F. AB - Attachment of an adenovirus (Ad) to a cell is mediated by the capsid fiber protein. To date, only the cellular fiber receptor for subgroup C serotypes 2 and 5, the so-called coxsackievirus-adenovirus receptor (CAR) protein, has been identified and cloned. Previous data suggested that the fiber of the subgroup D serotype Ad9 also recognizes CAR, since Ad9 and Ad2 fiber knobs cross-blocked each other's cellular binding. Recombinant fiber knobs and 3H-labeled Ad virions from serotypes representing all six subgroups (A to F) were used to determine whether the knobs cross-blocked the binding of virions from different subgroups. With the exception of subgroup B, all subgroup representatives cross-competed, suggesting that they use CAR as a cellular fiber receptor as well. This result was confirmed by showing that CAR, produced in a soluble recombinant form (sCAR), bound to nitrocellulose-immobilized virions from the different subgroups except subgroup B. Similar results were found for blotted fiber knob proteins. The subgroup F virus Ad41 has both short and long fibers, but only the long fiber bound sCAR. The sCAR protein blocked the attachment of all virus serotypes that bound CAR. Moreover, CHO cells expressing human CAR, in contrast to untransformed CHO cells, all specifically bound the sCAR-binding serotypes. We conclude therefore that Ad serotypes from subgroups A, C, D, E, and F all use CAR as a cellular fiber receptor. PMID- 9733829 TI - Adeno-associated virus Rep78 protein interacts with protein kinase A and its homolog PRKX and inhibits CREB-dependent transcriptional activation. AB - Adeno-associated virus (AAV) is a human parvovirus of the genus Dependovirus. AAV replication is largely restricted to cells which are coinfected with a helper virus. In the absence of a helper virus, the AAV genome can integrate into a specific chromosomal site where it remains latent until reactivated by superinfection of the host cell with an appropriate helper virus. Replication functions of AAV have been mapped to the Rep68 and Rep78 gene products. Rep proteins demonstrate DNA binding, endonuclease, and helicase activities and are involved in regulation of transcription from both AAV and heterologous promoters. AAV has been associated with suppression of oncogenicity in a range of viral and nonviral tumors. In this study we sought to identify and study cellular protein targets of AAV Rep, in order to develop a better understanding of the various activities of Rep. We used the yeast two-hybrid system to identify HeLa cell proteins that interact with AAV type 2 Rep78. We isolated several strongly interacting clones which were subsequently identified as PRKX (previously named PKX1), a recently described homolog of the protein kinase A (PKA) catalytic subunit (PKAc). The interaction was confirmed in vitro by using pMal-Rep pull down assays. The region of Rep78 which interacts was mapped to a C-terminal zinc finger-like domain; Rep68, which lacks this domain, did not interact with PRKX. PRKX demonstrated autophosphorylation and kinase activity towards histone H1 and a PKA oligopeptide target. Autophosphorylation was inhibited by interaction with Rep78. In transfection assays, a PRKX expression vector was shown to be capable of activating CREB-dependent transcription. This activation was suppressed by Rep78 but not by Rep68. Since PRKX is a close homolog of PKAc, we investigated whether Rep78 could interact directly with PKAc. pMal-Rep78 was found to associate with purified PKAc and inhibited its kinase activity. Cotransfection experiments demonstrated that Rep78 could block the activation of CREB by a PKAc expression vector. These experiments suggest that AAV may perturb normal cyclic AMP response pathways in infected cells. PMID- 9733830 TI - Importance of the positive-strand RNA secondary structure of a murine coronavirus defective interfering RNA internal replication signal in positive-strand RNA synthesis. AB - The RNA elements that are required for replication of defective interfering (DI) RNA of the JHM strain of mouse hepatitis virus (MHV) consist of three discontinuous genomic regions: about 0.46 to 0.47 kb from both terminal sequences and an internal 58-nucleotide (nt)-long sequence (58-nt region) present at about 0.9 kb from the 5' end of the DI genome. The internal region is important for positive-strand DI RNA synthesis (Y. N. Kim and S. Makino, J. Virol. 69:4963 4971, 1995). We further characterized the 58-nt region in the present study and obtained the following results. (i) The positive-strand RNA structure in solution was comparable with that predicted by computer modeling. (ii) Positive-strand RNA secondary structure, but not negative-strand RNA structure, was important for the biological function of the region. (iii) The biological function had a sequence specific requirement. We discuss possible mechanisms by which the internal cis acting signal drives MHV positive-strand DI RNA synthesis. PMID- 9733831 TI - An orphan seven-transmembrane domain receptor expressed widely in the brain functions as a coreceptor for human immunodeficiency virus type 1 and simian immunodeficiency virus. AB - Both CD4 and an appropriate coreceptor are necessary for infection of cells by human immunodeficiency virus type 1 (HIV-1) and most strains of HIV-2. The chemokine receptors CCR5 and CXCR4 are the major HIV-1 coreceptors, although some virus strains can also utilize alternative coreceptors such as CCR3 to infect cells. In contrast, most if not all simian immunodeficiency virus (SIV) strains use CCR5 as a coreceptor, and many SIV strains can use CCR5 independently of CD4. In addition, several orphan seven-transmembrane receptors which can serve as HIV 1 and SIV coreceptors have been identified. Here we report that APJ, an orphan seven-transmembrane domain receptor with homology to the angiotensin receptor family, functions as a coreceptor for a number of HIV-1 and SIV strains. APJ was expressed widely in the human brain and in NT2N neurons. APJ transcripts were also detected by reverse transcription-PCR in the CD4-positive T-cell line C8166, but not in peripheral blood leukocytes, microglia, phytohemagglutinin (PHA)- or PHA/interleukin-2-stimulated peripheral blood mononuclear cells, monocytes, or monocyte-derived macrophages. The widespread distribution of APJ in the central nervous system coupled with its use as a coreceptor by some HIV-1 strains indicates that it may play a role in neuropathogenesis. PMID- 9733832 TI - Deoxyribonucleoside triphosphate pool imbalances in vivo are associated with an increased retroviral mutation rate. AB - Deoxyribonucleoside triphosphate (dNTP) pool imbalances are associated with an increase in the rate of misincorporation and hypermutation during in vitro reverse transcription reactions. However, the effects of in vivo dNTP pool imbalances on the accuracy of reverse transcription are unknown. We sought to determine the effects of in vivo dNTP pool imbalances on retroviral mutation rates and to test our hypothesis that 3'-azido-3'-deoxythymidine (AZT) increases the retroviral mutation rates through induction of dNTP pool imbalances. D17 cells were treated with thymidine, hydroxyurea (HU), or AZT, and the effects on in vivo dNTP pools were measured. Thymidine and HU treatments induced significant dNTP pool imbalances. In contrast, AZT treatment had very little effect on the dNTP pools. The effects of in vivo dNTP pool imbalances induced by thymidine and HU treatments on the retroviral mutation rates were also determined. Spleen necrosis virus (SNV)-based and murine leukemia virus (MLV)-based retroviral vectors that expressed the lacZ mutant reporter gene were used. The frequencies of inactivating mutations introduced in the lacZ gene in a single replication cycle provided a measure of the retroviral mutation rates. Treatment of D17 target cells with 500 microM thymidine increased the SNV and MLV mutant frequencies 4.7- and 4-fold, respectively. Treatment of D17 target cells with 2 mM HU increased the SNV and MLV mutant frequencies 2.1- and 2.7-fold, respectively. These results demonstrate that dNTP pool imbalances are associated with an increase in the in vivo retroviral mutation rates, but AZT treatment results in an increase in the retroviral mutation rates by a mechanism not involving alterations in dNTP pools. PMID- 9733833 TI - Analysis of minimal human immunodeficiency virus type 1 gag coding sequences capable of virus-like particle assembly and release. AB - We have constructed a series of human immunodeficiency virus (HIV) gag mutants by progressive truncation of the gag coding sequence from the C terminus and have combined these mutants with an assembly-competent matrix domain deletion mutation (DeltaMA). By using several methods, the particle-producing capabilities of each mutant were examined. Our analysis indicated that truncated Gag precursors lacking most of C-terminal gag gene products assembled and were released from 293T cells. Additionally, a mutant with a combined deletion of the MA (DeltaMA) and p6 domains even produced particles at levels comparable to that of the wild type (wt) virus. However, most mutants derived from combination of the DeltaMA and the C-terminal truncation mutations did not release particles as well as the wt. Our smallest HIV gag gene product capable of virus-like particle formation was a 28-kDa protein which consists of a few MA amino acids and the CA-p2 domain. Sucrose density gradient fractionation analysis indicated that most mutants exhibited a wt retrovirus particle density. Exceptions to this rule were mutants with an intact MA domain but deleted downstream of the p2 domains. These C terminal truncation mutants possessed particle densities of 1.13 to 1.15 g/ml, lower than that of the wt. The N-terminal portions of the CA domain, which have been shown to be dispensable for core assembly, became critical when most of the MA domain was deleted, suggesting a requirement for an intact CA domain to assemble and release particles. PMID- 9733835 TI - trans-encapsidation of a poliovirus replicon by different picornavirus capsid proteins. AB - A trans-encapsidation assay was established to study the specificity of picornavirus RNA encapsidation. A poliovirus replicon with the luciferase gene replacing the capsid protein-coding region was coexpressed in transfected HeLa cells with capsid proteins from homologous or heterologous virus. Successful trans-encapsidation resulted in assembly and production of virions whose replication, upon subsequent infection of HeLa cells, was accompanied by expression of luciferase activity. The amount of luciferase activity was proportional to the amount of trans-encapsidated virus produced from the cotransfection. When poliovirus capsid proteins were supplied in trans, >2 x 10(6) infectious particles/ml were produced. When coxsackievirus B3, human rhinovirus 14, mengovirus, or hepatitis A virus (HAV) capsid proteins were supplied in trans, all but HAV showed some encapsidation of the replicon. The overall encapsidation efficiency of the replicon RNA by heterologous capsid proteins was significantly lower than when poliovirus capsid was used. trans encapsidated particles could be completely neutralized with specific antisera against each of the donor virus capsids. The results indicate that encapsidation is regulated by specific viral nucleic acid and protein sequences. PMID- 9733834 TI - E1B 55-kilodalton-associated protein: a cellular protein with RNA-binding activity implicated in nucleocytoplasmic transport of adenovirus and cellular mRNAs. AB - The adenovirus type 5 (Ad5) early 1B 55-kDa protein (E1B-55kDa) is a multifunctional phosphoprotein that regulates viral DNA replication and nucleocytoplasmic RNA transport in lytically infected cells. In addition, E1B 55kDa provides functions required for complete oncogenic transformation of rodent cells in cooperation with the E1A proteins. Using the far-Western technique, we have isolated human genes encoding E1B-55kDa-associated proteins (E1B-APs). The E1B-AP5 gene encodes a novel nuclear RNA-binding protein of the heterogeneous nuclear ribonucleoprotein (hnRNP) family that is highly related to hnRNP-U/SAF-A. Immunoprecipitation experiments indicate that two distinct segments in the 55-kDa polypeptide which partly overlap regions responsible for p53 binding are required for complex formation with E1B-AP5 in Ad-infected cells and that this protein interaction is modulated by the adenovirus E4orf6 protein. Expression of E1B-AP5 efficiently interferes with Ad5 E1A/E1B-mediated transformation of primary rat cells. Furthermore, stable expression of E1B-AP5 in Ad-infected cells overcomes the E1B-dependent inhibition of cytoplasmic host mRNA accumulation. These data suggest that E1B-AP5 might play a role in RNA transport and that this function is modulated by E1B-55kDa in Ad-infected cells. PMID- 9733836 TI - The Epstein-Barr virus Rta protein activates lytic cycle genes and can disrupt latency in B lymphocytes. AB - The transition of Epstein-Barr virus (EBV) from latency into the lytic cycle is associated with the expression of two immediate-early viral genes, BZLF1 and BRLF1. Overexpression of ZEBRA, the product of BZLF1, is sufficient to disrupt latency in B lymphocytes and epithelial cells by stimulating expression of lytic cycle genes, including BRLF1. The BRLF1 product Rta functions as a transcriptional activator in both B lymphocytes and epithelial cells. However, Rta has recently been reported to disrupt latency in an epithelial specific manner (S. Zalani, E. Holley-Guthrie, and S. Kenney, Proc. Natl. Acad. Sci. USA 93:9194-9199, 1996). Here we demonstrate that expression of Rta is also sufficient for disruption of latency in a permissive B-cell line. In HH514-16 cells, transfection of Rta leads to synthesis of ZEBRA, viral DNA replication, and late gene expression. However, Rta by itself is less potent than ZEBRA in the ability to activate most early and late lytic cycle genes. In light of previous work implicating ZEBRA in the activation of Rta, we suggest a cooperative model for EBV entry into the lytic cycle. Expression of either BZLF1 or BRLF1 triggers expression of the other immediate-early factor, and together these activators act individually or in synergy on downstream targets to activate the viral lytic cycle. PMID- 9733837 TI - A virus-encoded RNA polymerase purified from baculovirus-infected cells. AB - A DNA-dependent RNA polymerase was purified to homogeneity, starting from insect cells infected with the baculovirus Autographa californica nuclear polyhedrosis virus (AcNPV). The purified polymerase supported accurate and specific transcription from late and very late promoters but was not active on viral early promoters. Thus, promoter recognition is an integral function of the purified enzyme. The purified RNA polymerase was composed of only four equimolar subunits, which makes it the simplest DNA-directed RNA polymerase from a eukaryotic source described so far. Amino-terminal protein sequencing, peptide fingerprinting, and immunochemical analyses were used to identify the four subunits, all of which are virus encoded. Overexpression of the four viral proteins (LEF-8, LEF-4, LEF-9, and p47) in baculovirus-infected cells resulted in a significant increase in the levels of RNA polymerase produced in the infected cells. Thus, the overexpression data are consistent with our identification of the RNA polymerase subunits. PMID- 9733838 TI - Inactivation of human immunodeficiency virus type 1 infectivity with preservation of conformational and functional integrity of virion surface proteins. AB - Whole inactivated viral particles have been successfully used as vaccines for some viruses, but procedures historically used for inactivation can denature virion proteins. Results have been inconsistent, with enhancement of disease rather than protection seen in some notable instances following vaccination. We used the compound 2,2'-dithiodipyridine (aldrithiol-2; AT-2) to covalently modify the essential zinc fingers in the nucleocapsid (NC) protein of human immunodeficiency virus type 1 (HIV-1) or simian immunodeficiency virus (SIV) virions, thereby inactivating infectivity. The inactivated virus was not detectably infectious in vitro (up to 5 log units of inactivation). However, in contrast to virions inactivated by conventional methods such as heat or formalin treatment, viral and host cell-derived proteins on virion surfaces retained conformational and functional integrity. Thus, immunoprecipitation of AT-2 treated virions was comparable to precipitation of matched untreated virus, even when using antibodies to conformational determinants on gp120. AT-2 inactivated virions bound to CD4(+) target cells and mediated virus-induced, CD4-dependent "fusion from without" comparably to native virions. However, viral entry assays demonstrated that the viral life cycle of AT-2-treated virions was arrested before initiation of reverse transcription. The major histocompatibility complex (MHC) class II molecules on the surface of AT-2-treated virions produced from MHC class II-expressing cells retained the ability to support class II-dependent, superantigen-triggered proliferative responses by resting T lymphocytes. These findings indicate that inactivation via this method results in elimination of infectivity with preservation of conformational and functional integrity of virion surface proteins, including both virally encoded determinants and proteins derived from the host cells in which the virus was produced. Such inactivated virions should provide a promising candidate vaccine antigen and a useful reagent for experimentally probing the postulated involvement of virion surface proteins in indirect mechanisms of HIV-1 pathogenesis. PMID- 9733839 TI - Biochemical activities of minute virus of mice nonstructural protein NS1 are modulated In vitro by the phosphorylation state of the polypeptide. AB - NS1, the 83-kDa major nonstructural protein of minute virus of mice (MVM), is a multifunctional nuclear phosphoprotein which is required in a variety of steps during progeny virus production, early as well as late during infection. NS1 is the initiator protein for viral DNA replication. It binds specifically to target DNA motifs; has site-specific single-strand nickase, intrinsic ATPase, and helicase activities; trans regulates viral and cellular promoters; and exerts cytotoxic stress on the host cell. To investigate whether these multiple activities of NS1 depend on posttranslational modifications, in particular phosphorylation, we expressed His-tagged NS1 in HeLa cells by using recombinant vaccinia viruses, dephosphorylated it at serine and threonine residues with calf intestine alkaline phosphatase, and compared the biochemical activities of the purified un(der)phosphorylated (NS1(O)) and the native (NS1(P)) polypeptides. Biochemical analyses of replicative functions of NS1(O) revealed a severe reduction of intrinsic helicase activity and, to a minor extent, of ATPase and nickase activities, whereas its affinity for the target DNA sequence [ACCA]2-3 was enhanced compared to that of NS1(P). In the presence of endogenous protein kinases found in replication extracts, NS1(O) showed all functions necessary for resolution and replication of the 3' dimer bridge, indicating reactivation of NS1(O) by rephosphorylation. Partial reactivation of the helicase activity was found as well when NS1(O) was incubated with protein kinase C. PMID- 9733840 TI - Processing of proteinase precursors and their effect on hepatitis A virus particle formation. AB - Proteolytic processing of the picornaviral polyprotein mediated by the differential action of virus-encoded proteinase(s) is pivotal to both RNA genome replication and capsid formation. Possibly to enlarge the array of viral proteins, picornaviral polyprotein processing results in intermediate and mature products which apparently have distinct functions within the viral life cycle. For hepatitis A virus (HAV), we report here on the autoproteolysis of precursor polypeptides comprising the only viral proteinase, 3Cpro, and on their role in viral particle formation. Following transient expression of a nested set of 3Cpro containing proteins (P3, 3ABC, 3BCD, 3CD, 3BC, and 3C) in eukaryotic cells, the extent of processing was determined by analyzing the cleavage products. The 3C/3D site was more efficiently cleaved than those at the 3A/3B and 3B/3C sites, leading to the accumulation of the intermediate product 3ABC. In the absence of 3A from the precursor, cleavage at the 3B/3C site was further reduced and a switch to an alternative 3C/3D site was observed. Coexpression of various parts of P3 with the precursor of the viral structural proteins P1-2A showed that all 3C-containing intermediates cleaved P1-2A with almost equal efficiency; however, viral particles carrying the neutralizing epitope form much more readily in the presence of the complete P3 domain than with parts of it. These data support the notion that efficient liberation of structural proteins from P1-2A is necessary but not sufficient for productive HAV capsid formation and suggest that the polypeptides flanking 3Cpro promote the assembly of viral particles. PMID- 9733841 TI - Phylogenetic analysis of the entire genome of influenza A (H3N2) viruses from Japan: evidence for genetic reassortment of the six internal genes. AB - Nucleotide sequences of all eight RNA segments of 10 human H3N2 influenza viruses isolated during a 5-year period from 1993 to 1997 were determined and analyzed phylogenetically in order to define the evolutionary pathways of all genes in a parallel fashion. It was evident that the hemagglutinin and neuraminidase genes of these viruses evolved essentially in a single lineage and that amino acid changes accumulated sequentially with respect to time. In contrast, amino acid differences in the internal proteins were erratic and did not accumulate over time. Parallel analysis of the phylogenetic patterns of all genes revealed that the evolutionary pathways of the six internal genes were not linked to the surface glycoproteins. Genes coding for the basic polymerase-1, nucleoprotein, and matrix proteins of 1997 isolates were closest phylogenetically to those of earlier isolates of 1993 and 1994. Furthermore, all six internal genes of four viruses isolated in the 1995 epidemic season consistently divided into two distinct branch clusters, and two 1995 isolates contained PB2 genes apparently originating from those of viruses before 1993. It was apparent that the lack of correlation between the topologies of the phylogenetic trees of the genes coding for the surface glycoproteins and internal proteins was a reflection of genetic reassortment among human H3N2 viruses. This is the first evidence demonstrating the occurrence of genetic reassortment involving the internal genes of human H3N2 viruses. Furthermore, internal protein variability coincided with marked increases in the activity of H3N2 viruses in 1995 and 1997. PMID- 9733842 TI - Establishment and characterization of Japanese encephalitis virus-specific, human CD4(+) T-cell clones: flavivirus cross-reactivity, protein recognition, and cytotoxic activity. AB - We analyzed the CD4(+) T-lymphocyte responses of two donors who had received Japanese encephalitis virus (JEV) vaccine 6 or 12 months earlier. Bulk culture proliferation assays showed that peripheral blood mononuclear cells (PBMC) responded to JEV antigens (Ag) but also responded at lower levels to West Nile virus (WNV) and dengue virus type 1, 2, and 4 (D1V, D2V, and D4V, respectively) Ag. Five JEV-specific CD4(+) human T-cell clones and one subclone were established from PBMC of these two donors. Two clones responded to WNV Ag as well as to JEV Ag, whereas the others responded only to JEV Ag. Three of five CD4(+) T cell clones had JEV-specific cytotoxic activity and recognized E protein. The HLA restriction of the JEV-specific T-cell clones was examined. Three clones were HLA DR4 restricted, one was HLA-DQ3 restricted, and the HLA restriction of one clone was not determined. T-cell receptor analysis showed that these clones expressed different T-cell receptors, suggesting that they originated from different T lymphocytes. These results indicate that JEV vaccine induces JEV-specific and flavivirus-cross-reactive CD4(+) T lymphocytes and that these T lymphocytes recognize E protein. The functions and HLA restriction patterns of these T lymphocytes are, however, heterogeneous. PMID- 9733843 TI - Comparison of the neurovirulence of a vaccine and a wild-type mumps virus strain in the developing rat brain. AB - Prior to the adoption of widespread vaccination programs, mumps virus was the leading cause of virus-induced central nervous system (CNS) disease. Mumps virus associated CNS complications in vaccinees continue to be reported; outside the United States, some of these complications have been attributed to vaccination with insufficiently attenuated neurovirulent vaccine strains. The development of potentially neurovirulent, live, attenuated mumps virus vaccines stems largely from the lack of an animal model that can reliably predict the neurovirulence of mumps virus vaccine candidates in humans. The lack of an effective safety test with which to measure mumps virus neurovirulence has also hindered analysis of the neuropathogenesis of mumps virus infection and the identification of molecular determinants of neurovirulence. In this report we show, for the first time, that mumps virus infection of the neonatal rat leads to developmental abnormalities in the cerebellum due to cerebellar granule cell migration defects. The incidence of the cerebellar abnormalities and other neuropathological and clinical outcomes of mumps virus infection of the neonatal rat brain demonstrated the ability of this model to distinguish neurovirulent (Kilham) from nonneurovirulent (Jeryl Lynn) mumps virus strains. Thus, this neonatal rat model may prove useful in evaluating the neurovirulence potential of new live, attenuated vaccine strains and may also be of value in elucidating the molecular basis of mumps virus neurovirulence. PMID- 9733844 TI - The Epstein-Barr virus immediate-early gene product, BRLF1, interacts with the retinoblastoma protein during the viral lytic cycle. AB - Retinoblastoma protein (Rb) is a key regulator of cellular proliferation, controlling entry into G1/S in the cell cycle, largely through its action in binding the cellular transcription factor E2F, which activates genes important in DNA synthesis. Small DNA tumor viruses encode gene products which can functionally inactivate Rb, promoting cellular proliferation and viral DNA synthesis. In this study, the Epstein-Barr virus (EBV) immediate-early lytic gene product, BRLF1 (R), is shown to bind Rb in vivo, shortly after induction of the viral lytic cycle in EBV-infected Akata cells. Furthermore, the temporal kinetics of R-Rb interaction correlate with displacement of E2F1 from Rb. Mapping of the domains required for the interaction of R and Rb proteins reveals that R binds specifically to the N terminus of Rb, outside the Rb pocket, and that the first 200 amino acids of R are required for this interaction. The interaction of R and Rb may initiate cell cycle progression and facilitate viral DNA synthesis during lytic replication. PMID- 9733845 TI - Protection against lethal encephalomyocarditis virus infection in the absence of serum-neutralizing antibodies. AB - Although the ability of serum-neutralizing antibodies to protect against picornavirus infection is well established, the contribution of cell-mediated immunity to protection is uncertain. Using major histocompatibility complex class II-deficient (RHAbeta-/-) mice, which are unable to mediate CD4(+) T-lymphocyte dependent humoral responses, we demonstrated antibody-independent protection against lethal encephalomyocarditis virus (EMCV) infection in the natural host. The majority of RHAbeta-/- mice inoculated with 10(4) PFU of attenuated Mengo virus (vMC24) resolved infection and were resistant to lethal challenge with the highly virulent, serotypically identical cardiovirus, EMCV. Protection in these mice was in the absence of detectable serum-neutralizing antibodies. Depletion of CD8(+) T lymphocytes prior to lethal EMCV challenge ablated protection in vMC24 immunized RHAbeta-/- mice. The CD8(+) T-lymphocyte-dependent protection observed in vivo may, in part, be the result of cytotoxic T-lymphocyte (CTL) activity, as CD8(+) T splenocytes exhibited in vitro cytolysis of EMCV-infected targets. The existence of virus-specific CD8(+) T-lymphocyte memory in these mice was demonstrated by increased expression of cell surface activation markers CD25, CD69, CD71, and CTLA-4 following antigen-specific reactivation in vitro. Although recall response in vMC24-immunized RHAbeta-/- mice was intact and effectual shortly after immunization, protection abated over time, as only 3 of 10 vMC24 immunized RHAbeta-/- mice survived when rechallenged 90 days later. The present study demonstrating CD8(+) T-lymphocyte-dependent protection in the absence of serum-neutralizing antibodies, coupled with our previous results indicating that vMC24-specific CD4(+) T lymphocytes confer protection against lethal EMCV in the absence of prophylactic antibodies, suggests the existence of nonhumoral protective mechanisms against picornavirus infections. PMID- 9733847 TI - In vivo selection of Rous sarcoma virus mutants with randomized sequences in the packaging signal. AB - Retrovirus genomes contain a sequence at the 5' end which directs their packaging into virions. In Rous sarcoma virus, previous studies have identified important segments of the packaging signal, Psi, and support elements of a secondary structure prediction. To further characterize this sequence, we used an in vivo selection strategy to test large collections of mutants. We generated pools of full-length viral DNA molecules with short stretches of random sequence in Psi and transfected each pool into avian cells. Resulting infectious virus was allowed to spread by multiple passages, so that sequences could compete and the best could be selected. This method provides information on the kinds of sequences allowed, as well as those that are most fit. Several predicted stem loop structures in Psi were tested. A stem at the base of element O3 was highly favored; only sequences which maintained base pairing were selected. Two other stems, at the base and in the middle of element L3, were not conserved: neither base pairing nor sequence was maintained. A single mutation, G213U, was seen upstream of the randomized region in all selected L3 stem mutants; we interpret this to mean that it compensates for the defects in L3. Randomized mutations adjacent to G213 maintained the wild-type base composition but not its sequence. The kissing-loop sequence at end of L3, postulated to function in genome dimerization, was not required for infectivity but was selected for over time. Finally, a deletion of L3 was constructed and found to be poorly infectious. PMID- 9733846 TI - Molecular and cellular analysis of human immunodeficiency virus-induced apoptosis in lymphoblastoid T-cell-line-expressing wild-type and mutated CD4 receptors. AB - We have previously shown that the presence of the CD4 cytoplasmic tail is critical for human immunodeficiency virus (HIV)-induced apoptosis (J. Corbeil, M. Tremblay, and D. D. Richman, J. Exp. Med. 183:39-48, 1996). We have pursued our investigation of the role of the CD4 transduction pathway in HIV-induced apoptosis. To do this, wild-type and mutant forms of the CD4 cytoplasmic tail were stably expressed in the lymphoblastoid T-cell line A2.01. Apoptosis was prevented when CD4 truncated at residue 402 was expressed; however, cells expressing mutated receptors that do not associate with p56(lck) (mutated at the dicysteine motif and truncated at residue 418) but which conserved proximal domains of the cytoplasmic tail underwent apoptosis like wild-type CD4. The differences between wild-type and mutated receptors in the induction of apoptosis were not related to levels of p56(lck) or NF-kappaB activation. Initial signaling through the CD4 receptor played a major role in the sensitization of HIV-infected T cells to undergo apoptosis. Incubation of HIV-infected cells with monoclonal antibody (MAb) 13B8-2, which binds to CD4 in a region critical for dimerization of the receptor, prevented apoptosis without inhibiting HIV replication. Moreover, the apoptotic process was not related to Fas-Fas ligand interaction; however, an antagonistic anti-Fas MAb (ZB-4) enhanced apoptosis in HIV-infected cells without inducing apoptosis in uninfected cells. These observations demonstrate that CD4 signaling mediates HIV-induced apoptosis by a mechanism independent of Fas-Fas ligand interaction, does not require p56(lck) signaling, and may involve a critical region for CD4 dimerization. PMID- 9733848 TI - Varicella-zoster virus (VZV) ORF32 encodes a phosphoprotein that is posttranslationally modified by the VZV ORF47 protein kinase. AB - Varicella-zoster virus (VZV) encodes five gene products that do not have homologs in herpes simplex virus. One of these genes, VZV open reading frame 32 (ORF32), is predicted to encode a protein of 16 kDa. VZV ORF32 protein was shown to be phosphorylated and located in the cytosol of virus-infected cells. Antibody to ORF32 protein immunoprecipitated 16- and 18-kDa phosphoproteins from VZV-infected cells. Since VZV encodes two protein kinases that might phosphorylate ORF32 protein, immunoprecipitations were performed with cells infected with VZV mutants unable to express either of the viral protein kinases. Cells infected with VZV unable to express the ORF66 protein kinase contained both the 16- and 18-kDa ORF32 phosphoproteins; however, cells infected with the VZV ORF47 protein kinase mutant showed only the 16-kDa ORF32 phosphoprotein. Treatment of [35S]methionine labeled proteins with calf intestine alkaline phosphatase resulted in a decrease in size of the ORF32 proteins from 16 and 18 kDa to 15 and 17 kDa, respectively. VZV unable to express ORF32 protein replicated in human melanoma cells to titers similar to those seen with parental virus; however, VZV unable to express ORF32 was impaired for replication in U20S osteosarcoma cells. Thus, VZV ORF32 protein is posttranslationally modified by the ORF47 protein kinase. Since the VZV ORF47 protein kinase has recently been shown to be critical for replication in human fetal skin and lymphocytes, its ability to modify the ORF32 protein suggests that the latter protein may have a role for VZV replication in human tissues. PMID- 9733850 TI - Carboxypeptidase D (gp180), a Golgi-resident protein, functions in the attachment and entry of avian hepatitis B viruses. AB - Carboxypeptidase D (gp180), one of many candidate receptors proposed for hepatitis B viruses (HBVs), was examined and found to be the actual cellular receptor for avian HBVs. This conclusion was based on the following observations: (i) gp180 was the only host protein that bound with high affinity to the pre-S ectodomain of the large duck hepatitis B virus (DHBV) envelope protein, which is known to be essential for virus infection; (ii) a pre-S subdomain which determines physical binding to gp180 was found to coincide with a domain functionally defined in infection competition experiments as a receptor binding domain; (iii) soluble gp180, lacking the membrane anchor, efficiently inhibited DHBV infection; (iv) efficient interspecies gp180-pre-S interaction was limited to the natural hosts of avian hepadnaviruses; and (v) expression of gp180 in a heterologous hepatoma cell line mediated cellular attachment and subsequent internalization of fluorescently labeled viral particles into vesicular structures. However, gp180 expression did not render transfected heterologous cells permissive for productive infection, suggesting that a species-specific coreceptor is required for fusion to complete viral entry. In contrast to the case for known virus receptors, gp180 was not detected on the hepatocyte cell surface but was found to be concentrated in the Golgi apparatus, from where it functions by cycling to and from the plasma membrane. PMID- 9733849 TI - Avian hepatitis B virus infection is initiated by the interaction of a distinct pre-S subdomain with the cellular receptor gp180. AB - Functionally relevant hepadnavirus-cell surface interactions were investigated with the duck hepatitis B virus (DHBV) animal model by using an in vitro infection competition assay. Recombinant DHBV pre-S polypeptides, produced in Escherichia coli, were shown to inhibit DHBV infection in a dose-dependent manner, indicating that monomeric pre-S chains were capable of interfering with virus-receptor interaction. Particle-associated pre-S was, however, 30-fold more active, suggesting that cooperative interactions enhance particle binding. An 85 amino-acid pre-S sequence, spanning about half of the DHBV pre-S chain, was characterized by deletion analysis as essential for maximal inhibition. Pre-S polypeptides from heron hepatitis B virus (HHBV) competed DHBV infection equally well despite a 50% difference in amino acid sequence and a much-reduced infectivity of HHBV for duck hepatocytes. These observations are taken to indicate (i) that the functionality of the DHBV pre-S subdomain, which interacts with the cellular receptor, is determined predominantly by a defined three dimensional structure rather than by primary sequence elements; (ii) that cellular uptake of hepadnaviruses is a multistep process involving more than a single cellular receptor component; and (iii) that gp180, a cellular receptor candidate unable to discriminate between DHBV and HHBV, is a common component of the cellular receptor complex for avian hepadnaviruses. PMID- 9733851 TI - Activation of the Epstein-Barr virus transcription factor BZLF1 by 12-O tetradecanoylphorbol-13-acetate-induced phosphorylation. AB - BZLF1 is a member of the extended AP-1 family of transcription factors which binds to specific BZLF1 sequence motifs within early Epstein-Barr virus (EBV) promoters and to closely related AP-1 motifs. BZLF1's activity is regulated at the transcriptional level as well as through protein interactions and posttranslational modifications. Phorbol esters or immunoglobulin cross-linking both reactivate EBV from latently infected B cells via transactivation of BZLF1. We report here that the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) is capable of inducing BZLF1's activity even further. The induction occurs at the posttranscriptional level and depends on a single serine residue located in the DNA binding domain of BZLF1. This serine residue (S186) is phosphorylated by protein kinase C in vitro and in vivo after stimulation with TPA. Phosphorylation of S186 per se interferes with the DNA binding affinity of BZLF1 in vitro but is mandatory for TPA-induced increase in DNA binding of BZLF1, as shown in gel retardation assays and reconstruction experiments with cellular extracts. In transcriptional reporter assays, S186 is essential for the activation of BZLF1 by TPA. Presumably, a yet-to-be-identified cellular factor restores the DNA binding affinity and enhances the transcriptional activity of S186-phosphorylated BZLF1, which is required to induce the lytic phase of EBV's life cycle. PMID- 9733852 TI - Transactivation by the E2 protein of oncogenic human papillomavirus type 31 is not essential for early and late viral functions. AB - The activation of transcription and of DNA replication are, in some cases, mediated by the same proteins. A prime example is the E2 protein of human papillomaviruses (HPVs), which binds ACCN6GGT sequences and activates heterologous promoters from multimerized binding sites. The E2 protein also has functions in replication, where it complexes with the virally encoded origin recognition protein, E1. Much of the information on these activities is based on transient-transfection assays as well as biochemical analyses; however, their importance in the productive life cycle of oncogenic HPVs remains unclear. To determine the contributions of these E2 functions to the HPV life cycle, a genetic analysis was performed by using an organotypic tissue culture model. HPV type 31 (HPV31) genomes that contained mutations in the N terminus of E2 (amino acid 73) were constructed; these mutants retained replication activities but were transactivation defective. Following transfection of normal human keratinocytes, these mutant genomes were established as stable episomes and expressed early viral transcripts at levels similar to those of wild-type HPV31. Upon differentiation in organotypic raft cultures, the induction of late gene expression and amplification of viral DNA were detected in cell lines harboring mutant genomes. Interestingly, only a modest reduction in late gene expression was observed in the mutant lines. We conclude that the transactivation function of E2 is not essential for the viral life cycle of oncogenic HPVs, although it may act to moderately augment late expression. Our studies suggest that the primary positive role of E2 in the viral life cycle is as a replication factor. PMID- 9733853 TI - Expression of measles virus V protein is associated with pathogenicity and control of viral RNA synthesis. AB - Nonstructural proteins encoded by measles virus (MV) include the V protein which is translated from an edited P mRNA. V protein is not associated with intracellular or released viral particles and has recently been found to be dispensable for MV propagation in cell culture (H. Schneider, K. Kaelin, and M. A. Billeter, Virology 227:314-322, 1997). Using recombinant MVs (strain Edmonston [ED]) genetically engineered to overexpress V protein (ED-V+) or to be deficient for V protein (ED-V-), we found that in the absence of V both MV-specific proteins and RNAs accumulated to levels higher than those in the parental MV molecular clone (ED-tag), whereas MV-specific gene expression was strongly attenuated in human U-87 glioblastomas cells after infection with ED-V+. The titers of virus released from these cells 48 h after infection with either V mutant virus were lower than those from cells infected with ED-tag. Similarly, significantly reduced titers of infectious virus were reisolated from lung tissue of cotton rats (Sigmodon hispidus) after intranasal infection with both editing mutants compared to titers isolated from ED-tag-infected animals. In cell culture, expression of V protein led to a redistribution of MV N protein in doubly transfected Cos-7 cells, indicating that these proteins form heterologous complexes. This interaction was further confirmed by using a two-hybrid approach with both proteins expressed as Gal4 or VP16 fusion products. Moreover, V protein efficiently competed complexes formed between MV N and P proteins. These findings indicate that V protein acts to balance accumulation of viral gene products in cell culture, and this may be dependent on its interaction with MV N protein. Furthermore, expression of V protein may contribute to viral pathogenicity in vivo. PMID- 9733854 TI - A protein encoded by the latency-related gene of bovine herpesvirus 1 is expressed in trigeminal ganglionic neurons of latently infected cattle and interacts with cyclin-dependent kinase 2 during productive infection. AB - Despite productive viral gene expression in the peripheral nervous system during acute infection, the bovine herpesvirus 1 (BHV-1) infection cycle is blocked in sensory ganglionic neurons and consequently latency is established. The only abundant viral transcript expressed during latency is the latency-related (LR) RNA. LR gene products inhibit S-phase entry, and binding of the LR protein (LRP) to cyclin A was hypothesized to block cell cycle progression. This study demonstrates LRP is a nuclear protein which is expressed in neurons of latently infected cattle. Affinity chromatography indicated that LRP interacts with cyclin dependent kinase 2 (cdk2)-cyclin complexes or cdc2-cyclin complexes in transfected human cells or infected bovine cells. After partial purification using three different columns (DEAE-Sepharose, Econo S, and heparin-agarose), LRP was primarily associated with cdk2-cyclin E complexes, an enzyme which is necessary for G1-to-S-phase cell cycle progression. During acute infection of trigeminal ganglia or following dexamethasone-induced reactivation, BHV-1 induces expression of cyclin A in neurons (L. M. Schang, A. Hossain, and C. Jones, J. Virol. 70:3807-3814, 1996). Expression of S-phase regulatory proteins (cyclin A, for example) leads to neuronal apoptosis. Consequently, we hypothesize that interactions between LRP and cell cycle regulatory proteins promote survival of postmitotic neurons during acute infection and/or reactivation. PMID- 9733855 TI - Persistent infection of Epstein-Barr virus-positive B lymphocytes by human herpesvirus 8. AB - In patients with Kaposi's sarcoma (KS), human herpesvirus 8 (HHV-8) can invariably be detected in KS tumor tissue and, at a lower frequency, in prostate tissue and peripheral blood B lymphocytes. Whereas the majority of KS spindle cells are latently infected by HHV-8, linear HHV-8 genomes characteristic for lytic infection are found predominantly in the peripheral blood cells of KS patients. In this study, we show that HHV-8 can stably infect B lymphocytes in vitro in the presence of Epstein-Barr virus (EBV). We were able to generate immortalized HHV-8(+)/EBV+ lymphoblastoid cell lines (LCLs) derived from peripheral blood mononuclear cells (PBMC) of EBV- and EBV+ donors. In HHV 8(+)/EBV+ LCLs, which have the phenotype of activated B lymphocytes (CD19(+), surface immunoglobulin M, CD23(+), CD30(+), CD80(+)), HHV-8 was still present after more than 25 passages (more than 9 months of culture). Latent viral transcripts and proteins were present in nonstimulated HHV-8(+)/EBV+ LCLs. After induction by phorbol ester and n-butyrate, HHV-8(+)/EBV+ LCLs expressed lytic HHV 8 transcripts and proteins. Moreover, HHV-8 could be serially passaged from HHV 8(+)/EBV+ LCLs to fresh PBMC. PMID- 9733856 TI - Development of a self-inactivating lentivirus vector. AB - We have constructed a new series of lentivirus vectors based on human immunodeficiency virus type 1 (HIV-1) that can transduce nondividing cells. The U3 region of the 5' long terminal repeat (LTR) in vector constructs was replaced with the cytomegalovirus (CMV) promoter, resulting in Tat-independent transcription but still maintaining high levels of expression. A self inactivating (SIN) vector was constructed by deleting 133 bp in the U3 region of the 3' LTR, including the TATA box and binding sites for transcription factors Sp1 and NF-kappaB. The deletion is transferred to the 5' LTR after reverse transcription and integration in infected cells, resulting in the transcriptional inactivation of the LTR in the proviruses. SIN viruses can be generated with no significant decreases in titer. Injection of viruses into the rat brain showed that a SIN vector containing the green fluorescent protein gene under the control of the internal CMV promoter transduced neurons as efficiently as a wild-type vector. Interestingly, a wild-type vector without an internal promoter also successfully transduced neurons in the brain, indicating that the HIV-1 LTR promoter is transcriptionally active in neurons even in the absence of Tat. Furthermore, injection of viruses into the subretinal space of the rat eye showed that wild-type vector transduced predominantly retinal pigment epithelium and photoreceptor cells, while SIN vector was able to transduce other types of retinal cells, including bipolar, Muller, horizontal, and amacrine cells. This finding suggests that the HIV-1 LTR can negatively influence the internal CMV promoter in some cell types. SIN HIV vectors should be safer for gene therapy, and they also have broader applicability as a means of high-level gene transfer and expression in nondividing cells. PMID- 9733857 TI - Functional analysis of the human cytomegalovirus US28 gene by insertion mutagenesis with the green fluorescent protein gene. AB - The protein encoded by the US28 gene of human cytomegalovirus (HCMV) has homology to G protein-coupled receptors (GCR). Previous studies demonstrated that recombinant US28 protein can bind the beta class of chemokines (K. Neote, D. DiGregorio, J. Y. Mak, R. Horuk, and T. J. Schall, Cell 72:415-425, 1993) and induce a rise in intracellular calcium after the binding of chemokines (J. L. Gao and P. M. Murphy, J. Biol. Chem. 269:28539-28542, 1994). In order to investigate the function of the US28 protein in virus-infected cells, a recombinant HCMV (HV5.8) was constructed, with the US28 open reading frame disrupted by the insertion of the Escherichia coli gpt gene and the gene for the green fluorescent protein. The US28 gene is not required for growth in human fibroblasts (HF). HF infected with wild-type HCMV bound RANTES at 24 h postinfection and demonstrated an intracellular calcium flux induced by RANTES. In cells infected with HV5.8, RANTES did not bind or induce a calcium flux, demonstrating that US28 is responsible for the beta-chemokine binding and induced calcium signaling in HCMV infected cells. The ability of the US28 gene to bind chemokines was shown to cause a significant reduction in the concentration of RANTES in the medium of infected cells. Northern analysis of RNA from infected cells showed that US28 is an early gene, while US27 (another GCR) is a late gene. PMID- 9733858 TI - A fifteen-amino-acid peptide inhibits human papillomavirus E1-E2 interaction and human papillomavirus DNA replication in vitro. AB - Mutation of the conserved glutamic acid residue at position 39 of human papillomavirus type 16 (HPV-16) E2 to alanine (E39A) disrupts its E1 interaction activity and its replication function in transient replication assays but does not affect E2 transcriptional activation. This E39A mutation also disrupts replication activity of HPV-16 E2 in HPV-16 in vitro DNA replication. On this basis, we designed 23- and 15-amino-acid peptides derived from HPV-16 E2 sequences flanking the E39 residue and tested the ability of these peptides to inhibit interaction between HPV-16 E1 and E2 in vitro. The inhibitory activity of these peptides was specific, since analogous peptides in which alanine was substituted for the E39 residue did not inhibit interaction. The 15-amino-acid peptide E2N-WP15 was the smallest peptide tested that effectively inhibited HPV 16 E1-E2 interaction. This peptide also inhibited in vitro replication of HPV-16 DNA. The efficacy of E2N-WP15 was not exclusive to HPV-16: this peptide also inhibited interaction of HPV-11 E1 with the E2 proteins of both HPV-11 and HPV-16 and inhibited in vitro replication with these same combinations of E1 and E2 proteins. These results provide further evidence that E1-E2 interaction is required for papillomavirus DNA replication and constitute the first demonstration that inhibition of this interaction is sufficient to prevent HPV DNA replication in vitro. PMID- 9733859 TI - Human immunodeficiency virus type 1 induction mediated by genistein is linked to cell cycle arrest in G2. AB - Protein tyrosine kinase (PTK) phosphorylation is involved in cellular proliferation and differentiation processes that are key factors for human immunodeficiency virus type 1 (HIV-1) regulation in infected monocytic cells. Short-term exposure of the chronically infected promyelocytic OM10 cell line with the PTK inhibitor genistein induced a dose-dependent increase in p24 antigen production in culture supernatants. This induction persisted in the presence of the reverse transcriptase inhibitor, zidovudine, and was associated with an increased transcription of HIV-1 multiply spliced and unspliced RNAs, suggesting a transcriptional mechanism targeting the integrated provirus. Genistein induced cell differentiation, apoptosis, and a G2 arrest in the OM10 cells. Cell differentiation and apoptosis were not directly involved in the observed increase in HIV-1 replication that was closely linked to genistein-induced G2 arrest. Alleviation of the G2 arrest by pentoxyfylline resulted in a concomitant reduction of HIV-1 to baseline replication. Additionally, by flow cytometry, a significant increase in the number of p24 antigen-expressing cells was observed in cells arrested in G2 compared to those located in G1 or S. Tyrosine kinase inhibition was found not to be essential for enhanced viral replication, which seemed to be related to two other properties of genistein, inhibition of topoisomerase II activity and inhibition of phosphotidylinositol turnover. These findings are consistent with the recent observation that HIV-1 Vpr induces viral replication through preventing proliferation of cells by arresting them in G2 of the cell cycle and strongly suggest that manipulation of the cell cycle plays an important role in HIV-1 pathogenesis. PMID- 9733860 TI - Purification and characterization of a cellular protein that binds to the downstream activation sequence of the strict late UL38 promoter of herpes simplex virus type 1. AB - Previous work on the strict late (gamma) UL38 promoter of herpes simplex virus type 1 identified three cis-acting elements required for wild-type levels of transcription: a TATA box at -31, a consensus mammalian initiator element at the transcription start site, and a downstream activation sequence (DAS) at +20 to +33. DAS is found in similar locations on several other late promoters, suggesting an important regulatory role in late gene expression. In this communication, we further characterize the interaction between DAS and a cellular protein which is found in both uninfected and infected nuclear extracts. This protein was purified from HeLa nuclear extracts and identified as the DNA binding component (Ku heterodimer) of DNA-dependent protein kinase (DNA-PK) by peptide mapping. Highly purified DNA-PK was able to stimulate UL38 transcription in vitro approximately 10-fold. DAS is similar in sequence to another element, nuclear regulatory element 1 (NRE1) of the glucocorticoid-responsive mouse mammary tumor virus long terminal repeat. NRE1 is known to specifically bind Ku in the absence of DNA ends. We demonstrated that NRE1 is able to substitute for DAS in the UL38 promoter to activate transcription as measured by in vitro transcription and in vivo during infection of tissue culture cells with recombinant virus. Also, we found that the binding of DNA-PK to DAS involves the bases demonstrated to be important in UL38 transcription and that the 70-kDa subunit of Ku binds to DAS. PMID- 9733861 TI - The human cytomegalovirus UL74 gene encodes the third component of the glycoprotein H-glycoprotein L-containing envelope complex. AB - The human cytomegalovirus (HCMV) gCIII envelope complex is composed of glycoprotein H (gH; gpUL75), glycoprotein L (gL; gpUL115), and a third, 125-kDa protein not related to gH or gL (M. T. Huber and T. Compton, J. Virol. 71:5391 5398, 1997; L. Li, J. A. Nelson, and W. J. Britt, J. Virol. 71:3090-3097, 1997). Glycosidase digestion analysis demonstrated that the 125-kDa protein was a glycoprotein containing ca. 60 kDa of N-linked oligosaccharides on a peptide backbone of 65 kDa or less. Based on these biochemical characteristics, two HCMV open reading frames, UL74 and TRL/IRL12, were identified as candidate genes for the 125-kDa glycoprotein. To identify the gene encoding the 125-kDa glycoprotein, we purified the gCIII complex, separated the components by sodium dodecyl sulfate polyacrylamide gel electrophoresis, and subjected gH and the 125-kDa glycoprotein to amino acid microsequence analysis. Microsequencing of an internal peptide derived from purified 125-kDa glycoprotein yielded the amino acid sequence LYVGPTK. A FASTA search revealed an exact match of this sequence to amino acids 188 to 195 of the predicted product of the candidate gene UL74, which we have designated glycoprotein O (gO). Anti-gO antibodies reacted in immunoblots with a protein species migrating at ca. 100 to 125 kDa in lysates of HCMV-infected cells and with 100- and 125-kDa protein species in purified virions. Anti-gO antibodies also immunoprecipitated the gCIII complex and recognized the 125-kDa glycoprotein component of the gCIII complex. Positional homologs of the UL74 gene were found in other betaherpesviruses, and comparisons of the predicted products of the UL74 homolog genes demonstrated a number of conserved biochemical features. PMID- 9733862 TI - Measles virus fusion protein is palmitoylated on transmembrane-intracytoplasmic cysteine residues which participate in cell fusion. AB - [3H]palmitic acid was metabolically incorporated into the viral fusion protein (F) of Edmonston or freshly isolated measles virus (MV) during infection of human lymphoid or Vero cells. The uncleaved precursor F0 and the F1 subunit from infected cells and extracellular virus were both labeled, indicating that palmitoylation can take place prior to F0 cleavage and that palmitoylated F protein was incorporated into virus particles. [3H]palmitic acid was released from F protein upon hydroxylamine or dithiothreitol treatment, indicating a thioester linkage. In cells transfected with the cloned MV F gene, in which the cysteines located in the intracytoplasmic and transmembrane domains (Cys 506, 518, 519, 520, and 524) were replaced by serine, a major reduction of [3H]palmitic acid incorporation was observed for F mutated at Cys 506 and, to a lesser extent, at Cys 518 and Cys 524. We also observed incorporation of [3H]palmitic acid in the F1 subunit of canine distemper virus F protein. Cell fusion induced by cotransfection of cells with MV F and H (hemagglutinin) genes was significantly reduced after replacement of Cys 506 or Cys 519 with serine in the MV F gene. Transfection with the F gene with a mutation for Cys 518 abolished cell fusion, although less mutant protein was detected on the cell surface. These results suggest that the F protein transmembrane domain cysteines 506 and 518 participate in structures involved in cell fusion, possibly mediated by palmitoylation. PMID- 9733864 TI - The RNA polymerase of influenza virus, bound to the 5' end of virion RNA, acts in cis to polyadenylate mRNA. AB - We previously demonstrated, by limited mutagenesis, that conserved sequence elements within the 5' end of influenza virus virion RNA (vRNA) are required for the polyadenylation of mRNA in vitro. To further characterize the nucleotide residues at the 5' end of vRNA which might be involved in polyadenylation, a complete set of short and long model vRNA-like templates with mutations at nucleotides 1' to 13' (prime notation denotes numbering from the 5' end) of vRNA were synthesized and transcribed in vitro. The products were assayed for mRNA production with both reverse transcription-PCR and [alpha-32P]ATP incorporation assays. Results from these independent assays showed that vRNA templates with point mutations at positions 2', 3', 7' to 9', and 11' to 13' synthesized polyadenylated transcripts inefficiently compared with those with mutations at positions 1', 4' to 6', and 10'. Positions 2', 3', 7' to 9', and 11' are known to be involved in RNA polymerase binding. Furthermore, residues at positions 11' to 13' are known to be involved in base pairing between the 3' and 5' ends of vRNA. These findings demonstrate that the RNA polymerase has to bind to the 5' end of the template vRNA, which must then interact with the 3' end of the same template for polyadenylation to occur. These results support a model in which a cis-acting RNA polymerase is required for the polyadenylation of influenza virus. PMID- 9733863 TI - Cleavage susceptibility of reovirus attachment protein sigma1 during proteolytic disassembly of virions is determined by a sequence polymorphism in the sigma1 neck. AB - A requisite step in reovirus infection of the murine intestine is proteolysis of outer-capsid proteins to yield infectious subvirion particles (ISVPs). When converted to ISVPs by intestinal proteases, virions of reovirus strain type 3 Dearing (T3D) lose 90% of their original infectivity due to cleavage of viral attachment protein sigma1. In an analysis of eight field isolate strains of type 3 reovirus, we identified one additional strain, type 3 clone 31 (T3C31), that loses infectivity and undergoes sigma1 cleavage upon conversion of virions to ISVPs. We examined the sigma1 deduced amino acid sequences of T3D and the eight field isolate strains for a correlation between sequence variability and sigma1 cleavage. The sigma1 proteins of T3D and T3C31 contain a threonine at amino acid position 249, whereas an isoleucine occurs at this position in the sigma1 proteins of the remaining strains. Thr249 occupies the d position of a heptad repeat motif predicted to stabilize sigma1 oligomers through alpha-helical coiled coil interactions. This region of sequence comprises a portion of the fibrous tail domain of sigma1 known as the neck. Substitution of Thr249 with isoleucine or leucine resulted in resistance to cleavage by trypsin, whereas replacement with asparagine did not affect cleavage susceptibility. These results demonstrate that amino acid position 249 is an independent determinant of T3D sigma1 cleavage susceptibility and that an intact heptad repeat is required to confer cleavage resistance. We performed amino-terminal sequence analysis on the sigma1 cleavage product released during trypsin treatment of T3D virions to generate ISVPs and found that trypsin cleaves sigma1 after Arg245. Thus, the sequence polymorphism at position 249 controls cleavage at a nearby site in the neck region. The relevance of these results to reovirus infection in vivo was assessed by treating virions with the contents of a murine intestinal wash under conditions that result in generation of ISVPs. The pattern of sigma1 cleavage susceptibility generated by using purified protease was reproduced in assays using the intestinal wash. These results provide a mechanistic explanation for sigma1 cleavage during exposure of virions to intestinal proteases and may account for certain strain-dependent patterns of reovirus pathogenesis. PMID- 9733866 TI - Identification of a negative cis element within the ZII domain of the Epstein Barr virus lytic switch BZLF1 gene promoter. AB - The Epstein-Barr virus (EBV) lytic switch gene, BZLF1, is tightly regulated in latently infected B cells. The BZLF1 gene promoter (Zp) contains several cis elements that have been previously shown to respond to inducers of the viral lytic cycle. These include four copies of an element referred to as the ZI domains and an element that contains a consensus CRE/AP-1 motif (ZII domain). In addition, Zp is autoregulated through two sites that bind the BZLF1 gene product Zta. The ZI domains have been shown to bind the ubiquitous cellular transcription factors Sp1 and Sp3 and/or the myocyte enhancer factor 2D (Liu et al., EMBO J. 16:143-153, 1997; Liu et al., Virology 228:9-16, 1997). Here we present a functional analysis of the ZII domain and show: (i) ATF-1 and ATF-2 appear to be the predominant cellular factors that bind to the CRE/AP-1 motif present in the ZII domain; and (ii) the region immediately upstream of the CRE/AP-1 motif contains a potent negative cis element, mutation of which results in a >10-fold increase in Zp activity. The negative cis element (ZIIR) in the ZII domain decreases both basal and induced Zp activity and thus is likely to play an important role in regulating reactivation of EBV. In addition, analysis of heterologous promoter constructs indicates that the function of ZIIR is context sensitive. Attempts to demonstrate a cellular factor binding to ZIIR have been unsuccessful, leaving unresolved the mechanism by which repression is mediated. PMID- 9733865 TI - Nasal immunization of mice with human papillomavirus type 16 virus-like particles elicits neutralizing antibodies in mucosal secretions. AB - To specifically induce a mucosal antibody response to purified human papillomavirus type 16 (HPV16) virus-like particles (VLP), we immunized female BALB/c mice orally, intranasally, and/or parenterally and evaluated cholera toxin (CT) as a mucosal adjuvant. Anti-HPV16 VLP immunoglobulin G (IgG) and IgA titers in serum, saliva, and genital secretions were measured by enzyme-linked immunosorbent assay (ELISA). Systemic immunizations alone induced HPV16 VLP specific IgG in serum and, to a lesser extent, in genital secretions but no secretory IgA. Oral immunization, even in the presence of CT, was inefficient. However, three nasal immunizations with 5 microgram of VLP given at weekly intervals to anesthetized mice induced high (>10(4)) and long-lasting (>15 weeks) titers of anti-HPV16 VLP antibodies in all samples, including IgA and IgG in saliva and genital secretions. CT enhanced the VLP-specific antibody response 10 fold in serum and to a lesser extent in saliva and genital secretions. Nasal immunization of conscious mice compared to anesthetized mice was inefficient and correlated with the absence of uptake of a marker into the lung. However, a 1 microgram VLP systemic priming followed by two 5-microgram VLP intranasal boosts in conscious mice induced both HPV16 VLP-specific IgG and IgA in secretions, although the titers were lower than in anesthetized mice given three intranasal immunizations. Antibodies in serum, saliva, and genital secretions of immunized mice were strongly neutralizing in vitro (50% neutralization with ELISA titers of 65 to 125). The mucosal and systemic/mucosal HPV16 VLP immunization protocols that induced significant titers of neutralizing IgG and secretory IgA in mucosal secretions in mice may be relevant to genital HPV VLP-based human vaccine trials. PMID- 9733867 TI - Evolution of envelope sequences from the genital tract and peripheral blood of women infected with clade A human immunodeficiency virus type 1. AB - The development of viral diversity during the course of human immunodeficiency virus type 1 (HIV-1) infection may significantly influence viral pathogenesis. The paradigm for HIV-1 evolution is based primarily on studies of male cohorts in which individuals were presumably infected with a single virus variant of subtype B HIV-1. In this study, we evaluated virus evolution based on sequence information of the V1, V2, and V3 portions of HIV-1 clade A envelope genes obtained from peripheral blood and cervical secretions of three women with genetically heterogeneous viral populations near seroconversion. At the first sample following seroconversion, the number of nonsynonymous substitutions per potential nonsynonymous site (dn) significantly exceeded substitutions at potential synonymous sites (ds) in plasma viral sequences from all individuals. Generally, values of dn remained higher than values of ds as sequences from blood or mucosa evolved. Mutations affected each of the three variable regions of the envelope gene differently; insertions and deletions dominated changes in V1, substitutions involving charged amino acids occurred in V2, and sequential replacement of amino acids over time at a small subset of positions distinguished V3. The relationship among envelope nucleotide sequences obtained from peripheral blood mononuclear cells, plasma, and cervical secretions was evaluated for each individual by both phylogenetic and phenetic analyses. In all subjects, sequences from within each tissue compartment were more closely related to each other than to sequences from other tissues (phylogenetic tissue compartmentalization). At time points after seroconversion in two individuals, there was also greater genetic identity among sequences from the same tissue compartment than among sequences from different tissue compartments (phenetic tissue compartmentalization). Over time, temporal phylogenetic and phenetic structure was detectable in mucosal and plasma viral samples from all three women, suggesting a continual process of migration of one or a few infected cells into each compartment followed by localized expansion and evolution of that population. PMID- 9733868 TI - Interaction of human immunodeficiency virus type 1 Tat with the transcriptional coactivators p300 and CREB binding protein. AB - Human immunodeficiency virus type 1 (HIV-1) encodes the transactivator protein Tat, which is essential for viral replication and progression to disease. Here we demonstrate that transcriptional activation by HIV-1 Tat involves p300 or the related cellular transcriptional coactivator CREB binding protein (CBP). Tat transactivation was inhibited by the 12S form of the adenovirus E1A gene product, which inhibits p300 function, and this inhibition was independent of its effect on NF-kappaB transcription. A biochemical interaction of p300 with Tat was demonstrated in vitro and in vivo by coimmunoprecipitation. The carboxy-terminal region of p300, which binds to E1A, was shown to bind specifically to the highly conserved basic domain of Tat, which also mediates binding to the Tat-responsive region RNA stem-loop structure. The ability of Tat to interact physically and functionally with this coactivator provides a mechanism to assemble a basal transcription complex which may subsequently respond to the effect of Tat on transcriptional elongation and represents a novel interaction between an RNA binding protein and a transcriptional coactivator. PMID- 9733869 TI - Herpes simplex virus type 1 glycoprotein gC mediates immune evasion in vivo. AB - Many microorganisms encode proteins that interact with molecules involved in host immunity; however, few of these molecules have been proven to promote immune evasion in vivo. Herpes simplex virus type 1 (HSV-1) glycoprotein C (gC) binds complement component C3 and inhibits complement-mediated virus neutralization and lysis of infected cells in vitro. To investigate the importance of the interaction between gC and C3 in vivo, we studied the virulence of a gC-null strain in complement-intact and C3-deficient animals. Using a vaginal infection model in complement-intact guinea pigs, we showed that gC-null virus grows to lower titers and produces less severe vaginitis than wild-type or gC rescued virus, indicating a role for gC in virulence. To determine the importance of complement, studies were performed with C3-deficient guinea pigs; the results demonstrated significant increases in vaginal titers of gC-null virus, while wild type and gC rescued viruses showed nonsignificant changes in titers. Similar findings were observed for mice where gC null virus produced significantly less disease than gC rescued virus at the skin inoculation site. Proof that C3 is important was provided by studies of C3 knockout mice, where disease scores of gC null virus were significantly higher than in complement-intact mice. The results indicate that gC-null virus is approximately 100-fold (2 log10) less virulent that wild-type virus in animals and that gC-C3 interactions are involved in pathogenesis. PMID- 9733870 TI - Induction of a mucosal cytotoxic T-lymphocyte response by intrarectal immunization with a replication-deficient recombinant vaccinia virus expressing human immunodeficiency virus 89.6 envelope protein. AB - To improve the safety of recombinant vaccinia virus vaccines, modified vaccinia virus Ankara (MVA) has been employed, because it has a replication defect in most mammalian cells. Here we apply MVA to human immunodeficiency virus type 1 (HIV-1) vaccine development by incorporating the envelope protein gp160 of HIV-1 primary isolate strain 89.6 (MVA 89.6) and use it to induce mucosal cytotoxic-T lymphocyte (CTL) immunity. In initial studies to define a dominant CTL epitope for HIV-1 89.6 gp160, we mapped the epitope to a sequence, IGPGRAFYAR (from the V3 loop), homologous to that recognized by HIV MN loop-specific CTL and showed that HIV-1 MN-specific CTLs cross-reactively recognize the corresponding epitope from strain 89.6 presented by H-2Dd. Having defined the CTL specificity, we immunized BALB/c mice intrarectally with recombinant MVA 89.6. A single mucosal immunization with MVA 89.6 was able to elicit long-lasting antigen-specific mucosal (Peyer's patch and lamina propria) and systemic (spleen) CTL responses as effective as or more effective than those of a replication-competent vaccinia virus expressing 89.6 gp160. Immunization with MVA 89.6 led to (i) the loading of antigen-presenting cells in vivo, as measured by the ex vivo active presentation of the P18-89.6 peptide to an antigen-specific CTL line, and (ii) the significant production of the proinflammatory cytokines (interleukin-6 and tumor necrosis factor alpha) in the mucosal sites. These results indicate that nonreplicating recombinant MVA may be at least as effective for mucosal immunization as replicating recombinant vaccinia virus. PMID- 9733871 TI - Naive and memory CD4 T cells differ in their susceptibilities to human immunodeficiency virus type 1 infection following CD28 costimulation: implicatip6s for transmission and pathogenesis. AB - In vitro evidence suggests that memory CD4(+) cells are preferentially infected by human immunodeficiency virus type 1 (HIV-1), yet studies of HIV-1-infected individuals have failed to detect preferential memory cell depletion. To explore this paradox, we stimulated CD45RA+ CD4(+) (naive) and CD45RO+ CD4(+) (memory) cells with antibodies to CD3 and CD28 and infected them with either CCR5 dependent (R5) or CXCR4-dependent (X4) HIV-1 isolates. Naive CD4(+) cells supported less X4 HIV replication than their memory counterparts. However, naive cells were susceptible to R5 viral infection, while memory cells remained resistant to infection and viral replication. As with the unseparated cells, mixing the naive and memory cells prior to infection resulted in cells resistant to R5 infection and highly susceptible to X4 infection. While both naive and memory CD4(+) subsets downregulated CCR5 expression in response to CD28 costimulation, only the memory cells produced high levels of the beta-chemokines RANTES, MIP-1alpha, and MIP-1beta upon stimulation. Neutralization of these beta chemokines rendered memory CD4(+) cells highly sensitive to infection with R5 HIV 1 isolates, indicating that downregulation of CCR5 is not sufficient to mediate complete protection from CCR5 strains of HIV-1. These results indicate that susceptibility to R5 HIV-1 isolates is determined not only by the level of CCR5 expression but also by the balance of CCR5 expression and beta-chemokine production. Furthermore, our results suggest a model of HIV-1 transmission and pathogenesis in which naive rather than memory CD4(+) T cells serve as the targets for early rounds of HIV-1 replication. PMID- 9733872 TI - Long-term CD4 Th1 and Th2 memory following acute lymphocytic choriomeningitis virus infection. AB - CD4 T cells play a central role in viral immunity. They provide help for B cells and CD8 T cells and can act as effectors themselves. Despite their importance, relatively little is known about the magnitude and duration of virus-specific CD4 T-cell responses. In particular, it is not known whether both CD4 Th1 memory and CD4 Th2 memory can be induced by viral infections. To address these issues, we quantitated virus-specific CD4 Th1 (interleukin 2 [IL-2] and gamma-interferon) and Th2 (IL-4) responses in mice acutely infected with lymphocytic choriomeningitis virus (LCMV). Using two sensitive assays (enzyme-linked immunospot assay and intracellular stain) to measure cytokine production at the single-cell level, we found that both CD4 Th1 and Th2 responses were induced during primary LCMV infection. At the peak (day 8) of the response, the frequency of LCMV-specific CD4 Th1 cells was 1/35 to 1/160 CD4 T cells, and the frequency of Th2 cells was 1/400. After viral clearance, the numbers of virus-specific CD4 T cells dropped to 1/260 to 1/3,700 and then were maintained at this level indefinitely. Upon rechallenge with LCMV, both CD4 Th1 and Th2 memory cells made an anamnestic response in vivo. These results show that unlike some microbial infections in which only Th1 or Th2 responses are seen, an acute viral infection can induce a mixed CD4 T-cell response with long-term memory. PMID- 9733873 TI - Structure and distribution of endogenous nonecotropic murine leukemia viruses in wild mice. AB - Virtually all of our present understanding of endogenous murine leukemia viruses (MLVs) is based on studies with inbred mice. To develop a better understanding of the interaction between endogenous retroviruses and their hosts, we have carried out a systematic investigation of endogenous nonecotropic MLVs in wild mice. Species studied included four major subspecies of Mus musculus (M. m. castaneus, M. m. musculus, M. m. molossinus, and M. m. domesticus) as well as four common inbred laboratory strains (AKR/J, HRS/J, C3H/HeJ, and C57BL/6J). We determined the detailed distribution of nonecotropic proviruses in the mice by using both env- and long terminal repeat (LTR)-derived oligonucleotide probes specific for the three different groups of endogenous MLVs. The analysis indicated that proviruses that react with all of the specific probes are present in most wild mouse DNAs tested, in numbers varying from 1 or 2 to more than 50. Although in common inbred laboratory strains the linkage of group-specific sequences in env and the LTR of the proviruses is strict, proviruses which combine env and the LTR sequences from different groups were commonly observed in the wild-mouse subspecies. The "recombinant" nonecotropic proviruses in the mouse genomes were amplified by PCR, and their genetic and recombinant natures were determined. These proviruses showed extended genetic variation and provide a valuable probe for study of the evolutionary relationship between MLVs and the murine hosts. PMID- 9733875 TI - A cluster of latently expressed genes in Kaposi's sarcoma-associated herpesvirus. AB - Infection with Kaposi's sarcoma-associated herpesvirus (KSHV) is closely associated with Kaposi's sarcoma (KS) and primary effusion lymphoma, with viral genomes present in a latent state in the majority of tumor cells. Here we describe a cluster of latently expressed viral genes whose mRNAs are generated from a common promoter. Two mRNAs in this region encode the latency-associated nuclear antigen, the product of open reading frame 73 (ORF73). The larger RNA, of 5.8 kb, is an unspliced transcript that includes ORF72 and -71 at its 3' end; it initiates at nucleotides (nt) 127880 to 127886 from a promoter lacking recognizable TATA elements. A less abundant mRNA, of 5.4 kb, is a variant of this transcript, in which 336 nt of 5' noncoding information has been removed by RNA splicing. A third, more abundant RNA is generated from the same promoter region via splicing from the common splice donor at nt 127813 to an acceptor 5' to ORF72; this transcript is the presumed mRNA for ORF72, which encodes the viral cyclin D homolog. All three RNAs are 3' coterminal. In situ hybridization analysis with probes that can detect all three transcripts shows that the RNAs are detectable in a large fraction of BCBL-1 cells prior to lytic induction and in >70% of KS spindle cells in primary KS tumors. This confirms that these transcripts are indeed latent RNAs and suggests a role for their products in viral persistence and/or KSHV-associated proliferation. PMID- 9733874 TI - Efficient class II major histocompatibility complex presentation of endogenously synthesized hepatitis C virus core protein by Epstein-Barr virus-transformed B lymphoblastoid cell lines to CD4(+) T cells. AB - The induction of an efficient CD4(+) T-cell response against hepatitis C virus (HCV) is critical for control of the chronicity of HCV infection. The ability of HCV structural protein endogenously expressed in an antigen-presenting cell (APC) to be presented by class II major histocompatibility complex molecules to CD4(+) T cells was investigated by in vitro culture analyses using HCV core-specific T cell lines and autologous Epstein-Barr virus-transformed B-lymphoblastoid cell lines (B-LCLs) expressing structural HCV antigens. The T- and B-cell lines were generated from peripheral blood mononuclear cells derived from HCV-infected patients. Expression and intracellular localization of core protein in transfected cells were determined by immunoblotting and immunofluorescence. By stimulation with autologous B-LCLs expressing viral antigens, strong T-cell proliferative responses were induced in two of three patients, while no substantial stimulatory effects were produced by B-LCLs expressing a control protein (chloramphenicol acetyltransferase) or by B-LCLs alone. The results showed that transfected B cells presented mainly endogenously synthesized core peptides. Presentation of secreted antigens from adjacent antigen-expressing cells was not enough to stimulate a core-specific T-cell response. Only weak T cell proliferative responses were generated by stimulation with B-LCLs that had been pulsed beforehand with at least a 10-fold-higher amount of transfected COS cells in the form of cell lysate, suggesting that presentation of antigens released from dead cells in the B-LCL cultures had a minimal role. Titrating numbers of APCs, we showed that as few as 10(4) transfected B-LCL APCs were sufficient to stimulate T cells. This presentation pathway was found to be leupeptin sensitive, and it can be blocked by antibody to HLA class II (DR). In addition, expression of a costimulatory signal by B7/BB1 on B cells was essential for T-cell activation. PMID- 9733876 TI - In vivo translation of the triple gene block of potato virus X requires two subgenomic mRNAs. AB - The 25-kilodalton (25K), 12K, and 8K movement proteins of potato virus X are derived from overlapping open reading frames (ORFs). Using an in vivo complementation assay, we have shown that the 25K protein is expressed from a functionally monocistronic mRNA, whereas the 12K and 8K proteins are from a bicistronic mRNA. Translation of the 8K ORF is by leaky ribosome scanning through the 12K ORF. PMID- 9733877 TI - Establishment and characterization of a human Epstein-Barr virus-associated gastric carcinoma in SCID mice. AB - A transplantable human Epstein-Barr virus-associated gastric carcinoma (EBVaGC), designated KT, was propagated in severe combined immunodeficiency (SCID) mice for 12 passages. Mucin and cytokeratin expression and the Alu sequence in tumor DNA confirmed that the KT tumor was derived from human epithelial tissue. The identity of clonal EBV in the original and KT tumors was demonstrated by terminal repeat analysis of EBV DNA. The pattern of latency gene expression of EBV was the same in both tumors. EBER1 was presented similarly in tumor cell nuclei by in situ hybridization. Reverse transcription-PCR analysis also demonstrated Q promoter-driven EBNA1 expression but not BZLF1, EBNA2, or LMP1 expression. Thus, the transplantable human EBVaGC KT retains the original EBV with the same latency gene expression and can serve as a model for this unique type of gastric carcinoma. PMID- 9733878 TI - Protection of mice against lethal coxsackievirus B3 infection by using DNA immunization. AB - Vaccination with DNA and recombinant vaccinia viruses (rec.VV) has been studied with the coxsackievirus B3 (CVB3) model system. Plasmids encoding all structural proteins of CVB3, when injected intramuscularly, induced only low levels of virus specific antibodies. However, DNA vaccination with the major structural protein VP1 protected 72.2% of mice from lethal challenge, whereas VP1 expressed by rec.VV was much less efficient. PMID- 9733879 TI - CREB-2, a cellular CRE-dependent transcription repressor, functions in association with Tax as an activator of the human T-cell leukemia virus type 1 promoter. AB - The Tax protein of the human T-cell leukemia virus type 1 (HTLV-1) has been implicated in human T-cell immortalization. The primary function of Tax is to transcriptionally activate the HTLV-1 promoter, but Tax is also known to stimulate expression of cellular genes. It has been reported to associate with several transcription factors, as well as proteins not involved in transcription. To better characterize potential cellular targets of Tax present in infected cells, a Saccharomyces cerevisiae two-hybrid screening was performed with a cDNA library constructed from the HTLV-1-infected MT2 cell line. From this study, we found 158 positive clones representing seven different cDNAs. We focused our attention on the cDNA encoding the transcription factor CREB-2. CREB-2 is an unconventional member of the ATF/CREB family in that it lacks a protein kinase A (PKA) phosphorylation site and has been reported to negatively regulate transcription from the cyclic AMP response element of the human enkephalin promoter. In this study, we demonstrate that CREB-2 cooperates with Tax to enhance viral transcription and that its basic-leucine zipper C-terminal domain is required for both in vitro and in vivo interactions with Tax. Our results confirm that the activation of the HTLV-1 promoter through Tax and factors of the ATF/CREB family is PKA independent. PMID- 9733880 TI - A functionally distinct TATA box required for late progression through the Epstein-Barr virus life cycle. AB - During EBV infection, lytic DNA replication activates late gene expression in trans via an uncharacterized pathway. In this study, we mapped the target of this regulatory cascade to a variant TATA box (TATTAAA) and the 3' flanking region within the core promoter of the BcLF1 gene. The inherent late activity of this core promoter is, surprisingly, disrupted by a heterologous enhancer, suggesting that late gene expression is regulated through core promoter sequences located in a transcriptionally inert environment. PMID- 9733881 TI - The S2 gene of equine infectious anemia virus is dispensable for viral replication in vitro. AB - Equine infectious anemia virus (EIAV) contains the simplest genome among lentiviruses in that it encodes only three putative regulatory genes (S1, S2, S3) in addition to the canonical gag, pol, and env genes, presumably reflecting its limited tropism to cells of monocyte/macrophage lineage. Tat and Rev functions have been assigned to S1 and S3, respectively, but the specific function for the S2 gene has yet to be determined. Thus, the function of S2 in virus replication in vitro was investigated by using an infectious molecular viral clone, EIAVUK. Various EIAVUK mutants lacking S2 were constructed, and their replication kinetics were examined in several equine cell culture systems, including the natural in vivo target equine macrophage cells. The EIAV S2 mutants showed replication kinetics similar to those of the parental virus in all of the tested primary and transformed equine cell cultures, without any detectable reversion of mutant genomes. The EIAVUK mutants also showed replication kinetics similar to those of the parental virus in an equine blood monocyte differentiation maturation system. These results demonstrate for the first time that the EIAV S2 gene is not essential and does not appear to affect virus infection and replication properties in target cells in vitro. PMID- 9733882 TI - Monitoring retroviral RNA dimerization in vivo via hammerhead ribozyme cleavage. AB - We have used a strategy for colocalization of Psi (Psi)-tethered ribozymes and targets to demonstrate that Psi sequences are capable of specific interaction in the cytoplasm of both packaging and nonpackaging cells. These results indicate that current in vitro dimerization models may have in vivo counterparts. The methodology used may be applied to further genetic analyses on Psi domain interactions in vivo. PMID- 9733883 TI - The reovirus protein mu2, encoded by the M1 gene, is an RNA-binding protein. AB - The reovirus M1, L1, and L2 genes encode proteins found at each vertex of the viral core and are likely to form a structural unit involved in RNA synthesis. Genetic analyses have implicated the M1 gene in viral RNA synthesis and core nucleoside triphosphatase activity, but there have been no direct biochemical studies of mu2 function. Here, we expressed mu2 in vitro and assessed its RNA binding activity. The expressed mu2 binds both poly(I-C)- and poly(U)-Sepharose, and binding activity is greater in Mn2+ than in Mg2+. Heterologous RNA competes for mu2 binding to reovirus RNA transcripts as effectively as homologous reovirus RNA does, providing no evidence for sequence-specific RNA binding by mu2. Protein mu2 is now the sixth reovirus protein demonstrated to have RNA-binding activity. PMID- 9733884 TI - Two novel adenovirus vector systems permitting regulated protein expression in gene transfer experiments. AB - Two new adenovirus vector systems based on the tetracycline-regulated Tet-ON- (Gossen, M., et al., Science 268:1766-1769, 1995) and the RU 486-regulated progesterone antagonist (Wang, Y., et al., Proc. Natl. Acad. Sci. USA 91:8180 8184, 1994)-induced gene expression systems are described. We show that both systems permit a tight control of chloramphenicol acetyltransferase reporter gene expression in a variety of cell types, with induction levels of approximately 1,800-fold (Tet-ON system) and 600-fold (RU 486-regulated system), respectively. A significant advantage of our vector systems is that reporter protein expression can be adjusted over a wide range by varying the amount of inducer. The Tet-ON system is also shown to permit an efficient control of reporter gene expression in mice. PMID- 9733885 TI - Mx1-based resistance to thogoto virus in A2G mice is bypassed in tick-mediated virus delivery. AB - The interferon-induced mouse Mx1 protein has intrinsic antiviral activity against orthomyxoviruses, including Thogoto virus. Thus, Mx1(+) A2G mice are apparently resistant to infection following needle- or tick-borne virus challenge. However, tick-borne challenge and, to a lesser degree, injection of virus mixed with tick salivary gland extract resulted in virus transmission to uninfected ticks feeding on the A2G mice. The data indicate that immunomodulatory components in tick saliva can overcome a natural antiviral mechanism. PMID- 9733886 TI - Location-specific, unequal contribution of the N glycans in simian immunodeficiency virus gp120 to viral infectivity and removal of multiple glycans without disturbing infectivity. AB - One of the striking features of human immunodeficiency virus, simian immunodeficiency virus (SIV), and other lentiviruses is extensive N glycosylation of the envelope protein. To assess the requirement of each N glycan for viral infectivity, we individually silenced all 23 N glycosylation sites in the gp120 subunit of SIVmac239 envelope protein by mutagenizing the canonical Asn-Xaa Thr/Ser N glycosylation motif in an infectious molecular clone, attempted to rescue viruses from the clones, and compared the replication capability of the rescued viruses in MT4 cells. The mutation resulted in either the recovery of a fully infectious virus (category I); recovery of a faster-replicating virus, compared with the parental virus (category II); or no virus recovery (category III). These categorically different sites were not distributed randomly but were clustered. The sites of category I were localized largely in the N-terminal half, whereas the sites of categories II and III were localized in the C-terminal region, including the CD4 binding site, and the central part, including the C loop, respectively. To learn how far SIV can tolerate the removal of glycans, multiplex mutagenesis was also attempted. When they were appreciably distant from one another in the primary sequence, up to five sites could be silenced in combination without disturbing infectivity. On the other hand, it was difficult to silence contiguous sites. Thus, it appeared that a certain degree of sugar chain density over the local region had to be preserved. We discuss the potential utility of these variously deglycosylated mutants for clarifying the role of N glycans in SIV replication in vivo, as well as in the host response, and for designing vaccines and the generation of glycoprotein crystals. PMID- 9733887 TI - An adenovirus type 5 mutant with the preterminal protein gene deleted efficiently provides helper functions for the production of recombinant adeno-associated virus. AB - Production of recombinant adeno-associated virus (rAAV) requires helper functions that have routinely been provided by infection of the producer cells with adenovirus. Complete removal and/or inactivation of progeny adenovirus, present in such rAAV preparations, presents significant difficulty. Here, we report that an adenovirus type 5 (Ad5) mutant with the preterminal protein (pTP) gene deleted can provide helper function for the growth of rAAV. At high multiplicity, Ad5dl308DeltapTP was as efficient as the phenotypically wild-type Ad5dl309 in permitting growth of rAAV. Use of Ad5dl308DeltapTP, which is incapable of replication in the absence of complementation for pTP, as a helper avoids the need to remove contaminating adenovirus infectious activity by heat inactivation or by purification. Comparison of the transducing ability of rAAV generated with either Ad5dl308DeltapTP or Ad5dl309 as a helper demonstrated that the heat inactivation protocol generally used does not remove all of the helper Ad5dl309 function. PMID- 9733888 TI - Cell surface proteoglycans are necessary for A27L protein-mediated cell fusion: identification of the N-terminal region of A27L protein as the glycosaminoglycan binding domain. AB - We previously showed that vaccinia virus infection of BSC40 cells was blocked by soluble heparin, suggesting that cell surface heparan sulfate mediates vaccinia virus binding (C.-S. Chung, J.-C. Hsiao, Y. -S. Chang, and W. Chang, J. Virol. 72:1577-1585, 1998). In this study, we extended our previous work and demonstrated that soluble A27L protein bound to heparan sulfate on cells and interfered with vaccinia virus infection at a postbinding step. In addition, we investigated the structure of A27L protein that provides for its binding to heparan sulfate on cells. A mutant of A27L protein, named D-A27L, devoid of a cluster of 12 amino acids rich in basic residues, was constructed. In contrast to the soluble A27L protein, purified D-A27L protein was inactive in all of our assays, including binding to heparin in vitro, binding to heparan sulfate on cells, and the ability to block virus infection. These data demonstrated that the N-terminal region acts as a glycosaminoglycan (GAG)-binding domain critical for A27L protein binding to cells. Previously A27L protein was thought to be involved in fusion of virus-infected cells induced by acid treatment. When we investigated whether cell surface GAGs also participate in A27L-dependent fusion, our results indicated that soluble A27L protein blocked cell fusion, whereas D-A27L protein did not. Taken together, the results therefore demonstrated that A27L-mediated cell fusion is triggered by its interaction with cell surface GAGs through the N terminal domain. PMID- 9733889 TI - 1,25-Dihydroxyvitamin D3 upregulates functional CXCR4 human immunodeficiency virus type 1 coreceptors in U937 minus clones: NF-kappaB-independent enhancement of viral replication. AB - U937 cell clones which sustain efficient or poor replication of human immunodeficiency virus type 1 (HIV-1) (referred to herein as plus clones and minus clones, respectively) have been previously described. 1,25-Dihydroxyvitamin D3 (vitamin D3) potently induced HIV-1 replication and proviral DNA accumulation in minus clones but not in plus clones. Vitamin D3 did not induce NF-kappaB activation but selectively upregulated CXCR4 expression in minus clones. The CXCR4 ligand stromal-cell derived factor-1 induced Ca2+ fluxes and inhibited both constitutive and vitamin D3-enhanced HIV replication in minus clones. PMID- 9733890 TI - Proviral structure, chromosomal location, and expression of HERV-K-T47D, a novel human endogenous retrovirus derived from T47D particles. AB - We previously described that type B retrovirus-like particles released from the human mammary carcinoma cell line T47D are pseudotypes and package retroviral RNA of different origins (W. Seifarth, H. Skladny, F. Krieg-Schneider, A. Reichert, R. Hehlmann, and C. Leib-Mosch, J. Virol. 69:6408-6416, 1995). One preferentially packaged retroviral sequence, ERV-MLN, has now been used to isolate the corresponding full-length provirus from a human genomic library. The 9,315-bp proviral genome comprises a complete retroviral structure except for a 3' long terminal repeat (LTR) truncation. A lysine tRNA primer-binding site and phylogenetic analyses assign this human endogenous retroviral element, now called HERV-K-T47D, to the HML-4 subgroup of the HERV-K superfamily. The gag, prt, pol, and env genes exhibit 40 to 60% amino acid identity to HERV-K10. HERV-K-T47D is located on human chromosome 10, with five closely related elements on chromosomes 8, 9, 15, 16, and 19 and several hundred HERV-K-T47D-related solitary LTRs dispersed over the human genome. HERV-K-T47D-related sequences are detected in the genomes of higher primates and Old World monkeys but not in those of New World monkeys. High HERV-K-T47D transcription levels were observed in human placenta tissue, whereas transcription in T47D cells was strictly steroid dependent. PMID- 9733891 TI - Suppression of c-Myc-induced apoptosis by the Epstein-Barr virus gene product BHRF1. AB - Constitutive expression of the c-myc proto-oncogene in growth factor-deprived fibroblasts promotes proliferation and induces apoptosis. In these cells, apoptosis can be inhibited by survival factors such as insulin-like growth factor I or the bcl-2 proto-oncogene product. Deregulated c-Myc expression is a common feature in Epstein-Barr virus-positive Burkitt's lymphoma in which the c-myc gene is reciprocally translocated and placed under the control of one of the immunoglobulin loci. BHRF1 is an Epstein-Barr virus protein expressed early in the lytic cycle. BHRF1 is a member of the Bcl-2 family and has been shown to suppress apoptosis and to increase cell survival in different settings. In the present study, we report that BHRF1 inhibits c-Myc-induced apoptosis which occurs in the absence of survival factors. It does not, however, affect the capacity of c-Myc to promote cell growth. These findings demonstrate that BHRF1 has not only structural but also functional similarities to Bcl-2. PMID- 9733892 TI - Specific and independent recognition of U3 and U5 att sites by human immunodeficiency virus type 1 integrase in vivo. AB - The retroviral attachment (att) sites at viral DNA ends are cis-acting regions essential for proviral integration. To investigate the sequence features of att important for human immunodeficiency virus type 1 (HIV-1) integration in vivo, we generated a series of 25 att mutants of HIV-1 by mutagenesis of the U3, U5, or both boundaries of att. Our results indicated that the terminal 11 or 12 bp of viral DNA are sufficient for specific recognition by HIV-1 integrase (IN) and suggested that IN might recognize each att site independently in vivo. PMID- 9733893 TI - Strain-specific differences in LFA-1 induction on measles virus-infected monocytes and adhesion and viral transmission to endothelial cells. AB - Measles virus (MV) infection of monocytes induces leukocyte function-associated antigen-1 (LFA-1), an integrin that mediates intercellular adhesion to the endothelium. Thus, an increase in LFA-1 expression could lead to enhanced monocyte adherence and virus dissemination to endothelial cells (ECs) and potentially be an important means of distinction between MV strains. We identified both vaccine and wild-type strains that induced LFA-1 and others that failed to induce. Although adhesion of MV-infected monocytes and viral transmission to ECs was demonstrated, strain-specific differences were not correlated with LFA-1 induction. MV infection of ECs was dramatically reduced in the absence of cell contact, suggesting virus dissemination by cell-cell transmission. PMID- 9733894 TI - Complete nucleotide sequence and genetic organization of Aichi virus, a distinct member of the Picornaviridae associated with acute gastroenteritis in humans. AB - The complete nucleotide sequence of a novel enteric virus, Aichi virus, associated with nonbacterial acute gastroenteritis in humans was determined. The Aichi virus genome proved to be a single-stranded positive-sense RNA molecule with 8,251 bases excluding a poly(A) tail; it contains a large open reading frame with 7,302 nucleotides that encodes a potential polyprotein precursor of 2,433 amino acids. The genome contains a 5' nontranslated region (NTR) with 712 bases and a 3' NTR with 240 bases followed by a poly(A) tail. The structure of the genome, VPg-5' NTR-leader protein-structural proteins-nonstructural proteins-3' NTR-poly(A), was found to be typical of a picornavirus. The VP0-VP3 and VP3-VP1 cleavage sites were determined to be Q-H and Q-T, respectively, by N-terminal amino acid sequence analyses using purified virion proteins. Possible cleavage sites, Q-G, Q-A, and Q-S, which cleave P2 and P3 polyproteins were found to be similar to those of picornaviruses. A dendrogram based on 3Dpol proteins indicated that Aichi virus is genetically distinct from the known six genera of picornaviruses including entero-, rhino-, cardio-, aphtho-, and hepatovirus and echovirus 22. Considering this together with other properties of the virus (T. Yamashita, S. Kobayashi, K. Sakae, S. Nakata, S. Chiba, Y. Ishihara, and S. Isomura, J. Infect. Dis. 164:954-957, 1991), we propose that Aichi virus be regarded as a new genus of the family Picornaviridae. PMID- 9733895 TI - Effect of vesicular stomatitis virus matrix protein on transcription directed by host RNA polymerases I, II, and III. AB - The matrix (M) protein of vesicular stomatitis virus (VSV) functions in virus assembly and inhibits host-directed gene expression independently of other viral components. Experiments in this study were carried out to determine the ability of M protein to inhibit transcription directed by each of the three host RNA polymerases (RNA polymerase I [RNAPI], RNAPII, and RNAPIII). The effects of wild type (wt) VSV, v6 (a VSV mutant isolated from persistently infected cells), and tsO82 viruses on poly(A)+ and poly(A)- RNA synthesis were measured by incorporation of [3H]uridine. v6 and tsO82 viruses, which contain M-gene mutations, had a decreased ability to inhibit synthesis of both poly(A)+ and poly(A)- RNA. Nuclear runoff analysis showed that VSV inhibited transcription of 18S rRNA and alpha-tubulin genes, which was dependent on RNAPI and RNAPII, respectively, but infection with wt virus enhanced transcription of 5S rRNA by RNAPIII. The effect of M protein alone on transcription by RNAPI-, RNAPII-, and RNAPIII-dependent promoters was measured by cotransfection assays. M protein inhibited transcription from RNAPI- and RNAPII-dependent promoters in the absence of other viral gene products. RNAPIII-dependent transcription of the adenovirus VA promoters was also inhibited by M protein. However, as observed during wt VSV infection, M protein enhanced endogenous 5S rRNA transcription, indicating that the inhibition of transcription by RNAPIII was dependent on the nature of the promoter. PMID- 9733897 TI - The major open reading frame of the beta2.7 transcript of human cytomegalovirus: in vitro expression of a protein posttranscriptionally regulated by the 5' region. AB - beta2.7 is the major early transcript produced during human cytomegalovirus infection. This abundantly expressed RNA is polysome associated, but no protein product has ever been detected. In this study, a stable peptide of 24 kDa was produced in vitro from the major open reading frame (ORF), TRL4. Following transient transfection, the intracellular localization was nucleolar and the expression was posttranscriptionally inhibited by the 5' sequence of the transcript, which harbors two short upstream ORFs. PMID- 9733896 TI - Resistance to the anti-human immunodeficiency virus type 1 compound L-chicoric acid results from a single mutation at amino acid 140 of integrase. AB - L-Chicoric acid is an inhibitor of human immunodeficiency virus type 1 (HIV-1) integrase in vitro and of HIV-1 replication in tissue culture. Following 3 months of selection in the presence of increasing concentrations of L-chicoric acid, HIV 1 was completely resistant to the compound. Introduction of the mutant integrase containing a single glycine-to-serine amino acid change at position 140 into the native, L-chicoric acid-sensitive virus demonstrated that this change was sufficient to confer resistance to L-chicoric acid. These results confirm through natural selection previous biochemical studies showing that L-chicoric acid inhibits integrase and that the drug is likely to interact at residues near the catalytic triad in the integrase active site. PMID- 9733898 TI - Optimal induction of hepatitis C virus envelope-specific immunity by bicistronic plasmid DNA inoculation with the granulocyte-macrophage colony-stimulating factor gene. AB - In this study, we have constructed various DNA vaccine vectors that carried hepatitis C virus (HCV) envelope genes without and with the granulocyte macrophage colony-stimulating factor (GM-CSF) gene in several different ways. In Buffalo rats that received plasmids carrying the HCV envelope genes, which encode envelope proteins E1 and E2, both antibody and lymphoproliferative responses against these proteins were induced. These responses were greatly enhanced by the codelivery of the GM-CSF gene. In particular, inoculation with a bicistronic plasmid that independently expressed the GM-CSF gene and the envelope genes in the same construct generated the highest antibody titers and significantly increased lymphoproliferative responses against these proteins. Moreover, strong antibody responses to homologous and heterologous hypervariable region 1 peptides were elicited in the immunized rats. PMID- 9733899 TI - Characterization of simian-human immunodeficiency virus envelope glycoprotein epitopes recognized by neutralizing antibodies from infected monkeys. AB - We characterized human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein epitopes recognized by neutralizing antibodies from monkeys recently infected by molecularly cloned simian-human immunodeficiency virus (SHIV) variants. The early neutralizing antibody response in each infected animal was directed mainly against a single epitope. This primary neutralizing epitope, however, differed among individual monkeys infected by identical viruses. Two such neutralization epitopes were determined by sequences in the V2 and V3 loops of the gp120 envelope glycoprotein, while a third neutralization epitope, apparently discontinuous, was determined by both V2 and V3 sequences. These results indicate that the early neutralizing antibody response in SHIV-infected monkeys is monospecific and directed against epitopes composed of the gp120 V2 and V3 variable loops. PMID- 9733901 TI - CXCR4 as a functional coreceptor for human immunodeficiency virus type 1 infection of primary macrophages. AB - The coreceptors used by primary syncytium-inducing (SI) human immunodeficiency virus type 1 isolates for infection of primary macrophages were investigated. SI strains using only CXCR4 replicated equally well in macrophages with or without CCR5 and were inhibited by several different ligands for CXCR4 including SDF-1 and bicyclam derivative AMD3100. SI strains that used a broad range of coreceptors including CCR3, CCR5, CCR8, CXCR4, and BONZO infected CCR5-deficient macrophages about 10-fold less efficiently than CCR5(+) macrophages. Moreover, AMD3100 blocked infection of CCR5-negative macrophages by these strains. Our results therefore demonstrate that CXCR4, as well as CCR5, is used for infection of primary macrophages but provide no evidence for the use of alternative coreceptors. PMID- 9733902 TI - Fluorescence quenching data interpretation in biological systems. The use of microscopic models for data analysis and interpretation of complex systems. AB - In micro-heterogeneous media (e.g. membranes, micelles and colloidal systems), the fluorescence decay in the absence of quencher is usually intrinsically complex, e.g. due to the existence of several sub-populations with different micro-environments. In this case it is impossible to analyze data in detail (accounting for transient effects) and simpler formalisms are needed. The objective of the present work is to present and discuss such simpler formalisms. The goal is to achieve simple data analysis and meaningful, clear data interpretation in complex systems using microscopic models that consider several sub-populations of chromophores. Two points are dealt with in detail. (i) It is shown that the approximation of the transient effects by the quenching sphere-of action model is not always possible. The quenching sphere-of-action concept can be regarded as a valuable tool, although crude, only in a limited range of experimental conditions, namely time resolution. (ii) The Stern-Volmer equation usually used for data analysis is only valid for a limited range of small and moderate equilibrium association constants, Ka, although this is frequently overlooked in the literature. Self-consistency criteria are presented for the proposed methods. The well-known downward curvature due to a fraction of fluorophores which is not accessible to the quencher is only a limiting case from a set of possible situations which result in deviations to linearity. A systematic classification of the different types of quenching is presented. PMID- 9733900 TI - Dysregulation through the NF-kappaB enhancer and TATA box of the human immunodeficiency virus type 1 subtype E promoter. AB - The global diversity of human immunodeficiency virus type 1 (HIV-1) genotypes, termed subtypes A to J, is considerable and growing. However, relatively few studies have provided evidence for an associated phenotypic divergence. Recently, we demonstrated subtype-specific functional differences within the long terminal repeat (LTR) region of expanding subtypes (M. A. Montano, V. A. Novitsky, J. T. Blackard, N. L. Cho, D. A. Katzenstein, and M. Essex, J. Virol. 71:8657-8665, 1997). Notably, all HIV-1E isolates were observed to contain a defective upstream NF-kappaB site and a unique TATA-TAR region. In this study, we demonstrate that tumor necrosis factor alpha (TNF-alpha) stimulation of the HIV-1E LTR was also impaired, consistent with a defective upstream NF-kappaB site. Furthermore, repair of the upstream NF-kappaB site within HIV-1E partially restored TNF-alpha responsiveness. We also show, in gel shift assays, that oligonucleotides spanning the HIV-1E TATA box displayed a reduced efficiency in the assembly of the TBP TFIIB-TATA complex, relative to an HIV-1B TATA oligonucleotide. In transfection assays, the HIV-1E TATA, when changed to the canonical HIV-1B TATA sequence (ATAAAA-->ATATAA) unexpectedly reduces both heterologous HIV-1B Tat and cognate HIV-1E Tat activation of an HIV-1E LTR-driven reporter gene. However, Tat activation, irrespective of subtype, could be rescued by introducing a cognate HIV-1B TAR. Collectively, these observations suggest that the expanding HIV-1E genotype has likely evolved an alternative promoter configuration with altered NF kappaB and TATA regulatory signals in contradistinction with HIV-1B. PMID- 9733903 TI - Melanin-concentrating hormone and neuropeptide EI projections from the lateral hypothalamic area and zona incerta to the medial septal nucleus and spinal cord: a study using multiple neuronal tracers. AB - The projection pathways of neurons containing melanin-concentrating hormone (MCH) and neuropeptide EI (NEI), two peptides colocalized in the lateral hypothalamic area (LHA) of the rat, were mapped using the retrogradely transported fluorescent dyes, true blue (TB) and diamidino yellow (DY). TB and DY were injected into the medial septum/diagonal band complex (MS/DBC) and the thoracic level of the spinal cord (SpCd), respectively. Brains from rats receiving only one or both tracer injections were immunohistochemically stained for MCH in the spinal cord and NEI in the forebrain. In the MS/DBC, NEI-immunoreactive (-ir) fibers are concentrated in the MS and in the vertical and horizontal limbs of the DBC. In the SpCd, MCH ir fibers are concentrated primarily in lamina X. Of the diencephalic NEI-ir neurons, 37.15% project to the MS/DBC and reside in the rostromedial zona incerta (ZIm), in the LHAt and LHAp, and in the perifornical region. Of the diencephalic MCH-ir neurons, 20.2% project to the SpCd and reside in the LHAt and LHAp. In addition, 2. 2% of the MCH-ir cells and 8.7% of the NEI-ir cells in the hypothalamus were labeled with both retrograde tracers and thus project to both the MS/DBC and SpCd. These dual projection neurons are located mainly in the LHAt and LHAp. Anterograde injections of the tracer Phaseolus vulgaris leucoagglutinin into the LHAt and ZIm corroborated our findings in the retrograde studies. Potential autonomic and behavioral roles of the NEI and MCH systems in the MS/DBC and the SpCd are discussed. PMID- 9733904 TI - Expression of neurotrophins and neurotrophin receptors in the cerebellum of mutant weaver and lurcher mice. AB - To test the hypothesis whether a failure to express neurotrophins or a neurotrophin receptor might underlie the pathology observed in mutant mice with degeneration of regionally distinct subpopulations of neurons, the expression of BDNF, NT-3, TrkB, TrkC and synaptophysin mRNA was examined in the cerebellum of mutant lurcher (lc/+) and weaver (wv/+)/(wv/wv) mice. To identify the expression patterns of individual neurons, we used in situ hybridization with digoxigenin labeled ribonucleotide probes. RT-PCR of cerebellar mRNA for BDNF, NT-3, TrkB and TrkC (GAPDH as internal standard) was performed in parallel. Although especially in homozygous (wv/wv) weaver mice the normal anatomical order and number of the cerebellar neurons is grossly disturbed, residual Purkinje and granule neurons of both mutants displayed a normal expression pattern of the neurotrophins examined. Thus, the affected animals showed no significant signal decrease compared to healthy littermates or C3H mice. Our results suggest that the loss of specific neuron populations in the cerebellum of either mutant occurs via mechanisms either independent or downstream of the neurotrophins examined in this study. PMID- 9733905 TI - Appearance and distribution of two Ca2+-binding proteins during development of the cochlea in the musk shrew. AB - In the developing cochlea of the musk shrew, Suncus murinus, the localization of two Ca2+-binding protein, calbindin and calmodulin, which are thought to play different roles in the nervous system, was examined during gestational and postpartum periods. Calbindin is thought to play a Ca2+ buffering role, while calmodulin activates other proteins. Cochleae from the musk shrews sacrificed from gestational day (GD) 15 to postnatal day (PP) 9 and as adults, were immunohistochemically analyzed. The localization and order of appearance of calmodulin in sensorineural elements were similar to those of calbindin, except for timing of appearance. Calmodulin-staining was recognized first in the spiral ganglion neurons on GD21, followed by the inner hair cells (IHCs) on GD23 and outer hair cells (OHCs) on GD26, while calbindin immunoreactivity in the spiral ganglion neurons on GD19, the IHCs on GD21 and the OHCs on GD23. In hair cells, during development, immunostaining of calbindin and calmodulin was initially seen in the cytoplasm, followed by the cuticular plate. Cytoplasmic staining then decreased in mature hair cells. Non-sensorineural components also showed positivity for both calbindin and calmodulin. The lateral wall of the cochlear duct was positive for calbindin, while the stria vascularis was positive for calmodulin. Immunoreactivity for calbindin was present earlier than that of calmodulin in sensorineural elements, suggesting that in the developing cochlea, calbindin and calmodulin have different functions and that Ca2+ buffering capacity, which is regulated by Ca2+ buffer proteins, such as calbindin, may be required before trigger proteins, such as calmodulin, function. PMID- 9733906 TI - Lysosome membrane permeability to anions. AB - The permeability of rat liver lysosomes to some inorganic and aliphatic organic anions was investigated, using an osmotic-protection methodology. Lysosomes were incubated at 25 degreesC in 250 mOsm solutions of potassium salts of the anions, in the presence of valinomycin, and the latency of lysosomal hexosaminidase measured at intervals. Lysosomes suspended in 250 mM sucrose at 25 degreesC were stable for up to 4 h. When suspended in 250 mOsm solutions of potassium salts of inorganic acids, latency was lost at rates indicating anion permeance decreasing in the order thiocyanate, nitrate and iodide>bromide>chloride>sulfate. This rank order does not correspond with the anion selectivity of any known anion transporter, and is closer to that of the lyotropic series. Results with the potassium salts of aliphatic organic acids indicate little correlation between permeation and hydrocarbon chain length, although formate was more rapidly permeant than acetate and its higher homologs. By contrast, oxalate was less permeable than other dicarboxylic acids. The presence of one or more hydroxy groups decreased permeance. A correlation between permeance and the acid's lowest pKa suggested that penetration was due principally to the entry of the undissociated acid, but there is evidence that the (much more abundant) singly charged anionic form is also significantly permeant. PMID- 9733907 TI - The effect of hypothalamic peptide YY on hippocampal acetylcholine release in vivo: implications for limbic function in binge-eating behavior. AB - Central injection of peptide YY (PYY) in sated rats produces the most powerful stimulating effect of food intake known to date. The neural mechanisms by which PYY regulates appetite are not clear but may be important because abnormal levels of PYY have been implicated in the neurobiology of bulimia nervosa. Interactions between brain acetylcholine (ACh) and PYY had not been studied. Therefore, the present experiments were designed to explore the in vivo release of ACh from the hippocampus (HPC) of rats in response to hypothalamic infusion of PYY. Hippocampal ACh release was found to increase 400% in response to 10 microg PYY. In a separate experiment, blockade of the same area of the HPC with bilateral intracerebral injections of 3.5 microg scopolamine did not affect intake stimulated by intrahypothalamic injection of 4 microg PYY. Furthermore, a third experiment showed, for the first time, that PYY (2.5-10.0 microg) can elicit robust feeding when infused directly into the HPC. The significance of these findings to the activation of limbic functions such as memory, reinforcement, and obsessional processes that accompany human binge-eating syndromes is discussed. PMID- 9733908 TI - Localization of mRNAs for CDP-diacylglycerol synthase and phosphatidylinositol synthase in the brain and retina of developing and adult rats. AB - CDP-diacylglycerol (CDP-DAG) synthase (CDS) is known as one of the key enzymes in the lipid synthesis including phosphoinositides (PIs) production. Phosphatidylinositol (PtdIns) synthase (PIS) catalyzes a formation of PtdIns from CDP-DAG in the PI cycle which produces several second messengers. We compared the gene expression for a presumably PI cycle specific-CDS molecule and a PIS using Northern blot analysis and in situ hybridization histochemistry in the central nervous system (CNS) and retina of developing and mature rats. Whereas no significant expression for CDS was detected during the prenatal stage in any CNS regions, PIS mRNA had already expressed on the prenatal day 15 throughout the neuroaxis including the spinal cord. During the postnatal stages, the gene expression for both CDS and PIS was detected widely in the gray matters throughout the entire brain. The expression for CDS was at higher levels in the olfactory mitral cells, the occipital cortex, the subiculum and hippocampal CA1 pyramidal cells, and the cerebellar Purkinje cells. On the other hand, the expression for PIS was at high levels in the olfactory mitral cells, the cerebral cortex, the hippocampal and dentate neuronal layer and the cerebellar Purkinje and granule cells. No significant expression for CDS or PIS was detected in the ventricular germinal zone, the cerebellar external granular layers, the mature ependyma or entire white matters. The expression for CDS and PIS decreased slightly throughout the CNS on P49. The significance of the parallel and discrepant expression patterns in terms of relative intensity between the two enzyme molecules was discussed in relation to the membrane turnover and signal transduction. PMID- 9733909 TI - Increase in phospholipase A2 activity towards lipopolymer-containing liposomes. AB - Phospholipase A2 (PLA2)-catalyzed hydrolysis of dipalmitoylphosphatidylcholine (DPPC) liposomes incorporated with submicellar concentrations of polyethyleneoxide covalently attached to dipalmitoylphosphatidylethanolamine (DPPE-PEG2000) has been studied in the gel-to-fluid transition region of the host DPPC lipid bilayer matrix. By means of fluorescence and light-scattering measurements, the characteristic PLA2 lag time has been determined as a function of lipopolymer concentration and temperature. The degree of lipid hydrolysis was followed using radioactive labeled lipids. Differential scanning calorimetry has been applied to characterize the thermodynamic phase behavior of the lipopolymer containing liposomes. A remarkable lipopolymer concentration-dependent decrease in the lag time was observed over broad temperature ranges. The radioactive measurements demonstrate an increase in catalytic activity for increasing amounts of lipopolymers in the bilayer. Hence, the lipopolymers act as a promoter of PLA2 lipid hydrolysis resulting in a degradation of the bilayer structure and a concomitant destabilization of the liposomes. This behavior is in contrast to the generally observed protective and stabilization effect in biological fluids exerted by lipopolymers in polymer-grafted liposomes. It is proposed that the enhanced activity of the small water soluble and interfacially active enzyme may involve a non-uniform distribution of the lipopolymers in the lipid matrix due to a coupling between local lipid bilayer curvature and composition of the non bilayer-preferring lipopolymers. PMID- 9733910 TI - Injections of D-amphetamine into the ventral pallidum increase locomotor activity and responding for conditioned reward: a comparison with injections into the nucleus accumbens. AB - The nucleus accumbens and ventral pallidum receive dopamine (DA) projections from the mesencephalon. Although DA inputs to the nucleus accumbens are implicated in both locomotion and reward processes, little is known of the behavioural significance of DA in the ventral pallidum. These studies examined the effects of D-amphetamine injected into the nucleus accumbens or ventral pallidum on locomotor activity and responding for a conditioned reward (CR). In the nucleus accumbens D-amphetamine dose dependently (1, 3 and 10 microg) increased locomotion within 5-10 min of injection. Intra-ventral pallidum microinjections of D-amphetamine also increased activity in this dose range, but the effect occurred with a longer latency (5-20 min). The magnitude of the response evoked by ventral pallidum injections was lower than that evoked by nucleus accumbens injections. The GABAA antagonist picrotoxin (0.1 microg) stimulated activity when injected into the ventral pallidum but not the nucleus accumbens, providing a pharmacological dissociation between the two injection sites. In the CR studies, D-amphetamine injected into both sites potentiated responding for a CR previously paired with food delivery, without altering responding on an inactive lever. Picrotoxin injected into the ventral pallidum reduced responding and abolished the selectivity of responding for CR. The results show that DA release in the ventral pallidum enhances locomotion and responding for a CR, providing evidence that DA in the ventral pallidum plays a significant role in the mediation of the effects of D-amphetamine. The failure of picrotoxin to elevate responding for CR despite increasing locomotor activity indicates that pharmacologically-induced blockade of GABAA receptors in the ventral pallidum disrupts goal-directed responding. PMID- 9733911 TI - Collagen type IV promotes the differentiation of neuronal progenitors and inhibits astroglial differentiation in cortical cell cultures. AB - In an effort to elucidate the interactions between cells in the developing cortex and their microenvironment, we have employed dissociated cell cultures and immunocytochemistry to analyze the effect of collagen type IV (COL) on the proliferation and differentiation of rat cortical progenitor cells during the period of corticogenesis. COL, present in the proliferative zones throughout the period of neurogenesis, belongs to a group of macromolecular proteins that make up a considerable portion of the extracellular matrix (ECM). We have shown that this ECM molecule inhibits cell proliferation and glial cell differentiation while promoting neuronal differentiation. We have also demonstrated that COL, when applied to the cultures with basic fibroblast growth factor (bFGF), induces glial cell differentiation while continuing to promote neuronal differentiation. These results indicate that cortical progenitor cells respond differentially to local environmental signals, and that components of the ECM are involved in the regulation of corticogenesis. PMID- 9733912 TI - Modification by ageing of the tetrodotoxin-sensitive sodium channels in rat skeletal muscle fibres. AB - Ageing leads to an impairment of muscle performance that may result from alteration of sarcolemma excitability. Therefore, we compare sodium channels of native fast-twitch skeletal muscle fibres of 21-26-month-old aged rats and 4-6 month-old young-adult rats, using the patch-clamp method. Extrajunctional sarcolemma of aged-rat fibres presented a higher sodium current density than that of young-rat fibres, which resulted from the presence of a higher number of available channels per membrane area. Open probability and availability voltage dependence of sodium channels were similar in aged- and young-rat fibres, but permeation property was altered during ageing: aged-rat muscles showed a bimodal distribution of fibres with two values of sodium-channel conductance measured between -40 and 0 mV; a young phenotype with a conductance close to 18 pS overlapping that found in young-rat fibres and an aged phenotype with a lower approximately half conductance. Current-voltage curves extended to -60 and +20 mV showed that the aged-phenotype conductance level resulted from an outward rectification occurring in these aged-rat fibres. Furthermore, in these aged-rat fibres belonging to the aged phenotype, ensemble average sodium currents showed slower activation and inactivation kinetics. Sodium currents of the two phenotypes were blocked by 100 nM tetrodotoxin, therefore excluding possible denervation effect. These age-related modifications in sodium current may contribute to the alteration of muscle excitability and function observed during the ageing process. PMID- 9733914 TI - Subgroups of hindbrain catecholamine neurons are selectively activated by 2-deoxy D-glucose induced metabolic challenge. AB - Glucose is a major fuel for body energy metabolism and an essential metabolic fuel for the brain. Consequently, glucose deficit (glucoprivation) elicits a variety of physiological and behavioral responses crucial for survival. Previous work indicates an important role for brain catecholamine neurons in mediation of responses to glucoprivation. This experiment was conducted to identify the specific catecholamine neurons that are activated by glucoprivation. Activation of hindbrain catecholamine neurons by the antimetabolic glucose analogue, 2-deoxy D-glucose (2DG; 50, 100, 200 or 400 mg/kg, s.c.) was evaluated using double label immunohistochemistry. Fos protein was used as the marker for neuronal activation and the enzymes tyrosine hydroxylase (TH) and phenethanolamine-N-methyl transferase (PNMT) were used as the markers for norepinephrine (NE) and epinephrine (E) neurons. 2-Deoxy-D-glucose (200 and 400 mg/kg) produced selective activation of distinct hindbrain catecholamine cell groups. In the ventrolateral medulla, doubly labeled neurons were concentrated in the area of A1/C1 and were predominantly adrenergic in phenotype. In the dorsal medulla, doubly labeled neurons were limited to C2 and C3 cell groups. In the pons, some A6 neurons were Fos-positive. Neurons in rostral C1, ventral C3, A2, A5 and A7 did not express Fos-ir in response to 2DG. Our results identify specific subpopulations of catecholamine neurons that are selectively activated by 2DG. Previously demonstrated connections of these subpopulations are consistent with their participation in the feeding and hyperglycemic response to glucoprivation. Finally, the predominant and seemingly preferential activation of epinephrine neurons suggests that they may play a unique role in the brain's response to glucose deficit. PMID- 9733913 TI - Cerebral metabolism following neonatal or adult hemineodecortication in cats: effect on oxidative capacity using cytochrome oxidase histochemistry. AB - In order to determine the degree and extent of changes in cerebral oxidative capacity following cerebral hemineodecortication, adult cats which had undergone surgery early postnatally (mean age: 11.4 days) or during adulthood were studied using cytochrome oxidase histochemistry. A total of 18 animals were employed and 50 brain regions were quantified bilaterally using optical densitometry. Although many subcortical regions exhibiting extensive degenerative features revealed lower levels of cytochrome oxidase (C.O.) activity, this reduction was relatively unremarkable compared to intact controls. Nevertheless, it was interesting that this decrease (down to 66-89%) of normal was more pronounced in neonatal-lesioned cats, reaching significance in a number of ipsilateral thalamic nuclei, compared to adult-lesioned animals (91-100% of normal), suggesting a contribution of glial cells to the density of C.O. staining in the latter cats. Regions of the brain spared from degeneration exhibited a bilateral increase in C.O. activity which may reflect the demands for energy to support the anatomical reorganization which is prevalent in these animals. Surprisingly, such increases were more robust within spared regions of the adult-lesioned brain, reaching significance in four ipsilateral and nine contralateral areas with the density of the reaction attaining levels over 125% of control. This may indicate different demands for oxidative metabolism in the adult-lesioned cats. These results enhance our understanding of the mechanism(s) underlying the greater extent of functional sparing or recovery in cats sustaining injury to the cerebral cortex early vs. late in life. In addition, the findings complement our previous companion report on glucose metabolism supporting the concept of energy compartmentalization, which reflects the dynamic interaction between anatomical and functional changes in this age-at-lesion model of recovery. PMID- 9733915 TI - A novel bacteriocin with a YGNGV motif from vegetable-associated Enterococcus mundtii: full characterization and interaction with target organisms. AB - A novel broad-spectrum antimicrobial peptide produced by vegetable-associated Enterococcus mundtii was purified and characterized, and designated mundticin. To our knowledge, this is the first report on bacteriocin production by this organism. The elucidation of the full primary amino acid sequence of mundticin (KYYGNGVSCNKKGCSVDWGKAIGIIGNNSAANLATGGAAGWSK) revealed that this antimicrobial peptide belongs to the class IIa bacteriocins of lactic acid bacteria which share a highly conserved N-terminal 'YGNGV' motif. Data obtained by computer modelling indicated an oblique orientation of the alpha-helical regions of mundticin and homologous class IIa bacteriocins at a hydrophobic-hydrophilic interface, which may play a role in the destabilization of phospholipid bilayers. The average mass of mundticin, as determined by electron spray mass spectrometry, was found to be 4287.21+/-0.59 Da. With respect to its biological activity, mundticin was shown to inhibit the growth of Listeria monocytogenes, Clostridium botulinum and a variety of lactic acid bacteria. Moreover, it was demonstrated to have a bactericidal effect on L. monocytogenes as a result of the dissipation of the membrane potential, and a loss of intracellular ATP in absence of ATP leakage. Its good solubility in water, and its stability over a wide pH and temperature range indicate the potential of this broad spectrum bacteriocin as a natural preservation agent for foods. PMID- 9733916 TI - A comparison of the adenosine-mediated synaptic inhibition in the CA3 area of immature and adult rat hippocampus. AB - We compared the effects of the adenosine A1 receptor activation on the postsynaptic potentials (psps) recorded from the CA3 area of immature (postnatal days 10-20) and adult rat hippocampal neurons in vitro. The adenosine A1 receptor agonist 2-phenyl-isopropyl-adenosine (PIA, 1 microM) depressed the stimulus induced psps less in immature and more in adult neurons. In the presence of the GABAA receptor antagonist bicuculline methiodide (BMI, 10 microM), PIA reduced the duration and number of action potentials of the stimulus-induced paroxysmal depolarizations (PDs) in immature neurons, while it blocked PDs in adult neurons. Spontaneous BMI-induced PDs, were blocked by PIA in less than half (5/12) immature and all (6/6) adult neurons. The adenosine A1 receptor antagonist 8 cyclopentyl-1,3-dipropylxanthine (DPCPX, 1 microM) enhanced the stimulus-induced psps in immature and adult neurons alike; this effect did not lead to stimulus induced bursting in immature neurons. DPCPX induced spontaneous bursts (proconvulsant effect) in only 2/16 immature but in all adult (12/12) neurons. In BMI, DPCPX increased the duration and number of action potentials of the stimulus induced PDs in immature and adult neurons alike (by about 30%), but it increased the rates of occurrence of spontaneous PDs in significantly more adult neurons. In conclusion, our results suggest that adenosine, acting via A1 receptors, is a more effective endogenous anti-epileptic in adult than in immature hippocampus, a fact which may contribute to the susceptibility of the latter to epileptogenesis. PMID- 9733917 TI - Genetic analysis of behavioral, neuroendocrine, and biochemical parameters in inbred rodents: initial studies in Lewis and Fischer 344 rats and in A/J and C57BL/6J mice. AB - Previous work has identified inherent behavioral, neuroendocrine, and biochemical differences among inbred rodent strains that have been related to the animals' differential responsiveness to drugs of abuse or stress. In the present study, we sought to determine (1) whether there are genetic correlations among particular phenotypic traits that differ between a pair of inbred rat strains (Lewis and Fischer 344) or a pair of inbred mouse strains (A/J and C57BL/6J); (2) which of these traits might be amenable to quantitative trait locus analysis; and (3) whether additional behavioral or biochemical differences relevant to drug- or stress-responsiveness could be identified in these strains. Specifically, we measured several behavioral, neuroendocrine, and biochemical traits in parental Lewis and Fischer 344 rats and in 298 members of an F2 intercross population, as well as in parental A/J and C57BL/6J mice and in 11 of the AXB/BXA recombinant inbred mouse strains. Traits measured included exploratory locomotor activity in a novel environment; amphetamine-induced locomotor activity; several specific protein levels in striatal regions, including inhibitory G protein subunits, the dopamine transporter, the Fos family member transcription factor DeltaFosB, and the protein phosphatase inhibitor DARPP-32; and late-afternoon plasma corticosterone concentrations. Each of the traits measured in F2 rats or recombinant inbred mice appears to be influenced by multiple genes, as well as by environmental factors. There were statistically significant, albeit relatively weak, correlations among several traits in an F2 intercross population bred from Lewis and Fischer rats. Among the traits studied in Lewis and Fischer rats, one seemed most amenable to quantitative trait locus analysis: the level of the inhibitory G-protein subunit, Galphai, in the nucleus accumbens. We also found a robust genetic correlation between levels of DeltaFosB and levels of the dopamine transporter in striatal regions in AXB/BXA recombinant inbred mouse strains. While these studies demonstrate the likely complexity of the genetic factors that influence the numerous phenotypes associated with altered responsiveness to drugs of abuse and stress, they represent an initial and necessary step toward identifying specific genetic factors involved. PMID- 9733918 TI - Cytogenetic effects of copper-containing intrauterine contraceptive device (IUCD) on blood lymphocytes. AB - Forty women using the copper-containing intrauterine contraceptive device (IUCD) for various periods and 22 control women were studied. The copper and zinc levels were measured in the plasma of the IUCD users and the control group. Chromosomal analyses were performed on blood lymphocytes of the same groups. Results showed that plasma copper level was significantly higher in the IUCD users than in that of controls (1.25+/-0.14 microg/ml vs. 0.89+/-0.04 microg/ml, P<0. 05). A decline in plasma zinc level from 2.90+/-0.41 microg/ml to 1. 27+/-0.13 microg/ml, P<0.05, with a significant alteration in plasma copper/zinc ratio was also measured in the IUCD women (62% vs. 111%, P<0.05). Chromosomal aberrations (4.47+/-1.40 vs. 0.62+/-0.12/100 cell, P<0.01) and high frequencies of sister chromatid exchanges (8. 79+/-0.46 vs. 6.59+/-0.19 SCE/cell, P<0.01) were evident in the IUCD women especially in those using the IUCD for more than 24 months. The combination of high copper plasma level, chromosomal aberrations and the increased frequency of sister chromatid exchange may support the existence of a positive correlation between the long term use of the IUCDs and DNA damage in the host somatic cells. PMID- 9733919 TI - Out-to-in translocation of butanetriol-containing phospholipid analogs in human erythrocyte membrane. AB - Fluorescent butanetriol-containing phospholipid analogs were synthesized by replacing the glycerol moiety in 1-hexadecanoyl-2-[6-N-(7-nitrobenz-2-oxa-1,3 diazol-4-yl) aminohexanoyl]-sn-glycero-3-phosphocholine, -phosphoethanolamine, phosphoserine and 1-hexadecanoyl-2-[12-N-(7-nitrobenz-2-oxa-1, 3-diazol-4 yl)aminododecanoyl]-sn-glycero-3-phosphocholine, -phosphoethanolamine, phosphoserine by the 1,3,4-butanetriol residue, and their out-to-in translocation in the human erythrocyte membrane studied by 'back exchanging' the outer surface incorporated phospholipids using bovine serum albumin. The results of these studies indicate that the replacement of the glycerol moiety by the 1,3,4 butanetriol residue in aminophospholipids does not effect their out-to-in translocation in the human erythrocyte membrane. Furthermore, since earlier study by Arora and Gupta (Biochim. Biophys. Acta 1324 (1997) 47-60) has shown that the conformation of the 1,3,4-butanetriol phospholipids possess the backbone conformation similar to that of glycerophospholipids, it is suggested that besides the normal phospholipid polar head-group, a normal phospholipid interface conformation may also be required for the aminophospholipid-translocase interactions. PMID- 9733920 TI - In contrast to cocaine, prenatal exposure to methadone does not produce detectable alterations in the developing mouse brain. AB - Whereas prenatal cocaine exposure dramatically alters brain development, the safety of methadone in detoxification programs for heroin-addicted pregnant women is uncertain. This paper compares the effects of exposure to methadone or to cocaine in utero on a model system, the developing mouse brain. Methadone (40 mg/kg/day, i.e., 40-fold detoxification dosage) or cocaine (30 mg/kg/day, as in severe addiction) was injected into mice from day 8 to day 18 of gestation. Pre- and postnatal brain development was analyzed at the anatomical and microscopical levels, including by immunostaining of post-mitotic cells, neurites, and astrocytes. Prenatal mice exposure to cocaine caused neuronal misaddressing among neocortical layers, abnormal gliogenesis, and defective neuritic outgrowth and bundling. Methadone produced small-for-date offspring with normal brain development. In conclusion, supratherapeutic methadone doses induce intrauterine growth retardation in mice, but spare brain cytoarchitecture. In contrast, cocaine produces less growth retardation, but severely disturbs neocortical layering. PMID- 9733921 TI - Molecular analysis of mutations induced by acrolein in human fibroblast cells using supF shuttle vector plasmids. AB - Types of mutations induced by acrolein in the supF gene on the shuttle vector plasmid pMY189 replicated in normal human fibroblast cells were examined. Base sequence analysis of 92 plasmids with mutations in the supF gene revealed that the majority of the mutations were base substitutions (76%) and the others were deletions and insertions (24%). Single base substitutions were most frequently found (46%), while multiple base substitutions were 18% and tandem (two adjacent) base substitutions were 12% of the mutations. Of the base substitution mutations, G:C to T:A transversions were 44% and G:C to A:T transitions were 24%. The mutations were distributed not randomly but located at several hotspots. Acrolein produced DNA intra-strand cross-links between guanine residues, which might be responsible for rather high induction of the tandem base substitution mutations. PMID- 9733922 TI - Properties of lipoamino acids incorporated into membrane bilayers. AB - Several lipoamino acids were synthesized in which palmitic acid was coupled with the alpha-amino group of an amino acid. These lipoamino acids were tested for their inhibitory action against Sendai virus fusion to liposomes composed of egg phosphatidylethanolamine and 5 mol% of the ganglioside GD1a. A commonly employed viral fusion assay based on the dilution of the fluorescent probe octadecylrhodamine (R18) exhibited an additional complication in the presence of Nalpha-palmitoyl tryptophan (palm-Trp). At higher mol fraction of palm-Trp it was observed that there was an increase in R18 quenching. Studies on the dependence of the emission wavelength of palm-Trp on excitation wavelength demonstrated that the presence of R18 alters the environment of the indole. The results illustrate one of the complexities of viral fusion assays using the R18 probe. Despite this complication it was possible to demonstrate that several of the lipoamino acids are effective at inhibiting the fusion of Sendai virus to liposomes as measured by the R18 assay. One of the most effective inhibitors of this process is palm Trp which, at a concentration of 4 mol% in liposomes, markedly reduces the apparent rate of fusion. At pH 5.0 this amphiphile is also an inhibitor of Sendai virus fusion, indicating that the ionization of the carboxyl group of this amphiphile is not required for its antiviral activity. The inhibitory action of palm-Trp against Sendai virus was confirmed by demonstrating inhibition of Sendai mediated cytopathic effects studied in tissue culture. A property associated with antiviral activity is the ability of amphiphiles to raise the bilayer to hexagonal phase transition temperature of dielaidoyl phosphatidylethanolamine. All of these lipoamino acids were found to possess this property, but a quantitative relationship with inhibition of viral fusion was not found. PMID- 9733923 TI - Synaptic effects of nitric oxide on enkephalinergic, GABAergic, and glutamatergic networks of the rat periaqueductal gray. AB - Previous studies have shown that the injection of nitric oxide (NO) donating compounds into the dorsal periaqueductal gray region of the midbrain (PAG) decreases mean arterial pressure (MAP), while the injection of NO synthase (NOS) inhibitors increases MAP. In this study we used both in-vivo and in-vitro preparations and examined the effect of a NO donor and a NOS inhibitor on MAP, membrane properties, and synaptic activities in PAG neurons. We found that: (1) Injection of the NO donor hydroxylamine (HA) into the dorsal PAG decreased MAP, while the injection of the neuronal NOS (nNOS) inhibitor, 1-(2 trifluoromethylphenyl) imidazole (TRIM) increased MAP. These responses were consistent and site-specific. (2) HA-evoked hypotensive responses were mediated by PAG neuronal activity, because they were blocked by pre-injection with gamma amino-butyric acid (GABA). (3) HA consistently increased the rate of observable synaptic events while TRIM consistently decreased the rate of observable synaptic events. (4) Bicuculline (BIC) and naloxone (NAL) blocked HA-evoked increases in the rate of observable inhibitory synaptic events. (5) Perfusion with sodium nitroprusside (SNP) and illumination with bright light consistently elevated rates of observable synaptic events, and SNP-evoked increases of excitatory synaptic events were blocked by pretreatment with glutamic acid antagonists. (6) PAG-medullary projecting neurons exhibited similar response patterns. The results of this study suggest that: (1) NO production within the PAG is a major component of PAG-mediated cardiovascular responses. (2) The effects of NO may be mediated in part by increased presynaptic vesicular release of glutamic acid, GABA, and enkephalin. PMID- 9733924 TI - Expression of olfactory receptors, G-proteins and AxCAMs during the development and maturation of olfactory sensory neurons in the mouse. AB - Three mouse olfactory receptors have been cloned and sequenced and were found to be expressed in different zones of the olfactory epithelium. In situ hybridisation (ISH) results showed that each olfactory receptor was expressed at an early stage in development (E12), was not dependent on the maturation of the receptor neurons, and was present long before the onset of odour detection. Cells positive for these same olfactory receptors and the G-protein (Gbeta) were also found in non-neural regions of the nasal epithelium in the earlier stages of development (E12-16). Ncam, and Big-2 expression were, however, restricted to the region of developing olfactory neurons. Ncam expression appeared in advance of the olfactory receptor expression, while Big-2 appeared after olfactory receptor expression and neither were expressed in cells outside the olfactory epithelium. Both showed the highest number of positive cells in the early post-partum period when olfactory detection is functional. Ncam is known to be involved in guidance of the developing olfactory axons and was expressed earlier than any of the olfactory receptors, while Big-2 appears somewhat later (E14) at a time when developing axons reach the olfactory bulb. Moreover the highest periods of expression occur at post-natal day 7 when a proliferation of bulbar glomeruli are observed, suggesting the role of Big-2 to be primarily concerned with synaptogenesis. PMID- 9733925 TI - Modulatory effects of melatonin on genotoxic response of reference mutagens in the Ames test and the comet assay. AB - The effect of a potent endogenous antioxidant, the pineal gland indole melatonin (MLT) on the mutagenicity of twelve well-known mutagens and carcinogens has been investigated using two in vitro tests the Ames test and the single cell gel electrophoresis assay (SCGE assay or COMET assay). The 12 mutagens used were 7, 12-dimethylbenz(a)anthracene (DMBA), benzo(a)pyrene (BP), 2-aminofluorene (AF), 1,2-dimethylhydrazine (DMH), bleomycin, cyclophosphamide (CP), 4-nitroquinoline-N oxide (NQO), 2,4, 7-trinitro-9-fluorenone (TNF), 9-aminoacridine (AA), N nitrosomethylurea (NMU), mitomycin C and sodium azide tested in the absence or in the presence of S9 mix. MLT alone turned out neither toxic nor mutagenic in the Ames test and revealed clastogenic activity at the highest concentration tested (100 microM) in the SCGE assay. In four Salmonella typhimurium tester strains TA 97, TA 98, TA 100 and TA 102 MLT significantly reduced the mutagenicity of chemicals which require S9 activation. In the SCGE assay performed on CHO cells, preincubation with MLT led to a strong inhibition of clastogenic activities of DMBA and CP, and in a lesser extent with BP and NMU. With mitomycin C, MLT exacerbated responses in both tests. The possible mechanisms of MLT's inhibitory action are discussed. PMID- 9733926 TI - The structuring effects of amphotericin B on pure and ergosterol- or cholesterol containing dipalmitoylphosphatidylcholine bilayers: a differential scanning calorimetry study. AB - Amphotericin B (AmB) is the most widely used polyene antibiotic to treat systemic fungal infections which affect an increasing number of immunocompromised patients. It is generally thought that AmB forms pores within the fungi membranes by interacting with ergosterol, the main sterol of fungi. However, it also interacts with the cholesterol contained in mammalian cells, hence its toxicity. In order to have a better understanding of the interactions prevailing between AmB and sterols, differential scanning calorimetry was used to study various mixtures incorporating from 6.5 to 25 mol% of AmB in pure dipalmitoylphosphatidylcholine (DPPC) vesicles and in ergosterol- or cholesterol containing DPPC vesicles. The sterol concentration was kept constant at 12.5 mol% with respect to the phospholipid. Our results show that three phases co-exist when AmB is dispersed in the pure phospholipid. One corresponds to the phospholipid phase alone. The two others are characterised by a broad transition at temperatures higher than the main transition temperature of the pure phospholipid, corresponding to the drug in interaction with the aliphatic chains of the lipid. The fact that the transition temperatures of these additional components are higher than that of the pure phospholipid suggests that AmB interacts strongly with the aliphatic chains of the lipid, consistent with the idea prevailing in the literature that AmB by itself may form pores in a lipid matrix. When AmB interacts with cholesterol-containing bilayers the thermograms also present three components. Upon increasing the concentration of AmB, though, an important broadening of these components is observed which is explained in terms of destabilisation of the organisation of the aliphatic chains. The situation is strikingly different if ergosterol is present in the lipid matrix. The thermograms remain unmodified as the concentration of AmB is increased and a broad transition, now involving only two components when the thermograms are decomposed, is observed. An analysis of the results shows that various interacting units, e.g. AmB+DPPC and (AmB+ergosterol)+DPPC, are present within the membrane. These units involve the phospholipid and hence contribute to its structurisation. The important differences between the thermograms obtained with the ergosterol- as compared to the cholesterol-containing bilayers, in spite of the structural similarity of these two sterols, provides strong evidence for the selectivity of interaction of AmB with ergosterol as compared to cholesterol. It is thus clear that the action of AmB on cholesterol- as compared to ergosterol containing membranes results from different mechanisms. Finally, UV-visible spectra of AmB in pure as well as sterol-containing DPPC vesicles show the presence of absorption bands that give support to the interpretation derived from the calorimetric data. PMID- 9733928 TI - Modifying effect of vitamins C, E and beta-carotene against gamma-ray-induced DNA damage in mouse cells. AB - The modifying effect of treatment with vitamins C, E and beta-carotene on the clastogenic activity of gamma rays was investigated in mice. Damage in vivo was measured by the micronucleus assay in bone marrow polychromatic erythrocytes and exfoliated bladder cells. The vitamins were administered orally, either for five consecutive days before or immediately after irradiation with 2 Gy of gamma rays. The results show that pretreatment with vitamin E (100-200 mg/kg/day) and beta carotene (3-12 mg/kg/day) were effective in protecting against micronucleus induction by gamma rays. Vitamin C depending on its concentration enhanced the radiation effect (400 mg/kg/day), or reduced the number of micronucleated polychromatic erythrocytes (50-100 mg/kg/day). Such effect was weekly observed in exfoliated bladder cells. The most effective protection in both tissues was noted when a mixture of these vitamins was used as a pretreatment. Administration of the all antioxidant vitamins to mice immediately after irradiation was also effective in reducing the radiation-induced micronucleus frequency. The data from the in vitro experiments based on the comet assay show that the presence of the vitamins in culture medium influences the kinetic of repair of radiation-induced DNA damage in mouse leukocytes. PMID- 9733927 TI - Postnatal expression pattern of calcium-binding proteins in organotypic thalamic cultures and in the dorsal thalamus in vivo. AB - The present study describes the postnatal expression of calbindin, calretinin and parvalbumin and glutamic acid decarboxylase (GAD) and microtubule-associated protein 2 (MAP2) in organotypic monocultures of rat dorsal thalamus compared to the thalamus in vivo. Cultures were maintained for up to 7 weeks. Cortex conditioned medium improved the survival of thalamic cultures. MAP2 immunoreactive material was present in somata and dendrites of small and large sized neurons throughout the cultures. Parvalbumin immunoreactivity was present in larger multipolar or bitufted neurons along the edge of a culture. These neurons also displayed strong parvalbumin mRNA and GAD mRNA expression, and GABA immunoreactivity. They likely corresponded to cells of the nucleus reticularis thalami. Parvalbumin mRNA, but neither parvalbumin protein nor GAD mRNA, was expressed in neurons with large somata within the explant. They likely represented relay cells. GAD mRNA, but not parvalbumin mRNA, was expressed in small neurons within the explants. Small neurons also displayed calbindin- and calretinin-immunoreactivity. The small neurons likely represented local circuit neurons. The time course of expression of the calcium-binding proteins revealed that all were present at birth with the predicted molecular weights. A low, but constant parvalbumin expression was observed in vitro without the developmental increase seen in vivo, which most likely represented parvalbumin from afferent sources. In contrast, the explantation transiently downregulated the calretinin and calbindin expression, but the neurons recovered the expression after 14 and 21 days, respectively. In conclusion, thalamic monocultures older than three weeks represent a stable neuronal network containing well differentiated neurons of the nucleus reticularis thalami, relay cells and local circuit neurons. PMID- 9733929 TI - Modulation of the vacuolar H+-ATPase by adenylates as basis for the transient CO2 dependent acidification of the leaf vacuole upon illumination. AB - Using tonoplast vesicles, we have investigated the activity of the vacuolar H+ ATPase which is the dominant proton pump at the tonoplast of mesophyll cells. Bafilomycin-sensitive ATP hydrolysis or acidification of tonoplast vesicles in the presence of ATP were measured at varying ATP, ADP and Pi concentrations, and in the presence of oxidized or reduced glutathione. Increased ATP/ADP ratios as reported for the extrachloroplast cytoplasm during the induction phase of photosynthesis at high or low CO2 (P. Gardestrom, Biochim. Biophys. Acta 1183 (1993) 327-332) increased the activity of the V-ATPase in simulation experiments with vesicles. Depending on reported subsequent decreases in cytoplasmic ATP/ADP ratios in the presence of high or low CO2, the ATPase activity of tonoplast vesicles changed in simulation experiments to lower values. More than 10 mM phosphate was required to decrease the ATPase activity in vesicles significantly at ATP/ADP ratios of 3 or higher, indicating that ATPase activity is controlled more by ratios of ATP to ADP than by phosphorylation potentials (ATP)/(ADP)(Pi). Oxidized glutathione was inhibitory. The results permit interpretation of the observation that on illumination of previously darkened leaves the pH of the vacuoles of mesophyll cells decreases indicating energized transport of protons across the tonoplast into acidic vacuoles, and that the extent of vacuolar acidification depends on the CO2 concentration of the surrounding air (Z.-H. Yin, S. Neimanis, U. Heber, Planta 182 (1990) 253-261). We conclude that short term control of tonoplast ATPase activity in leaves during dark/light transients can essentially be understood on the basis of reported changes in cytoplasmic ATP/ADP ratios, with a possible participation of redox modulation. PMID- 9733930 TI - Chronic ethanol administration alters the modulatory effect of 5alpha-pregnan 3alpha-ol-20-one on the binding characteristics of various radioligands of GABAA receptors. AB - In this study, we investigated the modulatory effect of 5alpha-pregnan-3alpha-ol 20-one, a neurosteroid, on the binding characteristics of [3H]flunitrazepam (2 nM), [3H]muscimol (5 nM), and 4 nM [35S]t-butylbicyclophosphorothionate (TBPS) in cerebral cortex, cerebellum, and hippocampus of control, ethanol-dependent, and ethanol-withdrawn rats. 5alpha-Pregnan-3alpha-ol-20-one potentiated the binding of [3H]flunitrazepam and [3H]muscimol in all the rat brain regions investigated in this study. There was a significant increase in the maximal potentiation of [3H]flunitrazepam as well as [3H]muscimol binding (Emax) in the ethanol-dependent rat cerebellum as compared to control group (p<0. 025). Furthermore, 5alpha pregnan-3alpha-ol-20-one elicited a biphasic response, i.e., it potentiated the binding of [35S]TBPS at lower concentrations (<=100 nM) and inhibited the binding at higher concentrations (>100 nM). There was a significant higher inhibition of [35S]TBPS binding (-Emax) by 5alpha-pregnan-3alpha-ol-20-one in the hippocampus of ethanol-dependent as well as ethanol-withdrawn rats (p<0.025). These observations suggest that the neurosteroid binding site associated with the gamma aminobutyric acidA (GABAA) receptors in cerebellum and hippocampus plays an important role during ethanol-dependence and ethanol-withdrawal, and some of the changes following ethanol dependence and its withdrawal may be mediated through the neurosteroid binding site. PMID- 9733931 TI - Effects of liposome-encapsulated drugs on macrophages: comparative activity of the diamidine 4',6-diamidino-2-phenylindole and the phenanthridinium salts ethidium bromide and propidium iodide. AB - Liposomes can be used for the intracellular delivery of drugs into macrophages. Previously, we developed a liposome-mediated macrophage 'suicide' technique based on the intraphagocytic accumulation of the liposomally delivered bisphosphonate clodronate. Later we found that the diamidine propamidine is even more effective in this approach. In the present study it is shown that liposome-encapsulated 4', 6-diamidino-2-phenylindole (L-DAPI), another well known DNA-binding diamidine, is the most effective drug in killing liver macrophages (Kupffer cells), when intravenously administered in rat. Compared to liposome-encapsulated propamidine (L-propamidine) it showed about 10-fold more activity on a molar basis. Furthermore, L-DAPI was found to induce cell death by inducing apoptosis. The structurally strongly related phenanthridinium salts ethidium bromide (EB) and propidium iodide (PI) exert marked differences in their efficacy. Whereas liposome-encapsulated PI (L-PI) was about 5 times more active in killing macrophages than L-propamidine, liposome-encapsulated EB (L-EB) showed a strongly reduced activity (10 times less than L-PI). As is shown here, PI remains mainly encapsulated in liposomes, while substantial amounts of EB leak out of liposomes. This may very well explain the differences in in vivo activity between L-EB and L PI. PMID- 9733933 TI - Genotoxicity of Farbasol and its component: 4-ethyltoluene. AB - A combination of assays for gene mutations in Salmonella typhimurium TA97a, TA98, TA100 and TA102 strains with and without rat liver activation, and for micronucleus and sister chromatid exchange (SCE) in bone marrow cells of Imp:Balb/c mice was used to provide data on the mutagenic and genotoxic properties of the mixture of aromatic solvents, known under the trade name of Farbasol. In addition, 4-ethyltoluene (the main ethylmethylbenzenic component of Farbasol) was also tested for muta- and genotoxicity. The results revealed that neither Farbasol nor 4-ethyltoluene induced an increased reverse mutation in bacterial cells or the formation of micronucleated polychromatic erythrocytes in bone marrow. However, those compounds were found to be active as sister chromatid exchange (SCE) agents. PMID- 9733932 TI - Lipopolysaccharide-induced fever is dissociated from apoptotic cell death in the rat brain. AB - Immune system activation induces increase in expression level and enzymatic activity of interleukin-1 beta converting enzyme (ICE) in rat brain. As ICE has been implicated in apoptotic cell death, a possible link may exist between immune system activation by bacterial endotoxic lipopolysaccharide (LPS) and apoptosis in rat brain. The aim of this study was to investigate possible effect of acute (5.5 h) or chronic (5 days) intraperitoneal (i.p.) administration and central injection of LPS on brain apoptotic cell death. Body temperature was continuously monitored for fever, a hallmark of immune activation. Detection of apoptotic cell death was carried out by using in situ labelling of DNA fragmentation in various brain structures. Despite the chronic or the acute pyrogenic effects of LPS, no evidence for apoptotic cell death was observed in any of the brain areas analysed, including hippocampus, hypothalamus, area postrema, subfornical organ, organum vasculosum of the lamina terminalis and nucleus tractus solitaris. Other well-known sites of apoptotic cell death, including brain of ischemic rat, mammary gland of post-lactating rat and rat intestine as well as Dnase-treated rat brain slices, were used as positive controls. These results suggest that ICE activation during fever development is dissociated from cell death by apoptosis in rat brain. Unlike peripheral targets of immunocompetent cytokines, a protective system, yet to be defined, may be present in the central nervous system and block the deleterious effects of infectious agents and cytokines. PMID- 9733934 TI - Regulation of extracellular potassium in the developing hippocampus. AB - It has long been felt that one of the reasons that the immature brain has an increased propensity to seize is due to an inability to regulate the extracellular potassium ([K+]o). However, the data supporting this hypothesis is controversial. Here, we tested the regulation of [K+]o in urethane-anesthetized juvenile and adult rats during and after synchronized epileptiform activity induced by 20 Hz stimulus trains to the CA3 region. The regulation of [K+]o in CA3 was compared to the same measurements in CA1. Across all age groups, there was no difference between CA1 and CA3. There was a slight decrease in the peak level of [K+]o reached during stimulation only in the youngest age groups tested (PN9-11). There was an age-dependent change in the rate of recovery of [K+]o from the elevated levels. This recovery was slowest in the PN9-11 age group. In all the animals in the PN9-11 group and 3/7 animals in the PN14-15 group, there was a secondary increase in the [K+]o during the afterdischarge. This secondary rise was never observed in animals over 15 days of age. These data confirm that there is altered regulation of [K+]o in the developing brain that takes two forms: (1) the ability to recover from elevated [K+]o levels, suggesting maturation of regulatory, or uptake, mechanisms; and (2) that which is related to mechanisms of synchronization and initiation of afterdischarges and the level of [K+]o that is produced during the discharge. PMID- 9733935 TI - Surface plasmon resonance analysis at a supported lipid monolayer. AB - Methods for the formation of supported lipid monolayers on top of a hydrophobic self assembled monolayer in a surface plasmon resonance instrument are described. Small unilamellar vesicles absorb spontaneously to the surface of the hydrophobic self-assembled monolayer to form a surface which resembles the surface of a cellular membrane. Lipophilic ligands, such as small acylated peptides or glycosylphosphatidylinositol-anchored proteins, were inserted into the absorbed lipid and binding of analytes to these ligands was analysed by surface plasmon resonance. Conditions for the formation of lipid monolayers have been optimised with respect to lipid type, chemical and buffer compatibility, ligand stability and reproducibility. PMID- 9733936 TI - Micronuclei in lymphocytes and exfoliated buccal cells of postmenopausal women with dietary changes in folate. AB - Folate deficiency is associated with anemia, birth defects, cancer and neuropsychiatric disorders. The purpose of this study was to determine if a moderate folate deficiency during controlled changes in folate intake would affect chromosomal damage in lymphocytes and buccal cells. A study of nine healthy postmenopausal women volunteers (age 49-63 years) was carried out in a metabolic unit (baseline week with folate intake of 195 microg/day, five-week depletion at 56 microg/day, and gradual repletion including four weeks at 111 microg/day, 11 days at 286 microg/day and 9 days at 516 microg/day). Plasma folate, vitamin B-12, and homocysteine were measured weekly. Cytogenetic damage was assessed by scoring micronucleus (MN) frequency in lymphocytes and buccal cells three times: (1) at the beginning of the study, (2) at the end of depletion, and (3) after repletion. The MN frequency increased in binucleated lymphocytes, as well as in all lymphocytes, after depletion (p=0.037), and later decreased following repletion (p=0. 028). Both kinetochore-positive and kinetochore-negative MN were increased after depletion (p=0.015 and 0.028), but after repletion only the change in kinetochore-positive MN was statistically significant (p=0.048). The main variables affecting MN were: (1) vitamin B-12 level, (2) plasma folate level, and (3) baseline frequency of MN. The MN frequency in exfoliated buccal cells was decreased after dietary supplementation of 516 microg/day folate (p=0.010). Thus, low folate, without clinical symptoms of anemia, results in higher levels of cytogenetic damage in both the blood and oral cavity of postmenopausal women. PMID- 9733937 TI - Developmental modulation of mouse hypoglossal nerve inspiratory output in vitro by noradrenergic receptor agonists. AB - The ontogeny of the noradrenergic receptor subtypes modulating hypoglossal (XII) nerve inspiratory output was characterized. Noradrenergic agents were locally applied over the XII nucleus of rhythmically active medullary slice preparations isolated from mice between zero and 13 days of age (P0-P13) and the effects on XII inspiratory burst amplitude quantified. The alpha1 receptor agonist phenylephrine (PE, 0.1-10 microM) produced a dose-dependent, prazosin-sensitive (0.1-10 microM) increase in XII nerve inspiratory burst amplitude. The magnitude of this potentiation increased steadily from a maximum of 15+/-8% in P0 mice to 134+/-4% in P12-P13 mice. The beta receptor agonist isoproterenol (0.01-1.0 mM) produced a prazosin-insensitive, propranolol-sensitive potentiation of XII nerve burst amplitude. The isoproterenol-mediated potentiation increased with development from 27+/-5% in P0-P1 slices, to 37+/-3% in P3 slices and 45+/-4% in P9-P10 slices. The alpha2 receptor agonist clonidine (1 mM) reduced XII nerve inspiratory burst amplitude in P0-P3 slices by 29+/-5%, but had no effect on output from P12-P13 slices. An alpha2 receptor-mediated inhibition of inspiratory activity in neonates (P0-P3) was further supported by a 19+/-3% reduction in XII nerve burst amplitude when norepinephrine (NE, 100 microM) was applied in the presence of prazosin (10 microM) and propranolol (100 microM). Results indicate that developmental increases in potentiating alpha1 and, to a lesser extent, beta receptor mechanisms combine with a developmentally decreasing inhibitory mechanism, most likely mediated by alpha2 receptors, to determine the ontogenetic time course by which NE modulates XII MN inspiratory activity. PMID- 9733938 TI - Ontogeny of the GNRH-, glutaminase- and glutamate decarboxylase-gene expression in the hypothalamus of female rats. AB - Amino acid neurotransmitters like gamma-aminobutyric acid (GABA) and glutamate (GLU) are involved in the regulation of hypothalamic gonadotropin releasing hormone (GnRH) release. We investigated, whether there are changes of gene expression in the rat hypothalamus for GnRH, GnRH receptor, as well as glutaminase and glutamate decarboxylase, two enzymes regulating neurotransmitter concentrations of GLU and GABA in the brain during the ontogeny. After reverse transcription-polymerase chain reaction (RT-PCR) we used an ELISA method to quantify PCR products. In 15-day old animals high plasma luteinizing hormone (LH) levels with pronounced variations were found. In 25-day old animals LH values were low, whereas in 35-day old rats LH levels increased significantly indicating the reactivation of the GnRH-pulse generator at the beginning of puberty. In parallel to these changes, the mRNA levels of the GnRH receptor in the mediobasal hypothalamus were high at day 15, significantly lower at day 25 and again high at day 35 after birth (ELISA O.D. GnRH-R day 15: 0.46+/-0.07, day 25: 0.16+/-0.04, day 35: 0.36+/-0.04; p<0.01), but no changes of GnRH receptor gene expression were found in the preoptic area. The mRNA of GnRH in the preoptic area as well as mRNA levels of glutaminase and glutamate decarboxylase in the mediobasal hypothalamus and the preoptic area did not change during ontogeny. We conclude that hypothalamic GnRH receptors are involved in the characteristic changes of LH secretion patterns during sexual maturation. Major changes of GnRH receptor gene expression occurred in the mediobasal hypothalamus and correlated well with plasma LH levels, whereas hypothalamic mRNA levels of GnRH, glutaminase and glutamate decarboxylase did not change within the different age groups. Thus the activity of the GABA- and glutamatergic system during ontogeny may be regulated at the receptor or postreceptor level. PMID- 9733939 TI - Detergent solubilisation of phospholipid bilayers in the gel state: the role of polar and hydrophobic forces. AB - Testing the solubilisation of phosphatidylcholine (PC) bilayers by Triton X-100 reveals that in the gel state, but not in the fluid state, the amount of detergent required to solubilise the phospholipid is highly dependent on the chain length. Saturated C16 and C18 PC are virtually insoluble at 4 degreesC. However, addition of water-soluble reagents that perturb hydrogen bonding, e.g. urea, or of small proportions of non-bilayer lipids, make the bilayers amenable to detergent solubilisation, even at low temperatures. These results are relevant in the explanation of the origin of detergent-resistant membrane fragments as found, e.g. in caveolae or 'rafts'. PMID- 9733940 TI - GABAa receptor-mediated field potentials are enhanced in area CA1 following prenatal cocaine exposure. AB - Prenatal cocaine exposure results in several documented changes in neurotransmitter receptor number and structure. Increases have been reported for cortical catecholamine and indoleamine receptor number and binding affinity, in the subunit expression of glutamatergic NMDA and AMPA receptors in the striatum, and in GABA immunoreactivity in the anterior cingulate cortex. We sought information on the functional consequences of cocaine-induced alterations in receptor structure/number. Since hippocampal amino acid neurotransmitters are of critical importance and have been shown to be affected by cocaine, we studied field potentials produced by synaptic activation of isolated glutamatergic NMDA and AMPA receptors and GABAa and GABAb responsive receptors in area CA1 of rabbit hippocampal slices. We found the GABAa receptor population produced significantly larger field potentials in cocaine-exposed offspring compared to controls, while other receptors produced responses similar to controls. PMID- 9733941 TI - Genotoxicity in workers exposed to methyl bromide. AB - To address the genotoxicity of in vivo methyl bromide (CAS 74-83-9) exposure in humans, we collected blood and oropharyngeal cells as part of a cross-sectional morbidity study of methyl bromide-exposed fumigation workers and their referents. Micronuclei were measured in lymphocytes and oropharyngeal cells, and hypoxanthine-guanine phosphoribosyl transferase gene (hprt) mutations were measured in lymphocytes. A total of 32 workers and 28 referents provided specimens. Among current non-smokers, mean hprt variant frequencies (Vfs) were found to be elevated among workers compared to referents (geometric mean: workers=4.49x10(-6), referents=2.96x10-(6); two-sided p=0.22); this difference was more pronounced among workers with 4 h or more of recent methyl bromide exposure compared to referents (geometric mean: workers=6.56x10(-6), referents=2.96x10(-6); two-sided p=0.06). Mean oropharyngeal cell micronuclei were higher among workers compared to referents (mean: workers=2.00, referents=1.31; two-sided p=0.08); the results were similar when workers with 4 h or more of recent methyl bromide exposure were compared to referents (mean: workers=2.07, referents=1.31; two-sided p=0.13). No consistent differences between workers and referents were observed for frequencies of kinetochore negative lymphocyte micronuclei, or kinetochore-positive lymphocyte micronuclei. The study was limited by a sample size sufficient only for detecting relatively large differences, absence of a reliable method to measure the intensity of workplace methyl bromide exposures, and relatively infrequent methyl bromide exposure (e.g., the median length of exposure to methyl bromide during the 2 weeks preceding the survey was 4 h). In conclusion, our findings provide some evidence that methyl bromide exposure may be associated with genotoxic effects in lymphocytes and oropharyngeal cells. Further study on the genotoxicity of methyl bromide exposure in humans is warranted. PMID- 9733942 TI - Phosphorylation of the GABAA receptor gamma2L subunit in rat sensory neurons may not be necessary for ethanol sensitivity. AB - The effect of ethanol on the current activated by 2.5 to 40 microM gamma aminobutyric acid (GABA) was studied in freshly isolated rat dorsal root ganglion (DRG) neurons under voltage clamp in the whole-cell and perforated-patch recording configurations. Our results confirmed that GABAA-activated current in these neurons was insensitive to ethanol at concentrations from 2.5 to 100 mM [G. White, D.M. Lovinger, F.F. Weight, Ethanol inhibits NMDA-activated current but does not alter GABA-activated current in an isolated adult mammalian neuron, Brain Res. 507 (1990) 332-336.]. In addition, the ethanol sensitivity of GABA receptors was studied under conditions that promote phosphorylation of the PKC site on the gamma2L subunit. The presence of the gamma2L and other subunit mRNAs was detected by reverse transcription (RT) of total RNA purified from adult DRG followed by polymerase chain reaction (PCR) using subunit specific primer sets. We found that the GABA response remained insensitive to 2.5-100 mM ethanol despite: (i) the extracellular preapplication of 5, 20 or 500 nM phorbol 12 myristate 13-acetate (PMA); (ii) raising free intracellular Ca2+ ([Ca2+]i) from 7 to 100 or 600 nM by altering the intracellular Ca2+/EGTA ratio; (iii) intracellular application of PKC (0.247 U ml-1 ); and (iv) combining the intracellular application of 1 microM okadaic acid and 30 microM peptide 3 with the extracellular application of 20 nM PMA. These results suggest that phosphorylation of the gamma2L subunit is not the only requirement for ethanol sensitivity of GABAA receptors. PMID- 9733943 TI - Cholesterol-dependent generation of a unique amyloid beta-protein from apically missorted amyloid precursor protein in MDCK cells. AB - To investigate the implications of altered sorting of the beta-amyloid precursor protein (betaAPP) in the abnormal generation of amyloid beta-protein (Abeta), we characterized Abeta secreted from Madin-Darby canine kidney (MDCK) cells which had been stably transfected with a cDNA encoding the human beta-amyloid precursor protein (betaAPP695) with a 42 amino acid residue truncation at the carboxyl terminus (DeltaC). In DeltaC MDCK cells, the intracellular sorting of betaAPP is substantially altered to the apical surface. We detected an accumulation of a unique Abeta species in the apical compartment of DeltaC MDCK cell cultures. This unique Abeta was immunoprecipitated with 4G8 (a monoclonal antibody specific for Abeta17-24) and detected as a smear on Western blots, but was not immunoprecipitated with BAN50 (a monoclonal antibody raised against Abeta1-16). Interestingly, however, this Abeta species was readily immunoprecipitated with BAN50 upon treatment with formic acid. Furthermore, incubation of the DeltaC MDCK cells with compactin, an inhibitor of de novo cholesterol synthesis, or with filipin, a cholesterol-binding drug, resulted in marked changes in the characteristics of this Abeta species as follows: first, the Abeta was not observed as a smear on Western blots and second, the Abeta was immunoprecipitated with BAN50. The present results strongly suggest that an Abeta with unique molecular characteristics is generated from the missorted betaAPP in vivo in a cholesterol-dependent manner. PMID- 9733944 TI - The small-eye mutation results in abnormalities in the lateral cortical migratory stream. AB - Mice with homozygous mutations of the Pax-6 gene exhibit a constellation of developmental problems including the absence of eyes and nasal cavities and problems in the movement of neuroblasts out of the germinal epithelium. In this paper, we demonstrate further disturbances in neuronal migration. Normally, cells produced along the lateral ventricles move laterally across the pallium, ultimately coming to reside in the lateral neocortex and primary olfactory cortex. In mutant animals, these cells continue to migrate to the pial surface of the brain. PMID- 9733945 TI - Cholinergic cell expression in the developing rat medial septal nucleus in vitro is differentially controlled by GABAA and GABAB receptors. AB - The early appearance and relative abundance of GABAergic neurons in basal forebrain cholinergic nuclei like the medial septum suggest that the maturation of the later developing cholinergic neurons in these nuclei may be controlled by GABA. To examine this possibility, the effects of both exogenous GABA and specific GABA receptor agonists, as well as that of endogenous GABA on the phenotypic expression and survival of the cholinergic neurons in primary cultures from the fetal rat medial septum, were studied. Treatment of these cultures for six days with GABA significantly decreased the enzymatic activity of choline acetyltransferase (EC 2.3.1.6) (ChAT) in a dose-dependent manner. This response to exogenous GABA was blocked by bicuculline, mimicked by muscimol and slightly potentiated by saclofen. Consistent with this latter observation, the GABAB receptor agonist, baclofen, dose-dependently increased septal ChAT activity. However, while the effect of baclofen on cholinergic expression was lost in the absence of glia, the suppressive effects of GABA or muscimol were more marked. Acetylcholinesterase (EC 3.1.1.7) (AChE) expression in mixed neuronal-glial cultures, was, like ChAT activity, increased or decreased in intensity with the inclusion of baclofen or muscimol, respectively. Although the number of AChE positive neurons in muscimol-treated cultures was significantly lower than that in controls, no changes in neither neuronal nor general cell viability were noted. Finally, as GABAA or GABAB receptor antagonists bicuculline and picrotoxin or saclofen, when applied alone to mixed cultures, increased or decreased ChAT activity, respectively, it appears that endogenous GABA, tonically released in the developing septum, may, via specific receptor types, differentially control the biochemical maturation of the cholinergic neurons. PMID- 9733946 TI - Effects of preweanling chronic naltrindole administration on stress-induced antinociceptive responses in rats. AB - The effect of a daily injection of the delta-selective opioid antagonist naltrindole (1 mg/kg), from birth to postnatal day 19, on the development of stress-induced-antinociception (SIA) and on the antinociceptive response to the mu-selective agonist alfentanil (65 microg/kg) in female rats was investigated. Functional blockade of the delta-receptor during the preweanling period markedly reduced the antinociceptive response to swim-stress in 25-day-old rats, and SIA was only mediated by delta-receptors at this age. In 20-day-old rats and in adults, SIA was predominantly mu-receptor mediated and unaffected by delta receptor blockade. The lack of interference with mu-receptor function was confirmed as alfentanil responses were unaffected by preweanling naltrindole treatment. The data show independence of mu- and delta-receptors in the control of SIA during development and an impairment of delta- but not mu-mediated SIA after chronic delta-antagonist treatment. PMID- 9733947 TI - Comparative mutagenicity of 7H-dibenzo[c,g]carbazole and two derivatives in MutaMouse liver and skin. AB - 7H-Dibenzo[c,g]carbazole (DBC) is an environmental pollutant that produces DNA adducts and tumors in mouse liver and skin following subcutaneous injection and topical application. The two synthetic derivatives 5,9-dimethyl-DBC (DMDBC) and N7-methyl-DBC (NMDBC) induce tissue-specific lesions. DNA adducts and tumors are observed only in liver following exposure to DMDBC and only in skin following exposure to NMDBC. We used the positive selection MutaMouse model to measure the induction of mutations in the two target organs, 28 days after a single subcutaneous injection or topical application of DBC, DMDBC and NMDBC. In liver, DBC and DMDBC induced 30- to 50-fold increases in mutant frequency (MF), while NMDBC had only a weak effect, regardless of the route of administration. After topical application, DBC and NMDBC produced 3.4- to 7.9-fold increases in MF in skin, while DMDBC had a weak effect. After subcutaneous injection, the three compounds had no or weak effect in skin. This study shows gene mutations arise in the respective target organs in which primary DNA damage and tumors are observed. These results illustrate the relevance of the MutaMouse model for testing organ specific mutagens. PMID- 9733948 TI - Differences in delta opioid receptor antinociception, binding, and mRNA levels between BALB/c and CXBK mice. AB - Mu and delta opioid receptors have been demonstrated to mediate supraspinal opioid antinociception. Whereas the recombinant inbred CXBK mouse is notably deficient in mu opioid receptor antinociception, binding density, and mRNA (MOR 1) levels, little is known about delta opioid receptor processes in this strain. The present study thus compared CXBK mice and their BALB/c strain progenitors with respect to delta opioid antinociception, whole-brain receptor binding levels, and mRNA (DOR-1) levels. Following intracerebroventricular injections of the selective delta1 and delta2 opioids DPDPE and [d-Ala2]deltorphin II, respectively, CXBK mice displayed relatively lower antinociception on the tail flick test, resulting in significantly increased ED50 values for both agonists in this strain. Decreased whole-brain specific binding of [3H][d-Ala2]deltorphin II, but not [3H]DPDPE, was also observed in CXBK mice. Solution hybridization with a probe for the DOR-1 revealed increased transcript levels in the caudate-putamen, frontal cortex, and spinal cord of this strain. The present data demonstrate a deficiency in delta1 and delta2 opioid antinociception in CXBK mice concomitant with reductions in whole-brain delta2 receptor binding and regional increases in DOR-1. Whether these observations are causally related remains to be clarified. PMID- 9733949 TI - Non-equivalent cooperation between the two nucleotide-binding folds of P glycoprotein. AB - To identify the roles of the two nucleotide-binding folds (NBFs) in the function of human P-glycoprotein, a multidrug transporter, we mutated the key lysine residues to methionines and the cysteine residues to alanines in the Walker A (WA) motifs (the core consensus sequence) in the NBFs. We examined the effects of these mutations on N-ethylmaleimide (NEM) and ATP binding, as well as on the vanadate-induced nucleotide trapping with 8-azido-[alpha-32P]ATP. Mutation of the WA lysine or NEM binding cysteine in either of the NBFs blocked vanadate-induced nucleotide trapping of P-glycoprotein. These results suggest that if one NBF is non-functional, there is no ATP hydrolysis even if the other functional NBF contains a bound nucleotide, further indicating the strong cooperation between the two NBFs of P-glycoprotein. However, we found that the effect of NEM modification at one NBF on ATP binding at the other NBF was not equivalent, suggesting a non-equivalency of the role of the two NBFs in P-glycoprotein function. PMID- 9733950 TI - Mechanical activity is necessary for the elimination of polyneuronal innervation of developing rat soleus muscles. AB - During early development, rat soleus muscle fibres are innervated by several axons. Neuromuscular activity is involved in the elimination of all but one terminal, but it is not clear whether electrical or mechanical activity is important. Here, we reduced mechanical activity only, by interfering with excitation-contraction coupling. Muscles treated with dantrolene sodium at 9 days produced significantly less force at 13 days of age than normal muscles, and their sensitivity to ACh was greater than that of controls. The elimination of polyneuronal innervation occurs between days 9-12, but in muscles treated with dantrolene, the loss of synapses was slower. Thus, reducing mechanical activity by interfering with excitation-contraction coupling, (a) delays muscle development and (b) reduces the rate of elimination of polyneuronal innervation. PMID- 9733951 TI - Ontogeny of vesicular monoamine transporter mRNAs VMAT1 and VMAT2. I. The developing rat central nervous system. AB - We used in situ hybridization histochemistry to study the expression of the mRNA of the two vesicular monoamine transporters (VMAT1 and VMAT2) during embryonic and postnatal development of the central nervous system (CNS) in the rat. In the adult rat, VMAT2 mRNA is present exclusively in monoaminergic cell groups of the CNS and VMAT1 mRNA was reported to be present in the adrenal medulla and certain intestinal epithelial cells. In contrast to the above, the expression of VMAT1 mRNA has previously never been detected in the central nervous system. This study shows the first evidence that both transporter molecules are expressed in CNS during ontogenesis. We here demonstrate four main expression patterns detected during development: 1. VMAT2 mRNA expression in monoaminergic neurons of the brainstem beginning as early as embryonic day E13. 2. Expression of VMAT2 mRNA in all major sensory relay nuclei of central nervous system. 3. Co-expression of VMAT1 and VMAT2 mRNA in most limbic structures, basal ganglia, as well as in some hypothalamic nuclei. 4. Exclusive expression of VMAT1 mRNA in the neocortical subventricular zone, in the amygdala at early (E15-18) and late (P1-P28) timepoints, the granular cell layer of cerebellum, and in several brainstem motor nuclei. Based on their distribution during development we suggest that monoamines, released in a controlled fashion, might affect wiring of sensory and also motor circuits. VMAT1 mRNA expression may reflect a specific effect of monoamines in glial differentiation and cerebellar granule cell migration and/or differentiation. PMID- 9733952 TI - Functional modifications of alamethicin ion channels by substitution of glutamine 7, glycine 11 and proline 14. AB - Alamethicin is a 20 amino acid, potentially helical peptaibol which forms voltage dependent ion channels in bilayer systems. Two aspects of alamethicin structure have been suggested to be of particular functional significance for stabilization of alamethicin channels. (i) Proline 14 inducing a helix kink is together with glycine at position 11 responsible for an appropriate orientation of the molecules in the conducting associates. (ii) Glutamine 7 lining the channel interior is assumed to stabilize the channel structure by forming inter-helix hydrogen bonds. The functional importance of these residues was probed in macroscopic and single-channel experiments with alamethicin analogs containing polar, side chain bearing residues at position 11 (glutamine, asparagine) or at position 14 (glutamine). In order to investigate the crucial role of glutamine 7 for the stabilization of channel aggregates, this residue was substituted by alanine. The conformation of the lipid bound peptides was determined by circular dichroism spectroscopy. The results show that glutamine 7, glycine 11 and proline 14 are not essential for channel formation but substitution of any residue reduced the number of conductance levels and significantly reduced their lifetimes. Channel stabilization by the introduction of residues with potential hydrogen bonding capacity at positions 11 and 14 was not observed. Differences in the conformation of the lipid bound peptides, their orientation in the bilayer and their affinity for the lipid membrane appear thus to contribute to the modulation of functional properties. PMID- 9733953 TI - Anticonvulsant and glutamate release-inhibiting properties of the highly potent metabotropic glutamate receptor agonist (2S,2'R, 3'R)-2-(2',3' dicarboxycyclopropyl)glycine (DCG-IV). AB - The anticonvulsant effects of intracerebral administration of the highly potent group II metabotropic glutamate receptor agonist, DCG-IV, were tested in fully kindled rats following daily electrical stimulation of the basolateral amygdala. The agonist caused a dose-dependent increase in the generalized seizure threshold (GST) of these seizure susceptible animals within the dose range tested (0. 01 1.0 nmol). The estimated GST100 value (dose causing a 100% increase in GST) for this effect was 0.22 nmol. The anti-seizure activity of DCG-IV was fully inhibited in the presence of the group II metabotropic glutamate receptor antagonist (2S,1'S, 2'S)-2-methyl-2-(carboxycyclopropyl)glycine (MCCG; 40 nmol), while MCCG alone showed no significant inhibitory effect on seizure activity. DCG IV also powerfully inhibited depolarization-induced release of [3H]D-aspartate from rat cerebrocortical synaptosomes, with an IC50 value of 0.39 microM. In this respect, DCG-IV was approximately 70-fold more potent than the clinically effective anticonvulsant drug lamotrigine (IC50=27.7 microM), a proposed neurotransmitter release inhibitor known to inhibit glutamate release, also tested in this assay. These findings demonstrate the high potency of DCG-IV as an anticonvulsant agent and confirm a key role for group II metabotropic glutamate receptors in the control of seizure activity via their modulatory action on neuronal glutamate release. PMID- 9733954 TI - Evaluation of the mutagenicity and antimutagenicity of forty-two 3-substituted flavones in the Ames test. AB - The mutagenic and antimutagenic activities of forty-two synthetic flavones were assessed by the Ames test. The tested flavones included twenty-three 3 nitroflavones, eighteen 3-aminoflavones and the 3-chloroflavone. The mutagenicity was evaluated with Salmonella typhimurium TA100 and YG1042 (an overproducing nitroreductase and O-acetyltransferase TA100 strain) with and without metabolic activation (S9 mix). The antimutagenicity of the non mutagenic derivatives was evaluated against 11 known reference mutagens. A total of 39 synthetic flavones were mutagenic. The mutagenic activities ranged from 0.1 rev/nmole (4'-chloro-6 methoxy-3-nitroflavone) to 6240 rev/nmole (4'-methoxy-3, 3'-diaminoflavone). Two differences were found between the 3-amino and the 3-nitroflavones: (i) the mutagenicity of the 3-aminoflavones required the presence of the metabolic activation; (ii) the 3-amino derivatives were more mutagenic than their 3-nitro counterparts. Increased mutagenicity, as assessed with strain YG1042, was limited to 17/39 derivatives. The mutagenic activity was induced by the presence of the double bond at the 2,3-position for conjugation of the lone-pair electron with the carbonyl group on the 'C' ring. This mutagenicity was modulated by substituents at the 2'-position. Additional mutagenicity was brought by the aminoaromatic and nitroaromatic group reduction by bacterial nitroreductases and by the S9 mix; it was modulated by different substituents on the aromatic rings of the flavones. Three flavones: 3-chloroflavone (1C), 4'-hydroxy-3-nitroflavone (23N) and 2',3-diaminoflavone (2A) showed antimutagenic properties. Compound 1C was efficient against benzo(a)pyrene (BaP), 2-aminofluorene (2AF), 2 aminoanthracene (2AA), 4-nitroquinoline-1-oxide (4NQO) and 1-methyl-3'-nitro-1 nitrosoguanidine (MNNG). Compound 23N inhibited the mutagenicity of BaP and MNNG. The antimutagenic activity of 2A was limited to MNNG. PMID- 9733955 TI - Neuronal supernumerary and dendritic sprouting of the nucleus ambiguus after chronic alteration of peripheral targets in cats. AB - Anatomic changes of neuronal profiles in response to chronic alteration of peripheral targets were investigated in the nucleus ambiguus (NA) of cats. Unilateral vagal-hypoglossal nerve anastomosis was performed by suturing the transected proximal stump of the vagus nerve to the transected distal stump of the hypoglossal nerve. After comparing horseradish peroxidase (HRP)-labeled neurons on the ipsilateral operated side of the NA with the contralateral unoperated NA and the NA following transection and reuniting to the vagus itself, a remarkable ramification and elongation of the dendritic trees was observed in the HRP-positive neurons on the ipsilateral NA. Quantitative analysis of neuronal profiles revealed that the number of the medium and large neurons on the ipsilateral NA was greater than the contralateral NA and the NA following autologous suturing of the vagus. Comparisons of variable dendritic lengths of the medium and large neurons on the ipsilateral NA revealed longer distances and more branches of the tertiary and perisomatic dendrites than those of the contralateral NA and the NA ipsilateral to autologous reunion. Our results suggest that remarkable sprouting and elongation of the dendritic trees as well as cell supernumerary occurred in the dominant NA motoneurons ipsilateral to the nerve anastomosis. In conclusion, there is a trophic influence in the tongue musculature, which was retrogradely transported to the NA neurons via the regenerating axons and caused the morphological changes in the NA in response to the rerouting of efferents from the vagus nerve to the hypoglossal nerve to innervate intimate tongue musculature. PMID- 9733956 TI - Aggregation of dimyristoylphosphatidylglycerol liposomes by human plasma low density lipoprotein. AB - Turbidity (absorbance at 470 nm) measurements revealed human serum low density lipoprotein (LDL) to cause, within a few minutes and at physiological pH and [NaCl], the aggregation of liquid crystalline large unilamellar liposomes (LUVs) of dimyristoylphosphatidylglycerol (DMPG). No evidence for concomitant lipid or aqueous contents mixing was obtained with fluorescent assays for these processes, in keeping with the lack of fusion of LUVs. Involvement of apoB is implicated by the finding that tryptic digestion of LDL abrogates its ability to cause aggregation. Aggregation is not caused by VLDL, HDL2, or HDL3. Interestingly, also oxidised LDL failed to aggregate DMPG vesicles. Aggregation of DMPG LUVs by LDL did depend on the ionic strength of the medium as well as on the phase state of the lipid. More specifically, below the main transition temperature Tm maximal aggregation was seen in the presence of 25-100 mM NaCl, whereas slightly higher (up to 150 mM) [NaCl] were required when T>Tm. Aggregation due to LDL was also observed for dimyristoylphosphatidylserine as well as for dipalmitoylphosphatidylglycerol LUVs, whereas liposomes composed of either unsaturated acidic phospholipids or different phosphatidylcholines were not aggregated. Involvement of electrostatic attraction between the acidic phosphate of DMPG and cationic residues in apoB is suggested by the finding that increasing the content of dimyristoylphosphatidylcholine (DMPC) in DMPG liposomes reduced their aggregation and at XDMPC=0.50 no response was evident. Notably, increasing the mole fraction of 1-palmitoyl-2-oleyl-PG (POPG) in DMPG LUVs progressively reduced their aggregation by LDL and at XPOPG=0.50 there was complete inhibition. The latter effect of POPG is likely to be due to augmented hydration of the unsaturated lipid constituting a barrier for the contact between apoB and the vesicle surface. In keeping with this view, the presence of the strongly hygroscopic polymer, poly(ethylene glycol) at 1% (by weight) enhanced the aggregation and could partly reverse the inhibition by POPG. PMID- 9733957 TI - Treatment with the spin-trap agent alpha-phenyl-N-tert-butyl nitrone does not enhance the survival of embryonic or adult dopamine neurons. AB - Reactive oxygen species are thought to be involved in the death of dopaminergic neurons in Parkinson's disease as well as in transplanted embryonic dopaminergic neurons. The spin-trap agent alpha-phenyl-N-tert-butyl nitrone (PBN) reacts directly with radical species and may thereby prevent them from damaging important cellular molecules such as membrane lipids. We found that PBN does not increase the survival of cultured embryonic dopaminergic neurons subjected to serum deprivation, whereas the antioxidant and lipid peroxidation inhibitor lazaroid U-83836E does. Moreover, PBN does not increase the survival of grafted embryonic dopaminergic neurons or graft efficacy (monitored as changes in drug induced motor asymmetry in hemiparkinsonian rats) when the spin-trap agent is given intraperitoneally to the graft recipient or is added to the solutions used when preparing tissue for transplantation. Another spin-trap agent, alpha-(4 pyridyl-1-oxide)-N-tert-butyl nitrone (POBN) also failed to protect neurons when given to graft recipients in the same experimental paradigm. Finally, we found that adult nigral neurons subjected to a progressive retrograde 6-OHDA lesion are not protected by systemic treatment with PBN. Even though reduction of oxidative stress by overexpression of superoxide dismutase or addition of lazaroids have previously been shown to enhance the survival of cultured and grafted dopaminergic neurons, spin-trap agents PBN and POBN do not provide protection in these experimental paradigms. This may be due to antioxidants and spin-trap agents interfering in different steps of free radical-induced cell damage. PMID- 9733958 TI - Ontogeny of vesicular monoamine transporter mRNAs VMAT1 and VMAT2. II. Expression in neural crest derivatives and their target sites in the rat. AB - We used in situ hybridization histochemistry to study the expression of the two vesicular monoamine transporters (VMAT1 and VMAT2) during embryonic development in the rat. In the adult rat VMAT2 is present exclusively in neuronal tissues and VMAT1 is present in the adrenal medulla and in certain intestinal endocrine cells. We found that both transporter molecules are more widely expressed during development. We demonstrate a complete overlap of the two VMAT mRNAs in the sympathetic nervous system between E13 and E21 days. In addition, VMAT2 (and to some extent VMAT1) mRNA is expressed in ganglionic cells of the parasympathetic nervous system and in cranial ganglia (trigeminal, vestibular and spiral ganglia) between E12 and E21. The sensory neurons of the dorsal root ganglia, which are also neural crest derivatives, express VMAT2 mRNA (E11-E21), exclusively. Both VMAT mRNAs are found in the developing GI system, but in different cells. VMAT1 mRNA was detected in organs of the endocrine system (pituitary gland, adrenal gland, testis, seminal vesicle), some connective tissue cells, and the thymus. We observed expression of both VMAT mRNAs in two separate cell groups in the placenta (E8-E10). Based on their distribution during development we suggest that monoamines, released in a controlled fashion, might affect migration and differentiation of neural crest derivatives. PMID- 9733959 TI - Membrane sterol composition modulates the pore forming activity of syringomycin E in human red blood cells. AB - The effect of lipopeptide antifungal agent, syringomycin E (SRE) on the membrane permeability of human red blood cells (RBCs) was studied. SRE added to RBCs above a concentration of 2x106 molecules/cell (50 microgram/ml RBCs) caused a rapid and concentration dependent lysis of a small subpopulation of RBCs; the extent of this lysis remained unchanged as long as 100 min. During this time period the membranes of the unlysed cells had enhanced permeability for ions which was monitored by direct measurement of 86Rb flux. Both the extent of cell lysis and ion transport rate showed linear relationships with SRE concentration demonstrating a random distribution of SRE molecules in red blood cells. The kinetics of the 86Rb efflux suggested pore formation by syringomycin E. The pores had discrete life times and were eventually inactivated. The pores were also a pathway for efflux of monomeric haemoglobin. Alteration of the membrane sterol composition, i.e. depletion of cholesterol by 50% or partial ergosterol substitution of the cholesterol increased the SRE induced membrane permeability for 86Rb by two orders compared to membranes with unaltered sterol composition. This modification of the sterol composition promotes the pore forming activity of this lipopeptide in the membrane. PMID- 9733960 TI - Neuroanatomical patterns of fos-like immunoreactivity induced by a palatable meal and meal-paired environment in saline- and naltrexone-treated rats. AB - Opioid antagonists block the positive hedonic response to food taste and are potent inhibitors of palatability-driven feeding. However, the specific brain regions within which opioid peptide secretion contributes to the maintenance of palatability-driven feeding have not been clearly established. In the present study, c-Fos immunohistochemistry was used to identify regions rostral to the hindbrain that display cellular activation in response to a palatable meal and the meal-paired environment. Further, it was determined whether any of the cellular responses could be prevented by pretreating animals with naltrexone. Twenty brain regions known to be involved in gustation, appetite and reward functions were examined. Ingestion of the palatable meal (3.0 g of 30% shortening, 20% sucrose and 50% powdered Purina rat chow) increased Fos-like immunoreactivity (FLI) in lateral hypothalamus (LH), ventral tegmentum (VTA) and medial preoptic area (MPOA), and decreased FLI in the habenula (Hab). The meal paired environment increased FLI in the VTA and nucleus accumbens shell (NAC shell). Naltrexone (1.0 mg/kg, i.p.) did not block consumption of the small meal but did prevent all of the distinctive increases in FLI induced by the meal and meal-paired environment. Since naltrexone, alone, increased FLI in VTA, NAC shell, central amygdala (ceA) and laterodorsal bed nucleus of the stria terminalis (BSTLD), the blunting of ingestion reward by naltrexone may result from direct or transsynaptic activating effects on opponent neuronal activity within this highly interconnected set of structures that mediate and modulate reward. PMID- 9733961 TI - Changes of the membrane potential profile induced by verapamil and propranolol. AB - The effects of the organic calcium channel blocker verapamil and the beta receptor blocker propranolol on dipole (phi(d)) and surface (phi(s)) potentials of bilayer lipid membranes were studied. The boundary potentials (phi(b)= phi(d) + phi(s)) of black lipid membranes, monitored by conductance measurements in the presence of nonactin and by capacitive current measurements were compared with phi(s) calculated from the electrophoretic mobility of lipid vesicles. It was shown that the increase of boundary potential, induced by the adsorption of the positively charged propranolol, was caused solely by an increase in surface potential. Although phi(s) also increases due to the adsorption of verapamil, phi(b) diminishes. A sharp decrease of the dipole potential was shown to be responsible for this effect. From Langmuir adsorption isotherm the dissociation constant Kd of verapamil was estimated. The uncharged form of verapamil (Kd=(0.061+/-0.01) mM at pH 10.5) has a tenfold higher affinity to a neutral bilayer membrane than the positively charged form. The alteration of membrane dipole potential due to verapamil adsorption may have important implications for both membrane translocation and partitioning of small or hydrophobic ions and charged groups of membrane proteins. PMID- 9733962 TI - Structural studies of the H+/oligopeptide transport system from rabbit small intestine. AB - A 127-kDa protein was identified as a component of the H+/oligopeptide transport system in brush-border membrane vesicles from rabbit small intestine by photoaffinity labeling with [3H]cephalexin and further photoreactive beta-lactam antibiotics and dipeptides. Reconstitution of stereospecific transport activity revealed the involvement of the 127-kDa protein in H+-dependent transport of oligopeptides and orally active alpha-amino-beta-lactam antibiotics (Kramer et al., Eur. J. Biochem. 204 (1992) 923-930). H+-Dependent transport activity was found in all segments of the small intestine concomitantly with the specific labeling of the 127-kDa protein. By enzymatic deglycosylation, fragments of Mr 116 and 95 kDa were obtained from the 127-kDa protein with endoglucosidase F and N-glycanase, whereas with endoglucosidase H, a fragment of Mr 116 kDa was formed. These findings indicate that the photolabeled 127-kDa protein is a microheterogenous glycoprotein. Surprisingly, it was found that the solubilized and purified 127-kDa protein showed enzymatic sucrase and isomaltase activity. Inhibition of the glucosidase activities with the glucosidase inhibitor HOE 120 influenced neither H+/oligopeptide transport nor photoaffinity labeling of the 127-kDa protein. With polyclonal antibodies raised against the purified 127-kDa protein, a coprecipitation of sucrase activity and the photolabeled 127-kDa beta lactam antibiotic binding protein occurred. Target size analysis revealed a functional molecular mass of 165+/-17 kDa for photoaffinity labeling of the 127 kDa protein, suggesting a homo- or heterodimeric functional structure of the 127 kDa protein in the brush-border membrane. These findings indicate that the H+/oligopeptide binding protein of Mr 127000 is closely associated with the sucrase/isomaltase complex in the enterocyte brush-border membrane. PMID- 9733963 TI - Peripherin-like immunoreactivity in type II spiral ganglion cell body and projections. AB - Peripherin, an intermediate filament protein, is present in neuronal subpopulations of both peripheral and central nervous systems. The distribution of peripherin was studied in the adult rat cochlea using immunohistochemistry on whole mount material, in cryostat sections and sections of plastic embedded tissue. In the spiral ganglion, peripherin labeling was restricted to the perikarya of a subpopulation of neurons and their peripheral and central processes. Peripherin positive neurons had the following features: (i) they have a large eccentric nucleus, they were often found in a cluster of 2 or 3 cells, (ii) they were often located near the intraganglionic spiral bundle fibers, (iii) they represented roughly 8% of the whole ganglion population and (iv) on the average they had smaller perikarya than non-immunoreactive cells. Immunostaining on semithin plastic sections revealed positive reactivity on Type II ganglion cells, while Type I neurons were negative. Double labeling using peripherin and three neurofilament (NF) subunit antibodies confirmed the presence of both markers within the same spiral ganglion cell type. Type II neurons have been previously documented as the only subpopulation of the spiral ganglion that presents a strong positive NF immunoreactivity within their perikarya. In the organ of Corti, peripherin-positive fibers formed bundles that course beneath the outer hair cells and send branches that end as boutons contacting the outer hair cells. All these characteristics suggest that peripherin-positive cells are Type II neurons, and that peripherin constitutes a reliable marker for this spiral ganglion subpopulation, as well as their peripheral and central processes. PMID- 9733964 TI - Different effects of prolonged isocapnic hypoxia on the carotid body and the glomus cells in the wall of the common carotid artery of the chicken. AB - In the chicken, glomus cells are widely distributed not only in the carotid body but also in the wall of the common carotid artery and around each artery arising from the common carotid artery. Effects of chronic isocapnic hypoxia on the chicken carotid body and the glomus cells in and around the arteries were examined by immunohistochemistry and electron microscopy. In chickens exposed to isocapnic hypoxia for 35 days, three- to four-fold increase of the carotid body volume was induced. Immunoreactivity for tyrosine hydroxylase of glomus cells almost completely disappeared. Dense networks of TuJ1-immunoreactive nerve fibers were unchanged, whereas peptidergic nerve fibers, i.e., substance P-, calcitonin gene-related peptide-, vasoactive intestinal peptide-, galanin- and neuropeptide Y-immunoreactive fibers, were decreased in and around the carotid body. At the electron microscopic level, increased secretory activity of the glomus cells was verified. Mature dense-cored vesicles were markedly decreased, although prosecretory granules were numerous around Golgi complexes. Many immature glomus cells filled with rough endoplasmic reticulum and free ribosomes, also appeared in the carotid bodies of hypoxic chickens. In contrast to the carotid body, the glomus cells located in the wall of the common carotid artery revealed no changes after long-term hypoxia. The cells in the hypoxic chickens, as well as normal controls, expressed intense immunoreactivity for neuropeptide Y, serotonin and chromogranin A. Furthermore, a large number of dense-cored vesicles were distributed throughout the cytoplasm. The glomus cells around each artery arising from the common carotid artery were affected by hypoxia, although the degree of their response to hypoxia varied depending on the locations. PMID- 9733965 TI - Endocytosed ricin and asialoorosomucoid follow different intracellular pathways in hepatocytes. AB - Earlier studies have suggested that fluid phase endocytosis in rat hepatocytes takes place via a clathrin-independent mechanism [1,2]. This observation suggests that a relatively large amount of plasma membrane outside coated pits may be involved in hepatic endocytosis. Ricin, which binds to galactose residues on glycoproteins and glycolipids, has, in this report, been used as a general marker for the plasma membrane of hepatocytes. The endocytosis of ricin was compared with that of asialoorosomucoid (AOM) which is taken up exclusively via clathrin coated pits. Hypertonic medium has been shown to inhibit uptake via coated pits more effectively than clathrin-independent uptake [3-5]. It was found, in this study, that the addition of 100 mM sucrose to the incubation medium inhibited the uptake of 125I-tyramine-cellobiose-asialoorosomucoid (125I-TC-AOM) more extensively than that of 125I-tyramine-cellobiose-ricin (125I-TC-ricin), compatible with the notion that the two probes are internalised via different mechanisms. Subcellular fractionation experiments indicated that 125I-TC-ricin entered a denser endocytic organelle than that receiving 125I-TC-AOM. To determine whether the separation of the two probes was due to a different transport kinetics (i.e. that 125I-TC-ricin is transported more rapidly to a later, denser compartment than 125I-TC-AOM) the cells were incubated at 18 degreesC to allow a slower internalisation/transport of the labelled probes. The results obtained showed, again, that the early endosomes containing 125I-TC-ricin were significantly denser than those containing 125I-TC-AOM. We also employed the horseradish peroxidase (HRP)-diaminobenzidine (DAB) density shift technique of Courtoy et al. [6] to determine whether 125I-TC-ricin and 125I-TC-AOM were in separate endosomes early after their uptake. The results showed that early endosomes containing 125I-TC-AOM were density shifted whereas those containing 125I-TC-ricin were unaffected by the density shift procedure. The use of probes labelled with 125I-TC allowed us to identify compartments involved in the degradation of 125I-TC-AOM and 125I-TC-ricin, by measuring acid soluble radioactivities in the gradient fractions. It was found that 125I-TC-ricin was degraded mainly in endosomes, whereas 125I-TC-AOM, as expected, was degraded mainly in lysosomes. PMID- 9733966 TI - Restriction of environmental space attenuates locomotor activity and hippocampal acetylcholine release in male rats. AB - We examined the effects of the restriction of environmental space on hippocampal acetylcholine release and spontaneous locomotor activity. Four days after the housing in a large or small cage, sampling for microdialysis study was begun. The locomotor activity counts exhibited significant daily changes in all rats in either the large or small cage. But, the mean locomotor activity counts in rats in the small cage was significantly less than that in the large cage. In contrast, the amount of acetylcholine collected per 20-min sample exhibited significant diurnal changes in all six rats in the large cage and in 5 of 6 rats in the small cage. The mean acetylcholine release in the rat in the small cage was significantly lower than that in the rat in the large cage during the dark phase, but not during the light phase. In addition, during the dark phase, hippocampal acetylcholine release was closely associated with spontaneous activity in all six rats in the large cage but not in 3 of 6 rats in the small cage. The present study suggests that the restriction of environmental space somehow interfere with the spontaneous locomotor activity and hippocampal acetylcholine release during the dark phase. PMID- 9733967 TI - The phase behavior of aqueous dispersions of unsaturated mixtures of diacylglycerols and phospholipids. AB - The phase behavior of mixtures of 1-palmitoyl-2-oleoyl-sn-glycerol (1,2-POG) with 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and 1-palmitoyl-2-oleoyl sn-glycero-3-phosphoserine (POPS) was studied by using DSC, small-angle X-ray diffraction and 31P-NMR. The results have been used to construct phase diagrams for both type of mixtures, in the 0-45 degreesC range. It is concluded that 1, 2 POG form complexes in the gel phases with both POPC and POPS. In the case of POPC, two complexes are postulated, the first one at a 1, 2-POG/POPC molar ratio of 40:60, and the second one at 70:30, defining three different regions in the phase diagram. Two eutectic points are proposed to occur: one at a very low 1,2 POG concentration and the other at a 1,2-POG concentration slightly lower than 70%. In the case of the 1,2-POG/POPS mixtures, the pattern was similar, but the first complex was seen to happen at a higher concentration, about 50 mol% of 1,2 POG, whereas the second was found at 80 mol% of 1,2-POG. This indicated a bigger presence of 1,2-POG in the complexes with POPS than with POPC. In the first region of the phase diagram, i.e. at concentrations of 1,2-POG lower than that required for the formation of the first complex, and at temperatures above the phase transition, lamellar phases were seen in all the cases. In region 2 of the phase diagram, i.e. at concentrations where the first and the second complexes coexist, a mixture of lamellar and non-lamellar phases was observed. Finally, at high concentrations of 1,2-POG, non-lamellar phases were detected as predominant, these phases being of an isotropic nature, according to 31P-NMR. An important conclusion of this study is that, using unsaturated lipids, similar to those found in biological membranes, it has been shown that diacylglycerols are found separated in domains, and that this process starts at very low concentrations of diacylglycerols. The formation of separated domains enriched in diacylglycerol is biologically relevant as it will allow them to have important effects on the membrane structure besides the fact that their concentration in the biomembrane is relatively low. PMID- 9733968 TI - Is there a geniculohypothalamic tract in primates? A comparative immunohistochemical study in the circadian system of strepsirhine and haplorhine species. AB - In rodents, the circadian rhythm generated by the hypothalamic suprachiasmatic nucleus (SCN) is modulated by two types of phenomena: photic phase-shifts, mediated by the retinohypothalamic pathway and non-photic phase-shifts mediated by the projection of the intergeniculate leaflet (IGL) to the SCN which contains the neuropeptide Y (NPY). In primates, the retinohypothalamic pathway has been well-demonstrated but very little is known about the geniculohypothalamic tract. This prompted us to study NPY immunoreactivity in both the SCN and the IGL in species representative of the three main primate lineages: prosimians (Microcebus), New World monkeys (Callithrix) and Old World monkeys (Macacca). In species studied, we found a region in the pregeniculate nucleus containing both NPY immunopositive cells and substance P immunopositive fibres that we identified as the IGL. During evolution, this structure has moved from a ventral to a dorsomedial position relative to the adjacent dorsal lateral geniculate nucleus. By contrast, NPY-IP fibres in the SCN are dense in prosimians, but are sparse or absent in other primate species. We suggest that either the geniculohypothalamic projection is absent in higher primates as is the case in humans, or is absent in diurnal mammals, or contains a different peptide, or that NPY immunoreactivity varies according to other parameters. PMID- 9733969 TI - Effect of amphotericin B on dipalmitoylphosphatidylcholine membranes: calorimetry, ultrasound absorption and monolayer technique studies. AB - Amphotericin B (AmB) is a popular drug frequently applied in the treatment of mycosis. Differential scanning calorimetry (DSC), ultrasound absorption and monomolecular layer technique were applied to study the effect of AmB on organisation of dipalmitoylphosphatidylcholine (DPPC) membranes. DSC-determined enthalpy of the main phase transition of DPPC liposomes was found to be a sensitive parameter to monitor AmB-DPPC interaction. The enthalpy of the phase transition decreases with the increase in molar fraction of AmB incorporated to membranes. The exceptionally sharp decrease in the enthalpy of the transition was observed in the membranes containing 5-7 mol% AmB. Ultrasound absorption monitored main phase transition of DPPC is very broad under the presence of 5 mol% AmB showing destabilisation and disorganisation of a membrane structure. These findings are discussed in comparison to monomolecular layer study of two component DPPC-AmB system. Analysis of the surface pressure-molecular area isotherms of compressing DPPC-AmB films at the air-water interface shows pronounced increase in mean molecular area at AmB concentrations corresponding to those found to destabilise DPPC membranes of liposomes. Disorganisation of lipid bilayers due to the presence of AmB in concentrations below 10 mol% with respect to lipid is discussed in terms of toxicity and side effects of this drug. PMID- 9733970 TI - Excitatory effects of muscarine on septohippocampal neurons: involvement of M3 receptors. AB - Cholinergic mechanisms in the septohippocampal pathway contribute to several cognitive functions and impaired cholinergic transmission in this pathway may be related to the memory loss and dementia that accompanies normal aging and Alzheimer's disease and behavioral studies suggest that muscarinic mechanisms in the medial septum/diagonal band of Broca (MSDB) may contribute to these functions. The goal of the present study was to begin a characterization of the physiological and pharmacological effects of muscarine on antidromically identified septohippocampal neurons (SHNs). Muscarinic agonists produced a concentration-dependent excitation in >90% of SHNs tested using extracellular recordings in an in vitro rat brain slice preparation. The SHNs excited by muscarine had a broad range of conduction velocities (0.2 to 3.7 m/s; mean: 1.6+/ 0.06 m/s; n=110), suggesting involvement of neurons with both slow (possibly cholinergic) and fast (possibly GABAergic) conducting fibers. The muscarine induced excitations in SHNs were found not to be mediated via M1, M2 or M4 receptors, as they were not blocked by the M1-selective antagonists, pirenzepine or telenzepine or by the M2/M4-selective antagonist, methoctramine. In contrast, the M3-selective antagonist, 4-DAMP-mustard, blocked muscarinic excitations in a majority of SHNs, indicating the presence of M3 as well as non-M3-type responses. McN-A-343, an M1 and M5-selective agonist, excited 33% of neurons tested, confirming involvement of non-M3 receptors (possibly M5) and M3 receptors. Since the cholinergic and GABAergic MSDB neurons together innervate almost every type of hippocampal neuron, the effects of muscarine on SHNs would also have a profound effect on hippocampal circuitry. PMID- 9733971 TI - Steady-state binding of [3H]ATP to rat liver plasma membranes and competition by various purinergic agonists and antagonists. AB - Steady-state analysis of nucleotide-binding sites on rat liver plasma membranes was carried out using 3H-labelled ATP as radioligand under complete inhibition of ecto-ATPase activity by excess EDTA. Binding of [3H]ATP to the membranes is saturable, reversible and apparently involves one population of specific binding sites with Kd of about 90 nM and binding capacity (Bmax) of 15 pmol/mg protein. A broad spectrum of purinergic agonists and antagonists was examined as potential inhibitors of the measured binding. The displacement studies showed the following rank order of inhibitory potency for [3H]ATP-binding sites (pIC50 values in parentheses): ATPgammaS (7.49)>2-MeSATP (7.18)>ATP (6.91)>ADPbetaS (6.64)>/=ADP (6.56)>>RB2 (6.14)>>suramin (5.40)>>Ap4A (4. 57)>alpha,beta-MeATP (4.19)>/=beta,gamma-MeATP (3.97). AMP, adenosine, Ap5A, PPADS, beta glycerophosphate as well as non-adenine nucleoside triphosphates GTP, UTP and CTP did not exert any effect on the measured binding at concentration ranges of 10-6 10-4 M. In order to ascertain whether ATP and its analogues are capable of interacting with the same binding domain, 2-MeSATP and ADP were treated as alternative ligands that could compete with unlabelled ATP for its binding sites. A 2-fold increase of Kd value for ATP-receptor interaction was observed in the presence of 2-MeSATP (60 nM) or ADP (250 nM) without any modulation of Bmax value, confirming that inhibitory effects of these compounds are competitive in nature. These studies demonstrate that ATP and its analogues are able to interact with a single binding domain on liver plasma membranes, which may be identified as ligand-binding component of P2 purinoceptors of the P2Y1 subtype. PMID- 9733972 TI - Striking changes in anxiety in Huntington's disease transgenic mice. AB - Huntington's disease transgenic mice were tested in the elevated plus-maze test of anxiety at 6, 8, 10 and 12 weeks of age. At all ages, they showed significant and striking increases in the percentages of open arm entries and time spent on the open arms, compared with their normal littermates, indicating reduced anxiety. These increases were not secondary to a non-specific stimulant effect, since the transgenic mice made fewer closed arm entries, significantly so from 10 weeks of age. The mice were also tested in the holeboard, which provides measures of locomotor activity and directed exploration. From 8 weeks of age, the Huntington's mice were significantly less active than their normal littermates and made fewer exploratory head-dips. The increased open arm activity in the elevated plus-maze cannot therefore be secondary to increased exploration in the transgenic mice. In order to determine whether the reduced anxiety was due to differences in benzodiazepine receptor function, the mice were challenged with the benzodiazepine receptor antagonist, flumazenil. The results indicated that some of the reduced anxiety could be attributed to the presence of an endogenous anxiolytic ligand. PMID- 9733973 TI - Isolation and sequence of omcA, a gene encoding a decaheme outer membrane cytochrome c of Shewanella putrefaciens MR-1, and detection of omcA homologs in other strains of S. putrefaciens. AB - The sequence of the omcA gene, which encodes a decaheme cytochrome c that is localized to the outer membrane (OM) of Shewanella putrefaciens MR-1, was determined. The 2202 bp nucleotide sequence of omcA encodes for 734 amino acids with a predicted molecular protein mass of 78.6 kDa. Comparison with the amino terminal sequence of the mature protein suggests the presence of a hydrophobic leader sequence which is cleaved during translocation of the protein to the OM. This leader sequence has a lipoprotein consensus sequence for signal peptidase II at the cleavage site. The predicted mature protein is comprised of 708 amino acids with a predicted molecular mass of 75.8 kDa, but the addition of ten covalently attached heme c groups and covalent lipid modification to the amino terminal cysteine increases the predicted mass to 82.7 kDa. This is consistent with its apparent mass of 83 kDa in SDS-PAGE gels. The predicted amino acid sequence for the OmcA protein shows no significant homology to known proteins. A RNA of approx. 2300 bases that hybridizes to the omcA gene was detected in anaerobically grown MR-1 cells. The size of this transcript is similar to the coding region of the omcA gene, suggesting that it is not part of a multicistronic operon. Similar to MR-1, four other strains of S. putrefaciens were all found to localize a majority of their membrane-bound cytochromes to the OM when grown under anaerobic conditions, and all contained an OM cytochrome of similar size to OmcA. In two of these strains, MR-4 and MR-8, a homolog of omcA was identified by RT-PCR and Southern blotting using primers and probes specific for omcA of MR-1. Western blot analysis using a polyclonal antibody to OmcA was similarly positive in strains MR-4 and MR-8. Partial nucleotide sequence analysis of these homologs demonstrated 74-77% predicted amino acid homology with OmcA of MR-1. In contrast, strains MR-30 and MR-42 tested negative for omcA homologs by Southern and Northern blots, RT-PCR, and Western blots. PMID- 9733974 TI - The protein state of matter. AB - Three ways are generally used to visualize proteins: (1) a static model in which the atomic positions are defined, (2) a dynamic model taking into account fluctuations, and (3) a reactive model that reflects the internal and external electric fields of the molecule. The properties of chromophoric prosthetic groups can be probed by optical spectroscopy, and when high resolution techniques are used, the results reveal information about the local electric fields in proteins, as influenced and determined by atomic positions and dynamics. PMID- 9733975 TI - Serotonin transporters are located on the axons beyond the synaptic junctions: anatomical and functional evidence. AB - The serotonin (5-HT) transporter (5-HTT) is known to play a role in depression and many 5-HT related diseases, and is the target site for drugs of abuse, such as cocaine, MDMA, and methamphetamine. The major role of the 5-HTT has long been considered to be to inactivate serotonin transmission through the elimination of serotonin at release sites. However, immunocytochemistry using an antibody against the N-terminal of the 5-HTT at the light microscopic (LM) level indicates that the 5-HTT is associated not only with 5-HT varicosities but also with axons. Electron microscopy (EM) reveals that the majority of the 5-HTTs exist on the axolemma outside the synaptic junctions. In studying whether axonal 5-HTTs are involved in the uptake of 5-HT, we found with autoradiography that [3H]citalopram bound to all major 5-HT fibers, not only in the terminal regions, but also in 5 HT axonal bundles such as the cingulum bundle and medial forebrain bundle. Furthermore, voltammetry recordings indicated that serotonin axonal bundles were actively engaged in high affinity serotonin uptake. The evidence indicates that 5 HTTs on 5-HT axons away from the synapse are likely to be functional in a manner similar to the terminal 5-HTT for serotonin uptake. It also suggests that the role of the 5-HTT may not only be for the termination of synaptic transmission, but also for the regulation of 5-HT through extrasynaptic (volume) transmission. Our findings may also impact the understanding of the sites of action of selective serotonin reuptake inhibitors and drug entry into serotonin neurons via the numerous axonal sites. PMID- 9733976 TI - Peptide aminonitrogen transport by the lactating rat mammary gland. AB - Recent studies have shown that the lactating mammary gland is able to utilize plasma-derived dipeptides for milk protein synthesis. However, it was not clear whether the peptides were hydrolysed followed by uptake of the constituent amino acids or were taken up intact. In view of this, we have designed experiments to investigate (a) whether the lactating rat mammary gland is capable of transporting hydrolysis-resistant dipeptides and (b) whether or not mammary cells are able to hydrolyse peptides, including glutathione, extracellularly. The uptake of the hydrolysis-resistant dipeptides D-[3H]Phe-L-Gln and D-[3H]Phe-L-Glu by the perfused rat mammary gland was low. Concomitant addition of L-Leu-L-Ala (50 mM) had no effect on the clearance of either labelled dipeptide suggesting that the small, albeit significant, uptake of the dipeptides is not via a high affinity peptide transporter (PepT1/PepT2). All anionic dipeptides tested (L-Glu L-Ala, L-Asp-L-Ala, L-Ala-L-Asp, L-Asp-Gly, Gly-L-Asp and Gly-L-Glu) with the exception of D-Phe-L-Glu were able to trans-accelerate the efflux of labelled D aspartate from preloaded rat mammary tissue (explants and perfused mammary gland). It appears that these peptides were being hydrolysed extracellularly followed by the uptake of free anionic amino acids via the mammary tissue high affinity, Na+-dependent anionic amino acid carrier operating in the exchange mode. Glutathione was able to trans-accelerate D-aspartate efflux from lactating rat mammary tissue in a fashion which was sensitive to the peptidase inhibitor acivicin. This suggests that gamma-glutamyltranspeptidase hydrolyses glutathione to produce L-glutamate which is subsequently transported via the high-affinity anionic amino acid carrier. Hydrolysis of peptides followed by uptake of the constituent amino acids may provide an important source of amino acids for milk protein synthesis. PMID- 9733977 TI - Do all of human midbrain tyrosine hydroxylase neurons synthesize dopamine? AB - We examined whether all of human midbrain tyrosine hydroxylase (TH) neurons substantially synthesize dopamine (DA) using dual labeling immunohistochemical technique of TH and aromatic L-amino acid decarboxylase (AADC). In the substantia nigra, besides many neurons doubly stained for TH and AADC, neurons stained only for TH and only for AADC (D-neurons [C.B. Jaeger, D.A. Ruggiero, V.R. Albert, T.H. Joh, D.J. Reis, Immunocytochemical localization of aromatic l-amino acid decarboxylase, in: A. Bjorklund, T. Hokfelt (Eds.), Handbook of Chemical Neuroanatomy, Classical Transmitters in the CNS, Vol. 2, Part 1, Elsevier, Amsterdam, 1984, pp. 387-408.]) were identified. In the ventral tegmental area, dually labeled neurons and TH-only-positive neurons were found. It is indicated that the number of midbrain TH neurons does not reflect the exact number of DA neurons. PMID- 9733978 TI - Proteins in electric fields and pressure fields: basic aspects. AB - This paper emphasizes the basic aspects of the interactions of chromoproteins at low temperatures with external pressure fields and electric fields. We discuss how the respective spectral properties can be modified and what we can learn from the spectral changes about the thermodynamic, electrostatic, functional and structural properties of proteins. A few examples are discussed in more detail. PMID- 9733979 TI - A novel MPTP primate model of Parkinson's disease: neurochemical and clinical changes. AB - Positron emission tomography (PET) and the dopamine (DA) metabolism tracer, [18F]6-fluoro-L-m-tyrosine (FMT) were used to evaluate the relationship between DA metabolism and the clinical stage of parkinsonism monkeys following either unilateral ICA MPTP infusion or unilateral ICA MPTP infusion and subsequent varying sequential systemic doses of MPTP. Clinical stage corresponded to PET measures of striatal DA metabolism, showing the usefulness of the overlesioned hemiparkinsonian monkey as a stable model of various stages of Parkinson's disease (PD). PMID- 9733980 TI - Molecular and topological characterization of the rat parotid Na+-K+-2Cl- cotransporter1. AB - Na+-K+-2Cl- cotransporters play a central role in driving salt and water movements across secretory and absorptive epithelia. We report the cloning of the rat parotid secretory Na+-K+-2Cl- cotransporter, rtNKCC1. The predicted amino acid sequence of this protein is highly homologous to a previously cloned NKCC1 from the shark rectal gland and to mammalian NKCC1s cloned from several cultured cell lines, confirming the presence of the NKCC1 isoform in a naturally occurring mammalian secretory epithelium. In contrast to previously published NKCC1 clones, our sequence also includes an apparently complete 2680 bp 3'-UTR. Hydropathy analyses of rtNKCC1 predicts that this protein consists of large hydrophilic N and C termini (approx. 30 kDa and 50 kDa, respectively) flanking a central hydrophobic transmembrane region consisting of ten to 12 membrane spanning domains. In addition, we report the results of confocal immunofluorescent microscopic studies using rat parotid acini and antibodies directed against specific regions of the predicted N- and C-terminal portions of rtNKCC1. These studies demonstrate that the epitopes recognized by these antibodies are exposed in permeabilized but not in unpermeabilized cells, indicating that the predicted N and C termini of rtNKCC1 are intracellular. PMID- 9733981 TI - Localization of odor-induced neuronal activity in the antennal lobes of the blowfly Calliphora vicina: a [3H] 2-deoxyglucose labeling study. AB - The distribution of odor-evoked neuronal activity in the antennal lobes of the blowfly Calliphora vicina has been studied by means of [3H] 2-deoxyglucose (2-DG) uptake. Stimulation with natural attractants like meat or cheese induced uptake of 2-DG in a large number of glomeruli, stimulation with the single odorant butyric acid only in a small number of glomeruli. In control brains, the majority of glomeruli showed low 2-DG uptake. The distribution of labeled glomeruli was odor-specific and consistent in different specimen. The labeling patterns evoked by different odors overlapped partially but were clearly distinct. The highest uptake within the glomeruli was found in the axons of antennal receptor neurons. The present data provide evidence that different odors are represented as defined spatial patterns of activity across the antennal lobe glomeruli. The overlapping patterns suggest that certain glomeruli participate in the nervous processing of different odors. PMID- 9733983 TI - The role of inorganic phosphate in regulating the kinetics of inositol 1,4,5 trisphosphate-induced Ca2+ release: a putative role for endoplasmic reticulum phosphate transporters. AB - The effects of phosphate and acylphosphonate phosphate transporter inhibitors were investigated on inositol 1,4,5-trisphosphate (InsP3)-induced Ca2+ release from cerebellar microsomes. Although neither changing the phosphate concentration nor adding phosphate transporter inhibitors affected the percentage (extent) of InsP3-induced Ca2+ release, they did, however, affect the transient kinetics of this process. InsP3-induced Ca2+ release is biphasic in nature, arising from two populations of InsP3-sensitive Ca2+ stores which either release Ca2+ in a fast or slow fashion. Altering phosphate concentration or adding phosphate transporter inhibitors appeared to affect only the fast phase component. We therefore suggest that these observations could be explained by the possibility that phosphate transporters only reside in the fast releasing InsP3-sensitive Ca2+ stores. PMID- 9733982 TI - Neuronal and volume loss in CA1 of the hippocampal formation uniquely predicts duration and severity of Alzheimer disease. AB - In a series of multiple regression models predicting either duration or severity of Alzheimer disease (AD) patients, significant linear correlations were found consistently for the volume of CA1, the subiculum, and the entorhinal cortex. Similarly, the total number of neurons in CA1, CA4, and the subiculum was correlated significantly with both the duration and the severity of AD. A hierarchical multiple regression model was used to examine whether any of these intercorrelated measures had any unique relationship to disease duration or severity. The results showed that only CA1 demonstrated a unique contribution to the explained variance in predicting duration or severity of AD for volume and for neuronal numbers. These results indicate that in the hippocampal formation, volume and neuronal numbers of CA1 appear to show a unique relationship with clinical measures of AD. PMID- 9733984 TI - Proton-coupled oligopeptide transport by rat renal cortical brush border membrane vesicles: a functional analysis using ACE inhibitors to determine the isoform of the transporter. AB - We demonstrate that the angiotensin-converting enzyme inhibitors enalapril and captopril inhibit the transport of D-Phe-L-Gln into PepT1-expressing Xenopus oocytes and into rat renal cortical brush border membrane vesicles (BBMV). The kinetics of inhibition are competitive. Enalapril and captopril are not substrates for PepT2 (Boll et al., Proc. Natl. Acad. Sci. 93 (1996) 284-289). Therefore we conclude that in rat renal cortical BBMV this neutral dipeptide is transported via PepT1. PMID- 9733985 TI - A unifying Tm diagram for phosphatidylethanolamines with sn-1 C20 saturated and sn-2 C18 unsaturated acyl chains. AB - We have determined calorimetrically the phase transition temperature (Tm) values of five sn-1 saturated/sn-2 unsaturated phosphatidylethanolamines (PE) in which the sn-1 acyl chain has 20 carbons and the sn-2 acyl chain has 18 carbons with different number and position of the cis double bond. When these Tm values are combined with the five published Tm values of related unsaturated PE, a unifying Tm diagram is generated for the first time. Moreover, as the molecular mechanics simulated structures of these lipids are taken into consideration, this unifying Tm diagram provides insight into how variations in the number and position of the cis double bond in the lipid's sn-2 acyl chain can influence the phase transition behavior of the lipid bilayer. PMID- 9733987 TI - Proteins in electric fields and pressure fields: experimental results. AB - Experimental results obtained by Stark effect and pressure tuning optical spectroscopy are discussed with the emphasis on studies aimed at unraveling the coupling of prosthetic groups to proteins. A comparative, detailed analysis is given concerning the coupling of the heme group to the apoprotein in various heme proteins based on spectral hole burning data. Electrochromism and electric dichroism experiments related to the coupling problem are also discussed in the context of other protein systems. PMID- 9733986 TI - Light-harvesting complex II in monocomponent and mixed lipid-protein monolayers. AB - Monomolecular layers at the air-water interface were formed directly with isolated largest light-harvesting pigment-protein complex of Photosystem II (LHC II) or out of egg yolk lecithin (EYL) liposomes containing incorporated LHC II. Pure protein monolayers showed a mean area of 1400 A2 per molecule at the air water interface. Monolayers were deposited onto glass slides by means of Langmuir Blodgett (LB) technique. Chlorophyll fluorescence of LHC II-LB and EYL-LHC II-LB films proved energetic coupling of chlorophyll a and b, thus indicating native conformation of LHC II within the monolayers. Scanning force microscopy (SFM) revealed ring-like structures formed in monocomponent protein layers as well as in mixed protein-lipid films. These results suggest that a structural arrangement of LHC II is favoured in a lipid environment but that the protein has itself a strong tendency for structural complex rearrangement in our system. PMID- 9733988 TI - Interfacial indazolization: novel chemical evidence for remarkably high exo surface pH of cationic liposomes used in gene transfection. AB - Cationic liposomes are used as the carriers of polyanionic genes for combating against hereditary diseases in gene therapy. Studies directed to careful biophysical characterizations of the cationic liposomes commonly used in gene delivery have just begun. Herein, we report on a novel liposomal exo-surface bound indazolization reaction of an amphiphilic arenediazonium salt as evidence for the existence of remarkably alkaline exo-surface of cationic liposomes commonly used in gene transfection. Our results demonstrate that formation of 5 hexadecyl-7-methylindazole in thermal indazolization of 2,6-dimethyl-4 hexadecylbenzenediazonium tetrafluoroborate bound to liposome surface is a strong indication for the existence of significantly high exo-surface pH for cationic liposomes commonly used in gene delivery. The present method can be used in determining the relative exo-surface basicities of various cationic liposomes used in gene transfection and subsequently to find any possible correlation between the transfection efficiencies of these liposomes and their exo-surface basicities. PMID- 9733989 TI - Intrinsic protein electric fields: basic non-covalent interactions and relationship to protein-induced Stark effects. AB - Knowledge of the interactions involving charged, polar and polarizable groups in proteins is fundamental, not only because they are important determinants for gaining insight into biophysical molecular recognition and assembly processes, but also for understanding how the matrix of a protein can be viewed as an electric field capable of inducing Stark perturbations on the spectral properties of biological optical centers. This review describes the essential features of noncovalent interactions in protein systems and discusses the concept of the dielectric constant of a protein in the context of different microscopic and macroscopic modeling approaches. It also provides an account of a specific type of high resolution vibrational and optical Stark spectroscopy attempting to correlate the observed spectral properties of biological optical centers to the intrinsic protein fields induced by the matrix in which they reside. PMID- 9733990 TI - High-level expression of Na+/D-glucose cotransporter (SGLT1) in a stably transfected Chinese hamster ovary cell line. AB - The coding region of the high affinity Na+/d-glucose cotransporter (SGLT1) was inserted into the eukaryotic expression vector GFP-N1 under the control of a CMV promoter. The plasmid was then stably transfected into a Chinese hamster ovary cell line (CHO). Transcription and synthesis of SGLT1 were proved by Northern and Western blot analyses. Transport activities of the transfected cells (G6D3) were examined by measuring the sodium-dependent uptake of alpha-methyl[14C]d-glucoside (AMG). Kinetic analysis revealed a Vmax of 10.3 nmol/min/mg (total cell protein) and a Km of 0.26+/-0.09 mM, respectively. The concentration of phlorizin required to inhibit AMG uptake by 50% in the presence of 0.1 mM AMG was 2.35+/-1.84 microM. Electrophysiological studies showed that AMG induces a significant depolarization of membrane voltage in stably transfected CHO cells, suggesting an electrogenic Na-AMG symport. Immunoprecipitation with an antipeptide antibody yielded a nearly homogeneous polypeptide with a molecular mass of about 72 kDa. The amount of SGLT1 present in the CHO cell plasma membranes represents at least 1% of membrane protein, which is about 30-100 times higher than in natural sources, such as renal brush border membranes. In conclusion, the stably transfected G6D3 cells with a markedly high SGLT1 expression can serve as a promising model for studying cellular events related to Na+/d-glucose cotransport and for analyzing the structure and function of the cotransporter itself. PMID- 9733991 TI - Cloning of a cDNA coding for an amino acid carrier from Ricinus communis (RcAAP1) by functional complementation in yeast: kinetic analysis, inhibitor sensitivity and substrate specificity. AB - A cDNA for the amino acid permease gene RcAAP1 has been isolated from Ricinus communis by yeast complementation and subjected to a detailed kinetic analysis. RcAAP1 cDNA is 1.5 kb with an open reading frame that codes for a protein with 486 amino acids and a calculated molecular mass of 53.1 kDa. RcAAP1-mediated histidine uptake was pH dependent with highest transport rates at acidic pH; it was sensitive to protonophores and uncouplers and the Km for histidine uptake was 96 microM. The substrate specificity was investigated by measuring the levels of inhibition of histidine uptake by a range of amino acids. The basic amino acids (histidine, lysine and arginine) showed strongest inhibition of uptake whereas acidic amino acids competed less effectively. Alanine was the most efficient competitor of the neutral amino acids. Glutamine, serine, asparagine, methionine and cysteine showed moderate inhibition whereas threonine, isoleucine, leucine, phenylalanine, tyrosine and tryptophan showed only low levels of inhibition. Glycine, proline and citrulline caused slight stimulation. More detailed competition kinetics indicated that both lysine and arginine showed simple competitive inhibition of histidine uptake. When direct uptake measurements were carried out, both lysine and arginine were found to be effective substrates for RcAAP1. PMID- 9733992 TI - Fluorescence line narrowing applied to the study of proteins. AB - Fluorescence line narrowing is a high resolution spectroscopic technique that uses low temperature and laser excitation to optically select specific subpopulations from the inhomogeneously broadened absorption band of the sample. When applied to the study of fluorescent groups in proteins one can obtain vibronically resolved spectra, which can be analyzed to give information on spectral line shapes, vibrational energies of both the ground and excited state molecule, and the inhomogeneous distribution function of the electronic transitions. These parameters reveal information about the chromophoric prosthetic group and the protein matrix and are functions of geometric strains and local electric fields imposed by the protein. Examples of the use of fluorescence line narrowing are discussed in investigations of heme proteins, photosynthetic systems and tryptophan-containing proteins. PMID- 9733993 TI - Contribution of trifluoperazine/lipid ratio and drug ionization to hemolysis. AB - The interaction of the antipsychotic drug trifluoperazine (TFP) with membranes was investigated in terms of lipid phase perturbation. TFP partition coefficients (P) were measured by phase separation between octanol/water and model membranes/water. The profile of P values at pH 7.4 was: microsomes (7172+/ 1229)>liposomes (1916+/-341)>erythrocyte ghosts (1380+/-429)>octanol (452+/-55). Hemolytic experiments showed a biphasic, protective (at lower concentrations) and hemolytic effect above the CMC (42 microM at pH 7.4) of the phenothiazine. By applying classical treatments for surface active compounds to the hemolytic curves, we could calculate P values in whole erythrocyte cells. The preferential binding of uncharged to charged TFP in the membrane was discussed, since it results in a ionization constant (pKapp) different from that observed in the aqueous phase (pK). The TFP ionization constant was decreased from 8.1 (in water) to 7.62 in the presence of membranes and almost the same ratio of charged/uncharged TFP species is present at physiologic pH. Taking into account the DeltapK, we calculated the average TFP partition coefficient between egg phosphatidylcholine liposomes and water, at pH 7.4 (Paverage=1432), which was well correlated with the measured one (Plip=1916). Paverage is highly influenced by the uncharged TFP species and the real base/acid ratio under physiologic conditions was discussed in terms of its possible role in the biological activity of TFP. PMID- 9733994 TI - Modulation of rat incisor odontoblast plasma membrane-associated Ca2+ with nifedipine. AB - In addition to the Ca2+ portion freely dissociated in the cytosol, another Ca2+ pool is associated with plasma membranes and intracellular organelle membranes. This Ca2+ portion is of importance for regulation of, among other things, the cell cycle, actin-mediated processes, and cell morphology. In the literature, dihydropyridines have been reported to influence this membrane-associated pool of Ca2+ under certain conditions. The aim of this investigation was to study possible modulations of plasma membrane-associated Ca2+ upon treatment with nifedipine in vitro in a Ca2+-transporting cell, the dentin-forming odontoblast. The membrane-associated portion of Ca2+ in dissected dentinogenically active rat incisor odontoblasts was monitored by fluorescence spectrophotometry using chlortetracycline as a probe. In addition, images of chlortetracycline-Ca2+ binding were obtained by fluorescence microscopy. It was found that membrane associated Ca2+ decreased by the dihydropyridine nifedipine, whereas this Ca2+ pool was unaffected by the cellular polarization state, which was in contrast to cytosolic free Ca2+ as measured by fura-2. The results show that the odontoblast plasma membrane-associated Ca2+-pool can be modulated by nifedipine, thus being dependent on the conformational state of the L-type Ca2+ channels. PMID- 9733995 TI - Affinity chromatography purification of mitochondrial inner membrane proteins with calcium transport activity. AB - Immobilized calcium affinity chromatography was used to obtain a preparation enriched in calcium transporters from Triton X-100 extracts of rat liver mitochondria inner membranes (PPCT). The PPCT were reconstituted into preformed asolectin liposomes which contained 120 mM KCl as internal high K+ medium. 45Ca2+ uptake into proteoliposomes was studied under conditions favoring electrophoretic uptake, and H+i/45Ca2+o or Na+i/45Ca2+o exchange, to test for the presence of the three calcium transport modes present in mitochondria. 45Ca2+ uptake in liposomes was studied in parallel. Na+i/45Ca2+o exchange activity was not detectable. H+i/45Ca2+o exchange activity measured in the presence of a pH gradient (acid inside) obtained after suspension in low K medium in the presence of nigericin, was 100-200 nmoles 45Ca2+ per mg protein in 30 s. 45Ca2+ uptake in voltage dependent assays (a K+ diffusion membrane potential induced by valinomycin in the presence of methylamine) was not electrophoretic since it was stimulated by carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP) and probably due to secondary Ca2+/H+ countertransport. H+i/45Ca2+o uptake showed a saturable component at around 80 microM Ca and was coupled to an increase in internal pH in pyranine-loaded PPCT proteoliposomes. 45Ca2+ uptake in PPCT proteoliposomes could also be driven by a pH gradient obtained by raising external pH in high K+ medium. The results are consistent with the presence of a functional nH+/Ca2+ antiporter. Polyclonal antibodies raised against the PPCT were able to immunoprecipitate the H+/45Ca2+ uptake activity and recognized two major bands in the PPCT with molecular masses of about 66 kDa and 55 kDa. This is the first report of a partial purified protein(s) which may represent the H+/Ca2+ exchanger of the inner mitochondrial membrane, and represents an important step towards its identification. PMID- 9733996 TI - Protein structure and dynamics at high pressure. AB - The effect of pressure on the structure and dynamics of proteins is discussed in the framework of the pressure-temperature stability phase diagram. The elastic (reversible) properties, thermal expansion, compressibility and heat capacity, are correlated with the entropy, volume, and the coupling between entropy and volume fluctuations respectively. The experimental approaches that can be used to measure these quantities are reviewed. The plastic (conformational) changes reflect the changes in these properties in the cold, pressure and heat denaturation. PMID- 9733997 TI - Functional expression of a Ca2+-activated K+ channel in Xenopus oocytes injected with RNAs from the rat testis. AB - The present study investigated the feasibility of using Xenopus oocytes to express sperm ion channel by injection of RNAs extracted from the rat testis. The RNA-injected oocytes expressed an outwardly rectifying current which was dependent on K+ concentration and inhibitable by K+ channel blockers, charybdotoxin (CTX) and tetraethylammonium (TEA). The Ca2+ ionophore, ionomycin, could also stimulate current activation with similar current characteristics in the RNA-injected oocytes, suggesting the expression of a Ca2+-activated K+ channel. Immunolocalization indicated predominant Ca2+-activated K+ channel immunoreactivity associated with spermatogenic cells. Reverse transcriptase polymerase chain reaction studies confirmed the expression of the Ca2+-activated K+ channel mRNA in isolated spermatogenic cells. Our results suggest that ion channels and/or receptors of spermatogenic cells could be investigated using the Xenopus oocyte as an expression system. The present study also suggests that sperm may possess a Ca2+-activated K+ channel which has been implicated in the process of sperm activation and gamete interaction. PMID- 9733998 TI - Differential management of Ca2+ oscillations by anterior pituitary cells: a comparative overview. AB - Most electrical and ionic properties of anterior pituitary cells are common to all pituitary cell types; only gonadotropes exhibit a few cell specific features. Under basal conditions, the majority of pituitary cells in vitro, irrespective of their cell type, display spontaneous action potentials and [Ca2+]i transients that result from rhythmic Ca2+ entry through L-type Ca2+ channels. The main function of these action potentials is to maintain cells in a readily activable responsive state. We propose to call this state a 'pacemaker mode', since it persists in the absence of extrinsic stimulation. When challenged by hypothalamic releasing hormones, cells exhibit two distinct response patterns: amplification of pacemaker activity or shift to internal Ca2+ release mode. In the internal Ca2+ release mode, [Ca2+]i oscillations are not initiated by entry of external Ca2+, but by release of Ca2+ from intracellular stores. In somatotropes and corticotropes, GHRH or CRH triggers the pacemaker mode in silent cells and amplifies it in spontaneously active cells. In contrast, in gonadotropes GnRH activates the internal Ca2+ release mode in silent cells and switches already active cells from the pacemaker to the internal Ca2+ release mode. Interestingly, homologous normal and tumoral cells display the same type of activity in vitro, in the absence or presence of hypothalamic hormones. Pacemaker and internal Ca2+ release modes are likely to serve different purposes. Pacemaker activity allows long-lasting sequences of [Ca2+]i oscillations (and thus sustained periods of secretion) that stop under the influence of hypothalamic inhibitory peptides. In contrast, the time during which cells can maintain internal Ca2+ release mode depends upon the importance of intracellular Ca2+ stores. This mode is thus more adapted to trigger secretory peaks of large amplitude and short duration. On the basis of these observations, theoretical models of pituitary cell activity can be proposed. PMID- 9733999 TI - Cellular composition of the adult rat anterior pituitary is influenced by the neonatal sex steroid environment. AB - Growth hormone (GH) and prolactin (PRL) secretion differ significantly between adult males and females and this is due, at least in part, to the postpubertal hormone environment which affects GH and PRL gene expression, as well as somatotrope and lactotrope proliferation. However, the role of the neonatal steroid environment in this phenomenon is less well understood. We have used in situ hybridization to determine the number of GH and PRL mRNA containing cells, as well as the level of expression of these two hormones and of the pituitary transcription factor 1 (Pit-1). Neonatally castrated male rats that had been exposed to testosterone during the neonatal period, adulthood or during both periods, males castrated as adults, normal adult males and normal proestrous females were used. Orchidectomy of adult rats had no effect on the number of somatotropes or lactotropes, but significantly reduced GH and PRL mRNA levels. Neonatal castration significantly reduced the percentage of somatotropes and increased that of lactotropes in the adult male. In addition, GH and Pit-1 mRNA levels were reduced significantly, but PRL mRNA levels were not modified. Treatment of neonatally castrated males with testosterone during the neonatal period significantly increased the percentage of somatotropes and decreased the percentage of lactotropes compared to vehicle-treated animals. It also increased GH and Pit-1 mRNA levels, but did not affect PRL mRNA levels. Adult testosterone treatment significantly increased the percentage of both somatotropes and lactotropes, as well as GH, PRL and Pit-1 mRNA levels. Treatment of neonatally castrated males with testosterone during both the neonatal and adult periods returned the percentage of somatotropes and lactotropes, as well as GH, PRL and Pit-1 mRNA levels, to that of the intact male. These results suggest that, although the postpubertal steroid environment is important in determining anterior pituitary hormone synthesis and cellular composition, the neonatal steroid environment also plays an important role in this phenomenon. PMID- 9734000 TI - D2 dopamine-receptor-mediated inhibition of proliferation of rat lactotropes in culture is accompanied by changes in cell shape. AB - Dopaminergic agonists are effective in vivo in inhibiting lactotrope proliferation and prolactin (PRL)-secreting pituitary tumors. The purpose of the present study was to demonstrate in vitro actions of dopaminergic agents on proliferation and cell shape of rat lactotropes. Anterior pituitary cells cultured with serum-free, chemically defined medium were treated with dopaminergic agents and were labeled with 5-bromo-2'-deoxyuridine (BrdU) for 3 h before the end of culture. BrdU-labeling indices indicative of the proliferation rate of lactotropes were determined by double immunofluorescence staining for BrdU and PRL. Treatment with dopamine for 21 h decreased BrdU-labeling indices of lactotropes in a dose-dependent manner with a nadir at 3 x 10(-7) M. The inhibitory action of 10(-5) M dopamine appeared 15 h after the initiation of treatment and became pronounced with time up to 33 h. The dopamine action was mimicked by treatment with the D2 receptor agonist bromocriptine at concentrations over 10(-9) M. Phase-contrast microscopy revealed that the flat polygonal cell shape of cultured lactotropes had changed to a round refractive cell shape after treatment with dopamine or bromocriptine, and that these changes in cell shape exactly paralleled those in the BrdU-labeling index. The changes in cell shape of lactotropes were accompanied by changes in subcellular distribution of actin filaments. Pretreatment with 10(-7) M eticlopride, a D2 receptor antagonist, blocked the dopamine- or bromocriptine-induced changes in both BrdU labeling index and cell shape. These results suggest that (1) the in vitro experimental system established in the present study is a good model for studying the mechanism of the antiproliferative action of dopamine and (2) D2-receptor mediated inhibition of proliferation of lactotropes in serum-free culture is closely related to changes in actin organization and cell shape. PMID- 9734001 TI - Ovarian steroids modulate responsiveness to dopamine and expression of G-proteins in lactotropes. AB - The effects of chronic ovarian steroid treatment on the secretory activity of individual lactotropes and the mechanisms modulating their responsiveness to dopamine (DA) were studied. Female rats were ovariectomized (OVX) and implanted with Silastic capsules containing progesterone (P4), 17beta), 17beta-estradiol (E2) or both E2 (E2+P4). Ten days after surgery, anterior pituitaries were enzymatically dispersed and the reverse hemolytic plaque assay (RHPA) was performed to assess the release of prolactin (PRL) from individual lactotropes. RHPA was combined with immunocytochemistry (ICC) for PRL, Galphas or Gialpha3/Galphao proteins. E2 treatment alone or in combination with P4 increased the percentage of immunoreactive lactotropes among anterior pituitary cells. Incidence of active (plaque-forming) lactotropes, however, was increased both in P4-, and E2-treated rats and E2+P4 treatment increased it even further. While P4 treatment did not affect the frequency distribution of lactotropes, both E2 and E2+P4 treatments increased the large plaque-forming lactotrope population. This increase was reflected by the significantly greater mean plaque areas of lactotropes from E2- and E2+P4-treated rats compared to OVX or P4-treated animals. The responsiveness of lactotropes to DA from P4-treated rats did not differ from that of OVX rats: thus challenge with 1 microM DA inhibited the release of PRL, while 100 pM DA had no effect. E2 and E2+P4 treatments, however, profoundly changed the lactotrope's responsiveness: challenge with 1 microM DA had no effect and 100 pM DA resulted in moderate stimulation of PRL release in E2+P4 rats. Double-label ICC revealed that ovarian steroid treatments did not affect the expression of Galphas in lactotropes. The incidence of Gialpha3/Galphao-immunoreactive lactotropes, however, decreased after E2 treatment, alone or in combination with P4. Although expression of Galphas was similar in all plaque-forming cells regardless of plaque size, lactotropes expressing Gialpha3/Galphao were more likely to form small plaques in all treatment groups. These data suggest: (1) ovarian steroid treatment recruits quiescent lactotropes to release PRL; (2) E2 treatment alone or in combination with P4 increases the amount of PRL rleased by individual lactotropes; (3) E2 induced alterations in the frequency distribution and lactotrope responsiveness to DA may be due in part to a decreased expression of Gialpha3/Galphao. PMID- 9734002 TI - Nitric oxide (NO) stimulates gonadotropin secretion in vitro through a calcium dependent, cGMP-independent mechanism. AB - In the last few years, nitric oxide (NO) has emerged as an important intra- and intracellular messenger involved in the control of hypothalamic-pituitary function. The present experiments were undertaken in order to evaluate the pituitary component in the modulatory action of NO on gonadotropin secretion, as well as the second messenger pathway(s) involved. In a first step, we assessed LH and FSH secretion by hemipituitaries incubated in the presence of increasing concentrations of sodium nitroprusside (SNP), a potent NO donor, and cyclic guanosin monophosphate (cGMP), the second messenger for a wide range of NO actions. In addition, given that SNP induces the release of NO and cyanide ions, the response to SNP was tested in the presence of hemoglobin (an NO scavenger) or rhodanese + sodium thiosulfate (inactivators of cyanides) in order to ensure that the effects of SNP on gonadotropin secretion were mediated by the release of NO. SNP (10(-4)-10(-3) M) stimulated gonadotropin secretion in our incubation system, whereas cGMP, at all doses tested, was ineffective. Similar results were obtained using dispersed pituitary cells. The stimulatory action of SNP is attributable to its ability to induce NO release since it was blocked by hemoglobin, but preserved after incubation with rhodanese + sodium thiosulfate. In further experiments, we aimed to identify the mechanism(s) underlying SNP-induced gonadotropin secretion. First, to evaluate the involvement of calcium (Ca2+), the effects of SNP were analyzed in a calcium-free medium, after depletion of Ca2+ stores by caffeine, in the presence of the Ca2+ chelator ethylene glycol bis (p aminoethyl ether) N,N-tetra-acetic acid (EGTA), and after incubation with the Ca2+ channel blockers verapamil and nifedipine. Second, to confirm that cGMP is not involved in the stimulatory action of SNP, the effects of the latter on gonadotropin secretion were tested in the presence of the antagonists of the guanylyl cyclases oxadiazoloquinoxaline and LY 83,583. Our results showed that the stimulatory action of SNP on gonadotropin release is blunted in Ca2+-free medium and after incubation with EGTA, verapamil, nifedipine, and caffeine. On the contrary, the effect of SNP remained unaltered after antagonization of guanylyl cyclases. We conclude that NO, acting at the pituitary level, stimulates gonadotropin secretion through a calcium-dependent, cGMP-independent mechanism. PMID- 9734003 TI - Interaction between leptin and neuropeptide Y on in vivo growth hormone secretion. AB - Leptin, the product of the ob gene, is a recently discovered hormone secreted by adipocytes that regulates food intake and energy expenditure. Leptin has recently been shown to play a stimulatory role on GH secretion. The aim of the present study was to investigate whether leptin regulation of GH secretion was mediated by hypothalamic neuropeptide Y (NPY). We assessed the effect of leptin administration (10 microg, i.c.v.) and/or NPY (4 microg, i.c. v.) on fasted rats. Furthermore we administered leptin antiserum (10 microl, i.c.v.), anti-NPY serum (5 microl, i.c.v.) or normal rabbit serum (10 microl, i.c.v.) to freely moving fed rats. Spontaneous GH secretion was assessed over 6 h with blood samples taken every 15 min. Fed rats treated with anti-NPY serum exhibited a normal ultradian GH rhythm. However, administration of anti-NPY serum (5 microl, i.c.v., at 120 min) completely reversed the suppression induced by antileptin serum (10 microl, i.c.v., at 0 min) on plasma GH levels (area under the curve, AUC, 168 +/- 72 vs. 1,287 +/- 430 ng/ml/6 h; p < 0.01). In fasted rats, following NPY administration, GH levels remained suppressed throughout the 6 h studied. Besides, NPY administration completely blunted leptin-induced GH secretion as assessed by the AUC (28.5 +/- 11 vs. 520 +/- 220 ng/ml/6 h; p < 0. 01). Thus, it is possible that NPY mediates the effects of leptin on GH secretion. Alternatively, leptin and NPY could act through parallel pathways to alter GH release with NPY overcoming the stimulatory effect exerted by leptin on plasma GH levels. PMID- 9734004 TI - Cytokine-mediated growth hormone release from cultured ovine pituitary cells. AB - Previous studies have demonstrated that intravenous lipopolysaccharide (LPS) will increase concentrations of growth hormone (GH). One possible explanation for this may reside in the response of the pituitary to specific cytokines. This study sought to determine the effects of recombinant bovine tumor necrosis factor alpha (TNF), recombinant ovine (ro) interleukin-1alpha (IL-1alpha), roIL-1beta, ro interleukin-2 (IL-2), and ro gamma-interferon (INT) on GH release from cultured sheep pituitary cells. Sheep were sacrificed and pituitary cells cultured in DMEM with 10% fetal bovine serum for 3 days. On day 4, cells were washed and serum free DMEM added to cells. IL-1alpha and IL-1beta were used at 0.2, 2 and 20 ng/ml and the remaining cytokines at 2, 20 and 200 ng/ml. Neither IL-2 nor INT had effects on basal or on GH-releasing hormone (GRH)-stimulated GH release. TNF inhibited GRH-stimulated GH release (p < 0.05). Both IL-1alpha and IL-1beta stimulated GH release from cultured pituitary cells at all doses tested (p < 0.01). Neither IL-1alpha nor IL-1beta had an effect on GRH-stimulated GH release. IL-1 effects were inhibited by H-89 (p < 0.05; a protein kinase A inhibitor) and by nifedipine (p < 0.05; a calcium channel blocker). Both of these mechanisms are central signal transduction mechanisms mediating GRH-stimulated GH release. IL-1 stimulated GH release is partially inhibited (p < 0.05) by lipoxygenase pathway blockers. Phorbol myristate acetate downregulation of protein kinase C did not alter IL-1-stimulated GH release. IL-1beta increased the content of both GH and GH mRNA in cultured sheep pituitary cells. We conclude that IL-1 produces a strong stimulus to GH release, which is mediated by calcium entry and protein kinase A activation. IL-1 also activates lipoxygenase pathways. This latter pathway as well as calcium entry were shown to mediate LPS stimulation of GH release from cultured pituitary cells. The similarity between IL-1 and LPS signal transduction suggests that LPS may activate pituitary production of IL-1 to produce the stimulus to GH. The lack of inhibitory effects of INT, TNF and IL-2 as opposed to what is seen in the rat may suggest a partial mechanism to explain the different effects of LPS on GH release between sheep and that seen in cattle and rats. PMID- 9734005 TI - Growth hormone releasing hormone expression during postnatal development in growth hormone-deficient Ames dwarf mice: mRNA in situ hybridization. AB - Several genetic mutations in mice and rats that produce lifelong growth hormone (GH) deficiency result in overexpression of GH-releasing hormone (GHRH) mRNA in hypothalamic arcuate nucleus neurons. In order to examine the development of this condition, GHRH mRNA expression was quantified in Ames dwarf (df/df) and normal (DF/?) mice at 1 (day of birth), 3, 7, 14, 21 and 60 postnatal days (d) following in situ hybridization. Total mRNA was assessed using computer-assisted densitometry after X-ray film autoradiography, and mRNA expression per neuron was quantified by counts of grains per cell after emulsion autoradiography. Total GHRH mRNA was the same in dwarf and normal mice at 1, 3 and 7d. GHRH mRNA in dwarfs increased at 14d to 240% of that in DF/? (p < 0.005); the percentage overexpression in dwarf mice remained >/=200% through 60d, although total GHRH mRNA increased in both dwarfs and normals during this period. GHRH mRNA per neuron was the same in normal and dwarf mice at 1d, then increased in dwarfs to 190% of that in normals at 3d (p < 0.05), and rose to 300% of normal levels by 7d and beyond (p < 0. 005). There was no sexual dimorphism in expression by either measure in normal or dwarf mice. These results indicate that an increase in GHRH mRNA in Ames dwarf mice is first detectable at 3d, a period of approximately 7d after the failure to initiate GH production, which occurs normally at embryonic day 17.5. The onset of GHRH overexpression occurs earlier than the decline of either hypophysiotropic somatostatin or dopamine in Ames dwarf mice. This difference may be due to the stimulatory action of GHRH, as opposed to the inhibitory effects of factors examined previously. PMID- 9734006 TI - Sexual dimorphism in growth as measured by microknemometry: different responses to GH deficiency and exogenous GH administration. AB - To monitor growth, a novel noninvasive leg length measurement technique, called microknemometry, which allows daily observation of tibial growth rate, was used. The rat exhibits a striking sex-related difference in postpubertal growth. Exogenous GH administration results in a sexually dimorphic response, affecting growth in normal young female rats but not in males. Here we investigated how chronic GH deficiency affects male and female rat growth patterns. The degree of growth rate recovery was investigated after exogenous GH administration to chronically deficient males and females. The deficiency was induced by neonatal monosodium glutamate (MSG) treatment. Since the neonatal gonadal environment plays an important role in the dimorphic growth pattern, neonatal androgenization of female rats with testosterone or neonatal feminization of male rats by castration was performed and the growth pattern monitored. MSG treatment decreased pituitary GH content and plasma IGF I levels in both sexes, but caused a less marked reduction of female rat tibial growth and body weight gain than in males. Additionally, only MSG-treated males showed decreased pituitary LH content, so that the dimorphic action of MSG on the gonadal axis may contribute to the observed differences in growth rate. GH administration was able to increase leg length in all MSG-treated rats but was more effective in females, despite a similar restoration of plasma IGF I levels in both sexes. Although neonatal castration of male rats resulted in a reduction of tibial growth rate and body weight, and neonatal testosterone administration to female rats caused a slight increase in body weight, a complete modification of the gender-dependent growth pattern was not achieved, indicating that appropriate steroid environment is also needed in puberty and adulthood. PMID- 9734017 TI - Phylogenetic analysis and intrageneric structure of the genus Hyphomicrobium and the related genus Filomicrobium. AB - Almost complete 16S rDNA sequences from the type strains of seven species of the genus Hyphomicrobium and of Filomicrobium fusiforme have been determined. The Hyphomicrobium species from two phylogenetic clusters that are only moderately related to each other. While cluster I contains the type species Hyphomicrobium vulgare, Hyphomicrobium aestuarii, Hyphomicrobium hollandicum and Hyphomicrobium zavarzinii, cluster II comprises Hyphomicrobium facilis, Hyphomicrobium denitrificans and Hyphomicrobium methylovorum. Within the two species cluster, the species are highly related. Phylogenetically, Filomicrobium fusiforme clusters moderately with Hyphomicrobium species. The lack of distinguishing phenotypical properties presently excludes the possibility of describing cluster II as a new genus. PMID- 9734007 TI - Effects of amylin and salmon calcitonin on beta-endorphin-induced growth hormone and prolactin secretion in the rat. AB - In this study we examined the possible interplay of amylin (AMY) and salmon calcitonin (sCT) in the central control of growth hormone (GH) and prolactin (PRL) secretion in male rats. For this purpose we first compared effects of central intracerebroventricular (i.c.v.) admininstration of various doses of AMY (2.5-2,500 ng/rat) and sCT (2.2-220 ng/rat) on beta-endorphin (beta-END, 0.5 microg/rat)-induced GH and PRL secretion. AMY and sCT dose-dependently inhibited beta-END-induced GH secretion, whereas only sCT was able to inhibit beta-END induced PRL secretion. To examine whether the GH inhibitory effect of AMY was due to the possible cross-reactivity of AMY and sCT on the same receptors in the CNS, we pretreated some rats with the AMY antagonist (AMY8-37, 2. 5 microg/rat, i.c.v.). AMY8-37 significantly enhanced the GH-stimulatory action of beta-END. AMY8-37, administered prior to AMY and sCT, significantly removed the inhibitory effect of both AMY and sCT on beta-END-induced GH release, suggesting that both peptides mediate their response on GH through a common receptor. In vitro competition binding studies on rat hypothalamic membranes have shown that both AMY and sCT compete with [125I]rAMY binding with half inhibition (IC50) values of 3.6 x 10(-11) and 1.6 x 10(-10) M, respectively. Binding of [125I]sCT was inhibited by sCT with an IC50 of 1.09 x 10(-10) M and to a lesser extent by AMY with an IC50 of 1. 3 x 10(-6) M. Thus it is possible that the two peptides recognize a common hypothalamic receptor but with different affinities (sCT > AMY). Overall these data indicate that AMY behaves as a mimic of sCT in the central control of GH secretion. The failure of AMY, at variance with sCT, to modify the PRL-releasing activity of beta-END indicates that different receptor subtypes for sCT are involved in the endocrine effects of sCT and only those mediating the modulatory action of GH respond to AMY. PMID- 9734019 TI - Staphylococcus condimenti sp. nov., from soy sauce mash, and Staphylococcus carnosus (Schleifer and Fischer 1982) subsp. utilis subsp. nov. AB - Based on the sequence data of 23S rRNA of Staphylococcus carnosus, Staphylococcus piscifermentans, Staphylococcus aureus and Staphylococcus epidermidis, species specific probes were constructed. Their application revealed a heterogeneity within 18 strains previously identified as S. carnosus. Strains of this group were selected, and their 23S rRNA sequence was determined. It was revealed that the strains of S. carnosus can be placed in at least three sub-groups. This grouping was supported by physiological data and DNA-DNA similarity studies. Based on these results, were propose the new species Staphylococcus condimenti sp. nov. The type strain is S. condimenti F-2T (=DSM 11674T). The phylogenetic position of the new species within the radiation of other staphylococcal strains is reflected by a 16S nRNA-based tree. Furthermore, it is proposed to designate the new subspecies of Staphylococcus carnosus Schleifer and Fischer 1982, Staphylococcus carnosus subsp. utilis subsp. nov. The type strain of S. carnosus subsp. utilis is SK 11T (= DSM 11676T). PMID- 9734018 TI - Genetic analyses of the genus Nocardioides and related taxa based on 16S-23S rDNA internally transcribed spacer sequences. AB - The 16S-23S internally transcribed spacer (ITS) sequences were analysed to clarify inter- and intraspecific relationships among strains of the genus Nocardioides and the relationship between two Aeromicrobium species. The 16S-23S ITS regions from 33 Nocardioides strains, two Aeromicrobium species and Terrabacter tumescens were sequenced directly after polymerase chain reaction (PCR0 amplification and gamma exonuclease treatment. The genomes of some Nocardioides strains included two types of 16S-23S ITS sequences. The sizes of the 16S-23S ITS sequences of Nocardioides strains ranged from 328 to 539 bp. The 16S-23S ITS sequences of Aeromicrobium erythreum NSP37T, Aeromicrobium fastidiosum NSP38T and T. tumescens NSP39T were 349, 355 and 386 bp long, respectively. Nucleotide similarity among 16S-23S ITS sequences of Nocardioides albus strains and of Nocardioides simplex strains was 84.1-100% and 97.7-100%, respectively. The 16S-23S ITS sequence of Nocardioides luteus was identical to that of 'Nocardioides fulvus' NSP32T and was only 1 bp different from that of 'Nocardioides flavus' strains. However, the 16S-23S ITS sequences of 'N. fulvus' NSP33 showed only a low degree of similarity to 'N. fulvus' NSP32T (54.8%). The degree of 16S-23S ITS similarity between N. luteus NSP20T and N. ablus strains ranged from 85 to 93%. The mean nucleotide similarity values between the type strains of validly described Nocardioides species were highly divergent at 68:1 +/- 16.8%. The two Aeromicrobium species showed a level of 16S-23S ITS similarity of 71.2%. In this study, 16S-23S ITS sequences of the members of the genera Nocardioides and Aeromicrobium were useful for inferring the relationships between closely related strains and species. However, they were not found to be appropriate for elucidating the phylogenetic relationships between distantly related organisms at the genus level. PMID- 9734020 TI - Population genetic analysis of Serpulina pilosicoli and its molecular epidemiology in villages in the eastern Highlands of Papua New Guinea. AB - The population genetics of Serpulina pilosicoli and its molecular epidemiology in villages in the Eastern Highlands province of Papua New Guinea were investigated. Multilocus enzyme electrophoresis (MLEE) was used to analyse 164 isolates from humans and animals. These were divided into 33 electrophoretic types (ETs), four of which contained 65% of the isolates. The mean genetic diversity (n = number of ETs) for 145 human isolates was 0.18, and the mean number of alleles at five polymorphic loci was 2.6. The species appeared to be recombinant, as there was a lack of linkage disequilibrium, and 25% of all the possible combinations of alleles was present in the population. PFGE analysis using the enzymes M/ul and Sa/l divided 157 of the isolates into 99 PFGE types, demonstrating the existence of considerable strain diversity in a geographically restricted area. The two techniques were in excellent agreement; however, PFGE was more discriminatory for strain typing than was MLEE. Nine out of 19 (47.4%) culture-positive individuals were colonized by the same PFGE type of S. pilosicoli when retested after 6 weeks. For three individuals, the PFGE profiles of the second isolate differed from the first in only one or two DNA bands, while the other seven individuals were colonized with distinct PFGE types on each occasion. In two cases, strains with the same PFGE pattern were isolated from humans and dogs, suggesting that cross-species transmission of S. pilosicoli may occur naturally and that the infection can be zoonotic. PMID- 9734021 TI - Serpulina alvinipulli sp. nov., a new Serpulina species that is enteropathogenic for chickens. AB - Strain C1T is an anaerobic spirochaete that causes intestinal disease in chickens. Multilocus enzyme electrophoresis analysis and 16S rRNA sequence comparisons have indicated that this spirochaete is a Serpulina strain. In these investigations, various phenotypic and genomic properties useful for establishing a taxonomic identity for strain C1T were studied. As determined by electron microscopy, cells of the spirochaete measured 8-11 x 0.22-0.34 mum and had a typical spirochaete ultrastructure. Each cell had 22-30 flagella. C1T cells formed weakly beta-haemolytic colonies on trypticase soy agar plates containing 5% bovine blood. The spirochaete reached maximum population densities of 10(9) cells ml-1 with a 2-4 h population doubling time in brain heart infusion broth containing 10% calf serum (BHIS broth). C1T cultures in BHIS broth were positive in tests for hippurate hydrolysis and negative for indole production. Glucosamine, N-acetyglucosamine, glucose, fructose, maltose and mannose were growth substrates for the spirochaete in heart infusion broth containing 7% calf serum (HS broth). During growth in HS broth beneath an O2/N2 (1:99) atmosphere, cells of the spirochaete consumed O2 and glucose and produced H2, CO2, acetate, butyrate and ethanol. Strain C1T DNA had a G+C content of 24.6 mol%. Based on DNA DNA hybridization analyses, the DNA of strain C1T exhibited 24-39% relative reassociation with DNA of Serpulina hyodysenteriae, Serpulina innocens, Serpulina pilosicoli, Serpulina murdochii and Serpulina intermedia. These results indicate that chicken spirochaete strain C1T has many phenotypic properties common to Serpulina species and, based on DNA hybridization analysis, represents a unique Serpulina species. For this new species the name Serpulina alvinipulli is proposed, for which the type strain is C1T (= ATCC 51933T). PMID- 9734022 TI - Carnimonas nigrificans gen. nov., sp. nov., a bacterial causative agent for black spot formation on cured meat products. AB - Nine different strains, CTCBS1T or CTCBS9, were identified to be the causative agents of black spots on the surface of raw cured meat products. The formation of black spots under aerobic conditions is reproducible upon reinoculation of meat products with any of these strains, indicating that they are the causative agent. The strains were Gram-negative, catalase-positive and obligately aerobic rods. The G+C content of DNA of strain CTCBS1T is 56.0 +/- 0.3 mol%. The content of non polar main fatty acids were 16:0, 16:1, 18:1 and 19:0 cyc. Its phylogenetic position was elucidated by comparative sequence analysis of the 16S rRNA gene. Overall sequence similarity to other bacteria does not exceed 93.3%. Isolate CTCBS1T clustered phylogenetically within the gamma-subclass of the Proteobacteria and is closely related to members of Halomonas (90:5-91.9%) and to Zymobacter palmae (93.3%). A genetic homogeneity of the nine strains was demonstrated by M13 random amplified polymorphic DNA-PCR, whereas differentiation from other genera, e.g. Zymobacter and Pseudomonas, could easily be achieved by their chemotaxonomic characteristics. Taxonomic data revealed the status of a separate genus for which the name Carnimonas gen. nov., sp., nov. is proposed. Despite chemotaxonomic and physiological similarities, the new genus is at present not a member of the family Halomonadacease because of the lack of two out of 15 descriptive 16S rRNA signature sequences. The first member of the new genus is Carnimonas nigrificans. The use of a specific, 16S rRNA-targeted oligonucleotide primer allowed the identification of all nine strains of C. nigrificans in a PCR assay. Toxicological studies showed no pathogenic potential for C. nigrificans strain CTCBS1T (CECT 4437T). PMID- 9734023 TI - Rhizobium huautlense sp. nov., a symbiont of Sesbania herbacea that has a close phylogenetic relationship with Rhizobium galegae. AB - The nitrogen-fixing rhizobial symbionts of Sesbania herbacea growing in the nature reserve at the Sierra de Huautla, Mexico, were isolated and characterized. All 104 isolates together with the type strain for Rhizobium galegae, HAMBI 540T, had similar 16S rRNA genes as revealed by PCR-RFLP analysis. Similarity in the sequences of the 16S rRNA genes placed the isolates on a phylogenetic branch shared with R. galegae. Among 66 randomly selected isolates, three closely related electrophoretic alloenzyme types (ETs) were identified, which were distinct from 10 ETs distinguished among 23 strains of R. galegae. A new species Rhizobium huautlense, represented by the Sesbania isolate SO2T, is proposed based upon low estimates of DNA relatedness between our chosen type strain and the type strains for the other species, the dissimilarity of the nucleotide sequence of the 16S rRNA genes, and their distinct ETs compared with R. galegae. The description of R. huautlense is significant because in the reconstruction of the phylogeny at R. huautlense there was a shift in the node of the branch of Agrobacterium vitis relative to that of R. galegae. The revised phylogenetic tree would tend to indicate common ancestry between R. galegae and Rhizobium leguminosarum. PMID- 9734024 TI - Desulfurobacterium thermolithotrophum gen. nov., sp. nov., a novel autotrophic, sulphur-reducing bacterium isolated from a deep-sea hydrothermal vent. AB - A thermophilic, anaerobic, strictly autotrophic, sulphur-reducing bacterium, designated BSAT (T = type strain), was isolated from a deep-sea hydrothermal chimney sample collected at the mid-Atlantic ridge. Gram-negative cells occurred singly or in pairs as small highly motile rods. Spores were not observed. The temperature range for growth was 40 to 75 degrees C, with an optimum at 70 degrees C. The pH range for growth at 70 degrees C was from 4.4 to 7.5, with an optimum around 6.0. The sea salt concentration range for growth was 15-70 gI(-1) with an optimum at 35 gI(-1). Elemental sulphur, thiosulphate and sulphite were reduced to hydrogen sulphide. Sulphate and cystine were not reduced. The G+C content of the genomic DNA was 35 mol%. Phylogenetic analyses of the 16S rRNA gene indicated that the strain was a member of the domain Bacteria and formed a branch that was almost equidistant from members of the orders Aquificales and Thermotogales. The new organism possesses phenotypic and phylogenetic traits that do not allow its classification as a member of any previously described genus; therefore, it is proposed that this isolate should be described as a member of a novel species of a new genus, Desulfurobacterium gen. nov., of which Desulfurobacterium thermolithotrophum sp. nov. is the type species. The type strain is BSAT (= DMS 11699T). PMID- 9734025 TI - Phylogenetic analysis of cultivable oral treponemes from the Smibert collection. AB - Dr. Robert Smibert from the Virginia Polytechnic Institute, USA, isolated and collected over 200 strains of oral treponemes over a 20-year period. Dr. Smibert, Dr. W.E.C. Moore and Dr. L.V. Moore separated these isolates and reference strains into different groups on the basis of cellular fatty acid analysis. In this study, the 16S rRNA genes were sequenced for 47 strains that were representative of these groups. Five distinct species were identified on the basis of 16S rRNA sequence comparisons; two of these species are newly named and three have not yet been characterized. The first species, designated Treponema Smibert-1, was represented by the single strain D4B-1 and was later identified as the newly described Treponema maltophilum. However, strain D4B-1 possessed a different flagellar arrangement to that of T. maltophilum. The second species, Treponema Smibert-2, was represented by nine isolates that possessed identical 16S rRNA gene sequences. The closest relatives of this species were Treponema Smibert-3 and Treponema Smibert-4 at approximately 90% sequence similarity. Within Treponema Smibert-2, there was no correlation between phylogenetic analysis and cellular fatty acid analysis since six different cellular fatty acid groups represented the nine strains. Treponema Smibert-3 (strain D36ER-1) and Treponema Smibert-4 (D62CR-12) were each represented by only a single strain and were closely related to each other at 98% sequence similarity. Strain D36ER-1 of Treponema Smibert-3 was identified as belonging to the not-yet-cultivated phylotype 20 [Choi, B.K., Paster, B.J., Dewhirst, F.E. & Gobel, U.B. (1994). Infect Immun 62, 1889-1895]. Strain D62CR-12 of Treponema Smibert-4 was nearly identical in sequence to the newly described Treponema amylovorum. The fifth species, Treponema Smibert-5, was represented by a single strain, D120CR-1, and was closely related at about 98% sequence similarity to the three subspecies of Treponema socranskii. The polygenetic analyses of strains of Treponema vincentii and of subspecies of T. socranskii are also reported. The closest oral relatives of T. vincentii were Treponema medium at 98.7% sequence similarity and Treponema denticola at 91.5% sequence similarity. T. socranskii subspp. socranskii, buccale and paredis formed three separate phylogenetic branches with sequence similarities of about 98% to each other. The closest relative of the subspecies of T. socranskii and of Smibert-5 was Smibert-2 at about 86% sequence similarity. Historic reference strains Fuji, 'Treponema ambigua', Fm, Ichelson-2, N-39, TD2, TRRD, MRB, IPP, Jethro and T32A, as well as an unkown strain designated only as Treponema oralis, were identified as strains of T. denticola. Reference strains Fuji, Jethro, T32A, and IPP plus three isolates of the Smibert collection were also contaminated with a mycoplasma as determined by 16S rRNA comparative analysis. Consequently, spirochaetal cultures should be screened for mycoplasmas. There are presently at least ten species of cultivable oral species of treponema with the cut-off for separate species designation at about 98% sequence similarity. However, DNA-DNA reassociation experiments are necessary to differentiate species when 16S rDNA sequence similarities are at about this level. PMID- 9734027 TI - Phylogenetic heterogeneity within the genus Herpetosiphon: transfer of the marine species Herpetosiphon cohaerens, Herpetosiphon nigricans and Herpetosiphon persicus to the genus Lewinella gen. nov. in the Flexibacter-Bacteroides Cytophaga phylum. AB - Analysis of the 16S rDNA sequences of species currently assigned to the genus Herpetosiphon revealed intrageneric phylogenetic heterogeneity. The thermotolerant freshwater species Herpetosiphon geysericola is most closely related to the type species Herpetosiphon aurantiacus in the Chloroflexus subdivision of the green non-sulfur bacteria. The marine species Herpetosiphon cohaerens, Herpetosiphon nigricans and Herpetosiphon persicus, on the other hand, were found to form a cluster with sheathed bacterium Haliscomenobacter hydrossis in the Saprospira group of the Flexibacter-Bacteroides-Cytophaga (FBC) phylum. A proposal is made to transfer these marine species to the genus Lewinella gen. nov. as Lewinella cohaerens comb. nov., Lewinella nigricans comb. nov. and Lewinella persica comb. nov. The marine sheathed gliding bacterium Flexithrix dorotheae was also found to be a member of the FBC phylum but on a separate phylogenetic line to the marine herpetosiphons now assigned to the genus Lewinella. PMID- 9734026 TI - Species identification of Legionella via intergenic 16S-23S ribosomal spacer PCR analysis. AB - Species identification of Legionella in routine laboratory testing is hampered by the lack of highly discriminatory phenotypic tests. Amplification polymorphism of the intergenic 16S-23S spacer regions (ISR) has been previously developed for identification of species within the Legionellaceae [Hookey, J.V., Birtles, R.J. & Saunders, N.A. (1995). J Clin Microbiol 33, 2377-2381], but it did not provide enough resolution to distinguish all members of the bluish-white autofluorescent species and the red autofluorescent group of the Legionellaceae. By choosing new primers that target regions 4 (positions 1521-1541 of Escherichia coli 16S rRNA gene) and 6 (positions 114-132 of E.coli 23S rRNA gene) within the rDNA operon close to the 16S-23S intergenic spacer, 34 profiles were determined among the 79 type and reference strains representing 42 species that were tested. Analysis of the RFLP generated after Hinfl restriction digestion of the PCR products further improved the method, allowing complete discrimination among the species and subspecies of Legionella tested. Twenty-three well-identified strains from unrelated origins belonging to seven species gave amplification patterns identical to that of their type strain. The technique was also tested on 80 field isolates that could not be unequivocally assigned to groups by phenotypic methods. Seventy-two per cent (58/80) of these isolates had a profile identical to that of a type strain, while 27% (22/80) may correspond to new taxa since their ISR-PCR profiles did not match any of the known profiles. PMID- 9734028 TI - Union of the genera Microbacterium Orla-Jensen and Aureobacterium Collins et al. in a redefined genus Microbacterium. AB - The 16S rRNA gene sequences of 19 strains, 11 strains representing validated Aureobacterium or Microbacterium species and eight strains of non-valid species or isolates, were determined. These sequences were aligned with the sequences of other validated Aureobacterium and Microbacterium species and related actinobacteria. A comparative sequence analysis of 43 strains revealed that the species of the genera Aureobacterium and Microbacterium form a monophyletic association in which species of both genera are intermixed. The high similarity in phylogenetic properties found in the species within both genera and the close relationship in physiological and chemotaxonomic features other than the diamino acid in the cell wall, provided strong evidence that the genera Aureobacterium and Microbacterium should be unified. An emended genus Microbacterium is proposed for the two combined genera. The following validated Aureobacterium species were combined to the genus Microbacterium: Aureobacterium arabinogalactanolyticum to Microbacterium arabinogalactanolyticum, Aureobacterium barkeri to Microbacterium barkeri, Aureobacterium esteraromaticum to Microbacterium esteraromaticum, Aureobacterium flavescens to Microbacterium flavescens, Aureobacterium keratanolyticum to Microbacterium liquefaciens, Aureobacterium luteolum to Microbacterium luteolum, Aureobacterium saperdae to Microbacterium saperdae, Aureobacterium schleiferi to Microbacterium schleiferi, Aureobacterium terrae to Microbacterium terrae, Aureobacterium terregens to Microbacterium terregens, Aureobacterium testaceum to Microbacterium testaceum, and Aureobacterium trichothecenolyticum to Microbacterium trichothecenolyticum. PMID- 9734029 TI - Evaluation of the relatedness of Brucella spp. and Ochrobactrum anthropi and description of Ochrobactrum intermedium sp. nov., a new species with a closer relationship to Brucella spp. AB - The relatedness of Brucella spp. and Ochrobactrum anthropi was studied by protein profiling, Western blot, immunoelectrophoresis and 16S rRNA analysis. Whole-cell and soluble proteins of brucellae and O. anthropi showed serological cross reactivities quantitatively and qualitatively more intense than those existing with similar extracts of Agrobacterium spp. Numerical analysis of Western blot profiles of whole-cell extracts showed that O. anthropi LMG 3301 was closer to Brucella spp. than to O. anthropi LMG 3331T, a result not obtained by protein profiling. These differences were not observed by Western blot with soluble fractions, and immunoelectrophoretic analyses suggested that this was due to destruction of conformational epitopes in Western blot procedures with the subsequent simplification of antigenic profile. Analysis of the 16S rRNA sequences of strains previously used in the species definition confirmed that strain LMG 3301, and also LMG 3306, were closer to the brucellae, and that LMG 3331T was in a separate cluster. The LMG 3301 and the LMG 3331T clusters could also be separated by their different colistin sensitivity and by PCR with 16S rRNA Brucella primers, and both methods showed strains of both clusters among clinical isolates classified as O. anthropi by conventional tests. These results and those of previous DNA-DNA hybridization studies [Holmes, B., Popoff, M., Kiredjian, M. & Kersters, K. (1988). Int J Syst Bacteriol 38, 406-416] show that the LMG 3301 cluster and related clinical isolates should be given a new species status for which the name Ochrobactrum intermedium sp. nov. is proposed (type strain is LMG 3301T=NCTC 12171T = CNS 2-75T). PMID- 9734030 TI - Pseudoalteromonas bacteriolytica sp. nov., a marine bacterium that is the causative agent of red spot disease of Laminaria japonica. AB - An aerobic, polarly flagellated marine bacterium that produces a prodigiosin-like pigment was isolated from the red-spotted culture beds of Laminaria japonica. Five isolates had unique bacteriolytic activity for both Gram-positive and negative bacteria, which had never been observed among Alteromonas or related species. The isolates were identified as the causative agent of red spot disease of L. japonica seeds. The phenotypic features of the isolates were similar to these of Pseudoalteromonas rubra ATCC 29570T, but they could be differentiated using 10 traits (growth at 37 degrees C, requirement for organic growth factors, bacteriolytic activity, utilization of sucrose, N-acetylglucosamine, fumarate, succinate, D-galactose, L-proline and acetate). The G+C content of DNAs from the isolates was 44-46 mol%. The isolates constitute a new species, distinct from the other Alteromonas and Pseudoalteromonas species, as shown by DNA-DNA hybridization experiments and phylogenetic clustering of 16S rRNA gene sequences, for which the name Pseudoalteromonas bacteriolytica sp. nov. (type strain = IAM 14595T) is proposed. A set of phenotypic features which differentiate this new species from closely related Pseudoalteromonas and Alteromonas species is provided. PMID- 9734031 TI - Microvirgula aerodenitrificans gen. nov., sp. nov., a new gram-negative bacterium exhibiting co-respiration of oxygen and nitrogen oxides up to oxygen-saturated conditions. AB - A denitrifier micro-organism was isolated from an upflow denitrifying filter inoculated with an activated sludge. The cells were Gram-negative, catalase- and oxidase-positive curved rods and very motile. They were aerobic as well as anoxic heterotrophs that had an atypical respiratory type of metabolism in which oxygen and nitrogen oxides were used simultaneously as terminal electron acceptors. The G&C content was 65 mol%. Our isolate was phenotypically similar to Comamonas testosteroni, according to classical systematic classification systems. However, a phylogenetic analysis based on the 165 rRNA sequence showed that the aerobic denitrifier could not be assigned to any currently recognized genus. For these reasons a new genus and species, Microvirgula aerodenitrificans gen. nov., sp. nov., is proposed, for which SGLY2T is the type strain. PMID- 9734032 TI - Selenomonas lipolytica sp. nov., an obligately anaerobic bacterium possessing lipolytic activity. AB - A novel, oligately anaerobic bacterium capable of hydrolysing lipids was isolated from a tropical anaerobic lagoon receiving waste water from an edible oil mill. The isolate had many characteristics similar to those of members of the genus Selenomonas. The isolate showed lipolytic activity on tributyrin, triolein and groundnut oil in qualitative plate clearance assays, which has not been reported for the type strain of the genus Selenomonas. It did not require n-valerate supplementation for growth on glucose. Acetate and propionate were the only volatile fatty acids produced from glucose fermentation with propionate as the major end product. The isolate could grow optimally at pH 6.8 and at a temperature of 40 degrees C. It could tolerate NaCl concentrations of up to 40 g l-1. The G&C content of the DNA was 40 mol% as determined by thermal denaturation analysis. Comparison of partial 165 rRNA gene sequences revealed that the isolate was most closely related to genus Selenomonas with 91% sequence similarity (250 bp compared) to Selenomonas ruminantium strain GA 192. On the basis of the results obtained in the present investigation, it is suggested that a new species of Selenomonas should be created for this novel isolate and the name Selenomonas lipolytica is proposed for this new species. The type strain is strain CF1BT (= MCMB 505T). PMID- 9734033 TI - Reclassification of species of the spiral-shaped phototrophic purple non-sulfur bacteria of the alpha-Proteobacteria: description of the new genera Phaeospirillum gen. nov., Rhodovibrio gen. nov., Rhodothalassium gen. nov. and Roseospira gen. nov. as well as transfer of Rhodospirillum fulvum to Phaeospirillum fulvum comb. nov., of Rhodospirillum molischianum to Phaeospirillum molischianum comb. nov., of Rhodospirillum salinarum to Rhodovibrio salexigens. AB - The 165 rDNA sequence of Rhodospirillum mediosalinum was determined and compared with corresponding sequences from other spiral-shaped purple non-sulfur bacteria classified as or related to the genus Rhodospirillum in the alpha subclass of the Proteobacteria. Sequence similarities separate the currently recognized Rhodospirillum species into five different groups with no more than 91% sequence similarity, clearly indicating the necessity to recognize these groups as different genera. Major diagnostic properties of these bacteria are compared and new genera Phaeospirillum gen. nov., Roseospira gen. nov., Rhodothalassium gen. nov. and Rhodovibrio gen. nov. are described with the species Phaeospirillum fulvum comb. nov., Phaeospirillum molischianum comb. nov., Rhodovibrio salinarum comb. nov., Rhodovibrio sodomensis comb. nov., Rhodothalassium salexigens comb. nov. and Roseospira mediosalina comb. nov. The genus Rhodospirillum is represented by Rhodospirillum rubrum and Rhodospirillum photometricum and an emended description of this genus is also given. PMID- 9734034 TI - Staphylococcus hominis subsp. novobiosepticus subsp. nov., a novel trehalose- and N-acetyl-D-glucosamine-negative, novobiocin- and multiple-antibiotic-resistant subspecies isolated from human blood cultures. AB - A new subspecies, Staphylococcus hominis subsp. novobiosepticus, isolated from human blood cultures, a wound, a breast abscess and a catheter tip, is described on the basis of a study of 26 strains isolated between 1989 and 1996. DNA-DNA reassociation reactions, conducted under stringent conditions, and macrorestriction pattern analysis demonstrated that these strains are closely related to previously characterized S. hominis strains isolated from human skin and clinical specimens, but are significantly divergent. S. hominis subsp. novobiosepticus can be distinguished from S. hominis (now named S. hominis subsp. hominis) by its combined characteristics of novobiocin resistance and failure to produce acid aerobically from D-trehalose and N-acetyl-D-glucosamine. Furthermore, all 26 strains of the new subspecies are resistant to nalidixic acid, penicillin G, oxacillin, kanamycin and streptomycin, and were either resistant or had intermediate resistance to methicillin and gentamicin. Most strains were also resistant to erythromycin, clindamycin, chloramphenicol, trimethoprim/sulfamethoxazole and ciprofloxacin. Based on a comparison of the sequences of a 1001 bp mecA amplification product from reference methicillin resistant staphylococci, the mecA gene present in S. hominis subsp. novobiosepticus was identified as homologue A, commonly found in S. aureus and many coagulase-negative staphylococcal species. The type strain of S. hominis subsp. novobiosepticus is ATCC 700236T. Descriptions of S. hominis subsp. novobiosepticus subsp. nov and S. hominis subsp. hominis are given and the description of S. hominis is emended. PMID- 9734035 TI - Phylogenetic relationships of Pseudomonas putida strains deduced from the nucleotide sequences of gyrB, rpoD and 16S rRNA genes. AB - Phylogenetic analysis of 20 Pseudomonas strains (Pseudomonas putida, Pseudomonas fluorescens and Pseudomonas chlororaphis) was conducted by using the nucleotide sequences of the genes for 16S RNA, DNA gyrase B subunit (gyrB) and RNA polymerase delta 70 factor (rpoD), which have been determined by the direct sequencing of PCR-amplified fragments. On the basis of gyrB and rpoD sequences, these strains were split into two major clusters: one including the type strain of P. putida and all biovar A strains and the other including all P. putida biovar B strains, P. fluorescens stains and the P. chlororaphis strain. In the phylogenetic tree reconstructed from the 16S rRNA sequences included variable regions, P. Putida biovar A and B strains were not separated into two independent clusters, whereas in the phylogenetic tree reconstructed from the 16S rRNA sequences excluding the variable region sequences, these strains were separated into P. putida biovar A and biovar B clusters. The pairwise distances estimated from the variable regions of 16S rRNA correlated poorly with the synonymous distances estimated from the gyrB and rpoD genes. On the other hand, a highly significant correlation was observed between the pairwise distances estimated from the non-variable regions of 16S rRNA and the synonymous distances from gyrB and rpoD genes. Consequently, only the 16S rRNA sequences in the non-variable regions should be used for the phylogenetic analysis. The gyrB and rpoD analyses showed the necessity for the reclassification of P. putida biovar B strains. PMID- 9734036 TI - Methanocalculus halotolerans gen. nov., sp. nov., isolated from an oil-producing well. AB - Two irregular coccoid methanogens designated SEBR 4845T and FR1T were isolated from an oilfield in Alsace, France. Strain SEBR 4845T (T = type strain) is a hydrogenotrophic halotolerant methanogen, which grows optimally at 5% NaCI (w/v) and tolerates up to 12% NaCI. It does not use methylated compounds and therefore cannot be ascribed to any of the known genera of the halophilic methylotrophic methanogens. It differs from hydrogenotrophic members of the orders Methanococcales and Methanomicrobia les in the NaCI growth range (0-12% NaCI), which is the widest reported to data for any hydrogenotrophic methanogen. 16S rRNA gene sequence analysis indicated that strain SEBR 4845T is a novel isolate for which a new genus is proposed, Methanocalculus halotolerans gen. nov., sp. nov. (= OCM470T) that might be indigenous to the oilfield ecosystem. Strain FR1T (=OCM 471) is a moderately halophilic methanogen which growths optimally at 10% NaCI and tolerates up to 20% NaCI. It grows on trimethylamine and methanol as carbon and energy sources. The G+C content of its DNA is 43 mol%. It is therefore phenotypically and genotypically related to members of the genus Methanohalophilus. This report provides evidence that methylotrophic and hydrogenotrophic, but not aceticlastic methanogens are present in a saline subsurface oilfield environment, as already observed in surface saline to hypersaline environments. PMID- 9734037 TI - Paenibacillus campinasensis sp. nov., a cyclodextrin-producing bacterium isolated in Brazil. AB - An alkaliphilic, endospore-forming bacterium isolated from Brazilian soil was taxonomically studied and is proposed as a new Paenibacillus species. This organism (strain 324T) was particularly distinguishable from other Paenibacillus species by its ability to grow optimally at pH 10 and 40 degrees C. The DNA G+C content was 5.0 mol%. The diamine acid of the cell-wall peptidoglycan was meso diaminopimelic acid. MK-7 was the predominant menaquinone and anteiso-C15:0 was the major fatty acid. Levels of 16S rDNA similarity between strain 324T and other Paenibacillus species were 90.6-95.9%. Phylogenetically, strain 324T formed an evolutionary lineage distinct from other species within the evolutionary radiation encompassing the genus Paenibacillus. Based on phenotyic and chemotaxonomic properties, and phylogenetic inference, it is proposed that strain 324T should be placed in the genus Paenibacillus as a new species is strain 324T should be placed in the genus Paenibacilus as a new species, Paenibacillus campinasensis. This type strain of the new species is strain 325T (= KCTC 0364BP). PMID- 9734038 TI - Phylogenetic analysis of spotted fever group rickettsiae by study of the outer surface protein rOmpA. AB - Rickettsiae are classified in the order Rickettsiales and have been included in the alpha subclass of the class Proteobacteria on the basis of 16S rRNA gene sequence comparison. To estimate the evolutionary forces that have shaped the members of the spotted fever group (SFG) rickettsiae, the ompA gene (apart from the tandem repeat units), encoding an antigenic high-molecular-mass membrane protein specific for the group, was amplified and sequenced from 21 isolates. The phylogenetic relationship between SFG rickettsiae were inferred from the comparison of both the gene and derived protein sequences, using the parsimony, neighbor-joining and maximum-likelihood methods. Three strongly supported phylogenetic sub-groups were distinguished: first, the Rickettsia conorii complex (R. conorii Malish, R. conorii M1, R. conorii Moroccan, R. conorii Indian tick typhus, Astrakhan fever rickettsia and Israeli tick typhus rickettsia); second, a cluster including Rickettsia africae, strain S, Rickettsia parkeri, Rickettsia sibirica and 'Rickettsia mongolotimonae'; and, third, a cluster including Rickettsia aeschlimannii, Rickettsia rhipicephali, Rickettsia massiliae, Bar 29 and Rickettsia montanensis. Rickettsia rickettsii, Rickettsia japonica, Rickettsia slovaca and Thai tick typhus rickettsia did not cluster with any other Rickettsia species. To test whether positive selection was responsible for sequences diversity, rates of synonymous and nonsynonymous nucleotide substitutions were compared for Rickettsia ompA alleles and indicated that this gene is undergoing neutral evolution. PMID- 9734039 TI - Leptospira fainei sp. nov., isolated from pigs in Australia. AB - Pathogenic leptospires can be causative agents of reproductive problems in pigs. Cultures of uteri and kidneys from two pigs herds in New South Wales and Victoria (Australia) yielded five strains identified as Leptospira on morphological and cultural grounds. Phenotypic characteristics (growth at 13 and 30 degrees C, growth in the presence of 8-azaguanine) were intermediate between those of pathogenic and saprophytic leptospires. No cross-agglutination was observed with reference antisera representing the 24 pathogenic serogroups and the main saprophytic ones. Antiserum against one of the strains did not agglutinate reference stains representative of any serogroup. This provided evidence of a new serovar, designated hurstbridge. Genomic characterization of the five strains was achieved using five molecular approaches. Mapped restriction site polymorphisms in the rrs (16S rRNA) gene were not related to those of any reference strains. Arbitrarily primed PCR fingerprints suggested clonality of the five strains. The strains all showed an identical and unique PFGE profile. PCR, using primers specific for the rrs gene of pathologic leptospires, amplified corresponding sequences from the strains. DNA-DNA hybridization (and reciprocal experiments) using the S1 nucleas/TCA method was performed between one of the strains and the reference strains of Leptospira species. The homology ranged from 0 to 36% (the latter being was Leptospira inadai) thus satisfying the criterion of a new species, Leptospira fainei (type strain BUT 6T). Phylogenetic analysis of 16S rRNA sequence showed that L. fainei and L. inadai formed a clade separate from the previously recognized 'saprophyte' and 'pathogen' clades. PMID- 9734040 TI - Delimiting the genus Staphylococcus through description of Macrococcus caseolyticus gen. nov., comb. nov. and Macrococcus equipercicus sp. nov., and Macrococcus bovicus sp. no. and Macrococcus carouselicus sp. nov. AB - Four species of the newly proposed genus Macrococcus, namely macrococcus caseolyticus gen. nov., comb. nov. (formerly Staphylococcus caseolyticus Schleifer, Kilpper-Balz, Fischer, Faller and Endl 1982, 19VP), Macrococcus equipercicus sp. nov., Macrococcus bovicus sp. nov. Macrococcus carouselicus sp. nov., are described on the basis of a phylogenetic analysis comparing 16S rRNA sequences, DNA-DNA liquid hybridization, DNA base composition, normalized ribotype patterns, macrorestriction pattern analysis and estimation of genome size using PFGE, cell wall composition, phenotypic characteristics and plasmid profiles. Compared with their closet relatives, members of the genus Staphylococcus, these organisms demonstrated significantly lower 16S rRNA sequence similarities (93.4-95.3%), higher DNA G+C content (38-45 mol%), absence of cell wall teichoic acids (with the possible exception of M. caseolyticus), unique ribotype pattern types and macrorestriction patterns, smaller genome size (approx. 1500-1800 kb) and generally larger Gram-stained cell size (1.1-2.5% microns in diameter). Macrococci can be distinguished from most species of staphylococci (except Staphylococcus sciuri, Staphylococcus vitulus and Staphylococcus lentus) by thier oxidase activity. The four Macrococcus species can be distinguished from one another on the basis of DNA-DNA hybridization, ribotype pattern types, macrorestriction patterns and their phenotypic properties, including colony morphology, cell morphology, haemolysins, Staphy Latex agglutination, acid production from a variety of carbohydrates, acetoin production, nitrate reduction, aesculin hydrolysis, and DNase and urease activities. The type species is M. equipercicus. The type strains of M. equipercicus, M. caseolyticus, M. bovicus and M. carouselicus are ATTCC 51831T (= DD 9350T) ATCC 13548T (= TDD 4508T) (Schleifer et al. 1982, ATCC 51825T (= DD 4516T) and ATCC 51828T (= DD 9348), respectively. PMID- 9734041 TI - Thermocladium modestius gen. nov., sp. nov., a new genus of rod-shaped, extremely thermophilic crenarchaeote. AB - Three strains of novel, extremely thermophilic, rod-shaped crenarchaeotes were isolated from acidic hot spring areas in Japan. Cells of the three stains were straight or slightly curved rods and occasionally branched out singly or extensively, or had spherical bodies protruding at the ends of the cells. They were heterotrophs that grew anaerobically or microaerobically. The presence of CO2 in the gas phase, archaeal cell-extracts and a vitamin mixture stimulated growth of the strains. Growth occurred at 45-82 degrees C and pH 2.6-5.9 and was optimal around 75 degrees C and pH 4.0. The strains utilized glycogen, starch, gelatin and various proteinaceous complex compounds as carbon sources. They required sulfur, thiosulfate or L-cystine as possible electron acceptors. The lipids mainly consisted of various cyclic glycerol-bisdiphytanyl-glycerol tetraethers. The G+C contents of the genomic DNAs were 52 mol%. Comparison of the 16S rDNA sequences indicated that they belonged to a separate lineage in the family Thermoproteaceae. The three strains were included in a single species due to high levels of DNA-DNA hybridization values. Based upon these results, the new isolates were assigned to a new genus and species in the family Thermoproteaceae. Thermocladium modestius gen. nov., sp. nov. The type strain is Thermocladium modestius IC-125T (= JCM 10088T). PMID- 9734042 TI - Identification of denitrifier strain T1 as Thauera aromatica and proposal for emendation of the genus Thauera definition. AB - Bacterial strain, T1, originally isolated by P.J. Evans on the basis of its capacity for toluene degradation under denitrifying conditions, has been classified as Thauera aromatica. In a comprehensive study of strains of this species, it was found that the cells have a different type of flagellar insertion from that of cells of the type species of the genus, Thauera selenatis, suggesting the convenience of an emendation of the description of the genus Thauera. Further studies on a larger collection of strains with the above characteristics may serve in the future as the basis for the creation of a new generic designation. PMID- 9734043 TI - Application of multiplex PCR using species-specific primers within the 16S rRNA gene for rapid identification of Nocardioides strains. AB - For the rapid identification of Nocardioides strains, multiplex PCR, using 16S rDNA as target gene, was used and its value was evaluated. Forward primers specific for Nocardioides albus, Nocardioides jensenii, Nocardioides plantarum and Nocardiodes simplex, among the five validly described Nocardioides species, were designed from the alignment of 165S rDNA sequences. Nocardioides luteus has been shown to be a member of the same species as N. albus by recent molecular systematic studies and preliminary DNA-DNA relatedness tests. Therefore, N. albus and N. luteus were considered as members of the same species in this study. Each primer was found to be species-specific by specificity testing. N. albus NSP01T, N. jensenii NSP19T, N. plantarum NSP21T and N. simplex NSP22T could be clearly differentiated by PCR products characteristics for each species in the multiplex PCR assays. N. luteus gave an identical results to N. albus NSP01. The additional 17 strains of N. albus and the additional four stains of N. simplex gave PCR products identical to those of N. albus NSP01T and N. simplex NSP22T, respectively. Multiplex PCR was found to be rapid, species-specific and reproducible. The technique evaluated in this study proved to be effective for rapidly identifying Nocardioides strains to species level. PMID- 9734044 TI - Nocardia flavorosea sp. nov. AB - An actinomycete strain, 'Nocardai flavorosea' JCM 3332, was found to have properties consistent with its classification in the genus Nocardia. An almost complete gene sequence of the 16S rDNA of the strain was determined following cloning and sequencing of the amplified gene. The sequence was aligned with those available for nocardiae and phylogenetic trees were inferred using four tree making algorithms. The organisms consistently formed a distinct clade with the type strain of Nocardia carnea. However, DNA relatedness experiments showed that the strain and N. carnea DSM 43397T belonged to two distinct genomic species. The organism was also distinguished from representative of all of the validly described species of Nocardia using a combination of phenotypic properties. These genotypic and phenotypic data show that the strain merits recognition as a new species of the genus Nocardia. The name proposed for the new species of is Nocardia flavorosea sp. nov. The type strain is JCM 3332T. PMID- 9734045 TI - Gordonia rhizosphera sp. nov. isolated from the mangrove rhizosphere. AB - The taxonomic position of bacterial strain 141T, isolated from the mangrove rhizosphere, has been clarified by phenotypic, chemotaxonomic and phylogenetic studies. The strain possesses wall chemotype IV, MK-9(H2) as the predominant menaquinone, relatively long-chain mycolic acids (56-64) carbon atoms) and straight-chain saturated and monounsaturated fatty acids with a small amount of tuberculostearic acid. The G+C content of the DNA is 66.8 mol%. Similarity values for genes encoding 16S rRNA indicated that strain 141T represents a new species within the genus Gordonia for which the name Gordonia rhizosphera sp. nov. is proposed. The type strain of G. rhizosphere is 141T (IFO 16068T). PMID- 9734046 TI - Methanococcus infernus sp. nov., a novel hyperthermophilic lithotrophic methanogen isolated from a deep-sea hydrothermal vent. AB - An autotrophic, extremely thermophilic methanogen (ME(T)) was isolated from a deep-sea hydrothermal chimney sample collected on the Mid-Atlantic Ridge at a depth of 3000 m. The heavily flagellated cells are motile and coccoid shaped. The new strain growths between 55 and 91 degrees C, with an optimum growth temperature at 85 degree C. The optimum pH for growth is 6.5, and the optimum sea salt concentration for growth is around 25 g l-1. The organism uses H2 and CO2 as the only substrate for growth and methane production. Tungsten, selenium and yeast extract stimulate growth significantly. In the presence of CO2 and H2, the organism reduces elemental sulphur to hydrogen sulphide. The G+C content of the genomic DNA is 33 mol%. As determined by 16S gene sequence analysis, this organism is closely related to Methanococcus jannaschii strain JAL-1T. However, no significant homology was observed between them with DNA-DNA hybridization. It is proposed that this organism should be placed in a new species, Methanococcus infernus. The type strain is ME(T) (= DSM 11812T). PMID- 9734047 TI - Streptococcus peroris sp. nov. and Streptococcus infantis sp. nov., new members of the Streptococcus mitis group, isolated from human clinical specimens. AB - Taxonomic studies were performed on eight strains of alpha-haemolytic streptococci that showed very low DNA-DNA hybridization similarity values with all established members of the mitis group of the genus Streptococcus. These strains were isolated from the tooth surface and pharynx of humans. 16S rRNA gene sequence analysis showed that these strains belonged to the mitis group, but that they fell into two new branches. DNA-DNA hybridization demonstrated two new similarity groups. From the results of the present study, the names Streptococcus peroris sp. nov. and Streptococcus infantis sp. nov. are proposed for these new groups. The type strains are O-66T (= GTC 848T = JCM 10158T) and O-122T (= GTC 849T = JCM 10157T), respectively. PMID- 9734048 TI - Actinobacillus scotiae sp. nov., a new member of the family Pasteurellaceae Pohl (1979) 1981 isolated from porpoises (Phocoena phocoena). AB - Phenotypic and phylogenetic studies were performed on a Gram-negative, rod-shaped bacterium isolated from three porpoises. Biochemical and physiological studies indicated that the bacterium was related to the family Pasteurellaceae. Comparative 16S rRNA gene sequencing studies confirmed these findings and demonstrated that the bacterium represents a hitherto unknown subline. The nearest phylogenetic relative of the unknown bacterium was Actinobacillus delphinicola, an organism also originating from sea mammals, although a sequence divergence of 3% demonstrated that the newly isolated bacterium is a distinct species. On the basis of the results of the phylogenetic analysis and phenotypic criteria, it is proposed that the bacterium should be classified as a new species, Actinobacillus scotiae sp. nov. The type strain of Actinobacillus scotiae sp. nov. is NCTC 12922T (= M2000/95/1T). PMID- 9734049 TI - Description of Acetobacter oboediens sp. nov. and Acetobacter pomorum sp. nov., two new species isolated from industrial vinegar fermentations. AB - Two strains of Acetobacter sp., LTH 2460T and LTH 2458T, have been isolated from running red wine and cider vinegar fermentations, respectively. Taxonomic characteristics of the isolates were investigated. Comparative analysis of the 165 rRNA sequences revealed > 99% similarity between strain LTH 2460T and the type strains of the related species Acetobacter europaeus and Acetobacter xylinus and between strain LTH 2458T and Acetobacter pasteurianus. On the other hand, low levels of DNA relatedness (< 34%) were determined in DNA-DNA similarity studies. This relatedness below the species level was consistent with specific physiological characteristics permitting clear identification of these strains within established species of acetic acid bacteria. Based on these results, the names Acetobacter oboediens sp. nov. and Acetobacter pomorum sp. nov. are proposed for strains LTH 2460T and LTH 2458T, respectively. The phylogenetic positions of the new species are reflected by a 16S rRNA-based tree. Furthermore, a 16S rRNA-targeted oligonucleotide probe specific for A. oboediens was constructed. PMID- 9734050 TI - Phylogenetic positions of phytoplasmas associated with dieback, yellow crinkle and mosaic diseases of papaya, and their proposed inclusion in 'Candidatus Phytoplasma australiense' and a new taxon, 'Candidatus Phytoplasma australasia'. AB - DNA extracted from three papaya (Carica papaya L.) plants, individually affected by dieback, yellow crinkle or mosaic diseases, was subjected to PCR using phytoplasma-specific primers to amplify the 16S rRNA gene plus 16S-23S rRNA intergenic spacer region. Near-complete DNA sequences obtained for the three PCR amplimers were subjected to phylogenetic analyses and direct sequence comparison with other phytoplasma 16S rDNA and 16S-23S spacer region DNA sequences. The papaya yellow crinkle (PpYC) and papaya mosaic (PpM) sequences were identical to each other, but distinctly different from the papaya dieback (PpDB) sequence, showing 90.3% identity in the he 16S rDNA and 87.8% identity in the 16S-23S spacer region DNA sequences. A phylogenetic tree based on 16S rDNA sequences was calculated, in which PpYC and PpM are most closely related to the tomato big bud phytoplasma (TBB; 99.7% 16S rDNA sequence identity) from Australia, within subclade iii. This subclade consists of strains only reported occurring in the Southern Asian region and Australia, which indicates an Asian/Australasian origin. PpDB is most closely related to the Phormium yellow leaf phytoplasma from new Zealand (PYL; 99.9% identity) and the Australian grapevine yellows phytoplasma (AGY; 99.7% identity). These three phytoplasma strains form a distinct clade within subclade xii, which also includes the European strains STOL and VK as another distinct clade. The origin of the closely related but geographically separated AGY-like strains and STOL-like strains of subclade xii is unclear. It is proposed that phytoplasma strains PpDB, PYL and AGY be included in the previously described taxon 'Candidatus Phytoplasma australiense', and that PbYC, PpM and TBB be assigned to a new taxon, "Candidatus Phytoplasma australasia'. PMID- 9734051 TI - Azoarcus anaerobius sp. nov., a resorcinol-degrading, strictly anaerobic, denitrifying bacterium. AB - A strictly anaerobic, nitrate-reducing bacterium, strain LuFRes1, was isolated using resorcinol as sole source of carbon and energy. The strain reduced nitrate to dinitrogen gas and was not able to use oxygen as an alternative electron acceptor. Cells were catalase-negative but superoxide-dismutase-positive. Resorcinol was completely oxidized to CO2. 16S rRNA sequence analysis revealed a high similarity with sequences of Azoarcus evansii and Azoarcus tolulyticus. Strain LuFRes1T (= DSM 12081T) is described as a new species of the genus Azoarcus, Azoarcus anaerobius. PMID- 9734052 TI - Taxonomic rearrangements of the genera Thiocapsa and Amoebobacter on the basis of 16S rDNA sequence analyses, and description of Thiolamprovum gen. nov. AB - Complete nucleotide sequences of the 16S rDNAs were determined from Thiocapsa and Amoebobacter species, including all available type strains and some additional isolates. The distance-matrix analysis and the dendrogram for estimating the genetic relationships revealed that the investigated strains were found in two major clusters within the Chromatiaceae. One cluster comprises all Amoebobacter species, Thiocapsa roseopersicina and several isolates related to Thiocapsa roseopersicina. Representatives of the species Amoebobacter roseus, Amoebobacter pendens and Thiocapsa roseopersicina, the so called 'Thiocapsa roseopersicina group', are very closely related, justifying their inclusion into one genus, Thiocapsa, for which an emended description is presented. Amoebobacter purpureus and Amoebobacter pedioformis formed two separate lines of descent with less than 93% (89.6-92.9%) similarity to strains of the 'Thiocapsa roseopersicina group'. Therefore, they will be considered as two separate genera. As a consequence, an emended description is presented for the genus Amoebobacter, with Amoebobacter purpureus as the new type species and A. pedioformis is transferred to Thiolamprovum pedioforme gen. nov., comb. nov. Two species, Thiocapsa pfennigii and Thiocapsa halophila, which have been classified with the genus Thiocapsa because of their morphological properties, were found within another major cluster of the Chromatiaceae and are only distantly phylogenetically related to the first cluster with 88.4-90.6% and 90.4-92.2% sequence similarity, respectively. PMID- 9734053 TI - Shewanella amazonensis sp. nov., a novel metal-reducing facultative anaerobe from Amazonian shelf muds. AB - A new bacterial species belonging to the genus Shewanella is described on the basis of phenotypic characterization and sequence analysis of its 16S rRNA encoding and gyrase B (gyrB) genes. This organism, isolated from shallow-water marine sediments derived from the Amazon River delta, is a Gram-negative, motile, polarly flagellated, facultatively anaerobic, rod-shaped eubacterium and has a G&C content of 51.7 mol%. Strain SB2BT is exceptionally active in the anaerobic reduction of iron, manganese and sulfur compounds. SB2BT grows optimally at 35 degrees C, with 1-3% NaCl and over a pH range of 7-8. Analysis of the 16S rDNA sequence revealed a clear affiliation between strain SB2BT and members of the gamma subclass of the class Proteobacteria. High similarity values were found with certain members of the genus Shewanella, especially with Shewanella putrefaciens, and this was supported by cellular fatty acid profiles and phenotypic characterization. DNA-DNA hybridization between strain SB2BT and its phylogenetically closest relatives revealed low similarity values (24.6-42.7%) which indicated species status for strain SB2BT. That SB2BT represents a distinct bacterial species within the genus Shewanella is also supported by gyrB sequence analysis. Considering the source of the isolate, the name Shewanella amazonensis sp. nov. is proposed and strain SB2BT (= ATCC 700329T) is designated as the type strain. PMID- 9734054 TI - Proposal of six new species in the genus Microbacterium and transfer of Flavobacterium marinotypicum ZoBell and Upham to the genus Microbacterium as Microbacterium maritypicum comb. nov. AB - Reference strains, including two mis-named organisms, 'Chromobacterium chocolatum' and Flavobacterium marinotypicum, isolates from soil and clinical specimens, all previously recognized as Aureobacterium or Microbacterium, were characterized taxonomically. On the basis of morphological, physiological and chemotaxonomic characteristics, as well as DNA-DNA hybridization data, six new species and one new combination are proposed in the genus Microbacterium: Microbacterium ketosireducens sp. nov. (type strain IFO 14548T), Microbacterium chocolatum sp. nov. (type strain IFO 3758T), Microbacterium aurantiacum sp. nov. (type strain IFO 15234T), Microbacterium hominis sp. nov. (type strain IFO 15708T), Microbacterium thalassium sp. nov. (type strain IFO 16060T), Microbacterium halophilum sp. nov. (type strain IFO 16062T) and Microbacterium maritypicum comb. nov. (type strain IFO 15779T). PMID- 9734055 TI - Tissierella creatinophila sp. nov., a gram-positive, anaerobic, non-spore forming, creatinine-fermenting organism. AB - A strictly anaerobic, Gram-positive, non-spore-forming bacterium was isolated from sewage sludge which grew on creatinine as sole source of carbon and energy. This new isolate, designated strain KRE 4T, totally degraded creatinine via creatine, sarcosine and glycine to the products acetate, monomethylamine, ammonia and carbon dioxide. Growth on creatinine or creatine was selenium-dependent and stimulated by formate, indicating the involvement of a creatine reductase, sarcosine reductase and/or glycine reductase. This was substantiated by the fact that creatine, sarcosine and glycine were reduced by cell-free extracts. Growth on creatinine or creatine was also possible in the absence of formate, but with an increase in doubling time. The new bacterium occurred as rod-shaped cells, which exhibited an angular form (2-6 microns long and 0.7-1.1 microns wide) and showed motility by means of peritrichous flagella. The G+C content of the DNA was 30 mol %. Comparative 16S rRNA sequence analysis demonstrated that strain KRE 4T represents a new subline within the genus Tissierella. Due to its very restricted substrate spectrum and the inability of whole cells to utilize sarcosine and glycine as intermediates of creatine breakdown, this organism can be readily separated from currently described species of Tissierella. Therefore, based on the phenotypic and phylogenetic distinctiveness of the new isolate, it si proposed that the bacterium be classified as a new species of the genus Tissierella, Tissierella creatinophila sp. nov. The type strain is KRE 4 (= DSM 6911T). PMID- 9734056 TI - A new genus of the order Actinomycetales, Cryptosporangium gen. nov., with descriptions of Cryptosporangium arvum sp. nov. and Cryptosporangium japonicum sp. nov. AB - Four strains that form sporangia with motile sporangiospores and mycelia were isolated from soil samples. Their many sporangia were covered by mycelia. They had glutamic acid, glycine, alanine and meso-diaminopimelic acid as cell wall amino acids (wall chemotype II), acofriose (3-O-mythylrhamnose) as a characteristic whole-cell sugar, and menaquinone 9(H6). The taxonomic characteristics of these strains differ from those of the previously described motile actinomycetes. On the basis of the morphological, physiological, chemotaxonomic and phylogenetic analyses, a new genus is proposed, Cryptosporangium, and two new species, Cryptosporangium arvum sp. nov (type strain IFO 15965T) for strain YU 629-21T and Cryptosporangium japonicum sp. nov (type strain IFO 15966T) for strains YU 636-3T, YU 655-31 and YU 656-31. PMID- 9734057 TI - Petrotoga mobilis sp. nov., from a North Sea oil-production well. AB - Rod-shaped, thermophilic bacteria with a sheath-like outer structure (toga) were isolated from hot oilfield water of a North Sea oil reservoir. One of the isolates, designated SJ95(T), is an obligately anaerobic, sheathed, Gram negative, fermentative bacterium capable of reducing elemental sulfur to hydrogen sulfide and tolerating high salt concentrations. The optimum growth conditions for this isolate are 58-60 degrees Celsius and pH 6.5-7.0 with 3-4% NaCl and 0.7% MgSO(4). 7H(2)O in the medium. Vitamins are required for growth. Growth is stimulated by yeast extract. Cells of strain SJ95(T) vary in size from 1-2 to 40 50 micron in length and are motile with a subpolar flagellation. Cels grown on xylan have xylanase activity, presumably associated with the toga, and glucose isomerase activity was detected in xylose-grown cells. The DNA G+C content is 31 and 34 mol%, determined by the thermal denaturation and HPLC methods, respectively. Phylogenetically, strain SJ95(T) is most closely related to Petrotoga miotherma with a 97.7% similarity level between their 165 rDNA sequences. The DNA-DNA reassociation value between the two DNAs was 35.6%. On the basis of differences in genotypic, phenotypic and immunological characteristics, strain SJ95t (=DSM 10674t) is proposed as the type strain of a new species, Petrotoga mobilis. It can be readily distinguished from P. miotherma by its motility. PMID- 9734058 TI - Structure and genetic stability of mitochondrial genomes vary among yeasts of the genus Saccharomyces. AB - Several yeast species/isolates belonging to the genus Saccharomyces were examined for the organization of their mtDNAs and ability to generate petite mutants. A general characteristic for all of the mtDNAs tested was that they were very A+T rich. However, restriction patterns and inducibility of petite mutations revealed a great diversity in the organization and genetic behaviour of mtDNAs. One group of yeasts, Saccharomyces sensu stricto, contains mtDNA ranging in size from 64 to 85 kb. mtDNAs form these yeasts contain a high number of restriction sites that are recognized by the enzymes Haelll and Mspl, which cut specifically in G+C clusters. There are three to nine ori/rep sequences per genome. These yeasts spontaneously generate respiration deficient mutants. Ethidium bromide (Et-Br), at low concentrations, induces a majority of cells to give rise to petites. A second group of yeasts, Saccharomyces sensu lato, contains smaller mtDNAs, ranging in size from 23 to 48 kb, and probably only a few intergenic G+C clusters and no ori/rep sequences. These yeasts also generate petite clones spontaneously. but Et-Br, even when present at high concentrations, does not substantially increase the frequency of petites. In most petite clones from these yeasts only a small fragment of the wild-type molecule is retained and apparently multiplied. A third group, represented by Saccharomyces kluyveri, does not give rise to petite mutants either spontaneously or after induction. PMID- 9734059 TI - Bullera penniseticola sp. nov. and Kockovaella sacchari sp. nov., two new yeast species isolated from plants in Thailand. AB - Two strains of ballistocondium-forming yeasts, isolated from plants collected in the south-east seacoast of Bangkok, Thailand, were described. The strains (K 272(T) and K-337(T)) were assigned to the genera Bullera and Kockovaella, respectively, based on morphological and chemotaxonomical characteristics. Phylogenetically, strain K-272(T) is close to Bullera hannae, and strain K-337(T) is close to Kockovaella thailandica and Kockovaella imperatae. These two strains represent new species based on DNA-DNA reassociation experiments. Bullera penniseticola Takashima et Nakase sp. nov. and Kockovaella sacchari Takashima et Nakase sp. nov. are proposed for K-272(T) (=JCM 9857(T)) and K-337(T) (=JCM 9858(T)), respectively. PMID- 9734060 TI - Bacillus pseudomycoides sp. nov. AB - Previous DNA relatedness studies showed that strains identified as Bacillus mycoides segregated into two genetically distinct yet phenotypically similar groups, one being B. mycoides sensu stricto and the other, an unclassified taxon. In the present study, the taxonomic position of this second group was assessed by measuring DNA relatedness and determining phenotypic characteristics of an increased number of B. mycoides strains. Also determined was the second group's 16S RNA gene sequence. The 36 B. mycoides strains studied segregated into two genetically distinct groups showing DNA relatedness of about 30%; 18 strains represented the species proper and 18 the second group with intragroup DNA relatedness for both groups ranging from 70 to 100%. DNA relatedness to the type strains of presently recognized species with G+C contents of approximately 35 mol% (Bacillus alcalophilus, Bacillus cereus, Bacillus circulans, Bacillus lentus, Bacillus megaterium and Bacillus sphaericus) ranged from 22 to 37%. Although shown to be genetically distinct taxa, the two B. mycoides groups exhibited highly similar (98%) 16S RNA sequences. Phylogenetic analyses showed that both B. mycoides and the second group clustered closely with B. cereus. Although not distinguishable by physiological and morphological characteristics, the two B. mycoides groups and B. cereus were clearly separable based on fatty acid composition. The data established that the second B. mycoides group merits recognition as a new species for which the name Bacillus pseudomycoides is proposed. The type stain is NRRL B-617(T). PMID- 9734061 TI - Pseudoalteromonas prydzensis sp. nov., a psychrotrophic, halotolerant bacterium form Antarctic sea ice. AB - Species of the genus Pseudoalteromonas are frequently isolated from marine ecosystems and appear to be particularly abundant in Antarctic coastal waters. Most Pseudoalteromonas strains isolated form sea ice and underlying seawater samples are phenotypically similar to the species Pseudoalteromonas antarctica and Pseudoalteromonas nigrifaciens. However, a minority of isolates were recognized by phenotypic, DNA-DNA hybridization and 16S rRNA-based phylogenetic studies to represent a distinct genospecies clustering at the periphery of the non-pigmented, Pseudoalteromonas species clade. These strains are non-pigmented, halotolerant psychrotrophs that are capable of hydrolysing starch and chitin, and possess a DNA G+C content of 38-39 mol%. It is proposed that this group represents a novel species, Pseudoalteromonas prydzensis sp. nov., for which the type strain is ACAM 620(T). PMID- 9734062 TI - Proposal of Craurococcus roseus gen. nov., sp. nov. and Paracraurococcus ruber gen. nov., sp. nov., novel aerobic bacteriochlorophyll a-containing bacteria from soil. AB - Sequences of the 16S rRNA gene were determined for three strains of aerobic bacteriochlorophyll a-containing bacteria isolated from soil. The sequence of two strains (NS89(T) and NS102) were identical for approximately 1500 nucleotides. Phylogenetic analysis revealed that the three strains belonged to the alpha-1 subclass of the Proteobacteria, constituting one line of descent. The three strains are comparatively related to Roseoccus thiosulfatophilus, which is an aerobic bacteriochlorophyll a-containing bacterium. The 16S rRNA gene sequence similarity and the DNA-DNA relatedness allow the proposal of two new genera, Craurococcus gen. nov. and Paracraurococcus gen. nov. The type species are Craurococus roseus sp. nov. and Paracraurococcus ruber sp. nov., and their type strains are NS130(T) (=JCM 9933(T)) and NS89(T) (=JCM 9931(T)), respectively. PMID- 9734063 TI - Identification of Staphylococcus species by 16S-23S rDNA intergenic spacer PCR analysis. AB - To investigate whether 16S-23S rDNA (rDNA) spacer region length polymorphisms are suitable for the identification of Staphylococcus strains, the 16S-23S rDNA intergenic spacer region lengths of 221 strains belonging to 31 species were studied by using a PCR-based method. Each species presented a specific 16S-23S pattern made of 1-8 fragments ranging from 104-771 bp, with the exception of the species Staphylococcus warnei, Staphylococcus caprae and Staphylococcus piscifermentans, which presented larger or smaller fragments. Very few species showed more than one pattern, Staphylococcus saprophyticus subsp. saprophyticus and Staphylococcus aureus being the most heterogeneous species (five different patterns for eight strains). Five clinical strains that could not be identified at the species level by phenotypical tests were finally identified using this method. Discrimination between some species that showed close patterns (Staphylococcus aureus/Staphylococcus chromogenes/Staphylococcus equorum, Staphylococcus aureus/staphylococcus intermedius, Staphylococcus delphini/Staphylococcus felis, Staphylococcus gallinarum, Staphylococcus delphini/Staphylococcus felis, Staphylococcus vitulus/Staphylococcus auricularis) was further achieved after Dral digestion of the PCR products. Although it does not allow discrimination of subspecies, the use of 16S-23S spacer region length data determined by PCR-mediated amplification is suitable for the identification of the 31 Staphylococcus species tested in this study. The method is rapid, easy and may be a useful tool for the identification of Staphylococcus species in the clinical microbiology laboratory. PMID- 9734064 TI - Characterization and reclassification of an aromatic- and chloroaromatic degrading Pseudomonas sp., strain HV3, as Sphingomonas sp. HV3. AB - Phylogenetic analyses of 16S rRNA gene sequences showed that the Gram-negative aromatic- and chloroaromatic-degrading Pseudomonas sp. strain HV3 carrying the mega-plasmid pSKY4 belongs to the genus Sphingomonas. The 16SrRNA sequence is most related to Sphingomonas chlorophenolica strains ATCC 33790(T) (98.5%) and SR3 (98.4%) and Sphingomonas sp. SS86 (98.4%). The G+C content was 64 mol%, and the DNA-DNA hybridization-based relative homology of strain HV3 to the S. chlorophenolica ATCC 33790(T) and S. chlorophenolica RA2 was 59.6% and 35.9%, respectively. The results showed that although strain HV3 is related to S. chlorophenolica it differs in certain characteristics. It is therefore proposed to reclassify Pseudomonas sp. strain HV3 as Sphingomonas sp. HV3. PMID- 9734065 TI - 16S rDNA sequence variations of some Streptococcus suis serotypes. AB - Streptococcus suis 16S rDNA from selected serotypes has been sequenced and compared with the 16S rDNA sequences from serotypes 1 and 2 present in Genbank. After alignment the sequenced serotypes show clusters of variation. Based on these clusters, a limited phylogenetic tree showing the relationships of all of the serotypes was constructed. PMID- 9734066 TI - DNA sequencing reveals limited heterogeneity in the 16S rRNA gene from the rrnB operon among five Mycoplasma hominis isolates. AB - To investigate the intraspecies heterogeneity within the 16S rRNA gene of Mycoplasma hominis, five isolates with diverse antigenic profiles, variable/identical P120 hypervariable domains, and different 16S rRNA gene RFLP patterns were analysed. The 16S rRNA gene from the rrnB operon was amplified by PCR and the PCR products were sequenced. Three isolates had identical 16S rRNA sequences and two isolates had sequences that differed from the others by only one nucleotide. PMID- 9734067 TI - Morphology of the brain and sense organs in the snailfish Paraliparis devriesi: neural convergence and sensory compensation on the Antarctic shelf. AB - The Antarctic snailfish Paraliparis devriesi (Liparidae) is an epibenthic species, inhabiting depths of 500-650 m in McMurdo Sound. Liparids are the most speciose fish family in the Antarctic Region. We examine the gross morphology and histology of the sense organs and brain of P. devriesi and provide a phyletic perspective by comparing this morphology to that of four scorpaeniforms and of sympatric perciform notothenioids. The brain has numerous derived features, including well-developed olfactory lamellae with thick epithelia, large olfactory nerves and bulbs, and large telencephalic lobes. The retina contains only rods and exhibits a high convergence ratio (82:1). Optic nerves are small and nonpleated. The tectum is small. The corpus of the cerebellum is large, whereas the valvula is vestigial. The rhombencephalon and bulbospinal junction are extended and feature expanded vagal and spinal sensory lobes as well as hypertrophied dorsal horns and funiculi in the rostral spinal cord. The lower lobes of the pectoral fins have taste buds and expanded somatosensory innervation. Although the cephalic lateral line and anterior lateral line nerve are well developed, the trunk lateral line and posterior lateral line nerve are reduced. Near-field mechanoreception by trunk neuromasts may have been compromised by the watery, gelatinous subdermal extracellular matrix employed as a buoyancy mechanism. The expanded somatosensory input to the pectoral fin may compensate for the reduction in the trunk lateral line. The brains of P. devriesi and sympatric notothenioids share well-developed olfactory systems, an enlarged preoptic-hypophyseal axis, and subependymal expansions. Although the functional significance is unknown, the latter two features are correlated with habitation of the deep subzero waters of the Antarctic shelf. PMID- 9734068 TI - Histological studies of the dorsal nasal, angularis oculi, and facial veins of sheep (Ovis aries). AB - Selective brain cooling (SBC) requires vasoactivity in the superficial veins of the face of the animal. This vasoactivity is possible because of an adequate amount of smooth muscle in the tunica media of each of these superficial vessels, enabling it to act as a "muscle sphincter". In this study, the angularis oculi, dorsal nasal, distal, and proximal parts of the facial veins in sheep were examined histologically to describe an anatomical basis for SBC. Measurements of the tunica media thickness, the lumen diameter, and the ratio of these measurements showed that the relative tunica media thicknesses in the angularis oculi vein and the dorsal nasal vein are statistically smaller (P < 0.001) than in the distal or the proximal parts of the facial vein. In the angularis oculi, dorsal nasal, and distal part of the facial vein, the tunicae mediae were composed of five to seven circularly arranged smooth muscle layers, suggesting their ability to vasoconstrict. The proximal part of the facial vein possesses both circularly and longitudinally arranged smooth muscle layers. The circular smooth muscle layers suggest a vasoconstrictory function, whereas the longitudinal smooth muscle layers suggest a vasoconstrictory function in this part of the facial vein. Both the dorsal nasal and the proximal part of the facial vein, but not the angularis oculi or the distal part of the facial vein, possess endothelial valves near their confluences with other veins. It was concluded from this study that the angularis oculi and the distal part of the facial vein vasoconstrict, whereas the proximal part of the facial vein vasodilates, enabling the necessary changes in blood flow in SBC. PMID- 9734069 TI - Sequence analysis of the mitochondrial DNA control region of ciscoes (genus Coregonus): taxonomic implications for the Great Lakes species flock. AB - Sequence variation in the control region (D-loop) of the mitochondrial DNA (mtDNA) was examined to assess the genetic distinctiveness of the shortjaw cisco (Coregonus zenithicus). Individuals from within the Great Lakes Basin as well as inland lakes outside the basin were sampled. DNA fragments containing the entire D-loop were amplified by PCR from specimens of C. zenithicus and the related species C. artedi, C. hoyi, C. kiyi, and C. clupeaformis. DNA sequence analysis revealed high similarity within and among species and shared polymorphism for length variants. Based on this analysis, the shortjaw cisco is not genetically distinct from other cisco species. PMID- 9734071 TI - 'Floral' scent production by Puccinia rust fungi that mimic flowers. AB - Crucifers (Brassicaceae) in 11 genera are often infected by rust fungi in the Puccinia monoica complex. Infection causes a 'pseudoflower' to form that is important for attracting insect visitors that sexually outcross the fungus. 'Pollinator' attraction is accomplished through visual floral mimicry, the presence of a nectar reward and floral fragrances. Here we used gas chromatography and mass spectrometry to identify and quantify fragrance production by these rust fungi on several Arabis hosts, and by co-occurring true flowers that share insect visitors. Fungal pseudoflowers produced distinctive floral fragrances composed primarily of aromatic alcohols, aldehydes and esters. Pseudoflower fragrances were chemically similar to noctuid-moth-pollinated flowers, such as Cestrum nocturnum and Abelia grandiflora, but were very different from host flowers, host vegetation and the flowers of coblooming, nonhost angiosperms. There was variation in the quantity and composition of fragrance profiles from different fungal species as well as within and among hosts. The evolution of scent chemistry is relatively conservative in these fungi and can be most parsimoniously explained in three steps by combining chemical data with a previously determined rDNA ITS sequence-based phylogeny. Pseudoflower scent does not appear to represent a simple modification of host floral or vegetative emissions, nor does it mimic the scent of coblooming flowers. Instead, we suspect that the unique fragrances, beyond their function as pollinator attractants, may be important in reducing gamete loss by reinforcing constancy among foraging insects. PMID- 9734070 TI - Assessing hybridization in natural populations of Penstemon (Scrophulariaceae) using hypervariable intersimple sequence repeat (ISSR) bands. AB - Inferences regarding hybridization rely on genetic markers to differentiate parental taxa from one another. Intersimple sequence repeat (ISSR) markers are based on single-primer PCR reactions where the primer sequence is derived from di and trinucleotide repeats. These markers have successfully been used to assay genetic variability among cultivated plants, but have not yet been tested in natural populations. We used genetic markers generated from eight ISSR primers to examine patterns of hybridization and purported examples of hybrid speciation in Penstemon (Scrophulariaceae) in a hybrid complex involving P. centranthifolius, P. grinnellii, P. spectabilis and P. clevelandii. This hybrid complex has previously been studied using three molecular data sets (allozymes, and restriction-site variation of nuclear rDNA and chloroplast DNA). These studies revealed patterns of introgression involving P. centranthifolius, but were unsuccessful in determining whether gene flow occurs among the other species, and support for hypotheses of diploid hybrid speciation was also lacking. In this study, we were able to fingerprint each DNA accession sampled with one to three ISSR primers and most accessions could be identified with a single primer. We found population- and species-specific markers for each taxon surveyed. Our results: (i) do not support the hybrid origin of P. spectabilis; (ii) do support the hypothesis that P. clevelandii is a diploid hybrid species derived from P. centranthifolius and P. spectabilis; and (iii) demonstrate that pollen-mediated gene flow via hummingbird vectors is prevalent in the hybrid complex. PMID- 9734072 TI - Phylogeography of the West Indian manatee (Trichechus manatus): how many populations and how many taxa? AB - To resolve the population genetic structure and phylogeography of the West Indian manatee (Trichechus manatus), mitochondrial (mt) DNA control region sequences were compared among eight locations across the western Atlantic region. Fifteen haplotypes were identified among 86 individuals from Florida, Puerto Rico, the Dominican Republic, Mexico, Columbia, Venezuela, Guyana and Brazil. Despite the manatee's ability to move thousands of kilometers along continental margins, strong population separations between most locations were demonstrated with significant haplotype frequency shifts. These findings are consistent with tagging studies which indicate that stretches of open water and unsuitable coastal habitats constitute substantial barriers to gene flow and colonization. Low levels of genetic diversity within Florida and Brazilian samples might be explained by recent colonization into high latitudes or bottleneck effects. Three distinctive mtDNA lineages were observed in an intraspecific phylogeny of T. manatus, corresponding approximately to: (i) Florida and the West Indies; (ii) the Gulf of Mexico to the Caribbean rivers of South America; and (iii) the northeast Atlantic coast of South America. These lineages, which are not concordant with previous subspecies designations, are separated by sequence divergence estimates of d = 0.04-0.07, approximately the same level of divergence observed between T. manatus and the Amazonian manatee (T. inunguis, n = 16). Three individuals from Guyana, identified as T. manatus, had mtDNA haplotypes which are affiliated with the endemic Amazon form T. inunguis. The three primary T. manatus lineages and the T. inunguis lineage may represent relatively deep phylogeographic partitions which have been bridged recently due to changes in habitat availability (after the Wisconsin glacial period, 10 000 B P), natural colonization, and human-mediated transplantation. PMID- 9734073 TI - Mitochondrial DNA phylogeography of European hedgehogs. AB - European hedgehog populations belonging to Erinaceus europaeus and E. concolor have been investigated by mitochondrial DNA analysis. A 383 bp fragment of the cytochrome b gene has been sequenced and maximum parsimony and neighbour-joining trees of Tamura-Nei genetic distance values have been constructed. Similar topologies have been produced by both methods, showing a deep divergence between E. europaeus and E. concolor and a further subdivision of each species into a western and an eastern clade. A comparison with previously published allozyme data is made, and concordant and discordant patterns are discussed. The influence of Pleistocene glaciations on the observed pattern of divergence is inferred. PMID- 9734074 TI - Genetic variation and bill size dimorphism in a passerine bird, the reed bunting Emberiza schoeniclus. AB - In passerine birds morphological differentiation in bill size within species is not commonly observed. Bill size is usually associated with a trophic niche, and strong differences in it may reflect the process of genetic differentiation and, possibly, speciation. We used both mitochondrial DNA (mtDNA) and nuclear microsatellites to study genetic variation between two subspecies of reed bunting, Emberiza schoeniclus schoeniclus and E.s. intermedia, along their distributional boundary in western Europe. These two subspecies are characterized by a high dimorphism in bill size and, although breeding populations of the two subspecies are found very close to each other in northern Italy, apparently no interbreeding occurs. The observed morphological pattern between the two subspecies may be maintained by geographically varying selective forces or, alternatively, may be the result of a long geographical separation followed by a secondary contact. MtDNA sequences of cytochrome b and ND5 (515 bp) showed little variation and did not discriminate between the two subspecies, indicating a divergence time of less than 500 000 years. The analysis of four microsatellite loci suggested a clear, although weak, degree of genetic differentiation in the large- and small-billed populations, as indicated by FST and RST values and genetic distances. The correlation between bill size and genetic distance between populations remained significant after accounting for the geographical distances between sampling localities. Altogether, these results indicate a very recent genetic differentiation between the two bill morphs and suggest that a strong selection for large bills in the southern part of the breeding range is probably involved in maintaining the geographical differentiation of this species. PMID- 9734075 TI - Phylogeography of skinks (Chalcides) in the Canary Islands inferred from mitochondrial DNA sequences. AB - Mitochondrial DNA (mtDNA) evolution was investigated in skinks of the genus Chalcides found in the Canary Islands (Ch. sexlineatus, Ch. viridanus and Ch. simonyi), together with some North African congenerics (Ch. polylepis and Ch. mionecton). Several sites were included within islands to cover areas of known within-island geographical variation in morphology. Skinks from the islands of El Hierro and La Gomera appear to be sister taxa. The relationships between this clade and the Tenerife and Gran Canarian skinks were not fully resolved, although the best working hypothesis indicated monophyly with the former, with the latter forming a closely related outgroup. Ch. simonyi from Fuerteventura was more distantly related to the Western Canary Island skinks and did not show close relationships with the North African species Ch. mionecton and Ch. polylepis. Possible colonization sequences for the four most Western Canary Islands were considered. El Hierro appears to have been colonized relatively recently from La Gomera, commensurate with the recent origin of this island, while dispersal between La Gomera and Tenerife and between Gran Canaria and Tenerife or La Gomera appears to have taken place considerably earlier. Substantial within-island haplotype divergence was found in Gran Canaria and Tenerife. This may be a result of recent periods of intense volcanic activity found within these two islands. Lower levels of within-island differentiation are found in La Gomera and El Hierro and may be explained by lower levels of volcanic activity during recent geological history and a more recent colonization, respectively. PMID- 9734076 TI - Evidence for transfer of antibiotic-resistance genes in soil populations of streptomycetes. AB - Phylogenetic analysis was used to evaluate the hypothesis of gene transfer in streptomycetes, many of which are antibiotic producers. The diversity and possible origins of streptomycin-resistance genes was investigated for a population of Streptomyces strains isolated from a site in Brazil where antibiotic production had previously been implicated The analysis provides compelling evidence for the transfer of these genes. Examination of other Streptomyces-type strains also reveals a scattered distribution of streptomycin producers with respect to the overall phylogeny. These results suggest that horizontal gene transfer may be an important factor in the evolution of antibiotic genes in streptomycetes. PMID- 9734078 TI - Tissue boiling: a short-cut in DNA extraction for large-scale population screenings. PMID- 9734077 TI - Density and variability of dinucleotide microsatellites in the parthenogenetic polyploid snail Melanoides tuberculata. AB - Characterization of microsatellites in the parthenogenetic polyploid snail Melanoides tuberculata revealed an unusual high density of dinucleotide repeats. Multiple banding patterns were obtained at these loci, and interpreted as a consequence of polyploidy. Microsatellite variability was low within, but high between, shell morphotypes. Genotypes were wholly transmitted from mothers to offspring. These results suggest that reproduction is strictly apomictic, and that shell morphotypes are genetic clones. PMID- 9734079 TI - Polymorphic trinucleotide microsatellite loci for a neotropical parrot, the green rumped parrotlet, Forpus passerinus. PMID- 9734080 TI - Microsatellite primers for the Eurasian otter. PMID- 9734081 TI - Charaterization of (GT)n microsatellites from native white shrimp (Penaeus setiferus). PMID- 9734082 TI - Isolation and characterization of microsatellite markers in the periwinkle Littorina striata King & Broderip, 1832 (Mollusca, Gastropoda, Prosobranchia). PMID- 9734083 TI - Universal PCR primers for S7 ribosomal protein gene introns in fish. PMID- 9734085 TI - Isolation and characterization of highly polymorphic microsatellites in the water vole, Arvicola terrestris. PMID- 9734084 TI - Genetic distinction of scorpionflies (Panorpa vulgaris) by microsatellites. PMID- 9734086 TI - Characterization of microsatellite loci in Pinus sylvestris L. PMID- 9734087 TI - Characterization of tetranucleotide microsatellite markers in the Scottish crossbill (Loxia scotica). PMID- 9734089 TI - In memory of Gottfried Mollenstedt. PMID- 9734088 TI - Electron transfer in the photosynthetic reaction center: mechanistic implications of mutagenesis studies. AB - A phenomenological analysis of the driving force effects in photosynthetic reaction centers modified by mutagenesis and also by chemical means is presented. Different parameter sets associated with different mechanisms of electron transfer are consistent with the mutagenesis experiments. However, only one parameter set--connected with a sequential mechanism of electron transfer--is consistent with all known experimental data. Arguments explaining why the sequential mechanism of electron transfer is selected by nature in the wild type reaction center are provided. Why the driving force of the wild type reaction center is about 0.25 eV is explained and new driving force effects are predicted. PMID- 9734090 TI - [Surgical treatment of thyroid diseases in children]. AB - In order to review the experience of a single institution with thyroid surgery and identify prognostic factors, we did a retrospective chart review of all patients who underwent thyroid surgery between 1980 and 1995. Fifty patients, 32 girls and 18 boys, underwent thyroid surgery. Clinical presentation include a thyroid nodule (36 patients), a family history of MEN syndrome with a positive pentagastrin stimulation test (10), diffuse nodular goitre (3) and a neonatal cervical mass (1). Radiologic investigation include ultrasonography in 58% of cases, radionucleide imaging in 70% of cases. The mean age at surgery was 13.3 years and the delay between onset of symptoms and diagnostic was 13 months. Twenty-one patients had a lobectomy +/- isthmectomy, 19 had a total thyroidectomy and 10 had a subtotal thyroidectomy. Nine patients required a second surgery to complete the surgical treatment, and perform a total or near total thyroidectomy when the final pathology showed a carcinoma. A well differentiated carcinoma was found in 52% of the patients, a medullary carcinoma in 20% and a benign lesion in 28% of cases. Nine patients had local or distant metastases at initial surgery. Post-op treatment consisted of suppressive hormonotherapy in all cases of cancer and radioactive iodine when recurrence occurred in 24% of patients and when metastasis where present. Survival remained at 100%. CONCLUSION: Thyroid nodules should be rapidly investigated in children because of a high potential of cancerous lesions. With an aggressive surgical treatment and post-op I131 for recurrent lesions and metastasis the prognostic remains excellent. PMID- 9734091 TI - [Intrahepatic glutathione and oxidative stress in liver transplantation in the pig]. AB - OBJECT: To determine the loss of endogenous GSH from livers cold-stored and reperfused, using a model of liver transplantation in the pig. MATERIAL AND METHODS: Four female Yorkshire pigs weighing 19 to 40 kg received a liver allograft. Donor livers were cold-stored in the UW solution. Mean cold ischemic time was 6.5 hours. Malondialdehyde (MDA) levels were used as an index of oxidative stress. MDA plasma levels were measured following recipient laparotomy (H0), immediately (H1), and 90 minutes after liver reperfusion (H2). MDA and GSH levels in liver were measured following donor laparotomy (T0), at the end of cold ischemic period (T1), and at 90 minutes following liver reperfusion (T2). RESULTS: Three animals survived. MDA liver levels decreased of 44% between T0 and T1, then increased to 92% at T2. In contrast, in plasma, graft reperfusion was associated with an increase of MDA to 140% of the baseline values which reached 188% at H2. Intrahepatic GSH levels decreased of 49% at T1, then to 72% at T2. CONCLUSION: our study suggests that in liver transplantation: (1) Hepatic GSH is depleted to 49% during cold-storage, and an additional 23% is lost after reperfusion; (2) GSH contained in the UW solution does not prevent the loss of hepatocellular glutathione during preservation and reperfusion; (3) after short periods of cold ichemia, endogenous hepatic GSH may protect against oxydative stress in the transplanted liver. PMID- 9734093 TI - [Intra-observer variability of measurement of posture with three-dimensional digitization]. AB - A new system based on 3D digitization with magnetic fields has been developed by our research team. The focus of this study was to evaluate the intratester reproducibility of this technique of measurement. Twenty key morphological parameters were used twice to digitize the fourty five female subjects aged 7 to 23 years. The results of variance analysis (ANOVA) for repeated measures showed no statistically significant difference between the two series of measurements for the twenty angles studied. In 56% of the measurements, the difference of the means was less than 1 degree, the greatest being 1.82 degrees. These results confirm the reliability of this new technique of postural evaluation when the measurements are taken by the same tester. Therefore, 3D digitization with magnetic fields could be an interesting alternative to X-Rays for the evaluation of scoliosis. PMID- 9734092 TI - [Polyethylene wear of the Optifix cementless acetabular component after 5 years]. AB - The aim of this study is to review our experience with the Optifix porous-coated cementless acetabular component at an average 5 year follow-up. In a retrospective clinical investigation, the amount and rate of linear wear of the polyethylene of 26 cementless Optifix acetabular components implanted during a primary Hybrid (Spectron femoral component) total hip replacement in 24 patients, were determined by a comparison of the thicknesses of the cup as measured on the initial and most recent follow-up radiographs. The measurements were corrected for magnification error by direct measurement of the diameter of the femoral head on each radiograph according to the technique of Livermore et al. The mean duration of follow-up was 60 months (range, 48 to 73 months). Linear wear averaged 0.53 +/- 0.12 mm (0 to 2.28 mm) overall, with a mean rate of 0.11 +/- 0.02 mm (range 0 to 0.46 mm) per year p < 0.005. No osteolysis of the acetabulum was seen. None of the acetabular components migrated. Osteolysis of the endosteum of the femur was found in six hips. None of the femoral components subsided. In conclusion, wear of the polyethylene in this cup has not been previously reported, and is consistent with results published with other types of acetabular components. PMID- 9734094 TI - [In vitro evaluation of computer-assisted pedicle screw system]. AB - GOAL: The goal of this study is to evaluate influence on the accuracy of pedicle hole positions when a surgeon uses a computer assisted system. STUDY DESIGN: This comparative study was undertaken using dry thoraco-lumbar specimens in order to measure the position of drilled holes with and without computer assistance in the hands of an experienced surgeon as well as with a resident in training. METHODS: Pedicle holes were drilled from D1 to L5 in identical dry thoraco- lumbar specimens. For half of the specimens a computer assisted pedicle screw installation system was used. Holes having been drilled for all specimens, we then measured the maximum distance between the axis of the drilled holes and the pedicle cortices All distances less or equal to 2 mm were classified as safe while all bigger distances were classified as unsafe for pedicle screw installation. RESULTS: Nine specimens were drilled for a total of 306 holes, 170 of them with computer assistance. We have observed a 95% success rate with the computer assisted holes compared to a 62% success rate using conventional methods (p < 0.001). No difference was found between the results of an experienced surgeon and the results of a resident in training while using the computer assistance system. All holes drilled from D12 to L5 were found to be in optimal position, completely within the pedicles. CONCLUSION: These results suggest that this computer assisted pedicle screw installation system could improve the accuracy of pedicle screw placement and enhance the safety of the procedure. Moreover, the computer system could become a valuable teaching tool for the training spine surgeons. PMID- 9734095 TI - [Evaluation of peroperative measurement system in the follow-up of vertebral displacements]. AB - A computer assisted surgery system has been developed to quantify the vertebral displacements induced by the Cotrel-Dubousset instrumentation and to visualize in 3-D the spine at different per-operative steps. This spinal surgery is realized to correct idiopathic scoliosis. Per-operative measurements are obtained by using a magnetic digitizer. Five points per thoracic vertebra and six points per lumbar vertebra are digitized. The method proposed estimates the location of the digitized points on a computer graphics 3-D model of vertebrae by a point-to surface matching algorithm. The 3-D models are constructed before the surgery using multiplanar radiographic reconstruction technique (2 or 3 views) and geometric modeling methods. Computer tools permit the 3-D visualization of the spine peroperatively and the evaluation of clinical indices such as Cobb angles and vertebral rotations. The system includes 3 principal error sources: 2) reconstruction error; 2) digitizer error (digitizer precision) and 3) point-to surface matching error. In order to validate the overall system, measurements have been simulated for 2 vertebrae and 2 digitizers of different precision, taking onto account the reconstruction error. For 5 and 6 digitized points per vertebra, the matching algorithm gives a RMS error of respectively 3.0 and 2.0 mm. PMID- 9734096 TI - [Computer graphic analysis of the three dimensional deformities of scoliotic vertebrae]. AB - GOAL: A computer graphics method that permits the reconstruction, visualization and measure of the vertebral deformities of the scoliotic spine is presented. MATERIALS: Medical imaging techniques utilizing computerized tomography is at the foundation of the reconstruction technique. The studied morphometric parameters are: 1) vertebral body wedging, 2) transverse and spinous process orientation and dimensions and 3) bilateral variation of pedicular dimensions. RESULTS: The reconstructed specimen showed the usefulness of this technique for visualizing and measuring vertebral deformities. Preliminary results seem to be in agreement with the literature concerning the deformities of scoliotic vertebrae. CONCLUSION: This tool will be useful in morphometric investigations for the evaluation of the deformations of scoliotic vertebrae. PMID- 9734097 TI - [Histologic evaluation of bone regeneration in cases of limb lengthening by Ilizarov's technique. An experimental study in the dog]. AB - The histology of bone regeneration in cases of limb lengthening was studied in eight adult dogs. Following an osteotomy of the right fibula, an Orthofix (four dogs) or an Ilizarov external fixator (4 dogs) was installed and an osteotomy of the right tibia performed. Lengthening was started seven days after the surgery at the rate and rhythm of 0.5 mm every 12 hours for three weeks and was then followed by compression of 1 mm. The animals were then sacrificed in pairs 3, 6, 9 and 12 weeks after the start of lengthening. Histological evaluation of regenerate bone was performed using hematoxylin and eosin, trichrome and Von Kossa stain (decalcified). New bone at the site of distraction seemed to be formed mostly through intramembranous, and to a lesser extent, through endochondral ossification as evident by the presence of cartilaginous islands within the distraction gap of old specimens. However, these foci of cartilage cells did not have the appearance of growth plates. From the third week of lengthening, collagen fibers were laid down along the longitudinal axis of distraction. Mineralization of these fibers started at the bone ends and during the following weeks, progressed towards the center of the distraction gap. New bone was formed from both the medullary cavity and the periosteum. PMID- 9734098 TI - [Comparison of prostatic electro-vaporization and transurethral resection in the treatment of benign prostatic hypertrophy]. AB - The objectives of this study was to compare electrovaporization (EVAP) of the prostate to transurethral resection of the prostate (TURP). A prospective evaluation of 60 patients treated for benign prostatic hypertrophy (BPH) was carried out between November 1994 and November 1996. Twenty five patients were treated with TURP and 35 patients with EVAP. All patients had a minimum 12 month follow-up. The results obtained were comparable in terms of surgical procedure time with a bloodless surgical field using EVAP. EVAP was advantageous in reducing the time with indwelling urinary catheter postoperatively and reducing hospital stay. At 12 month follow-up results of flow rates and symptom scores were similar for both TURP and EVAP. Reoperation rate for residual BPH in the EVAP group was higher than for TURP, suggesting that EVAP should probably be limited to smaller sized glands. This study suggests that EVAP is a viable alternative to TURP in selected patients. It requires no specialized equipment and may allow a reduction in catheterization and hospital stay. PMID- 9734099 TI - [Clinical results of peroperative transesophageal echography in peri-valvular leaks of heart prosthesis]. AB - Perivalvular leaks following prosthetic valve replacement are associated with significant morbidity. Management has classically consisted of valve replacement or blind surgical repair. Our study examines the results of intraoperative transesophageal echo-guided repair of perivalvular leaks (ITEGR). Between November 24, 1987 and January 1st, 1996, 23 patients (10 men, 13 women) at the Montreal Heart Institute underwent ITEGR. Ninety percent were NYHA class III-IV preoperatively. Seventy to 85% had significant cardiac insufficiency preoperatively. Eighty-six percent of the leaks were in the mitral valve location, 90% of which were mechanic prosthesis. Eighty-nine percent of patients had hemolysis with an average LDH of 720. Mean bypass time was 125 minutes with a mean clamp time of 77 minutes. Most patients were undergoing a third operation at the time of repair. Operative mortality was 8%, all due to biventricular failure. A mean follow-up of 67 months showed a late death of 10%. Of the 19 survivors, 77% were NYHA class I-II. Overall mortality was 20%. In our institution valve re replacement in similar circumstances was associated with an operative and long term mortality of 7% and 26% respectively. We conclude that intraoperative transesophageal echo-guided repair is an excellent management alternative in patients with perivalvular leaks with decreased late and overall mortality. PMID- 9734100 TI - [Influence of carotid atheroma on the neurologic status after myocardial revascularization]. AB - Surgical management of the carotid disease remains controversial in patients affected with coronary artery atheromatous disease. We report the Montreal Heart Institute experience on the influence of carotid disease on postoperative neurologic events of 501 consecutive patients operated on for coronary revascularization during the period from January 1994 to December 1994. There were 381 men and 114 women averaging 62 +/- 9 years old. Major risk factors were high blood pressure (35%), and smoking habit (48%). Fifty-nine patients presented clinical signs of carotid atheromatosis and among them 21 had significant carotid stenosis (> 80% decrease of cross sectional area). During surgery, the mean duration of extracorporeal circulation (ECC) was 76 +/- 31 minutes and the mean perfusion pressure (MPP) was 70 +/- 11 mmHg. The use of inotropic drugs was mandatory in 26% of the cases and the mean arterial lactate (AL) dosage during ECG was 3.07 +/- 1.35 mM/L. During the perioperative period, 13 (2.5%) patients sustained neurologic disturbances of which 5 (1%) were lateralized. Among them, 8 completely recovered whereas 3 of the 5 with permanent damage died. None of the patients with preoperative stigmata of carotid disease experienced lateralized neurologic deficit. Multivariate regression analysis identified the use of vasopressor drugs and perioperative increase of AL as predictive factors. We conclude that in our series, the incidence of neurologic complications was low. The presence of carotid atheromatosis did not increase the postsurgical risk of cerebrovascular accident, however, the increased incidence of neurologic events associated with inotropic drugs and increased AL suggests a direct link with a systemic oxygen debt. Consequently, we do no recommend concurrent prophylactic surgery during coronary artery revascularization. PMID- 9734101 TI - [Effects of chronic administration of captopril on vascular reactivity of the rat aorta]. AB - The vascular endothelial function is altered in certain vascular pathology such as arterial hypertension. However, the endothelial effect of the pharmacologic treatment of this pathology with angiotensin converting enzyme (ACE) inhibitor is poorly understood. To evaluate the effects of long-term treatment of captopril on the rat aortic vascular reactivity, 2 groups of spontaneously hypertensive rats (SHR) (n = 6) were studied for 12 weeks: the first group was treated with captopril (CAP) (2 g/l of water) while group 2 (OCAP) received no treatment. A third group without hypertension was considered as control group (CTL). At the end of the experiments, isolated aortic rings were studied in organ chambers for endothelial and smooth muscle vascular reactivity. The endothelial-dependent relaxations (EDR) to cumulative doses of acetylcholine, histamine or adenosine diphosphate were significantly decreased in the aortic segments of CTL compared to CAP (p < 0.05). Aortic segments from OCAP group demonstrated intermediate EDR responses between CAP and CTL groups. Smooth muscle relaxation to sodium nitroprusside was comparable among the 3 groups. Maximal smooth muscle contraction to progressive doses of norepinephrine was significantly higher in the CTL group compared to the hypertensive groups. CONCLUSION: Spontaneously hypertensive rats have an increased basal vascular tone. The increased EDR observed with hypertension partially compensates this hypercontractility. Chronic hypertension treatment with captopril partially normalizes EDR responses in the rat aorta suggesting the lost of the endothelial compensatory mechanism by ACE inhibitors. PMID- 9734102 TI - [Revascularization of the circumflex artery with mechanical stabilization: initial experience]. AB - OBJECTIVES: To evaluate the short-term result of the coronary artery revascularization without cardiopulmonary bypass for triple vessel disease, including the circumflex territory performed on the stabilized beating heart. METHODS: Prospective study conducted on the first 35 consecutive patients with triple vessel disease operated upon without cardiopulmonary bypass by a single surgeon (RC) at the Montreal Heart Institute between October 1996 and March 1997. RESULTS: Mean age of patients was 64 +/-1.6 years and the majority were men (30). Most common risk factors were hypercholesterolemia (65%) and familial history (55%) of ischemic heart disease. Main surgical indication was unstable angina (74%) and mean preoperative left ventricular ejection fraction was 53 +/- 3%. Hundred and twelve bypass were constructed averaging 3.2 +/- 0.1 grafts/patients of which 39 were made on branches of the circumflex artery. Average ischemic time was 34.17 +/- 2.17 minutes. The internal thoracic artery, saphenous vein, and radial artery were used as a vascular conduit in 44, 67, and 1 occasions respectively. There was one operative mortality, and one non Q perioperative myocardial infarction (CK-MB: 89 U/L). No patient required aortic counterpulsation balloon assistance. The average postoperative CK-MB (U/L) were 12.2 +/- 1.9, 15.2 +/- 3.2, and 10.3 +/- 1.7 at 1, 24 and 48 hours respectively. During the post-operative period 26% (9) of the patients presented atrial fibrillation, 6.5% (2) early reexploration for bleeding, and 63% (22) did not require transfusion. Average stay in hospital was 6.1 +/- 45 days. Coronary grafts were angiographically assessed in the first 10 patients and at the postmortem exam in one and displayed a 100% patency with 93.5% (29/31) adequate runoff. CONCLUSION: Triple vessel coronary artery disease revascularization is feasible on the beating heart without cardiopulmonary bypass with excellent short term clinical and angiographic results. PMID- 9734103 TI - [Cyclosporine A prevents ischemia-reperfusion induced myocardial dysfunction in the isolated heart of the rat]. AB - Cyclosporin A (CyA) has been shown to prevent mitochondrial injury following ischemia-reperfusion injury. Therefore, the present study was designed to investigate the effect of CyA on ischemia-reperfusion injury in the isolated rat heart preparation. Hearts from Sprague-Dawley rats were perfused in the Langerdorffmode at constant pressure (80 cm H2O) and paced (270 beats/min). After equilibration, hearts were treated with CyA (10(-5) mol/l) (n = 8) or its vehicle, cremophor (Cr) (n = 8) for 10 minutes before exposure to 30 minutes of global ischemia and 60 minutes of reperfusion. Hemodynamic variable vascular reactivity, and oxygen consumption (MVO2) were tested at baseline and at selected points during reperfusion Hemodynamic variables were significantly improved in the CyA group. The maximal mean percentage of preservation for left ventricular developed pressure (PDVG) was -14.9 +/- 10.7% and +31.5 +/- 23.6% respectively for Cr and CyA group (p<0.05) The maximal mean percentage of preservation for dp/dt was -11.7 +/- 11.4% and +28.3 +/- 29.9% respectively for Cr and CyA group (p < 0.05): the compliance, -dP/dt was also preserved, maximal mean preservation was -25.9 +/- 9.2% and +53.1 + 30.1% respectively in Cr and CyA group (p < 0.01). Oxygen debt was decreased at 30 minutes of reperfusion in the CyA-treated hearts: 0.06 x 10(-2) +/- 0.23 x 10(-2) cc/min/g compared to Cr-treated hearts: 0.61 x 10(-2) +/- 0.37 x 10(-2) cc/min/g (p = 0.05). The coronary endothelial dependent and independent responses were similarly decreased in both groups during reperfusion. Thus, in the isolated rat heart preparation, CyA preserves myocardial function (hemodynamic variables) and oxygen consumption without affecting the coronary vascular function during ischemia-reperfusion injury. PMID- 9734104 TI - [Central dopaminergic D1 agonists and treatment of Parkinson disease]. AB - Many theories about dopaminergic function in Parkinson's disease are based upon the effects of the D2 receptor. Standard treatments mostly involve dopaminergic D2 agonists. However, the recent development of dopaminergic D1 agonists should help to clarify the role of the D1 receptor in the treatment of Parkinson's disease. The authors review the physiopathological, behavioural and therapeutic data on D1 agonists administered in animal models and in patients. PMID- 9734105 TI - [Multifactor analysis: method and application for the selection of molecules in cardiovascular diseases]. AB - Among the molecules under development or registration proposes for licensing among the pharmaceutical firms, the choice of one molecule is often difficult. The methods of decision analysis can allowed the formalization of this choice. Multicriterion analyses (ELECTRE 1 and 2) have the capacity to integrate the qualitative criteria and to incorporate concordance and discordance indicators in the judgement. The objective of this study is to present the bases of the multicriterion method and to give an example of application in the clinical cardiovascular domain to select one anti-hypertensive molecule among a preselection of 9, by using 12 differently weighted judgment criteria. Five molecules were selected by the ELECTRE 1 method and only one by the ELECTRE2 method. PMID- 9734106 TI - [Numeric case report: realization and practical utilization in cardiovascular research]. AB - The reliability of trials in clinical research is principally conditioned by the quality of the conception, the realization and the data recording in the hypothesis of pertinent trials with an adequate methodology. The author describes the application in the cardiovascular domain, the realization and the use of numeric Case Report Forms DCRF working with a 'notebook' microcomputer and easy to use software which allows the conduct of quality clinical trials in accordance with good clinical practice. The numeric CRF is a tool to monitor in real time the data from trials (ongoing data trials) and ensures the continuity from the investigator site to the statistical department. An example of realization of a phase III multicentre clinical study is presented to evaluate the efficacy of two antihypertensive calcium-channel blockers. PMID- 9734107 TI - [Cost evaluation of therapeutic management of patients with chronic hepatitis C]. AB - Hepatitis C now poses an important problem of public health in France. The objective of this study was to evaluate the cost of care of 55 patients who were carriers of hepatitis C and treated with interferon alpha. The economic analysis included the blood testing costs, pharmaceutical costs and hospitalization costs. A questionnaire permitted the impact of the treatment on the socio-professional life of each individual to be assessed. The global cost amounted to 1155 359 francs spread among hospitalization costs (55 pour cent), pharmaceutical costs (35.8 pour cent) and blood testing costs (9.2 pour cent). Of 36 actively working patients being treated, 10 modified their activities and 4 of these 10 restarted their initial activities after stopping treatment. This evaluation has shown the disparities which exist in the clinical follow up of patients treated for chronic hepatitis C. It is thus imperative to institute a systematic screening of patients being transfused to permit an early diagnosis, to establish a rigorous standardization of blood testing, to visualise day care treatment and above all to establish a consensus of global care of these patients. PMID- 9734108 TI - [A survey on the practice of drug prescription in hospitals. Concepts and methodology]. AB - A clinical practice survey, if patient focused, identifies and analyses complexity of drug use. It is not a clinical audit whose aim is to compare observed practice with predetermined criteria. A clinical practice survey is useful when a problem exists in a low evidence based clinical area or to elaborate clinical guidelines. Implementation is based on epidemiological methodology and project management and needs a framework determined by a facilitator (trained in quality improvement methods and expert in the clinical area concerned), planning (the more higher the number of wards, the higher the complexity) and the project guidelines must be respected. Since lack of physician involvement is a drawback, support provided by the quality centre (if it exists) must be limited to technical aspects. It is important to appreciate the level of quality culture of the organization and the project must be approved by the hospital manager. PMID- 9734109 TI - [Study of the quality of prescribing drugs in hospitals]. AB - The quality of prescription writing has been evaluated in respect of standards of good practice. Among the 44 medical departments of a university hospital, 39 agreed to participate in the study. A sample of 30 patients from each participating department was randomized out of one year's hospital stays; 790 patient records were relevant and have been analysed. The patient's identification was completed in 39.1 +/ 3.4 per cent. THe prescriptor was properly identified by name and signature in 7.2 +/ 1.8 per cent of prescriptions studied. Only 8.9 +/ 2.1 per cent of prescriptions contained the required information for each medication. Results of this study have been sent to all hospital physicians with recommendations for good prescription practice. This evaluation constitutes the first stage of a quality process based on the awareness and information of concerned actors and follow up of specific indicators. PMID- 9734110 TI - [Analysis of prescriptions for psychotropic drugs in a psychiatric hospital]. AB - A study on the prescription of psychotropic drugs was carried out at St-Egreve Psychiatric Hospital (Isere,--France) based on in-patient prescriptions (adults only). Analysis of the 200 prescriptions issued by 26 different practitioners found an average of 3.3 psychotropic drugs per patient (including 1.7 neuroleptics). A combination of neuroleptics was very frequent (63 pour cent of the schizophrenics receiving at least 2 neuroleptics). Half of the patients treated with neuroleptics received antiparkinsonian drugs. The other psychotropic medications often combined with neuroleptics, were anxiolytics (50 per cent), hypnotics (45 per cent), antidepressants (28 per cent) and normothymics (25 per cent). Among the antidepressants, the SSRI were the most commonly prescribed, even above tricyclics, reflecting their better clinical tolerance. The significant differences in prescribing practice between the services prompted the creation of a working group comprising practitioners and pharmacists to study the prescription of psychotropic drugs and compare this with the literature. PMID- 9734111 TI - [Proposal of guidelines for the set-up of pharmaco-epidemiologic studies in drug surveillance]. AB - In the light of recent experiences and anticipating an increase in similar requests in the future, it seemed very interesting to a drug safety executives' group from the pharmaceutical industry to propose guidelines for the set-up and follow-up of pharmacoepidemiological studies requested by Health Authorities for the assessment of drug risk. The scope of these guidelines is to establish the responsibility of the teams and structures involved in the study, to define the necessary stages set-up, and to determine the rules in order to ensure its smooth running from the drafting of the protocole to the final use of the data. PMID- 9734112 TI - [Esophageal involvement after tetracycline ingestion]. AB - Between 1985 and 1992, 81 spontaneous oesophageal injuries associated with tetracycline were notified to the French Regional Pharmacovigilance Centres. The side effects were oesophageal ulcers (79 per cent), esophagitis (11 per cent) and dysphagia (10 per cent). Esophagitis and dysphagia appeared sooner (4 days) than the ulcers (15 days). The mean age of the patients was 29 +/ 13 years and 73 per cent were women. In 92 per cent of cases, the recommendations for administration were not observed (medication taken at bedtime with not enough or without water). With 96 per cent of patients, doxycycline was the tetracycline in question; this prevalence could be explained by its irritant and cytotoxic properties. The oesophageal injuries were 22 times more frequent with capsules than with tablets, because of their easier adhesion to the oesophageal surface. Oesophageal injuries are potentially serious and must be avoided by clear information to patients and prescribers on tetracycline administration; consumption in the middle of a meal with an adequate quantity of water and never less than one hour before bedtime. PMID- 9734113 TI - [Thrombocytopenia due to heparin therapy. Use of danaparoid (Orgaran), 13 monocentric cases under authorization of temporary prescription (ATU)]. AB - Since September 1994, danaparoid (Orgaran), a heparinoid, has been used in our centre to treat patients with thrombocytopenia occurring during heparin therapy and who need continuing antithrombotic therapy. We carried out a retrospective study using clinical and biological data on the first 13 consecutive patients treated with danaparoid (for 1 to 18 consecutive days). The platelet count returned to normal for ten patients, but one patient died having contracted a severe sepsis and bleeding occurred in one patient with acute renal failure. In the three other cases, the diagnosis of heparin induced thrombocytopenia (HIT) was in retrospect unlikely and the death of these patients was related to severe underlying diseases which were held responsible for thrombocytopenia. We confirm that danaparoid appears to be an effective, well-tolerated substitute for heparin in HIT patients. The French regulation Temporary Authorization for Prescribing Medicines allowed the prompt use of this as yet unmarketed drug and collection of reliable and pertinent data. PMID- 9734114 TI - [Risk factors of adverse effects of angiotensin-converting enzyme inhibitors. Apropos of 30,072 patients treated by trandolapril]. AB - SUMMARY: The aim of the present study was to investigate the incidence of adverse effects and the prognostic value of various risk factors in a large population of unselected hypertensive patients treated with the ACE inhibitor trandolapril. Among the 30 072 patients investigated in this post marketing retrospective study, 1813 patients (6.0 per cent) reported an adverse effect. The five most frequent side effects were coughing (3.1 per cent), dizziness (0.7 per cent), headache (0.6 per cent) asthenia (0.5 per cent) and nausea (0.3 per cent). Intolerance risk factors for trandolapril were researched using both univariate and multivariate analysis. In the univariate analysis, a prior intolerance of an ACE inhibitor and female gender were strongly correlated with either overall intolerance or coughing. The most relevant variables for the occurrence of adverse effects, listed according to their entry order in the multivariate analysis, were: prior intolerance of ACE inhibitors (OR: 4.19, 95 per cent CI: 3.66-4.78), female gender (OR: 1.46, 95 per cent CI: 1.31-1.63), prior intolerance of other antihpertensive agents (OR: 1.27, 95 per cent CI: 1.14 1.41), smoking (OR: 0.76, 95 per cent CI: 0.66-0.87) and combination with a beta blocker (OR: 1.31, 95 per cent CI: 1.08-1.58). A prior intolerance of an ACE inhibitor appears to be a very strong predictor of coughing (OR: 6.14, 95 per cent CI: 5.24-7.19). The following variables, namely female gender (OR: 1.61, 95 per cent CI: 1.40-1.85), age 60-80 (OR: 1.25, 95 per cent CI: 1.09-1.44) and prior intolerance of other antihypertensive agents (OR: 1.20, 95 per cent CI: 1.03-2.39) appear less significant. PMID- 9734116 TI - Fluvoxamine interaction with fluindione: a case report. PMID- 9734115 TI - Dextromethorphan poisoning in an adolescent with genetic cytochrome P450 CYP2D6 deficiency. PMID- 9734117 TI - [Granulomatous hepatitis and ticlopidine]. PMID- 9734118 TI - [Acid pump inhibitors and pregnancy: apropos of 24 women with known development]. PMID- 9734119 TI - [Is hemodialysis a risk factor for tendinopathies due to fluoroquinolones?]. PMID- 9734121 TI - [Current status of echoguided transrectal biopsy of the prostate gland]. PMID- 9734120 TI - Lipodystrophia with protease inhibitors in HIV patients. PMID- 9734122 TI - [Initial aspects of a program for in vitro transfection of cytokine genes into LNCap tumor cells]. AB - OBJECTIVES: To characterize the conditions and kinetics for in vitro growth of tumoral cells LNCaP. METHODS: Determination of proliferation curves and time to cell duplication through spectrophotometry and manual cell counts in haemocytometer with and without foetal calf serum (FCS) challenge. RESULTS: Data from the spectrophotometry and manual cell counts were correlated by mean of the equation through the number of cells = absorbance at 595 nm x 227,530-14,160. The correlation coefficient r was 0.99. Cell proliferation was higher in the presence of FCS (p > 0.001), the final culture density being 5.4 times higher than the baseline. Similarly, time to duplication in the presence of FCS was 36.4 hours. CONCLUSIONS: This experiment allowed to establish the local conditions for growth of LNCaP cells. In contrast to the behaviour exhibited by other malignant cells in culture, LNCaP cells grow slowly and require the stimulus provided by FCS. Both aspect are of the highest significance when adopting the pre-established timetables and logistic to the actual laboratory operation. PMID- 9734123 TI - [Shared care in BPH. First national experience]. AB - The high prevalence of Benign Prostate Hyperplasia and the increased demand for care of this condition, should compel us to plan for shared care models in parallel to Primary Care, in the way it has happened with entities such as HBP and Diabetes. The set of measurements to be adopted when sharing services with primary care is known as "shared care". This paper presents the first national experience of "shared care" with primary care in BPH. The project has consisted in a series of steps to increase awareness, train and make available for family physicians, a clinical practice guide defining the criteria for initial evaluation, medical treatment and referral of patients to Urology surgeries, including with the referral document the appropriate diagnostic tests. A Quality Commission has been created to study the level of compliance of the documentation used for referral to the specialist and the clinical histories of patients treated in primary care. The results obtained are significant and most studies carried out fulfill the requirements in 60% cases, which has allowed to reduce overcrowding in the Urology outpatient offices (4200 surgery visits saved/year in our environment), has provided easy access of patients to adequate diagnosis and treatment, as well as significant financial savings (30 million pesetas/year). In short "shared care" is a reality in our environment that allows a more effective, fast medical assistance and improved access to specialist care by reducing the demand of specialized surgery hours. PMID- 9734124 TI - [Flowmetry analysis in patients undergoing transurethral resection of the prostate for BPH]. AB - Transurethral resection (TUR) is the most frequent surgical treatment for symptomatic benign prostate hyperplasia (BPH). Prostate size is a significant factor for choosing TUR versus prostate adenomectomy. Analysis of flowmetry results obtained with TUR in 203 patients, based on weight of resected prostate tissue. Flowmetry was performed prior to TUR and prostate size was estimated with transabdominal ultrasound. Prostate tissue was weighed after TUR and a new flowmetry was performed 6 months after treatment. Mean weight of resected tissue was 31.34 g. When all flowmetry parameters analyzed pre-and post TURs were compare, there were significant differences (p < 0.001). Increased maximum flow (Qmax) and increased mean flow (Qmed) occurred in 91.7% and 96.31% patients, respectively. Mean increase of post-surgical Qmax was significantly higher (p > 0.01) in patients with Qmax prior to surgery lower than 8 ml/s. No significant correlation was demonstrated between prostate volume measured by ultrasound or resected prostate tissue and increased post-surgical Qmax. TUR improves flowmetry parameters, mainly in patients with pre-surgical Qmax lower than 8 mL/s. Extensive prostate resection does not appear to improve the flowmetry results obtained with a sufficient functional TUR. PMID- 9734126 TI - [Pathology of the peritoneo-vaginal complex in the child. Communicating hydrocele, cord cyst, simple hydrocele]. AB - The abnormal persistence of the patent processus vaginalis determines the appearance of four types of pathology, depending on the grade and sort of communication: communicating hydrocele, hydrocele of cord, scrotal hydrocele and intrafunicular hernia. We have revised our casuistry of children with patent processus vaginalis pathology for the two last years (1995-1996), and we have found 75 communicating hydrocele cases, 5 hydrocele of cord and 16 scrotal hydrocele cases, on children between 1 month and 13 years old. The diagnostic was done after physical exploration with transillumination and inguino-scrotal ultrasound. Initially, conservative treatment was followed, which was enough for 58 patients (60.4%). In the 38 cases (39.6%) in which there were no improvement, surgical treatment via inguinal was carried out, with good results in nearly all cases. As a conclusion, we can assert that ultrasound is an excellent diagnostical method for patent processus vaginalis pathology and conservative treatment must have priority upon surgery, since a great number of spontaneous resolutions are observed, most of all on children aged less than two years old. PMID- 9734127 TI - [Extraperitoneal anterior approach to renal vessels in radical surgery of renal cancer]. AB - The authors present a modification of Droller approach to the renal vessels prior to any Kidney manipulation, even in fat patients. The anatomical situation of the hilium elements is on the basis of their rationale. The ascending ureter dissection allows the blunt advance of the middle finger by the posterior way until the arterial beat. Then pushing up the artery, its identification, by the anterior way, is facilitated. With an initial series of 7 cases the results show no ileus complication and shorter hospital stay. This extraperitoneal approach enables to keep oncologic surgical rules in selected cases of renal cell carcinoma. PMID- 9734125 TI - [Permanent endoprostatic prostheses in patient with obstruction caused by benign hyperplasia of the prostate]. AB - Presentation of the results obtained using the intraprostatic prosthesis UroLume in 78 patients wit BPH obstruction, 69 of which presented high surgical risk (ASA IV). Mean age was 79.8 years (r: 62-93). All patients carried urethral catheters, except 4 (5.1%) who had a provisional metal coil that required replacement. Prosthesis were implanted successfully in 72 cases (93.3%). The most significant exclusion criterion was an excessive length of prostate urethra. Mean follow-up was 15.3 months (r: 3-38). Mean maximum flow at 1 year after implant was 12.7 mL/sec; mean symptoms score (I-PSS score) was 6.2 points and in most prosthesis, epithelization had taken place. Three patients required implant of another prosthesis, either during the same surgical procedure (1 case) or later due to retention or dysuria (2 cases). Due to acute urine retention (AUR) during the immediate postoperative, resection of the middle lobe was performed in one case while a second case required late resection of intraluminal hyperplastic tissue. Three patients (4.1%) had haematuria that forced hospital admission some months after the implant, and three cases (4.1%) required removal of the prosthesis; at patient's request (1 case), due to calcification (1 case) and for stress incontinence (1 case). After a follow-up of over three years, it can be concluded that the UroLume prosthesis is an effective alternative to TUR in patients at high surgical risk. PMID- 9734128 TI - [Renal adenocarcinoma in children]. AB - Contribution of two cases of renal adenocarcinoma in pediatric patients. Comments on the most updated and relevant aspects of epidemiology, clinical manifestations, diagnosis, treatment, prognosis and follow-up. Comparison to nephroblastoma and renal adenocarcinoma of the adult. PMID- 9734129 TI - [Paratesticular sarcoma: apropos of a case with a 19-year evolution]. AB - Presentation of one case report of paratesticular sarcoma in a 51-year old patient undergoing surgical treatment and adjuvant systemic radio- and chemo therapy with 19 years follow-up, that evolved with local relapse of the disease but no distant spread. Analysis of the difficulties of the pathoanatomical diagnosis, currently improved by immunohistochemical methods and the value of the adjuvant treatment. PMID- 9734130 TI - [Mucinous adenocarcinoma of the urachus synchronic with colorectal adenocarcinoma. Value of immunohistochemistry in the differential diagnosis]. AB - The coincidence of an urachal adenocarcinoma with another similar tumour in other location makes necessary to separate a true primary from a metastatic adenocarcinoma. We report the case of a 66-years-old-man with an urachal mucinous adenocarcinoma and two colonic adenocarcinoma excised in the same surgical act, showing both macro and microscopic studies together with immunohistochemical techniques, that were useful to differentiate the origin of both neoplasms. Among them, the antibody to keratin 7, a cytoplasmic epithelial protein, was positive in the urachal and negative in the colonic tumour. PMID- 9734131 TI - [Severe hematuria caused by radiation cystitis. Selective percutaneous embolization as an alternative therapy]. AB - Post-radiotherapy cystitis is a high morbidity entity that involves difficult-to treat haematurias and sometimes results in death. Percutaneous arterial embolization is an accepted approach for haemorrhages of pelvic origin. Selective, percutaneous embolization of both vesical arteries could be an alternative treatment to control severe haematurias caused by radiation. Presentation of our experience in two cases and proposal of this approach as a valid option in post-radiotherapy haemorrhagic cystitis. PMID- 9734132 TI - [Retroperitoneal benign fibrous histiocytoma. Presentation of a case]. AB - Presentation of one case of an extraperitoneally located Benign Fibrous Histiocytoma in a 59-year old male. Retroperitoneal tumours are extremely rare neoplasias, malignant in about 85% cases 35% of which are sarcomas. The benign fibrous histiocytoma is most often located in subcutaneous tissues, a deeper location being exceptional. They are usually clinically silent entities until size causes a shift or compression of other organs which cause the clinical signs and symptoms. In many cases, the neoplasia is detected by ultrasound. Computerized Axial Tomography is the technique that provides more data for the establishment of diagnosis, although for many authors Nuclear Magnetic Resonance can be more defining. The primary therapy is surgical removal, but relapses can occur if full exeresis is not achieved. PMID- 9734133 TI - [Right varicocele as first manifestation of situs inversus]. AB - Traditionally, right varicoceles have been considered a rare entity nearly always secondary to a neoplastic or retroperitoneal disease. There are other possible etiologies for right varicoceles such as the venous disease of the large veins and visceral malposition syndromes. This paper presents one case report of right varicocele as the only clinical sign of situs inversus. Revision of the literature and nomenclature of these syndromes. PMID- 9734134 TI - [Posttraumatic high-flow priapism in prepubertal age]. AB - High flow priapism is an infrequent entity, generally following traumatic injuries in the genito-perineal area. Anamnesis, cavernous bodies blood gasometry and Doppler are the basic diagnostic tools for these condition. Therapeutical management is considerably different from that used for low flow venous priapism. Selective arteriography of the internal pudendal artery allows to locate the arterial lesion and, at the same time, to perform supraselective embolization of the lacerated cavernous artery which is currently considered the choice treatment. This paper presents the case report of a ten-year old patient successfully resolved through application of angioradiologic procedures. PMID- 9734135 TI - [Acute epididymitis in a 3-month-old infant: an infrequent presentation]. AB - The acute Epididymitis is a very uncommon disease in children under 3 years old. We present the clinic case in a suckling child of 3 months old with onset of an acute scrotum, being necessary the surgical exploration to define the exact diagnosis of acute Epididymitis. We make a revision on world medical references and found out a very low incidence in children under 1 year old (0.7-3%); so it is considered of interest to publish it with the double finality to be taken into account in the presence of an acute scrotum and to contribute with a new case of this unusual pathology in babies under 1 year old. PMID- 9734136 TI - [Rare origin of a retroperitoneal mass: Castleman's disease]. AB - Castleman's disease (angiofollicular hyperplasia of the lymphatic nodes) can exceptionally appear as a retroperitoneal mass of difficult differential diagnosis relative to other malignant retroperitoneal masses. Because of its rarity, one case report of a retroperitoneal mass with histologic study corresponding to Castleman's disease is contributed. A revision of the different histologic varieties of Castleman's disease, specific treatment and prognosis is included. PMID- 9734137 TI - [New chemotherapy regimens for advanced bladder cancer]. PMID- 9734138 TI - ["Internal" echography or echography for the internist]. PMID- 9734139 TI - Exposing the mast cell: its novel integrated role in allergy. AB - New discoveries have suggested that the mast cell has the potential to regulate allergic inflammation by inducing IgE synthesis from B cells. Under allergic inflammatory conditions, "primed" mast cells appear to express higher levels of the high affinity receptor for IgE and the ligand for the surface antigen CD40, involved in T/B cell interactions leading to immunoglobulin production, as well as Th2-type cytokines, IL-4 and IL-13. The critical role of these cells in the induction of IgE synthesis is supported by the findings that anti-ligand for the surface antigen CD40, anti-IL-4, and anti-IL-13 monoclonal antibodies inhibit IgE production. Mast cells also have the potential to function as antigen presenting cells with the ability to shift T cells into Th2 subtypes. These recent findings suggest that mast cells can modulate important regulatory functions of the allergic response by acting directly on B cells and inducing IgE production. PMID- 9734140 TI - Clinical application of fibrinolysis laboratory tests: a review. AB - The aim of this paper is to review fibrinolysis laboratory tests of potential clinical usefulness. Since the activation of the fibrinolytic system may be responsible for several relevant pathological scenarios, it is crucial to know whether and to what extent fibrinolysis laboratory tests are useful for the diagnosis and therapy of individual patients. The plasma fibrinolytic system may be altered by deficiencies and/or abnormalities of some of its components. Few doubts remain concerning the possible role of fibrinolysis alterations in the pathophysiology of some hemorrhagic and thrombotic disorders. In hemorrhagic patients, laboratory tests may demonstrate the existence of increased plasmin activity and the presence of specific congenital defects leading to primary hyperfibrinolysis. Alterations of the fibrinolytic system should be looked for only in selected patients who have a history of venous thrombosis and negative results of initial screening tests. PMID- 9734141 TI - Endothelins: an overview of recent achievements. AB - The aim of this study is to review the recent developments in research on the pathophysiological role of endothelins in the reproductive system and lungs, as well as in systemic disorders such as sepsis, acute respiratory distress syndrome and multiple organ failure. The international literature of the last 9 years, as available on MEDLINE, was duly scanned. Only relevant, original and update studies were selected for the purpose of this survey. The results of the most significant studies in humans are summarized. The potential roles of endothelins in systemic diseases are currently under investigation. A possible role of these peptides is suggested in the pathophysiology of heart failure and in the mechanism of post-ischemic renal failure. Studies have been conducted also on other systems, namely the reproductive system, the endocrine system and the lungs as well as on a number of tissue disorders such as sepsis, acute respiratory distress syndrome multiple organ failure, preeclampsia and related abnormalities. Full understanding of the role and action of this group of regulatory mediators in tissues should enable complete insight into the pathophysiological mechanisms of localized diseases as well as of multiple organ diseases. As a result, adequate therapy may be established. PMID- 9734143 TI - [Neuroleptic malignant syndrome: a neurologic pathology of great interest for the internist]. AB - Two cases of neuroleptic malignant syndrome are reported. Although affecting chiefly psychiatric patients, this rare, polymorphic disease should be well-known to internists, general practitioners, and emergency staff because of its high risk of fatality. Our patients, who presented without some of the main symptoms, fell in the category of incomplete variants of the syndrome, the most difficult to recognize. In both patients, computerized tomography carried out at the outset of symptoms, evidenced signs of cerebral edema, a datum never before reported. The symptomatic polymorphism, pathogenesis, differential diagnosis, and therapy of this disease are illustrated and discussed. PMID- 9734142 TI - New advances in the pathogenesis and therapy of bronchial asthma. AB - At present, general consensus exits on the inflammatory nature of bronchial asthma. Advances in the knowledge of pathogenetic mechanisms of bronchial asthma are certainly to be ascribed to the use of immunobiological, molecular and genetic approaches for studying experimental models and human bronchial asthma. The histopathological hallmark of bronchial asthma consists in the constant presence, at the level of bronchial mucosa, even in the mild and intermittent syndromes, of epithelial lesions, thickening of basement membrane, and inflammatory infiltration consisting of activated eosinophils, Th2 lymphocytes and degranulated mast cells. Cellular and molecular alterations responsible for this typical bronchial inflammation and the subsequent patho-physiological and clinical characteristics of bronchial asthma have been identified. It has been shown that interleukin-4 is the principal cytokine necessary for Th2 lymphocyte differentiation and expansion and IgE production. Interleukin-5 acts specifically on eosinophil activation, terminal differentiation, and survival. As only eosinophils primed by interleukin-5 as well as by interleukin-3 and granulocyte macrophage-colony stimulating factor can express all the membrane receptors and integrins, they become able to bind to adhesion molecules overexpressed on the mucosal microvascular endothelial cells and of being selectively chemoattracted by a particular subgroup of beta-chemokines, which bind to their membrane CC receptor-3. The amplification and maintenance of airway inflammation and the clinical exacerbations and chronicity of bronchial asthma depend on complex mechanisms. Tumor necrosis factor-alpha and interleukin-1 beta play a fundamental role in the induction of multiple cellular and molecular interactions. Molecular approaches are now being used to define genetic predisposition. Candidate genes have been located on different chromosomes. In addition to bronchodilator medication to relieve symptoms, the most important purpose of asthma therapy must be based on anti-inflammatory agents to reduce airway inflammation and airway hyperreactivity. Corticosteroids appear able to interfere with almost all components of inflammation. New inhaled steroids have radically modified therapeutic principles of bronchial asthma. Even cromones and anti-leukotrienes have been recommended as anti-inflammatory agents. Some developing therapeutic strategies involve use of anti-IgE antibodies, interleukin-4 and interleukin-5 inhibitors and chemokine receptor antagonists. PMID- 9734144 TI - [Castleman's disease with isolated renal location: clinical case]. AB - In a review of the literature the authors delineate the present nosographic and descriptive characteristics of Castleman's disease. They then report the case of an adult woman who came to their attention because of persistent, low-grade fever, sweating, malaise and polyarthralgia. Laboratory data evidenced increased acute-phase reactants, polyclonal hypergammaglobulinemia, and anemia due to "chronic disease". Diagnostic imaging documented a right renal mass. A nephrectomy was performed. Histopathological studies confirmed hyaline-vascular type Castleman's disease with monoclonal B-cell lymphoproliferation. The clinical and laboratory anomalies regressed after surgery and continue to be absent after 1 year of follow-up. The authors conclude their presentation by pointing out the peculiarities of this case that do not correspond with the traditional distinctive features of the disease. PMID- 9734145 TI - [Clinical reflexions on a case of primary achalasia of the esophagus diagnosed late]. AB - We describe an unusual case of achalasia. The patient, a 33-year-old woman, presented with a clinical history of esophageal disease verified by gastroscopy. The diagnosis of hysterical anorexia that had been made some years previously did not correspond with the nosological classifications (DSM III-R, DSM IV). This case underscores the importance of the correct use of clinical methodology, particularly when conclusive diagnosis is essential for successful treatment. PMID- 9734146 TI - [Gastrointestinal stromal tumor as the cause of intestinal hemorrhage: description of a clinical case]. AB - The term gastrointestinal stromal tumor describes a heterogeneous group of tumors of mesenchymal origin with particular histologic features. Their classification has recently been made possible thanks to numerous immunohistochemical and ultrastructural studies. We report the case of a patient who came to our attention because of serious anemia due to a gastrointestinal stromal tumor located between the second and third portion of the duodenum. This pathology, although not frequent, should be considered in the differential diagnosis of gastrointestinal bleeding. In these cases it is important to perform careful endoscopic inspection to the third portion of th duodenum, even when previous tracts evidence lesions that could be responsible for the patient's symptoms. PMID- 9734147 TI - ["Puerto Rican medicine at the threshold of a new century"]. PMID- 9734148 TI - [Quo vadis. Where is medical education going at the end of the Twentieth Century?]. PMID- 9734149 TI - [Our opinion on "Medicine among the indians". Bulletin editorial Medical Association of Puerto Rico--October 1904]. PMID- 9734150 TI - Epidemiological trends of melanoma in Puerto Rico from 1975-1991. AB - The purpose of this study was to determine the crude and age-adjusted incidence rates of melanoma for residents of Puerto Rico from 1975 to 1991. This is part of an ongoing NASA study aimed at estimating whether melanoma and cataracts have increased in Puerto Rico since 1978 because of potential stratospheric ozone depletion and increased ultraviolet-B (UV-B) radiation. Calculating the percent change from their lowest values in 1978 to 1991, the age-adjusted incidence rate of melanoma increased 528% for males and 200% for females in 13 years. PMID- 9734151 TI - CCR5 chemokine receptor genotype frequencies among Puerto Rican HIV-1 seropositive individuals. AB - Some individuals remain uninfected by human immunodeficiency virus type 1 (HIV 1), despite multiple sexual contacts with subjects with confirmed HIV-1 infection. Several studies have confirmed that individuals who are homozygous for a 32 base pair (bp) deletion mutation in the chemokine receptor gene CCR5, designated as delta 32/ delta 32, are protected against HIV-1 infection. Heterozygotes of the same chemokine receptor deletion mutation are, however, not protected from acquiring HIV-1 infection but seemingly have slower progression to acquired immunodeficiency syndromes (AIDS). Genotype frequencies of the delta 32 CCR5 mutation vary markedly among different ethnic groups; heterozygosity is found in approximately 15% of Caucasians, about 5-7% of Hispanics and African Americans and 1% or less of Asians. The ethnic background of Puerto Ricans is highly complex and usually includes admixture of Caucasian, Caribbean Indian and African traits to a varying extent. This study was conducted to examine the frequencies of the delta 32 CCR5 mutation among Puerto Ricans who are infected with HIV-1. Samples were received from different geographical regions of the island. Of 377 samples tested, 94.2% were wild type (non-deletion mutant) homozygotes, 5.8% were delta 32 CCR5 heterozygotes, and none were delta 32 CCR5 homozygotes. The incidence of CCR5 delta 32/w heterozygous mutation among Puerto Ricans seems to be somewhat lower than what was reported with US Hispanics. Some age and gender associated bias of the mutation frequency were observed with the study population, the reason for which is unclear at present. PMID- 9734152 TI - RT-PCR comparative study of viral load levels in the HIV positive population in Puerto Rico before and after protease inhibitor regimen implanted. AB - Ponce School of Medicine AIDS Research Program conducted a large scale viral load assessment of Puerto Ricans who are infected by human immunodeficiency virus type 1 (HIV-1) during the summer of 1996 through the Roche ACCESS program before general implementation of combination therapy. Since January 1997, it has monitored those HIV-1 patients who are under treatments at most HIV-1 health care clinics, including both public and private. The present study was conducted to evaluate how the new treatment has generally impacted on the HIV-1 disease status of HIV-1 infected population in the eight Immunology Clinics. Assessment was made by consecutively monitoring the changes in HIV-1 viral load profiles of the population from January to September, 1997. A large majority of samples were delivered for viral load assessment without information of their treatment status, and only a small number of samples were identifiable either as baseline or followup. Despite the paucity of individual information, remarkable improvements of HIV-1 (+) population at large were evident. For example, in the summer of 1996 (ACCESS), population median viral load was 51,842; only 9% of the population had viral load less than 500 viral RNA copies/ml plasma and 72% had over 10,000 copies/ml. By July-September, 1997, the population median dropped to 8,679 (83%); 23% were below 500 copies/ml (+156%) and the proportion of patients who had over 10,000 copies/ml was reduced to 48% (-33%). The group of individuals who were positively identified as "follow-up" (i.e., under active treatment) had a median of 37128 copies/ml (-94%); 28% were below 500 copies/ml (+211%) and only 40% had more than 10,000 copies/ml (-44%). It is obvious that the implementation of triple combination therapy by PASET in 1997, has very markedly improve the HIV 1 disease status of HIV-1 (+) population in Puerto Rico. PMID- 9734153 TI - Anti-fungal and cytokine producing activities of CD8 + T lymphocytes from HIV-1 infected individuals. AB - Lymphokine activated killer (LAK) cells are capable of killing not only malignant cells but also hyphal form of Candida albicans in vitro. When peripheral blood mononuclear cells (PBMC) from normal healthy donors were cultured for 72-96 hrs with 1,500 international unit (IU)/ml interleukin-2 (IL-2), marked LAK activity was induced. However, even prior to IL-2 activation, PBMC isolated from some normal subjects and those from almost all individuals who are infected by human immunodeficiency virus type 1 (HIV-1) exhibited significant levels of anti-fungal activity. Such pre-activation ("in situ") antifungal activity of PBMC decreased during the initial 48 hrs of IL-2 activation. PBMC from HIV-1 seropositive subjects showed higher levels of "in situ" anti-fungal activity than normal PBMC did. After a decline of "in situ" activity during the initial 48 hours, LAK activity gradually increased and reached near maximal levels by day 4 and remained more or less constant until day 6. No significant difference was observed between the LAK activity of normal and HIV-1(+) PBMCs on days 4-6. In IL 2 activated normal and HIV-1(+) PBMC cultures, both CD4 and CD8 T cells produced IL-2, INF-gamma as well as TNF-alpha. Production of IL-2 by both CD4 and CD8 T cells was suppressed in HIV-1(+) PBMC cultures, but no significant suppression of INF-gamma production was noted. Meanwhile, TNF-alpha production by CD4 was very much suppressed but no significant changes in TNF-alpha production by CD8 T cells was noted in HIV-1(+) PBMC cultures. PMID- 9734154 TI - Fear research: implications for anxiety disorders. PMID- 9734155 TI - Caveolae a new subcellular transport organelle. AB - Recent advances have allowed the identification and characterization of well defined vesicular subcellular organelles involved in multiple basic cellular physiological processes, with demonstrated clinical relevance. Among these, three particular subcellular organelles have received special attention based on their proven and postulated participation in the sorting and targeting of small-and large-molecular weight molecules during exocytosis and endocytosis, and in cell signaling and transduction events. These have characteristic proteinaceous coat structures that allows their classification accordingly, into what has been described as clathrin coated vesicles and COP-coated vesicles and caveolae. In this review article a brief description of clathrin-coated vesicles and COP coated vesicles is presented. Caveolae (CAV), in turn, constitute a novel subcellular organelle that has received special attention based on its proven and postulated participation in transcytosis, potocytosis, and in cell signaling and transduction events. In this review of the literature a more extensive discussion is presented of CAV. In this context the article discusses the structural features of caveolae, its constituent protein caveolin(s), the functional aspects of this new organelle, and its postulated clinical relevance. PMID- 9734156 TI - Acute appendicitis: as a predisposing factor for acute glomerulonephritis--report of two cases. AB - This is a review of two children who developed acute glomerulonephritis (AGN) following acute gangrenous appendicitis (AGA) with periappendicular collections. The first patient presented with AGN during the course of appendicitis. The second patient developed AGN after appendectomy. Both patients did not have any other predisposing factors. AGN resolved in both patients after massive intravenous antibiotics. This is the first report of acute appendicitis as a predisposing factor for AGN. PMID- 9734157 TI - Unusual case of Meckel's diverticulum: a case report and review of an atypical form of presentation. AB - This is a review of a child who developed symptomatic anemia secondary to a huge Meckel's Diverticulum (MD). The patient presented with multiple complications, such as: neoplasia, occult chronic bleeding, giant size MD, partial intestinal obstruction and severe symptomatic anemia. There was complete resolution of the condition after resection and ileo-ileal anastomosis. After revision of the literature, this case is the first report of MD occurring concomitantly with such a myriad of signs and symptoms. PMID- 9734158 TI - What is your diagnosis. Effusive constrictive pericarditis. PMID- 9734159 TI - [Standardized patients in the evaluation of clinical competence]. PMID- 9734160 TI - The development of a course in basic physical examination skills. AB - Basic clinical skills of most medical school undergraduates continue unobserved and deficiencies have been detected in a significant number of physicians during residency. Nevertheless, our health care system is calling for competent graduates with solid basic clinical skills and a larger representation of qualified generalists in the increasingly important managed care environment. The need for a better introduction to Clinical Skills course was identified by students and clinical faculty at Ponce School of Medicine. In response to these concerns a new curriculum was developed with clear objectives, effective instructional strategies, and performance-based evaluation, with adult learning principles as its framework. The musculoskeletal examination unit of the curriculum was pilot tested and the course evaluation strategies revealed satisfaction with objectives, instructional and evaluation strategies, as well as improved confidence, and sense of usefulness for the learned skills. A curriculum in basic clinical skills that incorporates adult learning principles with solid instructional strategies can increase the confidence and skills of the learners and should lead to improved outcomes. PMID- 9734161 TI - [History of transplants in Puerto Rico]. PMID- 9734162 TI - [The first case of dissecting aneurysm of the descending aorta and bone marrow transplantation in Puerto Rico]. PMID- 9734163 TI - [Results of treatment of acute grade III acro-myo-clavicular dislocation by closed reduction and Kirschner wires fixation]. AB - Eight patients with acute, grade III acromioclavicular dislocation were treated by closed reduction and fixation with Kirschner wires introduced through the acromion into distal end of the clavicle. Desault plaster cast was discarded and K-wires removed after 6 weeks (in 3 patients after 4 weeks due to loosening). Minimum follow-up was 2 years. Patients were assessed with a 100 points scale of Kawabe et al., initial and final radiographs were compared. Four results were rated excellent (between 90 and 100 points), 3 results good (between 80 and 89 points) and in one case result was fair. Normal anatomy of acromioclavicular joint has been found in 75% of patients. Partial loss of reduction was found in two patients. It has been concluded, that closed reduction and fixation with Kirschner wires is simple, barely invasive method of treatment for acromioclavicular dislocation and gives good final results. PMID- 9734164 TI - [The use of radial artery for the revascularization of the amputated thumb]. AB - We present one of several methods for revascularization of the replanted thumb. Index radial artery is prepared, transposed and anastomosed to pollicis princeps artery. Twenty-seven patients aged 12-67 operated between 1991 and 1997 due to total or subtotal thumb amputation were included in this study. The method was used in 6 of them. In 21 patients different methods were applied to restore thumb arterial circulation. Venous grafts were used in 5 cases, end to end anastomosis of the artery in 16 cases. Thumb necrosis occurred in 2 cases in the venous grafts group and in 5 cases in end to end group. All thumbs revascularized with index radial artery healed successfully. The method requires only one microanastomosis and the risk of thrombosis is relatively small. The method should be recommended in cases of large arterial defects as an alternative for venous grafting. PMID- 9734165 TI - [Salvage surgery for severely mutilated hand: report of 2 cases]. AB - Immediate, one stage reconstruction is an optimal mode of treatment for severely mutilated hand. This might be feasible only with the use of tissues destined for amputation. Two cases are reported. The first patient had a thumb reconstruction with transfer of the index finger. The second one had metacarpal reconstruction with little finger structures. Both patients resumed previous occupation. PMID- 9734166 TI - [Hemispheric hydroxyapatite coated cups in total hip arthroplasty]. AB - The paper presents principles of implantation as well as clinical and radiological results of hemispheric hydroxyapatite coated cups use in total hip replacement. Eighty-seven patients (60 females, 27 males) aged from 16 to 72 years (mean 49 years) underwent 96 total hip replacements with the use of 51 ABG and 45 OCTOFIT hemispheric cups. Clinical results were satisfactory with an average Harris Hip Score of 89 to 91 at final follow-up. We have found total ingrowth and osteointegration of nearly all cups. In one case of acetabular reconstruction with bone grafts cup migration has been observed and revision was necessary. PMID- 9734167 TI - [The prospects of the usage of hip endoprosthesis in the fractures of femur in the elderly patients]. AB - Seven cases of comminuted, unstable trochanteric fracture of the femur treated with total or partial hip arthroplasty are presented. The age of patients at operation ranged from 69 to 88 years (mean 81.3). General condition of these patients was critical, the treatment used allowed for early mobilization. One patient died 9 weeks after surgery. In remaining 6 cases good functional result has been found at mean follow-up of 17 months (range 8 to 47 months). Hip joint arthroplasty for trochanteric fracture in elderly patients in poor general state allows for early mobilization. PMID- 9734168 TI - [Corticosteroid-induced non-vascular necrosis of the femoral head]. AB - The paper presents the issue of corticosteroid-induced avascular necrosis of the femoral head. Five patients (3 males and 2 females) 31 to 42 years of age (mean 34 years) are presented. Vascularized iliac bone graft was used in one case, cementless total hip replacement was done in the other. In remaining cases no surgery has been attempted. PMID- 9734169 TI - [The use of total cementless threaded knee Motta-Callea prosthesis]. AB - The paper presents features and operative technique for Motta-Callea knee arthroplasty. Medium-term results in a series of 17 joints with serious osteoarthritis (stage IV and V--classification of Larsen at al) in 14 patients aged 43-75 years (mean 64 years) are presented. Patients were evaluated according to Clinical Rating System (Insall et al.) before and after surgery. The average total score tripled after operation and was 168 of 200 points available. Good results have been achieved in both younger and elderly patients with advanced osteoarthritis and osteoporosis, also with bone plasty for varus or valgus deformity. An improvement was noted in regard of pain, alignment of the knee, its sagittal stability and walking ability, stairs included. PMID- 9734171 TI - [The evaluation of selected risk factors of solitary bone cyst recurrence after its curretage and bone graft filling]. AB - Selected risk factors of solitary bone cyst recurrence after curretage and bone graft filling have been analyzed in this study. Seventy-four patients (42 males, 32 females) have been reviewed. The age of patients at the diagnosis ranged between 4 and 55 years (mean 12 years). During follow-up ranging from 1 year to 48 years (mean 18 years) 15 recurrences appeared. It has been found, that sex of the patient, localization and volume of the cyst, septa presence or absence within it have no statistically significant influence on the recurrence. Age below 10 years and the use of allogenic lyophilized bone grafts predispose for recurrence of solitary cyst of bone. PMID- 9734170 TI - [The investigation of causes of treatment failures of osteosarcoma]. AB - A series of 57 patients (32 females, 25 males) aged 12-41 (mean 17.2) treated due to IIB advanced osteosarcoma has been analyzed. From among 20 patients who underwent surgery only, 2 persons survived. In 10 patients supplementary chemotherapy or preventive lung radiotherapy has been used. Two patients survived more than 5 years. In past 11 years 4 pre and postoperative chemotherapy protocols have been used. Big number of failures has been noted in the first few years. Despite chemotherapy recurrence took place in 6 cases. Failures resulted also from the late diagnosis or discontinued treatment. During last 5 years all 6 osteosarcoma patients received chemotherapy at Hematology and Pediatric Oncology Department and tumor resection has been performed. All patients stay alive (2 to 5 years). PMID- 9734172 TI - [The use of fluoride in the treatment of osteoporosis]. AB - Fluorides salts are used in the treatment for osteoporosis for 30 years, since their anabolic action on trabecular bone is well documented, but safety and efficacy of this treatment is still debated. Fluoride administration results in increased number and level of activity of osteoblasts leading to thicker trabeculae. The highest fluoride concentration and activity can be found within newly formed osteoid. Recommended dose is 15-25 mg per day, that is 33-55 mg of sodium fluoride or 140-190 of Na2FPo3. Calcium and vitamin D should be prescribed to all patients concurrently. Most frequent side effects include bone pain and dyspeptic problems. PMID- 9734173 TI - [Chronic bone abscess]. AB - The review of 68 patients with chronic bone abscess as a long-term sequel to ostomyelitis is presented. Antibiotic instillation within abscess cavity is recommended by the authors. Once the drainage is concluded immunotherapy should be introduced to secure long-lasting remission as in 90% of presented cases. PMID- 9734174 TI - [Patient-controlled analgesia (PCA) in postop pain management in scoliosis surgery]. AB - PCA method for postoperative pain management in scoliosis surgery is presented. If the child is capable of understanding the principle of its application and able to perform it the efficacy and safety of PCA depends on pump programming in regard of the single dose and time between doses. PCA has been used in 16 patients aged 11-16 years after C-D surgery for idiopathic scoliosis. Children with vital lung capacity below 50% of the normal value were excluded from this study. Single dose of morphine was between 0.015 to 0.03 mgkg body mass. Minimum time between the doses was 10-20 minutes. In all cases PCA resulted in satisfactory postoperative pain control. PMID- 9734175 TI - [Radial bone reconstruction with free vascularized fibular osteocutaneous graft: a case report]. AB - Radial shaft has been reconstructed in 14 years old boy with free vascularized graft. Fifteen by four cm skin island has been harvested along with the fibula and served to fill the defect within the forearm. Proximal end of radial artery has been microsurgically anastomosed to peroneal artery of the flap and peroneal vein branch with basilic vein. The bone healed after 3 months, remodeling took 5 months and overgrowth 8 months. The role of microsurgery in treatment for this type of injury has been emphasized. PMID- 9734176 TI - [Focal fibrocartilaginous dysplasia of the femur. A case report and review of the literature]. AB - A case of focal fibrocartilaginous dysplasia of the femur is presented. Varus femoral malalignment progressed from 30 degrees in 5th month of life to 51 degrees in 27th month of life. CT revealed flattening of the bone in the frontal plane, sclerotic marrow cavity with focal change resembling nidus of osteoma osteoides. Longitudinal defect filled with compact fibrous tissue was found below it. Corrective osteotomy has been done at the age of 32 months with good cosmetic result. PMID- 9734177 TI - [Report on 6-week visit of the Indiana Hand Center (Indianapolis, USA)]. PMID- 9734178 TI - Tuberculosis control in the Americas. PMID- 9734179 TI - El Nino and its impact on health. PMID- 9734180 TI - Eradication of foot-and-mouth disease in South America. PMID- 9734181 TI - [Functional surgery of the esophago-gastric junction: role of mini-invasive surgery]. PMID- 9734182 TI - [Immunohistochemical study on the prognostic value of the expression of laminin and Ki-67 receptors in advanced gastric cancer]. AB - The expression of 67-KDa laminin receptor (LR) was investigated in a group of 75 patients who underwent curative gastrectomy for advanced gastric cancer, with special reference to the possible role in the tumor progression and in the overall survival. In 56 out of these 75 patients also the prognostic significance of proliferative activity was investigated using the monoclonal antibody Ki-67. The tumor LR expression and the Ki-67 labeling index (Ki-67 LI) were immunohistochemically determined in paraffin-embedded sections using the avidin biotin immunoperoxidase method. The cumulative 5-years survival rate was 75.1% for patients without expression of LR, 52.6% for those with positive LR expression. Significant association between LR expression and depth of tumor invasion (p = 0.022) was found. By univariate analysis the presence of laminin receptor seemed to be associated with an higher risk of death (RR1.73-95% C.I. 0.71-4.20), but this effect disappeared after controlling for depth of tumor invasion. There was no significant relationship between the Ki-67 LI and wall invasion (p = 0.80) or nodal status (p = 0.73). The cumulative 5-year survival rates (95% CI) were 61.0% (35.3-79.2) in patients with Ki-67 index < 10%, 52.4% (29.7-70.9) with Ki-67 index = 10%-40%, 52.9% (27.6-73.0) with Ki-67 index > 40% and the differences were not statistically significant (p = 0.93). Also in multivariate analysis the proliferative activity did not independently affect survival (p = 0.98). An interaction between Ki-67 index and age was found and Ki 67 index > 40% was significantly associated with a poor prognosis in patients over 70 years old old (p = 0.002). In conclusion, tumor expression of laminin receptor could be correlated with gastric cancer aggressiveness, however its prognostic significance is already provided by depth of tumor invasion. The proliferative activity, determined with the monoclonal antibody Ki-67, does not seems to influence the survival except in elderly patients (> or = 70 years old). PMID- 9734183 TI - [Prospective research on fetal cholelithiasis: incidence, predisposing conditions, echographic diagnosis, and clinical features]. AB - The Authors report a prospective study on fetal cholelithiasis, analyzing its differences with the more widely known cholelithiasis of paediatric age. The study shows that the number of cases diagnosed by ultrasonography is higher than expected (0.39%, 3 cases on 764 pregnancies). The Authors could find no correlation between fetal cholelithiasis and any maternal, obstetrical and fetal factor. They have focused attention on sonographic imaging showing clinical instrumental correlation between echogenic material, clinical features and their evolution. The study confirms that the most common evolution results in spontaneous resolution of fetal endocholecystic pathological images. Finally, wide review of the international literature is reported on the rare, but possible clinical manifestations and their complications. PMID- 9734184 TI - [Hartmann's procedure in colorectal tumors. Evolutions of the indications in a series of 46 patients]. AB - The Authors report a retrospective study of 46 cases of Hartmann's operation in order to analyze the changing indications to this procedure in the management of colo-rectal cancer. The Hartmann's is operation has been performed in 46 out of 723 patients (6.4%) with colorectal cancer treated surgically from 1973 to 1997. Data concerning the indications have been analyzed in two consecutive periods, from 1973 to 1985 and from 1986 to 1997, respectively. In the first period, the procedure has been performed in patients with neoplastic perforation (40% of Hartmann's cases), and in an elective basis in patients with locally invasive tumor or intra-abdominal metastasis (20%). Indications for the procedure in the period 1986-1997 have been locally invasive tumor and/or distant metastasis (52.8% of Hartmann's procedures), neoplastic perforation (22.2%), high surgical anaesthesiologic risk (22.2%) or intestinal obstruction (2.8%). In the second period it has been noted a decrease of the number of patients that underwent Hartmann's procedure for bowel obstruction, and an increase in the number of cases in which the operation was performed for neoplastic perforation, for local and/or distant diffusion, or for high surgical risk. PMID- 9734185 TI - [Seminoma and adrenogenital syndrome]. AB - The Authors report a case of a 26 year old patient affected by adrenogenital syndrome, likely due to 21 hydroxylase defect, hermaphroditism (46XX genotype and female phenotype) and worked hyperandrogenism; moreover a hidden testis neoplasm (seminoma) was associated. PMID- 9734186 TI - [Incidentaloma of the adrenal glands: analysis of 9 surgical cases]. AB - In the assessment of incidentally discovered adrenal masses the detection of hormonal activity and the evaluation of benignity or malignity, either primary or metastatic, constitute the most important issues. This article reports 9 asymptomatic adrenal masses: The histopathological diagnosis consisted of cortical adenoma in 5 cases, adrenal metastatic mass in 2, respectively from a pulmonary microcytoma and from a renal carcinoma, a myelolipoma with leukemic infiltration and an hemorrhagic pseudocyst in the remnants. Evaluation of biochemical activity showed no endocrinological abnormality in all patients except in two cases of adenoma: the positivity of the 1 mg dexamethasone test, the low serum DHEAS levels and a concordant scintigraphic uptake were consistent with the pre-Cushing syndrome in the first case, whereas the ACTH inhibition revealed by low serum DHEAS levels without other hormonal alterations were the biochemical pattern in the second. Ultrasonography has been helpful in the diagnosis of adrenal mass in 6 cases, whereas CT scan allowed an etiopathogenetic diagnosis in 8 cases. All patients were submitted to adrenalectomy through the conventional surgical accesses; in 4 cases the adrenalectomy was performed as a associated intervention during vascular or gastrointestinal surgery. No postoperative death occurred. At follow-up ranging from 3 to 6 years, we recorded 4 deaths: the causes were represented by the progression of the primary malignancy in 2 patients that have been operated on for adrenal metastatic tumors, by hemorrhagic shock from an aorto-duodenal fistula and by systemic infectious complications respectively in the remnant two cases. The other patients were well and the endocrinological assessment showed normal findings. The Authors, according with data from literature, suggest an essential biochemical screening to evaluate the adrenal function in case of incidentally discovered mass: it is characterized by determination of plasma and urinary electrolytes, catecholamines, serum DHEAS and 17-OH progesterone levels, dexamethasone suppression test. In case of asymptomatic mass suspect for pheochromocytoma we advocate the MIBG scintigraphy.. The adrenocortical scintigraphy (NP 59) provides both anatomical and functional characterization of the adrenal glands: the concordant or discordant imaging patterns are useful in the diagnosis of benignity or malignancy. Although the management of patients with incidentally discovered masses remain controversial, we advocate adrenalectomy when they are hormonally hypersecreting, increasing in the diameter or malignant and in association with other abdominal operation. PMID- 9734187 TI - [Synchronous primary carcinoma of the colon. Diagnostic and therapeutic problems]. AB - Seven hundred and twenty-three patients with colorectal carcinoma were treated consecutively from November 1973 to April 1997. Seven patients (0.96%) were found to have two colorectal carcinomas (synchronous carcinoma), located in separated colonic areas. Clinical histories were analyzed with reference to sex, age, symptoms, physical findings, disease localization, pathologic classification, and survival data. Preoperative diagnosis of synchronous lesions is difficult, being achieved in only 2 cases, but it is important for the proper treatment of patients. It is concluded that full examination of the colon in all patients presenting with primary colorectal cancer is mandatory and that colonoscopy should be used to effectively screen patients for synchronous cancers. PMID- 9734188 TI - [Color Doppler echography in the study of pseudo-occlusion of the internal carotid]. AB - The differentiation of pseudo-occlusion from complete internal carotid artery occlusion may have important clinical consequences for patients with the former tend not to benefit from reconstructive surgery. The Authors report a case in which color-Doppler duplex-scanner revealed a persisting string-like lumen that was not demonstrated by angiography. The Authors believe that ultrasonography may in future permit the reliable differentiation of pseudo-from complete carotid occlusion, thereby reducing the need for angiography. PMID- 9734189 TI - [Treatment of post-phlebitic syndrome]. AB - The Authors report their experience in the treatment of the post phlebitic syndrome at ulceration stage. The therapy is based on three important points: a first medical treatment and later saphenous striping with a specific tying of the insufficient perforant veins to consolidate the results of scarification in the future; finally ther removal of plugs of fascia for the therapy of hypodermitis with a personal technique. PMID- 9734190 TI - [Carcinoma of the male breast: reconstructive technique]. AB - Male breast cancer is a rare neoplasm. Adhesions in its deeper and superficial levels and axillary adenopathies (50-60%) are often found at the diagnosis time. Surgery is considered the main step in the treatment of the male breast cancer. Radical mastectomy often leads to a wide asportation of the skin consequently causing some problems in the management of the chest-wall defect. Otherwise inadequate resections can cause local recurrences. The Authors, after a brief analysis of the main aspects concerning the prognosis and the management of the male breast cancer, report the use of transverse thoracoepigastric skin flap in the reconstruction of surgical wound after mastectomy. PMID- 9734191 TI - [Immunotoxicology in occupational and environmental medicine: prospectives, limitations, and research objectives]. AB - The immune system is able to recognize and neutralize potentially harmful agents, conferring to the organism resistance to infectious and malignant diseases. The authors have reviewed the literature and identified a group of substances able to enhance and/or reduce different immune functions, both in an experimental model and in occupational and environmental human exposure. The group includes several polyhalogenated hydrocarbons, particularly polychlorinated biphenyls, polybrominated biphenyls, tetrachloro-dibenzo-p-dioxin (TCDD), some metals like lead, cadmium and mercury, pesticides, i.e. dithiocarbamates and organotin compounds, organic solvents. The observed changes are usually slight and do not allow prognostic conclusion. In this study, the authors propose a 3-level rank of tests suitable for the immune evaluation of individuals occupationally exposed to xenobiotics, divided into three levels, as follows: tier 1: immunoglobulin classes (IgG, IgA, IgM), complement fractions (C3, C4), rheumatoid factor, and non-organ specific autoantibodies (AMA, SMA, ANA); CD3, CD4, CD8, CD57, CD20, HLA DR; CD3/HLA-DR positive lymphocyte subsets; tier 2: determination of the mitotic response of peripheral blood lymphocytes to phytohaemoagglutinin, anti-CD3 monoclonal antibody, phorbol-myristate-acetate (PMA) and polyclonal immunoglobulin production after stimulation with pokeweed mitogen; tier 3: cytokine production with and without mitogen stimulation. The approach is "step by step" and assumes the need of a closely integrated and comparative evaluation of the findings obtained. The protocol could be used in research fields; moreover, some of the tests could be useful in the monitoring of persons exposed in the environment or in the workplace to immunotoxic substances or to biological agents. PMID- 9734192 TI - [Asthma and rhinoconjunctivitis caused by rape flour: description of a clinical case]. AB - A 48 year old man, employed in a grain and animal feed store for 9 years, was referred to our clinic complaining of nasal blockage, rhinorrhea, sneezing, ocular burning, coughing and wheezing occurring over the last 12 months. The man's main task was to manually load and unload the unpackaged grain and feed. Symptoms occurred only when he worked directly with oilseed rape flour and not when he worked with other types of grains. Eye and nasal symptoms appeared during the work shift, while respiratory symptoms were worse at night than during the day after exposure to rape for more than 2 consecutive days. Physical examination was normal, as were the results of the pulmonary function studies. The methacholine inhalation test, performed to measure the level of non-specific airways responsiveness, showed normal bronchial reactivity. Results of allergy skin prick tests were negative for common inhalant and food allergens, but slightly positive for the oilseed rape flour extract. Registration of the peak expiratory flow (PEF) showed slight decreases in PEF values after occupational exposure. We conclude that this case is suggestive of asthma and rhino conjunctivitis, induced by oilseed rape flour, probably due to an allergic mechanism. PMID- 9734193 TI - [Dental amalgams and urine elimination of mercury in workers exposed to low concentrations of inorganic mercury]. AB - The aim of the research was to assess the contribution of dental amalgams and other non-occupational factors of exposure to inorganic mercury (diet, etc.) to the quantity of mercury excreted with urine in workers exposed to low level concentrations of inorganic mercury. Two groups of workers (Groups I and II) were studied who were exposed to low and different environmental concentrations of inorganic mercury. These two groups were compared with a group of subjects not occupationally exposed to mercury in the same geographical area (Group III). All subjects were administered a questionnaire concerning personal data, lifestyle, recent removal and/or insertion of dental amalgam fillings, presence of nasal obstruction or bruxism and consumption of fish. The number of amalgam-filled teeth was established for each subject. Mean environmental exposure to inorganic mercury was 0.0087 mg/m3 for Group I and 0.0030 mg/m3 for Group II. Urinary excretion in the 3 groups was 4.2 +/- 2.8 micrograms/l for Group I, 3.0 +/- 2.1 micrograms/l for Group II and 1.6 +/- 1.2 micrograms/l for Group III. The results showed that of the factors of exposure to inorganic mercury, only occupational exposure (T = 9.18; p = 0.000) and the number of amalgam-filled teeth (T = 2.03; p = 0.043) were able to influence significantly urinary excretion of mercury; the sources of non-occupational exposure did not appear to play any role. The contribution of each amalgam filling to urinary mercury excretion was calculated to be 0.08 microgram/l. Occupational exposure therefore, even at low level doses, is still the main cause of urinary mercury excretion in workers exposed to inorganic mercury; of the non-occupational exposure factors, a significant role is played by amalgam dental fillings, whose contribution needs to be taken into consideration in order to make a correct interpretation of the results of biological monitoring of exposed workers. PMID- 9734194 TI - [Frequency of lumbago in a cohort of nursing students]. AB - The etiology and frequency of low back pain among health care personnel have been widely studied by means of cross sectional studies. The aim of our study was to calculate low back pain incidence in a prospective cohort of nursing students. A population of 344 subjects (72 males and 272 females) was involved in this investigation. Every student was submitted to a clinical and functional examination of the spine before beginning training and was checked in two following steps by a specific questionnaire for epidemiological studies of spinal disorders in working communities. 197 subjects (57.3%)(41 males and 156 females) completed follow-up. The low back pain incidence was similar at the end of two exposure periods (12.1% and 13.1%). The cumulative incidence was 22.5% throughout the study period. The longitudinal study allowed good control of selection bias and confounding factors; more over it showed that compared to other measurements of occurrence the cumulative incidence was between other occurrence measures, more informative, in our case, than prevalence and incidence rates. A cumulative incidence of low back pain over 20% after only two years of exposure in a young and healthy population of nursing students, requires implementation of ergonomic measures for patient handling tasks. PMID- 9734195 TI - An alternative method of detecting dispersion staining colors for the determination of asbestos fibers in bulk materials. AB - Asbestiform mineral particles can be detected and identified by means of a polarized light microscope, making use of the dispersion staining technique. In order to obtain dispersion staining (central stop), che rays which pass undeviated through liquid and specimen must be stopped. A microscope objective can be fitted with a device which can insert a central stop on the objective back focal plane to prevent undeviated rays from reaching the objective. Alternatively, the sample can be observed in a conventional dark field set-up, that is, it can be illuminated by a hollow cone of light of greater aperture than the microscope objective so that only rays which are strongly deviated can enter the objective. The first method, proposed by McCrone and widely used in the USA, allows very bright and defined colors to be obtained but at the same time entails low resolution. The second method, recommended by italian regulations as a method that can be used in asbestos characterization, produces less defined and fainter colors. By suitably modifying the angular aperture of the illuminating cone and regulating the distance between condenser and specimen, good results were achieved: on the one hand a good image resolution were retained, on the other, the colors observed were bright and well defined. The detection of these colors can represent a fundamental step in the characterization of the specimen under analysis. PMID- 9734196 TI - Interconversion of units of three methods for blood cholinesterases. PMID- 9734197 TI - [If I were to wake-up clumsy and trembling...(or how to begin treatment of Parkinson disease]. PMID- 9734198 TI - [Digit span, automatic speech and orientation: amplified norms of the Barcelona Test]. AB - BACKGROUND: Digit span, automatic speech and orientation are screening tests of interest in neurology due to their high sensitivity in cerebral pathologies and easy applicability. These tasks assess attention, concentration, mental control, orientation and short-term memory. OBJECTIVES: To analyze the impact of age and education on the results of the following subtest included in the Barcelona Test: digit span, automatic speech and orientation. PATIENTS AND METHODS: 275 normal subjects were evaluated. Mean (SD) age: 52 (17.7), range (18-19). Mean education: 9 (5.5) years, range (0-25). Subjects were stratified into 7 groups by age and education. Nine items of Barcelona Test were used: digits span forward and backward, automatic speech, forward and backward, with and without time, and person, time and place orientation. RESULTS: Statistical description, parametric and non parametric test. The Kruskal Wallis test was used, showing that aged and education are significant difference in the automatic speech forward or backward, with or without time, while it did not showed a significant difference in the different groups in the orientation tests. Analysis of Variance for group of age and education showed significant differences in the digit span forward and backward (F = 19.92; p < 0.001; F = 18.44, p < 0.001), respectively. CONCLUSIONS: Both age (inversely) and education (directly) influenced on the scores of the analyzed items, except to person, place and time orientation that are not affected for them. PMID- 9734199 TI - [Abstract abilities: amplified norms for the Barcelona Test]. AB - INTRODUCTION: By abstract abilities is understood the ability of comprehend relations identifying the essential components and taking out from them a common feature. Its evaluation constitutes a great challenge for neuropsychology. Several studies have showed the influence of age and formal education in the performance of tests assessing these abilities. AIM: To analyse the WAIS-style abstract abilities included in the abbreviated version of the Barcelona Test determining the influence of and formal education and obtaining more normative data for a Spanish population. METHOD: A sample of 264 subjects was randomly selected. The subtests of the Barcelona Test were administered in a single session to every subject. We selected the scores obtained in Arithmetic Problems (raw and timed), Similarities, Digit Symbol and Block Design (raw and timed). RESULTS: The analysis of the variance showed statistically significant results of the selected items according to age and formal education. The formal education, as the lineal regression showed, resulted significant for all the items, being the Arithmetic Problems the only subtests not influenced significantly by the formal education. CONCLUSION: The results obtained in this study are in general concordant to the ones of the reviewed bibliography. PMID- 9734200 TI - [Spanish study of quality of life in migraine (I). Profile of the patient with migraine attending neurology clinics]. AB - BACKGROUND: Migraine is the main reason for neurological consultation. OBJECTIVES: To analyse the profile of the patient with migraine attending the Neurological Services of our country. PATIENTS AND METHODS: Neurologists from 7 hospitals in different spanish regions interviewed 305 patients (at least 40 per hospital) who met migraine diagnostic criteria. They used an ad hoc questionnaire in which detailed demographic and migraine clinical data were included. Patients with transformed migraine or tension-type headache more than two days per week were excluded. RESULTS: The majority (82%), were women, with no other diseases, with an average social (88%) and cultural (41%) level. The mean age at consultation was 38 +/- 11 years, while the mean duration of migraine history was 18 +/- 13 years; 78% met criteria of migraine without aura, 15% of migraine with aura and the remaining (8%) both migraine with and without aura criteria. Main subjective precipitating factors were: stress (80%), foods (68%), drugs (34%), alcohol (20%) and menstruation (8%). Migraine pain was referred to as mild by 4% of cases, as moderate by 59% and as severe in the remaining 37%. The usual duration of migraine attacks ranged from 12 to 24 h in 35% of cases, from 24 to 48 h in 25%, from 4 to 12 h in 23% and was longer than 48 h in the remaining 17%. More than half (53%) had more than 3 attacks per month. The pain was unilateral in 70% of cases, and more than half had vomiting (57%) and sono and/or photophobia (97%). CONCLUSIONS: The typical profile of the migraine patient attending the Neurology Services in Spain is that of a woman aged from 20-50, with a long personal history of migraine, otherwise healthy and with an average socioeconomic and cultural level. Our data confirm that migraine attacks are incapacitating in a relevant number of these patients. PMID- 9734202 TI - [Giant cavernous angioma]. PMID- 9734201 TI - [Vitamin E and Parkinson disease]. PMID- 9734203 TI - [Obstructive hydrocephalus in a patient with Paget's disease]. PMID- 9734204 TI - [Spontaneous dissection of extra-intracranial intern carotid]. AB - Dissection of internal carotid artery is an unusual cause of stroke. It generally affects the extracranial portion of the vessel, rarely the intra-cranial portion and exceptionally both sections simultaneously. We present two cases of spontaneous dissection with extra and intra-cranial involvement. Two females, 46 and 36 years old, presented as stroke of the right internal carotid (ICA) associated with headaches and ipsilateral Horner's syndrome. An echo-Doppler was done on the first patient, which turned to be normal, and carotid angiography was done to both patients. The first patient showed a filiform stenosis of the right ICA that ran from the origin to the carotid siphon. The second patient showed a longitudinal stenosis of the right ICA 2 cm from the origin, which ended in an obstruction of the terminal branches. The control angiographs at five and six months respectively, showed partial re-channelling or complete re-channelling. The first case was treated with anti-aggregants and the second with anticoagulants. There were no new episodes in either cases. Dissection of the ICA usually only affects the extracranial portion of the artery, stopping in the petrous portion. We do not know why dissection also affected the intra-cranial section of the artery in these two cases. PMID- 9734205 TI - [Post-radiotherapy brachial plexopathy and cervical myelopathy]. AB - The case of a 58-year-old male who, after 47 months of treatment with cervical radiotherapy for labial carcinoma, developed brachial plexopathy and six months later cervical myelopathy is herewith presented. Involvement of both the brachial plexus and the spinal cord in the same patient secondary to radiotherapy is exceptional. PMID- 9734206 TI - [Anesthesia paresthetica: contribution of a new case and evolutive study using magnetic resonance]. AB - Patients with subclinical cobalamin deficiency may develop a subacute myeloneuropathy after nitrous oxide anesthesia. Frequently they have hematologic manifestations, however there is only a few reports of spinal lesions shown on MR and particularly of the effects of cobalamin replacement therapy on these lesions. We describe a patient with unsuspected cobalamin deficiency who developed subacute combined degeneration of the spinal cord after nitrous oxide anesthesia. MR showed high intensity in dorsal columns on T2-weighted image, and posterior resolution of abnormalities after therapy. We discuss the importance of preoperative hematologic manifestations for select high risk patients and prevent this complication. On the other hand, we consider MR as a useful tool to improve our knowledge in the pathophysiology of the neurologic manifestations of the process. PMID- 9734207 TI - [Unilateral lumbosacral plexus neuritis after herpes zoster infection. Favorable response to prednisone]. PMID- 9734208 TI - [Respiratory failure secondary to dysautonomy in lateral bulbar infarction]. PMID- 9734209 TI - [Abdominal tumor presenting as segmental weakness caused by zoster]. PMID- 9734210 TI - [Brain stem auditory evoked potentials in meningitis]. PMID- 9734211 TI - [Bacterial endotoxins and their effects]. AB - The lipopolysaccharide endotoxin macromolecules are cell wall's components of the Gram negative bacteria. The endotoxins are produced by Gram negative bacteria of intestinal flora. If the endotoxins are translocated from the intestinal tract to the circulation or injected into bloodstream, they elicit (depending from the quantity of endotoxin), slight or serious effects (e.g. endotoxin shock). In the effects of endotoxin certain cell populations (e.g. thrombocytes, macrophages, leukocytes, etc.), certain organs and organ-systems (e.g. liver, spleen, bone marrow, endocrine and lymphoreticular systems etc.) are involved. Effects of endotoxin are produced by mediators (e.g. endotoxin binding proteins, cytokines, prostaglandins, prostacyclins, NO etc.). The endotoxin sensitivity of vertebrate organisms is dependent from the phylogenetical status of the species. Most sensitive species is the human. Generally accepted that endotoxin has an important role in the pathogenesis of septic shock. In other pathological processes (e.g. intestinal syndrome of radiation disease, Gram negative infections, various shock forms etc.) are supposed or proved the role of endotoxins. Lead acetate induced endotoxin hypersensitivity or LAL methods are good tools for demonstration of the role of endotoxin in the pathogenesis of various processes. For this reason, the experimental endotoxin shock is used a model of septic and other shocks. PMID- 9734212 TI - [Histological characteristics of chronic hepatitis C in biopsy material]. AB - BACKGROUND/AIMS: Hepatitis C virus (HCV) infection is one of the most important diseases with high chronicity rate (50-80%) leading to end-stag cirrhosis and hepatocellular carcinoma. Hepatic histology shows a characteristic but not diagnostic picture. The aim of this study was to evaluate the characteristic histological findings in correlation with epidemiological features in our liver biopsy material. PATIENTS/METHODS: 106 liver biopsies were studid between 1993 1996. All patients (60 males, 46 females, age between 11-81 years, mean age: 43 years) were found to be positive for HCV antibody by a second-generation ELISA method. The biopsy materials were fixed in buffered formalin and having embedded in paraffin, stained with hematoxylin and eosin, periodic acid-Schiff after diastase digestion, Gomori's reticulin stain and picrosirius red for collagen. The histological evaluation was based upon the new classification of chronic hepatitis proposed by Desmet et al. The statistical analysis was performed by the Chi square test. RESULTS: Minimal chronic hepatitis (HAI: 1-3) was found in 14 (13.2%), mild chronic hepatitis (HAI: 4-8) in 69 (65.09%) and moderate chronic hepatitis (HAI: 9-12) in 23 (21.69%) cases, while assessment of fibrosis (staging) resulted fibrosis 0/1 in 44 (41.5%), fibrosis 2 in 14 (13.2%), fibrosis 3 in 37 (34.9%) and cirrhosis (fibrosis 4) in 11 (10.37%) cases. Among histological features of chronic hepatitis C, the frequency of steatosis (70.75%), lymphoid F/A (63.2%), and bile duct lesions (12.26%) have paralelly increased with activity (grade) of hepatitis and these changes were more pronounced in moderate chronic hepatitis (p < 0.001). CONCLUSIONS: More than half of chronic hepatitis C patients presented mild histological lesions with stage 1 fibrosis. Lymphoid F/A, bile duct damage and steatosis are important diagnostic features that show a strong correlation with the activity of chronic hepatitis. The assessment of fibrosis (stage: 3 and stage: 4) in mild chronic hepatitis cases does alert the hepatologist to perform the liver biopsy to detect the fibrotic changes in chronic hepatitis C. PMID- 9734213 TI - [The O'Brien filter test in the differential diagnosis of disorders with high grade thrombocytosis]. AB - In the differential diagnosis of primary and secondary thrombocytosis, platelet function test can be used. We have examined the possible role of O'Brien's filter test in the differentiation of primary and secondary thrombocytosis in 53 patients with myeloproliferative diseases with primary thrombocytosis and in 21 patients with other disorders complicated by secondary thrombocytosis. By using heparin as an anticoagulant, the sensitivity of O'Brien's filter test proved to be 75%, and it's specificity was 85.7%. In blood samples anticoagulated with citrate, the sensitivity was 100% and specificity 83.3%. Based on these studies we suggest the use of O'Brien's filterometer as a screening test in the differential diagnosis in patients with elevated (> 400 x 10(9)/L) platelet count. In case of normal results, the causes of reactive thrombocytosis should be cleared first, while with pathologic results, haematological examination of the patients should be performed. PMID- 9734214 TI - [Clinical and molecular genetics of hypertrophic cardiomyopathy]. AB - Recent developments in molecular genetics have allowed to identify mutations in seven genes coding the beta myosin heavy chain, troponin T, alpha tropomyosin, myosin binding protein C, essential and regulatory myosin light chains and troponin I causing hypertrophic cardiomyopathy. These mutations affect critical, evolutionary conserved nucleotides of these genes and influence vital functions of the encoded proteins. As all seven genes encodes sarcomeric proteins in the heart muscle, hypertrophic cardiomyopathy is regarded these days as a disease of the sarcomer. Recent data indicate that some mutations are associated with "malignant" clinical picture, with rapidly developing, severe symptoms of the disease and increased risk of sudden cardiac death while other mutations bear a more favourable prognosis. Apart of the disease causing mutation other factors, including disease modifier genes, are likely to make an impact on the clinical appearance of hypertrophic cardiomyopathy. The knowledge provided by molecular genetics influences the clinical management of the disease even today and based on the investigation of mutation carrying patients new diagnostic criteria was proposed for hypertrophic cardiomyopathy. The challenge for the future is the establishment of routine genetic diagnostics and the development of possible gene therapy. PMID- 9734215 TI - [Nosocomial ornithosis]. AB - Ornithosis is an occupational hazard to workers in the poultry industry, zoonosis. Own sporadic unusual case is appearing under the hospital circumstances as a nosocomial atypical pneumonia. PMID- 9734216 TI - [Viral contamination of food products: a poorly understood public health problem]. AB - Throughout the world there have been several epidemics of food-borne diseases (FBD) about which there is lack of sufficient information for public health institutions to take appropriate measures. This study was conducted for the purpose of contributing to the dissemination of information on these diseases and their etiologic agents, epidemiology, and control. The study was based on data from 61 sources, including review articles, reports of outbreaks, and databases. Results reveal considerable underregistration and lack of data on FBD throughout the various countries, with viruses being the second most important cause of FBD in the United States of America. Two agents, Norwalk virus and hepatitis A virus, were the fifth and sixth most frequent causes, respectively, although the former was the single most frequent cause of FBD in 1982 and the second most frequent cause of water-borne diseases during the period from 1986 to 1988. Despite the scarcity of information on the problem, rotavirus, poliovirus, hepatitis E virus, astrovirus, and small gastroenteric viruses are also important causes of FBD. We also discuss the importance of viral zoonoses, especially hemorrhagic fevers transmitted by contact with rodent feces and tick-borne viral encephalitides (Lassa fever). There is discussion of the controversial mad cow disease and its potential transmission through food products, as well as of dietary aspects of the management of AIDS and other viral infections. Finally, measures for the prevention and control of FBD are described. PMID- 9734218 TI - [Non-prescription drugs in Brazil]. AB - In this study we examined the 100 over-the-counter drugs that were most widely sold in Brazil from June 1992 to June 1993. We excluded 23 food products that were classified as medicinal. The sample, which included 77 drugs, was representative of about 67% of all sales in monetary value, and 76.8% of all units sold of the different classes of over-the-counter drugs. The anatomical therapeutic-chemical (ATC) system was used to classify the various drugs; each class of drugs was also graded according to a scale of its intrinsic value, taking into account effectiveness and risks. Most classes of drugs (91%) showed "little intrinsic value" (that is, were of questionable or no value, of relative value, or unacceptable), and 70% were fixed-dose combinations. Only 10 classes of drugs were included in the Ministry of Health's National Drug List, and four appeared in WHO's list of essential drugs. The therapeutic classes to which the drugs belonged were, in descending order of frequency, for the digestive tract, skin, genitourinary system, musculo-skeletal system, central nervous system, parasitic diseases, respiratory tract, and sensory system. The therapeutic subgroup that commanded the highest sales was that of the non-opiate analgesics and antipyretics. Our results confirm the hypothesis that over-the-counter drugs in Brazil are of poor therapeutic quality and that the use of many such drugs is a source of unnecessary expense for the population. Some of them should not be accessible to individuals who lack current knowledge of their side effects, since 25% of all cases of poisoning in the country are related to self-medication. Modifying the public's perception of the need for taking drugs to relieve their ailments is one approach that may improve their use of medications in the long term. PMID- 9734217 TI - The Onchocerciasis Elimination Program for the Americas: a history of partnership. AB - The decision in 1987 by the pharmaceutical firm Merck & Co. to provide Mectizan (ivermectin) free of charge to river blindness control programs has challenged the international public health community to find effective ways to distribute the drug to rural populations most affected by onchocerciasis. In the Americas, PAHO responded to that challenge by calling for the elimination of all morbidity from onchocerciasis from the Region by the year 2007 through mass distribution of ivermectin. Since 1991, a multinational, multiagency partnership (consisting of PAHO, the endemic countries, nongovernmental development organizations, the Centers for Disease Control and Prevention in Atlanta, Georgia, as well as academic institutions and funding agencies) has developed the political, financial, and technical support needed to move toward the realization of that goal. This partnership is embodied in the Onchocerciasis Elimination Program for the Americas (OEPA), which is supported by the River Blindness Foundation (RBF) and now by the Carter Center. OEPA was conceived as a means of maintaining a regional initiative to eliminate what is otherwise a low priority disease. Since its inception in 1993, the OEPA has provided more than US$ 2 million in financial, managerial, and technical assistance to stimulate and/or support programs in Brazil, Colombia, Ecuador, Guatemala, Mexico, and Venezuela, so as to take full advantage of the Merck donation. Now halfway into a five-year, US$ 4 million grant provided through the Inter-American Development Bank, the OEPA's capacity to support the regional initiative is assured through 1999. PMID- 9734219 TI - Childhood asthma along the United States/Mexico border: hospitalizations and air quality in two California counties. AB - Since the passage of the North American Free Trade Agreement in 1993, there has been an increasing need to monitor environmental health trends that may be related to the rapid industrialization of the United States/Mexico border. We studied two counties on the California/Baja California border to obtain baseline data on trends in childhood asthma hospitalizations and two pollutants that aggravate asthma, ozone and particulate matter (less than 10 microns in diameter), from 1983 to 1994. Hospital discharge records of children 14 years and younger were analyzed, and rates by county, race, and sex were age-adjusted to the 1990 California population. Data on five ozone and particulate matter indices obtained from the California Environmental Protection Agency were used. Imperial County had the highest childhood asthma hospitalization rates in California for non-Hispanic whites and African-Americans, and the second highest for Hispanics. San Diego County had rates below the state average. Over the time period examined, rates in Imperial County increased 59%, while those in San Diego County decreased 9%. Maximum ozone levels increased 64% in Imperial County but decreased 46% in San Diego County. Particulate matter levels were four times higher in Imperial than in San Diego County. High rates of childhood asthma hospitalizations in Imperial County may be partially related to high levels of poverty and worsening air quality conditions produced by increased burdens on the local airshed. Asthma prevalence surveys and binational time-series analyses examining asthma-pollutant relationships are needed. PMID- 9734220 TI - Medical genetic services in Latin America: report of a meeting of experts. PMID- 9734221 TI - [Polymerase chain reaction for rapid detection and serotyping of dengue virus in clinical samples]. AB - This study describes the benefits of using reverse transcriptase polymerase chain reaction (RT-PCR) for the rapid detection and typing of dengue virus in clinical samples. Twenty-seven serum specimens from patients with dengue fever and dengue hemorrhagic fever in Colombia, Nicaragua, and Panama were directly subjected to RT-PCR for the detection of dengue virus. The resulting double-stranded DNA product was typed by a second round of PCR amplification (nested PCR) with type specific primers, viral culture/indirect immunofluorescence (IIF), and enzyme linked electroimmunoassay for IgM anti-dengue antibodies. The amplified virus genome was detected and typed within 8 hours. Nested RT-PCR, using viral culture and IIF as the gold standard, showed 100% sensitivity; 78% specificity; 69% positive predictive value, and 100% negative predictive value. It is noteworthy that two of the specimens whose results were positive with nested RT-PCR and negative with viral culture showed specific IgM antibodies. The results of the RT PCR were in close agreement with those obtained through viral culture. This suggests PCR can greatly facilitate the rapid and early diagnosis of dengue infection. PMID- 9734222 TI - [Early diagnosis of Hansen disease: study of the health services in Recife (Pernambuco), Brazil]. AB - This paper presents the results of a descriptive study carried out in the city of Recife, state of Pernambuco, Brazil, between March and September 1994. The study aimed at health services available for performing early diagnosis of Hansen's disease with emphasis on accessibility and quality of the services provided. The sample consisted of 32 health clinics visited for diagnostic purposes by 183 patients with Hansen's disease. Information on organizational infrastructures was collected by means of interviews with health clinic managers. Information regarding routine procedures in the 32 clinics was collected by observation, with special attention given to archival and inspection activities. A total of 1,998 patients were interviewed to determine accessibility of services. Time spent in consultation with the physician was determined for 1,000 patients who were seen by 123 physicians at the clinics during the interviews. To explore physicians' attitude and knowledge regarding Hansen's disease, 133 were randomly selected from a list of names. The following factors were identified as hindering early diagnosis of Hansen's disease: the large number of people seeking service who could not be seen by a physician on the same day; the long time elapsed between appointment scheduling and the actual visit (for those not seen on the same day); the long wait for the consultation; the brevity of the consultation; the low availability of trained personnel; the low proportion of physicians who examined all body surfaces; difficulties in the clinical recognition of the disease; and physicians not prepared to make a differential diagnosis. These obstacles can precipitate the physical deterioration of Hansen's disease patients and stimulate the persistence of transmissibility; therefore, they need to be overcome if Hansen's disease is to be eliminated. PMID- 9734223 TI - HIV/AIDS practice patterns, knowledge, and educational needs among Hispanic clinicians in Texas, USA, and Nuevo Leon, Mexico. AB - Hispanic clinicians in Texas, United States of America, and in the neighboring state of Nuevo Leon, Mexico, were surveyed to determine their educational needs in the area of HIV/AIDS. Two-thirds of the 74 Texan and 22% of the 104 Mexican physicians queried had seen at least one HIV/AIDS patient in the previous year. The majority of the respondents were primary care physicians who: 1) were in private practice; 2) saw more than 1,000 patients per year; 3) had been out of training for more than 10 years; 4) provided some HIV prevention education to patients based on their perceived risk of infection; 5) rated their own knowledge of HIV/AIDS as average but rated their knowledge of treatments for the disease below average; 6) received most of their information about HIV/AIDS from journals rather than formal continuing education programs; 7) thought Hispanic patients had special needs with regard to HIV/AIDS care; and 8) were willing to attend education programs to improve their HIV/AIDS management skills. The greatest barriers to caring for HIV patients were lack of clinical knowledge and fear of infection. These results point to a need for a large-scale training program to improve the HIV/AIDS management skills of Hispanic clinicians in Texas and Nuevo Leon. PMID- 9734224 TI - A prefilled injection device for outreach tetanus immunization by Bolivian traditional birth attendants. AB - This study evaluated the performance, acceptability, and appropriateness of a new, single-use, prefilled injection device called UniJect for an outreach immunization application. Between April and June 1995, UniJect devices were used by 36 traditional birth attendants to administer tetanus toxoid injections to 2,240 pregnant women during routine, antenatal home visits in the Northern, Ichilos, and Warnes Districts of Santa Cruz, Bolivia. Because tetanus toxoid is relatively heat stable, the traditional birth attendants were able to keep the tetanus toxoid-filled UniJect devices in their homes for up to one month without refrigeration. The devices were stored, transported, and disposed of in an outreach carrier designed to reduce the risks of improper handling and disposal. Data were collected from injection recipients, traditional birth attendants, and supervisors via observation, questionnaires, and post-study interviews. The performance of the UniJect device and its acceptability among all groups was very high. The traditional birth attendants used UniJect properly and safely; there were no reports or observations of device misuse, reuse, or needle-stick. Advantages cited included the fact that the device required no assembly, offered assured sterility, and reduced vaccine wastage sometimes associated with multi dose vials. The ability to store and transport the vaccine-filled devices without ice also greatly simplified logistics. PMID- 9734225 TI - [Indications for episiotomy at public maternity clinics in Nequen, Argentina]. AB - Indications for performing episiotomy during vaginal births are a controversial topic requiring worldwide review. In Neuquen Province, Argentina, when standards for low-risk childbirth were developed in 1995, they included the provision to limit episiotomies to cases at high risk for spontaneous laceration. The present retrospective cohort study was designed for comparing the criteria applied in 1996 as indications for episiotomy in public maternity services of Neuquen Province, as well as the effect of parity and other variables on its frequency. The Perinatal Data System yielded 7,513 medical records for review, which represented 70% of all the institutional births during the year. Of these records, 830 dealing with cesarean sections did not qualify for the study. The remaining 6,683 records were divided into eight groups according to hospital location. Episiotomy incidence rates were estimated for those eight groups and the Poisson regression was applied in order to adjust for birthweight, number of siblings, mother's age, and type of birth presentation and outcome. Taking the Neuquen Hospital data as baseline because of its higher complexity and large number of births taking place there, two hospitals had episiotomy incidence rates equal to 70% (95% CI: 62%-79%) and 67% (95% CI: 57%-78%) of the Neuquen Hospital rates. Two other hospitals had incidence rates which were higher by 28% (95% CI: 13%-45%) and 17% (95% CI: 2-35%), while the remaining hospitals showed no significant differences. Stratified regression by number of previous vaginal births showed parity to be the strongest influencing variable on indications for episiotomy. Rates for nulliparous women showed no differences by hospital, but rates for primiparous women did, with even stronger differences shown for multiparous women. The authors concluded that all institutions included in the study performed episiotomies as a virtually routine procedure on nulliparous women, that there are significant differences in their indications for primiparous women, and that those differences increase along with parity. These differences seem to show that maternity clinics in the Province of Neuquen vary widely in their criteria for episiotomy indications. PMID- 9734226 TI - [Evaluation of a supplementary nutrition program]. AB - The objective of this study was to evaluate the impact of a feeding supplementation program on the growth of undernourished children younger than 5 years in the city of Guariba, state of Sao Paulo, Brazil. The sample consisted of 469 malnourished children enrolled in a feeding supplementation program sponsored by the State Health Secretariat. The children were divided into four groups according to how long they had been enrolled in the program: in group 1, the children had been enrolled for up to 12 months; in group 2, from 12 to 24 months; in group 3 from 24 to 36 months; and in group 4 the children had been enrolled for more than 35 months. Percentiles for weight/age, height/age and weight/height were calculated for each child. To assess the impact of the program, reference curves for the anthropometric profile were constructed based on expected variations in population percentiles. The changes observed in each group were analyzed statistically (McNemar). Groups 1 and 2 presented weight recovery and gains in the weight/height ratios for the most severely malnourished children; in group 3, the weight/height ratio was maintained and there was a discrete tendency towards weight recovery, which was reversed in group 4, in which the weight was again low in relation to height. The feeding supplementation program temporarily minimized severe nutritional deficiencies but was not sufficient to recover and maintain normal growth. PMID- 9734227 TI - Increased prevalence of Plasmodium falciparum malaria in Honduras, Central America. AB - We report on our investigation of a malaria outbreak in Honduras, Central America, in January 1997. We tested 202 patients with fever and chills using thin and thick blood film microscopy. Sixteen patients lived in the city and the rest lived in rural areas. A total of 95 samples (47%) were positive for malaria parasites. Seventy-nine percent (63/80) of the rural patients were infected with Plasmodium vivax and 21% (17/80) were infected with P. falciparum. In the urban area, all 15 infected patients had P. vivax malaria and none showed evidence of P. falciparum. Since previous reports indicate that falciparum malaria accounts for only 2% of the overall malaria infections in Honduras, the results reported here suggest that there is a dramatic increase in falciparum malaria in the area of Honduras investigated in this study. PMID- 9734228 TI - [Determination of selected mycotoxins in food. I. Selection of optimal conditions for the determination of aflatoxin M1 in milk using high-performance liquid chromatography methods]. AB - The aim of this study was to perform a optimized method for determination of aflatoxin M1 in milk. The manner of extraction and clean-up of milk extracts as well conditions of reaction of aflatoxin M1 with TFA and HPLC was described. The main steps of optimized method were: extraction of samples with chloroform, clean up of extracts on SPE C18 columns and by means of extraction with n-hexane, derivatisation of aflatoxin M1 with TFA (60 degrees C, 6 minutes) to acetal form- aflatoxin M2a and determination of aflatoxin by means of the RP-HPLC technique. The mobile phase was a mixture of methanol, isopropanol and water (18 + 7 + 75). Fluorometric detection was made at 370/418-700 nm. The mean recovery of aflatoxin M1 dependent on fortification level was 62-67%, limit of detection was 0.01 microgram/1 of milk. PMID- 9734229 TI - [Determination of selected mycotoxins in food. II. Selection of optimal conditions for the determination of fumonisin B1 and B2 in corn products using high performance liquid chromatography]. AB - The aim of this study was to perform a optimised method for determination of fumonisins B1 and B2 in corn products. The manner of extraction and clean-up of corn products extracts as well conditions of reaction of fumonisins with OPA was described. The main steps of optimised analytical procedure were: extraction of sample with methanol and water (3 + 1), clean-up of extracts on SAX column, derivatisation with OPA, and determination by means of RP-HPLC. The mobile phase was a mixture of methanol, water and acetic acid (75 + 24 + 1). Fluorometric detection was made at 370/440 nm. The mean recovery of fumonisins dependent on fortification level and product was 64-95%, limit of detection for each of fumonisins was 15 micrograms/kg. PMID- 9734230 TI - [Studies for measuring nitrates and nitrites in food served to patients in hospitals from the province of Bialystok]. AB - The aim of this study was determination of daily level intake of nitrates and nitrites in the individual meals in the daily food rations of patients from the hospitals from the province of Bialystok. The studied material were meals and total diets, with regard to selected food products, which were taken to preparing the dishes. The investigation materials were taken from February to June 1996. Analysis of nitrates and nitrites contents was realized by the commonly method based on the reaction of Griess. Nitrate was reduced to nitrite on a cadmium column whereupon, it was determined colorimetrically. The daily dose of nitrates for patients of 70 kg weight to the level on 350 mg KNO3/person and nitrites on 14 mg NaNO2/person, was established according to the Expert Committee FAO/WHO recommendations that acceptable daily intake level of nitrates on 5 mg and nitrites--0.2 mg to 1 kg body weight. In the light of these investigations it was found that main nitrates source in daily food rations were dinners containing red beet, while the great amount of nitrates was supplied in breakfasts and suppers with rennet maturing cheeses and sausages. From among all evaluated daily diets, 1/3 exceeded the admissible daily intake levels of nitrates and nitrites. PMID- 9734231 TI - [Molecular mechanisms of chemically induced carcinogenesis]. AB - In this review recent point of view concerning the molecular mechanisms of chemically induced carcinogenesis is presented. The new and promising trends of neoplasia investigations are based on discovery of protooncogenes and tumor suppressor genes, which maintain tissue homeostasis by controlling cellular proliferation and differentiation. It is generally recognised, that mutations induced by genotoxic carcinogens, particularly those resulting in activation of protooncogenes and inactivation of suppressor genes, play a crucial role in the initiation step of multistage process of tumorigenesis. Tumor promotion is recognized as a process whereby initiated cells are stimulated to selective growth and then, to develop into the cancer during progression step. Tumor promotion can be affected by many nongenotoxic carcinogens. In this review the attention is given to the mutational activation of the c-ras oncogenes and inactivation of p53 suppressor gene in rodent and human cancers by genotoxic carcinogens. Moreover, the significance of nongenotoxic carcinogens and the mechanisms by which these compounds may accelerate tumorigenesis are discussed. PMID- 9734232 TI - [The effect of nuarimol on the mutagenic activity of n-nitrosodimethylamine and 2 acetylaminofluorene in mouse erythrocytes]. AB - Nuarimol, the structural analogue of DDT, similarly to other polychlorinated aromatic hydrocarbons, induces monoxygenase activity. N-nitrosodimethylamine (NDMA) and 2-acetylaminofluorene (2-AAF) belong to chemical compounds exhibiting strong mutagenic and carcinogenic properties followed the metabolic activation. Genotoxic activity of promutagens, including NDMA and 2-AAF depends on the activity of monoxygenase enzymes. The study aimed at clarification of the effect of nuarimol on the mutagenic activity of NDMA and 2-AFF in in vivo micronucleus test. The experiments were performed on Swiss mice, which were exposed to nuarimol or Aroclor 1254 (as the positive control) followed by the exposure to NDMA or 2-AAF. The micronuclei were counted in the bone marrow polychromatic erythrocytes and in the erythrocytes of the peripheral blood. The results show that NDMA as well as 2-AAF induce failures in the genetic material in the bone marrow cells. Nuarimol given to the mice before the exposure to NDMA or 2-AAF did not cause changes in the micronuclei frequency. However, the prior intoxication by Aroclor 1254 resulted in the increase of the number of erythrocytes with micronuclei induced by NDMA in bone marrow and peripheral blood. This effect has not been observed in the mice intoxicated by 2-AAF prior to Aroclor 1254. PMID- 9734233 TI - Evaluation of effect of Metarhizium anisopliae on reduction in the number of Blattella germanica L. AB - Estimates were given of effectiveness of infestation of B. germanica L. By M. anisopliae strains originating from different regions of Poland. The investigated strains caused high mortality among experimental insects. Especially effective in reducing numbers of B. germanica were strains Browsk and Pruszyn, which even at a low density of spores caused high mortality of the test insects. PMID- 9734234 TI - [Sensitivity to disinfectants of Candid albicans strains isolated from the hospital environment]. AB - In recent years an increase of the incidence of Candida infections caused mainly by C. albicans strains especially in high risk inpatients with neoplasms, decreased immunity, burns and after treatment with multiple antibiotics has been observed. Candida organisms are particularly dangerous for newborns being responsible for about 30% of septicaemia cases in newborns in intensive care units. Fungal infections can be endogenous in origin but exogenous infection sources occur in hospitals. The cause of the latter are errors in aseptic management and insufficiently disinfected medical instruments and equipment. The purpose of the study was a comparison of the sensitivity to disinfectants of C. albicans belonging to two laboratory strains C. albicans PZH and C. albicans ATCC 10231 used for the determination of concentrations of two disinfectants used. Besides that, this sensitivity was determined in 14 strains isolated from the patients and one from the circuit of dialysis solution supply to artificial kidney. The study was carried out by the qualitative suspension method, in which the cells in the fluid were subjected to the action of disinfectants, and by the carrier method in which the cells of the microorganisms were present on the surface of metal cylinders. By the suspension method the sensitivity was determined to chloramine T in concentrations from 5.0% to 0.001%, formalin from 10.0% to 0.25%, glutaraldehyde from 2.0% to 0.1%, Septyl from 3.5% to 0.25%. The exposure time was 5, 10, 15, 30 and 60 minutes. The tested strains differed in their sensitivity to the disinfectants used. The greatest interstrain differences were observed in the sensitivity to the disinfectants used. The greatest interstrain differences were observed in the sensitivity to chloramine T. The highest concentrations were tolerated by the strains isolated from the patients and from the artificial kidney circuit as well as by the standard strain ATCC 10231. In the 10-minute exposure time accepted by us as comparison standard these strains were 200-time less susceptible to chloramine than the standard C. albicans PZH strain. Two strain isolated from the patients were tenfold as sensitivive. The sensitivity to the remaining tested disinfectants showed less evident differences. The sensitivity of the strains from the patients to formalin was similar to that of the standard PZH strain. A similar sensitivity was found to Septyl, with the exception of the strain from the artificial kidney circuit which was sevenfold less sensitive than the PZH strain. In the case of glutaraldehyde 9 strains from the patients and the ATCC 10231 strain were two or four times less sensitive than the PZH strain. No cross-sensitivity or tolerance to the disinfectants were noted in the study. Both standard strains were similarly sensitive to formalin, but the ATCC 10231 strain was less sensitive to Septyl, glutaraldehyde and chloramine T. In the experiment by the carrier method the effect was evidenced of the surface on the action of disinfectants. This was particularly evident in the case of chloramine T. Even in sensitive strains the disinfection parameters (concentration and exposure time) were significantly higher than in the suspension method. The least sensitive strains survived the effect of 5% chloramine during 2 hours of exposure. Septyl in the working concentration 2.5% at 10-minute exposure time disinfected all carriers with the exception of that carrying the strain isolated from the artificial kidney circuit, which survived 15% Septyl exposure during 10 minutes. The disinfectant Aldesan (2% glutaraldehyde) and formalin 8% killed all fungi during 10 minutes. The study shows that the sensitivity of C. albicans strains to disinfectants varies. For the assessment of the fungicidal action of disinfectants the standard test ATCC 10231 should be used since its sensitivity was similar to that of most strains from the patients and medical equipment. (ABSTRACT TRUNCATED) PMID- 9734235 TI - [Use of Chapman and Baird-Parker media for isolation of Staphylococci from swimming pool water]. PMID- 9734236 TI - [Hygienic assessment of conditions for education and recreation of students in children's homes]. AB - The study was carried out in 1995 in all children's homes listed in the sanitary epidemiological stations in the country, with 16,471 inmates, among them 13,778 were attending elementary and secondary schools. The conditions for education and recreation in these homes were studied by the workers of the children and adolescent sections of the sanitary-epidemiological stations overseeing children's homes in the area of their coverage. For the study a uniform questionnaire was used. On the basis of the results the conclusion is put forward, that the conditions of education and recreation of children differ considerably from one home to another. In most homes the conditions for education and recreation are good. In certain homes short-comings were found, e.g. insufficient lighting of working places, which require actions for their elimination to avoid possible consequences for health. The actions undertaken for sending in 1995 of a possibly high number of inmates to summer or winter camps were brought into realization in a considerable degree. In summer 71% and in winter 23% of the inmates attending schools participated in various forms of organized recreation. PMID- 9734237 TI - [A century of investigations and work connected with the potable water supply for Krakow]. AB - The aim of the paper was to present the works and results of investigations connected with the control of water quality for the inhabitants of Krakow city. The investigations started in 1898 and were performed by K. Olszewski, being advised by O. Bujwid. The water--works started to operate in 1901, and ground water coming from several wells dug to create the water intake system. The increase of potable water demands of growing Krakow, and unacceptable pollution level of Vistula water decided to base the water intake for Krakow on rivers: Rudawa, Sanka and Dlubnia, but since 1986 also on Raba river. PMID- 9734238 TI - [Report on scholarships in the BIBRA International (England) and from the Biology Department of the University in Padua (Italy)]. PMID- 9734239 TI - Bromelain. PMID- 9734240 TI - Boswellia serrata. PMID- 9734241 TI - alpha-Lipoic acid. PMID- 9734242 TI - Do patients really use their cholesterol medicines? PMID- 9734243 TI - Smart estrogens may decrease heart-attack risk. PMID- 9734245 TI - Garlic oil: no impact on lipids. PMID- 9734244 TI - Advances in angioplasty: beyond balloons. PMID- 9734246 TI - Diuretics: first-line treatment for hypertension in the elderly. PMID- 9734248 TI - I lie awake almost every night feeling my heart skip beats. My doctor tells me not to worry, but I can't help being concerned. What can I do about this problem? PMID- 9734247 TI - Long-term exercise a better bet to protect arteries. PMID- 9734249 TI - I am 58 years old and told my doctor I wanted to start an exercise program. I have never had any chest pain, but she told me I should have an exercise test because I have hypertension and diabetes. I felt fine during the test, but after four minutes they stopped me and told me my results were very abnormal. They said my EKG showed 3 mm of change. Two days later, I had a coronary angiogram, and two days after that I had bypass surgery! Was all this necessary? PMID- 9734250 TI - I take Zocor for high cholesterol. My doctor checks my lipid levels every six months and they have been pretty good for a few years now. But he also checks my liver-function tests, because liver damage is one of the side effects of this drug. How worried should I be about this problem, and is checking my liver tests every six months often enough? PMID- 9734251 TI - What causes cancer? PMID- 9734253 TI - The hostile heart. PMID- 9734254 TI - I'm a 44-year-old former smoker. I have just been diagnosed with colitis, and I've heard that smoking can help my condition. There is no cancer or heart disease in my family. Should I start smoking again? PMID- 9734252 TI - Dog bites man. PMID- 9734256 TI - Tailoring hypertension treatment. PMID- 9734255 TI - Hormones and mood. PMID- 9734257 TI - Hip health. PMID- 9734258 TI - Secondhand smoke. PMID- 9734259 TI - New HPV test for cervical cancer. PMID- 9734260 TI - New studies question tamoxifen's benefits. PMID- 9734261 TI - I am 59 years old and recently had a complete hysterectomy for endometrial cancer. The pathology report showed no migration of cancer cells outside the endometrium. Is it safe for me to take estrogen as hormone replacement therapy? PMID- 9734263 TI - In the June 1998 issue of HWHW, you wrote that a 3.3 ounce T-bone steak, broiled, yields 26.8 grams of protein. Since 3.3 ounces weighs roughly 100 grams, I'm wondering what makes up the outstanding grams in that T-bone steak. Is it fat? PMID- 9734264 TI - 23rd National Cancer Congress of the German Cancer Society. Berlin, 8-12 June 1998. Abstracts. PMID- 9734262 TI - I recently had a hysterectomy in which my cervix was not removed. What is the correct hormone replacement therapy for me? Do I have to take progesterone? Will the estrogen increase my risk of cervical cancer? PMID- 9734265 TI - American Association of Oral and Maxillofacial Surgeons (AAOMS) 80th annual meeting. New Orleans, Louisiana, USA. September 16-20, 1998. Abstracts. PMID- 9734266 TI - 30 years ago in Clinical Toxicology. PMID- 9734267 TI - Looking back. PMID- 9734268 TI - The American Academy of Clinical Toxicology: a historic pediatric perspective. PMID- 9734269 TI - Roots and circles in medical toxicology: a personal reminiscence. PMID- 9734270 TI - Annual meeting of the American Academy of Otolaryngology--Head and Neck Surgery Foundation, Inc., San Antonio, Texas, USA. September 13-16, 1998. Abstracts. PMID- 9734271 TI - International Congress of Toxicology--VIII, Paris, France. 5-9 July 1998. Abstracts. PMID- 9734272 TI - Use of solid phase extraction disks for the GC-MS analysis of acidic and neutral herbicides in drinking water. AB - An analytical procedure was developed in which thirteen herbicides (10 acidic and 3 neutral compounds) may be extracted from drinking water samples using solid phase extraction disks and subsequently analyzed by gas chromatography-mass spectrometry. Using 8 replicate samples consisting of reagent water fortified with nanogram quantities of each herbicide, the average percent recoveries and method detection limits were determined for each analyte. The method was found to be suitable for the determination of individual herbicides in drinking water at concentrations of approximately 10 ng/L. PMID- 9734273 TI - A fish hepatoma cell line (PLHC-1) as a tool to study cytotoxicity and CYP1A induction properties of cellulose and wood chip extracts. AB - Cytotoxicity and CYP1A induction properties of celluloses and wood chips were studied with a teleost liver cell line, PLHC-1. Cells were exposed to acetone extracts of celluloses produced using new bleaching techniques (elemental chlorine free, ECF; totally chlorine free, TCF) in two sulphate mills or without any bleaching (unbleached, UB) in a sulphite mill. In another set of exposures, celluloses (ECF and TCF bleached) and wood chips (from pine and birch) were collected from a sulphate mill, extracted with acetone, and the extracts used to treat the cells. After exposure, O-deethylation of 7-ethoxyresorufin (EROD, a measure of cytochrome P4501A (CYP1A) catalytic activity), and total protein content, a measure of cytotoxicity, were assayed. The presence of the CYP1A protein in the exposed cells was assessed by immunoblotting. The cellulose and wood chip extracts were able to cause both cytotoxicity and EROD induction in the PLHC-1 cells. In the exposures conducted with the material from three different mills, the celluloses made of birch were more cytotoxic and more potent inducers of EROD activity than were the celluloses of pine. Further, UB celluloses increased EROD activity and caused cytotoxicity at lower doses than material bleached with modern bleaching techniques. In the exposures made with material from one single mill, there were no clear trends between the celluloses made of pine or birch. Wood chips of pine, however, were more cytotoxic than wood chips of birch. Especially with pine wood chips, cytotoxicity interfered with the induction of EROD activity, thus complicating the evaluation of CYP1A induction. CYP1A protein content was not detected in cells exposed to extracts of celluloses or wood chips, possibly due to low amounts of protein available for the assay. Wood and pulp processing, like bleaching, may change the chemical composition of the raw material in a way that reduces the potency for biological effects of the final product, cellulose. This could explain why both UB celluloses and wood chips were more potent in the cells than ECF or TCF bleached celluloses. In this study the PLHC-1 cell line showed its potential for use in evaluating the biological activity existing in pulp and paper mill products and raw materials. The identity and source of the compounds that were able to affect the PLHC-1 cell line remain to be determined. PMID- 9734274 TI - Pyrene degradation by Mycobacterium sp. strain KR2. AB - A Mycobacterium sp., strain KR2 which was able to utilise pyrene as sole source of carbon and energy was isolated from a polycyclic aromatic hydrocarbon (PAH) contaminated soil originating from the area of a former gaswork plant. The isolate metabolised up to 60% of the pyrene added (0.5 mg/mL) within 8 days at 20 degrees C. Cis-4,5-pyrene dihydrodiol, 4,5-phenanthrene dicarboxylic acid, 1 hydroxy-2-naphthoic acid, 2-carboxybenzaldehyde, phthalic acid, and protocatechuic acid were identified as degradation products. Based on these findings a degradation pathway for pyrene is suggested which is in good accordance with the data published so far on bacterial pyrene metabolism. PMID- 9734275 TI - Estimating dermal transfer from PCB-contaminated porous surfaces. AB - Health risks posed by dermal contact with PCB-contaminated porous surfaces have not been directly demonstrated and are difficult to estimate indirectly. Surface contamination by organic compounds is commonly assessed by collecting wipe samples with hexane as the solvent. However, for porous surfaces, hexane wipe characterization is of limited direct use when estimating potential human exposure. Particularly for porous surfaces, the relationship between the amount of organic material collected by hexane and the amount actually picked up by, for example, a person's hand touch is unknown. To better mimic PCB pickup by casual hand contact with contaminated concrete surfaces, we used alternate solvents and wipe application methods that more closely mimic casual dermal contact. Our sampling results were compared to PCB pickup using hexane-wetted wipes and the standard rubbing protocol. Dry and oil-wetted samples, applied without rubbing, picked up less than 1% of the PCBs picked up by the standard hexane procedure; with rubbing, they picked up about 2%. Without rubbing, saline-wetted wipes picked up 2.5%; with rubbing, they picked up about 12%. While the nature of dermal contact with a contaminated surface cannot be perfectly reproduced with a wipe sample, our results with alternate wiping solvents and rubbing methods more closely mimic hand contact than the standard hexane wipe protocol. The relative pickup estimates presented in this paper can be used in conjunction with site specific PCB hexane wipe results to estimate dermal pickup rates at sites with PCB-contaminated concrete. PMID- 9734277 TI - Israel Diabetes Association annual meeting. 27 January 1998. Abstracts. PMID- 9734276 TI - Alterations of signal transduction pathways involved in 2,3,7,8 tetrachlorodibenzo-p-dioxin-induced malignant transformation of human cells in culture. AB - Effects of signal transduction pathways in TCDD-induced neoplastic transformation of human cells were assessed with respect to PLC-coupled signaling pathways, adenylyl cyclase-mediated responses and PKC isozyme expressions. A lower stimulation of the intracellular free calcium levels with exposure to extracellular ATP or histamine was observed in the transformed cells, as compared to the parental cells. While the steady-state level of IP3 was higher in the transformed cells, the magnitude of stimulation of IP3 generation by ATP or histamine was significantly lower in the transformed cells than the parental cells. These results indicate that a downregulation PLC-coupled signaling pathways may be involved in the TCDD-induced transformation of human cells. While the steady-state levels of cAMP accumulation were similar between the two cell lines, treatment of PGE2, a potent differentiation inducer, stimulated a higher accumulation of cAMP in the parental cells but isoproterenol, a typical beta adrenergic agonist, did not induce a significant difference between the two cell lines. These results suggest that desensitization of cAMP-mediated response to extracellular signals including differentiation signals may be associated with a possible mechanism of the carcinogenesis. Elevated expression of PKC-alpha, gamma, -zeta, -epsilon, -lambda, and -tau were observed in TCDD-transformed cells, indicating a possible association of altered expression of PKC isozymes with TCDD-induced transformation of human cells. The present study demonstrates that alterations of signal transduction pathways are involved in the TCDD-induced transformation of human cells and provides a valuable basis to investigate effects of signaling pathway as a possible mechanism of TCDD-induced carcinogenesis in human cells. PMID- 9734278 TI - 21st Annual meeting of the American Society of Primatologists. June 28-July 2, 1998. Abstracts. PMID- 9734279 TI - 33rd Meeting of the Canadian Congress of Neurological Sciences. Montreal, Quebec, Canada. June 16-20, 1998. Abstracts. PMID- 9734280 TI - 19th Annual meeting of the Society for Clinical Trials. Atlanta, Georgia, USA. May 17-20, 1998. Abstracts. PMID- 9734281 TI - European Society for Clinical Investigation, 32rd meeting. Cracow, Poland, 16-19 April 1998. Abstracts. PMID- 9734284 TI - 5th International Conference on Systemic Lupus Erythematosus. Cancun, Mexico, April 20-25, 1998. Abstracts. PMID- 9734283 TI - 16th International Congress on Acoustics and 135th meeting of the Acoustical Society of America. Seattle, Washington, USA. 20-26 June 1998. Abstracts. PMID- 9734282 TI - 3rd International Conference on Dietary Assessment Methods. Arnhem, The Netherlands, 6-9 May 1998. Abstracts. PMID- 9734286 TI - German Society for Experimental and Clinical Pharmacology and Toxicology, 39th spring meeting. Mainz, Germany, 17-19 March 1998. Abstracts. PMID- 9734285 TI - Multiple Sclerosis: Frontiers in Science and Patient Care and Disease Management. Birmingham, United Kingdom, 5-6 May 1998. Abstracts. PMID- 9734287 TI - 1998 Annual meeting and 38th Symposia of the Japan Society of Plant Physiologists. May 3-5, 1998. Abstracts. PMID- 9734289 TI - Annual meeting of the Swiss Society of Cardiology and the Swiss Association for the Prevention of Hypertension. Interlaken, 14-16 May 1998. Abstracts. PMID- 9734288 TI - American Society of Regional Anesthesia 23rd annual meeting. Seattle, Washington, USA. May 14-17, 1998. Abstracts. PMID- 9734290 TI - 21st Annual Conference on Shock. San Antonio, Texas, USA. June 14-17, 1998. Abstracts. PMID- 9734291 TI - Drug resistance in malaria parasites of animals and man. PMID- 9734292 TI - Molecular pathobiology and antigenic variation of Pneumocystis carinii. PMID- 9734293 TI - Ascariasis in China. PMID- 9734294 TI - The generation and expression of immunity to Trichinella spiralis in laboratory rodents. PMID- 9734295 TI - Population biology of parasitic nematodes: applications of genetic markers. PMID- 9734296 TI - Schistosomiasis in cattle. PMID- 9734297 TI - The effect of stimulus omission and interstimulus interval (ISI) on electrodermal habituation at short ISIs. AB - An experiment was conducted to examine the effects of complete stimulus omission and interstimulus interval (ISI) variability on habituation of the skin conductance response. A new scoring technique allowed shorter ISIs to be used (0.5-1.5 s) than in previous studies. It was found that both stimulus omission and variation of a previously constant ISI had a significant effect on the course of habituation at short ISIs. The results are consistent with comparator models of habituation and suggest that participants encode the ISI of a series of stimuli presented as short ISIs. PMID- 9734298 TI - DSM-III-R and ICD-10 personality disorder features among women experiencing two types of self-reported homesickness: an exploratory study. AB - This study investigated the intensity of DSM-III-R and ICD-10 personality disorder features among females experiencing self-reported homesickness. Three groups were compared: (a) a group of women experiencing chronic feelings of homesickness (CHS); (b) a group of women experiencing episodic attacks of homesickness, each time they go on holidays (EHS); and (c) a group of healthy control females, recruited from the general population (HC). This study aimed to investigate whether the homesick participants showed stronger features of personality pathology than the controls and whether those who report experiencing chronic feelings of homesickness showed stronger features of personality pathology than those who at the moment of testing were not in an actual state of homesickness. Glass effect sizes revealed that the DSM-III-R avoidant and dependent and the ICD-10 anxious and dependent traits were most strongly associated with either type of homesickness. Finally, CHS was particularly associated with passive-aggressive traits and EHS with sadistic traits. Based on the stories of 21 homesick women, a link between adverse attachment experiences in childhood and certain personality features on the one hand and homesickness on the other is tentatively suggested. PMID- 9734299 TI - The role of social deviance and violations in predicting road traffic accidents in a sample of young offenders. AB - Lawton, Parker, Stradling & Manstead (1997) examined the relationship between mild social deviance (West, Elander & French, 1993 a), driving violations and road traffic accident involvement in a sample of 830 drivers. The relationship between mild social deviance and accident involvement was shown to be partly mediated by propensity to commit driving violations and by factors associated with driver age. The present research replicates and extends this study with a sample of 100 young, male offenders. Self-reports of violations and errors (using the Manchester Driver Behaviour Questionnaire), an extended measures of social deviance, speed preference and accident involvement were collected, together with information about age, annual mileage driven and the type of offence for which the driver was under remand. Factor analysis of the social deviance items yielded two factors: extreme and mild social deviance. Logistic regression was used to investigate the relationship between predictor variables and accident involvement. Both propensity to commit driving violations and extreme social deviance predicted accident involvement in this sample. However, the relationship between extreme social deviance and accident involvement was partly mediated by a tendency to commit driving violations. The implications of the findings for intervention strategies aimed at the prevention of accidents are considered. PMID- 9734300 TI - Can animals detect when their owners are returning home? An experimental test of the 'psychic pet' phenomenon. AB - In his book, Seven Experiments That Could Change The World, Rupert Sheldrake suggested that the public carry out experiments to test whether pets can psychically detect when their owners are returning home. The first of these tests was undertaken by an Austrian television company and involved an owner in the north-west of England, Pam Smart (PS) and her dog (Jaytee). The test appeared remarkably successful and seemed to show Jaytee responding when PS set off to return home from a remote location. Rupert Sheldrake and PS asked the authors if they would like to carry out their own investigation into Jaytee's abilities. This paper outlines various 'normal' explanations that might account for the phenomenon and presents an experimental design that minimizes these possibilities. The paper then details the procedure and results of four experiments. Analysis of the data did not support the hypothesis that Jaytee could psychically detect when his owner was returning home. Finally, the paper discusses a possible reason for the difference in results of these studies and those carried out by the Austrian television company. PMID- 9734301 TI - Development of spatial memory and spatial orientation in preschoolers and primary school children. AB - The present study addresses the question of what kind of information children use when orientating in new environments, if given proximal and distal landmarks, and how spatial memory develops in the investigated age groups. Ten 5-year-old, ten 7 year-old and ten 10-year-old children were presented with the 'Kiel Locomotor Maze', containing features of the Radial Arm Maze and the Morris Water Maze, in order to assess spatial memory and orientation. Children had to learn to approach baited locations only. Task difficulty was equated with respect to the children's age. Training was given until the children reached criterion. During testing, the maze configuration and response requirements were systematically altered, including response rotation, cue rotation, cue deletion and response rotation with cue deletion in order to assess the spatial strategies used by the children. During training and testing, working-memory errors (WM), reference-memory errors (RM) and working-reference memory errors (WR) were recorded. As expected, no difference between age groups appeared during training, thus confirming comparable task difficulty across age groups. During testing, age groups differed significantly with regard to the orientation strategy used. The 5-year-olds were bound to a cue strategy, orientating towards local, proximal cues. The 10-year olds mastered all tasks, thus displaying a place strategy, being able to use distal cues for orientation, and were even able to do so after being rotated 180 degrees. The 7-year-olds proved to be at an age of transition: five of them were bound to a cue strategy, five children were able to adopt a place strategy. The differences in the orientation strategies used by children of different age groups was reflected by the sum of errors they made, also by RM. WM were found to be rare, especially in older children. We conclude that preschoolers use a cue strategy, that the development of place strategies occurs during primary school age and seems to be complete by the age of 10 years. PMID- 9734302 TI - Fungicidal properties, sterol binding, and proteolytic resistance of the synthetic peptide D4E1. AB - The fungicidal properties of the synthetic peptide D4E1 were studied with nongerminated and germinating conidia of Aspergillus flavus, Aspergillus fumigatus, Aspergillus niger, Fusarium moniliforme, and Fusarium oxysporum. The minimal lethal concentrations (MLC) needed to kill 100% of germinating conidia of A. fumigatus, A. flavus, and A. niger were 12.5, 12.5, and 25 microM, respectively. The MLC value for nongerminated and germinating conidia of both Fusarium spp. was 3.0 microM. Except for A. fumigatus, D4E1 was inactive against the nongerminated conidia of the Aspergillus spp. Physicochemical studies showed D4E1 complexed with ergosterol, a sterol present in conidial walls. Cholesterol, present in nongerminated conidia of F. moniliforme, had a greater affinity for D4E1 than did ergosterol. D4E1 was more resistant to fungal and plant protease degradation than the natural peptide, cecropin A. These in vitro results suggest D4E1 is a candidate for transgenic expression in plants to enhance host resistance to fungal infection. PMID- 9734303 TI - Evidence that a deferrioxamine B degrading enzyme is a serine protease. AB - Siderophores are organic biomolecules synthesized by a wide variety of microbes. The molecules sequester ferric ion from environments where it is present at extremely low concentrations. Siderophores are of consequence with respect to microbial nutrition, pathogenicity, virulence, and microbe-plant interactions. How siderophores are degraded and returned to the carbon and nitrogen cycles is not well understood. The catalytic activity of an enzyme from a bacterium that degrades the siderophore deferrioxamine B has been examined. While the degradation of deferrioxamine B is sensitive to sulfhydryl and metal moiety inhibitors, the data presented is most consistent with the hypothesis that the enzyme uses a hydroxyl moiety (serine peptidase) to catalyze the degradation of deferrioxamine B. If sulfhydryl and metal inhibitors are simultaneously present at concentrations that when alone only partially inhibit the enzyme, the enzyme is unable to catalyze deferrioxamine B dissimilation. Analysis of the inhibitor experiments conducted led to the conclusion that the deferrioxamine B degrading enzyme is a serine-peptidase-like enzyme that needs calcium ions and sulfhydryl groups to be fully activated or stabilized. The knowledge of the catalytic moieties of the enzyme will be exploited to purify the enzyme. PMID- 9734304 TI - Molecular analysis of bacterial isolates and total community DNA from kraft pulp mill effluent treatment systems. AB - Chloroaliphatics are major components of bleached kraft mill effluents. Gene probes and oligonucleotide primers were developed to monitor kraft pulp mill effluent treatment systems for the presence of key genes (dehalogenases) responsible for the dehalogenation of chloroaliphatic organics. The primers were used for polymerase chain reaction (PCR) analysis of genomic DNA extracted from dehalogenating bacterial isolates and from total community DNA extracted from water and sediments of mill effluent treatment system. PCR amplification with oligonucleotide primers designed from dhlB, encoding the haloacid dehalogenase from Xanthobacter autotrophicus, revealed the presence of dehalogenase genes in both aerated lagoons and stabilization basins. Similarly, positive results were obtained with mmoX primers designed from the soluble methane monooxygenase gene of Methylococcus capsulatus Bath. The haloacetate dehalogenase encoding gene (dehH2) from Moraxella sp. was typically not detected in mill effluent treatment systems unless the biomass was selectively enriched. DNA sequence analysis of several PCR fragaments revealed significant similarity to known dehalogenase amd methane monooxygenase genes. The results indicated a broad distribution of known dehalogenation genes and bacteria with chloroorganic-degrading potential in the mill effluent treatment systems. PMID- 9734305 TI - The phbC (poly-beta-hydroxybutyrate synthase) gene of Rhizobium (Sinorhizobium) meliloti and characterization of phbC mutants. AB - Defined insertion mutations have been constructed in the Rhizobium (Sinorhizobium) meliloti phbC gene, which encodes poly-beta-hydroxybutyrate (PHB) synthase. The locus was isolated and subcloned from a genomic library of R. meliloti Rm1021 by complementation of phbC mutation of Alcaligenes eutrophus. PHB production was detected in wild-type R. meliloti under nutrient-limited conditions but not in rich medium. No PHB production was detected in the R. meliloti phbC mutants. The DNA sequence of the R. meliloti phbC gene was determined. The deduced polypeptide sequence is homologous to previously identified PhbCs from other bacteria. The R. meliloti phbC locus maps to pRmeSU47a, the smaller of the two megaplasmids in this strain. PMID- 9734306 TI - Identification of sigma 32-like factors and ftsX-rpoH gene arrangements in enteric bacteria. AB - Western blot analyses using anti-Escherichia coli K-12 sigma 32 antibodies and Southern blot analyses using rpoH and ftsX DNA probes were performed using different enteric bacteria. Results show that the bacterial strains analysed have sigma 32-like transcription factors and ftsX and rpoH homologs in a similar map position. Although the presence of sigma 32-like factors seems to be extended to all Proteobacteria, rpoH and ftsX homologs seem to be present as neighbors in the genome only in the enteric bacteria. PMID- 9734307 TI - Double-stranded RNA mycoviruses in species of Aspergillus sections Circumdati and Fumigati. AB - Isolates (178) belonging to Aspergillus sections Fumigati, Candidi, Clavati, and Circumdati were tested for the presence of double-stranded RNA (dsRNA) genomes. Altogether, 5.6% of the Aspergillus strains examined were infected with dsRNAs. dsRNA segments indicative of mycovirus infection were observed for the first time in Neosartorya hiratsukae, Neosartorya quadricincta, Petromyces alliaceus, and Aspergillus clavatus strains. Correlation was not observed between ochratoxin production and dsRNA content of the strains. This is the first report on the detection of naturally occurring dsRNAs in Aspergillus species that are able to reproduce sexually. The detection of dsRNA in sexual aspergilli gave us a chance to examine the transmission of these segments through ascospores. A Neosartorya hiratsukae strain transmitted the dsRNAs efficiently through sexual spores, while the stromata embedding the asci in Petromyces alliaceus did not transmit one of the dsRNA segments. The 0.6-kb dsRNA segment that was present in the single stromatal cultures was found to be located in the mitochondrial fraction of this strain. This observation indicates that some mechanisms exist in aspergilli to exclude cytoplasmically located dsRNA molecules from stromatal structures. PMID- 9734308 TI - Kinetic analyses of Biolog community profiles to detect changes in inoculum density and species diversity of river bacterial communities. AB - The kinetics of response curves from Biolog community profiles for heterotrophic bacteria from a river in Nova Scotia, Canada have been analyzed to generate lag, slope, and asymptote parameters. The river water samples were treated with one of three supplements of Escherichia coli (in situ levels, 10(3) CFU/mL, or 10(6) CFU/mL) and one of five concentrations of chlorine (0, 1, 3, 5, or 7 ppm) to satisfy a full factorial design. The chlorine treatments decreased the inoculum density by up to 2 log values and decreased the species evenness. The E. coli supplements increased the inoculum density and decreased the species richness. Examination of the asymptotes did not reveal any significant effects owing to E. coli, but differences owing to the chlorine were detected. Analyses of the slopes showed a similar insignificance of the effects of E. coli and a lack of treatment effect owing to chlorine. The lag analyses also showed no significant E. coli effects, but showed a significant effect owing to chlorine. The discrepancy produced with the slope analysis (i.e., no chlorine effect) may represent an anomaly of the Biolog community approach. The use of lag phase was impaired because of the problem of infinite lags from wells that had no response, but a principle component analysis with a reduced set of substrates did suggest some influence of E. coli on the community profile. An examination of the substrates metabolized by the river water compared with pure E. coli revealed that the Biolog profiles of the river communities were not a simple summation of the component parts. In light of the lack of uniformity between these analyses, where the outcome depended on which parameter was used, caution is advised in interpreting Biolog community profiles on the basis of only one parameter. PMID- 9734309 TI - Incidence of enteroviruses in Mamala Bay, Hawaii using cell culture and direct polymerase chain reaction methodologies. AB - The consequence of point and nonpoint pollution sources, discharged into marine waters, on public recreational beaches in Mamala Bay, Hawaii was evaluated using virus cell culture and direct reverse transcriptase-polymerase chain reaction (RT PCR). Twelve sites, nine marine, two freshwater (one stream and one canal), and one sewage, were assessed either quarterly or monthly for 1 year to detect the presence of human enteric viruses. Water samples were concentrated from initial volumes of 400 L to final volumes of 30 mL using Filterite electronegative cartridge filters and a modified beef extract elution procedure. Cell culture was applied using the Buffalo Green Monkey kidney cell line to analyze samples for enteroviruses. Positive samples were also evaluated by RT-PCR, using enterovirus specific primers. Levels of RT-PCR inhibition varied with each concentrated sample. Resin column purification increased PCR detection sensitivity by at least one order of magnitude in a variety of sewage outfall and recreational marine water samples but not in the freshwater canal samples. Using cell culture, viable enteroviruses were found in 50 and 17% of all outfall and canal samples, respectively. Samples were positive at beaches 8% of the time. These data illustrate the potential public health hazard associated with recreational waters. Using direct PCR, viruses were detected at the outfall but were not found in any beach or canal samples, in part, owing to substances that inhibit PCR. Therefore, conventional cell culture is the most effective means of detecting low levels of infectious enteroviruses in environmental waters, whereas direct RT-PCR is rendered less effective by inhibitory compounds and low equivalent reaction volumes. PMID- 9734310 TI - Studies towards the synthesis of the hypermodified nucleoside of rat liver phenylalanine transfer ribonucleic acid: improved synthesis of the base beta hydroxywybutine. AB - An improved synthesis of the key intermediates (3 and 8) for the synthesis of beta-hydroxywybutines [[R-(R*,S*)]- and [S-(R*,R*)]-4], the most probable structures for the minor base from rat liver tRNA(Phe), has been achieved by the Wittig reaction between 1-benzyl-7-formylwye (1) and the phosphorane derived from (R)-2-[(methoxycarbonyl)amino]-3-(triphenylphosphonio)propanoate (10), followed by methylation, OsO4 oxidation, and cyclocondensation with COCl2 in the presence of pyridine. The racemic forms of beta-hydroxywybutines [(R*,S*)- and (R*,R*)-4], which were required for the determination of the optical purity of [R-(R*,S*)]- and [S-(R*,R*)]-4 by means of chiral HPLC, were conveniently prepared through pyrolysis of the cyclic carbonate 3 followed by NaBH4 reduction and catalytic hydrogenolysis. The samples of [R-(R*,S*)]- and [S-(R*,R*)]-4 were thus shown to be optically pure. PMID- 9734311 TI - Pregnane glycosides, gymnepregosides A-F from the roots of Gymnema alternifolium. AB - The structural elucidation of six new related polyoxypregnane glycosides, gymnepregosides A (1), B (2), C (3), D (4), E (5) and F (6), together with two known compounds, from the roots of Gymnema alternifolium (Asclepiadaceae) was achieved through on a detailed study of 1H- and 13C-NMR spectral data and chemical means. The results obtained for new compounds, 1-6, show that they are (20S)-pregn-6-ene-3 beta,5 alpha,8 beta,12 beta,14 beta,17 beta,20-heptaol or sarcostin 3-O-glycosides, and all the sugars at C-3 are beta(1-->4)-linked. Some of them possessed benzoyl, cinnamoyl and tigloyl residues as the ester linkages located at C-12 and/or C-20 of the aglycon. PMID- 9734312 TI - Antibacterials and antimycotics: Part 1: Synthesis and activity of 2-pyrazoline derivatives. AB - A series of 3-styryl-1,5-diphenyl and 5-styryl-1,3-diphenyl 2-pyrazolines of different substitutions has been synthesized by condensation of substituted alpha,beta-unsaturated ketones with phenylhydrazine hydrochloride in presence of catalytic amount of concentrated HCl. Compounds in the 3-styryl series had OMe, NMe2, NO2, OH and isopropyl substituents and those in the 5-styryl series had OMe, NMe2 and NOs. The 3-styryl-1,5-diphenyl compounds showed little variation in antibacterial activity towards gram-positive and gram-negative bacteria in terms of geometric mean minimum inhibitory concentrations (MIC). The 4',4"-NMe2, 4',4" NO2 and 4',4"-OMe compounds were found to possess the highest activity in the series. The 5-styryl-1,3-diphenyl series showed lower activities than the 3 styryl series. The in vitro antimycotic activity of the 4',4"-OH and 2',2"-OH substituted compounds showed good activity than the other molecules in the two series. PMID- 9734313 TI - A new subculture and nematocidal assay using a species of diplogastridae. AB - A new subculture method and a novel microplate assay for nematocidal activity using a species of Diplogastridae have been developed. The assay gives results rapidly, with high sensitivity in 4 h, and indicated good correlation between action mechanism and the nematode shape when examining 15 known compounds, including the antiparasitic avermectin, antimalarial quinine, and the gamma-amino n-butyric acidA (GABAA) activated Cl- channel antagonist picrotoxinin. Thus new assay could be used as a primary screening method for new nematocidal compounds. PMID- 9734316 TI - Selective muscarinic antagonists. II. Synthesis and antimuscarinic properties of biphenylylcarbamate derivatives. AB - A novel series of biphenylylcarbamate derivatives were synthesized and evaluated for binding to M1, M2 and M3 receptors and for antimuscarinic activities. Receptor binding assays indicated that biphenyl-2-ylcarbamate derivatives had high affinities for M1 and M3 receptors and good selectivities for M3 receptor over M2 receptor, indicating that the biphenyl-2-yl group is a novel hydrophobic replacement for the benzhydryl group in the muscarinic antagonist field. In this series, quinuclidin-4-yl biphenyl-2-ylcarbamate monohydrochloride (8l, YM-46303) exhibited the highest affinities for M1 and M3 receptors, and selectivity for M3 over M2 receptor. Compared to oxybutynin, YM-46303 showed approximately ten times higher inhibitory activity on bladder pressure in reflexly-evoked rhythmic contraction, and about 5-fold greater selectivity for urinary bladder contraction against salivary secretion in rats. Moreover, selective antagonistic activity was also observed in vitro. Further evaluation of antimuscarinic effects on bradycardia and pressor in pithed rats, and on tremor in mice, showed that YM 46303 can be useful for the treatment of urinary urge incontinence as a bladder selective M3 antagonist with potent activities and fewer side effects. PMID- 9734314 TI - Synthesis and muscarinic activity of a series of quinolines and naphthalenes with a 1-azabicyclo[3.3.0]octane moiety. AB - In order to discover a medicine effective against Alzheimer's disease, we synthesized a series of quinoline derivatives having a characteristic 1 azabicyclo[3.3.0]octane amine ring, and performed pharmacological evaluation of them. Acetylcholine esterase inhibitory activities of these derivatives were unexpectedly weak. Tests for central nervous muscarinic cholinergic receptor binding affinity indicated that these compounds had higher affinities to muscarinic M1 receptors than to M2 receptors. A series of naphthalene derivatives substituted with the 1-azabicyclo[3.3.0]octane ring were also synthesized and muscarinic M1 and M2 receptor binding affinity determined. These compounds had much higher affinity for M1 receptors than the quinoline derivatives, and 1-[N-(1 azabicyclo[3.3.0]octan-5-yl)methyl-N-methylamino]-4-nitronaph tha lene showed the highest affinity and selectivity. The ability of this compound to improve cognitive function was assessed using the passive avoidance test in scopolamine induced mice. PMID- 9734315 TI - Selective muscarinic antagonists. I. Synthesis and antimuscarinic properties of 4 piperidyl benzhydrylcarbamate derivatives. AB - A series of 1-substituted-4-piperidyl benzhydrylcarbamate derivatives were synthesized and evaluated for binding affinity to M1, M2 and M3 receptors, and for antimuscarinic activities. Receptor binding assays indicated that 1-benzyl-4 piperidyl benzhydrylcarbamate derivatives showed higher affinities for M1 and M3 receptors, and good selectivities for M3 over M2 receptor, than the corresponding ester analog. These results indicate that the urethane bond is a novel linker for muscarinic antagonists, and serves to lock the molecular conformation and allows the hydrophobic portion and cationic site of the molecule to bind to M1 and M3 muscarinic receptors. Among the prepared compounds, 1-(4-methylaminobenzyl)-4 piperidyl benzhydrylcarbamate monohydrochloride (18b, YM-58790) exhibited potent inhibitory activity on bladder pressure in reflexly-evoked rhythmic contraction, comparable to oxybutynin and was approximately ten times less inhibitory on oxotremorine-induced salivary secretion than oxybutynin in rats. Further evaluation of antimuscarinic effects on bradycardia and pressor in pithed rats, and on tremor in mice, demonstrated that YM-58790 can be useful for treatment of urinary urge incontinence as a bladder-selective M3 antagonist with fewer side effects. PMID- 9734317 TI - Optimum conditions for the 13C-phenylalanine breath test. AB - We have conducted optimization studies to develop a superior 13C-phenylalanine breath test for the diagnosis of liver disease. First, we examined the optimum 13C-labeling position in phenylalanine for use in a breath test based on infrared spectroscopic detection of 13CO2 in exhaled air. L-[1-13C]Phenylalanine gave the best result. Next, a suitable dosage to give a short peak time (the time expressed in minutes at which 13CO2 excretion is maximal) after administration was determined. The 13CO2/12CO2 ratio in exhaled air after administration of 100 mg/body of L-[1-13C]phenylalanine peaked sharply at 15 min. We also examined the effect of food on the hepatic metabolism of L-[1-13C]phenylalanine. We found that a fasting period of over 7 h before the test resulted in a higher 13CO2 peak excretion. The peak appeared sooner than that in the 13C-phenacetin breath test and, therefore, the 13C-phenylalanine breath test appears preferable for the rapid evaluation of hepatic function. PMID- 9734318 TI - Kotalanol, a potent alpha-glucosidase inhibitor with thiosugar sulfonium sulfate structure, from antidiabetic ayurvedic medicine Salacia reticulata. AB - A potent natural alpha-glucosidase inhibitor called kotalanol has been isolated from an antidiabetic traditional Ayurvedic medicine, the roots and stems of Salacia reticulata Wight, through bioassay-guided separation. The structure of kotalanol was elucidated on the basis of chemical and physicochemical evidence to be the inner salt comprised of 1-deoxyheptosyl-3-sulfate anion and 1-deoxy-4-thio D-arabinofuranosyl sulfonium cation. Kotalanol was found to show more potent inhibitory activity against sucrase than salacinol and acarbose. PMID- 9734319 TI - Reduction of organochlorine contaminants from fish oil during refining. AB - Samples of crude fish oil have been refined, and the crude fish oil together with samples taken out after each step of the refining process have been analysed for organochlorine pesticides and PCB (quantified both as CB congeners and total PCB). The levels of organochlorine contaminants in fish oils remain almost constant during the neutralisation and bleaching steps of the refining process. The deodorisation step seems to cause a decrease in the amount of contaminants, especially for the most volatile compounds (alpha-HCH, lindane, HCB) where the levels were reduced to below the detection level. Concentrations of the less volatile organochlorine pesticides (dieldrin, p,p'-DDE and p,p'-DDD) and PCB are reduced to about half the concentration in the crude fish oil. PMID- 9734320 TI - Mutagenic interactions of model chemical mixtures. AB - Although current methodology for human health risk assessment assumes additive interactions among the contaminants of a complex mixture, chemical interactions may occur which produce synergistic or antagonistic effects. In this study, the mutagenic response of three f2p4l compounds, benzo(a)pyrene (B(a)P), pentachlorophenol (PCP) and 2,4,6-trinitrotoluene (TNT), were tested individually and in binary and tertiary solutions, using the Salmonella/microsome assay with each of three bacterial tester strains (TA97a, TA98, and TA100). For all strains, B(a)P was mutagenic with metabolic activation (Arochlor 1254-induced Sprague Dawley rat liver S9 fraction), TNT was mutagenic without metabolic activation, and pentachlorophenol was inactive both with and without metabolic activation. In binary and tertiary solutions, pentachlorophenol had no effect on the mutagenicity of B(a)P or TNT, independent of metabolic activation. For strain TA97a, the mutagenicity of B(a)P with metabolic activation was slightly decreased in the presence of TNT; the mutagenicity of TNT without metabolic activation was slightly decreased in the presence of B(a)P and PCP; and the mutagenicity of the tertiary solution (496 revertants/10 ug) with metabolic activation was lower than the mutagenicity of B(a)P alone (729 revertants/10 ug). The mutagenicity of B(a)P in strain TA98 with activation was inhibited by the addition of TNT. Studies conducted using several concentrations of TNT or B(a)P indicate that the inhibition of B(a)P mutagenicity was increased as the concentration of TNT increased. Assays performed using four concentrations of S9 indicated the inhibition of B(a)P mutagenicity was relatively unaffected by the level of S9. The data suggest that an interaction in the presence of TNT limits the concentration of B(a)P that is capable of reaching or binding with bacterial DNA. PMID- 9734321 TI - Concentrations of persistent lipophilic compounds in fish are determined by exchange across the gills, not through the food chain. AB - Uptake of persistent lipophilic toxicants in fish occurs via the food and by transfer across the body surface, notably the gills. Flux rates of most lipid soluble toxicants across the gills is rapid and the animal must eat at very high rates for feeding to have a significant effect on toxicant concentration in the body. The relative rates of uptake via feeding and transfer across the gills are analyzed from a theoretical and experimental standpoint. At the low feeding rates typical of fish, the uptake of toxicants in the food can be ignored when estimating toxicant body concentration. PMID- 9734322 TI - Occurrence of pesticides in Danish shallow ground water. AB - Shallow ground water from four different catchment areas, two sandy and two clayey locations, were investigated for content of pesticides and -degradation products. The samples were taken from screens in extraction wells placed from 1.5 to 5 meters below surface 46 different compounds were included in the study, and the analysis were performed with liquid chromatography mass spectrometry (LC-MS) with detection limits below 0.01 microgram/L. In total more than 300 samples were analysed and the most frequently found compounds being atrazine and its degradation products, bentazone, MCPA, metamitron, isoproturon and simazine. 23 out of the 46 compounds were detected in one or more samples. Four different sulfonylurea herbicides were included in the study as they to some extent have substituted phenoxyacid herbicides in agricultural practice and in much lower dosage. Sulfonylurea herbicides did not occur in any samples. PMID- 9734323 TI - The ecotoxicity and the biodegradability of lactic acid, alkyl lactate esters and lactate salts. AB - The ecotoxicity of lactic acid, its alkyl esters and selected metal salts was studied experimentally with the micro alga Selenastrum capricornutum, the crustacean Daphnia magna and the fish species Brachydanio rerio and Pimephales promelas. In addition, the biodegradation of lactate esters was also studied. The aim of the study was to provide predicted environmental data for additional alkyl homologues and metal salts. The ecotoxicity data are evaluated by means of Structure Activity Relations (SAR), using literature data on a non-polar narcotic mechanism of toxicity as a baseline for comparison. Lactate salts were evaluated by comparison to the toxicity of the metal ion. For the fish and D. magna, it was evident that methyl, ethyl, propyl and to a lesser extent butyl lactate were slightly more toxic in comparison to baseline non-polar narcotic toxicity data. The toxicity tests carried out with lactate-salts demonstrated clearly that the toxicity in standard tests is only determined by the associated cation and not by the lactate part. Lactic acid and its alkyl esters were degraded for more than 60% in the ready biodegradability tests and from the data presented, it is evident that the majority of alkyl lactates are readily biodegradable. The results presented in this study indicate that alkyl lactate esters show some differences in their ecotoxicity when compared to non polar narcotic compounds in but that these differences are generally small. When aquatic toxicity is considered together with their rapid tendency to biodegrade, it is concluded that lactate esters show generally favourable environmental characteristics. PMID- 9734325 TI - Correlation between whole blood cholinesterase activity and cerebral cortex cholinesterase activity in rats treated with parathion. AB - Organophosphate and carbamate insecticides are inhibitors of cholinesterases (ChE). The depression of blood ChE activity is frequently used as indicative of exposure to these chemicals. However, it is not known whether the inhibition of blood ChE activity reflects the inhibition of ChE in target tissues (e.g. brain and muscle). In this study we investigated the possibility of using whole blood ChE activity to predict frontal cerebral cortex ChE activity in rats treated with parathion. Twenty four hours after the intraperitoneal administration of several doses of parathion, the activity of ChE in whole blood and the activity of ChE in frontal cerebral cortex were determined in each animal. A high correlation between the two parameters was found (r = 0.96, p < 0.05) and the model of linear regression fitted to the data accounted for 93% of its variability. Thus, these results seem to indicate that 24 hours after the treatment with parathion the effects induced on whole blood ChE activity may be used to predict the effects caused on frontal cerebral cortex ChE activity. PMID- 9734324 TI - Effects of dissolved organic matter (DOM) on the bioconcentration of organic chemicals in aquatic organisms--a review. AB - Current knowledge on the effects of dissolved organic matter (DOM) on the bioconcentration of organic chemicals in aquatic animals (water fleas, mussels, amphipods and fish) is summarized. A graphical representation of the available data gives an overview of the magnitude of the observed effects. Most of the studies have shown decreases in bioconcentration in the presence of DOM (2 to 98% relative to DOM-free controls). However, at low DOM levels, up to 10 mg/L, also enhancements of bioconcentration due to DOM, ranging from 2 to 303% have been reported. Generally, the change in BCFW (Bioconcentration factor on a wet weight basis) per mg/L DOC was most pronounced at low levels of DOC. The data also show that DOM from different sources with different characteristics and quality can lead to substantial variations in the bioconcentration of organic compounds at comparable levels of DOC. While decreases in bioconcentration have generally been attributed to a lack of bioavailability of DOM-bound chemical, no mechanisms have been proposed to explain increased uptake of xenobiotics caused by DOM. PMID- 9734326 TI - Airway smooth muscle contractile, regulatory and cytoskeletal protein expression in health and disease. AB - The major part of research dealing with the biophysical and biochemical properties of airway smooth muscle is based on the assumption that the cells constituting the tissue are homogenous. For striated muscle this has been shown untenable. In recent years almost every property of vascular smooth muscle has been also demonstrated to be heterogeneous. This realization has been late in arriving on the airway smooth muscle research scene. Our own studies have shown that mechanical properties are, in quantitative terms, heterogeneously distributed down the airways and that contractility, for example, in extrapulmonary and intrapulmonary airways differs markedly. Another indication of heterogeneity is derived from studies of the biochemical properties of airway smooth muscle cells (ASMCs) in culture. Dramatic changes in phenotype expression were found with days in culture. Just after isolation from the tissue, the cells were of contractile type and contained mature isoforms of contractile, regulatory and cytoskeletal proteins. After the fourth day in culture the cellular phenotype changed such that contractile filaments diminished rapidly with smooth muscle isoforms being replaced by non-muscle isoforms. The cell assumed secretory or synthetic properties and commenced proliferating rapidly. It is possible that similar changes in phenotype could occur in vivo in cells undergoing hypertrophy or hyperplasia. Thus, a thickened medial layer of the type seen in the walls of airways from asthmatic airways is not necessarily one endowed with increased contractility and, in fact, the latter may be subnormal. Finally, using the so called motility assay, we studied the velocity of translation of actin filaments by myosin molecules obtained from antigen-sensitized and control airway smooth muscle. We found no change in maximum velocity of actin translation. This was under conditions where the myosin light chain (MLC) was fully phosphorylated. However, in these tissues we found heterogeneity in myosin light chain kinase (MLCK) content which, we inferred, accounted for the difference in shortening velocity between control and sensitized muscle strips in vitro. PMID- 9734327 TI - Myosin heavy chain isoforms and dynamic contractile properties: skeletal versus smooth muscle. AB - Myosin, one of the primary contractile muscle proteins, displays molecular, enzymatic, structural, functional and regulatory variability. This variability has been shown to account for a significant amount of the functional uniqueness of skeletal and smooth muscle. However, the universal generation of force and/or shortening by these two muscle types belies the ever-increasing number of known distinct differences that bring this about. Thus, the notion that the functional roles of skeletal and smooth muscle, their development and regulation, all appear to be uniquely applicable for their physiological purpose no longer appears heretical. This manuscript presents a cursory overview of the numerous ways in which these two types of muscle use a host of myosin molecules to bring about a common result, force generation and/or shortening. PMID- 9734328 TI - Cross-bridge cycling kinetics, actomyosin ATPase activity and myosin heavy chain isoforms in skeletal and smooth respiratory muscles. PMID- 9734329 TI - The molecular mechanics of smooth muscle myosin. AB - Smooth muscle cells are capable of generating forces comparable to those of skeletal muscle cells but with far less myosin, the molecular motor that powers muscle contraction. This unique capability may be inherent to the myosin molecule. We have directly characterized the molecular mechanics of smooth muscle myosin using new technologies developed to measure the forces generated by these proteins. The data help explain the differences in force and velocity in whole smooth and skeletal muscles. PMID- 9734330 TI - Myosin heavy chain transitions during development. Functional implications for the respiratory musculature. AB - The myosin heavy chain (MHC) exists as multiple isoforms that are encoded for by a family of genes. The respiratory musculature demonstrates muscle-specific and temporally-dependent changes in MHC isoform expression during maturation. Developmental expression of MHC isoforms correlate well with postnatal changes in actomyosin ATPase activity, specific force generation (P0/CSA), maximum unloaded velocity of shortening (V0) and and fatigue resistance. More specifically, as the expression of MHCneonatal declines and MHC2A, MHC2X, and MHC2B increase, actomyosin ATPase activity, P0/CSA, V0, and muscle fatigability increase. The increase in actomyosin ATPase activity with maturation is partially offset by a postnatal increase in oxidative capacity; however, as fatigue resistance declines with development it is apparent that the energy costs of contraction are not fully matched by an increase in energy production. Developmental transitions in smooth muscle MHC phenotype also occur although their functional importance remains unclear. PMID- 9734332 TI - Cloning and sequencing of three C-type lectins from body surface mucus of the land slug, Incilaria fruhstorferi. AB - Three C-type lectins of 15 kDa were isolated from the water-soluble fraction (WSF) of the body surface mucus of the land slug, Incilaria fruhstorferi. Based on their partial amino acid sequences, the nucleotide sequences of cDNAs encoding these lectins, named incilarin A, B and C, were determined. cDNAs of incilarin A, B and C consisted of 673, 663 and 715 bp, and deduced amino acids were 150, 149 and 156 residues, respectively. All three lectins had signal peptides of 17 amino acid residues at their N-termini. They showed 44-55% amino acid sequence identity with each other, and lower but significant homology with the other animal C-type lectins and antifreeze protein. Incilarin A and B seem to possess two intramolecular disulfide bonds in the carbohydrate-binding domain (CRD) conserved among the animal C-type lectins, however, one of these bonds is absent in incilarin C. PMID- 9734333 TI - Porcine kidney betaine aldehyde dehydrogenase: purification and properties. AB - Significant betaine aldehyde dehydrogenase activity was found in porcine kidney. The enzyme was purified 320-fold with an overall recovery of 11%. It had a specific activity of 115.8 nkats/mg protein and proved to be homogeneous by SDS PAGE with a subunit molecular mass of 52 kDa. IEF studies showed three bands with pI values of 5.74, 5.68 and 5.58, respectively. The enzyme was stable in a pH range between 5.0 and 10.0 and the optimum pH was 9.5. The reaction is highly specific for NAD+ and betaine aldehyde, although acetaldehyde, butyraldehyde and glyceraldehyde can be used. Estimated values of Km at pH 8.0 and 25 degrees C were 127 microM for betaine aldehyde and 40 microM for NAD+. The reaction could not be reversed even at high glycine betaine concentrations. The enzyme was not activated by salts at high concentrations but it was salt tolerant-retaining 50% of maximal activity at 1.0 M K+ and Na+. It is inferred that salt tolerance is an essential property for an enzyme participating in the cellular synthesis of an osmoprotectant. Proline, glycerol, sucrose and mannitol had a little effect on the enzyme activity while glycine betaine had an inhibitory effect. PMID- 9734331 TI - Atypical phosphorylcholine-reactive protein from Atlantic salmon, Salmo salar L. AB - A phosphorylcholine-reactive protein was isolated from serum of Atlantic salmon (Salmo salar L.) by affinity chromatography on a phosphorylcholine-conjugated Sepharose column followed by elution with phosphorylcholine. Based on the method used we describe the isolated protein as salmon phosphorylcholine-reactive protein (salmon PRP). Salmon PRP has calcium-independent binding to phosphorylcholine. The protein exists in a monomeric and dimeric form with molecular weight of approximately 80 and 160 kD, respectively. Separation of the protein preparation on SDS-PAGE under reducing conditions resulted in disappearance of the 80 and 160 kD bands and appearance of a major protein band of approximately 100 kD. The N-terminal amino acid sequences of the non-reduced 80 and 160 kD bands and the reduced 100 kD band were identical. Apart from the dimeric form, the molecular weight of salmon PRP and its appearance on SDS-PAGE is similar to human plasminogen. Comparison of the sequence in a protein database resulted in approximately 50% identity with human and bovine plasminogen. In addition, cross-reactivity between antibodies to human plasminogen and salmon PRP was demonstrated. Thus, salmon PRP appears to be different from other phosphorylcholine-reactive proteins which are mostly reported to be CRP-like proteins with calcium-dependent binding to phosphorylcholine, pentameric ring structure and sequence homology between species. Whether salmon PRP is a new type of phosphorylcholine-binding protein with an unknown function or a plasminogen like protein with binding specificity for phosphorylcholine calls for further investigation. PMID- 9734334 TI - Glyceraldehyde-3-phosphate dehydrogenase from Tetrahymena pyriformis: enzyme purification and characterization of a gapC gene with primitive eukaryotic features. AB - Glyceraldehyde-3-phosphate dehydrogenase (GAPDH, EC.1.2.1.12) was purified to electrophoretic homogeneity from an amicronucleated strain of the ciliate Tetrahymena pyriformis using a three-step procedure. The native enzyme is an homotetramer of 145 kDa exhibiting absolute specificity for NAD. In its catalytic properties it is similar to other glycolytic GAPDHs. Chromatofocusing analysis showed the presence of only one basic GAPDH isoform with an isoelectric point of 8.8. Western blots using a monospecific polyclonal antibody raised against the T. pyriformis GAPDH showed a single 36-kDa band corresponding to the enzyme subunit in the cytosolic protein fraction of this strain and the closely related species, both from the class Oligohymenophorea, Paramecium tetraurelia. No bands were immunodetected in the ciliate Colpoda inflata (class Colpodea) and in the diverse eukaryotes and eubacteria tested. A 0.5-kb DNA fragment which corresponds to an internal region of a gapC gene was generated by polymerase chain reaction using cDNA of T. pyriformis as template. This gene codes for a basic GAPDH protein with eukaryotic-diplomonad signatures and exhibits a codon usage biased in the manner typical for T. pyriformis genes. Southern blots performed both under homologous and heterologous conditions using this amplified cDNA fragment as a probe, indicated that it should be the only gapC gene present in the macronuclear genome of this ciliate, its expression being confirmed by Northern blot analysis. These results are discussed in connection with the peculiar genomic organization of ciliates and in the context of protist evolution. PMID- 9734335 TI - Isolation and characterization of cDNA encoding chicken egg yolk aminopeptidase Ey. AB - Aminopeptidase Ey (EC 3.4.11.20) from chicken (Gallus gallus domesticus) egg yolk is a homodimeric exopeptidase with a broad specificity for N-terminal amino acid residues at P1 position of the substrate. Aminopeptidase Ey is a 300-k metalloexopeptidase, containing 1.0 g atom of zinc per mole of a subunit with a relative molecular mass of 150 k. A full-length cDNA was cloned from chicken (female) liver cDNA library. Analysis of the 3196-base pairs (bp) nucleotide sequence of the cDNA revealed a single open reading frame coding for 967 amino acid residues. The coding region of aminopeptidase Ey gene, apdE, occupies 2901 bp of the cDNA. The predicted amino acid sequence of the enzyme is 66, 65, 64 and 63% identical with those of aminopeptidases N (EC 3.4.11.2) from human, pig, rabbit and rat, respectively. Aminopeptidase Ey contains the metallo-binding sequence motif, His-Glu-Xaa-His, found in zinc metallopeptidases. Zinc binding sites, His-386, His-390 and Glu-409, and catalytic site, Glu-387, were conserved in the homologous aminopeptidases N. PMID- 9734336 TI - Compartmentation and kinetics of urea cycle enzymes in porcine enterocytes. AB - We have recently reported the synthesis of urea from ammonia, glutamine and arginine in enterocytes of postweaning pigs. The present study was conducted to determine the compartmentation and kinetics of urea cycle enzymes in these cells. Carbamoyl phosphate synthase I (CPS I) and ornithine carbamoyltransferase (OCT) were located exclusively in mitochondria, whereas argininosuccinate synthase (ASS) and argininosuccinate lyase (ASL) were found in the cytosol. Arginase isozymes were present in both the cytosol and mitochondria of enterocytes, and differed in their sensitivity to heat inactivation. Except for OCT, Vmax values of urea cycle enzymes were much lower in enterocytes than in the liver of pigs, and vice versa for their Km values. Because of a low rate of ureagenesis in enterocytes compared with the liver, intestinal urea cycle enzymes may function primarily to synthesize citrulline. The co-localization of CPS I and OCT and a high activity of OCT in enterocyte mitochondria favors the intestinal synthesis of citrulline from ammonia, HCO3- and ornithine. Low activities of cytosolic ASS and ASL minimize the conversion of citrulline into arginine and therefore, the recycling of citrulline into ornithine via arginase in postweaning-pig enterocytes. These kinetic properties of intestinal urea cycle enzymes maximize the net synthesis of citrulline from glutamine and explain the release of large amounts of citrulline by the pig small intestine. The two compartmentally separated arginase isozymes in enterocytes may play an important role in regulating the intestinal metabolism of proline, nitric oxide and polyamines. PMID- 9734337 TI - New pathway of heparan sulphate degradation. Iduronate sulphatase and N sulphoglucosamine 6-sulphatase act on the polymer chain prior to depolymerisation by a N-sulpho-glucosaminidase and glycuronidases in the mollusc Tagelus gibbus. AB - It has previously been shown that in the mollusc Anomalocardia brasiliana the desulphation of chondroitin sulphate precedes its depolymerisation by beta glucuronidase and beta-N-acetylgalactosaminidase (Sousa Jr. et al. J. Biol. Chem. 1990;265:20150-20155). This led us to investigate whether in molluscs, sulphatases also act on heparan sulphate before its depolymerisation by glycosidases. Radioactively labelled [35S]heparan sulphate was extensively degraded by enzyme extracts prepared from the mollusc Tagelus gibbus. Several enzymes acting in concert degrade the compound to inorganic sulphate, glucosamine N-sulphate, N-acetylglucosamine-6 sulphate and other oligosaccharide products. These results indicate the presence of iduronate sulphatase, N-sulphoglucosamine 6-sulphatase alpha-N-sulphoglucosaminidase, beta-glucuronidase and alpha-L iduronidase. The di- and mono-saccharide composition of the oligosaccharides were analysed with the aid of heparitinase II from Flavobacterium heparinum. These analyses led to the characterisation of two sulphatases that act on the polymer chain removing sulphates from the C-2 position of iduronic acid residues and the C-6 position of the glucosamine moieties, respectively. The different enzymes were partially fractionated by ion exchange chromatography and molecular sieving. These results led to the proposition of a new pathway of degradation of heparan sulphate where sulphatases act directly on the polymer chain which is then depolymerised by several glycosidases. PMID- 9734338 TI - Fatty acid binding protein, a major protein in the flight muscle of migrating western sandpipers. AB - Migratory flight in birds is fueled primarily by fatty acid oxidation imposing a requirement for high rates of fatty acid: (a) transport; (b) uptake; and (c) delivery to intracellular sites of beta-oxidation. Muscle fatty acid binding protein (M-FABP) is a cytosolic protein involved in the intracellular transport of fatty acids. Its expression appears to be correlated with muscle fatty acid oxidation capacity. The M-FABP was isolated for the first time from a long distance migrant bird using: (i) size exclusion; (ii) anion exchange; and (iii) hydroxyapatite chromatography. M-FABP has a molecular weight of approximately 14,000 Da and an isoelectric point of pH 4.8. A partial amino acid sequence of the protein demonstrated homology to M-FABPs from other species (80% identical to human heart FABP). It was estimated that M-FABP comprises approximately 14 and 21% of total cytosolic protein of the pectoralis and heart, respectively; the highest values yet reported from any vertebrate muscle. The abundance of M-FABP in these tissues suggests that the protein may play a key role in fatty acid supply during endurance flight. Thus, it is proposed that a seasonal increase in M-FABP expression could be a component of physiological preparation for migration. PMID- 9734339 TI - Comparative tyrosine degradation in Vibrio cholerae strains. The strain ATCC 14035 as a prokaryotic melanogenic model of homogentisate-releasing cell. AB - The relationship between L-tyrosine catabolism and melanin formation was studied in the Vibrio cholerae strains ATCC 14035 and CECT 557. It is shown that both strains degrade L-tyrosine by the same pathway as eukaryotic cells, giving homogentisate as intermediate. ATCC 14035, an O1 strain, which is not able to grow using L-tyrosine as sole carbon and energy source, but it forms pyomelanin from homogentisate. The second strain, which is non-O1, is able to grow using L tyrosine as sole carbon and energy source, but it does not form any pigment. Both strains contain all the enzymes involved in the L-tyrosine catabolism. The three late enzymes of the pathway, homogentisate oxygenase, maleylacetoacetate isomerase and fumarylacetoacetate hydrolase, are induced by L-tyrosine, but the degree of induction is much lower in the ATCC 14035 strain. Thus, the distal part of the pathway becomes the rate-limiting steps in the L-tyrosine catabolism, explaining homogentisate accumulation and pyomelanogenesis in this strain. It is proposed that V. cholerae might be a useful prokaryotic model to show that alkaptonuria and other diseases related to L-tyrosine metabolism could occur in animals even when no particular enzyme involved in that pathway is lacking. PMID- 9734340 TI - Fatty acid composition and cholesterol concentration in tissues of white-tailed deer (Odocoileus virginianus) as influenced by lactation, age, and season of the year. AB - The purpose of this study was to determine effects of lactation, season, and age on fatty acid compositions of adipose tissue (subcutaneous and perirenal), liver, and muscle (m. longissimus dorsi), and on cholesterol concentration of liver and muscle, of female white-tailed deer (Odocoileus virginianus). Lactation did not affect fatty acid composition in adipose tissue or muscle, but in liver, weight percentages of 18:2 were lower, and of 20:4 higher in non-lactating does. Increased age (fawns, yearlings, 2 and 3+(-)years old) decreased 14:0, 15:0, and 16:0 in subcutaneous adipose tissue; decreased 14:0, 15:0, 16:1, 18:2, and increased 18:0 in perirenal adipose tissue; increased 18:1 and decreased 18:2 in liver; and increased 18:1, 18:3, and 20:4, and decreased 18:2 in muscle. Season of the year had little effect on adipose tissue and muscle fatty acids. Liver of fall season does had greater concentrations of most fatty acids than winter does, and cholesterol concentration was greatest in liver of winter does. It was concluded that season and lactation minimally affect fatty acid composition in adipose and muscle, whereas changes in saturated and unsaturated fatty acids occur with increased age. Also, liver fatty acids are influenced by each parameter studied, but cholesterol varies only with season. PMID- 9734341 TI - Purification and properties of embryonic cysteine proteinase which participates in yolk-lysis of Xenopus laevis. AB - The study reported here aimed to purify a cysteine proteinase from neurula embryos of Xenopus laevis, since this enzyme was thought to be involved in yolk lysis in developing embryos. The purification procedure consisted of fractionation of an embryonic extract by means of 30-90% ammonium sulfate, chromatography on diethylaminoethyl cellulose and carboxymethyl cellulose, gel filtration on Sephadex G-75 and affinity chromatography on concanavalin A agarose. The purified enzyme had a molecular weight of 30 kDa according to both SDS-PAGE and Sephadex G-75 gel-filtration and an optimum pH of 5.5, and it preferentially cleaved the synthetic substrate, Z-Phe-Arg-MCA. Its activity was inhibited by Z-Phe-Phe-CHN2, a specific cathepsin L inhibitor, as well as by leupeptin and E-64. The NH2-terminal amino acid sequence of the enzyme was similar to that of chicken cathepsin B. These characteristics indicate that the purified enzyme is a member of the cysteine proteinase family. The antibody raised against the purified enzyme specifically stained a 30 kDa protein of neurula embryo extracts on immunoblot tests. The enzyme effectively digested Xenopus yolk proteins when the NaCl concentration in test solutions was 0.2 M. It was also confirmed that cysteine proteinase inhibitors inhibited yolk-lysis by the enzyme. PMID- 9734343 TI - Purification and characterization of Microcystis aeruginosa (freshwater cyanobacterium) lectin. AB - Microcystis aeruginosa, strain M228, a laboratory culture of freshwater cyanobacterium, showed hemagglutinating activity against rabbit, horse and human ABO erthrocytes. Crossed absorption tests revealed the presence of a single type of lectin in the extract of M228 strain cells. The lectin, termed MAL, was purified in combination with the affinity chromatography on acid-treated agarose gel and the gel permeation chromatography in an electrophoretically pure form. MAL was a glycoprotein containing 7.8% neutral sugars and was composed of a single polypeptide having a molecular weight of 57 kDa. Isoelectric point was estimated to be pH 6.4. Hemagglutinating activity of the lectin was inhibited effectively by N-acetyl-D-galactosamine and by glycoproteins. D-galactose and lactose also showed moderate inhibitory activity. The destruction of the hemagglutinating activity by a 2-mercaptoethanol treatment suggests the presence of intra-chain disulfide bond(s) essential for the activity in the molecule. The sequence of the amino-terminal region of MAL was determined as Val-Leu-Ala-Ser Leu-Val-Ser-Thr-Ser-Gln-Ala-Gly-Ser-Leu-Glu-Leu-Leu- Ala [corrected]. PMID- 9734342 TI - Primary structure and characterization of a bullfrog visual pigment contained in small single cones. AB - A cDNA fragment encoding a putative visual pigment (FCV pigment) was isolated from the bullfrog, Rana catesbeiana. Its deduced amino acid sequence shows high similarities to those of short wavelength-sensitive pigments such as human blue-, chicken violet- and goldfish ultraviolet-sensitive pigments. An antiserum against its C-terminal amino acid sequence recognized the outer segments of small cone photoreceptor cells without oil droplets. It is suggested that the FCV pigment is a short wavelength-sensitive pigment contained in small single cones which have not been characterized previously. PMID- 9734344 TI - Peroxidase activity in Malpighian tubules of Triatoma infestans Klug. AB - Benzidine and diamino benzidine (DAB) oxidation, typically performed by peroxidases, was demonstrated by light and electron microscopy in peroxisomes, mitochondria and membranous structures which occurred in close contact with urate crystals in Malpighian tubules of nymphs and adults of Triatoma infestans. Peroxisomes were predominantly identified in cells of the distal region of the tubules, which is engaged in excretory mechanisms. DAB oxidation in mitochondria, even in the absence of hydrogen peroxide, may indicate the existence of a mitochondrial peroxidase and possibly a cytochrome c peroxidase. The localization of the extracellular membranous structures appeared restricted to the lumen of the proximal region of the tubules and they were assumed to be remnants of endoplasmic reticulum containing peroxidases. PMID- 9734345 TI - Antidotal effect of grape juice (Vitis vinifera) on ochratoxin A caused hepatorenal carcinogenesis in mice (Mus musculus). AB - Oral administration of ochratoxin A to young weanling mice (Mus musculus) caused several haematological changes and induced hepatoma and renal carcinoma. Concurrent administration of berry and leaf juice of the common grape (Vitis vinifera) to mice together with ochratoxin A significantly reduced the hepatic and renal damage caused by ingestion of this mycotoxin. None of the animals receiving berry/leaf juice of V. vinifera showed the formation of hepatorenal carcinoma whereas 25% of animals receiving only ochratoxin A developed well differentiated renal carcinoma and hepatic lesions. PMID- 9734346 TI - Why do only some of the young adults with bronchial hyperreactivity wheeze? AB - The significance of nonspecific bronchial hyperreactivity (BHR) is a controversial issue in asthma. The natural history of BHR has not been investigated adequately although its importance as a cross-sectional risk factor for asthma is widely accepted. This paper investigates the risk factors for wheeze among people with BHR. Subjects were young adults who had participated in the second phase of the European Community Respiratory Health Survey in Melbourne, Australia. We compared the participants with wheeze and BHR (n=186) to those with asymptomatic BHR (n=66). Information was collected on sociodemographic factors, family history of asthma, and relevant environmental factors using an interviewer-administered questionnaire. Atopy to a range of aeroallergens was examined by skin prick tests. Risk factors were examined by adjusting the odds ratios (OR) by a logistic regression to control for confounding effects. Parental asthma (OR=4.2), keeping pets during childhood (OR=3.3), allergy to house dust mite (OR=2.7), allergic rhinitis (OR=2.6), and having ever smoked (OR=2.4) were associated with an increased risk of wheeze, independent of the other factors examined. When allergic rhinitis was not included as an explanatory variable, being atopic to any of the allergens assessed was found to increase the risk of current wheeze (OR=4.8). Allergic rhinitis may represent an intermediate stage in the natural history of BHR. Avoidance of pets during childhood, not smoking, and taking steps to minimize dust exposure are likely to prevent the progression from asymptomatic BHR to asthma. PMID- 9734347 TI - Downregulation of the expression of intercellular adhesion molecule (ICAM)-1 on bronchial epithelial cells by fenoterol, a beta2-adrenoceptor agonist. AB - Inflammatory airway disorders, such as asthma and chronic bronchitis, are characterized by overexpression of adhesion molecules on airway epithelial and endothelial cells. This phenomenon is associated with increased adherence and activation of polymorphonuclear leukocytes (PMNs). With the knowledge that beta2 adrenoceptor agonists demonstrate some anti-inflammatory activity in vitro, the present study was designed to evaluate whether fenoterol could interfere with adhesion molecule expression on airway epithelium. Human bronchial epithelial cells (HBECs), obtained by protease digestion from surgically resected bronchi, were stimulated with human recombinant interferon-gamma (rh IFN-gamma) in the presence of (a) fenoterol (10(-12)-10(-5) M); (b) dexamethasone (10(-12)-10(-5) M); and (c) fenoterol and dexamethasone. Because desensitization after high-dose exposure to agonists has been described for many membrane-associated receptors, in additional sets of experiments HBECs were preexposed to fenoterol and, as control, to dexamethasone for 8 hr, then washed and stimulated with rh IFN-gamma in the presence of fresh drugs. The cells were harvested after 24-hr culture and stained by specific monoclonal antibodies. The intensity of intercellular adhesion molecule-1 (ICAM-1) expression was then measured by flow cytometry analysis and expressed as mean fluorescence channel (mfc). The significant increase in ICAM-1 expression on HBECs induced by rh IFN-gamma was inhibited, in a dose-dependent manner, by the two drugs, but fenoterol was more efficient than dexamethasone at all of the concentrations tested (p < 0.05, all comparisons). In addition, the inhibitory activity of fenoterol was not enhanced by the simultaneous presence of dexamethasone in rh IFN-gamma-stimulated HBEC cultures (p > 0.05, all comparisons). Finally, preexposure to fenoterol or to dexamethasone did not induce any modification of the inhibitory effect of the two drugs on ICAM-1 expression (p > 0.05, all comparisons). These results suggest that clinical efficacy of fenoterol in patients with obstructive lung disease may include downregulation of adhesion molecule expression on airway epithelial cells. PMID- 9734348 TI - Psychological and family characteristics of adolescents with vocal cord dysfunction. AB - Vocal cord dysfunction (VCD) is a respiratory disorder often confused with asthma. Although previous case reports have implicated family and/or individual psychopathology in the etiology of this condition, this is the first paper to present prospective, case-control empirical data on a group of pediatric patients with VCD. A case-control methodology was employed to examine patients in terms of both family and individual functioning. Results indicate that patients with VCD were not different from asthmatic controls on measures of family functioning. However, they did experience significantly higher levels of anxiety and received a higher number of anxiety-related diagnoses such as separation anxiety and generalized anxiety disorder on a structured psychiatric interview. The nature of the relation between VCD and psychological symptoms in children is discussed. Etiologic and treatment issues are examined in the context of the findings. PMID- 9734350 TI - Reasons for delay in seeking treatment for acute asthma: the patient's perspective. AB - Increased morbidity and mortality due to asthma suggested the need to investigate whether persons with asthma report delay in seeking medical care during acute asthma exacerbations and the reasons they gave for delay. We interviewed 95 asthmatic adults, 36 men and 59 women, using a critical incident technique to discover how patients responded to acute asthma symptom episodes. Subjects were interviewed once per month for a total of three interviews. All subjects had physician-diagnosed asthma for a mean of 16.8+/-14.5 years. Eighty-six percent of the sample (n=82) reported delay in seeking medical care for severe asthma symptoms. Seven reasons for delay were identified: uncertainty, disruption, minimization, fear of systemic corticosteroid, previous bad experiences in emergency departments, the need to "tough it out" alone, and economic reasons. Seventy-one subjects (86.5%) reported three or more reasons for delay. Thirteen subjects (16%) identified pivotal episodes in which they realized they could die from asthma and as a result, no longer delayed. People with asthma often delay seeking urgent care for acute episodes for a variety of reasons. Some of these reasons are modifiable. Clear directions from health professionals to guide patients in responding to acute asthma episodes are needed. Asthma action plans written by the primary physician may be a positive agent of change for those who delay. PMID- 9734349 TI - The risk for outpatient antibiotic-treated infections following a course of oral corticosteroids among children with asthma. AB - Short courses of oral corticosteroids are widely used to treat asthma. The objective of this study was to assess if one course of oral corticosteroids increases asthmatic children's risk for infections treated with outpatient antibiotics. Using New York State Medicaid claims data on asthmatic children 2-15 years old, we made cohorts of oral corticosteroid users and nonusers. We determined the percentage of children who filled antibiotic prescriptions in the 30 days after index dates. Index dates were dates oral steroids were started (for steroid users) or matched dates (for nonusers). Odds ratios were adjusted for age, month of index date, and prior antibiotic use. Among children not receiving antibiotics on index dates, antibiotic prescriptions were filled in the next 30 days for 438 (20%) of 2145 steroid nonusers and 130 (19%) of 698 steroid users (p=0.30); compared to nonusers, steroid users had an adjusted odds ratio of subsequent antibiotic use of 0.92 (95% confidence interval [CI] 0.73-1.15). Among children receiving antibiotics on index dates, antibiotic prescriptions were filled in the next 30 days for 116 (26%) of 451 steroid nonusers and 50 (19%) of 260 steroid users (p=0.05); compared to nonusers, steroid users had an adjusted odds ratio of subsequent antibiotic use of 0.65 (95% CI 0.53-0.97). We conclude that one course of oral corticosteroids does not increase asthmatic children's risk for infections treated with outpatient antibiotics. PMID- 9734351 TI - Burden of wheezing illness among U.S. children reported by parents not to have asthma. AB - We examined the 12-month prevalence of asthma and wheezing among U.S. children and compared the illness-related burden of children who wheezed with and without an asthma diagnosis. Data were obtained in a cross-sectional telephone survey that tested the performance of a health interview designed to identify children with chronic health conditions. Respondents were 712 primary caretakers of 1388 children under 18 years old in a national probability sample selected by random digit dialing. Although 51 children identified with asthma and wheezing had more episodes, sleep disturbances, and attacks that limited speech, and received more medical treatment for wheezing than 69 children with wheezing alone, the "undiagnosed" children appeared to be only somewhat less affected by their wheezing. Repeat episodes and burden experienced by children with wheezing alone suggest that the asthma prevalence may be underestimated if based only on the diagnostic label. PMID- 9734352 TI - Direct evidence that LTC4 and LTB4 but not TXA2 are involved in asthma attacks in children. AB - There are substantial numbers of reports showing that leukotrienes (LTs) play important roles in adult asthma. No definite evidence has been demonstrated that LTs are involved in asthma attacks in children, although it is highly expected. In this report, we demonstrated that the levels of LTB4 and LTC4 but not thromboxane B2 (TXB2), a stable metabolite of TXA2, were significantly elevated in the bronchoalveolar lavage fluid, which was obtained from intubated and mechanically ventilated children with severe asthma attacks. This is direct evidence that LTB4 and LTC4 predominantly participate in asthma attacks in pediatric patients. PMID- 9734353 TI - Direct binding of CDC20 protein family members activates the anaphase-promoting complex in mitosis and G1. AB - Activation of the anaphase-promoting complex (APC) is required for anaphase initiation and for exit from mitosis. We show that APC is activated during mitosis and G1 by two regulatory factors, hCDC20 and hCDH1. These proteins directly bind to APC and activate its cyclin ubiquitination activity. hCDC20 confers a strict destruction-box (D-box) dependence on APC, while hCDH1 shows a much more relaxed specificity for the D-box. In HeLa cells, the protein levels of hCDC20 as well as its binding to APC peak in mitosis and decrease drastically at early G1. Thus, hCDC20 is the mitotic activator of APC and directs the degradation of substrates containing the D-box. The hCDH1 protein level remains constant during the cell cycle and may target specific substrates lacking the D box in G1, such as polo-like kinase, for ubiquitination. PMID- 9734354 TI - CLB5-dependent activation of late replication origins in S. cerevisiae. AB - Replication origins in chromosomes are activated at specific times during the S phase. We show that the B-type cyclins are required for proper execution of this temporal program. clb5 cells activate early origins but not late origins, explaining the previously described long clb5 S phase. Origin firing appears normal in cIb6 mutants. In clb5 clb6 double mutant cells, the late origin firing defect is suppressed, accounting for the normal duration of the phase despite its delayed onset. Therefore, Clb5p promotes the timely activation of early and late origins, but Clb6p can activate only early origins. In clb5 clb6 mutants, the other B-type cyclins (Clb1-4p) promote an S phase during which both early and late replication origins fire. PMID- 9734356 TI - The mutagenesis proteins UmuD' and UmuC prevent lethal frameshifts while increasing base substitution mutations. AB - Error-prone DNA repair consists of replicative filling-in of DNA gaps carrying lesions. We have reconstituted E. coli SOS error-prone repair using purified DNA polymerase III holoenzyme, SSB, RecA, UmuD', a UmuC fusion protein, and a gap lesion plasmid. In the absence of UmuDC, or without SOS induction, replication skips over the lesion, forming mostly one-nucleotide deletions. These cause translational frameshifts that usually inactivate genes. UmuD' and UmuC, in the presence of RecA and SSB, stimulate translesion replication and change its mutagenic specificity such that deletions are prevented and base substitutions are increased. This results in mutagenic but nondetrimental gap repair and provides an effective mechanism for generating genetic variation in bacteria adapting to environmental stress. PMID- 9734355 TI - MEC1-dependent phosphorylation of Rad9p in response to DNA damage. AB - In budding yeast, DNA damage can activate a checkpoint surveillance system controlled by the RAD9, RAD53, and MEC1 genes, resulting in a delay in cell cycle progression. Here, I report that DNA damage induces rapid and extensive phosphorylation of Rad9p in a manner that correlates directly with checkpoint activation. This response is dependent on MEC1, which encodes a member of the evolutionarily conserved ATM family of protein kinases, and on gene products of the RAD24 epistasis group, which have been implicated in the recognition and processing of DNA lesions. Since the phosphorylated form of Rad9p appears capable of interacting stably with Rad53p in vivo, this phosphorylation response likely controls checkpoint signaling by Rad9p. PMID- 9734357 TI - Structures of SAP-1 bound to DNA targets from the E74 and c-fos promoters: insights into DNA sequence discrimination by Ets proteins. AB - SAP-1 is a member of the Ets transcription factors and cooperates with SRF protein to activate transcription of the c-fos protooncogene. The crystal structures of the conserved ETS domain of SAP-1 bound to DNA sequences from the E74 and c-fos promoters reveal that a set of conserved residues contact a GGA core DNA sequence. Discrimination for sequences outside this core is mediated by DNA contacts from conserved and nonconserved protein residues and sequence dependent DNA structural properties characteristic of A-form DNA structure. Comparison with the related PU.1/DNA and GABPalpha/beta/DNA complexes provides general insights into DNA discrimination between Ets proteins. Modeling studies of a SAP-1/SRF/DNA complex suggest that SRF may modulate SAP-1 binding to DNA by interacting with its ETS domain. PMID- 9734358 TI - NAT, a human complex containing Srb polypeptides that functions as a negative regulator of activated transcription. AB - A complex that represses activated transcription and contains the human homologs of the yeast Srb7, Srb10, Srb11, Rgr1, and Med6 proteins was isolated. The complex is devoid of the Srb polypeptides previously shown to be components of the yeast Mediator complex that functions in transcriptional activation. The complex phosphorylates the CTD of RNA polymerase II (RNAPII) at residues other than those phosphorylated by the kinase of TFIIH. Moreover, the complex specifically interacts with RNAPII. The interaction is not mediated by the CTD of RNAPII, but is precluded by phosphorylation of the CTD. Our results indicate that the complex is a subcomplex of the human RNAPII holoenzyme. We suggest that the RNAPII holoenzyme is a transcriptional control panel, integrating and responding to specific signals to activate or repress transcription. PMID- 9734359 TI - Xeroderma pigmentosum group C protein complex is the initiator of global genome nucleotide excision repair. AB - The XPC-HR23B complex is specifically involved in global genome but not transcription-coupled nucleotide excision repair (NER). Its function is unknown. Using a novel DNA damage recognition-competition assay, we identified XPC-HR23B as the earliest damage detector to initiate NER: it acts before the known damage binding protein XPA. Coimmunoprecipitation and DNase I footprinting show that XPC HR23B binds to a variety of NER lesions. These results resolve the function of XPC-HR23B, define the first NER stages, and suggest a two-step mechanism of damage recognition involving damage detection by XPC-HR23B followed by damage verification by XPA. This provides a plausible explanation for the extreme damage specificity exhibited by global genome repair. In analogy, in the transcription coupled NER subpathway, RNA polymerase II may take the role of XPC. After this subpathway-specific initial lesion detection, XPA may function as a common damage verifier and adaptor to the core of the NER apparatus. PMID- 9734360 TI - SUMO-1 modification of IkappaBalpha inhibits NF-kappaB activation. AB - Activation of NF-kappaB is achieved by ubiquitination and proteasome-mediated degradation of IkappaBalpha. We have detected modified IkappaBalpha, conjugated to the small ubiquitin-like protein SUMO-1, which is resistant to signal-induced degradation. In the presence of an E1 SUMO-1-activating enzyme, Ubch9 conjugated SUMO-1 to IkappaBalpha primarily on K21, which is also utilized for ubiquitin modification. Thus, SUMO-1-modified IkappaBalpha cannot be ubiquitinated and is resistant to proteasome-mediated degradation. As a result, overexpression of SUMO 1 inhibits signal-induced activation of NF-kappaB-dependent transcription. Unlike ubiquitin modification, which requires phosphorylation of S32 and S36, SUMO-1 modification of IkappaBalpha is inhibited by phosphorylation. Thus, while ubiquitination targets proteins for rapid degradation, SUMO-1 modification acts antagonistically to generate proteins resistant to degradation. PMID- 9734361 TI - Recognition specificity for the bacterial avirulence protein AvrPto is determined by Thr-204 in the activation loop of the tomato Pto kinase. AB - The Pto kinase confers resistance in tomato to P. syringae pv. tomato strains expressing the AvrPto protein. Physical interaction of the Pto kinase and AvrPto protein in the plant cell initiates host defense responses. The recognition event between these two proteins is very specific; AvrPto does not interact with other closely related kinases, including the Fen kinase, which shares 80% amino acid identity with Pto. By using Pto-Fen chimeric proteins and site-directed mutagenesis, we found that Thr-204 is required for Pto interaction with AvrPto in a yeast two-hybrid system and for recognition specificity in a tobacco leaf transient assay. Substitution of Thr-204 into the Fen kinase allowed that kinase to interact with AvrPto and to confer an AvrPto-specific defense response in tobacco leaves. Thus, simple mutations appear capable of giving rise to new resistance gene specificities. PMID- 9734362 TI - Somatic mutation in individual liver cysts supports a two-hit model of cystogenesis in autosomal dominant polycystic kidney disease. AB - Autosomal dominant polycystic kidney disease (ADPKD), Type I is a common genetic disorder and an important cause of renal failure. The disease is characterized by progressive cyst formation in a variety of organs including the kidney, liver and pancreas. We have previously shown that in the case of PKD1, renal cyst development is likely to require somatic inactivation of the normal allele coupled to a germline PKD1 mutation. In this report, we have used unique reagents to show that intragenic, somatic mutations are common in hepatic cysts. All pathogenic mutations were shown to have altered the previously normal copy of the gene. These data extend the "two-hit" model of cystogenesis to include a second focal manifestation of the disease. PMID- 9734363 TI - MyD88 is an adaptor protein in the hToll/IL-1 receptor family signaling pathways. AB - The Toll-mediated signaling cascade using the NF-kappaB pathway has been shown to be essential for immune responses in adult Drosophila, and we recently reported that a human homolog of the Drosophila Toll protein induces various immune response genes via this pathway. We now demonstrate that signaling by the human Toll receptor employs an adaptor protein, MyD88, and induces activation of NF kappaB via the Pelle-like kinase IRAK and the TRAF6 protein, similar to IL-1R mediated NF-kappaB activation. However, we find that Toll and IL-1R signaling pathways are not identical with respect to AP-1 activation. Finally, our findings implicate MyD88 as a general adaptor/regulator molecule for the Toll/IL-1R family of receptors for innate immunity. PMID- 9734364 TI - A new role for hypoxia in tumor progression: induction of fragile site triggering genomic rearrangements and formation of complex DMs and HSRs. AB - Genome rearrangements including gene amplification are frequent properties of tumor cells, but how they are related to the tumor microenvironment is unknown. Here, we report direct evidence for a causal relationship between hypoxia, induction of fragile sites, and gene amplification. Recently, we showed that breaks at fragile sites initiate intrachromosomal amplification. We demonstrate here that hypoxia is a potent fragile site inducer and that, like fragile sites inducing drugs, it drives fusion of double minutes (DMs) and their targeted reintegration into chromosomal fragile sites, generating homogeneously staining regions (HSRs). This pathway operates efficiently for DMs bearing different sequences, suggesting a model of hypoxia-driven formation of the HSRs containing nonsyntenic sequences frequently observed in solid tumors. PMID- 9734365 TI - High-resolution mapping of crossovers in human sperm defines a minisatellite associated recombination hotspot. AB - Little is known about the fine-scale distribution of meiotic crossovers in human chromosomes. Methods have therefore been developed for detecting and mapping recombination products directly in human sperm DNA. Analysis of crossovers adjacent to the GC-rich minisatellite MS32, which is known to mutate by conversion and crossover within the repeat array, revealed an intense and highly localized recombination hotspot centered upstream of the locus and extending into the beginning of the minisatellite. Allele-specific cosuppression of crossovers and repeat instability suggests that the hotspot is responsible for driving repeat turnover at MS32 and thus that minisatellites might evolve as by-products of localized meiotic recombination in the human genome. PMID- 9734367 TI - PPAR--the good news and the bad. PMID- 9734366 TI - Absence of monocyte chemoattractant protein-1 reduces atherosclerosis in low density lipoprotein receptor-deficient mice. AB - Recruitment of blood monocytes into the arterial subendothelium is one of the earliest steps in atherogenesis. Monocyte chemoattractant protein-1 (MCP-1), a CC chemokine, is one likely signal involved in this process. To test MCP-1's role in atherogenesis, low density lipoprotein (LDL) receptor-deficient mice were made genetically deficient for MCP-1 and fed a high cholesterol diet. Despite having the same amount of total and fractionated serum cholesterol as LDL receptor deficient mice with wild-type MCP-1 alleles, LDL receptor/MCP-1-deficient mice had 83% less lipid deposition throughout their aortas. Consistent with MCP-1 's monocyte chemoattractant properties, compound-deficient mice also had fewer macrophages in their aortic walls. Thus, MCP-1 plays a unique and crucial role in the initiation of atherosclerosis and may provide a new therapeutic target in this disorder. PMID- 9734368 TI - Prospects for live attenuated HIV. PMID- 9734369 TI - Problems with over-the-counter 5-hydroxy-L-tryptophan. PMID- 9734370 TI - Cancer, angiogenesis and fractals. PMID- 9734371 TI - Nattering on about names. PMID- 9734372 TI - Act now on antibiotic resistance. PMID- 9734373 TI - What can South Africa contribute to the AIDS epidemic? PMID- 9734374 TI - UK moves ahead on the xenotransplantation issue. PMID- 9734375 TI - Will Canada establish a national research network? PMID- 9734376 TI - Cardiovascular disease increasing in developing countries. PMID- 9734377 TI - New rating system for UK universities. PMID- 9734378 TI - US healthcare: take a number, please. PMID- 9734379 TI - Japanese officials were aware of HIV in blood products. PMID- 9734380 TI - Brundtland takes charge and restructures the WHO. PMID- 9734381 TI - Who should subsidize NCI clinical trials? PMID- 9734382 TI - US report blasts UNAIDS. PMID- 9734383 TI - Endogenous growth theory for natural scientists. PMID- 9734384 TI - More ado about HIV's origins. PMID- 9734385 TI - Yayoi era mutation disrupts brain and muscle. PMID- 9734386 TI - PPARgamma and colorectal carcinoma: conflicts in a nuclear family. PMID- 9734387 TI - Those that were blind can now see. PMID- 9734388 TI - The art of tolerance. PMID- 9734389 TI - Watching the pot boil. PMID- 9734390 TI - Hematopoietic stem cells: are they CD34-positive or CD34-negative? PMID- 9734391 TI - Alternatives to death: understanding androgen-independent prostate cancer. PMID- 9734392 TI - Killing tumor cells with viruses--a question of specificity. PMID- 9734393 TI - Antigen persistence and time of T-cell tolerization determine the efficacy of tolerization protocols for prevention of skin graft rejection. AB - We studied antigen-specific T-cell tolerization therapy using skin transplantation across a defined minor histocompatibility antigen difference. Specific tolerization protocols using short-lived peptide or long-lived spleen cells presenting the peptide as antigen prevented graft rejection without immunosuppression when started before or as long as 10 days after transplantation. Peptide-induced T-cell tolerance was transient, and antigen presentation by the graft was not sufficient to maintain tolerance. In contrast, transfer of antigen-expressing lymphoid cells induced long-lasting tolerance correlating with donor cell chimerism. These findings show that antigen-specific tolerization can induce graft acceptance even when begun after transplantation and that long-term graft survival depends on persistence of the tolerizing antigen. PMID- 9734394 TI - Defective production of both leukemia inhibitory factor and type 2 T-helper cytokines by decidual T cells in unexplained recurrent abortions. AB - Leukemia inhibitory factor is essential for embryo implantation, and a shift from type 1 T-helper to type 2 T-helper response at the fetal-maternal interface may contribute to successful pregnancy. We show that LIF production is associated with type 2 T-helper cells, is upregulated by IL-4 and progesterone and is downregulated by IL-12, IFN-gamma and IFN-alpha. We also show a decreased production of LIF, IL-4 and IL-10 by decidual T cells of women with unexplained recurrent abortions in comparison with that of women with normal gestation. The defective production of LIF and/or type 2 T-helper cytokines may contribute to the development of unexplained recurrent abortions. PMID- 9734395 TI - Suppression of accelerated diabetic atherosclerosis by the soluble receptor for advanced glycation endproducts. AB - Accelerated atherosclerosis in patients with diabetes is a major cause of their morbidity and mortality, and it is unresponsive to therapy aimed at restoring relative euglycemia. In hyperglycemia, nonenzymatic glycation and oxidation of proteins and lipids results in the accumulation of irreversibly formed advanced glycation endproducts. These advanced glycation endproducts engage their receptor in cells of the blood vessel wall, thereby activating mechanisms linked to the development of vascular lesions. We report here a model of accelerated and advanced atherosclerosis in diabetic mice deficient for apolipoprotein E. Treatment of these mice with the soluble extracellular domain of the receptor for advanced glycation endproducts completely suppressed diabetic atherosclerosis in a glycemia- and lipid-independent manner. These findings indicate interaction between the advanced glycation endproducts and their receptor is involved in the development of accelerated atherosclerosis in diabetes, and identify this receptor as a new therapeutic target in diabetic macrovascular disease. PMID- 9734396 TI - Identification of a new human immunodeficiency virus type 1 distinct from group M and group O. AB - A highly divergent HIV-1 isolate, designated YBF 30, was obtained in 1995 from a 40-year-old Cameroonian woman with AIDS. Depending on the genes studied, phylogenetic analysis showed that YBF30 branched either with SIVcpz-gab or between SIVcpz-gab and HIV-1 group M. The structural genes and tat, vpr, and nef of YBF30 are approximately equidistant from those of HIV-1 group M and SIVcpz gab. In contrast, vif and rev are closer to HIV-1 group M, and vpu is highly divergent. Using a YBF30 V3 loop peptide enzyme immunoassay, we screened 700 HIV 1-positive sera collected in Cameroon; three reacted strongly with the YBF30 peptides and one was confirmed as being related to YBF30 by genetic analysis of a pol fragment. YBF30 is as distinct from SIVcpz-gab as it is from HIV-1 group M and can thus be considered as the prototype strain of a new human immunodeficiency virus group. PMID- 9734397 TI - A newly discovered class of human hematopoietic cells with SCID-repopulating activity. AB - The detection of primitive hematopoietic cells based on repopulation of immune deficient mice is a powerful tool to characterize the human stem-cell compartment. Here, we identify a newly discovered human repopulating cell, distinct from previously identified repopulating cells, that initiates multilineage hematopoiesis in NOD/SCID mice. We call such cells CD34neg-SCID repopulating cells, or CD34neg-SRC. CD34neg-SRC are restricted to a Lin-CD34-CD38 population without detectable surface markers for multiple lineages and CD38 or those previously associated with stem cells (HLA-DR, Thy-1 and CD34). In contrast to CD34+ subfractions, Lin-CD34-CD38- cells have low clonogenicity in short-and long-term in vitro assays. The number of CD34neg-SRC increased in short-term suspension cultures in conditions that did not maintain SRC derived from CD34+ populations, providing independent biological evidence of their distinctiveness. The identification of this newly discovered cell demonstrates complexity of the organization of the human stem-cell compartment and has important implications for clinical applications involving stem-cell transplantation. PMID- 9734398 TI - Differentiation and reversal of malignant changes in colon cancer through PPARgamma. AB - PPARgamma is a nuclear receptor that has a dominant regulatory role in differentiation of cells of the adipose lineage, and has recently been shown to be expressed in the colon. We show here that PPARgamma is expressed at high levels in both well- and poorly-differentiated adenocarcinomas, in normal colonic mucosa and in human colon cancer cell lines. Ligand activation of this receptor in colon cancer cells causes a considerable reduction in linear and clonogenic growth, increased expression of carcinoembryonic antigen and the reversal of many gene expression events specifically associated with colon cancer. Transplantable tumors derived from human colon cancer cells show a significant reduction of growth when mice are treated with troglitazone, a PPARgamma ligand. These results indicate that the growth and differentiation of colon cancer cells can be modulated through PPARgamma. PMID- 9734400 TI - Activators of the nuclear receptor PPARgamma enhance colon polyp formation. AB - A high-fat diet increases the risk of colon, breast and prostate cancer. The molecular mechanism by which dietary lipids promote tumorigenesis is unknown. Their effects may be mediated at least in part by the peroxisome proliferator activated receptors (PPARs). These ligand-activated nuclear receptors modulate gene expression in response to fatty acids, lipid-derived metabolites and antidiabetic drugs. To explore the role of the PPARs in diet-induced carcinogenesis, we treated mice predisposed to intestinal neoplasia with a synthetic PPARgamma ligand. Reflecting the pattern of expression of PPARgamma in the gastrointestinal tract, treated mice developed a considerably greater number of polyps in the colon but not in the small intestine, indicating that PPARgamma activation may provide a molecular link between a high-fat diet and increased risk of colorectal cancer. PMID- 9734399 TI - Activation of the peroxisome proliferator-activated receptor gamma promotes the development of colon tumors in C57BL/6J-APCMin/+ mice. AB - The development of colorectal cancer, one of the most frequent cancers, is influenced by prostaglandins and fatty acids. Decreased prostaglandin production, seen in mice with mutations in the cyclooxygenase 2 gene or in animals and humans treated with cyclooxygenase inhibitors, prevents or attenuates colon cancer development. There is also a strong correlation between the intake of fatty acids from animal origin and colon cancer. Therefore, the peroxisome proliferator activated receptor gamma (PPARgamma), a downstream transcriptional mediator for prostaglandins and fatty acids which is highly expressed in the colon may be involved in this process. Activation of PPARgamma by two different synthetic agonists increased the frequency and size of colon tumors in C57BL/6J-APCMin/+ mice, an animal model susceptible to intestinal neoplasia. Tumor frequency was only increased in the colon, and did not change in the small intestine, coinciding with the colon-restricted expression of PPARgamma. Treatment with PPARgamma agonists increased beta-catenin levels both in the colon of C57BL/61 APCMin/+ mice and in HT-29 colon carcinoma cells. Genetic abnormalities in the Wnt/wingless/APC pathway, which enhance the transcriptional activity of the beta catenin-T-cell factor/lymphoid enhancer factor 1 transcription complex, often underly the development of colon tumors. Our data indicate that PPARgamma activation modifies the development of colon tumors in C57BL/61-APCMin/+ mice. PMID- 9734401 TI - Suppression of caveolin expression induces androgen sensitivity in metastatic androgen-insensitive mouse prostate cancer cells. AB - Although prostate cancer cells are often initially sensitive to androgen ablation, they eventually lose this response and continue to survive, grow and spread in the absence of androgenic steroids. The mechanism(s) that underlie resistance to androgen ablation therapy remain mostly unknown. We have demonstrated that elevated caveolin protein levels are associated with human prostate cancer progression in pathological specimens. Here we show that suppression of caveolin expression by a stably transfected antisense caveolin-1 cDNA vector converted androgen-insensitive metastatic mouse prostate cancer cells to an androgen-sensitive phenotype. Orthotopically grown tumors and low-density cell cultures derived from antisense caveolin clones had increased apoptosis in the absence of androgenic steroids, whereas similarly grown tumors and cells from vector (control) clones and parental cells were not sensitive to androgens. Studies using a representative antisense caveolin clone showed that selection for androgen resistance in vivo correlated with increased caveolin levels, and that adenovirus-mediated caveolin expression blocked androgen sensitivity. Our results identify a new candidate gene for hormone-resistant prostate cancer in man and indicate that androgen insensitivity can be an inherent property of metastatic prostate cancer. PMID- 9734402 TI - The core protein of hepatitis C virus induces hepatocellular carcinoma in transgenic mice. AB - Hepatitis C virus (HCV) is the main cause of chronic hepatitis worldwide. Chronic hepatitis ultimately results in the development of hepatocellular carcinoma (HCC). However, the mechanism of hepatocarcinogenesis in chronic HCV infection is still unclear. The ability of the core protein of HCV to modulate gene transcription, cell proliferation and cell death may be involved in the pathogenesis of HCC. Here, we report the development of HCC in two independent lines of mice transgenic for the HCV core gene, which develop hepatic steatosis early in life as a histological feature characteristic of chronic hepatitis C. After the age of 16 months, mice of both lines developed hepatic tumors that first appeared as adenomas containing fat droplets in the cytoplasm. Then HCC, a more poorly-differentiated neoplasia, developed from within the adenomas, presenting in a 'nodule-in-nodule' manner without cytoplasmic fat droplets; this closely resembled the histopathological characteristics of the early stage of HCC in patients with chronic hepatitis C. These results indicate that the HCV core protein has a chief role in the development of HCC, and that these transgenic mice provide good animal models for determining the molecular events in hepatocarcinogenesis with HCV infection. PMID- 9734403 TI - p53-dependent cell death/apoptosis is required for a productive adenovirus infection. AB - The p53 tumor suppressor protein binds to both cellular and viral proteins, which influence its biological activity. One such protein is the large E1b tumor antigen (E1b58kDa) from adenoviruses (Ads), which abrogates the ability of p53 to transactivate various promoters. This inactivation of p53 function is believed to be the mechanism by which E1b58kDa contributes to the cell transformation process. Although the p53-E1b58kDa complex occurs during infection and is conserved among different serotypes, there are limited data demonstrating that it has a role in virus replication. However, loss of p53 expression occurs after adenovirus infection of human cells and an E1b58kDa deletion mutant (Onyx-015, also called dl 1520) selectively replicates in p53-defective cells. These (and other) data indicate a plausible hypothesis is that loss of p53 function may be conducive to efficient adenovirus replication. However, wild-type (wt) Ad5 grows more efficiently in cells expressing a wt p53 protein. These studies indicate that the hypothesis may be an oversimplification. Here, we show that cells expressing wt p53, as well as p53-defective cells, allow adenovirus replication, but only cells expressing wt p53 show evidence of virus-induced cytopathic effect. This correlates with the ability of adenovirus to induce cell death. Our data indicate that p53 plays a necessary part in mediating cellular destruction to allow a productive adenovirus infection. In contrast, p53-deficient cells are less sensitive to the cytolytic effects of adenovirus and as such raise questions about the use of E1b58kDa-deficient adenoviruses in tumor therapy. PMID- 9734404 TI - Expression of heme oxygenase-1 can determine cardiac xenograft survival. AB - The rejection of concordant xenografts, such as mouse-to-rat cardiac xenografts, is very similar to the delayed rejection of porcine-to-primate discordant xenografts. In concordant models, this type of rejection is prevented by brief complement inhibition by cobra venom factor (CVF) and sustained T-cell immunosuppression by cyclosporin A (CyA). Mouse hearts that survive indefinitely in rats treated with CVF plus CyA express the anti-inflammatory gene heme oxygenase-1 (HO-1) in their endothelial cells and smooth muscle cells. The anti inflammatory properties of HO-1 are thought to rely on the ability of this enzyme to degrade heme and generate bilirubin, free iron and carbon monoxide. Bilirubin is a potent anti-oxidant, free iron upregulates the transcription of the cytoprotective gene, ferritin, and carbon monoxide is thought to be essential in regulating vascular relaxation in a manner similar to nitric oxide. We show here that the expression of the HO-1 gene is functionally associated with xenograft survival, and that rapid expression of HO-1 in cardiac xenografts can be essential to ensure long-term xenograft survival. PMID- 9734405 TI - Impaired blood-brain barrier function in angiotensinogen-deficient mice. AB - Astrocytes in the central nervous system have physiologically important roles in the response to brain injury. Brain damage results in disruption of the blood brain barrier (BBB), producing detachment of astrocyte endfeet from endothelial cells. The resultant leakage of serum proteins from loosened tight junctions between endothelial cells produces brain edema. At the same time, reactive astrocytes migrate to the injured area, where they proliferate and produce extracellular matrix, thereby reconstituting the BBB. As astrocytes are known to express angiotensinogen, which is the precursor of angiotensins (AI to AIV), we have investigated a possible functional contribution of angiotensinogen or one of its metabolites to BBB reconstitution. The astrocytes of angiotensinogen knockout mice had very attenuated expression of glial fibrially acidic protein and decreased laminin production in response to cold injury, and ultimately incomplete reconstitution of impaired BBB function. Although these abnormalities were rescued by administration of AII or AIV, the restoration of BBB function was not inhibited by AII type 1 and 2 receptor antagonists. These findings provide evidence that astrocytes with angiotensins are required for functional maintenance of the BBB. PMID- 9734406 TI - Computer-based training for the treatment of partial blindness. AB - Partial blindness after brain injury has been considered non-treatable. To evaluate whether patients with visual-field defects can profit from computer based visual restitution training (VRT), two independent clinical trials were conducted using patients with optic nerve (n = 19) or post-chiasmatic brain injury (n = 19). In post-chiasma patients, VRT led to a significant improvement (29.4%) over baseline in the ability to detect visual stimuli; in optic nerve patients, the effects were even more pronounced (73.6% improvement). Visual-field enlargements were confirmed by the observation of a visual-field expansion of 4.9 degrees-5.8 degrees of visual angle and improved acuity in optic nerve patients. Ninety five percent of the VRT-treated patients showed improvements, 72.2% confirmed visual improvements subjectively. Patients receiving a placebo training did not show comparable improvements. In conclusion, VRT with a computer program improves vision in patients with visual-field defects and offers a new, cost effective therapy for partial blindness. PMID- 9734407 TI - One-stage transfer of the latissimus dorsi muscle for reanimation of a paralyzed face: a new alternative. AB - The two-stage method combining neurovascular free-muscle transfer with cross-face nerve grafting is now a widely accepted procedure for dynamic smile reconstruction in cases with long established unilateral facial paralysis. Although the results are promising, the two operations, about 1 year apart, exert an economic burden on the patients and require a lengthy period before obtaining results. Sequelae such as hypoesthesia, paresthesia, and conspicuous scar on the donor leg for harvesting a sural nerve graft also cannot be disregarded. To overcome such drawbacks of the two-stage method, we report a refined technique utilizing one-stage microvascular free transfer of the latissimus dorsi muscle. Its thoracodorsal nerve is crossed through the upper lip and sutured to the contralateral intact facial nerve branches. Reinnervation of the transferred muscle is established at a mean of 7 months postoperatively, which is faster than that of the two-stage method. In our present series with 24 patients, 21 patients (more than 87 percent) believed that their results were excellent or satisfactory, which also compares well with the results of the two-stage method combining free-muscle transfer with cross-face nerve graft. PMID- 9734408 TI - Functional outcome after surgery for trigonocephaly. AB - The long-term mental outcome of 76 children operated on for trigonocephaly was assessed, and the factors influencing the prognosis were studied. Final assessment of mental development was made on children who were more than 3 years old and was based on the occurrence of behavioral disturbances, learning disability, and school difficulties, and on intellectual efficiency. Children were graded into three groups: no abnormality, mild abnormalities but with normal social function, and grossly abnormal. Preoperative computed tomography scans were used to measure the severity of the frontal stenosis and to identify associated intracranial abnormalities, such as agenesis of the corpus callosum, dilatation of the subdural spaces, or hydrocephalus. Associated extracranial malformations and associated family cases were also noted. Lastly, the family setting was studied. Overall, 31.6 percent of patients had evidence of some degree of trouble. Several correlations were identified: mental development was worse when the frontal stenosis was severe, when cranial reconstruction was performed after 1 year of age, and when there were associated extracranial malformations. In addition, the family environment was found to have a major influence, but the presence of intracranial abnormalities did not correlate with mental development. PMID- 9734409 TI - Mobius syndrome: classification and grading system. AB - Mobius syndrome is characterized by facial abnormalities, but the limbs, chest wall, spine, and soft tissues also can be involved. There is no system for categorizing the various anomalies, grading phenotypic severity, designing treatment protocols, or assessing therapeutic results. This is a retrospective analysis of 27 patients with Mobius syndrome seen in our craniofacial unit from 1980 to 1994. We categorized and graded the cranial nerve deficits and diverse musculoskeletal abnormalities of the face, upper and lower limbs, and trunk. The first letter for each of five potentially involved structures, i.e., cranial nerve, lower limb, upper limb, face, and thorax, formed the acronym CLUFT. The structural and/or functional deficits for each component were graded on a scale of 0 to 3. Complete facial nerve paralysis was documented in 11 patients and paresis in 16. Facial nerve paralysis was bilaterally symmetric in 17 of 26 patients. Sixth nerve paralysis was present in 23 of 27 patients; other cranial nerves were affected in 8 of 27 patients. Lower limbs were involved in 10 of 27 patients and upper limbs in 7 of 27 patients. Facial structures were affected in 17 of 27 patients (e.g., microtia, micrognathia, and microphthalmia), and chest wall deformities were found in 8 of 27 patients (e.g., scoliosis, hypoplasia of the breast, pectoral muscles, and scapula). We noted that microtia primarily involved second pharyngeal arch-derived structures. The CLUFT system permits categorization and comparison of Mobius patients for phenotypic and management outcome studies. Documentation of the widespread structural anomalies suggests that pathogenesis involves vascular disruption; a detailed prenatal history is indicated. PMID- 9734410 TI - Mitochondrial activity of orbicularis oris muscle in unilateral cleft lip patients. AB - To better evaluate the role of a possible mitochondrial alteration in the pathogenesis of cleft lip, we obtained and examined 38 orbicularis oris muscle specimens taken from the cleft margin of both cleft and noncleft sides of 10 unilateral cleft lip infants at the time of primary closure. Part of each sample was frozen in liquid nitrogen/cooled isopentane, while the remainder was fixed in 2.5% glutaraldehyde, postfixed in osmium tetroxide, and embedded in Araldyte resin. Ten-micrometer-thick sections were obtained from the frozen samples and stained for histologic (Gomori trichrome) and histochemical (adenosine triphosphatase, nicotinamide adenine dinucleotide-tetrazolium reductase, cytochrome c-oxidase, succinate dehydrogenase) techniques. Ultra-thin sections (70 to 100 nm) of the resin-embedded specimens were stained with uranyl acetate and lead cytrate and were examined with a Zeiss 109 transmission electron microscope operating at 80 kV. Muscular fiber-type ratio was found to be 19.2 percent type 1 and 80.8 percent type 2 fibers on the cleft side and 26.3 percent type 1 and 73.7 percent type 2 fibers on the noncleft side. We detected aspecific structural alterations, such as variations in the fiber size without fiber group atrophy or fiber-type grouping with the ATPase reaction, in all biopsies. Although Gomori trichrome revealed a dark staining and red granularity of the fibers, suggesting an increase in mitochondria activity, no ragged-red fibers or cytochrome c-oxidase-negative/succinate dehydrogenase-positive fibers were found. At the ultrastructural level, the mitochondrial morphology was always preserved, without inclusions or variations in size and/or shape. On the other hand, we invariably noticed an increase of the number of mitochondria, associated with abnormal glycogen deposits, in some areas of every specimen. Both of these two latter findings were regularly localized at the periphery of the sarcolemma, resembling the so-called lobulated fibers, an aspecific sign of muscular flogosis. Our findings, although excluding an inherent metabolic myopathy of orbicularis oris muscle in unilateral cleft lip patients, evinced both an increased oxidative metabolism and a generic inflammatory condition of that muscle, the nature of which must still be defined. PMID- 9734411 TI - Management of displaced lateral orbital wall fractures associated with visual and ocular motility disturbances. AB - Impacted fractures of the lateral orbital wall are a type of orbital blow-in fracture that may be accompanied by decreased visual acuity and ocular motility limitations. Eleven patients who suffered this injury triad were retrospectively reviewed to determine the nature of the ophthalmologic injuries and the effect of fracture reduction on recovery of ophthalmologic functions. Two patients with decreased visual acuity owing to trauma to the globe recovered to subjective pretrauma levels following surgery. Nine patients were thought to have a traumatic optic neuropathy with varying degrees of visual loss. Patients with an injury to the intraorbital portion of the optic nerve and a presurgical visual acuity of 20/400 or better recovered to subjective pretrauma levels. Those with visual acuity of less than 20/400 or an injury to the intracanalicular portion of the nerve had responses ranging from no improvement to objective improvement with large field defects. Ocular motility improved in all patients, many in the immediate postsurgical period consistent with removal of a mechanical restriction. No patients had worsening of ophthalmologic deficits as a result of manipulation of fracture fragments. Our experience suggests that early surgical intervention facilitates recovery of vision and eye movement. The traumatic optic neuropathy that accompanies this fracture is distinct from the indirect type of optic nerve injury that may respond to steroids, and the ophthalmoplegia is distinct from the usual traumatic superior orbital fissure syndrome that resolves spontaneously. An understanding of the impacted lateral orbital wall fracture and its ophthalmologic implications is essential for any surgeon who desires to manage craniomaxillofacial injuries. PMID- 9734412 TI - Eye socket reconstruction with the prefabricated temporal island flap. AB - Insufficiency of tissues and progressive contraction usually restrict the application of prosthetic devices in anophthalmic eye sockets. To achieve a successful reconstruction, the plastic surgeon has to form a socket that has proper dimensions and is completely covered by a well vascularized epithelial surface. Eye socket reconstruction with free skin, mucous membrane, or dermis-fat grafts usually remains unsatisfactory in severe cases. We have used a prefabricated temporal island flap to solve this difficult problem since 1983. In this method, a full-thickness skin graft is applied over the temporal fascia to create a prefabricated island flap based on the superficial temporal vessels. This flap is transposed into the eye socket 3 weeks later. Some modifications in flap design have been done to get better fitting of the prosthesis since that time. Thirty-three patients with constricted eye sockets that could not use prosthetic devices were treated with prefabricated temporal island flaps since 1983. The follow-up period was between 1 and 13 years. Eye sockets with adequate size and volume were created in all patients, and the results were successful. This method prevented secondary graft shrinkage, and the prefabricated island flaps preserved their dome shape during the follow-up period. We believe this method is a useful one in the treatment of the contracted socket. PMID- 9734413 TI - Reconstruction of defects involving the upper one-third of the auricle. AB - We describe here a reconstructive technique for the repair of defects involving the upper one-third of the auricle using a combination of several flaps. For a defect of the preauricular surface only, a chondrocutaneous flap from the concha and a postauricular subcutaneous pedicle flap were used. For a full-thickness defect, the reconstruction was made by using the chondrocutaneous flap, the postauricular subcutaneous pedicle flap, and a postauricular skin flap. The final skin defect was collected in the postauricular region and was directly sutured or covered by skin grafting. Seven patients were treated by this procedure. Although the tip of the postauricular skin flap fell into partial necrosis in one patient, all of the other flaps were well healed without vascular stasis. All of the reconstructed auricles had good three-dimensional forms with satisfactory color and texture to match. There was neither constriction of the ear nor deformation owing to scar contracture. PMID- 9734414 TI - Functional reconstruction of the tongue and deglutition muscles following extensive resection of tongue cancer. AB - The authors describe their experience with functional restoration of tongue and deglutition muscles at the floor of the mouth after an extensive resection of tongue cancer. Five patients underwent immediate tongue reconstruction using a reinnervated rectus abdominis myocutaneous free flap in which the included tenth intercostal nerve was coapted to the remaining hypoglossal nerve. The rectus sheath strips attached on both cut ends of the muscle were used to create the firm tendinous insertions between the mandible and hyoid bone based on the anatomic findings of the extrinsic tongue and suprahyoid muscles. The postoperative course was uneventful in all patients. All patients presented with good tongue bulk without obvious atrophy. Three patients with subtotal glossectomy demonstrated good cooperative mobility of the reconstructed and remaining tongue and had solid or semisolid/soft diet. However, two patients with total glossectomy did not show satisfactory rehabilitation of the reconstructed tongue. Postoperative electromyographic assessment in two patients showed good functional recovery of the grafted muscle. The cine-magnetic resonance imaging deglutition study in one patient with 80-percent tongue resection demonstrated sufficient elevation of the dorsal base of the reconstructed tongue, contraction of the reconstructed deglutition muscles, complete glossopalatal closure, and elevation of the hyoid bone and larynx during the deglutition. This reconstructive technique is strongly recommended for the patients who have undergone subtotal glossectomy to provide physiological functional recovery of the reconstructed tongue synchronizing with the remaining tongue. PMID- 9734415 TI - Double opposing semicircular flap: a modification of opposing Z-plasty for closing circular defects. AB - A new method is presented for the closure of circular skin defects without altering their original shapes and excision of additional healthy skin. The authors hypothesized that two semicircular flaps prepared from opposite sides of a circular defect could close the defect without additional healthy tissue excision from the defect or the raised flaps; this technique was called double opposing semicircular flaps. It was first studied on an animal model followed by application on our patients with satisfactory results. This procedure has been performed with success on various body surfaces for defects of a size range of 2 to 5 cm in diameter. In this article, technical details and four clinical cases are presented. PMID- 9734416 TI - Tissue expander complications in the pediatric burn patient. AB - Tissue expanders have become a useful adjuvant in pediatric burn reconstruction. We reviewed our experience with tissue expanders from June of 1984 to July of 1995. There were 403 expanders used in 301 patients. Complications relative to specific anatomic areas from July of 1987 to July of 1995 were compared with previously published data in the journal from June of 1984 to June of 1987. Complications were defined as absolute if they resulted in the loss of expanders or in additional surgery, or none of the preoperative plan was satisfied. The relative complications were defined as spotty alopecia, alopecia greater than 50 percent, or the operative plan was only partially satisfied, sometimes implying poor surgical judgment. The overall complication rate for the period June of 1984 to June of 1987 was 30 percent (37 complications in 122 expanders). In the July of 1987 to July of 1995 study, the complication rate was only 18 percent (51 complications in 281 expanders). This was a statistically significant decrease between the periods (p = 0.010). In the recent 8-year period, there was a decrease compared with the previous study in both the absolute and relative complications. The most common absolute complication in this period was infection (15 of 31, 48 percent) with 12 (39 percent) being early infection. With regard to the nine complications in the neck, face, ear, and supraclavicular area, two thirds were related to leakage or exposure of the expanders, resulting from the tight anatomic area causing mechanical damage of the expanders as well as ischemia to the overlying skin. Early in the study, the lower extremities proved to involve difficult or unsatisfactory areas to expand, and lower extremity expansion was abandoned throughout the remainder of the study period. The overall decrease in absolute and relative complications is likely the result of increased operative experience as well as a developed protocol for the prevention of perioperative complications relating to infection and expansion in high-risk anatomic sites. PMID- 9734417 TI - Early excision and grafting versus conservative management of burns in the elderly. AB - Elderly burn patients have significantly higher mortality rates than younger patients with similar burns over the total body surface area. Two theories exist regarding treatment of burns in the elderly: a traditional approach to limit physiologic stress by avoidance of operative intervention in the early post-burn stage and eschar excision and wound closure within the first week of hospitalization. We examined retrospectively the outcome in patients 70 years or older, hospitalized in the University of Kentucky Burn Unit between 1975 and 1995. In the first decade (1975 to 1983), patients were managed conservatively, namely, with spontaneous eschar separation and late skin grafting. In the second half of the study period (1984 to 1994), elderly patients were managed by early operative excision (<7 days) and grafting. A total of 73 elderly patients were admitted to the unit, 6 of whom were not resuscitated and died shortly (<96 hours) after admission. Twenty-eight patients had early excision and grafting (average age 78.1 years, total body surface area 23.6 percent), and 39 were managed conservatively (average age 79.3 years, total body surface area 20.9 percent). The mortality rate was 57 percent in the first group and 41 percent in the second group (p = 0.22). In an effort to further define the two groups, the other patient variable that contributes to burn mortality besides age and total body surface area, inhalation injury, was subtracted and the mortality rates were recalculated. Excluding patients with inhalation injury, the mortality rate was 48 percent in the first group and 27 percent in the second group (p = 0.15). We conclude that, in our unit, the management of elderly patients by early excision and grafting was of no benefit and may have resulted in a higher mortality rate. PMID- 9734418 TI - Toxic epidermal necrolysis syndrome: mortality rate reduced with early referral to regional burn center. AB - Toxic epidermal necrolysis syndrome is an uncommon, acute, life-threatening disorder that involves sloughing of skin at the dermal-epidermal junction with associated mucositis. Between 1985 and 1995, 36 patients were treated for toxic epidermal necrolysis syndrome, at the Baltimore Regional Burn Center. A retrospective chart analysis was performed to discover significant determinants of mortality. Ninety-seven percent of the patients (35 of 36) were referred from outside institutions after an average of 6.3 +/- 0.8 days. Analysis of the data shows that patients who survived had been referred 7.5 days earlier than nonsurvivors (4.0 +/- 0.5 days versus 11.5 +/- 1.4 days, p < 0.001). When the patients were separated into two groups on the basis of time of referral, those referred "early" (< or = 7 days) had a mortality rate of 4 percent (1 of 24) versus 83 percent (10 of 12) for those referred "late" (> 7 days) (p < 0.001). Data were available from transferring institutions for 21 of the 36 patients. Analysis of the microbiologic data from these 21 patients revealed bacteremia, and subsequent death occurred in 100 percent (6 of 6) of the patients referred with positive cultures, whereas bacteremia developed in only 33 percent (5 of 15) of the patients referred with negative cultures, for a mortality rate of 7 percent (1 of 15). In addition, 86 percent (6 of 7) of the patients who were referred late (> 7 days) had positive cultures on referral. The current trend toward prolonged treatment in outside facilities before referral to a burn center is detrimental to the care of patients with toxic epidermal necrolysis syndrome. The overall rate of bacteremia, septicemia, and mortality is significantly reduced with early (< or = 7 days) referral to a regional burn center. PMID- 9734419 TI - Interleukin-1alpha and collagenase activity are elevated in chronic wounds. AB - Interleukin-1-alpha (IL-1alpha) is a member of a family of proinflammatory polypeptide mediators that has been shown in vitro to stimulate collagenase production. Collagenase is a proteolytic enzyme classified as one of the matrix metalloproteinases (MMP-1) that specifically recognizes and cleaves collagen. Therefore, the objective of this study was to compare the levels of these two proteins in chronic wounds as possible factors in the pathogenesis of chronic wounds. Fluids from 10 chronic wounds were collected before and after a 1-week treatment with a hydroactive dressing (Cutinova cavity). In addition, fluids were collected from 20 acute wounds for comparison. IL-1alpha and MMP-1 levels were quantified using sandwich ELISA. Collagenase activity was measured using a radiolabeled collagen as substrate. Clinically, the chronic wounds showed decreased area (-21.0 cm2) and reduced volume (-134.5 cm3) by 4 weeks after treatment with the hydroactive dressing. There were no significant differences in the protein concentrations between acute wound fluids (21.0 +/- 3.0 mg/ml) and chronic wound fluids before and after treatment with the hydroactive dressing (18.3 +/- 5.5 and 25.2 +/- 7.6 mg/ml, respectively). Levels of IL-1alpha in the acute wound fluids were low (0.019 pg/mg), whereas in the chronic wound fluid before treatment they had been significantly elevated (44.9 + 21.8 pg/mg). Following treatment with the hydroactive dressing, the IL-1alpha levels dropped to 10.3 + 3.3 pg/mg (p < 0.05). Collagenase activity was not detectable in acute wound fluid, elevated in pretreatment chronic wounds (12.9 + 3.4 units), and decreased in chronic wounds after treatment (11.4 + 3.3 units). This study correlated clinical healing of chronic wounds with biochemical changes in the ulcer microenvironment. As the chronic wounds began to heal, there was a significant decrease in the IL-1alpha levels and collagenase activity, thus suggesting that these two proteins may contribute to the lack of healing characteristic of chronic wounds. PMID- 9734420 TI - The intradermal anatomy of the inframammary fold. AB - The anatomy of the inframammary fold has been a subject of controversy. This report describes the anatomic location and the histologic structure of the inframammary fold on the basis of caderveric dissections and microscopic examination. Ten breast cadaver dissections were performed on female cadavers (ages 35 to 72). Twenty specimens after en bloc resections of the inframammary fold and subcutaneous tissue, including the pectoralis muscle, were harvested. Specimens were examined for gross collagen stricture by using India ink to highlight the collagenous aspects of the subcutaneous soft-tissue networks. The inframammary fold skin and dermis from the contralateral breast and control samples of skin and dermis from the upper chest and the abdomen were collected for microscopic studies. These samples were stained with Sirius red and examined microscopically by polarized light. On histologic examination, regular arrays of collagen were found running parallel with the inframammary fold, and the control sections showed random patterns of collagen deposition. On gross examination, a condensation of the superficial fascial system was observed. This formed a zone of adherence between the skin and the underlying pectoralis fascia. The conclusion of this study is that the inframammary fold is an intrinsic dermal structure consisting of regular arrays of collagen held in place by a zone of adherence that is a specialized area of the superficial fascial system. The clinical significance of this study is that the intradermal structure of the inframammary fold should be preserved in any breast procedure for natural aesthetic results. PMID- 9734421 TI - Focus on the breast fascial system: a new approach for inframammary fold reconstruction. AB - Anatomists and surgeons have underestimated the importance of understanding the anatomic connective frame of the inframammary region. The submammary fold does not originate as a self-governing unit but depends on breast mould and on a fine superficial fascial system suspension. The authors investigated the inframammary fold anatomy and subcutaneous breast territory in cadaver and live dissection with histologic analyses, without sharing the theories about superficial fascia splitting and inframammary ligament existence. The authors have understood that a reliable and fine correction of inframammary fold contour in breast reconstruction may only be achieved by an empirical surgical procedure that exclusively concerns the restoration of the superficial fascial system. The literature on this subject is reviewed. Fascial anchoring surgery, after capsulotomy and superficial fasciotomy, without lower thoracic advancement flap or deep subcutaneous undermining, was performed for 100 breast reconstructions after biodimensional device programming. Technique and results are also discussed. PMID- 9734422 TI - Light and electron microscopic evaluation of the pectoralis major muscle following tissue expansion for breast reconstruction. AB - The placement of tissue expanders under the pectoralis major muscle has become a common procedure in breast reconstruction after mastectomy. Little information is available regarding the changes caused by tissue expansion on human skeletal muscle. In this study, we report the light and electron microscopic changes observed in 20 expanded pectoralis major muscles from 12 patients undergoing two stage breast reconstruction procedures. Standard 400-cc round Radovan tissue expanders were placed, eight bilaterally and four unilaterally. Three biopsies were taken from each muscle at different locations during both the first stage (pre-expansion) and 23 weeks later at the second stage (postexpansion). The operative procedures and expansion protocol were the same in all reconstructions. No postoperative radiation therapy was added. The histologic changes were reported in a blind fashion by one pathologist. Light microscopy did not show significant pathologic changes. All but one of the pre-expansion specimens examined by electron microscope were reported as normal, whereas all of the postexpansion biopsies were grossly altered. Focal muscle fiber degeneration with glycogen deposits and mild interstitial fibrosis was noted. In addition, some fibers showed disorganization of the myofilaments in the sarcomeres. The above ultrastructural changes are significant morphologic alterations, which may be the result of muscle hypoxia. Whether these changes indicate permanent or transient transformation is yet unclear. Patient follow-up did not reveal any functional muscular deficit. We conclude that there is definite evidence to suggest significant muscular structural damage after routine subpectoral expansion for breast reconstruction. PMID- 9734423 TI - Paraumbilical perforator flap without deep inferior epigastric vessels. AB - With the introduction of supramicrosurgery, a new paraumbilical perforator flap without a deep inferior epigastric vessel and with very small perforator anastomoses was used for nine patients. The abdominal defects of two patients, the lower leg or foot defects of five patients, and the scalp defects of two patients were repaired with an island perforator flap. The advantages of the paraumbilical perforator flap are as follows: (1) there is a very short operating time for flap elevation; (2) there is no invasion or sacrifice of any rectus abdominis muscle; (3) for middle-aged, obese patients, the donor site may be the best from the cosmetic point of view; (4) many small recipient vessels to anastomose the perforator exist throughout the body; (5) a thin skin flap with adequate thickness can be created easily with simultaneous removal of fatty tissue; (6) secondary defatting around the perforator can be done by minor surgery under local anesthesia; and (7) a vascularized adiposal flap with adequate thickness can be created easily. This flap seems to be indicated for female patients with defects in the abdominal wall and the lower leg. The island flap can easily resurface abdominal skin defects, such as intestinal fistula or radiation ulcers. The free flap is suitable for covering defects in the lower leg, foot, and scalp temporarily before administration of a tissue expander. PMID- 9734424 TI - Liposuction combined with controlled compression therapy reduces arm lymphedema more effectively than controlled compression therapy alone. AB - Arm lymphedema after breast cancer therapy has been treated with various forms of conservative and surgical treatment during recent years. The clinical results usually have been modest or, in some instances, even disappointing. In a previous series of patients treated with the new liposuction technique combined with controlled compression therapy, we found, however, an overall edema reduction of 106 percent after 1 year. The purpose of this study was both to investigate how much the surgical procedure contributes to the outcome and to clarify the importance of controlled compression therapy. Twenty-eight patients were, therefore, prospectively matched into two groups. One group received liposuction combined with controlled compression therapy, and one group received the therapy alone. Additionally, the therapy group was compared with our complete group of patients treated thus far with liposuction combined with therapy (n = 30). The prospective study using matched pairs (n = 14) showed that liposuction combined with controlled compression therapy is significantly more effective than the therapy alone (p < 0.0001), with a mean difference of about 1000 ml during the entire 1-year observation period. The beneficial effect of liposuction was confirmed by the comparison between the controlled compression therapy group and our complete group of patients treated with liposuction combined with the therapy, as the edema reduction figures after 1 year were 47 percent and 104 percent, respectively (p < 0.0001). In six patients who had surgery and a complete reduction of the edema, the compression garments were removed for 1 week, 1 year postoperatively. A marked increase in the arm volume was observed, which was immediately remedied by reapplying the garments. We conclude that liposuction combined with controlled compression therapy reduces arm lymphedema more efficiently than the therapy alone. Continued use of compression garments is, however, important to maintain the primary surgical outcome. PMID- 9734425 TI - An algorithm for early aggressive treatment of frostbite with limb salvage directed by triple-phase scanning. AB - Frostbite injuries have traditionally been treated with expectant observation. With the exception of early blister aspiration tissues are allowed to demarcate before definitive debridement is accomplished. Triple-phase bone scanning has been used to define the extent of fatally damaged tissues in an attempt to allow for early debridement and wound closure. We suggest extending this technology to assess injury and direct debridement in patients for whom early aggressive salvage attempts are indicated. We present two cases in which triple-phase scanning was used to direct early debridement for aggressive limb salvage with flap reconstruction. Bone, ligament, tendon, and nerve were preserved and covered with vascularized tissue before the onset of frank necrosis. Postoperative scans reveal revascularization of these tissues. An algorithm incorporating triple phase scanning for the evaluation and treatment of frostbite is presented. PMID- 9734426 TI - The prepuce free flap: dissection feasibility study and clinical application of a super-thin new flap. AB - The aim of this study was to develop a free flap of the male prepuce. First, a dissection feasibility study was performed in eight male cadavers. Dissection proved feasible, and the mean surface area of the prepuce, when folded out, was 46.7 cm2. The mean pedicle length was 15 cm, with a mean diameter of the inferior external pudendal artery at its origin of 1.2 mm and a mean diameter of the inferior external pudendal vein at its origin of 1.9 mm. Next, a viability study was performed by isolating a prepuce free flap during male-to-female sex reassignment procedures. The flap as well as the residual skin of the penis shaft remained well perfused. Thereafter, the new prepuce free flap was successfully applied for the reconstruction of defects of the floor of the mouth, tongue, and oropharynx in two patients. The thinness and pliability of the flap, its large surface area, the long vascular pedicle, and the limited donor-site morbidity are the major advantages of this new flap. The possibility to raise the flap simultaneously with the tumor resection is an additional advantage. The small caliber of the flap artery seems to be the only drawback. PMID- 9734427 TI - The current role of preoperative arteriography in free fibula flaps. AB - The free fibula flap has become a "workhorse" flap for reconstructive surgeons, yet the indications for preoperative arteriography for the donor extremity remain unclear. Therefore, a retrospective review of all free fibula candidates over a 4 year period was conducted to clarify the need for preoperative arteriography. One hundred consecutive patients were evaluated as potential candidates for free fibula reconstruction. Twenty-one patients were deemed unsuitable because of associated comorbid conditions (15) or unusable limbs (6). The remaining 79 patients were candidates for fibula free flap reconstruction. Eight patients (10 percent) who had an abnormal lower extremity vascular physical examination (diminished or absent pedal pulses) underwent arteriography to evaluate the fibula donor site. Free fibula transfer was performed in 77 patients (mean age, 41; range, 3 to 80 years) to the following sites: mandible (65), upper limb (4), lower limb (6), and trunk (2). The overall free flap success rate was 99 percent. Results of arteriography included: normal three-vessel runoff (6), bilateral peroneal arteria magna (1), and bilateral posterior tibial artery occlusion with reconstitution via the peroneal artery (1). Two patients with unusable fibula donor sites (determination based on arteriographic findings) were reconstructed with ilium and radius. All others underwent uncomplicated free fibula transfer. Ischemic complications at the fibula donor site did not occur in any patient. This study supports the use of lower extremity vascular physical examination as the primary means of evaluating the fibula donor site. Routine preoperative arteriography is unnecessary and should be reserved for those patients with abnormal vascular examinations. PMID- 9734428 TI - Endoscopic versus open carpal tunnel release: a cost-effectiveness analysis. AB - Endoscopic carpal tunnel release is a controversial procedure used in the treatment of carpal tunnel syndrome. Although endoscopic carpal tunnel release is associated with less incisional pain and faster recovery time than the open carpal tunnel release, opponents of endoscopic carpal tunnel release suggest that its benefits are outweighed by its higher complication rates from median nerve transection and transient numbness of the fingers. Because of the huge economic and social impact of carpal tunnel syndrome in this country, we performed a cost effectiveness analysis comparing endoscopic carpal tunnel release and open carpal tunnel release using guidelines established by the Panel on Cost-Effectiveness in Health and Medicine of the U.S. Public Health Service. A decision analytic model was used to measure differences in cost and effectiveness--expressed as quality adjusted life-years (QALYs)--between endoscopic carpal tunnel release and open carpal tunnel release. The societal perspective was chosen, and probabilities for various outcomes for the two procedures were obtained from published randomized controlled trials. Cost data were derived from the Medicare Resource-Based Relative Value Units published in the Federal Register. QALYs were obtained from two groups of health care providers using a utility-assessment questionnaire. Using probabilities for various outcomes from the two published randomized controlled trials comparing endoscopic carpal tunnel release and open carpal tunnel release, we constructed a decision tree to derive both the cost and the QALYs for the two procedures. The incremental cost difference between endoscopic carpal tunnel release and open carpal tunnel release was $46, using Medicare cost and probabilities of various outcomes derived from a study by Brown et al. in 1993. We calculated QALYs for five age groups--25, 35, 45, 55, 65--assuming a life expectancy of 75 years. The marginal effectiveness (QALY of endoscopic carpal tunnel release minus QALY of open carpal tunnel release) ranged from 0.235 QALY for the 25-year-old age group to 0.066 QALY for the 65-year-old age group, giving a cost-effectiveness ratio of $195/QALY and $693/QALY, respectively. When compared with other accepted medical interventions such as breast cancer screening ($4836/QALY) and exercise to prevent coronary heart disease ($13,508/QALY), endoscopic carpal tunnel release seems to be cost-effective. However, our sensitivity analysis indicated that the cost-effectiveness ratio was very sensitive to a major complication such as median nerve injury. For endoscopic carpal tunnel release to be a cost-effective procedure, the incidence of median nerve injury must be one percentage point less for endoscopic carpal tunnel release than for open carpal tunnel release. Based on the data from the randomized-controlled trials, endoscopic carpal tunnel release seems to be a cost effective procedure; however, before it can be recommended, greater emphasis must be given to the training of surgeons in this new technique, so that major complications such as median nerve injuries can be avoided. In addition, future studies must better define the actual incidence of nerve injuries for both endoscopic carpal tunnel release and open carpal tunnel release in the community setting. PMID- 9734429 TI - Antibody to transforming growth factor beta reduces collagen production in injured peripheral nerve. AB - Epineurial scarring in peripheral nerve after injury inhibits normal axonal regeneration primarily due to fibroblast deposition of type I collagen. The transforming growth factor beta (TGF-beta) family is an important class of signaling molecules that has been shown to stimulate fibroblasts to produce collagen. The aim of this study was to design a prototypic therapeutic system in which the neutralization of TGF-beta in crushed rat sciatic nerve would decrease collagen formation. A total of 45 experimental Lewis rats were used. Group 1 animals (20 rats) sustained a unilateral crush injury to the sciatic nerve with injection of phosphate buffer solution. Group 2 animals (20 rats) sustained a unilateral crush injury to the sciatic nerve with injection of phosphate-buffered saline and goat, anti-rat, panspecific TGF-beta antibody. Group 3 control animals (five rats) underwent only exposure of sciatic nerve with injection of antibody. All animals were killed at 14 days and sciatic nerve specimens were harvested at that time. Slides of experimental tissue were processed using a 35S-labeled oligomer for procollagen alpha-1 mRNA, then dipped in photographic emulsion and examined by darkfield autoradiography. Morphometric analysis of pixel counts was then performed. A significant reduction in total pixel count per high-power field and in total number of fibroblasts per high-power field was found in crushed rat sciatic nerve treated with anti-TGF-beta antibody when compared with those treated only with phosphate-buffered saline. These findings are consistent with successful reduction in procollagen induction after a crush injury by topical administration of blocking antibody against transforming growth factor beta. The concept of growth factor blockade for therapeutic collagen reduction is attractive in the context of nerve injury, and the current article provides a model for future clinical application. PMID- 9734430 TI - Internal calvarial bone distraction in rabbits with delayed-onset coronal suture synostosis. AB - Recent studies have identified a subpopulation of craniosynostotic individuals who exhibit progressive or delayed-onset synostosis and mild craniofacial growth abnormalities. These individuals may be good candidates for nonextirpation, distraction osteogenesis therapy. The present study was designed to test this hypothesis by using internal calvarial bone distraction in a rabbit model with familial delayed-onset craniosynostosis. Data were collected from 159 rabbits: 71 normal controls, 72 with delayed-onset coronal suture synostosis, 8 with delayed onset coronal suture synostosis and coronal suturectomy, and 8 with delayed-onset coronal suture synostosis and distraction. At 10 days of age, all rabbits had amalgam markers placed on both sides of the frontonasal, coronal, and anterior lambdoidal sutures. At 25 days of age, correction was accomplished through either a 5-mm-wide suturectomy or distraction osteogenesis. An internal distraction appliance was fixed to the frontal and parietal bones and percutaneously and intermittently activated at an average of 0.10 mm/day for 42 days (4.11 mm total). Serial radiographs were taken at 10, 25, 42, and 84 days of age. Results revealed that rabbits with delayed-onset synostosis had significantly (p < 0.01) reduced coronal suture growth rates (0.04 mm/day) compared with the other three groups (0.07 mm/day). Rabbits with suturectomy and rabbits with distraction showed similar coronal suture responses. However, from 42 to 84 days of age, rabbits with distraction showed reduced growth at the vault sutures and abnormal growth patterns in cranial vault width, cranial vault shape, and cranial base angulation compared with the other three groups. Results demonstrated that, although the normal coronal suture growth rate was maintained in rabbits with delayed-onset synostosis using intermittent distraction osteogenesis, normal adult craniofacial structure was not achieved. Such anomalous growth was probably a result of altered growth vectors and compressive forces at adjacent sutures during distraction. These findings suggest that distraction osteogenesis without corticotomy may be a treatment alternative in individuals with progressive, delayed-onset synostosis, but that internal appliances that generate low-level, continuous distractive forces should be investigated and developed. PMID- 9734431 TI - Effect of lipectomy on growth and development of hyperinsulinemia and hyperlipidemia in the Zucker rat. AB - The Zucker fat rat inherits obesity and hyperinsulinemia, exhibits insulin resistance, and is, therefore, a model of adult onset, or type II, diabetes. The purpose of this study was to determine if excision of fat depots from the infant Zucker (fa+/fa+) rat would affect growth, fat cell number, hyperinsulinism, and hyperlipidemia. In the experimental design, 10 percent of the total body weight (inguinal and interscapular depots) was excised at 6 weeks of age from 18 fat and 18 lean (fa+/fa-) litter mates, with 18 fat and 18 lean rats serving as nonoperated controls. At intervals, serum glucose, insulin, cholesterol, and triglycerides were measured. Initially, the operated fat group was significantly (p < 0.01) lighter than the nonoperated group. By 9 weeks postoperatively, the operated fat rat group had regained weight and continued to grow at the same rate as the nonoperated fat rats because of intra-abdominal fat depots. Lipectomy had no effect on growth rate of the lean rat group. Although lipectomy caused no consistent change in serum glucose or insulin levels, it caused a significant decrease in lipid levels. For example, the operated fat rats had a reduction in cholesterol from 876 to 171 mg/dl by 15 weeks postoperatively, and serum cholesterol persisted at about 50 percent of the nonoperated group throughout the rest of the study (38 weeks postoperatively). Even a greater reduction in triglyceride levels occurred, for example, from 7415 to 1082 mg/dl at 24 weeks postoperatively. Lipectomy did not cause a change in lipid levels in the lean group. It is concluded that the lipectomy in the Zucker fat group is an excellent model to evaluate the effects of changes in fat cell number on lipid metabolism. PMID- 9734432 TI - A microangiographic technique using synchrotron radiation to visualize dermal circulation in vivo. AB - Conventional angiography cannot resolve dermal small vessels with a diameter of 200 microm or less. In vitro microangiography is currently characterized by better spatial resolution than conventional angiography but does not allow visualization of the blood stream in dermal vessels in vivo. In this study, we introduce a novel synchrotron radiation microangiographic system for visualizing the structure of and blood flow in dermal microvessels in vivo repeatedly. We used monochromatic synchrotron radiation with an energy just above the k-edge of iodine (33.3 keV) as an x-ray and a high-definition television camera system with a high-sensitivity image pick-up tube for detection. The 33.3-keV monochromatic synchrotron radiation allows detection of a small amount of iodine, and the high definition television camera system can resolve small vessels with high-spatial resolution and no loss of sensitivity. We performed synchrotron radiation angiography of superficial inferior epigastric arteries and their branches in nine rats, and of the caudal artery in 14 rats, and compared angiographic images taken by the current system with those taken by a conventional angiographic system in seven rats. With this new microangiographic technique, we could visualize small dermal vessels with a diameter as low as 50 microm. In addition, repeated angiograms at baseline and under increasing body temperature could be obtained. This new microangiographic approach is expected to be very useful for the assessment of dermal circulation in patients. PMID- 9734433 TI - The effects of surgical and chemical delay procedures on the survival of arterialized venous flaps in rabbits. AB - The aim of this study was to investigate the efficacy of a surgical delay procedure and a combined surgical and chemical delay procedure on the survival of arterialized venous flaps. Experimental groups included (1) a control group, (2) a surgical delay (4-day and 7-day delay) group, and (3) a combined surgical and chemical (doxazosin mesylate, nitroglycerine patch) delay group. These groups were further divided into subgroups (n = 10) depending on the delay period and the chemical agents. An arterialized venous flap was created on one ear of each rabbit. In the arterialized venous flap, arterial inflow was provided by anastomosis of the central auricular artery to the anterior branch of the central auricular vein and a venous outflow through the anterior marginal vein. In the control group, the arterialized venous flaps without any delay procedure showed complete necrosis of all flaps. In the surgical delay group, the mean percentage survival of arterialized venous flaps was 36.6 percent in the 4-day delay group and 59.7 percent in the 7-day delay group. In the combined surgical and chemical delay group, a 3-day chemical delay followed by a 4-day simultaneous surgical and chemical delay resulted in mean percentage survival of the arterialized venous flaps of 81.1 percent in the doxazosin mesylate group, 72.8 percent in the nitroglycerine patch group, and 92.9 percent in a combination group of doxazosin mesylate and nitroglycerine patch. A 3-day chemical delay followed by a 7-day simultaneous surgical and chemical delay resulted in mean percentage survival of the arterialized venous flaps of 94 percent in the doxazosin mesylate group, 90.2 percent in the nitroglycerine patch group, and 99 percent in a combination group of doxazosin mesylate and nitroglycerine patch. In conclusion, the surgical delay procedure increased the percentage survival of the arterialized venous flaps in proportion to the delay period. The combination group of surgical and chemical delay procedures had a significantly greater percentage survival than that of the surgical delay group (p < 0.001), and the delay period could be shortened. PMID- 9734434 TI - Cranioplasty in frontometaphyseal dysplasia. AB - Frontometaphyseal dysplasia is an extremely rare craniotubular bone disorder predominantly manifested by supraorbital bossing. Although recognizable with findings at birth, it is usually identified successfully before the onset of puberty. These patients often are stigmatized related to their appearance and may present to the plastic surgeon for intervention. We present a case of successful cranioplasty in correcting the fronto-orbital deformity in a 9-year-old child with frontometaphyseal dysplasia. PMID- 9734435 TI - Facial reconstruction using a retroauricular-temporal free flap. AB - Large and deep soft-tissue defects of the face usually require resurfacing by free-tissue transfer. An appropriate free flap for facial reconstruction may be harvested from the retroauricular and temporal region utilizing two arterial pedicles (superficial temporal artery and posterior auricular artery). This flap provides normal color, texture, and thickness and thus is an optimal anatomic and aesthetic reconstruction with minimal donor-site morbidity. PMID- 9734436 TI - Feminizing Sertoli cell tumors associated with Peutz-Jeghers syndrome: an increasingly recognized cause of prepubertal gynecomastia. AB - Testicular sex cord tumors with annular tubules are an increasingly recognized cause of prepubertal gynecomastia typically accompanied by accelerated linear growth and advanced bone maturation. Serum estrogen levels may be elevated. Testicular ultrasound and biopsy are diagnostic, and mastectomy is indicated. Although these tumors can occur independently, causing gynecomastia in 10 percent of cases, they usually occur in patients with Peutz-Jeghers syndrome. In any Peutz-Jeghers syndrome patient developing gynecomastia, a testicular tumor should be sought. Conversely, because a significant proportion of all reported prepubertal gynecomastia patients have Peutz-Jeghers syndrome with testicular tumors, this syndrome must be considered for all young boys in whom the cause of gynecomastia is not otherwise apparent. When Peutz-Jeghers syndrome is suspected, gastroscopy, colonoscopy, and testicular biopsies can be performed under one anesthetic at the time of mastectomy. PMID- 9734437 TI - Congenital fibrosarcoma of the upper extremity. AB - Congenital or infantile fibrosarcoma is a rare soft-tissue neoplasm that should be considered in the differential diagnosis of a large extremity mass presenting at birth. These tumors are notoriously misdiagnosed at birth as either hemangiomas or lymphatic malformations. Definitive diagnosis is made by physical examination, special radiologic studies, and biopsy. Although histologically similar to fibrosarcomas occurring in adults, the congenital lesions differ in their clinical behavior; metastases are rare, local recurrence is common, and the prognosis is good with wide local excision combined with chemotherapy. Amputation should be reserved for chemoresistant patients in whom the involvement of neurovascular structures by the tumor make a limb-sparing aggressive excision impossible. PMID- 9734438 TI - Neurotmesis of the lateral femoral cutaneous nerve when coring for iliac crest bone grafts. AB - Two patients are presented suffering from meralgia paresthetica, owing to neurotmesis of the lateral femoral cutaneous nerve following bone-graft harvesting from the iliac crest with a cylindrical osteotome. Despite this donor site complication, the advantages of a coring technique as compared with traditional bone-graft harvesting techniques are numerous: better cosmetic results, less pain, no need for general anesthesia, shorter hospitalization, and a minimum of donor-site morbidity. Also, this technique has proven itself easy and effective. To minimize donor-site morbidity, some clinical recommendations to avoid neurotmesis are given with regard to the anatomy and anatomic variations of the lateral femoral cutaneous nerve. PMID- 9734439 TI - A new twist to the myocutaneous turnover flap for closure of a spinal defect. AB - We have described the deepithelialized myocutaneous turnover of a gluteus maximus island flap based on the superior gluteal vessels used for closure of an acquired midline defect of the lower back complicated by cerebrospinal fluid fistula. Cerebrospinal fluid fistula, although a rare complication of an acquired midline back defect, further adds to the reconstructive challenge of such a wound. As this case illustrates, cerebrospinal fluid fistula can have serious and potentially life-threatening implications and so demands rapid surgical intervention. Dural patching with fascia lata graft having already failed, reconstruction was achieved using a well vascularized deepithelialized cutaneous patch on a gluteus maximus island turnover flap. The technique of deepithelializing a skin island for dural repair could be applied to any myocutaneous flap used in reconstruction of a midline back defect complicated by cerebrospinal fluid leak. This provides a method of dural repair and reconstruction of the defect in one step and obviates the need for cerebrospinal fluid diversion. PMID- 9734440 TI - A new operative method for treatment of xanthelasma or xanthoma palpebrarum: microsurgical inverted peeling. AB - A new operative technique for treatment of xanthelasma or xanthoma palpebrarum was introduced. The merits of this technique are (1) no skin excision with the tumor, (2) making the flap from the skin covering the tumor, (3) no need for donor site, (4) no significant cosmetic problem, and (5) reoperation performed by same procedure. Our new technique is performed under local anesthesia. The skin and tumor are raised together just above the orbicularis oculi muscle like a flap. The tumor is peeled piece by piece from the flap using micro-scissors under a surgical microscope. After tumor enucleation is performed, the skin flap without the tumor is sutured back directly. Our new technique was performed for seven cases of xanthelasma. The recurrence of the tumor was seen only in two cases, which were complicated with hypercholesterolemia. No recurrence was observed after a secondary operation and with the use of anti hypercholesterolemic drugs. Large xanthelasma can be treated by our new surgical technique without extra skin excision with the best possible cosmetic results. PMID- 9734441 TI - A single-stage two-flap method for reconstruction of partial auricular defect. AB - Two flaps, consisting of a postauricular skin flap and a mastoid fascial flap, were used for coverage of both sides of a grafted framework in one stage for repair of one-fourth to half auricular composite defect after trauma or inflammation. The method was safe, and the postauricular donor scar was minimal. Additionally, it was beneficial in restoring similar ear shape and size with a natural cartilage convolution. PMID- 9734442 TI - Integration of the central mound technique with the vertical skin takeout reduction mammaplasty. PMID- 9734443 TI - Preexpansion of the tensor fasciae latae for free-flap transfer. AB - Preexpansion has become an established technique to prefabricate elective free flap transfers. We report the use of the tensor fasciae latae flap as a donor site in two cases for reconstruction of a burn scar neck contracture and an unstable contralateral below-knee amputation stump, of which other donor sites were ruled out either by the patients' condition or by choice. Implantation and transfer were straightforward and the donor sites of very large flaps were minimized by preexpansion. The preexpanded muscle fasciocutaneous flaps were transplanted with microsurgical anastomoses of the vessels. Apart from a small area of necrosis at the distal tip of the flap developing on the sixth postoperative day, which we excised in a second operation, there were no major complications. The advantages of the combination of preexpansion and free flap transfer as well as the unique anatomical and functional qualities of this musculocutaneous unit are emphasized. PMID- 9734444 TI - Simple technique of skin grafting with a single-use safety razor. AB - Harvesting a split-thickness skin graft by a single-use safety razor is described. PMID- 9734445 TI - A new and reliable method of securing skin grafts to the difficult recipient bed. PMID- 9734446 TI - Endoscopic application in aesthetic and reconstructive facial bone surgery. AB - Twenty-three cases of endoscopically assisted facial bone surgery were performed over the past 3 years. Our series is consistent with 16 cases of aesthetic contouring surgery and 12 treatments of facial bone fracture, including three cases for recontouring of frontal bone, three cases for recontouring of zygoma, endoscopically assisted correction of three zygomatic and blowout fractures, four cases for rhinoplasty and septoplasty for deviated nose, and three cases for mandible contouring surgery. To accomplish this technique, a rigid 4-mm, 30 degree down-angled endoscope was used. The frontal bone or zygomatic arch was approached endoscopically through two or three small incisions on the frontal or temporoparietal scalp. All endoscopic instruments were then manipulated through these incisions. The approach for endoscopically assisted rhinoplasty is the same as with standard rhinoplasty procedures. The approach for zygoma complex and maxillary sinus needs an intraoral incision. Recontouring of zygoma, mandible, and nasal dorsum by an air-driven burr and rasp was performed with endoscopic visual assistance. A plate and screw fixation for zygomatic arch fracture requires an additional small skin incision over the plate for the trocar method. The duration of follow-up ranged from 6 months to 30 months. The postoperative course was satisfactory with a few complications. The extra time needed for the endoscopic procedures was less than 1 hour. Endoscopically assisted facial bone surgery can be performed with adequate visualization and direct manipulation of all facial bones. Complications usually associated with extensive incisions in the bicoronal approach may be avoided. Poor visualization in the conventional approach for operation of orbit, nose, maxillae, and mandible may be avoided by use of the endoscope. This technique may prove to be ideal for aesthetic surgery for facial skeleton with smaller scar and less morbidity. PMID- 9734447 TI - Characterization of portwine stain disfigurement. AB - Portwine stain disfigurement is caused by several factors. To what extent and in which proportion these factors influence the overall perceived disfigurement is incompletely understood. In this study, the contribution of seven portwine stain characteristics to overall portwine stain disfigurement was assessed. Color slides were taken from 90 patients with untreated portwine stains in the head/neck area. From these slides, overall portwine stain disfigurement was judged by a panel of 16 lay persons. The reliability of the average ratings of this panel was established with weighted kappa analysis (kappa = 0.51) and by calculating the Cronbach alpha coefficient (0.99). Using a previously tested multi-item questionnaire, the following portwine stain characteristics were rated quantitatively by a panel of five professionals: color, patchiness, boundary, size, shape, surface structure, and hypertrophy of the underlying tissue. By means of multiple linear regression analysis, the ratings for overall portwine stain disfigurement (panel of lay persons) were compared with the ratings for the individual portwine stain characteristics (panel of professionals). From the results of this analysis, the percentual contribution of each of the characteristics to overall portwine stain disfigurement was calculated. Size turned out to be the most important portwine stain characteristic, being responsible for almost half of the overall disfigurement. Color and boundary are the next two most important characteristics, contributing 18.7 and 12.4 percent, respectively. The other four characteristics together account for 10 percent. In our model, 13 percent of overall portwine stain disfigurement remains unexplained. We expect patient features to account for this. We feel that these results may have consequences for laser treatment of portwine stains. Reducing the size and fading out the boundary of the stain probably reduce overall portwine stain disfigurement more effectively than primarily trying to lighten the often persistent center of the stain. PMID- 9734448 TI - Diplopia following transconjunctival blepharoplasty. AB - The resurgence of popularity of the transconjunctival approach to lower eyelid fat removal as a component of cosmetic blepharoplasty has been highlighted by a number of publications in recent years. There has been, however, minimal discussion in the literature of the complications of this procedure. Although the mechanism of muscle injury is similar in transcutaneous and transconjunctival surgery, there is a much more direct route to the inferior extraocular musculature via the latter approach. Herein, we present a series of six patients with diplopia status post-transconjunctival lower eyelid blepharoplasty referred to the Manhattan Eye, Ear, and Throat Hospital for evaluation. Transconjunctival lower lid blepharoplasty was performed as a primary procedure in four patients and as a secondary procedure following transcutaneous blepharoplasty in two patients. Patients were evaluated with ocular examination and orthoptic measurements. Magnetic resonance imaging was obtained in two cases. The inferior rectus and inferior oblique muscles were found to be equally injured in these cases (4 of 6), and the lateral rectus was encountered in one case. Two patients required strabismus surgery to correct their diplopia, whereas four patients improved with observation alone. The possible etiologies of postoperative diplopia following transconjunctival lower lid blepharoplasty are manifold. Mechanisms of extraocular muscle injury may include intramuscular hemorrhage and edema, cicatricial changes within the muscle, and accidental incorporation of extraocular muscle in closure of orbital septum. Avoidance of these complications is probably best achieved through intimate understanding on the part of the surgeon of eyelid anatomy from the transconjunctival perspective. PMID- 9734449 TI - Open rhinoplasty: columellar scar analysis in an Arabian population. AB - This prospective study included 50 consecutive Saudi Arabian patients who underwent open rhinoplasty. All patients had a minimum follow-up of 1 year after surgery. The columellar scar was assessed objectively and subjectively at the final follow-up. An unsatisfactory scar was considered present if anything other than a barely visible, level, and thin-line scar without notching was evident. Objectively, 39 patients (78 percent) were considered to have a satisfactory columellar scar. The percentage of unsatisfactory scars (22 percent) was higher than expected prior to performing the study, and one patient was actually conscious about the deformity. Causes and prevention of unsatisfactory columellar scars were discussed. PMID- 9734450 TI - Silicone in nasal augmentation rhinoplasty: a decade of clinical experience. AB - A number of materials, both biologic and alloplastic, have been used for nasal augmentation. Although biologic bone and cartilage grafts are associated with lower infection rates, they are also associated with long-term resorption and donor-site morbidity. Alloplastic materials, in particular silicone, have been associated in the literature with extrusion and infection but have the advantages of being affordable and easy to reshape with no requirement for harvesting autografts. A 10-year experience with silicone nasal augmentation documenting clinical experience, acute and long-term complications, and patient satisfaction was reviewed. All patients undergoing silicone augmentation rhinoplasty between July of 1985 and December of 1995 were reviewed. Preoperative nasal phenotype, operative data, and postoperative outcome were recorded. Long-term follow-up was undertaken using a telephone survey. There were 422 patients who underwent silicone nasal augmentation from July of 1985 to December of 1995. Only nine were men. The indications were for aesthetic nasal augmentation in 98 percent, and the majority (98 percent) were of South East Asian origin. Mean age was 26 (range 17 to 36), and 41 of the 422 patients had had previous nasal augmentation performed before presentation. Twenty-three patients (5.5 percent) had complications requiring removal of the implant within 30 days of surgery. These included displacement, prominence, hemorrhage, and excessive pressure in addition to obvious supratip deformity. On late follow-up, a further 18 patients (4.3 percent) had subsequent removal of the prosthesis. The most common reason for this was either displacement or over-prominence, more often judged by the surgeon than the patient. There were only two patients (0.5 percent) who had extrusion of the prosthesis. A total of 266 patients (63 percent) were contacted for a telephone interview. The majority of patients (84.2 percent) were satisfied with their nasal shape. Of the 42 patients (15.8 percent) who were not satisfied, 21 patients still wanted further augmentation of their nose. Photographic analysis of 198 patients showed a mean augmentation of 16.5 percent (range 4.0 to 27.5). Amount of augmentation correlated with preoperative nasal phenotype. Silicone nasal augmentation is a safe and effective procedure when used for moderate increases in nasal height. Contrary to previous reports, this series showed no associated infection. If the implant is shaped appropriately to the patient's nasal phenotype, the risk of extrusion may be reduced. PMID- 9734451 TI - Combined chemical peeling and dermabrasion for deep acne and posttraumatic scars as well as aging face. AB - The combination of chemical peeling and dermabrasion for the improvement of facial wrinkles, acne, posttraumatic scars, and abnormal pigmentation was first described by Dupont in 1972 and Horton in 1984. We have been using the combined technique since 1972, and we have obtained more satisfying results than by using these techniques independently. The purpose of this paper is to summarize the results obtained using the combined technique of chemical peeling and dermabrasion and to emphasize a simple method of postoperative care that can be applied after any physical or chemical rejuvenation technique. Whereas the combined technique takes advantage of depth-controlled surgery, less bleeding, less postoperative pain, less risk of local and systemic complications, and longer lasting results, the covering of the wound with one layer of fine mesh gauze is another advantage that provides easy postoperative care. PMID- 9734453 TI - Correction of inverted nipple using strut reinforcement with deepithelialized triangular flaps. PMID- 9734452 TI - Thermal injuries as a result of CO2 laser resurfacing. AB - CO2 laser resurfacing of the face for fine wrinkles has gained great popularity over a short period of time. The use of the CO2 laser has proven to be effective in reducing or eliminating fine wrinkles. This tool in the surgeon's armamentarium has been added to those of dermabrasion and chemical peel. The theoretical advantage of the use of the CO2 laser for resurfacing has been better accuracy and reportedly more control of the depth of penetration. The use of the CO2 laser has been welcomed by many cosmetic surgeons. Until now, there have been few reported cases of complications with the use of the CO2 laser. To many, this would sound too good to be true; unfortunately, that is the case. The CO2 laser is a high-energy machine that can indeed cause thermal injury. This thermal injury can result in deep burns to the skin and hypertrophic scarring. We feel this is more common than is currently being reported, and we share our experience as a burn and wound care referral service. During an 18-month period, 20 consecutive patients were referred to our practice who had received injuries from the CO2 laser resurfacing laser. We present here in this review a summary of those injuries. The CO2 resurfacing laser is a very effective tool for the treatment of fine wrinkles, but it is not without the potential for serious complications. We urge caution with the use of the laser and prompt recognition and treatment of thermal injury to the skin. PMID- 9734454 TI - A new method for determining bra size and predicting postaugmentation breast size. AB - The current popular system of determining bra size is inaccurate so often as to be useless. Add to this the many different styles of bras and the lack of standardization between brands, and one can see why finding a comfortable, well fitting bra is more a matter of educated guesswork, trial, and error than of precise measurements. An improved method of determining cup size by directly measuring the circumference of the breast is presented in this paper. If adopted commercially, this simple method could reduce dramatically the number of women said to be wearing the "wrong-sized bra." The technique has also been found to be an accurate predictor of postoperative breast size in augmentation and reduction surgery, and thus offers plastic surgeons an opportunity to better meet the expectations of women undergoing these procedures. PMID- 9734456 TI - The new Medicare regulations: a very hidden opportunity. PMID- 9734455 TI - Cruel and excessive punishment. PMID- 9734457 TI - Anchors aweigh. Plastic Surgery Educational Foundation DATA Committee. Device and Technique Assessment. PMID- 9734458 TI - Retin-A Micro. Plastic Surgery Educational Foundation DATA Committee. Device and Technique Assessment. PMID- 9734459 TI - Plastic surgery of the face in Byzantium in the fourth century. AB - Oribasius was an eminent Byzantine physician who lived in the fourth century. His greatest contribution to medical history was his anthology of all important medical works of his time, entitled Synagogue Medicae. This complete medical encyclopedia of his era consisted of more than 70 volumes. A significant part of this work has been lost. What remains, however, allows us to glimpse the surprising richness and knowledge of ancient medicine. Chapters 25 and 26 of the original 42nd book of Oribasius are of specific interest to the plastic surgeon, because they deal with reconstruction of facial defects. Reconstructive procedures for defects in the eyebrows, forehead, cheeks, nose, and ears are described. Advancement flaps are suggested for the reconstruction, and recommendations are made about debridement, flap design, and thickness of flaps. It becomes obvious from these texts that Greek, Roman, and Byzantine surgeons had the knowledge and experience to perform several reconstructive procedures of the face and nose. This knowledge was passed to the Arabs and then to Western Europe in the 15th century and became part of the foundation for modern plastic surgery. PMID- 9734460 TI - Imaging signs and radiologists' jargon of ruptured breast implants. AB - Silicone gel leakage problems are central to the furor over the complications alleged to be caused by breast implants. Because clinical examination may not reveal confirmatory signs of gel bleed or rupture, radiologists are often requested by plastic surgeons to evaluate the integrity of the implant's envelope. The findings of the various imaging investigations are reported in terms such as "teardrop," "linguini," and "snowstorm." To interpret the radiologist's report correctly, the plastic surgeon should be familiar with these terms and the findings they represent. In this article, we present an explanation of the radiologists' vocabulary in these matters, as well as an indication as to the significance of the various signs. PMID- 9734461 TI - What age for face lifts? PMID- 9734462 TI - Topical tretinoin and smokers' face lifts. PMID- 9734463 TI - Suitability of minimally invasive iliac cancellous bone harvesting on children. PMID- 9734464 TI - Determining the frequency of breast implant failure requires sound scientific principles. PMID- 9734465 TI - The TRAM delay: burning a lifeboat? PMID- 9734466 TI - Turbocharge or supercharge? PMID- 9734467 TI - An update on endoscopic sural nerve harvest. PMID- 9734468 TI - Safari surgery. PMID- 9734469 TI - Troponin and tropomyosin: proteins that switch on and tune in the activity of cardiac myofilaments. AB - We present a current perception of the regulation of activation of cardiac myofilaments with emphasis on troponin (Tn) and tropomyosin (Tm). Activation involves both a Ca2+-regulated molecular switch and a potentiated state, dependent on feedback effects of force-generating crossbridges. Recent developments in the elucidation of the structure and arrangement of the myofilament proteins offer insights into the molecular interactions that constitute the switching and potentiating mechanisms. Transgenic mice overexpressing myofilament proteins, in vitro studies of mutant myofilament proteins, multidimensional multinuclear nuclear magnetic resonance, and fluorescence resonance energy transfer offer important approaches to understanding the molecular signaling processes. These studies reveal special features of the cardiac myofilament proteins that appear specialized for the unique functions of the heart. An important aspect of these special features is their role in mechanical, chemical, and neurohumoral coupling processes that tune myofilament activation to hemodynamics and beating frequency. Understanding these processes has become essential to understanding cardiac pathologies such as heart failure, ischemia and reperfusion injury, stunning, and familial hypertrophic cardiac myopathies. PMID- 9734470 TI - Microtubule involvement in translational regulation of fibronectin expression by light chain 3 of microtubule-associated protein 1 in vascular smooth muscle cells. AB - Our previous studies suggested that enhanced fibronectin mRNA translation in ductus arteriosus compared with aortic smooth muscle cells is related to increased expression of light chain 3 (LC3) of microtubule-associated protein 1, which binds an AU-rich element in the 3' untranslated region of fibronectin mRNA. We therefore hypothesized that microtubules are involved in LC3-mediated fibronectin mRNA translational regulation. In this study we show that disruption of microtubules by colchicine inhibits fibronectin mRNA translation in cultured ductus arteriosus smooth muscle cells. We proposed that the mechanism might be related to decreased docking of fibronectin mRNA on the translational machinery, ie, membrane-bound polysomes on rough endoplasmic reticulum, and confirmed this by Northern blot analysis. To investigate the mechanism further, we carried out polysome analysis using sucrose gradient centrifugation and fractionation and studied the polysomal distribution of fibronectin mRNA and LC3 protein in the sucrose gradient by using RNase protection assay and Western immunoblotting, respectively. Colchicine treatment shifts fibronectin mRNA from the fractions containing membrane-bound polysomes to the fractions carrying free polysomes and concomitantly decreases the amount of LC3 protein in the fractions containing membrane-bound polysomes. Furthermore, an EDTA-release experiment demonstrates that LC3 protein associates with the 60S ribosomal subunit. Our data support the concept that microtubules may function with LC3 to facilitate sorting of fibronectin mRNA onto rough endoplasmic reticulum and translation. PMID- 9734471 TI - 3-Hydroxy-3-methylglutaryl coenzyme a reductase and isoprenylation inhibitors induce apoptosis of vascular smooth muscle cells in culture. AB - Recent evidence suggests that apoptosis may be involved in the control of vascular smooth muscle cell (VSMC) number in atherosclerotic lesions. 3-Hydroxy-3 methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors have been reported to induce apoptosis in a variety of tumor cell lines. To evaluate whether these agents also induce apoptosis of VSMCs, cultured rat VSMCs were treated with increasing doses of atorvastatin in the presence of FBS as a survival factor. The presence of apoptosis was evaluated by morphological criteria, annexin V binding, and DNA fragmentation and quantified as the proportion of hypodiploid cells by flow cytometry. Atorvastatin induced apoptosis in a dose-dependent manner, an effect also seen with simvastatin and lovastatin, but not with the hydrophilic drug pravastatin. The proapoptotic effect of statins was seen only when the inhibition of acetate incorporation into sterols was >95% and was fully reversed by mevalonate, farnesyl pyrophosphate, and geranylgeranyl pyrophosphate but not by isopentenyl adenosine, ubiquinone, or squalene, suggesting a role for prenylated proteins in the regulation of VSMC apoptosis. To further assess the role of protein prenylation, VSMCs were exposed to the prenyl transferase inhibitors perillic acid and manumycin A. Both agents induced VSMC apoptosis as evaluated by the above-mentioned criteria. Finally, VSMC treatment with lipophilic statins was associated with decreased prenylation of p21-Rho B, further supporting the role of protein prenylation inhibition in statin-induced VSMC apoptosis. The present data suggest that interference with protein prenylation by HMG-CoA reductase inhibitors or other agents may provide new strategies for the prevention of neointimal thickening. PMID- 9734472 TI - Human coronary arteriolar dilation to arachidonic acid depends on cytochrome P 450 monooxygenase and Ca2+-activated K+ channels. AB - Endothelium-dependent hyperpolarization of vascular smooth muscle cells (VSMCs) plays a crucial role in regulating vascular tone, especially in resistance vessels. It has been proposed that metabolites of arachidonic acid (AA), formed by cytochrome P-450 monooxygenase (P450), are endothelium-derived hyperpolarizing factors (EDHFs). These metabolites have been reported to mediate dilation to endogenous vasoactive compounds, such as bradykinin and acetylcholine. However, it is not known whether these metabolites of AA contribute to dilation of human resistance vessels. This is important since it has been proposed that EDHF serves as a compensatory mechanism to maintain dilation in disease states. Therefore, we studied the effect of AA on vessel diameter and VSMC membrane potential in isolated human coronary microvessels. Arterioles (81+/-5 microm, n=70) were dissected from right atrial appendages at the time of cardiac surgery and cannulated at a distending pressure of 60 mm Hg and zero flow. Changes in internal diameter were recorded with videomicroscopy. Some vessels were impaled with glass microelectrodes to measure membrane potential of VSMCs while internal diameters were simultaneously recorded. After constriction (47+/-2%) with endothelin-1, AA (10(-10)to 10(-5)mol/L) induced substantial dilation of human coronary microvessels, which was abolished by removal of the endothelium. Treatment with 17-octadecynoic acid (17-ODYA, 10(-5) mol/L; a P450 inhibitor) attenuated maximal dilation to AA (49+/-9% versus 91+/-4% [control]; P<0.05 versus control), whereas indomethacin (INDO, 10(-5) mol/L; a cyclooxygenase inhibitor) and N omega-nitro-L-arginine methyl ester (L-NAME, 10(-4) mol/L; a NO synthase inhibitor) were without effect. Both 17-ODYA and miconazole (10(-5) mol/L, a chemically distinct P450 inhibitor) further reduced the dilation to AA in the presence of INDO. The presence of 40 mmol/L KCl or charybdotoxin (10(-8) mol/L, a blocker of large-conductance Ca2+-activated K+ channels) impaired dilation to AA (19+/-9% [KCI] versus 76+/-5% [control] and 47+/-6% [charybdotoxin] versus 91+/-3% [control]; P<0.05 for both). After depolarization with endothelin-1 (-26+/-1 mV from -48+/-3 mV [before endothelin]), AA (10( 5)mol/L) in the presence of INDO and L-NAME induced hyperpolarization of VSMCs ( 57+/-5 mV). In the presence of 17-ODYA together with INDO and L-NAME, endothelin produced similar depolarization (-26+/-2 mV from - 48+/- 3 mV), but hyperpolarization to AA was reduced (-33+/-2 mV; P<0.05 versus absence of 17 ODYA). AA metabolites formed primarily by P450 produce potent endothelium dependent dilation of human coronary arterioles via opening of Ca2+-activated K+ channels and hyperpolarization of VSMCs. These findings support an important role for P450 metabolites in the regulation of human coronary arteriolar tone. PMID- 9734473 TI - Lysophosphatidylcholine enhances cytokine-induced interferon gamma expression in human T lymphocytes. AB - Accumulation of substantial numbers of activated T lymphocytes, as well as monocyte/macrophages, in focal areas of arterial intima appears to be a hallmark of atherogenesis. Our previous report demonstrated that lysophosphatidylcholine (lyso-PC), a polar phospholipid component that is increased in atherogenic lipoproteins and atherosclerotic lesions, can upregulate the expression of heparin-binding epidermal growth factor-like growth factor and the interleukin (IL)-2 receptor in cultured human peripheral T lymphocytes. In this study, we show that lyso-PC can also enhance interferon gamma (IFN-gamma) secretion and gene expression in human T lymphocytes. Lyso-PC-induced upregulation of IFN-gamma depended on the presence of IL-2, IL-12, or phytohemagglutinin in culture media and was similarly observed in both CD4+ and CD8+ subsets. Actinomycin D chase by Northern blotting showed that lyso-PC significantly prolonged IFN-gamma mRNA half lives in human T cells. Transient transfection of IFN-gamma promoter-reporter gene construct in the human T-cell line Jurkat cells demonstrated that lyso-PC stimulated the transcription of IFN-gamma promoter-driven luciferase gene. Analyses of serial deletion mutations of IFN-gamma promoter revealed that the lyso-PC-responsive element is located between base pairs - 102 and -78 of the transcription initiation site of the IFN-gamma gene. Enhanced expression of IFN gamma in T lymphocytes by lyso-PC may play a crucial role in atherogenesis. PMID- 9734474 TI - Regulation of cardiomyocyte apoptotic signaling by insulin-like growth factor I. AB - Apoptosis is regulated by specific intracellular signaling pathways. The development of cardiomyopathy involves the apoptosis of cardiomyocytes; however, the details of their apoptotic signaling are not yet known. Insulin-like growth factor I (IGF I) is an important survival growth factor for myocardium and other tissues, but the effects of IGF I on apoptotic signaling remain largely unknown. To study apoptotic signaling pathways in cardiomyocytes and to understand IGF I actions on the apoptotic signaling of cardiac muscle cells, we have defined the effects of IGF I on Bcl-2, Bax, caspase 3, DNA fragmentation, and cell survival in primary cardiomyocytes. Compared with Bax levels, the levels of Bcl-2 were found to be quite low in these cells. Serum withdrawal and doxorubicin reduced cell viability, increased fragmentation of DNA, increased cellular contents of Bax, and activated caspase 3. IGF I enhanced cell viability, suppressed DNA fragmentation, attenuated Bax induction, and suppressed caspase 3 activation. The levels of Bcl-2-associated Bax were increased after serum withdrawal and incubation with doxorubicin and were reduced by IGF I. Thus, cardiomyocyte apoptosis induced by serum withdrawal and doxorubicin likely results, in part, from the induction of Bax and activation of caspase 3, but IGF I may inhibit cardiomyocyte apoptosis by attenuating Bax induction and caspase 3 activation. These findings provide new insight into the mechanisms of cardiomyocytes apoptosis and may help elucidate how IGF I modulates apoptotic signaling in cardiac muscle. PMID- 9734475 TI - Versican expression is associated with chamber specification, septation, and valvulogenesis in the developing mouse heart. AB - The versican (PG-M) gene encodes a chondroitin sulfate proteoglycan that is nonpermissive for cell migration and appears in association with slow cell proliferation and cytodifferentiation. Using the techniques of in situ hybridization and immunocytochemistry on sectioned mouse embryos, we found that the mRNA and protein for versican show similar distributions and are expressed in a dynamic pattern during development of the heart. Versican exhibits generalized expression in the tubular heart but becomes rapidly downregulated in the atrium and exhibits higher transcript levels on the right side of the ventricular chamber than the left, before the onset of ventricular septation. Versican is expressed strongly in the trabeculated ventricular myocardium, whereas the compact proliferative zone has lower transcript abundance. It is expressed in the outer layers and on the crest of the ventricular septum and is prominent on the mesenchymal cap of the primary atrial septum. Versican is particularly strongly expressed in the endocardial cushions of the atrioventricular and outflow tract regions and in the atrioventricular, semilunar, and venous valves. This study raises the possibility that versican may be involved in specification of the ventricular chambers, in growth and fusion of the atrial and ventricular septa, and in the transformation from epithelium to mesenchyme that characterizes development of the endocardial cushions. Versican may be a key participant in cardiogenesis, responding to the many diffusible signals that mediate interactions between the developing endocardium and myocardium. PMID- 9734476 TI - Plasmalogen-derived lysolipid induces a depolarizing cation current in rabbit ventricular myocytes. AB - Plasmalogen rather than diacyl phospholipids are the preferred substrate for the cardiac phospholipase A2 (PLA2) isoform activated during ischemia. The diacyl metabolite, lysophosphatidylcholine, is arrhythmogenic, but the effects of the plasmalogen metabolite, lysoplasmenylcholine (LPLC), are essentially unknown. We found that 2.5 and 5 micromol/L LPLC induced spontaneous contractions of intact isolated rabbit ventricular myocytes (median times, 27.4 and 16.4 minutes, respectively) significantly faster than lysophosphatidylcholine (>60 and 37.8 minutes, respectively). Whole-cell recordings revealed that LPLC depolarized the resting membrane potential from -83.5+/-0.2 to -21.5+/-1.0 mV. Depolarization was due to a guanidinium toxin-insensitive Na+ influx. The LPLC-induced current reversed at -18.5+/-0.9 mV and was shifted 26.7+/-4.2 mV negative by a 10-fold reduction of bath Na+ (Na+/K+ permeability ratio, approximately 0.12+/-0.06). In contrast, block of Ca2+ channels with Cd2+ and reducing bath Cl failed to affect the current. The actions of LPLC were opposed by lanthanides. Gd3+ and La3+ were equally effective inhibitors of the LPLC-induced current and equally delayed the onset of spontaneous contractions. However, the characteristics of lanthanide block imply that Gd3+-sensitive, poorly selective, stretch-activated channels were not involved. Instead, the data are consistent with the view that lanthanides increase phospholipid ordering and may thereby oppose membrane perturbations caused by LPLC. Plasmalogens constitute a significant fraction of cardiac sarcolemmal choline phospholipids. In light of their subclass-specific catabolism by phospholipase A2 and the present results, it is suggested that LPLC accumulation may contribute to ventricular dysrhythmias during ischemia. PMID- 9734477 TI - Ionic mechanisms of regional action potential heterogeneity in the canine right atrium. AB - Atrial action potential heterogeneity is a major determinant of atrial reentrant arrhythmias, but the underlying ionic mechanisms are poorly understood. To evaluate the basis of spatial heterogeneity in canine right atrial repolarization, we isolated cells from 4 regions: the crista terminalis (CT), appendage (APG), atrioventricular ring (AVR) area, and pectinate muscles. Systematic action potential (AP) differences were noted: CT cells had a "spike and-dome" morphology and the longest AP duration (APD; value to 95% repolarization at 1 Hz, 270+/-10 ms [mean+/-SEM]); APG and pectinate muscle cells had intermediate APDs (180+/-3 and 190+/-3 ms, respectively; P<0.001 versus CT for each), with APG cells having a small phase 1; and AVR cells had the shortest APD (160+/-4 ms, P<0.001 versus other regions). The inward rectifier and the slow and ultrarapid delayed rectifier currents were similar in all regions. The transient outward K+ current was significantly smaller in APG cells, explaining their small phase 1 and high plateau. L-type Ca2+ current was greatest in CT cells and least in AVR cells, contributing to their longer and shorter APD, respectively. The E-4031-sensitive rapid delayed rectifier K+ current was larger in AVR cells compared with other regions. Voltage- and time-dependent current properties were constant across regions. We conclude that myocytes from different right atrial regions of the dog show systematic variations in AP properties and ionic currents and that the spatial variation in ionic current density may explain AP differences. Regional variation in atrial ionic currents may play an important role in atrial arrhythmia generation and may present opportunities for improving antiarrhythmic drug therapy. PMID- 9734478 TI - Critical role of AT1 receptor expression after ischemia/reperfusion in isolated rat hearts: beneficial effect of antisense oligodeoxynucleotides directed at AT1 receptor mRNA. AB - To examine the relevance of angiotensin II type 1 receptor (AT1R) expression in the determination of myocardial function after ischemia/reperfusion, Sprague Dawley rats were treated intravenously with antisense oligodeoxynucleotides (AS ODNs) directed at AT1R mRNA (100 microg/rat, n=9) or scrambled antisense oligodeoxynucleotides (Scr-ODNs, 100 microg/rat, n=6). Both AS-ODNs and Scr-ODNs were given along with 300 microg/rat of liposome DOTAP/DOPE, a positive electron carrier (wt:wt= 1:1). The hearts from AS-ODN- or Scr-ODN-treated rats were excised 24 hours later, perfused in vitro, and subjected to 25 minutes of global ischemia followed by 30 minutes of reperfusion. Parallel groups of rats were given the specific AT1R antagonist losartan (10 mg/kg IV, n=6) or saline (n=7) 4 to 6 hours before excising the hearts. Ischemia/reperfusion resulted in a significant increase in myocardial AT1R expression (autoradiography and binding assay) and myocardial dysfunction, indicated by increases in coronary perfusion pressure and left ventricular end-diastolic pressure and a decrease in developed left ventricular pressure (all P<0.01 versus baseline) in the saline-treated group. AT1R protein and mRNA levels also increased in ischemic/ reperfused myocardial tissues. Administration of AS-ODNs or losartan, but not Scr-ODNs, preserved myocardial function and blocked the increased AT1R binding after ischemia/reperfusion (both P<0.01). Myocardial AT1R mRNA levels were not affected by either AS-ODNs or losartan, and the AT1R protein levels were significantly reduced by AS-ODN, but not losartan, treatment. Plasma angiotensin II levels increased after administration of losartan but not after administration of AS ODNs. These observations imply a critical role of AT1R upregulation in determining myocardial function immediately after ischemia/reperfusion. AS-ODNs to AT1R mRNA may be more beneficial than losartan, because losartan does not affect the plasma angiotensin II level. The sustained increase in AT1R mRNA, but diminished protein expression, in rat hearts treated with AS-ODNs suggests that AS-ODNs block AT1R at the translational level. PMID- 9734479 TI - Functional knockout of the transient outward current, long-QT syndrome, and cardiac remodeling in mice expressing a dominant-negative Kv4 alpha subunit. AB - A novel in vivo experimental strategy, involving cell type-specific expression of a dominant-negative K+ channel pore-forming alpha subunit, was developed and exploited to probe the molecular identity of the cardiac transient outward K+ current (I(to)). A point mutation (W to F) was introduced at position 362 in the pore region of Kv4.2 to produce a nonconducting mutant (Kv4.2W362F) subunit. Coexpression of Kv4.2W362F with Kv4.2 (or Kv4.3) attenuates the wild-type currents, and the effect is subfamily specific; ie, Kv4.2W362F does not affect heterologously expressed Kv1.4 currents. With the use of the alpha-myosin heavy chain promoter to direct cardiac-specific expression, several lines of Kv4.2W362F transgenic mice were generated. Electrophysiological recordings reveal that I(to) is selectively eliminated in ventricular myocytes isolated from transgenic mice expressing Kv4.2W362F, thereby demonstrating directly that the Kv 4 subfamily underlies I(to) in the mammalian heart. Functional knockout of I(to) leads to marked increases in action potential durations in ventricular myocytes and to prolongation of the QT interval in surface ECG recordings. In addition, a novel rapidly activating and inactivating K+ current, which is not detectable in myocytes from nontransgenic littermates, is evident in Kv4.2W362F-expressing ventricular cells. Importantly, these results demonstrate that electrical remodeling occurs in the heart when the expression of endogenous K- channels is altered. PMID- 9734480 TI - Heme oxygenase-1-derived carbon monoxide contributes to the suppression of acute hypertensive responses in vivo. AB - The enzyme heme oxygenase, which exists in inducible (HO-1) and constitutive (HO 2) isoforms, catalyzes the degradation of heme to biliverdin and CO in mammalian tissues. CO has been implicated in the control of vascular tone in a manner similar to that for NO. In the present study, we investigated the contribution of the heme oxygenase/CO pathway to the modulation of acute hypertensive responses in vivo induced by (1) alphaalphaHb, a chemically modified hemoglobin known to scavenge NO, and (2) NG-nitro-L-arginine methyl ester (L-NAME), a competitive NOS inhibitor. Experiments were carried out in conscious rats in which femoral arteries and veins were surgically catheterized 1 or 5 days before treatment with the vasoconstrictor agents. Intravenous infusion of alphaalphaHb (8% solution) or L-NAME (30 micromol/kg) [corrected] produced an acute and significant increase in mean arterial pressure (P<0.05) in rats at 5 days after catheter implantation. In contrast, no change in blood pressure was observed when alphaalphaHb or L-NAME was infused 1 day after the surgical intervention. The suppression of the hypertensive response observed at 1 day after surgery correlated with a significant (P<0.05) HO-1 expression in aorta, heart, and liver as well as increased aortic CO production and cGMP levels. At 1 day after surgery, pretreatment of animals with the heme oxygenase inhibitor zinc protoporphyrin IX (50 micromol/kg IP) markedly decreased aortic CO and cGMP levels and completely restored the vasoconstrictor effects of both alphaalphaHb and L-NAME. These results provide evidence for a crucial role of the heme oxygenase/CO pathway in the regulation of blood pressure under stress conditions in vivo. PMID- 9734481 TI - Relative importance of cytotoxic T lymphocytes and nitric oxide-dependent cytotoxicity in contractile dysfunction of rejecting murine cardiac allografts. AB - BACKGROUND: Previous in vitro studies have suggested that both cytotoxic T lymphocyte (CTL)-mediated and non-CTL-mediated myocyte lysis occur during murine cardiac heterotopic allograft rejection, but the relative importance of these injury mechanisms on myocardial function is not established. We therefore compared the in vivo effects of depletion of CTL and inhibition of nitric oxide synthase (NOS) on contractility of the rejecting heart. METHODS: Syngeneic (BALB/c into BALB/c) and allogeneic (BALB/c into C57/B16) heterotopic abdominal cardiac transplants were performed. In some of the allogeneic transplants, CD8+ lymphocytes were depleted by intraperitoneal injection of anti-CD8 monoclonal antibody. NOS inhibition was accomplished by continuous infusion of NG-monomethyl L-arginine via a subcutaneous osmotic pump. Five days after transplantation, the abdominal cavity was opened and the transplanted heart exposed. Base to apex developed force was measured during spontaneous beating at a diastolic stretch of 4 g by placing a suture through the apex of the heart and attaching it to a strain gauge. Effects of interventions on graft survival were determined by recording the days required for loss of palpable graft contractions. RESULTS: Allogeneic hearts showed a significant reduction in systolic force compared to non-rejecting syngeneic hearts. Depletion of CD8+ cells improved contractility significantly relative to non-depleted allogeneic hearts, but contractility remained significantly reduced relative to syngeneic hearts. Developed force in allogeneic hearts was also improved by NOS inhibition (P<0.01), and NG-monomethyl L-arginine infusion slightly prolonged graft survival. CONCLUSION: Both CTL mediated and NOS-dependent (possibly macrophage-mediated) mechanisms contribute to contractile dysfunction during early cardiac allograft rejection in this model. However, NOS inhibition combined with CTL depletion only slightly prolongs graft survival in this model. PMID- 9734482 TI - Estradiol 17-beta represses insulin-like growth factor I receptor expression in smooth muscle cells from rabbit cardiac recipients. AB - BACKGROUND: A crucial step in cell cycle progression is the activation of the insulin-like growth factor I (IGF-I) receptor (IGF-IR) by its ligand. Earlier, we found estradiol 17-beta treatment of cardiac allograft recipients attenuates transplant arteriosclerosis; this was associated with inhibition of vascular cell proliferation induced by IGF-I. The current study demonstrates regulation of IGF IR by estradiol 17-beta in vivo and in vitro in recipient native and allograft aorta and in aorta smooth muscle cells (SMCs). METHODS: Twenty cardiac transplant recipient rabbits were treated with estradiol 17-beta (100 microg/kg/day) or placebo for 6 weeks. IGF-IR expression in the coronary arteries of rabbits was demonstrated by immunohistochemistry. Reverse transcription-polymerase chain reaction and RNase protection assay were used to detect IGF-IR mRNA in rabbit aortas and cultured aortic SMCs in the presence or absence of estradiol 17-beta in vitro. IGF-I-induced cell proliferation was performed with the aorta explants and aorta SMCs from estradiol- or placebo-treated rabbits. RESULTS: Estradiol 17 beta treatment of rabbits significantly inhibited IGF-IR expression in the allograft coronary arteries and abrogated cell proliferation induced by IGF-I in the allograft aorta compared with placebo-treated recipients (65.4+/-5% vs. 500+/ 139%, P<0.002). Expression of IGF-IR mRNA in the allograft aorta of placebo treated recipients was significant higher than that of the native aorta (286+/ 56%, P<0.02). Estradiol treatment significantly inhibited IGF-IR mRNA expression in the aorta versus that of the placebo-treated recipients (65+/-8.5% vs. 140+/ 23%, P<0.02). Repression of IGF-IR mRNA expression in aortic SMCs by estradiol in vitro was in a concentration-dependent manner (P<0.02). CONCLUSION: Repression of IGF-IR protein and mRNA by estradiol 17-beta in vivo and in vitro suggest that one of the mechanisms of estradiol inhibition of SMC proliferation and transplant arteriosclerosis is down-regulation of IGF-IR. PMID- 9734484 TI - T cells in islet-like cell cluster xenograft rejection: a study in the pig-to mouse model. AB - BACKGROUND: The aim of the present study was to evaluate the nature of T cells involved in and, presumably, critical to fetal porcine islet-like cell cluster (ICC) xenograft rejection. METHODS: Normal mice and T cell receptor (TCR)-beta-, TCR-delta-, or TCR-betaxdelta-deficient mice were transplanted with fetal porcine ICC under the kidney capsule. Perforin- or granzyme B (GraB)-deficient mice were used to further characterize T cell-dependent pathways. For evaluation of the role of T cells in the activation process of macrophages, TCR-betaxdelta mutants were treated with recombinant mouse tumor necrosis factor (TNF)-alpha. In addition, normal mice transplanted with porcine ICC were treated with MDL 201,449A, a novel transcriptional inhibitor of TNF-alpha. RESULTS: In normal mice, the majority of the infiltrating cells were large, macrophage-like cells expressing the macrophage-specific phenotype marker F4/80. CD3+ T lymphocytes were found to be mainly accumulated in the peripheral parts of the ICC xenograft. TCR-beta mutants and TCR-betaxdelta mutants exhibited no signs of xenograft rejection, whereas TCR-delta mutants and perforin- and GraB-deficient animals rejected the ICC xenograft. Posttransplant high-dose recombinant mouse TNF-alpha treatment of TCR-betaxdelta mutants did not result in fetal porcine ICC xenograft rejection. However, a somewhat increased amount of F4/80+ and Mac-1+ cells was observed within the xenograft area. Similarly, although graft survival was not found to be prolonged, reduced numbers of CD4+ T cells were observed in mice treated with MDL 201,449A. CONCLUSIONS: In the pig-to-mouse model, fetal porcine ICC xenograft rejection is exclusively dependent on T cells bearing TCR-alphabeta chains. In addition, the absence of perforin or GraB has no influence on the rejection process, suggesting that xenospecific cytolytic T cells are of minor importance. Even if TNF-alpha is of importance to the developing process of ICC xenograft rejection, other cytokines, i.e., interferon-gamma, might efficiently substitute for the lack of TNF-alpha. PMID- 9734483 TI - Utility of adenoviral-mediated Fas ligand gene transfer to modulate islet allograft survival. AB - BACKGROUND: One of the best-defined mechanisms for the induction of apoptosis involves signaling via the cell surface molecule Fas, after binding of Fas ligand. Expression of Fas ligand is tightly regulated, being expressed primarily by T cells after activation, where it serves as a self-regulatory mechanism for immune responses. Fas ligand has also been found to be expressed constitutively at sites of immune privilege such as the testes and the anterior chamber of the eye. Recently, co-transplantation of Fas ligand-transfected myoblasts in association with islet cell allografts was shown to prolong islet allograft survival but only rarely led to indefinite graft survival. Graft rejection was associated with loss of Fas ligand on the myoblasts, suggesting that direct expression of the transgene on the islets might be more effective. METHODS: A replication-defective adenoviral construct containing murine Fas ligand (Ad/MFL) was prepared by homologous recombination. NIH 3T3 cells, rodent splenocytes, and murine islets were infected with Ad/MFL and examined in vitro for functional murine Fas ligand expression. Survival of Ad/MFL-infected islets was subsequently evaluated in vivo in both syngeneic and allogeneic islet transplantation models. RESULTS: Cell lines and islet allografts transfected with Ad/MFL expressed a functional Fas ligand, capable of inducing apoptosis (confirmed by three distinct assays for DNA fragmentation) in Fas+ targets, but not in Fas- controls. Furthermore, Ad/MFL was able to modify allogeneic immune responses in vitro, as addition of this virus, but not a control adenovirus, significantly reduced proliferation in a mixed lymphocyte reaction. Surprisingly, however, transplantation of islet allografts transfected with Ad/MFL resulted in long-term allograft survival in only 1 of 30 recipients. Moreover, adenoviral-mediated Fas ligand gene transfer was complicated by transient, dose-dependent islet dysfunction, perhaps contributing to the lack of long-term engraftment. CONCLUSION: These data suggest that adenoviral-mediated Fas ligand expression may impair normal islet function in vivo, and indicate that alternative strategies for Fas ligand transgene delivery may be required in this setting. PMID- 9734486 TI - Autotransplantation of rat parathyroid glands: a study on morphological changes. AB - BACKGROUND: Autotransplantation of parathyroid glands in man is performed to preserve parathyroid function after surgery. In a rat model, we performed autotransplantation into the renal subcapsular space to examine reinnervation and changes in cell activity in the transplanted glands. METHODS: Parathyroids grafted for 1-20 weeks were examined immunocytochemically for general and specific neuroendocrine markers to visualize nerve fibers and glandular cells and for bromodeoxyuridine to determine cell proliferation. In situ hybridization was used to localize and quantitate chromogranin A and parathyroid hormone (PTH) mRNA expression. RESULTS: Reinnervation was observed as early as 1 week after transplantation in that nerve fibers containing the general neuronal marker protein gene product 9.5 appeared along blood vessels. During the following 20 weeks, the nerve fiber density increased gradually. One week after transplantation, the immunoreaction intensity for PTH, chromogranin A, and pancreastatin was lower than in control glands. Bromodeoxyuridine-labeled cells were fewer than in control glands at 1 week and at 5-10 weeks after transplantation. The density of PTH mRNA labeling was lower than in control glands during the whole time period studied and reached a minimum after 10 weeks. The density of chromogranin A mRNA labeling was unaffected at 1 and 3 weeks after transplantation and then decreased to a minimum at 10 weeks after transplantation; at 20 weeks, the chromogranin A mRNA labeling had again reached the level in control glands. CONCLUSION: The changes in PTH and chromogranin A immunoreaction intensity and mRNA density indicate reduced hormone production for several weeks after transplantation. Our results using transmitter-specific markers indicate a rapid ingrowth of mostly sympathetic nerve fibers, preferentially around blood vessels. Later on, parasympathetic and sensory nerve fibers reached the grafts. The parathyroid innervation may be of importance for parathyroid hormone regulation, and the finding of an early reinnervation could be of clinical importance. PMID- 9734485 TI - Fatty acid omega and (omega-1)-oxidation within intrasplenically transplanted fetal hepatocytes. AB - BACKGROUND: The expression and enzymatic activity of the cytochrome P450 LAomega within intrasplenically transplanted hepatocytes was investigated. METHODS: Fetal hepatocytes were harvested from spontaneously hypertensive rats and transplanted into recipient adult spontaneously hypertensive rat spleens. RESULTS: Microscopic examination revealed masses of hepatocytes in the red pulp. Immunochemical studies detected cytochrome P450 LAomega in transplanted hepatocytes by 6 and 10 weeks after transplantation. Cytochrome P450 LAomega mRNA accumulates at 6 weeks after transplantation. Cytochrome P450-arachidonic acid omega/omega-1 hydroxylase activity (formation of 20/19-hydroxyeicosatetraenoic acid) was detected at 10 weeks after transplantation. CONCLUSION: These results demonstrated that fetal hepatocytes grow in the spleen and function similarly to adult hepatocytes. PMID- 9734487 TI - Mechanism of tolerance to class I-mismatched renal allografts in miniature swine: regulation of interleukin-2 receptor alpha-chain expression on CD8 peripheral blood lymphocytes of tolerant animals. AB - BACKGROUND: Donor-specific tolerance to renal allografts in miniature swine is uniformly induced across a two-haplotype class I plus minor histocompatibility antigen disparity by a 12-day course of cyclosporine. Recent studies have demonstrated that the thymus is essential for rapid and stable tolerance induction, because either prior thymectomy or a series of thymic biopsies induce a spontaneously reversible rejection crisis after the 12-day course of cyclosporine. The present study examined the peripheral cellular mechanisms of tolerance by analyzing cytotoxic effector pathways in peripheral blood lymphocytes (PBL) of tolerant animals. METHODS: The phenotype and cytotoxic T lymphocyte response of alloantigen-activated PBL cultures using cells from a series of tolerant animals with stable renal function (no thymic manipulation), or during a rejection crisis (induced by thymic biopsies), were studied. The in vitro findings were correlated with the in vivo clinical course of experimental animals. RESULTS: The data demonstrated that in vivo and in vitro tolerance was associated with a specific deficiency of interleukin-2 receptor (IL-2R) alpha chain up-regulation on CD8 single-positive (SP) T cells expressing high levels of CD8 (CD8high) when PBL from tolerant animals are stimulated with donor class I alloantigen. Stimulation by third party class I alloantigen, or by donor antigen during a rejection crisis, produced efficient cytotoxic T lymphocyte responses and expression of IL-2Ralpha on CD8high SP cells. CONCLUSION: Antigen-specific regulation of the IL-2Ralpha expression on CD8high SP PBL is a principal event associated with and potentially involved in the mechanism of tolerance in this preclinical large animal model. PMID- 9734488 TI - Delayed graft function does not reduce the survival of renal transplant allografts. AB - BACKGROUND: The aim of the present study was to investigate the effect of delayed graft function (DGF) in graft outcome when adjusted by the presence of acute rejection in the first month after transplantation. METHODS: A total of 437 cadaveric renal transplant patients on cyclosporine and steroids were included in the study. Variables related to donor, recipient, and graft were prospectively collected. RESULTS: The incidence of DGF was 44.4%. When patients dying with a functioning graft were censored, graft survival rates at 1 and 6 years were similar in patients with immediate function to those with DGF, when rejection was not present (96% and 81% vs. 95% and 83%, respectively). Rejection negatively influenced graft survival rates at 1 and 6 years, both in patients with immediate graft function (80% and 73%, P<0.05 vs. no DGF/no rejection) and more deeply in those with associated DGF (77% and 62%, P<0.001 vs. no DGF/no rejection). Rejection was more frequently diagnosed in patients with DGF than in those with immediate graft function (50% vs. 39.9%, P<0.05). Length of hospitalization was longer and the number of needle core biopsies was higher in patients with DGF or rejection. The presence of both complications had an additive effect. CONCLUSIONS: This study showed that DGF did not adversely affect kidney graft survival in patients without rejection. However, it increased the length of hospitalization and the number of graft biopsies, thus increasing the cost of transplantation. Moreover, rejection was more frequent in patients with DGF, and it had a negative impact on graft outcome. Because the association of DGF and rejection gave the poorest outcome, an effort should be made to prevent both complications. PMID- 9734489 TI - Clinical implications of the diagnosis of renal allograft infarction by percutaneous biopsy. AB - BACKGROUND: Herein we investigated the relationships between acute rejection (AR), infection, and renal allograft infarcts, particularly those infarcts that occur beyond the immediate posttransplant period and that affect functioning grafts. METHODS: Infarcts (n=59) were classified as: (1) early (EI; <2 months after transplant; n=32); or (2) late (LI; >2 months; n=27). Controls included patients with severe AR but without infarction (n=84). RESULTS: There were not significant differences in donor or recipient characteristics between infarcts and controls. At diagnosis, patients with infarcts were more likely to be infected (30%) than controls (14%, P=0.01); 15% of infarcts and 1% of controls had disseminated cytomegalovirus (P=0.04). Infarct and AR coexisted in the biopsy specimens of 66% of patients with EI and 62% of patients with LI, but the AR severity ranged from borderline to severe. Furthermore, 30% of patients with EI/LI had a history of severe AR. Graft survival was 47% in patients with EI, 22% in patients with LI (NS), and 71% in controls (P<0.0001, chi-square and Cox regression). Correlates of better graft survival in infarcts included: older recipient (P=0.03); smaller area of infarction in the biopsy specimen (P=0.04); and use of anti-AR therapy (P=0.03). Therapy was effective in patients with EI (treated, 71% survival; untreated, 29%, P=0.02) but not in patients with LI (25% vs. 23%). CONCLUSIONS: Allograft infarcts are associated with AR in 64% of patients, but the AR may be mild. Infarcts are associated with infections. Graft survival is worse in patients with infarcts than in patients with severe AR, consequently these two pathologic diagnoses should not be considered as a single entity. PMID- 9734490 TI - Retrospective study on the impact of hepatitis C virus infection on kidney transplant patients over 20 years. AB - BACKGROUND: The majority of chronic hepatitis is ascribable to hepatitis C virus (HCV) infection, whereas the clinical impact has not been understood in kidney transplant recipients. Our current study was carried out to assess the impact of HCV infection on kidney recipients over the long-term, and to investigate the effect and risk of interferon-alpha (IFN-alpha) therapy for chronic active hepatitis C. METHODS: Hepatitis B surface antigen (HBsAg) and antibody to HCV (HCVAb) were examined prospectively and retrospectively in 280 patients, who underwent kidney transplants in the period from 1973 to 1996. The patient survival rate, the graft survival rate, the incidence of liver dysfunction and the cause of mortality among the HCV infected and noninfected groups were analyzed. IFN-alpha therapy was performed on 10 patients with chronic active hepatitis C. RESULTS: Prevalence of the hepatitis virus was quite high at 34.3% (96/280): the frequency of the HBsAg carrier was 3.2% (9/280), that of the HCVAb carrier was 28.6% (80/280) and that of the both carriers was 2.5% (7/280). The other 184 cases (65.7%) were negative for both HBsAg and HCVAb. Liver dysfunction developed at the significantly higher incidence of 55% in HCVAb carriers compared to the 9.2% of the noninfected group (P<0.01). HCVAb carriers had a poor survival rate in the second decade compared to the noninfected group: 83.7% vs. 88.91% for 10-year survival (P=0.44) and 63.9% vs. 87.9% for 20-year survival (P<0.05). The poor survival rate was a result of the mortality from liver disorder. Five patients died of such disease in the infected groups whereas no noninfected patient died in the same period (p<0.01). As the result of IFN-alpha therapy, biochemical activity normalized or improved in eight cases, whereas the HCV-RNA titer was reduced only in three patients. Only one patient maintained normal biochemical markers and undetectable levels of HCV-RNA for 2 years after treatment. The therapy was discontinued for five patients with the adverse effects of acute rejection, deterioration of diabetes, and depression. CONCLUSIONS: HCV infection has a significant impact on kidney transplant recipients over the long term and in particular affects them in the second decade. Our pilot study revealed only partial efficacy of IFN-alpha therapy for HCV-infected recipients, but with the high risk of acute rejection. PMID- 9734491 TI - Prostaglandin E1 analogs do not improve renal function among either transplant or nontransplant patients: no further trials required. AB - BACKGROUND: To assess whether prostaglandin E1 analogs have a role in either reducing renal allograft rejection or improving renal function among both transplant and nontransplant patients. METHODS: Studies were identified through Ovid MEDLINE between 1981 and December 1997 using multiple MeSH headings and text words related to renal or liver transplantation, as well as renal insufficiency. These items were crossed with MeSH headings and text words related to prostaglandins and prostaglandin E1. All abstracts were read, in addition to review articles, and their bibliographies were searched for further references. Articles were limited to those published in the English language. Studies were selected based on the following criteria: (1) a randomized control clinical trial; (2) administration of any form of prostaglandin; (3) publication of primary data; and (4) reporting on either renal transplant rejection or renal dysfunction after transplant or, for both transplant and nontransplant studies, objectively comparing a change in renal function from before to after prostaglandin E1 therapy. From the 217 articles that were retrieved, 19 met all inclusion criteria. Data were extracted on study design, nature of the study subjects, and the principal therapeutic intervention. Among the transplant studies, the rate of acute renal graft rejection or renal dysfunction was calculated for each study and then pooled using a random effects model. In addition, the mean change in renal glomerular filtration rate was compared between prostaglandin E1 and controls among both transplant and nontransplant studies and then pooled using an inverse variance-weighted method. Using the Breslow-Day method, statistical heterogeneity was defined at a two-sided P value less than 0.10. RESULTS: Within the 10 transplant trials, all patients were on cyclosporine; oral or intravenous prostaglandin E1 was generally started within 24 hr of transplantation. Renal transplant rejection or renal dysfunction was not significantly reduced with prostaglandin E1 (odds ratio 0.91, 95% confidence interval [CI] 0.64 to 1.28, two-sided P value=0.58; weighted control event 66.9%, 95% CI 50.5 to 80.0). The glomerular filtration rate was estimated within nine transplant studies, with a minimal positive gain in renal function with prostaglandin E1 (mean difference 2.3 ml/min, 95% CI 1.6 to 3.1). Nine other randomized, double-blinded trials evaluated the effect of prostaglandin E1 on renal function among a variety of nontransplant patients. These patients were generally selected based on their susceptibility to renal injury, with a concomitant exposure to nonsteroidal anti-inflammatory drugs. No significant change in the glomerular filtration rate was observed between prostaglandin E1 and placebo arms (mean difference 0.5 ml/min in favor of placebo, 95% CI -2.8 to 1.8). However, these studies were very heterogeneous. CONCLUSIONS: Among both transplant and nontransplant populations, prostaglandin E1 does not seem to preserve or improve renal function. Further research is unlikely to demonstrate superiority of prostaglandin E1 in the context of cyclosporine or nonsteroidal anti-inflammatory drug use. PMID- 9734492 TI - Auxiliary partial orthotopic liver transplantation from living donors: significance of portal blood flow. AB - BACKGROUND: Auxiliary liver transplantation has several advantages over standard orthotopic liver transplantation. However, functional competition has been reported even in auxiliary partial orthotopic liver transplantation (APOLT). We evaluated herein the interaction in APOLT between the native liver and the graft in terms of portal blood flow and regeneration. The need for diversion of the portal blood flow to the graft was also assessed. METHODS: A total of 15 patients received APOLT from living donors. Portal blood flow to the native liver was preserved in 6 patients, and the portal vein to the native liver was preemptively transected at the time of transplantation in 9 patients. RESULTS: Of the patients with preservation of the portal blood flow to the native liver, two showed inadequate graft portal blood flow just after operation, and in the other three patients the graft portal blood flow decreased or the graft atrophied after deterioration of the graft function. In the patients with preemptive transection of the portal vein to the native liver, optimal graft portal blood flow was obtained, and the native liver, supplied only by arterial inflow, supported a small-for-size graft until the graft regenerated. The damage to the native liver was minimal. CONCLUSIONS: Functional competition may occur in APOLT with preservation of the portal blood flow to the native liver, whereas preemptive transection of the native liver portal vein is a safe procedure and effectively prevents the portal steal phenomenon. PMID- 9734493 TI - Reduced-size orthotopic composite liver-intestinal allograft. AB - A composite graft consisting of a reduced left lateral hepatic segment in continuity with the small intestine was procured from an adult cadaveric donor using a modified in situ split technique. The primary recipient was a 3-year-old boy with hepatointestinal failure. The right side of the liver was transplanted into a 63-year-old man with a central hepatoma and hepatitis C cirrhosis. This was accomplished with center-to-center sharing of the liver portion of the allograft. The in situ split technique was feasible, with good initial allograft function. However, both grafts failed subsequently because of peri-operative recipient-related complications. The adult patient died of an infected pseudoaneurysm of the arterial graft, and the pediatric patient required repeat transplantation as a result of the late diagnosis of a native pancreatic fistula with cholestatic damage to the reduced liver allograft. The child is currently alive 8 months after repeat transplantation. PMID- 9734494 TI - A long-term comparison of tacrolimus (FK506) versus cyclosporine in liver transplantation: a report of the United States FK506 Study Group. AB - BACKGROUND: The long-term (5 year) efficacy and safety of tacrolimus (FK506) and cyclosporine were compared in primary liver transplant recipients who participated in a 1-year randomized, multicenter trial and a 4-year follow-up extension study. METHODS: A total of 529 patients (263 tacrolimus group, 266 cyclosporine group) were randomized to study drug. Patients were evaluated at 3 month intervals. Patient and graft survival rates, incidence of adverse events, and changes in laboratory and clinical profiles were determined. RESULTS: Cumulative 5-year patient and graft survival rates were comparable for the tacrolimus (79.0%, 71.8%) and cyclosporine (73.1%, 66.4%) groups. However, patient half-life survival was longer for tacrolimus-treated patients (25.1+/-5.1 years versus 15.2+/-2.5 years; P=0.049). Improved patient survival with tacrolimus was also observed for hepatitis C-positive patients (78.9% tacrolimus group versus 60.5% cyclosporine group; P=0.041). Both treatments were associated with a low incidence of late acute rejection, late steroid-resistant rejection, and death or graft loss related to rejection. Both treatments demonstrated an acceptable safety profile with maintenance of adequate renal and liver function and a low incidence of malignancy/lymphoproliferative disease and serious infections. CONCLUSIONS: Tacrolimus is a safe and effective long-term maintenance immunosuppressive agent in primary liver transplantation. PMID- 9734495 TI - Age and liver transplantation: a report of the Liver Transplantation Database. AB - BACKGROUND: The average age of liver transplant recipients has increased steadily during the last decade. The effects of recipient age on outcome of liver transplantation were evaluated in a large prospective database. METHODS: A total of 735 adult recipients of single-organ liver transplants for nonfulminant liver disease enrolled in a large prospective database between 1990 and 1994 were analyzed for associations of patient age with outcomes. Patients were categorized into two groups: younger being <60 and older being > or = 60 years of age. RESULTS: Older liver transplant recipients were more likely to be female, white, and have the diagnoses of primary biliary cirrhosis or cryptogenic cirrhosis than younger recipients, who were more likely to have the diagnosis of alcoholic liver disease. Disease severity was similar between the two groups. After transplantation, the durations of stay in the intensive care unit and hospital were longer for older than for younger transplant recipients, but episodes of acute rejection were less frequent. The quality of life at 1 year was similar among older and younger recipients. Patient survival was lower for older than for younger recipients (81% vs. 90% at 1 year; P=0.004), whereas graft survival was not different (80% vs. 85% at 1 year; P=0.163). The excess mortality among older recipients was largely due to nonhepatic causes, including infectious, cardiac, and neurological diseases occurring within 6 months after transplantation. CONCLUSIONS: Although patient survival was significantly lower among liver transplant recipients above the age of 60 years, the excess mortality was due to nonhepatic, largely age-related problems. The overall success of liver transplantation and improvement in quality of life for older recipients is excellent. PMID- 9734496 TI - A randomized active-controlled trial of mycophenolate mofetil in heart transplant recipients. Mycophenolate Mofetil Investigators. AB - BACKGROUND: After heart transplantation, 1-year and 5-year survival rates are 79% and 63%, respectively, with rejection, infection, and allograft coronary artery disease accounting for the majority of deaths. Mycophenolate mofetil (MMF), an inhibitor of the de novo pathway for purine biosynthesis, decreases rejection in animals and in human renal transplantation. METHODS: In a double-blind, active controlled trial, 28 centers randomized 650 patients undergoing their first heart transplant to receive MMF (3000 mg/day) or azathioprine (1.5-3 mg/kg/day), in addition to cyclosporine and corticosteroids. Rejection and survival data were obtained for 6 and 12 months, respectively. Because 11% of the patients withdrew before receiving study drug, data were analyzed on all randomized patients (enrolled patients) and on patients who received study medications (treated patients). RESULTS: Survival and rejection were similar in enrolled patients (MMF, n=327; azathioprine, n=323). In treated patients (MMF, n=289; azathioprine, n=289), the MMF group compared with the azathioprine group was associated with significant reduction in mortality at 1 year (18 [6.2%] versus 33 deaths [11.4%]; P=0.031) and a significant reduction in the requirement for rejection treatment (65.7% versus 73.7%; P=0.026). There was a trend for fewer MMF patients to have > or = grade 3A rejection (45.0% versus 52.9%; P=0.055) or require the murine monoclonal anti-CD3 antibody or antithymocyte globulin (15.2% versus 21.1%; P=0.061). Opportunistic infections, mostly herpes simplex, were more common in the MMF group (53.3% versus 43.6%; P=0.025). CONCLUSIONS: Substitution of MMF for azathioprine may reduce mortality and rejection in the first year after cardiac transplantation. PMID- 9734497 TI - Donor-specific hyporeactivity after liver transplantation: prominent decreases in donor-specific cytotoxic T lymphocyte precursor frequencies independent of changes in helper T lymphocyte precursor frequencies or suppressor cell activity. AB - BACKGROUND: The development of immunological donor-specific hyporeactivity may account for the low incidence of chronic rejection after clinical liver transplantation. We investigated whether hyporeactivity commonly develops after liver transplantation by analyzing precursor frequencies of donor-reactive cytotoxic (CTLp) and helper (HTLp) T lymphocytes and mixed lymphocyte culture (MLC) reactivity in liver allograft recipients. We further studied whether CTLp hyporeactivity correlated with changes in donor-specific HTLp frequencies or suppressor cell activity. METHODS: CTLp and HTLp frequencies and MLC reactivity against donor and third-party spleen cells were determined in pre- and posttransplantation peripheral blood samples from 18 recipients with good graft function 2 years after transplantation. By mixing posttransplantation samples (with "putative" suppressor cell activity) with pretransplantation samples (in which normal CTL activity with no suppressor cell activity is expected), the presence of suppressor cell activity in peripheral blood was analyzed. RESULTS: Two years after transplantation, all but one (94%) of the recipients had developed CTLp hyporeactivity as evidenced by reduced donor-specific CTLp frequencies. The development of hyporeactivity was not specific for any particular underlying disease. The occurrence of HTL hyporeactivity, however, was less frequent: 38% and 20% of recipients were HTLp and MLC hyporeactive, respectively. Decreases in CTLp frequencies did not correlate with decreased donor-specific HTL function or suppressor cell activity in peripheral blood samples. CONCLUSIONS: Donor-specific CTLp hyporeactivity can develop in the majority of liver allograft recipients, irrespective of underlying disease. Donor specific HTL hyporeactivity, however, occurs infrequently. A reduction in donor specific CTLp frequencies was found to be independent of changes in donor specific HTLp or suppressor cell activity, suggesting that other mechanisms (e.g., clonal deletion) are operative in the reduction of donor-specific CTLp after liver transplantation. PMID- 9734498 TI - A strong impact of matching for a limited number of HLA-DR antigens on graft survival and rejection episodes: a single-center study of first cadaveric kidneys to nonsensitized recipients. AB - BACKGROUND: A single-center study of 655 nonsensitized recipients of primary cadaveric kidney grafts is presented. RESULTS: Graft survival in serologically HLA-DR 1-10 antigen-matched grafts to nonsensitized recipients at 1 year was 90%, compared with 82% (P=0.004) and 73% (P=0.001) in one and two DR antigen mismatched grafts. The corresponding figures at 5 years were 76%, 62%, and 56%, respectively. Matching for the DR antigens 11-14, or for some DR alleles only detectable by genomic typing, further improved graft survival, but the differences did not reach statistical significance. Matching also for the serologically defined HLA-A and -B antigens did not significantly further improve overall graft survival, but some effects for grafts surviving at least 1 year were observed. Among recipients of grafts mismatched for zero, one, or two HLA-DR antigens, acute rejection episodes were experienced in 48%, 64% (P<0.001), and 82% (P<0.001), respectively, within the first 3 months. HLA-A and -B mismatches showed no significant correlation to acute rejection episodes. CONCLUSION: Matching for the DR antigens 1-10 significantly secures and prolongs the survival of first cadaveric renal grafts. Our results also show that DR 1-10 antigen matched combinations can often be obtained even in rather small recipient pools, when actively sought for. PMID- 9734499 TI - Liver and kidney transplantation for polycystic disease. AB - BACKGROUND: With the poor results of resective and fenestration procedures for polycystic liver disease (PCLD), we present the first series of patients receiving orthotopic liver transplantation for this condition. METHODS: Five of our six patients with PCLD had polycystic kidney disease also. Three of these five received combined organ transplants, while the other two required subsequent kidney transplants. RESULTS: Forty-eight and 52 months after orthotopic liver transplantation, all surviving patients had relief of their pain, distention, and anorexia. Two patients had succumbed to infectious complications and died at 15 and 24 months after transplant. CONCLUSIONS: We conclude that patients with PCLD can be transplanted safely for the relief of their distention and anorexia, with good results. Those patients with both PCLD and polycystic kidney disease who are not dialysis dependent can be managed for several years with isolated liver transplantation and then receive kidney transplantation if needed. Those who are dialysis dependent should receive combined liver-kidney transplantation. Unfortunately, patients with polycystic disease seem to be very susceptible to infectious complications after organ transplantation. PMID- 9734500 TI - Efficacy and safety of low molecular weight heparin in renal transplantation. AB - BACKGROUND: Deep venous thrombosis (DVT) is a common problem with potentially devastating results in patients undergoing major surgical procedures. Certain renal transplant recipients are particularly at risk for allograft loss as a consequence of renal vein and artery thrombosis. Over the past few years, low molecular weight heparin has been well established as an accepted modality of treatment and prophylaxis of DVT. The efficacy and safety of low molecular weight heparin in the prophylaxis of DVT following renal transplantation in adults has not previously been reported. METHODS: Dalteparin was administered to 120 adult renal transplant recipients postoperatively at the Oregon Health Sciences University. RESULTS: No patient developed allograft arterial or venous thrombosis. One patient developed subclavian vein thrombosis. No bleeding complications were encountered, and side effects were very minimal. CONCLUSION: Prophylaxis with dalteparin is an effective and safe modality for the prevention of thrombosis in adult patients undergoing renal transplantation. PMID- 9734501 TI - QT prolongation and near fatal cardiac arrhythmia after intravenous tacrolimus administration: a case report. AB - BACKGROUND: The use of immunosuppressant agents is mandatory in the long-term management of transplant recipients. Herein, we report a case of near fatal cardiac arrhythmia related to the use of intravenous tacrolimus in a 35-year-old woman undergoing renal transplantation. METHODS: The patient had no previous history of cardiac disease, but an initial electrocardiogram demonstrated slightly prolonged QT and QTc intervals and normal sinus rhythm. Postsurgical immunosuppression included intravenous tacrolimus and methylprednisolone. During intravenous tacrolimus infusion, marked QT prolongation occurred. The patient suffered recurrent runs of torsade de pointes, refractory to aggressive medical management and requiring numerous defibrillations. Rapid atrial pacing eventually controlled the arrhythmia. RESULTS: We note not only a temporal association, but also a direct linear relationship, between this arrhythmia and blood tacrolimus levels. CONCLUSION: We believe this case presents a little recognized hazard associated with the use of intravenous tacrolimus and points to the need for careful predrug screening for QT prolongation. Tacrolimus has been shown to effect intracellular calcium and to prolong the action potential duration experimentally. This suggests that an increase in the intracellular calcium may underlie torsades de pointes associated with intravenous tacrolimus. PMID- 9734502 TI - Night blindness secondary to vitamin A deficiency in a patient with bile duct strictures after liver transplantation. AB - Vitamin A deficiency and resulting night blindness have previously been reported in patients with chronic liver disease before undergoing liver transplantation. Because early identification of patients with vitamin A deficiency can lead to the relief of symptoms and the prevention of irreversible retinal degeneration, vitamin A deficiency should always be considered in the differential diagnosis of visual disturbances in patients with liver disease. We describe a case of night blindness due to vitamin A deficiency resulting from bile duct strictures in a post-orthotopic liver transplant patient and its successful resolution with vitamin A supplementation. PMID- 9734503 TI - Canine T cells transduced with a herpes simplex virus thymidine kinase gene: a model to study effects on engraftment and control of graft-versus-host disease. AB - BACKGROUND: Alloreactive donor T cells in marrow grafts mediate graft-versus-host disease (GVHD), but T-cell depletion has resulted in increased graft failure. Add back of gene-modified alloreactive donor T cells could prevent graft rejection. After engraftment, in vivo depletion of those modified T cells with ganciclovir may control GVHD. METHODS: Canine recipient-specific donor cytotoxic T lymphocytes (CTL) were retrovirally transduced with the herpes simplex virus thymidine kinase gene. RESULTS: Gibbon ape leukemia virus-pseudotyped vector yielded primary CTL transduction efficiency of 22.9+/-9.9%. After selection and expansion, 96.7+/-0.8% of CTL expressed retrovirally transferred genes. Recipient specific cytotoxic activity was maintained with 84.3% specific lysis. After ganciclovir treatment, herpes simplex virus thymidine kinase-transduced CTL proliferation was reduced 98.7+/-0.2% compared with controls. CONCLUSIONS: We have demonstrated efficient ex vivo transduction, expansion, maintenance of alloreactivity, and ganciclovir-mediated ablation of canine CTL, which will permit in vivo studies in the dog, a well-established model for GVHD and engraftment. PMID- 9734504 TI - Matters of life and death: the challenge of CPR decision making: how can we improve patient involvement in this complex process? PMID- 9734505 TI - Twenty-four hour access to health information and advice: an essential component of the healthcare system. PMID- 9734506 TI - General practice stress: winds of change buffet general practitioners. PMID- 9734508 TI - The stress of metropolitan general practice. AB - OBJECTIVE: To identify the work-related stressors of Australian metropolitan general practitioners (GPs). DESIGN AND SETTING: A descriptive postal survey of metropolitan GPs from all States and Territories selected at random from the Health Insurance Commission database. PARTICIPANTS: 296 of 464 GPs (64%) surveyed in June 1996; 67% were male; 87% worked full-time (more than 6 sessions per week). MAIN OUTCOME MEASURES: Frequency and severity of work stressors in general practice; overall feelings of stress at work in the past 12 months; effects of the stressors on work satisfaction; contribution of work stress to overall life stress; responses to the 12-item General Health Questionnaire (GHQ) as potential correlates of occupational stress. RESULTS: "Time pressure to see patients" was the most frequently reported stressor. Threat of litigation was perceived as the most severe stressor. Of the top 10 severe stressors, seven were also in the top 10 for stressor frequency. Work was the major stressor in GPs' lives. The GHQ scores did not correlate significantly with major stress outcome measures, but 12.8% of GPs had scores indicative of severe psychiatric disturbance. Fifty per cent of respondents had considered leaving their current workplace and 53% had considered abandoning general practice because of occupational stress. GPs working 6 or more sessions per week were more likely to be moderately or severely stressed than those working part-time (P< 0.02, Fisher's exact test). Those who had considered leaving their current workplace or careers were also more likely to be moderately or severely stressed (P< 0.0001, Fisher's exact test). CONCLUSIONS: The most frequent and relatively severe stressful events in general practice involved time pressures. There are implications for government, which, through remuneration policies, might influence GPs to work at a rate beyond their capacity to cope. Strategies are required to manage or prevent stress in metropolitan GPs. PMID- 9734507 TI - Decision making in CPR: attitudes of hospital patients and healthcare professionals. AB - OBJECTIVE: To examine the opinions of patients and healthcare professionals regarding the process of making decisions about cardiopulmonary resuscitation (CPR). DESIGN AND PARTICIPANTS: A cross-sectional survey of 511 healthcare professionals (doctors, nurses and allied health professionals) (64% response rate) and 152 patients (58% response rate) at the John Hunter Hospital, Newcastle, New South Wales, in June 1994. MAIN OUTCOME MEASURES: Opinions on who should be involved in CPR decision making; what issues are important when making the decision; and how these decisions should be communicated. RESULTS: 80% (95% confidence interval [CI], 72%-86%) of patients and 99% (95% CI, 98%-100%) of healthcare professionals (P<0.001) thought patients' views should be taken into account when making CPR decisions. More patients (29%; 95% CI, 22%-38%) than healthcare professionals (14%; 95% CI, 11%-17%) indicated that doctors should be the main decision makers. Two-thirds of respondents regarded the patient's wishes, diagnosis and quality of life as important factors. Most respondents (82%) felt comfortable discussing CPR, but only 29% (95% CI, 22%-37%) of patients and 57% (95% CI, 52%-61%) of healthcare professionals had actually discussed CPR with others (P<0.001). More than half of all respondents preferred to express their wishes about CPR in writing (47% [95% CI, 39%-55%] of patients, 69% [95% CI, 64%-73%] of healthcare professionals; P<0.01); the others preferred to tell a family member or close friend. Most patients (60%; 95% CI, 52%-68%) and healthcare professionals (85%; 95% CI, 81%-88%) wanted their views in their medical records (P< 0.001). CONCLUSION: Most patients want to be involved in CPR decision making and many want some form of advance directive. Although there are some differences in opinions between patients and healthcare professionals, both perceive decision making at the end of life as a shared process, primarily involving the patient and doctor. PMID- 9734509 TI - Hip fracture in elderly men: the importance of subclinical vitamin D deficiency and hypogonadism. AB - OBJECTIVE: To determine the major risk factors for hip fracture in elderly men. DESIGN: Prospective recruitment, followed by analysis of clinical and biochemical variables. PATIENTS AND SETTING: Men aged 60 years and older who presented to St George Hospital (a 650-bed tertiary-care centre) in 1995, comprising all 41 men with hip fractures, as well as 41 hospital inpatient and 41 outpatient control subjects without hip fractures. MAIN OUTCOME MEASURES: Osteoporotic risk factors (including age, body weight, comorbid illnesses, alcohol intake, cigarettes smoked, and corticosteroid use) and serum concentrations of creatinine, urea, calcium, albumin, alkaline phosphatase, parathyroid hormone, 25-hydroxyvitamin D and free testosterone. RESULTS: There were no significant differences between the hip fracture and two control groups on any of the osteoporotic risk factors. Men with hip fracture had significantly lower mean serum 25-hydroxyvitamin D concentration (45.6 nmol/L; 95% confidence interval [CI], 36.9-52.3 nmol/L) than both inpatient (61.1 nmol/L; 95% CI, 50.0-72.2 nmol/L) and outpatient (65.9 nmol/L; 95% CI, 59.0-72.8 nmol/L) controls (P=0.007). Subclinical vitamin D deficiency (defined as <50 nmol/L serum 25-hydroxyvitamin D) was 63% in the fracture group, compared with 25% in the control groups combined (odds ratio, 3.9; 95% CI, 1.74-8.78; P=0.0007). Inpatients with and without hip fractures had significantly lower mean serum albumin, calcium and free testosterone concentrations than outpatients (P< 0.05). In a multiple regression analysis, subclinical vitamin D deficiency was the strongest predictor of hip fracture (beta [regression coefficient], 0.34+/-0.19; P=0.013). CONCLUSIONS: Subclinical vitamin D deficiency in Australian men may contribute significantly to the development of hip fracture through the effects of secondary hyperparathyroidism, resulting in increased bone loss. PMID- 9734511 TI - Towards general practice-led integrated healthcare in New Zealand. AB - New Zealand's recent painful experience of health system reforms has shown that professional incentives are more powerful than market incentives, and that medical leadership, with accountability for both cost and quality, may be the key to success. PMID- 9734510 TI - Emergency department telephone advice. AB - OBJECTIVE: To evaluate telephone advice given in an emergency department. DESIGN: Prospective, observational study. SETTING: A community-based emergency department in a semi-rural/outer metropolitan setting, between August and November 1995. PARTICIPANTS: All people telephoning the emergency department for medical advice. METHODS: Details of all calls, callers and patients were recorded. Within 72 hours, a follow-up call was initiated seeking replies to a series of standardised questions. MAIN OUTCOME MEASURES: Number, timing and duration of calls; appropriateness of the advice given; compliance with the advice; and callers' satisfaction with the service. RESULTS: Over the four-month period, 1682 calls were received, 58% between 4pm and midnight. There were 33 telephone calls per 100 emergency department attendances. The mean call duration was 3.9 minutes (range, 0.25-25 minutes); 49% of patients were less than 14 years old, and 72% of callers phoned because of spontaneous illness. The advice given was considered inappropriate in only 1.4% of calls. Follow-up calls were made to 1132 people (67%), revealing a non-compliance rate of only 6.9% and a high level of caller satisfaction, with 99% of callers affirming a need for such a service. CONCLUSIONS: The provision of telephone advice by emergency department staff is rated highly by the community and compliance with the advice is strong. Paediatric problems, arising as a result of spontaneous illness, predominate and there is a large bias towards after-hours use of the service. Experienced staff provide better advice. PMID- 9734512 TI - Leptospirosis associated with severe pulmonary haemorrhage in Far North Queensland. AB - Pulmonary haemorrhage as a manifestation of leptospirosis is rarely diagnosed in developed countries. Five patients with proven leptospirosis associated with severe pulmonary haemorrhage presented to one hospital in Far North Queensland between January 1994 and June 1997. Four required admission to the intensive care unit and one patient died. Pulmonary haemorrhage is an uncommon but severe complication of leptospirosis and may be a source of diagnostic confusion in tropical areas of Australia. PMID- 9734513 TI - The burden of disease. PMID- 9734514 TI - A comparison of the diseases caused by Ross River virus and Barmah Forest virus. AB - Barmah Forest virus (BFV) and Ross River virus (RRV) are mosquito-borne viruses with similar vectors and environmental requirements. They cause diseases characterised by arthralgia, arthritis and myalgia, often accompanied by fever and rash. Arthritis is more common and more prominent in RRV disease and rash is more common and florid with BFV infection, although the diseases cannot be reliably distinguished by their clinical symptoms. Diagnosis is based on serological tests and a definite diagnosis of recent infection requires the demonstration of rising titres of IgG. Arthralgia, myalgia and lethargy may continue for at least six months in up to half of patients with RRV, but in only about 10% of patients with BFV. Both diseases are managed symptomatically. PMID- 9734515 TI - Gallstones. AB - Lifetime risk of gallstones in Australia is 14%-20%. The most common symptom of gallstones in either the gallbladder or bile duct is epigastric to right upper quadrant pain. Cholecystectomy is indicated for symptomatic gallbladder stones and is usually performed laparoscopically; surgery is rarely indicated for asymptomatic stones. Endoscopic retrograde cholangiopancreatography is indicated for complications of bile duct stones, such as jaundice, cholangitis or severe pancreatitis, and for postcholecystectomy symptoms of stones; bile duct stones may be removed in the same procedure via endoscopic sphincterotomy. Bile duct stones are mostly diagnosed by operative cholangiography at cholecystectomy. Open surgery is used when minimal-access techniques are dangerous, unsuitable or impossible. PMID- 9734516 TI - The health consequences of unemployment: the evidence. PMID- 9734517 TI - Preventable hospitalisation from underuse of drugs. PMID- 9734518 TI - Coronary risk factors 6-12 months after coronary artery bypass surgery. PMID- 9734519 TI - Bat lyssavirus prophylaxis in an immunocompromised patient. PMID- 9734520 TI - Death is a journey to be undertaken. PMID- 9734521 TI - Death is a journey to be undertaken. PMID- 9734522 TI - Death is a journey to be undertaken. PMID- 9734523 TI - Choking after inhaling a foreign body through a Ventolin puffer. PMID- 9734524 TI - Choking after inhaling a foreign body through a Ventolin puffer. PMID- 9734525 TI - Zoonoses. PMID- 9734526 TI - Survival and disability at 7-8 years of age in New Zealand infants less than 28 weeks gestation. AB - AIMS: To determine the survival and disability rates at 7-8 years in infants of less than 28 weeks gestation born in New Zealand in 1986 and admitted to a neonatal unit. METHODS: In 1986, all infants with birthweight less than 1500 g and admitted to neonatal units were enrolled in a prospective audit of retinopathy of prematurity. Surviving infants, including the subset born at less than 28 weeks gestation, have been assessed at a home visit. Parents completed a comprehensive questionnaire and children underwent a visual assessment and were tested on the Wechsler Intelligence Scale for Children. RESULTS: Of 126 liveborn infants less than 28 weeks gestation, 80 (64%) survived to 7-8 years. Sixty eight children (97% survivors resident in New Zealand) were assessed: 72% had no, and 86% no or only mild disability, 77% had some visual problem, with close to one third having myopia, strabismus or requiring spectacles and 32% received Ministry of Education funded special needs assistance. CONCLUSIONS: There have been few long-term follow-up studies of infants of less than 28 weeks gestation born in a defined geographical area. The outcome for New Zealand infants is comparable with that in other published data. PMID- 9734527 TI - Trends in motorcyclist and occupant fatalities and serious injuries due to traffic crashes. AB - AIM: To describe trends in motorcycle traffic crashes and compare these trends in other crashes and vehicle registrations. METHOD: National fatality and public hospital inpatient data were used to select cases. Relative changes in both deaths and hospitalisations, and vehicle registrations were examined in each year during the period 1980-98. RESULTS: The results show that deaths and serious injuries to motorcyclists have declined substantially over the period 1980-1995. The trend in serious injuries to motorcyclists closely followed the trend in motorcycle registrations. The association was less evident for motorcyclist fatalities and for occupants there was no clear association with trends in registrations for either outcome. CONCLUSIONS: The reduction in motorcycle injuries has contributed substantially to our improved road safety record. We need to protect these gains but in order to do this we need a clear understanding of how they were achieved. Such an understanding is currently lacking. PMID- 9734528 TI - The impact of acne: a study of adolescents' attitudes, perception and knowledge. AB - AIM: To assess adolescent students' attitudes to, perceptions and knowledge of acne and to assess the effect of acne on daily living. METHOD: Students from Auckland sixth and seventh form classes were selected from ten Auckland secondary schools using a randomisation process which ensured proportional representation by socioeconomic group and gender. Eight hundred and forty-seven students completed a written questionnaire on the subject of acne vulgaris and had their acne examined. Their acne was graded using a modification of the Leeds system which determines severity on the basis of number, extent and nature of the skin lesions. RESULTS: Acne was present in 91% of males and 79% of females. Students' perceptions of the severity of their acne were significantly related to objective clinical assessment (p=0.00001). Severity of acne determined the extent of embarrassment (p<0.00001) and the lack of enjoyment of and participation in social activities (p<0.00002). These analyses were significant for both males and females. Students had misconceptions regarding the causes of acne. Parental occupation and ethnic group were related to knowledge of treatment for acne. CONCLUSION: Acne causes personal and social difficulties for a large number of adolescent students. There is a need for all students to have access to appropriate information and health services so that the social and psychological consequences of acne are minimised. PMID- 9734529 TI - The Otago Medical School revisited. PMID- 9734530 TI - Chickenpox immunisation in New Zealand. AB - PREVENTION: The appropriate use of varicella vaccine, effective in the prevention of chickenpox, has been considered by a Ministry of Health Working Party in 1996 and 1997, including discussion at a workshop held in Wellington, 26-27 June 1996. The introduction of varicella vaccine into the routine childhood immunisation schedule was not supported at this stage. The use of the only varicella vaccine for which the Minister of Health has given consent for distribution in New Zealand, Varilrix (SmithKline Beecham Limited), in healthy children aged nine months to 13 years inclusive, was supported. Consent has not been given for the use of Varilrix in immunocompromised people or in adults. This report discusses other groups that could be candidates for vaccination, such as children with deteriorating renal function and susceptible health care workers who regularly come into contact with especially vulnerable patients. In these cases, the vaccine would need to be administered on a named patient basis. The use of Varilrix in immunocompromised people was not supported. SURVEILLANCE: Enhanced surveillance of chickenpox and zoster are required in New Zealand. Adverse reactions to Varilrix should be carefully monitored. OUTBREAK CONTROL: There are insufficient data at present to support the use of Varilrix in outbreak control. The frequency, cost and current management of nosocomial outbreaks should be ascertained. This information may also assist in the decision whether to incorporate a varicella vaccine into the routine childhood immunisation schedule in the future. PMID- 9734532 TI - The caduceus--a further interpretation. PMID- 9734531 TI - Antimicrobial susceptibility of Campylobacter and Yersinia enterocolitica isolates. PMID- 9734533 TI - Physical assault in New Zealand. PMID- 9734534 TI - Panic and phobic anxiety: phenotypes, endophenotypes, and genotypes. PMID- 9734535 TI - Images in neuroscience. Brain development, VI: radial migration and cortical evolution. PMID- 9734537 TI - Vulnerability to posttraumatic stress disorder in adult offspring of Holocaust survivors. AB - OBJECTIVE: There has been considerable controversy regarding the impact of the Holocaust on the second generation, but few empirical data are available that systematically document trauma exposure and psychiatric disorder in these individuals. To obtain such data, the authors examined the prevalence of stress and exposure to trauma, current and lifetime posttraumatic stress disorder (PTSD), and other psychiatric diagnoses in a group of adult offspring of Holocaust survivors (N=100) and a demographically similar comparison group (N=44). METHOD: Subjects were recruited from both community and clinical populations and were evaluated with the use of structured clinical instruments. Stress and trauma history were evaluated with the Antonovsky Life Crises Scale and the Trauma History Questionnaire, PTSD was diagnosed with the Clinician Administered PTSD Scale, and other psychiatric disorders were evaluated according to the Structured Clinical Interview for DSM-IV. RESULTS: The data show that although adult offspring of Holocaust survivors did not experience more traumatic events, they had a greater prevalence of current and lifetime PTSD and other psychiatric diagnoses than the demographically similar comparison subjects. This was true in both community and clinical subjects. CONCLUSIONS: The findings demonstrate an increased vulnerability to PTSD and other psychiatric disorders among offspring of Holocaust survivors, thus identifying adult offspring as a possible high-risk group within which to explore the individual differences that constitute risk factors for PTSD. PMID- 9734536 TI - Panic and phobic anxiety: defining phenotypes for genetic studies. AB - OBJECTIVE: With recent advances in molecular genetics, the rate-limiting step in identifying susceptibility genes for psychiatric disorders has become phenotype definition. The success of psychiatric genetics may require the development of a "genetic nosology" that can classify individuals in terms of the heritable aspects of psychopathology. The authors' aim is to begin to apply this analysis to the anxiety disorders, focusing on panic and phobic disorders. METHOD: Two parallel traditions of defining anxiety phenotypes are reviewed: the first, more closely identified with clinical psychiatry, has identified categorical diagnoses (e.g., panic disorder and social phobia). The other, more closely identified with psychological studies of personality development, has examined dimensional traits (e.g., neuroticism) and anxious temperament (e.g., behavioral inhibition). RESULTS: The authors suggest that a genetic nosology of panic and phobic disorders may incorporate features of both traditions and discuss strategies for optimizing genetic approaches to anxiety including 1) studying phenotypic extremes, 2) identifying biological trait markers, and 3) using animal models to identify candidate loci. CONCLUSIONS: An important dividend from the effort to define the boundaries of heritable phenotypes for genetic studies of anxiety may be a refinement of the nosology of anxiety disorders. PMID- 9734538 TI - Efficacy of Psychoeducational Group Therapy in reducing symptoms of posttraumatic stress disorder among multiply traumatized women. AB - OBJECTIVE: The role of group therapy in treatment of posttraumatic stress disorder (PTSD) has been traditionally restricted to issues of self-esteem and interpersonal relationships, rather than primary symptoms of the disorder. In this study, the authors examined the effectiveness of a 16-week trauma-focused, cognitive-behavioral group therapy, named Interactive Psychoeducational Group Therapy, in reducing primary symptoms of PTSD in five groups (N=29) of multiply traumatized women diagnosed with chronic PTSD. METHOD: The authors made assessments at baseline, at 1-month intervals during treatment, at termination, and at 6-month follow-up by using self-report and structured interview measures of PTSD and psychiatric symptoms. The absence of a control group limits the conclusions drawn from the study. RESULTS: At termination, subjects showed significant reductions in all three clusters of PTSD symptoms (i.e., reexperiencing, avoidance, and hyperarousal) and in depressive symptoms; they showed near-significant reductions in general psychiatric and dissociative symptoms, at termination. These improvements were sustained at 6-month follow-up. CONCLUSIONS: The role of group therapy in PTSD treatment should not be prematurely restricted to addressing self-esteem and interpersonal dimensions only. The use of structured, cognitive-behavioral elements within the group format may allow for more targeted treatment of core symptoms of the disorder. PMID- 9734539 TI - Cerebral glucose metabolism in women with panic disorder. AB - OBJECTIVE: This study was undertaken to clarify earlier inconsistent findings in brain metabolic topography in panic disorder patients at rest. METHOD: Positron emission tomography (PET) with [18F]fluorodeoxyglucose was used to determine cerebral metabolic activity in six female patients with a DSM-III-R diagnosis of panic disorder and in six healthy female volunteers. All patients with panic disorder were medication free and were sensitive to lactate infusion. RESULTS: A significant increase in glucose metabolism was found in the left hippocampus and parahippocampal area of the panic disorder subjects in comparison with that found in the healthy subjects. In addition, a significant decrease in metabolism was found in the right inferior parietal and right superior temporal brain regions of the panic disorder subjects in comparison with that of the normal subjects. There was no significant correlation between scores for the severity of panic disorder or for the severity of lactate-induced panic attack and the quantified PET abnormality. CONCLUSIONS: These data provide further support for the hypothesis of an abnormal brain metabolism in the hippocampal and parahippocampal area in individuals with panic disorder and also suggest other areas of aberrant brain metabolism in this disorder. PMID- 9734540 TI - CO2-induced panic attacks: a twin study. AB - OBJECTIVE: The authors investigated the role of genetic factors in 35% CO2 induced panic attacks. METHOD: Ninety twins recruited from the general population were challenged with one-vital-capacity inhalations of 35% CO2-65% O2. Probandwise concordance rates were calculated and rates compared for monozygotic and for dizygotic twins. RESULTS: A significantly higher concordance was found for 35% CO2-induced panic attacks among monozygotic than dizygotic twins (55.6% versus 12.5%). CONCLUSIONS: These results suggest a relevant role of genetic factors in 35% CO2-induced panic attacks. PMID- 9734541 TI - Sertraline in the treatment of panic disorder: a double-blind multicenter trial. AB - OBJECTIVE: This study determined the efficacy and safety of sertraline in the treatment of patients with panic disorder. METHOD: The study was a randomized, double-blind, parallel-group, flexible-dose comparison of sertraline and placebo in outpatients with a DSM-III-R diagnosis of panic disorder with or without agoraphobia. After a 2-week single-blind placebo lead-in, 168 patients entered a 10-week double-blind phase in which they were randomly assigned to treatment with either sertraline or placebo. RESULTS: Sertraline was significantly more effective than placebo in decreasing the number of full and limited-symptom panic attacks. Among patients who completed the study, the mean number of panic attacks per week dropped by 88% in the sertraline-treated patients and 53% in the placebo treated patients. Sertraline-treated patients also had significantly more improvement than placebo-treated patients in scores on the Quality of Life Enjoyment and Satisfaction Questionnaire, patient global evaluation, and Clinical Global Impression severity of illness and global improvement scales. Overall, patients tolerated sertraline well, and only 9% terminated treatment because of side effects. CONCLUSIONS: Sertraline is an effective and well-tolerated treatment for patients with panic disorder. PMID- 9734542 TI - Two-year outcome in first-episode schizophrenia: predictive value of symptoms for quality of life. AB - OBJECTIVE: Many studies have validated the grouping of schizophrenic symptoms into three independent dimensions: negative, psychotic, and disorganized. Negative symptoms are considered to be an important prognostic indicator, but this clinical observation requires further empirical study, especially with respect to psychosocial functioning. When present at the onset of the first episode, negative symptoms suggest that the patient will develop significant psychosocial impairment. The predictive values of the psychotic and disorganized symptom dimensions, on the other hand, have been less certain. METHOD: In this study of 50 first-episode schizophrenic patients, who were mostly neuroleptic naive at intake, the authors examined the relationship between the severity of these three symptom dimensions (measured by using the Scale for the Assessment of Negative Symptoms and the Scale for the Assessment of Positive Symptoms) at index hospitalization and quality of life at 2-year follow-up. RESULTS: Negative symptom severity was positively and significantly correlated with later occupational impairment, financial dependence on others, impaired relationships with friends, impaired ability to enjoy recreational activities, and global assessment of functioning. The magnitudes of correlation between the levels of psychotic symptoms or disorganized symptoms and 2-year quality of life measures were comparatively lower. Analyses using multivariate regression statistics also revealed similar findings. CONCLUSIONS: Severity of negative symptoms at index hospitalization may be a portent of poor outcome. In general, severity of psychotic or disorganized symptoms at intake does not appear to predict subsequent quality of life. PMID- 9734543 TI - Spontaneous abnormal involuntary movements in first-episode schizophrenia and schizophreniform disorder: baseline rate in a group of patients from an Irish catchment area. AB - OBJECTIVE: This study investigated the rate of spontaneous abnormal involuntary movements in a group of patients presenting with a first episode of schizophrenia or schizophreniform psychosis. METHOD: Seventy-nine patients with a first episode of schizophrenia or schizophreniform psychosis who presented to a catchment area psychiatric service over a 3-year period, and who were neuroleptic-naive or had been medicated for less than 1 month, were examined for the presence of involuntary movements with use of the Abnormal Involuntary Movement Scale. RESULTS: Six patients (7.6%) had spontaneous dyskinesia as defined by the criteria of Schooler and Kane, and nine other patients had mild orofacial involuntary movements. The patients with spontaneous dyskinesia had completed significantly fewer years of education than the patients without dyskinesia. Spontaneous involuntary movements were unrelated to age at presentation for treatment. CONCLUSIONS: Spontaneous abnormal involuntary movements were evident among a proportion of patients with first-episode schizophrenia or schizophreniform psychosis at baseline presentation and were associated with reduced educational attainment. This finding supports previous suggestions that abnormal involuntary movements in schizophrenia may be related to the pathophysiology of the illness and therefore cannot be attributed entirely to the adverse effects of neuroleptic medication. PMID- 9734544 TI - Markers of glutamatergic neurotransmission and oxidative stress associated with tardive dyskinesia. AB - OBJECTIVE: Tardive dyskinesia is a movement disorder affecting 20%-40% of patients treated chronically with neuroleptic drugs. The dopamine supersensitivity hypothesis cannot account for the time course of tardive dyskinesia or for the persistence of tardive dyskinesia and the associated structural changes after neuroleptics are discontinued. The authors hypothesized that neuroleptics enhance striatal glutamatergic neurotransmission by blocking presynaptic dopamine receptors, which causes neuronal damage as a consequence of oxidative stress. METHOD: CSF was obtained from 20 patients with schizophrenia, 11 of whom had tardive dyskinesia. Markers for oxidative stress, including superoxide dismutase, lipid hydroperoxide, and protein carbonyl groups, and markers for excitatory neurotransmission, including N-acetylaspartate, N acetylaspartylglutamate, aspartate, and glutamate, were measured in the CSF specimens. Patients were also rated for tardive dyskinesia symptoms with the Abnormal Involuntary Movement Scale. RESULTS: Tardive dyskinesia patients had significantly higher concentrations of N-acetylaspartate, N acetylaspartylglutamate, and aspartate in their CSF than patients without tardive dyskinesia when age and neuroleptic dose were controlled for. The significance of the higher levels of protein-oxidized products associated with tardive dyskinesia did not pass Bonferroni correction, however. Tardive dyskinesia symptoms correlated positively with markers of excitatory neurotransmission and protein carbonyl group and negatively with CSF superoxide dismutase activity. CONCLUSIONS: These findings suggest that there are elevated levels of oxidative stress and glutamatergic neurotransmission in tardive dyskinesia, both of which may be relevant to the pathophysiology of tardive dyskinesia. PMID- 9734545 TI - Sustained attention deficit and schizotypal personality features in nonpsychotic relatives of schizophrenic patients. AB - OBJECTIVE: The authors investigated whether nonpsychotic relatives of schizophrenic probands have an elevated risk of deficits in sustained attention as measured by the Continuous Performance Test (CPT), whether such deficits are associated with specific factors of schizotypy, and whether poor CPT performance by probands predicts poor performance by their relatives. In addition, the heritability of CPT performance in the families of schizophrenic probands was estimated. METHOD: The study subjects were 60 schizophrenic probands, 148 of their first-degree relatives, 20 normal comparison probands, and 42 of the comparison probands' first-degree relatives. Subjects completed undegraded and 25% degraded sessions of the CPT and were interviewed with use of the Chinese version of the Diagnostic Interview for Genetic Studies. Subjects' CPT sensitivity indexes, d', were standardized against those of a community sample of 345 subjects, with adjustment for age, sex, and level of education. RESULTS: On average, the d' values of the relatives of schizophrenic probands were lower than those of the relatives of comparison probands but higher than those of schizophrenic probands. Lower sensitivity indexes among the relatives of schizophrenic patients were associated with the interpersonal dysfunction and disorganization factors of schizotypy but not the cognitive/perceptual factor. When schizophrenic probands were divided into two subgroups by a cutoff of -3.0 for adjusted z score on the CPT, the d' values of relatives of probands with CPT deficits were lower than those of relatives of probands without deficits. The estimated heritability of performance on the CPT ranged from 0.48 to 0.62. CONCLUSIONS: Sustained attention deficit may be a genetic vulnerability marker for schizophrenia, and it may be more useful in linkage analysis than traditional phenotype definitions of schizophrenia. PMID- 9734546 TI - Summer birth and the deficit syndrome of schizophrenia. AB - OBJECTIVE: Patients with the deficit syndrome differ from other patients with schizophrenia relative to physiological correlates, course of illness, and response to treatment. Because of the abnormal seasonality of birth among persons with schizophrenia, the authors examined the relation between this risk factor and the deficit syndrome. METHOD: Findings in two clinical groups suggested an increase in summer births among deficit syndrome patients. The association between summer birth and the deficit syndrome was then examined in a catchment area study of first-admission patients with psychosis. RESULTS: In the catchment area sample, summer birth was also significantly associated with the deficit syndrome; negative symptoms broadly defined were not. CONCLUSIONS: These findings add to the increasing evidence that 1) patients with the deficit syndrome have a disease with an etiopathophysiology separate from that of other patients with what is now called schizophrenia and 2) the correlates of broadly defined negative symptoms are different from those for the deficit syndrome. The previously reported association between winter birth and schizophrenia appears to apply to nondeficit schizophrenia only. PMID- 9734547 TI - Attributional style and depression in pregnant teenagers. AB - OBJECTIVE: Study 1 evaluated whether pregnancy is a stressful life event for teenagers and is associated with depression for teenagers with a pessimistic attributional style but not for those with an optimistic attributional style. Study 2 replicated unexpected findings from study 1. Study 3 evaluated whether pregnant teenagers with a pessimistic attributional style will be more depressed after childbirth than optimistic pregnant teenagers. METHOD: In study 1, 122 pregnant and 96 nonpregnant teenagers completed questionnaires assessing depression and attributional style. In study 2, 43 pregnant teenagers and 49 nonpregnant teenagers completed the same questionnaires. In study 3, subjects from studies 1 and 2 were contacted by mail and asked to complete the same questionnaires; 63 (38%) did so. RESULTS: In studies 1 and 2, pregnant teenagers with a pessimistic attributional style were less depressed than those with an optimistic attributional style and the nonpregnant group. In study 3, pessimistic teenagers experienced a higher level of depression than nonpessimistic adolescents following childbirth. CONCLUSIONS: Pregnancy may serve to protect pessimistic teenagers from experiencing depression. The subsequent demands of motherhood may remove any protection from depression afforded by the pregnancy. The experience of depression being relieved by another pregnancy may be a previously unidentified risk factor for repeated pregnancies in teenagers. PMID- 9734548 TI - Longitudinal population-based twin study of retrospectively reported premenstrual symptoms and lifetime major depression. AB - OBJECTIVE: While family and twin studies suggest that retrospectively reported premenstrual symptoms are heritable, these studies have not accounted for the unreliability of such measures. In addition, we know little about the relationship of the familial risk factors for premenstrual symptoms and major depression. METHOD: Lifetime major depression and premenstrual-related tiredness, sadness, and irritability were assessed twice over 6 years in 1,312 menstruating female twins ascertained from a population-based twin register. A twin measurement model--which permits estimation of the etiologic roles of genetic and environmental factors with correction for errors of measurement or short-term temporal fluctuations--was applied to these data. RESULTS: A single premenstrual symptom factor was found that was moderately stable over time. The best-fitting twin-measurement model estimated the heritability of the stable component of premenstrual symptoms at 56% and showed no impact of family-environmental factors. A bivariate twin-measurement model estimated that the genetic and environmental risk factors for lifetime major depression contributed only modestly to the etiology of premenstrual syndrome. No evidence was found for significant biases in the twin method. CONCLUSIONS: Retrospectively reported premenstrual-related symptoms of depression and anxiety are moderately stable over time and, when correction is made for this level of stability, substantially heritable. The genetic and environmental risk factors for these premenstrual symptoms and lifetime major depression are not closely related. PMID- 9734549 TI - No effect of depression on [(15)O]H2O PET response to intravenous d-fenfluramine. AB - OBJECTIVE: Subnormal prolactin responses to the serotonin-releasing agonist fenfluramine occur in depression. Since many measures of serotonin pathology occur in depression, abnormal responses to fenfluramine may occur in brain structures other than the hypothalamic-pituitary axis. One study compared six depressed and six healthy subjects' responses to oral d,l-fenfluramine by assessing [18F]fluorodeoxyglucose uptake as detected by positron emission tomography (PET). That study showed several abnormalities within the cortex, and the authors concluded that low responsivity to d,l-fenfluramine is widespread in depression. In this study abnormalities in regional neuromodulation by serotonin in major depression were assessed with intravenous d-fenfluramine and [(15)O]H2O PET. METHOD: Changes in regional cerebral blood flow (CBF) were detected by using [(15)O]H2O PET after administration of intravenous d-fenfluramine to 13 depressed and 18 healthy women. The PET scans were done 20 and 5 minutes before and 20 and 35 minutes after d-fenfluramine administration. Differences between the depressed and healthy groups in change in regional CBF (mean postfenfluramine minus mean prefenfluramine) were analyzed by using statistical parametric mapping. RESULTS: There were no significant differences between depressed and healthy subjects; in fact, changes in regional CBF after intravenous d-fenfluramine were remarkably similar. CONCLUSIONS: Degrees of neuronal responsivity to d-fenfluramine are similar in depressed and healthy subjects. Differences between the previous and current findings may be accounted for by greater specificity of intravenous d fenfluramine to serotonin release, timing of scans, paucity of suicidal subjects in the current study, or greater variance in regional CBF from direct vascular effects of serotonin. PMID- 9734550 TI - Optimal length of continuation therapy in depression: a prospective assessment during long-term fluoxetine treatment. AB - OBJECTIVE: The purpose of this study was to determine prospectively the optimal length of therapy in a long-term, placebo-controlled continuation study of patients who responded to acute fluoxetine treatment for major depression (defined by DSM-III-R). METHOD: The study was conducted at five outpatient psychiatric clinics in the United States. Patients who met criteria for remission after 12 or 14 weeks of open-label acute fluoxetine therapy, 20 mg/day (N=395 of 839 patients), were randomly assigned to one of four arms of a double-blind treatment study (50 weeks of placebo, 14 weeks of fluoxetine and then 36 weeks of placebo, 38 weeks of fluoxetine and then 12 weeks of placebo, or 50 weeks of fluoxetine). Relapse rate was the primary outcome measure. Both Kaplan-Meier estimates and observed relapse rates were assessed in three fixed 12-week intervals after double-blind transfers from fluoxetine to placebo at the start of the double-blind period and after 14 and 38 weeks of continued fluoxetine treatment. RESULTS: Relapse rates (Kaplan-Meier estimates) were lower among the patients who continued to take fluoxetine compared with those transferred to placebo in both the first interval, after 24 total weeks of treatment (fluoxetine, 26.4%; placebo, 48.6%), and the second interval, after 38 total weeks of treatment (fluoxetine, 9.0%; placebo, 23.2%). In the third interval, after 62 total weeks of treatment, rates were not significantly different between the groups (fluoxetine, 10.7%; placebo, 16.2%). CONCLUSIONS: Patients treated with fluoxetine for 12 weeks whose depressive symptoms remit should continue treatment with fluoxetine for at least an additional 26 weeks to minimize the risk of relapse. PMID- 9734551 TI - Factual sources of psychiatric patients' perceptions of coercion in the hospital admission process. AB - OBJECTIVE: The purpose of this study was to determine what predicts patients' perceptions of coercion surrounding admission to a psychiatric hospital. METHOD: For 171 cases, the authors integrated data from interviews with patients, admitting clinicians, and other individuals involved in the patients' psychiatric admissions with data from the medical records. Using a structured set of procedures, coders determined whether or not nine coercion-related behaviors occurred around the time of admission. Correlation and regression analyses were used to describe the predictors of patients' scores on the MacArthur Perceived Coercion Scale. RESULTS: The use of legal force, being given orders, threats, and "a show of force" were all strongly correlated with perceived coercion. A least squares regression accounted for 43.3% of the variance in perceived coercion. The evidence also suggested that force is typically only used in conjunction with less coercive pressures. CONCLUSIONS: Force and negative symbolic pressures, such as threats and giving orders about admission decisions, induce perceptions of coercion in persons with mental illness. Positive symbolic pressures, such as persuasion, do not induce perceptions of coercion. Such positive pressures should be tried in order to encourage admission before force or negative pressures are used. PMID- 9734552 TI - A bibliography of mental patients' autobiographies: an update and classification system. AB - OBJECTIVE: This article brings to the present earlier bibliographies of books written by former mental patients. These books provide an inside view of mental disorder that can be useful in teaching, public education, theory, and research, and they have played a catalyzing role in mental health reform and in theory development. METHOD: The authors list seven anthologies and 48 autobiographies of former patients published since 1980 and introduce a classification system intended to increase the research value of this important archive. RESULTS: Recent books of this genre show more individuals with a mood disorder and more therapists and more women as authors. CONCLUSIONS: The research potential of these books suggests the value of an electronic database for classifying and retrieving the information they contain. PMID- 9734553 TI - Prader-Willi syndrome. PMID- 9734554 TI - Lack of association between neuroleptic malignant syndrome and polymorphisms in the 5-HT1A and 5-HT2A receptor genes. AB - OBJECTIVE: The molecular basis of neuroleptic malignant syndrome is unclear, but studies suggest that genetic factors are involved in its pathogenesis. Considering possible involvement of the serotonergic system in neuroleptic malignant syndrome, the authors examined the association between neuroleptic malignant syndrome and polymorphisms of the 5-HT1A and 5-HT2A receptor genes. METHOD: The authors examined the frequencies of gene polymorphisms in the 5-HT1A (Arg219Leu) and 5-HT2A (Thr25Asn and His452Tyr) receptor genes in 29 patients previously diagnosed with neuroleptic malignant syndrome, 94 neuroleptic-treated patients with schizophrenia who had no history of neuroleptic malignant syndrome, and 94 healthy comparison subjects. Polymerase chain reaction and restriction fragment length polymorphism analyses were used to screen gene mutations. RESULTS: No polymorphic allele was detected in the patients who had experienced the neuroleptic malignant syndrome. CONCLUSIONS: The authors cannot conclude that polymorphisms in the 5-HT1A and 5HT2A receptor genes are factors determining susceptibility to the neuroleptic malignant syndrome. PMID- 9734555 TI - Pilot study of the cytochrome P450-2D6 genotype in a psychiatric state hospital. AB - OBJECTIVE: The authors conducted a pilot study to develop preliminary data on the frequency of cytochrome P450-2D6 (CYP2D6) genotypes in state psychiatric hospital patients and to establish population sizes needed to determine potential clinical relevance in therapeutic outcome. METHOD: One hundred consecutive inpatients at Eastern State Hospital in Kentucky who provided informed consent were genotyped at the CYP2D6 locus during their hospital stay. RESULTS: Twelve of the patients were CYP2D6 deficient, and four carried the *1Xn or *2Xn allele associated with ultrarapid metabolism; all of these patients were Caucasian (N=87). The rate of deficiency in CYP2D6 expression in these Caucasian state psychiatric hospital patients (14%) was twice that of the U.S. population (7%). The patients with CYP2D6 deficiency also appeared more likely to experience side effects in response to CYP2D6 medications. CONCLUSIONS: This study, limited by a small number of subjects, suggests that one-fifth of Caucasians admitted to a state hospital in Kentucky had genotypes associated with extremes in CYP2D6 activity that may have affected their response to CYP2D6 medications. PMID- 9734556 TI - Auditory mismatch negativity in schizophrenia: topographic evaluation with a high density recording montage. AB - OBJECTIVE: The mismatch negativity, a negative component in the auditory event related potential, is thought to index automatic processes involved in sensory or echoic memory. The authors' goal in this study was to examine the topography of auditory mismatch negativity in schizophrenia with a high-density, 64-channel recording montage. METHOD: Mismatch negativity topography was evaluated in 23 right-handed male patients with schizophrenia who were receiving medication and in 23 nonschizophrenic comparison subjects who were matched in age, handedness, and parental socioeconomic status. The Positive and Negative Syndrome Scale was used to measure psychiatric symptoms. RESULTS: Mismatch negativity amplitude was reduced in the patients with schizophrenia. They showed a greater left-less-than right asymmetry than comparison subjects at homotopic electrode pairs near the parietotemporal junction. There were correlations between mismatch negativity amplitude and hallucinations at left frontal electrodes and between mismatch negativity amplitude and passive-apathetic social withdrawal at left and right frontal electrodes. CONCLUSIONS: Mismatch negativity was reduced in schizophrenia, especially in the left hemisphere. This finding is consistent with abnormalities of primary or adjacent auditory cortex involved in auditory sensory or echoic memory. PMID- 9734557 TI - Functional hypofrontality and working memory dysfunction in schizophrenia. AB - OBJECTIVE: Hypofrontality is a common but not invariable finding in schizophrenia. Inconsistencies in the literature may reflect, in part, the fact that abnormal physiological responses in the prefrontal cortex are best identified under conditions that place well-specified functional demands on this region. METHOD: The authors studied eight patients with schizophrenia and eight matched comparison subjects using [(15)O]H2O positron emission tomography and the "N-back" task, which activates the prefrontal cortex as a function of working memory load in normal subjects. RESULTS: Under low-working-memory-load conditions, the accuracy of both groups in the N-back task was equal, but when the memory load increased, the patients' performance deteriorated more than did that of the comparison subjects. The regional cerebral blood flow response to increased working memory load was significantly reduced in the patients' right dorsolateral prefrontal cortex. CONCLUSIONS: These results confirm the importance of using tasks that tap specific cognitive functions, linked to specific neural systems, in studies of brain-behavior relationships in schizophrenia. Hypofrontality is reliably demonstrated in schizophrenia during tasks that engage working memory functions of the prefrontal cortex. PMID- 9734558 TI - Reduced Purkinje cell size in the cerebellar vermis of elderly patients with schizophrenia. AB - OBJECTIVE: The authors' goal was to compare the size and linear density of Purkinje cells in the cerebellar vermis of subjects with and without schizophrenia. METHOD: Blocks of alcohol-fixed cerebellar vermis were dissected at autopsy from the brains of 14 elderly patients with schizophrenia and 13 elderly subjects with no history of neuropsychiatric illness. The blocks of vermis were sectioned and stained with 1% cresyl violet. The linear density and cross-sectional area of Purkinje cells were measured by using computer-assisted image analysis. The subjects with schizophrenia had been assessed with clinical rating scales within 1 year prior to death. RESULTS: The average cross-sectional areas of Purkinje cells of the patients with schizophrenia were significantly smaller (by 8.3%) than those of the subjects without neuropsychiatric illness. No difference in Purkinje cell linear density was observed between the two groups. Significant correlations were seen between Purkinje cell size and scores on the Mini-Mental State, the Brief Psychiatric Rating Scale, and the antipsychotic drug dose. CONCLUSIONS: These data indicate cerebellar involvement in schizophrenia; they are also consistent with reports of reduced neuronal size in other brain regions of patients with schizophrenia. These findings support a model of wide spread central nervous system abnormality in schizophrenia. PMID- 9734559 TI - Treatment of aggression in schizophrenia. PMID- 9734560 TI - Preventing contractures [correction of contractions] in neuroleptic malignant syndrome and dystonia. PMID- 9734561 TI - Provocation of a posttraumatic flashback by cholecystokinin tetrapeptide? PMID- 9734562 TI - Olanzapine augmentation of fluoxetine in the treatment of trichotillomania. PMID- 9734563 TI - Consistency of memory among veterans of Operation Desert Storm. PMID- 9734564 TI - Consistency of memory among veterans of Operation Desert Storm. PMID- 9734565 TI - APA Practice Guideline for schizophrenia: risperidone equivalents. American Psychiatric Association. PMID- 9734566 TI - Sertindole versus haloperidol for schizophrenia. PMID- 9734567 TI - Sertindole versus haloperidol for schizophrenia. PMID- 9734568 TI - Treatment outcome for dissociative identity disorder. PMID- 9734569 TI - Comments on abortion. PMID- 9734570 TI - Comments on diagnosis including symptoms of turbulent grief. PMID- 9734571 TI - Outcome measures of an Australian breast-screening program. PMID- 9734572 TI - Use of complementary medicines: scientific and ethical issues. PMID- 9734573 TI - Spina bifida. PMID- 9734574 TI - Interval breast cancers in an Australian mammographic screening program. AB - OBJECTIVE: To determine the incidence of interval cancers which occurred in the first 12 months after mammographic screening at a mammographic screening service. DESIGN: Retrospective analysis of data obtained by crossmatching the screening Service and the New South Wales Central Cancer Registry databases. SETTING: The Central & Eastern Sydney Service of BreastScreen NSW. PARTICIPANTS: Women aged 40 69 years at first screen, who attended for their first or second screen between 1 March 1988 and 31 December 1992. MAIN OUTCOME MEASURES: Interval-cancer rates per 10000 screens and as a proportion of the underlying incidence of breast cancer (as estimated by the underlying rate in the total NSW population). RESULTS: The 12-month interval-cancer incidence per 10000 screens was 4.17 for the 40-49 years age group (95% confidence interval [CI], 1.35-9.73) and 4.64 for the 50-69 years age group (95% CI, 2.47-7.94). Proportional incidence rates were 30.1% for the 40 49 years age group (95% CI, 9.8-70.3) and 22% for the 50-69 years age group (95% CI, 11.7-37.7). There was no significant difference between the proportional incidence rate for the 50-69 years age group for the Central & Eastern Sydney Service and those of major successful overseas screening trials. CONCLUSION: Screening quality was acceptable and should result in a significant mortality reduction in the screened population. Given the small number of cancers involved, comparison of interval-cancer statistics of mammographic screening programs with trials requires age-specific or age-adjusted data, and consideration of confidence intervals of both program and trial data. PMID- 9734575 TI - Helicobacter pylori infection: an added stressor on iron status of women in the community. AB - OBJECTIVE: To explore a possible association between Helicobacter pylori infection and iron status. DESIGN: Cross-sectional study. SETTING: Ballarat (a major regional city in Victoria), population 78000, October November 1997. PARTICIPANTS: 160 women and 152 men, a subsample of participants in a cardiovascular disease risk factor prevalence survey for whom frozen plasma was available. MAIN OUTCOME MEASURES: H. pylori IgG antibody status by enzyme immunoassay; iron intake; plasma iron, transferrin and ferritin concentrations. RESULTS: 28% of women and 33% of men were infected with H. pylori. The mean (SEM) plasma ferritin concentration of infected women (59.3 [7.6] microg/L) was significantly lower than for non-infected women (88.8 [7.9] microg/L; P=0.002), after adjusting for age. Mean daily dietary iron intakes were similar in infected and non-infected women. CONCLUSIONS: H. pylori infection appears to be an additional stressor on women's iron status, but the mechanism remains to be determined. PMID- 9734577 TI - Acupuncture in Australian general practice: practitioner characteristics. AB - OBJECTIVES: To ascertain the extent of the use of acupuncture and the characteristics of general practitioners using acupuncture. DESIGN: Secondary analysis of 1996 Health Insurance Commission data on claims by all non-specialist medical practitioners for Medicare Benefits Schedule items for an attendance where acupuncture was performed by a medical practitioner. MAIN OUTCOME MEASURES: Use of acupuncture by general practitioners and the practitioners' sex, age, place of primary medical qualification, and the socioeconomic disadvantage index of the practitioners' practice. RESULTS: 15.1% of general practitioners claimed for acupuncture. Acupuncture was more likely to be provided by male practitioners, by those aged 35-54 years, and by practitioners who have an overseas primary medical qualification. The socioeconomic index of the practice did not significantly affect the number of claims for acupuncture. CONCLUSION: Acupuncture is used by about one in seven general practitioners. Its use is associated with middle-aged practitioners, who presumably have more clinical experience. This level of use by experienced doctors suggests that a critical review of the appropriate role of acupuncture in general practice should be considered. PMID- 9734576 TI - Psychological morbidity and quality of life in Australian women with early-stage breast cancer: a cross-sectional survey. AB - OBJECTIVE: To determine the prevalence of psychological morbidity and describe quality of life in women with early-stage breast cancer. DESIGN: Cross-sectional descriptive study (3 months after conservative breast surgery or mastectomy) of patients from nine general hospitals in Melbourne, Victoria, October 1994 to March 1997. PARTICIPANTS: 303 women with early-stage breast cancer entering a randomised trial of adjuvant psychological group therapy; mean age, 46 years (SD, 8). MAIN OUTCOME MEASURES: Diagnostic and Statistical Manual of Mental Health (DSM)-IV psychiatric diagnoses generated by the Monash Interview for Liaison Psychiatry; quality-of-life data based on the the European Organization for Research and Treatment of Cancer quality-of-life questionnaire (QLQ)-C30 (core) and QLQ-BR23 (breast module) instruments. RESULTS: 45% of the women (135/303) had a psychiatric disorder; 42% (127) of the sample had depression or anxiety, or both; there was minor depression in 82 (27.1%), an anxiety disorder in 26 (8.6%), major depression in 29 (9.6%) and a phobic disorder in 21 (6.9%). 20% of women (61) had more than one disorder. On quality-of-life measures nearly one-third of the women felt less attractive and most had lost interest in sexual activity. There was substantial distress about hair loss. Symptoms of lymphoedema were described by 13 women (4.3%). Breast conservation surgery was associated with a better body image (P<0.01). CONCLUSION: Women recently diagnosed with early-stage breast cancer have high rates of psychiatric and psychological disturbance. Quality of life is substantially affected. Clinicians should actively explore their patients' psychological adjustment to enable early recognition and treatment of these disorders. PMID- 9734578 TI - Cerebral histoplasmosis in an Australian patient with systemic lupus erythematosus. AB - A 39-year-old woman with systemic lupus erythematosus suffered a prolonged neurological illness associated with very low levels of glucose in her cerebrospinal fluid (CSF). Six months later, and after numerous CSF investigations, Histoplasma capsulatum was cultured. To our knowledge, this is the first report of cerebral histoplasmosis in Australia in a patient who is not HIV positive. PMID- 9734579 TI - The diastolic debate: is it time to discard Korotkoff phase IV in favour of phase V for blood pressure measurements in pregnancy? AB - Current guidelines recommend using Korotkoff phase IV for measuring diastolic blood pressure in pregnant women. However, phase IV does not approximate "true" blood pressure as closely as phase V, is more difficult to detect, and has limited reproducibility. Many practitioners use phase V despite the guidelines. Universal adoption of phase V would improve the reliability of blood pressure measurements. PMID- 9734581 TI - Breast cancer and malpractice litigation. PMID- 9734580 TI - Proton pump inhibitors. PMID- 9734582 TI - Antenatal screening and prenatal diagnosis of thalassaemia: an update. PMID- 9734583 TI - Diseases associated with Helicobacter pylori. PMID- 9734584 TI - Positron emission tomography in assessing response to neoadjuvant chemotherapy for non-small-cell lung cancer. PMID- 9734585 TI - Primary pulmonary hypertension: new reasons for optimism? PMID- 9734586 TI - Yellow nails following brown snake bite. PMID- 9734587 TI - Severe tiger snake envenomation in a wilderness environment. PMID- 9734588 TI - Antivenoms and helicopter rescue services. PMID- 9734589 TI - Should third-generation cephalosporins be the empirical treatment of choice for severe community-acquired pneumonia in adults? PMID- 9734590 TI - Hypothermia induced by risperidone and olanzapine in a patient with Prader-Willi syndrome. PMID- 9734591 TI - Unemployment and young people's health. PMID- 9734592 TI - Future of medical training in Australia. PMID- 9734593 TI - Analgesic use and chronic renal failure: a critical review of the epidemiologic literature. AB - Heavy use of analgesics, particularly over-the-counter (OTC) products, has long been associated with chronic renal failure. Most of the earlier reports implicated phenacetin-containing analgesics as the risk factor. Since the early 1980s. several case-control studies have reported associations between chronic renal failure and use of other forms of analgesics, including acetaminophen, aspirin, and other non-steroidal antiinflammatory drugs (NSAIDs). Findings from these studies, however. should be interpreted with caution because of a number of inherent limitations and potential biases in the study design and data collection procedures. These limitations include: failure to identify patients early enough in the natural history of their disease to collect reliable information on analgesic use at an etiologically relevant time period; selection bias due to incomplete identification of subjects or low response rates; selection of cases and controls from different population bases; failure to employ survey techniques to improve reliability of recall of analgesic use; failure to collect detailed information on analgesic use such as year started and ended and reasons for switching analgesics; lack of standardization in the definition of regular analgesic use; and failure to adjust for phenacetin use and other confounding factors when assessing associations with analgesics other than those containing phenacetin. It is our hope that this review of study design limitations will lead to improvements in future studies of chronic renal failure risk. Since use of analgesics is widespread and new OTC products are introduced frequently, the potential impact of these drugs on the development of chronic renal failure may be significant, thus warranting continued evaluation of these products for any renal toxicity. PMID- 9734594 TI - Progression of glomerular diseases: is the podocyte the culprit? AB - The stereotyped development of the glomerular lesions in many animal models and human forms of progressive renal disease suggests that there are common mechanisms of disease progression. We propose the outline of such a mechanism based on following aspects: (1) The glomerulus is a complex structure, the stability of which depends on the cooperative function of the basement membrane, mesangial cells and podocytes, counteracting the distending forces originating from the high glomerular hydrostatic pressures. Failure of this system leads to quite uniform architectural lesions. (2) There is strong evidence that the podocyte is incapable of regenerative replication post-natally; when podocytes are lost for any reason they cannot be replaced by new cells. Loss of podocytes may therefore lead to areas of "bare" GBM. which represent potential starting points for irreversible glomerular injury. (3) Attachment of parietal epithelial cells to bare GBM invariably occurs when bare GBM coexists with architectural lesions, leading to the formation of a tuft adhesion to Bowman's capsule, the first "committed" lesion progressing to segmental sclerosis. (4) Within an adhesion the tuft merges with the interstitium, allowing filtration from perfused capillaries inside the adhesion towards the interstitium. The relevance of such filtration is as yet unclear but may play a considerable role in progression to global sclerosis and interstitial fibrosis. PMID- 9734595 TI - CLCN5 chloride-channel mutations in six new North American families with X-linked nephrolithiasis. AB - BACKGROUND: X-linked nephrolithiasis, or Dent's disease, encompasses several clinical syndromes of low molecular weight (LMW) proteinuria, hypercalciuria, nephrocalcinosis, nephrolithiasis, and renal failure, and is associated with mutations in the CLCN5 gene encoding a kidney-specific voltage-gated chloride channel. Some patients from Europe have rickets, and all symptomatic patients confirmed by mutation analysis have been male. METHODS: We analyzed the CLCN5 DNA sequence in six new families with this disease. RESULTS: In three probands, a single-base substitution yielded a nonsense triplet at codons 28, 34, and 343, respectively, and in two families, one of which was Hispanic, we found single base deletions at codons 40 and 44, leading to premature termination of translation. In the sixth family, a single-base change from C to T predicted substitution of leucine for serine at codon 244, previously reported in two European families with prominent rickets, though this patient of Ashkenazi origin did not have rickets. Each of these mutations was confirmed by restriction endonuclease analysis, or repeat sequencing and CFLP. The R34X mutation occurred in a Canadian infant with severe rickets. The family with the R28X nonsense mutation included one woman with recurrent kidney stones and another woman with glomerular sclerosis. In another family, a woman heterozygous for the W343X mutation also had nephrolithiasis. CONCLUSIONS: These studies expand the range of mutations identified in this disease, and broaden the phenotypic range to include clinically affected women and the first North American case with severe rickets. PMID- 9734596 TI - A model of autosomal recessive Alport syndrome in English cocker spaniel dogs. AB - BACKGROUND: Dogs with naturally occurring genetic disorders of basement membrane (type IV) collagen may serve as animal models of Alport syndrome. METHODS: An autosomal recessive form of progressive hereditary nephritis (HN) was studied in 10 affected, 3 obligate carrier, and 4 unaffected English cocker spaniel (ECS) dogs. Clinical, pathological, and ultrastructural features of the disease were characterized. Expression of basement membrane (BM) proteins was examined with an immunohistochemical technique using monospecific antibodies. RESULTS: Affected dogs had proteinuria and juvenile-onset chronic renal failure. Glomerular basement membrane (GBM) thickening and multilamellation typical of HN were observed in all renal specimens obtained from proteinuric dogs, and severity of GBM ultrastructural abnormalities varied with the clinical stage of disease. Expression of alpha3(IV) and alpha4(IV) chains was totally absent in the kidney of affected dogs. Expression of alpha5(IV) and a6(IV) chains was normal in Bowman's capsule, collecting tubular BM and epidermal BM of affected dogs. The alpha5(IV) chain was not expressed in distal tubular BM of affected dogs. Expression of alpha5(IV) chains was markedly reduced but not absent, and expression of alpha6(IV) chains was present in GBM of affected dogs. Expression of alpha1-alpha2(IV) chains in GBM of affected dogs was increased. Features of obligate carriers were similar to those of unaffected dogs. CONCLUSIONS: We conclude that HN in ECS dogs is a naturally occurring animal model of autosomal recessive Alport syndrome. However, it differs from human disease in the persistence of alpha5(IV) chains in GBM and in the appearance of a6(IV) chains in GBM. PMID- 9734597 TI - Novel mutations in the thiazide-sensitive NaCl cotransporter gene in patients with Gitelman syndrome with predominant localization to the C-terminal domain. AB - Gitelman syndrome (familial hypokalemia-hypomagnesemia syndrome) is an autosomal recessive inherited renal disorder characterized by defective tubular reabsorption of magnesium and potassium. In this study a group of 18 unrelated and 2 related Gitelman patients, collected from six different countries have been screened for mutations in the human thiazide-sensitive sodium-chloride cotransporter (SLC12A3) gene. Fourteen novel SLC12A3 mutations are presented along with six mutations described earlier, and three neutral polymorphisms. Among the tested patients are two who carry a total of three heterozygous SLC12A3 mutations. Two-thirds of the total number of mutant SLC12A3 alleles are amino acid substitutions. Most SLC12A3 gene mutations, 14 out of a total of 20, are localized at the intracellular carboxy-terminal domain of the NCCT protein. The pathogenicity of individual SLC12A3 mutations is based upon their predicted effect on SLC12A3 protein, and segregation in family members. Evolutionary conservation of substituted amino acid residues and their frequency in control chromosomes is presented. Identical mutations have been found in Gitelman families from different geographical origin, suggesting ancient mutations originating from a common ancestor. As yet, we have not found any evidence for a possible genotype-phenotype correlation. PMID- 9734598 TI - Expression of platelet-derived growth factor and its receptors in the developing and adult mouse kidney. AB - BACKGROUND: Experimental analysis of gene function is increasingly being accomplished using mouse models. Glomerular malformations occur in mice in which the platelet-derived growth factor (PDGF) B-chain gene or the PDGF receptor beta subunit gene have been deleted. To understand potential PDGF signaling pathways in the kidney, we determined the expression pattern of PDGF ligand and receptor genes in mouse kidney during development and in the mature adult kidney. METHODS: We used in situ hybridization to map the expression of transcripts encoding the PDGF ligands (A-chain and B-chain) and PDGF receptors (PDGFRalpha and PDGFRbeta) in the developing and mature kidney of the mouse. RESULTS: PDGF A-chain transcripts are expressed by epithelial cells (especially in what appear to be the loop of Henle) and possibly in vascular smooth muscle cells. Its receptor, PDGFRalpha, is expressed by interstitial cells. PDGF B-chain transcripts are most highly expressed by vascular endothelial cells of developing and adult kidney and minimally by visceral epithelia of immature glomeruli. PDGFRbeta transcripts are expressed by fetal blastemal cells, interstitial cells, mesangial cells, and vascular smooth muscle cells and by adult mesangial and interstitial cells. PDGFRalpha and PDGFRbeta expression is especially prominent in lipid-laden interstitial cells in the adult kidney. CONCLUSIONS: These patterns of expression are similar, but not identical, to those observed in rat and human and suggest that paracrine interactions mediated by the PDGF/PDGF receptor system may coordinate the development of the tubular, vascular, and interstitial components during kidney development and disease. PMID- 9734599 TI - Paracrine stimulation of human renal fibroblasts by proximal tubule cells. AB - Paracrine stimulation of human renal fibroblasts by proximal tubule cells. BACKGROUND: Interstitial fibrosis strongly predicts the degree and progression of renal failure in human renal disorders. Since active fibrosis tends to initially occur in a peritubular distribution, the possibility that human proximal tubule cells (PTC) relay fibrogenic signals to neighboring cortical fibroblasts was examined in vitro. METHODS: Cell proliferation (cell counts and thymidine incorporation), total collagen synthesis (proline incorporation), matrix metalloproteinase (MMP) activity (gelatin zymography), and autocrine secretion of insulin-like growth factor-I (IGF-I) were measured in primary cultures of human cortical fibroblasts cocultured with PTC or exposed to PTC-conditioned media (PTCCM). RESULTS: Cell numbers and thymidine incorporation rates were increased in cortical fibroblasts cocultured with PTC (136.4+/-7.3% and 119.3+/-8.2% of control values, respectively, P < 0.05) or incubated in PTC-CM (114.0+/-5.9%, P < 0.05 and 146.7+/-13.3%, P < 0.05, respectively). PTC-CM stimulated cortical fibroblast collagen synthesis (13.5+/-1.0% vs. 10.8+/-0.7%, respectively, N = 24, P < 0.05) and MMP-2 and MMP-9 secretion. Cortical fibroblast secretion of IGF-I binding protein-3 (IGFBP-3), which in turn modulates the autocrine and paracrine actions of IGF-I, was enhanced in the presence of PTC-CM compared with control (1162.2+/-94.2 vs. 969.1+/-58.9 ng/mg protein/day, P < 0.05), but no change was observed in cortical fibroblast secretion of IGFBP-2 (260.9+/-38.8 vs. 290.9+/ 36.6 ng/mg protein/day, P = NS) or IGF-I (56.7+/-6.6 vs. 57.0+/-6.8 ng/mg protein/day, P = NS). Human PTC secreted transforming growth factor-beta1 (TGF beta1) and the AB heterodimer of platelet-derived growth factor (PDGF-AB) in a time-dependent fashion and the augmentation of cortical fibroblasts mitogenesis, collagen synthesis and IGFBP-3 secretion induced by PTC-CM was replicated by exogenous TGF-beta1 and PDGF. Furthermore, the stimulatory effects of PTC on cortical fibroblasts were potentiated in transiently acidified PTC-CM (which activated latent TGF-beta1), and were abrogated by neutralizing antibodies specifically directed against TGF-beta1 and PDGF-AB. Cortical fibroblasts in turn released a soluble factor(s) into cortical fibroblast-conditioned media that reciprocally stimulated PDGF-AB production by PTC (4.79+/-1.55 vs. 0.78+/-.06 ng/mg protein/day, P < 0.05). CONCLUSIONS: PTC modulate the biological behavior of neighboring cortical fibroblasts in the human kidney through paracrine mechanisms, which include the production and release of PDGF-AB and TGF-beta1. Renal insults that result in proximal tubule injury may perturb this paracrine interaction, thereby culminating in excessive fibroblast proliferation and interstitial fibrosis. PMID- 9734600 TI - Effects of C-peptide on renal function at the early stage of experimental diabetes. AB - BACKGROUND: C-peptide has been reported to have biological effects on renal function early in the course of diabetes. This investigation was initiated to elucidate and amplify these findings with respect to protein leakage, hyperfiltration and renal functional reserve in diabetic rats. METHODS: Acute effects of 140 minutes i.v. infusion of human C-peptide (0.5 nmol x min(-1) x kg( 1) body wt) on renal function and urinary protein leakage were studied in anesthetized diabetic male rats two weeks after streptozotocin injection without insulin treatment. Streptozotocin-induced diabetic rats were studied with (N = 6) and without C-peptide (N = 11) treatment. The two groups showed a similarly elevated blood glucose concentration during the study. Age-matched normal rats served as controls (N = 5). Glomerular filtration rate (GFR) was measured by inulin clearance in the basal state and during a 60-minute glycine infusion (0.22 mmol x min(-1) x kg(-1) body wt)-resembling a protein load challenge-to test the renal functional reserve in all three groups. RESULTS: In the basal state, the non-C-peptide-treated diabetic rats displayed increased GFR and increased total protein leakage compared with normal rats. Whereas normal rats responded to glycine infusion with an increase in GFR, no increase occurred in diabetic rats not treated with C-peptide. In diabetic rats given C-peptide, this reduced the initial glomerular hyperfiltration prior to glycine infusion. This indicates a specific effect, since a control peptide with the same amino acid composition as C-peptide, but in a randomized sequence, had no such effect. C-peptide also restored half of the normal renal functional reserve and resulted in 70% lower (P < 0.05) total protein leakage compared with that in rats not given C-peptide. CONCLUSION: Thus, short-term infusion of C-peptide had beneficial effects on protein leakage and hyperfiltration and improved the renal functional reserve in rats with experimental diabetes. PMID- 9734601 TI - Alpha-melanocyte-stimulating hormone inhibits renal injury in the absence of neutrophils. AB - BACKGROUND: We previously showed that alpha-melanocyte stimulating hormone (alpha MSH) decreases ischemia/reperfusion injury even when started six hours after ischemia. Alpha-MSH inhibits both neutrophil accumulation and nitric oxide production. To determine the relative importance of alpha-MSH on the neutrophil pathway, we examined the effects of alpha-MSH in injury models where neutrophil effects are minimal or absent. METHODS: We studied the effects of alpha-MSH in (1) intercellular adhesion molecule-1 (ICAM-1) knock-out and background mice that were subjected to 40 minutes of ischemia and 24 hours reperfusion, and (2) isolated kidneys that were subjected to in vivo ischemia for 20 minutes and then perfused ex vivo for one hour without neutrophils. To begin to search for direct tubule effects of alpha-MSH, we studied the effect of alpha-MSH on nitric oxide (NO) in endotoxin/interferon-gamma-treated mouse cortical tubule cells. RESULTS: ICAM-1 knock-out mice had 75% less neutrophil infiltration than background mice after ischemia. Despite the relative lack of neutrophils, alpha-MSH inhibited renal injury in ICAM-1 knock-out mice. Alpha-MSH also significantly preserved GFR and tubular sodium reabsorption in the isolated perfused ischemic kidney model. Alpha-MSH and a nitric oxide inhibitor did not exhibit synergy. Finally, alpha MSH inhibited nitrite production by 20% in the mouse cortical tubule cells (MCT), similar to parallel observations in a cultured mouse macrophage line (RAW cells). CONCLUSIONS: We conclude that alpha-MSH decreases renal injury when neutrophil effects are minimal or absent, indicating that alpha-MSH inhibits neutrophil independent pathways of renal injury. The preservation of sodium absorption ex vivo and inhibition of nitrite production in cultured MCT cells suggests that alpha-MSH inhibits tubular injury by direct tubular effects. PMID- 9734602 TI - Angiotensin II induces superoxide anion production by mesangial cells. AB - BACKGROUND: The recognized role of angiotensin II (Ang II) in the pathogenesis of the progression of renal disease cannot be solely attributed to Ang II's hemodynamic effects. Indeed, growth stimulating signals driven by Ang II promote mesangial cell (MC) hypertrophy and extracellular matrix production, prominent features of progressive glomerular injury. Superoxide anion (O2-) avidly interacts with nitric oxide, an endogenous vasodilator that inhibits growth factor stimulated MC growth and matrix production. In addition, O2- acting as an intracellular signal is linked to growth related responses such as activation of mitogen activated protein (MAP) kinases. The studies reported herein were designed to investigate: (a) whether Ang II induces MC O2-production and (b) if increased O2- production elicits growth responses in MC. METHODS: MC were exposed to Ang II for 24 or 48 hours. In some experiments, in addition to Ang II, MC were exposed to: diphenylenieodonium (DPI), an inhibitor of the flavin containing NADH/NADPH oxidase; losartan (LOS), an Ang II type 1 (AT1) receptor blocker; PD 98059, a MAP kinases inhibitor; the protein kinase C inhibitors Calphostin C or H 7; and the tyrosine kinase inhibitors, herbymycin A or genistein. RESULTS: Ang II (10(-5) M to 10(-8) M) dose dependently increased MC O2- production up to 125% above control (ED 50 5 x 10(-7) M). LOS as well as DPI, and the PKC inhibitors blocked Ang II stimulated MC O2- production. Ang II dose dependently increased MC 3H-leucine incorporation, and MC protein content, two markers of MC hypertrophy, as well as 3H-thymidine incorporation, a marker of MC hyperplasia. PD98059, a specific inhibitor of MAP kinases prevented Ang II induced MC hypertrophy. Moreover, LOS, DPI, and the PKC inhibitors each independently inhibited MC 3H leucine incorporation, thereby establishing the specificity of Ang II induced O2- in driving MC hypertrophy. CONCLUSIONS: The current studies demonstrate a previously unrecognized link between Ang II and MC O2- production that may participate in the pathophysiology of progressive renal disease by concomitantly affecting the hemodynamics of the glomerular microcirculation as well as growth related responses of MC to injury. PMID- 9734603 TI - Molecular mechanism(s) of action of norepinephrine on the expression of the angiotensinogen gene in opossum kidney cells. AB - BACKGROUND: Norepinephrine (NE) is the major endogenous neurotransmitter of the renal sympathetic nerves interacting with both the alpha- and beta-adrenoceptors in the renal proximal tubules. We have previously reported that isoproterenol and iodoclonidine stimulate the expression of the angiotensinogen (ANG) gene in opossum kidney (OK) proximal tubular cells via the beta1-adrenoceptor and alpha2 adrenoceptor, respectively. We hypothesized that NE may interact with the beta- and/or alpha2-adrenoceptors to stimulate the expression of the ANG gene in OK cells. METHODS: The fusion genes containing the various lengths of the 5' flanking regulatory sequence of the rat ANG gene fused with a human growth hormone (hGH) gene as a reporter were stably transfected into the OK cells. The stimulatory effect of NE on the expression of the fusion genes was evaluated by the amount of immunoreactive hGH (IR-hGH) secreted into the culture medium. RESULTS: The addition of NE stimulated the expression of the fusion gene, pOGH (ANG N-1498/+18) in a dose-dependent manner. The stimulatory effect of NE was inhibited in the presence of propranolol, atenolol, Rp-cAMP, yohimbine, staurosporine, H-7 and U73122 but not in the presence of ICI 118,551 and prazosin. The addition of a combination of isoproterenol and iodoclonidine synergistically stimulated the expression of pOGH (ANG N-1498/+18) as compared to the addition of isoproterenol and iodoclonidine alone. Furthermore, the addition of NE, forskolin, 8-Br-cAMP or phorbol 12-myristate (PMA) stimulated the expression of pOGH (ANG N-806/-779/-53/+18), a fusion gene containing the putative cAMP responsive element (CRE, ANG N-806/-779) upstream of the ANG promoter (ANG N-53/+ 18) in OK 95 cells, but had no effect on the expression of fusion genes containing the mutant of the CRE. CONCLUSION: These studies demonstrate that the stimulatory effect of NE on the expression of the ANG gene in OK cells may be mediated via both the beta1- and alpha2-adrenoceptors and via the CRE (ANG N-806/-779) in the 5'flanking region of rat ANG gene. PMID- 9734605 TI - Inhibition of the matrix metalloproteinase system in a rat model of chronic cyclosporine nephropathy. AB - BACKGROUND: Chronic cyclosporine A (CsA)-induced nephropathy is histologically characterized by tubular lesions, the interstitial recruitment of inflammatory cells, arteriolopathy and focal interstitial fibrosis. Recent studies show that the intrarenal inhibition of matrix degradation and recruitment of monocytes/ macrophages into the kidney plays a critical role in the development of renal interstitial fibrosis. METHODS: We examined the expression of components of the matrix metalloproteinase (MMP) system and plasminogen activator inhibitor type-1 (PAI-1) in kidneys from rats injected daily s.c. during three weeks with CsA (10, 15 or 20 mg CsA/kg body wt) or vehicle solution. RESULTS: In all CsA-treated rats, serum creatinine levels were significantly elevated compared to control levels. The extent of CsA-induced atrophy was not influenced by the dosage during a three-week CsA treatment. The administration of CsA did not significantly increase total cortical interstitial collagen deposition, whereas alpha-smooth muscle actin expression was significantly increased in all CsA-treated rats. Analysis of the different subpopulations of inflammatory cells recruited into the chronically injured kidney revealed a marked influx of macrophages into fibrotic cortical foci of CsA-treated rats. The number of cortical macrophages was highest in the group receiving the highest CsA dose. PAI-1 antigen, present in proximal tubular lysosomes in kidneys from all experimental groups, stained very intensely in atrophic tubules in CsA-treated rats. Both stromelysin and interstitial collagenase mRNA were expressed in the kidneys of control rats, but their message transcription remained unaltered after CsA treatment. In contrast, the expression of tissue inhibitor of matrix metalloproteinase type 1 (TIMP-1) was significantly increased after CsA treatment. TIMP-1 mRNA was undetectable in renal sections from sodium-depleted vehicle-treated animals using the in situ hybridization (ISH) technique. ISH of selected renal sections of CsA-treated rats identified the cells responsible for the increased TIMP-1 message transcription after CsA administration, mainly as interstitial cells and also as visceral and parietal epithelial cells. CONCLUSIONS: These results suggest that the locally increased expression of TIMP-1 rather than a decrease of matrix metalloprotease expression, contributes to the development of CsA-induced focal interstitial fibrosis in the rat. PMID- 9734604 TI - Direct nucleation of calcium oxalate dihydrate crystals onto the surface of living renal epithelial cells in culture. AB - BACKGROUND: The interaction of the most common crystal in human urine, calcium oxalate dihydrate (COD), with the surface of monkey renal epithelial cells (BSC-1 line) was studied to identify initiating events in kidney stone formation. METHODS: To determine if COD crystals could nucleate directly onto the apical cell surface, a novel technique utilizing vapor diffusion of oxalic acid was employed. Cells were grown to confluence in the inner four wells of 24-well plates. At the start of each experiment, diethyloxalate in water was placed into eight adjacent wells, and the plates were sealed tightly with tape so that oxalic acid vapor diffused into a calcium-containing buffer overlying the cells. RESULTS: Small crystals were visualized on the cell surface after two hours, and by six hours the unambiguous habitus of COD was confirmed. Nucleation onto cells occurred almost exclusively via the (001) face, one that is only rarely observed when COD crystals nucleate onto inanimate surfaces. Similar results were obtained when canine renal epithelial cells (MDCK line) were used as a substrate for nucleation. Initially, COD crystals were internalized almost as quickly as they formed on the apical cell surface. CONCLUSIONS: Face-specific COD crystal nucleation onto the apical surface of living renal epithelial cells followed by internalization is a heretofore unrecognized physiological event, suggesting a new mechanism to explain crystal retention within the nephron, and perhaps kidney stone formation when this process is dysregulated or overwhelmed. PMID- 9734606 TI - Immunohistochemical and serological evidence for the role of streptococcal proteinase in acute post-streptococcal glomerulonephritis. AB - BACKGROUND: We have previously demonstrated the preferential secretion of streptococcal proteinase or streptococcal pyrogenic exotoxin B (SPEB) by nephritic strains of Group A streptococci isolated from the skin or throat of patients with acute poststreptococcal glomerulonephritis (APSGN). METHODS: To further explore the possible role of SPEB in APSGN, we performed ELISA studies to detect anti-SPEB antibodies in the sera of patients with APSGN, acute rheumatic fever (ARF), scarlet fever (SF) and normal children. Using ELISA, anti-SPEB titers on acute and convalescent APSGN sera were measured to determine immunity to APSGN. We also performed immunofluorescence studies on APSGN and non-APSGN kidney biopsies to probe for the presence and localization of SPEB. RESULTS: Our data show that anti-SPEB antibodies are present in APSGN sera and antibody titers are significantly higher than in ARF, SF and normal sera. Anti-SPEB titers tend to rise acutely and decrease with time but do not reach baseline after one year. When kidney biopsies were probed with rabbit anti-SPEB antibody, 12 of 18 (67%) of the APSGN cases were positive while only 4 of 25 (16%) of the non-APSGN cases were positive. CONCLUSIONS: In summary, we were able to demonstrate unique reactivity to SPEB in human sera and kidney biopsies of APSGN suggesting a significant role of this toxin in the pathogenesis of acute post-streptococcal glomerulonephritis. PMID- 9734607 TI - In situ analysis of C-C chemokine mRNA in human glomerulonephritis. AB - BACKGROUND: Glomerular and tubulointerstitial accumulations of macrophages and T cells are a prominent feature of immune inflammatory glomerulonephritis. The C-C family of chemokines are major mononuclear-cell chemoattractants and may be central to the recruitment of these cells. METHODS: Using in situ hybridization (ISH) we analyzed the expression of mRNA for the C-C chemokines monocyte chemoattractant protein-1 (MCP-1), macrophage inflammatory protein-1alpha and beta (MIP-1alpha, MIP-1beta) and RANTES in renal biopsy material from twenty patients with glomerulonephritis. RESULTS: In overt inflammatory glomerulonephritides, chemokine transcripts were differentially expressed by glomerular and tubulointerstitial leukocyte infiltrates, glomerular parietal and proximal tubular epithelial cells and endothelial cells. There was little expression in minimal change nephropathy and normal tissue. Expression of individual chemokines correlated with intrarenal T cell and macrophage infiltrates. Combined immunohistochemistry and ISH demonstrated that 56.9% of cells expressing MCP-1 mRNA were CD68+ve (monocytes/macrophages) and 53% of infiltrating CD68 +ve cells were MCP-1 mRNA positive. CONCLUSIONS: These studies indicate that the in situ production of C-C chemokines by resident and infiltrating cells may play a crucial role in regulating macrophage and T-cell recruitment in glomerulonephritis. PMID- 9734608 TI - IgA induced activation of human mesangial cells: independent of FcalphaR1 (CD 89). AB - BACKGROUND: IgA nephropathy (IgAN) is characterized by deposition of polymers of IgA1 in the mesangium, accumulation of mesangial matrix and mesangial cell proliferation. Activation of the mesangial cell by IgA, via an IgA receptor, may be an initiating event in the pathology of IgAN. METHODS: We examined the ability of radiolabeled, normal serum IgA1 to bind human mesangial cells (HMC). Activation of HMC by monomeric (mIgA1) and heat aggregated IgA1 (AIgA1) was compared by Northern analysis of c-jun expression. The expression of FcalphaR1 (CD89) mRNA on our cultured mesangial cells was also assessed by Northern analysis, reverse transcription-polymerase chain reaction (RT-PCR) and flow cytometry. RESULTS: 125I-mIgA1 and 125I-AIgA1 bound to HMC in a dose-dependent, saturable manner with similar affinities. There were 1.2 x 10(6) binding sites per cell, with an affinity constant of 2.3 x 10(6) M(-1). AIgA1 induced c-jun expression in a time and dose-dependent manner (2.4-fold above baseline after 60 min exposure to AIgA1 200 microg/ml) while mIgA1 had no effect on c-jun expression. No message for CD 89 was detectable in quiescent or AIgA1 stimulated HMC by Northern analysis or RT-PCR using several primer sequences based on the sequence of U937 FcalphaR cDNA. Flow cytometry on the mesangial cells, using My 43, a monoclonal antibody to FcalphaR1 confirmed that CD 89 was not present on the cell. CONCLUSION: These results demonstrate that HMC bind mIgA1 and AIgA1 with similar affinity. However, activation of HMC requires an aggregated form of IgA1. These processes are independent of FcalphaR1, suggesting the presence of a new IgA receptor on mesangial cells. PMID- 9734609 TI - Identification of oxidized low density lipoprotein in human renal biopsies. AB - BACKGROUND: Intraglomerular lipid deposition is frequently observed in routine renal biopsies, and it has been suggested that lipid peroxidation of low density lipoprotein (LDL) may be implicated in the pathogenesis of progressive glomerulosclerosis. We have examined whether oxidized LDL (Ox-LDL) is present in the glomeruli of patients with renal disease and whether intrinsic human glomerular cells express NADPH-oxidase (a superoxide-generating enzyme found in professional phagocytes). METHODS: Immunocytochemical study was performed on 939 renal biopsy specimens, using monoclonal antibodies (mAbs) OL-10, 48 and 449, and polyclonal antibody against human apolipoprotein (apo) B. Mouse mAb OL-10 recognizes malondialdehyde (MDA)-modified peptide epitope, and mAbs 48 and 449 react with alpha and beta subunits of cytochrome b558, an essential component of NADPH-oxidase. RESULTS: Sixty-two (6.6%) of the 939 patients with renal disease exhibited a staining for MDA-altered protein or Ox-LDL in the glomeruli, mainly in the sclerotic segments or mesangial areas. Group 1 patients with heavy Ox-LDL deposition mainly in the sclerotic segments showed a higher frequency of renal insufficiency and heavy proteinuria and a greater degree of glomerulosclerosis, compared to those in group 2 with mesangial Ox-LDL staining. The distribution of MDA protein epitopes, in general, paralleled the deposition of apo B epitopes. Immunoelectron microscopy of ultrathin frozen sections showed the presence of immunogold particles for mAbs 48 and 449 in the cytoplasm of resident glomerular cells of both normal and diseased kidneys. When immunoblotted with mAb OL-10, one band from the IgA nephropathy and focal segmental glomerulosclerosis groups at approximately 260 kD was labeled, whereas immunostaining of normal control samples revealed no staining. CONCLUSIONS: These results indicate that Ox-LDL is present mainly in the lesions of glomerulosclerosis and mesangial areas in human renal biopsies. They also suggest that patients with heavy Ox-LDL accumulation in the sclerotic segments of glomeruli have more advanced renal disease than those with mesangial Ox-LDL and that resident glomerular cells generate cytochrome b558, the potential of which may not suffice to induce peroxidation of LDL in the diseased glomeruli. PMID- 9734610 TI - Protection from renal ischemia-reperfusion injury by the 2-methylaminochroman U83836E. AB - BACKGROUND: In a prior study the 21-aminosteroid (lazaroid) U74389F provided in vivo protection from oxidative stress when used as a preventive therapy in ischemia-reperfusion injury in the kidney. As the cell membrane is the principal site for lipoperoxidation, in the current study the very lipophilic 2 methylaminochroman U83836E, a recently developed lazaroid, was administered to rats at 3 mg/kg before renal ischemia-reperfusion. In addition to the biochemical parameters, the renal function and the histological appearance were carefully evaluated. METHODS: Glutathione, adenine nucleotides and lipid peroxidation products were determined in kidneys reperfused for 2 and 24 hours after 90 minutes of ischemia. Renal function was assessed by plasma creatinine, and renal injury by histological examination. RESULTS: Reperfusion-induced glutathione oxidation, expressed as an oxidized-to-total glutathione ratio, was significantly attenuated both after 2 and 24 hours of reperfusion by treatment with U83836E. Adenosine triphosphate (ATP) was still significantly depleted after 24 hours in the control group, while at the same time treated animals had already recovered to baseline values. Lipid peroxidation products were significantly lower in lazaroid-groups both after 2 and 24 hours of reperfusion. Renal function after 24 hours of reperfusion was notably better in the treated rats. Histological examination confirmed the protective action of the drug. After 24 hours the control group showed large areas of parenchymal hemorrhage and necrosis with dilated tubules and blood vessel thrombosis, while treated animals showed small necrotic areas with a background of mild interstitial inflammatory cells. CONCLUSIONS: Our results suggest that there is a protective effect of U83836E in ischemia-reperfusion injury, in that tissue damage due to oxidative stress is reduced, thus ameliorating renal function impairment. PMID- 9734611 TI - Tubular epithelial-myofibroblast transdifferentiation in progressive tubulointerstitial fibrosis in 5/6 nephrectomized rats. AB - BACKGROUND: Tubulointerstitial fibrosis is the final common pathway to end-stage renal failure. The present study investigated the potential role of tubular epithelial cells (TEC) in progressive fibrosis in the rat remnant kidney model. METHODS: Rats underwent 5/6 nephrectomy or a sham operation (control), and groups of six animals were killed at weeks 1, 3, 5, 9, 13, 17 and 21. RESULTS: Immunohistochemistry staining and in situ hybridization at week 3 after nephrectomy demonstrated de novo expression of alpha-smooth muscle actin (alpha SMA)--a marker of smooth muscle cells and myofibroblasts--by TEC that was invariably associated with disruption of the tubular basement membrane (TBM). This phenotypic evidence of tubular epithelial-myofibroblast transdifferentiation was supported by ultrastructural studies identifying the presence of characteristic actin microfilaments and dense bodies within TEC with a transformed morphology. In the late stage of this apparent tubular epithelial myofibroblast transdifferentiation, TEC lost apical-basal polarity and tight junctions, became elongated, detached from the TBM, separated from neighboring cells and appeared to migrate into the peritubular interstitium through the damaged basement membrane. Indeed, focal peritubular accumulation of alpha-SMA+ myofibroblasts and local tubulointerstitial fibrosis was closely associated with alpha-SMA+ tubules, suggesting a tubular epithelial origin for some of these cells. Quantitative analysis found a significant correlation between the number of alpha-SMA+ TEC and the accumulation of interstitial alpha-SMA+ myofibroblasts and the severity of tubulointerstitial fibrosis (both P < 0.001). CONCLUSIONS: This study provides phenotypic and morphological evidence to support the hypothesis that TEC are pro-fibrogenitor cells capable of tubular epithelial myofibroblast transdifferentiation in progressive renal fibrosis. In addition, we postulate that disruption of the TBM, which facilitates epithelial cell contact with the interstitial matrix, promotes this process of transdifferentiation. PMID- 9734612 TI - Differential upregulation of rat Na-K-Cl cotransporter, rBSC1, mRNA in the thick ascending limb of Henle in different pathological conditions. AB - BACKGROUND: Na-Cl cotransport across the apical membrane of epithelial cells in the thick ascending limb of the loop of Henle (TAL) plays a major role in salt accumulation for hypertonic medullary interstitium. The electroneutral, rat bumetanide-sensitive sodium transporter, rBSC1, is involved in this process. We studied the level of rBSC1 mRNA in dehydration and cardiac failure, since sodium transport in TAL may be enhanced in both conditions in spite of the difference in extracellular fluid accumulation. METHODS: Male Sprague-Dawley rats were deprived of water for 24 hours and myocardial infarction of about 40% of left ventricular circumference was induced in another group of rats that later developed congestive heart failure (CHF). Digoxigenin-labeled cRNA probe for rBSC1 was constructed using polymerase chain reaction (PCR), and Northern blot analysis was performed using RNAs from renal outer medulla. By inducing a point mutation at the middle of PCR product, we compared the amount of rBSC1 transcripts in the renal cortex using competitive PCR, since TAL represents a small fraction of the total cortical tissue. RESULTS: Northern analysis showed a significant increase in rBSC1 mRNA in the renal outer medulla of both dehydrated and CHF rats. In the renal cortex, however, the increase was noted only in CHF by competitive PCR. In situ hybridization using the riboprobe for northern analysis demonstrated that the transcript signal in dehydrated rats was intensified segmentally in TAL located in the inner stripe of outer medulla. Western analysis and immunohistochemistry using a specific antibody against rBSC1 confirmed the distinct segmental enhancement of apical protein expression in dehydration and diffuse enhanced expression in CHF. CONCLUSIONS: rBSC1 is differentially upregulated in different pathological conditions. PMID- 9734613 TI - Differential effects of calcium channel blockers on size selectivity of proteinuria in diabetic glomerulopathy. AB - BACKGROUND: Calcium channel blockers (CCBs) are known to have differential effects on both changes in proteinuria as well as progression of diabetic nephropathy. No clinical study, however, has evaluated whether the differential antiproteinuric effects of CCBs may be explained by their effect on glomerular membrane permeability. We, therefore, tested the hypothesis that certain subclasses of CCBs reduce proteinuria by changing size selectivity of the glomerular membrane, hence changing its permeability. METHODS: Twenty-one patients with type 2 diabetes and the presence of nephropathy with hypertension were randomized to receive either diltiazem CD or nifedipine GITS after baseline data for mean systolic and diastolic pressure, urinary protein excretion, glomerular filtration rate, renal plasma flow, neutral dextran and IgG clearances were obtained. Glomerular filtration rate, renal plasma flow, neutral dextran and IgG clearance were measured every three months, arterial pressure and heart rate every month. Patients were followed for 21 months. RESULTS: At 21 months, both patient groups had similar levels of blood pressure control, however, only the diltiazem group had a change in proteinuria (4+/-10%delta, nifedipine vs. -57+/ 18%delta, diltiazem; P < 0.001) with improvement in glomerular size selectivity and change in IgG clearance. CONCLUSIONS: These data support the hypothesis that CCBs that provide sustained reductions in proteinuria do so, in part, by improving glomerular size permselectivity. PMID- 9734614 TI - NCX1 Na/Ca exchanger inhibition by antisense oligonucleotides in mouse distal convoluted tubule cells. AB - BACKGROUND: Plasma membrane NCX1 Na+/Ca2+ exchangers mediate cellular Ca2+ efflux. Renal distal convoluted tubule (DCT) cells express transcripts encoding three alternatively spliced NCX1 isoforms: NACA2 (exons B, C, D), NACA3 (exons B and D), and NACA6 (exons A, C, D). We used antisense oligodeoxynucleotides (ODNs) to determine the function of these NACA isoforms on Na+/Ca2+ exchanger activity and expression in DCT cells. METHODS: Sense and antisense ODNs targeting exchanger transcripts were introduced into DCT cells permeabilized with streptolysin O. Na+/Ca2+ exchange activity was assessed by measuring Na+ dependent changes of free intracellular Ca2+ concentration (delta[Ca2+]i), in single cells, when the electrochemical gradient for Na+ was reversed. RESULTS: The change of [Ca2+]i in cells treated with antisense ODNs to a downstream or upstream region common to all NCX1 isoforms was 173 nM (-66%) to the downstream region located in the putative ninth transmembrane domain, and 226 nM (-39%) with ODNs to an upstream region located 5' to the variable portion of the intracellular loop. Antisense ODNs to exon B, present in both NACA2 and NACA3, decreased delta[Ca2+]i by 209 nM (-44%), while antisense ODNs specific for NACA6 (exon A) were without effect. Antisense ODNs specific for exon C, present in NACA2 and NACA6, decreased delta[Ca2+]i by 226 nM (-39%). Northern analysis of mRNA prepared from primary cultures of distal tubule cells revealed exon B- but not exon A-containing transcripts. Immunofluorescence analysis using a polyclonal antibody that recognizes NCX1 confirmed that protein expression was inhibited after treatment with the exon B antisense ODNs. CONCLUSION: These findings show that Na+-dependent cellular Ca2+ efflux in DCT cells is primarily mediated by NACA2 and NACA3. PMID- 9734616 TI - Association of thin basement membrane nephropathy with hypercalciuria, hyperuricosuria and nephrolithiasis. AB - BACKGROUND: Familial persistent microhematuria with normal renal function is the most common presentation of thin basement membrane nephropathy (TBMN). Gross hematuria episodes and loin pain attacks are other manifestations of the disease. On the other hand, it has been shown that hypercalciuria (HC) and hyperuricosuria (HU) can produce both gross or microscopic non-glomerular hematuria, in addition to their role in renal stone formation. METHODS: We studied the prevalence of HC, HU and nephrolithiasis in a group of 27 biopsy-proven TBMN as well as in 19 non biopsied first-degree relatives with persistent microhematuria and 25 first degree relatives without microhematuria. A group of 27 patients with IgA nephropathy (IgAN) and persistent microhematuria, and another group of 20 healthy subjects without known renal diseases were selected as control groups. RESULTS: Ten (37%) patients with TBMN and 8 (42%) relatives with microhematuria showed HC and/or HU at presentation; relatives without microhematuria, IgAN patients and normal controls showed a significantly lower prevalence of HC and HU. The prevalence of previous nephrolithiasis among TBMN patients (25%) was significantly higher than in IgAN patients (3%; P < 0.05). Family history of nephrolithiasis was recorded in 14 (51%) of the 27 TBMN families, in contrast with 2 of 27 (7%) with IgAN and 1 of 20 (5%) in normal controls (P < 0.05). The prevalence of nephrolithiasis, gross hematuria bouts and loin pain episodes among TBMN patients and microhematuric relatives showing HC and/or HU at presentation (44%, 44% and 27%, respectively) were significantly higher than those of TBMN patients and microhematuric relatives with normal calcium and uric acid urinary excretions (10%, 7% and 3%, respectively; P < 0.05). At the end of follow-up (8.8+/-4.1 years in TBMN patients and 9.1+/-4.2 years in relatives with microhematuria), all the cases maintained normal renal function. CONCLUSIONS: We found a high prevalence of HC, HU, and nephrolithiasis among TBMN patients and relatives with microhematuria. Our study also shows a significant relationship between the presence of HC and/or HU and the prevalence of nephrolithiasis, gross hematuria bouts and loin pain episodes. PMID- 9734615 TI - Intermittent calcitriol therapy in secondary hyperparathyroidism: a comparison between oral and intraperitoneal administration. AB - BACKGROUND: Intermittent oral or intravenous doses of calcitriol given two or three times per week are commonly used to treat secondary hyperparathyroidism (secondary HPT). This study was undertaken to compare the biochemical and skeletal responses to thrice weekly intraperitoneal (i.p.) versus oral doses of calcitriol in children with secondary HPT undergoing peritoneal dialysis (CCPD). METHODS: Forty-six patients aged 12.5+/-4.8 years on CCPD for 22+/-25 months were randomly assigned to treatment with oral (p.o.) or i.p. calcitriol for 12 months; 17 subjects given p.o. calcitriol and 16 subjects given i.p. calcitriol completed the study. Bone biopsies were performed at the beginning and at the end of the study, while determinations of serum and total ionized calcium, phosphorus, alkaline phosphatase, parathyroid hormone (PTH) and calcitriol levels were done monthly. RESULTS: Serum total and ionized calcium levels were higher in subjects treated with i.p. calcitriol, P < 0.0001, whereas serum phosphorus levels were higher in those given p.o. calcitriol, P < 0.0001. For the i.p. group, serum PTH levels decreased from pre-treatment values of 648+/-125 pg/ml to a nadir of 169+/ 57 pg/ml after nine months. In contrast, serum PTH levels did not change from baseline values of 670+/-97 pg/ml in subjects given p.o. calcitriol, P < 0.0001 by multiple regression analysis. Serum alkaline phosphatase levels were also lower in patients treated with i.p. calcitriol, P < 0.0001, but there was no difference between groups in the average dose of calcitriol given thrice weekly. The skeletal lesions of secondary HPT improved in both groups, 33% of patients developed adynamic bone lesion. CONCLUSION: Differences in the bioavailability of calcitriol and/or in phosphorus metabolism may account for the divergent biochemical response to p.o. and i.p. calcitriol. PMID- 9734617 TI - Plasma calcium-oxalate saturation in children with renal insufficiency and in children with primary hyperoxaluria. AB - BACKGROUND: Calcium-oxalate (CaOx) deposition and systemic oxalosis are uncommon in children with chronic renal failure (CRI), but frequent in children with primary hyperoxaluria type I (PH-1). We hypothesized a difference in plasma CaOx saturation (betaCaOx) and its determining factors would explain this discrepancy. METHODS: Therefore, in addition to common biochemical measurements, plasma oxalate (POx), citrate (PCit) and sulfate (PSulf) (plasma anions) were measured and betaCaOx was calculated in 17 PH-1 patients with normal renal function receiving pyridoxine and citrate therapy, in 54 children with CRI (SCr 0.9 to 5.9 mg/dl), and in 50 healthy children (NL). Plasma anions were analyzed by ion chromatography and betaCaOx was calculated using a PC-based program for solution equilibria. RESULTS: Compared to NL, all plasma anion levels and betaCaOx were higher in PH-1 and CRI; POx, PCit and betaCaOx were higher in PH-1 than in CRI (P < 0.05), but PSulf was higher in CRI (P < 0.01). BetaCaOx and POx were correlated in all groups (r = 0.63 to 0.95, P < 10(-4)). POx and betaCaOx were both inversely correlated to a decrease in GFR in CRI patients. PCit and PSulf did not influence betaCaOx. Although supersaturation (betaCaOx > 1) was found in 7 CRI and in 4 PH-1 patients, eye examinations were suspicious for CaOx depositions only in the PH-1 patients, while systemic oxalosis was confirmed in one PH patient because of oxalate osteopathy. CONCLUSIONS: In PH-1, POx and betaCaOx are elevated even with normal renal function, which increases the likelihood of CaOx crystal deposition. Therefore, more effective therapy to decrease betaCaOx is crucial to reduce the risk of systemic oxalosis. In children with CRI unknown, but presumably protective substances, help prevent the risk of systemic oxalosis, despite increased POx and betaCaOx levels, often to supersaturation levels. PMID- 9734618 TI - Smoking as a risk factor for end-stage renal failure in men with primary renal disease. AB - BACKGROUND: It is not known whether smoking increases the risk of end-stage renal failure (ESRF) in patients with primary renal disease. METHODS: We performed a retrospective multicenter case-control study including 582 patients from nine centers in Germany, Italy and Austria. The diseases investigated were IgA glomerulonephritis (IgA-GN) as a model of inflammatory renal disease and autosomal dominant polycystic kidney disease (ADPKD) as a model of non inflammatory renal disease. Cases were patients who had progressed to ESRF and controls were patients who were not in ESRF, that is, whose serum-creatinine failed to progress to >3 mg/dl during the observation period and who did not require renal replacement therapy. Matching for renal disease (IgA-GN, ADPKD), gender, age at renal death and region of residence resulted in 102 individually matched pairs (IgA-GN N = 54, ADPKD N = 48). Multiple conditional logistic regression was used to estimate adjusted odds ratios for independent tobacco effects. RESULTS: In men (matched pairs: IgA-GN N = 44, ADPKD N = 28), a significant dose-dependent increase of the risk to progress to ESRF was found (non-adjusted). The baseline risk was defined as <5 pack-years (PY): (i) 5 to 15 PY, odds ratio 3.5 (95% CI 1.3 to 9.6), P = 0.017; (ii) >15 PY = 5.8 (2.0 to 17), P = 0.001. Systolic blood pressure, ACE inhibitor treatment and age at diagnosis emerged as potential confounders. After adjustment, the risk for ESRF in men with >5 PY was highly increased for patients without ACE inhibitor treatment [10.1 (2.3 to 45), P = 0.002] but not with ACE inhibitor treatment [1.4 (0.3 to 7.1), P = 0.65]. CONCLUSION: Smoking increases the risk of ESRF in men with inflammatory and non-inflammatory renal disease. PMID- 9734619 TI - Acute leptin regulation in end-stage renal failure: the role of growth hormone and IGF-1. AB - BACKGROUND: Leptin, a recently discovered peptide involved in nutrient intake and energy expenditure, has been shown to be abnormally regulated in certain conditions such as obesity. In chronic renal failure, leptin appears to be increased. However, little is known about leptin regulation during chronic renal failure (CRF). METHODS: We measured serum leptin in eight well nourished, chronic hemodialysis patients (seven males, one female) receiving anabolic factors for three days as either recombinant insulin-like growth factor-1 (rhIGF-1) or a combination of recombinant growth hormone (rhGH) plus recombinant IGF-1, in a random cross-over trial. RESULTS: Serum leptin values were in the range of normal volunteers matched for body mass index. As reported in other conditions, serum leptin was strongly correlated with patients dry body wt (P = 0.01) and body fat (P = 0.0001). Both treatments affected serum leptin in a rapid and opposite manner. RhIGF-1 decreased serum leptin from 11.2+/-20.8 (SD) to 4.3+/-3.8 microg/liter (P = 0.011), whereas the combination of rhGH + rhIGF-1 increased serum leptin from 7.4+/-9.4 to 21.0+/-32.9 microg/liter (P = 0.011). Regression analyses indicated a linear regression between serum leptin and insulin variations after treatment. CONCLUSIONS: This study shows for the first time that both rhIGF-1 and rhGH acutely regulate serum leptin in dialysis patients. Whether leptin changes are explained by the concomitant insulin variation should be further studied under renal failure conditions. PMID- 9734620 TI - Cyclosporine microemulsion increases drug exposure and reduces acute rejection without incremental toxicity in de novo renal transplantation. International Sandimmun Neoral Study Group. AB - BACKGROUND: The new oral microemulsion formulation of cyclosporine (Neoral) possesses superior pharmacokinetics to the conventional formulation, Sandimmun (SIM), providing more complete and predictable absorption, and less pharmacokinetic variability. METHODS: The safety and tolerability of Neoral, together with the incidence of acute rejection episodes and graft survival, were compared to the conventional cyclosporine formulation, SIM, in a prospective, randomized, double-blind multicenter trial. A total of 167 patients who received a first or second cadaveric renal transplant in 21 participating centers in six countries were randomized equally to two treatment groups and followed for three months after transplantation. Outcomes were analyzed across treatment, center and regional groups. In addition, a nested pharmacokinetic study was performed in four of these centers throughout the period of follow-up. RESULTS: No difference was detected between the safety or tolerability of the two formulations. Kidney function and other laboratory parameters remained comparable in Neoral- and SIM treated patients throughout the study. However, the number of patients experiencing acute rejection was significantly reduced for the Neoral group (44.2% vs. 60.5%; P = 0.044), and significantly fewer patients experienced multiple episodes of rejection (12.8% vs. 22.2%, P = 0.028). The proportion of patients free of rejection at three months was significantly higher in patients treated with Neoral than in those receiving SIM (Kaplan-Meier estimated probability of remaining rejection-free at 3 months = 55% for the Neoral group, compared with 39% for the SIM group, P = 0.046, log rank test). Similar results were obtained when acute rejection, graft loss and death were used as a combined endpoint (Kaplan-Meier estimated probability for Neoral group = 54%, compared with 38% for the SIM group, P = 0.047, log rank test). Comparison of results by center or regional groups did not show any significant treatment interaction. A nested pharmacokinetic evaluation (four centers; 28 subjects) showed that the bioavailability of cyclosporine from Neoral was significantly higher than from SIM at all assessment times. Specifically, at weeks 2, 4 to 6, and 12, dose normalized AUC was 49%, 63% and 32% higher for Neoral. Dose-normalized peak cyclosporine blood concentrations and AUC stabilized by weeks 4 to 6 in patients receiving Neoral, whereas these values increased slowly in SIM-treated patients without reaching the levels achieved in the Neoral group. CONCLUSIONS: These results suggest that the superior pharmacokinetic characteristics of the microemulsion formulation of cyclosporine lead to more efficient immunosuppression during the first critical months after transplantation, without a deleterious impact on clinical safety. PMID- 9734621 TI - CO2 and gadopentetate dimeglumine as alternative contrast agents for malfunctioning dialysis grafts and fistulas. AB - BACKGROUND: Hemodialysis grafts and native fistulas are frequently evaluated angiographically utilizing iodinated contrast material to determine the cause of malfunction. Occasionally, patients are not able to receive iodinated contrast material due to a history of previous severe allergic reaction or concern that iodinated contrast material could worsen renal function requiring premature initiation of permanent dialysis. We set out to test the feasibility of gadopentetate dimeglumine as an alternative contrast agent in conjunction with carbon dioxide (CO2) angiography in the evaluation and treatment of hemodialysis grafts and native fistulas in patients who have a contraindication to iodinated contrast material. METHODS: Six patients with a malfunctioning hemodialysis graft and native fistula were evaluated. Four patients were successfully evaluated using carbon dioxide and gadopentetate dimeglumine. Two additional patients underwent balloon angioplasty using gadopentetate dimeglumine alone as the alternative contrast agent. RESULTS: All six patients successfully were evaluated and treated using gadopentetate dimeglumine either alone or as a supplement to CO2 angiography. Five of these patients had lesions successfully treated using gadopentetate dimeglumine alone or in combination with CO2 as the angiographic contrast agents. One patient underwent a successful diagnostic angiogram using gadopentetate dimeglumine and CO2 as alternative contrast agents and was subsequently treated with surgical revision. The gadopentetate dimeglumine angiograms identified the arterial anastomosis and more clearly identified stenotic lesions and venous outflow anatomy compared to carbon dioxide angiograms. CONCLUSION: Gadopentetate dimeglumine is useful as an alternative contrast agent in conjunction with CO2 in patients with malfunctioning hemodialysis grafts and fistulas, who have a contraindication to the administration of iodinated contrast material. PMID- 9734622 TI - Body composition in children receiving recombinant human growth hormone after renal transplantation. AB - BACKGROUND: Recombinant human growth hormone (rhGH) is an anabolic hormone promoting protein synthesis in various tissues. Therefore, changes in body composition may be expected during rhGH treatment. METHODS: We studied changes in body composition during two years of rhGH treatment in 21 children after at least one year with a functioning renal transplant. The mean +/- SD age was 12.9+/-2.5 years at the start of rhGH therapy. A whole body, dual energy X-ray absorptiometry (DEXA) exam was performed before the initiation of rhGH therapy (T0), and was repeated at one and two year intervals after initiation of the therapy (T1 and T2, respectively). RESULTS: Lean body mass increased by a median of 0.48 SDS during the first year of treatment (P = 0.022), and the median increase during two years of therapy was 0.36 SDS (P = 0.061). On the contrary, the median fat body mass decreased by 2.17 SDS during the T0 to T1 period (P = 0.04) and by 1.99 SDS during the T0 to T2 period (P = 0.055). The index for fat body mass/lean body mass (FBM/LBM) decreased by a median of 5.3% during T0 to T1 (P < 0.001), however, a slower but still significant decrease by a median of 4.2% was noted at T2 (P < 0.05). Bone mass content did not change significantly during rhGH treatment. The medians in caloric and protein intakes were stable during rhGH treatment. CONCLUSION: A significant increase of lean body mass and a decrease of fat body mass was noted during rhGH therapy in children after renal transplantation. PMID- 9734623 TI - Prevalence of histological beta2-microglobulin amyloidosis in CAPD patients compared with hemodialysis patients. AB - BACKGROUND: The prevalence of beta2-microglobulin amyloidosis (Abeta2m) in patients on continuous ambulatory peritoneal dialysis (CAPD) is unknown. METHODS: We prospectively obtained a median of 2 (range 1 to 4) joint samples from 26 CAPD patients aged 44 to 93 (median 73) years at post-mortem evaluation after 4.5 to 126 (median 27) months solely on CAPD (N = 19) or primarily on CAPD (that is, < or = 10% and < or = 1 year of renal replacement therapy time on other modalities; N = 7). The diagnosis of Abeta2m rested on Congo red staining (typical birefringence) and positive immunostaining of amyloid deposits by a monoclonal anti-beta2m antibody. RESULTS: Abeta2m was diagnosed in 8 of 26 patients (31%). Prevalence ranged from 20% (2 of 10 patients) within < or = 24 months CAPD to 30% (3 of 10 patients) after 24 to 48 months and 50% (3 of 6 patients) after 49 to 126 months (P = 0.11). The prevalence of Abeta2m was similar in patients without or with one or more peritonitis episodes. No significant difference in prevalence (P = 0.118) was found between CAPD patients (8+/26; 31%) and hemodialysis patients (13+/26; 50%) carefully matched for time on dialysis and age at the onset of dialysis. CONCLUSIONS: The prevalence of histological Abeta2m reaches 31% after a median duration of 27 months of CAPD. This prevalence is not significantly different from that observed in a group of HD patients matched for age and dialysis duration. PMID- 9734624 TI - Elevated neointimal endothelin-1 in transplantation-associated arteriosclerosis of renal allograft recipients. AB - BACKGROUND: Chronic renal allograft rejection is characterized histologically by transplantation-associated arteriosclerosis and glomerulosclerosis (Tx-AA and Tx AGS). Recent studies in animal models implicate the mitogenic and pressor actions of endothelin-1 (ET-1) in Tx-AA. In humans, however, a link between elevated ET-1 secretion and Tx-AA or Tx-AGS remains unclear. In this study we analyzed expression of ET-1 in the vasculature of renal transplant patients with chronic or acute rejection and in normal controls. METHODS: Renal vascular and glomerular ET-1 was assessed by immunohistochemistry in 12 patients with clinically and histologically defined chronic rejection, in 11 patients with acute rejection, and in 5 normal kidneys. ET-1 staining was also correlated with various clinical parameters and with a morphometric index of neointima formation. ET-1 secretion was measured by ELISA in cultured human vascular cell types treated with T cell- and macrophage-associated cytokines. RESULTS: We found that renal allografts with chronic rejection and Tx-AA expressed 6.1-fold more ET-1 in the vasculature relative to allografts with acute rejection or to normal kidneys (P < 0.01). In Tx-AA, ET-1 was detected predominantly in the neointima, which contained mostly endothelial cells and smooth muscle cells. A strong positive correlation (r = 0.82, P < 0.01) was observed between vascular ET-1 peptide expression and hypertension in patients with chronic rejection. We also showed that macrophage associated cytokines, but not T cell-associated cytokines, stimulated ET-1 secretion in human endothelial cells, vascular smooth muscle and mesangial cells. CONCLUSIONS: These results demonstrate that elevated ET-1 in the neointima is associated with Tx-AA and chronic rejection. In addition, these results point to an important role for endothelial dysfunction in chronic renal allograft rejection. PMID- 9734625 TI - Delayed graft function of more than six days strongly decreases long-term survival of transplanted kidneys. AB - BACKGROUND: We reviewed 843 first cadaver kidney transplants carried out consecutively at our center to examine the effect on long-term graft survival of the duration of delayed graft function (DGF), defined as the time taken for the kidney to attain the threshold of a Cockcroft calculated creatinine clearance (cCCr) > or = 10 ml/min. METHODS: Using a multivariate Cox survival analysis we evaluated the consequences of DGF on allograft survival, and then by regression analysis identified the factors contributing to the occurrence of DGF. Finally, using a Kaplan Meier analysis we compared the profiles of graft failure according to the duration of DGF. RESULTS: Defining DGF in terms of cCCr rather than necessity for dialysis after transplantation allowed better prediction of long term graft loss. Indeed, patients with a Cockcroft-based DGF > six days who did not require dialysis (12%) had a significantly poorer long-term graft outcome than those with a DGF < or = six days. Furthermore, we showed that a DGF of six days could be taken as a cut-off point that marked a significant difference in the long-term graft survival rate (P < 0.0001). Surprisingly, further extension of the duration of DGF > six days was not associated with further worsening of graft survival (except in DGF > 30 days). CONCLUSION: Our results suggest a threshold effect in the lesions that ultimately results in long-term functional deficiency. In addition, we show that the need for dialysis is not an adequate criterium for DGF in terms of long-term outcome prediction. PMID- 9734626 TI - Enhancement of convective transport by internal filtration in a modified experimental hemodialyzer: technical note. AB - BACKGROUND: Hemodialysis using high flux membranes today is a commonly used therapy. The primary advantage is the larger spectrum of molecules removed with these membranes, and the mechanism of removal is in part due to a phenomenon of filtration and backfiltration along the length of the hollow fibers. We hypothesized that increasing the filtration and backfiltration fluxes by modifying the structure of the dialyzer could enhance the convective transport of various solutes. METHODS: A modified high flux dialyzer was compared to the standard model in terms of pressure profiles, filtration-backfiltration rates and solute clearances. The modification consisted on the placement of a O-ring around the fiber bundle to create a resistance for the flow of the dialysis solution external to the fibers. The study on filtration fluxes was carried out using a scintigraphic method previously described, and solute clearances were studied during ultrafiltration-controlled hemodialysis sessions. RESULTS: Utilizing a net filtration condition proximal to zero, the rates of proximal filtration and distal backfiltration in the experimental dialyzer were significantly enhanced in comparison with the standard dialyzer. The pressure drop in the dialysate compartment could be increased significantly, thus permitting an increase in the positive transmembrane pressure in the first half of the dialyzer and a parallel increase in the negative transmembrane pressure in the second half of the dialyzer. This resulted in a significant enhancement of the convective transport of middle-large solutes as demonstrated by the increase in vitamin B12 and inulin clearances. CONCLUSIONS: This approach suggests that changes in design of the dialyzer may affect its performance. The use of internal filtration is suggested to improve convection and dialyzer efficiency for larger solutes without the requirement for high volumes of replacement fluid, as is the case for current hemodiafiltration techniques. PMID- 9734627 TI - Use of iohexol to quantify hemodialysis delivered and residual renal function: technical note. AB - BACKGROUND: Classically, urea (molecular wt = 60) is used to determine the urea reduction ratio (URR) or clearance, based on volume of distribution (Kt/V). These methods are subject to many errors. The purpose of this study was to determine whether iohexol (Io; molecular wt = 821) could be used instead of urea and provide better information as well as middle molecule clearance data. METHODS: Ten hemodialysis (HD) patients were evaluated. All were dialyzed for three hours, and a single bolus of 100 ml of Io was injected immediately post-HD. For direct dialysis quantification (DDQ), the spent dialysate was collected in a drum, and urea and iodine (I) determined immediately prior to, at the end of, and 30 minutes post-HD. As routinely used, DDQ measures clearance directly rather than estimates the levels. RESULTS: Calculated Kt/V urea (1.21+/-0.05) significantly overestimated DDQ Kt/V urea (0.78+/-0.04, P < 0.001) whereas calculated and DDQ Kt/V Io were similar (1.44+/-0.10 vs. 1.36+/-0.05). The URR and iohexol reduction ratio (IoRR) were also different (0.63+/-0.02 vs. 0.69+/-0.02; P < 0.002) with a urea but not Io rebound (URR30 min 0.59+/-0.02, P < 0.05). Calculated urea clearance (C(urea)), 247+/-21 ml/min, significantly overestimated DDQ C(urea) (157+/-10 ml/min P < 0.001). Calculated CIo and DDQ CIo, however, were similar (109+/-8 vs. 104+/-7 ml/min). Total body clearance (TBC) in six anuric subjects was 2.5+/-0.3 ml/min, and in four oliguric subjects was 5.2+/-0.5 ml/min. In 10 additional patients, direct urine measurements demonstrated a non-renal clearance (NRC) of 2.97+/-0.18 ml/min, which was 4.0+/-0.3% of body wt. Use of this factor allowed an estimation of residual renal function (RRF) that accurately reflected measured RRF (1.32+/-0.53 vs. 1.42+/-0.55 ml/min) CONCLUSION: A single injection of Io can be used to determine Kt/V, RR, and RRF without rebound or the inconvenience of urine collection. It may also represent middle molecule clearance better than urea kinetics, and may serve as a superior method for determining HD delivered and dialysis adequacy. PMID- 9734628 TI - The tubulointerstitium in progressive renal disease. PMID- 9734629 TI - Do oxidized lipoproteins contribute to glomerulosclerosis? PMID- 9734630 TI - Uremic hyperleptinemia: adaptive or maladaptive? PMID- 9734631 TI - Malnutrition and dialysis. PMID- 9734632 TI - Hypokinetic azotemic osteodystrophy. PMID- 9734633 TI - Toward global advancement of medicine: the International Society of Nephrology experience. AB - BACKGROUND: Since its foundation in 1960, the International Society of Nephrology (ISN) has pursued the worldwide advancement of education, science and patient care in nephrology. This goal was achieved by means of the Society's journal and the organization of international congresses and symposia. In order to better reach its colleagues and patients in economically less developed countries, the ISN expanded its activities as of 1980 by a large number of specific programs aimed at these regions. METHODS: The first phase of activities included teaching programs, fellowship and visiting scholar programs, and the provision of travel grants to enhance accessibility to the ISN congresses. A second phase consisted of the creation of a library enhancement program, a commission on acute renal failure and--to improve the organization and efficiency--a central commission on global advancement of nephrology (COMGAN). Currently, a third phase has been entered in which all activities have been intensified: (1) under the guidance of COMGAN, supported by a large number of teaching programs and fact finding missions; (2) by establishing a renal sister program; and (3) by initiating commissions on informatics and on clinical trials. RESULTS: As a result, the ISN has reached most parts of the world, previously deprived of contact with renal science and renal patient care. The fellowship program now counts 160 fellows, who spend one or two years in training. The library enhancement program reaches 218 institutions worldwide. ISN membership has soared over the past two years with over 2,500 new members, mostly in the developing countries. They receive Kidney International and other relevant forms of information. Thus far, 135 pairs of renal units in developing and developed countries have been linked for support on a more continuous basis. ISN-sponsored congresses, symposia, and courses are being held in increasing numbers in the developing world. In many of its activities, the ISN closely collaborates with sister organizations, which also contribute financially. In total, the ISN spends annually over $1 million US from its own budget on the programs described above. CONCLUSION: The various programs and initiatives are proving helpful in advancing renal medicine in areas in need. Expansion into supporting similar programs within other medical subspecialties is being explored. PMID- 9734634 TI - Planned progressive antimicrobial therapy in neutropenic patients. PMID- 9734635 TI - Elevated TFPI in malignant disease: relation to cancer type and hypercoagulation. AB - We have previously reported high levels of the coagulation inhibitor TFPI in the blood of patients with gastrointestinal cancer. TFPI is not an acute-phase reactant, but high levels have also been reported in patients with septicaemia and disseminated intravascular coagulation (DIC). To study its relationship with other types of malignancy, TFPI activity was first determined in plasma samples from 214 patients with various malignancies. In a second cohort of 83 patients, total and free TFPI antigen, protein C, antithrombin, fibrin monomer and D-dimer were also measured. Elevated TFPI activity and antigens were found in about half of the patients with solid tumours. In contrast, elevated TFPI was rare in haematological malignancies (12%). In the 18 patients with acute nonlymphocytic leukaemia (ANLL), elevated free TFPI was found only in patients who also had DIC. No correlation was found between TFPI levels and fibrin monomer or D-dimer levels. Only four out of 20 patients with solid tumours had normal levels of fibrin monomer and D-dimer, yet three out of these four had elevated TFPI. In conclusion, elevated TFPI in ANLL is related to the coexistence of DIC. In solid tumour disease increased TFPI may reduce protective fibrin formation, but the pathogenic mechanism is as yet unknown. PMID- 9734636 TI - Antibodies to prothrombin in antiphospholipid syndrome and inflammatory disorders. AB - Antiphospholipid antibodies associated with the antiphospholipid syndrome (APS) have been shown to bind plasma proteins, particularly beta 2-glycoprotein I (beta2-GPI). In this study the incidence of antibodies to solid-phase prothrombin was examined in patients with antiphospholipid syndrome and a variety of other inflammatory disorders. Significantly elevated levels of IgG anti-prothrombin (anti-PT) antibodies were detected in 63% of patients with APS (n = 27, median 22 arbitrary units: AU), 33% with SLE (n = 92, median 14 AU). 45% with rheumatoid factor (n = 22, median 16 AU), 21% with carotid artery stenosis (n = 21, median 15 AU), 32% with stroke (n = 38, median 13 AU). 67% of patients with a false positive serology for syphilis (n = 21, median 24 AU), 37% with HIV (n = 30, median 14 AU), 29% with syphilis (n = 14, median 19 AU) and 3% with infectious mononucleosis (n= 30, median 9 AU). In addition, a group of lupus anticoagulant (LA) positive patients (n = 48) was examined for antibodies to prothrombin, beta2 GPI and cardiolipin. 10 (21%) patients had raised levels of IgG anti-PT antibodies, 30 (62%) had significantly elevated levels of anti-beta2-GPI antibodies and 15 (31%) had elevated levels of anticardiolipin antibodies (ACA). Of the LA-positive patients, 15 (43%) were identified with definite APS, eight (23%) with probable APS, two (6%) with possible APS and 10 (28%) patients had no clinical evidence of APS. In conclusion, antibodies to prothrombin were found in a variety of inflammatory disorders and were therefore not specific for the APS. However, identification of the plasma proteins recognized by antibodies from patients with APS may provide insight into the pathogenic mechanisms involved in the heterogenous clinical manifestations of the APS. PMID- 9734637 TI - Lupus anticoagulant, anticardiolipin antibodies and hepatitis C virus infection in thalassaemia. AB - Anticardiolipin antibodies (ACA) and lupus anticoagulant (LA) have been detected in patients with hepatitis C virus (HCV) infection and have been associated in autoimmune diseases (i.e. systemic lupus erythematosus) with an increased risk of thromboembolic events. Because of the high prevalence of HCV infection and the thrombotic risk described in thalassaemia we decided to investigate the prevalence of ACA and LA in a cohort of 68 thalassaemia patients. We found a high prevalence (34%) of beta2-glycoprotein I independent ACA in our thalassaemia patients which was related to HCV infection. None of patients developed any complications related to antiphospholipid antibodies (APL); therefore the clinical significance of positivity for APL in patients with HCV infection is at present unclear. In conclusion, the results of our study indicate that ACA in the serum of HCV-infected thalassaemic patients exhibit the characteristics of natural autoantibodies rather than those of the pathogenic autoantibodies that are found in patients with systemic lupus erythematosus. PMID- 9734638 TI - Review of computerized decision support systems for oral anticoagulation management. AB - Computerized decision support systems (CDSS) are available to assist clinicians in the therapeutic management of oral anticoagulation. We report the findings relating to CDSS for oral anticoagulation management of a primary-care-based systematic review which largely focused on near-patient testing. Seven papers were reviewed which covered four different systems. The methodology of these papers was generally poor, although one randomized controlled trial showed improved therapeutic control associated with computerized management compared with human performance. PMID- 9734639 TI - A comparison of linear and orthogonal regression analysis for local INR determination in ECAA coagulometer studies. European Concerted Action on Anticoagulation. AB - International sensitivity index calibrations based on the W.H.O. recommended method depend on orthogonal regression analysis. As this is not readily available in statistical packages, comparison has been made with simple linear regression analysis in a study of coagulometer effects on the International Normalized Ratio (INR) at 155 European centres. Sets of seven lyophilized normal and 20 lyophilized artificially depleted abnormal plasmas were provided with five coumarin test plasmas and two European Concerted Action on Anticoagulation reference thromboplastins (low International Sensitivity Index (ISI) human and high ISI rabbit). Local ISI based on the artificially depleted lyophilized plasmas using conventional orthogonal regression gave good correction for local coagulometer effects on the human reagent and minimal correction with the rabbit reagent INR. Results were considerably worse after attempts at correction using calibration based on linear regression analysis with both reagents. The results indicate that calibration of coagulometer prothrombin time systems using simple linear regression is not appropriate. PMID- 9734640 TI - Double heterozygosity of the GPIIb gene in a Swiss patient with Glanzmann's thrombasthenia. AB - Glanzmann's thrombasthenia (GT) results from a qualitative or quantitative defect of GPIIb-IIIa complexes (integrin alphaIIbbeta3). the fibrinogen receptor on platelets. This integrin plays a critical role in platelet aggregation. In this report we describe the molecular abnormalities of a patient with clinical and laboratory findings typical of type I Glanzmann's thrombasthenia. SDS-PAGE with Western blotting revealed an absence of GPIIb but small amounts of normally migrating GPIIIa in his platelets. A non-radioactive PCR-SSCP procedure and direct sequence analysis of PCR-amplified DNA fragments showed the patient to be a compound heterozygote for mutations in the GPIIb gene. A single point mutation (G to A) at nucleotide 1064 of the cDNA derived from the mother's allele led to a Glu324 to Lys amino acid substitution in GPIIb. It was responsible for a MscI restriction site in exon 12 of the GPIIb gene. This amino acid substitution changes the electric charge between the second and third Ca++-binding domains of GPIIb. The second mutation was inherited from his father and is in exon 18 of the GPIIb gene. It was a T --> C base transition at position 1787 of GPIIb cDNA and results in a Ile565 to Thr substitution. The two GPIIb mutations identified in this study will provide new information on GPIIb-IIIa structure and biosynthesis. PMID- 9734641 TI - Molecular abnormality observed in a patient with coagulation factor X (FX) deficiency: a novel three-base-pair (CTT) deletion within the polypyrimidine tract of the FX intron D. AB - We investigated the molecular defect underlying congenital factor X (FX) deficiency in a Japanese patient. A novel three-base-pair deletion was identified within intron D of the FX gene. An allele-specific restriction analysis showed the propositus was homozygous and her two daughters were heterozygous for the deletion. It was not detected in 53 unrelated Japanese (106 alleles), indicating a probable cause of the FX deficiency. The deletion resides within a polypyrimidine tract of the acceptor splicing site where U2 snRNP binds to form spliceosomes. The defect could alter the formation of spliceosomes, resulting in incorrect splicing and decreased FX production. PMID- 9734642 TI - Budd-Chiari syndrome, portal vein and mesenteric vein thrombosis in a patient homozygous for factor V Leiden mutation treated by TIPS and thrombolysis. AB - We present the first case of Budd-Chiari syndrome in association with portal and mesenteric vein thrombosis in a patient homozygous for the factor V Leiden mutation. She was treated by transjugular intrahepatic portosystemic stent (TIPS) placement followed by local thrombolytic therapy. Venous outflow from the liver was established and the thrombi in the portal and mesenteric veins were lysed completely. This therapeutic approach may be used for such patients with this severe thrombotic event, who generally have a poor prognosis. PMID- 9734644 TI - Factor binding to the human gamma-globin gene distal CCAAT site: candidates for repression of the normal gene or activation of HPFH mutants. AB - We have examined factor binding to the distal human gamma-globin CCAAT site and three naturally occurring hereditary persistence of fetal haemoglobin (HPFH) mutations of this site. Factor binding was examined using nuclear extracts from the erythroleukaemic cell lines K562 and MEL, and from A4 cells, a non transformed mouse bone marrow stem cell line, using the electrophoretic mobility shift assay. Under standard binding conditions, in addition to the previously reported binding by a CCAAT factor (CP1) and GATA-1, the wild-type (wt) sequence bound high mobility factors which appeared to be GATA-2 isoforms. However, when the non-specific competitor conditions were varied, the binding profile with K562, but not MEL nuclear extract, was substantially altered. CP1 and GATA-1 were absent, and two new factors were detected, one of which bound preferentially to the Greek and Japanese non-deletion HPFH mutants. However, binding by the GATA-2 isoforms to the wt sequence was maintained with both cell types, as it was using the A4 cell line. With modified binding conditions, in A4 cells the two non deletion and the Black deletion HPFH mutants each had a different protein binding profile which was lost on erythroid induction of the cells. We discuss the possibility that the GATA-2 isoforms bound to the wt sequence may function to suppress wt gamma gene expression in the bone marrow. Additionally, those factors which bind preferentially either to the deletion or non-deletion HPFH mutants may play positive roles in establishing an active chromatin structure. PMID- 9734643 TI - Homozygous missense mutation (band 3 Fukuoka: G130R): a mild form of hereditary spherocytosis with near-normal band 3 content and minimal changes of membrane ultrastructure despite moderate protein 4.2 deficiency. AB - The characteristics of phenotypic expression were studied in a Japanese family with hereditary spherocytosis and an extremely rare homozygous missense mutation of the band 3 gene (band 3 Fukuoka: G130R). The homozygous unsplenectomized proband was a 29-year-old male with compensated haemolytic anaemia (red cell count 4.21 x 10(12)/l, reticulocytes 278 x 10(9)/l, and indirect bilirubin 44 micromol/l). His red cell band 3 (B3) protein demonstrated a 9.3% reduction and his protein 4.2 (P4.2) level was substantially reduced (45.0%), compared to normal subjects. P4.2 protein was composed mostly of a wild type (72 kD) with a trace of 68 kD peptide. The binding properties of the mutated B3 to normal P4.2 were significantly impaired, which probably resulted in the substantial reduction of P4.2 in this proband, since no abnormalities were detected on the P4.2 gene. Electron microscopy (EM) using the freeze-fracture method demonstrated a mild decrease in intramembrane particles (IMPs) of near-normal size (8 nm in diameter) with no substantial increases in their oligomerization. Their distribution on the membrane P face was almost normal, although most of the IMPs could represent the homozygously mutated B3 protein. EM (quick-freeze deep-etching method) disclosed a skeletal network of near-normal size and size distribution of the skeletal units, suggesting that the mutated B3 protein itself did not have much effect on the skeletal network in situ. Therefore the reduced P4.2 content (45% of that of normal subjects), which remained on the red cell membrane of this proband, appeared to be nearly sufficient for maintaining the normal structure of the skeletal network and IMPs in situ, contrary to the marked abnormalities in both IMPs and the skeletal network in complete P4.2 deficiencies. PMID- 9734645 TI - Leucocyte filterability: comparing diluted with undiluted blood. AB - Red blood cells and about 95% of white blood cells have an immediate and constant effect on the flow of undiluted or diluted blood through 5 microm filters. The remaining 5% of all leucocytes exert an increasing influence on flow such that the rate of flow of diluted and undiluted blood through these filters is continually declining over a period of 150 s. Analysis of this declining flow rate enables these cells to be counted and their rheological properties to be deduced. Approximately 50% of these slow leucocytes pass through the filters with a transit time of about 30 s and the remaining cells act as pore blockers for 150 s. The numbers and flow properties of slow leucocytes was approximately the same in blood from young women (25 years) and older men (65 years). However, the number of slow leucocytes was increased in a group of men (65 years) suffering from peripheral arterial occlusive disease. Dilution of the blood with phosphate buffered saline increased the numbers of slow leucocytes in both of the older, but not the younger, group of volunteers. This effect was particularly noticeable in the patient group. It is recommended that filtration studies of the rheological profile of leucocytes can, and must, be performed with undiluted blood. The properties after dilution may sometimes, but not invariably, reflect changes ex vivo as well as inherent differences in the cells themselves. PMID- 9734646 TI - Senescent human neutrophil binding to thrombospondin (TSP): evidence for a TSP independent pathway of phagocytosis by macrophages. AB - This study investigated the interaction of apoptotic polymorphonuclear neutrophils (PMN) with thrombospondin (TSP), an important event mediating the clearance of apoptotic neutrophils by macrophages. We developed an in vitro assay to examine this interaction. Based on this assay, we found that apoptotic but not fresh PMN bound specifically to surface-immobilized TSP (33 +/- 0.03 x 10(3) cells/well) compared to fibrinogen, fibronectin or laminin (8.0 +/- 0.3 x 10(3) cells/well). Moreover, the binding was specific for surface bound but not soluble TSP and appeared to be divalent cation dependent, was not significantly inhibited by heparin and was sensitive to cycloheximide (CHX) treatment of senescent PMN (>90%) inhibition at 10 microM CHX). In contrast to the binding studies, phagocytosis of senescent PMN by macrophages was not affected by EDTA or cycloheximide. Phosphatidyl-L-serine liposomes, phospho-L-serine, glucosamine, galactosamine, and the acetylated sugars had no effect on phagocytosis. We conclude that: (i) there was specific binding of senescent human PMN to immobilized TSP, which is divalent cation dependent and requires new protein synthesis in the PMN during senescence; (ii) in addition to the recently defined TSP-dependent pathway, there is a TSP-independent pathway mediating phagocytosis of senescent PMN by macrophages. The identity of this pathway remains to be defined. PMID- 9734647 TI - A novel surface marker (B203.13) of human haemopoietic progenitors, preferentially expressed along the B and myeloid lineages. AB - We used a monoclonal antibody (mAb) (B203.13, IgM) generated from a mouse immunized with the human B/myeloid bi-phenotypic B1b cell line, to analyse haemopoietic cells. The antigen recognized by this mAb is expressed on most adult and umbilical cord blood CD21+ B cells, at minimal density on mature monocytes, and is undetectable on granulocytes, T, natural killer (NK) cells, and erythrocytes. Within umbilical cord blood and adult bone marrow haemopoietic progenitor cells, the B203.13 mAb recognized a surface marker, present on progenitor cells of several haemopoietic lineages, that was transiently expressed on early erythroid and T/NK progenitors, and was preferentially maintained on cells of the B and myeloid lineages. Within the CD34+ cells, B203.13 was expressed on early committed myeloid (CD33+) and erythroid (CD71dim) progenitor cells, as confirmed in colony formation assays. The mAb also reacted with cells of B and myeloid chronic leukaemias and cell lines. These data define B203.13 mAb as a novel reagent useful for the characterization of haemopoietic progenitors and leukaemias. PMID- 9734648 TI - Bb1-3, a transgenic hybrid cell line with erythroid and megakaryocytic differentiation potential that expresses high levels of human gamma-globin and human beta-globin. AB - We have characterized a murine hybrid cell line, Bb1-3, generated by the fusion of mouse primary erythroblasts with MEL cells. It proliferated in serum-free medium and displayed a low level of spontaneous erythroid and megakaryocyte differentiation. Terminal erythroid differentiation could be induced with HMBA and DMSO and was enhanced by serum. Treatment with phorbol esters resulted in a high proportion of megakaryocytes and the expression of megakaryocytic specific lineage markers. Bb1-3 cells contain a human beta-globin transgene that was expressed at levels of 20-50% of the endogenous mouse globin genes. Initially, expression was largely limited to the beta-globin gene but after adaptation to serum free growth, equal expression of both the human gamma- and human beta globin genes was observed. This cell line provides further evidence that the differentiation potential of mouse erythroleukaemia cells is not restricted to the erythroid lineage and should be useful to study the mechanisms underlying both developmental globin gene regulation and the terminal differentiation of bipotential erythroid/megakaryocytic progenitor cells. PMID- 9734649 TI - Molecular cloning and functional analysis of the CD33 promoter. AB - CD33 is a leucocyte differentiation antigen restricted to myeloid cells in blood and bone marrow. Two mRNA transcripts encoding CD33 are observed in leukaemic cell lines. The smaller transcript of 1.5 kb is comparable in size to the isolated CD33 cDNA but the origin of the larger 1.8 kb transcript is unknown. To study the regulation of human CD33 expression, a 5' genomic clone from the CD33 gene was isolated and studied for promoter activity. The clone, although lacking a TATAA box, exhibits other sequences characteristic of a promoter. Two transcriptional start sites were identified, 414 and 527 nucleotides 5' of the ATG initiation codon, suggesting that these sites are used to generate the 1.8 kb transcript observed in CD33+ cell lines. The CD33 genomic sequence directed high expression of a luciferase reporter gene in myeloid cell lines. Using deletion mutants of the promoter sequence, maximal expression was localized to the first 220 bp 5' of the ATG initiation codon. Site-directed mutagenesis of an Sp-1 and PU.1 binding site within this region showed that the PU.1, but not Sp-1, was critical for CD33 expression in myeloid lines. Given the restricted expression of CD33 on haemopoietic cells, the identification of the CD33 promoter may be useful for the study of transcription factors that regulate gene expression during early myeloid differentiation. PMID- 9734650 TI - Unbalanced X-chromosome inactivation in haemopoietic cells from normal women. AB - We studied X-chromosome inactivation patterns in blood cells from normal females in three age groups: neonates (umbilical cord blood), 25-32 years old (young women group) and >75 years old (elderly women). Using PCR, the differential allele methylation status was evaluated on active and inactive X chromosomes at the human androgen receptor (HUMARA) and phosphoglycerate kinase (PGK) loci. A cleavage ratio (CR) > or = 3.0 was adopted as a cut-off to discriminate between balanced and unbalanced X-chromosome inactivation. In adult women this analysis was also performed on hair bulbs. The frequency of skewed X-inactivation in polymorphonuclear (PMN) cells increased with age: CR > or = 3.0 was found in 3/36 cord blood samples, 5/30 young women and 14/31 elderly women. Mathematical analysis of patterns found in neonates indicated that X-chromosome inactivation probably occurs when the total number of haemopoietic stem cell precursors is 14 16. The inactivation patterns found in T lymphocytes were significantly related to those observed in PMNs in both young (P < 0.001) and elderly women (P < 0.01). However, the use of T lymphocytes as a control tissue for distinguishing between skewed inactivation and clonal proliferation proved to be reliable in young females, but not in elderly women, where overestimation of the frequency of clonal myelopoiesis may appear. PMID- 9734651 TI - Long-term detection of microchimaerism in peripheral blood after pretransplantation blood transfusion. AB - Renal allograft survival is prolonged after pretransplantation blood transfusion. The aim of this study was to test retrospectively the development and persistence of microchimaerism after pretransplantation blood transfusion and to assess whether the type of blood transfusion (partially matched [= sharing of at least one HLA-B and one HLA-DR antigen between blood donor and recipient] versus mismatched) influences the (continued) presence of donor-type cells. A sensitive nested PCR technique based on HLA-DRB1 allele-specific amplification using sequence-specific primers (detection level: one donor cell among 10(5) recipient cells) for detection of donor cells was implemented in our laboratory. We studied 21 patients for microchimaerism in the peripheral blood compartment, following blood transfusion. Our preliminary data show that microchimaerism was detectable up to 8 weeks after blood transfusion. In all patients receiving a partially matched blood transfusion, donor-type cells were detected in the first week after transfusion, in 7/8 patients 2-4 weeks after transfusion, and in some patients up to 8 weeks after transfusion. After mismatched transfusion a tendency to shorter duration of microchimaerism was observed. PMID- 9734652 TI - Genomic typing of the Kidd blood group locus by a single-tube allele-specific primer PCR technique. AB - The Kidd (JK) blood group system is clinically important in transfusion medicine. Alloantibodies to antigens in this system may be produced following blood transfusion or during pregnancy and can result in serious haemolytic transfusion reactions and haemolytic disease of the newborn (HDN). JK antigens on erythrocytes are carried by glycoproteins with the capacity to transport urea through cell membranes. cDNA complementary to mRNA transcribed at the JK locus was cloned in 1994. The molecular basis of the Jk(a)/Jk(b) blood group polymorphism was recently shown to be a single nucleotide substitution predicting an amino acid change (Asp280Asn) in an extracellular loop of the JK glycoprotein. After confirmation of the JK gene polymorphism we developed a rapid and robust technique for JK genotyping with allele-specific primers in a single-tube PCR. In addition, a 217 bp intron located at nucleotides 811-812 in the JK gene was found and sequenced. The genotyping test was validated with samples from 106 Caucasian Swedish and 13 Black South African random blood donors. Complete phenotype genotype correlations were obtained. However, four Jk(a-b-) samples of Polynesian and Finnish origin typed as Jk(b)Jk(b). Potential use of the presented method can be predicted in clinical transfusion medicine including prenatal determination of the JK genotype in a fetus at risk for HDN caused by JK antibodies. PMID- 9734653 TI - Quinine improves the results of intensive chemotherapy in myelodysplastic syndromes expressing P glycoprotein: results of a randomized study. AB - Intensive chemotherapy produces a lower complete remission (CR) rate in the myelodysplastic syndromes (MDS) than in de novo acute myeloid leukaemia (AML), possibly due in part to a higher incidence of P glycoprotein (PGP) expression in MDS blast cells. We designed a randomized trial of intensive chemotherapy with or without quinine, an agent capable of reverting the multidrug resistance (mdr) phenotype, in patients aged < or = 65 years with high-risk MDS. Patients were randomized to receive mitoxantrone 12 mg/m2/d days 2-5 + AraC 1 g/m2/12 h days 1 5, with (Q+) or without (Q-) quinine (30 mg/kg/d). 131 patients were included. PGP expression analysis was successful in 91 patients. In the 42 PGP-positive cases, 13/25 (52%) patients in the Q+ group achieved CR, compared to 3/17 (18%) patients in the Q- group (P = 0.02) and median Kaplan-Meier survival was 13 months in the Q+ group, and 8 months in the Q- group (P = 0.01). No life threatening toxicity was observed with quinine. In conclusion, the results of this randomized study show that quinine increases the CR rate and survival in PGP positive MDS cases treated with intensive chemotherapy. PMID- 9734654 TI - Serum levels of p55 and p75 soluble TNF receptors in adult acute leukaemia at diagnosis: correlation with clinical and biological features and outcome. AB - The tumour necrosis factor (TNF)/TNF-receptor (TNFR) complex plays a role in the growth of leukaemic cells. We retrospectively investigated the relationship between pretreatment serum concentration of soluble TNFR (p55- and p75-sTNFRs) and outcome in adult acute myeloid (AML 82 cases) and lymphoid (ALL 44 cases) leukaemia. Both sTNFRs were significantly higher in AML (p55-sTNFR 4.53 +/- 3.7, median 3.75; p75-sTNFR 6.51 +/- 5.25 ng/ml, median 4.72) and ALL sera (3.31 +/- 1.5, median 2.95; 5.30 +/- 2.3 ng/ml, median 4.56, respectively) than in controls (1.89 +/- 0.5, median 1.98; 2.22 +/- 0.8 ng/ml, median 2.37) (P < 0.01 for both sTNFRs). Fresh leukaemic cells expressed p55- and p75-sTNFRs, which were modulated and released into the supernatant (SN) following short-term in vitro culture, suggesting that in vivo sTNFRs were also leukaemia-derived. Whereas no correlation was observed between sTNFRs and outcome in ALL, in AML higher p55 sTNFR levels (> 3.75 ng/ml) were associated with shorter disease-free survival (DFS) (P = 0.006) and overall survival (OS) (P = 0.0004). At multivariate analysis p55-sTNFR was the most significant predictor of DFS (P = 0.006) and OS (P < 0.001). Our data suggest that the prognostic significance of p55-sTNFR in AML could be related to relevant biological features of AML blasts. PMID- 9734655 TI - Immunophenotype of adult and childhood acute promyelocytic leukaemia: correlation with morphology, type of PML gene breakpoint and clinical outcome. A cooperative Italian study on 196 cases. AB - Acute promyelocytic leukaemia (APL), characterized by a specific PML-RARalpha fusion gene resulting from translocation t(15;17) and by a high response rate to differentiation therapy with all-trans retinoic acid, presents clinical (varying WBC counts, age and treatment outcome), morphological (hypergranular M3 and hypogranular M3V) and molecular (three isoforms of PML breakpoint) heterogeneity. We correlated leukaemic immunophenotype with these aspects in 196 molecularly confirmed APLs (63 children and 133 adults) in Italy. The bcr3 isoform (P = 0.05) and FAB M3V (P = 0.05) were more frequent in children. We confirmed in APL an immunophenotype characterized by frequent expression of CD13, CD33 and CD9 and rare expression of HLA-DR, CD10, CD7 and CD11b. However, we recognized CD2 in 28%, CD34 in 23% and CD19 in 11% of cases and demonstrated by double labelling that CD34 and CD2 may be co-expressed. CD2, CD34 and CD19 were significantly intercorrelated, and variably associated to other features: CD2 and CD34 with PML bcr3 (P < 0.001 and P < 0.001, respectively) and with M3V (P < 0.001 and P = 0.002), whereas only CD19 was directly correlated with WBC counts and only CD2 positively influenced CR rate (logistic model) and event-free survival (Cox model). We conclude that immunophenotype plays a role in the determination of the biological and clinical heterogeneity of childhood and adult APL. PMID- 9734656 TI - Inhibited apoptosis and drug resistance in acute myeloid leukaemia. AB - Despite extensive investigation into mechanisms of drug resistance in acute myeloid leukaemia (AML), the aetiology of therapeutic resistance is unclear. We found that five leukaemia cell lines (K562, HL-60, CEM. CEM induced to overexpress bcl-2, and REH) displayed parallel sensitivity to four antileukaemia drugs with different mechanisms of action, with K562 generally being the least sensitive and REH being the most sensitive. The amount of spontaneous apoptosis in the cell lines after serum-free culture paralleled their drug sensitivity: K562 cells displayed the least apoptosis at 24h (2.50 +/- 0.24%) and REH the most (24.47 +/- 8.22%). The extent of spontaneous apoptosis of leukaemic blasts from 39 patients with newly diagnosed de novo AML also correlated with the success of the intensive, infusional cytarabine-based induction therapy. There was a median of 19.5% (range 3.6-64%) apoptotic AML cells after 24 h of serum-free culture in patients who entered a complete remission compared with 4.2% (1.8-7.0%) apoptotic AML cells in patients who did not achieve a complete remission (P = 0.0007). Thus, inhibited apoptosis was associated with both in vitro and in vivo pan resistance to antileukaemic chemotherapy. The cause of inhibited apoptosis in AML is probably a function of interactions among multiple signals that influence apoptosis. Assessment of spontaneous apoptosis may serve as an important prognostic factor for AML. PMID- 9734657 TI - Multiplex PCR for TCR delta rearrangements: a rapid and specific approach for the detection and identification of immature and mature rearrangements in ALL. AB - The preferential occurrence of immature T-cell receptor (TCR) delta rearrangements (i.e. incomplete Ddelta2-Ddelta3 and Vdelta2-Ddelta3) in B-cell precursor acute lymphoblastic leukaemia (BCP ALL) and of predominantly mature rearrangements (incomplete Ddelta2-Jdelta1, complete Vdelta1, Vdelta2, Vdelta3 to Jdelta1) in T-lineage ALL prompted us to establish two separate multiplex PCR systems for the identification of clonal TCRdelta rearrangements. PCR products of the expected size for the specific rearrangements were detectable from a dilution of 100-1000 clonal cells in 150000 polyclonal cells. Both multiplex PCR systems were used to analyse samples from 86 childhood BCPALLs and 30 T-lineage ALLs. The results of the multiplex PCRs were controlled by standard PCR analyses for the individual rearrangements and Southern blots, which were identical. Only immature TCRdelta rearrangements were detected in BCP ALL (59%), whereas no rearrangement was found in the remaining BCP leukaemias, thus confirming the exclusive presence of immature TCRdelta rearrangements in B-lineage cells. 50% of the T-lineage ALLs contained mature rearrangements, but no immature rearrangements were found. These two multiplex PCR techniques appear to be reliable and fast aids in the analysis of clonal TCRdelta rearrangements in ALL. PMID- 9734658 TI - Arsenic induces apoptosis in B-cell leukaemic cell lines in vitro: activation of caspases and down-regulation of Bcl-2 protein. AB - We showed that arsenic inhibited the cell growth of four B-cell leukaemia cell lines of 11 various cell lines in vitro. In two of these four lines, KOCL44 and LyH7, apoptosis was identified by morphological and nucleosomal DNA fragmentation studies. Three of the four B-cell lines that were growth inhibited were acute infantile leukaemia with t(11;19)(q23;p13) translocations involving the MLL gene that encodes the transcriptional factor Drosophila trithorax. The arsenic-induced apoptosis in KOCL44 and LyH7 cells was found to be linked to caspases by Western blot and enzymological analyses. The amount of Bcl-2 was reduced during apoptosis in LyH7 as judged by Western blot analysis. We concluded that combined activation of the caspases and down-regulation of Bcl-2 could determine the fate of B-cell leukaemic cells in response to arsenic. PMID- 9734659 TI - Targeting of saporin to Hodgkin's lymphoma cells by anti-CD30 and anti-CD25 bispecific antibodies. AB - CD25 and CD30 represent suitable target molecules for bispecific antibody (bimAb) driven toxin delivery to lymphoid tumour cells. We describe two new anti CD30/anti-saporin bimAbs (termed CD30 x sap1 and CD30 x sap2), produced by hybrid hybridomas, which react against non-cross-reactive epitopes of the saporin molecule, and compared their effect with a bimAb reacting with saporin and with CD25 (CD25 x sap1). In a protein synthesis inhibition assay these bimAbs were able to enhance saporin toxicity (IC50 = 8.5 x 10(-9) M in the absence of mAbs) with a similar activity: in the presence of 10(-9) M CD30 x sap1 bimAb the IC50 was 2.75 x 10(-11) M, whereas with CD30 x sap2 bimAb the IC50 was 6.5 x 10(-11) M and CD25 x sap1 bimAb displayed an IC50 of 3 x 10(-11) M (as saporin). The combined use of the two anti-CD30 bimAbs further increased cytotoxicity by 100 fold, resulting in an IC50 of 1.9 x 10(-13) M. A slightly less efficient improvement was obtained by combining the CD25 x sap1 bimAb with the CD30 x sap2 bimAb directed against a different toxin epitope (saporin IC50 to 7 x 10(-13) M). In contrast, no synergistic effect was observed using the combination of the anti CD25 bimAb with the anti-CD30 bimAb reacting with the same epitope of saporin (IC50 = 4.5 x 10(-11) M). Analysis of FITC-saporin binding to L540 cells by flow cytometry demonstrated that the appropriate combinations of the two anti CD30/anti-saporin bimAbs or of the anti-CD30/anti-saporin and anti-CD25/anti saporin bimAbs had a cooperative effect on the binding of the ribosome inactivating protein (RIP) to the cells, when compared with single bimAbs. PMID- 9734660 TI - Functional granulocyte/macrophage colony stimulating factor receptor is constitutively expressed on neoplastic plasma cells and mediates tumour cell longevity. AB - It has been shown that granulocyte/macrophage colony stimulating factor (GM-CSF) is able to support myeloma cell propagation in cooperation with interleukin (IL) 6, the major growth factor for malignant plasma cells, although the biological mechanisms involved remain unknown. Therefore we investigated (i) the expression levels of the GM-CSF receptor (GM-CSFR) constituents in three malignant plasma cell lines and in native malignant plasma cells, (ii) the ability of the receptor to mediate common signalling pathways regulating proliferation and cell survival in malignant plasma cell lines, and (iii) the effects of GM-CSF on tumour cell biology. The GM-CSFRalpha subunit was detected in the malignant plasma cell lines RPMI-8226, MC/CAR, IM-9 as well as 6/6 native myeloma cell samples derived from the bone marrow of patients with overt disease. Furthermore, GM-CSFR expression was also detected in the CD19+ fraction from 2/3 bone marrow samples and 5/8 peripheral blood samples derived from patients with malignant plasma cell disorders, but not in the CD19+ fraction of peripheral blood from healthy donors. The expressed cytokine receptor alpha-subunit was able to constitute a functional signalling complex with the ubiquitously expressed GM-CSFRbeta subunit, as demonstrated by the fact that GM-CSF induced the p21-ras/mitogen-activated protein kinase (MAPK) signalling cascade in malignant plasma cell lines. Since this signalling cascade plays an essential role in the mediation of both proliferation and cell survival, we investigated the impact of GM-CSF on these two events. Application of GM-CSF led to an increase of DNA-synthesis in MC/CAR, IM-9 and RPMI-8226 cells. Furthermore, it increased longevity of these malignant plasma cell lines by reducing the rates of spontaneous apoptosis. We conclude that (i) the functional GM-CSFR is commonly expressed on malignant plasma cells and that (ii) GM-CSF promotes the clonal expansion of myeloma cells by inhibiting spontaneous apoptosis and promoting DNA synthesis. PMID- 9734661 TI - Establishment and characterization of EBV-positive and EBV-negative primary effusion lymphoma cell lines harbouring human herpesvirus type-8. AB - In this study we report on the establishment and characterization of two novel lymphoma cell lines (CRO-AP/3 and CRO-AP/5) which carry infection by human herpesvirus type-8 (HHV-8) and have derived from AIDS-related primary effusion lymphoma (PEL). These two cell lines are representative of different virologic subtypes of PEL, i.e. HHV-8+/EBV- PEL in the case of CRO-AP/3 and HHV-8+/EBV+ PEL in the case of CRO-AP/5. Consistent with the diagnosis of PEL, both CRO-AP/3 and CRO-AP/5 expressed indeterminate (i.e. non-B, non-T) phenotypes although immunogenotypic studies documented their B-cell origin. Both cell lines are devoid of genetic lesions of c-MYC, BCL-2 and p53 as well as gross rearrangements of BCL-6. Detailed histogenetic characterization of these novel PEL cell lines suggests that PEL may derive from a post-germinal centre B cell which has undergone pre-terminal differentiation. The CRO-AP/3 and CRO-AP/5 cell lines may provide a valuable model for clarifying the pathogenesis of PEL. In particular, these cell lines may help understand the relative contribution of HHV-8 and EBV to PEL growth and development and may facilitate the identification of recurrent cytogenetic abnormalities highlighting putative novel cancer related loci relevant to PEL. PMID- 9734662 TI - Dexamethasone plus retinoids decrease IL-6/IL-6 receptor and induce apoptosis in myeloma cells. AB - Interleukin 6 (IL-6) is the most important known growth factor for multiple myeloma, and IL-6 signalling pathways are potential targets for therapy. We hypothesized that interfering with the IL-6 signalling pathway at more than one level would be more effective than a single block in inhibiting proliferation of myeloma cells. Accumulating data support the concept that glucocorticoids down regulate IL-6, whereas retinoic acid derivatives (RA) down-regulate IL-6R in myeloma. We found that all-trans RA (ATRA), 13-cis-RA and 9-cis-RA each similarly inhibited growth of RPMI 8226 myeloma cells and that addition of dexamethasone (DEX) added to RA growth inhibition. The major effects of retinoids were to reduce the proliferative fraction and induce apoptosis whereas DEX increased the apoptotic fraction. When combined, apoptosis was enhanced. Effects of RA + DEX were also least able to be overcome by exogenous IL-6. RA decreased IL-6R levels and addition of DEX to RA delayed recovery of IL-6R levels compared with RA alone. Since RPMI 8226 cells have undetectable IL-6, we investigated U266B1 cells and found that RA and DEX decreased both IL-6 secretion and IL-6 RNA levels. Mechanistically, IL-6R down-regulation by RA was enhanced by DEX, whereas IL-6 protein and RNA levels were reduced by DEX and by RA. In summary, combinations of RA + DEX were not only more effective in inhibiting myeloma cells growth by the dual mechanisms of decreasing proliferative fraction and increasing apoptotic fraction, but were also less able to be overcome by IL-6. PMID- 9734663 TI - Thiamine-responsive myelodysplasia. AB - The triad of thiamine-responsive anaemia, diabetes mellitus and deafness has been reported in 15 patients with macrocytic anaemia, sometimes associated with moderate thrombocytopenia. The bone marrow aspirate usually shows megaloblastic changes and ringed sideroblasts. However, tri-lineage myelodysplasia has never been reported. We describe two patients who presented with diabetes, deafness and thiamine-responsive pancytopenia. Bone marrow aspirate and biopsy were typical of tri-lineage myelodysplasia. These findings suggest that thiamine may have a role in the regulation of haemopoiesis at the stem cell level. We propose the term 'thiamine-responsive myelodysplasia' rather than that of thiamine-responsive anaemia. PMID- 9734665 TI - Consensus statement of the Royal College of Physicians of Edinburgh Consensus Conference on Medical Management of Stroke, 26-27 May 1998. PMID- 9734664 TI - Mobilizing peripheral blood stem cells with high-dose G-CSF alone is as effective as with Dexa-BEAM plus G-CSF in lymphoma patients. AB - We compared retrospectively the efficacy of granulocyte colony stimulating factor (G-CSF) alone with chemotherapy plus G-CSF in mobilizing CD34-positive cells in patients with malignant lymphoma. 35 patients underwent peripheral blood stem cell (PBSC) collection following mobilization either with 24 microg/kg G-CSF for 4 consecutive days (n = 18) or Dexa-BEAM chemotherapy plus 5 microg/kg G-CSF (n = 17). High-dose G-CSF was well tolerated with only slight bone pain and/or myalgia. The Dexa-BEAM therapy required hospitalization with a median duration of 21 d. The median number of apheresis procedures in both groups was two (range two to four), resulting in a median of 5.3 and 5.1 x 10(6) CD34+ cells/kg. No patients in the G-CSF group, but one in the Dexa-BEAM group, failed to reach the target of collecting >2.0 x 10(6) CD34+ cells/kg. The number of CFU-GM (10.4 v 6.0 x 10(5)/kg) and of BFU-E (10.6 v 4.5 x 10(5)/kg; P = 0.04) was higher in the G-CSF group than in the Dexa-BEAM group. A subset analysis of CD34+ cells was performed in 16 patients showing a higher mean of Thy-1 (CD90w) coexpression in the G-CSF than in the Dexa-BEAM group (4.8 v 1.8%, P = 0.12). Additionally the percentage of CD34+/CD38- cells was higher in the G-CSF group (10.66% v 8.8%). However, these differences were not statistically significant. The median time to leucocyte and platelet engraftment after high-dose chemotherapy was slightly shorter in the G-CSF than in the Dexa-BEAM group (9 v 10 and 12 v 13.5 d, respectively). These results demonstrate that high-dose G-CSF is as effective as Dexa-BEAM plus G-CSF in mobilizing peripheral blood stem cells and produces prompt engraftment. The major advantages of G-CSF mobilization were the safe outpatient self-application and the fixed-day apheresis. PMID- 9734666 TI - Anaphylactic reaction to liposomal amphotericin (AmBisome) PMID- 9734667 TI - The use of amphotericin B lipid complex in 15 patients with presumed or proven fungal infection. PMID- 9734668 TI - A warfarin induction regimen for out-patient anticoagulation in patients with atrial fibrillation. PMID- 9734669 TI - Severe juvenile haemochromatosis (JH) missing HFE gene variants: implications for a second gene locus leading to iron overload. PMID- 9734670 TI - In patients with lymphoid tumours recovering from the autoimmune complications of fludarabine, relapse may be triggered by conventional chemotherapy. PMID- 9734671 TI - Workers' compensation and chronic regional musculoskeletal pain. PMID- 9734672 TI - Genetic studies of common rheumatological diseases. PMID- 9734673 TI - Azathioprine and cyclophosphamide in the treatment of rheumatoid arthritis. PMID- 9734674 TI - Direct cost of rheumatoid arthritis during the first six years: a cost-of-illness study. AB - The objective was to estimate the annual direct disease-related cost of rheumatoid arthritis (RA) during the first 6 yr and to determine which socio demographic and clinical characteristics relate to these costs. The study population consisted of 424 RA patients who had participated in a (population based) trial on therapeutic strategies for early RA since 1990 and were not lost to follow-up in April 1996. A questionnaire on costs due to RA was sent to these patients; 363 (86%) completed questionnaires were analysed. The total annual direct cost per patient was estimated by adding up the costs of health care workers, days admitted to care facilities, medication, monitoring for side effects, alternative medicine, adaptations in the home, devices, and other direct costs such as travelling expenses. The mean annual direct cost due to RA was estimated to be Dfl. 11,550 per patient. An obvious increase in direct cost with increasing disease duration was not found. Patients with higher disease activity exhibited significantly higher costs compared to patients with lower disease activity. A multiple logistic regression model showed that greater disability and lower age increased the odds for high costs. The annual direct cost of RA averaged out at Dfl. 11,550 per patient (i.e. Pound Sterling 3680). A high total direct cost in the first 6 yr of disease is related to severe functional disability and lower age. PMID- 9734675 TI - Effects of rheumatoid arthritis on employment and social participation during the first years of disease in The Netherlands. AB - OBJECTIVE: To study the effect of rheumatoid arthritis (RA) on working capabilities and social participation, including non-paying jobs, during the first 6 yr of disease. DESIGN: Cross-sectional study. METHODS: In April 1996, a self-reporting questionnaire was sent to 424 participants of a population-based clinical trial of therapeutic strategies for early RA initiated in 1990. RESULTS: A total of 363 completed questionnaires were returned (response = 86%). Disease duration varied from < 1 to 6 yr (mean 2.8 yr). The employment rate was low in the RA population compared to the Dutch population. In the male 45- to 64-yr-old group, 63% of RA patients were not employed compared to 32% of the Dutch population (P < 0.01). In the female 45- to 64-yr-old group, 76% of the RA population vs 67% of the Dutch were not employed (P < 0.05). Of the employed patients, 59% reported that RA affected their working capabilities, e.g. they worked an average of 21 h per week less due to RA. Of the patients without a paying job, 41% believed that this was (partly) due to RA. In addition, fewer RA patients had non-paying jobs and they performed fewer household activities compared to the general Dutch population. CONCLUSION: RA already has a negative influence on the working capabilities, social participation and household activities of these patients during the first 6 yr of disease. PMID- 9734676 TI - Assessment of disease activity in rheumatoid arthritis using magnetic resonance imaging: quantification of pannus volume in the hands. AB - We attempted to assess whether pannus volume measured by magnetic resonance imaging (MRI) can be used as an indicator of disease activity in rheumatoid arthritis (RA). Eleven women (mean age 46 yr) with uncontrolled RA were studied for 1 yr. Pannus formation in both hands was quantified using MRI at the start of the study, and at 6 and 12 months thereafter. The volume of enhancing pannus (VEP) was compared with changes in the radiological scores, grip strength, joint tenderness counts, joint swelling counts, erythrocyte sedimentation rate (ESR), and serum C-reactive protein (CRP). Patients were classified into three groups based on VEP changes between 0 and 12 months: unchanged (n = 2), decreased (n = 6) and increased (n = 3). VEP at 6 months and at 12 months differed significantly between the three groups. No statistically significant differences were found between the groups in radiographic scores, physical parameters or laboratory parameters despite the fact that some of these parameters changed in the direction indicated by the changes in VEP. VEP can be used as a new indicator to assess disease activity in individual RA patients and, using this parameter, treatment outcome can be assessed in fewer subjects than with traditional measures. PMID- 9734677 TI - Comparison of the MOS short form-12 (SF12) health status questionnaire with the SF36 in patients with rheumatoid arthritis. AB - OBJECTIVE: To compare the performance of the MOS SF12 health survey (SF12) with the SF36 in a sample of 233 patients with rheumatoid arthritis (RA) stratified by functional class. METHODS: The SF12 and SF36 physical and mental component summary scales (PCS and MCS) were compared for test retest reliability [intra class correlation coefficient (RC) and repeatability], construct validity and responsiveness [standardized response mean (SRM)] to self-reported change in health. RESULTS: Overall, despite its brevity, the SF12 is comparable to the SF36 with only some loss of performance. The SF12-PCS is slightly less reliable (RC = 0.75) and responsive to improvements in health (SRM = 0.52) than the SF36-PCS (RC = 0.81; SRM = 0.61). The SF12-PCS correlates strongly with the SF36-PCS (R = 0.94), SF36 physical function subscale (R = 0.77) and modified Stanford Health Assessment Questionnaire (MHAQ) (R = 0.71), but only weakly with the SF36 mental health subscale (R = 0.22). SF12-PCS discriminated well between Steinbrocker functional classes; patients in functional classes 1-4, respectively, have SF12 PCS scores 1sigma, 2sigma, 2.4sigma and 2.7sigma below the population norm (ANOVA, F = 35.8, P < 0.000). The SF12-MCS is relatively unresponsive to reported improvement in RA (SRM = 0.31), but is reliable (RC = 0.71) and correlates well with the SF36-MCS (R = 0.71). SF12-MCS correlates more closely than the SF36-MCS with the SF36 mental health subscale (R = 0.86) and Hospital Anxiety and Depression (HAD) scale (R = 0.76). In ANOVA models, only the HAD (R2 = 57%) score contributes significantly to variance in SF12-MCS (F = 254.8; P < 0.000), but both the HAD (R2 = 24%) and MHAQ (R2 = 10%) scores contribute to variance in the SF36-MCS (F = 50.9; P < 0.000). Thus, the SF12-MCS has better construct validity for mental health than SF36-MCS in RA subjects. Missing responses to items were high amongst patients in functional class 4 (34%). CONCLUSION: The SF12 is a reliable, valid and responsive measure of health status in the majority of RA patients, and meets standards required for comparing groups of patients. Its application in the most severely disabled subjects is uncertain. PMID- 9734678 TI - Knee pain and disability in the Nottingham community: association with poor health status and psychological distress. AB - OBJECTIVE: To assess the prevalence of knee pain, disability and health status in the community, and to examine the association of pain with psychological distress. METHODS: A postal survey was sent to 4057 men and women aged 40-79 yr in Nottingham. Health status was assessed using the SF-36 instrument, with the specific dimensions of physical function and mental health used to measure disability and psychological distress. RESULTS: The overall response rate was 81.9%. The prevalence of knee pain was 28.7%, rising with age. Disability was more common in those with knee pain compared to those without pain (P < 0.001). Subjects with knee pain had lower scores for all dimensions of health. When adjusted for potential confounders, low mental health scores associated with increased odds for pain and disability (2.1, 95% CI 1.7-2.6; and 4.7, 95% CI 3.7 6.1). CONCLUSIONS: Knee pain is common in this population and is associated with poor perceived health and significant disability. Psychological distress strongly associates with both pain and disability. PMID- 9734679 TI - Progression of joint damage in early active severe rheumatoid arthritis during 18 months of treatment: comparison of low-dose cyclosporin and parenteral gold. AB - OBJECTIVE: This study compared the progression of joint damage in patients with early active severe rheumatoid arthritis (RA) treated with cyclosporin or parenteral gold. METHODS: In this open, randomized, multicentre study with a blinded radiological endpoint, 375 patients who had suffered from active severe RA for <3 yr were randomized to be treated for 18 months with low-dose cyclosporin or parenteral gold. The groups were stratified with regard to corticosteroid use. Primary efficacy variables were numbers of erosions, erosion score and the Larsen-Dale joint damage score. RESULTS: Joint damage progressed at similar rates in both treatment arms. In both groups, patients receiving corticosteroids had less X-ray progression. Rheumatoid factor positivity, high swollen joint count, high erythrocyte sedimentation rate and pre-existing X-ray abnormalities predicted progression of joint damage. Although numbers of serious adverse events were similar, more gold patients (n = 65) than cyclosporin patients (n = 45) withdrew from study medication because of adverse events. CONCLUSION: Cyclosporin was comparable to parenteral gold in retarding progression of joint damage and was better tolerated in terms of adherence to therapy. The open label design should be kept in mind when assessing this difference. PMID- 9734680 TI - Arterial disease in lupus and secondary antiphospholipid syndrome: association with anti-beta2-glycoprotein I antibodies but not with antibodies against oxidized low-density lipoprotein. AB - The prevalence and clinical significance of antibodies against beta2-glycoprotein I (anti-beta2GPI) and antibodies against oxidized low-density lipoprotein (anti ox-LDL) were evaluated as potential indicators of arterial disease in patients with systemic lupus erythematosus (SLE) and SLE with secondary antiphospholipid syndrome (APS). IgG anti-beta2GPI and IgG anti-ox-LDL were measured by enzyme linked immunosorbent assay (ELISA) in serum samples from 118 patients with SLE, including 40 with secondary APS. IgG anti-beta2GPI were positive in 17% (20/118) of SLE patients. The presence and titres of IgG anti-beta2GPI were strongly associated with a history of arterial thrombosis. Haemolytic anaemia was also significantly associated with the presence of IgG anti-beta2GPI. The prevalence of IgG anti-ox-LDL was 53% (63/118), but there was no association with arterial thrombosis. No correlation between the values of anti-ox-LDL and those of anti beta2GPI was found. These results suggest that IgG anti-beta2GPI could be a marker for arterial thrombosis in SLE patients, while IgG anti-ox-LDL were not associated with arterial disease in this group of lupus patients. PMID- 9734681 TI - Sleep apnoea caused by rheumatoid arthritis. AB - Sleep apnoea syndrome (SAS) is a rarely documented, but possibly lethal, complication of the instability of the cervical spine in rheumatoid arthritis. Five patients with SAS of a central or peripheral origin are presented, and the problems of recognizing and diagnosing the syndrome are discussed. We hope that clinicians will become more aware of the existence and the different aetiologies of SAS, thus improving early recognition and appropriate treatment. Adequate treatment has proven to increase survival in peripheral SAS and seems to be successful in doing so in central SAS. PMID- 9734682 TI - Risk factors for avascular bone necrosis in systemic lupus erythematosus. AB - OBJECTIVE: To study the predictive factors for avascular necrosis (AVN) of bone in patients with systemic lupus erythematosus (SLE). METHOD: The records of 38 SLE patients who developed clinically apparent AVN during the course of their disease were reviewed. Information on clinical presentation, corticosteroid usage and autoantibody profiles was obtained, and comparison was made between these patients and 143 consecutive control SLE patients who did not have AVN. RESULTS: The point prevalence of AVN in our SLE population was 12%. Patients with AVN, when compared with controls, had a significantly higher incidence of neurological disease (39% vs 14%; P < 0.001) and Cushingoid body habitus after steroid treatment (79% vs 53%; P = 0.004). The highest cumulative prednisolone dose in 1 and 4 months was significantly higher in the AVN group than the controls (1.8 vs 1.1 and 4.5 vs 2.8 g, respectively; P < 0.01 in both) and showed a linear trend with the incidence of AVN (chi2 test for trend, P < 0.01 in both). Lupus anticoagulant was associated with AVN (P = 0.02, odds ratio 2.88 [1.14-7.28]). Logistic regression analysis revealed that the highest cumulative prednisolone dose administered in 4 months, the maximum and mean daily prednisolone dosage, and the lupus anticoagulant were independent risk factors for AVN. CONCLUSIONS: Corticosteroid remains the major predisposing factor for AVN in SLE. Patients who require an initial high-dose steroid for disease control are at risk of AVN, especially if they are positive for the lupus anticoagulant or develop Cushingoid habitus after steroid treatment. High-risk patients should be closely monitored so that early AVN can be diagnosed by sensitive techniques such as magnetic resonance imaging and radioisotope bone scanning. PMID- 9734683 TI - Leucocyte membrane expression of proteinase 3 correlates with disease activity in patients with Wegener's granulomatosis. AB - Wegener's granulomatosis (WG) is an inflammatory disorder characterized by granulomatous inflammation and vasculitis, and is strongly associated with antineutrophil cytoplasmic antibodies (ANCA). ANCA in patients with WG are directed against proteinase 3 (Pr3) in most of the cases. In vitro, upon neutrophil priming, ANCA antigens are expressed on the cell surface, thereby becoming available for interaction with ANCA. Subsequently, these neutrophils become activated. Since ANCA can only interact with leucocytes when the ANCA antigens are present on the cell surface, we questioned whether Pr3 is already expressed on the membranes of circulating granulocytes and monocytes of patients with WG, and whether Pr3 expression is related to disease activity, so explaining the systemic nature and severity of the disease. The expression of Pr3, and other ANCA antigens, i.e. myeloperoxidase (MPO) and human leucocyte elastase (HLE), was analysed on circulating granulocytes and monocytes by flow cytometry, using a non activating whole-blood method. Disease activity was quantitated using the Birmingham Vasculitis Activity Score (BVAS). Seventeen patients with active WG and anti-Pr3 antibodies were included in this study. Nine of these patients were also analysed at the time of remission. Twelve patients with sepsis served as positive controls, and 10 healthy volunteers as negative controls for granulocyte/monocyte activation. Pr3 expression on neutrophils was increased in patients with active WG compared to patients with quiescent disease and healthy controls. On monocytes, no differences in Pr3 expression were found between those groups. Furthermore, the expression of MPO and HLE did not differ between patient groups and healthy controls. Upon follow-up, the expression of Pr3 on neutrophils from patients with active WG decreased when patients went into remission. Pr3 expression on neutrophils correlated with the BVAS score (r = 0.40, P < 0.05). In conclusion, circulating neutrophils from patients with active WG have increased expression of Pr3. In addition, the expression of Pr3 correlates with disease activity, suggesting that the availability of Pr3 for interaction with ANCA plays a central role in the disease process. PMID- 9734684 TI - Methotrexate treatment in Felty's syndrome. AB - Felty's syndrome is a rare disorder characterized as a systemic manifestation of severe rheumatoid arthritis associated with granulocytopenia and splenomegaly. We report a retrospective analysis of a series of seven patients treated successfully with low-dose methotrexate. leading to sustained clinical improvement (number of swollen joints) and normalization of the granulocyte count for an observation period of 1 yr. Our cohort is the largest ever published with methotrexate treatment of this rare condition. Our results confirm earlier single case reports suggesting methotrexate to be the first-choice treatment nowadays in Felty's syndrome. PMID- 9734685 TI - Modification of the inflammatory activity of psoriatic arthritis in patients treated with extract of Polipodium leucotomos (Anapsos) PMID- 9734686 TI - An unusual case of carpal tunnel syndrome. PMID- 9734687 TI - Tumour markers in dermatomyositis: useful or useless? PMID- 9734688 TI - Serious opportunistic infection associated with gold-induced panhypogammaglobulinaemia. PMID- 9734690 TI - Chondrodysplasic rheumatism. PMID- 9734689 TI - Necrotizing arteritis confined to striated muscle mimicking a soft-tissue tumour in a patient with rheumatoid arthritis. PMID- 9734691 TI - Meeting highlights: therapeutic strategies and supportive care in myelodysplastic syndromes: yesterday, today and tomorrow. PMID- 9734692 TI - Risk factors and their relationship to prognosis in myelodysplastic syndromes. AB - Recent efforts have been directed at improving the methodology for predicting clinical outcomes in patients with myelodysplastic syndromes (MDS). This review focuses on the development of a consensual, prognostic, risk-based analysis system generated by the International MDS Risk Analysis Workshop. In the workshop, cytogenetic, morphological, and clinical data were combined and collated from a relatively large group of patients with primary MDS. Critical prognostic variables were evaluated using the data set. Based on these findings, the International Prognostic Scoring System (IPSS) was developed, compared with other systems, and shown to provide more accurate prognoses regarding survival and evolution to acute myeloid leukemia in MDS patients. The improvement was due to several features of the workshop model: more refined cytogenetic categorization, inclusion of cytopenias, improved subdivision of marrow blast percentages, four subgroups defining outcome, and separate stratification for age. The IPSS should result in better-defined clinical outcomes in MDS and provide a framework for future studies determining the possible role of molecular determinants (e.g. oncogenes, tumor suppressor genes, cytokine expression and responsiveness) for evaluating prognoses. The IPSS will likely prove useful in the design and analysis of therapeutic trials in MDS as well as in patient management. PMID- 9734693 TI - Hematopoietic stimulation by amifostine and sodium phenylbutyrate: what is the potential in MDS? PMID- 9734694 TI - Erythropoietin, with and without granulocyte-colony stimulating factor (G-CSF), in the treatment of myelodysplastic syndrome (MDS) patients. PMID- 9734695 TI - Standard and low-dose chemotherapy for the treatment of myelodysplastic syndromes. AB - The myelodysplastic syndromes (MDS) are a heterogeneous group of disorders with an invariably fatal outcome. Other than bone marrow transplantation, no treatment has been able to alter the natural history of MDS. As a result, there has been an interest in identifying new and more effective chemotherapeutic agents. Many of the drugs that have been evaluated in an attempt to increase remissions and prolong survival were selected because of their activity in acute myeloid leukemia. Thus cytarabine has been the most widely studied drug. Although numerous studies suggested activity for low-dose cytarabine (LoDAC), a careful analysis of the data identified a complete remission (CR) rate of less than 20%, without meaningful clinical benefit. The issue of LoDAC was finally put to rest by a randomized trial in which survival was no better than with supportive care. 5-Azacytidine induces cellular differentiation by hypomethylation of DNA. Phase II trials noted CRs in fewer than 10% of patients, with response rates under 30%, although additional patients appeared to experience hematologic and clinical benefit. A randomized trial of 5-azacytidine versus supportive care failed to demonstrate a survival benefit. One of the most promising new agents is the topoisomerase inhibitor topotecan, which achieves CRs in more than 30% of patients. Combinations of this drug with other active agents are in development. Obviously, new treatment strategies are needed to improve the outcome of MDS patients. Combination approaches incorporating new, active agents should have sound scientific rationale, targeting biological differences among the various MDS subtypes. PMID- 9734696 TI - Intensive therapy for high-risk myelodysplastic syndromes and the biological significance of karyotype abnormalities. AB - Therapy for myelodysplastic syndromes (MDS) has been less than effective when based on low-dose treatment or supportive measures only, including hematopoietic growth factors. Recently, based on the percentage of bone marrow blasts, the number of cytopenic cell lines and cytogenetics, clinical risk groups have been defined more precisely. Recent studies applying intensive acute myeloid leukemia (AML)-type therapy to high-risk MDS have produced remissions ranging from 45 to 79%. Advances in the understanding of the biology of MDS clearly point to cytogenetics rather than morphologic subtype as being of prognostic relevance. Hence, new treatments need to be developed for patients with unfavorable karyotypes and complex abnormalities in particular. These MDS subtypes are characterized by low spontaneous proliferative activity and low autocrine production of hematopoietic growth factors. The subtypes are, however, highly sensitive to external stimulation by granulocyte-colony stimulating factor (G CSF) and granulocyte macrophage-colony stimulating factor (GM-CSF). New therapies could emerge from these findings, for example, priming high-risk MDS patients with hematopoietic growth factors in combination with intensive AML-type treatment. Recent studies suggest that incorporating high-dose AraC into an intensive drug combination could further improve the outcome of high-risk MDS. PMID- 9734697 TI - Reducing the toxicity of anticancer therapy: new strategies. AB - Cytoprotective agents offer opportunities to reduce the treatment-related toxicity of anticancer therapy and perhaps to increase the dose and dose intensity of radiation and chemotherapy. One such agent is amifostine, an organic thiophosphate. Amifostine selectively protects normal tissues and provides broad spectrum protection for a variety of organs while remaining minimally toxic. Clinical studies have demonstrated that amifostine protects against myelotoxicity, nephrotoxicity, neurotoxicity, mucositis and esophagitis in patients treated with alkylating and platinum agents, paclitaxel and radiation therapy. In addition, preclinical studies suggest the possibility of protection against anthracycline-induced cardiotoxicity and radiation- and chemotherapy induced mutagenicity. Preclinical and clinical studies have not demonstrated any diminution of antitumor efficacy. Amifostine is well tolerated in doses of 740 or 910 mg/m2. The most common side effects requiring treatment are transient hypotension, which responds to intravenous fluids, and nausea and vomiting, effectively treated with 5-HT3 antagonists and dexamethasone. PMID- 9734698 TI - Bone marrow transplantation for myelodysplasia in adults and children: when and who? PMID- 9734699 TI - Quality of life and psychosocial adjustment in patients with myelodysplastic syndromes. AB - Health-related quality of life (QOL) is a dynamic, subjective, multidimensional concept. In chronic illnesses such as myelodysplastic syndromes (MDS), management typically focuses on control of the disease and its symptoms. Yet an equally important consideration is maintaining the patient's QOL. Much research remains to be done on the subject of QOL in the MDS population. MDS patients have unique problems that can affect QOL: they tend to be elderly and to have comorbid conditions; they develop complications resulting from cytopenias such as infection and bleeding; they experience fatigue; they run the risk of converting to a form of acute leukemia typically resistant to therapy; and they face uncertainty. This paper will address the above problems using a conceptual approach to QOL that has been useful for patients with cancer and various chronic illnesses. The approach, based on the work of Ferrell and Cella, concentrates on five areas of the patient's life: the physical, functional, emotional, social, and spiritual. Actual testing of this conceptual framework in the MDS patient population has begun. Here methods of systematically testing and assessing QOL are discussed in order to help clinicians meet the primary purpose of therapy in the setting of chronic illness: improving or preserving the individual's QOL. PMID- 9734700 TI - Stability and distribution of orally administered epidermal growth factor in neonatal pigs. AB - Stability and distribution of orally administered epidermal growth factor (EGF) were examined in newborn and 5-day-old pigs. Forty-five minutes after oral administration of iodine-125 labeled EGF, 60 and 50% of the radioactivity administered were recovered from the internal organs in newborn and 5-day-old pigs, respectively. In both age groups, over 95% of the recovered radioactivity was found in the gastrointestinal tract, of which 78-86% was found in the luminal contents with the remaining found in the gastrointestinal wall. Within the gastrointestinal tract, 65-71% of radioactivity was found in the stomach, 27-30% in the proximal and mid small intestine and 3-4% was found in the distal part of the small intestine. There were no significant differences in the overall distribution of orally administered radioactivity between two age groups. Based on liquid chromatography and trichloroacetic acid precipitation, a substantial amount of EGF recovered from the luminal contents (63-86%) and the gastrointestinal wall (42-81%) remained "intact". The receptor binding ability of the EGF recovered from the gastric contents was 96-102% comparable to the native EGF tracer. The receptor binding ability remained high (40-58%) in the proximal small intestinal lumen and it decreased to 15% in the distal small intestinal lumen in newborn pigs. In 5-day-old pigs, EGF recovered from the small intestinal contents had 5 to 24% receptor binding ability when compared with native EGF tracer. The receptor binding ability of the EGF recovered from all other organs was below 5% with an exception of the gastric wall, from which recovered EGF retained 9 to 26% receptor binding ability. These results indicate that most of orally ingested EGF remained in the gastrointestinal tract in neonatal pigs 45 min after oral ingestion, and significant amount of the ingested EGF remained biologically active. It suggests that milk-borne EGF can survive in the gastrointestinal tract and may play a role in regulating gut development in neonatal animals. PMID- 9734701 TI - Antioxidant status and alpha1-antiproteinase activity in subarachnoid hemorrhage patients. AB - The antiproteasic activity of alpha1-antitrypsin (alpha1-AT) is reduced in cases of subarachnoid hemorrhage from ruptured intracranial aneurysm and particularly in patients currently smoking; alpha1-AT is very sensitive to oxidant agents. About 50% of physiological anti-oxidant systemic capacity is represented by Vitamin A, E and C. Plasmatic amounts of alpha1-AT, alpha1-AT Collagenase Inhibitory Capacity (CIC) and levels of vitamin A, vitamin E and vitamin C were analyzed in 39 patients, 26 women and 13 men, operated for intracranial aneurysm; 11 patients with unruptured intracranial aneurysm were considered as controls while 28 patients were included within 12 hours from subarachnoid hemorrhage (SAH). Plasmatic levels of vitamin A and vitamin E were significantly lower (p=0.038 and p=0.0158) in patients suffering SAH than in controls, while no statistically significant differences were found in mean plasmatic vitamin C levels. Level of alpha1-AT was not statistically different in controls and in patients with SAH; however, the activity of alpha1-AT, evaluated as CIC, is significantly reduced in patients with SAH (p=0.019). We have observed that systemic plasmatic levels of vitamins did not significantly differ in relation to smoking habit. Vitamin A and E represent an important defensive system against free radicals reactions. Particularly, vitamin E acts as an antioxidant by scavenging free-radicals. A reduced anti-oxidant status might be related to the higher sensibility of alpha1-AT to oxidative reactions and the activity of alpha1 AT is dependent on the antioxidant capacity of liposoluble vitamins. We can speculate that an acute systemic oxidative stress condition might influence the rupture of intracranial aneurysms. PMID- 9734702 TI - Enhancement of peroxynitrite-evoked acetylcholine release by hydroxyl radical scavengers from mouse cerebral cortical neurons. AB - We investigated the effects of hydroxyl radical scavengers on peroxynitrite (OONO )-evoked acetylcholine (ACh) release from mouse cerebral cortical neurons. N,N' dimethylthiourea, a hydroxyl radical scavenger, dose-dependently increased OONO( )-evoked ACh release. Other hydroxyl radical scavengers such as uric acid and mannitol, also enhanced OONO(-)-evoked ACh release, although these enhancing effects were not found in the absence of OONO-. In addition, OONO(-)-induced [45Ca2+]influx was significantly facilitated by the scavengers, whereas no effects of the scavengers on [45Ca2+]influx was observed in the absence of OONO-. These results indicate that hydroxyl radical scavengers enhance OONO(-)-evoked ACh release via the facilitation of OONO(-)-induced [45Ca2+]influx. PMID- 9734703 TI - Lack of calmodulin antagonism of melatonin in T-lymphocyte activation. AB - Despite various reported effects of the pineal hormone melatonin on the immune system, its mechanism of action on immune cells is still unknown. Since melatonin has been suggested as a physiological antagonist to calmodulin in certain cell types, we investigated effects of melatonin on calmodulin-dependent IL-2 production and proliferation of activated T-lymphocytes. It was found, however, that, in contrast to the calmodulin antagonists trifluoperazine and W7, melatonin neither inhibited the IL-2 production of activated lymphoblastoid Jurkat T-cells nor decreased the mitogen response of peripheral blood mononuclear leukocytes. Preincubation of Jurkat cells with melatonin did not influence trifluoperazine effects on IL-2 production indicating that melatonin does not bind to the same sites of calmodulin as trifluoperazine, as has been postulated. In conclusion, these results did not give any evidence for a calmodulin antagonism of melatonin in T-lymphocyte activation. Thus, melatonin as a calmodulin antagonist appears not to be a universal phenomenon. PMID- 9734704 TI - Perinatal delta9-tetrahydrocannabinol exposure reduces proenkephalin gene expression in the caudate-putamen of adult female rats. AB - Perinatal delta9-tetrahydrocannabinol (delta9-THC) exposure in rats affects several behavioral responses, such as opiate self-administration behavior or pain sensitivity, that can be directly related to changes in opioidergic neurotransmission. In addition, we have recently reported that the administration of naloxone to animals perinatally exposed to delta9-THC produced withdrawal responses, that resemble those observed in opiate-dependent rats. The purpose of the present study was to examine the basal opioid activity in the brain of adult male and female rats that had been perinatally exposed to delta9-THC. To this aim, proenkephalin mRNA levels were measured, by using in situ hybridization histochemistry, in the caudate-putamen, nucleus accumbens, central amygdala and prefrontal cingulate cortex. The results showed a marked reduction in proenkephalin mRNA levels in the caudate-putamen of delta9-THC-exposed females as compared to oil-exposed females, whereas no changes were observed between delta9 THC- and oil-exposed males. There were no differences in proenkephalin mRNA levels in the nucleus accumbens, central amygdala and prefrontal cingulate cortex between males and females perinatally exposed to delta9-THC and their respective controls, although a certain trend to decrease was observed in delta9-THC-exposed females. In summary, perinatal exposure to delta9-THC exposure decreased proenkephalin gene expression in the caudate-putamen of adult rats, although this effect exhibited a marked sexual dimorphism since it was only seen in females. This result is in agreement with a previous observation from our laboratory that females, but not males, that had been perinatally exposed to delta9-THC, self administered more morphine in adulthood. This suggests that low levels of proenkephalin mRNA may be used as a predictor of greater vulnerability to opiates. PMID- 9734705 TI - Recording heart rate and blood pressure in rats during parabolic flight. AB - The way in which the cardiovascular system adapts to weightlessness is still under discussion. No data are yet available on the responses of rats during space flight, although this animal is commonly used in simulation studies. We have designed and tested a protocol to study the short term responses of the cardiovascular system to weightlessness during parabolic flight. A telemetry system was used to measure heart rate (HR) and blood pressure. It was possible to collect and record radio-signals without any interference. Microgravity caused a reduction in HR, an increase in mean arterial pressure (MAP, 7%), and a non significant decrease in central venous pressure (CVP, 13%). The change in CVP was similar to the decrease observed in human space flight. This type of study may also be feasible for longer exposure of rats to microgravity (space flight). PMID- 9734707 TI - Improvement by several antioxidants of macrophage function in vitro. AB - The toxic effects of oxygen radicals produced by immune cells can be controlled to certain degree by endogenous antioxidants, because of their scavenger action. This control is specially important in a type of immune cell, i.e.: the phagocyte, which needs oxygen free radicals and uses antioxidants in order to support its functions. Previous studies have shown an stimulation of the immune system with an antioxidant enriched diet. In the present work, we have studied the effects in vitro of several antioxidants: alpha-tocopherol or vitamin E (VE), ascorbic acid (AA), glutathione (GSH), N-acetylcysteine (NAC) and thioproline or thiazolidine-4-carboxylic acid (TCA), at different concentrations, on the various steps of the phagocytic process of murine peritoneal macrophages, i.e.: adherence to substrate, migration (random migration and directed migration or chemotaxis), ingestion and superoxide anion production. The results show an antioxidant induced stimulation of the phagocytic process of macrophages. Thus, the adherence to substrate was raised, after short incubation times, by a-tocopherol and ascorbic acid. Random migration, chemotaxis, ingestion and superoxide anion production were increased by all the antioxidants used. PMID- 9734706 TI - Adrenergic-mediated effects of cocaine on the myocardial force-frequency relationship. AB - We studied the role of sarcolemmal alpha- and beta-adrenoceptors activation in the effects of cocaine on the positive force staircase in isolated guinea pig atria. The preparations were superfused with Tyrode's solution at 31 degrees C while attached to a force transducer to measure peak tension developed (PTD), maximum velocity of development of tension (Vmax T) and time to peak tension (TPT). The positive force staircase was not affected by propranolol or phentolamine, but it was abolished by nifedipine. Cocaine 1 mg/l (2.9 microM) enhanced PTD and Vmax T, while TPT remained unchanged. On the other hand, cocaine did not modify the increase in PTD induced by the increase in frequency of stimulation, but significantly reduced the magnitude of the increase in Vmax T. The cocaine-induced attenuation of the increase in Vmax T in response to changes in the frequency of stimulation was abolished by both propranolol and phentolamine. It is concluded that the effect of cocaine on the force-frequency relationship required background activation of alpha- and beta-adrenergic receptors. PMID- 9734708 TI - Hair analysis for drugs of abuse XX. Incorporation and behaviors of seven methamphetamine homologs in the rat hair root. AB - To elucidate drug disposition in hair, the incorporation and retention behavior of 7 phenethylamines in the rat hair root were investigated: methamphetamine(MA), 3,4-methylenedioxymethamphetamine(MDMA), benzphetamine(BZP), ephedrine(EP), N,N dimethylamphetamine(DMA), p-nitro-methamphetamine(NO2MA), and N acetylmethamphetamine(AcMA). On day 10 after shaving the hair on the back of the rats, drug was intraperitoneally administered at a single dose of 10 mg/kg to Long Evans rats (which were male and 6 weeks of age), possessing black and white hair, and the back hair that grew was collected by plucking with hair nippers at 0.083 h (5 min), 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 9 h, 24 h, 33 h and 48 h. After washing the plucked hairs three times with 0.1% sodium dodecylsulfate, the amount of drug in each of the hair root samples was analyzed by a selected ion monitoring of gas chromatography mass spectrometry (GC-MS) analysis. The times at which the concentration of each drug in the hair root samples reached the peak concentration, ranged between 3.30 and 41.51 ng/mg. For each drug, the point of time at which the largest positive incremental change in drug concentration was seen, ranged between 5 min and 1 h, for all of the drugs except for AcMA which was hardly incorporated in the rat hair. The data showed that there are mainly 4 modes in which a drug becomes incorporated into the black hair root: rapid and prolonged incorporation (NO2MA, MDMA), rapid and short incorporation(MA, DMA), slow and prolonged incorporation(BZP, EP), slow and short incorporation, which includes hardly any incorporation (AcMA). As all seven drugs were hardly incorporated into the white hair, it was concluded that the combination of melanin and basic compounds is essential for a drug to become incorporated into hair. Our results suggest that a portion of the drugs in the hair root is accumulated in the hair shaft, and the remaining portion is redistributed outside the hair shaft. The second finding is that the concentration of drug incorporated into hair mainly depends on two processes--(1) the drug incorporation into hair and the drug retention in hair. PMID- 9734709 TI - The phospholipase C inhibitor U73122 increases cytosolic calcium in MDCK cells by activating calcium influx and releasing stored calcium. AB - The effects of the phospholipase C (PLC) inhibitor U73122 on intracellular calcium levels ([Ca2+]i) were studied in MDCK cells. U73122 elevated [Ca2+]i dose dependently. Ca2+ influx contributed to 75% of 20 microM U73122-induced Ca2+ signals. U73122 pretreatment abolished the [Ca2+]i transients evoked by ATP and bradykinin, suggesting that U73122 inhibited PLC. The Ca2+ signals among individual cells varied considerably. The internal Ca2+ source for the U73122 response was the endoplasmic reticulum (ER) since the response was abolished by thapsigargin. The depletion of the ER Ca2+ store triggered a La3+-sensitive capacitative Ca2+ entry. Independently of the internal release and capacitative Ca2 entry, U73122 directly evoked Ca2+ influx through a La3+-insensitive pathway. The U73122 response was augmented by pretreatment of carbonylcyanide m chlorophynylhydrozone (CCCP), but not by Na+ removal, implicating that mitochondria contributed significantly in buffering the Ca2+ signal, and that efflux via Na+/Ca2+ exchange was insignificant. PMID- 9734710 TI - Endothelin converting enzyme inhibitor protects development of right ventricular overload and medial thickening of pulmonary arteries in rats with monocrotaline induced pulmonary hypertension. AB - We evaluated the effects of FR901533, endothelin converting enzyme inhibitor, on development of right ventricular overload and medial thickening of pulmonary arteries in rats with monocrotaline-induced pulmonary hypertension. Pulmonary hypertension was induced by a single injection of monocrotaline (80 mg/kg). Twenty-four hours later (day 1), continuous subcutaneous injection of FR901533 (100 mg/kg/day) was started. Right ventricular systolic pressure, mass ratio of right ventricle to left ventricle, right ventricular wall thickness, right ventricular myocardial fiber diameter, percent medial thickness, and percent smooth muscle area in pulmonary arteries were significantly less in rats that received FR901533 than in the control with monocrotaline on day 28. Both immunoreactivities of endothelin-1 in pulmonary arteries and plasma endothelin-1 levels were observed significantly less in rats treated with FR901533 than in the control with monocrotaline. There were significant increased immunoreactivities of endothelin-B receptor in pulmonary arteries in rats that received FR901533 as compared with those in the control with monocrotaline. FR901533 (100 mg/kg/day), protected the development of right ventricular overload and medial thickening of pulmonary arteries in a rat model of pulmonary hypertension. PMID- 9734711 TI - Contribution of caffeine and flavanols in the induction of hepatic Phase II activities by green tea. AB - Aqueous extracts of green tea, at concentrations of 2.5. 5.0 and 7.5%, were administered to rats as the sole drinking fluid for 4 weeks. Hepatic glutathione S-transferase (GST) activity, determined using 1-chloro-2,4-dinitrobenzene (CDNB) and 3,4-dichloronitrobenzene (DCNB) as substrates, and UDP-glucuronosyl transferase activity, determined using 2-aminophenol as substrate, were induced but the effect was not always dose dependent. At the two highest doses, hepatic catalase activity was inhibited. In a second study, animals were exposed for 4 weeks to aqueous extracts (2.5%, v/v) of green tea, black tea (which has a much lower content of flavanols compared with green tea) and decaffeinated black tea. Treatment with the black tea enhanced GST activity, whether monitored using CDNB or DCNB, and the glucuronidation of 2-aminophenol. Treatment with decaffeinated black tea failed to modulate any of these activities, whereas treatment with green tea only enhanced the glucuronidation of 2-aminophenol. Finally, at this concentration of tea extract administration, black and decaffeinated black tea, but not green tea, suppressed catalase activity. It is concluded that neither flavanols nor caffeine are responsible for the induction of hepatic Phase II activities and inhibition of catalase activity in the rat. PMID- 9734712 TI - Characterization of flavonoids as monofunctional or bifunctional inducers of quinone reductase in murine hepatoma cell lines. AB - The ability of flavonoid compounds to induce the activity of the phase II anticarcinogenic marker enzyme, quinone reductase (QR), has been studied in a wild-type murine hepatoma cell line (Hepalclc7) and in an Ah-receptor-defective mutant of the same cell line (Hepalclc7 bp(r)cl). The results showed that 10 (beta-naphthoflavone, kaempferide, tamarixetin, rhamnetin, quercetin, kaempferol, quercetin-4'-glucoside, isorhamnetin, daidzein and genistein) of the 13 flavonoids tested induced QR activity in the wild-type cells. Only the latter six also showed such activity in the bp(r)cl mutant, which indicates that they induce phase II enzymes directly (monofunctional inducers), whereas the others induce phase 11 enzymes only in cells with an operative Ah receptor system (bifunctional inducers). The metabolism of representatives of monofunctional (quercetin) and bifunctional (tamarixetin and rhamnetin) flavonol inducers were studied in both wild-type and bp(r)cl cells. In all cases, the major metabolites were glucuronides. Quercetin produced identical metabolites in both cell types, whereas one glucuronide of tamarixetin and two glucuronides of rhamnetin were not formed in the mutant cells. This shows that flavonoids can be mono- or bifunctional inducers depending on their chemical structure, and that the glucuronidation pattern of bifunctional inducers is altered by the presence of a functional Ah receptor system. PMID- 9734713 TI - Promotional effect of N-nitroso-N-(3-keto-1,2-butanediol)-3'-nitrotyramine (a nitrosated Maillard reaction product) in mouse fibroblast cells. AB - N-Nitroso-N-(3-keto-1,2-butanediol)-3'-nitrotyramine (NO-NTA) is a product of model browning system generated in the presence of sodium nitrite. The chemical structure of this compound has been confirmed by UV, mass and nuclear magnetic resonance, and infrared spectroscopy in our previous study. A two-stage transformation protocol was used to chemically transform the mouse embryo fibroblasts C3H10T1/2 cells. To initiate transformation, the cells were treated with benzo[a]pyrene (BaP) (0.1 mg/ml), and NO-NTA (0.01, 0.1 and 1 mg/ml) was employed subsequently to complete the transformation process. Malignant transformed foci were formed in BaP-initiated and NO-NTA promoted C3H10T1/2 cells after 8 wk. Cells treated with NO-NTA alone failed to induce transformation. However, cells initiated with BaP and promoted by cells initiated with BaP and promoted by NO-NTA demonstrated oncogenic properties. Cell lines transformed with NO-NTA-transformed colonies exhibited enhanced growth rate, anchorage independence and tumorigenicity in animals relative to parent cells. These results indicate that NO-NTA is a new tumour promoter and may induce tumour promotion by two-stage oncogenesis. Further studies on the mechanism of action of NO-NTA are now in progress. PMID- 9734714 TI - Protective effects of fruits and vegetables against in vivo clastogenicity of cyclosphosphamide or benzo[a]pyrene in mice. AB - Seven fruits and 10 vegetables commonly consumed in Germany were investigated for their anticlastogenic potencies against cyclophosphamide (CP) and benzo[a]pyrene (BaP) in the in vivo mouse bone marrow micronucleus assay. We detected protective effects in 76.5% and 70.6% of the samples, respectively, and more or less distinct quantitative differences between the various plant materials and the two clastogens investigated. With respect to CP, moderate activities were exerted by sweet cherries, strawberries, cucumber, radish and tomatoes, average activities by bananas, oranges, peaches, asparagus and red beets and strong activities by yellow red peppers and especially spinach. Apples (cultivar Jona Gold), brussels sprouts, cauliflower and onions were inactive. With respect to BaP, we found moderate activities in strawberries, brussels sprouts and radish, average activities in sweet cherries, oranges, peaches, asparagus, red beets, cucumber and spinach and strong activities in bananas and kiwi. Apples, cauliflower, onions, tomatoes and yellow-red peppers were inactive. When oranges were fractionated according to previously described schemes (Edenharder et al., 1995), anticlastogenic activities against CP were exerted by materials extracted with n hexane, acetone and 2-propanol and in the terminal residue, but not in the dichloromethane and water phases. With respect to BaP, materials extracted with acetone showed strong anticlastogenicity while the 2-propanol fraction, the aqueous phase and the terminal residue were less potent. The n-hexane and the dichloromethane fractions were inactive. In red beets, all fractions showed anticlastogenicity against CP and BaP as well. However, the n-hexane and dichloromethane fractions were most potent with respect to CP, while for BaP the aqueous phase and the terminal residue were most effective. These result suggest the presence of various (groups of) anticlastogenic compounds with different chemical structure. PMID- 9734715 TI - Lack of effect of coumarin on unscheduled DNA synthesis in precision-cut human liver slices. AB - In this study the effect of coumarin on unscheduled DNA synthesis (UDS) in precision-cut human liver slices has been examined. Liver slices from tissue samples from four donors were cultured for 24 hr in medium containing [3H]thymidine and 0-5.0 mM coumarin using a dynamic organ culture system and processed for autoradiographic evaluation of UDS. As positive controls liver slices were also cultured with three known genotoxic agents, namely 0.02 and 0.05 mM 2-acetylaminofluorene (2-AAF), 0.002 and 0.02 mM aflatoxin B1 (AFB1) and 0.005 and 0.05 mM 2-amino-1-methyl-6-phenylimidazo [4,5-b]pyridine (PhIP). UDS was quantified as the net grain count in centrilobular hepatocytes and as the percentage of centrilobular hepatocyte nuclei with more than five net grains. Compared with control liver slice cultures, treatment with 0.05-5.0 mM coumarin had no effect on UDS. In contrast, treatment with 0.02 and 0.05 mM 2-AAF, 0.002 and 0.02 mM AFB1 and 0.005 and 0.05 mM PhIP produced significant increases in the net grain counts of centrilobular hepatocytes. The greatest induction of UDS was observed in liver slices treated with 0.05 mM PhIP. Treatment with 2-AAF, AFB1 and PhIP also produced significant increases in the number of centrilobular hepatocyte nuclei with more than five net grains. At the concentrations examined neither coumarin. 2-AAF, AFB1 nor PhIP had any significant effect on replicative DNA synthesis in 24 hr cultured human liver slices. These results demonstrate that coumarin does not induce UDS in cultured human liver slices. However, all three positive control compounds produced marked significant increases in UDS, thus confirming the functional viability of the human liver slice preparations used in this study. The results of this study suggest that coumarin is not a genotoxic agent in human liver. PMID- 9734716 TI - Effects of singly administered betaine on hepatotoxicity of chloroform in mice. AB - Effects of a single dose of betaine on the chloroform-induced hepatotoxicity were examined in adult male ICR mice. Administration of betaine (1000 mg/kg, ip) 1 to 7 hr prior to a chloroform challenge (0.25 ml/kg, ip) resulted in remarkable enhancement of hepatotoxicity as indicated by increases in serum sorbitol dehydrogenase (SDH), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities. The potentiation of hepatotoxicity was most significant when mice were treated with betaine 4 hr earlier than chloroform. However, a 24 hr prior administration of betaine protected the animals from induction of the chloroform hepatotoxicity. Thus, its effect appeared to be highly dependent on the time lapse from the betaine pretreatment to the challenge of mice with chloroform. Betaine treated either 4 or 24 hr prior to sacrifice did not alter the hepatic contents of cytochrome P-450, cytochrome b5, or NADPH cytochrome P-450 reductase activity. Accordingly the hepatic microsomal p nitroanisole O-demethylase, aminopyrine N-demethylase, or p-nitrophenol hydroxylase activities were not influenced by the betaine pretreatment. Betaine was shown not to affect any of the enzyme activities associated with glutathione (GSH) conjugation reaction, such as glutathione S-transferases (GSTs), glutathione disulfide (GSSG) reductase and GSH peroxidase irrespective of the time of its administration. When betaine was administered to mice 2-6 hr prior to sacrifice, hepatic GSH level, but not plasma GSH, was decreased significantly. Enhancement of the chloroform hepatotoxicity by betaine correlated well with the reduction in hepatic GSH levels. Both hepatic and plasma GSH levels were elevated in mice 24 hr following the betaine treatment. The results suggest that betaine affects induction of the chloroform hepatotoxicity by modulating the availability of hepatic GSH, which appears to be associated with its role in the transsulfuration pathway in the liver. PMID- 9734717 TI - The association of increasing dietary concentrations of fish oil with hepatotoxic effects and a higher degree of aorta atherosclerosis in the ad lib.-fed rabbit. AB - The long-term effects of consumption of marine long-chain n-3 polyunsaturated fatty acids (PUFA) on atherosclerosis in the rabbit were examined. Female Dutch rabbits were fed purified diets, containing 40 energy% total fat, for a period of 2.5 years. To study the dose response relationship between fish oil intake and atherosclerosis, four diets were formulated with fish oil levels being 0, 1, 10 and 20 energy%. A fifth and sixth group were fed an alpha-linolenic acid-(C18:3, n-3) and linoleic acid-(C18:2, n-6) rich diet, respectively. Every 6 weeks, blood samples were taken for determination of clinical chemical parameters, triacylglycerol and total cholesterol levels. Feeding 10 and 20 energy% fish oil containing diets, resulted in an increase of liver enzymes (AST, ALT and ALP). Histological evaluation of the liver also revealed adverse effects of fish oil containing diets. Triacylglycerol blood levels were similar in all groups, and remained constant throughout the study. Total cholesterol levels in blood was significantly lower in the animals fed a linoleic acid-rich diet, as compared with the other five groups. An n-3 long-chain PUFA concentration dependent increase in aorta plaque surface area was observed in the fish oil groups. A significant positive relationship was found between the group mean score for severity of liver pathology and the aorta plaque surface area. These results indicate that the long-chain n-3 polyunsaturated fatty acids in fish oil may be hepatotoxic to the herbivorous rabbit, which may interfere with the outcome of atherosclerosis studies. This finding necessitates the exclusion of liver pathology in experimental studies on atherosclerosis in animal models. PMID- 9734718 TI - Effects of fumonisin B1 in pregnant rats. Part 2. AB - The developmental toxicity of purified fumonisin B1 (FB1), a mycotoxin from the common corn fungus Fusarium moniliforme, was examined in Charles River rats. Pregnant rats were dosed orally on gestation days 3-16 at 0, 6.25, 12.5, 25 or 50 mg FB1/kg body weight/day. FB1 was not teratogenic at the doses tested. At 50 mg/kg, maternal toxicity (inappetence, emaciation, lethargy, death, resorption of entire litters) and foetal toxicity (increased number of late deaths, decreased foetal body weight, decreased crown rump length, increased incidence of hydrocephalus, increased incidence of skeletal anomalies) were seen. The foetal toxicity observed at 50 mg/kg may be related to maternal toxicity. Histopathological evaluation of tissues from dams of control and all treated groups revealed dose-related toxic changes in kidney and liver tissues. Acute toxic tubular nephrosis was seen in kidneys from all treated groups. Hepatocellular cytoplasmic alteration and individual cellular necrosis of the liver was seen in the two high-dose groups. Sphinganine (Sa) and sphingosine (So) were measured in day-17 adult and foetal tissues. Dose related increases in Sa/So ratios were seen in maternal liver, kidney, serum and brain, but there was no effect on foetal liver, kidney and brain. These data suggest that FB1 does not cross the placenta and further suggest that the observed foetal toxicity is a secondary response to maternal toxicity. PMID- 9734719 TI - Studies on the potential for genotoxic carcinogenicity of fragrances and other chemicals. AB - The potential of fragrances, physiological chemicals, natural products and a group of randomly selected chemicals to induce cancers by a genotoxic mechanism (i.e. "genotoxic" carcinogenesis) was compared using structure-activity relationships (SAR) models. Fragrances are significantly less likely to induce genotoxic carcinogenicity than randomly selected chemicals or natural products. With respect to the latter potential, fragrances were indistinguishable from normal mammalian physiological constituents. PMID- 9734720 TI - Induction of thyroid tumours in (C57BL/6N x C3H/N)F1 mice by oral administration of kojic acid. AB - The tumorigenicity of kojic acid (KA), which is widely used for food and cosmetics in Japan, was examined in B6C3F1 mice. Female and male animals were divided into three groups and given 0, 1.5 and 3.0% KA containing food from the age of 6 weeks. At sacrifice after 20 months, thyroid weights were significantly increased in both sexes of mice receiving KA, especially in the male groups. The enlarged thyroid glands histologically featured diffuse hyperplasia and follicular adenomas, the incidences of the latter being 65% and 87%, respectively in 1.5% and 3.0% KA-treated males, significantly higher than the control value of 2%. In the females, the figures were 2%, 8% and 80% in the 0%, 1.5% and 3.0% KA groups, respectively. The serum free T3 levels in the 3.0% KA animals of both sexes at month 6 were significantly lower than in the controls. On the other hand, their serum TSH levels were higher, although the differences disappeared at later time points. In conclusion, continuous administration of high dose of KA induces thyroid adenomas in male and female B6C3F, mice, presumably by a mechanism involving decrease in serum free T3 levels and increased TSH. PMID- 9734721 TI - Subchronic oral toxicity study of Aquacoat ECD ethylcellulose aqueous dispersion in the rat. AB - Groups of 20 male and 20 female Sprague-Dawley rats were administered undiluted Aquacoat ECD ethylcellulose aqueous dispersion by oral gavage at doses of 903, 2709 or 4515 mg/kg body weight/day (dry weight basis) for 90 days. Control animals received water at the same dosage volume as the high-dose group. Body weights and food consumption were recorded weekly. Blood was collected prior to study termination for haematology and clinical chemistry measurements. Survivors underwent complete necropsies on days 91 94. Selected organs were weighed and histologically examined. The only treatment-related clinical sign observed was pale faeces which was noted among males and females receiving 2709 and 4515 mg/kg/day Aquacoat ECD. No statistically significant differences in body weights, body weight gains, food consumption and organ weights were noted among males and females when compared with controls. No treatment-related effects in haematology parameters were noted. Significantly decreased total protein and globulin levels and increases in alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in male rats receiving 2709 and 4515 mg Aquacoat ECD/kg/day were considered to be treatment related. No gross or microscopic lesions were attributed to Aquacoat ECD treatment. Under the conditions of this study, the no-observed adverse-effect level (NOAEL) for female rats is in excess of 4515 mg/kg/day: the NOAEL for male rats is 903 mg/kg/day. PMID- 9734723 TI - HIV infection: a model chronic illness for studying wasting diseases. PMID- 9734722 TI - Mineral intake and blood pressure in African Americans. PMID- 9734724 TI - Further evidence of the cardiovascular benefits of diets enriched in carotenoids. PMID- 9734725 TI - Nutrition and catch-up bone augmentation in young women. PMID- 9734726 TI - Glucagon-like peptide 1 increases the period of postprandial satiety and slows gastric emptying in obese men. AB - The gut peptide glucagon-like peptide 1(7-36) amide (GLP-1) is released into the circulation after food intake. GLP-1 has been shown to have an incretin effect and inhibits gastrointestinal motility in humans. In rats, intracerebral administration of GLP-1 results in reduced food intake. Obese humans have been found to have an attenuated plasma GLP-1 response to a mixed meal. To approximate the physiologic state, GLP-1 or saline was administered intravenously and randomly at the beginning of a test meal served on a universal eating monitor to 6 obese subjects to test our hypothesis that GLP-1 influences termination of food intake (and thus food intake during a meal) and feelings of satiety in humans. As a marker for gastric emptying, 1.5 g acetaminophen was given at the start of the meal. Blood samples for analysis of acetaminophen, insulin, glucose, glucagon, and C-peptide were obtained. Hunger, fullness, and food choice were assessed with visual analogue scales and food-choice questionnaires. GLP-1 infusion resulted in a prolonged period of reduced feelings of hunger, desire to eat, and prospective consumption after the meal. The rate of gastric emptying was slower during infusion of GLP-1. Postprandial blood glucose concentrations were reduced during the GLP-1 infusion, but the amount of energy consumed, eating rate, and plasma concentrations of insulin, glucagon, and C-peptide were unchanged. GLP-1 given exogenously at the start of a meal did not seem to affect meal termination or the amount of food eaten. However, postprandial feelings of hunger decreased, suggesting that exogenous GLP-1 may influence feelings of hunger and satiety in humans. PMID- 9734727 TI - Aspartame: neuropsychologic and neurophysiologic evaluation of acute and chronic effects. AB - BACKGROUND: Neurobehavioral symptoms have been reported anecdotally with aspartame. OBJECTIVE: This study sought to determine whether aspartame can disrupt cognitive, neurophysiologic, or behavioral functioning in normal individuals. DESIGN: Forty-eight healthy volunteers completed a randomized, double-blind, placebo-controlled, crossover study. The first month was aspartame free. Subjects then consumed sodas and capsules with placebo, aspartame, or sucrose for 20 d each. Order was randomized and subjects were assigned to either a high- (45 mg x kg body wt(-1) x d(-1)) or low- (15 mg x kg body wt(-1) x d(-1)) dose aspartame group. Neuropsychologic and laboratory testing was done on day 10 of each treatment period to determine possible acute effects and on day 20 for possible chronic effects. RESULTS: Plasma phenylalanine concentrations increased significantly during aspartame treatment. Neuropsychologic results; adverse experiences; amino acid, insulin, and glucose values; and electroencephalograms were compared by sex and by treatment. No significant differences were found for any dependent measure. CONCLUSION: Large daily doses of aspartame had no effect on neuropsychologic, neurophysiologic, or behavioral functioning in healthy young adults. PMID- 9734728 TI - Enrichment of hen eggs with n-3 long-chain fatty acids and evaluation of enriched eggs in humans. AB - Eggs enriched with n-3 polyunsaturated fatty acids (PUFAs) were produced by hens fed diets containing fish oil or a combination of fish and vegetable oils. In a sensory evaluation, 78 untrained volunteers could not distinguish between ordinary and enriched eggs. Storage life was also not significantly different between egg types. A food intake survey of 4 groups of 14 subjects each who consumed 7 eggs/wk for 24 wk showed that intakes of the major dietary components were not significantly different for 4 different egg types. Mean (n = 56) plasma cholesterol and triacylglycerol concentrations were not significantly different at the start and finish of the study. Body weight and HDL concentrations increased during the study (P < 0.05). For the last 2 wk of the experiment (weeks 23-24), mean egg consumption was increased from a total of 14 to a total of 21 eggs, resulting in a small increase in plasma triacylglycerols only. There were no significant differences (P > 0.05) in body weight, blood pressure, or plasma lipid components among treatment groups consuming the 4 different egg types. Blood samples taken after 16 and 22 wk from fasted subjects showed significant increases in eicosapentaenoic acid, docosahexaenoic acid, and total n-3 PUFAs in subjects consuming enriched eggs compared with controls. In addition, the ratio of n-6 to n-3 PUFAs in plasma was significantly reduced from 12.2:1 to 6.5-7.7:1 in subjects consuming enriched eggs compared with controls. Consumption of only one enriched egg daily can contribute substantially to the recommended daily intake of n-3 PUFAs. PMID- 9734729 TI - Long-term intake of soy protein improves blood lipid profiles and increases mononuclear cell low-density-lipoprotein receptor messenger RNA in hypercholesterolemic, postmenopausal women. AB - The long-term clinical effects of soy protein containing various amounts of isoflavones on lipoproteins, mononuclear cell LDL receptor messenger RNA concentrations, and other selected cardiovascular risk factors are not well known. Sixty-six hypercholesterolemic, free-living, postmenopausal women were investigated during a 6-mo parallel-group, double-blind trial with 3 interventions. After a control period of 14 d, all subjects were randomly assigned to 1 of 3 dietary groups (all with 40 g protein): a National Cholesterol Education Program (NCEP) Step 1 diet with protein from casein and nonfat dry milk (control), an NCEP Step 1 diet with protein from isolated soy protein containing moderate amounts of isoflavones (ISP56), or an NCEP Step 1 diet with protein from isolated soy protein containing high amounts of isoflavones (ISP90). Non-HDL cholesterol in both the ISP56 and ISP90 groups was reduced compared with the control group (P < 0.05), whereas total cholesterol was not changed. HDL cholesterol increased in both the ISP56 and ISP90 groups (P < 0.05), whereas the ratio of total to HDL cholesterol decreased significantly in both groups compared with the control (P < 0.05). Mononuclear cell LDL receptor messenger RNA concentrations increased in subjects consuming ISP56 or ISP90 compared with the control (P < 0.05). These results indicate that soy protein, with different amounts of isoflavones, may decrease the risk of cardiovascular disease via improved blood lipid profiles, and that the mechanism by which apolipoprotein B containing lipoproteins were depressed may be via alterations in LDL receptor quantity or activity. PMID- 9734731 TI - Lipoprotein secretion by intestinal Caco-2 cells is affected differently by trans and cis unsaturated fatty acids: effect of carbon chain length and position of the double bond. AB - The effects of trans fatty acids on intestinal lipoprotein secretion were determined in polarized Caco-2 cells. Palmitic acid (16:0), palmitoleic acid (c 16:1delta9), and palmitelaidic acid (t-16:1delta9), as well as stearic acid (18:0), oleic acid (c-18:1delta9), c-vaccenic acid (c-18:1delta11), elaidic acid (t-18:1delta9), and t-vaccenic acid (t-18:1delta11) were studied. Compared with 18:0 (control), c- and t-18:1delta9 increased triacylglycerol secretion (2.7- and 3.6-fold, respectively) as well as apolipoprotein (apo) B-48 and apo B-100 secretion (both 1.6-fold compared with 18:0); c- and t-18:1delta11 caused a modest 1.7-fold increase in triacylglycerol secretion with no significant effect on secretion of apo B. Thus, the position of the double bond in the 18:1 isomers, but not its geometrical configuration, affected lipoprotein secretion by Caco-2 cells. In contrast, the effects of the geometrical isomers (cis and trans) of C16 fatty acids were not comparable: t-16:1delta9 did not affect triacylglycerol and apo B secretion (compared with 16:0, as control) whereas c-16:1delta9 was a potent stimulator of secretion of triacylglycerol (2.4-fold higher than 16:0), apo B-48 (1.3-fold higher than 16:0), and apo B-100 (1.5-fold higher than 16:0). We conclude that the carbon chain length of fatty acids, as well as the position of double bonds and their stereochemical configuration, are important determinants of the unique effects of various species of dietary trans fatty acids on lipoprotein secretion and composition in Caco-2 cells. PMID- 9734730 TI - Effect of long-term olive oil dietary intervention on postprandial triacylglycerol and factor VII metabolism. AB - Although the beneficial effects of Mediterranean-type diets, which are rich in olive oil, a good source of monounsaturated fatty acids (MUFAs), are generally accepted, little is known about the effects of long-term dietary MUFA intake on postprandial lipoprotein metabolism and hemostasis. This study used a single blind, randomized, crossover design to investigate the relative effects of a long term dietary olive oil intervention and a control [saturated fatty acid (SFA) enriched] diet on postprandial triacylglycerol metabolism and factor VII activity. The postprandial response to a standard test meal was investigated in 23 healthy men who adhered to both diets for 8 wk. cis-MUFAs were successfully substituted for SFAs in the MUFA diet without affecting total dietary fat or energy intakes. The long-term dietary MUFA intervention significantly reduced plasma and LDL-cholesterol concentrations (P = 0.01). Postprandial triacylglycerol concentrations were significantly greater in the early postprandial period after the MUFA diet (P = 0.003). Postprandial factor VII activation and the concentration of the factor VII antigen were significantly lower after the MUFA diet (P = 0.04 and P = 0.006, respectively). This study showed that isoenergetic substitution of MUFAs for SFAs reduces plasma cholesterol and reduces the degree of postprandial factor VII activation. The alterations in the postprandial triacylglycerol response suggest a greater rate of dietary fat absorption and postprandial triacylglycerol metabolism after a diet rich in MUFAs. This study presents new insights into the biochemical basis of the beneficial effects associated with long-term dietary MUFA consumption, which may explain the lower rates of coronary mortality in Mediterranean regions. PMID- 9734732 TI - Relation between dietary fiber consumption and fibrinogen and plasminogen activator inhibitor type 1: The National Heart, Lung, and Blood Institute Family Heart Study. AB - Considerable evidence suggests that high plasma concentrations of plasminogen activator inhibitor type 1 (PAI-1) and fibrinogen increase the risk of cardiovascular disease. Recent studies report beneficial effects of dietary fiber on coronary artery disease, although the mechanisms by which high fiber intake reduces the risk of heart disease are not well understood. This study examined the relation of dietary fiber intake to PAI-1 and fibrinogen concentrations in 883 men and 1116 women aged 50.4 +/- 13.8 and 52.1 +/- 13.7 y, respectively, in the National Heart, Lung, and Blood Institute Family Heart Study. Diet was assessed with a semiquantitative food-frequency questionnaire. The natural logarithm was used to transform PAI-1 because of a skewed distribution. In the first through fifth age- and energy-specific quintiles of fiber intake, mean (ln)PAI-1 was 6.09, 5.91, 5.88, 5.82, and 5.67 pmol/L, respectively, for men and 5.50, 5.37, 5.39, 5.23, and 5.18 pmol/L, respectively, for women. Multiple regression showed that when the lowest was compared with the second, third, fourth, and fifth age- and energy-specific quintiles of fiber intake, (ln)PAI-1 was 0.21, 0.25, 0.22, and 0.32 pmol/L lower in men (P for trend = 0.009) and 0.08, 0.06, 0.14, and 0.20 pmol/L lower in women (P for trend = 0.037), respectively, with anthropometric, lifestyle, and metabolic factors adjusted for. No significant association was found between fiber intake and fibrinogen. Waist hip ratio did not modify the relation of fiber intake to PAI-1 (P for interaction = 0.39 for men and 0.36 for women). These data suggest that higher fiber intake is inversely associated with PAI-1, but not with fibrinogen concentration. PMID- 9734734 TI - High body fatness, but not low fat-free mass, predicts disability in older men and women: the Cardiovascular Health Study. AB - Using data from the Cardiovascular Health Study, we studied the relation between body composition (fat mass and fat-free mass, assessed by bioelectrical impedance) and self-reported, mobility-related disability (difficulty walking or stair climbing) in 2714 women and 2095 men aged 65-100 y. In a cross-sectional analysis at baseline (1989-1990), disability was reported by 26.5% of the women and 16.9% of the men. A positive association was observed between fat mass and disability. The odds ratio for disability in the highest quintile of fat mass was 3.04 (95% CI: 2.18, 4.25) for women and 2.77 (95% CI: 1.82, 4.23) for men compared with those in the lowest quintile. Low fat-free mass was not associated with a higher prevalence of disability. In a longitudinal analysis among persons not reporting disability at baseline, 20.3% of the women and 14.8% of the men reported disability 3 y later. Fat mass at baseline was predictive of disability 3 y later, with odds ratios of 2.83 (95% CI: 1.80, 4.46) for women and 1.72 (95% CI: 1.03, 2.85) for men in the highest quintile of fat. The increased risk was not explained by age, physical activity, chronic disease, or other potential confounders. Low fat-free mass was not predictive of disability. The results showed that high body fatness is an independent predictor of mobility-related disability in older men and women. These findings suggest that high body fatness in old age should be avoided to decrease the risk of disability. PMID- 9734733 TI - Estimation of net endogenous noncarbonic acid production in humans from diet potassium and protein contents. AB - Normal adult humans eating Western diets have chronic, low-grade metabolic acidosis, the severity of which is determined in part by the net rate of endogenous noncarbonic acid production (NEAP), which varies with diet. To prevent or reverse age-related sequelae of such diet-dependent acidosis (eg, bone and muscle loss), methods are needed for estimating and regulating NEAP. Because NEAP is difficult to measure directly, we sought a simple method to estimate it from diet-composition data. We focused on protein and potassium contents because the production of sulfuric acid from protein metabolism and bicarbonate from dietary potassium salts of organic acids are the major variable components of NEAP. Using steady state renal net acid excretion (RNAE) as an index of NEAP in 141 normal subjects eating 20 different diets, we found by multiple linear regression analysis that RNAE [mEq/d x 10460 kJ diet (mEq/d 2500 kcal)] was predictable (R2 = 0.62) from protein [g/d x 10460 kJ diet (g/d 2500 kcal); positive regression coefficient, P < 0.001] and potassium [mEq/d x 10460 kJ diet (mEq/d x 2500 kcal): negative regression coefficient, P = 0.001] contents, which were not themselves correlated. Among diets, 71% of the variation in RNAE could be accounted for by the ratio of protein (Pro) to potassium (K) content: RNAE = 62Pro/K - 17.9 (r = 0.84, R2 = 0.71, P < 0.001). Thus, by considering both the acidifying effect of protein and the alkalinizing effect of potassium (organic anions), NEAP can be predicted with confidence from the readily available contents of only 2 nutrients in foods. Provisionally, these findings allow estimation and regulation of NEAP through diet modification. PMID- 9734735 TI - Interaction of insulin, glucagon-like peptide 1, gastric inhibitory polypeptide, and appetite in response to intraduodenal carbohydrate. AB - The relation between gastrointestinal incretin hormones in the control of insulin release and short-term satiety by intestinal carbohydrate was investigated in 8 fasted, healthy male volunteers. Insulin, gastric inhibitory polypeptide (GIP), glucagon-like peptide 1 (GLP-1), and appetite ratings were measured during, and food intake was measured after, intraduodenal infusions of glucose or saline. Studies were conducted under hyperinsulinemic and euglycemic conditions. Raising plasma insulin with intravenous insulin infusion to concentrations slightly above usual postprandial concentrations (356.4 +/- 4.8 pmol/L) had no effect on GIP, GLP-1, or appetite ratings before the intraduodenal infusions began. Intraduodenal glucose infusion resulted in a further increase in plasma insulin to a peak of 779.4 +/- 114.0 pmol/L, caused an early increase in plasma GIP and a later increase in GLP-1 concentrations (P < 0.01), suppressed appetite (P < 0.05), and reduced energy intake (P < 0.01) compared with intraduodenal infusion of saline. There was a close association between the increase in GLP-1 and decrease in appetite. Infusion of octreotide to suppress the release of gastrointestinal hormones prevented the rise in insulin, GIP, and GLP-1 induced by intraduodenal glucose infusion and reversed the suppression of appetite and reduction in energy intake. These results suggest that 1) when infused to result in plasma concentrations slightly above usual postprandial concentrations, insulin does not inhibit its own release and 2) the effects of intraduodenal glucose on appetite may be mediated through the release of GLP-1 and not insulin. PMID- 9734736 TI - Adaptive reduction in basal metabolic rate in response to food deprivation in humans: a role for feedback signals from fat stores. AB - We assessed the importance of lean and fat tissue depletion as determinants of the adaptive reduction in basal metabolic rate (BMR) in response to food deprivation by reanalyzing the data on BMR and body composition for the 32 men participating in the classic Minnesota experiment of semi-starvation and refeeding. We used individual data on BMR, body fat, and fat-free mass (FFM) assessed during the control (prestarvation) period, at weeks 12 and 24 of semistarvation (S12 and S24), and week 12 of restricted refeeding (R 12) to calculate an index of the reduction in thermogenesis at S12, S24, and R12, defined as the change in BMR adjusted for changes in FFM and fat mass, and an index of the state of depletion of the fat mass and FFM compartments at these times, defined as the deviation in fat mass or FFM relative to control values. The results indicated a positive relation between the reduction in thermogenesis and the degree of fat mass depletion (but not FFM depletion) during weight loss as well as during weight recovery (r = 0.5, P < 0.01). Furthermore, the residual variance was predicted by the initial (prestarvation) percentage fat and the cormic index (sitting height/height). Taken together, these results in normal weight men responding to severe food deprivation reveal anthropometric predictors for human interindividual variability in the capacity for energy conservation and suggest that the adaptive reduction in BMR is partly determined by an autoregulatory feedback control system linking the state of depletion of fat stores to compensatory mechanisms that suppress thermogenesis. PMID- 9734737 TI - Protein turnover and energy expenditure increase during exogenous nutrient availability in sickle cell disease. AB - In the present study, energy expenditure (EE) and rates of whole-body protein, glucose, and lipid metabolism were assessed in 8 African American sickle cell disease (SCD) patients and in 6 healthy African American control subjects during the infusion of amino acids, glucose, and lipid. Whole-body protein, glucose, and lipid kinetics were estimated by using L-[1-(13)C]leucine, D-[6,6-(2)H2]glucose, and [(2)H5]glycerol, respectively. After a 2-h tracer equilibration period and a 0.5-h basal period, nutrients were administered intravenously for 3 h with 16% of the energy as protein, 52% as carbohydrate, and 32% as fat. Breath and blood were collected during the last 30 min of nutrient infusion and EE was measured by indirect calorimetry. EE was 14% greater (P < or = 0.05) in SCD patients [145.0 +/- 3.5 kJ x kg fat-free mass (FFM)(-1) x d(-1)] than in control subjects (126.8 +/- 3.8 kJ x kg FFM(-1) x d(-1)). Whole-body protein breakdown (4.4 +/- 0.4 compared with 3.1 +/- 0.1 mg x kg FFM(-1) x min(-1), P < or = 0.05) and protein synthesis (4.6 +/- 0.4 compared with 3.2 +/- 0.1 g x kg FFM(-1) x min(-1), P < or = 0.05) were 42% and 44% greater, respectively, in the SCD patients than in control subjects, but whole-body amino acid oxidation (0.90 +/- 0.05 compared with 1.03 +/- 0.09 mg x kg FFM(-1) x min(-1)) was not significantly different between the 2 groups. Whole-body glucose and lipid kinetics did not differ significantly between the groups. EE increased in SCD patients during exogenous nutrient availability, and the additional energy required for the accelerated rates of whole-body protein breakdown and synthesis made a significant contribution to the increase in EE. These metabolic aberrations may increase the dietary energy and protein requirements of SCD patients. PMID- 9734738 TI - Increasing amounts of dietary fiber provided by foods normalizes physiologic response of the large bowel without altering calcium balance or fecal steroid excretion. AB - Nine healthy, young men consumed constant diets to determine selected large bowel, serum cholesterol and triacylglycerol, and calcium balance responses to 3 amounts of fiber provided by a mixture of fruit, vegetables, and grains. The diets, each consumed for 1 mo, contained 16, 30, and 42 g total fiber/d, of which 2.9, 4.8, and 7.7 g, respectively, was soluble. Mean daily wet and dry stool weights increased with each fiber addition. The first fiber addition increased defecation frequency and decreased fecal pH, bile acid concentration, and neutral steroid concentration; the second addition had no further effect. Mean weight of each defecation and stool moisture did not increase and serum cholesterol and triacylglycerol concentrations, calcium balance, and gastrointestinal transit time did not decrease as fiber intake increased. We conclude that 1) fiber provided by a mixed-food diet increases stool weight as effectively as does wheat or oat bran; 2) even high amounts of dietary fiber do not change transit time or defecation frequency if they are already approximately 1 and 2-3 d, respectively; 3) food patterns consistent with the food pyramid and incorporating legumes and whole grains are necessary to achieve recommended fiber intakes of 20-35 g/d, even if energy intake is > 12.55 MJ (3000 kcal); 4) soluble fiber provided by a mixture of whole foods has no effect on serum cholesterol concentrations or output of fecal bile acids; and 5) mixed-food fiber has little effect on calcium balance when calcium intakes are high (> or = 1.5 g/d). PMID- 9734739 TI - Serum retinol concentrations in children are affected by food sources of beta carotene, fat intake, and anthelmintic drug treatment. AB - The provision of vitamin A in food sources of beta-carotene is an alternative to the distribution of high-dose capsules. To examine factors that may influence the success of food-based programs, a study was carried out in Sumatra, Indonesia, of the effect of food sources of beta-carotene, extra dietary fat, and Ascaris lumbricoides infection on serum retinol concentrations in children. Meals and snacks with various amounts of beta-carotene and fat were fed at midday to children 3-6 y of age for 3 wk. Some groups of children were dewormed with the anthelmintic levamisole before the feeding period, whereas others remained infected. Results showed that the incorporation of beta-carotene sources (mainly in the form of red sweet potatoes) into the meal significantly increased serum retinol concentrations. The greatest rise in serum retinol occurred when meals contained added beta-carotene sources and added fat and the children were dewormed. Adding more fat to the meal and deworming the children caused a rise in serum retinol similar to that seen when feeding additional beta-carotene sources. Moreover, the effects of fat and deworming together were additive to the effects of additional beta-carotene sources. When the meal contained additional beta carotene sources, added fat caused a further improvement in serum retinol concentrations but only if A. lumbricoides infection was low. These studies indicated that food-based interventions in vitamin A-deficient areas might be successful and that other interventions such as increasing dietary fat concentrations and anthelmintic treatment should be considered along with increasing consumption of beta-carotene-rich food. PMID- 9734740 TI - Dietary vitamin A and prevalence of bronchial metaplasia in asbestos-exposed workers. AB - The purpose of this investigation was to examine the association between dietary intake of vitamin A in the form of retinol and provitamin A carotenoids and the prevalence of bronchial squamous metaplasia in a sample of asbestos workers from an industrial clinic. Bronchial biopsies were obtained from 49 asbestos workers. Pulmonary function testing was done and in-person interviews were conducted to estimate dietary intake of retinol and provitamin A carotenoids, tobacco exposure, and asbestos exposure. Results indicated that workers with metaplasia reported consuming a significantly lower intake of total vitamin A [2000 retinol equivalents (RE)/d] than did subjects without metaplasia (2710 RE/d, P = 0.02). Logistic regression analyses showed that higher intakes of retinol [odds ratio (OR): 0.31; 95% CI: 0.04, 2.44], provitamin A carotenoids (OR: 0.31; 95% CI: 0.03, 2.84), and total vitamin A (OR: 0.29; 95% CI: 0.03, 2.49) were associated with a nonsignificant reduction in the OR for metaplasia (highest quartile compared with lowest quartile, adjusted ORs). Current smoking (OR: 5.25; 95% CI: 0.50, 55.1) and former smoking (OR: 2.95; 95% CI: 0.31, 28.1) were associated with a nonsignificant increase in the OR for bronchial metaplasia compared with never smoking. Greater airway obstruction [decreased forced expiratory volume in 1 s (FEV1)/forced vital capacity (FVC)] was associated with an increased OR for metaplasia (OR: 2.86; 95% CI: 1.09, 7.69). These results suggest that a higher (ie, above the median) intake of vitamin A from foods decreases the risk of bronchial metaplasia in workers occupationally exposed to asbestos. PMID- 9734741 TI - Determination of discretionary salt intake in rural Guatemala and Benin to determine the iodine fortification of salt required to control iodine deficiency disorders: studies using lithium-labeled salt. AB - The use of discretionary salt, which is salt added during cooking and at the table, as a suitable vehicle for iodine intake was assessed by measuring salt consumption using the lithium-marker technique in rural areas of Guatemala and Benin. In both countries, we studied boys aged 6-12 y and their mothers. Subjects used lithium-labeled salt after all unlabeled salt was removed from their households. In Guatemala, 24-h urine samples for 9 mother-son pairs were collected at baseline and on days 7, 8, and 9 during the use of lithium-labeled salt. Total maternal salt intake averaged 5.2 +/- 1.7 g/d (mean +/- SD), of which 77 +/- 24% came from discretionary sources, whereas Guatemalan boys consumed 1.8 +/- 0.6 g salt/d, of which 72 +/- 12% came from discretionary sources. In Benin, urine collection from 13 mother-son pairs took place at baseline and on days 5 and 7. Beninese mothers had a total salt intake of 9.0 +/- 2.9 g/d and their sons had an intake of 5.7 +/- 2.8 g/d; discretionary salt contributed 52 +/- 14% and 50 +/- 13%, respectively, of total salt consumed. Therefore, fortification of household salt appears to be an appropriate method of controlling iodine deficiency in both countries, although fortification of other salt sources could be considered in Benin. PMID- 9734742 TI - Effect of supplemental beta-carotene on plasma concentrations of carotenoids, retinol, and alpha-tocopherol in humans. AB - High doses of beta-carotene, a lipid-soluble nutrient, may affect the plasma concentrations of other lipid-soluble nutrients. The purpose of this study was to assess the effects of long-term daily supplementation with beta-carotene (50 mg/d) on circulating concentrations of other carotenoids, retinol, and alpha tocopherol over time. Data were available from 259 men and women participating in the Carotene Prevention Trial, a 2-center chemoprevention trial designed to determine whether supplemental beta-carotene can prevent second malignant tumors in patients cured of an early stage cancer of the oral cavity, pharynx, or larynx. Up to 2 blood samples were obtained before the intervention (before and after a 1-mo placebo run-in), with postrandomization samples obtained at 3, 12, 24, 36, 48, and 60 mo. Supplementation with beta-carotene produced a persistent 9 to 10-fold increase in median plasma beta-carotene concentrations (225 nmol/L at baseline to 2255 nmol/L at 3 mo) and a persistent 2-fold increase in median plasma alpha-carotene concentrations (45 nmol/L at baseline to 95 nmol/L at 3 mo). Concentrations of retinol, alpha-tocopherol, lycopene, and lutein/zeaxanthin were not affected by supplemental beta-carotene. Up to 5 y of daily supplementation with beta-carotene increased circulating concentrations of alpha- and beta-carotene, but did not alter concentrations of lycopene, lutein/zeaxanthin, retinol, or alpha-tocopherol. PMID- 9734743 TI - Dietary calcium, calcium supplementation, and blood pressure in African American adolescents. AB - BACKGROUND: Intake of calcium from the diet is inversely associated with blood pressure in observational studies and animal models but randomized trials in humans have found only small effects of calcium supplementation on blood pressure. A blood pressure-lowering effect of calcium supplementation may thus be restricted to persons with a low intake of calcium from the diet and specific genetic or other characteristics. OBJECTIVE: A randomized trial was conducted to assess the effect of calcium supplementation on blood pressure in African American adolescents. Rapid growth during adolescence may increase calcium requirements, and avoidance of milk and milk products by some African Americans can result in low intake of calcium. DESIGN: One hundred sixteen adolescents (65 girls, 51 boys; mean age: 15.8 y) were given calcium (1.5 g/d) or placebo for 8 wk in a randomized, double-blind, crossover design. Blood pressure was measured after 2, 4, and 8 wk. Dietary calcium was determined with a validated food frequency questionnaire. RESULTS: The net effect (+/-SE) of calcium supplementation on diastolic blood pressure was a reduction of 1.9 +/- 1.1 mm Hg (P = 0.04, one-tailed t test). Blood pressure reduction was greater in adolescents with lower intake of calcium from the diet (P = 0.003, one-tailed t test for interaction): -4.9 +/- 1.6, -2.3 +/- 1.6, and 1.4 +/- 1.8 mm Hg for change in the lower (0.024-0.067 g Ca/MJ), middle (0.069-0.091 g Ca/MJ), and upper (0.093-0.217 g Ca/MJ) tertiles, respectively. No main effect on systolic blood pressure was detected. CONCLUSION: These findings suggest that calcium supplementation may lower diastolic blood pressure in African American adolescents with low dietary intakes of calcium. PMID- 9734744 TI - Linking maternal and infant benefits of a nutritional supplement during pregnancy and lactation. AB - To evaluate the effect of a nutritional supplement on change in women's weight during a reproductive cycle and on the difference in birth weight between one infant and the previous one, we analyzed data on 176 complete reproductive cycles from an experiment that was conducted in rural Guatemala. Women with an initial weight <50 kg were classified as marginally nourished or malnourished. Women whose intake of the supplement was in the top 2 tertiles were distinguished from those whose intake was in the lowest tertile. Linear regression modeling was used to estimate the effect of supplementation on these outcomes and to control for confounding factors. Malnourished women gained weight during the reproductive cycle, but their second (study) infant tended to weigh less at birth than their prior-born infant. Higher intakes of supplement were associated with a less negative difference in birth weight. Marginally nourished women lost weight during the reproductive cycle and their second (study) infant tended to weigh more at birth than their prior-born infant. Higher intakes of supplement were associated with a less negative weight [corrected] trend for the women themselves. Well-nourished women and their infants did not show any of these benefits from supplementation. These findings help explain past contradictory findings on maternal depletion as well as on the benefits of nutritional supplementation for mothers and their infants. PMID- 9734745 TI - Influence of smoking on vitamin E status during the third trimester of pregnancy and on breast-milk tocopherol concentrations in Spanish women. AB - Concentrations of antioxidants in breast milk probably define the degree of protection it can offer against peroxidation. The aim of the present investigation was to determine the differences in vitamin E status of Spanish women smokers and nonsmokers in their third trimester of pregnancy and the concentrations of tocopherol in their milk. Vitamin E intake was determined during the third trimester of pregnancy by using a 5-d dietary record (including a Sunday) and by recording the quantities provided by supplements. HPLC was used to determine vitamin E concentrations in subjects' serum during the third trimester, in transitional breast milk on days 13-14 of lactation, and in mature breast milk on day 40 of lactation. Subjects also answered a questionnaire about their smoking habits during pregnancy. Subjects were grouped as nonsmokers (71.9%; n = 41) or smokers (28.1%; n = 16). Although vitamin E intake was somewhat greater in nonsmokers, the difference was not significant. Ratios of vitamin E to polyunsaturated fatty acids were practically the same in both groups. The use of vitamin E supplements was limited and did not modify the results of the study. No significant differences in these serum indexes were found between smokers and nonsmokers, and no subject had deficient serum vitamin E concentrations. However, vitamin E concentrations in mature milk were significantly lower in smokers than in nonsmokers. Although it is already known that maternal smoking favors peroxidation events in newborns, if the concentration of antioxidants (vitamin E) in smokers' breast milk is also lower, it might aggravate the peroxidation problems of their newborns. PMID- 9734746 TI - Estimation of total body water in very-low-birth-weight infants by using anthropometry with and without bioelectrical impedance and H2[(18)O]. AB - The usefulness of bioelectrical impedance (BI) with anthropometry to measure total body water (TBW) was evaluated in very-low-birth-weight (VLBW) infants. A specific regression equation to measure TBW in a VLBW population was developed by simultaneously using the H2[(18)O] dilution method and BI in 12 infants with a gestational age of 24-30 wk and weighing <1200 g at birth. After an oral dose of H2[(18)O], the tracer dilution was measured in expired carbon dioxide. BI measurements were made with a model BIA-101 apparatus (RJL Systems, Detroit). Electrodes were placed in the standard position as well as proximally on the leg and the forearm. The best correlation was observed between body weight and TBW (r = 0.989). For BI, the best correlation was obtained when gestational age was used as a covariable along with body weight and crown-heel length (r = 0.985). The correlation was comparable with proximal electrode placement (r = 0.985). The new correlation was evaluated in 6 infants weighing < 1008 g. A significant correlation between BI and H2[(18)O]-measured TBW was observed (r = 0.988). Published regression equations for infants consistently gave higher estimates of TBW in another group of 14 infants weighing <1200 g than did the new correlations. TBW represented 84-95% of body weight in these VLBW infants. TBW could be computed simply from body weight alone. Use of BI and length as covariables did not add significantly to the estimate of TBW in VLBW infants. PMID- 9734747 TI - Influence of sex, seasonality, ethnicity, and geographic location on the components of total energy expenditure in young children: implications for energy requirements. AB - BACKGROUND: There are limited data on the influence of body composition, sex, seasonality, ethnicity, and geographic location on the components of energy expenditure in children. OBJECTIVE: The objective was to examine the determinants of total energy expenditure (TEE), resting energy expenditure (REE), and activity related energy expenditure (AEE) in children. DESIGN: Cross-sectional data from 232 children (4-10 y of age) from 4 ethnic groups (white American, African American, Guatemalan Mestizo, and Native American Mohawk) were examined. RESULTS: In 104 white children studied in Vermont and Alabama, TEE was significantly higher in spring than in fall, higher in boys than in girls, and higher in children in Vermont (all effects: approximately 0.42 MJ/d, P < 0.05). The significant effect of sex was explained through REE; the influences of season and location were explained through AEE. In all children, there was no effect of sex but a significant effect of ethnicity (P < 0.01) on TEE: a significant effect of sex (P < 0.01) and no effect of ethnicity (P = 0.16) on REE; and no effect of sex and a significant effect of ethnicity on AEE. The significant effects of ethnicity were due to lower values in Guatemalan children. TEE correlated most strongly with weight (r = 0.81) and fat-free mass (r = 0.79-0.81); REE with weight (r = 0.85) and fat-free mass (r = 0.80-0.87); and AEE with maximal oxygen consumption (r = 0.54), fat-free mass (r = 0.50), and fat mass (r = 0.49). CONCLUSIONS: 1) Season and location influenced TEE in children through their effects on AEE, 2) a higher REE in boys was consistent across all groups examined, 3) Guatemalan children had lower TEE due to a lower AEE, 4) body weight may be the best predictor of TEE, and 5) maximal oxygen consumption was the strongest marker of AEE. PMID- 9734748 TI - Evidence of altered central nervous system development in infants with iron deficiency anemia at 6 mo: delayed maturation of auditory brainstem responses. AB - Iron deficiency anemia has long been thought to have effects on the central nervous system (CNS). Finding direct evidence of this in human infants, however, has been challenging. Auditory brainstem responses (ABRs) provide a noninvasive means of examining an aspect of the CNS that is rapidly maturing during the age period when iron deficiency is most common. ABRs represent the progressive activation of the auditory pathway from the acoustic nerve (wave I) to the lateral lemniscus (wave V). The central conduction time (CCT, or wave I-V interpeak latency) is considered an index of CNS development because myelination of nerve fibers and maturation of synaptic relays lead to an exponential reduction in the CCT from birth to 24 mo. In 55 otherwise healthy, 6-mo-old Chilean infants (29 with iron deficiency anemia and 26 nonanemic control infants), the CCT was longer in those who had been anemic at 6 mo, with differences becoming more pronounced at 12- and 18-mo follow-ups despite effective iron therapy. The pattern of results--differences in latencies but not amplitudes, more effects on the late ABR components (waves III and V), and longer CCTs (as an overall measure of nerve conduction velocity)--suggested altered myelination as a promising explanation, consistent with recent laboratory work documenting iron's essential role in myelin formation and maintenance. This study shows that iron deficiency anemia in 6-mo-old infants is associated with adverse effects on at least one aspect of CNS development and suggests the fruitfulness of studying other processes that are rapidly myelinating during the first 2 y of life. PMID- 9734749 TI - Stress response in school-age children who have been growth retarded since early childhood. AB - BACKGROUND: Approximately 39% of children aged <5 y in developing countries are growth retarded (stunted) and many have poor mental development and behavioral abnormalities. Animal research suggests that an altered stress response may contribute to the negative outcomes following undernutrition. OBJECTIVE: We tested the hypothesis that stunted children have higher salivary cortisol concentrations and heart rates and altered behavior when compared with nonstunted children when social background was controlled for. DESIGN: We compared 30 stunted with 24 nonstunted children, all of whom were 8-10 y old and lived in the same poor areas of Kingston, Jamaica. All subjects were participants in a prospective, longitudinal, case-control study of children who were stunted in early childhood. We administered a test session (including psychologic and physical stressors), measured baseline and response levels of salivary cortisol and heart rate, and observed behavior. RESULTS: Compared with nonstunted children, stunted children had higher salivary cortisol concentrations (P = 0.007), had higher heart rates during the psychologic test session (P = 0.03), exhibited enhanced cardiovascular responsivity to a physical stressor (P = 0.04), vocalized less, were more inhibited, and were less attentive. After birth weight or social background and maternal and child intelligence quotients were controlled for, the differences in cortisol concentration and cardiovascular reactivity remained significant. CONCLUSIONS: Our findings suggest that consistent growth retardation since early childhood affects physiologic arousal, which, we speculate, may contribute to the poor cognitive functioning and immune responses of stunted children and the relation between adult short stature and increased cardiovascular risk. PMID- 9734750 TI - Interaction between vitamin K nutriture and bacterial overgrowth in hypochlorhydria induced by omeprazole. AB - Subjects taking a hydrogen pump blocking agent (omeprazole) develop bacterial overgrowth of the small intestine. We tested the hypothesis that this bacterial overgrowth produces menaquinones, which would meet the vitamin requirement in situations of vitamin K deficiency. In a crossover-type design, 13 healthy volunteers eating a phylloquinone-restricted diet for 35 d were randomly assigned to take omeprazole during the first period of study or starting on day 15 until the end of the study. Coagulation times, serum osteocalcin [total osteocalcin and undercarboxylated osteocalcin (ucOC)], plasma phylloquinone, urinary gamma carboxyglutamic acid, and plasma undercarboxylated prothrombin (PIVKA-II) were measured. Plasma phylloquinone concentrations declined 82% with dietary phylloquinone restriction (P < 0.05) and were not significantly different in the period when the diet was combined with omeprazole treatment (P > 0.05). The mean value for PIVKA-II during the phylloquinone-restricted diet significantly increased 5.7-fold from baseline (P < 0.05); however, the combination of omeprazole treatment and the phylloquinone-restricted diet significantly reduced PIVKA-II values by 21% (P < 0.05) compared with the diet period alone. There were no alterations in total or percentage ucOC concentrations during the phylloquinone-restricted diet or during the period of diet plus omeprazole treatment. Our data support the hypothesis that bacterial overgrowth results in the synthesis and absorption of menaquinones. These menaquinones contribute to vitamin K nutriture during dietary phylloquinone restriction, but not enough to restore normal vitamin K status. PMID- 9734751 TI - Small-intestinal digestion of partially resistant cornstarch in healthy subjects. AB - The aims of this study were to measure the amount of starch from partially resistant starches (retrograded and complexed high-amylose cornstarches) escaping small-intestinal digestion in healthy humans by use of an intubation method and to compare these data with data obtained by indirect in vitro methods. Experiments were carried out in vivo in 6 healthy humans by using ileal intubation and stool analysis and in vitro by using 3 different methods for analyzing resistant starch. In intubated subjects, 51 +/- 2% of the retrograded and 21 +/- 2% of the complexed starch was delivered to the ileum and was fermented almost completely in the colon. In vitro estimates of the absorption of resistant starch were much lower. We conclude that technologically modified starches may substantially increase the amount of carbohydrate available for colonic fermentation in humans, but that in vitro measurements of resistant starch are inaccurate for predicting malabsorption in healthy humans. PMID- 9734752 TI - Mechanisms by which wheat bran and oat bran increase stool weight in humans. AB - Generally, stool weight is significantly increased by adding sources of insoluble fiber to the diet. Comparable amounts of fiber provided by wheat and oat brans have the same effect on daily stool output, even though > 90% of wheat bran fiber but only 50-60% of oat bran fiber is insoluble. To determine the bases for these increases in stool weight, stool samples collected from 5 men in 2 constant diet studies that determined the effects of wheat and oat brans on large-bowel physiology were fractionated by using a physicochemical procedure into plant, bacterial, and soluble fractions, which were weighed and analyzed for sugar content and composition. Nitrogen, crude fat, and ash outputs were also determined. Wheat bran increased the fecal concentration of sugars and mass of plant material more than did oat bran, whereas oat bran increased fecal bacterial mass more. Each fiber source increased nitrogen, ash, and fat excretion, but excretion of fat was greater with oat bran. The apparent digestibility of plant derived neutral sugars decreased significantly when wheat but not oat bran was consumed. The apparent digestibility of neutral sugars provided by wheat bran was 56%; the apparent digestibility of those provided by oat bran was 96%. We conclude that bacteria and lipids are major contributors to the increase in stool weight with oat bran consumption, whereas undigested plant fiber is responsible for much of the increase in stool weight with wheat bran consumption. Results are consistent with the hypothesis that oat bran increases stool weight by providing rapidly fermented soluble fiber in the proximal colon for bacterial growth, which is sustained until excretion by fermentation of the insoluble fiber. PMID- 9734753 TI - Determinants of increased energy expenditure in HIV-infected women. AB - BACKGROUND: Little is known about sex-specific effects of HIV infection on energy expenditure. OBJECTIVE: We investigated the determinants of energy expenditure in HIV-infected women. DESIGN: Resting energy expenditure (REE), body composition, and hormonal and nutritional indexes were compared in 33 ambulatory, premenopausal HIV-infected women and 26 weight-matched, healthy premenopausal control subjects. REE was determined by indirect calorimetry and body composition by dual-energy X-ray absorptiometry (DXA), bioelectrical impedance analysis, and skinfold-thickness analysis. Hormonal indexes included leptin, testosterone, estradiol, and insulin-like growth factor I. RESULTS: HIV-infected subjects had a higher REE than control subjects [6794 +/- 1374 compared with 6011 +/- 607 kJ/d (1624 +/- 329 compared with 1437 +/- 145 kcal/d), P = 0.0096]. On average, REE was 119 +/- 23% of Harris-Benedict predictions in HIV-infected subjects compared with 102 +/- 9% for control subjects (P = 0.0007). In HIV-infected subjects, REE was highly correlated with fat-free mass (FFM) by DXA (R = 0.641, P < 0.001), but not with weight or disease status. The slope of the regression equation for REE and FFM was significantly greater (P = 0.027, analysis of covariance) for HIV infected subjects [REE (kJ/d) = 203.5 (kg FFM) - 1237] than for control subjects [REE (kJ/d) = 77.4 (kg FFM) + 2923]. In a stepwise regression analysis, FFM was the most significant variable (P = 0.005), followed by free testosterone (P = 0.029), which together explained 49% of the variation in REE. The final equation was REE (kJ/d) = 230.8 (kg FFM) + 395.9 (free testosterone, pmol/L) - 3304. CONCLUSIONS: Energy expenditure was higher in HIV-infected women than in control women. FFM is the primary determinant of REE in HIV-infected women, but energy expenditure is greater per kg FFM in HIV-infected subjects than in control subjects, which may contribute to the wasting syndrome. PMID- 9734754 TI - Provitamin A carotenoid intake and carotid artery plaques: the Atherosclerosis Risk in Communities Study. AB - We examined the cross-sectional association between intake of carotenoids with provitamin A activity and carotid artery plaques in 12,773 participants of the Atherosclerosis Risk in Communities Study aged 45-64 y. Usual diet was assessed with a 66-item food-frequency questionnaire. Plaques were examined by B-mode ultrasound of multiple carotid artery segments. In both women and men, those in the highest quintile of carotenoid consumption had a lower prevalence of plaques (women, 25.4%; men, 36.0%) than those in the lowest quintile of carotenoid consumption (women, 29.3%; men, 39.8%). The prevalence odds ratios contrasting extreme intake quintiles were 0.82 (95% CI: 0.70, 0.97) in women and 0.85 (95% CI: 0.72, 1.01) in men. The associations diminished slightly after potential confounders were adjusted for. In women, the inverse association was particularly strong for current smokers (adjusted odds ratio contrasting extreme quintiles: 0.67; 95% CI: 0.45, 0.98). In men, no such effect modification by smoking was seen. The inverse association was somewhat stronger in men aged 55-64 y than in those aged 45-54 y, whereas age made little difference in women. These findings, together with previous findings that carotenoid intake was unrelated to average carotid artery wall thickness, suggest that carotenoids may exert their influence later rather than earlier in the atherosclerotic process, and support the hypothesis that carotenoids or other plant-derived compounds may play a role in preventing arterial plaque formation. PMID- 9734755 TI - Relation of anthropometry to malaria morbidity and immunity in Papua New Guinean children. AB - The interaction between malnutrition and malaria is complex and there is evidence that malnutrition decreases the susceptibility to malaria. To investigate the relation between anthropometric measurements and subsequent malaria morbidity and to examine whether the effect observed was due to interaction with host immunity, we followed for 1 y a cohort of 136 children aged 10 to < 120 mo in Wosera, East Sepik Province, Papua New Guinea. At baseline, 21% were stunted, 10% were wasted, and 5% were both stunted and wasted. After adjustment for age and use of bed nets, height-for-age z score (HAZ) at baseline predicted the number of clinical episodes of falciparum malaria during the following year: incidence rate increased with increasing HAZ. Humoral responses to specific malarial antigens were lowest in the wasted children. The prevalence of lymphoproliferative responders was not significantly different between well-nourished and undernourished children. In contrast, the prevalence of cytokine producers was higher in the undernourished than in the well-nourished children. Our findings support the view that stunting but not wasting protects against falciparum malaria. The mechanism may be related to an improved ability of malnourished children to produce certain cytokines in response to stimulation by specific malarial antigens. PMID- 9734756 TI - Zinc and rehabilitation from severe protein-energy malnutrition: higher-dose regimens are associated with increased mortality. AB - A randomized, double-blind trial was undertaken to measure the effects of zinc supplementation on catch-up growth in severe protein-energy malnutrition, with particular reference to linear growth. One hundred forty-one children between the ages of 6 mo and 3 y were enrolled after admission to a nutritional rehabilitation unit in Dhaka, Bangladesh, and randomly assigned to receive elemental zinc by mouth, 1.5 mg/kg for 15 d, 6.0 mg/kg for 15 d, or 6.0 mg/kg for 30 d, and thereafter they were followed for a total of 90 d. Anthropometric outcome measures included change in knee-heel length, midupper arm circumference, subscapular and triceps skinfold thicknesses, and change in height-for-age, weight-for-age, and weight-for-height z scores. Higher zinc doses were not associated with significant change in any anthropometric measurement, but mortality was significantly greater in children who received high-dose zinc (6.0 mg/kg) initially as opposed to those who received low-dose zinc supplementation (1.5 mg/kg) (Yates-corrected chi-square P value of 0.033 and a risk ratio of 4.53; 95% CI: 1.09 < risk ratio < 18.8). We conclude that there is no benefit to using high-dose zinc supplementation regimens and that they could contribute to increased mortality in severely malnourished children. PMID- 9734757 TI - Dietary calcium, protein, and phosphorus are related to bone mineral density and content in young women. AB - BACKGROUND: Dietary factors have been implicated in modifying bone health, although the results remain controversial, particularly in young women. OBJECTIVE: The objective of the study was to determine relations of selected dietary factors and anthropometric measurements to bone mineral density (BMD) of the spine, femoral neck, trochanter, Ward's triangle, radius, and total body and the bone mineral content (BMC) of the spine, radius, and total body. DESIGN: The study was a cross-sectional analysis of 215 women aged 18-31 y. RESULTS: Weight, height, and lean mass were correlated with bone mineral measures at every site (r = 0.17-0.78). Postmenarcheal age (years since onset of menses) was positively correlated with total-body BMD and BMC, radius BMD and BMC, and spine BMC, and negatively correlated with Ward's triangle BMD. Radius BMD was correlated with protein, calcium, and phosphorus intakes, and spine BMD and BMC were correlated with energy, protein, calcium, and phosphorus intakes. These correlations remained significant when postmenarcheal age, lean mass, and fat mass were controlled. A pattern emerged in multiple regression analyses that showed a complex relation among calcium, protein or phosphorus, and the calcium-protein or calcium-phosphorus ratio and spine or total-body BMC and BMD. All 3 variables (calcium, protein or phosphorus, and calcium-protein or calcium-phosphorus ratio) were required in the model for significance. CONCLUSIONS: Anthropometric measures were predictors of bone mass. A single ratio of calcium to phosphorus or protein did not optimize bone mass across the range of calcium intakes. PMID- 9734758 TI - Are serum leptin correlations influenced by fat placement? PMID- 9734759 TI - Biomarkers of vegetable and fruit intakes. PMID- 9734760 TI - Anorexia of aging. PMID- 9734761 TI - Do you get the picture? PMID- 9734762 TI - Osteoarthritis and synovitis as major pathoses of the temporomandibular joint: comparison of clinical diagnosis with arthroscopic morphology. AB - PURPOSE: The purposes of this investigation were to determine how common osteoarthritis and synovitis are in patients with severe, recalcitrant temporomandibular joint (TMJ) symptoms using clinical diagnostic criteria as well as arthroscopic examination, and to compare the accuracy of the clinical and arthroscopic diagnoses with respect to specificity and sensitivity. PATIENTS AND METHODS: Clinical and arthroscopic diagnoses were established in 126 joints of 84 patients with severe TMJ symptoms recalcitrant to conservative therapy. All joints were classified as having osteoarthritis (OA) or no osteoarthritis (non OA) and synovitis (syn) or no synovitis (non-syn) using clinical and arthroscopic criteria. Chi-squared analysis was used to determine whether there was a relationship between the clinical and arthroscopic diagnoses. Preoperative clinical diagnoses were compared with arthroscopic morphologic diagnoses to determine the specificity and sensitivity of the clinical diagnostic criteria for synovitis and osteoarthritis. RESULTS: A preoperative clinical diagnosis of OA was established in 59 of 126 joints (47%) compared with an arthroscopic diagnoses of OA in 82 of 126 joints (65%). Chi-squared analysis showed a significant relationship between the clinical and arthroscopic diagnosis of OA. A clinical diagnosis of OA was associated with a high specificity (.977); however, there were 23 of 82 (.293) false-negative findings and a sensitivity of only .707. A preoperative clinical diagnosis of synovitis was established in 114 of 126 joints (90%), compared with an arthroscopic diagnosis of synovitis in 112 of 126 (89%). Chi-squared analysis did not show a significant relationship between the clinical and arthroscopic diagnosis of synovitis. A clinical diagnosis of synovitis was associated with a high sensitivity (.920); however, there were 11 of 14 false positive findings (.786) associated with a low specificity (.214). CONCLUSIONS: Although there was high specificity for the clinical diagnosis of OA, the sensitivity was very low. (Comparison of clinical and arthroscopic diagnoses showed that osteoarthritis frequently escapes clinical detection. The clinical diagnosis of synovitis showed that low specificity and symptoms may be caused by other pathoses. PMID- 9734763 TI - Stability of Le Fort I osteotomy with advancement: a comparison of single maxillary surgery and a two-jaw procedure. AB - PURPOSE: This study compared single maxillary surgery and a two-jaw procedure in patients who underwent one-piece Le Fort I advancement without bone grafting. PATIENTS AND METHODS: Fifty-three patients had Le Fort I osteotomy performed using a standard technique. Twenty-two patients had maxillary surgery alone, and 31 patients additionally had a bilateral sagittal split ramus osteotomy performed. Both rigid and nonrigid fixation were used. The postoperative movement of the maxilla was investigated, comparing cephalograms taken preoperatively, 2 to 3 days postoperatively, and at least 6 months postoperatively. A computer program was used to superimpose the three radiographs. RESULTS: No difference in postoperative stability was found when the two surgical procedures were compared, and no correlation between magnitude of advancement and degree of relapse could be identified (P > .05). Nonrigid fixation in patients receiving only maxillary surgery resulted in greater postoperative forward movement of the maxilla (P = .022). CONCLUSION: This study indicates that postoperative stability of the maxilla in a two-jaw procedure is equivalent to that of single maxillary surgery. Nonrigid fixation in single maxillary surgery reduces the need for postoperative orthodontics. PMID- 9734764 TI - Comparison of morbidity of outpatient general anesthesia administered by the intravenous or inhalation route. AB - PURPOSE: This study compares the morbidity between subjects receiving general anesthesia either by an intravenous or inhalation route for the extraction of impacted third molars in an outpatient setting. PATIENTS AND MATERIALS: Forty ASA Class I subjects, 21 females and 19 males (age range, 17 to 43 years), who presented for the extraction of four impacted third molars, were studied. Subjects were alternately assigned to receive general anesthesia either by the intravenous route (group I) or the inhalation route via an endotracheal tube (group II). The parameters for comparison included psychomotor recovery, cardiovascular changes 20% above or below baseline, the frequency of nausea and vomiting perioperatively and at 48 hours, occurrence of laryngospasm and bronchospasm, the frequency of sore throat both perioperatively and at 48 hours, procedure time, and recovery time. The Trieger dot test was administered to patients at three different intervals to evaluate psychomotor recovery. All parameters were recorded for each subject and compared both within and between groups. RESULTS: There was no statistical difference found between groups I and II with regard to psychomotor recovery, the frequency of nausea and vomiting, bronchospasm, laryngospasm, or median recovery time (P < .05). However, there was greater variability in both elevation and depression of blood pressure from baseline in the intubated subjects (P < .05). These deviations were both expected and easily managed. The probability of sore throat was greater in the intubated subjects (P < .05) than the nonintubated subjects. Procedure time, although a weak association, was nonetheless found to be significantly greater for the intubated group than for the intravenous group (P < .05). SUMMARY: The results show greater cardiovascular variability, increased probability of sore throat, and slightly lengthened procedure time with the administration of general anesthesia by an inhalational route via an endotracheal intubation. However, there was no difference with regard to psychomotor recovery, recovery time, the probability of nausea and vomiting, or incidence of laryngospasm or bronchospasm. PMID- 9734765 TI - Laser-assisted endoscopic forehead lift. AB - The periorbital area is one of the most expressive areas of the face, and there are many techniques available that can be used to alter the position of the eyebrows. Traditional surgical browlift techniques use coronal, midforehead, and direct approaches. This article discusses one of the most recent innovations in forehead lifting, the laser-assisted endoscopic forehead lift. A review of the literature describes the numerous available surgical techniques used to change the position of the eyebrow. The surgical technique for the laser-assisted endoscopic forehead lift is then presented in detail and illustrated with the results of two cases. PMID- 9734766 TI - Incremental bolus versus a continuous infusion of propofol for deep sedation/general anesthesia during dentoalveolar surgery. AB - PURPOSE: This article compared the use of the traditional incremental bolus technique with the continuous infusion technique for the administration of propofol for deep sedation/general anesthesia. PATIENTS AND METHODS: Patients were sedated with midazolam and fentanyl and then had maintenance of an anesthetic state achieved with propofol administered by either of the two techniques. Data were collected to evaluate the overall surgical/anesthetic procedure, movement of the patient, and his or her hemodynamic status. RESULTS: Both groups received a mean maintenance dose of propofol exceeding 6 mg/kg/hr. However, the patients in the continuous infusion group received a statistically greater maintenance dose (continuous infusion + supplemental vs incremental bolus). All patients were maintained in a deep sedation/general anesthetic state. Respiratory and blood pressure values were comparable in both groups. However, the continuous infusion group showed improved hemodynamic stability manifested as fewer fluctuations in heart rate. Visual analog scale (VAS) questionnaires completed by the surgeon and surgical assistant reported less patient movement and improved surgical/anesthetic conditions with the continuous infusion technique. Recovery of the two groups was comparable. CONCLUSION: This study, although finding advantages in the continuous infusion technique, showed satisfactory conditions associated with both techniques. PMID- 9734767 TI - Comparison between the rigidity of bicortical screws and a miniplate for fixation of a mandibular setback after a simulated bilateral sagittal split osteotomy. AB - PURPOSE: This investigation compared the biomechanical stability of three bicortical screws with that of a single four-hole miniplate after 5-mm mandibular setback after a bilateral sagittal split osteotomy (BSSO) in cadaver mandibles. MATERIALS AND METHODS: Thirty human cadaver hemimandibles underwent BSSO followed by two different rigid fixation techniques. All specimens had no third molar, bony pathology, or evidence of mandibular fracture, and there was no history of renal disease or hyperparathyroidism. The specimens were randomly divided into two groups. In group I, three bicortical screws were placed at the superior border, and in group II, one four-hole miniplate was secured on the external oblique ridge with four monocortical screws. The bony height of the mandible was recorded. Maximum resistance load (MRL), the greatest load recorded just before a sudden decrease in load level (bone or fixation failure), was recorded when the mandibles were tested in a compression machine. Multiple regression analysis was used to evaluate the differences in bone height and the MRL between groups I and II. RESULTS: The mean bone height in groups I and II were 28.64 +/- 2.50 mm and 28.72 +/- 4.08 mm, respectively. The mean MRL in group I (20.49 +/- 7.22 kg) was greater than in group II (17.41 +/- 7.81 kg). The multiple regression analysis showed no significant difference in the bone height and the MRL between group I and group II (beta = 2.3492, P = .4114). CONCLUSION: There was no statistically significant difference in stability provided the two techniques. PMID- 9734768 TI - Interleukin-1beta in synovial fluid from the arthritic temporomandibular joint and its relation to pain, mobility, and anterior open bite. AB - PURPOSE: The purpose of this study was to investigate whether interleukin-1beta in synovial fluid or blood plasma is involved in the development of pain or hyperalgesia of the temporomandibular joint (TMJ), as well as reduced mandibular mobility and anterior open bite. PATIENTS AND METHODS: Twenty-nine patients with TMJ arthritis and seven healthy subjects were studied. VAS measurement of TMJ tenderness on palpation of the TMJ (TDP), TMJ pressure pain threshold and tolerance level (PPTL), mandibular mobility, pain during joint movements, and degree of anterior open bite (AOB) were assessed. IL-1beta levels were analyzed in TMJ synovial fluid (SF-IL-1beta) and blood samples and correlated with the preceding factors. RESULTS: SF-IL-1beta showed significant positive correlations with VAS measurement of pain, TDP, and AOB and a negative correlation with PPTL. CONCLUSIONS: This study indicates that IL-1beta in the synovial fluid is associated with pain and hyperalgesia in the TMJ region as well as an anterior open bite. Concerning the latter condition, IL-1beta seems to be a warning signal of tissue destruction. PMID- 9734770 TI - The use of nonopioid drugs in management of chronic orofacial pain. AB - Although controversial, opioid analgesics have been prescribed for patients with chronic facial pain. Based primarily on survey data and a few well-controlled clinical trials, long-term opioid treatment provides adequate pain reduction in 41% to 100% of patients with chronic nonmalignant pain. However, only 25% of chronic facial pain patients reported adequate pain relief with chronic opioid treatment. Work, home, and school function are generally reestablished or maintained during chronic opioid treatment, but 25% to 38% of patients remain dysfunctional, and one study indicated that 20% of patients became dysfunctional during treatment. Chronic opioid treatment is associated with many transient side effects; constipation, dizziness, nausea, vomiting, itching, and fatigue have been reported in 5% to 42% of patients taking opioids over 1 year. Although survey studies suggest that the risks of addiction are low in typical patients, drug abuse rates up to 17.3% and prescription abuse rates up to 27.6% were reported within groups of chronic opioid users. Chronic opioid use induces analgesic tolerance and physical dependence, which may result in a serious abstinence syndrome in users and children born to users. Chronic opioid use also may induce harmful immune system changes, diminish cognitive and motor function, and produce nociceptive hyperexcitability. This article shows that the use of long-term opioids for chronic facial pain is not justified based on the available data. Despite these perceived problems, there is anecdotal evidence that chronic facial pain patients will respond positively to opioid analgesics. In our experience, the pain assessment scale and a modification of the World Health Organization's three-step analgesic ladder, which prescribes nonopioid analgesics, can be the starting point for the successful management of chronic facial pain. PMID- 9734769 TI - A comparative study of osseointegration of titanium implants in autogenous and freeze-dried bone grafts. AB - PURPOSE: This study was undertaken to compare the rate and degree of osseointegration of dental implants when placed into either autogenous corticocancellous chip or freeze-dried corticocancellous chip bone grafts. MATERIALS AND METHODS: The canine ilium was used as the model site. Thirty experimental and 15 control implants were placed in 15 dogs: autogenous versus freeze-dried corticocancellous chip bone grafts around the exposed implant surfaces. In addition to the placement of control implants, the apical portion of the grafted implants acted as their own control. The implants were harvested at 1, 2, and 3 months. The evaluation of the integration process was performed by means of light microscopy, microradiography, and histomorphometry. RESULTS: Using this model, the results indicate that at 1 month there was no statistical difference in the degree of osseointegration in the two bone grafts. At 2 months, there was a statistically greater degree of osseointegration noted in the autogenous corticocancellous chip sites than in the freeze-dried bone grafts. At 3 months, the degree of osseointegration in the two groups was 70% and 33%, respectively. At 3 months, there was virtually 100% integration with trabecular bone at the control implant sites. CONCLUSION: The results indicate that at 2 months postoperatively implants placed in an autogenous bone chip graft osseointegrate to a significantly greater degree than implants placed in a freeze dried bone chip graft, and this difference remains at 3 months. PMID- 9734771 TI - The use of opioid drugs in management of chronic orofacial pain. AB - The use of opioid analgesics for the management of patients with chronic pain is controversial. However, randomized and double-blind clinical trials have shown that in select groups of patients with chronic pain, the daily administration of oral opioids decreases pain levels and improves quality of life. This article provides a review of the most recent basic and clinical research supporting the rationale for the use of opioids in a select group of patients with chronic orofacial pain. Critical to the employment of this technique are proper patient evaluation and use of comprehensive management strategies. This management scheme should be reserved for patients with chronic pain that is refractory to most nonopioid therapy. The primary reason for the clinician's reluctance to initiate long-term opioid therapy for their patients with chronic pain is the potential risk of developing opioid tolerance, dependence, or addiction. In contrast to these beliefs, studies have shown a nonexistent to low risk of opioid dependence or addiction behavior with administration of scheduled oral opioids in chronic pain patients. It is essential that potential patients for this type of therapy have been carefully screened and have not had a history of drug addiction. The criteria to be evaluated when considering opioid therapy for chronic orofacial pain control include 1) inadequate pain diminution from prior nonopioid therapy, 2) negative history of substance abuse, 3) definitive determination that the pain being treated is of physiologic rather than psychologic origin, 4) a willingness to adhere to an "opioid contract" between the doctor and patient, 5) compliance with a scheduled, rather than "as needed" or "breakthrough," administration of an oral opioid, and 6) close clinical follow-up to evaluate pain relief, return to daily activities, and titration of drug levels. If these criteria are followed, administration of oral opioids may be a successful means of decreasing the patient's debilitating chronic pain to tolerable levels, enabling an improvement in the quality of life and return to function. PMID- 9734772 TI - Radiolucent lesion of the mandibular angle and ramus. PMID- 9734774 TI - Acute actinomycosis presenting as an ulcerated palatal mass. PMID- 9734773 TI - Aspiration and ingestion of foreign bodies in oral and maxillofacial surgery: a review of the literature and report of five cases. PMID- 9734775 TI - Leiomyosarcoma of the maxilla: a case report. PMID- 9734776 TI - Arthroscopic laser debridement of temporomandibular joint fibrous and bony ankylosis: case report. PMID- 9734777 TI - Li-Fraumeni syndrome and osteosarcoma of the maxilla. PMID- 9734778 TI - Tonsillar hemangiopericytoma versus lymphoma in AIDS patients. PMID- 9734779 TI - Risks of using platelet rich plasma gel. PMID- 9734780 TI - Regulatory mechanisms in expression of the traY-I operon of sex factor plasmid R100: involvement of traJ and traY gene products. AB - BACKGROUND: The plasmid R100 encodes tra genes essential for conjugal DNA transfer in Escherichia coli. Genetic evidence suggests that the traJ gene encodes a positive regulator for the traY-I operon, which includes almost all the tra genes located downstream of traJ. The molecular mechanism of regulation by TraJ, however, is not yet understood. traY is the most proximal gene in the traY I operon. TraY promotes DNA transfer by binding to a site, sbyA, near the origin of transfer. TraY is suggested to have another role in regulation of the traY-I operon, since it binds to two other sites, named sbyB and sbyC, located in the region preceding traY-I. RESULTS: Using a traY-lacZ fusion gene, we showed that the traY-I operon was expressed only in the presence of traJ. The TraJ-dependent expression of traY-I required the E. coli arcA gene, which encodes a host factor required for conjugation. TraJ-dependent transcription occurred from a promoter (named pY) located upstream of traY-I. The isolated TraJ protein was found to bind to a dyad symmetry sequence, named sbj (specific binding site of TraJ), which existed in the intergenic region between traJ and traY-I. We also demonstrated that TraY repressed the TraJ-dependent expression of traY-I at the TraY binding sites, sbyB and sbyC, which overlapped with pY. CONCLUSIONS: TraJ is a protein which binds to the sbj site in the region upstream of the promoter pY and positively regulates expression of the traY-I operon in the presence of the E. coli arcA gene. Since sbj is located 93bp upstream of pY in the intergenic region between traJ and traY-I, TraJ presumably contacts with a transcription apparatus to promote transcription from pY. TraY, which is known to activate the initiation of conjugal DNA transfer, has a new role in the transcriptional autoregulation of traY-I expression. At levels which are sufficient to initiate conjugal DNA transfer, TraY represses traY-I transcription in the presence of TraJ. PMID- 9734781 TI - Defect in cytokinesis of fission yeast induced by mutation in the WD40 repeat motif of a TFIID subunit. AB - BACKGROUND: TBP-associated factors contain a variety of structural motifs and their related in vivo significance has remained unclear. We have attempted to identify specific biological phenomena linked to a particular domain of a TAF by analysing domain-exchanged chimeric mutants between Schizosaccharomyces pombe (Sp) and Saccharomyces cerevisiae (Sc) counterparts. RESULTS: Contrary to the case of TBP, Sp TAF containing the WD40 repeat cannot be exchanged for its Sc counterpart, despite their highly conserved primary structures. This 'species specific' function locates in the N-terminal region. The C-terminal region, largely consisting of the WD40 repeat, is exchangeable for the corresponding region of its Sc counterpart. Growth of the strain harbouring this C-terminal chimeric mutant is temperature-sensitive. The chimeric gene product did not disappear at a restrictive temperature, a finding which strongly suggests that the growth defect is caused by an aberration in the interactions through the WD40 repeat structural motif. With temperature elevation, the chimeric mutants underwent drastic morphological changes due to a defect in cytokinesis. CONCLUSIONS: The WD40 repeat of TAF is primarily involved in reactions which might regulate cytokinesis in Sp. PMID- 9734782 TI - Functional sites of human PCNA which interact with p21 (Cip1/Waf1), DNA polymerase delta and replication factor C. AB - BACKGROUND: PCNA, an eukaryotic DNA sliding clamp interacts with replication factors and the cell cycle protein, p21(Cip1/Waf1) and functions as a molecular switch for DNA elongation. To understand how DNA replication is regulated through PCNA, elucidation of the precise mechanisms of these protein interactions is necessary. RESULTS: Loop-region mutants in which human PCNA sequences were substituted with the corresponding Saccharomyces cerevisiae PCNA regions were prepared. Analysis of their functions, along with previously prepared alanine scanning mutants, demonstrated that some loops interact with DNA polymerase delta (pol delta) and replication factor C (RFC). The p21 binding sites of PCNA, mapped by affinity measurement of the mutant forms, found to be located within a distinct structure of the PCNA monomer, overlap with RFC- and pol delta interaction sites. Competition between p21 and pol delta or RFC for binding to PCNA results in efficient inhibition of its stimulation of pol delta DNA synthesis and RFC ATPase but not of PCNA loading on DNA by RFC. CONCLUSIONS: Semi saturated amounts of p21 selectively block formation of the active pol delta complex but not the RFC-PCNA complex at 3'-ends of DNA primers. This differential effect may explain the specific inhibition of DNA replication by p21. PMID- 9734783 TI - A new sigma factor, SigD, essential for stationary phase is also required for multicellular differentiation in Myxococcus xanthus. AB - BACKGROUND: Myxococcus xanthus is a gram-negative bacterium that undergoes spectacular development to form multicellular fruiting bodies under nutrient deprivation. Inside a fruiting body, vegetative cells differentiate into spores. A number of sigma factors have been shown to play roles in the regulation of gene expression in the M. xanthus life cycle. Additional sigma factors were searched to further explore the M. xanthus life cycle. RESULTS: A new sigma factor was identified, SigD, which consists of 297 amino acid residues. Two transcription initiation sites for the sigD gene were detected by primer extension analysis using total RNA from the vegetative and developmental cells, one of which was specific for development. The characterization of sigD-lacZ fusion strains demonstrated that sigD expression increased during entry into stationary phase of vegetative growth and during early development. A deletion mutant of sigD exhibited growth defects during the late-log phase and stationary phase, with dramatically reduced cell viability. The patterns of protein synthesis at late log phase of vegetative growth and at early development on CF agar plates were significantly different between the deletion mutant and the wild-type strain. The deletion mutant was more sensitive to various stresses when compared with the wild-type strain and did not accumulate trehalose in response to osmotic stress. The deletion mutant also showed a significant delay in fruiting body formation and sporulation and yielded fewer spores than the wild-type strain. CONCLUSIONS: SigD shows characteristic features of the stationary phase sigma factors and also plays important roles in multicellular differentiation of M. xanthus. PMID- 9734784 TI - Smad6 functions as an intracellular antagonist of some TGF-beta family members during Xenopus embryogenesis. AB - BACKGROUND: Bone morphogenetic proteins (BMPs) transmit signals via the intracellular protein Smad1, which is phosphorylated by ligand bound receptors, translocates to the nucleus, and functions to activate BMP target genes. Recently, a subclass of Smad proteins has been shown to inhibit, rather than transduce, BMP signalling, either by binding to the intracellular domain of BMP receptors, thereby preventing phosphorylation-mediated activation of Smad1, or by binding directly to Smad1, thereby inhibiting its ability to activate gene transcription. RESULTS: We have identified a Xenopus Smad (Smad6) that is 52% identical to mammalian Smad6, an inhibitory Smad. The spatial pattern of expression of Smad6 changes dynamically during embryogenesis and is similar to that of BMP-4 at the tailbud stage. Overexpression of Smad6 in Xenopus embryos phenocopies the effect of blocking BMP-4 signalling, leading to dorsalization of mesoderm and neuralization of ectoderm. Xenopus Smad6 completely blocks the activity of exogenous BMP-4, and, unlike human Smad6, partially blocks the activity of activin, in a mesoderm induction assay. We also find that Smad6 protein accumulates at the membrane in some cells but is partially or completely restricted to nuclei of most overexpressing cells. CONCLUSIONS: We have identified an inhibitory Xenopus Smad, Smad6, that functions as an intracellular antagonist of activin and BMP-4 signalling. Our finding that Smad6 protein is partially or completely restricted to nuclei of most overexpressing cells suggests that it may employ a novel or additional mechanism of action to antagonize TGF-beta family signalling other than that reported for other inhibitory Smads. PMID- 9734785 TI - Axin, an inhibitor of the Wnt signalling pathway, interacts with beta-catenin, GSK-3beta and APC and reduces the beta-catenin level. AB - BACKGROUND: The Wnt/Wingless signalling pathway plays an important role in both embryonic development and tumorigenesis. Beta-catenin and Axin are positive and negative effectors of the Wnt signalling pathway, respectively. RESULTS: We found that Axin interacts with beta-catenin and glycogen synthase kinase-3beta (GSK 3beta). Furthermore, the regulation of the G-protein signalling (RGS) domain of Axin is associated with the colorectal tumour suppressor adenomatous polyposis coli (APC). Overexpression of Axin in the human colorectal cancer cell line SW480 induced a drastic reduction in the level of -catenin. Interaction with beta catenin and GSK-3beta was required for the Axin-mediated beta-catenin reduction. CONCLUSION: Axin interacts with beta-catenin, GSK-3beta and APC, and negatively regulates the Wnt signalling pathway, presumably by regulating the level of beta catenin. PMID- 9734786 TI - Using an office system intervention to increase breast cancer screening. AB - OBJECTIVE: To evaluate an innovative approach to continuing medical education, an outreach intervention designed to improve performance rates of breast cancer screening through implementation of office systems in community primary care practices. DESIGN: Randomized, controlled trial with primary care practices assigned to either the intervention group or control group, with the practice as the unit of analysis. SETTING: Twenty mostly rural counties in North Carolina. PARTICIPANTS: Physicians and staff of 62 randomly selected family medicine and general internal medicine practices, primarily fee-for-service, half group practices and half solo practitioners. INTERVENTION: Physician investigators and facilitators met with practice physicians and staff over a period of 12 to 18 months to provide feedback on breast cancer screening performance, and to assist these primary care practices in developing office systems tailored to increase breast cancer screening. MEASUREMENTS AND MAIN RESULTS: Physician questionnaires were obtained at baseline and follow-up to assess the presence of five indicators of an office system. Three of the five indicators of office systems increased significantly more in intervention practices than in control practices, but the mean number of indicators in intervention practices at followup was only 2.8 out of 5. Cross-sectional reviews of randomly chosen medical records of eligible women patients aged 50 years and over were done at baseline (n = 2,887) and follow-up (n = 2,874) to determine whether clinical breast examinations and mammography, were performed. Results for mammography were recorded in two ways, mention of the test in the visit note and actual report of the test in the medical record. These reviews showed an increase from 39% to 51% in mention of mammography in intervention practices, compared with an increase from 41% to 44% in control practices (p = .01). There was no significant difference, however, between the two groups in change in mammograms reported (intervention group increased from 28% to 32.7%; control group increased from 30.6% to 34.0%, p = .56). There was a nonsignificant trend (p = .06) toward a greater increase in performance of clinical breast examination in intervention versus control practices. CONCLUSIONS: A moderately intensive outreach intervention to increase rates of breast cancer screening through the development of office systems was modestly successful in increasing indicators of office systems and in documenting mention of mammography, but had little impact on actual performance of breast cancer screening. At follow-up, few practices had a complete office system for breast cancer screening. Outreach approaches to assist primary care practices implement office systems are promising but need further development. PMID- 9734787 TI - Increasing breast and cervical cancer screening in low-income women. AB - OBJECTIVE: To determine if women would have higher breast and cervical cancer screening rates if lay health advisers recommended screening and offered a convenient screening opportunity. DESIGN: Controlled trial. SETTING: Urban county teaching hospital. PARTICIPANTS: Women aged 40 years and over attending appointments in several non-primary-care outpatient clinics. INTERVENTIONS: Lay health advisers assessed the participants' breast and cervical cancer screening status and offered women in the intervention group who were due for screening an appointment with a female nurse practitioner. MEASUREMENTS AND MAIN RESULTS: Screening rates at baseline and at follow-up 1 year after the intervention were determined. At follow-up, the mammography rate was 69% in the intervention group versus 63% in the usual care group (p = .009), and the Pap smear rate was 70% in the intervention group versus 63% in the usual care group (p = .02). In women who were due for screening at baseline, the mammography rate was 60% in the intervention group versus 50% in the usual care group (p = .006), and the Pap smear rate was 63% in the intervention group versus 50% in the usual care group (p = .002). The intervention was effective across age and insurance payer strata, and was particularly effective in Native American women. CONCLUSIONS: Breast and cervical cancer screening rates were improved in women attending non-primary-care outpatient clinics by using lay health advisers and a nurse practitioner to perform screening. The effect was strongest in women in greatest need of screening. PMID- 9734788 TI - Older persons' preferences for site of treatment in acute illness. AB - OBJECTIVE: To explore how older persons form preferences for site of medical care by examining their perceptions of home care and hospital care. DESIGN: Qualitative analysis of in-depth interviews using the constant comparative method. SETTING: Respondents' homes. PARTICIPANTS: Twenty-nine persons age 65 to 89 years who had been hospitalized with congestive heart failure, chronic obstructive pulmonary disease, or pneumonia and were receiving home care services. MAIN RESULTS: Respondents, who thought of home care only as a means to provide low-intensity and low-frequency services, were initially skeptical about expanded home care services to treat acute illness. Regardless of their opinions about home and hospital, all respondents preferred the site associated with the greatest chance of survival. If the sites offered equal survival, 52% of the respondents preferred treatment at home because of freedom from the constraints of the hospital and the comfort of familiar surroundings. For respondents who preferred the hospital, the home represented a frightening and lonely place to be sick. Respondents' views of the home and hospital were shaped by their social supports, self-reliance, religious beliefs, and past illness experiences. CONCLUSIONS: Because survival appears to be the most important determinant of preference, home treatment of acute illness is a viable alternative only if it provides outcomes equivalent to those of hospitalization. Strongly held perceptions that home care can only be a low-intensity service may limit preferences for home treatment. When expected outcomes at the two sites are similar, the challenge to the health care system will be incorporating patient preference about the process of care into decisions about the appropriate site of care. PMID- 9734789 TI - Patterns and determinants of multiple provider use in patients with acute low back pain. AB - OBJECTIVE: To describe the patterns of provider use associated with an acute episode of nonspecific low back pain and their impact on cost. METHODS: The analysis is based on a prospective cohort study of patients with acute low back pain followed until they recovered completely or to 6 months. Patients were followed after an initial visit to one of four provider types: private primary care physician, chiropractor, orthopedic surgeon, or HMO primary care physician. Follow-up interviews were conducted at baseline, 2, 4, 8, 12, and 24 weeks; 1,580 (97%) of the participants completed the 6-month follow-up. MAIN RESULTS: Seventy nine percent of patients saw only the initial provider who began their care for low back pain. Logistic regression revealed that duration of pain prior to initial visit, sciatica, higher Roland disability score, days to functional recovery, interval to complete recovery, referral by initial provider, disk attribution, satisfaction, and the type of index provider were significantly (p < .05) associated with seeking care from multiple provider types. Age, race, gender, and education were not significant. The adjusted proportions of multiple provider type use were 14% (95% confidence interval [CI] 11%, 17%) for the private primary care provider stratum; 19% (95% CI 16%, 23%) for the chiropractic stratum; 30% (95% CI 23%, 37%) for the orthopedic stratum; and 9% (95% CI 5%, 14%) for the HMO primary care physician stratum. Cost of seeing only the index provider was $439 (95% CI $404, $475), and cost of seeing multiple provider types was $1,137 (95% CI $1,064, $1,211) based on the adjusted model. CONCLUSIONS: Use of multiple provider types, is associated with several factors, one of which is the initial provider type. The cost of such use is significant. PMID- 9734790 TI - Analyzing the time and value of housestaff inpatient work. AB - OBJECTIVE: To determine time allocation and the perceived value to education and patient care of the weekday activities of internal medicine housestaff on inpatient rotations and to compare the work activities of interns and residents. DESIGN: An observational study. We classified activities along five dimensions (association, location, activity, time, and value), developed a computer-assisted self-interview survey, and demonstrated its face and content validity, internal consistency, and interrater reliability. Subjects were assigned survey computers for 5 consecutive weekdays over a 24-week period, into which they entered data when prompted several times a day. SETTING: The medical service of a university affiliated Veterans Administration Medical Center. PARTICIPANTS: Sixty housestaff (36 interns, 24 residents) rotating on the inpatient wards. MEASUREMENTS AND MAIN RESULTS: We analyzed activities according to content (direct patient care, indirect patient care, education), association, and location. Likert-scale ratings of perceived value to education and patient care were also obtained. Housestaff provided complete responses to 3,812 (95%) of 3,992 prompts by a median of 11 seconds; 93% of responses were logically consistent across the measured dimensions. Housestaff spent more time in indirect patient care (56%) than in direct patient care (14%) or educational activities (45%). Formal educational activities had the highest educational value (66 on 0-100 scale), and direct care had the highest value to patient care (81). Over 30% of time was spent in administrative activities, which had low educational value(40). Compared with residents, interns allocated significantly less time to educational activities (38% vs 57%) and more time to lower-value activities such as documentation (19% vs 12%). CONCLUSIONS: Improved data collection methods demonstrate that housestaff in our program, particularly interns, spend much of their workday in activities that are low in educational and patient care value. Selective elimination or delegation of such activities would preserve higher value experiences during reductions in overall inpatient training time. Planners can use automated random sampling to guide the rational redesign of housestaff work. PMID- 9734792 TI - Inside "Pandora's box": abused women's experiences with clinicians and health services. AB - OBJECTIVE: To explore the attitudes and experiences of abused women to identify characteristics that helped or hindered abuse disclosure to clinicians and to determine how women viewed potential interventions to improve detection and treatment in a medical setting. DESIGN: Focus group data conducted and analyzed with qualitative methodology. SETTING: Three community-based mental health centers and one women's shelter. PARTICIPANTS: Twenty-one women in group therapy for domestic violence. MAIN RESULTS: Eighteen (86%) of the 21 women had seen their "regular doctor" in the prior year; only 1 in 3 had discussed the abuse with the clinician. The major discussion themes were medical problems that were exacerbated with abuse, lack of ability to access medical care due to abuser interference, emotional attitudes about abuse that acted as barriers to disclosure, clinician characteristics that helped or hindered disclosure, and treatment experiences and preferences. Women described how their medical problems began or worsened during the abusive period. one in three women described how abusers blocked them from receiving medical care. Women reported intense shame about the abuse and described their self-denial of abuse. Women stated they were inclined to discuss abuse if they felt the clinician was perceived to be caring, was easy to talk to, had a protective manner, or if the clinician offered a follow-up visit. There was no consistent clinician gender preference among the women. One in four women had received psychotropic medication for problems associated with abuse. Many feared addiction, or a loss of alertness, increasing their risk for more abuse. CONCLUSIONS: Many abused women experience worsening health and seek medical care; most do not volunteer a history of violence even to their regular clinicians. Many of the barriers to disclosure of abuse could be overcome by a physician's knowledge of the link between abuse and medical illness, an understanding of the women's emotions about abuse, and her treatment preferences. PMID- 9734793 TI - Aggression and violence directed toward physicians. PMID- 9734791 TI - Factors associated with antibiotic use for acute bronchitis. AB - OBJECTIVES: To describe the clinical features of adults diagnosed with acute bronchitis, and to identify clinical variables associated with antibiotic treatment of acute bronchitis. DESIGN: Prospective, cohort study. SETTING: Primary care office practices at a group-model HMO in the Denver metropolitan area. PATIENTS/PARTICIPANTS: Patients were adults seeking care for acute respiratory illnesses. Participating clinicians included internists, family medicine physicians, nurse practitioners, physician assistants, and registered nurses. MEASUREMENTS AND MAIN RESULTS: Clinicians voluntarily completed encounter forms for patients presenting with acute respiratory illnesses between February and May, 1996. Acute bronchitis was the primary diagnosis in 16% of acute respiratory illness visits (n = 1,525). The most frequent symptoms of acute bronchitis were cough (92%), phlegm production (63%), "runny nose" (50%), and throat pain (50%). The most frequent physical examination findings were pharyngeal erythema (45%), cervical lymphadenopathy (19%), wheezes (18%), and rhonchi (17%). Antibiotics were prescribed to 85% of patients diagnosed with acute bronchitis. Purulent nasal discharge by patient report, and sinus tenderness on physical examination were moderately associated with antibiotic treatment (p = .06 and .08, respectively). Antibiotic prescription rates did not vary by patient age or gender, duration of illness, days of work lost due to illness, or clinician type. CONCLUSIONS: Acute bronchitis is frequently treated with antibiotics in ambulatory practice. The clinical factors we identified to be associated with antibiotic use for acute bronchitis appear to play a minor role in explaining the excessive use of antibiotics for this condition. These findings suggest that clinicians use the diagnosis of acute bronchitis as an indication for antibiotic treatment, despite clinical trials and expert recommendations to the contrary. PMID- 9734794 TI - Teaching residents about complementary and alternative medicine in the United States. PMID- 9734795 TI - Patients discharged against medical advice from a general medicine service. AB - This study compares the demographic features and hospital course of all 472 patients discharged against medical advice from the general medicine service of an urban teaching hospital between 1984 and 1995 and 1,113 control patients discharged with physician approval. In the multivariate analysis, younger age (odds ratio [OR] 0.97 per year; 95% confidence interval [CI] 0.96, 0.98), male gender (OR 1.9; 95% CI 1.4, 2.4), lack of health insurance (OR 2.0; 95% CI 1.3, 3.1), Medicaid applicant or recipient status (OR 2.2; 95% CI 1.6, 3.1), admission through the emergency department (OR 2.2; 95% CI 1.4, 3.5), and lack of a personal attending physician at the time of admission (OR 2.1; 95% CI 1.6, 2.8) increased the odds of discharge against medical advice. Fifty-four percent of patients who left against medical advice were readmitted to the hospital during the study period; 98% were then discharged with physician approval. Patients who left the hospital against medical advice included many disadvantaged individuals without ongoing primary care. PMID- 9734796 TI - Targeting depression interviewing. PMID- 9734797 TI - Our tax dollars at work. PMID- 9734798 TI - MEDLINE World Wide Web sites. PMID- 9734799 TI - On-eye power characteristics of soft contact lenses. PMID- 9734800 TI - Retinitis pigmentosa inversa. AB - BACKGROUND: Retinitis pigmentosa (RP) is one of the most common inherited retinal diseases, with a prevalence of about 1 in 3500 to 4500. Retinitis pigmentosa inversa is a rare variant of this disorder characterized by areas of choroidal degeneration with pigment migration and bony spicule formation in the macular area. In contrast to more typical forms of RP, this anomaly destroys central vision, leaving peripheral vision intact. CASE REPORT: A 47-year-old white male was followed for about 7 years with evidence of progressive retinal pigment epithelial atrophy and hyperpigmentation affecting both maculae. Since 1970, he had noted difficulty seeing at night as well as an acquired hearing deficit that appeared to be getting worse, ultimately impairing his ability to safely drive a truck. Medical history was positive for either chloroquine or hydroxychloroquine use for 2 to 3 years as malaria prophylaxis while he served in Vietnam. In addition, his father in Louisiana had visual loss of unknown cause. During the 7 year period, the condition progressed rapidly. The patient became virtually blind secondary to visual acuity loss with dense central and paracentral scotomas. The peripheral visual fields remained intact. After several years of extensive examinations, including laboratory, electroretinography, and genetic testing, a definitive diagnosis of RP inversa was made. DISCUSSION: RP inversa is a rare form of tapetoretinal degeneration that is characterized by decreased central vision with normal peripheral vision. A recessive form of inheritance has been postulated but never substantiated. Although there is currently no treatment, recent studies have indicated that 15,000 IU of vitamin A palmitate daily may slow the progression of retinitis pigmentosa; however, it is unknown whether this treatment would be effective for the inverse form of RP. Differential diagnoses include Leber's congenital amaurosis, central gyrate atrophy, central areolar choroidal sclerosis, progressive cone-rod dystrophy, syphilitic retinopathy, retinal toxicity from phenothiazine use, and chloroquine/hydroxychloroquine retinopathy. PMID- 9734801 TI - Vision screening of preschool children: evaluating the past, looking toward the future. AB - Vision problems of preschool children are detectable with a comprehensive eye examination; however, it is estimated that only 14% of children below the age of 6 years receive an eye examination. Screening is advocated as a cost-effective alternative to identify children in need of further vision care. Thirty-four states recommend or require vision screening of preschool children. Although laws and guidelines exist, only 21% of preschool children are actually screened for vision problems. There is little agreement concerning the best screening methods, and no validated, highly effective model for screening vision of preschool children. Newer screening tests have been designed specifically for preschool populations, and can be administered by lay screeners. Many have not been validated. Several are recommended by states or organizations without convincing scientific evidence of their effectiveness. This paper summarizes current laws and guidelines for preschool vision screening in the United States, reviews advantages and disadvantages of several test procedures, and provides recommendations for developing future preschool vision screening programs. PMID- 9734802 TI - Assessment of high and low contrast visual acuity after photorefractive keratectomy for myopia. AB - BACKGROUND: Our aim was to assess visual acuity using standardized charts and illumination conditions after photorefractive keratectomy. METHODS: High and low contrast visual acuity were measured on Bailey-Lovie logarithm of the minimum angle of resolution (LogMAR) charts under high and low illumination conditions on 105 photorefractive keratectomy patients who had been treated with the Summit (N = 60) or the VISX (N = 45) excimer laser. RESULTS: Best corrected visual acuity was reduced in the treated eye compared with the untreated control eye under all test conditions, with the greatest differences under conditions of low contrast and low illumination. Reduction of acuity under low contrast and low illumination was related to small optic zone sizes and steep ablation edge profiles found in Summit-treated eyes. In the VISX-treated eyes, high contrast acuity was reduced in the presence of central topographical irregularities, subepithelial haze, and higher myopic corrections. CONCLUSIONS: Testing conditions such as those described here may be useful in quantifying vision degradation in suboptimal viewing conditions and among patients with vague complaints. PMID- 9734803 TI - Pilot study on the use of impression cytology specimens for quantitative assessment of the surface area of bulbar conjunctival cells. AB - PURPOSE: To assess whether impression cytology samples could be used to assess cell conjunctival cell surface areas, and to obtain an estimate of these for normal appearing cells vs. those with squamous metaplasia. METHODS: Small sheets of nasal bulbar conjunctival cells (c. 0.03 mm2) were obtained from 17 subjects aged 25 to 78 years by manual application of a 0.45 microm Millipore filter without topical anesthetic. Outlines of cells, visible as a rarefaction of the general cell cytoplasmic staining, were made on an optical overlay and then planimetry was carried out. RESULTS: Reasonable estimates (with +/-2.5%) of the average cell area can be obtained from samples containing contiguous sets of 100 cells. From 14 separate samples of normal cells [subjective assignment of nucleus:cytoplasm (N:C) ratios of 1:1 to 1:2], cell surface area values ranged from 11 to 426 microm2, with an overall average value of 108 +/- 43 microm2. Samples from individuals with symptoms suggestive of borderline dry eye tended to have slightly higher proportions of larger cells (so increasing the average cell area), and there was a trend for samples from older individuals to contain more smaller cells (so decreasing the average cell area). In marked contrast, cells showing evidence of squamous metaplasia and with N:C ratios of 1:6 or higher were found to have an average area of 543 +/- 76 microm2 (range 310 to 1529 microm2). CONCLUSIONS: Morphometry of impression cytology specimens is possible. The average cell size in normal individuals is considerably smaller than previously reported (108 vs. 585 microm2), but this is consistent with a range of studies on animal tissue using scanning electron microscopy. PMID- 9734804 TI - The repeatability of tear mucus ferning grading. AB - PURPOSE: Rolando's classification system for tear mucus ferning patterns subjectively assigns grades based on the size and spacing between ferns. Grade 1 and 2 patterns are considered "normal," whereas grade 3 and 4 patterns are often associated with keratoconjunctivitis sicca. This study was designed to examine the intraobserver and interobserver repeatability of Rolando's system. METHODS: Photographic slides (N = 418) of portions of tear ferning patterns were randomly assembled, numbered, and graded by two investigators (I1 and I2). I1 graded the slides twice; the slides were remixed under masked conditions between the first and second runs. I2 graded the slides once, independent of I1. RESULTS: For the four-grade system, intraobserver agreement was 85.41% (simple kappa = 0.75; 95% confidence interval ]CI] = 0.69-0.82). Interobserver agreement was 80.62% (kappa = 0.67; CI = 0.60-0.74) and 86.12% (kappa = 0.75; CI = 0.68-0.83) for the first and second runs of I1, respectively. Analysis of the ability to simply classify normal (grade 1 and 2) from abnormal (grade 3 and 4) patterns revealed intraobserver repeatability of 94.50% (kappa = 0.76; CI = 0.67-0.86). Interobserver agreement was 92.10% (kappa = 0.65; CI = 0.56-0.75) and 94.26% (kappa = 0.71; CI = 0.62-0.81) for the first and second runs, respectively. CONCLUSION: Based on the high rate of agreement between intraobserver and interobserver trials, Rolando's grading system appears to be an easy and consistent method for the classification of tear ferning patterns. PMID- 9734805 TI - Development of phoria in children. AB - BACKGROUND: Although the prevalence of phoria in adults is well documented, data are scarce on phoria in children. We present results using modified clinical technique vision screening and data from the Orinda Longitudinal Study of Myopia on a large, population-based sample of nonstrabismic children to document the prevalence of phoria with age. METHODS: We collected cross-sectional (N = 1495) and longitudinal (N = 469) data. Phoria data were collected by cover tests administered by one observer who subjectively classified phoria as orthophoria, esophoria, or exophoria in 2 delta steps. RESULTS: Ninety-seven percent of the children were orthophoric at distance, and there were no significant changes with age. Near phoria showed a more normal distribution, with a 10.8% decrease in the prevalence of exophoria (from 31.8 to 21.0%) and a 5.5% increase in the prevalence of esophoria (from 6.7 to 12.2%) between kindergarten and fifth grade. CONCLUSIONS: Children are typically orthophoric or exophoric at near, but convergent shifts occur with age. PMID- 9734806 TI - Accommodation and pupillary response in early-onset myopia among schoolchildren. AB - PURPOSE: To investigate changes in accommodation and pupillary reaction in early onset myopia among children. METHODS: An objective infrared optometer and a pupillometer were used to evaluate simultaneously tonic accommodation, accommodative response, accommodative adaptation, and pupil diameter in 19 schoolchildren with early-onset myopia and 15 age-matched emmetropic controls. RESULTS: Tonic accommodation in the early-onset myopes (1.03 D) was significantly less than in the emmetropes (1.37 D) (p < 0.05), whereas accommodative response and accommodative adaptation showed no significant difference between the early onset myopes and emmetropes. The average pupil diameter after 20 min of dark adaptation in the early-onset myopes was 4.52 mm, which was significantly smaller than that in the emmetropes (5.21 mm) (p < 0.05). During nearwork stimulation, the average pupil diameter further decreased to 3.83 mm in the early-onset myopes and 4.78 mm in the emmetropes (p = 0.003). CONCLUSION: These results suggest that low tonic accommodation and small pupil diameter may play important roles in the pathogenesis of early-onset myopia among schoolchildren. PMID- 9734807 TI - The repeatability of automated and clinician refraction. AB - PURPOSE: Auto-refractors are used as a starting point for clinicians' refractions and in studies of refractive error. We investigated the repeatability of the Hoya AR-570 and clinician refraction. METHODS: Eighty-six subjects, aged 11 to 60 years, were recruited by mailing inquiries to 500 randomly selected patients who had received recent examinations at the University of California Optometric Eye Center. Contact lens wearers, patients with best corrected visual acuity worse than 20/30 in either eye, and patients with a history of diabetes were excluded. Each subject was examined by two clinicians during one visit. The first clinician obtained five auto-refractor readings for each eye (which were later averaged), performed a balanced subjective refraction (with spherical masking lenses in the phoropter), and repeated the automated refractor measurements. This protocol was then repeated by the second clinician. Clinicians were randomized with regard to testing order and masked to automated refractor results, each other's refractions, and previous spectacle prescriptions. RESULTS: To quantify repeatability, we used mixed model analyses of variance to estimate the appropriate variance components while accounting for the correlation among, for example, repeated measurements of the same eye. Astigmatic data were analyzed by converting into Fourier form: two cross-cylinders at axis 0 degrees (J0) and axis 45 degrees (J45). For mean spherical equivalent, the average difference between five averaged automated refractor readings, taken by two different optometrists, was +0.02 D (95% limits of agreement = -0.36 to +0.40 D). The average difference between the two optometrists' subjective refractions was -0.12 D (95% limits of agreement = -0.90 to +0.65 D). The 95% limits of agreement for the automated refractor were about half those of the clinician for both astigmatic terms (J0 and J45) and for all comparisons. CONCLUSIONS: Automated refraction is more repeatable than subjective refraction and therefore more appropriate for studies of myopia progression. PMID- 9734808 TI - A modified card procedure for measuring human infant color vision. AB - To improve test efficiency, we modified our previously introduced contrast/color card test by including a patterned test stimulus and reducing the number of stimuli in both experimental phases. Compared with the prototype, completion rate improved substantially (79 vs. 37%) but test time decreased only modestly (19 vs. 21 min). Achromatic contrast discrimination improved threefold (mean, 0.06 vs. 0.20 log units), but the percentage of 2-month-old infants who discriminated (from gray) 660-nm red (86 vs. 80%) and 580-nm yellow (52 vs. 55%) was consistent. In addition, 48% discriminated 574-nm yellow-green. Moreover, because 88% of infants' failures included the respective adult brightness/luminance match, a small range of relative luminances is adequate for testing infant color vision. PMID- 9734809 TI - The polyphosphate kinase gene of Pseudomonas aeruginosa. AB - We have cloned and sequenced a gene encoding polyphosphate kinase (PPK) from Pseudomonas aeruginosa PAO1. The gene immediately follows the hemB gene encoding porphobilinogen synthase responsible for heme synthesis. The predicted amino acid sequence of P. aeruginosa PPK is similar to those of PPKs previously characterized except that it possesses an extra stretch of 46 amino acids at its N-terminus, which has significant similarity to the Ras-related protein ARA5 of Arabidopsis thaliana. When P. aeruginosa PPK was overproduced in Escherichia coli, ATP-dependent polyphosphate-synthesizing activity was drastically enhanced, confirming that the protein is a PPK. PMID- 9734810 TI - A physical map of Arabidopsis thaliana chromosome 3 represented by two contigs of CIC YAC, P1, TAC and BAC clones. AB - We have constructed a physical map of Arabidopsis thaliana chromosome 3 by ordering the clones from CIC YAC, P1, TAC and BAC libraries using the sequences of a variety of genetic and EST markers and terminal sequences of clones. The markers used were 112 DNA markers, 145 YAC end sequences, and 156 end sequences of P1, TAC and BAC clones. The entire genome of chromosome 3, except for the centromeric and telomeric regions, was covered by two large contigs, 13.6 Mb and 9.2 Mb long. This physical map will facilitate map-based cloning experiments as well as genome sequencing of chromosome 3. The map and end sequence information are available on the KAOS (Kazusa Arabidopsis data Opening Site) web site at http://www.kazusa.or.jp/arabi/. PMID- 9734811 TI - Prediction of the coding sequences of unidentified human genes. X. The complete sequences of 100 new cDNA clones from brain which can code for large proteins in vitro. AB - As an extension of our cDNA analysis for deducing the coding sequences of unidentified human genes, we have newly determined the sequences of 100 cDNA clones from a set of size-fractionated human brain cDNA libraries, and predicted the coding sequences of the corresponding genes, named KIAA0611 to KIAA0710. In vitro transcription-coupled translation assay was applied as the first screening to select cDNA clones which produce proteins with apparent molecular mass of 50 kDa and over. One hundred unidentified cDNA clones thus selected were then subjected to sequencing of entire inserts. The average size of the inserts and corresponding open reading frames was 4.9 kb and 2.8 kb (922 amino acid residues), respectively. Computer search of the sequences against the public databases indicated that predicted coding sequences of 87 genes were similar to those of known genes, 62% of which (54 genes) were categorized as proteins related to cell signaling/communication, cell structure/motility and nucleic acid management. The expression profiles in 10 human tissues of all the clones characterized in this study were examined by reverse transcription-coupled polymerase chain reaction and the chromosomal locations of the clones were determined by using human-rodent hybrid panels. PMID- 9734813 TI - Transmembrane-domain trapping: a novel method for isolation of cDNAs encoding putative membrane proteins. AB - We have developed a method that enables us to isolate cDNAs of putative membrane proteins. The system is designed to isolate a cDNA which can provide the transmembrane domain to the extracellular part of the IL-2 receptor alpha chain. We constructed a p18Mac vector by putting part of the IL-2 receptor alpha chain cDNA that encoded its signal sequence and extracellular domain, a cDNA cloning site and a poly(A) additional signal after a strong promoter SRalpha. If a cloned cDNA provides a transmembrane domain in-frame, the extracellular domain of the IL 2 receptor alpha chain will be expressed on the surface of the transfected cells. Otherwise, the chimeric protein will be either secreted or retained inside the transfected cells. We made a cDNA library using p18Mac and screened for cDNA clones which allowed the expression of the extracellular domain of the IL-2 receptor alpha chain on the cell surface. Of the 2000 clones screened, 5 clones were scored as positive. Partial sequence analysis revealed that one clone encoded the amyloid precursor protein, two others encoded mitochondrial proteins and the rest were new. These results suggest the system is effective in isolating cDNAs encoding putative membrane proteins. PMID- 9734812 TI - The primary structure and genomic organization of five novel transcripts located close to the Huntington's disease gene on human chromosome 4p16.3. AB - Five distinct novel transcripts (RES4-22, -23, -24, -25 and -26) that mapped to the 1-Mb interval between D4S180 and D4S183 on human chromosome 4p16.3 close to the Huntington's disease (HD) gene were isolated, and the structure and exon/intron organization of each gene were thoroughly analyzed. The transcripts of the RES4-22, -23 and -24 genes each have several isoforms by alternative splicing and these have also been defined. Two transcripts, RES4-24 and RES4-25, reside in the same genomic region with opposite polarities and they also clearly overlap. Among these transcripts, RES4-26 was found to encode a novel zinc finger protein. The transcript map based upon our current level of analysis combined with data from previous studies reveals the gene-rich nature and the intricate organization of the genes in the HD locus. PMID- 9734814 TI - Sequence analysis of the Bacillus subtilis 168 chromosome region between the sspC and odhA loci (184 degrees-180 degrees). AB - The nucleotide sequence of 45,389 bp in the 184 degrees-180 degrees region of the Bacillus subtilis chromosome, containing the cge cluster, which is controlled by the sporulation regulatory protein GerE, was determined. Fifty-four putative ORFs with putative ribosome-binding sites were recognized. Seven of them correspond to previously characterized genes: cgeB, cgeA, cgeC, cgeD, cgeE, ctpA, and odhA. The deduced products of 25 ORFs were found to display significant similarities to proteins in the data banks. We have identified genes involved in detoxification, cell walls, and in the metabolism of biotins, purines, fatty acids, carbohydrates and amino acids. The remaining 22 ORFs showed no similarity to known proteins. Both an attachment site of the SPbeta prophage and 2 new putative DNA replication terminators were identified in this region. PMID- 9734815 TI - Structural analysis of Arabidopsis thaliana chromosome 5. VI. Sequence features of the regions of 1,367,185 bp covered by 19 physically assigned P1 and TAC clones. AB - Nineteen P1 and TAC clones, which have been mapped on the fine physical map of the Arabidopsis thaliana chromosome 5, were sequenced according to the shotgun based strategy, and their structural features were analysed. The total length of the regions sequenced in this study was 1,367,185 bp. Combining this with the regions covered by 90 P1 and TAC clones previously reported, the total length of chromosome 5 sequenced to date becomes 8,058,855 bp. On the basis of similarity search against protein and EST databases and gene modeling with computer programs, a total of 330 potential protein-coding regions were identified, bringing an average density of the genes to approximately one gene per 4.1 kb. Introns were identified in 81.0% of the potential protein genes for which the entire gene structure was predicted, with an average number per gene of 4.2 and an average length of the introns of 180 bp. The RNA-coding genes identified were 9 tRNA genes corresponding to 8 amino acid species and 2 genes for U2 nuclear RNA. These sequence features are essentially identical to those in the previously reported sequences. The sequence data and gene information are available on the World Wide Web database KAOS (Kazusa Arabidopsis data Opening Site) at http://www.kazusa.or.jp/arabi/. PMID- 9734816 TI - A simple, two-color fluorescence detection method for membrane blotting analysis using alkaline phosphatase and horseradish peroxidase. AB - We have developed a one-step, two-color fluorescence detection method using simultaneously two fluorogenic substrates for both Southern and Western blots on nylon membranes. For this enzyme-mediated reporter system, a mixture of (i) 3 hydroxy-N-2'-biphenyl-2-naphthalenecarboxamide phosphate ester (HNPP), a substrate for alkaline phosphatase and (ii) N-(4-amino-5-methoxy-2 methylphenyl)benzamide (AMMB), a fluorogenic substrate for horseradish peroxidase was used. The reaction with these substrates produces blue (HNPP) and yellow (AMMB) fluorescent signals under ultraviolet light (302 nm). Therefore, this simple method allows the simultaneous visualization of two different targets on a single nylon membrane, e.g. nucleic acids or proteins. PMID- 9734817 TI - pH, cholesterol sulfate, and fatty acids affect the stratum corneum lipid organization. AB - Lipid mixtures prepared from cholesterol (CHOL), isolated ceramides (CER), and free fatty acids can serve as attractive tools to study the role various stratum corneum (SC) lipids or microenvironmental conditions play in the SC lipid organization, as the phase behavior in these mixtures and in SC are similar: two lamellar phases with periodicities of approximately 6 and 13 nm are present. Because pH and cholesterol sulfate (CSO4) gradients exist in SC and may affect the local SC lipid organization, the effects of pH and CSO4 on lipid phase behavior was examined. X-ray diffraction studies with CHOL:CER mixtures revealed that the lamellar ordering at pH 5 and 7.4 were similar: both the short and the long periodicity phases were present. Upon addition of free fatty acids the phase behavior became pH dependent; the long periodicity phase being more dominant at pH 7.4 than at pH 5. Similar observations have been made upon addition of CSO4. Furthermore, only in the presence of CSO4 did phase-separated CHOL disappear, indicating that CHOL completely dissolves in the lamellar phases. A major phase change from an hexagonal to an orthorhombic lateral packing has been observed in the presence of free fatty acids. Furthermore, in the presence of CSO4 next to orthorhombic also liquid lateral packing could be detected. In contrast to lamellar ordering, changes in pH did not affect the lateral packing in any of the lipid mixtures studied. PMID- 9734818 TI - Ontogeny of the epidermal permeability barrier. AB - A competent permeability barrier must be present by the end of gestation to allow for life in a terrestrial environment. Indeed, early preterm infants display serious complications of skin immaturity. Yet, regardless of their degree of prematurity, all infants quickly develop a competent barrier. To learn more about the mechanisms and regulation of barrier ontogeny, we have utilized late gestation fetal rodents. In 19-21 d fetal rats, we showed that barrier competence is accompanied by both enhanced epidermal development and formation of extracellular lamellar membranes in the stratum corneum. The identical sequence and time-course occurs when fetal rat skin is cultured in a serum-free medium. Glucocorticoids, thyroid hormone (T3), and estrogen accelerate, while androgens delay barrier formation both in utero and in the in vitro system, explaining the poorer outcome of premature males versus females. But neither T3 nor glucocorticoids are absolutely required for barrier development. Lifting fetal skin cultures to an air-medium interface also accelerates barrier formation, explaining the rapid emergence of barrier competence in very premature infants. PPARalpha and FXR activators, which, like T3, heterodimerize with the nuclear receptor, RXR, also accelerate barrier development in vitro. Finally, not only the nuclear receptor family, but also Ca++ could regulate key events late in barrier development. PMID- 9734819 TI - Lamellar granule biogenesis: a role for ceramide glucosyltransferase, lysosomal enzyme transport, and the Golgi. AB - Although lamellar granules are critical to the formation of the epidermal permeability barrier and are a known marker of late keratinocyte differentiation, very little is known about the physiologic regulators of lamellar granule assembly and extrusion. Ceramide glucosyltransferase (CGT), the enzyme responsible for the synthesis of lamellar granule glucosylceramides (GlcCer; the precursors of the stratum corneum ceramides), is localized to the Golgi apparatus in other cell types. We have found that CGT is induced during keratinocyte culture differentiation coincident with increased GlcCer content and the appearance of lamellar granules. In this study we show that the differentiation related CGT induction is likely mediated at the transcriptional level. In addition, all-trans retinoic acid, a well-known inhibitor of keratinocyte differentiation, prevents the appearance of lamellar granules and decreases culture CGT activity and GlcCer content without affecting sphingomyelin or total lipid content, indicating a specific inhibition of this enzymatic pathway. These data show a direct relationship between CGT activity and epidermal differentiation, suggesting that regulation of CGT expression is a critical part of epidermal barrier generation. The differentiation dependence of CGT activity, the key role of this Golgi-localized enzyme in epidermal GlcCer synthesis, and our previous finding that ceramides are converted to GlcCer in the Golgi apparatus in keratinocyte cultures, strongly suggest a Golgi origin for lamellar granules. In contrast to CGT, the activity of the lysosomal enzymes acid lipase and glucocerebrosidase is less clearly related to epidermal differentiation and the appearance of lamellar granules, although both enzymes show striking colocalization and enrichment in a subcellular lamellar granule fraction derived from pig epidermis. Acid lipase activity in the lamellar granule fraction was found to contain primarily a small lysosomal form of the enzyme, whereas total acid lipase secreted by keratinocyte cultures was found to contain a mannose-6 phosphorylated large prelysosomal form as well as a small lysosomal form. That secreted acid lipase activity is derived from both prelysosomal and lysosomal compartments suggests there may be multiple pathways by which lysosomal enzymes are secreted from keratinocytes. The combined secretion of lipid and lysosomal enzymes from lamellar granules places these organelles in the category of "dual function" specialized secretory vesicles described in certain other cell types. Electron microscopic images of lamellar granules show shapes consistent with cross-sections of tubules or buds from tubules in addition to vesicles. These images provide evidence for the involvement of trans-Golgi network tubules and/or buds in lamellar granule synthesis and secretion. PMID- 9734820 TI - The secretory granular cell: the outermost granular cell as a specialized secretory cell. AB - The contents of epidermal lamellar bodies (LB) are delivered selectively to the intercellular spaces at the stratum granulosum (SG)-stratum corneum (SC) interface. We assessed the subcellular basis for LB secretion first by confocal microscopy, following labeling with Nile red or NBD-ceramide, which reveals a tubulo-reticular membrane system within the apical cytosol of the outermost SG cell layer under basal conditions, changing to a more peripheral staining pattern when secretion is stimulated. Ultrastructural study demonstrates that this network is composed of a widely disbursed trans-Golgi-like network (TGN), associated with arrays of contiguous LB, and deep invaginations of the SG-SC interface. Under basal conditions, limited fusion of apically directed LB leads to deep, interconnected invaginations of the apical plasma membrane, resulting in the formation of an extensive, honeycomb extension of the SG-SC interface. Still deeper invaginations and more extensive organelle fusion develop after the epidermis is acutely permeabilized by either acetone treatment, sonophoresis, or iontophoresis. Finally, nascent LB appear to bud off cisternae of the TGN, a process that appears to accelerate after barrier disruption. The deep invaginations of the SG-SC interface; the wide distribution of the TGN within the apical cytosol; the association of nascent LB with the TGN; and the rapid fusion of LB with these invaginations, deep within the cytosol, account for (i) the polarized secretion of LB from the apex of the outermost SG cell, and (ii) the rapid LB-secretory response to barrier perturbations. Finally, our results point to the outermost SG cell as a uniquely specialized secretory cell. We propose the term "secretory granulocyte" to encompass the specialized features of these cells. PMID- 9734821 TI - What's water got to do with it? A nuclear magnetic resonance study of molecular motion in pig stratum corneum. AB - The 1H and 2H magnetic resonance signals from pig stratum corneum were measured as a function of hydration. The 1H free induction decay contained two components, one motionally restricted and the other isotropically mobile on the timescale of 10(-5) s. From its T2 decay, the mobile signal was further subdivided into two components; one, which was 11% of the signal and 5.5% of the mass of the dehydrated stratum corneum, was assigned to nonaqueous tissue (likely hydrocarbons) and the other to water. As water content increased from 0 to 0.25 gH2O/gSC, the second moment of the motionally restricted signal decreased from 5.4 x 10(9) to 3.6 x 10(9) s(-2), whereas the water T2 time increased from less than 0.3 ms to 3.3 ms. The 2H quadrupolar echo from stratum corneum hydrated in 2H2O had a signal from motionally restricted deuterons, attributed to deuterons exchanged onto O-H and N-H groups, and a mobile signal from 2H2O. The amount of exchange, 9.5% of the hydrogen sites in the motionally restricted fraction, was close to the number of exchangeable sites on keratin, the most abundant protein in stratum corneum. Our results are consistent with a model in which the bulk of the water interacts closely with the corneocytes in stratum corneum. PMID- 9734822 TI - Characterization of the stratum corneum lipid matrix using fluorescence spectroscopy. AB - Using fluorescence techniques, we studied the dynamics of the lipid bilayer matrix of human stratum corneum (SC) and compared the results with that of distearoyl-phosphatidylcholine (DSPC). We employed a series of 9-anthroyloxy fatty acids (AF) that partitioned into the bilayer, enabling us to evaluate this structure as a function of depth within the lamellae. With AF probes, the re orientation of the fluorophore is known to be affected by the polarity, hydrogen bonding, and rigidity of the surrounding medium, altering the emission maximum and lifetime in the excited state. In addition, we evaluated quenching, in which iodide collides with the fluorophore, revealing information on the accessibility of the fluorophore located in the bilayer. The emission and lifetime data showed that the reorientation of the fluorophore in SC was more hindered than in DSPC, indicating that SC bilayers were more rigid than DSPC bilayers. Quenching data of both SC and DSPC indicated that the deeper the fluorophore was positioned in the bilayer, the less accessible it was to iodide, pointing to a gradient in accessibility. In addition, the quenching results also showed that the SC is less accessible to iodide than in DSPC. The observed differences in bilayer rigidity and quencher accessibility between the two systems can be explained by differences in lipid composition and hydration. Whereas the DSPC bilayer consists of phospholipids, SC bilayers are composed of more anhydrous lipids like cholesterol and ceramides, which form a tight bilayer packing. In this way SC lipids exist in a relatively anhydrous and rigid environment, forming an effective diffusion barrier to water and ions. PMID- 9734823 TI - Determination of the pH gradient across the stratum corneum. AB - The objective of this work was to determine the pH gradient profile across hairless mouse stratum corneum (SC) using ratiometric laser scanning confocal microscopy imaging and direct pH measurement with a flat surface electrode. Dual emission ratiometric imaging used the fluorophore, carboxy seminaphthorhodafluor 1, which displays a pH-dependent shift in its emission spectrum. The assay developed was unsuccessful, however, because (i) the pKa of seminaphthorhodafluor (approximately 7.5) makes it insufficiently sensitive to the normal pH gradient (4-7.4) across mammalian SC, and (ii) the unexpectedly high buffering capacity of the skin precluded a meaningful calibration of the system. In the second method, pH measurements with a flat surface electrode were recorded as the hairless mouse SC was progressively tape stripped. In the superficial SC, the pH remained relatively constant (approximately 5.9); further removal of the barrier resulted in a steady increase in pH to approximately 7. In conclusion, improved methodology is clearly required to characterize precisely the pH profile across the SC. PMID- 9734824 TI - Stratum corneum lipid composition and structure in cultured skin substitutes is restored to normal after grafting onto athymic mice. AB - Restoration of an epidermal barrier is a definitive requirement for wound closure. Cultured skin substitutes grafted onto athymic nude mice were used as a model for a long-term study of stratum corneum barrier lipid metabolism and organization. Samples of stratum corneum collected after 12 and 21 d in vitro and 6, 11, and 24 mo postgrafting were examined for their lipid and fatty acid composition, and their lipid organization and structure using electron microscopy and small angle X-ray diffraction, respectively. All of these methods confirm the impaired barrier function of cultured skin substitutes in vitro, as judged from the deviations in lipid composition and from poor organization of the stratum corneum lipids that show no lamellar structure. At 6 mo postgrafting, the total stratum corneum lipid profiles of the epidermal grafts is close to that of the human stratum corneum with the exception of the presence of mouse specific lipids. The increase of ceramides 4-7 in cultured skin substitutes after grafting indicates restored activity of processes involved in the hydroxylation of fatty acids and sphingoid bases. Conversely, the ceramide profile still reveals some abnormalities (elevated content of ceramide 2 and slightly lower content of ceramide 3) and the content of long-chain fatty acids remains below its physiologic level at 6 mo postgrafting, but normalizes by 2 y postgrafting. The ultramicroscopic observations revealed the formation of lamellar extracellular lipid domains by 4 mo postgrafting. Despite these findings, the X-ray diffraction showed differences in the diffraction pattern at 2 y after grafting, suggesting that the organization of stratum corneum lipids in all epidermal grafts differs from that of the native skin. PMID- 9734825 TI - Characterization of detergent-induced barrier alterations -- effect of barrier cream on irritation. AB - To gain a better understanding of the interaction of the model detergent sodium lauryl sulfate (SLS) with the stratum corneum, we investigated systematically the ultrastructural changes of the epidermal barrier and the nucleated parts of the epidermis after the occluded application of different concentrations of SLS in human. Different application models were investigated. Two of the three irritation procedures (long duration exposure and the repetitive exposure for 3 d) provoked damage of the nucleated parts of the epidermis and alterations of the lower parts of the stratum corneum. Here, the extrusion and transformation of lamellar body derived lipids into lamellar lipid bilayers were disturbed; however, the upper portions of stratum corneum displayed intact intercellular lipid layers that contradict the long-standing belief that surfactants damage the skin by delipidization. Furthermore, we investigated ultrastructurally and by measurement of transepidermal water loss the influence and protective capacity of a lipophilic barrier cream on acute irritant contact dermatitis. The irritant contact dermatitis was induced by the standardized cumulative short application model with two SLS concentrations (0.5% and 0.75%). The cumulative type of exposure simulates daily living more realistically. Because most of the previous tests have been performed on the human forearm or back, we analyzed whether the pattern of response was similar on both sites. The back showed a higher level of irritant reaction, but the pattern of irritant response proved to be similar to the forearm. Application of the barrier cream before and during irritation showed a decrease of transepidermal water loss enhancement with 0.5% SLS by 58% (back) and 49% (arm) and after irritation with 0.75% SLS by 56% (back) and 43% (arm). Because the experimental result correlated with the clinical experience, the development of the cumulative short exposure model might help to predict and to discriminate the efficacy of barrier creams. PMID- 9734826 TI - Fluorescence spectroscopic investigation of effect of excipients on epidermal barrier and transdermal systems. AB - Excipients are often used in transdermal formulations to overcome the formidable barrier offered by the epidermis in order to achieve the target flux. In this study we describe the use of frequency-domain fluorescence spectroscopy to characterize the effect of two commonly used excipients, propyleneglycol monolaurate (PGML) and oleic acid on stratum corneum and in a silicone-based transdermal delivery system. Fluorescence lifetime and limiting anisotropy for the probe 1,6-diphenyl-1,3,5-hexatriene in isolated human epidermis were measured as a function of formulation treatment. The drop in lifetime ranged from 0.5 to 2.3 ns, indicative of an increased dielectric constant of the lipophilic barrier exposed to all formulations. This increase is due to increased partitioning of the polar excipients from the delivery system into the stratum corneum. The limiting anisotropy showed a drop of 0.1 in the case of the epidermis exposed to oleic acid formulation and not the PGML formulations, indicative of the different modes of action for these two excipients. The fluorescence data suggested fluidization of the silicone matrix by both oleic acid and PGML. The dynamic fluorescence measurements described in this study are a powerful way to screen formulations while gaining valuable mechanistic insight into the mode of flux enhancement in transdermal formulations. PMID- 9734827 TI - The selection and design of topical and transdermal agents: a review. AB - One of the major problems in topical and transdermal drug delivery is the efficiency of the barrier property of the stratum corneum. Topical and transdermal agents were often originally designed as drugs to be given orally. In this paper a mechanistic evaluation of skin penetration shows that topical and transdermal drugs should be designed using different strategies. The relative effects of basic physicochemical parameters are examined. An understanding, at a molecular level, of the permeation process will enable us to produce more effective topical agents and to extend the repertoire of transdermal drugs. PMID- 9734829 TI - Theory of skin electroporation: implications of straight-through aqueous pathway segments that connect adjacent corneocytes. AB - Previous in vitro experiments have shown that transdermal high-voltage pulses (Uskin approximately 100 V; duration approximately 1 ms) create local transport regions (LTR) away from appendages in human skin. Quantitative interpretation of the associated ionic and molecular transport led to the view that a large number of aqueous pathways were created, and these connect the corneocytes within an LTR. Here we use the "brick wall" model of the stratum corneum, modified so that morphology important to understanding electrical behavior is emphasized. In this model a minimum-size LTR is regarded as an idealized stack of corneocytes in which the 5-6 multilamellar lipid bilayer membranes between adjacent corneocytes are electroporated. As in artificial planar bilayer and cell membrane electroporation, a distribution of pathway sizes is expected during pulsing, and during recovery after pulsing individual pathway segments are expected to shrink and close randomly, with a time constant tau(seg) that depends on temperature and on lipid composition. Numerical simulations based on stochastic closure of individual segments were used to predict the electrical conductance G(LTR)(t) of a minimum-size LTR after pulsing stops. These theoretical results show that simple exponential decay, G(LTR)(t) = G(LTR)(0)exp(-t/tau(seg)), occurs with minimal fluctuations if the number of pathways is large (np > 10(2)), but for much smaller values the conduction decreases erratically. A "stochastic bottleneck" leading to complete closure is reached only at about np < 3. Thus, for the same number of electrically created pathways, the stratum corneum will remain "open" longer if the pathways are located within an LTR than if the same number of pathways are distributed sparsely over the skin. These predictions are relevant to postpulse transport, including the trapping of linear macromolecules that can hold pathway segments open for prolonged intervals. PMID- 9734828 TI - Visualization and quantitation of iontophoretic pathways using confocal microscopy. AB - Laser scanning confocal microscopy (LSCM) has been used to visualize and quantitate the penetration of a model, anionic, fluorescent compound (calcein) along the iontophoretic transport pathways within hairless mouse skin. The LSCM technique permits optical sectioning of full-thickness, unfixed tissue, thereby avoiding poor image quality due to blurring from out-of-focus fluorescence, and obviating artifactual redistribution of the permeant during processing. Simple measurements of the approximate flux of the probe across hairless mouse skin were also made using standard in vitro diffusion cell methodology and a fluorometric assay. Most importantly, LSCM imaging strategies were developed to overcome depth dependent sensitivity problems. These visualization studies showed that iontophoresis of calcein into hairless mouse skin enhanced delivery, particularly via follicular structures, to significant depths into the barrier. Nonfollicular transport was also apparent, especially at more superficial levels. Quantitative analyses of the LSCM images showed that, although significant nonfollicular transport occurs, the efficiency of the follicular pathway, when the relative surface area is taken into account, is considerable. Overall, therefore, this work contributes significantly to the ultimate goal of fully comprehending the mechanism(s) of iontophoretic drug delivery across the skin. PMID- 9734830 TI - Skin appendageal macropores as a possible pathway for electrical current. AB - The electrical properties of the outermost layer of skin are described by lipid corneocyte (Zm) and appendageal (Za) impedance, which are connected in parallel. Appendageal macropores are considered as long tubes with distributed electrical parameters. It has been shown that not only Za, but also the macropore resistance Ra and capacitance Ca are frequency dependent. The input of Za in the overall impedance (Z) depends on the space density of active (conductive) macropores n(i), which increase with current density (i) and the duration of iontophoresis. Skin impedance has been demonstrated to decrease under the influence of iontophoretic treatment. Application of the theoretical model to these data provides an estimate of the increase in macropore density during iontophoresis. A comparison of these results with n(i), which was measured directly, shows a strong correlation supporting this unique model. PMID- 9734831 TI - Electroperturbation of human stratum corneum fine structure by high voltage pulses: a freeze-fracture electron microscopy and differential thermal analysis study. AB - Application of high voltage pulses (HVP) to the skin has been shown to promote the transdermal drug delivery by a mechanism involving skin electroporation. The aim of this study was to detect potential changes in lipid phase and ultrastructure induced in human stratum corneum by various HVP protocols, using differential thermal analysis and freeze-fracture electron microscopy. Due to the time involved between the moment the electric field is switched off and the analysis, only "secondary" phenomena rather than primary events could be observed. A decrease in enthalpies for the phase transitions observed at 70 degrees C and 85 degrees C was detected by differential thermal analysis after HVP treatment. No changes in transition temperature could be seen. The freeze fracture electron microscopy study revealed a dramatic perturbation of the lamellar ordering of the intercellular lipid after application of HVP. Most of the planes displayed rough surfaces. The lipid lamellae exhibited rounded off steps or a vanished stepwise order. There was no evidence for perturbation of the corneocytes content. In conclusion, the freeze-fracture electron microscopy and differential thermal analysis studies suggest that HVP application induces a general perturbation of the stratum corneum lipid ultrastructure. PMID- 9734832 TI - Skin electroporation causes molecular transport across the stratum corneum through localized transport regions. AB - High voltage pulsing of human skin (approximately 100 V across the skin, 1 ms pulses) has been hypothesized to cause electroporation of the stratum corneum, and to cause large fluxes of drugs and other molecules across the skin, through newly created aqueous pathways. In contrast, iontophoresis (<0.5 mA per cm2, <1 V across the skin) has long been used in transdermal drug delivery, and is believed to involve pre-existing pathways associated with hair follicles and sweat ducts. Either high voltage pulsing or iontophoresis was applied to human, hairless rat, or black rat snake skin. Hairless rat skin contains more hair follicles than human skin, and snake skin does not contain any hair follicles. All three types of skin had comparable electrical resistances at low voltages; however, the iontophoretic transport of charged fluorescent molecules was significant for human and hairless rat skin, but no transport occured across snake skin, indicating that hair follicles and sweat ducts play a major role in iontophoresis. Electroporation caused large molecular transport for all three types of skin, and involved spontaneously forming localized transport regions, not associated with appendages. These experiments thus provide further support for the hypothesis that high voltage pulsing causes electroporation in the stratum corneum, and that this transport mechanism is fundamentally different from iontophoresis. PMID- 9734833 TI - Light microscopy of living tissue: the state and future of the art. AB - Since its introduction over a decade ago, confocal microscopy has found wide applicability in the microscopy of thick specimens and living tissue because of its ability to obtain images from deep inside the sample without interference from scattered or out-of-focus light. Three new instruments that are capable of imaging optically dense specimens such as the skin are considered here: a modified tandem-scanning confocal, the video-rate laser-scanning confocal microscope (both of which were developed specifically to examine skin in vivo), and the two-photon laser-scanning microscope, a design which is "inherently confocal." The tandem-scanning and video-rate confocals use visible and infra-red light, respectively, to acquire reflection images, whereas the two-photon scanner is a fluorescence microscope. The advantages and drawbacks of each of these instruments is considered. PMID- 9734834 TI - Age-related functional and structural changes in human dermo-epidermal junction components. AB - Cultured normal human keratinocytes obtained from 14 facial skin biopsies of donors aged 9-79 y were used to study the influence of donor age on the integrin receptors, cell adhesive properties in vitro, and type VII collagen synthesis. Immuno-spectrofluorimetric quantitation of integrins showed a decrease in the beta1- and beta4-subunits in low (0.08 mM) and high (1.8 mM) calcium conditions with aging. Calcium ions decreased the fluorescence intensity by relocating integrins at cell boundaries. Measurements of adhering cells showed that adhesion to bovine serum albumin-, type IV collagen- or laminin 1-coated plastic surfaces initially increased until donor age reached 30 y and then decreased. Specific adhesion to type IV collagen and laminin 1 did not vary with age, but the increase in adhesion to type IV collagen produced by manganese ions increased with age, suggesting an age-dependent feature of beta1 integrin. Synthesis of type VII collagen, increased or not by TGFbeta1 (10 ng per ml), did not vary with the donor age. Global normalized principal component analysis showed that variables related to integrins were strongly correlated, as were those of adhesion. Pre-embedding immunoelectron microscopy of freshly isolated keratinocytes showed that certain hemidesmosomes from aged cells had little or no reaction with anti-beta4-chain antibody. Post-embedding type IV collagen immunostaining and image analysis showed less type IV collagen in adult dermo epidermal junctions. These findings indicate that there are structural and functional changes in the dermo-epidermal junction components with aging, probably giving a less effective epidermal anchoring system. PMID- 9734836 TI - Second free flaps in head and neck reconstruction. AB - Over the past decade, free-tissue transfer has greatly improved the quality of oncology-related head and neck reconstruction. As this technique has developed, second free flaps have been performed for aesthetic improvement of the reconstructed site. This study evaluated the indications for and the success of second free flaps. Medical files for patients who underwent second free flaps for head and neck reconstruction at the University of Texas M.D. Anderson Cancer Center, from May 1, 1988 to November 30, 1996, were reviewed. The flaps were classified as being either immediate (done within 72 hr) or delayed (done within 2 years) reconstructions. Indications, risk factors, recipient vessels, outcome, and complications were analyzed. Of the 28 patients included in this study, 12 had immediate (nine as salvage after primary free flap failure, and three for reconstruction of a soft-tissue defect), and 16 had delayed second free flaps (two for reconstruction of a defect resulting from excision of recurrent tumors, and 14 for aesthetic improvement). Reconstruction sites included the oral cavity in 18 patients; the midface in six; the skull base in two; and the scalp in two. The success rate for the second free flaps was 96 percent. Five patients had significant wound complications. In a substantial number of cases, identical recipient vessels were used for both the first and second free flaps. The authors conclude that second free flaps can play an important role in salvaging or improving head and neck reconstruction in selected patients. In many cases, the same recipient vessels can be used for both the first and second flaps. PMID- 9734835 TI - Mechanistic and quantitative prediction of aminopeptidase activity in stripped human skin based on the HaCaT cell sheet model. AB - HaCaT cell culture sheets were recently demonstrated to be a useful tool to study epidermal metabolism. Here we report on a mechanistic and quantitative correlation between the kinetics of aminopeptidase-based cleavage of L-Ala-4 methoxy-2-naphthylamide (Ala-MNA) in HaCaT sheets versus stripped human skin. Fresh human skin (breast or abdominal) was obtained from cosmetic surgery, tape stripped, and dermatomed. HaCaT sheets were cultured on porous membranes. Diffusion and concurrent metabolism were studied under reflection and permeation conditions. Numerical simulations of simultaneous diffusion and saturable Michaelis-Menten metabolism were based on a physical model and a fixed set of independently obtained parameters (diffusion coefficient D, distance x, partition coefficient P, Michaelis constant Km, maximum metabolic rate Vmax). Under reflection conditions, cleavage of Ala-MNA in HaCaT sheets was very close to stripped skin. In contrast, in permeation studies substrate only permeated through HaCaT whereas passage through stripped skin led to full cleavage of Ala MNA to MNA. All experimental data were in reasonable to excellent agreement with numerically generated data. Differences between HaCaT and stripped skin could be quantitatively and mechanistically explained by the thickness of the metabolically active layer, i.e., approximately 10 microm in HaCaT and approximately 40 microm in stripped skin. Full cleavage of permeating Ala-MNA in stripped skin was predicted to occur within the upper approximately 20 microm of viable epidermis. Thus epidermal aminopeptidase activity may act as an efficient metabolic barrier to fully block the permeation of aminopeptidase labile xenobiotics. Within the settings of this study the kinetics of metabolism in the viable epidermis of skin is predictable from HaCaT sheets. PMID- 9734837 TI - Free vascularized fibular grafts for large bone defects in the extremities after tumor excision. AB - Free vascularized fibular grafts were used in nine selected patients with bone tumors in the involved extremity. There were five locally aggressive tumors and four malignant tumors. Each skeletal defect was longer than 10 cm, and the mean length was 13 cm. Ten grafts of 17.4 cm in mean length were harvested in these nine patients. In two cases with upper-extremity involvement, arthroplasty using the fibular head was the procedure of choice, while interacalary grafting was performed in the lower extremity. Dual grafting was performed for complete defects of the femur. The viability of each graft was demonstrated through survival of the combined skin flaps or with a positive bone scan. Primary bony union was obtained in all cases, and the mean time to union was 5.2 months. In the lower extremity, significant hypertrophy of the graft was seen. Satisfactory functional results were obtained in all the patients. PMID- 9734838 TI - Complications in wrap-around-flap donor sites after reconstruction using an arterialized venous flap. AB - The authors have previously reported on the use of the arterialized venous flap to cover skin defects of the big toe resulting from wrap-around-flap transfers. Nineteen patients who had undergone this procedure were reviewed, to determine the incidence of donor-site complications. Problems identified included delayed wound healing, skin erosion, painful callosity, paresthesia, cold intolerance, and cosmetic appearance However, the functional deficit was not great, and most patients were not troubled by their symptoms. The cosmetic appearance of the donor foot was acceptable in most of the patients. PMID- 9734839 TI - An axial-pattern skin flap in the rat. AB - An axial-pattern rat skin flap based on the caudal femoral artery is described with orthotopic and heterotopic island flap transfers. Free-tissue transfers could also be carried out based on the common femoral artery. Since the pedicle is very small, instead of selective dye injection, a nonselective dye injection was utilized, elevating a very large island skin flap based on the caudal femoral artery. This flap model can provide an alternative opportunity to study the physiology of skin flaps and for training in the basic principles of plastic surgery, including microsurgical practice, with a more challenging model. PMID- 9734840 TI - Effect of time on the viability of ischemic skin flaps treated with vascular endothelial growth factor (VEGF) cDNA. AB - This study examined the efficacy of gene therapy on wound healing. The authors investigated whether delivery of the gene encoding a particular cytokine, known to be important in angiogenesis, could affect ischemic skin flaps. Anterior abdominal skin flaps, based solely on the epigastric artery and vein, were created in the Sprague-Dawley rat model. At the time of elevation, the arterial pedicle supplying each flap was infused either with the gene for vascular endothelial growth factor (VEGF) or physiologic saline alone. The flaps were resutured into place and observed for a period of either 4 or 3 days, at which time the pedicle was ligated. Twenty minutes following ligation, blood flow in the flaps was measured by dye fluorescence. Tissue viability of the flaps was subsequently measured by planimetry after a period of 7 days. Flaps that received the VEGF gene and were ligated at 4 days had an average dye fluorescence index (DFI) of 31.1 following ligation, and 93.9 percent viable tissue after 7 days. Flaps that received saline alone, and were ligated following a similar interval, had an average DFI of 14.0 and 31.9 percent viable tissue. Among the subjects that were ligated at 3 days, only a single, gene-infused flap had any noticeable viable tissue after 7 days. The DFI of these groups was 11.0 for the gene-infused group and 22.1 for the saline-infused group. The results suggest that delivery of the gene for VEGF can improve the survival of ischemic skin flaps, but that the effect of gene therapy is not limitless. PMID- 9734841 TI - Intraosseous repair of the inferior alveolar nerve in rats: an experimental model. AB - A reliable method of exposure of the inferior alveolar nerve in Wistar rats has been developed, to allow intraosseous repair with two microsurgical techniques under halothane inhalational anaesthesia. The microsuturing technique involves anastomosis with 10-0 nylon sutures; a laser-weld technique uses an albumin-based solder containing indocyanine green, plus an infrared (810 nm wavelength) diode laser Seven animals had left inferior alveolar nerve repairs performed with the microsuture and laser-weld techniques. Controls were provided by unoperated nerves in the repaired cases. Histochemical analysis was performed utilizing neuron counts and horseradish peroxidase tracer (HRP) uptake in the mandibular division of the trigeminal ganglion, following sacrifice and staining of frozen sections with cresyl violet and diaminobenzidene. The results of this analysis showed similar mean neuron counts and mean HRP uptake by neurons for the unoperated controls and both microsuture and laser-weld groups. This new technique of intraosseous exposure of the inferior alveolar nerve in rats is described. It allows reliable and reproducible microsurgical repairs using both microsuture and laser-weld techniques. PMID- 9734842 TI - Free-flap evolution after hyperthermic regional chemotherapy in the isolated limb for malignant melanoma. AB - Two patients presenting with a stage I melanoma of the sole of the foot (Clark's level IV, Breslow's 2.8 mm, and Clark's level IV, Breslow's 3.2 mm) underwent a 3 cm tumor free-margin skin resection, followed by microanastomosed muscle flap reconstruction (serratus anterior and latissimus dorsi). Immediately after primary wound healing, an elective inguino-iliac lymph-node dissection, followed by hyperthermic isolated regional chemotherapy with Melphalan, was carried out. Only moderate swelling of both free flaps was observed after these procedures, and this resolved rapidly. The patients returned to ambulation after 2 weeks. No other side effects of the hyperthermic isolated regional chemotherapy were observed in the previously microanastomosed flaps. PMID- 9734843 TI - Little finger-to-thumb microvascular transfer. AB - This case report documents a unique thumb reconstruction performed at the authors' institutions. The amputated dominant right thumb of a manual laborer was electively reconstructed with microvascular transfer of the previously partially amputated little finger from the same hand. At 1 year postoperatively, the patient returned to work with excellent grip and pinch strength, thumb opposition to all digits, and 8 mm of static two-point discrimination. The technical details of the operation are described and compared with other analogous reports in the literature. PMID- 9734844 TI - Free-flap distal arteriovenous fistula: when to close it? AB - A distal arteriovenous fistula was created secondarily in an innervated radial forearm free flap, to salvage the neophallus in three female-to-male transsexuals. This resulted in permanent edema and an arterial thrill in the neophallus. The arteriovenous fistulas were closed after 6 weeks to 20 months. Acute endothelial damage with risk of thrombosis, due to ischemia and exposure to arterial blood pressure, may be expected to have been repaired 4 to 6 weeks after arterialization of the veins. Intimal thickening will have started by then, but a subsequent decrease in vascular luminal diameter may still be expected to be reversible. The authors conclude that ligation of the distal arteriovenous fistula may safely be performed some 6 weeks after the salvage procedure. PMID- 9734845 TI - Microvascular tissue transfer in a pregnant patient. AB - Pregnancy is a relative contraindication for elective surgery. The primary concerns are for the safety of the fetus and the mother. However, there are particular problems involving microvascular surgery due to the pregnancy associated hypercoagulable state. The authors were presented with a 35-year woman, 20 weeks pregnant, with a degloved foot and ankle associated with an open distal tibia/fibula fracture (Gustilo IIIB). Salvage of her leg required a microvascular tissue transfer. Accordingly, a combined latissimus dorsi-serratus anterior free flap was performed with a saphenous vein graft to the popliteal vessels. The patient was hypercoagulable and there were extensive platelet clots. Her consumption of heparin was enormous. Postoperatively, she was treated with intravenous dextran for 5 days and for 17 days with intravenous heparin. After discharge, she was placed on subcutaneous heparin until she delivered a healthy baby. The flap survived and her leg was salvaged. The hypercoagulable state of pregnancy, as well as thromboprophylaxis, are discussed. PMID- 9734846 TI - Relationship between cutaneous pressure threshold and two-point discrimination. AB - The amount of pressure that should be applied when doing the two-point discrimination test has always been a matter of controversy. The Pressure specified Sensory Devices permits recording the pressure at which two-point discrimination (2 PD) occurs. The purpose of this study was to investigate the relationship between the cutaneous pressure threshold and 2PD in people with normal and abnormal peripheral nerve functions. The Pressure-specified Sensory Devices was used to quantify the cutaneous pressure threshold in the index-finger pulp in each individual, between the range of 2 mm and 8 mm of static 2 PD, using 1-mm intervals. Twenty normal controls were examined; ten patients were less than 45 years of age; and ten patients were greater than 45 years of age. This relationship of pressure to 2PD was also tested in eight patients with abnormal peripheral nerve function (four patients with carpal tunnel syndrome, and four patients with diabetic neuropathy). A curvilinear relationship was identified in which, for the same skin surface in the same individual, regardless of age or presence of nerve compression or neuropathy, the cutaneous pressure threshold was inversely related to static 2PD. This curve shifted upward and to the right with the increasing age of the normal population and with neurologic impairment. The awareness of this neurophysiologic relationship between 2PD and pressure threshold permits the design of strategies for sensibility testing and provides a basis for the interpretation of sensory test results. PMID- 9734847 TI - Ultrastructure and cellular biology of nerve regeneration. AB - Hippocrates provided the first written description of the peripheral nervous system (PNS), as early as the 4th century B.C., and later Herophilus identified nerves as such, distinguished them from tendons; he also traced nerves to the spinal cord. The traditional Hippocratic teaching of the time, however, doubted that nerve healing occurred. Through the subsequent centuries, several papers were written about the PNS but, without sufficient understanding of anatomy, physiology, and the regenerative capacity of the PNS, it is not difficult to comprehend the frustration that might have been encountered by surgeons in dealing with nerve injuries and their subsequent repair. This was probably the reason why nerve repair was rarely actually undertaken prior to the 19th century. A plethora of studies on the PNS and its regeneration has been reported over the last 150 years and has provided us with current knowledge. It is important, before describing the most recent developments in the area of peripheral nerve regeneration, to briefly outline the major advances over the last century. Currently, the therapeutic approaches taken toward the patient with peripheral nerve injury change continuously. Sophisticated advances in technology, cellular and molecular neurobiology, and electron microscopy will doubtless optimize reconstructive strategies in treating nerve injury. A greater awareness and understanding of the nerve ultrastructure, as well as the underlying mechanisms of the regenerative process and those factors detrimental to nerve regeneration, will assist in the successful repair of nerve injury. This paper reviews the cellular, biochemical, and ultrastructural elements of nerve injury and repair, and the rationale for current reconstructive strategies and techniques. PMID- 9734848 TI - Effects of extruded corn or grain sorghum on intake, digestibility, weight gain, and carcasses of finishing steers. AB - We conducted two trials to evaluate the effects of extruding vs dry-rolling either corn or grain sorghum on intake, digestibility, and performance of finishing steers. In Trial 1, 92 crossbred steers (average BW 413 kg) were used in a 2 x 2 factorial design. Diets contained either dry-rolled corn (RC), extruded corn (EC), dry-rolled grain sorghum (RGS), or extruded grain sorghum (EGS). Diets were fed for 110 d and contained 78.6% of the respective grain, 9% alfalfa pellets, 8.2% molasses, and 4.2% protein-mineral supplement. Daily gain was highest (P < .049) for steers fed RC, and the ADG of steers fed RGS was higher than that of steers fed extruded diets; there was no difference in ADG between steers fed EC and those fed EGS. Steers fed dry-rolled diets consumed more DM (P = .001) than steers fed extruded diets. Feed efficiency was not affected (P = .18) by processing method, but steers fed corn utilized the diets more efficiently (P = .006) than steers fed grain sorghum. Except for carcass weight, carcass data were not affected by grain type (P > .20). Dressing percentage, quality grade, and longissimus muscle area were lower (P < .09) in steers that received extruded grain than in those that received dry-rolled grain. In Trial 2, five ruminally cannulated crossbred steers (average BW 518 kg) were used in a 4 x 4 + 1 Latin square design to evaluate the ruminal and total tract digestion characteristics of the diets used in Trial 1. Type of grain had no effect (P > .16) on intake, total tract digestibility, or ruminal pH. Extruding corn or grain sorghum decreased intake (P < .001) but increased (P < .074) DM and starch digestibility compared with dry rolling; steers fed extruded diets had lower (P < .032) ADF and NDF digestibilities. Ruminal in situ DM and starch disappearance were higher (P < .03) and ruminal pH was lower (P < .052) in steers fed extruded grains than in those fed dry-rolled grains. Data from this study indicate that extruded corn and extruded grain sorghum are highly degradable feeds; however, decreased DM intake and lower ruminal pH levels resulted in lower performance. PMID- 9734849 TI - Development of adjustment factors for backfat and loin muscle area from serial real-time ultrasonic measurements on purebred lines of swine. AB - We recorded serial real-time ultrasonic measurements of backfat and loin muscle area to assess the rate of change per unit of live weight and growth rate measured as the change in weight per day of age during the finishing stage of production. Barrows (648) and gilts (459) representing eight major U.S. pure breeds of swine were evaluated in the study. Real-time ultrasonic measurements of backfat and loin muscle area were collected at average live weights (LW) of 67.4, 80.3, 93.4, and 104.9 kg. Backfat was measured at the midline over the last rib (LR) and last lumbar vertebra (LL) and off the midline at the 3/4 point (BF10) over the 10th rib. Loin muscle area (LMA) images were collected from the right hand side of each pig at the 10th rib. Deposition rates were calculated on an intrapig basis for the dependent scan variables BF10, LMA, LR, and LL in models that included the independent variables LW and LW2. For growth rate (WDA), LW was regressed on age and age. Intrapig linear regression coefficients and y intercepts were further analyzed as dependent variables in a mixed model that included the fixed effects test group, sex, and breed and random effects of sire and dam nested within breed. Significant breed effects were present for deposition rates of BF10, LMA, LR, LL, and WDA. Across the eight breeds, BF10 was deposited at a mean of .271 +/- .008 mm/kg, a rate faster than the average for LR (.201 mm/kg) and LL (.206 mm/kg). The LMA deposition rate across breeds averaged .304 cm2/kg. Average WDA across the breeds was .774 kg. Barrows deposited fat faster at all locations, deposited muscle slower, and grew faster than gilts in this study (P < .001). The results of this study indicate a need for breed- and sex-specific adjustment factors for rates of backfat and loin muscle deposition and for weight per day of age in swine. PMID- 9734850 TI - Does feed restriction mimic the effects of increased ambient temperature in lactating sows? AB - We evaluated the effects of high ambient temperature and feed restriction in primiparous lactating sows. Females were exposed to either a constant thermoneutral (20 degrees C) or hot environment (30 degrees C). Lactating sows at 30 degrees C were given free access to feed (30AL; n = 12), and sows at 20 degrees C were restricted according to the feed intake recorded at 30 degrees C (20RF; n = 6) or were given free access to feed (20AL; n = 6). Jugular vein catheters were surgically inserted at 100 +/- 1 d postcoitum. During lactation, 30AL sows exhibited higher rectal temperatures (P < .05) than 20AL and 20RF sows. Feed intake was reduced by 43% for 30AL compared with 20AL sows. Daily body weight loss was lower (P < .05) in the 30AL than in the 20RF group, and mean litter daily gain over the whole lactation was 18% lower in 30AL than in 20AL sows (P < .05) and was intermediate in 20RF females. Plasma concentrations of thyroid hormones (triiodothyronine [T3] and thyroxine [T4]) were lower at 30 degrees C than at 20 degrees C at d 4 postpartum and d 8 after weaning for T4 (P < .001) and at d 4 postpartum (P < .001) and at d 1 and d 8 after weaning for T3 (P < .01) but were not influenced by feed restriction at 20 degrees C. Mean concentrations of cortisol measured on d 4 and 19 postpartum and on d 1 after weaning were lower in the 30AL than in the 20AL group (P < .05), and neither was different from that in 20RF sows. Ambient temperature and feed intake had no influence on prolactin concentrations on d 19 postpartum and d 1 after weaning. In the 30AL group, concentrations of T3, cortisol, and prolactin measured at d 19 postpartum were positively correlated with the litter gain observed during the 2nd and 3rd wk of lactation (P < .05). The return to estrus was slightly delayed in 20RF compared with 20AL sows (P < .05) and was quite variable in the 30AL group. These results demonstrate that high ambient temperature has negative consequences on litter growth and return to estrus and induces plasma hormonal variations, that cannot be fully mimicked by feed restriction in primiparous sows. PMID- 9734851 TI - Effects of fish meal and sodium bentonite on daily gain, wool growth, carcass characteristics, and ruminal and blood characteristics of lambs fed concentrate diets. AB - We evaluated the effects of replacing some soybean meal (SBM) protein with fish meal (FM) protein in diets adequate and slightly deficient in CP, with or without .75% sodium bentonite (NaB) on performance and ruminal and blood metabolites of individually fed Suffolk lambs. Diets were based on corn, SBM, and cottonseed hulls. In Exp. 1, five lambs were assigned to each of the three dietary treatments (11% CP with 3% FM, 13% CP with 0 or 3% FM). Lambs fed diets that contained 11% CP with 3% FM or 13% CP with 0% FM had similar DMI and ADG. Gain and feed efficiency were slightly improved (P = .18) by the 13% CP diet with 3% FM. In Exp. 2, 32 lambs were assigned to four dietary treatments (13.5% CP of DM) in a 2 x 2 factorial arrangement (0 or 3% FM, and 0 or .75% NaB on an as-fed basis). The DMI and ADG were increased (P < .05) by FM and NaB supplementation. Interactions (P < .05) revealed that NaB increased DMI, ADG, gain per feed (g/kg of DMI), and plasma urea N concentration in the absence of FM but not in the presence of FM in the diet. Neither FM nor NaB influenced (P = .25) wool growth. Total ruminal VFA were increased (P < .06) by FM and NaB. Differences in mineral content of phalanx bone, liver, and kidney were small and may be related to the mineral content of diets and the effect of NaB on mineral solubilities. Similar DMI and ADG of lambs fed FM and NaB separately and in combination suggest that their beneficial effect is not additive. PMID- 9734852 TI - Fertility-associated antigen on bull sperm indicates fertility potential. AB - A 30-kDa heparin-binding protein named fertility-associated antigen (FAA) was identified in sperm membranes of beef bulls with greater fertility potential. In a survey of 2,191 beef bulls, 88% had FAA present in sperm membranes (FAA positive), and 12% were FAA-negative. In the first study, 54 Santa Gertrudis and 51 Santa Cruz bulls were grouped (1 to 14 bulls per group) according to FAA profiles and were bred to 2,403 cows at ratios of 1 bull: 25 cows. Fertility for 14 groups of FAA-positive bulls averaged 88%, whereas three groups of FAA negative bulls impregnated 79% of the cows. Thus, FAA-positive bulls were nine percentage points more (P < .01) fertile than FAA-negative bulls. In the second study, 2-yr-old Santa Cruz bulls (n = 26) were grouped according to FAA profiles and serving capacity. The fertility of the group of 12 high-serving-capacity, FAA positive bulls was 87% of 270 cows. The group of six FAA-negative bulls with high serving capacity impregnated 78% of 143 cows. Among the groups of bulls with high serving capacity, FAA-positive bulls were nine percentage points more (P < .05) fertile than FAA-negative bulls. The group of eight FAA-positive bulls with low serving capacity impregnated the least (P < .01) percentage (69%) of 238 cows. Serving capacity of bulls should be considered when optimizing fertility potential. Among bulls with acceptable physical characteristics and serving capacity, determination of FAA profiles in sperm can be used as a tool to identify subfertile bulls. PMID- 9734854 TI - Bayesian analysis of twinning and ovulation rates using a multiple-trait threshold model and Gibbs sampling. AB - The Multiple-Trait Gibbs Sampler for Animal Models programs were extended to allow analysis of ordered categorical data using a Bayesian threshold model. The algorithm is based on data augmentation, where a value on the unobserved underlying normally distributed variable (liability) is generated in each round of iteration for each categorical observation. The programs allow analysis of several continuous and ordered categorical traits. Categorical traits can have any number of response levels. Models can be different for each trait. The programs were used to analyze twinning and ovulation rates from a herd of cattle selected for twinning rate at the U.S. Meat Animal Research Center. Data included number of calves born at each parturition for the lifetime of a cow and number of eggs ovulated for several estrous cycles before first breeding as heifers. A total of 6,411 calvings was recorded for 2,087 cows with 83.2% single and 16.8% multiple births. A total of 19,849 ovulations was recorded for 2,332 heifers with 85.2% single and 14.8% multiple ovulations. Mean posterior estimates of heritability and fraction of variance accounted for by permanent environmental effects (PE) were .128 and .103 for twinning rate and .168 and .079 for ovulation rate. Mean posterior estimate of genetic correlation was .808, and correlation of PE effects was .517. Use of a threshold model could allow for more rapid genetic improvement of the twinning herd through improved identification and selection of genetically superior animals because of higher heritability on the underlying scale. PMID- 9734853 TI - Factors contributing to the incidence of dark cutting beef. AB - The 1995 National Beef Quality Audit reported that dark cutting beef (dark cutters) cost $6.08 per animal harvested in the United States. Feedlot data were obtained over a 3-yr period from nine commercial feedyards (15,439 pens of cattle; 2,672,223 total cattle). Feedyard, sex, implant treatment, days from final implant to harvest, maximum and minimum daily temperatures, and temperature fluctuations from 2 d before harvest to the day of harvest all contributed (P < .05) to the incidence of dark cutters. Heifers yielded a higher (P < .05) percentage of dark cutters per pen and, when reimplanted a second time with an estrogenic implant, produced greater (P < .05) mean percentages of dark cutters per pen than heifers reimplanted with either androgens or combination (androgen and estrogen) growth promotants. Furthermore, heifers produced higher (P < .05) mean percentages of dark cutters per pen than steers during periods of hot (> 35 degrees C) weather 2 to 1 d before harvest. Steers, when treated with a combination (androgen and estrogen) implant when entering the feedyard and as a reimplant, produced higher (P < .05) mean percentages of dark cutters per pen when compared to other moderate growth-promoting implant strategies. When producers opted to implant steers with estrogenic growth promotants, either as the cattle entered the feedlot or as a final reimplant before harvest, the occurrence of dark cutters was reduced from 9.2 per thousand cattle shipped to 2.0 and .5 per thousand cattle shipped, respectively. Producers that reimplanted heifers before harvest with products that were not primarily estrogenic reduced the occurrence of dark cutters from 10.4/1,000 cattle shipped to 5.2/1,000 cattle shipped when androgen-based growth promotants were used and to 3.5/1,000 cattle shipped when combination (androgen and estrogen) implants were administered. In addition to implant selection, those producers that held cattle on-feed over 100 d past reimplantation reduced the incidence of dark cutters per pen by an average of 38% among heifers and 69% among steers. By reducing the occurrence of dark cutters, there is an opportunity for beef producers to realize large economic savings. PMID- 9734855 TI - Evaluation of the ovine callipyge locus: I. Relative chromosomal position and gene action. AB - Genotypic and phenotypic data were collected to estimate chromosomal position of the callipyge (CLPG) gene and to test gene action. Nine Dorset rams of extreme muscling phenotype and 114 Romanov ewes composed the grandparent generation of a resource flock of 362 F2 lambs segregating at the CLPG locus. The parent generation consisted of eight F1 sires and 138 F1 dams. The F2 lambs were serially slaughtered in six groups at 3-wk intervals starting at 23 wk of age to allow comparisons at different end points. A linkage group of 25 marker loci (mean of 708 informative meioses per marker) spanning 87.2 cM was developed and improved the previous known coverage and precision of marker order and interval distance from available maps of ovine chromosome 18. Probabilities of each CLPG genotype were calculated at 1-cM intervals (0 to 107 cM). Statistical models included effects of year, sex, sire, regressions on genotypic probabilities, and genotype-specific linear and quadratic regressions on appropriate covariates. Orthogonal contrasts of CLPG genotypic effects evaluated additive, maternal dominance, and paternally derived polar overdominance models of gene action. The most parsimonious model did not include the additive and maternal dominance genetic contrasts. From analyses of four key traits, a consensus for position of CLPG was obtained at 86 cM relative to the most centromeric marker. An F-test with 3 df representing polar overdominance was maximum at position 86 cM (F = 407.4; P < .00001) with leg score as the dependent variable. These results are consistent with assignment of the CLPG locus to the telomeric region of chromosome 18 and support the polar overdominance model of gene action proposed by Cockett et al. (1996). Furthermore, recombinant individuals with definitive phenotypes confined the position of CLPG to a 3.9-cM interval, facilitating positional cloning experiments. PMID- 9734856 TI - Do domestic animals have minds and the ability to think? A provisional sample of opinions on the question. AB - Faculty, staff, and graduate students in a number of departments, students in an undergraduate course, and some groups outside the university were polled to obtain their perceptions about whether domestic animals have minds, the ability to think, and differing degrees of intelligence (the surveys focused only on horses, cows, sheep, dogs, chickens, pigs, cats, and turkeys). A clear majority of all groups surveyed (except the Department of Zoology) said yes, they believe animals have minds, but a substantial number of those in animal sciences and zoology (17 to 25%) said no. A number of others in animal sciences, zoology, and philosophy (11 to 37%) refused to answer the question because the concept of mind was not defined. From 80 to 100% of respondents in other groups said yes to the question of minds. From 67 to 100% of all participants said yes, they perceive that animals have the ability to think, but a substantial number of animal scientists, zoologists, veterinarians, and English faculty said no, animals don't think (6 to 33%). On the question Do domestic animals differ in relative intelligence?, the responses varied from 88% in animal sciences to 100%. Surprisingly, when asked to rank different animal species by intelligence, there was a remarkable degree of similarity across all groups regardless of background; the overall ranking from highest intelligence to lowest was dog, cat, pig, horse, cow, sheep, chicken, and turkey. Most of the respondents believed that the possession of minds, thought, and intelligence were relevant factors in how animals should be treated and the prevalent concept was that we should not be cruel to animals, but should treat them humanely. PMID- 9734857 TI - Effects of regular moving and handling on the behavioral and physiological responses of pigs to preslaughter treatment and consequences for subsequent meat quality. AB - The effects of regular moving and handling during the finishing period on behavioral and physiological responses of pigs during preslaughter treatment and consequences for meat quality were studied. From the age of 10 wk onward, 144 pigs were housed in groups of four (two gilts and two castrates) and subjected to one of the following treatments. The Environment treatment allowed pigs to move freely for 8 min outside their home pen. Then the pigs were transported in a box for 2 min, and after which they were returned to their home pen. In the Handling treatment, an experimenter remained for 3 min in the pen, and whenever a pig made contact, it was gently stroked. The experimenter then walked for an additional 1 min, without attempting to pat or stroke any pigs but subsequently held each pig in a tight grip for about 5 s. This entire procedure was then repeated. A Control treatment was also included, in which the pigs were subjected to no treatment. The Environment and Handling treatments were applied twice a week at the age of 15, 17, 19, 21, and 23 wk. At 25 wk of age, pigs were transported to the abattoir. They were held unmixed in the truck and in lairage and were manually stunned. The stockmen needed significantly less time to move Environment pigs out of their pen and into the transport box. There were no differences between treatments in salivary cortisol concentrations before or after transport. Environment and Handling pigs had paler meat than Control pigs. Glycogen content at 1 h after death and water-holding capacity were lower in Environment pigs than in Control pigs, but this did not lead to a higher incidence of PSE meat. We conclude that the pigs that had experience with leaving their home pen and some of the transport conditions were much easier to handle at loading. Pigs that are easier to move are less likely to be subjected to rough handling, which implies improved welfare, and the workload for personnel at the time of marketing is reduced. Differences in meat quality due to treatment were slight. PMID- 9734858 TI - Increases in insulin-like growth factor binding protein-2 accompany decreases in proliferation and differentiation when porcine muscle satellite cells undergo multiple passages. AB - Subjecting cloned porcine myogenic satellite cells to multiple passages leads to decreased rates of cell division and myotube formation. Because IGF have been implicated in the regulation of muscle cell proliferation and differentiation, the present study was conducted to characterize secretion of IGF-I and IGF binding proteins (IGFBP) in cultures of cloned porcine satellite cells at two stages of multiple passaging. To this end, we obtained a single porcine satellite cell clone that demonstrated relatively high capacities for cellular proliferation and differentiation into myotubes at the fifth passage but that had greatly diminished capacities for proliferation and myotube formation by the seventh passage. The predominant IGFBP secreted by this satellite cell clone was immunologically identified as IGFBP-2, and quantities of it were increased in medium from seventh-passage cultures. Quantities of IGF-I in medium were determined with a newly developed "titration" radioimmunoassay in which interference from IGFBP was minimized by adding a range of saturating quantities of IGF-II. Medium IGF-I concentrations in seventh-passage cultures were also increased relative to the fifth-passage cultures when expressed per unit of DNA. It is hypothesized that the observed increase of IGF-I in medium likely resulted from protective sequestration of IGF-I by IGFBP-2 rather than from enhanced IGF-I secretion. In summary, these data suggest that multiple passaging of cloned porcine satellite cells results in increased secretion of IGFBP-2, which is associated with depressed cell proliferation and myotube formation, perhaps because the increased IGFBP-2 sequestered IGF-I and reduced its bioactivity. PMID- 9734859 TI - An evaluation of current and alternative systems for quality grading carcasses of mature slaughter cows. AB - Strip loins from 354 female bovine carcasses, selected to represent 30 skeletal maturity (A, B, C, D, and E) x marbling score (SA/MA/AB, MD, MT, SM, SL, and TR/PD) subclasses, were used to evaluate current and alternative systems for classifying cow carcasses into expected-palatability groups. Strip loins were vacuum-packaged, stored for 14 d postmortem at 2 degrees C, and frozen (-27 degrees C). Five steaks from each strip loin, each cooked to a different internal temperature (60, 66, 71, 77, or 82 degrees C), were used for shear force determinations. Two steaks from each strip loin, one cooked to 66 degrees C and the other to 77 degrees C, were used for sensory evaluation. Increased carcass maturity was associated with decreased tenderness and juiciness, increased flavor intensity, and a higher incidence of flavors described as "painty," "fishy," and "grassy." Position of a carcass within a maturity group had a negligible effect on palatability. Increased marbling was associated with greater tenderness and juiciness, a lower incidence of steaks with a "grassy" flavor, and a higher incidence of steaks with a flavor described as "fatty." Relationships between marbling and beef palatability traits were consistent across all maturity groups. Carcasses of maturities A through E were most effectively stratified according to differences in palatability when marbling scores were grouped as follows: 1) MD and higher; 2) SL, SM, MT; and 3) TR/PD. Among mature (C, D, and E maturity) carcasses, yellow-colored fat was associated with greater beef toughness and higher detection rates for "grassy" and "fishy" flavors. Higher end-point temperatures were associated with higher shear force values and lower ratings for muscle fiber tenderness, connective tissue amount, overall tenderness, and juiciness. Two alternative grading approaches (one involving current quality grading factors and the other involving the use of fat color as an additional grade factor) were developed for possible use in classification of cow carcasses into expected-palatability groups. Both alternative systems provided a more effective stratification of cow carcasses according to palatability differences than did the current USDA quality grading system. PMID- 9734860 TI - Composition analysis of pork carcasses by dual-energy x-ray absorptiometry. AB - Dual-energy x-ray absorptiometry (DXA) was used as a noninvasive method to measure the composition of pig carcasses. A total of 181 half-carcasses (10 to 51 kg, from pigs slaughtered at approximately 30, 60, 90, and 120 kg) were scanned using a Lunar (Madison, WI) DPX-L densitometer. The DXA measurements of fat, lean, bone mineral, and total tissue mass were compared with chemical analysis for fat, water, protein, total ash, and scale weight. The mean value for total tissue mass by DXA was slightly less than the mean carcass weight (32.3 kg vs 33.6 kg, P > .05, R2 = .998). Although highly correlated (R2 = .81), the DXA measurement of the percentage of fat in the half-carcass was less (P < .001) than the chemical measurement (19.5 vs 24.9%). The DXA measurement of lean tissue mass (total mass less fat and bone mineral) was correlated with carcass protein (R2 = .97) and water (R2 = .99) content. The correlation (R2) between DXA bone mineral content and carcass ash content was only .68; however, DXA bone mineral content was more highly correlated with carcass weight (R2 = .93) than was carcass ash content (R2 = .70). When we used the DXA R value (ratio of the attenuation coefficients for fat and lean) to predict percentage of fat in the carcass, the mean value for predicted carcass fat was 25.9% (P > .05). Similarly, carcass protein and water content were predicted from DXA lean. Using DXA region of interest analysis, estimates of the fat content of the shoulder and ham regions were close to chemical values; however, DXA underestimated the fat content of the loin and side regions by 20 and 28%, respectively. When prediction equations were used to evaluate DXA measurements of the half-carcasses of 28 gilts and 37 boars slaughtered at approximately 120 kg, the half-carcasses of gilts contained more fat (33.9 vs 27.8%, P < .001), less protein (14.1 vs 16.1%, P < .001), and less water (45.9 vs 52.1%, P < .001) than those of boars. These results indicate that DXA could be a valuable research tool for measuring the composition of pig carcasses. On the basis of the results of this study, prediction equations were revised for the DXA estimation of fat, protein, and water content of the half carcass: Fat (%) = 450 - (315 x DXA R value), Protein (g) = -145 + (.23 x DXA lean), and Water (g) = 150 + (.73 x DXA lean). Furthermore, it seems that separate prediction equations are needed for regional analysis. PMID- 9734861 TI - Near-infrared reflectance analysis for predicting beef longissimus tenderness. AB - Near-infrared reflectance spectra (1,100 to 2,498 nm) were collected on beef longissimus thoracis steaks for the purpose of establishing the feasibility of predicting meat tenderness by spectroscopy. Partial least squares (PLS) analysis (up to 20 factors) and multiple linear regression (MLR) were used to predict cooked longissimus Warner-Bratzler shear (WBS) force values from spectra of steaks from 119 beef carcasses. Modeling used the combination of log(1/R) and its second derivative. Overall, absorption was higher for extremely tough steaks than for tender steaks. This was particularly true at wavelengths between 1,100 and 1,350 nm. For PLS regression, optimal model conditions (R2 = .67; SEC = 1.2 kg) occurred with six PLS factors. When the PLS model was tested against the validation subset, similar performance was obtained (R2 = .63; SEP = 1.3 kg) and bias was small (<.3 kg). Among the 39 samples in the validation data set, 48.7, 87.7, and 97.4% of the samples were predicted within 1.0, 2.0, and 3.0 kg, respectively, of the observed Warner-Bratzler shear force value. The optimal PLS model was able to predict whether a steak would have a Warner-Bratzler shear force value < 6 kg with 75% accuracy. The R2 of MLR model was .67, and 89% of samples were correctly classified (< 6 vs > 6 kg) for Warner-Bratzler shear force. These data indicate that NIR is capable of predicting Warner-Bratzler shear force values of longissimus steaks. Refinement of this technique may allow nondestructive measurement of beef longissimus at the processing plant level. PMID- 9734862 TI - Derivation of an equation to estimate marrow content of bovine cervical vertebrae. AB - Marrow content of bovine cervical vertebrae from Choice- and Select-grade carcasses weighing 294 to 343 kg was determined so that a method to monitor the amount of marrow in meat from advanced meat/bone separation machinery and recovery (AMR) systems could be developed. The marrow determination requires cleaning and then ashing bones. Because a large difference in ash content of bone and bone marrow exists and because cartilage content of cervical vertebrae in Choice and Select beef is relatively constant, it was possible to derive the following equation: Weight of marrow = [weight of cartilage (% ash in cartilage - % ash in bone) + % ash in bone (total weight) - (total ash)]/[(% ash in bone - % ash in marrow)]. Constants for ash in fresh bone, marrow, and cartilage were 58.51, .57, and 2.14% with SD of 2.23, .15, and .30%, respectively. A cartilage content of 9.5% along with cervical vertebrae weight and total ash weight were also used to calculate 33.9% marrow in cervical vertebrae. Means for marrow pressed or centrifuged from bovine cervical vertebrae were lower than those obtained from the equation. Therefore, pressing and centrifuging left some marrow in spongy bone. Our ashing method for determining the amount of marrow in whole cervical vertebrae should be useful for determining marrow remaining in cervical vertebrae of bone cakes from AMR systems. Percentage ash in pressed bones is higher and the calculated marrow content is lower when pressed bones are compared to cervical vertebrae that are not pressed. The amount of marrow in whole cervical vertebrae minus the amount left in cervical vertebrae from bone cakes equals the amount in meat from AMR systems. PMID- 9734863 TI - Influence of dietary total sulfur amino acids and methionine on growth performance and carcass characteristics of finishing gilts. AB - We conducted three experiments to determine the sulfur amino acid (SAA) and methionine requirements of finishing gilts. Gilts (PIC Line 326 x C-15, Exp. 1; Line 326 x C-22, Exp. 2 and 3) were blocked by initial weight in randomized complete block designs. In Exp. 1, 64 gilts (initially 54 kg) were fed diets containing either .56 or .44% apparent digestible lysine with increasing SAA levels (63, 70, and 77% of apparent digestible lysine) in a 2 x 3 factorial. A lysine x SAA interaction (P < .10) was observed for ADG and ADFI. Increasing SAA:lysine ratios from 63 to 70% in diets containing .56% apparent digestible lysine increased ADG and ADFI; however, increasing the SAA:lysine ratio in diets containing .44% apparent digestible lysine decreased ADG and ADFI. Pigs fed .56% apparent digestible lysine had higher (P < .05) ADG and gain:feed ratio (G/F) and lower 10th rib fat depth than pigs fed .44% apparent digestible lysine. Increasing the SAA:lysine ratio had no effect on G/F or carcass characteristics. In Exp. 2, 80 gilts (initially 74 kg) were fed diets containing .225, .25, .275, .30, or .325% apparent digestible SAA (45, 50, 55, 60, or 65% of .50% apparent digestible lysine, respectively). Increasing SAA concentrations decreased ADG and G/F (linear, P < .06). In Exp. 3, 105 gilts (initially 72 kg) were fed diets containing .20% apparent digestible cystine and .10, .125, or .15% apparent digestible methionine (20, 25, or 30% of .50% apparent digestible lysine). Increasing digestible methionine increased ADG, ADFI, plasma methionine concentrations (linear, P < .01), and G/F (quadratic, P < .03). The greatest increases in ADG and G/F were observed when apparent digestible methionine was increased from .10 to .125%. Based on these results, the apparent digestible methionine requirement is no greater than 25% of apparent digestible lysine, in diets containing excess cystine. This equates to an apparent digestible SAA:lysine ratio that is no greater than 50%. PMID- 9734864 TI - Effects of a direct-fed yeast culture on enteric microbial populations, fermentation acids, and performance of weanling pigs. AB - In three replicate trials, a total of 36 pigs that had been cannulated at the terminal ileum were used to determine the effects of a Saccharomyces cerevisiae culture in a phase feeding program (phase I was d 0 to 7 and phase II was d 8 to 21) on performance, ileal microflora, and short-chain fatty acids in weanling pigs. Pigs were cannulated at approximately 12 d of age, weaned at 17 d of age, and randomly assigned to one of three treatments: 1) a pelleted phase feeding program, 2) a similar program with the inclusion of a live S. cerevisiae culture (1 g/ kg), and 3) a nonpelleted feeding program otherwise similar to program 2. Ileal samples were collected at 17, 20, 24, 27, 31, 34, and 38 d of age, and samples were analyzed for total E. coli, streptococci, lactobacilli, yeast, short chain fatty acids, pH, and dry matter. Performance data were also collected. At 41 d of age, pigs were killed and digesta were collected from various regions of the gastrointestinal tract. Total intake was less for pigs fed the control diet than for pigs fed the yeast diets, and overall gains tended to be greater for pigs fed diets including yeast. Treatment differences were not observed for ileal microflora or short-chain fatty acids in samples obtained from cannulas or from the various sites of the gastrointestinal tract. Inclusion of a live yeast culture in weanling pig diets affected intake and performance but did not alter tested intestinal microflora or net concentrations of fermentation products. PMID- 9734865 TI - Urinary and biliary excretion of ergot alkaloids from steers that grazed endophyte-infected tall fescue. AB - Ergot alkaloids cause fescue toxicosis when livestock graze endophyte-infected (E+) tall fescue. Little is known about the bioavailability of the ergot alkaloid classes (lysergic acid, lysergic acid amides, or ergopeptine alkaloids) in livestock, and this hampers development of pharmacological strategies to ameliorate the toxicosis. One method used to determine bioavailability of ergot alkaloids is to examine urinary and biliary excretion patterns. Thus, our objectives were to compare ergot alkaloid excretion via urinary or biliary systems and to determine the rate of appearance or clearance of these alkaloids in cattle that were grazing E+ or endophyte-free (E-) tall fescue. In autumn 1996, bile and urine samples were collected from eight steers (203 kg), each grazing E+ and E- tall fescue, and total alkaloid excretion was quantified using competitive ELISA. Approximately 96% of the ergot alkaloids were excreted in urine. The same steers were used to examine the rate of appearance in, or clearance from, urine when switched from E+ to E-, or from E- to E+, pastures in comparison with steers that were continuously grazing E+ or E- tall fescue at 0, 2, 5, and 7 d. Steers were returned to their original pastures after 7 d, and urine was collected at 2, 5, and 7 d. Urinary alkaloid concentrations in steers switched from E- to E+ pastures were similar (P = .55) to those in steers that continuously grazed E+ tall fescue after 2 d. Steers switched from E+ to E- pastures had urinary alkaloid concentrations similar (P = .91) to those in steers that continuously grazed E- pastures after 2 d. In 1997, two trials were conducted in which steers (191 kg) were switched or remained on E+ or E- pastures, and urine was collected at 0, 12, 24, 48, and 96 h to estimate rate of alkaloid appearance or clearance. Steers switched from E- to E+ 1) had about 33% as much urinary alkaloids as steers grazing E+ pasture after 12 h, 2) were not different after 24 h (P = .76), 3) had twice those of the E+ steers at 48 h (P < .05), and 4) were not different after 96 h. Steers switched from E+ to E- tall fescue had approximately 33% less (P < .05) urinary alkaloids than those grazing E+ at 12 h, 67% less (P < .05) at 24 and 48 h, and were not different (P = .86) from steers continuously grazing E- pastures after 96 h. Urinary alkaloid excretion patterns were similar to ergot alkaloid solubility patterns from in vitro digestion of E+ tall fescue. We suggest that alkaloids, liberated from the forage by ruminal microorganisms, were rapidly absorbed as lysergic acid amides and biotransformed ergopeptine alkaloids. PMID- 9734866 TI - A quantitative competitive reverse transcription-polymerase chain reaction technique to measure porcine interferon-gamma. AB - In swine, as in other mammals, interferon-gamma (IFN-gamma) plays an important role in the regulation of the immune response. The objective of this work was to develop a sensitive, quantitative competitive reverse transcription-polymerase chain reaction (qcRT-PCR) assay, using an internal cRNA standard, to monitor the expression of porcine IFN-gamma gene. We applied this qcRT-PCR assay to quantify the expression of IFN-gamma transcripts in peripheral blood mononuclear cells (PBMC) incubated for different times with or without concanavalin A (ConA). Results showed that the expression of the IFN-gamma transcripts in PBMC occurred within the first 2 h after mitogenic stimulation and reached a peak after 24 h of incubation with ConA. The qcRT-PCR technique provides an efficient and sensitive tool for studying the expression of porcine IFN-gamma gene under a variety of experimental conditions. This will help to understand the regulation of T cell mediated immune responses by IFN-gamma at the gene level. PMID- 9734867 TI - Serum concentrations of luteinizing hormone, growth hormone, and cortisol in gilts treated with N-methyl-D,L-aspartate during the estrous cycle or after ovariectomy. AB - The objective of this experiment was to determine the effects of n-methyl-d,l aspartate (NMA), an agonist of the neurotransmitter glutamate, on circulating concentrations of LH, GH, and cortisol in gilts treated during the luteal (n = 4) or follicular (n = 4) phase of the estrous cycle, or after ovariectomy (n = 4). Blood was sampled every 15 min for 10 h on each of two consecutive days. On the 1st d, two gilts from each group received i.v. injections of NMA (10 mg/kg BW) at h 4 and 6, and the remaining gilts received .9% saline (vehicle). The following day, gilts that had received NMA on the 1st d received vehicle, and gilts that had received vehicle on d 1 received NMA. All gilts received an i.v. challenge of GnRH (.1 microg/kg BW) at h 8 on each day. The NMA treatment increased (P < .01) LH pulse frequency in luteal-phase gilts by 125%. In contrast, NMA decreased (P < .05) mean concentrations of LH by 48% and suppressed (P < .01) LH pulse frequency by 33% in ovariectomized gilts. No characteristics of LH secretion were affected (P > .05) by NMA in follicular phase gilts. Serum LH concentrations for the 2-h period following GnRH were lower (P < .05) in follicular-phase gilts than in ovariectomized gilts and were 1.15 +/- .09 (mean +/- SE), .81 +/- .05, and .51 +/ .17 ng/mL for ovariectomized, luteal-phase, and follicular-phase gilts, respectively. Treatment with NMA increased circulating concentrations of GH by 334% (P < .01) and cortisol by 77% (P < .03) in all gilts. We suggest that the effects of NMA on LH release in gilts depend on the circulating steroidal milieu. In contrast, NMA evokes secretion of GH and cortisol irrespective of the reproductive status of treated gilts. PMID- 9734868 TI - The impact of either a Meishan or Yorkshire uterus on Meishan or Yorkshire fetal and placental development to days 70, 90, and 110 of gestation. AB - Straightbred Yorkshire (Y) conceptuses are larger than straightbred Meishan (M) conceptuses throughout gestation and at farrowing. In contrast, when Y and M conceptuses were gestated together in Y recipient females, the birth weight of M pigs was similar to that of their Y littermates. Even though placentae of M pigs remained markedly smaller than placentae of Y littermates, they were significantly more vascular. The objective of this study was to compare and contrast the changes in Y and M conceptus growth and placental-endometrial vascularity throughout late gestation in Y or M uteri. Gravid uteri were recovered at slaughter from M and Y females that were gestating either M or Y conceptuses on d 70, 90, or 110 of gestation. Uterine and conceptus measurements were recorded, and a section of the intact endometrial-placental attachment site for each conceptus was fixed, embedded, and later evaluated for placental and endometrial vascular density. Placental surface area and weight were greater (P < .001) when M or Y conceptuses were recovered from Y uteri compared with M uteri on each day of gestation examined. Further, by d 110, the surface area of Y placentae was greater (P < .001) than that of M placentae, regardless of uterine type in which they were gestated. The vascular density of M placentae and adjacent endometrium doubled (P < .05) between d 70 and 110 of gestation (3.0 and 2.5 vs 6.0 and 5.1%, respectively), with no significant increase in placental surface area. In contrast, the surface area of Y placentae doubled in size (P < .001) between d 90 and 110 of gestation, but placental and adjacent endometrial vascular density remained relatively constant, averaging 3.2 and 3.8%, respectively. These data are consistent with the premise that placental size is largely determined by the uterus in which a conceptus is gestated until approximately d 90. After d 90, fetal breed-specific mechanisms maintain optimal fetal growth. Between d 90 and term, M fetal growth depends on progressive increases in placental blood vessel density and requires no increase in placental size. In contrast, Y conceptuses seem to rely exclusively on placental growth to increase placental-endometrial surface area for nutrient exchange. PMID- 9734869 TI - Effect of nutritional management, trace mineral supplementation, and norgestomet implant on attainment of puberty in beef heifers. AB - We conducted a study to evaluate the influences of nutritional management, trace mineral supplementation, and exogenous progesterone on attainment of puberty in beef heifers. Heifers (n = 180) were assigned at weaning to blocks and treatments. Treatments included two dietary regimens (corn silage vs pasture + oatlage), trace mineral supplementation, and puberty induction strategy (with or without progestin implant). Heifers that received pasture + oatlage were managed on grass-legume pastures from October 14 until December 14 and were then placed in pens and fed an oatlage-based diet through May 1994. Heifers fed the corn silage-based diet were housed in pens throughout the study. Norgestomet was implanted in half of the heifers on April 11 for 10 d. Progestin implant increased (P < .05) the number of heifers that had attained puberty by the end of the study, compared with nonimplanted heifers (89% vs 71%). Trace mineral supplementation did not affect percentage of heifers that reached puberty before the implant period. Plasma copper levels were below recommended levels in heifers fed oatlage-based diets without trace minerals. We conclude that heifers can be placed on regrowth in irrigated pastures during the fall and still make acceptable gains for attainment of puberty the following spring and that progestin treatment can aid in inducing heifers to reach puberty. PMID- 9734870 TI - Insulin and glucagon secretion in lactating cows during heat exposure. AB - Heat stress affects endocrine systems in cows. This study investigated changes in insulin and glucagon secretion between thermoneutral (TN; 18 degrees C, relative humidity [RH] 60%) and hot (28 degrees C, RH 60%) environments in lactating cows. Glucose, arginine, and butyrate were administered i.v. to four cows (mean, at 83 d postpartum) in each environment. Blood was collected via a jugular catheter at regular intervals. Heat exposure resulted in a marked increase in respiration rate and rectal temperature. A decrease in milk yield was also observed during heat exposure. Basal insulin concentrations were elevated, and basal glucose concentrations tended to be lower in the hot environment. Peak values of insulin and glucagon following the arginine injection were significantly higher in the hot than in the TN environment. The insulin peak value in response to the butyrate infusion was also higher during the heat exposure. However, insulin and glucagon responses to the glucose load were not affected by heat stress. The increase in plasma glucose concentration following arginine injection was inhibited by the heat exposure. In conclusion, heat stress resulted in a higher insulin secretion in lactating cows. Glucagon secretion in response to the arginine injection was enhanced, but the rise in plasma glucose was inhibited by heat exposure. These changes would be related to a reduction in milk yield during heat stress. PMID- 9734871 TI - The role of pH in regulating ruminal methane and ammonia production. AB - When steers (n = 4) were fed increasing amounts of concentrate (0, 45, or 90% of DM) and decreasing amounts of forage, the VFA concentration increased (P < .001) and ruminal pH, acetate:propionate ratio, and dissociated ammonia declined (P < .001). Acetate:propionate ratio and dissociated ammonia were highly correlated (r2 = .82 and .65, respectively) with ruminal pH. In vivo acetate:propionate ratio was highly correlated (r2 = .78) with the capacity of the bacteria to produce methane from H2 and CO2 in vitro, and in vivo pH-dissociated ammonia was correlated (r2 = .59) with the capacity of the bacteria to produce ammonia from protein hydrolysate. The role of pH in regulating methane and ammonia production was supported by the effect of pH in vitro. When bacteria from cattle fed concentrate or forage were incubated at pH values from 6.5 to 5.7, methane production decreased (P < .001) from 48 to 7 nmol x mg protein(-1) x min(-1) and from 14 to 2 nmol x mg protein(-1) x min(-1), respectively. The reduction in in vitro pH (6.5 to 5.7) also decreased (P < .001) the rates of ammonia production, but only if the bacteria were obtained from cattle fed forage (28 to 15 nmol x mg protein(-1) x min(-1)). Bacteria from cattle fed 90% concentrate had similar (P > .05) rates of ammonia production at pH 6.5 to 5.7 (approximately 12 nmol x mg protein(-1) x min(-1)). These results indicated that ruminal pH affected ruminal methane production, acetate:propionate ratio, deamination, and ammonia concentration. PMID- 9734872 TI - Kinetics of plasma fructose and glucose when lactose and fructose are used as energy supplements for neonatal calves. AB - Shortly after birth, plasma glucose and fructose concentrations of the neonate decline and thus leave blood sugar below the homeostatic mode. Two trials were conducted to determine the plasma glucose and fructose kinetics in control and supplemented calves for 108 h after birth. In the short-term trial, six Holstein calves were given 40 g of either fructose, lactose, or water (control) orally at 1 and 96 h after birth. Treatments were administered with a colostrum substitute (Life Boost) at 1 h and whole milk at 96 h. Rectal temperatures and changes in plasma glucose and fructose concentrations were monitored at close intervals for 12 h after supplementation. In the long-term trial, 15 Holstein calves were given 40 g of either lactose, fructose, or water (control) at 1 h after birth and at 12 h intervals for 81 h. Plasma glucose and fructose concentrations were determined before and 4 h after each of the seven feedings. Early postpartal feeding of fructose suppressed plasma glucose (approximately 50%), with a reciprocal rise in plasma fructose. Irrespective of treatment, plasma glucose concentrations did not stabilize (approximately 100 mg/dL) until 17 to 24 h after birth. After 24 h, lactose supplements increased concentrations of plasma glucose 4 h after supplementation (169.7 +/- 8.2 mg/dL), compared with those in calves that did not receive the additional lactose. After 24 h, fructose supplements did not affect plasma glucose, but plasma fructose concentrations increased (82.6 +/- 12.4 mg/dL) 4 h after administration. The response to fructose supplements declined by 11.4 mg x dL(-1) x d(-1). Fructose was not detected in the plasma of control or lactose-treated calves after 17 h after birth. Calves that received fructose supplements had rectal temperatures 8 and 10 h after birth that were higher than those of the other calves. The mechanisms of sugar metabolism change quickly following birth. Oral sugar supplements increase the total plasma sugar concentrations of treated calves. PMID- 9734873 TI - Rapid communication: molecular cloning of the porcine corticotropin-releasing factor gene. PMID- 9734875 TI - Rapid communication: polymorphic (GT)n microsatellite in the bovine somatotropin receptor gene promoter. PMID- 9734874 TI - Rapid communication: nucleotide sequence of porcine and ovine tRNA(Lys) and ATPase8 mitochondrial genes. PMID- 9734876 TI - Rapid communication: localization of the porcine carboxypeptidase-E gene by linkage analysis further extends the region of synteny between human chromosome 4 and porcine chromosome 8. PMID- 9734877 TI - First lessons from the "Bristol case". PMID- 9734878 TI - Striking a balance in maternal immune response to infection. PMID- 9734879 TI - Disparate amoebae. PMID- 9734880 TI - Fading reveries: repressed-memory madness in the UK. PMID- 9734881 TI - Should A2 kidneys be transplanted into B or O recipients? PMID- 9734882 TI - Malthus, a prophet without honour. PMID- 9734883 TI - Adjuvant portal-vein infusion of fluorouracil and heparin in colorectal cancer: a randomised trial. European Organisation for Research and Treatment of Cancer Gastrointestinal Tract Cancer Cooperative Group, the Gruppo Interdisciplinare Valutazione Interventi in Oncologia, and the Japanese Foundation for Cancer Research. AB - BACKGROUND: There is conflicting evidence on the efficacy of regional adjuvant chemotherapy, via portal-vein infusion (PVI), after resection of colorectal cancer. We undertook a randomised controlled multicentre trial to investigate the efficacy of PVI (500 mg/m2 fluorouracil plus 5000 IU heparin daily for 7 days). METHODS: 1235 of about 1500 potentially eligible patients were randomly assigned surgery plus PVI or surgery alone (control). The patients were followed up for a median of 63 months, with yearly screening for recurrent disease. The primary endpoint was survival; analyses were by intention to treat. FINDINGS: 619 patients in the control group and 616 in the PVI group met eligibility criteria. 164 (26%) control-group patients and 173 (28%) PVI-group patients died. 5-year survival did not differ significantly between the groups (73 vs 72%; 95% Cl for difference -6 to 4). The control and PVI groups were also similar in terms of disease-free survival at 5 years (67 vs 65%) and the number of patients with liver metastases (79 vs 77%). INTERPRETATION: PVI of fluorouracil, at a dose of 500 mg/m2 for 7 days, cannot be recommended as the sole adjuvant treatment for high-risk colorectal cancer after complete surgical excision. However, these results cannot eliminate a small benefit when PVI is used at a higher dosage or in combination with mitomycin. PMID- 9734884 TI - Long-lasting recovery in CD4 T-cell function and viral-load reduction after highly active antiretroviral therapy in advanced HIV-1 disease. AB - BACKGROUND: Highly active antiretroviral therapy (HAART) decreases viral load and increases CD4 T-cell counts in patients with advanced HIV-1 infection. Whether HAART can improve CD4 T-cell function, and the biological characteristics affecting immune reconstitution, remain unclear. We undertook an open prospective pilot study to address these issues. Both treatment-naive and previously treated patients were included. METHODS: 20 patients (seven naive, 13 previously treated) were treated with one protease inhibitor and two reverse-transcriptase inhibitors and followed up for 12 months. We measured CD4-cell proliferation in response to cytomegalovirus and tuberculin antigens and counted subsets of CD4 cells at baseline and months 1, 3, 6, 9, and 12. Patients who had no antigen-specific reactivity at baseline but developed it while receiving HAART were classified as immunological responders. FINDINGS: Four patients had antigen-specific reactivity at baseline compared with 14 at month 12 (p <0.001). Between month 3 and month 12 viral load fell by a median of 1.5 log copies/mL from baseline (4.6 log copies/mL) and CD4-cell count increased by a median of 63/microL (from 93/microL). Ten patients (six of seven naive, four of 13 previously treated) were immunological responders. They differed significantly from the ten non-responders in that their viral-load reduction was sustained for 12 months, the increase in CD4 count was greater, and they showed an early increase in memory CD4 T cells with an increase of naive T cells. INTERPRETATION: HAART can induce sustained recovery of CD4 T-cell reactivity against opportunistic pathogens in severely immunosuppressed patients. This recovery depends not on baseline values but on the amplitude and duration of viral-load reduction and the increase of memory CD4 T cells. PMID- 9734885 TI - Natural history of scoliosis in spastic cerebral palsy. AB - BACKGROUND: Although the frequent occurrence of scoliosis in patients who have spastic cerebral palsy is well known and surgical treatment has often been recommended for these patients, little is known about the natural history of scoliosis in this population. We aimed to clarify the natural history of scoliosis from childhood through to adulthood and provide objective data on proper surgical indications for such patients. METHODS: The participants were 37 institutionalised patients with severe spastic cerebral palsy and scoliosis. All the participants had a series of radiographs taken, starting at a mean age of 7.8 years; they were followed up for an average of 17.3 years. We retrospectively reviewed radiographs and assessed the effect of five factors on progression of scoliosis: sex, degree of spasticity, initial physical capability, pattern of spinal curve, and location of curve. FINDINGS: Scoliosis usually started before the age of 10 years and progressed rapidly during the growth period. In many cases, even after growth had ended, continuous progression was seen. The mean magnitude of the curves at final examination was 55 degrees (Cobb angle). In 11 (85%) of 13 patients who had a spinal curve of more than 40 degrees before age 15 years, the scoliosis progressed to more than 60 degrees by the time of the final examination. Meanwhile, in only three (13%) of 24 patients who had a curve of less than 40 degrees at age 15 years, did the scoliosis progress to more than 60 degrees. Severe scoliosis (> or = 60 degrees) developed predominantly in those who had total body involvement (67%), were bedridden (100%), or had throacolumbar curves (57%). INTERPRETATION: The risk factors for progression of scoliosis in spastic cerebral palsy are: having a spinal curve of 40 degrees before age 15 years; having total body involvement; being bedridden; and having a thoracolumbar curve. Patients with these risk factors might benefit from early surgical intervention to prevent progression to severe scoliosis. PMID- 9734886 TI - Plasma bradykinin in angio-oedema. AB - BACKGROUND: Bradykinin is believed to be the main mediator of symptoms in hereditary (HA) and acquired (AA) angio-oedema due to C1 esterase inhibitor deficiency, as well as in angio-oedema that complicates treatment with inhibitors of angiotensin-converting enzyme (ACE). Difficulties in the measurement of kinin concentrations, however, have so far precluded the demonstration of an incontrovertible change in plasma bradykinin concentrations in these disorders. By developing a reliable assay we have been able to follow bradykinin concentrations during attacks and during remission in HA and in AA, and also in a patient treated with an ACE-inhibitor. METHODS: Liquid-phase extraction, high performance liquid chromatography, and RIA were used for specific measurement of plasma bradykinin concentrations in 22 patients with HA and in 22 healthy volunteers of similar age and sex distribution. Four patients with AA and one hypertensive patient treated with the ACE inhibitor captopril were also studied. FINDINGS: Among the healthy volunteers plasma bradykinin concentration was inversely proportional to age. The geometric mean plasma bradykinin concentration in the healthy volunteers was 2.2 fmol/mL (SD 2.2), compared with 3.9 fmol/mL (3.7) among patients with HA during remission (p=0.095). Bradykinin was also high in the patients with AA (10.4 fmol/mL [1.6]). During acute attacks of oedema, in both HA and AA, plasma bradykinin rose to two to 12 times the upper limit of normal. Infusion of C1-esterase inhibitor (the deficient factor in both HA and AA) immediately lowered bradykinin concentrations. In the patient receiving the ACE-inhibitor captopril, bradykinin concentration was very high at 47 fmol/mL during an acute attack of angio-oedema, but normal at 3.2 fmol/mL in remission after withdrawal of the drug. INTERPRETATION: A sensitive method for measurement of plasma bradykinin provided the means to show that concentrations of this peptide decrease with age in healthy people. Although the differences between patients in remission and healthy controls did not reach statistical significance, there were substantial rises in bradykinin during acute attacks of hereditary, acquired, or captopril-induced angio-oedema. PMID- 9734887 TI - Community outbreak of psittacosis in a rural Australian town. AB - BACKGROUND: Health authorities in Victoria, Australia were notified of three men from a rural town with atypical pneumonia, admitted to hospital over 8 days. Initial serological testing suggested Chlamydia psittaci as the cause. We did a case-control study to find risk factors for psittacosis. METHODS: We searched for cases of pneumonia or severe flu-like illness through family physicians and the regional hospital. We selected three controls per case from the region's electoral roll. We collected blood for serological tests and administered questionnaires to all cases and controls. FINDINGS: We found 16 cases of psittacosis and one died. Most cases were clustered within a small geographical area, with a median age of 58 years (range 23-76), 15 (94%) of whom were male. Keeping, handling, or feeding domestic or wild birds was not associated with illness. Cases spent a median of 17.5 h per week in their garden, compared with a median of 5.2 h for controls (p=0.04) and were more likely to have mowed lawns during the 3 weeks before onset of illness than controls (odds ratio 4.81 [95% CI 1.08-33.37]). INTERPRETATION: We showed that psittacosis outbreaks are not limited to direct contact with birds and pose new challenges for disease control. Modifications may be needed to work outdoors to decrease the risk of psittacosis. PMID- 9734888 TI - Postpartum coma. PMID- 9734889 TI - Inhaled nitric oxide and heparin for infantile primary pulmonary hypertension. PMID- 9734890 TI - Prope tolerance, perioperative campath 1H, and low-dose cyclosporin monotherapy in renal allograft recipients. PMID- 9734891 TI - Efficient and persistent gene transfer of AAV-CFTR in maxillary sinus. PMID- 9734892 TI - Intraocular pressure and baroreflex failure. PMID- 9734894 TI - Latent foamy and simian retroviruses in healthy African green monkeys used in biomedical research. PMID- 9734893 TI - Wernicke's encephalopathy due to self starvation in a child. PMID- 9734895 TI - Bartonella henselae isolated from cats in Zimbabwe. PMID- 9734896 TI - Clothing protection factor of a replica England football shirt. PMID- 9734897 TI - Evidence against "Bristol-case" doctors found proven. PMID- 9734898 TI - Gene therapy applied to treatment of HIV-1 infection. PMID- 9734900 TI - Shortages distort the social fabric of Iraq. PMID- 9734899 TI - Tackling the real culprits in Crohn's disease. PMID- 9734901 TI - Canada finally launches national HIV/AIDS strategy. PMID- 9734902 TI - Israel trips on the torturous route to security. PMID- 9734903 TI - Atopic dermatitis. PMID- 9734904 TI - Powerful placebo: the dark side of the randomised controlled trial. PMID- 9734905 TI - Medical research and the popular media. PMID- 9734906 TI - This is what the game is about. PMID- 9734907 TI - Gynaecology, forced sterilisation, and asylum in the USA. PMID- 9734908 TI - Monoclonal antibody to TNFalpha in septic shock. PMID- 9734909 TI - Nitrates for myocardial infarction. PMID- 9734910 TI - Nitrates for myocardial infarction. PMID- 9734911 TI - High-dose chemotherapy in metastatic breast cancer. PMID- 9734912 TI - Symptomless colonisation by Clostridium difficile and risk of diarrhoea. PMID- 9734913 TI - Is digoxin a designer oestrogen? PMID- 9734914 TI - Vitamin K supplementation in patients on continuous ambulatory peritoneal dialysis. PMID- 9734915 TI - Abnormal fat distribution and use of protease inhibitors. PMID- 9734916 TI - Abnormal fat distribution and use of protease inhibitors. PMID- 9734917 TI - Abnormal fat distribution and use of protease inhibitors. PMID- 9734918 TI - Global warming and vector-borne disease. PMID- 9734919 TI - Global warming and vector-borne disease. PMID- 9734920 TI - Intravascular-catheter-related infections. PMID- 9734921 TI - Intravascular-catheter-related infections. PMID- 9734922 TI - Intravascular-catheter-related infections. PMID- 9734923 TI - Mortality data for sub-Saharan Africa. PMID- 9734924 TI - Multiple antibiotic resistance in Salmonella enterica serotype enteritidis. PMID- 9734925 TI - Ghostwriting. PMID- 9734926 TI - Ghostwriting. PMID- 9734927 TI - Cheap sell for ill health. PMID- 9734928 TI - Continuing medical education. PMID- 9734929 TI - Small normal girls. PMID- 9734930 TI - Pushing ethical pharmaceuticals direct to the public. PMID- 9734931 TI - Search for effective immunomodulating strategies against sepsis. PMID- 9734932 TI - Risks and benefits of iodine supplementation. PMID- 9734933 TI - Are HCV-infected individuals candidates for hepatitis A vaccine? PMID- 9734934 TI - Limitations of epidemiology in understanding pathogenesis of cataracts. PMID- 9734935 TI - Clinical assessment of acute coma in children. PMID- 9734936 TI - Pre-emptive bone strikes in prevention of osteoporosis. PMID- 9734937 TI - Gonorrhoea control and antimicrobial resistance. PMID- 9734938 TI - Double-blind randomised controlled trial of monoclonal antibody to human tumour necrosis factor in treatment of septic shock. NORASEPT II Study Group. AB - BACKGROUND: Despite the availability of potent antibiotics and intensive care, mortality rates from septic shock are 40-70%. We assessed the safety and efficacy of murine monoclonal antibody to human tumour necrosis factor alpha (TNF alpha MAb) in the treatment of septic shock. METHODS: In a randomised, multicentre, double-blind, placebo-controlled clinical trial in 105 hospitals in the USA and Canada, we randomly assigned 1879 patients a single infusion of 7.5 mg/kg TNF alpha MAb (n=949) or placebo (0.25% human serum albumin n=930). Our main outcome measurement was the rate of all-cause mortality at 28 days. FINDINGS: 382 (40.3%) of 948 patients who received TNF alpha MAb and 398 (42.8%) of 930 who received placebo had died at 28 days (95% CI -0.02 to 0.07, p=0.27). We found no association between therapy with TNF alpha MAb and increased rapidity in reversal of initial shock or prevention of subsequent shock. Similarly, baseline plasma interleukin-6 concentrations of more than 1000 pg/mL or detectable circulating TNF concentrations were not associated with improvement in survival after TNF alpha MAb therapy. Coagulopathy but not other organ or system failures, was significantly decreased in the TNF alpha MAb group compared with placebo (day 7, p<0.001; day 28, p=0.005). Serious adverse events were reported in 55.2% of patients given placebo and 54.1% in the TNF alpha MAb group. INTERPRETATION: We did not find an improvement in survival after septic shock with TNF alpha MAb. Therapy not solely dependent on TNF alpha blockade may be required to improve survival. PMID- 9734939 TI - Mortality differences between black and white men in the USA: contribution of income and other risk factors among men screened for the MRFIT. MRFIT Research Group. Multiple Risk Factor Intervention Trial. AB - BACKGROUND: Studies of underlying differences in adult mortality between black and white individuals in the USA have been constrained by limitations of data or small study size. We investigated the extent to which differences in socioeconomic position between black and white men contribute to differences in all-cause and cause-specific mortality. METHODS: 361,662 men were screened for the Multiple Risk Factor Intervention Trial between 1973 and 1975, in 22 sites. Median family income of households by zipcode (postal) area of residence was available for 20,224 black and 300,685 white men as well as data on age, cigarette smoking, blood pressure, serum cholesterol, previous heart attack, and treatment for diabetes. We classified deaths during 16 years of follow-up into specific causes and compared differences in death rates between black men and white men, before and after adjustment for differences in income and other risk factors. FINDINGS: Age-adjusted relative risk of death (black vs white) was 1.47 (95% CI 1.42-1.53). Adjustment for diastolic blood pressure, serum cholesterol, cigarette smoking, medication for diabetes, and previous admission to hospital for heart attack decreased the relative risk to 1.40 (1.35-1.46). Adjustment for income but not the other risk factors decreased the risk to 1.19 (1.14-1.24) and adjustment for other risk factors did not alter this estimate. For cardiovascular death, relative risk on adjustment for income was decreased from 1.36 to 1.09; for cancer from 1.47 to 1.25; and for non-cardiovascular and non-cancer deaths from 1.71 to 1.26. For some specific causes of death, including prostate cancer, myeloma, and hypertensive heart disease, the higher death rates among black men did not seem to reflect differences in income. Rates of death for suicide and melanoma were lower among black than white men, as were those for coronary heart disease after adjustment for income. INTERPRETATION: Socioeconomic position is the major contributor to differences in death rates between black and white men. Differentials in mortality from some specific causes do not simply reflect differences in income, however, and more detailed investigations are needed of how differences are influenced by environmental exposures, lifetime socioeconomic conditions, lifestyle, racism, and other sociocultural and biological factors. PMID- 9734940 TI - Randomised trial of growth hormone in short normal girls. AB - BACKGROUND: There are few data on the long-term outcome of growth-hormone treatment in short normal children. We assessed the impact of growth-hormone treatment on pubertal development and near-final height in girls. METHODS: In a randomised controlled trial, we studied ten girls, with a mean age of 8.07 years and height 2 SDs or more below the mean for their age, and eight short untreated controls matched for age, and 20 short untreated girls who did not give consent for randomisation. The girls received either 30 IU/m2 somatropin per week as daily subcutaneous injections or no treatment. We assessed pubertal staging and height gain every 6 months. FINDINGS: Eight treated girls completed a mean of 6.2 years' therapy. By a mean age of 16.4 years, their mean height SD score had changed significantly from -2.42 to -1.14 (p=0.008) and they were, on average, 7.5 cm taller than the girls in the control group (height SD scores did not change significantly from -2.55) and 6.0 cm taller than the non-consent group. The timing of each pubertal stage, and the age and amplitude of peak height velocity were similar for all groups. INTERPRETATION: Growth-hormone therapy effectively increased height SD score among short normal girls started on treatment in early to mid childhood, with no untoward effect on pubertal progression. PMID- 9734941 TI - Endothelial-cell permeability and protein kinase C in pre-eclampsia. AB - BACKGROUND: Oedema and vascular leakage play a part in the pathogenesis of pre eclampsia. We tested the hypothesis that serum from pre-eclamptic patients increases endothelial-cell permeability and examined possible signal-transduction pathways. METHODS: We studied eight patients with pre-eclampsia, eight normotensive pregnant women, eight non-pregnant women, five pregnant patients with pre-existing hypertension, and four hypertensive non-pregnant women. Cultured human umbilical-vein endothelial-cell monolayers were used and permeability was measured by albumin flux. The part played by protein kinase C (PKC) signalling was examined by down-regulation with phorbol ester and with the inhibitors Goe 6976 and staurosporine. PKC isoforms were assessed by western blot and confocal microscopy. Antisense oligodesoxynucleotides (ODN) were used to test for specific PKC isoforms. FINDINGS: Serum from pre-eclamptic women increased endothelial permeability significantly (by 100%, p<0.01). The change in permeability decreased rapidly after delivery. Serum from normotensive pregnant women and non-pregnant women had no effect. Permeability was not influenced by serum from patients with essential hypertension or pregnant patients with pre existing hypertension. Serum from pre-eclamptic patients induced a translocation of PKC isoforms alpha and epsilon within the cells. Goe 6976 and staurosporine (10(-8) mol/L) inhibited the increase in permeability induced by serum from pre eclamptic patients. Down-regulation of PKC alpha and, to a lesser extent, PKC epsilon by antisense ODN also inhibited the pre-eclampsia-induced permeability increase. INTERPRETATION: Serum from pre-eclamptic patients contains a factor or factors that increase endothelial-cell permeability. The effect of pre-eclamptic serum may be mediated by PKC alpha and epsilon. PMID- 9734942 TI - Anti-inflammatory cytokine profile and mortality in febrile patients. AB - BACKGROUND: An anti-inflammatory cytokine profile on whole-blood stimulation in vitro is associated with fatal outcome of meningococcal disease. We investigated whether an anti-inflammatory cytokine profile in the circulation is associated with adverse outcome in other infectious diseases. METHODS: We enrolled 464 consecutive patients (272 men, 192 women) who presented to hospital with fever (> or = 38.2 degrees C). On admission we measured plasma interleukin 10 (IL-10) and tumour necrosis factor alpha (TNF alpha), and collected clinical and microbiological data on the febrile illness, then followed up all patients for clinical outcome. FINDINGS: In at least 399 of the 464 patients fever was caused by infection. 33 patients died after a median hospital stay of 11 days (interquartile range 3-20). Concentrations of IL-10 were significantly higher in non-survivors (median 169 pg/mL [IQR 83-530]) than in survivors (median 88 pg/mL [42-235], p=0.042). When dichotomised around the median, the mortality risk was two times higher in patients who had high concentrations of IL-10 than in those with low concentrations (relative risk 2.39 [95% CI 1.07-5.33]), in patients with low and high concentrations of TNF alpha. In the 406 patients without haemodynamic deterioration in the first 24 h, IL-10 was higher and TNF alpha lower in patients who died than in those who survived. The ratio of IL-10 to TNF alpha was higher in non-survivors (median 6.9 [3.0-21.0]) than in survivors (median 3.9 [2.0-7.0], p=0.040). This ratio was highest in patients who died without underlying disease (median 21.5 [5.0-25.0]). Age, sex, and duration of fever before admission did not explain the differences in IL-10 and TNF alpha. INTERPRETATION: An anti-inflammatory cytokine profile of a high ratio of IL-10 to TNF alpha is associated with fatal outcome in febrile patients with community acquired infection. Our findings caution against a widespread use of proinflammatory cytokine inhibition in patients with sepsis. PMID- 9734943 TI - A farmer with a lump in his throat. PMID- 9734944 TI - Inflammatory bowel disease and autism. PMID- 9734945 TI - Parvovirus B19 and acute hepatitis in adults. PMID- 9734946 TI - Amyloid beta protein in skin of patients with amyotrophic lateral sclerosis. PMID- 9734947 TI - Evidence of disturbed meiosis in a man referred for intracytoplasmic sperm injection. PMID- 9734948 TI - Fertility drugs and malignant germ-cell tumour of ovary in pregnancy. PMID- 9734949 TI - Lamotrigine and topiramate may be a useful combination. PMID- 9734950 TI - Granulocytosis causing sickle-cell crisis. PMID- 9734951 TI - Insulin vasodilatation and the "arginine paradox". PMID- 9734952 TI - Silver in sugar particles and systemic argyria. PMID- 9734953 TI - Vitamin E may reduce prostate-cancer incidence. PMID- 9734954 TI - Experts plan strategy to tackle mother-to-child HIV-1 transmission. PMID- 9734955 TI - Easing the way to safer sex. PMID- 9734956 TI - A new financial injection for the National Health Service. PMID- 9734957 TI - Juvenile chronic arthritis. PMID- 9734959 TI - Genetically determined failure of activation-induced apoptosis of autoreactive T cells as a cause of multiple sclerosis. AB - I postulate that multiple sclerosis is an autoimmune disease that involves genetically determined failure of activation-induced apoptosis of autoreactive T cells in the central nervous system. Activation of central-nervous-system reactive T cells in peripheral lymphoid organs by exposure to crossreacting antigens or superantigens derived from common infectious agents may trigger attacks of multiple sclerosis. In normal individuals these activated T cells are deleted by activation-induced apoptosis, but in individuals predisposed to multiple sclerosis they survive, proliferate, and damage the central nervous system. The clinical course of multiple sclerosis may vary according to the antigens in the central nervous system being targeted: targeting of myelin antigens leads to a relapsing-remitting course of clinical recovery due to remyelination or other mechanisms; targeting of axonal antigens leads to a progressive course from onset because axonal regeneration is limited in the central nervous system. This hypothesis can account for many characteristics of multiple sclerosis and has predictions that can be tested. PMID- 9734958 TI - The Nuremberg Code: Hippocratic ethics and human rights. PMID- 9734960 TI - Physicians as caretakers and collaborators. PMID- 9734961 TI - Mozambique: investing in peace. PMID- 9734962 TI - Laparoscopy versus endoscopy for bile-duct stones. PMID- 9734963 TI - Laparoscopy versus endoscopy for bile-duct stones. PMID- 9734964 TI - Laparoscopy versus endoscopy for bile-duct stones. PMID- 9734965 TI - Laparoscopy versus endoscopy for bile-duct stones. PMID- 9734966 TI - High-dose chemotherapy in metastatic breast cancer. PMID- 9734967 TI - Protein-C concentrate for meningococcal purpura fulminans. PMID- 9734968 TI - Protein-C concentrate for meningococcal purpura fulminans. PMID- 9734969 TI - Protein-C concentrate for meningococcal purpura fulminans. PMID- 9734970 TI - Protein-C concentrate for meningococcal purpura fulminans. PMID- 9734971 TI - Cost-effectiveness estimates of the Mwanza sexually transmitted diseases intervention. PMID- 9734972 TI - Losartan and renal transplantation. PMID- 9734973 TI - Valvular heart disease and Chinese-herb nephropathy. PMID- 9734974 TI - Valvular heart disease and Chinese-herb nephropathy. PMID- 9734975 TI - Maternal mortality. PMID- 9734976 TI - Lice buried under the ashes of Herculaneum. PMID- 9734977 TI - Mucosal Immunity in HIV Infection. Proceedings of the International Symposium. Berlin, Germany, June 6-7, 1997. PMID- 9734978 TI - Cattle TB Crisis? Human implications. PMID- 9734979 TI - [Zonisamide:its placental transport, biological half-life in the newborn, and transport to mother's milk--a study of a case of an infant born of a mother who had been treated with zonisamide alone during pregnancy]. PMID- 9734980 TI - [Changes in serum anti-Helicobacter pylori antibody level in severely handicapped children and adults after one year of institutionalization]. PMID- 9734981 TI - Papillary cystic neoplasm of the pancreas: radiological findings. AB - We report a series of 10 papillary cystic neoplasms of the pancreas evaluated in our institution. The lesions are analyzed in retrospect to define the existence of eventual specific imaging patterns as well as to point out the existing problems of differential diagnosis versus other pancreatic tumors. PMID- 9734982 TI - Cloning and sequence analysis of the Candida tropicalis URA3 gene encoding orotidine-5'-phosphate decarboxylase. AB - An integrative transformation system was established for a phenol-utilizing strain of Candida tropicalis M4. The system is based on an auxotrophic mutant host of C. tropicalis U-6 that is defective in orotidine-5'-phosphate decarboxylase (ODCase). As a selectable marker, we isolated and characterized the C. tropicalis URA3 gene, which codes for ODCase. The gene was cloned by complementation of the ura3 mutation of Sachharomyces cerevisiae SHY-3 and the pyrF mutation of Escherichia coli. The C. tropicalis U-6 was transformed by plasmid containing the C. tropicalis URA3 gene at a frequency of 1 to 10 transformants per microgram of plasmid DNA. When the URA3 gene was expressed in E. coli minicells, a 30-kDa protein was identified. Nucleotide sequence analysis revealed the presence of an open reading frame, encoding a protein of 268 amino acids with a calculated molecular mass of 29.7 kDa. The nucleotide sequence of URA3 gene and its deduced amino acid sequence showed significant homology to those of the ODCase of other fungal species. PMID- 9734983 TI - Urological Research Society annual scientific meeting. London, United Kingdom, 9 January 1998. Abstracts. PMID- 9734984 TI - A focus on implementing nursing vocabularies. PMID- 9734985 TI - Veterinary practice management. Boom or bust: the impact of changing demographics on today's practice. PMID- 9734986 TI - Urogynecology fellowship training program directory. PMID- 9734987 TI - Current awareness in geriatric psychiatry. PMID- 9734988 TI - Net to help catch NZ serum forgers. PMID- 9734989 TI - The Delphic boat or what the genomic texts tell us. PMID- 9734990 TI - Drugs for pediatric emergencies. Committee on Drugs, Committee on Drugs, 1996 to 1997, Liaison Representatives, and AAP Section Liaisons. AB - This statement provides current recommendations about the use of emergency drugs for acute pediatric problems that require pharmacologic intervention. At each clinical setting, physicians and other providers should evaluate drug, equipment, and training needs. The information provided here is not all-inclusive and is not intended to be appropriate to every health care setting. When possible, dosage recommendations are consistent with those in standard references, such as the Advanced Pediatric Life Support (APLS) and Pediatric Advanced Life Support (PALS) textbooks. Additional guidance is available in the manual Emergency Medical Services for Children: The Role of the Primary Care Provider, published by the American Academy of Pediatrics, as well as in the PALS and APLS textbooks. PMID- 9734992 TI - The politics of patent legislation in biotechnology: an international review. AB - The realization of the economic value of the genetic resources has prompted an international debate about property rights to genetic resources. The international debate pertaining to patenting of genetic material is the main theme of this chapter. As a backdrop for the international debate, the chapter starts out with a summary of the main events and arguments in the expanding scope of patent legislation in biotechnological inventions. Summing up, the new biotechnologies represent a tool which meets the legal requirements for patenting biological material. From the industry's point of view, biotechnology also necessitates patenting. On the negative side, defending a patent is often a long and costly business, and the trend is that patenting will mainly benefit the bigger and stronger companies and thus weaken public control over the rapid developments in biotechnology. A central argument in the chapter is that without sophisticated biotechnological tools, trained scientists, and adequate infrastructure, patenting is, as yet, hardly a viable solution for the majority of developing countries. Gene-rich developing countries fear that developments in patent legislation will pave the way for increased Northern control over Third World natural resources. The International Convention on Biological Diversity goes some way in making amends to this situation, but the gene-poor, least developed countries may still have reason to fear that they will lose access to breeding material. In a long-term perspective, the implications may be detrimental for resource conservation in developing countries. In conclusion, the patent question seems to remain unresolved and may still be one of the most likely stumbling blocks for future ratifications and implementation of the Biodiversity Convention. PMID- 9734991 TI - Solid-phase technology: magnetic heads to improve nucleic acid detection and analysis. PMID- 9734993 TI - Combinatorial approaches to polyketide biosynthesis. AB - Polyketides are a large and structurally diverse family of natural products based on chains of carboxylic acid units. The polyketide synthases that make aromatic polyketides have already been used to generate small combinatorial libraries, by expressing individual genes from different biosynthetic pathways together, so that the enzymes they encode can interact to make novel products. Recent work has shown how to choose these individual components to increase the chances of obtaining such hybrid aromatic compounds. In other polyketide synthases which synthesise complex reduced polyketides, the constituent enzymes are actually linked as domains in a giant multienzyme complex along which the growing polyketide chain is passed. A combinatorial approach here therefore requires the fusing together of individual enzymatic domains from several such synthases in as many productive ways as can be devised, so that the enzyme assembly line produces a library of altered products. A key recent advance has been to demonstrate that such genuinely hybrid enzymes do work as predicted, for example a broad specificity enzyme that recruits the chain starter unit for an antiparasitic compound has been grafted onto a synthase that makes antimicrobial macrolides. PMID- 9734994 TI - Bibliography. Current world literature. Model systems. PMID- 9734995 TI - Bibliography. Current world literature. Biopolymers. PMID- 9734996 TI - Our contribution to the public fear of cancer. PMID- 9734997 TI - Ethical questions on the use of magnetic field reports. PMID- 9734999 TI - Healing venom. PMID- 9734998 TI - The importance of protocol design and data reporting to research on endocrine disruption. PMID- 9735000 TI - Bird flu vaccine. PMID- 9735001 TI - The state of the science on endocrine disruptors. PMID- 9735002 TI - Landfills: is wetter better? PMID- 9735003 TI - [The Giornale Italiano di Cardiologia and the ANMCO symbol. Associazione Nazionale Medici Cardiologi Ospedalieri]. PMID- 9735004 TI - Recombination-dependent mutation in Escherichia coli occurs in stationary phase. PMID- 9735005 TI - 2nd Annual World Congress on Allied Health. Telford, England, July 23, 1997. Proceedings and abstracts. PMID- 9735006 TI - 24th Annual meeting of the Japanese Society for Ultrastructural Cutaneous Biology. Hirosaki, Japan, October 17-18, 1997. Abstracts. PMID- 9735007 TI - Proceedings of a symposium to mark the retirement of Professor Donald L. Lee. University of Leeds, September 1996. PMID- 9735008 TI - Report of an international meeting on rubella vaccines and vaccination, 9 August 1993, Glasgow, United Kingdom. PMID- 9735009 TI - A new method to correlate acoustic spectroscopic microscopy (30 MHz) and light microscopy. AB - A powerful new method is used to investigate the correlation between light microscopic and acoustic properties of biological tissues. Specimens of liver were sectioned into successive slices, 250 micrometers and 10 micrometers thick. The thick sections were investigated acoustically, the thin sections by means of light microscopy. Markers that could be detected and located, both optically and acoustically, were used to find and reconstruct corresponding regions in the acoustic and optical sections (2.5 x 2.5 mm). Parameter images were reconstructed from the sections investigated acoustically. The acoustic parameters were attenuation at 30 MHz, the slope of the attenuation spectrum (between 10 and 50 MHz), backscattering at 30 MHz, the slope of the backscattering spectrum (between 10 and 50 MHz) and the local ultrasound velocity. Acoustic images were obtained in the frequency range from 10 to 50 MHz, yielding a lateral resolution of about 50 micrometers. The sections for light microscopy were stained according to the Goldner trichrome staining technique. The histological composition was determined quantitatively, using digital image segmentation techniques. The percentage of collagen-rich fibrous tissue, luminal structure and interstitial spaces, and the number of nuclei were calculated for regions of 250 x 250 micrometers. These histological features were correlated with the acoustic parameters obtained from the corresponding regions in adjacent sections. It was thus possible to find the histological components responsible for acoustic parameters. PMID- 9735010 TI - Pirarubicin-induced myocardial damage in elderly patients with non-Hodgkin's lymphoma. AB - 123I-metaiodobenzylguanidine (MIBG) myocardial single photon emission computed tomography (SPECT), 123I-beta-methyliodophenyl pentadecanoic acid (BMIPP) myocardial SPECT, and holter ECG recording were performed in patients with non Hodgkin's lymphoma who underwent chemotherapy including pirarubicin (THP), in an attempt at early detection of cardiac toxicity from THP. Twenty-six patients with untreated non-Hodgkin's lymphoma who received THP-COPBLM therapy were studied. For THP-COPBLM therapy. THP was administered at a dose of 40 mg/m2 every 21 days and the total dose was 250 mg/m2 on average (40 approximately 400 mg/m2). 1) The washout rate (WR) correlated with the total THP dose, and was considered to be a useful index of cardiac sympathetic nervous dysfunction. 2) The left ventricular ejection fraction (LVEF) correlated negatively with the total dose of THP. 3) The total dose of THP showed a correlated positively with the extent score and severity score determined by BMIPP. 4) The WR correlated with the frequency of premature ventricular contraction. Animal studies have indicated that THP has less cardiac toxicity than doxorubicin, but the present study showed that cardiac toxicity occurred at a total THP dose of about 360 mg/m2 in elderly patients. Accordingly, when THP is used to treat elderly patients, multimodal evaluation of cardiac is necessary to detect cardiotoxicity and to determine the optimal dosage. PMID- 9735011 TI - Reflections on the origin of the genetic code: a hypothesis. AB - The origin of the organisation of the genetic code reflects both the biosynthetic relationships between amino acids and the physicochemical interactions between these and anticodons; moreover, these two forces do not act independently. It must therefore be explained why it is simultaneously true that the anticodons of product amino acids were assigned prior to their biosynthetic appearance and that physicochemical correlations must exist between anticodons and amino acids. This gives rise to difficulties of interpretation, even of a more general nature, in the theories that have been proposed to explain the origin of the genetic code; a hypothesis is thus presented in this manuscript. In particular, the hypothesis suggests that RNA hairpin structures, the ancestors of tRNAs, housing anticodon like nucleotides in the stem were charged with precursor amino acids. As the precursor amino acids gradually developed into product amino acids, a coevolution came into being between the development of amino acids and that of anticodons, with a concomitant formation of the complete tRNA molecule thought direct duplication of the hairpin structures. All this led to the definition of the genetic code organisation. Furthermore, this made it possible for the evolving anticodons to select the emerging product amino acids, which the hypothesis therefore considers to be unspecified in the initial phase of genetic code evolution but which were selected also because of their ability to interact with anticodons. In this way the main obstacle to interpretation is removed. Fossils of these events can now be observed in some amino acid-modified nucleosides specifically located in the tRNA anticodon loops. This is all presented in the framework of a general discussion of the ideas and data in favour of a late origin of the genetic code, as opposed to an early origin in the context of the theories proposed to explain the origin of the genetic code organisation. PMID- 9735012 TI - Buffering: a possible passive-homeostasis role for redundant DNA. AB - A new general role for redundant (apparently non-coding) DNA is speculated: buffering the effect of fluctuations in intra-cellular solute composition on chromatin condensation state in its condensed form and on binding of DNA-tropic proteins and other ligands in its decondensed form. The "buffering" DNA performing such passive-homeostasis (energy-independent) function, can provide selective advantage in ecological niches with a low energy supply. PMID- 9735013 TI - [Human immunodeficiency virus. New aspects of pathogenesis]. PMID- 9735014 TI - Quantitation of fumonisins in corn by HPLC with o-phthalaldehyde postcolumn derivatization and their identification by LC/MS. AB - Fumonisins B1 (FB1) and B2 (FB2) were isocratically separated on a fluorocarbon column without using an ion pair reagent and nonvolatile buffer during the HPLC and were detected by an o-phthalaldehyde postcolumn derivatization system using a fluorescence detector. The minimum detectable concentrations of FB1 and FB2 in corn by this system were 0.01 micrograms/g and 0.01 micrograms/g, respectively. The separated fumonisins were further identified by a directly interfaced ion trap MS using electrospray ionization. FB1 and FB2 in naturally contaminated corn were identified in the selective ion monitoring mode at concentrations of 3.75 micrograms/g and 1.44 micrograms/g, respectively. PMID- 9735015 TI - Inhibition of aflatoxin production of Aspergillus parasiticus NRRL 2999 by Bacillus pumilus. AB - Six isolates of Bacillus pumilus were tested for their ability to inhibit aflatoxin production of Aspergillus parasiticus NRRL 2999 in yeast extract sucrose (YES) broth. Aflatoxin production was inhibited in both simultaneous and deferred antagonism assays, suggesting that the inhibitory activity was due to extracellular metabolites(s) produced in cell-free supernatant fluids of cultured broth. The inhibition was not due to organic acids or hydrogen peroxide produced by B. pumilus since the inhibitory activity was not lost after pH adjustment or treatment of supernatant fluids with catalase. A range of media tested for the production of inhibitory metabolite(s) in supernatant fluids showed that all media supported bacterial growth and production of the metabolite(s). The metabolite(s) were produced over a wide range of temperature (25 to 37 degrees C) and pH (4 to 9) of growth of B. pumilus. They were stable over a wide range of pH (4 to 10) and were not inactivated after autoclaving at 121 degrees C for 30 minutes. PMID- 9735016 TI - Strategies against malaria. Eradication or control? Proceedings of a conference. Annecy, France, April 17-26, 1996. PMID- 9735017 TI - Stable coronary artery disease. Ongoing management, patient management, patient education, and medical interventions. Institute for Clinical Systems Integration. PMID- 9735018 TI - Optical mark recognition. Tallying information from filled-in 'bubbles'. PMID- 9735019 TI - The 7th European Placenta Group Meeting, Vigso, Denmark, 13-17 December 1997. PMID- 9735020 TI - All our dreams our sold. PMID- 9735021 TI - Geneticists debate eugenics and China' infant health law. PMID- 9735022 TI - Doubled genes may explain fish diversity. PMID- 9735023 TI - A second private genome project. PMID- 9735024 TI - DNA chips survey an entire genome. PMID- 9735025 TI - How embryos may avoid immune attack. PMID- 9735026 TI - Institute copes with genetic hot potato. PMID- 9735027 TI - How the genome readies itself for evolution. PMID- 9735028 TI - Rx for WHO. PMID- 9735029 TI - A vision of the pore. PMID- 9735030 TI - Memories are made of this. PMID- 9735031 TI - Sustained delivery of proteins for novel therapeutic agents. PMID- 9735033 TI - Cryopreservation. A Frankenstein experiment. PMID- 9735032 TI - Molecular markers. Tissue cork borer. PMID- 9735034 TI - C, N, and noble gas isotopes in grain size separates of presolar diamonds from Efremovka. AB - Nanometer-size presolar diamonds from the Efremovka CV3 chondrite were physically separated into several grain size fractions by ultracentrifugation. The coarsest size fraction is the most enriched in carbon-12; the others have broadly similar carbon isotopic compositions. Measurement of noble gases shows that their concentration decreases with decreasing grain size. This effect is attributed to ion implantation. Such an episode could occur in the envelope of a supernova that produced the diamonds, or in interstellar space; in either case, ions with energies above a certain threshold pass completely through the smaller diamond grains without being captured. Concentrations of nitrogen show only minor variations with grain size, indicating a different mechanism of incorporation into the diamonds. PMID- 9735035 TI - Medical school caught up in Pennsylvania hospital debacle. PMID- 9735036 TI - NIH, DuPont declare truce in mouse war. PMID- 9735037 TI - A new route to treating schizophrenia? PMID- 9735038 TI - NIH concocts a booster shot for HIV vaccines. PMID- 9735039 TI - Botanical gardens cope with bioprospecting loophole. PMID- 9735040 TI - Cell death in us and others. PMID- 9735041 TI - Beetle juice. PMID- 9735042 TI - A "humouse" project. PMID- 9735043 TI - DRD2 gene and alcoholism. PMID- 9735044 TI - Testing for Alzheimer's. PMID- 9735045 TI - Haeckel's embryos, continued. PMID- 9735046 TI - Food production, population growth, and the environment. PMID- 9735047 TI - Apoptosis. Death by crowd control. PMID- 9735048 TI - Stroke-damaged neurons may commit cellular suicide. PMID- 9735050 TI - The Bcl-2 protein family: arbiters of cell survival. AB - Bcl-2 and related cytoplasmic proteins are key regulators of apoptosis, the cell suicide program critical for development, tissue homeostasis, and protection against pathogens. Those most similar to Bcl-2 promote cell survival by inhibiting adapters needed for activation of the proteases (caspases) that dismantle the cell. More distant relatives instead promote apoptosis, apparently through mechanisms that include displacing the adapters from the pro-survival proteins. Thus, for many but not all apoptotic signals, the balance between these competing activities determines cell fate. Bcl-2 family members are essential for maintenance of major organ systems, and mutations affecting them are implicated in cancer. PMID- 9735049 TI - Is apoptosis key in Alzheimer's disease? PMID- 9735051 TI - Alabama Museum of the Health Sciences and Reynolds Historical Library, Birmingham, Alabama. PMID- 9735052 TI - Life in the 21st century - a vision for all. PMID- 9735053 TI - The EBCT can of worms. PMID- 9735054 TI - The EBCT controversy -- a sorry saga. PMID- 9735055 TI - Ethics and the sanctity of human life. PMID- 9735057 TI - Penicillium expansum growth and production of patulin in the presence of benzoic acid and its derivatives. AB - This study was conducted in order to evaluate the influence of benzoic acid and its derivatives: o-nitrobenzoic acid, o- and p-aminobenzoic acids, o hydroxybenzoic acid, acetylsalicylic acid, cinnamic acid and aldehyde, methyl and ethyl esters of benzoic acid, methyl ester of p-hydroxybenzoic acid and ethyl ester of p-aminobenzoic acid on the growth and the ability to produce patulin by Penicillium expansum. The growth of the mycelium and its ability to produce patulin in the liquid enriched Czapek medium, and the dynamics of its linear growth and sporulation, on the Sabouraud medium with addition of the above mentioned compounds were evaluated. Benzoic acid and most of its derivatives, within the studied range of concentrations, limited the growth of the microfungus, while methyl ester of benzoic acid and o-aminobenzoic acid stimulated its growth. PMID- 9735056 TI - Recent Advances in Liver Research: A Symposium in Honor of Professor James L. Boyer. New Haven, Connecticut, USA. May 8-9, 1997. Proceedings. PMID- 9735058 TI - Microbial degradation of phenol in denitrifying conditions. AB - The biodegradation of phenpol in anaerobic conditions by mixed population of bacteria in batch cultures or continuous cultures in packed bed reactor in medium with phenol as sole carbon source was effective. Phenol in concentrations up to 500 mg/l was degraded by bacteria in batch cultures (incubation temperature 30 degrees C) with increasing maximal rate without lag phase and at higher concentrations (up to 1000 mg/l) the activity of the bacteria was preceded by a lag phase lasting from 9 to 15 days. Phenol was degraded in continuous cultures with maximum efficiency (about 2500 mg/l x day) in the following conditions: incubation temperature 30 degrees C, phenol concentration in the medium of 200 mg/l and retention time of about 2 hours. Lowering of the temperature of the culture to 13 degrees C and 20 degrees C resulted in 10 and 5-fold decrease in the efficiency of the process, expressed as mg/l X day, respectively. Analysis of the composition of the bacteria among the facultatively growing Gram-negative rods showed that the incubation temperature visibly affected the species composition and domination pattern of denitrifying bacteria although their percent participation remained the same. PMID- 9735059 TI - Sri Lanka. Political violence and ethnic conflict. AB - In recent years, Sri Lanka has experienced 2 violent rebellions in which youths have played a prominent role, 1 in the majority Sinhala community and 1 in the minority Tamil community. The former was crushed, but the latter remains ongoing, with the Liberation Tigers of Tamil Eelam, who claim to represent the Tamil minority, battling the Sinhala-dominated government. Prospects for peace in the short- and medium term appear poor. These events have generated an impressive body of interdisciplinary interpretation, but several important topics have received relatively little attention. Most ongoing research is being carried out by anthropologists, historians, and political scientists, but psychological insights would offer important complementary perspectives. PMID- 9735060 TI - Ethnic conflict and the psychology of liberation in Guatemala, Peru, and Puerto Rico. AB - Ethnic identity and conflict in Guatemala, Peru, and Puerto Rico are complexly embedded within dynamic systems of class- and race-based geopolitics. Whereas overt violence and terror have permeated both Guatemalan and Peruvian societies, overt conflict has undermined Puerto Rican nationhood. Despite similarities among these 3 countries of Hispano-America, there are important particularities that inform psychological theory and practice. This article explores selected contributions of a psychology of liberation informed by indigenous psychologies and reflexive praxis. The challenges these conflicts and their consequences pose to psychologists seeking to work with populations most deeply affected by these social inequalities are analyzed. It concludes with suggestions of how psychology can move toward the development of community-based responses to psychosocial oppression that foster enhanced individual and collective development in a context of social change. PMID- 9735061 TI - Limitations of the 1990 American College of Rheumatology classification criteria in the diagnosis of vasculitis. AB - BACKGROUND: The American College of Rheumatology (ACR) established criteria to discriminate among patients with seven types of vasculitis. Although designated as "classification criteria" for research, these criteria are often used for diagnosis. OBJECTIVE: To examine the operating characteristics of the 1990 ACR classification criteria in the diagnosis of Wegener granulomatosis, giant-cell arteritis, polyarteritis nodosa, and hypersensitivity vasculitis. DESIGN: Prospective cohort study. SETTING: University medical center and Veterans Affairs medical center. PATIENTS: 198 consecutive patients referred to rheumatologists for evaluation of possible vasculitis. MEASUREMENTS: Blinded chart audits were done to classify patients according to the 1990 ACR classification criteria for Wegener granulomatosis, polyarteritis nodosa, giant-cell arteritis, and hypersensitivity vasculitis on the basis of the patients' initial presentation. Chart audits done 2 to 8 months after baseline provided the patients' final diagnoses, which were considered the gold standard, as in the development of the ACR criteria. Test operating characteristics of the ACR classification criteria were calculated according to 2 x 2 tables for the entire cohort and for only the patients with a final diagnosis of vasculitis. RESULTS: Vasculitis was diagnosed in 51 (26%) patients. Thirty-eight (75%) of 51 patients with vasculitis and 31 (21%) of 147 patients without vasculitis met ACR criteria for one or more types of vasculitis. The positive predictive values for the four vasculitides according to ACR criteria were 17% to 29% for the entire cohort and 29% to 75% for only the patients with a final diagnosis of vasculitis. CONCLUSION: The 1990 ACR classification criteria function poorly in the diagnosis of specific vasculitides. PMID- 9735062 TI - Ethnic and sex bias in primary care screening tests for alcohol use disorders. AB - BACKGROUND: The use of self-report screening tests for alcohol use disorders in the primary care setting has been advocated. OBJECTIVE: To test for ethnic and sex bias in three self-report screening tests for alcohol use disorders in a primary care population. DESIGN: Cross-sectional study with patients randomly selected from appointment lists. SETTING: University-based family practice clinic. PATIENTS: Probability sample of 1333 adult family practice patients stratified by sex and ethnicity. MEASUREMENTS: Patients completed 1) a diagnostic interview to determine the presence of a current alcohol use disorder and 2) three screening tests: the CAGE questionnaire, the Self-Administered Alcoholism Screening Test (SAAST), and the Alcohol Use Disorders Identification Test (AUDIT). RESULTS: The areas under the receiver-operating characteristic (ROC) curves for the CAGE questionnaire and the SAAST ranged from 0.61 to 0.88 and were particularly poor for African-American men and Mexican-American women. For the AUDIT, the area under the ROC curves was greater than 0.90 for each patient subgroup. The sensitivity of the CAGE questionnaire and the SAAST at standard cut points was lowest for Mexican-American women (0.21 and 0.13, respectively). Positive likelihood ratios for the AUDIT were similar to or higher than those for the other screening tests, whereas negative likelihood ratios were lowest for the AUDIT (<0.33), indicating the superiority of this test in ruling out a disorder. CONCLUSIONS: A marked inconsistency in the accuracy of common self-report screening tests for alcohol use disorders was found when these tests were used in a single clinical site with male and female family practice patients of different ethnic backgrounds. The AUDIT does not seem to be affected by ethnic and sex bias. PMID- 9735063 TI - Can inexpensive signs encourage the use of stairs? Results from a community intervention. AB - BACKGROUND: The U.S. Surgeon General advocates the accumulation of moderate intensity activity throughout the day to improve health. OBJECTIVES: To test the effectiveness of signs to encourage use of stairs instead of escalators. DESIGN: Community intervention. SETTING: Shopping center. PARTICIPANTS: 17901 shoppers. INTERVENTION: Signs promoting the health and weight-control benefits of stair use were placed beside escalators with adjacent stairs. MEASUREMENTS: The sex, age, race, weight classification, and use of stairs were observed. RESULTS: Overall, stair use increased from 4.8% to 6.9% and 7.2% with the health and weight-control signs, respectively. Younger persons increase their stair use from 4.6% to 6.0% with the health sign and 6.1% with the weight-control sign. Older persons almost doubled their stair use from 5.1% to 8.1% with the health sign and increased use to 8.7% with the weight-control sign. Differential use of stairs was observed between ethnic groups. Among white persons, stair use increased from 5.1% to 7.5 and 7.8% with the health sign and weight-control signs. Among black persons, stair use decreased from 4.1% to 3.4% with the health sign and increased to 5.0% with the weight-control sign. At baseline, lean persons used the stairs more often than overweight persons (5.4% and 3.8%, respectively). The health sign increased stair use to 7.2% among normal-weight persons and 6.3% among overweight persons; the weight-control sign prompted stair use to increase to 6.9% among persons of normal weight and to 7.6% among overweight persons. CONCLUSIONS: Simple, inexpensive interventions can increase physical activity. Research is needed to identify effective motivators to promote activity among black persons. PMID- 9735064 TI - Interferon-alpha treatment of four patients with the Churg-Strauss syndrome. AB - BACKGROUND: Interferon-alpha is reported to have a beneficial effect on patients with the idiopathic hypereosinophilic syndrome. OBJECTIVE: To study the effect of interferon-alpha on the Churg-Strauss syndrome. DESIGN: Case series. SETTING: University hospital. PATIENTS: Four patients with biopsy-proven Churg-Strauss syndrome. INTERVENTION: Interferon-alpha at dosages of 7.5 to 63 million U per week. MEASUREMENTS: Disease extent, disease activity, and blood eosinophil count in a treatment period ranging from 14 to 25 months. RESULTS: Interferon-alpha therapy led to remission of disease and a substantial reduction of the prednisolone requirement in two patients who had attained incomplete remission with cyclophosphamide or methotrexate. The third patient's condition stabilized, and the fourth patient maintained remission. During interferon-alpha therapy, the blood eosinophil count in all patients decreased in a dose-dependent manner and paralleled the extent of clinical disease and disease activity. CONCLUSIONS: Interferon-alpha may be an effective treatment for the Churg-Strauss syndrome. It seems to exert its effect primarily by producing a dose-dependent decrease in the blood eosinophil count. PMID- 9735066 TI - Cardiovascular risks to young persons on the athletic field. AB - Sudden cardiac deaths of young athletes, which are usually associated with physical exertion, continue to achieve high public visibility and generate considerable concern. Despite broad community participation in sports, such catastrophes are uncommon, occurring in about 1/200000 high school athletes per academic year. Various unsuspected congenital cardiovascular diseases are usually responsible; the most common lesions are hypertrophic cardiomyopathy and several congenital coronary artery anomalies. Selected reports suggest that arrhythmogenic right ventricular dysplasia may be a more common cause of these deaths than previously suspected. In some trained athletes with borderline increases in thickness of the left ventricular wall, mild morphologic expression of hypertrophic cardiomyopathy can often be distinguished from the physiologic consequences of athlete's heart by noninvasive clinical assessment and testing. In addition, the recognized cardiovascular risks of the athletic field are now extended to include cardiac arrest resulting from relatively modest, nonpenetrating chest blows produced by projectiles (such as baseballs) or bodily contact in the absence of underlying cardiac disease and without structural injury to the chest wall or heart. These uncommon but usually fatal events seem to result when chest impact occurs precisely during the vulnerable phase of repolarization, and they may be reduced by use of softer baseballs. Preparticipation screening for cardiovascular disease, consisting of standard history and physical examination, is customary practice for most high school and college athletes in the United States. Evidence suggests, however, that the present screening process for cardiovascular disease in high school athletes may be largely inadequate, given the content of the approved screening questionnaires (which serve as guidelines for the process) and the use of examiners with little cardiovascular training. This emphasizes the need for national standardization of preparticipation screening. The recommendations of the 26th Bethesda Conference for disqualification from competitive athletics are now a standard for management decisions when cardiovascular abnormalities are identified in trained athletes. PMID- 9735067 TI - Update in geriatrics. PMID- 9735065 TI - Exhaled nitric oxide and impaired oxygenation in cirrhotic patients before and after liver transplantation. AB - BACKGROUND: Nitric oxide may be involved in the impaired oxygenation of cirrhotic patients, a condition that improves in most patients after liver transplantation. OBJECTIVE: To compare oxygenation and nitric oxide concentrations before and after liver transplantation. DESIGN: Before-and-after observational study. SETTING: Academic medical center. PATIENTS: 18 patients with cirrhosis and no obvious cardiopulmonary disease who underwent successful orthotopic liver transplantation. INTERVENTION: Orthotopic liver transplantation. MEASUREMENTS: Blood gas analysis, measurement of exhaled nitric oxide, contrast-enhanced echocardiography, and pulmonary function tests. RESULTS: Before transplantation, the mean (+/- SD) exhaled nitric oxide concentration was higher in patients than in normal controls (13 +/- 4.9 parts per billion [ppb] compared with 5.75 +/- 1.9 ppb; P < 0.001). After transplantation, the alveolar-arterial oxygen gradient significantly decreased (from 17.3 +/- 7.1 mm Hg to 9 +/- 5.2 mm Hg; P < 0.001), as did the exhaled nitric oxide concentration (from 13 +/- 4.9 ppb to 6.2 +/- 2.8 ppb; P < 0.001). The decrease in the exhaled nitric oxide concentration was significantly correlated with the decrease in the alveolar-arterial oxygen gradient (r = 0.56; P = 0.014). Five patients met the criteria for the diagnosis of the hepatopulmonary syndrome before transplantation; the syndrome was cured by transplantation. CONCLUSIONS: The correlation between the decrease in exhaled nitric oxide concentration after liver transplantation and the improvement in oxygenation reinforces the hypothesis that nitric oxide is an important mediator of impaired oxygenation in patients with cirrhosis. PMID- 9735068 TI - The antiplatelet effects of ticlopidine and clopidogrel. AB - Ticlopidine and clopidogrel achieve antiplatelet effects by inhibiting the binding of adenosine 5'-disphosphate to its platelet receptor. Ticlopidine was first shown to decrease major events compared with placebo or aspirin in patients with stroke or recent transient ischemic attack. Randomized studies in patients undergoing coronary artery stenting have shown that ticlopidine reduces the risk for subacute stent thrombosis compared with warfarin-based regimens. Smaller studies have also shown this drug to have benefit during follow-up in patients with unstable angina, peripheral arterial disease, saphenous vein coronary bypass grafts, and diabetic retinopathy. Clopidogrel was recently approved by the U.S. Food and Drug Administration for the reduction of ischemic events in patients with recent myocardial infarction, stroke, or peripheral arterial disease (incidence, 5.32% per year compared with 5.83% per year for aspirin; P = 0.043) with no added risk for neutropenia. The combination of clopidogrel and aspirin, as well as the utility of clopidogrel in other patient populations and in stenting, requires further study. Ticlopidine and clopidogrel seem to have beneficial effects compared with aspirin (the current standard) in a broad range of patients. These observations highlight the importance of antiplatelet therapy in cardiovascular disease. PMID- 9735069 TI - Social conditions and self-management are more powerful determinants of health than access to care. AB - Professional organizations advocate universal access to medical care as a primary approach to improving health in the population. Access to medical services is critical to outcomes of acute processes managed in an inpatient hospital, the setting of most medical education, research, and training, but seems to be limited in its capacity to affect outcomes of outpatient care, the setting of most medical activities. Persistent and widening disparities in health according to socioeconomic status provide evidence of limitations of access to care. First, job classification, a measure of socioeconomic status, was a better predictor of cardiovascular death than cholesterol level, blood pressure, and smoking combined in employed London civil servants with universal access to the National Health Service. Second, disparities in health according to socioeconomic status widened between 1970 and 1980 in the United Kingdom despite universal access (similar trends were seen in the United States). Third, in the United States, noncompletion of high school is a greater risk factor than biological factors for development of many diseases, an association that is explained only in part by age, ethnicity, sex, or smoking status. Fourth, level of formal education predicted cardiovascular mortality better than random assignment to active drug or placebo over 3 years in a clinical trial that provides optimal access to care. Increased recognition of limitations of universal access by physicians and their professional societies may enhance efforts to improve the health of the population. PMID- 9735070 TI - Access to care is the centerpiece in the elimination of socioeconomic disparities in health. AB - Many health care professionals have sustained an almost single-minded conviction that disparities in access to health care across socioeconomic groups are the key reason for the major discrepancies in health status between wealthy persons and poor persons. Others, however, have argued that a host of factors work to create major impediments and that reducing or eliminating financial barriers to health care in particular will do little to reduce discrepancies in health status. This paper, while acknowledging the spectrum of contributing factors, argues that the elimination of financially based differences in access is central to any effort to create equity in outcomes across socioeconomic groups. Through selected review of the many studies on health insurance, access, outcomes, and socioeconomic status, it establishes that a core links affected populations, their difficulty in financing health care, and the threat to their well-being. In so doing, it cites findings that strongly associate lack of insurance (especially for persons who live in poverty), inability to obtain services, and adverse health outcomes. It also uses the example of Medicaid and other coverage for HIV-infected persons in particular as an important positive instance in which leveling the discrepancies in health care across socioeconomic groups can move toward creating quality in access and outcomes. The competitive pressures in today's health care environment threaten to drive socioeconomic groups further apart, especially insured and uninsured persons. However, the recent enactment of state actions, especially the State Child Health Insurance Program, represent powerful examples of health insurance expansion that have lessons for policymakers at all levels for the monitoring and reduction of socioeconomic disparities. PMID- 9735071 TI - The use and misuse of classification and diagnostic criteria for complex diseases. PMID- 9735072 TI - Medical care and health improvement: the critical link. PMID- 9735073 TI - Guidelines for the clinical diagnosis of Lyme disease. PMID- 9735074 TI - Guidelines for the clinical diagnosis of Lyme disease. PMID- 9735075 TI - Guidelines for the clinical diagnosis of Lyme disease. PMID- 9735076 TI - Jugular vein sampling in Cushing disease. PMID- 9735077 TI - Treating familial Mediterranean fever with prazosin hydrochloride. PMID- 9735078 TI - Increased synovial interleukin-8 and interleukin-6 levels in pseudogout associated with granulocyte colony-stimulating factor. PMID- 9735079 TI - The medical education of William Brooks Bigler (1863). PMID- 9735080 TI - Incidence of and risk factors for ventilator-associated pneumonia in critically ill patients. AB - BACKGROUND: Understanding the risk factors for ventilator-associated pneumonia can help to assess prognosis and devise and test preventive strategies. OBJECTIVE: To examine the baseline and time-dependent risk factors for ventilator associated pneumonia and to determine the conditional probability and cumulative risk over the duration of stay in the intensive care unit. DESIGN: Prospective cohort study. SETTING: 16 intensive care units in Canada. PATIENTS: 1014 mechanically ventilated patients. MEASUREMENTS: Demographic and time-dependent variables reflecting illness severity, ventilation, nutrition, and drug exposure. Pneumonia was classified by using five methods: adjudication committee, bedside clinician's diagnosis, Centers for Disease Control and Prevention definition, Clinical Pulmonary Infection score, and positive culture from bronchoalveolar lavage or protected specimen brush. RESULTS: 177 of 1014 patients (17.5%) developed ventilator-associated pneumonia 9.0 +/- 5.9 days (median, 7 days [interquartile range, 5 to 10 days]) after admission to the intensive care unit. Although the cumulative risk increased over time, the daily hazard rate decreased after day 5 (3.3% at day 5, 2.3% at day 10, and 1.3% at day 15). Independent predictors of ventilator-associated pneumonia in multivariable analysis were a primary admitting diagnosis of burns (risk ratio, 5.09 [95% CI, 1.52 to 17.03]), trauma (risk ratio, 5.00 [CI, 1.91 to 13.11]), central nervous system disease (risk ratio, 3.40 [CI, 1.31 to 8.81]), respiratory disease (risk ratio, 2.79 [CI, 1.04 to 7.51]), cardiac disease (risk ratio, 2.72 [CI, 1.05 to 7.01]), mechanical ventilation in the previous 24 hours (risk ratio, 2.28 [CI, 1.11 to 4.68]), witnessed aspiration (risk ratio, 3.25 [CI, 1.62 to 6.50]), and paralytic agents (risk ratio, 1.57 [CI, 1.03 to 2.39]). Exposure to antibiotics conferred protection (risk ratio, 0.37 [CI, 0.27 to 0.51]). Independent risk factors were the same regardless of the pneumonia definition used. CONCLUSIONS: The daily risk for pneumonia decreases with increasing duration of stay in the intensive care unit. Witnessed aspiration and exposure to paralytic agents are potentially modifiable independent risk factors. Exposure to antibiotics was associated with low rates of early ventilator-associated pneumonia, but this effect attenuates over time. PMID- 9735081 TI - Opening the black box: how do physicians communicate about advance directives? AB - BACKGROUND: The quality of communication that leads to the completion of written advance directives may influence the usefulness of these documents, but the nature of that communication remains relatively unexplored. OBJECTIVE: To describe how physicians discuss advance directives with patients. DESIGN: Prospective study. SETTING: Five outpatient primary care medicine practices in Durham, North Carolina, and Pittsburgh, Pennsylvania. PARTICIPANTS: 56 attending internists and 56 of their established patients. Eligible patients were at least 65 years of age or had a serious medical illness. MEASUREMENTS: Two raters coded transcripts of audiotaped discussions about advance directives to document how physicians introduced the topic of advance directives, discussed scenarios and treatments, provided information, elicited patient values, and identified surrogate decision makers. RESULTS: Conversations about advance directives averaged 5.6 minutes; physicians spoke for two thirds of this time. In 91% of cases, physicians discussed dire scenarios in which most patients would not want to be treated, and 48% asked patients about their preferences in reversible scenarios. Fifty-five percent of physicians discussed scenarios involving uncertainty, typically using vague language. Patients' values were rarely explored in detail. In 88% of cases, physicians discussed surrogate decision making and documents to aid in advance care planning. CONCLUSIONS: Although they accomplished the goal of introducing patients to advance directives, discussions infrequently dealt with patients' values and attitudes toward uncertainty. Physicians may not have addressed the topic in a way that would be of substantial use in future decision making, and these discussions did not meet the standards proposed in the literature. PMID- 9735082 TI - Cumulative epinephrine dose during cardiopulmonary resuscitation and neurologic outcome. AB - BACKGROUND: Epinephrine is the drug of choice in advanced cardiac life support, but it can have deleterious side effects after restoration of spontaneous circulation. OBJECTIVE: To investigate the association between the cumulative epinephrine dose used in advanced cardiac life support and neurologic outcome after cardiac arrest. DESIGN: Retrospective cohort study. SETTING: University hospital. PATIENTS: Adults admitted to the emergency department with witnessed, nontraumatic, normothermic ventricular fibrillation cardiac arrest and unsuccessful initial defibrillation. MEASUREMENTS: Functional neurologic outcome was regularly assessed by cerebral performance category (CPC) within 6 months after cardiac arrest. A CPC of 1 or 2 was defined as favorable recovery. RESULTS: Among 178 enrolled patients, the median cumulative epinephrine dose administered was 4 mg (range, 0 to 50 mg). In 151 patients (84%), spontaneous circulation was restored; 63 of these 151 patients (42%) had favorable neurologic recovery. Patients with an unfavorable CPC received a significantly higher cumulative dose of epinephrine than did patients with a favorable CPC (4 mg compared with 1 mg; P < 0.001). This finding persisted after stratification by duration of resuscitation. After possible cofounders were controlled for, the cumulative epinephrine dose remained an independent predictor of unfavorable neurologic outcome. CONCLUSIONS: The results indicate that an increasing cumulative dose of epinephrine administered during resuscitation is independently associated with unfavorable neurologic outcome after ventricular fibrillation cardiac arrest. PMID- 9735083 TI - Increased serum lipoprotein(a) levels in patients with early renal failure. AB - BACKGROUND: Elevated serum lipoprotein(a) levels have been found in patients with end-stage renal disease and in patients undergoing dialysis, suggesting that this lipoprotein contributes to the increased cardiovascular risk seen in these patients. It is not known whether lipoprotein(a) levels are elevated in the early phases of renal disease. OBJECTIVE: To evaluate levels of lipoprotein(a) and other lipids and the prevalence of atherosclerotic disease in patients with early renal failure. DESIGN: Cross-sectional study. SETTING: Hypertension clinic of a university medical center. PATIENTS: 257 patients with normal renal function and 160 patients with early impairment of renal function (creatinine clearance, 30 to 89 mL/min per 1.73 m2 of body surface area). MEASUREMENTS: Renal function was assessed by 24-hour creatinine clearance, proteinuria, and microalbuminuria. Cardiovascular disease status was also assessed. Serum lipoprotein(a), lipids, apolipoproteins, and apolipoprotein(a) isoforms were measured. RESULTS: Age, blood pressure, and serum lipoprotein(a) levels were greater in patients with early renal failure than in those with normal renal function and were independently associated with the presence of decreased creatinine clearance. Serum lipoprotein(a) and creatinine clearance were inversely correlated. The prevalence of coronary artery, cerebrovascular, and peripheral vascular disease was greater in patients with early renal failure than in those with normal renal function. The frequency distribution of apolipoprotein(a) isoforms was similar in patients with normal and those with impaired renal function. CONCLUSIONS: Serum lipoprotein(a) levels are elevated in patients with early impairment of renal function and are associated with greater prevalence of cardiovascular disease. An inverse correlation between serum lipoprotein(a) level and creatinine clearance and a frequency distribution of apolipoprotein(a) isoforms similar to that of normal patients point to decreased renal catabolism as a probable mechanism of lipoprotein(a) elevation in patients with early renal failure. PMID- 9735084 TI - Reinfection with the agent of human granulocytic ehrlichiosis. PMID- 9735085 TI - Update in infectious diseases. PMID- 9735086 TI - Management of pituitary tumors. AB - Management of pituitary tumors has improved in the past decade since the introduction of novel therapeutic agents. As a result, several treatment options are now available. Dopamine agonists are the preferred treatment for both symptomatic microprolactinomas and macroprolactinomas; these drugs result in normalization of hormone levels and tumor shrinkage in most treated patients. New formulations (such as cabergoline and parenteral bromocriptine) with prolonged duration of action offer improved compliance with treatment and cure rates. For acromegaly and adrenocorticotropin hormone (ACTH)-secreting, thyroid-stimulating hormone (TSH)-secreting, and nonfunctional adenomas, surgery often results in cure. Octreotide and the long-acting, slow-release somatostatin analogues are effective medical alternatives to or adjuvants for transsphenoidal surgery in patients with growth hormone-secreting and TSH-secreting tumors. No drug treatment is available for symptomatic nonfunctional tumors, and patients with ACTH-secreting adenomas may benefit from cortisol-lowering drugs after surgical failure. Pituitary irradiation may be required after surgery for ACTH-secreting, TSH-secreting, and nonfunctioning tumors; it is less commonly required for acromegaly. Although many pituitary tumors are successfully resected, functional adenomas may not be cured by surgery. As more-effective drugs are introduced for the management of pituitary tumors, more patients with hormone-secreting adenomas are being successfully treated medically. PMID- 9735087 TI - Multiple endocrine neoplasia type 1: clinical and genetic topics. AB - Multiple endocrine neoplasia type 1 (MEN1) consists of benign, and sometimes malignant, tumors (often multiple in a tissue) of the parathyroids, enteropancreatic neuroendocrine system, anterior pituitary, and other tissues. Skin angiofibromas and skin collagenomas are common. Typically, MEN1 tumors begin two decades earlier than sporadic tumors. Because of tumor multiplicity and the tendency for postoperative tumor recurrence, specialized methods have been developed for preoperative and intraoperative localization of many MEN1 associated tumors. The MEN1 gene was recently isolated by positional cloning. This strategy progressively narrows the size of the candidate MEN1 gene interval on the chromosome and then finds and tests many or, if needed, all genes within that interval. The MEN1 gene was finally identified because it was the one gene that contained mutations in most DNAs from a test panel of MEN1 cases. It has been suggested that MEN1, like many hereditary cancer syndromes, is caused by mutation in a tumor suppressor gene that contributes to neoplasia when both gene copies in a tumor precursor cell have been sequentially inactivated ("two-hit" oncogenesis mechanism). Germline MEN1 mutations were found in most families with MEN1 and in most cases of sporadic MEN1. In addition, the MEN1 gene was the gene most likely to show acquired mutation in several sporadic or nonhereditary tumors parathyroid adenomas, gastrinomas, insulinomas, and bronchial carcinoids. Most germline or acquired MEN1 mutations predicted truncation (and thus likely inactivation) of the encoded protein, supporting expectations for the "first hit" to a tumor suppressor gene. Testing for MEN1 germline mutation is possible in a research setting. Candidates for MEN1 mutation testing include patients with MEN1 or its phenocopies and first-degree relatives of persons with MEN1. PMID- 9735088 TI - Electronic patient-physician communication: problems and promise. AB - A critical mass of Internet users will soon enable wide diffusion of electronic communication within medical practice. E-mail between physicians and patients offers important opportunities for better communication. Linking patients and physicians through e-mail may increase the involvement of patients in supervising and documenting their own health care, processes that may activate patients and contribute to improved health. These new linkages may have profound implications for the patient-physician relationship. Although the federal government proposes regulation of telemedicine technologies and medical software, communications technologies are evolving under less scrutiny. Unless these technologies are implemented with substantial forethought, they may disturb delicate balances in the patient-physician relationship, widen social disparities in health outcomes, and create barriers to access to health care. This paper seeks to identify the promise and pitfalls of electronic patient-physician communication before such technology becomes widely distributed. A research agenda is proposed that would provide data that are useful for careful shaping of the communications infrastructure. The paper addresses the need to 1) define appropriate use of the various modes of patient-physician communication, 2) ensure the security and confidentiality of patient information, 3) create user interfaces that guide patients in effective use of the technology, 4) proactively assess medicolegal liability, and 5) ensure access to the technology by a multicultural, multilingual population with varying degrees of literacy. PMID- 9735089 TI - The next chapter in the high-dose epinephrine story: unfavorable neurologic outcomes? PMID- 9735090 TI - The era of adherence to HIV therapy. PMID- 9735091 TI - Hepatitis A: a potentially serious disease. PMID- 9735092 TI - Hepatitis A: a potentially serious disease. PMID- 9735093 TI - Hepatitis A: a potentially serious disease. PMID- 9735094 TI - Vancomycin-associated linear IgA bullous dermatosis. PMID- 9735095 TI - Linear IgA bullous dermatosis associated with vancomycin. PMID- 9735096 TI - Regression of metastatic carcinoma of the skin appendages after intralesional granulocyte-macrophage colony-stimulating factor. PMID- 9735097 TI - When doctors get sick. PMID- 9735098 TI - Empirical therapy for community-acquired pneumonia. PMID- 9735099 TI - Number of uninsured continues to rise. PMID- 9735100 TI - Remembering medicine's past. PMID- 9735101 TI - [Feed intake and milk production in dairy cows fed whole fat soybeans]. AB - In two experiments (E 1, E 2) two different basic diets were fed to 36 diary cows with addition of 1 or 2 kg of full fat soya beans (SB) or without SB for six weeks. The cows were fed individually. The basic diet consisted of 49% grass silage, 23% maize silage and 21% hay, supplemented with 7% manioc (% DM) in E 1 and of grass, supplemented with 2.0 kg maize silage, 3.0 kg hay and 1.5 kg manioc (kg DM/cow.d) in E 2. The basic diet used in E 1 and the grass in E 2 were offered ad libitum. Further a mixture of 1 kg SB, soya bean meal and manioc (Treatment 1), a mixture of 2 kg SB and manioc (Treatment 2) and a mixture of soya bean meal and manioc (Treatment 3, control), respectively, was supplemented to achieve diets with equivalent concentrations of energy and protein. Depending on the daily milk yield a concentrate was fed to high yielding cows (> 21.5 kg milk in E 1 and > 25.0 kg milk in E 2, respectively). The addition of 1 kg SB increased the forage intake in both experiments by 1.2-1.5 kg DM/cow.d significantly, while the addition of 2 kg SB increased the forage intake only in E1 to the same extent. The mean total DMI was 19.2 kg in E 1 and 17.7 kg in E 2, respectively. The daily milk yield was not influenced by addition of SB. The average milk yield was 28.8 kg and 26.7 kg in E 1 and E 2, respectively. The milk fat content increased significantly in E 1 by 0.3-0.4% with no respect to the amount of the supplied SB (4.11% vs. 3.74%). In E 2 the milk fat content increased only slightly (3.84% vs. 3.74%). Milk protein content was not influenced in E 1, but decreased in E 2 after addition of 2 kg SB. PMID- 9735102 TI - [Systemic availability of bovine immunoglobulin G and chicken immunoglobulin Y after feeding colostrum and whole egg powder to newborn calves]. AB - In connection with a study on the prophylaxis of infectious diarrhea with specific egg yolk antibodies, the systemic availability of colostral bovine immunoglobulin G (bIgG) and chicken immunoglobulin Y (IgY) after feeding egg powder was investigated on 26 newborn calves from 23 different farms. Blood was sampled daily and at the same day time from these calves in the first 14 days of life. During the feeding of colostrum, the mean bIgG concentration was highest at day 1 post natum with a value of 9.3 mg/ml serum. Thereafter, the mean bIgG level was reduced continuously to a significant lower concentration of 4.9 mg/ml serum at day 12 post natum and remained nearly constant at 5.2 mg/ml till to the end of the observation period. Total protein concentrations in the serum did not change and plateaued at a mean value of 56.2 mg/ml (SD 11.2). The number of colostrum meals had no significant effect on the mean bIgG concentrations during that period. The individual variation of bIgG concentrations was very high on every day of the sampling period. The mean coefficient of variation was at 52.1 % (SD 5.7). After having described the individual bIgG concentration curves mathematically with a regression curve, two groups with significantly different bIgG elimination constants (k) could be obtained. Thus in one group (n = 10) with k-values of < -0.02 a mean half time of serum bIgG of 24.3 days (SD 4.6) was calculated. In the other group of calves (n = 16) with elimination constants of k > -0.02, a mean half time of 68.5 days (SD 36.7) could be calculated, possibly because these calves started earlier with their endogenous bIgG production. Additionally, to 18 of these calves 20 g egg powder with an IgY concentration of 15 mg/g was fed up to day 14. Calves had a maximal mean IgY concentration of 1.9 micrograms/ml serum if egg powder feeding started already during the first 12 hours of life. Starting at a later time resulted in a significant reduction of IgY levels. For example, the mean initial IgY concentration dropped to 0.035 micrograms/ml serum after having had the first egg powder application between 25 and 48 hours post natum. Using the individual IgY elimination constant derived from a regression analysis (r2 = 0.84) of the IgY concentration curve, a mean IgY half time of 5.0 days (SD 2.5) could be calculated. To prevent the absorption of heterologous antibodies and consecutively, also to prevent a possible systemic effect, egg powder for prophylactic purposes in newborn calves should be fed after the first 24, better 48 hour, post natum. Most important for the prophylactic effect of specific antibodies on infectious diarrhea is not their systemic but their high local intestinal availability. PMID- 9735103 TI - Kinetic analysis of an inhibitor-resistant variant of the OHIO-1 beta-lactamase, an SHV-family class A enzyme. AB - The Met69-->Ile mutant of the OHIO-1 beta-lactamase, an SHV-family enzyme, is resistant to inactivation by beta-lactamase inhibitors. Analysis of purified Met69-->Ile enzyme reveals that its isoelectric point (pI 7.0) and CD spectrum are identical with those of the OHIO-1 enzyme. Levels of beta-lactamase expression in Escherichia coli as determined by immunoblotting are similar for OHIO-1 and Met69-->Ile beta-lactamase. The kinetic constants of the Met69-->Ile enzyme compared with OHIO-1 are smaller for benzylpenicillin (Km = 6 microM compared with 17 microM; kcat = 234 s-1 compared with 345 s-1 respectively) and carbenicillin (Km = 3 microM compared with 17 microM; kcat = 131 s-1 compared with 320 s-1 respectively). For the cephalosporins cephaloridine and 7-(thienyl- 2-acetamido)-3-[2-(4-N,N- dimethylaminophenylazo)pyridinium-methyl]-3-cephem-4 carboxylic acid (PADAC), a similar pattern is also seen (Km=38 microM compared with 96 microM and 6 microM compared with 75 microM respectively; kcat = 235 s-1 compared with 1023 s-1 and 9 s-1 compared with 50 s-1 respectively). Consistent with minimum inhibitory concentrations that show resistance to beta-lactam beta lactamase inhibitors, the apparent Ki values, turnover numbers and partition ratios (kcat/kinact) for the mechanism-based inactivators clavulanate, sulbactam and tazobactam are increased. The inactivation rate constants (kinact) are decreased. The difference in activation energy, a measurement of altered affinity for the wild-type and mutant enzymes leading to acylation of the active site, reveals small energy differences of less than 8.4 kJ/mol. In total, these results suggest that the Met-->Ile substitution at position 69 in the OHIO-1 beta lactamase alters the active site, primarily affecting the interactions with beta lactamase inhibitors. PMID- 9735104 TI - Signalling molecules and the regulation of intracellular transport. AB - A variety of signalling molecules has been implicated over the past 8 years in the regulation of intracellular transport pathways. Those molecules include heterotrimeric GTP binding proteins, members of the protein kinase C family, and members of the Rho subfamily of small GTPases. Until recently, no common theme among the three classes of regulators was apparent. The finding that all three can influence the activity of phospholipase D (PLD), and the fact that members of the Arf subfamily of GTPases (with established roles in intracellular transport) are potent activators of PLD suggests the hypothesis that PLD is a focal point for integration of cellular responses to hormone signalling and for membrane homeostasis. Work during the past 2 years is beginning to uncover some transport pathways where PLD involvement is inferred. It is proposed that, if signalling is required to monitor and adjust transport rates to and from the various membrane organelles, the most economical way to achieve this would be to regulate recycling and allow the concentration of cargo receptors to determine forward transport. PMID- 9735105 TI - Histologic evaluation of neck dissection specimens. AB - Histologic evaluations of neck dissection specimens from carcinomas of the head and neck provide information required for disease staging, planning further treatment, and prognosis. Histologic evaluation performed adequately and accurately can and continues to provide a more accurate, meaningful, and promising means of formulating and predicting prognosis including risk of metastases. A multi-institutional study using comprehensive standardized histologic evaluation of histopathologic variables of primary tumor and cervical lymph nodes among homogenous patient groups receiving similar therapy is important. Histopathologic parameters remain an important adjunct parameter to clinical evaluation in guiding, planning treatment, and predicting prognosis for patients with head and neck cancers. PMID- 9735106 TI - Management of the neck in patients with head and neck cancer treated by concurrent chemotherapy and radiation. AB - The current high level of interest in organ preservation strategies for patients with advanced squamous cell carcinoma of the head and neck undoubtedly will result in increasing numbers of patients managed initially with chemotherapy and radiation, either sequentially or concurrently. In some protocols, surgery, and neck dissection in particular, will either be mandatory or offered based on the degree of response to treatment and initial stage of neck disease. Head and neck oncologic surgeons need to be involved and at the forefront of such trials, to allow meaningful data regarding pathologic response to treatment to be obtained, as well as to define the role of surgery in such patients. Although present data is limited, it would appear that in patients achieving a complete response to chemoradiation, the role of neck dissection may be more limited than in the past, even for patients with N2 to N3 neck disease at presentation. Surgical complications may be increased in this heavily treated patient population, and subsequent surgery should be designed to minimize the risk of wound complications, especially if performed before the patient has made a full recovery from the metabolic and immunologic derangements associated with chemoradiation. Head and neck surgeons need to play an active role in the design and conduct of chemoradiation trials so that these and other relevant questions will be answered by the data generated. PMID- 9735107 TI - Management of the neck in nasopharyngeal carcinoma (NPC). AB - Management of cervical nodal metastasis from nasopharyngeal carcinoma (NPC) begins with a thorough assessment of the patient to determine extent of the disease process at the primary site, regionally and systematically. Detailed knowledge of the anatomy of the head and neck will facilitate an accurate diagnosis and subsequent staging of each individual patient. The use of the appropriate diagnostic tools such as imaging, fine-needle aspiration studies, and serology direct the clinician to the appropriate management scheme. This article attempts to cull information from various clinicians who treat the majority of NPC patients, and to raise the issue of the need for more innovative approaches. PMID- 9735108 TI - Elective irradiation of the N0 neck in squamous cell carcinoma of the upper aerodigestive tract. AB - The decision of how to optimally manage the clinically negative neck is based on the likelihood of clinically inapparent disease and the efficacy of salvage therapy. The criteria of decision for elective management of the neck takes into account the site, size, depth of infiltration, grading of the primary lesion, clinical and radiologic evaluation, and patient wishes. Diagnostic procedures currently used in evaluating head and neck cancer patients with nodal disease are reviewed. Elective irradiation of the N0 neck in patients with squamous cell carcinoma of the head and neck is an effective means of maintaining locoregional control. The impact of elective nodal treatment on disease free survival and overall survival is discussed. PMID- 9735109 TI - Neck dissection in the treatment of cancer of major salivary glands. AB - The treatment of the neck nodes in salivary gland tumors has changed considerably in the last two decades. The current thinking and the rationale for it are discussed in detail in this article. PMID- 9735111 TI - Treatment of the neck in melanoma. AB - The treatment of the neck/parotid in melanoma of the head and neck has changed in recent years. With the use of ultrasound, fine-needle aspiration, lymphoscintigraphy, modified/selective dissection, adjuvant radiation therapy, and systemic interferon, patients with suspected metastatic melanoma in the neck have a better opportunity to have early diagnosis, preservation of function, and improved regional control and survival. PMID- 9735110 TI - Management of the neck in thyroid cancer. AB - The incidence of nodal metastasis in differentiated thyroid cancer ranges between 40% to 75%. Elective neck dissection is generally not advised in patients with differentiated thyroid cancer; however, if clinically apparent nodal disease is noted in the tracheoesophageal groove during surgery, central compartment clearance is advised. If clinically apparent nodal disease is present in the lateral compartment of the neck, modified neck dissection preserving the sternomastoid, accessory nerve, and jugular vein is advised. The "berry picking procedure" is generally not recommended because of the higher incidence of regional recurrence. Due consideration should be given for parathyroidal transplantation if the blood supply to the parathyroids is damaged during central compartment clearance. The incidence of lymph node metastasis is highest in young patients, however, lymph node metastasis has no bearing on long-term survival. There seems to be a higher incidence of regional recurrence in elderly individuals. If patients present with bulky nodal disease, consideration may be given for postoperative radioactive iodine dosimetry and ablation if necessary. Differentiated thyroid cancer represents a unique disease in the human body, where lymph node metastasis has no prognostic implication. Aggressive surgical clearance is advised in patients with medullary thyroid cancer in the central compartment and the jugular chain lymph nodes. PMID- 9735112 TI - Management of the neck in nonmelanocytic cutaneous carcinomas. AB - Cutaneous malignancies are the most common causes of cancer in the United States, and the preponderance occur in the head and neck region. From 0.3% to 13.7% of cutaneous squamous cell carcinomas and from 0.0028% to 0.4% of cutaneous basal cell carcinomas metastasize to the cervical nodes. A description of the head and neck cutaneous lymphatic drainage is presented, followed by recommendations regarding neck dissection modifications appropriate to the primary sites and nodal eschalons involved. PMID- 9735113 TI - Advances and new concepts in oral and maxillofacial pathology AB - New techniques in surgical pathology at the cellular and molecular levels offer the clinician help in determining modalities of treatment of specific diseases. In addition to routine staining, adjunctive tests such as immunohistochemical analysis, and the various methods of evaluating nucleic acid have helped make this possible. The efficacy of fine-needle aspiration biopsy has been enhanced by these diagnostic aids that enable the assessment of information from small amounts of tissue. PMID- 9735114 TI - Medical oncology in the management of head and neck cancers AB - General approaches and therapeutic goals of medical oncology for head and neck cancer are presented. The effectiveness of chemotherapy for the treatment of different stages of head and neck cancer in specific anatomic sites is discussed, as well as complications associated with chemotherapy, and approaches to the prevention and management of these stages. Systemic side effects that may occur with specific therapeutic agents are presented in a tabular format. Future directions and evolving approaches to head and neck cancer therapy are summarized. PMID- 9735115 TI - A brief review of the immune system. AB - This article provides a brief review of the immune system and describes the features of innate and adaptive immunity and their similarities and differences. The mechanism of antigen presentation and major histocompatibility complex restriction is discussed as well as the structure and function of T cells and B cells. Three tables present a concise description of cytokines, interleukins, and chemokines. PMID- 9735116 TI - Primary immunodeficiency disorders. AB - The primary immunodeficiency diseases are a relatively rare group of congenital disorders that are linked by the expression of an excessive number, duration, or severity of infections. The clinical features of most of the primary immunodeficiency diseases have been well described by astute physicians over several decades and have provided important clues to our basic understanding of human immunology. In contrast, the genetic basis and potential life-saving therapies for many of these disorders have been established only over the past few years. These recent advances have resulted in the prognosis of many of these disorders being largely dependent on their rapid recognition and treatment. Increased awareness of the differentiating epidemiologic, clinical, laboratory and genetic features of these diseases hold the promise of both furthering our understanding of basic human immunology and providing improved care for this challenging group of patients. PMID- 9735117 TI - HIV infection and AIDS. AB - The entry of one HIV virion into a human being has the potential to cause death by the inexorable replication of the virus within the principal T lymphocyte, the CD4+ T cell. Although combination antiretroviral therapy, particularly therapy with protease inhibitors, decreases the viral burden to very low, even undetectable, levels, sequestration of the virus in privileged sites, including a long-lived CD4+ T cell, has frustrated efforts at eradication of HIV. Activation of the immune system, therefore, appears essential before this infection can be conquered. Powerful vaccines capable of preventing infection remain the hope of the world. PMID- 9735118 TI - Allergic skin disease: atopic dermatitis as a prototype. AB - The skin is the largest immune organ in the body. It is strategically positioned as an interface between a hostile antigenic world and a complex immune system characterized by inflammatory cells, mediators, lymphocytes, and a panoply of cytokines. These cytokines act as subcellar messengers that direct effector lymphocytes, eosinophils, mast cells, and a host of other cells to target the skin in allergic inflammation. Just as asthmas and allergic rhinitis are prototypes of IgE-mediated respiratory disease, atopic dermatitis is the prototype allergic skin disease. This article focuses on an understanding of the pathogenic mechanisms involved in atopic dermatitis and outlines the clinical spectrum of cutaneous allergic disease. Finally, a state-of-the-art approach to treatment is offered to the clinician confronted with the management of this difficult disorder. PMID- 9735119 TI - Drug allergies. AB - Allergic drug reactions are a significant cause of morbidity and mortality. Because it is difficult to identify the culprit drug and the underlying pathophysiologic mechanisms involved in these reactions, a systematic approach should be adopted in the evaluation of drug-allergic patients. Initially, the type of reaction should be determined. It should be realized that not all adverse reactions are allergic in nature. Allergic drug reactions comprise only a small category of adverse reactions in general. Therefore, the physician must determine if the reaction demonstrates features common to immunologic reaction. Subsequently, a detailed history should be obtained and a physical should be performed. Important information includes medication usage, previous drug exposure, current illness, family history of drug allergy and personal history of drug allergy. In managing the drug-allergic patient, the physician may choose to: select an alternative, non-cross-reacting drug if future therapy is needed; premedicate prior to future drug exposure if such regimens have been shown to be effective; or consult an allergist regarding the potential graded challenge or desensitization. PMID- 9735120 TI - Anaphylaxis: diagnosis and treatment. AB - Anaphylaxis is a clinical syndrome and is caused by many different agents. Acute therapy is epinephrine. Chronic therapy depends on the agent causing the anaphylaxis, but usually consists of avoidance of the agent only. Individuals who suffer from this problem should be issued epinephrine and wear or carry medical alert bracelets. PMID- 9735121 TI - Food allergy. AB - Food allergies are immunologic reactions to food allergens or food components. Several distinct clinical entities fall under this term, including immediate-in time allergic reactions, which are IgE-dependent and involve mast cells and basophils, and delayed-in-time reactions to foods, such as food-induced enterocolitis, which involve additional effector systems. Most food allergies are precipitated by a small number of foods. The diagnosis of these diseases depends on history, physical examination, specific diagnostic assays, and oral food challenge. The differential diagnosis of these diseases is extensive. Treatment of food allergies relies on identification of the food substance that induces the reaction and subsequent avoidance measures. When an individual inadvertently consumes food to which he or she is sensitized, pharmacologic treatment is available. PMID- 9735122 TI - Diagnosis and management of rhinitis. AB - This article reviews the differential diagnoses for rhinitis, medications available for the treatment of rhinitis, and special circumstances (such as pregnancy or medication side-effects) that may influence a clinician's decision. Considering the economic impact of rhinitis, the cost of prescription medications, and quality-of-life issues that are affected by rhinitis, physicians dealing with managed care organizations should make their diagnosis and treatment decisions carefully. PMID- 9735123 TI - Bronchial asthma: update on the revised guidelines for diagnosis and management. AB - Bronchial asthma is a chronic disease with variable airway narrowing, respiratory distress, hyper-responsiveness and inflammation. The morbidity and mortality are increasing despite availability of newer diagnostic and therapeutic strategies. The National Institutes of Health recently issued revised guidelines for disease management. The keys to improved care include earlier recognition of the illness, reduced exposure to triggers, careful monitoring, greater use of long-term control medications and improved patient education. PMID- 9735124 TI - Allergen immunotherapy and avoidance. AB - Since its introduction almost a century ago, immunotherapy continues to be an effective method of managing allergic rhinitis, allergic asthma, and insect anaphylaxis. Confusion and misinformation on the part of physicians and the public lead to the inappropriate use of this treatment. Before immunotherapy is started, appropriate avoidance techniques and pharmacotherapy should be instituted (except for insect anaphylaxis, where immunotherapy is a part of the initial treatment.) When these measures fail, or significant side effects are encountered, immunotherapy can be beneficial. This article eliminates as much as possible the ambiguity surrounding immunotherapy to help the clinician understand more clearly the appropriate use of immunotherapy and the treatment of allergic diseases. PMID- 9735125 TI - Future role of the allergist-immunologist. AB - The future role of the allergist/immunologist is dictated by current training and research, which promise to broaden the expertise of practitioners of this discipline. Specialists in allergy and immunology are certified by the American Board of Allergy and Immunology following specialized training in allergic diseases, clinical immunology, procedural skills, clinical principles and analytic methods, clinical laboratory test proficiency, and research. The expanding role of clinical immunology in the management of multiple diseases suggests that specialists in allergy and immunology will continue to broaden their expertise in the care of disorders with an immune component. The discipline of allergy and immunology offers a number of career advantages in both the practice of allergy and immunology and in the application of training in allergy and immunology to opportunities in the pharmaceutical industry and academics. Manpower assessments suggest that the number of allergists and immunologists in training are insufficient to meet the current needs. PMID- 9735126 TI - Papillary carcinoma of the thyroid gland: treatment based on risk group definition. AB - This article outlines the current controversy regarding the extent of treatment of papillary carcinoma of the thyroid. It emphasizes that following general principles in surgical oncology, surgeons doing endocrine cancer management should scale the extensiveness of their operative procedure to the risk of the cancer. The majority of patients with papillary carcinoma fall into a low-risk group with a 1% or less risk of death over a period of many years. It is illogical to treat these patients in the same fashion as patients who have a 50% risk of death in a shorter period of time. The various components of the risk assignment systems are outlined and emphasized to give surgeons the confidence to use risk category in their selection of surgical and adjuvant treatment of papillary carcinoma of the thyroid. PMID- 9735127 TI - Papillary thyroid carcinoma: justification for total thyroidectomy and management of lymph node metastases. AB - Papillary thyroid carcinoma (PTC) is the most common epithelial thyroid tumor and comprises approximately 80% of all thyroid cancers. In this article, the authors discuss the data showing that total thyroidectomy is the treatment of choice of clinically significant PTC, and review an algorithm for the management of lymph node metastases. Although the prognosis for patients with PTC is generally good, appropriate surgical management (total thyroidectomy plus 131I and life-long TSH suppression) can further reduce recurrence and cancer death rates significantly. PMID- 9735128 TI - Guidelines for the use of radio-iodine, thyroid hormone, and treatment of metastatic disease in patients with differentiated thyroid cancer. AB - The treatment of metastatic differentiated thyroid carcinoma is currently at a crossroad. The stunning effect of imaging doses of RAI on subsequent treatment doses is being recognized. Alternatives to RAI imaging for diagnostic purposes are being tested; these include ultrasonography for local and cervical nodal remnants/recurrences, Sestamibi and other isotope scanning that do not require the discontinuation of TSH suppression, and the measurement of circulating thyroglobulin that is rapidly becoming the cornerstone of the detection and the treatment follow-up of metastatic carcinoma. PMID- 9735129 TI - Medullary thyroid carcinoma: genetic advances, treatment recommendations, and the approach to the patient with persistent hypercalcitoninemia. AB - Medullary thyroid cancer is a tumor of the thyroid C cells that occurs in sporadic and hereditary clinical settings. Genetic testing of at-risk individuals is available and has been applied to patient management. Plasma calcitonin levels are a sensitive marker for the presence of disease. Surgery offers the best hope for cure and also is an effective modality for managing metastatic and recurrent disease. PMID- 9735130 TI - Anaplastic cancer, lymphoma, and metastases of the thyroid gland. AB - Not all thyroid carcinomas behave in an indolent fashion. There is a rare group of thyroid neoplasms that is biologically aggressive and carries a poor prognosis. These tumors include poorly differentiated tumors, anaplastic, and thyroid lymphomas. Poorly differentiated tumors are important tumors to recognize because aggressive surgical intervention appears to offer the best chance for long-term survival. Until recently, anaplastic carcinoma was universally fatal. Preoperative chemotherapy and hyperfractionated radiation has led to better local control and a few long-term survivors. The role of the surgeon in the treatment of the thyroid lymphomas remains controversial, however, most would agree that the surgeons role is limited, because these tumors are radiosensitive and chemosensitive. PMID- 9735131 TI - Parathyroid adenoma, hyperplasia, and carcinoma: localization, technical details of primary neck exploration, and treatment of hypercalcemic crisis. AB - The pathologic characteristics and clinical presentation of patients with primary hyperparathyroidism are discussed including the treatment of hypercalcemic crisis. Surgical issues, including the use of localizing studies, and the surgical treatment of primary hyperparathyroidism are reviewed. PMID- 9735132 TI - The adrenal incidentaloma: guidelines for evaluation and recommendations for management. AB - Adrenal masses are identified incidentally on up to 1.5% of all abdominal CT scans. The appropriate evaluation and management of these "incidentalomas" remains controversial and centers on questions of function and potential for malignancy. Functional evaluation includes consideration of the diagnoses of aldosteronoma, pheochromocytoma, and corticosteroid-producing adenoma. Potential for malignancy can be evaluated using a number of imaging modalities, although none is diagnostic. Size remains one of the best criteria for assessing potential for malignancy. PMID- 9735133 TI - Benign and malignant pheochromocytoma: diagnosis, treatment, and follow-Up. AB - The clinical manifestation of pheochromocytoma is presented with specific emphasis on accurate diagnostic approach. The sensitivity, specificity, and indication of localizing studies of pheochromocytomas are reviewed. Management of benign and malignant pheochromocytomas are discussed with follow-up recommendations. PMID- 9735134 TI - Functioning and nonfunctioning adrenocortical carcinoma: clinical presentation and therapeutic strategies. AB - Adrenocortical cancers are relatively rare endocrine tumors that usually present when hormonally active or after they have become large and metastasis has occurred. Consequently, the 5-year survival rate is 20% to 35%. Surgical removal remains the only form of therapy proven to prolong survival. Mitotane is the most accepted form of chemotherapy. For the approximately 20% to 25% of patients whose tumors respond to mitotane, survival is prolonged. PMID- 9735135 TI - Surgical approach to adrenal neoplasms: laparoscopic versus open adrenalectomy. AB - There are a number of different approaches available to surgically remove the adrenal gland. These can be broadly classified into two general categories, open and laparoscopic. There is no one best method for all patients. Surgeons, skilled in all aspects of adrenal surgery, should choose an approach for adrenalectomy based on patient and tumor related factors. Once advantages and disadvantages of a specific procedure, with these factors in mind, are weighed, the best approach for the individual patient should be chosen. PMID- 9735136 TI - Insulinoma. AB - Symptoms most characteristically diagnostic of insulinoma are those of neuroglycopenia. The combination of hypoglycemia and endogenous hyperinsulinemia are pathognomonic of insulinoma. Several localization techniques are available, the choice of which best depends on the best expertise at individual institutions. Intraoperative ultrasonography is helpful in localization and defining related anatomy. Enucleation of these intrapancreatic tumors is preferred, but for body and tail lesions, distal pancreatic resection may be required. Because at least 90% are benign, long-term cure with complete resolution of preoperative symptoms is expected. PMID- 9735137 TI - Gastrinoma: advances in localization and treatment. AB - Gastrinomas secrete gastrin and cause symptoms related to gastric acid hypersecretion that can be controlled by antisecretory medications. Primary tumors are located within the pancreas or duodenum and 60% metastasize. Liver metastases are associated with decreased survival. Localization studies especially somatostatin receptor scintigraphy are indicated to image the extent of disease. Surgery is indicated to potentially cure the patient, or control the malignant tumoral process and prolong survival. PMID- 9735138 TI - Therapeutic alternatives in metastatic neuroendocrine tumors. AB - In the treatment of neuroendocrine tumors that cannot be resected for cure two goals must be addressed: (1) the control of symptoms related to hormonal hypersecretion and (2) the prolongation of survival by destruction of tumor of the limitation of its growth. Clinical approaches, medical and surgical, have been developed in recent years to meet these goals. Surgical options in the face of metastatic disease include resection of hepatic metastases, cryoablation, and liver transplantation. Medical therapy includes treatment of symptomatic hypersecretory states and systemic chemotherapeutic and immunomodulatory regimes. Hepatic artery chemoembolization has also been used successfully in the treatment of hepatic metastases of neuroendocrine tumors. PMID- 9735139 TI - Management of pancreatic endocrine tumors in patients with multiple endocrine neoplasia type 1. AB - The rationale for a multifaceted operative procedure in all MEN 1 patients with pancreaticoduodenal neuroendocrine disease who present without liver metastases is presented. The results in 36 patients with MEN 1 ZES are encouraging in that more than two-thirds are eugastrinemic and none have liver metastases after follow-up as long as 20 years. PMID- 9735140 TI - Management of follicular and Hurthle cell neoplasms of the thyroid gland. AB - Management of follicular and Hurthle cell neoplasms of the thyroid gland is a common clinical problem. A diagnosis of follicular or Hurthle cell carcinoma cannot be made from a fine-needle aspiration biopsy alone because it requires histologic demonstration of capsular or vascular invasion. Thyroid lobectomy and isthmusectomy is adequate treatment of benign follicular or Hurthle cell adenoma and minimally invasive follicular carcinoma. Total thyroidectomy, radioiodine ablation, and thyrotropin-suppressive doses of thyroid hormone is advocated for the invasive subtype of follicular carcinoma and all Hurthle cell carcinomas. Monitoring of serum thyroglobulin levels postoperatively is important for detection of recurrent disease. PMID- 9735141 TI - Analysis of 2-chloro-2'-deoxyadenosine incorporation into cellular DNA by quantitative polymerase chain reaction. AB - Thermus aquaticus (Taq) DNA polymerase elongation is blocked by several DNA adducts. This property has been exploited in polymerase chain reaction (PCR) methods to analyze cellular DNA damage and repair after exposure to damaging agents. Such methods have not been applied previously to detect nucleoside analog incorporation into cellular DNA. 2-Chloro-2'-deoxyadenosine (CldAdo), a deoxyadenosine analog, is clinically effective for hairy cell leukemia. CldAdo is taken up by cells, converted to the triphosphate, and incorporated into cellular DNA. Here, we measured by primer extension the ability of CldAMP residues in 98 base single-stranded DNA to block Taq elongation. In contrast to control DNA, no full-length 98-mers were produced on CldAMP-containing templates, and Taq polymerase was halted at the first CldAMP site. We then examined the possibility of using quantitative PCR to measure CldATP incorporation into the N-ras gene after incubation of cultured human leukemia cells with CldAdo or with cisplatin as a positive control for DNA damage. Treatment with either drug resulted in reduced amounts of amplified DNA product compared to untreated cells. CldAMP residues within cellular DNA inhibited PCR amplification in a dose-dependent manner; 100 nM CldAdo produced approximately 0.4 CldAMP sites within a 523-bp region of the N-ras sequence. Thus, PCR analysis with Taq polymerase provides a sensitive assessment of nucleoside analog incorporation after cellular exposure to antileukemic drugs. PMID- 9735142 TI - A method for assessing strand breaks in DNA. AB - A simple method has been developed to assess strand breaks in extracted DNA. The method uses the enzyme terminal deoxynucleotidyl transferase (TDT) to incorporate labeled deoxycytidine triphosphate (dCTP) in the presence of dideoxy-CTP (ddCTP) which is added to ensure that the reaction goes to completion. Following development of the method, the extent of DNA degradation in 21 blood or bone marrow samples, which had varying degrees of DNA degradation, was measured by the TDT assay, by gel electrophoresis, or by a laborious PCR-based method which quantifies the number of amplifiable N-ras targets in a sample. The TDT assay was more sensitive at detecting strand breaks than electrophoresis and there was good correlation between the results of the TDT assay and the N-ras assay. The TDT assay was also used to demonstrate the development of strand breaks during induced apoptosis. The TDT assay is thus a simple and semiquantitative method to study strand breaks produced by DNA damage. PMID- 9735143 TI - Short-term insulin-induced glycogen formation in primary hepatocytes as a screening bioassay for insulin action. AB - We describe a novel bioassay to measure specific insulin-like activity in primary cultures of rat hepatocytes by determination of [3H]glycogen from d-[6 3H]glucose. The dose-response curve of insulin in this assay exhibited an EC50 of 0.42 (+/-0.04) nM, which is comparable to the dissociation constant of insulin from its receptor in hepatocytes. We used this assay to examine possible residual insulin-like activity of the four major fragments formed upon insulin degradation by insulin protease. Fragments A1-13B1-9, A1-14B1-9,and A14-21B14-30 showed no measurable activity. Although preparations of fragment A14-21B10-30 displayed dose-dependent agonist activity with an EC50 of 380 (+/-40) nM, we conclude that this was due to an insulin-like impurity since the chemically synthesized fragment showed no such activity. In summary, this bioassay demonstrates the action of insulin on glycogen formation in hepatocytes and provides a rapid and sensitive measurement of insulin-like activity which could facilitate screening studies. PMID- 9735144 TI - Occurrence of oxidized metabolites of arachidonic acid esterified to phospholipids in murine lung tissue. AB - Isolation and characterization of murine pulmonary phospholipids revealed the normal occurrence of 10 isobaric eicosanoids corresponding to the incorporation of one oxygen atom into the arachidonate esterified to glycerophospholipids. Lungs from mice were removed and lipids were extracted and then separated into free carboxylic acid and phospholipids. Phospholipids were hydrolyzed to yield the free carboxylic acids prior to analysis. Reverse-phase HPLC and electrospray tandem mass spectrometry were used to identify and quantitate six monohydroxyeicosatetraenoic (HETE) and four epoxyeicosatetraenoic (EET) acid regioisomers using d8-HETE as internal standard. HETEs esterified to phospholipids were found to increase following intratracheal administration of tBuOOH (36 mg/kg), but not the levels of esterified EETs. Chiral analysis of esterified 15-HETE revealed an R/S ratio of 0.96, suggesting operation of a free radical mechanism responsible for generation of this monohydroxy arachidonate phospholipid, and this enantiomeric ratio was 1.10 following treatment of the mouse lung with tBuOOH. These results are consistent with a free-radical-based mechanism of oxidation of pulmonary glycerophospholipids containing arachidonate. PMID- 9735145 TI - Determination of ATP and its metabolites released from rat caudal artery by isocratic ion-pair reversed-phase high-performance liquid chromatography. AB - A sensitive and selective assay method for adenine compounds (purines) using high performance liquid chromatography with fluorescent detection was developed. The 1,N6-ethenoderivatives of adenine, adenosine, AMP, ADP, and ATP formed by reaction with chloroacetaldehyde at 80 degreesC were separated by ion-pair reversed-phase chromatography within 15 min under isocratic conditions. alpha,beta-Methylene adenosine 5'-diphosphate could be used as an internal standard for the determination of purines. The calibration graphs constructed with peak area ratios against amounts were linear between 0.1 and 10.0 pmol of each purine. The repeatability and intermediate precision were less than 6% (RSD, n = 5) and 8% (RSD, n = 3), respectively, at 0.5 pmol of each purine. A method for calculation of each purine amount which considers hydrolysis by derivatization is described. The optimized method was applied to determine the purines released from the rat caudal artery stimulated by noradrenaline. PMID- 9735146 TI - Development of an assay for the measurement of the surfactant pluronic F-68 in mammalian cell culture medium. AB - A colorimetric assay is described for the measurement of Pluronic F-68 in animal cell culture medium. The assay is based on the formation of a complex with cobalt thiocyanate as previously developed for the measurement of Pluronic in liver tissue. To adapt the assay for the lower detection levels required in cell culture medium, the absorbance of the complex was measured at 328 nm. The reproducibility of the assay was improved by washing the complex with ethyl acetate. The addition of a critical volume of ethanol was found to be necessary to reduce the interference caused by unknown components of the culture medium. The assay was linear between concentrations of 0.01 and 0.2% (w/v) Pluronic in serum-free medium. At the lower level of detection of 0.01% (w/v), the coefficient of determination (R2) was 0.998. The presence of serum in the medium decreased the sensitivity of the assay, which nevertheless was linear from 0.04 to 0.16% (w/v) Pluronic. The sensitivity and precision of the method are appropriate to study the dynamics of Pluronic in large-scale cell cultures in which Pluronic is added to reduce hydrodynamic cell damage. PMID- 9735147 TI - Total synthesis of 17,17,18,18-d4-iPF2alpha-VI and quantification of iPF2alpha-VI in human urine by gas chromatography/mass spectrometry. AB - Isoprostanes are a new class of natural products formed in humans as a result of free-radical-catalyzed lipid peroxidation of polyunsaturated fatty acids. These endogenous compounds are isomeric with biologically active prostaglandins and have great promise as markers of oxidant stress in vivo. iPF2alpha-III (previously 8-iso-PGF2alpha), an isoprostane from Class III (previously known as Class IV), has been used as an index of free-radical-induced oxidative stress. This isoprostane is also produced by the cyclooxygenase enzymes COX1 and COX2. We are proposing a new reliable index of oxidative stress based on iPF2alpha-VI (previously IPF2alpha-I), a new Class VI isoprostane we recently discovered. The advantages of iPF2alpha-VI are that it is several fold more abundant in urine than iPF2alpha-III, hence allowing more accurate determinations. Equally, the proximity of the C-5 OH function to the carboxylic acid allows the formation of the lactone 35 which is easier to purify from other iPs which cannot form such lactones. We have performed the first total synthesis of d4-iPF2alpha-VI by using two synthons, (3,3,4,4-d4)-hexylphosphonium bromide 23 prepared from 5-hexynol and syn-anti-syn lactone 25 synthesized from d-glucose. We have developed two variants of a sensitive GC/MS assay using the synthetic d4-iPF2alpha-VI as an internal standard to determine the levels of endogenous iPF2alpha-VI in biological fluids. Quantification of iPF2alpha-VI formed in vivo may be a more reliable index to assess oxidant stress in humans. PMID- 9735148 TI - A flow cytometric assay enabling specific detection of the human lysosomal enzyme, beta-glucocerebrosidase. AB - We have developed a flow cytometric assay specific for human lysosomal beta glucocerebrosidase (hGC) which is the enzyme deficient in Gaucher disease, a lysosomal storage disorder. The assay is based on the primate-specific monoclonal antibody 8E4 and thus allows detection of endogenous hGC and primate GC protein at a single cell level. We demonstrate that detection of endogenous hGC is possible in rhesus and human cells. Since antibody 8E4 does not bind to rodent GC, hGC detection in murine cell lines and primary cells upon transduction with a retrovirus carrying the hGC cDNA is possible. Comparison of this assay to a flow cytometric method which detects enzymatic GC activity shows that the 8E4-based assay is significantly more sensitive. We also show that multiparameter analyses in combination with hGC detection are feasible. This enables hGC detection in different lineages of complex cell populations. The increased sensitivity in combination with the specificity for hGC makes the 8E4-based flow cytometric assay ideally suited to monitor hGC expression. This assay is therefore of significant value to monitor the success of therapeutic strategies for Gaucher disease such as enzyme supplementation therapy, allogeneic bone marrow transplantation, and gene therapy. PMID- 9735149 TI - Probing RegA/RNA interactions using electrospray ionization-fourier transform ion cyclotron resonance-mass spectrometry. AB - The interactions of bacteriophage T4 regA protein, a unique translational regulator, with RNAs of various size and sequence were studied using electrospray ionization-Fourier transform ion cyclotron resonance-mass spectrometry. Using very gentle interface conditions, regA/RNA complexes with a 1:1 binding stoichiometry were observed for all four target RNAs studied, consistent with solution binding studies. Competitive binding of target RNAs and their degradation products with regA demonstrated that the loss of a single nucleotide resulted in a dramatic change in binding affinity in some cases. Competitive binding of regA with four target RNAs revealed similar relative binding affinity order to that suggested by previous in vitro repression experiments. The use of sustained off-resonance irradiation for collisionally induced dissociation of a regA/RNA complex suggested the potential for directly obtaining information regarding the regA binding domain. PMID- 9735150 TI - Detection of rare target genes on northern blots with cDNA probes labeled by reverse transcriptase-polymerase chain reaction with simultaneous digoxigenin incorporation. PMID- 9735151 TI - DNA isolation from recalcitrant materials such as tree roots, bark, and forest soil for the detection of fungal pathogens by polymerase chain reaction. PMID- 9735152 TI - Concentration and desalting of protein samples for mass spectrometry analysis. PMID- 9735153 TI - Measurement of DNA in intervertebral disc and other autofluorescent cartilages using the dye hoechst 33258. PMID- 9735154 TI - Peptide substrates dissolved in dimethylformamide may be modified at the epsilon amino group of lysyl residues, causing erroneous kinetic characterization of proteolytic enzymes. PMID- 9735155 TI - Analysis of protein-protein interaction by two-dimensional affinity electrophoresis. PMID- 9735156 TI - Isolation of plasmid DNA using magnetite as a solid-phase adsorbent. PMID- 9735157 TI - A rare case of false positive in a yeast two-hybrid screening: the selection of rearranged bait constructs that produce a functional gal4 activity. PMID- 9735158 TI - Ritodrine inhibition of the plasma membrane Ca2+-ATPase from human erythrocyte. AB - The Ca(2+)-ATPase activity of human erythrocyte membrane can be inhibited in vitro by ritodrine, a beta 2-adrenergic agonist. The inhibitory profile shows a low-affinity interaction and no competition with the specific transport and catalytic substrates. The activated conformation of the enzyme (in the presence of calmodulin or after trypsin digestion) facilitates the interaction with ritodrine. This suggests that the C-terminal tail of the enzyme plays a protective role. By studying selected partial reactions of the catalytic and transport cycle we found that the inhibition can be basically assigned to a lower rate of phosphorylation by ATP. A minor effect on the phosphorylation level by Pi in the absence of Ca2+ and no effect on the enzyme affinity for Ca2+ or ATP were also observed. The inhibition of the plasma membrane Ca(2+)-ATPase by ritodrine shows a clear similarity with that of the sarcoplasmic/endoplasmic reticulum membrane. The inhibition under study does not foresee a pharmacological effect of ritodrine on the myometrial plasma membrane Ca(2+)-ATPase when administered for the management of preterm labor. PMID- 9735159 TI - Inhibition of liposome-induced complement activation by incorporated poly(ethylene glycol)-lipids. AB - Complement activation causes opsonization of foreign particles leading to particle elimination from the blood. Complement-mediated opsonization of charged and large liposomes presents a fundamental problem in their use to deliver therapeutic agents in vivo. To prolong the circulation half-lives of such liposomes, complement activation must be curtailed. The aim of this study was to assess the ability of poly(ethylene glycol)-lipids (PEG-lipids) to inhibit the in vitro activation of the classical pathway of complement in human serum by anionic liposomes. Incorporation of cholesterol-PEG600 (CH-PEG600), cholesterol-PEG1000 (CH-PEG1000), or phosphatidylethanolamine-PEG2000 (PE-PEG2000) resulted in dose dependent inhibition of C1q binding and complement activation. The dose of PEG lipid at which complement activation was blocked was inversely related to the PEG chain length. Complement activation was strongly inhibited when 15 mole% of CH PEG600, 10 mole% CH-PEG1000, or 5 mole% PE-PEG2000 was incorporated into 100-nm anionic liposomes. PEG-lipid incorporation into larger liposomes (240 nm) was also successful in blocking C1q binding and complement activation. Radiolabeled cholesterol-PEG approximately 1400 was prepared and used to determine both the percentage of CH-PEG incorporated into the liposomes and the percentage maintained in the liposomes in the presence of 50% human serum at 37 degrees C for up to 24 h. PMID- 9735160 TI - Neuronal nitric oxide synthase is refractory to mechanism-based inactivation in GH3 pituitary cells. AB - Nitric oxide formation by GH3 pituitary cells is stimulated by depolarizing concentrations of K+ and by the L-channel Ca2+ agonist Bay kappa 8644 in an additive manner that depends on extracellular Ca2+. Ca(2+)-dependent NO formation at 100 microM arginine was inhibited 50% over a 30-min period by 5 microM NG amino-L-arginine, 30 microM N6-iminoethyl-L-ornithine (NIO) and 520 microM N5 iminoethyl-L-lysine (NIL) but required concentrations of aminoguanidine (AG) greater than 3 mM. As measured at 100 microM extracellular arginine, intracellular neuronal nitric oxide synthase (nNOS) was inactivated 50% over a 30 min period by 150 microM NG-amino-L-arginine and 1500 microM NIO, but required concentrations of NIL or AG greater than 5 mM. The inactivation of nNOS by these agents occurred only under conditions that mobilized extracellular Ca2+ and was inhibited by increased extracellular arginine. Thus these agents inhibit cellular Ca(2+)-dependent NO formation at concentrations far lower than those required to inactivate the cellular nNOS. Inhibition of NO formation by these agents was not attributable to effects on cellular arginine uptake. In contrast diphenyliodonium chloride produced a rapid and complete inactivation of cellular NO formation and nNOS activity. These inactivations produced by diphenyliodonium chloride occurred with identical kinetics but displayed no requirement for Ca2+. These data support the assertion that neuronal NO synthase is refractory to mechanism-based inactivation in GH3 pituitary cells. PMID- 9735161 TI - A simple procedure for purifying the major chloroplast fructose-1,6 bisphosphatase from spinach (Spinacia oleracea) and characterization of its stimulation by sub-femtomolar mercuric ions. AB - A rapid procedure for the purification of the redox-regulated chloroplast fructose-1,6-bisphosphatase [EC 3.1.3.11] from spinach leaf extract to homogeneity is described. No thiol-reducing agents were present during the purification and the enzyme is > 99% in the oxidized form. A rapid procedure to reduce and activate the Fru-1,6-P2ase by dithiothreitol in the absence of thioredoxin is described. Reduction activates the enzyme up to several hundred fold when assayed at pH 8.0 with 2 mM Mg2+. The activity of the purified oxidized enzyme is unusually sensitive to changes in Mg2+ and H+ concentration. Tenfold changes in Mg2+ or H+ concentration lead to > 100-fold increases in activity. The recoveries of fructose-1,6-bisphosphatase activity as determined by the activity of the oxidized enzyme at pH 8.0/20 mM Mg2+; pH 9.0/2 mM Mg2+; pH 8/2 mM Mg2+ plus 0.1 mM Hg(II) or of the reduced enzyme at pH 8.0/2 mM Mg2+ are similar (approximately 40%) indicating that the major proportion of these activities in a leaf extract is catalyzed by the same enzyme. Moreover, antibodies raised against the purified enzyme inhibit all of the above activities in crude leaf extracts. The kinetic properties of the purified enzyme suggest that the oxidized Mg(2+) dependent enzyme can play no significant role in photosynthetic carbon assimilation. A survey of some kinetic properties of Fru-1,6-P2ase activity in extracts of various photosynthetic organisms reveals that all 11 species examined possess a redox- and pH/Mg(2+)-stimulated Fru-1,6-P2ase, whereas Fru-1,6-P2ase in extracts of Taxus baccata (a gymnosperm), Chlorella vulgaris (a green alga), and the cyanobacterium Nostoc muscorum were not activated by Hg(II). The heat stability that proved useful in the purification of the spinach enzyme was conserved in both angiosperms and gymnosperms. The oxidized enzyme (which normally has no thiol groups accessible to 5,5'-dithio-bis[2-nitrobenzoic acid]) but not the reduced enzyme can be stimulated many hundred-fold by addition of extraordinarily low concentrations of Hg(II) to a complete assay mixture. With the aid of EDTA as a Hg(II) buffer, half-maximal stimulation was achieved at 2 x 10(-16) M free Hg(II). Methylmercury also stimulates the enzyme many hundred-fold at very low concentrations. The concentration for half-maximal stimulation by methylmercury was determined with a cyanide buffer to be approximately 10(-16) M. This, together with the high affinity of the enzyme for Hg(II), suggests that Hg(II) stimulates the enzyme by binding to an enzyme thiol group that be comes exposed in the catalytically active enzyme, thereby stabilizing the oxidized enzyme in an active conformation. By contrast, in the absence of Fru-1,6-P2 and either Mg2+ or Ca2+, Hg(II) (even at 2 x 10(-16) M) rapidly inactivates the oxidized Fru-1,6-P2ase. This inactivation is similar to the inactivation of Fru 1,6-P2ase that occurred at high pH (> 9) and which is also prevented by Fru-1,6 P2 and either Mg2+ or Ca2+. Although the Hg(II)- and high pH-inactivated oxidized enzyme has no activity, both forms of the enzyme can be activated by reduction. The usefulness of buffers to maintain low, defined Hg(II) and organic mercurial concentrations is discussed. PMID- 9735162 TI - Suppression of copper-induced cellular damage by copper sequestration with S100b protein. AB - We previously reported that S100b protein (homodimer of S100 beta subunit) can bind copper ions with a submicromolar dissociation constant (T. Nishikawa et al., J. Biol. Chem. 272, 23037-23041, 1997). In this study, a question was addressed as to whether this protein can sequester copper ions in an in vivo situation. Escherichia coli cells that had been rendered able to produce a fusion protein of rat S100 beta subunit with glutathione S-transferase displayed a marked resistance to cellular damage induced by copper alone or its combination with H2O2, compared with control cells expressing the transferase moiety only. A study by gel chromatography showed that about half of the expressed S100 beta fusion protein in the cytosol of copper-treated cells was eluted in the void volume fraction (molecular mass > 200 kDa), which contained most of the copper incorporated. The S100 beta fusion protein purified from the void volume fraction was found to contain 82% of the total copper in the fraction, while in a parallel experiment with the control cells, the glutathione S-transferase eluted in the void volume fraction contained only 18% of the total copper. Thus, it is clear that extraneously expressed S100b protein can acts as a "copper sink," thereby protecting E. coli cells from copper-induced cellular damage. PMID- 9735163 TI - Reactive oxygen species generated from the reaction of copper(II) complexes with biological reductants cause DNA strand scission. AB - The generation of hydroxyl radicals (.OH) from the reaction of Cu(II) complexes with biological reductants such as ascorbic acid, glutathione, acetylcysteine, and hydroquinone was confirmed by spin-trapping experiments using electron spin resonance (ESR). The following Cu(II) complexes were used: Cu(II)-(CyHH)2 (CyHH, cyclo(L-histidyl-L-histidyl)), Cu(II)(OP)2 (OP, o-phenanthroline), Cu(II)(HGG) (HGG, L-histidyl-glycylglycine), and Cu(II)(en)2 (en, ethylenediamine). The methyl radical adduct of alpha-(pyridyl-4-N-oxide)-N-tert-butylnitrone (POBN-CH3) was obtained from the reaction of ascorbic acid with all Cu(II) complexes used here in the presence of a spin trap, POBN, and dimethyl sulfoxide, indicating the generation of .OH. Glutathione, N-acetylcysteine, and hydroquinone reacted with both Cu(II)(CyHH)2 and Cu(II)(OP)2 to generate POBN-CH3, while these reductants did not react with either Cu(II)(HGG) or Cu(II)(en)2. Interestingly, the formation of POBN-CH3 in the reaction of Cu(II)-(CyHH)2 with glutathione or N acetylcysteine was found only at a Cu(II)(CyHH)2/glutathione or Cu(II)(CyHH)2/N acetylcysteine ratio of 1. The DNA strand scission caused by reaction mixtures of Cu(II) complexes with reductants was investigated under the same conditions as the ESR spin-trapping experiments. Addition of ascorbic acid to mixtures of these four Cu(II) complexes and DNA resulted in DNA strand breakage. Hydroquinone plus Cu(II)(CyHH)2 also caused DNA strand scission. In addition, DNA strand breakage was observed with the reaction of Cu(II)(OP)2 with glutathione, N-acetylcysteine, and hydroquinone. In contrast, reaction mixtures of glutathione, N acetylcysteine, or hydroquinone with Cu(II)-(HGG) or Cu(II)(en)2 did not cause DNA strand scission within the concentration range used. The results obtained here suggest that there is a good correlation between POBN-CH3 formation and DNA strand scission. Thus, DNA strand scission may be caused by .OH generated from the reaction of some Cu(II) complexes with biological reductants under aerobic conditions. Since ascorbic acid, glutathione, and N-acetylcysteine are present in living cells, some Cu(II) complexes may be capable of initiating DNA damage in the presence of these reductants. PMID- 9735164 TI - Identification of residues 286 and 289 as critical for conferring substrate specificity of human CYP2C9 for diclofenac and ibuprofen. AB - Specificity of human CYP2C9 for two substrates, diclofenac and ibuprofen, was studied using chimeras and site-directed mutants of CYP2C9 and the highly related CYP2C19 expressed in Escherichia coli. Data were correlated with the presence of putative substrate recognition sites (SRS). A CYP2C19 chimera containing residues 228-340 (SRS 3 and 4) of 2C9 conferred both diclofenac hydroxylation and 2- and 3 hydroxylation of ibuprofen. The regiospecificity of this construct for metabolism of ibuprofen differed from that of CYP2C9 by favoring 2-hydroxylation over 3 hydroxylation. A CYP2C9 construct containing residues 228-340 of CYP2C19 lacked both diclofenac and ibuprofen hydroxylase activities. When residues 228-282 (containing SRS 3) of CYP2C9 were replaced by those of CYP2C19, the chimera retained appreciable activity for diclofenac and ibuprofen, and tolbutamide activity was inhibited by a specific CYP2C9 inhibitor, sulfaphenazole. This suggested that SRS 3 is not important in conferring specificity. CYP2C9 and CYP2C19 differ in five residues within the region 283-340 (within SRS 4). Mutations to analyze SRS 4 were made on a CYP2C19 chimera containing residues 228 282 of CYP2C9. A single I289N mutation conferred a dramatic increase in diclofenac hydroxylation and a small increase in ibuprofen 2-hydroxylation. A second mutation (N286S and I289N) increased diclofenac hydroxylation and conferred a dramatic increase in ibuprofen 2-hydroxylation. A V288E mutation did not increase activity toward either substrate and decreased activity toward the two substrates in combination with the I289N or the N286S, I289N mutants. Therefore residues 286 and 289 of CYP2C9 are important in conferring specificity for diclofenac and ibuprofen. PMID- 9735165 TI - Mutagenesis disrupts posttranslational processing of the Na,K-ATPase catalytic subunit. AB - The first 5 amino acids of the catalytic alpha 1 isoform from Na,K-ATPase are cleaved enzymatically during or after translation. To evaluate the structural requirements for that cleavage, we constructed amino-terminal mutants of alpha 1 in which an epitope tag from the c-myc oncogene product was added. Immunoblots of isolated membranes from transfected monkey kidney cells revealed binding of an antibody specific for the first 9 residues of the alpha 1 nascent protein. Because this antibody does not recognize the shorter sequence corresponding to the processed polypeptide, these results indicate that the epitope tag prevented normal processing, a conclusion confirmed by the observed binding of an anti-myc antibody. In contrast, membranes from cells expressing deletion mutants that lack residues 10-24 and 10-31 of the nascent chain failed to bind the amino-terminal directed antibody, suggesting that the mutants were cleaved normally and that amino acids downstream of the first 9 are not required for proteolysis. Amino terminal mutants produced in other laboratories have shown an anomalous stimulation of ATPase activity by K+ when measured in low ATP concentrations. The myc-tagged and downstream deletion mutants were sensitive to K+ in the range from 0.05 to 5 mM, similar to wild-type enzyme, despite the differences in posttranslational processing. A mutant missing the first 40 residues of the nascent chain, however, displayed an activation by K+. These results suggest that amino-terminal processing of the alpha 1 isoform was prevented by mutation, yet that processing had little influence on the kinetic parameter most likely to be influenced by such changes. PMID- 9735166 TI - Effects of 15-oxa-32-vinyl-lanost-8-ene-3 beta,32 diol on the expression of 3 hydroxy-3-methylglutaryl coenzyme A reductase and low density lipoprotein receptor in rat liver. AB - The mechanisms by which oxylanosterols regulate expression of hepatic 3-hydroxy-3 methylglutaryl coenzyme A (HMG-CoA) reductase and lower serum cholesterol levels were examined by using a novel nonmetabolizable oxylanosterol mimic, 15-oxa-32 vinyl-lanost-8-ene-3 beta, 32 diol (DMP 565). This compound, unlike other nonmetabolizable oxylanosterols, is not a substrate for lanosterol 14 alpha methyl demethylase. Feeding rats a diet supplemented with 0.02% DMP 565 markedly decreased HMG-CoA reductase immunoreactive protein and enzyme activity levels without affecting mRNA levels. The rate of reductase protein degradation was unaffected. However, the rate of translation was reduced to less than 20% of control. Thus, DMP 565 appears to regulate hepatic HMG-CoA reductase gene expression primarily at the level of translation. The pronounced inhibition of HMG-CoA reductase by DMP 565 resulted in a compensatory increase in the functioning of the hepatic low density lipoprotein (LDL) receptor, possibly by increased cycling, as evidenced by a marked increase in the rate of degradation of the LDL receptor. The half-life of the receptor was decreased from over 7 h to only 1 h in animals receiving DMP 565. This increase in the rate of degradation occurred without a change in the steady state level of the receptor. Addition of dietary cholesterol attenuated the increased turnover of the LDL receptor. These effects on the hepatic LDL receptor have also been observed with HMG-CoA reductase inhibitors (G. C. Ness et al., 1996, Arch. Biochem, Biophys. 325, 242 248). However, the effect of DMP 565 on the rate of degradation of the hepatic LDL receptor was of a greater magnitude when equal doses of the drugs were used. These regulatory actions of DMP 565 provide, in part, an explanation for the observed hypocholesterolemic action of this compound. PMID- 9735167 TI - Reactions of hypochlorous acid with biological substrates are activated catalytically by tertiary amines. AB - The activation of reactions of HOCl with a variety of model substrates by tertiary amines was investigated spectroscopically by tandem-mix and stopped-flow techniques. HOCl-induced chlorination of salicylate can be sped up by several orders of magnitude by catalytic amounts of trimethylamine (TMN). The effect is obviously due to the fast generation of reactive quarternary chloramonium ions, TMN+ Cl, which act as chain carrier in a catalytic reaction cycle. Of various catalysts tested, quinine shows the highest activity; this is attributable to the quinuclidine (QN) substituent, a bicyclic tertiary amine, forming a particularly reactive chloro derivative, QN+ Cl, which does not decompose autocatalytically. The rate of catalytic salicylate chlorination as a function of pH (around pH 7) depends not at least on the basicity of the tertiary amine; the rate increases with pH in the cases of TMN and quinuclidine (high basicity), but decreases with pH in the case of MES (low basicity). Tertiary amines also catalyze the interaction between HOCl and alkenes, as shown using sorbate as model. Reaction of HOCl with the nucleotides GMP and CMP is sped up remarkably by catalytic amounts of tertiary amines. In the case of GMP the same product spectrum is produced by HOCl in absence and presence of catalyst, but a change in the product spectra is obtained when AMP and CMP are reacted with HOCl in presence of catalyst. Using poly(dA-dT).poly(dA-dT) as DNA model, it is shown that HOCl primarily induces an absorbance increase at 263 nm, which indicates unfolding of the double strand due to fast chlorination of thymidine; a subsequent secondary absorbance decrease can be explained by slow chlorination of adenosine. Both the primary and secondary processes are activated by catalytic amounts of quinine. No evidence was found for a radical pathway in TMN-mediated oxidation of formate by HOCl. The present results suggest that low concentrations of certain tertiary amines have the potential of modifying the spectrum of target molecules which can be damaged by HOCl in biological systems. PMID- 9735168 TI - Alcohol-induced molten globule intermediates of proteins: are they real folding intermediates or off pathway products? AB - Alcohols have been shown to cause a conformational transition of proteins into a new stable conformational state resembling that of the "molten globule intermediate" characterized by high alpha-helical content and disrupted tertiary structure. We have studied the effect of monohydric alcohols on the stability and structural characteristics of small globular protein hen egg white lysozyme by the combined use of differential scanning calorimetry, circular dichroism, and nuclear magnetic resonance spectroscopy. The protein stability was found to be significantly decreased with increasing alcohol concentration, and, in presence of moderate to higher alcohol concentrations, depending on the pH and alcohol studied, the protein was found to be unfolded even at 4 degrees C. Correlation between thermal stability and alpha-helicity of several small globular proteins like hen egg white lysozyme, horse heart cytochrome C, and bovine carbonic anhydrase B, observed in presence of increasing alcohol concentrations, suggests that probably alcohols induce helical structures in unfolded protein. The temperature-dependent near- and far-UV circular dichroism and proton nuclear magnetic resonance spectroscopic studies on lysozyme in the presence of 2,2,2 trifluoroethanol and methanol, respectively, showed that alcohols do induce significantly higher helical structures in unfolded protein compared to folded protein. The results presented in this paper suggest that the molten globule intermediate of proteins in the presence of high alcohols as reported earlier is due to alcohol-induced local folding rather than global folding of unfolded protein and hence is an off-pathway product and not a real folding intermediate. PMID- 9735169 TI - Evaluation of phosphoenolpyruvate as a phosphoryl group donor for phosphoproteins in skeletal muscle. AB - The possible existence of a phosphoenolpyruvate-dependent protein kinase activity in muscle first reported by Khandelwal et al. (FEBS Lett. 162, 127-132, 1983) was further examined in this study. [32P]Phosphoenolpyruvate was used to assay rabbit muscle extract that was first dialyzed and then passed over a gel filtration column. Fractions from the column were assayed for activity (as shown by counts incorporated into an acid-precipitable product), which was observed only when two distinct and resolved fractions were combined. The approximate masses of these two components, as estimated by nondenaturing gel filtration chromatography, were 30 and 160 kDa. The activity showed time and concentration dependence and was inactivated by heat and proteolysis. [gamma-32P]ATP could not substitute for [32P]phosphoenolpyruvate in the activity assays, nor was there evidence for the formation of ATP during the assays. Elution of the reaction mixture from a sizing column revealed several radioactive peaks. Other tissues (heart, kidney, liver, lung, spleen, and back skeletal muscle) from rabbit and rat were screened; the total activity was detected in all tissues examined, but was highest in the skeletal muscle of both species, with that from rabbit being twice that from rat. PMID- 9735170 TI - Iron and phosphate content of rat ferritin heteropolymers. AB - An attempt was made to relate the iron and phosphate content of ferritin to its subunit composition. Ferritins from various tissues were separated according to their subunit composition by anion exchange chromatography and according to their iron content by density-gradient centrifugation. Iron and phosphate contents were not related to subunit composition. Recombinant rat liver ferritin heteropolymers of different subunit composition (1, 4, 6, 10, 15, and 17 H chains per 24 mer) were maximally loaded with iron, using ceruloplasmin and phosphate. All loaded approximately the same amount of iron and phosphate (2250 and 380 atoms, respectively). The iron and phosphate content of all ferritin, including the maximally loaded recombinant ferritin heteropolymers, fit an equation we previously reported: [Fe] = 4404 - 5.61 [Pi] (D. deSilva et al., 1993, Arch. Biochem. Biophys. 303, 451-455). These results suggest that the amount of iron and apparently the space within the core of ferritin were not related to different subunit composition. PMID- 9735171 TI - Phosphorylation of tau at both Thr 231 and Ser 262 is required for maximal inhibition of its binding to microtubules. AB - The paired helical filaments (PHFs) found in Alzheimer's disease (AD) brains are composed primarily of the microtubule-associated protein tau. PHF-tau is in a hyperphosphorylated state and is unable to promote microtubule assembly. We investigated whether the inhibition of tau binding to microtubules is increased when tau is phosphorylated by different kinases in combination with GSK-3. We found that when tau was first phosphorylated by A-kinase, C-kinase, cdk5, or CaM kinase II and then by GSK-3, its binding to microtubules was inhibited by 45, 61, 78, and 79%, respectively. Further, the kinase combinations cdk5/GSK-3 and CaM kinase II/GSK-3 rapidly phosphorylated the sites Thr 231 and Ser 235. When these sites were individually replaced by Ala and the phosphorylation experiments repeated, tau binding to microtubules was inhibited by 54 and 71%, respectively. By comparison, when Ser 262 was replaced by Ala, tau binding to microtubules was inhibited by only 8% after phosphorylation by CaM kinase II. From these observations we estimate that the phosphorylation of Thr 231, Ser 235, and Ser 262 contributes approximately 26, approximately 9, and approximately 33%, respectively, of the overall inhibition of tau binding to microtubules. Together, our results indicate that the binding of tau to microtubules is controlled by the phosphorylation of several sites, among which are Thr 231, Ser 235, and Ser 262. PMID- 9735172 TI - Purification and characterization of NADPH-cytochrome P450 reductase from filamentous fungus Rhizopus nigricans. AB - We report here the isolation and partial characterization of a flavoprotein, NADPH-cytochrome P450 (cytochrome c) reductase. The enzyme is a part of steroid 11 alpha-hydroxylating system and is associated with the microsomal fraction of the fungus Rhizopus nigricans. Fungal reductase was solubilized from microsomal membranes with Triton X-100 and purified to apparent homogeneity by affinity and high-performance ion-exchange chromatography. A 350-fold purification of the enzyme with specific activity of 37 mumol cytochrome c reduced/min/mg protein was achieved. A single protein band was obtained on SDS-PAGE analysis with an apparent molecular weight of 79 kDa. Purified reductase contained approximately equimolar quantities of flavin adenine dinucleotide and flavin mononucleotide per mole of the enzyme. Upon induction of the steroid hydroxylating system with progesterone the activity of microsomal NADPH-cytochrome c (P450) reductase increased 10-fold. This is in good correlation with the increase in content of fungal cytochrome P450. Purified fungal flavoprotein was active in a reconstituted system with cytochrome P450 C21 from adrenal gland but could not replace adrenodoxin reductase in the mitochondrial steroid 11 beta-hydroxylating system. We were able to confirm the role of the enzyme by reconstituting steroid 11 alpha-hydroxylating activity from the separated components NADPH-cytochrome P450 reductase and cytochrome P450, partly purified from fungal microsomes. PMID- 9735174 TI - A novel method for the purification of selenoprotein P from human plasma. AB - Selenoprotein P was purified from human plasma using conventional chromatographic methods featuring metal-chelate-affinity chromatography as the final step. Two distinct isoforms with different selenium content were isolated and identified by N-terminal sequencing and immunoblot analysis. Their molecular mass is 61 and 51 kDa, respectively. Both isoforms could be detected in fresh plasma from five individuals. This rules out the possibility of the second isoform being an artifact which results from degradation of full-length selenoprotein P during purification. PMID- 9735173 TI - The N-end rule pathway in Xenopus egg extracts. AB - Ubiquitin-dependent degradation of intracellular proteins underlies a multitude of biological processes, including the cell cycle, cell differentiation, and responses to stress. One ubiquitin-dependent proteolytic system is the N-end rule pathway, whose targets include proteins that bear destabilizing N-terminal residues. This pathway, which has been characterized only in somatic cells, is shown here to be present also in germ line cells such as the eggs of the amphibian Xenopus laevis. We demonstrate that the set of destabilizing residues in the N-end rule pathway of Xenopus eggs is similar, if not identical, to that of somatic cells such as mammalian reticulocytes and fibroblasts. It is also shown that the degradation of engineered N-end rule substrates in egg extracts can be strongly and selectively inhibited by dipeptides bearing destabilizing N terminal residues. This result allowed us to ask whether selective inhibition of the N-end rule pathway in egg extracts influences the apoptosis-like changes that are observed in these extracts. A dipeptide bearing a bulky hydrophobic (type 2) destabilizing N-terminal residue was found to delay the apoptotic changes in egg extracts, whereas dipeptides bearing basic (type 1) destabilizing N-terminal residues had no effect. High activity of the N-end rule pathway in egg extracts provides an alternative to reticulocyte extracts for the in vitro analyses of this pathway. PMID- 9735175 TI - Age-related differences in random generation. AB - This study investigated the effects of age on a random generation task. In Experiment 1, young and elderly subjects were asked to generate random strings of letters at 1-, 2-, and 4-s rates. The elderly subjects produced more alphabetical stereotype responses than young subjects, even in the slowest rate condition. Furthermore, as faster rates were imposed, elderly subjects could no longer maintain the pace and missed responses. In Experiment 2, subjects were required to generate letters at the same time that they sorted cards into one, two, four, or eight categories. Age-related differences were observed on most of the measures of randomness (stereotypes, zero-order, and first-order measures). In addition, the number of errors increased with the number of sorting alternatives, especially for elderly subjects. These results suggest the existence of a reduction of the central executive resources, along with a reduced inhibition ability, in the elderly subjects. However, the contribution of a perceptual speed factor is also discussed. PMID- 9735176 TI - Hemisphere asymmetry in sympathetic control of the human myocardium. AB - Hemisphere asymmetry in sympathetic control of myocardial performance was studied in healthy human subjects using lateralized film presentation for selective sensory stimulation of the hemispheres and impedance cardiography for the evaluation of cardiac output, systolic time intervals and myocardial contractility. Results revealed a clear and consistent right hemisphere predominance in sympathetically mediated control of various components of myocardial performance. There is reason to assume that the obtained hemisphere differences in autonomic control of the heart are self-reliant processes not depending on emotion-related hemisphere asymmetry. As far as we know, this is the first study examining the distinct roles of the cerebral hemispheres in neural control of ventricular myocardial functions. PMID- 9735177 TI - A study of emotional processing in Parkinson's disease. AB - This study investigated three aspects of processing materials with emotional content in patients with idiopathic Parkinson's disease (PD): the ability to produce affective prosody, to discriminate affectively loaded speech, and to detect the surprise element in humorous sketches. Study aims were the characterization of an emotional processing deficit, and to test whether impaired emotional processing is mental state dependent. Forty-eight nondemented PD patients were divided according to neuropsychological criteria into a sample with intact mental functions and a sample with mild to moderate cognitive deterioration, particularly memory impairment. PD patients with intact cognitive functions were solely impaired at producing affectively loaded sentences, but otherwise displayed normal emotional processing abilities as compared to a clinical control group. PD patients with mental impairment were significantly disabled on all three tasks. The observed emo tional processing deficit was not related to variables like age, disease duration, de gree of functional impairment, motor disability or depression. Active and receptive emotional prosody were significantly correlated. Further strong positive correlations were found between the ability to disclose pictorial humour and tasks of visuoconceptual knowledge, as well as between the ability to produce affectively loaded speech and years of schooling. These results were interpreted as indicating that not only the production of emotional prosody, but also its recognition and the discovery of pictorial humour are reduced in a subgroup of PD patients with mental impairment. Impaired emotional processing skills are mental state dependent findings in PD which seem to be independent from demographic or disease variables and may indicate beginning dementia. PMID- 9735178 TI - Hand-mouth coordination, congenital absence of limb, and evidence for innate body schemas. AB - Studies of phantom limb in cases of congenital (aplasic) absence of limb have provided inadequate evidence concerning the innate neurological substrate responsible for the phantom. In this study we review evidence from ultrasonic and behavioral studies of hand-mouth coordination in utero and in early infancy, neurobiological studies in primates, and studies of neural reorganization following amputation. We suggest two complementary hypotheses to explain aplasic phantoms. First, aplasic phantoms are based on the existence of specific neural circuitry associated with innate motor schemas, such as the neural matrix responsible for early hand-mouth coordination. Second, aplasic phantoms are modified by mechanisms that involve a reorganization of neural representations of the missing limb within a complex network involving both cortical and subcortical structures. PMID- 9735179 TI - The use of advance information for motor preparation in Parkinson's disease: effects of cueing and compatibility between warning and imperative stimuli. AB - The ability of 13 Parkinsonian patients and 11 age-matched control subjects to process and use two components of the information given prior to a voluntary movement was studied using reaction time (RT) tasks. This advance information about the direction of a pointing movement was given using a double stimulation paradigm with an auditory warning signal (WS) which occurred prior to a visual imperative signal (IS). The first component of the information was given by the WS at the beginning of each trial, and the second component was the WS-IS compatibility during series of trials. The subjects were tested with three RT paradigms: a cued simple (CS) task, a cued choice (NC) task, and a priming choice (P) task. The results show that the normal subjects used both the lateral cue and the WS-IS compatibility to shorten their RTs, whereas the Parkinsonian patients were able to use the lateral warning signal, but their ability to use the degree of compatibility stimuli was impaired. These data suggest that when dealing with lateral cues in a RT task, Parkinsonian patients have no difficulty in identifying a stimulus and selecting the appropriate response, but that this is no longer so in the case of stimulus compatibility. This impairment may be due to attentional disorders involving a dysfunction affecting the medial premotor system, which includes the basal ganglia and may be responsible for the feedforward movement control deficits associated with Parkinson's disease. PMID- 9735180 TI - Handedness and sex differences in intelligence: evidence from the medical college admission test. AB - Our analysis of Medical College Admission Test subtest scores by writing hand preference and sex suggests that (a) right hemispheric dominance is associated with intellectual giftedness in verbal reasoning (left-handers obtained higher scores on the verbal reasoning test and were overrepresented in the upper tail of the distribution), (b) different patterns of brain lateralization are associated with different subcomponents of cognition (right-handers scored higher, on average, on the writing test and were overrepresented in the upper tail of the distribution), and (c) men generally score higher than women on tests of scientific knowledge (the most striking differences between men and women were on the biological and physical science tests). PMID- 9735181 TI - Manual asymmetries in goal-directed movement: examination of the motor output hypothesis. AB - Two experiments are reported which examined the viability of motor output hypothesis as an explanation for manual asymmetries in goal-directed movement. Experiment 1 isolated the variability due to force generation by directly assessing precision of force production during an isometric wrist flexion task. Experiment 2 examined the additional role of externally based and internally created timing patterns on the performance of a repetitive force production task. Virtually no effects involving hand were apparent in either experiment. These findings provide no support for a hypothesis based solely on motor output to adequately account for hand differences in the performance of rapid, goal directed movement. PMID- 9735184 TI - Size Tailoring of Magnetite Particles Formed by Aqueous Precipitation: An Example of Thermodynamic Stability of Nanometric Oxide Particles. AB - The particle mean size of magnetite precipitated in aqueous solution can be adjusted and stabilized against ripening over a large range at the nanometric scale (1.5-12.5 nm). Such a tailoring of particles is obtained by controlling the pH and the ionic strength imposed by a noncomplexing salt in the precipitation medium. The higher the pH and the ionic strength are, the smaller the particle size is. Above a critical pH value, which depends on the ionic strength and the temperature, the secondary particle growth by Ostwald ripening does not take place anymore. The stabilization of nanoparticles seems to result from thermodynamics rather than kinetics. Copyright 1998 Academic Press. PMID- 9735185 TI - Theory and Experiment on the Measurement of Kinetic Rate Constants for Surfactant Exchange at an Air/Water Interface. AB - The paper focuses on the measurement of the rate constants for the kinetic steps of adsorption and desorption of surfactant between an air/water surface and the aqueous bulk sublayer adjacent to the surface. Kinetic constants are determined in nonequilibrium experiments in which either a clean surface is contacted with a bulk solution and surfactant diffuses toward and adsorbs onto the interface, or the area of an established monolayer in equilibrium with an underlying solution is changed, and surfactant exchanges between the surface and bulk. The dynamic tension change due to the surfactant exchange is measured, and compared to predictions of kinetic-diffusive transport models in order to infer the kinetic coefficients as well the diffusion coefficients. Model comparisons for highly surface active surfactants have resolved only the diffusion coefficient as the transport was found to be diffusion controlled; kinetic constants have only been established for less active materials such as alcohols or bolaform surfactants. In this study, we demonstrate that kinetics can be differentiated from diffusion in clean interface adsorption and re-equilibration if high bulk concentrations of the surfactant are used, or in re-equilibration, if the surface is compressed sufficiently. We first establish theoretically that mass transfer shifts from diffusion-limited to mixed as the bulk concentration increases in clean interface adsorption, or the surface compression is increased in re-equilibration. We then experimentally verify this idea by using the polyethoxylated surfactant C12E6 (C12H25 (OCH2CH2)6-OH) and by measuring dynamic surface tensions in clean interface adsorption and re-equilibration, respectively by the shape analysis of pendant bubbles. We find values of 6 x 10(-10) m2/s for the diffusion coefficient, and 1.4 x 10(-5) m/sec and 1.4 x 10(-4) s-1 for the adsorption and desorption rate constants, respectively, in a Frumkin kinetic formulation. While the adsorption constant is comparable to previously measured values for the less surface active surfactants, the desorption rate constant is a few orders of magnitude smaller. This indicates that the more surface active materials may have much smaller desorption rate constants than had been previously anticipated based on the studies of the less surface active materials. Copyright 1998 Academic Press. PMID- 9735186 TI - Solid-Liquid Phase Behavior of Binary Mixture of Tetraethylene Glycol Decyl Ether and Water. AB - Solid-liquid phase behavior of binary mixture of nonionic surfactant, tetraethylene glycol decyl ether (C10E4), and water was examined by means of differential scanning calorimetry (DSC), Fourier transform infrared spectroscopy (FT-IR), polarized optical microscopy (POM), and visual observation in the temperature range -40-70 degreesC. The DSC experiments allowed us to determine many phase boundaries among various phases appearing in this mixture system, including mesomorphic phases. The T-X phase diagram for fluid phases of C10E4/H2O mixture constructed from the results of DSC and visual observation was in good agreement with that previously reported by Lang and Morgan for the same mixture system except for the occurrence of a dilute lamellar phase (Lang, J. C. and Morgan, R. D., J. Chem. Phys., 73, 5849 (1980). The FT-IR spectra obtained for the mixture in a solid state demonstrated that the phase compound expressed by C10E4 . 6H2O is formed in the solid phase, which indicates that there is a strong interaction in the solid state between water and the hydrophilic polyoxyethylene chain of the surfactant. The FT-IR results also suggested that the strength of the hydrogen bond between water and the polyoxyethylene chain of C10E4 differs among the phase states of the mixture, and increases in the order L2 < L1 < Lalpha approximately H1 < solid, where L2 refers to the inverted micellar phase, L1 the normal micellar phase, Lalpha the lamellar phase, and H1 the normal hexagonal phase. Copyright 1998 Academic Press. PMID- 9735187 TI - Effect of Counterion Structure on Micellar Growth of Alkylpyridinium Surfactants in Aqueous Solution. AB - This paper describes the influence of counterions on the unidirectional growth of micelles formed by alkylpyridinium surfactants in aqueous solution. It is shown that the growth of spherical micelles to form wormlike micelles is strongly dependent on counterion structure. More hydrophobic counterions induce the formation of wormlike micelles at lower surfactant concentrations. Next to hydrophobicity and the type of substituent, the substitution pattern of the aromatic ring plays the most important role in micellar growth. The formation of a network of entangled, elongated wormlike micelles by alkylpyridinium surfactants with o-hydroxybenzoate and p-chlorobenzoate counterions is discussed in terms of surfactant structure. It is concluded that, next to counterion structure, the microenvironment of the counterion (substituent) in the Stern region and the structure of the surfactant monomer (i.e., the surfactant cation) play the most important role in the formation of these elongated wormlike micelles. Headgroup effects are proposed to be the main driving force for this phenomenon. Copyright 1998 Academic Press. PMID- 9735188 TI - Binding of Polycarboxylic Acids to Cationic Mixed Micelles: Effects of Polymer Counterion Binding and Polyion Charge Distribution. AB - Mixed micelles of cetyltrimethylammonium chloride (CTAC) and n-dodecyl hexaoxyethylene glycol monoether (C12E8) bind to polyanions when the mole fraction of the cationic surfactant exceeds a critical value (Yc). Yc corresponds to a critical micelle surface charge density at which polyelectrolyte will bind to this colloidal particle. Turbidimetric titrations were used to determine Yc for such cationic-nonionic micelles in the presence of acrylic acid and acrylamido-2-methylpropane sulfonate homopolymers (PAA and PAMPS, respectively) and their copolymers with acrylamide, as function of pH, ionic strength, and polyelectrolyte counterion. In 0.20 M NaCl, Yc for PAA is found to be remarkably insensitive to pH, i.e., virtually independent of the apparent polymer charge density xiapp. On the other hand, the expected inverse relationship between Yc and xiapp is observed either for PAA when NaCl is replaced by TMACl (tetramethylammonium chloride), or when xiapp is manipulated using acrylic acid/acrylamide copolymers at high pH. The effective charge density of PAA is thus seen to be suppressed by specific sodium ion binding, indicating that the influence of salts on the interaction of polycarboxylic acids with colloidal particles may differ qualitatively from their effect on the analogous behavior of strong polyanions. Comparisons between homo- and copolymers of acrylic acid were carried out also to test the hypothesis that the "mobility" of charges on PAA at moderate pH (degree of ionization less than unity) could make this "annealed" polymer exhibit the behavior of a more highly charged one. The results, while consistent with this expectation, were obscured by the likely effect of copolymer sequence distributions. Copyright 1998 Academic Press. PMID- 9735189 TI - Ozone Treatment of Coal- and Coffee Grounds-Based Active Carbons: Water Vapor Adsorption and Surface Fractal Micropores. AB - Characteristics of the adsorption isotherms of water vapor on active carbons from coal and coffee grounds and those ozonized ones from the surface fractal dimension analysis are discussed. The upswing of the adsorption isotherms in the low relative pressure of coffee grounds-based active carbon, of which isotherms were not scarcely affected on ozonization, was attributed to the adsorption of water molecules on the metallic oxides playing the role of oxygen-surface complexes, which formed the corrugated surfaces on the basal planes of micropore walls with the surface fractal dimension Ds > 2. On the other hand, coal-based active carbon with Ds < 2, which indicated the flat surfaces of micropore walls, showed little effect on the upswing even on ozonization, even though the adsorption amounts of water vapor were increased in the low relative pressure. Copyright 1998 Academic Press. PMID- 9735190 TI - Competitive Effects of Nondisplaceable Organic Compounds on Trichloroethylene Uptake by Activated Carbon. I. Thermodynamic Predictions and Model Sensitivity Analyses. AB - Theoretical analyses were performed to investigate potential mechanisms affecting observed reductions in uptake of trichloroethylene from the aqueous phase by activated carbon loaded with nondisplaceable organic molecules. Isotherm sensitivity analysis and thermodynamically based competitive adsorption model predictions give a clear and consistent mechanistic interpretation. At low loadings of nondisplaceable organics, the most significant effect is to reduce the number of high-energy sites available to subsequently adsorbed TCE. The loss of high-energy sites causes a significant reduction in site-energy heterogeneity and reduces TCE uptake in low-equilibrium concentration regions (parts per billion) of the isotherm. As the loading of nondisplaceable compounds increases, further reductions in TCE uptake occur; however, further changes in the site energy heterogeneity are distributed across a wide spectrum of site energies. This suggests a lowering of the average site energy, a reduction in the total number of sites, or both. In terms of TCE isotherms, this corresponds to a roughly constant percentage reduction in uptake over a wide range of equilibrium concentrations, displacing the isotherm downward relative to the uptake axis. Copyright 1998 Academic Press. PMID- 9735191 TI - Competitive Effects of Nondisplaceable Organic Compounds on Trichloroethylene Uptake by Activated Carbon. II. Model Verification and Applicability to Natural Organic Matter. AB - An experimental program was carried out to verify theoretical predictions of competitive effects exerted by nondisplaceable organic compounds on the uptake of TCE by activated carbon. Experimental findings were consistent with isotherm sensitivity analyses and thermodynamically based competitive adsorption model predictions. At low loadings of both trichlorobenzene and a natural humic acid, the most significant effect of preloading was to reduce the number of high-energy sites available to TCE. The loss of these sites caused a significant reduction in the site-energy heterogeneity and reduced the extent of adsorption in the low concentration region. At higher levels of preloading, further changes in the site energy heterogeneity were small, and uptake was reduced by a roughly equal percentage across a wide range of equilibrium concentrations, suggesting the possibility of a pore blockage (in the case of humic acid) or pore filling (in the case of TCB) mechanism. Measurements of adsorbent surface area and pore volume confirmed that observed reductions in TCE uptake by preloaded carbon were associated with changes in the physical characteristics of the adsorbent. However, reductions in adsorbent surface area could only account for a significant fraction ofthe observed reduction in TCE uptake when either the amountpreloaded was high or the TCE concentration was high, increasing the ability of TCE to compete for adsorption sites. Copyright 1998 Academic Press. PMID- 9735192 TI - Computer Simulation of Flocculation Processes: The Roles of Chain Conformation and Chain/Colloid Concentration Ratio in the Aggregate Structures. AB - The flocculation of colloidal particles in the presence of adsorbing polymers is a key process in colloid science, as well as in the chemical and biological regulation of aquatic systems. Polymers can influence important physical properties of colloidal aggregates such as their densities and settling velocities, as well as their chemical properties, affecting the probability that two colloidal particles will stick together when they collide. The presence of polymers usually makes more difficult the application of a coagulation theory to colloidal suspensions and the interpretation of experimental observations. Knowledge of floc structures is a key factor in the understanding of flocculation processes, and simulation may provide useful insights required to interpret the results of experimental studies and elaborate new theoretical models. Although modeling leaves much room for more progress, researchers now find it indispensible from a fundamental point of view and for environmental applications. In this paper, we report a computer simulation study of a two- and three- dimensional model for bridging flocculation between large linear polymer chains and comparatively small colloidal particles. The floc structures are investigated as a function of chain/particle concentration ratio, chain conformation, and space dimension. The values of the sticking probabilities are chosen to emphasize colloid-chain interactions compared to colloid-colloid or chain-chain interactions. The results suggest that the floc morphology is strongly dependent on the chain conformation and to a slight extent on the chain/particle concentration ratio. In particular, colloid interactions with linear rods result in a network characterized by fractal dimensions significantly higher than those obtained on the basis of the Cluster-Cluster Aggregation models of colloids only, or by flocculation of colloids with coiled chains. Copyright 1998 Academic Press. PMID- 9735193 TI - Phase Behavior of Aqueous Mixtures of Some Polyethylene Glycol Decyl Ethers Revealed by DSC and FT-IR Measurements. AB - Differential scanning calorimetry (DSC) and Fourier transform infrared spectroscopy (FT-IR) were used to investigate the phase behavior of aqueous binary mixtures of three polyoxyethylene type nonionic surfactants, penta-, hexa , and octaethylene glycol decyl ethers (C10E5, C10E6, and C10E8, respectively) in the temperature range -40-70 degreesC. Many thermotropic transitions among various phases assumed by these mixture systems, including mesomorphic phases, were detected in DSC thermograms, from which the T-X phase diagrams were constructed with the aid of polarized optical microscopic observation to characterize the mesomorphic phases. It was revealed by FT-IR measurements that the phase compounds are formed in a solid phase between these surfactants and water molecules; the compositions of the compounds for the three surfactant species are expressed as C10E5 . 10H2O, C10E6 . 12H2O, and C10E8 . 16H2O, respectively. The stoichiometry of the phase compounds clearly demonstrates that just two water molecules are bound per oxyethylene unit of polyoxyethylene (POE) chain of the surfactants in the solid phase, probably due to the hydrogen bonding. It was also found that the phase state of the mixture is reflected in the wavenumber of the absorption maximum associated with the OH stretching vibration, nuOH, although no significant difference was appreciable in nuOH among mesomorphic phases. The nuOH decreased in the order L (liquid phase) > H1 (normal hexagonal phase) approximately V1 (normal cubic phase) approximately Lalpha (lamellar phase) > solid phase, which indicates that the hydrogen bonding between water and the POE chain of the surfactants becomes stronger in the sequence liquid < mesophase < solid. Copyright 1998 Academic Press. PMID- 9735194 TI - The Effects of Caseinate Submicelles and Lecithin on the Thin Film Drainage and Behavior of Commercial Caseinate. AB - The drainage behaviour (stratification, thickness, and mobility) of thin foam films stabilized by commercial caseinate was studied in 10 mM phosphate buffer at pH 7.0. Thin films of commercial caseinate drained in a stepwise manner, with steps of similar thickness. The drainage was rapid, temperature sensitive, and chaotic, and the surface mobility of caseinate thin films also showed temperature sensitivity. The stepwise drainage is thought to be due to the layering of lecithin-caseinate submicelle complexes. Lecithin-stabilized thin films showed similar drainage behavior and temperature sensitivity. However, the films were approximately 66% thinner than caseinate films, and surface diffusion was very rapid. Removal of lipid from caseinate dramatically affects the thin film drainage properties and reduces temperature sensitivity. Reconstituted caseinate (i.e., extracted caseinate reconstituted with lipid), showed thin film properties similar to the commercial caseinate. Caseinate supplemented with lipid showed thin film drainage characteristics similar to caseinate, and surface mobility similar to lecithin. The presence of lecithin in caseinate thin films causes an increase in mobility, drainage, and stratification, along with a decrease in thin film thickness. This demonstrates that lecithin, possibly partially bound to the caseinate, is present at the interface disrupting protein-protein interactions. Copyright 1998 Academic Press. PMID- 9735195 TI - Interaction of Different Types of Cells on Polymer Surfaces with Wettability Gradient. AB - Gradient surfaces whose properties are changed gradually along the sample length are of particular interest for basic studies of the interaction between biological species and surfaces since the effect of a selected property can be examined in a single experiment on one surface. We prepared a wettability gradient on low density polyethylene (PE) sheets by treating them in air with the corona from a knife-type electrode whose power increases gradually along the sample length. The PE surfaces oxidized gradually with the increasing corona power, and the wettability gradient was created on the surfaces as evidenced by the measurement of water contact angles, Fourier transform infrared spectroscopy in the attenuated total reflectance mode, and electron spectroscopy for chemical analysis. The wettability gradient surfaces prepared were used to investigate the interaction of different types of cells (Chinese hamster ovary, fibroblast, and endothelial cells) as well as serum proteins in terms of the surface hydrophilicity/hydrophobicity of polymeric materials. The cells adhered and grown on the gradient surface along the sample length were counted and observed by scanning electron microscopy. It was observed that the cells were adhered, spread, and grown more onto the positions with moderate hydrophilicity of the wettability gradient surface than onto the more hydrophobic or hydrophilic positions. The maximum adhesion and growth of the cells appeared at around water contact angles of 55 degrees, regardless of the cell types used. This result seems closely related to the serum protein adsorption on the surfaces; the serum proteins were also adsorbed more onto the positions with moderate hydrophilicity of the wettability gradient surface. Copyright 1998 Academic Press. PMID- 9735196 TI - Hydrophobic Aggregation in Polyurethane Ionomer Solutions. AB - Model polyurethane ionomers were synthesized with regularly spaced ionic groups along the polymeric backbone. Viscometry and static and dynamic light scattering applied to these ionomer solutions in a polar solvent, N-methylformamide, revealed hydrophobic interactions and aggregated ionomer chains in solution. A new regime in the solution behavior of ionomers characterized by low reduced viscosity at all dilute concentrations has been identified. Spherical particles of radius 30 nm with a polymer core and an outer ionic shell are formed. The solutions have very low viscosity because of the formation of compact structures. The electrostatic interactions in these solutions are effectively screened, resulting in the absence of polyelectrolyte behavior. Polymer-solvent interactions are found to be very important in determining the ionomer solution behavior in polar solvents. Copyright 1998 Academic Press. PMID- 9735197 TI - Preparation and Properties of Monodisperse Magnetic Cobalt Colloids Grafted with Polyisobutene. AB - A single-step method to synthesize monodisperse metallic cobalt particles of diameter around 8 nm is described. The particles are sterically stabilized by modified polyisobutene and form stable ferrofluids in toluene. The core-shell cobalt spheres have a narrow size distribution and are nonaggregated. Various techniques, such as infrared spectroscopy, X-ray diffraction, and elemental analysis establishes the presence of grafted modified polyisobutene on the surface of cobalt particles. The hydrodynamic thickness of the grafted polyisobutene calculated from sedimentation and viscosity measurements and independently from the dynamic light scattering is ca. 10 nm. The dependence of magnetization has been measured for ferrofluid, showing superparamagnetic behavior above 247 K. Due to anisotropy, superposition of magnetic curves is not observed below this temperature. The concentration dependence of the sedimentation coefficient agrees qualitatively with the theory for dipolar spheres. SAXS data on concentrated ferrofluid dispersion indicate interpenetration of polyisobutene chains grafted to the cobalt particles. Copyright 1998 Academic Press. PMID- 9735198 TI - Electrostatic Interaction and Hindered Diffusion of Ion-Penetrable Spheres in a Slit Pore. AB - Electrostatic interaction and hindered diffusion of ion-penetrable spheres in a slit pore filled with an electrolyte solution are investigated theoretically. The concentration of the particles is assumed sufficiently low so that the interactions between the particles can be neglected. The slit pore comprises two parallel infinite plates, which can be either permeable or impermeable to the electrolyte. The electrostatic interaction energy is obtained analytically by adopting an integral of Green's function and an image method. It is found that for impermeable plates having constant surface charge densities, the electrostatic interaction can be attractive or repulsive for a particle with charge of opposite sign, depending on the relative magnitudes of charge densities and particle location. Similar behavior is predicted for plates with constant surface potential and a particle with charge of like sign. The interaction energy is used to determine the spatial distribution of the particle in the pore and the partition coefficient, and then to calculate the average and apparent diffusivities. The average diffusivity calculated from the average mobility in the pore is always smaller than that in the bulk solution because of greater drag the particle experiences in the presence of the plates. This mean diffusivity is larger than that in the corresponding neutral system for repulsive electrostatic interaction, but becomes smaller for attractive interaction. The apparent diffusivity defined from the flux based on the bulk concentration of the particle depends strongly on the ion concentration, as does the partition coefficient when the double layer is sufficiently thick. Copyright 1998 Academic Press. PMID- 9735199 TI - Current-Voltage Curves for Ion-Exchange Membranes: A Method for Determining the Limiting Current Density. AB - The curves (V, I) of different ion-exchange membranes have been determined under different experimental conditions. Taking into account the classical theory of concentration polarization, a method has been developed to adjust the (V, I) data, up to a certain value of I, which permits us to obtain easily the value of the limiting current density, IL, in every experimental situation. From these values, the dependence of the limiting current density of the membranes used on the concentration and stirring rate of the solutions has been studied. To check the reliability of the method, the values of IL thus obtained have been compared with those obtained by the usual methods; the similitude between both values is high. Copyright 1998 Academic Press. PMID- 9735200 TI - Separation of the Intramembrane Diffusion Potential and the Donnan Potential on the Basis of the Potential Transient Measurement: Application to the Analysis of Reverse Ion Permeation Driven by pH Difference. AB - In the perfluorocarboxylate ion exchange membrane-aqueous sodium chloride system, the diffusional flux of sodium ions against their own concentration difference was observed in the presence of a pH difference across the membrane. The internal solution contained 1 x 10(-1) mol dm-3 NaOH and 1 x 10(-1) mol dm-3 NaCl, and the external solution contained 2 x 10(-1) mol dm-3 NaCl and HCl of various concentrations in the range of 1 x 10(-2) to 1 x 10(-1) mol dm-3. In these membrane systems, it was observed that the membrane potential rapidly changed in response to a pH jump in the external side of two aqueous phases to reach an intermediate stage and then the subsequent step started to relax slowly to the final membrane potential at the other steady state. On the basis of the assignment that the earlier fast step and subsequent slow step observed in the generation process of the membrane potential are the generation processes of the Donnan potential at the membrane/solution interface and the intramembrane diffusion potential, respectively, the total membrane potential has been divided into these two constituents. By using the observed Donnan potential, the ion concentration at the membrane surface in the membrane was obtained. The ion flux was analyzed to obtain the diffusion coefficient of ions within the membrane by using the ion concentration at the membrane surface in the membrane and the intramembrane diffusion potential. The pH dependence of the diffusion coefficient and that of the ion concentration at the surface in the membrane showed a break point near the apparent pKa, where the transition of the ion cluster structure in the membrane would occur. Copyright 1998 Academic Press. PMID- 9735201 TI - Binding Characteristics of Pb2+ on Anion-Modified and Pristine Hydrous Oxide Surfaces Studied by Electrophoretic Mobility and X-Ray Absorption Spectroscopy. AB - We examine the role of coadsorbed phosphate and sulfate on the adsorption of Pb2+ at the oxide-water interface of goethite and boehmite by electrophoretic mobility (EM) and X-ray absorption spectroscopy (XAS). Comparisons to Pb2+ adsorbed to pristine oxides are provided. Pb2+ binds as an inner-sphere complex to pristine goethite and groups into oxy-clusters on pristine boehmite under our conditions. The presence of either anion at the surface generally increases the ability of the solids to sorb Pb2+ but by different mechanisms. Adsorbed phosphate on both solids acts as a reactant to form lead phosphate surface phases that may be highly dispersed. The more mobile sulfate ion is more effectively adsorbed when Pb2+ is present, and increases Pb2+ adsorption on boehmite. Lead sulfate phases may form at the goethite surface under our conditions. Copyright 1998 Academic Press. PMID- 9735202 TI - Influence of Polarity and Viscosity of the Micellar Interface on the Fluorescence Quenching of Pyrenic Compounds by Indole Derivatives in AOT Reverse Micelles Solutions. AB - The fluorescence quenching of the pyrene derivatives (4-(1-pyrenyl)butyl) trimethylammonium bromide (PBTMA), (1-(1-pyrenyl)methyl) trimethylammonium iodide (PMTMA), and 1-pyrene sulfonic acid (PSA) by indole methyl substituted in positions 1 and 2, tryptophan and tryptamine, was studied in AOT/heptane reverse micelles as a function of R = [water]/[AOT]. In these systems the pyrenic probes are associated to the micellar interface. Bulk and intramicellar quenching rate constants were determined for neutral indoles. The quenching rate constants of PBTMA by indole or 1,2-dimethylindole increase with R, whereas for those for PMTMA or PSA by indole the increment is much smaller. For the quenchers, tryptophan and tryptamine, that are bound to the interface of the reverse micelle, the bimolecular intramicellar quenching rate constant is much lower than in water. The results can be explained by a high microviscosity of the interface, and a micropolarity similar to that sensed by other probes. Moreover, the observed trend in the rate constants when R is varied is in line with the reported changes in micropolarity and microviscosity. Laser flash photolysis experiments show that in these systems the main result of the quenching process is the formation of the excited triplet of the probe. Copyright 1998 Academic Press. PMID- 9735203 TI - Rheo-optical Behaviors and Stability of a Silica Particle Suspension Coated with Silane Coupling Agents. AB - The rheo-optical behaviors and suspension stability of silica particles coated with silane coupling agents were investigated experimentally. Mono-dispersed silica particles were synthesized by the sol-gel method and the particles were coated with silane coupling agents such as vinyltriethoxy silane (VTES) and gamma methacryloxypropyl triethoxy silane (MPTES). Although all the suspensions of identical particle volume fraction exhibited similar rheological behaviors at high shear rates, only the stabilized suspensions coated with either VTES or MPTES displayed smooth shear thinning rather than abrupt change in the shear viscosity, as is typical of suspensions with no surface treatment. The present study showed that the MPTES coating was very effective in enhancing the phase stability compared to the VTES coating. The flow-induced dichroism for the MPTES coated suspensions did not experience sign change, while those of the non stabilized suspensions changed sign as the shear rate was increased. The VTES coated suspensions underwent a transition from a stable to an unstable state as the particle volume fraction increased. Specifically, the rheological behaviors of the VTES-coated suspensions of particle volume fraction 0.35 were similar to those of the MPTES-coated suspensions. When the volume fraction exceeded 0.45, however, the effect of VTES coating diminished. Finally, the stress-optical rule proposed by Bender and Wagner was adopted to model the stabilized suspensions considered here. The present results indicated that the stress-optical coefficient could be predicted successfully by the proposed stress-optical rule if the contribution from the hydrodynamic interaction is considered separately from the thermodynamic contribution. Copyright 1998 Academic Press. PMID- 9735204 TI - Structural Environment of Uranium (VI) and Europium (III) Species Sorbed onto Phosphate Surfaces: XPS and Optical Spectroscopy Studies. AB - In order to characterize the structure of the surface complexes formed by interaction between uranyl and europium (III) ions and the surface of solid matrices, optical and X-ray photoelectron spectroscopies experiments on uranyl/europium loaded phosphate solids (Th4P2O7(PO4)4, ZrP2O7, and Zr2O(PO4)2) have been performed. The use of complimentary spectroscopic techniques allows an identification of the sorption mechanism and a structural characterization of the sorption sites and the sorbed species on phosphate surfaces. The samples were prepared from aqueous uranyl or europium solutions in the pH range from 1.5 to 6.0. The surface coverage was varied from 1 to 40 % of a monolayer. The differences between the emission spectra of europium ions either sorbed on the surface of phosphate samples or doped inside the solid unambiguously indicates that these sorbed ions are exclusively located on the surface and that they do not migrate inside the matrix, which shows clearly that surface complexation is involved during the sorption process. The U4f XPS spectrum of uranyl ions sorbed on zirconium diphosphate exhibits only one component, while the spectrum corresponding to uranium on thorium matrix shows two different unresolved peaks attributed to two different chemical environments. These results, corroborated by the uranyl emission spectra and the associated decay times and those obtained by optical spectroscopy of europium sorbed on the same solids, have been interpreted in terms of two sorption sites probably formed by the oxygens of the PO4 and P2O7 surface groups. Copyright 1998 Academic Press. PMID- 9735205 TI - Surface Properties of Fluorosilicone Copolymers and Their Surface Modification Effects on PVC Film. AB - The fluorosilicone copolymers were synthesized using a fluorine-containing monomer and silicone-containing monomers by free-radical random copolymerization, and their surface properties and surface modification ability were investigated. The fluorine-containing monomer used was perfluoroalkyl ethyl acrylate (FA), and the silicone-containing monomers used were 3-[tris(trimethylsilyloxy)silyl]propyl methacrylate (SiMA), vinyltrimethoxy silane (VTMS), and vinyltriethoxy silane (VTES). The surface free energies of the fluorosilicone copolymers prepared were estimated from the contact angle data measured by sessile-drop method. And, the surface free energies of poly(vinyl chloride) (PVC) films modified by the fluorosilicone copolymers were also analyzed using the contact angle data. The fluorosilicone copolymers exhibit the surface free energies of about 8-23 dyn/cm dependent on the molecular weight of the fluorosilicone copolymers. The surface free energies of the fluorosilicone copolymers decrease with increasing molecular weight in the range of 2,000-10,000 (Mw). Among the fluorosilicone copolymers prepared in this study, PFA-r-PSiMA was found to be the most effective as a surface modification agent for PVC film. The inherent surface free energy of PFA r-PSiMA was estimated to be about 9.0 dyn/cm. The desirable molecular weight of PFA-r-PSiMA seems to be more than 4,000 (Mw). However, it is expected that the fluorosilicone copolymers having the molecular weight of much higher than 10,000 (Mw) may not be suitable as surface modification additives because their compatibility with other polymers will decrease with the molecular weight. The optimum concentration of PFA-r-PSiMA added to PVC film is about 1.0 wt.%. PFA-r PSiMA is expectedto be an effective additive for surface modification of PVC films. Copyright 1998 Academic Press. PMID- 9735206 TI - Electrical Double-Layer Interactions of Regular Arrays of Spheres. AB - Electrostatic interactions are computed for three-dimensional regular arrays of spheres. Tetrahedral array of four spheres, octahedral array of six spheres, cubic array of eight spheres, and periodic arrays of spheres in body-centered cubic (BCC) and face-centered cubic (FCC) structures are considered. Based on the linearized Poisson-Boltzmann equation the electrostatic free energy is computed for both constant surface potential and constant surface charge conditions. Facilitating the complex geometry of the problem, the boundary element method is used. Test results for two-sphere interactions are in excellent agreement with existing exact solutions even at very small Debye length. The results for multisphere interactions are compared with those obtained by the pairwise additivity approximation. For BCC and FCC arrays comparison is made to the results from the pairwise additivity approximation and from the spherical cell model. At small Debye length the pairwise additivity approximation predicts the interactions reasonably well. At large Debye length, however, the cell model furnishes much better estimates. In addition, the electrostatic free energy difference between BCC and FCC structures at given particle concentration is found to be too small to assess any physical significance. Copyright 1998 Academic Press. PMID- 9735207 TI - Surface Tension of Biological Polyelectrolyte Solutions. AB - Surface tensions, gamma, of biological polyelectrolytes in aqueous solutions are studied systematically as possible at the air-water interface by the Wilhelmy method. The polyelectrolytes measured are sodium chondroitin sulfates A (NaCRA) and C (NaCRC), sodium poly-alpha,l-glutamate (NaPGA), poly-l-lysine hydrobromide (PLL . HBr), deoxyribonucleic acid (DNA), lysozyme (LZ), and bovine serum albumin (BSA). Linear-type macroions such as NaCR, NaPGA, PLL . HBr, and DNA have no surface activity in a wide range of polymer concentrations below the critical polymer concentration, m*, and increases as the concentration increases above m*. Surface activity of the undissociated state of macroions is rather high in general. Globule-like macroions such as LZ and BSA show high surface activity at isoelectric point above m* accompanied with orientation of the molecules along the air-water interface. Separation into the hydrophobic and hydrophilic parts at the interface and balancing in their strength are important for appearance of surface activity. Copyright 1998 Academic Press. PMID- 9735208 TI - Intramolecular Excited State Proton Transfer of 2-(2'-Hydroxyphenyl)benzimidazole in Nonionic Micelles: Tweens. AB - Spectral characteristics and prototropic reactions of 2-(2' hydroxyphenyl)benzimidazole (HPBI) in Tween-20, Tween-40, Tween-60, Tween-80, and dioxane-water mixtures of different compositions have been studied. Comparison of the fluorescence band maxima of the tautomer band in nonionic micelles with the correlation diagram, drawn between the fluorescence band maxima and the dielectric constants of the dioxane-water mixtures, have shown that the effective dielectric constant (epsiloneff) at the binding site of HPBI is 18 +/- 2 for all the Tweens. Fluorescence lifetimes (tauf) and fluorescence quantum yields (phifl) have shown that the hydrophobicity of these micelles is maximum for Tween-80 and at a minimum for Tween-20. Similar results have also been observed from the pKa values for the deprotonation of HPBI, whereas the protonation reaction of HPBI occurs at the site in nonionic micelles which is more hydrophilic than that where the deprotonation reaction takes place. The protonation reaction in 2-(2' methoxyphenyl)benzimidazole (MPBI) has shown that the value of epsiloneff in Tweens where this reaction occurs is less hydrophobic than the site where the same reaction occurs in HPBI. Copyright 1998 Academic Press. PMID- 9735209 TI - The Interaction of Micelles with Added Species and Its Similarity to the Denaturant Binding Model of Proteins. AB - The effect of additives on micellar aggregation is treated using the same framework developed for macromolecular binding. This procedure, hinging on the pseudo-phase transition model, allows us to evaluate the binding constant of any added substance to a micellar aggregate as well as the number of particles bound per amphiphilic unit. The analysis of published literature data confirms that the model provides a general description of micellar aggregation in the presence of added ingredients, regardless of their ionic or non-ionic nature. Original data from our laboratory are well described by the suggested macromolecular binding approach. The dependence of experimental trends on micellization modifiers is also briefly discussed. Copyright 1998 Academic Press. PMID- 9735210 TI - Fractal Aggregates of Polydisperse Particles. AB - In this work we examine structural effects of particle polydispersity on fractal aggregates by performing DLCA simulations with multiple primary particle sizes. We show that the fractal structure and the form of the cutoff function that describes the gross shape of the aggregates is unaffected by the details of the primary particle size distribution. The scattering behavior is evaluated in terms of partial structure factors, and depending on the details of primary particle size distribution and contrast, the scattering curve can deviate significantly from the typical q-Df behavior at the high q end of the spectrum. We also develop an expression for average primary particle size that allows the calculation of aggregate solid volume fraction for fractal aggregates with polydisperse primary particles. Copyright 1998 Academic Press. PMID- 9735211 TI - Interfacial and Rheological Characteristics of Maghemite Aqueous Suspensions. AB - The interfacial properties and rheological behavior of the maghemite/water system have been investigated in this work. It has been determined that the real pzc and iep values of maghemite immersed in aqueous solution, containing KNO3 as electrolyte, are very close to 6.6. It has also been shown that the triple-layer site-binding model likely describes the electrical double layer of the maghemite/water system. The parameters obtained from this model indicate that the mechanism responsible for charging involves complexation reactions and that the interfacial properties of aqueous suspensions containing maghemite are very similar to those containing magnetite. Finally, rheological characterization of three maghemite samples with different average particle size showed that the maximum yield stress, at a fixed solid concentration, decreases as the sample particle size decreases. This anomalous behavior is originated by the apparition of relaxation superparamagnetic phenomena as a consequence of the decrease in the sample particle size. Copyright 1998 Academic Press. PMID- 9735212 TI - Nucleation and the Tunneling Phenomenon. AB - The rate of nucleation of a stable phase in a homogeneous metastable medium is computed within the context of particle tunneling through a modified cubic potential, bypassing the conventional thermodynamic approach. Comparison with the standard Becker-Doring results hints at good prospects for the present procedure. Some fundamental issues are raised regarding current approaches. Copyright 1998 Academic Press. PMID- 9735213 TI - Interfacial Free Energies and Surface Areas of Cycloalkanols and alpha,omega Alkanediols at the Air-Water Interface. AB - The surface area occupied at the air/water interface for cycloalkanols and alpha,omega-alkanediols has been estimated from surface tension measurements. From these data, standard free energies of transfer of the alkanols from the bulk of the solution to the interface were obtained. Cycloalkanols constitute a family whose surface excess diminishes when the size of the alkyl ring increases and whose free energies of transfer at zero surface pressure are similar to those of 1-alkanols of similar hydrophobicities. alpha,omega-Alkanediols occupy considerably larger surface areas than 1-alkanols, a result that can be interpreted in terms of a different geometrical arrangement of the molecules at the interface. In particular, for alkanediols with n > 7, the data suggest an important contribution of bent conformations to the average arrangement of the molecules in the monolayer. The need to adopt bent conformations to allow the interaction of both hydroxyl groups with the aqueous interface results in a less favorable change in free energy when they are transferred from the solution standard state to the interface at constant surface pressure. Copyright 1998 Academic Press. PMID- 9735214 TI - Adsorption and Reaction Behavior of Perfluoro-n-alkanes on Al2O3. AB - The adsorption of perfluoro-n-pentane and perfluoro-n-hexane on gamma-Al2O3 at the solid/vapor interface has been studied by infrared spectroscopy and temperature-programed desorption (TPD). At ambient beam temperature perfluorocarbons are adsorbed as intact, unchanged molecules and undergo chemical transformations leading to the formation of adsorbed, unsaturated carboxylate species. With increasing temperature further reactions occur, finally resulting in deep oxidation of all adsorbed species (indicated by desorption of CO and CO2). These reactions are initiated by the formation of coordination bonds between perfluorocarbons and Lewis-acidic surface sites (coordinatively unsaturated cations) of Al2O3, followed by abstraction of fluorine from the adsorbed molecules. Copyright 1998 Academic Press. PMID- 9735215 TI - Comparing the Surface Chemical Properties and the Effect of Salts on the Cloud Point of a Conventional Nonionic Surfactant, Octoxynol 9 (Triton X-100), and of Its Oligomer, Tyloxapol (Triton WR-1339). AB - The surface-chemical properties, critical micelle concentrations (CMC), and effect of salts on the cloud points (CP) of octoxynol 9 (Triton X-100) and tyloxapol (Triton WR-1339) were compared. The latter nonionic surfactant is essentially a heptamer of the former. Even though the molecular weight of tyloxapol is 7 times larger than that of octoxynol 9, its area per molecule adsorbed at the air-water interface is only twice as large. This suggests an unusual orientation for molecules of tyloxapol at the surface and is in keeping with a plateau that is less horizontal and has a somewhat higher surface tension than the plateaus of most nonionic surfactants. The CMC of octoxynol 9 was 4.4 times larger than that of tyloxapol. Unexpectedly, the CP of dilute aqueous tyloxapol solutions was 28 degreesC higher than that of octoxynol 9 solutions. The salting-out ions Na+, Cl- and SO2-4 lowered the CP of tyloxapol 29% more than that of octoxynol 9. However, because the blank tyloxapol solution started out with a higher CP value, its CPs in the presence of salts were higher than those of octoxynol 9. Pb2+ and Mg2+ cations salted both surfactants in, raising their CP, Pb2+ more extensively than Mg2+. Copyright 1998 Academic Press. PMID- 9735216 TI - Interphase Transport of Benzoic Acid in Emulsions. AB - The kinetics of transport of benzoic acid in triglyceride-in-water emulsions stabilized by Tween 80 was measured by a continuous flow kinetic apparatus. The determination of the liquid-liquid partitioning of the solute in two-phase systems enabled estimation of the retention of benzoic acid by the triglycerides and the micellar solution and calculation of the solute theoretical distribution in emulsions. The initial rates of release of benzoic acid from 2 to 5 mmol m-2 s 1 were found proportional to the initial concentration of benzoic acid in the emulsions. Copyright 1998 Academic Press. PMID- 9735218 TI - Perikinetic Aggregation Induced by Chromium Hydrolytic Polymer and Sol. AB - We investigated the kinetics of destabilization of polystyrene latex particles bearing sulfate surface groups in the presence of aged chromium polymer and oxide sol particles of relatively smaller size employing a particle counting technique. From the mass distribution curve, we derived the weight S(t) and number N(t) average masses of the aggregates. From the variation with time of these characteristics we determined the scaling exponents z and w of the kinetic laws which serve to characterize the aggregation mechanism induced by the inorganic polymer and sol. From the slope of the self-preserved reduced mass distribution, we determined the exponent tau, and found the relationship tau = 2 - (w/z) to be valid. The usual value of tau = 1.5 was obtained for experiments performed in the presence of very small amount of polymer. Deviation from this behavior was determined in the presence of increasing amounts of polymer and sol. At short and long terms, we determined tau to be equal to 1.24 and 1.37, respectively. This was attributed to the diffusion-limited aggregation accompanying the prevailing reaction-limited process, which was found to provide an unusual mass distribution frequency characterized by a hump in the monotonically decreasing curve. From the comparison between the aggregation rates and the apparent aggregate reactivity at short and long terms, we concluded that (i) the short-term aggregation involving aggregates of high apparent reactivity is highly reversible and (ii) the long term aggregation involving aggregates of lower apparent reactivity is irreversible (or less reversible). Copyright 1998 Academic Press. PMID- 9735217 TI - Fluorometric Studies of Pyrene Adsorption on Porous Crystalline Cellulose. AB - Pyrene crystals were physically mixed with either porous crystalline cellulose (PCC) or octa decyl siryl silica-80Tm (ODS). Solid-state fluorescence spectra of pyrene were analyzed to estimate the interaction between pyrene and porous materials. Pyrene monomer emission was observed at 398 nm immediately after being mixed with PCC, while pyrene crystals showed only excimer emission at 475 nm, indicating that the pyrene molecules adsorbed onto the PCC surface in a short period. For the PCC system containing 1.0% pyrene, long-term storage caused an increase in the intensity of excimer-like emission peak at 477 nm accompanied by a decrease in the intensity of monomer emission peak at 398 nm. For the ODS system containing 1.0% pyrene, spectrometric changes were similar to those for the PCC system. In the process of interaction formation between pyrene and an additive, a two-step mechanism was proposed, i.e., the adsorption of pyrene molecules onto the surface of porous additives, and the formation of a ground state dimer of pyrene. The formation of dimeric pyrene could be associated with the surface polarity of additives. Copyright 1998 Academic Press. PMID- 9735219 TI - Adsorption Characterization of Two Clay Minerals Society Standard Kaolinites. AB - The kaolinite samples KGa-1b and KGa-2 are used as standards by the Clay Minerals Society of America. Recently a comparative study of these samples was carried out using atomic force microscopy (AFM) and X-ray diffraction methods (XRD). The aim of the current work was to characterize the kaolinite standards by high resolution nitrogen adsorption and to investigate how their surface heterogeneity changes upon sample cleaning. Copyright 1998 Academic Press. PMID- 9735220 TI - The Effect of Surfactant Monolayers on the Heat Transfer Through Air/Water and Oil/Water Interfaces Using IR Imaging Technique. AB - An experimental investigation on the effect of surfactant monolayers on the heat transfer through air/water and oil/water interfaces was carried out by observing the changes of surface temperature with IR Imaging Radiometer (Model 760). The heat transfer resistance of various single component and mixed monolayers at air/water and oil/water systems was studied. The results show that the surfactant monolayers introduce a noticeable heat transfer resistance to the heat transfer process across the interface. The solid monolayers exhibit lower resistance to heat transfer than the liquid monolayers at the oil/water interface. At air/water interface, the presence of monolayer decreases the evaporative cooling process and therefore increases the surface temperature rapidly. However, the presence of a monolayer at oil/water interface increases the heat transfer resistance across the oil/water interface. Heat transfer resistance increases as the chain length of fatty acid increases at the oil/water interface. The effects of phase transition from a two-dimensional solid to the liquid state in cholesterol arachidyl mixed monolayers was observed from the change in heat transfer resistance of the monolayers at the oil/water interface. The optimum molecular packing at the 1:3 molecular ratio in mixed surfactant monolayers of oleic acid cholesterol and stearic acid-stearyl alcohol at the oil/water interface was also observed by this technique. Copyright 1998 Academic Press. PMID- 9735221 TI - Synthesis of Ion Conducting Polymer Protected Nanometer Size Platinum Colloids. AB - Pt particles have been prepared by reduction of hexachloroplatinic acid with ethylene glycol in the presence of a negatively charged polymer poly(n sulfonatopropyl p-benzamide). The average size of platinum particles was investigated by transmission electron microscopy and found to be in the nanometer size range. The effects of the polymer content on the average particle size were studied. The average particle size of the Pt colloid prepared from solutions containing a polymer/Pt weight ratio of 20 and 0.2, respectively, varied from 2.45 to 5.9 nm. Copyright 1998 Academic Press. PMID- 9735222 TI - Electrostatic Interactions between Two Plates Covered with Charge-Regulation Layers. AB - The interaction energy between two plates covered with charge-regulation layers is calculated based on a linearized Poisson-Boltzmann equation. By a comparison with the exact solution, the accuracy of the approximate solutions obtained by linear superposition approximation and the regular perturbation method can be examined. The range of the validity for both approximations is therefore determined. These results may shed some light on the interaction between two spheres, which is complicated and often investigated by the approximate approaches. Copyright 1998 Academic Press. PMID- 9735224 TI - Effect of long-term castration and long-term androgen treatment on sexually dimorphic estrogen-inducible progesterone receptor mRNA levels in the ventromedial hypothalamus of whiptail lizards. AB - In whiptail lizards, as in laboratory rodents, females will respond to exogenous estrogen by increasing progesterone receptor (PR) or PR mRNA in the ventromedial hypothalamus (VMH) while males show an attenuated response to the same treatment. In rodents, neonatal hormone manipulations affect the adult expression of this trait; however, few investigators have examined the effects of hormone treatment in adulthood. Therefore the current study was carried out to determine whether observed sex differences in the estrogen response in adulthood may be modified by steroid hormone manipulation. We castrated male whiptail lizards for 1 week (short term) or 6 weeks (long term). We also gonadectomized female whiptails and implanted them with either a Silastic capsule containing testosterone or an empty capsule. At the end of that time all implants were removed and the animals were injected with either estradiol benzoate (EB) or steroid suspension vehicle and their brains were assayed for PR mRNA expression using in situ hybridization. The results demonstrate that in male whiptail lizards, long-term castration increases sensitivity to estradiol as measured by induction of PR mRNA in the VMH; EB injected long-term castrated males were not different from EB-injected females. However, long-term androgenization did not attenuate the estrogen response in females. This suggests that attenuation of the estrogen response in males requires activation by testicular secretions, but that females cannot be made to show a male phenotype via testosterone administration. PMID- 9735223 TI - The effect of exogenous testosterone on parental behavior, plasma prolactin, and prolactin binding sites in dark-eyed juncos. AB - Numerous studies have shown that parental behaviors are mediated by prolactin (PRL), while testosterone (T) interferes with their full expression. The limited data available suggest that reduced parental behavior induced by T is not mediated by reduced concentrations of plasma PRL. We hypothesized that T reduces parental behaviors by reducing PRL receptor binding activity at central neural sites that promote the expression of parental behaviors. To test this hypothesis we implanted male dark-eyed juncos (Junco hyemalis) with testosterone-filled or empty implants and measured T and PRL levels, paternal behavior, and specific binding of radio-labeled PRL at selected brain regions that have been implicated in the mediation of parental behaviors. Our findings concurred with previous studies in that T-treated males reduced their parental contributions, had higher levels of T, and had equivalent levels of PRL compared with controls. We found no differences in the capacity to bind 125I-oPRL in three brain regions previously implicated in the mediation of parental care in birds, i.e., the preoptic area, ventromedial nucleus of the hypothalamus, and paraventricular nucleus of the hypothalamus. Thus our findings do not support the hypothesis that T interferes with the expression of parental behavior by reducing PRL receptor binding activity at central sites. PMID- 9735225 TI - Fos-immunoreactivity within the extended amygdala is correlated with the onset of sexual satiety. AB - We hypothesized that c-fos expression in the medial amygdala (Me), the bed nucleus of the stria terminalis (BNST), and the medial preoptic area (MPOA) of the male Syrian hamster brain correlated with sexual satiety. To address this hypothesis, males were mated for 4 consecutive days. Experiment 1 determined whether the number of Fos-immunoreactive (Fos-ir) nuclei was equivalent in two groups of males mated to sexual satiety, one group of rested males (9.67 +/- 0.80 ejaculations) and a second group mated for 4 consecutive days (3.50 +/- 0.56 ejaculations). Fos-ir was increased within the caudal posterodorsal Me (cMePD), the anterodorsal and posteroventral subdivisions of the posteromedial BNST [BNSTpm(ad) and BNSTpm(pv)], the dorsolateral MPOA, and the medial preoptic nucleus of all males mated to sexual satiety compared to nonmated controls. In addition, Fos-ir "clusters" within the cMePD and BNSTpm(ad) were present in males mated to satiety regardless of the number of ejaculations. However, all males achieved multiple ejaculations. Therefore, Experiment 2 examined whether two groups of males stopped at one ejaculation exhibit different patterns of Fos-ir depending on proximity to sexual satiety. Brains of consecutively mated males, closer to satiety than rested males, showed greater BNSTpm(pv) Fos-ir and 5/6 males, but no rested male, exhibited cMePD Fos-ir clusters. These results support the hypothesis that cMePD and BNSTpm(pv) neuronal activation is associated with satiety and may constitute a discrete circuit to terminate mating. PMID- 9735226 TI - Behavioral actions of androgens and androgen receptor expression in the electrocommunication system of an electric fish, Eigenmannia virescens. AB - Gymnotiform electric fish emit an electric organ discharge (EOD) that, in many species, is sexually dimorphic and used in gender recognition. The glass knife fish Eigenmannia has been a major neuroethological model system, and there have been reports that its EOD is sexually dimorphic. However, no study has examined the role of steroids in establishing this dimorphism. We tested the effect of three androgens, testosterone, dihydrotestosterone, and 11-ketotestosterone (11kT), on the EOD of Eigenmannia virescens. Implants of any of these three androgens induced a decrease in EOD frequency, consistent with several studies showing that males have a lower EOD frequency than females. In a separate experiment, we found that 11kT treatment also increases EOD pulse duration, but has no effect on the duty cycle, the relative duration of the positive and negative phases of the wave, or the harmonic content of the EOD. Using immunocytochemistry, we found that the cells of the electric organ (the electrocytes) that produce the EOD and control its pulse duration label positively with an antibody to the androgen receptor. These experiments indicate that androgens decrease the firing frequency of the medullary pacemaker and alter the ion current kinetics in the electrocytes. Moreover, the presence of nuclear androgen receptors in electrocytes suggests that androgens act directly on the electric organ to modify EOD pulse duration. PMID- 9735228 TI - Pheromones elicit equivalent levels of Fos-immunoreactivity in prepubertal and adult male Syrian hamsters. AB - Male reproductive behavior in the Syrian hamster is dependent on both pheromones from the female and the presence of gonadal steroid hormones. The pheromones are contained within female hamster vaginal secretions (FHVS) and stimulate anogenital investigation and mounting by the male. Administration of testosterone to castrated male hamsters facilitates anogenital investigation, mounts, and intromissions in adults, but elicits only anogenital investigation in prepubertal males. One hypothesis for why the full complement of reproductive behaviors is not activated by testosterone in prepubertal males is that the neural processing of pheromonal cues encountered during anogenital investigation is different in juveniles and adults. In the present experiment, we investigated the influence of sexual maturity on Fos expression in response to FHVS in the male Syrian hamster. We predicted a greater increase in Fos-immunoreactivity after exposure to FHVS within the neural circuit mediating male reproductive behaviors in adult compared to prepubertal males. Intact adult and prepubertal males were exposed to either a clean cotton swab or a swab containing FHVS. We found that, compared to animals exposed to a clean cotton swab, both prepubertal and adult males exposed to FHVS have a greater amount of Fos-immunoreactivity within several brain nuclei comprising the neural circuit mediating male reproductive behavior. Furthermore, this Fos response was equivalent in the two age groups. These results suggest that the inability of the prepubertal male hamster to perform the full repertoire of male reproductive behaviors is not due to a lack of a neuronal activation in response to the pheromonal cues present in FHVS. PMID- 9735227 TI - Expression of testosterone conditioned place preference is blocked by peripheral or intra-accumbens injection of alpha-flupenthixol. AB - Previous evidence indicates that peripheral and intranucleus accumbens injections of testosterone have rewarding effects in male rats as measured in a conditioned place preference (CPP) paradigm. The present study investigated the neurochemical bases of the rewarding properties of testosterone by examining the effect of peripheral and intranucleus accumbens injection of the dopamine receptor antagonist alpha-flupenthixol on expression of testosterone-induced CPP. On alternating days, adult male Long-Evans rats received peripheral injections of testosterone in a water-soluble hydroxypropyl-beta-cyclodextrin (HBC) inclusion complex (0.8 mg/kg) or saline-HBC immediately prior to being confined for 30 min to one of two compartments of a place preference apparatus. All rats received 8 days of pairings (four hormone pairings, four saline pairings). On day 9 the rats were given a 20-min test session during which they had access to all compartments of the apparatus. No hormone was injected prior to the test session; however, rats received a peripheral (20 min prior; 0.2, 0.3 mg/kg) or intra-accumbens (2 min prior, 5.0 micrograms) injection of alpha-flupenthixol or saline. On the test day, rats receiving saline injections spent significantly more time in the compartment previously paired with injections of testosterone than in the compartment previously paired with vehicle injections. In contrast, rats receiving peripheral or intra-accumbens alpha-flupenthixol injections did not spend significantly more time in the compartment previously paired with testosterone. The blockade of testosterone CPP was not due to an effect of alpha flupenthixol on motor behavior. The findings provide further evidence of the rewarding affective properties of testosterone and indicate that peripheral administration and intra-accumbens administration of alpha-flupenthixol block expression of testosterone CPP. The rewarding affective properties of testosterone are mediated, at least in part, via an interaction with the mesolimbic dopamine system. PMID- 9735229 TI - Cholecystokinin induces Fos expression in the brain of the Japanese quail. AB - Systemic administration of cholecystokinin (CCK) inhibits feeding in birds. However, the signaling pathway through which CCK induces this effect is unknown, and its role as a natural satiety signal is controversial. To address these issues, we used immunocytochemistry for the immediate-early gene protein Fos to localize sites of neuronal activation in the brain of Japanese quail (Coturnix japonica) after CCK treatment. Food intake was inhibited in a dose-dependent manner following intraperitonal (i.p.) injection of CCK, with an effective dose range of 1-50 micrograms/kg. To test the hypothesis that CCK induces a distinct pattern of Fos-like immunoreactivity (FLI) in the brain, we compared FLI in birds given CCK (20 micrograms/kg, i.p.) with that in birds given a nonspecific chemical inhibitor of feeding, lithium chloride (LiCl, 40 mg/kg, i.p.), at doses that reduce feeding to a similar level (30% of saline controls). FLI-positive cell nuclei were counted in 14 brain regions after administration of CCK, LiCl, or saline. CCK uniquely induced FLI in the paraventricular, infundibular, periventricular hypothalamic, and medial mamillary nuclei of the hypothalamus. However, CCK and LiCl both induced a comparable pattern of FLI in the hindbrain, with strong staining in the nucleus of the solitary tract and dorsal motor nucleus of the vagus. These findings demonstrate the ability of CCK to activate the central nervous system in birds and suggest that the peptide exerts specific actions in the hypothalamus. However, the possibility that the FLI observed may have arisen through nonspecific effects of CCK on gastrointestinal physiology cannot be discounted. PMID- 9735230 TI - Territorial aggression and dawn song are modulated by septal vasotocin and vasoactive intestinal polypeptide in male field sparrows (Spizella pusilla). AB - Previous experiments demonstrate that lesions of the septum produce opposite effects on intraspecific male aggression in the territorial field sparrow (Spizella pusilla) and the colonial zebra finch (Taeniopygia guttata; facilitate vs inhibit, respectively) and intraseptal infusions of arginine vasotocin (AVT) and vasoactive intestinal polypeptide (VIP) modulate aggression in the male zebra finch (facilitate and inhibit, respectively). The present experiments were conducted to test the hypotheses that (1) septal AVT and VIP modulate both overt territorial aggression and the production of territorial song during the dawn chorus in male field sparrows and (2) these neuropeptides will exert effects opposite of those observed in the zebra finch, consistent with the prediction that social organization is associated with septal neuropeptide function. Wild caught male field sparrows were fitted with chronic guide cannulae directed at the septum and were tested in outdoor aviaries placed in their natural habitat. Intrusion tests (introduction of a stimulus male) and dawn song observations were conducted following infusion of AVT, VIP, or saline control. Consistent with predictions, infusion of AVT significantly inhibited chases and significantly increased chase latency. No significant effects of VIP on chasing or chase latency were observed, although most subjects were more aggressive following infusion of VIP. Both AVT and VIP produced significant, selective effects on the complex (agonistic) song type (facilitation and inhibition, respectively) and produced no effect on the simple (multipurpose) song type. Thus, song and overt aggression appear be modulated independently by septal neuropeptides, and septal AVT and VIP function may differ between species which differ in the expression of territorial or colonial social organizations. PMID- 9735232 TI - In vivo analysis of microvascular injury after myocardial cryothermia. AB - We studied microvascular injury after myocardial cryothermia in rats using intravital fluorescence microscopic techniques. Cryolesions were induced to the right ventricle by freezing with -160 degrees C (probe diameter: 5 mm) for a total of 5 min. Fluorescence microscopy was performed at 15, 30, 60, 90, and 120 min as well as at 3 and 7 days after cryothermia. Analysis of the epicardial microvasculature 15 min after cryothermia revealed an area of 24.6 +/- 3.8 mm2 of nonperfused tissue, which was reduced to 5.3 +/- 1.5 mm2 (P < 0.05) after the initial 2-h observation period. Vital microscopic images of reperfused tissue characteristically demonstrated extravasation of the macromolecular fluorescent tracer FITC-dextran (21.7 +/- 3.4 mm2), suggesting substantial loss of endothelial integrity. In vivo propidium iodide staining confirmed membrane damage of microvascular endothelial cells. Three days after cryoinjury the area of nonperfused tissue was reduced further to 1.1 +/- 0.4 mm2 in the center of the lesion, while the area of perfused tissue with disruption of endothelial integrity was found significantly increased to 47.4 +/- 5.9 mm2 (P < 0.05) toward the periphery. Analysis at 7 days revealed endothelial repair at the periphery of the cryolesion, but now a central necrotic area was found demarcated (nonperfused), presenting with a size (26.0 +/- 3.5 mm2) similar to that shown during the very early (15 min) reperfusion period. Our study demonstrates recovery of microvascular perfusion during the first hours and days after myocardial cryothermia. This is, however, associated with endothelial injury, i.e., damage of plasma membrane and loss of barrier function. Infarction with capillary perfusion failure is evident at 7 days with a size which strikingly corresponds to the sizeof nonperfused tissue observed immediately after cryointervention. PMID- 9735233 TI - Basic fibroblast growth factor stimulates angiogenesis in the hindlimb of hyperglycemic rats. AB - Angiogenic growth factors including basic fibroblast growth factor (bFGF) have therapeutic value for chronic ischemia in nondiabetic animals. However, angiogenic therapy for chronic ischemia in a background of diabetes remains unexplored. In the present study, we evaluated the effects of exogenous bFGF on angiogenesis in streptozotocin-induced diabetic rats with ischemic and nonischemic limbs. We produced ischemia of the left lower limb by excising the superficial femoral artery. At 2 weeks, the rats received an intramuscular injection of vehicle (group A), 0.3 microg bFGF/day (group B), or 1 microg bFGF/day (group C), daily for 2 weeks. At 4 weeks, we assessed limb angiogenesis by skeletal muscle capillary density (cap/mm2) and capillary per muscle fiber ratio (cap/F) counts. Group C had significantly higher mean levels compared to group A for calf capillary density (P < 0.0024) and capillary per muscle fiber ratio in both thigh (P < 0.0015) and calf (P < 0.0001). There was a trend toward increased mean capillary per muscle fiber ratio with increasing dose. This trend was significant in the calf (P < 0.0015) and just missed statistical significance in the thigh. There was a similar trend in calf capillary density. We conclude that exogenous bFGF enhances angiogenesis and, possibly, collateral circulation in ischemic limbs of diabetic rats. PMID- 9735234 TI - In vitro influences between pancreatic adenocarcinoma cells and pancreatic islets. AB - BACKGROUND: Interactions have been found between exocrine pancreatic adenocarcinoma and islets of Langerhans. Growth of pancreatic adenocarcinoma cells can be regulated by islet hormones such as insulin and somatostatin. Conversely, dysfunction of endocrine pancreas frequently accompanies the exocrine malignancy. The mechanisms underlying these interactions have not been defined. MATERIALS AND METHODS: Human pancreatic adenocarcinoma cells (HPAF cells) were cocultured with isolated rat pancreatic islets in two-compartment wells. HPAF cells and islets cultured in separate wells served as controls. In separate experiments, HPAF cells were incubated with two concentrations of exogenous insulin, including one reflecting the levels of insulin secretion seen in the coculture experiments. RESULTS: Proliferation of HPAF cells was increased by about 50% following a 2- or 5-day incubation with pancreatic islets (P < 0.05). Coculture of HPAF cells and pancreatic islets was associated with a greater reduction in glucose concentrations (P < 0. 01) and an increase in lactate accumulation (P < 0.05) in the culture media. Insulin concentrations in the media were significantly decreased during the first 2-3 days of the coculture incubation (P < 0.05). In contrast, insulin secretion from control islets was not significantly decreased until the fifth day of the experiment. The growth of HPAF cells was stimulated by both concentrations of exogenous insulin (P < 0.05). The insulin-stimulated HPAF cells also showed an enhanced glucose consumption and lactate production (P < 0.05). CONCLUSIONS: Pancreatic islets regulate both growth and glucose metabolism of adjacent exocrine cancer cells. beta-cell derived insulin may be one of the factors inducing these effects. Insulin release from islet beta-cells is compromised in the presence of exocrine cancer cells. PMID- 9735235 TI - Apoptosis in esophageal cancer following induction chemoradiotherapy. AB - BACKGROUND: The poor survival of patients with esophageal cancer following esophagectomy has led to intense investigation into combined modality therapy. Based on results from clinical trials examining chemoradiotherapy alone without surgery, resection has come under increased scrutiny and its necessity as a component of a multimodal approach has been questioned. In this study, we examined whether residual tumor cells in esophagectomy specimens following induction chemoradiotherapy are viable and, therefore, provide putative evidence for the appropriateness of esophagectomy. MATERIALS AND METHODS: Between August 1991 and January 1995, 46 patients were entered into an induction chemoradiotherapy trial consisting of 5-fluorouracil, cisplatin, alpha interferon, and concurrent external beam radiotherapy followed by esophagectomy. Response was determined histologically and apoptosis assessed with a terminal deoxytransferase assay system. p53 status was determined by immunohistochemistry and mutational analysis. RESULTS: Thirty-eight patients underwent esophagectomy, 33 of whom had either a complete (n = 10) or partial (n = 23) response. None of the 28 patients with residual tumor in the resected specimen had 100% apoptotic cells and the vast majority of specimens had less than a 10% apoptotic rate. The percentage of apoptotic cells did correlate with tumor differentiation but not with histologic type nor presence of p53 mutations. CONCLUSIONS: These data suggest that resection following upfront chemoradiotherapy is a necessary component of a multimodality approach to esophageal cancer and will ultimately provide superior local-regional control to a nonsurgical approach. PMID- 9735236 TI - Patellar tendon and infrapatellar fat pad healing after harvest of an ACL graft. AB - Clinical studies have documented proliferation of the host patellar tendon and fibrosis extending into adjacent tissues after reconstruction of the injured anterior cruciate ligament (ACL) using the central one-third of the patellar tendon (PT) as the graft. Such generalized arthrofibrosis has been implicated in knee locking and as possible source of anterior knee pain. However, it is not clinically feasible to measure changes in tendon morphology and mechanical properties and degeneration of peripheral tissues over time following graft harvest. In a rabbit experimental model proliferative changes in the tendon and the infrapatellar fat pad have been documented following harvest of a central third tendon graft without ACL reconstruction. Studies in larger animals have shown significant reductions in the strength and stiffness of the healing patellar tendon, but without assessment of the peripheral tissue response. In the current study an ACL reconstruction was performed in a goat model using an autogenous patellar tendon graft. Extensive tendon and fat pad proliferation were observed along with significant reductions in the biomechanical properties of the host tendon. Significant fat pad fibrosis was documented using biochemical methods. The current data confirm that harvest of an autogenous PT graft for reconstruction of the ACL results in significant changes in the PT and adjacent tissues. These data may help explain some of the clinical complications documented in the reconstructed joint. PMID- 9735237 TI - Immunohistochemical analysis of clinically transplanted muscles. AB - BACKGROUND: Although a number of studies have examined the morphology and function of experimentally transplanted muscles, immunohistochemical evaluation of clinically transplanted muscles has not been reported. The purpose of this study was to examine clinically transplanted muscles at long periods after transplantation with biochemical markers specific for satellite cell activation and muscle regeneration. MATERIALS AND METHODS: Nine biopsies of muscles transplanted to the paralyzed face were examined. In five cases, the gracilis muscles were transplanted about 1 year after cross face nerve grafting. The other four cases underwent one-stage latissimus dorsi (LD) muscle transplantation. Twelve to 162 months after transplantation, muscle biopsies were harvested in nine cases. In eight cases, secondary corrections of facial expression including debulking of the grafted muscle were required, while another muscle was transplanted in one case because of the failed first operation. As control, six specimens of normal LDs were examined as well. Monoclonal antibodies were employed to visualize myosin heavy chain (MHC) isoforms (slow, fast, and embryonic) and MyoD protein. RESULTS: Although one specimen exhibited only small, atrophic fibers indicating failed reinnervation, the remaining eight specimens showed regularly distributed fibers and type grouping indicating successful reinnervation. There was no statistically significant difference in fiber area and lesser diameter between normal LDs and transplanted LDs. However, even in these successfully reinnervated muscles, intermediate and small fibers expressing embryonic MHC and small cells expressing MyoD were observed, suggesting that satellite cells were activated for repair of the adjacent fibers. CONCLUSIONS: Muscle adaptation (presumably to denervation), which is a regenerative change accompanied by activation of satellite cells, was still seen even long periods after transplantation. It is concluded that, in microneurovascular human skeletal muscle transfers, there is a wide variation in the time required for reinnervation of individual muscle fibers, and it may be that human muscle fibers cannot be properly reinnervated after denervation has continued for a certain period such as 12 months. PMID- 9735238 TI - Increased gut permeability after hemorrhage is associated with upregulation of local and systemic IL-6. AB - Although intestinal barrier failure after hemorrhage is a well-documented event, the underlying mechanism is poorly understood. The aim of this study, therefore, was to determine whether altered intestinal permeability after hemorrhage is associated with upregulation of local and systemic interleukin-6 (IL-6). To study this, rats underwent laparotomy (i.e., trauma induced) and were bled to and maintained at a mean arterial pressure of 40 mm Hg until 40% of the shed blood volume was returned in the form of Ringer's lactate. The animals were then resuscitated with four times the volume of shed blood with Ringer's lactate over 60 min. At 1.5 h postresuscitation, an in vivo ligated loop of a distal small intestine was formed and the passage of 4-kDa fluorescein isothiocyanate conjugated dextran (FD4) from the intestinal lumen into the portal vein and carotid artery blood was analyzed by fluorescence spectrometry. Samples from the portal vein and a carotid artery were collected and plasma IL-6 was assayed. Intraepithelial lymphocytes from a distal small intestine were isolated and cultured in vitro for 24 h with or without anti-rat CD3 monoclonal antibody stimulation. IL-6 activity in freshly isolated cells and its release by cultured lymphocytes were determined. Intestinal perfusion and portal blood flow were determined by radioactive microspheres in another set of parallel experiments. The results indicate that lumen-to-blood passage of FD4 through the wall of the small intestine increased significantly at 1.5 h after hemorrhage and resuscitation and was associated with decreased intestinal perfusion and portal blood flow. Plasma IL-6 levels in the portal vein and carotid artery markedly increased at 1.5 h after hemorrhage and resuscitation. In addition, a significant correlation was observed between plasma IL-6 and FD4 concentrations. Higher IL-6 activity in freshly isolated cells was found in hemorrhaged rats. Increased IL-6 release by cultured lymphocytes was also observed either with or without anti-rat CD3 monoclonal antibody stimulation. Thus, the increased intestinal permeability following trauma-hemorrhage and resuscitation appears to be associated with systemic and intestinal IL-6 upregulation. PMID- 9735239 TI - Heat shock preconditioning ameliorates liver injury following normothermic ischemia-reperfusion in steatotic rat livers. AB - The decreased tolerance of steatotic livers to warm ischemia complicates liver surgery. The efficacy of heat shock preconditioning in steatotic livers to lessen ischemia-reperfusion injury was studied in rats. Steatotic liver was produced in Lewis rats with a choline-deficient diet. Rats with steatotic livers were divided into a heat shock preconditioned group (group HS) and a control group (group C). All rats received 45 min of hepatic warm ischemia. Survival rates and changes in biochemical and histological parameters were compared in both groups. Heat shock protein 72 (HSP72) was produced only in group HS. The 7-day survival of the rats after warm ischemic intervention was significantly better in group HS (13/15) than in group C (5/15) (P < 0.01). The concentration of ATP in liver tissue (n = 10, P < 0.01) and serum levels of aspartate aminotransferase (n = 10, P < 0.05), alanine aminotransferase (n = 10, P < 0.01), and lactic dehydrogenase (n = 10, P < 0.01) at 40 min reperfusion were also significantly better in group HS than in group C. Histological examination at 40 min reperfusion showed severe sinusoidal congestion, hepatocyte necrosis, and increased positivity to 4-hydroxy-2-nonenal modified proteins in group C livers; these signs were markedly suppressed in group HS livers. The data indicate that heat shock preconditioning provides the steatotic rat liver with significant tolerance to warm ischemia-reperfusion injury. PMID- 9735240 TI - In vitro morphological and functional characterization of isolated porcine hepatocytes for extracorporeal liver support: bile acid uptake and conjugation. AB - Recently, researchers have focused on the use of bioartificial liver (BAL) to support patients with fulminant hepatic failure (FHF). We have developed a cell based BAL, consisting of porcine hepatocytes in a hollow-fiber bioreactor. To better characterize BAL metabolic functions in vitro, bioreactors were inoculated with 48-h-cultured, microcarrier-attached hepatocytes and perifused with recirculating human plasma that contained either 1 microCi of [24-14C] plasma enriched cholate or 1 microCi of [24-14C] plasma-enriched taurocholate. Bile acids were sampled hourly and separated into four fractions (unconjugated, glycoconjugated, tauroconjugated, and sulfated) for radioactivity determination. Following 3 h perifusion, the glycoconjugated and sulfated bile acid fractions in the bioreactor extrafiber space were significantly elevated when compared to the recirculating plasma. During perifusion with taurocholate-enriched plasma, a relative decrease in the tauroconjugated fraction and an increase in the glycoconjugated fraction were observed. Cholate was accumulated by hepatocytes to a level threefold lower than taurocholate; however, a significant proportion of radioactivity (<25%) was detected in the glycoconjugated fraction. Ultrastructural examination of microcarrier-attached hepatocytes illustrated that the features typical of metabolically active liver cells were maintained. Our data demonstrate the ability of BAL to clear bile acids from the circulation, to accumulate cholate and taurocholate, and to conjugate a substantial amount of cholic acid. PMID- 9735241 TI - Significance of serum delta-bilirubin in patients with obstructive jaundice. AB - BACKGROUND: Delta-bilirubin is a bilirubin covalently bound with albumin, which is nontoxic and excreted neither in urine nor in bile. We previously reported that the percentage of delta-bilirubin increased after biliary drainage and that the rapidly excretable bilirubin fraction (total minus delta-bilirubin) was a better parameter to predict the effectiveness of biliary decompression in the dog model. The aim of the present study was to elucidate whether it is applicable to humans. MATERIALS AND METHODS: The serum bilirubin concentration was measured and its fractions were analyzed by high-performance liquid chromatography in 22 patients with obstructive jaundice before and after biliary drainage. In addition, the patients were subgrouped into good and poor drainage groups according to the decline index of serum bilirubin to examine the significance of delta-bilirubin. RESULTS: The concentration of total bilirubin decreased from 14.1 mg/dl before biliary drainage to 5.4 mg/dl 28 days after drainage. During this period, the percentage of conjugated bilirubin steeply declined from 47.1 to 8.8% and that of excretable bilirubin from 63.4 to 28.6%. In contrast, the proportion of serum delta-bilirubin increased from 36.6 to 71.4%. There was an inverse correlation between percentage of delta-bilirubin and total bilirubin concentration (r = -0.69, P < 0.01). In the good drainage group, the percentage of delta-bilirubin increased above 60% within 7 days after biliary drainage, but it did not reach 60% by 28 days in the poor drainage group. A decreasing rate of total bilirubin minus delta-bilirubin, the excretable bilirubin fraction, was a better index than that of total bilirubin to assess the efficacy of biliary drainage (P< 0.01). CONCLUSIONS: The increase in the percentage of serum delta bilirubin indicates an effectiveness of biliary drainage in man. An analysis of serum delta-bilirubin for 7 days can distinguish the good drainage patients from the poor drainage patients. PMID- 9735242 TI - Sustained nitric oxide exposure decreases soluble guanylate cyclase mRNA and enzyme activity in pulmonary artery smooth muscle. AB - BACKGROUND: The soluble isoform of guanylate cyclase (sGC) is activated by nitric oxide (NO) to form guanoside 3':5'-cyclic monophosphate (cGMP). Cyclic GMP levels cause smooth muscle relaxation and regulate vascular tone to various vascular beds, including the lung. Under conditions of cytokine excess the inducible synthesis of NO may result in cGMP overproduction, generalized vasodilatation, and septic shock. In the pulmonary bed the opposite response, pulmonary hypertension, may occur. We hypothesized that sGC activity decreases in the face of sustained levels of NO. MATERIALS AND METHODS: We used the NO-donor S-nitroso acetyl-D-L-penicillamine to study the effects of NO on sGC mRNA abundance and enzyme activity in cultured rat pulmonary artery smooth muscle cells. RESULTS: NO caused a prompt rise in extracellular cGMP production. Pretreating cells with NO for >/=45 min inhibited subsequent cGMP synthesis. NO-pretreated cells recovered the capacity for cGMP synthesis after removal of NO for 120 min. When actinomycin or cycloheximide was added to NO pretreatment, cells retained cGMP synthetic capacity. NO pretreatment decreased sGC mRNA abundance, but did not totally eliminate it. CONCLUSION: NO has important regulatory effects on cGMP synthesis at the level of enzyme activity and mRNA abundance. NO causes an immediate synthesis of large amounts of cGMP. With prolongation of exposure (>/=60 min) sGC enzyme activity decreases and cGMP production drops significantly. Soluble GC mRNA abundance also decreases and may result in decreased responsiveness of cells to NO with regard to cGMP production. PMID- 9735243 TI - Multiporous cellulose microcarrier for the development of a hybrid artificial liver using isolated hepatocytes. AB - BACKGROUND: This study was aimed at developing an optimal method for immobilizing isolated hepatocytes in cellulose multiporous microcarriers (MCs) and evaluating the metabolic activity of MC-immobilized hepatocytes. MATERIALS AND METHODS: Hepatocytes isolated from the livers of male Wistar rats were immobilized in collagen-coated MCs by intermittent stirring (30 rpm for 2 min per 15 min) for 180 min or accumulation methods. The accumulation method was performed by pouring aliquots of hepatocyte suspension (8 x 10(5)) and MC suspension (1 mg) in turn onto a nylon mesh (pore size: 100 micron). The metabolic activity of MC immobilized hepatocytes in floating culture and in a newly developed bioreactor was evaluated. The metabolic activity of MC-immobilized hepatocytes in the bioreactor was also evaluated in in vitro perfusion of a hollow-fiber-based hybrid artificial liver support system. RESULTS: The accumulation method immobilized 20 times more hepatocytes in collagen-coated MCs than the intermittent stirring method (P < 0.01). Morphological observation of hepatocyte immobilized MCs revealed that many hepatocytes were immobilized deep within the MCs maintaining a spherical shape and normal microvilli on their surface. MC immobilized hepatocytes in floating culture revealed similar NH3 metabolism and glucose synthesis to monolayer-cultured hepatocytes, and this metabolic activity was maintained during 9h of floating culture. MC-immobilized hepatocytes in a bioreactor also showed similar NH3 metabolism to monolayer-cultured hepatocytes. The NH3 metabolism of MC-immobilized hepatocytes in in vitro perfusion of a hybrid artificial liver support system was 241.5 microg/h/mg protein/m2 membrane surface. CONCLUSIONS: The results of this study indicate that the accumulation method was optimal for immobilizing isolated rat hepatocytes in MCs and that MC immobilized hepatocytes maintained their metabolic activity for a long period. PMID- 9735244 TI - Release of endothelin-1 in strangulation obstruction of the small bowel in pigs. AB - Evidence has been provided that increased portal vein pressure results in increased release of endothelin-1 (ET-1). Strangulation obstruction is associated with increased venous pressure, and we wanted to determine if it is associated with increased local release of ET-1 and elevated concentration of ET-1 in systemic blood. Strangulation obstruction was induced by elevating pressure in a gasket placed around a loop of ileum until venous pressure reached 50 mm Hg. Ischemia in a bowel loop was induced by arterial clamping, reducing blood flow by 70%. Blood samples were collected before and after 30, 90, and 180 min of strangulation or ischemia. ET-1 was determined by radioimmunoassay following acidification and extraction on C18 columns. In strangulated loop the blood flow decreased by 70%. ET-1 concentration remained around 5 pg/ml in arterial blood, increased fourfold in strangulated venous blood, and remained unchanged in venous blood from control bowel. The release of ET-1 from the strangulated loop to blood increased twofold. Ischemia resulted in reduced release of ET-1. It is concluded that strangulation obstruction causes increased release of ET-1 to venous blood in the strangulated loop, but not increased ET-1 concentration in systemic blood. The increased ET-1 release was probably due to increased venous pressure, not to low blood flow. PMID- 9735245 TI - Reperfusion induces sublethal endothelial injury. AB - BACKGROUND: Endothelial cells are pivotal in regulating thrombosis and hemostasis. In this study, we sought to characterize endothelial dysfunction and endothelial cell injury in vitro after hypoxia/reoxygenation. MATERIALS AND METHODS: Cultured human umbilical vein endothelial cells (ECs) were exposed to 120 min of hypoxia followed by reoxygenation. The release of thrombomodulin (TM) and the production of prostaglandin I2 (PGI2) were measured. Endothelial cell injury in hypoxia/reoxygenation was measured by two assays, the Fura-2 release assay and the 51chromium (51Cr) release assay. RESULTS: TM release from ECs during normoxic incubation was undetectable, while it was slightly increased during hypoxic incubation. After reoxygenation, the release of TM increased, and it became significantly higher at 120 min after reoxygenation compared with hypoxic incubation. The production of PGI2 significantly decreased during hypoxic incubation and further decreased within 30 min after reoxygenation, but returned to normoxic levels at 120 min after reoxygenation. In the Fura-2 release assay, a rapid and significantly greater release of Fura-2 was observed in hypoxia/reoxygenation compared with hypoxic incubation. In the 51Cr release assay which demonstrates cell death, 51Cr release did not increase in hypoxia/reoxygenation. CONCLUSIONS: The present study suggests that 120 min of hypoxia/reoxygenation induces endothelial dysfunction of ECs but does not cause cell death. PMID- 9735246 TI - Evolutionarily Stable Reproductive Strategies in Sexual Organisms: IV. Parent Offspring Conflict and Selection of Seed Size in Perennial Plants. AB - The provisioning of offspring in sexually reproducing organisms provides an arena in which genetic conflict of interests between parents and their offspring may be expressed. While most existing models of parent-offspring-conflict consider the case of a parent that rears one offspring a year, this paper is concerned with perennial plants that produce many seeds at one time. Parent-offspring conflict is examined in the context of an integrated analysis of reproductive allocation, sex allocation, and the amount of resources invested in each offspring. I derive the evolutionarily stable strategy (ESS) results for the allocation of resources when the mother plant is in control as well as when the offspring are in control taking into account both density-independent and density-dependent population growth. To the extent that the relationships between gamete output and resource investment are linear for both sex functions, the separate treatment of reproductive effort, sex allocation, and offspring size-number compromise in modern life-history theories is justified, regardless of which side, parents or offspring, wins the conflict. In such cases, the ESS sex allocation is exactly what is found in traditional sex allocation theory, and the ESS reproductive effort maximizes the rate of population growth in density-independent populations, or the amount of resources allocated to reproduction during an average lifespan in density-dependent populations. In contrast to the previous theoretical conclusions based on the analyses of single-offspring cases, the ESS reproductive effort under the offspring's control of allocation to individual offspring is found to be lower than that when mothers are in control. This paradoxical result occurs because a mother producing fewer ovules fares better if she knows that each of her seed offspring will get more resources than the amount she is selected to give. The evolutionarily stable offspring size in both density independent and density-dependent populations does not depend on mother's reproductive effort and investment-independent mortality, just as traditional models of offspring size-number trade-offs would predict.Copyright 1998 Academic Press Limited PMID- 9735247 TI - Extending the Iterated Prisoner's Dilemma without Synchrony. AB - In biology altruistic behaviour of selfish individuals often can be modelled by the iterated Prisoner's Dilemma game (PD). If the opponents are caught repeatedly in this dilemma reciprocity may lead to cooperative strategies adopted by the individuals. In this article we present results from numerical simulations of the infinitely iterated stochastic alternating PD. First we investigate influences of the memory size on strategies in the alternating two player PD. We show that Firm but Fair is a strong strategy hardly affected by memory size and different values of the temptation to defect. Second we discuss successful strategies in the alternating N player N step memory PD. In this situation we focus on the stability of cooperative strategies and compare the results to experiments of predator inspection by sticklebacks carried out by Milinski and co workers.Copyright 1998 Academic Press Limited PMID- 9735248 TI - Reconstructing the Evolution of Viviparity and Placentation. AB - A recent paper in J. theor. Biol. has challenged the proposed application of punctuated equilibrium models to the evolution of reptilian viviparity and placentation. While clarifying some aspects of the models, the paper's criticisms reflect misinterpretations of the literature and an unnecessary reluctance to apply punctuationist concepts to extant taxa. The punctuated equilibrium model retains potential for clarification of the patterns of stasis and episodic change that may well have characterized squamate reproductive history.Copyright 1998 Academic Press Limited PMID- 9735249 TI - Dynamics of Cytoplasmic Incompatability with Multiple Wolbachia Infections. AB - Wolbachia infections occur in many arthropods. These matrilineally inherited bacteria cause cytoplasmic incompatibility, in which a cross produces no offspring when between an infected male and an uninfected female. Some populations harbour multiple Wolbachia strains. Females fail to produce offspring when crossed to a male with a strain that the female lacks. Prior theoretical work showed that a panmictic population cannot maintain polymorphism for different strains when each female carries only a single strain. A few authors suggested that doubly infected females can stabilize multistrain polymorphism, but conditions for invasion and location of stable equilibrium were not analysed in detail. For two strains, I describe the conditions under which a multiply infected class can spread. Spread of the doubly infected type stabilizes polymorphism of the singly infected classes. This analysis also suggests an interesting extension to higher multiplicity of infection. For an arbitrary number of strains, N, a panmictic population cannot maintain different classes with N-1 infections unless the class with N infections is also present. This pyramid of polymorphism may explain the puzzling diversity of incompatibility types observed in some Culex mosquitos. Multiple infection also has interesting consequences for the dynamics of spatial variation and reproductive isolation.Copyright 1998 Academic Press Limited PMID- 9735250 TI - A New Model for the Molecular Mechanisms of Photosynthetic Water Oxidation and Photophosphorylation. AB - It is postulated that trans-3-phosphatidyl glycerol, tightly bound to the inner side of the thylakoid membrane, catalyses-after its oxidation to the oxygen radical by P680 in combination with the tyrosine radical YZ. and after release of one proton-in cooperation with the Mn enzyme the first reaction in water splitting. In this way, four molecules of water would be oxidized by four light flashes. In the last phase the Mn enzyme would act as a "catalase" transforming four "complexed OH species" (2H2O2) to 2H2O+O2. The photophosphorylation is formulated analogously to the mitochondrial process, because the structure of the ATP synthase in chloroplasts on principle agrees with the mitochondrial enzyme complex. Cardiolipin or cardiolipin ketone, respectively, may be exchanged by tightly-bound phosphatidyl glycerol or glycerone, respectively. Accordingly, to enable ATP synthesis in purified in vitro systems (MF0F1 or CF0F1 ATP synthase), a redox reaction or light energy for formation of the ketyl radical and an H+/Na+ gradient are necessary. SH compounds, valinomycin-K+, carbonyl cyanide m chlorophenylhydrazone (CCCP), organic acids, or cholesterol are suitable as electron donors. Moreover, it is postulated that MgATP, synthesized by the catalytic centres of the F1 part, is shifted to the allosteric nucleotide-binding sites to elevate-before its release-the MgADP affinity of the respective following catalytic centre. In this way, the synthesis product MgATP is additionally used as an allosteric effector, before it is released for energy yielding reactions. So the ATP synthesis can proceed in an optimal rhythm.Copyright 1998 Academic Press Limited PMID- 9735251 TI - A Life-history Invariant for Migration. AB - Copyright 1998 Academic Press Limited PMID- 9735252 TI - Evolutionarily Stable Reproductive Strategies in Sexual Organisms. Part V-Joint Effects of Parent-offspring Conflict and Sibling Conflict in Perennial Plants. AB - We develop a general treatment of the joint effects of parent-offspring conflict (conflict between broods) and sibling conflict (conflict within broods) in perennial plants. Parent-offspring conflict as well as sibling conflict are examined in the context of an integrated analysis of reproductive allocation, sex allocation, and the amount of resources invested in each offspring. We find that under a wide range of conditions the selected seed size is independent of reproductive effort or sex allocation. To the extent that the relationships between gamete output and resource investment are linear for both sex functions, the separate treatment of reproductive effort, sex allocation, and offspring size number compromise in modern life-history theories is well justified, regardless of which side, parents or offspring, controls the allocation to individual seed offspring. We show that incorporation of sibling conflict results in even larger seed size, which in turn leads to even lower reproductive effort, than under pure parent-offspring conflict. If the costs of increased provisioning of selfish offspring are entirely borne by their brood mates, then offspring are selected to solicit parental care as long as they continue to benefit individually from more resources.Copyright 1998 Academic Press Limited PMID- 9735253 TI - On a Wing and a Vector: a Model for Magnetic Navigation by Homing Pigeons. AB - The ability of pigeons to home directly from distant, unfamiliar release sites has defied explanation because it has been impossible to identify the mechanism by which the birds determine their current position relative to their home loft. A variety of magnetic effects on homing orientation have implicated magnetic total intensity in position determination but no testable models for magnetic navigation by homing pigeons have resulted. Here, a vector summation model is proposed which identifies a novel coordinate that pigeons could use with magnetic total intensity to determine position. The model makes predictions about the accuracy of homing and patterns of homing orientation over local and regional scales. The model requires no unusual computational or cognitive abilities. It is, however, consistent with a significant volume of pigeon homing data and can be tested in a variety of ways.Copyright 1998 Academic Press Limited PMID- 9735254 TI - System-dependent Selection, Ecological Feedback and the Emergence of Functional Structure in Ecosystems. AB - Most models of natural selection assume either that the material environment remains constant or that it fluctuates in ways unrelated to changes in gene frequencies (and therefore changes in the distribution of phenotypes) of the organism undergoing selection. In this paper, we consider what happens when this assumption does not hold, that is, when ecological feedback between organism and environment is included in the evolutionary process. Specifically, we examine the unusual evolutionary dynamics that occur when changes in the distribution of phenotypes (resulting from selection) alter an environmental parameter in ways that, in turn, modify selection pressures. This process, which we term "system dependent selection", can produce stable phenotypic diversity which functions to regulate the relevant environmental parameter within a much narrower range would occur in the absence of ecological feedback. This environmental regulation raises the mean fitness of the population and reduces variance in fitness among different phenotypes. Thus, system-dependent selection produces functional organization at the level of the system, as a whole, rather than at the level of the individual organism. We use James Lovelock's model of the imaginary planet Daisyworld to describe the unusual dynamics of this selective process and then use a similar model to examine the structure of an ancient system of wet-rice farming on the Indonesian island of Bali. This model accurately predicts the actual structure of functional organization along two Balinese rivers. We investigate the stability of such systems by exploring the conditions under which mutant phenotypes can invade Daisyworld. The results suggest that the phenotypic diversity and functional organization produced by system-dependent selection may be maintained when there exists variation, over evolutionary time, in the environmental parameters underlying system-dependent dynamics.Copyright 1998 Academic Press Limited PMID- 9735255 TI - A Theoretical Analysis of Sea-anchor Soaring. AB - This study presents a concise mathematical model that accounts for sea-anchor soaring, a special flight technique used by sea birds, in particular storm petrels. Conventional wing theory is used to reveal the mechanics of sea-anchor soaring. The feasibility and existence of an equilibrium are summarized by formulae giving the wind velocity criteria. The stability of the equilibrium is also revealed: among two possible equilibria, the equilibrium at the very low velocity to water is shown to be stable. Numerical examples show the following: sea-anchor soaring is almost always stable; Wilson's storm petrel can soar at 0.256 ms-1 in wind with 4.79 ms-1 under the best conditions. Larger sea-birds like albatrosses need gusty winds to soar, but such strong winds make the sea choppy and hence prevent soaring. Foot-web size regulates the size of a bird utilizing sea-anchor soaring at low velocities to water.Copyright 1998 Academic Press Limited PMID- 9735256 TI - Comparison of Stochastic and Deterministic Concepts of Bacterial Lag. AB - The shortcomings of the traditional deterministic approach to bacterial lag are exposed in this paper. A more precise, stochastic, formulation is put forward which takes account of the variation of the individual cell's lag time. Formulae are given to describe how the lag-distribution of the cells relates to the deterministic population lag commonly used as a practical measure of bacterial lag time.Copyright 1998 Academic Press Limited PMID- 9735257 TI - On Units of Selection in Cultural Evolution. AB - Copyright 1998 Academic Press Limited PMID- 9735258 TI - Migration Dynamics of Fish Schools in Heterothermal Environments. AB - A mechanistic theory is presented to illustrate the ecologically scaled behavior of highly organized fish schools, which is principally controlled by thermal structure of the habitat. Starting from the equation governing the schooling dynamics of fish, and incorporating the hypothesized memory mechanism of thermal history into the model of klinokinesis, a mathematical formalism is developed which relates the individual processes of perception of temperature-gradients to the thermoregulatory migration of schools. The migration dynamics of schools in heterothermal environments is formulated as the modified Fick's law of diffusion in the presence of external potential field, which gives the advection-telegraph equation in combination with the continuity equation. A behavioral basis for the environmental potential function is proposed, which has been postulated ad hoc as a driving force of drift velocity in modeling biological aggregation patterns. It is predicted that the distribution of schools self-organizes into a critical state.Copyright 1998 Academic Press PMID- 9735259 TI - Homogeneous and Spatio-temporal Chaos in Biochemical Reactions With Feedback Inhibition. AB - A biochemical oscillator involving feedback inhibition can be extended to generate a chaotic attractor. The simple chaotic attractor may be exploited to create signals of higher dimension by passive forcing of other reactions. Diffusively coupled oscillators with negative feedback exhibit regular and irregular patterns in time and space associated with periodic and hyperchaotic attractors, respectively.Copyright 1998 Academic Press PMID- 9735260 TI - Curvature of Swimming Fish Midlines as an Index of Muscle Strain Suggests Swimming Muscle Produces Net Positive Work. AB - The axial muscle used in steady swimming by fish is geometrically complex and this has required the study of in vivo muscle swimming mechanics to be done with largely inferential techniques. Currently there is some debate concerning the variation in muscle function in different regions of the body, and the importance of negative work production by muscles in posterior locations. We have used video taped kinematics of steady swimming in mackerel and then analysed the lateral flexion of the body that is thought to reflect the cycle of length change in the axial muscles. A comparison of several of the techniques used in the past to estimate muscle length changes are shown not to be equivalent. Specifically, we find that peak values of lateral deflection of the body are not correlated in time or axial position with peaks in curvature of the body midline. This is important because the timing of the muscle strain cycle is often inferred from such information. Having documented this observation in a swimming mackerel, we use analytical geometry to show that this result is a consequence of the curves that describe swimming fish midlines. Since this observation is shown to be the geometric consequence of an amplitude envelope that is not constant, it should therefore apply to other animals that propel themselves with axial undulations of increasing amplitude. Our minor result is that care must be taken when estimating the phase of swimming muscle strain from kinematics. Our major results is that appreciation of the geometric character of the kinematics these fish adopt may resolve much of the current debate concerning the mechanical performance of swimming muscle in fish. Specifically, using our estimate of the phase of muscle shortening and data on EMG timing in the mackerel we conclude that all axial muscles are activated before peak length and in a manner that should produce net positive work within each shortening cycle.Copyright 1998 Academic Press PMID- 9735261 TI - The Role of Weak Interactions in Biological Systems: the Dual Dynamics Model. AB - The dual dynamics model is a random autonomous network of nodes whose dynamical behavior is determined by both strong and weak interactions. The model combines discrete decision-making features reflective of logical operations with arithmetic features that represent graded influences. Dual dynamics abstracts the ubiquitous fact that biological systems at all levels of organization consist of components that respond both to specific (strong) signals and to the cumulative effect of numerous weak interactions. We have thoroughly studied the dynamical characteristics of three-valued dual dynamics networks in the range from four to 14 nodes and have compared these characteristics to those of non-boolean three valued networks (without weak interaction) and to Kauffman boolean networks. Properties studied include: attractor length, number of attractors, basin sizes, orbital stability, and evolutionary transformability in response both to individual and cumulative mutations (where mutations are implemented as random changes in the response of a node to the pattern of strong influences impinging on it). The introduction of weak interactions and their manner of coupling to strong interactions has major altering effects on these properties. With suitable coupling it is possible to significantly enhance equifinality and evolutionary transformability. The model demonstrates that self-organizing dynamics are compatible with evolutionary plasticity when weak interactions are taken into account.Copyright 1998 Academic Press PMID- 9735262 TI - A Morphogenetic Model Accounting for Pollen Aperture Pattern in Flowering Plants. AB - Pollen grains are embeddded in an extremely resistant wall. Apertures are well defined places where the pollen wall is reduced or absent that permit pollen tube germination. Pollen grains are produced by meiosis and aperture number definition appears to be linked with the partition that follows meiosis and leads to the formation of a tetrad of four haploid microspores. In dicotyledonous plants, meiosis is simultaneous which means that cytokinesis occurs once the two nuclear divisions are completed. A syncitium with the four nuclei stemming from meiosis is formed and cytokinesis isolates simulataneously the four products of meiosis. We propose a theoretical morphogenetic model which takes into account part of the features of the ontogeny of the pollen grains. The nuclei are considered as attractors acting upon a morphogenetic substance distributed within the cytoplasm of the dividing cell. This leads to a partition of the volume of the cell in four domains that is similar to the observations of cytokinesis in the studied species. The most widespread pattern of aperture distribution in dicotyledonous plants (three apertures equidistributed on the pollen grain equator) can be explained by bipolar interactions between nuclei stemming from the second meiotic division, and observed variations on these patterns by disturbances of these interactions. In numerous plant species, several pollen grains differing in aperture number are produced by a single individual. The distribution of the different morphs within tetrads indicates that the four daughter cells can have different aperture number. The model provides an explanation for the duplication of one of the apertures of a three-aperture pollen grain leading to a four aperture one and in parallel it gives an explanation for how heterogeneous tetrads can be formed.Copyright 1998 Academic Press PMID- 9735263 TI - War of Attrition with Individual Differences on RHP. AB - The fact that there always exists various kinds of almost continuous mutations for any animal population implies that players in competitions can never be perfectly symmetric in any sense. To develop a model to fit this reality, we consider war of attrition games in which players have continuously different resource holding potential (RHP). The RHP of each opponent is not known in our settings. Pure ESS functions and Nash equilibria are obtained under sufficiently rotational conditions as unique solutions of certain differential equations among the class of Lebesgue measurable functions. They are normal in that a higher RHP induces a longer attrition time, which implies that a player with greater RHP always wins. This model includes as the limit the conclusions of Maynard Smith (1974, J. theor. Biol. 47, 209-221) and Norman et al. (1977, J. theor. Biol. 65, 571-578), which did not consider individual differences in RHP. Our results suggest that, by changing each player's qualitative differences to continuous quantitative differences, some of the mixed ESS solutions previously found in discrete games may degenerate into pure ESS functions. Moreover, we found that the smaller the individual differences of RHP, the smaller is the mean pay-off of most individuals as well as the total pay-off of the population.Copyright 1998 Academic Press PMID- 9735264 TI - Modelling Coevolution in Multispecies Communities. AB - We introduce the Webworld model, which links together the ecological modelling of food web structure with the evolutionary modelling of speciation and extinction events. The model describes dynamics of ecological communities on an evolutionary time-scale. Species are defined as sets of characteristic features, and these features are used to determine interaction scores between species. A simple rule is used to transfer resources from the external environment through the food web to each of the species, and to determine mean population sizes. A time step in the model represents a speciation event. A new species is added with features similar to those of one of the existing species and a new food web structure is than calculated. The new species may (i) add stably to the web, (ii) become extinct immediately because it is poorly adapted, or (iii) cause one or more other species to become extinct due to competition for resources. We measure various properties of the model webs and compare these with data on real food webs. These properties include the proportions of basal, intermediate and top species, the number of links per species and the number of trophic levels. We also study the evolutionary dynamics of the model ecosystem by following the fluctuations in the total number of species in the web. Extinction avalanches occur when novel organisms arise which are significantly better adapted than existing ones. We discuss these results in relation to the observed extinction events in the fossil record, and to the theory of self-organized criticality.Copyright 1998 Academic Press PMID- 9735265 TI - Numbering Self-replicating Polymers. PMID- 9735268 TI - Adaptive Walks by the Fittest among Finite Random Mutants on a Mt. Fuji-type Fitness Landscape. AB - Based on the theory of fitness distributions on a Mt. Fuji-type fitness landscape in a multivalued sequence space (Aita & Husimi, 1996 J. theor. Biol. 182, 469 485), we investigated the properties of adaptive walks on the ideal landscape in the case of a cloning-screening-type evolution experiment. We modeled that an adaptive walk is performed by repetition of the evolution cycle composed of the mutagenesis process generating random d-fold point mutants of population size N and the selection process looking for the fittest mutant among them. While an adaptive walk is described in a sequence space, we simplified the description as follows. We mapped the landscape in an x-y plane, where x and y represent a normalized Hamming distance from the global peak and a scaled fitness, respectively. An adaptive walk is described as a trajectory in the plane. The most certain step for a walker to move in a single evolution cycle is represented by a vector in the plane. Then, a walker moves along the streams in the vector field determined by d and N. The walker performs fast hill-climbing until a "trap line", which traverses the plane. Subsequently, the walker is likely to get trapped in an "apparent local optimum". To continue the walk, apparent local optima must be eliminated by resetting d and N larger. Therefore, for the fastest walk, the optimal schedule of the d-values (initially large d, then small d) is effective, although the economical walk with high cost-performance is different. If a real landscape is just of the Mt. Fuji-type, the walk with the highest cost performance will be performed by scanning site-directed optimization through all sites. However, in the case of the rough Mt. Fuji-type, which seems to be more realistic, the walking method we have examined will be effective for a walker to sidestep true local optima.Copyright 1998 Academic Press PMID- 9735267 TI - The diffusion of molecules in axonal plasma membranes: the sites of insertion of new membrane molecules and their distribution along the axon surface. AB - The neuronal cell surface consists of two domains, the somatodendritic and axonal plasma membranes. Each domain serves different functions, and has a different complement of membrane molecules. Since membrane molecules are able to diffuse in the plane of the plasma membrane lipid bilayer, with diffusion coefficients ranging from 10-8 cm 2 s-1 for lipids to 10-10 cm 2 s-1 for proteins, mechanisms must exist to prevent as intermixing of membrane molecules from each domain by diffusion. Presented here is a theoretical analysis of the distribution of axonal molecules in both growing and non-growing axons based on two models for the insertion of these molecules into the axonal membrane, namely insertion exclusively at the distal end of the axon, or insertion with equal probability all along the axon. In all cases, assuming that the molecules have a finite half life in the axonal membrane, compositional differences between the axonal and somatodendritic membranes can be obtained that are similar to those observed in other polarized cells, even in the absence of a physical barrier to prevent the intermixing of membrane molecules. Moreover, our analyses demonstrate that the diffusion of membrane molecules in the plane of the axonal lipid bilayer is a sufficiently slow process to preclude the possibility that membrane molecules are inserted into axonal membranes at a site remote from their final location, and then move to their final locations for diffusion. Thus, in long axons, for membrane molecules that are localized all along the length of the axon, mechanisms must exist for their insertion in the axonal membrane at sites all along the axon, and not just at the distal end. PMID- 9735269 TI - Effects of Predator-prey Body Size Ratios on the Stability of Food Chains. AB - The effects of predator-prey body size ratios on the resilience and probability of stability in linear Lotka-Volterra food chains have been analysed. The prey per capita interaction strengths of the model is assumed to be negatively correlated to the relative size difference between a predator and its prey. The relationship between prey interaction strength and predator-prey body size ratios is motivated by energetical arguments. Analytical results show that, given this assumption (on prey interaction strengths) and if average (relative) size differences between predators and their prey decrease with the trophic position of the consumer (as found in a large number of "real food webs") the probability of local stability in model food chains is increased (when compared to model chains with a constant predator-prey body size ratio). Numerical simulations show that in most cases, the effect on the probability of stability is accompanied by an increase in resilience. For example, as model food chain length is increased from two to three trophic levels in one simulation, the return time increases by more than two orders of magnitude with a constant predator-prey body mass ratio while chains longer than four are not feasible. With a decreasing predator-prey body mass ratio on the other hand, the return time does not increase as rapidly and feasible equilibria exist for longer chains. The relationship between resilience and food chain length is, in this model, affected by the relationship between the predator-prey body mass ratio and the trophic position of the predator, that is, how fast this ratio decreases with increasing trophic height. The effect of body mass on consumer mortality rates, and subsequently on the probability of stability and resilience is also analysed. Decreasing mortality rates with increasing body size does not change the results qualitatively, it only increases the probability that an equilibrium is feasible.Copyright 1998 Academic Press PMID- 9735270 TI - The optimal production of gametocytes by Plasmodium falciparum. AB - We use a simple model of the blood-stage infection dynamics of the malaria parasite Plasmodium falciparum to consider the adaptive significance of different rates of conversion from its pathogenic, asexual stages to its transmissible, sexual forms. We find that maximize transmissivity in single-strain infections are generally greater than the highest rates reported for in vitro cultures and are several times those for which the behavior of the model is consistent with clinical profiles of infection dynamics. When two strains that share a common immune agent coinfect a host through simultaneous inoculation or sequential superinfection, however, a strain with a lower, clinically-consistent value of the conversion rate inhibits the transmissivity of one with the higher value optimal for single-strain infection. Hence we suggest that "apparent" competition by way of a common immune response might be responsible for selection of the former. PMID- 9735271 TI - A stochastic model for the rapid emergence of specific vertebrate immunity incorporating horizontal transfer of systems enabling duplication and combinational diversification. AB - Recent molecular data indicate that the antigen-specific combinatorial immune response is restricted to jawed vertebrates where it is found in representatives of all class from cartilagenous fishes to mammals. Here, we analyse the relatively rapid emergence of the combinatorial system terms of three stochastic process, with the system reaching essentially full capacity in immunoglobulin recognition elements and diversification and recombination of gene segments in an evolutionary span of time of less than 20 million years. The mechanisms for inducibility were coopted from ancient and widely spread processes in phylogeny for regulation of cell division. The proposed process of formation entailed the evolution of unknown ancestral genes into those specifying bona fide immunoglobulin domains, and the generation of multiple copies of these via a series of events facilitated by horizontal transfer of site-specific recombinases and recombination signal sequences most probably from microbial and fungal sources. The second process is one of rapid "decay" (evolution) which occurred in about 10 million year under stringent selective conditions to generate proper conserved canonical sequences. The third process is that of the long term evolution of these characteristic immunoglobulin domains over the 450 million years since their emergence. As a first approximation the rates of these three processes were computed using first order differential equations. The rate of formation has a magnitude of 10-7 substitutions per site per year, and that of rapid modifications is 10-8 substitutions per site per year. The long term rate of immunoglobulin evolution is comparable to that of other moderately conserved proteins, (1-3) x 10-9 substitutions per site per year). This model is testable by searching for "footprints" of microbial and fungal DNA processing enzymes and recombination mechanisms. The hypothesis raises the general concept that horizontal transfer of genes facilitating rearrangement and duplication can catalyse major steps of macroevolution. PMID- 9735272 TI - Habitat Destruction and Competitive Coexistence in Spatially Explicit Models with Local Interactions. AB - We investigate the competition-colonization trade-off in a two-species competition model in various heterogeneous environments. The competitiveness of species is hierarchically ordered and the dynamics of the model are described by spatially local interactions of individuals. The size of the local neighborhood may depend on the species. The habitat consists of inhabitable and permanently destroyed sites. It was previously observed (Nee & May, 1992; Tilman et al., 1994, 1997) that destruction of habitat may aid the inferior species and cause the superior species to go extinct. These conclusions were based on an n-species model in which the spatial arrangement of destroyed sites was ignored. We examine the effects of different spatial arrangements of destroyed sites on survival and coexistence of the two species. We conclude that the spatial arrangement plays an important role and can influence the outcome qualitatively. The key quantity for predicting the outcome of habitat destruction on species survival is the relationship between the mean dispersal distance and the mean distance between inhabitable and destroyed sites. We contrast the hierarchical model with a version of the Lotka-Volterra model of interspecific competition and show that habitat destruction can alter the dominance relationship between species by reducing intraspecific competition.Copyright 1998 Academic Press PMID- 9735273 TI - Sexual Reproduction and Stable Coexistence of Identical Competitors. AB - Current competition theory predicts that species must be ecologically distinct in order to stably coexist in the same community. This prediction is based on the implicit assumption that the consequences of sexual reproduction can be ignored. If anything, it is generally assumed that sex will only add problems, such as failure of insemination and inbreeding depression to species that have become rare, thus hastening competitive exclusion. Here we suggest that sexual reproduction may also bring rare species advantages in terms of the rate of per capita population growth. The key to our argument is that species-specific density dependence in population growth can occur when sexual reproduction is explicitly considered. We show that density-dependent sex ratio, sexual conflict, and sexually transmitted diseases can all promote species coexistence without separate niches.Copyright 1998 Academic Press PMID- 9735274 TI - A Resonance Model Gives the Response to Membrane Potential for an Ion Channel. AB - The current-voltage curve for ion channels is perhaps the best known characteristic of these channels. One of the first properties measured, it is accurately known for a variety of channels. The curve is usually described by a single thermal activation energy, which is assumed to show the number of channels opening in response to a voltage step. Activation allows movement of charges as the membrane depolarizes; the putative number of charges moving to open the channel is a parameter estimated from the curve. As the activation energy, E, creates a probability dependent on exp(-E/kBT) (kBT=Boltzmann's constantxtemperature) for a given channel to reach the open state, the opening probability distribution is referred to as a Boltzmann curve. The Boltzmann calculation of the complete curve is consistent with the experimental results on the i-V curve. However, other experimental data are not so easily explained by a Boltzmann curve, and there exists an alternative. A calculation based on the assumption of a threshold potential, which, if passed, allows a channel to open, leads to an open probability vs. potential curve which is also consistent with the measured current vs. voltage curve over its entire voltage range. The calculation assumes fluctuations in the local environment, leading to a distribution of the potential at which channels cross the threshold. There is a physical mechanism which would account for such a threshold, tunneling of a proton as the mechanism of charge movement. Actually, two or more almost independent tunneling transitions are required to obtain agreement with experiment, but this changes no essential feature of the model. Because tunneling requires matching of energy levels in two wells, it is appropriately referred to as a resonance. This model also makes it possible to explain an additional experiment: Fohlmeister & Adelman (1985a, b, 1986) have shown that a sinusoidal potential added to the membrane potential produces second harmonics of the sinusoidal frequency in the output current. Similar response to a sinusoidal input is found from the model, and compared qualitatively to the published experimental results of Fohlmeister & Adelman. A time delay can also be introduced between reaching the threshold and channel opening, which is both physically necessary and necessary for agreement with experiment. Unlike the Boltzmann model, out model agrees (qualitatively) with their experiment. We have tested two distributions of state as a function of potential: Gaussian, and skewed Gaussian ( reverse similarV2 exp(-aV2), V=potential). The latter comes closer to representing both the open probability-potential curve and the Fohlmeister & Adelman results, although the pure Gaussian is still better than the Boltzmann model.Copyright 1998 Academic Press PMID- 9735275 TI - The Coexistence of Competing Parasites. Part II-Hyperparasitism and Food Chain Dynamics. AB - Hyperparasitism is a widespread interaction in natural communities, but has to date received little attention in the theoretical literature. In this paper, we compared canonical models for food chains (resource-prey-predator systems) and host-parasite-hyperparasite interactions. We focus on microparasites, so the dynamical variables are the abundances of host individuals in different classes (e.g. with or without a particular parasite), and assume that the parasite is the only factor regulating a host population. Analysis of a "donor-controlled" model in which the primary parasite regulates host population growth, but with no additional demographic impact of the hyperparasite, suggests that intrinsic growth rate r of the host population is a fundamental parameter governing persistence of the hyperparasite. We then examine a model in which the hyperparasite can affect host births, deaths, and rate of recovery from the primary parasite. A wide range of outcomes are possible. For instance, hyperparasites can stabilize inherently unstable host-parasite systems, or destabilize stable systems. Persistence at a stable equilibrium often requires that the host intrinsic growth rate r lie within defined bounds; at low r, the hyperparasite may not be able to persist (in stable systems), whereas at high r the system is unstable and the host population grows in an unbounded fashion. We conclude by sketching directions for future work, and suggesting some possible practical implications of our results.Copyright 1998 Academic Press PMID- 9735276 TI - Developmental stability and signalling among cells. AB - The production of stable phenotypes depends from the earliest stages of development upon high levels of somatic cellular selection amongst cells or cell lineages. The signals exchanged amongst cells can reveal important aspects of a cell's phenotype, and might thereby be used in darwinian processes of developmental selection. Based upon an optimality model, we suggest that stable phenotypes require a substantial investment in two mechanisms of inter-cellular selection: "quality-selection" mechanisms regulate the average phenotype of a group of cells; "stability-selection" mechanisms regulate the variance in cell phenotypes. Variance in cell phenotypes may arise from developmental-error or other stochastic processes, or be generated as is true of the immune system, as part of a developmental strategy. The model shows that stability-selection mechanisms may exert the stronger effect on overall organ or body performance. Selection based upon reliable inter-cellular signalling of phenotypic properties may be the key way that bodies anticipate and then constrain variance in cell phenotypes around the optimal cellular attributes, and suggests an advantage of developmentally-selected systems over instructional ones. High levels of investments in stability mechanisms also ensure homogeneous collections of cells that can translate "upwards" into developmentally stable organ systems and phenotypes. Environmental and genetic factors, as well as the prevalent mode of selection, may all affect developmental stability and thereby give rise to varying of somatic selection. PMID- 9735277 TI - The derivation of knee joint types from the geometry of the cruciate ligament four-bar system. AB - The system of the anterior (a) and posterior (p) cruciate ligaments and their distances between attachments to femur (f) and tibia (t) as found in the knee joint of tetrapods is considered as a planar crossed four-bar linkage. The shape of the femoral articulating surfaces (condyles) can be calculated starting from a flat or curved tibial articulating surface and known bar-lengths (Menschik, 1974 Z. Orthop. 112, 481-495; Huson 1974 Orthopade 3, 119-126). Regression analysis of the dimensions of the cruciate ligament four-bar system of 11 species of mammal and one species of bird revealed a general ratio of (a): (t): (p): (f) = (7.1): (7.9): (10.0): (6.1). These data differ from the results obtained by Badoux (1984 Acta Anat. 119, 60-64) who examined only dog and horse. Our data of the dog agree with those of Badoux, i.e. (a): (t): (p): (f) approximately equal to (10): (8): (10): (4). Based on these ratios between bar-lengths, two types of knee joint shapes were distinguished. The shape of the dog's joint ("type A") has a very large femoral condyle compared with the tibial articulating surface. Maximum knee angulation is 170-180 degrees. Sliding between the articulating surfaces of this joint is distributed approximately uniformly over the whole angulation range. The general shape obtained from the regression analysis ("type R") has a relatively small femoral condyle and an angulation range of about 174 degrees. Uniformly distributed sliding occurs with this range over an angle less than 90 degrees. Theoretically derived, limiting requirements concerning maximum angulation range (delta gamma max < or = 180 degrees), stabilization (e.g. avoidance of a perpendicular position of the cruciate ligaments to the articulating surfaces; delta gamma 78.5 max > or = 90 degrees) and uniformly distributed sliding (delta gamma s > or = 30 degrees) lead to at least two different possible knee joint shapes. These shapes correspond to the two real knee joint shapes found from the statistical analysis mentioned above. This was verified by studying quantitative characteristics obtained from the derivation of knee joint shapes from the bar lengths and vice versa. The bird (Ardea) possessed a knee joint shape, very different from the shapes described above (i.e. f > t, type D1). PMID- 9735278 TI - Natural and induced tolerance in an immune network model. AB - It has been proposed that the immune system can be partitioned into central and peripheral immune systems. Recently, Carneiro et al. (1996a, b) proposed a network, model incorporating B and T lymphocytes that explicitly accounts for that partition. This model however, had some limitations that are tackled here. Two main changes were introduced: the average idiotypic connectivity is now an explicit function of time based on empirical evidence; and the activation of T lymphocytes by antigen is described by a log-bell shaped dose response curve. The new model, which also accounts for the CIS and PIS distinction, shows more reasonable results since the frequencies of tolerant, immune or autoimmune responses to an antigen are now correct. The model provides a new interpretation for tolerance induction during the neonatal period, and for the adult tolerance by low or high doses of antigen. It predicts that natural tolerance for antigens available during the neonatal period can be kept indefinitely upon their removal, while tolerance induced in the adult stages is rapidly lost upon transient removal of the antigen. A semiquantitative analysis of the model provides a simple explanation for the different results in terms of the frequency at which a limited set of canonical connectivity structures emerge during ontogenesis. PMID- 9735280 TI - Chaos, Dispersal and Extinction in Coupled Ecosystems. PMID- 9735279 TI - The Origin of Metazoa and the Egg: a Role for Cell Death. PMID- 9735281 TI - Two-dimensional crystallization and projection structure of KcsA potassium channel. AB - Potassium channels are integral membrane proteins that play a crucial role in regulating diverse cell functions in both electrically excitable and non excitable cells. Molecular cloning has revealed a diverse family of genes that encode these proteins, and a variety of experimental strategies have defined functional domains. We have cloned, over-expressed and purified the KcsA potassium channel to homogeneity and reconstituted this channel protein with phospholipids to form two-dimensional crystals. The crystals belong to plane group p4 and have unit cell dimensions of a=b=48 A. A projection map at 6 A resolution has been obtained by electron crystallography. The map shows that the protein is a homotetramer, having a low-density region on the 4-fold axis that is the site of the ion conduction pathway. Each monomer contains density features that are consistent with the molecular model of a truncated form of KcsA recently determined by X-ray crystallography. PMID- 9735282 TI - Computational determination of the structure of rat Fc bound to the neonatal Fc receptor. AB - The available crystal structure for the complex between the Fc fragment of immunoglobulin G (IgG) and the neonatal Fc receptor (FcRn) was determined at low resolution and has no electron density for a large portion of the CH2 domain of the Fc. Here, we use a well validated computational docking algorithm in conjunction with known crystallographic data to predict the orientation of CH2 when bound to FcRn, and validate the predicted structure with data from site specific mutagenesis experiments. The predicted Fc structure indicates that the CH2 domain moves upon binding FcRn , such that the end-to-end distance of the bound Fc fragment is greater than it is in the crystal structure of isolated Fc. The calculated orientation of the bound CH2 domain is displaced by an average of 6 A from the CH2 orientation in the structure of Fc alone, and shows improved charge complementarity with FcRn. The predicted effects of 11 specific mutations in Fc and FcRn are calculated and the results are compared with experimental measurements. The predicted structure is consistent with all reported mutagenesis data, some of which are explicable only on the basis of our model. The current study predicts that FcRn-bound Fc is asymmetric due to reorientation of the CH2 domain upon FcRn binding, a rearrangement that would be likely to interfere with optimal binding of FcRn at the second binding site of the Fc homodimer. PMID- 9735284 TI - In the absence of translation, RNase E can bypass 5' mRNA stabilizers in Escherichia coli. AB - In Bacilli, ribosomes or 30 S ribosomal subunits that are stalled or bound on mRNAs can stabilize downstream regions, hence the view that the degradation machinery scans mRNAs from their 5' end. In E. coli, several mRNAs can also be stabilized by secondary structures involving their 5' end. To test whether a bound 30 S subunit can act as a 5' stabilizer in E. coli, we compare here the stabilities of two untranslated variants of the lacZ mRNA, the decay of which is controlled by RNase E. In the first variant, a 35 nt region including the Ribosome Binding Site (RBS) is deleted, whereas in the second it is replaced by an 11 nt-long Shine-Dalgarno (SD) sequence lacking an associated start codon. In the latter variant, an 80 nt fragment encompassing the SD and extending up to the mRNA 5' end was stable in vivo (t1/2>one hour), reflecting 30 S binding. Yet, the full-length message was not more stable than when the SD was absent, although two small decay intermediates retaining the 5' end appear somewhat stabilized. A third variant was constructed in which the RBS is replaced by an insert which can fold back onto the lac leader, creating a putative hairpin involving the mRNA 5' end. The fragment corresponding to this hairpin was stable but, again, the full length message was not stabilized. Thus, the untranslated lacZ mRNA cannot be protected against RNase E by 5' stabilizers, suggesting that mRNA scanning is not an obligate feature of RNase E-controlled degradation. Altogether, these results suggest important differences in mRNA degradation between E. coli and B. subtilis. In addition, we show that mRNA regions involved in stable hairpins or Shine-Dalgarno pairings can be metabolically stable in E. coli. PMID- 9735283 TI - Nascent RNA in transcription complexes interacts with CspE, a small protein in E. coli implicated in chromatin condensation. AB - Proteins in a partially fractionated Escherichia coli extract that interact with the nascent RNA in active transcription complexes from several promoters were detected using the photocrosslinking ribonucleotide analogs 5-(azidophenacyl)thio UTP or 5-(azidophenacyl)thio-CTP as transcription substrates. Upon irradiation of ternary transcription complexes, several extract proteins were crosslinked to the RNA. Most notably, a small protein was crosslinked to the RNA in complexes on seven of nine templates tested. This protein was purified and sequenced and found to match a hypothetical protein, MsmC/CspE, recently shown to be involved in chromatin partitioning. CspE has 69% amino acid sequence identity with the major cold shock protein in E. coli, CspA, which has been shown to bind to a DNA sequence designated the Y box, with the sequence 5'-ATTGG. Of the nine templates tested, CspE was found to be most heavily crosslinked to RNA from the lambda PR' promoter, which is modified by the Q antiterminator protein. CspE was very heavily crosslinked to RNA only ten nucleotides long in initial ternary complexes on this promoter, but not to this same RNA after it had been released from the transcription complex. However, even when present from the start of transcription, CspE did not crosslink to the RNA 82 nucleotides long in elongation complexes from this same promoter. Despite the loss of interaction with the RNA after polymerase had left the promoter, CspE inhibited Q-mediated transcriptional antitermination from PR' in vitro almost 200 nucleotides downstream from the promoter, presumably by interaction with the Y box DNA upstream from PR', which overlaps with the binding site for the Q. A potential role for CspE and transcription in chromosome condensation and nucleoid structure is discussed. PMID- 9735285 TI - ErmE methyltransferase recognition elements in RNA substrates. AB - Dimethylation by Erm methyltransferases at the N-6 position of adenine 2058 (A2058, Escherichia coli numbering) in domain V of bacterial 23 S rRNA confers resistance to the macrolide-lincosamide-streptogramin B (MLS) group of antibiotics. The ErmE methyltransferase from Saccharopolyspora erythraea methylates a 625 nucleotide transcript of domain V as efficiently as it methylates intact 23 S rRNA. By progressively truncating domain V, the motif required for specific recognition by the enzyme has been localized to a helix and single-stranded region adjacent to A2058. The smallest RNA transcript that shows methyl-accepting activity is a 27-nucleotide stem-loop, corresponding to the 23 S rRNA sequences 2048 to 2063 and 2610 to 2620 (helix 73), with A2058 situated within the hairpin loop. Methylation of A2058 in the truncated RNAs is optimal in the absence of magnesium, and the efficiency of methylation is halved by the presence of 2 to 3 mM magnesium. Magnesium serves to stabilize a conformation in the truncated RNA that prevents efficient methylation. This contrasts to the intact domain V RNA, where 2 mM magnesium ions support a conformation at A2058 that is most readily recognized by ErmE. Methylation of domain V RNA is generally far less susceptible to ionic conditions than the truncated RNAs. The effects of monovalent cations on the methylation of truncated transcripts suggest that RNA structures outside helix 73 support the ErmE interaction. However, interaction with these structures is not essential for specific ErmE recognition of A2058. PMID- 9735286 TI - Mutations in domain III alpha of the Mu transposase: evidence suggesting an active site component which interacts with the Mu-host junction. AB - A series of point mutations was constructed in domain IIIalpha of the Mu A protein. The mutant transposases were purified and assayed for their ability to promote various aspects of the in vitro Mu DNA strand transfer reaction. All mutants with discernable phenotypes were inhibited in stable synapsis (Type 0 or Type 1 complex formation). In contrast, these mutant proteins were capable of LER formation (a transient early reaction intermediate in which the Mu left and right ends have been synapsed with the enhancer), at levels comparable to wild-type transposase. These proteins therefore comprise a novel class of transposase mutants, which are specifically inhibited in stable transpososome assembly. The defect in these proteins was also uniformly suppressed by either Mn2+, or the Mu B protein in the presence of ATP and target DNA. Striking phenotypic similarities were recognized between the domain IIIalpha transposase mutant characteristics noted above, and those for substrate mutants carrying a terminal base-pair substitution at the point of cleavage on the donor molecule. This phenotypic congruence suggests that the alterations in either protein or DNA are exerting an effect on the same step of the reaction i.e., engagement of the terminal nucleotide by the active site. We suggest that domain IIIalpha of the transposase comprises the substrate binding pocket of the active site which interacts with the Mu-host junction. PMID- 9735287 TI - SSB protein controls RecBCD enzyme nuclease activity during unwinding: a new role for looped intermediates. AB - The RecBCD enzyme of Escherichia coli initiates homologous recombination by unwinding and simultaneously degrading DNA from a double-stranded DNA end. Single stranded DNA loops are intermediates of this unwinding process. Here we show that SSB protein reduces the level of DNA degradation by RecBCD enzyme during unwinding, by binding to these ssDNA intermediates. Prior to interaction with the recombination hot spot chi, RecBCD enzyme has both 3'-->5' exonuclease and a weaker 5'-->3' exonuclease activity. We show that degradation of the 5'-terminal strand at the entry site is much more extensive in the absence of SSB protein. After interaction with chi, the level of 5'-->3' exonuclease activity is increased; as expected, degradation of the 5'-strand is also elevated in the absence of SSB protein. Furthermore, we show that, in the absence of SSB protein, the RecBCD enzyme is inhibited by the ssDNA products of unwinding; SSB protein alleviates this inhibition. These results provide insight into the organization of helicase and nuclease domains within the RecBCD enzyme, and also suggest a new level at which the nuclease activity of RecBCD enzyme is controlled. Hence, they offer new insight into the role of SSB protein in the initiation phase of recombination. PMID- 9735288 TI - The glycoinositolphospholipids from Leishmania panamensis contain unusual glycan and lipid moieties. AB - The cell surface of Leishmania parasites is coated by glycosylphosphatidylinositol (GPI)-anchored macromolecules (glycoproteins and a lipophosphoglycan) and a polymorphic family of free GPI glycolipids or glycoinositolphospholipids (GIPLs). Here we show that GIPLs with unusual glycan and lipid moieties are likely to be major cell surface components of L. panamensis (subgenus Viannia) promastigotes. These glycolipids were purified by high performance thin layer chromatography and their structures determined by gas liquid chromatography-mass spectrometry, fast-atom bombardment mass spectrometry, methylation analysis and chemical and enzymatic sequencing of the glycan headgroups. The major GIPLs contained two glycan core sequences, Manalpha1 3Manalpha1-4GlcN-phosphatidylinositol (type-2 series) or Manalpha1-3[Manalpha1 2Manalpha1-6]Manalpha1- 4GlcN-phosphatidylinosit ol (hybrid series), which were elaborated with Galalpha1-2Galbeta1- or Galalpha1-2/3Galalpha1-2Galbeta1- extensions that were attached to the 3-position of the alpha1-3 linked mannose. The phosphatidylinositol moiety contained exclusively diacylglycerol with palmitoyl, stearoyl and heptadecanoyl chains. Non-galactosylated GIPL species with the same core structures were also found. The galactose extensions and the presence of diacylglycerol in the lipid moieties are novel features for the GIPLs of Leishmania spp. The implications of these structures for the biosynthesis of leishmanial GIPLs and their putative function in the mammalian host are discussed. PMID- 9735289 TI - Mapping the mAb 383C epitope to alpha 2(187-199) of the Torpedo acetylcholine receptor on the three-dimensional model. AB - Monoclonal antibody 383C is an anti-acetylcholine receptor antibody whose binding to the receptor is blocked by alpha-bungarotoxin and by carbamylcholine. Monoclonal antibody 383C binds to the alpha subunit of the Torpedo acetylcholine (ACh) receptor as well as to its V8-protease 20 kDa fragment that possesses the affinity alkylatable Cys192/193. In an epitope scanning experiment spanning the N terminal 211 amino acid residues of the alpha subunit, 383C binds uniquely to three overlapping peptides; alpha(184-196), alpha(187-199) and alpha(190-202). These peptides span a cluster of amino acid residues implicated in the binding of acetylcholine, including Cys192/193. To map the location of these residues on the three-dimensional model of the ACh receptor, we have employed a combination of X ray diffraction from oriented complexes of 383C with ACh receptor-enriched membrane vesicles and electron microscopy of negatively stained tubular arrays of 383C/receptor complexes. The X-ray diffraction study finds extra electron density in the presence of 383C centered 35 A above the synaptic side phosphate head groups. The electron micrographic images display extra stain exclusion from the antibody at a site adjacent to the alpha2 subunit on the periphery of the rosette clockwise to the alpha2 vertex. This mapping localizes several residues of the ACh receptor alpha subunit involved in the binding of acetylcholine. Despite these residues being present in both alpha subunits, only the alpha2 subunit is decorated with this monoclonal antibody. PMID- 9735291 TI - Two distinct modes of protein-induced bending in DNA. AB - Crystallised "naked" DNA oligomers in the B form show significant conformational mobility, particularly at CA/TG and TA/TA steps: there is a range in Roll angle of some 15 degrees between consecutive base-pairs, and Slide and Twist are directly coupled to Roll. We call such motions "mode I". They are sufficient to enable DNA to curve gently around proteins such as histone octamers in the nucleosome particle. When DNA bends around other proteins, such as CAP and TBP, its distortion is much more severe. Although the DNA in close contact with these proteins includes the CA/TG and TA/TA steps, respectively, the mode I flexibility is not deployed: instead, a more severe "mode II" manoeuvre is observed in DNA/protein co-crystals. Mode II has several distinctive physical features. First, its range of Roll angle is much wider than for mode I. Second, the major groove width remains more-or-less constant as Roll increases, whereas it decreases significantly as Roll increases in mode I; and this enables the major groove of the DNA to accommodate a protein moiety in its severely bent conformation. Third, the value of Slide remains more-or-less constant as Roll increases, whereas it decreases in mode I. In general, in both modes I and II, the major-groove width appears to be closely related to the Slide between base pairs. In mode II there appears to be a definite "point pivot" on the major groove side of the two base-pairs that constitute a dinucleotide step, formed either by the steric interlocking of propeller-twisted base-pairs or by a bifurcated hydrogen bond. Distortion of DNA in mode II seems to be an intrinsic property of the double-helical structure, since it occurs whether protein is bound on the major-groove side (e.g. CAP) or on the minor-groove side (e.g. TBP). Mode II distortion occurs in a wider range of steps than those that show the largest mode-I variation; nevertheless, "access" to mode II deformation appears to be gained via mode I distortion at particular steps CA/TG and TA/TA. PMID- 9735290 TI - Differential surface accessibility of alpha(187-199) in the Torpedo acetylcholine receptor alpha subunits. AB - We have probed the surface accessibility of residues alpha187 to alpha199 of the Torpedo acetylcholine receptor with monoclonal antibody 383C, which binds uniquely to these residues. However, 383C binds to only one of the two alpha subunits in the membrane-bound receptor, neither of the two subunits in carbamylcholine-desensitized receptor, and to both alpha subunits in Triton X-100 solubilized receptor. The kinetics of association and dissoci-ation of 383C with the peptide alpha(183-199) compared to those with the membrane-bound receptor suggest that all but a single hydrogen bond of affinity derives from contacts between this peptide and the monoclonal antibody paratope. Inhibition of 383C binding by alpha-bungarotoxin selectively directed to the alpha subunit correlated with the high-affinity d-tubocurarine binding site, along with a lack of inhibition by alpha-bungarotoxin directed to the alpha subunit correlated with the low-affinity d-tubocurarine binding site, suggests that the 383C epitope on the membrane-bound receptor resides on the alpha subunit associated with the high affinity d-tubocurarine binding site. The results presented here suggest a structural basis for the differences between the two receptor acetylcholine binding sites. PMID- 9735292 TI - Regulation of hexokinase I: crystal structure of recombinant human brain hexokinase complexed with glucose and phosphate. AB - Hexokinase I, the pacemaker of glycolysis in brain tissue and red blood cells, is comprised of two similar domains fused into a single polypeptide chain. The C terminal half of hexokinase I is catalytically active, whereas the N-terminal half is necessary for the relief of product inhibition by phosphate. A crystalline complex of recombinant human hexokinase I with glucose and phosphate (2.8 A resolution) reveals a single binding site for phosphate and glucose at the N-terminal half of the enzyme. Glucose and phosphate stabilize the N-terminal half in a closed conformation. Unexpectedly, glucose binds weakly to the C terminal half of the enzyme and does not by itself stabilize a closed conformation. Evidently a stable, closed C-terminal half requires either ATP or glucose 6-phosphate along with glucose. The crystal structure here, in conjunction with other studies in crystallography and directed mutation, puts the phosphate regulatory site at the N-terminal half, the site of potent product inhibition at the C-terminal half, and a secondary site for the weak interaction of glucose 6-phosphate at the N-terminal half of the enzyme. The relevance of crystal structures of hexokinase I to the properties of monomeric hexokinase I and oligomers of hexokinase I bound to the surface of mitochondria is discussed. PMID- 9735293 TI - Crystal structures of the catalytic domain of HIV-1 integrase free and complexed with its metal cofactor: high level of similarity of the active site with other viral integrases. AB - Human immunodeficiency virus (HIV) integrase is the enzyme responsible for insertion of a DNA copy of the viral genome into host DNA, an essential step in the replication cycle of HIV. HIV-1 integrase comprises three functional and structural domains: an N-terminal zinc-binding domain, a catalytic core domain and a C-terminal DNA-binding domain. The catalytic core domain with the F185H mutation has been crystallized without sodium cacodylate in a new crystal form, free and complexed with the catalytic metal Mg2+. The structures have been determined and refined to about 2.2 A. Unlike the previously reported structures, the three active-site carboxylate residues (D,D-35-E motif) are well ordered and both aspartate residues delineate a proper metal-binding site. Comparison of the active binding site of this domain with that of other members from the polynucleotidyl transferases superfamily shows a high level of similarity, providing a confident template for the design of antiviral agents. PMID- 9735294 TI - X-ray structure of a blue-copper nitrite reductase in two crystal forms. The nature of the copper sites, mode of substrate binding and recognition by redox partner. AB - Denitrification is one of the main steps of the global nitrogen cycle that is sustained by prokaryotic organisms. Denitrifying bacteria use two entirely different enzymes in this process, one based on haem cd1 prosthetic groups and the other on type 1-type 2 Cu centres. Copper-containing nitrite reductases (NiRs) are sub-divided into blue and green NiRs, which are respectively thought to be redox partners of azurins and pseudo-azurins. Crystallographic structures of the blue nitrite reductase from Alcaligenes xylosoxidans (AxNiR) are presented in the oxidised hexagonal form and the substrate-bound orthorhombic form to 2.1 A and 2.8 A resolution, respectively. The complete amino acid sequence of AxNiR has been determined by conventional chemical analysis. A 3 A structure of AxNiR has been published where the modelling was based on the sequence of another blue NiR. The higher resolution of the hexagonal form together with the correct sequence allows a detailed comparison with the crystallographic structures of the green NiRs. There is a striking difference in the overall surface charge distribution between the two sub-groups, providing a neat structural explanation for their different reactivities to pseudoazurin or azurin and supporting the view that electron transfer proceeds via complex formation. A detailed examination of the type-1 Cu site, the site responsible for the colour, reveals several subtle differences, including a lateral displacement of 0.7 A for Smet. The structure of the type-2 Cu site, and changes that occur upon substrate binding are discussed in terms of the catalytic mechanism. The similarity of the type 2 Cu site to the catalytic Zn site in carbonic anhydrase and the catalytic Cu site of superoxide dismutase is re-examined in view of the high-resolution (2.1 A) structure. PMID- 9735295 TI - The refined crystal structure of Drosophila lebanonensis alcohol dehydrogenase at 1.9 A resolution. AB - Drosophila alcohol dehydrogenase (DADH; EC 1.1.1.1) is a NAD(H)-dependent oxidoreductase belonging to the short-chain dehydrogenases/reductases (SDR) family. This homodimeric enzyme catalyzes the dehydrogenation of alcohols to their respective ketones or aldehydes in the fruit-fly Drosophila, both for metabolic assimilation and detoxification purposes. The crystal structure of the apo form of DADH, one of the first biochemically characterized member of the SDR family, was solved at 1.9 A resolution by Patterson methods. The initial model was improved by crystallographic refinement accompanied by electron density averaging, R-factor=20.5%, R-free=23.8%.DADH subunits show an alpha/beta single domain structure with a characteristic NAD(H) binding motif (Rossmann fold). The peptide chain of a subunit is folded into a central eight-stranded beta-sheet flanked on each side by three alpha-helices. The dimers have local 2-fold symmetry. Dimer association is dominated by a four-helix bundle motif as well as two C-terminal loops from each subunit, which represent a unique structural feature in SDR enzymes with known structure. Three structural features are characteristic for the active site architecture. (1) A deep cavity which is covered by a flexible loop (33 residues) and the C-terminal tail (11 residues) from the neighboring subunit. The hydrophobic surface of the cavity is likely to increase the specificity of this enzyme towards secondary aliphatic alcohols. (2) The residues of the catalytic triad (Ser138, Tyr151, Lys155) are known to be involved in enzymatic catalysis in the first line. The Tyr151 OH group is involved in an ionic bond with the Lys155 side-chain. Preliminary electrostatic calculations have provided evidence that the active form of Tyr151 is a tyrosinate ion at physiological pH. (3) Three well-ordered water molecules in hydrogen bond distance to side-chains of the catalytic triad may be significant for the proton release steps in DADH catalysis.A ternary structure-based sequence alignment with ten members of the SDR family with known three-dimensional structure has suggested to define a model consisting of four groups of residues, which relates the observed low degree of sequence identity to quite similar folding patterns and nearly identical distributions of residues involved in catalysis. PMID- 9735297 TI - NMR structure of the Streptomyces metalloproteinase inhibitor, SMPI, isolated from Streptomyces nigrescens TK-23: another example of an ancestral beta gamma crystallin precursor structure. AB - The Streptomyces metalloproteinase inhibitor, SMPI, isolated from Streptomyces nigrescens TK-23, is a proteinaceous metalloproteinase inhibitor, and consists of 102 amino acid residues with two disulfide bridges. SMPI specifically inhibits metalloproteinases such as thermolysin. In the present work, the solution structure of SMPI was determined on the basis of 1536 nuclear Overhauser enhancement derived distance restraints and 52 dihedral angle restraints obtained from three-bond spin coupling constants. The final ensemble of 20 NMR structures overlaid onto their mean coordinate with backbone (N, Calpha, C') r.m.s.d. values of 0. 45(+/-0.11) A and 0.57(+/-0.18) A for residues 6 to 99 and the entire 102 residues, respectively. SMPI is essentially composed of two beta-sheets, each consisting of four antiparallel beta-strands. The structure can be considered as two Greek key motifs with 2-fold internal symmetry, a Greek key beta-barrel. One unique structural feature found in SMPI is in its extension between the first and second strands of the second Greek key motif. Interestingly, this extended segment is known to be involved in the inhibitory activity of SMPI. In the absence of sequence similarity, the SMPI structure shows clear similarity to both domains of the eye lens crystallins, both domains of the calcium sensor protein S, as well as the single-domain yeast killer toxin. The yeast killer toxin structure was thought to be a precursor of the two-domain beta gamma-crystallin proteins, because of its structural similarity to each domain of the beta gamma crystallins. SMPI thus provides another example of a single-domain protein structure that corresponds to the ancestral fold from which the two-domain proteins in the beta gamma-crystallin superfamily are believed to have evolved. PMID- 9735296 TI - Solution structure of the M13 major coat protein in detergent micelles: a basis for a model of phage assembly involving specific residues. AB - The three-dimensional structure of the major coat protein of bacteriophage M13, solubilized in detergent micelles, has been determined using heteronuclear multidimensional NMR and restrained molecular dynamics. The protein consists of two alpha-helices, running from residues 8 to 16 and 25 to 45, respectively. These two helices are connected by a flexible and distorted helical hinge region. The structural properties of the coat protein make it resemble a flail, in which the hydrophobic helix (residues 25 to 45) is the handle and the other, amphipathic, helix the swingle. In this metaphor, the hinge region is the connecting piece of leather. The mobility of the residues in the hinge region is likely to enable a smooth transformation from the membrane-bound form, mimicked by the structure in detergent micelles, into the structure in the mature phage. A specific distribution of the residues over the surface of the two helices was observed in the presented high-resolution structure of the membrane-bound form of the major coat protein as well as in the structure in the mature phage. All data suggest that this arrangement of residues is important for the interactions of the protein with the membrane, for correct protein-DNA and protein-protein interactions in the phage and for a proper growth of the phage during the assembly process. By combining our findings with earlier NMR results on the major coat protein in detergent micelles, we were able to construct a model that addresses the role of specific residues in the assembly process. PMID- 9735298 TI - Elucidation of the mode of interaction of thermolysin with a proteinaceous metalloproteinase inhibitor, SMPI, based on a model complex structure and a structural dynamics analysis. AB - SMPI is a proteinaceous microbial metalloproteinase inhibitor that was isolated from Streptomyces nigrescens TK-23 in 1979. SMPI is known to selectively inhibit the metalloproteinases in the gluzincin family, according to the Rawling and Barrett classification. There has been no report on the interaction of a metalloproteinase in the family of gluzincins with its specific proteinaceous inhibitor. We have solved the solution structure of SMPI by NMR. Here, we report the binding mode of SMPI to thermolysin, based on the model complex structure generated using our high-resolution NMR structure of SMPI and the crystal structure of thermolysin. The obtained complex model shows that the extruded loop of SMPI, with the scissile bond Cys64-Val65, is complementary in shape to the active cleft of thermolysin. In the complex, the Cys64 (P1) carbonyl oxygen atom can form a tetrahedral coordination to the active zinc in thermolysin, and simultaneously, the methyl groups of Val65 (P1') are closely located in the hydrophobic S1' pocket in thermolysin. From the electrostatic potential surface calculation, the active loop of SMPI and the active cleft in thermolysin have been shown to be complementary in the surface charge distribution, resulting in the stabilization of the complex. The apparently large active loop is less flexible, but maintains a conformation in the nano- to picosecond time-scale, as elucidated from the 15N spin relaxation analysis. This is a quite different structural feature of SMPI from the flexible binding loop generally found in the serine proteinase inhibitors, such as SSI and eglin c, and can be related to the narrow specificity of SMPI. The present study provides the first insight into the interaction between a proteinaceous inhibitor and a gluzincin metalloproteinase. PMID- 9735299 TI - The role of threonine in the P2 position of Bowman-Birk proteinase inhibitors: studies on P2 variation in cyclic peptides encompassing the reactive site loop. AB - Previously, we have described a template-assisted combinatorial peptide library based on the anti-tryptic reactive site loop of a Bowman-Birk inhibitor (BBI). Sequences that displayed inhibitory activity re-directed towards chymotrypsin were found to have a consensus binding motif, with their most striking feature being that exclusively threonine was found at the P2 position. The present study investigates the reason for this surprising specificity by maintaining the binding motif but systematically varying the P2 residue. From analysis of 26 variants, it is found that the requirements for inhibitory activity at P2 are finely tuned, and in agreement with the library work, threonine at P2 provides optimal inhibition. In addition, peptides with threonine at P2 are significantly less susceptible to hydrolysis. Examination of all available BBI sequences shows that threonine is very highly conserved at P2, which implies that the functional requirement extends to the full-length BBI protein. Our results are consistent with a dual requirement for hydrophobic recognition within the S2 pocket and maintenance of an inhibitory conformation via hydrogen bonding within the reactive-site loop. As the isolated peptide loop reproduces the active region of full-length BBI, these results explain why threonine is well conserved at P2 in this class of inhibitor. Furthermore, they illustrate that proteinase inhibitor specificity can have characteristics that are not easily predicted from information on the substrate preferences of a proteinase. PMID- 9735301 TI - Protein-nucleic acid interactions and cellular responses to interferon. PMID- 9735300 TI - Evolutionary conserved rigid module-domain interactions can be detected at the sequence level: the examples of complement and blood coagulation proteases. AB - Several extracellular modular proteins, including proteases of the complement and blood coagulation cascades, are shown here to exhibit conserved sequence patterns specific for a particular module-domain association. This was detected by comparative analysis of sequence variability in different multiple sequence alignments, which provides a new tool to investigate the evolution of modular proteins. A first example deals with the proteins featuring a common complement control protein (CCP) module-serine protease (SP) domain pattern at their C terminal end, defined here as the CCP-SP sub-family. These proteins include the complement proteases C1r, C1s and MASPs, the Limulus clotting factor C, and the proteins of the haptoglobin family. A second example deals with blood coagulation factors VII, IX and X and protein C, all featuring a common epidermal growth factor (EGF)-SP C-terminal assembly. Highly specific motifs are found at the connection between the CCP or EGF module and the activation peptide of the SP domain: [P/A]-x-C-x-[P/A]-[I/V]-C-G-x-[P/S/K] in the case of the CCP-SP proteins, and C-x-[P/S]-x-x-x-[Y/F]-P-C-G in the case of the EGF-SP proteins. Each motif is strictly conserved in the whole sub-family and it is detected in no more than one other known protein sequence. Strikingly, most of the conserved residues specific to each sub-family appear to be clustered at the interface between the SP domain and the CCP or EGF module. We propose that a rigid module-domain interaction occurs in these proteins and has been conserved through evolution. The functional implications of these assemblies, underlined by such evolutionary constraints, are discussed. PMID- 9735302 TI - Analysis of interferon-regulated proteins binding the interferon-alpha-stimulated response element. AB - Interferon mediates its biological effects of antiviral, antiproliferative, and immunomodulatory activities through induction of specific gene expression. Transcription of target genes is regulated through a set of transcription factors that bind to specific cis-acting regulatory sequences in target promoters and enhancers. The activity or abundance of these transcription factors is modulated by interferon treatment. Because of their DNA binding activity, regulatory factors can be recognized and characterized using methods to detect protein-DNA complexes. The availability of antibodies directed against common interferon regulatory proteins coupled with analysis of binding-site specificity provides comprehensive analysis. PMID- 9735303 TI - Protein-DNA interactions in interferon-gamma signaling. AB - The ability to rapidly activate new genes is essential for the biological effects mediated by IFN-gamma. Studies directed at understanding how these genes are induced by this ligand led to the identification of the STAT family of transcription factors. STATs are rapidly activated at the receptor, whereupon they translocate to the nucleus and bind to a unique enhancer found in the promoter of target genes. The ability to identify this IFN-gamma response element and the proteins that bind it was critical for the elucidation of this pathway. These techniques are the focus of this review. PMID- 9735304 TI - RNA binding and modulation of PKR activity. AB - PKR is an RNA-dependent protein kinase that is induced in mammalian cells by interferon treatment. It is present in a latent or inactive form in mammalian cells and is activated by very low concentrations of double-stranded (ds) RNA. Activated PKR phosphorylates eIF2, an essential initiation factor of protein synthesis, as well as other substrates including histone IIA, a 90-kDa protein from rabbit reticulocytes, the inhibitor, IkappaB, of the transcription factor, NF-kappaB, and the HIV-1 Tat protein. PKR interacts with several cellular and viral products and these interactions modulate its activation by dsRNA. Here we describe methods that are used to study the activation or inhibition of PKR by RNA modulators. Specifically, we detail (1) the purification of PKR from interferon-treated mammalian cells, (2) functional assays for PKR activation and inhibition in vitro, using purified enzyme or crude cell lysates, and (3) assays allowing evaluation of the binding of dsRNA and single-stranded RNA to PKR. PMID- 9735305 TI - Double-stranded RNA-specific adenosine deaminase: nucleic acid binding properties. AB - The RNA-specific adenosine deaminase (ADAR1, herein referred to as ADAR) is an interferon-inducible RNA-editing enzyme. ADAR catalyzes the C-6 deamination of adenosine in double-stranded (ds) structures present in viral RNAs and cellular pre-mRNAs as well as synthetic dsRNA substrates. ADAR possesses three functionally distinct copies of the highly conserved double-stranded RNA binding R motif (RI, RII, RIII) implicated in the recognition of dsRNA structures within the substrate RNAs. ADAR is also a Z-DNA-binding protein. Two Z-DNA binding motifs (Zalpha and Zbeta) present in ADAR correspond to repeated regions homologous to the N-terminal region of the vaccinia virus E3L protein. Here we describe assay methods for measurement of ADAR enzymatic activity, dsRNA binding activity, and Z-DNA binding activity. PMID- 9735306 TI - Using genetic means to dissect homologous and heterologous protein-protein interactions of PKR, the interferon-induced protein kinase. AB - The interferon-induced protein kinase, PKR, is a pivotal component of interferon (IFN)-induced cellular antiviral and antiproliferative response. The identification and characterization of proteins, of both viral and cellular origins, that interact with PKR have proven to be a valuable probe for unraveling the cellular regulation and function of PKR. Several studies have demonstrated that PKR forms dimers and that dimerization is likely to be required for activation and/or catalytic function. It is therefore important to elucidate the mechanism of PKR dimer formation and the role of PKR effectors in modulating kinase dimerization. Herein we describe the use of the two genetic approaches, the lambda repressor fusion and the yeast two-hybrid systems, to detect and analyze homo- and heterotypic interactions with PKR. We also describe several biochemical methodologies commonly used in our laboratory to validate the genetic results. Although the examples in this article focus on PKR, the techniques can easily be adapted to investigate protein-protein associations in a variety of experimental systems. Finally, given the important role of PKR as a mediator of IFN-induced antiviral and antiproliferative effects, these studies may provide clues to the development of reagents that target PKR to enhance the therapeutic use of IFN in the treatment of disease. PMID- 9735307 TI - Characterization of viral double-stranded RNA-binding proteins. AB - Several viruses have been shown to code for proteins that specifically bind to double-stranded RNA or RNA with large amounts of secondary structure. These proteins have been implicated in providing interferon resistance to viruses and in inhibiting induction of apoptosis by viruses, and have been suggested to be involved in regulation of viral and cellular protein synthesis in infected cells. This article describes methods for detecting and analyzing viral double-stranded RNA-binding proteins. PMID- 9735308 TI - Production, purification, and characterization of recombinant 2', 5' oligoadenylate synthetases. AB - 2',5'-Oligoadenylate [2-5(A)] synthetases are a family of interferon-induced enzymes that polymerize ATP into 2'-5'-linked oligoadenylates in the presence of double-stranded RNA (dsRNA), their cofactor. The 2-5(A) molecules, in turn, activate the latent ribonuclease RNase L by promoting its dimerization. The 2 5(A) synthetase pathway has been implicated in interferon's antiviral and anticellular activities. In addition to their interesting cellular properties, these enzymes are also enzymologically interesting because they are the only known template and primer independent nucleotide (DNA or RNA)polymerases that synthesize 2'-5'-linked oligonucleotides. Moreover, their mode of activation by dsRNA remains unknown. In the past, biochemical and structure-function studies have been hampered by the lack of a convenient system for expressing recombinant 2-5(A) synthetases. These proteins are toxic to mammalian cells, probably because of RNase L activation, and proteins produced in bacteria do not have full enzymatic activity. To circumvent these problems, we have developed a baculovirus insect cell system for high-yield expression of the small and medium isozymes. Here, methods are described for the production, purification, and characterization of the mouse small (9-2) (S. K. Ghosh, J. Kusari, S. K. Bandyopadhyay, H. Samanta, R. Kumar, and G. C. Sen, 1991, J. Biol. Chem. 266, 15293-15299) and human medium (P69) (I. Marie and A. G. Hovanessian, 1992, J. Biol. Chem. 267, 9933-9939) 2-5(A) synthetase isozymes and their mutants using the insect cell system. We also report methods for studying 2-5(A) synthetase dsRNA interactions and protein-protein interactions among the subunits of the two isozymes. PMID- 9735309 TI - Ribonuclease L, a 2-5A-dependent enzyme: purification to homogeneity and assays for 2-5A binding and catalytic activity. AB - RNase L is a latent endonuclease found in reptiles, birds, and mammals. It is activated by the 2',5'-phosphodiester-linked oligoadenylates called 2-5A and has been implicated in the mechanism of action of interferon, as well as in a variety of other biological phenomena such as apoptosis. Covalent linkage of 2-5A to antisense oligonucleotides permits recruitment of RNase L for enhancement of antisense action. The purification of RNase L described herein and the assays for its detection and activation will help to provide further mechanistic details on how this unique nuclease functions and what its biochemical roles may be. In addition, such assays will facilitate the screening of 2-5A-antisense congeners for exploration of the potential therapeutic applications of RNase L. PMID- 9735310 TI - MxA GTPase: oligomerization and GTP-dependent interaction with viral RNP target structures. AB - MxA protein is an interferon-induced GTPase of human cells that inhibits the multiplication of several RNA viruses, including influenza viruses and bunyaviruses. Studies on MxA transgenic mice have shown that MxA is a powerful antiviral agent in vivo. It has been suggested that this cellular protein also protects humans from viral disease, but the mechanism(s) by which MxA exerts its antiviral action is still poorly understood. Using an in vitro cosedimentation assay, we now demonstrate that MxA tightly interacts with components of the ribonucleoprotein complex of Thogoto virus, an influenza-like virus transmitted by ticks. This assay demonstrates for the first time a physical interaction between MxA GTPase and a viral target structure. It is based on three elements, namely, highly active MxA GTPases as effector molecules, viral ribonucleoprotein particles as viral targets, and GTPgammaS as a stabilizing factor. Furthermore, using a simple nuclear translocation assay, we show that human MxA protein forms oligomers in vivo. This assay provides a stringent test for tight association of partner molecules in intact mammalian cells. It not only will be useful for studying physical interactions of MxA with partner molecules, but may also be applicable to other studies on protein-protein interactions in living cells. PMID- 9735311 TI - The central lysine in the P-loop motif of the Escherichia coli DnaA protein is essential for initiating DNA replication from the chromosomal origin, oriC, and the F factor origin, oriS, but is dispensable for initiation from the P1 plasmid origin, oriR. AB - The Escherichia coli DnaA protein is essential for initiation of DNA replication from the chromosomal origin, oriC, and from certain plasmid origins such as oriR of P1, oriS of F, and ori of pSCS101. The DnaA protein binds ATP with high affinity and contains a P-loop motif assumed to be the binding site. Three mutations in the E. coli dnaA gene were constructed by oligonucleotide-directed mutagenesis that changed amino acids in the P-loop. A DnaA protein, K178T, in which the central lysine was changed to the smaller amino acid threonine, was able to initiate DNA replication from P1 oriR, but was unable to initiate replication from E. coli oriC or F oriS in vivo. Mutant and wild-type DnaA proteins were overexpressed, partially purified, and tested for replication activity in vitro. The K178T DnaA protein could initiate replication from oriR, although with a decreased activity compared to the wild-type DnaA protein. No replication activity was detected for this mutant protein from oriC. The different responses of the oriR and oriC replicons to the K178T DnaA protein indicate that the role of DnaA is different in the two systems. PMID- 9735312 TI - Isolation and characterization of plasmid pSW200 from Erwinia stewartii. AB - The nucleotide sequence of pSW200 of Erwinia stewartii SW2 was determined. This plasmid is 4367 bp long, consisting of four mobilization genes, mobCABD, and an origin of replication homologous to those of ColE1-type plasmids. The plasmid also contains a region of forty-one 15-bp repeats. Deleting this region does not affect the stability or the copy number when maintained as sole plasmid in the cell. However, the plasmid is rapidly lost when a homoplasmid with the intact repeat region is introduced into the cell. The function of this region may provide pSW200 an advantage in competing with an incompatible plasmid in the cell. PMID- 9735313 TI - Replication and maintenance of lambda plasmids devoid of the Cro repressor autoregulatory loop in Escherichia coli. AB - Plasmids derived from bacteriophage lambda are known as lambda plasmids. These plasmids contain the ori lambda region and lambda replication genes O and P. Typical lambda plasmids also contain the cro gene, the product of which is a repressor of the pR promoter when present at relatively high concentrations. These genes stably maintain the plasmid in Escherichia coli at copy numbers of 20 to 50 per cell. According to a generally accepted model, stable maintenance of lambda plasmids is possible due to the Cro repressor autoregulatory loop (the cro gene is under control of pR). Here we demonstrate that lambda plasmids devoid of the Cro autoregulatory loop can also be stably maintained in E. coli strains. We present data for two such plasmids: pTC lambda 1 in which the pR-cro region has been replaced by the ptetA promoter and the tetR gene (coding for the TetR repressor), and a standard lambda plasmid with inactivated cro gene (lambda cro null plasmid). Thus, the presence of the Cro repressor autoregulatory loop does not appear to be essential to the maintenance of lambda plasmids in vivo. PMID- 9735314 TI - Corynebacterium striatum chloramphenicol resistance transposon Tn5564: genetic organization and transposition in Corynebacterium glutamicum. AB - The clinical isolate Corynebacterium striatum M82B (formerly Corynebacterium xerosis M82B) carries the 50-kb R-plasmid pTP10 conferring resistance to the antibiotics chloramphenicol, erythromycin, kanamycin, and tetracycline. DNA sequence analysis of the chloramphenicol resistance region revealed the presence of the 4155-bp transposable element Tn5564. The ends of Tn5564 are identical 22 bp inverted repeats flanked by a 6-bp target site duplication. The central region of Tn5564 encodes the chloramphenicol resistance gene cmx, specifying a transmembrane chloramphenicol efflux protein, and an open reading frame homologous to transposases of insertion sequences identified in Arthrobacter nicotinovorans and Bordetella pertussis. Furthermore, the 1715-bp insertion sequence IS1513 encoding a putative transposase of the IS30 family is an integral part of Tn5564 and is located upstream of cmx. For transposon mutagenesis, Tn5564 was transferred to Corynebacterium glutamicum on a mobilizable Escherichia coli plasmid using RP4-mediated intergeneric conjugation. Transposition of Tn5564 in C. glutamicum occurred with a frequency of 3.3 x 10(-8) and resulted in an insertion into target sites containing the central palindromic tetranucleotide CTAG. A Tn5564-induced mutant strain of C. glutamicum was found to carry the transposon in the ftsZ gene region. PMID- 9735315 TI - Functional domains of Rts1 and P1 RepA proteins for initiation of replication. AB - Rts1 RepA and P1 RepA are trans-acting proteins essential for initiation of replication of Rts1 and P1 plasmids, respectively. We recently found that P1 RepA bound in vitro to the Rts1 replication origin as strongly as Rts1 RepA and activated the origin in vivo. However, the ori activation was quite inefficient. This study shows that by replacing a small region of P1 RepA with the corresponding region of Rts1 RepA, the efficiency of Rts1 ori activation increased markedly. Interestingly, the same subregion of P1 RepA was found to be important for in vivo activation of the P1 origin. Thus, a region essential for efficient activation of the replication origin was assigned to the P1 RepA molecule as well as to the Rts1 RepA molecule. The region was distinct from a domain necessary for in vitro binding to the origin, although both regions were required for in vivo activation of the respective origin. PMID- 9735317 TI - Biological behavior of plasmid in Rhizobium sp. strain S25 from Tephrosia candida. AB - Rhizobium sp. strain S25 was isolated from the nodule on Tephrosia candida in Hainan Province, China. The strain showed high stress tolerance. The plasmid profile of strain S25, examined by the Eckhardt procedure, indicated that the strain harbors only one plasmid with an estimated size of 150 kb. The plasmid was shown to carry nod and nif genes by hybridization with probes of nodABC and nifHDK genes. Plasmid curing was carried out using the Bacillus subtilis sacB to generate derivatives of strain S25. In comparison with the parent strain S25, the cured derivative lost its ability to nodulate the host plant. Loss of the plasmid reduced significantly the strain's tolerance to acid, nitrous, and multiple antibiotics. The properties of the cured strain also indicated that the plasmid was involved in carbon and nitrogen metabolism. Reintroduction of the plasmid from S25 in the cured derivative restored its original biological phenotypes. PMID- 9735316 TI - Effect of integration host factor of RNA II synthesis in replication of plasmid containing orip 15A. AB - The synthesis rates of the replication control RNAs of plasmid orip15A. RNA I, an inhibitor of replication, and RNA II, the primer, have been determined using lacZ fusion plasmids, hybridization assay, and reverse transcription polymerase chain reaction (RT-PCR) in Escherichia coli integration host factor-positive (IHF+) and -negative (IHF-) strains containing pACYC184 plasmid (orip15A). In the absence of IHF (E. coli IHF-), expression of the lacZ gene from the PRNAII promoter increased by a factor of 4 compared with the E. coli wild type (IHF+). Also, the increase in expression was more pronounced when the IHF protein was mutated in the ihfB gene than in the ihfA gene. For the PRNAII promoter of oripMB1 (pBR322), no significant differences were found in expression of the lacZ gene in he E. coli strains examined. The level of beta-galactosidase expression from the PRNA promoter of orip 15A shows that the absence of functional IHF in the transformed strains has no effect on expression of the lacZ gene. The synthesis RNA II:RNA I ratio obtained in hybridization assays was 2.4 for E. coli IHF+ and 4.4 for E. coli IHF-. Densitometric analysis of RT-PCR products indicates that the relative levels of RNA I in E. coli IHF+ and IHF-, are equal, but the relative level of RNA II in E. coli IHF is about four times higher than in E. coli IHF+. These results indicate that the IHF protein inhibits transcription from the PRNAII promoter of orip15A plasmid. PMID- 9735318 TI - A versatile prokaryotic cloning vector with six dual restriction enzyme sites in the polylinker facilitates efficient subcloning into vectors with unique cloning sites. AB - In large and complex vectors a single restriction enzyme recognition site may be available for introduction of additional DNA requiring the development of linker fragments to create compatible insertion sites. This technology can be time consuming and costly. We describe the construction of a simple phagemid, pSFI, with a polylinker that contains six pairs of dual, rare-cutting, restriction enzyme recognition sites (NotI, SpeI, EcoRV, PstI, SacII, EagI) with multiple unique sites between each pair. This has permitted rapid subcloning of DNA with creation of single flanking restriction enzyme sites. pSFI was used to expedite transfer of viral genes to a LacZ-inducible expression vector and to an adenovirus expression cassette for production of replication-defective virus. The use of this phagemid has facilitated complex vector manipulations and is a valuable adjunct to the family of multifunctional cloning vectors. PMID- 9735319 TI - Characterization of pUCD5000 involved in pink disease color formation by Pantoea citrea. AB - Pantoea citrea, the causal agent of pink disease of pineapple, harbors a cryptic plasmid of 5229 bp. designated pUCD5000. On the basis of nucleotide and amino acid sequence analyses, pUCD5000 contains both replication and mobilization loci (bom and mobCABD) that are similar to those in plasmids pSW100 and pSW200 in Pantoea stewartii and pEC3 in Erwinia carotovora subsp. carotovora. The survival of P. citrea on pineapple does not depend on pUCD5000. However, full pink coloration development, which is characteristic of the pink disease, appears to require this plasmid. PMID- 9735320 TI - Disassembly of the post-termination complex and reduction of translational error by ribosome recycling factor (RRF)-A possible new target for antibacterial agents. AB - In this paper, we briefly review RRF (ribosome recycling factor, previously called ribosome releasing factor) (for recent reviews covering historical background see (1, 2)). PMID- 9735321 TI - Neuropeptide specificity and inhibition of recombinant isoforms of the endopeptidase 3.4.24.16 family: comparison with the related recombinant endopeptidase 3.4.24.15. AB - Endopeptidase EC 3.4.24.16 (EP24.16c, neurolysin) and thimet oligopeptidase EC 3.4.24.15 are close related members of a large family of metalloproteases. Besides their cytosolic and membrane bound form, endopeptidase EC 3.4.24.16 appears to be present in the inner membrane of the mitochondria (EP24.16m). We have overexpressed two porcine EP24.16 isoforms in E. coli and purified the recombinant proteins to homogeneity. We show here that these peptidases hydrolyse a series of neuropeptides with similar rates and at sites reminiscent of those elicited by classically purified human brain EP24.16c. All neuropeptides, except neurotensin, were similarly cleaved by recombinant endopeptidase 3.4.24.15 (EP24.15, thimet oligopeptidase), another zinc-containing metalloenzyme structurally related to EP24.16. These two EP24.16 isoforms were drastically inhibited by Pro-Ile and dithiothreitol and remained unaffected by a specific carboalkyl inhibitor (CFP-AAY-pAb) directed toward the related EP24.15. The present purification procedure of EP24.16 should allow to establish, by mutagenesis analysis, the mechanistic properties of the enzyme. PMID- 9735322 TI - Influence of the mode of insertion of SIV peptides into membranes on the structure of model membrane as studied by 31P NMR. AB - The influence on model membrane organization of a fusion peptide of SIV and of a nonfusogenic mutant of this peptide was examined by molecular modeling and by 31P NMR. The calculated mode of insertion of the fusion peptide shows that it adopts an oblique orientation towards the lipid-water interface and that this fusion peptide induces a destabilization of the bilayer structure of multilamellar vesicles as evidenced by 31P NMR observations. The SIV mutant showing a more vertical insertion into lipid layers is unable to induce nonlamellar structures. This study reinforces the correlation between fusogenic activity, induction of structures not organized in extended bilayers, and calculated mode of insertion of peptides into lipid layers. PMID- 9735323 TI - Mechanism of protein A-induced amelioration of toxicity of anti-AIDS drug, zidovudine. AB - Long-term treatment with 3-azido-3-deoxy thymidine (AZT) is often associated with myelosuppression. In AZT-treated Swiss mice, similar toxicological manifestations in terms of reduction of red blood and white blood cell counts and hemoglobin content had been observed as in AZT-treated AIDS patients. Pretreatment of animals with Protein A (PA) of Staphylococcus aureus Cowan I (1 microgram/ml), twice a week for two weeks, alleviated such hematopoietic toxicity due to AZT. AZT-induced reduction in colony-forming unit-erythroid (CFU-E) and colony-forming unit-granulocyte monocyte (CFU-GM) were also reversed by the combined treatment of AZT and PA. PA treatment showed an increased level of erythropoietin in the blood plasma, and cellularity of spleen, thymus, and bonemarrow was also increased in the group receiving combined treatment (PA+AZT), higher than that in the AZT group. AZT or its metabolites inhibited the activities of liver microsomal monooxygenases, which, however, could be regenerated in an accelerated manner by pretreatment of mice with PA. Moreover, the PA-treated group showed an accelerated clearance of AZT and/or its metabolites. These results suggest that such an immunopharmacologic approach might substantially reduce the toxic effects of drugs, such as AZT. PMID- 9735324 TI - Expanded glutamine repeat enhances complex formation of dentatorubral pallidoluysian atrophy (DRPLA) protein in human brains. AB - The genetic defect in dentatorubral-pallidoluysian atrophy (DRPLA) is expansion of the CAG repeat. The mutant gene is translated into the protein which carries the expanded glutamine repeat. Immunoblots of human brain tissues with and without reduction show that the DRPLA protein is a disulfide-bond complex and that more of this complex is formed in DRPLA brains than in control brains. This suggests that DRPLA protein undergoes greater complex formation in DRPLA brains and the expanded glutamine repeat may enhance complex formation of untruncated DRPLA protein in DRPLA brains. Immunohistochemical findings show that DRPLA protein is localized in the cytoplasm of the neuron, evidence that it undergoes rare disulfide bonding there. PMID- 9735325 TI - C-Src activation by ErbB2 leads to attachment-independent growth of human breast epithelial cells. AB - Nontumorigenic human mammary epithelial cells (184.A1 line) were stably transfected with ErbB2 or with Ha-Ras. Transformation with ErbB2, but not ras, resulted in a 5-6 fold increase in c-src activity without affecting c-src content of cells. Similar activation of c-src by ErbB2 was also observed in other non tumorigenic mammary epithelial cells, including the human line MCF10A and the mouse line NMuMG. Activation of c-src appeared to be dependent on active ErbB2 tyrosine kinase, as the ErbB2 inhibitor tyrphostin AG 825 blocked the induction of c-src kinase activity, as well as the ability of transformed cells to grow on soft agar, but not plastic. The src-selective inhibitor PP1 effectively reduced c src activity, as well as growth of ErbB2-transformed cells on soft agar, but not on plastic. These results indicate that activation of c-src is a consequence of ErbB2 kinase activity in human breast cancer cells overexpressing ErbB2, and that increased activity of c-src may be responsible for attachment-independent growth of the cells. PMID- 9735326 TI - LonR9 carrying a single Glu614 to Lys mutation inhibits the ATP-dependent protease La (Lon) by forming mixed oligomeric complexes. AB - An unusual lon mutation (called lonR9) is dominant over the wild-type gene, which encodes the ATP-dependent protease La (Lon) in Escherichia coli, when present in multicopy plasmids. Here, we cloned and sequenced lonR9, and showed that the mutant gene carries a single point mutation in its open reading frame, which leads to replacement of Glu614 by Lys. The LonR9 protein and its poly-His-tagged form were purified to apparent homogeneity. Both of the purified proteins were capable of inhibiting the ATP-dependent proteolysis and the protein-activated ATP hydrolysis by protease La. Furthermore, the His-tagged LonR9 protein was found to form mixed oligomeric complexes with protease La, upon analysis by chromatography on a metal-chelating column. These results suggest that the phenotypic dominance of the lonR9 mutant is due to the formation of mixed oligomeric complexes between LonR9 and protease La, in which the defective components prevent the function of the wild-type subunits. PMID- 9735327 TI - Reactivity of the flavin semiquinone of nitric oxide synthase in the oxygenation of arginine to NG-hydroxyarginine, the first step of nitric oxide synthesis. AB - Nitric oxide synthase (NOS) is a heme protein that catalyzes the oxygenation of L arginine in the presence of NADPH to form nitric oxide, L-citrulline and NADP+, and proceeds via two partial reactions: 1) L-Arginine --> NG-hydroxy-L-arginine 2) NG-Hydroxy-L-arginine --> L-citrulline + nitric oxide Calmodulin, FAD, FMN and tetrahydrobiopterin are required for both reactions. Reactions 1 and 2 require the input of 2 and 1 electron equivalents, respectively. Under normal multiple turnover conditions, these electrons are ultimately derived from NADPH. We previously reported that NOS contains an endogenous reductant that, in the absence of NADPH, can support the single-turnover oxygenation of L-arginine to NG hydroxy-L-arginine and a relatively small amount of L-citrulline [Campos, K. L., Giovanelli, J., and Kaufman, S. (1995) J. Biol. Chem. 270, 1721-1728]. This reductant has now been identified as the stable flavin semiquinone free radical (FSQ). Its oxidation appears to be coupled to the formation of NG-hydroxy-L arginine and L-citrulline. The rate of FSQ oxidation is two orders of magnitude slower than the flux of electrons from NADPH through NOS during normal turnover of the enzyme, indicating that FSQ is not the proximal electron donor for heme under these conditions. PMID- 9735328 TI - Increased MAPK activity and MKP-1 overexpression in human gastric adenocarcinoma. AB - Mitogen-activated protein kinase (MAPK) has been known to play a critical role in the regulation of the carcinogenesis in human cancers. In an effort to understand the functional role of the MAPK in the carcinogenesis of human gastric tissues, we examined the changes of MAPK levels in human gastric adenocarcinoma. We found that increased MAPK activity was accompanied by overexpression of mitogen activated protein kinase phosphatase-1 (MKP-1), suggesting that signaling pathways leading to the activation of MAPK and the induction of MKP-1 expression are associated with carcinogenesis of human gastric adenocarcinoma. PMID- 9735329 TI - 1,25-Dihydroxyvitamin D3 targets PKC-betaII but not PKC-alpha to the basolateral plasma membranes of rat colonocytes. AB - Prior studies by our laboratory have shown that 1, 25-dihydroxyvitamin D3 activated PKC-alpha, but not PKC-delta, -epsilon, or -zeta, in normal rat colonocytes. In the present studies we demonstrate for the first time that this secosteroid also activated PKC-betaII, another DAG- and Ca2+-dependent PKC isoform recently shown to be present in these cells. Moreover, this activation of PKC-betaII by 1,25-dihydroxyvitamin D3 treatment of isolated colonocytes was shown to be lost in cells from vitamin D-deficient rats and, at least partially, restored by repleting these animals with this secosteroid for 7 days. Under basal conditions, the expression of PKC-alpha and -betaII in brush-border membranes was comparable to their respective expression in basolateral plasma membranes of rat colonocytes. In contrast, the expression of PKC-delta was significantly greater in brush-border membranes, whereas PKC-epsilon and -zeta were enriched in the basolateral plasma membranes. Furthermore, 1,25-dihydroxyvitamin D3 specifically induced the translocation of PKC-betaII, but not PKC-alpha, to the basolateral, but not brush-border plasma membranes of rat colonocytes, via a pp60(c-src) dependent mechanism. PMID- 9735330 TI - Resveratrol arrests the cell division cycle at S/G2 phase transition. AB - Resveratrol (3,5,4'-trihydroxystilbene) is a naturally occurring phytoalexin, found in grapes and wine, which has been reported to exert a variety of important pharmacological effects. We have investigated the activity of resveratrol on proliferation and differentiation of the promyelocitic cell line HL-60. A concentration as low as 30 microM causes a complete arrest of proliferation and a rapid induction of differentiation towards a myelo-monocytic phenotype. Analyses by flow cytometry showed the absence of the G2/M peak and the accumulation of cells in G1 and S phases. Moreover, at the concentrations employed, a very low amount of apoptotic cells was evidenced. A detailed biochemical analysis demonstrated that the G1 phase of the cell division cycle engine was completely unmodified by resveratrol addition, thus indicating that the G1 --> S transition occurs normally. Conversely, after only 24 h treatment, a significant increase of cyclins A and E could be observed along with the accumulation of cdc2 in the inactive phosphorylated form. These data demonstrate that resveratrol causes a complete and reversible cell cycle arrest at the S phase checkpoint. PMID- 9735331 TI - Residual bodies stimulate rat Sertoli cell plasminogen activator activity. AB - Previous studies suggested indirectly that residual bodies (RB) may play a role in the regulation of the proteolytic system within the testis and in the coordination of the spermatogenetic process. In the present study, we examined the effects of RB recovered from adult rat testes by centrifugal elutriation on Sertoli cell plasminogen activator (PA) levels, by zymography and ELISA procedures. Addition of RB to Sertoli cell cultures prepared from 20-day-old rat testes resulted in a dramatic stimulation of PA. Effects were dose- and time dependent. Phagocytosis of RB by Sertoli cells leads to a rapid stimulation of Sertoli cell interleukin-1alpha (IL-1alpha), a cytokine potentially involved in the regulation of spermatogenesis; the effects of IL-1alpha were investigated. We found that IL-1alpha augmented PA levels and that immunodepletion of Sertoli cell RB cocultures with anti-IL-1alpha antibodies abrogated the stimulatory effects of RB on PA. Together, the present findings indicate that RB enhance Sertoli cell PA and that IL-1alpha may be involved in that control. PMID- 9735332 TI - JC9813-A putative novel human papillomavirus identified by PCR-DS. AB - Papillomaviruses consist of more than 130 viral types described so far. Most of them are human papillomaviruses (HPV) of supergroup A, demonstrating ano-genital tropism and characterized as etiological agents for benign and malignant cervical lesions in women. A PCR-direct sequencing (PCR-DS) approach with P-33 labeled dideoxynucleotides was used to detect and type human papillomaviruses in cervical biopsies. One novel sequence was identified in a LSIL (low-grade squamous intraepithelial lesions) specimen from an HIV-positive English Canadian patient. The structure of the viral gene L1 was determined, yielding a putative novel HPV type of supergroup A (clade A8) named JC9813. PMID- 9735333 TI - Cloning and characterization of a new subtype of thyrotropin-releasing hormone receptors. AB - A new subfamily member of thyrotropin releasing hormone (TRH) receptor gene, TRHR2, was isolated from rat brain cDNAs. The deduced amino acid sequence of TRHR2 is 51 % identical to that of rat TRH receptor gene which was reported previously. Northern blot analysis with TRHR2 probe revealed brain-specific expression of a 9.5 kb mRNA. In a binding experiment using the TRHR2-expressing COS cells, specific binding of TRH to TRHR2 was observed with Kd value of 9 nM which was equivalent to the Kd value (= 13 nM) of TRH binding to the TRH receptor previously reported. The active metabolite of TRH, histidyl-proline diketopiperazine, or cyclo(His-Pro), showed no specific binding activity. These results suggest that TRHR2 is a novel subtype of TRH receptor. PMID- 9735334 TI - Genetic deletion of AT2 receptor antagonizes angiotensin II-induced apoptosis in fibroblasts of the mouse embryo. AB - To examine whether angiotensin II (Ang II) can trigger apoptosis via Ang II type 2 (AT2) receptor, two genotypes of skin fibroblasts cultured from the AT2 receptor gene targeting homozygous (AT2-/-) and wild-type (AT2+/+) mouse embryos, respectively, were studied when exposed to Ang II. In the AT2+/+ fibroblasts, mRNA expression of the AT2 receptor was modulated by Ang II in a dose-dependent manner and apoptosis appeared with the convincing features of internucleosomal DNA fragmentation and DNA content decrease after stimulation with Ang II, whereas Ang II had no significant impact on the AT2-/- fibroblasts due to the AT2 receptor gene deletion. This is the first report using a gene targeting study to demonstrate that Ang II induces apoptosis through the AT2 receptor in the fibroblasts of the mouse embryo. PMID- 9735335 TI - Insulin down-regulates angiotensinogen gene expression and angiotensinogen secretion in cultured adipose cells. AB - Adipose tissue is an important source of angiotensinogen (AT) after liver. Since an association exists between body mass index, hypertension, and insulin resistance, the role of insulin on the regulation of AT gene expression and AT secretion was examined in cultured Ob1771 and 3T3-F442A adipose cells. Within a physiological range of concentrations (1-17 nM), insulin exerted a negative effect on the abundance of AT mRNA and the secretion of AT. Alterations of insulin-resistance by treatment of adipose cells with TNF-alpha or the thiazolidinedione BRL49653 led respectively to a decrease or an increase in the potency of insulin to down-regulate AT gene expression, whereas maximal inhibition by insulin increased from 30% in TNFalpha-treated cells to 60% in BRL49653-treated cells. These results suggest that a potential link between insulin resistance and high blood pressure may exist by means of increased AT secretion from adipose tissue, especially in obese subjects. PMID- 9735336 TI - Circadian oscillation of BMAL1, a partner of a mammalian clock gene Clock, in rat suprachiasmatic nucleus. AB - A superfamily gene which encodes a bHLH (basic helix-loop-helix)/PAS transcription factor, BMAL1, was cloned and sequenced from rat cDNA. A robust circadian rhythm of rat BMAL1 expression was detected by in situ hybridization in the suprachiasmatic nucleus (SCN), the site of the circadian clock, with the highest level at the subjective night. Less prominent and completely reversed circadian rhythms of rBMAL1 mRNA were observed in the piriform and parietal cortices. The hybridization signals of rBMAL1 mRNA were also detected in the olfactory bulb, hippocampus, and cerebellum. Since the product of rBMAL1 was recently demonstrated to dimerize with the protein of a mammalian clock gene, Clock, and the protein complex was shown to bind the E Box in the promoter region of mPer1 (a mouse homologue to Drosophila clock gene, Per), rBMAL1 possibly plays a critical role in the clock mechanism generating the circadian oscillation in rats. PMID- 9735337 TI - Phytosphingosine biosynthesis differs from sphingosine in fish leukocytes and involves a transfer of methyl groups from [3H-methyl]methionine precursor. AB - We have studied the incorporation of radioactivity from either [3-3H]serine as the direct or [3H-methyl]methionine as the indirect precursor into sphingoid bases of free ceramides in lymphocytes from fish. Radioactivity from serine was incorporated mostly in the sphingosine moiety of ceramides. In contrast, the radioactivity from methionine was exclusively incorporated into phytosphingosine base (i.e., 4-hydroxy-sphinganine) and the incorporation increased by about twofold in the presence of folic acid or niacinamide. Identity of the long-chain bases, phytosphingosine and sphingosine, was established chemically by thin-layer chromatography, chemical degradation, and gas-liquid chromatography. PMID- 9735338 TI - Renin inhibits the vasorelaxation induced by nitroso albumin. AB - Nitrosated proteins exhibit actions characteristic of free NO. As the vasorelaxation effect of nitrosated albumin is rapidly inactivated in plasma, we postulated that a protease could remove or modify the NO attached to albumin. We found that the ability of plasma to inactivate the vasorelaxing action of NO bovine serum albumin (NO-BSA) is restricted to a plasma fraction containing macromolecules. We also found that a crude preparation of renal renin also inactivated the vasorelaxation action of NO-BSA and UV-spectrophotometric analysis showed that the 335-nm signal of NO-BSA was significantly decreased by renin. This decrease could be prevented by a renin inhibitor or by immunodepleting the renin preparation with a monoclonal antibody to renin. The data suggest that renin accelerates the uncoupling of NO to albumin. Such a function may be important in the control of vascular tone and blood pressure. PMID- 9735339 TI - Lipolytic effect of in vivo leptin administration on adipocytes of lean and ob/ob mice, but not db/db mice. AB - The present study has examined the effect of a single in vivo intraperitoneal injection of the adipocyte-derived hormone, leptin, on the in vitro lipolysis of fat cells of different types of mice. Administration of 1 and 10 mg leptin per kg body weight to ob/ob mice significantly increased (P < 0.0001) the basal lipolytic activity compared to ob/ob mice receiving vehicle solution (phosphate buffered saline, PBS). The highest leptin dose tested (10 mg/kg body weight) produced a threefold increase in basal lipolysis. In lean mice administration of 10 mg leptin per kilogram of body weight produced an increase in basal lipolysis of 52.7% (P < 0.01). However, in db/db mice none of the three leptin doses injected had a significant effect on the lipolytic activity of adipocytes relative to basal lipolysis observed in db/db mice injected with PBS only. These data provide evidence for a lipolytic effect of leptin on white adipose tissue, which operates independently from changes in food intake, body weight, and the size of the fat stores. PMID- 9735340 TI - Molecular identification of a role for tyrosine 167 in the function of the human intestinal proton- coupled dipeptide transporter (hPepT1). AB - hPepT1 is a proton-coupled peptide transporter that mediates the absorption of di and tripeptides. Here we show that tyrosine 167 (Y167) in transmembrane domain 5 (TMD5) of this 12-transmembrane spanning protein contributes to its transport function. We identified this particular amino acid by a computer model of the arrangement of the TMDs of hPepT1 and investigated its role by site-directed mutagenesis and dipeptide uptake studies. [3H]Gly-sar uptake in cells transiently transfected with Y167A-hPepT1 was abolished completely, even though the level of Y167A-hPepT1 expression by Western blot analysis and cell surface expression by immunofluorescence microscopy was similar to those of the wild type. Therefore, mutation affected transport function, but apparently not the steady-state protein level or trafficking of the transporter to the plasma membrane. Moreover, mutation of Y167 into phenylalanine, serine, or histidine all abolished gly-sar uptake in transfected HEK 293 cells. Taken together, these findings suggest that Y167 plays an essential role in hPepT1 function, perhaps due to the unique chemistry of its phenolic side chain. PMID- 9735341 TI - Nitric oxide response to shear stress by human bone cell cultures is endothelial nitric oxide synthase dependent. AB - Bone cells, in particular osteocytes, are extremely sensitive to shear stress, a phenomenon that may be related to mechanical adaptation of bone. In this study we examined whether human primary bone cells produce NO in response to fluid shear stress and established by RT/PCR which NOS isoforms were expressed before and after application of shear stress. One hour pulsating fluid flow (PFF; 0.7 +/- 0.02 Pa, 5 Hz) caused a rapid (within 5 min) 2 to 4-fold increase in NO production. NO release was only transiently increased during the first 15 min of exposure to PFF, and remained at control levels during a 1-24 hr postincubation period. In both control and PFF-treated cells, mRNA was easily detected for ecNOS, but not nNOS, and only minimal amounts iNOS were found. mRNA levels for ecNOS increased 2-fold at 1 hr after 1 hr PFF treatment. These results suggest that the rapid production of NO by human bone cells in response to fluid flow results from activation of ecNOS. PFF also leads to an increase in ecNOS mRNA which is likely related to the shear stress responsive element in the promoter of ecNOS. PMID- 9735342 TI - Hsc62, a new DnaK homologue of Escherichia coli. AB - We have cloned and expressed the ORF o170#1 of Escherichia coli, which encodes a 62-kDa protein sharing 33% homology in primary structure with DnaK and Hsc66, Hsp70 homologues of E. coli. The purified gene product, which we named Hsc62, clearly showed ATPase activity and was bound to a gelatin-agarose gel, from which it was specifically eluted with ATP magnesium salt. Thus, Hsc62 is similar to DnaK in this respect and probably functions as a molecular chaperon in E. coli. However, Hsc62 differs markedly from DnaK and also from Hsc66 in response to temperature: the optimum temperature for ATPase activity was increased stepwise in the order of Hsc62, Hsc66, and DnaK. Hsc66 is activated by DnaJ of E. coli in the same manner as DnaK, the natural partner protein of DnaJ. However, Hsc62 is distinct from the others: the ATPase activity of Hsc62 was not elevated by DnaJ. PMID- 9735343 TI - Tenascin suppresses CD3-mediated T cell activation. AB - Tenascin (TN) is an extracellular matrix protein which interferes with fibronectin (FN)-dependent cell attachment and activation as a natural antagonist to FN action. In this study, we examined the inhibitory effect of TN on T cell proliferation induced by immobilized anti-CD3 Ab combined with various costimulators in a serum-free condition. Consistent with previous studies, human T cell activation induced by anti-CD3 plus FN was completely inhibited by the addition of TN. Interestingly, TN could interfere with T cell proliferations costimulated by various very late activation antigen (VLA) integrin/ligand interactions such as VLA-4/FN, VLA-5/FN and VLA-6/laminin. Furthermore, lymphocyte function-associated antigen 1 (LFA-1)-, CD2- and CD28-costimulated T cell activation were also inhibited by TN, while TN could not affect the phorbol ester-stimulated T cell proliferation. Collectively, TN inhibits anti-CD3-induced T cell proliferation irrespectively of costimulatory molecules, suggesting that TN acts as a generally immunosuppressive extracellular matrix protein which potentially interferes with T cell receptor/CD3-mediated T cell activation. PMID- 9735344 TI - Evaluation of Lama5 as a candidate for the mouse ragged (Ra) mutation. AB - The laminin alpha5 chain is a component of the basement membranes of many developing and adult tissues. The mouse laminin alpha5 chain gene (Lama5) has been mapped close to the locus of the semidominant ragged (Ra) mutation on distal chromosome 2. The cause of the Ra mutation, which is usually lethal in the homozygous state, has not been determined. We have investigated whether a defect in Lama5 is responsible for the ragged mutation, using the RaJ strain. No differences in the level of the laminin alpha5 chain transcript were found in placental RNA from homozygous RaJ mutant embryos compared to normal littermates. Antiserum raised against a recombinant laminin alpha5 chain polypeptide stained the basement membranes of both normal and homozygous mutant embryos to a similar extent. More precise mapping of Lama5 on an interspecific Ra backcross indicated that Lama5 is proximal to the Ra locus. These results exclude Lama5 as a candidate gene for the Ra mutation. PMID- 9735345 TI - Surfactant protein A (SP-A) forms a novel supraquaternary structure in the form of fibers. AB - We have found by transmission electron microscopy that bovine surfactant protein A (SP-A) formed extended fibers in the presence of calcium. On phosphatidylcholine or especially dipalmitoylphosphatidylcholine monolayers, SP-A at roughly 0.005 mg/ml formed large numbers of fibers and elaborate fibrous networks. This observation suggested that the weak protein:protein interactions amongst free SP-As could be stabilized by phospholipids. In the presence of glycolipid GM1-ganglioside, SP-A's globular headgroup regions appeared enlarged and only small non-fibrous clusters were observed. PMID- 9735346 TI - Functional effects of FGF-13 on human lung fibroblasts, dermal microvascular endothelial cells, and aortic smooth muscle cells. AB - We studied the effects of FGF-13 and FGF-2 on human lung fibroblasts, dermal microvascular endothelial cells, and aortic smooth muscle cells. FGF-13 induced cell growth of lung fibroblasts and aortic smooth muscle cells but had no effect on dermal vascular endothelial cells. FGF-2 induced cell growth in all the three cell types. FGF-13 and FGF-2 had little effect on IL-6 production by lung fibroblasts and aortic smooth muscle cells and substantially enhanced that induced by IL-1alpha. In contrast, FGF-13 and FGF-2 had little effect on IL-6 production by dermal vascular endothelial cells, either alone or in synergy with IL-1alpha. PMID- 9735347 TI - Structural and functional analysis of the putative inositol 1,3,4, 5 tetrakisphosphate receptors GAP1(IP4BP) and GAP1(m). AB - Previously we have purified and cloned a high affinity isomerically specific inositol 1,3,4,5-tetrakisphosphate (Ins(1,3,4,5)P4)-binding protein which, because it is clearly a member of the GAP1 family of Ras GTPase-activating proteins (GAP), we have termed GAP1(IP4BP). Here we show that expressed full length GAP1(IP4BP) binds Ins(1,3,4, 5)P4 with an affinity and specificity similar to that of the originally purified protein, a binding activity which is dependent on a functional PH/Btk domain. Furthermore, we highlight a fundamental distinction between GAP1(IP4BP) and its homologue GAP1(m), namely that both proteins function as Ras GAPs but only GAP1(IP4BP) displays Rap GAP activity. PMID- 9735348 TI - In vivo detection of mutations induced by aflatoxin B1 using human CYP3A7/HITEC hybrid mice. AB - CYP3A7-M10 mouse is a transgenic mouse carrying human CYP3A7 cDNA, in which CYP3A7 is expressed in the small intestine but not in the kidney. HITEC mouse is a transgenic mouse developed to detect mutagenic potency of various chemicals in vivo. The M10/HITEC mouse was established by crossmating of these two strains of mice. When a 9,000 x g supernatant fraction prepared from the small intestine was added to an incubation mixture for Ames test with Salmonella typhimurium TA98 strain to examine the mutagen-producing activity from aflatoxin B1 (AFB1), the mutagen-producing activities of the 9, 000 x g supernatant fraction from the small intestine was found to be 1.7-fold higher in the M10/HITEC mice than in HITEC mice. Such a difference in the capacity to activate AFB1 was not seen with the 9, 000 x g supernatant fraction from the kidney from both strains of mice. Male M10/HITEC mice of 8 weeks old were treated with a single i.p. injection of AFB1 ( 8 mg/kg body weight). The mutation of the introduced rpsL gene in the genomic DNA from the small intestine and the kidney was analyzed. The mutation frequency in the small intestine of M10/HITEC mice was significantly higher (p<0.05) than that of HITEC mice, while the mutation frequency in both strains was similar in the kidney. These results provide the first evidence for the toxicological function of CYP3A7 in vivo. PMID- 9735349 TI - Characterization of phospholipase A2 (PLA2) from Taiwan Cobra: isoenzymes and their site-directed mutants. AB - Extracellular and secretory phospholipase A2 (PLA2), a class of phospholipid digesting enzyme, is widely distributed in animal venoms of reptiles and insects. Two cDNAs encoding PLA2 isoenzymes from Taiwan Cobra (Naja naja atra) were cloned into pQE-30 plasmid vector and expressed in Escherichia coli. The recombinant products were subjected to refolding using sulfonation under reduction/oxidation conditions with glutathione and enterokinase removal of His-tag, resulting in the active recombinant PLA2 with the same molecular masses of native enzymes as determined by mass spectrometry. The recombinant PLA2 was also shown by circular dichroism to possess a secondary structure similar to native PLA2. The enzymatic activity of the major isoenzyme (PLA2-1) is higher than the other minor isoenzyme (PLA2-2), which shows two amino acid difference from PLA2-1. Site-directed mutagenesis was used to probe the structure/function relationship of two highly conserved residues among all reported PLA2, i.e., His-47 and Asp-93. Replacement of His-47 residue by either Ala or Arg resulted in the complete loss of activity. Similarly, the mutant Asp-93 --> Asn (D93N) also retained little activity. These results suggest that both His-47 and Asp-93 are essential for the catalytic activity of PLA2. Computer graphic study, based on homology modelling, highlights the differences between native PLA2 isoenzymes and their site-directed mutants, which may account for the differences in the observed biological activity. PMID- 9735350 TI - An Arabidopsis gene family encoding DRE/CRT binding proteins involved in low temperature-responsive gene expression. AB - In higher plants, a cis-acting element, DRE/CRT, is involved in gene expression responsive to drought and low-temperature stress. To understand signal transduction pathways from the cold stress signal to gene expression, we characterized a gene family for DRE/CRT-binding proteins DREB1A and CBF1 in Arabidopsis thaliana. DREB1A and CBF1 were shown to be involved in low temperature-responsive gene expression. We screened an Arabidopsis genomic DNA library with the cDNA fragment of DREB1A as a probe and isolated DREB1A and 2 related genes, DREB1B (= CBF1) and DREB1C. These were arrayed in the order B, A, C in an 8.7 kb region of Arabidopsis chromosome 4. Northern blot analysis using gene-specific probes showed that the 3 DREB1 genes are induced mainly by cold stress but not by osmotic stress in leaves, roots, and stems. Several conserved sequences were found in the promoter regions of all 3 genes. The beta glucuronidase (GUS) reporter gene driven by the DREB1 promoters was induced at transcriptional level by low temperature in transgenic Arabidopsis plants. PMID- 9735351 TI - High cleavage activity and stability of hammerhead ribozymes with a uniform 2' amino pyrimidine modification. AB - The uniform 2'-pyrimidine modifications have been found to inhibit ribozyme cleavage activity. However, in the present study we show that a good cleavage activity can be achieved for the 2'-amino modified ribozymes when their sequences were designed to contain only a few pyrimidines in helix I. In particular, ribozymes with no pyrimidines in helix I cleaved their target RNAs with almost the same efficacy as their unmodified versions. Interestingly, selective 2'-amino modification at positions 2.1 and 2.2 reduced the ribozyme cleavage activity by 8 fold, suggesting that the 2'-amino groups at these two positions may interfere with the formation of the ribozyme active conformation. In addition, uniformly modified ribozymes showed a remarkable stability in serum. Taken together these results should facilitate the design of stable ribozymes with sustained cleavage activity. PMID- 9735352 TI - Erx, a novel retina-specific homeodomain transcription factor, can interact with Ret 1/PCEI sites. AB - Our previous studies on the transcriptional regulation of rod opsin gene expression had defined a strikingly conserved element, Ret 1/PCEI, present in the upstream regulatory regions of opsin and other photoreceptor-specific genes. This element interacts with a 40 kDa, developmentally regulated, retina-specific protein. In this study we report the cloning of the novel retina-specific homeodomain protein Erx. Erx contains a homeodomain that is 79% homologous to that of Drosophila empty spiracles. This 40 kDa protein can interact with the Ret 1 element in electrophoretic mobility shift assays. Mutation of key residues in Ret 1 eliminates all Erx binding. Transient transfection of Y79 retinobalstoma cells with Erx leads to significant transcriptional activation of a reporter gene via Ret 1 elements. We conclude that Erx is the Ret 1 binding activity. This is the first example of a Q50 homeodomain protein expressed in retinal photoreceptors. PMID- 9735353 TI - Protein kinase B is expressed in pancreatic beta cells and activated upon stimulation with insulin-like growth factor I. AB - Protein kinase B (PKB) is involved in signaling to a multitude of important cellular events and is activated by insulin and growth factors, including insulin like growth factor I (IGF-I). We show here expression of PKB in pancreatic islets and in the beta cell lines HIT-T15, INS-1, and RINm5F. Expression of PKB mRNA and the presence of PKB isoforms (alpha, beta, and gamma) were assessed by Northern blot analysis and RT-PCR, respectively. Antibodies recognizing different parts of PKB isoforms were employed to demonstrate PKB protein expression by immunoblot analysis. By use of immunohistochemistry in rat and mouse pancreatic tissue sections, PKB was localized to predominantly beta cells. Regulation of PKB was examined in INS-1 and RINm5F cells; upon stimulation with IGF-I (5-10 min), PKB was phosphorylated and activated (approximately 3-fold) by a wortmannin-sensitive mechanism, indicating involvement of phosphatidylinositol-3 kinase. The possible participation of PKB in signal transduction pathways modulating cAMP-dependent insulin secretion and in proliferation of beta cells is discussed. PMID- 9735354 TI - Lhx2, a vertebrate homologue of apterous, regulates vertebrate limb outgrowth. AB - apterous specifies dorsal cell fate and directs outgrowth of the wing during Drosophila wing development. Here we show that, in vertebrates, these functions appear to be performed by two separate proteins. Lmx-1 is necessary and sufficient to specify dorsal identity and Lhx2 regulates limb outgrowth. Our results suggest that Lhx2 is closer to apterous than Lmx-1, yet, in vertebrates, Lhx2 does not specify dorsal cell fate. This implies that in vertebrates, unlike Drosophila, limb outgrowth can be dissociated from the establishment of the dorsoventral axis. PMID- 9735355 TI - Requirement for Brn-3c in maturation and survival, but not in fate determination of inner ear hair cells. AB - Mutations in the POU domain gene Brn-3c causes hearing impairment in both the human and mouse as a result of inner ear hair cell loss. We show here that during murine embryogenesis, Brn-3c is expressed in postmitotic cells committed to hair cell phenotype but not in mitotic progenitors in the inner ear sensory epithelium. In developing auditory and vestibular sensory epithelia of Brn-3c-/- mice, hair cells are found to be generated and undergo initial differentiation as indicated by their morphology, laminar position and expression of hair cell markers, including myosins VI and VIIa, calretinin and parvalbumin. However, a small number of hair cells are anomalously retained in the supporting cell layer in the vestibular sensory epithelia. Furthermore, the initially differentiated hair cells fail to form stereociliary bundles and degenerate by apoptosis in the Brn-3c-/- mice. These data indicate a crucial role for Brn-3c in maturation, survival and migration of hair cells, but not in proliferation or commitment of hair cell progenitors. PMID- 9735356 TI - Non-autonomous regulation of a graded, PKA-mediated transcriptional activation signal for cell patterning. AB - The pseudoplasmodium or migrating slug of Dictyostelium is composed of non terminally differentiated cells, organized along an anteroposterior axis. Cells in the anterior region of the slug define the prestalk compartment, whereas most of the posterior zone consists of prespore cells. We now present evidence that the cAMP-dependent protein kinase (PKA) and the RING domain/leucine zipper protein rZIP interact genetically to mediate a transcriptional activation gradient that regulates the differentiation of prespore cells within the posterior compartment of the slug. PKA is absolutely required for prespore differentiation. In contrast, rZIP negatively regulates prespore patterning; rzpA cells, which lack rZIP, have reduced prestalk differentiation and a corresponding increase in prespore-specific gene expression. Using cell-specific markers and chimaeras of wild-type and rzpA- cells, we show that rZIP functions non-autonomously to establish a graded, prespore gene activation signal but autonomously to localize prespore expression. Overexpression of either the catalytic subunit or a dominant-negative regulatory subunit of PKA further demonstrates that PKA lies within the intracellular pathway that mediates the extracellular signal and regulates prespore patterning. Finally, we show that a 5'-distal segment within a prespore promoter that is responsive to a graded signal is also sensitive to PKA and rZIP, indicating that it acts directly at the level of prespore-specific gene transcription for regulation. PMID- 9735357 TI - The Drosophila trithorax group proteins BRM, ASH1 and ASH2 are subunits of distinct protein complexes. AB - The trithorax group gene brahma (brm) encodes an activator of Drosophila homeotic genes that functions as the ATPase subunit of a large protein complex. To determine if BRM physically interacts with other trithorax group proteins, we purified the BRM complex from Drosophila embryos and analyzed its subunit composition. The BRM complex contains at least seven major polypeptides. Surprisingly, the majority of the subunits of the BRM complex are not encoded by trithorax group genes. Furthermore, a screen for enhancers of a dominant-negative brm mutation identified only one trithorax group gene, moira (mor), that appears to be essential for brm function in vivo. Four of the subunits of the BRM complex are related to subunits of the yeast chromatin remodeling complexes SWI/SNF and RSC. The BRM complex is even more highly related to the human BRG1 and hBRM complexes, but lacks the subunit heterogeneity characteristic of these complexes. We present biochemical evidence for the existence of two additional complexes containing trithorax group proteins: a 2 MDa ASH1 complex and a 500 kDa ASH2 complex. These findings suggest that BRM plays a role in chromatin remodeling that is distinct from the function of most other trithorax group proteins. PMID- 9735358 TI - A role for the fibroblast growth factor receptor in cell fate decisions in the developing vertebrate retina. AB - The mature vertebrate retina contains seven major cell types that develop from an apparently homogenous population of precursor cells. Clonal analyses have suggested that environmental influences play a major role in specifying retinal cell identity. Fibroblast growth factor-2 is present in the developing retina and regulates the survival, proliferation and differentiation of developing retinal cells in culture. Here we have tested whether fibroblast growth factor receptor signaling biases retinal cell fate decisions in vivo. Fibroblast growth factor receptors were inhibited in retinal precursors in Xenopus embryos by expressing a dominant negative form of the receptor, XFD. Dorsal animal blastomeres that give rise to the retina were injected with cDNA expression constructs for XFD and a control non-functional mutant receptor, D48, and the cell fates of transgene expressing cells in the mature retina determined. Fibroblast growth factor receptor blockade results in almost a 50% loss of photoreceptors and amacrine cells, and a concurrent 3.5-fold increase in Muller glia, suggesting a shift towards a Muller cell fate in the absence of a fibroblast growth factor receptor signal. Inhibition of non-fibroblast-growth-factor-mediated receptor signaling with a third mutant receptor, HAVO, alters cell fate in an opposite manner. These results suggest that it is the balance of fibroblast growth factor and non fibroblast growth factor ligand signals that influences retinal cell genesis. PMID- 9735360 TI - The Drosophila SOX-domain protein Dichaete is required for the development of the central nervous system midline. AB - SOX-domain proteins are a class of developmentally important transcriptional regulators related to the mammalian testis determining factor SRY. In common with other SOX-domain genes, the Drosophila Dichaete gene has a dynamic expression profile in the developing central nervous system, including cells of the ventral midline. We find defects in the differentiation of midline glia and concomitant axonal defects in Dichaete mutants that are rescued by driving Dichaete expression in the midline. Since Dichaete is required for the correct specification or differentiation of midline glia, we have used the ventral midline as a model system to study SOX gene function in vivo and demonstrate a genetic interaction between Dichaete and the POU domain gene ventral veinless. In mammals, a protein related to Dichaete, SOX2, also interacts with POU transcription factors. The midline phenotypes of Dichaete mutations are rescued by expression of mouse SOX2. Our data suggest that SOX gene structure, function and interactions have been conserved during evolution. PMID- 9735359 TI - Synergistic signaling by two BMP ligands through the SAX and TKV receptors controls wing growth and patterning in Drosophila. AB - In Drosophila wing discs, a morphogen gradient of DPP has been proposed to determine the transcriptional response thresholds of the downstream genes sal and omb. We present evidence that the concentration of the type I receptor TKV must be low to allow long-range DPP diffusion. Low TKV receptor concentrations result, however, in low signaling activity. To enhance signaling at low DPP concentrations, we find that a second ligand, GBB, augments DPP/TKV activity. GBB signals primarily through the type I receptor SAX, which synergistically enhances TKV signaling and is required for proper OMB expression. We show that OMB expression in wing discs requires synergistic signaling by multiple ligands and receptors to overcome the limitations imposed on DPP morphogen function by receptor concentration levels. PMID- 9735361 TI - Bix1, a direct target of Xenopus T-box genes, causes formation of ventral mesoderm and endoderm. AB - Brachyury, a member of the T-box gene family, is required for posterior mesoderm and notochord differentiation in vertebrate development, and mis-expression of Xenopus Brachyury causes ectopic mesoderm formation. Brachyury is a transcription activator, and its ability to activate transcription is essential for its biological function, but Brachyury target genes have proved difficult to identify. Here we employ a hormone-inducible Brachyury construct and subtractive hybridization to search for such targets. Using this approach we have isolated Bix1, a homeobox gene expressed both in the marginal zone of Xenopus and in the vegetal hemisphere. Expression of Bix1 is induced in an immediate-early fashion by mesoderm-inducing factors such as activin as well as by the products of the T box genes Xbra and VegT (also known as Antipodean, Brat and Xombi). Activation of Bix1 in response to Xbra is direct in the sense that it does not require protein synthesis, and both Xbra and VegT activate expression of a reporter gene driven by the Bix 5' regulatory region, which contains an Xbra/VegT binding site. Mis expression of low levels of Bix1 causes formation of ventral mesoderm, while high levels induce endodermal differentiation. These results suggest that Bix1 acts downstream of both VegT and Xbra to induce formation of mesoderm and endoderm. PMID- 9735362 TI - Atm deficiency results in severe meiotic disruption as early as leptonema of prophase I. AB - Infertility is a common feature of the human disorder ataxia-telangiectasia and Atm-deficient mice are completely infertile. To gain further insight into the role of ATM in meiosis, we examined meiotic cells in Atm-deficient mice during development. Spermatocyte degeneration begins between postnatal days 8 and 16.5, soon after entry into prophase I of meiosis, while oocytes degenerate late in embryogenesis prior to dictyate arrest. Using electron microscopy and immunolocalization of meiotic proteins in mutant adult spermatocytes, we found that male and female gametogenesis is severely disrupted in Atm-deficient mice as early as leptonema of prophase I, resulting in apoptotic degeneration. A small number of mutant cells progress into later stages of meiosis, but no cells proceed beyond prophase I. ATR, a protein related to ATM, DMC1, a RAD51 family member, and RAD51 are mislocalized to chromatin and have reduced localization to developing synaptonemal complexes in spermatocytes from Atm-deficient mice, suggesting dysregulation of the orderly progression of meiotic events. ATM protein is normally present at high levels primarily in ova cytoplasm of developing ovarian follicles, and in the nucleus of spermatogonia and to a lesser extent in spermatoctyes, but without localization to the synaptonemal complex. We propose a model in which ATM acts to monitor meiosis by participation in the regulation or surveillance of meiotic progression, similar to its role as a monitor of mitotic cell cycle progression. PMID- 9735363 TI - Muscle and tendon morphogenesis in the avian hind limb. AB - The proper development of the musculoskeletal system in the tetrapod limb requires the coordinated development of muscle, tendon and cartilage. This paper examines the morphogenesis of muscle and tendon in the developing avian hind limb. Based on a developmental series of embryos labeled with myosin and tenascin antibodies in whole mount, an integrative description of the temporal sequence and spatial pattern of muscle and tendon morphogenesis and their relationship to cartilage throughout the chick hind limb is presented for the first time. Anatomically distinct muscles arise by the progressive segregation of muscle: differentiated myotubes first appear as a pair of dorsal and ventral muscle masses; these masses subdivide into dorsal and ventral thigh, shank and foot muscle masses; and finally these six masses segregate into individual muscles. From their initial appearance, most myotubes are precisely oriented and their pattern presages the pattern of future, individual muscles. Anatomically distinct tendons emerge from three tendon primordia associated with the major joints of the limb. Contrary to previous reports, comparison of muscle and tendon reveals that much of their morphogenesis is temporally and spatially closely associated. To test whether reciprocal muscle-tendon interactions are necessary for correct muscle-tendon patterning or whether morphogenesis of each of these tissues is autonomous, two sets of experiments were conducted: (1) tendon development was examined in muscleless limbs produced by coelomic grafting of early limb buds and (2) muscle development was analyzed in limbs where tendon had been surgically altered. These experiments demonstrate that in the avian hind limb the initial morphogenetic events, formation of tendon primordia and initial differentiation of myogenic precursors, occur autonomously with respect to one another. However, later morphogenetic events, such as subdivision of muscle masses and segregation of tendon primordia into individual tendons, do require to various degrees reciprocal interactions between muscle and tendon. The dependence of these later morphogenetic events on tissue interactions differs between different proximodistal regions of the limb. PMID- 9735364 TI - Specification of first quartet micromeres in Ilyanassa involves inherited factors and position with respect to the inducing D macromere. AB - In the embryos of the gastropod Ilyanassa obsoleta, the development of several ectodermal structures requires an inductive interaction between the micromeres and the D macromere. The first quartet micromeres (1a, 1b, 1c and 1d) contribute to the head of the larva and descendants of 1a and 1c normally develop the eyes. The eyes do not develop if 1a and 1c are removed at the eight-cell stage. However, regulative eye development may occur if the precursors of 1a and 1c are removed at the two- or four-cell stage. One purpose of this study was to demonstrate which cells of the cleavage-stage embryo have the potential to develop an eye. The results of blastomere deletion experiments suggest that only the first quartet micromeres have this ability. In addition, the 1b micromere was found to be equivalent to 1a and 1c, but 1d was found to have a poorer eye forming ability. A second purpose of this study was to examine how eye development is normally restricted to the 1a and 1c micromeres. Cell transplantation experiments demonstrate that the proximity of a first quartet micromere relative to the inducing D macromere is important for determining whether or not it will go on to develop an eye. The 1b micromere may not develop an eye during normal development because it is too far from the D macromere. However, the eye-forming ability of the 1d micromere is not influenced by its close position to the D macromere, but is restricted by its polar lobe lineage. PMID- 9735365 TI - Role of Nudel protease activation in triggering dorsoventral polarization of the Drosophila embryo. AB - The establishment of embryonic dorsoventral polarity in Drosophila depends on a signaling mechanism in which the signal for ventral development is locally produced. This mechanism requires the activity of the nudel gene in ovarian follicle cells, which provide dorsoventral positional information for the embryo. The nudel gene product, a large mosaic protein with a central serine protease domain, has been proposed to function in locally triggering a protease cascade that produces the ventral signal. Here we provide evidence that the serine protease activity of the Nudel protein is essential for embryonic dorsoventral polarity and that the active Nudel protease is generated by autoproteolytic cleavage of a zymogen form. Activation of the Nudel protease is independent of the other known proteases involved in dorsoventral polarity establishment and appears to occur symmetrically on the surface of the embryo. Our findings suggest that Nudel protease activation initiates the protease cascade that produces the ventral signal, but that spatial regulation occurring downstream of Nudel protease activation localizes the cascade to the ventral side of the embryo. PMID- 9735366 TI - The enhancer of polycomb gene of Drosophila encodes a chromatin protein conserved in yeast and mammals. AB - The Polycomb group of genes in Drosophila are homeotic switch gene regulators that maintain homeotic gene repression through a possible chromatin regulatory mechanism. The Enhancer of Polycomb (E(Pc)) gene of Drosophila is an unusual member of the Polycomb group. Most PcG genes have homeotic phenotypes and are required for repression of homeotic loci, but mutations in E(Pc) exhibit no homeotic transformations and have only a very weak effect on expression of Abd-B. However, mutations in E(Pc) are strong enhancers of mutations in many Polycomb group genes and are also strong suppressors of position-effect variegation, suggesting that E(Pc) may have a wider role in chromatin formation or gene regulation than other Polycomb group genes. E(Pc) was cloned by transposon tagging, and encodes a novel 2023 amino acid protein with regions enriched in glutamine, alanine and asparagine. E(Pc) is expressed ubiquitously in Drosophila embryogenesis. E(Pc) is a chromatin protein, binding to polytene chromosomes at about 100 sites, including the Antennapedia but not the Bithorax complex, 29% of which are shared with Polycomb-binding sites. Surprisingly, E(Pc) was not detected in the heterochromatic chromocenter. This result suggests that E(Pc) has a functional rather than structural role in heterochromatin formation and argues against the heterochromatin model for PcG function. Using homology cloning techniques, we identified a mouse homologue of E(Pc), termed Epc1, a yeast protein that we name EPL1, and as well as additional ESTs from Caenorhabditis elegans, mice and humans. Epc1 shares a long, highly conserved domain in its amino terminus with E(Pc) that is also conserved in yeast, C. elegans and humans. The occurrence of E(Pc) across such divergent species is unusual for both PcG proteins and for suppressors of position-effect variegation, and suggests that E(Pc) has an important role in the regulation of chromatin structure in eukaryotes. PMID- 9735367 TI - Production and activity of spore differentiation factors (SDFs) in Dictyostelium. AB - SDF-1 and SDF-2 are peptides that promote terminal spore differentiation under submerged conditions. The present study shows that they accumulate differentially and are released during the development of wild-type cells and can promote spore formation in cells disaggregated from wild-type culminants. SDF-1 accumulates during the slug stage and is released in a single burst at the onset of culmination while SDF-2 accumulates during early culmination and is released in a single burst from mid-culminants. The effects of SDF-1 and SDF-2 on stalk cell formation in cell monolayers were investigated. SDF-1 by itself induces stalk cell formation in some strains and also synergizes with the stalk-cell-inducing factor, DIF-1. cAMP has an inhibitory effect on stalk cell formation when either DIF-1 or SDF-1 are present on their own but is almost not inhibitory when both are present. SDF-2 alone does not induce stalk cell formation and appears to inhibit the response to DIF-1. At the same time, it increases the extent of vacuolization of the stalk cells that are produced. We propose that the release of SDF-1 and then of SDF-2 may mark irreversible steps in the developmental programme associated, respectively, with culmination and spore maturation. PMID- 9735368 TI - The K box, a conserved 3' UTR sequence motif, negatively regulates accumulation of enhancer of split complex transcripts. AB - Cell-cell interactions mediated by the Notch receptor play an essential role in the development of the Drosophila adult peripheral nervous system (PNS). Transcriptional activation of multiple genes of the Enhancer of split Complex [E(spl)-C] is a key intracellular response to Notch receptor activity. Here we report that most E(spl)-C genes contain a novel sequence motif, the K box (TGTGAT), in their 3' untranslated regions (3' UTRs). We present three lines of evidence that demonstrate the importance of this element in the post transcriptional regulation of E(spl)-C genes. First, K box sequences are specifically conserved in the orthologs of two structurally distinct E(spl)-C genes (m4 and m8) from a distantly related Drosophila species. Second, the wild type m8 3' UTR strongly reduces accumulation of heterologous transcripts in vivo, an activity that requires its K box sequences. Finally, m8 genomic DNA transgenes lacking these motifs cause mild gain-of-function PNS defects and can partially phenocopy the genetic interaction of E(spl)D with Notchspl. Although E(spl)-C genes are expressed in temporally and spatially specific patterns, we find that K box-mediated regulation is ubiquitous, implying that other targets of this activity may exist. In support of this, we present sequence analyses that implicate genes of the iroquois Complex (Iro-C) and engrailed as additional targets of K box-mediated regulation. PMID- 9735370 TI - Injection of sperm extract mimics spatiotemporal dynamics of Ca2+ responses and progression of meiosis at fertilization of ascidian oocytes. AB - Sperm extract (SE) of the ascidian, Ciona savignyi, injected into oocytes induced repetitive intracellular Ca2+ increases with kinetics consistent with those at fertilization and caused reinitiation and progression of meiosis as in fertilized oocytes with the formation of polar bodies. The Ca2+ response comprised two sets of Ca2+ oscillations separated by 5 minutes and correlated with the first and second meiotic metaphase. The effects of SE were dose dependent and the critical dose corresponded roughly to a single spermatozoon. In the first Ca2+ transient observed by confocal microscopy, a Ca2+ wave started from the SE injection site at the peripheral region of the oocyte and propagated across the ooplasm. The similar wave was produced by injection at the central region, starting from an arbitrary cortical area after 30 seconds, probably after SE had diffused to the cortex. The sensitivity to SE is thought to be preferentially higher in the cortex. The effective component of SE was heat-unstable, and its molecular weight was estimated as in the range between 10x10(4 )and 3x10(4) using membrane filters. These results suggest that, in ascidian fertilization, a cytosolic sperm protein factor is introduced to the oocyte cortex and induces Ca2+ waves and thereby meiotic resumption, leading to cell-cycle-correlated Ca2+ oscillations. PMID- 9735369 TI - miranda localizes staufen and prospero asymmetrically in mitotic neuroblasts and epithelial cells in early Drosophila embryogenesis. AB - When neuroblasts divide, prospero protein and mRNA segregate asymmetrically into the daughter neuroblast and sibling ganglion mother cell. miranda is known to localize prospero protein to the basal cell cortex of neuroblasts while the staufen RNA-binding protein mediates prospero mRNA localization. Here we show that miranda is required for asymmetric staufen localization in neuroblasts. Analyses using miranda mutants reveal that prospero and staufen interact with miranda under the same cell-cycle-dependent control. miranda thus acts to partition both prospero protein and mRNA. Furthermore, miranda localizes prospero and staufen to the basolateral cortex in dividing epithelial cells, which express the three proteins prior to neurogenesis. Our observations suggest that the epithelial cell and neuroblast (both of epithelial origin) share the same molecular machinery for creating cellular asymmetry. PMID- 9735371 TI - Regulation of touch receptor differentiation by the Caenorhabditis elegans mec-3 and unc-86 genes. AB - The nematode Caenorhabditis elegans possesses six morphologically similar neurons that are responsible for sensing gentle touch to the body. Previous genetic studies identified genes that are necessary for the production and differentiation of these touch cells. In particular, unc-86 encodes a POU-type homeodomain protein needed for the production of the touch cells, while mec-3 encodes a LIM-type homeodomain protein needed for the differentiation of the touch cells. Molecular studies showed that MEC-3 and UNC-86 bind cooperatively to sites in the mec-3 promoter and can synergistically activate transcription from it in vitro. Here we show that UNC-86::MEC-3 hetero-oligomer-binding sites are also found in the promoters of two presumed targets of mec-3, the mec-4 and mec-7 genes, that are necessary for the function of the touch cells. These sites, which are well-conserved in the related nematode C. briggsae, are required for promoter activity. When one of the binding sites is cloned into a heterologous promoter, expression is found in the touch cells and two to four other cells that express mec-3 and unc-86. These data support a model in which touch-cell differentiation is specified, in part, by the UNC-86::MEC-3 hetero-oligomer and not by MEC-3 alone. Ectopic expression of mec-3, driven by a heat-shock promoter, also supports this hypothesis: the acquisition of touch-cell characteristics by several additional cells under these conditions required unc-86. Since the touch cell lineages express UNC-86 before MEC-3, MEC-3 appears to modify the activity of UNC-86, leading to touch-cell-specific gene expression. Because both UNC-86 and MEC-3 have activation domains, the formation of the hetero-oligomer may create a strong activator. In the modification of UNC-86 function by MEC-3 in the touch cells, these studies provide an example of how the sequential activation of transcription factors can determine cell fate within particular cell lineages. PMID- 9735372 TI - Tyrosine kinase inhibition produces specific alterations in axon guidance in the grasshopper embryo. AB - Tyrosine kinase signaling pathways are essential for process outgrowth and guidance during nervous system development. We have examined the roles of tyrosine kinase activity in programming growth cone guidance decisions in an intact nervous system in which neurons can be individually identified. We applied the tyrosine kinase inhibitors herbimycin A and genistein to whole 40% grasshopper embryos placed in medium, or injected the inhibitors into intact grasshopper eggs. Both inhibitors caused interneuronal axons that normally would grow along the longitudinal connectives to instead leave the central nervous system (CNS) within the segmental nerve root and grow out toward the body wall muscles. In addition, herbimycin A produced pathfinding errors in which many longitudinal axons crossed the CNS midline. To study how this drug affected guidance decisions made by individual growth cones, we dye-filled the pCC interneuron, which normally extends an axon anteriorly along the ipsilateral longitudinal connective. In the presence of herbimycin A, the pCC growth cone was redirected across the anterior commissure. These phenotypes suggest that tyrosine kinase inhibition blocks a signaling mechanism that repels the growth cones of longitudinal connective neurons and prevents them from crossing the midline. PMID- 9735373 TI - The role of the SMN gene in proximal spinal muscular atrophy. AB - Childhood spinal muscular atrophy (SMA) is a common recessive autosomal disorder that results in degeneration of lower motor neurons. The identification of the disease gene, Survival of Motor Neuron (SMN), was a major advance in understanding the molecular basis underlying this devastating neuromuscular disease. This finding has greatly improved the genetic counselling of SMA families. Recently, biochemical studies demonstrated its involvement in the biogenesis of spliceosomal snRNPs, suggesting a critical role of SMN in RNA processing. Surprisingly, other studies showed a putative role of SMN in an anti apoptotic pathway involving Bcl-2. The function of SMN protein is not fully understood. These observations emphasized the difficulty in elucidating the function of any novel protein. Therefore, multidisciplinary approaches are required to understand the pathogenesis of SMA. PMID- 9735374 TI - The genetics of psoriasis: a complex disorder of the skin and immune system. AB - In the last few years, molecular genetics analyses have permitted novel insights into psoriasis, a disease characterized by uncontrolled proliferation of keratinocytes and recruitment of T cells into the skin. The disease affects approximately 1-2% of the Caucasian population and can occur in association with other inflammatory diseases such as Crohn's disease and in association with human immunodeficiency virus (HIV) infection. Given that psoriasis has characteristics of an autoimmune disease, it is not surprising that HLA studies revealed an association with certain alleles, notably HLA-Cw6. Despite this HLA component, psoriasis in some families is inherited as an autosomal dominant trait with high penetrance. Loci at chromosome 17q25 and 4q have been identified following genome wide linkage scans of large, multiply affected families. In the case of at least the susceptibility locus at 17q25, the development of psoriasis does not require the presence of HLA-Cw6. Sib-pair analyses have confirmed the association with HLA-Cw6, confirmed the existence of a locus at 17q25 and identified other possible susceptibility loci. Two independent groups have reported a third region on chromosome 20p. Despite these findings, the extent of genetic heterogeneity and the role of environmental triggers and modifier genes is still not clear. The precise role of HLA also still needs to be defined. The isolation of novel susceptibility genes will provide insights into the precise biochemical pathways that control this disease. Such pathways will also reveal additional candidate genes that can be tested for molecular alterations resulting in disease susceptibility. PMID- 9735375 TI - Gene defect behind APECED: a new clue to autoimmunity. AB - The molecular background of human autoimmunity is poorly understood. Although many autoimmune diseases have a genetic basis, the actual disease appearance results from a complex interplay between genes and environment and thus these diseases represent typical multifactorial diseases. Even with molecular tools provided by the Human Genome Project, it still remains a challenge to identify the predisposing DNA variants behind such multifactorial traits. Two strategies have been suggested to provide short-cuts to the dissection of the genetic background of complex autoimmune diseases: (i) identification of genes in rare human diseases with a strong autoimmune component or (ii) unravelling loci causing phenotypes resembling autoimmune diseases in inbred mice strains. Autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED) is a monogenic autosomal disease with a recessive inheritance pattern, characterized by multiple autoimmune endocrinopathies, chronic mucocutaneous candidiasis and ectodermal dystrophies. Since it is the only known human autoimmune disease inherited in a Mendelian fashion, it provides an excellent model to analyse the genetic component of human autoimmunity. The causative gene for APECED was isolated recently by a traditional positional cloning strategy by two independent groups. The cDNA for the APECED gene proved to originate from a novel gene, AIRE , which is expressed prevalently in thymus, pancreas and adrenal cortex. Multiple mutations in AIRE have been identified in APECED patients. The predicted proline rich AIRE polypeptide harbours two PHD-type zinc finger motifs and contains a putative nuclear targeting signal suggesting its involvement in the regulation of transcription. In the future, functional analysis of the AIRE protein both in vitro and in vivo will provide valuable insight not only into the molecular pathogenesis of APECED but also into the aetiology of autoimmunity in general. PMID- 9735376 TI - ATM: from gene to function. AB - The identification of ATM , the gene responsible for the pleiotropic recessive disease ataxia telangiectasia, has initiated extensive research to determine the functions of its multifaceted protein product. The ATM protein belongs to a family of protein kinases that share similarities at their C-terminal region with the catalytic domain of phosphatidylinositol 3-kinases. Studies with ataxia telangiectasia (A-T) cells and Atm-deficient mice have shown that ATM is a key regulator of multiple signaling cascades which respond to DNA strand breaks induced by damaging agents or by normal processes, such as meiotic or V(D)J recombination. These responses involve the activation of cell cycle checkpoints, DNA repair and apoptosis. Other roles outside the cell nucleus might be carried out by the cytoplasmic fraction of ATM. In addition, ATM appears to function as a 'caretaker', suppressing tumorigenesis in specific T cell lineages. PMID- 9735377 TI - Two rights make a wrong: human left-right malformations. AB - Like all vertebrates, humans establish anatomical left-right asymmetry during embryogenesis. Variation from this normal arrangement (situs solitus) results in heterotaxy, expressed either as randomization (situs ambiguus) or complete reversal (situs inversus) of normal organ position. Familial heterotaxy occurs with autosomal dominant, recessive and X-linked inheritance. All possible situs variants, solitus, ambiguus and inversus, can appear among some heterotaxy families. Positional cloning has led to the identification of a gene on the X chromosome responsible for some cases of human heterotaxy. Additional candidate genes have emerged from recent studies of left-right axis development in chick, frog and mouse, which have begun to elucidate a tightly regulated genetic cascade that differentiates the left and right sides prior to the appearance of morphological asymmetry. PMID- 9735378 TI - Nuclear genes of human complex I of the mitochondrial electron transport chain: state of the art. AB - The mitochondrial electron transport chain (mtETC) consists of four multi-subunit enzyme complexes. Complex I or NADH:ubiquinone oxidoreductase, the largest mtETC multisubunit complex, consists of approximately 41 subunits. Seven of these subunits are encoded by the mitochondrial genome, the remainder by the nuclear genome. Among the mitochondriocytopathies, complex I deficiencies are encountered frequently. Although some complex I deficiencies have been associated with mitochondrial DNA mutations, the genetic defect has not been elucidated in the majority of complex I-deficient patients. It is expected that many of these patients have mutations in the nuclear-encoded subunits of this complex, so vital for cellular energy production. After a brief summary of the current knowledge of complex I from cow, bacteria and fungi, this review presents the state of the art of the knowledge of the human nuclear-encoded complex I genes which, in the last 18 months, has made enormous progress. At present, the complete gene structure of four subunits and the cDNA structure of 18 of the 34 complex I nuclear-encoded subunits are known. Mapping of these subunits shows a random distribution over the chromosomes. The chromosomal localization is known for 14 complex I genes. Recently, the first mutation, a 5 bp duplication in the 18 kDa (AQDQ) subunit, has been reported. We expect that within 1 year all human nuclear-encoded complex I subunits will be cloned. Mutational analysis of these subunits is warranted in complex I-deficient patients and will not only be important for genetic counselling but will also extend the knowledge regarding the functional properties of the individual human complex I subunits. PMID- 9735379 TI - The hereditary periodic fever syndromes: molecular analysis of a new family of inflammatory diseases. AB - The hereditary periodic fever syndromes are a group of Mendelian disorders characterized by episodic fever and serosal or synovial inflammation. Familial Mediterranean fever (FMF) and the hyperimmunoglobulinemia D and periodic fever syndrome are both recessively inherited, while three dominantly inherited syndromes have been described, the best-characterized of which is familial Hibernian fever (FHF). The last year has seen two major developments in this field: the FMF gene was identified on chromosome 16p by positional cloning, and a second major periodic fever locus was mapped to distal chromosome 12p. The FMF gene (MEFV) encodes a novel 781 amino acid protein; to date, eight different missense mutations and a number of polymorphisms have been described. Seven of the eight mutations occur within a region of 82 amino acids near the C-terminus. Computational analysis of the conceptual protein reveals five different domains/motifs compatible with a nuclear effector function. MEFV is expressed preferentially in granulocytes and myeloid bone marrow precursors, giving rise to speculation that the protein may serve as a transcriptional regulator of inflammation in granulocytes. The second periodic fever locus was mapped by two different groups: one studying FHF, the other studying a similar dominantly inherited syndrome designated familial periodic fever. Both genes map to the same 19 cM region on distal chromosome 12p, strongly suggesting a common locus. The molecular characterization of the periodic fever genes should provide important new insights into the regulation of inflammation in general. PMID- 9735380 TI - The fundamental and medical impacts of recent progress in research on hereditary hearing loss. AB - What would define real progress in the field of deafness research in fundamental and medical terms? In fundamental terms, progress would be measured by an improvement in our knowledge of the development and physiology of the ear. In medical terms, progress would lead to the division of the broad category of hearing defects into distinct clinical entities or subclasses, the collection of epidemiological data, the creation of molecular diagnostic tests, the improvement of genetic counselling services and the development of new therapeutics. In this review, we will introduce some general considerations on hereditary hearing loss and on the structure and function of the ear, present the rapidly emerging data on the molecular basis of syndromic and non-syndromic forms of hearing loss and comment on relevant recent progress in this field of research. Generally speaking, the isolation of genes underlying hereditary hearing loss has, as yet, had little impact on our understanding of the biology of the ear, whereas it has made major contributions to the medical field, in particular due to the recognition of two genes, Cx26 and mitochondrial 12S rRNA , as frequently underlying cases of non-syndromic hearing impairment. PMID- 9735381 TI - A catalogue of imprinted genes and parent-of-origin effects in humans and animals. AB - Parent-of-origin effects were first recorded >3000 years ago by mule breeders in Asia Minor. There are now several different types of evidence suggesting the presence of a large number of imprinted genes, many of which have not yet been identified. Here, we catalogue a wide range of evidence and phenomena which indicate or suggest the presence of genomic imprinting in animals. This evidence includes: the direct documentation of parent-of-origin-specific gene transcription; human disease inheritance patterns which suggest the involvement of imprinted genes; and older, less well studied animal models which may show parent-of-origin effects. PMID- 9735382 TI - Position effect in human genetic disease. AB - The spatially, temporally and quantitatively correct expression of a gene requires the presence not only of intact coding sequence, free of adverse nucleotide changes, but also correctly functioning regulatory control. With the identification of an increasing number of disease-related genes, the molecular defect in many cases has been defined. It is becoming clear that it is not always the transcription unit that bears the defect: there are a number of cases where the regulation of gene expression has been compromised. Cases associated with chromosomal rearrangement outside the transcription and promoter regions are categorized as position effects. A number of different mechanisms may explain their aetiology. Here, we examine the human disorders where such position effects are implicated. Further study of such cases may lead to important insights into mechanisms of gene regulation and transcriptional control. PMID- 9735383 TI - Chromosome painting: a useful art. AB - Chromosome 'painting' refers to the hybridization of fluorescently labeled chromosome-specific, composite probe pools to cytological preparations. Chromosome painting allows the visualization of individual chromosomes in metaphase or interphase cells and the identification of both numerical and structural chromosomal aberrations in human pathology with high sensitivity and specificity. In addition to human chromosome-specific probe pools, painting probes have become available for an increasing range of different species. They can be applied to cross-species comparisons as well as to the study of chromosomal rearrangements in animal models of human diseases. The simultaneous hybridization of multiple chromosome painting probes, each tagged with a specific fluorochrome or fluorochrome combination, has resulted in the differential color display of human (and mouse) chromosomes, i.e. color karyotyping. In this review, we will summarize recent developments of multicolor chromosome painting, describe applications in basic chromosome research and cytogenetic diagnostics, and discuss limitations and future directions. PMID- 9735384 TI - Mouse mutagenesis-systematic studies of mammalian gene function. AB - The mouse will play a pivotal role in mammalian gene function studies as we enter the post-genomics era. The challenge is to develop systematic, genome-wide mutagenesis approaches to the study of gene function. The current mouse mutant resource has been an important source of human genetic disease models. However, despite an apparently large catalogue of mouse mutations, we have access to mutations at only a small fraction of the likely total number of mammalian genes there is a phenotype gap that needs to be filled by the establishment of new mutagenesis programmes. Two routes, genotype- and phenotype-driven, can be used for the recovery of novel mouse mutations. For the former, gene trap embryonic stem cell libraries appear set to deliver a large number of mutations around the mouse genome. The advantage of genotype-driven approaches is the ease of identification of the mutated locus; the disadvantage that a priori assumptions have to be made concerning the function and likely phenotype of the mutated gene. In contrast, phenotype-driven mutagenesis emphasizes the recovery of novel phenotypes. One phenotype-driven approach that will play an important role in expanding the mouse mutant resource employs the mutagen N-ethyl-N-nitrosourea (ENU). The phenotype-driven route makes no assumptions about the underlying genes involved, and ENU mutagenesis programmes can be expected to play a significant role in uncovering novel pathways and genes; the disadvantage is that the identification of the mutant gene is still not trivial. Together, the complementary routes of genotype- and phenotype-driven mutagenesis will provide a much enlarged catalogue of mouse mutations and phenotypes for future gene function studies. PMID- 9735385 TI - Engineering mammalian chromosomes. AB - Construction of a mammalian artificial chromosome (MAC) will develop our understanding of the requirements for normal chromosome maintenance, replication and segregation while offering the capacity for introducing genes into cells. Construction of MACs with telomere, centromere and replication function has been approached by two methods. The 'top down' strategy uses artificially induced chromosome truncations as a means to define a minimal chromosome that retains the mitotic properties of a normal chromosome. The 'build up' approach has focused on attempts to assemble MAC vectors containing functionally defined telomere repeats together with candidate centromere and replication origin sequences. Here we report on significant advances in both areas, with particular emphasis on two reports showing that stable, low copy number MACs containing a functional centromere can be produced following transfection of naked DNA into the human HT1080 cell line. One approach used a transfection mixture of cloned synthetic alpha-satellite arrays up to 1 Mb in length and unlinked telomeric DNA, in either the presence or absence of random human genomic DNA fragments. In the second approach, MACs were formed from a defined yeast artificial chromosome (YAC) DNA molecule containing 100 kb of highly homo- geneous alphoid DNA retrofitted with human telomere repeats. These results demonstrate for the first time that alpha satellite DNA can seed de novo centromeres in human cells, indicating that this repetitive sequence family plays an important role in centromere function. The stability of these MACs suggests that they have potential to be developed as gene delivery vectors. PMID- 9735386 TI - A practical guide to orient yourself in the labyrinth of genome databases. AB - The identification of genes involved in human inherited disorders has been revolutionized by the resources produced by the Human Genome Project. In particular, the generation of >1 000 000 human expressed sequence tags (ESTs) has led to the partial identification of a significant percentage of all human genes. In the next 7 years, we will witness another revolution when sequencing of the human genome is complete. The generation of large amounts of genomic data must be accompanied by parallel efforts to make the information easily accessible. Efforts towards this goal have already started, but retrieval of information from genomic databases still remains an arduous task. With practical examples, we will try to show how the currently available information can be exploited usefully, in particular to identify candidate genes for human diseases. PMID- 9735387 TI - Ribozymes as therapeutic tools for genetic disease. AB - The discovery that RNA can act as a biological catalyst, as well as a genetic molecule, indicated that there was a time when biological reactions were catalysed in the absence of protein-based enzymes. It also provided the platform to develop those catalytic RNA molecules, called ribozymes, as trans -acting tools for RNA manipulation. Viral diseases or diseases due to genetic lesions could be targeted therapeutically through ribozymes, provided that the sequence of the genetic information involved in the disease is known. The hammerhead ribozyme, one of the smallest ribozymes identified, is able to induce site specific cleavage of RNA, with ribozyme and substrate being two different oligoribonucleotides with regions of complementarity. Its ability to down regulate gene expression through RNA cleavage makes the hammerhead ribozyme a candidate for genetic therapy. This could be particularly useful for dominant genetic diseases by down-regulating the expression of mutant alleles. The group I intron ribozyme, on the other hand, is capable of site-specific RNA trans splicing. It can be engineered to replace part of an RNA with sequence attached to its 3' end. Such application may have importance in the repair of mutant mRNA molecules giving rise to genetic diseases. However, to achieve successful ribozyme-mediated RNA-directed therapy, several parameters including ribozyme stability, activity and efficient delivery must be considered. Ribozymes are promising genetic therapy agents and should, in the future, play an important role in designing strategies for the therapy of genetic diseases. PMID- 9735388 TI - The therapeutic reactivation of fetal haemoglobin. AB - Unusually high levels of fetal haemoglobin production can ameliorate sickle cell disease and beta thalassaemia. Although efforts directed at the pharmacological stimulation of fetal haemoglobin as an approach to managing these conditions have met with limited success, there is wide variation in individual responses. Whether this reflects the particular mutations that underlie these conditions or other genetic factors remains to be determined, as does the ideal combination of agents to achieve this end. These results are encouraging, however, in particular in view of the recent demonstration that other monogenic diseases, Duchenne muscular dystrophy, for example, might be amenable to the same therapeutic strategy. PMID- 9735389 TI - Enhanced in vitro cytotoxicity and cytostasis of the combination of onconase with a proteasome inhibitor. AB - In proliferating cells the turnover rate of proteins responsible for regulation of the cell cycle progression, namely cyclins and inhibitors of the cyclin dependent kinases (CDKs) and phosphatases, is rapid and their cellular level is modulated at the transcriptional, translational and/or degradation (via proteasome pathway) stages. Inhibition of proteasome function results in accumulation of rapidly turning over proteins and, thus, causes an imbalance of the cell cycle regulatory components, and loss of their regulatory function. Indeed, it has been shown that proteasome inhibitors perturb the cell cycle progression. Onconase, a novel RNase which has anti-tumor activity and is in clinical trials, has previously been shown to suppress protein synthesis, presumably by degradation of intracellular RNA, preferentially tRNA. By interfering with regulation of expression of cyclins and/or CDK-inhibitors, onconase also may induce the imbalance of these proteins and potentiate the effect of proteasome inhibitors. In the present study, we observed that the combinations of onconase with peptide-aldehyde inhibitors of calpain and proteasome such as the N-acetyl-leucinyl-leucinyl-norleucinal (LLnL) and the N acetyl-leucinyl-valinyl-phenylalaninal (LVP), but not N-acetyl-leucinyl-leucinyl methioninal (LLM), were synergistic in suppressing cell proliferation and inducing apoptosis in three human tumor cell lines: A-549 lung adenocarcinoma, DU 145 prostatic carcinoma, and MDA-MB-231 breast carcinoma. The observed cytotoxicity may also be a result of prevention of the induction of the 'survival' genes by the nuclear factor kappaB (NFkappaB) by onconase and proteasome inhibitors. The data indicate that such combinations should be further tested as potential anti-cancer regimens. PMID- 9735390 TI - Thrombomodulin, a receptor for the serine protease thrombin, is decreased in primary tumors and metastases but increased in ascitic fluids of patients with advanced ovarian cancer FIGO IIIc. AB - The human ovarian cancer cell line OV-MZ-19, established from a patient with cystadenocarcinoma of the ovary, expressing thrombomodulin (TM), a cell surface receptor for the serine protease thrombin, interacts with monoclonal and polyclonal antibodies having different specificity for TM. These antibodies detect TM antigen by means of flow cytofluorometry, laser scanning microscopy, immunocytochemistry, and ELISA. Therefore a highly sensitive ELISA for TM antigen was established using two different monoclonal antibodies to quantify TM in tissue extracts and biological fluids, e.g. peritoneal malignant ascites. Primary malignant ovarian tumors and metastases of the omentum and intestine contain TM antigen as determined by ELISA but in significantly lower concentrations than benign ovarian tumors (p=0.0056). In contrast, malignant ascitic fluid of patients with advanced ovarian cancer (FIGO IIIc) contain significantly elevated concentrations of soluble TM than benign peritoneal exudates (p=0.0003). Immunoaffinity purified ascites-derived TM efficiently activates protein C. Protein C activation of ascites-derived TM as well as TM expressed by the tumor cells is inhibited by the monoclonal antibodies. TM abrogates the procoagulant activity of thrombin, reduces pericellular thrombin via internalization, accelerates the thrombin-mediated inactivation of pro-uPA, and the EGF domains of TM exhibit mitogenic activity towards fibroblasts and tumor cells. Both, thrombin and pro-uPA play important roles in tumor invasion and metastasis. Therefore, downregulation and/or release of TM into ascitic fluid may play an important role in the malignant behavior of tumor cells. PMID- 9735391 TI - Effects of pharmacokinetic modulating chemotherapy using oral UFT and continuous venous 5FU infusion on the prognosis of irradiated rectal carcinomas with p53 overexpression. AB - We previously found that patients with irradiated rectal carcinomas with p53 overexpression had poor prognoses after radical resection. In the present study, we attempted to improve the prognosis by the introduction of adjuvant chemotherapy. We administered pharmacokinetic modulating chemotherapy, based on the concept that the benefit of a continuous venous 5-fluorouracil (5FU) infusion can be potentiated by low-dose oral UFT, a combination of 1-(2-tetrahydrofuryl)-5 fluorouracil (tegafur) and uracil at a molar ratio of 1:4. Forty-two of 107 patients examined between January 1992 and December 1997 with an irradiated rectal carcinoma (39%) showed positive immunohistochemical staining for p53. Among them, 14 patients received adjuvant chemotherapy (CT group). The percentage of highly malignant tumors in the CT group was higher than that in the no chemotherapy (NCT) group (n=28). However, the rate of cumulative local recurrence in the CT group was 0%, while that in the NCT group was 28.6% (p=0.0392). The distant recurrence rate in the CT group was also significantly lower than that in the NCT group (7.1% vs. 42.9%, p=0.0376). The cumulative 3-year survival rate was 100% in the CT group and 64.3% in the NCT group (p=0.0245). These results suggested that the antitumor property of 5FU enhanced by pharmacokinetic modulation might have a lethal effect on rectal tumors with a loss of the p53 related apoptosis pathway. These preliminary findings are encouraging for the treatment of rectal cancers with possible poor prognosis. PMID- 9735392 TI - Microtubule-damaging drugs triggered bcl2 phosphorylation-requirement of phosphorylation on both serine-70 and serine-87 residues of bcl2 protein. AB - Specifically anti-microtubule agents such as taxol, vincristine, vinblastine and dolastatin can trigger Bcl2 phosphorylation at G2-M phase of the cell cycle in malignant cells derived from a variety of human cancers. In this study, the status of Bcl2 phosphorylation was investigated in response to more antimicrotubule agents such as colchicine, colcemid or podophyllotoxin. Although these agents are not currently used for cancer therapy, they were able to trigger Bcl2 phosphorylation with simultaneous apoptosis in cancer cells. Previously, by using extensive site-directed mutagenesis studies we determined that mutation of serine-70 to alanine could not completely abrogate taxol induced Bcl2 phosphorylation. Studies reported here clearly indicate that serine-87 residue along with serine-70 of Bcl2 protein are necessary for microtubule damaging drug induced phosphorylation. PMID- 9735393 TI - Novel mutation of the PTEN gene in an Italian Cowden's disease kindred. AB - Cowden disease (CD) is an autosomal dominant multiple hamartoma syndrome with an elevated risk of thyroid and breast cancers. The CD susceptibility gene has recently been identified as the PTEN/MMAC1/TEP1 gene localized at 10q23 and coding for a dual specificity protein phosphatase. We report the mutational analysis of the PTEN gene in one Italian CD kindred. By using the single strand conformation polymorphism technique and subsequent direct DNA sequencing of the polymerase chain reaction product, we identified a novel mutation in the exon 5 of the PTEN gene. A heterozygous germline TGT-TAT transition was detected at the nucleotide 407; this causes the amino acid substitution cys136-tyr136 and the generation of a new NSI I restriction site. This mutation was not detected in the unaffected member of the family thereby indicating that it is causally linked to the disease. We ruled out that this mutation is a polymorphic variant because it was not detected in over 100 chromosomes analyzed. Using reverse trancriptase polymerase chain reaction, we detected the expression of the mutant allele in lymphocytes and pathological tissues from an affected member of the family. PMID- 9735394 TI - Alteration of the CDKN2A gene in pancreatic cancers: Is it a late event in the progression of pancreatic cancer? AB - Various genetic changes are involved in progression of various cancers. We examined alterations (deletion, sequence abnormalities, methylation) of the CDKN2A gene in cell lines and tumor tissues of pancreatic cancers. Some alterations of this gene were found in all the 12 cell lines examined. In the primary lesions of pancreatic cancers, homozygous or hemizygous deletion were found in 8 of 24 ductal carcinoma and 4 of 9 other types of carcinomas. It appears that there is an association between the alteration of this gene and tumor size, regional lymph node metastasis and hematogenous distant metastasis in the ductal carcinoma, but not in the other types of carcinomas. All the 5 liver metastatic lesions of the ductal carcinoma examined revealed homozygous or hemizygous deletion and 3 bp deletion. These results suggest that inactivation of the CDKN2A gene occurs more frequently in cell lines than in pancreatic cancer tissues. Such genetic events on the CDKN2A gene may play an important role possibly at a later step in the progression of pancreatic ductal carcinoma. PMID- 9735395 TI - Usefulness of the high-frequency ultrasound probe in pretherapeutic staging of superficial-type colorectal tumors. AB - We evaluated the usefulness of the high-frequency ultrasound probe (HFUP, 20 MHz) to determine the depth of tumor invasion in 45 patients with superficial colorectal tumors. The correct diagnostic rate was 66% (30/45) when the depth of tumor invasion was classified into the following 6 layers: mucosa (m), upper 1/3 (sm1), middle 1/3 (sm2), and lower 1/3 (sm3) areas of the submucosa, muscularis propria (mp), and the subserosa or deeper areas (s). However, when the depth of tumor invasion was evaluated in 3 layers (m-sm1, sm2-sm3, and mp-deeper layer), which is the classification used to select cases for endoscopic mucosal resection, the correct diagnostic rate was 88.9% (40/45). These results suggest that the HFUP is useful to determine the depth of invasion to select treatment for superficial colorectal tumors. PMID- 9735396 TI - A novel approach for examining the anti-proliferative effect of protein kinase C inhibitors against human astrocytoma cells. AB - Prognosis for astroglial brain tumors that are not amenable to surgical resection remains poor and even successful treatment with current chemoradiotherapy is associated with debilitating sequelae. Consequently, a need to identify novel therapeutics for the treatment of brain tumors remains. Regulation of protein kinase C (PKC) whose activity regulates many cellular functions is crucial for maintaining normal cellular proliferation. Recent reports indicate that malignant glioma cell lines express 100 to 1000-fold higher PKC activity when compared to non-neoplastic astrocytes. In this study we used a novel approach for the evaluation of known PKC inhibitors (CGP 41251, Go 6976, and tamoxifen) as chemotherapeutic agents for the inhibition of growth of an astroglial derived cell line. For this purpose, we constructed a model cell system based on the measurement of light production in cells transfected with the luciferase reporter gene whose expression was quantitated by a highly sensitive, rapid, and easy to perform assay. We isolated U-373MG/MEK1C clone whose highly increased luciferase activity was independent of external stimuli and directly proportional to cell number, and therefore, was used as a measure of cell proliferation to quantitate the effects of several PKC inhibitors on growth of astrocytoma cells in vitro. In conclusion, we have constructed a novel cell system that can be utilized for high throughput screening and identification of potential anti-cancer drugs active against astrocytoma cells in culture. PMID- 9735397 TI - Preclinical antitumor efficacy of S-1: a new oral formulation of 5-fluorouracil on human tumor xenografts. AB - S-1 is a new oral formulation of 5-fluorouracil (5-FU) consisted of 1M tegafur, 0.4M 5-chloro-2,4-dihydroxypyridine that inhibits a degradation of 5-FU, and 1M potassium oxonate that regulates the phosphorylation of 5-FU in the gastrointestinal tract, and has shown excellent antitumor efficacy against various murine tumors in rodents, compared to the oral tegafur-based antitumor drug, UFT (1M tegafur plus 4M uracil), which is used clinically in Japan. To assess the possibility of clinically using S-1, we investigated the antitumor effect of S-1 on various human solid tumor xenografts in athymic rats and mice. In the nude rat system, S-1 was significantly effective against all 12 tumor xenografts tested when its minimum toxic dose (15 mg/kg) was administered for 14 days. Three tumors, stomach (H-81), colon (KM12C) and breast (H-31) markedly regressed in response to treatment with S-1 but not with UFT. The antitumor potency of S-1 was weak against human tumors xenografted into nude mice and likely similar to that of UFT. The reason of the discrepancy in the efficacy of S 1 between rats and mice was found to be that the 5-FU levels in the blood and tumor tissue of rats after oral administration of S-1 persisted much longer than in mice, and this prolonged maintenance of plasma 5-FU levels was significantly related to the potent antitumor activity of S-1. In conclusion, the results of this study suggested that based on its biological and pharmacokinetic characteristics, oral S-1 should be active against various human cancers. PMID- 9735398 TI - Active-MMP2 in cancer cell nests of oral cancer patients: correlation with lymph node metastasis. AB - We examined the gelatinolytic activity in human oral squamous-cell carcinoma tissues in order to evaluate the capability of intravasation and extravasation of cancer cells. By a microdissection-zymography, we demonstrated separately the gelatinolytic activities in cancer cell nests and stroma adjacent to the cancer cells. The gelatinolytic activities, such as pro-matrix metalloproteinase (MMP)9 and active-MMP2 in most of cancer cell nests were much higher than those of normal gingival epithelium. Moreover, the activities of active-MMP2 in cancer cell nests of metastatic cancers were significantly higher than those of non metastatic cancers (p<0.05). These results suggest that active-MMP2 in cancer cells can be a predictive marker for metastasis formation in oral squamous-cell carcinoma patients. PMID- 9735399 TI - Allelic loss on chromosome 22 in oral cancer: possibility of the existence of a tumor suppressor gene on 22q13. AB - In order to understand the detail of genetic alternation on chromosome 22, we performed polymerase chain reaction analysis of microsatellite polymorphisms corresponding to 13 loci on chromosome 22. We examined 33 primary carcinoma tissues, 5 metastatic tissues and corresponding normal tissues. We detected microsatellite instability (MI) in 14 (42.4%) of 33 cases in this study. Loss of heterozygosity (LOH) was observed in at least one locus in 24 (72. 7%) of the 33 cases. Among the loci examined, LOH was restricted to D22S274 on chromosome 22q13 in 11 (40.7%) of 27 informative cases. No significant correlation between histological differentiation and LOH was observed. These observations suggest that the incidence of LOH at chromosome 22q is high and is associated with the carcinogenesis of oral squamous cell carcinoma (SCC). The D22S274 locus may play an important role in the development of oral SCC and be the site harboring a putative tumor suppressor gene. PMID- 9735400 TI - Vanadium induces AP-1- and NFkappB-dependent transcription activity. AB - Vanadate has been reported to be involved in the causation of cancer. In this study, we found that both AP-1 and NFkappaB activities were increased after treatment with sodium vanadate in JB6 cells. Maximum induction of AP-1 and NFkappaB appeared at 48 to 72 h. Phosphorylations of Erks and p38 kinases were markedly increased at 100 microM of vanadate, while phosphorylation of JNKs was not affected. Vanadate also enhances the phosphorylation of IkappaBalpha. These results suggest that the activation of AP-1 and NFkappaB by vanadate may be mediated through enhancement of phosphorylation of Erk/p38 kinases and IkappaBalpha, respectively. PMID- 9735401 TI - Gene amplification as a prognostic factor in primary and secondary high-grade malignant gliomas. AB - The purpose of this study was to examine the incidence of gene amplification in patients with primary (de novo) and secondary high-grade gliomas (gliomas evolving from lower grade malignancies) and to assess its prognostic significance. A total of 186 prospectively collected frozen surgical specimens were analyzed. Extracted DNA was examined by Southern blot using probes corresponding to the EGFR, CDK4, MDM2, n-MYC, CYCD1, PDGFR-alpha, MET, c-MYC oncogenes. Complete clinical data regarding age, sex, tumor size, extent of surgical resection, postoperative therapy and patient survival were collected. We showed that EGFR followed by CDK4 were the most frequent oncogene amplifications. Oncogene amplification events were significantly more frequent in grade 4 than in grade 3 astrocytomas, mixed gliomas or oligodendrogliomas (P<0.001). With respect to EGFR, there was a significant difference in the frequency of amplification between primary and secondary gliomas (P=0.001); however, no difference in the amplification frequency of the other oncogenes was observed. There was no apparent correlation between the occurrence of gene amplification and patient survival, possibly because the genes amplified in human gliomas are part of larger signaling pathways. PMID- 9735402 TI - Construction of phosphorylatable chimeric monoclonal antibody CC49 with a tyrosine srC kinase recognition site. AB - A phosphorylation site for a tyrosine kinase was introduced into chimeric monoclonal antibody CC49 (MAb-chCC49) by inserting a synthetic fragment (Tyr) encoding one tyrosine kinase phosphorylation site into an expression vector. The phosphorylation site was created by incorporating the predicted consensus sequences for phosphorylation by the tyrosine kinase at the carboxyl terminus of the heavy chain constant region of the MAb-chCC49. The resultant modified MAb chCC49 (MAb-chCC49Tyr) was expressed and purified. The MAb-chCC49Tyr protein can be phosphorylated by the tyrosine Src kinase with [gamma-32P]ATP to high radiospecific activity. The 32P-labeled MAb-chCC49Tyr protein binds to cells expressing TAG-72 antigens. The introduction of phosphorylation sites into monoclonal antibodies (MAb) provides a new reagent for the diagnosis and treatment of cancer. This demonstrates that, as was described for the cAMP dependent protein kinase site, a tyrosine phosphorylation site can also be used to introduce phosphorylation sites into proteins. PMID- 9735403 TI - Protein synthesis and transcriptional inhibitors control N-methyl-N'-nitro-N nitrosoguanidine-induced levels of APC mRNA in a p53-dependent manner. AB - In the present study, we show that treatment of wild-type (p53+/+) mouse embryonic fibroblast (MEF) cells with a DNA-alkylating agent, N-methyl-N'-nitro-N nitro-soguanidine (MNNG), resulted in increased levels of adenomatous polyposis coli (APC) mRNA compared to p53 gene-knocked out (p53-/-) MEF cells, indicating that p53 is required for APC expression after alkylation damage. By using HCT-116 colon cancer cells (containing wild-type p53 gene) or p53-/- MEF cells transfected with a pCMV-p53 overexpression plasmid [p53-/-(CMV-p53)], we show that p53 is a labile factor for APC gene expression, and that pretreating HCT-116 cells with a protein synthesis inhibitor, cycloheximide (CHX), inhibited MNNG induced APC mRNA levels by inhibiting p53 protein synthesis. The effect of CHX on p53 protein synthesis was reversible, as the withdrawal of CHX permitted p53 protein synthesis to resume with a concomitant increase in APC mRNA levels after MNNG treatment. To examine whether p53 regulates APC gene expression at the transcriptional level, we treated HCT-116 or p53-/-(CMV-p53) MEF cells with 5,6 dichloro-1-beta-D-ribofuranosylbenzamidazole (DRB; a transcriptional inhibitor), before the MNNG treatment. Although treatment of cells with DRB resulted in increased p53 protein levels, that the APC mRNA levels were decreased suggests that p53 may enhance APC gene expression upstream of the transcriptional machinery where DRB interacts. That the withdrawal of DRB, and subsequent MNNG treatment, increased the level of APC mRNA indicated that the binding of DRB to the transcriptional machinery was reversible. PMID- 9735404 TI - Suppression of heat-induced p53 accumulation and activation by CDDP or x-rays in human glioblastoma cells. AB - We showed that the prominent suppressions of heat-induced accumulation and activation of p53 by CDDP or X-rays were observed in A-172 cells, but not in T98G cells. In addition, the interactive hyperthermic enhancement of CDDP or X-ray cytotoxicity was observed in A-172 cells, but not in T98G cells. Our findings indicate that suppressions of heat-induced accumulation and activation of p53 by CDDP or X-rays contribute positively to an interactive hyperthermic enhancement of CDDP- or X-ray cytotoxicity. PMID- 9735405 TI - DNA binding properties of the nucleocapsid protein from human T-cell leukemia virus type-I. AB - Biochemical and genetic data on retroviral nucleocapsid (NC) proteins have shown that this viral protein exhibits nucleic acid annealing and strand transfer activities and is required for the formation of infectious viral particles. However, the DNA binding properties of the NC protein of the human T-cell leukemia virus type-I (HTLV-I) has not been extensively studied. In this work we characterize the DNA binding ability of the zinc-bound and zinc-free forms of the p15 NC of HTLV-I. We found that only the zinc-bound form of the p15 NC binds single-stranded and double-stranded DNA fragments, but both forms of the p15 NC protein bind and unwind supercoiled DNA. The unwinding activity of the zinc-bound form was 3-fold higher than that observed with the zinc-free form of the protein. Interestingly, eukaryotic DNA topoisomerase antagonists inhibited this unwinding activity. In addition, we showed the formation of NC protein-DNA cleavable complex, which is the result of a presumably covalent bond formed between the protein and the phosphate moiety of the DNA backbone. Moreover, the presence of the p15 NC in the reverse transcription assay significantly increased the activity of the HTLV-I reverse transcriptase. These results demonstrate new DNA binding properties of the p15 NC protein and shed light on the possibility of a novel physiological function for the HTLV-I NC protein in the viral life cycle. PMID- 9735406 TI - Identification of keratins 18, 19 and heat-shock protein 90 beta as candidate substrates of proteolysis during ionizing radiation-induced apoptosis of estrogen receptor negative breast tumor cells. AB - Induction of apoptosis in the estrogen-receptor negative MDA-MB-468 breast tumor cells has been demonstrated by treatments with cytotoxic agents and growth factors. Using these breast tumor cells, we studied ionizing radiation-induced apoptosis. 2D-PAGE of apoptotic cells indicated keratins 18, 19 and heat-shock protein 90 as candidate substrates of apoptosis-associated proteolysis. At the same time, a motif search revealed possible cleavage-sites in keratins 18, 19 (VEVD) and hsp-90 (DEED) that would yield polypeptides of molecular sizes observed experimentally by immunoblotting with specific antisera. This study provides evidence that the insoluble network of intermediate filament proteins of epithelial cells (keratins), and the associated proteins (heat-shock protein 90) constitute targets of caspase-mediated proteolysis during apoptosis triggered by ionizing radiation. PMID- 9735407 TI - Activity of human papillomavirus type 16 P97 promoter in immortal and tumorigenic human oral keratinocytes. AB - We previously immortalized normal human oral keratinocytes (NHOK) by transfection with cloned human papillomavirus type 16 (HPV-16) genome and converted these immortalized cells to tumorigenic cells with chemical carcinogens. Since the tumorigenic cells expressed higher level of HPV-16 E6/E7 transcripts, we predicted that enhanced E6/E7 expression was induced by mutations at the long control region (LCR) of the viral genome integrated into cellular chromosome. To test this possibility, we sequenced the entire HPV-16 LCR from immortalized and tumorigenic cells, but no difference in the sequences in all of the tested cells was observed. However, it is possible that such differences in the expression of E6/E7 could have originated from different activities of cellular transcription factors in the different cells. To examine this prospect, we subcloned entire LCR into a reporter gene and determined the promoter activity of LCR in immortalized and tumorigenic cells. We found that the LCR promoter activity was significantly higher in tumorigenic cells when comparing to immortalized cells. We also observed that at least 477 nucleotides upstream of E6 open reading frame are needed for the maximum LCR promoter activity in tumorigenic cells. PMID- 9735408 TI - Efficacious diagnosis of primary and metastatic human carcinomas with a combination of p53 mutation analysis and clonality analysis of androgen receptor gene. AB - It is occasionally difficult to differentiate between multiple primary and metastatic tumors in case of multiple carcinomas. p53 gene mutation pattern by PCR-SSCP and a clonality analysis on the human androgen receptor gene (HUMARA) in multiple tumors were used to distinguish whether carcinomas were multiple primary tumors or a metastasis. Three female patients with multiple carcinomas, who had undergone surgical resection, were selected for these analyses as typical candidates from 11 cases tested. In the first case with bilateral breast cancer the combined analyses were performed on DNA extracted from tumor portions in paraffin sections of each tumor sample and the results indicated that the carcinomas consisted of a primary carcinoma and metastatic tumors. The carcinomas in the second patient consisted of multiple primary carcinomas and the tumors in the third patient consisted of two independent primary carcinomas with different clonal origins. Thus a combination of p53 gene mutation analysis and clonality analysis on HUMARA provided useful, reliable evidence which can help in discriminating multiple primary tumors from metastatic tumors. PMID- 9735409 TI - Transforming growth factor-beta enhances the ultraviolet-mediated stress response in p53-/- keratinocytes. AB - Skin cancer is the most common tumor type in Caucasians, with an incidence that approaches the lifetime risk for all other cancer subtypes combined. The most common predisposing factor in the development of non-melanoma skin cancer is exposure to ultraviolet (UV) radiation in sun-light. UV radiation activates c-Jun amino-terminal kinases (JNK); this kinase pathway is involved in UV-mediated apoptosis and phosphorylation of c-Jun, all of which are part of the cellular stress response. Transforming growth factor-beta1 (TGF-beta1) is an important negative regulator of keratinocyte proliferation and has other pleiotropic effects in these cells. The purpose of these investigations was to decide whether TGF-beta1 activated c-Jun amino-terminal kinases in a spontaneously immortalized human keratinocyte cell line, HaCaT, and if TGF-beta1 modulated the activation of JNK in keratinocytes exposed to ultraviolet C (UVC) radiation. Results from these investigations showed that TGF-beta1 (10 ng/ml) activated JNK within 5 min. Pretreatment with TGF-beta1 enhanced UV-mediated JNK activation and was time- and UV-dose-dependent. Pretreatment with TGF-beta1 also enhanced activity of the c Jun promoter-reporter construct, TRE(x5)-CAT. These results suggested that TGF beta1 modulates the response of keratinocytes to ultraviolet radiation and implicates TGF-beta1 as a potential mediator the cellular of stress response in keratinocytes. PMID- 9735410 TI - Antitumor activity of human umbilical cord blood cells: A comparative analysis with peripheral blood and bone marrow cells. AB - Although the hematopoietic reconstituting ability of human umbilical cord blood cells (UCBC) is well documented, their antitumor cytotoxic potential has not been well studied. Therefore, UCBC were compared to normal peripheral blood stem cells (PBSC) and bone marrow (BM) stem cell harvests for cytomorphology, antitumor cytotoxic activity before and after ex vivo cytokine manipulation, response to T and B cell mitogens, expression of adhesion molecules and immunophenotypes using flow cytometry, cytokine production and in vivo antitumor activity. BM and PBSC, but not UCBC, did not form cellular clusters in culture. More cytotoxic granules were present in the cytoplasm of UCBC than PBSC following activation in vitro. Ex vivo manipulation of UCBC with cytokines produced more cytotoxicity to K562 and Raji tumor cells than PBSC or BM (p<0.001). Most cytotoxic cells in UCBC cultures were T lymphocytes, and a correlation existed between the number of CD56+ cells and cytotoxicity levels, particularly after in vitro activation with interleukin 2. No significant difference in adhesion molecule expression was noted among UCBC, PBSC and BM cells. However, there was a significantly decreased expression of CD54 molecules (ICAM) on UCBC compared to PBSC (p<0.05). IL-2 activated UCBC showed significant antitumor effects against K562 leukemic cells grown in SCID mice. Thus UCBC contained more antitumor effector cells and precursors than cells from marrow or peripheral blood cells which might be capable of providing a therapeutic effect. PMID- 9735411 TI - A phase II evaluation of pentoxifylline combined with radiation in the treatment of brain metastases. AB - Pentoxifylline (PTX) has pharmacological properties that suggest potential utility as a radiation sensitizer, and preclinical animal studies have been promising. In a non-randomized phase II trial, we used PTX plus standard-dose external-beam whole-brain radiation treatment (WBRT) in patients with brain metastases. Seventeen patients were entered; 14 received both WBRT and PTX and were considered evaluable. Nine of the 14 completed treatment. Analyzing data on all 14 evaluable patients according to intent to treat, median survival time was 33 days, comparable to published data from historical controls. PTX toxicity was not a common cause of patient dropout, supporting higher PTX doses in future trials. PMID- 9735412 TI - Death activation does not stop tumor growth: quantitative evidence for concomitant activation of apoptosis in tumor growth in vitro. AB - A protein-independent fibrosarcoma, Gc-4 PF, grows exponentially in a protein free medium. The doubling time (approximately 26 h) was similar to that of the serum-dependent parental clone, Gc-4 SD cultivated in the presence of fetal calf serum (FCS). We demonstrated here that the protein-free cultivation of Gc-4 PF cells concomitantly activates apoptotic phenotypes (one third of total cell population), including typical morphology, high uptake of Hoechst 33342 dye, and cleavage of DNA to large fragments, as observed in protein-deprived Gc-4 SD cell previously. Gc-4 SD cells arrested in the G0/G1-phase in response to the protein free condition. In contrast, Gc-4 PF cells did not reach G0/G1 arrest in the protein-free condition; instead the durations of both G0/G1 and G2-phases were markedly reduced. The estimation of one cell cycle duration revealed that the cell division cycle was accelerated to 1.7 (27 h/15.4 h)-fold. Then the growth kinetics was able to be verified quantitatively by both the cell division rate and apoptotic cell loss. Protein-free cultivation resulted in slight down regulation of c-myc protein in both cell types, while the down-regulation of p34cdc2, shown clearly in Gc-4 SD cells, was avoided in Gc-4 PF cells. Interestingly, while the expression of p53 was not affected in Gc-4 SD cells in response to the protein-free condition, the suppressor gene product expression was suppressed markedly in Gc-4 PF cells. These results suggest that Gc-4 PF cells may have acquired an ability to accelerate cell division by shortening the cell cycle duration to maintain a proper growth rate in response to intrinsic apoptosis activation with, at least in part, a suppression of p53 expression as well as an escape of down-regulation of p34cdc2. PMID- 9735413 TI - Enhanced antitumour effect of liposomal daunorubicin using antibody-phospholipase C conjugates or fusion protein. AB - We have developed a new two-step method for targeting cytotoxic drugs to tumour cells. The method firstly involves the binding to tumour cells of antibody phospholipase C immunoconjugates or fusion proteins. Further to washing or clearance of the immunoconjugates, liposomes are introduced which are specifically lysed at the tumour site by PLC to release their cytotoxic contents in the vicinity of the tumour cells. For two alternative human cell lines, a synergistic inhibition of cell proliferation was seen for combined treatment with a specific immunoconjugate and daunorubicin encapsulated liposomes. For tumour xenografts in mice, the combined treatment resulted in an inhibition of tumour growth although with no eradication of tumours at the doses used. The two-step antibody-PLC/liposome approach offers broad possibilities for the precise delivery of payloads of cytotoxic drugs to tumour sites. PMID- 9735414 TI - Chromatin, nuclear matrix and the cytoskeleton: role of cell structure in neoplastic transformation (review). AB - Aberrant nuclear and cellular structures are hallmarks of malignant transformation. Thus it is not surprising that the three-dimensional structure of the cell both affects and is affected by changes in gene expression. Here we review the role of the cytoskeleton, nuclear matrix, and chromatin structure in the genesis of cancer. The shape of a cell is governed by a dynamic tissue matrix, which includes extracellular matrix, cytoskeleton and nuclear matrix. Mechanical and chemical signals are transmitted to the nucleus, resulting in alterations in the three-dimensional chromatin organization of genes. The signal transduction pathways affect histone modifications, such as acetylation and phosphorylation, resulting in a relaxed chromatin structure observed in oncogene transformed cells. PMID- 9735415 TI - Bcl-2 overexpression is associated with resistance to paclitaxel, but not gemcitabine, in multiple myeloma cells. AB - Multiple myeloma (MM) is an incurable disease despite an initial response-rate of >60% with conventional or high-dose chemotherapy using glucocorticosteroids (i.e. dexamethasone), or alkylating agents (i.e. melphalan). Although these agents are capable of inducing complete remission (CR) in >50% of MM patients, resistance develops rapidly, in >90% of patients, within 2 years of treatment. Therefore, there is a need for new drugs for the treatment of relapsing and refractory MM patients. Gemcitabine (GEM) is a pyrimidine analog that blocks DNA synthesis, whereas, paclitaxel (TAX) is a mitotic spindle poison that promotes microtubular aggregation. Since it appears that these two drugs have different cellular targets, we examined the effect of each drug individually for several parameters and for possible synergy. We studied the cytotoxic effect of TAX and GEM on MM cells expressing varying levels of the antiapoptotic protein bcl-2, which is overexpressed in the majority of myeloma cell from MM patients. We found that both drugs are cytotoxic by inducing apoptosis, however, the extent of apoptosis with TAX, but not with GEM was dependent on the levels of bcl-2 expression. We further investigated the effect of TAX and GEM on the cell cycle distribution and on the levels of bcl-2. The results indicate that the two drugs have different modes of action with respect to each parameter tested. TAX induced arrest of the cells in the G2/M phase of the cell cycle, regardless of bcl-2 levels, however, apoptosis was induced in mitotic cells expressing relatively low levels of bcl-2. In contrast, GEM caused apoptosis of cells in the S-phase, regardless of level of bcl-2 expression. A major difference between TAX and GEM was in their effects on the levels of bcl-2. Whereas, TAX induced an early downregulation of bcl-2 (only in the cells with relatively low levels of bcl-2), treatment with GEM did not affect bcl-2 levels. The effects of TAX on both the cell cycle and bcl-2 were detected very early (4-8 h) and preceded the onset of apoptosis. GEM and TAX act synergistically, at low doses (IC50 of 0.5 microM for GEM and 0.025 microM for TAX), to effectively kill bcl-2 overexpressing cells that are resistant to higher doses (0.25 microM) of TAX alone. Therefore, we have initiated a phase II clinical trial of TAX and GEM for MM patients refractory to current therapy. PMID- 9735416 TI - Loss of heterozygosity of BRCA1, BRCA2 and ATM genes in sporadic invasive ductal breast carcinoma. AB - The present study was undertaken to analyse the loss of heterozygosity (LOH) of the three genes, BRCA1, BRCA2 and ATM, and their correlation to clinicopathological parameters in sporadic breast cancer. We studied 59 sets of invasive ductal carcinoma, compared to matched normal control DNA. Microsatellite markers intragenic to BRCA1 (D17S1323, D17S1322, D17S855), BRCA2 (D13S1699, D13S1701, D13S1695) and ATM (D11S2179) were simultaneously used. In addition, one marker telomeric to BRCA2 (D13S1694) and four markers flanking ATM were analysed (D11S1816, D11S1819, D11S1294, D11S1818). Thirty-one per cent of the informative cases showed loss of heterozygosity for the BRCA1 gene, 22.8% for BRCA2 gene and 40% for ATM. LOH of BRCA1 correlated with high grade tumors (p=0.0005) and negative hormone receptors (p=0.01). LOH of ATM correlated with higher grade (p=0.03) and a younger age at diagnosis (p=0.03) in our set of tumors. No correlations were detected between BRCA2 LOH and any of the analysed clinicopathological parameters. However, a correlation was detected between allelic loss of the D13S1694 marker, telomeric to BRCA2, and larger tumor sizes and negative estrogen receptors, favoring the hypothesis of the presence of another putative tumor suppressor gene, telomeric to BRCA2, in the 13q12-q14 region. Only 11 tumors had LOH at more than one of the three genes, most of them (6/11) associated LOH of BRCA1 and ATM. One tumor only combined loss of the three genes BRCA1, BRCA2 and ATM. PMID- 9735417 TI - Biomarkers in Barrett's esophagus (review). AB - Patients with Barrett's esophagus where squamous esophageal epithelium has been replaced by columnar epithelium containing goblet cells have an increased risk of esophageal adenocarcinoma. Unfortunately, most of those adenocarcinomas are detected at an advanced stage in which they are incurable. Many biomarkers have been studied, trying to identify the subset of patients who are at very high risk for adenocarcinoma. Dysplasia is at present the for the risk of adenocarcinoma. However, overexpression of p53, allelic losses, proliferation indices, flow cytometric abnormalities, accumulation of acidic fibroblast growth factor, expression of blood group antigens and sucrase-isomaltase during the Barrett's to adenocarcinoma sequence appear promising. However, additional information is needed to justify routine application in clinical practice and to define their role in surveillance programs. PMID- 9735418 TI - Antiproliferative action of tumor necrosis factor-alpha on MCF-7 breastcancer cells is associated with increased insulin-like growth factor binding protein-3 accumulation. AB - Tumor necrosis factor-alpha (TNF-alpha) is a multifunctional cytokine involved in host response to neoplasia. TNF-alpha has been shown to inhibit proliferation and induce apoptosis of MCF-7 breast carcinoma cells. Insulin-like growth factors I and II (IGF-I and IGF-II) are potent mitogens involved in growth regulation of breast epithelial cells and are implicated in the pathophysiology of breast cancer. Their bioactivity is strongly influenced by specific IGF-binding proteins (IGFBPs). We report that accumulation of IGFBP-3 in the conditioned media of MCF 7 cells is increased over control values in the presence of TNF-alpha. The increased IGFBP-3 accumulation induced by TNF-alpha is correlated with increased IGFBP-3 mRNA abundance. TNF-alpha also decreases IGF-I receptor levels in MCF-7 cells. Estradiol-stimulated MCF-7 cell proliferation is associated with reduced IGFBP-3 accumulation, and we show that TNF-alpha attenuation of estradiol stimulated proliferation is associated with increased IGFBP-3 accumulation. Finally, we demonstrate that an IGFBP-3 antisense oligodeoxynucleotide antagonizes TNF-alpha-induced inhibition of cell proliferation and TNF-alpha induced IGFBP-3 accumulation. These data strongly suggest that IGFBP-3 plays a role in modulation of breast cancer cell proliferation by TNF-alpha. PMID- 9735419 TI - Differential transferrin receptor density in human colorectal cancer: A potential probe for diagnosis and therapy. AB - Transferrin receptor density was investigated in human colorectal surgical specimens. Crude membranes were prepared from 23 cancer tumors (adenocarcinoma or malignant villous tumor) and 3 non-cancer tumors (polyadenoma or villous tumor) and 26 adjacent control mucosa. Contrary to non-cancer tumors, Scatchard analysis of 125I-transferrin binding data evidenced higher maximal transferrin binding capacity and lower dissociation constant in cancer tissues (Bmax cancer 1.828+/ 0.320 nmol/g, Kd 24.1+/-4.7 nM), as compared to paired control colonic mucosa (Bmax contol 0.851+/-0.182 nmol/g, Kd 30.7+/-7.3 nM), paired t-tests: Bmax p<0.001, Kd p<0.05). As the cancer/control Bmax ratio was 2.6+/-0.4,transferrin carrier constructs should be proposed for cancer imaging or therapy. PMID- 9735420 TI - Lattice models for proteins reveal multiple folding nuclei for nucleation collapse mechanism. AB - The nature of the nucleation-collapse mechanism in protein folding is probed using 27-mer and 36-mer lattice models. Three different forms for the interaction potentials are used. Three of the four 27-mer sequences have maximally compact and identical native state while the other has a non-compact native conformation. All the sequences fold thermodynamically and kinetically by a two-state process. Analysis of individual trajectories for each sequence using a self-organizing neural net algorithm shows that upon formation of a critical set of contacts the polypeptide chain rapidly reaches the native conformation which is consistent with a nucleation-collapse mechanism. The algorithm, which reduces the identification of the folding nucleus for each trajectory to one of pattern recognition, is used to show that there are multiple folding nuclei. There is a distribution of nucleation contacts in the transition states with some of them occurring with more probability (when averaged over the denatured ensemble) than others. We also show that there is a distribution in the size of the nuclei with the average number of residues in the folding nuclei being less than about one third of the chain size. The fluctuations in the sizes of the nuclei are large, suggestive of a broad transition region. The folding nuclei, the structures of each are the corresponding transition states, have varying degree of overlap with the native conformation. The distribution of the radius of gyration of the transition states shows that these structures are an expanded form (by about 25% in the radius of gyration) of the native conformation. Local contacts are most dominant in the folding nuclei while a certain fraction of non-local contacts is necessary to stabilize the transition states. The search for the critical nuclei initially involves the formation of local contacts, while non-local contacts are formed later. The fractional values of PhiF for the two 27-mer mutants found by using the protein engineering protocol are consistent with the microscopic picture of partial formation of structures involving these residues in the transition state. These observations lead to a multiple folding nuclei (MFN) model for nucleation-collapse mechanism in protein folding. The major implication of the MFN model is that, even if the residues whose tertiary interactions are formed nearly completely in the transition state are mutated, it does not disrupt the nature of the nucleation-collapse mechanism. We analyze the experiments on chymotrypsin inhibitor 2 and alpha-spectrin SH3 domain and two circular permutants in light of the MFN model. It is shown that the PhiF-value analysis for these proteins gives considerable support to the MFN model. The theoretical and experimental studies give a coherent picture of the nucleation-collapse mechanism in which there is a distribution of folding nuclei with some more probable than others. The formation of any specific nucleus is not necessary for efficient two-state folding. PMID- 9735421 TI - SOAP'98. Society for Obstetric Anesthesia and Perinatology, 30th annual meeting. Vancouver, British Columbia, Canada. April 29-May 2, 1998. Abstracts. PMID- 9735422 TI - Large-scale PACS implementation. AB - The transition to filmless radiology is a much more formidable task than making the request for proposal to purchase a (Picture Archiving and Communications System) PACS. The Department of Defense and the Veterans Administration have been pioneers in the transformation of medical diagnostic imaging to the electronic environment. Many civilian sites are expected to implement large-scale PACS in the next five to ten years. This presentation will related the empirical insights gleaned at our institution from a large-scale PACS implementation. Our PACS integration was introduced into a fully operational department (not a new hospital) in which work flow had to continue with minimal impact. Impediments to user acceptance will be addressed. The critical components of this enormous task will be discussed. The topics covered during this session will include issues such as phased implementation, DICOM (digital imaging and communications in medicine) standard-based interaction of devices, hospital information system (HIS)/radiology information system (RIS) interface, user approval, networking, workstation deployment and backup procedures. The presentation will make specific suggestions regarding the implementation team, operating instructions, quality control (QC), training and education. The concept of identifying key functional areas is relevant to transitioning the facility to be entirely on line. Special attention must be paid to specific functional areas such as the operating rooms and trauma rooms where the clinical requirements may not match the PACS capabilities. The printing of films may be necessary for certain circumstances. The integration of teleradiology and remote clinics into a PACS is a salient topic with respect to the overall role of the radiologists providing rapid consultation. A Web-based server allows a clinician to review images and reports on a desk-top (personal) computer and thus reduce the number of dedicated PACS review workstations. This session will focus on effective strategies for a seamless transition. Critical issues involve maintaining a good working relationship with the vendor, cultivating personnel readiness and instituting well-defined support systems. Success depends on the ability to integrate the institutional directives, user expectations and available technologies. A team approach is mandatory for success. PMID- 9735423 TI - Updating a legacy PACS: experience with hardware and operational integration during transition to a new architecture. PMID- 9735424 TI - Enterprise-scale image distribution with a Web PACS. AB - The integration of images with existing and new health care information systems poses a number of challenges in a multi-facility network: image distribution to clinicians; making DICOM image headers consistent across information systems; and integration of teleradiology into PACS. A novel, Web-based enterprise PACS architecture introduced at Massachusetts General Hospital provides a solution. Four AMICAS Web/Intranet Image Servers were installed as the default DICOM destination of 10 digital modalities. A fifth AMICAS receives teleradiology studies via the Internet. Each AMICAS includes: a Java-based interface to the IDXrad radiology information system (RIS), a DICOM autorouter to tape-library archives and to the Agfa PACS, a wavelet image compressor/decompressor that preserves compatibility with DICOM workstations, a Web server to distribute images throughout the enterprise, and an extensible interface which permits links between other HIS and AMICAS. Using wavelet compression and Internet standards as its native formats, AMICAS creates a bridge to the DICOM networks of remote imaging centers via the Internet. This teleradiology capability is integrated into the DICOM network and the PACS thereby eliminating the need for special teleradiology workstations. AMICAS has been installed at MGH since March of 1997. During that time, it has been a reliable component of the evolving digital image distribution system. As a result, the recently renovated neurosurgical ICU will be filmless and use only AMICAS workstations for mission-critical patient care. PMID- 9735425 TI - Computerized follow-up of discrepancies in image interpretation between emergency and radiology departments. AB - Radiographs are ordered and interpreted for immediate clinical decisions 24 hours a day by emergency physicians (EP's). The Joint Commission for Accreditation of Health Care Organizations requires that all these images be reviewed by radiologists and that there be some mechanism for quality improvement (QI) for discrepant readings. There must be a log of discrepancies and documentation of follow up activities, but this alone does not guarantee effective Q.I. Radiologists reviewing images from the previous day and night often must guess at the preliminary interpretation of the EP and whether follow up action is necessary. EP's may remain ignorant of the final reading and falsely assume the initial diagnosis and treatment were correct. Some hospitals use a paper system in which the EP writes a preliminary interpretation on the requisition slip, which will be available when the radiologist dictates the final reading. Some hospitals use a classification of discrepancies based on clinical import and urgency, and communicated to the EP on duty at the time of the official reading, but may not communicate discrepancies to the EP's who initial read the images. Our computerized radiology department and picture archiving and communications system have increased technologist and radiologist productivity, and decreased retakes and lost films. There are fewer face-to-face consultants of radiologists and clinicians, but more communication by telephone and electronic annotation of PACS images. We have integrated the QI process for emergency department (ED) images into the PACS, and gained advantages over the traditional discrepancy log. Requisitions including clinical indications are entered into the Hospital Information System and then appear on the PACS along with images on readings. The initial impression, time of review, and the initials of the EP are available to the radiologist dictating the official report. The radiologist decides if there is a discrepancy, and whether it is category I (potentially serious, needs immediate follow-up), category II (moderate risk, follow-up in one day), or category III (low risk, follow-up in several days). During the working day, the radiologist calls immediately for category I discrepancies. Those noted from the evening, night, or weekend before are called to the EP the next morning. All discrepancies with the preliminary interpretation are communicated to the EP and are kept in a computerized log for review by a radiologist at a weekly ED teaching conference. This system has reduced the need for the radiologist to ask or guess what the impression was in the ED the night before. It has reduced the variability in recording of impressions by EP's, in communication back from radiologists, in the clinical] follow-up made, and in the documentation of the whole QI process. This system ensures that EP's receive notification of their discrepant readings, and provides continuing education to all the EP's on interpreting images on their patients. PMID- 9735426 TI - Prototype Internet consultation system for radiologists. AB - The overall purpose of this study is to develop a prototype radiological consultation system. We concentrate our work on prototype software environment for the system. The system provides a second diagnostic opinion based on similar cases, incorporating the experience of radiologists, their diagnostic rules and a database of previous cases. The system allows a radiologist to enter the description of a particular case using the lexicon such as BI-RADS of American College of Radiology and retrieve the second diagnostic opinion (probable diagnosis) for a given case. The system also allows a radiologist to get other important information too. These advances are based on a new computational intelligence technique and first-order logic. We implemented a rule-based prototype diagnostic system. Two experimental Internet versions are currently available on the web and are under testing and evaluation of design. The diagnosis is based on the opinions of radiologists in combination with the statistically significant diagnostic rules extracted from the available database. PMID- 9735427 TI - The archiving decision: a balance of time, money, and people. PMID- 9735428 TI - Archive selection for the MICAS, a multi-vendor incremental approach to PACS. AB - From the time a decision was made to purchase an archive until a purchase order was issued took approximately 10 months. During this period an RFI was developed, issued and the results analyzed. Technical discussions were held and site visits were made. To ensure that current information was available, a complete review of available multi-modality DICOM compliant archives were made at the 1997 RSNA. With this information in-hand and the future development path for MICAS specified, a detailed RFQ was developed, responses were received and evaluated. A purchase order was to be issued by the end of the first quarter 1998. The archive vendor will have been selected by the time this paper appears in print. The oral presentation of this work will review the responses of the archive vendors and present the basis for selection. It is planned to publish our findings. The archive is the heart and brains of PACS. It controls information acquisition, distribution and storage plus work flow. It is critical that DICOM compliance and interoperability between all components of the PACS be an absolute requirement, especially for the archive. PMID- 9735429 TI - A DICOM document-oriented approach to PACS infrastructure. AB - The need for long-term storage requires the future migration of image data from a PACS to its successor system. This paper considers the cost of such migration It is proposed that storage of data as "documents" in DICOM Part 10 formats on industry-standard media could reduce the time and cost of data migration relative to on-line DICOM transfer. The relation to present efforts in developing document oriented electronic patient records is discussed. DICOM Part 10 files are found to be a sufficient representation of image documents, but additional software tools will be needed to reach its full potential. There is a significant cost benefit of the document storage method, but it is one of many factors which must be balanced in the selection of a PACS. PMID- 9735430 TI - DICOM versus HL7 for modality interfacing. AB - Digital modalities such as CT, MRI, Ultrasound and Computerized Radiography systems, generating softcopy images to be used by a Picture Archiving and Communication System (PACS), need to identify the images properly in order to retrieve and manage them. In many cases, a technologist re-enters patient demographic and study related information at the modality, even although it is usually already present somewhere in the hospital information system (IS). In order to achieve a higher level of efficiency and uniquely identify the created image objects, it is obvious that an interface between the IS and modality to exchange this information is highly desired. There are two options for a modality vendor to implement an IS interface, either using the Health Level (HL7) or Digital Imaging Communication in Medicine (DICOM) communication standard. This paper will explain characteristics of both protocols, and demonstrate that it is preferred to use DICOM versus HL7. In addition, it will show that DICOM is supported by most modality vendors, based on the result of a poll of their Modality Worklist versus HL7 support. PMID- 9735431 TI - A case for automated tape in clinical imaging. AB - Electronic archiving of radiology images over many years will require many terabytes of storage with a need for rapid retrieval of these images. As more large PACS installations are installed and implemented, a data crisis occurs. The ability to store this large amount of data using the traditional method of optical jukeboxes or online disk alone becomes an unworkable solution. The amount of floor space number of optical jukeboxes, and off-line shelf storage required to store the images becomes unmanageable. With the recent advances in tape and tape drives, the use of tape for long term storage of PACS data has become the preferred alternative. A PACS system consisting of a centrally managed system of RAID disk, software and at the heart of the system, tape, presents a solution that for the first time solves the problems of multi-modality high end PACS, non DICOM image, electronic medical record and ADT data storage. This paper will examine the installation of the University of Utah, Department of Radiology PACS system and the integration of automated tape archive. The tape archive is also capable of storing data other than traditional PACS data. The implementation of an automated data archive to serve the many other needs of a large hospital will also be discussed. This will include the integration of a filmless cardiology department and the backup/archival needs of a traditional MIS department. The need for high bandwidth to tape with a large RAID cache will be examined and how with an interface to a RIS pre-fetch engine, tape can be a superior solution to optical platters or other archival solutions. The data management software will be discussed in detail. The performance and cost of RAID disk cache and automated tape compared to a solution that includes optical will be examined. PMID- 9735432 TI - Monotonic noise suppression used to improve the sensitivity of fMRI activation maps. AB - We have introduced a new method of removing noise from images that identifies significant extrema and forces the pixel intensities between any two extrema to change monotonicly. The method has some similarities to wavelet denoising methods we worked with several years ago but is generally more stable and is effective on images with lower SNR's. In this paper the method of monotonic filtering is used to increase the sensitivity in functional magnetic resonance imaging (fMRI) studies. We have used the increased sensitivity to improve the temporal resolution in fMRI studies by roughly a factor of six. A motor activation study was acquired with single slice 256 x 256 pixel T2* weighted images; six cycles of finger tapping were acquired. Each cycle consisted of five images of rest followed by five images of right hand finger tapping followed by five images of left hand finger tapping. The z-scores were calculated and used as the activation map. The left and right activations were both clearly visible when all six cycles were used in the analysis. However, no definitive activation was seen for any one cycle. When the original 256 x 256 images were averaged down to 64 x 64 pixel images before calculation of the z-scores, the activations were partially identified. When the original images were filtered using the monotonic noise reduction algorithm, the left activation was clearly visible in three of the six cycles and partially visible in two others. The right activation was partially visible in 4 out of 6 cycles. Optimized noise reduction should improve the results significantly. The ability to use a single cycles is very important in fMRI studies because many stimuli are more difficult to maintain over many cycles and because complex processes such as in cognitive or memory activity do not have simple responses. PMID- 9735433 TI - Image gallery: a tool for rapid endobronchial lesion detection and display using virtual bronchoscopy. PMID- 9735434 TI - Anatomic labelling of PET brain images with automatic detection of AC and PC. AB - A new automatic method for finding anterior commissure (AC) and posterior commissure (PC) in positron emission tomography(PET) brain image without a reference image is discussed. For labelling and localizing] anatomical structures, a PET image aligned in parallel to the detected AC-PC line is normalized spatially into the corresponding transaxial Talairach brain. This labelling system may provide practical application method to make the clinical evaluation of PET brain images easier. PMID- 9735435 TI - Elastic image registration using correlations. AB - We have developed a multiscale algorithm for elastic registration of images. Rigid registration has many applications but it is often limited by distortions in the images. For example, different views of the same object produce distortions. Common examples of slightly different views producing a distortion can be found in medical imaging, such as matching a current mammogram or chest radiograph with one from a previous year, and in remote sensing, such as matching images taken from different satellite positions. We have developed two methods of elastic registration. Both are multiscale but one used an iterative minimization of the local error and the other uses a windowed correlation. We present preliminary results of the elastic registration method used on windowed correlations. PMID- 9735436 TI - Integration of imaging functionality into the healthcare enterprise using DICOM. AB - The US Department of Veterans Affairs is integrating imaging functionality into the healthcare enterprise using the Digital Imaging and Communication in Medicine (DICOM) standard protocols 1. The VA's VistA Hospital Information System (HIS) is installed at all 170 VA medical centers across the country. Image management is supported by the VistA HIS in several ways. Some VA sites have commercial Picture Archiving and Communication Systems (PACS) interfaced to the VistA HIS, while other sites use the direct image acquisition and diagnostic display capabilities of VistA itself. By supporting a small set of DICOM services, VistA can transmit patient and study text data to the image producing modalities and the commercial PACS, and enable images and study data to be transferred back. Images can be displayed on low-cost clinician's workstations or high-resolution diagnostic quality multi-monitor workstations located within a facility or elsewhere on the healthcare enterprise wide area network. This is a US government work. There are no restrictions on its use. PMID- 9735437 TI - The evolution of electronic imaging in the medical environment. PMID- 9735438 TI - Challenges in the integration of PACS and RIS databases. PMID- 9735439 TI - Experience specifying interface and integration requirements in the PACS request for proposal. PMID- 9735440 TI - Taking the PACS power to the people. PMID- 9735441 TI - MR teleradiology network serving remote imaging centers. AB - The operational experience of a commercial teleradiology practice utilizing a wide-area ISDN network linking six imaging centers located in two states will be reviewed. Open magnet designs were chosen to complement existing high-field units available in each community. Image data was first acquired than transmitted without compression at 128 Kbytes/s to a central reading site located in McLean, Virginia for interpretation by a team of radiologists. Average transmission time was 6-8 minutes. System design allows optimal utilization of radiologists expertise in imaging interpretation while reserving the on-site patient management responsibilities such as gadolinium contrast injections and sedation to a nonradiologists physician and/or nurse practitioner. Over 15,000 teleradiology readings have been rendered via this network by January 1998. PMID- 9735442 TI - An ISDN-based telemedicine system. AB - Our preliminary results show the telemedicine system is successful. It does fulfill the 3 goals we set earlier. However, for such a system to be cost effective, the communication cost need to be cut further. In fact, video is not required at all time. We are hoping to establish criteria and operational procedure for use of video. Also some sort of standard should be set up for evaluation of the quality of telemedicine. PMID- 9735443 TI - Implementing teleradiology in a private radiology practice: lessons learned. AB - The private practice of radiology deals with the delivery of high quality, timely service to referring physicians. Technology offers the promise of supporting this objective through faster reading of studies through electronic imaging and the delivery of results in a fast, efficient manner. In order to be successful, the practice must clearly define its objectives, understand its fundamental business processes and select a technology solution that is proven to meet those objectives. PMID- 9735444 TI - Is PACS a "WAMI" in your institution? It should be! PMID- 9735445 TI - The path to a filmless radiology department: the HUP experience. PMID- 9735446 TI - Continuing quality improvement procedures for a clinical PACS. AB - The University of California at San Francisco (USCF) Department of Radiology currently has a clinically operational picture archiving and communication system (PACS) that is thirty-five percent filmless, with the goal of becoming seventy five percent filmless within the year. The design and implementation of the clinical PACS has been a collaborative effort between an academic research laboratory and a commercial vendor partner. Images are digitally acquired from three computed radiography (CR) scanners, five computed tomography (CT) scanners, five magnetic resonance (MR) imagers, three digital fluoroscopic rooms, an ultrasound mini-PACS and a nuclear medicine mini-PACS. The DICOM (Digital Imaging and Communications in Medicine) standard communications protocol and image format is adhered to throughout the PACS. Images are archived in hierarchical staged fashion, on a RAID (redundant array of inexpensive disks) and on magneto-optical disk jukeboxes. The clinical PACS uses an object-oriented Oracle SQL (systems query language) database, and interfaces to the Radiology Information System using the HL7 (Health Languages 7) standard. Components are networked using a combination of switched and fast ethernet, and ATM (asynchronous transfer mode), all over fiber optics. The wide area network links six UCSF sites in San Francisco. A combination of high and medium resolution dual-monitor display stations have been placed throughout the Department of Radiology, the Emergency Department (ED) and Intensive Care Units (ICU). A continuing quality improvement (CQI) committee has been formed to facilitate the PACS installation and training, workflow modifications, quality assurance and clinical acceptance. This committee includes radiologists at all levels (resident, fellow, attending), radiology technologists, film library personnel, ED and ICU clinician end-users, and PACS team members. The CQI committee has proved vital in the creation of new management procedures, providing a means for user feedback and education, and contributing to the overall acceptance of, and user satisfaction with the system. Well developed CQI procedures have been essential to the successful clinical operation of the PACS as UCSF Radiology moves toward a filmless department. PMID- 9735447 TI - Web technology in the integration of a digital teaching file at the diagnostic workstation. PMID- 9735448 TI - A vascular catheterization simulator for training and treatment planning. AB - We have developed a working prototype of an augmented reality based vascular catheterization simulator. The simulator is called daVinci, and provides the interventional radiologists with a "hands on, user friendly, image based environment to augment training, enhance pretreatment planning and design interventional products and devices. daVinci is based on computational modeling of human anatomical images and provides the user with capabilities for realtime simulated navigation of catheters in both 2-D and 3-D modeled vessels. In addition, multiplaner CT, MRI and anatomical data sets are incorporated into the simulator to enhance the understanding of anatomical relationships associated with vascular catheterization procedures. The current prototype is designed for peripheral vascular applications. Additional systems are currently being considered for both interventional neuroradiology and cardiology. PMID- 9735449 TI - Creating a digital video-based teaching file for interventional procedures using CT fluoroscopy. AB - A computerized radiology education teaching file application, MRW (Multimedia Radiology Workstation), was produced in our department as a collaboration of faculty, fellows and a doctoral student. This inexpensive and flexible system is novice-programmable and is capable of capturing images from multiple modalities (including still and cine image) and organizing them into individual electronic teaching cases. Help and tutorial functions support the main case display functions. PMID- 9735450 TI - Automated routing of DICOM CT, MR, and CR images: solving the pitfalls of vendor specific DICOM implementations. AB - This paper details our experience in developing and implementing an automated DICOM Image Router. This workstation serves as a gateway between image acquisition devices and image display and storage devices. Images are checked for demographic input errors and data format inconsistencies between the source and destination devices. Based on the configured rules, and image may be held for manual correction and/or distributed to multiple locations. Distribution is based on easily configured environmental variables and rules files which may be changed as needed. For example, CT images are typically sent to the archive and to a radiologist's display workstation. If the patient came from the Emergency Department, a copy of the images are sent to a clinician display workstation located in the Emergency Department. If the patient has suffered trauma to the head, a copy of the images are sent to a display workstation in the Neurosurgery Department for possible consultation. The software was developed on a UNIX-based platform and utilizes a Fast Ethernet network. To date, images from a variety of devices have been acquired: General Electric HiSpeed CT/I scanner, General Electric Signa MRI scanner, Philips Thoravision Digital Chest unit, and Fuji AC 3CS Computed Radiography (CR) unit. Each device has presented new challenges in providing a uniform look to patient demographics in the PACS archive. The workstation also provides a buffer in the event of network outages, storing images for later transmission when the network and/or a workstation recover. PMID- 9735451 TI - Providing complete multimedia patient data to consulting radiologists, other specialists, and the referring clinician. PMID- 9735453 TI - Strategies for the promotion of computer applications in radiology in healthcare delivery. AB - The objective of this paper is to identify current trends in the development and implementation of computer applications in today's ever-changing healthcare environment. Marketing strategies are discussed with the goal of promoting computer applications in radiology as a means to advance future healthcare acceptance of technologic developments from the medical imaging field. With the rapid evolution of imaging and and information technologies along with the transition to filmless imaging, radiologists must assume a proactive role in the development and application of these advancements. This expansion can be accomplished in a number of ways including internet based educational programs, research partnerships, and professional membership in societies such as the Society of Computer Applications in Radiology (SCAR). Professional societies such as SCAR, in turn, should reach out to include other professionals from the healthcare community. These would include financial, administrative, and information systems disciplines to promote these technologies in a cost conscious and value added manner. PMID- 9735452 TI - Clinician usage patterns of a desktop radiology information display application. AB - We developed a system for delivering radiologic images and reports to desktop computers used for the electronic medical record (EMR). This system was used by both primary care physicians and specialists primarily in the out-patient setting. The system records all physician interactions with the application to a database. This usage information was then studied in order to understand the value and requirements of an application that could display radiology information (reports and images) on EMR workstations. In this report we describe some of the differences and similarities in usage patterns for the two physician groups. A very high percentage of physicians indicated that having image display capabilities on the workstations was very valuable. PMID- 9735454 TI - Comparison of three display methods for evaluating CT angiography data for the vascular assessment of renal donors. PMID- 9735455 TI - Impact of filmless imaging on the frequency of clinician review of radiology images. AB - The purpose of this study was to determine the impact of filmless imaging on the frequency with which physicians access radiology images and to assess clinician perception of image accessibility using a hospital-wide Picture Archival and Communication System (PACS). Quantitative data were collected at the Baltimore VA Medical Center (BVAMC), prior to and after conversion to filmless imaging, to determine the frequency with which clinicians access radiology images. Survey data were also collected to assess physician preferences of image accessibility, time management, and overall patient care when comparing filmless and film-based modes of operation. In general, there was a significant increase in the average number of radiology images reviewed by clinicians throughout the hospital. However, the one are in the hospital where this trend was not observed was in the intensive care unit (ICU), where the frequency of image assess was similar between film and filmless operations. Ninety-eight percent of clinicians surveyed reported improved accessibility of images in a filmless environment resulting in improved time management. The mean clinician estimate of time saved due to the use of PACS was 44 minutes. The study documented a combination of clinician perception of improved accessibility and substantial time savings with the use of a hospital-wide PACS, which was supported by objective measurements. The increased frequency of image review by clinicians and rapid image access should provide a further impetus to radiologists to decrease report turnaround time to provided "added value" for patient care. PMID- 9735456 TI - Evaluation of commercial PC-based DICOM image viewer. PMID- 9735457 TI - Challenges for pediatric radiology using computed radiography. PMID- 9735458 TI - PACS and CR implementation in a level I trauma center emergency department. AB - Implementation of a picture archive and communication system (PACS) at a large teaching hospital is an expensive and daunting endeavor. The approach taken at the University of Alabama Hospitals has been to assemble an institution-wide system through focused integration of smaller mini-PACS. Recently a mini-PACS using Computed Radiography (CR) has been placed in the Emergency Department (ED) of a Level I Trauma Center completely replacing conventional screen-film radiography. This area of the hospital produces approximately 250 images per day and provided many challenging requirements: the need for rapid radiography; providing good image quality for difficult examinations with potentially uncooperative patients; reproduction of lost films to maintain availability of images to multiple consulting teams; and frequently unknown patient demographics. The PACS includes both vendor-supplied and in-house developed devices for image storage, distribution, and display. Digital images are produced using two photo stimulable phosphor CR systems. Currently, all radiographic examinations are acquired digitally with production of a hard copy film as well as electronic distribution via the PACS. Interpretation of images is done primarily via hard copy with a goal of transition to soft copy interpretation. This paper discusses the functional requirements of the PACS and solutions to workflow issues arising in the ED. PMID- 9735459 TI - Advanced digital mammography. AB - Mammography is the most effective method for early detection of breast cancer and yet, 10% to 30% of women who have breast cancer and undergo mammography have negative mammograms. Furthermore, of the women who are sent to biopsy, only 20% to 40% actually have breast cancer. Quantitative analysis of the radiographic features of microcalcifications and masses may help radiologists improve their specificity. PMID- 9735460 TI - Panoramic dental radiography using a charge-coupled device receptor. AB - Panoramic radiography using a slit beam and film/screen receptor is standard for the emergency room evaluation of mandibular fractures and also in dentistry. This study compared the spatial resolution, area distortion factors, and the dosage considerations for a panoramic system where standard film/screen and a charge coupled device were alternatively employed as the image receptor. Resolution and image contours were determined using a lead resolution grid positioned at selected beam projection angulations. Exposure measurements were carried out using a RANDO average man phantom and a 3 cc beryllium-windowed ionization chamber. The maximum spatial resolution with film approached 5 lp mm-1 whereas with the CCD the maximum resolution was just above 4 lp mm-1. Consequently, the image layer was reduced slightly in width when using the CCD receptor. The use of the CCD resulted in skin exposure reduction exceeding 70%. PMID- 9735461 TI - Clinical trial of panoramic dental radiography using a CCD receptor. AB - The objective of this study was to evaluate the perceived clinical efficacy of a charge-coupled device (CCD) detector for panoramic radiography by comparing the images produced to conventional film/screen radiographs using the same machine and patient population. For clinical evaluation, 18 criteria were selected. These included overall assessment of the area of coverage, clarity of dental structures, clarity of bony outlines, specific anatomic details such as the maxillary sinus floor, mandibular canal and mandibular condyle, and region-by region assessment of the dentition. Observers acted independently using identical optimal viewing conditions. Film and digital radiographs were evaluated separately. A five interval Likert rating scale was used. Digital images were rated superior to the conventional film radiographs for 14 criteria. Film radiographs marginally outperformed digital images for three criteria. For one criterion (periodontal bone status) the two modalities showed no difference in terms of the means ratings. It was concluded that digital images are clinically equivalent to conventional film/screen images for panoramic dental radiography. PMID- 9735462 TI - Direct digital versus conventional film screen radiography of the musculoskeletal system. PMID- 9735463 TI - Copper doped alkali halides for computed radiography and digital imaging. AB - We report here the results of our x-ray fluorescence, photostimulated luminescence, and time resolved laser spectroscopy studies in KCI:Cu. This material seems to possess some desirable properties for being used as an imaging plate in computed radiography. This is a US government work. There are no restrictions on its use. PMID- 9735464 TI - Quantitative evaluation of overall electronic display quality. PMID- 9735465 TI - Image Quality of CRT displays and the effect of brightness of diagnosis of mammograms. PMID- 9735466 TI - Improved sensitivity and specificity of mammograms by producing uniform luminance from viewboxes. PMID- 9735467 TI - The digital film viewer: a novel technology for optimizing film-based radiology. PMID- 9735468 TI - Early clinical experience on cone-beam CT. PMID- 9735469 TI - Getting the picture: when the wide-band telecommunications infrastructure will be widely available. PMID- 9735471 TI - 9th International Genome Sequencing and Analysis Conference. Hilton Head, South Carolina, USA. September 13-16, 1997. Abstracts. PMID- 9735470 TI - Managing security vulnerabilities in a networked world. PMID- 9735472 TI - Quality management for the Medicare generation. PMID- 9735473 TI - A comparison of expectations for selected physician and managed health care services between Medicare-eligible and the next Medicare-eligible generation. AB - The year 1998 marks the beginning of a major turning point in the composition of the Medicare-eligible population, making the next few years a significant watershed for those concerned with the provision of health care to seniors. For approximately the next 12 years, those seniors entering the Medicare-eligible health care market, the so-called Swing generation, will have different characteristics and expectations from those generations that precede or follow it. Through standard survey methods, this study explores and compares the differences in selected health services expectations between the current generation of Medicare-eligible seniors and the next Medicare-eligible generation. Significant differences were found between the two groups on variables related to wait time, medical preparedness, communication, and expectations of managed care coverage. The authors discuss the implications of the findings for health care service providers and managed care organizations. PMID- 9735474 TI - The ethical challenge and the futile treatment in the older population admitted to the intensive care unit. AB - Projections for the future suggest that the United States population will grow by 10-15% by the year 2000, but the number of people over the age of 80 will increase by 66%. As a result, the increase that has already been observed in the number of elderly patients requiring major medical attention can only be expected to grow. This study reviews the admissions to the intensive care unit (ICU) over the last 5 years by age to analyze whether the ICU admissions are higher for the patients older than 60 years of age. We considered all the admissions to the surgical (SICU) and medical (MICU) intensive care units at Morristown Memorial Hospital from January 1, 1992, to December 31, 1996. Patients were divided into age brackets (0-9 years, 10-19 years, 20-29 years,... > 90 years) and by gender. Medical and surgical admissions were analyzed including the average length of stay in the ICU. Daily charge for bed occupancy was reviewed based on the hospital data reported in 1995. The death rate was also considered. Fisher corrected chi 2 and a Student t test were used for statistical analysis. A total of 6243 patients (2926 female and 3317 male) were admitted to the ICU over the 5 year period. The ICU admissions rate was higher in patients above 60 years of age compared with those below 60 (60% versus 30%, respectively). The age group with the highest admissions rate was between 70 and 79 years, followed by the 60-69 year group. These two groups had significantly more admission than all other groups (P < 0.001). Medical patients' length of stay was shorter than the surgical group, and they had a lower rate of admission to the ICU. The death rate was higher in the group older than 60 years. They also spent a longer time in ICU compared with the younger group (22 +/- 7 days versus 12 +/- 8 days). The charge per day per bed was $2100 in the ICU, $1600 in a telemetry floor, and $950 in a regular floor. The charge per bed in the group above 60 years old was double compared with the one for the younger group. Older patients were admitted to the ICU with a significantly higher frequency than was the younger group. There were more surgical than medical patients admitted to the ICU. The mortality rate and the daily cost, based on daily bed charge, was significantly higher in the older group. Based on our experience, older people had a more difficult recovery in ICU than did the younger people. In our opinion we should treat acute critical illness but not terminal pathology. A problem exists in educating physicians about which patients will derive no benefit from the ICU. This will determine if we can decrease or avoid the use of the ICU and its accompanying expense, in situations where it does not significantly increase survival and the quality of life. PMID- 9735476 TI - Redesign for effective quality assurance: pilot program in an urban community hospital. AB - An urban community hospital redesigned the quality assurance (QA) procedures of its medical staff to increase the effectiveness of QA. The best features of the plans of several leading hospitals were presented to departments and committees. These redesigned their QA efforts into a hospital-wide plan. Leadership commitment to redesign, increased process objectivity, and more horizontal structure resulted in the identification of adverse events and the assignment of numerical assessments to them. The rate of adverse event detection was equal to the rate achieved by the literature's benchmark study. PMID- 9735475 TI - Managed care for elderly people: a compendium of findings. AB - Although managed care seems to serve well the interests of non-elderly enrollees and their payers, elderly people face more risks. Chronic conditions, multiple problems, and more limited resources make them more vulnerable, whereas multiple payer sources make them more complicated to cover. This synthesis of managed care delivered in Medicare and Medicaid demonstration projects serving elderly beneficiaries shows that managed care plans either select or attract enrollees who suffer fewer frailties than those served in fee-for-service settings, exhibit reluctance to enter rural markets, provide a broad range of elderly-specific services, offer more comprehensive coverage and services, and result in greater perceived access problems, particularly for vulnerable subgroups. Plans operate more cheaply by using fewer resources, even after adjusting for case mix differences. Managed care enrollees tend to be more satisfied with financial and coverage aspects, whereas fee-for-service enrollees report higher satisfaction on other dimensions. In acute care settings, process of care findings were mixed, whereas clinical and self-reported outcome indicators were no better and in some instances worse in managed care. Long-term care enrollees, in the few studies reported, consistently faired worse in both the processes and outcomes of care. These findings suggest that further research on the effects of managed care in its rapidly changing incarnations is needed, particularly with respect to how to improve the quality of acute and long-term care delivered to elderly people and the proper role of government and other key actors in the health care system. PMID- 9735477 TI - Comparing the value of service between a state hospital and a private, for-profit psychiatric hospital: a clarified role for tertiary care. AB - We apply pattern-recognizing artificial neural networks (ANNs) to the patients of two psychiatric hospitals, a private, for-profit hospital (PH) and a state hospital (SH), both serving the southern tier of Maine (approximately two-thirds of the state's population, i.e., 800,000 persons) over a 19-month period. In our data from the PH, N = 837 admissions, and at the SH, N = 834. Unique patient identifiers were assigned to patients so that their individual patterns of care could be incorporated into our ANNs. We used a previously reported methodology to measure quality of care (Q), and developed a measure of value of service (V) from the patients' perspective for both facilities. A random portion of the demographic and outcome data of patients from each hospital was sequestered as a test set, whereas the remainder was used to train ANNs with length-of-stay (LOS) as an outcome measure. Q, and V normalized for risk (RR), i.e., V/RR, were calculated for each test set, which included multiple admissions of individual patients to each hospital. The methodology for V accounts for the severity of illness with the calculation of a metric called U/G, for the differences in case mix by exchanging "virtual patients" between ANNs, and for entropy in the health care system by using a metric called the risk ratio (RR). Results showed that V/RR was 2.4 times greater at the SH than at the PH. This advantage is likely due to prior knowledge of individual patient patterns of treatment by the SH's staff. Data sets from each hospital using only single admissions for each patient in the study period (thereby eliminating unique patient patterns of LOS), yielded a V/RR that was only 21% greater at the SH. We hypothesize that this difference is due to the SH's ability to use treatment and discharge based upon patient strengths and level of clinical improvement, unfettered by insurance deadlines. Approximately 5% of the admissions to our studied PH went on to our SH, namely, the most severely impaired and indigent patients. The SH not only had twice the value, but also had half the cost of the PH, despite the greater number of treatment non-responders and non-compliants at the SH. The reasons for this are skilled staff and specialized ancillary services, staff knowledge of patients' responses to previous therapy, and individualized LOSs. These are features that create a caring environment and better compliance with treatment. This study emphasizes the value of tertiary care psychiatric facilities in a comprehensive mental health care delivery system. There are few studies of the effects of excessive downsizing of SHs on the community, but reports from Massachusetts (in 1995) suggest a 79% increase in suicides when the closing of SH beds exceeded 98% of maximum historical census. Routine use of pattern-recognizing tools such as ANNs would serve to inform the public about the value of mental health services so that the most vulnerable in our society are not neglected. PMID- 9735478 TI - Continuous quality improvement, total quality management, and reengineering: one hospital's continuous quality improvement journey. AB - In recent years, there has been significantly increasing interest in the application of continuous quality improvement (CQI) and total quality management (TQM) in the health care arena. This case analysis is designed to identify and assess the strategies and processes that led to the successful implementation of CQI in the Emergency Care Center at St. Mary's Hospital in Grand Rapids, MI. PMID- 9735479 TI - Commentary: patient education presentations. AB - Medical students often learn how to teach through observation of residents and attendings. The project described enables them to actively teach groups of patients, and allows them to begin developing their own style of teaching. It also demonstrates to the students that teaching is a skill to be learned, and methods may vary tremendously. PMID- 9735480 TI - A case of cultural misunderstanding. PMID- 9735481 TI - Managing sore throats. PMID- 9735482 TI - Thrush during lactation. PMID- 9735483 TI - Management of the paediatric foreskin. PMID- 9735484 TI - Is it allergy? AB - BACKGROUND: 'Allergy' refers to disorders involving IgE antibody. The cross linking of IgE molecules on mast cells by an allergen releases mediators that produce the typical allergic reaction. OBJECTIVE: To distinguish allergies from other disorders, particularly those where mast cells are also involved. DISCUSSION: Many non allergic processes can mimic allergy. A careful history and examination and discriminate use of investigations is important to make the distinction. PMID- 9735485 TI - Allergy in the airway. AB - BACKGROUND: Airway disease is responsible for significant morbidity worldwide and the role of allergies in the cause and persistence of airway symptoms is becoming increasingly appreciated. OBJECTIVE: The exposure of the airway to allergens depends on the size of inhaled particles. Common allergens encountered in Australia include grass pollens, house dust mites and animal danders and sensitivity to each leads to a different clinical pattern of disease. The diagnosis of allergy requires a history of symptoms related to exposure to an allergen, together with detection of allergen-specific IgE. The evidence for the role of allergens in contributing to allergic airway disease is extensive. DISCUSSION: Airway allergies are an important factor in the genesis of asthma and allergic rhinitis. There is increasing evidence that modification of exposure to allergens can improve allergic symptoms and may prevent allergic diseases. Methods of allergen avoidance are discussed. PMID- 9735486 TI - Successful immunotherapy. A partnership between consultant and general practitioner. AB - BACKGROUND: Allergen-specific immunotherapy has been used to treat allergic patients for many decades. Numerous clinical trials have demonstrated its efficacy in patients with seasonal allergic rhinitis, perennial allergic rhinitis, stinging insect allergy and allergic asthma. OBJECTIVE: This article discusses antigen specific immunotherapy and outlines the conditions that respond well to it. DISCUSSION: Immunotherapy has the potential to cause immediate systemic reactions which rarely may result in fatal anaphylaxis. Therefore, only practitioners with appropriate training and expertise should make the decision to prescribe immunotherapy based on careful patient selection and extract prescription. Patients receiving this therapy require supervision in a setting with the necessary resuscitation equipment by a practitioner with the skills to recognise and treat adverse events. PMID- 9735488 TI - Making sense of the results of your research. AB - The key to making the most of your results lies in clarifying your plan of analysis first. Cleaning your data is also essential. Make sure you have help and that you give credit to any funding sources. PMID- 9735489 TI - The antiphospholipid antibody syndrome. AB - BACKGROUND: Recurrent miscarriages cause enormous distress and despair for sufferers. In a small number of patients this condition forms part of the antiphospholipid antibody syndrome. The anticardiolipin antibodies became quantifiable in 1983, which has meant that those suffering with this condition could be identified. OBJECTIVE: This article reviews the current understanding of this condition and offers an approach for managing patients with various components of this syndrome. DISCUSSION: The decision to screen for these antibodies is sometimes difficult as they can be presented in the normal population. However, by identifying the patient suffering with this condition, treatment can offered which may prevent a further miscarriage or a pregnant patient suffering a thromboembolic episode. PMID- 9735490 TI - Alpha adrenergic blockers and adrenaline. A mysterious collapse. AB - In 1995 I encountered a problem with the use of adrenaline in the resuscitation of a patient on prazosin who had an anaphylactic collapse resulting from a bee sting. I concluded that the problem was the result of the 'reversed adrenaline effect' and I predicted that other drugs with alpha adrenergic receptor blocking activity such as the phenothiazines could similarly obstruct the action of adrenaline. A literature review of the reversed adrenaline effect revealed a number of animals studies but no articles relating to humans. I present this case study as another example of drugs with alpha adrenergic receptor blocking activity (thioridazine and amitriptyline) obstructing the action of adrenaline in anaphylactic collapse. PMID- 9735491 TI - Challenges facing primary care mental health in Australia. AB - BACKGROUND: Changes to the organisation of mental health services in Australia and a recognition that general practitioners deliver the great bulk of mental health services to the community have necessitated a re-examination of the role of general practitioners in the clinical area. OBJECTIVE: This article examines controversies in mental health care such as the difficulties with detection, the nature of general practice psychiatry, the inappropriateness of undergraduate training, workforce related issues, mental health services in rural areas and continuing medical education. DISCUSSION: General practice based services for people with mental disorders are distinct in many respects from specialist psychiatric services. Systemic changes need to be made to training for general practitioners, to incentives for delivery of complex care and to the way in which the different health care sectors work together. PMID- 9735492 TI - Headache and drowsiness. PMID- 9735493 TI - Golfer's heel. PMID- 9735494 TI - The misplaced left hook. PMID- 9735495 TI - Patient education. House dust mite allergy. PMID- 9735496 TI - Paediatric dermatology. Impetigo. PMID- 9735497 TI - Practice tip. Ingrown toenails. PMID- 9735498 TI - [The cochlear implant. Study in the profoundly deaf adult from the standpoint of quality of life and therapeutic cost (QALY Index of Quality)]. AB - Cost utility analysis is a method of cost-effectiveness analysis which provides results in terms of cost per quality-adjusted life-year (QALY). This method is progressively largely used for medical technology assessment. It permits cost effectiveness comparisons between medical interventions. The cost per QALY of cochlear implant was determined in 30 adult patients suffering from acquired profound deafness. This study indicates that this technology provides significant improvements and is quite cost-effective. However, as far as profound deafness is concerned, the reliability of QALY needs to be improved. PMID- 9735499 TI - [Cutaneous lymphoma of the Sezary and Bouvrain type. Progress in physiopathologic and therapeutic outlook]. AB - In 1938, Sezary and Bouvrain reported a new form of disseminated cutaneous lymphomas, characterized by an erythrodermia, and by the presence in the blood of cells with a cerebriform nucleus. It was shown later that this disease was related to the proliferation of a subpopulation of CD4+ T lymphocytes. With the aim to develop new therapeutic strategies for these lymphomas, we tried to evidence and to characterize the tumor-specific immune responses in these cutaneous T-cell lymphomas. We have isolated several T cell clones from lymphocytes infiltrating a human MHC class II negative cutaneous T cell lymphoma. We describe here two of these clones, TC5 and TC7, with respectively a CD4+CD8+ and CD4+CD8- phenotype. Both clones mediated a specific MHC class I-restricted cytotoxic activity toward the fresh autologous tumor cells, and autologous tumor cell lines previously established with IL-2 and IL-7 from the skin and from the blood. Analysis of the T-cell receptor V beta gene expression revealed that the tumor cells, that were shown to have a trisomy 7 by fluorescent in situ hybridization, expressed V beta 7/J beta 2.3, V beta 13/J beta 2.5 and V beta 22/J beta 2.5 rearrangements. Phenotypic analysis using specific anti-V beta monoclonal antibodies indicated that only V beta 13 could be detected on the cell membrane of the tumor cells. Analysis of the TCR V beta gene expression of the clones showed that TC5 and TC7 expressed a unique TCR-V beta transcript, corresponding respectively to V beta 5/J beta 2.3 and V beta 17/J beta 2.7 gene segments. To determine whether these reactive T lymphocytes were present in vivo, we used specific primers corresponding to TC5 and TC7-V beta TCR transcripts. The results demonstrated that both cytotoxic T cell clones were present at the lesional skin site and amplified in vitro. TC7 was found in the patient peripheral blood invaded by tumoral cells, whereas TC5 was not, indicating that the repertoire of the reactional lymphocytes differs in the blood and at the tumor site. These results demonstrate for the first time the presence of reactive T lymphocytes with CD4 or double positive phenotype infiltrating a CTCL. These findings raise new perspectives for the treatment of cutaneous T-cell lymphomas. PMID- 9735500 TI - [The requisite internationalization of the campaign against smoking]. AB - One smoker out of two dies from smoking. Fifty five percent of the French population smoke at age 18. To decrease the tobacco consumption is thus a paramount public health objective. Following WHO and "Europe Against Cancer", a comprehensive program against smoking should ban advertising, increase prices, protect non-smokers, educate children, promote non-smoking behaviors and help smokers who want to stop. The measures taken in France, following the law of January 10th, 1991, made possible a 11.1% decrease of tobacco consumption and a 14.5% decrease of cigarette consumption. If France is exemplary for the ban of tobacco advertising, the means devoted to education and information are poor, the weakest in Europe. So, it is still possible to speed up the decrease of tobacco consumption in France so that the estimate of 165,000 deaths related to tobacco consumption in 2025 will not occur. This success has to be interpreted within a European and Worldwide context. The tobacco industry is responsible for denying the danger of tobacco, the addictive effect and nicotine and for targeting children and adolescents. Faced to the forecast of 100 million deaths within twenty years, only an international solution can be of the right size. WHO should be given this responsibility. PMID- 9735501 TI - [Management of 29 children with thyroid cancer following the Chernobyl accident]. AB - As a consequence of the Chernobyl nuclear power plant accident, a considerable increase of thyroid cancer among contaminated children has been reported in Ukraine, Belarus and Russia. A group of 29 children aged from 5 months to 10 years (mean 4.7 years) at the time of the accident, with a papillary thyroid cancer, have been examined at the Pitie-Salpetriere hospital in Paris. The cancer was discovered by systematic ultrasonography in only 25% of cases. No reliable dosimetric estimation was achieved. The initial surgical treatment was performed in Ukraine. Cervical lymph node and pulmonary metastases were present in 24 and 11 cases respectively. A complementary surgical treatment was necessary for 9 children and one to four radioiodine treatments were given to 24 children. With a mean delay of 7 years after the cancer discovery, an apparent cure or a remission was obtained for 20 children, 6 children have cervical lymph node metastases requiring a surgical treatment and 3 have evolving lung metastases. The management of the great number of foreseeable cases of thyroid cancer requires an improved systematic screening, a large number of rooms dedicated to high activity radioiodine treatments, funds for disposable material and training missions. PMID- 9735502 TI - [Factors predictive of early virologic response after a first treatment with an protease inhibitor in the course of HIV infection]. AB - As many factors may be involved in therapeutic response to triple therapy with protease inhibitor (PI), the aim of this study was to determine the influence of epidemiological factors (sex, risk factors), clinical status (previous number of AIDS defining events), immunological status (baseline CD4 T cells count), virological factor (baseline viral load), previous antiretroviral therapy and duration of AZT therapy (> or < 6 months), number of prescribed reverse transcriptase inhibitors (RTI), therapeutic strategy (switch to different RTI or only addition of PI) and compliance, on early virological response (M2-M3) after initiation of triple therapy with PI. These results concerned 167 patients treated with triple therapy including PI. A viral load response was defined in three types: complete response (undetectable: < 500 copies/ml) for 100 patients; partial response (significant decrease: > 0.5 log from baseline) for 30 patients and no response for 37 patients. Only two parameters were associated of good virological response: no previous antiretroviral therapy (p < 0.001) and good compliance (p < 0.001). No significant difference was observed between patients with no prior therapy and pretreated patients, in terms of median baseline CD4 count and observance. The baseline median viral load was higher in naive patients despite a better response. In pretreated patients, the type of response appeared to be dependent on the duration of AZT treatment (p = 0.06) and good compliance (p = 0.06). Among the 100 patients with initial complete response, only 23/81 were still undetectable after a median of 13 months of therapy. PMID- 9735503 TI - [Distribution of bone metastases of cancers. A scintigraphic study of 376 cases]. AB - Technetium 99m methylene bisphosphonate bone scans of 376 patients with cancers of breast, prostate, lung, kidney, colon, and bladder and ENT cancer were reviewed, and the distribution of skeletal metastases was analyzed. Differences were not significant for rank order of metastatic involvement in 9 selected regions in any cancer, but the breast carcinoma. Patients with breast cancer had less pelvis and more skull involvement. The rate of skull metastases was significantly higher in breast cancer than in prostatic (p < 0.001), lung (p < 0.01) and kidney (p < 0.05) cancers. These results are weakly demonstrative of a role of the vertebral veins in hematogenous spread of breast and prostate cancer. The hypothesis is proposed that this pattern of dissemination, which is suggested by experimental and clinical features, is poorly perceptible in the distribution pattern of skeletal metastases because of the concurrent arterial spread of breast and prostatic tumors cells, which is likely preponderant. PMID- 9735504 TI - [Functions of neutrophil motility of human blood: locomotion and chemotaxis in simplified systems]. AB - Polymorphonuclear leukocytes (PMN, granulocytes) employ their plasma membranes and subjacent microfilament-rich peripheral cytoplasm for such motile functions as adherence and spreading, random locomotion, chemotaxis (directed locomotion), and phagocytosis. All of these functions are preserved in certain anucleate, granule-poor, cytoplasmic fragments (cytoplasts) derived from PMN. Thus, the sensing, transduncing, and effector capacities involved in these functions remain integrated without control from nuclei or from the other cellular organelles left behind when the cytoplast forms. More recently, we have begun to examine in intact PMN the role of divalent cations, which have been thought to be essential for motile function of leukocytes in general, and for the function of critical adhesion molecules in particular. In slide preparations under direct microscopic observation, EDTA (10 mM; to chelate divalent cations) did not impair either random locomotion or chemotaxis, nor did specific antibodies to beta-2 (CD 18) integrins or to other PMN integrins. Motile behavior appeared to benefit from the close approximation of slide and coverslip ("chimneying"). Thus, in "close quarters", PMN can generate the force for locomotion even when adhesion molecules are lacking or disabled. We relate these findings to the reported independence from integrins of PMN in certain experimental and diseases states. PMID- 9735505 TI - [The movement of the human spermatozoon]. AB - The human sperm flagellum is composed by an axoneme made up of peripheral doublets of longitudinal microtubules on which are fixed dynein arms and radial spokes. A mechanochemical cycle of attachment-detachment of dynein arms on the adjacent microtubules results in a sliding/bending or microtubules. This process repeated along the flagellum, induces the propagation of a wave. The progressive movement of ejaculated spermatozoa is modified during their transport through the feminal genital tract under the influence of microenvironmental factors. The microfibrillar structure of cervical mucus constraints the flagellar wave amplitude whereas the tubal or follicular secretions induce an hyperactivation of the movement characterized by a loss of progression and a high degree of flagellar curvature. Sperm movement modifications are depending on external regulatory factors among which some could be secreted by the oocyte or by the perioocyte layers to create a sperm chemotaxis which would optimize the gametic interaction. The external factors modulate sperm movement through their interaction with membrane "receptors" and intracellular messengers such as cyclic AMP, ATP, Calcium, pH. These latter control dynein-microtubules interaction through a phosphorylation-dephosphorylation process of axonemal proteins. PMID- 9735506 TI - [Telemedicine and telepathology in hematology]. AB - Long distance transmission by the telephone network have offered new facilities in transmitting images in real time. Pathologists and other specialists like radiologists are now using these transmissions of digitalised pictures, both for diagnostic support and for constituting image data base for teaching purposes. Haematological application of this new method is shown as a particularly good example. PMID- 9735507 TI - Use of methotrexate in children. AB - Methotrexate continues to be the safest and most efficacious second-line drug for the treatment of JRA. In addition, it is useful in other inflammatory conditions in children. Careful education is necessary, particularly with regard to the importance of laboratory tests and the avoidance of comorbidity such as pregnancy and alcohol-induced liver injury. Health care providers should be comfortable discussing these issues with children and adolescents. PMID- 9735508 TI - The eye and rheumatic disease. PMID- 9735509 TI - An Advisory Committee Statement (ACS). Committee to Advise on Tropical Medicine and Travel (CATMAT). Statement on Japanese encephalitis vaccine. PMID- 9735510 TI - CPHA partnership renewed in the new national HIV/AIDS strategy. PMID- 9735511 TI - Protecting our children. PMID- 9735512 TI - The Ontario ban on smoking on school property: perceived impact on smoking. AB - We evaluated the impact of the November 1994 ban on smoking on school property in Ontario. Telephone interviews were conducted at the end of the 1995-96 school year with 213 high school administrators. Almost all high schools (96%) prohibit smoking on school property. Although some smoking still occurs on school property, the location of smoking by students has changed, giving rise to perceptions of both benefits and risks, as well as varying complaints from parents, students, neighbours, and nearby businesses. Most of the problems arising from the ban are viewed as minor. Local conditions, particularly the geographic environment of the school, appear to be important determinants of complaints and problems. While sizeable minorities of school administrators felt the ban had favourable effects, the majority perceived little effect on either smoking behaviour or attitudes towards smoking. PMID- 9735513 TI - Urban air pollution and mortality. PMID- 9735514 TI - The Ontario ban on smoking on school property: issues and challenges in enforcement. AB - We document implementation and enforcement activities undertaken by high schools and health units with regard to the 1994 ban on smoking on school property in Ontario. Telephone interviews were conducted in the early summer of 1996 with 213 high school administrators and 38 tobacco enforcement personnel in health units. While some schools are unclear about enforcement responsibility, most are making efforts to enforce the ban, including warning and suspending students. Some school administrators (30%) suggest the reinstitution of designated smoking areas on school property. One quarter of health units had not made enforcement visits in schools in the 1995-96 school year and a minority accounted for most of the warnings and tickets issued to students. While most tobacco enforcement officers perceive that schools support the ban, they report some problems in obtaining cooperation in enforcement. However, only 11% suggest returning to designated smoking areas on school property. PMID- 9735515 TI - The epidemiology of cocaine and opiate abuse in urban Canada. AB - This study describes the epidemiology of cocaine and heroin abuse in urban Canada as part of an initial report on a national substance abuse surveillance system, the Canadian Community Epidemiology Network on Drug Use. Data pertaining to prevalence of use, law enforcement, treatment, morbidity and mortality of cocaine and heroin were obtained from the appropriate health and law enforcement institutions in six sentinel cities: Vancouver, Calgary, Winnipeg, Toronto, Montreal and Halifax. Cocaine and heroin appear to be more available in Vancouver than in the remaining cities. In all CCENDU cities, large proportions of persons in treatment programs for substance abuse identified cocaine as their major addiction; however, there is considerable variation in treatment utilization regarding heroin. Vancouver ranks first in terms of the per capita number of cocaine- and heroin-related hospital separations and mortality rate. Cocaine abuse appears to be an emerging problem in Calgary, Winnipeg and Halifax, and opiate abuse appears to be an emerging problem in Calgary. PMID- 9735516 TI - Injection drug use among street youth: a dynamic process. PMID- 9735517 TI - Review of Canadian low-risk drinking guidelines and their effectiveness. AB - This study compared 18 low-risk drinking guidelines that were gathered from Canadian government agencies, non-government agencies, medical bodies, and public and private agencies involved in the treatment of addictions. The results show that two sets of guidelines are predominantly used in Ontario. The formulation of these guidelines was entirely independent and their intended audiences are also different. However, a direct comparison of the two guidelines shows that differences are more apparent than real. This study also examines the literature evaluating low-risk drinking guidelines Very little literature exists on evaluating low-risk drinking guidelines as vehicles for primary prevention and it is not known to what extent such guidelines influence knowledge and drinking behaviour. Future low-risk drinking recommendations should be evaluated for knowledge about standard drink units, awareness of the guidelines, use of materials and aids included in the dissemination program, and changes in behaviour from campaign exposure. PMID- 9735518 TI - Syphilis in an urban community. AB - Recent reports of changes in the epidemiology of syphilis prompted a review of syphilis in our urban community. All records of positive syphilis serology reported to the City of Scarborough Health Department between 1990-94 were reviewed for key epidemiological variables. While infectious stages of syphilis were reported more often among young adults, incidence for all stages increased among successive age groups, with a male/female ratio of 1.0. One in five cases were identified during immigration screening, with a disproportionate number of cases immigrating from the Caribbean, Africa and Subcontinental India. Overall, the incidence of syphilis decreased during the study. However, a correlation of 0.95 was found between the provincial incidence of syphilis and number of tests ordered. The observed decrease in syphilis, therefore, may represent a decrease in detection owing to lack of testing. PMID- 9735519 TI - A descriptive study of verocytotoxigenic Escherichia coli (VTEC) cases reported in Ontario, 1990-1994. AB - Characteristics of VTEC cases identified through routine surveillance in Ontario between 1990 and 1994 are described. Information was extracted from the Reportable Disease Information System (RDIS) of Ontario and was evaluated for its completeness and internal validity. A total of 2,441 VTEC cases were identified for the five-year study period corresponding to an average annual rate of 4.8 cases per 100,000. Sixteen deaths were recorded. Bloody diarrhea was reported for 546 patients (40%) and was the most frequently reported symptom. For most cases, the home was recorded as the likely risk setting (36%). Food was incriminated as the source of infection for more than 36% of cases. Nine (69%) of the thirteen data fields compulsory for transmission to the Ontario Ministry of Health had less than 10% of combined missing and unspecified values. Fields describing risk factors had greater than 56% of entries missing or unspecified. PMID- 9735520 TI - Value of ecological analysis. PMID- 9735521 TI - North American liver fluke (Metorchis conjunctus) in a Canadian aboriginal population: a submerging human pathogen? PMID- 9735522 TI - Education of women about the prevention of preterm birth. AB - OBJECTIVES: To describe: 1) The education of pregnant women by health care professionals about the prevention of preterm birth; and 2) professionals' views about future initiatives. BACKGROUND: A population survey of health professionals was conducted in Eastern Ontario. The response rate was 73% (608/835). RESULTS: Education materials for women receiving prenatal care about the prevention of preterm birth were available from 10% (12/115) of family physicians, 40% (23/58) of obstetricians, 19% (57/306) of labour room nurses and 76% (94/124) of the prenatal teachers. Only one third of physicians routinely discussed the signs and symptoms of preterm labour prior to 20 weeks. Practitioners' future priorities were smoking cessation programs for pregnant women and increased attendance at early prenatal classes. CONCLUSIONS: Most women are not being educated by anyone in the health care team about the prevention of preterm birth. There is a need for multidisciplinary guidelines about the timing and type of information for women about risk reduction and the early identification and treatment of preterm labour. PMID- 9735523 TI - [Smoking cessation for hospitalized patients: a quasi-experimental study in Quebec]. AB - PURPOSE: The purpose of this study was to evaluate the impact of a smoking cessation intervention for hospitalized patients, implemented by regular staff and incorporated into their routine care of patients. METHODS: The intervention was conducted in one experimental hospital and in two control hospitals. RESULTS: One year after discharge, 15% of smokers became non-smokers in the experimental hospital versus 8% in the control hospitals. This difference is not statistically significant (p = 0.08), however a small sample in the control hospitals had an influence on the statistical power. A logistic regression highlights program participation as the only variable predictive of a non-smoker status one year after discharge, considering both types of hospitals (experimental and control). CONCLUSIONS: Establishing relevancy of smoking cessation intervention for hospitalized patients is probably no longer needed. But research should be carried on towards finding better ways to convince the staff to intervene, towards establishing relevancy for specialized staff and defining intensity of required interventions before and after hospitalization. PMID- 9735524 TI - Creating a population-based linked health database: a new resource for health services research. AB - As the availability of both health utilization and outcome information becomes increasingly important to health care researchers and policy makers, the ability to link person-specific health data becomes a critical objective. The integration of population-based administrative health databases has been realized in British Columbia by constructing an historical file of all persons registered with the health care system, and by probabilistically linking various program files to this 'coordinating' file. The linkages have achieved a high rate of success in matching service events to person-specific registration records. This success has allowed research projects to be proposed which would otherwise not have been feasible, and has initiated the development of policies and procedures regarding research access to linked data. These policies and procedures include a framework for addressing the ethical issues surrounding data linkage. With continued attention to confidentiality issues, these linked data present a valuable resource for health services research and planning. PMID- 9735525 TI - [Socioeconomic variables and the prevalence of dental caries in second and sixth grade Quebec children in 1989-90]. AB - OBJECTIVES: 1) To determine caries risk factors in second and sixth grade Quebec children; 2)To test multivariate models which identify children as belonging to a high prevalence group. METHODS: For the 1989-90 Sante Dentaire Quebec survey, 2,291 second grade and 2,111 sixth grade school children responded to a questionnaire on their personal habits of hygiene and diet and underwent a clinical examination, while their parents answered a questionnaire regarding their family's socio-economic status. RESULTS: Statistics demonstrate a stronger link between socioeconomic variables and caries prevalence than demographic and sanitary factors. Children emerging from a high socioeconomic milieu have better dental health than children with low socioeconomic standing. The most effective model, However, registers a sensitivity of 65% and a specificity of 66%, revealing the inadequacy of statistical models to accurately identify children in the caries high prevalence group. PMID- 9735526 TI - Elective neurosurgical office consultations: how well are patients informed? AB - A prospective study was performed on elective neurosurgical office referrals to one neurosurgeon at the Toronto Hospital from September 1988 to May 1996. Patient level of information was tested using the chisquare test on the study population of 2,017 patients grouped according to the type of referring doctor and regional category of diagnosis. There was a statistically significant difference in the degree of knowledge of referred patients according to the type of referring doctor (p < 0.008). In addition there was statistical significance found in the difference in degree of knowledge among patients referred with the various regional categories of disease (p < 0.008). Patients referred by family physicians are not as informed as to the nature of their neurosurgical referral as those referred by neurologists and other specialists. Furthermore, patients with an intracranial diagnosis had a greater level of knowledge about their referral than those patients referred for spinal, peripheral nerve or other diagnoses. PMID- 9735527 TI - Developmental follow-up of 6-7 year old children of mothers employed during their infancies. AB - This study followed up 92 children at ages 6-7 first studied as one-year-olds in order to determine differences in the developmental outcome of the offspring of employed as compared to "stay at home" mothers. The developmental domain evaluated in the 6-7 year old children was peer competence, rated by laboratory play and child psychological test instruments. Mothers' reports of children's behavioral pathology at age 6-7 correlated with a higher number of maternal work hours during the infant's first year, but the children did better during overall play situations. Age 1 attachment ratings better predicted free play social competence for the entire sample than did maternal hours of work absence, but girls accounted for statistical significance. Maternal sensitivity from infancy was associated with maternal reports of (low) problem behaviors; however, regression analyses did not support the hypothesis that relations between either work status or attachment and current problem behaviors were mediated by early maternal sensitivity. PMID- 9735528 TI - Clinical and cultural issues in diagnosing and treating child victims of peer abuse. AB - Examined are clinical and cultural issues in assessing and treating child victims of peer abuse. Profiles for both victims and perpetrators are discussed as are clinical patterns that may emerge in adulthood. Clinical trauma accommodation is discussed to offer the processing and adaptation to peer victimization. A clinical algorithm is discussed to provide the clinician with a pathway for assessment, treatment and follow-up for children who experience peer victimization. PMID- 9735529 TI - Characteristics of toddlers and preschoolers exhibiting severe psychiatric disturbance. AB - The present study reports data collected on 137 toddler and preschool children admitted for acute, psychiatric inpatient hospitalization. Utilizing a comprehensive chart review survey developed by the authors, information was obtained regarding presenting problems, family psychiatric history, family legal problems, child maltreatment in order to examine factors contributing to the need for psychiatric hospitalization. Extremely dangerous behaviors were displayed by these youngsters, placing themselves and others at significant risk of harm. The frequency of occurrence of factors that contributed to these severe adjustment difficulties was alarming. Implications regarding service needs for these young children are discussed. PMID- 9735530 TI - Staff perceptions of sexuality-related problems and behaviors of psychiatrically hospitalized children and adolescents. AB - Clinical staff at two hospitals serving children and adolescents were surveyed regarding their observations and experiences regarding the sexuality-related behaviors and issues of patients. Results of this descriptive study indicate that staff frequently encounter a wide range of such problems, including the effects sexual abuse, sexually aggressive or inappropriate behavior, lack of knowledge about basic hygiene and sexual development, pregnancy and contraception, high risk behaviors, and sexually-transmitted diseases, including AIDS. Similarities and differences of the perceptions by staff of these problems are compared across unit types (children's, adolescents, dually-diagnosed, developmentally-disabled). Implications for staff training, clinical policies, and further research are discussed. PMID- 9735531 TI - Behavioral changes in autistic individuals as a result of wearing ambient transitional prism lenses. AB - A double-blind crossover design was used to assess the efficacy of wearing ambient lenses to reduce the behavioral symptoms of autism. Eighteen autistic individuals, ranging in age from 7 to 18 years, participated in the study. Behavior, attention, and orientation were evaluated at 1 1/2 months, 2 months, 3 months, and 4 months. Compared to the placebo condition, the results showed a decrease in behavior problems at the 1 1/2 and 2 month assessment periods and a slight loss of these benefits at the 3 and 4 month assessment periods. These findings support the prediction that ambient lenses, worn without engaging in visual-motor exercises, have positive effects on autistic individuals. PMID- 9735532 TI - Rejoinder to Mordock's critique of the Fort Bragg Evaluation Project: the sample is generalizable and the outcomes are clear. AB - The Fort Bragg Evaluation Project (FBEP) showed that children in a well implemented and expensive continuum of care had no better clinical outcomes than those experiencing more traditional and fragmented services. In an article published in this journal that was critical of the evaluation, Mordock argued that the FBEP results be viewed with skepticism because of what he perceived to be methodological, design, measurement, and analytic failures of this study. We think it is important to respond to Mordock's critique since it contributes to the great reluctance to seriously consider the study's findings and their implications. PMID- 9735533 TI - You can go anywhere from where you are: a response to the rejoinder. PMID- 9735534 TI - Image-based object recognition in man, monkey and machine. AB - Theories of visual object recognition must solve the problem of recognizing 3D objects given that perceivers only receive 2D patterns of light on their retinae. Recent findings from human psychophysics, neurophysiology and machine vision provide converging evidence for 'image-based' models in which objects are represented as collections of viewpoint-specific local features. This approach is contrasted with 'structural-description' models in which objects are represented as configurations of 3D volumes or parts. We then review recent behavioral results that address the biological plausibility of both approaches, a well as some of their computational advantages and limitations. We conclude that, although the image-based approach holds great promise, it has potential pitfalls that may be best overcome by including structural information. Thus, the most viable model of object recognition may be one that incorporates the most appealing aspects of both image-based and structural description theories. PMID- 9735535 TI - Three-dimensional object recognition based on the combination of views. AB - Visual object recognition is complicated by the fact that the same 3D object can give rise to a large variety of projected images that depend on the viewing conditions, such as viewing direction, distance, and illumination. This paper describes a computational approach that uses combinations of a small number of object views to deal with the effects of viewing direction. The first part of the paper is an overview of the approach based on previous work. It is then shown that, in agreement with psychophysical evidence, the view-combination approach can use views of different class members rather than multiple views of a single object, to obtain class-based generalization. A number of extensions to the basic scheme are considered, including the use of non-linear combinations, using 3D versus 2D information, and the role of coarse classification on the way to precise identification. Finally, psychophysical and biological aspects of the view-combination approach are discussed. Compared with approaches that treat object recognition as a symbolic high-level activity, in the view-combination approach the emphasis is on processes that are simpler and pictorial in nature. PMID- 9735536 TI - Recovery of 3D volume from 2-tone images of novel objects. AB - In 2-tone images (e.g., Dallenbach's cow), only two levels of brightness are used to convey image structure-dark object regions and shadows are turned to black and light regions are light regions are turned white. Despite a lack of shading, hue and texture information, many 2-tone images of familiar objects and scenes are accurately interpreted, even by naive observers. Objects frequently appear fully volumetric and are distinct from their shadows. If perceptual interpretation of 2 tone images is accomplished via bottom-up processes on the basis of geometrical structure projected to the image (e.g., volumetric parts, contour and junction information) novel objects should appear volumetric as readily as their familiar counterparts. We demonstrate that accurate volumetric representations are rarely extracted from 2-tone images of novel objects, even when these objects are constructed from volumetric primitives such as generalized cones (Marr, D., Nishihara, H.K., 1978. Proceedings of the Royal Society London 200, 269-294; Biederman, I. 1985. Computer Vision, Graphics, and Image Processing 32, 29-73), or from the rearranged components of a familiar object which is itself recognizable as a 2-tone image. Even familiar volumes such as canonical bricks and cylinders require scenes with redundant structure (e.g., rows of cylinders) or explicit lighting (a lamp in the image) for recovery of global volumetric shape. We conclude that 2-tone image perception is not mediated by bottom-up extraction of geometrical features such as junctions or volumetric parts, but may rely on previously stored representations in memory and a model of the illumination of the scene. The success of this top-down strategy implies it is available for general object recognition in natural scenes. PMID- 9735537 TI - Do viewpoint-dependent mechanisms generalize across members of a class? AB - Evidence for viewpoint-specific image-based object representations have been collected almost entirely using exemplar-specific recognition tasks. Recent results, however, implicate image-based processes in more categorical tasks, for instance when objects contain qualitatively different 3D parts. Although such discriminations approximate class-level recognition. they do not establish whether image-based representations can support generalization across members of an object class. This issue is critical to any theory of recognition, in that one hallmark of human visual competence is the ability to recognize unfamiliar instances of a familiar class. The present study addresses this questions by testing whether viewpoint-specific representations for some members of a class facilitate the recognition of other members of that class. Experiment 1 demonstrates that familiarity with several members of a class of novel 3D objects generalizes in a viewpoint-dependent manner to cohort objects from the same class. Experiment 2 demonstrates that this generalization is based on the degree of familiarity and the degree of geometrical distinctiveness for particular viewpoints. Experiment 3 demonstrates that this generalization is restricted to visually-similar objects rather than all objects learned in a given context. These results support the hypothesis that image-based representations are viewpoint dependent, but that these representations generalize across members of perceptually-defined classes. More generally, these results provide evidence for a new approach to image-based recognition in which object classes are represented as cluster of visually-similar viewpoint-specific representations. PMID- 9735538 TI - Evidence accumulation in cell populations responsive to faces: an account of generalisation of recognition without mental transformations. AB - In this paper we analyse the time course of neuronal activity in temporal cortex to the sight of the head and body. Previous studies have already demonstrated the impact of view, orientation and part occlusion on individual cells. We consider the cells as a population providing evidence in the form of neuronal activity for perceptual decisions related to recognition. The time course on neural responses to stimuli provides an explanation of the variation in speed of recognition across different viewing circumstances that is seen in behavioural experiments. A simple unifying explanation of the behavioural effects is that the speed of recognition of an object depends on the rate of accumulation of activity from neurones selective for the object, evoked by a particular viewing circumstance. This in turn depends on the extent that the object has been seen previously under the particular circumstance. For any familiar object, more cells will be tuned to the configuration of the object's features present in the view or views most frequently experienced. Therefore, activity amongst the population of cells selective for the object's appearance will accumulate more slowly when the object is seen in an unusual view, orientation or size. This accounts for the increased time to recognise rotated views without the need to postulate 'mental rotation' or 'transformations' of novel views to align with neural representations of familiar views. PMID- 9735539 TI - Diagnostic recognition: task constraints, object information, and their interactions. AB - Object recognition and categorization research are both concerned with understanding how input information matches object information in memory. It is therefore surprising that these two fields have evolved independently, without much cross-fertilization. It is the main objective of this paper to lay out the basis of a dialogue between object recognition and categorization research, with the hope of raising issues that could cross-fertilize both domains. To this end, the paper develops diagnostic recognition, a framework which formulates recognition performance as an interaction of task constraints and object information. I argue and present examples suggesting that diagnostic recognition could be fruitfully applied to the understanding of everyday object recognition. Issues are raised regarding the psychological status of the interactions specified in the framework. PMID- 9735541 TI - Legionnaires' disease associated with Paris in June--update. PMID- 9735540 TI - The objects of action and perception. AB - Two major functions of the visual system are discussed and contrasted. One function of vision is the creation of an internal model or percept of the external world. Most research in object perception has concentrated on this aspect of vision. Vision also guides the control of object-directed action. In the latter case, vision directs our actions with respect to the world by transforming visual inputs into appropriate motor outputs. We argue that separate, but interactive, visual systems have evolved for the perception of objects on the one hand and the control of actions directed at those objects on the other. This 'duplex' approach to high-level vision suggests that Marrian or 'reconstructive' approaches and Gibsonian or 'purposive-animate-behaviorist' approaches need not be seen as mutually exclusive, but rather as complementary in their emphases on different aspects of visual function. PMID- 9735542 TI - HIV/AIDS mortality continues to fall. PMID- 9735543 TI - AIDS and HIV infection in the United Kingdom: monthly report. PMID- 9735544 TI - Rabies prophylaxis in Western Australia: the impact of Australian bat lyssavirus. AB - Post-exposure rabies prophylaxis is provided by the Health Department of Western Australia to persons exposed to potentially rabid animals overseas. In addition, since the discovery of Australian bat lyssavirus in 1996, rabies prophylaxis has been provided to persons exposed or likely to be exposed to Australian bats. This article reviews the provision of rabies prophylaxis in Western Australia from July 1991 to December 1997. During this period, 101 persons received rabies post exposure prophylaxis in Western Australia. Exposure occurred outside Australia in 91% of cases. Dogs were the most frequent source of exposure (62.4%) and Thailand was the most frequent country of exposure (34.7%). However in 1997, Australian bat exposures accounted for 37.5% of all post-exposure prophylaxis. No pre exposure prophylaxis was given until 1997, when eight persons received rabies vaccine to protect them against possible infection with Australian bat lyssavirus. Until the epidemiology of Australian bat lyssavirus is more clearly defined, the Lyssavirus Expert Group has recommended rabies prophylaxis be given for all Australian bat exposures. In the context of Australian bat lyssavirus as an emerging infectious disease it is important to have baseline data on rabies prophylaxis to allow for future assessment of its impact. PMID- 9735545 TI - A case of human rabies in Russia (Siberia). PMID- 9735546 TI - A case of cholera. PMID- 9735547 TI - Legionnaires' disease outbreak. PMID- 9735548 TI - Salmonellosis outbreak. PMID- 9735549 TI - Immunise Australia program: measles control campaign. PMID- 9735550 TI - Moving the second dose of measles-mumps-rubella vaccine to school entry: implications for control of rubella. PMID- 9735551 TI - Administration of measles-mumps-rubella vaccination with other childhood schedule vaccines. PMID- 9735552 TI - Changes in the reporting of HIV diagnoses in Australia. PMID- 9735553 TI - Communicable diseases surveillance. PMID- 9735554 TI - The new diagnosis and classification of diabetes mellitus. AB - We are in the midst of an epidemic of diabetes, and the prevalence appears to be especially marked within Delaware. To prevent tragic long-term complications of diabetes, and to minimize the enormous costs associated with treating them, an emphasis must be placed on the early diagnosis and aggressive management of diabetes. The changes in the classification, diagnosis and screening for diabetes should help to redirect the focus to one of preventive care. PMID- 9735555 TI - High immunization rates versus missed immunization opportunities in a private pediatric office. AB - OBJECTIVE: This prospective study examines coverage levels for immunization of two-year-old children in a large private pediatric practice, and delineates the frequency of missed opportunities for vaccination. SETTING: A private single specialty group pediatric practice in a suburban locale. DESIGN: With the aid of our office billing computer, 218 children between the ages of 21 and 24 months who had ever received medical care in our practice were identified. A manual chart review was performed to identify those children no longer receiving care from us. Of the 189 patients remaining, the immunization records in the patients' charts were compared to the State of Delaware immunization registry to identify those who had not received the full panel of recommended vaccines. Those charts were examined to identify reasons for underimmunization. No patient recall was performed. The charts of underimmunized children were reexamined five months later to determine how many had become fully immunized. RESULTS: On initial review, 86 percent of 21-24 month old children in our practice had received all recommended vaccines. Only 15 percent of the underimmunized children had had a missed opportunity for vaccination in the initial chart review. About half of the underimmunized patients received their missing vaccines over the next five months, yielding an eventual immunization rate of 94 percent. Only two of the 11 children still underimmunized by the end of the study (aged 26 to 29 months) had appeared for an 18 month or a 24 month well child visit; only four of the 11 had had a missed opportunity to receive immunizations during the five month study period. A subsequent chart review showed that 90 percent of our patients were up to-date at 24 months of age. CONCLUSIONS: Achievement of a 90 percent immunization rate for two year old children is possible in a large private practice. Most underimmunized children in our practice had failed to appear for recommended well visits; a minority experienced missed opportunities for immunization. PMID- 9735556 TI - Enhancing potency. PMID- 9735557 TI - A vision for future health care. PMID- 9735558 TI - Two-dimensional filter to facilitate detection of transient-evoked otoacoustic emissions. AB - This paper implements a filtering technique to enhance the signal-to-noise ratio (SNR) and, in turn, the detection of transient-evoked otoacoustic emissions (TEOAE's), generated by healthy human cochlea. One can increase the SNR by compiling an image of recorded TEOAE from more than one stimulus intensity, averaged over a few sweeps, which can be further processed by means of two dimensional spatial mean filters. Averaging some 60 sweeps recorded to stimuli at several intensity levels requires one-forth of the collection time needed for a classical set of responses (average of 260 sweeps), and obtains approximately the same final SNR. The relation between the performances of the proposed technique and the SNR of the rapidly acquired responses before filtering is also investigated. PMID- 9735559 TI - Effects of maternal alcohol intake on fractal properties in human fetal breathing dynamics. AB - Fractal methods have been found to be useful in characterizing biomedical signals. The use of fractal estimation requires the estimation of parameter H, which is directly related to the fractal dimension D. Here, we propose a new approach which is a combination of the wavelet transform and fractal estimators to characterize the human fetal breathing signals before and after the intake of two glasses of wine by a mother. This study was performed on 26 fetuses. The variances of the wavelet coefficients were estimated at each scale. The slope of the representation on a logarithmic plot from the scales 5 to 1 was found to be increased after alcohol intake. Our results suggested that fetal breathing rates have a rough structure before the alcohol intake and a smooth structure after alcohol intake. PMID- 9735560 TI - Denoising of the uterine EHG by an undecimated wavelet transform. AB - We propose two original methods of denoising of the uterine electrohysterography (EHG) signal by wavelets. This external electrophysiological signal is corrupted by electronic, electromagnetic noises and by the remaining electrocardiogram of the mother. The interfering signals have overlapping spectra. Therefore, a classical filtering is unusable. Wavelets should be a very well-suited denoising tool. The first proposed method uses the algorithm "a trou" with nonsymmetrical filters. The computation is rapid and the results are satisfying compared to the classical denoising techniques. The second algorithm is an improvement of the first method. It uses orthogonal wavelets and the result of the thresholding corresponds to the average of all circulant shifts denoised by a decimated wavelet transform. Results are compared to traditional denoising algorithms by wavelet (orthogonal, maximally decimated). The proposed algorithms are more efficient on simulated signals as well as on uterine EHG. PMID- 9735561 TI - The accuracy of localizing equivalent dipoles and the spatio-temporal correlations of background EEG. AB - For the inverse problem of equivalent dipole localization, a new residual function was proposed which is based on spatio-temporal correlation of background electroencephalogram (EEG). This residual has the advantage that it allows the calculation of a confidence region for estimated dipole parameters. This method was applied to two sets of visual evoked potential (VEP) data. The localization was compared by using the volume of the confidence region. The outcome of the equivalent dipole localization was compared for three different residual functions: 1) least square; 2) based on spatial correlations in the background EEG; and 3) the proposed new function which is based on spatial and temporal correlations in the background EEG. It was found that the proposed residual function leads us to the highest accuracy and the fastest convergence in the equivalent dipole localization and that even for two-dipole localization, the present method yields more accurate solutions with less iterations than the conventional methods. PMID- 9735562 TI - Neural predictive controller for insulin delivery using the subcutaneous route. AB - A neural predictive controller for closed-loop control of glucose using subcutaneous (s.c.) tissue glucose measurement and s.c. infusion of monomeric insulin analogs was developed and evaluated in a simulation study. The proposed control strategy is based on off-line system identification using neural networks (NN's) and nonlinear model predictive controller design. The system identification framework combines the concept of nonlinear autoregressive model with exogenous inputs (NARX) system representation, regularization approach for constructing radial basis function NN's, and validation methods for nonlinear systems. Numerical studies on system identification and closed-loop control of glucose were carried out using a comprehensive model of glucose regulation and a pharmacokinetic model for the absorption of monomeric insulin analogs from the s.c. depot. The system identification procedure enabled construction of a parsimonious network from the simulated data, and consequently, design of a controller using multiple-step-ahead predictions of the previously identified model. According to the simulation results, stable control is achievable in the presence of large noise levels, for unknown or variable time delays as well as for slow time variations of the controlled process. However, the control limitations due to the s.c. insulin administration makes additional action from the patient at meal time necessary. PMID- 9735563 TI - Effect of conductivity uncertainties and modeling errors on EEG source localization using a 2-D model. AB - This paper presents a sensitivity study of electroencephalography-based source localization due to errors in the head-tissue conductivities and to errors in modeling the conductivity variation inside the brain and scalp. The study is conducted using a two-dimensional (2-D) finite element model obtained from a magnetic resonance imaging (MRI) scan of a head cross section. The effect of uncertainty in the following tissues is studied: white matter, gray matter, cerebrospinal fluid (CSF), skull, and fat. The distribution of source location errors, assuming a single-dipole source model, is examined in detail for different dipole locations over the entire brain region. We also present a detailed analysis of the effect of conductivity on source localization for a four layer cylinder model and a four-layer sphere model. These two simple models provide insight into how the effect of conductivity on boundary potential translates into source location errors, and also how errors in a 2-D model compare to errors in a three-dimensional model. Results presented in this paper clearly point to the following conclusion: unless the conductivities of the head tissues and the distribution of these tissues throughout the head are modeled accurately, the goal of achieving localization accuracy to within a few millimeters is unattainable. PMID- 9735564 TI - Extracellular potentials from active myelinated fibers inside insulated and noninsulated peripheral nerve. AB - A model is presented that calculates the single-fiber extracellular field and action potential (ap) of an active myelinated nerve fiber placed centrally or eccentrically inside a nerve with a cylindrical geometry, representing essentially a one-fascicle nerve. This one-fascicle nerve has the dimensions and conductivities of the rat peroneal nerve branch. The results show a wide variety of wave shapes to be measured, depending on the position of the intraneural electrode with respect to the fiber axis and to the nodes of Ranvier and depending on the presence of an isolating cuff around the nerve. Action potential shapes may range from the "classical" quasi-biphasic one, to more triphasic, or even more complicated in the case of a short insulating cuff being present around the nerve. In the latter case, when measured bipolarly, ap-wave shapes become almost monophasic. PMID- 9735565 TI - Reduced-order modeling for hyperthermia: an extended balanced-realization-based approach. AB - Accurate thermal models are needed in hyperthermia cancer treatments for such tasks as actuator and sensor placement design, parameter estimation, and feedback temperature control. The complexity of the human body produces full-order models which are too large for effective execution of these tasks, making use of reduced order models necessary. However, standard balanced-realization (SBR)-based model reduction techniques require a priori knowledge of the particular placement of actuators and sensors for model reduction. Since placement design is intractable (computationally) on the full-order models, SBR techniques must use ad hoc placements. To alleviate this problem, an extended balanced-realization (EBR) based model-order reduction approach is presented. The new technique allows model order reduction to be performed over all possible placement designs and does not require ad hoc placement designs. It is shown that models obtained using the EBR method are more robust to intratreatment changes in the placement of the applied power field than those models obtained using the SBR method. PMID- 9735566 TI - Quantification of the 3-D electromagnetic power absorption rate in tissue during transurethral prostatic microwave thermotherapy using heat transfer model. AB - Experiments were performed in a tissue microwave-equivalent phantom gel to quantitatively examine the volumetric heating produced by a microwave antenna with a peripheral cooling system for the transurethral prostatic thermotherapy. Based on previous research, expression for the specific absorption rate (SAR) of microwave energy in the gel was extended to three dimensions, which includes its dependence on radial, angular, and axial direction. A theoretical heat transfer model was developed to study the temperature distribution in the gel by introducing this proposed SAR expression. The parameters in this expression and the convection coefficient due to the chilled water running around the antenna were determined using a least-square residual fit of the theoretical temperature predictions to the experimentally measured steady-state temperature field within the gel. The analytical expression of the three-dimensional SAR distribution obtained in this study will help provide a better understanding of the microwave heating pattern in the prostatic tissue and, thus, to aid in designing improved applicators. It can also be used in the future as an accurate input to heat transfer models which predict temperature distributions during the transurethral microwave thermotherapy. PMID- 9735567 TI - Three-dimensional reconstruction of live embryos using robotic macroscope images. AB - To determine the three-dimensional (3-D) shape of a live embryo is a technically challenging task. We show that reconstructions of live embryos can be done by collecting images from different viewing angles using a robotic macroscope, establishing point correspondences between these views by block matching, and using a new 3-D reconstruction algorithm that accommodates camera positioning errors. The algorithm assumes that the images are orthographic projections of the object and that the camera scaling factors are known. Point positions and camera errors are found simultaneously. Reconstructions of test objects and embryos show that meaningful reconstructions are possible only when camera positioning and alignment errors are accommodated since these errors can be substantial. Reconstructions of early-stage axolotl embryos were made from sets of 33 images. In a typical reconstruction, 781 points, each visible in at least three different views, were used to form 1511 triangles to represent the embryo surface. The resulting reconstruction had a mean radius of error of 0.27 pixels (1.1 microns). Mathematical properties of the reconstruction algorithm are identified and discussed. PMID- 9735568 TI - Effects of blood perfusion rate on the optimization of RF-capacitive hyperthermia. AB - The effects of the blood perfusion rate on the optimization of heating conditions in radio-frequency capacitive hyperthermia were examined using numerical simulations. When the blood perfusion rate in the tumor was smaller than approximately one-half that of normal tissues, optimal selective heating of the tumor was obtained. PMID- 9735569 TI - Stochastic complexity measures for physiological signal analysis. AB - Traditional feature extraction methods describe signals in terms of amplitude and frequency. This paper takes a paradigm shift and investigates four stochastic complexity features. Their advantages are demonstrated on synthetic and physiological signals; the latter recorded during periods of Cheyne-Stokes respiration, anesthesia, sleep, and motor-cortex investigation. PMID- 9735570 TI - Laparoscopic inguinal hernia repairs in men in a community hospital setting using the TAPP approach. PMID- 9735571 TI - Sarcoid-associated minimal change disease: a case report. AB - A patient with sarcoidosis developed minimal change disease presenting as the nephrotic syndrome. We believe this to be the first reported case of minimal change disease associated with sarcoidosis. PMID- 9735573 TI - Investing in the tort system. PMID- 9735572 TI - The supervision of medical education in South Carolina: past, present, and future. PMID- 9735574 TI - Teen pregnancy prevention: not quite a complete success story. PMID- 9735575 TI - Axis II symptomatology, depression, and life stress during the transition from adolescence to adulthood. AB - This study examined 2 models of the relationship between personality disorder symptomatology and depression, incorporating life stress as an intervening variable. In a community sample of late adolescent women, symptoms of Cluster B disorders predicted interpersonal chronic stress and self-generated episodic stress over 2 years, controlling for initial depression. Cluster A symptoms also predicted subsequent chronic interpersonal stress, over initial depression. Cluster C pathology did not predict subsequent stress. Personality disorder symptomatology was also associated with partner-reported relationship dissatisfaction. Support was found for a mediation model whereby women with higher levels of initial personality disturbance in Clusters A and B generated excessive amounts of episodic stress and interpersonal chronic stress in the next 2 years, which, in turn, increased vulnerability for depressive symptoms. A moderation model, whereby the presence of greater personality disorder symptoms would increase the likelihood of depression in response to stress, was not supported. PMID- 9735576 TI - Screening and brief intervention for high-risk college student drinkers: results from a 2-year follow-up assessment. AB - This randomized controlled trial evaluated the efficacy of a brief intervention designed to reduce the harmful consequences of heavy drinking among high-risk college students. Students screened for risk while in their senior year of high school (188 women and 160 men) were randomly assigned to receive an individualized motivational brief intervention in their freshman year of college or to a no-treatment control condition. A normative group selected from the entire screening pool provided a natural history comparison. Follow-up assessments over a 2-year period showed significant reductions in both drinking rates and harmful consequences, favoring students receiving the intervention. Although high-risk students continued to experience more alcohol problems than the natural history comparison group over the 2-year period, most showed a decline in problems over time, suggesting a developmental maturational effect. PMID- 9735577 TI - Cognitive-behavioral self-help for binge eating disorder: a controlled effectiveness study. AB - The aim of this study was to evaluate the effectiveness of 2 methods of administering a cognitive-behavioral self-help program for binge eating disorder. The study was designed to reproduce many of the conditions that apply in settings in which self-help interventions are most relevant. Seventy-two women with binge eating disorder were randomly assigned to 1 of 3 conditions for 12 weeks: pure self-help (PSH), guided self-help (GSH), or a waiting list (WL) control condition (followed by PSH or GSH). They were then followed up for 6 months. Both PSH and GSH had a substantial and sustained impact with almost half the participants ceasing to binge eat. There was little change in the WL condition. Cognitive behavioral self-help may be of value both as an initial treatment for binge eating disorder and as a form of secondary prevention. PMID- 9735578 TI - Comparison of two community alternatives to incarceration for chronic juvenile offenders. AB - The relative effectiveness of group care (GC) and multidimensional treatment foster care (MTFC) was compared in terms of their impact on criminal offending, incarceration rates, and program completion outcomes for 79 male adolescents who had histories of chronic and serious juvenile delinquency. Results show that boys who participated in MTFC had significantly fewer criminal referrals and returned to live with relatives more often. Multiple regression analyses showed that assignment to a treatment condition (i.e., GC or MTFC) predicted official and self-reported criminality in follow-up beyond other well-known predictors of chronic juvenile offending (i.e., age at 1st offense, number of previous offenses, age at referral). PMID- 9735579 TI - Therapist interventions and client emotional experiencing in expert psychodynamic interpersonal and cognitive-behavioral therapies. AB - Eighteen sessions of cognitive-behavioral (CB) and 13 sessions of psychodynamic interpersonal therapy obtained from experienced clinicians in a naturalistic setting were investigated to determine the various therapeutic interventions associated with high and low emotional experiencing (EXP). Clients receiving reflections and acknowledgments, affiliative and noncontrolling interventions, or interventions highlighting nonspecific client content were associated with maintained high EXP. Lengthier interventions and interventions rated as affiliative but moderately controlling were associated with shifts to low EXP. For clients of CB therapists, questions, interventions rated affiliative but controlling, and highlighting minimal emotional content were also associated with shifts to low EXP. Male therapists were associated with clients who maintained high EXP and female therapists were associated with clients who shifted to low EXP. PMID- 9735580 TI - Are smaller weight losses or more achievable weight loss goals better in the long term for obese patients? AB - Weight losses and psychological well-being were examined at 30 months in 69 men and 61 women initially treated with behavior therapy as a function of (a) initial weight loss and (b) weight-loss goals. Initial weight losses were positively, not negatively, related to weight loss at 30 months. Weight loss goals did not predict short-term or long-term weight loss. People who reached weight goals had better long-term weight losses than those who did not, but this finding was largely due to differences in initial weight loss. Psychological well-being at 30 months was not related to initial weight losses or goals. Although correlational rather than experimental, these results do not support the hypothesis that obese patients should be encouraged to set lower weight-loss goals. PMID- 9735581 TI - Differential relation of psychological functioning with the history and experience of weight cycling. AB - Two measures of weight cycling and indexes of psychological functioning were examined in a large sample of dieters. History of weight cycling was assessed to include number of dieting attempts, total lifetime weight lost and regained, and number of weight cycles over 20 lb (9.1 kg). Experience of weight cycling measured perception of being a yo-yo dieter and perceived success at maintaining past weight losses. Experience was more strongly related than history to all psychological measures. Further, when controlling for the effects of age, body mass index, and experience, the relation between history and the psychological variables was nonsignificant. This finding suggests that an individual's perception of being a weight cycler may be more related to psychological problems than the actual number of pounds lost and regained over time. PMID- 9735582 TI - Assessment of body image dissatisfaction in obese women: specificity, severity, and clinical significance. AB - This study assessed the specificity, severity, and clinical significance of body image dissatisfaction in 79 obese women using the Body Dysmorphic Disorder Examination--Self-Report (J. C. Rosen & J. Reiter, 1996). The vast majority of obese women demonstrated body image dissatisfaction related to their obesity, with almost half reporting the greatest dissatisfaction with their waist or abdomen. On average, they reported significantly more body image dissatisfaction than did 43 nonobese controls. The 2 groups did not differ on self-reported symptoms of depression or self-esteem. Body image dissatisfaction correlated significantly with reports of depressive symptoms and lower self-esteem but was not correlated with body mass index. Results are discussed in terms of the role of body image dissatisfaction in understanding and treating obese individuals. PMID- 9735583 TI - Does the gender of a patient or the gender of a therapist affect the treatment of patients with major depression? AB - The role of gender was examined in the process and outcome of therapy in the treatment of depressed outpatients seen in the National Institute of Mental Health Treatment of Depression Collaborative Research Program. Patients received either interpersonal therapy, cognitive-behavioral therapy, imipramine plus clinical management, or placebo plus clinical management. None of the therapist patient by gender groupings (i.e., therapist gender, therapist-patient gender matching vs. mismatching, or patients' beliefs about whether a male or female therapist would be more helpful) were significantly related to measures of treatment process and outcome, controlling for type of treatment and severity of pretreatment depressive symptoms. Findings were duplicated when examining the effects of gender within only the psychotherapeutic modes of treatment for the groupings of therapist gender and therapist-patient gender matching versus mismatching. PMID- 9735584 TI - Diagnosing attention deficit disorders with the Behavioral Assessment System for Children and the Child Behavior Checklist: test and construct validity analyses using optimal discriminant classification trees. AB - The usefulness of the Behavioral Assessment System for Children (BASC) and Child Behavior Checklist (CBCL) Parent scales was examined with respect to (a) differentiating students with attention deficit-hyperactivity disorder (ADHD) from non-ADHD students and (b) discriminating between the predominantly inattentive-type and combined-type ADHD-afflicted students. For both the BASC and the CBCL, a different optimal discriminant classification tree analysis (CTA) model was developed for each of the 2 diagnostic predictions. For distinguishing ADHD students from non-ADHD students, the BASC model was more parsimonious and accurate than the CBCL model. Toward the goal of differentiating between primarily inattentive and combined types, the CBCL's model was superior for predicting primarily inattentive students. The results demonstrate the diagnostic utility of the BASC and CBCL and describe salient behavioral dimensions associated with subtypes of ADHD. PMID- 9735585 TI - Cognitive factors differentiating attention deficit-hyperactivity disorder with and without a comorbid mood disorder. AB - Mood disorders and attention deficit-hyperactivity disorder (ADHD) co-occur in 20 30% of children and adolescents diagnosed in both epidemiological and clinical studies, but little information is available regarding cognitive factors that may be relevant to the expression of co-occurring mood disorders and ADHD. This study examined whether ADHD with and without a comorbid mood disorder could be differentiated on the basis of cognitive factors associated with prominent theories of depression. Children meeting diagnostic criteria for ADHD (n = 14) or ADHD and a comorbid mood disorder (n = 27) were assessed on a variety of cognitive indices. Children in the comorbid group reported more negative views of themselves and a more depressogenic attributional style. Cognitive disturbances associated with A. T. Beck's (1967) cognitive model and attributional style theories of depression differentiate ADHD children with significant mood pathology. PMID- 9735586 TI - Increasing assertiveness skills to reduce HIV risk among women living with a severe and persistent mental illness. AB - This study examined whether training women living with a severe mental illness to be assertive in sexual situations would decrease their risk for HIV infection. Twenty female outpatients were randomly assigned to either a 10-session assertiveness training intervention or a waiting-list control condition. All participants completed measures of HIV-related information, motivation, skills, and sexual risk behavior pre- and postintervention and at 2- and 4-month follow ups. Compared with controls, women in the intervention group increased their assertiveness skill, HIV knowledge, and frequency of condom-protected intercourse. It is concluded that assertiveness training for women living with a severe mental illness can serve as 1 part of a comprehensive HIV-risk-reduction program for this vulnerable population. PMID- 9735587 TI - Cognitive bibliotherapy for mild and moderate adolescent depressive symptomatology. AB - The efficacy of cognitive bibliotherapy for adolescents experiencing mild and moderate depressive symptomatology was examined with a group of 22 community dwelling adolescents. Cognitive bibliotherapy was determined to be superior to a delayed-treatment control condition. The treatment produced both statistically and clinically significant improvements in depressive symptoms. Treatment gains were maintained at 1-month follow-up. A significant decrease in dysfunctional thoughts, but not in negative automatic thoughts, was found after treatment. These results contribute to converging evidence on the efficacy of cognitive behavioral treatments for adolescents experiencing depressive symptoms. PMID- 9735588 TI - Cocaine use early in treatment predicts outcome in a behavioral treatment program. AB - In this evaluation of baseline drug use as a predictor of treatment outcome, cocaine use during a 5-week baseline was compared in methadone maintenance patients who had < 5 (n = 10) versus > or = 5 (n = 9) weeks of abstinence during an experimental cocaine abstinence reinforcement treatment. Cocaine use was evaluated at the 1st and last visit and the 1st and last week of baseline and as a mean across the 5-week baseline treatment; response was calculated as a mean across 12 weeks of experimental treatment. Those who had successful outcomes (abstainers) used significantly less cocaine in the 5-week baseline than those with less successful outcomes (nonabstainers). Differences in cocaine use were not evident in the 1st baseline visit or week, but the abstainers used significantly less cocaine in the last visit and week of baseline compared with the nonabstainers. Cocaine use during baseline provided critical predictors of response to the experimental treatment. PMID- 9735589 TI - Random versus nonrandom assignment in the evaluation of treatment for cocaine abusers. AB - Cocaine-abusing patients randomly assigned to day-hospital or inpatient rehabilitation were compared with patients who self-selected these treatment settings to examine differences in substance use and psychosocial outcomes under experimental and nonexperimental designs. There was little evidence of setting or assignment effects or Setting x Assignment interactions over the 12-month follow up period. However, Assignment x Time interactions were obtained with 2 measures of cocaine use and measures of family-social and psychiatric problem severity. These interactions indicated greater problem severity at intake among the randomized patients coupled with greater improvements by the 3-month follow-up relative to the nonrandomized patients. Findings suggest that randomized studies of treatment for cocaine abuse may produce somewhat larger estimates of improvement than what is observed in more typical treatment situations. PMID- 9735590 TI - Bibliotherapy in the treatment of sexual dysfunctions: a meta-analysis. AB - This article describes the combined effect of 12 controlled studies of bibliotherapy for sexual dysfunctions, comprising data on 397 participants, who were treated in 16 bibliotherapy groups. A mean effect size of 0.68 SDs at posttreatment was found (0.50 when weighted for sample size). This effect eroded at follow-up. No influence on effect size was found for either bibliotherapy implementation characteristics or study methodology. Studies were largely limited to bibliotherapeutic administration of the directed practice approach to orgasmic disorders. The efficacy of bibliotherapy has not yet been investigated sufficiently for evaluation of its use for other sexual dysfunctions or for its comparison with other therapeutic approaches for sexual dysfunctions. PMID- 9735591 TI - Human work capacity under combined stress of work and heat. AB - The working capacity of young, healthy, unacclimatized men (N = 11) was studied under long-duration (8 to 9 days) exposure to combined work and heat (dry and humid). The dry (Gr A, N = 5) and humid (Gr B, N = 6) groups were exposed to 41.3 +/- 0.6 degrees C DB, 40-50% RH and 39.2 +/- 0.6 degrees C DB, 70-80% RH, respectively, for all days of exposure. The experimental protocol was divided into: (i) direct determination of maximal oxygen uptake (VO2max) by stepped increases in bicycle ergometry everyday in the morning in the initial hours before exposure to heat, after which the recovery process of oxygen debt contraction was examined; and (ii) exposure to heat in a climatic chamber for 2 h where the subjects performed two spells of ergometric work (10 to 12 min each) at a relative intensity of 50 +/- 12 to 69 +/- 11% VO2max. The average heat exposure time for Gr A was higher (108 +/- 12 min) as compared to Gr B (95 +/- 10 min), but Gr B sustained a high heat load as reflected from the high deep-body temperature maintained during the exposure. The high body temperature load of Gr B had a significant effect on the cardiorespiratory capacity, indicating an upward trend in VO2max. This was statistically significant (p < 0.05) for the first four days of exposure. Subjects of Gr B had a relatively higher working capacity compared to those in Gr A on all days. The VO2max and analysis of the fractions of oxygen debt contraction (fast and slow component) indicated that the subjects showed a better training/heat acclimatization effect under hot, humid conditions. PMID- 9735593 TI - Negative affectivity influences report of work-related symptoms. AB - A number of studies have documented that negative affectivity (NA) exerts a considerable influence upon perceptual style and report of symptoms. In three studies (N = 24, N = 30, N = 43), the current paper investigated the influence of NA upon the report of work-related symptoms. For all three studies, we found a firm association between NA and visual and muscular-skeletal symptoms. The results are discussed according to pain sensitivity, general activation and symptom perception. The paper concludes that NA might be a methodological nuisance factor leading to the over-reporting of work-related complaints. PMID- 9735592 TI - The effect of wrist and arm postures on peak pinch strength. AB - This paper examines the effect of various arm postures on peak pinch strength. Twenty (20) able-bodied, male subjects volunteered to participate in a set of two experiments. The first experiment examined the effect of shoulder and elbow posture on peak pinch strength. The second experiment examined the effect of forearm posture, wrist posture, and pinch type on peak pinch strength. Results from the first experiment indicated that elbow posture had a significant effect on pinch strength. It was documented that extreme elbow flexion decreased pinch strength by up to 9%. Results from the second experiment indicated that deviated wrist postures, forearm postures, and pinch type significantly decreased pinch strength as much as 33%. Ergonomic guidelines which utilize the above factors as significant modifiers of pinch strength capacity may assist ergonomists to reduce the risk of injury and development of cumulative trauma disorders in the workplace. PMID- 9735594 TI - Some design recommendations to improve comfort in helmets: a case study from China. AB - Unless the basic user needs are satisfied in safety helmets, it is difficult to get workers to wear them habitually and for long periods. Hotness, weight and fitting problems are major wearability issues that require improvements. The enormous need for an optimally designed helmet in China prompted a case study on comfort aspects in helmets. The subjective impressions of the wearers of test helmets provided useful information for design changes to improve comfort. The heat transfer measurements through helmets indicated the need for ventilation openings to be provided on the shell of plastic helmets. Due to the advantage of low weight and good ventilation, it is recommended that cane helmets be further developed to improve protection, wearability and durability, and subsequently be produced in large scale. PMID- 9735595 TI - Random errors in anthropometry. AB - In order to present basic information on the magnitude of and variance due to the random error in anthropometry, 219 measurement items were taken on 12 subjects twice by the same observer. The precision (i.e., consistency between the repeated measurements) was investigated for these measurement items. The reliability was quantified using mean absolute difference (MAD), technical error of measurement (TEM), and reliability coefficient (R). MAD and TEM are highly correlated with each other and both represent the magnitude of error. They are not correlated with R, which represents the proportion of error-free variance. Larger measurements tend to have absolutely larger but relatively smaller random errors and higher reliability in the size range of 1-10 cm. Imprecision is inherent in anthropometry of the living because of the fact that the human body is not rigid. This may be responsible for the above tendency. Relatively large MAD and low R may be due to small absolute size, landmarks difficult to locate precisely, soft tissue deformation, and the inconsistency of the posture of the subject. PMID- 9735596 TI - Fundamental study on the size and inter-key spacing of numeric keys for touch screen. AB - The purpose of this study was to reveal the optimum size and inter-key spacing of numeric square keys for touch screens. Six male students (22-25 years old) and three female students (21-24 years old) volunteered as subjects for this experiment. Each subject took part in data entry tasks using numeric square keys of touch devices. The sizes of keys were 6, 12, 21, 30 and 39 mm and each the inter-key spacing was 0, 3, 6, 12 and 21 mm. Response times with key sizes of 6 and 12 mm were significantly slower than with key sizes of 21 and 30 mm (p < 0.001). Furthermore, the key size of 6 mm significantly caused more errors than the key sizes of 12, 21, 30 and 39 mm (p < 0.05). The response time with inter key spacing of 3 mm was significantly faster than with that of 0, 6, 12 and 21 mm (p < 0.001). Inter-key spacing of 0 mm significantly produced more errors than other inter-key spacing. Subjective ratings for inter-key spacing of 3, 6 and 12 mm were significantly better than those of 0 and 21 mm (p < 0.05). These results suggested that the optimum size of numeric square keys for touch screens should be more than 21 mm and optimum inter-key spacing should be from 3 to 6 mm. Optimum key size, however, must be selected with regard to the limitation of screen size. PMID- 9735597 TI - Comparison of tumorigenesis between accelerated heavy ion and X-ray in B6C3F1 mice. AB - The effects of heavy ion and X-ray irradiation on tumorigenesis in B6C3F1 mice were compared. Six-week-old animals were divided into 6 groups and exposed to 0.426 Gy heavy ion irradiation of 290 MeV/u carbon-ion beam (LET 60-210 KeV/micron) at the dose rate of 0.4 +/- 0.2 Gy/min; 0.5 Gy of X-ray irradiation at 0.1 Gy/min or 5 Gy of X-ray irradiation at 1 Gy/min. The mice were killed and an autopsy performed 13.5 months after the whole body irradiation. Body weights were heaviest for both sexes in the 0.5 Gy group and lightest in the 5 Gy one. Total tumor incidences in the males were 30, 56 and 13% respectively in the heavy ion, 5 Gy and 0.5 Gy X-irradiated groups, stomach tumors, lymphomas and adrenal tumors being the most common outcome of the high dose X-rays. Liver tumor induction did not differ significantly among the groups. In the females tumorigenicity was significantly lower for heavy ion than for 0.5 Gy and 5 Gy X ray irradiation (P < 0.05), the respective incidences, mainly ovary one, being 73%, 17% and 41%. Non-cancerous lesions, such as graying of the hair, glomerular sclerosis and amyloidosis appeared in the 5 Gy group. These findings indicate that 0.426 Gy of heavy ion irradiation induced lower carcinogenicity than 5 Gy of X-irradiation and higher carcinogenicity than that of 0.5 Gy X-irradiation in male mice. PMID- 9735598 TI - Effect of the combination of a local OK-432 injection and hyperthermia on SCC VII tumors in mice. AB - Hyperthermia is being investigated as a cancer treatment. Many of its basic mechanisms, particularly those related to cell killing, are still poorly understood. We used a transplanted squamous cell carcinoma cell line to investigate the therapeutic effect of hyperthermia. In particular, we examined the effect of OK-432 (biological response modifier) on hyperthermia-induced apoptosis. In the hyperthermia only group the most extensive necrosis occurred on day 3 (70.3%), and the apoptosis index also was highest on that day (69.3). These results suggest that the induction of apoptosis is closely related to the cell death caused by hyperthermia. The percent of necrosis was significantly higher in the groups given hyperthermia and combined OK-432 and hyperthermia treatment than in the OK-432 group (p < 0.05). The apoptosis index was significantly lower in the combined OK-432 and hyperthermia treatment group than in the hyperthermia only group, indicative that the antitumor effect of combined hyperthermia and OK 432 therapy is not ascribable to the induction of apoptosis. PMID- 9735599 TI - Sensitivity to ionizing radiation in Saos-2 cells transfected with mutant p53 genes depends on the mutation position. AB - We have constructed an in vitro system to examine how p53 mutants affect radiosensitivity. Mutations of p53 were made using in vitro mutagenesis, and mutant cDNAs were introduced into the human osteosarcoma cell line, Saos-2, which is devoid of endogenous p53. For wild type p53, both the expression plasmid and a regulation plasmid (LacSwitch system) were transfected into the cells. The radiosensitivities of clones of mutant p53 and wild type p53 were examined. Transformants of wild type p53 had increased radiosensitivity. The induction of wild type p53 protein by addition of IPTG did not significantly increased radiosensitivity. A mutation at codon 123 also increased radiosensitivity. Mutations at codons 143, 175, and 273 did not alter radiosensitivity. PMID- 9735600 TI - A comparison of UVB-carcinogenesis between nude mice and nude beige mice. AB - To gain an insight into the relationship between UVB-carcinogenesis and natural killer activity, we examined ultraviolet light-induced carcinogenesis in mice with high natural killer, activity (KSN) and mice with natural killer deficiency (KSN-bg). We exposed mice six times a week to three levels of daily ultraviolet B (UVB) doses; 320, 160 and 0 J/m2/day. During the latency period of skin tumor development in KSN mice, we detected no suppression of the natural killer activity at both 320 and 160 J/m2/day. Even at 1340 J/m2/day, we could not detect any significant suppression of NK activity in KSN mice. When we irradiated spleen cells in vitro, we observed NK activity suppression. Next, we compared the carcinogenic effects of UVB-irradiation on KSN and KSN-bg mice. At 320 J/m2/day, we detected no significant differences between them. In contrast, at 160 J/m2/day, KSN-bg mice showed a significantly higher rate of skin tumor induction than KSN mice (p < 0.05). Most UVB-induced tumors were squamous cell carcinoma, the rest were spindle cell carcinoma, papilloma and mixed type. Our results suggest that NK activity plays a protective role against UVB-carcinogenesis from low daily-doses of UVB-irradiation. PMID- 9735601 TI - Tritium concentrations in pine needle, litter and soil samples. AB - Samples of pine needle, litter and soil samples collected in/around Akita City and Rokkasho Village in 1989 were analyzed for both free water 3H (FWT) and organically-bound 3H (OBT). The FWT concentrations decrease in the order, litter or soil > pine needle. FWT concentrations in soil depend on the moisture content, and tend to increase with decreasing soil moisture content. This relationship is consistent with the observation that FWT in the soil increases with oxidation of atmospheric tritiated hydrogen gas (HT) and decreases with rainwater dilution. The OBT concentrations increase in the order pine needle < litter < soil at most of the sampling locations. This suggests that historically high soil 3H concentrations may be reflected as high OBT concentrations in soils of the present. PMID- 9735602 TI - 3'-blocking damage of DNA as a mutagenic lesion caused by hydrogen peroxide in Escherichia coli. AB - Ionizing radiation and hydrogen peroxide (H2O2) produce many types of oxidative DNA damage such as strand breaks, apurinic/apyrimidinic (AP) sites, base modifications and 3'-blocking damage such as 3'-phosphoglycolated and 3' phosphorylated termini. AP sites and 3'-blocking damage are repairable by exonuclease III and endonuclease IV in Escherichia coli. XthA-nfo double mutants of E. coli, which are deficient in exonuclease III and endonuclease IV, were highly sensitive to lethal and mutagenic effects of H2O2, compared with the wild type strains. The pNT180 and pNT186 plasmids containing wild-type nfo and mutant nfo-186 gene, respectively, were introduced into the xthA-nfo mutant. The nfo-186 gene product, Nfo186, retained normal AP endonuclease activity but could not remove 3'-blocking damage from DNA. The pNT180 corrected the sensitivity of the xthA-nfo mutant to lethal and mutagenic effects of H2O2. On the other hand, the pNT186 did not have any complementation effects. From these results it was concluded that 3'-blocking damage rather than an AP site is the primary lesion responsible for both lethal and mutagenic effects of H2O2. PMID- 9735603 TI - Lissencephaly revisited. PMID- 9735604 TI - Overzealous prescribing of medications. PMID- 9735605 TI - Fluoxetine treatment in a 2.5-year-old girl. PMID- 9735606 TI - Olanzapine for autistic disorder with hyperactivity. PMID- 9735607 TI - Disruptive disorders, thyrotoxicosis, and DSM-IV. PMID- 9735608 TI - Multimodal intervention for selective mutism in mentally retarded children. PMID- 9735609 TI - Psychotherapy in eastern Europe. PMID- 9735610 TI - Predictors of treatment efficacy in a clinical trial of three psychosocial treatments for adolescent depression. AB - OBJECTIVE: To assess the predictors of treatment outcome across treatments, as well as those associated with differential treatment response. METHOD: One hundred seven adolescent outpatients, aged 13 to 18 years, with DSM-III-R major depression were randomly assigned to one of three manual-based, brief (12 to 16 sessions) psychosocial treatments: cognitive-behavioral therapy (CBT), systemic behavioral family therapy, or nondirective supportive therapy. Those with good and poor outcomes were compared. RESULTS: Continued depression was predicted by clinical referral (versus via advertisement) and was in part mediated by hopelessness. Other predictors of depression were comorbid anxiety disorder and higher levels of cognitive distortion and hopelessness at intake. Achievement of clinical remission was predicted by a higher level of self-reported depression. Poorer functional status was predicted by a higher level of initial interviewer rated depression. Comorbid anxiety and maternal depressive symptoms predicted differential treatment efficacy. CBT's performance continued to be robust with respect to nondirective supportive therapy, even in the presence of the above noted adverse predictors. CONCLUSION: Predictors of poor outcome may give clues as to how to boost treatment response. Subjects who come to treatment for clinical trials via advertisement (versus clinical referral) may show more favorable treatment responses. CBT is likely to be a robust intervention even in more complex and difficult-to-treat patients. PMID- 9735611 TI - Psychopathology associated with suicidal ideation and attempts among children and adolescents. AB - OBJECTIVE: To identify the independent and differential diagnostic and symptom correlates of suicidal ideation and suicide attempts and determine whether there are gender- and age-specific diagnostic profiles. METHOD: The relationships between suicidal ideation, suicide attempts, and psychiatric disorders were examined among 1,285 randomly selected children and adolescents, aged 9 to 17 years, of whom 42 had attempted suicide and 67 had expressed suicidal ideation only. Youths and their parents were interviewed as part of the Methods for the Epidemiology of Child and Adolescent Mental Disorders (MECA) Study, using the Diagnostic Interview Schedule for Children Version 2.3 (DISC-2.3). RESULTS: Logistic regression analyses indicated that mood, anxiety, and substance abuse/dependence disorders independently increased the risk of suicide attempts, after controlling for sociodemographic characteristics. There was no significant independent contribution of disruptive disorders to suicide attempts, although its association with suicidal ideation was significant. Substance abuse/dependence independently differentiated suicide attempters from ideators. Noncriterion symptoms that remained significant predictors of suicide risk, after adjusting for psychiatric disorder, included panic attacks and aggressiveness. Perfectionism did not significantly increase suicide risk after adjusting for psychiatric disorder. The association of specific disorders and noncriterion symptoms with suicidality varied as a function of gender and age. CONCLUSION: A monolithic diagnostic risk profile for suicidality, ignoring gender- and age specific risks, is inadequate. The contribution of substance abuse/dependence in the escalation from suicidal thoughts to suicide attempts is underscored. PMID- 9735612 TI - Psychiatric diagnoses of previous suicide attempters, first-time attempters, and repeat attempters on an adolescent inpatient psychiatry unit. AB - OBJECTIVE: To compare psychiatric diagnoses of hospitalized adolescents who (a) have made previous but no recent suicide attempts, (b) have recently made their first suicide attempt, (c) have recently made a second or subsequent attempt, or (d) have never made an attempt. METHOD: Semistructured psychiatric diagnostic interviews were used to determine psychiatric diagnoses and history of recent and previous suicidal behavior of 269 consecutively admitted adolescents to an inpatient psychiatric facility. Forty-nine previously suicidal youths, 28 first time attempters, and 33 repeat attempters were compared with 159 nonsuicidal youths in prevalence of Axis I psychiatric disorders and psychiatric comorbidity with affective disorder. RESULTS: Previous attempters and repeat attempters both reported more affective disorders, whereas first-time attempters reported more adjustment disorders than nonsuicidal youths. Previous attempters and nonsuicidal youths reported the most externalizing disorders. CONCLUSIONS: Previous attempters on an inpatient unit have multiple psychiatric problems. Like repeat attempters, they often are depressed, but like nonsuicidal youths, they also exhibit significant externalizing behaviors. Interventions with these adolescents should focus not only on immediate presenting problems, but also on ameliorating their long-term risk of posthospitalization suicidal behavior. PMID- 9735613 TI - Relationship of asthma severity and psychological problems in children. AB - OBJECTIVE: To determine whether physiological severity of asthma is associated with increased psychological symptoms in children. METHOD: Participants were 337 children, aged 7 to 19 years (mean 11.9, SE 0.13), and a parent of each child. Children's asthma severity was rated by experienced pediatric asthma specialists using current guidelines from the National Heart, Lung, and Blood Institute. Children filled out the Children's Manifest Anxiety Scale and the Weinberger Adjustment Inventory. Parents reported on their child's medical history, completed the Child Behavior Checklist (CBCL) about their child, and completed the Pennebaker Inventory of Linguid Languidness as a measure of their own physical symptoms. RESULTS: Child-rated anxiety symptoms were unrelated to asthma severity or to markers of asthma functional morbidity. Parental ratings of internalizing symptoms in their children were related to severity. Parent physical symptoms explained 10.2% of the variance in CBCL Internalizing symptoms, and asthma severity added an additional 6.7% to the variance. CONCLUSIONS: Asthma severity may be a more salient stressor to parents, who in turn report higher levels of child internalizing symptoms for children with severe asthma, than to children themselves. Contrary to prior hypotheses, children with severe asthma did not rate themselves as having higher levels of anxiety than those with mild or moderate asthma or than standardized norms. PMID- 9735614 TI - Headaches and psychopathology in children and adolescents. AB - OBJECTIVE: To examine the association between chronic headaches and DSM-III-R defined psychiatric disorders, including depression, anxiety disorders, conduct disorder, oppositional defiant disorder and attention-deficit hyperactivity disorder, in a population-based sample of children and adolescents. METHOD: 1,013 children aged 9 to 15 years in the Great Smoky Mountains Study were evaluated annually over a 3-year period using the Child and Adolescent Psychiatric Assessment, a child and parent diagnostic psychiatric interview. Headaches that lasted at least 1 hour and occurred at least once a week during the 3 months prior to the interview were studied. RESULTS: Girls with depression and anxiety disorders had a significantly greater prevalence of headaches than girls without an internalizing disorder. This association was not found for boys. Conduct disorder was significantly associated with headaches in boys. Each of these associations was constant with age. CONCLUSIONS: This study suggests that a distinct gender difference exists between boys and girls in the associations between headaches and psychopathology. Carroll's theory of dysfunction in central pain regulation as an underlying cause of depression is discussed in relation to the proposed serotonergic dysregulation common to headaches, depression, anxiety, aggression, and pain. PMID- 9735615 TI - Diagnosing infantile anorexia: the observation of mother-infant interactions. AB - OBJECTIVE: This study has three objectives: (1) to delineate the diagnostic criteria for infantile anorexia, including the onset of persistent food refusal during the infant's transition to spoon- and self-feeding, acute and/or chronic malnutrition, parental concern about the infant's poor food intake, and mother infant conflict, talk, and distraction during feeding; (2) to determine the interrater agreement of child psychiatrists when diagnosing infantile anorexia based on these criteria; and (3) to describe the use of the Feeding Scale as a diagnostic tool. METHOD: One hundred two toddlers, ranging in age from 12 to 37 months, were assessed by two child psychiatrists and assigned the diagnosis of infantile anorexia, picky eater, or good eater. In addition, observers who were masked to the toddler's diagnosis rated mother-infant interactions with the Feeding Scale to permit objective evaluation of those interactions. RESULTS: Two child psychiatrists were able to assign toddlers to infantile anorexia, picky eating, and healthy, good eating groups with a high level of agreement. The objective scale for rating mother-infant interactions showed a high level of agreement between two masked raters and a good level of agreement between masked raters and the child psychiatrists' diagnostic assessment. CONCLUSIONS: Infantile anorexia can be diagnosed with high reliability by child psychiatrists. Evaluation of mother-infant interactions is a useful diagnostic tool. PMID- 9735616 TI - Variations in ADHD treatment among special education students. AB - OBJECTIVE: To examine variation in patient characteristics (case-mix) and treatment patterns for attention-deficit/hyperactivity disorder (ADHD) by provider type. METHOD: By means of a two-stage study design, 102 children were identified as receiving treatment for ADHD in the past year, among a school district-wide sample of second-through fourth-grade special education students. Parent and child interviews were conducted using standardized measures of need for treatment, service use, and process of care. RESULTS: Nearly three fourths of the children received treatment for ADHD by a primary care provider, and of these, 68% did not have any contact with a mental health specialist. Children treated only by a primary care provider had fewer comorbid conditions, less impairment, and lower levels of family burden than children treated only by a mental health specialist. Treatment of ADHD solely by primary care providers was characterized by fewer sessions, less time with the patient, and less use of multimodal therapies. CONCLUSION: Future studies examining clinical outcomes for ADHD treatment should take into account variation in case-mix and treatment patterns by provider type. PMID- 9735617 TI - Associations between event-related potentials and measures of attention and inhibition in the Continuous Performance Task in children with ADHD and normal controls. AB - OBJECTIVES: First, to differentiate between inattention and impulsivity based on type of errors made in the AX version of the Continuous Performance Task (CPT), and second, to investigate whether differences in performance between children with attention-deficit hyperactivity disorder (ADHD) and normal controls also occur in specific forms of brain activity, namely event-related potentials (ERPs), presumably related to inattention and impulsivity or inhibition. METHOD: Sixteen ADHD and 16 normal control children performed the CPT-AX. ERPs were recorded at occipital (Oz), parietal (Pz), central (Cz), and frontal (Fz) leads. RESULTS: The ADHD children had a higher CPT-Inattention score and showed smaller parietal positive waves at a latency of approximately 300 msec in reaction to target stimuli, target P3s, likewise indicating less attention. In contrast, they showed neither higher CPT-Impulsivity nor a smaller frontocentral negative wave at about 200 msec (N2); the N2 is generally seen as reflecting inhibition. A subgroup of children with ADHD and oppositional defiant disorder (n = 6) had smaller N2 waves than controls, however. CONCLUSIONS: The ADHD group studied showed deficits in attention but not in impulsivity (or inhibition). PMID- 9735618 TI - Case study: dawn simulation as maintenance treatment in a nine-year-old patient with seasonal affective disorder. AB - After four winter seasons of successful treatment with light boxes, a 9-year-old patient with seasonal affective disorder refused to make further use of the light box. Instead he was treated with dawn simulation (a dim light administered just before waking up and gradually increased in intensity). The patient used dawn simulation therapy from October until mid-May, with an occasional variation of the maximum light intensity (100, 200, or 300 iux). The patient, his parents, and his teacher were all happy with this type of treatment. PMID- 9735619 TI - When child and adolescent psychiatrists conduct managed care reviews. PMID- 9735620 TI - Development of the cerebral cortex: IX. Cortical development and experience: I. PMID- 9735621 TI - Summary of the practice parameters for the assessment and treatment of children and adolescents with posttraumatic stress disorder. American Academy of Child and Adolescent Psychiatry. AB - This summary provides an overview of the assessment and treatment recommendations contained in the Practice Parameters for the Assessment and Treatment of Children and Adolescents With Posttraumatic Stress Disorder. Major recommendations include the use of clinical interviewing with specific questioning about posttraumatic stress symptoms to diagnose this disorder; recognition of developmental considerations that may impact on how posttraumatic stress disorder symptoms manifest in children; and the use of trauma-focused treatment interventions. Limitations and controversies regarding the present state of knowledge in the area of childhood posttraumatic stress disorder are also discussed. PMID- 9735622 TI - Foot and ankle injuries related to rock climbing. The role of footwear. AB - Interest in rock climbing has grown dramatically over the past decade. Although considerable research has been conducted on upper-extremity injuries sustained during rock climbing, there has been no comprehensive evaluation of lower extremity injuries and related biomechanics. The authors performed a retrospective investigation of rock-climbing injuries using a survey of 104 active rock climbers of varying levels of expertise. The results show that 81% of the respondents have suffered acute or chronic pain or associated pathology in the foot or ankle during or after climbing. The authors propose that this morbidity has biomechanical etiologies related to the common practice among rock climbers of wearing climbing shoes that are smaller than their street shoes. PMID- 9735623 TI - Conservative treatment of plantar fasciitis. A prospective study. AB - A randomized, prospective study was conducted to compare the individual effectiveness of three types of conservative therapy in the treatment of plantar fasciitis. One hundred three subjects were randomly assigned to one of three treatment categories: anti-inflammatory, accommodative, or mechanical. Subjects were treated for 3 months, with follow-up visits at 2, 4, 6, and 12 weeks. For the 85 patients who completed the study, a statistically significant difference was noted between groups, with mechanical treatment with taping and orthoses proving to be more effective than either anti-inflammatory or accommodative modalities. PMID- 9735624 TI - The reliability of three techniques for measuring foot position. AB - The reliability of three commonly used techniques for measuring foot position- valgus index, navicular height, and arch height--was evaluated in a study involving 20 healthy subjects. The results demonstrated significant differences (P < .05) between two observers for all three techniques, although there were no significant differences between two visits for the same observer (P < .05). Secondary analysis demonstrated that navicular height yielded the highest degree of intraobserver and interobserver agreement. The results suggest that there is a wide variation in foot position in the general population, and that measurement error may result from difficulties in defining foot position, techniques used, and instrumentation. PMID- 9735625 TI - Magnetic resonance imaging of ankle ligament injuries correlated with time. AB - Ankle sprain is one of the most commonly treated injuries of the lower extremity. The treatment depends on the severity of the injury and the time at which it occurred. The physician must rely on the history as related by the patient to determine the age of the injury. Magnetic resonance imaging has been proven to help determine the severity of the injury but has not been used to determine the age of the injury. The present study was conducted to identify the typical findings of acute and chronic ankle sprains as a means of dating an ankle sprain based on its appearance on magnetic resonance imaging. PMID- 9735626 TI - Closed rupture of the anterior tibial tendon. A case report. AB - Closed subcutaneous rupture of the anterior tibial tendon is a relatively uncommon injury that requires a thorough clinical examination to diagnose correctly. The authors report a case of this disorder and provide a review of the relevant literature. A method of surgical repair not previously described in the literature is also presented. PMID- 9735627 TI - Metastatic breast cancer presenting as heel pain. AB - The authors present a case of breast cancer metastasizing to the calcaneus that was confirmed by bone biopsy. The patient's complaint of heel pain provided the initial evidence of skeletal metastasis. Metastatic spread of cancer to the hand or foot (acrometastasis) is considered rare. However, the possibility of acrometastasis should be considered in any patient with a history of cancer presenting with skeletal pain, especially if the symptoms do not respond to therapy. PMID- 9735628 TI - Refsum's disease. A unique case. AB - Refsum's disease, or heredopathia atactica polyneuritiformis, is a peroxisomal disorder leading to the accumulation of phytanic acid throughout the body. It affects sensory and motor neurons and the skeletal system. Peripheral neuropathy, ataxia, blindness, deafness, and skeletal hyperostosis are significant findings used in the diagnosis of the disease. PMID- 9735629 TI - Bilaterally symmetrical epidermal inclusion cysts with foreign-body giant-cell reaction. AB - Epidermal inclusion cysts often occur as a result of traumatic implantation of epidermal cells into dermal tissue. The epidermal cells within the dermis can continue to grow and lead to the production of a lipid- and keratin-filled cyst, which can erode into bone and adjacent tissues. The authors present a case of bilaterally symmetrical epidermal inclusion cysts that occurred separately over a 10-year period. A brief review of the literature is also presented. PMID- 9735630 TI - Random musing about a little blue pill. PMID- 9735631 TI - AFMC project saves Arkansas Medicaid dollars & improves quality of care for women. PMID- 9735632 TI - AFMC solves the Medicare puzzle. PMID- 9735633 TI - Malpractice claims result more from neurologically impaired newborn cases than any other reason. PMID- 9735634 TI - New model projects 11 percent fewer deaths in 20 years if U.S. smokers stopped smoking. PMID- 9735635 TI - HIV infections increasing among women and minorities. PMID- 9735636 TI - The role of the primary care physician in maximizing cognitive and behavioral recovery from moderate to severe pediatric traumatic brain injury. AB - Traumatic brain injury (TBI) is a major cause of death and the most common cause of acquired disability in children. Moderate to severe TBI typically results in cognitive deficits, and behavioral and psychosocial adjustment problems, sometimes compromising long term development. Although variable, considerable recovery can occur, especially over the first one to two years post-injury. Appropriate educational and psychological intervention is critical to positive outcome. The primary care physician (PCP) has a major role in monitoring progress and intervention, and therefore in maximizing outcome. The short term neurocognitive and neurobehavioral sequelae and recovery pattern for traumatic brain injury, and the role of the primary care physician in maximizing recovery based on risk factors are described. Some intervention resources are included. PMID- 9735637 TI - Pregnancy and the working woman: a review. AB - The potential impact of employment on pregnancy is an important issue that merits assessment, given the increasing numbers of women entering the labor force and continuing employment throughout pregnancy. A review of the literature evaluating the effects of employment on pregnancy is presented, including a brief historical and legal perspective, as well as employment statistics in the USA. There is emerging evidence from various studies suggesting that long working hours and prolonged standing may place a pregnancy at risk. Thus, physicians need to counsel their patients about these potential risks and provide recommendations for early modification of employment activities, in order to have a positive impact on pregnancy outcome. PMID- 9735639 TI - Monitoring the heart rhythm continuously for two years!: the implantable syncope monitor. PMID- 9735640 TI - Patients' bill of rights. PMID- 9735641 TI - Risks of cerebrovascular events related to open heart surgery. AB - Prevention of perioperative cerebrovascular injury in patients undergoing open heart surgery is a serious task for the surgeon, especially as age and severity of atherosclerotic disease increases. The most significant predisposing factors have been identified as existing carotid arterial disease or prior stroke, heavy calcification of the aorta, renal dysfunction, advanced age, and diabetes mellitus. We have studied a series of 600 open heart patients from 1992 to 1995 from the incidence of peri-operative stroke and mortality, evaluating 16 risk factors: heavy calcification of the ascending aorta, asymptomatic carotid disease, insulin-dependent diabetes mellitus, prior CVA, left ventricular function (ejection fraction of 20% or less), age greater than 70, renal dysfunction, transmural myocardial infarction, fluid balance index greater than 2500 ccs, smoking, type of procedure, emergency procedure, non-insulin-dependent diabetes mellitus, cardiopulmonary bypass time, gender, and hypertension Stroke occurred in 8 patients (1.3%), one of whom die postoperatively. Full or near-full recovery was experienced by 5 patients; 2 patients remained partially dysfunctional at the end of the study period. The operative mortality was 2.0% (12 patients); 10 deaths occurred in hospital and 2 following discharge within 30 days postoperatively. The risk of stroke was 15 times greater in patients over age 70; 16 times greater in older males (> or = 70 years); 5 times greater in patients with prior stroke or existing (asymptomatic) carotid artery disease; 8 times greater in patients with renal dysfunction; 4 times greater with a positive fluid balance index; and twice greater when cardiopulmonary bypass exceeded 110 minutes. Four of the stroke patients had diabetes mellitus. Two of 9 patients with heavy calcification of the aortic arch suffered cerebrovascular injury. Six or more of the risk factors studied were present in 81 patients; all 8 stroke patients (9.9%) came from this subgroup. The study suggests the importance of pre operative evaluation of cerebrovascular atherosclerotic disease and the minimal manipulation ("minimal touch" technique) of a calcific aortic arch. PMID- 9735642 TI - The Henderson Trust Lecture. The development of the vertebral canal and associated neuro-physiological abnormalities. AB - In this lecture I have attempted to demonstrate that the size of the lumbar vertebral canal has clinical importance. The canal develops very early in life, and impaired growth at this time affects other growing systems. The patient with spinal stenosis has more than a spinal disadvantage. Improved obstetric and childhood care has the potential not only to prevent some of the troublesome back problems, but also to influence the health and neurological status in adult life. I hope that the first Henderson Trustees would have been encouraged by this lecture. It supports some of the philosophy that stimulated an interest in Phrenology. In the lumbar spine at least, the container-the vertebral canal-seems to have an important relationship to the function of its neurological contents. PMID- 9735643 TI - The role of cell adhesion molecules in craniofacial development. AB - The last decade has seen classification and characterization of previously identified glycoproteins which have been associated with cell adhesion. More recently, it has been realized that the range of these molecules is even wider than originally thought. Cell adhesion molecules are important in early development and their role in craniofacial development is now apparent. Furthermore, the interaction of cell adhesion molecules in other developmental phenomena such as epithelial mesenchymal transformation (EMT), reinforces the suggestion that these molecules are important in the later stages of development, particularly organogenesis. This article reviews the role of cell adhesion molecules in embryogenesis, with a particular emphasis on foetal craniofacial development. PMID- 9735644 TI - Civilian abdominal gunshot wounds in Lagos. AB - This prospective study of 78 patients who sustained abdominal gunshot wounds was performed to evaluate the pattern of injuries, treatment outcome and the role of selective conservative management. Three (3.8%) patients died before laparotomy. Four (5.1%) patients with superficial wounds were managed by local wound care. Fourteen (18%) patients who had equivocal or minimal abdominal signs were selected for conservative management. Laparotomy was performed in 57 (73.1%) patients who presented with an acute abdomen. The commonly injured organs were the small bowel (56.1%), colon (38.6%), liver (22.8%) and stomach (19.3%). Prolonged injury to arrival and surgical intervention time were contributing factors to the high incidence of sepsis (63.2%) and mortality (22.8%) after laparotomy. Two patients selected for conservative management required delayed laparotomy, one of which was negative. A 10-fold increase in prevalence of abdominal gunshot wounds has occurred in our institution in the 1990s. Selective conservative management is feasible without the use of expensive investigations. PMID- 9735645 TI - Open versus closed diagnostic peritoneal lavage: a comparison on safety, rapidity, efficacy. AB - There is considerable debate between the proponents of open and closed diagnostic peritoneal lavage (DPL). A prospective study was undertaken on 130 patients submitted to DPL. We performed 55 (42.3%) closed and 75 (57.7%) open lavages with sensitivity and specificity of 100 and 96.6% for the former and 92.2 and 100% for the latter. The mean time for insertion of the catheter and initiation of fluid infusion was significantly less in the closed DPL group, and so were the number of cases with prolonged procedures. No intra-abdominal or wound complications were detected with either method, but there were 10 DPL failures due to inability to conclude the procedure successfully and derive a definite result. Eight of these (10.6%) belonged to the open group and two (3.6%) to the closed (P < 0.05). Our findings suggest closed DPL is as equally sensitive and specific as closed DPL, but is more expeditious and offers inconclusive results less often. Both procedures are useful and should be parts of surgical training. PMID- 9735646 TI - Surgical treatment of thyroid cancer: the Singapore General Hospital experience. AB - Patients with differentiated thyroid cancers generally have a good prognosis. This should be considered when deciding the extent of surgical resection. Radical surgery, however, may be appropriate in the control of locally advanced disease. An audit of 149 cases of thyroid cancer treated in the Department of General Surgery, Singapore General Hospital between October 1988 and June 1994 is presented. Particular attention is drawn to eight patients who underwent radical surgery. There were 111 (74.5%) women and 38 (25.5%) men. The median age was 45 years (range 12 to 83 years) and 80.5% of the cancers were papillary carcinomas, 14.8% follicular, 2.7% medullary and 2.0% anaplastic. Total or near-total thyroidectomy was the most common procedure for primary disease in about 75% of patients. Eight patients (5.4%) underwent radical surgery-four laryngectomy, one pharyngectomy and three median sternotomy-for tumour clearance. Morbidity included: wound complications in 2%; hypocalcaemia, transient in 16.8% and permanent in 3.4%; and hoarseness of voice in 8.1% with 4.7% having proved recurrent laryngeal nerve palsy. All three patients with anaplastic thyroid cancer died within 3 months. Of the eight who underwent radical surgery, three (37.5%) are alive and disease-free at median follow-up of 20 months. In a correctly selected group of patients with locally invasive differentiated thyroid cancer, aggressive surgery is appropriate, with acceptable morbidity and mortality. PMID- 9735647 TI - Appendicectomy and ulcerative colitis. AB - The aetiology of ulcerative colitis (UC) is unknown. However, much interest has been devoted recently to the relationship between appendicectomy and ulcerative colitis. A case-control study was conducted, comparing appendicectomy rates between 110 patients with UC (group 1) and 136 patients attending an orthopaedic clinic free from UC (group 2). The appendicectomy rates were 0.9% (group 1) and 10.3% (group 2), respectively (P < 0.002). The present study shows that patients with ulcerative colitis had rarely undergone appendicectomy before the first manifestation of colitis. Further research on this relationship is called for. PMID- 9735648 TI - Feasibility of pre-admission nurse clerking of patients with vascular disease. AB - A prospective study has been undertaken to determine the feasibility of nurse-led pre-admission clerking of patients with vascular disease. A total of 249 of 300 patients with planned admissions attended the clinic; 91% of patients with varicose veins, 83% of patients about to undergo endovascular procedures and 24% of patients awaiting arterial reconstruction were seen in the clinic. Patients with arterial disease were significantly more likely to rely on other people to bring them to the clinic than those with varicose veins. As a result of their age and frailty and their presenting symptoms, patients with arterial disease are less likely to benefit from a pre-admission clinic than patients with either varicose veins or general surgical disorders. PMID- 9735649 TI - Out-patient follow-up after routine surgery: a questionnaire study. AB - A questionnaire was sent to all consultant general surgeons and urologists in Wales to assess current practice in out-patient follow-up after surgery for nine commonly performed procedures. A further questionnaire was sent to a random sample of general practitioners in South Glamorgan to assess the possibility of GPs taking on responsibility for post-operative follow-up. This should, therefore, reduce the number of post-operative patients passing through the out patient department. Of the 58 (77%) consultants who responded, the percentage who routinely followed up patients with at least one post-operative visit was calculated. Of the 33 (66%) GPs who responded the percentage who were prepared to take responsibility for post-operative follow-up was calculated for patients having the same operations. There was agreement and disagreement between consultants and GPs, but not distinct pattern emerged. A larger study must be performed before the out-patient department is rationalized using protocols based on this study. PMID- 9735650 TI - The results of surgical techniques in hepatic hydatidosis: treatment with drainage versus treatment without drainage--a 6-year experience. AB - Fifty-nine consecutive patients who underwent surgery for hepatic hydatid cysts between 1 March 1988 and 31 July 1994, were included in this study. The aim was to compare the results of surgical techniques with respect to post-operative complications, morbidity and recurrence of the disease. Patients were divided into two groups. The first group (1) of patients (n = 30) were treated surgically without drainage and the second group (2) of patients (n = 29) were treated surgically with tube drainage of the cystic cavity. In the study, there were 12 (20.33%) male and 47 (79.66%) female patients, with an age range of between 16 and 85 years. In five of the patients a communication between the cyst and the biliary system was documented. 11.8% of the cysts were localized extrahepatically. Post-operative morbidity was 10% in group 1 and 44.7% in group 2 (P < 0.05). Average post-operative hospital stay was 8.5 days in group 1 and 18.6 days in group 2 (P < 0.05). During the follow-up period, which lasted from 1 to 6 years, recurrence rates were not significantly differentiated in the two groups (P > 0.05). PMID- 9735651 TI - The case for clinical audit of emergency readmissions after appendicectomy. AB - The aim of this study was to examine how the Scottish Office Clinical Outcome Indicator for emergency readmissions within 28 days of appendicectomy could be made more useful. Scottish Morbidity Record One (SMRI) data on all NHS discharges with a primary operative procedure of appendicectomy, and a linked file of all emergency readmissions within 28 days of discharge in 1993/94 were used in this study. It was found that 8783 appendicectomies were performed in 1993 and 1994, with 403 (4.6%) emergency readmissions within 28 days. A significantly higher proportion of emergency readmissions occurred where a 'normal' appendix was removed, and in interval, prophylactic and incidental appendicectomies. Age and length of initial hospital stay did not affect readmission rates. Hospitals performing more appendicectomies tended to have more readmissions [Spearman's rank correlation coefficient 0.639 (95% CI 0.430-0.782]. None of the five hospitals carrying out less than 17 appendicectomies in 2 years had an emergency readmission. Patients from more deprived areas were significantly more likely to be readmitted. The current performance indicator is useful in identifying patients readmitted to any hospital in Scotland, but needs to be supplemental with more clinical information. The numbers of appendicectomy patients would make this a feasible national clinical audit project, and the lessons learnt should be useful in promoting better practice for considerable numbers of patients of all ages. PMID- 9735652 TI - Modified Salter's innominate osteotomy. AB - Twenty paediatric patients with congenital dislocation of the hip were treated with a modification of the classic technique of Salter's osteotomy. Along with open reduction and femoral shortening osteotomy, the resected femoral segment, which measured 10-40 mm in width (mean 16 mm), was used to stabilize the Salter's osteotomy. The patients were aged between 2 and 9 years (mean 3.8 years). After a mean follow-up of 16 months (range 6-60 months) all patients had full bone healing and complete incorporation of femoral bone graft with the acetabulum. No patient had re-dislocation, subluxation or displacement of the graft. PMID- 9735653 TI - Screening times with image intensifier in orthopaedic trauma surgery. AB - Image intensifier technology has been of great benefit to orthopaedic, but not a benign aid. We have examined the image intensifier screening times for trainees and consultants over a seven month period. Consultants did not always have shorter screening times, and there were no differences between radiographers, Surgeons of all grades should depend more upon anatomical knowledge and spatial orientation, and less upon the image intensifier. PMID- 9735654 TI - Are abdominal radiographs still overutilized in the assessment of acute abdominal pain? A district general hospital audit. AB - Several studies have shown that plain film radiography (PFR) is unnecessary for most patients with abdominal pain. To evaluate the current-day utilization of PFR, we retrospectively reviewed 224 patients presenting to an emergency department with acute abdominal pain. Plain film radiography was performed in 55.8% (125/224) of patients, but only 10.4% (13/125) of these were diagnostic. Most patients with non-specific abdominal pain had radiographs (62%, 31/50), suggesting that PFR was being used as a routine investigation. Plain film radiography has little in the diagnosis of most causes of abdominal pain and should therefore not be used routinely. Confining radiography to patients with suspected gastrointestinal obstruction, perforation or ischaemia, unexplained peritonism, or renal colic would have included all our diagnostic films and reduced the utilization of PFR to 20.5%. The reasons for inappropriate requests and issues concerning the use of emergency radiography are discussed. Staff education, departmental protocols and increased out-of-hours ultrasonography facilities are recommended to reduce the inappropriate use of PFR. PMID- 9735655 TI - Surgical anatomy of the parathyroid glands in secondary hyperparathyroidism. AB - Parathyroidectomy is commonly required in patients with hyperparathyroidism secondary to end stage renal failure. Most descriptions of parathyroid anatomy are based on studies of normal glands. The details of parathyroid anatomy in 60 consecutive patients undergoing total parathyroidectomy for hyperparathyroidism secondary to renal failure are presented. Comparisons are made with the anatomical locations in series of normal glands. Based on these findings a strategy is proposed for the successful identification of all the parathyroid glands in patients undergoing parathyroidectomy for renal hyperparathyroidism. PMID- 9735656 TI - Anterior resection: right colon mobilization for colo-rectal anastomosis. AB - A variety of problems may compromise the safety of colo-rectal anastomosis when there is insufficient length of left colon available to provide well-vascularized bowel without tension. This may occur when there has been previous surgery or when the marginal artery is inadequate or when, following mobilization of the splenic flexure, there is insufficient length of bowel. This article is a description of a technique used to overcome such difficulties. PMID- 9735657 TI - On-table colonic lavage: an alternative. AB - A new technique of on-table colonic lavage is described. This technique has been used in those patients with inadequate bowel preparation at the time of elective or semi-elective surgery. The main difference in this technique compared with others previously described is that the lavage is performed after the resection and anastomosis of the colon is completed. Experience of this technique with four patients is described. PMID- 9735658 TI - Bacterial translocation in thioacetamide induced liver cirrhosis in rats. AB - Cirrhotic liver is predisposed to bacterial infections. Different species of bacteria including Escherichia coli, Enterobacter and Bacteroides fragilis were found to colonize thioacetamide-induced cirrhotic rat liver. Zinc treatment of the cirrhotic rats significantly corrected the histological and histochemical changes in the liver. However, this reversal with zinc treatment was not accompanied by any change in the bacterial colonies in the liver. The study shows that cirrhosis predisposes liver to bacterial colonization and the process is not reversible despite the partial reversal of the cirrhotic changes. PMID- 9735659 TI - Endovascular abdominal aortic aneurysm repair in the 'hostile abdomen'. PMID- 9735660 TI - Cell cycle regulators during human atrial development. AB - OBJECTIVES: The molecular mechanisms that regulate cardiomyocyte cell cycle and terminal differentiation in humans remain largely unknown. To determine which cyclins, cyclin dependent kinases (CDKs) and cyclin kinase inhibitors (CKIs) are important for cardiomyocyte proliferation, we have examined protein levels of cyclins, CDKs and CKIs during normal atrial development in humans. METHODS: Atrial tissues were obtained in the fetus from inevitable abortion and in the adult during surgery. Cyclin and CDK proteins were determined by Western blot analysis. CDK activities were determined by phosphorylation amount using specific substrate. RESULTS: Most cyclins and CDKs were high during the fetal period and their levels decreased at different rates during the adult period. While the protein levels of cyclin D1, cyclin D3, CDK4, CDK6 and CDK2 were still detectable in adult atria, the protein levels of cyclin E, cyclin A, cyclin B, cdc2 and PCNA were not detectable. Interestingly, p27KIP1 protein increased markedly in the adult period, while p21CIP1 protein in atria was detectable only in the fetal period. While the activities of CDK6, CDK2 and cdc2 decreased markedly, the activity of CDK4 did not change from the fetal period to the adult period. CONCLUSION: These findings indicate that marked reduction of protein levels and activities of cyclins and CDKs, and marked induction of p27KIP1 in atria, are associated with the withdrawal of cardiac cell cycle in adult humans. PMID- 9735661 TI - IgE production from the nasal polyp tissue: comparison between atopic and non atopic subjects. AB - OBJECTIVES AND METHODS: To confirm the local production of IgE antibody from the nasal polyp tissue, and to evaluate the difference between atopics and non atopics, nasal polyp tissues were taken from both 10 atopic and 10 non-atopic subjects. The tissue total IgE (tlgE) level was measured by enzyme-linked immunosorbent assay (ELISA) and serum tlgE level by radio-immunoassay. The tissue albumin level was measured by nephelometry, and serum albumin level by Bromocresol green method. RESULTS: The polyp tissue tlgE/albumin as well as serum tlgE/albumin ratio were significantly higher in atopics than in non-atopics (p < 0.05), with no difference in the albumin level between the two groups (p > 0.05). Three non-atopic subjects had high polyp tissue tlgE/albumin (> 10). A significant correlation was noted between serum tlgE/albumin and polyp tlgE/albumin (r = 0.46, p = 0.04). The ratio of polyp tlgE/albumin to serum tlgE/albumin was greater than 1 in all of the non-atopic subjects and 7 of 10 atopic subjects. CONCLUSION: These findings support the hypothesis that IgE antibody could be locally produced from the nasal polyp tissue of non-atopic subjects as well as atopic subjects. The possibility of an isolated local production of IgE antibody was suggested. PMID- 9735662 TI - Role of specific IgE, IgG and IgG4 antibodies to corn dust in exposed workers. AB - BACKGROUND AND METHODS: To evaluate the role of specific antibodies to corn dust (CD) and their relationship to respiratory dysfunction, we detected serum specific IgE(slgE) and IgG4(slgG4) antibodies by ELISA in 42 employees working in the animal feed industry and 27 unexposed controls. RESULTS: Our survey revealed that 15 (34.9%) subjects had work-related respiratory dysfunction associated with or without nasal symptoms. Among these subjects, eight had airway hyper responsiveness to methacholine. Significant differences were noted in slgE and slgG4 between exposed and unexposed groups (p = 0.04, p = 0.00 respectively), but no difference was noted in slgG (p = 0.1). Although there was no significant differences in the prevalence of specific IgE antibody between symptomatic (29%) and asymptomatic groups (19%, p = 0.55), the specific IgE levels were significantly higher in symptomatic workers than in asymptomatic workers (p = 0.03). Specific IgG antibody was detected in 1 (6%) symptomatic and 4 (15%) asymptomatic workers (p = 0.46). Specific IgG4 antibody was detected in 11 (73%) of symptomatic and 21 (78%) of asymptomatic workers (p = 0.90). The higher prevalence of slgG4 antibody was noted in workers with slgE antibody (p = 0.001). The correlation between slgG and exposure duration was significant (r = 0.36, p = 0.02). There was no association between the prevalence of slgE, slgG, and slgG4 to exposure intensity, smoking or atopic status. CONCLUSION: These results suggested that the existence of slgG and slgG4 might represent a response to CD exposure, and that some unexposed subjects had slgG to CD. Specific IgE might play a role in the development of respiratory symptoms. PMID- 9735663 TI - Detection of antibodies against DNA polymerase of hepatitis B virus in HBsAg positive sera using ELISA. AB - OBJECTIVES: DNA polymerase (pol) of Hepatitis B virus (HBV) includes 3 different domains such as terminal protein (TP), reverse transcriptase (RT) and RNase H. Humoral immune responses to each of these proteins have not been well documented previously, although antibody to pol was detected in serum of patients with chronic hepatitis B. We have constructed TP (amino acids 1-182), RT (amino acids 346-685) and RNase H (amino acids 690-832). METHODS: By ELISA using each protein expressed in E. coli as antigens, the corresponding antibodies were tested in serum from 40 patients with type B viral chronic liver diseases. (20 HBeAg positive and 20 HBeAg-negative). As negative controls, sera from 3 healthy young men were used. With the mean values of the OD, which were tested 4 times per each test sample and 3 times per each control sample, we considered to be positive if the mean OD of each test sample is 2-fold or higher than that of controls. RESULTS: Five of 40 sera (12.5%) contained one or two different antibodies detectable by this method: 4 of 20 HbeAg-positive sera (20%) and 1 of 20 HbeAg negative sera (5%). Anti-TP, anti-RT and anti-RNase H antibodies were detected in 2.5% (1/40), 10% (4/40) and 7.5% (3/40), respectively. Among 4/20 HbeAg-positive ELISA-positive sera, anti-TP, anti-RT and anti-RNase H were positive in 5% (1/20), 20% (4/20) and 10% (2/20), respectively, while 1 HBeAg-negative ELISA positive sera were positive only for anti-RNase H. CONCLUSIONS: These results suggest that the corresponding antibody responses to individual recombinant peptides derived from 3 domains of DNA polymerase may tend to be detected more frequently in HBeAg-positive sera than in HBeAg-negative sera from various patients with type B viral chronic liver diseases. PMID- 9735664 TI - Validity of the specialized columnar epithelium as a diagnostic criterion of the short segment Barrett's esophagus. AB - OBJECTIVE: In the areas where intestinal metaplasia of the stomach is highly prevalent, diagnosing Barrett's esophagus solely by the presence of specialized columnar epithelium in the distal esophagus may lead to many false positive diagnoses. The aim of this study was to test validity of the specialized columnar epithelium as a diagnostic criterion of the short segment Barrett's esophagus in Korea. METHODS: During routine gastroscopy, the length of columnar-lined esophagus was measured and biopsy samples were taken from the mucosa immediately distal to the squamocolumnar junction. Under light microscopy, alcian blue positive cells were identified. RESULTS: Prevalence of the specialized columnar epithelium in cases without the columnar-lined esophagus and with the short segment columnar-lined esophagus were 57.1% and 31.2%, respectively (P = 0.0281). The specialized columnar epithelium is frequently seen around the cardia in Koreans with or without the columnar-lined esophagus. CONCLUSION: Simple presence of the specialized columnar epithelium is not a valid criterion for a diagnosis of Barrett's esophagus. We propose that both the short segment Barrett's esophagus and the goblet cell metaplasia of the cardia might be grouped together under a title of "the specialized columnar epithelium around the gastroesophageal junction" as a potential preceding condition of adenocarcinoma around the cardia. PMID- 9735665 TI - Antigenic diversity and serotypes of Helicobacter pylori associated with peptic ulcer diseases. AB - OBJECTIVES: Clinical presentation of Helicobacter pylori (H. pylori) infection has marked variation mainly due to the strain diversity and host susceptibility. Although H. pylori is identified as a major risk factor for gastric and duodenal ulcers, the ulcerogenic or pathogenic strain has not been documented yet. The objective of this study was to investigate antigenic types of the ulcerogenic strain of H. pylori. METHODS: The sera of 64 patients were tested by Western blot using Helicoblot 2.0 for six major anti-H. pylori antibodies, together with CLO test and histological examination of gastric biopsy tissues. Thirty-five, nine and 20 patients had duodenal ulcer, gastric ulcer and chronic active gastritis, respectively. The antigenic types of H. pylori were analyzed in 54 patients with positive H. pylori infection. In this study, H. pylori was divided into four serotypes according to the presence and absence of CagA and VagA: type I; CagA (+) and VacA(+), type Ia: CagA (+) and VacA(-), type Ib: CagA(-) and VacA(+), and type II: CagA(-) and VacA(-). RESULTS: There was no difference in the number of bands for six antigens: 3.2 +/- 1.4, 3.0 +/- 1.2 and 3.1 +/- 1.4 in 35 duodenal ulcer, 7 gastric ulcer and 12 chronic gastritis, respectively. The band with 119 kDa was 90.7%, which was the most common band with the order of 35, 30, 26.5, 89 and 19.5 kDa. Type I, la and Ib were positive in 22.2, 42.6 and 27.8%, respectively, which were significantly higher than type II (p < 0.05). There was no difference in the positive rates of four urease subtypes between the four serotypes. PMID- 9735666 TI - p53 mutation in patients with ulcerative colitis in rectal biopsy. AB - OBJECTIVES: Long standing ulcerative colitis (UC) has been known to be one of the precancerous diseases of colorectal cancer. Although the frequent loss of p53 allele (LOH) and aneuploidy were reported as the molecular events in carcinoma and dysplasia known as the precursor of UC, p53 genetic alteration was not reported in indefinite dysplasia and UC involved mucosa in long standing UC. Therefore, we investigated the mutational inactivation of the p53 gene in UC patients who showed dysplastic mucosa, as well as non-dysplastic mucosa on H & E stain and, secondly, if there is p53 mutation, we examined the relationship between p53 alteration and clinical data. METHOD: Sixteen patients with UC who had different duration of colitis were studied by endoscopic examination with rectal mucosal biopsies, p53 gene alterations were detected by PCR-SSCP for exon 4-8 and immunohistochemical staining with p53 monoclonal antibody. RESULTS: Among 16 patients, 2 patients (12%) showed dysplasia on H-E stain. The p53 point mutations were detected in 4 (two dysplasia and 2 normal looking mucosa) on PCR SSCP. 4 patients who had p53 gene mutation were positive in immunohistochemical staining. With regard to clinical characteristics, these patients with p53 point mutation showed poor response to medical treatment. CONCLUSION: These results suggest that the p53 mutation may be an early molecular event of cancerous change in UC. PMID- 9735667 TI - Pheochromocytoma complicated with cardiomyopathy after delivery--a case report and literature review. AB - Pheochromocytoma in pregnancy is very rare but it is associated with very high maternal and fetal mortality. Therefore, it is important to include pheochromocytoma in the differential diagnosis of hypertension associated with pregnancy. It is difficult to make a diagnosis of pheochromocytoma in pregnancy before delivery. The characteristic symptoms of pheochromocytoma could be initiated during delivery because the process of delivery, general anesthesia, fetal movement, induce acute surge of catecholamine release, which could also induce cardiomyopathy. Early diagnosis and intensive care can affect the prognosis of cardiomyopathy induced by pheochromocytoma. Proper management with alpha-blockade, beta-blockade and angiotension converting enzyme inhibitor could acutely reverse the course of cardiomyopathy. PMID- 9735669 TI - Mercury inhalation poisoning and acute lung injury. AB - Acute mercury inhalation poisoning is a rare cause of acute lung injury. It is usually fatal because of progressive pulmonary failure. We experienced a patient with acute respiratory distress syndrome (ARDS) after illicit use of mercury vapor for hemorrhoid treatment; he developed acute chemical pneumonitis following exposure to mercury vapor. Prompt treatment with corticosteroids and penicillamine for acute chemical pneumonitis was instituted; radiologic pulmonary infiltrates disappeared within a week, but late phase neurologic sequelae and pulmonary interstitial fibrosis progressed. PMID- 9735668 TI - A case of arteriovenous type cardiac hemangioma. AB - Cardiac hemangiomas are rare primary tumors of the heart and constitute only 2.8% of primary cardiac tumors. They are classified into capillary, cavernous, epitheloid and arteriovenous type and the last one is the most uncommon type. We experienced a case of cardiac hemangioma which was diagnosed as arteriovenous type for the first time in Korea in the literature. The patient was a 54-year-old woman who presented with palpitation and anterior chest pain. The diagnosis was based upon coronary angiography which showed two tumor blushings located in the interatrial and interventricular septum with venous drainage to the coronary sinus and right atrium. Associated atrial fibrillation with rapid ventricular response was controlled with digitalis. PMID- 9735670 TI - A case of hemolytic uremic syndrome associated with Epstein-Barr virus infection. AB - The precise etiology of hemolytic uremic syndrome (HUS) is unknown. However, it has been associated with bacterial (Shigella, Salmonella, E. coli, S. pneumoniae), Bartonella, and viral (coxsackie, ECHO, influenza, varicella. Epstein-Barr) infections and with endotoxemia. Recently, we experienced a case of HUS in a 16-year-old boy who was in the acute phase of an Epstein-Barr virus (EBV) infection. He had typical manifestations of HUS and EBV infection. He also transiently presented disseminated intravascular coagulation. His renal dysfunction recovered by supportive care, including hemodialysis, plasmapheresis, antihypertensive medication and aspirin. We present this case with a review of the literature as the second report of HUS associated with EBV infection. PMID- 9735671 TI - A case of Behcet's disease with superior and inferior vena caval occlusion. AB - Behcet's disease is a chronic multisystemic disorder involving many organs and characterized by recurrent oral and genital ulcers and relapsing iritis. A case of BD with large vein thrombosis involving superior and inferior vena cava is presented. Large vein thrombosis in BD is not commonly developed and most commonly observed in the inferior or superior vena cava. A review of the literature emphasizes the rarity of the combined superior and inferior vena caval occlusion. Existence of extensive large vein occlusion in BD is associated with limited therapy and poor prognosis. PMID- 9735672 TI - A case of Marfan syndrome with acute monoblastic leukemia. AB - We report on an 18-year-old man who had both acute monoblastic leukemia and Marfan syndrome. A diagnosis of Marfan syndrome was established by those characteristics of arachnodactyly, ectopia lentis, mitral valve prolapse, and mitral regurgitation. Findings on bone marrow examination of the patient showed that most of nucleated cells were monoblasts and immunophenotype of those cells showed CD13+, CD33+, CD56+, and HLA-DR+. To our knowledge, this is the second report of leukemia in Marfan syndrome in the world. PMID- 9735673 TI - A case of CMV disease of the jejunum in a patient with non-Hodgkin's lymphoma. AB - CMV infection may occur anywhere in the gastrointestinal tract. Among the small intestine, ileum is the most common site of CMV disease and infection of jejunum is a rare one in patients with CMV gastroenteritis. Although rare, the reason why the recognition of this diagnosis is important is that it cause the lethal hemorrhage and perforation of gastrointestinal tract when its diagnosis and treatment was delayed. Rapid diagnosis are able to using the immunohistochemical stain in shell vial culture of infected specimen or peripheral neutrophils preparation in viremic patients within 8 to 36 hours. The treatment of choice is antiviral agent or surgical resection. We experienced a case of CMV disease of jejunum in patient with non-Hodgkin's lymphoma who showed severe ulceration in jejunum and massive intestinal hemorrhage, and he survived after successful treatment with segmental resection of jejunum and intravenous ganciclovir. PMID- 9735674 TI - Homocysteine Metabolism 2nd International Conference. Nijmegen, The Netherlands, 26-29 April 1998. Abstracts. PMID- 9735675 TI - The vitamin K-dependent proteins: an update. AB - Historically known for its role in blood coagulation, vitamin K also has been shown to be required for the physiologic activation of numerous proteins that are not involved in hemostasis. Over the last 20 years, vitamin K-dependent proteins have been isolated in bone, cartilage, kidney, atheromatous plaque, and numerous soft tissues. Although the precise mechanism of action of many of these proteins remains to be determined, their discovery has proven important from a physiologic point of view. PMID- 9735676 TI - Soy protein and cardiovascular disease: the impact of bioactive components in soy. AB - Soy protein, when substituted for animal protein in the diet, will lower blood cholesterol. Recent research also provides evidence that soy protein and/or isoflavones may improve endothelial functioning and attenuate events leading to both lesion and thrombus formation. PMID- 9735677 TI - Open or closed? A world of difference: a history of homocysteine research. AB - This article presents the research of the Nijmegen homocysteine team on birth defects and vascular disease. Hyperhomocysteinemia was found in women who gave birth to offspring with neural tube defects (NTDs) and other birth defects and in women with vascular disease. Elevated homocysteine levels in the blood plasma can be explained by lack of B vitamins (folic acid), mutation of the 5,10 methylenetetrahydrofolate reductase (MTHFR) genes, or both. Genetic mutations were found on the first chromosome (677 C T and 1298 A-C) and can explain up to 50% of the protective effect of folic acid against NTDs. The inborn error of methionine-homocysteine metabolism was also found in cases with recurrent early pregnancy loss, schisis, congenital heart defects, and vascular problems such as placental abruption, infarcts, and fetal growth retardation. One of the most exciting medical findings of recent years is that folic acid can prevent NTDs. This might also hold true for other birth defects and vascular disease. PMID- 9735678 TI - Comparable efficacy of hydrogenated versus nonhydrogenated plant sterol esters on circulating cholesterol levels in humans. AB - A recent study in The Netherlands compared the effects of margarine enriched with different vegetable oil sterols with margarine containing sitostanol-ester on plasma total, low-density lipoprotein (LDL), and high-density lipoprotein (HDL) cholesterol concentrations. Margarine with sterolesters from soybean oil (mainly esters from sitosterol, campesterol, and stigmasterol) was as effective as a margarine with sitostanol-ester in lowering blood total and LDL cholesterol levels without affecting HDL cholesterol levels. PMID- 9735679 TI - Assessing the changing diet of indigenous peoples. AB - Because a dietary transition is occurring among indigenous populations from traditional foods to more market (store-bought) foods, there are concerns about a rise in diet-related chronic disease. More research into dietary intakes of indigenous peoples is needed. When the use of longitudinal studies is not possible, the use of cross-sectional data to characterize the process of dietary change appears to be an appropriate way to assess change during rapid transition. PMID- 9735680 TI - Dietary fat, trans fatty acids, and risk of coronary heart disease. AB - Results from human feeding studies and recent large-scale epidemiologic surveys suggest that dietary trans fatty acids enhance the risk of developing coronary heart diseases. Despite a lack of accurate data regarding dietary intake of trans fatty acids, existing epidemiologic data and evidence from experimental feeding studies support the idea that lowering current intakes of trans fatty acids may lower the risk of coronary heart disease. PMID- 9735681 TI - The University Diabetes Outreach Project and the American Diabetes Association in collaboration with the Caribbean Food and Nutritional Institute 4th annual postgraduate course and international conference. Ocho Rios, Jamaica, March 5-8, 1998. PMID- 9735682 TI - S-O-S! When a malpractice insurer sinks, how do its doctors survive? PMID- 9735683 TI - Attitudes about cancer pain: a survey of 727 health care professionals in West Virginia. AB - To assess the attitudes of health care professionals in West Virginia about cancer pain, we utilized an 11-item questionnaire developed by DeWiessman and T.L. Dahl. We administered this questionnaire to 727 health care professionals and chi-square tests were used to assess correlations between attitudes on cancer pain and demographic characteristics (e.g. age, family history of cancer, sex, etc.). The majority of respondents believed that greater than 60% of cancer patients experience pain and that most patients were undermedicated with respect to pain. Age and having a family member with cancer were the major factors affecting attitudes. Respondents less than 46 years old compared to those 46 or older, were more likely to believe that most cancer patients were undermedicated; that addiction to narcotics is rare; and that the patient is the best judge of cancer pain intensity. Respondents with a family member with cancer were less likely to be concerned about addiction if a family member is given morphine. We conclude that educational efforts should be directed at altering attitudes regarding cancer pain in individuals age 46 years or older and those without a family history of cancer. PMID- 9735684 TI - Right ventricular endocarditis due to infection of pacemaker apparatus. AB - This article describes an unusual case of endocarditis in a 73-year-old man with a permanent pacemaker. Echocardiography revealed vegetations in the right ventricle below the tricuspid valve only, and he required open heart surgery for removal of the pacemaker system. This case demonstrates the potentially protracted course of infective endocarditis many years after pacemaker insertion. It also emphasizes the importance of echocardiography in diagnosing right ventricular vegetations; and the need for total replacement of the endocardial pacing system whenever it becomes infected. PMID- 9735685 TI - Obsessive-compulsive disorder following closed head injury. AB - Obsessive-compulsive disorder (OCD) is a rare sequelae of closed head injury. Frontal, occipital and fronto-temporal injuries are frequent among reported cases. Response to treatment is often poor. This article describes a patient with OCD and reviews relevant anatomic, neurochemical and psychologic aspects of patients with this condition. PMID- 9735686 TI - Acute low back pain findings and management in an academic medical center. AB - To compare medical records of patients treated for acute low back pain in the departments of Family Medicine, Internal Medicine, Occupational Medicine and Emergency Medicine in an academic medical center to determine if there was variation in patient population, diagnostic and treatment procedures and outcomes. Records were randomly reviewed using a standardized form for patients diagnosed with ICD 9 codes pertaining to back pain. Of the 96 patients with acute back pain seen in outpatient areas, 66 were seen by Family Medicine, 26 by Medical Group Practice (MGP), and four by Occupational Medicine. One hundred seven were seen in the Emergency Department. There was no significant difference in duration or type of pain or the type of findings or treatment. Very few had positive physical findings, (9% outpatient and 10% Emergency Department), but many more, (38% outpatient and 17% ED), had psychosocial findings (smoker, dissatisfaction with work, previous psychiatric history, psychosomatic history, or abnormal social adjustment) documented, Plain films of the lumbrosacral spine done in both practice settings did not change treatment. PMID- 9735687 TI - Insight on emphysema--the first 300 cases of surgical treatment. AB - Our experience with lung volume reduction surgery for emphysema now encompasses more than 300 cases, including several prospective trials. We have a 3.5% operative mortality rate and, with aggressive use of Heimlich valves over the past 6 months, an average hospital length of stay of 8 days. Proper patient selection is essential and can be based primarily on results of pulmonary function tests (PFTs), ventilation/perfusion (V/Q) scans, and computed tomography (CT) scans. We have found that bilateral is more effective than unilateral staple lung volume reduction surgery, which is in turn better than unilateral laser surgery. In patients with bilateral upper lobe disease, average FEV1 (forced expiratory volume in a 1-second interval) improvement is 82%; overall, it is 61% (range -33 to 217%). We conclude that lung volume reduction surgery can be performed safely with acceptable mortality and excellent clinical results in properly selected, motivated patients. PMID- 9735688 TI - Splenomegaly in 2,505 patients in a large university medical center from 1913 to 1995. 1913 to 1962: 2,056 patients. AB - Splenomegaly was studied retrospectively at the University of California, San Francisco, School of Medicine, on all patients (N = 2,056) for the years 1913 to 1962. The patients were classified into several diagnostic groups, and these groups were tested for statistical significance (chi(2)) with many clinical and laboratory variables to determine their predictive value. Hematologic disorders were associated with 57% of cases of splenomegaly and 81% of cases of massive splenomegaly. Among patients with splenomegaly, 19% had infectious diseases, 11% had hepatic diseases, and 9% had congestive or inflammatory disorders. The residual 4% were considered to have primary splenic disorders or a disorder of unknown cause. The commonest diseases associated with splenomegaly were hematologic (acute and chronic leukemias), infectious (malaria, endocarditis, and tuberculosis), hepatic (chronic liver disease), congestive (congestive heart failure), inflammatory (thyrotoxicosis), and other (cancers not metastatic to the spleen). The diseases most frequently associated with massive splenomegaly were the chronic leukemias. The disease with the highest incidence of massive splenomegaly was myelofibrosis (23 of 29 patients, 78%). Splenectomy was performed in 154 patients (7%), primarily for hematologic amelioration and hepatic hypersplenism. Hematologic diseases showed significant associations with lymphadenopathy, generalized lymphadenopathy, massive splenomegaly, and cytoses (P .001) and with progressive splenic enlargement (P < .02). Infectious diseases showed significant association with fever, and hepatic diseases showed significant association with abnormal results of liver function tests (P < .001). The findings of this retrospective study should be validated prospectively. PMID- 9735690 TI - Insulinoma--experience from 1950 to 1995. AB - Insulinomas are rare tumors that originate from the islet cells of the pancreas. The purpose of this study was to analyze our experience in patients with insulinoma and present our approach to these patients. Medical records of 67 patients treated at the University of California, San Francisco (UCSF) Medical Center, 56 surgically and 11 medically, from 1954 to 1995 were retrospectively reviewed. Presenting symptoms, physical findings, laboratory data, pre and intraoperative localization studies, operative management, operative success, and post-operative complications were analyzed. Among the entire cohort, there were 11 patients with Multiple Endocrine Neoplasia type I (MEN 1) and 7 patients with multiple tumors. 46 out of 48 patients (96%) having first operations for benign tumors and 5 out of 8 patients (63%) having reoperations for benign tumors were successful, as were 6 out of 12 patients (50%) having operations for islet cell carcinoma. Overall, preoperative localization studies were positive in only 46% of patients and therefore failed to improve our surgical outcome. Careful palpation with intraoperative ultrasonography gave the best localization results. Enucleation of solitary tumors is curative in sporadic cases and gives the lowest complication rate. In patients with MEN 1, subtotal pancreatectomy with enucleation of tumours from the pancreatic head and uncinate process is recommended over simple enucleation because of frequent multiple tumors. PMID- 9735689 TI - Splenomegaly in 2,505 patients at a large university medical center from 1913 to 1995. 1963 to 1995: 449 patients. AB - Splenomegaly was studied retrospectively at the University of California, San Francisco (UCSF), School of Medicine in 301 patients from 1963 to 1995 and compared with the UCSF service of the San Francisco General Hospital Medical Center (SFGH) in 148 patients from 1979 to 1994. The combined 449 patients were classified into several diagnostic groups and were studied by means of several clinical and laboratory associations. Hepatic disease in the percentage of patients at UCSF (with those at SFGH given in parentheses) was associated with splenomegaly in 29% (41%), hematologic disease, 32% (16%); infectious diseases, 16% (36%); congestive or inflammatory disease, 10% (4%); primary splenic disease, 6% (1%); other, 5% (1%); and cause unknown, 2% (1%). Massive splenomegaly occurred in 27% of the patients of the combined series, particularly in patients with hematologic diseases. The acquired immunodeficiency syndrome (AIDS) occurred in more than half of the patients with infectious diseases at SFGH and was four times frequent than in the patients at UCSF. The commonest diseases associated with splenomegaly were hematologic (lymphoma), hepatic (chronic liver disease), infectious diseases (AIDS and endocarditis), congestive (congestive heart failure), primary splenic (splenic vein thrombosis), and other (malignancy not metastatic to the spleen). In 11 patients with AIDS and massive splenomegaly, Mycobacterium avium complex occurred in 8 (73%). Splenectomy was performed in 117 patients (26%), primarily for hematologic amelioration. I conclude that for splenomegaly of unknown origin, the invasive procedure of choice for patients with hematologic associations may be a bone marrow biopsy; for hepatic association, a liver biopsy; and for infectious disease associations, a lymph node biopsy, before any consideration of a diagnostic splenectomy. PMID- 9735691 TI - Fundamentals--Rudolf Virchow and modern medicine. AB - The 19th century pathologist Rudolf Virchow was a physician, scientist, and revolutionary. The preeminent medical investigator of his day, Virchow remains best-known for his theory of cellular pathology, which laid the conceptual foundation for modern scientific medicine. Less appreciated are Virchow's numerous accomplishments in public health, anthropology, and European politics, including his quest for social justice and democracy in Imperial Germany. The study of Virchow's life and writings may provide contemporary physicians with a powerful role model as we grapple with the complexities of the modern medical enterprise. PMID- 9735692 TI - Best screening tests for prostate cancer. PMID- 9735693 TI - Urolithiasis. PMID- 9735694 TI - Epidemiology of prostate cancer. PMID- 9735695 TI - Sildenafil: a milestone in the treatment of impotence. PMID- 9735696 TI - Chronic lymphocytic leukemia in a patient with sickle cell anemia. PMID- 9735697 TI - Suppurative parotitis. PMID- 9735698 TI - Society and public health: crisis and rebirth. PMID- 9735699 TI - Isotachophoretic determination of bisoprolol, clonidine, disopyramide and tolazoline in human fluids. AB - Separation and determination of bisoprolol, clonidine, disopyramide and tolazoline in control serum and in human urine was investigated by capillary isotachophoresis. The drugs were separated by using the cationic electrolyte system. viz., sodium acetate buffer (pH 4.64) (c1 = 10 mM)-beta-alanine. The compounds were almost totally isolated from serum by solid-phase extraction using a Sep-Pak C18 cartridge. The recovery of compounds varied from 87 to 99%. The linear calibration range was studied to apply the method to real human fluids. The limit of determination of the drugs was 40.0 micrograms/ml serum. The limit of determination by direct sampling for bisoprolol is 3 micrograms/ml urine. PMID- 9735700 TI - Esters of cephalosporins. Part VI. Properties of the beta-form of 1-acetoxyethyl ester of cefuroxime. AB - Properties of a new crystalline beta-from of 1-acetoxyethyl ester of cefuroxime have been investigated and compared with known alpha-from and the amorphous form of this ester. Differences between these forms are discussed. PMID- 9735701 TI - Technology and biopharmaceutical availability of solid ocular inserts containing sulfadicramide and some promoters. AB - Technology for obtaining solid ocular inserts made of poly(vinyl alcohol), containing sulfadicramide and some sorption promoters, was worked out. The rate of drug release was studied by assuming the pseudo-first order kinetics to hold true. An isolated animal cornea was used to measure the coefficients of permeability and the efficiency of sulfadicramide penetration through these corneas. PMID- 9735702 TI - Synthesis and comparative anti-phlogistic potency of new proteinogenic amino acid conjugates of 2-[2,6-dichlorophenyl-1-amino]phenyl acetic acid "diclofenac". AB - New proteinogenic amino acids conjugates of 2-[2,6-dichlorophenyl-1-amino]phenyl acetic acid "Diclofenac", [I] were synthesized. Glycine methyl ester and L methionine ethyl ester were coupled with [I] via the active ester method to give the corresponding 2-[2,6-dichlorophenyl-1-amino]benzyl carboxy N-amino acid ester of the type [IIa, b], respectively, which were hydrolyzed in alkaline medium to yield the free amino acids [IIIa, b]. Condensation of IIIa with glycine methyl ester using a modified classical carbodiimide (DCCI) method gave the corresponding, "Diclofenac" glycylglycine methyl ester [IVa]. Hydrolysis of compounds IVa gives the corresponding acid Va. Thionation of compounds IIb and IVa by reaction with Lewesson's Reagent (LR), afforded the corresponding thio analogues (VIa and IVb). Interestingly, while retaining considerable comparative anti-phlogistic activity (anti-inflammatory and analgesic), the synthesized candidates proved to be practically nonulcerogenic in rats. PMID- 9735703 TI - Synthesis and anticonvulsant properties of new N-piperazinylalkyl imides of succinic acid. AB - A number of N-[(4-aryl)- or (4-methyl)-l-piperazinyl)alkyl]imides of 3-aryl or 3,3-pentamethylenesuccinic acid were synthesized and tested for anticonvulsant activity in the maximum electroshock seizure (MES) and pentylenetetrazole seizure threshold (scMet) tests. Structures of the novel compounds were confirmed by elemental and spectral analyses. PMID- 9735704 TI - Syntheses of some 4-[(4-chloro-5-methyl-2-methylthiophenyl)sulphonyl]-1 (aryl)semicarbazi des and N-[(4-chloro-5-methyl-2-methylthiophenyl)sulphonyl]-N' (4-chlorophenyl) urea with potential anticancer activity. AB - A series of 4-[(4-chloro-5-methyl-2-methylthiophenyl) sulphonyl]-1 (aryl)semicarbazides [IV-XI] and N-[(4-chloro-5-methylthiophenyl)sulphonyl]-N'-(4 chlorophenyl)urea [XII] were prepared and evaluated in vitro for anticancer activity [IV-VII, IX-XII]. All compounds exhibited weak [V-VII, IX] or moderate [IV, X-XII] activity against some human tumor cell lines, whereas the compound [XII] showed a relatively high activity (Table 3). PMID- 9735705 TI - Propranolol analog with the natural monoterpene structure as a potent antiarrhythmic drug. AB - Antiarrhythmic effects and intracellular electrophysiological properties of a new antiarrhythmic compound (-)trans-4-[2-hydroxy-3(N-isopropylamino)-propoxyimino] cis-car ane (9) were studied in several models of arrhythmia and in isolated guinea-pig myocardial preparations. Compound 9 prevented the aconitine-induced arrhythmia, reversed the ouabin-induced arrhythmia depressed the maximum rate depolarization (Vmax), and shortened the action potential duration (ADP) and the effective refractory period (ERP). The data indicate that compound 9 is an effective antiarrhythmic presumably with a class 1 B mechanism of action. PMID- 9735706 TI - Government nursing initiatives are not enough. PMID- 9735707 TI - Recognizing barriers to disabled people. PMID- 9735708 TI - Enteral feeding: an overview of indications and techniques. AB - Most patients requiring nutritional support can be successfully managed using enteral feeding, i.e. the provision of nutritional support via the gastrointestinal tract. Various routes of access are available and patients should be assessed on an individual basis by the multidisciplinary team to select the most appropriate method and decide on a suitable regimen. Careful management and regular monitoring of enteral feeding are required in order to ensure safe and cost-effective nutritional support; the use of standard protocols and guidelines is recommended. With appropriate training, monitoring and support, enteral feeding is best provided in the community for many patients, and the use of home enteral feeding is rapidly increasing. PMID- 9735709 TI - Rehabilitation: scope for improvement in current practice. AB - This article concludes this part of the rehabilitation series by looking at the nursing contribution to rehabilitation. It brings together the common elements which have emerged in the previous seven articles, identifying areas in which there is considerable scope for improvement to current practice. Indicative areas of knowledge and skill required for nurses to play a more active role are considered and a number of potential models are briefly described. PMID- 9735710 TI - A systematic approach to self-medication in older people. AB - This aim of this study was to evaluate the effectiveness of an individualized education programme on older patients' knowledge of prescribed medication. A questionnaire for assessment of medication knowledge was used to collect data from 15 patients before and after an individualized education programme which lasted approximately 2 weeks. Following the collection of pre-intervention data, an individualized education programme was developed for each patient. In addition to verbal instruction, each patient was provided with written information on his/her drug regimen. As part of the education programme each patient also had an opportunity to self-administer his/her medication. The results of the study showed that all participants scored higher in the post-test than the pre-test, although the degree of improvement varied between participants. The average increase in knowledge was 26%. Patient satisfaction with the education programme was also assessed using a questionnaire. Results suggested that there was a very high level of satisfaction with the programme. PMID- 9735711 TI - Pre-amputation assessment using Roy's Adaptation Model. AB - This article describes the use of Roy's Adaptation Model as a framework for the assessment of a 69-year-old man undergoing a right below-knee amputation. The model recognizes that individuals are subject to internal and external stressors which can lead to adaptive or ineffective responses. The practitioner can help the patient to make adaptive responses to these stimuli through nursing interventions. The components of the model are illustrated using the patient as a case study. A comprehensive assessment in the four adaptive modes (physiological, self-concept, role function and interdependence) is undertaken and nursing diagnoses are made. PMID- 9735712 TI - Do not resuscitate: an ethical dilemma for the decision-maker. AB - Since its introduction in the 1960s, cardiopulmonary resuscitation (CPR) has been universally available to all hospital patients unless the consultant in charge has specified a 'do not resuscitate' (DNR) order. The public perception of CPR has tended to be one of overoptimism, but this is not matched by the low survival to discharge ratio of approximately 1:10. In addition, there is the risk of prolonging suffering, compared with the quick and relatively painfree alternative offered by cardiac arrest. Decisions about resuscitation pose many ethical dilemmas for those involved and should take into consideration the patient's wishes, prognosis and quality of life. PMID- 9735713 TI - Tegagen alginate dressing for moderate to heavily exuding wounds. AB - Traditionally known as 'the mariner's cure', alginates have been used as wound dressings for many years. Tegagen (formerly Tegagel) is an alginate dressing with improved performance from 3M. It is designed for a variety of wounds with moderate to high levels of exudate. Alginates are a very cost-effective treatment option for flat and cavity wounds of this type. PMID- 9735714 TI - Clinical supervision for the development of nursing practice. AB - Nursing practice demands a range of problem-solving skills and abilities, and preregistration courses may only provide limited preparation. Clinicians need to draw on the skills and expertise of experienced colleagues in order most effectively to use their time and energies in a new role or clinical area. Experienced staff may enhance both their clinical expertise and their support of colleagues through the role of clinical supervisor. Clinical supervision has benefits for the supervisor in terms of professional development and for the supervisee as regards identifying and addressing clinical problems. According to the position statement on clinical supervision for nurses and health visitors published by the UKCC (UKCC, 1996) clinical supervision aims to identify solutions to problems, improve practice and increase understanding of professional issues. This article examines roles and responsibilities in clinical supervision and discusses the benefits for individuals and for health services. PMID- 9735715 TI - As entries into nursing fall questions must be asked. PMID- 9735716 TI - Children are not simply miniature adults. PMID- 9735717 TI - Factors influencing tissue viability nurse specialists in the UK: 2. AB - The literature review presented in the first part of this article (Vol 7(11): 648 57) revealed a lack of empirical data on specific contextual issues concerning the current practice of tissue viability clinical nurse specialists (CNSs) in the UK. This descriptive study utilizes focus groups to explore the experiences of tissue viability CNSs. It highlights the development of a facilitative style of expert practice within this group of nurse specialists and makes recommendations for successful role implementation. The findings suggest that specialist nurses are evolving a style of practice in congruence with the UKCC's vision of specialist practice (UKCC, 1994) and that these characteristics are likely to become more apparent over the next decade. PMID- 9735718 TI - Pressure area management in an orthopaedic setting. AB - Pressure area care for patients who have undergone a hip replacement is extremely difficult as patients' mobility tends to be restricted because of fear of dislocation. Orthopaedic surgeons influence the positional changes and techniques utilized by nurses in the postoperative period. This article reviews factors that prevent early rehabilitation and highlights methods of pressure relief for this patient group. Alternating-pressure mattresses are a main source of pressure relieving equipment. A pilot study using the Nimbus II mattress was carried out to establish the role of alternating-pressure mattresses in the postoperative management of patients undergoing elective/emergency hip replacement surgery. None of the patients in the pilot study suffered prosthetic dislocation and 87% did not develop pressure sores. Three patients did develop a grade 2 pressure sore postoperatively. The results reinforce the use of alternating-pressure mattresses in the postoperative management of patients undergoing hip replacement surgery. PMID- 9735719 TI - Providing paediatric palliative care: collaboration in practice. AB - One of the main aims of palliative care is to enable clients to receive and access services in a way that maximizes their choice in relation to where, when and how they receive care. To achieve this end, it is essential that statutory and voluntary care agencies collaborate to provide an effective range of services. This article offers for consideration the experience of a children's hospice service and a paediatric oncology outreach service who collaborated to provide a service for children requiring paediatric and terminal care. It identifies a number of elements which are important for positive and effective collaboration. PMID- 9735720 TI - Relative concerns associated with genetics and surrogacy. AB - Assisted reproductive technologies, incorporating artificial insemination and in vitro fertilization techniques, are widely practised across Europe. Concern about surrogacy prompted the UK Government to commission a review and consultation on surrogacy arrangements. The ensuing document was concerned primarily with the questions of payment to the surrogate, and the regulation of agencies involved in surrogacy arrangements. This author feels that more fundamental issues should first be debated by both health professionals and society. Genetic aspects of surrogacy merit special attention, particularly regarding the genetic contribution to parenthood and the ownership and use of genetic information. Health professionals need to be fully aware of the ethical implications of advances in genetics and technology when they are applied to assisted reproduction. PMID- 9735721 TI - Percutaneous endoscopic gastrostomy: psychological effects. AB - A mainly qualitative study, with some quantitative measurement tools, was undertaken to explore the psychological effects of percutaneous endoscopic gastrostomy (PEG) feeding on both patients and carers. A significant level of depression and stress was found in patients whose lifestyle had changed greatly; this was due partly to having to use a PEG and part to the underlying disease. Patients were, at the same time, grateful for the benefits to their nutritional state of having a PEG. A high level of stress was experienced by the relatives of patients whose functional and/or mental abilities had significantly changed for the worse and who had had a PEG tube placed. A need for more initial factual information and for ongoing practical and psychological support for both patients and carers was identified. PMID- 9735722 TI - The Clear Advantage urinary incontinence sheath for men. AB - All too often in the past urinary sheaths have been dismissed as having failed their user and carer simply because they have been poorly fitted or incorrectly measured. This article looks at the progressive development of the urinary incontinence sheath, from the various 'home-made' appliances that people with bladder problems conjured up, to today's more effective, more reliable products, such as the Clear Advantage all-silicone urinary incontinence sheath from Seton Continence Care. PMID- 9735723 TI - Helping communities to live more healthily. PMID- 9735724 TI - Stages of stroke: a model for stroke rehabilitation. AB - This article offers a model for stroke rehabilitation in both community and hospital settings which was developed as a result of observations made by staff working in both areas. It examines the effects of using more traditional models of stroke and neurological rehabilitation, where professional practice is more isolated, together with the effects of models for acquired brain injury that have identified positive outcomes. It was recognized by staff that a pragmatic model for stroke rehabilitation does not appear to be available. The Stages of Stroke model provides a 'map' with different routes to effective outcomes, helping those within stroke rehabilitation to recognize where they are and where they are going. It encourages a client-centred, not a service-centred programme. PMID- 9735725 TI - Aggression management. The pivotal role of nursing. PMID- 9735726 TI - Grandmothers who raise grandchildren. PMID- 9735728 TI - Quality of life and aging. PMID- 9735727 TI - Discharge planning for elderly patients. AB - The complex chronic health problems and functional limitations common in the elderly population place them at risk for complicated hospitalizations and discharge planning. The purpose of this study was to investigate the effectiveness of a discharge planning protocol in identifying elderly patients' home care needs. The sample in this quasiexperimental study consisted of 507 hospitalized patients age 65 years or older. The control group received the usual hospital discharge planning protocol. In the experimental group, nurse/social worker teams coordinated the discharge planning process, using an adapted form of the Discharge Planning Questionnaire (DPQ) to identify the home care needs of elderly patients. Thirty days after hospital discharge, both patient groups participated in a telephone survey to obtain information about health care problems they experienced during home recovery and their use of health care resources. The findings indicated that the majority of the elderly patients had functional dependencies, which required the help of another person to carry out daily household duties and provide assistance with basic needs, especially ambulation. These functionally dependent patients only received home care referrals about 50% of the time. These findings raise questions about current reimbursable services. Logistic regression analysis indicated that patients with increased functional dependency and patient problems during home recovery had a greater likelihood of rehospitalization and emergency department usage. This information about the home care of elderly patients after hospitalization supports the need for comprehensive functional assessment as part of discharge planning. This study also suggests that the nurse/social worker team can provide effective screening and discharge planning coordination of home care. Physician involvement and effective communication networks must be in place. PMID- 9735729 TI - Stress, social support, and health in frontier elders. PMID- 9735730 TI - Tuberculosis and the elderly. A community health perspective. AB - Tuberculosis is an ancient disease which has had a resurgence in the United States. Many elderly clients were exposed to tuberculosis as children and young adults and are still carrying the infection today. This article describes tuberculosis as it affects the elderly client in community and nursing home settings. The history, etiology, clinical manifestations, treatment, and prevention of tuberculosis infection and disease are presented. Because nurses can be very instrumental in controlling tuberculosis in the elderly, gerontological nursing implications are discussed. PMID- 9735731 TI - Testing the Uniform Needs Assessment Instrument for hospital discharge planning with older adults. AB - The Uniform Needs Assessment Instrument (UNAI) was developed to systematically assess the continuing care needs of high-risk older adults in response to the 1986 Omnibus Budget Reconciliation Act. Based on previous studies, a revised UNAI was tested with 103 hospitalized older adults, comparing usual discharge planning with the UNAI. High interrater reliability was obtained. The UNAI had high (> or = 85%) sensitivity and specificity when comparing needs identification on the UNAI with subjects' reported needs at 10 to 14 days after discharge. Overall, the UNAI was more effective (sensitive and specific). PMID- 9735732 TI - Reflex sympathetic dystrophy syndrome: an update. AB - RSDS is a complicated condition that requires early diagnosis and treatment for a favorable prognosis. The nurse should be alert to clinical signs in a patient with a traumatized area and report them to the physician as soon as possible. Management of the pain and establishment of trust will help promote successful physical therapy for the patient. The nurse should coordinate and manage the total care of the patient so that emotional and physical care can be at an optimum level for the patient and the patient's family. PMID- 9735733 TI - Nursing's role with amputee support groups. AB - An amputee support group is integral to holistic health care. A support group provides opportunities for peers and interdisciplinary team members to meet the unique needs of amputees. Nurses have an opportunity to be involved in the initiation and facilitation of an amputee support group within a health care agency or community. The purpose of this article is to discuss the role of nursing in developing an amputee support group. PMID- 9735734 TI - Stroke prevention screening program. AB - Stroke undoubtedly is one of the most devastating events experienced by humans. Stroke is the third leading cause of death in the adult population in the United States and the No. 1 cause of disability after 60 years of age. One out of four stroke victims die within 1 month. Many stroke victims would rather die than live without their dignity and quality of life. With the increase in the elderly population and the risk of stroke almost doubling every decade after 55 years of age, it is increasingly important to reduce the incidence of strokes. Stroke prevention screenings are an excellent opportunity to identify and educate persons who have significant stroke risk factors. Guidelines outlined by the National Stroke Association are described clearly in this step-by-step process of a stroke prevention screening. A unique modification of the National Stroke Association guidelines was the use of a carotid artery ultrasound screening to detect persons with significant carotid artery disease. This step in the screening replaced listening for carotid bruits. Seven carotid artery endarterectomies (of 385 participants) have been performed as a result of the screenings. Prevention of strokes through aggressive risk factor reduction cannot be overemphasized. PMID- 9735735 TI - Warm-Up Active Wound Therapy: a novel approach to the management of chronic venous stasis ulcers. AB - This preliminary study investigates the use of a radiant-heat bandage, Warm-Up Active Wound Therapy, as a new approach to the treatment of patients with chronic venous ulcers. Thirteen patients were randomly assigned to either conventional therapy or Warm-Up Active Wound Therapy for inpatient treatment of chronic venous stasis ulcers. Our results indicated that Warm-Up Active Wound Therapy is more effective than conventional therapy in healing chronic venous ulcers, and patients reported a significant reduction in wound pain. Warm-Up Active Wound Therapy also was found to be a safe treatment modality with no adverse events occurring in any patient. PMID- 9735736 TI - Port of call. PMID- 9735737 TI - Jobs for the girls. PMID- 9735738 TI - Are female nurses giving their male colleagues mixed messages? PMID- 9735739 TI - Panel games. PMID- 9735740 TI - What's in a name? Perhaps the safety of a child. PMID- 9735741 TI - The Daily Mail has been trying to raise public anxiety over teenagers, sex and the contraceptive pill to reckless levels. PMID- 9735742 TI - Australian nurses are no longer dominated by UK ideas, some of their problems are depressingly familiar. PMID- 9735743 TI - Voice-overs. AB - Hearing voices is distressing and frightening, but cognitive psychology can provide a range of interventions that enable people to live with their auditory hallucinations. Liam Clarke looks at ways to empower this patient group. PMID- 9735744 TI - Echoes of me. PMID- 9735745 TI - The invisible intruders. PMID- 9735746 TI - Floors to ceiling. Interview by Jo Carlowe. PMID- 9735747 TI - Pre-operative care--1. PMID- 9735748 TI - Hold your heads up. PMID- 9735749 TI - A battle to care. PMID- 9735750 TI - Choosing wound dressings. AB - This article continues our series looking at the knowledge needed for nurse prescribing. This week we discuss the use of wound dressings and take a brief look at the stages of wound healing. PMID- 9735751 TI - Cancer, chemotherapy and pregnancy. PMID- 9735752 TI - Community-based leg ulcer clinics: organisation and cost-effectiveness. AB - To establish the cost effectiveness of community leg ulcer clinics using four layer compression bandaging and the care provided by district nurses, a randomised controlled trial was set up in eight community-based research clinics in four trusts, funded by Trent Regional Health Authority. The main outcomes monitored were the amount of time needed to complete ulcer healing, patient health status, and recurrence. Satisfaction with care, use of services and personal costs were also monitored. PMID- 9735753 TI - A complement to care. PMID- 9735755 TI - The care and treatment of lung cancer. PMID- 9735754 TI - A personal experience of research. PMID- 9735756 TI - No time to waste. PMID- 9735757 TI - Learning to live with asthma. PMID- 9735759 TI - Expensive claim. PMID- 9735758 TI - A guide to asthma inhalers. PMID- 9735760 TI - Post haste. PMID- 9735761 TI - Stressed to death. AB - Nurse Richard Pocock took his own life after complaining of stress. Last week, legal history was made when the trust he worked for was ordered to pay compensation to his widow, writes David Payne. PMID- 9735762 TI - Women need to take folic acid sooner rather than later. PMID- 9735763 TI - Poor fare. PMID- 9735764 TI - Flour power. PMID- 9735765 TI - Defining what nursing is. PMID- 9735766 TI - Getting in on the act. PMID- 9735767 TI - The rise of little voice. PMID- 9735768 TI - All together now. PMID- 9735769 TI - In a league of their own. PMID- 9735770 TI - Don't accept the debrief. PMID- 9735771 TI - Pre-operative care--2. PMID- 9735772 TI - Nursing USA. PMID- 9735773 TI - The reporting of medical findings needs to be overhauled to prevent the media's obsession with health scare stories. PMID- 9735774 TI - The behavioural therapies. PMID- 9735775 TI - Rule bound. PMID- 9735776 TI - Smoking cessation programme for self-motivated patients. AB - Mr Mitchell felt that each weekly session had provided him with the impetus to continue for another week. He showed great determination to succeed and followed the recommended method, adapted to suit his own lifestyle. I felt professional satisfaction knowing that I had played a significant part in his transition from smoker to non-smoker. The method I used with Mr Mitchell was effective. However, it is only one of many methods available to smokers and health professionals. At present 22 patients have booked in for stop-smoking sessions. Six have completed the six sessions and of those, three remain non-smokers six months later. All three were self-referred and therefore highly motivated. Even one success is one more towards achieving the Health of the Nation target and is well worth the time and effort involved. PMID- 9735777 TI - Supporting clients with suicidal impulses in the community. AB - This article describes the development of community treatment team services and some of the methods they can employ in working with potentially suicidal clients in their own homes. PMID- 9735778 TI - Reporting of adverse drug reactions: practice in the UK. AB - As the range of available drugs becomes increasingly wide, it is ever more important for adverse drug reactions and interactions to be reported. With the extended role of the nurse moving closer towards nurse prescribing, this article seeks to demystify the adverse drug reactions reporting system in the UK, and explain the role of nurses. PMID- 9735779 TI - Easing children's pain. PMID- 9735780 TI - Night frights. PMID- 9735781 TI - Mixed images. PMID- 9735782 TI - Unfair division of labour. AB - Why is learning disabilities nursing still not seen as 'proper' nursing by so many? Ian Iles has had enough with this attitude and says it might be time for RNMHS to go it alone. PMID- 9735783 TI - Immunisation and vaccination for the under five-year-old. PMID- 9735784 TI - A dose of the UN's medicine. AB - Mariam Hamza hit the headlines when she was whisked to Britain from sanctions-hit Iraq to receive leukaemia treatment. Susannah Hall and David Olafimihan report from Iraq on the patients and nurses who were left behind. PMID- 9735785 TI - Young and vulnerable. AB - Nurses at Southampton University vaccinated 8,000 students after meningitis killed three students last year. Anne Gulland reports on what's being done to prevent another tragedy. PMID- 9735786 TI - On the record. PMID- 9735787 TI - Gray's elegy. Interview by Jane Salvage. PMID- 9735788 TI - Schizophrenia does have its plus side. PMID- 9735789 TI - On the other side of the research looking glass. PMID- 9735790 TI - Nowhere to go. PMID- 9735792 TI - Structural repairs. PMID- 9735791 TI - Who wants to be a millionaire? You don't. PMID- 9735793 TI - Snakes and ladders. PMID- 9735794 TI - My brilliant career. PMID- 9735795 TI - The agony and the ecstasy. AB - The issue of helping patients with their sexual needs has sparked heated debate. In the search for understanding, Hilary Francis describes how she assisted a disabled couple to have sex. PMID- 9735796 TI - Taking care of Kay. AB - A fortnight ago Peter Orr told how he coped with his wife's sudden brain haemorrhage. Here he describes how he enlisted help from the national media to win the care he believed was best for his beloved Kay. PMID- 9735797 TI - Catheter care--1. PMID- 9735798 TI - Thalassaemia. PMID- 9735799 TI - Face to face. Interview by Eileen Fursland. PMID- 9735800 TI - The immune system: hyperthyroidism. PMID- 9735801 TI - Patient records improve with unified case notes. AB - A study of multidisciplinary record-keeping at King George Hospital, Essex, resulted in an improvement in the quality of documentation and interdisciplinary communication. The system has now been implemented in all clinical areas. PMID- 9735802 TI - Effectiveness of child-health screening. AB - Screening for sensory problems in pre-school children often fails to detect a significant number of hearing problems sufficiently early. Evidence suggests neonatal screening is more effective and cost-effective than the infant distraction test at detecting congenital hearing loss. It is not yet clear how best to identify children with speech and language delay who will fail to progress without treatment. PMID- 9735803 TI - Nutrition and weight reduction. PMID- 9735804 TI - Health promotion for people with learning disabilities in the community. PMID- 9735805 TI - Tissue viability: the facts and the law. AB - Accountability remains an integral part of nursing, irrespective of occupational status or setting. The leaflet reminds nurses involved in direct care, as well as those in advisory, educational or promotional roles, of the need to consider and fulfil professional obligations towards a vulnerable and expectant public. The need to select resources that will support, not replace, skilled nursing is crucial. As nurses expand their practice they may face greater risk of litigation, but the boundaries should be clarified through job descriptions, employer approved policy statements and UKCC standards. Being asked to account for actions or omissions of care may be perceived as daunting. However, complaints may help drive quality improvement as well as staff development. A sense of perspective is needed to prevent defensive practice, allowing the potential of autonomous and accountable professionals being realised in the delivery of tissue viability services. PMID- 9735806 TI - Changing the records. AB - The aim of this study was to audit the documentation of pressure sore risk in a group of patients in a teaching hospital. This was done using a retrospective checklist assessment of patients in care of the elderly and orthopaedic wards. The study found there were delays in completing documentation on admission, that risk assessment was absent in 54% of patient documentation and that assessment of pressure areas was inadequate. PMID- 9735807 TI - Working the system. AB - This article describes the care given to a patients with a chronic leg ulcer during the author's supervised practice as a student district nurse. The Roper, Logan and Tierney (1996) model of nursing was used as a basis to assess, plan, implement and evaluate the nursing care given. PMID- 9735808 TI - Easing the symptoms. PMID- 9735809 TI - Tailor-made treatment. PMID- 9735810 TI - Film subjects win the day. PMID- 9735811 TI - Setting your career compass. PMID- 9735812 TI - The 1998 O/WM Buyers guide. PMID- 9735813 TI - GP payments for maternity care. PMID- 9735814 TI - High Court endorses women's right to refuse treatment. PMID- 9735815 TI - Compulsory caesareans: the last world? PMID- 9735816 TI - Hands off the breech. PMID- 9735817 TI - Antenatal HIV testing. Time for a change in practice. PMID- 9735818 TI - Is fat a pregnancy issue? PMID- 9735819 TI - Childbirth in women with a history of sexual abuse (II). A case history approach. PMID- 9735820 TI - Second stage. Midwives' views and practices. PMID- 9735821 TI - Campaign for food and drink in labour. PMID- 9735822 TI - The new Pregnancy Quitline. PMID- 9735823 TI - Birth positions in a large consultant unit. Encouraging trends. PMID- 9735824 TI - Who suffers when ignorance prevails? Stories about thalassaemia. PMID- 9735825 TI - HIV infection in pregnancy. PMID- 9735826 TI - Nobody home. PMID- 9735827 TI - Invest in breast together. PMID- 9735828 TI - Helping clients stop smoking: do brief interventions work? PMID- 9735829 TI - Childhood accidents: a major problem worth tackling. PMID- 9735830 TI - Attention deficit hyperactivity disorder. AB - In summary, ADHD is a mental disorder whose diagnosis is based on a child manifesting the symptoms of inattention, hyperactivity and impulsivity to the extent that the symptoms impair the child's ability to function. The main beneficial treatments are two nonspecific treatments, stimulant medication and behavioural interventions, preferably in combination. Dietary interventions have included diets that restrict allergenic foods, starting with a generally restricted diet and adding those foods that do not worsen the child's behaviour; a diet that restricts food additives and preservatives, referred to as the Feingold diet; and diets that restrict sugar. These dietary interventions either have not been proved to be efficacious or still require more study to determine their effects. PMID- 9735831 TI - Overcoming the pelvic flaw: exercises for continence. PMID- 9735832 TI - Common feeding problems in babies and children: 1. PMID- 9735833 TI - Hay fever: an update on current treatments. AB - Most hay fever sufferers can help themselves by taking steps to avoid high levels of pollen. Where eye symptoms are severe, sodium cromoglycate eye drops are recommended. Oral antihistamines are suitable for mild to moderate symptoms. For severe nasal symptoms, corticosteroid nasal sprays are useful. As with many medicines, use by expectant and breast-feeding mothers is best avoided. If symptoms are particularly troublesome, referral to a doctor is advisable. PMID- 9735834 TI - CPHVA Annual Professional Conference 1997. Pulling together for health: 2. PMID- 9735835 TI - [Vaccination: an experience from yesterday, today and, let us hope, tomorrow]. PMID- 9735836 TI - [In summer ticks are the thing]. PMID- 9735837 TI - [Drug distribution, still a current problem]. PMID- 9735838 TI - [Adaptation of the proper role and nursing diagnosis: what is the connection?]. PMID- 9735839 TI - [Awareness actions of nurses in the management of pain]. PMID- 9735840 TI - [The Mobile Emergency and Resuscitation Service: its role and efficacity]. PMID- 9735841 TI - [Criteria for the choice of a perfusion solution in emergency medicine]. PMID- 9735842 TI - [Subcutaneous administration of a therapeutic solution and/or a rehydration solution]. PMID- 9735843 TI - [Emergency materials for big or small infirmaries: selection of materials]. PMID- 9735844 TI - [The drowning patient: a patient unlike all others]. PMID- 9735845 TI - [Care of patients with lung grafts: lots of know-how]. PMID- 9735846 TI - [Nurses' aides in intensive care: a vast field of activities]. PMID- 9735847 TI - [Description of the functions of diploma nurses]. PMID- 9735848 TI - [Nurses in the land of football. Interview by Jean-Claude Gerez]. PMID- 9735849 TI - [I work at "La Beline"]. PMID- 9735850 TI - [Three cases of exceptional longevity]. PMID- 9735851 TI - [The centenarians. A model of successful aging]. PMID- 9735852 TI - [Emergencies. An episode of vomiting]. PMID- 9735853 TI - [The eyelids and their pathologies]. PMID- 9735854 TI - [AGGIR. Practical use. Geriatric Autonomy Group Resources Needs]. PMID- 9735855 TI - [The structural experience of a team]. PMID- 9735856 TI - [Night and patient planning in a geriatric institution]. PMID- 9735857 TI - [Help in getting up and going to bed]. PMID- 9735858 TI - [Urinary incontinence. Physiopathology of stress incontinence]. PMID- 9735859 TI - [The aged couple. How to adjust for the sick partner?]. PMID- 9735860 TI - Microparticle manipulation in millimetre scale ultrasonic standing wave chambers. AB - Ultrasonic standing wave chambers with acoustic pathlengths of 1.1 and 0.62 mm have been constructed. The chambers were driven at frequencies over the range 0.66-12.2 MHz. The behaviour of 2 microns diameter latex microparticles and 5 microns diameter yeast in the chambers has been elucidated. One (flow) chamber had a downstream laminar flow expansion section to facilitate observation of concentrated particle bands formed in the ultrasonic field. A second (microscopy) chamber allowed direct observation of band formation in the field and their characterisation by confocal scanning laser microscopy. Clear band formation occurs when the chamber pathlength is a multiple of half wavelengths at the driving frequency, so that the chamber rather than the transducer resonance has the most influence on band formation in this system. Band formation occurred in half-wavelength steps from a position one quarter of a wavelength off the transducer to a band at a similar distance from the reflector. Ordered band formation was preserved by the laminar flow in the expansion chamber, although bands that formed very close to the wall were dissipated downstream. The microscopy chamber provided evidence of significant lateral particle concentration within bands in the pressure nodal planes. The approaches described will be applicable to the manipulation of smaller particles in narrower chambers at higher ultrasonic frequencies. PMID- 9735861 TI - Sonoporation of erythrocytes by lithotripter shockwaves in vitro. AB - Sonoporation of red blood cells was examined in relation to cavitation-induced hemolysis. FITC-dextran at 580,000 MW was added to suspensions of canine erythrocytes and the mixture was exposed to lithotripter shockwaves. Exposure at 5% or 50% hematocrit in PBS or 50% in plasma yielded not only hemolysis but also FITC-dextran uptake in surviving cells. Hemolysis increased with increasing numbers of shockwaves. The numbers of cells with fluorescent dextran uptake remained roughly constant for 250-1000 shockwaves, but this represented an increasing percentage of the surviving cells. In addition, fluorescent microspheres formed spontaneously in samples with hemolysis. An air bubble was needed in the chamber to obtain substantial effects, implicating the cavitation mechanism. The exposure-response trends could be modeled by simple theory for random interaction of the cells with bubbles. PMID- 9735862 TI - Selective susceptibility of CD34-expressing cells to acquire flow cytometric features of apoptosis/necrosis on exposure to an ammonium chloride-based red blood cell lysing reagent. PMID- 9735863 TI - Diphtheria toxin A gene-mediated HIV-1 protection of cord blood-derived T cells in the SCID-hu mouse model. AB - The reconstitutive potential of CD34+-derived cord blood (CB) cells, transduced with a regulated diphtheria toxin A (DT-A) chain gene, was examined in SCID-hu mice harboring a conjoint organ composed of human thymus and liver (thy/liv). The DT-A-transduced cells, injected directly into the thy/liv organ, showed the same engraftment potential as control CB cells transduced with the non-DT-A parental vector. CB cells, distinguishable from the thy/liv cells by the HLA marker B7, were preferentially maintained in ex vivo culture. In the thy/liv organ, the engrafted CB cells represented >80% of the total cells. A majority of cells (>70%) in the thy/liv organ were also CD4+CD8+, as would be expected of maturing thymocytes. The incidence of double-positive cells was highest at 44 days (compared with 30 days and 80 days) after injection of CB cells. This suggested that a minimum time was required to achieve optimal proliferation of cells in the thy/liv organ but that, at later times, all of the early cells had matured. Thus, the population used for engraftment contained early cells but not self-renewing cells. The double-positive cells matured rapidly into single-positive cells (either CD4+ or CD8+) when placed in ex vivo culture. Marked cells (neo+) could readily be detected in the thy/liv-derived cells. The cells transduced with DT-A showed long-term protection in ex vivo culture against HIV T lymphotropic isolate NL4-3. This study shows that DT-A-transduced cells had no apparent disadvantage in engraftment of the thy/liv organ and did not have any toxic effects in vivo. Such cells were protected against HIV infection even when challenged more than 2 months after transduction and after a 44-day engraftment period in the thy/liv mice. These data support the feasibility of toxin gene therapy as a strategy for HIV infection. PMID- 9735864 TI - Spontaneous silencing of humanized green fluorescent protein (hGFP) gene expression from a retroviral vector by DNA methylation. AB - We have constructed a functional murine leukemia virus (MLV)-derived retroviral vector transducing two genes encoding the autofluorescent humanized green fluorescent protein (hGFP) and neomycin phosphotransferase (Neo). This was done to determine whether hGFP could function as a marker gene in a retroviral vector and to investigate the expression of genes in a retroviral vector. Surprisingly, clonal vector packaging cell lines showed variable levels of hGFP expression, and expression was detected in as few as 49% of the cells in a clonally derived culture. This indicated that hGFP expression was silenced in individual cells. This silencing could be diminished by selective culturing of the vector packaging cells with the neomycin analog G418 and was reduced by a 3-day treatment with the demethylating agent 5-azacytidine. The 5-azacytidine effect was transient, and hGFP expression in the vector packaging cells returned to untreated control levels within 2 weeks. Using flow cytometric analysis, hGFP expression was detected in up to 15% of transduced MT4 cells (a CD4+ lymphocytic cell line) after coculturing with packaging cells for 4 days. A 3-day postcoculture treatment with 5-azacytidine was shown to increase the hGFP-expressing MT4 cells from either 10.4% to 11.6% or 3.7% to 4.8%, corresponding to an increase in observed transduction efficiencies of 12% and 30%, respectively. These results indicate that silencing of gene expression from a retroviral vector may result from DNA methylation and occurs rapidly after transduction. PMID- 9735865 TI - Concurrent partial body radiation prevents cytokine mobilization of blood progenitor cells: an effect mediated by a circulating factor. AB - Mobilization of stem and progenitor cells into blood, which facilitates the collection of blood-derived autograft and allograft products, can be accomplished with administration of myelosuppressive chemotherapy, hematopoietic growth factors, or both. Autologous donor indifference to mobilization attempts has been correlated with prior administration of chemotherapy and radiation therapy. To investigate whether concurrent administration of radiation therapy inhibits mobilization, five daily injections of a potent combination of mobilizing cytokines, 500 U/kg erythropoietin (EPO) plus 15 microg/kg G-CSF, were administered each morning to Balb/c mice. Each afternoon, a 2 Gy fraction of Co 60 radiation was administered to either the lower limb or the upper or lower hemibody. Each day, mice were necropsied, and blood stem cell mobilization was determined by assaying the number of hematopoietic colony-forming cells in the blood and in the spleen. Unirradiated cytokine-injected mice showed a significant mobilization effect evident as increased colony-forming cells in blood and spleen compared with saline-injected unirradiated controls. The irradiated mice showed markedly inhibited or absent mobilization regardless of the part of the body irradiated. To investigate the mechanism of radiation-induced mobilization inhibition, heparinized plasma was obtained from mice whose lower bodies were irradiated with 2 Gy 18 h previously, and 0.5 ml was injected i.v. into intact mice 10 min before they received 15 microg/kg G-CSF and 500 U/kg EPO. Unlike mice that received G-CSF + EPO only and showed mobilization of progenitors from marrow to spleen, recipients of plasma from irradiated mice before and after cytokine administration showed significantly reduced mobilization of progenitors. Thus, radiation-induced inhibition of stem cell mobilization is mediated by an unidentified circulating factor. PMID- 9735866 TI - Use of collagen for standardization of PBSC graft quality evaluation: a multicenter comparative analysis of commercial collagen-based and methylcellulose based colony-forming unit (CFU) assay kits. AB - The colony-forming unit-granulocyte-macrophage (CFU-GM) assay, an essential test in evaluation of the quality of autologous grafts of hematopoietic stem cells, has yet to be standardized. With this aim in view, we carried out a multicenter study of five commercially available culture kits for CFU-GM evaluation. Four kits were methylcellulose-based (H4431, H4434, H4435, StemBio1d) and one was collagen-based (EasyClone-Multi). Using fresh and frozen samples of PBSC grafts, we compared CFU-GM and burst-forming unit-erythrocytes (BFU-E) growth using the EasyClone kit to each of the methylcellulose kits. BFU-E and CFU-GM clonogenicity of both fresh and frozen PBSC was clearly inferior with the H4431 kit, which provides conditioned medium only. CFU-GM numbers obtained with fresh and frozen PBSC samples were significantly higher with the EasyClone kit than with the H4434 and StemBio kits. BFU-E numbers were also higher with the EasyClone kit, but only when colonies were scored after May-Grunwald-Giemsa (MGG) staining. Finally, although the H4435 kit provides higher doses of recombinant cytokines than the EasyClone kit, CFU-GM and BFU-E numbers obtained for fresh or frozen PBSC with both kits were similar. In addition, CFU-GM and BFU-E numbers correlated well with CD34+ cell numbers for all five kits for both fresh and frozen PBSC. In summary, our study shows that the EasyClone-Multi and H4435 kits provide the best CFU-GM growth. The collagen-based EasyClone kit has the additional advantage of allowing gel staining and storage, which facilitates colony identification and, more importantly, makes gel exchange possible for standardization of the CFU-GM assay. PMID- 9735867 TI - High-sensitivity immunocytologic analysis of neuroblastoma cells in paired blood and marrow samples. AB - High-sensitivity immunocytochemistry was used to evaluate the relative frequency of neuroblastoma cells in bone marrow and peripheral blood in patients with neuroblastoma (NB). A total of 51 concomitant paired blood and marrow samples (102 total) from 35 patients with NB (age 4 months-31 years; stage 29 stage IV, 4 stage III, 2 stage IVS; 14 at diagnosis, 18 in relapse, 12 during treatment, and 7 off-therapy) were analyzed. Cytospins containing up to 10(6) cells each were prepared using the mononuclear cell (MNC) fraction. For immunocytologic staining, a primary mouse monoclonal anti-GD2 antibody (3F8), a secondary antimouse biotinylated antibody, and a streptavidin-alkaline phosphatase complex were used. A minimum of two cytospins containing a mean of 1.4 x 10(6) total MNCs was analyzed in addition to a negative and a positive control. No circulating tumor cells were detected when the concomitant marrow samples were negative or had <10 positive cells per 106 MNC (23 of 51 samples). Of the 18 marrow samples positive at 10-10,000 cells per 106 MNC, 6 had detectable NB cells in the corresponding blood sample, whereas for marrow samples with >10,000 NB cells per 10(6) MNC (1%), the concomitant blood sample was positive for 9 of the 10. When both marrow and blood samples were positive (15 BM-PB pairs), NB cell frequency was significantly lower in blood, with a mean difference of 2.14 logs (median 2.22, range -0.16-4.8, standard error 0.38). In patients with NB, circulating tumor cell frequencies seem to be substantially lower than in concomitant marrow samples, with a mean difference of >2 logs. PMID- 9735868 TI - Analysis of engraftment kinetics in pediatric patients undergoing autologous PBPC transplantation. AB - We sought to analyze factors that affect the engraftment kinetics following autotransplantation with PBPC mobilized by filgrastim (G-CSF). Forty-six consecutive pediatric patients with hematologic malignancies (n = 23) or solid tumors (n = 23) underwent autologous PBPC transplantation after myeloablative therapy. PBPC were mobilized using G-CSF alone. All patients received G-CSF after PBPC infusion. Factors potentially influencing the neutrophil and platelet engraftment were examined using univariate and multivariate analysis. All patients experienced rapid hematopoietic recovery, with a median of 9 days (range 7-15) to achieve a neutrophil count of 0.5 x 10(9)/L and a median of 15 days (range 9-37) to achieve a platelet count of 20 x 10(9)/L. The most important predictive factor of both platelet (p = 0.002) and neutrophil (p = 0.0001) recovery was the number of CD34+ cells infused. Patients receiving > or =5 x 10(6)/kg CD34+ cells had a more rapid hematopoietic recovery (p < 0.001) than those receiving a lower cell dose. The CD34+ cell dose is the most important predictive factor for engraftment kinetics after PBPC transplantation. Although a minimal CD34+ cell dose could not be defined, a dose > or =5 x 10(6)/kg CD34+ cells may be optimal to ensure rapid neutrophil and platelet recovery. PMID- 9735869 TI - A comparison of two different systems for CD34+ selection of autologous or allogeneic PBSC collections. AB - This study compared CD34 selection procedures using the CellPro CEPRATE and the Baxter Isolex 300 systems. Thirty-two procedures were performed, 19 CEPRATE and 13 Isolex. Median starting CD34 percentages were (CEPRATE/Isolex) 0.80% (range 0.24%-7.73%) and 0.85% (range 0.27%-10.17%), respectively (p = 0.788). After selection, there was a highly significant difference in purity of the product (CEPRATE/Isolex), 54% and 82% respectively (p < 0.0001). There was no significant difference in median recovery (CEPRATE/Isolex), 43% and 50%, respectively (p = 0.383). The starting CD34 percentage influenced the purity of the final product, and at high and low starting percentages, the Isolex produced superior purity. Improved efficacy of T cell depletion was observed with the Isolex, a median log depletion of 3.4 compared with 2.9 for the CEPRATE system (p = 0.012). In conclusion, the Isolex 300i produced a significantly higher purity CD34+ fraction, even at starting CD34+ levels of <0.5%, with no significant difference in recovery when compared with the CEPRATE system. The associated log T cell depletion is significantly improved with the Isolex system, with possible implications for use in CD34-selected allogeneic transplants. PMID- 9735870 TI - Effects of long-term storage at -90 degrees C of bone marrow and PBPC on cell recovery, viability, and clonogenic potential. AB - Autologous BM and PB HPC are usually stored from weeks to months until reinfusion after myeloablative chemotherapy. HPC have been stored for up to 16 months at -90 degrees C, using a mixture of 5% DMSO, 6% hydroxyethyl starch (HES), and 4% HSA as a cryoprotectant. Long-term storage (LTS) has usually entailed rate-controlled freezing using 10% DMSO and preservation in liquid nitrogen. The effects of LTS at -90 degrees C on the in vitro cell recovery, viability, and colony-forming unit-granulocyte macrophage (CFU-GM) clonogenic potential of autologous HPC that were not transplanted was studied. Sixteen BM and sixteen PB HPC had been cryopreserved for a median of 53 months (range 27-71) and 35 months (range 26 78), respectively. Samples of frozen HPC were thawed after 48 h, and the nucleated cell count, viability by trypan blue exclusion, and culture for CFU-GM were obtained. Following LTS, the cells were thawed and examined using the same assays. No difference in the median percentage recovery of nucleated cells was found in either the BM or PB HPC between the samples stored for 48 h and after LTS (5.73 x 10(9) versus 5.61 x 10(9) and 6.20 x 10(9) versus 5.78 x 10(9), respectively). In addition, no difference in median percentage viability was found in either the BM or PB HPC sampled at 48 h and at the end of LTS (75% versus 74% and 75% versus 76%, respectively). Finally, the median number of CFU GM cultured from BM HPC at 48 h was 2.41 x 10(5) (range 0.33-11.01 x 10(5)) and at the end of LTS was 1.93 x 10(5) (range 0.32-10.55), representing a median recovery of 93% (range 19%-308%). Similarly, the median number of CFU-GM cultured from PB HPC was 1.66 x 10(5) (range 0-50.57) and at the end of LTS was 0.93 x 10(5) (range 0-44.9), representing a median recovery of 80% (range 36%-165%). This difference in percentage recovery was not significant (p = 0.514). There was poor correlation between the number of nucleated cells harvested and the percentage recovery of nucleated cells, cell viability, or CFU-GM for either the BM or PB HPC. Similarly, there was poor correlation between the number of CFU-GM in the harvest and their percentage recovery following LTS for both BM and PB HPC. Finally, there was poor correlation between the storage time of the BM or PB HPC and the percentage recovery of nucleated cells, cell viability, and CFU-GM. These data suggest that LTS of HPC at -90 degrees C is not associated with decreased recovery of nucleated cells or in vitro viability and is associated with only a modest decrease in clonogenic potential. This indicates that storage of HPC at -90 degrees C for periods in excess of 3 years is possible. PMID- 9735871 TI - Camptothecin and taxol: discovery to clinic. AB - Camptothecin (CPT) is a pentacyclic alkaloid isolated from wood and bark of Camptotheca acuminata. Initially it was found to be highly active in a number of mouse in vivo cancer assays. Subsequently, CPT was found to uniquely inhibit an enzyme, topoisomerase I, which is involved in DNA replication. A number of CPT analogs are in advanced clinical trial, and two, Topotecan and CPT-11, have been approved for marketing by the FDA. taxol, a taxane alkaloid, was isolated from Taxus brevifolia. Taxol is a highly cytotoxic compound active in several mouse antitumor assays. It was subsequently found to uniquely inhibit tubulin, a protein involved in mitosis. After clinical evaluation, it has become the drug of choice for treatment of ovarian cancer. PMID- 9735872 TI - Paclitaxel (Taxol): a success story with valuable lessons for natural product drug discovery and development. AB - The discovery and development of paclitaxel, which covered a time span of some 30 years, has provided some important lessons for those involved in natural product drug discovery and development. These include the adoption of novel screens as they become available, the elucidation of mechanisms of action, and addressing the supply issue at an early stage of development. These issues, as applied to paclitaxel, are illustrated. The development of the NCI human cancer cell line screen, and its application to mechanistic studies through use of COMPARE analyses, are discussed, as is the production of the marine-derived anticancer agent, bryostatin 1, which provides another illustration of a successful approach to solving a supply issue. The history of the development of paclitaxel also illustrates the importance of multidisciplinary collaboration, and the various mechanisms used by the NCI Developmental Therapeutics Program for promoting such collaboration are presented. PMID- 9735873 TI - Prospecting for potentially new pharmaceuticals from natural sources. AB - Many new natural product-derived pharmaceutically active compounds and compositions, each effective in treating an array of diseases and maladies including various tumors and HIV, have been reportedly isolated from different sources of vegetation, including the bark of yew trees, needles, leaves, fungi, and cell culture of many different species; vegetables such as West African yams; and Chinese and Indian herbs. Other sources include vegetation from South American rainforests. Many of the sources of such natural products are historical in nature and/or are known from folklore. Recent studies have provided potentially new biodiverse pharmaceutical compounds such as paclitaxel, which is obtainable from several species, including various portions of T. brevifolia, the Western yew tree, and other yew species such as T. baccata, T. cuspidata, T. wallichiana, T. media, T. canadensis, T. chinensis and T. yunnanensis as well as from T. wardii, T. capitata, T. brownii, T. gem, T. globosa, T. floridana, T. hicksii, T. densiformis, and T. darkgreen spreader, in addition to cultured plant cells and fungi. The novel compounds and their semisynthetic brominated and chlorinated analogs prepared from T. yunnanensis extract show strong activity against several types of tumors. PMID- 9735874 TI - Noncariogenic intense natural sweeteners. AB - There is a definite relationship between the dietary consumption of sucrose and the incidence of dental caries. Noncaloric sucrose substitutes for use in the sweetening of foods, beverages, and medicines may be either synthetic compounds or natural products. In the United States, four potently sweet artificial sweeteners are approved, namely, saccharin, aspartame, acesulfame potassium, and sucralose. Highly sweet plant constituents are used in Japan and some other countries, including the diterpene glycoside stevioside and the protein thaumatin. Recent progress in a research project oriented towards the discovery and evaluation of novel potentially noncariogenic sweeteners from plants has focused on substances in the sesquiterpenoid, diterpenoid, triterpenoid, steroidal saponin, and proanthocyanidin structural classes. The feasibility of using Mongolian gerbil electrophysiological and behavioral assays to monitor the sweetness of plant extracts, chromatographic fractions, and pure isolates has been investigated. An in vivo cariogenicity study on the commercially available natural sweeteners stevioside and rebaudioside A has been carried out. PMID- 9735875 TI - Report from the Biological Stain Commission: FDA issues final rule for classification/reclassification of immunochemistry (IHC) reagents and kits. PMID- 9735876 TI - Staining paraffin extracted, alcohol rinsed and air dried plant tissue with an aqueous mixture of three dyes. AB - A staining solution containing alcian blue 8GX, Bismarck brown Y and safranin O was prepared with 0.1 M sodium acetate buffer, pH 5.0. Paraffin was extracted with MicroClear solvent from 10 microm tissue sections mounted on slides. Paraffin solvent was removed by rinsing with isopropanol, and tissues were air dried. Slides with bare dry tissue sections were immersed in the triple stain and structures could be distinguished within 30 min as follows: nonlignified cell walls, blue; lignified cell walls, nuclei and chloroplasts, red; and cuticle, brown or yellow-brown. Excess staining solution was removed by rinsing with tap water, and the tissues were air dried again. Coverslips were affixed with resin over the stained dry tissues. This novel procedure was tested with immature tomato fruit, mature apple fruit, and various leaf and stem specimens of dogwood, laurel, pawpaw, poinsettia and zonal geranium. PMID- 9735877 TI - Chimeric antibodies with specificity for tumor antigens: demonstration of in situ localization to tumors after antibody therapy. AB - In this study, we compare various methods for the detection of a tumor-associated target antigen and deposition of the bound therapeutic monoclonal antibody in patients enrolled in two separate trials, one involving the administration of two radiolabeled monoclonal antibodies and the other involving an unlabeled antibody. In the first trial, patients with TAG-72 expressing metastatic colon cancer scheduled for surgical intervention received radiolabeled murine and chimeric B72.3 antibody followed by radioimmune imaging and subsequent laparotomy. Normal and tumor tissues obtained at surgery were processed for routine histology, immunohistochemistry, radiometry, and autoradiography. Both anti-TAG-72 antibodies localized to known tumor sites as evidenced by radioimmune imaging. Resected tissue revealed a high tumor-to-normal radiolocalization ratio, and autoradiography demonstrated even deposition of the radiolabeled antibodies throughout the entire tumor deposit with sparing of surrounding normal tissue. In contrast, immunohistochemistry on the same sections revealed comparatively weak antigen expression and patchy antibody localization. In the second trial, patients with GD2 antigen expressing metastatic melanoma received the unlabeled chimeric anti-GD2 antibody C14.18. Immunologic detection of the GD2 antigen and C14.18 deposition was performed on biopsy section as well as on single cell suspension. FACS analysis of the single cell suspension proved more sensitive for the detection of bound antibody than immunohistochemistry, although both methods yielded comparable results for GD2 antigen expression. Our findings demonstrate that the optimal method for the detection of tumor-associated antigen and bound therapeutic antibody can vary depending upon the nature of the antibody (radiolabeled vs. unlabeled and murine vs. chimeric), fixation stability of the target antigen, and the type of pathologic material available for study. PMID- 9735878 TI - Demonstration of platinum microcoils in embolized blood vessels in-situ using a modified methyl methacrylate embedding method, and a special cutting and grinding technique. AB - With current paraffin embedding techniques it is not possible to demonstrate metal particles in vascular lumens. We used a modified methyl methacrylate embedding method for obtaining plastic blocks which were cut with a precision diamond saw, and subsequently ground and polished to preserve the interface of reactive tissue surrounding platinum microcoils in arteriovenous malformations. Artifact-free interfaces could be achieved by slow embedding under vacuum conditions, slow polymerization in a water bath, and very gentle cutting, grinding and polishing techniques. The method produces good preservation of tissue and cells directly adjacent to metal coils. Our method is useful for the histomorphological interpretation of vascular pathologies in which metal particles have been introduced. PMID- 9735879 TI - Effects of sex steroids on silver stained proteins of nucleolar organizer regions (Ag-NOR) in the rabbit uterus. AB - Silver staining of argyrophilic proteins of nucleolar organizer regions (Ag-NOR) has been used to evaluate a hyperactive state of cells. We studied the effects of sex steroids on the Ag-NOR proteins in the rabbit uterus. Estradiol-17beta (E2) caused uterine hypertrophy and increased the number of Ag-NOR dots in epithelial and stromal cells of the endometrium and smooth muscles cells in a dose dependent manner without proliferative change in the endometrium. Progestins including progesterone, medroxyprogesterone acetate (MPA), and norethindrone following E2 caused progestational proliferation of the endometrium with an increase in the Ag NOR number to various extents, whereas methyltestosterone, a synthetic androgen, caused only a minimal proliferative change in the endometrium without an increase in the Ag-NOR number. Western blot analysis revealed that progesterone and MPA increased the amounts of a 100 kDa protein (nucleolin) and a 37 kDa protein (B23 protein) in the endometium and a marked proliferative change, whereas E2 did not. These results suggest that although the Ag-NOR number was enhanced by both progestin and estrogen, only progestins substantially increase the Ag-NOR protein in the rabbit uterus. This implies that an increase in the amount of nucleolin and B23 protein is associated with the proliferative potential of the cells. PMID- 9735880 TI - Neutral red assay modification to prevent cytotoxicity and improve reproducibility using E-63 rat skeletal muscle cells. AB - Cellular uptake of neutral red dye (NR) is currently used as an indirect measure of viable cells in cultures. We used E-63 rat skeletal muscle cells to identify causes of NR assay variability and to develop modifications that substantially reduce it. Three methods of NR preparation and/or addition to cells were used. When NR medium was prepared, incubated overnight, and filtered to remove precipitates, the amount of dye precipitated varied greatly. Coefficients of variation (CVs) in NR uptake were greater than 25% between assays. Higher NR concentrations, longer incubation times, increased pH, and decreased temperature promoted NR precipitation in media. NR media prepared and filtered just prior to use or direct addition of prefiltered NR stock solution to cell cultures resulted in much smaller CVs between assays. NR was cytotoxic to E-63 rat muscle and primary quail myoblasts in a time- and concentration-dependent manner. NR exposure to E-63 cells for greater than 1.25 and 2 hr at 157 or 127 microg/ml, respectively, was associated with swelling and rupture of lysosomes. By contrast, there was no evidence of cytotoxicity when E-63 cells were exposed to NR for 1 hr at either 127 or 157 microg/ml. Primary quail myoblasts developed lysosomal swelling and ruptured more rapidly than E-63 cells when exposed to NR at either 127 or 157 microg/ml. For confluent 10-day cultures of E-63 cells exposed to NR at 127 microg/ml for 1 hr, the CVs within assay and between assays were 3.3-3.9% and 5.1%, respectively. For similarly exposed, actively replicating 3-day cultures of E-63 cells, the CVs within and between assays were 6.2-9.6% and 2.4%, respectively. NR uptake by the E-63 cells was linear with respect to viable cell number. PMID- 9735881 TI - Ruthenium red preserves glycoprotein peplomers of C-type retroviruses for transmission electron microscopy. AB - Peplomers, the glycoprotein projections of the outer viral envelope, are distinctive for many viruses. Peplomers of retroviral C-type particles are fragile and are not preserved in standard preparations for transmission electron microscopy of thin sections, whereas the peplomers of B- and D- type retroviruses are usually preserved. Ruthenium red, extensively used in transmission electron microscopy to enhance the preservation of glycosylated proteins, was used in the preparation of three retrovirus-producing lymphoblastoid cell lines: murine SC-1 cells producing the C-type murine leukemia retrovirus LP-BM5 that causes immunodeficiency, human DG-75 cells producing a murine leukemia retrovirus, and human C5/MJ cells producing human T-cell lymphotropic virus type I (HTLV-I). Fixation of cells was carried out with ruthenium red present in the glutaraldehyde, osmium tetroxide, and the ethanol dehydration through the 70% ethanol step. The detailed structure of peplomers of these three different viruses was well preserved. PMID- 9735882 TI - Improvement of nonradioactive DNA in situ hybridization. AB - With the introduction of microwave pretreatment, the quality of nonradioactive in situ hybridization (NISH) using DNA probes on formalin fixed tissue has significantly improved. Even after microwave treatment, however, there are cases where NISH results remain unsatisfactory. Therefore, we tried to improve NISH by testing other buffer systems as alternatives to the citrate buffer that is routinely applied during microwave pretreatment. By using buffer systems originally designed for immunohistochemistry, we significantly improved our NISH results. Difficult tissue samples were more accessible to NISH using these alternative buffer systems and made the quantitative evaluation easier. These results may also be of interest for combined applications of NISH and immunohistochemistry. PMID- 9735883 TI - Assessment of cerebral pressure autoregulation in humans--a review of measurement methods. AB - Assessment of cerebral autoregulation is an important adjunct to measurement of cerebral blood flow for diagnosis, monitoring or prognosis of cerebrovascular disease. The most common approach tests the effects of changes in mean arterial blood pressure on cerebral blood flow, known as pressure autoregulation. A 'gold standard' for this purpose is not available and the literature shows considerable disparity of methods and criteria. This is understandable because cerebral autoregulation is more a concept rather than a physically measurable entity. Static methods utilize steady-state values to test for changes in cerebral blood flow (or velocity) when mean arterial pressure is changed significantly. This is usually achieved with the use of drugs, shifts in blood volume or by observing spontaneous changes. The long time interval between measurements is a particular concern in many of the studies reviewed. Parallel changes in other critical variables, such as pCO2, haematocrit, brain activation and sympathetic tone, are rarely controlled for. Proposed indices of static autoregulation are based on changes in cerebrovascular resistance, on parameters of the linear regression of flow/velocity versus pressure changes, or only on the absolute changes in flow. The limitations of studies which assess patient groups rather than individual cases are highlighted. Newer methods of dynamic assessment are based on transient changes in cerebral blood flow (or velocity) induced by the deflation of thigh cuffs, Valsalva manoeuvres, tilting and induced or spontaneous oscillations in mean arterial blood pressure. Dynamic testing overcomes several limitations of static methods but it is not clear whether the two approaches are interchangeable. Classification of autoregulation performance using dynamic methods has been based on mathematical modelling, coherent averaging, transfer function analysis, crosscorrelation function or impulse response analysis. More research on reproducibility and inter-method comparisons is urgently needed, particularly involving the assessment of pressure autoregulation in individuals rather than patient groups. PMID- 9735884 TI - Limb capillary filtration coefficient in human subjects: the importance of the site of measurement. AB - Capillary filtration coefficient is a critical determinant of fluid flux across the microvascular wall. Changes in capillary filtration coefficient have been described in a number of disease processes. Measurement is typically made by venous occlusion plethysmography using either the upper or lower limb, but a variety of measurement protocols have been used and the importance of the site of measurement remains unclear. In this study, forearm and calf capillary filtration coefficient were measured in healthy volunteers, either simultaneously (group A; n = 11) or sequentially in random order (group B; n = 11) using venous occlusion plethysmography, with the subject supine and the limb at heart level. In both studies capillary filtration coefficient was significantly higher when measured at the forearm than at the calf (group A: 6.1 +/- 1.0 versus 3.7 +/- 1.1 x 10(-3) ml min(-1) mmHg(-1) 100 ml(-1) (mean +/- SD), p < 0.01; group B: 5.1 +/- 1.2 versus 3.2 +/- 1.1 x 10(-3) ml min(-1) mmHg(-1) 100 ml(-1), p < 0.01). Isovolumetric venous pressure (the maximum pressure at which there is neither net filtration nor absorption at the microvascular wall) was similar in upper and lower limbs in both groups of subjects. We conclude that limb capillary filtration coefficient is dependent on the site of measurement. Caution is required when comparing data recorded at different sites even if corrected for the volume of soft tissue under study. PMID- 9735885 TI - Poststenotic coronary blood flow following percutaneous transluminal coronary angioplasty. AB - Poststenotic intracoronary flow velocity measurements both prior to and following percutaneous transluminal coronary angioplasty (PTCA) by use of a Doppler-tipped guidewire allow estimation of haemodynamic improvement due to interventional procedures. Since poststenotic coronary artery vasoconstriction routinely occurs after PTCA, haemodynamic improvement may be overestimated when measured by flow velocity alone. In 38 patients scheduled for elective PTCA in single vessel disease (left anterior descending = 19; left circumflex = 9; right coronary artery = 10) change of poststenotic coronary blood flow (CBF) was calculated by the combined use of intracoronary flow velocity measurement (average peak velocity: APV) and quantitative coronary angiography (cross sectional area: CSA) both prior to and following PTCA. Poststenotic coronary diameters revealed a small but significant decrease following PTCA (2.9 +/- 0.5 versus 2.7 +/- 0.5 mm, p < 0.001, 33 of 38 analysed vessels, i.e. 86.8%), whereas APV demonstrated a significant increase due to PTCA (17.0 +/- 8 versus 41.5 +/- 16, p < 0.001). Along with the increment in poststenotic flow velocity, poststenotic CBF increased highly significantly following PTCA (33 +/- 25 versus 73 +/- 41 ml min( 1), p < 0.001). In spite of a significant decrease in poststenotic coronary diameter, a highly significant increment of poststenotic flow due to PTCA can be demonstrated paralleling increment of poststenotic coronary Doppler-flow velocity. PMID- 9735886 TI - Estimation of coronary blood flow by ECG gated cardiac thermography in open-chest conditions. AB - Thermography is suggested as a tool to estimate myocardial and coronary epicardial flow in open-chest heart surgery. To test the feasibility and compare various methods for coronary flow estimation in open-chest surgery, thermographic imaging was applied to eight open-chest dogs which were injected with cold saline into the aortic root. Blood flow in the left arterial descending (LAD) coronary vessel was measured by a transit-time flowmeter. ECG gated images were acquired for 20-30 s, while the cold saline (20 ml) was injected into the aortic root. Several flow levels were achieved during repeated hyperaemic response to transient occlusions of the LAD. A temperature response curve for each flow level was obtained by averaging over an edge-detected arterial segment for each image frame. Several indices were calculated from the temperature curve and correlated with the measured coronary flow. These include: an index based on a corrective heat transfer model (r = 0.69, p < 0.001), the slope of the descending part of the response curve (r = 0.76, p < 0.001), the peak temperature difference (r = 0.66, p < 0.001), and the area above the temperature response curve (r = 0.61, p < 0.01). As shown, coronary flow can be estimated quantitatively by intraoperative epicardial thermography, and may therefore provide important on line information regarding blood flow during open-chest surgical procedures. Further studies are required for optimal application of this technique so as to increase its potential as a valid clinical tool. PMID- 9735887 TI - Imaging the structure of the striatum: a fractal approach to SPECT image interpretation. AB - The spatial pattern of striatal dopamine transporter density in the living human brain was tested by duplicate SPECT scans with [123I]PE2I, [123I]beta-CIT or [123I]beta-CIT-FP and striatal phantom measurements. The resolution-dependent spatial variation was calculated by the fractal analysis of SPECT images. This variation, which depends on the size of the region of interest, was described by the spatial dispersion i.e. the standard deviation of the count densities divided by the mean density. In each sub-region, the observed and methodological dispersions were computed, and the resulting spatial dispersion was calculated. The methodological dispersion is caused by the imaging resolution, flood field non-uniformity, count density, scatter, reconstruction errors and partial volume effects, whereas the spatial dispersion is based on the cerebral heterogeneity of the dopamine transporter density. Recognition of the normal variation in heterogeneity is important in evaluation of the striatal dopamine transporter density between controls and patients suffering from various neuropsychiatric disorders. PMID- 9735888 TI - Continuous monitoring of sweating by electrical conductivity measurement. AB - A method of continuous monitoring of sweating was developed in which sweat was detected by changes in the conductivity of perfusing water. An ion-free solution was perfused at a constant flow rate through a chamber attached to the skin surface. The chamber was designed so that the electrodes were installed inside at the inlet and outlet, and a 14 mm3 channel was constructed at the bottom to wash out sweat. The 90% response time was 0.12 s. Attaching the chamber to the palm allowed measurements to be made with the subject seated in a comfortable environment. The sweat rate and heart rate were measured simultaneously with an air-ventilation chamber and a heart rate counter, respectively, with the subjects at rest, and under stresses such as grasping hands and doing mental arithmetic. This method yields sweat responses similar to those obtained with an air ventilation chamber and simultaneous heart rate measurements. The main advantages of this method are faster response time and smaller observation area. PMID- 9735889 TI - Accuracy of movement tracking with a miniaturized second-generation Moire device. AB - The availability of a Nanoform 600, a machine used in the contouring of optical components, with movement capabilities of 12.5 A, made it possible to determine the sensitivity, accuracy and movement tracking ability of a high-resolution measuring device using the principle of Moire magnification. The Nanoform 600 was programmed to make a series of movements measured by the Moire device. Comparison of the Moire measurement with the programmed movements verified that the Moire device is capable of reliably tracking movements as small as 0.05 microm. Increasing the rate of movement had little effect on linearity. Tracking movements over a 350 microm distance at rates of up to 340 microm per second resulted in a 0.23% error. Intentionally defocusing the Moire signal did not appreciably affect sensitivity. PMID- 9735890 TI - Body fat prediction from skinfold anthropometry referenced to a new gold standard: in vivo neutron activation analysis and tritium dilution. AB - Skinfold anthropometry is a widely practiced technique often with little appreciation of its limitations. The large residual error appearing in any regressions of body density versus sums of skinfolds is primarily due to biological causes, in particular the non-constancy of the ratio of subcutaneous to total body fat. Nevertheless this preliminary study shows that the residual error can be reduced by referencing to a gold standard other than body density. Using a difference technique involving in vivo neutron activation analysis and tritiated water dilution, this paper shows that at least in 20-29 year old normal subjects the residual error (expressed as a percentage of mean total body fat in the respective groups) can be reduced from 22% to 16% (p < 0.001) in males and from 17% to 9% in females (p < 0.0001). It is suggested that a large scale study could be initiated with this new gold standard to obtain accurate predictor relationships throughout the whole age range for both sexes. PMID- 9735891 TI - Calibration and assessment of a fluid-filled catheter-transducer system for the measurement of ventricular diastolic pressures. AB - A concise set of experiments is described which detail the calibration of a fluid filled catheter-transducer system and the assessment of a widely used industrial algorithm for determining end-diastolic pressures using that system. First, the static response of the catheter-transducer system was evaluated in vitro by inserting the catheter into a graduated cylinder of saline. Twelve observations revealed a systematic undervaluation of pressure by the system of 1.78 mmHg with 95% limits of agreement ranging from -6.22 to 2.66 mmHg. Next, the dynamic response was evaluated in vivo by performing a transient step-response test. The system had an adequate dynamic response (fn = 11.12 Hz) for intraventricular pressure waveform replication but was considerably underdamped (beta = 0.16). Finally, the ability of the analysis software to detect the point of end-diastole and evaluate end-diastolic pressure was assessed by comparing system output with manual measurements of end-diastolic pressure in 12 patients. The mean difference between manually determined end-diastolic pressure and system output was 0.83 +/- 1.68 mm Hg. This difference is clinically insignificant and shows that the more noteworthy source of error is in the manometer-transducer emphasizing the importance of calibration and quality assurance of fluid-filled catheter transducer systems for use in clinical cardiology or research. PMID- 9735892 TI - Oesophageal transit time evaluated by a biomagnetic method. AB - The aim of this work was to determine the oesophageal transit time (OTT) of a bolus using the biomagnetic technique and compare the results to those obtained by means of scintigraphy. For the biomagnetic evaluation, a test meal (yoghurt) uniformly labelled with 5 g of powder ferrite was swallowed in a single gulp by 19 normal volunteers in the upright position. One sensor (first order gradiometer) was placed at the furcula and a second one at the xiphoid process to detect the passage of the test meal and the magnetic signal output was recorded in a computer. The OTT was determined by plotting the voltage signal against time. The scintigraphic technique was used in the same volunteers: the test meal was labelled with less than 350 MBq of 99mTc-phytate and swallowed in the same way. The bolus transit was recorded at 4 frames s(-1) (100-120 frames acquisition) and the OTT was determined by drawing two regions of interest in the same areas as the sensors. The results were determined by plotting counts against time. The averages for OTTs were 3.8 +/- 0.8 s for the scintigraphic technique and 4.6 +/- 0.9 s for the biomagnetic technique. Although scintigraphic OTT was significantly shorter than magnetic OTT, there was a significant correlation between them. We conclude that the biomagnetic study may be used to evaluate OTT. PMID- 9735893 TI - Perinatal lung function measurements and prediction of respiratory problems in infancy. AB - The aim of this study was to determine which lung function test employed in the perinatal period gave the results most significantly associated with respiratory problems in infancy. The ratio of the proportion of time to reach peak tidal expiratory flow to total expiratory time (tPTEF:tE), thoracic gas volume (TGV) and airway resistance (R(aw)) (from which specific conductance (SG(aw)) was calculated) measurements were examined from 85 infants born at or near term. The infants were followed until at least one year of age and described as symptomatic if they wheezed for at least 24 hours. Twenty-three infants were symptomatic in the first year. The symptomatic group, compared to the asymptomatic, had a higher median FRC (p < 0.01) and R(aw) (p < 0.001); their median SG(aw) was lower (p < 0.001). It was possible to obtain tPTEF:tE results from only 61 infants; the median tPTEF:tE did not differ significantly between symptomatic and asymptomatic infants. Logistic regression analysis demonstrated a high R(aw) and FRC, but not a low tPTEF:tE, independently related to positive symptom status. A high R(aw) (>26 cm H2O (1 s(-1))(-1)) was the most sensitive (83%) predictor of subsequent respiratory problems, but all the tests examined had low positive predictive values. PMID- 9735894 TI - Improved control of gas humidity in neonatal intensive care ventilators. AB - Difficulties experienced by nursing staff in controlling humidity in neonatal intensive care ventilators are shown to originate in the use of a single control, namely a heater, to condition two parameters, namely temperature and humidity, at the inlet to the inspiratory tube. It is shown that better control can be achieved by separating the gas flow through the humidifier into two streams, only one of which is humidified. It is proposed that the temperature along most of the inspiratory tube should be maintained about 3-5 degrees C higher than the target delivery temperature. This eliminates the risk of condensation in the tube, a second commonly occurring problem in existing systems. It is shown why the control of ventilator systems should be based on absolute humidity, not relative humidity, even though the latter may be preferred for monitoring purposes. PMID- 9735895 TI - Measuring human ventilation for apnoea detection using an optical encoder. AB - We have designed, built and tested a proof-of-concept system based on optical encoder technology for measuring adult or infant ventilation. It uses change in chest circumference to provide an indirect measure of ventilation. The Hewlett Packard HEDS-9720 optical encoder senses displacement of its matching codestrip. It yields a resolution of 0.17 mm and is accurate to 0.008 mm over a 10 mm test distance. The encoder is mounted on a nylon web belt wrapped around the torso and responds to changes in circumference. Motion of the code strip during respiration is converted to direction of movement (inhalation or exhalation) as well as magnitude of circumference change. Use of two sensor bands, one on the chest and one on the abdomen, may allow detection of obstructive apnoea in which there is no air flow out of or into the subject despite respiratory movement. Applications of this technology include infant apnoea monitoring as well as long-term adult monitoring. PMID- 9735896 TI - Application of reconstruction-based scatter compensation to thallium-201 SPECT: implementations for reduced reconstructed image noise. AB - Scatter compensation in Tl-201 single photon emission computed tomography (SPECT) presents an interesting challenge because of the multiple emission energies and relatively large proportion of scattered photons. In this paper, we present a simulation study investigating reconstructed image noise levels arising from various implementations of iterative reconstruction-based scatter compensation (RBSC) in Tl-201 SPECT. A two-stage analysis was used to study single and multiple energy window implementations of reconstruction-based scatter compensation, and RBSC was compared to the upper limits on performance for other approaches to handling scatter. In the first stage, singular value decomposition of the system transfer matrix was used to analyze noise levels in a manner independent of the choice of reconstruction algorithm, providing results valid across a wide range of regularizations. In the second stage, the data were reconstructed using maximum-likelihood expectation-maximization, and the noise properties of the resultant images were analyzed. The best RBSC performance was obtained using multiple energy windows, one for each emission photopeak, and RBSC outperformed the upper limit on subtraction-based compensation methods. Implementing RBSC with the correct choice of energy window acquisition scheme is a promising method for performing scatter compensation for Tl-201 SPECT. PMID- 9735897 TI - Optimized acquisition time and image sampling for dynamic SPECT of Tl-201. AB - With the recent development in scatter and attenuation correction algorithms, dynamic single photon emission computerized tomography (SPECT) can potentially yield physiological parameters, with tracers exhibiting suitable kinetics such as thallium-201 (Tl-201). A systematic way is proposed to investigate the minimum data acquisition times and sampling requirements for estimating physiological parameters with quantitative dynamic SPECT. Two different sampling schemes were investigated with Monte Carlo simulations: 1) Continuous data collection for total study duration ranging from 30-240 min. 2) Continuous data collection for first 10-45 min followed by a delayed study at approximately 3 h. Tissue time activity curves with realistic noise were generated from a mean plasma time activity curve and rate constants (K1 - k4) derived from Tl-201 kinetic studies in 16 dogs. Full dynamic sampling schedules (DynSS) were compared to optimum sampling schedules (OSS). We found that OSS can reliably estimate the blood flow related K1 and Vd comparable to DynSS. A 30-min continuous collection was sufficient if only K1 was of interest. A split session schedule of a 30-min dynamic followed by a static study at 3 h allowed reliable estimation of both K1 and Vd avoiding the need for a prolonged (>60-min) continuous dynamic acquisition. The methodology developed should also be applicable to optimizing sampling schedules for other SPECT tracers. PMID- 9735898 TI - Coupled B-snake grids and constrained thin-plate splines for analysis of 2-D tissue deformations from tagged MRI. AB - Magnetic resonance imaging (MRI) is unique in its ability to noninvasively and selectively alter tissue magnetization and create tagged patterns within a deforming body such as the heart muscle. The resulting patterns define a time varying curvilinear coordinate system on the tissue, which we track with coupled B-snake grids. B-spline bases provide local control of shape, compact representation, and parametric continuity. Efficient spline warps are proposed which warp an area in the plane such that two embedded snake grids obtained from two tagged frames are brought into registration, interpolating a dense displacement vector field. The reconstructed vector field adheres to the known displacement information at the intersections, forces corresponding snakes to be warped into one another, and for all other points in the plane, where no information is available, a C1 continuous vector field is interpolated. The implementation proposed in this paper improves on our previous variational-based implementation and generalizes warp methods to include biologically relevant contiguous open curves, in addition to standard landmark points. The methods are validated with a cardiac motion simulator, in addition to in-vivo tagging data sets. PMID- 9735899 TI - Maximum-likelihood estimation of Rician distribution parameters. AB - The problem of parameter estimation from Rician distributed data (e.g., magnitude magnetic resonance images) is addressed. The properties of conventional estimation methods are discussed and compared to maximum-likelihood (ML) estimation which is known to yield optimal results asymptotically. In contrast to previously proposed methods, ML estimation is demonstrated to be unbiased for high signal-to-noise ratio (SNR) and to yield physical relevant results for low SNR. PMID- 9735900 TI - Algebraic reconstruction for magnetic resonance imaging under B0 inhomogeneity. AB - In magnetic resonance imaging, spatial localization is usually achieved using Fourier encoding which is realized by applying a magnetic field gradient along the dimension of interest to create a linear correspondence between the resonance frequency and spatial location following the Larmor equation. In the presence of B0 inhomogeneities along this dimension, the linear mapping does not hold and spatial distortions arise in the acquired images. In this paper, the problem of image reconstruction under an inhomogeneous field is formulated as an inverse problem of a linear Fredholm equation of the first kind. The operators in these problems are estimated using field mapping and the k-space trajectory of the imaging sequence. Since such inverse problems are known to be ill-posed in general, robust solvers, singular value decomposition and conjugate gradient method, are employed to obtain corrected images that are optimal in the Frobenius norm sense. Based on this formulation, the choice of the imaging sequence for well-conditioned matrix operators is discussed, and it is shown that nonlinear k space trajectories provide better results. The reconstruction technique is applied to sequences where the distortion is more severe along one of the image dimensions and the two-dimensional reconstruction problem becomes equivalent to a set of independent one-dimensional problems. Experimental results demonstrate the performance and stability of the algebraic reconstruction methods. PMID- 9735901 TI - Application of the extremum stack to neurological MRI. AB - The extremum stack, as proposed by Koenderink, is a multiresolution image description and segmentation scheme which examines intensity extrema (minima and maxima) as they move and merge through a series of progressively isotropically diffused images known as scale space. Such a data-driven approach is attractive because it is claimed to be a generally applicable and natural method of image segmentation. The performance of the extremum stack is evaluated here using the case of neurological magnetic resonance imaging data as a specific example, and means of improving its performance proposed. It is confirmed experimentally that the extremum stack has the desirable property of being shift-, scale-, and rotation-invariant, and produces natural results for many compact regions of anatomy. It handles elongated objects poorly, however, and subsections of regions may merge prematurely before each region is represented as a single node. It is shown that this premature merging can often be avoided by the application of either a variable conductance-diffusing preprocessing step, or more effectively, the use of an adaptive variable conductance diffusion method within the extremum stack itself in place of the isotropic Gaussian diffusion proposed by Koenderink. PMID- 9735902 TI - Reversible decorrelation method for progressive transmission of 3-D medical image. AB - In this paper, we present a new reversible decorrelation method of three dimensional (3-D) medical images for progressive transmission. Progressive transmission of an image permits gradual improvement of image quality while being displayed. When the amount of image data is very large, as a 3-D medical image, the progressive transmission plays an important role in viewing or browsing the image. The data structure presented in this paper takes account of interframe correlation as well as intraframe correlation of the 3-D image. This type of data structure has been termed the 3-D hierarchy embedded differential image (3-D HEDI) as was derived from the earlier HEDI structure. Experiments were conducted to verify the performance of 3-D HEDI in terms of the decorrelation efficiency as well as the progressive transmission efficiency. It is compared with those of conventional hierarchy interpolation (HINT), two-dimensional (2-D) HEDI and differential pulse code modulation (DPCM). Experimental results indicate that 3-D HEDI outperforms HINT, 2-D HEDI and DPCM in both decorrelation efficiency as well as the progressive transmission efficiency on 3-D medical images. PMID- 9735903 TI - Quasi-bandlimited properties of radon transforms and their implications for increasing angular sampling densities. AB - The n-dimensional (n-D) radon transform, which forms the mathematical basis for a broad variety of tomographic imaging applications, can be viewed as an n-D function in n-D sinogram space. Accurate reconstruction of continuous or discrete tomographic images requires full knowledge of the radon transform in the corresponding n-D sinogram space. In practice, however, one can have only a finite set of discrete samples of the radon transform in the sinogram space. One often derives the desired full knowledge of the radon transform from its discrete samples by invoking various interpolation algorithms. According to the Wittaker Shannon sampling theorem, a necessary condition for a full and unique recovery of the radon transform from its discrete samples is that the radon transform itself be bandlimited. Therefore, it is necessary to analyze the bandlimited properties of the radon transform. In this work, we analyze explicitly the bandlimited properties of the radon transform and show that the radon transform is mathematically quasi-bandlimited [or essentially bandlimited] in two quantitative senses and can essentially be treated as bandlimited in practice. The quasi bandlimited properties can be used for increasing the angular sampling density of the radon transform. PMID- 9735904 TI - Anatomic region-based dynamic range compression for chest radiographs using warping transformation of correlated distribution. AB - The purpose of this paper is to investigate the effectiveness of our novel dynamic range compression (DRC) for chest radiographs. The purpose of DRC is to compress the gray scale range of the image when using narrow dynamic range viewing systems such as monitors. First, an automated segmentation method was used to detect the lung region. The combined region of mediastinum, heart, and subdiaphragm was defined based on the lung region. The correlated distributions, between a pixel value and its neighboring averaged pixel value, for the lung region and the combined region were calculated. According to the appearance of overlapping of two distributions, the warping function was decided. After pixel values were warped, the pixel value range of the lung region was compressed while preserving the detail information, because the warping function compressed the range of the averaged pixel values while preserving the pixel value range for the pixels which had had the same averaged pixel value. The performance was evaluated with our criterion function which was the contrast divided by the moment, where the contrast and the moment represent the sum of the differences between the pixel values and the averaged values of eight pixels surrounding that pixel, and the sum of the differences between the pixel values and the averaged value of all pixels in the region-of-interest, respectively. For 71 screening chest images from Johns Hopkins University Hospital (Baltimore, MD), this method improved our criterion function at 11.7% on average. The warping transformation algorithm based on the correlated distribution was effective in compressing the dynamic range while simultaneously preserving the detail information. PMID- 9735905 TI - A robust numerical solution to reconstruct a globally relative shear modulus distribution from strain measurements. AB - To noninvasively quantify tissue elasticity for differentiating malignancy of soft tissue, we previously proposed a two-dimensional (2-D) mechanical inverse problem in which simultaneous partial differential equations (PDE's) represented the target distribution globally of relative shear moduli with respect to reference shear moduli such that the relative values could be determined from strain distributions obtained by conventional ultrasound (US) or nuclear magnetic resonance (NMR) imaging-based analysis. Here, we further consider the analytic solution in the region of interest, subsequently demonstrating that the problem is inevitably ill-conditioned in real-world applications, i.e., noise in measurement data and improper configurations of mechanical sources/reference regions make it impossible to guarantee the existence of a stable and unique target global distribution. Next, based on clarification of the inherent problematic conditions, we describe a newly developed numerical-based implicit integration approach that novelly incorporates a computationally efficient regularization method designed to solve this differential inverse problem using just low-pass filtered spectra derived from strain measurements. To evaluate method effectiveness, reconstructions of the global distribution are carried out using intentionally created ill-conditioned models. The resultant reconstructions indicate the robust solution is highly suitable, while also showing it has high potential to be applied in the development of an effective yet versatile diagnostic tool for quantifying the distribution of elasticity in various soft tissues. PMID- 9735906 TI - Model-guided labeling of coronary structure. AB - Assigning anatomic labels to coronary arteries in X-ray angiograms is an important task in medical imaging, motivated by the desire to standardize the assessment of coronary artery disease and to facilitate the three-dimensional (3 D) reconstruction and visualization of the coronary vasculature. However, automatic labeling poses a number of significant challenges, including the presence of noise, artifacts, competing structures, misleading visual cues, and other difficulties associated with a dynamic and inherently complex structure. We have developed a model-guided approach that addresses these challenges and automatically labels the vascular structure in coronary angiographic images. The approach consists of two models: 1) a symbolic model, represented through a directed acyclic graph, that captures vascular tree hierarchies and branch interrelationships and 2) a generalized 3-D model that captures spatial and geometric relationships. Importantly, the approach detects ambiguities (such as vessel overlaps) that may be found in a frame of a cine sequence, and resolves these ambiguities by considering the information derived from other (unambiguous) frames in the temporal sequence, employing dynamic programming methods to match the image features found in the different (ambiguous and unambiguous) frames. This paper presents this model-guided labeling algorithm and discusses the experimental results obtained from implementing and applying the resulting labeling system to a variety of clinical images. The results indicate the feasibility of achieving robust and consistently accurate image labeling through this model-guided, temporal disambiguation method. PMID- 9735907 TI - A novel approach to microcalcification detection using fuzzy logic technique. AB - Breast cancer continues to be a significant public health problem in the United States. Approximately, 182,000 new cases of breast cancer are diagnosed and 46,000 women die of breast cancer each year. Even more disturbing is the fact that one out of eight women in the United States will develop breast cancer at some point during her lifetime. Since the cause of breast cancer remains unknown, primary prevention becomes impossible. Computer-aided mammography is an important and challenging task in automated diagnosis. It has great potential over traditional interpretation of film-screen mammography in terms of efficiency and accuracy. Microcalcifications are the earliest sign of breast carcinomas and their detection is one of the key issues for breast cancer control. In this study, a novel approach to microcalcification detection based on fuzzy logic technique is presented. Microcalcifications are first enhanced based on their brightness and nonuniformity. Then, the irrelevant breast structures are excluded by a curve detector. Finally, microcalcifications are located using an iterative threshold selection method. The shapes of microcalcifications are reconstructed and the isolated pixels are removed by employing the mathematical morphology technique. The essential idea of the proposed approach is to apply a fuzzified image of a mammogram to locate the suspicious regions and to interact the fuzzified image with the original image to preserve fidelity. The major advantage of the proposed method is its ability to detect microcalcifications even in very dense breast mammograms. A series of clinical mammograms are employed to test the proposed algorithm and the performance is evaluated by the free-response receiver operating characteristic curve. The experiments aptly show that the microcalcifications can be accurately detected even in very dense mammograms using the proposed approach. PMID- 9735908 TI - Data-driven homologue matching for chromosome identification. AB - Karyotyping involves the visualization and classification of chromosomes into standard classes. In "normal" human metaphase spreads, chromosomes occur in homologous pairs for the autosomal classes 1-22, and X chromosome for females. Many existing approaches for performing automated human chromosome image analysis presuppose cell normalcy, containing 46 chromosomes within a metaphase spread with two chromosomes per class. This is an acceptable assumption for routine automated chromosome image analysis. However, many genetic abnormalities are directly linked to structural or numerical aberrations of chromosomes within the metaphase spread. Thus, two chromosomes per class cannot be assumed for anomaly analysis. This paper presents the development of image analysis techniques which are extendible to detecting numerical aberrations evolving from structural abnormalities. Specifically, an approach to identifying "normal" chromosomes from selected class(es) within a metaphase spread is presented. Chromosome assignment to a specific class is initially based on neural networks, followed by banding pattern and centromeric index criteria checking, and concluding with homologue matching. Experimental results are presented comparing neural networks as the sole classifier to our homologue matcher for identifying class 17 within normal and abnormal metaphase spreads. PMID- 9735909 TI - Design and construction of a realistic digital brain phantom. AB - After conception and implementation of any new medical image processing algorithm, validation is an important step to ensure that the procedure fulfills all requirements set forth at the initial design stage. Although the algorithm must be evaluated on real data, a comprehensive validation requires the additional use of simulated data since it is impossible to establish ground truth with in vivo data. Experiments with simulated data permit controlled evaluation over a wide range of conditions (e.g., different levels of noise, contrast, intensity artefacts, or geometric distortion). Such considerations have become increasingly important with the rapid growth of neuroimaging, i.e., computational analysis of brain structure and function using brain scanning methods such as positron emission tomography and magnetic resonance imaging. Since simple objects such as ellipsoids or parallelepipedes do not reflect the complexity of natural brain anatomy, we present the design and creation of a realistic, high resolution, digital, volumetric phantom of the human brain. This three dimensional digital brain phantom is made up of ten volumetric data sets that define the spatial distribution for different tissues (e.g., grey matter, white matter, muscle, skin, etc.), where voxel intensity is proportional to the fraction of tissue within the voxel. The digital brain phantom can be used to simulate tomographic images of the head. Since the contribution of each tissue type to each voxel in the brain phantom is known, it can be used as the gold standard to test analysis algorithms such as classification procedures which seek to identify the tissue "type" of each image voxel. Furthermore, since the same anatomical phantom may be used to drive simulators for different modalities, it is the ideal tool to test intermodality registration algorithms. The brain phantom and simulated MR images have been made publicly available on the Internet (http://www.bic.mni.mcgill.ca/brainweb). PMID- 9735910 TI - Edge detection in medical images using a genetic algorithm. AB - An algorithm is developed that detects well-localized, unfragmented, thin edges in medical images based on optimization of edge configurations using a genetic algorithm (GA). Several enhancements were added to improve the performance of the algorithm over a traditional GA. The edge map is split into connected subregions to reduce the solution space and simplify the problem. The edge-map is then optimized in parallel using incorporated genetic operators that perform transforms on edge structures. Adaptation is used to control operator probabilities based on their participation. The GA was compared to the simulated annealing (SA) approach using ideal and actual medical images from different modalities including magnetic resonance imaging (MRI), computed tomography (CT), and ultrasound. Quantitative comparisons were provided based on the Pratt figure of merit and on the cost-function minimization. The detected edges were thin, continuous, and well localized. Most of the basic edge features were detected. Results for different medical image modalities are promising and encourage further investigation to improve the accuracy and experiment with different cost functions and genetic operators. PMID- 9735911 TI - MR image texture analysis applied to the diagnosis and tracking of Alzheimer's disease. AB - We assess the value of magnetic resonance (MR) image texture in Alzheimer's disease (AD) both as a diagnostic marker and as a measure of progression. T1 weighted MR scans were acquired from 40 normal controls and 24 AD patients. These were split into a training set (20 controls, 10 AD) and a test set (20 controls, 14 AD). In addition, five control subjects and five AD patients were scanned repeatedly over several years. On each scan a texture feature vector was evaluated over the brain; this consisted of 260 measures derived from the spatial gray-level dependence method. A stepwise discriminant analysis was applied to the training set, to obtain a linear discriminant function. In the test set, this function yielded significantly different values for the control and AD groups (p < 10(-4)) with only small group overlap; a classification rate of 91% was obtained. For the repeatedly scanned control subjects, the median increment in the discriminant function between successive scans of 0.12 was not significantly different from zero (p > 0.05); for the repeatedly scanned AD patients the corresponding median increment of 1.4 was significantly different from zero (p < 0.05). MR image texture may be a useful aid in the diagnosis and tracking of Alzheimer's disease. PMID- 9735912 TI - Magnetocardiographic localization of arrhythmia substrates: a methodology study with accessory pathway ablation as reference. AB - In magnetocardiographic (MCG) localization of arrhythmia substrates, a model of the thorax as volume conductor is a crucial component of the calculations. In this study, we investigated different models of the thorax, to determine the most suitable to use in the computations. Our methods and results are as follows. We studied 11 patients with overt Wolff-Parkinson-White syndrome, scheduled for catheter ablation. The MCG registrations were made with a 37-channel "superconducting quantum interference device" system. The underlying equivalent current dipole was computed for the delta-wave. Three models of the thorax were used: the infinite halfspace, a sphere and a box. For anatomical correlation and to define the suitable sphere and box, magnetic resonance images were obtained. As reference we used the position of the tip of the catheter, at successful radio frequency-ablation, documented by cine-fluoroscopy. Nine patients could be evaluated. The mean errors (range) when using the infinite halfspace, the sphere and the box were 96 (49-125), 21 (5-39), and 36 mm (20-58 mm), respectively (p < 0.0001). In conclusion, the sphere was significantly better suited than the other models tested in this study, but even with this model the accuracy of MCG localization must further improve to be clinically useful. More realistic models of the thorax are probably required to achieve this goal. PMID- 9735913 TI - Linear and neural models for classifying breast masses. AB - Computational methods can be used to provide an initial screening or a second opinion in medical settings and may improve the sensitivity and specificity of diagnoses. In the current study, linear discriminant models and artificial neural networks are trained to detect breast cancer in suspicious masses using radiographic features and patient age. Results on 139 suspicious breast masses (79 malignant, 60 benign, biopsy proven) indicate that a significant probability of detecting malignancies can be achieved at the risk of a small percentage of false positives. Receiver operating characteristic (ROC) analysis favors the use of linear models, however, a new measure related to the area under the ROC curve (AZ) suggests a possible benefit from hybridizing linear and nonlinear classifiers. PMID- 9735914 TI - Neospora caninum infection in a Bernese cattle dog from Italy. AB - A cutaneous nodule associated with Neospora caninum infection was diagnosed in a 5-year-old male Bernese cattle dog from Italy. The ulcerative lesion was 2-3 cm wide located in the skin of the tarsal region. Haematological values were normal and the dog did not show any neurological abnormalities. The dermal lesion consisted of a diffuse necrotic dermatitis with a dense infiltrate of mostly neutrophils and macrophages, surrounded by a fibrous wall. Histological sections revealed numerous tachyzoites of N. caninum scattered throughout the tissue. Diagnosis was confirmed both by immunohistochemical staining and electron microscopic examination. The dog had a 1:640 IFAT titre to N. caninum. Four weeks after surgical excision new subcutaneous nodules reappeared. The cutaneous lesions resolved following 21 days of therapy with clindamycin hydrochloride. These observations demonstrate the presence of N. caninum in Italy and confirm that neosporosis should be considered in the differential diagnosis of pyogranulomatous dermatitis in dogs. Clindamycin may be an effective treatment for cutaneous neosporosis. PMID- 9735915 TI - A geographic information system on the potential distribution and abundance of Fasciola hepatica and F. gigantica in east Africa based on Food and Agriculture Organization databases. AB - An adaptation of a previously developed climate forecast computer model and digital agroecologic database resources available from FAO for developing countries were used to develop a geographic information system risk assessment model for fasciolosis in East Africa, a region where both F. hepatica and F. gigantica occur as a cause of major economic losses in livestock. Regional F. hepatica and F. gigantica forecast index maps were created. Results were compared to environmental data parameters, known life cycle micro-environment requirements and to available Fasciola prevalence survey data and distribution patterns reported in the literature for each species (F. hepatica above 1200 m elevation, F. gigantica below 1800 m, both at 1200-1800 m). The greatest risk, for both species, occurred in areas of extended high annual rainfall associated with high soil moisture and surplus water, with risk diminishing in areas of shorter wet season and/or lower temperatures. Arid areas were generally unsuitable (except where irrigation, water bodies or floods occur) due to soil moisture deficit and/or, in the case of F. hepatica, high average annual mean temperature >23 degrees C. Regions in the highlands of Ethiopia and Kenya were identified as unsuitable for F. gigantica due to inadequate thermal regime, below the 600 growing degree days required for completion of the life cycle in a single year. The combined forecast index (F. hepatica+F. gigantica) was significantly correlated to prevalence data available for 260 of the 1220 agroecologic crop production system zones (CPSZ) and to average monthly normalized difference vegetation index (NDVI) values derived from the advanced very high resolution radiometer (AVHRR) sensor on board the NOAA polar-orbiting satellites. For use in Fasciola control programs, results indicate that monthly forecast parameters, developed in a GIS with digital agroecologic zone databases and monthly climate databases, can be used to define the distribution range of the two Fasciola species, regional variations in intensity and seasonal transmission patterns at different sites. Results further indicate that many of the methods used for crop productivity models can also be used to define the potential distribution and abundance of parasites. PMID- 9735916 TI - A geographic information system forecast model for strategic control of fasciolosis in Ethiopia. AB - A geographic information system (GIS) forecast model based on moisture and thermal regime was developed to assess the risk of Fasciola hepatica, a temperate species, and its tropical counterpart, Fasciola gigantica, in Ethiopia. Agroecological map zones and corresponding environmental features that control the distribution and abundance of the disease and its snail intermediate hosts were imported from the Food and Agriculture Organization (FAO) Crop Production System Zones (CPSZ) database on east Africa and used to construct a GIS using ATLAS GIS 3.0 software. Base temperatures of 10 degrees C and 16 degrees C were used for F. hepatica and F. gigantica, respectively, to calculate growing degree days in a previously developed climate forecast system that was modified to allow use of monthly climate data values. The model was validated by comparison of risk indices and environmental features to available survey data on fasciolosis. Monthly Fasciola risk indices of four climatic regions in Ethiopia were used to project infection transmission patterns under varying climatic conditions and strategic chemotherapeutic fasciolosis control schemes. Varying degrees of F. hepatica risk occurred in most parts of the country and distinct regional F. hepatica transmission patterns could be identified. In the humid west, cercariae shedding was predicted to occur from May to October. In the south it occurred from April to May and September to October, depending on the annual abundance of rain. In the north-central and central regions, risk was highest during heavy summer rains and pasture contamination with metacercariae was predicted to occur during August-September, except in wet years, when it may start as early as July and extend up to October. At cooler sites above altitude of 2800 m, completion of an infection cycle may require more than a year. Fasciola gigantica risk was present in the western, southern and north-central regions of the country at altitudes of 1440-2560 m. However, a transmission cycle could be completed in a single year only at elevations below 1700 m. The greatest risk of F. gigantica infection was in the humid western region. Regional strategic chemotherapy schemes of two or three treatments per year were developed. Results suggest that the model can be extrapolated to all CPSZ in the country and adapted for use in control of other vector-borne diseases of economic and public health importance. PMID- 9735917 TI - Plasma aspartate aminotransferase (AST), glutamate dehydrogenase (GLDH) and gamma glutamyl transpeptidase (GGT) activities in water buffaloes with experimental subclinical fasciolosis. AB - The effect of chronic Fasciola hepatica infection on the activity of plasma aspartate aminotransferase (AST), glutamate dehydrogenase (GLDH) and gamma glutamyl transpeptidase (GGT) was investigated in water buffaloes dosed daily with 60 F. hepatica metacercariae over 20 days. Experimental fluke infection caused no clinical signs but provoked an increase in plasma level of IgG directed against F. hepatica from 4 weeks after infection. There was a significant increase in plasma AST from 6 weeks post-infection. Maximal values were reached at 14 weeks and remained significantly elevated by 23 weeks. Plasma GLDH was significantly elevated from 6 to 21 weeks post-infection. Significant increases in plasma GGT occurred from 8 to 26 weeks post-infection, reaching maximal values at 15 weeks. This study shows that plasma enzyme activities may be useful in studies of fluke-induced liver damage in water buffaloes. PMID- 9735918 TI - Experimental infection of the human granulocytic ehrlichiosis agent in horses. AB - Human blood collected from two patients from Westchester County, New York with human granulocytic ehrlichia (HGE) infection was inoculated into two ponies. Inoculated ponies developed clinical signs similar to a previous report (Madigan et al., 1995). Histopathological changes involved follicular hyperplasia of lymphoid tissues. HGE DNA was detected by PCR in muscle, fascia, peritoneum, and adrenal gland after the ponies produced a high level of antibodies to HGE. We suggest that HGE may reside in poorly vascularized connective tissues, where the antibodies may have some difficulties to penetrate, resulting in persistent infection. Since HGE and E. equi cause very similar diseases in both humans and horses, they may be the same organism with minor genetic differences. PMID- 9735919 TI - Canine heartworm (Dirofilaria immitis) detected in red foxes (Vulpes vulpes) in urban Melbourne. AB - Canine heartworm (Dirofilaria immitis) was detected by antigen ELISA in 8 (6.4%) of 125 red foxes (Vulpes vulpes) captured in the Melbourne (Australia) metropolitan area. Circulating microfilariae were also detected by whole blood filtration in six of these foxes. Cursory sampling of mosquitos at two of the sites where positive foxes were captured revealed the existence of two known mosquito vectors of Dirofilaria. The existence of widespread fox populations in Melbourne, together with known mosquito vectors may offer the potential for a sylvatic cycle of canine heartworm within the urban area. PMID- 9735920 TI - Acute blindness associated with monoclonal gammopathy induced by Ehrlichia canis infection. AB - Ehrlichia canis infection was diagnosed in a Labrador retriever presented with a primary complaint of acute blindness. Ocular signs on admission included bilateral hyphema, retinal haemorrhage and retinal detachment. Serum protein electrophoresis results revealed monoclonal gammopathy. This report discusses and suggests the pathogenesis of ocular bleeding in canine monocytic ehrlichiosis. Blood hyperviscosity, elevation in oncotic pressure, vasculitis, thrombocytopenia and platelet dysfunction are all proposed to be important factors in the pathogenesis of acute blindness in canine monocytic ehrlichiosis. PMID- 9735921 TI - Hirudin versus heparin and low-molecular-weight heparin: and the winner is... PMID- 9735922 TI - The discovery of human granulocytotropic ehrlichiosis. AB - Human granulocytotropic ehrlichiosis (HGE) is a tick-borne, acute, nonspecific febrile illness that was first described in 1994. At this writing more than 300 cases have been recognized in areas where the presumed tick vector Ixodes scapularis occurs endemically. Ehrlichiosis is a nonspecific influenza-like illness, and associated laboratory alterations are variable. Characteristic morulae (clusters of bacteria in leukocyte cytoplasm) can frequently be found in the cytoplasm of circulating neutrophils after careful inspection of the peripheral blood smear. The diagnosis is confirmed by demonstrating seroconversion to the HGE agent, positive polymerase chain reaction, or growth of the HGE agent in tissue culture. Doxycycline provides rapid and effective treatment. PMID- 9735923 TI - Hirudin causes more bleeding than heparin in a rabbit ear bleeding model. AB - This study was undertaken to determine the appropriateness of the current practice of using the activated partial thromboplastin time (APTT) to select hirudin doses. A rabbit bleeding ear model was used to compare the effects of various doses of heparin and hirudin on the relationship between the APTT and bleeding. In addition, the effects of these agents on the thrombin clotting time (TCT) and factor Xa clotting time also were examined. Both heparin and hirudin produced a concentration-dependent increase in bleeding. When bleeding was plotted as a function of APTT ratio, even a small increase in APTT ratio within the therapeutic range of 1.5 to 2.5 resulted in a marked increase in bleeding with hirudin but not with heparin. The TCT was more responsive than the APTT or factor Xa clotting time to increases in hirudin-induced bleeding. In this model, hirudin produces more bleeding than heparin when the agents are used in doses that increase the APTT ratio to the same extent. These studies highlight the pitfalls of extrapolating from experience with heparin when choosing a test to monitor new antithrombotics. Our findings also suggest that the TCT may be more responsive than the APTT for monitoring hirudin therapy. PMID- 9735924 TI - Excessive vasoconstriction after stress by the aging kidney: inadequate prostaglandin modulation of increased endothelin activity. AB - The adaptive capacity of the aging kidney to stimulation of the sympathetic nervous system, as induced by a 30-minute mental stress (MS), was assessed in 8 elderly healthy women (68 to 82 years of age) and compared with that of 8 younger women (24 to 40 years of age). The study encompassed 4 consecutive 30-minute periods (baseline, mental stress, recovery 1, and recovery 2). In the elderly subjects, baseline effective renal plasma flow (ERPF)(iodine 131-labeled hippurate clearance) was lower and glomerular filtration rate (GFR)(iodine 125 labeled iothalamate clearance) was proportionally less reduced than in the younger group; the filtration fraction (FF) was higher. The elderly group excreted more endothelin 1 (ET-1) (P < .05), prostaglandin E2 (PGE2), and 6-keto prostaglandin F1alpha (6-keto PGF1alpha)(P < .001 for both)(radioimmunoassay). Mental stress induced similar increases in blood pressure, heart rate, and plasma catecholamines in the 2 age groups, limited to the stimulation period. In the elderly group, mental stress caused a prolonged decrease in ERPF that reached its maximum 60 minutes after mental stress (-33%, P < .05), while GFR remained constant during the whole experiment, so that FF increased. In the younger subjects, renal hemodynamic changes were limited to the mental stress period. ET 1 increased during mental stress and the first recovery period in the elderly group (+50% and +25%, P < .05) as it did in the younger group, but the elderly group differed from the younger in that vasodilating prostaglandins increased only during mental stress. In conclusion, the aging kidney reacts to adrenergic stimulation with more-pronounced and -prolonged vasoconstriction that is probably caused by a defect in prostaglandin modulation of endothelin activity. Autoregulation of GFR is maintained at the expense of increased intraglomerular pressure. PMID- 9735925 TI - Low-density lipoprotein cholesterol can be chemically measured: a new superior method. AB - The association between elevated levels of low-density lipoprotein (LDL) cholesterol and an increased risk of premature coronary heart disease (CHD) is well documented. Most clinical laboratories estimate LDL cholesterol concentrations according to the Friedewald formula. It provides a relatively reliable estimate of LDL cholesterol concentration, provided the triglyceride concentration is <200 mg/dL. However, the reliability is considerably decreased if the triglyceride concentration is > or =400 mg/dL. The interactions between lipoproteins and surfactants, divalent cations, sugars, and lectins were investigated, and we developed a new assay protocol to chemically measure the LDL cholesterol level in serum that does not require immunoseparation or centrifugation. The assay protocol was evaluated by measuring serum samples obtained from 88 patients and 20 healthy volunteers. The triglyceride levels of the patient samples ranged from 66 to 2199 mg/dL, and the samples were classified as <200 mg/dL (n=36) and > or =400 mg/dL (n=52; 23, 3, and 26 patients had type IIb, type III, and type IV hyperlipoproteinemia, respectively) for comparative studies. The accuracy and precision of our assay protocol fulfilled the criteria of the NCEP Lipid Standardization Panel, and no matrix effect influenced the measurements. The assay protocol is less sensitive to LDL-I than to LDL-II and LDL-III. LDL cholesterol measurements correlated well with those obtained by the ultracentrifugal assay of normotriglyceridemic and hypertriglyceridemic samples. This evidence shows that the results obtained with our assay protocol are superior to those obtained with the Friedewald formula. PMID- 9735926 TI - Macrophage inflammatory protein-1alpha C-C chemokine in parapneumonic pleural effusions. AB - Parapneumonic pleural effusions are associated with the presence of a variety of inflammatory cells whose influx into the pleural space is attributed to the presence of inflammatory cytokines. Macrophage inflammatory protein-1alpha (MIP 1alpha), an important mononuclear chemokine, plays a critical role in pulmonary parenchymal inflammatory disease, but its role in the recruitment and activation of mononuclear phagocytes in the pleural space is unknown. In this study we demonstrate that complicated parapneumonic pleural effusions (empyema) and uncomplicated parapneumonic pleural effusions contain significantly (P < .001) higher levels of MIP-1alpha with higher numbers of mononuclear cells when compared with effusions resulting from malignancy and congestive heart failure. The MIP- 1alpha was biologically active and contributed 43% and 37% of the mononuclear chemotactic activity of complicated and uncomplicated parapneumonic pleural fluids, respectively. In vitro, human mesothelial cells, when stimulated with interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha), or bacterial lipopolysaccharide (LPS), produced MIP-1alpha. Northern blot analysis confirmed that both endogenous (IL-1beta or TNF-alpha) and exogenous (LPS) factors induce MIP-1alpha expression in mesothelial cells. Supernatants from activated mesothelial cells demonstrated chemotactic activity for mononuclear cells. This activity was blocked by MIP-1alpha antibody, indicating that the MIP 1alpha released was biologically active. We conclude that in parapneumonic pleural effusions, MIP-1alpha plays a major but not exclusive role in the recruitment of mononuclear leukocytes from the vascular compartment to the pleural space, and pleural mesothelial cells by production of MIP-1alpha actively participate in this process. PMID- 9735927 TI - Urinary loss of immunoglobulin G anti-F(ab)2 and anti-DNA antibody in systemic lupus erythematosus nephritis. AB - The objective of this study was to determine whether the low levels of serum immunoglobulin G (IgG) anti-F(ab)2 seen in some patients with active systemic lupus erythematosus (SLE) were directly related to the deposition of antibody with this specificity in the kidney or alternatively to the urinary loss of IgG anti-F(ab)2. Serum Levels of IgG anti-F(ab)2, anti-tetanus toxoid, and anti-ds DNA antibody were measured in parallel with urinary excretion of these same 3 antibodies in 28 patients with SLE nephritis and in 28 control patients with other forms of chronic kidney disease. Low levels of both serum IgG anti-F(ab)2 or anti-tetanus antibody appeared to correlate with increased levels of urinary loss of these same antibodies in some patients with SLE and in control subjects with kidney disease. However, urinary loss could not account for low serum levels of either IgG antibody in many subjects. Quantitative 24-hour urinary losses of IgG anti-F(ab)2 and anti-DNA were much higher in patients with SLE than in control subjects with kidney disease (P < .05), whereas amounts of IgG urinary loss of anti-tetanus were similar in patients with SLE and in control subjects. In nearly 1 third of SLE nephritis patients, 13% to 53% of total excreted urinary IgG showed anti-DNA enzyme-linked-immunosorbent assay reactivity. Urinary IgG in many patients with SLE showed both anti-DNA and anti-F(ab)2 reactivity, but dual anti-DNA/F(ab)2 specificity was more pronounced in affinity-isolated serum IgG anti-DNA or anti-F(ab)2 than in excreted urinary IgG molecules. The affinity of urinary IgG for either DNA or F(ab)2 was much lower than the same antibody activities measured either in serum or in kidney biopsy eluates. When the relative affinity of anti-DNA antibody in serum, urine, and kidney biopsy eluate was measured in parallel, the highest affinity antibody was found in kidney biopsy eluates, followed by serum antibody with urine antibody affinity showing the lowest values. These findings suggest a relative concentration of the highest affinity, doubly reactive IgG anti-DNA/F(ab)2 in SLE kidney tissues during SLE nephritis and implicate this process as an important factor in ongoing tissue damage. PMID- 9735928 TI - Sparsely adherent platelets support capture and immobilization of flowing neutrophils. AB - Continuous monolayers of activated platelets support selectin-mediated rolling and integrin-mediated immobilization of flowing neutrophils. Because platelets attached to the vessel wall in vivo may not be confluent, we investigated the ability of sparsely adherent platelets to capture flowing neutrophils that were unstimulated or activated with N-formyl-methionyl-leucyl-phenylalanine (fMLP) peptide (10(-7) mol/L). Different degrees of deposition of platelets were obtained by perfusing heparinized blood for varying periods through glass capillaries coated with collagen type III. Perfusion for 10 to 180 seconds gave approximately 1% to approximately 10% coverage of the surface with platelets. When purified neutrophils were subsequently flowed over the platelets at a wall shear stress of 0.1 Pa, they formed repeated short-lived attachments to individual or small groups of platelets. Increasing surface coverage with platelets was associated with increasing numbers of neutrophils adherent at any time, decreasing the overall rate of motion of adherent neutrophils and increasing the stability of adhesion (eg, proportion remaining with time). The net effect was to actually decrease the flux of adherent cells (number of adhesive interactions seen per unit area per unit time), indicating that parameters based on through-flow of adhesive cells must be interpreted with caution. When neutrophils were preactivated with fMLP they formed stable, stationary attachments. The flux of adherent activated cells was low, although the number of cells adherent remained high, reinforcing the conclusion that assays of the adhesion of flowing cells must carefully characterize the different types of adhesive behavior. Overall, small numbers of platelets can support short lived attachments of flowing neutrophils and allow immobilization of activated cells and may amplify inflammatory processes thereby. PMID- 9735929 TI - Renin-aldosterone system can respond to furosemide in patients with hyperkalemic hyporeninism. AB - Thirty-four patients (65.3+/-3.3 years of age, mean+/-SEM) with hyperkalemia (serum potassium >5.0 mEq/L) had measurement of their renin-aldosterone system. Nineteen patients (56%) had plasma renin activity (PRA) >1.5 ng/mL/h, which was not low, while 15 (44%) had PRA <1.5. Twelve of the 15 hyporeninemic hyperkalemic patients were studied to determine whether their renin-aldosterone system responded to 2 weeks of furosemide, 20 mg daily. Four were nonresponders: PRA averaged 0.3+/-0.1 ng/mL/h, and it did not increase with furosemide or respond to captopril before or after furosemide. Eight patients were responders: PRA averaged 0.6+/-0.2 ng/mL/h and increased with furosemide to 5.5+/-3.4 ng/mL/h. Captopril failed to increase PRA before furosemide, but PRA increased to 15.3+/ 8.4 ng/mL/h after furosemide. Plasma aldosterone was low in both nonresponders and responders (3.5+/-1.2 ng/dL vs 5.8+/-2.5 ng/dL) and did not increase significantly with furosemide (4.3+/-1.7 ng/dL vs 8.7+/-2.5 ng/dL). Serum potassium did not fall and therefore did not limit the rise in aldosterone. Renin responders had greater body weight, were predominantly female (6/8 vs 2/4) and were more likely to have diabetes mellitus (7/8 vs 0/4). Plasma atrial natriuretic peptide (ANP) fell with furosemide in 8 of 8 responders and in 1 of the 2 nonresponders in whom it was measured. Neither group had suppressed plasma prorenin levels, indicating no suppression of renin gene expression. These results indicate that many hyperkalemic patients do not have suppressed PRA. Further, a majority of patients with suppressed PRA have high levels of ANP and can respond to diuretic therapy with a rise in PRA and a fall in ANP, suggesting physiologic suppression of the renin system by volume expansion. A minority of hyperkalemic patients with suppressed PRA had PRA that did not increase under these study conditions. PMID- 9735930 TI - Polymorphisms in the dihydrofolate reductase (DHFR) and dihydropteroate synthetase (DHPS) genes of Plasmodium falciparum and in vivo resistance to sulphadoxine/pyrimethamine in isolates from Tanzania. AB - The efficacy of sulphadoxine/pyrimethamine (S/P) in treatment of uncomplicated falciparum malaria in Africa is increasingly compromised by development of resistance. The occurrence of mutations associated with the active site sequence in the Plasmodium falciparum genes coding for dihydrofolate reductase (DHFR) and dihydropteroate synthetase (DHPS) is associated with in vitro resistance to pyrimethamine and sulphadoxine. This study investigates the occurrence of these mutations in infected blood samples taken from Tanzanian children before treatment with S/P and their relationship to parasite breakthrough by day 7. The results show that alleles of DHPS (436-alanine, 437-alanine and 540-lysine) were significantly reduced in prevalence on day 7 after S/P treatment. In this area, a DHPS with 436-serine, 437-glycine and 540-glutamate appears to play a major role in resistance to S/P in vivo. Evidence for the influence of mutations in the DHFR gene in this investigation is not clear, probably because of the high prevalence of 'resistance-related' mutations at day 0 in the local parasite population. For apparently the same reason, it was not possible to show a statistical association between S/P resistance and the presence of particular polymorphisms in the DHFR and DHPS genes before treatment. PMID- 9735931 TI - Acute bacterial meningitis in children admitted to the Queen Elizabeth Central Hospital, Blantyre, Malawi in 1996-97. AB - To design appropriate interventions, we collected clinical and demographic data prospectively on all children aged one day to 14 years admitted with a diagnosis of bacterial meningitis (BM) from April 1st 1996 to March 31st 1997 to the Queen Elizabeth Central Hospital (QECH), Blantyre, Malawi. During the study period 267 children (2.7% of all paediatric admissions) were found to have BM; 83% were under 5 years of age, 61% under one year and 23% under one month. The most common causative organisms in the post neonatal period (n=206) were Streptococcus pneumoniae (27%), Haemophilus influenzae type b (Hib) 21%, and Salmonella typhimurium (6%). In the neonatal group (< 1 month, n=61) the most common causes were Streptococcus agalactiae (23%), S. typhimurium (15%), S. pneumoniae (11.5%) and other Gram negative rods (11.5%). Nineteen of 21 salmonella infections were in children under one year of age and all S. agalactiae were in infants under three months. There was delay on presentation: the average length of fever was 4.6 days, 39.5% had convulsed prior to arrival and 57% had an altered level of consciousness. An initial diagnosis of malaria had probably contributed to the delay in 22.5% (42 of 186 tested). 48% were < 80% weight for age, with 18% < 60%) weight for age. The overall mortality was 40%. The outcome was worst in salmonella infections, particularly neonatal salmonella BM with a case fatality rate (CFR) of 89% (8 of 9 cases). Coma on presentation worsened prognosis (mortality 64% if Blantyre Coma Score < 3, 26% if > 3). 15% of survivors had sequelae on discharge. 20% of Hib isolates were resistant to chloramphenicol, but all salmonellae were sensitive. 5% of S. pneumoniae were resistant to penicillin and 8% to chloramphenicol. Earlier access to adequate health care and awareness of BM in a malaria-endemic area would reduce mortality and morbidity. Vaccination against Hib infection would have reduced death by 18 (17%) and prevented sequelae in 7 cases. PMID- 9735932 TI - A comparison of use of a pyrethroid either for house spraying or for bednet treatment against malaria vectors. AB - In an intensely malarious area in north-east Tanzania, microencapsulated lambdacyhalothrin was used in four villages for treatment of bednets (provided free of charge) and in another four villages the same insecticide was used for house spraying. Another four villages received neither intervention until the end of the trial but were monitored as controls. Bioassays showed prolonged persistence of the insecticidal residues. Light traps and ELISA testing showed reduction of the malaria vector populations and the sporozoite rates, leading to a reduction of about 90% in the entomological inoculation rate as a result of each treatment. Collections of blood fed mosquitoes showed no diversion from biting humans to biting animals. Incidence of re-infection was measured by weekly monitoring of cohorts of 60 children per village, after clearing preexisting infection with chlorproguanil-dapsone. The vector control was associated with a reduction in probability of re-infection per child per week by 54-62%, with no significant difference between the two vector control methods. Cross-sectional surveys for fever, parasitaemia, haemoglobin and weight showed association of high parasitaemia with fever and anaemia and beneficial effects of each intervention in reducing anaemia. However, passive surveillance by resident health assistants showed no evidence for reduced prevalence of fever or parasitaemia. Net treatment consumed only about one sixth as much insecticide as house spraying and it was concluded that the former intervention would work out cheaper and nets were actively demanded by the villagers, whereas spraying was only passively assented to. PMID- 9735933 TI - Recurrence of blackwater fever: triggering of relapses by different antimalarials. AB - Five cases of blackwater fever (BWF) are described, all of whom had a history of recent quinine therapy. In two cases a second haemolytic crisis was induced by halofantrine, in one case also a third. Increasing frequency of this syndrome with its dramatic clinical presentation is to be expected as imported P. falciparum infection, parasite resistance to chloroquine and the use of quinine and other related antimalarials become more frequent. PMID- 9735934 TI - Voluntary health insurance in Bwamanda, Democratic Republic of Congo. An exploration of its meanings to the community. AB - An insurance scheme covering hospital care in the rural district of Bwamanda in the North-west of the Democratic Republic of Congo, which locally is called the mutuelle, was conceived and developed in 1986 on the initiative of Belgian doctors working in the district under the arrangements for bilateral Belgian aid. After more than 10 years of operation the Bwamanda scheme has achieved a high rate of coverage, contributed to a significant improvement in access to hospital based in-patient care, and constitutes a stable source of revenue for the operation of the hospital. We present an investigation conducted through focus groups in 1996 of the population's social perceptions of this risk-sharing scheme to identify ways to improve it. The findings pertain to the reasons for people to subscribe to the scheme; to the perception of its redistribution effects; to people's frustrations and questions; and finally to the relationships between the insurance scheme and traditional mutual aid arrangements. The difference between a hospital insurance scheme (a logic of contract) and the traditional systems of mutual aid (a logic of alliance) is highlighted, and the impact of the hospital insurance scheme on social inequalities is discussed. The implications of this study on the management of the Bwamanda health insurance scheme are reviewed, and this study may be useful to health managers working in similar contexts. PMID- 9735935 TI - Schistosomiasis screening and health education for children: action research in Nile delta villages. AB - In Egypt an important component of the national schistosomiasis control policy is the regular screening of school children through the rural health units. In the Nile delta, a major challenge for the programme is the increasing predominance of Schistosoma mansoni, as compared to Schistosoma haematobium over the past 20 years, and the need to adjust strategies to this new reality. The action research project, growing out of an in-depth study of schistosomiasis in two Nile delta villages, is intended to provide recommendations for schistosomiasis control to the Egyptian Ministry of Health and Population. This paper explores the practice of school-based screening. We then describe the designing and testing of a revised screening procedure, which focuses on the collection of stool specimens to diagnose S. mansoni and which is comprehensive and gender-sensitive. Opportunities for health education are discussed briefly, as this is necessary for an effective screening procedure and to prevent reinfection. In conclusion, we mention the need to incorporate screening and health education into an integrated control strategy PMID- 9735936 TI - The impact of vitamin A supplementation given during a pneumonia episode on the subsequent morbidity of children. AB - OBJECTIVE: To evaluate the impact of large-dose vitamin A supplementation given to infants > 6 months old (200000 IU) and to preschool children aged 1-4 years (400000 IU) during a pneumonia episode, on their subsequent morbidity and severe morbidity. METHOD: In a randomized, double-blind, placebo controlled trial, the children were followed-up with 2-weekly visits at home for 16 weeks, with the first visit 2 weeks after treatment for pneumonia was initiated. The field workers asked about the presence of morbidity on the day of the visit and in the previous two weeks and about the occurrence and number of clinic attendances and hospital admissions since the last visit. They also measured the patients respiratory rate and temperature and assessed the children for the presence of cyanosis, chest indrawing and wheezing. RESULTS: Except for the prevalence of diet refusal which was higher in the vitamin A group, no differences between the study groups were observed, either in the prevalence of morbidity or in the incidence of clinic attendances and hospital admissions. CONCLUSION: No evidence was found for a beneficial effect of vitamin A given during acute pneumonia on the subsequent morbidity and severe morbidity of children in a population with marginal vitamin A deficiency. PMID- 9735937 TI - [Malaria transmission in the rural zone of Niakhar, Senegal]. AB - The anopheline bioecology and the malaria transmission were studied from January to December 1995 in three villages of the sahelian rural area of Niakhar, Senegal. This area of 29000 inhabitants, has been for several decades, a regional observatory for population and health. The three methods used for collecting mosquitoes were the collection at larval stages, the all night human biting collection, and the pyrethrum spray catch in houses during afternoons. The anophelines collected were, by numerical importance: Anopheles arabiensis, An. rufipes, An. gambiae, An. pharoensis, An. funestus and An. coustani. In the An. gambiae complex, An. arabiensis represented 97% of man biting females and 98% of half gravid resting females (difference not significant); the other reminding species of this complex was always An. gambiae. These two species belonging to the An. gambiae complex were responsible for the totality of the transmission. The anthropophilic index, obtained from half gravid indoor resting An. gambiae s.l., was 83%. The annual biting rate of An. gambiae s.l. varied from 512 to 1558 bites per man per night, depending on the villages. Vectors were observed all year long but their densities were low during the dry season. Vector population presented a notable increase due to the rains, with a maximum of about 10 bites per man per night in September or at the beginning of October; during September the biting rate represented 48% of the annual biting rate. The sporozoitic index of An. gambiae s.l., obtained by ELISA revealing the circumsporozoite protein, was 1.6% for human biting females and 1.8% for half-gravid resting females (difference not significant). Plasmodium falciparum was the only plasmodial species observed among infected anophelines. The annual transmission in the vo villages representative of the Niakhar area were 9 and 12 bites of infected anophelines per man, occurring mainly from August to October. In the third village, not representative of the area regarding permanent breeding places, the transmission was 26 bites of infected anopheline per man per year. These results were discussed in the Senegambian and sahelian contexts. PMID- 9735939 TI - Yellow fever vaccination and risk of spontaneous abortion. PMID- 9735938 TI - Nutritional status of children admitted to hospital with different diseases and its relationship to outcome in The Gambia, West Africa. AB - Admission records from two paediatric units in The Gambia were used to explore the relationship between admission weight and different diseases. In total 13579 hospitalized children were analysed. For comparison, 7399 children were recruited from several surveys of well subjects to provide anthropometric values for healthy Gambian children. Compared to the control children, mean admission weights were lower for malaria (weight for age z-score: -1.602), cerebral malaria (-1.547), transfused malarial anaemia (-1.764), pneumonia (-1.725), meningitis ( 1.362), gastro-enteritis (-2.497) and malnutrition (-3.786). Children with bronchiolitis did not have a significantly different weight for age than the controls. Outcome of the hospital admission was recorded and related to the weight on admission. In all disease categories the death rate rose with decreasing admission weight with the exception of bronchiolitis. For all diseases taken together, case fatality was 7.2% for children with a weight for age z-score above -2 Standard Deviations (SD), 9.3% between -2 and -3 SD, 15.6% between -3 and -4 SD and 22.7% for children with weight for age SD z-scores less than -4. Malnourished children are more susceptible to several infectious diseases frequently seen in developing countries and nutritional interventions, as well as standard treatment, may improve outcome. PMID- 9735940 TI - Part I. A look at population-based medical care. AB - Recent trends toward managed health care have generated interest in developing strategies to manage the health care of a population as a whole. Population-based medicine places the individual patient within the context of the larger community, which is composed of both sick and well individuals; when viewed in these terms, only a small proportion of the people who consult a primary care physician are at risk for substantial morbidity. However, the physician serves as the central figure for delivering population-based health care to the entire community. Many strategies for population-based care contain the following 4 basic elements: 1. Identifying the health and disease states that are likely to be responsive to population-based care, 2. Applying principles of epidemiology to define the population-of-interest, 3. Assembling a multidisciplinary team, and 4. Building information systems to support ongoing surveillance of population-based care. To date, most of the published examples of population-based management have been conducted in managed care environments, but population-based management may also be used by a single physician practice or a small group practice. Programs aimed at health promotion or disease prevention are among the easiest to implement. By examining the results of an entire population with a given condition, physicians and their teams may begin to identify ways to improve the overall delivery of care, either by establishing new procedures or improving old ones. PMID- 9735941 TI - Part II. Evidence-based medicine: a passing fancy or the future of primary care? AB - Reliable experimental evidence provides a firm scientific foundation for only a minority of the clinical decisions primary care practitioners must make each day. Thus clinicians' experience and judgment must complement and supplement their knowledge of published research studies. This presents a dynamic and difficult challenge to the practitioner--one that is magnified when combined with the never ending influx of medical information, with patients' and physicians' uneasiness with clinical uncertainty, and with new external pressures to standardize care. With these factors in mind, this article will review evidence-based medicine, a process and philosophy for the practice and teaching of clinical medicine that has sparked much controversy in recent years. Clinical scenarios commonly encountered in adult primary care--acute low back pain, hypertension, and screening for vascular disease--illustrate some strengths and limitations of evidence-based medicine. PMID- 9735942 TI - Part III. Performance measurements of primary care physicians in managed care. AB - A fundamental change occurring for physicians is that there are increasingly organized efforts to comprehensively assess physician performance. Managed care is the factor most instrumental in leading to an enhanced focus on physician measurements. Another major factor that has prompted increased attention to the measurement of physicians' performance is that patients are beginning to act more as consumers of health care. Efforts to measure physician performance in geographically dispersed primary care practices is inherently more difficult than measuring hospital care. However, according to some studies that have attempted to do this, the delivery in primary care offices of basic preventive services and the care given to patients with chronic illnesses is surprisingly poor. If primary care physicians don't address these issues, managed care companies will make it policy to refer some patients with chronic disease to specialists, who are comprehensively achieving higher measurement scores. What is being measured is at present quite variable in different primary care offices. Most of the initial measurements have been from claims data or from other data that might be obtained and aggregated outside of the primary care physician's office. As this data is not very rich in clinical information, significant misinterpretation is possible. In order to augment these shortcomings, office records are increasingly being reviewed. A standardization of primary care physicians' office medical records is rapidly occurring and is being driven by the measurable items reviewed by managed care organizations. Measurement of patient complaints and patient surveys is another means that managed care organizations presently use to assess primary care physicians' performance. Extreme caution should be used when interpreting this data, as often the small numbers of patients, multifactorial issues, and ambiguity about responsible parties may skew the results. Measurement processes are evolving to focus on how the health care system functions in an integrated fashion, instead of strictly on how the individual physician is performing. Present and planned measurement processes should be used to improve care of patients, and this is most likely to occur when physicians take an active role in understanding and responding to the measurement processes arising from managed care. PMID- 9735943 TI - The cerebellum: an overview. AB - Life has been compared to a beautiful tapestry, woven in intricate design of many threads and colors. By means of physics, chemistry, physiology, anatomy, embryology and genetics we unravel this texture, separate its constituent threads and colors, but lose the pattern as a whole. These analytical sciences have enormously increased our knowledge of life's constituent elements and processes, but the pattern of the tapestry is usually neglected or ignored. PMID- 9735944 TI - The anatomy of the cerebellum. AB - Vertebrate cerebella occupy a position in the rostral roof of the 4th ventricle and share a common pattern in the structure of their cortex. They differ greatly in their external form, the disposition of the neurones of the cerebellar cortex and in the prominence of their afferent, intrinsic and efferent connections. PMID- 9735945 TI - The cells and molecules that make a cerebellum. AB - The molecular underpinnings of cerebellar development are being established through the identification of naturally occurring mutated genes and the knockout of other genes. Sets of genes expressed in the regions of the mes- and metencephalon have been shown to play a crucial role in specifying the cerebellar anlage. Other genes have been shown to be crucial to early granule-cell development, migration of Purkinje and granule cells, and neuron-glia interactions. However, the process of development will ultimately be understood in terms of cellular interactions and the roles that each cell type plays in the assembly of cerebellar structure. One of the most important interactions is between granule and Purkinje cells. This relationship has been shown to be crucial for the control of cell number, migration of neuroblasts and cell differentiation. PMID- 9735946 TI - From zebra stripes to postal zones: deciphering patterns of gene expression in the cerebellum. AB - The analysis of patterned gene expression has been an important tool for dissecting the molecular and developmental bases of functional compartmentalization in the mammalian cerebellum. In particular, sagittally oriented cellular aggregates arranged along the mediolateral axis are the patterning element most commonly invoked to illustrate cerebellar compartmentalization, and these are revealed both by patterns of afferent projection and by a number of classical biochemical markers that are distributed in a pattern of'zebra stripes'. Compartmentation along both the mediolateral and rostrocaudal axes might be linked mechanistically to segmentation in the fruit fly, since early cerebellar development is especially dependent upon the expression of mammalian homologs of Drosophila segmentation genes. In addition, as has been demonstrated in the retinotectal system, some of these genes are likely to control positional information required for the sagittal organization of cerebellar afferent projections. However, in contrast to these global or macro zones, the cerebellum is also compartmentalized at the subcellular or micro level. This can be visualized by differential patterns of mRNA distribution within the sole cerebellar efferent system, the Purkinje cell, defining within such cells a number of distinct subcellular domains or 'postal zones'. The global versus subcellular levels of cerebellar compartmentalization are related since they both appear to be linked to patterns of afferent innervation.A major goal of cerebellar research will be to unravel the true nature of such a relationship, and its relevance to function and behavior. PMID- 9735947 TI - Microcircuitry and function of the inferior olive. AB - The inferior olive, which provides the climbing fibers to Purkinje cells in the cerebellar cortex, has been implicated in various functions, such as learning and timing of movements, and comparing intended with achieved movements. For example, climbing-fiber activity could transmit error signals during eye-blink conditioning or adaptation of the vestibulo-ocular reflex, or it could carry motor command signals beating on the rhythm of the oscillating and synchronous firing of ensembles of olivary neurons, or both. In this review, we approach the controversial issue of olivocerebellar function from the perspective of the unique organization of the microcircuitry of the olivary neuropil. The characteristic glomeruli are formed by a core of long dendritic or axonal spines, each of which is innervated by both an inhibitory terminal derived from the hindbrain and an excitatory terminal derived from either an ascending or descending input. The dendritic spines, which originate from dendrites with varicosities carrying dendritic lamellar bodies, are coupled by gap junctions. By drawing a comparison with a computational model by Segev and Rall,which might be applicable to the typical olivary spine with its unique morphological features and combined excitatory and inhibitory input, we propose that the microcircuitry of the inferior olive is capable of functioning both in motor learning and motor timing, but does not directly compare intended with achieved movements. PMID- 9735948 TI - Cellular mechanisms of cerebellar LTD. AB - In the past decade there have been advances in understanding the cellular mechanisms of the long-term depression (LTD) of synaptic transmission at parallel fiber-Purkinje cell synapses in the cerebellum. This review first summarizes current views on mechanisms involved in LTD induction, from activation of voltage gated Ca2+ channels, of ionotropic (AMPA) and metabotropic (mGluRI) glutamate receptors, to stimulation of protein kinase C and nitric oxide formation. Second, we will focus on recent findings that point towards the involvement of Ca2+ release from internal stores in LTD induction, localize the sources and targets of nitric oxide and indicate a postsynaptic site for LTD expression. Finally, a role for LTD in motor learning is now well supported by recent experiments on transgenic mice. PMID- 9735949 TI - Plasticity of the olivocerebellar pathway. AB - The adult olivocerebellar axons and their terminal arbours, the climbing fibres, are capable of remarkable structural plasticity, regulated through their interaction with Purkinje cells. When these cells are deleted,terminal climbing fibre branches retract. In contrast,there is a vigorous outgrowth of entire terminal arbours when extra postsynaptic neurones are available. The new connections lead to a functional, highly specific pattern of innervation at the single Purkinje cell level and are topographically organized according to the principles of the original projection map.A reversible climbing fibre retraction occurs following depression of electrical activity of the cerebellar cortex. These remarkable plastic properties, together with the fact that these neurones express several growth-associated genes constitutively, suggest that the climbing fibre synapses might be adjusted dynamically to participate in physiological plasticity. PMID- 9735951 TI - Profit and quality in managed care. PMID- 9735950 TI - Genes involved in hereditary ataxias. AB - The hereditary ataxias are a group of inherited neurodegenerative disorders characterized by progressive ataxia that results from degeneration of the cerebellum and its afferent and efferent connections. Recent molecular research has led not only to the discovery of a number of causative mutations, but also shed light on the likely mechanisms by which these mutations cause the respective phenotypes. In Friedreich's ataxia (FRDA), the most common type of autosomal recessive ataxia, the loss of a mitochondrial protein, frataxin, results in overload of mitochondrial iron and oxidative stress. The autosomal dominant ataxias, spinocerebellar ataxia type I (SCAI), SCA2, SCA3 and SCA7, are caused by inheritance of an unstable, expanded CAG trinucleotide repeat. These disorders are assumed to be due to a novel deleterious function of the extended polyglutamine sequences within the proteins encoded by the respective genes. Recent observations in transgenic mice and in human post-mortem tissue suggest that the extended proteins are transported into the nucleus of neurons where they form intranuclear inclusions that disrupt normal nuclear function. In another group of dominant disorders, episodic ataxia type I and type 2 (EA-I, EA-2) and SCA6, the mutations affect genes that code for ion channels. PMID- 9735952 TI - Assessment of psychiatric patients' risk of violence toward others. PMID- 9735953 TI - Psychosocial rehabilitation early after the onset of psychosis. PMID- 9735954 TI - Use of mood stabilizers by hospitalized geriatric patients with bipolar disorder. PMID- 9735955 TI - Web sites worth watching. PMID- 9735956 TI - Salary changes among psychologists by gender and years of work. PMID- 9735957 TI - Neuroleptic malignant syndrome: a review. AB - OBJECTIVE: Neuroleptic malignant syndrome is an uncommon side effect of antipsychotic medications characterized by severe rigidity, tremor, fever, altered mental status, autonomic dysfunction, and elevated serum creatinine phosphokinase and white blood cell count. This paper presents a concise and comprehensive review of neuroleptic malignant syndrome, written with the practitioner in mind, to provide information that will be useful in actual clinical settings. METHODS: MEDLINE was searched from 1966 to 1997 for key reviews, reports on series of cases of neuroleptic malignant syndrome, individual case reports, and other clinically and theoretically important information. RESULTS AND CONCLUSIONS: Virtually all neuroleptics are capable of inducing the syndrome, including the newer atypical antipsychotics. The standard of care for the recognition of neuroleptic malignant syndrome has shifted considerably over the past 15 years. Neuroleptic malignant syndrome belongs in the differential diagnosis of any patient receiving a neuroleptic who develops a high fever or severe rigidity. In addition to measurement of creatinine phosphokinase and white blood cell count, important tests to rule out other etiologies include urinalysis to measure electrolytes, including calcium and magnesium; kidney, liver, and thyroid function tests; lumbar puncture; an electroencephalogram; and a computed tomography or magnetic resonance imaging scan of the head. Although specific treatment remains controversial, supportive treatment such as antipyretics, a cooling blanket, and intravenous fluids to correct dehydration and electrolyte abnormalities is critical and widely supported by consensus. Most patients recover from neuroleptic malignant syndrome in two to 14 days without any cognitive impairment, and new dysfunction usually is attributable to very high fever, hypoxia, or other complications, rather than neuroleptic malignant syndrome per se. PMID- 9735958 TI - Inpatient psychiatric treatment of elderly Medicare beneficiaries. AB - OBJECTIVE: The clinical characteristics and treatment patterns of elderly Medicare beneficiaries hospitalized for psychiatric disorders were examined. METHODS: Administrative data on all elderly Medicare beneficiaries in the United States hospitalized in a nonfederal hospital for a primary psychiatric disorder in 1990-1991 were used to calculate descriptive statistics on case-mix by age group, hospital type (psychiatric hospital, general hospital psychiatric unit, or general hospital nonpsychiatric unit), and primary diagnosis. Length of stay, costs, and discharge destination by hospital type and primary diagnosis were also determined. RESULTS: A total of .6 percent of elderly Medicare beneficiaries were hospitalized for a psychiatric disorder in 1990, accounting for more than 240,000 admissions and $1 billion in Medicare payments. The most common reasons for hospitalization were major depressive disorder (28.1 percent), dementia and other organic disorders (26.8 percent), and substance-related disorders (12.6 percent). Organic disorders were particularly prevalent among the oldest old, accounting for more than half of psychiatric admissions among those 85 and older. A total of 43 percent of the psychiatric admissions were to general hospital nonpsychiatric units, 38 percent to general hospital psychiatric units, and only 19 percent to psychiatric hospitals. Within each diagnostic category, patients admitted to general hospital nonpsychiatric units had the shortest average lengths of stay and the lowest average costs. Among beneficiaries with organic, affective, and psychotic disorders other than schizophrenia, those admitted to general hospitals had shorter lengths of stay, higher rates of discharge to nursing homes, and lower rates of discharge to self-care than those treated in psychiatric hospitals. CONCLUSIONS: Case-mix-adjusted treatment patterns varied substantially across hospital types, due to differences in either illness severity or treatment styles. PMID- 9735959 TI - Course of antidepressant treatment drug type, and prescriber's specialty. AB - OBJECTIVE: The study examined whether the relationship between the course of antidepressant treatment and the type of prescriber-psychiatrist or nonpsychiatrist-varied by whether a tricyclic antidepressant or a selective serotonin reuptake inhibitor (SSRI) was prescribed. METHODS: Pharmacy claims from a nationwide database were analyzed retrospectively. A total of 3,101 adults who did not have a prescription for antidepressants for nine months and who were then given a prescription for a tricyclic or an SSRI antidepressant were followed for 13 to 16 months after the initial prescription. Outcome measures were rates of treatment termination before one month and subtherapeutic dosing, defined as having received no prescribed daily dosages at or above commonly cited thresholds. RESULTS: Among tricyclic-treated patients, psychiatrists' patients were significantly more likely than nonpsychiatrists' patients to continue in treatment for more than one month (72 percent versus 62 percent). Among patients taking tricyclics for at least three months, those with at least one prescription from a psychiatrist had a significantly higher rate of therapeutic dosing than those with all prescriptions from a nonpsychiatrist (70 percent versus 25 percent). For SSRI-treated patients, rates of termination and therapeutic dosing did not differ significantly by prescriber type. In multivariate equations that controlled for selected differences, effects of drug type and prescriber type were independent when persistence in treatment was analyzed, and interactive when subtherapeutic dosing was analyzed. CONCLUSIONS: Policy making about antidepressant pharmacotherapy should include assessments of the relationships between drug selection and patient outcome across a variety of clinical settings. PMID- 9735960 TI - Predictors and outcome of discharge against medical advice from the psychiatric units of a general hospital. AB - OBJECTIVE: The study examined predictors of discharge against medical advice (AMA) and outcomes of psychiatric patients with AMA discharges, as measured by poorer symptom ratings at discharge and higher rates of rehospitalization. METHODS: A total of 195 patients discharged AMA from general hospital psychiatric units were compared retrospectively with 2,230 regularly discharged patients. AMA status was defined as signing out against medical advice, being absent without leave, or being administratively discharged. All patients received standardized assessments within 24 hours of admission and at discharge. Demographic characteristics, psychiatric history, DSA-IV psychiatric and substance use diagnoses, and scores on an expanded 32-item version of the Psychiatric Symptom Assessment Scale were compared. RESULTS: The groups did not differ in primary psychiatric diagnoses. Patients discharged AMA were significantly less likely to be Caucasian or to be functionally impaired due to physical illness. They were more likely to live alone, have a substance use diagnosis, use more psychoactive substances, and have more previous hospitalizations. Patients discharged AMA had significantly shorter lengths of stay, higher rehospitalization rates, and more severe symptoms at discharge, even when length of stay was taken into account. The differences between the groups in male gender and young age were better accounted for by a greater likelihood of substance abuse in these groups. CONCLUSIONS: The results suggest a profile of patients who may be discharged AMA. Such patients have worse outcomes and are more likely to be high utilizers of inpatient resources. Aggressive identification of patients likely to be discharged AMA and early discharge planning for appropriate outpatient treatment are recommended. PMID- 9735961 TI - Competence to consent to voluntary psychiatric hospitalization: a test of a standard proposed by APA. American Psychiatric Association. AB - OBJECTIVE: In the wake of the U.S. Supreme Court's 1990 decision in Zinermon v. Burch, renewed attention has been given to capacities patients must have to be considered competent to consent to voluntary hospitalization. An American Psychiatric Association (APA) task force suggested that strong policy interests support the establishment of a low threshold for competence in this situation. The study examined whether, as previous research suggested, patients would have difficulty meeting even this lenient standard. METHODS: One hundred voluntarily hospitalized psychiatric patients were read two brief paragraphs, one explaining the purposes of psychiatric hospitalization and and the other explaining policies for discharge. The paragraphs' readability measured about eighth-grade level. After each paragraph, participants were read two sets of questions, one testing recall of the presented information and the other testing recognition of the information in a true-false format. The scores of patients grouped by selected demographic and clinical variables were compared. RESULTS AND CONCLUSIONS: The vast majority of patients were able to comprehend the information that the APA task force suggested was relevant to their decision. However, a subgroup of patients who were initially admitted involuntarily had significantly poorer performance and may constitute a group who need special educational efforts focused on the consequences of voluntary admission. PMID- 9735962 TI - Guidelines for prescribing psychiatrists in consultative, collaborative, and supervisory relationships. AB - Most psychiatrists enter into a variety of professional relationships with other clinicians in which they prescribe medications or make recommendations about pharmacotherapy. This paper describes a set of guidelines for prescribing psychiatrists involved in consultation, collaboration, and supervision with other clinicians. The guidelines were developed by psychiatrists for the Harvard Risk Management Foundation. The terms consultation, collaboration, and supervision are defined, and the psychiatrist's roles and responsibilities in each type of arrangement are described. The guidelines limit consultation and collaboration to relationships with professionals who are licensed or credentialed. Based on the definitions, the paper describes a structure for working with other clinicians, which begins with a thorough assessment of the context and circumstances of the clinical situation. The guidelines strongly encourage structured communication among clinicians and with the patient and significant others, as well as clarification by clinicians of their respective responsibilities for treatment and follow-up. PMID- 9735963 TI - The role of the psychiatrist as program medical director. AB - OBJECTIVE: In a recently published survey of alumni of the Columbia University public psychiatry fellowship, respondents who were medical directors reported performing a greater variety of tasks and experiencing higher job satisfaction than those who were staff psychiatrists. Both medical directors and staff psychiatrists believed that job satisfaction was most dependent on clinical collaboration activities. Survey data were reanalyzed to determine whether there was a relationship between the frequency of tasks performed and overall job satisfaction, and whether the tasks that actually predicted overall job satisfaction were the same as those that respondents believed contributed to job satisfaction. METHODS: The survey was distributed to all public psychiatry fellows and alumni in active practice (N=89), and 72 forms (81 percent) were returned. The survey consisted of 16 self-administered items divided into three categories of job tasks: direct service, clinical collaboration, and administration. RESULTS AND CONCLUSIONS: Despite respondents' beliefs that clinical collaboration activities contributed most to job satisfaction, performance of administrative tasks was found to best correlate with overall job satisfaction. Furthermore, overall job satisfaction was related to the performance of administrative tasks and not to the job title of medical director alone. Most of the medical directors in the survey had program-level, rather than agency-level, responsibilities. The findings indicate that the role of program medical director can serve as a crucial next step for staff psychiatrists, offering the opportunity to perform administrative tasks, which, according to the results, improves job satisfaction in public-sector positions. PMID- 9735964 TI - A three-decade perspective on community and public psychiatry training in Oregon. AB - The public psychiatry training program at Oregon Health Sciences University, established in 1973, educates psychiatric residents to work in community mental health centers and state hospitals. The authors present a brief history of this program, which spans three decades, and describe recent developments in its operation, with special attention to financing, administrative structure, and educational elements. Several program graduates have chosen careers in public sector work. The program is founded on the principle that just as dollars should follow patients in health care systems, so should residents in training follow patients. Administrative and fiscal arrangements must be flexible to support this mobility. PMID- 9735965 TI - Factors associated with involuntary return to a psychiatric emergency service within 12 months. AB - OBJECTIVE: This study examined patient characteristics and other factors that contributed to the involuntary return of patients to a psychiatric emergency service within 12 months of an initial evaluation in the service. The findings were used to consider whether the pressure to limit duration of hospital stays under managed care contributed to the patients' return to the emergency service. METHODS: Structured observations of evaluations of 417 patients admitted to the psychiatric emergency service were completed at seven county general hospitals in California. Twelve months after the initial evaluation, mental health and criminal justice records were reviewed for evidence of the patients' return for emergency psychiatric evaluation at any of the seven hospitals. Factors associated with patients' return to the psychiatric emergency service were evaluated using multivariate modeling. RESULTS: Of the 417 patients initially evaluated, 121, or 29 percent, were involuntarily returned to the psychiatric emergency service within 12 months. The likelihood of involuntary return was increased by a psychotic diagnosis and indications of dangerousness at the initial evaluation. Having insurance also increased the likelihood of involuntary return. CONCLUSIONS: The patient's initial condition in the psychiatric emergency service was found to be the best predictor of involuntary return. Brief hospitalization--an average of six days--after the evaluation did not have a significant prophylactic effect, perhaps because the reduced length of inpatient stay in the managed care environment did not allow adequate resolution of the patient's clinical condition. PMID- 9735966 TI - An agency-based representative payee program and improved community tenure of persons with mental illness. AB - OBJECTIVE: Representative payee programs help severely mentally ill individuals manage money from their Social Security payments to cover expenses for necessities and to avoid homelessness and rehospitalization. This study examined a representative payee program operated by a community mental health center to determine the criteria used by clinicians and ease managers to refer clients to the program and to learn whether participation in the program was associated with reductions in hospitalization. METHODS: The retrospective study included 56 individuals with severe mental illness who were enrolled in the representative payee program at Community Counseling Centers of Chicago for one year and who also had received services from the agency for at least one year before enrollment. Criteria used to refer clients to the representative payee program were determined through chart reviews. Data on state hospitalizations before and after enrollment were available for the entire sample; additional data on Medicaid-funded private hospitalizations were available for a subset of 33 clients. RESULTS: The most common criteria for enrollment in the representative payee program were comorbid substance abuse or dependence (49 percent), a history of homelessness (33 percent), and frequent hospitalizations (32 percent). During the year of participation in the representative payee program, the mean number of days spent in state hospitals decreased markedly compared with the year before enrollment, from 68 days to seven days. A similar reduction was noted in the number of days spent in state and private hospitals, from 97 days to 15 days. CONCLUSIONS: Findings from this pre- and postintervention retrospective study are tentative in the absence of a more rigorous design. However, the results suggest that the representative payee program is quite effective in reducing hospital stays. PMID- 9735967 TI - Characteristics of persons with mental illness in a representative payee program. AB - This study compared the characteristics of 56 clients with severe mental illness in a community mental health agency's representative payeeship program with those of 54 clients who did not participate in the program. Based on data from a two year period, participants in the representative payee program were characterized by disability or financial distress, indicated by a diagnosis of schizophrenia, homelessness, lack of rent money, and lack of financial skills; long-term dependence on income from Social Security and services provided by the mental health system, evidenced by receipt of Supplemental Security Income and frequent hospitalizations; and lack of financial independence, as reflected by inability to earn income from employment and lack of financial support from family. PMID- 9735968 TI - Differential access to care for children with ADHD in special education programs. AB - Access to treatment for children with attention-deficit hyperactivity disorder (ADHD) was examined in the general health, specialty mental health, and informal care sectors. Special education students in a Florida school district were screened for ADHD, and high-risk children and their parents completed diagnostic and services assessment interviews. Female gender, minority status, and rural residence lowered the probability of ADHD service use in the general health sector. Use of services in the mental health and informal sectors was predicted by a child's need for services. Further study is needed to identify barriers to service use at the parental or gatekeeper level for this common disorder among children. PMID- 9735969 TI - Reliability and validity of the interview and self-report versions of the BASIS 32. AB - The psychometric properties of the interview and self-report versions of the BASIS-32 were compared. A total of 120 severely mentally ill adults enrolled in psychosocial rehabilitation were randomly assigned to either a self-report or an interview condition. The BASIS-32 had good internal consistency and test-retest reliability on most subscales; coefficients were higher in the self-report condition. Only the interview version of the psychosis subscale had unacceptable internal consistency. Validity correlations were generally good for the symptom subscales but disappointing for the functional domains. The subscale scores did not discriminate between diagnostic subgroups. PMID- 9735970 TI - Compliance with depot antipsychotic medication by patients attending outpatient clinics. AB - Outpatients with schizophrenia often have problems complying with a regimen of oral antipsychotic medications. Use of depot medications improves compliance, but patients' geographic location and access to transportation may affect compliance with a depot regimen. The authors used retrospective chart review to examine whether patients' geographic location and sociodemographic characteristics and characteristics of their medication use were related to compliance with a depot regimen. The subjects were 75 patients attending an urban clinic and 23 patients attending a rural clinic in New York State. Median rates of compliance were 94.7 percent for the urban patients and 96.4 percent for the rural patients, not a significant difference. The only characteristic associated with a decreased compliance rate was a history of substance abuse. PMID- 9735971 TI - Utah's capitation plan and the process of care. PMID- 9735972 TI - Depot neuroleptics in Canada. PMID- 9735973 TI - Average cost of newer psychotropic, antidepressant drugs twice as high in U.S. as in Europe, study finds. PMID- 9735974 TI - Study shows employers win 92 percent of court cases involving discrimination against disabled employees. PMID- 9735975 TI - Pleural effusions: patterns on ventilation-perfusion lung scans. AB - Perfusion and ventilation abnormalities created by pleural effusions can interfere with the interpretation of the lung scan. This retrospective study identified and evaluated the specificity of scintigraphic patterns for pleural effusion. Ninety-two ventilation-perfusion lung scans were analyzed for the following signs of pleural effusion: presence of fissures, straightened or concave lung borders, costophrenic angle blunting, and attenuation of lung activity by interposed fluid. The findings later were correlated with chest radiographs. Of 25 pleural effusions detected by chest radiography, scintigraphy predicted 14 (specificity, 86%). In all of these cases, there was agreement with the chest radiograph (specificity, 100%). The fissure sign and the straight concave border sign were equally reliable for the prediction of pleural fluid. Costophrenic angle blunting was never seen as the sole indicator of pleural fluid, and attenuation was seen alone in only one case. Another finding observed during this evaluation was absent ventilation at the lung base with preserved perfusion. Scintigraphic patterns may not be reliable in obstructive lung disorders and diseases with altered lung compliance. The recognition of scintigraphic patterns of pleural effusions on ventilation-perfusion scans can improve the clinical value of lung scintigraphy by reducing the number of indeterminate readings. PMID- 9735976 TI - The 24-hour Tl-201 image in dual isotope myocardial perfusion scintigraphy: clinical utility and prognostic significance. AB - This study assessed the effect on clinical decision making and the possible prognostic significance of the 24-hour Tl-201 image in patients undergoing Tl-201 MIBI dual-isotope myocardial scintigraphy. The records of all patients who underwent 24-hour Tl-201 imaging as part of their myocardial perfusion study from 1994 to 1996 were reviewed. Follow-up evaluations were obtained from the referring physician or by direct patient contact. Fifty-six patients underwent a total of 57 studies; four patients were lost to follow-up. Of the 53 studies evaluated, 29 showed no change between the standard rest images and the 24-hour images; these patients were reported to have myocardial scar. Of these 29 patients, 25 were treated medically without further evaluation; 24 of these 25 patients remained stable. Four of the 29 patients had further evaluation; 2 patients had coronary artery bypass graft, 1 had a stent placed, and 1 remained stable. Twenty-four patients showed definite improvement or normalization of their study results by 24 hours; they were reported as ischemic. Of these 24 patients, 11 were treated medically without further evaluation; 9 remained stable, whereas 2 had adverse events. The remaining 13 patients required further evaluation; 4 remained stable, whereas 9 had adverse events (4 = increasing angina; 1 = stent; 1 = rotoblator; 2 = percutaneous transluminal coronary angioplasty; 1 = death). Twenty-four-hour imaging contributes to clinical decision making and may identify a subset of patients at risk for subsequent complications. PMID- 9735977 TI - Perivalvular abscess complicating infective endocarditis: complementary role of echocardiography and indium-111-labeled leukocytes. AB - A 49-year-old black man with hypertension-induced chronic renal failure requiring hemodialysis and a history of arteriovenous access graft infection was admitted with Staphylococcus aureus sepsis, dyspnea, and peri-incisional erythema over his arteriovenous graft fistula. Results of a transthoracic echo demonstrated aortic sclerosis and concentric left ventricular hypertrophy. Results of a whole-body In 111 white cell (WBC) scan were negative over the arteriovenous graft site; however, an intense abnormal focus of labeled WBCs was evident to the left of the sternum. A subsequent transesophageal echocardiogram showed a mixed cystic-solid calcified mass adjacent the left aortic cusp. Surgery confirmed a perivalvular abscess. As a whole-body imaging modality, the In-111 WBC scintigram indicated the true location of the infectious process responsible for the patient's sepsis. The combination of echocardiography and radiolabeled WBC imaging increases sensitivity for detection of endocarditis/perivalvular abscess. Radiolabeled WBC imaging is more efficacious for monitoring therapy because the echocardiogram often does not change with treatment of endocarditis/perivalvular abscess. PMID- 9735978 TI - I-131 uptake in the breast for thyroid cancer surveillance with biopsy-proven benign tissue. AB - A 37-year-old woman was seen for recurrent papillary carcinoma of the thyroid after thyroidectomy. After repeated surgery and I-131 therapy, follow-up I-131 scanning and thyroglobulin levels were negative. Subsequent I-131 surveillance, however, demonstrated bilateral breast uptake. A biopsy taken of this area in the right breast proved that the increased uptake was secondary to benign disease. PMID- 9735979 TI - Reverse discordant scintigraphy in diffuse goiter. AB - Both Tc-99m pertechnetate and radioactive iodine (I-123 NaI or I-131 NaI) are useful in thyroid scintigraphy. These radiopharmaceuticals yield similar functional information in most patients. Occasionally, however, discordant results have been reported in the literature (warm or hot on the pertechnetate image and cold on the radioiodide image). Most of these reports have concerned the solitary thyroid nodule. A case is presented here with diffusely decreased Tc 99m pertechnetate uptake and normal I-131 NaI uptake in a patient with a diffuse goiter and subclinical hyperthyroidism, so-called reverse discordant behavior. PMID- 9735980 TI - Hepatobiliary scintigraphy in an immunosuppressed patient with hepatocellular dysfunction and acute cholecystitis: diagnostic dilemmas posed by delayed gallbladder visualization. AB - A case of acute cholecystitis in an immunosuppressed patient with hepatocellular dysfunction is reported. The diagnostic dilemmas posed by the lack of specific sonographic findings, the possibility of acute gallbladder disease without early cystic duct obstruction, and the absence of clear guidelines for the interpretation of delayed appearance of the gallbladder on hepatobiliary scintigraphy in this subgroup of patients are discussed. PMID- 9735981 TI - Tc-99m MIBI thoracic SPECT for the detection of intrathoracic tumor masses. AB - Thirty-one men (age range, 46-76 years; mean age, 64.8 years) with intrathoracic masses suggesting possible malignancy on the basis of chest radiography or CT underwent preoperative Tc-99m MIBI SPECT examinations. Diagnosis was confirmed on pathologic examinations of samples obtained either at thoracotomy, esophagectomy, or by biopsy. Twenty-five patients had primary lung cancer, including squamous cell carcinoma, large cell carcinoma, adenocarcinoma, and small cell carcinoma. Two patients had lymphomas with spread to the mediastinum, and three patients had extrathoracic primary cancers (one squamous cell carcinoma of esophagus, one squamous cell carcinoma originating from a head and neck tumor, and one metastatic mediastinal leiomyosarcoma). One patient with a tuberculoma had negative results of the Tc-99m MIBI examination. Tc-99m MIBI had a 86.7% sensitivity rate, a 0% false-positive rate, and a 100% positive predictive value to detect malignant intrathoracic masses. There was a 13% false-negative rate, however, suggesting that MIBI-SPECT may underdiagnose malignant lesions. SPECT findings of these 31 patients can be classified as 1) mass with increased uptake, n = 23; 2) ring-like appearance of increased uptake, n = 3; 3) mass with absent uptake, n = 4; and 4) photon-deficient mass, n = 1. Absent uptake in patients with mass lesions could be explained by necrosis of the lesion (caseation necrosis or massive tumor necrosis with or without bleeding). Most malignant intrathoracic masses are Tc-99m MIBI avid and may be detected with a high degree of sensitivity and with an excellent positive predictive value. A positive MIBI scan may help in the clinical diagnosis of malignancy. The use of Tc-99m MIBI could serve not only as a tumor imaging agent, but also may be used to determine the extent of spread and potentially the chemotherapeutic responsiveness of a tumor. PMID- 9735982 TI - Follow-up of improvement in regional cerebral blood flow and mental status in Alzheimer's disease: a case report. AB - Cerebral SPECT imaging has the potential to make important contributions in the follow-up care of patients with Alzheimer's disease. An unusual case of a patient who showed a follow-up cerebral blood flow pattern different from that routinely seen in Alzheimer's disease patients is reported here. Qualitative and voxel based objective evaluation of follow-up scans revealed improvement in parietotemporal deficits that had been observed on a baseline study. This change was observed without significant further deterioration in ratings by dementia batteries. This case shows that a baseline regional cerebral blood flow study might be necessary for reference and comparison in the proper follow-up care of Alzheimer's disease patients. PMID- 9735983 TI - F-18 FDG whole-body positron emission tomography scan in primary breast sarcoma. AB - Three patients with primary breast sarcoma showed intense F-18 FDG breast uptake on the whole-body scan. In two patients the uptake was irregular and associated with cold foci that corresponded to hypodense lesions noted on the chest CT; these represented areas of pathologically demonstrated tumor necrosis. There was also intense FDG uptake in pulmonary, axillary, and supraclavicular lymph node metastases. All lesions were confirmed by CT scan of the chest. Thus F-18 FDG positron emission tomographic scanning accurately staged the tumors in these two patients, and it documented local recurrence in the third patient. Histopathologic examination showed evidence of a high-grade sarcoma, a primary rhabdomyosarcoma, and a malignant cystosarcoma phyllodes of the breast. Similar to breast carcinoma, F-18 FDG whole-body positron emission tomographic imaging could be useful in diagnosing and staging primary breast sarcomas. PMID- 9735985 TI - Asymptomatic periostitis of the navicular and fifth metatarsal bones in a high jumper. PMID- 9735984 TI - Iodine-123 BMIPP and Ga-67 scintigraphy in liposarcoma. AB - A patient with recurrent retroperitoneal liposarcoma had multiple suspected metastases. I-123 BMIPP imaging showed areas of increased uptake due to accumulation in the myxoid components of the liposarcoma. Ga-67 showed accumulation in the undifferentiated components. The well-differentiated components showed little accumulation of either I-123 BMIPP or Ga-67. Differences in the accumulation of these radionuclides may reflect differences in cell densities, fatty acid metabolism, and the degree of malignancy. I-123 BMIPP and Ga-67 scintigraphy may be useful in determining the prognosis of liposarcoma. PMID- 9735986 TI - Tc-99m MDP uptake in primary breast lymphoma. PMID- 9735987 TI - Incidental second primary in the breast detected by F-18 FDG positron emission tomography scan. PMID- 9735988 TI - The usefulness of Tc-99m sestamibi scintimammography in the diagnosis of squamous cell carcinoma of the breast. PMID- 9735989 TI - Primary osteogenic sarcoma of the breast demonstrated by Tc-99m MDP scintigraphy. PMID- 9735990 TI - Tc-99m MDP uptake in stage D adenocarcinoma of the colon. PMID- 9735991 TI - Hiatus hernia: a potential cause of false-positive iodine-131 scan in thyroid carcinoma. PMID- 9735993 TI - Spontaneous regression of hepatic pseudotumor demonstrated by Ga-67 imaging. PMID- 9735992 TI - Giant pseudoaneurysm of the left ventricle diagnosed by radionuclide ventriculography. PMID- 9735994 TI - Eventration of the diaphragm on radionuclide ventriculography. PMID- 9735996 TI - Abnormal cortical metabolism in acute disseminated encephalomyelitis. PMID- 9735995 TI - Increased Tc-99m MIBI uptake in atelectatic lung: a pitfall in scintigraphic evaluation of central bronchogenic carcinoma. PMID- 9735997 TI - Ga-67 activity in the interposed free jejunum mimicking malignant lesions. PMID- 9735998 TI - Tc-99m RBC SPECT demonstrating vertebral hemangioma. PMID- 9735999 TI - Tc-99m HMPAO brain perfusion SPECT images in a patient with portal-systemic encephalopathy. PMID- 9736000 TI - Tc-99m sestamibi imaging as an indicator of P-glycoprotein expression in metastatic pheochromocytoma. PMID- 9736001 TI - Current readings in nuclear medicine. PMID- 9736002 TI - Lifetime prevalence of DSM-III-R psychiatric disorders among urban and rural Mexican Americans in California. AB - BACKGROUND: The Mexican American Prevalence and Services Survey presents lifetime prevalence rates for 12 DSM-III-R psychiatric disorders in a sample of 3012 adults of Mexican origin by place of residence and nativity, and compares these results with those of population surveys conducted in the United States and Mexico. METHODS: The stratified random sample included non-institutionalized persons aged 18 to 59 years of Mexican origin, who were residents of Fresno County, California. Psychiatric disorders were assessed using a modified version of the World Health Organization Composite International Diagnostic Interview in face-to-face interviews. RESULTS: Mexican immigrants had lifetime rates similar to those of Mexican citizens, while rates for Mexican Americans were similar to those of the national population of the United States. This difference is attributable to a prevalence rate for any disorder among immigrants of 24.9%, compared with 48.1% among US-born respondents. A higher prevalence for any disorder was reported in urban (35.7%) compared with town (32.1%) or rural (29.8%) areas. Multivariate analyses showed an adjusted effect of country of birth, but not of urban residence. CONCLUSIONS: Despite very low education and income levels, Mexican Americans had lower rates of lifetime psychiatric disorders compared with rates reported for the US population by the National Comorbidity Survey. Psychiatric morbidity among Mexican Americans is primarily influenced by cultural variance rather than socioeconomic status or urban vs rural residence. PMID- 9736003 TI - Immigration and mental health: why are immigrants better off? PMID- 9736004 TI - Changing patterns of psychiatric inpatient care in the United States, 1988-1994. AB - Using data from the National Hospital Discharge Survey and the Inventory of Mental Health Organizations, this article examines national trends in psychiatric inpatient care from 1988 to 1994 in general hospitals and mental hospitals. We find that discharges with a primary diagnosis of mental illness in general hospitals increased from 1.4 to 1.9 million during this period. The total increase of 1.2 million days of care in general hospitals was small relative to the reduction of 12.5 million inpatient days in mental hospitals. General hospital discharges increased most in private nonprofit hospitals and declined substantially in public hospitals. Length of stay has fallen most substantially in private nonprofit hospitals. Public programs have increasingly replaced private insurance as the major source of payment. These observations suggest that psychiatric inpatient care in general hospitals can be characterized as a process in which patients who would have been clients of public mental hospitals in a prior period replace privately insured patients who, under managed care, are largely treated in community settings. Private nonprofit general hospitals increasingly treat publicly financed patients with more severe illnesses. PMID- 9736005 TI - Brain dopamine transporter messenger RNA and binding sites in cocaine users: a postmortem study. AB - BACKGROUND: Results of recent radioligand binding experiments suggest that chronic cocaine exposure increases dopamine transporter (DAT) synthesis throughout the striatum of humans. However, detection of cocaine binding site increases in animals and humans has varied depending on the radioligand used. The present experiment tested the hypothesis in cocaine-using humans that synthesis of midbrain DAT messenger RNA increases parallel with increased striatal DAT binding sites. METHODS: Striatal and midbrain samples were collected during autopsy examination from human cocaine users (n = 34) and from age-, sex-, and race-matched control subjects (n = 36). Levels of DAT messenger RNA were quantified in the medial and lateral midbrain regions using in situ hybridization, and striatal DAT binding sites were assessed by quantitative autoradiography using the DAT-specific radioligand [3H]WIN 35428. RESULTS: Striatal DAT binding sites were markedly increased in cocaine users, but, paradoxically, medial DAT messenger RNA levels were decreased. CONCLUSION: Cocaine exposure has a marked effect on DAT function, but the mechanisms involved may be complex. PMID- 9736006 TI - Lifetime panic-depression comorbidity in the National Comorbidity Survey. AB - BACKGROUND: The National Comorbidity Survey is a nationally representative survey of the prevalences and correlates of DSM-III-R disorders in the US household population. METHODS: Retrospective age-at-onset reports were used to study predictive relationships between lifetime panic and depression. RESULTS: Strong associations were found between the lifetime prevalences of panic and major depressive episodes (odds ratios: for panic attacks with depression, 6.2; for panic disorder with depression, 6.8). These associations were not significantly influenced by the inclusion or exclusion of respondents with mania. Temporally primary depression predicted a first onset of subsequent panic attacks but not of panic disorder. Temporally primary panic attacks, with or without panic disorder and whether or not the panic was persistent, predicted a first onset of subsequent major depression. The associations between panic attack and depression were attenuated in models that controlled for prior traumatic life experiences and histories of other DSM-III-R disorders. CONCLUSIONS: Lifetime panic depression comorbidity characterizes most community respondents with panic disorder and a substantial few of those with major depression. The absence of a dose-response relationship suggests that primary panic attack is a marker, rather than a causal risk factor, of subsequent depression. Primary depression, in comparison, appears to be a genuine risk factor for secondary panic attacks. That primary depression predicts panic attacks but not panic disorder suggests that secondary panic is a severity marker of depression rather than a comorbid condition. These results are far from definitive because they are based on retrospective reports, lay-administered diagnostic interviews, and only 1 survey. However, they raise important questions that could lead to a fundamental rethinking of panic-depression comorbidity if they are replicated in future epidemiological and clinical studies. PMID- 9736007 TI - The relationships between age, sex, and the incidence of dementia and Alzheimer disease: a meta-analysis. AB - BACKGROUND: Prevalence studies on dementia and Alzheimer disease (AD) have reported a positive association with age. However, the trend of the association in the oldest-old categories has been the subject of discussion. The relationship between sex and AD has been inconsistent with these studies. Prevalence rates are influenced by the survival and disease incidence. Incidence rates provide a better measure of disease risk. METHODS: English-language articles identified through a MEDLINE search on "incidence dementia" and "incidence Alzheimer's disease" were examined and references from identified articles were reviewed. Population-based studies using personal interviews, standard clinical diagnosis criteria (DSM-III for dementia, National Institute of Neurological and Communicative Disorders and Stroke-Alzheimer's Disease and Related Disorder Association for AD) and reporting age-specific incidence rates were included in the meta-analysis. Data from the selected studies were extracted and verified. Mixed-effect models were used in the meta-analysis to accommodate the heterogeneity of the studies. RESULTS: Incident dementia and AD are associated with a significant quadratic age effect indicating that the increase in incidence rates slows down with the increase in age, although there is no sign of a decline in the incidence rates themselves. The odds ratios for women to develop incidence of dementia and AD relative to men are 1.18 (95% confidence interval, 0.95-1.46) and 1.56 (95% confidence interval, 1.16-2.10), respectively. CONCLUSIONS: The acceleration of incidence rates for AD and dementia slows down with the increase in age, although we find no evidence of a rate decline. Women are at higher risk of developing AD than men. PMID- 9736008 TI - Prevention of recurrent depression with cognitive behavioral therapy: preliminary findings. AB - BACKGROUND: Cognitive behavioral treatment (CBT) of residual symptoms after successful pharmacotherapy yielded a substantially lower relapse rate than did clinical management in patients with primary major depressive disorders. The aim of this study was to test the effectiveness of this approach in patients with recurrent depression (> or = 3 episodes of depression). METHODS: Forty patients with recurrent major depression who had been successfully treated with antidepressant drugs were randomly assigned to either CBT of residual symptoms (supplemented by lifestyle modification and well-being therapy) or clinical management. In both groups, during the 20-week experiment, antidepressant drug administration was tapered and discontinued. Residual symptoms were measured with a modified version of the Paykel Clinical Interview for Depression. Two-year follow-up was undertaken, during which no antidepressant drugs were used unless a relapse ensued. RESULTS: The CBT group had a significantly lower level of residual symptoms after discontinuation of drug therapy compared with the clinical management group. At 2-year follow-up, CBT also resulted in a lower relapse rate (25%) than did clinical management (80%). This difference attained statistical significance by survival analysis. CONCLUSIONS: These results challenge the assumption that long-term drug treatment is the only tool to prevent relapse in patients with recurrent depression. Although maintenance pharmacotherapy seems to be necessary in some patients, CBT offers a viable alternative for other patients. Amelioration of residual symptoms may reduce the risk of relapse in depressed patients by affecting the progression of residual symptoms to prodromes of relapse. PMID- 9736009 TI - Distinct contributions of conduct and oppositional defiant symptoms to adult antisocial behavior: evidence from an adoption study. AB - BACKGROUND: We conducted an exploratory multivariate analysis of juvenile behavior symptoms in an adoption data set. One goal was to see if a few DSM interpretable symptom dimensions economically captured information within the data. A second goal was to study the relationships between any such dimensions, biological and environmental background, and eventual adult antisocial behavior. METHODS: The data originated from a retrospective adoption study. Probands with a biological background for parental antisocial personality or alcoholism were heavily oversampled. Symptoms were ascertained by proband and adoptive parent interview. We performed, by gender, orthogonal rotated principal component analyses of juvenile behavior disturbance symptoms (females, n = 87; males, n = 88). We used structural equation modeling to examine the relationships hypothesized above. RESULTS: For both genders, an oppositional defiant disorder (ODD) component and at least 1 conduct component emerged. Regardless of the conduct component scores, the ODD components were significant predictors of adult antisocial behavior. For males, the ODD component was predicted by an antisocial biological background, but not by scores on the Adverse Adoptive Environment Scale. The conduct components were predicted by adoptive environment alone. For females, biological background or biological-environmental interactions predicted each of the components. CONCLUSIONS: There has been little previous distinction between conduct disorder and ODD in studies of genetic and environmental influences on juvenile behavior. The study suggests that adolescent ODD symptoms may be a distinct antecedent of adult antisocial personality. In males, adolescent ODD symptoms may represent early expression of genetic sociopathic personality traits. PMID- 9736010 TI - Smooth pursuit eye movements to extraretinal motion signals: deficits in relatives of patients with schizophrenia. AB - BACKGROUND: Although mounting evidence supports the idea that smooth pursuit abnormality marks the genetic liability to schizophrenia, the precise ocular motor mechanism underlying the abnormality remains unknown. Based on recent findings in schizophrenia, we hypothesize that subtle deficits in the ability to hold online and/or use extraretinal motion information underlie the pursuit abnormality in vulnerable individuals. METHODS: The hypothesis was tested in 69 first-degree, biological relatives of probands with schizophrenia; 26 relatives had schizophrenia spectrum personalities (SSP). Subjects recruited from the community (n=71; 29 with SSP), without a known family history of psychosis, constituted the comparison groups. The traditional smooth pursuit gain measure, which is a ratio of smooth pursuit eye velocity in response to both retinal and extraretinal motion signals and the target velocity, was obtained. In addition, newly developed measures of predictive smooth pursuit (ie, in the presence of only extraretinal motion signals) were obtained. The latter measures were evaluated after the current retinal motion signals were made unavailable by briefly making the target invisible. RESULTS: Relatives, particularly those with SSP, showed significantly poorer predictive pursuit response to extraretinal motion signals (F(2,136)=6.51, P<.005), compared with the community subjects. However, the traditional smooth pursuit gain in response to both retinal and extraretinal motion signals was not different between groups. CONCLUSIONS: These results suggest that relatives of patients with schizophrenia, particularly those with SSP, have specific deficits in predictive pursuit based on only extraretinal motion signals. Normal smooth pursuit gain in response to both retinal and extraretinal motion signals is likely due to compensation based on retinal motion information. The latter suggests normal retinal motion processing and smooth pursuit motor output. PMID- 9736011 TI - Antisaccades and smooth pursuit eye tracking and schizotypy. AB - BACKGROUND: Eye tracking deficits are one of a few widely validated behavioral markers of risk for schizophrenia. Recently, it has been proposed that antisaccade performance may also constitute a marker of schizophrenia risk. This study investigated whether eye tracking and antisaccade deficits could be found in another population with putative liability to schizophrenia-nonclinical subjects with elevated scores on a psychometric index of perceptual aberrations. METHODS: Subjects were 55 university students who received either high or normal scores on the Perceptual Aberration Scale, a measure of schizotypy indexing body image and perceptual distortions. Subjects completed a smooth pursuit eye tracking task and an antisaccade task. Eye movements were monitored using an infrared limbus tracker. RESULTS: Subjects with high Perceptual Aberration Scale scores (putative "schizotypes") had lower pursuit quality and a lower percentage correct on the antisaccade task than the controls. The 2 groups did not differ in antisaccade or error latencies. The increase in antisaccade errors in the schizotypes was accounted for almost entirely by an increase in perseverative errors, but virtually no difference between groups on random errors. Antisaccade performance was significantly related to pursuit quality. CONCLUSIONS: Subjects with elevated Perceptual Aberration Scale scores have performance deficits on oculomotor tasks that have been linked to latent liability to schizophrenia, namely, smooth pursuit and antisaccade performance. The antisaccade errors in the schizotype group were primarily perseverations, a behavioral pattern often associated with frontal lobe dysfunction and observed in the performance of schizophrenic patients. PMID- 9736012 TI - Distinguishing apathy syndromes from vascular depression. PMID- 9736013 TI - Does clozapine treatment cause brain disease? PMID- 9736014 TI - Ocular neovascularization: clarifying complex interactions. PMID- 9736015 TI - Multiple system atrophy: clues from inclusions. PMID- 9736016 TI - The fate of T cells in the brain: veni, vidi, vici and veni, mori. PMID- 9736017 TI - Down-regulation of p27 is associated with development of colorectal adenocarcinoma metastases. AB - The cyclin-dependent kinase inhibitor p27 is a negative regulator of the cell cycle and a potential tumor suppressor gene. Because we had previously demonstrated that loss of p27 protein is associated with aggressive behavior in colorectal adenocarcinomas, we used immunohistochemistry and in situ hybridization to evaluate the potential role of alterations in p27 expression in primary and metastatic colorectal adenocarcinomas. Parallel immunostaining was performed for Ki-67 and p53. We evaluated 13 cases of metachronous and 23 cases of synchronous primary and metastatic colorectal tumor pairs. In the synchronous subgroup (Stage IV tumors), 57% of the primary tumor and metastases pairs did not express p27 protein and the remainder were low expressors. In the metachronous subgroup, 54% of the primary tumors were low expressors and the remainder high expressors of p27 protein. There was a significant reduction in the expression of p27 in the metachronous metastases (mean positive cells: 14.5%) when compared to the corresponding primary tumors (mean positive cells: 41.8%), P = 0.0023. All the primary and metastatic tumors in the metachronous subgroup showed high levels of p27 mRNA expression. There was no association between loss of p27 and either Ki-67 count or p53 expression. Because p27 is known to be up-regulated when epithelial cells are grown in suspension, the down-regulation of p27 in circulating tumor cells may confer the ability to grow in an environment of altered extracellular matrix or intercellular adhesion properties, two situations which may facilitate metastases. PMID- 9736018 TI - Trisomy 3 is not a common feature in malignant lymphomas of mucosa-associated lymphoid tissue type. AB - The genetic background of extranodal marginal zone B-cell non-Hodgkin's lymphoma (NHL) of mucosa-associated lymphoid tissue (MALT) type is poorly understood. In contrast to most entities of primary nodal lymphomas, few cytogenetic data are available, and gene rearrangements frequently encountered in and highly characteristic of certain entities of systemic NHL are absent in this type of lymphoma. Recently, it was suggested that MALT-type NHLs are associated with certain numerical chromosome aberrations and especially with trisomy 3. We performed an extensive study using a sensitive double (bicolor) fluorescence in situ hybridization technique for the analysis of trisomies for chromosomes 3, 7, 12, and 18 in 60 samples of low-grade and 45 high-grade MALT-type tumors. In the low-grade cases, trisomy 3 was found in a frequency of only 20%. High-grade lymphomas of MALT type were associated with trisomies 3, 7, 12, and 18 in 36, 20, 18, and 13% of the cases, respectively. Whereas no difference was encountered for trisomy 3 in primary and secondary/simultaneous high-grade lymphomas, +7 and +12 were associated with primary lymphomas, and a +18 was predominantly found in secondary/simultaneous high-grade NHL. These results challenge earlier reports describing a high frequency of +3 in low-grade MALT-type NHL and indicate a possibly different genetic evolution pattern of primary and secondary/simultaneous high-grade lymphomas of primary mucosal origin. PMID- 9736020 TI - Porphyrin loading of lipofuscin granules in inflamed striated muscle. AB - To further the understanding of oxidative effects on inflammation injury to muscle fiber structure, fluorescent imaging analysis of human striated muscle tissues from a variety of inflammatory or postinflammatory etiologies was undertaken in a search for accumulated coproporphyrin, a red autofluorescent byproduct of heme biosynthesis that would theoretically be formed under oxidative insult. Using a differential excitation method of in situ analysis, porphyrin autofluorescence was detected in intact fibers within the context of the yellow autofluorescent subsarcolemmal lipofuscin granules. Relative measurements of porphyrin concentration in the granules from different patients indicated that the acute/subacute inflammatory specimens grouped significantly higher than the more chronic inflammatory and nonpathological specimens. Myoglobin was also found to be associated with the granules. Myoglobin heme iron could potentially serve as a Fenton reagent for the intracellular generation of hydroxyl radicals, which are responsible for the oxidation of the porphyrinogens. High-performance liquid chromatography analysis of extracted dense particles revealed coproporphyrin as the sole porphyrin present. The observation of coproporphyrin within lipofuscin granules, previously unreported, suggests that lipofuscin accumulation in striated muscle may begin under conditions of acute oxidative stress, as marked by the oxidation of extramitochondrial porphyrinogens that are immediately incorporated into the granules. PMID- 9736019 TI - Nonneural nuclear inclusions of androgen receptor protein in spinal and bulbar muscular atrophy. AB - Spinal and bulbar muscular atrophy is an X-linked motor neuronopathy caused by the expansion of an unstable CAG repeat in the coding region of the androgen receptor (AR) gene. Nuclear inclusions of the mutant AR protein have been shown to occur in the spinal motor neurons of spinal and bulbar muscular atrophy (Li M, Kobayashi Y, Merry D, Tanaka F, Doyu M, Hashizume Y, Fischbeck KH, Sobue G: Nuclear inclusions in spinal and bulbar muscular atrophy. Ann Neurol 1998 (in press)). In this study, we demonstrate the tissue-specific distribution, immunochemical features, and fine structure of nuclear inclusions of spinal and bulbar muscular atrophy. Nuclear inclusions were observed in affected spinal and brainstem motor neurons, but not in other, nonaffected neural tissues. Similar nuclear inclusions occurred in nonneural tissues including scrotal skin, dermis, kidney, heart, and testis, but not in the spleen, liver, and muscle. These inclusions had similar epitope features detectable by antibodies that recognize a small portion of the N-terminus of the AR protein only, and they were ubiquitinated. Electron microscopic immunohistochemistry showed dense aggregates of AR-positive granular material without limiting membrane, both in the neural and nonneural inclusions. These findings indicate that nuclear inclusions of AR protein are present in selected nonneural tissues as well as in neurons that degenerate in spinal and bulbar muscular atrophy, suggesting that a common mechanism underlies in the formation of neural and nonneural nuclear inclusions. PMID- 9736021 TI - Adenomatous polyposis coli gene mutation alters proliferation through its beta catenin-regulatory function in aggressive fibromatosis (desmoid tumor). AB - Aggressive fibromatosis is a monoclonal proliferation of spindle (fibroblast like) cells. A subset of lesions contain somatic truncating adenomatous polyposis coli (APC) gene mutations, and all of the lesions contain an elevated beta catenin protein level. A major function of APC is to regulate beta-catenin protein level. Beta-catenin has a dual function in the cell: it is a member of the adherens junction, and it binds transcription factors in the tcf-lef family, transactivating transcription. Cell cultures from aggressive fibromatoses containing an APC mutation were studied. Transient transfection of the full length APC gene caused decreased proliferation and beta-catenin protein level in these cultures. To determine whether beta-catenin protein level was responsible for the change in proliferation rate, stable transfections of deltaN89beta catenin (a stabilized form that is not degraded by APC, but retains all other functions) were achieved in half of the cultures derived from each tumor, whereas the other half were transfected with an empty vector. Transfection of the full length APC gene in cultures that were stably transfected with deltaN89beta catenin did not result in a change in proliferation. The type I promotor of p56lck contains an HMG consensus region, to which members of the tcf-lef family can bind. p56lck was expressed in cultures not transfected with the full-length APC gene and in cultures transfected with the full-length APC gene and deltaN89beta-catenin, but not in cultures transfected with only the full-length APC gene. These data show that APC truncating mutations give aggressive fibromatosis cells a proliferative advantage through beta-catenin and suggest that beta-catenin acts to transactivate transcription. PMID- 9736022 TI - T-cell apoptosis in inflammatory brain lesions: destruction of T cells does not depend on antigen recognition. AB - Elimination of inflammatory T cells by apoptosis appears to play an important role in the down-regulation of inflammation in the central nervous system. Here we report that apoptosis of T lymphocytes occurs to a similar extent in different models of autoimmune encephalomyelitis. Apoptosis is restricted to cells located in the neuroectodermal parenchyma, thereby leaving T cells present in the brain's connective tissue compartments unharmed. Death of T cells in the parenchyma does not depend on antigen presentation by resident microglial cells or astrocytes. Adoptive transfer experiments with T lymphocytes carrying a specific genetic marker revealed that in the central nervous system these cells are destroyed regardless of their antigen specificity or state of activation. Although many of both antigen-dependent and -independent mechanisms in the induction of T-cell apoptosis may act simultaneously, our results suggest that the nervous system harbors a specific, currently undefined, mechanism that effectively eliminates infiltrating T lymphocytes. PMID- 9736024 TI - Myelin degeneration in multiple system atrophy detected by unique antibodies. AB - A rabbit antiserum (anti-EP), induced against a synthetic peptide corresponding to residues 68 to 86 of guinea pig myelin basic protein, powerfully immunostained abnormal-appearing oligodendrocytic processes and cell bodies in demyelinating areas associated with multiple system atrophy (MSA). However, as we reported previously, the antiserum, which is highly specific for the sequence QDENPVV corresponding to human myelin basic protein residues 82 to 88, failed to recognize any structures in normal human brain. QD-9, a mouse monoclonal antibody raised against human myelin basic protein residues 69 to 88, which also recognizes specifically the epitope QDENPVV, gave the same results as did anti EP. The unusual epitope recognized by anti-EP/QD-9 antibodies appears to be accessible in areas of myelin degeneration, and the antibodies have been shown to detect such areas in multiple sclerosis and infarcted brains. These antibodies detect myelin degeneration more widely than previous conventional methods. The present study emphasizes the importance of myelin degeneration in the pathogenesis of multiple system atrophy. PMID- 9736025 TI - Heterogeneity of dendritic cells in human superficial lymph node: in vitro maturation of immature dendritic cells into mature or activated interdigitating reticulum cells. AB - A two-color immunofluorescent analysis indicated that dendritic cells (DCs) in the human axillar lymph nodes (ie, lymph nodal DCs (LnDCs)) can be classified into three subsets. The first subset consists of CD1a+/CD86(- or dim)/CD83(- or dim) nondendriform DCs found mainly in lymph sinuses, the second is of CD1a /CD86+/CD83+ dendriform DCs scattered in normal T zones, and the third is of large CD1a(bright)/CD86+/CD83+ dendriform DCs occasionally found in hyperplastic T zones. A three-color flow cytometric analysis, immunoperoxidase staining, and electron microscopic observation indicated that the majority of LnDCs corresponded to the first subset, which showed distinctive characteristics of DCs but did not fulfill the ultrastructural criteria for interdigitating reticulum cells (IDCs) and did not contain Birbeck granules. When LnDCs were cultured for 7 days, they became large CD1a(dim)/CD86+/CD83+ dendriform cells, which formed large complexes with many T cells and exhibited distinctive ultrastructural features of interdigitating reticulum cells. LnDCs cultured in the presence of granulocyte/macrophage colony-stimulating factor became markedly larger CD1a(bright)/CD86+/CD83+ dendriform cells forming large complexes with numerous T cells. These findings suggest that cells of the first subset represent immature LnDCs just migrating from epidermis, those of the second subset represent interdigitating reticulum cells, and those of the third subset represent interdigitating reticulum cells probably stimulated with certain immunostimulatory cytokines such as granulocyte/macrophage colony-stimulating factor. It is also suggested that either the second or the third subsets of LnDCs are derived from the first subset. PMID- 9736023 TI - Cerebral amyloid angiopathy: amyloid beta accumulates in putative interstitial fluid drainage pathways in Alzheimer's disease. AB - Cerebral amyloid angiopathy in Alzheimer's disease is characterized by deposition of amyloid beta (Abeta) in cortical and leptomeningeal vessel walls. Although it has been suggested that Abeta is derived from vascular smooth muscle, deposition of Abeta is not seen in larger cerebral vessel walls nor in extracranial vessels. In the present study, we examine evidence for the hypothesis that Abeta is deposited in periarterial interstitial fluid drainage pathways of the brain in Alzheimer's disease and that this contributes significantly to cerebral amyloid angiopathy. There is firm evidence in animals for drainage of interstitial fluid from the brain to cervical lymph nodes along periarterial spaces; similar periarterial channels exist in humans. Biochemical study of 6 brains without Alzheimer's disease revealed a pool of soluble Abeta in the cortex. Histology and immunocytochemistry of 17 brains with Alzheimer's disease showed that Abeta accumulates five times more frequently around arteries than around veins, with selective involvement of smaller arteries. Initial deposits of Abeta occur at the periphery of arteries at the site of the putative interstitial fluid drainage pathways. These observations support the hypothesis that Abeta is deposited in periarterial interstitial fluid drainage pathways of the brain and contributes significantly to cerebral amyloid angiopathy in Alzheimer's disease. PMID- 9736026 TI - Basic fibroblast growth factor is neither necessary nor sufficient for the development of retinal neovascularization. AB - Basic fibroblast growth factor (FGF2) is constitutively expressed in the retina and its expression is increased by a number of insults, but its role in the retina is still uncertain. This study was designed to test the hypothesis that altered expression of FGF2 in the retina affects the development of retinal neovascularization. Mice with targeted disruption of the Fgf2 gene had no detectable expression of FGF2 in the retina by Western blot, but retinal vessels were not different in appearance or total area from wild-type mice. When FGF2 deficient mice were compared with wild-type mice in a murine model of oxygen induced ischemic retinopathy, they developed the same amount of retinal neovascularization. Transgenic mice with a rhodopsin promoter/Fgf2 gene fusion expressed high levels of FGF2 in retinal photoreceptors but developed no retinal neovascularization or other abnormalities of retinal vessels; in the ischemic retinopathy model, they showed no significant difference in the amount of retinal neovascularization compared with wild-type mice. These data indicate that FGF2 expression is not necessary nor sufficient for the development of retinal neovascularization. This suggests that agents that specifically antagonize FGF2 are not likely to be useful adjuncts in the treatment of retinal neovascularization and therapies designed to increase FGF2 expression are not likely to be complicated by retinal neovascularization. PMID- 9736027 TI - Late-onset chronic inflammatory encephalopathy in immune-competent and severe combined immune-deficient (SCID) mice with astrocyte-targeted expression of tumor necrosis factor. AB - To examine the role of tumor necrosis factor (TNF)-alpha in the pathogenesis of degenerative disorders of the central nervous system (CNS), transgenic mice were developed in which expression of murine TNF-alpha was targeted to astrocytes using a glial fibrillary acidic protein (GFAP)-TNF-alpha fusion gene. In two independent GFAP-TNFalpha transgenic lines (termed GT-8 or GT-2) adult (>4 months of age) animals developed a progressive ataxia (GT-8) or total paralysis affecting the lower body (GT-2). Symptomatic mice had prominent meningoencephalitis (GT-8) or encephalomyelitis (GT-2) in which large numbers of B cells and CD4+ and CD8+ T cells accumulated at predominantly perivascular sites. The majority of these lymphocytes displayed a memory cell phenotype (CD44high, CD62Llow, CD25-) and expressed an early activation marker (CD69). Parenchymal lesions contained mostly CD45+ high, MHC class II+, and Mac-1+ cells of the macrophage microglial lineage with lower numbers of neutrophils and few CD4+ and CD8+ T cells. Cerebral expression of the cellular adhesion molecules ICAM-1, VCAM-1, and MAdCAM as well as a number of alpha- and beta-chemokines was induced or upregulated and preceded the development of inflammation, suggesting an important signaling role for these molecules in the CNS leukocyte migration. Degenerative changes in the CNS of the GFAP-TNFalpha mice paralleled the development of the inflammatory lesions and included primary and secondary demyelination and neurodegeneration. Disease exacerbation with more extensive inflammatory lesions that contained activated cells of the macrophage/microglial lineage occurred in GFAP-TNFalpha mice with severe combined immune deficiency. Thus, persistent astrocyte expression of murine TNF-alpha in the CNS induces a late-onset chronic inflammatory encephalopathy in which macrophage/microglial cells but not lymphocytes play a central role in mediating injury. PMID- 9736028 TI - A new role for neurotrophin-3: involvement in the regulation of hair follicle regression (catagen). AB - Nervous system and hair follicle epithelium share a common ectodermal origin, and some neurotrophins (NTs) can modulate keratinocyte proliferation and apoptosis. Therefore, it is reasonable to ask whether NTs are also involved in hair growth control. Here, we show that the expression of NT-3 and its high-affinity receptor, tyrosine kinase C, in the skin of C57BL/6 mice is strikingly hair cycle dependent, with maximal transcript and protein expression seen during spontaneous hair follicle regression (catagen). During catagen, NT-3 and tyrosine kinase C are co-expressed by terminal deoxynucleotidyl transferase-mediated in situ nick end labeling-positive keratinocytes in the club hair and secondary germ. NT-3 overexpressing transgenic mice show precocious catagen development during the postnatal initiation of hair follicle cycling, whereas heterozygous NT-3 knockout (+/-) mice display a significant catagen retardation. Finally, NT-3 stimulates catagen development in organ culture of normal C57BL/6 mouse skin. These observations suggest that the hair follicle is both a source and target of NT-3 and that NT-3/tyrosine kinase C signaling is functionally important in the control of hair follicle regression. Therefore, tyrosine kinase C agonists and antagonists deserve systematic exploration for the management of hair growth disorders that are related to premature (alopecia/effluvium) or retarded catagen (hirsutism/hypertrichosis). PMID- 9736029 TI - Oligodendrocyte apoptosis and primary demyelination induced by local TNF/p55TNF receptor signaling in the central nervous system of transgenic mice: models for multiple sclerosis with primary oligodendrogliopathy. AB - The scientific dogma that multiple sclerosis (MS) is a disease caused by a single pathogenic mechanism has been challenged recently by the heterogeneity observed in MS lesions and the realization that not all patterns of demyelination can be modeled by autoimmune-triggered mechanisms. To evaluate the contribution of local tumor necrosis factor (TNF) ligand/receptor signaling pathways to MS immunopathogenesis we have analyzed disease pathology in central nervous system expressing TNF transgenic mice, with or without p55 or p75TNF receptors, using combined in situ terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling and cell identification techniques. We demonstrate that local production of TNF by central nervous system glia potently and selectively induces oligodendrocyte apoptosis and myelin vacuolation in the context of an intact blood-brain barrier and absence of immune cell infiltration into the central nervous system parenchyma. Interestingly, primary demyelination then develops in a classical manner in the presence of large numbers of recruited phagocytic macrophages, possibly the result of concomitant pro-inflammatory effects of TNF in the central nervous system, and lesions progress into acute or chronic MS-type plaques with axonal damage, focal blood-brain barrier disruption, and considerable oligodendrocyte loss. Both the cytotoxic and inflammatory effects of TNF were abrogated in mice genetically deficient for the p55TNF receptor demonstrating a dominant role for p55TNF receptor-signaling pathways in TNF mediated pathology. These results demonstrate that aberrant local TNF/p55TNF receptor signaling in the central nervous system can have a potentially major role in the aetiopathogenesis of MS demyelination, particularly in MS subtypes in which oligodendrocyte death is a primary pathological feature, and provide new models for studying the basic mechanisms underlying oligodendrocyte and myelin loss. PMID- 9736030 TI - Expression of human herpesvirus-6 antigens in benign and malignant lymphoproliferative diseases. AB - Immunohistochemistry was used to look for the expression of human herpesvirus-6 (HHV-6) antigens in a well characterized series of benign, atypical, and malignant lymphoid lesions, which tested positive for the presence of HHV-6 DNA. A panel of specific antibodies against HHV-6 antigens, characteristic either of the early (p41) or late (p101K, gp106, and gp116) phases of the viral cycle, was applied to the lymphoid tissues from 15 non-Hodgkin's lymphomas, 14 Hodgkin's disease cases, 5 angioimmunoblastic lymphadenopathies with dysproteinemia, 14 reactive lymphadenopathies, and 2 cases of sinus histiocytosis with massive lymphadenopathy (Rosai-Dorfman disease). In lymphomatous tissues, the expression of late antigens was documented only in reactive cells, and mainly in plasma cells. Of interest, the expression of the early p41 antigen was detected in the so-called "mummified" Reed-Sternberg cells, in two Hodgkin's disease cases. In reactive lymphadenopathies, the HHV-6 late antigen-expressing cells were plasma cells, histiocytes, and rare granulocytes distributed in interfollicular areas. In both cases of Rosai-Dorfman disease, the p101K showed an intense staining in follicular dendritic cells of germinal centers, whereas the gp106 exhibited an intense cytoplasmic reaction in the abnormal histiocytes, which represent the histological hallmark of the disease. The expression of HHV-6 antigens is tightly controlled in lymphoid tissues. The lack of HHV-6 antigen expression in neoplastic cells and the limited expression in degenerating Reed-Sternberg cells argue against a major pathogenetic role of the virus in human lymphomagenesis. The detection of a rather unique pattern of viral late antigen expression in Rosai-Dorfman disease suggests a possible pathogenetic involvement of HHV-6 in some cases of this rare lymphoproliferative disorder. PMID- 9736031 TI - Transfer of tumor necrosis factor-alpha to rat lung induces severe pulmonary inflammation and patchy interstitial fibrogenesis with induction of transforming growth factor-beta1 and myofibroblasts. AB - Tumor necrosis factor-alpha is up-regulated in a variety of different human immune-inflammatory and fibrotic pulmonary pathologies. However, its precise role in these pathologies and, in particular, the mechanism(s) by which it may induce fibrogenesis are not yet elucidated. Using a replication-deficient adenovirus to transfer the cDNA of tumor necrosis factor-alpha to rat lung, we have been able to study the effect of transient but prolonged (7 to 10 days) overexpression of tumor necrosis factor-alpha in normal adult pulmonary tissue. We have demonstrated that local overexpression resulted in severe pulmonary inflammation with significant increases in neutrophils, macrophages, and lymphocytes and, to a lesser extent, eosinophils, with a peak at day 7. By day 14, the inflammatory cell accumulation had declined, and fibrogenesis became evident, with fibroblast accumulation and deposition of extracellular matrix proteins. Fibrotic changes were patchy but persisted to beyond day 64. To elucidate the mechanism underlying this fibrogenesis, we examined bronchoalveolar fluids for the presence of the fibrogenic cytokine transforming growth factor-beta1 and tissues for induction of alpha-smooth muscle actin-rich myofibroblasts. Transforming growth factor-beta1 was transiently elevated from day 7 (peak at day 14) immediately preceding the onset of fibrogenesis. Furthermore, there was a striking accumulation of myofibroblasts from day 7, with the most extensive and intense immunostaining at day 14, ie, coincident with the up-regulation of transforming growth factor-beta1 and onset of fibrogenesis. Thus, we have provided a model of tumor necrosis factor-alpha-mediated pulmonary inflammation and fibrosis in normal adult lung, and we suggest that the fibrogenesis may be mediated by the secondary up regulation of transforming growth factor-beta1 and induction of pulmonary myofibroblasts. PMID- 9736033 TI - Complement-mediated killing of microtumors in vitro. AB - Complement-mediated lysis of cancer cells growing in three-dimensional aggregates involves factors that are not associated with the killing of cells in suspension. We have used multicellular tumor spheroids established from breast carcinoma (T47D) and ovarian teratocarcinoma (PA-1) cell lines as models to study complement-mediated destruction of micrometastases and small solid tumors. We found that significant killing of microtumors treated with an antitumor antibody and a specific monoclonal antibody (YTH53.1) against the complement lysis inhibitor protectin (CD59) started to occur after a 1 to 2-hour lag phase. After an overnight incubation, the microtumors became totally infiltrated by the YTH53.1 monoclonal antibody and C1q, whereas C3 and C5b-9 penetrated as a frontier to the peripheral cell layers. A 51Cr release assay showed that during a 24-hour pulsed treatment with complement, 33% of cells in the spheroids were killed, and the average tumor volume decreased by 28%. According to propidium iodide staining, complement exposure resulted in killing and peeling off of the outermost tumor cells. PMID- 9736032 TI - Gelatinases A and B are up-regulated in rat lungs by subacute hyperoxia: pathogenetic implications. AB - Subacute hyperoxia may cause basement membrane disruption and subsequent fibrosis. To test the role of extracellular matrix degradation in hyperoxic damage, we analyzed the expression of gelatinases A and B and tissue inhibitors of metalloproteinases (TIMP)-1 and TIMP-2 in rats exposed to 85% O2. Oxygen exposed rats were studied at 1, 3, 5, and 7 days, and compared with air-breathing rats. Lung mRNAs assayed by Northern and in situ hybridization showed an up regulation of lung gelatinases A and B from the 3rd day on. Gelatinase A was localized in alveolar macrophages and in interstitial and alveolar epithelial cells. Gelatinase B mRNA and protein were localized in macrophages and bronchiolar and alveolar epithelial cells. Increased gelatinase A and B activities were demonstrated in bronchoalveolar lavage. TIMP-1 and TIMP-2 were constitutively expressed, and only TIMP-1 displayed a moderate increase with hyperoxia. To elucidate transcriptional mechanisms for increased gelatinase B expression after hyperoxia, nuclear transcription factor-kappabeta activation was explored. Oxidative stress significantly increased the lung expression of nuclear transcription factor-kappabeta (p65) protein, and nuclear transcription factor kappabeta activation and increased levels of gelatinases A and B were found in isolated type II alveolar cells obtained from hyperoxic rats. Conceivably, subacute hyperoxia induces excessive gelatinase activity, which may contribute to lung damage. PMID- 9736034 TI - Telomerase activity and expression of telomerase RNA component and telomerase catalytic subunit gene in cervical cancer. AB - Telomerase, a ribonucleoprotein complex that includes the telomerase RNA component (hTR) and the telomerase catalytic subunit gene (hTERT) product, has been shown to be activated in the majority of cancer tissues and immortalized cells. To study telomerase activation during the progression of cervical cancer, the expression of hTR and hTERT RNAs in tissues of various stages of cervical cancer was analyzed using the in situ hybridization method and compared with proliferative activity as estimated by Ki-67 immunostaining. To test whether expression of these components is reflected in enzyme activity, we determined the levels of the RNAs in cervical cancer and normal tissues and in primary and immortal keratinocytes by reverse transcription-polymerase chain reaction and RNase protection assays and compared the results to telomerase activities as detected by telomeric repeat amplification protocol assay. In situ hybridization signals of hTR and hTERT were present not only in carcinoma tissues but also in normal epidermal layers. In many adenocarcinoma and fewer squamous cell carcinoma tissues, both signals were focally increased where high proliferative activity was present at the stages of dysplasia/metaplasia, in situ carcinoma, and invasive carcinoma. The level of bTERT, as quantitated by RNase protection assay, was not different between cancer and control tissues or immortal and a subset of primary keratinocytes and did not correlate with telomerase activity. These results indicate that expression of hTR and bTERT is up-regulated in at least a subset of neoplastic cells at an early stage of carcinogenesis and that unidentified factors, such as the modulation or coordination of its protein level with other products, may contribute to the activation of telomerase in cervical cancer. PMID- 9736035 TI - Multistep nature of metastatic inefficiency: dormancy of solitary cells after successful extravasation and limited survival of early micrometastases. AB - In cancer metastasis, only a small percentage of cells released from a primary tumor successfully form distant lesions, but it is uncertain at which steps in the process cells are lost. Our goal was to determine what proportions of B16F1 melanoma cells injected intraportally to target mouse liver 1) survive and extravasate, 2) form micrometastases (4 to 16 cells) by day 3, 3) develop into macroscopic tumors by day 13, and 4) remain as solitary dormant cells. Using in vivo videomicroscopy, a novel cell accounting assay, and immunohistochemical markers for proliferation (Ki-67) and apoptosis (TUNEL), we found that 1) 80% of injected cells survived in the liver microcirculation and extravasated by day 3, 2) only a small subset of extravasated cells began to grow, with 1 in 40 forming micrometastases by day 3, 3) only a small subset of micrometastases continued to grow, with 1 in 100 progressing to form macroscopic tumors by day 13 (in fact, most micrometastases disappeared), and 4) 36% of injected cells remained by day 13 as solitary cancer cells, most of which were dormant (proliferation, 2%; apoptosis, 3%; in contrast to cells within macroscopic tumors: proliferation, 91%; apoptosis/necrosis, 6%). Thus, in this model, metastatic inefficiency is principally determined by two distinct aspects of cell growth after extravasation: failure of solitary cells to initiate growth and failure of early micrometastases to continue growth into macroscopic tumors. PMID- 9736036 TI - ALK expression defines a distinct group of T/null lymphomas ("ALK lymphomas") with a wide morphological spectrum. AB - The t(2;5)(p23;q35) translocation associated with CD30-positive anaplastic large cell lymphoma results in the production of a NPM-ALK chimeric protein, consisting of the N-terminal portion of the NPM protein joined to the entire cytoplasmic domain of the neural receptor tyrosine kinase ALK. The ALK gene products were identified in paraffm sections by using a new anti-ALK (cytoplasmic portion) monoclonal antibody (ALKc) that tends to react more strongly than a previously described ALK1 antibody with the nuclei of ALK-expressing tumor cells after microwave heating in 1 mmol/L ethylenediaminetetraacetic acid buffer, pH 8.0. The ALKc monoclonal antibody reacted selectively with 60% of anaplastic large cell lymphoma cases (60 of 100), which occurred mainly in the first three decades of life and consistently displayed a T/null phenotype. This group of ALK-positive tumors showed a wide morphological spectrum including cases with features of anaplastic large cell lymphoma "common" type (75%), "lymphohistiocytic" (10%), "small cell" (8.3%), "giant cell" (3.3%), and "Hodgkin's like" (3.3%). CD30 positive large anaplastic cells expressing the ALK protein both in the cytoplasm and nucleus represented the dominant tumor population in the common, Hodgkin's like and giant cell types, but they were present at a smaller percentage (often with a perivascular distribution) also in cases with lymphohistiocytic and small cell features. In this study, the ALKc antibody also allowed us to identify small neoplastic cells (usually CD30 negative) with nucleus-restricted ALK positivity that were, by definition, more evident in the small cell variant but were also found in cases with lymphohistiocytic, common, and "Hodgkin's-like" features. These findings, which have not been previously emphasized, strongly suggest that the neoplastic lesion (the NPM-ALK gene) must be present both in the large anaplastic and small tumor cells, and that ALK-positive lymphomas lie on a spectrum, their position being defined by the ratio of small to large neoplastic cells. Notably, about 15% of all ALK-positive lymphomas (usually of the common or giant cell variant) showed a cytoplasm-restricted ALK positivity, which suggests that the ALK gene may have fused with a partner(s) other than NPM. From a diagnostic point of view, detection of the ALK protein was useful in distinguishing anaplastic large cell lymphoma cases of lymphohistiocytic and small cell variants from reactive conditions and other peripheral T-cell lymphoma subtypes, as well as for detecting a small number of tumor cells in lymphohemopoietic tissues. In conclusion, ALK positivity appears to define a clinicopathological entity with a T/null phenotype ("ALK lymphomas"), but one that shows a wider spectrum of morphological patterns than has been appreciated in the past. PMID- 9736037 TI - Loss of p16/INK4A protein expression in non-Hodgkin's lymphomas is a frequent finding associated with tumor progression. AB - The CDKN2A gene located on chromosome region 9p21 encodes the cyclin-dependent kinase-4 inhibitor p16/INK4A, a negative cell cycle regulator. We analyzed p16/INK4A expression in different types of non-Hodgkin's lymphoma to determine whether the absence of this protein is involved in lymphomagenesis, while also trying to characterize the genetic events underlying this p16/INK4A loss. To this end, we investigated the levels of p16/INK4A protein using immunohistochemical techniques in 153 cases of non-Hodgkin's lymphoma, using as reference the levels found in reactive lymphoid tissue. The existence of gene mutation, CpG island methylation, and allelic loss were investigated in a subset of 26 cases, using single-strand conformational polymorphism and direct sequencing, Southern Blot, polymerase chain reaction, and microsatellite analysis, respectively. Loss of p16/INK4A expression was detected in 41 of the 112 non-Hodgkin's lymphomas studied (37%), all of which corresponded to high-grade tumors. This loss of p16/INK4A was found more frequently in cases showing tumor progression from mucosa-associated lymphoid tissue low-grade lymphomas (31 of 37) or follicular lymphomas (4 of 4) into diffuse large B-cell lymphomas. Analysis of the status of the p16/INK4A gene showed different genetic alterations (methylation of the 5' CpG island of the p16/INK4A gene, 6 of 23 cases; allelic loss at 9p21, 3 of 16 cases; and nonsense mutation, 1 of 26 cases). In all cases, these events were associated with loss of the p16/INK4A protein. No case that preserved protein expression contained any genetic change. Our results demonstrate that p16/INK4A loss of expression contributes to tumor progression in lymphomas. The most frequent genetic alterations found were 5'-CpG island methylation and allelic loss. PMID- 9736038 TI - Radiation-induced p53 and p21WAF-1/CIP1 expression in the murine intestinal epithelium: apoptosis and cell cycle arrest. AB - p53-dependent expression of p21WAF-1/CIP1 has been studied in murine intestinal epithelium after exposure to ionizing radiation. In un-irradiated small intestine, neither p53 nor p21WAF-1/CIP1 could be detected by immunohistochemistry. After irradiation (8 Gy), there was a time- and dose dependent increase in the expression of both proteins. In the small bowel, the positional expression of p53 and p21WAF-1/CIP1 was similar but not coincident. Both proteins could be observed throughout the crypts with greatest frequency of expression over the first 15 cell positions, which includes the stem cell population (approximately positions 3 to 5) and the proliferating, transit cell population (approximately positions 5 to 15). p53-positive cells were primarily distributed toward the base of the crypt relative to p21WAF-1/CIP1. Subdivision of the p53-positive cell population revealed that the cells with strongest p53 immunoreactivity were positioned farther toward the base of the crypt, and their distribution was approximately coincident with the frequency distribution of apoptotic cells. Cells that were either weakly or moderately immunoreactive for p53 were located toward the middle of the crypt and were approximately coincident with the distribution of p21WAF-1/CIP1. The numbers of both p53- and p21WAF 1/CIP1-positive cells declined steadily with time, and by 6 days after irradiation there were very few immunoreactive cells to observe. Radiation induced increase in p53 and p21WAF-1/CIP1 expression was not detected in mice homozygously null for p53. Expression of p21WAF-1/CIP1 and incorporation of tritiated thymidine were found to be mutually exclusive. In the large bowel, p21WAF-1/CIP1 and p53 expression were observed along the entire length of the colonic crypts after irradiation (8 Gy), and, unlike in the small intestine, this expression was not only maintained but increased over 72 hours. p21WAF-1/CIP1 immunoreactivity was detected in large intestine epithelium up to 6 days after irradiation. The differential expression of p21WAF-1/CIP1, observed between the large and small bowel and within the small intestinal crypts, is discussed. PMID- 9736040 TI - Selective up-regulation of chemokine IL-8 expression in cystic fibrosis bronchial gland cells in vivo and in vitro. AB - Accumulating evidence suggests that the early pulmonary inflammation pathogenesis in cystic fibrosis (CF) may be associated with an abnormal increase in the production of pro-inflammatory cytokines in the CF lung, even in the absence of infectious stimuli. We have postulated that if baseline abnormalities in airway epithelial cell production of cytokines occur in CF, they should be manifested in the CF bronchial submucosal glands, which are known to express high levels of CFTR (cystic fibrosis transmembrane conductance regulator) protein, the gene product mutated in CF disease. Immunohistochemical analyses showed that CF bronchial submucosal glands in patients homozygous for the deltaF508 deletion expressed elevated levels of the endogenous chemokine interleukin (IL)-8 but not the pro-inflammatory cytokines IL-1beta and IL-6, compared with non-CF bronchial glands. Moreover, basal protein and mRNA expression of IL-8 were constitutively up-regulated in cultured deltaF508 homozygous CF human bronchial gland cells, in an unstimulated state, compared with non-CF bronchial gland cells. Furthermore, the exposure of CF and non-CF bronchial gland cells to an elevated extracellular Cl- concentration markedly increased the release of IL-8, which can be corrected in CF gland cells by reducing the extracellular Cl- concentration. We also found that, in contrast to non-CF gland cells, dexamethasone did not inhibit the release of IL-8 by cultured CF gland cells. The selective up-regulation of bronchial submucosal gland IL-8 could represent a primary event that initiates early airway submucosal inflammation in CF patients. These findings are relevant to the pathogenesis of CF and suggest a novel pathophysiological concept for the early and sustained airway inflammation in CF patients. PMID- 9736039 TI - Prostate stem cell compartments: expression of the cell cycle inhibitor p27Kip1 in normal, hyperplastic, and neoplastic cells. AB - The stem cells of rapidly renewing tissues give rise to transiently proliferating cells, which in turn give rise to postmitotic terminally differentiated cells. Although the existence of a transiently proliferating compartment has been proposed for the prostate, little molecular anatomical evidence for its presence has been obtained to date. We used down-regulation of the cyclin-dependent kinase inhibitor p27Kip1 to identify cells capable of entering the proliferative phase of the cell cycle and, therefore, competent to fulfill the role of the transiently proliferating compartment. We examined the expression of p27Kip1 in relation to its role in the development of prostatic carcinoma. Formalin-fixed paraffin-embedded specimens from matched samples of normal-appearing prostate tissue, benign prostatic hyperplasia, high-grade prostatic intraepithelial neoplasia, primary adenocarcinomas, and pelvic lymph node metastases were evaluated by comparative immunohistochemistry against p27Kip1. In normal appearing prostate epithelium, moderate to strong nuclear staining of p27Kip1 was present in greater than 85% of the terminally differentiated secretory cells. The normal basal cell compartment, believed to contain prostatic stem cells, showed distinctive p27Kip1 expression; acini in epithelial benign prostatic hyperplasia tissue contained more p27Kip1-negative basal cells than acini from non-benign prostatic hyperplasia tissue. A third layer of cells was identified that was sandwiched between the basal cells and the luminal cells, and this layer was consistently p27Kip1 negative. This intermediate layer was accentuated in the periurethral region, as well as in prostate tissue that had been subjected to prior combined androgen blockade. We hypothesize that, on appropriate additional mitogenic stimulation, cells in this layer, and other p27Kip1-negative basal cells, are competent for rapid entry into the cell cycle. Consistent with the fact that cancer cells are capable of cell division, all cases of high-grade prostatic intraepithelial neoplasia and invasive carcinoma also showed down regulation of p27Kip1 as compared with the surrounding normal-appearing secretory cells. In pelvic lymph node metastases, p27Kip1 expression was also reduced. In summary, our results suggest that lack of nuclear p27Kip1 protein may delineate a potential transiently proliferating subcompartment within the basal cell compartment of the human prostate. In addition, these studies support the hypothesis that reduced expression of p27Kip1 removes a block to the cell cycle in human prostate epithelial cells and that dysregulation of p27Kip1 protein levels may be a critical early event in the development of prostatic neoplasia. PMID- 9736041 TI - Alteration of protease expression phenotype of mouse peritoneal mast cells by changing the microenvironment as demonstrated by in situ hybridization histochemistry. AB - Mouse mast cell protease (MMCP) mRNA expression was examined by in situ hybridization histochemistry. Peritoneal mast cells (PMCs) of WBB6F1-(+/+) mice expressed MMCP-2, MMCP-4, MMCP-5, and MMCP-6 mRNAs, but did not express MMCP-1 mRNA. When proliferation of PMCs was induced by culturing them in methylcellulose with T cell-derived cytokines, cells in mast cell colonies expressed MMCP-1 mRNA. These mast cells were transferred to a suspension culture to induce further proliferation. The phenotype of the resulting PMC-derived cultured mast cells was similar to that of bone marrow-derived cultured mast cells. When 10(5) PMC derived cultured mast cells or 10(5) bone marrow-derived cultured mast cells were injected into the stomach wall of mast cell-deficient WBB6F1-W/Wv mice, mast cells that appeared in the mucosa and muscularis propria were similar to mast cells in the stomach of intact WBB6F1-(+/+) mice, indicating the injected cells adapted to a new tissue environment. In contrast, when 10(5) PMCs were injected into the stomach wall of WBB6F1-W/Wv mice, the injected PMCs did not adapt to the mucosa. When 20 PMCs were injected, they proliferated and adapted to the mucosal environment. The present results suggest that PMCs adapt to new environments when proliferation occurs before redifferentiation. PMID- 9736042 TI - Expression of trypsin by epithelial cells of various tissues, leukocytes, and neurons in human and mouse. AB - It has long been believed that trypsin is normally synthesized only in the pancreas. In the present study, expression of trypsin in human and mouse nonpancreatic tissues was examined. Northern blot analysis of normal human tissues indicated that the trypsin gene is expressed at high levels in the pancreas and spleen and considerably in the small intestine. However, in situ hybridization and immunohistochemistry demonstrated that trypsin is widely expressed in epithelial cells of the skin, esophagus, stomach, small intestine, lung, kidney, liver, and extrahepatic bile duct, as well as splenic and neuronal cells. In the spleen, trypsin message was detected in macrophages, monocytes, and lymphocytes in the white pulp. In the brain, it was detected in the nerve cells of the hippocampus and cerebral cortex. Analysis by gelatin zymography confirmed the presence of a latent or an active form of trypsin in various normal mouse tissues. Reverse transcription-polymerase chain reaction analysis also confirmed the expression of trypsin genes in the spleen, liver, kidney, and brain of normal mice. Such a broad distribution of trypsin suggests its general roles in the maintenance of normal epithelial cell functions, the immune defense system, and the central nervous system. PMID- 9736043 TI - Differential expression and origin of membrane-type 1 and 2 matrix metalloproteinases (MT-MMPs) in association with MMP2 activation in injured human livers. AB - Matrix metalloproteinase-2 (MMP2) activation is associated with basement membrane remodeling that occurs in injured tissues and during tumor invasion. The newly described membrane-type MMPs (MT-MMPs) form a family of potential MMP2 activators. We investigated the localization and steady-state levels of MT1-MMP and MT2-MMP mRNA, compared with those of MMP2 and tissue inhibitor of MMP-2 in 22 hepatocellular carcinomas, 12 liver metastases from colonic adenocarcinomas, 13 nontumoral samples from livers with metastases, 10 benign tumors, and 6 normal livers. MMP2 activation was analyzed by zymography in the same series. The expression of MT1-MMP mRNA and the activation of MMP-2 were increased in hepatocellular carcinomas, metastases, and cholestatic nontumoral samples. MT2 MMP mRNA was rather stable in the different groups. MT1-MMP mRNA levels, but not MT2-MMP mRNA, correlated with MMP-2 and tissue inhibitor of MMP-2 mRNA levels and with MMP2 activation. In situ hybridization showed that MT1-MMP mRNA was expressed in stromal cells, and MT2-MMP mRNA was principally located in both hepatocytes and biliary epithelial cells. Consistently, freshly isolated hepatocytes expressed only MT2-MMP mRNA, and culture-activated hepatic stellate cells showed high levels of MT1-MMP mRNA. These results indicate that in injured livers, MMP2 activation is related to a coordinated high expression of MMP2, tissue inhibitor of MMP-2, and MT1-MMP. Furthermore, the finding of a preferential expression of MT2-MMP in hepatocytes, together with our previous demonstration that the activation of stellate cell-derived MMP2 in co-culture requires interactions with hepatocytes (Am J Pathol 1997, 150:51-58), suggests that parenchymal cells might play a pivotal role in the MMP2 activation process. PMID- 9736044 TI - Transforming growth factor alpha levels in liver and blood correlate better than hepatocyte growth factor with hepatocyte proliferation during liver regeneration. AB - Transforming growth factor alpha (TGFalpha) and hepatocyte growth factor (HGF) are mitogens for hepatocytes in vitro and in vivo, produced by hepatocytes or nonparenchymal cells such as stellate cells in the liver. It is still uncertain whether TGFalpha and HGF are essential for liver regeneration. To assess the role of these growth factors in liver regeneration, their circulating and hepatic levels were studied in various rat models of liver regeneration. Hepatic and plasma HGF levels were increased with increased number of mitotic hepatocytes in rats after partial hepatectomy or carbon tetrachloride intoxication. However, hepatic HGF levels were decreased despite an increased number of mitotic hepatocytes and increased or unchanged plasma HGF levels in rats given phenobarbital and in rats after dimethylnitrosamine intoxication, which can induce hepatic necrosis after apoptosis of hepatic stellate cells. In contrast, hepatic and serum TGFalpha levels were increased in all of the models. In sham operated rats with no increased number of mitotic hepatocytes, hepatic and circulating levels of HGF were increased, whereas those levels of TGFalpha were unchanged. The results indicate that TGFalpha levels in liver and blood more closely correlate with hepatocyte mitogenesis than HGF levels. PMID- 9736045 TI - Active cyclin A-CDK2 complex, a possible critical factor for cell proliferation in human primary lung carcinomas. AB - Expression of cyclins A and E and cyclin-dependent kinase 2 (CDK2) was examined immunohistochemically in 190 cases of human lung carcinoma. Cyclin A and CDK2 were expressed in the majority of squamous cell carcinomas, small cell carcinomas, and large cell carcinomas, but in significantly fewer cases of adenocarcinomas. Cyclin E was expressed in a minority of all subtypes. In particular, well differentiated cells in squamous cell carcinoma stained positively for cyclin E; in contrast, cyclin A was expressed in the nonkeratinized proliferating areas of the tumor nests. Immunoblotting revealed that all these proteins were expressed at higher levels in tumor tissues than in adjacent normal tissues. Immunoprecipitation also revealed higher levels of cyclin A and cyclin E associated with CDK2 in tumor tissues. Furthermore, tumor tissues which exhibited higher cyclin A and CDK2 expression also had higher CDK2 kinase activity. However, cyclin E-associated kinase activity was barely detectable even in tumor samples exhibiting higher cyclin E expression. Consistent with these data, elevated expression of cyclin A correlated to shorter survival periods in contrast to expression of cyclin E, which correlated to longer survival periods. These results suggest that in human lung carcinomas, elevated expression of active cyclin A-CDK2 complexes with associated higher CDK2 kinase activity is critical for promoting cell cycle progression and unrestrained proliferation of tumor cells and can be a predictive marker for patients' prognosis. On the other hand, immunohistochemical detection of cyclin E-CDK2 reflects accumulation of inactive forms of protein complexes, implying differentiation or senescence of the tumor and the better prognosis. PMID- 9736046 TI - Correlation of reduction in MRP-1/CD9 and KAI1/CD82 expression with recurrences in breast cancer patients. AB - MRP-1/CD9, KAI1/CD82, and ME491/CD63, have been reported to be associated with the metastatic potential of solid tumors. The aim of this study was to determine whether their expression in tumor tissues is a useful indicator for prognosis in breast cancer patients. We studied 109 breast cancer patients who underwent surgery. Quantitative reverse transcription-polymerase chain reaction analysis was performed to evaluate the expression of these genes. The results were confirmed with immunohistochemistry. All of the carcinomas were ME491/CD63 positive. Thirty-six tumors were MRP-1/CD9 negative. The disease-free survival rate and the 5-year survival rate of patients with MRP-1/CD9-negative tumors were both significantly lower than that in patients with MRP-1/ CD9-positive tumors (P = 0.0005 and P = 0.0380, respectively). Sixty-five tumors were KAI1/CD82 negative. The disease-free survival rate of patients with KAI1/CD82-negative tumors was significantly lower than that of patients with KAI1/CD82-positive tumors (P = 0.0065). Cox regression analysis demonstrated that MRP-1/CD9 status (P = 0.0016) and KAI1/CD82 status (P = 0.0234) were useful indicators for the disease-free survival of breast cancer patients. The disease-free survival rate and 5-year survival rate of patients with either MRP-1/CD9-negative or KAI1/CD82 negative tumors were both significantly lower than patients who were positive for both genes (P = 0.0003 and P = 0.0292, respectively). The expression of MRP-1/CD9 and KAI1/CD82 genes are useful indicators of a poor prognosis in breast cancer patients. PMID- 9736047 TI - DNA copy number changes in development and progression in leiomyosarcomas of soft tissues. AB - DNA copy number changes were investigated in 29 leiomyosarcomas by comparative genomic hybridization. The most frequent losses were detected in 10q (20 cases, 69%) and 13q (17 cases, 59%). The most frequent gains were detected in 17p (16 cases, 55%). The most frequent high-level amplifications were detected in 17p (7 cases, 24%) and 8q (6 cases, 21%). A total of 137 losses and 204 gains were detected. Small tumors (less than 5 cm in diameter) displayed fewer changes per sample (3 to 11; mean, 7) than the other tumors (4 to 22; mean, 13). There was an increase in the number of gains from small tumors (mean, 4) to very large tumors (>20 cm; mean, 10). However, the number of losses was similar in small, large, and very large tumors (mean, 4.5). Tumor size-related aberrations were observed. Gains in 16p were detected in all small tumors but were infrequent in large and very large tumors (27% and 11%, respectively). Similarly, gains and high-level amplifications in 17p were more common in small (80%) than in very large tumors (33%). Gains in 1q, 5p, 6q, and 8q were not seen in any of the small tumors but were detected in large and very large tumors. Gains in 6q and 8q occurred in 8 of 9 cases (89%) of very large tumors, 5 of them with a high-level amplification in 8q. PMID- 9736049 TI - Geographically distinct HHV-8 DNA sequences in Saudi Arabian Iatrogenic Kaposi's sarcoma lesions. AB - A new member of the gamma-herpesvirus family, HHV-8 (also known as Kaposi's sarcoma (KS)-associated herpesvirus), has been linked to KS and body cavity-based lymphoma. Other members of this family, eg, Epstein-Barr virus, were originally thought to have only one strain, but subsequent analysis revealed different strains correlating to cellular patterns of infectivity and geographical location. To determine whether multiple strains of HHV-8 exist, we compared DNA sequences among KS and body cavity-based lymphoma-derived HHV-8 and examined differences in HHV-8 subgroups between American and Saudi Arabian iatrogenic KS patients. Samples were analyzed by polymerase chain reaction using multiple primer sets to five different open reading frames from HHV-8, and DNA sequencing was performed. HHV-8 DNA was present in all of our KS and body cavity-based lymphoma samples by polymerase chain reaction. HHV-8 DNA was detected in each body cavity-based lymphoma sample using a majority of the primers, whereas only two primer sets consistently amplified HHV-8 DNA derived from KS lesions. DNA sequencing within open reading frames 26 and 27 indicate the existence of at least three variants of HHV-8, with the majority of iatrogenic KS patients in Saudi Arabia containing unique nucleotide changes that may define a distinct, previously unidentified subgroup we term SA, whereas those from America were of Group A or B. Thus, although the sequencing data within open reading frames 26 and 27 did not permit discrimination between patients with lymphoma versus KS disease processes, HHV-8 derived from Saudi Arabian KS lesions were shown to have a distinct nucleotide sequence not seen in any of the other clinical samples examined. PMID- 9736048 TI - Microdissecting the genetic events in nephrogenic rests and Wilms' tumor development. AB - Nephrogenic rests are precursor lesions associated with about 40% of Wilms' tumors. This study identifies genetic steps occurring in the development of Wilms' tumor. Thirty-four Wilms' tumors with nephrogenic rests and/or areas of anaplasia were microdissected from paraffin sections to determine whether and at what stage loss of heterozygosity (LOH) occurred, using polymerase chain reaction based polymorphic markers at 11p13, 11p15, and 16q. LOH at these loci have been identified in Wilms' tumors and are associated with identified or putative tumor suppressor genes. Three cystic nephromas/cystic partially differentiated nephroblastomas were also examined. LOH was detected in six cases at 11p13 and in six cases at 11p15, and two of these cases had LOH at both loci. All intralobar rests showing LOH also showed LOH in the tumor. A case with a small perilobar rest showed LOH of 11p13 only in the tumor. Five cases showing LOH at 16q were identified (this was identified only in the tumor, and not in the associated rest), and three of these had recurrence of the tumor. Two cases had a WT1 mutation (one germline and the other somatic), as well as LOH in both the intralobar rest and the tumor. A cystic partially differentiated nephroblastoma showed loss at 11p13 and 11p15, as well as at 16q. This study suggests that LOH at 11p13 and 11p15 and WT1 mutations are early events but that LOH at 16q occurs late in the pathogenesis of Wilms' tumor. Intralobar and perilobar nephrogenic rests are known to have different biological behaviors, and this study suggests that they are genetically different. A multistep model of Wilms' tumor pathogenesis is supported by these findings. PMID- 9736050 TI - Brain trauma induces massive hippocampal neuron death linked to a surge in beta amyloid levels in mice overexpressing mutant amyloid precursor protein. AB - Although brain trauma is a risk factor for Alzheimer's disease, no experimental model has been generated to explore this relationship. We developed a model of brain trauma in transgenic mice that overexpress mutant human amyloid precursor protein (PDAPP) leading to the appearance of Alzheimer's disease-like beta amyloid (Abeta) plaques beginning at 6 months of age. We induced cortical impact brain injury in the PDAPP animals and their wild-type littermates at 4 months of age, ie, before Abeta plaque formation, and evaluated changes in posttraumatic memory function, histopathology, and regional tissue levels of the Abeta peptides Abeta1-40 and Abeta1-42. We found that noninjured PDAPP mice had impaired memory function compared to noninjured wild-type littermates (P < 0.01) and that brain injured PDAPP mice had more profound memory dysfunction than brain-injured wild type littermates (P < 0.001). Although no augmentation of Abeta plaque formation was observed in brain-injured PDAPP mice, a substantial exacerbation of neuron death was found in the hippocampus (P < 0.001) in association with an acute threefold increase in Abeta1-40 and sevenfold increase in Abeta1-42 levels selectively in the hippocampus (P < 0.01). These data suggest a mechanistic link between brain trauma and Abeta levels and the death of neurons. PMID- 9736051 TI - Cutaneous silent period. PMID- 9736052 TI - The role of axonal ion conductances in diabetic neuropathy: a review. AB - Diabetic neuropathy is a common complication in diabetes mellitus. Diabetic neuropathy is accompanied by alterations in axonal excitability, which can lead to either "positive" (paresthesia, dysesthesia, pain) and/or "negative" (hypesthesia, anesthesia) symptoms. The mechanisms underlying these alterations in axonal excitability are not well understood. Clinical tests reveal reduced nerve conduction velocity and axonal loss, but fail to explain nerve excitability. Many different factors have been suggested in relation to the pathophysiology of diabetic neuropathy. There are probably as many factors as there are different clinical pictures in diabetic neuropathy. Nevertheless, it seems that hyperglycemic hypoxia is mainly responsible for the electrophysiological changes seen in damaged diabetic nerves. This article summarizes experimental data indicating that a dysfunction of ion conductances, especially voltage-gated ion channels, could contribute to abnormalities in the generation and/or conduction of action potentials in diabetic neuropathy. PMID- 9736053 TI - Mechanisms underlying spinal motor neuron excitability during the cutaneous silent period in humans. AB - The transient suppression of muscle contraction during the cutaneous silent period (CSP) could be produced either through postsynaptic inhibition of motoneurons or through presynaptic inhibition of the excitatory inputs to motoneurons that sustain voluntary contraction. We sought to delineate the mechanisms underlying the CSP in hand muscles by measuring changes in H-reflexes and motor-evoked potentials (MEPs) produced by transcranial magnetic stimulation (TMS) during the CSP in 10 healthy volunteers. H-reflexes and MEPs both measure the excitability of the motoneuron pool and activate similar subpopulations of motoneurons through different pathways. Inhibition of H-reflexes and MEPs of similar size was maximal at the midpoint of the CSP and gradually returned to baseline. The similar time course of recovery suggests that the H-reflex and MEP are affected by inhibition at a common site, most likely postsynaptic inhibition of the motoneurons. PMID- 9736054 TI - The force-frequency relationship is altered in regenerating and senescent rat skeletal muscle. AB - Maximal tetanic tension was elicited at 200, 150, and 150 Hz in control tibialis anterior muscles and at 150, 100, and 100 Hz in 14-day regenerating muscles of young (3 months), adult (18 months), and old (31 months) Fischer 344/Brown Norway F1 rats, respectively. In contrast to young rats, increasing stimulation frequency from 50 to 150 Hz did not elicit significantly greater tetanic tension in control or regenerating muscles of old rats. At higher stimulation frequencies, tetanic fade was prevalent in control and regenerating muscles of adult (250-300 Hz) and old rats (200-300 Hz), but was only present at 14 days of recovery in regenerating muscles of young rats (300 Hz). The decreased efficacy of rehabilitative and physical medicine procedures in adult and elderly patients who have suffered skeletal muscle injury could be explained, in part, by the postulate that tetanic fade is indicative of inadequate synaptic transmission. PMID- 9736055 TI - Electrophysiological studies in the Guillain-Barre syndrome: distinguishing subtypes by published criteria. AB - Guillain-Barre syndrome (GBS) is recognized clinically by the presence of acute, rapidly progressive weakness, areflexia, and albuminocytological dissociation in cerebrospinal fluid. Although GBS was initially considered to be primarily an acute inflammatory demyelinating polyneuropathy (AIDP), several other subtypes have been recognized: acute motor axonal neuropathy (AMAN), acute motor-sensory axonal neuropathy (AMSAN), and Fisher syndrome (FS). Because each of these subtypes may have an independent immunopathogenesis and, therefore, may require selective treatments in the future, recognition of these subtypes is important. When using nerve conductions to classify the subtypes, the most easily and confidently identified subtype is AIDP. Therefore, most electrodiagnostic criteria have attempted to identify demyelination in this acute setting, in which physiology is constantly changing. In a single well-defined GBS population, we compared the various published criteria for demyelination in GBS. We reviewed charts of 43 patients with GBS between 1991 and 1996. Applying six available criteria sets, the number of patients categorized as having AIDP ranged from 21% to 72%. Until investigators can agree on a single set of criteria, considerable variability will continue to exist when identifying cases of AIDP. PMID- 9736056 TI - Disproportionate loss of thin filaments in human soleus muscle after 17-day bed rest. AB - Previously we reported that, after 17-day bed rest unloading of 8 humans, soleus slow fibers atrophied and exhibited increased velocity of shortening without fast myosin expression. The present ultrastructural study examined fibers from the same muscle biopsies to determine whether decreased myofilament packing density accounted for the observed speeding. Quantitation was by computer-assisted morphometry of electron micrographs. Filament densities were normalized for sarcomere length, because density depends directly on length. Thick filament density was unchanged by bed rest. Thin filaments/microm2 decreased 16-23%. Glycogen filled the I band sites vacated by filaments. The percentage decrease in thin filaments (Y) correlated significantly (P < 0.05) with the percentage increase in velocity (X), (Y = 0.1X + 20%, R2 = 0.62). An interpretation is that fewer filaments increases thick to thin filament spacing and causes earlier cross bridge detachment and faster cycling. Increased velocity helps maintain power (force x velocity) as atrophy lowers force. Atrophic muscles may be prone to sarcomere reloading damage because force/microm2 was near normal, and force per thin filament increased an estimated 30%. PMID- 9736057 TI - Recruitment stability in masseter motor units during isometric voluntary contractions. AB - Recruitment of single motor units (SMUs) of the masseter muscle was studied using macro representation (MacroRep) as the indicator of motor unit size. When subjects followed a slow isometric force ramp, units were usually recruited in order of MacroRep size. However, pooling the data from repeated ramps in the same subject resulted in a weak relationship between MacroRep size and force recruitment threshold, probably due to marked variations in the relative contributions of the jaw muscles, and varying levels of cocontraction, in the development of total bite force in each ramp. The force recruitment thresholds of individual SMUs showed marked variability, but recruitment threshold stability was improved when expressed as a percentage of maximum surface electromyographic (SEMG) activity in the ipsilateral masseter. Therefore the SEMG recruitment threshold was concluded to be a more stable and accurate indicator of the SMU's position in the recruitment hierarchy in a given muscle. It was concluded that SMUs in masseter are recruited according to the size principle, and that when investigating recruitment in jaw muscles, SEMG recruitment threshold should be used in preference to force recruitment threshold. PMID- 9736058 TI - Ryanodine receptor gene expression thymomas. AB - Ryanodine receptor (RyR) antibodies are present in sera of myasthenics with thymoma, and their titer correlates with morbidity and mortality. We investigated whether skeletal muscle RyR expression in thymic tissues could be the source of immune sensitization to the RyR. Skeletal muscle RyR gene expression was investigated using reverse transcription followed by semiquantitative polymerase chain reaction. Hyperplastic and normal thymuses expressed significant levels of RyR, but RyR gene transcripts was statistically less likely in thymomas than in hyperplastic and normal thymus (P < 0.05). The presence of RyR transcripts in thymomas did not correlate with myasthenic manifestations, thymic pathology, or serum RyR antibodies. We conclude that the skeletal muscle RyR in thymoma is not the inciting antigen for immune sensitization to RyR epitopes in thymoma associated myasthenia gravis. PMID- 9736059 TI - Cortical excitability in patients with essential tremor. AB - We used transcranial magnetic stimulation in 10 patients with essential tremor and 8 matched healthy subjects. A round stimulating coil was placed over the vertex and electromyographic activity was recorded from the first dorsal interosseous muscle. Paired transcranial stimuli were delivered at interstimulus intervals of 3, 5, 20, 100, 150, and 200 ms. The intensity of the conditioning stimulus was 80% of motor threshold at short and 150% at long interstimulus intervals (ISIs). We also measured the silent period obtained after a single magnetic pulse delivered at 150% of motor threshold during a submaximal muscle contraction. Patients and controls had similar motor threshold and similar latencies. Paired magnetic stimuli given at short and long ISIs at rest, and during a voluntary muscle contraction, elicited similar responses in both groups. The silent period evoked by transcranial magnetic stimulation had a similar duration in patients with ET and controls. In conclusion, these findings suggest that patients with essential tremor have normal cortical motor area excitability. PMID- 9736060 TI - Task-dependent facilitation of motor evoked potentials during dynamic and steady muscle contractions. AB - Task-dependent differences in the facilitation of motor evoked potentials (MEPs) following cortex stimulation were studied in a proximal (deltoid) and a distal muscle (abductor digiti minimi; ADM) in 23 healthy subjects during both dynamic and steady contractions of the target muscle under isometric and under nonisometric conditions. In the deltoid, MEP amplitudes were significantly greater if stimulation was performed during dynamic contractions than during steady contractions, despite equal background electromyographic levels just prior to the stimulus. The same task-specific extra facilitation of deltoid MEP amplitudes was also found with magnetic stimulation of the brain stem instead of the cortex in 3 subjects. In the ADM, no such task-dependent extra facilitation of MEPs during dynamic contractions was found. It is concluded that in the deltoid, during dynamic contractions, a greater proportion of the spinal motoneurons is close to depolarization threshold (greater "subliminal fringe") whereas the number of firing motoneurons is similar to that during steady contraction. The lack of task-dependent extra facilitation of MEPs in the ADM is explained by the predominant recruitment principle for force gradation in small hand muscles, which is in contrast to the predominant frequency principle used in proximal muscles. PMID- 9736061 TI - Mosaic expression of two dystrophins in a boy with progressive muscular dystrophy. AB - A boy with a Becker muscular dystrophy (BMD) phenotype presented unique muscular dystrophin expression. Western blot analysis showed the presence of two dystrophins of different sizes, i.e., a 400-kDa dystrophin and a 500-kDa form. An immunofluorescent study revealed mosaic expression of these dystrophins in the sarcolemma, with matching alpha-sarcoglycan and beta-dystroglycan staining patterns. DNA and RNA analysis did not reveal any mutation in the dystrophin gene, and the karyotype was normal. PMID- 9736062 TI - Neurophysiologic evaluation of lower motor neuron damage in tetraplegia. AB - We quantitatively investigated the extent of damage to motor neurons in tetraplegic subjects. Numbers of motor units in the patients were significantly lower for thenar, wrist extensor, and biceps brachii as compared to controls. Reduction in counts occurred even when M-response amplitudes were normal. Standard electromyography suggested a surprising frequency of lower motor neuron dysfunction below the level of injury. These results confirm previous reports and add data on motor units in the biceps brachii. PMID- 9736063 TI - Effect of stimulator orientation on F-wave persistence. AB - Most electrodiagnosis texts advocate cathode distal stimulation (CDS) for nerve conduction, but suggest cathode proximal stimulation (CPS) for F waves, because of anodal block. We postulated that CDS and CPS elicit F waves of similar persistence. We studied 657 (207 median, 204 ulnar, 136 tibial, and 110 peroneal) nerves in 225 consecutive subjects. In the median nerve, CDS elicited F waves of slightly greater persistence than CPS. Stimulator orientation did not affect F wave persistence in the remaining nerves. PMID- 9736064 TI - Relapses in the Guillain-Barre syndrome after treatment with intravenous immune globulin or plasma exchange. AB - To clarify the question of whether Guillain-Barre syndrome (GBS) patients treated with intravenous immune globulin (i.v.IG) relapse at a higher frequency than those treated with plasma exchange (PE), 54 patients with GBS were studied retrospectively. A higher frequency of relapses was noted in the PE-treated patients than in those receiving i.v.IG. The presence of an associated medical condition correlated with an increased risk of relapses, while earlier onset of treatment resulted in a decrease of relapses of GBS. This study found no support for prior suggestions of increased relapses in patients with GBS treated with i.v.IG as opposed to those treated with PE. PMID- 9736065 TI - Compound muscle action potential cartography of an accessory peroneal nerve. AB - In daily practice, accessory peroneal nerves (APNs) are detected in less than the 18-25% of legs, as revealed by systematic searches. In one APN case, compound muscle action potential cartography showed that the APN was only apparent when the recording electrode was placed over a small lateral region of the extensor digitorum brevis muscle. Effects of recording site can explain why many APNs go unrecognized. PMID- 9736066 TI - Transient weakness and compound muscle action potential decrement in myotonia congenita. AB - Twenty-five Turkish patients with recessive myotonia congenita (RMC), 16 of whom had genetic confirmation, were studied. Nineteen had transient weakness. In the upper extremities, onset age of transient weakness was usually in the early teens. All untreated RMC patients had a compound muscle action potential decrement of > or =25%, usually above 50%, with repetitive nerve stimulation at 10/s for 5 s. Patients with other nondystrophic diseases with myotonia, except 1 patient with dominant myotonia congenita, had no transient weakness and a CMAP decrement below 25%. PMID- 9736068 TI - Axonal and demyelinating neuropathy with reversible proximal conduction block, an unusual feature of vitamin B12 deficiency. AB - We report a 35-year-old patient with megaloblastic anemia who presented with features of subacute combined degeneration of the cord. Electrophysiological studies showed features of axonal neuropathy. In addition, there was evidence of prominent focal proximal conduction block in several nerves. After treatment with cyanocobalamin the neuropathy improved, and the peripheral nerve conduction block detected earlier disappeared. Reversible nerve conduction block as a feature of vitamin B12 deficiency in man, to our knowledge, has not been reported in literature, so far. PMID- 9736067 TI - Firing rate analysis using decompostion-enhanced spike triggered averaging in the quadriceps femoris. AB - Electromyographic signals detected from the quadriceps femoris during various constant force contractions were decomposed to identify individual motor unit discharges and mean firing rates (FRs). Subject and group mean FRs were calculated for each force level. Mean FR values and FR variability increased with force. Individual, subject, and group mean FRs showed slight increases until 30% of maximum voluntary contraction and larger increases thereafter. Findings are discussed in relation to motor unit recruitment, frequency modulation, and fatigue. PMID- 9736069 TI - Postnatal development of nitric oxide synthase activity in fast and slow muscles of the rat. AB - Nitric oxide synthase (NOS) activity was measured in extensor digitorum longus (EDL) and soleus muscles during postnatal development in the rat. At 1 and 2 weeks of age, similar low levels were found in both muscles. After 2 weeks, activity increased significantly only in EDL. Adult NOS activity was significantly higher in EDL than soleus. Thus, the preferential expression of NOS in fast muscle only occurs once the adult pattern of motor activity is established. PMID- 9736070 TI - Fibrillations in lumbosacral paraspinal muscles of normal subjects. AB - Although paraspinal muscle fibrillations and positive sharp waves (PSWs) are used to help identify neuromuscular disorders, the frequency of these abnormalities in normal subjects is uncertain. We performed lumbosacral paraspinal electromyography in 65 normal subjects. Twenty-seven (42%) had fibrillations and/or PSWs, with the prevalence of these findings increasing with age (r = 0.830, P = 0.040). These data suggest isolated fibrillations and PSWs in lumbosacral paraspinal muscles, especially of older subjects, are nonspecific findings. PMID- 9736071 TI - Focal atrophy of the multifidus muscle in lumbosacral radiculopathy. AB - A patient with compelling clinical and electrodiagnostic evidence of a right L5 radiculopathy had focal atrophy of the multifidus at the appropriate level, which served to confirm the radicular nature of the process. The multifidus muscles are innervated by a single root, in contrast to the polysegmental innervation of the rest of the paraspinal muscle mass. Imaging studies may complement needle electromyography in the evaluation of this important structure. PMID- 9736072 TI - Collision technique in Martin-Gruber anastomosis. PMID- 9736073 TI - A case of eosinophilic polymyositis complicated by myasthenia gravis. PMID- 9736074 TI - Small-cell carcinoma of the lung presenting with paraneoplastic peripheral nerve microvasculitis and optic neuropathy. PMID- 9736075 TI - Successful treatment of orbital myositis with intravenous immunoglobulins. PMID- 9736076 TI - Implications of the guidelines for the management of severe head injury for the practicing neurosurgeon. PMID- 9736077 TI - Early surgery and other indicators influencing the outcome of war missile skull base injuries. AB - BACKGROUND: The aim of this study was to analyze the effect of early surgical management protocol and other important clinical features on the prognosis of patients suffering from war missile skull base injuries. METHODS: Twenty-one patients who suffered from war missile skull base injuries were analyzed in this study. The wounds were mainly caused by shells and/or bullets. Craniotomy represented the standard treatment in all patients. Investigated clinical features included Glasgow Coma Scale score on admission, the mode and the extent of brain injury, time to patient admission to hospital, and the presence of an intracranially retained foreign body. The prognostic importance of complications such as infection, intracranial hemorrhage, cerebrospinal fluid leak, and epileptic seizures was also investigated. RESULTS: The outcome of 21 skull base injuries was as follows: death in seven patients, vegetative state in three, severe disability in two, moderate disability in seven, and good recovery in two patients. The clinical characteristics that implied favorable outcome were: Glasgow Coma Scale score greater than 12, location of injury in the anterior cranial fossa, time to admission shorter than 1 hour, and absence of an intracranially retained foreign body and postoperative complications. The statistical significance of those predictors was at the level of p < 0.001 in all cases. CONCLUSIONS: Although the wounds were associated with a high mortality rate, this study showed that there are major differences in prognosis of patients with war missile skull base injuries with respect to certain presenting clinical features. PMID- 9736078 TI - Chronic pain syndrome. PMID- 9736079 TI - Functional neuromuscular stimulator for short-distance ambulation by certain thoracic-level spinal-cord-injured paraplegics. AB - BACKGROUND: Functional Neuromuscular Stimulation (FNS) for unbraced short distance ambulation by traumatic complete/near-complete T4 to T12 paraplegics is based on work by Graupe et al (1982), Kralj et al (1980), Liberson et al (1961), and others. This paper discusses methodology, performance, training, admissibility criteria, and medical observations for FNS-ambulation using the Parastep-I system, which is the first and only such system to have received FDA approval (1994) and which emanated from these previous works. METHOD: The Parastep system is a transcutaneous non-invasive and microcomputerized electrical stimulation system built into a Walkman-size unit powered by eight AA batteries that is controlled by finger-touch buttons located on a walker's handbars for manual selection of stimulation menus. The microcomputer shapes, controls, and distributes trains of stimulation signals that trigger action potentials in selected peripheral nerves. Walker support is used for balance. The patient can don the system in under 10 minutes. At least 32 training sessions are required. RESULTS: Approximately 400 patients have used the Parastep system, essentially all achieving standing and at least 30 feet of ambulation, with a few reaching as much as 1 mile at a time. Recent literature presents data on the medical benefits of using the Parastep system-beyond the exercise benefits of short distance ambulation at will-such as increased blood flow to the lower extremities, lower HR at subpeak work intensities, increased peak work capability, reduced spasticity, and psychological benefits. CONCLUSIONS: We believe that the Parastep FNS system, which is presently commercially available by prescription, is easily usable for independent short-distance ambulation. We believe that its exercise benefits and its other medical and psychological benefits, as discussed, make it an important option for thoracic-level traumatic paraplegics. PMID- 9736080 TI - Topical irrigation with polymyxin and bacitracin for spinal surgery. AB - BACKGROUND: The purpose of the present study was to evaluate constant irrigation with saline containing 50,000 units each of polymyxin and bacitracin in a regimen of antimicrobial prophylaxis for clean spinal surgery at two community hospitals with a zero infection rate. METHODS: The focus was on the bactericidal effects of prophylactic topical antibiotics by assessing random contamination in neurosurgical wounds from: 1) the flora of the integument and nares of the operating team, 2) the surgical apparel, 3) the patient's skin, 4) air-borne organisms in the operating theater, and 5) the surgeon's gloves. RESULTS: Based on individual biotyping of bacteria and antimicrobial sensitivity testing, no consistent source or pattern could be uncovered for the organisms recovered from the operative site. Relying on longitudinal data, the incidence of intraoperative bacterial growth with continuous saline lavage was reduced from 64 to 4% when the combination of topical polymyxin and bacitracin was added. CONCLUSIONS: Although the virtual elimination of bacterial growth in the surgical site was accomplished, the efficacy of topical antibiotics in the prevention of wound infection remains unproven. PMID- 9736081 TI - Microsurgical anatomy of the dorsal cervical nerve roots and the cervical dorsal root ganglion/ventral root complexes. AB - BACKGROUND: It is known that a "dissociated motor loss" of the deltoid muscle can occur with disconcerting frequency after cervical spine surgery. The etiology of this entity is in question. We conducted an anatomic study to identify anatomic factors that might predispose C5 to injury. METHODS: We studied 128 dorsal cervical nerves and root ganglion/ventral root complexes in 10 adult cadavers. At each cervical level the following data were recorded: number of rootlets, range of width of rootlets, length of DREZ, cranial angles of the superior and inferior rootlets with the spinal cord, length of the superior and inferior rootlets, dimensions of the foramina, dimensions of the dorsal root, dimensions of the dorsal root ganglion (DRG)/ventral root (VR) complex, and the blood supply to the DRG. The histology at the site of compression was also examined. Statistical analysis was conducted using the single factor-repeated measures analysis of variance. RESULTS: We found that, 1) the C5 superior dorsal rootlets angle less inferiorly from the cervical cord than the other dorsal cervical roots (p=0.001), 2) the majority of the DRG/VR complexes from C3 to C6 were compressed by the vertebral artery (73%), 3) the C5 DRG/VR complex was compressed to the greatest extent (77.6%, p=0.3519), and 4) the ganglionic artery was more frequent at C4, C5, and C6. CONCLUSION: To our knowledge, the second finding has not been reported previously. The first and third findings may help explain why C5 is more vulnerable to injury. PMID- 9736082 TI - CT and MR presentation of acute hemorrhage in a spinal schwannoma. AB - We present a case of a 58-year-old woman who experienced the sudden onset of a deteriorating paraparesis after physical exertion. Pathologic specimens revealed that an intracystic hemorrhage existed within a spinal schwannoma. The radiologic presentation of the intracystic hemorrhage on computed tomography (CT) and magnetic resonance imaging (MRI) is described. PMID- 9736083 TI - Spinal intrathecal actinomycosis: a case report. AB - BACKGROUND: Actinomycosis of the central nervous system is a rare disease that most frequently forms cerebral abscesses. In the present report, we describe an extremely rare case of spinal intrathecal actinomycosis. CASE PRESENTATION: A 33 year-old man presented with high fever followed by back pain and paraparesis. Magnetic resonance imaging (MRI) with gadolinium-diethylene-triamine penta-acetic acid (Gd-DTPA) enhancement (Gd-MRI) displayed an irregularly enhanced mass lesion at the thoraco-lumbar junction that mimicked an intramedullary tumor with exophytic growth. Surgical exploration 7 months after the onset of the high fever revealed intrathecal granulation tissue with small abscess formation. Another surgical exploration was carried out 2 months after the first operation because the patient developed progressive paraparesis and showed an intrathecal ring-like enhancement that was detected with Gd-MRI. Actinomyces organisms were finally identified histologically in the surgical specimen. CONCLUSIONS: The clinical course and serial changes of Gd-MRI findings are important considerations when this rare and infectious spinal lesion is suspected. PMID- 9736085 TI - Treatment of chronic subdural hematoma by closed-system drainage without irrigation. AB - BACKGROUND: Recurrence of chronic subdural hematoma after surgery has not been eliminated. We sought to determine whether irrigation influences recurrence rate. METHODS: Patients who had undergone surgery for chronic subdural hematoma were reviewed retrospectively. RESULTS: Between 1986 and 1993, 186 cases of chronic subdural hematoma were treated surgically at Mito National Hospital. Recurrence was limited to six cases (3.2%). A closed drainage system without irrigation has been used since 1988 in 119 patients. Before 1988, 67 cases were treated with single burr hole irrigation and drainage. The rate of recurrence with the closed drainage system was 3.4% (four cases), compared with 3.0% (two cases) for irrigation and drainage. CONCLUSION: Compared with irrigation and drainage, the closed drainage system without irrigation was safe and effective, and recurrence of chronic subdural hematoma is not influenced by irrigation. PMID- 9736084 TI - Minimally invasive bedside craniotomy using a self-controlling pre-adjustable mechanical twist drill trephine. AB - BACKGROUND: Craniotomy with a mechanical twist drill is a standard, minimally invasive procedure in neurosurgery, widely used for the drainage of chronic subdural hematomas and the placement of ventricular drains. Nevertheless, the use of a standard twist drill trephine bears the risk of causing cerebral lesions. METHOD: A commercially available mechanical twist drill system has been modified by a special self-controlling drill and a pre-adjustable distance holder that limits intracerebral penetration. After initial cadaver testing, the modified trephine has been used for 65 trephinations in patients (37 chronic subdural hematomas, 21 external ventricular drains, 6 frontal hygromas, 1 tumor cyst). RESULTS: There were no complications related to the modified trephine; cerebral lesions caused by drilling too deeply or by uncontrolled penetration were safely prevented. In our series no procedure related infections occurred, and the drilling time was reduced significantly. CONCLUSION: The described modified mechanical twist drill enables fast, easy, and safe craniotomy without jeopardizing the advantages of a mechanical twist drill. Therefore, it can be recommended particularly for difficult emergency conditions. PMID- 9736086 TI - Quantitative electroencephalographic correlates of cerebral blood flow in patients with chronic subdural hematomas. AB - BACKGROUND: We have observed that the reduction in cerebral blood flow (CBF) of patients with chronic subdural hematomas is always most pronounced in the thalamus and has a linear correlation with the degree of brain deformity caused by the compression from the hematoma. When the brain is markedly displaced by a hematoma, electroencephalography (EEG) shows delta waves in the frontal region. We theorized that the flow reduction begins in the thalamus, spreading throughout the hemisphere as displacement progresses. METHODS: We measured CBF using the xenon/computed tomography (CT) method in 10 patients with chronic subdural hematomas who had hemiparesis and/or a mental disturbance and radiographic evidence of a midline shift or herniation. We correlated their flow reductions with topographic encephalograms. RESULTS: Flow reductions were more pronounced in the thalamus than in the hemisphere and cortex. Slow waves dominated over fast waves, especially in the frontopolar and frontal regions bilaterally. Flow values correlated negatively with the presence of delta and theta waves and positively with that of alpha and beta waves. On the side of the hematoma, the brain wave activity correlated significantly with thalamic flow, and the correlation was greater than with the hemispheric or cortical flow. Thalamic flow correlated more consistently with delta and alpha waves in the central, temporal, and occipital regions, and with beta waves in the frontopolar, frontal, and central regions. On the nonhematoma side, the correlation between brain wave activity and hemispheric flow was significant and greater than with cortical or thalamic flow. CONCLUSIONS: The correlation of EEGs with thalamic flow, which was maximally reduced, seems to substantiate the concept that the thalamus is primarily injured by brain distortion due to the compression from hematoma and that the remote areas are secondarily deactivated by a transneural depression ("diaschisis") originating from the dysfunctioning thalamus. Thalamic involvement seems to be the cause of the neurologic dysfunction and the abnormal EEG activity in chronic subdural hematomas. PMID- 9736087 TI - Rapid spontaneous resolution of acute subdural hematoma and HIV related cerebral atrophy: case report. AB - BACKGROUND: The natural history of a traumatic acute subdural hematoma is usually interrupted by its prompt surgical removal. Rapid spontaneous resolution within 48 hours, although infrequently reported, may be, underestimated and demonstrates a benign course of this condition. To our knowledge, this is the first case of rapid spontaneous resolution of an acute subdural hematoma in a patient with HIV encephalopathy and cerebral atrophy. METHODS AND RESULTS: This 27-year-old man, an intravenous drug user with AIDS-related complex and HIV encephalopathy, suffered an acute subdural hematoma due to head injury in a car accident. The hematoma spontaneously resolved within 12 hours, resulting in a favorable outcome with nonoperative treatment. CONCLUSIONS: AIDS related cerebral atrophy may not only have predisposed the patient to the development of an extracerebral collection, but may have also favorably influenced the spontaneous resolution of the hematoma. The mechanism of the hematoma resolution and the influence of HIV related cerebral atrophy is discussed. PMID- 9736088 TI - Reflections on the management of cerebral arteriovenous malformations. AB - BACKGROUND: The authors report their personal experience in the management of cerebral arteriovenous malformations (AVMs), using the three techniques now available: surgical resection, endovascular embolization, and radiosurgery. They review the recent literature on this topic and present their current management algorithm based on this experience. METHODS: A series of 90 patients treated for cerebral AVMs is reported (68% Grade I-III and 32% Grade IV-V, Spetzler scale). The three methods of treatment were used, either individually or in combination, based on the size and the location of the malformation. The first intervention was surgical resection in 26% of cases, endovascular embolization in 57%, and radiosurgery in 17%. Surgery and embolization were followed by another technique in some cases and eventually single modality treatment was used in 58% of cases (surgical resection 21%, endovascular embolization 20%, radiosurgery 17%) and multimodality treatment in 42% (embolization + resection, 21%; embolization + radiosurgery, 17%; resection + radiosurgery, 4%). Embolization was used as reductive therapy in 38% of the overall series (65% of all embolized patients), and was followed by surgery in 56% of cases or by radiosurgery in 44%. Angiography was used to assess the cure rates. RESULTS: The following cure rates were obtained, when each technique was used as a first treatment: surgical resection, 82%; embolization, 6%; and radiosurgery, 83% (2-year angiographic follow-up). After combined treatment, embolization and resection resulted in a 100% cure rate, embolization and radiosurgery produced a 90% cure rate. The clinical outcome was evaluated in terms of deterioration attributable to treatment. Seventy-one percent of patients had no complication, minor complications were observed in 18%, and severe complications in 11%. Treatment mortality was 3%. All deaths were attributable to hemorrhage during the embolization procedure. CONCLUSIONS: In this management algorithm, AVMs submitted directly to surgery or to radiosurgery were considered "good risk" malformations, and the outcome for these cases was good in terms of clinical result and cure rate. AVMs submitted first to endovascular embolization were considered "poor risk" malformations, including a majority of Spetzler Grade IV-V lesions. Not surprisingly, the majority of severe complications occured in this group during embolization. Thus, the major risk of the treatment of AVMs has now shifted from surgery to endovascular techniques. Endovascular embolization as sole treatment gave a low rate of complete occlusion, but proved to be very useful as a reductive therapy, in preparation for further surgery or radiosurgery. Partial embolization permitted high rates of complete cure in difficult AVMs. Embolization should be used to the maximum extent possible as a reductive technique, despite the risks of the procedure. Because of its risks however, this technique of reductive embolization should be used only if absolutely necessary to allow the complete cure of the malformation. Thus, the use of embolization should be considered very cautiously in small malformations as well as in very large and complex AVMs in which partial embolization will not be sufficient to allow complete cure with either endovascular or surgical techniques. PMID- 9736089 TI - Intracarotid pressure measurements in the evaluation of a computer model of the cerebral circulation. AB - BACKGROUND: It is difficult to predict which patients will tolerate occlusion of the internal carotid artery. This difficulty arises primarily because of uncertainties in the prediction of the adequacy of collateral circulation. Because of these uncertainties, balloon test occlusion and other methods have been developed to determine a priori the safety of carotid occlusion. However, all the methods are associated with significant false-positive and false-negative rates, as well as other neurologic complications. Because of these problems, more accurate and less invasive methods for predicting tolerance of carotid occlusion are needed. METHODS: In this report, we present the initial clinical evaluation of a new method for assessing the collateral circulation aided by a mathematical model of the cerebral vasculature. Data from the angiograms of 14 patients who underwent carotid endarterectomy were used to create individualized simulations of their cerebral circulations. As a test of the accuracy of the simulations, we compared values of the intracarotid stump pressures predicted by the model to those measured at surgery during the period of carotid occlusion. RESULTS: The pressure predictions of the model correlated well with those measured at surgery. Linear regression analysis of measured versus predicted values yielded a line with slope 1.05. The line with slope 1.00, which denotes perfect agreement between predictions and measurements, is within the 95% confidence interval of the slope determined from the regression analysis. CONCLUSIONS: Mathematical models of the cerebrovascular circulation can provide good predictions of intravascular pressure in the collateral circulation, and may provide accurate predictions of the flow as well. The present study reveals several areas that need further development, such as the models of the microvasculature, measurement of the arterial dimensions from angiograms, and consideration of other collateral sources such as the leptomeningeal and retrograde ophthalmic sources of flow. Incorporation of these improvements may lead to a clinically useful, noninvasive assessment of the state of the cerebrovascular collateral circulation in the individual patient. PMID- 9736093 TI - Proposal concerning neurosurgical residency financing. PMID- 9736092 TI - Cystic mass of the conus in a child. PMID- 9736091 TI - Combined transpetrosal and fronto-orbito-zygomatic approach to a giant skull based meningioma: a case report. AB - BACKGROUND: Recently, various surgical approaches to skull base lesions have been developed. Skillful use of the combination of two standard approaches make possible the removal of large brain lesions, which conventionally had been considered inoperative. In this study, we present a case of a giant meningioma located in the cerebellopontine angle and middle cranial fossa. A near total resection was achieved using a combined transpetrosal and fronto-orbito-zygomatic approach. CASE REPORT: A 15-year-old boy presented with a meningioma that caused a left hearing loss, dysarthria, and cerebellar ataxia. Preoperative magnetic resonance imaging revealed a giant meningioma located in the right cerebellopontine angle, middle fossa, and cavernous sinus. The patient underwent a near total resection of the tumor through a combined transpetrosal and fronto orbito-zygomatic approach. He experienced a marked improvement postoperatively and entered high school the following year. CONCLUSIONS: An approach from several angles was necessary for the giant skull based tumor presented here. A combination approach was selected for obtaining a wide operative field with minimal brain compression during resection of neoplasm. According to the individual features of each case, selection of the operative approach, decisions regarding the extent of excision, and postoperative treatment regimens should be adequately planned in the preoperative period. PMID- 9736090 TI - Tizanidine is an effective agent in the prevention of focal cerebral ischemia in rats: an experimental study. AB - BACKGROUND: Focal cerebral ischemia secondary to cerebral vessel occlusion is still an important cause of mortality and morbidity. Excitatory neurotransmitters are gathered in the extracellular space during ischemia and initiate or stimulate a series of pathophysiological biochemical processes and consequently lead to neuronal death. Tizanidine (Sandoz compound DS 103-282, 5-chloro4,2 (2-imidazolin 2-yl-amino)-2,1,3-benzothiazol hydrochloride) is a selective alpha 2 adrenoreceptor agonist which shows its effect by stimulating presynaptic alpha 2 adrenoreceptors in central ASPergic and GLUergic system by inhibiting aspartic acid and glutamic acid release. In this study, the effect of Tizanidine on reversible focal cerebral ischemia was evaluated. METHODS: Cerebral blood flow to the left hemisphere of adult Sprague-Dawley rats (n=48) was temporarily interrupted by middle cerebral artery and bilateral common carotid artery occlusion for 3 hours in eight rats of each group. Tizanidine was given to each group of rats intraperitoneally before the ischemic insult, 2 hours after ischemia, right after the reperfusion, 2 h after reperfusion, and 4 hours after reperfusion; the animals survived for 24 hours after the reperfusion. After killing and triphenyltetrasoliumchloride staining of brain slices, infarction volumes and ratios of the brains were calculated and the results were compared with those of the control group. RESULTS: Infarction volumes and infarction ratios of the Tizanidine group 1/2 hours before ischemia (143.7+/-6.34 mm3 and 10.1+/-0.43%) and the Tizanidine group 2 hours after ischemia (145.6+/-6.32 mm3 and 10.3+/-0.43%) were found to be significantly lower in favor of the Tizanidine groups when compared with those of the control group (173.9+/-6.38 mm3 and 12,4+/ 0.41%). Tizanidine is not effective if used just after reperfusion or later. CONCLUSION: This study shows that Tizanidine pretreatment before the ischemic insult and the administration of the drug within the 2 hours after ischemia reduces ischemic damage significantly. Therefore, this drug can be used as a protective and therapeutic agent in ischemic diseases. PMID- 9736094 TI - Don't sell the farm. PMID- 9736095 TI - Management of acute subdural hematoma. PMID- 9736096 TI - Acute subdural hematoma of arterial origin. PMID- 9736097 TI - Prevalence rates of hearing impairment and comorbid conditions in older people: the Veneto Study. AB - OBJECTIVES: To investigate the prevalence rate of hearing impairment, assessed by both the Sanders' questionnaire and the speech audiometry test, and its association with health-related factors in the older population of the Veneto region of Italy. DESIGN: A cross-sectional survey. SETTING: A community-based population. PARTICIPANTS: 2398 noninstitutionalized individuals aged 65 years and older residing in the Veneto region of Italy. MEASUREMENTS: Prevalence rates of hearing impairment and odds ratios for its association with potential risk factors. MAIN RESULTS: The prevalence of self-reported hearing impairment at home was 8.1% in men and 7.4% in women, and in a social environment it was 11.1% and 9.3%, respectively. Women were less likely to report hearing difficulties in both environments, and increased risks were found for depression, age, and poor self rated health. Participants with diabetes or cognitive impairment had increased odds only at home, in contrast to people with a low education level, who had increased odds only in a social environment. The prevalence assessed by speech audiometry was 19% in both sexes. Increased age, diabetes, and poor self-rated health were associated with impaired speech intelligibility, cognitive impairment was associated with 4-fold increased odds among past users of alcohol, and men with a low education level were about three times as likely as others to have hearing impairment. CONCLUSIONS: Speech audiometry testing detected a higher prevalence of hearing impairment than use of a self-reported questionnaire and was associated with poor self-rated health, history of diabetes, and cognitive impairment among past users of alcohol and among men with low levels of education. The association between hearing deficit and depressive symptomatology was confirmed only with self-reported hearing impairment. PMID- 9736098 TI - Low prevalence of hearing aid use among older adults with hearing loss: the Epidemiology of Hearing Loss Study. AB - OBJECTIVES: To measure the prevalence of hearing aid use among older adults with hearing loss and to identify factors associated with those currently using hearing aids. DESIGN: Population-based cohort study. SETTING: The south-central Wisconsin community of Beaver Dam. PARTICIPANTS: A total of 1629 adults, aged 48 to 92 years, who have hearing loss and are participating in the Epidemiology of Hearing Loss Study and the Beaver Dam Eye Study. MEASUREMENTS: A hearing-related risk factor and medical history questionnaire, the Hearing Handicap Inventory for the Elderly (screening version), screening tympanometry, pure-tone air- and bone conduction audiometry, and word recognition tests were administered by trained examiners using standard protocols. RESULTS: The prevalence of current hearing aid use among those with a hearing loss (pure-tone average > 25 decibels hearing level over 500, 1000, 2000, and 4000 Hertz, worse ear) was 14.6%. The prevalence was 55% in a subset of the most severely affected participants. In univariate analyses, current hearing aid use was associated with age, severity of loss, word recognition scores, self-reported hearing loss, self-perceived hearing handicap, and history of noise exposure. Factors associated with current hearing aid use in multivariate logistic regression models were age, severity of loss, education, word recognition scores, Hearing Handicap Inventory for the Elderly (screening version) score, and self-report of a hearing loss. CONCLUSIONS: Few older adults with hearing loss are currently utilizing hearing aids. Improved screening and intervention programs to identify older adults who would benefit from amplification are needed to improve hearing-related quality of life for this large segment of the population. PMID- 9736099 TI - Treatment of pain in cognitively impaired compared with cognitively intact older patients with hip-fracture. AB - OBJECTIVE: To compare the experience of pain and treatment of pain in cognitively impaired and cognitively intact older adults after surgical repair of a hip fracture. DESIGN: Prospective comparative survey design. PARTICIPANTS: A convenience sample of 88 hip fracture patients (53 cognitively impaired, 35 cognitively intact) from three Midwestern urban hospital orthopedic units was interviewed between days 2 and 5 postoperatively. Subjects whose Folstein Mini Mental State Exam (MMSE) score was less than or equal to 23 were categorized as impaired. RESULTS: Pain report and intensity did not differ significantly between the two groups. One-third of the subjects in both groups rated pain as severe or worse. Cognitively impaired subjects scored significantly higher on the Checklist of Nonverbal Pain Indicators observed with movement (CNPI-m) than did cognitively intact subjects. Cognitively impaired subjects received significantly less opioid analgesics than cognitively intact subjects in the first and second 48 hours postoperatively. Both groups received less than 25% of the mean prescribed amount of opioid analgesics. Age, MMSE, and CNPI-m score accounted for 27% of the variance in the amount of opioid analgesic administered in the first 48 hours postoperatively. CONCLUSIONS: Pain is treated poorly in older postoperative patients. Cognitive impairment and age strongly influence the amount of analgesic nurses administer to older patients after surgical repair of hip fracture. Provision for patient comfort is a fundamental ethical obligation of healthcare providers. Clinicians need to pursue this goal more aggressively, especially for cognitively impaired, postoperative older adults. PMID- 9736100 TI - The effectiveness of day hospital care on home care patients. AB - OBJECTIVE: To study the effectiveness of rehabilitative and medically oriented day hospital care on community-based long-term care patients. DESIGN: A randomized, controlled trial. SETTING AND PARTICIPANTS: 177 patients on home-care in a rural area were randomized into two groups. Patients in one group were offered a 2-month period of rehabilitation and medical care in a recently opened day hospital, and in the other group patients were offered treatment, as before, in home care. Both groups were examined at the beginning and at 2, 5 and 12 months. INTERVENTION: Rehabilitative and medically oriented day hospital care. OUTCOME MEASURES: Use of health services, physical functioning measured by the Katz ADL Index, subjective health, symptoms, and satisfaction with care. RESULTS: The groups used hospitals (excluding the day hospital treatment) equally during the follow-up year. The treatment group had significantly more specialist consultations than did the control group. There were no clinically significant differences in the changes in the Katz ADL Index although more changes were found in the treatment group. The number of symptoms was reduced significantly in the treatment group, whereas the number of symptoms remained unchanged in the control group. The patients' views of their own health improved in the treatment group. CONCLUSION: Day hospital care affects the quality of life of older people, but it does not reduce the use of other health services, nor does it clinically significantly improve the physical functioning of older people. PMID- 9736101 TI - The experience of living-dying in a nursing home: self-reports of black and white older adults. AB - PURPOSE: The purpose of this study was to describe and compare the experiences, needs, priorities, and concerns reported by black and white nursing home residents during the living-dying interval. The living-dying interval is defined as the time between the knowledge of one's impending death and death itself. DESIGN: This qualitative study was part of a larger ethnographic project. Residents participated in from one to four individual, in-depth, semi-structured, audiotaped interviews. SETTING: Residents lived in two large county-financed nursing homes that have historically provided care to indigent black and white older adults. PARTICIPANTS: Purposive sampling was used to identify eight black and five white residents with terminal cancer diagnoses who could serve as rich resources about the experience of living-dying in a nursing home. MEASURES: Residents were asked open-ended questions about how things have been and what would make things better; what comforts them and would make them more comfortable; what dying means to them; and what things are important for nursing staff to know. RESULTS: Verbatim transcripts of the interviews were coded using QRS NUD-IST software. Codes were placed in categories, categories were reviewed for common and different concepts, themes, and patterns, and a conceptual model was developed. The model identified six care needs: (1) day-to-day living; (2) inadequate pain relief for black residents; (3) difficulty chewing and swallowing; (4) importance of religious activities; (5) giving care to others; and (6) appreciation of respectful and prompt care. Residents validated all components of the conceptual model. CONCLUSION: Black and white terminally ill residents focused on the quality of living rather than on dying, and black residents may be undertreated for pain. Important care needs for pain and religion are not routinely addressed by the Minimum Data Set (MDS) and Resident Assessment Protocol (RAP) triggers. PMID- 9736102 TI - A prospective study of the efficacy of the physician order form for life sustaining treatment. AB - OBJECTIVES: The Physician Orders for Life-Sustaining Treatment (POLST), a comprehensive, one-page order form, was developed to convey preferences for life sustaining treatments during transfer from one care site to another. This study examined the extent to which the POLST form ensured that nursing home residents' wishes were honored for Do Not Resuscitate (DNR) and requests for transfer only if comfort measures fail. DESIGN: The study used chart record data to follow prospectively a sample of nursing home residents with the POLST. SETTING: Eight geographically diverse, long-term, adult-care facilities in Oregon in which the POLST was in use. PARTICIPANTS: Nursing home residents (n = 180), who had a POLST recording DNR designation and who indicated a desire for transfer only if comfort measures failed, were followed for 1 year. MEASUREMENTS: For all subjects: treatment and disposition after significant health status changes; orders for narcotics and for provision or limitation of aggressive interventions. For hospitalized subjects: diagnosis, medical interventions, and DNR orders. For those who died: cause and location of death, life-sustaining treatments attempted, and comfort measures provided. RESULTS: No study subject received CPR, ICU care, or ventilator support, and only 2% were hospitalized to extend life. Of the 38 subjects who died during the study year, 63% had an order for narcotics, and only two (5%) died in an acute care hospital. A total of 24 subjects (13%) were hospitalized during the year. Hospitalized subjects' mean length of stay was 4.9 days, and the mean rate of hospitalizations for all subjects was 174 per 1000 resident years. In 85% of all hospitalizations, patients were transferred because the nursing home could not control suffering. In 15% of hospitalizations (n = 4), the transfer was to extend life, overriding POLST orders. CONCLUSIONS: POLST orders regarding CPR in nursing home residents in this study were universally respected. Study subjects received remarkably high levels of comfort care and low rates of transfer for aggressive life-extending treatments. PMID- 9736103 TI - Elder-proxy agreement concerning the functional status and medical history of the older person: the impact of caregiver burden and depressive symptomatology. AB - OBJECTIVES: To examine the influence of caregiver burden and depressive symptomatology on elder-proxy response concordance regarding the older person's functional status and medical history. DESIGN: Cross-sectional study via telephone interviews. SETTING: Community-dwelling older people and caregivers in North Carolina. PARTICIPANTS: 340 matched pairs of frail persons aged 65 and older and their respective caregivers. MEASUREMENTS: Multidimensional Functional Assessment: The OARS methodology RESULTS: Percent agreement on the ADL items ranged from 97.6% on personal hygiene to 99.7% for toileting, with moderate kappa coefficients. IADL percent agreement ranged from 71.5 to 93.7%, with fair to moderate kappa coefficients. Agreement among the medical history items ranged from 76.3 to 98.5% (kappa = .138-.831). Response bias for the IADL composite measure is influenced marginally by caregiver burden (F[259] = 1.751, P = .098). Five of the individual IADL bias items are influenced significantly by burden, such that an increase in burden results in a greater likelihood that the caregiver will overstate disability compared with the rating by the older person. Response bias on the ADL scale was increased among persons who experienced more caregiver burden (OR = 1.096, 95% CI = 1.000, 1.192) and those who spent more hours providing care (OR = 1.012, 95% CI = 1.001, 1.024). Additionally, black caregivers were more likely than white caregivers to disagree with the older people on the ADL scale (OR = 2.73, 95% CI = 1.642, 3.809). A composite of the medical history items is influenced by the relationship of the caregiver to the older person; bias is more likely among adult children ((F[227] = 1.56, P = .081). CONCLUSION: Elder-proxy concordance is highest among ADL items, followed by medical history items and IADL items. Caregiver depressive symptomotology had no significant impact on elder-proxy response concordance on any of the three outcomes of interest: IADL and medical history bias and ADL disagreement. However, caregiver burden was marginally predictive of bias on the total ADL and IADL scales. Additionally, increased burden was significantly predictive of bias on five of the seven individual items of the IADL scale, suggesting that the more burden a caregiver feels, the greater likelihood that s/he will overstate the older person's disability compared with self-report. These findings suggest that clinicians and researchers who use proxy reports to determine treatment regimens and complete data collection efforts may do so with confidence on ADL individual items and medical history items when the older person's frailty is marginal. However, caregiver burden may result in misleading representation of the older person's functional status, specifically in regard to IADL items. PMID- 9736104 TI - Outbreak of pneumonia in a long-term care facility: antecedent human parainfluenza virus 1 infection may predispose to bacterial pneumonia. AB - OBJECTIVES: To determine the causes of an outbreak of lobar pneumonia. DESIGN: Matched (1:2) case-control study. SETTING: A 70-bed chronic care facility for older people. PARTICIPANTS: Residents of the facility. RESULTS: Ten residents developed pneumonia over a 10-day period. Two residents died. One case-patient had Streptococcus pneumoniae bacteremia; another had polymerase chain reaction evidence of S. pneumoniae infection. No other etiologic agent was identified. Only four of 10 case-patients had received routine diagnostic cultures of blood or sputum before the administration of antibiotics. Symptoms of upper respiratory illness (URI) among residents before the pneumonia outbreak corresponded with elevation of antibodies to human parainfluenza virus 1 (HPIV1). In a matched case control study, six of 10 case-patients, compared with five of 20 controls, had symptoms of URI during the preceding month (matched odds ratio (MOR) = 4.5, 95% CI = 0.8-33). Nine case-patients had serum available, and five of these had both serologic evidence of recent HPIV1 infection and recent URI, compared with two of 18 controls (MOR = 9.0, 95% CI = 1.2-208). Only three residents had documentation of pneumococcal vaccination. CONCLUSIONS: Noninfluenza viral infections may play a role in the pathogenesis of some bacterial pneumonias. S. pneumoniae was the cause of at least two pneumonias; lack of preantibiotic cultures may have interfered with isolation of S. pneumoniae in others. Recent HPIV1 infection was epidemiologically linked to subsequently developing pneumonia. Spread of HPIV1 in the facility may have contributed to increased susceptibility to S. pneumoniae and, potentially, to other bacterial pathogens. PMID- 9736105 TI - Improving the utility of urine flow rate to exclude outlet obstruction in men with voiding symptoms. AB - OBJECTIVES: Many older men with voiding symptoms do not have bladder outlet obstruction (BOO) but have conditions amenable to medical treatment. If primary care providers could reliably exclude men who have BOO, then they could initiate appropriate evaluation and treatment in a substantial proportion of the remainder. Urine flow rate, although widely used, is insufficiently sensitive to exclude BOO reliably. We investigated whether the decline in flow rate with age could be utilized to improve the utility of flow rate for excluding BOO in symptomatic men, especially when combined with knowledge of the patient's postvoiding residual volume (PVR). DESIGN: Prospective study using two patient cohorts. SETTINGS: A Veterans Affairs urology clinic serving community-dwelling and institutionalized healthy older men. PARTICIPANTS: 111 men with voiding symptoms (mean age 72.4 +/- 9.2 years). MEASUREMENTS: Maximum urine flow rate, measured with standard flowmeter, and PVR, measured by catheterization. BOO was determined by multichannel videourodynamic testing. RESULTS: The sensitivity of flowrate for BOO increased significantly with age (P = .0001) and did not appear to be confounded by comorbid conditions. An algorithm incorporating age, flow rate, and PVR had a sensitivity of 90%, specificity of 43%, and accuracy of 74% in screening for BOO. The algorithm's sensitivity was better than that of flow rate alone (55%); its sensitivity was also similar to a "refer all" strategy (100%) but had improved specificity (41% vs 0%). CONCLUSIONS: Flow rate alone is insufficiently sensitive as a screening test to exclude BOO, but a simple diagnostic algorithm using age, flowrate, and PVR was more sensitive and accurate. This algorithm allows primary care evaluation and initial management of men with voiding symptoms while potentially reducing unnecessary referrals and costs. PMID- 9736106 TI - Oral cobalamin for pernicious anemia: back from the verge of extinction. AB - BACKGROUND: High dose oral cobalamin therapy was shown to be effective for pernicious anemia and other cobalamin deficiency states 30 years ago, and physicians and patients state that they would find oral therapy useful, but a survey conducted in 1989 found that physicians were generally unaware of it. OBJECTIVE: To assess physician awareness and use of oral cobalamin since 1989. DESIGN, SETTING, AND PARTICIPANTS: Minneapolis area internists not listed as having subspecialties or academic business addresses were surveyed in 1989 and in 1996. MEASUREMENTS AND RESULTS: There were 245 responses to the 1989 survey and 223 responses to the 1996 survey for response rates of 68% and 69%, respectively. The percentage of internists who ever used oral cobalamin to treat pernicious anemia increased from 0 in 1989 to 19% in 1996 (P < .001). The percentage who were aware of an effective oral cobalamin preparation for treating cobalamin deficiency states also increased significantly from 4 to 29% (P < .001). The percentage of internists who agreed with the incorrect view that sufficient quantities of cobalamin cannot be absorbed when given orally declined from 91% in 1989 to 71% in 1996 (P < .001). CONCLUSION: Minneapolis internists' awareness and use of oral cobalamin treatment for pernicious anemia increased substantially between 1989 and 1996, but the majority of internists remained unaware of this treatment option. PMID- 9736107 TI - Underutilization of measurement of serum low-density lipoprotein cholesterol levels and of lipid-lowering therapy in older patients with manifest atherosclerotic disease. AB - OBJECTIVE: To investigate the prevalence of measurement of serum lipids and of utilizing lipid-lowering therapy, when appropriate, in older persons without contraindications to lipid-lowering drugs and with myocardial infarction (MI), stroke, peripheral arterial disease (PAD), and no coronary artery disease (CAD), stroke, or PAD. DESIGN: A retrospective analysis of charts of all older patients seen from January 1, 1997, through October 15, 1997, was performed to investigate the prevalence of measurement of serum lipids and utilization of lipid-lowering therapy, when appropriate, in older persons without contraindications to lipid lowering drugs and with MI, stroke, PAD, and no CAD, stroke, or PAD. Patients with life-limiting comorbidities were not included in the study. SETTING: An academic, hospital-based geriatrics practice staffed by fellows in a geriatrics training program and full-time faculty geriatricians. PARTICIPANTS: A total of 373 men and 1119 women, mean age 80 +/- 8 years (range 59 to 103 years), were included in the study. MEASUREMENTS AND MAIN RESULTS: Serum low-density lipoprotein (LDL) cholesterol was measured in 201 of 391 patients (51%) with MI, in 78 of 187 patients (42%) with stroke, in 58 of 115 patients (50%) with PAD, and in 396 of 926 patients (43%) with no CAD, stroke, or PAD. In patients with elevated serum LDL cholesterol levels, lipid-lowering drug therapy was given to eight of 15 patients (53%) <70 years of age with MI, to 34 of 63 patients (54%) 70 to 80 years of age with MI, and to 38 of 81 patients (47%) >80 years of age with MI (P not significant); to three of seven patients (43%) <70 years of age with stroke, to 12 of 26 patients (46%) 70 to 80 years of age with stroke, and to 13 of 32 patients (41%) >80 years of age with stroke (P not significant); to two of four patients (50%) <70 years of age with PAD, to seven of 17 patients (41%) 70 to 80 years of age with PAD, and to 10 of 25 patients (40%) >80 years of age with PAD (P not significant); and to six of 15 patients (40%) <70 years of age with no CAD, stroke, or PAD, to 26 of 70 patients (37%) 70 to 80 years of age with no CAD, stroke or PAD, and to 14 of 47 patients (30%) >80 years of age with no CAD, stroke, or PAD (P not significant). None of the other patients with MI, stroke, PAD, or no CAD, stroke, or PAD was treated with diet or lipid-lowering drugs. CONCLUSIONS: Measurement of serum LDL cholesterol and of appropriate use of lipid-lowering drugs and diet in older patients with MI, stroke, PAD, and no CAD, stroke, or PAD is underutilized in an academic, hospital-based geriatrics practice. PMID- 9736108 TI - Compliance with PASARR recommendations for Medicaid recipients in nursing homes. Preadmission Screening and Annual Resident Review. AB - OBJECTIVE: To determine the rate of compliance with placement and mental health recommendations of the preadmission screening and annual resident review (PASARR) program. DESIGN: A retrospective observational study using PASARR screening forms to identify recommendations of inpatient psychiatric care for people needing more care than nursing homes provide, alternate disposition for individuals needing less than nursing home level care, and recommendations for new mental health services when needed but not provided. Service use as indicated in Medicaid billing data during the following year was used to determine receipt of service. PARTICIPANTS: All Washington state Medicaid recipients screened from 1992 through 1993. MAIN OUTCOME MEASUREMENTS: Compliance rates for placement and service recommendations. RESULTS: Inpatient psychiatric care was recommended for four of the 523 Medicaid recipients (0.8%), all of whom received it. Screeners recommended alternate dispositions in 131 (25%) subjects, and compliance occurred in 29% of these. Recommendations for new services were made in 310 (59%) cases. Compliance rates averaged 35%, ranging from 73% for medication recommendations to 7% for consultation. Depressed individuals were less likely to receive recommended services. CONCLUSIONS: Many individuals needed additional mental health services but did not receive them, and a significant minority of patients could have been given an alternate disposition but rarely were. PMID- 9736110 TI - Should governments force people to save for retirement? PMID- 9736109 TI - An evaluation of antiepileptic drug therapy in nursing facilities. AB - OBJECTIVES: To describe the prescribing and use of antiepileptic drug (AED) therapy in nursing facility residents. DESIGN: A retrospective, multicenter drug use evaluation. SETTING: A total of 85 nursing facilities (average size, 119 beds) in five states. PARTICIPANTS: 1132 residents of the total 10,168 residents screened were prescribed at least one AED. MEASURES: Demographic information, primary indication for AED, comorbid conditions, prescribing physician's specialty, concomitant medications, and AED dosage regimen information were collected. Laboratory tests obtained in the most recent 6 months and seizure occurrence and seizure-related diagnostic assessments made in the most recent 3 months were also recorded. RESULTS: Of 1132 residents receiving AED therapy, 892 (78.8%) were prescribed AED therapy for a seizure-related diagnosis although 86% of seizure types were unspecified. Another 215 residents (19.0%) were prescribed AEDs for nonseizure diagnoses, and 25 (2.2%) had no indication for AED therapy. AEDs most frequently prescribed were phenytoin (56.8%), carbamazepine (23.0%), phenobarbital (15.6%), and valproic acid (13.1%). For residents with a seizure diagnosis, the most frequently prescribed monotherapy agents were phenytoin (52.0%), carbamazepine (12.2%), and phenobarbitol (7.1%). Almost 25% of residents with a seizure diagnosis took a combination of AEDs; more than 50% of all combinations included phenobarbital. About 9% of residents with a seizure diagnosis had one or more documented seizures during a 3-month review period. CONCLUSION: Among the substantial percentage of residents treated with AEDs, the lack of diagnosis of seizure type has serious implications for the choice of AED therapy. Opportunities exist for prescribing physicians, consultant pharmacists, and nursing staff to improve the medical management of nursing facility residents with seizures and of others receiving AEDs. PMID- 9736111 TI - Shoulder pain in older people. AB - Shoulder pain is encountered commonly in older people. Most of the conditions are amenable to nonoperative treatment, but a clear understanding of the anatomy and those conditions encountered most commonly is essential. A careful history and physical examination, as well as limited diagnostic tests, point to a clear diagnosis in the majority of patients. Most often, conservative measures are effective. Steroid injections are often helpful for both diagnostic and therapeutic reasons. A small percentage of patients do not respond to conservative management and require operative intervention. PMID- 9736112 TI - Managed primary care of nursing home residents. AB - OBJECTIVE: To measure the rates of hospital use and mortality of nursing homes residents who received their primary care from nurse practitioner-physician teams. DESIGN: A cohort study. SETTING: Thirty nursing homes in Southern California. PATIENTS: Older, long-term residents of nursing homes enrolled in a Medicare HMO (n = 307). INTERVENTION: Primary care by an accessible interdisciplinary team supported by clinical guidelines, continuous quality improvement techniques, and increased availability of clinical services at the nursing homes. RESULTS: The residents (mean age 83.5 years, 69.0% women) had a high prevalence of dementia (83.5%) and functional disability (87.2% were dependent in two or more activities of daily living). About half (50.8%) expressed a preference for "no hospitalization and no resuscitation." Compared with other nursing homes populations, this cohort experienced a lower annual rate of hospital use (518 days/1000 residents) and a similar rate of mortality (23.8%). CONCLUSIONS: Integration of the efforts of physicians, nurse practitioners, and nursing home staff can lead to low rates of hospital use by nursing home residents. The effects on residents' quality of life and mortality require further study. PMID- 9736113 TI - Management of the older patient with acute myocardial infarction: difference in clinical presentations between older and younger patients. AB - The majority of persons sustaining acute myocardial infarction are older, and in these older persons morbidity and mortality are high. Clinical presentations and characteristics are significantly different between older and younger infarction patients. Older infarction patients are more likely to be female and to have a history of heart failure, but they are less likely to have a family history of myocardial infarction, elevated cholesterol, or to smoke. Older patients will frequently have unrecognized or silent myocardial infarctions or, when present, symptoms will be atypical. Instead of chest pain, older patients may have shortness of breath or neurological symptoms, such as confusion. Also, older infarction patients will delay longer in seeking medical assistance after onset of symptoms, and often will not demonstrate ST elevation or Q waves on their electrocardiograms. Not infrequently, older infarction patients will demonstrate major complications such as heart failure or right ventricular infarction on hospital admission, and their presenting complaints will reflect these complications. Because of these atypical presentations and the wide variability of symptoms, physicians must be highly suspicious of the presence of an acute myocardial infarction in older patients who have an unexplained acute change in their physical condition. PMID- 9736114 TI - Benign prostatic hyperplasia: diagnosis and treatment. AHCPR. Agency for Health Care Policy and Research. PMID- 9736115 TI - Better use of hearing aids in hearing-impaired adults. PMID- 9736116 TI - Looking beyond the "form" to complex interventions needed to improve end-of-life care. PMID- 9736117 TI - Care of the living, care of the dying: reconceptualizing nursing home care. PMID- 9736118 TI - Impaired one-leg balance as a cause of falls. PMID- 9736120 TI - No decline in cognitive function in older patients two months after a fracture of the extremities. PMID- 9736119 TI - Geropsychiatric restraint use. PMID- 9736121 TI - Benzodiazepine users aged 85 and older fall more often. PMID- 9736122 TI - Hemodynamic cerebral infarction triggered by excessive blood pressure reduction in hypertensive emergencies. PMID- 9736123 TI - Patient risk or physician practice: what factors drive resource use in unstable angina? PMID- 9736124 TI - Coronary angiography before myocardial infarction: can the culprit site be prospectively recognized? PMID- 9736125 TI - Is 6-minute walk test of value in congestive heart failure? PMID- 9736126 TI - Clinical predictors easily obtained at presentation predict resource utilization in unstable angina. AB - OBJECTIVE: To determine if a risk prediction model for patients with unstable angina would predict resource utilization. METHODS AND RESULTS: Four hundred sixty-five consecutive patients admitted for unstable angina to a tertiary care university-based medical center were prospectively evaluated from June 1, 1992, to June 30, 1995. The proportion of patients receiving coronary angiography, coronary angioplasty, and coronary artery bypass grafting were analyzed according to four risk groups on the basis of a previously published model: Group 1, <2% risk of major complication; Group 2, 2.1% to 5% risk; Group 3, 5.1 % to 15% risk; and Group 4, >15.1 % risk. Hospital length of stay and estimated cost of hospitalization based on DRG and specific payer ratio of cost-to-charge were also compared between groups. Multiple linear regression analysis was used to determine the influence of estimated risk and procedures on hospital costs. The four groups were well matched for gender, hypertension, tobacco history, and previous percutaneous transluminal coronary angioplasty and myocardial infarction. Group 4 had a higher incidence of previous coronary bypass grafting (35% vs 10%, p=0.001) and triple vessel or left main coronary artery disease compared with Group 1 (44% vs 13%, p=0.041). Group 4 patients were more likely to be admitted to the coronary care unit compared with Group 2 or Group 1 patients (80% vs Group 1: 51% [p= 0.001]; and vs Group 2: 53% [p=0.001]), more likely to receive heparin (87% vs 71%, p=0.007), and more likely to receive a beta-blocker or calcium channel blocker (89% vs 74%, p=0.008) than Group 1. Coronary angioplasty rates were similar for all groups, but Group 4 patients were more likely to receive coronary bypass grafting than Group 2 or Group 1 (27% vs Group 2: 12%, p=0.004 and vs Group 1: 8%, p=0.002). Hospital length of stay was highest in Group 4 and lowest for Group 1. Average hospital costs were significantly less in Group 3 than in Group 4, but higher than in Group 1. Multivariate analysis determined a dependency of costs on risk group with Group 2 having costs 31.4% (95% CI=9.8 to 57.2), Group 3 46.7% (24, 3 to 73.1), and Group 4 75% (46.9 to 110.7) higher than Group 1. The use of procedures also significantly increased costs, with PTCA-treated patients having a 44.9% (26.7 to 65.7) increase in costs compared with medically treated patients, and surgically treated patients having a 204.7% increase in costs. CONCLUSION: Resource utilization as assessed by the use of revascularization procedures, length of stay, and hospital costs are influenced by patient acuity estimated from a prediction model on the basis of estimated risk of cardiac complications. The model exerts independent influence on cost even after adjustment for various procedures. The use of revascularization procedures, especially coronary artery surgery, remains a large determinant of hospital cost. PMID- 9736127 TI - Angiographic characteristics of infarct-related and non-infarct-related stenoses in patients in whom stable angina progressed to acute myocardial infarction. AB - BACKGROUND: In patients with coronary artery disease, angiographic and postmortem studies have shown that coronary stenoses in infarct-related arteries often have complex morphology. It is not known whether in patients with multivessel disease stenosis morphology in non-infarct-related arteries is different from those of the infarct-related arteries. METHODS AND RESULTS: In 24 consecutive patients we examined the angiographic characteristics of both the infarct-related stenoses and non-infarct-related stenoses before and after spontaneous acute myocardial infarction, by visual inspection and computerized edge detection of coronary angiograms. Before myocardial infarction, the severity of the infarct-related stenoses was <50% in 14 patients and > or =50% in 10 patients (p=not significant) and of non-infarct-related stenoses was <50% in 16 and > or=50% in 13. A significantly greater proportion of infarct-related stenoses with severity > or =50% progressed to non-Q-wave than to Q-wave myocardial infarction (71% vs 50%, p < 0.05). Before myocardial infarction, the percentage of concentric, eccentric, and irregular infarct-related stenoses was 8%, 13%, and 50%, respectively, whereas in the non-infarct-related stenoses it was 62%, 17%, and 21%, respectively (p < 0.01). A similar proportion of irregular morphology progressed to Q-wave or non-Q-wave myocardial infarction. CONCLUSIONS: In patients with stable angina who had acute myocardial infarction develop, the infarct-related and non-infarct-related stenoses on average are similar in severity but different in morphology. Nonsevere stenoses more frequently progress to Q-wave than to non Q-wave myocardial infarction. PMID- 9736128 TI - Evidence of endothelial dysfunction of epicardial coronary arteries in patients with immunohistochemically proven myocarditis. AB - BACKGROUND: Recent reports indicate that myocarditis can be associated with acute myocardial ischemia and even myocardial infarction in patients with normal arteriograms. We therefore tested the hypothesis that patients with biopsy-proven myocarditis have endothelial dysfunction despite angiographically smooth epicardial coronary arteries. METHODS AND RESULTS: Graded concentrations of the endothelium-dependent vasodilator acetylcholine (10(-6) to 10(-4) mol/L) and for comparison, the non-endothelium-dependent vasodilator nitroglycerin (0.3 mg intracoronary), were infused into the left coronary arteries of 18 patients (mean age 47+/-9 years, 8 women and 10 men) with biopsy-proven myocarditis but without angiographically demonstrable coronary artery disease. Vascular responses were analyzed by quantitative coronary angiography. Three patients had an intact vasodilator response to acetylcholine concentrations of up to 10(-4) mol/L in all segments of the left coronary artery, with a mean dilatation of +9.9%+/-2%. In contrast, paradoxical constriction by acetylcholine occurred in 9 patients, who showed a mean change in coronary artery diameter of - 11%+/-3%. Six patients had no significant change in any segments in response to acetylcholine (-2.5%+/-4%). There was a significant inverse correlation between the number of T-lymphocytes in the myocardium and the response of the epicardial coronary arteries to acetylcholine (Pearson correlation coefficient -0.49, P=.03). CONCLUSIONS: It can be assumed that the process of myocarditis is associated with impairment of endothelium-dependent vasodilation in response to acetylcholine in most patients. Vasoconstriction in the presence of acetylcholine in myocarditis is likely to reflect an abnormality of endothelial function. Endothelial dysfunction of coronary arteries may explain the occurrence of myocardial ischemia in patients with myocarditis. PMID- 9736129 TI - Heterogeneity of platelet aggregation and major surface receptor expression in patients with acute myocardial infarction. AB - BACKGROUND: Platelets play an important role in the natural history of acute myocardial infarction (AMI). METHODS AND RESULTS: Platelet aggregation and receptor expression were studied in 23 patients with AMI before reperfusion therapy and compared with 10 healthy control subjects. Platelet aggregation was induced with 5 micromol/L adenosine 5'-diphosphate, 10 micromol/L ADP, 1 microg/mL collagen, 1 mg/mL thrombin, and 1.25 mg/mL ristocetin. Receptor expression was measured by flow cytometry with monoclonal antibodies to p24 (CD9), Ib (CD42b), IIb (CD41b), IIIa (CD61), IIb/IIIa (CD41b/CD61), very late antigen-2 (CD49b), P-selectin (CD62p), platelet/endothelial cell adhesion molecule-1 (CD31); and vitronectin (CD51/CD61). The percentage of platelet aggregation was higher in patients with AMI when induced by 5 micromol/L ADP (64.1+/-12.7 vs 52.0+/-6.7; P=.04), by 10 micromol/L ADP (71.7+/-13.0 vs 59.2+/ 7.2, P=.003), by thrombin (75.8+/-10.9 vs 60.5+/-6.9, P=.01), and by ristocetin (92.5+/-7.8 vs 71.3+/-7.4, P=.0001). Collagen-induced platelet aggregation did not differ between groups. Expression of P-selectin (log amplification of fluorescence intensity) (31.5+/-5.0 vs 25.1+/-2.6, P=.003) and platelet/endothelial cell adhesion molecule-1 (56.8+/-17.7 vs 44.5+/-3.7, P=.04) were significantly increased in patients with AMI. The expression of IIb (28.4+/ 2.5 vs 37.2+/-1.7, P=.0001) and Ib (103.6+/-29.9 vs 133.8+/-8.0, P=.007) were reduced in patients with AMI. CONCLUSIONS: Platelets are not necessarily systemically activated during the prereperfusion phase of AMI. For each agonist used and surface antigen measured, there was a cohort of patients with AMI within the normal or even below normal range of platelet status. PMID- 9736131 TI - Differential plasma endothelin levels in subgroups of patients with angina and angiographically normal coronary arteries. Coronary Artery Disease Research Group. AB - BACKGROUND: Raised plasma endothelin concentrations have previously been reported in patients with cardiac syndrome X, but it is not known whether these levels vary between clinically distinct subgroups in this heterogeneous condition. METHODS AND RESULTS: We compared plasma immunoreactive endothelin levels in 54 patients with angina pectoris and normal coronary angiograms and 21 healthy control subjects. The patient group was divided into 4 clinically distinct subgroups: 7 with left bundle branch block (group A); 7 with previous myocardial infarction (group B); 24 with positive exercise electrocardiography (group C); and 16 with negative exercise electrocardiography (group D). The plasma endothelin concentration was significantly higher in patients compared with control subjects (3.7 [2.9 to 4.3] vs 2.96 [2.4 to 3.4] pg/mL, respectively, median [interquartile range]; P=0.002). Endothelin concentrations were most significantly elevated in group A and group B (4.5 [3.6 to 5.2] pg/mL; P=0.005 and 4.1 [3.9 to 4.5] pg/mL; P=0.001, respectively). Plasma endothelin concentrations were also significantly elevated in group C (3.7 [2.8 to 4.1] pg/mL; P=0.02) but not in group D (3.0 [2.5 to 3.8] pg/mL; P=0.3). CONCLUSIONS: Plasma endothelin concentration is elevated in patients with angina pectoris and angiographically normal coronary arteries, particularly those with left bundle branch block or previous myocardial infarction. PMID- 9736130 TI - Pretreatment plasminogen activator inhibitor-1 (PAI-1) levels and the outcome of thrombolysis with streptokinase in patients with acute myocardial infarction. AB - BACKGROUND: The risk for reinfarctions and delays in reperfusion after streptokinase therapy may be caused by the antifibrinolytic effect of platelet derived type 1 plasminogen activator inhibitor (PAI-1). This study aims to show the relation of pretreatment PAI-1 levels of patients with acute myocardial infarction treated with streptokinase therapy and the outcome of fibrinolysis, with the emphasis on reperfusion and reinfarction. METHODS: Pretreatment PAI-1 levels of 60 patients with acute myocardial infarction, treated with streptokinase, were determined by the chromogenic method. Failure of thrombolysis with streptokinase was present when reperfusion was delayed as assessed by noninvasive reperfusion criteria, or reinfarctions developed. RESULTS: Mean pretreatment PAI-1 level of patients was 6.3+/-1.2 U/ml; span 1.2 U/ml to 34.0 U/ml. Thrombolysis with streptokinse failed significantly in patients with pretreatment PAI-1 levels >4.0 U/ml (p < 0.05), mainly because of significant occurrence of reinfarction (p < 0.05), but less to delayed reperfusion (p > 0.05). CONCLUSION: Failure of thrombolysis with streptokinase is significantly associated with pretreatment PAI-1 levels of >4.0 U/ml. PMID- 9736132 TI - Sex differences in ventricular function in patients with right bundle branch block. AB - BACKGROUND: Left ventricular function in patients with right bundle branch block is variable and depends on the population under study. This study assessed the implications of right bundle branch block for the estimation of resting left ventricular function in patients with right bundle branch and suspected coronary artery disease. METHODS AND RESULTS: Seventy-four patients with right bundle branch block, symptoms suggestive of coronary artery disease, and no electrocardiographic Q waves were compared with 649 patients with entirely normal electrocardiograms to assess the implications of right bundle branch block on resting left ventricular function. Resting ejection fraction was determined by radionuclide ventriculography. Patients with right bundle branch block were older (mean 65.0+/-10.2 years vs 53.8+/-11.1; P< .001) and had a lower mean ejection fraction (60%+/-11% vs 63%+/-9%; P< .005) compared with patients with normal electrocardiograms. There was a highly significant interaction between right bundle branch block and sex with respect to resting ejection fraction (P< .001). The mean ejection fraction for men with right bundle branch block was 57%+/-10% (17% with abnormal resting ejection fraction) compared with 62%+/-8% (7% with abnormal resting ejection fraction) for normal men. In contrast, the mean ejection fraction for women with right bundle branch block was 68%+/-9% (0% with abnormal resting ejection fraction) compared with 65%+/-9% (5% with abnormal resting ejection fraction) for normal women. CONCLUSIONS: Male patients with right bundle branch block and symptoms suggestive of coronary artery disease have a lower resting ejection fraction than mole patients with normal electrocardiograms. This difference is not seen in female patients. PMID- 9736133 TI - Heart rate changes preceding ventricular ectopy in patients with ventricular tachycardia caused by reentry, triggered activity, and automaticity. AB - OBJECTIVES: Although enhanced sympathetic tone is thought to be proarrhythmic and beta-blockade reduces the risk of sudden cardiac death in survivors of myocardial infarction, the role of the autonomic nervous system in triggering spontaneous ventricular ectopy and ventricular tachycardia (VT) has not been fully elucidated. The purpose of this study was to compare and contrast autonomic tone preceding spontaneous ventricular arrhythmias in patients with reentrant, triggered, and automatic forms of VT. BACKGROUND: The prevailing model of reentrant VT is based on a triggering beat interacting with a fixed substrate. Within this model, cyclic fluctuations in autonomic tone comprise a "third factor" that may initiate the triggering extrasystoles as well as alter the substrate, facilitating perpetuation of tachycardia. Consistent with this model, adrenergic stimulation can facilitate the induction of reentrant arrhythmias as well as arrhythmias resulting from enhanced automaticity and those caused by triggered activity resulting from cyclic adenosine monophosphate-dependent delayed afterdepolarizations. METHODS AND RESULTS: On the basis of the results at electrophysiologic study, 26 patients with coronary artery disease were identified as having reentrant VT, 11 were identified as having idiopathic VT caused by triggered activity, and 4 were identified as having idiopathic VT caused by enhanced automaticity. Each patient underwent 24-hour electrocardiographic monitoring, and the mean sinus R-R intervals immediately preceding each sinus beat as well as the 15 beats preceding sinus beats, premature ventricular contractions (VPCs), and complex ventricular ectopy (couplet/non-sustained VT) were computed. In addition, high-frequency heart rate variability was determined. Heart rate accelerated before spontaneous ventricular ectopy for all three arrhythmia mechanisms. R-R intervals preceding episodes of complex ventricular ectopy were significantly shorter than the corresponding intervals preceding single VPCs in patients with 'riggered VT [p=0.006 and 0.01, R-R(-1) and R-R(-15), respectively] and in those with reentrant VT (p=0.007 and p=0.05). There were no corresponding differences in high-frequency heart rate variability. R-R intervals preceding single VPCs were significantly shorter than the corresponding intervals preceding sinus beats in patients with automatic VT (p=0.0004 and 0.0001, respectively), which was accompanied by a small reduction in high-frequency heart rate variability (p=0.04). CONCLUSIONS: Heart rate accelerates before spontaneous ventricular ectopy in patients with VT. The acceleration is disproportionate to parasympathetic withdrawal, implicating increased endogenous sympathetic tone in the genesis of spontaneous ventricular arrhythmias caused by all three electrophysiologic mechanisms: reentry, triggered activity, and automaticity. PMID- 9736134 TI - Cardiac performance early after cardioversion from atrial fibrillation. AB - BACKGROUND: The mechanism for early improvement in cardiac function after cardioversion from atrial fibrillation is unknown. METHODS: We measured ventricular volumes and load-independent contractility during atrial fibrillation and within 24 hours after cardioversion to sinus rhythm in 15 adult patients (10 men, 5 women; mean age 63+/-4 years, range 31 to 81 years). Duration of atrial fibrillation ranged from <1 day to 6 months. RESULTS: After cardioversion, left ventricular ejection fraction increased from 51%+/-4% to 61%+/-4% (P=.001, 95% confidence intervals for the difference, 7% to 15%), stroke volume increased from 57+/-4 mL to 76+/-6 mL (P < .001, 95% confidence intervals 8 to 32 mL), and mean cycle length increased from 0.77+/-.04 seconds in atrial fibrillation to 1.02+/ .04 seconds in sinus rhythm (P=.002, 95% confidence intervals, 0.1 to 0.4 seconds). Cardiac contractility, as expressed by the slope and the intercept of the relation between rate-corrected circumferential velocity of fiber shortening and end-systolic wall stress (Vcfc/ESWS) remained unaltered in 13 of 15 patients, suggesting that intrinsic inotropic state was unchanged immediately after return of normal sinus rhythm. Finally, a significant correlation was observed between improvement in stroke volume and peak A-wave velocity (r=0.79, P=.035). CONCLUSION: Both left ventricular stroke volume and ejection fraction increase immediately after cardioversion, whereas intrinsic cardiac contractility is largely unchanged. These data suggest that the mechanism of this increase is enhanced left ventricular diastolic filling due mostly to increased cycle length and return of left atrial mechanical function. PMID- 9736135 TI - Autonomic functions in restrictive cardiomyopathy and constrictive pericarditis: a comparison. AB - BACKGROUND: This study was undertaken to analyze autonomic functions in restrictive cardiomyopathies. Restrictive cardiomyopathies have clinical and hemodynamic similarity with chronic constrictive pericarditis. Autonomic dysfunction has been described in the latter. METHODS AND RESULTS: Autonomic function analysis has not been reported in restrictive cardiomyopathy. Six consecutive patients with restrictive cardiomyopathy were included in this study (5 men, 1 woman, mean age 35+/-5.4 years). The tests performed were designed to test the sympathetic efferent pathway, that is, by cold hand immersion and loud noise tests, parasympathetic efferent pathway by Valsalva ratio and expiration/inspiration ratio and the baroreceptor function by testing their sensitivity slope. The results were compared with 20 patients with chronic constrictive pericarditis and with 10 healthy age- and sex-matched control subjects previously studied. The rise of systolic blood pressure after cold hand immersion and sudden loud noise was not significantly different compared with control subjects. The expiration/inspiration ratio was 1.1+/-0.01 compared with 1.57+/-0.1 in the control group (p < 0.01). The Valsalva ratio was significantly lower (1.1+/-0.04) compared with control subjects (1.83+/-0.1, p < 0.01). The baroreceptor sensitivity was not reduced compared with that in control subjects. In comparison to constrictive pericarditis, sympathetic efferent pathway is preserved in restrictive cardiomyopathy (p < 0.0001). The parasympathetic efferent pathway is borderline abnormal in restrictive cardiomyopathy but not significantly as compared with constrictive pericarditis (p=not significant). The baroreceptor sensitivity slope is normal in patients with restrictive cardiomyopathy as compared with significant depression seen in constrictive pericarditis (p < 0.05). Autonomic functions are better preserved in patients with restrictive cardiomyopathies compared with chronic constrictive pericarditis. CONCLUSIONS: Autonomic dysfunction is localized to parasympathetic efferent pathway. This is in comparison to constrictive pericarditis, in which severe autonomic dysfunction is a universal feature and includes all segments of autonomic nervous system. PMID- 9736136 TI - Does the 6-minute walk test predict the prognosis in patients with NYHA class II or III chronic heart failure? AB - BACKGROUND: We prospectively evaluated the potential of the 6-minute walk test compared with peak VO2 in predicting outcome of patients with New York Heart Association (NYHA) class II or III heart failure. METHODS AND RESULTS: Patients with a history of heart failure caused by systolic dysfunction were included. The combined final outcome (death or hospitalization for heart failure) was used as the judgment criterion. One hundred twenty-one patients (age 59+/-11 years; left ventricular ejection fraction 29.6%+/-13%) were included and followed for 1.53+/ 0.98 years. Patients were separated into two groups according to outcome: group 1 (G1, 74 patients), without events, and group 2 (G2, 47 patients), who reached the combined end point. Peak VO2 was clearly different between G1 and G2 (18.5+/-4 vs. 13.9+/-4 ml/kg/min, p=0.0001) but not the distance walked (448+/-92 vs 410+/ 126 m; p=0.084, not significant). Survival analysis showed that unlike peak VO2, the distance covered was barely distinguishable between the groups (p < 0.08). However, receiver operating characteristic curves revealed that the best performances for the 6-minute walk test were obtained for subjects walking < or =300 m. These patients had a worse prognosis than those walking farther (p=0.013). In this subset of patients, there was a significant correlation between distance covered and peak VO2 (r=0.65, p=0.011). Thus it appears that the more severely affected patients have a daily activity level relatively close to their maximal exercise capacity. Nevertheless, the 300 m threshold suggested by this study needs to be validated in an independent population. CONCLUSIONS: A distance walked in 6 minutes < or =300 m can predict outcome. Moreover, in these cases there is a significant correlation between the 6-minute walk test and peak VO2 demonstrating the potential of this simple procedure as a first-line screening test for this subset of patients. PMID- 9736137 TI - Importance of intrinsic calf vasodilator capacity in determining distribution of skeletal muscle perfusion during supine bicycle exercise in patients with left ventricular dysfunction. AB - BACKGROUND: Distribution of skeletal muscle perfusion during exercise is an important factor in determining exercise capacity and is markedly impaired in patients with cardiac disease. This study examined the importance of intrinsic calf vasodilator capacity in determining distribution of skeletal muscle perfusion during supine bicycle exercise in patients with left ventricular dysfunction. METHODS: We studied 19 patients with left ventricular dysfunction (left ventricular ejection fraction <45%) after myocardial infarction. All the patients underwent cardiopulmonary exercise testing with measurements of central hemodynamics, leg blood flow (LBF), and the percentage of cardiac output distributed to both legs (%LBF). Calf reactive hyperemic flow (RH) was measured by venous occlusive plethysmography at supine rest. RESULTS: LBF at peak exercise was closely related to peak cardiac output and RH. Furthermore, %LBF at peak exercise had modest correlation with peak cardiac output and good correlation with RH. Although peak cardiac output and RH were independent determinants of LBF at peak exercise by multiple regression analysis, RH had higher correlation with %LBF at peak exercise than peak cardiac output. Despite marked changes in other hemodynamic variables, nonleg blood flow during exercise was constantly maintained at a level identical to resting value. CONCLUSIONS: Calf vasodilator capacity, which was the major determinant of distribution of skeletal muscle perfusion during exercise, may have contributed to maintaining perfusion of important nonexercising regions during exercise in patients with left ventricular dysfunction. PMID- 9736138 TI - Impaired exercise capacity late after cardiac transplantation: influence of chronotropic incompetence, hypertension, and calcium channel blockers. AB - BACKGROUND AND METHODS: Patients undergoing orthotopic cardiac transplantation manifest reduced exercise capacity during the first postoperative year, which is related primarily to chronotropic incompetence of the denervated heart. To determine whether exercise capacity improves during the long term after transplantation, we prospectively studied 45 patients from 1 month to 6 years after cardiac transplantation by use of maximal treadmill exercise testing for measurement of exercise duration, peak heart rate, and peak VO2. All had normal left ventricular ejection fractions. Patients were categorized according to length of time since transplant and compared to 14 untrained normal subjects. RESULTS: Peak exercise heart rate and exercise duration were progressively higher as time after transplantation increased. However, patients who had undergone transplantation more than 2 years earlier continued to manifest a significant reduction in peak exercise heart rate (157+/-3 beats/min vs 178+/-14 beats/min) and reduced exercise duration (8.6+/-0.5 minutes vs 13.2+/-2.0 minutes) compared with controls. In contrast, peak VO2 was similar at all times after transplant and remained markedly reduced in patients who underwent transplantation more than 2 years earlier as compared with controls (22.1+/-0.7 mL/kg/min vs 42.1+/-9.1 mL/kg/min). The potential effects of 14 clinical variables on exercise performance were evaluated by regression modeling. Patients with poorly controlled hypertension had a shorter median exercise duration (7.4 minutes vs 9.7 minutes) and a lower median peak VO2 (20.3 mL/kg/min vs 23.2 mL/kg/min) compared with patients with normal or well-controlled blood pressure. Patients treated with calcium channel blockers for hypertension had greater chronotropic incompetence during exercise (peak heart rate 139 beats/min vs 158 beats/min). There was no relation between exercise capacity and recipient age, donor age, recipient sex, donor ischemic time, pretransplant diagnosis, length of peritransplant hospitalization, percentage of ideal body weight, left ventricular ejection fraction, frequency or severity of allograft rejection, or long-term use of oral prednisone therapy. CONCLUSIONS: Exercise capacity, as measured by treadmill exercise time and peak heart rate, improves in the first 2 years after transplantation, but does not reach normal values in patients up to 6 years after transplant. Peak VO2 remains significantly reduced at all times after transplantation despite the presence of normal resting left ventricular systolic function. PMID- 9736139 TI - Evidence of left ventricular dysfunction in children with merosin-deficient congenital muscular dystrophy. AB - BACKGROUND: Deficiency of the sarcolemmal protein dystrophin has been linked to dilated cardiomyopathy. Some children with congenital muscular dystrophy have a deficiency of the laminin alpha2 chain of merosin, an extracellular matrix protein linked to dystrophin through a group of glycoproteins. It has been shown that deficiency in one of these glycoproteins is responsible for muscular dystrophy and dilated cardiomyopathy. Children with laminin alpha2 deficiency may be at risk for development of cardiomyopathy. METHODS AND RESULTS: We studied the cardiac function of a cohort of 16 children with congenital muscular dystrophy by using 2-dimensional echocardiography. The expression of the laminin alpha2 of merosin in the patients was determined on a skin or muscle biopsy. Two of 6 merosin-deficient children had an ejection fraction <40%. The average ejection fraction of the merosin-deficient children was 43%+/-11%, which was significantly lower than the merosin-positive children (53%+/-5%, P=.03). CONCLUSIONS: This study suggests that a deficiency of laminin alpha2 can give rise to dilated cardiomyopathy, supporting the idea that defects of dystrophin, or of associated proteins, can cause dilated cardiomyopathy in addition to muscular dystrophy. PMID- 9736140 TI - How does warfarin affect the activated coagulation time? AB - BACKGROUND: The activated coagulation time (ACT) is a rapid measurement of a patient's level of heparin anticoagulation during cardiac catheterization. Patients receiving warfarin therapy occasionally are seen at the catheterization laboratory for emergent procedures. The effects of warfarin on ACT activity have not been previously described. We compared the ACT and the international normalized ratios (INR) in 77 patients receiving warfarin and 57 patients who were not receiving any anticoagulation (controls). RESULTS: Both the mean ACT (131+/-17.0 seconds) and INR (2.5+/-0.90 seconds) of the anticoagulated patients differed from the controls (ACT=115+/-14.5 seconds, INR=1.0+/-0.10 seconds; P< 0.05). The ACT increased linearly with INR in the warfarin group (r=0.70, P< .001). There was no relation between ACT and INR in the control group. CONCLUSION: Patients receiving warfarin therapy will have a linear increase in ACT develop similar to patients receiving heparin therapy. PMID- 9736141 TI - Is low magnesium concentration a risk factor for coronary heart disease? The Atherosclerosis Risk in Communities (ARIC) Study. AB - BACKGROUND: Hypomagnesemia has been hypothesized to play a role in coronary heart disease (CHD), but few prospective epidemiologic studies have been conducted. METHODS AND RESULTS: We examined the relation of serum and dietary magnesium with CHD incidence in a sample of middle-aged adults (n=13,922 free of baseline CHD) from 4 US communities. Over 4 to 7 years of follow-up, 223 men and 96 women had CHD develop. After adjustment for sociodemographic characteristics, waist/hip ratio, smoking, alcohol consumption, sports participation, use of diuretics, fibrinogen, total and high-density lipoprotein cholesterol levels, triglyceride levels, and hormone replacement therapy, the relative risk of CHD across quartiles of serum magnesium was 1.00, 0.92, 0.48, and 0.44 (P for trend=0.009) among women and 1.00, 1.32, 0.95, and 0.73 (P for trend=0.07) among men. The adjusted relative risk of CHD for the highest versus the lowest quartile of dietary magnesium was 0.69 in men (95% confidence interval 0.45 to 1.05) and 1.32 in women (0.68 to 2.55). CONCLUSIONS: These findings suggest that low magnesium concentration may contribute to the pathogenesis of coronary atherosclerosis or acute thrombosis. PMID- 9736143 TI - QT dispersion may be a useful adjunct for detection of myocardial infarction in the chest pain center. AB - BACKGROUND: QT dispersion has been proposed as a noninvasive measurement of the degree of inhomogeneity in myocardial repolarization. Increased QT dispersion has been reported after myocardial infarction. We hypothesized that increased QT dispersion may be a useful adjunct for risk stratification in patients being evaluated in a chest pain center. METHODS AND RESULTS: Patients were admitted to the chest pain center for evaluation of chest pain. Exclusion criteria included (1) systolic blood pressure <90 mm Hg, (2) ischemia or infarction on the initial electrocardiograph (ECG), (3) elevated creatine kinase or MB fraction, and (4) chest pain associated with cocaine use. Serial creatine kinase and MB levels and ECGs were obtained at 0, 6, and 9 hours. Patients were monitored for (1) creatine kinase and MB rise, (2) ECG changes for infarction, (3) ST-segment changes, and (4) rest angina. A negative evaluation at the chest pain center led to an exercise stress test. Patients with a positive exercise stress test were admitted for further evaluation and patients with a negative exercise stress test result were discharged home. Patients were divided into 3 groups. Group 1 consisted of patients who were found to have an acute myocardial infarction (AMI), group 2 consisted of patients with prior history of coronary artery disease but no evidence of AMI, and group 3 consisted of patients without prior coronary artery disease or AMI. QT dispersion was measured on the initial ECG in all patients. A total of 586 patients were evaluated. Group 1 consisted of 13 patients with mean QT dispersion of 44.6+/-18.5 ms, group 2 consisted of 267 patients with a mean QT dispersion of 10.0+/-13.8 ms, and group 3 consisted of 303 patients with a mean QT dispersion of 10.5+/-10.0 ms. Analysis of variance showed a significantly higher QT dispersion in patients who had AMI compared with other patients with chest pain (P< .001). CONCLUSIONS: QT dispersion can be a useful diagnostic adjunct for detection of AMI in patients with chest pain with a normal ECG and normal cardiac enzymes. PMID- 9736142 TI - Depression and social support in elderly patients with cardiac disease. AB - BACKGROUND: Depression is common among patients with cardiac disease. A number of psychosocial factors may affect the relationship between physical health and depression. There is evidence from the psychiatric literature suggesting that negative life events and social support are important factors in the development and outcome of depression. It is unknown if these factors are important in the context of depression in medically ill patients. Thus it is important to examine the relationship among social support, negative life events, and the presence of depression in elderly patients with cardiac disease. METHODS: Patients with coronary artery disease were assessed with the Duke Depression Evaluation Schedule for the Elderly. This includes the mood and anxiety disorder section of the Diagnostic Interview Schedule modified for Diagnostic and Statistical Manual of Mental Disorders diagnoses, life events, and multidimensional assessment of social support. Two hypotheses were tested: (1) the number of concurrent negative life events will be higher in patients with coronary artery disease with major depression than those without depression, and (2) social support will be less in patients with major depression than in those without. RESULTS: Presence of major depression was associated with increased negative life events and lowered subjective social support after accounting for age, sex, and race. CONCLUSIONS: The finding that subjective social support and negative life events are related to major depression suggests that even in the context of medical illness, social factors are still important in the development of major depression. PMID- 9736144 TI - Exaggerated blood pressure response at exercise in normotensive subjects: demographic and stress performance characteristics. AB - BACKGROUND: Exercise testing is an important diagnostic and prognostic procedure in the assessment of patients with hypertension. An exaggerated blood pressure response to exercise among normotensive subjects was found to be one of the best predictors of future hypertension. The demographic characteristics of patients with an exaggerated blood pressure response during exercise have not been adequately described. METHODS AND RESULTS: The demographic and stress performance characteristics of 2 groups of normotensive patients referred for exercise testing, one composed of patients with an exaggerated blood pressure response (group I, n=146) and a group of patients with a normal blood pressure response (group II, n=439) were prospectively compared. Patients in group I were older than those in group II (54+/-12 vs 51+/-13 years, P < .05). More men than women were found in both groups, yet significantly more in group I than in group II (83% vs 69% P < .001). Significantly more among the patients in group I had a higher level of education and were of Western origin than those in group II (P < .01). The resting systolic and diastolic blood pressures were higher in group I than in group II (131+/-18 vs 119+/-14 mm Hg, P< .001, and 81+/-8 vs 76+/-7 mm Hg, P < .001, respectively). The patients in group I achieved a higher percentage of the maximal predicted heart rate (88+/-7 vs 85+/-9 beats/min, P < .01). No significant differences were found between the groups in the duration of stress test and effort ischemia. CONCLUSIONS: Patients with a hypertensive blood pressure response during stress testing have specific demographic and exercise characteristics. PMID- 9736145 TI - Progressive deterioration of coronary flow reserve after heart transplantation. AB - BACKGROUND: The purpose of this study was to determine coronary flow reserve in cardiac allograft recipients early (0 to 3 years) and late (3 to 7 years) after heart transplantation. METHODS AND RESULTS: With the use of a Doppler tipped guide wire, coronary flow reserve (ratio of hyperemic to baseline coronary flow velocity) was measured in 82 patients before and after intracoronary adenosine. Forty-five patients were early (< or =3 years) after transplantation, 24 were late, and 13 were control patients with single-vessel coronary artery disease. Coronary flow reserve in the early transplantation patients was similar to that in the control group (2.9+/-0.2 vs 3.0 +/-0.6, p=not significant) but was reduced in the late transplantation group (2.2+/-0.5 vs 3.0+/-0.6, p < 0.001). There were differences in coronary flow reserve between the early and late transplantation patient groups (3.0+/-0.6 vs 2.2+/-0.5, p < 0.001 ) despite equally elevated mean arterial pressure, mean heart rate, mean pulmonary capillary wedge pressure, and mean left ventricular mass in the two groups. Coronary flow reserve in patients with angiographic allograft arteriopathy (n=19) was reduced when compared with coronary flow reserve of patients with normal vessels (n=50) (1.9+/-0.3 vs 3.1+/ 0.6, p < 0.001). CONCLUSIONS: There is progressive deterioration of coronary flow reserve over time after transplantation. Dysfunction of the coronary microcirculation rather than determinants of myocardial oxygen consumption contributes to this reduction. PMID- 9736146 TI - Emotional distress before coronary bypass grafting limits the benefits of surgery. AB - BACKGROUND: The inclusion of large, heterogeneous groups of patients for coronary bypass grafting (CABG) surgery has resulted in a more mixed treatment outcome. Thus it becomes important to identify patients who are less likely to benefit from surgery or who may require additional support to improve treatment outcome. The aim of the present study was to examine whether psychological status measured before CABG can contribute to prediction of short- and long-term outcomes of the surgery. METHODS AND RESULTS: One hundred seventy-one consecutive patients from two large university hospitals in Stockholm completed a psychosocial questionnaire before being scheduled for surgery. One year after CABG, patients again completed the questionnaire. Follow-up of medical charts was conducted during the first 3 years after surgery. All major cardiac events (cardiac death, definite myocardial infarction, revascularization, and unstable angina verified by angiography or myocardial scintigraphy) were recorded. Although the overall effect of surgery was excellent in the majority of cases, the patients exhibiting a high degree of distress (anxiety, depression, and tiredness) before surgery assessed their status as being much worse both before the operation and at the 1 year follow-up. Equally important was the fact that patients considered distressed before surgery had significantly higher rates of cardiac events (16%) in the 3-year follow-up period compared with nondistressed patients (5%) (chi square=5.11, degrees of freedom=1, p < 0.02). CONCLUSIONS: Systematic evaluation and treatment of emotional distress in the candidates for coronary revascularization may be expected to result in more optimal subjective results and a reduction in the number of serious cardiac events after surgery. PMID- 9736147 TI - Incidence and predictors of restenosis after successful primary coronary angioplasty for acute myocardial infarction: the importance of age and procedural result. AB - BACKGROUND: Previous studies have suggested that restenosis and reocclusion occur frequently in patients with acute coronary syndromes. This study was undertaken to assess the incidence and predictors of restenosis in a cohort of patients who underwent successful primary coronary angioplasty for acute myocardial infarction. METHODS: Three hundred twelve patients who underwent successful primary angioplasty of a native coronary vessel were candidates for follow-up coronary angiography. This was performed in 284 patients (92%) at the 3- or 6 month follow-up. Quantitative coronary angiography was performed with the CMS system. Multivariate analysis was performed to determine independent predictors of restenosis. RESULTS: Restenosis, defined as a diameter stenosis of >50%, occurred in 27% of patients at 3 months and in 37% of patients at 6-month follow up. Reocclusion occurred in 4% and 6%, respectively. Reference diameter (vessel size) was related to restenosis. Age and lumen diameter immediately after angioplasty were independent predictors of restenosis. Young patients (<50 years) and patients with a minimal luminal diameter of more than 2.5 mm had restenosis rates of <25%. The radionuclide ejection fraction was 46% in patients with restenosis compared with 47% in patients without restenosis. CONCLUSIONS: The incidence of restenosis after successful primary coronary angioplasty for acute myocardial infarction is comparable to the reported incidence after elective coronary angioplasty for stable angina. Restenosis is related to age and the lumen diameter after angioplasty and does not affect left ventricular function in this population. PMID- 9736148 TI - Mitral regurgitation reduces the risk of stroke in patients with nonrheumatic atrial fibrillation. AB - BACKGROUND: Significant mitral regurgitation (MR) is protective against left atrial (LA) spontaneous echo contrast formation that is associated with an increased thromboembolic risk. However, the effects of MR on the risk of stroke in patients with nonrheumatic atrial fibrillation (AF) have been unknown. We studied whether or not MR was associated with a decreased risk of stroke in patients with nonrheumatic AF. METHODS: We performed an observational analysis of retrospectively collected data on 290 patients with nonrheumatic AF. Left atrial diameter (LAD) and the degree of MR were estimated by transthoracic echocardiography. Risk factors for stroke were assessed by univariate and multivariate analyses. The mean follow-up was 7.4 years. RESULTS: Among these patients, 68 had a stroke during the follow-up (rate of stroke per year of follow up 3.2%). In 95 patients with LAD of > or =48 mm, the incidence of stroke (9%) in the severe MR group (moderate or severe, n=43) was significantly lower than that (25%) of the mild MR group (none, trivial, or mild; n=52) (chi-square=3.95, p=0.047). The relative risk of stroke for increase in MR from mild to severe groups, for every 10 mm increment in LA size, for sex, and for every increase of 10 years of age was 0.45 (95% CI, 0.20 to 0.97), 1.06 (95% CI, 0.75 to 1.49), 0.98 (95% CI, 0.55 to 1.72), and 1.33 (95% CI, 1.04 to 1.71), respectively. CONCLUSIONS: In patients with nonrheumatic AF, age was an independent predictor of an increased risk of stroke, and MR may be protective against stroke, especially in those patients with LA enlargement. PMID- 9736149 TI - Correlation between exercise-induced ischemic ST-segment depression and myocardial blood flow quantified by positron emission tomography. AB - BACKGROUND: Ischemic ST-segment depression (horizontal or downsloping) is the most common manifestation of exercise-induced myocardial ischemia. The mechanisms responsible for these types of ST-segment depression are largely unknown. We investigated the relation of these 2 types of exercise-induced ST-segment depression to changes in regional myocardial blood flow (RMBF) by using exercise positron emission tomography (PET). METHODS AND RESULTS: The RMBF was measured with nitrogen-13 ammonia PET both at rest and during low-level supine bicycle exercise in 27 patients with angiographically proven coronary artery disease and in 6 healthy volunteers. ST-segment depression was measured from the isoelectric PR segment 80 ms after the J point. Exercise-induced horizontal ST-segment depression > or =0.1 mV was observed in 9 patients and downsloping depression > or =0.1 mV was observed in 18 patients. Multivessel disease was more frequent and areas of exercise-induced ischemia were larger in patients with downsloping depression than in patients with horizontal depression (P < .02, P < .05). In patients with horizontal ST-segment depression, RMBF in ischemic areas and in surrounding areas increased by a similar amount (31%+/-29% and 32%+/-16%) with exercise. In patients with downsloping ST-segment depression, RMBF was unchanged or decreased in ischemic areas (10%+/-24%) but increased in surrounding areas (46%+/-27%) with exercise. In healthy volunteers, RMBF increased in all areas (56%+/-30%) with exercise. CONCLUSIONS: Compared with horizontal changes in ST segment morphology, downsloping changes may better indicate severe ischemia and greater differences in the increase of blood flow with exercise in the ischemic myocardium and in the surrounding areas. PMID- 9736150 TI - Can computerization of the exercise test replace the cardiologist? AB - BACKGROUND: The type of practitioners who use the standard exercise test is changing. Once a tool of the cardiologist, the standard exercise test is now being performed by internists and other noncardiologists. Because this change could be facilitated by computerization similar to the computerized interpretation programs available for the resting electrocardiograph (ECG), we performed this analysis. A secondary aim was to demonstrate the effects of medication status and resting ECG abnormalities on test diagnostic characteristics because these factors affect utility of the exercise test by the generalist. METHODS AND RESULTS: A retrospective analysis was performed of consecutive patients referred at 2 university-affiliated Veteran's Affairs Medical Centers and a Hungarian Hospital for evaluation of chest pain and possible ischemic heart disease. There were 1384 consecutive male patients without a prior myocardial infarction with complete data who had exercise tests and coronary angiography between 1987 and 1997. Measurements included clinical, exercise test data, and visual interpretation of the ECG recordings as well as more than 100 computed measurements from the digitized ECG recordings and compilation of angiographic data from clinical reports. The computer measurements had similar diagnostic power compared with visual interpretation. Computerized measurements from maximal exercise or recovery were equivalent or superior to all other measurements. Prediction equations applied by computer were superior to single ECG measurements. Beta-blockers had no effect on test characteristics, whereas resting ST depression was associated with decreased specificity and increased sensitivity. CONCLUSIONS: Computerized exercise ST measurements are comparable to visual ST measurements by a cardiologist; computerized scores that included clinical and exercise test results exhibited the greatest diagnostic power. Applying scores with a computer allows the practicing physician to improve the diagnostic characteristics of the standard exercise test. This approach is successful even when there is resting ST depression, thus lessening the need for more expensive nuclear or imaging studies. PMID- 9736151 TI - Managed care for congestive heart failure: influence of payer status on process of care, resource utilization, and short-term outcomes. AB - BACKGROUND: Although health maintenance organizations (HMO) are insuring an increasing number of Americans, there are concerns that cost-reduction strategies may limit access to medical care or jeopardize its quality. This study was conducted to examine the influence of insurance payer status on the process of care and resource utilization among patients hospitalized for congestive heart failure (CHF). METHODS AND RESULTS: Administrative information on all 1995 New York State hospital discharges assigned ICD-9-CM codes indicative of CHF in the principal diagnosis position were obtained from the Statewide Planning and Research Cooperative System database. The following were compared among patients with HMO, indemnity, Medicaid fee-for-service, and Medicare fee-for-service insurance coverage: demographics, comorbid illness, process of care, length of stay, hospital charges, mortality rate, and CHF readmission rate. A total of 43,157 patients were identified (HMO, 1322; indemnity, 4350; Medicaid, 3878; Medicare, 33 607). Noninvasive procedures were used with similar frequency, whereas greater use of invasive techniques was observed among HMO and indemnity patients. After adjustment for patient characteristics and hospital type and location, HMO care was associated with shorter length of stay and lower hospital charges, the latter partially explained by fewer hospital days. Medicaid patients had the longest length of stay, greatest hospital charges, and highest CHF readmission rate. The adjusted risk of death during the index hospitalization did not vary among insurance groups. CONCLUSIONS: Though insuring only a small proportion of New Yorkers hospitalized for CHF, managed care plans provide similar access to clinical services while generating fewer charges. Whether these observed differences in short-term outcomes derive from patient mix or quality of care is uncertain and deserves wider prospective study. PMID- 9736152 TI - Angiotensin-converting enzyme gene polymorphisms. PMID- 9736153 TI - C-reactive protein in postinfarction heart rupture. PMID- 9736154 TI - Lords a-leaping: the Soulsby report on antimicrobial drug resistance. PMID- 9736155 TI - Resistance to antimicrobial agents: a personal view. AB - Problems of antimicrobial drug resistance are presently serious, but not yet desperate. The principal areas of concern are two-fold: multiresistant opportunist bacteria that affect vulnerable patients in high dependency areas of hospitals (the most pressing problem for developed countries); and multidrug resistance among classic pathogens like Mycobacterium tuberculosis, Salmonella typhi, Shigella spp., Neisseria gonorrhoeae and Plasmodium falciparum (mainly, although not exclusively, a problem for developing countries). The first type can be contained to a large extent by good infection control practices and careful prescribing based on agreed policies of antimicrobial drug use. The input of infection control nurses and laboratory-based clinical microbiologists is crucial and these services deserve full support. The second type additionally requires coordinated action to regulate more effectively the manufacture, availability, promotion and use of antimicrobial drugs. In this case the input of governments, international agencies and pharmaceutical companies is essential. Prescription only status for antimicrobial drugs used in man and animals should be the norm. The number of drugs available for the treatment of viral, fungal and parasitic infections is comparatively small and much less is known about resistance. More research in these areas would be welcome. Teaching good prescribing habits to medical students is presently haphazard and needs to be formalised. Surveillance needs to be improved. The second half of the 20th century has been a golden age of antibiotics, but the outlook is uncertain. If antimicrobial chemotherapy is to have a secure future, prescribers must learn to use these powerful tools with greater discretion and their use worldwide must be regulated effectively. PMID- 9736156 TI - Analysis of TbpA and TbpB functionality in defective mutants of Neisseria meningitidis. AB - Iron uptake analysis suggested that the Neisseria meningitidis transferrin (Tf) binding proteins, TbpA and TbpB, form only one type of receptor complex. Mutants defective in the synthesis of either TbpA or TbpB, but not defective in both proteins, can bind Tf, suggesting that both proteins are surface exposed and function in Tf binding. Also, iron uptake from Tf into the meningococci did not require the presence of both Tbps. The TbpB-defective mutant incorporated c. 37% of the iron taken up by the wild-type strain, but this was insufficient for bacterial growth. The TbpA-defective mutant incorporated c. 50% of the iron taken up by the wild-type strain and was able to grow with Tf as the only iron source. Mouse antibodies specific for TbpA were able to block c. 70% of the iron uptake from Tf in the wild-type strain, whereas they blocked only 22% of iron uptake in the TbpB-defective mutant and did not block uptake in the TbpA-defective strain. These results emphasise that TbpA should be considered in future vaccine trials in which iron-restricted proteins are to be included in the vaccine formulation. PMID- 9736157 TI - Carbon source utilisation by Bordetella bronchiseptica. AB - Bordetella bronchiseptica isolates utilised tricarboxylic acid cycle intermediates -- succinate, citrate, alpha-ketoglutarate, fumarate, lactate and oxalo-acetate; the organic acids pyruvate, acetate and lactate; and the amino acids proline, glutamate, glutamine and tyrosine -- as sole sources of carbon and energy. The inability of B. bronchiseptica isolates, representing the three phase types and from different animal hosts, to utilise carbohydrates and sugar alcohols as sole carbon and energy sources was confirmed and extended. The influence of the carbon substrate on doubling time, piliation, flagellation, motility, capsule production and adherence to mammalian cells was also measured. PMID- 9736158 TI - Inhibition of enhanced toxin production by Clostridium difficile in biotin limited conditions. AB - Production of toxins A and B by Clostridium difficile is enhanced in a defined medium with biotin-limited conditions. In the present study compounds inhibitory to enhanced toxin production by a C. difficile strain were examined. Increases in biotin concentration from 0.05 nM to 50 nM accelerated growth and inhibited enhanced toxin production. Asparagine, glutamic acid and glutamine (10 mM) showed an effect on growth and toxin production similar to that of biotin. Lysine (10 mM) suppressed growth and inhibited toxin production. Addition of these toxin inhibitory compounds within an incubation period of 2 days inhibited the enhanced toxin production, but later addition showed only slight inhibition of toxin production. Amino acids contained in the defined medium under the biotin-limited conditions were actively utilised in the presence of the three toxin-inhibitory amino acids, but in the presence of lysine, amino-acid utilisation was suppressed. Different mechanisms of action of these toxin-inhibitory molecules, which may be divided into excess biotin, asparagine-glutamic acid-glutamine group, and lysine, are discussed. PMID- 9736159 TI - Differential efficacy of passive immunisation against infection by Lyme disease spirochaetes transmitted by partially fed vector ticks. AB - The efficacy of passive immunisation against tick-transmitted Lyme disease spirochaetal infection was determined in relation to the duration of previous feeding of infected vector ticks. Thus, mice challenged with spirochaete-infected unfed or partially fed nymphal ticks were passively immunised with monoclonal and polyclonal antibodies against the Lyme disease spirochaete (Borrelia burgdorferi) at various intervals after tick attachment. Spirochaetal infection in challenged mice and engorged ticks was verified by xenodiagnosis and indirect immunofluorescent antibody assay, respectively. Although tick-transmitted spirochaetal infection could be aborted by anti-OspA antibodies and hyperimmune antiserum, nearly all immunised mice challenged with infected ticks that had previous 36-h attachment became infected. More than 72% of the nymphal ticks used in this challenge retained their B. burgdorferi infection after engorgement on mice immunised with anti-spirochaete antibodies, and their subsequent infectivity to mice remained effective. It is concluded that a higher efficiency of transmission by partially fed infected nymphs and a lower efficacy of passive immunisation against infection result from an effect of previous feeding of infected ticks that activates antigenic change and enables the spirochaetes to circumvent OspA-based humoral immunity. PMID- 9736160 TI - Enteropathogenicity markers in Escherichia coli isolated from infants with acute diarrhoea and healthy controls in Rio de Janeiro, Brazil. AB - Faeces from urban children < 2 years old with acute diarrhoeal illness and from non-diarrhoeal infants (controls) were examined for Escherichia coli and other enteropathogens. A total of 990 E. coli isolates from 100 patients and 50 controls was tested for enteropathogenic E. coli (EPEC) serotype (O:H), adherence to HEp-2 cells after incubation for 3 and 6 h, fluorescent actin staining (FAS), DNA hybridisation with EAF, eaeA, STh, STp and EAggEC probes and production of heat-labile enterotoxin (LT) and verocytotoxin (VT) with Y1 and Vero cells. EPEC were the most prevalent enteropathogens in patients (32.7%; and 14% in controls). Enteroinvasive E. coli (EIEC) and Vero cytotoxin-producing E. coli (VTEC) were not detected. The rate of isolation of enterotoxigenic E. coli (ETEC) was identical in both groups. Among the EPEC isolates the prevalent serotypes were O111:H2, O55:NM and O119:H6. Localised adherence (LA) was found significantly more frequently in isolates from patients (19.6%) than controls (2.1%). All LA positive EPEC isolates were FAS+ and eaeA+, but only 75.2% of them hybridised with the EAF probe. Diffusely adhering E. coli (DAEC) and enteroaggregative E. coli (EAggEC) were found with equal frequency in patients and controls. Twenty seven E. coli isolates were negative for EAF but positive for eaeA and FAS and produced LA in 6-h adherence tests. These EAF-/eaeA+ strains were the only putative enteropathogen identified in seven patients and were not found in controls. The ability of these strains to elicit ultrastructural cell alterations and cell-signalling events was evaluated in Caco-2 cells (human colon carcinoma cell line) by the gentamicin invasion assay and by transmission electron microscopy. The numbers of intracellular bacteria in cell invasion tests varied from 0.4% to 1.6% of the cell-associated bacteria after a 6-h incubation period. Tyrosine phosphorylation of host cell proteins was assessed in HEp-2 cells by immunofluorescence microscopy and all strains gave positive results. EAF-/eaeA+ E. coli strains express most of the virulence properties found among true EPEC strains and can be a relevant cause of infant diarrhoea in developing countries. PMID- 9736161 TI - Distribution of insertion sequence IS200 among different clonal lines of the related Salmonella serotypes Livingstone and Eimsbuettel. AB - The copy number and genetic location of IS200 have provided evidence of strain relatedness in many serotypes of Salmonella. In this study, 100 isolates of the related serotypes Livingstone (6,7:d:l,w) and Eimsbuettel (6,7,14:d:l,w), representing 10 ribotype/biotype (RT/BT) groups isolated from human and non-human sources in seven countries over a 26-year period, were examined for their IS200 profiles. The distribution of IS200 in strains of these serotypes was limited, being present in all 53 isolates of ribotype 1 (RT1) and its variant type RT6, in one of five isolates of RT5 but in none of 42 isolates of RTs 2, 3 or 4. Although the seven IS200 profiles identified in RT1 isolates were of little value for further discrimination within different biotype groups, they were extremely valuable for confirming serotype: isolates of RT1/BT8/IS200 profile A (or its variants) and those of RT1/BT3/IS200 profile B (or its variants) were almost invariably associated with serotypes Livingstone and Eimsbuettel, respectively. PMID- 9736162 TI - Experimental campylobacter infection and diarrhoea in immunodeficient mice. AB - The responses of previously untested immunodeficient mouse strains to campylobacter infection are described. Three strains of adult immunodeficient mice (SCID-Beige, C.B-17-SCID-Beige and RAG-2) were inoculated intragastrically with Campylobacter jejuni NCTC 11168. All mice became heavily colonised, but none developed clinical signs of disease. Immunocompetent BALB/c mice inoculated similarly had much lower colonisation levels. The co-administration of iron dextran had no effect on colonisation levels nor the development of clinical signs of disease. In contrast, C.B-17-SCID-Beige mice, when inoculated with one of a series of 10 clinical isolates of C. jejuni, were more heavily colonised for extended periods (up to 5 months) and approximately 10-20% of the mice became ill with diarrhoea. C. jejuni was detected in mouse faeces throughout at levels of 10(7)-10(9) cfu/g. All mice killed whilst ill with diarrhoea displayed histopathological lesions typical of human campylobacteriosis. Severe pathology was limited to the large intestine and was suggestive of an acute, bacteria induced inflammation. Although blood was detected in the diarrhoeal stools, no evidence of mucosal epithelial cell invasion was found by immunohistology. No pathology was detected in tissue sections from any of the animals that had not developed signs of disease following C. jejuni inoculation. These immunodeficient mouse strains are readily, and heavily, colonised as adults by C. jejuni. The diarrhoea, although sporadic, was reproducibly produced, and could provide the basis for pathogenicity studies. PMID- 9736163 TI - Molecular probes for the detection of pathogenic fungi in the presence of human tissue. AB - Four primer systems, amplifying fragments of the gene coding for the small ribosomal subunit (18S rRNA) were characterised with pure cultures of 65 medically relevant fungal species plus two mushrooms. A primer cocktail (TR1/CA1 TR2/AF2) amplified 59 of 67 fungal species; the universal fungal primer 1 (UF1) in combination with the eukaryotic primers S3 or EU1 amplified 64 and 65 of 67 fungal species, respectively. The design of an additional primer (RZY1) enabled the amplification of the missing members of the zygomycetes. The primer systems amplified all the medically relevant fungi tested. These included eight Candida spp. and seven other yeast species, 13 dermatophytes, 32 moulds (including six zygomycetes and five dimorphic fungi) and two mushrooms. Eleven controls including DNA from Schistosoma mansoni, Escherichia coli, Mycobacterium tuberculosis and man were not amplified. The oligonucleotide CA hybridised with C. albicans, C. tropicalis and C. parapsilosis; the oligonucleotide TR hybridised with the 13 dermatophytes; the oligonucleotide AF hybridised with Aspergillus fumigatus, A. flavus, A. terreus, A. nidulans, A. versicolor, A. tamarii, A. clavatus, A. fischeri, but not with A. niger or A. versicolor; and the oligonucleotide HC hybridised with three varieties of Histoplasma capsulatum. These oligonucleotides did not hybridise with the other fungi nor the controls. The specificity of the newly designed primer systems was confirmed by selective amplification of fungal DNA from human lung tissue spiked with fungal biomass and from vitrectomy fluid of a patient with candida endophthalmitis. PMID- 9736164 TI - Clinical evaluation of the Mycobacteria Growth Indicator Tube (MGIT) compared with radiometric (Bactec) and solid media for isolation of Mycobacterium species. AB - The aim of this study was to evaluate the clinical use of a new culture system for the isolation of mycobacteria. Routine clinical specimens were cultured in the Mycobacteria Growth Indicator Tube, the radiometric Bactec 460 TB system and on Lowenstein Jensen (LJ) medium to compare recovery rates and times for detection of mycobacteria and contamination rates. MGIT was tested for its ability to support the growth of a wide range of mycobacterial species. Acid-fast bacilli (AFB) were detected on direct smears of 76 of 603 clinical specimens and mycobacteria were isolated by at least one method from 109 specimens; 93% of these were detected in the MGIT, 95% in the Bactec 460 TB system and 87% on LJ medium. The MGIT, Bactec and LJ media detected 92%, 97% and 95%, respectively, of 61 M. tuberculosis isolates and 94%, 94% and 77% of the 48 isolates belonging to the M. avium complex (MAC). The mean detection times in MGIT, Bactec and LJ media for M. tuberculosis were 22, 14 and 27 days respectively, and for MAC were 14, 12, and 29 days, respectively. Growth of M. tuberculosis was detected in Bactec, within 4 weeks, in 93% of the 61 culture-positive specimens, compared with only 61% in MGIT and 66% on LJ. The number of MAC detected within 4 weeks was similar in Bactec and MGIT, but less in LJ medium. Differences in sensitivity and time to detection of growth between media were greater for specimens in which AFB were not detected on direct smear than those on which AFB were seen. Contamination rates were similar in the three systems (3-4%). MGIT supported the growth of all 28 Mycobacterium spp. inoculated. MGIT has significant safety advantages and is less labour intensive than other methods, but the time to detection of M. tuberculosis, especially in smear-negative specimens, was longer in MGIT than in Bactec. PMID- 9736165 TI - Heterogeneous location of the mupA high-level mupirocin resistance gene in Staphylococcus aureus. AB - Epidemiologically unrelated clinical isolates of Staphylococcus aureus with high level resistance to mupirocin (MIC > or = 512 mg/L) were studied to determine the location of the mupA resistance gene. The gene was carried on plasmids of variable size, some of which were transferable in vitro. DNA hybridisation of genomic DNA from 85 isolates showed that mupA was located on EcoRI fragments of seven different sizes; the most frequently observed fragments were 7 kb (46 isolates) or 4.1 kb (21 isolates). All isolates retained a 1.6-kb Nco I fragment that hybridised with mupA probes, but showed heterogeneous hybridisation patterns after digestion with Hinc II. These data suggested that mupA may be conserved, but that variation occurs in the flanking DNA proximal to it. Amplification of spacer regions between mupA and closest proximal copy of IS257 yielded products of variable size and was consistent with the presence of IS257 in either orientation. It is proposed that IS257-mediated events are responsible for the heterogeneity observed. The location of mupA varied between epidemiologically unrelated isolates of the same strain, including isolates of EMRSA-16 -- one of the two predominant methicillin-resistant strains in UK hospitals at the present time -- and this correlated with variations in the digestion patterns of the mupirocin resistance plasmids. The variable location of mupA should be evaluated further as a potential epidemiological tool with which to monitor the spread of high-level mupirocin resistance in EMRSA-16 or other strains of S. aureus. PMID- 9736166 TI - Synergic antistaphylococcal properties of lactoferrin and lysozyme. AB - Staphylococcus epidermidis colonises a wide range of implanted prosthetic devices, but rarely contact lenses -- despite a similarity in material composition. A conceivable explanation for this anomaly is the action of the tear defences, including the constitutive proteins lactoferrin and lysozyme. Therefore this study investigated the effect of lactoferrin, lysozyme and serum on the growth of S. epidermidis isolates in artificial tear fluid. Whether supplemented with serum alone or serum with either apolactoferrin or lysozyme, this medium induced a similar, strain-variable effect. However, simultaneous addition of these proteins induced a greater bactericidal or bacteristatic effect. Of those strains killed by the concerted action of apolactoferrin and lysozyme, the absence of serum led to a further increase in the bactericidal effect, whereas strains displaying bacteriostasis were unaffected by serum. Iron saturation of lactoferrin reversed the antimicrobial synergy of apolactoferrin and lysozyme. These results show synergy between lactoferrin and lysozyme which is dependent on the iron limitation of lactoferrin. As a bactericidal mechanism, this synergy is augmented by serum, but bacteriostasis remains unaffected by serum supplemention. Thus, the combination of lysozyme and lactoferrin may partly explain the low level of contact lens colonisation by S. epidermidis in vivo. PMID- 9736167 TI - Is GBV-C (hepatitis G virus) an innocent bystander? PMID- 9736168 TI - Biochemical and histological features of hepatitis G virus infection. AB - To assess the biochemical and histological characteristics of hepatitis G virus (HGV) infection, we examined four patients who were infected with HGV only (HGV group), and compared them with 16 patients infected with both HGV and hepatitis C virus (HCV; HGV + HCV group) and 18 patients infected with HCV only (HCV group). Biochemical examination showed a significantly low level of serum alanine aminotransferase (ALT) in the HGV group, and that the gamma-glutamyl transpeptidase (gamma-GTP)/ALT ratio in the same group was significantly higher than in the other two groups. Although all three patient groups had a similar degree of liver fibrosis, both the degree of periportal inflammation and total histological activity index were significantly lower in the HGV group than in the other two groups. Fibrous enlargement of the portal tract without lymphoid infiltration and thin fibrous septa was characteristically observed in the HGV group. No significant difference was found between the HGV + HCV group and HCV group. Our results suggest that biochemical and histological changes in HGV infection are very mild and quite different from those of HCV infection. PMID- 9736169 TI - Effects of hepatitis G virus coinfection on severity of hepatitis C: relationship to risk factors and response to interferon treatment. AB - The aims of the present study were to identify characteristics that are more often associated with hepatitis G virus (HGV) coinfection in Australian patients infected with the hepatitis C virus (HCV) and to investigate the effects of HGV on the histological and functional severity of chronic hepatitis C. Serum samples from 209 patients with chronic hepatitis C were tested for HGV-RNA using single round reverse transcriptase-polymerase chain reaction to primers directed at the NS5 region of the HGV genome. Hepatitis G virus RNA was detected in 40 cases (19%). Hepatitis G virus-coinfected patients tended to be younger and parenteral risks could be identified in all but six. Although country of birth did not differ significantly between the coinfected and HCV-alone groups, HGV-positive patients appeared to be less likely to have originated from Asia. On logistic regression analysis, HCV genotype 3a was found in a significantly higher proportion of patients with HGV coinfection than other genotypes (P < 0.01). Liver histology and response to interferon were similar in the HGV-coinfected and HCV-alone groups and liver-related complications appeared to occur less frequently in patients with both HGV and HCV. On univariate analysis, antipyrine clearance was found to be higher in the coinfected group (P < 0.05), implying better preservation of hepatic metabolic function, but this difference was lost when adjusted for HCV genotype. In conclusion, coinfection with HGV was more commonly associated with HCV genotype 3a, a genotype associated with injection drug use in younger patients. However, the presence of HGV coinfection did not adversely affect liver disease or the response to interferon treatment in patients with chronic hepatitis C. PMID- 9736170 TI - Efficacy of hypothermic perfusion using University of Wisconsin solution in extended hepatectomy with hepatic inflow occlusion in a canine model. AB - This study was designed to elucidate the efficacy of University of Wisconsin (UW) solution for preventing liver injury, when used as a hypothermic perfusate infused into the systemic circulation during extended hepatectomy with hepatic inflow occlusion. Adult mongrel dogs (9.5-17.5 kg, n = 14) were subjected to 75% hepatectomy under 60 min hepatic inflow occlusion. The animals were divided into two groups. The UW group (n = 7) underwent hypothermic perfusion using 4 degrees C UW solution (core temperature of the liver: 12.3 +/- 0.2 degrees C). The control group designated as the Ringer's lactate (LR) group (n = 7) underwent hypothermic perfusion using 4 degrees C LR solution. The perfusate was introduced into the systemic circulation via the hepatic vein. Blood from the hepatic vein was sampled, and alanine aminotransferase, purine nucleoside phosphorylase activities and the ammonia concentration were measured. The 7 day survival rate was higher in the UW group than in the LR group. The parameters of liver function were less significantly altered in the UW group than in the LR group. The plasma ammonia concentration was significantly (P < 0.05) lower 6 h after reperfusion in the UW group than in the LR group. A small volume of hypothermic perfusion of the liver using UW solution was safe if it returned to systemic circulation. Hypothermic perfusion of the liver using UW solution may be effective for preventing hepatic tissue injury during extended hepatectomy with hepatic vascular occlusion. PMID- 9736171 TI - Hepatocyte apoptosis and hepatic expression of transforming growth factor-beta1 mRNA during involution of hyperplastic rat liver induced by hepatocyte growth factor. AB - Hepatocyte apoptosis occurs during involution of hyperplastic liver induced by administration of xenobiotic compounds in rats. With this hyperplasia and involution, hepatic transforming growth factor (TGF)-beta1 is reported to be expressed to stimulate hepatocyte apoptosis. In regenerating liver after partial resection showing no hyperplasia, such expression of TGF-beta1 is also seen. However, no hepatocyte apoptosis develops despite the high levels of TGF-beta1. When rats received an intravenous injection of human hepatocyte growth factor at 12 h intervals for 14 days, the hepatic DNA content was increased 12 h after the last injection to 140% of control. This DNA content was significantly decreased at 108 and 180 h after discontinuation of treatment. At 60 h after the last injection, the number of apoptotic bodies positive for nick end-labelling of DNA in hepatocytes was significantly greater in treated rats than in control rats. Hepatocyte apoptosis was also identified electron micrographically. Hepatic TGF beta1 mRNA levels in treated rats were significantly lower than in control rats at 12 h and then gradually increased towards control levels. We conclude that hyperplastic liver induced in normal rats by hepatocyte growth factor regresses with hepatocyte apoptosis and suppressed hepatic TGF-beta1 mRNA levels. PMID- 9736172 TI - A comparative study of nucleolar organizer region, proliferating cell nuclear antigen and epidermal growth factor receptor staining in colon tumours. AB - Several studies have reported that proliferating cell nuclear antigen (PCNA), nucleolar organizer regions (NOR) and epidermal growth factor receptor (EGF-R) are cellular proliferation parameters in malignant tumours. However, the correlation of EGF-R with PCNA and NOR has not been reported in colon tumours. To clarify this, we investigated the correlation of the expression of EGF-R with PCNA and NOR in colonic epithelial tumours. Using immunohistochemical staining and one-step silver staining techniques, the expressions of EGF-R-, PCNA- and NOR associated protein were studied in paraffin sections from five normal mucosae, 10 adenomas with mild atypia, 16 adenomas with moderate atypia, 32 adenomas with severe atypia including carcinoma in situ and 47 with invasive colon carcinomas. Significant differences were found among the respective degrees of atypia in Ag NOR counts, PCNA labelling index (LI), and the expression of EGF-R. However, there were no significant differences among the Ag-NOR counts of the severe atypia and invasive carcinoma. There was a significant positive correlation between Ag-NOR counts and PCNA LI (P < 0.001). The Ag-NOR counts and PCNA LI in EGF-R-positive cases were higher than in EGF-R-negative cases (P < 0.001). These results indicate that each of the parameters increased significantly with the increase in atypia and the progression of cancer. Epidermal growth factor receptor provokes an exacerbation of protein synthesis and the cell cycle in colonic epithelial tumours. PMID- 9736173 TI - A quantitative immunohistochemical evaluation of inflammatory cells at the affected and unaffected sites of inflammatory bowel disease. AB - Levels of T lymphocytes, histiocytes and mast cells have been reported to be increased in the affected mucosa of Crohn's disease (CD) and ulcerative colitis (UC), but the colorectal distribution of these cells is not fully understood. We hypothesized that differences in cell densities between CD and UC would be characteristic, not only in the affected, but also in the unaffected mucosa. The aims of the present study were to clarify whether there were any differences in cell densities in CD, UC and infectious colitis (IC) in the affected mucosa and between CD and UC in the unaffected mucosa. Using mouse monoclonal antibodies recognizing memory T cells (OPD4), cytotoxic/suppressor T cells (C8/144B), histiocytes (PG-M1) and mast cells (AA1), we evaluated mucosal cell densities in biopsy specimens from both endoscopically affected and unaffected sites of CD (n = 12) and UC (n = 15) and from affected sites of IC (n = 10). Ten normal controls were also examined. At affected sites, all cells were significantly more abundant in UC than in the other conditions, except that the density of PG-M1+ in CD was similar to that seen in UC. Although the densities of OPD4+ and C8/144B+ cells at unaffected sites were slightly higher in both CD and UC and in UC, respectively, there was no significant difference in cell densities between CD and UC. The ratio of OPD4+ cell density at affected sites to that at unaffected sites was appreciably higher in UC than in CD. The results suggest that a common feature of UC and CD is an increase in PG-M1+ cells at the affected mucosa but that the other inflammatory cells studied are more abundant, particularly in UC, and that the difference between UC and CD is conspicuous when comparing the OPD4+ cell density of the affected mucosa with that of the unaffected mucosa. PMID- 9736174 TI - Geranylgeranylacetone, an anti-ulcer drug, stimulates hexosamine production in a rat gastric mucosal cell line through binding to a specific cytosolic protein. AB - An anti-ulcer drug, geranylgeranylacetone (GGA), stimulates hexosamine production in a rat gastric mucosal cell line (RGM-1). The aim of this study was to elucidate the mechanism of this action. The role of protein kinase A, inositol phospholipid turnover and tyrosine kinase in the stimulatory action of GGA on hexosamine production in RGM-1 was determined by observing cAMP production, [3H] inositol phosphate turnover and western blotting of tyrosine phosphorylation, respectively. Any trophic effect of GGA on RGM-1 was also checked by [3H] thymidine incorporation. Our experiments showed that GGA has no effect on cAMP production, inositol phospholipid turnover, tyrosine phosphorylation or DNA synthesis in RGM-1. Finally, a [14C]-GGA competitive receptor binding assay was performed on RGM-1 and we found that [14C]-GGA specifically bound to RGM-1 cytosolic protein. Although retinoic acid (RA), another polyisoprenoid compound significantly stimulated hexosamine production in RGM-1, we confirmed that the [14C]-GGA binding site in RGM-1 is different from the RA binding site. In summary, GGA stimulates hexosamine production in RGM-1 and this action is probably mediated through its binding to a specific cytosolic protein in RGM-1. PMID- 9736175 TI - Assessment of autonomic nervous activity during gastrointestinal endoscopy: analysis of blood pressure variability by tonometry. AB - We continuously measured blood pressure by tonometry in 30 patients during endoscopy to determine the influence of upper gastrointestinal endoscopy on cardiac events. Patients were divided into two groups: one group treated with scopolamine butylbromide as premedication (SB group) and another group without premedication (C group). Time- and frequency domain analyses of beat-to-beat systolic blood pressure variability were performed for 128 consecutive beats. For time-domain analysis, we calculated the coefficient of variation of systolic blood pressure (CV(BP)). For the frequency domain analysis, we determined the low frequency (LF(BP); 0.04-0.15 Hz) and high-frequency (HF(BP); 0.15-0.40 Hz) powers of the variation in systolic blood pressure and the ratio of LF(BP) to HF(BP) (LF(BP)/HF(BP)) during endoscopy. The CV(BP) and HF(BP), indicators of parasympathetic tone, increased in the early phase of endoscopy but decreased significantly in the middle and late phases compared with the pre-endoscopy value. The ratio of LF(BP)/HF(BP), an indicator of indirect sympathetic tone, increased throughout the endoscopic procedure. Moreover, premedication with scopolamine butylbromide prevents the excessive parasympathetic nervous reflex when an endoscope passes through the upper digestive tract and also brings both decreased parasympathetic tone and increased sympathetic tone at the late phase of endoscopic procedure. Our results indicate that gastrointestinal endoscopy induced an autonomic nervous abnormality, which may contribute to the occurrence of cardiac events during endoscopic procedures. PMID- 9736176 TI - Severe cholestatic jaundice induced by Epstein-Barr virus infection in the elderly. AB - Infectious mononucleosis due to Epstein-Barr virus (EBV) is almost always a self limited disease, most commonly seen in young adults. Hepatitis is a well recognized complication of EBV infection that usually resolves spontaneously. Jaundice occasionally results from the unusual complication of autoimmune haemolytic anaemia rather than hepatitis. We report a 60-year-old man with severe cholestatic jaundice whose history, liver histology and laboratory findings suggested EBV infection. He also developed significant jaundice related to his hepatitis, but not to autoimmune haemolysis, a situation that led to diagnostic delay. Costly diagnostic laboratory tests and invasive procedures were performed to rule out a malignant extrahepatic biliary obstruction. Physicians need to be aware of this complication and EBV infection should be included in the differential diagnosis of cholestatic jaundice in the elderly. PMID- 9736177 TI - Three paediatric cases of primary sclerosing cholangitis treated with ursodeoxycholic acid and sulphasalazine. AB - We present here three paediatric patients with primary sclerosing cholangitis. In case 1, the serum gamma-glutamyl transpeptidase was decreased only temporarily by ursodeoxycholic acid (UDCA) treatment and 34 months later, sulphasalazine was added because of microscopic colitis. The enzyme level decreased with dual therapy. Similarly, in case 3, first diagnosed as autoimmune hepatitis, the transpeptidase levels remained elevated for 18 months during treatment with UDCA, prednisolone and mizoribin. The enzyme decreased only after a diagnosis of primary sclerosing cholangitis complicated with ulcerative colitis was established and sulphasalazine was introduced. Case 2 also had Crohn's colitis and was put on UDCA and sulphasalazine from the start. The enzyme level was normalized within 1 month and has remained normal for the following 5 years. Liver biopsies were analysed repeatedly in these three patients. In case 1, periductal fibrosis remained unchanged while being treated by UDCA. There appeared to be no progression in liver cirrhosis in case 3 while being treated by UDCA, prednisolone, and mizoribin. In case 2, who has been treated with both UDCA and sulphasalazine from the start, periductal fibrosis and portal fibrosis were remarkably improved 45 months later. We suggest that sulphasalazine in addition to UDCA might be a viable treatment for children with primary sclerosing cholangitis. PMID- 9736178 TI - Candida oesophagitis. PMID- 9736179 TI - Hepatobiliary and pancreatic imaging. Eventration of the right diaphragm and acute cholecystitis. PMID- 9736180 TI - Hepatobiliary tuberculosis. AB - Tuberculosis is known to involve the liver in different ways. The term hepatobiliary tuberculosis refers to the localized form of hepatic tuberculosis as a distinct clinical entity, with signs and symptoms related to the hepatobiliary tract. Its clinical features and the different diagnostic aids used in its diagnosis are reviewed. Plain abdominal radiographs showing diffuse hepatic calcifications seen in approximately 50% of cases are almost diagnostic for hepatobiliary tuberculosis. Liver biopsies obtained either by ultrasound, computed tomography or laparoscopy, showing caseating granuloma usually establish the diagnosis. In the absence of caseation necrosis, a positive acid-fast bacillus (AFB) or culture for Mycobacterium tuberculosis is needed to establish the diagnosis. A polymerase chain reaction assay for the identification of Mycobacterium tuberculosis in liver biopsy specimens is a new development. Treatment is similar to that used for pulmonary tuberculosis. Quadruple therapy (using four anti-tuberculosis drugs) is recommended, generally for 1 year. For patients with obstructive jaundice, in addition to anti-tuberculous treatment, biliary decompression should be performed either by stent insertion during endoscopic retrograde cholangiopancreatology, by percutaneous transhepatic biliary drainage or by surgical decompression whenever feasible. PMID- 9736181 TI - Isolation and primary culture of rat Kupffer cells. AB - The aim of this study was to describe a reproducible method for the isolation, purification and primary culture of rat Kupffer cells. Kupffer cells were isolated following sequential pronase/collagenase digestion of the liver and enrichment of a non-parenchymal cell fraction by a single-density gradient centrifugation step using 30% metrizamide. Kupffer cells were isolated and further purified from this cell fraction by centrifugal elutriation. Kupffer cells were isolated at 1017 g at 48-110 mL/min. All Kupffer cell fractions exhibited phagocytosis of 3 microm latex beads. Kupffer cell fractions isolated at 48 and 60 mL/min were predominantly ED2 negative while later fractions (80-110 mL/min) were ED2 positive. Kupffer cells were adherent in culture after 2 h. This method for Kupffer cell isolation resulted in a yield of 80-120 x 10(6) Kupffer cells per liver. PMID- 9736183 TI - Culture of sinusoidal endothelial cells from rat liver. PMID- 9736182 TI - Isolation and culture of rat hepatic stellate cells. PMID- 9736184 TI - Culture and transfection of mammalian primary hepatocytes and hepatocyte-derived cell lines. PMID- 9736185 TI - Introduction: why protect the human genome? PMID- 9736187 TI - What's special about molecular genetic diagnostics? AB - In its first part, this paper seeks to make plausible (a) that molecular genetic diagnostics differs in ethically relevant ways from traditional types of medical diagnostics and (b) that the consequences of introducing this technology in broad screening-programs to detect widespread genetic diseases in a population which is not at high risk may change our understanding of health and disease in a problematic way. In its second part, the paper discusses some aspects of public control of scientific and technological innovations in the field of molecular genetic diagnostics. PMID- 9736186 TI - A map to a new treasure island: the human genome and the concept of common heritage. AB - While the 1970's have been called the environmental years, the 1990's could be seen as the genome years. As the challenge to map and to sequence the human genome mobilized the scientific community, risks and benefits of information and uses that would derive from this project have also raised ethical issues at the international level. The particular interest of the 1997 UNESCO Declaration relies on the fact that it emphasizes both the scientific importance of genetics and the appropriate reinforcement of human rights in this area. It considers the human genome, at least symbolically, as the common heritage of humanity. PMID- 9736188 TI - Genetic screening and ethics: European perspectives. AB - Analysis and comparison of genetic screening programs shows that the extent of development of programs varies widely across Europe. Regional variations are due not only to genetic disease patterns but also reflect the novelty of genetic services. In most countries, the focus for genetic screening programs has been pregnant women and newborn children. Newborn children are screened only for disorders which are treatable. Prenatal screening when provided is for conditions for which termination may be offered. The only population screening programs for adults are those for thalassaemia carrier status in Cyprus, Greece and Italy. Social responses to genetic screening range from acceptance to hostility. There is a fundamental tension between individual and community in the debates in various European countries about implementation of screening programs. Opposition to genetic screening is frequently expressed in terms of arguments about "eugenics" with insufficient regard to the meaning of the term and its implications. Only a few countries have introduced explicit legislation on genetic screening. Legislation to address discrimination may provide more safeguards than legislation protecting genetic information itself. PMID- 9736189 TI - Geneticization: the Cyprus paradigm. AB - Geneticization is a broad term referring to several related processes such as a spreading tendency to use a genetic model of disease explanation, a growing influence of genetics in medical practice, and the slow changing of individual and societal attitudes towards reproduction, prevention and control of disease. These processes can be demonstrated in medical literature on preventive genetic screening and counselling programs for beta-thalassaemia in Cyprus, the United Kingdom and Canada. The preventive possibilities of the new genetic and diagnostic technologies have been quickly understood and advocated by health professionals, and their educational strategies have created a web of social control, in marked contrast to the alleged voluntary decision-making process and free choice. Genetic diagnostic technologies have led to considerable changes in control and management of beta-thalassaemia, and have generated a number of unresolved incongruities. PMID- 9736190 TI - Genotyping in clinical trials: towards a principle of informed request. AB - This paper reviews the usefulness of bioethical instruments such as the informed consent principle to handle ethical and political challenges of clinical trials in genotyping and DNA-banking and discusses an informed request model as well as other contractual relations between research institutions, patients, and their families. PMID- 9736191 TI - Public health and bioethics. AB - Conduct that satisfies certain bioethical doctrines may come into conflict with the needs and ethics of public health. The growth of antibiotic resistance in bacteria and the spread of HIV both contribute to the difficulty of controlling infectious disease. These two sets of priorities need to be reconciled and this is likely to require a reassessment of prevailing ethical doctrines in the face of the needs of public health. PMID- 9736192 TI - Interventions in the human genome: some moral and ethical considerations. AB - In the debate regarding the different possibilities for gene therapy, it is presupposed that the manipulations are limited to the nuclear genome (nDNA). Given recent advances in genetics, mitochondrial genome (mtDNA) and diseases must be considered as well. In this paper, we propose a three dimensional framework for the ethical debate of gene therapy where we add the genomic type (nDNA vs. mtDNA) as a third dimension to be considered beside the paradigmatic dimensions of target cell (somatic vs. germ-line) and purpose (therapeutic vs. enhancement). Somatic gene therapy can be viewed today as generally accepted, and we review the contemporary arguments surrounding it on the basis of bioethical-pragmatic, socio political and deontological classifications. Many of the supposed ethical questions of somatic gene therapy today are not new; they are well-known issues of research ethics. We also critically summarize the different international perspectives and the German ethical discussion regarding manipulations of germ line cells. PMID- 9736193 TI - The patentability of human genes: an ethical debate in the European Community. AB - The European Parliament rejected in 1995 the European Commission proposal to harmonize legal protection of biotechnological inventions. Although it did not seem initially the most contentious of the many issues involved in the current legal and ethical debate around biomedicine and genetics, patenting is now focusing bioethics in Europe. PMID- 9736194 TI - Discussing HUGO: the German debate on the ethical implications of the Human Genome Project. AB - The current German criticism of HUGO centers around the term 'human dignity'; consenquentialist and autonomy-based arguments are used. The debate culminates in questioning the integrity of bioethics as a scholarly discipline and has created a heterogeneous coalition of disparate political and social groups that oppose any research that would facilitate genetic pre-selection of human characteristics. PMID- 9736195 TI - UNESCO: universal declaration on the human genome and human rights. PMID- 9736196 TI - Transient cortical blindness in preeclampsia with indication of generalized vascular endothelial damage. AB - A 26-year-old woman with twin fetuses of 28 weeks' gestational age had symptoms of preeclampsia and was admitted to the hospital for observation. Nine days later, after reporting a severe headache, the patient experienced loss of vision in both eyes. An emergency computed tomographic brain scan was performed to rule out intracranial hemorrhage, and cesarean delivery was performed. Twelve hours after the operation, the patient's vision improved gradually and returned to normal after 24 hours. The placenta was submitted to pathologic examination, and magnetic resonance imaging was performed 4 days after birth. Recent thrombosis observed in the histologic section of the placenta, ischemic changes in the brain seen in the computed tomographic and magnetic resonance scans, and severe proteinuria manifested clinically suggest vascular endothelial damage as the underlying mechanism in this case of preeclampsia-related transient cortical blindness. PMID- 9736197 TI - Nonarteritic anterior ischemic optic neuropathy with macular edema: visual improvement and fluorescein angiographic characteristics. AB - The objective of the study was to assess visual outcomes in patients who have nonarteritic anterior ischemic optic neuropathy (NAION) with macular edema (ME). Thirteen eyes (12 patients) with NAION and ME were observed for an average of 5.3 months after onset of visual loss. Intravenous fluorescein angiography was performed on 10 eyes. Fluorescein leakage was observed in 8 of 10 (80%) eyes. Leakage was nonfocal, minimal, and diffuse. Eleven of 13 (85%) eyes with ME improved. The average improvement was 2.3 Snellen lines. The presence of ME in patients with NAION may confer a better visual prognosis than reported in patients with NAION alone. PMID- 9736198 TI - Anterior ischemic optic neuropathy in a disc with a cup: an exception to the rule. PMID- 9736199 TI - Comparison of cholinergic supersensitivity in third nerve palsy and Adie's syndrome. AB - OBJECTIVE: To determine whether the degree of cholinergic supersensitivity of the pupil differs in patients with preganglionic injury of the oculomotor nerve (third nerve palsy) compared with patients with postganglionic injury (Adie's pupil). METHODS: In this retrospective study, the authors first identified 11 patients with oculomotor nerve palsy and 11 patients with unilateral Adie's pupil who demonstrated supersensitive pupillary responses using dilute pilocarpine. The same methods for testing supersensitivity of the iris sphincter, and for defining its presence, had been used in both groups of patients. Pupil diameters of the affected and unaffected fellow eye were measured directly from self-developing photographs obtained before and 30 minutes after pilocarpine 0.1% was applied to both eyes. The amount of absolute constriction of the affected pupil, as well as the net constriction of the affected pupil (i.e., the amount of pilocarpine induced constriction of the unaffected pupil subtracted from the amount of pilocarpine-induced constriction of the affected pupil), was compared between the two groups of patients using the Mann-Whitney test. RESULTS: No significant differences were identified in any of the comparisons. CONCLUSIONS: The degree of cholinergic supersensitivity of the iris sphincter appears to be similar regardless of whether the site of injury along the parasympathetic pathway of the oculomotor nerve is preganglionic or postganglionic. PMID- 9736200 TI - Elevation of serum tumor marker CA 15.3 levels as the first manifestation of metastatic breast cancer to the cavernous sinus. AB - We report rising tumor marker levels of CA 15.3 as the presenting manifestation of metastatic breast cancer to the cavernous sinus and orbit. A 39-year-old woman with a history of breast cancer developed increasing levels of tumor marker CA15.3. Ten months later, she developed vision loss in the right eye, diplopia, and right-sided ptosis. A magnetic resonance scan of the head showed a mass involving the right cavernous sinus and superior orbital fissure. Biopsy of the lesion showed metastatic breast cancer. She was treated with surgery and radiotherapy and did well. Ophthalmologists should be aware of the significance of increasing levels of tumor markers, such as CA 15.3, in patients with a history of breast cancer and new neuroophthalmologic signs or symptoms. PMID- 9736201 TI - Paraneoplastic optic neuropathy and autoantibody production in small-cell carcinoma of the lung. AB - A 59-year-old woman presented with acute-onset, bilateral, painless loss of vision, dysarthria, and ataxia. Ophthalmoscopy showed bilateral optic disc edema. A magnetic resonance scan of the head was normal. Chest radiography showed mediastinal adenopathy. Mediastinoscopy and biopsy identified small-cell carcinoma of the lung. An autoantibody to optic nerve and retina was demonstrated in the patient's serum. An electroretinogram was normal. The patient was diagnosed with a paraneoplastic optic neuropathy and paraneoplastic cerebellar syndrome. After treatment for her lung cancer, the patient remains stable from a visual and neurologic standpoint. PMID- 9736202 TI - Autonomic nervous system involvement in Behcet's disease: a pupillometric study. AB - The aim of this study was to elucidate whether autonomic nervous system dysfunction exists in patients with Behcet's disease by pupillometric tests. Thirty-one patients with Behcet's disease with a mean age of 41.3 years (range 21 64) and 41 control subjects with a mean age of 39.5 years (range 18-66) were selected for the study. To test the autonomic nervous system, four pupillometric techniques were used: pupil cycle time (PCT), dark-adapted pupil size (DAPS), 0.05% pilocarpine drop test, and 1% phenylephrine drop test. In all four tests, there were significant differences between the patients and controls. Mean PCTs were 1,156 ms (range 856-1,560 ms) and 919 ms (range 650-1,261 ms) in the patients and controls, respectively (p < 0.0001). The mean DAPS was 0.45 (range 0.31-0.66) in the patients, whereas it was 0.56 (range 0.42-0.67) in controls (p < 0.001). Iris sensitivity to both 0.05% pilocarpine and 1% phenylephrine showed significant differences between patients and controls, respectively (p < 0.05, p < 0.05). Among all four tests, only 0.05% pilocarpine sensitivity was correlated with the duration of Behcet's disease (p < 0.05). The results suggest that the autonomic nervous system innervating the iris is affected in Behcet's disease. This involvement may be due to the vasculitic nature of Behcet's disease. PMID- 9736203 TI - Intracranial Ewing's sarcoma. AB - Three patients with intracranial Ewing's sarcoma had neuro-ophthalmologic manifestations. In one patient, the primary tumor was in the skull and in two, it involved the long bones. Two patients complained of intermittent headache associated with bilateral, transient visual symptoms suggestive of migraine, which prompted imaging that showed occipital metastases. The third patient had an orbital syndrome. PMID- 9736204 TI - Microbial keratitis as the manifestation of trigeminal amyloidoma at initial presentation. PMID- 9736205 TI - Pachymeningitis with multiple cranial neuropathies and unilateral optic neuropathy secondary to Pseudomonas aeruginosa: case report and review. AB - Hypertrophic cranial pachymeningitis is a rare disorder that frequently presents with multiple cranial neuropathies. This disorder, which is characterized by thickening and infiltration of the cranial dura, can result from a variety of inflammatory and infectious conditions. A patient with hypertrophic cranial pachymeningitis is described in whom meningeal biopsy and bacterial cultures of the biopsy specimen revealed Pseudomonas aeruginosa. The authors believe this to be the first documented case of pachymeningitis secondary to this organism. PMID- 9736206 TI - Optic disk tubercle. AB - The purpose of this case report was to present a rare case of optic disk tubercle. The optic disk edema resolved on antituberculous therapy with recovery of vision. We concluded that visual loss from an optic disk tubercle can be the presenting sign of tuberculosis. PMID- 9736207 TI - Isolated fourth nerve palsy from midbrain hemorrhage: case report. PMID- 9736208 TI - Anterior visual pathway meningiomas primarily resected between 1978 and 1988: the Mayo Clinic Rochester experience. AB - The relapse rate, overall survival, and factors associated with a decreased recurrence-free survival rate in patients with anterior visual pathway (AVP) meningioma were compared with these features in patients who had meningiomas at other sites. Management of these patients is discussed. A review of the records of 581 consecutive patients who had primary resection of meningiomas between 1978 and 1988 identified 43 patients with AVP meningioma. Multiple clinical, surgical, and pathologic factors at the initial examination were analyzed to assess their association with recurrence, and the patients who had AVP meningioma were compared with patients who had non-AVP meningiomas to determine the factors that may influence recurrence. Recurrence-free and overall survival rates were determined. The AVP tumors were associated with a higher rate of recurrence. Subtotal resection was more common in the AVP tumors, but it alone was not associated with the decrease in recurrence-free or overall survival rates. Several factors that may explain the higher recurrence rate in patients with AVP meningioma were identified. Anterior visual pathway meningioma is associated with a higher rate of recurrence than are meningiomas at other sites. Operation remains the mainstay of treatment for symptomatic nonseeing eyes. Radiation therapy seems to be effective for managing recurrent tumor. PMID- 9736209 TI - Ocular motility review 1996. PMID- 9736210 TI - Alfred Bielschowsky's 1940 legacy for neuro-ophthalmology. AB - OBJECTIVES: The author was stimulated to write this article by a 1996 visit to the University where Professor Alfred Bielschowsky was Chairman of Ophthalmology in the 1930s. Dr. Bielschowsky was one of the founders of neuro-ophthalmology. This review, with biographical notes, is presented in his honor. Dr. Bielschowsky and the author had similar disruptive experiences, of historic interest, during the Hitler regime in Nazi Germany. REVIEW: Professor Bielschowsky's legacy begins with his contributions to ocular physiology. For instance, his after-image test establishes the presence of retinal correspondence, important for stereoscopic vision. Alfred Bielschowsky taught how an ocular examination is critical for neuro-ophthalmologic diagnosis, localization, prognostication, and treatment. Much of our knowledge is linked with his name. Examples include "Bielschowsky's Phenomenon", explaining dissociated vertical movements, and "Bielschowsky's Doll's Head Phenomenon" (Doll's Eyes), describing proprioceptive reflexes important for localizing intracranial lesions. Dr. Bielschowsky emphasized many pitfalls in the differential diagnosis of oculomotor anomalies. For example, he cautioned against mistaking the compensatory head position in congenital fourth cranial nerve paresis for neck muscle disease. CONCLUSION: Dr. Bielschowsky's emphasis on the clinical examination remains critical despite today's advanced diagnostic equipment. His legacy is the application of physiology to patient care. PMID- 9736211 TI - Idiopathic thrombocytopenic purpura (ITP): a historical odyssey. AB - Purpura has been recognized since ancient times and its clinical syndromes were refined by important observations in the sixteenth, seventeenth and eighteenth centuries. It required the development of adequate microscopes in the nineteenth century, however, to recognize the platelet, leading to the recognition of the thrombocytopenic component of ITP. The twentieth century brought recognition of the pathophysiology of the disorder and further defined the clinical states and treatments for ITP. The latter half of the twentieth century has focussed on the autoimmune components of ITP, initially on the humoral immune aspects and more recently on dysregulation of cellular immunity. PMID- 9736212 TI - The Mpl ligand and platelet homeostasis. AB - The classification of Mpl as a cytokine receptor present on cells of the platelet lineage has led to the identification and cloning of its ligand. This has resulted in a rapid accumulation of data advancing the understanding of the processes of megakaryopoiesis and thrombopoiesis and the regulation of endogenous Mpl ligand (thrombopoietin, eTPO). Highlights of in vitro human and non-human primate data will be discussed, as well as preclinical (in vivo) non-human primate studies. Two recombinant forms of Mpl ligands (rTPO) are currently being tested in clinical trials and early results will be reviewed. The preclinical and clinical studies will be summarized with consideration of the observations which provide insights into the biology of the response to exogenous rTPO. Understanding the biology of platelet production and the condition of target cells in treatment populations will facilitate the appropriate use of this potential therapeutic agent. PMID- 9736213 TI - Mechanisms for induction of autoimmunity in humans. AB - The triggering or immunogenic stimulus for human autoimmune diseases is unknown. It is not even known whether the stimulus is endogenous, i.e. truly "self" or exogenous, "non-self". Many autoimmune diseases are human lymphocyte antigen (HLA)-associated and demonstrate linkage to the HLA chromosome in family investigations. For most of these diseases, evidence is strong that the association is directly dependent on specific HLA class I or II molecules rather than on other genes located in the HLA region. Since HLA polymorphic HLA molecules have so far only been shown to have two distinct functions, both of which are immunological, a HLA association supports the notion that a particular disease is autoimmune. Furthermore, an association to a specific HLA allele implies that the immunogenic stimulus for autoimmunity would be one specific HLA binding peptide and that, at least initially, autoimmunity is dependent on the reactivity of one or a limited number of potentially autoaggressive T cell clones. These findings are encouraging and formin the basis for future preventive measures. One current theory is that autoimmune disease is precipitated by an environmental agent, such as a viral infection. Several different mechanisms to explain how a viral infection could induce autoimmune disease in humans are described and one specific example is presented for a virus-induced autoimmune disease in humans. The question of whether ITP could also be dependent on such a mechanism is briefly discussed. PMID- 9736214 TI - Analysis of transmembrane signalling and T cell defects associated with idiopathic thrombocytopenic purpura (ITP). AB - Adult chronic idiopathic thrombocytopenic purpura (ITP) is an autoimmune disease characterized by production of autoreactive antibodies to platelet antigens. It is now becoming clear that autoantibody production, in general, is regulated by T helper (Th) cells. Several recent studies have examined potential defects in T cell function in this disease and have demonstrated that patients with ITP possess abnormal lymphocyte activation and Th1/Th2-mediated cytokine production. Although the underlying cause(s) of aberrant T cell function in this disease are not known, studies from other models of autoimmune disease indicate that defects in T cell transmembrane signalling can be causally linked to abnormal T cell activation and cytokine production. This review will present some of the major T cell signalling pathways and discuss how altered T cell signalling may be linked to autoimmunity with an emphasis on ITP. Recent preliminary findings of a potential defect in the signal transduction apparatus in lymphocytes from three patients with ITP will also be presented. PMID- 9736216 TI - Role of complement in immune or idiopathic thrombocytopenic purpura. AB - The clinical course of immune or idiopathic thrombocytopenic purpura (ITP) is variable, suggesting different mechanisms for the decreased platelet count. The complement factors C3 and C4 have been detected on platelets, both alone and in association with immunoglobulin G (IgG), and a reduced platelet survival time has been described. Platelets have the capacity to interact with the complement system since they have both complement receptors and complement regulatory proteins on their cell membranes. The membrane attack complex (C5b-9) induced by antiplatelet antibodies generates platelet microparticles in a concentration dependent manner. A marked variation in resistance to this phenomenon has been demonstrated between individuals and between men and women. These platelet microparticles seem to retain their biological role in haemostasis. Platelets also appear to play a role in the processing of immune complexes. Immunoglobulins and complement factors are found in several clinical situations where circulating immune complexes are expected. Furthermore, human platelets bind immune complexes in vitro and the reaction can be blocked by antireceptor antibodies to immunoglobulins and complement. These findings raise a number of questions about the role of complement in the pathophysiology of ITP. PMID- 9736215 TI - Platelet proteins as autoantibody targets in idiopathic thrombocytopenic purpura. AB - Idiopathic thrombocytopenic purpura (ITP), caused by autoantibodies directed against certain platelet antigens, is the most common entity of the immune thrombocytopenias. ITP is an acquired disorder and can affect both children and adults. However, the clinical syndromes of ITP are distinct between children and adults. Childhood (acute) ITP characteristically is acute in onset, occurs within 1-2 weeks of an infection, usually of viral origin, resolves spontaneously within 6 months. Adult (chronic) ITP has an insidious onset and rarely resolves spontaneously. Over the last decade considerable new information has accumulated as to the pathophysiological mechanisms of immune thrombocytopenias. In addition, most of the knowledge on this disorder has been obtained from studies of adult patients with chronic ITP. The present work gives an updated overview of the platelet autoantigens and the molecular immunological reactions in ITP. PMID- 9736217 TI - Immunobiology of T helper cells and antigen-presenting cells in autoimmune thrombocytopenic purpura (ITP). AB - Autoimmune thrombocytopenic purpura (AITP) is a bleeding disease in which autoantibodies are directed against the individual's own platelets, resulting in enhanced Fc-mediated platelet destruction by macrophages in the reticuloendothelial system. Most research in AITP has focused on characterization of the autoantibodies, while little has been devoted to the cellular immune mechanisms leading to autoantibody production. This report summarizes the current state of the literature and argues that enhanced T helper cell/antigen-presenting cell interactions in patients with AITP are the primary stimulus for the development of antiplatelet autoantibody production. Understanding these events is important for eventually identifying disease-initiating platelet autoantigens and ultimately developing specific immunotherapies for AITP. PMID- 9736218 TI - Role of the T cell receptor in idiopathic thrombocytopenic purpura (ITP). AB - During the past few decades a number of studies has described T cell defects and attempted to elucidate their role in the pathogenesis of idiopathic thrombocytopenic purpura (ITP). Some studies implicate T cells as potential initiators of autoantibody production in ITP. However, only a few of these have studied the role that the T cell receptor may play in the pathogenesis of ITP. In a variety of autoimmune syndromes interest has focused on the alpha- and beta chains of the T cell receptor. Deviations from the normal T cell receptor gene usage have been reported in rheumatoid arthritis, systemic lupus erythaematosus and multiple sclerosis. Usually, these studies have shown a restricted heterogeneity of T cell receptor variable gene usage. The studies on the T cell receptor in ITP have included a limited number of patients, which makes it difficult to evaluate the significance of the role that the T cell receptor may play in the pathogenesis of ITP. Further studies are warranted. PMID- 9736219 TI - Measurement of platelet antibodies: where do we stand? AB - The immune pathogenesis of idiopathic thrombocytopenic purpura (ITP) is well established; however, the diagnostic criteria do not require identification of platelet autoantibodies. The abundance of methods devised for the measurement of platelet antibodies over the years provides evidence of the so-far futile search for a reliable diagnostic assay. The latest generation of platelet antigen specific assays has widened the knowledge of autoimmune mechanisms in thrombocytopenia, but they still lack sufficient sensitivity to be a useful tool in the routine evaluation of thrombocytopenic patients. PMID- 9736220 TI - Mechanisms in childhood idiopathic thrombocytopenic purpura (ITP). AB - The concepts of the pathological mechanisms in childhood idiopathic thrombocytopenic purpura (ITP) have, to a great extent, been based on clinical experience and on data generated in adults. Studies performed in children have demonstrated that platelet antigen-specific autoantibodies are present in chronic ITP and, to a lesser extent, in acute ITP. It is, however, likely that the mechanisms initiating the production of autoantibodies are different in the two entities. In acute ITP, production of autoantibodies and immune complexes is probably linked to a transient antiviral immune response. Chronic ITP in children is an autoimmune process which eventually is reversible in many cases. The initiating factors, as in other autoimmune disorders, are yet to be elucidated. PMID- 9736221 TI - Platelet function in autoimmune (idiopathic) thrombocytopenic purpura. AB - Platelets play an essential role in the formation of haemostatic plugs. The quantitative defect of platelets in autoimmune (idiopathic) thrombocytopenic purpura (ITP) can result in bleeding complications, but most ITP patients have platelets with normal or enhanced function. Platelets in ITP are large, young, so called "stress" platelets with increased platelet-associated autoimmune antibody (immunoglobulin G). Young stress platelets are more functional platelets, and their presence may account for bleeding times in ITP patients that are shorter than would be predicted on the basis of the patients' (low) platelet counts. Some ITP patients have significant mucocutaneous bleeding with platelet counts >50 x 10(9) l(-1); this may be due to qualitative platelet dysfunction (e.g. brought about by inhibitory antiplatelet autoantibodies). PMID- 9736222 TI - Cytokines in idiopathic thrombocytopenic purpura (ITP). AB - Most research in idiopathic thrombocytopenic purpura (ITP) has focused on characterization of the autoantibodies directed against platelet antigens resulting in enhanced platelet elimination by macrophages. This report summarizes the current knowledge of cytokine pattern found in individuals with ITP. Serum assessment has demonstrated increased levels of interleukin (IL)-2 and interferon gamma (IFN-gamma), while IL-4 was significantly decreased. In addition, thrombopoietin (TPO) has been found in normal levels while IL-11 has been reported to be elevated. These data indicate that ITP is associated with a Th1 type of T helper cytokine response, while that of type Th2 is downregulated. Initially, megakaryocytes are found at normal levels in bone-marrow aspirates, explaining the unchanged production of TPO. The increase in IL-11 may be reflected by the increased number of platelets being produced per megakaryocyte. However, there is little information on these events in immunocompetent sites such as bone marrow, spleen and lymph nodes. PMID- 9736223 TI - Reticulated platelet counts in the assessment of thrombocytopenic disorders. AB - Reticulated platelets (RP) are the youngest platelets in the circulation and can be measured by analysing the RNA content of platelets from whole blood or platelet-rich plasma by flow cytometry. Increased RP are indicative of increased production of platelets. Despite the current lack of standardization for the measurement of RP, it is useful in the assessment of patients with ITP by aiding the distinction of these patients from those with decreased platelet production. RP counts also have a role in the assessment of the complicated patient with multiple possible aetiologies for thrombocytopenia. Measurement of the RP count may hold predictive value for marrow recovery following myelosuppressive or myeloablative chemotherapy, and may play a role in monitoring the administration of the various thrombopoietins currently under clinical trial. PMID- 9736224 TI - Short-course oral prednisone therapy in children presenting with acute immune thrombocytopenic purpura (ITP). AB - Immune thrombocytopenic purpura (ITP) is a disorder for which management remains controversial. The ongoing goal is to define the minimal therapy required for children with acute ITP. A pilot study of short-course oral prednisone (4 mg(-1) kg(-1) d(-1) for 4 d with no tapering) was undertaken in 25 consecutive children with acute ITP and platelet counts under 20 x 10(9) l(-1). Of the 25 children, 22 responded to the prednisone therapy by achieving a platelet count higher than 20 x 10(9) l(-1) within 1 week of commencing treatment. This regimen was found to be safe, inexpensive and effective in increasing the platelet count of children to a haemostatically safe level. PMID- 9736225 TI - Current management issues of childhood and adult immune thrombocytopenic purpura (ITP). AB - The management of acute and chronic immune thrombocytopenic purpura (ITP) of children differs in many aspects from that of adults. Current paediatric and adult treatment options are discussed in this review in the light of the recently published practice guidelines for the diagnosis and treatment of ITP issued by a panel of paediatric and adult haematologists on behalf of the American Society of Hematology. Uncontrolled rather than controlled randomized studies often represent the basis for treatment decisions. Important issues in improving the management of patients with ITP include the identification of research priorities resulting in controlled clinical trials with well-defined study endpoints, the logistics and coordination of research activities and their presentation at international meetings. PMID- 9736226 TI - Directions for research in autoimmune thrombocytopenic purpura (ITP). AB - All attendees participated in a round-table discussion regarding directions for research in autoimmune thrombocytopenic purpura (ITP). Suggested areas for study were grouped into five main areas: (i) improved classification of ITP identifying subsets of patients with differing clinical syndromes and response to treatment, and those more likely to have serious bleeding manifestations; identification of patients with reduced thrombopoiesis was emphasized; (ii) studies aimed at elucidating the aetiology and pathophysiology of ITP, with emphasis on distinctions between acute and chronic ITP and between patients responsive or refractory to therapy; these studies focused on measures of humoral and cellular immune dysregulation; (iii) studies of platelet function in ITP, with the intent of defining these abnormalities and correlating them with the clinical manifestations of the disease; (iv) new approaches to treatment, particularly of refractory patients; and (v) a miscellaneous group, which included development of an ITP registry, evaluation of the "burden" of disease, investigation of mood changes in ITP, etc. The discussion was not intended to be all-inclusive, but focused on the content of other talks in this symposium. It is hoped that some of these suggestions will be further developed for investigation in multicentre co operative studies to improve the diagnosis, understanding and treatment of ITP. PMID- 9736227 TI - Are elevated cerebrospinal fluid levels of IL-6 in sudden unexplained deaths, infectious deaths and deaths due to heart/lung disease in infants and children due to hypoxia? AB - Many SIDS cases probably die after periods of hypoxia and it has been shown that hypoxia may stimulate IL-6 production. The purpose of this paper was to examine if there were any correlations between hypoxanthine in vitreous humour and IL-6 in CSF. The concentration of IL-6, IL-1beta and TNFalpha in cerebrospinal fluid of 50 Sudden Infant Death syndrome (SIDS) cases, 9 borderline SIDS cases, 18 infectious deaths, 8 violent deaths and 22 cases with heart/lung disease were measured by ELISA. The hypoxanthine (Hx) vitreous humour concentrations in the same groups were determined by high performance liquid chromatography. The IL-6 levels in cases of infectious death, heart/lung disease and borderline cases were significantly higher than in the SIDS cases (p < 0.01). The Hx levels were in the same range in cases of SIDS, borderline SIDS and infectious death, and they were significantly higher than the levels in cases of violent death and heart/lung disease (p < 0.01). There was no correlation between hypoxanthine and IL-6 in any of the groups. In the cases studied IL-6 elevation is probably not induced by hypoxia, but is rather a result of immunological stimulation. PMID- 9736228 TI - Plasma levels of carboxy terminal propeptide of type I procollagen and pyridinoline cross-linked telopeptide of type I collagen in healthy school children. AB - The aim of this study was to reveal the relationship of the plasma levels of the carboxy terminal propeptide of type I procollagen (PICP) and the pyridinoline cross-linked carboxyterminal telopeptide domains of type I collagen (ICTP) to age and height velocity (HV), and to compare PICP and ICTP levels in those who have not reached their final height with those who have. PICP and ICTP levels were measured by RIA in 271 healthy children (161M, 110F) aged from 10 to 15 y. The HV was calculated from their health check-up cards. The subjects were divided into two groups in this study: the final height (FH) group whose HV was <1.0 cm/y, and the non-final height (NFH) group whose HV in the last year was > or =1.0 cm/y. PICP and ICTP levels almost paralleled the values of age-related changes of HV. Furthermore PICP and ICTP levels significantly correlated with HV. PICP and ICTP levels of the FH group were higher than those of adults previously reported. The values will be useful as reference for healthy children and growth disorder. Before reaching final height, PICP and ICTP can be useful makers of not only bone turnover but also of linear growth. Bone turnover rate is still increasingly active just after linear growth has been completed, and then it become similar to the levels of adults. PMID- 9736229 TI - The prevalence and related symptomatology of Helicobacter pylori in children with recurrent abdominal pain. AB - The aim of the study was to assess and compare the IgG seroprevalence of H. pylori in children with recurrent abdominal pain with healthy children and to investigate the related symptoms. IgG antibodies against low-molecular weight H. pylori antigens were assessed in 438 children with recurrent abdominal pain and in 91 healthy controls. Sera with an ELISA unit-value above the cut-off level were confirmed by Western immunoblot. Only seropositive children with recurrent abdominal pain were examined by an oesophago-gastro-duodenoscopy. Symptomatology was recorded according to the localization of the abdominal pain, presence of pyrosis, nocturnal pain, relation of pain to meals and bowel irregularities. The seroprevalence was 21% (95% CI: 17-25%) in the children with recurrent abdominal pain and 10% (95% CI: 5-18%) in the healthy controls (p = 0.30). In seropositive children with RAP H. pylori was found in 46/66 by culture and histology. The presence of H. pylori was significantly associated with active or inactive chronic gastritis. The presence of H. pylori was associated with both parents being born in a country with a high prevalence and a low social class. Helicobacter pylori-positive children had more often pain related to meals than the H. pylori-negative children. No differences among the two groups were seen according to the levels of haemoglobin, leucocytes, thrombocytes, weight and height. In conclusion, the seroprevalence of H. pylori is comparable in children with recurrent abdominal pain and healthy children. No specific symptomatology was seen in H. pylori-positive children with RAP. PMID- 9736231 TI - Importance of training for correct Turbuhaler use in preschool children. AB - AIM: To study the impact of both audiovisual information and nurse-training on the use of budesonide Turbuhaler in preschool children who had never used a dry powder inhaler. DESIGN: A single-blind, randomized, parallel-group trial studied 72 children aged 3-5 y. All children and their parents were shown an instructional video and given written instruction. After this, peak inspiratory flow (PIF1) through Turbuhaler was measured. Children in group A (n = 36) then received individual training by a nurse while those in Group B (n = 36) did not and PIF2 was measured. Afterwards, Group B received similar individual training while Group A received no additional training, and PIF3 was measured. Group A was given a placebo Turbuhaler and encouraged to practice at home. Two weeks later, both groups returned to the clinic where PIF4 was measured. RESULTS: The number of children who were able to correctly perform PIF1, PIF2 and PIF4 in Group A was 27, 34 and 36, respectively. The corresponding numbers for Group B were 30, 29, and 29. No effect of training was seen in 3-y-old children. Individual training by a nurse was associated with a statistically significant increase in PIF2 (10 l/min; p = 0.014). Moreover, 2 weeks of home training was associated with an additional increase in PIF of 8 l/min compared with Group B (p < 0.015). After individual instruction and home training, mean PIF in children aged 4 and 5 was 56 (42-72) and 55 (41-66) l/min, respectively. CONCLUSION: After individual instruction and training at home, the vast majority of children aged 4 and 5 y can use Turbuhaler correctly. Audiovisual information and individual instruction is not sufficient in the majority of these children. Few 3-y-old children can learn the correct use of Turbuhaler. PMID- 9736230 TI - The intestinal ecosystem in chronic functional constipation. AB - Chronic functional constipation is common in infants, and the bacterial composition of stools in this condition is not known. The study aims were to: (i) investigate the composition of the intestinal ecosystem in chronic functional constipation; (ii) establish whether the addition of the water-holding agent calcium polycarbophil to the diet induces an improvement in constipation; and (iii) determine the composition of the intestinal ecosystem after the use of this agent. In total, 42 children (20F, 22M; mean age: 8.6 +/- 2.9 y) were studied. Twenty-eight children with functional chronic constipation without anatomical disorders were treated double-blind in random sequence for 1 month with an oral preparation of calcium polycarbophil (0.62 g/twice daily) or placebo. Intestinal flora composition was evaluated by standard microbiological methods and biochemical assays on faecal samples collected before and after treatment. Fourteen healthy children were studied as controls. The results show that (i) the constipated children presented a significant increase in clostridia and bifidobacteria in faeces compared to healthy subjects--different species of clostridia and enterobacteriaceae were frequently isolated; no generalized overgrowth was observed; Clostridia outnumbered bacteroides and E. coli mean counts by 2-3log, while bacteroides and E. coli counts were similar (5-6 log10/g fresh faeces); these intestinal disturbances could be defined as a dysbiosis, i.e. a quantitative alteration in the relative proportions of certain intestinal bacterial species. (ii) Clinical resolution of constipation was achieved only in 43% of treated children and an improvement in 21% (one bowel movement every 2 d). (iii) Calcium polycarbophil treatment induced no significant changes in the composition of the intestinal ecosystem, nor in blood chemistry parameters. PMID- 9736232 TI - Erythrocyte superoxide dismutase activity and plasma malondialdehyde levels in children with Henoch Schonlein purpura. AB - Reactive oxygen molecules (ROM) have been suggested to contribute to many pathological conditions including vasculitides and renal diseases. In the present study we measured the activity of superoxide dismutase (SOD) as an antioxidant enzyme in red blood cells and the level of malondialdehyde (MDA), which is a product and an indicator of lipid peroxidation, in the plasma of 16 children (7M, 9F) with Henoch Schonlein purpura (HSP) at the onset of the disease (SOD 1 and MDA 1) and at the remission period (SOD 2 and MDA 2). The results were compared with the results of 17 healthy children studied as a control group. There was no significant difference for SOD activities between the patients in each period and the control group (p > 0.05). There was a statistically significant difference between MDA 1 and MDA 2 levels (p < 0.01), each of which were also significantly different from the MDA levels of control group (p < 0.001 and p < 0.01, respectively). The effect of ROMs on different clinical conditions of HSP was also examined and lipid peroxidation was found to be increased more in patients with renal involvement. It is concluded that oxidant stress especially lipid peroxidation plays an important role in the pathogenesis of HSP and in development of renal injury. PMID- 9736233 TI - A missense mutation, Val62Ala, in the glucokinase gene in a Norwegian family with maturity-onset diabetes of the young. AB - Maturity-onset diabetes of the young (MODY) is a form of diabetes mellitus characterized by autosomal dominant inheritance, onset usually before 25 y of age and a primary defect in glucose-stimulated insulin secretion. It is a heterogeneous disorder both with respect to aetiology and clinical features. Mutations in the genes encoding the glycolytic enzyme glucokinase, the liver enriched transcription factors, hepatocyte nuclear factor-1alpha (HNF-1alpha), HNF-1beta and HNF-4alpha, and the transcription factor, insulin promoter factor-1 (IPF-1) have all been associated with MODY. Here, we report a family, Norway-2 (N2), characterized by the presence of a mild, complication-free form of diabetes with autosomal dominant inheritance. Sequencing of the glucokinase gene in the proband revealed a T-to-C mutation in codon 62 which resulted in a valine-to alanine substitution, designated Va162Ala (V62A). The V62A mutation, which has not been previously reported, cosegregated with diabetes in the N2 family. The results presented here indicate that the glucokinase form of MODY occurs in Norway. Moreover, screening the glucokinase gene for mutations in other families with clinical features similar to those of the N2 family could lead to improved treatment for patients with this form of diabetes. PMID- 9736234 TI - Metabolic control in children with insulin-dependent diabetes mellitus 5 y after diagnosis. Early detection of patients at risk for poor metabolic control. AB - Children (n = 38) aged 3-15 y were randomly chosen, at the time of diabetes diagnosis, for conventional management at a hospital ward, or for treatment partly in a training apartment where the family was offered problem-based education and special therapeutic support. HbA1c, blood glucose stability, urinary C-peptide excretions and incidence of hypoglycaemic attacks and diabetes ketoacidosis (DKA) were monitored and some standardized, self-estimated psychological tests were performed during the first 2 y after diagnosis. During the 3 y thereafter, HbA1c, presence of DKA, microalbuminuria, retinopathy and hypertension were monitored. None of the patients demonstrated signs of diabetes microangiopathy or DKA. The overall mean HbA1c level was 7.2% 5 y after diagnosis and 30% of the children had HbA1c values <6.3%. There were no differences in the HbA1c values for the patients treated by the different management regimens. Blood glucose variability (SD) was also similar, with 75% of the values in the range of 3-10 mmol/l. Patients with poor glycaemic control (mean HbA1c >8.3%) year 5 after diagnosis had already the second year after diagnosis significantly higher HbA1c values and blood glucose variability. The fathers of these patients demonstrated a higher degree of maladjustment. On the basis of increasing HbA1c values, high blood glucose variability and psychosocial risk factors such as their fathers' emotional responses, patients at risk for poor metabolic control in the future can be identified within 2 y after diagnosis. Efforts and resources can thus be focused at an early stage on this group. PMID- 9736235 TI - Quality of life of young adults with idiopathic short stature: effect of growth hormone treatment. Dutch Growth Hormone Working Group. AB - The aim of the study was to evaluate whether treatment with recombinant human growth hormone (rhGH) affects the quality of life of young adults who were diagnosed as idiopathic short stature (ISS) during childhood, and whether their quality of life and aspects of the personality are different from normal. Experiences and expectations concerning rhGH treatment of the subjects and their parents were also investigated. Eighty-nine subjects were included into the study: 24 subjects (16M, 8F) were treated with rhGH from childhood, whereas 65 subjects (40M, 25F) were never treated. At the time of the interview all subjects had attained final height [mean (SD) -2.3 (0.9) SDS for Dutch references], and the age of the treated subjects was 20.5 (1.0) y, and 25.7 (3.5) y of the control subjects (p < 0.001). The level of education was similar, but the treated subjects had less often a partner compared to the control subjects (adjusted for age and gender, p < 0.001). The Nottingham Health Profile and Short Form 36 Health Survey showed no difference in general health state between treated and control subjects, and the healthy Dutch age-specific references (norm group). Although 74% of the subjects reported one or more negative events related to their height, and 61% would like to be taller, only 22% and 11% were willing to trade-off at Time Trade-Off and Standard Gamble, respectively. The personality of the subjects, which was measured by the Minnesota Multiphasic Personality Inventory, was not different from the norm group. The satisfaction with the rhGH treatment was high, as it had caused 12 (8) cm and 13 (7) cm gain in final height according to the subjects and parents, respectively. Based on initial predicted adult height (Bayley & Pinneau), this gain was only 3.3 (5.6) cm. We concluded that although the treated subjects had a partner less often when compared to the control subjects, the quality of life of subjects with ISS at adult age is normal and appears not to be affected by rhGH therapy, The treated subjects were very satisfied with the treatment, probably by overestimation of the final height gain. PMID- 9736236 TI - Sarcoidosis in children. Epidemiology in Danes, clinical features, diagnosis, treatment and prognosis. AB - This paper reviews current knowledge of childhood sarcoidosis with regard to the epidemiology in Danes, clinical presentation, diagnostic procedures, treatment and prognosis. Sarcoidosis is a granulomatous disease of unknown aetiology, with multiorgan involvement. The diagnosis is confirmed by the demonstration of epitheloid cell granulomas in tissue biopsy specimens. During the period 1980-92, three cases of childhood sarcoidosis were recorded in Copenhagen County, which has a total population of 610,000. The approximate incidence of clinically recognized sarcoidosis in Danish children younger than 15 y of age was 0.22 0.27/100,000 children per year, corresponding to approximately three new cases in Denmark each year. The true incidence is unknown, since the disease is often asymptomatic and resolves without a clinical diagnosis being made. In children younger than 5 y of age, the disease is characterized by involvement of skin, eyes and joints, whereas in older children involvement of lungs, lymph nodes and eyes predominate. The mainstay of treatment consists of oral corticosteroids. The risk/benefit ratio of using long-term corticosteroids needs to be evaluated in each individual patient. Some patients may benefit from additional therapy with methotrexate. The long-term prognosis is not well established, but it seems to be poorer in children younger than 5 y. Older children appear to have as favourable a prognosis as young adults. PMID- 9736237 TI - Clinical profiles of subjects with subcortical leukomalacia and border-zone infarction revealed by MR. AB - Clinical profiles of 13 children with subcortical leukomalacia and border-zone infarction revealed by MR were analysed. The causes of brain damage were neonatal asphyxia, hypoglycaemia, circulatory disturbance associated with congenital heart diseases, and other perinatal events in five, three, two and three, respectively. Severe, moderate and mild mental retardation were present in three, five and five, respectively. Of the seven children who could walk alone, six had ataxia and one spastic diplegia; the remaining six manifested truncal instability. The ataxia was deduced to have resulted from cerebral lesions. PMID- 9736238 TI - Failure of short-term mannose therapy of patients with carbohydrate-deficient glycoprotein syndrome type 1A. AB - Carbohydrate-deficient glycoprotein syndrome type 1A (CDGS1A) is an inherited disorder with multisystemic abnormalities resulting from failure to generate sufficient lipid-linked oligosaccharide precursor or to transfer the sugar chain to many glycoproteins. Cultured fibroblasts from these patients have reduced incorporation of mannose into glycoproteins which can be corrected by adding D mannose to the culture medium. Providing dietary mannose to elevate mannose concentrations in vivo therefore might remedy some of the underglycosylation in the patients. Five children with CDGS1A aged 15 months to 14 y completed a protocol of enteral supplementation with D-mannose 100 mg/kg every 3 h for 9 d. The mean S-mannose level increased from 32 microM (range 22-42 microM) to a trough value of 72 microM (range 39-103 microM). No serious side effects were observed. Surprisingly, the mean serum concentration of four glycoproteins (transferrin, alpha1-antitrypsin, antithrombin, and thyroxine-binding globulin) tended to decrease, and the mean serum concentration of carbohydrate-deficient transferrin (CDT) increased. Furthermore, the initially present abnormal isoforms of these glycoproteins and of protein C became more prominent and/or additional abnormal isoforms appeared. This short-term trial does not support a benefit of mannose to the deficient glycosylation of CDGS1A patients. PMID- 9736239 TI - Cerebral excitatory amino acids and Na+,K+-ATPase activity during resuscitation of severely hypoxic newborn piglets. AB - We tested the hypothesis that early brain recovery in hypoxic newborn piglets is improved by resuscitating with an O2 supply close to the minimum level required by the newborn piglet brain. Severely hypoxic 2-5-d-old anaesthetized piglets were randomly divided into three resuscitation groups: hypoxaemic (n = 8), 21% O2 (n = 8), and 100% O2 groups (n = 8). The hypoxaemic group was mechanically ventilated with 12-18% O2 adjusted to achieve a cerebral venous O2 saturation of 17-23% (baseline; 45 +/- 1%, mean +/- SEM). During the 2h resuscitation period, extracellular aspartate and glutamate concentrations in the cerebral striatum were higher during hypoxaemic resuscitation (p = 0.044 and p = 0.055, respectively) than during resuscitation with 21% O2 or 100% O2, suggesting an unfavourable accumulation of potent excitotoxins during hypoxaemic resuscitation. The cell membrane Na+,K+-ATPase activity of cerebral cortical tissue after 2 h resuscitation was similar in the three groups (p = 0.30). In conclusion, hypoxaemic resuscitation did not normalize early cerebral metabolic recovery as efficiently as resuscitation with 21% O2 or 100% O2. Resuscitation with 21% O2 was as efficient as resuscitation with 100% O2 in this newborn piglet hypoxia model. PMID- 9736240 TI - Treatment of extremely premature newborns: a survey of attitudes among Danish physicians. AB - In order to describe the attitudes towards the treatment of extremely preterm infants, a questionnaire presenting a series of fictitious situations concerning imminent extremely preterm labour and treatment of an infant born after 24 weeks gestation was mailed to all physicians employed at obstetrical or paediatric departments in Denmark. The questionnaire was designed in two versions, differing as regards the parents' situation and attitude towards treatment. Each version was sent to half of the sample. Of 954 questionnaires 664 (69.6%) were completed and returned. Most respondents advocated active treatment prior to and immediately upon delivery, but many would withhold more intensive treatments or withdraw treatments in case of severe complications. The parents' situation and attitude towards treatment played a role in forming the decision choice to a significant proportion of the respondents. In severe cases, many would provide morphine in doses that could unintentionally hasten death, while few were in favour of legalizing active euthanasia. PMID- 9736241 TI - Nephropathy and hypertension as manifestations in a 13-y-old girl with primary antiphospholipid syndrome. AB - Severe renal hypertension due to both unilateral renal arterial occlusion and renal thrombotic microangiopathy developed in a 13-y-old girl as a manifestation of primary antiphospholipid antibody syndrome. The combination of the intravenous high-dose urokinase therapy and oral anticoagulation therapy, comprising aspirin, warfarin and dipyridamole, was significantly effective in improving her renal function and preventing thrombotic events during an 18-month follow-up period. PMID- 9736242 TI - A child with bruising. PMID- 9736243 TI - Vascular hamartoma: another cause of testicular torsion in a newborn. PMID- 9736244 TI - Development and differentiation of endothelium. AB - Vascular endothelial cells play an important role in tissue homeostasis, fibrinolysis, and coagulation; blood-tissue exchange, vasotonus regulation, blood cell activation, and migration; and the vascularization of tissues. The formation of new blood vessels comprises two distinct steps: vasculogenesis, the in situ assembly of capillaries, and angiogenesis, the sprouting of capillaries from preexisting ones. Vascular endothelial growth factor (VEGF) is essential for vasculogenesis and angiogenesis. Its expression is high in the embryonic brain and kidney when angiogenesis occurs and low in the adult brain when angiogenesis is absent. In the kidney, VEGF expression remains high in glomerular podocytes even in the adult. VEGF receptors 1 and 2 (fit-1 and flk-1) are endothelial specific receptor tyrosine kinases. Similar to the ligand, expression of these receptors is high during brain and kidney angiogenesis, low in adult brain endothelium, but high in adult glomerular endothelium. Because VEGF is also a vascular permeability factor, the expression in the adult correlates with the low permeability of blood-brain barrier endothelium and the high permeability of fenestrated glomerular endothelium. Although fenestrae formation can be induced in vitro by VEGF and a basal lamina-type extracellular matrix, blood-brain barrier characteristics seem to require the presence of still unknown brain derived factors. PMID- 9736245 TI - Origins and formation of microvasculature in the developing kidney. AB - Regulation of microvessel assembly in the developing kidney is not known and may occur through vasculogenic, angiogenic, or both processes. To examine this question, we grafted rat and mice embryonic (E) day 12 (E12) kidneys, which have only a rudimentary vasculature, into anterior eye chambers of mouse and rat hosts. Species-specific, monoclonal anti-basement membrane antibodies showed that glomerular basement membranes, mesangial matrices, and microvessel basement membranes were always derived from the graft. When wild-type E12 mouse kidneys were grafted into anterior chambers of ROSA26 mice, in which the beta galactosidase transgene is expressed ubiquitously, glomerular and microvascular endothelial cells stemmed from the graft, even after maintenance of kidneys in organ culture for 6 days before grafting. Immunolabeling with antibodies against the vascular endothelial growth factor (VEGF) receptor, Flk1, the EphB1 receptor, and its ligand, ephrin-B1, labeled discrete mesenchymal cells in embryonic and newborn kidney cortex, as well as developing microvessel and glomerular endothelium. In adult kidneys, Flk1 labeled glomeruli weakly, other vascular structures were unlabeled. When wild-type E12 kidneys were grafted under renal capsules of adult ROSA26 hosts, endothelium developing within the graft again came from the implanted kidney. In contrast, when E12 kidneys were grafted into renal cortices of newborns, glomeruli within grafts now contained host-derived endothelium. Similarly, when ROSA26 E12 kidneys were implanted into newborn wild type hosts, chimeric vessels containing graft- and host-derived endothelium were seen in nearby host tissue. Our results indicate that cells capable of forming the entire microvascular tree of grafted metanephroi are already present in the E12 kidney. We hypothesize that Flk1/VEGF and EphB1/ephrin-B1 mediate renal endothelial mitosis-motility and cell guidance-aggregation behavior, respectively. PMID- 9736246 TI - Role of angiotensin in renal vascular development. AB - All components of the renin-angiotensin system (RAS) are expressed in the developing kidney in a temporospatial pattern that suggests a role for this system in kidney morphogenesis. Pharmacological blockade of angiotensin actions in fetal and newborn animals results in striking alterations in kidney architecture, including immature glomeruli and papillae, dilated tubuli, and arrested vascular development. Inactivation of angiotensinogen or angiotensin converting enzyme genes in mice results in similar anomalies that begin as subtle alterations in early life and become more pronounced as extrauterine life progresses. However, inactivation of each angiotensin receptor subtype does not result in obvious morphological abnormalities, suggesting functional redundancy at the receptor level. Crossing of mice lacking the various receptor subtypes should be revealing. Overall, the available information suggests that the RAS is necessary for the normal morphological and functional development of the kidney and the preservation of kidney architecture in adult life. PMID- 9736247 TI - Heterogeneous activation mechanisms in the renal microvasculature. AB - Vascular smooth muscle cells in different renal microvascular segments utilize different activation mechanisms to respond to mechanical and vasoactive stimuli. L-type Ca2+ channel blockers vasodilate primarily the preglomerular vascular resistance component responsible for autoregulation. Local interstitial infiltration of Ca2+ channel blockers increases glomerular pressure and markedly reduces vascular responsiveness of the tubuloglomerular feedback mechanism. Ca2+ channel blockers selectively attenuate the afferent vasoconstrictor responses to increases in perfusion pressure. Although both afferent and efferent arterioles constrict in response to angiotensin II (Ang II), afferent but not efferent constriction requires Ca2+ influx through L-type Ca2+ channels. Sensitivity of the preglomerular arterioles to Ang II is also heterogeneous with the greatest sensitivity in glomerulus-near, terminal segments. Adenosine triphosphate (ATP) is a vasoconstrictor agonist that selectively activates Ca2+ entry pathways in afferent arterioles but has no effect on efferent arterioles. In isolated preglomerular smooth muscle cells, increasing extracellular [KCl] increases intracellular Ca2+ by stimulating voltage-dependent Ca2+ influx. Ang II, norepinephrine, and ATP also elicit similar increases in intracellular Ca2+. Mechanical and agonist-induced voltage-dependent Ca2+ influx is thus a primary pathway in the control of cytosolic Ca2+ in afferent arterioles. Efferent arterioles, however, rely primarily on intracellular Ca2+ mobilization and other Ca2+ influx pathways. PMID- 9736248 TI - Localization and interactions of vasoactive peptide receptors in renomedullary interstitial cells of the kidney. AB - Vasoactive peptides regulate renal medullary microcirculation and tubular function, but the localization of their receptors and mechanisms of actions are currently unknown. Using electron microscopic autoradiography, we have mapped the receptors for angiotensin II (Ang II [AT1 and AT2]), endothelin (ET(A) and ET(B)), and bradykinin (B2) in the rat renal medulla. Although these peptide receptors show distinct vascular and tubular distributions, they overlap strikingly in renomedullary interstitial cells (RMICs) of the inner stripe and the papilla. Using reverse transcription-polymerase chain reaction (RT-PCR) and Southern analysis, mRNAs for AT1A, ET(A), and B2 receptors were detected in cultured adult RMICs. Ang II increases intracellular inositol 1,4,5-triphosphate (IP3) and [Ca2+]i and stimulates [3H]thymidine incorporation and extracellular matrix (ECM) synthesis via AT1A receptors. Endothelin and bradykinin also stimulate cell proliferation and ECM synthesis in RMICs through ET(A) and B2 receptors, respectively, but the actions of endothelin are modulated by concurrent nitric oxide production. By contrast, AT2 receptor mRNA was detected only in embryonic RMICs, in which Ang II inhibits cell proliferation through this receptor. These results suggest that multiple vasoactive peptides may interact with RMICs to exert endocrine and/or paracrine influences on renal medullary microcirculation and tubular function. PMID- 9736249 TI - Structural and molecular dissection of the juxtaglomerular apparatus: new aspects for the role of nitric oxide. AB - The juxtaglomerular apparatus (JGA) is composed of the macula densa (MD), the extraglomerular mesangium, and the juxtaglomerular arterioles. The JGA functions to adapt glomerular filtration rate (GFR) to distal tubular [NaCl] and to adjust the synthesis and release of renin. The type 1 isoform of nitric oxide synthase (NOS1) is present in MD cells, and release of NO toward the glomerular vasculature is thought to modulate signaling at the JGA. Chronic alterations in GFR and/or tubular [NaCl] are paralleled by adjustments of NOS1. Molecular characterization of NOS1 mRNA reveals several renal variants suggesting cell type specific regulation at the level of transcription and translation. PMID- 9736250 TI - Identification of ceramide targets in interleukin-1 and tumor necrosis factor alpha signaling in mesangial cells. AB - An increasing number of cell-surface receptors have been shown to trigger sphingomyelin turnover and generation of the lipid signaling molecule ceramide. Ceramide plays a role in mediating cellular responses as diverse as inflammation, differentiation, gene expression, growth suppression, and apoptosis. A radioiodinated, photoaffinity-labeling analog of ceramide ([125I]TID-ceramide) was used to identify downstream signaling targets of ceramide. Ceramide was found to bind specifically to and activate protein kinase c-Raf, leading to subsequent activation of the extracellular signal-regulated kinase (ERK) module in mesangial cells. We found also that ceramide binds to and differentially modulates the activity of distinct protein kinase C isoenzymes. These data are discussed in the context of interleukin 1beta-induced inflammatory gene expression in mesangial cells. PMID- 9736251 TI - Tubuloglomerular feedback: new concepts and developments. AB - Luminal [NaCl] at the macula densa (MD) has two established effects: regulation of glomerular arteriolar resistance through tubuloglomerular feedback (TGF) and control of renin secretion. TGF acts as a minute-to-minute stabilizer of distal salt delivery, thereby minimizing the impact of random perturbations in filtration and absorption forces on NaCl excretion. During long-lasting perturbations of MD [NaCl], control of renin secretion becomes the dominant function of the MD. The potentially maladaptive effect of TGF under chronic conditions is prevented by TGF adaptations permitting adjustments in glomerular filtration rate to occur. TGF adaptation is mechanistically coupled to the endpoint targeted by chronic deviations in MD [NaCl], the rate of local and systemic angiotensin II generation. Studies of TGF in transgenic mice are expected to provide further insights into the mechanisms mediating between luminal [NaCl] and afferent arterioles. TGF responses are virtually abolished in mice in which either the AT1A gene or the angiotensin converting enzyme gene is rendered nonfunctional by homologous recombination. In contrast, TGF responses are unaltered in nitric oxide synthase I knockout mice. Thus, an intact renin angiotensin system appears to be critical for the TGF signaling pathway. PMID- 9736252 TI - Characteristics of isolated perfused juxtaglomerular apparatus. AB - Tubuloglomerular feedback (TGF), which operates between the tubule and the parent glomerulus, is important to renal autoregulation and homeostasis of body fluid and electrolytes. The juxtaglomerular apparatus (JGA) has long been suggested as the anatomical site of TGF. To study the function of the JGA directly, we developed an in vitro preparation in which both the afferent arteriole (Af-Art) and macula densa (MD) of a microdissected rabbit JGA are microperfused simultaneously. We see that increasing the [NaCl] of the MD perfusate constricts the afferent arteriole in the segment close to the glomerulus. This constriction is blocked by furosemide, a loop diuretic known to inhibit TGF. On the other hand, microperfusion of Af-Arts alone showed the myogenic response to exist in the more proximal segments. Such an anatomical relationship between the myogenic response and TGF may enable the kidney to achieve its extremely efficient autoregulation. PMID- 9736253 TI - Prostaglandins in macula densa function. AB - Cyclooxygenase (COX)-2 mRNA and immunoreactive protein localize to the macula densa and adjacent cortical thick ascending limb in renal cortex, and chronic NaCl restriction increases expression of this enzyme. These findings suggest an integral role for eicosanoids generated by macula densa-associated COX-2 in mediating renin release. As selective inhibitors of COX-2 become available, it will be important to assess their effects on the renin-angiotensin system and glomerular hemodynamics. PMID- 9736254 TI - Macula densa nitric oxide synthase: expression, regulation, and function. AB - The type 1 brain nitric oxide synthase (bNOS) isoform occurs in macula densa (MD) cells where it functions to vasodilate the afferent arteriole and blunt expression of tubuloglomerular feedback (TGF). Dietary salt restriction enhances bNOS expression, yet microperfusion studies with NOS inhibitors imply that it is functionally inactive. We thus assessed the hypothesis that reduced L-arginine (L Arg) availability during low salt (LS) intake limits MD NO generation. Maximal TGF responses were recorded during Henle's loop perfusion with artificial tubular fluid (ATF). Microperfusion of L-Arg into the MD of LS, but not normal or high salt (HS) rats blunted maximal TGF responses (8.0 +/- 0.4 to 6.0 +/- 0.5 mm Hg; N = 23; P < 0.01). Response to L-Arg was stereospecific, inhibited by coperfusion with monomethyL-L-arginine (L-NMA), and dependent on system y+ transport, because it was blocked by coperfusion with the competitors L-lysine or L-homoarginine. Absorption of [3H]-L-Arg from the perfused loop, via an L-Arg- or L-homoarginine inhibitable process, was enhanced during HS. Salt restriction thus diminishes TGF attenuation by NO in the MD despite enhanced bNOS expression because of limited delivery and/or uptake of L-Arg via system y+. This defines a novel mechanism of renal microcirculatory adaptation to salt restriction via L-Arg-dependent changes in TGF. PMID- 9736255 TI - Ionic transport in macula densa cells. AB - Recent work has provided substantial insights into functional characteristics of macula densa (MD) cells. Microelectrode and patch-clamp experiments on the rabbit isolated thick ascending limb (TAL)/glomerulus preparation have shown that MD cells possess a furosemide-sensitive Na:K:2Cl cotransporter, an apical 41-pS K+ channel, and a dominant basolateral Cl- conductance. Increasing luminal fluid [NaCl] ([NaCl]L) results in furosemide-sensitive cell depolarization due to a rise in intracellular [Cl-] that stimulates basolateral electrogenic Cl- efflux. Intracellular pH (pHi) measurements show the presence of an apical Na:H exchanger that couples transepithelial Na+ transport to pHi. Experimental results and thermodynamic considerations allow estimation of intracellular [Na+] and [Cl-] ([Na+]i, [Cl-]i) under different conditions. When the Na:K:2Cl cotransporter is equilibrated (or in the presence of furosemide), [Na+]i and [Cl-]i are low (approximately 6 to 7 mM), whereas when the cotransporter is fully activated, [Na+]i and [Cl-]i increase substantially to approximately 70 and 20 mM, respectively. Finally, luminal addition of NH4+ produces cell acidification that depends on NH4+ apical transport rate through the Na:K:2Cl. Using a simple transport model for NH4+, the initial NH4+ influx rate in MD cells is comparable to the corresponding flux in TAL. This challenges the idea that MD cells have a low transport activity but supports our findings about large changes in intracellular concentrations as a function of [NaCl]L. PMID- 9736256 TI - Resetting protects efficiency of tubuloglomerular feedback. AB - Tubuloglomerular feedback (TGF) may effect long-term protection of total body salt and water or may govern minute-to-minute autoregulation of renal function. The task for which TGF is best suited depends on the orientation of ambient tubular flow relative to the inflection point of the TGF curve and on the tendency of TGF to reset in response to prolonged stimulation. Current data suggest that the TGF curve is coupled closely to ambient flow in individual nephrons such that the system is capable of compensating both negative and positive perturbations in tubular flow. This coupling is mediated by events within the juxtaglomerular apparatus that cause the TGF curve to reset laterally in response to sustained shifts in tubular flow. This resetting of TGF occurs within 30 to 60 minutes of an applied stimulus, suggesting that TGF is better suited to mediate dynamic autoregulation than to account for sustained vasoconstriction during proximal tubular injury. PMID- 9736257 TI - Tubuloglomerular feedback: its physiological and pathophysiological significance. AB - The mammalian nephron has a unique structure called juxtaglomerular apparatus (JGA); the primary function of the JGA includes tubuloglomerular feedback. Why is such a structure necessary? Analyses of available data strongly suggest that JGA has evolved to provide a fine tuning of the autoregulation of glomerular hemodynamics and high glomerular filtration rate in the face of very limited salt intake of our terrestrial environment, a function essential to allow a wide range of fluid and electrolyte intake with stable milieu interieur. Salt intake in excess is unique only to recent human cultures: salt intake is ordinarily less than 1 to 2 g per 60 kg of body weight in wildlife, including paleolithic humans. Any mutation or alteration of JGA function leading to renal salt conservation or maladaptive to high salt intake will not manifest in a low salt intake and thus would have been beneficial or inconsequential for survival in a natural environment, respectively. Thus, the mutation or alteration will be carried to subsequent generations. However, such altered function will result in essential hypertension or a maladaptation of JGA to high salt intake, which is a unique behavior of human civilizations of recent centuries. The kidney has not adapted to high salt intake through our evolution. PMID- 9736258 TI - Role of angiotensin in the congenital anomalies of the kidney and urinary tract in the mouse and the human. AB - The role of angiotensin in fluid and electrolyte and blood pressure homeostasis is well known. Recent developments indicate that angiotensin has a profound role not only in the developing urinary tract but also in the response of the urinary tract to specific noxious stimuli. Furthermore, the role of angiotensin II and its receptor has been understood quite poorly with respect to the developing renal unit. Knockout mice for the ATR2 gene show a significant incidence of congenital urinary tract anomalies. The congenital anomalies of the kidney and urinary tract (CAKUT) seen in these mice are very similar to the anomalies observed in humans. This has been supported further by the finding of an abnormality in the genetic sequence in patients with CAKUT. This article reviews experimental laboratory data as well as the potential implications for humans. PMID- 9736259 TI - Control of the renal renin system by local factors. AB - Local factors, such as prostaglandins (PGs), nitric oxide (NO), and endothelins (ETs), produced in the immediate vicinity of juxtaglomerular (JG) cells can exert significant effects on renin secretion and renin gene expression. PGE2, as the main renotubular PG, and PGI2, as the main endothelial prostanoid, both stimulate renin secretion and renin gene expression by activating cAMP formation in JG cells. Although the direct effect of NO on JG cells is less clear, its overall effect in vivo seems to be to stimulate the renin system. Evidence is emerging that stimulation by NO is related to the cAMP pathway, and cGMP-induced inhibition of cAMP-phosphodiesterase III (PDE-III) may mediate this effect. ETs, on the other hand, appear to inhibit the renin system, in particular in those pathways activated by cAMP, acting via Ca2+- and protein kinase C-related mechanisms. There is increasing evidence that both NO and PGs could be involved in the physiological regulatory mechanisms by which salt intake affects the renin system. PMID- 9736260 TI - A genetic approach for studying the role of thromboxane A2 in the kidney. AB - Thromboxane A2 (TxA2) has been implicated in a number of processes in normal kidney physiology and as a mechanism for injury in renal disease. TxA2 is a biologically active derivative of arachidonic acid and has potent vasoconstrictive and platelet-activation functions. Its actions are mediated by binding to specific G-protein-associated receptors designated TP receptors. There are at least two isoforms of the human TP receptor, and pharmacological evidence suggests TP receptor heterogeneity in other species. TP receptors are located in the renal cortex, and there may also be TxA2 binding sites in the medulla. TP receptors are involved in some normal functions of the kidney, and there is considerable evidence that TP receptors may mediate renal damage in disease states. To assess directly the role of the TxA2 in normal kidney function and in murine models of human disease, we have used gene targeting to eliminate expression of the TP receptor in mice. PMID- 9736261 TI - Regulation of renal function by prostaglandin E receptors. AB - Prostaglandin E2 is the major cyclooxygenase product of arachidonic acid metabolism produced along the nephron. This autacoid interacts with four distinct, G-protein-coupled E-prostanoid receptors designated EP1-EP4. The intrarenal distribution of each receptor has been mapped and the consequences of receptor activation examined. EP3 receptor mRNA is expressed highly in the medullary thick ascending limb (mTAL) and collecting duct (CD). EP3 receptor activation inhibits cAMP generation via Gi, thus inhibiting vasopressin stimulated water reabsorption in the CD. EP3 receptor activation also may contribute to PGE2-mediated inhibition of NaCl absorption in the mTAL. The EP1 receptor is coupled to increased cell [Ca2+]. EP1 mRNA expression is restricted to the CD, and receptor activation inhibits Na+ absorption. PGE2 also increases cAMP generation in the cortical thick ascending limb and CD; this may be due to EP4 receptor activation. EP4 mRNA is readily detected in the CD with little detectable EP2 expression. The EP4 receptor appears to be expressed both on luminal and basolateral membranes. EP4 receptor activation also may contribute to the regulation of renin release by the juxtaglomerular apparatus. The consequences of renal EP-receptor activation for salt and water balance may be determined by the relative renal expression of each of these receptors. PMID- 9736262 TI - Physiological and pharmacological implications of AT1 versus AT2 receptors. AB - Angiotensin II (Ang II) has diverse physiological actions that lead, for instance, to increases in extracellular volume and peripheral vascular resistance and blood pressure, and it has also been implicated in the regulation of cell growth and differentiation. Molecular cloning and pharmacological studies have defined two major classes of Ang II receptors, designated AT1 and AT2. Most effects of Ang II are mediated by AT1 receptors. Much less is known about the physiological role of AT2 receptors. Recent evidence suggests involvement of AT2 receptors in development, cell differentiation, apoptosis, and regeneration in various tissues. AT1 and AT2 receptors have been shown to exert counteracting effects on cellular growth and differentiation, vascular tone, and the release of arginine vasopressin. In each condition, the AT2 receptor appears to down modulate actions mediated by the AT1 receptor, resulting in decreased cellular proliferation, decreased levels of serum arginine vasopressin levels, or decreased vasoconstrictor responses. In addition, in neuronal cell lines, the AT2 receptor exerts antiproliferative actions and promotes neurite outgrowth, an effect accompanied by significant changes in the expression pattern of growth/differentiation-related genes. PMID- 9736263 TI - Shear stress and the endothelium. AB - Shear stress and the endothelium. Vascular endothelial cells (ECs) in vivo are influenced by two distinct hemodynamic forces: cyclical strain due to vessel wall distention by transmural pressure, and shear stress, the frictional force generated by blood flow. Shear stress acts at the apical cell surface to deform cells in the direction of blood flow; wall distention tends to deform cells in all directions. The shear stress response differs, at least partly, from the cyclical strain response, suggesting that cytoskeletal strain alone cannot explain it. Acute shear stress in vitro elicits rapid cytoskeletal remodeling and activates signaling cascades in ECs, with the consequent acute release of nitric oxide and prostacyclin; activation of transcription factors nuclear factor (NF)kappaB, c-fos, c-jun and SP-1; and transcriptional activation of genes, including ICAM-1, MCP-1, tissue factor, platelet-derived growth factor-B (PDGF B), transforming growth factor (TGF)-beta1, cyclooxygenase-II, and endothelial nitric oxide synthase (eNOS). This response thus shares similarities with EC responses to inflammatory cytokines. In contrast, ECs adapt to chronic shear stress by structural remodeling and flattening to minimize shear stress. Such cells become very adherent to their substratum and show evidence of differentiation. Increased adhesion following chronic shear stress has been exploited to generate vascular grafts with confluent EC monolayers, retained after implantation in vivo, thus overcoming a major obstacle to endothelialization of vascular prostheses. PMID- 9736264 TI - Epithelial sodium channel regulatory proteins identified by functional expression cloning. AB - We describe here our current strategy for identifying and cloning proteins involved in the regulation of the epithelial sodium channel (ENaC). We have set up a complementation functional assay in the Xenopus laevis oocyte expression system. Using this assay, we have been able to identify a channel-activating protease (CAP-1) that can increase ENaC activity threefold. We propose a novel extracellular signal transduction pathway controlling ionic channels of the ENaC gene family that include genes involved in mechanotransduction (degenerins), in peptide-gated channels involved in neurotransmission (FaNaCh), in proton-gated channels involved in pH sensing (ASIC) or pain sensation (DRASIC). PMID- 9736265 TI - A role for endothelin in the pathogenesis of hypertension: fact or fiction? AB - Endothelin-1 (ET-1) was discovered 10 years ago. Because it is one of the most potent vasoconstrictors in vivo, a pathophysiological role for the peptide as a mediator of hypertension has been postulated. Several clinical studies, however, have been unable to identify elevated ET levels in the plasma of hypertensive patients, suggesting that it does not play a prominent role in this disease. More recently, evidence has been presented that ETs act predominantly at the autocrine/paracrine level and that measurements of plasma levels can give only an indirect view of the activity of the system. In addition, transgenic technology has uncovered new actions of the peptide systems in recent years, which point to a key function of the system in prenatal development. Moreover, investigation of conditions associated with hypertensive end-organ damage, such as chronic renal failure, has led to a re-evaluation of the role of the ET system in hypertension. This article discusses this recent evidence and defines the exact role of the ET system in hypertension and hypertensive end-organ damage. PMID- 9736266 TI - Myogenic effects enhance norepinephrine constriction: inhibition by nitric oxide and felodipine. AB - Myogenic, pressure-induced vasoconstriction may amplify the effects of circulating vasoconstrictors. Through intravital microscopy in cremaster arterioles (31 to 115 microm diameter), the relative contribution of myogenic responses (MR) to norepinephrine (NE)-induced constriction and the inhibitor potency of nitric oxide (NO) or a Ca2+ entry blocker (CEB), felodipine (F), were examined. In 24 anesthetized hamsters, a vessel occluder was placed around the aorta to control cremaster vessel inflow pressure (IP). NE infusion increased blood pressure (by 50 +/- 2 mm Hg) and induced significant constriction (24% +/- 9%) in small arterioles (< 65 microm) only. The constriction, which was not altered by adrenergic blockade, was dependent on the actual IP and was abolished when the IP increase was blocked. NO synthase (NOS) blockade unmasked a significant MR in large arterioles. F inhibited the MR predominantly in large vessels. In isolated microvessels, F completely blocked the pressure-induced Ca2+ increase and MR. We conclude that circulating NE constricts muscle arterioles mainly by a myogenic mechanism. NO effectively opposes MR in larger arterioles, thus restricting MR and vasoconstrictor reinforcement to a small section of the vasculature being tightly controlled by metabolic signals. MR, which otherwise would impair adjustment of peripheral resistance, is reduced by CEB predominantly in larger arterioles, similar to NO. PMID- 9736267 TI - Pathophysiology of renal fluid retention. AB - Central to a unifying hypothesis of body fluid regulation is maintenance of arterial circulatory integrity. This may be disturbed by arterial underfilling, either from reduction in cardiac output or by peripheral arterial vasodilation. In cardiac failure (CF), cardiac output falls and the nonosmotic release of arginine vasopressin (AVP) and expression of AVP mRNA in the hypothalamus are stimulated. V2 AVP receptor antagonists correct the impaired water excretion in rats with low-output CF, increase solute free water clearance, correct the hyponatremia in congestive CF patients, and normalize urinary concentrations of the aquaporin-2 (AQP-2) water channels. In conditions associated with peripheral vasodilation, such as cirrhosis, nonosmotic release of AVP also occurs, and AQP-2 gene expression in the rat kidney is up-regulated. In cirrhosis, nitric oxide mediated vasodilation occurs early prior to water retention. V2 antagonists reverse the latter. In normal pregnancy, plasma AVP is relatively high for the degree of hypoosmolality. Pregnant rats up-regulate AQP-2 in the renal papilla, an effect reversed by V2 receptor antagonists. This supports the hypothesis that AVP is an important mediator of renal water retention in pregnancy. In summary, AVP-mediated water retention through collecting duct AQP-2 water channels is important in both low-output CF and high-output states such as cirrhosis and pregnancy. V2 receptor antagonists reverse the water retention and down-regulate AQP-2 water channels. PMID- 9736268 TI - Adaptation of Madin-Darby canine kidney cells to hypertonic medium: an electron microprobe analysis. AB - Madin-Darby canine kidney (MDCK) cells of confluent epithelial sheets grown on permeable supports respond to hyperosmotic stress by short- and long-term regulatory volume increase (RVI). Although short-term RVI includes the uptake of inorganic electrolytes, long-term RVI does not and seems therefore to result from accumulation of organic osmolytes. PMID- 9736269 TI - Cationic amino acid transporter mRNA expression in rat kidney and liver. AB - Expression of rat cationic amino acid transporter 2 (r-CAT-2) mRNA was studied in kidney and liver using Northern blot analysis and nonradioactive in situ hybridization with a probe identifying both the r-CAT-2alpha and -2beta splice variants. Expression of r-CAT-2 mRNA was higher in the liver than in the kidney. Within the kidney, r-CAT-2 mRNA was more abundant in the outer and inner medulla than in the cortex. In the liver lobule, the intensity of the hybridization signal in hepatocytes decreased between the portal area and the central vein. In the kidney, hybridization signals were detected in parietal cells of Bowman's capsule, various tubule cells of outer and inner medulla, in endothelial and interstitial cells of inner medulla, and in papillary epithelial cells. PMID- 9736270 TI - Video-imaging microfluorometry identifies alpha- and beta-like cell types in Madin-Darby canine kidney monolayers. AB - On the basis of intracellular, accumulation of c-SNAFL-2, we have identified three cell subtypes in Madin-Darby canine kidney (MDCK) monolayers. Highly fluorescent cells (HFC) have a high intracellular pH (pHi, whereas cells with medium fluorescence (MFC) have low pHi when perfused with buffer containing 125 mM Cl-. HFC express a Cl-/HCO3- exchanger on the apical but not the basolateral membrane. MFC express a Cl-/HCO3- exchanger on the basolateral but not the apical membrane. We have termed these cells beta- and alpha-MDCK cells, respectively. Cells with low fluorescence (LFC) probably extrude c-SNAFL-2 through a monocarboxylate transporter, because p-4-(chloromercuri)phenylsulfonic acid (PCMBS), an inhibitor of this transporter, leads to homogeneous fluorescence. PMID- 9736271 TI - Characteristics of sodium uptake across the basolateral membrane of oxyntic cells. AB - To characterize further serosal Na uptake into gastric oxyntic cells under resting conditions, cellular element concentrations were determined in isolated frog (Rana temporaria) gastric mucosae using electron microprobe analysis. The epithelia were kept short circuited in Ussing-type chambers, and element analysis was performed on freeze-dried cryosections. After ouabain (10(-4) M), the [Na] in oxyntic cells increased within 30 to 60 minutes from approximately 25 to 100 mmol/kg wet wt, and [K] decreased similarly (from 100 to 25 mmol/kg wet wt). These changes occurred regardless of whether the basolateral incubation medium contained HCO3 or N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES) as buffers. When, prior to the addition of ouabain, 10(-3) M amiloride was applied to the serosal side to inhibit the Na-H antiporter, the ouabain-induced increase in cellular [Na] was prevented completely in HEPES-, but not in HCO3-Ringer. The data are compatible with the notion that Na is taken up by a Na-H antiporter and a Na-HCO3 symporter. At least under these experimental conditions, these transporters seem to contribute substantially to basolateral Na uptake in oxyntic cells. PMID- 9736272 TI - Angiotensin-dependent gene expression in the developing rat kidney. AB - We aimed to identify genes involved in the growth effects of angiotensin II (Ang II) during kidney development. In rats treated from birth with the Ang II type-1 receptor blocker losartan, expression of transforming growth factor beta1 (TGF beta1), platelet-derived growth factor B (PDGF-B), vascular endothelial growth factor (VEGF), and epidermal growth factor (EGF), as measured by Northern blot, did not change significantly (N = 4 to 6 per group each). Differential display methods, used to identify genes with Ang II-dependent expression, produced mostly false positives. We identified one novel rat partial cDNA, termed AD.5, that is related to a human orphan receptor. AD.5 was expressed in a developmentally regulated pattern and may be involved in kidney development and/or the trophic actions of Ang II. PMID- 9736273 TI - Chloride channels ClC-2 and ICln mRNA expression differs in renal epithelial ontogeny. AB - Development-dependent mRNA expression of the chloride channels ClC-2 and ICln was studied by quantitative reverse transcriptase-polymerase chain reaction in rat ureteric bud and cortical collecting duct primary monolayer cultures. Abundance of ClC-2 mRNA increased in ureteric bud cells between embryonic day 15 (E15) and E17, peaked at postnatal day 3 (P3), and was down-regulated at P7 when morphogenesis is complete, suggesting a specific embryonic function. Expression of ICln mRNA, in contrast, up-regulated continuously with development. PMID- 9736274 TI - Pleiotropic effects of hepatocyte growth factor in proximal tubule involve different signaling pathways. AB - Hepatocyte growth factor (HGF) accelerates renal tubule cell regeneration and induces tubulogenic differentiation via the intracellular tyrosine kinase (TK) domain of its receptor, the proto-oncogene c-Met. We tested whether different signaling pathways may be involved by examining HGF binding and effects on cell proliferation, migration, scattering, and tubulogenic differentiation in the bipolar differentiating rabbit proximal tubule cell line PT-1 under serum-free conditions in the presence or absence of the protein TK inhibitors (PTKIs) herbimycin-A, genistein, methyl-2,5-dihydroxycinnamate, and geldanamycin. These PTKIs inhibit pp60(c-src), a nonreceptor TK involved in cell-growth control. HGF bound to a single high-affinity receptor class, increased microvilli numbers 1.5 fold, enhanced cell proliferation and migration 1.8-fold, and stimulated formation of tubule structures 2.2-fold. PTKI inhibited the mitogenic and motogenic effects of HGF with different potencies and comparable maximal effects but had no specific influence on HGF-induced tubulogenic cell differentiation. These data underline the importance of pp60(c-src) in mediating mitogenic and motogenic effects of HGF, whereas stimulation of tubulogenic cell differentiation may be transduced by a pp60(c-src)-independent pathway. PMID- 9736275 TI - Effects of hypoxia on renin secretion and renal renin gene expression. AB - Plasma renin activity (PRA) and renal renin mRNA levels were measured in male rats exposed to hypoxia (8% O2) or to carbon monoxide (CO; 0.1%) for six hours. PRA increased fourfold and 3.3-fold, and renin mRNA levels increased to 220% and 200% of control, respectively. In primary cultures of renal juxtaglomerular (JG) cells, hypoxia (lowering medium O2 from 20% to 3% or 1%) for 6 or 20 hours did not affect renin secretion or gene expression. Renal denervation did not prevent stimulation of the renin system by hypoxia. Because norepinephrine increased 1.7 fold and 3.2-fold and plasma epinephrine increased 3.9-fold and 7.8-fold during hypoxia and CO inhalation, respectively, circulating catecholamines might mediate the stimulatory effects of hypoxia on renin secretion and renin gene expression. Stimulation of beta-adrenergic receptors by continuous infusion of 160 microg/kg/hr isoproterenol increased PRA 17-fold and 20-fold after three and six hours, respectively, and renin mRNA by 130% after six hours. In rats with a stimulated renin system (low-sodium diet), isoproterenol did not stimulate PRA or renal renin mRNA further. In summary, both arterial and venous hypoxia can stimulate renin secretion and renin gene expression powerfully in vivo but not in vitro. These effects seem not to be mediated by renal nerves or by a direct effect on JG cells but might be mediated by circulating catecholamines. PMID- 9736276 TI - Detection of multiple vascular endothelial growth factor splice isoforms in single glomerular podocytes. AB - Glomerular podocytes are major determinants of filtration permselectivity in the glomerulus. Although the molecular mechanisms determining the characteristics of the glomerular filtration unit are incompletely understood, vascular endothelial growth factor (VEGF) has been implicated. To analyze this process in situ, we established a method that allows exploration of in vivo mRNA expression of podocytes using single-cell reverse transcriptase-polymerase chain reaction (RT PCR). Microdissected mouse glomeruli were held in a patch-clamp apparatus, and single podocytes were harvested by aspiration. After lysis, the cells were reverse transcribed, and PCR was performed (45 cycles). The podocyte nature of the material was confirmed by detection of podocyte-specific mRNA (glomerular epithelial protein 1 and Wilms' tumor protein 1). Using specific oligonucleotide primers, VEGF was detected in mRNA obtained from renal cortex, single microdissected glomeruli, cultured murine podocytes, and single podocytes in situ. All cells examined expressed three VEGF isoforms (121, 165, and 189). These differ in their capacity for binding to extracellular matrix and could have different potencies regulating glomerular endothelial permeability. Our approach should allow a semiquantitative, isoform-specific evaluation of VEGF mRNA expression in podocytes during nephrogenesis and in glomerular disease. PMID- 9736277 TI - Influence of osmotic stress on heat shock proteins 25 and 72 in mouse mesangial cells. AB - Previous studies have shown intense staining for heat shock protein 25 (HSP25) in the extraglomerular mesangium (EGM). Because relationships are believed to exist between osmotic stress, expression of HSP25, and protection against stress and because the EGM may be exposed to high local tonicity, we examined the expression of HSP25 and the major stress-inducible and cytoprotective HSP72 in mouse mesangial cells and embryonic lung fibroblasts (3T3) after exposure to hypertonic stress (addition of 150 mM NaCl to the medium for two to seven days). Mesangial, but not 3T3, cells expressed high levels of HSP25 already under control conditions, whereas neither cell line contained HSP72. Hypertonic treatment neither enhanced (mesangial cells) or induced (3T3 cells) HSP25 expression. HSP72, however, was induced strongly in 3T3 cells, but only minimally in mesangial cells. The high level of HSP25 in mesangial cells thus seems not to be a consequence of high tonicity in the EGM because cultured mesangial cells express HSP25 already under control conditions, and osmotic stress did not induce HSP25 in either cell line. Furthermore, high amounts of HSP25 seem to reduce the requirement for HSP72 after stress exposure, suggesting that, in mesangial cells, HSP25 might assume some functions of HSP72. PMID- 9736278 TI - Hypertonicity affects heat shock protein 27 and F-actin localization in Madin Darby canine kidney cells. AB - High concentrations of NaCl are known to perturb the cytoskeleton. In this study, expression and intracellular localization of actin, an important component of the cytoskeleton and of heat shock protein (HSP)27, which promotes the assembly of F actin, were examined in Madin-Darby canine kidney (MDCK) cells grown chronically in hypertonic medium. HSP27 mRNA abundance was increased twofold compared with wild-type MDCK cells. Chronic hypertonic stress led to enrichment of HSP27 in the insoluble component of the cell lysate and colocalization with cortical F-actin. These results support the notion that HSP27 participates in the modulation of actin dynamics following hypertonic stress. PMID- 9736279 TI - Angiotensin II modulates cellular functions of podocytes. AB - Angiotensin II modulates cellular functions of podocytes. The aim of this study was to examine the effects of angiotensin II (Ang II) on membrane voltage (Vm) and cytosolic calcium activity ([Ca2+]i) of rat podocytes. To approach better the in vivo situation, we have developed an experimental approach that allows podocytes to be studied in the intact microdissected glomerulus. Ang II depolarized podocytes in the glomerulus (EC50 15 nM, N = 49). Like podocytes in the glomerulus, podocytes in short-term culture also depolarized in response to Ang II (10 nM, N = 5). Ang II increased [Ca2+]i in podocytes in culture (EC50 3 nM, N = 229). In a solution with reduced extracellular [Ca2+] (10 microM), Ang II mediated [Ca2+]i increase was significantly reduced by 60% +/- 20% (N = 12). Flufenamate, an inhibitor of nonselective ion channels, inhibited Ang II-mediated increase of [Ca2+]i (IC50 20 microM, N = 29). The Ang subtype 1 (AT1) receptor antagonist losartan inhibited both Ang II-mediated depolarization and [Ca2+]i increase in podocytes (N = 5 to 35). Our results support the concept that Ang II might influence podocyte function directly via an AT1 receptor. PMID- 9736280 TI - Reverse transcriptase-polymerase chain reaction study of anion exchanger-2 in canine tissues and different Madin-Darby canine kidney cell types. AB - Reverse transcriptase-polymerase chain reaction (RT-PCR) of mRNA from canine large intestine, skeletal muscle, pancreas, kidney, and spleen and from cultured wild-type and C7 and C11 Madin-Darby canine kidney (MDCK) cells revealed considerable variation in anion exchanger (AE)1 and AE2 mRNA levels between the tissues. Similar high levels of AE2 mRNA were detected in all the MDCK cell populations. AE2 in MDCK cells is probably the basolateral Cl-/HCO3- exchanger common to the principal and beta-intercalated cells. PMID- 9736281 TI - Effect of ischemia on localization of heat shock protein 25 in kidney. AB - The effects of renal ischemia on the intracellular distribution of the low molecular weight heat shock protein (HSP)25 were examined using immunofluorescence microscopy. In all kidney zones, ischemia decreased HSP25 in the supernatant of the tissue homogenates and increased it in the pellet fraction (containing mainly nuclei and cytoskeletal components). This was associated with disappearance of HSP25 staining from the brush border of proximal convoluted tubule (PCT) cells. Because no nuclear staining of cortical tubule cells was apparent either in control or ischemic kidneys, ischemia seems to cause a closer association of HSP25 with cytoskeletal components. HSP25 probably participates in the postischemic restructuring of the cytoskeleton of PCT cells. PMID- 9736283 TI - Influence of extracellular pH on intracellular pH in arterioles and skeletal muscle of spontaneously hypertensive and Wistar-Kyoto rats. AB - The effect of changing extracellular pH (pHo) on intracellular pH (pHi) in mesenteric arterioles of spontaneously hypertensive (SHR) and Wistar-Kyoto (WKY) rats was investigated using the fluorescent indicator seminaphthorhodafluor c SNARF-1 to measure pHi. Although the pHi of arterioles from WKY varied directly with pHo, that of SHR arterioles varied inversely. This abnormal pHi regulation in SHR was corrected by 50-microM 5-(N-ethylisopropyl)amiloride (EIPA). Vmax of H+ transport in response to changing extracellular Na+ was higher in SHR than in WKY, but Km did not differ. The Hill coefficient for H+ transport with respect to pH; was 1.69 in prehypertensive and 1.56 in hypertensive SHR. These results indicate that the Na+/H+ exchanger is particularly hyperactive in mesenteric arterioles of SHR. PMID- 9736282 TI - Luminal signal in tubuloglomerular feedback: what about potassium? AB - Evidence suggests that a minimal luminal [K+] is required to elicit a full tubuloglomerular feedback (TGF) response, consistent with transmission of the TGF signal across the macula densa (MD) via the Na+-2Cl(-)-K+ cotransporter. Furthermore, it appears that luminal [K+] at the MD is close to the K+ affinity of the Na+-2Cl(-)-K+ cotransporter and changes in response to altering late proximal tubular flow rate (VLP), that is, a maneuver that induces a TGF response. These findings suggest that luminal [K+] (besides [Cl-]) could be rate limiting in TGF. In the thick ascending limb of Henle's loop (TALH), most of the luminal K+ is derived from recycling across the apical tubular membrane. Because changing VLP causes relatively greater alterations in the absolute Na+ and Cl- delivery to Henle's loop than in K+ load, the parallel changes of VLP and luminal [K+] at the MD, despite significant alteration in K+-dependent reabsorption of Na+ and Cl- via the Na+-2Cl(-)-K+ cotransporter, imply a transport-dependent adaptation of K+ recycling in TALH. PMID- 9736284 TI - Expression of the Na+/myo-inositol cotransporter in the juxtaglomerular region. AB - Myo-inositol is a major compatible osmolyte in the renal medulla and is accumulated in cells under hypertonic conditions by uptake via a Na+/myo-inositol cotransporter (SMIT). SMIT is regulated by extracellular osmolarity at the transcription level. We investigated localization of SMIT in rat kidney by immunohistochemical staining using an anti-SMIT-antibody raised against a synthetic peptide corresponding to part of SMIT and by in situ hybridization. SMIT protein localized predominantly to the basolateral membranes of cells of the thick ascending limb of Henle (TAL) and inner medullary collecting duct (IMCD). Macula densa (MD) cells, identified as the Tamm-Horsfall-protein (THP)-unreactive cells surrounded by THP-reactive TAL cells, also stained for anti-SMIT. In situ hybridization yielded the intense SMIT signals in the TAL and IMCD and also in the juxtaglomerular (JG) region. Prior loading of the animal with a high concentration of NaCl rapidly induced SMIT mRNA; furosemide down-regulated it. The high level of SMIT expression suggests that MD cells are exposed to hypertonicity at the basolateral surface. Because SMIT expression seemed to be proportional to the magnitude of NaCl reabsorption, it may be a good marker for examination of the tubuloglomerular feedback mechanism in vivo. PMID- 9736285 TI - Human glomerular structure under normal conditions and in isolated glomerular disease. AB - Investigation of the human glomerulus in health and disease shows that the human glomerulus comprises seven lobule-like structures with numerous anastomoses. The total length of the capillaries in a single glomerulus is 0.95 cm, making a total of 19 km for all 2-million glomeruli. The total surface area of all glomerular capillaries is 6,000 cm2. The total filtration surface area is 516.1 cm2. Severe isolated disease of the glomerulus, as seen in acute endocapillary glomerulonephritis, membranoproliferative glomerulonephritis types I and II, membranoproliferative glomerulonephritis plus chronic membranous glomerulonephritis, diabetic glomerulosclerosis, and glomerular amyloidosis, does not lead to elevation of serum creatinine concentration, even if the filtration area is reduced to about 20% (as in diabetes) of the normal value. It is concluded that isolated glomerular disease does not lead to elevation of the serum creatinine concentration. Glomerulopathies in which there is acute or chronic elevation of the serum creatinine concentration are accompanied by acute renal failure or involvement of the renal cortical interstitium, respectively. PMID- 9736287 TI - Adenosine and extracellular volume in radiocontrast media-induced nephropathy. AB - Renal hemodynamic changes could play a key role in radiocontrast media-induced nephropathy (RCIN), although the pathophysiological mechanisms are unclear. We investigated the role of adenosine in RCIN caused by sodium diatrizoate (Urografin, 3 ml/kg) in nitro-L-Arg methyl ester (L-NAME)-hypertensive rats in different hydration states [eight weeks of L-NAME (50 mg/liter) in drinking water; high or low sodium intake for the last two weeks]. In clearance experiments under thiobutabarbital anesthesia in these previously mentioned animals, glomerular filtration rate (GFR), renal blood flow (RBF), and mean arterial pressure (MAP) were measured in the presence or absence of the adenosine A1-receptor antagonist 8-cyclopropyl-1,3-dipropylxanthine (DPCPX, 100 microg/kg bolus plus 10 microg/kg/hr). DPCPX or pretreatment did not change control hemodynamics. Contrast medium caused GFR and RBF to fall significantly in volume depleted rats (from 0.29 +/- 0.02 to 0.21 +/- 0.02 ml/min/100 g and 5.4 +/- 0.3 to 4.0 +/- 0.4 ml/min, respectively) without change in MAP. In volume-expanded rats, changes were not significant (0.25 +/- 0.01 to 0.24 +/- 0.02 ml/min/100 g and 5.6 +/- 0.3 to 5.3 +/- 0.4 ml/min, respectively). In the volume-depleted rats, changes were prevented by DPCPX (0.27 +/- 0.02 to 0.24 +/- 0.02 ml/min/100 g and 4.8 +/- 0.1 to 5.0 +/- 0.1 ml/min, respectively). The acute hemodynamic effects elicited by contrast medium in L-NAME hypertensive rats thus can be prevented by volume expansion. Adenosine, via A1-receptors, contributes to the adverse effects of contrast media. PMID- 9736286 TI - Different activation of mitogen-activated protein kinases in experimental proliferative glomerulonephritis. AB - Mitogen-activated protein (MAP) kinases are critical for cell signaling goals such as cellular proliferation and induction of apoptosis. We examined whether MAP kinases, as a point of convergence for multiple extracellular stimuli, are activated in proliferative glomerulonephritis (GN) in vivo. Accelerated crescentic anti-glomerular basement membrane (GBM) GN was induced in rats preimmunized with rabbit IgG by administration of rabbit anti-rat GBM serum. Whole cortical tissue and isolated glomeruli were then subjected to kinase activity assays and Western blot analysis. Cortical activity of the archetypal MAP kinase, extracellular signal-regulated kinase (ERK), was increased significantly one, three, and seven days after induction of GN. In contrast, activation of MAP kinases with antiproliferative actions, stress-activated protein kinase, and p38 MAP kinase was detectable only in the early stages of proliferative GN (days one and three), implying that different MAP kinases serve distinct roles in the pathogenesis of GN. PMID- 9736288 TI - Insulin-like growth factor-I restores microvascular autoregulation in experimental chronic renal failure. AB - Impairment of autoregulation (AR) is associated with accelerated progression of chronic renal failure (CRF). As the bioavailability of insulin-like growth factor I (IGF-I) is low in CRF, we investigated the effects of acute luminal application of 10 nM recombinant human IGF-I on AR in juxtamedullary (JM) afferent arterioles (AA) perfused in vitro with a blood solution [(approximately 30% hematocrit (HCT)]. Studies were conducted in AA from adult male rats three to four weeks after five-sixths nephrectomy (Nx) by either surgical excision (N = 7) or infarction (N = 5) of two thirds of the remnant kidney; controls (N = 6) had sham surgery. AA from both Nx groups exhibited marked hypertrophy and impaired AR responses (60 to 140 mm Hg perfusion pressure), features more pronounced in the infarction group. Responses to abluminal acetylcholine (10 microM) were similar in sham and excision groups but were significantly blunted in the infarction group. All groups vasodilated significantly after Ca-channel blockade (10 mM MnCl2). IGF-I restored AR in AA from both Nx groups (P < 0.05, analysis of variance) while it vasodilated AA from controls. These results suggest that IGF-I may protect the glomerulus from injury by maintaining autoregulatory control of renal blood flow, thereby slowing the progression of CRF. PMID- 9736289 TI - Endothelial vasoconstrictor prostanoids, vascular reactivity, and acute renal failure. AB - The interaction of endothelium-derived vasoconstrictor prostaglandins, the angiotensins (Ang), and the sympathetic nervous system in acute renal failure still remains to be determined. In this study, acute renal failure (ARF) was induced in male Wistar Kyoto rats (N = 7) in a 2K/2C model of 30-minute clamping. Contractions to Ang I and II and norepinephrine (NE) were studied in isolated aortic and renal artery rings 24 hours after clamp release. Sham-operated animals served as controls (N = 7). In ARF, contractions to NE were increased in the aorta and even further enhanced in the renal artery (P < 0.05 to 0.001), whereas contractions to Ang I and II were blunted (P < 0.05). Contractions were inhibited by SQ 30741, a thromboxane A2 (TXA2)/prostaglandin H2 (PGH2) receptor antagonist. We conclude that ARF is characterized by abnormal vascular reactivity both in the renal as well as the systemic vasculature that is in part mediated by endothelium derived vasoconstrictor prostaglandins. PMID- 9736290 TI - Preventive strategies in endothelin-induced renal failure. AB - The endothelial vasoconstrictor endothelin (ET) can induce acute renal failure when fibrinolysis and vasodilatory prostanoids (PGs) are inhibited. This study compares therapeutic agents preventing ET-induced acute renal failure in anesthetized female pigs. We investigated the effect of four ET boli (1.5 microg/kg, i.v.) after pretreatment with indomethacin (2 mg/kg) and epsilon aminocaproicacid (100 + 50 mg/kg) alone (controls, group 1) or during additional nifedipine (10 microg/kg/h; group 2), hirudin (0.5 mg/kg; group 3), or enalapril (2 x 0.15 mg; group 4) on coagulation, PGs, and renal function. The ET-induced blood pressure increase was lower in groups 2 to 4 (lowest in group 3, P < 0.05). PG synthesis was blocked in all groups. The initial hypercoagulability (controls) resulted in disseminated intravascular coagulation that was prevented by hirudin and was attenuated in groups 2 and 4. At the end of the experiment, creatinine clearance was significantly (P < 0.05) decreased. The recovery of renal function two hours after the last ET bolus was most pronounced in the hirudin group. All therapeutic drugs attenuated ET-induced impairment of renal function. Hirudin seems to be the most potent protective drug. Prevention of further ET release evoked by ET-mediated secretion of thrombin might explain this. These results suggest three important pathways for ET's hemodynamic and renal effects. PMID- 9736291 TI - Renal morphology in essential hypertension: analysis of 1177 unselected cases. AB - Morphological and clinical analysis of 1177 renal biopsies from nonselected patients with essential hypertension revealed compensated benign nephrosclerosis in 775 cases. Decompensated benign nephrosclerosis was found in 251 cases, and secondary malignant nephrosclerosis was found in 151 cases. This article describes the morphological and clinical features of decompensated benign nephrosclerosis, which has received little recognition until now. The morphological and clinical features of secondary malignant nephrosclerosis, which is induced by hypertension, are also considered. There is also a discussion of the differentiation of the latter from primary malignant nephrosclerosis, in which stenosis of the preglomerular vessels develops in the presence of normal blood pressure and leads secondarily to renal hypertension. PMID- 9736292 TI - Renovascular parathyroid hormone-related protein in spontaneously hypertensive rats: dilator or trophic factor? AB - Parathyroid hormone-related protein (PTHrP) is expressed throughout the renovascular system, and it dilates renal vessels, increases renal blood flow and glomerular filtration rate, and stimulates renin release. Mechanical forces and experimental hypertension have been shown to stimulate PTHrP expression in smooth muscles, suggesting a negative feedback control of vascular tone by PTHrP in hypertension. In this study, we compared the impact of a PTHrP receptor antagonist, PTHrP (7-34), and a PTHrP receptor agonist, PTHrP (1-36), on the vascular resistance of perfused kidneys isolated from spontaneously hypertensive rats (SHR) and Wistar Kyoto rats (WKY). Endogenous PTHrP appears not to act as a renal vasodilator in either WKY or SHR. However, the vasodilation following infused PTHrP (1-36) is blunted markedly in SHR, possibly due to desensitization or down-regulation of PTH/PTHrP receptors. Negative feedback control of vascular tone by PTHrP in SHR thus appears unlikely. The results raise the question of whether endogenous renovascular PTHrP behaves rather as a growth factor than as a vasodilator. PMID- 9736293 TI - Glomerulus number and blood pressure in the Prague hypertensive rat. AB - The kidney has long been attributed a key role in the pathogenesis of hypertension. Reduction of filtration area by glomerular loss is regarded currently as a major causative mechanism. Here we analyze the relationship between glomerulus number and blood pressure (BP) in a new model of genetic hypertension and the Prague hypertensive rat (PHR) and the Prague normotensive rat (PNR). Glomerular numbers were determined in 7- to 53-week-old PNR and PHR, and the correlation with conscious systolic BP was analyzed. PHR had significantly higher BP but 19% fewer glomeruli than PNR. Glomerular number correlated (partial correlation analysis, controlling for effects of body weight, age, and kidney weight) significantly (P < 0.01, r2 = 0.46) with BP in male PHR but not in female PHR or in PNR. Moreover, subgroups of PHR and PNR selected for the same mean BP showed the same differences in glomerular counts, and subgroups selected for the same mean glomerular count showed the same differences in BP as the whole group. Reduced glomerular numbers and BP seem not to be causally related to BP in PHR older than seven weeks. Other mechanisms, such as genetically determined changes in transporter and receptor proteins, vascular abnormalities, and humoral mechanisms, must be considered. PMID- 9736294 TI - Sodium, angiotensin II, blood pressure, and cardiac hypertrophy. AB - Blood pressure (BP) in rats was elevated intermittently by i.p. injections of angiotensin II (Ang II; 200 microg/kg), and the effect on cardiac index was determined. The BP response was assessed in selected rats by telemetry. Elevation of BP between 8:00 and 12:00 produced cardiac enlargement similar to that produced by continuous Ang II infusion, and the response correlated better with the acute BP elevation than with 24-hour cardiac work. A high-sodium diet also increased left-ventricular hypertrophy (LVH) without a major effect on BP. The addition of Ang II intensified this response. A low-sodium diet had no significant effect on BP or on cardiac size, but prevented the cardiac hypertrophy produced by Ang II without altering the BP response. These results suggest that acute BP elevation, probably working through increased wall tension, is a more potent stimulus for cardiac hypertrophy than 24-hour workload. The sodium intake of the rat plays an important role influencing the cardiac but not the BP response to Ang II. These results infer that it is important to avoid episodes of acute BP elevation. PMID- 9736295 TI - Hypertension and impaired renal angiotensin II response in rats after chronic neuronal nitric oxide synthase inhibition. AB - Nitric oxide (NO) produced by the macula densa cells is important for the control of tubuloglomerular feedback (TGF). Reduced production of NO by these cells activates TGF and could result in hypertension, although the TGF activity is then normalized in the hypertensive state. The normalization of TGF in this form of hypertension might be explained by an impaired ability of angiotensin II to constrict renal vessels or by up-regulation of some other vasodilator not affected by NO synthase inhibitors. PMID- 9736296 TI - Role of nitric oxide and thromboxane in the maintenance of cerebrovascular tone. AB - This study investigated the role of thromboxane A2 (TXA2) and neuronal nitric oxide (NO) synthase (nNOS)-derived NO in the maintenance of resting cerebrovascular tone. Rat basilar artery (BA) segments were mounted in myographs to study their isometric tension development. 7-Nitro indazole monosodium salt (7 NINA), a specific inhibitor of nNOS, had no significant effect on the resting tone, whereas the general NOS blocker N(G)-nitro-L-arginine (L-NA) induced strong contraction. The thromboxane (TP) receptor antagonist ICI 192605 induced weak vasodilation, and this effect was significantly enhanced after precontraction of the vessels with uridine-5'-triphosphate (UTP). Incubation of BA segments with ICI 192605 attenuated the contractile effect of UTP. These data indicate that nNOS is not involved in resting cerebrovascular NO production and that basal TXA2 release induces a weak contractile tone in the rat BA. Activation of P2U receptors by UTP appears to stimulate TXA2 release in these vessels. PMID- 9736297 TI - Effect of L-arginine on adrenal and renal blood flows during hemorrhage in cats. AB - Our earlier studies have shown development of endothelial dysfunction in the feline renal artery during hemorrhagic hypotension. Because L-arginine (L-Arg), the precursor of nitric oxide (NO), reportedly improves endothelial function in several pathophysiological states including hypotension, we investigated its possible beneficial effect on the adrenal and renal circulations during hemorrhagic hypotension in anesthetized, ventilated cats. Hypotension (mean arterial pressure 50 mm Hg) significantly increased vascular resistance and decreased blood flow (radiolabeled microspheres) in both adrenal and renal cortices. L-Arg (30 mg/kg bolus, 10 mg/kg/min infusion, i.v.) had no significant hemodynamic effects in normotension but prevented the increase of the vascular resistance and improved blood flow in the adrenal cortex during hypotension. In the kidney, L-Arg also prevented hemorrhage-induced vasoconstriction, although its effect on blood flow did not reach significance. The NO synthase inhibitor N(G)-nitro-L-arginine (30 mg/kg bolus, 1 mg/kg/min infusion, i.v.) increased adrenal and renal vascular resistances to a similar extent as that observed during hypotension. It thus seems that an L-Arg-reversible dysfunction of the endothelial NO-synthesizing pathway contributes to hemorrhage-induced adrenal and renal vasoconstriction. PMID- 9736298 TI - Endothelin-1-induced contraction in cerebral vessels mediated by phospholipase C/protein kinase C cascade. AB - Endothelin (ET) vasoconstricts cerebral vessels potently, an effect mediated by ET(A) receptors on the smooth muscle, although the subsequent signaling cascade is unclear. We tested whether the action of ET-1 is mediated by the phospholipase C (PLC)/protein kinase C (PKC) cascade. Isometric force was measured in vitro in ring segments of rat basilar (BA) and middle cerebral (MCA) arteries and expressed as a percentage of the contraction to 124 mM K+. Concentration-effect curves for the constrictor effect of ET-1 (1 pM = 0.3 microM) in control segments or after 25 minutes preincubation with an inhibitor of PLC (neomycin 100 microM) or PKC (H7 10 microM) were constructed under resting tone. In untreated BA, 100 nM ET-1 induced a contraction of 119 +/- 5.3% that fell significantly to 97 +/- 2.8% and 98 +/- 6.7% after neomycin or H7 pretreatment, respectively. In MCA, 100 nM ET-1 induced a contraction of 105 +/- 3.2% that fell significantly to 93 +/- 6.3% and 64 +/- 8.1% after neomycin or H7, respectively. There was no significant shift of the ET-1 EC50 after PKC inhibition in either vessel or PLC inhibition in BA. In summary, the amplitude of ET-1-induced contraction in cerebral vessels is reduced significantly, whereas the sensitivity to the agonist is unchanged, after blocking PLC with neomycin or PKC with H7. This indicates noncompetitive inhibition. ET-1-induced contraction in cerebral vessels thus depends on activation of the PLC/PKC cascade. PMID- 9736299 TI - Dilator effect of bradykinin and acetylcholine in cerebral vessels after brain lesion. AB - Vasodilation elicited by bradykinin (BK) or acetylcholine (Ach) (10 nM-10 microM) in isolated rat cerebral arteries was studied under control conditions, after sham treatment, and after cold lesion (placing a cooled metal probe on the exposed dura) of the cortex. After 24 or 48 hours, isometric force was measured in ring segments of basilar (BA) and middle cerebral arteries (MCA). Concentration-effect curves were constructed after precontraction with 100 microM uridine triphosphate (MCA) or 1 microM serotonin (BA). In MCA and BA, BK elicited similar relative relaxations with maxima of 40.9 +/- 1.5% and 40.7 +/- 3.1%, respectively, at 1 microM. Ach-induced relaxation in BA was much stronger with 82.0 +/- 5.8% at 1 microM. MCA did not relax consistently to Ach. Relaxation to BK in MCA segments was not different between sham-treated and untreated animals. After cold lesion, the dilation to BK (1 microM) was significantly reduced at 24 hours from 0.7 +/- 0.06 to 0.4 +/- 0.06 mN. At 48 hours, this decrease was partly reversed (to 0.5 +/- 0.07 mN). In BA, there was no difference in Ach-induced relaxation between cold-lesioned or sham-treated animals. In summary, the nitric oxide (NO)-mediated response to BK in MCA is attenuated 24 hours after cold lesion. This damage to the BK/NO system is partly reversed 48 hours after the lesion. PMID- 9736300 TI - Control of rat glomerular microcirculation by juxtaglomerular adenosine A1 receptors. AB - The role of adenosine A1 receptors in the glomerular microcirculation and tubuloglomerular feedback (TGF) was studied in anesthetized Sprague-Dawley rats. TGF activity was assessed as the reduction in proximal tubular stop-flow pressure (SFP) on establishing orthograde perfusion of the loop of Henle with artificial tubular fluid at 40 nl/min. Administration of a selective A1 receptor antagonist, KW-3902 (0.5 microg/kg/min i.v.), increased fractional excretion of Na (FE(Na)) 4.3-fold without changing blood pressure, glomerular filtration rate, renal plasma flow, or filtration fraction. SFP in the absence of distal flow (SFP0) increased, and TGF-mediated SFP reduction was suppressed dose dependently [by 23 +/- 2% from an SFP0 of 34 +/- 1 mm Hg, by 15 +/- 4% from 36 +/- 2 mm Hg, and by 2 +/- 1% from 39 +/- 1 mm Hg during vehicle, low- and high-dose infusions (0.5 and 5.0 microg/kg/min), respectively]. Intratubular or peritubular capillary administration of 10(-4) M KW-3902 completely suppressed TGF without affecting SFP0. TGF suppression and elevation of SFP0 during systemic A1 blockade indicated vasodilation, both in the afferent arteriole and more proximal preglomerular vessels. Inhibition of tubular Na reabsorption combined with TGF suppression allowed the marked natriuresis. TGF suppression through systemic, luminal, and peritubular application of the drug suggest that juxtaglomerular apparatus A1 receptors are important in the control of glomerular microcirculation. PMID- 9736301 TI - The K(ATP) channel opener nicorandil: effect on renal hemodynamics in spontaneously hypertensive and Wistar Kyoto rats. AB - Adenosine triphosphate-sensitive K channels (K(ATP)) may subserve vasodilation in the renal microvasculature. Using micropuncture techniques, we examined whether the renal vasomotor action of K(ATP) differs in hypertensive and normotensive animals. Nicorandil (NC), a K(ATP) opener, was given i.v. (1 mg/hr/kg) to spontaneously hypertensive rats (SHR) or Wistar Kyoto rats (WKY). Mean arterial blood pressure decreased in both groups. Renal blood flow was almost unchanged in SHR but increased significantly in WKY. This effect was completely abolished by pretreatment with glibenclamide (GC; 3 mg/kg i.v.). To investigate the effect of NC on the regulatory mechanism of renal microcirculation, we measured proximal tubular stop-flow pressure (SFP) and assessed tubuloglomerular feedback (TGF) by monitoring SFP during loop perfusion with artificial tubular fluid. NC significantly increased SFP in WKY, an effect abolished by pretreatment with GC. In SHR, however, treatment with NC elicited no significant change in SFP. TGF response was not affected by NC treatment in either group. The data suggest that K(ATP) may modulate preglomerular vascular resistance, especially in the larger vascular segments not subject to TGF regulation, and may be attenuated in SHR. This might, at least in part, be attributable to the increased vascular resistance in SHR. PMID- 9736302 TI - Role of eicosanoids in renal angiotensin II vasoconstriction during nitric oxide blockade. AB - Nitric oxide (NO) buffers the effect of vasoconstrictors currently active in the renovascular system. Enhancement of the angiotensin II (Ang II)-induced vasoconstriction during NO blockade comprises both AT2-sensitive potentiation, decreasing the half maximal vasoconstriction (EC50) value to the subnanomolar concentration range, and augmentation, increasing the maximal effect (Emax) value in the isolated perfused rat kidney. In this study, we examine whether constrictory prostanoids are involved in Ang II subtype receptor (AT2)-sensitive potentiation of the Ang II effect during NO blockade. Thus, Ang II-induced vasoconstriction (0.1 or 10 nM Ang II) was measured in six series of constant flow perfused isolated rat kidneys in the presence of indomethacin under control conditions, during NO inhibition, and during combined inhibition of NO and all arachidonic pathways by eicosatetraynoic acid (ETYA), an analog of arachidonic acid. The vasoconstriction elicited by 10 nM Ang II, which is the maximal response, increased about threefold during NO inhibition compared with control. This augmentation was not affected by ETYA. In contrast, the vasoconstriction elicited by 0.1 nM Ang II increased about 20-fold during NO inhibition, reflecting mainly potentiation of the Ang II effect. This increase was abrogated by ETYA. We conclude that vasoconstrictor eicosanoids, which are suppressed by endogenous NO, mediate AT2-sensitive potentiation of the Ang II-induced vasoconstriction in the rat kidney. PMID- 9736303 TI - Influence of ketanserin on experimental loss of renal blood flow autoregulation. AB - Serotonin is important for effective renal blood flow (RBF) autoregulation in the normal rat and at two or seven days of reperfusion following renal ischemia. It has been suggested that after these reperfusion periods and during renal perfusion pressure (RPP) lowering, the vasodilatory autoregulation mechanism is not damaged but overwhelmed by increased 5-HT2-mediated vasoconstriction, resulting in complete loss of autoregulation. This study analyzes the influence of the 5-HT2-antagonist ketanserin on RBF autoregulation after two hours or one day of renal reperfusion following ischemia and in a model of cyclosporine (20 mg/kg.day for 10 days)-induced nephrotoxicity. Autoregulation was lost both after brief reperfusion periods and after cyclosporine. Similar to the two or seven days of reperfusion experiments, ketanserin in the cyclosporine model led to reappearance of autoregulation down to RPP 95 mm Hg. Despite an increased response to intrarenal serotonin after two hours of reperfusion, autoregulation was not restored by ketanserin. At one day of reperfusion and with ketanserin, autoregulation was present down to 105 mm Hg. Thus, during the early reflow period, other factors (of decreasing importance) most likely add to autoregulation loss. PMID- 9736304 TI - Is cerebral control of plasma [Na] a major determinant for systemic sodium balance? AB - To evaluate the role of volume expansion for prandial/postprandial natriuresis, we first determined spontaneous daily NaCl, H2O, and diet turnover and Evans blue and inulin spaces in male Wistar rats on various high-salt diets. Second, we measured the time course of Na and water clearance in chloralose/ketamine anesthetized rats over 270 minutes after a single intragastric Na load (0, 290.4, or 581 micromol/100 g body weight). Finally, similar measurements were made during and after a local [NaCl] increase in the left carotid artery supplying the brain for 60 minutes. Daily NaCl, H2O, and diet intake per rat was 2 to 74 mmol, 13 to 223 ml, and 1.5 to 33 g, respectively. Only inulin space and plasma [Na] correlated with daily Na uptake (X; regressions Y = 0.02X + 15.13, N = 99, r2 = 0.0716, P = 0.02; and Y = 141.7 + 0.1005X, N = 179, r2 = 0.104, P < 0.0001, respectively). Under chloralose/ketamine anesthesia, 86% to 102% of the total (i.v. plus i.g.) Na load and some 50% of the unilaterally administered intracarotid Na were excreted. Chloralose/ketamine anesthesia is thus suitable for studies on Na balance mechanisms. Plasma [Na] is under cerebral control. Because of its immediate onset, this mechanism might be the principal determinant of prandial and postprandial natriuresis and hence for the systemic Na balance. PMID- 9736305 TI - Sodium balance and blood pressure response to salt ingestion in uninephrectomized rats. AB - The possible role of extracellular volume (ECV) expansion in prandial/postprandial natriuresis was evaluated in control, sham-operated (SO), and uninephrectomized (UNX) male Wistar rats fed a 0.64 (normal salt, NS) or 8 (high salt, HS) g% NaCl diet for seven days after UNX. We thus determined daily NaCl, diet, and water intake and Evans blue and inulin spaces on day 7. Finally, we determined Na and water clearance after a single i.g. Na load (581 micromol/100 g body weight) under chloralose/ketamine anesthesia in UNX and control HS rats. NaCl, diet, and water intakes were comparable beyond day 5. Plasma volume and ECV were similar in all groups. With NS diet, glomerular filtration rate (GFR) in UNX was compensated but lower than that of SO rats (0.55 vs. 0.74 ml/min per 100 g body weight). Blood pressure (BP) was 111 mm Hg in SO controls and 112 mm Hg in the UNX group. After oral Na loading, BP rose in both groups and remained higher in UNX (134 vs. 126 mm Hg at 15 minutes, 130 vs. 118 mm Hg at 225 minutes). Cumulative Na and water excretions were similar (513 and 610 micromol/100 g body weight, 1.97 and 2.35 ml/100 g body weight in SO and UNX, respectively). Chronically salt-loaded UNX rats seem to maintain dietary Na balance by mechanism(s) other than volume expansion. PMID- 9736306 TI - Lipid peroxidation in the reperfusion injury of the liver. PMID- 9736307 TI - 4-Hydroxynonenal and cell signalling. PMID- 9736308 TI - Dynamics of vitamin E action against LDL oxidation. AB - Vitamin E acts as an important antioxidant against oxidative modification of low density lipoprotein (LDL) which is accepted as an initial event in the pathogenesis of atherosclerosis. In spite of the numerous studies and reports, the action and role of vitamin E have not been fully elucidated yet. In this brief overview, the dynamics of action of vitamin E as an antioxidant have been discussed and it is emphasized that the total antioxidant potency is determined by the relative importance of many competing reactions which is determined by the reactivities and concentrations of substrates, radicals and antioxidant and by physical factors of the environment. PMID- 9736309 TI - Lipid oxidation products and vascular function. AB - The endothelium and blood platelets are intimately involved in both the maintenance of vascular tone and in haemostasis. They are also exposed to high concentrations of lipoproteins, either in the plasma or in the sub-endothelial region of the artery wall, and the biological activity of these cells has been shown to be modulated. Oxidative modification of these lipoproteins results in further variations in the properties of these particles in relation to the activities of the endothelium and platelets. These effects and how the work of Hermann Esterbauer on the details of lipoprotein oxidation permitted rapid progress in understanding these phenomena are discussed in this review. PMID- 9736310 TI - Implications of lag time concept in the oxidation of LDL. AB - Oxidation of low density lipoprotein (LDL) has been implicated in the pathogenesis of atherosclerosis. The most common technique for measuring the oxidation of lipoproteins is the continuous measurement of the formation of conjugated diene at OD 234 nm. The concept of "lag time", derived from such measurements, has been used to test the efficacy of various antioxidants for their ability to inhibit the oxidation of LDL. This review will elaborate on some of the factors that might affect the lag time. PMID- 9736311 TI - Nitric oxide and low-density lipoprotein oxidation. AB - Nitric oxide can have both pro-oxidant and antioxidant effects on low-density lipoprotein. Nitric oxide does not appear to react directly with components of LDL. However, in the presence of oxygen (through NO2 and N2O3 formation) or superoxide (through peroxynitrite formation) nitric oxide may cause oxidation of the lipid, protein and antioxidant components of LDL. Conversely, nitric oxide is a potent inhibitor of LDL oxidation when initiated by copper ions or by azo initiators. The possible implications of these observations to vascular pathology are discussed. PMID- 9736312 TI - The lag phase. AB - Peroxidation of lipids in membranes and lipoproteins proceeds through the classical free radical sequence encompassing initiation, propagation, and termination phases which are expressed by a lag phase in which little oxidation occurs, followed by a rapid increase in autocatalysis by chain-propagating intermediates and, finally, a decrease in the rate of oxidation. The lag phase is lengthened by preventive or chain-breaking antioxidants, which scavenge the initiation reaction or intercept the chain-carrying species. Hence, the lag phase in lipid peroxidation processes reflects the antioxidant status of membranes and lipoproteins and, as a corollary, their resistance to oxidation. A large number of lipid peroxidation studies with different membranes attest to the complex free radical network underlying this process. The type of initiator and the steady state level of oxygen are important factors that affect differently the rates of the individual steps of peroxidation. Equally complex are the factors that influence the lag phase preceding the oxidation of LDL. Lipid peroxyl radicals play a key role in the dynamics of lipid peroxidation: on the one hand, the lag phase is best defined for chain-breaking compounds able to reduce peroxyl radicals; on the other hand, the overall time course of lipid peroxidation is largely influenced by the rate constants for propagation reactions and termination involving peroxyl radical recombination. PMID- 9736313 TI - Cellular thiol production and oxidation of low-density lipoprotein. AB - Compelling evidence suggests that low-density lipoprotein (LDL) is oxidized by cells within the arterial intima and that, once oxidized, it is profoundly atherogenic. The precise mechanism(s) by which cells promote the oxidation of LDL in vivo are not known; in vitro, however, oxidation of LDL can be enhanced by a number of differing mechanisms, including reaction with free and protein-bound metal ions, thiols, reactive oxygen species, lipoxygenase, myeloperoxidase and peroxynitrite. This review is concerned with the mechanisms by which cells enhance the oxidation of LDL in the presence of transition metals; in particular, the regulation, pro- and anti-oxidant consequences, and mechanism of action of cellular thiol production are examined, and contrasted with thiol-independent oxidation of LDL in the presence of transition metals. PMID- 9736314 TI - Lipid peroxidation and biogenic aldehydes: from the identification of 4 hydroxynonenal to further achievements in biopathology. AB - The formation, reactivity and toxicity of aldehydes originating from lipid peroxidation of cellular membranes are reviewed. Very reactive aldehydes, namely 4-hydroxyalkenals, were first shown to be formed in autoxidizing chemical systems. It was subsequently shown that 4-hydroxyalkenals are formed in biological conditions, i.e. during lipid peroxidation of liver microsomes incubated in the NADPH-Fe systems. Our studies carried out in collaboration with Hermann Esterbauer which led to the identification of 4-hydroxynonenal (4-HNE) are reported. 4-HNE was the most cytotoxic aldehyde and was then assumed as a model molecule of oxidative stress. Many other aldehydes (alkanals, alk-2-enals and dicarbonyl compounds) were then identified in peroxidizing liver microsomes or hepatocytes. The in vivo formation of aldehydes in liver of animals intoxicated with agents that promote lipid peroxidation was shown in further studies. In a first study, evidence was forwarded for aldehydes (very likely alkenals) bound to liver microsomal proteins of CCl4 or BrCCl3-intoxicated rats. In a second study, 4-HNE and a number of other aldehydes (alkanals and alkenals) were identified in the free (non-protein bound) form in liver extracts from bromobenzene or allyl alcohol-poisoned mice. The detection of free 4-HNE in the liver of CCl(4) or BrCC1(3)-poisoned animals was obtained with the use of an electrochemical detector, which greatly increased the sensitivity of the HPLC method. Furthermore, membrane phospholipids bearing carbonyl groups were demonstrated in both in vitro (incubation of microsomes with NADPH-Fe) and in vivo (CC1(4) or BrCCl(3) intoxication) conditions. Finally, the results concerned with the histochemical detection of lipid peroxidation are reported. The methods used were based on the detection of lipid peroxidation-derived carbonyls. Very good results were obtained with the use of fluorescent reagents for carbonyls, in particular with 3-hydroxy-2-naphtoic acid hydrazide (NAH) and analysis with confocal scanning fluorescence microscopy with image video analysis. The significance of formation of toxic aldehydes in biological membranes is discussed. PMID- 9736315 TI - Determinants of intestinal detoxication of lipid hydroperoxides. AB - It has long been recognized that hydroperoxides are agents of cytotoxicity. However, in recent years, it is increasingly apparent that lipid hydroperoxide may play an important role in mediating cellular and molecular events in degenerative pathophysiological processes that lead to intestinal disorders, such as cancer. Yet, surprisingly, little is known of the intestinal disposition of peroxidized lipids and of the metabolic factors that determine mucosal peroxide elimination. The present paper summarizes the evidence for the pivotal role of reductant (GSH and NADPH) availability in intestinal peroxide detoxication. This information will provide important insights into the relationship between luminal lipid hydroperoxides and intestinal GSH redox homeostasis, and is pertinent to understanding how dietary oxidants like lipid peroxides, can impact intestinal integrity with implications for genesis of gut pathology. PMID- 9736316 TI - Glutathione S-transferases and prevention of cellular free radical damage. AB - This paper describes the intervention of glutathione-dependent enzymes, in particular the glutathione S-transferases (GSTs), in both the detoxication of electrophilic decomposition products resulting from the attack of oxygen radicals on lipids and DNA; and the prevention of oxygen toxicity generated by redox cycling catecholamine derivatives. The continuing growth of our knowledge of the glutathione S-transferase polygene family is described in terms of the increase in members of known gene families, the discovery of new ones and our increasing knowledge of their activities towards endogenous substrates. PMID- 9736317 TI - Measurement of lipid peroxidation. AB - Lipid peroxidation results in the formation of conjugated dienes, lipid hydroperoxides and degradation products such as alkanes, aldehydes and isoprostanes. The approach to the quantitative assessment of lipid peroxidation depends on whether the samples involve complex biological material obtained in vivo, or whether the samples involve relatively simple mixtures obtained in vitro. Samples obtained in vivo contain a large number of products which themselves may undergo metabolism. The measurement of conjugated diene formation is generally applied as a dynamic quantitation e.g. during the oxidation of LDL, and is not generally applied to samples obtained in vivo. Lipid hydroperoxides readily decompose, but can be measured directly and indirectly by a variety of techniques. The measurement of MDA by the TBAR assay is non-specific, and is generally poor when applied to biological samples. More recent assays based on the measurement of MDA or HNE-lysine adducts are likely to be more applicable to biological samples, since adducts of these reactive aldehydes are relatively stable. The discovery of the isoprostanes as lipid peroxidation products which can be measured by gas chromatography mass spectrometry or immunoassay has opened a new avenue by which to quantify lipid peroxidation in vivo, and will be discussed in detail. PMID- 9736318 TI - Prolactin, a pituitary hormone that modifies immune responses. Proceedings of the Mini-symposium on Prolactin and SLE, held at the 5th International Conference on Systemic Lupus Erythematosus, Cancun, Mexico. PMID- 9736319 TI - Anti-endothelial cell antibodies in the sera of hyperprolactinemic women. AB - Prolactin (PRL) is closely associated with autoimmune diseases in animal models and humans, and several disease-related autoantibodies were reported in increased titers in patients with hyperprolactinemia (HPRL). We studied the presence of anti-endothelial cell antibodies (AECA) and other autoantibodies in sera of female patients with HPRL. Sera from 25 HPRL patients and 10 healthy female controls were tested for AECA (against both macrovascular and microvascular endothelial cell antigens), anti-dsDNA, and anti-cardiolipin (anti-CL) using ELISA. Sera were considered positive for the autoantibody when the optical density (OD) value was more than 3 s.d. above the mean of the OD in normal controls. Sera from 13 patients were obtained repeatedly during dopaminergic anti PRL treatment, to relate PRL level or anti-PRL treatment with the autoantibody levels. Elevated micro and/or macrovascular AECA were observed in sera from 19/25 patients (76%). Elevated titers of anti-CL Abs, all beta2-GPI-dependent, and low levels of anti-dsDNA antibodies (Abs) were also observed in the HPRL patients. Inhibition studies showed that the affinity purified AECAs bound the endothelial cell (EC) antigens in a dose-dependent manner. Titers of AECA as well as anti-DNA and anti-CL autoantibodies did not correlate with PRL level nor with the use or duration of anti-PRL treatment. None of the HPRL patients presented clinical manifestations of autoimmune disease. We conclude that elevated levels of AECA as well as anti-DNA and anti-CL autoantibodies are frequent in hyperprolactinemia. Our results further support the association of PRL and autoimmunity, and may point to a relationship between AECA-associated diseases and HRPL. The presence of autoantibodies in patients with HPRL might portend an increased risk for future development of autoimmune disease. PMID- 9736320 TI - Prolactin levels in patients with systemic lupus erythematosus: a case controlled study. AB - Recent accumulated evidence suggests that prolactin (PRL) is an important immunomodulator and might have a role in the pathogenesis of systemic lupus erythematosus (SLE). Our aim was to assess the frequency of hyperprolactinemia in women with SLE and to evaluate its correlation with disease activity. PRL plasma levels were measured in 36 women with SLE and 20 age-matched healthy controls. We excluded patients with renal and/or hepatic failure, pregnant patients and patients taking drugs which could increase PRL levels. Disease activity was assessed using the SLE disease activity index (SLEDAI). Patients with a score > 10 were considered active. In patients and controls, PRL levels were determined by radioimmunoassay (RIA) during the first part of the menstrual cycle (between the 5th and 8th day) (normal value < 20 ng/ml). Ten of 36 (27.7%) SLE patients had high PRL levels (> 20 ng/ml). The mean PRL level was higher in SLE than in the control group (17.1+/-12.9 s.d. vs 9.9+/-3.5, P < 0.01). Patients with active disease had a trend to higher mean PRL levels than inactive patients although this difference was not statistically significant (21.1+/-4.8 vs 14.8+/-6.9, P = 0.09). No correlation was found between PRL levels and SLEDAI score. Furthermore, no significant correlation was found between PRL levels and any clinical or serological finding. PMID- 9736321 TI - Clinical significance of serum and urine prolactin levels in lupus glomerulonephritis. AB - Prolactin (PRL) has been involved in the pathogenesis of systemic lupus erythematosus (SLE) and hyperprolactinemia has been connected with systemic activity. However, the clinical significance of PRL has not been investigated in lupus glomerulonephritis (GN). METHODS: We studied SLE patients (ACR criteria) with biopsy-proven renal disease. Renal histology was classified according to World Health Organization (WHO) criteria. Renal function tests, albuminuria, complement levels (nephelometry), anti-DNA antibodies (C. luciliae) and serum and urine PRL concentrations (RIA) were determined at baseline and at 4-month intervals for one year. Renal activity was defined as mild, moderate or severe according to serum creatinine, creatinine clearance, albuminuria, red blood cells (RBC), and casts. RESULTS: There were 26 patients with mean age 28.5 y and mean disease duration 47.9 months. Twenty patients had diffuse proliferative glomerulonephritis (GN), four had focal GN and two had membranous GN with proliferative changes. Renal activity was mild in ten patients, moderate in ten and severe in six. Mean serum (24.7+/-5.3) and urine (0.90+/-0.36) PRL levels were higher in patients with severe renal activity (P < 0.05 compared with mild group). PRL levels decreased after treatment, but this trend was not uniform during the follow-up period. CONCLUSION: Hyperprolactinemia was prevalent in SLE patients and high levels of PRL in the serum and urine could be related to severe renal disease. PMID- 9736322 TI - Plasma homovanillic acid and prolactin in systemic lupus erythematosus. AB - Prolactin (PRL) is an important immunoregulator and might have a role in the pathogenesis of systemic lupus erythematosus (SLE). The regulation of pituitary prolactin secretion is complex and involves a negative feedback process in the hypothalamus, in which dopamine plays the principal role. However, the main source of serum prolactin in lupus patients is still not clearly established. Since homovanillic acid (HVA), the principal metabolite of dopamine (DA), is removed from the brain into the blood, it would indirectly reflect DA metabolism. It is assumed that the turnover of a neurotransmitter can be determined through an analysis of its metabolites. The objective of this study was to analyse plasma samples from SLE patients to see if there were any alterations in neurally functioning DA through its principal metabolite, HVA. We also measured the levels of PRL and compared HVA and PRL with the clinical activity of the disease. Twenty four SLE patients and fifteen healthy controls were studied. The investigation was done over a period of 3 months. The results of this study show significantly low levels of HVA in lupus patients compared to controls (P < 0.0001). This corresponds to a decrease in dopamine turnover. Hyperprolactinemia was observed in nine patients, and the average level of prolactin in lupus patients was higher than in healthy controls (P < 0.001). For the duration of the study, a significant percentage of variation was observed in the levels of HVA in the clinically active patients (P < 0.05) compared to inactive patients. When PRL was compared in these groups, throughout the study, no significant percentage of variation was observed. The relationship between HVA and PRL in healthy controls was r = 0.47, P = 0.08, and in patients was r = 0.04, P = 0.84. It is suggested that there is a probable association between plasma levels of HVA and PRL in the healthy controls and not in the SLE patients. PMID- 9736323 TI - Anti-prolactin autoantibodies in systemic lupus erythematosus patients with associated hyperprolactinemia. AB - Hyperprolactinemia has been found in a subset of systemic lupus erythematosus (SLE) patients. In order to explore whether antibodies to prolactin (PRL) play a role in SLE patients with associated hyperprolactinemia, we performed a cross sectional study in which 259 consecutive SLE patients were tested for hyperprolactinemia and anti-prolactin autoantibodies. Forty-one (15.8%) had prolactin levels above the norm. The frequency of anti-PRL autoantibodies in hyperprolactinemia was 2/14 (14.3%). In the SLE patients with 'idiopathic hyperprolactinemia', 11/27 (40.7 %) had anti-PRL antibodies. The levels of serum PRL were significantly higher in patients with idiopathic hyperprolactinemia and anti-PRL autoantibody compared to the patients with idiopathic hyperprolactinemia who were anti-PRL autoantibody-negative. Patients with idiopathic hyperprolactinemia and anti-PRL autoantibody had relatively low SLE disease activity index (SLEDAI) scores and significantly different laboratory parameters compared to those with idiopathic hyperprolactinemia and no anti-PRL antibody. There was a significant correlation between titers of the anti-PRL autoantibody and serum PRL levels (rs = 0.98, P = 0.0001). These data suggest that anti-PRL antibodies could be the cause of hyperprolactinemia in a subset of SLE patients, especially those with particularly high serum prolactin levels with a diagnosis of 'idiopathic hyperprolactinemia'. The patients with anti-PRL antibody had fewer clinical manifestations and less serological activity, indicating that biological activity of PRL was attenuated by the autoantibody. PMID- 9736324 TI - Hypothalamic-pituitary-adrenal axis function and prolactin secretion in systemic lupus erythematosus. AB - The objective was to study the response of cortisol and of prolactin (PRL) to specific stimuli in systemic lupus erythematosus (SLE). We measured the response of cortisol to insulin-induced hypoglycemia and of PRL to thyrotropin releasing hormone (TRH) in seven patients with active untreated SLE and in ten paired control subjects. All were women with regular menstrual cycles. With the exception of two patients, they had never received corticosteroids before the study. The basal serum levels of cortisol (12.5+/-2.4 microg/dl) and PRL (10.7+/ 1.0 ng/ml) in the SLE group were not significantly different from those of the control group (12.3+/-1.1 microg/dl and 13.7+/-2.4 ng/ml, respectively). The cortisol response after hypoglycemia was significantly lower in SLE patients compared to the control group at 45 min (P=0.01), at 60 min (P = 0.009), and at 90 min (P = 0.001). The integrated cortisol response to hypoglycemia expressed as area under the response curve (AUC) did not differ significantly in either group (1447+/-187 vs 1828+/-84, P = 0.06). Although the peak of PRL after TRH did not differ significantly in both groups (68.0+/-7.4 ng/ml in SLE vs 66.3+/-77 ng/ml in controls) and the AUC of PRL response after TRH was comparable in both groups (4672+/-537 vs 4128+/-541, P = 0.32), the interval-specific 'delta' response was significantly higher in SLE than the control group at 0-60 min (P=0.02) and 0-90 min (P = 0.01) after TRH injection. These findings suggest that active SLE is associated with some degree of dysfunction of the hypothalamic-pituitary glucocorticoid axis and PRL secretion. PMID- 9736325 TI - The hypothalamic-pituitary response in SLE. Regulation of prolactin, growth hormone and cortisol release. AB - It has been suggested that neuroendocrine regulation plays an important role in the pathogenesis and activation of autoimmune diseases. The aim of this investigation was to clarify the hypothalamic-pituitary response to a well defined stimulus under standardised conditions in patients with SLE. Plasma concentrations of prolactin (PRL), growth hormone (GH) and cortisol were determined in venous blood drawn through an indwelling cannula during insulin induced hypoglycaemia (0.1 U/kg b.w., i.v.) in ten patients and in 12 age-, gender- and weight-matched healthy subjects. Basal PRL concentrations were higher in patients vs healthy controls (12 vs 6 ng/ml, P < 0.01), though still within the physiological range. Insulin-induced plasma PRL and GH were significantly increased both in patients and healthy subjects; however, the increments or areas under the curves were not different in the two groups. Plasma cortisol response showed moderate attenuation in patients. Sensitivity of pituitary lactotrothrops to thyrotropin-releasing hormone (TRH) administration (200 microg, i.v.) was the same in patients and control subjects. In SLE patients with low activity of the disease the sensitivity of pituitary PRL release to TRH administration remained unchanged. The hypothalamic response to stress stimulus (hypoglycaemia) was comparable in patients and healthy subjects. PMID- 9736326 TI - Bromocriptine in systemic lupus erythematosus: a double-blind, randomized, placebo-controlled study. AB - The objective of this study was to investigate the efficacy and safety of bromocriptine (BRC) as an adjunct to conventional treatment in systemic lupus erythematosus (SLE). A prospective, double-blind, randomized, placebo-controlled study compared BRC at a fixed daily dosage of 2.5 mg with placebo. Patients were followed for 2-17 months (mean 12.5 months). Disease activity was assessed using the SLE Disease Activity Index (SLEDAI), numbers of flares were recorded, and serum prolactin (PRL) levels were obtained at intervals during the study. Patients were allowed to take prednisone and immunosuppressive drugs. Sixty-six patients with SLE entered the study. Thirty-six were treated with BRC, and 30 controls received placebo. Sixteen patients were removed from the study during the treatment period: five in each group left the study because of adverse effects, five became pregnant, and one patient who took placebo died with central nervous system lupus. Four patients in the BRC treatment group and three patients in the placebo group moved away or stopped coming for study visits for unknown reasons, and were lost to follow-up during the course. At entry, serum PRL was (mean+/-s.d.) 24.8 ng/ml+/-18.4 in the BRC treatment group. This value fell to 5.8+/-9.0 after 12 months of treatment. Corresponding PRL values in controls were 23.7+/-22.1 pretreatment and 20.3+/-14 after 12 months. PRL levels in BRC-treated subjects were significantly lower than levels in control subjects after 3, 6, 9, and 12 months of treatment. The SLEDAI score on the fifth protocol visit was decreased significantly in the BRC group vs controls: 0.9+/-1.4 vs 2.6+/-4.5 (P < 0.05). Although the absolute number of flares in each group was similar, the mean number of flares/patient/month was decreased significantly in the BRC group compared to the control group (0.08+/-0.1 vs 0.18+/-0.2, P = 0.03). Long term treatment with a low dose of BRC appears to be a safe and effective means of decreasing SLE flares in SLE patients. PMID- 9736327 TI - Differential effects of estrogen and prolactin on autoimmune disease in the NZB/NZW F1 mouse model of systemic lupus erythematosus. AB - Estrogen and prolactin have been shown to modulate autoimmunity in the NZB/NZW F1 (B/W) mouse model of systemic lupus erythematosus (SLE). However, estrogen stimulates prolactin secretion. The goal of this study was to examine differential effects of estrogen and prolactin in the female B/W mouse model of SLE. B/W females were manipulated to create combinations of low and high concentrations of serum estrogen and prolactin. Hyperprolactinemic mice with either low or high serum estrogen levels had accelerated development of albuminuria at 24 and 32 weeks of age compared to normal and hypoprolatinemic mice. High estrogen/high prolactin mice also had a higher percentage of anti-DNA antibodies compared to mice in the low estrogen/low prolactin and the high estrogen/low prolactin groups. IgG levels were not significantly different between groups. Mean survival was shortest in the high estrogen/high prolactin group (34+/-1.0 weeks) and longest in the high estrogen/low prolactin group (42+/ 1.2 weeks; P < 0.05). High levels of serum estrogen were associated with depressed in vitro lymphoproliferation and IL-2 production. This study suggests that high prolactin levels in either high or low serum estrogen states are associated with accelerated autoimmunity in the B/W mouse. This study further demonstrates that high estrogen levels do not accelerate murine SLE when the prolactin-stimulating property of estrogen is suppressed by bromocriptine. Further investigation of hormonal interactions in autoimmunity will provide a better understanding of hormonal immunoregulation and, perhaps, lead to improved clinical application of hormonal immunomodulation. PMID- 9736328 TI - On the prevalence of homosexuality and bisexuality, in a random community survey of 750 men aged 18 to 27. AB - A stratified random sample of 750 males aged 18 to 27 in Calgary, Canada included questions on sexual activity and orientation. A computerized response format (established as a good method for eliciting sensitive personal data) ensured anonymity. Three measures of homosexuality were employed: (1) voluntary, same gender sexual contact from age 12 to 27: 14.0%; (2) overlapping homosexual (5.9%) and/or bisexual (6.1%) self-identification: 11.1%; and (3) exclusive (4.3%) and non-exclusive (4.9%) same-gender sexual relationships in past 6 months: 9.2%. On the basis of one or more of the three often overlapping measures, 15.3% of males reported being homosexual to some degree. CES-D depression scores did not differ significantly for sexually active homosexual (mean 14.6), bisexual (mean 15.7), and heterosexual (mean 13.7) males. The elevated depression scores for celibate homosexual (mean 27.1) and heterosexual (mean 23.6) males permit various interpretations, but are not supportive of beliefs and related institutional policies recommending or requiring that young adult homosexual males be celibate. PMID- 9736329 TI - S/M (sadomasochistic) interactions in semi-public settings. AB - An organized semi-public event for the exhibition of S/M (sadomasochistic) behavior is known as a "party" by the participants. Using a retrospective analysis of the author's experiences over the last 25 years, a description of these parties is presented. The present paper explores the structure, function, and purpose of these parties. The S/M behavior, sexual interactions, rules of etiquette, and structure of the party are summarized. The lack of genitally focused orgasm-seeking behavior at the party is discussed at length. PMID- 9736330 TI - Accepting the gay person: rental accommodation in the community. AB - Effects of the "homosexual" label in obtaining community accommodation were examined, in a sample of 180 individuals advertising rooms or flats for rent in two Canadian cities, Windsor and London, Ontario, and in Detroit, Michigan. Telephone calls, for half the sample, made simple enquiries as to availability; for the other half, similar enquiries were made by an individual who was ostensibly homosexual. In the latter condition, rooms were significantly more likely to be described as unavailable. Comparisons are made to similar, previous research, and to current perspectives about community reactions to stigmatizing conditions. PMID- 9736331 TI - This powerful opening of the heart: how ritual affirms lesbian identity. AB - This study was undertaken to examine the ways in which ritual helps affirm lesbian identity. The study explored two types of rituals commonly used among lesbians: commitment ceremonies and baby naming or child dedication ceremonies. Six in-depth interviews were conducted with lesbians who had participated in rituals which they perceived as affirming to their identities. The major findings showed that the respondents were in significant agreement that their rituals helped solidify lesbian identity through the confrontation of internalized and external homophobia. Subjects reported that this process subsequently fortified them in their daily battles against oppression. Of further significance, the respondents all specified that in several ways, their ritual helped provide balance and establish equilibrium between the many disparate experiences that result from living in a homophobic society. Implications will specify ways that therapists can use ritual as a tool in helping lesbian clients solidify and affirm their identities. PMID- 9736332 TI - An experiential attitude change: social work students and homosexuality. AB - This paper describes and analyzes an attitude change among third year undergraduate social work students in a major university in Israel. The subject of attitudes towards homosexuality, while receiving extensive theoretical attention, lacks documentation of actual change in attitudes. This paper is an attempt to fill the gap by using a quasi-experimental non-equivalent control group design evaluation of the impact of an elective course on homosexuality on students' attitudes. The experimental group was comprised of 31 students who enrolled in an elective course, entitled "Individual, familial, and social aspects of homosexuality." The course combined theoretical as well as experiential frameworks. The control group was comprised of 56 third-year social work students who did not enroll in the course. Both quantitative and qualitative methodologies were employed in comparing the two groups prior to and following the intervention. Specifically, a 25-item questionnaire was used to measure homophobia attitudes, and an open-ended question was used to qualitatively explore the students' associations with the term "homosexuality." Findings are discussed within the frameworks of the current literature regarding attitudes towards homosexuality, attitude change, social work education, and the unique situation in Israel. PMID- 9736333 TI - Helping families with homosexual children: a model for counseling. AB - Adolescence is a stressful time for many children and identification as homosexual adds to the frustration normally experienced during this period of development. The processes of coming out and disclosing this homosexual identity, while often difficult, are necessary if the adolescent is to develop a stable sexual identity. In families where disclosure has occurred, feelings of regret, confusion, and denial are common; in order to deal with these feelings, many families seek professional help. It is important the therapist working with these family systems has an understanding of homosexuality, positive attitudes towards homosexuality, and an appropriate counseling model to use with these family systems. A counseling model based on problem-solving communication-training (Robin & Foster, 1984, 1989), meant as a framework upon which professionals can build, is presented; professional attitudes, assessment processes, and goal setting are also discussed. PMID- 9736334 TI - "Squalid with joy": scobie, sex, and race in Lawrence Durrell's Alexandria quartet. AB - Edward Said and others have identified the Orient as a space of sexual opportunity for western travelers and servants of empire, while more recently Joseph Boone and Rudi Bleys hav focused on, respectively, the "homerotics of orientalism" and the "motives and conditions of homosexual exile." In terms of both sexual and imperial politics, Lawrence Durrell's Scobie, "peddyrast" and transvestite, presents a challenge. I have chosen to rehabilitate rather than deconstruct this determinedly transgressive figure, perhaps the most oddly privileged of all the characters in the Alexandria Quartet. Though Scobie may exhibit some orientalizing and colonial habits of mind, his more positive gestures, such as his relationship with Abdul, are unique in this sequence of novels. Within the limits of his time and generation, and within the confines of an imperial culture, he comes closest to reconciling East and West as he destabilizes gender roles and challenges racial division. PMID- 9736335 TI - Inhibition of caspase activity prevents anti-IgM induced apoptosis but not ceramide generation in WEHI 231 B cells. AB - In apoptosis induced by Reaper in Drosophila, as well as in a number of other systems, it has been suggested that the increased synthesis of ceramide might be a consequence of the activation of the caspase/ICE (Interleukin-1beta converting enzyme) protease pathway involved in cell death, implying that ceramide generation might often be the result rather than the cause of apoptosis. WEHI 231 B cells have previously been shown to undergo apoptosis following exposure to exogenous ceramide and to produce increased amounts of ceramide in response to anti-IgM crosslinking. We show here that in WEHI 231 cells a peptide inhibitor of caspase activity blocks cell death in response to both anti-IgM and exogenous ceramide. However, the induction of ceramide synthesis by WEHI 231 cells in response to anti-IgM crosslinking is not blocked by this peptide. These results indicate that antigen receptor induced ceramide generation in WEHI 231 cells does not require caspase activation, and support the view that ceramide generation in immature B cells may be the cause rather than the consequence of activation of the caspase dependent death pathway. PMID- 9736336 TI - The association between CD20 and Src-family Tyrosine kinases requires an additional factor. AB - CD20 is a B cell surface protein which can initiate intracellular signals involving tyrosine kinase activation, and modify B cell growth and differentiation. CD20 is tightly associated with the Src-family kinases Lyn, Fyn and Lck; however, the mechanism of interaction remains to be determined. Association between CD20 and Src-family kinases has been detected in peripheral blood B cells and in 5 out of 8 unrelated B cell lines. The lack of CD20 associated kinase activity in some cell lines offered an opportunity to investigate the mechanism of CD20 associations. All 8 B cell lines were found to express Lyn, and, with one exception, all cell lines also expressed Fyn. Lck, however, was not detected in any of the cell lines in which CD20 failed to coprecipitate kinase activity. To test the possibility that Lck was required for assembly of the CD20 complex, Lck was transfected into one of the 3 CD20/kinase association-deficient lines, namely T51. CD20 did not coprecipitate kinase activity from the transfected T51 cells, despite their expression of high levels of exogenous Lck, as well as endogenous Lyn and Fyn. CD20 cDNA from T51 was sequenced and found to be normal. These data establish that association between CD20 and Src-family kinases requires an additional factor. PMID- 9736337 TI - Identification of conserved amino acids in murine B7-1IgV domain critical for CTLA4/CD28:B7 interaction by site-directed mutagenesis: a novel structural model of the binding site. AB - The B7: CD28/CTLA4 interaction plays a major role in T cell responses. Immune intervention targeted at this interaction has demonstrated a vast potential in enhancing tumor immunity and blocking autoimmunity and transplant rejection. However, the structural basis for this interaction is unclear. While we and others have performed site-directed mutagenesis to define amino acids involved in binding CD28 and CTLA4, these residues are localized in different regions, and it is unlikely for all of them to be directly involved. In addition, the effect of the mutations on the overall conformation of B7 has not been systematically evaluated. In this study, we have carried out site-directed mutagenesis to define the amino acids within B7-1 IgV-like domain which participate B7:CD28/CTLA4 interaction. Four anti-B7-1 mAbs that recognize three independent antigenic epitopes on B7-1 were used to monitor the effect of mutations on the overall conformation of B7-1. Of the five mutations in the IgV domain that we have produced, D113 > A appears to interfere with cell surface expression and/or overall conformation of B7-1. while four others do not significantly affect the overall conformation and cell surface expression of B7-1. Among them, G115 > A and Y91 > A eliminated B7-1 binding to both CD28Ig and CTLA4Ig; our previously reported mutants L109 > A and W88 > A selectively affect the B7-1 binding to either CD28Ig or CTLA4Ig. Structural modeling of B7-1 based on the structure of immunoglobulin revealed that these four and other previously identified critical amino acids in both IgV- and IgC-like domains can form a localized structure. PMID- 9736338 TI - V(H) replacement is unlikely to contribute significantly to receptor editing due to an ineffectual embedded recombination signal sequence. AB - Receptor editing is a process consisting of replacement of pre-existing H or L chain rearrangements by secondary rearrangements. This process could serve to remove autoreactive specificities, or to rescue loci with non-functional rearrangements. At the H chain locus, functional replacement of a V(H)DJ(H) rearrangement by an upstream V(H) requires the presence of an embedded RSS located in reverse orientation near the 3' end of the V(H) segment. Although most V(H) genes contain a fairly consensus embedded heptamer, the nonamer sequence bears little resemblance to the consensus RSS nonamer. Therefore, the physiologic rate of H chain editing by V(H) replacement is yet unknown. In this study, we used both conventional and sensitive competition recombination substrate assays to determine the recombination frequency of the V(H)1X embedded RSS relative to consensus and non-consensus RSS's. Results show no detectable recombination of the 81X embedded RSS in a recombination substrate, and the competition substrate allows us to estimate that the 81X embedded RSS recombines at least 1300 fold less often than a consensus RSS. This suggests that V(H) gene replacement is not responsible for the decrease in representation of the 81X gene during differentiation. Furthermore, since the sequence of the embedded RSS is very similar for many V(H) genes, our results suggest that receptor editing of the H chain will be an infrequent event, leaving L chain editing as the main mode of avoiding autoreactive specificities in vivo. PMID- 9736339 TI - Insertions and deletions in hypervariable loops of antibody heavy chains contribute to molecular diversity. AB - Antibody diversity, a molecular feature which allows these proteins to specifically interact with a diverse set of targets, is created at the genetic level by a variety of means. These include germline gene segment recombination, junctional diversity and single basepair (bp) substitution. We here demonstrate that a human high affinity antibody specific for an exogenous protein antigen carry three amino acid residues immediately adjacent to the first hypervariable loop of the heavy chain. These additional residues are shown not to be encoded by the germline repertoire. We also describe the characteristics of insertions and deletions, not found in any known germline sequence, within the first and second hypervariable loops of other previously described antibody-encoding genes. These findings demonstrate that insertions or deletions of entire codons provide yet another approach by which the human antibody repertoire is diversified in vivo. Since these major genetic modifications occur within or immediately adjacent to loops contributing to the antigen-binding site, they are likely to affect the binding properties of the mutated antibodies. PMID- 9736340 TI - Interferon-gamma-induced factor binding to the interleukin-4-responsive element of CD23b promoter in human tonsillar mononuclear cells: role in transient up regulation of the interleukin-4-induced CD23b mRNA. AB - Stimulation of human tonsillar mononuclear cells with interleukin-4 (IL-4) and interferon-gamma (IFN-gamma) rapidly induced the activation of distinct nuclear factors with different mobilities, both of which bind the IL-4 response element (IL-4RE) of CD23b promoter as examined by electrophoretic mobility shift assays (EMSA). Co-treatment of IL-4 and IFN-gamma induced, in addition to the two distinct complexes, a new complex with an intermediate mobility. The IL-4-induced complex reacted with anti-STAT (signal transducers and activators of transcription) 6, resulting in a supershift whereas the formation of the IFN gamma-induced complex was inhibited by anti-STAT 1. The intermediate complex appeared to react with both anti-STAT 6 and anti-STAT 1. Although IFN-gamma alone did not induce CD23 mRNA transcription, Northern blot analysis revealed a transient up-regulation of the IL-4-induced CD23 mRNA by IFN-gamma within 2 h of IFN-gamma treatment in these tonsillar cells. The results suggest that the IL-4RE of the IL-4-inducible gene can accommodate both IL-4- and IFN-gamma-activated factors, such as STAT 6 and STAT 1, either in homodimeric or heterodimeric forms and the binding of these different proteins to the respective promoter may play a potential regulatory role in the IL-4-inducible gene expression. PMID- 9736341 TI - Stimulation of the high-affinity IgE receptor results in the tyrosine phosphorylation of a 60 kD protein which is associated with the protein-tyrosine kinase, Csk. AB - The protein tyrosine kinase Csk downregulates the activity of the Src family of kinases and has a negative effect on signal transduction through several Src kinase-associated receptors. Because the Src-family kinase Lyn plays a pivotal role in FcepsilonRI-mediated cellular activation, we examined whether Csk is involved in FcepsilonRI signaling events. Using anti-Csk antibodies and recombinant fusion proteins we detected a single tyrosine-phosphorylated protein of 60 kD (herein referred to as 'p60') that associates with the SH2 domain of Csk after stimulation of the FcepsilonRI. p60 phosphorylation reached a maximum within one minute and remained constant while the receptors were aggregated; disaggregation of the receptors resulted in rapid dephosphorylation of p60. The phosphorylation of p60 was only detected after activation by IgE and antigen and not by stimulation with PMA and/or ionomycin. Phosphorylated p60 was associated entirely with the membrane fraction of the cells. A considerable fraction of Csk was associated with the membrane in both unstimulated and stimulated cells, this fraction did not change upon activation. p60 coprecipitated with Csk from both unstimulated and FcepsilonRI stimulated cells and was phosphorylated by the immunocomplex. Total kinase activity of Csk immunoprecipitates increased upon FcepsilonRI stimulation. p60 did not react with antibodies to a number of known signaling molecules, including the recently cloned, GAP-associated protein, p62dok. Our data demonstrate that Csk associates with a membrane-anchored protein complex that is directly involved in FcepsilonRI signal transduction. PMID- 9736342 TI - Ostial renal artery stent placement for atherosclerotic renal artery stenosis in patients with coronary artery disease. AB - To test the utility of endoprosthetic treatment for ostial renal artery stenosis, and to examine blood pressure and its treatment, serum creatinine, and restenosis rate, 44 ostial renal stent placements were performed in 30 patients with concomitant coronary artery disease, arterial hypertension, and the indication for angiotensin converting enzyme (ACE) therapy. There was a marked decrease in systolic and diastolic blood pressure (163+/-30 to 145+/-17 and 93+/-18 to 83+/ 10 mm Hg; P < 0.008) with a decrease in number of medication (3.2+/-0.9 to 2.8+/ 1.0; P = 0.005). In 5 out of 8 patients not receiving an ACE inhibitor, this drug could be added. Serum creatinine changed from 1.46+/-0.7 mg/dl to 1.39+/-0.58 mg/dl (P = ns). Three patients showed restenosis (12.5%). Ostial stenting lowers blood pressure, decreases antihypertensive drugs and increases medication flexibility. PMID- 9736343 TI - Open renal arteries are better than closed renal arteries. PMID- 9736344 TI - Simultaneous bilateral carotid stenting for restenosis after endarterectomy. AB - Carotid stenting may offer an alternative to surgery for recurrent carotid artery stenosis following endarterectomy. Five patients successfully underwent percutaneous simultaneous bilateral carotid artery stenting for restenosis after endarterectomy, without complications. Six months of clinical and imaging follow up showed preserved patency of all vessels treated, with no neurologic events in any of the patients. PMID- 9736346 TI - Can digital subtraction myocardial perfusion imaging studies offer information for postangioplasty lesion assessment? PMID- 9736345 TI - Comparison of coronary flow velocity and regional myocardial perfusion for functional evaluation of coronary artery disease in the setting of angioplasty. AB - Two essentially different methods for physiological evaluation of coronary artery disease were compared in the setting of angioplasty and related to quantitative coronary angiography. Forty-five patients, referred for percutaneous transluminal coronary angioplasty (PTCA), were examined by digital subtraction angiography (DSA) and by coronary flow velocity measurements distal to the target stenosis. Before PTCA, hyperemic mean transit time (HMTT) was correlated with % area stenosis r = 0.56*, coronary flow velocity reserve (CFVR) r = 0.58* and with CFVRN (CFVR normalized to a mean blood pressure of 100 mmHg) r = 0.68*. The correlation between CFVR and % area stenosis was r = 0.72* (*P < 0.001). After PTCA, all correlations between these measurements disappeared. HMTT and CFVR remained abnormal in 18% and 32 % of the patients, respectively. Pre-PTCA, distal coronary flow velocity measurements were reasonably well related to the assessment of regional myocardial perfusion. Flow velocity parameters, however, were better related to angiographic stenosis parameters. After PTCA, HMTT showed a more consistent improvement compared to CFVR. Flow velocity measurements appear to be more useful for the evaluation of local coronary stenoses, whereas the assessment of regional myocardial perfusion by DSA may be used for a more general evaluation of vessel territories. PMID- 9736347 TI - Interventional catheterization decreases plasma levels of atrial natriuretic peptide (ANP) in children with congenital heart defects. AB - Atrial natriuretic peptide (ANP) is one of the cardiac peptides implicated in volume and sodium homeostasis. We investigated the effect of interventional catheterization on plasma levels of ANP, aldosterone, and cortisol in 28 children with various congenital heart defects (CHD). Patients were divided by age into two groups: group A--infants and children over 3 months of age (n = 22), and group B--newborns (n = 6). These were compared to age-matched control groups. In group A, interventions included pulmonic valvotomy (n = 8), aortic valvotomy (n = 4), balloon angioplasty of native coarctation of the aorta (n = 3), balloon dilatation of the mitral valve (n = 1), and Rashkind double umbrella closure of patent ductus arteriosus (n = 6). Group B interventions included pulmonic valvotomy (n = 3), aortic valvotomy (n = 1), and balloon atrial septosomy (n = 2). In group A, mean ANP levels were markedly higher than in age-matched controls (125.2+/-15.8 vs. 24.6+/-4.6 pg/ml) (P <0.0001), and decreased immediately after intervention (75.6+/-11.4 pg/ml, P <0.02), and more markedly on follow-up (42.9+/ 5.0 pg/ml, P < 0.0001). In group B (newborns), mean basal plasma levels were high before and after intervention and were not different from age-matched controls (243+/-42.1 vs. 220.8+/-16.2 pg/ml). There was a significant decrease on follow up measurement (62.1+/-12.7 pg/ml, P < 0.005). In both groups, plasma cortisol levels increased significantly immediately following catheterization (P < 0.02), and normalized on follow-up. Basal aldosterone levels were normal in group A and high in Group B (9.9+/-3.8 vs. 167.6+/-16.9 ng/dl) (P < 0.001). It is suggested that plasma ANP levels are increased in children with CHD, without overt heart failure, and decrease significantly following successful intervention. In newborns with CHD, the physiological high ANP levels obscure the effect of the CHD. PMID- 9736348 TI - Balloon mitral valvuloplasty employing double transseptal puncture. AB - Patients with mechanical aortic valve prostheses occasionally develop subsequent mitral stenosis and present as candidates for further intervention. Repeat thoracotomy in such patients carries considerable operative risk, but the traditional alternative of balloon mitral valvuloplasty with transaortic intraventricular monitoring is not feasible because mechanical aortic prostheses cannot be safely crossed at catheterization. We have therefore developed a technique for performing the procedure via double transseptal puncture. We present the technique and our experience of its use in four patients with mechanical aortic prostheses presenting for balloon mitral valvuloplasty. PMID- 9736349 TI - Radiofrequency pulmonary valvotomy using a new 2-French catheter. AB - We report the usefulness of a new 2-French (F) electrode catheter for perforating the atretic pulmonary valve in patients with pulmonary atresia and intact ventricular septum, using radiofrequency energy. The new 2-F electrode catheter was used in three patients. The first patient, weighing 2.1 kg, with pulmonary atresia and an intact ventricular septum, underwent transcatheter valvotomy at the age of 18 days. Due to massive left ventricular volume overload, the patient required surgical ligation of the ductus arteriosus, but she was discharged uneventfully after that. The second patient, weighing 2.2 kg, had Ebstein's anomaly with pulmonary atresia and an intact ventricular septum. She underwent transcatheter valvotomy at the age of 30 days. Although she was weaned from prostaglandin E1 infusion, she died suddenly (presumed septicemic). A postmortem examination showed a split pulmonary valve. The third patient, weighing 2.3 kg, with pulmonary atresia and an intact ventricular septum, underwent transcatheter valvotomy at the age of 17 days. Prostaglandin E1 infusion was discontinued on the 3rd day after the transcatheter valvotomy and she was discharged uneventfully. PMID- 9736350 TI - Another new perforating toy for the pediatric interventionalist treating neonates with pulmonary atresia and intact ventricular septum...as long as you live outside of the United States! PMID- 9736351 TI - Catheterization of the pulmonary artery using a 3 French catheter in patients with congenital heart disease. AB - A 3 French (3F) coaxial polyurethane catheter was utilized for catheterization of the pulmonary artery in patients with complex congenital heart disease with concomitant stenosis or atresia of the pulmonary valve, with or without Blalock Taussig shunts. Mean aortic and atrial pressures measured with the 3F catheter were compared with those measured with 5-6F conventional catheters. The values measured with the 3F catheter were identical to those measured with 5-6F catheters. Mean pulmonary arterial pressure measured using the 3F catheter was significantly higher than that measured using the 5-6F catheters, but it was not significantly different from mean pulmonary venous wedge pressure. Of 43 patients, 5-6F conventional catheters could be inserted into the branch pulmonary artery in 22 patients, but the 3F catheter could be inserted into the pulmonary artery in all patients. These data suggest that the 3F catheter is useful for catheterization in patients with complex congenital heart disease associated with pulmonary atresia or severe pulmonary stenosis. PMID- 9736352 TI - Combined angioplasty and valvuloplasty for coexisting coarctation of the aorta and aortic stenosis in an infant. AB - The patient was a 1-month-old male infant. Echocardiography showed left ventricular hypertrophy with coarctation of the aorta (CoA) and aortic valve stenosis (AVS). Left ventricular systolic function was poor. At catheterization, the gradient of AVS was 25 mmHg and the annulus size of the aortic valve was 7 mm. There was a CoA measuring 2.1 mm in diameter. Balloon valvuloplasty of the CoA was then performed using the same balloon; the pressure gradient decreased to 0 mmHg, and CoA diameter decreased from 2.1 to 5.5 mm. PMID- 9736353 TI - Stent by stent crush: procedural outcome and angiographic follow-up. AB - Rewiring of an occluded and incompletely deployed coronary stent with a sharp and/or tortuous entrance angle may be difficult in the emergency setting. We report the case of a patient who experienced subacute stent thrombosis in the proximal right coronary artery. Incorrect rewiring resulted in stent crush after conventional balloon angioplasty at lesion site. In order to improve the angiographic result parallel stent placement was performed. Repeat angiography at 6 months demonstrated vessel patency without restenosis. PMID- 9736354 TI - Therapeutic repositioning of a Gianturco-Roubin II coronary stent after initial deployment. AB - Movement of coronary stents after deployment can produce complications. We report a case of stent migration that led to stent coverage of a distal dissection, obviating the need for placement of a second stent. In this case, stent movement was therapeutic. PMID- 9736355 TI - Complete left atrial obliteration due to localized tamponade after coronary artery perforation during PTCA. AB - Coronary artery perforation is a rare but important complication of percutaneous revascularization (PTCA). Clinical events following coronary perforation may include cardiac tamponade. After bypass graft operation (CABG), however, cardiac tamponade with subsequent hemodynamic instability is unusual due to the development of pericardial adhesions. We report an unusual case of localized tamponade after coronary artery perforation during PTCA in a patient with previous CABG. PMID- 9736356 TI - Emergency left main stenting in the management of postcoronary bypass graft surgery (CABG) ischemia. AB - Angioplasty has been used in the management of postcoronary bypass graft myocardial ischemia/infarction. A stent was successfully deployed in the left main artery in a patient with postcoronary bypass graft ischemia with hemodynamic instability. This case illustrates the potential use of emergency left main stenting in a selected patient with peri-operative ischemia, who was considered high risk for re-operation. PMID- 9736357 TI - Case of successful percutaneous stenting of an in situ gastroepiploic-coronary bypass graft. AB - The in situ or free gastroepiploic artery (GEA) is being used as an arterial conduit for coronary artery bypass surgery (CABG). The recent rapid improvements in stent manufacture, particularly their profile and flexibility, and related equipment, have helped reduce complications of coronary angioplasty. We describe one case of successful stenting of an in situ GEA-posterior descending artery graft. Stenting of an in situ GEA graft may avoid an incomplete result of angioplasty with possible restenosis or the need for multiple surgical revascularization. The use of stents in GEA grafts with 6 Fr soft guiding catheters can be encouraged. PMID- 9736358 TI - Hemodynamic rounds series II: hemodynamic evaluation of a stenotic bioprosthetic mitral valve. PMID- 9736359 TI - What is radiation dose? A tutorial module. PMID- 9736360 TI - Novel vascular sealing device for closure of percutaneous vascular access sites. AB - To investigate the hemostatic capabilities of a novel vascular sealing device consisting of a balloon catheter and procoagulant, vascular sheaths were placed percutaneously in the femoral arteries of dogs. The sealing device was evaluated using the balloon catheter alone in six femoral arteries and with the addition of a procoagulant, in 21 femoral arteries. The balloon catheter alone was successfully deployed in six of six femoral arteries achieving immediate hemostasis. In a second study in which the procoagulant was delivered following balloon placement, the sealing device was successfully deployed and hemostasis was achieved in 20 of 21 attempts (95%) despite removal of the balloon catheter. In a subset of fully anticoagulated animals, hemostasis was achieved in the sealing device-treated arteries at 6.5+/-3.4 minutes, but in none of the controls (P < 0.001). This novel vascular sealing device successfully achieves rapid hemostasis in normal and anticoagulated dogs following percutaneous vascular procedures. PMID- 9736362 TI - Bifuration coronary angioplasty using a new side branch accessible stent. AB - Two cases of successful angioplasty of bifurcational left anterior descending and diagonal lesions treated with a new side branch accessible Jomed stent in the LAD are discussed. A balloon dilatation of the side branch using kissing balloon technique in the first case and stenting the side branch followed by kissing balloon dilatation in the second case are described. PMID- 9736361 TI - Percutaneous vascular closure: plugs, stitches, and glue. PMID- 9736363 TI - Impact of Doppler guidewire size and flow rates on intravascular velocity profiles. AB - Coronary blood flow velocity measurements by conventional intravascular catheter based Doppler devices are well known to be affected by catheter size. Moreover, it is of clinical importance that the assessment of maximum vasodilator capacity, i.e., the coronary reserve, might be considerably affected by a shape change of the velocity profile under hyperemia. Therefore, the present in vitro study aimed to assess the impact of a small-size Doppler guidewire on the velocity profiles interrogated in tubes with diameters corresponding to the epicardial coronary arteries at clinically relevant flow rates. A 0.014" guidewire was inserted into four serially connected silicone tubes of known diameter, which were perfused with discarded human whole blood by means of a roller pump. In order to determine the effect of the Doppler guidewire on the velocity profile antegrade and retrograde perfusion were carried out in each vessel segment. Vm, the true mean velocity, was calculated from the time collected flow divided by the corresponding vessel cross-sectional area. Average peak velocity (APV) measurements were obtained by pulling back the Doppler device across each of the four vessel segments with given flow rates ranging from 1.14-5.88 ml/s in antegrade and retrograde direction. The shape factor of the recorded velocity profile (fp) is defined as vm/APV and is generally assumed to be close to 0.5. Antegrade perfusion: APV = 1.63 vm + 5.35 (R2 = 0.98); fp = 0.001 vm + 0.51. Retrograde perfusion: APV = 1.71 vm + 3.33 (R2 = 0.98); fp = 0.001 vm + 0.52. Due to the constant relationship between APV and vm, velocity profiles within vessels of epicardial coronary artery size are not substantially disturbed by the presence of the Doppler guidewire. The slight, but significant increase of experimental fp with increasing flow is at variance with the theoretically expected fp values. PMID- 9736364 TI - Re Antonellis and Patsilinakos. Catheterization of coronary arteries in the presence of tortuous iliac arteries. PMID- 9736365 TI - Percutaneous balloon mitral valvotomy as a bridge to elective mitral valve replacement. PMID- 9736366 TI - Effect of allogenic freeze-dried demineralized bone matrix on guided tissue regeneration in dogs. AB - This randomized, split-mouth study was designed to evaluate the adjunctive effect of allogenic, freeze-dried, demineralized bone matrix (DBM) to guided tissue regeneration (GTR). Contralateral fenestration defects (6 x 4 mm) were created 6 mm apical to the buccal alveolar crest on maxillary canine teeth in 6 beagle dogs. DBM was implanted into one randomly selected fenestration defect. Expanded polytetrafluoroethylene (ePTFE) membranes were used to provide bilateral GTR. Tissue blocks including defects with overlying membranes and soft tissues were harvested following a four-week healing interval and prepared for histometric analysis. Differences between GTR+DBM and GTR defects were evaluated using a paired t-test (N = 6). DBM was discernible in all GTR+DBM defects with limited, if any, evidence of bone metabolic activity. Rather, the DBM particles appeared solidified within a dense connective tissue matrix, often in close contact to the instrumented root. There were no statistically significant differences between the GTR+DBM versus the GTR condition for any histometric parameter examined. Fenestration defect height averaged 3.7+/-0.3 and 3.9+/-0.3 mm, total bone regeneration 0.8+/-0.6 and 1.5+/-0.8 mm, and total cementum regeneration 2.0+/ 1.3 and 1.6+/-1.7 mm for GTR+DBM and GTR defects, respectively. The histologic and histometric observations, in concert, suggest that allogenic freeze-dried DBM has no adjunctive effect to GTR in periodontal fenestration defects over a four week healing interval. The critical findings were 1) the DBM particles remained, embedded in dense connective tissue without evidence of bone metabolic activity; and 2) limited and similar amounts of bone and cementum regeneration were observed for both the GTR+DBM and GTR defects. PMID- 9736367 TI - In vitro studies on the effect of cleaning methods on different implant surfaces. AB - The effect of specific cleaning procedures was examined on the surfaces of 3 implant types with different coatings and shapes (plasma sprayed [PS]; hydroxyapatite coated [HA] implants; and smooth titanium surface screws) using a scanning electron microscope. Each implant was treated for 60 seconds per instrument with one of 6 different hygiene measures: plastic curet, metal curet, diamond polishing device, ultrasonic scaler, air-powder-water spray with sodium hydrocarbonate solution, and chlorhexidine 0.1% solution rinse. The air-powder abrasive system, chlorhexidine rinse, and curettage with a plastic instrument caused little or no surface damage in all but the hydroxyapatite-coated fixtures. Therefore, these 3 methods were tested to determine their cleaning efficacy in a second clinical study, which did not include the HA-coated fixture. Two implants were placed on the facial aspects of both upper molar regions using individual acrylic plates. Thus, 2 fixtures on each side were examined in each patient. The examination revealed that only the sodium hydrocarbonate spray yielded a clean fixture without damage to the implant surface. In a third stage, which imitated the clinical procedure of the second approach, the cell growth of mouse fibroblasts on implant surfaces was examined after cleaning the surface with plastic scaler and the air-abrasive system, which represents the least damaging and most effective methods. In contrast to the implant surfaces treated with plastic scalers, mostly vital cells were found on implants sprayed with the air abrasive system. PMID- 9736368 TI - The relationship between concentrations of proinflammatory cytokines within gingiva and the adjacent sulcular depth. AB - The objective of this study was to determine and compare concentrations and ratios of 3 proinflammatory cytokines, interleukin (IL) IL-1beta, IL-6, and IL-8 within gingival tissue biopsies adjacent to < or = 3, 4 to 6, or >6 mm sulci. All gingiva adjacent to > or = 4 mm sulci had clinical evidence of active inflammation. Factorial analysis of variance suggested significant effects of sulcus depth on the type and concentration of the three cytokines in the adjacent gingiva (P < 0.001). IL-8 concentrations were highest in gingiva adjacent to < or = 3 and lowest adjacent to >6 mm sulci (P < 0.001). In contrast, IL-6 concentrations were lowest in gingiva adjacent to < or = 6 mm and highest adjacent to >6 mm sites. IL-1beta concentrations were highest in gingiva adjacent to >6 mm and lowest adjacent to 4 to 6 mm sites; they were also higher adjacent to < or = 3 mm than adjacent to 4 to 6 mm sites (P < 0.01). Multiple regression analysis suggested that sulcular depth, type of cytokine, and cytokine concentration were significantly correlated (P < 0.001). Ratios of gingival cytokines changed with increased sulcular depth. In gingiva adjacent to < or = 6 mm sites, IL-8 was the most and IL-6 the least prevalent. In gingiva adjacent to > or = 6 mm sites, IL-8 was the least and IL-1-beta the most prevalent. The data suggest that the characteristics of the gingival cytokine network are affected by adjacent sulcular depth. These data could be used to design adjunct diagnostic tests for progression of periodontal diseases. PMID- 9736369 TI - The effect of different diagnostic thresholds on incidence of disease progression. AB - Detection of periodontal disease progression occurs when a predetermined threshold of attachment loss is exceeded during longitudinal monitoring. The incidence of disease progression in a population may be dependent on the method and threshold utilized to identify significant changes in attachment level measurements. The aim of this study was to investigate the effect of utilizing different methods and thresholds on the incidence of disease progression in an untreated periodontitis population. The relationship between baseline clinical parameters and disease progression was also examined. A total of 411 interproximal sites in 46 individuals were monitored monthly over a 6-month period. Disease progression was determined by the cumulative sum (CUSUM) method and by the absolute change in relative attachment level between months 0 and 6 utilizing 3 different thresholds for attachment level change (0.58 mm, 1.16 mm, and 1.74 mm) based upon examiner repeatability using an automated probe. Utilizing the CUSUM method, 49 of 411 sites (11.9%) demonstrated attachment loss over the 6-month observation period. When attachment level changes > or = 0.58 mm, > or = 1.16 mm, and > or = 1.74 mm were used to identify disease progression, the percentage of sites exhibiting deterioration were 19.5%, 8.8%, and 2.9%, respectively. These results demonstrate that the apparent incidence of disease progression was dependent on the method and threshold utilized to detect progressive sites. When utilizing the CUSUM and 0.58 mm thresholds a significant (P < 0.05), but weak relationship (r = -0.26) was observed between baseline relative attachment level measurements and sites exhibiting disease progression. This finding suggests that sites with significant but relatively less attachment loss may be more likely to experience further breakdown compared to sites with a history of greater periodontal destruction. PMID- 9736370 TI - Calprotectin and lactoferrin levels in the gingival crevicular fluid of children. AB - The purpose of this study was to determine the levels of calprotectin and lactoferrin, 2 microbiostatic proteins, in the gingival crevicular fluid (GCF) of normal children. The children represented a population, primarily underprivileged, seeking care at a regional dental health care center. GCF was collected from Ramfjord teeth (or their deciduous equivalent). GCF volume was quantified by conductance. Calprotectin and lactoferrin levels were quantified by sandwich ELISA, and found to have a mean value of 70.8+/-94.2 microg/mL and 68.2+/-108.7 microg/mL, respectively. The levels of calprotectin and lactoferrin varied directly with one another and inversely with the amount of fluid obtained in a 20-second sampling period. The mean levels were at or above the minimum inhibitory concentration determined in vitro. PMID- 9736371 TI - Heat generation during ultrasonic instrumentation of dentin as affected by different irrigation methods. AB - Heat is produced by magnetostrictive ultrasonic scalers which may cause injury to pulpal and periodontal tissues. Water coolant flows around the instrument stack and is directed at the instrument tip to reduce the generation of heat. Sound surgical practice requires the use of a sterile coolant for ultrasonic scaling during surgery. Intermittent bulb irrigation is one way to deliver sterile coolant when using ultrasonic scalers not equipped with a dedicated sterile water reservoir. The purpose of this study was to compare the temperature rise in dentin during ultrasonic scaling using either ultrasonic handpiece irrigation or intermittent bulb irrigation. Twenty dentin/cementum root slabs were prepared for each thickness of 0.5, 1.5, and 2.5 mm. A 3.0 mm x 1.5 mm field was outlined on each slab to indicate the area of intended instrumentation. Each slab was mounted such that a thermocouple placed in contact with dentin opposite the area of instrumentation was shielded from irrigation. Twenty samples of each thickness were ultrasonically scaled during which dentin temperature was recorded every 5 seconds over a 30-second period. All 60 slabs were first treated with dental unit ultrasonic handpiece water irrigation, followed by no irrigation, and finally by bulb irrigation with sterile saline. Repeated measures analysis of variance indicated that there were differences among the three treatment groups for temperature change over the course of the study (P < 0.001). Dentin temperature increased with both decreasing slab thickness and with increasing duration of instrumentation. However, only scaling without irrigation produced a rise in dentin temperature from baseline to a level reported as deleterious to pulpal and periodontal tissues. Bulb syringe irrigation delivered as a continuous drip and ultrasonic unit water spray minimized heat generation to physiologically tolerable levels. Intermittent bulb irrigation appears to be a satisfactory alternative to use of ultrasonic scaler water spray for cooling during ultrasonic scaling at surgery. PMID- 9736372 TI - Nature and attachment of cementum formed under guided conditions in human teeth. An electron microscopic study. AB - In an attempt at characterizing the nature and attachment of cementum formed under conditions of guided tissue regeneration (GTR) in humans, front teeth from 4 patients aged 42 to 72 years were examined at the electron microscopic level. All teeth were affected by complex periodontitis associated with advanced loss of periodontal support. Roots were surgically planed and notched, but not chemically conditioned. Either the mesial or distal surface of each tooth represented the experimental site and was covered with a biodegradable polyglactin 910 barrier, while the opposite approximal surface served as control. Following 3 months of healing, teeth were removed together with surrounding periodontal tissues including some alveolar bone. These blocks were fixed histologically, decalcified, embedded in epoxy, and sectioned for examination in the scanning (backscatter mode) and transmission electron microscope. Both experimental and control sites disclosed 2 types of regenerative cementum that seemed to be formed by cells resembling cementoblasts. The first type was characterized by a thin fringe of collagen fibrils which were arranged perpendicular to the root surface and appeared mineralized in a zone extending about 1 to 3 microm from the dentin. The second type occurred as thick patches which revealed scattered cementocytes and sheets of collagen fibrils oriented mainly parallel to the root surface, running both circularly and axially. In both situations, a continuous, thin, electrondense layer was interposed between newly formed cementum and preexisting radicular hard tissues. Interdigitation of collagen fibrils from cementum and dentin, such as observed along the natural cemento-dentinal junction, did not occur. Thus, regenerative cementum laid down in humans under guided conditions on previously diseased and planed, but not otherwise treated root surfaces shares some morphologic features with cementum formed during spontaneous repair of root resorptions. However, unlike in the course of such repair, a fibrous attachment of new cementum resembling the natural cemento-dentinal junction does not seem to be regenerated under guided conditions. PMID- 9736373 TI - Increased presence of interleukin-6 (IL-6) and IL-8 secreting fibroblast subpopulations in adult periodontitis. AB - Periodontitis is a chronic inflammation of the supporting structures of the dentition which constitutes one of the most common causes of adult tooth loss. While certain microorganisms have been associated with the onset of the disease process, the exact pathogenetic mechanisms underlying periodontal destruction are still poorly understood. We have tested the hypothesis that gingival fibroblasts from diseased sites contribute to pathogenesis by possessing a secretory phenotype characterized by an exuberant secretion of inflammatory mediators and cytokines. Of the cytokines and mediators tested, fibroblast IL-1beta and prostaglandin E2 (PGE2) secretion was not different between health and disease. However, we have shown that fibroblasts from periodontal lesions produce in vitro greater amounts of IL-6 and IL-8 constitutively than healthy controls. When fibroblasts were stimulated with a panel of endogenous or exogenous response modifiers, the magnitude of cytokine and mediator stimulation above constitutive levels did not differ between health and disease. A strong positive correlation was identified between IL-6 or IL-8 constitutive secretion levels in vitro and the in situ expression of these cytokines within the connective tissues from where these cells originated, indicating that the in vitro phenotype mirrors their in vivo function. Furthermore, we present evidence which indicates that increased cytokine secretion by fibroblasts in disease is due to an elevated proportion of subpopulations with higher cytokine secretory capacity. Finally, we demonstrated that cultures from diseased sites are composed of cells with higher levels of constitutive CD40 expression, which may contribute to the increased IL 6 and IL-8 secretory phenotype. PMID- 9736374 TI - Maxillary sinus augmentation in the non-human primate: a comparative radiographic and histologic study between recombinant human osteogenic protein-1 and natural bone mineral. AB - The posterior maxilla has traditionally been one of the most difficult areas to successfully place dental implants due to poor bone quality and close approximation to the maxillary sinus. Sinus augmentation procedures have become a viable means of assuring adequate bone for the placement of dental implants in this area. However, with the techniques currently employed, a considerable variation in the quality of bone attained with the sinus augmentation procedure exists. The purpose of this in vivo study was to evaluate the healing response and bone formation stimulated by 3 doses of recombinant human osteogenic protein 1 (rhOP-1), 0.25, 0.6, and 2.5 mg OP-1 per gram of collagen matrix; natural bone mineral; or collagen matrix alone (control) placed in the maxillary sinus of adult chimpanzees. Results were assessed using clinical, histologic, and radiographic techniques. Radiographic analysis of the computed tomography scans taken at 1 week, and 2.5, 4.5, and 6.5 months revealed a more rapid mineralization with the 2.5 mg OP-1/g collagen matrix and natural bone mineral treatment groups. The incremental bone mineral density (BMD) increase for these 2 treatments from 1 week to 2.5 months was over 2.5 times the increase found with the collagen matrix alone; these 2 treatments also had a higher BMD at the most superior slices evaluated when compared to the other 3 groups. Biopsy specimens were taken at 3.5, 5.5, and 7.5 months and for all 5 treatment groups bone formation was observed at all time points in the majority of the specimens. At 7.5 months the 2.5 and 0.6 mg OP-1/g collagen matrix treatment groups had an increase in the percent bone area when compared to the matrix alone control. In conclusion, these results demonstrate that sinus augmentation with natural bone mineral or 2.5 mg OP-1/g collagen matrix induce comparable radiographic and histologic evidence of bone formation and that both of these treatments performed superior to the control group of collagen matrix alone based upon all methods of evaluation. PMID- 9736375 TI - A prospective clinical trial of endosseous screw-shaped implants placed at the time of tooth extraction without augmentation. AB - This prospective clinical trial evaluated 134 implants in 81 patients. The implants were placed at the time of tooth extraction and were not augmented with barrier membranes or graft materials. The implants were placed into good jaw bone anatomy and quality and were restored by dentists familiar with the implant system. Forty-seven implants were followed between 4 to 5 years with a cumulative success rate of 93.3%. Marginal bone levels were measured for 61 patients with 108 implants. The average mesial-distal measurements for maxillary implants at abutment connection were 1.02 mm (SD+/-0.59) and 1.36 mm (SD+/-0.78) at an average of 32 months follow-up. These differences were not significant. The average mandibular mesial-distal measurements at abutment connection were 1.05 mm (SD+/-0.92) and 1.54 mm (SD+/-0.91) at follow-up. These differences were statistically significant (P = 0.0027). Removal of one patient (5 implants) with advanced marginal bone loss from the data provided a marginal bone level of 1.20 mm (SD+/-0.94) at abutment connection and 1.30 mm (SD+/-0.87) at follow-up. These differences were not significant. The results of this study indicate that implants placed at the time of extraction without augmentation or grafting have excellent long-term cumulative success rates. PMID- 9736377 TI - Carious lesions in the heroin addicted patient. A case report. AB - Heroin abuse is destroying the health of many individuals in our society. Much of the information that has appeared in the literature has concerned itself with the general health of addicts; the oral health status of these individuals has not been studied to the same extent. This report will present and discuss the management of a case involving the effects of long-term heroin abuse on the dentition. PMID- 9736376 TI - The effects of guided bone regeneration and grafting on implants placed into immediate extraction sockets. An experimental study in dogs. AB - Guided bone regeneration (gbr) for the treatment of insufficient bone volume around implants can be performed using membranes with or without grafting materials (i.e., autogenous, allogenous, xenogenous, or alloplastic grafts). A possible way to evaluate the quality of implant osseointegration is the torque necessary to remove implants from their bony housing. The aim of this study was to compare the torques necessary to remove dental implants from implant beds reconstructed with different bone substitutes and GBR or GBR alone in 6 adult mongrel dogs. All mandibular premolars were extracted and 3 extraction sockets on each side were enlarged using a trephine bur. A 13 mm titanium screw-type dental implant (3.75 mm diameter) was placed in each enlarged extraction socket so that only the apical 3 to 4 mm were engaged in bone. The 3 defects were then randomly treated with either 1) canine demineralized freeze-dried bone allograft (DFDBA) plus GBR using an expanded polytetrafluoroethylene membrane (DFDBA+GTAM); 2) bioabsorbable hydroxyapatite and GBR (HA+GTAM); or 3) GBR (GTAM alone). After 6 months, the torque to remove the implants was measured in 4 animals and analyzed using ANOVA. There were no statistically significant differences between the 3 groups (GTAM alone: 46.37+/-16.41 Ncm; HA+GTAM: 46.00+/-16.59 Ncm; DFDBA+ GTAM: 52.15+/-29.24 Ncm). In addition, the influence of early removal of barriers on the torque values was evaluated with the t-test. Comparing exposed versus retained membranes by treatment modality, the only statistically significant difference was found in the DFDBA+GTAM group. When the torque values of all implants with exposed and retrieved membranes were compared to all those with retained membranes a significant difference could be detected. Histologic sections were prepared from the 2 dogs not included in the removal torque testing. In the histometric analysis the GTAM alone group showed a mean mineralized bone-to-implant-contact of 27.1%, the DFDBA+GTAM group of 34.6%, and the HA+GTAM of 39.3%. The mineralized bone-to-implant-contact of the HA+GTAM group was significantly higher than that of the GTAM alone group. In addition, the mineralized bone-to-implant-contact was divided into an apical and coronal part using the apical seventh thread as the dividing landmark. In the apical region, there was no significant difference between the groups regarding mineralized bone-to-implant-contact. In the coronal part the mineralized bone-to implant-contact of the GTAM alone group was significantly lower compared to the other 2 groups. Within the limits of this investigation, it can be concluded that the type of grafting material will not influence torque removal values, but that early membrane exposure and removal will negatively influence the torque measurements. The combination of GBR with a bone substitute increased the mineralized bone-to-implant contact. PMID- 9736378 TI - A new approach in restorative treatment of external root resorption. A case report. AB - This case report describes the treatment of an external root resorption with extensive loss of tooth structure and bone at the labial surface of an upper left central incisor. The area of bone loss and root resorption was surgically exposed and an impression was taken using curing silicone. An individual ceramic insert was fabricated, allowing endodontic retreatment through an artificial root canal. The insert was incorporated using a dentin bonding system and a dual curing luting composite. Following endodontic retreatment and internal bleaching, a ceramic veneer was bonded to the tooth to obtain good esthetics and to improve stability. Twenty months after surgical treatment no further root resorption could be detected radiographically. A shallow residual pocket but no bleeding on probing was found. PMID- 9736379 TI - The treatment of class III maxillary furcations using a resin-ionomer. A case report. AB - This case report uses a resin-ionomer restoration as a barrier in the treatment of a Class III furcation defect. There was a reduction in tooth mobility and plaque count, no bleeding on probing, and a decrease in probing depth with the use of the resin ionomer. The study offers another treatment option in the treatment of a seemingly hopeless maxillary molar. PMID- 9736380 TI - Current understanding of the role of microscopic monitoring, baking soda, and hydrogen peroxide in the treatment of periodontal disease. Committee on Research, Science and Therapy. The American Academy of Periodontology. AB - The Keyes technique came to national attention following a reference in The New York Times in the late 1970s. Several lay press articles and discussions on national television served to further focus the interest of patients, general dentists, and periodontists on this potential approach to periodontal therapy. Early evaluations of the data on the technique resulted in an Academy position paper in 1981. Recognizing that there was a lack of well-controlled studies on the technique led to extensive research efforts supported by the National Institute of Dental Research. The results of those efforts have provided substantial new information that serves as the basis of the present position paper. Although this technique is no longer widely used in the United States, some patients and dentists may have not had the benefit of the new data. PMID- 9736381 TI - Evaluation of a hand-held electrical conductivity meter for detection of subclinical mastitis in cattle. AB - OBJECTIVE: To assess, under field conditions, whether a hand-held electrical conductivity (EC) meter could be used to detect subclinical mastitis caused by pathogens most commonly associated with mastitis in dairy cows. ANIMALS: 425 lactating cows on 15 dairies in Costa Rica. PROCEDURE: Immediately prior to milking, milk samples from each quarter were tested, using a hand-held EC meter. A milk sample from the quarter with the highest score was submitted for bacteriologic culture. Results of bacteriologic culture were compared with highest absolute EC score for each cow and with differential EC score (ie, difference between the highest and lowest absolute EC scores for the 4 quarters of each cow). RESULTS: Absolute EC score for cows with subclinical mastitis was significantly higher than that for cows without subclinical mastitis, and absolute EC score was significantly associated with detection of subclinical mastitis. If absolute EC score > or = 7 was considered indicative of subclinical mastitis, sensitivity was 0.43, specificity was 0.83, predictive value of a positive result was 0.39, and predictive value of a negative result was 0.85. Differential EC score for cows with mastitis was significantly higher than that for cows without subclinical mastitis. If differential EC score > or = 2 was considered indicative of subclinical mastitis, sensitivity was 0.53, specificity was 0.77, predictive value of a positive result was 0.37, and predictive value of a negative result was 0.87. CLINICAL IMPLICATIONS: A hand-held EC meter may be used to screen cows for subclinical mastitis. PMID- 9736382 TI - Molecular diagnostic tests for ascertainment of genotype at the mucopolysaccharidosis type VII locus in dogs. AB - OBJECTIVE: To develop a molecular diagnostic test to ascertain genotype of the mucopolysaccharidosis type VII (MPS VII) locus. SAMPLE POPULATION: Blood samples from 45 mixed-breed (German Shepherd Dog X Beagle) dogs that were phenotypically normal or affected with MPSVII. PROCEDURE: The canine beta-glucuronidase gene (exon 3) was amplified by polymerase chain reaction (PCR), using 2 pairs of primers to determine the genotype of the MPSVII locus by 2 independent methods. For the first method, PCR products were used for heteroduplex analysis, using conformation-sensitive gel electrophoresis. In the second method, an allele specific restriction site was created by use of a mismatch primer in PCR. The amplified DNA fragment was digested with a restriction enzyme (Eag I) to enable identification of the wild-type and mutant alleles. RESULTS: Conformation sensitive gel electrophoresis resulted in a single DNA band representing homoduplex when the sample contained a wild-type or MPS VII allele, but 2 bands representing hetero- and homoduplexes when both alleles were in the sample. Restriction digestion of the DNA fragment obtained by use of a mismatch primer was cleaved only when the template was a wild-type allele. Thus, samples from phenotypically normal carrier dogs that contained wild-type and MPS VII alleles were partially digested by the enzyme. CONCLUSIONS: The diagnostic test used 2 strategies for independently ascertaining the wild-type or mutant MPS VII alleles in dogs. Thus, test results can distinguish phenotypically normal MPS VII-carrier dogs from homozygous normal dogs. PMID- 9736383 TI - Detection of milk antibiotic residues by use of screening tests and liquid chromatography after intramammary administration of amoxicillin or penicillin G in cows with clinical mastitis. AB - OBJECTIVE: To compare results of 6 commercially available milk antimicrobial screening tests with results of liquid chromatography (LC) when testing milk samples from individual cows treated for mild clinical mastitis by intramammary (IMM) infusion with amoxicillin or penicillin G. ANIMALS: 6 cows with noninduced clinical mastitis: 3 treated by IMM infusion with amoxicillin and 3 treated by IMM infusion with penicillin G. PROCEDURE: Composite milk samples were collected before, during, and after treatment. Samples were assayed by use of the screening tests and their results and those of LC were compared. The LC results were assumed to represent the true result. RESULTS: Results of screening tests compared well with results of LC, with agreement of 94%. Positive screening test results for samples containing drug values below the established tolerance or safe level, as evaluated by LC, were obtained from 2 cows in which abnormal milk, as well as marked increases in composite milk somatic cell count, were observed. With the exception of 1 test in 1 cow, all screening tests had negative results at the end of the labeled milk-withholding time. CONCLUSIONS AND CLINICAL IMPLICATIONS: On the basis of results of the limited sample reported, the screening tests appeared to provide good agreement overall, compared with LC results, when testing milk of individual cows treated by IMM infusion with amoxicillin or penicillin G. Positive screening test results for milk samples containing amoxicillin or penicillin G at values below the established tolerance or safe level, as evaluated by LC, may occasionally be obtained. PMID- 9736384 TI - Development of a simple enzyme immunoassay for blood haptoglobin concentration in cattle and its application in improving food safety. AB - OBJECTIVE: To verify the role of haptoglobin, a major acute-phase reactant protein in cattle, as a marker to identify health/disease status in cattle and further assess its potential in improving food safety. SAMPLE POPULATION: Serum samples from various cattle groups: clinically normal cattle comprising steers (n = 157) and culled dairy cows (n = 92) before death (antemortem [AM]); retained carcasses (n = 57) railed off the line during postmortem (PM) inspection; and apparently AM normal culled dairy cows (n = 57). PROCEDURE: Efficacy of the simplified monoclonal antibody-based enzyme immunoassay was established by comparing results of haptoglobin tests performed independently on aliquots of serum samples by 3 laboratories. RESULTS: Haptoglobin concentration was significantly (P< or = 0.0001) different between the PM retained carcass group (n = 57) and the AM steer (n = 157) and culled dairy cow (n = 92) groups. In addition, haptoglobin concentration in AM steers (n = 157) and culled dairy cows (n = 92) was significantly (P < or = 0.0012) different, possibly reflecting a higher percentage of underlying pathologic or inflammatory conditions in animals of the latter group. Evaluation in 3 laboratories of sera from a group of culled dairy cows (n = 57), each laboratory performing a different test procedure, indicated that correlation of haptoglobin concentrations was good between the reported test procedure and the unmodified test and the classical hemoglobin binding assay that measures peroxidase activity. CONCLUSION: Haptoglobin determination is effective in identifying diseased and healthy cattle. It may be a potentially important tool for application at the farm and slaughterhouse as an aid in improving food safety. PMID- 9736385 TI - Comparison of radiographic and necropsy findings of lung lesions in calves after challenge exposure with Pasteurella multocida. AB - OBJECTIVES: To test suitability of radiographic evaluation of lung lesions as a substitute for lung lesion scores derived by examination at necropsy in challenge exposure models of bovine pneumonia. ANIMALS: 10 calves selected by body weight from 20 multiple-source male Holstein calves approximately 1 to 2 months old enrolled in a Pasteurella multocida challenge-exposure study. PROCEDURE: Calves were paired on the basis of weight and randomly assigned within pairs to vaccine or control (saline solution) group. By use of deep tracheal cannulation, calves were challenge exposed with a culture of virulent P multocida, observed for 10 days, euthanatized, and necropsied, and the lungs were scored for pneumonic lesions. Radiographic views of the lung fields of the calves were taken before challenge exposure and before necropsy and were evaluated for alveolar disease by a veterinary radiologist. Lung lesion scores were compared with radiographic evaluations. RESULTS: There was a strong and significant correlation (R2 = 0.91, P < 0.001) between results of the evaluation of postchallenge-exposure radiographs and necropsy results. There also was also strong and significant correlation (R2 = 0.90, P < 0.001) between evaluation of the prechallenge exposure radiographs and necropsy results. CONCLUSIONS: Radiographic evaluation of lung lesions correlates well with lung lesions found at necropsy. The findings emphasize the need for caution in interpreting the results of challenge-exposure studies of bovine respiratory tract disease in which small numbers of calves are studied. PMID- 9736386 TI - Time course of gastrointestinal tract permeability to chromium 51-labeled ethylenediaminetetraacetate in healthy dogs. AB - OBJECTIVES: To establish values for gastrointestinal tract permeation by chromium 51-labeled ethylenediaminetetraacetate (51Cr-labeled EDTA) in healthy adult dogs, and to evaluate the time course for 51Cr-labeled EDTA absorption over a 24-hour period after its administration, in an effort to define a shorter, more practical collection method. ANIMALS: 6 healthy adult mixed-breed dogs. PROCEDURE: After an 18-hour nonfeeding period, each dog was given a solution containing 50 microCi of 51Cr-labeled EDTA in deionized water (10 ml/kg of body weight) by stomach tube. Complete urine collection was done at 2, 4, 6, and 24 hours after 51Cr-labeled EDTA administration. Five-milliliter samples of urine were counted for 15 minutes in a gamma counter, and radioactivity in urine was expressed as a percentage of the orally administered dose. RESULTS: Median (range) 24-hour urinary recovery of 51Cr-labeled EDTA after 24 hours was 15.1 (12.7 to 20.3)%. Urine collected at 2, 4, and 6 hours contained 1.0 (0.2 to 3.5)%, 6.5 (2.2 to 8.7)%, and 10.0 (8.1 to 11.7)% of the administered 51Cr-labeled EDTA, respectively. Urine passed during the first 6 hours contained, on average, 67 (54 to 77)% of the total 24-hour urine recovery. CONCLUSIONS: 6-hour urinary recovery of 51Cr-EDTA provides a potential alternative to 24-hour recovery. This shorter collection period may more specifically reflect small intestinal permeability. PMID- 9736387 TI - Evaluation of periparturient dairy cows and contact surfaces as a reservoir of Cryptosporidium parvum for calfhood infection. AB - OBJECTIVE: To determine whether periparturient cows or contact surfaces to which newborn calves are exposed are reservoirs of Cryptosporidium parvum oocysts. ANIMALS: Periparturient cows and their calves. PROCEDURE: Using direct fluorescent antibody (DFA) and acid-fast (AF) assays, fecal samples taken before and after calving from periparturient cows were tested for C parvum oocysts. Fecal samples from calves were collected every other day from age 7 to 21 days and were tested by use of the AF assay. Topsoil from close-up and maternity pens and scrapings from wooden walls and floors of calf hutches were tested for C parvum oocysts by use of DFA assay. RESULTS: None of the 384 fecal samples obtained 1 to 21 days before or after calving or on the day of calving from 154 periparturient cows contained detectable C parvum oocysts. Despite this lack of detectable periparturient shedding, the period prevalence of calfhood infection was 92% (123/134) from age 7 to 21 days. Soil samples from the close-up and maternity pens where newborn calves spend the first 12 hours of life also were negative for C parvum oocysts. Wood scrap ings from the outer 2 mm of the walls and floors of empty and cleaned calf hutches that were ready to receive calves were C parvum oocyst-positive. CONCLUSIONS: Conditional on sensitivity of DFA, periparturient cows did not appear to shed detectable C parvum oocysts. In contrast, the floors and walls of wooden calf hutches contained detectable C parvum oocysts on the surface. PMID- 9736388 TI - Role of horn flies (Haematobia irritans) in Staphylococcus aureus-induced mastitis in dairy heifers. AB - OBJECTIVE: To determine whether Staphylococcus aureus can colonize in horn flies and whether colonization is sufficiently persistent for transmission of the organism to cows by flies. ANIMALS: 2 Jersey heifers exposed to infected horn flies. PROCEDURE: Staphylococcus aureus was allowed to colonize in horn flies, and duration of colonization was determined. Flies with colonized S aureus were allowed to feed on teats of uninfected heifers to determine whether intramammary infection could be transmitted from fly to heifer. Scab material from naturally infected heifers was submitted for bacteriologic culture to determine whether S aureus was present and whether scabs could serve as a possible source of S aureus for flies. RESULTS: Staphylococcus aureus colonized in horn flies and remained for up to 96 hours after exposure. Exposure of teats of uninfected heifers to horn flies colonized with S aureus resulted in intramammary infection in 3 of 4 exposed teats. Culture of scab material from teats of naturally infected heifers revealed high concentration of S aureus (> 107 colony-forming units/mg), and flies without previously colonized S aureus were allowed to feed on scabs; S aureus colonized in them just as readily as it did in flies that had fed on experimentally infected blood. CONCLUSIONS: Horn flies are capable of transmitting S aureus-induced intramammary infection to heifers, and scabs on teats are a potential source of S aureus. Fly control on dairy cows in herds with known S aureus problems is recommended as a method to help prevent these infections. PMID- 9736389 TI - Characterization of Listeria monocytogenes isolated from channel catfish (Ictalurus punctatus). AB - OBJECTIVE: To characterize Listeria monocytogenes from tissues of channel catfish for their ability to cause hemolysis and grow intracellularly in mouse macrophages. SAMPLES: 15 isolates from processed fillets and 15 isolates from the brain, spleen, and kidneys. PROCEDURE: Serotype and hemolytic activity of L monocytogenes isolates were evaluated, using plate agglutination and CAMP tests, respectively. Invasiveness of L monocytogenes was determined by inoculating each strain or isolate on J774A.1 macrophage cells. Infected cells were incubated for 0 or 3 hours and lysed; then 100 gli of the lysate was plated onto a brain heart infusion agar plate. Colony counts for each strain or isolate were analyzed statistically. RESULTS: Of 30 isolates, 19 were serotype 1 and 11 were serotype 4. Mouse J774A.1 macrophages were inoculated with catfish isolates, a wild-type (EGD) or a nonhemolytic strain of L monocytogenes. Seventy-three percent (11/15) of isolates originating from catfish organs and 100% (15/15) of isolates originating from fillets were not significantly different from the wild-type EGD strain. The nonhemolytic L monocytogenes strain used as a negative control failed to replicate. Intracellular growth of all L monocytogenes isolates decreased after an additional 3-hour incubation period with medium containing 50 [microg/ml of gentamicin. CONCLUSIONS: Similar to the wild-type EGD strain, most channel catfish L monocytogenes isolates were hemolytic, serotype 1 or 4, and were invasive for mouse J774A.1 macrophages. CLINICAL RELEVANCE: monocytogenes growth in mouse macrophages may serve as an in vitro model for determining virulence of isolates from food products or environments. PMID- 9736390 TI - Molecular basis of variation in protective SzP proteins of Streptococcus zooepidemicus. AB - OBJECTIVES: To characterize, on a molecular basis, variable regions of the SzP proteins of the Moore and Bryans serovars of Streptococcus zooepidemicus and specificity of opsonic responses. SAMPLE POPULATION: 14 Moore and Bryans serovars of S zooepidemicus. PROCEDURE: Using polymerase chain reaction analysis and primers from the 5' and 3' sequences of the prototype gene SzPW60, the SzP genes of each Moore and Bryans serovar were sequenced and translated, then the amino acid sequences were compared. RESULTS: Comparison of the amino acid sequences revealed 2 variations at the N terminus; a hypervariable (HV) region from residue 106 to 166, approximately; and proline-glutamic acid-proline-lysine repeats in the carboxy terminus that ranged in number from 7 to 12. Five distinct motifs, HV 1 to 5, which varied independently of the N termini were found in the internal HV region. All serovars were opsonized by antiserum to the prototype SzPW60 protein, indicating that opsonogenic epitopes are on the conserved regions of the protein. CONCLUSION AND CLINICAL RELEVANCE: Variant motifs may be valuable in epizootiologic and pathogenesis studies of S zooepidemicus infections of the respiratory tract of young horses and in determining whether there are populations of S zooepidemicus unique to specific animal hosts. It is also clear from the opsonic responses to SzP that at least a portion of the protective responses are probably not serovar specific. PMID- 9736392 TI - Cardiorespiratory and anesthetic effects of propofol and thiopental in dogs. AB - OBJECTIVE: To compare cardiorespiratory and anesthesia effects of IV administered propofol and thiopental in dogs. ANIMALS: 6 healthy mixed-breed dogs. PROCEDURE: Each dog was anesthetized with isoflurane, then a thermistor catheter was inserted in the pulmonary artery. After a minimum of 2.5 hours of recovery, a catheter was placed in a cephalic vein for administration of lactated Ringer's solution and drugs. Propofol (8 mg/kg of body weight) or thiopental (19.4 mg/kg) was administered to each dog in a randomized crossover design study. All dogs were intubated and allowed to breathe 100% oxygen spontaneously. Heart rate and rhythm; systolic, diastolic, and mean arterial blood pressures; respiratory rate; end-tidal carbon dioxide concentration; tidal volume; and reflexes (toe web pinch, palpebral response, and jaw tone) were measured before and every 2 minutes for the first 10 minutes, then at 15, 30, and 60 minutes after drug administration. Cardiac output was determined at 0, 2, 6, 10, 15, 30, and 60 minutes, and blood samples were collected at 0, 2, 10, and 30 minutes. Time to endotracheal extubation, head lift, and ability to sit sternally and walk unaided were recorded. RESULTS: 3 of 6 dogs in each group were apneic after drug administration. Reflexes were decreased similarly for both anesthetic agents, but were not completely lost. Time to sternal position and walking unaided were significantly shorter in response to propofol. CONCLUSION: Anesthesia was rapid; however, respiratory depression and apnea were major adverse effects associated with propofol and thiopental. Propofol has the advantage of inducing rapid, coordinated anesthesia recovery. PMID- 9736391 TI - Efficacy of a drug combination of praziquantel, pyrantel pamoate, and febantel against giardiasis in dogs. AB - OBJECTIVE: To evaluate efficacy of a combination of praziquantel, pyrantel pamoate, and febantel at 2 dosages for treating naturally acquired giardiasis in dogs. ANIMALS: 6 male and 9 female Beagles. PROCEDURE: Dogs were identified as naturally infected with Giardia sp, using the zinc sulfate concentration technique (ZSCT), and were allocated to 1 of 3 groups. Group-1 dogs were treated orally with a praziquantel (5.4 to 7 mg/kg of body weight), pyrantel pamoate (26.8 to 35.2 mg/kg), and febantel (26.8 to 35.2 mg/kg) combination, every 24 hours for 3 doses. Group-2 dogs were treated with the combination once. Group-3 dogs were nontreated controls. Four fecal samples were examined, using the ZSCT, from each dog of each group within 6 days of the last treatment. Dogs were considered to have giardiasis if 1 or more of the fecal samples had positive results for Giardia cysts. Dogs were examined daily for at least 10 days after the last treatment. RESULTS: Giardia cysts were not detected in the feces of any group-1 dog or in the feces of 2 of 5 group-2 dogs. Cysts were detected in the feces of 5 of 5 group-3 (nontreated control) dogs. Signs of toxicosis were not observed in any dog. CONCLUSION AND CLINICAL RELEVANCE: The current labeled dose (for treatment of various nematodes and cestodes, but not Giardia sp) of the combination given orally once reduces cyst excretion in Giardia-infected dogs, and should be considered for treatment of dogs shedding Giardia cysts, whether or not they have clinical signs of infection. PMID- 9736393 TI - Expression and bioactivity of recombinant canine erythropoietin. AB - OBJECTIVE: To produce recombinant canine erythropoietin (rcEPO) and compare its biological activity with that of recombinant human EPO (rhEPO). ANIMALS: C57BL/6J mice. PROCEDURE: The gene encoding cEPO was isolated from a genomic library and subcloned into an eucaryotic expression vector. Production of rcEPO was achieved by stable transfection of the expression construct into Chinese hamster ovary cells. Biological activity was evaluated in vitro by analyzing the mitogenic activity of rcEPO on murine erythroid progenitor cells. In vivo bioactivity was assessed in mice by measuring the ability of rcEPO to increase blood reticulocyte counts. RESULTS: Size and glycosylation of rcEPO expressed in Chinese hamster ovary cells were similar to values for commercial rhEPO. Canine and human EPO stimulated proliferation of murine erythroid progenitor cells in vitro and murine reticulocytosis in vivo in a dose-dependent manner. CONCLUSIONS: Comparable biological activity was observed for rcEPO and rhEPO in the 2 murine-based assay systems studied. By avoiding interspecies variation in protein structure and the resulting potential for immunogenicity, rcEPO should represent a better option than rhEPO for treatment of dogs with erythropoietin-dependent anemia. CLINICAL RELEVANCE: Therapeutic use of rhEPO in companion animals is limited by its immunogenicity and the resulting potential to induce pure red cell aplasia. Development and availability of species-specific EPO preparations should avoid this problem. PMID- 9736394 TI - Anti-inflammatory effects of topically applied dimethyl sulfoxide gel on endotoxin-induced synovitis in horses. AB - OBJECTIVE: To evaluate the effect of topically applied dimethylsulfoxide (DMSO) on lipopolysaccharide (LPS)-induced synovitis in the mid-carpal joint. ANIMALS: 6 sound, healthy, adult horses (12 carpi). PROCEDURE: In a double-blinded, crossover, paired study with a 1-week washout period, mid-carpal joints were allocated to group 1 (DMSO, n = 6) or group 2 (control, n = 6). Each joint was injected with 1.3 ml (0.0125 ng/dl) of LPS to induce synovitis. For group-1 joints, DMSO gel (15 g; 90%) was applied after injection of LPS and at 12-hour intervals for 60 hours. Joints of group 2 received LPS, but not DMSO gel. All horses were evaluated by serial lameness examinations and synovial fluid analyses (total and differential WBC count and total protein concentration) at 12-hour intervals for 60 hours after LPS injection. Plasma and synovial fluid were obtained at baseline and 36 hours to document presence of DMSO. RESULTS: Mean WBC concentration was significantly (P < 0.05) lower in group-1, compared with group 2 joints, at 24 hours and had a trend to be lower at 36 hours. Mean total neutrophil count was significantly lower in group-1, compared with group-2 joints at 24 hours. In group-1 joints, DMSO was detected by use of gas chromatography in the synovial fluid of 5 of 6 joints and in plasma from 1 of 6 horses. CONCLUSION: Topically applied DMSO penetrated into synovial fluid in sufficient quantities to be detected and to decrease joint inflammation. PMID- 9736395 TI - Histamine-induced adherence and migration of equine eosinophils. AB - OBJECTIVES: To examine effects of histamine on equine eosinophil adherence in vitro and to determine the histamine receptor subtype(s) and cell surface adhesion molecules that mediate this response. In addition, to determine the receptor subtypes involved in histamine-induced eosinophil migration. ANIMALS: 8 healthy ponies. PROCEDURE: Effects of histamine on equine eosinophil adherence to serum- or fibronectin-coated plastic, and migration in a microchemotaxis assay were examined. In some experiments, eosinophils were pretreated with histamine receptor antagonists or monoclonal antibodies raised against cell adhesion molecules. For comparison, the effect of histamine on equine neutrophil adherence and migration was studied. RESULTS: Histamine induced adherence of equine eosinophils, but not neutrophils, to serum- and fibronectin-coated plastic (P < 0.01). Histamine also caused migration of equine eosinophils, but not neutrophils (P < 0.01). Histamine-induced adherence and migration of equine eosinophils were inhibited by histamine, (H,)-receptor antagonists chlorpheniramine and mepyramine (P < 0.01), but not H2- or H3-receptor antagonists cimetidine and thioperamide. Monoclonal antibodies raised against CD18, but not very late antigen 4, reduced histamine-induced equine eosinophil adherence to serum- and fibronectin-coated plastic (P < 0.01). CONCLUSIONS: When released from mast cells or basophils, histamine could stimulate adherence and migration of equine eosinophils via H, receptor activation and induce adherence of equine eosinophils to opsonized surfaces or dermal connective tissue matrix proteins via CD18 activation. CLINICAL RELEVANCE: Histamine may have a part in regulating equine eosinophil function during parasitic killing or antigen-induced responses in horses with insect hypersensitivity. PMID- 9736396 TI - Physiologic response to dobutamine infusion during cardiac stress testing of dogs. AB - OBJECTIVE: To evaluate response of various cardiovascular variables after administration of incremental doses of dobutamine in healthy conscious dogs, using standardized dobutamine stress echocardiography (DSE). ANIMALS: 8 healthy dogs. PROCEDURE: A DSE was performed twice on each dog within 24 hours. Dobutamine was infused at a rate of 12.5 to 42.5 microg/kg/min, using incremental increases of 10 microg/kg/min. Doppler sphygmomanometry, electrocardiography, and echocardiography were performed. Left ventricular size, global ventricular performance, and left ventricular systolic myocardial function were measured by means of echocardiography. RESULTS: At the highest dosage, dobutamine induced an increase of 20+/-3% and 109+/-12% in systolic blood pressure and cardiac index, respectively. The latter was associated with a significant increase in heart rate and stroke index. Fractional shortening of the left ventricle, fractional thickening of the left ventricular free wall and interventricular septum, ejection fraction, and mean velocity of fiber shortening had a progressive and significant increase during dobutamine infusion. Preejection period and left ventricular ejection time had a progressive and significative decrease during the stress test. CONCLUSIONS: The technique used was feasable, safe, and repeatable in healthy conscious dogs. Control values were determined. CLINICAL RELEVANCE: Data for these healthy dogs might be useful for comparison with results obtained from dogs with known or suspected cardiovascular disease. PMID- 9736397 TI - Role of nitric oxide in in vitro contractile activity of the third compartment of the stomach in llamas. AB - OBJECTIVE: To determine the role of nitric oxide and an apamin-sensitive nonadrenergic-noncholinergic inhibitory transmitter in in vitro contractile activity of the third compartment in llamas. SAMPLE POPULATION: Isolated strips of third compartment of the stomach from 5 llamas. PROCEDURE: Strips were mounted in tissue baths containing oxygenated Kreb's buffer solution and connected to a polygraph chart recorder to measure contractile activity. Atropine, guanethidine, and indomethacin were added to tissue baths to inhibit muscarinic receptors, adrenoreceptors, and prostaglandin synthesis. Responses to electrical field stimulation following addition of the nitric oxide antagonist Nwo-nitro-L arginine methyl ester (L-NAME) and apamin were evaluated. RESULTS: Electrical field stimulation (EFS) resulted in a reduction in the amplitude and frequency of contractile activity, followed by rebound contraction when EFS was stopped. Addition of L-NAME resulted in a significant reduction in inhibition of contractile activity. Addition of apamin also resulted in a significant reduction in inhibitory contractile activity at most stimulation frequencies. The combination of L-NAME and apamin resulted in a significant reduction in inhibition at all frequencies. CONCLUSION: Nitric oxide and a transmitter acting via an apamin-sensitive mechanism appear to be involved in inhibition of contractile activity of the third compartment in llamas. CLINICAL RELEVANCE: Results suggest that nitric oxide plays an important role in mediating contractile activity of the third compartment in llamas. Use of nitric oxide synthase inhibitors may have a role in the therapeutic management of llamas with lesions of the third compartment. PMID- 9736398 TI - Synergistic effect of hydrochloric acid and bile acids on the pars esophageal mucosa of the porcine stomach. AB - OBJECTIVES: To determine effects of finely ground diet and food deprivation on pH and bile acid concentration in the proximal portion of the porcine stomach and effects of bile acids and pH on the pars esophageal mucosa in vitro. ANIMALS: Sixteen 15- to 30-kg pigs. PROCEDURES: Gastric content samples obtained from pigs fed a finely ground pelleted or coarsely ground meal diet were assayed for gastric pH and bile acids. Stratified squamous epithelium was studied in an Ussing chamber, and histologically. Electrical conductance and transmucosal mannitol fluxes (as indices of tissue permeability) were determined at pH 4.0, 2.0, and 1.5 and in response to treatment with 0, 1, 2, or 3 mM taurodeoxycholate or glycocholate. RESULTS: Pigs fed the finely ground feed had significantly (P = 0.01) lower proximal stomach pH than did pigs fed the coarse meal. Proximal stomach bile acids concentration was significantly (P = 0.04) higher in pigs fed the finely ground diet. The H+ and bile acids concentration increased with time after feeding. In vitro exposure of the stratified mucosa to high H+ (pH < 4.0) and bile salt concentration (> or = 1.0 mM) resulted in significant (P < 0.05) dose-dependent increase in tissue conductance and mannitol fluxes, whereas low pH or bile acids alone had little effect. CONCLUSIONS: High H+ and bile acids concentration in the stomach of pigs fed finely ground diets or subjected to feed deprivation may contribute to ulceration of the pars esophageal tissue. Bile acids act synergistically and in dose-dependent manner, with low pH causing damage to the stratified squamous epithelium in vitro. PMID- 9736399 TI - Evaluation of treatment with a pulsed electromagnetic field on wound healing, clinicopathologic variables, and central nervous system activity of dogs. AB - OBJECTIVE: To evaluate effects of treatment with a pulsed electromagnetic field (PEMF) on healing of open and sutured wounds, clinicopathologic variables, and CNS activity of dogs. ANIMALS: 12 adult female Beagles. PROCEDURE: Open and sutured wounds were created in the skin of the trunk of the dogs. Dogs were divided into 2 groups. One group received PEMF treatment and 1 group served as untreated (control) dogs. The PEMF-treated dogs received treatment twice a day starting the day before surgery and lasting through day 21 after surgery. Wounds were evaluated by use of tensiometry, planimetry, laser Doppler perfusion imaging, and histologic examination. Clinicopathologic variables and electroencephalographic tracings were also evaluated. RESULTS: Use of PEMF treatment resulted in significantly enhanced epithelialization of open wounds 10 and 15 days after surgery. Five days after surgery, wounds of control dogs had a negative value for wound contraction, whereas PEMF-treated wounds had a positive value. The PEMF treatment did not cause significant changes in short-term planimetric, perfusion, tensiometric, histologic, clinicopathologic, or electroencephalographic results. CONCLUSIONS: The PEMF treatment enhanced wound epithelialization in open cutaneous wounds and provided indications of early contraction without significant short-term changes in other variables. PMID- 9736400 TI - Isolation and chondrocytic differentiation of equine bone marrow-derived mesenchymal stem cells. AB - OBJECTIVE: To isolate mesenchymal stem cells from adult horses and determine specific monolayer culture conditions required to enhance biochemically and phenotypically defined chondrocytic differentiation. ANIMALS: 2 adult horse bone marrow donors without skeletal or hematologic abnormalities. PROCEDURE: Bone marrow was aspirated from the sternebra, and mesenchymal stem cells were isolated by centrifugation and cultured in monolayers. Subcultures were established in 24 well plates on day 13. Culture medium was harvested every 2 days, and culture of 12 of the 24 wells was terminated on day 6 and of the remaining wells on day 12. Medium proteoglycan content was determined for all samples, and proteoglycan monomeric size was determined for pooled samples from days 2-6 and 8-12. Total nucleated cell numbers were determined at culture termination on days 6 and 12. Histologic, histochemical, and collagen immunohistochemical analyses of multiwell chamber slides harvested on day 6 or 12 were performed. RESULTS: Mesenchymal cells were an abundant cellular constituent of bone marrow aspirates, and separation of hematopoietic elements was achieved by centrifugation and delayed medium exchange. The remaining mesenchymal stem cells progressed from large, spindyloid, fibroblastic-appearing cells to a rounder shaped cell which formed colony plaques; isolated cells remained more spindyloid. Mesenchymal cell transformation toward a chondrocytic phenotype was verified by a shift in expression from collagen type I to type II, and an increase in quantity and molecular size of proteoglycans synthesized over time. CONCLUSIONS: Mesenchymal stem cells obtained from adult horses have the capacity to undergo chondrogenic differentiation in monolayer cultures and may provide a locally recruitable or transplantable autogenous cell source for articular cartilage repair. PMID- 9736401 TI - Effect of three types of half-limb casts on in vitro bone strain recorded from the third metacarpal bone and proximal phalanx in equine cadaver limbs. AB - OBJECTIVE: To determine effect of 3 half-limb casts on bone strains recorded from the proximal phalanx (P-1) and third metacarpal bone (MCIII) of equine cadaver limbs, using a mechanical testing machine. ANIMALS: 12 equine cadaver limbs and 4 live horses. PROCEDURE: Bone strains were recorded at middorsal P-1 and the dorsal cortical aspect of the distal third of MCIII while limbs were variably loaded with 100 to 1,000 lb of force. To determine ability of the cast to protect the distal portion of the limb from weight-bearing loads, strains were recorded with the limb in 1 of the 3 casts and with it unsupported. To determine cast induced discomfort, weight-supporting and transfixation pin casts were evaluated on 2 live horses RESULTS: All 3 casts significantly reduced bone strain at P-1. Significant differences were observed: mean 61% reduction for the standard half limb cast, 84% for the transfixation pin cast, and 97% for the weight-supporting cast at weight-bearing force of 500 lb. Only the weight-supporting cast significantly reduced strains recorded from MCIII. The weight-supporting cast was not well tolerated by 2 live horses. CONCLUSIONS: The 3 casts significantly reduced transfer of weight-bearing forces to the distal portion of the limb. The weight-supporting cast effectively reduced strain on the P-1 to near 0, but was well tolerated by live horses. The transfixation pin cast reduced strain on the P 1 by > 80% at weight-bearing loads of 500 lb, and live horses were comfortable. Standard half-limb casts significantly reduced strains on the P-1, but to a lesser degree than did other casts. PMID- 9736402 TI - Genetic alterations in primary and secondary hyperparathyroidism. AB - Hyperparathyroidism refers to a term representing a wide spectrum of parathyroid disorders that are characterized by the increased production of parathyroid hormone. Hyperparathyroidism was once thought to be rare but is now more commonly recognized, affecting 1 in 500 women over 40 years of age. Yet the interpretation of parathyroid pathology is still controversial and confusing. Over the past 10 years, genetic changes (ret and menin genes) involved in the pathogenesis of MEN 2 and MEN 1 have been discovered in succession. Different mutations of the calcium-sensing receptor gene have been identified in neonatal severe hyperparathyroidism and familial hypocalciuric hypercalcemia, respectively. The HRPT 2 gene responsible for the development of hereditary hyperparathyroidism and jaw tumors has been localized on the 1q21-31 locus. Several genetic alterations have also been characterized in primary and secondary hyperparathyroidism. Different genetic alterations appear to involve the development of different types of hyperparathyroidism. These novel advances give us new insights into the pathogenesis of hyperparathyroidism and allow better differentiation between the different types of parathyroid disorders. PMID- 9736403 TI - Localization of identified advanced glycation end-product structures, N epsilon(carboxymethyl)lysine and pentosidine, in age-related inclusions in human brains. AB - The recent identification of age-related accumulation of advanced glycation end products (AGE) of the Maillard reaction in neurons and vessels of the human brain suggests the involvement of AGE in the aging process. A variety of inclusions such as lipofuscin granules, corpora amylacea, Hirano bodies, granulovacuolar degenerations and ubiquitin-positive granular structures are found in the aged human brain. These age-related inclusions contain insoluble and non-degradable proteins. Advanced glycation end-product-modified proteins are also known to be insoluble and protease resistant. The similarity between proteins in such inclusions and AGE-modified proteins suggests the presence of AGE in inclusions. To investigate this possibility, the presence of two known AGE structures, N epsilon(carboxymethyl)lysine (CML) and pentosidine, was examined in age-related inclusions. Immunohistochemical examination of the medial temporal area of brain tissues obtained at autopsy from seven non-demented elderly individuals demonstrated positive reactions in lipofuscin granules and corpora amylacea but not in other inclusions for anti-CML and anti-pentosidine antibodies. As CML and pentosidine are glycoxidation products among AGE, the results suggest that glycation and/or oxidation may be involved in the formation of lipofuscin granules and corpora amylacea. PMID- 9736404 TI - Immunohistochemical overexpression of p16 protein associated with intact retinoblastoma protein expression in cervical cancer and cervical intraepithelial neoplasia. AB - Both p16 and retinoblastoma (Rb) proteins are important tumor suppressors that regulate the cell cycle. The status of both proteins in invasive cervical cancer and cervical intraepithelial neoplasia (CIN) has not yet been examined. The aim of this study was to investigate the expression of p16 and Rb proteins by immunohistochemistry using 98 formalin-fixed and paraffin-embedded samples of various cervical neoplastic lesions. Strong immunoreactivity for the p16 protein was observed in both the nuclei and cytoplasm of all CIN and invasive cancer cases except several low-grade CIN lesions. Expression of Rb protein was also demonstrated in the scattered nuclei of neoplastic and normal cells in all cases investigated. The results suggest that the deletion or mutational inactivity of both p16 and Rb proteins may be a rare event in cervical carcinogenesis. Moreover, overexpression of the p16 protein may be a useful diagnostic marker for cervical neoplastic lesions on routine laboratory screening. PMID- 9736405 TI - Microsatellite instability in young patients with colorectal cancer. AB - Genetic instability is closely correlated to the pathogenesis of hereditary non polyposis colon cancer (HNPCC), which is clinically characterized by a family history and early onset. To investigate the role of genetic instability in young patients with colorectal cancer (CRC), 22 CRC patients, who were aged younger than 30 at the time of diagnosis, were studied. Patients with familial adenomatous polyposis were excluded. Among the 22 cases, seven were identified as microsatellite instability positive (MI+), and more than five microsatellite markers among the 15 tested markers showed an additional band pattern in the tumor tissue. None of the remaining 15 cases showed instability in any microsatellite marker. Two of seven MI+ cases were classic HNPCC. While all MI+ cases had one or no metastatic lymph node, 53.3% of MI- cases showed metastasis in two or more regional lymph nodes. Allelic deletion of the 17p12-13 chromosome around the p53 locus occurred in 16.7% of MI+ cases, and 80.0% of MI- cases showed loss of heterozygosity at that locus. hMSH2 Protein expression, assessed by immunohistochemistry, was absent in two cases, both of which were MI+. When we tested two to four sites of MI+ tumors, transforming growth factor beta receptor type II was mutated in a homogeneous pattern in five MI+ cases. In addition, frame-shift mutations of BAX, insulin-like growth factor II receptor, hMSH3 and hMSH6 were found in three cases, five cases, five cases and one case, respectively. In contrast to the consistent mutation of the transforming growth factor-beta receptor type II gene, mutations of other genes varied in different portions of the tumors. PMID- 9736406 TI - Gliosarcoma: an immunohistochemical, ultrastructural and fluorescence in situ hybridization study. AB - Three cases of primary gliosarcoma (GS) were studied by immunohistochemical, ultrastructural and fluorescence in situ hybridization (FISH) methods. All tumors occurred in the supratentorial regions of the body. No patient had a prior history of irradiation to the brain. All patients died of tumor within 1 year, and autopsies were performed in two cases. Microscopically, each of the three tumors showed a mixture of glioblastoma (GBM) and a sarcomatous component (SC), which resembled fibrosarcoma with various histological features. Numerous collagen and reticulin fibers were seen in the SC of all tumors. Glial fibrillary acidic protein (GFAP) was immunoreactive only in the gliomatous component (GC). Factor VIII-related antigen was negative except for endothelial cells. One tumor exhibited alpha-smooth muscle actin positivity in the SC. Expression of MIB-1 and p53 protein was demonstrated in both components for all tumors. Labeling indices (LI) for MIB-1 ranged from 7.7 to 36.1%, and LI for p53 protein ranged from 2.9 to 57.0%. Ultrastructurally, astrocytic cells were characterized by a polygonal configuration with many cytoplasmic projections and occasional filaments. Spindle shaped fibroblasts in the SC contained well-developed rough endoplasmic reticulum. Fluorescence in situ hybridization (FISH) performed on fresh materials or paraffin-embedded tissue demonstrated single signals for chromosome 10 in 40.6 58.3% of cells and for chromosome 17 in 37.9-48.6% of cells. Two tumors were regarded as containing losses of both chromosomes 10 and 17, while the third showed a substantial loss only of chromosome 10. As similar aberrations have been reported in GBM, these chromosomal abnormalities suggest a common pathogenesis in GS and GBM. PMID- 9736407 TI - Collagen and elastin synthesis by desmoid tumor in vitro. AB - In order to characterize human desmoid tumors in vitro, the production of collagen and elastin and the expression of collagen types alpha1(I), alpha1(III) and transforming growth factor (TGF)-beta1 mRNA were investigated in six desmoid tumors; five derived from familial adenomatous polyposis patients and one from a sporadic case. The proportion of collagen production to total protein production was determined by 3H-imino acid incorporation, an indicator of collagen synthesis, using high-performance liquid chromatography (HPLC). The proportion of collagen production to total protein production was much higher in all six desmoid tumors compared with human skin fibroblasts (HSF). Quantitatively, the rate of elastin synthesis in desmoid tumor cells monitored by valine-proline peptide was also significantly higher than in HSF. Pro-alpha1(I) collagen mRNA was highly expressed in both desmoid tumors and HSF at approximately the same level, whereas pro-alpha1(III) collagen mRNA was more abundant in some of the desmoid tumors than the normal skin fibroblastic cell lines. Tumor growth factor beta1 mRNA, which is believed to stimulate collagen synthesis, was expressed in both desmoid tumors and HSF to the same extent. These results demonstrate the increased formation of collagen and elastin in desmoid tumors in vitro and suggest that the increased synthesis of elastin rather than of collagen and TGF beta1 may be involved in increased fibrogenesis by desmoid tumors. PMID- 9736408 TI - Schwannoma of the liver: report of two surgical cases. AB - Two surgical cases of benign schwannoma of the liver were examined: A 64-year-old female who was admitted to hospital for the treatment of early gastric cancer (case 1) and a 69-year-old female who had pitting edema in the legs (case 2). Diagnostic imaging revealed large solid masses located in the caudate lobe through to the left lobe of the liver in case 1 and in the left lobe in case 2. Both patients showed no signs of von Recklinghausen's disease. Partial hepatectomies were performed and the maximum diameter of their resected hepatic tumors was 4 cm and 15 cm, respectively. Histologically, the well-demarcated tumors were yellowish in color, elastic-hard in consistency and consisted predominantly of short spindle-shaped cells proliferating in an interlacing fashion. The tumor lesions were separated by fibrous bands and surrounded by a lymphoid cuff. Immunohistochemically, the tumor cells were positive for S-100 protein. The tumors were diagnosed as benign hepatic schwannoma. PMID- 9736409 TI - Clear cell odontogenic tumor in the mandible: report of a case with an immunohistochemical study of epithelial cell markers. AB - A rare case of clear cell odontogenic tumor is presented with an immunohistochemical study using epithelial cell markers. A 35-year-old Japanese man was admitted with a complaint of painless swelling in the anterior region of his mandible. Radiological examination showed a relatively well-defined multilocular radiolucency with root resorption of the adjacent teeth. Despite a subtotal mandibulectomy, the tumor recurred three times. Histologically, the tumor was composed of proliferating clear cells and infiltrated through the cancellous bone. Histochemical and ultrastructural analyses detected cytoplasmic glycogen granules in the clear cells. They showed immunoreactivities for cytokeratin 8, 13 and 19, filaggrin and anti-ameloblastoma antibodies, suggesting an odontogenic epithelial origin. PMID- 9736410 TI - Cystic renal cell carcinoma: arose as a simple cyst, recurred repeatedly as a multicystic mass and finally presented as a solid mass. AB - An unusual case of cystic renal cell carcinoma in a 70-year-old Japanese male is reported. He had had a simple renal cyst removed 1 year ago. On presentation the right kidney was surrounded by multiple translucent cysts, which varied in size from 1 to 50 mm. The cyst walls were lined by single-layered cuboidal epithelium. The differential diagnosis included cystic mesothelioma, cystic lymphangioma and multicystic dysplastic kidney. An immunohistochemical and ultrastructural study and flow cytometric analysis of DNA ploidy were performed. The tumor was differentiated to such an extent that it was difficult to diagnose as carcinoma; however, it recurred repeatedly. Three and a half years after initial presentation the tumor had invaded the ileum with the pathological change to be almost solid when viewed grossly and, microscopically, showed tubulo-papillary structures in addition to a cystic pattern. The DNA ploidy pattern revealed a near-diploid aneuploid in the early specimen and a polyploid aneuploid in the last specimen. PMID- 9736411 TI - Oncocytic cystadenoma of the parotid gland with prominent signet-ring cell features. AB - A case of distinctive benign cystadenoma of the parotid gland composed of several different morphological components is presented. The most conspicuous morphological component and the largest part of the neoplasm was represented by solid sheets of oncocytic cells surrounded by myoepithelial cell layer. Most oncocytic cells possessed large intracytoplasmic vacuoles with the nuclei displaced towards the periphery, imparting them with a striking signet-ring cell appearance. The size of the intracytoplasmic vacuoles ranged from 4 to 50 microm. Immunohistochemically these signet-ring cells lacked immunoreactivity for S-100 protein and cytokeratin but they strongly stained for antimitochondrial antibody 113-1. The present case illustrates an unusual, hitherto undescribed, morphological feature of benign oncocytic cystadenoma of the parotid gland. PMID- 9736412 TI - A case of double primary adenocarcinoma of the lung with multiple atypical adenomatous hyperplasia. AB - A case of double primary adenocarcinoma of the lung with multiple atypical adenomatous hyperplasia (AAH) in a 77-year-old woman is reported. Histopathologically, in the resected left upper lobe of the lung, both cancers were diagnosed as well-differentiated papillary adenocarcinoma, and 161 lesions of AAH were also found. Both the cancer lesions and six AAH (greater than 3 mm in diameter) were examined with regard to immunoreactivity of carcinoembryonic antigen (CEA) and p53 gene product, microsatellite instability (MI) and loss of heterozygosity (LOH) on chromosome 9q and 17q by polymerase chain reaction (PCR). Although both cancers expressed CEA, they did not show clonal immunoreactivity for the p53 gene product. Atypical adenomatous hyperplasia expressed CEA weakly and showed no immunoreactivity for p53 gene protein. Both carcinomas showed LOH on chromosome 17q, and one of them showed LOH on chromosome 9q. In six AAH, LOH on chromosome 17q was detected in two tumors, and one of them also showed LOH on chromosome 9q. One AAH, which was negative for LOH on chromosome 17q and 9q, showed MI at D17S791. These results indicated that AAH is a clonal neoplastic lesion with genetic abnormalities and should be called intraepithelial pneumocyte neoplasia, and that each of the numerous papillary lesions in this case was considered to be an independent lesion. PMID- 9736413 TI - Two cases of mesothelial/monocytic incidental cardiac excrescences of the heart. AB - Two cases of mesothelial/monocytic incidental cardiac excrescences in a 66-year old female and an 80-year-old male are presented. Lesions had solid and tubular pattern formations which were composed of two predominant cell types of histiocytoid cells and cuboidal cells arranged in strips. The histiocytoid cells were round and had well-defined nuclei with prominent nuclear grooves. They had a low nuclear to cytoplasmic ratio. There were no atypical mitoses. Immunohistochemically, these cells were positive for leukocyte common antigen (LCA) and CD68 (KP-1) but negative for keratin. The cuboidal cells were present in strips, had haphazardly arranged surface microvilli and had small round non cleaved nuclei. These cells were positive for keratin but negative for LCA, CD68, p53, proliferative cell nuclear antigen, alpha-smooth muscle actin, Factor VIII, epithelial membranous antigen and vimentin. These lesions are probably reactive because of their heterogeneous components; an expected feature for an essentially artifactual lesion that is related to cardiac surgery and invasive catheterization. Immunohistochemical studies are useful for avoiding misdiagnosis of neoplasms. PMID- 9736414 TI - Ovarian mucinous cystadenocarcinoma with malignant mural nodules. AB - A case of ovarian mucinous cystadenocarcinoma with malignant mural nodules is reported. The patient was a 28-year-old Japanese female (gravida 0, para 0) with a 2 year history of increasing abdominal fullness and edema of the lower extremity. Physical examination showed a large mass in the abdomen. An abdominal ultrasound and computed tomography demonstrated a multilocular cyst with a solid component. Lymph node and distant metastases were not found. These tests were followed by surgery. The resected right ovarian tumor measured 25 x 22 x 18 cm. On cut sectioning, it was multilocular with multiple mural nodules. Microscopically, the cyst wall was lined with papillary infoldings of atypical mucinous epithelium (intestinal type). Nuclear stratification, cribriform and back-to-back glandular patterns and stromal invasion were observed. In addition, the solid area of the mural nodules showed spindle or polygonal cells with increased mitotic activity including atypical mitoses. Nuclear pleomorphism was marked. Necrosis and hemorrhage were also present. Reticulin stain showed these pleomorphic cell clusters circumscribed by reticulin fibers and these cells were immunoreactive for vimentin and p53. PMID- 9736415 TI - Adenosarcoma of the uterine cervix presenting as a cervical polyp. AB - A case of adenosarcoma arising from the uterine cervix of a 55-year-old female who complained of vaginal discharge is reported. A polyp, 6 x 2 x 1.5 cm in size, identified in the cervical canal was clinically diagnosed as benign cervical polyp and resected. Histologically, the polyp was composed of benign epithelial components and sarcomatous stroma wherein periglandular hypercellularity and some mitoses including atypical ones were noted. Immunohistochemically, stromal cells were positive for muscle-type actins, desmin and estrogen receptor. Adenosarcoma is a rare, biphasic tumor of the uterus and usually presents as a polypoid mass in the endometrial cavity. When they arise from the cervix, adenosarcomas may be confused with benign cervical polyps clinically and pathologically. As the former often recurs, microscopic differentiation is crucial for further treatment. PMID- 9736416 TI - Controlled-release products for the control of the estrus cycle in cattle, sheep, goats, deer, pigs, and horses. AB - This paper describes the estrus cycles of a number of livestock breeds and reviews the controlled-release drug delivery systems that are currently available for the purpose of controlled breeding. The bovine estrus cycle is reviewed in detail, and the estrus cycles of other species are described in a manner that highlights similarities and differences between species. Pertinent formulation and pharmacokinetic information about current drug delivery systems is presented and discussed, and recent advances in this area are also described. PMID- 9736417 TI - Bioerodible polymers for ocular drug delivery. AB - Development of ophthalmic drug-delivery systems has always been challenging. The commonly used route for drug delivery to the anterior segment of the eye has been the conjunctival cul-de-sac. Because of drawbacks associated with this route, new approaches have been investigated for delivery of drugs to the eye by means of polymeric delivery systems. Development of controlled drug-release devices has been a major step forward in this respect. Bioerodible polymers have been at the forefront of such systems. They are very important because they eliminate the need for removing the implant after complete drug release. Bioerodible polymers have been divided into three classes based on their mechanism of hydrolysis: Type I--hydrolysis of crosslinked hydrogels; Type II--solubilization by ionization or hydrolysis of linear polymers; and Type III--biodegradation by backbone cleavage. Polymers from all three classes are discussed in detail in this review. PMID- 9736418 TI - Inhibition of chronic vessel wall intimal hyperplasia following acute anticoagulant treatment: relative effects of heparin and dermatan sulphate. AB - Surface-bound thrombin which contributes to vessel wall hyperplasia, is resistant to inhibition by heparin/antithrombin III (/ATIII) but not to inhibition by dermatan sulphate/heparin cofactor II (/HCII). To determine the effects of heparin and dermatan sulphate on vessel wall hyperplasia after a first or second injury, rabbit carotid arteries first were injured, using a standard procedure (first injury). Half of the first-injury rabbits were given heparin, dermatan sulphate, or saline, 5 minutes before and at 30-minute intervals over 2 hours post-injury, and then allowed to recover. Four weeks later, the first-injury treated animals were killed and their injured carotid arteries were processed histologically. The remaining untreated first-injury rabbits were also allowed to recover. At 4 weeks, those rabbits were re-anesthetized and their first-injury arteries (which were occluded >75%), were isolated, and vessel wall lumen patency was re-established by endarterectomy (second injury). During this second injury, the animals were treated with heparin, dermatan sulphate, or saline as described above. Four weeks after this second injury, these rabbits were killed and their second injury arteries were processed histologically. Intimal hyperplasia determined histologically, was expressed as an x-fold increase in vessel wall cross-sectional area (i.e., [(media+intima area) media area]). Vessel wall lumen occlusion was expressed as [1-(lumen area/internal elastic lamina area) x 100; % occlusion]. Vessel wall area in the saline-treated animals, increased 2.6+/-1.2 and 2.4+/-1.0 fold respectively, means+/-SD, n = 12, within 4 weeks of the first and second injuries. These increases were due to intimal hyperplasia and associated with 75+/-19% and 79+/-21% occlusion of the vessel wall lumen, respectively. Heparin had little effect, whereas dermatan sulphate (1) decreased hyperplasia by 45% after the first injury and by 47% after the second injury, p<0.008 and <0.03, respectively, and (2) decreased vessel wall occlusion 47+/-12% and 33+/-5% after the first and second injury, respectively. We conclude that (1) dermatan sulphate/HCII may be a useful inhibitor of vessel wall hyperplasia following vessel wall injury, and (2) this effect can be achieved by an acute anticoagulant treatment at the time of injury, unlike heparin/ATIII. PMID- 9736419 TI - Plasma lipoprotein(a) levels are high in patients with central retinal artery occlusion. AB - High plasma lipoprotein(a) (Lp[a]) concentration is an independent risk factor for atherosclerosis and thrombosis. To study the implications of Lp(a) in central retinal artery occlusion (CRAO), we examined Lp(a) levels and molecular weights (MWs) of apolipoprotein(a) (apo(a)). Mean Lp(a) concentration was significantly higher in the cases with CRAO than in the controls. Lp(a) levels higher than 30 mg/dl were also more frequent in the CRAO cases than in the controls. Lp(a) concentrations correlated significantly with low-MW isoforms of apo(a). Impaired fibrinolysis and atherogenesis induced by Lp(a) may play a role in the pathophysiology of CRAO. Since high Lp(a) levels were reported in CRVO by other investigators, patients with central retinal vein occlusion (CRVO) were also examined for Lp(a). Although Lp(a) levels were higher in the CRVO cases than in the controls, the difference was not significant. Therefore, high Lp(a) levels may not be associated with venous thrombosis and/or embolism. PMID- 9736420 TI - Differential effects of low molecular weight heparin and unfractionated heparin on circulating levels of antithrombin and tissue factor pathway inhibitor (TFPI): a possible mechanism for difference in therapeutic efficacy. AB - There is growing evidence on the superior efficacy and safety of low molecular weight heparins (LMWHs) over unfractionated heparin (UFH) for the treatment of both venous and arterial thromboembolism. Heparin exerts its function by potentiating antithrombin and by mobilizing tissue factor pathway inhibitor (TFPI) into the circulation. The present study was conducted to compare the effect of subcutaneous LMWH and infusion of UFH on these two anticoagulants in plasma. Eighteen healthy male volunteers were randomly allocated to therapy with continuous intravenous (iv) UFH (n=6) (initial infusion rate 450 IU/kg/day) or low molecular weight heparin (LMWH) (enoxaparin, 1.5 mg/kg/day) subcutaneously (sc) once daily for 72 hours. Free TFPI antigen, measured by a solid-phase two site enzyme immunoassay, and antithrombin and protein C activities, measured by chromogenic assays, were assessed in plasma samples before, during, and after anticoagulant treatment. Infusion of UFH, but not subcutaneous LMWH, was found to attenuate the antithrombotic defense by a selective decrease of both circulating antithrombin (-21+/-7%, p<0.0001) and of free and endothelial-bound TFPI. The changes in antithrombin and TFPI by LMWH and UFH were statistically different between groups (p<0.001). The differential effect of UFH and LMWH on antithrombin and TFPI may explain the superior efficacy of subcutaneous LMWH compared with conventional intravenous UFH for treatment of both arterial and venous thrombosis. PMID- 9736421 TI - Neutrophil adhesion and activation during systemic thrombolysis in acute myocardial infarction. AB - In a pilot study, alterations of polymorphonuclear neutrophil function during systemic thrombolysis in acute myocardial infarction have been investigated in humans. The following parameters of neutrophil function were measured before and at 15 and 45 minutes after initiation of systemic thrombolysis with a recombinant tissue-type plasminogen activator in 20 patients with acute myocardial infarction: (1) neutrophil adhesion and (2) neutrophil activation. During systemic thrombolysis a significant decrease was observed in neutrophil adhesion (5.5+/-6.4 to 3.2+/-3.3; p<0.05), in phagocyting neutrophil activation (39+/-18 to 25+/-14%; p<0.05), and in resting neutrophil activation (9+/-7 to 3+/-4%; p<0.05). Successful reperfusion coincided with a significantly higher reduction of phagocyting neutrophil activation (40+/-14 to 20+/-12% vs. 39+/-24 to 26+/-19% in unsuccessful reperfusion; p<0.05), and of neutrophil adhesion (6.2+/-5.7 to 2.7+/-3.0 vs. 4.1+/-3.8 to 3.5+/-4.0 in unsuccessful reperfusion; p<0.05) during thrombolysis. Systemic thrombolysis in acute myocardial infarction is accompanied by a reduction in neutrophil adhesion and activation dependent on thrombolytic success. PMID- 9736423 TI - Dermatan sulfate enhances the lysis of laser-induced thrombus in vivo. PMID- 9736422 TI - Measurement of thrombomodulin mRNA expression in brain capillaries by polymerase chain reaction. AB - Thrombomodulin (TM), an endothelial integral membrane protein, is a potent activator of the protein C anticoagulant pathway. TM protein expression is limited and regionally distributed in the brain. Recent investigations have demonstrated low TM mRNA expression by brain endothelium, corresponding to its distribution at the protein level. To facilitate the study of TM expression at the transcriptional level, we measured TM mRNA by quantitative-competitive polymerase chain reaction (QC-PCR) and by standard densitometric analysis of reverse transcriptase-PCR products (RT-PCR) in different regions of bovine brain. QC-PCR demonstrated differential TM mRNA expression in the pons (100+/-9%), cerebellum (359+/-103%), and cortex (441+/-24%). We compared these results with those of RT-PCR and found similar differences in relative TM mRNA expression in the pons (100+/-44%), cerebellum (343+/-8%), and cortex (404+/-62%). Data derived by QC-PCR and RT-PCR were highly correlated (r=0.99, p<0.03). These findings indicate that either QC-PCR or RT-PCR can be used to accurately quantify TM mRNA. PMID- 9736424 TI - Treatment with interferon gamma enhances fibrinolysis in systemic sclerosis. PMID- 9736425 TI - Expression of p21 (Waf1/Cip1) in head and neck cancer in relation to proliferation, differentiation, p53 status and cyclin D1 expression. AB - p21(Waf1/Cipl) is a critical downstream effector in the p53-dependent pathway of growth control and causes growth arrest through inhibition of cyclin-dependent kinases. In this study 67% of 43 head and neck squamous cell carcinoma (HNSCC) and 60% of 15 tumour-adjacent oral dysplasias overexpressed p21 by immunohistochemical staining. Overexpression of p21 in HNSCC was independent of the presence of functional p53, as assessed by analysis of mutations and loss of heterozygosity and by immunohistochemisty. Rather, the expression pattern of p21 was associated with differentiation. Furthermore, in most tumours, the p21 positive cells did not incorporate bromodeoxyuridine (BrdU), which indicates inhibition of proliferation by p21 in these cells. In some tumours, p21 was also expressed in proliferating cells. In these latter tumour cells, cyclin D1 was frequently expressed as well. Therefore, we suggest that expression of cyclin D1 might overcome the inhibitory effect of p21 in these cells. PMID- 9736426 TI - Relationship of p53 overexpression to other cell cycle regulatory proteins in oral squamous cell carcinoma. AB - Aberrations of the p53 gene and the overexpression of its protein are described in a variety of neoplasms, including oral and other head and neck cancers. Here we report the association of p53 (over)expression with a downstream cell cycle inhibitor p21/waf 1 in oral squamous cell carcinoma (SCC). The loss of expression of p16 and p27, two other cyclin-dependent kinase (cdk) inhibitors, was also examined. In this panel of tumours, 10/24 carcinomas were p53-immunopositive. Heterogeneous expression of p21 and p27 was seen in 10/24 SCC and 9/16 SCC, respectively, and this was not correlated to p53 status. The expression of p21 and p27 in these SCCs suggests the existence of mechanisms by which some growing tumour cells may tolerate these cell cycle inhibitors; eight SCCs lacked expression of both inhibitors but only two of these cancers overexpressed p53, suggesting that accumulation of p21/p27 can be independent of the functional status of the p53 gene. Data do not support a clear example of a phenotype that shows an overexpression of p53 with downregulation of p21 or p27 leading to cell cycle alterations. Furthermore, only three SCCs were p16-negative and p53 positive. This suggests that these two tumour suppressors may act in separate pathways. PMID- 9736427 TI - Pre-operative radio-chemotherapy enhances apoptotic cell death in oral squamous cell carcinoma. AB - The effect of pre-operative radio-chemotherapy (RCT) has been examined in a total of 15 oral squamous cell carcinomas (SCCs), in terms of apoptosis (cell loss) and proliferation. All the patients received pre-operative radiation at a dosage of 30 or 40 Gy, as well as anticancer agents including tagaful (FT), 5-fluorouracil (5-FU), bleomycin (BLM) and peplomycin (PEP). Surgical specimens were obtained before and after RCT, and serial sections were prepared for immunohistochemistry for p53 oncoprotein and Ki-67 antigen, as well as for terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick end labeling (TUNEL). TUNEL indices (TI; percentage of TUNEL-positive cells in the tumor cells) before and after RCT were 1.2+/-1.1 and 4.7+/-2.9 in the nine well-differentiated oral SCCs, and 1.0+/ 0.7 and 3.9+/-2.1 in the six poorly differentiated SCCs, respectively. Similarly, Ki-67 indices (KI; percentage of Ki-67 antigen-positive cells in tumor cells) before and after RCT were 31.1+/-14.2 and 15.8+/-11.1 in the former, and 37.1+/ 7.8 and 8.7+/- 13.4 in the latter, respectively. Thus, pre-operative RCT enhanced apoptotic cell death and abated proliferative activity significantly (P<0.05), regardless of histological differentiation. Enhancement of apoptosis was more prominent in the group treated with FT or 5-FU than with BLM or PEP. Oral SCC with >20% of nuclear p53-positive tumor cells was noted in six cases. Enhanced TI and abadement of KI did not differ among the p53-positive and -negative tumors. PMID- 9736428 TI - Development of smokeless tobacco-induced oral mucosal lesions. AB - This study examined clinical and inflammatory mediator parameters during the development of snuff-induced mucosal lesions. Nineteen smokeless tobacco (ST) users placed moist snuff at designated new placement sites over either a 2- or 7 day period. By day 2, the predominant clinical alteration was an erythematous reaction, and one-third of the subjects demonstrated white striations in combination with erythema or ulceration. By 7 days, 56% of the subjects displayed white striated lesions. Tissue concentrations (pg/mg) of IL-1beta were 4.6+/-1.6 at new placement sites as compared with 0.7+/-0.4 at control sites (P<0.05). PGE2 and IL-1alpha concentrations also were significantly higher (P<0.05) at new placement sites as compared with control sites (9.4+/-2.2 vs 4.1+/-0.7 and 6.2+/ 1.3 vs 3.2+/-0.7, respectively). In view of the recognized role of PGE2 and IL-1 in immune and inflammatory functions, these mediators may play a role in the pathogenesis of clinical alterations at ST placement sites. PMID- 9736429 TI - Telomerase activity in oral lichen planus. AB - The purpose of this study was to determine the expression of telomerase in refractory oral lichen planus. Using a polymerase chain reaction-based telomerase activity assay, we investigated telomerase activity in 20 oral lichen planus specimens (erosive 9, atrophic 11). Telomerase activity was detected in 14 cases (erosive 7, atrophic 7). Furthermore, 13 cases of lichen planus with mild dysplasia proved telomerase-positive in eight specimens and six of seven cases devoid of dysplasia were also positive in the telomerase assay. The data indicate that, in general, telomerase activity might be frequently detectable in OLP. The data also suggest that telomerase activity might not be particularly associated with the premalignant phenotype in OLP. PMID- 9736430 TI - Localization of mRNA for inflammatory cytokines in radicular cyst tissue by in situ hybridization, and induction of inflammatory cytokines by human gingival fibroblasts in response to radicular cyst contents. AB - The expression of mRNA encoding the inflammatory cytokines interleukin-1alpha (IL 1alpha), interleukin-1beta (IL-1beta), interleukin-6 (IL-6), interleukin-8 (IL-8) and tumor necrosis factor alpha(TNF-alpha) have been examined in radicular cysts by in situ hybridization. Furthermore, the biological activity of the contents of radicular cysts (RCC) has been assayed by adding extracts of RCC to cultured human gingival fibroblasts (HGFs) and analyzing the culture medium for the release of inflammatory cytokines. In the epithelial layer, keratinocytes expressed all cytokine mRNAs examined at various levels. Basal layer cells expressed mRNA for each cytokine. In the subepithelial granulation tissue of the cysts, fibroblasts and macrophages expressed mRNA for IL-6, IL-8, IL-1beta and TNF-alpha mRNA at varying levels; especially clear expression of TNF-alpha and IL 1beta mRNA was detected on macrophages. The infiltrating lymphoid cells, largely composed of T cells and plasma cells, expressed these cytokine mRNAs, especially those encoding IL-6 and IL-8, at various levels. In vitro analysis indicated dose dependent release of both IL-6 and IL-8 by HGFs in response to RCC. After heating to 100 degrees C for 10 min, RCC almost completely failed to stimulate IL-6 release from HGFs. Furthermore, anti-IL-1beta antibody (neutralization test) did not prevent the stimulation of IL-6 release by RCC. Significant amounts of IL-6 and IL-8 were detected in RCC in two cases, and a trace amount of IL-1beta was detected in one case. This study demonstrated the wide expression of mRNA encoding inflammatory cytokines in radicular cyst tissues, and RCC itself was capable of stimulating IL-6 and IL-8 production from HGFs. PMID- 9736431 TI - High-resolution DNA flow cytometry in papillary cystadenoma lymphomatosum (Warthin's tumour). AB - Twenty-eight examples of papillary cystadenoma lymphomatosum (Warthin's tumour) of the parotid gland were analysed by high-resolution DNA flow cytometry. The mean coefficient of variation was found to be 1.19% (SD: 0.41). All tumours were DNA diploid. These results did not correspond with expected deviations based on published chromosomal studies. Also, the homogeneously low S-phase fractions (mean: 4.8%; SD: 2.7) found did not support the hypothesis of etiologically distinctive subgroups in these tumours. PMID- 9736432 TI - Palatal pleomorphic adenoma with florid squamous metaplasia: a potential diagnostic pitfall. AB - We report a case of pleomorphic adenoma with extensive squamous metaplasia in the palate of a 52-year-old Chinese man. The dimensions of the tumor were 15 x 13 x 13 mm. More than 95% of the epithelial element in the tumor was composed of sheets of squamous cells with cyst formation. The pathogenesis, potential diagnostic pitfalls, proliferative activity and cytokeratin expression of this tumor are discussed. PMID- 9736433 TI - Multiple supernumerary teeth in two brothers: a case report. AB - Supernumerary teeth (hyperdontia) are relatively common in the general population and occur more frequently in patients with a family history of such teeth. Supernumerary teeth have been reported in many genetic syndromes, but multiple supernumerary teeth occurring as an isolated non-syndromic trait are rare. This article describes a rare non-syndromic variety of multiple impacted supernumerary teeth in two brothers. PMID- 9736434 TI - The place of ACE inhibition in the modern therapeutic era: beyond blood pressure control. Introduction. PMID- 9736435 TI - Hypertensive patients and diabetes: a high-risk population. AB - Rising worldwide rates of diabetes mellitus heighten the need to maintain adequate metabolic control in diabetic patients and to control for other cardiovascular risk factors, such as lipid profile disturbances, high blood pressure, and smoking habits. This is especially the case in diabetic patients who also present with hypertension, a co-morbid state that is present in at least 50% of Type 1 and Type 2 diabetic patients. Cardiovascular disease is present in 75% of all diabetes-related deaths, and the concomitant condition of diabetes and hypertension is believed to act synergistically on elevating the risk for cardiovascular disease. A number of trials have demonstrated a greater incidence of cardiovascular disease end points in diabetic hypertensive patients than in diabetic normotensive patients. Furthermore, hypertension is associated not only with an increased risk for cardiovascular mortality but also for microvascular complications in patients with diabetes. Adequate treatment of high blood pressure is imperative in these patients. The effectiveness of antihypertensive treatment can be measured not only by the degree of reduction in blood pressure but also by assessment of the effects on urinary albumin excretion rate. It is assumed that the greater the reduction in urinary albumin excretion rate, the greater the renoprotective effect. Treatment choices should be evidence-based, i.e., physicians should concentrate not only on the treatment of hypertension but also on improving glycemic control and lipid profile disorders, when necessary. When viewed in this regard, angiotensin-converting enzyme inhibitors, low-dose diuretics, and in some cases beta-blockers, should be considered agents of choice in hypertensive diabetic patients. PMID- 9736436 TI - Relationships among diabetes, microalbuminuria, and ACE inhibition. AB - Increasing worldwide rates of diabetes mellitus, combined with the significant micro- and macrovascular complications that accompany the disease, point to a heightened need to develop simple, rational strategies to protect against end organ damage, particularly diabetic nephropathy. Although simple strategies do exist, i.e., early diagnosis of microalbuminuria (MAU) followed by nephroprotective therapy with angiotensin-converting enzyme (ACE) inhibitors, the use of these techniques should be increased. This is especially true for primary care physicians, who will continue to handle the majority of diabetic patients. To combat the underuse of this dual approach, this article reviews a stepwise approach to identifying early MAU so that therapeutic measures can be undertaken before the disorder progresses to diabetic nephropathy and, consequently, to end stage renal disease in the majority of patients. Once a diagnosis of MAU is made, a variety of trials have demonstrated that ACE inhibitor therapy should be considered the standard therapy to retard worsening albuminuria and subsequent renal disease. ACE inhibitors can retard the progression of microalbuminuria and can lower the percentage of patients who progress to end-stage renal disease and death. Significant data indicate that ACE inhibitors should be used in MAU diabetics regardless of the level of blood pressure. In particular, the ACE inhibitor fosinopril may offer advantages because of its dual route of elimination, thus simplifying dosing in renally impaired patients. The newly developed angiotensin II receptor antagonists should be considered in patients intolerant to ACE inhibitors because of the similar effect on the interruption of the renin-angiotensin system. PMID- 9736437 TI - New evidence on the prevention of cardiovascular events in hypertensive patients with type 2 diabetes. AB - Recent trials in hypertensive patients with type 2 diabetes reveal important differences in the risk for major cardiovascular events when individual agents are compared. In the Fosinopril Amlodipine Cardiovascular Events Trial (FACET), 380 patients with hypertension and type 2 diabetes were randomized to fosinopril or amlodipine and followed for up to 3.5 years to assess effects on serum lipids. Although both agents effectively controlled blood pressure, amlodipine caused a significantly greater decrease in systolic pressure. At the end of the trial, serum cholesterol, high-density lipoprotein cholesterol, triglycerides, HbA1c, serum glucose, plasma insulin, serum creatinine, and microalbuminuria were similar in both groups. The patients randomized to fosinopril were significantly less likely to experience the prospectively defined combined outcome of acute myocardial infarction (MI), hospitalized angina, or stroke compared to those randomized to amlodipine (RR 0.49; 95% CI 0.26-0.95). In the Appropriate Blood pressure Control in Diabetes (ABCD) trial, 470 patients with hypertension and type 2 diabetes who were randomized to long-acting nisoldipine had an adjusted sevenfold increased risk for acute MI compared to those randomized to enalapril (RR 7.0; 95% CI 2.3-21.4). In the Multicenter Isradipine Diuretic Atherosclerosis Study (MIDAS) trial, the patients with hypertension and above the median of HbA1c (> or =6.7%) randomized to isradipine had a threefold increased risk for major cardiovascular events compared to those randomized to hydrochlorothiazide (RR 2.81; 95% CI 1.09-7.26). These findings are supported by several observational studies. Therefore, evidence is emerging that angiotensin-converting enzyme inhibitors and low-dose diuretics may be more effective than calcium antagonists for prevention of cardiovascular events in hypertensive patients with diabetes or impaired glucose control. PMID- 9736438 TI - Evidence-based medicine and ACE inhibition. AB - The use of angiotensin-converting enzyme (ACE) inhibitors has been generally beneficial in the treatment of many clinical conditions characterized by a significant degree of cardiovascular and renal involvement. Most of the available data on the benefits of ACE inhibitors have come from well-conducted large clinical trials that have provided much information supporting the use of ACE inhibitors, in agreement with the basic principles of evidence-based medicine. In particular, ACE inhibitors improve blood pressure control in patients with hypertension and have proved to be beneficial in patients with left ventricular (LV) systolic dysfunction and chronic congestive heart failure (CHF). Improved survival rates after the use of ACE inhibitors have been also demonstrated in patients with acute myocardial infarction (MI), whether or not the condition is complicated by acute CHF. More recently, some studies have demonstrated the ability of ACE inhibitors (particularly fosinopril) to prevent the long-term development of CHF in patients treated acutely during MI and without baseline LV dysfunction. ACE inhibitors appear to improve the long-term prognosis of patients with coronary artery disease (CAD) and to reduce the occurrence of re-infarction, as demonstrated in the Studies of Left Ventricular Dysfunction (SOLVD) trial and the Survival and Ventricular Enlargement study (SAVE). Finally, a protective role for ACE inhibitors has been reported even in diabetic hypertensive patients, in whom such agents can significantly reduce the occurrence of major cardiovascular events (CAD and stroke) with a pattern that is largely independent of blood pressure control and is not observed with the use of calcium antagonists. These data confirm the strong involvement of the renin-angiotensin system in the pathophysiology of vascular diseases and strongly support the role of ACE inhibitors as drugs for present and future therapy. PMID- 9736439 TI - Neurohormonal markers of clinical outcome in cardiovascular disease: is endothelin the best one? AB - Endothelin-1 (ET-1) is the most potent vasoconstrictor yet described. The active 21-amino-acid peptide is derived from the conversion of the inactive precursor "Big ET-1" by an enzyme called endothelin-converting enzyme. In addition to its potent action as a vasoconstrictor, endothelin promotes growth and proliferation of smooth muscle and myocardial hypertrophy. ET-1 levels are elevated in acute myocardial infarction (MI), atherosclerosis, renal failure, diabetes, pulmonary hypertension, and congestive heart failure (CHF). ET-1 levels correlate extremely well with the seriousness of the pathophysiologic condition. ET-1 levels at 72 h post MI accurately predict long-term survival. In patients with heart failure, ET 1 levels also predict long-term outcome, with the prognosis being severely compromised in patients with elevated ET-1 levels. Levels of plasma big ET-1 have been demonstrated to predict 1-year mortality and have been shown to be a better predictor of 1-year outcome than plasma atrial natriuretic peptide and norepinephrine, NYHA class, age, and echocardiographic left ventricular parameters. Although a small number of studies have reported beneficial effects of ACE inhibitors on ET-1 levels in animal models, most reports in humans have not found an effect of ACE inhibitors on ET-1 levels. Only one ACE inhibitor, fosinopril, has been shown to be effective in normalizing ET-1 levels in clinically relevant situations, such as the long-term study of patients with CHF. This observation may point to a superior role of fosinopril compared with other ACE inhibitors in CHF patients and may indicate beneficial effects of fosinopril beyond blood pressure control. PMID- 9736440 TI - Immunoresponsiveness in chronic hepatitis C patients: correlation between tissue and serum findings. AB - In the present study, intrahepatic CD8+ lymphocyte infiltrates as well as HLA class I and CD54 (ICAM-1) antigen expression at both tissue and serum levels were evaluated in 54 untreated patients with chronic hepatitis C stratified on the basis of histological diagnosis (Chronic Persistent Hepatitis/Chronic Lobular Hepatitis -CPH/CLH- and Chronic Active Hepatitis -CAH-: 22 and 32 subjects, respectively). The relationships between soluble HLA-I (sHLA-I) and ICAM-1 (sICAM 1) serum levels and their membrane-bound counterparts, CD8+ liver infiltration and serum alanine aminotransferase (ALT) were also studied. A strong HLA-I and CD54 tissue expression, associated to the presence of CD8+ cell infiltrates in necro-inflammatory areas, and elevated sHLA-I and sICAM-1 serum amounts were observed in all patients. At the same time, no difference was found at tissue level between the two groups of patients with respect to the mean scores of HLA-I and CD54 expression, while CAH subjects displayed a significantly higher CD8 periportal and lobular reactivity in comparison to the other subset. Serological assays outlined higher values of circulating HLA-I molecules in CPH/CLH patients and higher sICAM-1 levels in the CAH group. Finally, a negative correlation was found between sHLA-I and ALT in CAH subjects while, in all patients, sICAM-1 positively correlated with both CD8 tissue infiltration and ALT. Our findings confirm the occurrence of an immune activation status during chronic hepatitis C and suggest that sHLA-I molecules might play a down-modulating role on immunoresponsiveness of these patients. PMID- 9736441 TI - Mistletoe lectin I-induced effects on human cytotoxic lymphocytes. I. Synergism with IL-2 in the induction of enhanced LAK cytotoxicity. AB - This report demonstrates that in vitro activation of human cells with the beta galactoside-specific lectin from mistletoe (ML-I) or interleukin-2 (IL-2) results in different patterns of activation and function of cytotoxic cells. It is now well established that natural killer (NK) and lymphokine-activated killer (LAK) cytotoxicity is mainly mediated by resting (NK) and IL-2-activated (LAK) CD56 positive (+) cells respectively. Culture of peripheral blood lymphocytes (PBL) for 3 days with ML-I led to expansion and activation of T cells which demonstrated NK- and LAK-like cytotoxicity. T lymphocyte subset analysis revealed that in total PBL, ML-I preferentially stimulated and expanded CD8+ T cells which mediated the cytotoxic effect. Incubation of highly purified CD8+ T cells alone with ML-I did not lead to induction of cytotoxicity, which required the presence of both CD4+ and CD14+ (monocytes) cells, suggesting that ML-I does not exert a direct effect on CD8+ T cells. Activation of PBL with both ML-I and IL-2 resulted in simultaneous induction of T and CD56+ cell-mediated NK and LAK cytotoxicity. These data suggest that treatment with ML-I and IL-2 might provide an approach to induce maximum cytotoxicity against tumors and to recruit both T and NK cells for tumor therapy. PMID- 9736442 TI - Profile of cerebrospinal fluid and serum cytokines in patients with relapsing remitting multiple sclerosis: a correlation with clinical activity. AB - Levels of tumor necrosis factor (TNF)-alpha, granulocyte macrophage-colony stimulating factor (GM-CSF), interleukin (IL)-10 and transforming growth factor (TGF)-beta in cerebrospinal fluid (CSF) and serum of 29 patients with multiple sclerosis (MS) of the relapsing-remitting type and of 20 controls with other non inflammatory neurological diseases were studied. Sixteen patients were in the active phase of disease and 13 in remission. In CSF, higher IL-10 and TGF-beta concentrations were found in patients with a stable phase of MS, while in the active phase there were elevated levels of TNF-alpha and GM-CSF. These results suggest that a different cytokine pattern could be probably involved in the pathogenesis of relapsing-remitting MS. PMID- 9736443 TI - Modulation of eosinophilic chemotaxis with azelastine and budesonide in allergic patients. AB - In this paper, the effect of azelastine hydrochloride, a potent inhibitor of leukotrienes (LTs) and H1 receptors for histamine, was assessed as regards modulation of in vitro eosinophilic chemotaxis. In this respect, chemotaxis of eosinophils (EOS), isolated from the peripheral blood of untreated allergic subjects in the acute phase, was significantly diminished after in vitro treatment with azelastine in comparison to values before treatment. When EOS were pre-incubated with serial dilutions of the drug, it was observed that azelastine inhibited chemotaxis in a dose-dependent fashion. Since azelastine acts in vitro as a regulator of the calcium pump, EOS were pre-incubated with different concentrations (0.6 and 3.0 mM) of Ca++. In these experimental conditions azelastine was able to reduce EOS chemotactic activity only in the presence of 0.6 mM Ca++, whereas with higher Ca++ concentrations (3.0 mM) the inhibitory effect of the drug was abrogated. On the other hand, particular attention was paid to inhaled budesonide, a non halogenated glucocorticosteroid derivative, structurally related to 16 alpha-hydroxy prednisolone, which represents a helpful for treatment mild to moderate asthma. Data obtained after in vitro treatment with budesonide of a group of allergic patients demonstrated that EOS chemotactic activity was significantly reduced in these subjects. Conclusively our data show that 1) azelastine acts as a dose-dependent antagonist of chemotaxis; 2) it may exert this action by inhibiting Ca++ flow into cells; 3) inhaled budesonide may induce inhibition of bronchial inflammation by downregulating EOS chemotactic capacity. PMID- 9736444 TI - Sulfide enhancement of PMN apoptosis. AB - Hydrogen sulfide is a toxic metabolite released by several bacterial agents under anaerobic conditions. In the present paper, we investigated the effects of sulfide on polymorphonuclear cell (PMN) apoptosis, a mechanism suggested for limiting the toxic potential of neutrophils in inflammatory sites. We showed that 1 mM sulfide (concentration not conditioning PMN viability) is able to enhance the apoptotic fate of human granulocytes by increasing: i) the number of cells containing pyknotic nuclei, ii) the internucleosomal cleavage, and, iii) the intensity of tubulin immunofluorescence staining. The sulfide effect is partially prevented by ionomycin and this finding is consistent with the hypothesis of the inhibiting role played by high levels of cytosolic calcium in PMN apoptosis modulating. PMID- 9736445 TI - The porphyrin photosensitizer Photofrin elevates murine splenic erythropoiesis. AB - Changes occurring within the spleens of the genetically distinct DBA/1 and DBA/2 mouse strains produced by the photosensitizer Photofrin in the absence of direct light exposure were analyzed. Photofrin significantly increased spleen weight, cellularity and erythroid progenitor levels in both mouse strains tested. The expression of heat stable antigen (HSA), a marker present upon different immature leukocytes as well as certain fully-differentiated cell types including erythrocytes, was increased in the spleens of mice given Photofrin. It was shown for Photofrin-injected DBA/1 mice that a spleen cell population which expressed high levels of HSA also bound the iron transport protein transferrin. Photofrin increases the splenic demand for iron by promoting erythropoietic activity within the tissue. PMID- 9736446 TI - Protective effect of oral administration of a traditional medicine, Juzen-Taiho To, and its components on lethal Candida albicans infection in immunosuppressed mice. AB - Protective effects of a kampo medicine, Juzen-taiho-to (TJ-48) and its herbal components against experimental candidiasis in cyclophosphamide-induced immunosuppressive mice were investigated. ICR mice were immunosuppressed by intraperitoneal treatment with cyclophosphamide (day-4) and were orally given TJ 48 or one of its 10 herbal components for 4 consecutive days (day-4--1). They were then challenged intravenously with a lethal dose of Candida albicans (day 0). An oral dose of 1 g/kg/day of TJ-48 prolonged their life span. A similar protective effect was obtained by treatment with its component drugs Ginseng radix, Glycyrrhizae radix, Atractylodis lancea rhizoma or Cnidii rhizoma. These herbal components were suggested to have a main role in the protective effect of Juzen-taiho-to against Candida infection. PMID- 9736447 TI - Endothelin-1 and NO2/NO3 circulating levels after short-term (1h) oxygen supplementation in patients with chronic respiratory failure during long-term oxygen treatment (LTOT). AB - The effect of acute oxygen administration on endothelin-1 (ET-1) and nitrates (NO.2/NO.3), the latter as stable end products of nitric oxide (NO), were evaluated in arterial and venous blood of chronic respiratory failure (CRF) patients underwent to a continuous long-term oxygen therapy (LTOT). After one hour of oxygen supplementation, ET-1 showed a marked and significant decrease more pronounced in venous blood whereas no statistical change in NO.2/NO.3 concentrations were observed in both arterial and venous blood. There are evidences for increased expression of ET-1 in several pulmonary diseases and for ET-1 plasma reduction in Adult Respiratory Distress Syndrome (ARDS) in patients which recovered. ET-1 is a potent human pulmonary vessel constrictor and may have other effects including plasma exudation, increased mucus secretion and a increased fibrinogenesis. Our data suggest that the improvement in air function, evaluated in part by the decreased release of inflammatory mediators and mainly by reduction in the pulmonary arterial resistance, may be a consequence of the decrease in ET-1 content in the lungs of CRF patients treated with LTOT. PMID- 9736448 TI - Polymorphonuclear phagocytosis and killing in workers occupationally exposed to hexachlorobenzene. AB - Phagocytosis and intracellular killing of Candida albicans and Candida pseudotropicalis by neutrophils from 66 workers exposed to chlorinated compounds were studied. Phagocytosis with both antigens was normal in all the workers studied. However, lytic activity of the neutrophils in the presence of both antigens, C. albicans and C. pseudotropicalis was impaired. These findings might be related to a derangement in the antioxidant mechanisms and to some interference with GSH levels that might lead to the abnormal functions observed in neutrophils. PMID- 9736449 TI - Dual cytoplasmic and nuclear distribution of the novel arsenite-stimulated human ATPase (hASNA-I). AB - The arsenite-stimulated human ATPase (hASNA-I) protein is a distinct human ATPase whose cDNA was cloned by sequence homology to the Escherichia coli ATPase arsA. Its subcellular localization in human malignant melanoma T289 cells was examined to gain insight into the role of hASNA-I in the physiology of human cells. Immunocytochemical staining using the specific anti-hASNA-I monoclonal antibody 5G8 showed a cytoplasmic, perinuclear, and nucleolar distribution. Subcellular fractionation indicated that the cytoplasmic hASNA-I was soluble and that the perinuclear distribution was due to association with the nuclear membrane rather than with the endoplasmic reticulum. Its presence in the nucleolus was confirmed by showing colocalization with an antibody of known nucleolar specificity. Further immunocytochemical analysis showed that the hASNA-I at the nuclear membrane was associated with invaginations into the nucleus in interphase cells. These results indicate that hASNA-I is a paralogue of the bacterial ArsA protein and suggest that it plays a role in the nucleocytoplasmic transport of a nucleolar component. PMID- 9736450 TI - Prereplicative increase of nuclear matrix-bound DNA polymerase-alpha and primase activities in HeLa S3 cells following dilution of long-term cultures. AB - We investigated the association of DNA polymerase and DNA primase activity with the nuclear matrix in HeLa S3 cells diluted with fresh medium after having been cultured without any medium change for 7 days. Flow cytometric analysis demonstrated that just before dilution about 85% of the cells were in the G1 phase of the cycle, whereas 8% were in the S phase. After dilution with fresh medium, 18-22 h were required for the cell population to attain a stable distribution with respect to the cell cycle. At that time, about 38% of the cells were in the S phase. DNA polymerase and DNA primase activity associated with the nuclear matrix prepared from cells just before dilution represented about 10% of nuclear activity. As judged by [3H]-thymidine incorporation and flow cytometric analysis, an increase in the number of S-phase cells was evident at least 6 h after dilution. However, as early as 2 h after dilution into fresh medium, a striking prereplicative increase of the two activities was seen in the nuclear matrix fraction but not in cytosol or isolated nuclei. Both DNA polymerase and primase activities bound to the matrix were about 60% of nuclear activity. Overall, the nuclear matrix was the cell fraction where the highest induction (about 10-fold) of both enzymatic activities was seen at 30 h after dilution, whereas in cytosol and isolated nuclei the increase was about two- and fourfold, respectively. Typical immunofluorescent patterns given by an antibody to 5 bromodeoxyuridine were seen after dilution. These findings, which are at variance with our own previous results obtained with cell cultures synchronized by either a double thymidine block or aphidicolin exposure, strengthen the contention that DNA replication is associated with an underlying nuclear structure and demonstrate the artifacts that may be generated by procedures commonly used to synchronize cell cultures. PMID- 9736451 TI - Involvement of heat shock elements and basal transcription elements in the differential induction of the 70-kDa heat shock protein and its cognate by cadmium chloride in 9L rat brain tumor cells. AB - Exposure of 9L rat brain tumor cells to 40-100 microM CdCl2 for 2 h leads to an induction of a wide spectrum of heat shock proteins (HSPs). We have demonstrated that induction of the 70-kDa HSP (HSP70) and enhanced expression of its cognate (HSC70) by cadmium are concentration dependent and that the induction kinetics of these HSP70s are different. The increased synthesis of the HSP70s is accompanied by the increase in hsp70 and hsc70 mRNA levels, indicative of transcriptional regulation of the heat shock genes. Electrophoretic mobility shift assay (EMSA) using probes encompassing heat shock element (HSE), TATA, GC, and CCAAT boxes derived from the promoter regions of the heat shock genes shows distinguished binding patterns between hsp70 and hsc70 genes in both control and cadmium treated cells. The results indicate that, in addition to the HSEs, the basal transcription elements are important in the regulation of the heat shock genes. The binding patterns of the corresponding transcription factors of these elements are examined by EMSA by using extended promoter fragments from respective heat shock genes with sequential addition of excess oligonucleotides encompassing individual transcription elements. Taken together, our results show that the differential induction of hsp70 and hsc70 involves multiple transcription factors that interact with HSE, TATA, GC, and CCAAT boxes. PMID- 9736452 TI - Chemokine receptor CCR5 functionally couples to inhibitory G proteins and undergoes desensitization. AB - Chemokine receptor CCR5 is not only essential for chemotaxis of leukocytes but also has been shown to be a key coreceptor for HIV-1 infection. In the present study, hemagglutinin epitope-tagged human CCR5 receptor was stably expressed in Chinese hamster ovary cells or transiently expressed in NG108-15 cells to investigate CCR5-mediated signaling events. The surface expression of CCR5 was confirmed by flow cytometry analysis. The CCR5 agonist RANTES stimulated [35S]GTPgammaS binding to the cell membranes and induced inhibition on adenylyl cyclase activity in cells expressing CCR5. The effects of RANTES were CCR5 dependent and could be blocked by pertussis toxin. Furthermore, overexpression of Gialpha2 strongly increased both RANTES-dependent G-protein activation and inhibition on adenylyl cyclase in cells cotransfected with CCR5. These data demonstrated directly that activation of CCR5 stimulated membrane-associated inhibitory G proteins and indicated that CCR5 could functionally couple to G protein subtype Gialpha2. The abilities of CCR5 to activate G protein and to inhibit cellular cAMP accumulation were significantly diminished after a brief prechallenge with RANTES, showing rapid desensitization of the receptor-mediated responsiveness. Prolonged exposure of the cells to RANTES caused significant reduction of surface CCR5 as measured by flow cytometry, indicative of agonist dependent receptor internalization. Our data thus demonstrated that CCR5 functionally couples to membrane-associated inhibitory G proteins and undergoes agonist-dependent desensitization and internalization. PMID- 9736453 TI - Molecular cloning of a mutated HOXB7 cDNA encoding a truncated transactivating homeodomain-containing protein. AB - Homeodomain-containing proteins regulate, as transcription factors, the coordinated expression of genes involved in development, differentiation, and malignant transformation. We report here the molecular cloning of a mutated HOXB7 transcript encoding a truncated homeodomain-containing protein in MCF7 cells. This is a new example of mutation affecting the coding region of a HOX gene. In addition, we detected two HOXB7 transcripts in several breast cell lines and demonstrated that both normal and mutated alleles were expressed at the RNA level in MCF7 cells as well as in a variety of breast tissues and lymphocytes, suggesting that a truncated HOXB7 protein might be expressed in vivo. Using transient co-transfection experiments, we demonstrated that both HOXB7 proteins can activate transcription from a consensus HOX binding sequence in breast cancer cells. Our results provide evidence that HOXB7 protein has transcription factor activity in vivo and that the two last amino acids do not contribute to this property. PMID- 9736454 TI - Osteoblastic phenotype of rat marrow stromal cells cultured in the presence of dexamethasone, beta-glycerolphosphate, and L-ascorbic acid. AB - We investigated the effects of the time course of addition of osteogenic supplements dexamethasone, beta-glycerolphosphate, and L-ascorbic acid to rat marrow stromal cells, and the exposure time on the proliferation and differentiation of the cells. It was the goal of these experiments to determine the time point for supplement addition to optimize marrow stromal cell proliferation and osteoblastic differentiation. To determine this, two studies were performed; one study was based on the age of the cells from harvest, and the other study was based on the duration of exposure to supplemented medium. Cells were seen to proliferate rapidly at early time points in the presence and absence of osteogenic supplements as determined by 3H-thymidine incorporation into the DNA of replicating cells. These results were supported by cell counts ascertained through total DNA analysis. Alkaline phosphatase (ALP) activity and osteocalcin production at 21 days were highest for both experimental designs when the cells were exposed to supplemented medium immediately upon harvest. The ALP levels at 21 days were six times greater for cells maintained in supplements throughout than for control cells cultured in the absence of supplements for both studies, reaching an absolute value of 75 x 10(-7) micromole/min/cell. Osteocalcin production reached 20 x 10(-6) ng/cell at 21 days in both studies for cells maintained in supplemented medium throughout the study, whereas the control cells produced an insignificant amount of osteocalcin. These results suggest that the addition of osteogenic supplements to marrow-derived cells early in the culture period did not inhibit proliferation and greatly enhanced the osteoblastic phenotype of cells in a rat model. PMID- 9736455 TI - Beta1 integrin cytoplasmic domain regulates the constitutive conformation detected by MAb 15/7, but not the ligand-induced conformation. AB - The anti-integrin beta1 MAb 15/7 sometimes may be a reporter of integrin activation or ligand occupancy. However, certain beta1 tail deletions eliminate ligand binding despite inducing maximal constitutive 15/7 expression [Puzon Mclaughlin et al. (1 996): J Biol Chem 271:16580-16585]. Here we describe beta1 tail mutations (e.g., double point mutations [D759L/F763L, F766L/E769L], or replacement of the beta1 tail by the beta5 tail) that prevent rather than induce constitutive appearance of the 15/7 epitope. Despite variable losses of constitutive 15/7 epitope, these mutants all retained a similar inducible 15/7 epitope component as seen upon incubation with GRGDSP peptide ligand. In addition, constitutive 15/7 expression did not correlate with integrin localization into focal adhesions. In conclusion, we show for the first time for a fully functional integrin that specific mutations within the beta1 tail can down-regulate the constitutive appearance of an extracellular conformation defined by MAb 15/7. Because this regulation occurs away from the ligand binding site and does not correlate with responsiveness to integrin ligand, cell adhesion, or localization into focal adhesions, a novel type of conformational regulation is suggested. PMID- 9736457 TI - Role of M-line proteins in sarcomeric titin assembly during cardiac myofibrillogenesis. AB - A rat polyclonal anti-M-line protein antiserum and three mouse monoclonal anti titin antibodies (E2, F3, and A12) were used to study the spatiotemporal relationship between M-line proteins and titin during myofibril assembly in cultured chicken cardiomyocytes by immunofluorescence microscopy. In day 2 cultures, M-line proteins and titin were detected as punctate staining in most cardiomyocytes, which possessed many nonstriated fibrils. At a late stage (day 3 cultures), M-line proteins were incorporated into dot-like structures along nonstriated fibrils, while titin staining was continuous on these structures. As development progressed, M-line proteins were registered in periodic pattern in the mid-A band. In cardiomyocytes from day 5 cultures, the titin bands were separated by an unstained region, and achieved their adult doublet pattern. Thus, the organization of titin in the sarcomere appears to occur later than that of M line proteins in the M-line. Our morphological data indicate that the early registration of M-line proteins in primitive myofibrils may guide titin filament alignment via interaction between M-line proteins and titin. In order to investigate the role of M-line proteins in the assembly of titin filaments, anti M-line protein or anti-titin antibodies were introduced into cultured cardiomyocytes by electroporation to functionally bind the respective proteins, and the profile of myofibril assembly was examined. Cardiomyocytes from day 2-3 cultures with incorporated anti-M-line protein antibodies became shrunk, and exhibited defective myofibrillar assembly, as shown by the failure of titin to assemble into a typical sarcomeric pattern. Incorporation of anti-titin antibody E2, which recognizes the M-line end domain of titin, resulted in the failure of M line proteins organized into the M-line structure, as shown by random, sporadic staining with anti-M-line protein antibody. These studies confirm the essential role of M-line proteins in the organization of titin filaments in the sarcomere and that the interaction between titin and M-line proteins is crucial to the formation of the M-line structure. PMID- 9736456 TI - Arachidonic acid metabolism by adult human osteoblast-like cells exhibits sexually dimorphic characteristics. AB - The eicosanoids, including prostaglandin E2 (PGE2) and other bioactive arachidonic acid metabolites, are important local mediators of bone remodeling. Presumably, the limited or excessive synthesis of the eicosanoids could compromise bone homeostasis. We have noted that the stimulated release of arachidonic acid by adult male donor derived human osteoblast-like (hOB) cells exceeded the stimulated release measured for female-derived hOB cells by 1.5 fold. Assays of PGE2 biosynthesis by cytokine-stimulated hOB cells also demonstrated a sex-linked difference, such that male hOB cell PGE2 production exceeded female cell production by 1.6-2.2-fold. The calcium-dependent cytoplasmic phospholipase A2 activity in subcellular fractions prepared from hOB cell homogenates was higher in both the cytosolic (1.6-fold) and particulate (1.5 fold) fractions from the male cells than in those prepared from female hOB cells, suggesting a molecular basis for the observed sexually dimorphic characteristics related to arachidonic acid metabolism by hOB cells. The relatively limited capacity of the female cells may limit needed intracellular and intercellular signaling during bone remodeling, thereby contributing to the development of bone pathology. PMID- 9736458 TI - Effects of gonadal and adrenal androgens in a novel androgen-responsive human osteoblastic cell line. AB - While androgens have important skeletal effects, the mechanism(s) of androgen action on bone remain unclear. Current osteoblast models to study androgen effects have several limitations, including the presence of heterogeneous cell populations. In this study, we examined the effects of androgens on the proliferation and differentiation of a novel human fetal osteoblastic cell line (hFOB/AR-6) that expresses a mature osteoblast phenotype and a physiological number (approximately 4,000/nucleus) of androgen receptors (AR). Treatment with 5alpha-dihydrotestosterone (5alpha-DHT) inhibited the proliferation of hFOB/AR-6 cells in a dose-dependent fashion, while it had no effect on the proliferation of hFOB cells, which express low levels of AR (<200/nucleus). In hFOB/AR-6 cells, co treatment with the specific AR antagonist, hydroxyflutamide abolished 5alpha-DHT induced growth inhibition. Steady-state levels of transforming growth factor beta1 (TGF-beta1) and TGF-beta-induced early gene (TIEG) mRNA decreased after treatment of hFOB/AR-6 cells with 5alpha-DHT, suggesting a role for the TGF-beta1 TIEG pathway in mediating 5alpha-DHT-induced growth inhibition of hFOB/AR-6 cells. In support of this, co-treatment of hFOB/AR-6 cells with TGF-beta1 (40 pg/ml) reversed the 5alpha-DHT-induced growth inhibition, whereas TGF-beta1 alone at this dose had no effect on hFOB/AR-6 cell proliferation. Furthermore, treatment of hFOB/AR-6 cells with 5alpha-DHT and testosterone (10(-8) M) inhibited basal and 1,25-(OH)2D3-induced alkaline phosphatase (ALP) activity and type I collagen synthesis without affecting osteocalcin production. Thus, in this fetal osteoblast cell line expressing a physiological number of AR, androgens decrease proliferation and the expression of markers associated with osteoblast differentiation. These studies suggest that the potential anabolic effect of androgens on bone may not be mediated at the level of the mature osteoblast. PMID- 9736459 TI - Shear stress down-regulates gene transcription and production of adrenomedullin in human aortic endothelial cells. AB - Vascular endothelial cells are potent modulators of vascular tone in response to shear stress. Levels of vasoactive peptides such as adrenomedullin (AM), endothelin-1 (ET-1), C-type natriuretic peptide (CNP), and nitric oxide (NO) are affected by fluid shear stress. AM, a potent vasodilator and suppressor of smooth muscle cell proliferation, contains the shear stress responsive element (SSRE) "GAGACC" in its promoter region. To examine the role of AM in the shear stress response, cultured human aortic endothelial cells (HAoECs) were exposed to fluid shear stresses of 12 and 24 dynes/cm2 in a cone-plate shear stress loading apparatus for various time periods, and the levels of AM gene expression and peptide secretion from HAoECs were measured by Northern blotting analysis and radioimmunoassay (RIA), respectively. Both AM gene transcription and AM peptide levels were down-regulated by fluid shear stress in a time- and magnitude dependent manner. Our results demonstrate that the normal level of arterial shear stress down-regulates AM expression in HAoECs, suggesting that AM participates in the modulation of vascular tone by fluid shear stress. PMID- 9736460 TI - Opposing effects of cyclosporin A and tyrphostin AG-1478 indicate a role for Src protein in the cellular control of mineralization. AB - Cyclosporin A (CsA) induces osteoporosis but not through direct activation of osteoclasts. CsA also inhibits cell-mediated mineralization in marrow stromal cell culture, whereas the tyrphostin AG-1478 increases mineralization. These antagonistic effects on mineralization were used to discern molecules that underwent phosphorylation changes in association with their opposing effects on mineralization. In parallel, quantitative changes in Src protein were followed. Multiple dexamethasone (DEX)-stimulated stromal cell cultures were grown with and without a mineralization-inhibiting dose (0.1 microM) of CsA and were harvested on different days of DEX stimulation. Immunoblots of gel-fractionated cell extracts showed that the most noticeable changes in tyrosine phosphorylated proteins (TPP) were seen on day 8 of DEX stimulation. At least 15 TPP bands, mostly smaller than 53 kDa, were more prominent in CsA-treated cultures on day 8. Under CsA, Src protein quantity decreased on day 8, but its cleavage product (52/54 kDa) was sixfold more abundant then on day 7. Day 8 was chosen to test the effect of AG-1478 on the CsA-induced TPP changes. Dimethyl sulfoxide (DMSO) alone, the solvent of AG-1478, increased mineralization in CsA-treated versus CsA untreated cultures and slightly decreased Src and its cleavage product. AG-1478 at 5 microM, in CsA cultures increased the specific alkaline phosphatase activity threefold, with a slight change in mineralization relative to controls grown with DMSO alone. This was accompanied by decreased intensity of several TPP bands smaller than 36 kDa. In contrast, treatment with 50 microM of AG-1478 increased the intensity of TPP bands at the same molecular size range. This high AG-1478 dose decreased cell counts selecting mineralizing cells. The results indicate that increased Src protein cleavage product on day 8 by CsA is associated with mineralization inhibition, which is opposed by DMSO and 50-microM AG-1478, thus antagonizing the effect of CsA on mineralization. Direct or indirect interaction between Src and TPP, antagonistically affected by CsA and AG-1478, is likely to underlay cellular control of mineralization. Changes in p19 and p29 intensity showed association with mineralization that was reflected by a significant direct and inverse correlation, respectively, with calcium precipitation per cell. PMID- 9736461 TI - Several novel transcripts of glyceraldehyde-3-phosphate dehydrogenase expressed in adult chicken testis. AB - Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), in addition to being a classic glycolytic enzyme, is a multifunctional protein involved in relevant cell functions such as DNA replication, DNA repair, translational control of gene expression, and apoptosis. Although the multifunctional nature of GAPDH suggests versatility in the mechanisms regulating its expression, no major qualitative changes and few quantitative changes in the GAPDH transcripts have been reported. While studying the expression of GAPDH during spermatogenesis, we detected alternative initiations to TATA box and alternative splicings in the 5' region of the pre-mRNA, resulting in at least six different types of mRNAs. The amount and the polyadenylation of the GAPDH transcripts increased in mature testis in relation to immature testis and further increased when cell suspensions from mature testis were exposed to heat shock. These results suggest that alternative initiation, alternative splicing, and polyadenylation could provide the necessary versatility to the regulation of the expression of this multifunctional protein during spermatogenesis. PMID- 9736462 TI - Inhibition of Ca2+/calmodulin-dependent phosphatase activity by regucalcin in rat liver cytosol: involvement of calmodulin binding. AB - The regulatory effect of regucalcin on Ca2+/calmodulin-dependent phosphatase activity and the binding of regucalcin to calmodulin was investigated. Phosphatase activity toward phosphotyrosine, phosphoserine, and phosphothreonine in rat liver cytosol was significantly increased by the addition of Ca2+ (100 microM) and calmodulin (0.30 microM). These increases were clearly inhibited by the addition of regucalcin (0.50-1.0 microM) into the enzyme reaction mixture. The cytosolic phosphoamino acid phosphatase activity was significantly elevated by the presence of anti-regucalcin monoclonal antibody (0.2 microg/ml), suggesting that endogenous regucalcin in the cytosol has an inhibitory effect on the enzyme activity. This elevation was prevented by the addition of regucalcin (0.50 microM). Purified calcineurin phosphatase activity was significantly increased by the addition of calmodulin (0.12 microM) in the presence of Ca2+ (1 and 10 microM). This increase was completely inhibited by the presence of regucalcin (0.12 microM). The inhibitory effect of regucalcin was reversed by the addition of calmodulin with the higher concentration (0.36 microM). Regucalcin has been demonstrated to bind on calmodulin-agarose beads by analysis with sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The present study demonstrates that regucalcin inhibits Ca2+/calmodulin-dependent protein phosphatase activity in rat liver cytosol, and that regucalcin can bind to calmodulin. PMID- 9736463 TI - Is transcranial magnetic stimulation coming of age? PMID- 9736464 TI - Transcranial magnetic stimulation in study of the visual pathway. AB - The authors critically reviewed experiments in which transcranial magnetic stimulation (TMS) and repetitive TMS (rTMS) of the higher visual pathway were used. Topics include basic mechanisms of neural excitation by TMS and their relevance to the visual pathway (excitatory and inhibitory effects), TMS and rTMS of calcarine cortex (suppression, unmasking, and phosphenes), TMS of V5 (suppression), TMS and rTMS of higher level temporoparietooccipital areas (perceptual errors, unmasking, and inattention), the role of frontal lobe output in visual perception, and vocalization of perceived visual stimuli (role of consciousness of linguistic symbols). PMID- 9736465 TI - Studies of neuroplasticity with transcranial magnetic stimulation. AB - In recent years, there has been increasing interest in studies of brain plasticity. Although still loosely defined, this term describes the ability of the brain to change. Cortical plasticity encompasses a wide variety of phenomena and mechanisms, including modifications in cortical properties such as strength of internal connections, representational patterns, or neuronal modifications, either morphological or functional (Donoghue et al., 1996). We focus on the description of different ways in which transcranial magnetic stimulation (TMS) can be used to study patterns of reorganization and some of the mechanisms involved in these changes. Correlation between TMS and neuroimaging studies in humans and animal studies addressing similar questions is discussed. It is important to identify in each situation whether plasticity plays a beneficial role or is maladaptive in terms of functional compensation. The understanding of patterns, mechanisms, and functional relevance of cortical plasticity will hopefully lead to the design of effective strategies to enhance plasticity when it is beneficial and to down-regulate it when it is maladaptive. An example of a possible strategy, using TMS, is discussed. PMID- 9736466 TI - Transcranial magnetic stimulation: language function. AB - Studies of language using transcranial magnetic stimulation (TMS) have focused both on identification of language areas and on elucidation of function. TMS may result in either inhibition or facilitation of language processes and may operate directly at a presumptive site of language cortex or indirectly through intracortical networks. TMS has been used to create reversible "temporary lesions," similar to those produced by Wada tests and direct cortical electrical stimulation, in cerebral cortical areas subserving language function. Rapid-rate TMS over the left inferior frontal region blocks speech output in most subjects. However, the results are not those predicted from classic models of language organization. Speech arrest is obtained most easily over facial motor cortex, and true aphasia is rare, whereas right hemisphere or bilateral lateralization is unexpectedly prominent. A clinical role for these techniques is not yet fully established. Interfering with language comprehension and verbal memory is currently more difficult than blocking speech output, but numerous TMS studies have demonstrated facilitation of language-related tasks, including oral word association, story recall, digit span, and picture naming. Conversely, speech output also facilitates motor responses to TMS in the dominant hemisphere. Such new and often-unexpected findings may provide important insights into the organization of language. PMID- 9736467 TI - Study and modulation of human cortical excitability with transcranial magnetic stimulation. AB - Transcranial magnetic stimulation (TMS) can be applied in different paradigms to obtain a measure of various aspects of cortical excitability. These different TMS paradigms provide information about different neurotransmitter systems, enhance our understanding about the pathophysiology of neuropsychiatric conditions, and in the future may be helpful as a guide for pharmacological interventions. In addition, repetitive TMS (rTMS) modulates cortical excitability beyond the duration of the rTMS trains themselves. Depending on rTMS parameters, a lasting inhibition or facilitation of cortical excitability can be induced. These effects can be demonstrated neurophysiologically or by combining rTMS with neuroimaging techniques. The effects do not remain limited to the cortical area directly targeted by rTMS, but affect a wider neural network transynaptically. Modulation of cortical excitability by rTMS may in the future be useful not only as a research tool but also as a therapeutic intervention in neurology, psychiatry, and neurorehabilitation. PMID- 9736468 TI - Visual hemifield mapping using transcranial magnetic stimulation coregistered with cortical surfaces derived from magnetic resonance images. AB - The perception of a visual stimulus can be inhibited by occipital transcranial magnetic stimulation. This visual suppression effect has been attributed to disruption in the cortical gray matter of primary visual cortex or in the fiber tracts leading to V1 from the thalamus. However, others have suggested that the visual suppression effect is caused by disruption in secondary visual cortex. Here the authors used a figure-eight coil, which produces a focal magnetic field, and a Quadropulse stimulator to produce visual suppression contralateral to the stimulated hemisphere in five normal volunteer subjects. The authors coregistered the stimulation sites with magnetic resonance images in these same subjects using optical digitization. The stimulation sites were mapped onto the surface of the occipital lobes in three-dimensional reconstructions of the cortical surface to show the distribution of the visual suppression effect. The results were consistent with disruption of secondary visual cortical areas. PMID- 9736469 TI - Magnetic stimulation of visual cortex: factors influencing the perception of phosphenes. AB - Using transcranial magnetic stimulation of occipital cortex, the authors studied the stimulus parameters that generate phosphenes in healthy volunteers. Single pulses or trains of stimuli readily elicited phosphenes in all subjects. The threshold current needed to elicit perception of phosphenes was essentially the same for stimulus trains from 250 msec to 2000 msec in length, but increased dramatically for trains of shorter duration. The effect of stimulus frequency was variable, with each subject having a distinctive "frequency tuning curve," but overall, the threshold current necessary to produce phosphenes decreased as frequency of stimulation increased. Using paired pulses, the perceptual threshold was flat for interstimulus intervals between 2 msec and 100 msec, but increased rapidly as the interstimulus interval was increased above 100 msec. Stimulation of sites lateral to the midline elicited phosphenes in the contralateral visual field. Phosphenes were dominant in the lower and peripheral aspects of the visual fields. The findings are discussed in relation to similar studies of electrical stimulation of somatosensory cortex. PMID- 9736470 TI - Transcranial magnetic stimulation of the oculomotor and abducens nerves: determining the site of excitation in the cat. AB - The authors have attempted to stimulate the feline oculomotor and abducens nerves using a magnetic coil (MC) and to determine the optimal reliable MC position for eliciting motor evoked potentials. The authors have also determined the site of excitation to analyze the validity and potential advantages of this technique as a means of neurophysiologically studying the oculomotor and abducens nerves. The potentials of both of these muscles are evoked by MC placed to be symmetrical to the line connecting the vertex and the center of the eyeball on the side being examined, as the coil center corresponds to the midpoint of this line. The latencies of the motor responses of the superior rectus and lateral rectus were 1.08 +/- 0.22 and 1.02 +/- 0.21 msec, respectively, suggesting that magnetic stimulation excites both the oculomotor and the abducens nerve at the superior orbital fissure. PMID- 9736471 TI - Assessment of functional cerebral laterality for language using magnetoencephalography. AB - The purpose of the study was to examine the feasibility of using magnetoencephalography (MEG), a noninvasive functional brain imaging technique, to assess cerebral laterality for language. The magnetic flux normal to the scalp surface was measured with a whole-head neuromagnetometer while subjects (n = 16) were engaged in a word-matching and a tone-matching task. The effect of hemisphere and task on the number of satisfactory equivalent current dipole (ECD) solutions obtained during the late portion of the responses to the word and tone stimuli was examined. An interhemispheric ECD laterality index was also computed. Satisfactory ECD solutions were localized in perisylvian cortices during both tasks. A greater number of ECDs was found in the left hemisphere in 14 (87%) of 16 of the subjects in the word-matching task, a proportion that approaches the reported incidence of left-hemisphere dominance among right-handers. A similar proportion of subjects also showed a clear asymmetry in the number of ECDs favoring the left hemisphere in the language task as compared to the nonlanguage task. These findings suggest that MEG is a promising tool for laterality assessment. Magnetoencephalography-based functional asymmetry data are currently being compared against invasive presurgical procedures (i.e., intracarotid amobarbital procedure). PMID- 9736472 TI - Heart failure: a medical hydra. PMID- 9736473 TI - Epidemiology of heart failure and ventricular dysfunction. PMID- 9736474 TI - Why does the myocardium fail? Insights from basic science. PMID- 9736475 TI - Organisation of the care of patients with heart failure. PMID- 9736476 TI - Successes and failures of current treatment of heart failure. PMID- 9736477 TI - 15 years of heart-failure trials: what have we learned? PMID- 9736478 TI - New targets for heart-failure therapy: endothelin, inflammatory cytokines, and oxidative stress. PMID- 9736479 TI - Is preventive medicine responsible for the increasing prevalence of heart failure? PMID- 9736480 TI - Mechanism of induction of rat hepatic CYP2B and 3A by the pesticide methoxychlor. AB - Our earlier investigation demonstrated that methoxychlor treatment induced hepatic cytochrome P450 2B1/2B2 and 3A proteins in rats and enhanced their respective enzymatic activities. To determine the mechanism of the methoxychlor mediated induction and whether methoxychlor acts as a frank inducer and not merely by causing stabilization of the cytochrome P450 proteins or of their mRNAs, we assessed the effect of methoxychlor treatment on the transcriptional rate and mRNA levels of these hemeproteins by nuclear run-on and northern blot analyses. Methoxychlor treatment markedly elevated the transcriptional rates of cytochrome P4502B1/2B2 and 3A. Also, higher levels of mRNA of these CYPs were observed. By contrast, mRNA levels of CYP1A1/1A2 were not affected. Using a nuclear run-on method that apparently measures specifically the transcription initiation rate of CYP2B1 and 2B2, we observed that the transcriptional rate of 2B2 was increased substantially more than that of CYP2B1. This differential induction of CYP2B2 versus 2B1 may account for the discrepancy between the levels of induced CYP2B protein and the much lower-than-expected CYP2B-mediated enzymatic activity observed in our earlier study. PMID- 9736481 TI - Developmentally specific effects of the DNA cross-linking agent mitomycin C on phosphoenolpyruvate carboxykinase gene expression in vivo: correlation with changes in chromatin structure within the promoter region of the gene. AB - Previous studies in our laboratory have demonstrated that genotoxic chemical carcinogens have strong preferential effects on expression of certain inducible genes at nonovertly toxic doses in vivo. The effects of the DNA cross-linking agent and chemotherapy drug, mitomycin C (MMC), on expression of the developmental and hormone-regulated gene, phosphoenolpyruvate carboxykinase (PEPCK), were examined in chick embryo liver in vivo as a function of development and were compared with changes in the chromatin structure of the PEPCK gene promoter. The liver PEPCK gene was fully hormone inducible as early as 8 days of embryonic development but was refractory to MMC until after day 10. This onset of responsiveness to MMC was correlated with qualitative changes in the pattern of DNase I hypersensitive sites (DHS) within the PEPCK promoter. There was also a gradual decrease and then a complete loss of both hormone inducibility and MMC responsiveness between 14 and 17 days of development that was correlated with a quantitative change in the overall DNase sensitivity of the liver PEPCK gene promoter over this period. These results suggest that carcinogen sensitivity of the PEPCK gene is related to its ability to respond to its normal induction signals and that chromatin structure may play a central role in these effects. PMID- 9736482 TI - Activation of hepatic NF-kappaB by the herbicide Dicamba (2-methoxy-3,6 dichlorobenzoic acid) in female and male rats. AB - Nuclear factor-kappaB is a transcription factor that is activated in many different cell types by pathologic stimuli, such as reactive oxygen intermediates. One class of hepatocarcinogens, the peroxisome proliferators, may produce reactive oxygen intermediates, and one potent peroxisome proliferator, ciprofibrate, was recently reported to activate nuclear factor-kappaB. In this study, we investigated whether Dicamba, a broad leaf herbicide and peroxisome proliferator, could activate nuclear factor-KB in the livers of rats. Female and male Sprague Dawley rats (n = 4) were fed diets containing either 0, 1, or 3% Dicamba or 0.01% ciprofibrate for 7 days. As expected, the potent peroxisome proliferator, ciprofibrate, significantly increased fatty acyl CoA oxidase, peroxisomal beta-oxidation, and catalase activities in male rats and, except for catalase, also in female rats. Dicamba significantly increased peroxisomal fatty acyl CoA oxidase, peroxisomal beta-oxidation, and catalase activities, but decreased the activity of the cytosolic antioxidant enzyme, Se-dependent glutathione peroxidase, in both female and male rats. Dicamba increased nuclear factor-kappaB binding in the nuclear protein of livers from male rats fed both the 1 and 3% Dicamba diets. However, the highest binding was seen in nuclear protein from female rats fed 3% Dicamba. Both supershift and cold competition assays confirmed that this DNA binding activity was specific for nuclear factor kappaB. Our results in this study suggest that the herbicide and peroxisome proliferator Dicamba has the ability to activate nuclear factor-kappaB. PMID- 9736483 TI - Role of selenium against lead toxicity in male rats. AB - Male albino rats were intramuscularly administered a single dose of lead acetate (100 micromol/kg b.wt). Another group of rats were injected with sodium selenite (10 micromol/kg b.wt) before lead intoxication. After 3 and 24 hours, lead treatment resulted in significant increases in acid and alkaline phosphatases, GOT and GPT, total proteins, and cholesterol in serum. The total triglycerides in serum was decreased after 24 hours of intoxication. Lead treatment also produced significant elevation of lipid peroxidation in liver and kidney. The antioxidant capacity of hepatic and renal cells in terms of the activities of superoxide dismutase, glutathione reductase, and glutathione content was diminished. It appears from these results that lead may exert its toxic effect via peroxidative damage to renal and hepatic cell membranes after 24 hours. Selenium administration prior to lead injection produced pronounced prophylactic action against lead effects, and it is observed that selenium enhances the endogenous antioxidant capacity of the cells by increasing the activities of the superoxide dismutase and glutathione reductase and the glutathione content. As a result, the lipid peroxidation was decreased in both liver and kidney. PMID- 9736484 TI - Dichloroacetic acid induction of peroxisome proliferation in cultured hepatocytes. AB - Trichloroethylene is a widespread industrial solvent and one of the most common environmental contaminants. Trichloroethylene causes hepatocarcinoma in the B6C3F1 mouse in a dose-dependent manner. Trichloroethylene's hepatocarcinogenicity is thought to be mediated through its metabolites trichloroacetate and dichloroacetate. Although the mechanism of action is not well understood, hepatic tumors are thought to arise as a result of excessive peroxisome-dependent active oxygen production or secondary to enhanced cell replication. The peroxisome proliferative activity of trichloroacetate has been replicated in cultured rodent hepatocytes, while that of dichloroacetate has not been demonstrated. The present experiments were designed to characterize the peroxisome proliferative response to dichloroacetate in hepatocyte cultures from male B6C3F1 mice and male Long Evans rats. The cultured hepatocytes were treated after attachment with 0.1, 0.5, 1.0, 2.0, or 4.0 mM dichloroacetate for 72 hours. Peroxisome proliferation was assessed by measuring palmitoyl-CoA oxidation and by immunoquantitation of peroxisomal bifunctional enzyme. Palmitoyl CoA oxidation increased in a concentration-dependent manner, with maximal induction of 5.5- and 5-fold in mouse and rat hepatocytes, respectively, after treatment with 2.0 mM dichloroacetate. Peroxisomal bifunctional enzyme protein levels also increased in a concentration-dependent manner in both rat and mouse hepatocytes in response to dichloroacetate exposure. These results indicate that the peroxisomal response observed in vivo in response to dichloroacetate administration can be reproduced in primary cultures of rat and mouse hepatocytes treated with dichloroacetate. Further studies using this model system will help elucidate mechanisms of dichloroacetate-induced hepatocarcinogenesis. PMID- 9736485 TI - In vitro oxidation of the hydrolysis product of sulfur mustard, 2,2'-thiobis ethanol, by mammalian alcohol dehydrogenase. AB - The mechanism of vesication from sulfur mustard remains unknown in spite of 80 years of investigation. We recently reported sulfur mustard-related inhibition of one or more protein (serine/threonine) phosphatases in tissue cytosol in vitro, suggesting a mechanism common to other vesicants such as cantharidin and Lewisite. Our investigation showed that this inhibition was related to the concentration of 2,2'-thiobis-ethanol (thiodiglycol), the hydrolysis product of sulfur mustard, rather than to the concentration of mustard itself. Related work showed an increase in the rate of NAD (but not NADP) reduction upon the addition of thiodiglycol to mouse liver cytosol. This result provided evidence that metabolism beyond thiodiglycol may be contributing to protein phosphatase inhibition. This observation indicated that metabolism involving one or more dehydrogenases may be necessary to produce the ultimate inhibitor of the protein phosphatases. We report here that thiodiglycol is a substrate for horse liver alcohol dehydrogenase (Km = 3.68+/-0.45 mM and Vmax = 0.22 +/-0.01 micromol min( 1) mg protein(-1)) and for pyridine nucleotide-linked enzymes in mouse liver and human skin cytosol. The alcohol dehydrogenase-specific inhibitor 4-methylpyrazole inhibited the oxidation of thiodiglycol by the pure horse liver enzyme as well as by the enzymes in human skin and mouse liver cytosol, indicating that the activity in the tissue preparations is also alcohol dehydrogenase. PMID- 9736486 TI - Sequence-specific DNA minor groove binders. Design and synthesis of netropsin and distamycin analogues. PMID- 9736487 TI - Synthesis and in vitro T-cell immunogenicity of conjugates with dual specificity: attachment of epitope peptides of 16 and 38 kDa proteins from Mycobacterium tuberculosis to branched polypeptide. AB - T-cell epitope containing peptides covalently attached to macromolecular carriers can be considered as synthetic immunogens for the development of skin-test diagnostics and of vaccines. As a carrier, an amphoteric branched chain polypeptide, poly[Lys-(Glui-DL-Alam)] (EAK) with poly(L-lysine) backbone has been used. This polypeptide with free alpha-amino and gamma-carboxyl groups at the end of the side chains was conjugated with peptides representing two immunodominant regions of the 16 and 38 kDa proteins of Mycobacterium tuberculosis, respectively. Peptide C91SEFAYGSFVRTVSLPVGADE110 was elongated by Cys at the N terminal and attached to the carrier containing protected SH groups to form disulfide bridges. Peptide 65FNLWGPAFHERYPNVTITA83 was conjugated to the 3-(2 pyridyldithio)propionic acid N-hydroxysuccinimide ester (SPDP) modified and acetylated EAK by introducing amide bond between the free alpha-amino group of peptide and the free gamma-COOH group of Glu at the terminal position of the branches. This strategy lead to chemically well-defined synthetic immunogens that contain two different epitopes in multiple copies covalently linked to a synthetic branched polypeptide carrier. In vitro T-cell immunogenicity of a prototype conjugate was studied using T-cell hybridomas, lymph node cells from 38 kDa protein immunized mice, and human peripheral blood mononuclear cell (PBMC) cultures from sensitized individuals. These data document that the specific T cell stimulatory effect of each mycobacterial epitope was maintained in this conjugate. Taken together, these findings suggest that it is feasible to use a biodegradable polymeric polypeptide for producing macromolecular bioconjugates for the stimulation of T-cell responses. PMID- 9736488 TI - Catalytic properties of galactose oxidase to liposome-forming amphiphiles which have many pendent galactose residues. AB - A galactose-carrying vinyl monomer [2-(methacryloyloxy)ethyl beta-D galactopyranoside, MEGal] was polymerized by using a lipophilic radical initiator. The amphiphiles obtained (DODA-PMEGal) formed stable liposomes by mixing with phospholipids, and the galactose residues on the liposome surface were effectively recognized and oxidized by galactose oxidase. The affinity (estimated by the 1/Km value) of galactose oxidase for the galactose residues on the liposomes was higher than those for free galactose and MEGal and dependent on the length of galactose-carrying polymer chains on the liposome surface and the fluidity of the membranes, whereas not significantly influenced by the surface density of galactose residues on the liposomes. The affinity of galactose oxidase for the galactose-carrying linear polymers, which were prepared by using an ordinary azo-type radical initiator and a chain-transfer reagent, was also higher than those for free galactose and MEGal and dependent on the degree of polymerization of MEGal. The affinity was, however, relatively much smaller than those for DODA-PMEGals incorporated in liposomes. PMID- 9736489 TI - Sequence-specific photomodification of DNA by an oligonucleotide phenanthrodihydrodioxin conjugate. AB - We introduce a new member of a family of photochemically active oligonucleotide conjugates. A Phenanthrodihydrodioxin (PDHD)-based agent was synthesized and covalently linked to a 5'-end of the 9-mer oligonucleotide via a hexamethylene linker. The conjugate hybridized to a complementary 30-nucleotide-long target and efficiently cleaved it in a sequence specific manner. Up to 67% of target was specifically damaged (51% cross-links and 16% direct cleavage). While the photosensitizer alone nonspecifically damaged only Gs in a single-stranded target, its conjugate cross-linked to and damaged also A, T, and C sites in a target in agreement with duplex and triplex formation. PMID- 9736490 TI - Polycaprolactone-b-poly(ethylene oxide) block copolymer micelles as a novel drug delivery vehicle for neurotrophic agents FK506 and L-685,818. AB - Micelles formed from polycaprolactone-b-poly(ethylene oxide) (PCL-b-PEO) diblock copolymers were investigated as a novel drug delivery system. The affinity of the micelles for hydrophobic solubilizates was assayed by determining the partition coefficient for the lipophilic compound, pyrene, between the micelles and water; the partition coefficient was found to be on the order of 10(2). The Trypan blue and Alamar blue survival assays were used to assess the in vitro biocompatibility of the micelles with PC 12 cells, MCF-7 breast cancer cells, and primary cultures of human microglia, astrocytes, and cortical neurons. The micelles were then studied as a delivery vehicle for the neurotrophic agents FK506 and L-685,818 in PC 12 cell cultures. In both cases, the micelle-incorporated drugs, in the presence of nerve growth factor (5 ng/mL), were able to promote the degree of differentiation of the PC 12 rat pheochromocytoma cells. PMID- 9736491 TI - Liposome-anchored vascular endothelial growth factor aptamers. AB - Nuclease-resistant aptamers identified from randomized nucleic acid libraries represent a novel class of drug candidates. Aptamers are synthesized chemically and therefore can be readily modified with functional groups that modulate their properties. We report here on the preparation, initial characterization, and functional properties of a nuclease-resistant vascular endothelial growth factor (VEGF) aptamer anchored in liposome bilayers through a lipid group on the aptamer. While the high-affinity binding to VEGF is maintained, the plasma residence time of the liposome-anchored aptamer is considerably improved compared with that of the free aptamer. The lipid group attachment and/or liposome anchoring leads to a dramatic improvement in inhibitory activity of the aptamer toward VEGF-induced endothelial cell proliferation in vitro and vascular permeability increase and angiogenesis in vivo. PMID- 9736492 TI - A novel ternary ligand system for 99mTc-labeling of hydrazino nicotinamide modified biologically active molecules using imine-N-containing heterocycles as coligands. AB - A hydrazinonicotinamide-functionalized cyclic platelet glycoprotein IIb/IIIa (GPIIb/IIIa) receptor antagonist [HYNICtide, cyclo(D-Val-NMeArg-Gly-Asp-Mamb(5-(6 (6-hydrazinonicotina mido)hexanamide)))] was labeled with 99mTc using tricine and a series of imine-N-containing heterocycles as coligands. The imine-N-containing heterocycles include N-omega-Acetylhistamine (HIS-AC), N-(2 hydroxyethyl)isonicotinamide (ISONIC-HE), isonicotinic acid (ISONIC), isonicotinoyl-L-aspartic acid dimethyl ester (ISONIC-L-Asp-OMe2), 4-methyl-5 thiazoleethanol (MTE), nicotinic acid (NIC), 3-nitro-1,2,4-triazole (NTZ), 4 pyridylacetic acid (PA), 4-pyridineethanesulfonic acid (PES), and 3 pyridinesulfonic acid (PSA). The synthesis of these new ternary ligand [99mTc]HYNICtide complexes can be performed in one or two steps in high yield and high specific activity (>/=10 000 Ci/mmol HYNICtide). For example, the reaction of HYNICtide, [99mTc]pertechnetate, nicotinic acid, stannous chloride, and tricine at pH approximately 5 and 100 degreesC for 20 min results in the complex [99mTc(HYNICtide)(tricine)(NIC)] in >/=90% yield as determined by radio-HPLC. It was found that ternary ligand technetium complexes, [99mTc(HYNICtide)(tricine)(L)] (L = ISONIC, ISONIC-L-Asp-OMe2, ISONIC-HE, MTE, PA, PES, and PSA) are formed as equal mixtures of two isomeric forms. Complex [99mTc(HYNICtide)(tricine)(L)] (L = HIS-AC and NTZ) showed more than two well resolved radiometric peaks at the retention times of interest, suggesting that they may have more than two forms in solution due to different bonding modalities of HIS-AC and NTZ. By a chirality experiment, it was found that the presence of two radiometric peaks is a result of the resolution of the two diastereomers which are formed by the combination of the chiral HYNICtide and the chiral technetium chelate. The formation of two diastereomers was also observed when a chiral imine-N-containing coligand was used for the radiolabeling of HYNIC-BA. The new ternary ligand [99mTc]HYNICtide complexes were found to be stable for up to 6 h in the reaction mixture. The high solution stability is attributed to their kinetic inertness. The composition of these complexes was determined to be 1:1:1:1 for Tc:HYNICtide:L:tricine (L = imine-N-containing heterocycles) through a series of mixed ligand experiments on the tracer (99mTc) level. The lipophilicity of the ternary ligand [99mTc]HYNICtide complexes can be systematically varied by the choice of polyaminocarboxylate and imine-N containing coligands. Using the combination of tricine and an imine-N-containing coligand, HYNIC-derivatized peptides or other small molecules can be labeled with 99mTc in high specific activity and high stability for potential use as radiopharmaceuticals. PMID- 9736493 TI - Optimization and immunological characterization of a photochemically coupled lysergic acid diethylamide (LSD) immunogen. AB - A photoreactive heterobifunctional linker was used to prepare an immunogen in which lysergic acid diethylamide was indirectly coupled to keyhole limpet hemocyanin at multiple sites on the drug. It was possible to attach approximately 35 drug molecules to each protein using approximately equal amounts of both species during the reaction. The presence of buffer components or water severely compromised reaction efficiency, as estimated from the molar substitution ratio. Factors such as excess linker, pH, irradiation of dry matrix in the absence of buffer, and the drug/protein ratio used during photolysis were shown to have pronounced effects on reaction efficiency. Structural insights regarding immunogen coupling were obtained by determining the specificities of antibodies which were raised against the immunogen. Cross-reactivity data indicated that haptenation of protein likely occurred at positions N1 and N6 of lysergic acid diethylamide, which is plausible given the electrophilicity of the photogenerated aryl nitrene. PMID- 9736494 TI - Preparation of a spermine conjugate of the bis-phenanthroline ligand Clip-Phen and evaluation of the corresponding copper complex. AB - We described the synthesis of conjugates of N, N-diethylethylenediamine and spermine with Clip-Phen, a new ligand with two 1,10-phenanthrolines linked by serinol. The ability of the copper complexes of these Clip-Phen conjugates to cleave PhiX174 DNA when activated by a reductant in the presence of air was compared to that of 1,10-phenanthroline copper complexes. The results indicated that Clip-Phen-spermine with copper was the most active DNA cleaver. Comparison with a conjugate having only one phenanthroline unit linked to spermine showed that the high activity of Clip-Phen-spermine-Cu was not only due to the presence of spermine, which increases the affinity of the Clip-Phen entity for the DNA, but was effectively due to the presence of the two bridged phenanthroline entities. The DNA cleavage activity of (Clip-Phen-spermine)CuCl2 was analyzed on a 167 bp restriction fragment and a 45-mer double-stranded oligonucleotide. PMID- 9736495 TI - A polyethylene glycol copolymer for carrying and releasing multiple copies of cysteine-containing peptides. AB - Two different methods were developed to prepare an adduct of a poly(ethylene glycol)-lysine copolymer with either cysteamine or 1-amino-2-methyl-2 propanethiol. Cysteine-containing peptides could then be disulfide-linked to the thiol groups on the polymer in a facile manner. In the described procedures, a coupling ratio of about 8 peptides/molecule of poly(ethylene glycol)-lysine copolymer (Mw = 27 000) was typically attained. The products were stable at neutral pH, but the peptides could be released from the polymer in a physiologically relevant reducing environment. The release rate was highly dependent on the linker used for forming the disulfide bond. To illustrate the potential biomedical usefulness of this polymer carrier, a Tat peptide-PEG conjugate was shown to inhibit expression of a reporter gene fused to the TAR element of human immunodeficiency virus in a model cell assay. PMID- 9736496 TI - Cathepsin substrates as cleavable peptide linkers in bioconjugates, selected from a fluorescence quench combinatorial library. AB - Several extended peptide substrates for the human liver enzymes cathepsin B and cathepsin D have been selected as cleavable linkers for lysosomal proteolysis of bioconjugates. A one-bead-one-peptide combinatorial library of 9(4) fluorogenic substrates was employed. We designed this library to explore a set of substrates containing nonionizable/nonoxidizable groups to meet the requirements of prelabeling [Li et al. (1994) Bioconjugate Chem. 5, 101-104] as well as to yield stable conjugates whose preparation is straightforward. PMID- 9736498 TI - Closing Remarks. PMID- 9736497 TI - Modification of the thiourea linkage of a fluorescein-oligonucleotide conjugate to a guanidinium motif during ammonia deprotection. AB - During the preparation of a fluorescent conjugate based on a manganese cationic porphyrin carboxylate derivative linked to a 5'-amino-3'-fluorescein-labeled oligonucleotide, we observed the chemical transformation of the thiourea linkage to a guanidinium function during the deprotection in ammonia of the 3' fluorescein-oligonucleotide from CPG support. We also noted a secondary reactivity of the activated carboxylate group of the metalloporphyrin precursor with the fluorescein entity of the conjugated oligonucleotide. PMID- 9736500 TI - Master of Science in Epidemiology. PMID- 9736499 TI - Recipient of the Stokes Award in Preventive Cardiology. PMID- 9736527 TI - Nucleoside analog 1592U89 and human immunodeficiency virus protease inhibitor 141W94 are synergistic in vitro. AB - The use of combinations of anti-human immunodeficiency virus (anti-HIV) agents targeted to different molecular targets will most likely result in increased viral suppression and may also delay or prevent the emergence of resistant HIV strains. The purpose of the present study was to develop information on the in vitro anti-HIV activities of combinations of the reverse transcriptase inhibitor 1592U89 and the protease inhibitor 141W94 to help guide the choice of dosages in clinical trials. Triplicate in vitro dose-response matrices were prepared with MT 2 cells infected with HIV type 1 (HIV-1) strain IIIB. In order to account for the effects of protein binding, tissue culture medium with 10% fetal bovine serum was supplemented with the human serum proteins alpha1 acid glycoprotein (1 mg/ml) and albumin (40 mg/ml). The three-dimensional drug interaction surface for 1592U89 and 141W94 was constructed with the program MacSynergy II. As analyzed relative to a Bliss Independence null reference model, this combination was synergistic, with volumes of synergy exceeding 100 (99% confidence). Analysis of the data set with a fully parametric form of an equation for the quantitation of drug interaction developed by Greco et al. (W. R. Greco, G. Bravo, and J. C. Parsons, Pharmacol. Rev. 47:331-385, 1995) resulted in an interaction term statistically significantly greater than 0.0, indicating true synergy. Both methods concur that this combination is significantly synergistic. These data, with favorable findings from phase I/II trials for each drug alone, suggest that the combination of 1592U89 plus 141W94 should be further evaluated in clinical trials. PMID- 9736528 TI - Inhibitory action of a truncated derivative of the amphibian skin peptide dermaseptin s3 on Saccharomyces cerevisiae. AB - The inhibitory activity of a truncated derivative of the natural amphibian skin peptide dermaseptin s3-(1-16)-NH2 [DS s3 (1-16)] against Saccharomyces cerevisiae was studied. Significant growth inhibition was observed after exposure to 3.45 microgram of the peptide per ml at pH 6.0 and 7.0, with complete growth inhibition occurring at 8.63 microgram of peptide per ml for all pH values tested. Using confocal scanning laser microscopy, we have shown that DS s3 (1-16) disrupted the yeast cell membrane resulting in the gross permeabilization of the cell to the nuclear stain ethidium bromide. However, the principal inhibitory action of the peptide was not due to disruption of intracellular pH homeostasis. Instead, growth inhibition by the peptide correlated with the efflux of important cellular constituents such as ADP, ATP, RNA, and DNA into the surrounding medium. The combination of DS s3 (1-16) with mild heating temperatures as low as 35 degreesC significantly enhanced the inhibitory effect of the peptide (8.63 microgram/ml), and at 45 degreesC greater than 99% of the population was killed in 10 min. In summary, a derivative of a natural antimicrobial peptide has potential, either alone or in combination with mild heating, to prevent the growth of or kill spoilage yeast. PMID- 9736529 TI - Fungal beta-tubulin, expressed as a fusion protein, binds benzimidazole and phenylcarbamate fungicides. AB - Benzimidazoles are important antitubulin agents used in veterinary medicine and plant disease control. Resistance is a practical problem correlated with single amino acid changes in beta-tubulin and is often linked to greater sensitivity to phenylcarbamates. This negative cross-resistance creates opportunities for durable antiresistance strategies. Attempts to understand the molecular basis of benzimidazole resistance have been hampered by the inability to purify tubulin from filamentous fungi. We have overcome some of these problems by expressing beta-tubulin as a fusion with a maltose binding protein. This fusion protein is soluble, and we confirm for the first time using a gel filtration assay that benzimidazoles indeed bind to beta-tubulin. This binding is reduced by the mutation Glu198-->Gly198, which also confers resistance. Binding of phenylcarbamates is the complete opposite, reflecting their biological activity and the negative cross-resistance. This suggests that the fungicide binding sites fold correctly in the fusion protein. PMID- 9736530 TI - Pharmacokinetics and distribution over the blood-brain barrier of zalcitabine (2',3'-dideoxycytidine) and BEA005 (2', 3'-dideoxy-3'-hydroxymethylcytidine) in rats, studied by microdialysis. AB - Microdialysis was applied to sample the unbound drug concentration in the extracellular fluid in brain and muscle of rats given zalcitabine (2',3' dideoxycytidine; n = 4) or BEA005 (2', 3'-dideoxy-3'-hydroxymethylcytidine; n = 4) (50 mg/kg of body weight given subcutaneously). Zalcitabine and BEA005 were analyzed by high-pressure liquid chromatography with UV detection. The maximum concentration of zalcitabine in the dialysate (Cmax) was 31.4 +/- 5. 1 microM (mean +/- standard error of the mean) for the brain and 238. 3 +/- 48.1 microM for muscle. The time to Cmax was found to be from 30 to 45 min for the brain and from 15 to 30 min for muscle. Zalcitabine was eliminated from the brain and muscle with half-lives 1.28 +/- 0.64 and 0.85 +/- 0.13 h, respectively. The ratio of the area under the concentration-time curve (AUC) (from 0 to 180 min) for the brain and the AUC for muscle (AUC ratio) was 0.191 +/- 0.037. The concentrations of BEA005 attained in the brain and muscle were lower than those of zalcitabine, with Cmaxs of 5.7 +/- 1.4 microM in the brain and 61.3 +/- 12.0 microM in the muscle. The peak concentration in the brain was attained 50 to 70 min after injection, and that in muscle was achieved 30 to 50 min after injection. The half lives of BEA005 in the brain and muscle were 5.51 +/- 1.45 and 0.64 +/- 0.06 h, respectively. The AUC ratio (from 0 to 180 min) between brain and muscle was 0.162 +/- 0.026. The log octanol/water partition coefficients were found to be 1.19 +/- 0.04 and -1.47 +/- 0.01 for zalcitabine and BEA005, respectively. The degrees of plasma protein binding of zalcitabine (11% +/- 4%) and BEA005 (18% +/- 2%) were measured by microdialysis in vitro. The differences between zalcitabine and BEA005 with respect to the AUC ratio (P = 0.481), half-life in muscle (P = 0.279), and level of protein binding (P = 0.174) were not statistically significant. The differences were statistically significant in the case of the half-life in the brain (P = 0.032), clearance (P = 0.046), volume of distribution (P = 0.027) in muscle, and octanol/water partition coefficient (P = 0.019). PMID- 9736531 TI - Granulocyte colony-stimulating factor enhances endotoxin-induced decrease in biliary excretion of the antibiotic cefoperazone in rats. AB - We have recently reported that endotoxin (lipopolysaccharide [LPS]) derived from Klebsiella pneumoniae dramatically decreased the biliary excretion of the beta lactam antibiotic cefoperazone (CPZ), which is primarily excreted into the bile via the anion transport system, in rats. The present study was designed to investigate the effect of human recombinant granulocyte colony-stimulating factor (G-CSF), which is reported to be beneficial in experimental models of inflammation, on the pharmacokinetics and biliary excretion of CPZ in rats. CPZ (20 mg/kg of body weight) was administered intravenously 2 h after the intravenous injection of LPS (250 microgram/kg). G-CSF was injected subcutaneously at 12 microgram/kg for 3 days and was administered intravenously at a final dose of 50 microgram/kg 1 h before LPS injection. Peripheral blood cell numbers were also measured. LPS dramatically decreased the systemic and biliary clearances of CPZ and the bile flow rate. Pretreatment with G-CSF enhanced these decreases induced by LPS. The total leukocyte numbers were increased in rats pretreated with G-CSF compared to the numbers in the controls, while the total leukocyte numbers were decreased (about 3,000 cells/microliter) by treatment with LPS. Pretreatment with G-CSF produces a deleterious effect against the LPS-induced decrease in biliary secretion of CPZ, and leukocytes play an important role in that mechanism. PMID- 9736532 TI - Inhibitor-resistant OXY-2-derived beta-lactamase produced by Klebsiella oxytoca. AB - Klebsiella oxytoca strains are generally moderately resistant to amoxicillin and ticarcillin due to the activities of the chromosomally encoded OXY-1 and OXY-2 class A beta-lactamase families. These enzymes have the ability to hydrolyze not only penicillins but also cephalosporins, including cefuroxime, ceftriaxone, and aztreonam, and are inhibited by clavulanic acid. A Klebsiella oxytoca strain was isolated from a culture of blood from a patient who had been treated with amoxicillin-clavulanate (3 g/day) for 10 days 1 month earlier. This strain harbored an unusual phenotype characterized by resistance to amoxicillin clavulanate. It produced an OXY-2-type beta-lactamase (pI 6.3), as confirmed by PCR amplification with primers specific for the OXY-2-encoding gene. Gene sequencing revealed a point mutation (A-->G) corresponding to the amino acid substitution Ser-->Gly at position 130. This mutant enzyme was poorly inhibited by inhibitors, and its kinetic constants compared to those of the parent enzyme were characterized by an increased Km value for ticarcillin, with a drastically reduced activity against cephalosporins, as is observed with inhibitor-resistant TEM enzymes. The substitution Ser-->Gly-130 was previously described in the inhibitor-resistant beta-lactamase SHV-10 derived from an SHV-5 variant, but this is the first report of such a mutant in OXY enzymes from K. oxytoca. PMID- 9736533 TI - Pharmacodynamic analysis of the activity of quinupristin-dalfopristin against vancomycin-resistant Enterococcus faecium with differing MBCs via time-kill-curve and postantibiotic effect methods. AB - Quinupristin-dalfopristin (Q-D) is a new water-soluble, semisynthetic antibiotic that is derived from natural streptogramins and that is combined in a 30:70 ratio. A number of studies have described the pharmacodynamic properties of this drug, but most have investigated only staphylococci or streptococci. We evaluated the relationship between Q-D, quinupristin (Q), and/or dalfopristin (D) susceptibility parameters and antibacterial activities against 22 clinical isolates of vancomycin-resistant Enterococcus faecium (VREF) by using the concentration-time-kill-curve method and by measuring postantibiotic effects. Q D, Q, and D MICs and minimum bactericidal concentrations (MBCs) ranged from 0.125 to 1 and 0.25 to 64, 8 to 512 and >512, and 2 to 8 and 8 to 512 microgram/ml, respectively. There were no significant relationships between susceptibilities to the individual components and the susceptibilities to the Q-D combination product. In the time-kill-curves studies, Q-D at a concentration of 6 microgram/ml was at least bacteriostatic against all VREF tested. There was increased activity against more susceptible isolates when the isolates were grouped either by Q-D MBCs or by Q MICs. By multivariate regression analyses, the percent change in the inoculum from that at the baseline was significantly correlated with the Q MIC (R = 0.74; P = 0.008) and the Q-D concentration-to-MBC ratio (R = 0.58; P = 0.02) and was inversely correlated with the Q-D MBC-to-MIC ratio (R = 0.68; P = 0.003). A strong correlation existed between the killing rate and the Q-D concentration-to-MBC ratio (R = 0.99; P < 0.0001). Time to 99.9% killing was best correlated with the Q-D MBC (R = 0.96; P < 0.0001). The postantibiotic effect ranged from 0.2 to 3.2 h and was highly correlated with the Q-D concentration-to-MBC ratio (R = 0.96; P < 0.0001) and was less highly correlated with the Q MIC (R = 0.42; P = 0.04). Further study of these relationships with in vitro or in vivo infection models that simulate Q-D pharmacokinetics should further define the utility of these pharmacodynamic parameters in the prediction of Q-D activity for the treatment of VREF infections in humans. PMID- 9736534 TI - Sparfloxacin resistance in clinical isolates of Streptococcus pneumoniae: involvement of multiple mutations in gyrA and parC genes. AB - Antimicrobial susceptibility testing revealed among 150 clinical isolates of Streptococcus pneumoniae 4 pneumococcal isolates with resistance to fluoroquinolones (MIC of ciprofloxacin, >/=32 microgram/ml; MIC of sparfloxacin, >/=16 microgram/ml). Gene amplification and sequencing analysis of gyrA and parC revealed nucleotide changes leading to amino acid substitutions in both GyrA and ParC of all four fluoroquinolone-resistant isolates. In the case of strains 182 and 674 for which sparfloxacin MICs were 16 and 64 microgram/ml, respectively, nucleotide changes were detected at codon 81 in gyrA and codon 79 in parC; these changes led to an Ser-->Phe substitution in GyrA and an Ser-->Phe substitution in ParC. Strains 354 and 252, for which sparfloxacin MICs were 128 microgram/ml, revealed multiple mutations in both gyrA and parC. These strains exhibited nucleotide changes at codon 85 leading to a Glu-->Lys substitution in GyrA, in addition to Ser-79-->Tyr and Lys-137-->Asn substitutions in ParC. Moreover, strain 252 showed additional nucleotide changes at codon 93, which led to a Trp- >Arg substitution in GyrA. These results suggest that sparfloxacin resistance could be due to the multiple mutations in GyrA and ParC. However, it is possible that other yet unidentified mutations may also be involved in the high-level resistance to fluoroquinolones in S. pneumoniae. PMID- 9736535 TI - Molecular mode of action of the antifungal beta-amino acid BAY 10-8888. AB - BAY 10-8888 is a cyclic beta-amino acid that is related to cispentacin and that has antifungal activity. Candida albicans cells accumulated BAY 10-8888 intracellularly to a concentration about 200 that in the medium when grown in media with a variety of nitrogen sources. In complex growth medium, BAY 10-8888 transport activity was markedly reduced and was paralleled by a decrease in its antifungal activity. Uptake of BAY 10-8888 was mediated by an H+-coupled amino acid transporter with specificity for branched-chain amino acids (isoleucine, leucine, and valine) and showed a KT (Michaelis constant of the transport reaction) of 0.95 mM and a Vmax of 18.9 nmol x min-1 x 10(7) cells-1. Similar to the transport of natural amino acids in Saccharomyces cerevisiae, the transport of BAY 10-8888 into the cell was unidirectional. Efflux occurred by diffusion and was not carrier mediated. Inside the cell BAY 10-8888 inhibited specifically isoleucyl-tRNA synthetase, resulting in inhibition of protein synthesis and cell growth. Intracellular isoleucine reversed BAY 10-8888-induced growth inhibition. BAY 10-8888 was not incorporated into proteins. BAY 10-8888 inhibited isoleucyl tRNA synthetase with the same concentration dependency as protein biosynthesis in intact cells assuming 200-fold accumulation. PMID- 9736536 TI - Activities of LL-37, a cathelin-associated antimicrobial peptide of human neutrophils. AB - Human neutrophils contain two structurally distinct types of antimicrobial peptides, beta-sheet defensins (HNP-1 to HNP-4) and the alpha-helical peptide LL 37. We used radial diffusion assays and an improved National Committee for Clinical Laboratory Standards-type broth microdilution assay to compare the antimicrobial properties of LL-37, HNP-1, and protegrin (PG-1). Although generally less potent than PG-1, LL-37 showed considerable activity (MIC, <10 microgram/ml) against Pseudomonas aeruginosa, Salmonella typhimurium, Escherichia coli, Listeria monocytogenes, Staphylococcus epidermidis, Staphylococcus aureus, and vancomycin-resistant enterococci, even in media that contained 100 mM NaCl. Certain organisms (methicillin-resistant S. aureus, Proteus mirabilis, and Candida albicans) were resistant to LL-37 in media that contained 100 mM NaCl but were susceptible in low-salt media. Burkholderia cepacia was resistant to LL-37, PG-1, and HNP-1 in low- or high-salt media. LL-37 caused outer and inner membrane permeabilization of E. coli ML-35p. Chromogenic Limulus assays revealed that LL 37 bound to E. coli O111:B4 lipopolysaccharide (LPS) with a high affinity and that this binding showed positive cooperativity (Hill coefficient = 2.02). Circular dichroism spectrometry disclosed that LL-37 underwent conformational change in the presence of lipid A, transitioning from a random coil to an alpha helical structure. The broad-spectrum antimicrobial properties of LL-37, its presence in neutrophils, and its inducibility in keratinocytes all suggest that this peptide and its precursor (hCAP-18) may protect skin and other tissues from bacterial intrusions and LPS-induced toxicity. The potent activity of LL-37 against P. aeruginosa, including mucoid and antibiotic-resistant strains, suggests that it or related molecules might have utility as topical bronchopulmonary microbicides in cystic fibrosis. PMID- 9736538 TI - New trimethoprim-resistant dihydrofolate reductase cassette, dfrXV, inserted in a class 1 integron. AB - The nucleotide sequence of a plasmid-borne trimethoprim resistance gene from a commensal fecal Escherichia coli isolate revealed a new dihydrofolate reductase gene, dfrXV, which occurred as a gene cassette integrated in a site-specific manner in a class 1 integron. The new gene shows 84% nucleotide identity and the predicted protein shows 90% amino acid identity with dfrI and DHFR type I, respectively. Genes for spectinomycin resistance, aadA1 [ant (3")-Ia], and sulfonamide resistance, sulI, were located downstream of dfrXV in a manner identical to that in pLMO229. PMID- 9736537 TI - Glycopeptide antibiotic resistance genes in glycopeptide-producing organisms. AB - The mechanism of high-level resistance to vancomycin in enterococci consists of the synthesis of peptidoglycan terminating in D-alanyl-D-lactate instead of the usual D-alanyl-D-alanine. This alternate cell wall biosynthesis pathway is ensured by the collective actions of three enzymes: VanH, VanA, and VanX. The origin of this resistance mechanism is unknown. We have cloned three genes encoding homologs of VanH, VanA, and VanX from two organisms which produce glycopeptide antibiotics: the A47934 producer Streptomyces toyocaensis NRRL 15009 and the vancomycin producer Amycolatopsis orientalis C329.2. The predicted amino acid sequences are highly similar to those found in VRE: 54 to 61% identity for VanH, 59 to 63% identity for VanA, and 61 to 64% identity for VanX. Furthermore, the orientations of the genes, vanH, vanA, and vanX, are identical to the orientations found in vancomycin-resistant enterococci. Southern analysis of total DNA from other glycopeptide-producing organisms, A. orientalis 18098 (chloro-eremomycin producer), A. orientalis subsp. lurida (ristocetin producer), and Amycolatopsis coloradensis subsp. labeda (teicoplanin and avoparcin producer), with a probe derived from the vanH, vanA, and vanX cluster from A. orientalis C329.2 revealed cross-hybridizing DNA in all strains. In addition, the vanH, vanA, vanX cluster was amplified from all glycopeptide-producing organisms by PCR with degenerate primers complementary to conserved regions in VanH and VanX. Thus, this gene sequence is common to all glycopeptide producers tested. These results suggest that glycopeptide-producing organisms may have been the source of resistance genes in vancomycin-resistant enterococci. PMID- 9736539 TI - Influence of the TonB energy-coupling protein on efflux-mediated multidrug resistance in Pseudomonas aeruginosa. AB - TonB couples the energized state of the cytoplasmic membrane to the operation of outer membrane receptors responsible for Fe(III) siderophore uptake across the outer membrane of gram-negative bacteria. A tonB mutant of Pseudomonas aeruginosa deficient in iron siderophore uptake was shown in the present study to be hypersusceptible to a wide variety of antibiotics, reminiscent of the phenotype of mutants defective in the mexAB-oprM antibiotic efflux operon. This was not related to influences of a tonB mutation on the iron status of the cell, and indeed, intrinsic antibiotic susceptibility and mexAB-oprM expression were unaffected by iron levels in the growth medium. The presence of tonB on a multicopy plasmid increased the level of resistance of a MexAB-OprM+ strain but not that of a MexAB-OprM- strain to a variety of antimicrobial agents. mexAB-oprM expression was not, however, altered in a tonB deletion mutant, indicating that any influence of TonB on MexAB-OprM-mediated multidrug resistance was at the level of pump activity. Consistent with this, drug accumulation assays revealed that the tonB deletion mutant exhibited decreased levels of drug efflux. Still, the multidrug resistance of a nalB strain was not wholly abrogated by a tonB mutation, indicating that it is likely not an essential component of the efflux apparatus. Similarly, elimination of tonB from an nfxB strain only partially compromised MexCD-OprJ-mediated multidrug resistance. Intriguingly, the drug susceptibility of a mexAB-oprM deletion strain was increased following deletion of tonB, suggesting that TonB may also influence antibiotic resistance mediated by determinants other than MexAB-OprM (and MexCD-OprJ). Thus, TonB plays an important role in both intrinsic and acquired antibiotic resistance in P. aeruginosa. PMID- 9736540 TI - In vitro effectiveness of povidone-iodine on Acanthamoeba isolates from human cornea. AB - Acanthamoeba keratitis is a severe ocular infection secondary to accidental macro or microscopic trauma of the cornea. Starting in 1985, a dramatic increase of this infection was recorded along with the spread of contact lens use. This protozoal disease is difficult to treat because of the scarcity of efficacious topical and systemic drugs. We evaluated the in vitro effectiveness of povidone iodine (PVP-I [Betadine]), an agent with broad antibacterial and antiviral activity, compared to that of chlorhexidine (CXD), a cationic antiseptic, on Acanthamoeba isolates from patients with amebic keratitis. The results showed that PVP-I solution from 0.5 to 2.5% has a better antiamebic activity both on trophic and cystic stages of Acanthamoeba spp. than does CXD. PMID- 9736541 TI - Pharmacokinetic profiles of high-dose intravenous ciprofloxacin in severe sepsis. The Baragwanath Ciprofloxacin Study Group. AB - The pharmacokinetics of 400 mg of ciprofloxacin given intravenously (i.v.) every 8 h (q8h) in severely septic adults was documented in a multidisciplinary, tertiary referral intensive care unit (ICU). Sixteen evaluable patients (three pharmacokinetic profiles) without renal dysfunction and with severe sepsis were studied. Ciprofloxacin at a dosage of 400 mg given i.v. q8h was administered over 1 h. Plasma samples for assay (high-pressure liquid chromatography) were taken at timed intervals (preinfusion, at the end of infusion, and at 1, 2, 3, 5, and 7 h postinfusion) for first-dose kinetics (day 0 [D0]), D2, and between D6 and D8. All pharmacokinetic variables were calculated by noncompartmental methods. Standard intensive care was provided. Peak ciprofloxacin concentrations were as follows: D0, 6. 01 +/- 1.93 mg/liter; D2, 6.68 +/- 2.01 mg/liter; and D6 to D8 6.45 +/- 1.54 mg/liter. Trough levels were as follows: D0, 0.6 +/- 0.5 mg/liter; D2, 0.7 +/- 0.4 mg/liter; and D6 to D8 0.6 +/- 0.4 mg/liter. The areas under the concentration curves (8 h) were as follows: D0, 13.3 +/- 3.8 mg . h/liter; D2, 16.8 +/- 5.4 mg . h/liter; and D6 to D8, 15.5 +/- 4.7 mg . h/liter. No drug related serious adverse events occurred. For 17 of 18 patients enrolled in the study, the causative organisms were susceptible to ciprofloxacin. One patient developed renal failure (non-drug related) after the administration of three doses of ciprofloxacin. One patient was infected with ciprofloxacin-resistant organisms on enrollment. Nine of 16 evaluable patients had clinical cures, and 8 had bacteriological cures. One patient developed a ciprofloxacin-resistant superinfection. In two patients the clinical course was indeterminate. Two bacteriological failures occurred. We conclude that in critically ill adults ciprofloxacin at a dosage of 400 mg given i.v. q8h is safe. Its pharmacokinetic profile provides bactericidal activity against most organisms encountered in an ICU. Except for some initial accumulation on D2, no further accumulation occurred in patients without renal failure. Ciprofloxacin should be administered i.v. at a dosage of 400 mg q8h for severe sepsis. PMID- 9736542 TI - Incidence of foscarnet resistance and cidofovir resistance in patients treated for cytomegalovirus retinitis. The Cytomegalovirus Retinitis and Viral Resistance Study Group. AB - Cytomegalovirus (CMV) retinitis is a common opportunistic infection in patients with AIDS. With long-term therapy for CMV retinitis, resistant CMV may develop. In a prospective study of 122 patients with CMV retinitis, 2.4 and 0.8% of patients had foscarnet-resistant blood culture isolates (50% inhibitory concentration [IC50], >400 microM) and urine culture isolates, respectively, at diagnosis of CMV retinitis prior to treatment, whereas 4.1 and 6.6% had cidofovir resistant (IC50, >2 microM) blood and urine culture isolates, respectively. Patients were treated according to best medical judgement. Of 44 foscarnet treated patients, 26% had a resistant blood or urine culture isolate by 6 months of treatment and 37% had a resistant isolate by 9 months; of 13 cidofovir-treated patients, 29% had a resistant blood or urine culture isolate by 3 months of therapy. The probabilities of developing foscarnet resistance while on foscarnet and developing cidofovir resistance while on cidofovir were not significantly different from that for developing ganciclovir resistance while on ganciclovir (odds ratios, 1.87 [P = 0.19] and 2.28 [P = 0.15], respectively). PMID- 9736543 TI - Human immunodeficiency virus type 1 cDNA integration: new aromatic hydroxylated inhibitors and studies of the inhibition mechanism. AB - Integration of the human immunodeficiency virus type 1 (HIV-1) cDNA is a required step for viral replication. Integrase, the virus-encoded enzyme important for integration, has not yet been exploited as a target for clinically useful inhibitors. Here we report on the identification of new polyhydroxylated aromatic inhibitors of integrase including ellagic acid, purpurogallin, 4,8, 12 trioxatricornan, and hypericin, the last of which is known to inhibit viral replication. These compounds and others were characterized in assays with subviral preintegration complexes (PICs) isolated from HIV-1-infected cells. Hypericin was found to inhibit PIC assays, while the other compounds tested were inactive. Counterscreening of these and other integrase inhibitors against additional DNA-modifying enzymes revealed that none of the polyhydroxylated aromatic compounds are active against enzymes that do not require metals (methylases, a pox virus topoisomerase). However, all were cross-reactive with metal-requiring enzymes (restriction enzymes, a reverse transcriptase), implicating metal atoms in the inhibitory mechanism. In mechanistic studies, we localized binding of some inhibitors to the catalytic domain of integrase by assaying competition of binding by labeled nucleotides. These findings help elucidate the mechanism of action of the polyhydroxylated aromatic inhibitors and provide practical guidance for further inhibitor development. PMID- 9736544 TI - Antimalarial synergy of cysteine and aspartic protease inhibitors. AB - It has been proposed that the Plasmodium falciparum cysteine protease falcipain and aspartic proteases plasmepsin I and plasmepsin II act cooperatively to hydrolyze hemoglobin as a source of amino acids for erythrocytic parasites. Inhibitors of each of these proteases have potent antimalarial effects. We have now evaluated the antimalarial effects of combinations of cysteine and aspartic protease inhibitors. When incubated with cultured P. falciparum parasites, cysteine and aspartic protease inhibitors exhibited synergistic effects in blocking parasite metabolism and development. The inhibitors also demonstrated apparent synergistic inhibition of plasmodial hemoglobin degradation both in culture and in a murine malaria model. When evaluated for the treatment of murine malaria, a combination of cysteine and aspartic protease inhibitors was much more effective than higher concentrations of either compound used alone. These results support a model whereby plasmodial cysteine and aspartic proteases participate in the degradation of hemoglobin, and they suggest that combination antimalarial therapy with inhibitors of the two classes of proteases is worthy of further study. PMID- 9736545 TI - Influence of assay methodology on the measurement of free serum ceftriaxone concentrations. AB - The influence of assay methodology on the measurement of the active free fraction of ceftriaxone in plasma was determined. The free fraction was measured by three methods: agar diffusion bioassay, precipitation of plasma protein with methanol followed by high-performance liquid chromatography (HPLC) of the supernatant, and ultrafiltration of plasma followed by HPLC of the filtrate. In human serum, the free ceftriaxone levels were significantly lower (P = 0.03) when measured on ultrafiltrates compared to the other two methods. This difference disappeared when dolphin serum was studied. After ultrafiltration, human serum was shown, by Scatchard plot analysis, to have two ceftriaxone binding sites. Species differences were also demonstrated. Hence, in humans, determination of free plasma ceftriaxone varies with the assay method employed. PMID- 9736546 TI - Treatment of community-acquired acute uncomplicated urinary tract infection with sparfloxacin versus ofloxacin. The Sparfloxacin Multi Center UUTI Study Group. AB - The efficacy and safety of a 3-day regimen of sparfloxacin were compared with those of a 3-day regimen of ofloxacin for the treatment of community-acquired acute uncomplicated urinary tract infections. Four hundred nineteen women were enrolled in a randomized, open-label, observer-blinded, multicenter study; 204 received sparfloxacin as a 400-mg loading dose on the first day and 200 mg once daily thereafter, and 215 received ofloxacin as 200 mg twice daily. A total of 383 patients met the criteria for clinical evaluability, and 174 were also bacteriologically evaluable; all treated patients were included in the safety analysis. Escherichia coli (86%) and Staphylococcus saprophyticus (4.6%) were the organisms most commonly isolated. Positive clinical responses were obtained 5 to 9 days after therapy in more than 92% of the patients in each group; sustained clinical cure rates 4 to 6 weeks after therapy were 78.3 and 76.9% in the sparfloxacin and ofloxacin groups, respectively. A positive bacteriologic response was observed in 98% of the bacteriologically evaluable patients in each treatment group at 5 to 9 days posttherapy and in 88.2 and 92.6% of the patients in the sparfloxacin and ofloxacin groups, respectively, 4 to 6 weeks after therapy. Almost 90% of all adverse events were of mild or moderate severity; the most frequent events at least possibly related to drug treatment were those common to the fluoroquinolones, namely, nausea, diarrhea, headache, insomnia, and photosensitivity. Photosensitivity was more frequent in the sparfloxacin group (6.9% versus 0.5% in the ofloxacin group); insomnia was more frequent in the ofloxacin group (3.7% versus 1.0% in the sparfloxacin group). These data suggest that a once-daily, 3-day regimen of sparfloxacin is effective and generally well tolerated in the treatment of acute uncomplicated urinary tract infections. PMID- 9736548 TI - Sordarins: A new class of antifungals with selective inhibition of the protein synthesis elongation cycle in yeasts. AB - GR135402, a sordarin derivative, was isolated in an antifungal screening program. GR135402, sordarin, and derivatives of both compounds were evaluated for their ability to inhibit cell-free translational systems from five different pathogenic fungi (Candida albicans, Candida glabrata, Candida krusei, Candida parapsilosis, and Cryptococcus neoformans). The activity profile of GR135402 is extended to other chemical compounds derived from sordarin. Experimental results indicate that sordarin analogs exert their antifungal effects by specifically inhibiting the protein synthesis elongation cycle in yeasts but do not affect protein synthesis machinery in mammalian systems. Intrinsically resistant strains owe their resistance to differences in the molecular target of sordarins in these strains. Preliminary studies performed to elucidate the mode of action of this new class of antifungal agents have shown that the putative target of sordarins is one of the protein synthesis elongation factors. PMID- 9736547 TI - Association of a thr-371 substitution in a conserved amino acid motif of penicillin-binding protein 1A with penicillin resistance of Streptococcus pneumoniae. AB - We determined the nucleotide sequence between 1,903 and 3,097 bp of pbp1a, which encodes the transpeptidase domain of PBP 1A, from clinical isolates of penicillin resistant Streptococcus pneumoniae (PRSP) serotypes 19 (n = 8), 6 (n = 9), 23 (n = 6), and 14 (n = 2) and two penicillin-susceptible S. pneumoniae (PSSP) isolates. These serotyped PRSP strains were isolated predominantly in Japan from 1993 through 1997. The 25 resistant strains were classified into five groups on the basis of the extent of sequence differences. Strains in groups I (n = 5; serotype 6), II (n = 3; serotype 19), and III (n = 12; different serotypes) had sequences highly homologous to the sequence of pbp1a of PRSP strains from South Africa, Spain, and the United States. The group IV strain (n = 1; serotype 14) had unique deletions from or insertions in the sequences. The sequences of group V strains (n = 4; serotypes 6 and 23) had relatively few differences from the sequences of the PSSP strains. For strains (n = 18) for which the threonine at codon 371 (Thr-371) in a conserved STMK motif of PBP 1A was substituted with an alanine or a serine (in addition to having altered pbp2x and pbp2b genes), penicillin MICs were >/= 1.0 microgram/ml. The PBPs 1A of these strains showed decreased affinities for [3H]benzylpenicillin and slightly faster mobilities on sodium dodecyl sulfate-polyacrylamide gels. In contrast, for strains (n = 4) without a substitution at Thr-371 in PBP 1A but with mutations in both pbp2x and pbp2b, penicillin MICs were 0.125 to 0.25 microgram/ml, and the affinities of their PBPs 1A were similar to that of PSSP PBPs 1A. Furthermore, for the Thr-371 substituted strains (n = 3) with altered pbp2x genes but native pbp2b genes, penicillin MICs were 0.125 to 0.25 microgram/ml. These results suggest that amino acid substitution of Thr-371 contributes to the development of penicillin resistance in PRSP strains with altered pbp2x and pbp2b genes. PMID- 9736549 TI - Identification of elongation factor 2 as the essential protein targeted by sordarins in Candida albicans. AB - The target for sordarins in Candida albicans has been elucidated. Kinetic experiments of sordarin inhibition as well as displacement experiments showed that the formation of a sordarin-target complex follows a reversible mechanism. Binding of tritiated drug to the target is enhanced in the presence of ribosomes. Isolation of the target by classical protein purification methods has allowed us to identify it as elongation factor 2. This is in agreement with the nature of sordarin derivatives as specific inhibitors of the elongation cycle within protein synthesis in yeasts. PMID- 9736550 TI - Two 2-hydroxy-3-alkyl-1,4-naphthoquinones with in vitro and in vivo activities against Toxoplasma gondii. AB - Two 3-alkyl-substituted 2-hydroxy-1,4-naphthoquinones, NSC 113452 (NSC52) and NSC 113455 (NSC55), were evaluated for activity against Toxoplasma gondii in vitro and in murine models of acute toxoplasmosis. In vitro, both NSC52 and NSC55 significantly inhibited intracellular replication of T. gondii. In vivo, each compound was examined alone and in combination with other drugs currently used for treatment of human toxoplasmosis. Although none of the compounds protected mice against death when administered orally, both were significantly protective when administered intraperitoneally. In addition, a significant increase in survival was observed when suboptimal doses of each compound were used in combination with suboptimal doses of pyrimethamine or sulfadiazine. These results indicate that combinations of NSC52 or NSC55 with pyrimethamine or sulfadiazine have promising activity against T. gondii. PMID- 9736551 TI - In vitro inactivation of Chlamydia trachomatis by fatty acids and monoglycerides. AB - The antichlamydial effects of several fatty acids and monoglycerides were studied by incubating Chlamydia trachomatis bacteria with equal volumes of lipid solutions for 10 min and measuring the reduction in infectivity titer compared with that in a control solution without lipid. Caprylic acid (8:0), monocaprylin (8:0), monolaurin (12:0), myristic acid (14:0), palmitoleic acid (16:1), monopalmitolein (16:1), oleic acid (18:1), and monoolein (18:1) at concentrations of 20 mM (final concentration, 10 mM) had negligible effects on C. trachomatis. In contrast, lauric acid (12:0), capric acid (10:0), and monocaprin (10:0) caused a greater than 10,000-fold (>4-log10) reduction in the infectivity titer. When the fatty acids and monoglycerides were further compared at lower concentrations and with shorter exposure times, lauric acid was more active than capric acid and monocaprin was the most active, causing a greater than 100, 000-fold (>5-log10) inactivation of C. trachomatis at a concentration of 5 mM for 5 min. The high levels of activity of capric and lauric acids and particularly that of monocaprin are notable and suggest that these lipids have specific antichlamydial effects. The mode of action of monocaprin was further studied by removal of the lipid by centrifugation before inoculation of Chlamydia onto host cells and by electron microscopy. The results indicate that the bacteria are killed by the lipid, possibly by disrupting the membrane(s) of the elementary bodies. A 50% effective concentration of 30 microgram/ml was found by incubation of Chlamydia with monocaprin for 2 h. The rapid inactivation of large numbers of C. trachomatis organisms by monocaprin suggests that it may be useful as a microbicidal agent for the prevention of the sexual transmission of C. trachomatis. PMID- 9736552 TI - Effect of food on the bioavailability of stavudine in subjects with human immunodeficiency virus infection. AB - A randomized, three-way crossover study was carried out to determine the effects of food ingestion on the pharmacokinetics of stavudine (d4T). Fifteen subjects with human immunodeficiency virus (HIV) infection and CD4(+) cell counts of >/=200/microliter received 70 mg of d4T in a fasting state or 1 h before or 5 min after a standardized high-fat breakfast. A 7- to 15-day washout period was included between treatments. Blood and urine were collected before and for 10 h after dosing, and plasma and urine d4T concentrations were determined with a validated radioimmunoassay. Plasma drug concentration-time data were analyzed with a noncompartmental model. The mean maximum plasma drug concentration (Cmax) and the time to Cmax (Tmax) for administration of d4T after a meal were significantly lower and longer (P = 0.0001 for both measures) than those observed in the fasting state, although the area under the concentration-time curve from time zero to infinity (AUC0-infinity) was not significantly different. Neither of these parameters was significantly altered when d4T was taken 1 h before a meal. The bioavailability of d4T taken after a meal was 95% of that observed in the fasting state, and it was 97% when d4T was administered before a meal (P > 0.05 for both comparisons with the fasting state). The results of this study indicate that (i) ingestion of food does not affect the bioavailability of d4T and that patients with HIV infection can take it without regard to meals, and (ii) absorption is essentially complete within 1 h when d4T is administered in the fasted state. PMID- 9736553 TI - Activity of voriconazole combined with neutrophils or monocytes against Aspergillus fumigatus: effects of granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor. AB - Voriconazole (VCZ) was tested for antifungal activity against Aspergillus fumigatus hyphae alone or in combination with neutrophils or monocytes. Antifungal activity was measured as percent inhibition of hyphal growth in assays using the dye MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] or XTT [2, 3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxa nilide ]. With both assays, VCZ inhibited hyphal growth at concentrations of <1 microgram/ml and was almost as active as amphotericin B. VCZ (0.6 microgram/ml) was sporicidal, as was amphotericin B (0.4 microgram/ml). With both the MTT and XTT assays, neutrophils alone inhibited hyphae; when combined with VCZ, there was additive activity. Both granulocyte colony-stimulating factor- and granulocyte macrophage colony-stimulating factor (GM-CSF)-treated polymorphonuclear neutrophils (PMN) had enhanced inhibition of hyphal growth. Moreover, such treatment of PMN also enhanced the collaboration of PMN with VCZ. Monocytes inhibited hyphal growth. When VCZ was combined with monocytes or monocytes were treated with GM-CSF, inhibition was significantly increased, to similar levels. However, the combination of VCZ with GM-CSF treatment of monocytes did not significantly increase the high-level inhibition by monocytes with either agent alone. PMID- 9736554 TI - Alterations in topoisomerase IV and DNA gyrase in quinolone-resistant mutants of Mycoplasma hominis obtained in vitro. AB - Mycoplasma hominis mutants were selected stepwise for resistance to ofloxacin and sparfloxacin, and their gyrA, gyrB, parC, and parE quinolone resistance determining regions were characterized. For ofloxacin, four rounds of selection yielded six first-, six second-, five third-, and two fourth-step mutants. The first-step mutants harbored a single Asp426-->Asn substitution in ParE. GyrA changes (Ser83-->Leu or Trp) were found only from the third round of selection. With sparfloxacin, three rounds of selection generated 4 first-, 7 second-, and 10 third-step mutants. In contrast to ofloxacin resistance, GyrA mutations (Ser83 ->Leu or Ser84-->Trp) were detected in the first-step mutants prior to ParC changes (Glu84-->Lys), which appeared only after the second round of selection. Further analysis of eight multistep-selected mutants of M. hominis that were previously described (2) revealed that they carried mutations in ParE (Asp426- >Asn), GyrA (Ser83-->Leu) and ParE (Asp426-->Asn), GyrA (Ser83-->Leu) and ParC (Ser80-->Ile), or ParC (Ser80-->Ile) alone, depending on the fluoroquinolone used for selection, i.e., ciprofloxacin, norfloxacin, ofloxacin, or pefloxacin, respectively. These data indicate that in M. hominis DNA gyrase is the primary target of sparfloxacin whereas topoisomerase IV is the primary target of pefloxacin, ofloxacin, and ciprofloxacin. PMID- 9736555 TI - Detection of Tn917-like sequences within a Tn916-like conjugative transposon (Tn3872) in erythromycin-resistant isolates of Streptococcus pneumoniae. AB - A series of macrolide-lincosamide-streptogramin B (MLS)-resistant pneumococcal isolates of a variety of serotypes was examined and was found to contain Tn917 like elements by DNA-DNA hybridization. Like Tn1545, Tn917 also encodes an ermAM gene but does not mediate resistance to other antimicrobial agents. Furthermore, nucleotide sequence analyses of the DNAs flanking three of the Tn917-like elements revealed that they were inserted into orf9 of a Tn916-like element in a composite transposon-like structure (Tn3872). Other MLS-resistant strains appeared to contain Tn1545-like elements that had suffered a deletion of sequences including the aphA-3 sequences responsible for kanamycin resistance. Thus, the MLS resistance phenotype in pneumococci appears to be mediated by the ermAM present on a much wider variety of genetic elements than was previously appreciated. PMID- 9736556 TI - Characterization of a PSE-4 mutant with different properties in relation to penicillanic acid sulfones: importance of residues 216 to 218 in class A beta lactamases. AB - Class A beta-lactamases are inactivated by the suicide inactivators sulbactam, clavulanic acid, and tazobactam. An examination of multiple alignments indicated that amino acids 216 to 218 differed among class A enzymes. By random replacement mutagenesis of codons 216 to 218 in PSE-4, a complete library consisting of 40,864 mutants was created. The library of mutants with mutations at positions 216 to 218 in PSE-4 was screened on carbenicillin and ampicillin with the inactivator sulbactam; a collection of 14 mutants was selected, and their bla genes were completely sequenced. Purified wild-type and mutant PSE-4 beta lactamases were used to measure kinetic parameters. One enzyme, V216S:T217A:G218R, was examined for its peculiar pattern of inhibition. There was an increase in the Km from 68 microM for the wild type to 271 microM for the mutant for carbenicillin and 33 to 216 microM for ampicillin. Relative to the wild-type PSE-4 enzyme, 37- and 30-fold increases in Ki values were observed for the mutant enzyme for sulbactam and tazobactam, respectively. The results that were obtained suggested that positions 216 to 218 are important for interactions with penicillanic acid sulfone inhibitors. In contrast, V216 and A217 in the TEM 1 class A beta-lactamase do not tolerate amino acid residue substitutions. However, for the PSE-4 beta-lactamase, 11 of 14 mutants from the library of mutants with mutations at positions 216 to 218 whose sequences were determined had substitutions at position 216 (G, R, A, S) and position 217 (A, S). The data showed the importance of residues 216 to 218 in their atomic interactions with inactivators in the PSE-4 beta-lactamase structure. PMID- 9736557 TI - Rapid ganciclovir susceptibility assay using flow cytometry for human cytomegalovirus clinical isolates. AB - Rapid, quantitative, and objective determination of the susceptibilities of human cytomegalovirus (HCMV) clinical isolates to ganciclovir has been assessed by an assay that uses a fluorochrome-labeled monoclonal antibody to an HCMV immediate early antigen and flow cytometry. Analysis of the ganciclovir susceptibilities of 25 phenotypically characterized clinical isolates by flow cytometry demonstrated that the 50% inhibitory concentrations (IC50s) of ganciclovir for 19 of the isolates were between 1.14 and 6.66 microM, with a mean of 4.32 microM (+/-1.93) (sensitive; IC50 less than 7 microM), the IC50s for 2 isolates were 8.48 and 9.79 microM (partially resistant), and the IC50s for 4 isolates were greater than 96 microM (resistant). Comparative analysis of the drug susceptibilities of these clinical isolates by the plaque reduction assay gave IC50s of less than 6 microM, with a mean of 2.88 microM (+/-1.40) for the 19 drug-sensitive isolates, IC50s of 6 to 8 microM for the partially resistant isolates, and IC50s of greater than 12 microM for the four resistant clinical isolates. Comparison of the IC50s for the drug-susceptible and partially resistant clinical isolates obtained by the flow cytometry assay with the IC50s obtained by the plaque reduction assay showed an acceptable correlation (r2 = 0.473; P = 0.001), suggesting that the flow cytometry assay could substitute for the more labor-intensive, subjective, and time-consuming plaque reduction assay. PMID- 9736558 TI - Effects of alpha-thalassemia on pharmacokinetics of the antimalarial agent artesunate. AB - Thalassemia is common in Southeast Asia, where artemisinin derivatives are frequently used in the treatment of malaria. It has been previously reported that artemisinin derivatives can be concentrated by uninfected thalassemic erythrocytes in vitro but not by normal erythrocytes. As a follow-up to this report, we studied the antimalarial kinetics of intravascular artesunate (2.4 mg/kg of body weight) in 10 persons with normal hemoglobins and in 10 patients with thalassemia (2 with alpha-thalassemia type 1-hemoglobin Constant Spring and 8 with alpha-thalassemia type 1-alpha-thalassemia type 2). Concentrations of artesunate and its active metabolites in plasma were measured by bioassay and expressed relative to those of dihydroartemisinin, the major biologically active metabolite. Concentrations of intravascular artesunate in plasma peaked in both the normal individuals and the thalassemic individuals 15 min after injection (the first time point). Plasma drug concentrations at all time intervals, except that at 1 h, were significantly higher in thalassemic subjects than in normal subjects (P < 0.05). The area under the concentration-time curve was 9-fold higher (P < 0.001) and the volume of distribution at steady state was 15-fold lower (P < 0.001) in thalassemic than in normal subjects. In light of the potential neurotoxicity of artemisinin derivatives, these results suggest that thalassemic subjects may need a drug administration regimen different from that of normal patients. PMID- 9736559 TI - Inhibition of tumor necrosis factor alpha alters resistance to Mycobacterium avium complex infection in mice. AB - Increased production of tumor necrosis factor alpha (TNF-alpha) appears to play an important role in the progression of human immunodeficiency virus disease. One treatment strategy being explored is the use of TNF-alpha inhibitors. TNF-alpha also appears to be important in conferring resistance to infections, and the inhibition of this cytokine may exacerbate the emergence of opportunistic pathogens, such as Mycobacterium avium complex (MAC). The present study examines the possibility that inhibition of TNF-alpha will increase the progression of disease in mice infected with MAC. C57BL/6 beige (bg/bg) mice have been shown to be highly susceptible to infection with MAC and are routinely used for testing of antimycobacterial drugs. However, bg/bg mice are known to exhibit impaired phagocyte and natural killer cell function. Since these cell types are important sources of TNF-alpha, the susceptibility of the bg/bg strain to infection with MAC was compared with those of the heterozygous (bg/+) and wild-type (+/+) strains of C57BL/6 mice. The susceptibilities of the bg/bg and bg/+ strains of mice infected with MAC were found to be comparable. The +/+ strain was the least susceptible. Mycobacterial burden and serum TNF-alpha levels increased over time in all the strains of mice tested. The bg/+ strain of C57BL/6 mice was then chosen to measure the activity of TNF-alpha antagonists. Treatment with dexamethasone decreased serum TNF-alpha levels and increased mycobacterial burden. Treatment with anti-TNF-alpha antibody or pentoxifylline did not significantly alter serum TNF-alpha levels but increased mycobacterial burden. Treatment with thalidomide neither consistently altered mycobacterial burden in the spleens or livers of infected mice nor affected serum TNF-alpha levels. PMID- 9736560 TI - Synergy of nitric oxide and azoles against Candida species in vitro. AB - The candidacidal activity of nitric oxide (NO) as delivered by a class of compounds termed diazeniumdiolates has been investigated. Diazeniumdiolates are stable agents capable of releasing NO in a biologically usable form at a predicted rate, and three such compounds were examined for activity. One compound, (Z)-1-[N-(2-aminoethyl)-N-(2-ammonioethyl)amino]diazen-1- ium-1, 2 diolate (DETA-NO), proved to be most suitable for examining NO activity due to its relatively long half-life (20 h) and because of limited candidacidal activity of the uncomplexed DETA nucleophile. DETA-NO was active against six species of Candida for which the MICs necessary to inhibit 50% growth (MIC50s) ranged from 0.25 to 1.0 mg/ml. C. parapsilosis and C. krusei were the most susceptible to the compound. In addition to a determination of NO effects alone, the complex was utilized to investigate the synergistic potential of released NO in combination with ketoconazole, fluconazole, and miconazole. Activity was investigated in vitro against representative strains of Candida albicans, C. krusei, C. parapsilosis, C. tropicalis, C. glabrata, and C. dubliniensis. Determination of MIC50, MIC80 and MICs indicated that DETA-NO inhibits all strains tested, with strains of C. parapsilosis and C. krusei being consistently the most sensitive. The combination of DETA-NO with each azole was synergistic against all strains tested as measured by fractional inhibitory concentration indices that ranged from 0.1222 to 0.4583. The data suggest that DETA-NO or compounds with similar properties may be useful in the development of new therapeutic strategies for treatment of Candida infections. PMID- 9736561 TI - In vitro activity of lumefantrine (benflumetol) against clinical isolates of Plasmodium falciparum in Yaounde, Cameroon. AB - The in vitro antimalarial activity of the new Chinese synthetic drug, lumefantrine, also known as benflumetol (a fluorene derivative belonging to the aminoalcohol class), was determined by an isotopic microtest against 61 fresh clinical isolates of Plasmodium falciparum and compared with that of other established antimalarial agents. The geometric mean 50% inhibitory concentration of lumefantrine was 11.9 nmol/liter (95% confidence intervals, 10.4 to 13.6 nmol/liter; range, 3.3 to 25.6 nmol/liter). The in vitro activities of lumefantrine against the chloroquine-sensitive and the chloroquine-resistant isolates did not differ (P > 0.05). There was a significant positive correlation of responses between lumefantrine and two other aminoalcohols studied, mefloquine (r = 0.688) and halofantrine (r = 0.677), and between lumefantrine and artesunate (r = 0.420), suggesting a potential for in vitro cross-resistance. Our data suggest high in vitro activity of lumefantrine, comparable to that of mefloquine, and are in agreement with the promising results of preliminary clinical trials. PMID- 9736562 TI - Salmonella enteritidis: AmpC plasmid-mediated inducible beta-lactamase (DHA-1) with an ampR gene from Morganella morganii. AB - DHA-1, a plasmid-mediated cephalosporinase from a single clinical Salmonella enteritidis isolate, conferred resistance to oxyimino-cephalosporins (cefotaxime and ceftazidime) and cephamycins (cefoxitin and moxalactam), and this resistance was transferable to Escherichia coli HB101. An antagonism was observed between cefoxitin and aztreonam by the diffusion method. Transformation of the transconjugant E. coli strain with plasmid pNH5 carrying the ampD gene (whose product decreases the level of expression of ampC) resulted in an eightfold decrease in the MIC of cefoxitin. A clone with the same AmpC susceptibility pattern with antagonism was obtained, clone E. coli JM101(pSAL2-ind), and its nucleotide sequence was determined. It contained an open reading frame with 98. 7% DNA sequence identity with the ampC gene of Morganella morganii. DNA sequence analysis also identified a gene upstream of ampC whose sequence was 97% identical to the partial sequence of the ampR gene (435 bp) from M. morganii. The gene encoded a protein with an amino-terminal DNA-binding domain typical of transcriptional activators of the LysR family. Moreover, the intercistronic region between the ampC and ampR genes was 98% identical to the corresponding region from M. morganii DNA. AmpR was shown to be functional by enzyme induction and a gel mobility-shift assay. An ampG gene was also detected in a Southern blot of DNA from the S. enteritidis isolate. These findings suggest that this inducible plasmid-mediated AmpC type beta-lactamase, DHA-1, probably originated from M. morganii. PMID- 9736563 TI - Absence of effect of rufloxacin on theophylline pharmacokinetics in steady state. AB - Several quinolone antibacterial agents are known to inhibit the metabolism of theophylline, with the potential to cause adverse events due to raised theophylline concentrations during coadministration. A randomized crossover study was therefore conducted with 12 healthy male volunteers (ages, 23 to 34 years; body weight, 64 to 101 kg) to evaluate a possible interaction between rufloxacin and theophylline. Both drugs were administered at steady state. Following the administration of an oral loading dose of 400 mg on day 1, rufloxacin was given orally at 200 mg once daily on days 2 to 7 during one period only. During both periods, 146 mg of theophylline was administered orally twice daily for 3 days (which were days 4 to 6 of the rufloxacin coadministration period) and intravenously once the next morning to test for an interaction. Theophylline and rufloxacin concentrations were measured by reversed-phase high-pressure liquid chromatography, the pharmacokinetics of theophylline at steady state following administration of the last dose were calculated by compartment-model-independent methods. To compare the treatments, analysis of variance-based point estimates and 90% confidence intervals (given in parentheses) were calculated for the mean ratios of the pharmacokinetic parameters from the test (rufloxacin coadministration) over those from the reference (theophylline without rufloxacin) period. These were as follows: maximum concentration at steady state, 1.01 (0.96 to 1.07); area under the concentration-time curve from 0 to 12 h, 0.98 (0.94 to 1.02); half-life, 0.99 (0.95 to 1.03); total clearance at steady state, 1. 02 (0.99 to 1.06); and volume of distribution in the elimination phase, 1.01 (0.97 to 1.05). In conclusion, rufloxacin did not affect theophylline pharmacokinetics at steady state. Therefore, therapeutic coadministration of rufloxacin and theophylline is not expected to cause an increased incidence of theophylline related adverse events. PMID- 9736564 TI - In vitro pharmacodynamic studies of L-749,345 in comparison with imipenem and ceftriaxone against gram-positive and gram-negative bacteria. AB - L-749,345 is a new parenteral carbapenem with a very long half-life similar to that of ceftriaxone. The aim of the present study was to investigate different pharmacodynamic parameters of L-749,345 in comparison with those of ceftriaxone and imipenem. The following studies were performed: (i) comparative studies of the MICs of L-749, 345, imipenem, and ceftriaxone for 70 strains of gram-positive and gram-negative bacteria; (ii) comparative studies of the rate of killing of gram-positive and gram-negative bacteria by L-749,345, imipenem, and ceftriaxone; (iii) studies of the postantibiotic effects of L-749,345, imipenem, and ceftriaxone; and (iv) studies of the postantibiotic sub-MIC effects of L-749,345, imipenem, and ceftriaxone. Significantly lower MICs of L-749,345 compared with those of ceftriaxone were found for all gram-negative organisms except Haemophilus influenzae. The MICs of L-749,345 were similar to those of imipenem for all organisms except Pseudomonas aeruginosa and methicillin-resistant Staphylococcus aureus, for which the MICs of L-749,345 were higher. A concentration-dependent killing of methicillin-resistant S. aureus but not methicillin-susceptible strains was noted for both L-749,345 and imipenem. All three of the investigated drugs exhibited a postantibiotic effect against the gram-positive strains but exhibited no postantibiotic effect against the gram negative strains. PMID- 9736565 TI - Efficacy of nikkomycin Z in the treatment of murine histoplasmosis. AB - Immune-competent ICR and BALB/c athymic (nude) mice were infected intravenously with Histoplasma capsulatum and treated with either fluconazole or nikkomycin Z or 5% dextrose (controls). In immune-competent ICR mice, fluconazole and nikkomycin Z both prolonged survival when given at 5 mg/kg of body weight twice daily. When administered in doses as low as 2.5 mg/kg twice daily, nikkomycin Z reduced fungal counts in both the spleen and liver. When both drugs were combined, there was no antagonism, and in combined therapy spleen and liver counts were reduced more than for either drug alone. However, nikkomycin Z had no effect on brain fungal burden. In nude mice fluconazole and nikkomycin Z had an additive effect in prolongation of survival and reduction of liver and spleen burden. Nikkomycin Z is well tolerated, is at least as effective as fluconazole, and may interact beneficially with fluconazole for treatment of murine histoplasmosis. PMID- 9736566 TI - In vivo activities of amoxicillin and amoxicillin-clavulanate against Streptococcus pneumoniae: application to breakpoint determinations. AB - The in vivo activities of amoxicillin and amoxicillin-clavulanate against 17 strains of Streptococcus pneumoniae with penicillin MICs of 0.12-8.0 mg/liter were assessed in a cyclophosphamide-induced neutropenic murine thigh infection model. Renal impairment was produced by administration of uranyl nitrate to prolong the amoxicillin half-life in the mice from 21 to 65 min, simulating human pharmacokinetics. Two hours after thigh infection with 10(5) to 10(6) CFU, groups of mice were treated with 7 mg of amoxicillin per kg of body weight alone or combined with clavulanate (ratio, 4:1) every 8 h for 1 and 4 days. There was an excellent correlation between the MIC of amoxicillin (0.03 to 5.6 mg/liter) and (i) the change in log10 CFU/thigh at 24 h and (ii) survival after 4 days of therapy. Organisms for which MICs were 2 mg/liter or less were killed at 1.4 to 4.2 and 1.6 to 4.1 log10 CFU/thigh at 24 h by amoxicillin and amoxicillin clavulanate, respectively. The four strains for which MICs were >4 mg/liter grew 0.2 to 2.6 and 0.6 to 2. 3 logs at 24 h despite therapy with amoxicillin and amoxicillin-clavulanate, respectively. Infection was uniformly fatal by 72 h in untreated mice. Amoxicillin therapy resulted in no mortality with organisms for which MICs were 1 mg/liter or less, 20 to 40% mortality with organisms for which MICs were 2 mg/liter, and 80 to 100% mortality with organisms for which MICs were 4.0-5.6 mg/liter. Lower and higher doses (0.5, 2, and 20 mg/kg) of amoxicillin were studied against organisms for which MICs were near the breakpoint. These studies demonstrate that a reduction of 1 log10 or greater in CFU/thigh at 24 h is consistently observed when amoxicillin levels exceed the MIC for 25 to 30% of the dosing interval. These studies would support amoxicillin (and amoxicillin clavulanate) MIC breakpoints of 1 mg/liter for susceptible, 2 mg/liter for intermediate, and 4 mg/liter for resistant strains of S. pneumoniae. PMID- 9736567 TI - Safety, pharmacokinetics, and antiretroviral activity of intravenous 9-[2-(R) (Phosphonomethoxy)propyl]adenine, a novel anti-human immunodeficiency virus (HIV) therapy, in HIV-infected adults. AB - 9-[2-(R)-(Phosphonomethoxy)propyl]adenine (PMPA) is a nucleotide analogue with potent antiretroviral activity in vitro and in simian models. A randomized, double-blind, placebo-controlled, dose-escalation clinical trial of intravenous PMPA monotherapy was conducted in 20 human immunodeficiency virus (HIV)-infected adults with CD4 cell counts of >/=200 cells/mm3 and plasma HIV RNA levels of >/=10,000 copies/ml. Two dose levels were evaluated (1 and 3 mg/kg of body weight/day). Ten subjects were enrolled at each dose level (eight randomized to receive PMPA and two randomized to receive placebo). On day 1, a single dose of PMPA or placebo was administered by intravenous infusion. Beginning on study day 8, PMPA or placebo was administered once daily for an additional 7 consecutive days. All subjects tolerated dosing without significant adverse events. Mean peak serum PMPA concentrations were 2.7 +/- 0.9 and 9.1 +/- 2.1 microgram/ml in the 1- and 3-mg/kg cohorts, respectively. Serum concentrations declined in a biexponential fashion, with a terminal half-life of 4 to 8 h. At 3 mg/kg/day, a single infusion of PMPA resulted in a 0.4 log10 median decline in plasma HIV RNA by study day 8. Following 7 consecutive days of study drug administration thereafter, the median changes in plasma HIV RNA from baseline were -1.1, -0.6, and 0.1 log10 in the 3-mg/kg/day, 1-mg/kg/day, and placebo dose groups, respectively. Following the final dose in the 3-mg/kg/day cohort, the reduction in HIV RNA was sustained for 7 days before returning toward baseline. Further studies evaluating an oral prodrug of PMPA are under way. PMID- 9736568 TI - Dynamics of clarithromycin and azithromycin efficacies against experimental Haemophilus influenzae pulmonary infection. AB - The dynamics of clarithromycin and azithromycin efficacy against pulmonary Haemophilus influenzae infection in rats were evaluated. Efficacy was measured by reduction in pulmonary H. influenzae burden on days 3 and 7 postinoculation. Clarithromycin therapy was effective on day 3 or 7 of therapy, while azithromycin was effective on day 7 but not on day 3 of therapy. Both macrolides produced marked efficacy against all six strains of H. influenzae tested, including four strains for which MICs were above the susceptible breakpoint (8 microgram/ml) concentration of clarithromycin. The two macrolides demonstrated markedly different pharmacokinetic characteristics, with clarithromycin present in both blood and tissue, while azithromycin was concentrated primarily in tissue. During pulmonary infection in rats, H. influenzae was found in both intracellular locations and an extracellular location in the lung. Blood concentrations of clarithromycin and azithromycin approximated human pharmacokinetics, and the blood concentrations for either macrolide rarely exceeded MICs for H. influenzae. At dosages producing blood concentrations similar to values achieved clinically, clarithromycin produced efficacy on day 3 of therapy, while both clarithromycin and azithromycin were equally effective on day 7. The different dynamics of clarithromycin and azithromycin suggest that length of therapy should be considered as a key parameter in evaluations of drug efficacy. PMID- 9736569 TI - Safety, tolerance, and pharmacokinetics of a small unilamellar liposomal formulation of amphotericin B (AmBisome) in neutropenic patients. AB - The safety, tolerance, and pharmacokinetics of a small unilamellar liposomal formulation of amphotericin B (AmBisome) administered for empirical antifungal therapy were evaluated for 36 persistently febrile neutropenic adults receiving cancer chemotherapy and bone marrow transplantation. The protocol was an open label, sequential-dose-escalation, multidose pharmacokinetic study which enrolled a total of 8 to 12 patients in each of the four dosage cohorts. Each cohort received daily doses of either 1.0, 2.5, 5.0, or 7.5 mg of amphotericin B in the form of AmBisome/kg of body weight. The study population consisted of patients between the ages of 13 and 80 years with neutropenia (absolute neutrophil count, <500/mm3) who were eligible to receive empirical antifungal therapy. Patients were monitored for safety and tolerance by frequent laboratory examinations and the monitoring of infusion-related reactions. Efficacy was assessed by monitoring for the development of invasive fungal infection. The pharmacokinetic parameters of AmBisome were measured as those of amphotericin B by high-performance liquid chromatography. Noncompartmental methods were used to calculate pharmacokinetic parameters. AmBisome administered as a 1-h infusion in this population was well tolerated and was seldom associated with infusion-related toxicity. Infusion related side effects occurred in 15 (5%) of all 331 infusions, and only two patients (5%) required premedication. Serum creatinine, potassium, and magnesium levels were not significantly changed from baseline in any of the dosage cohorts, and there was no net increase in serum transaminase levels. AmBisome followed a nonlinear dosage relationship that was consistent with reticuloendothelial uptake and redistribution. There were no breakthrough fungal infections during empirical therapy with AmBisome. AmBisome administered to febrile neutropenic patients in this study was well tolerated, was seldom associated with infusion-related toxicity, was characterized by nonlinear saturation kinetics, and was effective in preventing breakthrough fungal infections. PMID- 9736570 TI - Antifungal activity of the pradimicin derivative BMS 181184 in the treatment of experimental pulmonary aspergillosis in persistently neutropenic rabbits. AB - The activity of the pradimicin derivative BMS 181184 was evaluated in a model of invasive pulmonary aspergillosis in persistently neutropenic rabbits and compared with that of amphotericin B deoxycholate. BMS 181184 at total daily doses of 50 and 150 mg/kg of body weight was at least as effective as amphotericin B at 1 mg/kg once a day in conferring survival and had comparable activity in reducing organism-mediated tissue injury and excess lung weight. Although treatment at all dosing regimens of BMS 181184 resulted in significant reductions in fungal tissue burden compared to untreated controls, equivalence to amphotericin B occurred only at the higher dosage level. Similar observations were made in bronchoalveolar lavage fluid cultures obtained postmortem. Monitoring of the animals through ultrafast computerized tomography scan revealed a marked resolution of pulmonary lesions during treatment with BMS 181184. The compound was well tolerated at all dosing regimens, and no toxicity was noted. Pharmacokinetic studies revealed nonlinear drug disposition with increased clearance at higher dosages and some evidence for extravascular drug accumulation. BMS 181184 had excellent activity in the treatment of experimental invasive pulmonary aspergillosis in persistently neutropenic rabbits, thus underscoring the potential of pradimicin derivatives in therapy of invasive aspergillosis in the neutropenic host. PMID- 9736571 TI - Pharmacokinetics of pentoxifylline and its metabolites in healthy mice and in mice infected with Candida albicans. AB - Pentoxifylline has immunomodulatory properties and has been shown to decrease organ damage and improve survival in animals with gram-negative sepsis or endotoxemia. This effect is mediated by a reduction in endotoxin-induced production of tumor necrosis factor alpha (TNF-alpha) by the host. In earlier studies, we observed an unexpected increase in mortality in mice infected with Candida albicans that were given pentoxifylline even though concentrations of TNF alpha in serum were not affected. The current study was designed to determine whether the pharmacokinetics of pentoxifylline and its metabolites were altered in C. albicans-infected mice and, if so, whether these changes could have contributed to the increased mortality. Noninfected mice and mice infected with C. albicans were treated with pentoxifylline (60 mg/kg of body weight) intraperitoneally every 8 h. Serum was collected from animals after one (day 0), four (day 1), or seven (day 2) injections of pentoxifylline or saline (controls). The first dose was administered 6 h after C. albicans infection. Serum was pooled. Concentrations of pentoxifylline and metabolites I, IV, and V were determined by capillary gas chromatography. Renal function and hepatic profiles were assessed. Pharmacokinetic parameters (maximum concentration of pentoxifylline in serum, half-life, and area under the concentration-time curve from 0 h to infinity [AUC(0)-infinity]) for all noninfected mice were similar and did not differ from those for day 0-infected mice. For day 1-infected mice, values of these three pharmacokinetic parameters for pentoxifylline and metabolite I were increased two- to fourfold over values for noninfected and day 0-infected mice. For metabolites IV and V, the AUC(0)-infinity was increased approximately eightfold over control values. In addition, day 1-infected mice demonstrated evidence of renal and hepatic dysfunction. In summary, C. albicans infection produced marked changes in the pharmacokinetics of pentoxifylline and its metabolites in the mice. The high concentrations of pentoxifylline and its metabolites in serum attained in infected mice may have contributed to the increased mortality of mice with systemic candidiasis. PMID- 9736572 TI - Effect of disposition of mannich antimalarial agents on their pharmacology and toxicology. AB - The use of the antimalarial agent amodiaquine has been curtailed due to drug induced idiosyncratic reactions. These have been attributed to the formation of a protein-reactive quinoneimine species via oxidation of the 4-aminophenol group. Therefore, the effects of chemical modifications on the disposition of amodiaquine in relation to its metabolism, distribution, and pharmacological activity have been investigated. The inclusion of a group at the C-5' position of amodiaquine reduced or eliminated bioactivation, as determined by glutathione conjugate formation in vivo. This can be seen in two series of C-5'-substituted compounds: the bis-Mannich antimalarial agents, including cycloquine and pyronaridine, and mono-Mannich antimalarial agents containing a 5'-chlorophenyl group (tebuquine and 5'-ClPAQ). Chemical substitution at the C-5' position also resulted in compounds which underwent slower elimination (<5% of the dose excreted into bile and urine, compared with 50% for amodiaquine) and increased levels of accumulation in tissue (10% of the dose in the liver at 48 h compared with 1% with amodiaquine). This may be due to an increase in either the lipophilicity or the basicity of the analogs and may reflect the lack of metabolic clearance for these compounds. The alteration in the disposition following the introduction of the C-5' substituent resulted in an increased duration of antimalarial activity in the mouse compared with that for amodiaquine. While this is desirable in the treatment of malaria, repeated administration for prophylaxis may induce toxicity through accumulation. Therefore, by simple chemical modification it is possible to block the bioactivation of amodiaquine while maintaining and in some cases extending the duration of antimalarial activity. PMID- 9736573 TI - Single-dose pharmacokinetics of meropenem during continuous venovenous hemofiltration. AB - The pharmacokinetic properties of meropenem were investigated in nine critically ill patients treated by continuous venovenous hemofiltration (CVVH). All patients received one dose of 1 g of meropenem intravenously. High-flux polysulfone membranes were used as dialyzers. Meropenem levels were measured in plasma and ultrafiltrate by high-performance liquid chromatography. The total body clearance and elimination half-life were 143.7 +/- 18.6 ml/min and 2.46 +/- 0.41 h, respectively. The post- to prehemofiltration ratio of meropenem was 0.24 +/- 0.06. Peak plasma drug concentrations measured 60 min postinfusion were 28.1 +/- 2.7 microgram/ml, and trough levels after 6 h of CVVH were 6.6 +/- 1.5 microgram/ml. The calculated total daily meropenem requirement in these patients with acute renal failure and undergoing CVVH was 2,482 +/- 321 mg. Based on these data, we conclude that patients with severe infections who are undergoing CVVH can be treated effectively with 1 g of meropenem every 8 h. PMID- 9736575 TI - Prevalence and characterization of the mechanisms of macrolide, lincosamide, and streptogramin resistance in isolates of Streptococcus pneumoniae. AB - Of a total of 147 erythromycin-resistant Streptococcus pneumoniae isolates, 64 (43.5%) were resistant to erythromycin, clindamycin, and streptogramin B (MLSB phenotype), 57 of which possessed the ermB gene. Eighty-two (55.8%) were resistant to erythromycin alone (M phenotype), 81 of which possessed the mefE gene. One was erythromycin and streptogramin B resistant but susceptible to clindamycin (MS phenotype) and possessed neither the erm gene nor the mefE gene. PMID- 9736574 TI - Pharmacokinetics of meropenem in critically ill patients with acute renal failure treated by continuous hemodiafiltration. AB - The pharmacokinetics of meropenem were studied in nine anuric critically ill patients treated by continuous venovenous hemodiafiltration. Peak levels after infusion of 1,000 mg over 30 min amounted to 103.2 +/- 45.9 microgram/ml, and trough levels at 12 h were 9.6 +/- 3.8 microgram/ml. A dosage of 1,000 mg of meropenem twice a day provides plasma drug levels covering intermediately susceptible microorganisms. Further reductions of the dosage might be appropriate for highly susceptible bacteria or when renal replacement therapies with lower clearances are applied. PMID- 9736576 TI - Influence of human serum on pharmacodynamic properties of an investigational glycopeptide, LY333328, and comparator agents against Staphylococcus aureus. AB - The MICs and MBCs of 15 antibiotics for two strains of Staphylococcus aureus were determined in Mueller-Hinton broth (MHB) and 90% serum-10% MHB. Subsequent experiments established that highly protein-bound antibiotics (>/=80%), such as LY333328, demonstrated higher MICs and MBCs, less killing over an 8-h interval, and shorter postantibiotic effects in 90% serum-10% MHB than in MHB alone. Albumin was demonstrated to be almost solely responsible for changes in the aforementioned pharmacodynamic parameters of LY333328. PMID- 9736577 TI - Superior efficacy of liposomal amphotericin B with prolonged circulation in blood in the treatment of severe candidiasis in leukopenic mice. AB - In leukopenic mice with severe systemic candidiasis, single-dose treatment (5 mg of amphotericin B [AMB]/kg of body weight) with long-circulating polyethylene glycol-coated AMB liposomes (PEG-AMB-LIP) resulted in zero mortality and a significant reduction in the number of viable Candida albicans in the kidney, whereas 70% mortality was seen in mice treated with five daily doses of AmBisome (5 mg of AMB/kg . day). When the first of five daily doses of AmBisome was combined with a single low dose of Fungizone (0.1 mg of AMB/kg), the efficacy was equal to that of PEG-AMB-LIP. PMID- 9736578 TI - Control of Candida albicans murine vaginitis by topical administration of polycarbophil-econazole complex. AB - The complexation of econazole with the mucoadhesive polycarbophil was found to significantly improve the therapeutic benefit of the drug in the topical treatment of experimental vaginal candidiasis in mice, while no difference in the antimycotic activity exerted by econazole and polycarbophil-econazole could be detected in vitro. PMID- 9736579 TI - Activity of liposomal amphotericin B with prolonged circulation in blood versus those of AmBisome and fungizone against intracellular Candida albicans in murine peritoneal macrophages. AB - Activity against intracellular Candida albicans was assessed in C. albicans infected murine peritoneal macrophages exposed to long-circulating pegylated amphotericin B liposomes (PEG-AMB-LIP), AmBisome, or Fungizone. The level of antifungal activity of Fungizone is much higher than that of AmBisome or PEG-AMB LIP, while PEG-AMB-LIP and AmBisome show equivalent activity levels. Previous exposure of uninfected macrophages to PEG-AMB-LIP or AmBisome is advantageous for intracellular antifungal activity. PMID- 9736580 TI - Microbicidal activity of a new silver-containing polymer, SPI-ARGENT II. AB - The survival of three bacterial species and Candida albicans was studied on SPI ARGENT II. The immediate recovery from silver-impregnated polymer and control polymer (1 cm2) was approximately 10(6) to 10(7) microorganisms. After incubation (37 degreesC) and neutralization of silver with horse serum (5%), surviving organisms were recovered. The survival of the microorganisms on the polymer was not found to be influenced by the silver implantation. PMID- 9736581 TI - A Ser315Thr substitution in KatG is predominant in genetically heterogeneous multidrug-resistant Mycobacterium tuberculosis isolates originating from the St. Petersburg area in Russia. AB - Parts of katG and rpoB from 27 Russian Mycobacterium tuberculosis isolates were sequenced to detect mutations causing resistance to isoniazid (INH) and rifampin (RMP), respectively. All 24 INH-resistant isolates had a mutated katG, and 22 of them (91.7%) carried a mutation coding for a Ser315Thr shift. An rpoB mutation was noted for each of the 21 RMP-resistant isolates, with Ser531Leu being the most prevalent change encoded. Only two isolates had identical IS6110 fingerprints. PMID- 9736582 TI - Susceptibilities of Candida species isolated from the lower gastrointestinal tracts of high-risk patients to the new semisynthetic echinocandin LY303366 and other antifungal agents. AB - Fifty-two percent of stool specimens collected from 1,200 high-risk patients were colonized with yeasts, primarily Candida albicans (53. 6%) and Candida glabrata (35.7%). Susceptibilities to all antifungal agents tested, including LY303366, were similar to those reported previously for Candida species isolated from blood. PMID- 9736583 TI - Synergism between poly-(1-6)-beta-D-glucopyranosyl-(1-3)-beta-D-glucopyranose glucan and cefazolin in prophylaxis of staphylococcal wound infection in a guinea pig model. AB - To determine whether the infection-preventing capability of the neutrophil activating agent poly-(1-6)-beta-D-glucopyranosyl-(1-3)-beta-D-glucopyranose glucan (PGG-glucan) can be enhanced with antibiotic prophylaxis, we administered PGG-glucan and cefazolin, alone and in combination, to guinea pigs inoculated with isolates of staphylococci. Guinea pigs receiving both PGG-glucan and cefazolin had 50% infective doses that were 8- to 20-fold higher than those obtained with cefazolin alone and 100- to 200-fold higher than those obtained with PGG-glucan alone. PGG-glucan and cefazolin are synergistic in their ability to prevent staphylococcal wound infection. PMID- 9736585 TI - Antimalarial activities of polyhydroxyphenyl and hydroxamic acid derivatives. AB - Several known mammalian ribonucleotide reductase inhibitors featuring a polyhydroxyphenyl and/or hydroxamate moiety as the active group were screened for potency in inhibiting growth of the malaria parasite Plasmodium falciparum. Compounds containing a 2,3- or 3,4-dihydroxyphenyl group as well as benzohydroxamate appear to be the most effective inhibitors of the malaria parasite. PMID- 9736584 TI - In vitro activity of the new glycopeptide LY333328 against multiply resistant gram-positive clinical isolates. AB - The in vitro activity of LY333328 was compared with those of vancomycin and teicoplanin against 425 gram-positive clinical isolates, including a variety of multiply resistant strains. LY333328 at 16, respectively. Gatifloxacin MICs were similar to those of trovafloxacin in all organism groups. PMID- 9736588 TI - AHA journals lead with definitive new online site. PMID- 9736587 TI - Differences in the occurrence of two base pair variants of Tn1546 from vancomycin resistant enterococci from humans, pigs, and poultry. PMID- 9736589 TI - Feasibility of combined percutaneous transluminal angioplasty and minimally invasive direct coronary artery bypass in patients with multivessel coronary artery disease. AB - BACKGROUND: Angioplasty has become an accepted treatment of patients with coronary artery disease and is now commonly used to treat patients with multivessel disease. The major disadvantage of angioplasty has been restenosis requiring repeat interventions with resultant loss of initial cost savings. Compared with the right and the circumflex coronary arteries, the left anterior descending artery (LAD) has been more adversely affected by restenosis. Recently, minimally invasive direct coronary artery bypass (MIDCAB) to the LAD through a small left anterior thoracotomy using the left internal mammary artery has been performed in some centers with excellent early results and with reduced costs compared with standard bypass surgery. METHODS AND RESULTS: We retrospectively reviewed the first 31 consecutive patients treated in our institution with integrated coronary revascularization (ICR): MIDCAB to the LAD combined with PTCA of the other diseased vessels in patients with multivessel disease. Postoperative angiography in 84% of patients revealed a patent anastomosis and normal flow in the graft and bypassed vessel. Thirty-eight (97%) of 39 vessels were successfully treated percutaneously. At a mean follow-up of 7 months, all patients are currently asymptomatic. There have been 2 adverse clinical events, both related to angioplasty and not to MIDCAB. The average length of stay at the hospital after MIDCAB was 2.79+/-1.05 days. CONCLUSIONS: These preliminary results with ICR are encouraging and suggest that a randomized, prospective clinical trial comparing ICR with standard coronary artery bypass surgery for the revascularization of symptomatic patients with multivessel disease involving the LAD is warranted. PMID- 9736590 TI - Presence of tissue factor pathway inhibitor in human atherosclerotic plaques is associated with reduced tissue factor activity. AB - BACKGROUND: Plaque disruption and exposure of subendothelial procoagulants such as tissue factor (TF) to circulating factor VII/VIIa (FVII/VIIa) lead to intravascular thrombosis. Tissue factor pathway inhibitor (TFPI) is an endogenous inhibitor of TF-induced coagulation that binds to factor Xa and the TF-FVIIa catalytic complex in a two-step process. The aim of this study was to determine the expression of TFPI within human atherosclerotic plaque and its role in modulation of TF activity. METHODS AND RESULTS: We measured the level of TFPI antigen in human carotid plaque and determined the relationship between TFPI and TF activity within plaque. Furthermore, we examined the biological activity and immunolocalization patterns of TFPI within carotid plaque. TFPI was detectable (TFPI+ group) in 22 of 34 specimens (mean+/-SEM, 404. 4+/-91.8 pg/mg) and undetectable (TFPI- group) in 12 of 34 specimens. In the TFPI- group, normalized TF activity was significantly greater than that in the TFPI+ group (0.28+/-0.04 vs 0.14+/-0.02 U/pg, P=0.002). Furthermore, neutralization of TFPI activity using a polyclonal antibody resulted in an 8-fold increase in TF activity in the TFPI+ group (P=0.001) but had no effect in the TFPI- group. Immunostaining for TFPI showed localization to endothelial cells, vascular smooth muscle cells within the fibrous cap region of the plaque, and macrophages within the shoulder region of the plaque. CONCLUSIONS: Taken together, these data suggest that biologically active TFPI is present within human atherosclerotic plaque and is associated with attenuated TF activity. PMID- 9736591 TI - Myocardial infarction and use of low-dose oral contraceptives: a pooled analysis of 2 US studies. AB - BACKGROUND: Population-based case-control studies to assess the relationship of low-dose oral contraceptive (OC) use with myocardial infarction (MI) were performed at 2 sites in the United States (California and Washington state). The purpose of the present study was to estimate risk of MI in relation to use of low dose OCs in a pooled analysis combining results from the 2 sites. METHODS AND RESULTS: The study included as cases women aged 18 to 44 years with incident MI who had no prior history of ischemic heart disease or cerebrovascular disease. Women in the case and control groups were interviewed in person regarding OC use and cardiovascular risk factors. The analysis included 271 MI cases and 993 controls. Compared with noncurrent users, the adjusted pooled odds ratio for MI in current OC users was 0.94 (95% CI, 0.44, 2.20) after adjustment for major risk factors and sociodemographic factors. Compared with never users, the adjusted pooled odds ratio for MI was 0.56 (0.21, 1.49) in current OC users and 0.54 (0.31, 0.95) in past OC users. Among past OC users, duration and recency of use were unrelated to MI risk as was current hormone replacement therapy. There was no evidence of interaction between OC use and age, presence of cardiovascular risk factors (hypercholesterolemia, hypertension, diabetes), obesity, or smoking. CONCLUSIONS: We conclude that low-dose OCs as used in these populations are safe with respect to risk of MI in women. PMID- 9736592 TI - Long-term oral anticoagulant therapy in patients with unstable angina or suspected non-Q-wave myocardial infarction: organization to assess strategies for ischemic syndromes (OASIS) pilot study results. AB - BACKGROUND: Patients with acute ischemic syndromes (AIS) suffer high rates of recurrent ischemic events despite aspirin treatment. Long-term therapy with oral anticoagulants in addition to aspirin may reduce this risk. We studied the effects of long-term warfarin at 2 intensities in patients with AIS without ST elevation in 2 consecutive randomized controlled studies. METHODS AND RESULTS: In phase 1, after the cessation of 3 days of intravenous antithrombotic therapy, 309 patients were randomized to receive fixed low-dose (3 mg/d) warfarin for 6 months that produced a mean international normalized ratio (INR) of 1.5+/-0.6 or to standard therapy. Eighty-seven percent of patients received aspirin in both groups. The rates of cardiovascular (CV) death, new myocardial infarction (MI), and refractory angina at 6 months were 6.5% in the warfarin group and 3.9% in the standard therapy group (relative risk [RR], 1. 66; 95% CI, 0.62 to 4.44; P=0.31). The rates of death, new MI, and stroke were 6.5% in the warfarin group and 2.6% in the standard therapy group (RR, 2.48; 95% CI, 0.80 to 7.75; P=0.10). The overall rate of rehospitalization for unstable angina was 21% and did not differ significantly between the groups. Four patients in the warfarin group (2.6%) and none in the control group experienced a major bleed (RR, 2.48; 95% CI, 0.80 to 7.75), and there was a significant excess of minor bleeds in the warfarin group (14.2% versus 2.6%; RR, 5.46; 95% CI, 1.93 to 15.5; P=0.001). In phase 2, the protocol was modified, and 197 patients were randomized <48 hours from the onset of symptoms to receive warfarin at an adjusted dose that produced a mean INR of 2.3+/-0.6 or standard therapy for 3 months. Eighty-five percent received aspirin in both groups. The rates of CV death, new MI, and refractory angina at 3 months were 5. 1% in the warfarin group and 12.1% in the standard group (RR, 0.42; 95% CI, 0.15 to 1.15; P=0.08). The rates of all death, new MI, and stroke were 5.1% in the warfarin group and 13.1% in the standard therapy group (RR, 0.39; 95% CI, 0.14 to 1.05; P=0.05). Significantly fewer patients were rehospitalized for unstable angina in the warfarin group than in the control group (7.1% and 17.2%, respectively; RR, 0.42; 95% CI, 0.18 to 0.96; P=0.03). Two patients in the warfarin group and 1 in the control group experienced a major bleed, and there was a significant excess of minor bleeds in the warfarin group (28.6% versus 12.1%; RR, 2.36; 95% CI, 1.37 to 4.36; P=0.004). CONCLUSIONS: Long-term treatment with moderate-intensity warfarin (INR, 2.0 to 2.5) plus aspirin but not low intensity warfarin (INR, 1.5) plus aspirin appears to reduce the rate of recurrent ischemic events in patients with AIS without ST elevation. PMID- 9736593 TI - Altered cardiovascular variability in obstructive sleep apnea. AB - BACKGROUND: Altered cardiovascular variability is a prognostic indicator for cardiovascular events. Patients with obstructive sleep apnea (OSA) are at an increased risk for cardiovascular disease. We tested the hypothesis that OSA is accompanied by alterations in cardiovascular variability, even in the absence of overt cardiovascular disease. METHODS AND RESULTS: Spectral analysis of variability of muscle sympathetic nerve activity, RR interval, and blood pressure were obtained during undisturbed supine rest in 15 patients with moderate-to severe OSA, 18 patients with mild OSA, and 16 healthy control subjects in whom sleep disordered breathing was excluded by complete overnight polysomnography. Patients with OSA were newly diagnosed, never treated for OSA, and free of any other known diseases. Patients with moderate-to-severe OSA had shorter RR intervals (793+/-27 ms) and increased sympathetic burst frequency (49+/-4 bursts/min) compared with control subjects (947+/-42 ms; 24+/-3 bursts/min; P=0.008 and P<0.001, respectively). In these patients, total variance of RR was reduced (P=0.01) and spectral analysis of RR variability showed an increase in low frequency normalized units, a decrease in high frequency normalized units, and an increase in the ratio of low to high frequency (all P<0.05). Even though blood pressure was similar to that of the control subjects, blood pressure variance in patients with moderate-to-severe OSA was more than double the variance in control subjects (P=0.01). Patients with mild OSA also had a reduction in RR variance (P=0.02) in the absence of any significant difference in absolute RR interval. For all patients with OSA, linear regression showed a positive correlation (r=0.40; P=0.02) between sleep apnea severity and blood pressure variance. CONCLUSIONS: Cardiovascular variability is altered in patients with OSA. This alteration is evident even in the absence of hypertension, heart failure, or other disease states and may be linked to the severity of OSA. Abnormalities in cardiovascular variability may be implicated in the subsequent development of overt cardiovascular disease in patients with OSA. PMID- 9736594 TI - Prognostic value of myocardial viability in medically treated patients with global left ventricular dysfunction early after an acute uncomplicated myocardial infarction: a dobutamine stress echocardiographic study. AB - BACKGROUND: Residual viable myocardium identified by dobutamine stress after myocardial infarction may act as an unstable substrate for further events such as subsequent angina and reinfarction. However, in patients with severe global left ventricular dysfunction, viability might be protective rather than detrimental. The aim of this study was to assess the impact on survival of echocardiographically detected viability in medically treated patients with global left ventricular dysfunction evaluated after acute uncomplicated myocardial infarction. METHODS AND RESULTS: The data bank of the large-scale, prospective, multicenter, observational Echo Dobutamine International Cooperative (EDIC) study was interrogated to select 314 medically treated patients (271 men; age, 58+/-9 years) who underwent low-dose (1.6). Patients were followed up for 9+/-7 months. Low dose dobutamine stress echocardiography identified myocardial viability in 130 patients (52%). Dobutamine-atropine stress echocardiography was positive for ischemia in 148 patients (47%) and negative in 166 patients (53%). During the follow-up, there were 12 cardiac deaths (3.8% of the total population). With the use of Cox proportional hazards model, delta low-dose WMSI (the variation between rest WMSI and low-dose WMSI) was shown to exert a protective effect by reducing cardiac death by 0.8 for each decrease in WMSI at low-dose dobutamine (coefficient, -0.2; hazard ratio, 0.8; P<0.03); WMSI at peak stress was the best predictor of cardiac death in this set of patients (hazard ratio, 14.9; P<0.0018). CONCLUSIONS: In medically treated patients with severe global left ventricular dysfunction early after acute uncomplicated myocardial infarction, the presence of myocardial viability identified as inotropic reserve after low dose dobutamine is associated with a higher probability of survival. The higher the number of segments showing improvement of function, the better the impact is of myocardial viability on survival. The presence of inducible ischemia in this set of patients is the best predictor of cardiac death. PMID- 9736595 TI - Abciximab (ReoPro, chimeric 7E3 Fab) demonstrates equivalent affinity and functional blockade of glycoprotein IIb/IIIa and alpha(v)beta3 integrins. AB - BACKGROUND: Large, randomized, and blinded clinical trials (EPIC, EPILOG, and CAPTURE) have demonstrated that abciximab (ReoPro, chimeric 7E3 Fab) markedly reduces thrombotic events associated with percutaneous transluminal coronary interventions. The marked early benefits at 30 days were sustained at 6 months and 3 years. Initially developed because of its efficacy in blocking GP IIb/IIIa (alphaIIb/beta3) receptors on platelets, abciximab also binds with equivalent affinity to alpha(v)beta3. METHODS AND RESULTS: This study presents a detailed characterization of the alphavss3 interaction, including the ability of abciximab to (1) bind with comparable affinity to alpha(v)beta3 and GP IIb/IIIa, (2) inhibit alpha(v)beta3 and GP IIb/IIIa-mediated cell adhesion in vitro with IC50 values approximating binding KD values, and (3) redistribute between GP IIb/IIIa and alpha(v)beta3 integrins in vitro. CONCLUSIONS: As an antagonist of not only GP IIb/IIIa but also alpha(v)beta3, abciximab may provide additional clinical benefit in preventing alpha(v)beta3-mediated effects such as thrombin generation, clot retraction, or smooth muscle cell migration and proliferation. Abciximab binds with equivalent affinity to both GP IIb/IIIa and alphavss3 and redistributes between the 2 integrin receptors in vitro. Abciximab has been previously shown to circulate on platelets for up to 2 weeks. Taken together, these findings suggest that abciximab may have the ability to inhibit both GP IIb/IIIa and alpha(v)beta3 for extended periods. PMID- 9736596 TI - Antibodies to endothelial cells in borderline hypertension. AB - BACKGROUND: Antibodies to endothelial cells (aECs) and to cardiolipin (aCLs) are implicated in autoimmune diseases like systemic lupus erythematosus vasculitis. Beta2-Glycoprotein 1 (beta2GP1) is a cofactor for aCLs. The present study investigated the possible role of aECs, aCLs, and abeta2GP1 in borderline hypertension. METHODS AND RESULTS: Seventy-three men with borderline hypertension (BHT) and 73 age-matched normotensive (NT) men (diastolic blood pressure, 85 to 94 and <80 mm Hg, respectively) were recruited from a population screening program. Antibody levels were determined by ELISA. Presence of carotid atherosclerosis was determined by B-mode ultrasonography, and 29 individuals had atherosclerotic plaques. BHT men had significantly higher aEC and abeta2GP1 levels of IgG class than NT control subjects (P=0.029 and P=0.0001, respectively). aEC levels of IgM class were higher in BHT (P=0.012), but not abeta2GP1 levels. There was no correlation between aCL levels and BHT. Individuals with atherosclerotic plaques had significantly higher aEC levels of both IgG (P=0.042) and IgM subclasses (P=0.018) than those without plaques, but no difference was found in aCL and abeta2GP1 levels. Endothelin and aECs of IgM class were significantly associated. CONCLUSIONS: We demonstrate the first evidence of a significant elevation of aEC and abeta2GP1 levels in borderline hypertension. These findings provide a new link between hypertension and atherosclerosis and indicate that humoral immune reactions to the endothelium may play an important role in both conditions. PMID- 9736597 TI - Predictors of early- and late-onset supraventricular tachyarrhythmias after Fontan operation. AB - BACKGROUND: The objectives of our study were to determine the frequency of supraventricular tachyarrhythmias (SVTAs) among modifications of the Fontan operation and identify risk factors for developing SVTA. METHODS AND RESULTS: The population consisted of all patients who had any modification of the Fontan operation at the Mayo Clinic between 1985 and 1993. Clinically significant SVTAs were those requiring initiation or change of antiarrhythmic treatment, and they were divided into early SVTAs (<30 days after the operation) and late SVTAs (>/=30 days after the operation). Clinical histories were reviewed, and health status questionnaires were sent. Four hundred ninety-nine patients had various modifications of the Fontan operation. Frequency of early SVTA was 15%. Risk factors identified by multivariate analysis for early SVTA were AV valve regurgitation, abnormal AV valve, and preoperative SVTA. Frequency of late SVTA was 6% by 1 year, 12% by 3 years, and 17% by 5 years. Risk factors for late SVTA were age at operation (<3 or >/=10 years) and systemic AV valve replacement. By univariate and multivariate analysis, the type of Fontan operation was not a significant risk factor for late SVTA when all 6 modifications were considered. However, when we analyzed the frequency of late SVTA for the 2 recently used modifications, we found a lower frequency of late SVTA in patients with atriopulmonary connection with lateral tunnel compared with those with total cavopulmonary connection. CONCLUSIONS: Postoperative SVTA continues to be a significant problem. Risk factors for SVTA are AV valve regurgitation, abnormal AV valve, preoperative SVTA, and age at operation. Frequency of SVTA does not appear to be related to type of Fontan procedure except for slightly lower frequency in patients with atriopulmonary connection with lateral tunnel compared with those with total cavopulmonary connection. PMID- 9736599 TI - Preliminary animal and clinical experiences using an electromechanical endocardial mapping procedure to distinguish infarcted from healthy myocardium. AB - BACKGROUND: A catheter-based left ventricular (LV) endocardial mapping procedure using electromagnetic field energy for positioning of the catheter tip was designed to acquire simultaneous measurements of endocardial voltage potentials and myocardial contractility. We investigated such a mapping system to distinguish between infarcted and normal myocardium in an animal infarction model and in patients with coronary artery disease. METHODS AND RESULTS: Measurements of LV endocardial unipolar (UP) and bipolar (BP) voltages and local endocardial shortening were derived from dogs at baseline (n=12), at 24 hours (n=6), and at 3 weeks (n=6) after occlusion of the left anterior descending coronary artery. Also, 12 patients with prior myocardial infarction (MI) and 12 control patients underwent the LV endocardial mapping study for assessment of electromechanical function in infarcted versus healthy myocardial regions. In the canine model, a significant decrease in voltage potentials was noted in the MI zone at 24 hours (UP, 42. 8+/-9.6 to 29.1+/-12.2 mV, P=0.007; BP, 11.6+/-2.3 to 4.9+/-1.2 mV, P<0.0001) and at 3 weeks (UP, 41.0+/-8.9 to 13.9+/-3.9 mV, P<0.0001; BP, 11.2+/ 2.8 to 2.4+/-0.4 mV, P<0.0001). No change in voltage was noted in zones remote from MI. In patients with prior MI, the average voltage was 7.2+/-2.7 mV (UP)/1.4+/-0.7 mV (BP) in MI regions, 17.8+/-4.6 mV (UP)/4.5+/-1.1 mV (BP) in healthy zones remote from MI, and 19.7+/-4.4 mV (UP)/5.8+/-1.0 mV (BP) in control patients without prior MI (P<0.001 for MI values versus remote zones or control patients). In the canine model and patients, local endocardial shortening was significantly impaired in MI zones compared with controls. CONCLUSIONS: These preliminary data suggest that infarcted myocardium could be accurately diagnosed and distinguished from healthy myocardium by a reduction in both electrical voltage and mechanical activity. Such a diagnostic electromechanical mapping study might be clinically useful for accurate assessment of myocardial function and viability. PMID- 9736598 TI - Induction of early atherosclerosis in LDL-receptor-deficient mice immunized with beta2-glycoprotein I. AB - BACKGROUND: Immunization with beta2-glycoprotein I (beta2GPI), the probable target of autoimmune anticardiolipin antibodies, results in experimental antiphospholipid syndrome in different mouse strains. The present study was undertaken to evaluate the effect of beta2GPI immunization on the progression of atherosclerosis. METHODS AND RESULTS: In the first experiment, 3 groups of LDL receptor-deficient (LDL-RD) mice (n=15 per group) were immunized with either beta2GPI or ovalbumin or were not immunized and were fed a chow diet for 12 weeks. In a second experiment, 3 groups of LDL-RD mice (n=10 per group) were immunized similarly and fed an atherogenic diet for 6 weeks. All beta2GPI immunized mice developed high titers of anti-beta2GPI antibodies as well as a specific lymph node proliferation to beta2GPI. The average cholesterol levels did not differ between the mice fed similar diets, regardless of the immunization protocol. Atherosclerosis was enhanced in the beta2GPI-immunized mice (mean aortic lesion, 26 000+/-5700 microm2) in comparison with their ovalbumin immunized (mean, 3000+/-1099 microm2; P<0.01) and nonimmunized (mean, 2250+/-700 microm2; P<0.01) littermates. The average lesion size in the beta2GPI-immunized mice fed an atherogenic diet (mean, 98 000+/-8305 microm2) was larger than the ovalbumin-immunized mice (mean, 81 250+/-12 933 microm2; P=NS) or the nonimmunized controls (mean, 75 625+/-7281 microm2; P=NS). The atherosclerotic plaques in the beta2GPI-immunized mice appeared to be more mature, and denser infiltration of CD4 lymphocytes was present in the subendothelium of the aortic sinuses from this group of mice. CONCLUSIONS: The results of the present study provide the first direct evidence for the proatherogenic effect of ss2GPI immunization and establish a new model for immune-mediated atherosclerosis. PMID- 9736600 TI - Enhanced susceptibility for acquired torsade de pointes arrhythmias in the dog with chronic, complete AV block is related to cardiac hypertrophy and electrical remodeling. AB - BACKGROUND: Chronic, complete AV block (CAVB) in the dog leads to ventricular hypertrophy, which has been described as an independent risk factor for arrhythmias. In this model, we examined (1) whether the short- and long-term electrical adaptations predispose to acquired torsade de pointes arrhythmias (TdP) and (2) the nature of the structural and functional adaptations involved. METHODS AND RESULTS: We determined (1) endocardial right (RV) and left (LV) ventricular APD, DeltaAPD (LV APD-RV APD), presence of EADs at 0 weeks (acute: AAVB), and CAVB (6 weeks) and inducibility of TdP by pacing and d-sotalol (n=10); (2) steady-state and dynamic LV hemodynamics at 0 and 6 weeks (n=6); (3) plasma neurohumoral levels in time (n=7); (4) structural parameters of the LV and RV of CAVB dogs (n=6) compared with sinus rhythm (SR) dogs (n=6); and (5) expression of ventricular mRNA atrial natriuretic factor (ANF) in CAVB (n=4) and SR (n=4) dogs. Compared with AAVB, CAVB led to nonhomogeneous prolongation of LV and RV APD and different sensitivity for d-sotalol, leading to EADs (4 of 14 versus 9 of 18, P<0.05), increased DeltaAPD (45+/-30 versus 125+/-60 ms, P<0.05), and induction of TdP in most dogs (0% versus 60%, P<0.05). CAVB led to biventricular hypertrophy, whereas LV function was similar in AAVB and CAVB. The neurohumoral levels were transiently elevated. The LV and RV collagen and the capillary/fiber ratio remained normal, whereas ventricular ANF mRNA was not detectable. CONCLUSIONS: The electrical remodeling occurring after CAVB predisposes the heart to acquired TdP, whereas the structural changes (hypertrophy) are successfully aimed at maintaining cardiac function. PMID- 9736601 TI - Cellular basis of biventricular hypertrophy and arrhythmogenesis in dogs with chronic complete atrioventricular block and acquired torsade de pointes. AB - BACKGROUND: In the dog with chronic complete atrioventricular block (AVB), torsade de pointes arrhythmias (TdP) can be induced reproducibly by class III antiarrhythmic agents. In vivo studies reveal important electrophysiological alterations of the heart at 5 weeks of AVB, resulting in increased proarrhythmia. Autopsy studies indicate the presence of biventricular hypertrophy. In this study, the cellular basis of proarrhythmia and hypertrophy in chronic AVB was investigated. METHODS AND RESULTS: From chronic-AVB dogs with increased heart weights and TdP, left midmyocardial and right ventricular myocytes were isolated by enzymatic dispersion. These myocytes were significantly larger than sinus rhythm (SR) controls. In chronic AVB, the action potential spike-and-dome configuration was preserved. However, the action potential duration (APD) at 95% and 50% of repolarization of the left midmyocardium was significantly larger in chronic AVB than in SR, with little change in the right ventricle, causing enhanced interventricular dispersion of repolarization at slow pacing rates. Treatment with the class III agent almokalant increased the APD to a much larger extent in chronic-AVB than in SR myocytes and resulted in a higher incidence of early afterdepolarizations (EADs). EADs had their takeoff potential between -35 and 0 mV. There was no evidence that spontaneous sarcoplasmic reticulum Ca2+ release underlies these EADs. CONCLUSIONS: In the dog, chronic AVB leads to hypertrophy of both right and left ventricular myocytes. The repolarization abnormalities predisposing for class III-dependent TdP in vivo are the results of cellular electrophysiological remodeling. PMID- 9736602 TI - Aortic valve stenosis in alkaptonuria. Images in cardiovascular medicine. PMID- 9736603 TI - Pulmonary arterial obstruction due to a huge sinus venosus remnant. Images in cardiovascular medicine. PMID- 9736604 TI - Protein folding and misfolding inside and outside the cell. AB - The workshop was held at St Catherine's College, Oxford, from March 25-28, 1998, and attracted participants from 32 nations. Protein folding is one of the most important processes in biology since it adds functional flesh to the bare bones of genes, but it has traditionally been studied by people separated both intellectually and physically because they are training in different disciplines. The aim of the meeting was to bring together chemists and structural biologists studying how pure, denatured proteins refold spontaneously in the test tube, with biochemists and cell biologists who are concerned with how proteins fold inside living cells and medical scientists interested in the diseases that result when this process goes wrong. In this report we concentrate on general concepts and themes rather than on detailing every contribution. PMID- 9736605 TI - Phospholipid-assisted protein folding: phosphatidylethanolamine is required at a late step of the conformational maturation of the polytopic membrane protein lactose permease. AB - Previously we presented evidence that phosphatidylethanolamine (PE) acts as a molecular chaperone in the folding of the polytopic membrane protein lactose permease (LacY) of Escherichia coli. Here we provide more definitive evidence supporting the chaperone properties of PE. Membrane insertion of LacY prevents its irreversible aggregation, and PE participates in a late step of conformational maturation. The temporal requirement for PE was demonstrated in vitro using a coupled translation-membrane insertion assay that allowed the separation of membrane insertion from phospholipid-assisted folding. LacY was folded properly, as assessed by recognition with conformation-specific monoclonal antibodies, when synthesized in the presence of PE-containing inside-out membrane vesicles (IOVs) or in the presence of IOVs initially lacking PE but supplemented with PE synthesized in vitro either co- or post-translationally. The presence of IOVs lacking PE and containing anionic phospholipids or no addition of IOVs resulted in misfolded or aggregated LacY, respectively. Therefore, critical folding steps occur after membrane insertion dependent on the interaction of LacY with PE to prevent illicit interactions which lead to misfolding of LacY. PMID- 9736606 TI - All-trans retinol, vitamin D and other hydrophobic compounds bind in the axial pore of the five-stranded coiled-coil domain of cartilage oligomeric matrix protein. AB - The potential storage and delivery function of cartilage oligomeric matrix protein (COMP) for cell signaling molecules was explored by binding hydrophobic compounds to the recombinant five-stranded coiled-coil domain of COMP. Complex formation with benzene, cyclohexane, vitamin D3 and elaidic acid was demonstrated through increases in denaturation temperatures of 2-10 degreesC. For all-trans retinol and all-trans retinoic acid, an equilibrium dissociation constant KD = 0.6 microM was evaluated by fluorescence titration. Binding of benzene and all trans retinol into the hydrophobic axial pore of the COMP coiled-coil domain was proven by the X-ray crystal structures of the corresponding complexes at 0.25 and 0.27 nm resolution, respectively. Benzene binds with its plane perpendicular to the pore axis. The binding site is between the two internal rings formed by Leu37 and Thr40 pointing into the pore of the COMP coiled-coil domain. The retinol beta ionone ring is positioned in a hydrophobic environment near Thr40, and the 1.1 nm long isoprene tail follows a completely hydrophobic region of the pore. Its terminal hydroxyl group complexes with a ring of the five side chains of Gln54. A mutant in which Gln54 is replaced by Ile binds all-trans retinol with affinity similar to the wild-type, demonstrating that hydrophobic interactions are predominant. PMID- 9736607 TI - Crystal structure of the amphiphysin-2 SH3 domain and its role in the prevention of dynamin ring formation. AB - The amphiphysins are brain-enriched proteins, implicated in clathrin-mediated endocytosis, that interact with dynamin through their SH3 domains. To elucidate the nature of this interaction, we have solved the crystal structure of the amphiphysin-2 (Amph2) SH3 domain to 2.2 A. The structure possesses several notable features, including an extensive patch of negative electrostatic potential covering a large portion of its dynamin binding site. This patch accounts for the specific requirement of amphiphysin for two arginines in the proline-rich binding motif to which it binds on dynamin. We demonstrate that the interaction of dynamin with amphiphysin SH3 domains, unlike that with SH3 domains of Grb2 or spectrin, prevents dynamin self-assembly into rings. Deletion of a unique insert in the n-Src loop of Amph2 SH3, a loop adjacent to the dynamin binding site, significantly reduces this effect. Conversely, replacing the n-Src loop of the N-terminal SH3 domain of Grb2 with that of Amph2 causes it to favour dynamin ring disassembly. Transferrin uptake assays show that shortening the n Src loop of Amph2 SH3 reduces the ability of this domain to inhibit endocytosis in vivo. Our data suggest that amphiphysin SH3 domains are important regulators of the multimerization cycle of dynamin in endocytosis. PMID- 9736608 TI - A nuclear-encoded protein of prokaryotic origin is essential for the stability of photosystem II in Arabidopsis thaliana. AB - To understand the regulatory mechanisms underlying the biogenesis of photosystem II (PSII) we have characterized the nuclear mutant hcf136 of Arabidopsis thaliana and isolated the affected gene. The mutant is devoid of any photosystem II activity, and none of the nuclear- and plastome-encoded subunits of this photosystem accumulate to significant levels. Protein labelling studies in the presence of cycloheximide showed that the plastome-encoded PSII subunits are synthesized but are not stable. The HCF136 gene was isolated by virtue of its T DNA tag, and its identity was confirmed by complementation of homozygous hcf136 seedlings. Immunoblot analysis of fractionated chloroplasts showed that the HCF136 protein is a lumenal protein, found only in stromal thylakoid lamellae. The HCF136 protein is produced already in dark-grown seedlings and its levels do not increase dramatically during light-induced greening. This accumulation profile confirms the mutational data by showing that the HCF136 protein must be present when PSII complexes are made. HCF136 homologues are found in the cyanobacterium Synechocystis species PCC6803 (slr2034) and the cyanelle genome of Cyanophora paradoxa (ORF333), but are lacking in the plastomes of chlorophytes and metaphytes as well as from those of rhodo- and chromophytes. We conclude that HCF136 encodes a stability and/or assembly factor of PSII which dates back to the cyanobacterial-like endosymbiont that led to the plastids of the present photosynthetic eukaryotes. PMID- 9736609 TI - The Golgi apparatus is an inositol 1,4,5-trisphosphate-sensitive Ca2+ store, with functional properties distinct from those of the endoplasmic reticulum. AB - In the past few years, intracellular organelles, such as the endoplasmic reticulum, the nucleus and the mitochondria, have emerged as key determinants in the generation and transduction of Ca2+ signals of high spatio-temporal complexity. Little is known about the Golgi apparatus, despite the fact that Ca2+ within its lumen controls essential processes, such as protein processing and sorting. We report the direct monitoring of the [Ca2+] in the Golgi lumen ([Ca2+]Golgi) of living HeLa cells, using a specifically targeted Ca2+-sensitive photoprotein. With this probe, we show that, in resting cells, [Ca2+]Golgi is approximately 0.3 mM and that Ca2+ accumulation by the Golgi has properties distinct from those of the endoplasmic reticulum (as inferred by the sensitivity to specific inhibitors). Upon stimulation with histamine, an agonist coupled to the generation of inositol 1,4,5-trisphosphate (IP3), a large, rapid decrease in [Ca2+]Golgi is observed. The Golgi apparatus can thus be regarded as a bona fide IP3-sensitive intracellular Ca2+ store, a notion with major implications for the control of organelle function, as well as for the generation of local cytosolic Ca2+ signals. PMID- 9736610 TI - Severe B cell deficiency and disrupted splenic architecture in transgenic mice expressing the E41K mutated form of Bruton's tyrosine kinase. AB - To identify B-cell signaling pathways activated by Bruton's tyrosine kinase (Btk) in vivo, we generated transgenic mice in which Btk expression is driven by the MHC class II Ea gene locus control region. Btk overexpression did not have significant adverse effects on B cell function, and essentially corrected the X linked immunodeficiency (xid) phenotype in Btk- mice. In contrast, expression of a constitutively activated form of Btk carrying the E41K gain-of-function mutation resulted in a B cell defect that was more severe than xid. The mice showed a marked reduction of the B cell compartment in spleen, lymph nodes, peripheral blood and peritoneal cavity. The levels in the serum of most immunoglobulin subclasses decreased with age, and B cell responses to both T cell independent type II and T cell-dependent antigens were essentially absent. Expression of the E41K Btk mutant enhanced blast formation of splenic B cells in vitro in response to anti-IgM stimulation. Furthermore, the mice manifested a disorganization of B cell areas and marginal zones in the spleen. Our findings demonstrate that expression of constitutively activated Btk blocks the development of follicular recirculating B cells. PMID- 9736611 TI - Transformation of hematopoietic cell lines to growth-factor independence and induction of a fatal myelo- and lymphoproliferative disease in mice by retrovirally transduced TEL/JAK2 fusion genes. AB - Recent reports have demonstrated fusion of the TEL gene on 12p13 to the JAK2 gene on 9p24 in human leukemias. Three variants have been identified that fuse the TEL pointed (PNT) domain to (i) the JAK2 JH1-kinase domain, (ii) part of and (iii) all of the JH2 pseudokinase domain. We report that all of the human TEL/JAK2 variants, and a human/mouse chimeric hTEL/mJAK2(JH1) fusion gene, transform the interleukin-3 (IL-3)-dependent murine hematopoietic cell line Ba/F3 to IL-3 independent growth. Transformation requires both the TEL PNT domain and JAK2 kinase activity. Furthermore, all TEL/JAK2 variants strongly activated STAT 5 by phosphotyrosine Western blots and by electrophoretic mobility shift assays (EMSA). Mice (n = 40) transplanted with bone marrow infected with the MSCV retrovirus containing either the hTEL/mJAK2(JH1) fusion or its human counterpart developed a fatal mixed myeloproliferative and T-cell lymphoproliferative disorder with a latency of 2-10 weeks. In contrast, mice transplanted with a TEL/JAK2 mutant lacking the TEL PNT domain (n = 10) or a kinase-inactive TEL/JAK2(JH1) mutant (n = 10) did not develop the disease. We conclude that all human TEL/JAK2 fusion variants are oncoproteins in vitro that strongly activate STAT 5, and cause lethal myelo- and lymphoproliferative syndromes in murine bone marrow transplant models of leukemia. PMID- 9736613 TI - Evidence implicating Gfi-1 and Pim-1 in pre-T-cell differentiation steps associated with beta-selection. AB - After rearrangement of the T-cell receptor (TCR) beta-locus, early CD4(-)/CD8(-) double negative (DN) thymic T-cells undergo a process termed 'beta-selection' that allows the preferential expansion of cells with a functional TCR beta-chain. This process leads to the formation of a rapidly cycling subset of DN cells that subsequently develop into CD4(+)/CD8(+) double positive (DP) cells. Using transgenic mice that constitutively express the zinc finger protein Gfi-1 and the serine/threonine kinase Pim-1, we found that the levels of both proteins are important for the correct development of DP cells from DN precursors at the stage where 'beta-selection' occurs. Analysis of the CD25(+)/CD44(-,lo) DN subpopulation from these animals revealed that Gfi-1 inhibits and Pim-1 promotes the development of larger beta-selected cycling cells ('L subset') from smaller resting cells ('E subset') within this subpopulation. We conclude from our data that both proteins, Pim-1 and Gfi-1, participate in the regulation of beta selection-associated pre-T-cell differentiation in opposite directions and that the ratio of both proteins is important for pre-T-cells to pass the 'E' to 'L' transition correctly during beta-selection. PMID- 9736612 TI - Engagement of T cell receptor triggers its recruitment to low-density detergent insoluble membrane domains. AB - T-cell receptors (TCRs) upon binding to peptide-MHC ligands transduce signals in T lymphocytes. Tyrosine phosphorylations in the cytoplasmic domains of the CD3 (gammadeltaepsilon) and zeta subunits of the TCR complex by Src family kinases initiate the signaling cascades via docking and activation of ZAP-70 kinase and other signaling components. We examined the role of the low-density detergent insoluble membranes (DIMs) in TCR signaling. Using mouse thymocytes as a model, we characterized the structural organization of DIMs in detail. We then demonstrated that TCR engagement triggered an immediate increase in the amount of TCR/CD3 present in DIMs, which directly involves the engaged receptor complexes. TCR/CD3 recruitment is accompanied by the accumulation of a series of prominent tyrosine-phosphorylated substrates and by an increase of the Lck activity in DIMs. Upon TCR stimulation, the DIM-associated receptor complexes are highly enriched in the hyperphosphorylated p23 zeta chains, contain most of the TCR/CD3 associated, phosphorylation-activated ZAP-70 kinases and seem to integrate into higher order, multiple tyrosine-phosphorylated substrate-containing protein complexes. The TCR/CD3 recruitment was found to depend on the activity of Src family kinases. We thus provide the first demonstration of recuitment of TCR/CD3 to DIMs upon receptor stimulation and propose it as a mechanism whereby TCR engagement is coupled to downstream signaling cascades. PMID- 9736614 TI - The Cdc42p effector Gic2p is targeted for ubiquitin-dependent degradation by the SCFGrr1 complex. AB - Cdc42p, a Rho-related GTP-binding protein, regulates cytoskeletal polarization and rearrangements in eukaryotic cells. In yeast, Gic1p and Gic2p are effectors of Cdc42p involved in actin polarization at bud emergence. Gic2p is expressed in a cell cycle-dependent manner and rapidly disappears shortly after bud emergence concomitant with the activation of the G1 cyclin-dependent kinase Cdc28p-Clnp. Here we have shown that the rapid disappearance of Gic2p results from ubiquitin dependent proteolysis. Biochemical and genetic evidence demonstrates that degradation of Gic2p required the Skp1-cullin-F-box protein complex (SCF) components Cdc34p, Cdc53p, Skp1p and Grr1p, but not Cdc4p. Phosphorylation of several C-terminal sites of Gic2p served as part of the recognition signal for ubiquitination. In addition, binding of Gic2p to Cdc42p was a prerequisite for degradation, suggesting that specifically the active form of Gic2p is targeted for destruction. Finally, our data indicate that degradation of Gic2p may be part of a mechanism which restricts cytoskeletal polarization in the G1 phase of the cell cycle. PMID- 9736615 TI - Identification and analysis of PH domain-containing targets of phosphatidylinositol 3-kinase using a novel in vivo assay in yeast. AB - Phosphatidylinositol 3-kinase (PI3K) mediates a variety of cellular responses by generating PtdIns(3,4)P2 and PtdIns(3,4,5)P3. These 3-phosphoinositides then function directly as second messengers to activate downstream signaling molecules by binding pleckstrin homology (PH) domains in these signaling molecules. We have established a novel assay in the yeast Saccharomyces cerevisiae to identify proteins that bind PtdIns(3,4)P2 and PtdIns(3,4,5)P3 in vivo which we have called TOPIS (Targets of PI3K Identification System). The assay uses a plasma membrane targeted Ras to complement a temperature-sensitive CDC25 Ras exchange factor in yeast. Coexpression of PI3K and a fusion protein of activated Ras joined to a PH domain known to bind PtdIns(3,4)P2 (AKT) or PtdIns(3,4,5)P3 (BTK) rescues yeast growth at the non-permissive temperature of 37 degreesC. Using this assay, we have identified several amino acids in the beta1-beta2 region of PH domains that are critical for high affinity binding to PtdIns(3,4)P2 and/or PtdIns(3,4,5)P3, and we have proposed a structural model for how these PH domains might bind PI3K products with high affinity. From these data, we derived a consensus sequence which predicts high-affinity binding to PtdIns(3, 4)P2 and/or PtdIns(3,4,5)P3, and we have identified several new PH domain-containing proteins that bind PI3K products, including Gab1, Dos, myosinX, and Sbf1. Use of this assay to screen for novel cDNAs which rescue yeast at the non-permissive temperature should provide a powerful approach for uncovering additional targets of PI3K. PMID- 9736616 TI - Apc10 and Ste9/Srw1, two regulators of the APC-cyclosome, as well as the CDK inhibitor Rum1 are required for G1 cell-cycle arrest in fission yeast. AB - Many eukaryotic cells arrest the cell cycle at G1 phase upon nutrient deprivation. In fission yeast, during nitrogen starvation, cells divide twice and arrest at G1. We have isolated a novel type of sterile mutant, which undergoes one additional S phase upon starvation and, as a result, arrests at G2. Three loci (apc10, ste9/srw1 and rum1) were identified. The apc10 mutants, previously unidentified, show, in addition to sterility, temperature-sensitive growth with defects in chromosome segregation. apc10(+) is essential for viability, encodes a conserved protein (a homologue of budding yeast Apc10/Doc1) and is required for ubiquitination and degradation of mitotic B-type cyclins. Apc10 does not co sediment with the 20S APC-cyclosome, a ubiquitin ligase for B-type cyclins, and in the apc10 mutant the 20S complex is intact, suggesting that it is a novel regulator for this complex. A subpopulation of Apc10 does co-immunoprecipitate with the anaphase-promoting complex (APC). A second gene, ste9(+)/srw1(+), encodes a member of the fizzy-related family, also regulators of the APC. Finally, Rum1 is a cyclin-dependent kinase (CDK) inhibitor which exists only in G1. The results suggest that dual downregulation of CDK, one via the APC and the other via the CDK inhibitor, is a universal mechanism that is used to arrest cell cycle progression at G1. PMID- 9736617 TI - Identification of a copper-induced intramolecular interaction in the transcription factor Mac1 from Saccharomyces cerevisiae. AB - Mac1 mediates copper (Cu)-dependent expression of genes involved in high-affinity uptake of copper ions in Saccharomyces cerevisiae. Mac1 is a transcriptional activator in Cu-deficient cells, but is inhibited in Cu-replete cells. Mac1 resides within the nucleus in both Cu-deficient and Cu-loaded cells. Cu inhibition of Mac1 appears to result from binding of eight copper ions within a C terminal segment consisting of two Cys-rich motifs. In addition, two zinc ions are bound within the N-terminal DNA-binding domain. Only 4-5 mol. eq. Cu are bound to a mutant Mac1 (His279Gln substitution) that is impervious to Cu inhibition. The CuMac1 complex is luminescent, indicative of copper bound in the Cu(I) state. Cu binding induces a molecular switch resulting in an intramolecular interaction in Mac1 between the N-terminal DNA-binding domain and the C-terminal activation domain. This allosteric interaction is Cu dependent and is not observed when Mac1 contained the mutant His279Gln substitution. Fusion of the minimal DNA-binding domain of Mac1 (residues 1-159) to the minimal Cu-binding activation domain (residues 252-341) yields a functional Cu-regulated transcriptional activator. These results suggest that Cu repression of Mac1 arises from a Cu-induced intramolecular interaction that inhibits both DNA binding and transactivation activities. PMID- 9736618 TI - Complex formation by the Drosophila MSL proteins: role of the MSL2 RING finger in protein complex assembly. AB - Drosophila MSL proteins are thought to act within a complex to elevate transcription from the male X chromosome. We found that the MSL1, MSL2 and MSL3 proteins are associated in immunoprecipitations, chromatographic steps and in the yeast two-hybrid system, but that the MLE protein is not tightly complexed in these assays. We focused our analysis on the MSL2-MSL1 interaction, which is postulated to play a critical role in MSL complex association with the X chromosome. Using a modified two-hybrid assay, we isolated missense mutations in MSL2 that disrupt its interaction with MSL1. Eleven out of 12 mutated residues clustered around the first zinc-binding site of the RING finger domain were conserved in a Drosophila virilis MSL2 homolog. Two pre-existing msl2 alleles, which fail to support male viability in vivo, have lesions in the same region of the RING finger. We tested these in the two-hybrid system and found that they are also defective in interaction with MSL1. Mutation of the second zinc-binding site had little effect on MSL1 binding, suggesting that this portion of the RING finger may have a distinct function. Our data support a model in which MSL2-MSL1 interaction nucleates assembly of an MSL complex, with which MLE is weakly or transiently associated. PMID- 9736619 TI - Antigenic variation in malaria: in situ switching, relaxed and mutually exclusive transcription of var genes during intra-erythrocytic development in Plasmodium falciparum. AB - Members of the Plasmodium falciparum var gene family encode clonally variant adhesins, which play an important role in the pathogenicity of tropical malaria. Here we employ a selective panning protocol to generate isogenic P.falciparum populations with defined adhesive phenotypes for CD36, ICAM-1 and CSA, expressing single and distinct var gene variants. This technique has established the framework for examining var gene expression, its regulation and switching. It was found that var gene switching occurs in situ. Ubiquitous transcription of all var gene variants appears to occur in early ring stages. However, var gene expression is tightly regulated in trophozoites and is exerted through a silencing mechanism. Transcriptional control is mutually exclusive in parasites that express defined adhesive phenotypes. In situ var gene switching is apparently mediated at the level of transcriptional initiation, as demonstrated by nuclear run-on analyses. Our results suggest that an epigenetic mechanism(s) is involved in var gene regulation. PMID- 9736620 TI - The deadenylating nuclease (DAN) is involved in poly(A) tail removal during the meiotic maturation of Xenopus oocytes. AB - Exonucleolytic degradation of the poly(A) tail is often the first step in the decay of eukaryotic mRNAs and is also used to silence certain maternal mRNAs translationally during oocyte maturation and early embryonic development. We previously described the purification of a poly(A)-specific 3'-exoribonuclease (deadenylating nuclease, DAN) from mammalian tissue. Here, the isolation and functional characterization of cDNA clones encoding human DAN is reported. Recombinant DAN overexpressed in Escherichia coli has properties similar to those of the authentic protein. The amino acid sequence of DAN shows homology to the RNase D family of 3'-exonucleases. DAN appears to be localized in both the nucleus and the cytoplasm. It is not stably associated with polysomes or ribosomal subunits. Xenopus oocytes contain nuclear and cytoplasmic DAN isoforms, both of which are closely related to the human DAN. Anti-DAN antibody microinjected into oocytes inhibits default deadenylation during progesterone induced maturation. Ectopic expression of human DAN in enucleated oocytes rescues maturation-specific deadenylation, indicating that amphibian and mammalian DANs are functionally equivalent. PMID- 9736621 TI - L-arginine recognition by yeast arginyl-tRNA synthetase. AB - The crystal structure of arginyl-tRNA synthetase (ArgRS) from Saccharomyces cerevisiae, a class I aminoacyl-tRNA synthetase (aaRS), with L-arginine bound to the active site has been solved at 2.75 A resolution and refined to a crystallographic R-factor of 19.7%. ArgRS is composed predominantly of alpha helices and can be divided into five domains, including the class I-specific active site. The N-terminal domain shows striking similarity to some completely unrelated proteins and defines a module which should participate in specific tRNA recognition. The C-terminal domain, which is the putative anticodon-binding module, displays an all-alpha-helix fold highly similar to that of Escherichia coli methionyl-tRNA synthetase. While ArgRS requires tRNAArg for the first step of the aminoacylation reaction, the results show that its presence is not a prerequisite for L-arginine binding. All H-bond-forming capability of L-arginine is used by the protein for the specific recognition. The guanidinium group forms two salt bridge interactions with two acidic residues, and one H-bond with a tyrosine residue; these three residues are strictly conserved in all ArgRS sequences. This tyrosine is also conserved in other class I aaRS active sites but plays several functional roles. The ArgRS structure allows the definition of a new framework for sequence alignments and subclass definition in class I aaRSs. PMID- 9736622 TI - Functional analysis of peptide motif for RNA microhelix binding suggests new family of RNA-binding domains. AB - RNA microhelices that recreate the acceptor stems of transfer RNAs are charged with specific amino acids. Here we identify a two-helix pair in alanyl-tRNA synthetase that is required for RNA microhelix binding. A single point mutation at an absolutely conserved residue in this motif selectively disrupts RNA binding without perturbation of the catalytic site. These results, and findings of similar motifs in the proximity of the active sites of other tRNA synthetases, suggest that two-helix pairs are widespread and provide a structural framework important for contacts with bound RNA substrates. PMID- 9736623 TI - Structure of the double-stranded RNA-binding domain of the protein kinase PKR reveals the molecular basis of its dsRNA-mediated activation. AB - Protein kinase PKR is an interferon-induced enzyme that plays a key role in the control of viral infections and cellular homeostasis. Compared with other known kinases, PKR is activated by a distinct mechanism that involves double-stranded RNA (dsRNA) binding in its N-terminal region in an RNA sequence-independent fashion. We report here the solution structure of the 20 kDa dsRNA-binding domain (dsRBD) of human PKR, which provides the first three-dimensional insight into the mechanism of its dsRNA-mediated activation. The structure of dsRBD exhibits a dumb-bell shape comprising two tandem linked dsRNA-binding motifs (dsRBMs) both with an alpha-beta-beta-beta-alpha fold. The structure, combined with previous mutational and biochemical data, reveals a highly conserved RNA-binding site on each dsRBM and suggests a novel mode of protein-RNA recognition. The central linker is highly flexible, which may enable the two dsRBMs to wrap around the RNA duplex for cooperative and high-affinity binding, leading to the overall change of PKR conformation and its activation. PMID- 9736624 TI - Crystal structure of restriction endonuclease BglI bound to its interrupted DNA recognition sequence. AB - The crystal structure of the type II restriction endonuclease BglI bound to DNA containing its specific recognition sequence has been determined at 2.2 A resolution. This is the first structure of a restriction endonuclease that recognizes and cleaves an interrupted DNA sequence, producing 3' overhanging ends. BglI is a homodimer that binds its specific DNA sequence with the minor groove facing the protein. Parts of the enzyme reach into both the major and minor grooves to contact the edges of the bases within the recognition half sites. The arrangement of active site residues is strikingly similar to other restriction endonucleases, but the co-ordination of two calcium ions at the active site gives new insight into the catalytic mechanism. Surprisingly, the core of a BglI subunit displays a striking similarity to subunits of EcoRV and PvuII, but the dimer structure is dramatically different. The BglI-DNA complex demonstrates, for the first time, that a conserved subunit fold can dimerize in more than one way, resulting in different DNA cleavage patterns. PMID- 9736625 TI - McrBs, a modulator peptide for McrBC activity. AB - McrBC is a methylation-dependent endonuclease from Escherichia coli K-12. The enzyme recognizes DNA with modified cytosines preceded by a purine. McrBC restricts DNA that contains at least two methylated recognition sites separated by 40-80 bp. Two gene products, McrBL and McrBs, are produced from the mcrB gene and one, McrC, from the mcrC gene. DNA cleavage in vitro requires McrBL, McrC, GTP and Mg2+. We found that DNA cleavage was optimal at a ratio of 3-5 McrBL per molecule of McrC, suggesting that formation of a multisubunit complex with several molecules of McrBL is required for cleavage. To understand the role of McrBs, we have purified the protein and analyzed its role in vitro. At the optimal ratio of 3-5 McrBL per molecule of McrC, McrBs acted as an inhibitor of DNA cleavage. Inhibition was due to sequestration of McrC and required the presence of GTP, suggesting that the interaction is GTP dependent. If McrC was in excess, a condition resulting in suboptimal DNA cleavage, addition of McrBs enhanced DNA cleavage, presumably due to sequestration of excess McrC. We suggest that the role of McrBs is to modulate McrBC activity by binding to McrC. PMID- 9736626 TI - A novel mode of DNA recognition by a beta-sheet revealed by the solution structure of the GCC-box binding domain in complex with DNA. AB - The 3D solution structure of the GCC-box binding domain of a protein from Arabidopsis thaliana in complex with its target DNA fragment has been determined by heteronuclear multidimensional NMR in combination with simulated annealing and restrained molecular dynamic calculation. The domain consists of a three-stranded anti-parallel beta-sheet and an alpha-helix packed approximately parallel to the beta-sheet. Arginine and tryptophan residues in the beta-sheet are identified to contact eight of the nine consecutive base pairs in the major groove, and at the same time bind to the sugar phosphate backbones. The target DNA bends slightly at the central CG step, thereby allowing the DNA to follow the curvature of the beta sheet. PMID- 9736627 TI - Homologous recombination and non-homologous end-joining pathways of DNA double strand break repair have overlapping roles in the maintenance of chromosomal integrity in vertebrate cells. AB - Eukaryotic cells repair DNA double-strand breaks (DSBs) by at least two pathways, homologous recombination (HR) and non-homologous end-joining (NHEJ). Rad54 participates in the first recombinational repair pathway while Ku proteins are involved in NHEJ. To investigate the distinctive as well as redundant roles of these two repair pathways, we analyzed the mutants RAD54(-/-), KU70(-/-) and RAD54(-/-)/KU70(-/-), generated from the chicken B-cell line DT40. We found that the NHEJ pathway plays a dominant role in repairing gamma-radiation-induced DSBs during G1-early S phase while recombinational repair is preferentially used in late S-G2 phase. RAD54(-/-)/KU70(-/-) cells were profoundly more sensitive to gamma-rays than either single mutant, indicating that the two repair pathways are complementary. Spontaneous chromosomal aberrations and cell death were observed in both RAD54(-/-) and RAD54(-/-)/KU70(-/-) cells, with RAD54(-/-)/KU70(-/-) cells exhibiting significantly higher levels of chromosomal aberrations than RAD54(-/-) cells. These observations provide the first genetic evidence that both repair pathways play a role in maintaining chromosomal DNA during the cell cycle. PMID- 9736628 TI - An ATP-ADP switch in MuB controls progression of the Mu transposition pathway. AB - MuB protein, an ATP-dependent DNA-binding protein, collaborates with Mu transposase to promote efficient transposition. MuB binds target DNA, delivers this target DNA segment to transposase and activates transposase's catalytic functions. Using ATP-bound, ADP-bound and ATPase-defective MuB proteins we investigated how nucleotide binding and hydrolysis control the activities of MuB protein, important for transposition. We found that both MuB-ADP and MuB-ATP stimulate transposase, whereas only MuB-ATP binds with high affinity to DNA. Four different ATPase-defective MuB mutants fail to activate the normal transposition pathway, further indicating that ATP plays critical regulatory roles during transposition. These mutant proteins fall into two classes: class I mutants are defective in target DNA binding, whereas class II mutants bind target DNA, deliver it to transposase, but fail to promote recombination with this DNA. Based on these studies, we propose that the switch from the ATP- to ADP-bound form allows MuB to release the target DNA while maintaining its stimulatory interaction with transposase. Thus, ATP-hydrolysis by MuB appears to function as a molecular switch controlling how target DNA is delivered to the core transposition machinery. PMID- 9736629 TI - Identification of a survival-promoting peptide in medium conditioned by oxidatively stressed cell lines of nervous system origin. AB - A survival-promoting peptide has been purified from medium conditioned by Y79 human retinoblastoma cells and a mouse hippocampal cell line (HN 33.1) exposed to H2O2. A 30 residue synthetic peptide was made on the basis of N-terminal sequences obtained during purification, and it was found to exhibit gel mobility and staining properties similar to the purified molecules. The peptide maintains cells and their processes in vitro for the HN 33.1 cell line treated with H2O2, and in vivo for cortical neurons after lesions of the cerebral cortex. It has weak homology with a fragment of a putative bacterial antigen and, like that molecule, binds IgG. The peptide also contains a motif reminiscent of a critical sequence in the catalytic region of calcineurin-type phosphatases; surprisingly, like several members of this family, the peptide catalyzes the hydrolysis of para nitrophenylphosphate in the presence of Mn2+. Application of the peptide to one side of bilateral cerebral cortex lesions centered on area 2 in rats results in an increase in IgG immunoreactivity in the vicinity of the lesions 7 d after surgery. Microglia immunopositive for IgG and ED-1 are, however, dramatically reduced around the lesions in the treated hemisphere. Furthermore, pyramidal neurons that would normally shrink, die, or disintegrate were maintained, as determined by MAP2 immunocytochemistry and Nissl staining. These survival effects were often found in both hemispheres. The results suggest that this peptide operates by diffusion to regulate the immune response and thereby rescue neurons that would usually degenerate after cortical lesions. The phosphatase activity of this molecule also suggests the potential for direct neuron survival-promoting effects. PMID- 9736630 TI - Tau cleavage and dephosphorylation in cerebellar granule neurons undergoing apoptosis. AB - Cerebellar granule cells undergo apoptosis in culture after deprivation of potassium and serum. During this process we found that tau, a neuronal microtubule-associated protein that plays a key role in the maintenance of neuronal architecture, and the pathology of which correlates with intellectual decline in Alzheimer's disease, is cleaved. The final product of this cleavage is a soluble dephosphorylated tau fragment of 17 kDa that is unable to associate with microtubules and accumulates in the perikarya of dying cells. The appearance of this 17 kDa fragment is inhibited by both caspase and calpain inhibitors, suggesting that tau is an in vivo substrate for both of these proteases during apoptosis. Tau cleavage is correlated with disruption of the microtubule network, and experiments with colchicine and taxol show that this is likely to be a cause and not a consequence of tau cleavage. These data indicate that tau cleavage and change in phosphorylation are important early factors in the failure of the microtubule network that occurs during neuronal apoptosis. Furthermore, this study introduces new insights into the mechanism(s) that generate the truncated forms of tau present in Alzheimer's disease. PMID- 9736631 TI - Nicotinic stimulation produces multiple forms of increased glutamatergic synaptic transmission. AB - Synaptic modulation and long-term synaptic changes are thought to be the cellular correlates for learning and memory (Madison et al., 1991; Aiba et al., 1994, Goda and Stevens, 1996). The hippocampus is a center for learning and memory that receives abundant cholinergic innervation and has a high density of nicotinic acetylcholine receptors (nAChRs) (Wada et al., 1989; Woolf, 1991). We report that stro ng, brief stimulation of nAChRs enhanced hippocampal glutamatergic synaptic transmission on two independent time scales and altered the relationship between consecutively evoked synaptic currents. The nicotinic synaptic enhancement required extracellular calcium and was produced by the activation of presynaptic alpha7-containing nAChRs. Although one form of glutamatergic enhancement lasted only for seconds, another form lasted for minutes after the nicotinic stimulation had ceased and the nicotinic agonist had been washed away. The synaptic enhancement lasting minutes suggests that nAChR activity can initiate calcium dependent mechanisms that are known to induce glutamatergic synaptic plasticity. The results with evoked synaptic currents showed that nAChR activity can alter the relationship between the incoming presynaptic activity and outgoing postsynaptic signaling along glutamatergic fibers. Thus, the same information arriving along the same glutamatergic afferents will be processed differently when properly timed nicotinic activity converges onto the glutamatergic presynaptic terminals. Influencing information processing at glutamatergic synapses may be one way in which nicotinic cholinergic activity influences cognitive processes. Disruption of these nicotinic cholinergic mechanisms may contribute to the deficits associated with the degeneration of cholinergic functions during Alzheimer's disease. PMID- 9736632 TI - Opioid inhibition of hippocampal interneurons via modulation of potassium and hyperpolarization-activated cation (Ih) currents. AB - The actions of mu- and delta-opioid agonists (DAMGO and DPDPE, respectively) on GABAergic interneurons in stratum oriens of area CA1 of the hippocampus were examined by using whole-cell voltage-clamp recordings in brain slices. Both agonists consistently generated outward currents of similar magnitude (15-20 pA) in the majority of cells. However, under control conditions, current-voltage (I/V) relationships revealed that only a small number of these cells (3 of 77) demonstrated clear increases in membrane conductance, associated with the activation of the potassium current known as Girk. These interneurons also exhibited a slowly activating, inwardly rectifying current known as Ih on hyperpolarizing step commands. Ih was blocked by the extracellular application of cesium (3-9 mM) or ZD 7288 (10-100 microM) but was insensitive to barium (1-2 mM). In an effort to determine whether the holding current changes were attributable to the modulation of Girk and/or Ih, we used known blockers of these ion channels (barium or cesium and ZD 7288, respectively). Extracellular application of cesium (3-9 mM) or ZD 7288 (25-100 microM) blocked Ih and significantly reduced the opioid-induced outward currents by 58%. Under these conditions the opioid agonists activated a potassium current with characteristics similar to Girk. Similarly, during barium (1-2 mM) application the opioid-induced outward currents were reduced by 46%, and a clear reduction in Ih and the whole cell conductance was revealed. These data suggest that the opioids can modulate both Ih and Girk in the same population of stratum oriens interneurons and that the modulation of these ion channels can contribute to the inhibition of interneuron activity in the hippocampus. PMID- 9736633 TI - Anion currents and predicted glutamate flux through a neuronal glutamate transporter. AB - Kinetic properties of a native, neuronal glutamate transporter were studied by using rapid applications of glutamate to outside-out patches excised from Purkinje neurons. Pulses of glutamate activated anion currents associated with the transporter that were weakly antagonized by the transporter antagonist kainate. In addition, kainate blocked a resting anion conductance observed in the absence of glutamate. Transporter currents in response to glutamate concentration jumps under a variety of conditions were used to construct a cyclic kinetic model of the transporter. The model simulates both the anion conductance and the glutamate flux through the transporter, thereby permitting several predictions regarding the dynamics of glutamate transport at the synapse. For example, the concentration-dependent binding rate of glutamate to the transporter is high, similar to binding rates suggested for ligand-gated glutamate receptors. At saturating glutamate concentrations, transporters cycle at a steady-state rate of 13/sec. Transporters are predicted to have a high efficiency; once bound, a glutamate molecule is more likely to be transported than to unbind. Physiological concentrations of internal sodium and glutamate significantly slow net transport. Finally, a fixed proportion of anion and glutamate flux is expected over a wide range of circumstances, providing theoretical support for using net charge flux to estimate the amount and time course of glutamate transport. PMID- 9736634 TI - Dopamine inactivates tryptophan hydroxylase and forms a redox-cycling quinoprotein: possible endogenous toxin to serotonin neurons. AB - Exposure of tryptophan hydroxylase (TPH), the initial and rate-limiting enzyme in the biosynthesis of the neurotransmitter serotonin, to dopamine under mild oxidizing conditions (iron + H2O2) or in the presence of tyrosinase results in a concentration-dependent inactivation of the enzyme. Dopamine, iron, H2O2, or tyrosinase alone does not alter TPH activity. Similarly, N-acetyldopamine oxidized with one equivalent of sodium periodate causes a concentration-dependent inactivation of TPH as well. TPH is protected from dopamine-induced inactivation by reduced glutathione, ascorbic acid, and dithiothreitol but not by the radical scavengers DMSO, mannitol, or superoxide dismutase. Parallel studies with [3H]dopamine reveal a high negative correlation between inhibition of catalysis and incorporation of tritium into the enzyme. Those reducing agents and antioxidants that protect TPH from inactivation are effective in preventing the labeling of TPH by [3H]dopamine. Acid hydrolysis and HPLC with electrochemical detection (HPLC-EC) analysis of inactivated TPH revealed the formation of cysteinyl-dopamine residues within the enzyme. Exposure of dopamine-modified TPH to redox-cycling staining after SDS-PAGE confirmed the formation of a quinoprotein. These results indicate that dopamine-quinones covalently modify cysteinyl residues in TPH, leading directly to the loss of catalytic activity, and establish that TPH could be a target for dopamine-quinones in vivo after drugs (e.g., neurotoxic amphetamines) that cause dopamine-dependent inactivation of TPH. Redox cycling of a TPH-quinoprotein could also participate in the serotonin neuronal toxicity caused by these same drugs. PMID- 9736635 TI - Postsynaptic complex spike bursting enables the induction of LTP by theta frequency synaptic stimulation. AB - Long-term potentiation (LTP), a persistent enhancement of synaptic transmission that may be involved in some forms of learning and memory, is induced at excitatory synapses in the CA1 region of the hippocampus by coincident presynaptic and postsynaptic activity. Although action potentials back propagating into dendrites of hippocampal pyramidal cells provide sufficient postsynaptic activity to induce LTP under some in vitro conditions, it is not known whether LTP can be induced by patterns of postsynaptic action potential firing that occur in these cells in vivo. Here we report that a characteristic in vivo pattern of action potential generation in CA1 pyramidal cells known as the complex spike burst enables the induction of LTP during theta frequency synaptic stimulation in the CA1 region of hippocampal slices maintained in vitro. Our results suggest that complex spike bursting may have an important role in synaptic processes involved in learning and memory formation, perhaps by producing a highly sensitive postsynaptic state during which even low frequencies of presynaptic activity can induce LTP. PMID- 9736636 TI - Involvement of stretch-activated Cl- channels in ramification of murine microglia. AB - A stretch-activated Cl- current (ICl) was investigated in cultured murine microglia using the whole-cell configuration of the patch-clamp technique. After application of membrane stretch, a Cl- current appeared within seconds, and its amplitude increased further within 3-8 min. ICl underwent rundown, which was prevented by addition of 4 mM ATP to the intracellular perfusing solution. The stretch-activated Cl- current exhibited outward rectification and did not show any voltage-dependent gating. Lowering the concentration of extracellular Cl- from 142 to 12 mM by equimolar substitution of Cl- with gluconate shifted the reversal potential of ICl by 41.6 +/- 1.8 mV in the depolarizing direction. 4, 4' Diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS) and 4-acetamido-4' isothiocyanatostilbene-2,2'-disulfonic acid (SITS) blocked ICl in a voltage- and time-dependent manner. At a test potential of +40 mV, a half-maximal blockade at 16.1 microM DIDS and at 71.0 microM SITS was determined for ICl. At a concentration of 200 microM, 5-nitro-2-(3-phenylpropylamino)benzoic acid or flufenamic acid blocked ICl by 88% and 75%, respectively. Each of these four Cl- channel blockers reversibly inhibited the ramification process of microglia, whereas blockers of voltage-gated Na+ and K+ channels did not affect the transformation of microglia from their ameboid into the ramified phenotype. It is suggested that in microglia functional stretch-activated Cl- channels are required for the induction of ramification but not for maintaining the ramified shape. PMID- 9736637 TI - Functional redundancy of FMRFamide-related peptides at the Drosophila larval neuromuscular junction. AB - The Drosophila FMRFamide gene encodes multiple FMRFamide-related peptides. These peptides are expressed by neurosecretory cells and may be released into the blood to act as neurohormones. We analyzed the effects of eight of these peptides on nerve-stimulated contraction (twitch tension) of Drosophila larval body-wall muscles. Seven of the peptides strongly enhanced twitch tension, and one of the peptides was inactive. Their targets were distributed widely throughout the somatic musculature. The effects of one peptide, DPKQDFMRFamide, were unchanged after the onset of metamorphosis. The seven active peptides showed similar dose response curves. Each had a threshold concentration near 1 nM, and the EC50 for each peptide was approximately 40 nM. At concentrations <0.1 microM, the responses to each of the seven excitatory peptides followed a time course that matched the fluctuations of the peptide concentration in the bath. At higher concentrations, twitch tension remained elevated for 5-10 min or more after wash out of the peptide. When the peptides were presented as mixtures predicted by their stoichiometric ratios in the dFMRFamide propeptide, the effects were additive, and there were no detectable higher-order interactions among them. One peptide was tested and found to enhance synaptic transmission. At 0.1 microM, DPKQDFMRFamide increased the amplitude of the excitatory junctional current to 151% of baseline within 3 min. Together, these results indicate that the products of the Drosophila FMRFamide gene function as neurohormones to modulate the strength of contraction at the larval neuromuscular junction. In this role these seven peptides appear to be functionally redundant. PMID- 9736638 TI - Central P2X4 and P2X6 channel subunits coassemble into a novel heteromeric ATP receptor. AB - Ionotropic ATP receptors are widely expressed in mammalian CNS. Despite extensive functional characterization of neuronal homomeric P2X receptors in heterologous expression systems, the subunit composition of native central P2X ATP-gated channels remains to be elucidated. P2X4 and P2X6 are major central subunits with highly overlapping mRNA distribution at both regional and cellular levels. When expressed alone in Xenopus oocytes, P2X6 subunits do not assemble into surface receptors responsive to ATP applications. On the other hand, P2X4 subunits assemble into bona fide ATP-gated channels, slowly desensitizing and weakly sensitive to the partial agonist alpha,beta-methylene ATP and to noncompetitive antagonists suramin and pyridoxal-5-phosphate-6-azophenyl-2',4'-disulfonic acid. We demonstrate here that the coexpression of P2X4 and P2X6 subunits in Xenopus oocytes leads to the generation of a novel pharmacological phenotype of ionotropic ATP receptors. Heteromeric P2X4+6 receptors are activated by low micromolar alpha, beta-methylene ATP (EC50 = 12 microM) and are blocked by suramin and by Reactive Blue 2, which has the property, at low concentrations, to potentiate homomeric P2X4 receptors. The assembly of P2X4 with P2X6 subunits results from subunit-dependent interactions, as shown by their specific copurification from HEK-293 cells transiently transfected with various epitope tagged P2X channel subunits. Our data strongly suggest that the numerous cases of neuronal colocalizations of P2X4 and P2X6 subunits observed in mammalian CNS reflect the native expression of heteromeric P2X4+6 channels with unique functional properties. PMID- 9736639 TI - Selective histamine uptake rescues photo- and mechanoreceptor function of histidine decarboxylase-deficient Drosophila mutant. AB - In insects, histamine is found both in the peripheral nervous system (PNS) and in the CNS and is known to function as a fast neurotransmitter in photoreceptors that have been shown to express selectively the hdc gene. This gene codes for histidine decarboxylase (HDC), the enzyme for histamine synthesis. Fast neurotransmission requires the efficient removal of the transmitter from the synaptic cleft. Here we identify in Drosophila photo- and mechanoreceptors a histamine uptake mechanism that can restore the function of these receptors in mutants unable to synthesize histamine. When apparent null mutants for the hdc gene imbibe aqueous histamine solution or are genetically "rescued" by a transgene ubiquitously expressing histidine decarboxylase under heat-shock control, sufficient amounts of histamine selectively accumulate in photo- and mechanoreceptors to generate near-normal electrical responses in second-order visual interneurons and qualitatively to restore wild-type visual and mechanosensory behavior. This strongly supports the proposal that histamine functions as a fast neurotransmitter also in a certain class of mechanoreceptors. A set of CNS-intrinsic neurons that in the wild type contain high concentrations of histamine apparently lacks this uptake mechanism. We therefore speculate that histamine of intrinsic neurons may function as a neuromodulator rather than as a fast transmitter. PMID- 9736640 TI - Expression cloning and characterization of NSIST, a novel sulfotransferase expressed by a subset of neurons and postsynaptic targets. AB - Synapses are distinguished by localized concentrations of specific proteins, many of which bear the marks of posttranslational processing such as glycosylation and sulfation. One strategy to elucidate this posttranslational tailoring is to identify the enzymes that create these modifications. Monoclonal antibody 3B3 recognizes a carbohydrate-containing epitope expressed on dystroglycan and other constituents of Torpedo electric organ synaptic membranes. We used mAb 3B3 in an immunofluorescence-based expression-cloning method and isolated a cDNA clone conferring mAb-3B3 immunoreactivity to transfected COS cells. The deduced polypeptide has a predicted molecular weight of 51 kDa, a type II transmembrane topology, and four potential N-linked glycosylation sites. The polypeptide, which we term NSIST (nervous system involved sulfotransferase), shows extensive, although not complete, homology to a chondroitin-6-sulfotransferase and limited homology to other sulfotransferases. In NSIST-transfected COS cells, 35SO4 incorporation and chondroitin-sulfate-like immunoreactivity are increased. In vivo, NSIST occurs as a single 2.4 kb transcript abundant in Torpedo electric organ, moderately expressed in spinal cord and electric lobe, and undetectable in non-neural tissues. Immunohistochemistry shows that NSIST is expressed in a punctate distribution in the innervated portion of electrocytes. In the CNS, NSIST-like immunoreactivity is localized within the somas of motor neurons and neurons of the electromotor nucleus, whereas mAb-3B3 immunostaining is associated with cell surfaces and neuropil. Neuronal NSIST is therefore likely to exert its effects extracellularly; although NSIST is synthesized by neurons, its product, the 3B3 epitope, is found outside neuronal cell bodies. Our evidence indicates that NSIST participates in nervous system specific posttranslational modifications, perhaps including those at synapses. PMID- 9736642 TI - Imaging spreading depression and associated intracellular calcium waves in brain slices. AB - Spreading depression (SD) was analyzed in hippocampal and neocortical brain slices by imaging intrinsic optical signals in combination with either simultaneous electrophysiological recordings or imaging of intracellular calcium dynamics. The goal was to determine the roles of intracellular calcium (Ca2+int) waves in the generation and propagation of SD. Imaging of intrinsic optical signals in the hippocampus showed that ouabain consistently induced SD, which characteristically started in the CA1 region, propagated at 15-35 micrometer/sec, and traversed across the hippocampal fissure to the dentate gyrus. In the dendritic regions of both CA1 and the dentate gyrus, SD caused a transient increase in light transmittance, characterized by both a rapid onset and a rapid recovery. In contrast, in the cell body regions the transmittance increase was prolonged. Simultaneous imaging of intracellular calcium and intrinsic optical signals revealed that a slow Ca2+int increase preceded any change in transmittance. Additionally, a wave of increased Ca2+int typically propagated many seconds ahead of the change in transmittance. These calcium increases were also observed in individual astrocytes injected with calcium orange, indicating that Ca2+int waves were normally associated with SD. However, when hippocampal slices were incubated in calcium-free/EGTA external solutions, SD was still observed, although Ca2+int waves were completely abolished. Under these conditions SD had a comparable peak increase in transmittance but a slower onset and a faster recovery. These results demonstrate that although there are calcium dynamics associated with SD, these increases are not necessary for the initiation or propagation of spreading depression. PMID- 9736641 TI - Dynamic regulation of RGS2 suggests a novel mechanism in G-protein signaling and neuronal plasticity. AB - Long-term neuronal plasticity is known to be dependent on rapid de novo synthesis of mRNA and protein, and recent studies provide insight into the molecules involved in this response. Here, we demonstrate that mRNA encoding a member of the regulator of G-protein signaling (RGS) family, RGS2, is rapidly induced in neurons of the hippocampus, cortex, and striatum in response to stimuli that evoke plasticity. Although several members of the RGS family are expressed in brain with discrete neuronal localizations, RGS2 appears unique in that its expression is dynamically responsive to neuronal activity. In biochemical assays, RGS2 stimulates the GTPase activity of the alpha subunit of Gq and Gi1. The effect on Gi1 was observed only after reconstitution of the protein in phospholipid vesicles containing M2 muscarinic acetylcholine receptors. RGS2 also inhibits both Gq- and Gi-dependent responses in transfected cells. These studies suggest a novel mechanism linking neuronal activity and signal transduction. PMID- 9736643 TI - Temperature-sensitive neuromuscular transmission in Kv1.1 null mice: role of potassium channels under the myelin sheath in young nerves. AB - In mammalian myelinated nerves, the internodal axon that is normally concealed by the myelin sheath expresses a rich repertoire of K channel subtypes thought to be important in modulating action potential propagation. The function of myelin covered K channels at transition zones, however, has remained unexplored. Here we show that deleting the voltage-sensitive potassium channel Kv1.1 from mice confers a marked temperature-sensitivity to neuromuscular transmission in postnatal day 14 (P14)-P21 mice. Using immunofluorescence and electrophysiology, we examined contributions of four regions of the peripheral nervous system to the mutant phenotype: the nerve trunk, the myelinated segment preceding the terminal, the presynaptic terminal membrane itself, and the muscle. We conclude that the temperature-sensitive neuromuscular transmission is accounted for solely by a deficiency in Kv1.1 normally concealed in the myelinated segments just preceding the terminal. This paper demonstrates that under certain situations of physiological stress, the functional role of myelin-covered K channels is dramatically enhanced as the transition zone at the neuromuscular junction is approached. PMID- 9736644 TI - Altered Ca2+ signaling and mitochondrial deficiencies in hippocampal neurons of trisomy 16 mice: a model of Down's syndrome. AB - It has been suggested that augmented nerve cell death in neurodegenerative diseases might result from an impairment of mitochondrial function. To test this hypothesis, we investigated age-dependent changes in neuronal survival and glutamate effects on Ca2+ homeostasis and mitochondrial energy metabolism in cultured hippocampal neurons from diploid and trisomy 16 (Ts16) mice, a model of Down's syndrome. Microfluorometric techniques were used to measure survival rate, [Ca2+]i level, mitochondrial membrane potential, and NAD(P)H autofluorescence. We found that Ts16 neurons die more than twice as fast as diploid neurons under otherwise identical culture conditions. Basal [Ca2+]i levels were elevated in Ts16 neurons. Moreover, in comparison to diploid neurons, Ts16 neurons showed a prolonged recovery of [Ca2+]i and mitochondrial membrane potential after brief glutamate application. Glutamate evoked an initial NAD(P)H decrease that was found to be extended in Ts16 neurons in comparison to diploid neurons. Furthermore, for all age groups tested, glutamate failed to cause a subsequent NAD(P)H overshoot in Ts16 cultures in contrast to diploid cultures. In the presence of cyclosporin A, an inhibitor of the mitochondrial membrane permeability transition, NAD(P)H increase was observed in both diploid and Ts16 neurons. The results support the hypothesis that Ca2+ impairs mitochondrial energy metabolism and may play a role in the pathogenesis of neurodegenerative changes in neurons from Ts16 mice. PMID- 9736645 TI - Intracellular calcium and cell death during ischemia in neonatal rat white matter astrocytes in situ. AB - The major pathological correlate of cerebral palsy is ischemic injury of CNS white matter. Histological studies show early injury of glial cells and axons. To investigate glial cell injury, I monitored intracellular Ca2+ and cell viability in fura-2-loaded neonatal rat white matter glial cells during ischemia. Fura-2 fixation combined with immunohistochemistry revealed that fura-2-loaded cells were GFAP+/O4(-) and were therefore a population of neonatal white matter astrocytes. Significant ischemic Ca2+ influx was found, mediated by both L- and T type voltage-gated Ca2+ channels. Ca2+ influx via T-type channels was the most important factor during the initial stage of ischemia and was associated with significant cell death within 10-20 min of the onset of ischemia. The Na+-Ca2+ exchanger acted to remove cytoplasmic Ca2+ throughout the ischemic and recovery periods. Neither the release of Ca2+ from intracellular stores nor influx via glutamate-gated channels contributed to the rise in intracellular Ca2+ during ischemia. Ischemic cell death was reduced significantly by removing extracellular Ca2+ or by blocking voltage-gated Ca2+ channels. The exclusively voltage-gated Ca2+ channel nature of the Ca2+ influx, the role played by T-type Ca2+ channels, the protective effect of the Na+-Ca2+ exchanger, and the lack of significant Ca2+ release from intracellular stores are features of ischemia that have not been reported in other CNS cell types. PMID- 9736646 TI - The nitric oxide-cGMP pathway may mediate communication between sensory afferents and projection neurons in the antennal lobe of Manduca sexta. AB - The nitric oxide (NO)-cGMP signaling system is thought to play important roles in the function of the olfactory system in both vertebrates and invertebrates. One way of studying the role of NO in the nervous system is to study the distribution and properties of NO synthase (NOS), as well as the soluble guanylyl cyclases (sGCs), which are the best characterized targets of NO. We study NOS and sGC in the relatively simple and well characterized insect olfactory system of the hawkmoth, Manduca sexta. We have cloned Manduca sexta nitric oxide synthase (MsNOS) and two sGCs (MsGCalpha1 and MsGCbeta1), characterized their basic biochemical properties, and studied their expression in the olfactory system. The sequences of the Manduca genes are highly similar to their mammalian homologs and show similar biochemical properties when expressed in COS-7 cells. In particular, we find that MsGC functions as an obligate heterodimer that is stimulated significantly by NO. We also find that MsNOS has a Ca2+-sensitive NO-producing activity similar to that of mammalian neuronal NOS. Northern and in situ hybridization analyses show that MsNOS and the MsGCs are expressed in a complementary pattern, with MsNOS expressed at high levels in the antennae and the MsGCs expressed at high levels in a subset of antennal lobe neurons. The expression patterns of these genes suggest that the NO-sGC signaling system may play a role in mediating communication between olfactory receptor neurons and projection neurons in the glomeruli of the antennal lobe. PMID- 9736647 TI - Neurotrophins induce formation of functional excitatory and inhibitory synapses between cultured hippocampal neurons. AB - Cell cultures were used to analyze the role of brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3) in the development of synaptic transmission. Neurons obtained from embryonic day 18 (E18) rat hippocampus and cultured for 2 weeks exhibited extensive spontaneous synaptic activity. By comparison, neurons obtained from E16 hippocampus expressed very low levels of spontaneous or evoked synaptic activity. Neurotrophin treatment produced a sevenfold increase in the number of functional synaptic connections in the E16 cultures. BDNF induced formation of both excitatory and inhibitory synapses, whereas NT-3 induced formation of only excitatory synapses. These effects were independent of serum or the age of the glia bed used for the culture. They were not accompanied by significant changes in synaptic-vesicle-associated proteins or glutamate receptors. Treatment of the cultures with the neurotrophins for 3 d was sufficient to establish the maximal level of functional synapses. During this period, neurotrophins did not affect the viability or the morphology of the excitatory neurons, although they did produce an increase in the number and length of dendrites of the GABAergic neurons. Remarkably, only BDNF caused an increase in the number of axonal branches and in the total length of the axons of the GABAergic neurons. These results support a unique and differential role for neurotrophins in the formation of excitatory and inhibitory synapses in the developing hippocampus. PMID- 9736648 TI - TrkB and neurotrophin-4 are important for development and maintenance of sympathetic preganglionic neurons innervating the adrenal medulla. AB - The adrenal medulla receives its major presynaptic input from sympathetic preganglionic neurons that are located in the intermediolateral (IML) column of the thoracic spinal cord. The neurotrophic factor concept would predict that these IML neurons receive trophic support from chromaffin cells in the adrenal medulla. We show here that adrenal chromaffin cells in the adult rat store neurotrophin (NT)-4, but do not synthesize or store detectable levels of BDNF or NT-3, respectively. Preganglionic neurons to the adrenal medulla identified by retrograde tracing with fast blue or Fluoro-Gold (FG) express TrkB mRNA. After unilateral destruction of the adrenal medulla, 24% of IML neurons, i.e., all neurons that are preganglionic to the adrenal medulla in spinal cord segments T7 T10, disappear. Administration of NT-4 in gelfoams (6 microgram) implanted into the medullectomized adrenal gland rescued all preganglionic neurons as evidenced by their presence after 4 weeks. NT-3 and cytochrome C were not effective. The action of NT-4 is accompanied by massive sprouting of axons in the vicinity of the NT-4 source as monitored by staining for acetylcholinesterase and synaptophysin immunoreactivity, suggesting that NT-4 may enlarge the terminal field of preganglionic nerves and enhance their access to trophic factors. Analysis of TrkB-deficient mice revealed degenerative changes in axon terminals on chromaffin cells. Furthermore, numbers of FG-labeled IML neurons in spinal cord segments T7-T10 of NT-4-deficient adult mice were significantly reduced. These data are consistent with the notion that NT-4 from chromaffin cells operates through TrkB receptors to regulate development and maintenance of the preganglionic innervation of the adrenal medulla. PMID- 9736649 TI - Activity of the delta-opioid receptor is partially reduced, whereas activity of the kappa-receptor is maintained in mice lacking the mu-receptor. AB - Previous pharmacological studies have indicated the possible existence of functional interactions between mu-, delta- and kappa-opioid receptors in the CNS. We have investigated this issue using a genetic approach. Here we describe in vitro and in vivo functional activity of delta- and kappa-opioid receptors in mice lacking the mu-opioid receptor (MOR). Measurements of agonist-induced [35S]GTPgammaS binding and adenylyl cyclase inhibition showed that functional coupling of delta- and kappa-receptors to G-proteins is preserved in the brain of mutant mice. In the mouse vas deferens bioassay, deltorphin II and cyclic[D penicillamine2, D-penicillamine5] enkephalin exhibited similar potency to inhibit smooth muscle contraction in both wild-type and MOR -/- mice. delta-Analgesia induced by deltorphin II was slightly diminished in mutant mice, when the tail flick test was used. Deltorphin II strongly reduced the respiratory frequency in wild-type mice but not in MOR -/- mice. Analgesic and respiratory responses produced by the selective kappa-agonist U-50,488H were unchanged in MOR-deficient mice. In conclusion, the preservation of delta- and kappa-receptor signaling properties in mice lacking mu-receptors provides no evidence for opioid receptor cross-talk at the cellular level. Intact antinociceptive and respiratory responses to the kappa-agonist further suggest that the kappa-receptor mainly acts independently from the mu-receptor in vivo. Reduced delta-analgesia and the absence of delta-respiratory depression in MOR-deficient mice together indicate that functional interactions may take place between mu-receptors and central delta-receptors in specific neuronal pathways. PMID- 9736650 TI - Regional selective neuronal degeneration after protein phosphatase inhibition in hippocampal slice cultures: evidence for a MAP kinase-dependent mechanism. AB - The regional selectivity and mechanisms underlying the toxicity of the serine/threonine protein phosphatase inhibitor okadaic acid (OA) were investigated in hippocampal slice cultures. Image analysis of propidium iodide labeled cultures revealed that okadaic acid caused a dose- and time-dependent injury to hippocampal neurons. Pyramidal cells in the CA3 region and granule cells in the dentate gyrus were much more sensitive to okadaic acid than the pyramidal cells in the CA1 region. Electron microscopy revealed ultrastructural changes in the pyramidal cells that were not consistent with an apoptotic process. Treatment with okadaic acid led to a rapid and sustained tyrosine phosphorylation of the mitogen-activated protein kinases ERK1 and ERK2 (p44/42(mapk)). The phosphorylation was markedly reduced after treatment of the cultures with the microbial alkaloid K-252a (a nonselective protein kinase inhibitor) or the MAP kinase kinase (MEK1/2) inhibitor PD98059. K-252a and PD98059 also ameliorated the okadaic acid-induced cell death. Inhibitors of protein kinase C, Ca2+/calmodulin-dependent protein kinase II, or tyrosine kinase were ineffective. These results indicate that sustained activation of the MAP kinase pathway, as seen after e.g., ischemia, may selectively harm specific subsets of neurons. The susceptibility to MAP kinase activation of the CA3 pyramidal cells and dentate granule cells may provide insight into the observed relationship between cerebral ischemia and dementia in Alzheimer's disease. PMID- 9736651 TI - Perforin-dependent neurologic injury in a viral model of multiple sclerosis. AB - In this study we demonstrate perforin-mediated cytotoxic effector function is necessary for viral clearance and may directly contribute to the development of neurologic deficits after demyelination in the Theiler's murine encephalomyelitis virus (TMEV) model of multiple sclerosis. We previously demonstrated major histocompatability complex (MHC) class I-deficient (beta2m-deficient) mice with an otherwise resistant genotype develop severe demyelination with minimal neurologic disease when chronically infected with TMEV. These studies implicate CD8(+) T cells as the pathogenic cell in the induction of neurologic disease after demyelination. To determine which effector mechanisms of CD8(+) T cells, granule exocytosis or Fas ligand expression, play a role in the development of demyelination and clinical disease, we infected perforin-deficient, lpr (Fas mutation), and gld (Fas ligand mutation) mice with TMEV. Perforin-deficient mice showed viral persistence in the CNS, chronic brain pathology, and demyelination in the spinal cord white matter. Perforin-deficient mice demonstrated severely impaired MHC class I-restricted cytotoxicity against viral epitopes, but normal MHC class II-restricted delayed-type hypersensitivity responses to virus antigen. Despite demyelination, virus-infected perforin-deficient mice showed only minimal neurologic deficits as indicated by clinical disease score, activity monitoring, and footprint analysis. Perforin- and MHC class II-deficient mice (with functional CD8(+) T cells and perforin molecules and an H-2(b) haplotype) had comparable demyelination and genotype, however, only the latter showed severe clinical disease. Gld and lpr mice demonstrated normal TMEV-specific cytotoxicity and maintained resistance to TMEV-induced demyelinating disease. These studies implicate perforin release by CD8(+) T cells as a potential mechanism by which neurologic deficits are induced after demyelination. PMID- 9736652 TI - Embryonic expression of the myelin basic protein gene: identification of a promoter region that targets transgene expression to pioneer neurons. AB - The myelin basic protein (MBP) gene produces two families of structurally related proteins from three different promoters-the golli products, generated from the most upstream promoter, and the MBPs, produced from the two downstream promoters. In this report we describe the expression of golli proteins within some of the earliest neuronal populations of the brain, including Cajal-Retzius cells and preplate neurons of the forebrain, representing a new marker for these cells. To identify elements responsible for neuronal expression of the golli products, we generated transgenic animals from constructs containing different portions of the upstream promoter. A construct containing 1.1 kb immediately upstream of the golli transcription start site targeted expression of beta-galactosidase to preplate neurons and a subset of Cajal-Retzius cells in transgenic mice-the first reported genetic element to target expression to these pioneer cortical populations. Although expression in Cajal-Retzius cells declined with embryonic development, preplate cells continued to express the transgene after arriving at their final destination in the subplate. Interestingly, expression persisted in subplate neurons found within a distinct layer between the white matter and cortical layer VI well into postnatal life. Birth dating studies with bromodeoxyuridine indicated that these neurons were born between E10.5 and E12.5. Thus, the transgene marked subplate neurons from their birth, providing a fate marker for these cells. This work suggests a role for the MBP gene in the early developing brain long before myelination and especially in the pioneer cortical neurons important in the formation of the cortical layers. PMID- 9736653 TI - Synaptic competition during the reformation of a neuromuscular map. AB - We have been studying the mechanisms whereby pools of motor neurons establish a rostrocaudal bias in the position of their synapses in some skeletal muscles. The serratus anterior (SA) muscle of the rat displays a rostrocaudal topographic map before birth, and the topography is re-established after denervation. In this report, we explore the potential role of synaptic competition between innervating axons as a means of generating topographic specificity. We followed the progress of the reformation of this map in neonatal animals under conditions that enhanced the likelihood of observing synaptic competition. This was accomplished by forcing caudal axons to regenerate ahead of rostral axons onto a surgically reduced SA muscle. In this way, caudal (C7) motor neurons had unopposed access to vacated synaptic sites on the remaining rostral half of the SA before the return of the rostral (C6) axons. Intracellular recording revealed that 2 d after the second denervation, most of the reinnervated end plates contained only axons from the C7 branch; the remaining reinnervated end plates received input from C6 only or were multiply innervated by C6 and C7 axons. After 6 d, the pattern was reversed, with most end plates innervated exclusively by C6. After 17 d, axons from C6 were the sole input to reinnervated end plates. During the transition from C7- to C6-dominated input, at end plates coinnervated by C6 and C7 axons, the average quantal content from C6 was the same as that from C7; after 7 d, the quantal content of C6 was greater than that of C7. We have thus developed an experimental situation in which the outcome of synaptic competition is predictable and can be influenced by the positional labels associated with axons from different levels in the spinal cord. PMID- 9736654 TI - TrkB and TrkC signaling are required for maturation and synaptogenesis of hippocampal connections. AB - Recent studies have suggested a role for neurotrophins in the growth and refinement of neural connections, in dendritic growth, and in activity-dependent adult plasticity. To unravel the role of endogenous neurotrophins in the development of neural connections in the CNS, we studied the ontogeny of hippocampal afferents in trkB (-/-) and trkC (-/-) mice. Injections of lipophilic tracers in the entorhinal cortex and hippocampus of newborn mutant mice showed that the ingrowth of entorhinal and commissural/associational afferents to the hippocampus was not affected by these mutations. Similarly, injections of biocytin in postnatal mutant mice (P10-P16) did not reveal major differences in the topographic patterns of hippocampal connections. In contrast, quantification of biocytin-filled axons showed that commissural and entorhinal afferents have a reduced number of axon collaterals (21-49%) and decreased densities of axonal varicosities (8-17%) in both trkB (-/-) and trkC (-/-) mice. In addition, electron microscopic analyses showed that trkB (-/-) and trkC (-/-) mice have lower densities of synaptic contacts and important structural alterations of presynaptic boutons, such as decreased density of synaptic vesicles. Finally, immunocytochemical studies revealed a reduced expression of the synaptic associated proteins responsible for synaptic vesicle exocytosis and neurotransmitter release (v-SNAREs and t-SNAREs), especially in trkB (-/-) mice. We conclude that neither trkB nor trkC genes are essential for the ingrowth or layer-specific targeting of hippocampal connections, although the lack of these receptors results in reduced axonal arborization and synaptic density, which indicates a role for TrkB and TrkC receptors in the developmental regulation of synaptic inputs in the CNS in vivo. The data also suggest that the genes encoding for synaptic proteins may be targets of TrkB and TrkC signaling pathways. PMID- 9736655 TI - The survival-promoting effect of glial cell line-derived neurotrophic factor on axotomized corticospinal neurons in vivo is mediated by an endogenous brain derived neurotrophic factor mechanism. AB - Autocrine trophic functions of brain-derived neurotrophic factor (BDNF) have been proposed for many central neurons because this neurotrophin displays striking colocalization with its receptor trkB within the CNS. In the cortex, the distribution patterns of BDNF and trkB expression are almost identical. Corticospinal neurons (CSNs) are a major cortical long-distance projecting system. They are localized in layer V of the somatosensory cortex, and their axons project into the spinal cord where they contribute to the innervation of spinal motoneurons. We have shown recently that adult CSNs express trkB mRNA and are rescued from axotomy-induced death by BDNF treatment. Half of the axotomized CSNs survived without BDNF infusions. These findings raise the possibility that endogenous cortical BDNF is involved in the trophic support of this neuronal population. To test the hypothesis that endogenous cortical BDNF promotes survival of adult CSNs, we infused the BDNF-neutralizing affinity-purified antibody RAB to axotomized and unlesioned CSNs for 7 d. This treatment resulted in increased death of axotomized CSNs. Survival of unlesioned CSNs was not affected by RAB treatment. In situ hybridizations for BDNF and trkB mRNA revealed that virtually all CSNs express trkB, whereas only half of them express BDNF. Thus, autocrine/paracrine mechanisms are likely to contribute to the endogenous BDNF protection of axotomized CSNs. We have demonstrated previously that, in addition to BDNF, glial cell line-derived neurotrophic factor (GDNF) and neurotrophin 3 (NT-3) also rescue CSNs from axotomy-induced death. We now show that the rescuing by GDNF requires the presence of endogenous cortical BDNF, implicating a central role of this neurotrophin in the trophic support of axotomized CSNs and a trophic cross-talk between BDNF and GDNF regarding the maintenance of lesioned CSNs. In contrast, NT-3 promotes survival of axotomized CSNs even when endogenous cortical BDNF is neutralized by RAB, indicating a potential of compensatory mechanisms for the trophic support of CSNs. PMID- 9736656 TI - Cyclic AMP elevation is sufficient to promote the survival of spinal motor neurons in vitro. AB - The short-term survival of highly purified embryonic spinal motor neurons (SMNs) in culture can be promoted by many peptide trophic factors, including brain derived neurotrophic factor (BDNF), ciliary neurotrophic factor (CNTF), fibroblast growth factor (FGF), glial-derived neurotrophic factor (GDNF), and hepatocyte growth factor (HGF). We have asked whether these peptides are sufficient to promote the long-term survival of purified E15 SMNs. Contrary to previous reports, we find that when SMNs are cultured in serum-free medium containing a single peptide trophic factor only approximately one-third of the cells survive for 3 d in culture. When multiple factors are combined, additive effects on survival are observed transiently, but by 7 d of culture the majority of SMNs has died. Surprisingly, when cAMP levels are elevated, the majority of SMNs extend processes and survive for 1 week in culture in the absence of peptide trophic factors, even in low-density cultures. A combination of five peptide trophic factors, together with cAMP elevation, promotes the long-term survival of most of the SMNs in serum-free culture for 3 weeks. These findings provide useful culture conditions for studying the properties of SMNs and have implications for the treatment of motor neuron diseases. PMID- 9736657 TI - Reciprocal regulation of ciliary neurotrophic factor receptors and acetylcholine receptors during synaptogenesis in embryonic chick atria. AB - Ciliary neurotrophic factor (CNTF) has been implicated in the development, survival, and maintenance of a broad range of neurons and glia in the peripheral nervous system and the CNS. Evidence also suggests that CNTF may affect development of cells outside the nervous system. We have found that functional CNTF and its receptor are expressed in developing embryonic chick heart and may be involved in parasympathetic synapse formation. CNTF and CNTF receptor mRNA levels were highest at embryonic day 11 (E11)-E13, the period of parasympathetic innervation in chick atria. Levels of atrial CNTF receptor mRNA were fourfold greater at E13 than at E6 and at E13 were 2.5-fold higher in atria than in ventricle, corresponding to the higher degree of parasympathetic innervation occurring in atria. Treatment of isolated atria or cultured atrial myocytes with recombinant human or avian CNTF resulted in the tyrosine phosphorylation and nuclear translocation of the signal transducer and activator of transcription STAT3. The developmental increase in atrial CNTF receptor mRNA was enhanced by stimulating muscarinic receptors with carbachol in ovo and was inhibited by blocking muscarinic cholinergic receptors with atropine. Treatment of cultured atrial myocytes with CNTF resulted in a twofold increase in the levels of muscarinic receptors. Thus, CNTF was able to regulate a key component of parasympathetic synapses on atrial myocytes. These results suggest a postsynaptic role for CNTF in the onset of parasympathetic function in the developing heart and provide new clues to molecular mechanisms directing synapse formation at targets of the autonomic nervous system. PMID- 9736658 TI - Sensory processing in the pallium of a mormyrid fish. AB - To investigate the functional organization of higher brain levels in fish we test the hypothesis that the dorsal gray mantle of the telencephalon of a mormyrid fish has discrete receptive areas for several sensory modalities. Multiunit and compound field potentials evoked by auditory, visual, electrosensory, and water displacement stimuli in this weakly electric fish are recorded with multiple semimicroelectrodes placed in many tracks and depths in or near telencephalic area dorsalis pars medialis (Dm). Most responsive loci are unimodal; some respond to two or more modalities. Each modality dominates a circumscribed area, chiefly separate. Auditory and electrical responses cluster in the dorsal 500 micrometer of rostral and caudolateral Dm, respectively. Two auditory subdivisions underline specialization of this sense. Mechanoreception occupies a caudal area overlapping electroreception but centered 500 micrometer deeper. Visual responses scatter widely through ventral areas. Auditory, electrosensory, and mechanosensory responses are dominated by a negative wave within the first 50 msec, followed by 15-55 Hz oscillations and a slow positive wave with multiunit spikes lasting from 200 to 500 msec. Stimuli can induce shifts in coherence of certain frequency bands between neighboring loci. Every electric organ discharge command is followed within 3 msec by a large, mainly negative but generally biphasic, widespread corollary discharge. At certain loci large, slow ("deltaF") waves usually precede transient shifts in electric organ discharge rate. Sensory-evoked potentials in this fish pallium may be more segregated than in elasmobranchs and anurans and have some surprising similarities to those in mammals. PMID- 9736659 TI - Increase in serotonin-1A autoreceptors in the midbrain of suicide victims with major depression-postmortem evidence for decreased serotonin activity. AB - It has been hypothesized that a deficit in serotonin may be a crucial determinant in the pathophysiology of major depression. Serotonin-1A receptors are located on serotonin cell bodies in the midbrain dorsal raphe (DR) nucleus, and the activation of these receptors inhibits the firing of serotonin neurons and diminishes the release of this neurotransmitter in the prefrontal cortex. Repeated treatment with some antidepressant medications desensitizes serotonin-1A receptors in the rat midbrain. The present study determined whether the binding of [3H]8-hydroxy-2-(di-n-propyl)aminotetralin (8-OH-DPAT), an agonist at serotonin-1A receptors, is altered in the midbrain of suicide victims with major depression. Radiolabeling of the serotonin-1A receptor in the DR varied significantly along the rostral-to-caudal extent of the human midbrain. The binding of [3H]8-OH-DPAT to serotonin-1A receptors was increased significantly in the midbrain DR of suicide victims with major depression as compared with psychiatrically normal control subjects. In suicide victims with major depression, the increase in the binding of [3H]8-OH-DPAT to serotonin-1A receptors was detected in the entire DR and specifically localized to the dorsal and ventrolateral subnuclei. Enhanced radioligand binding of an agonist to inhibitory serotonin-1A autoreceptors in the human DR provides pharmacological evidence to support the hypothesis of diminished activity of serotonin neurons in suicide victims with major depression. PMID- 9736660 TI - A protein kinase, PKN, accumulates in Alzheimer neurofibrillary tangles and associated endoplasmic reticulum-derived vesicles and phosphorylates tau protein. AB - A possible role for a protein kinase, PKN, a fatty acid-activated serine/threonine kinase with a catalytic domain homologous to the protein kinase C family and a direct target for Rho, was investigated in the pathology of Alzheimer's disease (AD) using a sensitive immunocytochemistry on postmortem human brain tissues and a kinase assay for human tau protein. The present study provides evidences by light, electron, and confocal laser microscopy that in control human brains, PKN is enriched in neurons, where the kinase is concentrated in a subset of endoplasmic reticulum (ER) and ER-derived vesicles localized to the apical compartment of juxtanuclear cytoplasm, as well as late endosomes, multivesicular bodies, Golgi bodies, secretary vesicles, and nuclei. In AD-affected neurons, PKN was redistributed to the cortical cytoplasm and neurites and was closely associated with neurofibrillary tangles (NFTs) and their major constituent, abnormally modified tau. PKN was also found in degenerative neurites within senile plaques. In addition, we report that human tau protein is directly phosphorylated by PKN both in vitro and in vivo. Thus, our results suggest a specific role for PKN in NFT formation and neurodegeneration in AD damaged neurons. PMID- 9736661 TI - A statistical paradigm for neural spike train decoding applied to position prediction from ensemble firing patterns of rat hippocampal place cells. AB - The problem of predicting the position of a freely foraging rat based on the ensemble firing patterns of place cells recorded from the CA1 region of its hippocampus is used to develop a two-stage statistical paradigm for neural spike train decoding. In the first, or encoding stage, place cell spiking activity is modeled as an inhomogeneous Poisson process whose instantaneous rate is a function of the animal's position in space and phase of its theta rhythm. The animal's path is modeled as a Gaussian random walk. In the second, or decoding stage, a Bayesian statistical paradigm is used to derive a nonlinear recursive causal filter algorithm for predicting the position of the animal from the place cell ensemble firing patterns. The algebra of the decoding algorithm defines an explicit map of the discrete spike trains into the position prediction. The confidence regions for the position predictions quantify spike train information in terms of the most probable locations of the animal given the ensemble firing pattern. Under our inhomogeneous Poisson model position was a three to five times stronger modulator of the place cell spiking activity than theta phase in an open circular environment. For animal 1 (2) the median decoding error based on 34 (33) place cells recorded during 10 min of foraging was 8.0 (7.7) cm. Our statistical paradigm provides a reliable approach for quantifying the spatial information in the ensemble place cell firing patterns and defines a generally applicable framework for studying information encoding in neural systems. PMID- 9736662 TI - Where and when to pay attention: the neural systems for directing attention to spatial locations and to time intervals as revealed by both PET and fMRI. AB - Although attention is distributed across time as well as space, the temporal allocation of attention has been less well researched than its spatial counterpart. A temporal analog of the covert spatial orientation task [Posner MI, Snyder CRR, Davidson BJ (1980) Attention and the detection of signals. J Exp Psychol Gen 109:160-174] was developed to compare the neural systems involved in directing attention to spatial locations versus time intervals. We asked whether there exists a general system for allocating attentional resources, independent of stimulus dimension, or whether functionally specialized brain regions are recruited for directing attention toward spatial versus temporal aspects of the environment. We measured brain activity in seven healthy volunteers by using positron emission tomography (PET) and in eight healthy volunteers by using functional magnetic resonance imaging (fMRI). The task manipulated cued attention to spatial locations (S) and temporal intervals (T) in a factorial design. Symbolic central cues oriented subjects toward S only (left or right), toward T only (300 msec or 1500 msec), toward both S and T simultaneously, or provided no information regarding S or T. Subjects also were scanned during a resting baseline condition. Behavioral data showed benefits and costs for performance during temporal attention similar to those established for spatial attention. Brain-imaging data revealed a partial overlap between neural systems involved in the performance of spatial versus temporal orientation of attention tasks. Additionally, hemispheric asymmetries revealed preferential right and left parietal activation for spatial and temporal attention, respectively. Parietal cortex was activated bilaterally by attending to both dimensions simultaneously. This is the first direct comparison of the neural correlates of attending to spatial versus temporal cues. PMID- 9736663 TI - Critical role of axonal A-type K+ channels and axonal geometry in the gating of action potential propagation along CA3 pyramidal cell axons: a simulation study. AB - A model of CA3 pyramidal cell axons was used to study a new mode of gating of action potential (AP) propagation along the axon that depends on the activation of A-type K+ current (Debanne et al., 1997). The axonal membrane contained voltage-dependent Na+ channels, K+ channels, and A-type K+ channels. The density of axonal A-channels was first determined so that (1) at the resting membrane potential an AP elicited by a somatic depolarization was propagated into all axon collaterals and (2) propagation failures occurred when a brief somatic hyperpolarization preceded the AP induction. Both conditions were fulfilled only when A-channels were distributed in clusters but not when they were homogeneously distributed along the axon. Failure occurs in the proximal part of the axon. Conduction failure could be determined by a single cluster of A-channels, local decrease of axon diameter, or axonal elongation. We estimated the amplitude and temporal parameters of the hyperpolarization required for induction of a conduction block. Transient and small somatic hyperpolarizations, such as simulated GABAA inhibitory postsynaptic potentials, were able to block the AP propagation. It was shown that AP induction had to occur with a short delay (<30 msec) after the hyperpolarization. We discuss the possible conditions in which such local variations of the axon geometry and A-channel density may occur and the incidence of AP propagation failures on hippocampal network properties. PMID- 9736664 TI - Relationship between fos production and classical fear conditioning: effects of novelty, latent inhibition, and unconditioned stimulus preexposure. AB - The relationship between FOS production in the sensory cortex and limbic system and the ability of C57BL/6N mice to acquire context- and tone-dependent freezing were investigated after fear conditioning, which was achieved by exposure of mice to context only or context and tone (10 kHz, 75 dB) as conditioned stimuli (Cs) paired with an electric footshock (0.7 mA, constant) as unconditioned stimulus (Us). The effect of preexposure to Cs or Cs paired with Us on FOS production and learning was also tested. It was demonstrated that high simultaneous FOS production in the parietal cortex, hippocampus, and amygdala paralleled the ability of mice to acquire strong freezing responses to novel Cs. After contextual preexposure (latent inhibition), FOS production could be elicited in the central amygdala only by shock and in the basolateral amygdala only by tone. Under these conditions, the ability of mice to acquire contextual freezing was almost abolished, whereas tone-dependent freezing was reduced. Lacking FOS production in the central amygdala after preexposure to context followed by shock (Us preexposure effect) paralleled the inability of mice to acquire tone dependent freezing, although the tone elicited FOS production in the basolateral amygdala. On the basis of these findings it was concluded that synchronous Cs- and Us-induced FOS production in several defined forebrain areas was accompanied with associative learning of novel stimuli, and that a subsequent low level of FOS production might have been responsible or indicative for delayed conditioning to those stimuli. PMID- 9736665 TI - Regulation of hippocampal glucocorticoid receptor gene transcription and protein expression in vivo. AB - Glucocorticoid receptors (GRs) are glucocorticoid-activated transcription factors that modulate expression of a variety of neuronal genes. Appropriate control of GR expression is therefore critical for maintenance of cellular and organismic homeostasis. The present study assessed glucocorticoid regulation of the GR at the gene, mRNA, and protein level. Removal of circulating glucocorticoids (adrenalectomy) increased GR mRNA expression in CA1 and dentate gyrus (DG). Corticosterone (CORT) replacement normalized GR mRNA expression, whereas high doses slightly decreased GR mRNA in CA1. Parallel increases were observed using a probe complementary to the distal 3' untranslated region, indicating that mRNA changes were not attributable to selection of alternative polyadenylation site. Expression of a GR intronic sequence was also increased by adrenalectomy, consistent with increased gene transcription. Analysis of regional GR protein expression by immunoautoradiography did not reveal changes in GR protein in pyramidal cell layers; however, increased GR signal was seen in the stratum radiatum, indicating redistribution of GR to the cytosol. Western blot analysis confirmed adrenalectomy-induced increases in hippocampal GR levels. Administration of the mineralocorticoid receptor (MR) antagonist spironolactone increased both GR mRNA and protein in CA1 and DG, consistent with MR-mediated inhibition of GR transcription. However, high-dose CORT treatment did not decrease GR mRNA or protein levels. Chronic stress exposure did not downregulate GR mRNA or protein in hippocampus. The results suggest that the hippocampal GR is subject to heterologous regulation by the MR. In contrast, GR autoregulation is only evident during prolonged exposure to high-circulating glucocorticoid levels. PMID- 9736666 TI - Low doses of ethanol reduce evidence for nonlinear structure in brain activity. AB - Recent theories of the effects of ethanol on the brain have focused on its direct actions on neuronal membrane proteins. However, neuromolecular mechanisms whereby ethanol produces its CNS effects in low doses typically used by social drinkers (e.g., 2-3 drinks, 10-25 mM, 0.05-0.125 gm/dl) remain less well understood. We propose the hypothesis that ethanol may act by introducing a level of randomness or "noise" in brain electrical activity. We investigated the hypothesis by applying a battery of tests originally developed for nonlinear time series analysis and chaos theory to EEG data collected from 32 men who had participated in an ethanol/placebo challenge protocol. Because nonlinearity is a prerequisite for chaos and because we can detect nonlinearity more reliably than chaos, we concentrated on a series of measures that quantitated different aspects of nonlinearity. For each of these measures the method of surrogate data was used to assess the significance of evidence for nonlinear structure. Significant nonlinear structure was found in the EEG as evidenced by the measures of time asymmetry, determinism, and redundancy. In addition, the evidence for nonlinear structure in the placebo condition was found to be significantly greater than that for ethanol. Nonlinear measures, but not spectral measures, were found to correlate with a subject's overall feeling of intoxication. These findings are consistent with the notion that ethanol may act by introducing a level of randomness in neuronal processing as assessed by EEG nonlinear structure. PMID- 9736667 TI - Postnatal development of type I and type II hair cells in the mouse utricle: acquisition of voltage-gated conductances and differentiated morphology. AB - The type I and type II hair cells of mature amniote vestibular organs have been classified according to their afferent nerve terminals: calyx and bouton, respectively. Mature type I and type II cells also have different complements of voltage-gated channels. Type I cells alone express a delayed rectifier, gK,L, that is activated at resting potential. We report that in mouse utricles this electrophysiological differentiation occurs during the first postnatal week. Whole-cell currents were recorded from hair cells in denervated organotypic cultures and in acutely excised epithelia. From postnatal day 1 (P1) to P3, most hair cells expressed a delayed rectifier that activated positive to resting potential and a fast inward rectifier, gK1. Between P4 and P8, many cells acquired the type I-specific conductance gK,L and/or a slow inward rectifier, gh. By P8, the percentages of cells expressing gK,L and gh were at mature levels. To investigate whether the electrophysiological differentiation correlated with morphological changes, we fixed utricles at different times between P0 and P28. Ultrastructural criteria were developed to classify cells when calyces were not present, as in cultures and neonatal organs. The morphological and electrophysiological differentiation followed different time courses, converging by P28. At P0, when no hair cells expressed gK,L, 33% were classified as type I by ultrastructural criteria. By P28, approximately 60% of hair cells in acute preparations received calyx terminals and expressed gK,L. Data from the denervated cultures showed that neither electrophysiological nor morphological differentiation depended on ongoing innervation. PMID- 9736668 TI - Food restriction enhances the central rewarding effect of abused drugs. AB - Chronic food restriction increases the systemic self-administration and locomotor stimulating effect of abused drugs. However, it is not clear whether these behavioral changes reflect enhanced rewarding potency or a CNS-based modulatory process. The purpose of this study was to determine whether food restriction specifically increases the rewarding potency of drugs, as indexed by their threshold-lowering effect on lateral hypothalamic self-stimulation, and whether any such effect can be attributed to an enhanced central response rather than changes in drug disposition. When drugs were administered systemically, food restriction potentiated the threshold-lowering effect of amphetamine (0.125, 0.25, and 0.5 mg/kg, i.p.), phencyclidine (1.0, 2.0, and 3.0 mg/kg, i.p.), and dizocilpine (MK-801) (0.0125, 0.05, and 0.1 mg/kg, i.p.) but not nicotine (0.15, 0.3, 0.45 mg/kg, s.c.). When amphetamine (25.0, 50.0, and 100.0 microgram) and MK 801 (5.0, 10.0, and 20.0 microgram) were administered via the intracerebroventricular route, food restriction again potentiated the threshold lowering effects and increased the locomotor-stimulating effects of both drugs. These results indicate that food restriction increases the sensitivity of neural substrates for rewarding and stimulant effects of drugs. In light of work that attributes rewarding effects of MK-801 to blockade of NMDA receptors on medium spiny neurons in nucleus accumbens, the elements affected by food restriction may lie downstream from the mesoaccumbens dopamine neurons whose terminals are the site of amphetamine-rewarding action. Possible metabolic-endocrine triggers of this effect are discussed, as is the likelihood that mechanisms mediating the modulatory effect of food restriction differ from those mediating sensitization by intermittent drug exposure. PMID- 9736669 TI - Predicting the consequences of our own actions: the role of sensorimotor context estimation. AB - During self-generated movement it is postulated that an efference copy of the descending motor command, in conjunction with an internal model of both the motor system and environment, enables us to predict the consequences of our own actions (von Helmholtz, 1867; Sperry, 1950; von Holst, 1954; Wolpert, 1997). Such a prediction is evident in the precise anticipatory modulation of grip force seen when one hand pushes on an object gripped in the other hand (Johansson and Westling, 1984; Flanagan and Wing, 1933). Here we show that self-generation is not in itself sufficient for such a prediction. We used two robots to simulate virtual objects held in one hand and acted on by the other. Precise predictive grip force modulation of the restraining hand was highly dependent on the sensory feedback to the hand producing the load. The results show that predictive modulation requires not only that the movement is self-generated, but also that the efference copy and sensory feedback are consistent with a specific context; in this case, the manipulation of a single object. We propose a novel computational mechanism whereby the CNS uses multiple internal models, each corresponding to a different sensorimotor context, to estimate the probability that the motor system is acting within each context. PMID- 9736670 TI - Modulation of neuronal activity in superior colliculus by changes in target probability. AB - Complex visual scenes require that a target for an impending saccadic eye movement be selected from a larger number of possible targets. We investigated whether changing the probability that a visual stimulus would be selected as the target for a saccade altered activity of monkey superior colliculus (SC) neurons in two experiments. First, we changed the number of possible targets on each trial. Second, we kept the visual display constant and presented a single saccade target repeatedly so that target probability was established over time. Buildup neurons in the SC, those with delay period activity, showed a consistent reduction in activity as the probability of the saccade decreased, independent of the visual stimulus configuration. Other SC neurons, fixation and burst, were largely unaffected by the changes in saccade target probability. Because we had monkeys making saccades to many locations within the visual field, we could examine activity associated with saccades outside of the movement field of neurons. We found the activity of buildup neurons to be similar across the SC, before the target was identified, and reduced when the number of possible targets increased. The results of our experiments are consistent with a role for this activity in establishing a motor set. We found, consistent with this interpretation, that the activity of these neurons was predictive of the latency of a saccadic eye movement and not other saccade parameters such as end point or peak velocity. PMID- 9736671 TI - Distributed encoding and retrieval of spatial memory in the hippocampus. AB - To determine whether memory is processed in a localized or distributed manner by the hippocampus, we inactivated small regions of the structure in pretrained rats before a retention test. Ibotenic acid-induced lesions removing 40% of the hippocampal tissue disrupted retrieval of spatial memory in a water maze but failed to affect new learning or retrieval of a task that was acquired postoperatively. Partial inactivation of the hippocampus by local intrahippocampal 5-aminomethyl-3-hydroxyisoxazole muscimol infusion also impaired retrieval but not new learning. This impairment was temporary; infusions had no effect on retrieval of predrug performance when the test was conducted 48 hr after the infusion. Systematic variation of the volume of dorsal and ventral hippocampal lesions showed that successful retrieval required the integrity of the entire dorsal 70% of the hippocampus. Our data suggest that although spatial tasks can be acquired with local ensembles of hippocampal neurons when other parts of the hippocampus are inactivated, spatial memory is normally both encoded and retrieved by a widely distributed hippocampal network. PMID- 9736673 TI - Center-surround antagonism based on disparity in primate area MT. AB - Most neurons in primate visual area MT have a large, modulatory region surrounding their classically defined receptive field, or center. The velocity tuning of this "surround" is generally antagonistic to the center, making it potentially useful for detecting image discontinuities on the basis of differential motion. Because classical MT receptive fields are also disparity selective, one might expect to find disparity-based surround antagonism as well; this would provide additional information about image discontinuities. However, the effects of disparity in the MT surround have not been studied previously. We measured single-neuron responses to variable-disparity moving patterns in the MT surround while holding a central moving pattern at a fixed disparity. Of the 130 neurons tested, 84% exhibited a modulatory surround, and in 52% of these, responses were significantly affected by disparity in the surround. In most cases, disparity effects in the surround were antagonistic to the center; that is, neurons were generally suppressed when center and surround stimuli had the same disparity, with decreasing suppression as the center and surround stimuli became separated in depth. Also, the effects of disparity and direction were mainly additive; i.e., disparity effects were generally independent of direction, and vice versa. These results suggest that the MT center-surround apparatus provides information about image discontinuities, not only on the basis of velocity differences but on the basis of depth differences as well. This supports the hypothesis that MT surrounds have a role in image segmentation. PMID- 9736672 TI - Activity-dependent pH shifts and periodic recurrence of spontaneous interictal spikes in a model of focal epileptogenesis. AB - The mechanisms that control the periodicity of spontaneous epileptiform cortical potentials were investigated in the in vitro isolated guinea pig brain preparation. A brief intracortical application of bicuculline in the piriform cortex induced spontaneous interictal spikes (sISs) that recurred with high periodicity (8.5 +/- 3.1 sec, mean +/- SD). Intracellular recordings from principal neurons showed that the early phase of the inter-sIS period is caused by a GABAb receptor-mediated inhibitory potential. The late component of the interspike period correlated to a slowly decaying depolarization abolished at membrane potentials positive to -32.1 +/- 5.3 mV and was not associated with membrane conductance changes. Specific pharmacological tests excluded the contribution of synaptic and intrinsic conductances to the late inter-sIS interval. Recordings with ion-sensitive electrodes demonstrated that sISs determined both a rapid increase in extracellular K+ concentration (0.5-1 mM) and an extracellular alkalinization (0.05-0.08 pH units) that slowly decayed during the inter-sIS period and returned to control values just before a subsequent sIS was generated. These observations were not congruous with the presence of a silent period, because both extracellular increase in K+ and alkalinization are commonly associated with an increase in neuronal excitability. Extracellular alkalinization could be correlated to an sIS-induced intracellular acidification, a phenomenon that reduces cell coupling by impairing gap junction function. When intracellular acidification was transiently prevented by arterial perfusion with NH4Cl (10-20 mM), spontaneous ictal-like epileptiform discharges were induced. In addition, the gap junction blockers octanol (0.2-2 mM) and 18-alpha glycyrrethinic acid (20 microM) applied either via the arterial system or locally in the cortex completely and reversibly abolished the sIS. The results reported here suggest that the massive cell discharge associated with an sIS induce a strong inhibition, possibly secondary to a pH-dependent uncoupling of gap junctions, that regulates sIS periodicity. PMID- 9736674 TI - Cognitive channels computing action distance and direction. AB - Visually guided, goal-directed reaching requires encoding action distance and direction from attributes of visual landmarks. We identified a cognitive mechanism that seemingly performs visual motor extension before action initiation and replicated and extended previous results that identified a mechanism for visual motor mental rotation. We find that humans systematically delay action onset while newly planning increasingly distant arm movements beyond a visual landmark, consistent with an internal representation for visual motor extension. Onset times also changed systematically during concurrent mental rotation and visual motor extension computations required to process new directions and distances. Visual motor extension associated with reaching slowed when participants needed to plan action direction within the same time frame, whereas mental rotation efficiency was unaffected by concurrent needs to prepare action distance. In contrast to parallel direction and distance computations needed for direct aiming to a visual target, the planning of new directions and distances likely occurs at distinct times. When considered with previous findings, the current results suggest the existence of an intermediate component of motor preparation that engages a covert mechanism of cognitive motor planning. PMID- 9736675 TI - Inflammatory mediators sensitize acutely axotomized nerve fibers to mechanical stimulation in the rat. AB - Many axotomized myelinated as well as unmyelinated cutaneous nerve fibers are sensitive to mechanical stimuli applied to the cut nerve end within a few hours after nerve lesion. Here we investigated the influence of inflammatory mediators on this ectopic mechanosensitivity after cutting and ligating the sural nerve in anesthetized rats. Neural activity was recorded from single axons in filaments teased from the sural or sciatic nerve proximally to the lesion site 2-33 hr after axotomy. Using calibrated von Frey hairs (1.0-128.5 mN), 30 sec trains of phasic stimuli were applied to the cut nerve end immediately before and after local application of a mixture of inflammatory mediators [inflammatory soup (IS), consisting of bradykinin, 5-HT, prostaglandin E2, histamine (all 10 microM), and K+ 7 mM, pH 7.0] for 2 min. Before as well as after IS application, von Frey thresholds were significantly lower in myelinated (A) fibers than in unmyelinated (C) fibers. IS application enhanced the ectopic mechanical excitability, as expressed in reduced von Frey thresholds and increased response magnitudes, of most severed mechanosensitive C fibers (77%) and some mechanosensitive A fibers (46%). The sensitization lasted for 10-40 min after a 2 min IS application. Additionally, among axotomized nerve fibers unresponsive to probing of the nerve lesion site before IS application, 1 of 63 (1.6%) A and 3 of 106 (2.8%) C fibers became mechanosensitive immediately after IS application. The results indicate that after axotomy, inflammatory processes augment touch-evoked ectopic activity in lesioned sensory nerve fibers. Because many affected afferents are presumably of nociceptive function, their enhanced neural barrage may contribute to neuropathic pain states. PMID- 9736676 TI - Phasic firing time locked to cocaine self-infusion and locomotion: dissociable firing patterns of single nucleus accumbens neurons in the rat. AB - The activity of single nucleus accumbens (NAcc) neurons of rats was extracellularly recorded during intravenous cocaine self-administration sessions (0.7 mg/kg per infusion, fixed ratio 1). We reported previously that NAcc neurons showed a change, usually a decrease, in firing rate during the first 1 min after the cocaine-reinforced lever press. This postpress change was followed by a progressive reversal of that change, which began within the first 2 min after the press and was not complete until the last 1 min before the next lever press (termed the change + progressive reversal firing pattern). In the present study we documented a regular pattern of locomotion that occurred in parallel with the change + progressive reversal firing pattern. This observation suggested that discharges time locked to locomotion may determine the change + progressive reversal firing pattern. However, 55% of the neurons failed to show firing time locked to locomotion that could have contributed to the change + progressive reversal firing pattern. Moreover, for all neurons, the change + progressive reversal firing pattern was apparent even if the calculation of firing rate excluded all periods of locomotion. The present data showed that the change + progressive reversal firing pattern is not solely attributable to phasic changes in firing time locked to the execution of locomotion. The change + progressive reversal firing pattern closely mirrors changes in drug level and dopamine overflow observed by previous researchers and may thus be a component of the neurophysiological mechanism by which drug level regulates drug-taking behavior during an ongoing self-administration session. PMID- 9736678 TI - Introduction to Frontiers of Science. PMID- 9736677 TI - Parallel projection of amplitude and phase information from the hindbrain to the midbrain of the African electric fish Gymnarchus niloticus. AB - Two distinct sensory cues in electrosensory signals, amplitude modulation and differential phase modulation, are essential for an African wave-type electric fish, Gymnarchus, to perform the jamming avoidance responses. Individual neurons in the first brain station for central processing, the electrosensory lateral line lobe (ELL), were investigated by the in vivo whole-cell recording and labeling technique for their physiological responses, location, morphology, and projection areas. Neurons in the dorsal zone of the ELL responded selectively to amplitude modulation. Neurons in the outer cell layer of the medial zone were categorized physiologically into two groups: amplitude-sensitive and differential phase-sensitive. All but one neuron in the inner cell layer of the medial zone responded exclusively to differential phase modulation. All neurons recorded and labeled in the ELL had pyramidal morphology with large and extensive apical dendrites and less extensive basal dendrites. They were found to project to two midbrain nuclei: the nucleus praeeminentialis and the torus semicircularis. Amplitude-sensitive neurons in the dorsal zone projected exclusively to the lateral posterior subdivision, the torus semicircularis. Neurons in the medial zone projected to the medial dorsal and lateral anterior subdivisions of the torus semicircularis. Although some neurons in the ELL responded to both amplitude and differential phase modulation, they did not differentiate between temporal patterns of the two cues that encode necessary information for the jamming avoidance response. Overlapping projection of amplitude and differential phase-sensitive neurons to the torus semicircularis suggests integration of the two sensory cues in this nucleus. PMID- 9736679 TI - Meteorite impact and the mass extinction of species at the Cretaceous/Tertiary boundary. PMID- 9736680 TI - Prions. PMID- 9736681 TI - Quantum computing. PMID- 9736682 TI - Aging, life span, and senescence. PMID- 9736684 TI - String theory. PMID- 9736683 TI - Chemical physics of protein folding. PMID- 9736685 TI - New developments in the biology and treatment of HIV. PMID- 9736686 TI - Default taxonomy: Ernst Mayr's view of the microbial world. AB - This perspective is a response to a taxonomic proposal by E. Mayr ["Two empires or three?" (1998) Proc. Natl. Acad. Sci. USA 95, 9720-9723]. Mayr has suggested that the now accepted classification of life into three primary domains, Archaea, Bacteria, and Eucarya-originally proposed by myself and others--be abandoned in favor of the earlier Prokaryote-Eukaryote classification. Although the matter appears a taxonomic quibble, it is not that simple. At issue here are differing views as to the nature of biological classification, which are underlain by differing views as to what biology is and will be--matters of concern to all biologists. PMID- 9736687 TI - Synthesis, biophysical properties, and nuclease resistance properties of mixed backbone oligodeoxynucleotides containing cationic internucleoside guanidinium linkages: deoxynucleic guanidine/DNA chimeras. AB - The synthesis of mixed backbone oligodeoxynucleotides (18-mers) consisting of positively charged guanidinium linkages along with negatively charged phosphodiester linkages is carried out. The use of a base labile-protecting group for guanidinium linkage offers a synthetic strategy similar to standard oligonucleotide synthesis. The nuclease resistance of the oligodeoxyribonucleotides capped with guanidinium linkages at 5' and 3' ends are reported. The hybridization properties and sequence specificity of binding of these deoxynucleic guanidine/DNA chimeras with complementary DNA or RNA are described. PMID- 9736688 TI - Complex behavior of self-propagating reaction waves in heterogeneous media. AB - Self-propagating high temperature reaction waves, leading to the synthesis of advanced materials, are investigated in a variety of heterogeneous reaction systems by using a digital high-speed microscopic video recording technique. It is shown that, although on the macroscopic length and time scales, the reaction appears to move in a steady mode, on the microscopic level it has a complex character that is related to the reaction mechanism. PMID- 9736689 TI - Molecular determinants of bacterial adhesion monitored by atomic force microscopy. AB - Bacterial adhesion and the subsequent formation of biofilm are major concerns in biotechnology and medicine. The initial step in bacterial adhesion is the interaction of cells with a surface, a process governed by long-range forces, primarily van der Waals and electrostatic interactions. The precise manner in which the force of interaction is affected by cell surface components and by the physiochemical properties of materials is not well understood. Here, we show that atomic force microscopy can be used to analyze the initial events in bacterial adhesion with unprecedented resolution. Interactions between the cantilever tip and confluent monolayers of isogenic strains of Escherichia coli mutants exhibiting subtle differences in cell surface composition were measured. It was shown that the adhesion force is affected by the length of core lipopolysaccharide molecules on the E. coli cell surface and by the production of the capsular polysaccharide, colanic acid. Furthermore, by modifying the atomic force microscope tip we developed a method for determining whether bacteria are attracted or repelled by virtually any biomaterial of interest. This information will be critical for the design of materials that are resistant to bacterial adhesion. PMID- 9736690 TI - The 2.8-A structure of rat liver F1-ATPase: configuration of a critical intermediate in ATP synthesis/hydrolysis. AB - During mitochondrial ATP synthesis, F1-ATPase-the portion of the ATP synthase that contains the catalytic and regulatory nucleotide binding sites-undergoes a series of concerted conformational changes that couple proton translocation to the synthesis of the high levels of ATP required for cellular function. In the structure of the rat liver F1-ATPase, determined to 2.8-A resolution in the presence of physiological concentrations of nucleotides, all three beta subunits contain bound nucleotide and adopt similar conformations. This structure provides the missing configuration of F1 necessary to define all intermediates in the reaction pathway. Incorporation of this structure suggests a mechanism of ATP synthesis/hydrolysis in which configurations of the enzyme with three bound nucleotides play an essential role. PMID- 9736692 TI - Molecular dynamics study displays near in-line attack conformations in the hammerhead ribozyme self-cleavage reaction. AB - We have performed molecular dynamics (MD) calculations by using one of the recently solved crystal structures of a hammerhead ribozyme. By rotating the alpha, beta, gamma, delta, epsilon, and zeta torsion angles of the phosphate linkage of residue 17, the nucleobase at the cleavage site was slightly rotated out of the active site toward the solution. Unconstrained MD simulations exceeding 1 ns were performed on this starting structure solvated in water with explicit counter ions and two Mg2+ ions at the active site. Our results reveal that near attack conformations consistently were formed in the simulation. These near attack conformations are characterized by assumption of the 2'-hydroxyl to a near in-line position for attack on the -O-(PO2-)-O- phosphorous. Also during the time course of the MD study, one Mg2+ moved immediately to associate with a pro-R phosphate oxygen in the conserved core region, and the second Mg2+ remained associated with the pro-R oxygen on the phosphate linkage undergoing hydrolysis. These results are in accord with a one-metal ion mechanism of catalysis and give insight into the possible roles of many of the conserved residues in the ribozyme. PMID- 9736691 TI - Base pair switching by interconversion of sugar puckers in DNA extended by proteins of RecA-family: a model for homology search in homologous genetic recombination. AB - Escherichia coli RecA is a representative of proteins from the RecA family, which promote homologous pairing and strand exchange between double-stranded DNA and single-stranded DNA. These reactions are essential for homologous genetic recombination in various organisms. From NMR studies, we previously reported a novel deoxyribose-base stacking interaction between adjacent residues on the extended single-stranded DNA bound to RecA protein. In this study, we found that the same DNA structure was induced by the binding to Saccharomyces cerevisiae Rad51 protein, indicating that the unique DNA structure induced by the binding to RecA-homologs was conserved from prokaryotes to eukaryotes. On the basis of this structure, we have formulated the structure of duplex DNA within filaments formed by RecA protein and its homologs. Two types of molecular structures are presented. One is the duplex structure that has the N-type sugar pucker. Its helical pitch is approximately 95 A (18.6 bp/turn), corresponding to that of an active, or ATP-form of the RecA filament. The other is one that has the S-type sugar pucker. Its helical pitch is approximately 64 A (12.5 bp/turn), corresponding to that of an inactive, or ADP-form of the RecA filament. During this modeling, we found that the interconversion of sugar puckers between the N type and the S-type rotates bases horizontally, while maintaining the deoxyribose base stacking interaction. We propose that this base rotation enables base pair switching between double-stranded DNA and single-stranded DNA to take place, facilitating homologous pairing and strand exchange. A possible mechanism for strand exchange involving DNA rotation also is discussed. PMID- 9736693 TI - Delta5-androstenediol is a natural hormone with androgenic activity in human prostate cancer cells. AB - It is known that androst-5-ene-3beta,17beta-diol (Adiol), a precursor of testosterone (T), can activate estrogen target genes. The androgenic activity of Adiol itself, however, is poorly understood. Using a transient transfection assay, we here demonstrate in human prostate cancer cells that Adiol can activate androgen receptor (AR) target genes in the presence of AR, and that AR coactivator ARA70 can further enhance this Adiol-induced AR transcriptional activity. In contrast to this finding, an active metabolite of dehydroepiandrosterone, 7-oxo-dehydroepiandrosterone, does not activate AR target gene in the absence or presence of ARA70. Thin layer chromatography analysis reveals that T, dihydrotestosterone, and 17beta-estradiol are undetectable in human prostate cancer DU145 cells after treatment with Adiol. Additionally, a proteolysis assay shows that a distinct ligand-receptor conformational difference exists between T-AR and Adiol-AR. Together, the above findings and the fact that T, but not Adiol, can induce transcriptional activity in a mutant AR (mtAR708), suggest that, without being metabolized into T, Adiol itself may represent a natural hormone with androgenic activity in human prostate cancer cells. Because two potent antiandrogens, hydroxyflutamide (Eulexin), and bicalutamide (casodex), that are widely used for the treatment of prostate cancer, fail to block Adiol mediated induction of AR transcriptional activity in prostate cancer cells, the effectiveness of so-called "total androgen blockage," a standard treatment for prostate cancer, may need to be reevaluated. PMID- 9736694 TI - Proteolysis of human eukaryotic translation initiation factor eIF4GII, but not eIF4GI, coincides with the shutoff of host protein synthesis after poliovirus infection. AB - Eukaryotic initiation factor (eIF) 4GI is a component of the cap-binding protein complex eIF4F, which is required for cap-dependent translation. Infection of cells by poliovirus results in a precipitous decline of host cell protein synthesis, which is preceded by the cleavage of eIF4GI. Cleavage of eIF4GI results in the inhibition of cap-dependent translation. Poliovirus translation is not affected by eIF4GI cleavage, however, because poliovirus mRNA is translated by a cap-independent mechanism. Cleavage of eIF4GI alone cannot explain the shutoff of host protein synthesis, because after infection in the presence of inhibitors of virus replication, eIF4GI is cleaved, yet host protein synthesis is only partially inhibited. Here we show that eIF4GII, a recently discovered functional homolog of eIF4GI, is more resistant to poliovirus-mediated cleavage than eIF4GI, and that its proteolysis is concomitant with the shutoff of host cell protein synthesis. Moreover, infection with poliovirus in the presence of inhibitors of virus replication resulted in efficient cleavage of eIF4GI, but only partial proteolysis of eIF4GII. Thus, cleavage of both eIF4GI and eIF4GII appears to be required for the shutoff of host protein synthesis after poliovirus infection. These results explain several earlier reports documenting the lack of correlation between eIF4GI cleavage and inhibition of cellular mRNA translation after poliovirus infection. PMID- 9736695 TI - Increase in the 64-kDa subunit of the polyadenylation/cleavage stimulatory factor during the G0 to S phase transition. AB - The amount of the 64-kDa subunit of polyadenylation/cleavage stimulatory factor (CstF-64) increases 5-fold during the G0 to S phase transition and concomitant proliferation induced by serum in 3T6 fibroblasts. Higher levels of CstF-64 result in an increase in CstF trimer. The rise in CstF-64 occurs at a time when the amount of poly(A)-containing RNA rose at least 5-8 fold in the cytoplasm. Primary human splenic B cells, resting in G0, show a similar 5-fold increase in CstF-64 when cultured under conditions inducing proliferation (CD40 ligand exposure). Therefore, the increase in CstF-64 is associated with the G0 to S phase transition. As B cell development progresses, RNA processing changes occur at the Ig heavy chain locus resulting in a switch from the membrane- to the upstream secretory-specific poly(A) site. Treating resting B cells with agents triggering this switch in Ig mRNA production along with proliferation (CD40 ligand plus lymphokines or Staphylococcus aureus protein A) induces no further increase in CstF-64 above that seen for proliferation alone. The rise in CstF-64 is therefore insufficient to induce secretion. After stimulation of a continuously growing B cell line with lymphokines, a switch to Ig micrometer secretory mRNA and protein occurs but without a change in the CstF-64 level. Therefore, an increase in CstF-64 levels is not necessary to mediate the differentiation-induced switch to secreted forms of Ig-micrometer heavy chain. Because augmentation of CstF-64 levels is neither necessary nor sufficient for Ig secretory mRNA production, we conclude that other lymphokine-induced factors play a role. PMID- 9736696 TI - Effects of transition metals on nitric oxide synthase catalysis. AB - The biosynthesis of nitric oxide (NO) by the enzyme NO synthase (NOS) proceeds by the hydroxylation of L-arginine to form NG-hydroxy-L-arginine followed by the conversion of NG-hydroxy-L-arginine to L-citrulline and NO. The previously identified requirements of this relatively complicated reaction include several protein-bound cofactors: cytochrome P450-type heme, flavin mononucleotide (FMN), flavin adenine dinucleotide (FAD), and tetrahydrobiopterin (H4B). In addition to L-arginine, NOS also requires the substrates NADPH and molecular oxygen. The role of H4B in NOS catalysis has long been a subject of debate and uncertainty fueled, in part, by the failure to detect any dependence of the NOS reaction on nonheme iron, a cofactor integral to catalysis in every other H4B-dependent enzyme. Here we report the ability of NOS to bind transition metals stoichiometrically, and demonstrate that the rate of catalysis is enhanced by nonheme iron. We also show that other divalent transition metals, including Cu, Zn, Co, and Ni, inhibit NOS catalysis. Also, the addition of Cu2+ to NOS inhibits heme reduction, whereas the addition of Fe2+ does not. Overall, the results appear to connect NOS to the known H4B/nonheme iron-dependent hydroxylases, and suggest a similar, if not identical, step in the NOS reaction mechanism. PMID- 9736697 TI - Rapid inhibition of interleukin-6 signaling and Stat3 activation mediated by mitogen-activated protein kinases. AB - Gene activation and cellular differentiation induced by interleukin-6 (IL-6) and transcription factor Stat3 are suppressed by several factors, including ionomycin, granulocyte/macrophage-colony-stimulating factor, and phorbol 12 myristate 13-acetate (PMA), that block IL-6-induced Stat3 activation. These inhibitory agents activate mitogen activated protein kinases (MAPKs), and thus the role of MAPKs in the mechanism of inhibition of Stat3 activation was investigated. Inhibition of IL-6-induced Stat3 activation by PMA and ionomycin was rapid (within 5 min) and did not require new RNA or protein synthesis. Inhibition of Stat3 DNA-binding activity and tyrosine phosphorylation by PMA, ionomycin, and granulocyte/macrophage-colony-stimulating factor was reversed when activation of the extracellular signal-regulated kinase (ERK) group of MAPKs was blocked by using specific kinase inhibitors. Expression of constitutively active MEK1, the kinase that activates ERKs, or overexpression of ERK2, but not JNK1, inhibited Stat3 activation. Inhibition of Stat3 correlated with suppression of IL 6-induction of a signal transducer and activator of transcription (STAT) dependent reporter gene. In contrast to IL-6, activation of Stat3 by interferon alpha was not inhibited. MEKs and ERKs inhibited IL-6 activation of Stat3 harboring a mutation at serine-727, the major site for serine phosphorylation, similar to inhibition of wild-type Stat3, and inhibited Janus kinases Jak1 and Jak2 upstream of Stat3 in the Jak-STAT-signaling pathway. These results demonstrate an ERK-mediated mechanism for inhibiting IL-6-induced Jak-STAT signaling that is rapid and inducible, and thus differs from previously described mechanisms for downmodulation of the Jak-STAT pathway. This inhibitory pathway provides a molecular mechanism for the antagonism of Stat3-mediated IL-6 activity by factors that activate ERKs. PMID- 9736698 TI - Indirect mutagenesis by oxidative DNA damage: formation of the pyrimidopurinone adduct of deoxyguanosine by base propenal. AB - Oxidation of endogenous macromolecules can generate electrophiles capable of forming mutagenic adducts in DNA. The lipid peroxidation product malondialdehyde, for example, reacts with DNA to form M1G, the mutagenic pyrimidopurinone adduct of deoxyguanosine. In addition to free radical attack of lipids, DNA is also continuously subjected to oxidative damage. Among the products of oxidative DNA damage are base propenals. We hypothesized that these structural analogs of malondialdehyde would react with DNA to form M1G. Consistent with this hypothesis, we detected a dose-dependent increase in M1G in DNA treated with calicheamicin and bleomycin, oxidizing agents known to produce base propenal. The hypothesis was proven when we determined that 9-(3-oxoprop-1-enyl)adenine gives rise to the M1G adduct with greater efficiency than malondialdehyde itself. The reactivity of base propenals to form M1G and their presence in the target DNA suggest that base propenals derived from oxidative DNA damage may contribute to the mutagenic burden of a cell. PMID- 9736699 TI - High-affinity binding of hemimethylated oriC by Escherichia coli membranes is mediated by a multiprotein system that includes SeqA and a newly identified factor, SeqB. AB - The binding of hemimethylated oriC to Escherichia coli membranes has been implicated in the prevention of premature reinitiation at newly replicated chromosomal origins in a reaction that involves the SeqA protein. We describe the resolution of the membrane-associated oriC-binding activity into two fractions, both of which are required for the high-affinity binding of hemimethylated oriC. The active component in one fraction is identified as SeqA. The active component of the second fraction is a previously undescribed protein factor, SeqB. The reconstituted system reproduced the salient characteristics of the membrane associated binding activity, suggesting that the membrane-associated oriC-binding machinery of E. coli is likely to be a multiprotein system that includes the SeqA and SeqB proteins. PMID- 9736700 TI - A novel yeast protein influencing the response of RNA polymerase II to transcriptional activators. AB - A sensitive in vitro crosslinking technique using a photoactive derivative of the chimeric activator LexA-E2F-1 was used to identify yeast proteins that might influence the response of RNA polymerase II to transcriptional activators. We found that a novel yeast protein, Xtc1p, could be covalently crosslinked to the activation domain of LexA-E2F-1 when this derivatized activator was bound to DNA upstream of an activator-responsive RNA polymerase II promoter. Because affinity chromatography experiments showed that Xtc1p also bound directly and specifically to the activation domains of E2F-1, the viral activator VP16, and the yeast activator Gal4p and copurified with the RNA polymerase II holoenzyme complex, Xtc1p may modulate the response of RNA polymerase II to multiple activators. Consistent with this notion, yeast strains deleted for the XTC1 gene exhibited pleiotropic growth defects, including temperature sensitivity, galactose auxotrophy, and a heightened sensitivity to activator overexpression, as well as an altered response to transcriptional activators in vivo. PMID- 9736701 TI - Simultaneous formation of functional leading and lagging strand holoenzyme complexes on a small, defined DNA substrate. AB - The biochemical characterization of leading and lagging strand DNA synthesis by bacteriophage T4 replication proteins has been addressed utilizing a small, defined primer/template. The ATP hydrolysis activity of 44/62, the clamp loading complex responsible for holoenzyme assembly, was monitored during assembly of both the leading and lagging strand holoenzyme complex. The ATPase activity of 44/62 diminishes once a functional holoenzyme is assembled on both the leading and lagging strand. The assembly of the lagging strand holoenzyme is facilitated by several factors including biotinylated streptavidin blocks at the end of the fork strands, preassembly of the leading strand holoenzyme, and by the presence of the DNA primase with ribonucleoside triphosphates. The resultant minimal replicative complex consists of two holoenzymes and a primase nested on a model replication fork derived from a 62-mer template/34-mer primer/36-mer lagging strand in an apparent 2:2:1:1 ratio of 45 protein:polymerase:primase:forked DNA. The 44/62 protein complex does not remain associated with the complex. The primase alone slowly synthesizes pentaribonucleotides on the forked DNA when the lagging strand contains a nonannealed TTG initiation site with the rate of synthesis greatly stimulated by the addition of the 41 helicase. The addition of deoxy-NTPs to this complex results in leading strand synthesis, but extension of the synthesized RNA primer does not occur. DNA synthesis in both the leading and lagging strand directions is achieved, however, when a 6-mer DNA primer is annealed to the primase recognition site of the forked DNA substrate. A model is presented that describes how leading and lagging strand DNA synthesis might be coordinated as well as the associated molecular interactions of the replicative proteins. PMID- 9736702 TI - Existence of distinct tyrosylprotein sulfotransferase genes: molecular characterization of tyrosylprotein sulfotransferase-2. AB - Tyrosylprotein sulfotransferase (TPST) is a 54- to 50-kDa integral membrane glycoprotein of the trans-Golgi network found in essentially all tissues investigated, catalyzing the tyrosine O-sulfation of soluble and membrane proteins passing through this compartment. Here we describe (i) an approach to identify the TPST protein, referred to as MSC (modification after substrate crosslinking) labeling, which is based on the crosslinking of a substrate peptide to TPST followed by intramolecular [35S]sulfate transfer from the cosubstrate 3' phosphoadenosine 5'-phosphosulfate (PAPS); and (ii) the molecular characterization of a human TPST, referred to as TPST-2, whose sequence is distinct from that reported [TPST-1; Ouyang, Y.-B., Lane, W. S. & Moore, K. L. (1998) Proc. Natl. Acad. Sci. USA 95, 2896-2901] while this study was in progress. Human TPST-2 is a type II transmembrane protein of 377 aa residues that is encoded by a ubiquitously expressed 1.9-kb mRNA originating from seven exons of a gene located on chromosome 22 (22q12.1). A 304-residue segment in the luminal domain of TPST-2 shows 75% amino acid identity to the corresponding segment of TPST-1, including conservation of the residues implicated in the binding of PAPS. Expression of the TPST-2 cDNA in CHO cells resulted in an approximately 13-fold increase in both TPST protein, as determined by MSC labeling, and TPST activity. A predicted 359-residue type II transmembrane protein in Caenorhabditis elegans with 45% amino acid identity to TPST-2 in a 257 residue segment of the luminal domain points to the evolutionary conservation of the TPST protein family. PMID- 9736703 TI - The natural polyamine spermine functions directly as a free radical scavenger. AB - The polyamines are small organic cations that are absolutely required for eukaryotic cell growth. Although their growth requirements are well established, the molecular functions of the polyamines are ill-defined. Oxidative damage to DNA by reactive oxygen species is a continual problem that cells must guard against to survive. The polyamine spermine, which is normally found in millimolar concentrations in the nucleus, is shown here to function directly as a free radical scavenger, and adducts formed as a result of this function are identified. These data suggest that spermine is a major natural intracellular compound capable of protecting DNA from free radical attack. PMID- 9736704 TI - Unexpected frameshifts from gene to expressed protein in a phage-displayed peptide library. AB - A library of long peptides displayed on the pIII protein of filamentous phage was used in biopanning experiments against several protein targets. We find that a large percentage of phage clones that bind specifically to a target contain peptide-encoding genes that do not have an ORF. Instead, the reading frame is either interrupted by one or more nonsuppressed stop codons, or a post transcriptional frameshift is needed to account for the expression of the minor phage coat protein pIII. The percentage of frameshifted clones varies depending on the target. It can be as high as 90% for clones specific for soluble forms of certain cytokine receptors. Conversely, biopanning against four mAbs did not yield any frameshifted clones. Our studies focused on one clone that binds specifically to rat growth hormone binding protein (GHBP) yet does not have an ORF. A secondary peptide library containing random mutations of this sequence was constructed and panned against GHBP to optimize and correct the reading frame. In the last round (round two) of panning with this library, none of the phage clones that bound to GHBP had an ORF. However, careful analysis of these clones allowed us to design a synthetic peptide capable of binding to GHBP. The results of this study indicate that ORFs are not required to obtain gene expression of the minor coat protein of filamentous phage and suggest that some ORF- clones may have a selective advantage over the clones having ORFs. PMID- 9736705 TI - Ciona intestinalis nuclear receptor 1: a member of steroid/thyroid hormone receptor family. AB - Nuclear hormone receptors comprise a large family of zinc finger transcription factors, some with hydrophobic ligands, such as thyroid hormone, vitamin D, steroids, etc., and others for which no ligand has been found. Thyroid hormone receptors (TRs) generally are considered to be confined to the vertebrata that possess a thyroid gland. Tunicates represent the most primitive of the chordates, and there are data supporting a role for thyroid hormone in their metamorphosis, but no data are available on TRs in this genus; hence, we have studied Ciona intestinalis. Screening of a Ciona library with the DNA binding domain of Xenopus laevis TR (xTR) resulted in the isolation of a nuclear hormone receptor, C. intestinalis nuclear receptor 1 (CiNR1). CiNR1 is similar to TRs of more evolved species with a conserved DNA binding domain whereas the ligand binding domain shows poor homology to vertebrate sequences. The C-terminal part of CiNR1 spans approximately 200 amino acids more than other TRs, lacks the AF2 transactivation domain, and is not able to bind triiodothyronine. Phylogenetically, CiNR1 appears to be close to the common ancestral gene of TRs. Expression of CiNR1 was limited to the developing embryo and the larval stage, which suggests a role during development and metamorphosis. In transfection experiments, CiNR1 down-regulated basal transcription of a reporter gene driven by the TR palindrome responsive element. When CiNR1 was cotransfected with chicken TRalpha, it attenuated the normal thyroid hormone response in a dominant negative fashion. This attenuation required the C-terminal portion of the molecule. PMID- 9736706 TI - Clustering of low-energy conformations near the native structures of small proteins. AB - Recent experimental studies of the denatured state and theoretical analyses of the folding landscape suggest that there are a large multiplicity of low-energy, partially folded conformations near the native state. In this report, we describe a strategy for predicting protein structure based on the working hypothesis that there are a greater number of low-energy conformations surrounding the correct fold than there are surrounding low-energy incorrect folds. To test this idea, 12 ensembles of 500 to 1,000 low-energy structures for 10 small proteins were analyzed by calculating the rms deviation of the Calpha coordinates between each conformation and every other conformation in the ensemble. In all 12 cases, the conformation with the greatest number of conformations within 4-A rms deviation was closer to the native structure than were the majority of conformations in the ensemble, and in most cases it was among the closest 1 to 5%. These results suggest that, to fold efficiently and retain robustness to changes in amino acid sequence, proteins may have evolved a native structure situated within a broad basin of low-energy conformations, a feature which could facilitate the prediction of protein structure at low resolution. PMID- 9736707 TI - DNA sequence-dependent deformability deduced from protein-DNA crystal complexes. AB - The deformability of double helical DNA is critical for its packaging in the cell, recognition by other molecules, and transient opening during biochemically important processes. Here, a complete set of sequence-dependent empirical energy functions suitable for describing such behavior is extracted from the fluctuations and correlations of structural parameters in DNA-protein crystal complexes. These elastic functions provide useful stereochemical measures of the local base step movements operative in sequence-specific recognition and protein induced deformations. In particular, the pyrimidine-purine dimers stand out as the most variable steps in the DNA-protein complexes, apparently acting as flexible "hinges" fitting the duplex to the protein surface. In addition to the angular parameters widely used to describe DNA deformations (i.e., the bend and twist angles), the translational parameters describing the displacements of base pairs along and across the helical axis are analyzed. The observed correlations of base pair bending and shearing motions are important for nonplanar folding of DNA in nucleosomes and other nucleoprotein complexes. The knowledge-based energies also offer realistic three-dimensional models for the study of long DNA polymers at the global level, incorporating structural features beyond the scope of conventional elastic rod treatments and adding a new dimension to literal analyses of genomic sequences. PMID- 9736708 TI - Tobacco mosaic virus infection induces severe morphological changes of the endoplasmic reticulum. AB - The tobacco mosaic virus (TMV) movement protein (MP) facilitates transport of virus infection between adjacent cells by modifying plasmodesmata. Previous studies suggested that the cytoskeleton and the endomembrane system are involved in this transport. We examined the effects of TMV infection on the endoplasmic reticulum (ER) in transgenic Nicotiana benthamiana that accumulate the green fluorescent protein (GFP) in the ER. Fluorescence microscopy was used to show that early in infection the ER undergoes dramatic morphological changes that include the conversion of tubular ER into large aggregates that revert to tubular ER in later stages of infection. These changes parallel MP accumulation and degradation. Furthermore, a fusion protein comprising MP fused to GFP accumulates in or on these large aggregates of ER. Expression of MP-GFP in the absence of virus infection led to the production of fluorescent aggregates of the same apparent form and size. Microsomes isolated from infected leaves contain MP. We show that the MP appears to behave as an integral ER membrane protein and is exposed on the cytosolic face of the ER. The importance of the association of MP with ER and its possible role in intracellular and intercellular spread of infection is discussed. PMID- 9736709 TI - Endoplasmic reticulum membrane localization of Rce1p and Ste24p, yeast proteases involved in carboxyl-terminal CAAX protein processing and amino-terminal a-factor cleavage. AB - Proteins terminating in the CAAX motif, for example Ras and the yeast a-factor mating pheromone, are prenylated, trimmed of their last three amino acids, and carboxyl-methylated. The enzymes that mediate these activities, collectively referred to as CAAX processing components, have been identified genetically in Saccharomyces cerevisiae. Whereas the Ram1p/Ram2p prenyltransferase is a cytosolic soluble enzyme, sequence analysis predicts that the other CAAX processing components, the Rce1p and Ste24p proteases and the Ste14p methyltransferase, contain multiple membrane spans. To determine the intracellular site(s) at which CAAX processing occurs, we have examined the localization of the CAAX proteases Rce1p and Ste24p by subcellular fractionation and indirect immunofluorescence. We find that both of these proteases are associated with the endoplasmic reticulum (ER) membrane. In addition to having a role in CAAX processing, the Ste24p protease catalyzes the first of two cleavage steps that remove the amino-terminal extension from the a-factor precursor, suggesting that the first amino-terminal processing step of a-factor maturation also occurs at the ER membrane. The ER localization of Ste24p is consistent with the presence of a carboxyl-terminal dilysine ER retrieval motif, although we find that mutation of this motif does not result in mislocalization of Ste24p. Because the ER is not the ultimate destination for a-factor or most CAAX proteins, our results imply that a mechanism must exist for the intracellular routing of CAAX proteins from the ER membrane to other cellular sites. PMID- 9736710 TI - An essential role for the phosphatidylinositol transfer protein in the scission of coatomer-coated vesicles from the trans-Golgi network. AB - We identified the phosphatidylinositol transfer protein (PITP) as being responsible for a powerful latent, nucleotide-independent, Golgi-vesiculating activity that is present in the cytosol but is only manifested as an uncontrolled activity in a cytosolic protein subfraction, in which it is separated from regulatory components that appear to normally limit its action to the scission of COPI-coated buds from trans-Golgi network membranes. A specific anti-PITP antibody that recognizes the two mammalian PITP isoforms fully inhibited the capacity of the cytosol to support normal vesicle generation as well as the uncontrolled vesiculating activity manifested by the cytosolic protein subfraction. The phosphatidylinositol- (PI) loaded form of the yeast PITP, Sec14p, but not the phosphatidylcholine- (PC) loaded form of the protein, was capable of substituting for the cytosolic subfraction in promoting the scission of coated buds from the trans-Golgi network. At higher concentration, however, Sec14p, when loaded with PI, but not with PC or phosphatidylglycerol, caused by itself an indiscriminate vesiculation of uncoated Golgi membranes that could be suppressed by PC-Sec14p, which also suppresses the uncontrolled vesiculation caused by the cytosolic subfraction. We propose that, by delivering PI to specific sites in the Golgi membrane near the necks of coated buds, PITP induces local changes in the organization of the lipid bilayer, possibly involving PI metabolites, that triggers the fusion of the ectoplasmic faces of the Golgi membrane necessary for the scission of COPI-coated vesicles. PMID- 9736712 TI - Mad2 transiently associates with an APC/p55Cdc complex during mitosis. AB - Activation of the mitotic checkpoint pathway in response to mitotic spindle damage in eukaryotic cells delays the exit from mitosis in an attempt to prevent chromosome missegregation. One component of this pathway, hsMad2, has been shown in mammalian cells to physically associate with components of a ubiquitin ligase activity (termed the anaphase promoting complex or APC) when the checkpoint is activated, thereby preventing the degradation of inhibitors of the mitotic exit machinery. In the present report, we demonstrate that the inhibitory association between Mad2 and the APC component Cdc27 also takes place transiently during the early stages of a normal mitosis and is lost before mitotic exit. We also show that Mad2 associates with the APC regulatory protein p55Cdc in mammalian cells as has been reported in yeast. In contrast, however, this complex is present only in nocodazole-arrested or early mitotic cells and is associated with the APC as a Mad2/p55Cdc/Cdc27 ternary complex. Evidence for a Mad2/Cdc27 complex that forms independent of p55Cdc also is presented. These results suggest a model for the regulation of the APC by Mad2 and may explain how the spindle assembly checkpoint apparatus controls the timing of mitosis under normal growth conditions. PMID- 9736711 TI - Transformation of hematopoietic cells by the Ski oncoprotein involves repression of retinoic acid receptor signaling. AB - The Ski oncogene has dramatic effects on the differentiation of several different cell types. It induces the differentiation of quail embryo cells into myoblasts and arrests the differentiation of chicken hematopoietic cells. The mechanism that Ski uses to carry out these disparate biological activities is unknown. However, we were struck by the similarity of these effects to those of certain members of the nuclear hormone receptor family. Both Ski and the thyroid hormone receptor-derived oncogene v-ErbA can arrest the differentiation of avian erythroblasts, and v-Ski-transformed avian multipotent progenitor cells resemble murine hematopoietic cells that express a dominant-negative form of the retinoic acid receptor, RARalpha. In this paper, we have tested the hypothesis that v-Ski and its cellular homologue c-Ski exert their effects by interfering with nuclear hormone receptor-induced transcription. We demonstrate that Ski associates with the RAR complex and can repress transcription from a retinoic acid response element. The physiological significance of this finding is demonstrated by the ability of high concentrations of a RARalpha-specific ligand to abolish v-Ski induced transformation of the multipotent progenitors. These results strongly suggest that the ability of Ski to alter cell differentiation is caused in part by the modulation of RAR signaling pathways. PMID- 9736713 TI - Coatomer, Arf1p, and nucleotide are required to bud coat protein complex I-coated vesicles from large synthetic liposomes. AB - Synthetic coat protein complex I (COPI)-coated vesicles form spontaneously from large ( approximately 300 nm in diameter), chemically defined liposomes incubated with coatomer, Arf1p, and guanosine 5'-[gamma-thio]triphosphate. Coated vesicles are 40-70 nm in diameter, approximately the size of COPI vesicles formed from native membranes. The formation of COPI-coated buds and vesicles and the binding of Arf1p to donor liposomes depends on guanosine 5'-[gamma-thio]triphosphate. In contrast to the behavior of the COPII coat, coatomer binds to liposomes containing a variety of charged or neutral phospholipids. However, the formation of COPI buds and vesicles is stimulated by acidic phospholipids. In the absence of Arf1p, coatomer binds to liposomes containing dioleoylphosphatidic acid as a sole acidic phospholipid to form large coated surfaces without forming COPI coated buds or vesicles. We conclude that Arf1p-GTP and coatomer comprise the minimum apparatus necessary to create a COPI-coated vesicle. PMID- 9736714 TI - The structural protein ODV-EC27 of Autographa californica nucleopolyhedrovirus is a multifunctional viral cyclin. AB - Two major characteristics of baculovirus infection are arrest of the host cell at G2/M phase of the cell cycle with continuing viral DNA replication. We show that Autographa californica nucleopolyhedrovirus (AcMNPV) encodes for a multifunctional cyclin that may partially explain the molecular basis of these important characteristics of AcMNPV (baculovirus) infection. Amino acids 80-110 of the viral structural protein ODV-EC27 (-EC27) demonstrate 25-30% similarity with cellular cyclins within the cyclin box. Immunoprecipitation results using antibodies to -EC27 show that -EC27 can associate with either cdc2 or cdk6 resulting in active kinase complexes that can phosphorylate histone H1 and retinoblastoma protein in vitro. The cdk6-EC27 complex also associates with proliferating cell nuclear antigen (PCNA) and we demonstrate that PCNA is a structural protein of both the budded virus and the occlusion-derived virus. These results suggest that -EC27 can function as a multifunctional cyclin: when associated with cdc2, it exhibits cyclin B-like activity; when associated with cdk6, the complex possesses cyclin D-like activity and binds PCNA. The possible roles of such a multifunctional cyclin during the life cycle of baculovirus are discussed, along with potential implications relative to the expression of functionally authentic recombinant proteins by using baculovirus-infected cells. PMID- 9736715 TI - Phosphoinositide-3-OH kinase-dependent regulation of glycogen synthase kinase 3 and protein kinase B/AKT by the integrin-linked kinase. AB - Integrin-linked kinase (ILK) is an ankyrin-repeat containing serine-threonine protein kinase capable of interacting with the cytoplasmic domains of integrin beta1, beta2, and beta3 subunits. Overexpression of ILK in epithelial cells disrupts cell-extracellular matrix as well as cell-cell interactions, suppresses suspension-induced apoptosis (also called Anoikis), and stimulates anchorage independent cell cycle progression. In addition, ILK induces nuclear translocation of beta-catenin, where the latter associates with a T cell factor/lymphocyte enhancer-binding factor 1 (TCF/LEF-1) to form an activated transcription factor. We now demonstrate that ILK activity is rapidly, but transiently, stimulated upon attachment of cells to fibronectin, as well as by insulin, in a phosphoinositide-3-OH kinase [Pi(3)K]-dependent manner. Furthermore, phosphatidylinositol(3,4,5)trisphosphate specifically stimulates the activity of ILK in vitro, and in addition, membrane targetted constitutively active Pi(3)K activates ILK in vivo. We also demonstrate here that ILK is an upstream effector of the Pi(3)K-dependent regulation of both protein kinase B (PKB/AKT) and glycogen synthase kinase 3 (GSK-3). Specifically, ILK can directly phosphorylate GSK-3 in vitro and when stably, or transiently, overexpressed in cells can inhibit GSK-3 activity, whereas the overexpression of kinase-deficient ILK enhances GSK-3 activity. In addition, kinase-active ILK can phosphorylate PKB/AKT on serine-473, whereas kinase-deficient ILK severely inhibits endogenous phosphorylation of PKB/AKT on serine-473, demonstrating that ILK is involved in agonist stimulated, Pi(3)K-dependent, PKB/AKT activation. ILK is thus a receptor proximal effector for the Pi(3)K-dependent, extracellular matrix and growth factor mediated, activation of PKB/AKT, and inhibition of GSK-3. PMID- 9736716 TI - Blocking ligand occupancy of the alphaVbeta3 integrin inhibits insulin-like growth factor I signaling in vascular smooth muscle cells. AB - Blocking alphaVbeta3 integrin occupancy results in attenuation of the cellular migration response to insulin-like growth factor I (IGF-I). To determine whether integrin antagonists alter other IGF-I-stimulated biologic actions, quiescent smooth muscle cells (SMCs) were exposed to echistatin and their ability to respond to IGF-I was determined. Echistatin (10(-7) M) inhibited IGF-I-stimulated DNA synthesis by 80%, and the protein synthesis response also was inhibited. Therefore blocking occupancy of alphaVbeta3 inhibited multiple target cell actions of IGF-I. To determine whether blocking alphaVbeta3 occupancy could alter IGF-I receptor-mediated signal transduction, the ability of IGF-I to stimulate phosphorylation of insulin receptor substrate-1 (IRS-1) was analyzed. A 10-min exposure to 100 ng/ml of IGF-I resulted in a substantial increase in phosphorylated IRS-1, and echistatin (10(-7) M) blocked the IGF-I-induced IRS-1 phosphorylation response. Echistatin also attenuated downstream signaling because the capacity of the p85 subunit of phosphatidylinositol-3 kinase (PI-3 kinase) to bind to IRS-1 was blocked. In contrast, exposure of SMCs to vitronectin (1.0 micrograms/cm2) or thrombospondin (0.25 micrograms/cm2), two known ligands for alphaVbeta3, resulted in enhancement of the IGF-I-stimulated IRS-1 response. To determine whether these effects were caused by alterations in receptor kinase activity, the IGF-I receptor was immunoprecipitated and then analyzed for phosphotyrosine. Echistatin (10(-7) M) significantly reduced IGF-I-stimulated tyrosine phosphorylation of the IGF-I receptor beta subunit. We conclude that occupancy of the alphaVbeta3 integrin is necessary for IGF-I to fully activate the kinase activity of the IGF-I receptor and phosphorylate IRS-1. Activation of the alphaVbeta3 receptor results in an interaction with the IGF-I signal transduction pathway, which modulates SMCs responsiveness to IGF-I. PMID- 9736717 TI - Vesiculation and sorting from PC12-derived endosomes in vitro. AB - Formation of small vesicles resembling synaptic vesicles can be reconstituted in vitro by incubating labeled homogenates of PC12 cells with ATP and two cytoplasmic proteins, AP3 and ARF1 [Faundez, V., Horng, J.-T. & Kelly, R. B. (1998) Cell 93, 423-432]. To determine whether AP3 was mediating budding from plasma membranes or endosomes the organelle that generated the synaptic vesicles was characterized. The budding activity was enriched in organelles that labeled at 15 degrees C, but not at 4 degrees C, that excluded a marker of plasma membranes and that contained internalized transferrin, indicating that the precursor was an endosome. Vesicles formed from the endosomal precursor in vitro excluded transferrin. We conclude that ARF-mediated vesiculation into synaptic vesicle-sized organelles uses an endosomal precursor and occurs simultaneously in vitro with sorting of synaptic vesicle proteins from other membrane protein constituents of the endosome. PMID- 9736718 TI - Interaction of HIV-1 Nef with the cellular dileucine-based sorting pathway is required for CD4 down-regulation and optimal viral infectivity. AB - The HIV-1 Nef protein is important for pathogenesis, enhances viral infectivity, and regulates the sorting of at least two cellular transmembrane proteins, CD4 and major histocompatibility complex (MHC) class I. Although several lines of evidence support the hypothesis that the Nef protein interacts directly with the cellular protein sorting machinery, the sorting signal in HIV-1 Nef has not been identified. By using a competition assay that functionally discriminates between dileucine-based and tyrosine-based sorting signals, we have categorized the motif through which Nef interacts with the sorting machinery as dileucine-based. Inspection of diverse Nef proteins from HIV-1, HIV-2, and simian immunodeficiency virus revealed a well-conserved sequence in the central region of the C-terminal, solvent-exposed loop of Nef (E/DXXXLphi) that conforms to the consensus sequence of the dileucine-based sorting motifs found in cellular transmembrane proteins. This sequence in NefNL4-3, ENTSLL, functioned as an endocytosis signal when appended to the cytoplasmic tail of a heterologous protein. The leucine residues in this motif were required for the interaction of full-length Nef with the dileucine-based sorting pathway and were required for Nef-mediated down regulation of CD4. These leucine residues were also required for optimal viral infectivity. These data indicate that a dileucine-based sorting signal in Nef is utilized to address the cellular sorting machinery. The data also suggest that an influence on the distribution of cellular transmembrane proteins may mechanistically unite two previously distinct properties of Nef: down-regulation of CD4 and enhancement of viral infectivity. PMID- 9736719 TI - Structural analysis of cloned plasma membrane proteins by freeze-fracture electron microscopy. AB - We have used freeze-fracture electron microscopy to examine the oligomeric structure and molecular asymmetry of integral plasma membrane proteins. Recombinant plasma membrane proteins were functionally expressed in Xenopus laevis oocytes, and the dimensions of their freeze-fracture particles were analyzed. To characterize the freeze-fracture particles, we compared the particle cross-sectional area of proteins with alpha-helical transmembrane domains (opsin, aquaporin 1, and a connexin) with their area obtained from existing maps calculated from two-dimensional crystals. We show that the cross-sectional area of the freeze-fracture particles corresponds to the area of the transmembrane domain of the protein, and that the protein cross-sectional area varies linearly with the number membrane-spanning helices. On average, each helix occupies 1.40 +/- 0.03 nm2. By using this information, we examined members from three classes of plasma membrane proteins: two ion channels, the cystic fibrosis transmembrane conductance regulator and connexin 50 hemi-channel; a water channel, the major intrinsic protein (the aquaporin 0); and a cotransporter, the Na+/glucose cotransporter. Our results suggest that the cystic fibrosis transmembrane conductance regulator is a dimer containing 25 +/- 2 transmembrane helices, connexin 50 is a hexamer containing 24 +/- 3 helices, the major intrinsic protein is a tetramer containing 24 +/- 3 helices, and the Na+/glucose cotransporter is an asymmetrical monomer containing 15 +/- 2 helices. PMID- 9736720 TI - Two yeast nuclear pore complex proteins involved in mRNA export form a cytoplasmically oriented subcomplex. AB - We sublocalized the yeast nucleoporin Nup82 to the cytoplasmic side of the nuclear pore complex (NPC) by immunoelectron microscopy. Moreover, by in vitro binding assays we showed that Nup82 interacts with the C-terminal region of Nup159, a yeast nucleoporin that previously was also localized to the cytoplasmic side of the NPC. Hence, the two nucleoporins, Nup82 and Nup159, form a cytoplasmically oriented subcomplex that is likely to be part of the fibers emanating from the cytoplasmic ring of the NPC. Overexpression of Rss1/Gle1, a putative nucleoporin and/or mRNA transport factor, was shown previously to partially rescue depletion of Nup159. We show here that overexpression of Rss1/Gle1 also partially rescued depletion of Nup82. Depletion of either Nup82, Nup159, or Rss1/Gle1 was shown previously to inhibit mRNA export. As was reported previously for depletion of Nup159 or of Rss1/Gle1, we show here that depletion of Nup82 has no detectable effect on classical nuclear localization sequence mediated nuclear import. In summary, the nucleoporins Nup159 and Nup82 form a cytoplasmically oriented subcomplex of the NPC that is likely associated with Rss1/Gle1; this complex is essential for RNA export, but not for classical nuclear localization sequence-mediated nuclear protein import. PMID- 9736721 TI - Very late DNA replication in the human cell cycle. AB - G2 was defined originally as the temporal gap between the termination of DNA replication and the beginning of mitosis. In human cells, the G2 period was estimated to be 3-4 h. However, the absence of replicative DNA synthesis during this period designated G2 has never been shown conclusively. In this report, we show that, at some autosomal and X linked loci, programmed DNA replication continues within 90 min of mitosis. Furthermore, the major accumulation of cyclin B1, a cell-cycle marker that is usually ascribed to G2, overlaps extensively with very late DNA replication. We conclude that the G2 period is much shorter than previously thought and may, in some cells, be nonexistent. PMID- 9736723 TI - Identification and characterization of LMO4, an LMO gene with a novel pattern of expression during embryogenesis. AB - LMO4 is a novel member of the LIM-only (LMO) subfamily of LIM domain-containing transcription factors. LMO1, LMO2, and LMO4 have distinct expression patterns in adult tissue, and we demonstrate that nuclear retention of LMO proteins is enhanced by the nuclear LIM interactor (NLI). In situ hybridization to early mouse embryos of 8-14.5 days revealed a complex pattern of LMO4 expression spatially overlapping with NLI and LHX genes. LMO4 expression in somite is repressed in mice mutant for the segment polarity gene Mesp2 and expanded in Splotch mutants. During jaw and limb outgrowth, LMO4 and LMO2 expression define mesenchyme that is uncommitted to regional fates. Although both LMO2 and LMO4 are activated in thymic blast cells, only LMO4 is expressed in mature T cells. Mesenchymal and thymic blast cell expression patterns of LMO4 and LMO2 are consistent with the suggestion that LMO genes inhibit differentiation. PMID- 9736722 TI - Cloning of Mix-related homeodomain proteins using fast retrieval of gel shift activities, (FROGS), a technique for the isolation of DNA-binding proteins. AB - We have developed a technique, fast retrieval of gel shift activities (FROGS), that allows for the rapid isolation of proteins that interact with DNA. Using this technique, we have isolated two proteins that are structurally similar to Mix.1, a PAX class homeodomain protein with ventralizing activity in Xenopus. The Mix family of proteins are expressed during late blastula and gastrula stages of Xenopus development. During gastrulation, these genes are expressed at high levels in distinct, yet overlapping regions in mesoderm and endoderm. The members of the Mix family heterodimerize with each other and overexpression of each results in severe axial abnormalities. Mix.3 and Mix.4 can directly induce primitive ectoderm to become endoderm whereas Mix.1 cannot. Injection of Mix.3 or Mix.4 RNA in the whole embryo results in extensive ectopic endodermin mRNA expression. The expression of the Mix family homeoproteins is differentially regulated by activin, Vg1, BMP-4, and fibroblast growth factor, supporting a model in which the Mix homeoproteins are downstream effectors of growth factor signaling during endoderm and ventral mesoderm formation. PMID- 9736725 TI - Structure elucidation and chemical synthesis of stigmolone, a novel type of prokaryotic pheromone. AB - Approximately 2 micromol of a novel prokaryotic pheromone, involved in starvation induced aggregation and formation of fruiting bodies by the myxobacterium Stigmatella aurantiaca, were isolated by a large-scale elution procedure. The pheromone was purified by HPLC, and high-resolution MS, IR, 1H-NMR, and 13C-NMR were used to identify the active substance as the hydroxy ketone 2,5, 8-trimethyl 8-hydroxy-nonan-4-one, which has been named stigmolone. The analysis was complicated by a solvent-dependent equilibrium between stigmolone and the cyclic enol-ether 3,4-dihydro-2,2, 5-trimethyl-6-(2-methylpropyl)-2H-pyran formed by intramolecular nucleophilic attack of the 8-OH group at the ketone C4 followed by loss of H2O. Both compounds were synthesized chemically, and their structures were confirmed by NMR analysis. Natural and synthetic stigmolone have the same biological activity at ca. 1 nM concentration. PMID- 9736724 TI - Intercellular signaling in Stigmatella aurantiaca: purification and characterization of stigmolone, a myxobacterial pheromone. AB - The myxobacterium Stigmatella aurantiaca passes through a life cycle that involves formation of a multicellular fruiting body as the most complex stage. An early step in this differentiation process depends on a signal factor secreted by the cells when nutrients become limited. The formation of a fruiting body from a small cell population can be accelerated by addition of this secreted material. The bioactive compound was found to be steam volatile. It was purified to homogeneity by steam distillation followed by reversed-phase and normal-phase HPLC. The pheromone was named stigmolone, in accordance with the structure 2,5, 8 trimethyl-8-hydroxy-nonan-4-one, as determined by NMR and mass spectrometry. Stigmolone represents a structurally unique and highly bioactive prokaryotic pheromone that is effective in the bioassay at 1 nM concentration. PMID- 9736726 TI - Essential role of mitochondrially encoded large rRNA for germ-line formation in Drosophila embryos. AB - In Drosophila, pole cells, the progenitors of the germ line, are induced by the factors localized in the posterior pole region of oocytes and cleavage embryos, or germ plasm. Polar granules in germ plasm are electron-dense structures and have been proposed to contain factors essential for pole cell formation. Mitochondrially encoded large ribosomal RNA (mtlrRNA) has been identified as a component of polar granules. We previously have shown that mtlrRNA is able to rescue embryos that fail to form pole cells as a result of UV irradiation. However, there is a possibility that the function of mtlrRNA is limited to UV irradiated embryos, and the question of whether mtlrRNA is required for the normal pathway leading to pole cell formation remains unanswered. In this study, we report that the reduction of mtlrRNA in germ plasm by injecting anti-mtlrRNA ribozymes into cleavage embryos leads to their inability to form pole cells. Other components of germ plasm, namely oskar mRNA, germ cell-less mRNA, and Vasa and Tudor proteins appear to be unaffected in these ribozyme-injected embryos. These results support an essential role for mtlrRNA in pole cell formation. We propose that mitochondrially encoded molecules participate in a key event in early cell-type specification. PMID- 9736727 TI - Diversity components of impending primate extinctions. AB - Many extant species are at risk to go extinct. This impending loss of species is likely to cause changes in future ecosystem functions. Ecological components of diversity, such as dietary or habitat specializations, can be used to estimate the impact of extinctions on ecosystem functions. As an approach to estimate the impact of future extinctions, we tested interdependency between ecological and taxonomic change based on current predictions of extinction rates in primates. We analyzed the ecological characteristics of extant primate faunas having species in various categories of endangerment of extinction and forecasted the future primate faunas as if they were paleontological faunas. Predicting future faunas combines the wealth of ecological information on living primates with large, fossil record-like changes in diversity. Predicted extinction patterns of living primates in Africa, Asia, Madagascar, and South America show that changes in ecology differ among the regions in ways that are not reducible to taxonomic measures. The ecological effects of primate extinctions are initially least severe in South America and larger in Asia and Africa. Disproportionately larger ecological changes are projected for Madagascar. The use of taxonomy as a proxy for ecology can mislead when estimating competence of future primate ecosystems. PMID- 9736728 TI - Determining and dating recent rodent speciation events by using L1 (LINE-1) retrotransposons. AB - Phylogenies based on the inheritance of shared derived characters will be ambiguous when the shared characters are not the result of common ancestry. Such characters are called homoplasies. Phylogenetic analysis also can be problematic if the characters have not changed sufficiently, as might be the case for rapid or recent speciations. The latter are of particular interest because evolutionary processes may be more accessible the more recent the speciation. The repeated DNA subfamilies generated by the mammalian L1 (LINE-1) retrotransposon are apparently homoplasy-free phylogenetic characters. L1 retrotransposons are transmitted only by inheritance and rapidly generate novel variants that produce distinct subfamilies of mostly defective copies, which then "age" as they diverge. Here we show that the L1 character can both resolve and date recent speciation events within the large group of very closely related rats known as Rattus sensu stricto. This lineage arose 5-6 million years ago (Mya) and subsequently underwent two episodes of speciation: an intense one, approximately 2.7 Mya, produced at least five lineages in <0.3 My; a second began approximately 1.2 Mya and may still be continuing. PMID- 9736729 TI - The adaptive nature of the human neurocognitive architecture: an alternative model. AB - The model of the human neurocognitive architecture proposed by evolutionary psychologists is based on the presumption that the demands of hunter-gatherer life generated a vast array of cognitive adaptations. Here we present an alternative model. We argue that the problems inherent in the biological markets of ancestral hominids and their mammalian predecessors would have required an adaptively flexible, on-line information-processing system, and would have driven the evolution of a functionally plastic neural substrate, the neocortex, rather than a confederation of evolutionarily prespecified social cognitive adaptations. In alignment with recent neuroscientific evidence, we suggest that human cognitive processes result from the activation of constructed cortical representational networks, which reflect probabilistic relationships between sensory inputs, behavioral responses, and adaptive outcomes. The developmental construction and experiential modification of these networks are mediated by subcortical circuitries that are responsive to the life history regulatory system. As a consequence, these networks are intrinsically adaptively constrained. The theoretical and research implications of this alternative evolutionary model are discussed. PMID- 9736730 TI - Genetic code origins: tRNAs older than their synthetases? AB - We present a phylogenetic analysis to determine whether a given tRNA molecule was established in evolution before its cognate aminoacyl-tRNA synthetase. The earlier appearance of tRNA versus their metabolically related enzymes is a prediction of the RNA world theory, but the available synthetase and tRNA sequences previously had not allowed a formal comparison of their relative time of appearance. Using data recently obtained from the emerging genome projects, our analysis points to the extant forms of lysyl-tRNA synthetase being preceded in evolution by the establishment of the identity of lysine tRNA. PMID- 9736731 TI - Aspirin suppresses the mutator phenotype associated with hereditary nonpolyposis colorectal cancer by genetic selection. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) are well-known cancer preventives, which have been largely attributed to their antiproliferative and apoptosis inducing activities. In this study, we show that microsatellite instability (MSI) in colorectal cancer cells deficient for a subset of the human mismatch repair (MMR) genes (hMLH1, hMSH2, and hMSH6), is markedly reduced during exposure to aspirin or sulindac [or Clinoril, which is chemically related to indomethacin (Indocin)]. This effect was reversible, time and concentration dependent, and appeared independent of proliferation rate and cyclooxygenase function. In contrast, the MSI phenotype of a hPMS2-deficient endometrial cancer cell line was unaffected by aspirin/sulindac. We show that the MSI reduction in the susceptible MMR-deficient cells was confined to nonapoptotic cells, whereas apoptotic cells remained unstable and were eliminated from the growing population. These results suggest that aspirin/sulindac induces a genetic selection for microsatellite stability in a subset of MMR-deficient cells and may provide an effective prophylactic therapy for hereditary nonpolyposis colorectal cancer kindreds where alteration of the hMSH2 and hMLH1 genes are associated with the majority of cancer susceptibility cases. PMID- 9736732 TI - The expression of the KAI1 gene, a tumor metastasis suppressor, is directly activated by p53. AB - KAI1 is a tumor metastasis suppressor gene that is capable of inhibiting the metastatic process in animals. The expression of the KAI1 gene also is found to be down-regulated during the tumor progression of prostate, breast, lung, bladder, and pancreatic cancers in humans, and this down-regulation appears to be at or posttranscription level. We have found that the tumor suppressor gene p53 can directly activate the KAI1 gene by interacting with the 5' upstream region. The p53 responding region is located at approximately 860 bases upstream of the transcriptional initiation site, and it contains a typical tandem repeat of the p53 consensus-binding sequence. A gel-shift mobility analysis showed that this sequence indeed had the ability to bind to the purified p53 protein. Mutations of this sequence abolished the responsiveness to p53 and also the binding ability to the p53 protein. Furthermore, immunohistochemical analysis of 177 samples of human prostate tumors revealed that the expression of the KAI1 gene was correlated strongly to that of the p53 gene and that the loss of these two markers resulted in poor survivals of patients. Our data indicate a direct relationship between p53 and KAI1 genes and suggest that the loss of p53 function, which is commonly observed in many types of cancer, leads to the down regulation of the KAI1 gene, which may result in the progression of metastasis. PMID- 9736733 TI - A dysfunctional desmin mutation in a patient with severe generalized myopathy. AB - Mice lacking desmin produce muscle fibers with Z disks and normal sarcomeric organization. However, the muscles are mechanically fragile and degenerate upon repeated contractions. We report here a human patient with severe generalized myopathy and aberrant intrasarcoplasmic accumulation of desmin intermediate filaments. Muscle tissue from this patient lacks the wild-type desmin allele and has a desmin gene mutation encoding a 7-aa deletion within the coiled-coil segment of the protein. We show that recombinant desmin harboring this deletion cannot form proper desmin intermediate filament networks in cultured cells, nor is it able to assemble into 10-nm filaments in vitro. These findings provide direct evidence that a mutation in desmin can cause human myopathies. PMID- 9736734 TI - Construction and validation of yeast artificial chromosome contig maps by RecA assisted restriction endonuclease cleavage. AB - RecA-assisted restriction endonuclease (RARE) cleavage is an "Achilles' heel" approach to restriction mapping whereby a RecA-protein-oligodeoxynucleotide complex protects an individual restriction site from methylation, thus limiting subsequent digestion to a single, predetermined site. We have used RARE cleavage to cut yeast artificial chromosomes (YACs) at specific EcoRI sites located within or adjacent to sequence-tagged sites (STSs). Each cleavage reaction produces two YAC fragments whose sizes are a direct measure of the position of the STS in the YAC. In this fashion, we have positioned 45 STSs within a contig of 19 independent YACs and constructed a detailed RARE-cleavage map that represents 8.4 Mbp of human chromosome 6p21.3-22. By comparing maps of overlapping YACs, we were able to detect seven internal deletions that ranged from approximately 75 kbp to approximately 1 Mbp in size. Thirteen pairs of EcoRI sites were targeted for double RARE cleavage in uncloned total human DNA. The excised fragments, up to 2 Mbp in size, were resolved by pulsed-field gel electrophoresis and were detected by hybridization. In general, the genomic RARE-cleavage results support the YAC based map. In one case, the distance in uncloned DNA between the two terminal EcoRI sites of a YAC insert was approximately 1 Mbp larger than the YAC itself, indicating a major deletion. The general concept of RARE-cleavage mapping as well as its applications and limitations are discussed. PMID- 9736735 TI - Human CUL-1 associates with the SKP1/SKP2 complex and regulates p21(CIP1/WAF1) and cyclin D proteins. AB - Deregulation of cell proliferation is a hallmark of cancer. In many transformed cells, the cyclin A/CDK2 complex that contains S-phase kinase associated proteins 1 and 2 (SKP1 and SKP2) is highly induced. To determine the roles of this complex in the cell cycle regulation and transformation, we have examined the composition of this complex. We report here that this complex contained an additional protein, human CUL-1, a member of the cullin/CDC53 family. The identification of CUL-1 as a member of the complex raises the possibility that the p19(SKP1)/p45(SKP2)/CUL-1 complex may function as the yeast SKP1-CDC53-F-box (SCF) protein complex that acts as a ubiquitin E3 ligase to regulate the G1/S transition. In mammalian cells, cyclin D, p21(CIP1/WAF1), and p27(KIP1) are short lived proteins that are controlled by ubiquitin-dependent proteolysis. To determine the potential in vivo targets of the p19(SKP1)/p45(SKP2)/CUL-1 complex, we have used the specific antisense oligodeoxynucleotides against either SKP1, SKP2, or CUL-1 RNA to inhibit their expression. Treatment of cells with these oligonucleotides caused the selective accumulation of p21 and cyclin D proteins. The protein level of p27 was not affected. These data suggest that the human p19(SKP1)/p45(SKP2)/CUL-1 complex is likely to function as an E3 ligase to selectively target cyclin D and p21 for the ubiquitin-dependent protein degradation. Aberrant expression of human p19(SKP1)/p45(SKP2)/CUL-1 complex thus may contribute to tumorigenesis by regulating the protein levels of G1 cell cycle regulators. PMID- 9736736 TI - The src homology 2-containing inositol phosphatase (SHIP) is the gatekeeper of mast cell degranulation. AB - To clarify the role that the src homology 2-containing inositol phosphatase (SHIP) plays in mast cell degranulation, the gene for SHIP was disrupted by homologous recombination in embryonic stem cells. Bone-marrow-derived mast cells from SHIP+/+, +/-, and -/- F2 littermates were compared. SHIP-/- mast cells were found to be far more prone to degranulation, after the crosslinking of IgE preloaded cells, than SHIP+/- or +/+ cells. Intriguingly, IgE alone also stimulated massive degranulation in SHIP-/- but not in +/+ mast cells. This degranulation with IgE alone, which may be due to low levels of IgE aggregates, correlated with a higher and more sustained intracellular calcium level than that observed with SHIP+/+ cells and was dependent upon the entry of extracellular calcium. Immunoprecipitation studies revealed that the addition of IgE alone to normal mast cells stimulates multiple cascades, which are prevented from progressing to degranulation by SHIP. PI 3-kinase inhibitor studies suggested that IgE-induced activation of PI 3-kinase is upstream of the entry of extracellular calcium and that SHIP restricts this entry by hydrolyzing phosphatidylinositol 3,4, 5-trisphosphate. These results show the critical role that SHIP plays in setting the threshold for degranulation and that SHIP directly modulates a "positive-acting" receptor. PMID- 9736737 TI - Attractin (DPPT-L), a member of the CUB family of cell adhesion and guidance proteins, is secreted by activated human T lymphocytes and modulates immune cell interactions. AB - Attractin is a normal human serum glycoprotein of 175 kDa that is rapidly expressed on activated T cells and released extracellularly after 48-72 hr. We have cloned attractin and find that, as in its natural serum form, it mediates the spreading of monocytes that become the focus for the clustering of nonproliferating T lymphocytes. There are two mRNA species with hematopoietic tissue-specific expression that code for a 134-kDa protein with a putative serine protease catalytic serine, four EGF-like motifs, a CUB domain, a C type lectin domain, and a domain homologous with the ligand-binding region of the common gamma cytokine chain. Except for the latter two domains, the overall structure shares high homology with the Caenorhabditis elegans F33C8.1 protein, suggesting that attractin has evolved new domains and functions in parallel with the development of cell-mediated immunity. PMID- 9736738 TI - Differential display of peptides induced during the immune response of Drosophila: a matrix-assisted laser desorption ionization time-of-flight mass spectrometry study. AB - We have developed an approach based on a differential mass spectrometric analysis to detect molecules induced during the immune response of Drosophila, regardless of their biological activities. For this, we have applied directly matrix assisted laser desorption/ionization MS to hemolymph samples from individual flies before and after an immune challenge. This method provided precise information on the molecular masses of immune-induced molecules and allowed the detection, in the molecular range of 1.5-11 kDa, of 24 Drosophila immune-induced molecules (DIMs). These molecules are all peptides, and four correspond to already characterized antimicrobial peptides. We have further analyzed the induction of the various peptides by immune challenge in wild-type flies and in mutants with a compromised antimicrobial response. We also describe a methodology combining matrix-assisted laser desorption ionization time-of-flight MS, HPLC, and Edman degradation, which yielded the peptide sequence of three of the DIMs. Finally, molecular cloning and Northern blot analyses revealed that one of the DIMs is produced as a prepropeptide and is inducible on a bacterial challenge. PMID- 9736739 TI - Regulatory dysfunction of the interleukin-2 receptor during HIV infection and the impact of triple combination therapy. AB - The interleukin-2 (IL-2)/IL-2 receptor (IL-2R) system is the main regulatory determinant of T cell reactivity. Although it is well known that IL-2 secretion is impaired during HIV infection, up to now IL-2R expression has not been extensively studied in HIV-infected patients despite the use of IL-2 in clinical therapy trials. We show here that IL-2R expression in HIV patients with high viral load (group 1 in the study) is greatly enhanced on B lymphocytes, CD8 T lymphocytes, and monocytes, but not on CD4 T lymphocytes, compared with noninfected individuals. Paradoxically, this modified IL-2R expression does not lead to increased IL-2 responsiveness, except for B lymphocytes. In patients receiving triple combination therapy (TCT, two reverse transcriptase inhibitors and one protease inhibitor) that has triggered a drastic reduction in plasma viral load and an increase in CD4 counts (group 2 patients), IL-2R expression is significantly lower than in group 1 patients. Moreover, cells involved in cellular immunity and CD4 T lymphocytes have the capacity to respond to IL-2 after TCT. These results allow us to anticipate a beneficial role of IL-2 immunotherapy in combination with TCT. PMID- 9736740 TI - Transgenic mice for interleukin 3 develop motor neuron degeneration associated with autoimmune reaction against spinal cord motor neurons. AB - Interleukin 3 (IL-3) stimulates the proliferation and differentiation of various haematopoietic progenitor cells. Recently, IL-3 and other cytokines were reported to exert a neurotrophic activity and to be associated with neurological disorders, suggesting their complex role in the central nervous system. We now show that overexpression of IL-3 in transgenic mice causes a motor neuron disease with several features of amyotrophic lateral sclerosis and progressive muscular atrophy. These animals exhibit hind limb paralysis at 7 months of age, associated with dendritic and axonal degeneration, loss of motor neurons in the spinal cord, and autoimmune reaction against these cells. We examined the effect of IL-3 on embryonic motor neurons survival in mixed spinal cord cultures. Our results suggest that motor neuronal degeneration is not directly triggered by the high level of expression of IL-3. PMID- 9736741 TI - HIV type I envelope determinants for use of the CCR2b, CCR3, STRL33, and APJ coreceptors. AB - The envelope (Env) proteins of primate lentiviruses interact sequentially with CD4 and a coreceptor to infect cells. Changes in coreceptor use strongly influence viral tropism and pathogenesis. We followed the evolution of coreceptor use in pig-tailed macaques that developed severe CD4 T-cell loss during the derivation of a pathogenic simian HIV (SHIV) that contained the tat, rev, vpu, and env genes of the HXBc2 strain of HIV-1 in a genetic background of SIVmac239. The Env from the parental virus as well as one derived from the first macaque to develop AIDS exclusively used CXCR4 as a coreceptor, indicating that CXCR4 can function as a coreceptor in macaques even though it is rarely used by simian immunodeficiency viruses. One Env (Pnb5), obtained from a macrophage-tropic virus isolated from the cerebral spinal fluid, did not use CCR5 or CXCR4. Instead, it used CCR2b and to a lesser extent CCR3, STRL33, and APJ to infect cells. Chimeras between Pnb5 and the parental X4 Env indicated that the V3 loop is the major determinant of CXCR4 use, with other regions of Env influencing the efficiency with which this coreceptor was used. In contrast, the Pnb5 V1/2 and V3 regions in combination were both necessary and sufficient to confer full use of CCR2b, CCR3, STRL33, and APJ to the parental X4 Env protein. These results are consistent with a single, conserved binding site in Env that interacts with multiple coreceptors in conjunction with the V1/2 and V3 loops, and suggest that the V1/2 region plays a more important role in governing the use of CCR2b, CCR3, STRL33, and APJ than for CXCR4. PMID- 9736742 TI - Allelic association under map error and recombinational heterogeneity: a tale of two sites. AB - Recombination acts on the genetic map, not on the physical map. On the other hand, the physical map is usually more accurate. Choice of the genetic or physical map for positional cloning by allelic association depends on the goodness of fit of data to each map under an established model. Huntington disease illustrates the usual case in which the greater reliability of physical data outweighs recombinational heterogeneity. Hemochromatosis represents an exceptional case in which unrecognized recombinational heterogeneity retarded positional cloning for a decade. The Malecot model performs well for major genes, but no approach assuming either equilibrium or disequilibrium has been validated for oligogenes contributing to common disease. In this case of greatest interest, the power of allelic association relative to linkage is less clear than for major genes. PMID- 9736743 TI - Conditional lineage ablation to model human diseases. AB - Cell loss contributes to the pathogenesis of many inherited and acquired human diseases. We have developed a system to conditionally ablate cells of any lineage and developmental stage in the mouse by regulated expression of the diphtheria toxin A (DTA) gene by using tetracycline-responsive promoters. As an example of this approach, we targeted expression of DTA to the hearts of adult mice to model structural abnormalities commonly observed in human cardiomyopathies. Induction of DTA expression resulted in cell loss, fibrosis, and chamber dilatation. As in many human cardiomyopathies, transgenic mice developed spontaneous arrhythmias in vivo, and programmed electrical stimulation of isolated-perfused transgenic hearts demonstrated a strikingly high incidence of spontaneous and inducible ventricular tachycardia. Affected mice showed marked perturbations of cardiac gap junction channel expression and localization, including a subset with disorganized epicardial activation patterns as revealed by optical action potential mapping. These studies provide important insights into mechanisms of arrhythmogenesis and suggest that conditional lineage ablation may have wide applicability for studies of disease pathogenesis. PMID- 9736746 TI - Tests and estimates of allelic association in complex inheritance. AB - Family-based procedures such as the transmission disequilibrium test (TDT) were motivated by concern that sample-based methods to map disease genes by allelic association are not robust to population stratification, migration, and admixture. Other factors to consider in designing a study of allelic association are specification of gene action in a weakly parametric model, efficiency, diagnostic reliability for hypernormal individuals, interest in linkage and imprinting, and sibship composition. Family-based samples lend themselves to the TDT despite its inefficiency compared with cases and unrelated normal controls. The TDT has an efficiency of 1/2 for parent-offspring pairs and 2/3 for father mother-child trios. Against cases and hypernormal controls, the efficiency is only 1/6 on the null hypothesis. Although dependent on marker gene frequency and other factors, efficiency for hypernormal controls is always greater than for random controls. Efficiency of the TDT is increased in multiplex families and by inclusion of normal sibs, approaching a case-control design with normal but not hypernormal controls. Isolated cases favor unrelated controls, and only in exceptional populations would avoidance of stratification justify a family-based design to map disease genes by allelic association. PMID- 9736744 TI - Cellular immune response to adenoviral vector infected cells does not require de novo viral gene expression: implications for gene therapy. AB - Replication-defective adenoviral (RDAd) vectors can be generated at high titers and infect both dividing and nondividing cells. Long term expression in the transduced tissue, however, has been a problem because of the cellular immune responses against the infected cells. We demonstrate that mice injected with RDAd vectors containing mouse leptin gene reduce food intake and lose weight for only 7 to 10 days. Splenocytes obtained from infected mice are able to lyse target cells infected with RDAd vectors. Surprisingly, target cells infected with psoralen-treated, UV-crosslinked, biologically inactive RDAd also were lysed efficiently by the effector cells. Furthermore, splenocytes obtained from mice injected with inactive RDAd vectors efficiently lysed target cells infected with RDAd vectors. Whether RDAd vectors were injected i.m. or i.v. or through an i.p. route, the extent of lysis was similar. We propose that cells infected with RDAd vectors present antigens for recognition by class 1 major histocompatibility complex-restricted cytotoxic T lymphocytes by a mechanism that does not require viral replication or de novo protein synthesis. These results should prompt reevaluation of the use of RDAd vectors for gene therapy when long-term expression is required. PMID- 9736745 TI - Macrophage migration inhibitory factor is a critical mediator of the activation of immune cells by exotoxins of Gram-positive bacteria. AB - Discovered in the early 1960s as a T cell cytokine, the protein mediator known as macrophage migration inhibitory factor (MIF) has been found recently to be a pituitary peptide released during the physiological stress response, a proinflammatory macrophage cytokine secreted after LPS stimulation, and a T cell product expressed as part of the antigen-dependent activation response. We report herein that MIF also plays a critical role in the innate host response to staphylococcal and streptococcal exotoxins. In RAW 264.7 or elicited mouse peritoneal macrophages, peak MIF secretion was induced by concentrations of the staphylococcal toxic shock syndrome (TSS) toxin 1 (TSST-1) and the streptococcal pyrogenic exotoxin A as low as 10 pg/ml. Moreover, dose-response studies of splenocyte cytokine production showed that lower concentrations of TSST-1 (10 pg/ml) were needed to release MIF than to induce interleukin 2 or interferon gamma secretion (1 ng/ml). We also studied the effect of neutralizing anti-MIF antibodies on TSST-1-induced lymphocyte proliferation and lethal toxic shock. Pretreatment of C57BL/6 mice with anti-MIF antibody 2 hr before TSST-1 injection prevented spleen enlargement and reduced by 50% the proliferation of splenocytes measured ex vivo. In a lethal mouse model of TSST-1-induced shock, anti-MIF antibody increased survival from 8% to 54% (P < 0.0001). These studies indicate that Gram-positive exotoxins are extremely potent inducers of MIF secretion and establish a critical role for MIF and the macrophage in the pathogenesis of the TSSs and in the innate immune response. PMID- 9736747 TI - Presence of a gene encoding choline sulfatase in Sinorhizobium meliloti bet operon: choline-O-sulfate is metabolized into glycine betaine. AB - Glycine betaine is a potent osmoprotectant accumulated by Sinorhizobium meliloti to cope with osmotic stress. The biosynthesis of glycine betaine from choline is encoded by an operon of four genes, betICBA, as determined by sequence and mutant analysis. The betI and betC genes are separated by an intergenic region containing a 130-bp mosaic element that also is present between the betB and betA genes. In addition to the genes encoding a presumed regulatory protein (betI), the betaine aldehyde dehydrogenase (betB), and the choline dehydrogenase (betA) enzymes also found in Escherichia coli, a new gene (betC) was identified as encoding a choline sulfatase catalyzing the conversion of choline-O-sulfate and, at a lower rate, phosphorylcholine, into choline. Choline sulfatase activity was absent from betC but not from betB mutants and was shown to be induced indifferently by choline or choline-O-sulfate as were the other enzymes of the pathway. Unlike what has been shown in other bacteria and plants, choline-O sulfate is not used as an osmoprotectant per se in S. meliloti, but is metabolized into glycine betaine. S. meliloti also can use this compound as the sole carbon, nitrogen, and sulfur source for growth and that depends on a functional bet locus. In conclusion, choline-O-sulfate and phosphorylcholine, which are found in higher plants and fungi, appear to be substrates for glycine betaine biosynthesis in S. meliloti. PMID- 9736748 TI - Molecular markers for cell types of the inner ear and candidate genes for hearing disorders. AB - To identify genes expressed in the vertebrate inner ear, we have established an assay that allows rapid analysis of the differential expression pattern of mRNAs derived from an auditory epithelium-specific cDNA library. We performed subtractive hybridization to create an enriched probe, which then was used to screen the cDNA library. After digoxigenin-labeled antisense cRNAs had been transcribed from hybridization-positive clones, we conducted in situ hybridization on slides bearing cryosections of late embryonic chicken heads, bodies, and cochleae. One hundred and twenty of the 196 clones analyzed encode 12 proteins whose mRNAs are specifically or highly expressed in the chicken's inner ear; the remainder encode proteins that occur more widely. We identified proteins that have been described previously as expressed in the inner ear, such as beta tectorin, calbindin, and type II collagen. A second group of proteins abundant in the inner ear includes five additional types of collagens. A third group, including Coch-5B2 and an ear-specific connexin, comprises proteins whose human equivalents are candidates to account for hearing disorders. This group also includes proteins expressed in two unique cell types of the inner ear, homogene cells and cells of the tegmentum vasculosum. PMID- 9736749 TI - Hippocampal morphometry in schizophrenia by high dimensional brain mapping. AB - Theories of the pathophysiology of schizophrenia have implicated the hippocampus, but controversy remains regarding hippocampal abnormalities in patients with schizophrenia. In vivo studies of hippocampal anatomy using high resolution magnetic resonance scanning and manual methods for volumetric measurement have yielded inconclusive results, perhaps because of the normal variability in hippocampal volume and the error involved in manual measurement techniques. To resolve this controversy, high dimensional transformations of a computerized brain template were used to compare hippocampal volumes and shape characteristics in 15 matched pairs of schizophrenia and control subjects. The transformations were derived from principles of general pattern matching and were constrained according to the physical properties of fluids. The analysis and comparison of hippocampal shapes based on these transformations were far superior to the comparison of hippocampal volumes or other global indices of hippocampal anatomy in showing a statistically significant difference between the two groups. In the schizophrenia subjects, hippocampal shape deformations were found to be localized to subregions of the structure that send projections to prefrontal cortex. The results of this study demonstrate that abnormalities of hippocampal anatomy occur in schizophrenia and support current hypotheses that schizophrenia involves a disturbance of hippocampal-prefrontal connections. These results also show that comparisons of neuroanatomical shapes can be more informative than volume comparisons for identifying individuals with neuropsychiatric diseases, such as schizophrenia. PMID- 9736750 TI - Brain-derived neurotrophic factor mediates estradiol-induced dendritic spine formation in hippocampal neurons. AB - Dendritic spines are of major importance in information processing and memory formation in central neurons. Estradiol has been shown to induce an increase of dendritic spine density on hippocampal neurons in vivo and in vitro. The neurotrophin brain-derived neurotrophic factor (BDNF) recently has been implicated in neuronal maturation, plasticity, and regulation of GABAergic interneurons. We now demonstrate that estradiol down-regulates BDNF in cultured hippocampal neurons to 40% of control values within 24 hr of exposure. This, in turn, decreases inhibition and increases excitatory tone in pyramidal neurons, leading to a 2-fold increase in dendritic spine density. Exogenous BDNF blocks the effects of estradiol on spine formation, and BDNF depletion with a selective antisense oligonucleotide mimics the effects of estradiol. Addition of BDNF antibodies also increases spine density, and diazepam, which facilitates GABAergic neurotransmission, blocks estradiol-induced spine formation. These observations demonstrate a functional link between estradiol, BDNF as a potent regulator of GABAergic interneurons, and activity-dependent formation of dendritic spines in hippocampal neurons. PMID- 9736751 TI - Role of the Doc2 alpha-Munc13-1 interaction in the neurotransmitter release process. AB - Doc2alpha and Munc13-1 proteins are highly concentrated on synaptic vesicles and the presynaptic plasma membrane, respectively, and have been implicated in Ca2+ dependent neurotransmitter release. Doc2alpha interacts with Munc13-1 through the N-terminal region of Doc2alpha (the Mid domain; amino acid residues 13-37). Here we examine whether the interaction between Doc2alpha and Munc13-1 is required for Ca2+-dependent neurotransmitter release from intact neuron. A synthetic Mid peptide (the Mid peptide), but not a control mutated Mid peptide or a scrambled Mid peptide, inhibited the interaction between Doc2alpha and Munc13-1 in vitro. Introduction of the Mid peptide into presynaptic neurons of cholinergic synapses, formed between rat superior cervical ganglion neurons, reversibly inhibited synaptic transmission evoked by action potentials. In contrast, the control peptides did not inhibit synaptic transmission. This inhibitory effect depended on the presynaptic activity and was affected by extracellular Ca2+ concentrations. The onset of the Mid peptide effect was shortened when the neuron was stimulated at a higher frequency, and the inhibition was more potent at 1 mM Ca2+ than at 5.1 mM Ca2+. These results suggest that the Doc2alpha-Munc13-1 interaction plays a role in a step before the final fusion step of synaptic vesicles with the presynaptic plasma membrane in the evoked neurotransmitter release process. PMID- 9736752 TI - gas7: A gene expressed preferentially in growth-arrested fibroblasts and terminally differentiated Purkinje neurons affects neurite formation. AB - Growth arrest-specific (gas) genes are expressed preferentially in cells that enter a quiescent state. gas7, which we identified in serum-starved murine fibroblasts, is reported here to be expressed in vivo selectively in neuronal cells of the mature cerebral cortex, hippocampus, and cerebellum. gas7 transcripts encode a 48-kDa protein containing a structural domain that resembles sequences of OCT2, a POU transcription factor implicated in neuronal development, and synapsins, which have a role in modulating neurotransmitter release. Using in situ hybridization and immunocytochemical analysis, we show that GAS7 expression occurs prominently in cerebellar Purkinje cells and that inhibition of production in terminally differentiating cultures of embryonic murine cerebellum impedes neurite outgrowth from maturing Purkinje cells. Conversely, GAS7 overexpression in undifferentiated neuroblastoma cell cultures dramatically promotes neurite like outgrowth. Collectively, our results provide evidence for an association between expression of this gas gene and neuronal development. PMID- 9736754 TI - Immediate and long-term effects of neonatal MK-801 treatment on nonspatial learning. AB - These experiments observed the immediate and long-term effects of neonatal treatment with MK-801 on patterned single alternation (PSA), a form of nonspatial, memory-based learning. Rat pups were injected daily on postnatal days (PND) 7-19, with MK-801 (MK+) or the less active isomer of MK-801 (MK-) (0.25 mg/kg), and trained at either PND 22 or 60. Rats treated with MK+ or MK- and trained on PND 22 were significantly impaired in PSA when compared with the saline control. Beyond the learning impairment, MK+ rats showed an overall decreased running speed during training. They also presented an array of abnormal behaviors and significant weight loss. These nonassociative variables were determined for several doses (0.025, 0. 05, 0.1, 0.15, and 0.20 mg/kg) through PND days 22-25. Rats that received the threshold dose for secondary effects (0.025 mg/kg) also showed an overall decrease in running speed, but failed to show a significant nonspatial learning impairment on PSA. The PSA learning impairment was found to be not long lasting; rats trained at PND 60, after neonatally receiving the original high dose of MK-801, did not differ from controls. PMID- 9736753 TI - Transient expression and transport of brain-derived neurotrophic factor in the male zebra finch's song system during vocal development. AB - The distribution of brain-derived neurotrophic factor (BDNF) in the song system of male zebra finches changes with posthatching age. At day 20, the hyperstriatum ventrale, pars caudale is the only song nucleus in which neurons showed BDNF immunoreactivity. At day 45, the staining in hyperstriatum ventrale, pars caudale was denser than at day 20 and the robust nucleus of the archistriatum, another song nucleus, showed BDNF labeling. By day 65, two additional song nuclei, area X and the lateral magnocellular nucleus of the anterior neostriatum, have become immunoreactive. In the adult, however, the amount of BDNF labeling in all of these brain nuclei is sharply reduced. These sequential events, the anatomical connections between these song nuclei, and the labeling of relevant axons and terminals suggest anterograde transport of BDNF. Furthermore, the timing of BDNF expression coincident with the development of singing behavior suggests that this neurotrophin may be directly involved with the differentiation of the song system. PMID- 9736755 TI - Expression of a dopamine D2 receptor-activated K+ channel on identified striatopallidal and striatonigral neurons. AB - One view of the efferent circuitry of the basal ganglia holds that dopamine D1 and D2 receptors are segregated to striatonigral and striatopallidal neurons, respectively. The present studies investigated whether functional D2-like receptors are, in fact, restricted to striatopallidal neurons. Single, freshly dissociated cells from rat striatum were identified as either striatonigral or striatopallidal projection neurons by fluorescence retrograde labeling. By using cell-attached patch-clamp recordings, neurons of each efferent group were evaluated for the presence of a D2-like receptor-activated 85-pS K+ channel as a measure of receptor function. We now report the presence of this D2 receptor mediated response on both striatal efferent populations, but we observed an approximately 2-fold higher likelihood of encountering the channel on pallidal- versus nigral-projecting neurons. The channel's conductance properties appeared identical in both groups of neurons, but there was a significantly greater open probability for channels detected on striatopallidal neurons. These results indicate that functional D2 receptors are not segregated to striatopallidal neurons, but may be expressed in a higher proportion of, or at a higher density and/or efficiency of coupling on, pallidal- versus nigral-projecting striatal efferents. PMID- 9736756 TI - Aggravated decrease in the activity of nucleus basalis neurons in Alzheimer's disease is apolipoprotein E-type dependent. AB - As reported before, the metabolic activity of nucleus basalis neurons is reduced significantly in Alzheimer patients. Because the apolipoprotein E (ApoE) epsilon4 genotype is a major risk factor for Alzheimer's disease (AD), we determined whether the decrease in metabolic activity in nucleus basalis neurons in AD is ApoE-type dependent. The size of the Golgi apparatus (GA) was determined as a measure of neuronal metabolic activity in 30 controls and 41 AD patients with a known ApoE genotype by using an image analysis system in the nucleus basalis of Meynert. A polyclonal antibody directed against MG-160, a sialoglycoprotein of the GA, was used to visualize this organelle. There was a very strong reduction in the size of the GA in the nucleus basalis of AD patients. Furthermore, a strong and significant extra reduction in the size of the GA was found in the nucleus basalis neurons of AD patients with either one or two ApoE epsilon4 alleles compared with Alzheimer patients without ApoE epsilon4 alleles. Our data show that the decreased activity of nucleus basalis neurons in AD is ApoE epsilon4 dependent and suggest that ApoE epsilon4 participates in the pathogenesis of AD by decreasing neuronal metabolism. PMID- 9736757 TI - Active summation of excitatory postsynaptic potentials in hippocampal CA3 pyramidal neurons. AB - The manner in which the thousands of synaptic inputs received by a pyramidal neuron are summed is critical both to our understanding of the computations that may be performed by single neurons and of the codes used by neurons to transmit information. Recent work on pyramidal cell dendrites has shown that subthreshold synaptic inputs are modulated by voltage-dependent channels, raising the possibility that summation of synaptic responses is influenced by the active properties of dendrites. Here, we use somatic and dendritic whole-cell recordings to show that pyramidal cells in hippocampal area CA3 sum distal and proximal excitatory postsynaptic potentials sublinearly and actively, that the degree of nonlinearity depends on the magnitude and timing of the excitatory postsynaptic potentials, and that blockade of transient potassium channels linearizes summation. Nonlinear summation of synaptic inputs could have important implications for the computations performed by single neurons and also for the role of the mossy fiber and perforant path inputs to hippocampal area CA3. PMID- 9736758 TI - Promiscuous coassembly of serotonin 5-HT3 and nicotinic alpha4 receptor subunits into Ca(2+)-permeable ion channels. AB - Serotonin (5-hydroxytryptamine) type 3 receptors (5-HT3R) and nicotinic acetylcholine receptors are structurally and functionally related proteins, yet distinct members of the family of ligand-gated ion channels. For most members of this family a diversity of heteromeric receptors is known at present. In contrast, known 5-HT3R subunits are all homologs of the same 5-HT3R-A subunit and form homopentameric receptors. Here we show, by heterologous expression followed by immunoprecipitation, that 5-HT3R and nicotinic acetylcholine receptor alpha4 subunits coassemble into a novel type of heteromeric ligand-gated ion channel, which is activated by 5-HT. The Ca2+ permeability of this heteromeric ion channel is enhanced as compared with that of the homomeric 5-HT3R channel. Heteromeric 5 HT3/alpha4 and homomeric 5-HT3Rs have similar pharmacological profiles, but distinct sensitivities to block by the antagonist d-tubocurarine. Coassembly of subunits beyond the boundaries of ligand-gated ion channel families may constitute an important mechanism contributing to the diverse properties and functions of native neurotransmitter receptors. PMID- 9736759 TI - Atypical antipsychotic drugs selectively increase neurotensin efflux in dopamine terminal regions. AB - Typical antipsychotic drugs, such as haloperidol and chlorpromazine, increase synthesis of the neuropeptide neurotensin (NT) in both the striatum and the nucleus accumbens, whereas atypical antipsychotic drugs, such as clozapine and olanzapine, do so only in the nucleus accumbens. By using in vivo microdialysis, we now report that acute administration of haloperidol, clozapine, or olanzapine failed to alter the release of NT in either the striatum or nucleus accumbens. In contrast, chronic administration of haloperidol for 21 days increased NT release in both the striatum and nucleus accumbens, whereas treatment for 21 days with the atypical antipsychotic drugs, clozapine or olanzapine, increased NT release selectively in the nucleus accumbens. These findings suggest that (i) increased NT mRNA expression and NT tissue concentrations are associated with increases in the extracellular fluid concentrations of the peptide and (ii) atypical antipsychotic drugs may exert their therapeutic effects and produce fewer side effects by virtue of their selectivity in limbic compared with striatal, target neurons. PMID- 9736761 TI - One face of a transmembrane helix is crucial in mechanosensitive channel gating. AB - MscL is a mechanosensitive channel in bacteria that responds directly to membrane tension by opening a large conductance pore. To determine functionally important residues within this molecule, we have randomly mutagenized mscL, expressed the genes in living bacteria, and screened for gain-of-function mutants with hampered growth. Expression of these genes caused leakage of cytoplasmic solutes on little or no hypo-osmotic stress. In excised patches, the mutant channels gated at membrane tensions that are less than that required for the gating of the wild type MscL. Hence, the data suggest that the slowed or no-growth phenotype is caused by solute loss because of inappropriate gating of the channel. Most of the mutations mapped to the first transmembrane domain. When this domain is modeled as an alpha-helix, the most severe mutations are substitutions of smaller amino acids (three glycines and one valine) on one facet, suggesting an important role for this structure in MS channel gating. PMID- 9736760 TI - Activation of the genetically defined m1 muscarinic receptor potentiates N-methyl D-aspartate (NMDA) receptor currents in hippocampal pyramidal cells. AB - Evidence suggests that cholinergic input to the hippocampus plays an important role in learning and memory and that degeneration of cholinergic terminals in the hippocampus may contribute to the memory loss associated with Alzheimer's disease. One of the more prominent effects of cholinergic agonists on hippocampal physiology is the potentiation of N-methyl-D-aspartate (NMDA)-receptor currents by muscarinic agonists. Here, we employ traditional pharmacological reagents as well as m1-toxin, an m1 antagonist with unprecedented selectivity, to demonstrate that this potentiation of NMDA-receptor currents in hippocampal CA1 pyramidal cells is mediated by the genetically defined m1 muscarinic receptor. Furthermore, we demonstrate the colocalization of the m1 muscarinic receptor and the NR1a NMDA receptor subunit at the electron microscopic level, indicating a spatial relationship that would allow for physiological interactions between these two receptors. This work demonstrates that the m1-muscarinic receptor gene product modulates excitatory synaptic transmission, and it has important implications in the study of learning and memory as well as the design of drugs to treat neurodegenerative diseases such as Alzheimer's. PMID- 9736762 TI - Hot spots in cold adaptation: localized increases in conformational flexibility in lactate dehydrogenase A4 orthologs of Antarctic notothenioid fishes. AB - To elucidate mechanisms of enzymatic adaptation to extreme cold, we determined kinetic properties, thermal stabilities, and deduced amino acid sequences of lactate dehydrogenase A4 (A4-LDH) from nine Antarctic (-1.86 to 1 degree C) and three South American (4 to 10 degree C) notothenioid teleosts. Higher Michaelis Menten constants (Km) and catalytic rate constants (kcat) distinguish orthologs of Antarctic from those of South American species, but no relationship exists between adaptation temperature and the rate at which activity is lost because of heat denaturation. In all species, active site residues are conserved fully, and differences in kcat and Km are caused by substitutions elsewhere in the molecule. Within geographic groups, identical kinetic properties are generated by different substitutions. By combining our data with A4-LDH sequences for other vertebrates and information on roles played by localized conformational changes in setting kcat, we conclude that notothenioid A4-LDHs have adapted to cold temperatures by increases in flexibility in small areas of the molecule that affect the mobility of adjacent active-site structures. Using these findings, we propose a model that explains linked temperature-adaptive variation in Km and kcat. Changes in sequence that increase flexibility of regions of the enzyme involved in catalytic conformational changes may reduce energy (enthalpy) barriers to these rate governing shifts in conformation and, thereby, increase kcat. However, at a common temperature of measurement, the higher configurational entropy of a cold adapted enzyme may foster conformations that bind ligands poorly, leading to high Km values relative to warm-adapted orthologs. PMID- 9736763 TI - Differential expression of two isopentenyl pyrophosphate isomerases and enhanced carotenoid accumulation in a unicellular chlorophyte. AB - The enzyme isopentenyl pyrophosphate (IPP) isomerase catalyzes the reversible isomerization of IPP to produce dimethylallyl pyrophosphate, the initial substrate leading to the biosynthesis of carotenoids and many other long-chain isoprenoids. Expression of IPP isomerase, and of two enzymes specific to the carotenoid pathway (lycopene beta-cyclase and beta-carotene-C-4-oxygenase), was followed in the green unicellular alga Haematococcus pluvialis after exposure to high illumination. This alga uniquely accumulates carotenoids in the cytoplasm and in late developmental stages turns deep-red in color because of accumulation of ketocarotenoids in the cytosol. The carotenoid/chlorophyll ratio increased 3 fold in wild type and 6-fold in a precocious carotenoid-accumulating mutant (Car 3) within 24 h after increasing the illumination from 20 to 150 micromol photon m 2.s-1. Two cDNAs encoding IPP isomerase in Haematococcus, ipiHp1 and ipiHp2, were identified. Although otherwise highly similar (95% identity overall), the predicted sequence of ipiHp1 contained a 12-aa region not found in that of ipiHp2. This was reflected by a size difference between two polypeptides of 34 and 32.5 kDa, both of which reacted with an antibody to the product of ipiHp1. We suggest that the 32.5-kDa form is involved with the carotenoid accumulation in the cytoplasm, since the 32.5-kDa polypeptide was preferentially up-regulated by high light preceding the carotenoid increase and only this form was detected in red cysts. PMID- 9736764 TI - Task-dependent influences of attention on the activation of human primary visual cortex. AB - There has been a good deal of controversy over whether attention influences area V1-the first cortical area onto which information from the retina is projected. Attention to motion has been found to modulate monkey area MT and the human homolog of MT/MST. Here we show that activation of V1 by attention to motion is task dependent. Our stimulus consisted of a group of translating random dots superimposed over another group of random dots executing expansion motion. Subjects were instructed to pay attention selectively to the translation, expansion, or neither in particular (passive condition). The activity in the human MT/MST homolog measured by functional magnetic resonance imaging (fMRI) was significantly higher in both the translation and the expansion conditions than in the passive condition, while the activity in area V1 was significantly higher only in the translation condition. These results show that attention to motion modulates area V1, and more interestingly that high-level cognitive processing such as attention may directly or indirectly determine the retroactive extent of feedback within the motion pathway in a manner dependent on the type of motion attended. PMID- 9736765 TI - Both here and there: simultaneous expression of autonomous spatial memories in rats. AB - Foraging rats learned to avoid footshock that was present in a part of a circular arena that was either stable or rotating slowly in a lighted room. The rotation dissociated spatial information in the separate reference frames of the room and arena. After learning to avoid the shocked region in either condition, in the absence of shock, memory for this place was expressed by simultaneous avoidance of an area defined in the reference frame of the room as well as of an area defined in the reference frame of the rotating arena. Spatial memories in these distinct reference frames were acquired, retrieved, and extinguished autonomously. PMID- 9736766 TI - Molecular characterization of the mouse gene encoding cellular retinaldehyde binding protein. AB - PURPOSE: To clone and characterize the mouse gene encoding cellular retinaldehyde binding protein (CRALBP). CRALBP appears to modulate enzymatic generation and processing of 11-cis-retinol and regeneration of visual pigment in the vertebrate visual cycle. Mutations in human CRALBP segregate with autosomal recessive retinitis pigmentosa. METHODS: A genomic clone encompassing the 5' end of the CRALBP gene through exon 6 was isolated from a mouse 129/Sv genomic DNA library. Exons 7 and 8 were PCR amplified from mouse eye cDNA and 129/SvJ genomic DNA. The gene structure was determined by automated DNA sequence analysis. RESULTS: The sequence of 6855 nucleotides was determined, including all 8 exons, 3 introns plus 3932 and 629 bases from the 5'- and 3'-flanking regions, respectively. The lengths of introns 3-6 were determined by PCR amplification. Northern analysis identifies a approximately 2.1 kb transcript in mouse eye; Southern analysis supports a single copy gene. CONCLUSIONS: The mouse CRALBP gene is similar to the human gene; the coding sequence is approximately 87% identical, the non-coding sequence approximately 65% identical. In contrast to the human gene, the mouse gene contains a consensus TATA box. One of two photoreceptor consensus elements important for CRALBP expression in human retinal pigment epithelium is also present in the mouse gene. Additional conserved and species-specific consensus sequences are identified. The mouse CRALBP genomic clones and structure provide valuable tools for developing an in vivo model to study protein function and gene regulation. PMID- 9736767 TI - Expression of a copper-containing amine oxidase by human ciliary body. AB - PURPOSE: To examine the molecular structure and ultrastructural distribution of a novel amine oxidase in human ciliary body. METHODS: Human ciliary bodies were solubilized with a nonionic detergent. The solubilized material was subjected to affinity chromatography with 2B4.14.1, a monoclonal antibody which recognizes a family of ciliary body glycoproteins. Proteins eluted from the affinity column were further separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Peptides produced from a 2B4.14. 1-reactive protein with an approximate molecular weight of 100 kDa were analyzed by Edman degradation. The protein thus identified was further examined by Western blotting and immunoelectron microscopy with anti-peptide antisera. RESULTS: Peptide sequences from the 100 kDa ciliary body protein were identical to the predicted protein sequence of an amine oxidase identified recently in a human placental cDNA library. The identity of the ciliary body protein was confirmed by Western blotting with rabbit antiserum generated against the predicted carboxy-terminal peptide of human placenta amine oxidase. Western blotting under nonreducing conditions and following glycosidase digestion indicated that the native enzyme is a disulfide-linked homodimer with multiple N-linked oligosaccharide side chains. By immunoelectron microscopy, the ciliary body amine oxidase was localized to the plasma membranes of inner epithelial cells. CONCLUSIONS: Human placenta amine oxidase is present on the plasma membranes of ciliary body inner epithelial cells. This finding provides a potential explanation for amine oxidase enzyme activity detected in previous studies of anterior segment tissues. Though the functional role of human placenta amine oxidase in the eye is unclear, it may contribute to the production of H2O2 in aqueous humor. PMID- 9736768 TI - The anhidrotic ectodermal dysplasia gene (EDA) undergoes alternative splicing and encodes ectodysplasin-A with deletion mutations in collagenous repeats. AB - Anhidrotic ectodermal dysplasia (EDA) is an X-linked recessive disorder which affects ectodermal structures. A cDNA encoding a 135 amino acid protein with mutations in 5-10% of EDA patients has been reported. We have built up a complete splicing map of the EDA gene and characterized the longest and what most probably represents the full-length EDA transcript, EDA-A. It encodes a 391 amino acid transmembrane protein with a short collagenous domain, (Gly-X-Y)19, and is highly homologous to the protein mutated in Tabby mice (Ta-A). Four new transcripts that code for truncated proteins lacking the collagenous domain were also detected. The splice variants show different expression patterns in eight tissues analyzed, suggesting a regulatory mechanism for gene expression. The EDA-A form of the protein is transported to the cell membrane and induces rounding of the cells, properties also associated with the 135 amino acid isoform. We have determined the genomic organization and the exon-intron boundaries of the EDA gene. SSCP analysis of the nine exons corresponding to EDA-A allowed the identification of mutations in 12 out of 15 EDA patients. Interestingly, three mutations removed either two or four of the Gly-X-Y repeats without interrupting the reading frame, thus suggesting a functional role for the collagenous domain. Our results will allow mutation diagnostics in the majority of patients. PMID- 9736769 TI - Cloning, genomic organization, alternative transcripts and mutational analysis of the gene responsible for autosomal recessive universal congenital alopecia. AB - Complete or partial congenital absence of hair (congenital alopecia) may occur isolated or with associated defects. The majority of families with isolated congenital alopecia has been reported to follow an autosomal recessive mode of inheritance (MIM 203655). We have previously mapped the gene for autosomal recessive congenital alopecia in a large inbred Pakistani family in which affected persons show complete absence of hair development (universal congenital alopecia) to a 15 cM region on chromosome 8p21-22. Here we report the cloning and characterization of the human homologue of the mouse hairless gene and show that it is located in the critical region on chromosome 8p21-22. Determining the exon intron structure allowed detailed mutational analysis of DNA samples of patients with universal congenital alopecia. We detected a homozygous missense mutation in the Pakistani family and a homozygous splice donor mutation in a family from Oman. In addition, we show that the human hairless gene undergoes alternative splicing and that at least two isoforms generated by alternative usage of exon 17 are found in human tissues. Interestingly, the isoform containing exon 17 is the predominantly expressed isoform in all tissues but skin, where exclusive expression of the shorter isoform was observed. We speculate that this tissue specific difference in the proportion of hairless transcripts lacking exon 17 sequences could contribute to the tissue-specific disease phenotype observed in individuals with isolated congenital alopecia. PMID- 9736770 TI - Characterization of a double homeodomain protein (DUX1) encoded by a cDNA homologous to 3.3 kb dispersed repeated elements. AB - Target genes for the helicase-like transcription factor (HLTF), a member of the SNF/SWI family, were immunoprecipitated from HeLa chromatin fragments with an anti-HLTF antibody. A 182 bp fragment ( HEFT1 ) presented 87% sequence identity with 3.3 kb dispersed repeats from the 4q35 D4Z4 locus linked to facioscapulohumeral muscular dystrophy (FSHD). The HEFT1 loci were, however, not genetically linked to FSHD. Transfection and in vitro binding studies identified within HEFT1 a promoter whose basal activity required a GC box activated by Sp1 or Sp3. A 4.4 kb homologous transcript was found mostly in human skeletal muscle and heart. A 1.2 kb cDNA fragment was cloned that encoded a 170 amino acid protein (DUX1) with two paired-type homeodomains. In vitro translated DUX1 specifically interacted in electrophoretic mobility shift assay (EMSA) with a P5 oligonucleotide (5'-GATCTGAGTCTAATTGAGAATTACTGTAC-3'). DUX1 co-expression activated up to 5-fold transient expression in insect cells of a minimal promoter luciferase construct fused to P5. The presence of 20 kDa DUX1 in vivo in rhabdomyosarcoma TE671 cell extracts was shown by western blotting with a rabbit antiserum raised against a DUX1 peptide. This antiserum suppressed a TE671 protein-P5 complex in EMSA with identical migration as the in vitro translated DUX1-P5 complex. Genomic PCR experiments could not identify a gene fragment linking the HEFT1 and DUX1 sequences, which present one mismatch in their overlapping region. However, a similar gene was found in another 3.3 kb element comprising the HEFT1 promoter and a DUX1 -like open reading frame. In addition, homologous gene sequences were identified in 3.3 kb elements of the D4Z4/FSHD locus, considered until now 'junk' DNA. PMID- 9736771 TI - Adenovirus-mediated transfer of the acid alpha-glucosidase gene into fibroblasts, myoblasts and myotubes from patients with glycogen storage disease type II leads to high level expression of enzyme and corrects glycogen accumulation. AB - Glycogen storage disease type II (GSD II) is an autosomal recessive disorder caused by defects in the lysosomal acid alpha-glucosidase (GAA) gene. We investigated the feasibility of using a recombinant adenovirus containing the human GAA gene under the control of the cytomegalovirus promoter (AdCMV-GAA) to correct the enzyme deficiency in different cultured cells from patients with the infantile form of GSD II. In GAA-deficient fibroblasts infected with AdCMV-GAA, transduction and transcription of the human transgene resulted in de novo synthesis of GAA protein. The GAA enzyme activity was corrected from the deficient level to 12 times the activity of normal cells. The transduced cells overexpressed the 110 kDa precursor form of GAA, which was secreted into the culture medium and was taken up by recipient cells. The recombinant GAA protein was correctly processed and was active on both an artificial substrate 4 methylumbelliferyl-alpha-D-glucopyranoside (4MUG) and glycogen. In GAA-deficient muscle cells, a significant increase in cellular enzyme level, approximately 20 fold higher than in normal cells, was also observed after viral treatment. The transduced muscle cells were also able to efficiently secrete the recombinant GAA. Moreover, transfer of the human transgene resulted in normalization of cellular glycogen content with clearance of glycogen from lysosomes, as assessed by electron microscopy, in differentiated myotubes. These results demonstrate phenotypic correction of cultured skeletal muscle from a patient with infantile onset GSD II using a recombinant adenovirus. We conclude that adenovirus-mediated gene transfer might be a suitable model system for further in vivo studies on delivering GAA to GSD II muscle, not only by direct cell targeting but also by a combination of secretion and uptake mechanisms. PMID- 9736772 TI - Characterization of the myotubularin dual specificity phosphatase gene family from yeast to human. AB - X-linked myotubular myopathy (XLMTM) is a severe congenital muscle disorder due to mutations in the MTM1 gene. The corresponding protein, myotubularin, contains the consensus active site of tyrosine phosphatases (PTP) but otherwise shows no homology to other phosphatases. Myotubularin is able to hydrolyze a synthetic analogue of tyrosine phosphate, in a reaction inhibited by orthovanadate, and was recently shown to act on both phosphotyrosine and phosphoserine. This gene is conserved down to yeast and strong homologies were found with human ESTs, thus defining a new dual specificity phosphatase (DSP) family. We report the presence of novel members of the MTM gene family in Schizosaccharomyces pombe, Caenorhabditis elegans, zebrafish, Drosophila, mouse and man. This represents the largest family of DSPs described to date. Eight MTM-related genes were found in the human genome and we determined the chromosomal localization and expression pattern for most of them. A subclass of the myotubularin homologues lacks a functional PTP active site. Missense mutations found in XLMTM patients affect residues conserved in a Drosophila homologue. Comparison of the various genes allowed construction of a phylogenetic tree and reveals conserved residues which may be essential for function. These genes may be good candidates for other genetic diseases. PMID- 9736773 TI - The mouse Y chromosome interval necessary for spermatogonial proliferation is gene dense with syntenic homology to the human AZFa region. AB - The Delta Sxrb deletion interval of the mouse Y chromosome contains Spy, a spermatogenesis factor gene(s) whose expression is essential for the postnatal development of the mitotic germ cells, spermatogonia. The boundaries of Delta Sxrb are defined by the duplicated genes Zfy1 and Zfy2 and four further genes have previously been mapped within the interval: Ube1y and Smcy, linked with Zfy1 on a contig of 250 kb, and Dffry and Uty, which were unanchored. The interval was estimated to be >450 kb. In order to identify any further gene(s) that may underlie Spy, systematic exon trapping was performed on an extended contig, anchored on Zfy1, which covers 750 kb of the Delta Sxrb interval. Exons from two novel genes were isolated and placed together with Dffry and Uty on the contig in the order Dffry-Dby-Uty-Tspy-Eif2gammay-Smcy- Ube1y-Zfy1. All the genes, with the double exception of Tspy, are X-Y homologous and produce putatively functional, spliced transcripts. The tight linkage and order of Dffry, Dby and Uty was shown to be conserved in deletion intervals 5C/5D of the human Y chromosome by the construction of a contig of human PAC and YAC clones; this represents the first example of syntenic homology between Y chromosomes from two distinct mammalian orders. Interval 5C/5D contains the distal boundary of the AZFa interval, which, like Delta Sxrb, is believed to be necessary for spermatogonial development in the prepubertal testis. Our results therefore show that AZFa and Spy may be encoded by homologous genes. PMID- 9736774 TI - Characterization of genes encoding translation initiation factor eIF-2gamma in mouse and human: sex chromosome localization, escape from X-inactivation and evolution. AB - The Delta Sxrb interval of the mouse Y chromosome is critical for spermatogenesis and expression of the male-specific minor transplantation antigen H-Y. Several genes have been mapped to this interval and each has a homologue on the X chromosome. Four, Zfy1 , Zfy2 , Ube1y and Dffry , are expressed specifically in the testis and their X homologues are not transcribed from the inactive X chromosome. A further two, Smcy and Uty , are ubiquitously expressed and their X homologues escape X-inactivation. Here we report the identification of another gene from this region of the mouse Y chromosome. It encodes the highly conserved eukaryotic translation initiation factor eIF-2gamma. In the mouse this gene is ubiquitously expressed, has an X chromosome homologue which maps close to Dmd and escapes X-inactivation. The coding regions of the X and Y genes show 86% nucleotide identity and encode putative products with 98% amino acid identity. In humans, the eIF-2gamma structural gene is located on the X chromosome at Xp21 and this also escapes X-inactivation. However, there is no evidence of a Y copy of this gene in humans. We have identified autosomal retroposons of eIF-2gamma in both humans and mice and an additional retroposon on the X chromosome in some mouse strains. Ark blot analysis of eutherian and metatherian genomic DNA indicates that X-Y homologues are present in all species tested except simian primates and kangaroo and that retroposons are common to a wide range of mammals. These results shed light on the evolution of X-Y homologous genes. PMID- 9736775 TI - Testicular CFTR splice variants in patients with congenital absence of the vas deferens. AB - The involvement of the five thymidine (5T) variant in intron 8 of the cystic fibrosis membrane regulator (CFTR) gene in congenital bilateral absence of the vas deferens (CBAVD) phenotype has been extensively demonstrated. This variant leads to alternative splicing of the CFTR gene which results in a wild-type transcript and one without exon 9. Little is known about expression of the CFTR gene in the testis. We analysed the level of the aberrantly spliced transcripts in testicular biopsies and correlated it with disease expression. Quantitative RT PCR analysis in testicular biopsies from control and CBAVD patients showed a correlation between the length of the IVS8-6(T) n tract and the level of alternatively spliced transcripts. Results from histological analysis also suggest an involvement of the alternative transcript in the spermatogenic status of patients, leading to a decreased number of mature sperm forms in the tubule. PMID- 9736776 TI - Measured haplotype analysis of the angiotensin-I converting enzyme gene. AB - Linkage and segregation analysis have shown that circulating angiotensin-I converting enzyme (ACE) levels are influenced by a major quantitative trait locus that maps within or close to the ACE gene. The D variant of a 287 bp insertion/deletion (I/D) polymorphism in intron 16 of the gene is associated with high ACE levels and may also be related to increased risk of cardiovascular disease. Multiple variants that are in linkage disequilibrium with the I/D polymorphism have been described, but it is unknown if any of these are directly implicated, alone or in combination with as yet undiscovered variants, in the determination of ACE levels. An analysis of 10 polymorphisms spanning 26 kb of the ACE gene revealed a limited number of haplotypes in Caucasian British families due to strong linkage disequilibrium operating over this small chromosomal region. A haplotype tree (cladogram) was constructed with three main branches (clades A-C) which account for 90% of the observed haplotypes. Clade C is most likely derived from clades A and B following an ancestral recombination event. This evolutionary information was then used to direct a series of nested, measured haplotype analyses that excluded upstream sequences, including the ACE promoter, from harbouring the major ACE-linked variant that explains 36% of the total trait variability. Residual familial correlations were highly significant, suggesting the influence of additional unlinked genes. Our results demonstrate that a combined cladistic/measured haplotype analysis of polymorphisms within a gene provides a powerful means to localize variants that directly influence a quantitative trait. PMID- 9736777 TI - ClC-1 chloride channel mutations in myotonia congenita: variable penetrance of mutations shifting the voltage dependence. AB - Mutations in the ClC-1 muscle chloride channel cause either recessive or dominant myotonia congenita. Using a systematic screening procedure, we have now identified four novel missense mutations in dominant (V286A, F307S) and recessive myotonia (V236L, G285E), and have analysed the effect of these and other recently described mutations (A313T, I556N) on channel properties in the Xenopus oocyte expression system. Mutations V286A, F307S and A313T displayed a 'classical' dominant phenotype: their voltage dependence was shifted towards positive potentials and displayed a dominant-negative effect by significantly imparting a voltage shift on mutant-wild-type heteromeric channels as found in heterozygous patients. In contrast, the recessive mutation V236L also shifted the voltage dependence to positive values, but co-expression with wild-type ClC-1 gave almost wild-type currents. I556N, a mutation found in patients with benign dominant myotonia, drastically shifts the voltage dependence, but only a slight shift is seen when co-expressed with wild-type ClC-1. Thus, the voltage dependence of mutant heteromeric channels is not always intermediate between those of the constituent homomeric channel subunits, a conclusion further supported by mixing different ClC-1 mutants. These complex interactions correlate clinically with various inheritance patterns, ranging from autosomal dominant with various degrees of penetrance to autosomal recessive. PMID- 9736778 TI - Characterization of 19 disease-associated missense mutations in the regulatory domain of the cystic fibrosis transmembrane conductance regulator. AB - In order to gain a better insight into the structure and function of the regulatory domain (RD) of the cystic fibrosis transmembrane conductance regulator (CFTR) protein, 19 RD missense mutations that had been identified in patients were functionally characterized. Nine of these (I601F, L610S, A613T, D614G, I618T, L619S, H620P, G628R and L633P) resulted in aberrant processing. No or a very small number of functional CFTR proteins will therefore appear at the cell membrane in cells expressing these mutants. These mutations were clustered in the N-terminal part of the RD, suggesting that this subdomain has a folding pattern that is very sensitive to amino acid changes. Mutations that caused no aberrant processing were further characterized at the electrophysiological level. First, they were studied at the whole cell level in Xenopus laevis oocytes. Mutants that induced a whole cell current that was significantly different from wild-type CFTR were subsequently analysed at the single channel level in COS1 cells transiently expressing the different mutant and wild-type proteins. Three mutant chloride channels, G622D, R792G and E822K CFTR, were characterized by significantly lower intrinsic chloride channel activities compared with wild-type CFTR. Two mutations, H620Q and A800G, resulted in increased intrinsic chloride transport activities. Finally, T665S and E826K CFTR had single channel properties not significantly different from wild-type CFTR. PMID- 9736779 TI - Gene duplications as a recurrent theme in the evolution of the human pseudoautosomal region 1: isolation of the gene ASMTL. AB - We have isolated a novel gene, ASMTL (acetylserotonin methytransferase-like ), in the pseudoautosomal region (PAR1) on the human sex chromosomes. ASMTL represents a unique fusion product of two different full-length genes of different evolutionary origin and function. One part is homologous to the bacterial maf/orfE genes. The other part shows significant homology to the entire open reading frame of the previously described pseudoautosomal gene ASMT, encoding the enzyme catalysing the last step in the synthesis of melatonin. We have also detected the identity of one exon (1A) of ASMT to exon 3 in yet another pseudoautosomal gene, XE7. The data presented suggest that exon duplication and exon shuffling as well as gene fusion may represent common characteristics in the pseudoautosomal region. PMID- 9736780 TI - CAG repeat expansion in autosomal dominant familial spastic paraparesis: novel expansion in a subset of patients. AB - Autosomal dominant familial spastic paraplegia (FSP) is a genetically heterogeneous neurodegenerative disorder displaying anticipation for which three loci have been mapped to the chromosomal positions 14q11.2-q24.3 (SPG3), 2p21-p24 (SPG4) and 15q11.1 (SPG6). The repeat expansion detection (RED) method has been used to demonstrate expanded CAG repeats in some FSP families that map to SPG4. We analyzed 20 FSP families, including four for which there is evidence for linkage to SPG4, and found that in most cases the repeat expansion detected by RED is due to non-pathogenic expansions of the chromosome 18q21.1 SEF2-1 or 17q21.3 ERDA1 locus. Polymorphic expansions at SEF2-1 and ERDA1 appear frequent and may confound RED studies in the search for genes causing disorders demonstrating anticipation. In six FSP families, however, CAG repeat expansion was detected in a subset of affected and at-risk individuals that did not result from expansion of the SEF2-1 and ERDA1 loci. Overall, 11 of 37 (30%) of the FSP patients with a CAG/CTG repeat expansion are unaccounted for by the SEF2-1 and ERDA1 loci, compared with two of 23 (9%) of the unaffected at-risk individuals and none of 19 controls. In the majority of cases these novel expansions were shorter than those previously reported. PMID- 9736781 TI - Lack of PPCA expression only partially coincides with lysosomal storage in galactosialidosis mice: indirect evidence for spatial requirement of the catalytic rather than the protective function of PPCA. AB - Protective protein/cathepsin A (PPCA) is a pleiotropic lysosomal enzyme that complexes with beta-galactosidase and neuraminidase, and possesses serine carboxypeptidase activity. Its deficiency in man results in the neurodegenerative lysosomal storage disorder galactosialidosis (GS). The mouse model of this disease resembles the human early onset phenotype and results in severe nephropathy and ataxia. To understand better the pathophysiology of the disease, we compared the occurrence of lysosomal PPCA mRNA and protein in normal adult mouse tissues with the incidence of lysosomal storage in PPCA(-/-) mice. PPCA expression was markedly variable among different tissues. Most sites that produced both mRNA and protein at high levels in normal mice showed extensive and overt storage in the knockout mice. However, this correlation was not consistent as some cells that normally expressed high levels of PPCA were unaffected in their storage capability in the PPCA(-/-) mice. In addition, some normally low expressing cells accumulated large amounts of undegraded products in the GS mouse. This apparent discrepancy may reflect a requirement for the catalytic rather than the protective function of PPCA and/or the presence of cell-specific substrates in certain cell types. A detailed map showing the cellular distribution of PPCA in nomal mouse tissues as well as the sites of lysosomal storage in deficient mice is critical for accurate assessment of the effects of therapeutic interventions. PMID- 9736782 TI - Mutations in the Treacher Collins syndrome gene lead to mislocalization of the nucleolar protein treacle. AB - Treacher Collins syndrome (TCS) is an autosomal dominant disorder of craniofacial development, the features of which include conductive hearing loss and cleft palate. The TCS gene ( TCOF1 ), which is localized to chromosome 5q32-q33.1, recently has been identified by positional cloning. Analysis of TCOF1 revealed that the majority of TCS mutations result in the creation of a premature termination codon. The function of the predicted protein, treacle, is unknown, although indirect evidence from database analyses suggests that it may function as a shuttling nucleolar phosphoprotein. In the current study, we provide the first direct evidence that treacle is a nucleolar protein. An antibody generated against treacle shows that it localizes to the nucleolus. Fusion proteins tagged to a green fluorescent protein reporter were shown to localize to different compartments of the cell when putative nuclear localization signals were deleted. Parallel experiments using conserved regions of the murine homologue of TCOF1 confirmed these results. Site-directed mutagenesis has been used to recreate mutations observed in individuals with TCS. The resulting truncated proteins are mislocalized within the cell, which further supports the hypothesis that an integral part of treacle's function involves shuttling between the nucleolus and the cytoplasm. TCS is, therefore, the first Mendelian disorder resulting from mutations which lead to aberrant expression of a nucleolar protein. PMID- 9736783 TI - Simultaneous transfer of mitochondrial DNA and single chromosomes in somatic cells: a novel approach for the study of defects in nuclear-mitochondrial communication. AB - The assembly and function of respiratory-competent mitochondria in eukaryotic cells depends on collaboration between the nuclear and mitochondrial genomes, but the molecular mechanisms underlying such cross-talk are poorly understood. Microcell-mediated chromosome transfer has been used to transfer intact chromosomes from one mammalian cell to another, helping to map loci implicated in different diseases and in the senescence process. In the present work, we show that microcells have a significant number of mitochondria which can be transferred to another cell simultaneously with a limited number of chromosomes. By fusing microcells from a colon carcinoma cell line with a mitochondrial DNA (mtDNA)-less osteosarcoma cell line, we were able to isolate transmitochondrial hybrids containing only one of three selectable chromosomes and mtDNA from the donor cell. The proportion of transmitochondrial hybrids containing one chromosomal marker with respect to the total transmitochondrial hybrids and cybrids was approximately 1% and no hybrids were isolated containing more than one nuclear marker. The genetic data correlated well with the composition and structure of the microcell preparations, which showed the presence of cytoplast like structures and microcells containing mitochondria surrounding the micronuclei. Microcell-mediated mtDNA and chromosome transfer can be used to identify nuclear factors implicated in mtDNA maintenance and gene expression, as well as to investigate nuclear factors which modulate clinical phenotypes in mitochondrial disorders. PMID- 9736784 TI - De novo expansion of intermediate alleles in spinocerebellar ataxia 7. AB - Spinocerebellar ataxia 7 (SCA7) is the eighth neurodegenerative disorder caused by a translated CAG repeat expansion. Normal SCA7 alleles carry from four to 35 CAG repeats, whereas pathological alleles carry from 37 to approximately 200. Intermediate alleles (IAs), with 28-35 repeats in the SCA7 gene are exceedingly rare in the general population and are not associated with the SCA7 phenotype, although they have been found among relatives of four SCA7 families. In two of these families, IAs bearing 35 and 28 CAG repeats gave rise, during paternal transmission, to SCA7 expansions of 57 and 47 repeats, respectively, that were confirmed by haplotype reconstructions in one case and by inference in the other. Furthermore, the four haplotypes segregating with IAs were identical to the expanded alleles in each kindred, but differed among the families, indicating multiple origins of the SCA7 mutation in these families with different geographical origins. Our results provide the first evidence of de novo SCA7 expansions from IAs that are not associated with the phenotype but can expand to the pathological range during some paternal transmissions. IAs that segregate in unaffected branches of the pedigrees might, therefore, constitute a reservoir for future de novo mutations that occur in a recurrent but random manner. This would explain the persistence of the disease in spite of the great anticipation (approximately 20 years/generation) characteristic of SCA7. So far, de novo expansions among the disorders caused by polyglutamine repeats have only been demonstrated in Huntington's disease. PMID- 9736785 TI - Recombinant human acid alpha-glucosidase: high level production in mouse milk, biochemical characteristics, correction of enzyme deficiency in GSDII KO mice. AB - Glycogen storage disease type II (GSDII) is caused by lysosomal acid alpha glucosidase deficiency. Patients have a rapidly fatal or slowly progressive impairment of muscle function. Enzyme replacement therapy is under investigation. For large-scale, cost-effective production of recombinant human acid alpha glucosidase in the milk of transgenic animals, we have fused the human acid alpha glucosidase gene to 6.3 kb of the bovine alphaS1-casein gene promoter and have tested the performance of this transgene in mice. The highest production level reached was 2 mg/ml. The major fraction of the purified recombinant enzyme has a molecular mass of 110 kDa and resembles the natural acid alpha-glucosidase precursor from human urine and the recombinant precursor secreted by CHO cells, with respect to pH optimum, Km, Vmax, N-terminal amino acid sequence and glycosylation pattern. The therapeutic potential of the recombinant enzyme produced in milk is demonstrated in vitro and in vivo. The precursor is taken up in a mannose 6-phosphate receptor-dependent manner by cultured fibroblasts, is converted to mature enzyme of 76 kDa and depletes the glycogen deposit in fibroblasts of patients. When injected intravenously, the milk enzyme corrects the acid alpha-glucosidase deficiency in heart and skeletal muscle of GSDII knockout mice. PMID- 9736786 TI - Segregation of a missense mutation in the microtubule-associated protein tau gene with familial frontotemporal dementia and parkinsonism. AB - Frontotemporal dementia and parkinsonism (FTDP) is the second most common cause of neurodegenerative dementia after Alzheimer's disease. Recently, several kindreds with an autosomal dominant form of FTDP have been reported and in some families the pathological locus was mapped to a 2 cM interval on 17q21-22. The MAPT gene, located on 17q21 and coding for the human microtubule-associated protein tau, is a strong candidate gene, since tau-positive neuronal inclusions have been observed in brains from some FTDP patients. Direct sequencing of the MAPT exonic sequences in 21 French FTDP families revealed in six index cases the same missense mutation in exon 10 resulting in a Pro-->Leu change at amino acid 301. Co-segregation of this mutation with the disease was demonstrated by restriction fragment analysis in two families for which several affected relatives were available. The Pro301Leu mutation was not observed in either 50 unrelated French controls or in 11 patients with sporadic frontotemporal dementia. This mutation, which occurs in the second microtubule-binding domain of the MAPT protein, is likely to have a drastic functional consequence. The observation of this mutation in several FTDP families might suggest that disruption of binding of MAPT protein to the microtubule is a key event in the pathogenesis of FTDP. PMID- 9736787 TI - Hemoperfusion with polymyxin B-immobilized fiber attenuates the increased plasma levels of thrombomodulin and von Willebrand factor from patients with septic shock. AB - AIMS: The present study assessed whether plasma levels of thrombomodulin and von Willebrand factor (vWF) are altered in patients with septic shock and whether treatment with polymyxin B-immobilized fiber (PMX-F) affects these levels. METHODS: Twenty-four patients with septic shock and 20 normal healthy controls were included in this study. Plasma levels of thrombomodulin and vWF were measured by enzyme immunoassay (EIA). The treatments with direct hemoperfusion using PMX-F column on patients with septic shock were repeated twice for 2 h each. Healthy controls were not subjected to hemoperfusion. RESULTS: 13 of 24 patients with septic shock survived (survival rate was 54.2%). Levels of blood endotoxin decreased significantly from 41.2 +/- 4.8 pg/ml at baseline to 13.2 +/- 3.6 pg/ml after direct hemoperfusion. Systolic blood pressure increased significantly from 82 +/- 6 mm Hg at baseline to 118 +/- 12 mm Hg after treatment. The patients with septic shock demonstrated significantly increased plasma levels of thrombomodulin (p < 0.001) and vWF (p < 0.001) compared with those in healthy controls. These increased levels of plasma thrombomodulin and vWF in patients with septic shock decreased significantly after treatment with PMX-F (p < 0.01). CONCLUSION: These data suggest that plasma thrombomodulin and vWF may be related to septic shock and that PMX-F is effective in reducing these factors in patients with septic shock. PMID- 9736788 TI - Is post-dialysis urea rebound significant with long slow hemodialysis? AB - BACKGROUND: According to previous studies, postdialysis urea rebound (PDUR) is achieved within 30-90 min, leading to an overestimation of Kt/V of between 15 and 40% in 3- to 5-hour dialysis. The purpose of the study was to assess the impact of PDUR on the urea reduction ratio (URR), Kt/V and normal protein catabolic rate (nPCR) with long 8-hour slow hemodialysis. METHODS: This study was performed in 18 patients (13 males/5 females), 62.5 +/- 11.7 years of age, hemodialyzed for 3 265 months. Initial nephropathies were: 3 diabetes; 2 polycystic kidney disease; 3 interstitial nephritis; 2 nephrosclerosis; 3 chronic glomerulonephritis, and 5 undetermined. Residual renal function was negligible. The dialysis sessions were performed using 1- to 1.8-m2 cellulosic dialyzers during 8 h, 3 times a week. Blood flow was 220 ml/min, dialysate flow 500 ml/min, acetate or bicarbonate buffer was used. Serial measurements of the urea concentration were obtained before dialysis, immediately after dialysis (low flow at t = 0), and at 5, 10, 20, 30, 40, 60, 90 and 120 min, and before the next session. The low-flow method was used to evaluate the access recirculation, second-generation Daugirdas formulas for Kt/V, and Watson formulas for total body water volume estimation. The difference between the expected urea generation (UG) and urea measured after dialysis (global PDUR) defines net PDUR (n-PDUR). RESULTS: The n-PDUR usually became stable after 58 +/- 25 (30-90) min. Its mean value was 17 +/- 10% of the 30-second low-flow postdialysis urea (3.9 +/- 2 mmol/l). This small postdialysis urea value and the importance of UG in comparison with shorter dialysis justify the use of n-PDUR. Ignoring n-PDUR would lead to a significant 4% overestimation (p < 0.001) of the URR (79 +/- 7 vs. 76 +/- 8%), 12% of Kt/V (1.9 +/- 0.4 to 1.7 +/- 0.38) and 4% of the nPCR (1.1 +/- 0.3 to 1.05 +/- 0.3). n-PDUR correlated negatively with postdialysis urea (r = 0.45 p = 0.05), positively with URR (r = 0.31 p = 0.01) and Kt/V (r = 0.3 p = 0.03) but not with K, and negatively with the urea distribution volume (r = 0.33 p = 0.05). Mean total recirculation, ultrafiltration rate, predialysis urea levels and urea clearance did not correlate with n-PDUR. CONCLUSION: We found a significant PDUR in long-slow hemodialysis after a mean of 1 h after dialysis. This PDUR has a less important impact upon dialysis delivery estimation than short 3- to 5-hour hemodialysis, especially for the lower Kt/V or URR ranges. This is explained by the low-flux, high-efficiency, and long-term dialysis. Its inter-individual variability incites us to calculate PDUR on an individual basis. PMID- 9736789 TI - Blood pressure control and hemodynamic changes in patients on long time dialysis treatment. AB - In dialysis patients blood pressure can be well controlled with long dialysis (3 times a week for 8 h) in contrast to a more common short dialysis regime (3 times a week for 4 h). We studied whether the good blood pressure control in patients on long dialysis as compared to patients on short dialysis was associated with a decrease in extracellular fluid volume. Two-day interdialytic ambulatory blood pressure monitoring was performed in 26 non-diabetic patients on long dialysis, in 22 patients on short dialysis, matched for the years they were on dialysis treatment, and during 24 h in 19 healthy volunteers. After full equilibration, 24 h after dialysis, echography of the inferior caval vein was performed to determine fluid state. Cardiac dimensions and stroke index were measured by echocardiography. A blood sample was drawn for the determination of electrolytes and vasoactive hormones. 73% of the patients on short dialysis were using antihypertensive medication in contrast to none of the patients on long dialysis. However, blood pressure was significantly lower in patients on long dialysis (115 +/- 21/67 +/- 11 mm Hg) when compared to patients on short dialysis (143 +/- 26/81 +/- 16 mm Hg). Indexed caval vein diameter, left ventricular diameter index, and atrial natriuretic peptide were not significantly different in patients on long dialysis compared to patients on short dialysis. Also the cardiac index was comparable in patients on long and short dialysis. However, the total peripheral resistance index was significantly lower in patients on long dialysis compared to the patients on short dialysis and normal controls. The left ventricular mass index was increased in both patients on long and short dialysis compared to controls. We conclude that patients on long dialysis have adequate blood pressure control that seems mainly to be caused by a low total peripheral resistance. These data also suggest that factors other than a lower fluid state contribute to the good blood pressure control in patients on long dialysis. PMID- 9736790 TI - New polyether sulfone dialyzers attenuate passage of cytokine-inducing substances from pseudomonas aeruginosa contaminated dialysate. AB - The use of bicarbonate dialysate and high-flux and reprocessed dialyzers has raised concerns about the reverse transfer of dialysate contaminants into the blood compartment. This in vitro study was performed to investigate the reverse transfer of soluble Pseudomonas aeruginosa bacterial products across a polyether sulfone (PES), a newly developed synthetic polymer dialyzer. In vitro dialysis was carried out at 37 degreesC in a closed countercurrent recirculating loop dialysis circuit with a new PES dialyzer. An equal mixture of heparinized whole blood (from healthy volunteers) with pyrogen-free tissue culture medium was circulated in the blood compartment, and bicarbonate dialysate was circulated in the dialysate compartment. After 15 min of dialysis, the dialysate was challenged sequentially with 10(-4), 10(-3), and 10(-2) dilutions of a P. aeruginosa culture supernatant. 1-ml samples were drawn from the blood compartment 5 and 15 min after each challenge and incubated upright at 37 degrees C. At the end of 24 h, Triton X-100 was added, in order to extract total interleukin (IL) 6 and IL-8 production by the whole-blood mixture. These cytokines were measured by electrochemiluminescence assays. At dilutions of 10(-4) and 10(-3), the reverse transfer of soluble bacterial products across the dialyzer was negligible. Five and 15 min after contaminating the dialysate with the highest concentration (10( 2) dilution), the increase in IL-6 production was 239 +/- 170% (p = 0.06) and 886 +/- 444% (p = 0.02), respectively. However, comparing the IL-6-inducing potency of the 10(-2) bacterial supernatant dilution to the spontaneous IL-6 production in the blood compartment during dialysis with the same dilution of dialysate contaminant, there was a dramatic reduction in IL-6 production by 94 and 89% at 5 and 15 min, respectively. Similarly, 5 and 15 min after contaminating the dialysate with the 10(-2) dilution, the increase in IL-8 production was 357 +/- 147% (p = 0.07) and 630 +/- 229% (p = 0.04), respectively. However, comparing the IL-8-inducing potency of the 10(-2) bacterial supernatant dilution to the spontaneous IL-8 production in the blood compartment during dialysis with the same dilution of dialysate contaminant, there was a dramatic reduction in IL-8 production by 93 and 92% at 5 and 15 min, respectively. These results demonstrate that PES dialyzers markedly attenuate passage of cytokine-inducing substances from contaminated dialysate, using a method that detects the entire cytokine synthetic output in the blood compartment. PMID- 9736791 TI - Hemostatic problems in uremia. PMID- 9736792 TI - Disturbed nitric oxide/endothelin-1 equilibrium in cultured human placental endotheliocytes in preeclampsia. AB - Several observations suggest that endothelial cell dysfunction is a central pathophysiologic event of preeclampsia. Endothelin-1 (ET-1) is a vasoconstrictor peptide of the endotheliocyte and is increased in the sera of preeclamptic patients. The aim of this study was to investigate the possible regulatory mechanisms between ET-1 and nitric oxide (NO) production in cultured placental endotheliocytes. We report that endothelial cells of arterial placental blood vessels show a dysfunctional mechanism of negative feedback regulation of ET-1 production by cGMP or insufficient NO release in response to a stimulus in the form of an increasing ET-1 concentration. PMID- 9736793 TI - Intrauterine levels of human decidua-associated protein (hDP) 200 in normal pregnancy and missed abortion. AB - Levels of human decidua-associated protein (hDP) 200 were measured in amniotic fluid samples, obtained from 32 induced and 21 missed abortions at 9-20 weeks of pregnancy. A 3-fold decrease in the hDP 200 level was observed between 9 and 20 weeks of normal pregnancy. Moreover, no significant difference was observed comparing the level of hDP 200 and the pattern of its decrease in missed and induced abortions. Thus, according to our results, hDP 200, being a monoclonal rheumatoid factor, is probably not required for the continuation of normal pregnancy. PMID- 9736794 TI - Use of energy color Doppler in visualizing fetal pulmonary vascularization to predict the absence of severe pulmonary hypoplasia. AB - OBJECTIVE: The aim of our study was to determine if the assessment of pulmonary vascularization by energy color Doppler during ultrasound examination can predict the absence of pulmonary hypoplasia before birth in situations where it is a high risk. METHODS: In a prospective study of 12 pregnancies presenting a risk of pulmonary hypoplasia (5 early and prolonged premature ruptures of the membranes, 1 diaphragmatic hernia, 1 chylothorax, 1 pulmonary sequestration, 1 omphalocele, 1 anamnios and 2 Potter's syndromes) energy color Doppler was used to visualize pulmonary vascularization. RESULTS: In 10 cases where pulmonary vascularization could be visualized, none of the infants had pulmonary hypoplasia. In the 2 cases of Potter's syndrome where pulmonary vascularization was not visualized there was a pulmonary hypoplasia. CONCLUSION: The visualization of fetal pulmonary vascularization with energy color Doppler in situations with a high risk of pulmonary hypoplasia can predict the absence of severe pulmonary hypoplasia. PMID- 9736795 TI - Plasma and membrane Ca2+ and Mg2+ concentrations in normal pregnancy and in preeclampsia. AB - OBJECTIVE: Changes in intracellular Ca2+ and Mg2+ concentrations seem to be involved in the pathogenesis of preeclampsia, whereas the role of cell membranes has not been studied in detail yet. To investigate the changes in Ca2+ and Mg2+ metabolism in normal pregnancy and preeclampsia, plasma and membrane Ca2+ and Mg2+ concentrations were determined in a clinical study as compared to healthy subjects. STUDY DESIGN: 25 healthy female subjects, 22 untreated healthy pregnant and 20 preeclamptic women were investigated. In each patient, plasma and membrane Ca2+ and Mg2+ content were measured. Ca2+ and Mg2+ concentrations were measured by atomic absorption spectroscopy. Erythrocyte membranes were chosen for membranous Ca2+ and Mg2+ determination. RESULTS: Plasma Mg2+ concentrations were significantly lowered in the healthy pregnant group and the preeclamptic group as compared to controls (p < 0.0001). In erythrocyte membranes, Mg2+ content was found significantly decreased in the preeclamptic women as compared to healthy subjects (p < 0.001). In plasma Ca2+ concentrations there was a significant decrease in the preeclamptic group as compared to controls or healthy pregnant women (p < 0.05). Membranous Ca2+ content was significantly increased in the preeclamptic group versus controls or healthy pregnant women (p < 0.001). CONCLUSION: Lowered plasma and membrane Mg2+ concentrations in preeclampsia may contribute to the development of hypertension in pregnancy. Additionally, a disturbed Ca2+ homeostasis is observed in preeclampsia. PMID- 9736796 TI - Prophylaxis of the occurrence of hyperbilirubinemia in relation to maternal oxytocin infusion with steroid treatment. AB - This study was carried out to investigate the steroid prevention on the occurrence and the severity of red blood cell destruction by the effect of oxytocin usage for labor induction. Venous cord blood was collected from the pregnancies who had oxytocin-induced or augmented labors (20), oxytocin-infused deliveries with steroid use (20), deliveries without oxytocin use (20) and cesarean sections (20). Evaluation of the data showed significant increase in serum bilirubin level, serum lactic dehydrogenase activity, erythrocyte fragility and reticulocyte count (p < 0.0083), and a significant decrease in hemoglobulin concentration, packed red cell volume fraction (p < 0.01) in groups with labor induction or augmentation with oxytocin in comparison to deliveries with oxytocin plus steroid use and the two other methods of delivery. Moreover, with regard to the above data, no significant difference was observed between the deliveries other than oxytocin-only use. Mean corpuscular volume in the oxytocin group was apparently (not significant) higher than the steroid group. The results of this study suggest that the use of 16 mg dexamethasone 21-phosphate at the beginning of the induction or augmentation of labor with oxytocin, followed by an additional 4-mg dose 4 h later intravenously, is advantageous for the prevention of erythrocyte destruction. PMID- 9736797 TI - Changes of maternal serum leptin levels during pregnancy. AB - Maternal leptin levels in serum and urine, their relations to maternal weight and body mass index, were examined in 9 healthy pregnant women from the 12th week of gestation until term. Serum leptin concentration was found to increase progressively during the first two trimesters followed by a slight decline thereafter. The peak value of 27.6 +/- 15.3 ng/ml (mean +/- SD) concentration was reached at the 28th week. Serum leptin levels during the first two trimesters correlated significantly with maternal weight (p = 0.002) and body mass index (p = 0.002) but such a relationship was absent during the third trimester. Leptin could be detected only in about half of urine samples; its concentrations proved to be independent of serum values. No correlation was found between maternal serum leptin levels and the birth weight of neonates. Maternal leptin levels appear to refer to alterations in maternal fat tissue mass that occur during pregnancy. PMID- 9736798 TI - Immunolocalisation of interleukin-4 and interleukin-4 receptor in placenta and fetal membranes in association with pre-term labour and pre-eclampsia. AB - The aim of this study was to characterise the localisation and staining intensity of immunoreactive (ir) interleukin-4 (IL-4) and IL-4 receptor (IL-4R) in human placenta and fetal membranes in association with pre-term and term labour, and pre-eclampsia. The data obtained in this study establish the presence of irIL-4 and IL-4R in human placenta and fetal membranes obtained from women at 25-41 completed weeks of gestation. IL-4 and IL-4R immunohistochemical staining was principally localised in villous and chorionic trophoblast, and to amnion epithelial cells. The intensity of IL-4 and IL-4R immunohistochemical staining was not significantly affected by labour status at term or pre-term (p > 0. 05), with the exception of IL-4R in the amnion (p < 0.05). In term amnion, IL-4R was only detectable following labour onset. When data were stratified with respect to the presence or absence of pre-eclampsia, no statistical difference in immunohistochemical localisation or staining intensity for either IL-4 or IL-4R could be identified for any of the tissues studied. Co-localisation of IL-4 and IL-4R within gestational tissues may indicate auto- and/or paracrine mechanisms of action for this cytokine. Tissue-specific, labour-associated induction of IL 4R may contribute to the regulation of biological effects of IL-4 within the uteroplacental unit. PMID- 9736799 TI - Correlation of change in uterine activity to blood loss in the third stage of labour. AB - In 47 women, the change in the uterine activity after the administration of a uterotonic agent was correlated with the amount of blood loss during the same period of time. Uterine activity was measured by a Gaeltec catheter-tipped pressure transducer inserted transcervically within 5 min of delivery of the placenta. A uterotonic agent (either intravenous syntocinon, intramuscular syntometrine or oral misoprostol) was given after the insertion of the intrauterine pressure catheter and pressure recorded for another 90 min. Blood loss over the same 2-hour period was collected with absorbent paper which was then assessed by colorimetric measurement of the haemoglobin content in the sample. Our results show that the change in uterine activity is associated with the total blood loss. However, there is a poor linear correlation between the two variables probably because of the biological variation in myometrial activity and differences in coagulation mechanisms in normal women. PMID- 9736800 TI - Can labor with breech presentation be induced? AB - Our objective was to evaluate the efficacy and safety of labor induction in women with a breech presentation, and an unripe cervix. We conducted a retrospective, matched-paired study on patients with breech presentation and an unripe cervix (n = 23), who underwent induction of labor using extra-amniotic saline instillation. The women were compared to three matched control groups: 46 women with vertex presentation and an unripe cervix, whose labor was induced by the same method, 23 with breech presentation who underwent a vaginal trial of labor, and 23 women with breech presentation who underwent a cesarean section without a trial of labor. In the study group, 12 women (52.2%) delivered vaginally. Rates of Apgar score, birth trauma, and maternal morbidity were similar in all groups. Induction of labor in patients with a breech presentation and an unripe cervix may be attempted in selected cases as it seems to be efficacious (vaginal delivery rate of 52.2%) and safe for both fetus and mother. PMID- 9736801 TI - Insulin sensitivity and sex steroid hormone levels during the menstrual cycle in healthy women with non-insulin-dependent diabetic parents. AB - OBJECTIVE: To identify the effect of the menstrual cycle and sex steroid hormone levels on insulin sensitivity in healthy women with non-insulin-dependent diabetic parents. METHODS: A clinical trial was realized in 6 healthy women with non-insulin-dependent diabetic parents and in 6 control subjects. In both phases of the menstrual cycle the following tests were made: insulin tolerance test, metabolic profile, and sex steroid hormone levels. RESULTS: Insulin sensitivity was significantly lower in the follicular (p = 0.004) and luteal (p = 0.01) phases of the menstrual cycle in probands compared with controls. In the luteal phase dehydroepiandrosterone was higher in probands than in controls (p = 0.009). CONCLUSIONS: Healthy women with non-insulin-dependent diabetic parents had a lower insulin sensitivity in both phases of the menstrual cycle compared with the control group. Dehydroepiandrosterone was higher in the luteal phase in probands than controls. PMID- 9736802 TI - Reproducibility of transvaginal ultrasonographic measurements of endometrial thickness in patients with postmenopausal bleeding. AB - The aim of the study was to establish the reproducibility of transvaginal sonographic measurements of endometrial thickness in patients with postmenopausal bleeding (PMB). In a prospective blind study, two examiners measured the endometrial thickness in 48 patients presenting with PMB by transvaginal sonography on two separate occasions, 30 min apart. The analysis of variance performed at each endometrial thickness measured by the two examiners revealed no statistical difference. However, it was shown that the most accurate measurements are up to the level of 4 mm (mean deviation of 0.1 +/- 0.2 mm, range 0.7). Once the endometrial thickness reaches 5-6 mm the mean deviation becomes 0.3 +/- 1.2 mm with a range of variation of 4 mm. In conclusion, measurements of endometrial thickness in patients presenting with PMB can be repeated quite accurately up to a level of 4 mm thickness. PMID- 9736803 TI - Endosalpingiosis: clinical presentation and follow-up. AB - OBJECTIVE: To define the clinical presentation and follow-up of women found to have endosalpingiosis. METHODS: Subjects included for retrospective study were identified as having had a pathologic diagnosis of endosalpingiosis. Those subjects without coinciding pathologic diagnosis of endometriosis were then identified; their clinical charts were reviewed for information regarding the clinical presentation, surgical findings, therapeutic management, and clinical follow-up. RESULTS: Review of 1,648 pathologic reports from Children's Hospital, Boston and 380 reports from Brigham and Women's Hospital identified 18 subjects with endosalpingiosis without pathologic evidence of endometriosis. Two clinical scenarios were identified, and designated Groups I and II. Group I consisted of 15 patients presenting with pelvic pain, and Group II consisted of 3 patients presenting with infertility, pancreatic cancer, and an abdominal abscess. Follow up was obtained for 8 of the 15 patients in Group I for an average of 17 months, none of whom were pain-free off medications (oral contraceptives, danazol, or GnRH agonist), and 3 (38%) have required additional surgery (6, 10 or 12 months after the initial surgery) at which time all were found to have endometriosis. Follow-up was obtainable for the patient with infertility in Group II for 5 months, with no complaint of pelvic pain. CONCLUSIONS: We report 18 patients with endosalpingiosis without confounding endometriosis. The clinical presentation is one of either pelvic pain or that of an incidental finding; the clinical course and response of those with pain is similar to that of endometriosis. PMID- 9736804 TI - Impact of calcitonin on urinary excretion of prostaglandins during menopause. AB - The aim of this study was to investigate the effects of calcitonin as an antiresorptive agent in postmenopausal osteoporosis in prostaglandin metabolism. Urinary prostaglandin E2 (PGE2) concentrations were determined by PGE2 (125I) RIA kit in a total number of 37 patients in postmenopause; 27 in study group with established osteoporosis (WEO) and 10 in another group without osteoporosis (WO). An additional group of 12 patients in the premenopausal period were selected as controls (PreM). Data were given as mean +/- SD and statistical analysis was performed by Student's t test for paired and unpaired values. A significant decrease in urinary PGE2 concentrations was observed in WEO (7.91 +/- 3.08 vs. 3.79 +/- 3.01 ng/l) (p < 0.001), WO (9.06 +/- 6.76 vs. 6.06 +/- 3.90 ng/l) (p < 0.05) and PreM (7.14 +/- 1.68 vs. 5.16 +/- 1.91 ng/l) (p < 0.01). As a conclusion, calcitonin seems to exert a negative effect on prostaglandin metabolism resulting in reduced new prostaglandin formation. PMID- 9736805 TI - Effects of Ringer's lactate, medroxyprogesterone acetate, gonadotropin-releasing hormone analogue and its diluent on the prevention of postsurgical adhesion formation in rat models. AB - Fifty adult female rats were randomly divided into five groups. Before the standard surgical procedure which consisted of creating a lesion with electrocautery over the uterus, leuprolide acetate (LA) together with its diluent, the diluent alone and medroxyprogesterone acetate (MPA) were injected to the third, fourth, and fifth groups, respectively. Ringer's lactate (RL) solution was applied intraperitoneally to the second group at the end of the surgery. Group I received no medication. Relaparotomy was performed 3 weeks later to evaluate the adhesions. The mean adhesion scores were (mean +/- SD) 2.6 +/- 0.7, 1.4 +/- 0.8, 0.9 +/- 0.6, 2.3 +/- 0.8, and 0.8 +/- 0.6 in groups I, II, III, IV, and V, respectively. The postsurgical adhesions were significantly less in the groups treated with RL, LA, and MPA. PMID- 9736806 TI - A cystocele may compensate for latent stress incontinence by stretching the vaginal hammock. AB - AIMS: To examine why anatomical correction of a cystocele may cause stress incontinence in a patient with no prior history of this condition. METHODS: The study group consisted of 5 patients, aged 45-63 years, with stress incontinence induced by supporting a cystocele with a pessary. The patients were otherwise continent. Testing was performed in a semirecumbent sitting position under perineal ultrasound control. The patients were asked to strain and cough before and after supporting the bladder neck with both sponge-holding forceps and a ring pessary. RESULTS: In all 5 patients, urine loss on stress was noted with bladder base support by forceps and pessary insertion. With both, asymmetrical downward stretching of the vagina and bladder base was observed. When the cystocele was allowed to balloon outwards, the asymmetrical pattern converted to a symmetrical 'funnelling' of the anterior and posterior walls of bladder neck. CONCLUSION: This study appears to support the hypothesis that, in addition to urethral narrowing, a ballooning cystocele may restore urethral closure by removing laxity from the vaginal hammock. Correction of a cystocele may cause incontinence by removing a fortuitous, but abnormal compensatory mechanism. PMID- 9736807 TI - Transforming growth factor-beta1 in hemangioma of the ovary. AB - Hemangioma of the ovary is extremely rare. We report the case of a 32-year-old woman who complained of pelvic pain due to a large right adnexal mass. On surgical exploration a 10 x 8 cm hemangioma of the ovary was resected. Expression of transforming growth factor-beta1 was studied, and the possible role of this molecule in the development of the tumor is discussed. PMID- 9736808 TI - Spontaneous pregnancy after 13 years of amenorrhea in a patient with a voluminous ovarian dysgerminoma and submitted to left adnexectomy and radiotherapy. AB - The authors illustrate the case of a 17-year-old patient who was submitted to left adnexectomy in view of an ovarian dysgerminoma 24 cm in diameter and weighing 2,800 g. She was subsequently submitted to two cycles of radiotherapy. Following a period of amenorrhea lasting 13 years and characterized by high serum levels of gonadotropins, the patient had a spontaneous pregnancy and at 33 weeks of gestation delivered a live and vital fetus. Therefore the occurrence of post radiotherapy amenorrhea, characterized by high serum gonadotropin levels, should not always be considered pathognomonic of precocious menopause. The possibility that radiotherapy causes only a temporary alteration in ovarian activity should also be taken into consideration. PMID- 9736809 TI - Metalloproteinases in the pathogenesis of diabetic nephropathy. PMID- 9736810 TI - Chronic renal failure: an overview from a pediatric perspective. AB - We present data on the costs and impact of chronic renal failure, the primary renal diseases leading to end-stage renal disease in children, and review the adaptive responses and the pathophysiology and complications of uremia in experimental animals and in man. A treatment strategy is summarized. PMID- 9736811 TI - Serum ionized versus total magnesium in patients with chronic renal disease. AB - BACKGROUND: Recent technology has made it possible to assess serum ionized magnesium with user-friendly ion-selective electrodes similar to measurement of the serum calcium concentration. METHODS: We measured the serum ionized (Mg2+) and total magnesium (tMg) concentration in 69 patients with chronic kidney disease (38 men, 31 women, age 22-85 years) and in 75 control subjects. RESULTS: In control subjects the reference ranges were as follows: serum Mg2+ 0. 45-0.67 mmol/l, and tMg 0.67-0.96 mmol/l. In patients with chronic renal failure serum Mg2+ was 0.57 +/- 0.05 mmol/l and tMg 0.80 +/- 0. 11 mmol/l, and in transplant recipients receiving cyclosporine 0.53 +/- 0.05 and 0.71 +/- 0.10 mmol/l, respectively. The correlation coefficient between serum Mg2+ and tMg was r = 0.73 (p < 0.001) in the patient group and r = 0.75 (p<0.001) in control subjects. Serum tMg was below the reference range in 15 of 69 measurements. However, serum Mg2+ was below the reference range in only 1 of these 15 samples. Hence, 14 of 69 cases with low tMg but normal Mg2+ were false-positive with respect to hypomagnesemia. CONCLUSION: We conclude that tMg may overestimate the incidence of hypomagnesemia, and the measurement of Mg2+ may be of benefit when studying patients with expected hypomagnesemia. PMID- 9736812 TI - Plasma soluble fas and soluble fas ligand in chronic glomerulonephritis. AB - It has been reported that glomerular cells with apoptosis and positive Fas immunoreactivity are seen in proliferative glomerulonephritis (PGN). Fas induces apoptosis when it binds to Fas ligand (Fas-L) or soluble Fas-L (sFas-L). However, soluble Fas (sFas) blocks apoptosis by inhibiting binding between Fas and Fas-L or sFas-L. That is, Fas, Fas-L, and sFas-L are inducers of apoptosis, but sFas is an inhibitor of apoptosis. We studied the relationship between the plasma levels of sFas and sFas-L in 32 patients with various types of adult chronic glomerulonephritis. Patients with serum creatinine levels >1.5 mg/dl (132.6 micromol/l) were excluded. The plasma levels of sFas-L were within the normal limits in all patients. The plasma levels of sFas in the patients with minimal change (n = 8) and membranous nephropathy (n = 7) were similar to the age- and sex-matched controls. However, the plasma sFas levels were significantly elevated in patients with mesangial PGN (n = 10) and membranoproliferative glomerulonephritis (n = 7)(3. 4 +/- 0.9 and 3.9 +/- 1.5 ng/ml, respectively) as compared with the age- and sex-matched controls (controls: 2.1 +/- 0.4 and 2.2 +/ 0.6 ng/ml, respectively). In PGN, according to increase of histological grade and decrease of creatinine clearance, the number of TUNEL-positive cells in glomeruli is decreased in spite of an increase of the Fas positivity, and plasma sFas is increased. The degree of proliferative change is determined by the balance between proliferation and apoptosis and/or necrosis. Therefore, increased plasma sFas in PGN may inhibit apoptosis in glomeruli and may be one of the progressing factors in PGN. Thus, we conclude that an increase in plasma sFas levels is important to the protection of apoptosis in PGN. PMID- 9736813 TI - Light chain deposition disease detected by antisera to a variable region of the kappa1 light chain subgroup. AB - A 59-year-old male started maintenance hemodialysis, and 80 months later died of congestive heart failure. Just before death he had no urine, and no monoclonal immunoglobulin could be demonstrated in the serum on immunoelectrophoresis. An autopsy showed deposition of periodic acid-Schiff-positive and Congo red-negative material in the interstitium and vessel walls of almost all organs and tissues. In the glomeruli there was a multifocal nodular accumulation of the material. Electron microscopy showed finely granular, not fibrillar deposits. In spite of these impressive lesions on light microscopy, the material was not stained with commercially available antisera to light chains. However, the material was positive for antisera to the variable region of the kappa1 light chain subgroup. This is the first case with light chain deposition disease that showed positive staining only with antisera to the variable region of the light chain. PMID- 9736814 TI - The serum paraoxonase activity in patients with chronic renal failure and hyperlipidemia. AB - Human serum paraoxonase is physically associated with an apolipoprotein (Apo-A1) and clusterin-containing high-density lipoprotein (HDL) and prevents low-density lipoprotein from lipid peroxidation. The aim of our study was to determine whether paraoxonase activity or phenotype is altered in patients with chronic renal failure and in hyperlipidemic subjects without renal insufficiency and to compare the values with those of healthy controls. We investigated the serum paraoxonase activity and polymorphism in 119 hemodialyzed uremic patients, 107 patients with primary hyperlipoproteinemia, and in 110 healthy control subjects. The serum paraoxonase activity was significantly decreased both in hyperlipidemic (p < 0.01) and uremic patients (p < 0.001) as compared with controls. On comparison, the serum paraoxonase activity was significantly lower (p < 0.001) in uremic than in hyperlipoproteinemic patients. The HDL and Apo-A1 levels were as follows: uremic < hyperlipidemic < control. To assess whether the observed reduction in paraoxonase activity was due to HDL and Apo-A1 level decreases, we standardized the enzyme activity for HDL and Apo-A1 concentrations. We found that the standardized paraoxonase activity (paraoxonase/HDL ratio) was also lower in the uremic patients (103.3 +/- 69.5) as compared with hyperlipidemic patients (137.64 +/- 81.0) and controls (194.45 +/- 94.45). The standardized values for Apo-A1 showed a similar tendency: paraoxonase/Apo-A1 ratio in uremic patients 89.64 +/- 47.8, in hyperlipidemic patients 128.12 +/- 69.83, and in controls 161.40 +/- 47.35. The phenotypic distribution of paraoxonase (AA, AB, BB) did not change significantly in the patient groups. These results suggest that HDL concentration and phenotypic distribution of paraoxonase may not be the only determining factors, but that other as yet undetermined factors could be involved in the enzyme activity changes. PMID- 9736815 TI - Discordance between retinopathy and nephropathy in type 2 diabetes. AB - The aim of this study was to investigate the relationship between the grade of retinopathy and the severity of glomerular lesions in patients with type 2 diabetes and to describe 5 patients without diabetic retinopathy for whom renal biopsy specimens demonstrated advanced diabetic nephropathy. A total of 221 patients with type 2 diabetes (139 males and 82 females) who consectively underwent renal biopsy between 1982 and 1996 were investigated. The severity of diffuse glomerular lesions was graded using the criteria of Gellman and coworkers, and diabetic retinopathy was classified as absent, nonproliferative, or proliferative. The incidence of advanced nephropathy without retinopathy for all 221 cases was 2.3%. Advanced nephropathy was present in 5 of the 122 (4.1%) patients without retinopathy. These 5 patients were all males and aged 50-70 (mean 61) years. Their clinical characteristics were not uniform, and no special clinical features distinguished the patients who were regarded as having possible advanced nephropathy without retinopathy. In our study, although concordance of retinopathy and nephropathy is relatively common, a little discordance was pronounced in Japanese type 2 diabetic patients. Our findings are consistent with the hypothesis that there are important differences in some aspects of the pathogenesis of retinopathy and nephropathy. PMID- 9736816 TI - Effect of an alpha-adrenergic blocker, and ACE inhibitor and hydrochlorothiazide on blood pressure and on renal function in type 2 diabetic patients with hypertension and albuminuria. A randomized cross-over study. AB - alpha-Adrenergic blockers are potential alternative antihypertensive agents for diabetic patients. Data on their relative efficacy and their effect on kidney function and albuminuria are very limited however. 76 patients with diabetes type 2, hypertension (>/=140/90 mm Hg) and albuminuria (>/=30 mg/24 h) were randomized into three groups to receive cilazapril (2.5-10 mg), doxazosin (2-8 mg) or both. Patients of the first and second groups received a single agent for 4 months, the agents were then crossed for an additional period of 4 months followed by the addition of hydrochlorothiazide (25 mg) for a third 4-month period. Blood pressure was monitored monthly, creatinine clearance and HbA1c were measured before and at the end of each treatment period. Patients of the third group received reduced doses of cilazapril and doxazosin for 4 months. Hydrochlorothiazide was then added for the subsequent 4 months. There was a significant decline in blood pressure values during the first period in all groups. Cilazapril: systolic blood pressure (SBP) 160 +/- 6 to 149 +/- 5 mm Hg; diastolic blood pressure (DBP): 101 +/- 3 to 94 +/- 3 mm Hg (p = 0.001). Albuminuria declined from 350 +/- 105 to 205 +/- 96 mg/24 h (p = 0.001), creatinine clearance (CrCl) was unchanged. Doxazosin: SBP: 160 +/- 7 to 151 +/- 6 mm Hg; DBP: 97 +/- 4 to 90 +/- 4 mm Hg (p = 0.001). Albuminuria 373 +/- 121 to 322 +/- 107 mg/24 h (p = 0.065) and CrCl 87 +/- 7 to 91 +/- 6 ml/min. The combination of both agents at half doses was equipotent or superior to either drug alone. Cross-over of cilazapril and doxazosin reproduced the hypotensive effect and reversed the antialbuminuric effect. The addition of hydrochlorothiazide resulted in a further decline of 6-14 mm Hg in SBP and 3-11 mm Hg in DPB. PMID- 9736817 TI - Apo(a) phenotypes and lp(a) concentrations in renal transplant patients. AB - Plasma lipoprotein (a) (LP(a)) concentrations are increased in patients with end stage renal disease. Considering the influence of the apolipoprotein (a) (Apo(a)) polymorphism and the mode of dialysis in this prospective longitudinal study, we compared Lp(a) concentrations before and after the first 6 months of a successful kidney transplantation in 125 recipient patients. Apo(a) phenotyping was performed by using SDS-PAGE and SDS-agarose, isoforms were classified into high molecular weight (HMW) and low molecular weight (LMW). Before the graft, the Lp(a) concentrations were significantly higher in CAPD than in hemodialysis patients (p = 0.021). Six months after transplantation, Lp(a) fell in both treatment groups. This decrease occurred within both LMW and HMW but to a different extent: median relative variations were -35 and -50%, respectively (p = 0. 048). Among patients with Lp(a) concentration >30 mg/dl 6 months after transplantation, 74% had LMW Apo(a) isoform while the remaining 26% had HMW isoform. Successful renal transplantation leads rapidly to a correction of Lp(a) concentrations, especially in patients treated with CAPD who have higher Lp(a) levels. The most important factor seems to be the LMW status corresponding to high Lp(a) levels. PMID- 9736818 TI - Accuracy of noninvasive ultrasonic volume measurements on human kidney transplants. Presentation of a new formula. AB - PURPOSE: To evaluate the precision in volume measurement on human kidney transplants with the aid of noninvasive ultrasonic technique. MATERIAL AND METHODS: A Philips P700 ultrasound machine was used for noninvasive volume determination of 28 porcine kidneys compared to plethysmography. Also, the volume of 46 human transplanted kidneys in situ were measured by ultrasound. Since the configuration of the two types of kidneys differed, a new formula for volume measurement, suitable for kidney transplants, was developed with the aid of clay model experiments. RESULTS: With the standard ellipsoid formula the accuracy of ultrasonic volume determination of the porcine kidneys was good compared with plethysmography, with intra- and interobserver variability of 10.9 and 9.2%, respectively. However, since the width and depth in relation to length of the transplanted kidneys were greater than in the porcine ones (55 vs. 47.5% and 54 vs. 28%, p < 0.01), a new ellipsoid formula was created (r = 1.0, p < 0.0001). CONCLUSION: Noninvasive ultrasonic kidney volume measurements show an acceptable reproducibility (CV 10%). Since the previously used formula for calculating kidney volume was inaccurate on human kidney transplants, a new and accurate ellipsoid formula fitting for human transplanted kidneys is suggested. PMID- 9736819 TI - Evidence of hepatitis C virus passage across dialysis membrane. AB - The passage of hepatitis C virus (HCV) across the dialysis membrane is a controversial issue. We performed a study applying extreme conditions of permeability to the dialysis membrane and avoiding the use of heparin and dialysis bath that might interfere with polymerase chain reaction (PCR) results. We obtained samples from the ultrafiltrate at the beginning of 18 hemodialysis sessions carried out in 6 HCV RNA-positive patients. HCV RNA was detected by PCR in 3 (16.7%) ultrafiltrate samples belonging to 1 of the patients. HCV genotype was the same as that found in positive ultrafiltrate samples and in the serum corresponding to this patient. The viral load of this patient was under the levels detectable by the assay employed. Therefore, contamination of the ultrafiltrate may constitute a potential risk for HCV transmission in hemodialysis units. PMID- 9736821 TI - Abnormal increase of creatine kinase plasma levels following muscle exercise in nephrotic patients. AB - Nephrotic syndrome is a protein-wasting disorder affecting total body protein metabolism, often leading to reduction of lean body mass and changes of muscle cell composition. The aim of this study was to investigate the susceptibility to muscle cell damage in nephrotic patients following submaximal physical exercise, by detection of the creatine kinase (CK) plasma level changes. Fourteen patients affected by primary nephrotic syndrome, without chronic renal failure, underwent an exercise test on a cycle ergometer for 20 min at a constant speed (60 rpm). In each subject, the work rate (expressed as watts) was established as 70% of the maximum power theoretically calculated on a sex, age, weight and height basis. CK plasma levels (U/l) were determined before and 1, 3, 6 and 24 h after the exercise. Following exercise, CK plasma levels became higher in nephrotics than in normal controls. That is, the amount of CK increments was greater in nephrotics than in controls from the first hour after the end of exertion. These changes, both as absolute values and as percentage of the basal values, correlate positively to daily urinary protein losses; moreover, an inverse relationship was detected with albumin serum levels. However, no correlation was observed between the amount of plasma CK increases and age, body weight, plasma creatinine, plasma cholesterol or hemoglobin levels. These results demonstrate that a greater than normal increase of CK plasma levels occurs in nephrotics following physical exercise, and that this increment correlates with the severity of urinary protein wasting. This suggests an increased susceptibility to muscle injury in nephrotic patients probably related to protein depletion and/or to modifications of muscle cell metabolism. Further studies are needed to define the pathogenesis of our findings. PMID- 9736820 TI - Platelet-leukocyte activation during hemodialysis detected with a monoclonal antibody to leukocyte integrin CD11b. AB - Platelet activation is commonly monitored with a battery of monoclonal antibodies against different platelet epitopes with controversial results. The transient expression of platelet markers and their role mediating interactions with other cells could easily explain these discrepancies. The present study has evaluated whether the analysis of a leukocyte activation antigen (CD11b) could provide more reliable results than detection of platelet activation markers. Cytometric techniques with specific monoclonal antibodies were used to compare the reliability of platelet and leukocyte markers to detect activation. Modifications in the presence of platelet glycoproteins GPIb (CD42b), GPIIIa (CD41) and GPIV (CD36), expression of specific platelet markers (P-selectin (CD62P) and lysosomal protein (CD63)) and leukocyte integrin (CD11b) were assessed during hemodialysis. Platelet antigens remained in uremic patients at levels similar or slightly above those detected in a group of healthy subjects. Modifications of platelet antigens during hemodialysis produced inconclusive results. However, numbers of leukocytes expressing CD11b increased progressively during hemodialysis (17.2 +/- 5.1% at 15 min and 21.3 +/- 6.6% at 2 h, p < 0.05, vs. baseline 6.9 +/- 0.2%). The hemodialysis procedure caused an increased formation of leukocyte-platelet aggregates. Detection of leukocyte CD11b may be a useful marker of overall cellular activation during the hemodialysis procedure. PMID- 9736822 TI - Dissociated expression of collagen type IV subchains in diabetic kidneys of KKAy mice. AB - Diabetic nephropathy is characterized by thickening of the glomerular basement membrane and expansion of the mesangial matrix. The glomerular basement membrane is assembled from at least five genetically distinct collagen IV chains. In patients with diabetic nephropathy, differential distribution of these components has been demonstrated. In order to clarify the relationship between progression of diabetic nephropathy and altered type IV collagen assembly in the renal cortex, we examined steady state mRNA levels encoding collagen IV subchains in the kidney cortices of spontaneously diabetic KKAy mice and nondiabetic C57black mice as controls. They were sacrificed at 4, 8, 16, and 24 weeks of age. Northern and dot blot analyses were performed using 32P-labeled mouse probes for classical alpha1(IV) and alpha2(IV) and for alpha3(IV), alpha4(IV), and alpha5(IV) minor chains. The mRNA levels for all collagen IV chains peaked at 4 weeks of age and declined rapidly thereafter in the nondiabetic mice. At all times, alpha1(IV) and alpha2(IV) mRNA expressions were abundant and almost unchanged in KKAy mice. In contrast, mRNA levels for alpha3(IV), alpha4(IV), and alpha5(IV) progressively changed with age. It appears that the expression of minor collagen IV chains is dissociated from the alpha1(IV) and alpha2(IV) chains in diabetic nephropathy. Moreover, an unbalanced increase in the production may affect collagen IV assembly and contribute to basement membrane thickening in diabetic nephropathy of KKAy mice. PMID- 9736823 TI - Enalapril increases antioxidant enzyme activity in renal cortical tissue of five sixths-nephrectomized rats. AB - In rats with five-sixths nephrectomy (remnant kidney), blood pressure, glomerulosclerosis, and proteinuria are significantly reduced by administration of the angiotensin-converting enzyme inhibitor enalapril, during 16 weeks after reduction of the nephron number. The activity of catalase in remnant-kidney cortex homogenate is not influenced by enalapril treatment; the activities of superoxide dismutase and glutathione peroxidase are significantly increased. Elevated lipid peroxidation in cortex homogenates, evaluated by malondialdehyde and 4-hydroxynonenal concentrations, is not changed by treatment. Supplementation of dietary vitamin E to enalapril treatment does not alter antioxidant enzyme activities when compared to enalapril monotherapy. These results show that enalapril improves the balance between reactive oxygen intermediates and antioxidant enzymes in the remnant-kidney cortex of the rat. This finding may in part explain the protective effect of angiotensin-converting enzyme inhibitors on the progression of glomerulosclerosis. PMID- 9736824 TI - Different actions of the cyclooxygenase 2 selective inhibitor flosulide in rats with passive Heymann nephritis. AB - The prostaglandin cyclooxygenase (Cox) exists in two isoforms with different genetic representation. The isoform, which is constitutively expressed (Cox 1), and mediates physiological functions of prostaglandins, and the inducible isoform (Cox 2) which is upregulated by inflammatory stimuli. This study attempts to determine whether a Cox 2 selective inhibitor, flosulide, differs from the mixed type Cox 1 and Cox 2 inhibitor aspirin in respect of renal function and eicosanoid excretion in experimental nephritis. The effects of flosulide and aspirin were studied during the autologous phase of passive Heymann nephritis (PHN) in rats. Female Wistar rats were injected i.v. with 1 ml of Fx1A antiserum at day 1. From day 7 to day 14 they received either aspirin (aspirin, 50 mg/day), flosulide, (0.75 mg/day) or vehicle p.o. The kidney function was evaluated and the animals sacrificed. The kidneys were removed and glomeruli isolated. The glomeruli were incubated in physiological buffer solution. Basal prostaglandin generation was determined in the supernatant. Treatment with flosulide significantly reduced proteinuria as compared to aspirin treatment (64+/-15 vs. 109+/-14 mg/24 h, p < 0.05). Plasma protein and albumin levels were significantly lower in the aspirin-treated group than in flosulide-treated animals (4.7+/-0.26 vs. 5.48+/-0.08 mg/dl, p < 0. 05 and 0.96+/-0.04 vs. 1.25+/-0.10 mg/dl, p < 0.05). Glomerular prostaglandin production (6-keto-PGF1alpha, TxB2, Bicyclo-PGE2) was significantly reduced in aspirin-, but not in flosulide-treated animals. This was mainly due to a reduction of glomerular TxB2 production by aspirin. Our data demonstrate that a Cox 2 selective inhibitor of prostaglandin formation, flosulide, has beneficial effects on preservation of kidney function in rats with PHN, whereas aspirin has not. These beneficial effects of flosulide possibly result from preservation of the physiological glomerular prostaglandin production. Thus, selective Cox 2 inhibitors might be interesting substances for treatment of nephrotic syndrome. PMID- 9736825 TI - Plasma and urinary levels of adrenomedullin in chronic glomerulonephritis patients with proteinuria. AB - In this study, we measured levels of plasma and urinary adrenomedullin (AM) in 37 patients with chronic glomerulonephritis including minimal change nephrotic syndrome, focal segmental glomerulosclerosis or membranous nephropathy that can induce severe proteinuria. Thirty-nine healthy volunteers were enrolled as controls. Plasma and urinary AM levels were measured by an AM-specific radioimmunoassay. Plasma AM concentrations were higher and urinary AM levels were lower in patients with chronic glomerulonephritis than in healthy volunteers. Patients were divided into two groups according to urinary excretion of protein for 24 h (UPro, g/day) which reflects the disease activity or glomerular damage of the glomerulonephritis (group I: Upro < 1, group II: Upro >= 1). Plasma AM levels positively and urinary AM-levels negatively correlated with the degree of proteinuria. These results suggest that plasma and urinary AM levels in patients with chronic glomerulonephritis reflect the disease activity or glomerular damage represented by the degree of proteinuria. PMID- 9736826 TI - Recurrent symptomatic hyponatremia associated with priapism in paraplegia. PMID- 9736827 TI - Plasma adrenomedullin levels, body fluid status, and end-stage renal failure. PMID- 9736828 TI - Minimal-change nephrotic syndrome with acute renal failure associated with missed abortion. PMID- 9736829 TI - Severe hyponatremia probably resulting from inappropriate secretion of antidiuretic hormone. A rare initial presentation of tuberculosis. PMID- 9736830 TI - Effect of angiotensin-converting enzyme inhibitors on hematological parameters and recombinant human erythropoietin doses in peritoneal dialysis patients. PMID- 9736831 TI - Evaluation of the renal tolerance of Xenetix in patients with chronic renal failure. PMID- 9736832 TI - Serum transferrin receptor level in the diagnosis of iron deficiency due to erythropoietin treatment. PMID- 9736833 TI - Hypokalemia, hyperreninemia and osteoporosis in a patient ingesting large amounts of cider vinegar. PMID- 9736834 TI - Keloids and end-stage renal disease. PMID- 9736835 TI - Hepatitis G virus infection among patients undergoing hemodialysis. PMID- 9736836 TI - Audit of professional travel at an academic medical center. PMID- 9736837 TI - A collaborative internship program for premedical students. PMID- 9736838 TI - Requiring students to purchase computers. PMID- 9736839 TI - Another kind of faculty development. PMID- 9736840 TI - Promises to keep. PMID- 9736841 TI - Academic oncology and the forces of the marketplace: crisis cloaks opportunity. PMID- 9736842 TI - Fighting fire with fire: managed-care approaches to behavioral health care in AHCs. PMID- 9736843 TI - Stark choices for AHCs. PMID- 9736844 TI - Health care quality: from data to accountability. AB - The many audiences for information about the quality of health care have different and sometimes conflicting interests and priorities. This is reflected in the diversity of current efforts to use health care data to identify, measure, and demonstrate quality. The author surveys three of these approaches in depth: (1) the professional approach, which relies on the actions of private-sector accreditation groups, trade associations and health plans, hospitals, and other providers to assure quality; (2) the market-driven approach, which relies on the use of quality data by health care purchasers and consumers in choosing plans and providers; and (3) the public-sector approach, which relies on the regulatory, oversight, and purchasing actions of government at the federal, state, and local levels to assure quality. The author concludes that efforts to measure and report the quality of health care invariably confront a variety of technical and political issues. Several observers maintain that it is more important for participants in quality issues to reach consensus on the issues than to reach technical perfection in the way the data are handled. Important obstacles in the technical realm include inadequate investment in sufficiently sophisticated and compatible information systems and the fact that where such systems are in place, they generally cannot be linked. But efforts, both technical and legal, are under way to overcome these obstacles. Even so, some of the issues of health care quality will remain moving targets because of constant changes in the health care environment and in technology. The author closes with the hope that the various actors within the health care industry may coordinate their efforts in dealing with these issues. PMID- 9736845 TI - Ethical considerations for residents. AB - To help residents understand the moral obligations they have undertaken by becoming doctors, the author presents an overview of the ethical landscape of medical practice. She begins by stating that doctors' primary obligation is to use the knowledge of science in working together with others for the good of their patients. This involves (1) relying on the scientific method (and thus eschewing nonscientific alternatives) and supporting or conducting scientific research; (2) embracing the cooperative model (i.e., when appropriate, working cooperatively with other physicians and other health care providers); and (3) working for the good of the patient to preserve life, cure disease, restore or preserve function, educate, and alleviate suffering. In order to fulfill this complex obligation physicians must be professionally competent, they must respect their colleagues and patients, and they must genuinely care about their patients' well-being. The author then discusses the moral complexity of common encounters in medical practice. She explains the ethical conflicts that underlie issues of paternalism, justice, the use of patients for teaching, and end-of-life care. Since new moral problems are introduced with new technology and since medicine is confronting an increasing demand for services in the face of shrinking resources, she maintains that, more than ever, physicians must be aware of the ethical dimensions of their work and be able to organize their understanding of the issues. To meet the complex and demanding commitments of medical practice and to successfully navigate the ethical challenges that they will encounter, physicians must mold themselves not only to be knowledge and skilled professionals but also to be respectful and compassionate human beings. PMID- 9736846 TI - Teaching professionalism: passing the torch. AB - Changes in medicine brought on by health care reform will increasingly pressure physicians and physicians-in-training to adopt business or trade strategies in the name of cost containment and of competition in the health care marketplace. These strategies run directly counter to the professional standards and are a potential threat to medicine's status as a profession. A challenge for this generation of students is not to let this emphasis on finances erode medicine's professionalism. Medical faculty must ensure that their students properly understand the nature of the relationships that permit medicine to enjoy the benefits of being a profession (rather than a trade) and that they learn the appropriate balance between financial and professional considerations. Faculty can and should place financial considerations in proper perspective. Students should learn the basic components of professionalism, how physicians in the past have not always met the full criteria for professionalism, how the current emphasis on cost containment could threaten medicine's status as a profession, appropriate goals for health care reform, the need to form new alliances to meet those goals, and criteria for forming appropriate alliances. Armed with this knowledge, the generation of physicians now in training can understand the delicate balance that must be maintained between financial exigencies and professional imperatives. They will then be prepared to participate in the reform process, embrace its positive aspects, and argue effectively against its negative ones. PMID- 9736847 TI - Cultures in conflict: a challenge to faculty of academic health centers. AB - Academic health centers (AHCs) are experiencing turmoil in all three of their traditional missions of teaching, research, and patient care. The authors examine origins of universities and medical education to place in historical context the stresses affecting AHCs at the end of the 20th century. They describe the cultures of the university to suggest strategies for successful adaptation to these stresses. Clashes of values and norms of the cultures within universities and AHCs can hinder effective adaptation to external change. Administrators, researchers, teachers, and clinicians can have strongly conflicting perspectives. For example, business skill is of increasing importance to the survival of the clinical enterprise, but not typically valued by faculty members. University faculty have often considered accountability as antithetical to academic freedom, and, until recently, accountability was not strongly demanded of AHCs. The authors conclude that AHC faculty must transcend the outdated view that the roles of the scholar, scientist, and healer are in opposition to those of the leader and manager. If AHCs are to survive and prosper through their current cultural transition, their faculty must understand all these roles as part of their intellectual and organizational responsibility. PMID- 9736848 TI - Information about barriers to planned change: a randomized controlled trial involving continuing medical education lectures and commitment to change. AB - PURPOSE: To determine whether practicing physicians receiving only clinical information at a traditional continuing medical education (CME) lecture (control group) and physicians receiving clinical information plus information about barriers to behavioral change (study group) would alter their clinical behaviors at the same rate. METHOD: In a randomized controlled trial, the investigators matched 13 pairs of U.S. and Canadian medical schools, assigning one school from each pair to study or control conditions. Following the commitment-to-change model, the investigators asked the primary care physicians attending control or study lectures on the management of cardiovascular risks whether they intended to make behavioral changes as a result of participating in the lectures and, if so, to indicate the specific changes. Thirty to 45 days later, the investigators surveyed the responding physicians to learn whether they had implemented those changes. RESULTS: Information about barriers to change did not increase the likelihood that physicians in the study group would report successful changes; they were no more likely to change than those in the control group. However, the physicians in both study and control groups were significantly more likely to change (47% vs 7%, p < .001) if they indicated an intent to change immediately following the lecture. CONCLUSIONS: Successful change in practice may depend less on clinical and barriers information than on other factors that influence physicians' performances. To further develop the commitment-to-change strategy in measuring the effects of planned change, it is important to isolate and learn the powers of individual components of the strategy as well as their collective influence on physicians' clinical behaviors. PMID- 9736849 TI - Organizational environment and perceptions of teaching quality in seven South Carolina family medicine residency programs. AB - PURPOSE: To explore the relationship between organizational environment and teaching quality in seven family medicine residency programs. METHOD: In 1995, a questionnaire on organizational environment was administered to the faculties at all seven family medicine residency programs in South Carolina. Eighty-seven percent of the faculty members participated, as did convenience samples of residents, nurses, and administrative staff. The questionnaire measured seven variables: teaching quality, job satisfaction, organizational climate, employees' autonomy, goal attainment, organizational commitment, and job-related stress. RESULTS: Residents, nurses, and administrative staff who were connected to programs at which faculty expressed high levels of job satisfaction assessed teaching quality as higher than did those at other programs. The residents' perceptions of teaching quality were positively correlated with high ratings of organizational climate and job-related stress. The staffs' ratings of goal attainment were also associated with teaching quality. Faculty satisfaction was associated with their reported employee autonomy and goal attainment. CONCLUSION: The organizational characteristics of family medicine residency programs significantly influence the perceptions of teaching quality: specifically, there perceptions are correlated with the degree to which faculty are satisfied with their work environments. In addition, residents' and staffs' perceptions of teaching quality are associated with their attitudes toward their organizations' environments. PMID- 9736851 TI - Effect of medical record audit and feedback on residents' compliance with preventive health care guidelines. AB - PURPOSE: To examine the effect on residents' compliance with preventive health guidelines of an intensive quality-improvement program using medical record audits and individualized feedback. METHOD: The before-and-after study was set in a general internal medicine clinic at a military teaching hospital. In 1995, the authors retrospectively reviewed 280 medical records to determine whether, after the hospital had started an audit-and-feedback program in 1994, residents' compliance rates had risen for preventive health interventions. The study looked at both audited and non-audited interventions. RESULTS: The residents' compliance rates significantly improved for the audited interventions (tetanus immunizations, breast examination, and rectal examination). They also had higher compliance rates for six of the seven non-audited interventions. CONCLUSIONS: An intensive medical record audit with individualized feedback can produce exceptionally high levels of compliance with preventive care practices among internal medicine residents. Furthermore, the improved compliance is generalizable to other health care measures not directly targeted for audit. PMID- 9736850 TI - A failure to reproduce the intermediate effect in clinical case recall. AB - PURPOSE: To investigate the differences between experts, intermediates, and novices in diagnosing and representing clinical cases under various time constraints. METHOD: Second-, fourth-, and sixth-year medical students, and internists studied, diagnosed, and recalled four clinical cases from internal medicine. Participants were allowed to study each case for either 3 minutes, 1 minute 15 seconds, or 30 seconds. The study replicated in most ways the 1993 clinical case recall study of Schmidt and Boshuizen. RESULTS: As expected, diagnostic accuracy increased with level of expertise. However, this study failed to disclose the intermediate effect in clinical case recall that was found in the original study. Instead, a positive linear relation between expertise level and case recall was found. The discrepancy resulted from more elaborate recall by experts in the present study. Constraining processing time did not effect diagnostic accuracy, but equally affected the recall performances of the participants of all expertise levels. This contrasts with the earlier finding that experts' recall is independent of processing time. CONCLUSION: Although it is unclear why the experts' case processing was more elaborate in the present study than in the earlier study, it must be concluded that expert medical knowledge is so flexibly organized that experts can easily represent clinical cases in either the encapsulated or the elaborated mode. PMID- 9736852 TI - Breast cancer screening knowledge and skills of students upon entering and exiting a medical school. AB - PURPOSE: To assess needs for breast cancer screening education by comparing medical students' training and knowledge of breast cancer screening upon their entry to and exit from medical school. METHOD: Seventy-seven medical students at one medical school completed questionnaires as first-year students (in 1992) and again as fourth-year students (in 1996) that assessed their breast cancer screening knowledge. The fourth-year questionnaire included additional questions about the students' clinical training in breast cancer screening skills and their perceptions of needs for further training. RESULTS: Although the students performed significantly better on the knowledge-based questions in their fourth year than they did in their first year, considerable room for improvement remained. The students reported learning the most from surgery rotations and more from standardized patients than from faculty. Women medical students performed significantly more clinical breast examinations than did men students. CONCLUSIONS: Most of the medical students reported needing additional training in clinical breast examination. More curricular time devoted to education about breast cancer screening is needed. PMID- 9736853 TI - The relationship between verbal abuse of medical students and their confidence in their clinical abilities. AB - PURPOSE: To explore the relationship between graduating students' self-reported experiences of verbal abuse during medical school and their confidence in their clinical skills. METHOD: Using data from the Association of American Medical Colleges Medical School Graduation Questionnaire and its Matriculating Student Questionnaire, the author determined the statistical relationships between students' experiences of verbal abuse and their levels of confidence upon graduating medical school. The author controlled for sex, race, age, academic ability (as measured by MCAT scores), and level of assuredness (as measured by levels of confidence upon matriculation). RESULTS: The relationship between verbal abuse and lower levels of confidence was significant for all demographic groups and for students with high and low abilities and high and low levels of assuredness. Although the statistical analysis does not prove causation between verbal abuse and lower confidence, the findings show a correlation between the two. CONCLUSION: Medical schools must understand that verbal abuse correlates with students' confidence, regardless of their sex, race, age, or levels of ability and assuredness. School policies must address the problem of verbal abuse of students to avoid lowering students' self-confidence. PMID- 9736854 TI - Validation of an objective structured clinical examination in psychiatry. AB - PURPOSE: To examine the validity of a psychiatry clerkship's objective structured clinical examination (OSCE). METHOD: In 1996, 33 clinical clerks and 17 psychiatry residents at the University of Toronto participated in an eight station OSCE evaluated by psychiatrist-examiners using binary checklists and global ratings. Prior to the OSCE, communication course instructors were asked to rank the clerks on interviewing ability, and faculty supervisors were asked to identify the OSCE stations on which the clerks were likely to do well or poorly. RESULTS: Mean OSCE scores were significantly higher for the residents than for the clerks on global ratings but not on checklists. The communication instructors accurately predicted the clerks' rankings on the global scores but not their scores on the checklists. The faculty supervisors predicted with moderate accuracy the clerks' success on the OSCE stations as measured by the checklists but not by the global ratings. The residents rated the OSCE scenarios as highly realistic. CONCLUSIONS: The evidence of construct and concurrent validity together with high ratings of realism suggest that a psychiatry OSCE can be a valid assessment of clerks' clinical competence. PMID- 9736855 TI - Physician parents' influence over their children's choices of careers in generalist specialties. PMID- 9736856 TI - Effect of remedial tutorial help on students who fail in-course assessments. PMID- 9736857 TI - Historical highlights of the National Institute of Mental Health from 1946 to the present. AB - This introduction briefly highlights NIMH's history from its inception until today. It is not meant as, nor could it be, a detailed history of NIMH but is offered primarily as a perspective by which to read the articles in this supplement. Obviously, the history of any institution is the personal history of its leadership, and contained in this special issue are the personal reminiscences of six of the eight Directors of NIMH, who review their tenures at the Institute from the perspective of the central contributions and advances made by NIMH during the time they served as Director. As such, it is an interesting and informative personalized history of one of the world's great institutions and one that has played and continues to play a central and vital role in this nation's response to its mentally ill citizens. PMID- 9736858 TI - NIMH before (1946-1970) and during the tenure of Director Bertram S. Brown, M.D. (1970-1978): the early years and the public health mission. PMID- 9736859 TI - NIMH during the tenure of Director Herbert Pardes, M.D. (1978-1984): The President's Commission on Mental Health and the reemergence of NIMH's scientific mission. AB - To summarize these 5-1/2 years, I would offer the following. NIMH--which, like the mental health field in general, has focused principally on services and broad social issues in the 1960s and 1970s--was modified to be a more scientific institute focused on basic biology and behavioral science, major clinical disorders, diagnosis, treatment, and epidemiology. NIMH in its entirety regained a high level of respect in the general NIH community and won increasing support from Congress and the Administration. Increasingly positive perceptions of NIMH may have had a positive effect on the recruitment issue in psychiatry; the numbers of U.S. recruits started to turn back upward. After the early assault by the OMB and the Reagan Administration on the NIMH budget, the year 1982 and 1983 led to a more supportive attitude, and so the threat to the vitality of NIMH and to its overall fiscal support relented. Programs in research training and mental health clinical training and the intramural program were sustained along with the preeminent focus on building extramural support. We recognized that support of the intramural program accounted for an unduly high proportion of overall NIMH research expenditures. In response, we set firm policies designed to build the extramural program while maintaining the strength of the intramural program without expanding it. I might note parenthetically that I had the opportunity to chair an Intramural Research Program Planning Committee convened by NIMH. Without anticipating Dr. Hyman's comments regarding this effort, I will say that we found the intramural research program to be a national resource that, with continued emphasis on scientific quality, should contribute greatly to the nation's mental health and scientific goals in the years ahead. Perhaps in the most global terms the era can be remembered as one in which NIMH shifted toward becoming a predominantly research institute with related education programs. On the one hand, we drew some limits regarding what was considered the purview of NIMH, and we focused much more on illness. On the other hand, we retained much of the richness of NIMH and its focus on the relationships between various disciplines, while catalyzing the extraordinary explosion of the capacity to understand brain and behavior and thereby bring greater promise to the effort to control psychiatric disorders. The excitement of the research and the greater enthusiasm of the government, along with NIMH's encouragement of citizen group activity, contributed to destigmatization and set the groundwork for a much stronger overall advocacy effort on behalf of NIMH, which has continued over the last 10 to 15 years. Simultaneously, attempts were made to secure more data regarding the usefulness of psychiatric treatments and their effectiveness. This too would serve us well in terms of a more favorable attitude toward improving insurance through Medicare and through other areas of mental health care reimbursement. It is an honor to have worked at NIMH. The staff members there are superb, and I want to express my thanks to them. The dedication of outstanding federal leaders is one of the powerful assets of this nation and has been central to the many accomplishments of NIMH. PMID- 9736860 TI - NIMH during the tenure of Director Shervert H. Frazier, M.D. (1984-1986): the mental disorder-based reorganization and research on schizophrenia and severe mental disorders. PMID- 9736861 TI - NIMH during the tenure of Director Lewis L. Judd, M.D. (1987-1990): the decade of the brain and the four national research plans. AB - My tenure at NIMH was an exhilarating, heady time of great satisfaction and achievement for all of us at the Institute. I have great affection and loyalty for NIMH, but my fondest memories are of the individuals who led and staffed the Institute's programs while I was there. One of the most gratifying aspects of my tenure was the opportunity to recruit and appoint people to new responsibilities and to interact with and support them as they grew into and beyond their positions of leadership within NIMH. When I left NIMH, I felt that the Institute's managers and staff were unparalleled in their creativity, competence, commitment, loyalty, and sheer hard work on behalf of the Institute and our field. My thanks and deep gratitude genuinely go out to the entire staff at NIMH during my tenure. However, a special debt of gratitude is owed to a group of colleagues and friends who, at my request, carried very heavy responsibilities and excelled in meeting them: Dr. Alan Leshner (Deputy Director of NIMH, now Director of NIDA); Dr. Stephen Koslow, Dr. Stephen Paul, Dr. Jack "Jay" Burke, Dr. David Segal, Dr. Ira Glick, Dr. Ellen Stover, Dr. Irene Levine, Dr. Daryl Kirsch, Dr. Rex Cowdry, Dr. Sam Keith, Dr. Delores Paron, Leroy Goldman, Richard Pine, William Fitzsimmons, Gordon Seidenberg, Lewis Steinberg, Gemma Weiblinger, George Halter, and my invaluable assistant, Margaret Shanley. PMID- 9736862 TI - NIMH during the tenure of Director Frederick K. Goodwin, M.D. (1992-1994): the return of NIMH to NIH and the fight for parity. PMID- 9736863 TI - NIMH during the tenure of Director Steven E. Hyman, M.D. (1996-present): the now and future of NIMH. PMID- 9736864 TI - Comment: abandoning "race" as a variable in public health research--an idea whose time has come. PMID- 9736865 TI - Comment: Ethnic cleansing in the groves of academe. PMID- 9736866 TI - Topic for our times: if we have the money, why is it so hard? PMID- 9736867 TI - White, European, Western, Caucasian, or what? Inappropriate labeling in research on race, ethnicity, and health. AB - The request for scientifically appropriate terminology in research on race, ethnicity, and health has largely bypassed the term White. This and other words, such as Caucasian, are embedded in clinical and epidemiological discourse, yet they are rarely defined. This commentary analyzes the issue from the perspective of the epidemiology of the health of minority ethnic and racial groups in Europe and the United States. Minority groups are usually compared with populations described as White, Caucasian, European, Europid, Western, Occidental, indigenous, native, and majority. Such populations are heterogeneous, the labels nonspecific, and the comparisons misleading. Terminology that reflects the research purpose-for examples, reference, control, or comparison--is better (unlike White, these terms imply no norm, allowing neither writers nor readers to make stereotyped assumptions about the comparison populations. This paper widens the debate on nomenclature for racial and ethnic groups. Many issues need exploration, including whether there is a shared understanding among the international research community of the terms discussed. PMID- 9736868 TI - Racial discrimination and skin color in the CARDIA study: implications for public health research. Coronary Artery Risk Development in Young Adults. AB - OBJECTIVES: This study assessed whether skin color and ways of handling anger can serve as markers for experiences of racial discrimination and responses to unfair treatment in public health research. METHODS: Survey data on 1844 Black women and Black men (24 to 42 years old), collected in the year 5 (1990-1991) and year 7 (1992-1993) examinations of the Coronary Artery Risk Development in Young Adults (CARDIA) study, were examined. RESULTS: Skin color was not associated with self reported experiences of racial discrimination in 5 of 7 specified situations (getting a job, at work, getting housing, getting medical care, in a public setting). Only moderate associations existed between darker skin color and being working class, having low income or low education, and being male (risk ratios under 2). Comparably moderate associations existed between internalizing anger and typically responding to unfair treatment as a fact of life or keeping such treatment to oneself. CONCLUSIONS: Self-reported experiences of racial discrimination and responses to unfair treatment should be measured directly in public health research; data on skin color and ways of handling anger are not sufficient. PMID- 9736869 TI - The entry of underrepresented minority students into US medical schools: an evaluation of recent trends. AB - OBJECTIVES: Recent challenges to affirmative action suggest the need to reassess the status of the admission of underrepresented minority students to US medical schools. METHODS: The Association of American medical colleges provided US medical school enrollment data and characteristics. Five measures of underrepresented minority enrolled and an overall performance scale were constructed for each school. Multivariate regression identified significant overall performance predictors. Predicted and observed values were compared. RESULTS: Underrepresented minority enrollment increased by 43% after 1986, peaked at 2014 in 1994, did not increase in 1995, and decreased by 5% in 1996. Enrollment was associated with increasing federal research funding and with percentage of underrepresented minorities in the sources population P < .001). The 1996 decline was almost entirely limited to public medical schools. Those in California, Texas, Mississippi, and Louisiana accounted for 18% of 1995 enrollment but 44% of the 1996 decline. CONCLUSIONS: Recent gains in medical school enrollment of underrepresented minorities are being reversed, particularly at public institutions. Implications exist for the health of poor, minority, and underserved communities, which are most likely to be cared for by underrepresented minority physicians. PMID- 9736870 TI - Atopy in children and parental social class. AB - OBJECTIVES: This analysis was conducted to determine whether atopic disorders were related to social class in a pediatric population of a former socialist country. METHODS: A cross-sectional study of 2471 schoolchildren was carried out in 1992 and 1993 in 3 towns in the former East Germany. Parents completed a standardized questionnaire regarding health events and lifestyle factors. In addition, skin-prick tests were performed and total serum immunoglobulin (IgE) was determined. RESULTS: Lifetime prevalence rates for atopic disease and rates of allergic sensitization were highest in children from social class III (in which parents had more than 10 years of formal education) and lowest in social class I (less than 10 years of parental education), while rates in social class II (10 years of parental education) were constant at an intermediate level. CONCLUSIONS: The data confirmed the assumption that in formerly socialist countries social inequalities existed under the socialist system, which were reflected by a social gradient in health outcomes. The findings support the hypothesis that increased access to modern lifestyle could be one reason for the increasing rates of atopic disorders during the last 3 decades. PMID- 9736871 TI - The prevalence of homelessness among adolescents in the United States. AB - OBJECTIVES: Homeless adolescents represent one of the nation's most vulnerable populations. This study reports the 12-month prevalence of homeless episodes among US adolescents. METHODS: Personal, audiotaped interviews were conducted in 1992 and 1993 with a representative household sample of 6496 adolescents aged 12 to 17 as part of the Youth Risk behavior Survey sponsored by the Centers for Disease Control and Prevention. Respondents reported whether they had spent the night in any of a variety of locations other than home during the previous 12 months. RESULTS: Altogether, 7.6% of the youths questioned reported that they had spent at least 1 night in youth or adult shelter (3.3%), public place (2.2%), an abandoned building (1.0%), outside 2.2%), underground (0.4%), or with a stranger (1.1%). Boys were much more likely than girls to report having experienced a homeless episode. CONCLUSIONS: This study suggests that homelessness among adolescents is not simply an urban problem and that prevention programs targeting homeless youths should be implemented nationwide. Additional research is needed to assess the frequency and duration of homeless experiences. Future studies of homelessness in the general population should include questions pertinent to adolescents. PMID- 9736872 TI - Entry into primary care and continuity: the effects of access. AB - OBJECTIVES: This study examined the relationship between access and use of primary care physicians as sources of first contact and continuity with the medical system. METHODS: Data from the 1987 National Medical expenditure Survey were used to examine the effects of access on use of primary care physicians as sources of first contact for new episodes of care (by logistic regression) and as sources of continuity for all ambulatory visits (by multi-variate linear regression). RESULTS: No after-hours care, longer office waits, and longer travel times reduced the chances of a first-contact visit with a primary care physician for acute health problems. Longer appointment waits, no insurance, and no after hours care were associated with lower levels of continuity. Generalists provided more first-contact care than specialists acting as primary care physicians, largely because of their more accessible practices. CONCLUSIONS: These data provide support for the linkage between access and care seeking with primary care physicians. PMID- 9736873 TI - Projections of Alzheimer's disease in the United States and the public health impact of delaying disease onset. AB - OBJECTIVES: The goal of this study was to project the future prevalence and incidence of Alzheimer's disease in the United States and the potential impact of interventions to delay disease onset. METHODS: The numbers of individuals in the United States with Alzheimer's disease and the numbers of newly diagnosed cases that can be expected over the next 50 years were estimated from a model that used age-specific incidence rates summarized from several epidemiological studies, US mortality rates, and US Bureau of the Census projections. RESULTS: in 1997, the prevalence of Alzheimer's disease in the United States was 2.32 million (range: 1.09 to 4.58 million); of these individuals, 68% were female. It is projected that the prevalence will nearly quadruple in the next 50 years, by which time approximately 1 in 45 Americans will be afflicted with the disease. Currently, the annual number of new incident cases in 360,000. If interventions could delay onset of the disease by 2 years, after 50 years there would be nearly 2 million fewer cases than projected; if onset could be delayed by 1 year, there would be nearly 800,000 fewer prevalent cases. CONCLUSIONS: As the US population ages, Alzheimer's disease will become an enormous public health problem. interventions that could delay disease onset even modestly would have a major public health impact. PMID- 9736874 TI - Increasing social variation in birth outcomes in the Czech Republic after 1989. AB - OBJECTIVES: This study investigated social variation in birth outcome in the Czech Republic after the political changes of 1989. METHODS: Routinely collected records on singleton live births in 1989, 1990, and 1991 (n = 380,633) and 1994, 1995, and 1996 (n = 286,907) were individually linked to death records. RESULTS: Mean birthweight fell from 3,323 g to 3,292 g (P < .001) between 1989 and 1991 and then increased to 3,353 g by 1996. The gap in mean birthweight between mothers with a primary education and those with a university education, adjusted for age, parity, and sex of infants, widened from 182 g (95% confidence interval [CI] = 169, 19) in 1989 to 256 g (95% CI = 240, 272) in 1996. Similar trends were found for preterm births. Postneonatal mortality declined most among the better educated and the married. The odds ratio for postneonatal death for infants of mothers with a primary (vs university) education, adjusted for birthweight, increased from 1.99 (95% CI = 1.52, 2.60) in 1989 through 1991 to 2.39 (95% CI = 1.55, 3.70) in 1994 through 1995. CONCLUSIONS: Despite general improvement in the indices of fetal growth and infant survival in the most recent years, social variation in birth outcome in the Czech Republic has increased. PMID- 9736875 TI - The development of sex differences in cardiovascular disease mortality: a historical perspective. AB - OBJECTIVES: Little is known about why males have higher cardiovascular disease (CVD) mortality rates than do females. An important factor that has hampered efforts in this regard is the lack of clarity about whether male excess mortality from CVD has existed throughout history. To answer this question, an investigation was conducted of trends in CVD mortality differences between the sexes from the time when data first became available until the present, including the full range of age groups. METHODS: Mortality statistics for CVD in England and Wales from 1861 through 1992 and in the United States from 1900 through 1991 were used. RESULTS: Three stages in the relationship between male and female CVD mortality were found: (1) An early stage of equal male and female mortality, (2) a stage of the appearance of sex differences in mortality, and (3) a stage with consistently present male excess mortality. CONCLUSION: Male excess mortality from CVD has not always been present in the historical record. Further research is needed to elucidate the causes of this excess mortality. PMID- 9736876 TI - The effect of WIC and Medicaid on infant mortality in the United States. AB - OBJECTIVES: This study examine the impact of participation in the Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) and Medicaid on risk of infant death in the United States. METHODS: The 1988 National Maternal and Infant Health Survey was used to consider the risk of endogenous and exogenous death among infants of women participating in WIC and Medicaid during pregnancy and the infant's first year. RESULTS: Participation in the WIC program during pregnancy and infancy was associated with a reduced risk of endogenous and exogenous infant deaths (odds ratios [ORs] = 0.68 and 0.62, respectively). The risk of endogenous death among infants whose mothers participated in Medicaid during pregnancy was equal to that of the privately insured (OR = 1.04). Uninsured infants faced higher risks of endogenous death (OR = 1.42). CONCLUSIONS: These results show that it is important to consider the net effect of WIC and Medicaid participation and to differentiate both the timing of program receipt and cause of death. Evidence suggests that WIC and Medicaid programs have beneficial effects for poor women and their infants. PMID- 9736877 TI - An assessment of US and Canadian smoking reduction objectives for the year 2000. AB - OBJECTIVES: This study assessed whether US and Canadian smoking reduction objectives for the year 2000 are attainable. The United States seeks to cut smoking in its population to 15%; the Canadian goal is 24%. METHODS: Smoking data were obtained for the United States (1974-1994) and Canada (1970-1995) for the overall populations and several age-sex subpopulations. Analyses estimated trends, future prevalences, and the likelihood of goal attainment. Structural time-series models were used because of their ability to fit a variety of trends. RESULTS: The findings indicate that smoking has been declining steadily since the 1970s, by approximately 0.7 percentage points a year, in both countries. Extrapolating these trends to the year 2000, the US prevalence will be 21% and the Canadian prevalence 24%. CONCLUSIONS: If the current trends continue, the Canadian goal seems attainable, but the US goal does not. The US goal is reachable only for 65-to 80-year-olds, who already have low smoking prevalences. It appears that both countries must increase their commitment to population-based tobacco control. PMID- 9736878 TI - Foster care children and family homelessness. AB - OBJECTIVES: This study examined the association between family homelessness and children's placement in foster care. METHODS: The prevalence of homelessness in a random sample of 195 young foster children was examined. RESULTS: Almost half of the birth parents of the foster children had experienced homelessness. Those children were more likely than other foster children to have siblings in foster care and to be place with nonrelatives. CONCLUSIONS: An extremely high prevalence of family homelessness was found among children in foster care. Policy implications of the association between family homelessness and placement into foster care are discussed. PMID- 9736879 TI - The developmental status and adaptive behavior of homeless and low-income housed infants and toddlers. AB - OBJECTIVES: This study describes the development status of 127 homeless and 91 low-income housed infants and toddlers. METHODS: The Bayley Scales of Infant Development and the Vineland Screener were used to gather data. RESULTS: There were no differences between homeless and low-income housed children. However, younger children in both groups performed better than the older children on most summary scores. CONCLUSIONS: Homeless and low-income housed children did not differ in their cognitive and motor skills. However, older children scored lower than younger children on most measures of development status, suggesting that the cumulative effects of poverty may increase with time. PMID- 9736880 TI - The effect of WIC participation on small-for-gestational-age births: Michigan, 1992. AB - OBJECTIVE: This study examined the relationship between enrollment in the Special Supplemental Nutrition Program for women, Infants, and Children (WIC) and delivery of small-for-gestational-age infants. METHODS: WIC records were linked with birth certificates for 1992 full-term births; 41,234 WIC records and 18,34 non-WIC records were identified. Length of participation was defined by gestational age at enrollment. Logistic regression was used to examine the association between WIC participation and small-for-gestational-age births. RESULTS: Odds ratios for small-for-gestational-age birth decreased with increasing length of enrollment in WIC. CONCLUSIONS: Enrollment in WIC is associated with a lower prevalence of small-for-gestational-age deliveries. PMID- 9736881 TI - Evaluating cluster alarms: a space-time scan statistic and brain cancer in Los Alamos, New Mexico. AB - OBJECTIVES: This article presents a space-time scan statistic, useful for evaluating space-time cluster alarms, and illustrates the method on a recent brain cancer cluster alarms in Los Alamos, NM. METHODS: The space-time scan statistic accounts for the preselection bias and multiple testing inherent in a cluster alarm. Confounders and time trends can be adjusted for. RESULTS: The observed excess of brain cancer in Los Alamos was not statistically significant. CONCLUSIONS: The space-time scan statistic is useful as a screening tool for evaluating which cluster alarms merit further investigation and which clusters are probably chance occurrences. PMID- 9736882 TI - Dispensation of emergency contraceptive pills in Michigan Title X clinics. AB - OBJECTIVES: Emergency contraceptive pill dispensation was estimated in Michigan Title X family planning programs. METHODS: Logistic regression and tobit estimation models were used to predict whether and to what extent emergency contraceptive pills are dispensed. RESULTS: Of the 53 programs studies, 32 dispensed emergency contraceptive pills, averaging fewer than one client per month. Total dispensation was skewed toward a few programs, and the contribution of health departments to this total was small. Emergency contraceptive pill services appeared to be randomly distributed across programs, although most dispenser reported having provided the pills for less than 12 months. CONCLUSIONS: Recent policy advances should lead to more consistent emergency contraceptive pill dispensation in Title X programs. PMID- 9736883 TI - Changes in indications for cesarean delivery: United States, 1985 and 1994. AB - OBJECTIVES: The percentages of cesarean deliveries attributable to specific indications (breech, dystocia, fetal distress, and elective repeat cesarean) were computed for 1985 and 1994. METHODS: Data were derived from the 1985 and 1994 National Hospital Discharge Surveys. RESULTS: Dystocia was the leading indication for cesarean delivery in both years. In comparison with 1985, cesareans performed in 1994 that were attributable to dystocia and breech presentation increased, those attributable to fetal distress did not change significantly, and elective repeat cesareans declined. CONCLUSIONS: Studying indications for cesareans can be useful for hospitals, clinicians, and researchers in determining strategies to lower primary and repeat cesarean rates. PMID- 9736884 TI - Hyaline membrane disease is underreported in a linked birth-infant death certificate database. AB - OBJECTIVE: This study compared the Missouri State Department of Health linked birth-infant death certificate database and medical records with respect to recording hyaline membrane disease in very low-birth-weight infants. METHODS: We reviewed the records for all 976 infants weighing 500 to 1500 g who were born to St. Louis, Mo, residents in 1989, 1991, and 1992. RESULTS: Eighteen percent of the birth certificates and 54% of the medical records documented hyaline membrane disease, resulting in 34% sensitivity and 99% specificity. CONCLUSIONS: The Missouri State Department of Health birth-infant death certificate database underestimates the incidence of hyaline membrane disease, which suggest that national statistics for the disease are also underestimated. PMID- 9736886 TI - Telephone coverage and measurement of health risk indicators: data from the National Health Interview Survey. AB - OBJECTIVES: This study compared health behavior variables for all US households and households with telephones to measure the potential impact of telephone coverage on estimates from telephone surveys. METHODS: Data were derived from the 1991 through 1994 version of the National Health Interview Survey. RESULTS: Ninety-five percent of respondents lived in households with telephones. Differences in health indicators were small (< 1%) in comparisons between all households and those with telephones. Results were similar when only respondents below the poverty level were included. CONCLUSIONS: Telephone noncoverage effects appear to be small, supporting the use of telephone surveys for health risk behavior surveillance with most population groups. PMID- 9736885 TI - Reactions of adult and teenaged smokers to the Massachusetts tobacco tax. AB - OBJECTIVES: This study assessed smokers' reactions to a 25 cents cigarette tax imposed in Massachusetts. METHODS: A statewide telephone survey of 1783 adult smokers and 216 teenaged smokers was conducted. RESULTS: Among adult smokers, 3.5% reported that they had stopped smoking, owing in part to the price increase; 35% had considered quitting and 19% had attempted to cut the cost of smoking by switching to cheaper brands or cutting down. Among teenagers, 21% had considered quitting and 26% had cut costs. Low-income smokers were more responsive to the price increase than more affluent smokers. CONCLUSIONS: A modest and temporary price increase promoted quitting among adult smokers and reduced cigarette consumption among low-income teenagers. PMID- 9736887 TI - Social class, ethnicity, and mental illness: the importance of being more than earnest. AB - This paper revisits a landmark study of the prevalence of mental illness in the state of Massachusetts conducted by Edward Jarvis in the 19th century. Jarvis drew an improper conclusion about the relationship between social class, ethnicity, and insanity, asserting that the Irish foreign-born had a higher prevalence of insanity in each social stratum. A reanalysis of Jarvis' data shows that in both the pauper and independent social classes in Massachusetts, the prevalence of insanity was significantly lower among foreign-born persons than among native-born persons. On the basis of his misperception, Jarvis constructed elaborate etiological theories. These theories made a strong impact on the mental health service policies of his day. The effects of incomplete examination of data on etiological theories and mental health policy in current times are highlighted in this article. PMID- 9736888 TI - Hawaii's statewide syringe exchange program. PMID- 9736889 TI - Columbian field epidemiology training program. PMID- 9736890 TI - The neglected lesson of the Tuskegee study. PMID- 9736891 TI - A geographic information systems application for disease surveillance. PMID- 9736892 TI - Performance following ability-focused physical therapy intervention in individuals with severely limited physical and cognitive abilities. AB - BACKGROUND AND PURPOSE: Do individuals with severely limited physical and cognitive abilities improve their gross motor abilities when given physical therapy intervention, and does improvement transfer to nontreatment settings? SUBJECTS: The subjects were 24 individuals (10 female, 14 male), aged 3 to 30 years (X = 20.1, SD = 8.1), who were nonambulatory and had limited adaptive behavior. METHODS: Change in gross motor ability during 18 weeks of twice-weekly therapy was measured using goal attainment scaling (GAS). Three gross motor goals were developed for each subject based on individual or caregiver needs, with one goal randomly selected as a control. Physical impairments were treated, and behavioral management principles, low-level communication approaches, high repetition practice of goals, and a progressive reduction of both physical assistance and multisensory cues were used. An independent rater scored goal level from randomly ordered videotapes recorded during therapy and in recess and home settings. RESULTS: Mean GAS T scores were higher for treatment goals (X = 45.6, SD = 10.5) compared with control goals (X = 34.6, SD = 11.8). When the expected goal level (50) was met during therapy, mean GAS T scores in recess settings ( X = 35.9, SD = 11.5) and home settings (X = 42.2, SD = 12.2) were lower. At the conclusion of therapy, there were no differences in goal levels between treatment and control goals in both the recess and home settings. CONCLUSION AND DISCUSSION: The subjects demonstrated improvement of gross motor abilities practiced during therapy. Level of ability during therapy, however, did not consistently transfer to the recess of home settings. [Brown DA, Effgen SK, Palisano RJ. Performance following ability-focused physical therapy intervention in individuals with severely limited physical and cognitive abilities. PMID- 9736893 TI - Use of generic versus region-specific functional status measures on patients with cervical spine disorders. AB - BACKGROUND AND PURPOSE: Few data exist to support the use of functional status measures on patients with disorders of the cervical spine. This study was designed to compare the construct validity and sensitivity to change of the Neck Disability Index (NDI) and the Medical Outcomes Study 36-Item Short-Form Health Survey (SF-36). SUBJECTS AND METHODS: Patients (N = 146) with a variety of disorders of the cervical spine completed the NDI and the SF-36 prior to treatment. Following discharge from treatment, 69 of these patients completed a second NDI and SF-36. RESULTS: There was evidence for the construct validity and sensitivity to change of the NDI and the physical and mental component scores of the SF-36. The ability of the NDI and SF-36 to detect change varied, depending on the construct tested. The SF-36 was superior in some instances, and the NDI was superior in other instances. The NDI appears to measure both mental and physical health-related factors. CONCLUSION AND DISCUSSION: There appears to be substantial overlap between the 2 measures. The use of both measures, therefore, is probably not necessary. [Riddle DL, Stratford PW. Use of generic versus region specific functional measures on patients with cervical spine disorders. PMID- 9736894 TI - Evaluation of reflex- and nonreflex-induced muscle resistance to stretch in adults with spinal cord injury using hand-held and isokinetic dynamometry. AB - BACKGROUND AND PURPOSE: In this study, we compared the intertrial reliability of resistive torque measurements obtained with hand-held and isokinetic dynamometers and examined the validity of the hand-held dynamometers for the assessment of spastic hypertonia, defined as reflex- and nonreflex-induced resistance to stretch. SUBJECTS: Nine subjects (mean age = 40.6 years) with a chronic (1-5 years) spinal cord injury participated. METHODS: The plantar flexors were stretched at 5 degrees /s (low velocity [LV]) and 180 degrees /s (high velocity [HV]) with an isokinetic dynamometer while the evaluator attempted to match these velocities with a hand-held dynamometer. Electromyographic activity of the soleus and tibialis anterior muscles as well as ankle displacements were recorded. Resistive torque and velocity, measured at -5 degrees of dorsiflexion, were averaged (n = 4). RESULTS: High intraclass correlation coefficients (ICCs) were found at LV and HV for both the hand-held (ICC = .93 and .84) and isokinetic (ICC = .99 and .93) dynamometers. With the hand-held dynamometer, lower resistive torques were found at LV (0.8 N.m) and HV (1.2 N.m), whereas higher velocities were attained at HV. CONCLUSION AND DISCUSSION: The results indicate that the reproducibility of resistive torques obtained with hand-held dynamometry compares with that obtained with isokinetic dynamometry and allows testing of velocities that can be adjusted to the specific level of resistance to stretch. Electromyography confirmed the validity of hand-held dynamometry for assessing reflex and nonreflex components of SH. [Lamontagne A, Malouin F, Richards CL, Dumas F. Evaluation of reflex- and nonreflex-induced muscle resistance to stretch in adults with spinal cord injury using hand-held and isokinetic dynamometry. PMID- 9736895 TI - Reliability of physical examination items used for classification of patients with low back pain. AB - BACKGROUND AND PURPOSE: The purpose of this study was to examine the interrater reliability of measurements obtained by examiners administering tests proposed to be important for classifying low back pain (LBP) problems. SUBJECTS: Ninety-five subjects with LBP (41 men, 54 women) and 43 subjects without LBP (17 men, 26 women) were examined by 5 therapists trained in the techniques used. METHODS: A manual was developed by the first author that described the clinical examination procedures. The therapists were trained by the first author in the test procedures and definitions. The training included instruction through videotapes, practice and a written examination. Each examination was conducted by a pair of therapists. Within a pair, a therapist was the primary examiner for half of the subjects and an observer was the primary examiner for half of the subjects. Examination findings were recorded independently, without discussion. RESULTS: Percentage of agreement and generalized kappa coefficients were used to analyzed the data. Kappa values were > or = .75 for all 28 items related to the symptoms elicited and > or = .40 for 72% of the 25 items related to alignment and movement. CONCLUSION AND DISCUSSION: The results suggest that experienced therapist who had trained together were able to agree on the results of examinations and obtain an acceptable level of reliability. Future work should focus on testing of reliability when more than one therapist performs the examination and when therapist not trained by the test developer to administer the examination perform the tests. [Van Dillen LR, Sahrmann SA, Norton BJ, et al. Reliability of physical examination items used for classification of patients with low back pain. PMID- 9736896 TI - Validity and reliability of measurements obtained with an "activity monitor" in people with and without a transtibial amputation. AB - BACKGROUND AND PURPOSE: In this study, the validity and reliability of measurements obtained with an "Activity Monitor" (AM) were examined. The instrument is designed to monitor ambulatory activity by use of accelerometer signals, and it detects several activities associated with mobility (standing, sitting, lying, transitions, movement-related activities). SUBJECTS: Four men with a transtibial amputation and 4 men without a transtibial amputation participated. METHODS: The subjects performed normal daily activities, during which accelerations were measured and videotape recording were made (reference method). Validity was assessed by calculating agreement scores between the AM output and the videotape recordings and by comparing the number of transitions and the duration of activities determined by both methods. RESULTS: The overall agreement between the AM output and the videotape recordings was 90%. Other agreement scores, in addition to the determination of the number of transitions and the duration of activities, were generally within a range of error of 0% to 10%. CONCLUSION AND DISCUSSION: The reliability and validity of the AM measurements appeared to be good, which supports its potential use in rehabilitation and physical therapy. [Bussmann HBJ, Reuvekamp PJ, Veltink PH, et al. Validity and reliability of measurements obtained with an "Activity Monitor" in people with and without a transtibial amputation. PMID- 9736897 TI - Description of movement patterns of young adults moving supine from the foot to the head of the bed. AB - BACKGROUND AND PURPOSE: The purpose of this study was to describe the movement patterns (MPs) in young adults moving supine from the foot to the head of a bed. SUBJECTS: Thirty-six young adults, aged 19 through 44 years, participated in this study. METHODS: Subjects were videotaped performing 10 trial of moving in bed. The MPs of 3 body regions were described and categorized. RESULTS: Six MP categories were developed for the axial region, 8 MP categories were developed for the upper extremities, and 6 MP categories were developed for the lower extremities. Kappa values ranged from .81 to .90 for the 3 body regions, Fifty nine different combinations of MPs were observed. The most common combination of MPs occurred in 21.2% of the trials, 3 times more frequently than any other combination. CONCLUSION AND DISCUSSION: A variety of MPs are used by young adults for moving from the foot to the head of the bed. A symmetrical pattern of sitting up and pushing with both hands and both feet was found to be the most common pattern. [Cohen BG, Cardillo ER, Lugg D, et al. Description of movement patterns of young adults moving supine from the foot to the head of the bed. PMID- 9736898 TI - Outcomes research and surgeons. PMID- 9736899 TI - Multiple endocrine neoplasia type 2-associated RET proto-oncogene mutations do not contribute to the pathogenesis of sporadic parathyroid tumors. AB - BACKGROUND: Parathyroid disease occurs sporadically or as part of hereditary multiple endocrine neoplasia (MEN) syndrome. The aim of this study was to evaluate the possible role of the RET proto-oncogene not only in hereditary MEN 2 associated hyperparathyroidism but also in different forms of sporadic hyperparathyroidism. METHODS: We investigated 22 patients with parathyroid disease whose family history and results of laboratory and clinical examinations excluded MEN 2 syndrome. DNA extractions of histologically confirmed tumor tissue of patients with primary hyperparathyroidism (n = 18), renal hyperparathyroidism (n = 2), and parathyroid carcinoma (n = 2) were performed. Using solid phase DNA sequencing, mutation analysis of polymerase chain reaction amplified products focused on exons 10, 11, and 16 of the RET proto-oncogene. Parathyroid tissue from four patients with known MEN 2A served as positive controls. RESULTS: No mutations of the codons 609, 611, 618, 620, 634, and 918 were found in the sporadic parathyroid tumors analyzed. DNA sequencing revealed heterozygous mutations in codon 634 of the RET proto-oncogene in four parathyroid glands from four patients with MEN 2A. CONCLUSIONS: Mutations of the RET proto-oncogene contributing to MEN 2 syndromes are absent in sporadic parathyroid tumors. Our data in conjunction with the literature suggest at least three different modes of tumorigenesis in parathyroid disease. PMID- 9736900 TI - Prolonged hypoxia alters endothelial barrier function. AB - BACKGROUND: It is well recognized that hypoxia/reoxygenation and exposure to inflammatory mediators such as cytokines and neutrophils alter the barrier function of the vascular endothelium. The experiments we conducted tested whether hypoxia alone could produce changes in permeability and whether a prolonged period of hypoxia alters the surface expression of cell adhesion molecules. METHODS: Endothelial cells were cultured from human umbilical vein endothelial cells (HUVECs). Hypoxia was created by isolating the cells in a chamber through which 1% 02, 5% CO2, and 94% N2 were insufflated (30 min at 1/min). Oxygen tension was measured through oxygen-quenching phosphorescence. Hypoxia was maintained for 24 hours. Changes in endothelial permeability were measured by transendothelial electrical resistance (TEER). Endothelial leukocyte adhesion molecule 1 (ELAM-1) and intercellular adhesion molecule 1 (ICAM-1) expression were assessed by flow cytometry (mean +/ standard error of the mean [SEM]. RESULTS: Exposure of endothelial cells to hypoxia resulted in increased permeability between 6 and 24 hours, with the greatest decrease in TEER at 18 hours (63% +/ 3%; P < .05). Prolonged hypoxia produced no change in the surface expression of ELAM-1 or ICAM-1. CONCLUSIONS: Hypoxia alone produced a significant reversible alteration in endothelial permeability. However, this change was observed only under severe hypoxic conditions (eg, below 20 mm Hg); higher oxygen tensions (25 and 35 mm Hg) had no significant effect. Unlike observations made after cytokine exposure, hypoxic breakdown of endothelial barrier function was unassociated with up-regulation of either ELAM-1 or ICAM-1. PMID- 9736901 TI - Decrease in prevalence of Buerger's disease in Japan. AB - BACKGROUND: Buerger's disease is a peripheral arterial occlusive disease that is becoming rare in Western countries but is more common in Asia. Whether it is a specific disease entity remains controversial. This study was undertaken to investigate changes in the prevalence and characteristics of Buerger's disease at a major institution in Japan. METHODS: Patients with Buerger's disease admitted to Nagoya University Hospital between January 1985 and December 1996 were studied retrospectively. Buerger's disease was diagnosed on admission according to Shionoya's clinical criteria. RESULTS: A total of 105 patients with Buerger's disease were evaluated on 126 admissions; 58 were new patients who were admitted for initial treatment, and 47 patients were experiencing a worsening of Buerger's disease and had a history of prior treatment. Forty-six new patients were admitted between 1985 and 1989, but only 12 new patients were admitted between 1990 and 1996 (9 +/ 3/ yr vs 2+/ 2/ yr, p = 0.0003). Between 1985 and 1989, 44 patients were admitted because of disease exacerbation, whereas only 24 such admissions occurred between 1990 and 1996 (9+/ 3/ yr vs 3 +/ yr, p = 0.0137). The number of admissions for atherosclerotic peripheral vascular disease did not change significantly in that period. Of the 105 patients, the majority (96%) were men; mean age at the time of disease onset was 36 +/ 8 years. The chief complaint on admission was gangrene/ulcer in 64%, rest pain in 13%, foot claudication in 6%, calf claudication in 6%, and other in 10%. CONCLUSION: The prevalence of Buerger's disease appears to be decreasing at our institution in Japan. Its clinical characteristics have not changed. A similar decrease in prevalence appears to have occurred in Western countries. PMID- 9736902 TI - Major histocompatibility complex class II expression and parathyroid autoantibodies in primary hyperparathyroidism. AB - BACKGROUND: Autoimmune diseases are characterized by induced parenchymal expression of major histocompatibility complex (MHC) class II antigens and circulating autoantibodies directed toward surface structures on the target cells. MHC class II expressions can be modified by viral infections of potential pathogenic importance in autoimmune reactions. Primary hyperparathyroidism exhibits incompletely clarified cause. METHODS: With cryosections, human parathyroid glands were stained with monoclonal antibodies to MHC class II antigens according to a peroxidase-antiperoxidase technique. Human parathyroid adenoma tissue transplanted to nude mice and rat parathyroid glands was tested with serum from patients with hyperparathyroidism and control subjects. RESULTS: Induced MHC class II expression was demonstrated on parathyroid parenchymal cells in 13 of 54 adenomatous and eight of 23 hyperplastic glands of patients with primary hyperparathyroidism. This reactivity was absent in 12 normal glands, nine normal-sized glands associated with the adenomas, and 17 enlarged glands of patients with hyperparathyroidism caused by uremia. Staining of parathyroid tissue was found with serum from 27 of 38 patients with primary hyperparathyroidism, whereas this reactivity was absent on rat thyroid and pancreatic tissue, as well as with control sera. CONCLUSION: The concurrent induction of MHC class II antigen expression and c circulating antiparathyroid autoantibodies in 16 or 38 patients with primary hyperparathyroidism suggests hitherto unrecognized immunologic involvement in this disease. PMID- 9736903 TI - Pancreatic invasion as the prognostic indicator of duodenal adenocarcinoma treated by pancreatoduodenectomy plus extended lymphadenectomy. AB - Pancreatoduodenectomy has become the standard procedure in resection of the duodenal adenocarcinoma, and some adjuvant therapies can be added to obtain further improvement in postoperative outcome. However, for patient selection, it is necessary to have a predictive indicator showing, if possible before laparotomy, which instances are noncurable by surgery alone or need adjuvant therapies. METHODS: A retrospective analysis was made for 24 consecutive patients whose duodenal adenocarcinoma were treated by pancreatoduodenectomy plus a wide range of lymphadenectomies without any adjuvant therapies at Osaka Medical Center for Cancer and Cardiovascular Diseases. Patient survival rates were related to macroscopic and microscopic findings and to findings obtained by preoperative imaging techniques. RESULTS: The overall survival rate was 69% at 3 years and 57% at 5 years; locoregional recurrence was the primary cause of death. Although the 5-year survival rate was 44% in patients with nodal involvement and 76% in those without, this difference did not reach statistical significance (P = .079). Instead, invasion into the pancreatic parenchyma at a macroscopic level was the most significant prognostic factor; the 5-year survival rate was 78% in the 16 patients without and 16% in the 8 patients with pancreatic invasion (P = .0047). Invasion into the pancreas correlated well with the angiographic findings; the 5 year survival rate was 25% in patients whose angiograms delineated the pancreatic invasion and 83% in patients whose angiograms did not (P = .0084). CONCLUSION: When duodenal adenocarcinoma was treated by pancreatoduodenectomy plus a wide range of lymphadenectomy, pancreatic invasion at a macroscopic level was most associated with patient survival. Pancreatic invasion was well delineated by the preoperative angiogram, which would be helpful in patient selection. PMID- 9736904 TI - The effect of laparotomy and laparoscopy on the establishment of spontaneous tumor metastases. AB - BACKGROUND: Surgical extirpation of solid tumors may be entirely possible, and the consequence of surgical excision is invariably the release of tumor cells into the systemic circulation. The aim of this study was to determine whether laparotomy affects the establishment of spontaneous pulmonary metastases after excision of the primary tumor in a murine flank tumor model and to determine possible underlying immune abnormalities. METHODS: An initial experiment was carried out to compare the development of gross spontaneous pulmonary metastases in the presence of a primary flank tumor and after excision of the tumor in C57/BL6 female mice. Another group of mice had flank tumors excised and were simultaneously randomized to undergo anesthetic only (control), laparoscopy, or laparotomy, after which the subsequent development of pulmonary metastases was determined. Finally, a third experiment entailed determination of natural killer cell (NK) cytotoxicity and the effect of splenic macrophages on NK cytotoxicity at days 1,7, and 14 after tumor excision. RESULTS: Excision of the primary tumor resulted in a significant increase in the number of pulmonary metastases in mice compared with mice that did not have tumors excised (P = .01). Both laparotomy and laparoscopy significantly increased the number of spontaneous pulmonary metastases after tumor excision compared with controls (P < or = .01), and there was also a significant difference between laparotomy and laparoscopy groups (P = .00). NK cytotoxicity was significantly suppressed at all time points after operation in the laparotomy group compared with both the laparoscopy group and the controls (P < or = .01). Suppression occurred after laparoscopy at 24 hours after the procedure compared with controls (P = .00); by day 7 this difference was not significant, but as day 14 there was again a significant suppression (P < or = .03). Splenic macrophages appeared to be a suppressor to natural killer cell cytotoxicity (NKCC) in the corresponding groups and at the corresponding time points. CONCLUSIONS: The differential establishment of spontaneous metastases after tumor excision and laparotomy and, to a lesser extent, laparoscopy results in lowered host antitumor surveillance and may be mediated at least in part by the generation of splenic suppressor cells in the early postoperative period, causing a more marked and prolonged effect after laparotomy than after laparoscopy. PMID- 9736905 TI - Staged hepatectomy after emergency transcatheter arterial embolization for ruptured hepatocellular carcinoma. AB - BACKGROUND: Staged hepatectomy after emergency transcatheter arterial embolization (TAE) has been advocated in ruptured hepatocellular carcinoma (HCC). However, there have been no reports of clinical series of this strategy. The purpose of this study was to evaluate the protocol of this therapeutic strategy. METHODS: Sixteen patients with suspected rupture of HCC were included in the study. After emergency TAE, tumor resectability was assessed, followed by staged hepatectomy or repeated TAE. The patients were reevaluated with regard to rupture of HCCs. RESULTS: Primary hemostasis was achieved successfully in all patients. Eleven patients were finally judged to have experienced HCC rupture. Seven of them underwent staged hepatectomy; the other four underwent repeated TAE because their tumors were considered unresectable. Survival time tended to be longer, but not to a significant extent, in patients who underwent hepatectomy (range, 139 to 1527 days; median, 375 days) than in those treated by TAE alone (range, 43 to 1317 days; median, 158 days). CONCLUSIONS: Staged hepatectomy after TAE is a rational treatment for patients with ruptured HCC. Although TAE is highly effective for initial hemostasis, hepatectomy appears to provide better long-term survival. PMID- 9736907 TI - Treatment of hepatic metastases of colorectal cancer by electrochemotherapy: an experimental study in the rat. AB - BACKGROUND: Electrochemotherapy, which consist of local or systemic administration of a cytotoxic agent followed by application of electric pulses to a tumor, has proved effective for various types of tumors in animals and for cutaneous and head and neck cancers in human beings. This is the first study to investigate the efficacy of electrochemotherapy for treatment of hepatic metastases of colorectal cancer in the rat. METHODS: After induction of a solitary hepatic metastasis in 36 male BDIX rats, the animals were randomized to one of four groups: B-E-(no treatment), B+E-(intratumoral bleomycin), B-E+ (application of electric pulses to the tumor), and B+E+ (electrochemotherapy: intratumoral bleomycin followed by application of electric pulses). RESULTS: Groups B-E and B-E+ had no tumor response. Group B+E had one partial response. Group B+E+ had seven partial responses and two complete responses. The difference in terms of response between group B+E+ and the other three groups was statistically significant (P < .05). Comparison of the mean posttherapy tumor volumes (B-E-, 50.6 mm3; B+E-, 58.7 mm3; B-E+, 46 mm3; and B+E+, 5.65 mm3) revealed a significantly smaller residual tumor in group B+E+ than in the other three groups (P < .05). CONCLUSIONS: Electrochemotherapy is an effective means to reduce the volume of hepatic metastases of colorectal cancer in the rat. Additional research is required to determine the optimum treatment duration, dose effects, volume of tumor that can be treated by electrochemotherapy, and impact on survival. Such experimental studies are indispensable prerequisites for clinical trials. PMID- 9736908 TI - Phenylacetate inhibits isoprenoid biosynthesis and suppresses growth of human pancreatic carcinoma. AB - BACKGROUND: Phenylacetate is growth inhibitor for a variety of tumors at concentration that have been safely achieved in human beings. This antitumor effect is related to inhibition of the isoprenoid synthetic pathway by blocking the enzyme, mevalonate pyrophosphate (MVAPP) decarboxylase. The purpose of this study was to evaluate the effects of phenylacetate on human pancreatic carcinoma. METHODS: For in vitro studies, six human pancreatic carcinoma cell lines (BxPc, AsPc, MIAPaCa-2 Panc-1, CFPAc, and HS 766T) were studied. For in vivo studies, nude mice were inoculated with pancreatic cells (BxPc and MIA PaCa-2) and randomized to receive phenylacetate or saline control. RESULTS: Phenylacetate produces reversible in vitro growth arrest at doses of 2.5 to 10 mmol. The antiproliferative effect is cytostatic, producing accumulation of cells in G1, and is not associated with cell toxicity. Systemic treatment of nude mice bearing heterotopic human pancreatic carcinoma results in growth inhibition of tumors without host toxicity. Phenylacetate blocks the processing of mevalonate to isopentenyl-pyrophosphate by inhibiting MVAPP and exhibits suppression of biosynthetic pathways requiring isoprenoids, including cholesterol and dolichol biosynthesis, protein glycosylation, and isoprenylation of proteins. CONCLUSIONS: These results indicate that phenylacetate has cytostatic activity in pancreatic carcinoma and support the conclusion that suppression of multiple biosynthetic pathways requiring isoprenoids is contributing to the drug's antiproliferative action. The safety profile and efficacy of phenylacetate make it an attractive agent for the treatment of pancreatic cancer. PMID- 9736909 TI - Immunoaffinity localization of the enzyme xanthine oxidase on the outside surface of the endothelial cell plasma membrane. AB - BACKGROUND: Reactive oxygen metabolites generated from endothelial xanthine oxidase (XO) trigger reperfusion injury in many organs. We evaluated the possibility that endothelial XO was localized on the endothelial cell surface, as well as within the cytoplasm. METHODS: Primary cultures of bovine (BAECs) and porcine (PAECs) aortic endothelial cells were grown in media documented to be free of XO. Polyclonal and monoclonal antibodies were developed against XO. These antibodies were used to evaluate BAEC and PAEC for cell surface XO through immunofluorescence staining, hybridoma cell surface labeling, and endothelial cell surface binding. RESULTS: These antibodies bound specifically to the surface of these cells when the membrane was shown to be intact and impermeable (and the cytoplasm inaccessible) to immunoglobulins Moreover, hybridoma cells expressing monoclonal antibody to XO bound specifically to the endothelial cell surface. Finally, intact endothelial cells bound specifically to the anti-XO polyclonal antibodies immobilized to the surface of a Petri dish. The integrity of these endothelial cell plasma membranes was demonstrated by the subsequent growth and replication of these cells in culture. CONCLUSIONS: These findings indicate that XO is present on the outside surface of the endothelial cell plasma membrane. This would not only explain the known in vivo efficacy of intravascularly administered large molecular weight antioxidants (such as superoxide dismutase) but could have important implications for inflammatory signaling. PMID- 9736910 TI - Stenting does not decompress the pancreatic duct as effectively as surgery in experimental chronic pancreatitis. AB - BACKGROUND: In humans with chronic pancreatitis (CP), pancreatic interstitial pressure (IP) is elevated and pancreatic blood flow (PBF) is reduced. The efficacy of surgical decompression (SD) of the pancreatic duct (ie, pancreaticojejunostomy) is believed to be due to its ability to decrease IP and pancreatic vascular resistance (Rp), which increases PBF. Pancreatic duct stenting (STE) also probably reduces IP and Rp, which may explain its efficacy. The purpose of this study was to compare the efficacy of SD with STE. METHODS: CP in cats was created by narrowing the main pancreatic duct. Six weeks later, CP and normal pancreata were isolated and perfused ex vivo under basal conditions and after secretin stimulation. In normal and CP glands, IP and perfusion pressure were measured and Rp (U) was calculated. In two additional groups, the pancreatic duct was decompressed, either by stenting or by complete transection of the duct with a longitudinal capsulotomy. RESULTS: In CP glands, IP and Rp were increased and secretory output was markedly reduced compared with the normal (0.65 +/- 0.30 mm Hg and 0.46 +/- 0.04 U vs 3.90 +/- 0.80 mm Hg and 1.68 +/- 0.05 U; P < .05). Secretin administration (2 units) increased IP and Rp in CP glands (6.60 +/- 1.10 mm Hg and 2.87 +/- 0.07 U; P < .05), but these values did not chang in normal glands (0.81 +/- 0.20 and 0.53 +/- 0.03 U; NS). STE and SD decreased IP and Rp in CP glands (2.20 +/- 0.20 to 1.0 +/- 0.40 mm Hg and 1.20 +/ 0.015 to 0.90 +/- 0.01 U, respectively; P < .05). Both methods prevented an increase of IP and Rp after secretin administration. IP and Rp decreased to a greater degree following SD, compared with STE (P < .05). CONCLUSIONS: Both STE and SD decreased IP and Rp in this experimental model of CP. However, SD was significantly more effective than STE. PMID- 9736911 TI - Early isotonic saline resuscitation from uncontrolled hemorrhage in rats. AB - BACKGROUND: Attempts to modify traditional fluid resuscitation have been based on animal models that evaluate several variables including anesthesia. This study presents the effects of early saline resuscitation from severe uncontrolled hemorrhage unanesthetized rats. METHODS: Sixty-three female Sprague-Dawley rats were equally divided into three groups: group A, nonresuscitated; and groups B and C, resuscitated ;with isotonic saline (40 and 80 mL/kg, respectively). Hemodynamics, blood loss, survival time, and mortality were recorded for 360 minutes after the hemorrhage, which was initiated by 75% resection of the tail. RESULTS: In group C, 80 mL/kg of saline significantly lowered mortality (24% vs 76% and 71% for groups A and B, respectively) with concomitant increases in mean survival time (241 +/- 103 min vs 146 +/- 108 and 175 +/- 92 min for groups A and B, respectively). There were no statistically significant differences in blood loss, hematocrit, or hemodynamic parameters among the groups. CONCLUSIONS: Early and adequate isotonic saline resuscitation of unanesthetized rats improved outcome despite continuing hemorrhage. The significantly lower mortality rate and increased survival time were not a result of transiently improved arterial pressure and did not correlate with blood loss. No significant bleeding increases were noted in the resuscitated groups. PMID- 9736912 TI - Prognosis of hepatocellular carcinoma in relation to treatment: a multivariate analysis of 178 patients from a single European institution. AB - BACKGROUND: Because the prognosis of patients with hepatocellular carcinoma is not fully understood, particularly regarding therapy, we have evaluated it in a series of patients with a homogeneous diagnostic and therapeutic work-up. METHODS: From 1985 to 1996, 42 variables were recorded prospectively in 178 constructive patients who had a diagnosis of hepatocellular carcinoma. Treatment consisted of liver transplantation ( n = 22), partial hepatectomy (n = 11), arterial, chemoembolization ( n = 52), systemic or regional chemotherapy (n = 51), and other therapies (n = 5); 37 patients received no specific therapy. Statistical analysis was performed according to a Cox model. RESULTS: There were no differences between the survival of patients receiving chemotherapy, other therapies, or no treatment (control group n = 93). survival rates a 1,3, and 5 years were 81%, 74%, and 74% for liver transplantation, 72%, 58%, and 58% for hepatectomy, 55%, 26%, and 13% for chemoembolization, and 13%, 3%, and 0% for the control group. Cirrhosis, systemic syndrome, bilobar involvement, Child's stage C disease, and treatment were independent predictors of survival. CONCLUSIONS: This series shows that certain easily accessible parameters may help establish individual prognosis and stratify patients in clinical trials and indicates that chemoembolization, partial resection, and liver transplantation can prolong life expectancy of patients with hepatocellular carcinoma. PMID- 9736914 TI - Effects of giving "high-dose" insulin in burn patients. PMID- 9736913 TI - IT 9302, a synthetic interleukin-10 agonist, diminishes acute lung injury in rabbits with acute necrotizing pancreatitis. AB - BACKGROUND: Proinflammatory cytokines (eg, tumor necrosis factor [TNF]-alpha, interleukin [IL]-1 and Il- 8) are believed to play an important role in the pathogenesis of acute necrotizing pancreatitis (ANP) and its systemic complications. Recently, IL-10 has emerged as a major anti-inflammatory cytokine, inhibiting the secretion and activities of inflammatory cytokines. Further, a protective effect of IL-10 has recently been shown in experimental acute pancreatitis. The purpose of this study was to test the potential role of a newly developed IL-10 agonist, IT 9302, in a model of ANP in rabbits. METHODS: ANP was induced in 18 rabbits by retrograde injection of 5% chenodeoxycholic acid in the pancreatic duct, followed by duct ligation. The rabbits were allocated to pretreatment with intravenous physiologic saline solution or IT 9302 (200 micrograms/kg) 30 minutes before the induction of ANP. RESULTS: Injection of IT 9302 resulted in a significant reduction in the blood levels of TNF-alpha and IL 8 from 3 to 6 hours. IT 9302 also reduced the amount of ascitic fluid and significantly inhibited neutrophil infiltration and margination, as well as the number of CD11b- and CD18-positive cells in the lung tissues. By contrast, the local pancreatic necrosis, as well as the biochemical changes such as serum amylase, lipase, and calcium, was sever and similar in both groups. Survival was improved significantly after treatment with IT 9302. CONCLUSIONS: As expected, IT 9302 cannot change the degree of ANP induced by 5% bile acid but does reduce mortality rates and the development of acute lung injury, probably through the inhibition of circulating levels of TNF-alpha, IL-8, and the expression of the adhesion molecule complex CD11b/CD18. PMID- 9736915 TI - Next generation therapeutics. PMID- 9736916 TI - Peptidomimetic design. AB - Major discoveries have been made of new type-I and type-III peptidomimetic inhibitors of peptide-derived systems. Innovative reversible inhibitors of cysteine proteases and renin, and additional examples of peptidomimetic inhibitors of interleukin-1 beta-converting enzyme, neutral endopeptidase, herpes simplex virus protease, thrombin, HIV protease, Ras farnesyltransferase, the RGD motif, Factor Xa and various aspartic proteases have been discovered. PMID- 9736917 TI - Very late antigen 4 (VLA4) antagonists as anti-inflammatory agents. AB - In recent years antagonists of very late antigen-4 (VLA4, also known as integrin alpha(4) beta(1)) have shown great promise in treating inflammatory disorders in a number of animal models. The most advanced in this endeavor is a humanized anti alpha(4) antibody, Antegren, which is in phase II clinical trials for multiple sclerosis. The first reported small-molecule VLA4 antagonist to advance into clinical trials is currently in phase I as an aerosol for treating asthma. A number of peptides, cyclic peptides and peptidomimetics have been disclosed and are in preliminary stages of development. PMID- 9736918 TI - Cardiovascular chemotherapy: anticoagulants. AB - Anticoagulant therapy has changed dramatically during the past two years. Low molecular weight heparin has substantially replaced unfractionated heparin as the parenteral anticoagulant of choice. The use of warfarin has substantially increased, especially for prevention of stroke in the setting of atrial fibrillation. It has become clear that warfarin cannot be administered effectively in an unmonitored fixed-dose fashion. The parenteral direct thrombin inhibitor desirudin was shown to be efficacious in the prevention of deep vein thrombosis in man. Small thrombin and factor Xa inhibitors with in vivo oral anticoagulant activity have been identified. PMID- 9736919 TI - Matrix metalloproteinases. AB - Matrix metalloproteinases are a family of highly regulated peptidases that are collectively responsible for the degradation of extracellular matrix during tissue remodeling. Dysregulated activity has long been implicated in the pathologies of cancer and arthritis, and the number of diseases more recently associated with these enzymes has been increasing. In the past year, new transgenic models of matrix metalloproteinase knockouts have been described, allowing the direct assessment of specific enzyme activity in particular disease models. In addition, more selective inhibitors with improved pharmacokinetic profiles have entered clinical trials, allowing the assessment of the safety and efficacy of this strategy. PMID- 9736920 TI - Chemotherapeutic potential of phosphodiesterase inhibitors. AB - The application of molecular cloning has revealed the phenomenal diversity and complexity of the phosphodiesterase isoenzyme family. Thus, more than 30 human phosphodiesterases are now known; all are apparently necessary for the seemingly simple task of hydrolysing the 3'-ester bond of either cyclic adenosine monophosphate or cyclic guanosine monophosphate. The availability of phosphodiesterase isoenzymes as pure recombinant proteins has greatly facilitated the identification of potent, selective inhibitors. The potential of these inhibitors to therapeutically exploit the molecular diversity of the phosphodiesterases has progressed significantly. A number of drugs are in clinical trials for asthma, and Viagra has become the first selective phosphodiesterase inhibitor to be approved by the US Food and Drug Administration. PMID- 9736921 TI - Design of selective inhibitors of cyclooxygenase-2 as nonulcerogenic anti inflammatory agents. AB - The discovery of a second isoform of cyclooxygenase (cyclooxygenase-2) that is expressed in inflammatory cells and the central nervous system, but not in the gastric mucosa, offers the possibility of developing anti-inflammatory and analgesic agents that lack the gastrointestinal side effects of currently available nonsteroidal anti-inflammatory drugs. Lead compounds from several different structural classes have been identified and shown to be slow, tight binding inhibitors that express their selectivity for cyclooxygenase-2 in the time-dependent step. The determination of structures of enzyme-inhibitor co crystals along with site-directed mutagenesis experiments reveal the molecular basis for selectivity of some, but not all, inhibitors. Preclinical and clinical studies suggest cyclooxygenase-2 inhibitors are highly promising new agents for the treatment of pain and inflammation, and for the prevention of cancer. PMID- 9736922 TI - Design of isoform-selective inhibitors of nitric oxide synthase. AB - Nitric oxide synthase, the mammalian enzyme catalyzing the oxidation of L arginine to L-citrulline and nitric oxide, is present in three isoforms that have distinct physiological roles. Overstimulation or overexpression of individual nitric oxide synthase isoforms plays a role in a wide range of disorders including septic shock, arthritis, diabetes, ischemia-reperfusion injury, pain and various neurodegenerative diseases. Animal studies and early clinical trials suggest that nitric oxide synthase inhibitors could be therapeutic in many of these disorders, but preservation of physiologically important nitric oxide synthase functions might require use of isoform-selective inhibitors. Within the past few years both amino acid and nonamino acid nitric oxide synthase inhibitors with pharmacologically useful isoform selectivity have been reported. Selectivity has been achieved on the basis of initial binding affinity and, for mechanism based inactivators, on the basis of isoform-dependent catalytic activation; particularly interesting are N5-(1-imino-3-butenyl)-L-ornithine, ARL 17477, 1400W and S-(2-aminoethyl)isothiourea. PMID- 9736923 TI - Therapeutic potential of selective modulators of nuclear receptor action. AB - Nuclear receptors belong to a superfamily of ligand-inducible transcription factors that, in addition to directly regulating their cognate gene programs, can also mutually interfere with other signaling pathways. The recent identification of selective agonists/antagonists of the glucocorticoid, retinoid and estrogen receptors suggests that it might be possible to selectively elicit only a subset of the nuclear receptor functions that are induced by the natural ligand, with the aim of increasing the functional and, perhaps, tissue selectivity of nuclear receptor ligands and reducing unwanted side effects. PMID- 9736924 TI - Therapeutic potential of selective estrogen receptor modulators. AB - The hormone estradiol has effects on many tissues in both males and females. Some of these effects, such as inhibition of cancer growth and modulation of the devastating effects of aging on bone, brain, skin and bladder, are good. Others, such as the effect on the breast and endometrium, are undesirable. The task of designing drugs that would have only the good effects of estradiol has, until recently, seemed almost impossible because it was thought that there was only one estrogen receptor and that an estrogenic agent was definitively categorized as an estrogen agonist or an antagonist. More recently we have learnt that there are two estrogen receptors which, under certain conditions, have opposite effects on gene transcription. In addition, it is now understood that agents cannot be described as agonists or antagonists because a single agent can be an agonist in one tissue and an antagonist in another. The term 'selective' estrogen receptor modulator' was designed to incorporate this. The idea of estrogen receptor modulators has raised new hope that tissue specific estrogens or anti-estrogens can be designed. PMID- 9736925 TI - Inducible control of gene expression: prospects for gene therapy. AB - A number of drug-related gene expression systems are available for controlling target gene transcription through the use of small-molecule inducing compounds. While the utility of such systems has been demonstrated in vitro and in transgenic mice, recent improvements are likely to make these systems more amenable for use in a therapeutic context, such as gene therapy. These improvements include further optimization of the antiprogestin-regulated gene switch, rendering it more sensitive to RU486, and the synthesis of nonimmunosuppressive rapamycin analogs for use in dimerization-based strategies of gene regulation. PMID- 9736926 TI - Antisense therapeutics. AB - The field of antisense therapeutics has attracted great interest during the past decade. A large body of literature has recently appeared in which the antisense mechanism is claimed to be involved and a number of human clinical trials are underway. Questions regarding the specificity of action and side effects of antisense phosphorothioate oligonucleotides have arisen simultaneously. PMID- 9736927 TI - Novel approaches to the discovery of antimicrobial agents. AB - With the completion of numerous bacterial genome sequences, the discovery of antibacterial drugs has fully entered the genomic era. The strategies for effectively using genomic information for target identification, target characterization, screen development and compound evaluation are emerging, and have greatly increased the number of antibacterial targets available for screening. Fortunately, simultaneous efforts in improving miniaturization, robotics and database tools are underway so that the potential of genomics can be realized. PMID- 9736928 TI - The discovery of potent and selective dopamine D4 receptor antagonists. AB - The identification of a novel dopamine receptor subtype, referred to as the D4 receptor, which binds the atypical antipsychotic drug clozapine with high potency, has led to the initiation of a drug discovery program that aims to find novel inhibitors of this receptor subtype. A selective screening strategy was utilized, in which 4500 compounds chosen on the basis of structural similarities to known biogenic amine receptor antagonists were tested against both the D4 and D2 dopamine receptor subtypes. A potent D4-selective compound was discovered. PMID- 9736929 TI - Rational treatment of addiction. AB - Treatment of alcohol and drug addictions, which has been neglected medically for a long time, is currently sparked with optimism. Craving for alcohol can be treated with two newly registered drugs: naltrexone and acamprosate. New approaches to symptom relief during detoxification or during maintenance therapies are rationally based on experimental and clinical work. It is now clear that addictive drugs are surrogates of natural substances involved in the 'reward system'. PMID- 9736930 TI - Encapsulation of proteins for improved delivery. AB - Two recent advances have enabled the development of clinically useful, injectable, sustained-release protein formulations. The first is a nonaqueous, cryogenic atomization process that encapsulates the protein into microspheres composed of a biodegradable polymer from which the protein is released slowly. The second consists of numerous ways of maintaining protein stability within the microspheres for extended periods after injection. In addition to allowing less frequent administration of protein drugs, at possibly lower overall doses, it is possible that sustained-release formulations will justify commercial development of proteins that could not be marketed as solution formulations. PMID- 9736931 TI - [Gustav Killian--a pioneer in endoscopy]. PMID- 9736932 TI - [Current aspects in the design of endoscopes]. PMID- 9736933 TI - [Percutaneous endoscopic gastrostomy (PEG) tube: consultation field for the ENT physician?]. PMID- 9736934 TI - [Argon plasma surgery. Technology and possible application to ENT medicine]. PMID- 9736935 TI - [Retrograde and fiber optic intubation successfully combined]. PMID- 9736936 TI - [Tracheobronchoscopy and esophagoscopy in otorhinolaryngology. An assessment of current status]. AB - Its is the task of each medical specialty to develop guidelines for diagnosis and therapies. Examinations done by several specialties should follow a common consensus. A randomized survey at 70 German ENT departments investigated the current position of tracheobronchoscopy and esophagoscopy at each institution. Sixty questionnaires were evaluable. Altogether 8,295 tracheobronchoscopies and 10,404 esophagoscopies were performed. Thirty-six percent of all tracheobronchoscopies and 6% of all esophagoscopies were done with a flexible system. Approximately 58% of all tracheobronchoscopies and 55% of all esophagoscopies were performed for tumor staging. Complications during tracheobronchoscopy occurred in 0.8% of cases and in 0.58% of the esophagoscopies. Using these data an interdisciplinary quality assurance concept was developed for tracheobronchoscopy and esophagoscopy. Current experience has shown that a otolaryngologists in Germany mainly perform rigid tracheobronchoscopy and esophagoscopy. Although endoscopy is mostly done in cases with varied anatomic structures, complications are very rare and comparable to flexible techniques. Greater experience with flexible systems also is to be encouraged in ENT departments. PMID- 9736937 TI - [Esophagoscopy guideline. Guidelines of the German Society of Otorhinolaryngology, Head and Neck surgery]. PMID- 9736938 TI - [Tracheobronchoscopy guideline. Guidelines of the German Society of Otorhinolaryngology, head and neck surgery]. PMID- 9736939 TI - [Flexible endoscopy in the ENT area]. AB - Although rigid esophagoscopy and tracheobronchoscopy have always been a domain of the ENT surgeon, the development of flexible endoscopes has increased diagnostic and therapeutic indications in clinical practice. Component elements of fiberendoscopes and CCD endoscopes are shown. Techniques in flexible bronchoscopy, flexible esophagoscopy and rhinolaryngoscopy are explained. General and special indications of fiberendoscopy are listed. Possibilities and limitations of flexible bronchoscopy are shown. Flexible techniques for removing foreign bodies of the esophagus and indications for mini-endoscopy are demonstrated. Experience has shown that flexible endoscopy has brought the diagnosis and therapy of tubular organs into a new perspective. Since flexible techniques do not replace rigid ones and complement each another, ENT surgeons must be trained in both techniques. PMID- 9736940 TI - [Long-term experience with percutaneous endoscopic controlled gastrostomy (PEG) in ENT tumor patients]. AB - Percutaneous endoscopically controlled gastrostomy (PEG) enables patients suffering from a tumor of the upper aerodigestive tract to receive direct gastric feedings. The procedure also avoids the social stigma of a nasal feeding tube. The results of 630 PEG procedures used in 555 patients suffering from various head and neck cancers are reported. The mean age of the patients was 58.0 years with a range from 11 to 92 years. The PEG procedure was carried out under local anesthesia in 60% of the cases and with general anesthesia in 40%. In 512 patients the initial PEG procedure was successful while 43 of the patients require a second PEG procedure after loss of the PEG. In 19 patients the PEG procedure was not successful because of tumor obstruction or it was not possible to perform endoscopy. Twenty-four patients were successfully treated in a second or third session. Although 97% (n = 563) of all 555 patients and 92% (n = 579) of all PEG procedures were successful. The mean duration of PEG use was 243 (range: 0-2271 days). In 66 patients (10.5%) complications occurred but severe complications developed in only 8 patients (1.3%). Operative interventions were necessary in two cases. No deaths resulted from the PEG. These findings show that the PEG technique is safe to do with only few complications when performed by a skilled team. PMID- 9736941 TI - [Problematic intubation in the patient with laryngeal-hypopharyngeal carcinoma. Retrograde controlled fiber bronchoscopy technique]. AB - Retrograde or fiberoptic intubation techniques are recommended for patients in whom intubation is difficult; however, each method has its own limitations. Good results have been reported with a combination of both techniques, i.e. retrograde passage of a guidewire through the cricothyroid membrane to guide a fiberoptic bronchoscope. The practicality, success and complication rates of our retrograde guided fiberoptic bronchoscopic technique (RGFT) were studied prospectively in 93 patients with obstructing tumors scheduled for laryngectomy. The techniques showed itself to be successful, practical and safe, with negligible complications in 89/93 patients (96%). The ability to insert the bronchoscope by means of a guidewire and to direct the intubation procedure optically was found to be advantageous. Limitations with extreme obesity and in two other patients with advanced obstructive carcinomas of the larynx. Additionally, use of the tracheal puncture allows the RGFT to be integrated into clinical medical education as a preparatory exercise for emergency coniotomy. PMID- 9736942 TI - [Argon plasma surgery (APC) in the upper aerodigestive tract. Initial results]. AB - Cold knife surgery, electrosurgery and laser surgery all offer techniques, instruments, equipment and systems for resecting and destroying mucosal lesions and for hemostasis in the upper aerodigestive tract. When used in the head and neck, argon plasma surgery (APS) offers a new, contact-free, electrosurgical technique in which high frequency current is applied through ionized, and thus electrically conductive, argon(argon plasma) to the tissue undergoing treatment. Especially noteworthy in APS are its advantages for removing a lesion and controlling bleeding: the technique is easy to control, and the depth of the thermal tissue destruction is limited to a maximum of 3 mm even in wide-area application, so that damage to adjacent or submucosal tissues can be avoided. Initial results with APS in the reduction of hyperplastic nasal turbinates, treatment of hereditary hemorrhagic teleangiectasia (Osler's disease) in the nasal mucosa, and in treating progressive juvenile papillomatosis of the larynx have shown clear advantages for APS over other methods used. PMID- 9736944 TI - [Objective description of voice quality using the hoarseness diagram]. AB - The need for an objective assessment of voice quality can be seen in the increasing use of acoustic analysis methods for clinical diagnosis and research. The hoarseness diagram allows clinicians to objectively describe even highly disturbed or aphonic votes. Its application possibilities are illustrated in three case studies in which changes in voice quality were monitored during voice rehabilitation. The distributions of voice groups that were defined on the basis of specific pathophysiological phonation conditions were then compared for their acoustic differences. The groups comprised various phonation conditions after the resection of laryngeal tumors and different types of laryngeal paralyses. Interpretation of the results suggests a direct correspondence of the hoarseness diagram coordinates and the irregularity of vibration on the one side and the degree of glottal closure on the other. This illustrates the potential usefulness of the hoarseness diagram in a clinical context. PMID- 9736943 TI - [Ciaglia percutaneous dilatation tracheotomy with endoscopic control. Analysis of complication-fraught steps]. AB - Since the first description by Ciaglia in 1985, percutaneous dilatational tracheostomy (PDT) has emerged worldwide to be a favored and frequently used technique for airway control. Advantages are the rapidity, simplicity and independence of the physician. These characteristics tempt clinicians to perform the procedure uncritically and thus produce complications. Based on our experiences with the technique and on a critical analysis of the literature the indications and optimal operative technique are discussed. Operative steps are illustrated and potential pitfalls (anatomic variations, injury to vital structures and bleeding) are defined. In our experience endoscopic control plays a crucial role in preventing intraoperative complications. ENT colleagues should know the technique because of their experience with tracheotomized patients and should offer their services to other disciplines. PMID- 9736945 TI - [Cystic tumor of the nose root. Acquired epidermoid cyst]. PMID- 9736946 TI - [Tullio phenomenon after cochlear implantation]. AB - The Tullio phenomenon is defined as an acoustically inducible vestibular disorder that was first described in 1929. In an animal experiment Tullio provoked acoustic oscillations at the labyrinth followed by signs of imbalance. In the literature this phenomenon can be found in healthy but sensitive persons as well as in patients with Meniere's disease and patients with lesions between the stapes, footplate and the membranous labyrinth caused by fractures, stapes dislocations, labyrinthitis or perilymphatic fistulas. In this case report a patient complained about vertigo after cochlear implantation provoked by acoustical stimulation at a sound pressure level above 90 dB independent of the cochlear implant (CI). During tympanoscopy we found scar tissue surrounding the ossicles after CI. After disconnecting the ossicular chain no vertigo or nystagmus could be provoked. After CI regular ENT examinations and appropriate explorations of postoperative complaints are necessary. Vertigo especially requires very careful diagnostic procedures. PMID- 9736947 TI - [Hygiene in ENT. What is necessary, what is possible?]. PMID- 9736948 TI - XII Panamerican Congress of Rheumatology. Montreal, Canada. June 21-25, 1998. Abstracts. PMID- 9736949 TI - Neuroreceptor mapping 98. The 2nd International Symposium on Functional Neuroreceptor Mapping of Living Brain. Ann Arbor, Michigan, USA. June 12-14 1998. Abstracts. PMID- 9736950 TI - [The evaluation of patients with ischemic cerebral lesions by CT, SPECT and qEEG in acute, subacute and chronic phases]. AB - INTRODUCTION: SPECT, EEG AND CT scan offer information with several pathophysiologic meanings. Their results vary with time and according to the vascular affected territory. OBJECTIVE: We wanted to study how the sensibility varies and the relationship with the clinic of SPECT, qEEG and CT scan in the acute, subacute and chronic stages and according to the vascular affected territory. We also wanted to analyze the several pathophysiologic aspects of the cerebral ischemia. METHODS: Thirty-six patients with symptoms of hemispheric stroke were evaluated with CT scan, qEEG, SPECT99mTc-HMPAO during the acute (0-5 days), subacute (0-15 days) and chronic (16 days to 1 year) stages. RESULTS: The decrease of ipsilateral CBF depend on the time (p = 0.0061), being not very frequent during the two first weeks. The qEEG was the most sensitive study in the first phase, its sensibility did not depend on the vascular affected territory and was dependent on the time (p = 0.0011), diminishing in the chronic phase. The slow activity was habitually ipsilateral. The CT scan was the less sensitive study. CONCLUSION: After 24 hours and until the second week, there is habitually an increase of the ipsilateral rCBF. The luxury perfusion could explain the fogging effect in the CT scan. The slow activity of the qEEG represents the alteration of the oxygen metabolism. The interpretation of the variation of the CBF and the qEEG allow us to define oligemia of the ischemia and between reactive hyperemia and the increase of CBF due to the necrotic tissue. PMID- 9736951 TI - [The involvement of the parenchyma of the central nervous system in Behcet disease]. AB - INTRODUCTION: Behcet disease is a systemic form of vasculitis which presents with neurological symptoms with a frequency varying between 16 and 40%. Involvement of the parenchyma has been found to worsen the prognosis in patients with neuroBehcet (NB). OBJECTIVE: To review the clinical features and course of patients with NB involving the parenchyma of the central nervous system (CNS). CLINICAL CASES: Seven patients with Behcet disease and neurological localizing signs were seen in our hospital between 1989 and 1996. The initial diagnosis was of ischemic ictus in five of the seven patients. Both neuroimaging studies and investigation of the cerebrospinal fluid were always pathological in all cases. Vascular studies (arteriography and echo-Doppler of the supra-aortic trunks) were normal. One patient died. Four patients had serious sequelae following treatment. CONCLUSION: NB should be included in the different diagnosis of ictus. Involvement of the parenchyma of the CNS was accompanied by lymphocytic meningitis, perhaps also leading to a worse functional prognosis. PMID- 9736952 TI - [Chiari malformations and hydrosyringomyelia]. AB - INTRODUCTION: The association of hindbrain herniation, better known as Chiari malformation, and cystic cavitation of the spinal cord or hydrosyringomyelia has been well described in the literature although there is little consensus regarding its etiology, pathophysiology or optimal treatment. DEVELOPMENT: Despite a well-accepted and utilized classification system of both Chiari malformation and hydrosyringomyelia, there remains considerable disagreement regarding the proper management of these patients. CONCLUSIONS: This article reviews several popular theories on the etiology and pathophysiology of this disorder, briefly discusses the clinical features and radiologic findings associated with Chiari malformation and hydrosyringomyelia, and reviews basic surgical techniques for decompression of the cranial-cervical junction and treatment of the hydrosyringomyelia. PMID- 9736953 TI - [Multiple sclerosis in childhood: our experience and a review of literature]. AB - INTRODUCTION: Multiple sclerosis (MS) is infrequent in childhood (0.3-2% of all cases of MS). At the present time more and more paediatric patients are being described. In this paper we describe our experience. MATERIAL AND METHODS: We review seven patients diagnosed as having MS before the age of 16, between 1984 and 1996. We have recorded: age, sex, personal and family history, form of onset and clinical course. CSF, neurophysiological and neuroimaging tests, treatment and diagnostic category according to the criteria of Poser. RESULTS: Four boys and three girls aged between three and thirteen years at the onset of the disorder. In one case there was a family history of MS. The most frequent form of onset was hemiparesia, followed by oculomotor paralysis and cerebellar disorders. Six cases followed a recurrent-remittent course and one followed a secondarily progressive course. There were oligoclonal bands (OGB) in the CSF in three cases. Visual evoked potentials (VEP) were abnormal in six cases. Magnetic resonance (RM) was a great help in diagnosis and in six cases was very informative. Six patients were treated with corticosteroids during the acute phase and two with long-term azathioprine. CONCLUSION: The commonest form of presentation, hemiparesia, makes the differential diagnosis with acute hemiplegia of childhood obligatory. Neurophysiological techniques, especially the VEP are very useful for initial assessment. RM is the most sensitive method, although it is not specific for diagnosis. The average follow-up period (4.5 years) is too short to determine the prognosis. PMID- 9736954 TI - [Neuroblastic migration: embryological aspects and mechanisms]. AB - INTRODUCTION: The migration of immature neurons of the cerebrum is genetically programmed from the primitive neuroepithelium before the end of the final mitotic cycle. The orientation of the mitotic spindle determines when a neuroepithelial cell is ready to start migration and the proportion of major genetic material it is destined to receive. DEVELOPMENT: The gene LIS1, defective in lissencephaly type 1 of Miller and Dieker, is expressed in the neuroepithelial cells, in the ependyma and the Cajal-Retzius neurons. These transitory fetal cells are the first neurons of the cerebral cortex. Most of the neurons of the cortical plate arrive by means of glial radial cells which guide them towards their destination. Cell adhesion molecules from the neuroblasts themselves, the glial radial cell, the extracellular matrix and perhaps the ependymal cells are important in adhering the neuroblasts to the glial radial cells. Genetic deficiency of these molecules results in defective migration. The mechanism of cellular movement is still not fully understood. Disorders of migration may also be induced by non genetic factors, such as infarcts or other lesions which damage or destroy the glial radial fibres during the fetal period. PMID- 9736955 TI - [The value of images in diagnosis of neuron migration disorders]. AB - OBJECTIVE: To present the fitest classification and the imaging peculiarities of the malformations of cortical development, most of which have been related with the epilepsy origin. METHODS: The study is based on an anatomical-histological classification scheme that shows three great groups of malformations of cortical development: 1. Malformations due to abnormal neuronal and glial proliferation. 2. Malformations due to abnormal neuronal migration. 3. Malformations due to abnormal cortical organization. RESULTS: The result of these abnormalities of the cortical development is the presence of several anatomical histological entities, actually perfectly identified by the magnetic resonance (MR), especially with the new high resolution methods. The most frequent entities, such as polymicrogyria, lissencephaly, pachygyria, schizencephaly, cerebral heterotopia (cortical, subcortical or subependymal), and other rarer types are reviewed according with the numerous references of the literature and the findings observed in the cases of our series of about one hundred patients which includes cases of every type of malformation. CONCLUSION: MR is a conclusive study in order to identify and classify the malformations of cortical development, most of which are associated with neurological disturbances: epilepsy, mental retardation, language and/or behavioral problems or motor dysfunction. PMID- 9736956 TI - [Clinical presentations, differential diagnosis and management of obstetric brachial palsy]. AB - INTRODUCTION: The brachial plexus originates from C5 to T1 spinal segments. The brachial plexus includes the ventral ramus, trunks, divisions, cords and branches. DEVELOPMENT AND CONCLUSIONS: Brachial plexus injuries produce clinical syndromes. The Duchenne-Erb syndrome is the most frequent presentation of obstetric brachial plexus injury. The differential diagnosis of brachial plexus palsy include decreased arm movements due to pain, or weakness caused by a lesion of the nervous system outside in the brachial plexus, or by a lesion in the brachial plexus due to non-obstetrical causes. Management of these patients initially includes considering the possibility of clavicular and humeral fractures and posterior subluxation of the shoulder; and subsequently considering the possibilities of subscapularis muscle contraction or posterior shoulder subluxation in patients that develop internal rotation contracture of the shoulder; or flexion, pronation or supination contracture in patients with forearm deformation. Treatment consist of physical therapy, administration of botulinum toxin, electrical stimulation, neurolysis, nervatization, removal of neuromas and nerve grafting, treatment of fractures and subluxation, release of muscle contracture and tendon transplantation. PMID- 9736957 TI - [Neurophysiological assessment of children with obstetric brachial plexus palsy]. AB - INTRODUCTION: The objectives of the neurophysiological evaluation of infants with brachial plexus palsy are to determine the time of occurrence of the lesion, to locate the lesion and to determine its course. METHODS AND CONCLUSIONS: These objectives are achieved by studying affected upper extremity muscles by needle electromiography (EMG) and affected nerves by motor and sensory conduction studies. EMG is performed in the first week of life in those patients with brachial plexus palsy of unknown etiology to determine the age of the lesion for medico-legal reasons. EMG is performed before surgery for tendon transfer in the selected muscles to assure that they are normal. EMG and motor and sensory conduction studies are performed at the age of 3 and 6 months in infants with less than 4 muscle weakness to determine candidates for surgical exploration. Motor and sensory nerve conduction studies are performed intraoperative to determine the functional status of the affected axons and the best surgical procedure (neurotization, neurolysis and/or neuroma resection and homologous nerve graft). PMID- 9736958 TI - [Surgical treatment of children with brachial plexus paralysis]. AB - OBJECTIVE AND METHODS: A variety of surgical procedures exist for early repair of the nerve injury in obstetrical brachial plexus palsy, including neuroma excision and nerve grafting, neurolysis and neurotization. Secondary deformities of the shoulder, forearm, and hand can similarly be reconstructed using soft tissue and skeletal procedures. This review describes our surgical approach to maximize the ultimate functional outcome in infants and children with obstetrical brachial plexus palsy. PMID- 9736959 TI - [Risk factors in learning difficulties]. AB - OBJECTIVE: To review the literature of the risk factors in learning difficulties (LD) and to assess these factors in children diagnosed as having LD in the Neuropaediatric Department of the Hospital Universitario Infantil La Fe in Valencia, in 1996 and 1997. MATERIAL AND METHODS: A retrospective study was done of children diagnosed in the Hospital Infantil Universitario La Fe in Valencia as having learning difficulties. The different factors related to LD were assessed. RESULTS: Between 1 January 1996 and 31 December 1997 a total of 32 children were diagnosed as having learning difficulties. There were 62.5% boys and 37.5% girls of an average age of 10 years and 10 months. The average intelligence quotient was 82.1. The commonest LD was difficulty with reading and writing. CONCLUSION: Learning difficulties form a large proportion of the routine pathology seen in the Neuropaediatric Department. It must be remembered that in most children with learning difficulties there is undiagnosed underlying neurological pathology which becomes noticeable on reaching an age at which schooling becomes more demanding. PMID- 9736960 TI - [Neurological assessment of learning disorders]. AB - INTRODUCTION: The neurological concept of learning is approached from a cybernetic point of view, taking into account that a child should recognize a fact, learn it semantically and decided whether it is worth storing; the dynamic aspect of memory is the true motor of the ability to learn and all this is modulated by the attention factor. DEVELOPMENT: The neurological evaluation of learning disorders is based on clinical examination which includes the so-called minor signs of the noetic functions, specifically language, the praxes, gnosias, perceptive-motor function, laterality and the lexical, graphic and calculation functions together with the modulating element, mentioned above, of the level of attention with or without hyperactivity. These semiological elements are grouped into three major categories of syndromes: motor syndrome, dyslexic-dysgraphic dyscalculation syndrome and the hyperkinetic syndrome or attention deficit with hyperactivity. We also note the differential diagnosis. We review the neurophysiological biological markers (EEG and brain mapping, cerebral evoked potentials, neurometry) and those based on neuroimaging techniques (cerebral CT, MR, SPECT and PET). CONCLUSIONS: The contribution of neurological assessment is considered as part of the functions of a multi-disciplinary team which should deal with the diagnosis and treatment of children with learning disorders. PMID- 9736961 TI - [Treatment of learning disorders]. AB - INTRODUCTION: The child neurologist, developmental pediatrician or child psychiatrist involved in the care of children with learning disabilities has a crucial role that extends beyond diagnosis. DEVELOPING: Despite the lack of sound scientific studies on treatment interventions for children with learning disabilities there are specific interventions that are helpful in the remediation of children with learning disabilities. CONCLUSIONS: The role of the health professional in the treatment of learning disabilities includes: 1. Making specific recommendations regarding early intervention strategies; 2. Putting into a neurological perspective the types of educational interventions that will maximize a child's potential; 3. Discussing the role of medications in the management of learning disorders, as well as using medications in appropriate situations, and 4. Coordinating the multimodal approach which is essential to the long term treatment of all children with learning disabilities. PMID- 9736962 TI - [Management of the mitochondrial encephalopathies]. AB - INTRODUCTION: In the management of mitochondrial diseases, it is most important to know what mitochondrias are, how they function and the pattern of inheritance present on the different conditions affecting them. DEVELOPMENT: An overview of mitochondrial function, inheritance, clinical and biochemical classification is done, reviewing the different fatty acid oxidation, pyruvic metabolism and respiratory chain defects. Classification by age of onset is also done. Approach to diagnosis, clinical aspects, laboratory findings and available treatment is discussed. Some comments about the use of a neurometabolic database are discussed, where more than 150 conditions are described in detail. CONCLUSIONS: By using the search capabilities of the software, a search by age of onset, symptoms, signs and laboratory findings help in reaching the most probable diagnosis in a given child, with its recommended current management. PMID- 9736963 TI - [Clinical features of the peroxisomal disorders]. AB - The peroxisomal disorders belong to a group of inborn errors of metabolism due to malformation or malfunction of these subcellular organelles. Their clinical features vary with age. However, the commonest form presents in the syndrome of Zellweger with severe hypotonia, craniofacial dysmorphism, stippled calcifications, renal cortical cysts and liver dysfunction. By means of complementation very long tests 17 different genotypes have been identified and in 15 of these there were neurological changes. Diagnosis requires recognition of the clinical features, raised levels of very long chain fatty acids, and radiological and neuroimaging studies. PMID- 9736964 TI - [The role of new antiepileptic drugs in childhood epilepsies]. AB - INTRODUCTION: All over the last years an important number of new drugs to treat epilepsy have become available. Initially they were applied as an add-on therapy to conventional agents but their indications in monotherapy are already becoming defined. OBJECTIVE: Giving an updated view of the actual situation of these antiepileptic drugs (AED). DEVELOPMENT: We may refer mainly to those whose clinical applications are more clearly defined by the moment, namely, lamotrigine, vigabatrine, felbamate, gabapentin and topiramate. First we may review their modes of action, in most cases better known than those of conventional agents. Then we may refer to their side-effects which are not completely known, Finally we may refer to their specific indications in some types of seizures and epileptic syndromes. CONCLUSIONS: Their efficacy may have to be evaluated further in a large number of comparative trials with conventional drugs. Their indications both in monotherapy and in polytherapy are to be fully defined and their toxicity profile is far from being completely known. Therefore, a conservative policy seems still justified, the more so as the cost of these agents is extremely high as compared to most conventional drugs. PMID- 9736965 TI - [Epilepsy centers: classification and basic requirements]. AB - Over the last two decades the surgical treatment of epilepsies has become a commonly used and effective method for the treatment of intractable epilepsy. However, because of its wide use in various types of centers throughout the world the methodology involved varies significantly throughout the centers. With this article we propose the bases from which a multidisciplinary team of numerous centers can eventually develop an effective and acceptable international protocol for the surgical treatment of the epilepsies and the classifications of the centers. PMID- 9736966 TI - [Mucolipidosis: clinical and genetic aspects]. AB - INTRODUCTION: The concept of the mucolipidoses developed as the result of encounters with patients whose clinical appearance suggested a mucopolysaccharidosis, i.e. coarse facies, short stature, skeletal dysplasia, joint contractures and lysosomal accumulations but did not have an excess of mucopolysaccharides in their urine. DEVELOPMENT: This clinical phenotype embraces the early onset forms of sialidosis (mucolipidosis I), as well as I-cell disease (mucolipidosis II) and pseudoHurler polydystrophy (mucolipidosis III). Two disorders that were later added to the mucolipidoses, the Cherry-red Spot Myoclonus Epilepsy Syndrome (sialidosis type I) and mucolipidosis IV differ in that the facial appearance is normal and they do no have skeletal dysmorphism. Enzyme deficiencies have been identified in mucolipidosis I, II and III. The basic molecular cause of mucolipidosis IV is not known but linkage analysis is underway to identify the chromosomal map position of the defective gene. Precise diagnosis of these autosomal recessively inherited diseases depends upon radiographic studies for skeletal dysplasias, analyses of urine for sialyloligosaccharides, morphologic studies of bone marrow and skin biopsy specimens and enzymatic determinations of sialidase and UDP-N-acetylglucosamine: lysosomal enzyme N-acetylglucosamine-1-phosphotransferase. CONCLUSIONS: Case descriptions from the author's experience are presented to illustrate the use of a decision tree employing these diagnostic modalities. The existence of naturally occurring animal models for sialidase deficiency (SM/J mouse) and the phosphotransferase deficiency (mucolipidosis III cat) provide opportunities to develop new therapeutic approaches. PMID- 9736968 TI - Master index to volumes 61-70. PMID- 9736967 TI - Teratology Society 38th annual meeting, The Neurobehavioral Teratology Society 22nd annual meeting, and the 11th International Conference of the Organization of Teratology Information. June 1998. San Diego, California, USA. Abstracts. PMID- 9736969 TI - [Process of high performance training--exemplified by gymnastics]. AB - A controlled, long-term training process, extensive medical treatment as well as a satisfying social and familiar environment are the key factors for a healthy development of young athletes in high level gymnastics. PMID- 9736970 TI - [Effect of high performance sports on puberty development of female and male gymnasts]. AB - High impact training may influence the pubertal development of athletes. The effects of high intensity training on pubertal development of female and male elite gymnasts are presented. 22 female and 18 male elite gymnasts were enlisted in this study. Prepubertal and pubertal stages were determined and hormonal levels regarding the hypophyseal, gonadal and adrenal axes were measured. The LH/FSH ratio necessary for the advancement of full pubertal maturation was assessed at 13.9 amd 13.0 years in female and male gymnasts, respectively. Female gymnasts demonstrated, on average, a delayed bone age of 1.7 years compared to their chronological age, whereas both bone and chronological age were identical in male gymnasts. There were no significant pubertal increase of estrogen levels in female gymnasts during their peripubertal development indicating a low amount of fat mass in female elite gymnasts. Intensive physical training of elite female gymnasts combined with inadequate nutritional intake markedly affect pubertal development. These peripubertal effects are not observable in male gymnasts due to different training regimes in male and female elite gymnast. Regular monitoring of female gymnast during their vulnerable growth phase is necessary to minimize life-long physiological and psychological side effects of high impact training. PMID- 9736971 TI - [An increased risk of spinal injury in high performance gymnasts during pubertal growth?]. AB - Elite womens gymnastics has generally been described to induce vertebral overload injuries, especially in the pubertal growth phase. This paper describes the results of a 4-year prospective longitudinal and transversal investigation of German elite womens gymnasts with respect to the spine. The main results are: 1) Pathological changes are detectable by clinical investigation before X-ray abnormalities can be found. 2) A sensitive test is the spine springing test. 3) Functional abnormalities can be detected especially in the lumbosacral region. 4) There were no spondylolyses. 5) Osteochondrotic lesions are frequent. They are concentrated in the dorsolumbar region. PMID- 9736972 TI - [Effect of high performance sports on energy metabolism]. AB - The knowledge about metabolism and muscular fatigue has been considerably improved during the recent years. Intramuscular pH should not generally be discussed as a factor of cellular fatigue as it has been shown to increase transiently at the beginning and to be very differently affected in ST-(6,9) and FT-fibers (6,2) at the end of exercise. During maximum exercise, we assume changes of muscle membrane potential due to increasing interstitial potassium concentrations as an important performance-limiting factor. The role, lactate is playing during exercise, has to be thought over. Its production by the binding of two protons to the pyruvate molecule is necessary for an intensive anaerobic and aerobic metabolism. The lactate molecules itself represent a source of energy, which is preferentially used by the heart muscle and by ST-fibers working at lower intensity. During long term endurance events, the glycogen stores are assumed to be a limiting factor. However, no direct casual relationship between glycogen and fatigue mechanisms has been found hithertoo. The anaerobic glycolytic metabolism decreases during long lasting exercise as a result of lowered glycogen stores. Therefore, metabolic acidosis cannot be observed after ultra-long endurance events. In contrast, a respiratory alcalosis is the common finding. PMID- 9736973 TI - [Nutritional behavior of female and male high performance gymnasts]. AB - Elite gymnasts pass through their whole physical and intellectual development with intensive physical training. In this period malnutrition can lead to delayed pubertal development with insufficient growth spurt and an increased incidence of stress fractures or osteoporoses. Different aspects about nutrition like body composition, objective and subjective eating-behaviour and sex-specific differences will be evaluated in our study. We examined 22 elite female gymnasts (age: median = 13.5 [12.0-16.1] years) und 19 elite male gymnasts (age: median = 12.3 [10.1-14.8] years). The following anthropometric measurements were carried out: weight, length, body mass index, upper arm circumference, arm muscle area, triceps skinfold, arm fat area. Eating diaries were compared with the recommendations of the German Federation of Nutrition and subjective eating behaviour was evaluated by questionnaires. Measurement of body composition showed an increase of musclemass at the cost of fatmass. The girls were smaller and leaner than the boys. Caloric intake in both groups was insufficient. Moreover the girls showed a tendency towards pathologic eating behaviour. PMID- 9736975 TI - [The German Orthopedic History and Research Museum]. PMID- 9736974 TI - [Anorexia nervosa and "anorexia athletica"]. AB - Available evidence supports the assumption that stressors in vulnerable adolescence potentially lead to restrictive dieting and imbalances of serotonergic metabolic particularly in females. In conjunction with idealized body images and developmentally characteristic bodily perceptions prone to distortion pathogenetic mechanisms of eating disorders are released. The entities of eating disorders are dimensionally viewed as points of continua a functions and categorized according to ICD-10 or DSM-IV, respectively. Data of our studies on grammar school students, ballet dancers and anorexia nervosa patients emphasize the necessity to differentiate different types of body shape by using the metric index. Facing this background a distinction of "anorexia athletica" appears unreasonable. PMID- 9736976 TI - [Ethipin refixation in osteochondrosis dissecans genu]. PMID- 9736977 TI - [Magnetic resonance tomography in congenital hip dysplasia and dislocation]. PMID- 9736978 TI - [Replantation of extracorporeally devitalized bone segments for defect reconstruction in tumor orthopedics--an overview]. AB - PURPOSE: The purpose of this paper is a review of the actual data about biological reconstruction by means of reimplantation of devitalised tumour bearing bone segments. REVIEW OF THE LITERATURE: Both experimental and clinical investigations proved extracorporeal autoclaving or high-dose irradiation to be a secure method for killing tumour cells. Animal experiments demonstrated that incorporation of devitalised bone segments took place after intramedullary nailing as well as after plate osteosynthesis. Bony union was followed by a slow revitalisation of the bone is reduced only after extremely high-dose irradiation. Autoclaving, however, may essentially impair biomechanics already after a short time of application high temperatures. The analysis of 115 patients with autoclaved re-implants and of 42 patients with irradiated reimplants showed encouraging functional long-term results when comparing to other biological reconstruction techniques. The time until consolidation was about 3 times longer than after osteotomies with well vascularised fragments on both sides. According to the data of the literature, the rate of local recurrences, fractures or deep infections was not higher than after other limb salvage procedures. CONCLUSION: Re-implantation of extracorporeally devitalised tumour bearing bone segments may be an excellent alternative reconstructive procedure in limb salvage for a small number of well selected patients. Stringent indication is most important for good results. Only patients who are willing and able for the differentiated aftertreatment should be considered and tumours with osteolyses essentially compromising stability should be excluded. Apart from general oncological necessities like adequate surgical margins, biomechanically correct osteosynthesis is most important in this alternative of reconstruction. PMID- 9736979 TI - [Magnetic resonance tomography in therapy follow-up after repositioning treatment of congenital hip dislocation]. AB - INTRODUCTION: After treatment of infantile hip dislocation in terms of reduction and retention in plaster casts or splints a problem of therapy controlling exists. To assess if the femoral head is in correct position centered in the socket ultrasound is not possible, because one cannot achieve the necessary standard position. Computertomograms are associated with a hereditary taint of radiation and plain roentgenograms lack of presentation of the important cartilaginous structures. METHOD: To control infantile hips after open or closed reduction from 1990 until 1996, 43 examinations by MR imaging were performed in 34 children. RESULTS: In our series MRI was able to provide exact information about the position of the femoral head independently of its state of ossification. Also, a clearly visualisation of the different structures of the acetabular fossa, particular acetabulum, pulvinar, bony and cartilaginous acetabular rim and limbus was possible. Furthermore the MR images showed interpositioned soft tissue, intraarticular effusion and displayed cartilaginous parts of the acetabular rim. CONCLUSIONS: MRI is an exact method to assess the anatomical and pathological conditions of the childlike hip joint. Because of the disadvantages of CT and X-ray, MRI is superior in terms of controlling the results after treatment of infantile hip dislocation. PMID- 9736980 TI - [Magnetic resonance tomography in diagnosis and therapy follow-up of patients with congenital hip dysplasia and hip dislocation]. AB - AIM: In patients with congenital dislocation of the hip the assessment of the correct position of the hip joint after closed or open reduction is very difficult to make from the radiograph with the hips in plaster. As the delayed recognition of a recurrent hip dislocation has bad effects on the outcome of the affected hip a safe and reliable imaging method must be employed. METHOD: From 1993 to 1996 6 patients with 8 congenital dislocations of the hip joint were examined by magnetic resonance imaging for evaluation of the position of the hip in plaster after reduction. Magnetic resonance imaging was performed immediately after closed or open reduction. 3 hips had to be treated by open surgery. RESULTS: The investigation confirmed that magnetic resonance imaging allows perfect differentiation between the bony and cartilaginous parts of the hip joint in plaster as well. Interpositioning of soft tissues which prevent reduction could also be visualized clearly. The best sequence in order to differentiate bony from cartilaginous structures was a gradient echo sequence in flash technique using a flip-angle of 60 degrees. In all cases the correct position of the hip joint after reduction could be demonstrated in plaster. CONCLUSION: Therefore, magnetic resonance imaging is the imaging method of choice for confirmation and documentation of the reduced position of the hip joint in plaster. Radiographs are no longer needed. PMID- 9736981 TI - [Deformities and misalignment of feet in children--a field study of 345 students]. AB - PURPOSE: purpose of this study was to evaluate paediatric feet and to document possible problems. MATERIAL AND METHODS: In 345 high school pupils of the 5th and 6th class aged 10 to 13 the feet as well as leg alignment were evaluated with a standardised protocol. RESULTS: Only 36.5% showed regular feet. In almost 2/3rd of the pupils malalignements or even deformities were present. The valgus foot showed the highest incidence with 39.4% followed by a flat foot with 19.1%. 17.1% of the children showed a hallux valgus. There was no correlation between body weight, body height, leg alignment and foot deformity. However, a significant correlation could we found between hallux valgus and splay foot. CONCLUSION: The presented data underlines the necessity for paying attention on the problems of the paediatric feet. During sport lessons in school a special foot training for children should be performed. PMID- 9736982 TI - [Prenatal ultrasound diagnosis of children with dysmelia]. AB - Since 1979 prenatal ultrasonography is a screening method in Germany. The examination of head, thorax and femur is now established as a routine method. AIM OF THE STUDY: The evaluation of the effectiveness and the consequences of an early prenatal diagnosis of foetal limb deficiencies. METHOD: From 1989-1994 we had 403 children with 207 minor or 196 major limb deficiencies. Questionnaires were sent to those 196 parents of children with major diseases 134 cases (68%) of children with limb deficiencies could be analysed. The average age of the mother at birth was 27.5 years and the age of the father was 30.5 years. RESULTS: 61 children had deficiencies of the upper limb. another 61 of the lower limbs and 12 had combined defects of the limbs. The mothers had 5.6 (1-15) prenatal ultrasounds examinations. In 63% (n = 82) these were performed by the obstetric practitioners, in 30% in departments of obstetrics and 7% in a university hospital. In 5.2% only, the limb deficiency was diagnosed antenataly. CONCLUSION: If there are any unclear findings the women should be sent to a centre of prenatal diagnostics. In cases of congenital deficiencies of the limbs there is the possibility] for the parents to combined to combined consultation of the obstetrician, the human genetic and the orthopaedic to plan the treatment and talk about all consequences. PMID- 9736983 TI - [Reference values for development of the iliac crest apophysis (Risser sign)]. AB - PURPOSE: Actualization of the standard values for the maturation of the iliac bone apophysis by Risser. METHODS: The study analysed retrospectively 515 data from 154 patients of german descent with idiopathic scoliosis. The study group included 312 X-rays of 82 female and 203 X-rays of 72 male patients, born between 1970-1985 and within the age of 10 and 20 years at the time of the evaluation. The middle chronological age, the standard deviation, the difference between female and male patients as well as the t-value were calculated for each stage. In addition the intervals with the corresponding t-values were calculated. RESULTS: The middle chronological age calculated for the stages of Risser shows significant differences. Because of acceleration the ossification begins earlier and finishes later. As assumed, the standard values were different for female and male patients. CONCLUSION: The necessity of a new and complete investigation was given because of the acceleration of the maturation and the incomplete existing standard values. CLINICAL RELEVANCE: This study contains current standard values for maturation of the iliac bone, which could be applied for analysing the skeletal maturation. PMID- 9736984 TI - [Proprioceptive abilities of patients with post-traumatic instability of the glenohumeral joint]. AB - PURPOSE: The purpose of this study was to evaluate joint position sense (JPS) in patients with posttraumatic glenohumeral instability. MATERIALS AND METHODS: In 28 patients with posttraumatic instability and in a matched control group of 30 subjects proprioception capability was evaluated. For documentation of proprioception an angle reproduction test (ART) was performed with which joint positions sense (JPS) was measured for abduction, flexion, and rotation in three angles each. RESULTS: In both groups there was a significant better JPS with visual control than without. In contrast to the control group the patients were not able to increase angle reproduction capability without visual control when comparing positions below shoulder level with positions at or above shoulder level. When comparing the patients to the controls there were differences in most of the ARTs with worse results in the patient group. These differences were significant in 150 degrees flexion with and without visual control, in 150 degrees abduction without and in 100 degrees abduction with visual control. For rotation there were trends for almost all joint positions, however, the differences were significant only in the -45 position. When comparing the noninjured contralateral shoulder of the patients with the control group, there still were differences. Again these were not in all joint positions significant, but significant worse JPS could be demonstrated in 150 degrees abduction without visual control, 50 degrees flexion without visual control, -45 degrees rotation without and 0 degrees rotation with visual control. CONCLUSIONS: A proprioceptive deficit can be documented in patients with posttraumatic glenohumeral instability. This may be one reason for permanent instability. The contralateral joint also shows reduction in joint position sense. For consecutive treatment as well as for rehabilitation both shoulder joint should be addressed. PMID- 9736985 TI - [Value of glenoid osteotomy in treatment of posterior shoulder instability]. AB - Purpose of this retrospective study was to evaluate the midterm results of glenoid osteotomy in the treatment of posterior instability and furthermore to evaluate the relevance of etiopathogenesis on the outcome. METHOD: Between Jan. 83 and May 94 32 patients underwent surgery due to posterior instability. Retrospectively these patients were classified based on their etiopathogenesis. 28 patients could be reexamined clinically and radiologically after and average of 4.85 years, of which 21 were treated with a glenoid osteotomy. For evaluation the Constant-Murley and the Rowe-Score were used. RESULTS: The Constant-Score showed 82,2% and the Rowe-Score 71,5% good or excellent results. Evaluation of glenoid osteotomy demonstrated for the patients with atraumatic genesis an excellent result in 76.9% with a recurrence rate of only 15.4%. On the other hand patients with traumatic instability showed a recurrence rate of 50%. The radiologic assessment demonstrated degenerative changes in nearly 30% of cases treated with glenoid osteotomy. CONCLUSION: This study shows that glenoid osteotomy can provide good result in the surgical treatment of posterior atraumatic instability. For treatment of traumatic instability, however, glenoid osteotomy is not the procedure due to high recurrence and arthrosis rates. RELEVANCE: Posterior glenoid osteotomy should be reserved for patients with atraumatic posterior instability combined with an increased retroversion angle of the glenoid especially because of the high rate of arthrosis. PMID- 9736986 TI - [Long-term results of basal wedge osteotomy in metatarsus primus varus in the young patient]. AB - INTRODUCTION: Aim of this retrospective study was to analyse the long term results after basal closing wedge osteotomy for correction of metatarsus primus varus and hallux valgus in the younger patient. PATIENT AND METHODS: 49 patients (70 feet) were operated according to a basal closing wedge osteotomy from 1974 to 1985 at our institution. Age was under 40 years in all patients at the time of surgery. 34 patients (50 feet) were evaluated in respect to their clinical and 26 patients (37 feet) to their radiological outcome. The average age was 26 years (14-39 years). The follow-up was 12 to 22 years (Median: 18 years). Analysis was performed using the patient's record, weight-bearing X-rays, a standardized questionnaire and clinical investigation. RESULTS: 82% of the patients had very good and good subjective results. Cosmetics was rated very good and good in 78%, 88% of the patients were painfree. Radiological analysis at follow-up: Hallux valgus-angle 19,3 degrees, intermetatarsal I/II-angle 6 degrees, shortening of first metatarsale 5 mm, at average; dorsal elevation of first metatarsale 38%, degenerative arthritis of the metatarsocuneiforme joint 19%, congruency of first metatarsophalangeal joint 54%,sesamoid subluxation: 46% grade 0, 30% grade I, 14% grade II and 10% grade III. In 14 feet (28%) metatarsalgia was found. DISCUSSION: The basal closing wedge osteotomy is rather a technically demanding procedure conjuncted with a higher risk of failure. Satisfactory long term results can be obtained by an ideal operating technique. As undesirable side effects shortening of the first ray and dorsal malangulation of the first metatarsale may occur consecutively leading to metatarsalgia. Lower risk procedures like the crescentic osteotomy according to Mann or chevron osteotomy should be preferred. PMID- 9736987 TI - [Long-term outcome of arthroplasty of the first metatarsophalangeal joint]. AB - INTRODUCTION: Resection arthroplasty of the first metatarsal-phalangeal joint is a wellknown operation for hallux valgus. The follow-up results more than 17 years after arthroplasty are recorded and discussed. MATERIAL AND METHOD: Between 1971 and 1980, 335 arthroplasties were performed on 205 patients. The only indication for arthroplasty of the great toe was hallux valgus. The technique of arthroplasty was according to Keller-Brandes. 102 patients were reexamined clinically and radiographically, after 17.6 years on average. RESULTS: Questionnaire assessment revealed a significantly prolonged walking distance and 70% of the patients were painfree at the time of investigation, whereas 67% suffered from severe pain before operation. Clinical evaluation showed diminished weightbearing of the great toe during walking and a reduced range of motion of the partial-resected first metatarsal-phalangeal joint. Radiographically the shortening of the phalangeal bone of 37% was evident as expected. Hallux-valgus angle was 23 degrees at the time of investigation and 34 degrees preoperatively in the mean. DISCUSSION: The high rate of hallux valgus relapse, especially due to a high intermetatarsal angle demonstrate unsatisfactory longterm results by the Keller-Brandes operation. We now recommend this operation for older patients and a differentiated approach according to the clinical and radiographical situation for younger patients. PMID- 9736988 TI - [Is skeletal alkaline phosphatase a valid staging marker in detection of osteoblastic skeletal metastases of prostate carcinoma?]. AB - PURPOSE: For patients with prostate cancer (CaP) the proof of osteoblastic bone metastases is decisive regarding the prognosis as well as the therapeutical concept. To evaluate the efficiency of skeletal alkaline phosphatase (SAP) as staging marker for bone metastases in prostate cancer, SAP was measured in CaP patients with and without bone metastases compared with prostate-specific antigen (PSA) as the marker of choice till now. METHOD: 73 patients with histological proven, but still untreated CaP were entered into the study. After staging the patients were divided into 3 groups: group I: patients with CaP and bone metastases (n = 21), group II: patients with locally advanced CaP without bone metastases (n = 26), group III: patients with clinically localized CaP without bone metastases (n = 26). Serum concentration for SAP and PSA were determined using radioimmunassay. As reference range we defined serum concentrations for SAP < 19 ng/ml and for PSA < 100 ng/ml. RESULTS: 71% of the patients with bone metastases (group I) showed elevated SAP and PSA serum concentrations. In contrast, patients without bone metastases (group II + III) have normal SAP values (<19 ng/) and in 19% of the cases elevated PSA-values (>100 ng/ml). This resulted in a sensitivity and specificity of 71% and 100% for SAP and 71% and 81% for PSA. The positive predictive value for osteoblastic bone metastases was 100% for SAP and 60% for PSA. CONCLUSION: SAP is a useful staging marker in prostate cancer and can contribute for an early detection of osteoblastic bone metastases. PMID- 9736989 TI - [Bone density--reference values in German men. A study of the lumbar spine with the Lunar-DPX-densitometer]. AB - The DXA-technique is a well established method to study bone mineral density (BMD). Until now there are no reliable reference data based on the male population of Germany. QUESTION: Are the data base, given by the manufacturer transferable to the male population in Germany? METHODS: So a cross sectional study based on 715 healthy males with German ethnic background (age 20-89) was carried out. Comparison was made to the ap spine reference data of northern Europe. RESULTS: The peak bone mass was 1,26 +/- 0,17 g/cm2 at the age of 20-24 years. After that the BMD decreases to the sixth decade (1,09 +/- 0,18 g/cm2), further on there is an increase to 1,18 +/- 0,21 g/cm2 in the ninth decade. CONCLUSION: Looking at a comparable Swedish study there are no significant differences, looking at a Finnish study there are statistically significant differences in the sixth decade (t = 3,246) and seventh decade (t = 2,413). The results of our study complete the data base until now, but one should be careful to transfer the data base provided by the manufacturer. PMID- 9736990 TI - [Development of gentamicin-resistant Small Colony Variants of S. aureus after implantation of gentamicin chains in osteomyelitis as a possible cause of recurrence]. AB - AIM: Recently, S. aureus small colony variants (SCVs) were reported to persist within cultured endothelial cells and to cause persistent and antibiotic resistant infections in humans. Because gentamicin can very reproducably select for electron transport deficient SCVs as shown in earlier in vitro experiments, we searched for SCVs in a patient with chronic osteomyelitis, who received gentamicin beads. METHOD: Special culture and identification procedures for determination of SCVs were used, including testing of the S. aureus specific nuc gene, pulsed field gel electrophoresis (PFGE) as a typing method and characterization of the auxotrophism of the SCVs. RESULTS: In a case of a 34-year old patient with chronic osteomyelitis who had previously been treated with gentamicin beads, menadione auxotrophic S. aureus SCVs as well as wild type S. aureus were recovered in multiple bone specimen. All different colony types isolated from simultaneous or from sequential specimen were shown to be clonal by PFGE. CONCLUSION: Recovery of S. aureus SCVs from a patient treated with gentamicin beads suggests that the slow release of gentamicin into the local environment may be an efficient way to select for and/or induce SCVs. These data should alert physicians to also consider SCVs when a treatment failure occurs in a patient that has received gentamicin beads. PMID- 9736991 TI - [Analysis of orthopedic specialty knowledge and comparison of 2 written test methods]. AB - QUESTION: What about the knowledge of specific orthopaedic subjects in medical education as tested with mc-questions and open-ended questions? METHOD: 524 medical students were evaluated by a questionnaire survey to determine the state of orthopaedic knowledge in different periods of medical education. With the same questionnaire the use of two different types of written tests of knowledge (multiple choice questions and open-ended questions) has been tested and the results have been compared. The students participating in the orthopaedical practical (6th clinical semester) have been tested before and after the practical, given as pretest and posttest. RESULTS: The results show that the orthopaedic knowledge corresponds to the educational level so that during medical formation, there is a gradual knowledge gain. Comparison of the results of the pretest and the posttest reveals that the highest knowledge gain occurred during the orthopaedic practical. For all groups candidates performed better in mc questions than in open-ended questions. Likewise in all groups, open-ended questions were found to be more difficult than the mc-questions. CONCLUSIONS: To summarize it may be concluded that there is a gradual orthopaedical knowledge gain during medical formation. It was found that students gain a considerable part of their technical knowledge during the practicals. Both mc-questions and open-ended questions are useful to measure this knowledge gain where there are hints that open-ended questions give a more accurate assessment of the actual knowledge. The pre- and posttest setting can provide both teachers and students with feedback regarding the realization and acquisition of instructional contents. PMID- 9736992 TI - [Lipoatrophia semicircularis--an orthopedic diagnosis?]. AB - INTRODUCTION: Lipoatrophia semicircularis (LAS) represents an isolated fatty atrophy usually located at the lower extremity. The atrophy is circular and painless and usually resolves without therapy. The skin modifications occur within a few days or weeks. The etiology is unknown. CASE: In the following we present a case of LAS with a segmental dysfunction of the third ipsilateral lumbal root of a 43-year-old women. It was possible to provoke a referred pain into the area of atrophy by passive motion of the lumbal spine into the plane of resistant. CONCLUSION: LAS is rare and slightly known among orthopedists. Knowledge of the typical morphological modifications will lead to diagnosis. In most of the cases further diagnostic measures are not necessary. We recommend a segmental investigation of the spine in the case of secured diagnosis. PMID- 9736993 TI - [Comment on Weber, M., M. Morgenthaler: The significance of expansion of the occupational disease regulation for evaluating intervertebral disk damage]. PMID- 9736995 TI - Pyrrolizidine alkaloids. PMID- 9736996 TI - Bufadienolides of plant and animal origin. PMID- 9736997 TI - Fatty acid signaling in Arabidopsis. AB - Many organisms use fatty acid derivatives as biological regulators. In plants, for example, fatty acid-derived signals have established roles in the regulation of developmental and defense gene expression. Growing numbers of these compounds, mostly derived from fatty acid hydroperoxides, are being characterized. The model plant Arabidopsis thaliana is serving a vital role in the discovery of fatty acid derived signal molecules and the genetic analysis of their synthesis and action. The Arabidopsis genome sequencing project, the availability of large numbers of mutants in fatty acid biosynthesis and signal transduction, as well as excellent pathosystems, make this plant a tremendously useful model for research in fatty acid signaling. This review summarizes recent progress in understanding fatty acid signaling in A. thaliana and highlights areas of research where progress is rapid. Particular attention is paid to the growing literature on the jasmonate family of regulators and their role in defense against insects and microbial pathogens. PMID- 9736998 TI - The CURLY LEAF gene controls both division and elongation of cells during the expansion of the leaf blade in Arabidopsis thaliana. AB - The CURLY LEAF (CLF) gene in Arabidopsis thaliana (L). Heynh. is required for stable repression of a floral homeotic gene, AGAMOUS in leaves and stems To clarify the function of CLF in organ development, we characterized clf mutants using an anatomical and genetic approach. The clf mutants had normal roots, hypocotyls, and cotyledons, but the foliage leaves and the stems had reduced dimensions. A decrease both in the extent of cell elongation and in the number of cells was evident in the clf mutant leaves, suggesting that the CLF gene might be involved in the division and elongation of cells during leaf morphogenesis. An analysis of the development of clf mutant leaves revealed that the period during which tell division or cell elongation occurred was of normal duration, while the rates of both cell production and cell elongation were lower than in the wild type. Two phases in the elongation of cells were also recognized from this analysis. From analysis of an angustifolia clf double mutant, we found that the two phases of elongation of leaf cells were regulated independently by each gene. Thus, the CLF gene appears to affect cell division at an earlier stage and cell elongation throughout the development of leaf primordia. PMID- 9736999 TI - Characterization of rbcS genes in the fern Pteris vittata and their photoregulation. AB - The fern Pteris vittata L. belongs to the evolutionarily highest group of vascular plants that still maintains a free-living gametophytic stage. The two dimensional gametophytes developed under blue light exhibit higher CO2 fixation efficiency and different ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) small subunit (SSU) composition when compared to the red-induced filamentous gametophytes (H. Eilenberg et al., 1991, Plant Physiol 95: 298-304). To unravel the correlation between SSU structural differences and light regulation, two rbcS genes and two additional partial cDNAs were characterized. Fern rbcS genes resemble those of higher plants in their promoter light regulatory elements (LREs) and intron number and positions. However, the primary structure of the fern mature SSUs displays much higher divergency within the gene family. This structural variability was correlated with differential steady-state mRNA levels under red and blue light. Genes rbcS-1 and-4 and 4-to 6-fold higher transcript levels in red light while rbcS-2 and-3 contribute relatively more to the blue rbcS mRNA levels. Five of the 12 amino acids that differ between rbcS-2 and-4 affect hydrophobicity and might play a crucial role in determining the efficiency of CO2 fixation. Dendrograms of Rubisco SSUs and LSUs indicate early divergence of the fern types from the rest of the vascular plants. However, prominent higher-plant-like Rubisco features such as high carboxylation efficiency, promoter LREs and exon-intron structure, suggest that molecular specialization of the higher-plant Rubisco prototype occurred earlier than the emergence of ferns. PMID- 9737000 TI - Cell cycle regulation by plant growth regulators: involvement of auxin and cytokinin in the re-entry of Petunia protoplasts into the cell cycle. AB - In order to understand the mode of action of auxins and cytokinins in the induction of cell division, the effects of the two plant growth regulators 2,4 dichlorophenoxyacetic acid (2,4-D) and N6-benzyladenine (BA) were investigated using mesophyll protoplasts of Petunia hybrida, cultivated in either complete medium or in medium deficient in cytokinin, auxin or both. Firstly we studied DNA synthesis, using 5-bromodeoxyuridine/bisbenzimide Hoechst/propidium iodide flow cytometry analyses and by the monitoring of histone H4 transcript levels. Roscovitine, a cyclin-dependent kinase (CDK) inhibitor, was found to block the cell cycle prior to entry into the S and M phases in the cultured P. hybrida protoplasts. This suggests that in Petunia cell there is a requirement for CDK activity in order to complete the G1 and G2 phases. Further experiments using roscovitine showed that neither 2,4-D nor BA were individually able to induce cell cycle progression beyond the roscovitine G1 arrest. We also monitored the phytohormonal induction of S phase by studying variations in transcript levels of the gene for mitogenactivated protein kinase (PMEK1) and transcript levels of the cell division cycle gene cdc2Pet. Only 2,4-D, and not BA, was able to stimulate PMEK1 gene transcription; thus, the more rapid S-phase induction in 2,4-D-treated protoplasts may be attributable to the activation of this transduction pathway. In contrast, both plant growth regulators were required to induce the appearance of cdc2Pet mRNA transcripts prior to S-phase engagement. PMID- 9737001 TI - Functional analysis and cell-specific expression of a phosphate transporter from tomato. AB - For a better understanding of the molecular and biochemical processes involved in orthophosphate (Pi) uptake at the root/soil interface, we cloned a Pi-transporter c DNA (LePT1) from a root air-specific cDNA library of tomato (Lycopersicon esculentum Mill.). The corresponding protein belongs to the growing family of ion transporters with twelve putative transmembrane domains. It is highly homologous to recently isolated Pi transporters from higher plants, yeast and fungi. When expressed in a Pi-uptake-deficient yeast mutant, the L. esculentum phosphate transporter 1 (LePT1) protein exhibits an apparent Km of 31 MicroM. The transporter is still active at submicromolar Pi concentrations and mediates highest Pi uptake at pH 5. The activity of LePT1 is dependent on the electrochemical membrane potential mediated by the yeast P-type H + - ATPase. Transcript levels of LePT1 in tomato seedlings are detectable in all vegetative organs under Pi-sufficient conditions, with highest concentrations in root hairs. In situ hybridization studies demonstrate cell-specific expression of LePT1 in the tomato root. The LePT1 mRNA is detectable in peripheral cell layers such as rhizodermal and root cap cells. Under Pi-deprivation condition, mRNA levels are also detectable in young stelar tissue. This work presents molecular and biochemical evidence for distinct root cells playing an important role in Pi acquisition at the root/soil interface. PMID- 9737004 TI - Clustering of the nitrite reductase gene and a light-regulated gene with nitrate assimilation loci in Chlamydomonas reinhardtii. AB - Two new loci have been found to be clustered with five other genes for the nitrate assimilation pathway in the Chlamydomonas reinhardtii genome. One gene, located close to the 3'-end of the high-affinity nitrate transporter (HANT) gene Nrt 2; 2, corresponds to the nitrite reductase (NiR) structural gene Nii1. This is supported by a number of experimental findings: (i) NiR-deficient mutants have lost Nii1 gene expression; (ii) Nii1 mRNA accumulation is co-regulated with the expression of other structural genes of the nitrate assimilation pathway; (iii) nitrite (nitrate) utilization ability is recovered in the NiR mutants by functional complementation with a wild-type Nii1 gene; (iv) the elucidated NII1 amino acid sequence is highly similar to that of the cyanobacterial and higher plant enzyme, and contains the predicted domains for plastidic ferredoxin-NiRs. Thus, the mutant phenotype and the mRNA sequence and expression of the Nii1 gene have been unequivocally related. Accumulation of mRNA for the second locus identified. Lde1 (light-dependent expression), was not regulated by nitrogen, but like nitrate-assimilation clustered genes, its expression was down-regulated in the dark. PMID- 9737005 TI - Evolution and animal welfare. AB - Animal welfare is a topic often thought to reside outside mainstream biology. The complexity of the methods used to assess welfare (such as health, physiology, immunological state, and behavior) require an understanding of a wide range of biological phenomena. Furthermore, the "welfare" of an animal provides a framework in which a diversity of its responses can be understood as fitness enhancing mechanisms. Different methods for assessing animal welfare are discussed, with particular emphasis on the role of an animal's own choices and reinforcement mechanisms. No part of biology is as yet able to explain consciousness, but by confronting the possibility that nonhuman animals have conscious experienced of suffering, animal welfare studies force a consideration of even this hardest problem of all biological phenomena in a particularly direct and evolutionary way. PMID- 9737006 TI - The effect of organic solvents on the determination of cyclic boronates of some beta-blockers by gas chromatography/mass spectrometry. AB - Formation cyclic boronates for beta-blockers, by use of triethylamine (TEA) and pyridine as catalysts, gives more effective product yield. Eleven beta-blockers, formed by cyclic boronation with n-butylboronic acid and TEA, produced higher yields than by on-column thermal reaction and seven beta-blockers were shown to give the highest yields in the phenyl cyclic boronation with pyridine or TEA by on-column thermal and general reaction. The phenyl cyclic boronate of nadolol produced one peak in the chromatogram and the n-butyl cyclic boronate showed two peaks. On-column derivatization and n-butylboronic acid and pyridine was effective in saving analysis time and for convenience, even though some n-butyl cyclic boronates by cyclic boronation with TEA gave a better yield. In n-butyl cyclic boronation with pyridine by on-column thermal reaction the detection limits were 0.1 to 4 ng/microL in urine with a signal-to-noise ratio of 10:1. PMID- 9737007 TI - Identification of pollutants in a municipal well using high resolution mass spectrometry. AB - An elevated incidence of childhood cancer was observed near a contaminated site. Trace amounts of several isomeric compounds were detected by gas chromatography/mass spectrometry (GC/MS) in a concentrated extract of municipal well water. No matching library mass spectra were found and Fourier transform IR and NMR analyses were not feasible due to the low concentration of the compounds. Mass peak profiling from selected-ion-recording data (MPPSIRD) provided the sensitivity and scan speed necessary to acquire mass peak profiles at mass resolutions of 10,000 to 20,000 for the molecular ion (M+) and 10 fragment ions as capillary GC peaks eluted. Using a profile generation model (PGM), the elemental composition of the molecular ion was determined from the exact masses and abundances of the M, M + 1 and M + 2 profiles. Fragment ion compositions were determined from their exact masses based on the elements in the molecular ion. Exact mass differences between the molecular and fragment ions corresponded to unique combinations of atoms for the neutral losses. Consequent reduction of the number of possible structures for the fragment ions simplified mass spectral interpretation. After inspecting library mass spectra for smaller molecules, isomeric structures were hypothesized with cyano and alkylcyano groups attached to tetralin. A literature search found such isomers produced by an industrial polymer synthesis. Three isomers in a standard form polymerization of styrene and acrylonitrile provided the same mass spectra and GC retention times as isomers in the extract. PMID- 9737008 TI - Influences of pH and in-source collisional energy on the cationization of insulin. AB - The influence of pH and in-source collisional energy variations on interactions between a model protein and sodium or cesium ions has been studied by electrospray ionization mass spectrometry. Behavioural differences regarding cationization have been observed and seem to be linked to the neutral of deprotonated form of carboxylic acids. At pH 8, cesium and sodium adducts totally disappear in the case of highest charge states. Increasing the cone voltage leads to an overall change loss that can be attributed to the loss of cesium, sodium ions and even protons. PMID- 9737009 TI - Gas chromatography/mass spectrometry analyses of [2,3,3-d3] serine, [2,3,3-d3] cysteine and [3-13 C] cysteine in plasma and skin protein: measurement of transsulphuration in young sheep. AB - A new procedure using stable isotope labelled serine (L-[2,3,3-d3] serine) and cysteine (L[13-3-13 C] cysteine) and analysis by gas chromatography/mass spectrometry (GC/MS) has been developed to measure transsulphuration in sheep. The enrichments of the tracers in plasma and skin biopsy samples were measured by GC/electron impact MS analysis of the t-butyldimethylsilyl derivatives. The measured recoveries of the standards enriched with [3-13 C] cysteine from 0.1% to 8%, or with [2,3,3-d3] serine from 0.14% to 14% were greater than 99% of the theoretical values, and the variation coefficients were less than 3% when the enrichment was higher than 0.5%. The use of dithiothreitold (DTT) as a reducing agent before deproteinization of the sample and during the derivatizations successfully increased the cysteine peak area and significantly improved reproducibility in the analysis. The cysteine residues in protein from the skin biopsy were also during the protein hydrolysis with DTT in 6 n HCI. The method was applied to measure transsulphuration of methionine in young sheep. The amount of cysteine derived from transsulphuration accounted for 17% to 21% of the irreversible loss rate of cysteine, depending on the substrate supplies. The results are consistent with other reports. Compared with conventional methods of measuring transsulphuration using radioactive isotopes, the processes of animal experimentation was sample analysis were simple, and there were no radiation hazards. The method should prove useful in studies on the metabolism of methionine and cysteine in human and animals. PMID- 9737010 TI - In vivo pharmacokinetic screening in cassette dosing experiments; the use of on line Pprospekt liquid chromatography/atmospheric pressure chemical ionization tandem mass spectrometry technology in drug discovery. AB - Drug discovery is a fast growing field and the number of compounds generated daily by the pharmecutical industry is enormous. The necessity of developing new experimental strategies and analytical methods to rapidly screen the pharmacokinetics (PK) behavior of these compounds becomes a real challenge. A novel strategy to support in vivo PK screening in cassette doing experiments, using a fully automated system capable of analyzing between 320 to 960 samples a day by instrument in n-in-one experiment ( n = 64 in this work), has been developed. Using an on-line extraction technique, the average observed recovery was 64% using a single C18 procedure. A weighted (1/x) linear equation was used to perform standard calibration (0.5 to 500 ng/microL) and the average R value obtained was 0.994 (R2 = 0.997) for 63 analytes. The limit of detection, defined as a signal-to-noise ratio of 3 or greater, was found to be 25 pg for 41 of the 63 analytes (65%) and 250 pg for 57 of the 63 analytes (90%). The complete automation procedure using the Prospekt-LC-APCI/MS/MS system has substantially improved throughput in the area of drug discovery and bioanalysis. PMID- 9737011 TI - Genotyping of apolipoprotein E by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. AB - The genotyping of the various isoforms of Apolipoprotein E (apo E) has been performed using matrix-assisted laser desorption/ionization (MALDI-MS). The polymerase chain reaction was used to amplify the specific apo E gene sequence followed by digestion with Cfo I (Clostridium formicoaceticum), for generating restriction fragments for rapid and accurate mass analysis. An exonuclease I digestion step was introduced to remove the unused primers after PCR, which can otherwise interfere in the mass spectral analysis. By replacing the gel electrophoresis detection step with MALDI-MS, restriction isotyping of the apo E gene was achieved. Genotyping of an unknown sample and obtained from an independent diagnostic laboratory demonstrated the validity of the MALDI-MS method for the routine analysis of apo E. PMID- 9737012 TI - Liquid-liquid extraction procedure for trace determination of cyclophosphamide in human urine by high-performance liquid chromatography tandem mass spectrometry. AB - A sensitive, specific and accurate high performance liquid chromatography/ionspray-tandem mass spectrometry procedure (HPLC/MS/MS) has been developed to quantify cyclophosphamide in human urine from hospital personnel involved in drug preparation and administration of antineoplastic alkylating agents. This methodology, which includes liquid-liquid extraction with ethylacetate, requires no derivatization procedures, preventing cyclophosphamide (CP) from possible thermal and chemical decomposition reactions. We detected the excretion of this unmetabolized alkylating drug in 50% of all the study participants. The amount of CP ranged from 0.1 ng microL-1 to 1.9 ng microL-1 urine. This methodology was validated by the use of ifosfamide as internal standard. The assay was linear over the range 0 to 3.2 ng microL-1 urine, with a lower limit of quantification of 0.2 microL-1. The limit of detection was assessed at 0.05 ng microL-1 urine. This method is characterized by a coefficient of variation < 10%. Standard calibration curves, obtained on three different days, had correlation coefficients always greater than 0.998. The intra and interday precision were within 11%, and accuracy was in the range 99-103%. The mean extracted recovery assessed at three different concentrations (0.5, 0.8, 3.2 ng microL-1) was always more than 85%. The extraction efficiency of cyclophosphamide from urine samples was also studied at six different pH values (pH 4, 5, 6, 7, 8, 10). The maximum extraction efficiency was obtained when the pH of urine solutions was adjusted to 7.0 PMID- 9737013 TI - Influence of stereoisomeric glucose, galactose and mannose residues on fragmentation at their glycosidic linkages in post-source decay fragment analyses for oligosaccharides using matrix-assisted laser desorption/ionization time-of flight mass spectrometry. AB - The isomeric sugar branched beta-cyclodextrin (CD) derivatives (Glc-beta CD, Gal beta CD, and Man-beta CD) were analyzed by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. In the MALDI post-source decay (PSD) fragmentation of Glc-beta CD, Gal-beta CD and Man-beta CD, the fragment ions of [M-Glc]+, [M-Gal]+ and [M-Man]+ were produced by the one site cleavage of the alpha 1-6 glycosidic linkage at the branch (Y-type fragmentation). The abundances of [M-Gal]+ ions were much higher than that of [M Man]+. These results indicated that Glc-beta CD and Gal-beta CD were distinguishable from Man-beta CD and that Y-type fragmentations of the branched glycosidic linkage at the glycosyl donors of alpha-D-Glc and alpha-D-Gal were more predominant than that at the glycosyl donor of alpha-D-Man by MALDI-PSD fragment analysis. It was concluded that the abundances of PSD fragment ions depended on the stereochemistry of the glycosyl donors in the cleavage of the glycosidic linkage of the oligosaccharides in MALDI-TOF mass spectra. PMID- 9737014 TI - Separation of cyclosporins by high-performance liquid chromatography and mass spectrometric study of cyclosporin metabolites. AB - A semi-preparative (high-performance liquid chromatography) method for separation of cyclosporin metabolites in a fungal fermentation sample was developed. By using the optimized chromatographic separation, 20 cyclosporin metabolites in a process sample were isolated, and their molecular masses measured by double focusing sector mass spectrometry. The structures of some of the cyclosporin congeners were investigated by tandem sector mass spectrometry using fast atom bombardment ionization. The isobaric cyclosporins D ([Val2] CS) and G ([Nva2] CS) were differentiated by significantly different relative intensities of a fragment ion at m/z 30 [CH4N]+ from a precursor ion at m/z 72 [C4H10N]+. Otherwise the tandem mass spectrometry (MS/MS) spectra of the fragment ions of the two isomers are very similar. Cyclosporin A and iso-cyclosporin A were also analysed by high energy MS/MS. No significant fragment ions were found to provide distinctions between them. Relatively good overviews of process samples containing very similar structures were achieved by combining LC separation, molecular ion recognition and MS/MS characterization. PMID- 9737015 TI - Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis of proteins in human cerebrospinal fluid. AB - Matrix-assisted laser desorption/ionization time-of-flight mass spectra of proteins in cerebrospinal fluid analyzed without prior purification are presented. Less than 100 fmol amounts of proteins in the 10,000 to 20,000 u mass range and linked to human disease (multiple sclerosis, Alzheimer's disease, and stroke) were detected in a complex mixture of proteins and peptides, in the presence of high concentrations of salts, lipids and free amino acids. The mass resolution was sufficient to distinguish between the non-hydroxylated and hydroxylated forms of a 13,400 u protein. Simple fractionation of the cerebrospinal fluid using microbore-reversed phase high performance liquid chromatography improved signal-to-noise ratios in the mass spectra. High-accuracy peptide mass mapping and database searching were utilized to confirm the identity of several proteins. The presented results show that matrix-assisted laser desorption/ionization time-of-flight mass spectrometry could be used as a tool to perform rapid screening of chemically altered proteins in small volumes of biological fluids. PMID- 9737016 TI - Isotope dilution-liquid chromatography/electrospray ionization-tandem mass spectrometry for the determination of serum thyroxine as a potential reference method. AB - A new candidate reference method is presented for the determination of thyroxine in serum. The method is based on isotope dilution-liquid chromatography/tandem mass spectrometry using electrospray for ionization. The internal standard used was 13C6-thyroxine, sample pretreatment consisted of protein precipitation and a two-step liquid/liquid extraction procedure, HPLC was performed on a C-18 column with an eluent containing methanol/water/formic acid (60:40:0.1, by volume), and finally thyroxine and its isotopically labeled analogue were measured in the selected reaction monitoring mode for the transitions m/z 777.7--> 731.7 and m/z 783.7--> 737.7, respectively. The detection limit for thyroxine was 6 pg, the within-run coefficient of variation was 1.1%. The samples were measured in six fold: in duplicate on three independent days. The mean overall coefficient of variation of the method was 1.6%. The new method was evaluated by measuring nine control sera previously determined by an existing ID-GC/MS method. The differences between the results of the two methods ranged from-1.6% to +3.3%, with a mean of +0.2%. PMID- 9737017 TI - Analysis of variant forms of porcine surfactant polypeptide-C by nano electrospray mass spectrometry. AB - Electrospray (ES) mass spectrometry has been used to analyse preparations of porcine pulmonary surfactant polypeptide-C (SP-C). A number of variant forms of the native 35-residue dipalmitoylated peptide were detected including (a) C terminally methylated SP-C, (b) C-terminally methylated and methionine oxidized SP-C, (c) N-terminally truncated, C-terminally methylated and methionine oxidized SP-c, (d) C-terminally elongated, C-terminally methylated and methionine oxidized SP-C, and (e) tripalmitoylated, C-terminally methylated and methionine oxidized SP-C. C-terminal methylation and methionine oxidation are probably a consequence of the sample handling procedure. The occurrence of the C-terminally elongated form of SP-C has implications for the in vivo processing of proSP-C and the Tandem mass spectrometry (MS/MS) was used to confirm the amino acid sequence of SP-C and the presence of palmitoyl groups covalently linked to the peptide. Some of the structures of the variant forms of SP-C were determined by MS/MS. PMID- 9737018 TI - Mechanical properties of L929 cells measured by atomic force microscopy: effects of anticytoskeletal drugs and membrane crosslinking. AB - To shed light on the architecture of the cytoskeleton, we used the atomic force microscope (AFM) to measure the elasticity, viscoelasticity, and plasticity of L929 cells. The initial elastic response (Young's modulus approximately 4,000 Pa) of the cells to an applied force was followed by a slow compression of the cytoskeleton (tau 1/2 approximately equal to 10 s). When force application was terminated, the cytoskeleton underwent a sudden partial decompression and a subsequent slow, incomplete recovery. The role of the cytoskeletal elements in cell mechanics was accessed in AFM measurements carried out on cells treated with cytochalasin D, nocodazole, or colcemid. Cytochalasin D treatment reduced both elasticity (approximately 45%) and cytoplasmic viscosity (approximately 65%), whereas cells treated with nocodazole or colcemid exhibited a marked increase in elasticity (approximately 100%) and a slight increase in viscosity (approximately 15%). The AFM force measurements also provided evidence that the cell membrane and the cytoskeleton are mechanically coupled. Tightly adherent cells were stiffer than cells that were loosely attached. Moreover, cells crosslinked with either glutaraldehyde, 3, 3'-dithiobis [sulfosuccinimidylpropionate] (DTSSP), or Concanavalin A were more rigid than untreated cells. It is of interest that cells crosslinked with Concanavalin A, but not DTSSP, displayed plastic behaviors that may reflect the induction of cytoskeletal reorganization by Concanavalin A. PMID- 9737019 TI - [Experimental study of metoprolol-induced side effects]. AB - To study side effects of drugs in preclinical as well as in postmarketing surveillance phase is very important. In our experiments the influence of metoprolol on carbohydrate- and lipid metabolism was investigated in male. Wistar rats. Metoprolol is a liposoluble beta1-selective adrenoceptor antagonists. We have calculated therapeutic dose which reduced heart frequency/min of the animals by 25%. This was 10 mg/kg. The blood glucose and triglyceride values of healthy rats are in the normal human domain. Blood glucose was high after the first metoprolol dose and increased further with continued treatment. Drug administration period comprised 16 days. At finishing experiments diminished glycogen content was measured which may be related to higher glucose output. In blood samples obtained one hour after last 16. metoprolol dose administration triglyceride values were high and HDL-C decreased. These data pertain to the development of a secondary hypertriglyceridaemia. Hyperglycemic and hypertriglyceridaemic responses were established with therapeutic doses regimen so they may be considered as unwanted effects. PMID- 9737020 TI - [Effect of physicochemical properties of polyethylene glycol derivatives on drug release from wax matrices]. AB - The purpose of the present study was to investigate the effect of the various types and amounts of polyethylene glycols on the potassium chloride release from wax matrices. Potassium chloride as a highly water-soluble model drug was embedded into wax containing surfactants to produce sustained release dosage form. Various kinds of polyethylene glycols were chosen to control the dissolution profile. The dissolution profile of the matrix samples was characterized by the Weibull distribution. The physico-chemical characteristics- surface tension, HLB value, refractive index--of the aqueous solutions of the polyethylene glycols were also determined. On the basis of the examined physicochemical characteristics of various polyethylene glycols it is possible to select the optimal concentration of surfactant to formulate matrix dosage forms of required dissolution profile. PMID- 9737021 TI - [Investigation of polysaccharides of Echinops species. Medicinal plant polysaccharides I]. AB - Polysaccharides isolated from Echinops sphaerocephalus, Echinops ruthenicus, Echinops exaltatus, Echinops commutatus and Echinops orientalis (Asteraceae family) were investigated to study their qualitative and quantitative relations. The extracts of these studied plants have been used in traditional Chinese medicine as drugs with anti-inflammatory effect and anti-tumor promoting action in the osseous system [1]. We suppose that these biological effects are related to the polysaccharides especially to the acidic ones. The plant material was successively extracted with boiling water and six fractions were collected. The yield was 40-45%. The uronic acid content of these fractions, measured by the Bitter method, was relatively high 15-35% (Table I.). In case of Echinops sphaerocephalus different parts of the plant were investigated. The root and the stem are useful from a practical point of view because of the high uronic acid content. Some fractions, which seemed to be interesting on the basis of the measured uronic acid content, were dialyzed then concentrated, lyophilized and their uronic acid, protein and carbohydrate contents were determinated (Table III). The Echinops orientalis root fraction 6. was separated by ion-exchange chromatography. The monosaccharide composition of the neutral and the acidic fractions was measured by gas chromatography (Table IV). PMID- 9737022 TI - [Influence of auxiliary materials on the proportion of effective particle size of metered-dose suspension type aerosols]. AB - The aim of the present study was to formulate suspension type inhalation aerosols by various types of auxiliary materials, and to select the formulation with the highest proportion of the effective particle size. The examined suspension type aerosol contained sodium cromoglycate as an active compound. For the stabilization of the suspension, the applied surface active ingredients were oleic acid and oleyl oleate, and dimethyl siloxane polymer was selected as hydrophobizing agent. Factorial design was used for the optimization of the experimental results. On the basis of our results, the correct types and amounts of auxiliary materials can be selected to obtain the therapeutically effective formulation. PMID- 9737023 TI - [Experiences with the rectal use of a chemotherapeutic agent. 1. In vitro biopharmaceutical examinations; selection of the optimal vehicle]. AB - The factors influencing in vitro liberation (Part 1) and in vivo absorption (Part 2) from trimethoprim-containing rectal suppositories and the authors' results related to them are reported in this two-part publication. Special emphasis was laid on selecting the optimal suppository base which is harmless physiologically yet not indifferent pharmacologically. From among the 24 compositions studied, a lipophilic mixture containing a surface active additive (Witepsol W 35) and a hydrophilic (Macrogolum) mixture were found to be the best in all respects. Liberation from the trimethoprim-containing rectal suppositories was measured with in vitro dynamic diffusion and spectrophotometrically. A power relation was found to exist between the quantity of the released pharmacon and the diffusion time, and a significant negative exponential relation was observed between the doses and their respective in vitro availability values. PMID- 9737024 TI - [Experiences with the rectal use of chemotherapeutic agents. 2. Pharmacokinetic examinations with animals]. AB - The aim of the investigations was to optimise vehicle for trimethoprim (TMP) suppositories ready for clinical trials. The rectal absorption of TMP was studied in anaesthetized rats. The drug liberation properties of the five mixed vehicles with promising in vitro results (Part 1.) were studied. The course of the blood level curves was monitored with serial sampling. The TMP concentration of blood was determined by bioassay. Individual bases were compared with the use of the pharmacokinetic parameters derived from the analysis of the obtained blood level curves, with special respect to biological availability (BA). The extent of bioavailability is influenced considerably by the hydro-, lipo- or lipohydrophilic property of the vehicle. TMP, if incorporated in the proper vehicle, is absorbed well. With three vehicles the extent of absorption exceeded the absorption seen with oral administration on the same model (BA = 38.8%). The best results were achieved with the lipophilic base Witepsol W 35 containing 10% of Polysorbate 20 and 10% of Polysorbate 61 (BA = 63.8%) and with Witepsol W 35 containing 10% of Polysorbate 60 (BA = 64.9%). The hydrophilic Macrogol 1540 vehicle containing 5% of Macrogol 400 had only slightly worse results (BA = 52.9%). In the case of the lipohydrophilic Witepsol W 35 vehicle with 10% of Polysorbate 20 and 10% of Polysorbate 61 content a significant negative exponential relation was found between the administered doses and their respective bioavailability values, this tendency had been observed during the in vitro examinations, too. No such relation was found in the case of the lipophilic Witepsol W 35 vehicle containing 10% of Miglyol 812. PMID- 9737025 TI - [Review of bioanalytic methods in pharmacokinetics]. AB - The analytical methods used in pharmacokinetic practice are reviewed. The system of special pharmacokinetic demands on bioanalytical methods for carrying out the different pharmacokinetic, metabolite kinetic, bioequivalence and food interaction studies is demonstrated in details (high sensitivity; the parallel measurability of the parent compound and its metabolites; species and biological matrices dependent methods; anticoagulation; sample preparation; derivatisation; special calibration system; method selectivity; reproducibility; validation; economicalness). The different immunological methods and separation techniques used in pharmacokinetic practice and the sensitivity of the detectors have been compared and characterized. Without the requirement of completeness, the author has tried to summarize the most important bioanalytical techniques in the view of general and specific feature; the possibilities of application, characteristics of operation and limitations. PMID- 9737026 TI - [Determination of enantiomeric purity by simultaneous dual circular dichroism and ultraviolet spectrophotometry]. AB - A method is described for the determination of enantiomeric composition. The ellipticity and absorbances of the sample are measured simultaneously by CD and UV spectroscopies, and the resulting G value is determined. G is an intensive physico-chemical parameter, a close derivative of anisotropy factor. Its magnitude is identical with opposite sign for enantiomers. The experimental G value is concentration-independent, as long as both absorption and ellipticity are linear functions of concentration. The analytical procedure introduced here is simple, rapid, and inexpensive, even though it includes calibration with standards of established enantiomeric composition. Provided the sample contains some UV-active contaminant(s), the method can be used after achiral chromatographic purification. By virtue, the method lacks several sources of error, arising normally from concentration inaccuracies. Applicability of the principles is tested by the example of phenylglycine and mandelic acid. Advantages of the method allowed the determination of phenylglycine enantiomer purity with an accuracy of 0.1%. PMID- 9737027 TI - Deliberately caused bodily damage. AB - Meaning has the power to drastically modify our experience of pain. It also can dramatically affect our health, both positively and negatively--a phenomenon I explored previously in this column. Spiritual meanings may be the most powerful forms of meaning, because they can provoke healing responses that appear miraculous, such as in DCBD. DCBD bridges medicine, neuroscience, religion, and anthropology. Consequently, it is regarded as too sprawling by scientists who prefer to work in a confined, limited area. DCBD also carries the stigma of being "weird" and "foreign." So it isn't surprising that researchers have given it a wide berth over the years. At long last, however, this attitude is changing, and a few brave investigators are coming forward. In my imagination I see DCBD as a territory marked by bold signs: ATTENTION PROSPECTIVE NOBLE PRIZE WINNERS: LOOK HERE! Is anybody out there up to the challenge? PMID- 9737028 TI - The Grail's poetry ritual theatre: ancient healing for breast cancer survivors. PMID- 9737029 TI - Homeopathic and psychiatric perspectives on grief. AB - OBJECTIVE: This review describes the homeopathic analysis of grief and common remedies corresponding to this reaction. Homeopathic descriptions of grief are compared with contemporary psychiatric criteria. DATA SOURCES: Each homeopathic rubric (i.e., symptom) is identified on the basis of a computerized repertory search, grouped according to body systems, and compared with a current set of operational criteria derived from the psychiatric literature. The major homeopathic remedies for grief are identified. STUDY SELECTION: One hundred four rubrics for grief were found, incorporating mental and physical symptoms as well as physical disease. DATA SYNTHESIS: Homeopathic phenomenology of grief was closely matched with its current psychiatric definition. A close correspondence was seen between psychiatry and homeopathy, even though each has a differing heritage and temporal origin. The correspondence of a later descriptive system (i.e., psychiatry) to an earlier, independently derived system (i.e., homeopathy) confers validation to both systems' description of the grief response. CONCLUSION: The similarities and differences between homeopathic and psychiatric descriptions of grief have been noted. Similar forms of grief response are recognized by both systems, though homeopathy provides a more extensive list of physical sequelae following bereavement. Controlled trials of homeopathy in grief states are recommended. PMID- 9737030 TI - Clinical outcome research in complementary and alternative medicine: an overview of experimental design and analysis. AB - This article serves as a primer for those beginning clinical research in complementary and alternative medicine. The authors provide a basic overview of important experimental design and statistical issues, of which clinical researchers in the area of complementary and alternative medicine must be aware when attempting to demonstrate the effectiveness of particular treatment modalities. As the article suggests, science is an inferential process, and experimental investigations can vary greatly in methodological integrity. Key concepts in clinical outcome research such as internal validity, statistical conclusion validity, and the appropriate measurement and operational definitions of outcomes are discussed. New scientific approaches that are evolving because of paradigm shifts in science (e.g., chaos theory) are also reviewed. Suggestions are provided to further develop an understanding of clinical outcome research methodology. PMID- 9737031 TI - Heart-focused attention and heart-brain synchronization: energetic and physiological mechanisms. AB - CONTEXT: Many relaxation, meditation, and imagery techniques that implicitly or explicitly involve focused attention on the body, including qigong, massage, and noncontact therapeutic touch, purportedly employ energetic and physiological mechanisms. OBJECTIVE: To show that, from a perspective of dynamical energy systems, relaxed self-attention enhances connectivity between the brain and body. This enhanced connectivity may be achieved by at least 2 mechanisms: (1) physiological mechanisms employing peripheral negative feedback loops, and (2) bioelectromagnetic mechanisms involving direct energetic resonance between the peripheral organ and the brain. DESIGN: 19 channels of electroencephalogram, 1 electrocardiogram, and 2 channels of electro-oculogram were recorded from 22 subjects who focused their attention on their heartbeats or eye movements, with and without kinesthetic (touch) biofeedback to increase somatic awareness. RESULTS: Analyses of the electroencephalogram synchronized with the electrocardiogram revealed significant effects for heart-focused attention, primarily with touch biofeedback, following the contraction of the ventricles (possibly reflecting increased baroreceptor and somatosensory feedback); and significant effects for heart-focused attention, with and without touch biofeedback, preceding the contraction of the ventricles (possibly reflecting direct electromagnetic interactions between the heart and the brain). CONCLUSIONS: These findings suggest that energetic and physiological mechanisms may be involved in techniques in which the goal is to promote mind-body integration and health. PMID- 9737033 TI - Stephen G. Wright, MBE, FRCN creating sacred space. Interview by Bonnie Horrigan. PMID- 9737032 TI - A comparative study of chiropractic and medical education. AB - BACKGROUND: Chiropractic is the largest of the alternative/complementary health professions in North America. However, little attention has been given in the health sciences literature to the formal curriculum of chiropractic education or to its similarities to and differences from the curriculum of allopathic medical education. This lack of information precludes extensive referrals and interaction between the 2 professions, even when historical and political barriers can be overcome. METHOD: This is a descriptive, comparative study of the curriculum content of North American chiropractic and medical colleges, supplemented by in depth data obtained through site visits with 6 institutions (3 chiropractic and 3 medical). DISCUSSION: Considerable commonality exists between chiropractic and medical programs. Regarding the basic sciences, these programs are more similar than dissimilar, both in the types of subjects offered and in the time allotted to each subject. The programs also share some common areas in the clinical sciences. Chiropractic and allopathic medicine differ the greatest in clinical practice, which in medical school far exceeds that in chiropractic school. The therapies that chiropractic and medical students learn are distinct from one another, and the settings in which students receive clinical training are different and isolated from one another. With these similarities and differences established, future studies should examine the quality of the 2 educational programs in detail. PMID- 9737034 TI - Dreams as nonlocal connections. PMID- 9737035 TI - Previously published research may help to deflate TT controversy. PMID- 9737036 TI - Lack of control group deemed problematic in fibromyalgia pilot study. PMID- 9737037 TI - Reader defends other mysterious phenomena. PMID- 9737038 TI - Acupuncture treatment for phantom limb pain. PMID- 9737039 TI - Understanding vulvodynia. AB - Vulvodynia is a difficult management problem. In this review article, the clinical subsets of vulvodynia including recurrent vulvovaginal candidiasis, vulvar vestibulitis syndrome and dysaesthetic vulvodynia are described. Their aetiology is discussed and available therapies are presented. PMID- 9737040 TI - Hirsutes. II: Treatment. AB - The treatment of hirsutes includes cosmetic measures, such as bleaching, plucking, electrical epilation and, more recently, laser epilation. Pharmacological therapy consists of anti-androgens and includes the androgen receptor blockers spironolactone and cyproterone acetate. Other drugs reducing androgen expression include oral contraceptives and corticosteroids. A new follicular 5 alpha reductase inhibitor, finasteride, is currently under evaluation, as is the anti-androgen flutamide. Gonadotrophin-releasing hormone agonists reduce androgen expression and show early promise in the treatment of hirsutes. PMID- 9737041 TI - Botulinum toxin for the correction of hyperkinetic facial lines. AB - The present article illustrates the effects of low dose botulinum toxin (BTx) injections for the improvement of hyperkinetic facial lines and presents a grading treatment chart designed to standardize the reporting of the improvement seen. A questionnaire of patient acceptance, the patients' impression of therapy and short-term results and complications are reported. Twelve patients with 26 injected-paired regions were charted and the response to injection was graded. Patients had hyperkinetic facial lines in glabella, periorbital regions or horizontal forehead lines. Diluted BTx type A (1 IU/0.1 mL) was injected and patients were assessed at 10 days. A second follow up injection was offered to patients at this stage if required. Objectively, all patients' hyperkinetic actions and lines improved or diminished. The degree of improvement was similar in all areas injected and a symmetry of results was always observed. In a minority of cases, all movement was lost (7/26) and in others it was weakened but present (19/26). In some injected areas the actual expression line that was visible at rest disappeared entirely (11/26): in the others it was diminished (15/26). Complications were few. Two patients had temporary brow ptosis that spontaneously recovered within the first week. No eyelid ptosis was noted. Bruising and headaches were the most common reported complications. Low dose BTx is an effective and well-tolerated treatment for hyperkinetic facial lines with few significant complications in this small pilot study. The grading chart may allow easier comparisons of results between studies on the effects of BTx therapy. PMID- 9737042 TI - Occupational contact dermatitis among New Zealand farmers. AB - Forty-six farmers were patch tested to determine whether their dermatitis was secondary to an occupational allergen. Twenty-eight had a positive patch test of which 23 were thought relevant (definite or probable). In 20 of these cases, the allergen(s) was considered to be work related (define or probable). The common allergens were pesticides (N-(1,1,2,2-tetrachloroethylthio) -4-cyclohexene-1,2 dicarboximide (captafol), ethylenebis (dithiocarbamato) manganese (maneb) and copper sulfate), rubber compounds (N-isopropyl-N-phenyl-4-phenylenediamine (IPPD) and 4-phenylene diamine base) and sunscreen chemicals. PMID- 9737043 TI - Brachioradial pruritus and cervical spine manipulation. AB - Brachioradial pruritus (BRP) causes significant morbidity in the majority of patients for whom no effective treatment is found. Chronic ultraviolet radiation exposure has usually been cited as the cause, but nerve damage from cervical spine disease has also been implicated. We report on a small retrospective exploratory study, conducted by questionnaire, of a group of patients who were treated with a specific cervical spine manipulation. Ten of 14 patients reported resolution of symptoms following manipulative treatment. All six patients who had had previous cervical spine disease responded to manipulation, as did half the remaining eight patients who had no previous history of neck symptoms. Although patients with BRP, by definition, share similar symptoms, the aetiology is almost certainly multifactorial. Prospective studies looking for cervical spine disease, as well as assessment of this particular method of cervical spine manipulation as a treatment modality for BRP, should be considered. PMID- 9737044 TI - Hypersensitivity syndrome reaction to oral terbinafine. AB - Terbinafine is used extensively to treat onychomycosis and other dermatomycoses. The case of a patient who developed a hypersensitivity syndrome reaction to oral terbinafine is discussed. A 66-year-old male was placed on terbinafine (250 mg/day) for the treatment of onychomycosis. After 4.5 weeks of therapy, the patient developed a cutaneous eruption, pyrexia, lymph-adenopathy and hepatic dysfunction. No infectious or other cause was found for the symptoms and signs, which resolved within 6 weeks of stopping terbinafine. The patient had been on prednisone, doxazosin mesylate and aspirin for several months prior to starting terbinafine. These medications were continued during the episode and subsequently afterwards, with adjustment to the prednisone dosage only. The hypersensitivity syndrome reaction in this case involved multiple systems and was idiosyncratic in nature with no apparent predisposing factors. With the increasing use of oral terbinafine, it is likely that rare adverse events will occur more frequently. It is, therefore, important for physicians to be aware of the possible development of a hypersensitivity syndrome reaction in a patient on terbinafine who experiences an adverse event with multisystem involvement. Prompt recognition and determination of the extent of systemic involvement is important for the proper management of the patient. PMID- 9737045 TI - Mycobacterium marinum: chronic and extensive infections of the lower limbs in south Pacific islanders. AB - We report three adult cases of very chronic, extensive infection of the lower limbs due to Mycobacterium marinum. The patients were from South Pacific islands and, clinically, the widespread warty plaques resembled chromomycosis. One was associated with severe lymphoedema. All three patients gave a history of at least 20 years duration. The patients were otherwise well and not immunologically compromised. In all cases, the organism was identified on tissue cultures and was not seen on histopathology. The mycobacteria were sensitive to most antibiotics tested in vitro. The patients were treated with a combination of rifampicin and cotrimoxazole with good results. PMID- 9737046 TI - Benign lichenoid keratosis due to constant pressure. AB - Two well-circumscribed keratotic plaques in an elderly man, one each on symmetrically identical locations over the sides of the buttocks, are described. Clinical and histopathological features supported the diagnosis of benign lichenoid keratosis, which had probably resulted from constant pressure over the site and regressed when this factor was eliminated. PMID- 9737047 TI - Erythropoietic protoporphyria treated with narrow-band (TL-01) UVB phototherapy. AB - Erythropoietic protoporphyria is a rare photodermatosis for which treatment options are limited. The present report describes the clinical features of a patient with erythropoietic protoporphyria and liver function test abnormalities associated with treatment with beta-carotene. Subsequent treatment with narrow band UVB phototherapy resulted in marked subjective improvement in photosensitivity, which was confirmed by abolition of demonstrated abnormalities on monochromator phototesting. The therapeutic options for photosensitivity in erythropoietic protoporphyria are reviewed and discussed. PMID- 9737048 TI - Inoculation cutaneous tuberculosis. AB - Two cases of inoculation cutaneous tuberculosis are presented. As commonly occurs, the diagnosis could not be confirmed bacteriologically due to the small numbers of organisms present. However, both patients responded to antituberculosis chemotherapy. PMID- 9737049 TI - Axillary granular parakeratosis. AB - A 54-year-old woman had a 3 year history of a recurrent bilateral axillary rash during the summer months. Both axillae showed hyperkeratotic, fissured and cobblestone plaques. Skin biopsy showed the histology previously defined as axillary granular parakeratosis. This finding may indeed represent an unusual contact reaction to anti-perspirants interfering with epidermal keratinization. PMID- 9737050 TI - Novelty shop 'itching powder'. AB - To evaluate causes of itch, commercial 'itching powders' were sought for evaluation. Only one product, produced in Germany and consisting of ground rose hips, is currently sold in novelty shops in the Boston area. These plant fibres appear to provoke itch and prickle sensations by non-allergic mechanical stimulation, similar to the action of wool fibres. PMID- 9737051 TI - Innervated musculocutaneous lip flap (Karapandzic technique). AB - The Karapandzic technique is easy to perform and provides excellent results for full thickness defects larger than one-third of the lower lip. The flap is slid and rotated into position while an intact neurovascular pedicle is maintained. Accordingly, sensation and circulation of the lip is preserved and function of the orbicularis oris muscle is maintained. PMID- 9737052 TI - Australian aborigines and ringworm (tinea). PMID- 9737053 TI - Neuropathic pruritus. PMID- 9737054 TI - Response to the article, 'Pellagra in a woman using alternative remedies'. PMID- 9737055 TI - Primin sensitivity in a patient sensitive to Streptocarpus. PMID- 9737056 TI - The role of trauma and dissociation in cognitive-behavioral psychotherapy outcome and maintenance for panic disorder with agoraphobia. AB - The relationship between traumatic experiences and dissociation with pretreatment psychopathology and rates of recovery, relapse and maintenance for patients receiving cognitive-behavioral treatments for panic disorder with agoraphobia (PDA) were investigated. One-hundred and forty-seven subjects who met DSM-III criteria for agoraphobia with panic attacks and who completed participation in one of two previously conducted treatment outcome studies were mailed packets containing measures to assess history of trauma, victimization and dissociation. Eighty-nine of these were returned and completed sufficiently to be included in the present study. It was hypothesized that a variety of trauma-related variables (e.g. history of traumatic experience, type of trauma, age at which the trauma first occurred, perceived responsibility, social supports available, self perceived severity, level of violence, and whether or not the traumatic event was followed by self-injurious or suicidal thoughts and/or behaviors) and dissociative symptomatology would be predictive of (1) greater psychopathology at pretreatment, (2) poorer treatment response and (3) higher relapse rates and poorer maintenance over a 1 year longitudinal follow-up. These hypotheses were supported by the findings and the theoretical, empirical and clinical implications are discussed. PMID- 9737057 TI - Anger management style and the prediction of treatment outcome among male and female chronic pain patients. AB - Anger is a prominent emotion experienced by chronic pain patients. Anecdotes suggest that anger predicts poor outcome following multidisciplinary pain programs, but no empirical evidence documents this link. We expected that patient anger expression or suppression would predict poor outcome following a pain program and that gender differences would emerge. Pre- to posttreatment measures of lifting capacity, walking endurance, depression, pain severity and activity level were collected from 101 chronic pain patients. An 'anger expression x gender' interaction was found such that anger expression among males was correlated negatively with lifting capacity improvements. 'Anger suppression x gender' interactions emerged such that anger suppression among males was correlated negatively with improvements in depression and general activities. These effects remained significant after controlling for trait anger. Thus, how anger is managed may exert unique influence on outcomes apart from the effects of mere anger proneness, at least among male pain patients. PMID- 9737058 TI - Illusory correlation and social anxiety. AB - An illusory correlation (IC) experiment examined the presence of a phobia relevant covariation bias in the context of social anxiety. Low (n = 28) and high (n = 32) social anxious women were shown a series of slides comprising pictures of angry, happy and neutral faces which were randomly paired with either a shock, a siren or nothing. One half of the participants were shown women faces, whereas the other half were shown men faces. Participants indicated outcome expectancies on a trial by trial basis. After the experiment proper they estimated the contingencies of all slide/outcome combinations. Participants showed both an a priori and an a posteriori IC between angry faces and shock. This covariation bias was similar for men and women faces and independent of prior fear. The pattern of results is consistent with the idea that ICs arise from initial expectancies that survive extinction. PMID- 9737059 TI - Traumatic intrusions as 'worse case scenario's'. AB - While some clinicians assume that traumatic intrusions are historically accurate revisualizations of traumatic incidents, others have suggested that these types of intrusions may represent a worse case scenario (i.e. exaggerated) version of the trauma. To explore this issue, a survey was conducted among undergraduate students (N = 189). Of the 69 respondents who had been the victim of or witness to a relatively recent trauma, 15 (22%) reported an exaggerated perception of the traumatic incident. Exaggerated intrusions were found to have more flashback qualities and tended to have a higher frequency than 'realistic' intrusions. These findings are well in line with the idea that intrusions are not necessarily veridical copies of traumatic events. PMID- 9737060 TI - The convergent validity of the Phobia Origins Questionnaire (POQ): a review of the evidence. AB - The Phobic Origins Questionnaire (POQ) [Ost, L.-G. & Hugdahl, K. (1981). Acquisition of phobias and anxiety response patterns in clinical patients. Behaviour Research and Therapy, 19, 439-447.] is the most commonly cited instrument for determining the origins of phobic anxiety and data obtained using this instrument strongly support the role of conditioning in the acquisition of fear reactions. The construct validity of the POQ in assessing episodes of conditioning has been questioned [e.g. Menzies, R. G. & Clarke, J. C. (1994). Retrospective studies of the origins of phobias: a review. Anxiety, Stress and Coping, 7, 305-318.] This paper examined the convergent validity of the POQ by comparing origins' classifications based on the POQ to classifications based on alternative instruments. The convergent validity of the POQ was found to be extremely poor. The POQ was consistently associated with a much greater likelihood of classifying the origin of fear reactions as due to direct conditioning episodes than was found using alternative instruments. The findings question the usefulness of the POQ in examining the origins of phobic anxiety. PMID- 9737061 TI - A confirmatory factor analysis of posttraumatic stress symptoms. AB - Investigators have recently identified a two-factor structure underlying posttraumatic stress symptoms through the use of exploratory factor analysis. [Taylor et al. (1988). The structure of posttraumatic stress symptoms. Journal of Abnormal Psychology, 107, 154-160]. These two factors, which were labeled as Intrusion and Avoidance, and Hyperarousal and Numbing, are consistent with current theoretical models of posttraumatic stress disorder--PTSD [e.g. Foa et al. (1992). Uncontrollability and unpredictability in post-traumatic stress disorder: An animal model. Psychological Bulletin, 112, 218-238]. However, the authors of the Taylor et al. study issued caution in interpreting their findings because there has yet to be a systematic replication of their results. This paper presents a confirmatory factor analysis of the two-factor structure of posttraumatic stress symptoms in 217 survivors of serious motor vehicle accidents with varying degrees of PTSD symptoms. A hierarchical, confirmatory-factor analysis conducted with a structural equation modeling statistics package confirmed that the model proposed by Taylor et al. can adequately account for the presentation of PTSD symptoms in this sample of motor vehicle accident survivors. The implications for the assessment and diagnosis of PTSD are discussed. PMID- 9737062 TI - An outbreak of infection with Shigella flexneri 1b in south east England. PMID- 9737063 TI - Influenza immunisation to include all those aged 75 years and over. PMID- 9737064 TI - Increase in gonorrhoea cases in Grimsby. PMID- 9737065 TI - [Risk factors and correlations with some tumor markers in the incidence of breast cancer in pre- and post-menopausal patients]. AB - Between January and November 1992, forty eight women with benign disorders of the breast were studied and compared with one hundred fifty two patients with malignant disease of the breast. The content of alkaline phosphatase (ALP) levels was measured in the serum of premenopausal and postmenopausal patients. In all of them clinical history was taken, weight size and corporal mass index (CMI) (weight/size2). When the patients were grouped according to type of breast lesions, it appeared that plasmatic ALP levels, were higher in the groups with infiltrating carcinoma than in the women with benign disorders of the breast. The physiological status of the enzyme in this study, relate probably the activity of the tissue in either benign or malignant tissue, through fibroadenoma had less activity than adenocarcinoma. Status ALP (> or = 90 UI/L), CMI, menstrual state and histological type were compared. We found a significant correlation between these parameters. PMID- 9737066 TI - [Relation between body mass index and bone mineral density in a sample population of Mexican women]. AB - The purpose of this trial is to demonstrate that a women with high body mass index (BMI > or = 28) has greater bone mineral density (BMD) from that with lower BMI. We studied 922 healthy women who met the inclusion criteria. They were classified into four groups according to their BMI (> or = 28 and < 28) and age (> or = 35 and < 35 years). Bone mineral measurement was performed by dual-energy X-ray absorptiometry (DEXA) in the hip and at the lumbar region. BMD in overweight women older than 35 years was significantly higher in comparison with that of women with lower BMI, both in the hip and the lumbar spine. In overweight women younger than 35 years, we found greater BMD in the hip reaching statistical significance, but not at the lumbar spine. We conclude that obesity is associated with greater BMD (4% at the lumbar spine; 11% at the hip) probably due to both greater physical stress and higher estrogen levels. PMID- 9737067 TI - [A murine model of polycystic ovaries: its use in the evaluation of effects of endoscopic ovulation induction]. AB - The objective was to evaluate the effect of the endoscopic treatment of the polycystic ovary syndrome (PCOS) in a murine model induced with depot estrogen. Three groups with PCOS were studied: group 1 (n = 22) evaluated exclusively using laparoscopy; group 2 (n = 11) one fulguration on both ovaries was performed during laparoscopy; and group 3 (n = 10) three fulgurations were performed. One month after laparoscopy the animal were sacrificed and was carried surgical microscopic examination of the intraabdominal organs, moreover histological evaluation of the gonads was practiced. All the animals showed PCOS. No adhesions were observed in the animals of group 1, while the frequency of adhesion in group 2 was 36.3%, 18.1% of the animals of group 2 showed gonadal atrophy. The frequency of adhesions in the group 3 was 66.6%. The adhesion score was significantly higher in groups 2 and 3 than in group 1. Is concluded that the use of estrogen is useful to induce PCOS in animal models. The procedure can be employed to study the effects of the surgical induction of ovulation, it's utility in other conditions seems promissary. PMID- 9737068 TI - [Risk of transmission of infectious diseases by transfusion]. AB - In obstetric patients transfusion is a common procedure, it has many advantages but it also has severe risks. Since the observation that human immunodeficiency virus (HIV) infection is transmitted by transfusion, the number of preventive measures to reduce the infectious diseases transmission by this procedure has increased. The microorganisms that can be transmitted through transfusion include: human T lymphotropic virus (HTLV) I and II, hepatitis B virus, hepatitis C virus, hepatitis D virus, hepatitis G virus, HIV, cytomegalovirus, Treponema pallidum, Barucella sp, Toxoplasma gondii, Plasmodium sp, and Trypanosoma cruzi. The most important measure for reduce transfusion risks is the appropriate and careful use of this procedure. This article review transfusion's indication, describe the infectious diseases commonest transmitted by transfusion and analyze the preventive measures to put in practice. PMID- 9737069 TI - [Internal hirudiniasis: vaginal bleeding resulting from leech bite]. AB - A postmenopausic woman referred continuous bleeding during seven days, which suddenly started after she was swimming in the river. She didn't have a general symptoms. A gynecological examination showed that the cause of the bleeding was a leech bite on the vaginal wall. The worm was gave away by means of surgical forceps without complications. We described the clinical and pathological features of this unusual parasite attack. PMID- 9737070 TI - [Abdominal pregnancy, institutional experience]. AB - Abdominal pregnancy is a rare entity, which has been classified as primary or secondary by Studiford criteria. A retrospective study, between January 1989 and December 1994, realized at Instituto Nacional de Perinatologia, found 35,080 pregnancies, from which 149 happened to be ectopic, and 6 of them were abdominal. All patients belonged to a low income society class, age between 24 and 35 years, and average of gestations in 2.6. Gestational age varied from 15 weeks to 32.2 weeks having only one delivery at term with satisfactory postnatal evolution. One patient had a recurrent abdominal pregnancy, with genital Tb as a conditional factor. Time of hospitalization varied from 4 to 5 days, and no further patient complications were reported. Fetal loss was estimated in 83.4%. Abdominal pregnancy is often the sequence of a tubarian ectopic pregnancy an when present, it has a very high maternal mortality reported in world literature, not found in this study. The stated frequency of abdominal pregnancy is from 1 of each 3372, up to 1 in every 10,200 deliveries, reporting in the study 1 abdominal pregnancy in 5846 deliveries. The study had two characteristic entities one, the recurrence and two, the delivery at term of one newborn. Abdominal pregnancy accounts for 4% of all ectopic pregnancies. Clinical findings in abdominal pregnancies are pain, transvaginal bleeding and amenorrea, being the cardinal signs of ectopic pregnancy. PMID- 9737071 TI - [Intra-vesical translocation of an intrauterine device, report of a case]. AB - The intra-uterine device TCu 380 (IUD) has a great acceptance among the group of women that take the fertility control in not definitive form and it is being accepted by more women. In spite of the simple technique to insert the IUD, the people in charge of the planning family program in the first level attention section of the centers of health, have not enough knowledge for its insertion. We reported the case of a 21 year old woman that applied the IUD immediately after her baby's birth and she had sudden expulsion the following week. She went to the first level attention section where they inserted the IUD again and four months later she began to have urinary symptoms and abdominal pain, she went for a check, but could not find the filaments of the IUD. The x-ray of her abdomen showed an inverted IUD, they tried to take it off, but without any success. She was sent to the General Hospital where made an ultrasound that showed the IUD in the urinary bladder. A transurethral endoscopy was made and also an ultrasonographic dragging without gerring it. That is why decided to take it off by abdominal and urinary bladder surgery. The IUD was inserted transurethral which the diameter of the applier is thin and lets it go free in thorough the urethra, besides that during the operation did not find any harm in the uterus or the bladder that could make suspect the perforation in order to make the IUD reach the urinary bladder. The above mention demonstrate the ignorance of the technique and the anatomic place for the insertion of the IUD. That is the reason why it is necessary a more detailed preparation it the whole personnel assigned to the family planning programs. PMID- 9737072 TI - [True value of ultrasound in gynecology]. AB - With the purpose to determine the certainty of diagnosis of the most frequent gynecological pathology, susceptible of surgical treatment urgent or programmed, we studied 104 cases during a periods of nine months. In each case, we performed a clinical diagnosis, by ultrasound and surgery. Finally, we compared them with the histopathological diagnosis, obtaining the sensibility, the specification, and the predictive value for each one the methods and the most often pathologies. PMID- 9737073 TI - [Perinatal mortality at the Medical Care Units of the IMSS (Mexico Social Security Institute), National Medical Center of Torreon]. AB - To describe the situation of perinatal mortality during 1994 year in General Hospitals with Family Medicine number 16 and 18 of IMSS (Social Security Mexican Institut) National Medical Center in Torreon Coah. It was realized a retrospective study, were included 199 files of perinatal deaths occurred from January 1st to December 31 of 1994. The variables obtained were number of death for step, period, age, sex, weight and the cause of the cause of the death. Were eliminated the files without data of interest. For the analysis our utilized descriptive statistics. The rate of perinatal mortality was 20.17 per 1000 live birth, fetal death rate 9.58 by 1000 and the rate of neonatal death 12.97 per 1,000 live birth, fetal death rate 8.68 and neonatal death rate 12.30. The majority were in the perinatal period one with a rate of 16.71 by 1,000 live birth. Were most common in a male sex (53%) in pregnancies from 28 to 32 weeks (33.91%) and in babies with less of 1000 gr of weight (33.86%). The causes more frequents of deaths were the respiratory difficult syndrome (41.77%), the anomalies (19.62%) and hypoxia (9.49%). The perinatal mortality in our study was similar that in the rest of the country and is acorde with the literature. The perinatal mortality were in the perinatal period one. Is important to conduce a prospective studies. PMID- 9737074 TI - [Uniform requirements for the preparation of manuscripts sent to biomedical periodicals. International Committee of Editors of Biomedical Journals]. PMID- 9737075 TI - The future of self-regulation. PMID- 9737076 TI - Paraformaldehyde-containing paste in endodontic therapy. PMID- 9737077 TI - Disability claim review shows there's no claims crisis. PMID- 9737078 TI - The truth about fee guides. PMID- 9737079 TI - Chronic mandibular dislocation: the role of non-surgical and surgical treatment. AB - Although the management of acute dislocations of the temporomandibular joint (TMJ) has not changed significantly in recent years, chronic dislocations continue to be treated by a variety of methods. Long-standing cases are the most difficult and frustrating to manage. This paper reports on four cases demonstrating the signs and symptoms associated with some forms of chronic. TMJ dislocations, and the difficulties encountered in the management of some of these conditions. An algorithm based on a critical review of the literature is proposed for the management of both acute and chronic TMJ conditions, and recommendations are made on how to eliminate or reduce their recurrence. PMID- 9737080 TI - Antimicrobial resistance: dentistry's role. AB - The resistance of bacteria, fungi and viruses to antimicrobials is increasing rapidly, with deleterious consequences. Dentistry's role in this development is unclear, because the necessary information has not yet been collected. Nevertheless, dentists should recognize that it is essential to use antimicrobials in an appropriate and responsible manner, both to treat infection effectively, and to minimize the likelihood that the bacteria in the general population will develop resistance to antimicrobials. The purpose of this article is to make dentists aware of the concerns raised by antimicrobial resistance, and how it can be avoided. PMID- 9737081 TI - Resin-metal bonding systems: a review of the Silicoating and Kevloc systems. AB - The Silicoating and Kevloc systems are resin-metal attachment devices that are designed to improve bond strength and decrease microleakage at the resin-metal interface. Clinically, these systems can be used to place resin pontics and veneers, enhance cementation with resin cements, and bond the resin matrix to metal implant substructures. To provide practitioners contemplating the use of either modality with background information, a concise summary of the potential advantages of resin-metal bonding, the procedures required to establish the chemical bond, and the methods of evaluating laboratory studies are presented. Chemical bonding with these systems is technique sensitive. How the system is used may be as important to the final clinical results as is the choice of system. PMID- 9737082 TI - Antimicrobial treatment options in the management of odontogenic infections. AB - Most acute orofacial infections are of odontogenic origin. In normal hosts, however, they usually do not occur without some type of predisposing condition. Early recognition and management of acute orofacial infections is critical, because rapid systemic involvement can occur, especially in children. Antimicrobial therapy has an essential role in the management of these infections. If it is initiated before surgery, it can shorten the period of infection and minimize associated risks. The etiology of odontogenic infections is usually attributed to the endogenous flora of the mouth, and not to the introduction of non-resident bacteria. Odontogenic infections are typically polymicrobial; however, anaerobes generally outnumber aerobes by at least four fold. The penicillins have historically been used as the first-line therapy in these cases, but increasing rates of resistance have lowered their usefulness. Bacterial resistance to this class of agents is predominately achieved through the production of beta-lactamases. Clindamycin, because of its broad spectrum of activity and resistance to beta-lactamase degradation, is an attractive first line therapy in the treatment of odontogenic infections. PMID- 9737083 TI - Coping with divorce. PMID- 9737084 TI - The TMD controversies--a second opinion. PMID- 9737085 TI - Clinical research in the Department of Veterans Affairs: using research to improve patient outcomes. AB - This brief description of a few VA clinical research activities does not do the program justice but does provide insight concerning priorities and prospects for future clinical research. In addition to doing clinical research, the VA has renewed its commitment to the training and career development of clinical investigators, especially physician investigators. We have had an excellent response to our solicitations for career development awards for medical, surgical, and psychiatric junior faculty who wish to pursue a clinical research career. The continued collaboration between VA and the academic community is critical to this revitalization of our research career development programs. Clinical research short-courses and didactic programs are also available to our investigators through similar collaborative relationships. The future is promising for clinical research in the Department of Veterans Affairs. We enjoy strong support from the Department leadership, Veterans Service Organizations, and many policy makers in Washington who support health related research activities year after year. We work hard to justify their support of and confidence in our abilities to improve health care for the many ill veterans who benefit from our research. PMID- 9737086 TI - The J.I.M. interview. Purnell W. Choppin, MD. AB - The Howard Hughes Medical Institute (HHMI) is known worldwide for its support of high calibre research in the basic biomedical sciences. At present, 332 men and women at 72 institutions are HHMI investigators, putting them in the ranks of the best and the brightest (and certainly the most generously supported) researchers in America. But the Howard Hughes Medical Institute's portfolio goes well beyond its core program as a medical research organization. Purnell W. Choppin, MD, discussed the breadth of Hughes' activities during a recent interview in his office at the Institute's main campus in Chevy Chase, Maryland, a few blocks away from the National Institutes of Health. Among other things, Choppin talked about US medical schools, clinical research, postgraduate education, and HHMI support of research abroad. PMID- 9737087 TI - Bootstraps and jackknives: limitations. PMID- 9737088 TI - Macrophage inflammatory protein-2 predicts acute lung injury in endotoxemia. AB - BACKGROUND: Proinflammatory mediators that include tumor necrosis factor-alpha (TNF-alpha) and macrophage inflammatory protein-2 (MIP-2) and anti-inflammatory mediators such as interleukin-10 (IL-10) modulate the immune response to endotoxemia. IL-10 downregulates the production of TNF-alpha and MIP-2. Acute lung injury may occur secondary to neutrophil chemotaxis mediated by chemokine MIP-2. We studied the temporal relationship of TNF-alpha, MIP-2, and IL-10 in rat endotoxemia and correlation of MIP-2 concentrations with acute lung injury. METHODS: Ten ventilated rats were randomized to receive an intravenous infusion of 2 mg/kg Escherichia coli lipopolysaccharide (n = 6) or saline placebo (n = 4). Blood pressure was continuously monitored and arterial blood was obtained for lactate, blood gas, TNF-alpha, IL-10, and MIP-2 measurements at baseline, 2, 4, and 5.5 hours after LPS or saline infusion. RESULTS: Endotoxemia resulted in hypotension, lactic acidemia, and increased alveolar-arterial oxygen gradient (A a O2 gradient) compared with the placebo group. TNF-alpha, MIP-2, and IL-10 levels were increased 2 hours after endotoxemia. Subsequently, TNF-alpha levels declined while IL-10 and MIP-2 levels remained elevated. Control rats had no significant increase in cytokine production at any time point. MIP-2 concentrations correlated with A-a O2 gradient, an indicator of lung injury (r = 0.56, p < 0.001). CONCLUSIONS: MIP-2, possibly released by TNF-alpha stimulation of macrophages, is associated with acute lung injury possibly by inducing neutrophil chemotaxis. IL-10 may exert its counter-inflammatory response by inhibiting the release of TNF-alpha in endotoxemia. PMID- 9737089 TI - Evidence for selective degradation of platelet GP IIb/IIIa complex after prolonged in vitro ventricular assist. AB - BACKGROUND: Left ventricular assist devices (VAD) have improved survival in patients with end-stage heart failure. Past studies have shown that interactions between blood and synthetic surfaces promote initial bleeding and later thromboembolism. The exact mechanism of blood activation during VAD circulation remains unclear. The purpose of this study was to test the hypothesis that platelet glycoprotein (GP) IIb/IIIa receptor degradation occurs during clinical use of ventricular assist devices. METHODS: Five in vitro nonpulsatile centrifugal VAD circuits were simulated for 4 days using 450 mL of fresh human whole blood. Temperature, activated clotting time, pH, pCO2, pO2, Ca++, and glucose were maintained at physiologic values. Flow was maintained at a constant 2.0 L/min/m2. We examined whole blood platelet aggregation induced by ristocetin, collagen, and adenosine diphosphate (ATP). We also examined whole blood platelet degranulation induced by collagen and ADP. RESULTS: Platelet aggregation in response to ristocetin, collagen, and ADP irreversibly and progressively declined with prolonged circulation in the VAD. While ADP-induced aggregation declined within the first hour, ristocetin and collagen-induced aggregation declined after 10 hours. Collagen-induced platelet degranulation decreased similar to aggregation, whereas ADP-induced degranulation continued and was preserved throughout the experiment. CONCLUSIONS: These results suggest prolonged circulation of human blood in a VAD circuit irreversibly impair platelet aggregation. The response of circulating platelets to individual agonists suggests that this platelet degradation is partially receptor specific. In our VAD system, ADP-stimulated platelet aggregation is more rapidly degraded with circulation. These results offer preliminary evidence that circulation of human blood in a VAD circuit leads to early degradation of the platelet GP IIb/IIIa complex. GP IIb/IIIa complex degradation is likely to be the mechanism of early VAD associated bleeding. PMID- 9737090 TI - Enhanced expression of the cytoskeletal-associated protein, paxillin, in experimental nephrotic syndrome. AB - BACKGROUND: In the model of aminonucleoside of Puromycin (PAN)-induced nephrotic syndrome we assessed changes in glomerular expression of three proteins that regulate cell adhesion to extracellular matrix: paxillin, focal adhesion kinase (FAK), and Rho. METHODS: Following a single intravenous injection of PAN in Sprague-Dawley rats, proteinuria ensued and glomeruli were isolated at three stages: prior to onset of proteinuria (days 1 and 2), and when proteinuria peaked (day 9), subsided (day 29) or resolved (day 35). Glomerular protein lysates were analyzed by Western blot for expression of paxillin, FAK, and Rho. RESULTS: There was a progressive increase in glomerular paxillin level that peaked concomitantly with heavy proteinuria (day 9). Paxillin remained increased during the recovery phase of PAN-induced injury and when proteinuria resolved. Expression of FAK and Rho remained unchanged at all time points. To explore whether the increase in paxillin expression following administration of PAN was due to a direct effect on glomerular epithelial cells (GEC), cultured rat GECs were incubated with PAN for 3, 6, and 24 hours, and expression of paxillin was assessed in GEC lysates by Western blot analysis. No change in paxillin levels was observed. CONCLUSIONS: In PAN-induced nephrotic syndrome there is a preferential increase in paxillin expression that cannot be accouted for by an effect of PAN on GEC paxillin synthesis. We propose that the enhanced paxillin synthesis in the course of PAN induced GEC injury reflects perturbations in contact between GEC and the GBM and may play a role in regulating adherence of GEC to the GBM. PMID- 9737091 TI - Effect of selective inhibition of the inducible cyclooxygenase on renin release in healthy volunteers. AB - BACKGROUND: Nonsteroidal anti-inflammatory drugs (NSAIDs) can block renin release via inhibition of cyclooxygenase (COX). The responsible COX-isoenzyme in man is unknown. Therefore, we assessed the effects of meloxicam, a selective inhibitor of COX-2, and indomethacin, an unselective inhibitor of COX-1 and COX-2, on furosemide stimulated plasma renin activity (PRA). METHODS: In a randomized cross over design 15 healthy female volunteers received no NSAID or meloxicam 7.5 mg/d for 6 days or indomethacin 25 mg tid for 2 days and 25 mg on the 3rd day. On the control day and on the last day of each treatment the following parameters were evaluated before and after furosemide 20 mg i.v.: PRA measured by RIA, urinary excretion of prostaglandin E2 (PGE2) assessed by gas chromatography-tandem mass spectrometry, urine volume and urinary excretion of sodium, potassium, and creatinine, and serum concentrations of sodium, potassium, and creatinine. RESULTS: Furosemide led to a more than two-fold rise of PRA. Indomethacin as well as meloxicam had no significant effect on basal PRA but inhibited the furosemide stimulated PRA increase. PGE2 excretion on the control day rose two-fold after furosemide. Meloxicam had no effect on basal PGE2 excretion, whereas indomethacin reduced this parameter by 30%. Both drugs inhibited the increase of urinary PGE2 after furosemide. No drug effects on urine flow nor on electrolytes and creatinine in serum and urine could be observed. CONCLUSION: Meloxicam inhibited furosemide stimulated renin release, suggesting that in man COX-2 is responsible for prostaglandin synthesis mediating renin release. PMID- 9737092 TI - Enhanced proliferation, apoptosis, and matrix accumulation by mesangial cells derived from HIV-1 transgenic mice. AB - BACKGROUND: Mice, transgenic for HIV-1 genes, have been demonstrated to develop renal lesions mimicking HIV-associated nephropathy. Focal glomerulosclerosis (FGS) has been reported to be the predominant glomerular lesion in these animals. In the other models of FGS, the accumulation of mesangial matrix and mesangial cell proliferation have been shown to be the preceding abnormalities. We evaluated the proliferation, apoptosis, and matrix accumulation by mesangial cells derived from mice transgenic for HIV-1 genes as well as from nontransgenic mice. METHODS: Mesangial cells were cultured from mice transgenic for HIV-1 genes (HTrMC) and nontransgenic mice (NTrMC) of the same age and sex. The growth rate of HTrMC and NTrMC was determined under identical conditions. Morphologic evaluation of apoptosis was performed by staining cells with Hoechst (H)-33342 and propidium iodide. Accumulation of mesangial cell collagen type IV, laminin, and fibronectin was measured by the dot blot assay. Total RNA was extracted from HTrMC and NTrMC and Northern blots were generated. These blots were probed with specific probes for TGF-beta, proteoglycan (P16), and GAPDH. RESULTS: Mesangial cells (HTrMC) derived from transgenic mice had greater (P < 0.004) proliferation when compared to mesangial cells (NTrMCs) from nontransgenic mice (HTrMCs, 4.2 +/ 0.3 vs NTrMCs, 3.0 +/- 0.2 x 10(4) cells/well). HTrMCs also showed enhanced (P < 0.0001) apoptosis compared to NTrMCs (HTrMCs, 13.2 +/- 1.5% vs NTrMCs, 3.1 +/- 0.5% apoptotic cells/field). HTrMCs accumulated an increased (P < 0.02) amount of collagen type IV (HTrMCs, 5659.7 +/- 472.8 vs NTrMCs, 3882.2 +/- 339.7 ng/well); whereas NTrMCs accumulated a greater amount of laminin when compared to HTrMCs (HTrMCs, 12.8 vs NTrMCs, 29.6 +/- 2.9 ng/well). HTrMCs also showed an enhanced mRNA expression of TGF-beta and an attenuated expression of proteoglycan (P16). CONCLUSIONS: These results suggest that mesangial cells derived from mice transgenic for HIV-1 genes have enhanced proliferation and collagen accumulation. The enhanced expression of TGF-beta may have contributed to enhanced HTrMC proliferation and the accumulation of collagen. The present study provides the basis for a hypothesis that mesangial cells may be contributing to the development of focal glomerulosclerosis in mice transgenic for HIV-1 genes. PMID- 9737093 TI - In vivo engraftment potential of clinical hematopoietic grafts. AB - BACKGROUND: Quantitative assays are needed to characterize the multilineage engraftment potential of clinical hematopoietic grafts. After we observed a dose response relationship between the number of human hematopoietic cells transplanted into nonobese diabetic-scid/scid (NOD/SCID) mice and the number of human CD45+ cells recovered in the chimeras' marrows and spleens, we sought to develop a multiple linear regression model that allows quantitative comparisons of human cell engraftment in vivo. METHODS: We used this NOD/SCID xenotransplant model to compare the engraftment potential of cord blood vs. adult marrow or mobilized blood, after either of 2 commonly used clinical graft engineering procedures: CD34+ cell selection or T cell depletion. RESULTS: The engraftment per transplanted cell was > 20 fold higher for cord blood cells, as compared to hematopoietic cells from adults. However, there was no difference in engraftment per CD34+ cell transplanted between marrow and mobilized blood. Levels of human cell engraftment from all sources could be increased by administration of human hematopoietic growth factors to human/mouse chimeras after transplantation. Correlation analysis of the number of human CD13+ myeloid cells and CD19+ B lymphoid cells in the chimeras' marrows 8 weeks after transplantation provided evidence that almost all the human myeloid and B lymphoid cells were derived from the same primitive precursor cells. CONCLUSIONS: These findings and assay may be useful in the development of clinical hematopoietic cell therapies. PMID- 9737094 TI - Hemodynamic response to vasopressin in dehydrated human subjects. AB - BACKGROUND: Despite the known potent vasoconstrictor effects of vasopressin, the role of this hormone in the maintenance of blood pressure is incompletely understood. In studies performed in animals with increased plasma vasopressin concentrations, several complex cardiovascular effects have been noted, including decreases in heart rate and cardiac output, which may account for a lack of effect on arterial pressure despite the vasopressin-induced increase in total peripheral resistance. Only a few studies have been done to assess the cardiovascular effects of vasopressin in human subjects, and most of these have been limited to measurement of heart rate and arterial pressure only. The present study was designed to identify more fully the cardiovascular effects of vasopressin when plasma vasopressin concentrations are increased by osmotic stimulation without the superimposition of major nonosmotic stimuli associated with severe volume depletion. METHOD: Studies were performed on 11 normal human subjects in supine and erect posture before and after 24 hours of fluid deprivation, and following administration of a selective V1 receptor antagonist, [d(CH2)5Tyr(ME)]AVP, after dehydration. Cardiovascular parameters were measured noninvasively by thoracic electrical bioimpedance cardiography and blood samples for measurements of plasma concentrations of vasopressin and other hormones affected by dehydration and differences in posture were collected for subsequent analysis. RESULTS: After 24 hours of fluid restriction, plasma osmolality was increased from 287 +/- 0.9 to 294 +/- 0.7 mosm/kg H20 and plasma vasopressin concentrations (Pavp) were increased in both supine and erect posture. Mean arterial (MAP) and systolic blood pressure (SBP) were reduced by fluid restriction but were higher in erect than in supine posture both before and after fluid restriction. Heart rate (HR), diastolic blood pressure (DBP), and systemic vascular resistance (SVRI) were also higher in erect than in supine posture, while cardiac index (CI), stroke index (SI), end-diastolic index (EDI), and an index of total thoracic fluid content (TFC) were all reduced in erect posture, both before and after dehydration. Plasma renin activity (PRA) and plasma norepinephrine concentrations (Pne) were increased in erect posture, both before and after dehydration, but there was no effect of erect posture on plasma vasopressin concentrations (Pavp), either before or after dehydration. Administration of the V1 receptor antagonist after dehydration had no effect on hemodynamic parameters other than small reductions in DBP and cardiac preload. CONCLUSION: It is concluded from these studies that small increases in Pavp associated with moderate dehydration do not play a role in the maintenance of arterial pressure in normal human subjects in either supine or erect posture. PMID- 9737095 TI - Pain relief and sclerosis of bone metastases in a patient with breast cancer treated with tamoxifen, radiotherapy and pamidronate disodium: which treatment helped? PMID- 9737096 TI - Nicotine withdrawal as an etiologic factor in delirium. PMID- 9737097 TI - Can tumors act as opioid traps, mimicking opioid tolerance? PMID- 9737098 TI - Assessing children's state anxiety. AB - Anxiety is an important component of children's pain and is routinely assessed in pain research. Two forms of the State-Trait Anxiety Inventory have been used frequently by researchers investigating children's pain and state anxiety (form C 1 and Y-1). We were unable to find psychometric information about this tool when used with a population of hospitalized children. Therefore, we undertook to assess reliability and validity, and identify problem items using data from 881 hospitalized children (aged 5-18 years) whom we had tested. Considering results of all analyses together, we concluded that the tools lack validity and reliability, and contain many problem items that are in need of revision. PMID- 9737099 TI - Development and psychometric assessment of the physical symptom distress scale. AB - While the importance of subjective symptom distress for clinical evaluation of end-stage renal disease (ESRD) is generally acknowledged, an adequate method to classify and quantify this distress is currently unavailable. The purpose of this study was to develop the Physical Symptom Distress Scale (PSDS) to assess patients' physical symptom distress accompanying ESRD. The sample consisted of 160 ESRD patients from three dialysis centers of hospitals in Taipei, Taiwan. Internal consistency reliability of the instrument was found to be good: alpha coefficient = 0.87 for the entire scale, and alpha coefficients = 0.79 for each subscale, respectively. Test-retest correlation of 0.82 with a 2-week interval supported stability reliability. Factor analyses indicated and confirmed "Fluid and electrolyte imbalance" and "Disturbance in neuromuscular function" domains. Support for concurrent validity was provided by correlation (r = -0.46) between the entire scale and the Karnofsky Performance Scale (KPS). For Factor I the correlation was -0.51, for Factor II the correlation was -0.33. The predictive validity of the PSDS was supported through multiple regression. These findings suggest that the PSDS is a reliable and valid measure of symptom distress for ESRD patients treated with hemodialysis. PMID- 9737100 TI - A survey of practice in management of malignant ascites. AB - The purpose of this study was to determine physicians' attitudes toward and preferences for palliative management of malignant ascites. A random sample of eighty physicians practicing in Canada was selected from the memberships of the Canadian Association of Medical Oncologists, the Canadian Association of Gastroenterology, the Canadian Society of Palliative Care Physicians, and the Society of Gynecologic Oncologists of Canada. Physicians were questioned on their use of different modalities in management of malignant ascites, and preferences based on attitudes toward efficacy of various treatments. Eighty surveys were mailed, with a second mailing, followed by telephone contact. The response rate was 76% (59/78), with two potential respondents deemed ineligible. Among the 44 physicians who treat malignant ascites, paracentesis is employed by 43 (98%), and felt to be effective by 39 (89%). Diuretics are used by 61% (27/44), although fewer feel diuretics are effective management (20/44, 45%). Peritoneovenous shunts, dietary measures, and other modalities are used less frequently than either paracentesis or diuretics. The most commonly used means of managing malignant ascites is paracentesis, which is also felt to be the most effective by the group surveyed. After paracentesis, diuretics and peritoneovenous shunting are used most frequently, but there is no apparent consensus as to their effectiveness. Managing malignant ascites remains problematic, and we propose further study of management strategies to clarify the role of various treatments. PMID- 9737101 TI - A clinical evaluation of transdermal therapeutic system fentanyl for the treatment of cancer pain. AB - Fentanyl has been incorporated into a transdermal therapeutic system (TTS) containing a rate-limiting membrane that provides constant release of the opioid. TTS fentanyl provides continuous opioid delivery for up to 72 hr without the need for special equipment. After Institutional Review Board approval, 53 patients with cancer pain requiring 45 mg or more of oral morphine daily were admitted into an open-label, prospective, multicenter evaluation of TTS fentanyl for the relief of pain. After a 1-week stabilization on oral morphine, patients were transferred from morphine to an appropriate dose of TTS-fentanyl (25, 50, 75, or 100 micrograms/hr) administered as a transdermal patch every 3 days. TTS fentanyl was titrated to pain relief, and patients were followed up for as long as 3 months. Pain relief and the side effects of the medications were assessed daily. Twenty-six men and 27 women with a mean (+/- SD) age of 61 (+/- 12) years entered the study; 23 patients completed the full 84-day study. The mean duration of TTS fentanyl use was 58 +/- 32 days. The mean (+/- SEM) daily morphine dose during the last 2 days of stabilization was 189 (+/- 20) mg, and the mean initial fentanyl patch dose was 58 (+/- 6) micrograms/hr. The mean daily morphine dose taken "as needed" for breakthrough pain at study completion was 35 mg. The mean final fentanyl dosage at study completion was 169 (+/- 29) micrograms/hr. Pain relief was rated as good or excellent by 82% of patients during the treatment period. When asked to compare pain relief during the first month of TTS-fentanyl use to that provided by their last analgesic before study entry, 63% preferred TTS fentanyl. Side effects considered related to the fentanyl patch were nausea (13%), vomiting (8%), skin rash (8%), and drowsiness (4%). Thirty percent of patients reported adverse experiences related to the fentanyl patch, and 17% had to be discontinued from the study. We conclude that TTS fentanyl administered every 3 days for the treatment of cancer pain is effective, safe, and well tolerated by most patients. PMID- 9737102 TI - Neuropsychological and pharmacokinetic assessment of hospice inpatients receiving morphine. AB - Eighteen inpatients receiving morphine for cancer pain in a palliative care unit were recruited to a study employing a range of neuropsychological tests to assess cognitive function. The tests employed were National Adult Reading Test, Williams Delayed Recall Test, Immediate Memory for Digits, Trailing Making Test, and the Digit Symbol Substitution Test. These data were correlated with biochemical tests of renal and hepatic function, morphine dose, route of administration, plasma morphine, morphine-3-glucuronide (M3G) and morphine-6-glucuronide (M6G) concentrations. Despite having no clinical evidence of impairment of cognitive function, the level of current intellectual functioning (Symbol Digit Substitution Test) was on average two standard deviations below normal. Immediate memory appeared to be well preserved, but Delayed Recall and Trailing Making Test scores were significantly above normal. There was no significant correlation between morphine dose, or plasma morphine and M3G and M6G concentrations, and the neuropsychological test results, although a weak correlation was found between plasma morphine concentration and digits forward (r = -0.47, p < 0.05) and Digit Symbol Substitution scores (r = -0.46, p < 0.05). Seven patients had some degree of nausea or vomiting, ascribed as an opioid adverse effect, and had higher serum creatinine concentrations, worse neuropsychological performance, and significantly higher plasma M3G concentrations (p < 0.05). These data provide some evidence to suggest that cognitive functioning in patients with advanced cancer receiving morphine may be significantly impaired despite apparent clinical normality. From these data it is not possible to determine what relative causal contribution the disease and the drug made to these observations, although renal function, plasma morphine, and M3G concentrations may be important. Future research should address a broad range of neuropsychological testing to assist in the modification of practices aimed at enhancement of quality of life, such as opioid substitution or rotation. PMID- 9737103 TI - Paroxetine for pruritus in advanced cancer. AB - Pruritus associated with malignancy may be one of the most bothersome symptoms in advanced cancer. Its control is still difficult to achieve and is a challenge to palliative medicine specialists. We describe five patients suffering from pruritus of different etiologies who responded rapidly to administration of paroxetine, a serotonin reuptake inhibitor, in a dose-dependent manner. Two patients experienced transient but severe nausea and vomiting. We suggest that paroxetine's antipruritic effect may be explained by rapid downregulation of the 5-HTs receptors, which may have an important role in the generation of pruritus and pain. PMID- 9737104 TI - Baclofen, a treatment for chronic hiccup. AB - The efficacy of baclofen in the treatment of chronic hiccup is demonstrated in two cases. These cases highlight the present state of knowledge related to hiccup. This discussion focuses on the definition and classification of hiccup, etiologies, postulated theories to explain its function, the few studies performed to date, and non-pharmacologic and pharmacologic treatment. Baclofen appears to be the agent most efficacious in the treatment of chronic hiccup. Its commonest side effect is sedation; insomnia, dizziness, weakness, ataxia, and confusion also can occur. Following regular use, abrupt discontinuation can lead to withdrawal symptoms, such as seizure, and gradual discontinuation is recommended. PMID- 9737105 TI - Case presentation: believing that your injection killed the patient. PMID- 9737106 TI - Commentary: a nursing perspective. PMID- 9737107 TI - Commentary: tensions between the theory and practice of palliative care. PMID- 9737108 TI - Comparison of the fertilization rate after intracytoplasmic sperm injection (ICSI) using ejaculated sperms, epididymal sperms and testicular sperms. AB - Intracytoplasmic sperm injection (ICSI) has been successfully used to achieve fertilization and pregnancies for patients with extreme oligoastheno-zoospermia using ejaculated sperms or patients with azoospermia using epididymal or testicular sperms. The aim of this study was to compare the fertilization rate after ICSI using ejaculated, epididymal and testicular sperms. Between January and September 1997, 10 azoospermic men underwent percutaneous epididymal sperm aspiration (PESA) or testicular sperm extraction (TESE) to recover sperm for ICSI. A total of 5 PESA cases and 5 TESE cases were performed at the Center for Assisted Reproduction & Embryology. Thirty-one patients performed ICSI using ejaculated sperms during the same period of time were used as a control group. ICSI using ejaculated sperms, epididymal sperms from PESA and testicular sperms from TESE was a highly successful technique, achieving fertilization rates of 78.5 per cent, 83.3 per cent and 80.8 per cent, respectively. Good fertilization rates were achieved without significant differences among the various sperm sources. PMID- 9737109 TI - Effectiveness and cost analysis of community-based rehabilitation service in Bangkok. AB - This paper aimed to clarify the effectiveness and the cost of the community-based rehabilitation service in Klong Toey slum after a three year study period. One hundred and seventy eight patients used community-based rehabilitation during the three year period. One hundred and fifty-seven patients (86.5%) reported that their problems/conditions were cured or improved. Only nine patients (5.1%) reported that they stopped using community-based rehabilitation because their problems/conditions did not improve. A statistically significant improvement in pain level and walking velocity assessment, in 105 and 78 patients respectively, was demonstrated. Total cost and cost per patient-day of the community-based rehabilitation were Bt 559,920 and Bt 111.1 respectively. Cost per-patient-day of this community-based rehabilitation service was compared with an estimated cost per patient-day of using rehabilitation services at Chulalongkorn University Hospital and was found to be cheaper. This study supported the role of community based rehabilitation in Thailand. The need for a health service study in rural areas was also noted. PMID- 9737110 TI - Clinical management and outcome of eclampsia at Rajavithi Hospital. AB - The purpose of this clinical study was to review experience in the management, and outcome of eclamptic patients at Rajavithi Hospital. Standardized treatment for all cases of eclampsia has consisted of magnesium sulfate intravenously and intramuscularly to control convulsions by means of Chesley and Tepper's regimen, intravenous hydralazine intermittently to lower diastolic blood pressure when it exceeds 110 mmHg, and initiation of delivery as soon as the patient has regained consciousness and is stable. During a ten-year period there were 167,200 deliveries and 90 eclamptic patients, yielding an incidence of eclampsia of 1 in 1,857 deliveries. There were three maternal deaths (3.3%) due to intracerebral hemorrhage. Serious adverse maternal outcomes were more frequent in women whose convulsions occurred before delivery. Excluding postpartum cases, perinatal mortality of fetuses weighing 1,000 g or more was 11.7 per cent. Magnesium sulfate is the drug of choice for treatment of eclamptic convulsions. In most situations, clinical assessment of deep tendon reflexes, respirations, and urine output is adequate to monitor maternal magnesium toxicity without the need to determine actual maternal serum magnesium levels. PMID- 9737111 TI - Choledochal cyst: review of 74 pediatric cases. AB - Seventy-four cases of CDC were treated at the Children's Hospital from 1977 to 1995. Female to male ratio was 5:1. Forty per cent of the patients developed symptoms within one year after birth and 75 per cent within 5 years of age. About one third of the cases were treated surgically within one year of age. Only 5 patients or 6.8 per cent had all the clinical triads of jaundice, abdominal pain and mass. Jaundice was the most common symptom in infants below one year of age but abdominal pain was the most common symptom in older children. Three newborn infants had associated biliary atresia. Established liver cirrhosis was noted during surgery in 9 patients who were operated upon within one year of age but none of the older children. All but one had either type I or type IV CDC, while the remaining one had Caroli's disease or type V CDC. Excision was the preferred treatment. Overall operative mortality rate after excision was 6.5 per cent. None of the patients who had internal drainage procedures succumbed. Infants below one year of age appeared to have high morbidity and mortality rates after surgery. Internal drainage procedure may be considered in high risk patients with advanced cirrhosis. Available information suggests that the occurrence of this disease in Thailand is probably more common than in Western countries and etiology of CDC in infants may be different from that in older children or adults. PMID- 9737112 TI - Thyroid scintigraphy in children with hypothyroidism: a five-year experience. AB - The presence or absence of thyroid glandular tissue demonstrated by thyroid scintigraphy is important for genetic and prognostic counseling and for acceleration of diagnosis in other affected siblings. Technetium-99m pertechnetate thyroid scintigraphy was performed on 27 children with cretinism at the Division of Nuclear Medicine, Faculty of Medicine Siriraj Hospital during the 5-year period from June 1991. Based on scintigraphic findings, three main groups of thyroid localization were seen. Thirteen (48.1%) were athyrotic while 3 (11.1%) had an ectopic thyroid and 11 (40.8%) had gland in normal position. Perchlorate discharge test was performed in 8 children of the last group and the results were positive indicating an organification defect. Thyroid scintigraphy and perchlorate discharge test provided the useful information for diagnosis, follow-up, and prognosis in children with cretinism. PMID- 9737113 TI - Comparison of short versus long duration of ampicillin and gentamicin for radical hysterectomy. AB - Prophylactic antibiotic therapy for radical hysterectomy is still controversial. Although the efficacy of antibiotics have been demonstrated, there remains the question of duration of administration. In this study, we retrospectively reviewed 95 patients who underwent radical hysterectomy and pelvic lymphadenectomy for cervical cancer at our institute. The management was uniform except for the duration of antibiotic administration. Group I (34 cases) had ampicillin and gentamicin for 3 days while group II (61 cases) had the same regimen for 7 days. No significant difference was found in terms of postoperative infection (2.9% in group I and 1.6% in group II) or febrile morbidity (32.4% versus 50.8%). Other factors such as the patients' age, body weight, preoperative hemoglobin level, amount of blood loss and blood transfused, operative time, duration of retroperitoneal drain and duration of suprapubic cystostomy. Only operative time had a significant influence on febrile morbidity regardless of the duration of antibiotics administered. In conclusion, the antibiotic administration gave a radical hysterectomy and pelvic lymphadenectomy a very low incidence of postoperative infection. Longer duration of treatment did not appear to lessen postoperative infection nor febrile morbidity. Shorter duration of antibiotic administration needs further evaluation. PMID- 9737114 TI - Experiment of intranasal synthetic filter for prevention of suspended particulated matter: rhinomanometric evaluation. AB - The experimental study of intranasal synthetic filter with stent was tried to decrease suspended particulate matter for human respiratory tract. Facial mask or surgical mask were evaluated. Nasal vestibular size in Thai adults was estimated. Different kinds of stents and filters were used. Standard anterior rhinomanometry was the proper objective method to test nasal air flow resistant of stent and filter. Nasal obstructive symptom correlated well with rhinomanometric results. One layer of outer and inner face mask at each end of the cylindrical silicone stent was the suitable device. There were no complications or side effects. This personal protective device was cheap and available. The filtration efficacy should be tested in a general population during a highly air-polluted period. PMID- 9737115 TI - Potassium contents of northeastern Thai foods. AB - From our previous nutritional assessment, low potassium (K) intake among northeastern Thai males has been clearly demonstrated. This prompted us to undertake a survey of the K content of local foods. Food samples comprised of 57 animal and 142 plant products which were collected from various places in the northeast of Thailand. The dry ashing method was used to prepare the samples for K analysis using an atomic absorption spectrophotometer. Foods could be divided into 7 groups according to their K levels. Foods containing K > or = 1000 mg per 100 g fresh food were categorized in group 1. These were mainly foods in the legume group, i.e., soybean, cowpea and mungbean. While rice (polished) and rice products, the main staple, were in group 7, the lowest K group of less than 100 mg per 100 g fresh food. Comparison studies of the natural foods between those collected from the northeast and from the central regions of the country, and between the cooked foods purchased from the rural villages and from the urban areas of Khon Kaen municipality, showed that, for most food items, the K content was similar wherever it came from. However, when the K content in various parts or in different stages of growth of the same kind of plants or animals was compared, a great variation was clearly seen, for example, young tamarind leaves contained K in group 6 whereas ripe tamarind fruit contained K in group 1. According to our food consumption data, the analysis of food components of 48 meals taken during the hot season by 13 rural volunteers revealed that food items eaten with the highest frequencies and in the largest amount were those in the low K food groups, i.e., glutinous rice (group 7) and green papaya (group 6). Our results suggest that the low K intake of these northeast rural Thai people is not due to a low K content of foods in this region, but rather that their food habits and low socioeconomic status restricts consumption of those food items with higher K contents. PMID- 9737116 TI - Preoperative portal vein embolization in major hepatectomy for hilar cholangiocarcinoma: a case report. AB - We herein, report a 48-year-old Thai man with underlying Child A cirrhosis from chronic hepatitis B who complained of right upper abdominal pain. The imaging studies revealed an incomplete obstruction of the hepatic duct confluence with intrahepatic bile duct dilatation, predominantly on the right side. Hilar cholangiocarcinoma Bismuth Type IIIa was considered to be the diagnosis. Portal embolization of the right portal vein was performed by transileocecal approach, combined liver and bile duct resection with bilio-enteric anastomosis was carried out three weeks later. The postoperative course was uneventful. We believe that portal embolization may benefit patients with hilar cholangiocarcinoma by decreasing postoperative liver failure. PMID- 9737117 TI - Anti Ro/SSA positive undifferentiated connective tissue disease in a mother with a newborn with complete congenital heart block: a case report. AB - An association between complete congenital heart block (CCHB) and anti-Ro/SSA antibody is well recognized but has never been reported in Thailand. We report here a 37-year-old female who was admitted because of massive epistaxis secondary to immune thrombocytopenia. She had given birth to a child with CCHB 2 years previously, when she was healthy. Antinuclear antibody and anti-Ro/SSA were positive in her sera, but were negative in her son. The relationship between anti Ro/SSA antibody and outcome of mothers with infants with CCHB is reviewed. PMID- 9737118 TI - Penicillium marneffei mesenteric lymphadenitis in human immunodeficiency virus infected children. AB - Disseminated P. marneffei infection is one of the common opportunistic infections seen in HIV-infected patients in Southeast Asia. We report 3 cases of HIV infected children with mesenteric lymphadenitis presented with prolonged fever and abdominal pain. The first two patients were diagnosed as peritonitis and acute appendicitis prior to exploratory laparotomy. Operative findings revealed multiple enlarged mesenteric lymph nodes. Histopathologic findings of mesenteric lymph nodes biopsy were characteristic for P. marneffei infection. Mesenteric lymphadenitis in the last patient was diagnosed by abdominal ultrasound. All three cases had positive blood and bone marrow cultures for P. marneffei. These patients were treated with amphotericin B. Fever declined in 3-6 days. The first two patients survived but the last one subsequently died from underlying hemophilia A (GI bleeding). CONCLUSION: Acute mesenteric lymphadenitis can be one of the unusual manifestations caused by P. marneffei. Southeast Asia is an endemic area for P. marneffei and is severely affected by acquired immunodeficiency syndrome epidemic. Therefore, mesenteric lymphadenitis should be considered in HIV-infected persons who present with prolonged fever and abdominal pain. PMID- 9737119 TI - Atypical granular cell tumor of the neurohypophysis: a case report with review of the literature. AB - A 76-year-old man had an atypical granular cell tumor of the neurohypophysis which showed pleomorphic nuclei, mitotic figures, and spindle-shaped cells, extremely rare findings to be encountered. Review of 45 patients with neurohypophyseal granular cell tumor revealed a ratio of 1:2 between male and female with the peak occurrence (31%) in the fifth decade, and with the mean age of 50 years. There were no patients below 20 years of age. The common clinical presentations included visual disturbances and endocrinopathies relating to sex hormones. Surgical removal was the treatment of choice. If it is possible, total extirpation should be attempted. Because of uncertain cellular origin, the lesion should be descriptively diagnosed as granular cell tumor although multiple terms have been proposed. PMID- 9737120 TI - Ethical decision-making and its teaching. PMID- 9737121 TI - Etiologic study of primary congenital hypothyroidism. AB - The underlying causes of 35 children with primary congenital hypothyroidism at the Children's Hospital were studied. There were 21 girls and 14 boys. Serum T4 and TSH level, 24 hours 131I uptake, and technetium-99m thyroid scintigraphy were performed after discontinuation of synthetic thyroid hormone for 4-6 weeks. Athyrosis was the most common cause and accounted for forty-three per cent of the patients. Twenty per cent of the patients had thyroid hypoplasia. Ectopic thyroid was found in thirty-three per cent of the patients. Only a patient whose diagnosis was organification defect had slightly enlarged thyroid gland, high retention of 131I and positive perchlorate discharge test. Onset of symptoms before 9 months of age may be helpful for distinguishing between lingual thyroid and the others. Serum T4 level less than 2 micrograms/dL was observed to be more common in athyrosis and lingual thyroid groups than thyroid hypoplasia group. PMID- 9737122 TI - Factors associated with state hospital utilisation among Thai elderly who had illnesses which needed hospitalisation. AB - Of the 4,480 elderly subjects in a multistage random sampling household survey of a National Survey of the Welfare of the Elderly in Thailand (SWET), 669 (14.9%) reported that they had been hospitalised during the last year and were recruited in an analysis which aimed to examine associated factors of state hospital utilisation among Thai elderly. Seventy eight per cent had been admitted once during the last year. Mean (standard deviation) duration of hospital stay during the last year was 11.9 (20.1) days. For the last period of hospitalisation, 532 elderly (79.5%) were admitted to state hospitals. One hundred and nineteen elderlies (17.8%) used private hospitals. Only 18 elderly (2.3%) used both state and private hospitals. According to the causes of hospitalisation, the elderly who used state hospitals were not more severely ill than those who used private hospitals. Nine univariate factors associated with state hospital utilisation were entered in a logistric regression model in which five independent determinants were identified including 'do not have electricity', 'heads of the family are not their children', 'do not have own savings', 'live in rural area', and 'have heard about free health care programme'. The Ministry of Public Health and organisations which are concerned with the elderly should allocate more resources to advertising a free health care programme for Thai elderly. PMID- 9737123 TI - Effect of dietary modification on changes in serum lipids among rural Thai persons with hypercholesterolemia. AB - This article reports on the effect of dietary modification on changes in eating patterns and serum lipids among hypercholesterolemic persons aged 40-59 years with no evidence of coronary heart disease in Mae Sot District, Tak Province, between 1995 and 1996. A total of 381 persons with total cholesterol levels > or = 240 mg/dl and triglyceride levels < 400 mg/dl were educated, counseled, and followed-up by the mobile health team at the health centres in the communities. The team comprised both hospital personnel (a physician, a health educator, and public health nurses) and the health centre workers. Of the 381 study persons, 331 (86.9%) completed the one-year follow-up. The participants at one-year follow up were more likely than at baseline to reduce intakes of dietary fat and cholesterol, whereas, there was an increased intake of vegetables and fruits. The mean total cholesterol level significantly decreased from 258.9 mg/dl at baseline to 236.1 mg/dl at one-year follow-up (p < 0.01), giving an 8.8 per cent reduction. The mean change in low-density lipoprotein cholesterol levels was a 26.0 mg/dl decrease (p < 0.01), yielding a 15.1 per cent fall. The mean high density lipoprotein cholesterol level increased from 44.6 mg/dl at baseline to 46.8 mg/dl at one-year follow-up (p < 0.01). The proportion of those who had a body mass index of < 25 slightly increased from 70.7 per cent at baseline to 72.5 per cent at one-year follow-up. The dietary intervention program by the mobile team may be useful for lowering serum cholesterol among the rural population with hypercholesterolemia. PMID- 9737124 TI - Impediment of the progressions of microalbuminuria and hyperlipidemia in normotensive type 2 diabetes by low-dose ramipril. AB - In a randomized, double-blind, placebo-controlled study, we investigated in normotensive type 2 diabetics with microalbuminuria the effect of ramipril, an ACE inhibitor, on urine albumin excretion and serum lipids. A total of 1,882 patients were screened for urine microalbumin consecutively by dipstick test, Rapi Tex-Albumin test and RIA. The final 28 normotensive and microalbuminuric patients were assigned to receive either ramipril (1.25 mg/d, n = 16) or placebo (n = 12) for 12 weeks. Throughout the study, both groups had no changes in blood pressure, fasting plasma glucose, HbA1C, serum creatinine and electrolytes and no difference in creatinine clearance. At week 12 only the placebo group showed the significant increment of urine albumin excretion and triacylglycerol (30.6 +/- 38.3 to 39.0 +/- 19.7 and 167 +/- 64 to 208 +/- 77 mg/dl, respectively) but the decrement of HDL-cholesterol (46 +/- 16 to 35 +/- 6 mg/dl). During a 3 month period, increased urine albumin excretion was observed in normotensive type 2 diabetes with microalbuminuria who received only placebo. We conclude that ramipril may arrest the progression of albumin excretion and had favorable effects on serum lipids. Ramipril was safe and well-tolerated without untoward side effects during the study period. PMID- 9737125 TI - Comparison of two-layer Percoll gradient and mini-Percoll methods for sperm preparation. AB - To compare the efficiency of sperm preparation between the two-layer Percoll gradient and mini-Percoll methods, 50 normal and 33 abnormal semen samples from male partners of infertile couples were studied. The number of recovered spermatozoa, percentage of motility, percentage of normal morphology, and their survival at 24 and 48 hours were assessed. Both Percoll gradient techniques resulted in a significantly higher percentage of motility and percentage of normal morphology compared with the original semen samples (p < 0.0001). The two layer Percoll gradient showed a higher sperm recovery than the mini-Percoll method (p < 0.001), but the latter resulted in a higher percentage of motility (p > 0.001) and a higher sperm survival rate at 24 hours (p < 0.05) than the former, regarding normal semen samples. These differences did not appear with abnormal semen samples when analyzed as a group. Considering each of the abnormal parameters separately, sperm recovery was significantly higher after the two layer Percoll gradient in the case of astheno- and teratozoospermia (p < 0.05), but sperm survival at 48 hours was higher after the mini-Percoll gradient in the case of teratozoospermia (p < 0.05). It is concluded that both the two-layer Percoll gradient and mini-Percoll method can be used effectively for sperm preparation. The former yields a higher sperm recovery, but the latter should be considered regarding teratozoospermic samples and semen samples of very low volume. PMID- 9737126 TI - Video thoracoscopic lung biopsy in diffuse interstitial lung diseases. AB - The present study examined the use of video thoracoscopic lung biopsy (VTLB) in diffuse interstitial lung disease, in comparison with open lung biopsy (OLB). Twenty and fifteen patients underwent VTLB and OLB, respectively, from 1987 to 1997 at the Central Chest Hospital, Thailand. Data in mean (SD). The mean age was 39 years in both groups. VTLB yielded equivalent size of lung tissues, 4.7 (2.32) cm3, and was as diagnostically useful as OLB. Estimated blood loss, 60 (37) mls, and length of pleural drainage, 2.8 (0.5) days, were comparable in either technique. As OLB had been in practice for decades, it took shorter operative time, 64 (11) mins, than VTLB, 105 (30) mins, (p = 0.005). Both VTLB and OLB approaches were safe and not associated with major postoperative complications. PMID- 9737127 TI - Effect of umbilical vein oxytocin injection on the third stage of labor: a randomized controlled study. AB - To evaluate the effect of umbilical vein oxytocin injection on the duration of third stage of labor, and estimated blood loss within 24 hours postpartum, in 50 vaginal parturients at Rajavithi Hospital from March 1, 1994 and June 30, 1995. The parturients were randomized to administered either an umbilical vein injection of 20 units of oxytocin diluted to 20 ml with normal saline (oxytocin group) or only normal saline 20 ml (control group) immediately after cord clamping. There were 25 paturients in each group. The duration of the third stage of labor in the oxytocin group was significantly shorter than that in the control group. In only 1 case of the control group was manual placental removal performed. The estimated blood loss within 24 hours postpartum in both groups was not statistically different. Twenty units of umbilical vein oxytocin injection was effective to shorten the duration of the third stage of labor but were not effective to reduce the estimated blood loss within 24 hours postpartum. The need for a further large scale study in the future was suggested. PMID- 9737128 TI - Prognostic importance of p53 and c-erbB-2 oncoproteins overexpression in patients with breast cancer. AB - Using immunohistochemistry, 119 breast cancer tissues were examined for overexpression of p53 and c-erbB-2 oncogene proteins. In 46 (38.7%) of the cases p53 was overexpressed, while 35 (29.4%) demonstrated positive c-erbB-2 immunostaining. Expression of these two oncogene products was closely correlated (p < 0.01). There was no significant association between p53 protein expression and age of the patients, clinical stage, tumor size, number of involved nodes or estrogen receptor status. However, we found significant correlation between p53 protein expression and 5-year disease-free survival (p = 0.0113). In addition, the findings in this study clearly indicated that the co-overexpression of p53 and c-erbB-2 proteins was a powerful predictor for early recurrence in the patients with breast cancer. PMID- 9737129 TI - The scarless rhytidectomy incision in superficial parotidectomy. AB - Post-operative scars usually occur at the incision sites following various procedures of the parotid gland surgery. From 1982 to 1996, 201 cases of scarless rhytidectomy incisions were analysed and reported, with satisfactory results. PMID- 9737130 TI - Solitary fibrous tumor arising from hyperplastic thymus. AB - One case of a solitary fibrous tumor arising from the hyperplastic thymus is recorded. The patient was a 37-year-old female who presented with an anterior mediastinal mass. Thoracotomy was performed and revealed that the tumor arose on a pedicle from the posteroinferior surface of the enlarged thymus. The pathologic findings were characteristic of a solitary fibrous tumor. This is a very rare neoplasm that occurred in the mediastinum and had evidence of thymus gland in origin. PMID- 9737131 TI - Epithelial-myoepithelial carcinoma of parotid gland: a case report with immunohistochemical and ultrastructural studies. AB - A 66-year-old man presented with a painless mass of the parotid gland. Light and electron microscopic studies verified the basic nature of the tumor as epithelial myoepithelial carcinoma, a low-grade malignant neoplasm of the salivary gland. Pathologically, there were two types of cells; the inner eosinophilic epithelial cells lining the ducts and the outer clear cells. The former cells displayed immunoreactivity for cytokeratin and ultrastructural features of apical microvilli and desmosome. The latter cells were positive for actin, S-100 protein, vimentin and the cytoplasm contained actin microfilaments. Such pathological findings were characteristic features of this rare tumor. To our knowledge, this is the first reported case of EMC in Thailand. PMID- 9737132 TI - Pentazocine-induced fibrous myopathy and localized neuropathy. AB - A 47 year-old woman who had a 4-year history of intramuscular pentazocine injections in the lower extremities, developed gradual stiffness and weakness of the lower extremities. The thigh and buttock muscles were "wooden-hard" on palpation. The skin was hard, shiny and hairless. Associated clinical and electrophysiological polyradiculopathy and multiple mononeuropathy of the lower extremities were observed. Imaging studies showed calcification and fibrosis of the involved muscles. Muscle biopsy revealed fibrous myopathy. Caution in longterm usage and early recognition of pentazocine toxicity as a neuromuscular complication are important in order to prevent irreversible drug-induced fibrous myopathy and localized neuropathy. PMID- 9737133 TI - Pulmonary dirofilariasis in a patient with multisystem Langerhans cell histiocytosis--the first reported case in Thailand. AB - Despite a high prevalence of canine dirofilariasis, there is no case of pulmonary dirofilariasis reported from Thailand. We herein report a case of multisystem Langerhans cell histiocytosis who had an incidental pulmonary dirofilariasis found at the time of autopsy as a solitary nodule at the periphery of the right lower lobe. This is the first reported case in Thailand. Association between pulmonary dirofilariasis and Langerhans cell histiocytosis has not been described before in the literature. PMID- 9737134 TI - 7 year-survival of 2 cases of multicentric or metastatic osteosarcoma. AB - Osteosarcoma is a fatal disease. Neoadjuvant chemotherapy combined with irradiation treatment provide a better survival. In the Faculty of Medicine, Ramathibodi Hospital, the overall 9 year survival probability was 55 per cent among 130 cases of more than Enneking stage II osteosarcoma. Between 15 cases of bony metastases, there were 2 cases which were classified as multifoci osteosarcoma or osteosarcomatosis. These two cases developed second bone disease, 32 and 38 months after initial diagnosis and survived for 84 and 88 months with one patient also developing pulmonary metastasis. Both of them are still alive and in very good health. PMID- 9737135 TI - Medical ethics and professionalism. PMID- 9737136 TI - Adaptation and evaluation of the Liverpool Seizure Severity Scale and Liverpool Quality of Life battery for American epilepsy patients. AB - The Liverpool Seizure Severity Scale (LSSS) and the Liverpool Quality of Life (LQOL) battery were developed in Great Britain to assess the severity of seizure symptoms and the impact of epilepsy on patients' quality of life. The scales have been validated on British patients, but have not been validated for use with American patients. The objectives of this study were to adapt the scales to the American population and to evaluate their reliability and validity. After modifications recommended by focus groups with patients and epilepsy specialists, the scales were administered to a sample of 90 epilepsy patients who had experienced seizures within the previous 4 weeks. Comparisons of patients with generalized tonic-clonic seizures (n = 58) and partial seizures (n = 32) revealed significant differences on 9 of the 20 items on the LSSS as well as the total score. None of the six LQOL subscales (negative drug effects, positive drug effects, affect balance, sense of mastery, life fulfilment and impact of epilepsy) distinguished patients with different seizure types but five of the six subscales were significantly correlated with seizure severity. The internal consistency and test-retest reliability were adequate for both the LSSS and LQOL. Finally, five of the six LQOL scales were significantly correlated with independent measures of mental health, physical health and role functioning. PMID- 9737137 TI - Testing for differences in multiple quality of life dimensions: generating hypotheses from the experience of hospital staff. AB - In clinical trials with a quality of life (QoL) component, it is usual to monitor several QoL dimensions at several points in time. Multiple significance tests without formal hypotheses are problematic. It is not always feasible to specify a priori hypotheses for all variables. Can such studies be used to generate hypotheses for testing in later research only? We developed a method which can allow for formal hypothesis testing on a data set collected without a priori hypotheses in the protocol. We surveyed experienced physicians and nurses treating patients, to obtain independent expectations about differences in QoL dimensions. These 'staff expectations' will be used in the analysis of QoL data collected from breast cancer patients taking part in three randomized trials of adjuvant therapy. We propose frameworks for the informal and formal use of the experience of the staff in testing for group differences in patients' QoL scores. The method described here is anticipated to be useful for QoL studies in general, even when a priori hypotheses were specified before the studies were initiated. PMID- 9737138 TI - Estimating the effect of treatment on quality of life in the presence of missing data due to drop-out and death. AB - Quality of life was measured in a study of two multidrug regimens for mycobacterium avium complex MAC bactaeremia using the MOS-HIV questionnaire. The effect of treatment on quality of life was estimated at each follow-up time in three ways: (1) using only the observed data, (2) after assigning the worst possible quality of life scores for individuals who died, and (3) after imputing missing scores for patients who either died or dropped out of the study. The overall quality of life scores were also compared between treatment groups with categorical generalized estimating equation models and three-dimensional graphs. Of the 179 patients included in these analyses, 84 (47%) either died or dropped out during the 16 week study period. When the quality of life scores were compared between the treatment groups with the Wilcoxon rank sum test using only the observed data, there was no significant difference between the groups at 16 weeks of follow-up. When the worst possible quality of life scores were assumed for patients who had died, both the magnitude and the statistical significance of the difference in the quality of life scores between the groups increased. Imputing missing data for patients who either dropped out or died resulted in even larger differences in quality of life between the treatment groups. We conclude that ignoring missing data due to drop-outs and death can result in an underestimation of the treatment effect and overly optimistic statements about the outcome of the participants on both treatment arms due to the selective drop out of participants with poorer quality of life. To obtain a valid assessment of the effect of treatment on quality of life, the experience of the patients who died or dropped out of the study must be considered. PMID- 9737139 TI - Psychological well-being among cancer patients receiving radiotherapy--a prospective study. AB - The impact of cancer on the psychological well-being of newly diagnosed cancer patients before and during the course of radiotherapy was assessed in 70 consecutive cancer patients. Most of the patients were over 40 years of age, women, illiterate and from a lower socioeconomic group. During the course of treatment there was a decrease in the well-being scores on some dimensions such as perceived family and primary group support. Improvements were seen in the dimensions of positive feelings, coping, social support other than the family and spiritual well-being. There were no changes in the dimensions of negative feelings and perceived ill-health. The results give a profile on well-being and the changes observed during radiotherapy. PMID- 9737140 TI - Parental quality of life and recurrent ENT infections in their children: development of a questionnaire. Rhinitis Survey Group. AB - During the first 4 years of life, children develop ear, nose and throat (ENT) infections because their immune system is not fully mature. Some of them will develop a recurrence and repetition of these episodes might have impact on the parents' quality of life. Based on a literature review, face to face interviews with parents and meetings with a paediatrician, a 17 item French questionnaire has been devised in order to evaluate the specific impairment of parents' quality of life during a winter season. Cultural adaptations were made for Italy, Germany, Portugal and the Czech Republic. This questionnaire was completed by 1,214 parents in a longitudinal survey. The questionnaire was reduced to 14 items and three scores were identified through principal component analysis: an emotional score (eight items), a daily disturbance score (six items) and a global score (14 items). Construct validity was confirmed through multitrait analysis and internal consistency reliability confirmed using Cronbach's alpha. The emotional score was found to be linked to the number and type of ENT episodes while the daily disturbance score was linked to social consequences. The global score was linked to both aspects, demonstrating the clinical validity. This questionnaire is therefore able to measure the cumulative effect of recurrence of children's ENT infections on their parents' quality of life in France, Italy, Germany, Portugal and the Czech Republic. PMID- 9737141 TI - Patients' experiences using a computerized program with a touch-sensitive video monitor for the assessment of health-related quality of life. AB - This study assessed patients' experiences using a computerized program with a touch-sensitive video monitor (TSVM) for the assessment of health-related quality of life (HRQoL). A software program was developed for a computerized form of the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire, the QLQ-C30. One hundred and seventy-eight patients completed the QLQ-C30, followed by a structured interview designed to assess perceived difficulties with the use of the TSVM. Patients were asked to evaluate the ease of use of different aspects of the TSVM system (using the touch-sensitive screen, entering the patient identification number, reading the screen and following the on-screen instructions). The majority of patients found all aspects of the TSVM system 'very easy' to use. A few patients (1-2%) admitted finding any aspect of the TSVM use 'somewhat difficult' and none ranked any aspect as 'very difficult'. There were no unanswered items in the QLQ-C30. All patients found the amount of time spent on answering the questionnaire acceptable, (the mean time to complete was 6.8 min with a median of 5 min) and 96% stated they were willing to complete a similar questionnaire on a future occasion. From the patients' perspective the TSVM system appears to be a highly acceptable approach for the collection of HRQoL data in clinical practice. PMID- 9737142 TI - The impact of Gaucher disease and its treatment on quality of life. AB - To obtain information about how Gaucher disease and its treatment, specifically enzyme replacement therapy, affect patients' health-related quality of life (HRQoL), we interviewed 16 patients with type I Gaucher disease (range 8-67 years). All but three patients had been receiving enzyme replacement therapy for at least 6 months. The quality of life factors examined for these patients included physical health, social life, emotional health, financial burden, future plans and satisfaction with health care. The results indicated that bone pain and chronic fatigue interfered with school, job and social activities and were the most debilitating symptoms of Gaucher disease. Most patients experienced a significant increase in energy level from therapy and reported significant improvements in quality of life. Most patients did not perceive an effect of Gaucher disease on their overall emotional health, but some patients expressed anxieties about the discomfort, inconvenience and high costs of therapy. We conclude that a measure of HRQoL for Gaucher patients should include a generic core of items supplemented by disease-specific items designed to assess the changes in symptoms and in the occupational, recreational, social and emotional aspects of patients' lives that occur as a result of disease progression and/or management. PMID- 9737143 TI - Health-related quality of life varies with personality types: a comparison among cancer patients, non-cancer patients and healthy individuals in a Japanese population. AB - In an attempt to examine differential effects of personality on health-related quality of life (HRQoL) without regard to disease type, we used the HRQoL-20, a general questionnaire (Japanese original scale) we developed (comprising 20 questions related to physiological, psychological or social HRQoL) and the Eysenck Personality Questionnaire (EPQ), which measures personality traits of extraversion (E), neuroticism (N) and psychoticism (P). The subjects (399 males and 429 females), stomach cancer patients, non-cancer patients (who had received acupuncture or moxibustion treatment) and healthy controls, were classified into three personality types. The results indicated that the HRQoL score of the tolerable/tolerant type (high E, low N and high P scorers) was greater than the intolerable/intolerant type (low E, high N and low P scorers) and also the unclassified type (neither of above scorers). The HRQoL correlated positively with the E and P scales and negatively with the N scale, in the case of all subjects, with the exception of N in male cancer patients and E in male non cancer patients. The results supported the hypothesis that the HRQoL varies with personality variables, in that each patient, in different treatment settings, strives for the situation that is congruent with his/her personality to attain a better HRQoL. PMID- 9737144 TI - The influence of decisions made by developers on health status questionnaire content. AB - To investigate the effect of stringent and lenient criteria upon the process of item selection in the development of a health status questionnaire, an item pool (179 items) was administered to 139 patients with left ventricular dysfunction. Associations between each item and the criteria of gender, age, duration of disease, global health and global impairment were examined. Items were selected from the pool on the basis of their associations with the criteria using four levels of stringency. The most stringent criteria rejected items which had a shared variance of > or = 4% with gender, age and duration of disease and a shared variance of > or = 6% with global health and impairment. The most lenient criteria rejected items which had a shared variance of > or = 6% with gender, age and duration of disease and a shared variance of > or = 4% with global health and impairment. Using the most stringent criteria, 75 items were selected, compared with 127 items using the most lenient criteria. Small differences in the level of association had large effects on item selection. The choice of level of association used to base item selection can have a crucial influence on questionnaire content. PMID- 9737145 TI - Discriminant validity of the Medical Outcomes Study cognitive function scale in HIV disease patients. AB - Human immunodeficiency virus (HIV) infection results in a chronic course of disease progression and eventual death. With this disease progression comes decreases in health-related quality of life and cognitive function in many patients. We evaluated the construct and discriminant validity of the Medical Outcomes Study four-item and six-item cognitive function scale in a sample of 162 patients with HIV disease. The patients were assessed at baseline and after 4 months with the cognitive function scale, a cognitive functioning transition item, the Reitan trail making test (TMT) and the Centers for Epidemiologic Studies Depression (CES-D) scale. The results found that the four- and six-item cognitive function scales varied by HIV disease stage, CD4 count, self-reported change in cognitive function, TMT-based cognitive impairment and depression. The differences in the cognitive function scores were attenuated, but still remained statistically significant even after controlling for depression. The cognitive function scales predict cognitive impairment based on the TMT. The results support the construct validity of these self-report cognitive function scales. PMID- 9737146 TI - [Brain abscesses]. PMID- 9737147 TI - [Brain abscess. Clinicomicrobiologic study and prognostic analysis of 59 cases]. AB - INTRODUCTION: Clinical, microbiological, therapeutic and prognostic characteristics of brain abscesses were analyzed as well as the influence of CT in their evolution. MATERIALS AND METHODS: Retrospective study of 59 patients with the diagnosis of brain abscess of bacterial source before (group A) and after (group B) the introduction of CT (25 and 34 patients, respectively). RESULTS: The most common symptom was headache (76.3%) and the most common abnormality in physical examination was a decrease in the level of consciousness (61%) and this abnormality was associated with a higher mortality rate (13% versus 41.6%; p < 0.05) and also a higher proportion of neurologic sequelae (50% versus 85.7%; p < 0.05). The diagnosis was obtained earlier in group B. The hematogenous source predominated (32.2%); an adjacent source was identified in 28.8% and an apparent source was not recognized in 27.2% (40% in group A versus 17.6% in group B). Anaerobic and microaerophilic streptococci were the bacteria recovered most frequently. Gram-negative aerobic bacteria were the most common in otogenic abscesses. The use of corticosteroids had no influence upon mortality, but it was associated with a lower percentage of neurological sequelae (40% versus 14%; p < 0.05). The introduction of CT decreased mortality (40% in group A versus 23.5% in group B, although this difference was not significant) and also sequelae (86.6% in group A versus 57.6% in group B; p < 0.05). Leaving apart cases of bacterial endocarditis, in which death was due to the underlying heart disease and a systemic sepsis picture, mortality attributed to brain abscess was 20.3%. CONCLUSIONS: The introduction of CT has meant a significant breakthrough for the diagnosis, treatment and follow-up of these patients and has contributed to improvement in survival. In our series, the diagnosis of brain abscess was obtained earlier and the number of brain abscesses with no apparent source has decreased since the introduction of CT. Moreover, CT sensitivity is really good for locating multiple abscesses. Overall, the prognosis of these patients has improved since the introduction of this technique. Nevertheless, brain abscess is still associated with a relevant morbi-mortality rate. PMID- 9737148 TI - [Microbiologic diagnosis of Helicobacter pylori and its resistance to antibiotics]. AB - From March 1995 to February 1996 a total of 386 gastroduodenal biopsies were processed for microbiological diagnosis of Helicobacter pylori which included culture, Gram staining and urease test. For susceptibility studies to five antimicrobial agents, 35 additional gastroduodenal biopsies (n = 421) were added. There were 272 (70.4%) positive cultures, 220 (56.9%) samples with positive urase test and 244 (63.2%) with positive result in Gram-staining; both tests were statistically significant compared with culture (p < 0.05). Considering culture as the reference method, sensitivity and specificity values for the urease test were 77.0% and 92.1% and for Gram staining 86.7% and 92.9%, respectively. A total of 11 isolates were recovered from the 35 biopsies processed only for culture. Susceptibility testing of 283 isolates (272 + 11) was performed to the following antimicrobials: amoxicillin, metronidazole, clarithromycin, azythromycin and tetracycline. Resistance to metronidazole was 25.4% and the corresponding values for clarithromycin and azithromycin 9.5%. No resistance to amoxicillin or tetracycline was observed. Urease test and Gram staining are two easy-to-perform tests and when taken together allow the microbiological diagnosis of Helicobacter pylori infection. Culture should be performed to know the evolution of resistance to antimicrobials used for treatment of this infection. PMID- 9737149 TI - [Validation of the Spanish version of the diagnostic questionnaire of the Working Group on Atopic Dermatitis of the United Kingdom]. AB - BACKGROUND: Although there are several diagnostic questionnaires for atopic dermatitis (AD), they have not yet been validated or cannot be used in population studies. Our objective was to validate the questionnaire developed by United Kingdom Working Party's Diagnostic criteria for Atopic Dermatitis (UKWPDCAD). METHODS: Validation was developed in two steps: first, the questionnaire was adapted by means of translation and back-translation. Second, validation was conducted with 237 patients referred to the hospital with diagnoses unknown to the investigators. Patients were independently assessed by the investigators who were blinded to the questionnaire. RESULTS: A total of 102 cases of AD were diagnosed among the 237 patients. The questionnaire showed a sensitivity of 76.5%, with a 95% confidence interval (CI) of 66.8%-84.1%. The specificity was 90.4% (CI = 83.8%-94.6%); the positive predictive value was 85.7% (CI = 76.4% 91.8%), and the negative predictive value 83.6% (CI = 76.3%-89%). CONCLUSIONS: The spanish version of the questionnaire has shown a high diagnostic accuracy, similar to the original written in english. We consider it a useful tool to be used in frequency studies of AD in large general populations. PMID- 9737150 TI - [Estimating the costs of health care for patients infected with the human immunodeficiency virus in a university hospital in Catalonia]. AB - BACKGROUND: To estimate the use of resources and costs of health care to patients infected with the human immunodeficiency virus (HIV). METHODS: Prospective study in university hospital in Catalonia including 166 patients. AIDS was defined following 1987 criteria of the Centers for Disease Control. AIDS phase was divided into three grades according to the evolutive course: AIDS grade I, II and III. Resources/costs were calculated in function of the degree of disease, transmission mode and demographic variants. RESULTS: The mean cost per patient/year (PY) was 1,571,900 pesetas, ranging from 88,700 for the asymptomatic phase and 2,561,000 per AIDS phase. Within the AIDS phase costs ranged from 1,593,317 for AIDS grade I to 4,903,183 for grade III. This increase was due to differences in hospitalization days for PY (up to 12.4 days in pre-AIDS phases, 45.8 for AIDS phase I and 119.4 for AIDS phase III) and days in day-hospital per PY (38.1 days for AIDS grade III). Parenteral drug abusers (PDA) had a PY cost 42% lower than that corresponding to sexually infected patients. CONCLUSIONS: Health care costs per PY of HIV infected patients increase with disease progression, which relates to the increase in hospitalization days and day hospital hospitalization that occurs in the advanced phases of the disease. Our results suggest that health care to PDA is cheaper than that for sexually infected patients. PMID- 9737151 TI - [Soft tissue infections with S. Lugdunensis. Presentation of 2 cases and general review]. AB - Staphylococcus lugdunensis was first described in 1988 by Freney et al. Since that moment, it has been recognised as an important cause of endocarditis that is able to involve native valves and follow an aggressive, often fatal course. By now, we know only few potential virulence factors, but its ability to produce a wide range of human infections, varying from life threatening (such us endocarditis) to more benign ones, is unquestionable. Unlike other coagulase negative Staphylococci, S. lugdunensis is frequently implicated in soft tissue infections and affects commonly persons with clinical antecedents of neoplastic diseases. We report two new cases of deep tissue abscesses in patients that had history of uterine tumours. In both occasions we isolated coagulase negative Staphylococci that were markedly susceptible to all the antimicrobial agents tested. It is relevant to perform a detailed microbiological study in order to avoid misidentification of this species, since it differs significantly from other coagulase negative ones and may lead to important morbidity--and even mortality--. There are great differences in the rates of resistant strains between countries and, up-to-date, the lack of antibiotics susceptibility seems not to be a serious problem in Europe if we compare with the 30% american strains that are resistant via production of beta-lactamase. PMID- 9737152 TI - [Resistant Mycobacterium tuberculosis: its relation with coinfection with human immunodeficiency virus. 5-year study]. AB - BACKGROUND: To analyze the situation of resistance to antituberculous drugs among strains of Mycobacterium tuberculosis recovered in our environment during a five year period and its relationship with HIV co-infection. PATIENTS AND METHODS: Review of the Mycobacterium tuberculosis strains recovered from patients aged older than 14 years in Hospital Joan March from 1992 to 1996 of which a susceptibility testing study by means of the multiple proportions method was available. The initial or secondary resistance was considered according to the previous antituberculosis treatment antecedent. RESULTS: The susceptibility testing was available from 179 cases (136 males and 43 females) out of a total of 214. The overall resistance rate to any of the tested drugs was 10.1% (18 cases) with a 4% (6 cases) of initial and 38.7% (12 cases) secondary resistances. Co infection with HIV showed not to be a risk factor for the development of resistance. No significant increase was observed analyzing the temporal trend through the five years studied. CONCLUSIONS: At present, the situation of resistance to Mycobacterium tuberculosis seems not to be alarming in our environment. Co-infection with HIV has not been shown to be associated with an increase in resistance rates. PMID- 9737153 TI - [Usefulness of CO measurement in expired air in the study of tobacco consumption by youths and adolescents]. AB - The objective of our work was to know the relationship between carbon monoxide (CO) levels in expired air and smoking habits among school youths and the relationships that can be established between CO level and some peculiar attitudes regarding consume by youths, such as the number of cigarettes, inhaling technique and time elapsed since the last cigarette was smoked. The study, of cross-sectional design, was performed in two high school centres and a total of 777 students who answered a questionnaire and had an expired air CO sample in their own schoolroom tested were enrolled. CO determination in the schoolroom was a simple and attainable technique for the pupils, as only 32 cases (4.1%) had to be excluded due to poor collaboration or poor technique. The mean (mean and SD) CO level in the control group (n = 247), made up by non-smokers nor tobacco tasters was 4.75 (2.46) ppm, statistically lower than among smokers (p < 0.001), but with no differences compared with non smokers (n = 563), who had a CO level of 5.23 (3.4) ppm. This figure was also lower (p < 0.001) than that obtained in the smokers (12.6 [6.3] ppm), made up of 214 pupils, with a mean consume of 2.7 (1.69) cigarettes/day. Among smokers the mean abstinence time since the last cigarette was smoked was 26 (44) minutes and 54% of them admitted to have smoked in the last 10 minutes. CO in expired air correlated significantly with the number of smoked cigarettes (r = 0.58; p < 0.001). Likewise, it correlated significantly with abstinence minutes (r = -0.38; p < 0.001). The time required for CO level to decrease below 10 ppm was 140 minutes in four cases and 120 minutes in 33 cases. PMID- 9737154 TI - [Molecular epidemiology of the human immunodeficiency virus]. PMID- 9737155 TI - [Lichen planus of the oral mucosa]. PMID- 9737156 TI - [Current approach to the diagnosis and treatment of Helicobacter pylori infection in peptic ulcer disease]. PMID- 9737157 TI - [Pain in the inguinal region in a 24-year-old woman]. PMID- 9737158 TI - [A woman positive for the human immunodeficiency virus with a bilateral preauricular mass]. PMID- 9737159 TI - [Nodular cutaneous lesions in a patient with human immunodeficiency virus infection]. PMID- 9737160 TI - [Dementia and walking disorders in a middle-age patient]. PMID- 9737161 TI - [Paraplegia and meningitis in a patient with mediastinal and abdominal cystic mass]. PMID- 9737162 TI - [39-year-old male with fever and bilateral pleural effusion]. PMID- 9737163 TI - [Physiopathogenic justification of hematologic disorders in POEMS syndrome]. PMID- 9737164 TI - [Venous infarct as presenting form of venous angioma of the posterior fossa]. PMID- 9737165 TI - [Imaging diagnosis in Cushing's syndrome: a call to attention]. PMID- 9737166 TI - [Usefulness of fiber-bronchoscopy in the diagnosis of pulmonary hydatidosis]. PMID- 9737167 TI - [Esophageal bezoar: an exceptional complication of enteral nutrition]. PMID- 9737168 TI - [Addison's disease and hepatic biochemical disorders. Retrospective analysis]. PMID- 9737169 TI - [Hyperkinetic movements in ischemic ictus]. PMID- 9737171 TI - [Academic medicine--alternative medicine from the viewpoint of public health public health directors]. PMID- 9737172 TI - [Academic medicine and complementary medicine--campaign for the truth in medical theories]. PMID- 9737173 TI - [The academic medicine--alternative medicine tension field]. PMID- 9737174 TI - [Are academic medicine-alternative medicine incompatible, compatible, complementary?]. PMID- 9737175 TI - [Research in complementary medicine: results and problems]. PMID- 9737176 TI - [Subcutaneous hematomas, reduced Quick value. Factor VII deficiency]. PMID- 9737177 TI - [Mononucleosis infectiosa (Epstein-Barr virus infection]. PMID- 9737178 TI - New Medicare + Choice program brings challenges and opportunities. PMID- 9737179 TI - Obesity, a complicating factor in this diagnosis. PMID- 9737180 TI - Prostitution in Memphis--past, present, and future. PMID- 9737181 TI - Vanderbilt experience with cryosurgery for 25 advanced hepatic tumors. AB - There are reports that suggest cryosurgical techniques may be a useful adjunct or even a viable alternative to surgical resection for hepatobiliary malignancies. Our objective was to evaluate the clinical results following cryoablation in conjunction with surgical resection for advanced hepatic tumors. Cryosurgical techniques were used in 25 consecutive patients with advanced liver tumors (1) to achieve a > 1-cm tumor-free margin when standard surgical margins were close, (2) to manage multiple tumor nodules with or without standard surgical resection, or (3) to increase chemotherapy response rates in conjunction with hepatic arterial portocath placement. In these 25 patients cryoablation was applied to 44 of 91 lesions--independently in four patients and in combination with hepatic resection in 21 patients. Cryoablation was used in seven patients because of close surgical margins. In 18 patients cryosurgery was used for complete lesion ablation. In 14 of the 18 patients cryosurgery and resection were used for different lesions; in four cryosurgery alone was used. Transient changes in hepatic enzymes, PT, PTT, and platelets were at maximum on postoperative days 1 to 3. Surgical mortality and morbidity rates were 4% and 68% respectively. Coagulation abnormalities were common; at least 30% reduction in platelets occurred in all patients and a > 50% reduction occurred in 15 of 25 (60%). Sixteen patients had a PT > 15 sec and five of these 16 also had platelet count < 50,000. Associated complications included one wound hematoma, one GI hemorrhage, one intracranial hemorrhage, and one hepatic hemorrhage from the cryosurgical site. 96%, 66%, 49%, 35%, and 20% of patients were surviving respectively at 6, 12, 18, 24, and 36 months. This report helps define the risks and results of cryosurgical ablation in conjunction with surgical resection for very advanced hepatobiliary tumors. Management of lesions contiguous to major blood vessels can include the Pringle maneuver or total hepatic vascular isolation. Cryoablation can be applied carefully as a complement to resection to achieve total tumor ablation in selected otherwise unresectable patients. PMID- 9737182 TI - Gunshot wound to the face causing unilateral facial nerve injury. PMID- 9737183 TI - Prevention of disability from asthma in children and young adults. PMID- 9737184 TI - Abdominal pain while on coumadin. PMID- 9737185 TI - 43rd Scientific meeting of the Commonwealth Caribbean Medical Research Council. Ocho Rios, Jamaica. April 22-25, 1998. Abstracts. PMID- 9737186 TI - [The role of prolactin in immunological processes]. AB - Recently many investigations have been carried out which indicate possible link between the neuroendocrine and immunologic systems. The obtained information points out that some hormones not mentioned before can modify the immunological answer. More recently, the anterior pituitary hormone, prolactin, has been shown to have immune-stimulating properties through prolactin receptors, which are found an different cells. Among others prolactin receptors are present on B and T lymphocytes, monocytes and thomocyte epithelial cells. Immunological effects of this hormone were observed in vitro in investigations on animal experimental model and in humans. There are some therapeutic implications concerning patients with rheumatoid arthritis and with systemic lupus erythematosus and diagnostic implications, that determination of prolactin concentration in serum may appear to be useful marker of rejection in human heart transplantation. Although neuroendocrine regulations of immunological balance are complicated but by the regulation of prolactin secretion we can suspect the possibility of the control of autoimmunological diseases activity. PMID- 9737187 TI - [How does carotid artery stenosis increase?]. AB - In 25 patients with carotid artery stenosis equal or higher than 40% colour doppler examinations were performed 105 times. The aim of this study was estimate the dynamics of progress of carotid artery stenosis. The mean time of observation was 22.5 months. In 11 (44%) patients there was no progress of stenosis, in 6 (24%) a leap progress over 30% of arterial lumen, and in 8 (32%) patients the progress of carotid artery stenosis were gradual and slow. The leap progress of carotid artery stenosis was usually caused by eruption or dissection of atherosclerotic plaque located in carotid bifurcation and this situation poses especially high risk of stroke. PMID- 9737188 TI - [The evaluation of lymphocyte subpopulation by flow cytometry in healthy children]. AB - Seeing that amount of experiences concerning the cytometry of lymphocyte subpopulation in particular age ranges is very low we considered that undertaking this subject is useful. By the method of flow cytometry 49 children at the age from 2 months to 15 years were studied. According to the results of our studies we found, starting from the third year of life decrease of mean percentage values of cells CD19+, progressive in relation to the age, increase of mean percentage values of cells CD2+, increase of percentage values of cells CD8+ and cells CD56++ and the decrease of mean values of the indicator Th/Ts. Remaining CD3, CD4 did not show any statistically significant differences in investigated age ranges. Determined by the method of flow cytometry the percentage and complete values of lymphocyte subpopulations in peripheral blood in investigated age ranges can make the reference values in the assessment of immunological state. PMID- 9737189 TI - [The evaluation of a threat of osteoporosis among the employees at the pharmaceutical firm "Jelfa" in Jelenia Gora]. AB - Nine hundred eight workers of Pharmaceutical Firm "Jelfa" at Jelenia Gora at the age over 35 were examined. Examinations were done in the form of inquiry, what gave all necessary data about age, weight, feeding habits, kind of work, fractures in the past and menstruations in women. The information were elaborated together with densitometric tests. We measured bone density (BMD) with densimeter DTX-200 (Osteometer) in the distal part of not dominant antebrachium. 23% of women and 37% of men had the results of bone density below the norm. PMID- 9737190 TI - [Ambulatory diagnosis of urinary incontinence among women: the role of a one-hour pad weigh test]. AB - The aim of our studies was the evaluation of diagnostic usefulness of the 1-hour pad weight test in the different types of female incontinence. We examined 189 women at the age from 16 to 74 with the incontinence in anamnesis. The test was performed accordingly to the I.C.S. guidelines. The positive result of the test (more that 2 grams increase in the pad weigh) was found in 136 (71.6%) of examined women; including moderate urine leak in 24%, big in 55.1% and severe in 20.6% of them. The biggest average increase in pad weigh was found in urinary incontinence associated with the urgency, corresponding to 88.9 g in mixed incontinence and to 67.4 g in genuine urge incontinence. The sensitivity, specificity and efficacy of the test was 78.6%, 72% and 77% respectively. The shortened 1-hour pad weigh test is a simple and effective examination, especially in the stress incontinence. It is less effective in genuine urge incontinence and giggle incontinence. The negative effect of the 1-hour test, when the patients complaints coexist, should be the indication to repeat the test or to perform the 24-hour test. In the doubtful cases, before the surgery, the diagnostics should be extended to the full urodynamic examination. PMID- 9737191 TI - [The effect of progressive incremental exercise on some parameters of hemostasis]. AB - In 18 sportsmen the platelet count, thrombin time, prothrombin time, activated partial thromboplastin time (APTT), cloting factors (F):I, VII, VIII and fibrin/fibrinogen degradation products (FDP) were measured before, immediately after progressive incremental exercise--and 30 min later. All post-exercise changes of the values were corrected for the plasma volume changes, which were calculated from hematocrit values. Platelet count, thrombin time, prothrombin time FI and FVII did not change significantly after exercise. Immediately post exercise APTT was significantly shortened, but FVIII and FDP were significantly elevated. 30 min later FDP level normalized but FVIII and APTT changes persisted. The obtained data show that the applied physical exercise caused shift of the existing balance between coagulation and fibrinolysis towards hypercoagulability. They also confirm the necessity of taking into account post-exercise hemostasis changes in cardiological rehabilitation and endurance training in adults. PMID- 9737192 TI - [Pediatric differences in the activity of endothelin 1 in physiology and pathology]. AB - In the paper authors gathered hitherto existing opinions concerning endothelin, its structure, biological role and mechanisms of action. A special attention was paid to children and the activity of endothelin in physiology and pathology in various stages of development. PMID- 9737193 TI - [Liver and endocrine system. Part I: pituitary-thyroid axis activity disorders]. AB - Recent studies have demonstrated that more and more young people suffer from liver diseases. There is a close relationship between endocrine system and the liver, where the hormone inactivation and synthesis of protein binding hormones takes place. Impairment of hepatocyte function may lead to disturbed homeostasis of the endocrine system. In part I current opinions on thyreometabolic state as well as plasma levels of pituitary-thyroidal axis hormones and their binding proteins in chronic hepatic disorders are presented. PMID- 9737194 TI - [Circulatory system in obese patients]. AB - The epidemiology of obesity and a variety of coexisting organism's functional disturbances was presented. Dynamic changes in circulatory system in obese patients, a development of hypertension as well as changes in morphology of the heart muscle itself and its function were described. Hyperinsulinism and coagulation disorders were considered as more important etiopathological factors leading to heart failure or hypertension rather than obesity alone. PMID- 9737195 TI - [Carpal tunnel syndrome: etiology, pathogenesis, diagnosis and treatment]. AB - The authors have showed a short nosography about carpal tunnel syndrome. History of research and treatment, pathogenesis, basic subjective and objective symptoms enabling diagnosis have been presented. Electromyography is a very important investigation to make a proper diagnosis. In differentiation between other diseases radicular syndromes of upper limb are very important, as they give similar symptoms and often are reasons of incorrect diagnosis and inappropriate treatment. PMID- 9737196 TI - [Vagal nerve neurinoma: a case report]. AB - A case of asymptomatic 44-year-old male patient with moderate arterial hypertension, misdiagnosed as aortic aneurysm, is reported. Additional diagnostics (transthoracic and transesophageal echocardiography, CT scan) revealed the presence of mediastinal tumor, diagnosed at operation as vagal neurinoma. Early and late results of therapy are described. PMID- 9737197 TI - [Buttock claudication as the dominant symptom of atherosclerosis: a case report]. AB - Buttock claudication is rather rare symptom of atherosclerosis. Authors presented a case of 61 year old patient in whom buttock claudication was only one symptom. The patient was surgically treated (endarterectomy of iliac internal artery with ilico-femoral bypass) with excellent result. Authors pointed out necessity of restoration of blood flow through internal iliac artery as a prevention or treatment in buttock claudication. PMID- 9737198 TI - [Five cases of acquired pure red cell aplasia]. AB - Acquired pure red cell aplasia is rare disorder of hematopoiesis. Because of the variety of reasons for PRCA optimal treatment is based on highly specialistic diagnostics. Both humoral and cellular mechanisms are involved in the pathogenesis of PRCA. Corticosteroids, cyclosporin A, high-dose intravenous immunoglobulin therapy are recommended in PRCA. Treatment failures may be successfully managed with antihuman thymocyte globulin, cyclophosphamide or azathioprine. We describe also the lots of five patients with this disorder. PMID- 9737199 TI - [Andrew Victor Schally: neuroendocrinologist of Polish origin]. AB - American scientist of Polish origin, won the Nobel Prize in the field of physiology and medicine. In this way he became one of three Polish Nobel Prize winners in the scientific field (among M. Sklodowska-Curie and Tadeusz Reichstein). On the 20th anniversary of this historic moment I describe his life, scientific activity and achievements. PMID- 9737200 TI - Applications of reporter genes. PMID- 9737201 TI - Color images for fast-scan CV measurements in biological systems. PMID- 9737202 TI - Getting under the skin: implantable glucose sensors. PMID- 9737204 TI - An electrothermal atomic absorption spectrometer using semiconductor diode lasers and a tungsten coil atomizer: design and first applications. AB - A new type of atomic absorption spectrometer using a laser diode as light source and a tungsten coil as atomizer is described. Compared to established atomic absorption spectrometers, it is much simpler in construction, smaller in size, and less expensive and it provides inherent background correction and high detection power. The performance of this concept is demonstrated by the determination of aluminum and chromium in water, blood serum and, using the slurry sampling technique, in powdered high-purity graphite and titanium dioxide samples. For calibration, the standard addition method was used. Possible interferences by impurities originating from the tungsten coils are discussed. Applying aqueous solutions of Al and Cr, detection limits of 0.9 and 0.03 ng/mL, respectively, were obtained, and for serum, they were 2.5 and 0.3 ng/mL, respectively. For these elements in graphite and titanium dioxide applied as slurry, the detection limits are between 0.02 (Cr in TiO2) and 0.6 micrograms/g (Al in graphite). The accuracy was checked by comparison of the results with those of other methods. The described system is especially suitable for on-site and on-line analysis. PMID- 9737203 TI - Separation and characterization of amines from individual atrial gland vesicles of Aplysia californica. AB - Several amine-containing components of individual vesicles from the atrial gland of Aplysia californica were identified with capillary electrophoresis (CE). On line derivatization with naphthalene-2,3-dicarboxaldehyde was performed, and the derivatized amine-containing components were detected with laser-induced fluorescence (LIF). Amino acids, including taurine, that had not been determined previously in atrial gland vesicles were observed by using CE-LIF, and their identities were confirmed with CE, HPLC, NMR, and electrospray ionization mass spectrometry. The finding that taurine is packaged and stored into secretory vesicles supports the hypothesis that taurine may exhibit neuromodulatory activity. The bioactive peptides, well-known to be in atrial gland vesicles, were detected in lysed vesicle samples fractionated with HPLC and analyzed by matrix assisted laser desorption/ionization time-of-flight mass spectrometry. These peptides were also observed in single-vesicle runs with CE-LIF. The atrial gland vesicles (ranging from 0.5 to 2 microns diameter and 65 aL to 4 fL volume, respectively) studied in this work represent the smallest biological entities to be analyzed chemically on an individual basis. PMID- 9737205 TI - Simplification of product ion spectra derived from multiply charged parent ions via ion/ion chemistry. AB - High-mass multiply charged ions fragment to yield a mixture of products of varying mass and charge. When the measurement of mass-to-charge ratio is used to determine products ion mass, product ion charge must first be established. To minimize charge-state ambiguity in product ion spectra derived from multiply charged parent ions, product ions have been subjected to proton-transfer reactions with oppositely charged ions to reduce product ion charge states largely to +1. This procedure greatly simplifies the interpretation of product ion spectra derived from multiply charged ions. Illustrative data are presented for the +4 and +3 parent ions derived from electrospray of melittin and the +12 to +4 parent ions of bovine ubiquitin, whereby product ions were formed in a conventional quadrupole ion trap tandem mass spectrometry experiment. Data are also shown for product ion mixtures derived from interface-induced dissociation of multiply charged ions derived from bovine ubiquitin, tuna cytochrome c, bovine cytochrome c, and equine cytochrome c. The use of ion/ion chemistry to simplify product ion spectra derived from multiply charged parent ions significantly extends the size range of macromolecules for which the quadrupole ion trap can derive structural information. PMID- 9737206 TI - A microfabricated dialysis device for sample cleanup in electrospray ionization mass spectrometry. AB - A laser microfabricated device was constructed for rapid microdialysis cleanup of biological samples for analysis by electrospray ionization mass spectrometry (ESI MS) in both off-line and on-line modes. A microdialysis membrane was sandwiched between two chips having micromachined serpentine channels. The total volume of the sample serpentine channel used for the microdialysis was 1 microL. Efficient desalting was demonstrated for both DNA and protein samples using ESI with an ion trap mass spectrometer after microdialysis against a counter flow of ESI compatible buffer. Signal-to-noise ratios were also greatly enhanced compared to direct infusion of the original nondialyzed samples. Importantly, the microfabricated device allowed use of sample flow rates 1 order of magnitude smaller than previous designs based on a microdialysis fiber, allowing reduced sample utilization and improved sensitivity with ESI-MS. The effectiveness of the cleanup was attributed to the size difference between the sample channel and the buffer channel and the fact that the sample is continuously refreshed by the buffer counterflow. The results indicate substantial potential for construction of highly compact and rugged devices enabling field applications of ESI-MS. PMID- 9737207 TI - Method to compare collision-induced dissociation spectra of peptides: potential for library searching and subtractive analysis. AB - We report the development of a method to compare collision-induced dissociation (CID) spectra of peptides. This method employs a cross-correlation analysis of a CID spectrum to a reference spectrum and normalizes the cross-correlation score to the autocorrelation of the CID spectra. The query spectrum is compared by using both mass information and fragmentation patterns. Fragmentation patterns are compared to each other using a correlation function. To evaluate the specificity of the approach, a set of 2180 tandem mass spectra obtained from both triple-quadrupole tandem mass spectrometers (TSQ) and quadrupole ion trap mass spectrometers (LCQ) was created. Comparisons are performed between tandem mass spectra obtained on the same instrument type as well as between different instrument types. Accurate and reliable comparisons are demonstrated in both types of analyses. The scores obtained in the cross-comparison of TSQ and LCQ tandem mass spectra of the same peptide are found to be slightly lower than comparisons performed with spectra obtained on the same instrument type. The method appears insensitive to variations in day-to-day performance of the instrument, minor variations in fragment ion abundance, and instrumental differences inherent in the same instrument model. The use of this method of comparison is demonstrated for library searching and subtractive analysis of tandem mass spectra obtained during LC/MS/MS experiments. PMID- 9737208 TI - Hydrogen/deuterium exchange of nucleotides in the gas phase. AB - Gas-phase hydrogen/deuterium exchange reactions have been performed on the 5'- and 3'-nucleotide monophosphates and on the 3'5'-cyclic nucleotides. Following negative mode electrospray ionization and transport to a Fourier transform ion cyclotron resonance cell, each nucleotide was reacted with gaseous D2O for up to 600 s. Extensive deuterium exchange was observed for the 3'- and 5'-nucleotides in negative ion mass spectra with relative rates of exchange following the trend 5'dCMP > 5'-dAMP > 5'dTMP >> 5'-dGMP and 3'-dGMP > 3'-dAMP approximately equal to 3'-dCMP approximately equal to 3'-dTMP. At least two classes of exchanging protons are observed. The more facile class is assigned to the amino protons of the bases, with a slower class attributed to the phosphate and/or hydroxyl proton. Overall, the 3'-nucleotides exchange more quickly than the 5' oligonucleotides. The cyclic nucleotides did not undergo deuterium exchange, suggesting that a charged phosphate group proximate to the base is required to catalyze the exchange reaction. Exchange through tautomerization of the bases is no observed, although molecular modeling suggests an energy barrier of < 30 kcal. PMID- 9737209 TI - Chemiluminescence flow-through sensor for copper based on an anodic stripping voltammetric flow cell and an ion-exchange column with immobilized reagents. AB - A novel chemiluminescence (CL) flow-through sensor based on immobilizing all the ingredients involved in the analytical reaction for the determination of copper is proposed. The analytical reagents including luminol and cyanide were coimmobilized permanently on an anion-exchange column, while the analyte copper was retained temporarily by electrochemical preconcentration on a gold electrode placed in an anodic stripping voltammetric flow cell. By injection of a volume of sodium hydroxide through the column with immobilized reagents, luminol and cyanide were eluted from the resins in alkaline aqueous solution and then reacted with copper stripped from the gold electrode to produce a CL signal, by means of which copper could be sensed. The sensor was not susceptible to interference by other metal ions associated with the CL reaction. The response to the concentration of copper was linear in the range of 0.01-10 micrograms/L and an extremely low detection limit of 8.0 x 10(-3) micrograms/L was achieved. A complete analysis could be performed in 4 min with a relative standard deviation of less than 8%. The column with immobilized reagents was readily prepared and could be reused over 200 times. The sensor was applied successfully to the determination of copper in human serum and natural water samples. PMID- 9737210 TI - Quantitative analysis of chemical warfare agent degradation products in reaction masses using capillary electrophoresis. AB - Quantitative methods have been developed for the analysis of chemical warfare agent degradation products in reaction masses using capillary electrophoresis (CE). This is the first report of a systematic validation of a CE-based method for the analysis of chemical warfare agent degradation products in agent neutralization matrixes (reaction masses). After neutralization with monoethanolamine/water, the nerve agent GB (isopropyl methylphosphonofluoridate, Sarin) gives isopropyl methylphosphonic acid (IMPA) and O-isopropyl O'-(2 amino)ethyl methylphosphonate (GB-MEA adduct). The nerve agent GD (pinacolyl methylphosphonofluoridate, Soman), [pinacolyl = 2-(3,3-dimethyl)butyl] produces pinacolyl methylphosphonic acid (PMPA) and O-pinacolyl O'-(2-amino)ethyl methylphosphonate (GD-MEA adduct). The samples were prepared by dilution of the reaction masses with deionized water before analysis by CE/indirect UV detection or CE/conductivity detection. Migration time precision was less than 4.0% RSD for IMPA and 5.0 RSD for PMPA on a day-to-day basis. The detection limit for both IMPA and PMPA is 100 micrograms/L; the quantitation limit for both is 500 micrograms/L. For calibration standards, IMPA and PMPA gave a linear response (R2 = 0.9999) over the range 0.5-100 micrograms/mL. The interday precision RSDs were 1.9, 1.0, and 0.7% for IMPA at 7.5, 37.5 and 75.0 micrograms/mL, respectively. Corresponding values for PMPA (again, RSD) were 2.9, 1.1, and 1.0% at 7.5, 37.5 and 87.5 micrograms/mL, respectively, as before. Analysis accuracy was assessed by spiking actual neutralization samples with IMPA or PMPA. For IMPA, the seven spike levels used ranged from 20 to 220% of the IMPA background level, and the incremental change in the found IMPA level ranged from 86 to 99 % of the true spiking increment (R2 = 0.9987 for the linear regression). For PMPA, the five spike levels ranged from 10 to 150% of the matrix background level, and similarly, the accuracy obtained ranged from 95 to 97% of the true incremental value (R2 = 0.9999 for the linear regression). PMID- 9737211 TI - Base stacking: pH-mediated on-column sample concentration for capillary DNA sequencing. AB - An on-column sample concentration method for capillary-based DNA sequencing was studied. This base-stacking method allows direct injection of unpurified products of dye-primer sequencing reactions onto capillaries without any pretreatment. On column concentration of DNA fragments is achieved simply by electrokinetic injection of hydroxide ions. A neutralization reaction between these OH- ions and the cationic buffer component Tris+ results in a zone of lower conductivity, within which field focusing occurs. DNA fragments are concentrated at the front of this low-conductivity zone. With sample injection times as long as 360 s at 50 V/cm, resolution could still be restored by the stacking process. Using a 36/47 cm-long uncoated capillary, with poly(dimethylacrylamide) as the separation matrix, and electric field of 160 V/cm, a resolution of at least 0.5 could be generated for fragments up to 650 nucleotides long. Both resolution and signal strength are excellent relative to conventional injection of highly purified samples. No significant degradation of the capillary performance was observed over at least 20 sequencing runs using this new sample injection methods. PMID- 9737213 TI - Polishable and renewable DNA hybridization biosensors. AB - Routine applications of DNA hybridization biosensors are often restricted by the need for regenerating the single-stranded (ss) probe for subsequent reuse. This note reports on a viable alternative to prolonged thermal or chemical regeneration schemes through the mechanical polishing of oligonucleotide-bulk modified carbon composite electrodes. The surface of these biocomposite hybridization biosensors can be renewed rapidly and reproducibly by a simple extrusion/polishing protocol. The immobilized probe retains its hybridization activity on confinement in the interior of the carbon paste matrix, with the use of fresh surfaces erasing memory effects and restoring the original target response, to allow numerous hybridization/measurement cycles. We expect that such reusable nucleic acid modified composite electrodes can be designed for a wide variety of biosensing applications. PMID- 9737212 TI - Comparison of the retention characteristics of aromatic and aliphatic reversed phases for HPLC using linear solvation energy relationships. AB - The similarities and differences in retention characteristics of aromatic and aliphatic phases have been elucidated by the use of linear solvation energy relationships (LSERs). Three aromatic phases and three aliphatic phases were investigated in a series of mobile phases. The results of LSERs on a polymer based aromatic phase, poly(styrene-divinylbenzene) resin (PRP-1) are very different from those on either silica- or zirconia-based aromatic and aliphatic phases. Retention on all aromatic and aliphatic phases except PRP-1 is dominated by two factors: the solute size and hydrogen bond acceptor basicity. On the other hand, in addition to these two major contributions, retention on PRP-1 is markedly influenced by the solute hydrogen bond donor acidity. We believe that PRP-1 exhibits a more adsorption-like retention mechanism than do the other phases. With the inorganic oxide-based phases, the aromatic phases are less retentive than the aliphatic phases but show a larger dependence on molecular polarizability. The enhanced polarizability of aromatic phases is the likely cause of some differences in their chromatographic selectivity relative to the aliphatic phases. PMID- 9737214 TI - Bubble fractionation of enantiomers from solution using molecularly imprinted polymers as collectors. AB - Adsorptive bubble separation methods have been used to enrich components from both heterogeneous and homogeneous solutions. These methods are particularly effective for processing large solution volumes at low cost. Previous work demonstrated that chiral, surface-active collectors could be used to enrich enantiomers from homogeneous solution in a foam fractionation process. In a significant extension of this work, the use of highly selective molecularly imprinted polymers (MIPs) and heterogeneous solutions for the bubble flotation of enantiomers was evaluated. The high selectivity and ease of recycling of the MIP make this a potentially powerful approach for process-scale separations from large-volume bulk solutions. New MIPs were produced with low swelling properties which allowed them to retain enantioselectivity after numerous recyclings. PMID- 9737215 TI - 100th anniversary of the birth of Dr. Charles W. Mayo. PMID- 9737217 TI - Predicting clinical discordance of bone mineral density. AB - OBJECTIVE: To identify clinical and lifestyle factors that may predict clinical discordance of bone mineral density (BMD) in otherwise healthy perimenopausal and postmenopausal women referred for bone densitometry. MATERIAL AND METHODS: Data from 304 white women referred for bone densitometry were retrospectively reviewed in order to determine predictors of BMD status at the lumbar spine, femoral neck, and distal radius. In addition, a cross-validation study in a further independent sample of 50 patients was undertaken. Dual-energy x-ray absorptiometry of all three sites was performed, and T-scores were determined with use of standard criteria established by the World Health Organization. Covariables including age, postmenopausal status, years since menopause, use of alcohol and cigarettes, family history of osteoporosis, exercise, height, weight, and body mass index were analyzed by canonical discriminant functions. RESULTS: Seventy-six patients (25%) had normal BMD at all three sites (group A); 55 patients (18%) had osteopenia or osteoporosis at all sites (group B); and 173 patients (57%) showed regional discordance of BMD (group C). Menopausal status, years since menopause, use of alcohol and cigarettes, exercise levels, and weight allowed distinct separation of these three groups by using the plot of one canonical discriminant function against the other. When tested, this method of assignment correctly predicted the regional BMD status in 38 of 50 women (76%) in the independent sample. CONCLUSION: Thus, the combination of certain clinical and lifestyle factors may be helpful in predicting variations in the clinical classification of BMD in an ambulatory healthy perimenopausal or postmenopausal woman. PMID- 9737216 TI - Implementing nurse-based systems to provide American Indian women with breast and cervical cancer screening. AB - OBJECTIVE: To describe the factors critical to implementation of a nurse-based system to increase access for American Indian women to breast and cervical cancer screening. MATERIAL AND METHODS: We report the experience of 103 nurses at 40 clinics who were trained to use the nurse-based screening system. In addition, the critical elements are discussed in the context of one particularly successful site. RESULTS: Fifteen factors were identified as critical to the implementation of a nurse-based cancer screening process once a nurse had been trained to perform clinical breast examinations and collect Papanicolaou (Pap) test specimens: knowledge of benefit, skills, organization, adequate return, perceived patient demand, perceived effectiveness, legitimacy, confidence, commitment, adequate resources, a data-driven iterative approach to program implementation, an objective measure of quality, leadership, the passage of time, and a focus on delivering the service to the patient. For example, in one site that was particularly successful, the nurses, administrators, and other key health-care professionals contributed their respective resources to implement the screening program. The program was also supported by the lay community, the state board of nursing, and the state health department breast and cervical cancer control program. During the 3-year study period, the 103 nurses performed screening tests on 2,483 women, and only 18 of the Pap test specimens were unsatisfactory. CONCLUSION: Nurse-based systems designed to collect high-quality Pap test specimens and perform detailed clinical breast examinations can be implemented if the factors that are critical to implementation are identified and addressed. PMID- 9737218 TI - Middle cerebral artery territory infarction and early brain swelling: progression and effect of age on outcome. AB - OBJECTIVE: To determine the clinical course and outcome in patients with a middle cerebral artery (MCA) occlusion and early computed tomographic (CT) scan findings of infarction, particularly relative to age of the patient. MATERIAL AND METHODS: The clinical and neuroimaging features of 42 consecutive patients with MCA occlusion and early CT signs of swelling (within 24 hours after ictus) were studied. CT scans were graded for displacement of the pineal gland and septum pellucidum as well as compression of the frontal horn of the ventricular system. Young adults, defined as younger than 45 years of age, were assessed separately. RESULTS: Overall mortality was 55% in this patient population at risk for further neurologic deterioration. Of the 42 patients, 33 had deterioration-an impaired level of consciousness ensued in 3, a diencephalic herniation syndrome developed in 19, and uncal herniation occurred in 11. Mortality was 70% in these patients with deterioration. Mortality was significantly lower in younger patients with deterioration in comparison with older patients (3 of 11 patients versus 20 of 22; P = 0.00018, Fisher's exact test). Factors predictive of deterioration and poor outcome were older age (more than 45 years) and the presence of hyperdense clot in the MCA on CT scan, in addition to early swelling. CONCLUSION: Deterioration from further brain swelling is common in patients with MCA occlusion and sulci effacement on early CT scan. The outcome is fatal in most patients who deteriorate. Mortality was significantly higher in deteriorating older patients than in younger patients. Clearly defined criteria for decompressive hemicraniotomy in young patients with complete MCA occlusion are needed, preferably derived from a randomized clinical trial. PMID- 9737219 TI - Effect of thyroid hormone replacement on lipoprotein(a), lipids, and apolipoproteins in subjects with hypothyroidism. AB - OBJECTIVE: To study the effect of thyroid hormone replacement on total cholesterol, low-density lipoprotein cholesterol, triglycerides, high-density lipoprotein cholesterol (HDL-C), apolipoprotein A-I and B-100, and lipoprotein(a) [Lp(a)] in subjects with hypothyroidism. MATERIAL AND METHODS: In 17 patients with clinical primary hypothyroidism, studies were done before and after thyroid hormone replacement therapy. Free thyroxine and thyrotropin were determined by chemiluminescent assay. Total cholesterol and triglycerides were measured by enzymatic methods, and HDL-C was measured after dextran sulfate-MgCl2 precipitation. Apolipoprotein A-I and B-100 were assayed by immunonephelometry. For measurement of Lp(a), we used a sequential sandwich enzyme-linked immunosorbent assay. RESULTS: After levothyroxine treatment, the mean concentration of thyrotropin decreased from 91.4 to 3.7 microIU/mL, and free thyroxine increased from 0.5 to 1.2 ng/ dL. Total cholesterol, triglycerides, HDL C, low-density lipoprotein cholesterol, and apolipoprotein A-I and B-100 decreased after thyroid hormone replacement therapy. Lp(a) levels also decreased significantly (P<0.05) after treatment, from a mean of 33.4 to 25.6 mg/dL. CONCLUSION: Hypothyroidism is associated with an increase in total cholesterol, triglycerides, HDL-C, apolipoprotein A-I and B-100, and Lp(a). A reduction in lipid and lipoprotein levels after thyroid hormone replacement in our study cohort resulted in a less atherogenic profile. PMID- 9737220 TI - Osteosarcoma of the bones of the foot--an easily misdiagnosed malignant tumor. AB - OBJECTIVE: To evaluate the frequency of osteosarcoma involving the foot and determine the clinical outcome of affected patients. MATERIAL AND METHODS: We retrospectively reviewed the cases of osteosarcoma encountered between 1911 and 1992 at the Rizzoli Orthopedic Institute. In particular, we analyzed whether common clinical, radiographic, or histologic features could lead to a correct diagnosis of osteosarcoma of the foot. RESULTS: The bones of the foot were the primary site of osteosarcomas in 12 cases--0.6% of the entire series of such tumors at our institution during the study period. The mean age of the patients was 33 years. Initial symptoms were usually pain and swelling. Late diagnosis was common; the mean time interval between the first symptoms and diagnosis was 28 months. Misdiagnoses occurred in 6 of the 12 cases (50%): osteoblastoma, chondroblastoma, chondrosarcoma, osteoid osteoma, desmoid fibroma, and osteomyelitis were, respectively, the initial diagnoses. Histologically, 5 of the 12 tumors (42%) were low-grade lesions. Four of the seven patients with a high grade tumor died of metastatic disease after a mean survival of 50 months. Eight patients are alive with no evidence of disease after a mean follow-up of 162 months. CONCLUSION: When a painful swelling in a bone of the foot is observed, even if numerous benign conditions (such as fractures, infections, or benign bone tumors) are far more likely to occur, osteosarcoma must be ruled out to avoid delays in the treatment. Osteosarcomas of the foot may easily be misdiagnosed, especially because they almost always occur in adults, in contrast to osteosarcomas in general. High-grade tumors of the foot are as aggressive as other osteosarcomas and should be managed accordingly--with use of a safe-margins surgical procedure and chemotherapy. PMID- 9737221 TI - Randomized prospective pilot study of long-term dual-site atrial pacing for prevention of atrial fibrillation. AB - OBJECTIVE: To determine whether dual-site atrial pacing is feasible, safe, and effective. DESIGN: We undertook a randomized prospective single-blind crossover study. MATERIAL AND METHODS: Nine patients with at least two episodes per month of symptomatic paroxysmal atrial fibrillation participated in a randomized crossover study involving three separate 3-month blocks of single-site atrial pacing, dual-site atrial pacing, and control (support-only) pacing. RESULTS: Dual site atrial pacing resulted in shorter P wave duration (81 +/- 14 ms) than did single-site pacing (111 +/- 12 ms) or control sinus rhythm (123 +/- 9 ms) (P<0.0001) and in fewer premature atrial complexes on Holter monitoring (P = 0.06). The arrhythmia-free interval was longer with dual-site pacing (67 +/- 17 days) than with single-site (62 +/- 30 days) or support-only (49 +/- 34 days) pacing (P = 0.10). This pilot study was not statistically powered to detect a difference between pacing modes. CONCLUSION: (1) Dual-site atrial pacing is feasible and safe; (2) it shortens the P wave duration and tends to decrease premature atrial complexes on Holter monitoring; (3) any atrial pacing tends to prolong the arrhythmia-free interval; and (4) this pilot study enrolled too few patients to determine whether a significant difference in pacing modes exists and supports the need for a larger study. PMID- 9737222 TI - Developments in the treatment of alcoholism. AB - The treatment of alcoholism has changed during the past 2 decades. Notable developments have occurred in pharmacotherapy, psychotherapy, and health-care delivery. A better understanding of the biologic basis for addiction has led to clinical trials of medications that target neuroreceptors. One such medication is the opiate antagonist naltrexone, which decreases the craving for alcohol. Psychosocial interventions continue to be the mainstay of alcohol treatment programs. The efficacy of three different therapies was demonstrated in a study called Project MATCH (Matching Alcoholism Treatments to Client Heterogeneity). This study, however, did not prove the patient-treatment "matching" hypothesis. In addition to therapies provided by addiction specialists, interest is growing in the use of brief motivational techniques in primary-care settings. As the field of addiction responds to an unfolding health-care delivery system, a broader range of treatment options in conjunction with a greater opportunity to individualize patient care is evolving. PMID- 9737223 TI - Edgar Adrian--Nobel Prize for work on neurons. PMID- 9737224 TI - Primary immunodeficiencies. AB - The primary immunodeficiencies are congenital disorders that affect the function of the immune system. The result is an inadequate immune response to microorganisms, self-antigens, and tumor cells, which leads to increased susceptibility to infections, autoimmunity, or malignant disease. A substantial advance has been made in the understanding of the exact molecular mechanisms leading to primary immunodeficiencies; however, for some types, a specific genetic defect has not yet been determined. The life expectancy of patients with primary immunodeficiencies has increased considerably because of bone marrow transplantation and replacement therapies. Gene therapy has already been used for a particular type of immunodeficiency and is a promising alternative for the future management of many other types of primary immunodeficiencies. A better understanding of the genetic defects that lead to primary immunodeficiencies would result in the development of novel therapeutic strategies. PMID- 9737225 TI - Clinical recognition of pulmonary embolism: problem of unrecognized and asymptomatic cases. AB - Dyspnea, pleuritic chest pain, and tachypnea are widely appreciated as common initial features of pulmonary embolism (PE). This knowledge is derived primarily from prospective studies evaluating diagnostic tests or therapeutic interventions in which the study patients are suspected to have PE based on their initial symptoms. Autopsy studies, however, continue to show that most cases of fatal PE are unrecognized and undiagnosed. Data from studies screening for PE in patients with deep venous thrombosis and in postoperative patients suggest that many patients with PE are asymptomatic and that PE is unrecognized. We believe that the current concepts regarding the initial clinical features of PE are too narrow and biased toward symptomatic cases. High clinical suspicion may be insufficient in recognizing PE. Herein we summarize the available data and explore the implications for clinical practice. PMID- 9737226 TI - Evaluation and treatment of constipation and fecal impaction in adults. AB - Constipation is a common complaint that physicians encounter. Understanding the patient's definition of constipation and focusing the history and physical examination provide clues to the underlying cause. Initially, an empiric treatment trial is recommended. For patients with warning symptoms or those in whom treatment fails, a limited diagnostic work-up is suggested. Tests of physiologic function are reserved for patients whose condition is refractory to therapy. Fecal impaction can be considered extreme constipation. The pathophysiologic features of fecal impaction are discussed, and recommendations are provided for treatment and prevention. PMID- 9737227 TI - High-output heart failure due to a renal arteriovenous fistula in a pregnant woman with suspected preeclampsia. AB - A 29-year-old nulliparous woman had development of hypertension, proteinuria, and congestive heart failure during the third trimester of her pregnancy. Her symptoms and cardiovascular changes were consistent with congestive heart failure and severe preeclampsia. The underlying pathophysiology was believed to be caused by the high-output state of pregnancy and by the increased peripheral vascular resistance of preeclampsia. The patient underwent an elective cesarean section, but her cardiovascular symptoms did not resolve. Soon after delivery, the patient was found to have an arteriovenous fistula of the right renal artery that caused the high-output cardiac state. Embolization and surgical removal of the arteriovenous fistula resulted in complete resolution of the patient's high output heart failure. All previously reported cases of renal arteriovenous fistulas and malformations that have occurred during pregnancy are reviewed. PMID- 9737228 TI - Disseminated coccidioidomycosis associated with hypercalcemia. AB - This article describes a patient in whom disseminated infection with coccidioidomycosis was associated with hypercalcemia. The patient had a low level of 1,25-dihydroxyvitamin D and a suppressed parathyroid hormone value, an indication that the hypercalcemia was not mediated by vitamin D or parathyroid hormone. The episode resolved readily with administration of pamidronate, an outcome suggesting that this is effective treatment of hypercalcemia of this origin. On follow-up of the patient while he was receiving antifungal therapy for Coccidioides immitis, calcium values remained normal. PMID- 9737229 TI - Porphyria cutanea tarda and human immunodeficiency virus: two cases associated with hepatitis C. AB - Hepatitis C virus (HCV) was recently recognized as a probable etiologic factor for porphyria cutanea tarda (PCT). A review of the literature revealed 40 cases of PCT associated with the human immunodeficiency virus (HIV). In most of these cases, hepatitis C status was unknown. We describe two patients with PCT who were coinfected with HIV and HCV and discuss the interaction of these two viruses. A diagnosis of PCT, especially in a young patient, should prompt investigation for underlying HIV and HCV infection. Porphyrin studies should be performed in any patient with HIV and photosensitivity. Clinicians should be aware of the infectious risk associated with the vesicles and erosions in these patients. PMID- 9737230 TI - 73-year-old man with hepatomegaly and weight loss. PMID- 9737231 TI - Interstitial laser prostatectomy. AB - In an attempt to minimize the need for hospitalization and the associated perioperative and postoperative morbidity, alternatives to transurethral resection of the prostate (the standard treatment modality for benign prostatic hyperplasia) have been sought. Various types of laser prostatectomy have been proposed. Interstitial laser coagulation is performed by placing laser-diffusing fibers directly in the hyperplastic prostatic adenoma. The goal is to achieve coagulation necrosis within the adenoma, which causes the prostatic lobes to regress and thereby relieves the bladder outlet obstruction. Either the transurethral cystoscopic approach or the perineal approach can be used for laser application. Numerous published studies have shown that this laser procedure safely and effectively decreases symptoms of prostatism, increases the urinary flow rate, and reduces the volume of the prostate. Because of substantial tissue edema after treatment, catheter drainage may be necessary for 7 to 21 days. Although retrograde ejaculation has occurred occasionally (affecting from 0 to 11.9% of patients in reported studies) and uncomplicated urinary tract infections are common after interstitial laser coagulation, no cases of impotence or sustained incontinence have been described. Because interstitial laser coagulation is not associated with blood loss or intravascular fluid shifts and, if necessary, can be performed with a combination of local anesthesia and intravenous sedation, even high-risk patients are candidates for this procedure. PMID- 9737232 TI - Transurethral enzyme injection--future management of benign prostatic hyperplasia. AB - Although benign prostatic hyperplasia, a common condition among elderly men, has been effectively treated with transurethral resection of the prostate, this surgical procedure is associated with many well-recognized risks and complications. Because of this potential morbidity and mortality, various alternative treatment strategies for benign prostatic hyperplasia have been proposed. The use of enzyme solubilization and ablation of prostatic tissue to alleviate urinary outlet obstruction has proved effective in dogs and warrants investigation in human trials. Transurethral enzyme injection of the prostate has the potential for being a treatment modality with minimal invasiveness, limited requirements for anesthesia, and minimal associated toxicity for the management of benign prostatic hyperplasia. PMID- 9737233 TI - Neurologic contributions of Bayard T. Horton. PMID- 9737234 TI - New model for cancer screening in American Indian women. PMID- 9737235 TI - Nomenclature problems of gastrointestinal epithelial neoplasia. PMID- 9737236 TI - Reticulum cell neoplasms of lymph nodes: a clinicopathologic study of 11 cases with recognition of a new subtype derived from fibroblastic reticular cells. AB - Lymph nodes contain nonlymphoid accessory cells including follicular dendritic cells (FDCs), interdigitating dendritic cells (IDCs) and fibroblastic reticular cells (FBRCs). Neoplasms derived from FDCs are uncommon, and those of IDC origin are even more rare. We report the clinicopathologic features of 11 reticulum cell neoplasms, including 2 of FBRC origin. There were seven male patients and four female patients ranging in age from 13 to 73 years. All cases involved lymph nodes (cervical or supraclavicular-6 cases), (abdominal--2 cases), epitrochlear (1 case); two had more than one site of involvement (cervical lymph node and mediastinum--1 case, cervical and abdominal lymph nodes--1 case). One case of FDC tumor had concomitant Castleman's disease, plasma cell variant. Each neoplasm showed similar histology with oval-to-spindle-shaped cells in a storiform or fascicular pattern. Based on immunophenotypic findings, the neoplasms were classified as FDC (five cases), IDC (two cases), FBRC (three cases), and reticulum cell neoplasm, not otherwise specified (one case). The FDC tumors showed immunoreactivity for CD21 or CD35, vimentin, and CD68. The IDC tumors showed strong positivity for S-100 protein and variable positivity for CD68 and CD1a. The cases derived from FBRCs were positive for vimentin, desmin, and smooth muscle actin. The neoplasm classified as reticulum cell neoplasm, not otherwise specified had similar morphologic features but showed only equivocal positivity for CD68 and vimentin. Follow-up was available for 9 of 11 (82%) cases with a mean of 3.5 years. Four of five patients with FDC tumors were alive with disease when last seen; the fifth is alive and well with no evidence of disease at 4-year follow-up. One patient with IDC tumor had a recurrence in a different nodal site. Two patients with FBRC tumor were disease free at follow-up of 2 years and 8 years, respectively. The patient with reticulum cell neoplasm, not otherwise specified, was alive and disease free 8 years after diagnosis. PMID- 9737237 TI - Neuroendocrine differentiation is a common feature of thymic carcinoma. AB - Immunohistochemical evidence of neuroendocrine differentiation in the form of reactivity for synaptophysin, neuron-specific enolase, and/or chromogranin was found in 11 of 19 (58%) thymic carcinomas having the typical morphologic features of that tumor type. Four of these 19 cases were studied ultrastructurally, and neuroendocrine-type cytoplasmic dense-core granules were found in two. In contrast, 84 thymomas were negative for these markers, except for a focal immunoreactivity for neuron-specific enolase in areas of medullary differentiation in half of the lymphocyte-rich tumors. The results of this study show that in the thymus, similar to most other organs, neuroendocrine differentiation is not limited to tumors with an identifiable neuroendocrine appearance in hematoxylin-eosin-stained slides, such as carcinoid tumor and small cell carcinoma, but rather that it represents a common event shared by the major types of malignant epithelial tumors of that organ. PMID- 9737238 TI - Atypical or worrisome features in cellular neurothekeoma: a study of 10 cases. AB - Cellular neurothekeoma is a recently characterized benign cutaneous neoplasm arising usually on the upper trunk or head and neck of children or young adults. Typical histology is that of a lobulated dermal tumor composed of spindle and epithelioid cells, arranged in fascicles and nests, lacking immunoreactivity for S-100 protein, but usually being NK1/C3 positive. We present 10 new cases of cellular neurothekeoma with atypical histologic features that have not been described previously and that suggested the possibility of malignancy. The age range of affected patients was 1 to 44 years (median, 20.5 years); sites included the head and neck (three cases), the upper limbs (two cases), the lower limbs (two cases), and the trunk (two cases). Atypical findings in individual cases included large size (up to 6 cm), deep penetration (extending into skeletal muscle or subcutaneous fat, or both), diffusely infiltrative borders, vascular invasion, high mitotic rate, and marked cytologic pleomorphism. Clinical follow up was available in 7 of 10 cases. Although the atypical features raised concern about the biologic potential of these lesions, preliminary follow-up (1-5 years) has shown no recurrence and suggests that complete surgical excision of these lesions is curative. These new data expand the morphologic spectrum of cellular neurothekeoma. PMID- 9737239 TI - Histology and cellular kinetics of prostatic atrophy. AB - This study addresses two issues regarding prostatic atrophy: (1) the histologic features of atrophy as seen on needle biopsy results and how they affect the diagnosis of atrophy and (2) the cellular kinetics of atrophy and what it suggests about the mechanism of atrophy. We reviewed hematoxylin and eosin sections for 103 prostate needle biopsy specimens with atrophy. Each biopsy specimen was classified as either simple atrophy (large atrophic glands without crowding: 53 cases) or postatrophic hyperplasia (PAH) (crowded focus of small atrophic acini: 50 cases). Cell proliferation in both the atrophic and benign glands was evaluated in 103 cases by immunohistochemistry using antibodies against MIB-1. The TdT-mediated dUTP-biotin nick-end labeling technique was performed on 61 cases to quantitate apoptosis in atrophic and benign glands. Thirty-two percent of cases showed chronic inflammation, 21% showed acute inflammation, 14% showed nucleoli, and 1% showed mitoses. In comparison to simple atrophy, PAH contained more frequent prominent nucleoli (p < 0.0001) and acute inflammation (p < 0.0001), yet not chronic inflammation. In a multivariate analysis, acute inflammation and PAH pattern influenced the presence of prominent nucleoli. Staining for MIB-1 was greater in atrophic (27.5 cells/1000 cells) than in benign glands (3.5 cells/1000 cells), greater in PAH than in simple atrophy (p = 0.0015), and greater with acute (p = 0.05) but not chronic inflammation. In a multivariate analysis, only the pattern of atrophy and not acute inflammation was found to influence MIB-1. The rate of apoptosis was negligible in both the benign and atrophic glands, did not vary with pattern of atrophy, and did not correlate with MIB-1. Despite the atrophic appearance, atrophic glands in PAH show more proliferative activity than benign, nonatrophic glands and show no evidence of active involution, justifying the term "postatrophic hyperplasia" for this pattern of atrophy. Prominent nucleoli are seen more frequently in postatrophic hyperplasia, even in the absence of acute inflammation. To avoid a potential erroneous diagnosis of cancer, a constellation of features suggestive of malignancy should be considered, rather than relying on prominent nucleoli as the sole criteria for the diagnosis of prostate cancer. PMID- 9737240 TI - Uterine tumors resembling ovarian sex-cord tumors have an immunophenotype consistent with true sex-cord differentiation. AB - Seven examples of Clement and Scully's type II uterine tumors resembling ovarian sex cord tumors were studied immunohistochemically with markers of sex cord, steroid cell differentiation (inhibin, O13, A103), or both, and were found to be immunoreactive for one or more of these markers in all cases. There was also constant immunoreactivity for vimentin and for hormone receptors and variable positivity for keratin, actin, and desmin. These findings provide strong evidence for the presence of true sex cord elements in these tumors, the derivation of which remains speculative. PMID- 9737241 TI - Atypical teratoid/rhabdoid tumor of the central nervous system: a highly malignant tumor of infancy and childhood frequently mistaken for medulloblastoma: a Pediatric Oncology Group study. AB - Fifty-five patients with atypical teratoid/rhabdoid tumors of the central nervous system were studied to define the clinical and pathologic features of this newly described neoplasm. The lesion occurred primarily in children younger than 2 (mean age at diagnosis, 17 months). The neoplasms were located in the posterior fossa (36 patients) and the supratentorial compartment (17 patients) or were multifocal in both compartments (2 patients) at presentation. Histologically, the tumors were composed of small cells and large, pale cells in a jumbled architectural arrangement. The small cell component resembled medulloblastoma and occasionally had cords of cells in a mucinous background, simulating chordoma. The cytoplasm of the larger cells was conspicuous with a somewhat "rhabdoid" appearance, although rhabdoid features were not always prominent. Epithelioid features in the form of poorly formed glands or Flexner-Wintersteiner rosettes were noted in a minority of lesions. The neoplasms showed striking polyphenotypic immunoreactivity, including that for vimentin, glial fibrillary acidic protein, epithelial membrane antigen, cytokeratins, synaptophysin, chromogranin, and smooth muscle actin. Using a probe for chromosome 22, seven of eight scorable cases showed a solitary signal by fluorescence in situ hybridization (FISH) consistent with monosomy 22. The eighth scorable case showed three signals by fluorescence in situ hybridization and had a translocation involving chromosome 22 reported by conventional cytogenetics. In contrast to patients with medulloblastoma, the neoplasm with which these lesions are often confused, the outcome of the patients was uniformly poor. The mean postoperative survival of patients with atypical teratoid/rhabdoid tumors was only 11 months. Local recurrence, seeding of the cerebrospinal fluid pathways, or both, were common terminal events. This study underscores the distinctive clinical, histopathologic, immunohistochemical, and cytogenetic character of this unusually aggressive tumor. PMID- 9737242 TI - Proliferation and cellular phenotype in lymphomatoid granulomatosis: implications of a higher proliferation index in B cells. AB - Pulmonary involvement by lymphomatoid granulomatosis (LYG) is characterized by nodules of a polymorphous lymphoreticular infiltrate with necrosis, angioinvasion, and variable numbers of large, atypical cells. Using combined immunohistochemistry, the authors compared the expression of a marker of proliferation (DNA topoisomerase IIalpha) between B cells, T cells, and histiocytes. Sixteen cases of LYG were stained by combined immunohistochemistry for DNA topoisomerase IIalpha and CD-20, CD-3, CD-68, and CD-57. A proliferation index was determined for B cells, T cells, histiocytes, and natural killer cells by dividing the number of cells with coexpression of DNA topoisomerase IIalpha and CD-20, CD-3, CD-68, or CD-57 by the total number of CD-20+, CD-3+, CD-68+, or CD-57+ cells, respectively. A significantly higher proliferation index was present in B cells compared to T cells, histiocytes, or natural killer cells (p < 0.002). The average proliferation index for B cells was 0.25+/-0.24 (range, 0.00 0.76), for T cells was 0.02+/-0.01 (range, 0.00-0.04), for histiocytes was 0.00+/ 0.01 (range, 0-0.02), and for natural killer cells was 0.00+/-0.00 (range, 0.0 0.02). The average proliferation index of CD-20+ cells was greater in grade III LYG (0.36) than in grade II LYG (0.17) or the single case of grade I LYG (0.00). The authors conclude that (1) there is a spectrum of B-cell proliferation in LYG that roughly correlates with histologic grade, (2) T cells, histiocytes, and natural killer cells do not proliferate but are recruited, and (3) the average B cell proliferation index in grade III LYG is similar to that observed in large cell non-Hodgkin's B-cell lymphomas. These observations provide a possible rationale for the use of chemotherapy for grade III LYG and observation or immunologic adjuvants for LYG with grade I or grade II histology. PMID- 9737243 TI - Adenovirus colitis in human immunodeficiency virus infection: an underdiagnosed entity. AB - Adenovirus infection of the gastrointestinal tract in human immunodeficiency virus (HIV)-infected patients is rarely reported, probably because of a lack of familiarity of most pathologists with diagnostic criteria during routine light microscopy and possible misidentification as cytomegalovirus infection. We studied colonoscopic biopsy specimens from 135 HIV-infected patients with clinically suspected cytomegalovirus colitis during a 4.5-year period to morphologically identify the presence of adenovirus infection. Immunohistochemical staining for adenovirus was performed for confirmation on all suspected cases. Adenovirus infected cells showed characteristic amphophilic or eosinophilic nuclear inclusions, predominantly affecting the surface epithelium and characteristically involving goblet cells. Sixteen cases showed morphologic features of adenovirus infection, all confirmed by immunohistochemistry. Twelve cases also showed cytomegalovirus infection, whereas 4 showed adenovirus alone. In 10 cases, adenovirus colitis was not recognized during initial routine histopathologic diagnostic evaluation. Adenovirus inclusions also were discovered in the stomach, the duodenum, and the liver in single cases. Conclusions are as follows: (1) Adenovirus colitis has been underdiagnosed at our institution and, we suspect, in general. (2) The morphologic features and nuclear inclusions of adenovirus colitis are characteristic and can be identified reliably by routine light microscopy. (3) Adenovirus infection also may be diagnosed morphologically in extracolonic sites, such as the stomach, the small intestine, and the liver. (4) Coinfection of adenovirus with cytomegalovirus and other agents is seen frequently, but, less frequently, adenovirus may be identified as a sole pathogen. PMID- 9737244 TI - Nephroblastoma-like tumors in patients with testicular germ cell tumors. AB - Nephroblastoma-like tumors (NLTs) developed in metastatic sites in nine men with testicular germ cell tumors (GCTs). These tumors had a characteristic "triphasic" admixture of primitive tubular structures, sometimes with a glomeruloid pattern, blastema and stroma. Skeletal muscle differentiation was apparent in two cases. Specific neuroendocrine markers (synaptophysin and chromogranin A) were negative. All patients were treated by surgical excision. Six patients were alive with no evidence of disease from 4 to 12 years after diagnosis of GCT and NLT. One patient was alive with disease 6 years after diagnosis of GCT and 3 years after diagnosis of NLT. One man who also had metastatic primitive neuroectodermal tumor (PNET) had short survival, and one patient died of postoperative infection. We conclude that patients with testicular GCTs in whom NLTs develop in metastatic sites often experience prolonged survival. Surgical excision appears to be adequate treatment for NLT arising in metastatic testicular GCT in most patients. It is important to distinguish NLTs from PNETs in metastatic GCTs because of the more aggressive course and the frequently fatal outcome of the latter. PMID- 9737245 TI - Immature teratomas in children: pathologic considerations: a report from the combined Pediatric Oncology Group/Children's Cancer Group. AB - Pediatric germ cell tumors (n = 135) with a major component of immature teratoma (IT) registered on Pediatric Oncology Group/Children's Cancer Group treatment protocols from 1990 to 1995 were reviewed. Sixty cases were pure IT with no malignant component and 75 were mixed tumors with a major component of IT. Foci of yolk sac tumor (YST) were present in all 75 mixed tumors; additional malignant components were present in 15. The IT component was as follows: 47% grade 3, 29% grade 2, 24% grade 1. There were no significant correlations between tumor grade and patient age by specific subsets or overall (all p > 0.10). Significant correlations were detected between stage and the presence of foci of YST (p = 0.0145) and grade and the presence of foci of YST (p < 0.001). Serum alpha fetoprotein concentrations were elevated at diagnosis in 96% of ovarian tumors with foci of YST and were mildly elevated (< 60 ng/dL) in only 16% of tumors without YST. Overall 2- to 6-year survival rate was 96% and was related to the presence of YST. Central pathologic review revealed aspects of morphologic diagnosis that were most frequently misinterpreted by contributing pathologists. These included the classification of differentiating tissues as immature and the failure to recognize two well-differentiated patterns of YST (the hepatoid pattern resembling fetal liver and the well-differentiated glandular pattern resembling fetal lung or intestine). Such foci were often overlooked. The authors conclude that the presence of microscopic foci of YST, rather than the grade of IT, per se, is the only valid predictor of recurrence in pediatric IT at any site. PMID- 9737246 TI - Primary mucinous carcinomas of the skin express TFF1, TFF3, estrogen receptor, and progesterone receptors. AB - Mucinous carcinoma may present at various sites, including the breast and the gastrointestinal tract. Rarely, such tumors arise within the skin. Comparatively, breast lesions are relatively common and usually associated with a good prognosis. When pure, they are typically estrogen (ER) and progesterone receptor (PR) positive and responsive to tamoxifen. The authors studied 12 mucinous carcinomas of the skin and compared the morphology with that of typical mammary lesions. The authors also evaluated for expression of estrogen receptor, progesterone receptor, and the mucus-associated peptides of the trefoil factor family (TFF), TFF1 (formerly pS2) and TFF2 (formerly SP), using immunohistochemistry. The localization of mRNAs for TFF1, TFF2, and TFF3 (formally ITF) was also studied in a subset of three tumors, using in-situ hybridization with S35 labeled riboprobes. The Grimelius stain was used to look for evidence of neuroendocrine differentiation. Eight resembled type A mucinous carcinomas of the breast, two resembled type B, and one had composite features. The 12th was a papillary neoplasm. The two type B tumors exhibited argyrophilia. All showed strong nuclear staining with the estrogen receptor antibody but a more varied pattern with antibodies to progesterone receptor and TFF1. None labeled for TFF2. The detection of TFF1 in mammalian skin is a novel finding. Cutaneous mucinous carcinoma shows strong similarities to its mammary counterpart, including expression of estrogen receptor, TFF1, and TFF3 mRNA. These observations suggest that some mucinous carcinomas of the skin might respond to antiestrogenic therapies. PMID- 9737248 TI - Mitochondrial neurogastrointestinal encephalomyopathy: diagnosis by rectal biopsy. AB - A 14-year-old girl with the mitochondrial neurogastrointestinal encephalopathy syndrome had an 8-year history of intestinal pseudoobstruction with abdominal pain, persistent vomiting, gastric and duodenal dilatation, and duodenal diverticulosis. The child appeared chronically malnourished and had severe growth failure. Multisystem involvement was evident with the presence of ptosis, external ophthalmoplegia, muscle wasting, peripheral neuropathy, and diffuse white matter disease seen on magnetic resonance imaging. Lactic acidosis and increased cerebrospinal fluid protein were observed. Mitochondrial enzyme analysis of fresh-frozen skeletal muscle revealed a respiratory chain defect. Molecular genetic studies showed multiple mitochondrial DNA deletions. Pathologic findings in the intestine included atrophy of the external layer of the muscularis propria and an increased number of abnormal-appearing mitochondria in ganglion and smooth-muscle cells. Microvesicular steatosis was observed in liver, skeletal, and gastrointestinal smooth muscle, and Schwann cells of peripheral nerve. Brightly eosinophilic inclusions in the cytoplasm of gastrointestinal ganglion cells were visible by light microscopy, which were confirmed to be megamitochondria by ultrastructural studies. This is the first report of abnormal mitochondria observed in intestinal ganglion and smooth-muscle cells in this syndrome. PMID- 9737247 TI - Diagnostic value of immunostaining for tryptase in patients with mastocytosis. AB - The term "mastocytosis" is used to describe a heterogeneous group of disorders characterized by abnormal growth and accumulation of mast cells (MCs). Cutaneous and systemic variants exist. Systemic mastocytosis may show an indolent or malignant clinical course. In malignant mastocytosis (MM), the diagnosis often is missed because the MCs are morphologically abnormal and lack metachromatic granules or the underlying histologic picture is complex. The cytoplasmic serine protease tryptase is produced by MCs and is thought to be expressed at all stages of MC maturation. To assess the diagnostic value of tryptase staining in mastocytosis, tissue sections from 93 patients with mastocytosis, including MM (n = 37), systemic indolent mastocytosis (n = 47), urticaria pigmentosa (n = 5), MC leukemia (n = 2), and solitary skin mastocytoma (n = 2) were stained with the antitryptase antibody G3. The results were compared with those of Giemsa and chloroacetate esterase (CAE) staining. Using antitryptase antibody G3, MC infiltrates were identified in all patients examined, including those with MM (37 of 37), and virtually all the neoplastic MCs (> 95%) appeared to react with G3. In MM, significantly fewer MCs were positive in Giemsa (54.5%; p < 0.05) and CAE (78.8%; p < 0.05). Moreover, G3 produced clear diagnostic staining in all cases of MM, but the proportion of cases with clear diagnostic results (> 10% of neoplastic cells positive) was considerably lower with Giemsa (48.6%; p < 0.05) and CAE (75.7%; p < 0.05) staining. By contrast, tryptase, Giemsa, and CAE produced diagnostic staining of MCs in virtually all cases of systemic indolent mastocytosis, urticaria pigmentosa, and solitary skin mastocytoma. In systemic mastocytosis, survival was significantly reduced in cases with Giemsa-/tryptase+ or CAE-/tryptase+ tumor cells compared to those cases with Giemsa+ or CAE+ MC infiltrates (p < 0.001). PMID- 9737249 TI - Pulmonary immunocytoma with massive crystal storing histiocytosis: a case report with review of literature. AB - An unusual immunocytoma (lymphoplasmacytoid lymphoma) composed predominantly of sheets of globoid and spindle-shaped crystal-storing histiocytes was detected incidentally in the right lung of a 72-year-old woman. Scattered lymphoplasmacytic aggregates within the tumor had monoclonality with anti-kappa immunoglobulin (Ig) M antibodies. The crystals were outlined positively by the same antibodies. They stained an intense blue with phosphotungstic acid hematoxylin (PTAH) and were found during electron microscopy to be membrane bound and also within type I pneumocytes and the extracellular space. Excessive production of kappa IgM by neoplastic low-grade lymphoplasmacytoid cells of B cell origin in an altered intra- or extracellular milieu may lead to crystallization and phagocytosis by reactive histiocytes. Review of the literature revealed seven more cases: four in the head and neck, and one each in the skin, the lymph node, and the lung. IgM was the most frequently crystallizing immunoglobulin (four of seven) and all had kappa light chains. The lesion needs to be differentiated from neoplastic and nonneoplastic histiocytic and lymphoplasmacytic disorders. The difference with bronchial mucosa-associated lymphoid tissue lymphoma and marginal zone lymphoma is, perhaps, semantic. PMID- 9737250 TI - Epithelioid angiosarcoma arising in a surgically constructed arteriovenous fistula: a rare complication of chronic immunosuppression in the setting of renal transplantation. AB - Immunosuppression in the setting of solid organ transplantation is associated with the development of a variety of malignant tumors, most commonly squamous carcinomas and non-Hodgkin's lymphomas. Sarcomas, apart from Kaposi's sarcoma, are relatively infrequent. We recently encountered a 71-year-old man with chronic renal failure, treated by allograft kidney transplantation, who developed a high grade epithelioid angiosarcoma at the site of a nonfunctioning arteriovenous fistula, previously constructed for hemodialysis. At diagnosis, the patient had numerous satellite nodules of angiosarcoma involving the distal skin, soft tissues, and bones. After a below-elbow amputation, there was a rapid local recurrence at the amputation stump. Currently, the patient is alive with numerous pulmonary metastases, 6 months after amputation. A literature review identified three recently reported identical cases of epithelioid angiosarcoma arising in nonfunctioning arteriovenous fistulae. All three patients had been treated by kidney transplantation for renal failure, suggesting a possible causal association between these events. We performed polymerase chain reaction for human herpes virus 8, the recently recognized herpes virus proposed as a major etiologic agent of Kaposi's sarcoma, and possibly some conventional angiosarcomas, but we failed to identify any viral DNA within the tumor. PMID- 9737251 TI - Telepathology. PMID- 9737252 TI - Molecular alterations in salivary gland tumors. PMID- 9737253 TI - Fragments of artifactually created tissue intraoperatively retrieved from pericardial cavity. PMID- 9737254 TI - Radiation exposure, thyroid cancer and surgeons. PMID- 9737255 TI - 'Needlescopic' (mini-laparoscopic) surgery: necessary or unnecessary? PMID- 9737256 TI - Supracricoid laryngectomy: a significant advance in the management of laryngeal cancer. AB - Supracricoid laryngectomy is a new development in the treatment of laryngeal cancer in Australia. It allows the removal of both vocal cords, vestibular folds, and the entire thyroid cartilage including the paraglottic space without the loss of laryngeal function. This technique may offer a significant advance in the treatment of squamous cell carcinoma of the larynx. This paper reviews the surgical management of laryngeal cancer and discusses this new technique with modifications, indications for its use and the advantages of supracricoid laryngectomy. Case reports of seven patients operated on in our institution are included. PMID- 9737257 TI - Evaluation of thyroid shields for reduction of radiation exposure to orthopaedic surgeons. AB - BACKGROUND: A perceived increase in the incidence of thyroid carcinoma in orthopaedic surgeons prompted an assessment of the use and value of thyroid shields in the operating theatre. METHODS: The radiation exposure to the thyroid area of 19 orthopaedic trainees was measured over a 3-month period, while they were operating. The results were correlated with thyroid function tests and the number of emergency operative cases performed. RESULTS: Thirteen trainees received radiation exposure within the guidelines set for the general population. Two trainees received exposure above this but within the guidelines set for occupational exposure. A thyroid shield reduced radiation exposure of the neck in one trainee by a factor of 13. The availability and usage of thyroid shields was low: only seven out of 13 trainees used shields. CONCLUSIONS: A thyroid shield should be worn by orthopaedic surgeons if radiation is used during the operative procedure. PMID- 9737258 TI - Electrochemically induced hepatic necrosis: the next step forward in patients with unresectable liver tumours? AB - BACKGROUND: The treatment of patients with unresectable liver tumours remains an unsolved clinical problem. Several methods of locoregional treatment have been developed. These methods rely mainly on direct thermal or chemical insults and consequently have their own inherent limitations in clinical usage. The 'ideal' treatment would combine the direct cytotoxic effects of chemical treatments with the relative predictability of thermal insults, without the associated complications. This study aims to investigate whether the direct chemical effect of electrolytic hepatic necrosis is associated with any heating effect, and if so, whether the temperature change is dose-dependent. METHODS: An electrolytic 'dose' sufficient to induce a localized zone of hepatic necrosis was delivered to the livers of rats and pigs via implanted platinum electrodes. RESULTS: The results showed that there was no significant temperature increase at low current levels (2-4 mA) in the rat liver. In the pig, there was a significant (P < 0.01) increase in temperature of 4.2 degrees C during electrolysis, when delivered at between 20 and 50 mA. However, such a small increase in temperature would have been insufficient to cause appreciable thermal necrosis. CONCLUSIONS: This study demonstrates that electrolysis-induced hepatic necrosis is produced without an increase in temperature; clearly cell death results from the direct effects of cytotoxic electrode products and an alteration of intracellular pH. Consequently, it is likely that as a method for ablating liver tumours, electrolysis should be associated with fewer complications than other forms of locoregional treatment. PMID- 9737259 TI - Pelviureteric obstruction in children: conventional pyeloplasty is superior to endo-urology. AB - BACKGROUND: Hydronephrosis secondary to pelviureteric junction (PUJ) obstruction is common in infancy and childhood. Pyeloplasty has until recently been the accepted method of management, but alternative endo-urological techniques have evolved in the last decade. METHODS: Published results of conventional pyeloplasty for primary PUJ obstruction in children were compared with published results of endo-urological procedures. RESULTS: Sixty-six pyeloplasties were performed in 61 children in a 6-year period. During a similar period, 63 primary endo-urological procedures were reported in the literature. The success rate after pyeloplasty was 95.5% compared with 65% after endo-urology. CONCLUSIONS: Conventional pyeloplasty is superior to endo-urology and should remain the gold standard for the treatment of primary PUJ obstruction in children. PMID- 9737260 TI - Rotationplasty of the lower limb for childhood osteosarcoma of the femur. AB - BACKGROUND: Surgical management of osteosarcoma of the limb in childhood often involves extensive surgery. Intervention ranges from local resection to amputation depending on the level and the extent of the disease process. However, where the disease is extensive the level of amputation may preclude effective prosthetic use. In these circumstances resection of the diseased segment of the limb, i.e. an intercalary amputation and reconstruction with rotation of the tibial component of the limb can provide function equivalent to a below-knee amputee. METHODS: To examine the role of rotationplasty of the lower limb for childhood osteosarcoma of the femur, a retrospective study was carried out which examined the function of five patients who had been treated by a tibial rotationplasty. The records of two patients who had died from metastatic disease 2 and 4 years after surgery were examined. The three surviving patients were examined clinically and their clinical records and radiographs were reviewed. RESULTS: Of the two patients who died from metastatic disease, neither had local recurrence of the tumour, neurovascular complications, late derotations or psychological problems. In the three patients who survived, there had been no local recurrence of tumour or other complication and each has an excellent level of activity with below-knee function using a rotationplasty prosthesis. CONCLUSIONS: For children with osteosarcoma of the femur and with extensive disease that precludes complete limb preservation, resection and tibial rotationplasty provides function far superior to a high above-knee amputation or a hip disarticulation without the risk of local recurrence. PMID- 9737261 TI - Familial slipped capital femoral epiphysis: a report and considerations in management. AB - BACKGROUND: Familial inheritance of slipped capital femoral epiphysis (SCFE) is known. It has not been described in non-identical twins. A family where the mother and three of five siblings developed SCFE were investigated and managed. METHODS: Anthropometric measurement consisted of height-weight ratios. Serum sex hormone levels and bone Gla Protein was measured. Bone mineral densities were evaluated. RESULTS: The affected siblings had higher bodyweight percentiles. Other investigations were within normal limits. CONCLUSION: The unfavourable height-to-weight ratio was one of the mainstays in developing a management protocol for all siblings. The management protocol developed for the family is discussed. PMID- 9737262 TI - A study of the risk of strangulation and obstruction in groin hernias. AB - BACKGROUND: Complications that develop in groin hernias, such as irreducibility and obstruction, with or without strangulation may make an easily treatable condition a life-threatening one. Identification of risk factors that may predict development would help place the patient in a high-risk group. Priority admission and early elective surgery for such a patient would avoid significant mortality and morbidity. METHODS: This is a 10-year combined prospective and retrospective study of children and adults. Records of complicated groin hernias were identified from July 1985 to July 1995 from the outpatient department and available inpatient medical records. The same number of controls of simple uncomplicated hernias were then chosen using random number tables from among the large number belonging to the same time period. These two groups were then compared and analysed using statistical methods for age, sex, side of hernia, site of hernia (inguinal/femoral), duration of hernia, length of the waiting list for elective surgery, and contents of the hernial sac along with some other parameters to identify patients with high-risk factors. RESULTS: Age was found to be a significant risk factor and predicted complications in both elderly adults and very young children. Sex of the patient (male) and side of hernia (right) were significant risk factors in children only. Site of hernia was an important risk factor and adults with femoral hernia were most likely to experience complications. Duration of hernia for less than a year proved to be the most important risk factor for both children and adults. The majority of patients with complicated hernias had not presented earlier in the outpatient department, which implies that most hernias that become complicated do so within a very short time before patient referral. Mortality was high in patients with coexisting diseases, while morbidity was affected by viability of contents of the hernial sac which in turn was directly affected by duration of irreducibility or delay in presentation. CONCLUSIONS: The risk factors useful in predicting complications in an adult patient with groin hernia were age (older age group), duration of hernia (short duration), type of hernia (femoral more than inguinal) and coexisting medical illness. In children, the risk factors were age (very young), gender (male), short duration of hernia and side (right side). PMID- 9737263 TI - Primary reconstruction after extensive chest wall resection. AB - BACKGROUND: Chest wall resection and reconstruction has been proven to be a safe surgical procedure. This is particularly useful for breast cancer patients with chest wall recurrences or for those who first present with locally advanced cancer in the chest wall where there is both a large soft tissue and bony defect that need repair. In addition, many of these patients have had irradiation or chemotherapy, which can significantly impair wound healing. METHODS: Thirty-four patients underwent chest wall resection and primary reconstruction over an 8-year period. RESULTS: Twenty-three patients had breast carcinomas and six had breast and chest wall sarcomas. Of the breast carcinoma patients, 12 had local recurrences and 11 presented with locally advanced primary disease. Bony resection of the chest wall was required in 16 (47%) cases. Thirty myocutaneous flaps (18 rectus abdominis, four pectoralis major, eight latissimus dorsi) and three omental flaps were used for reconstruction. One required a deltovertebral skin flap. Skeletal reconstruction was necessary in four cases. All except one (97%) achieved primary wound healing. There was one mortality (3%) and three patients required further surgery for complications that were related to the reconstruction. Post-resection metastases occurred in 13 (42%) patients and only 2 (6%) had local recurrences. The 2-year survival rate was 78% with a mean survival time of 25.5 months. CONCLUSIONS: Primary reconstruction for curative or palliative purposes is a useful and safe surgical procedure for patients with recurrent or locally advanced chest malignancies after extensive chest wall resection. Pedicled myocutaneous flap is the preferred option for skeletal and soft-tissue coverage. PMID- 9737264 TI - Functional popliteal entrapment syndrome. AB - BACKGROUND: Classic popliteal artery entrapment is caused by the abnormal relationship between the popliteal artery and the medial head of the gastrocnemius, resulting in repetitive arterial compression and trauma. There is, however, a distinct subset of calf claudicants who have an anatomically normal popliteal fossa but can occlude the popliteal artery by repetitive vigorous exercise which involves active plantar flexion with or without extension at the knee joint. METHODS: Eight patients who led a vigorous athletic lifestyle were evaluated with duplex scan and biplane angiogram after being referred for bilateral calf claudication. They were found to have significant stenosis or occlusion of the popliteal artery with active plantar flexion. All patients had transection of the medial head of the gastrocnemius muscle with release of any vascular bands tethering the popliteal artery. RESULTS: Seven of the eight patients had complete relief. One patient noticed return of claudication at long distances, but a postoperative angiogram was normal. In all patients postoperative duplex scan showed no stenosis or occlusion of the popliteal vessels with the foot in active plantar flexion and the knee in extension. CONCLUSIONS: Functional popliteal artery entrapment is becoming a significant cause of disabling claudication in young athletic individuals and needs to be diagnosed accurately for appropriate treatment. This condition is becoming well known with the incorporation of sports in the daily routine of most young people. PMID- 9737265 TI - Improving the ease and safety of laparoscopy: a technique for open insertion of the umbilical trocar. AB - BACKGROUND: Sharp cannulation of the abdominal cavity for laparoscopy occasionally causes life-threatening injury to major vessels. METHODS: A technique of open cannulation is described to minimize this risk. RESULTS: In over 2000 general surgical laparoscopies performed the Flinders Medical Centre, there were no major vessel injuries and only one bowel injury (prior to laparotomy). CONCLUSIONS: We advocate the use of open technique to minimize injuries caused by abdominal cannulation for laparoscopy. PMID- 9737266 TI - The tensor fasciae latae myocutaneous flap closure of major chest and abdominal wall defects. AB - BACKGROUND: The usual methods of closure of major chest and abdominal wall defects have significant disadvantages. Skin grafts provide no structural support and result in incisional hernias. Synthetic mesh requires skin cover and is prone to infection and wound breakdown. The tensor fasciae latae (TFL) myocutaneous flap offers skin cover and a semi-rigid fascial layer. We document our unit's experience in pedicled and free TFL flaps. METHODS: The TFL flap closure of trunk defects was undertaken in 10 patients between August 1989 and April 1997. All cases were not amenable to primary closure and repair with synthetic mesh or skin grafts. RESULTS: The defect was satisfactorily repaired in all cases without subsequent herniation. The closure techniques using a pedicled TFL flap and a TFL flap for a free-tissue transfer are described. CONCLUSIONS: We conclude that the TFL flap is the method of choice for repairs of major truncal defects. PMID- 9737267 TI - A preliminary exploration of the interactional skills of trainee surgeons. AB - BACKGROUND: The interactional skills of 21 surgical trainees were studied in the areas of breaking bad news to patients and preparing patients for potentially threatening medical procedures. When compared to the established guidelines for dealing with these issues, the trainees performed poorly. METHODS: Trainees were videotaped using clinical histories delivered by simulated patients in simulated consulting rooms. All videos were scored on standard rating scales where the criteria for rating the specific interactional skills were adopted from existing guidelines. RESULTS: The proportion of trainees who could meet the guidelines when breaking bad news was low. The proportion of trainees who met the guidelines were: closing the the consultation (0%), provided patient information about prognosis (43%) and treatment (38%) and when they gave support (10%). There were also low numbers of trainees who could meet the guidelines when they prepared patients for potentially threatening procedures. Ten per cent of trainees followed the guidelines when providing information, 25% could establish treatment goals, 35% could give a prognosis and 0% could deal appropriately with psychosocial issues or close the consultation appropriately. CONCLUSIONS: It is concluded that there is a need for further formal training in interactional skills as part of surgical training. PMID- 9737268 TI - Amputation through the ages: the oldest major surgical operation. AB - Amputation has been practised since neolithic times for punitive, therapeutic and ritualistic reasons. Until the development and adoption of general anaesthesia, it was the most major operation to which a surgeon could aspire and a speedy technique was essential. Many famous names have been associated with this operation in both military and civilian surgery. PMID- 9737270 TI - Bilateral iliofemoral venous thrombosis: a rare presentation of ruptured aortic aneurysm. PMID- 9737269 TI - Gynaecomastia presenting as fibroadenomatoid tumours of the breast in a renal transplant recipient associated with cyclosporin treatment. PMID- 9737271 TI - Management of sigmoid colon intussusception presenting through the anus. PMID- 9737272 TI - Viagra's licence and the internet. PMID- 9737273 TI - Congenital adrenal hyperplasia--a continuum of disorders. PMID- 9737274 TI - Appropriate technology for cataract surgery. PMID- 9737275 TI - Selenium and risk of prostate cancer. PMID- 9737276 TI - Documentation of clinical benefit of specific aminoacid nutrients. PMID- 9737277 TI - Measuring the burden of disease. PMID- 9737278 TI - Dropped-head and bent-spine syndromes; axial myopathies? PMID- 9737279 TI - Changes in physical fitness and changes in mortality. AB - BACKGROUND: Point estimates of physical fitness give important information on the risk of death in healthy people, but there is little information available on effects of sequential changes in physical fitness on mortality. We studied this latter aspect in healthy middle-aged men over a total follow-up period of 22 years. METHODS: 2014 healthy men aged 40-60 years had a bicycle exercise test and clinical examination, and completed a questionnaire in 1972-75 (survey 1). This was repeated for 1756 (91%) of 1932 men still alive by Dec 31, 1982 (survey 2). The exercise scores were adjusted for age. The change in exercise scores between surveys was divided into quartiles (Q1=least fit, Q4=fittest). An adjusted Cox's proportional hazards model was used to study the association between changes in physical fitness and mortality, with the Q1 men used as controls. FINDINGS: By Dec 31, 1994, 238 (17%) of the 1428 men had died, 120 from cardiovascular causes. There were 37 deaths in the Q4 group (19 cardiovascular); their relative risks of death were 0.45 (95% CI 0.29-0.69) for any cause and 0.47 (0.26-0.86) for cardiovascular causes. There was a graded, inverse relation between changes in physical fitness and mortality irrespective of physical fitness status at survey 1. INTERPRETATION: Change in physical fitness in healthy middle-aged men is a strong predictor of mortality. Even small improvements in physical fitness are associated with a significantly lowered risk of death. If confirmed, these findings should be used to influence public health policy. PMID- 9737280 TI - Prevalence of thyroid autoantibodies in children and adolescents from Belarus exposed to the Chernobyl radioactive fallout. AB - BACKGROUND: The long-term effects of ionising radiation, including radioiodine, on thyroid function are not well known. We compared thyroid immunity and function in two groups of children from Belarus, one of whom was exposed to the radioactive fallout of Chernobyl. METHODS: We measured serum free thyroxine 4 (free T4), free T3, and thyrotropin hormone (TSH) and the prevalence of thyroid autoantibodies (antithyroglobulin and antithyroperoxidase), in 287 children or adolescents living in Hoiniki (average caesium contamination of 5.4 Ci/km2). We also studied 208 children and adolescents living in Braslav (average contamination <0.1 Ci/km2), who were age 12 years or less at the time of the Chernobyl accident. FINDINGS: The prevalence of antithyroglobulin or antithyroperoxidase, or both, was significantly higher (p=0.0001) in individuals living in Hoiniki (56 [19.5%] of 287) than in those living in Braslav (eight [3.8%] of 208). In both villages, no sex differences were found in the antibody prevalence before age 13 years. Thereafter, a significantly higher prevalence of thyroid autoantibodies was found in girls from Hoiniki. The increase in the prevalence of circulating antibodies in the contaminated group was already apparent in individuals who, at the time of the accident, were in utero or newborn (15.7%), and was even more pronounced in children of 9 years or more (35.1%). No major alterations of serum FT-4, FT-3, or TSH were found. INTERPRETATION: 6-8 years after the Chernobyl accident, a significant increase in thyroid autoimmunity was found in children exposed to radioactive fallout. Pubertal age in girls is a risk factor for increased prevalence of thyroid autoimmunity. The autoimmune phenomena are limited to an increased prevalence of circulating thyroid autoantibodies without evidence of significant thyroid dysfunction. The future development of clinically relevant thyroid autoimmune diseases, especially hypothyroidism, is a possibility. PMID- 9737281 TI - West Nile encephalitis epidemic in southeastern Romania. AB - BACKGROUND: West Nile fever (WNF) is a mosquito-borne flavivirus infection endemic in Africa and Asia. In 1996, the first major WNF epidemic in Europe occurred in Romania, with a high rate of neurological infections. We investigated the epidemic to characterise transmission patterns in this novel setting and to determine its origin. METHODS: Hospital-based surveillance identified patients admitted with acute aseptic meningitis and encephalitis in 40 Romanian districts, including Bucharest. Infection was confirmed with IgM capture and indirect IgG ELISAs. In October, 1996, we surveyed outpatients in Bucharest and seven other districts to estimate seroprevalence and to detect infected patients not admitted to hospital. We also measured the rates of infection and seropositivity in mosquitoes and birds, respectively. RESULTS: Between July 15 and Oct 12, we identified 393 patients with serologically confirmed or probable WNF infection, of whom 352 had acute central-nervous-system infections. 17 patients older than 50 years died. Fatality/case ratio and disease incidence increased with age. The outbreak was confined to 14 districts in the lower Danube valley and Bucharest (attack rate 12.4/100000 people) with a seroprevalence of 4.1%. The number of mild cases could not be estimated. WN virus was recovered from Culex pipiens mosquitoes, the most likely vector, and antibodies to WN virus were found in 41% of domestic fowl. INTERPRETATION: The epidemic in Bucharest reflected increased regional WNF transmission in 1996. Epidemics of Cx pipiens-borne WNF could occur in other European cities with conditions conducive to transmission. PMID- 9737282 TI - Randomised trial of glutamine-enriched enteral nutrition on infectious morbidity in patients with multiple trauma. AB - BACKGROUND: Infections are an important cause of morbidity and mortality in patients with multiple trauma. Studies in both animals and human beings have suggested that glutamine-enriched nutrition decreases the number of infections. METHODS: Patients with multiple trauma with an expected survival of more than 48 h, and who had an Injury Severity Score of 20 or more, were randomly allocated glutamine supplemented enteral nutrition or a balanced, isonitrogenous, isocaloric enteral-feeding regimen along with usual care. Each patient was assessed every 8 h for infection, the primary endpoint. Data were analysed both per protocol, which included enteral feeding for at least 5 days, and by intention to treat. FINDINGS: 72 patients were enrolled and 60 received enteral feeding (29 glutamine-supplemented) for at least 5 days. Five (17%) of 29 patients in the glutamine-supplemented group had pneumonia compared with 14 (45%) of 31 patients in the control group (p<0.02). Bacteraemia occurred in two (7%) patients in glutamine group and 13 (42%) in the control group (p<0.005). One patient in the glutamine group had sepsis compared with eight (26%) patients in the control group (p<0.02). INTERPRETATION: There was a low frequency of pneumonia, sepsis, and bacteraemia in patients with multiple trauma who received glutamine-supplemented enteral nutrition. Larger studies are needed to investigate whether glutamine-supplemented enteral nutrition reduces mortality. PMID- 9737283 TI - Randomised trial of management of hypertensive pregnancies by Korotkoff phase IV or phase V. AB - BACKGROUND: There is debate about whether diastolic blood pressure should be recorded as the fourth (muffling, K4) or fifth (disappearance, K5) Korotkoff sound in pregnancy. We compared maternal and fetal outcomes and the likelihood that episodes of severe hypertension would be recorded when hypertensive pregnancies were managed according to either K4 or K5. METHODS: 220 pregnant women with diastolic hypertension (K4 > or =90 mm Hg) after the 20th week of gestation were enrolled in a prospective randomised study at two obstetric units in Australia; they were randomly assigned management with K4 (n=103) or K5 (n=117) for the remainder of the pregnancy. Clinical management was according to a uniform department protocol. Analysis was by intention to treat. All the women completed the trial. FINDINGS: An episode of severe hypertension (systolic > or =170 mm Hg, diastolic > or =110 mm Hg, or both) was more likely to be recorded with use of K4 than with use of K5 (39 [38%] vs 30 [26%] women, p=0.051), mainly because of a greater likelihood that severe diastolic hypertension would be recorded (34 [33%] vs 20 [17%], p=0.006). The frequency of severe systolic hypertension and simultaneous severe systolic and diastolic hypertension did not differ between groups. Pregnancy was prolonged by an average of 2 weeks in both groups, and there were no significant differences between the groups in laboratory data, requirements for antihypertensive treatment, birthweight, fetal growth retardation, or perinatal mortality. There was no eclampsia or significant maternal morbidity in either group. INTERPRETATION: A change from use of K4 to K5 would mean that one fewer case of severe diastolic hypertension would be recorded for every six hypertensive pregnancies, but all other episodes of severe hypertension would be recorded with similar frequency. Since the K4/K5 difference is smaller in hypertensive than in normotensive pregnant women and since K5 is closer to the actual intra-arterial pressure and more reliably detected, universal adoption of K5 to record diastolic blood pressure in hypertensive pregnancy should be considered. PMID- 9737284 TI - A new treatment for toxic megacolon. PMID- 9737285 TI - HIV-1-associated nephropathy and response to highly-active antiretroviral therapy. PMID- 9737286 TI - Neuroleptics in progressive structural brain abnormalities in psychiatric illness. PMID- 9737287 TI - Alcohol, drinking, illicit drug use, and stress in junior house officers in north east England. PMID- 9737288 TI - Ultrasonography of contralateral veins in patients with unilateral deep-vein thrombosis. PMID- 9737289 TI - Women's magazines and tobacco in Europe. PMID- 9737291 TI - Detection of Helicobacter pylori DNA in houseflies (Musca domestica) on three continents. PMID- 9737290 TI - Tar concentrations in cigarettes and carcinogen content. PMID- 9737292 TI - Disappearance of an Epstein-Barr virus-positive post-transplant plasmacytoma with reduction of immunosuppression. PMID- 9737293 TI - Mamillary body abnormalities in schizophrenia. PMID- 9737294 TI - Etruscan wombs. PMID- 9737295 TI - Scientists identify a new HIV-1 group. PMID- 9737296 TI - Beta-blockers given the go-ahead for treatment of heart failure. PMID- 9737297 TI - Combing the oceans for new therapeutic agents. PMID- 9737298 TI - UK government responds to variant-CJD news and waiting lists. PMID- 9737299 TI - Health Minister Zuma under tobacco fire in South Africa. PMID- 9737300 TI - Ireland's success with HIV pregnancies. PMID- 9737301 TI - Caring for Japan's elderly--mission impossible? PMID- 9737302 TI - Carcinoid tumour. AB - Carcinoid tumours are often indolent asymptomatic tumours. However, a small but significant proportion are malignant and difficult to manage. Multiple endocrine neoplasia type 1 (MEN-1) may be associated with carcinoid tumours and should therefore be considered in the investigation of these patients. This review puts into context the use of newer imaging modalities, including octreotide scintigraphy. The therapeutic treatment options are discussed, including the use of octreotide, the role of receptor-targeted therapy, hepatic-artery embolisation, and the arguments against chemotherapy. We review the need for careful patient selection when considering curative and palliative surgery, including liver transplantation. We conclude that there are now better diagnostic tools and therapeutic options available for those patients with malignant carcinoid tumours, and that these patients are best managed by a multidisciplinary approach. Earlier detection and treatment of these tumours should lead to improved quality of life and survival, which, ideally, should be assessed in formal trials. PMID- 9737303 TI - In search of a medical artist. PMID- 9737304 TI - Should the mission of epidemiology include the eradication of poverty? PMID- 9737305 TI - One fortunate old man. PMID- 9737306 TI - Shaken babies. PMID- 9737307 TI - Shaken babies. PMID- 9737308 TI - Shaken babies. PMID- 9737309 TI - Pituitary insufficiency. PMID- 9737310 TI - Pituitary insufficiency. PMID- 9737311 TI - Pituitary insufficiency. PMID- 9737312 TI - Antithrombotics for left-ventricular impairment. PMID- 9737313 TI - Non-steroidal anti-inflammatory drugs and metformin. PMID- 9737314 TI - Smoking and risk of Alzheimer's disease. PMID- 9737315 TI - Smoking and risk of Alzheimer's disease. MIRAGE Study Group. PMID- 9737316 TI - Tamoxifen in hepatocellular carcinoma. PMID- 9737317 TI - Aortic stenosis and ACE inhibitors. PMID- 9737318 TI - Aortic stenosis and ACE inhibitors. PMID- 9737319 TI - Metabolic abnormalities and use of HIV-1 protease inhibitors. PMID- 9737320 TI - Transmission of tumours by liver transplantation. PMID- 9737321 TI - Cor pulmonale and tumour cell microemboli. PMID- 9737322 TI - Silicone breast implants. PMID- 9737323 TI - Long-acting oily chloramphenicol for meningococcal meningitis. PMID- 9737324 TI - Mibefradil: the sole exception. PMID- 9737325 TI - Jehovah's Witnesses and blood transfusions. PMID- 9737326 TI - "I will come for the benefit of the sick". PMID- 9737327 TI - 2C-B: a new psychoactive phenylethylamine recently discovered in Ecstasy tablets sold on the Swiss black market. AB - This study sought to identify, by means of several analytical methods (GC-MS, HPLC-DAD, CE-DAD, FTIR, and NMR), 4-bromo-2,5-dimethoxyphenethylamine (2C-B), which was found in two sets of tablets obtained from the Swiss black market. Unequivocal identification of 2C-B was only achieved by a combination of mass spectrometric and NMR analysis. Quantitation of 2C-B was performed by HPLC-DAD and CE-DAD. The amounts of 2C-B found in the tablets (3-8 mg) were in the range of the minimum quantity required to induce the effects characteristic of this drug. PMID- 9737328 TI - Cocaine and cocaethylene binding to human liver. AB - The binding of cocaine (COC) and cocaethylene (CE) to whole human liver homogenates in vitro was studied by equilibrium dialysis. Drugs were measured by high-pressure liquid chromatography. Up to 32% of COC and up to 43% of CE were bound. Scatchard analysis suggested a high-affinity, low-capacity binder for both COC (Ka, 4.69 x 10(4) L/mol; Bo, 1.08 x 10(-5) mol/L) and CE (Ka, 4.38 x 10(4) L/mol; Bo, 1.54 x 10(-5) mol/L). In addition, low-affinity, high-capacity binders for COC (Ka, 2.93 x 10(3) L/mol; Bo, 1.32 x 10(-4) mol/L) and CE (Ka, 6.50 x 10(3) L/mol; Bo, 1.11 x 10(-4) mol/L) were noted. Finally, for both compounds, very low-affinity, high-capacity binding, which was likely nonspecific in nature, was defined as follows: COC, Ka, 8.00 x 10(2) L/mol; Bo, 5.45 x 10(-4) mol/L and CE, Ka, 2.10 x 10(3) L/mol; Bo, 3.71 x 10(-4) mol/L. The binding profiles of COC and CE in liver were compared with those in human serum and placenta studied previously by this laboratory. PMID- 9737329 TI - Simultaneous extraction of selected benzodiazepines and benzodiazepine glucuronides from urine by immunoadsorption. AB - A rapid and selective cleanup procedure based on immunoadsorption is described for the simultaneous extraction of diazepam and its free or glucuronidated metabolites nordiazepam, temazepam, and oxazepam from urine. The method can also be used for the extraction of lorazepam and lorazepam-glucuronide. Because the samples do not have to be hydrolyzed before extraction, valuable information is preserved. With the exception of lorazepam-glucuronide, recoveries between 86 and 100% were obtained at spiking levels up to 200 ng of benzodiazepine or glucuronide per milliliter of urine. Using methanol/water (90:10, v/v) as an eluent, the immunoadsorber could be used at least 20 times. High-performance liquid chromatograms of urine samples from patients receiving low therapeutic dosages of diazepam or lorazepam are shown to demonstrate the high purity of the extracts. PMID- 9737330 TI - A new specific method to detect cyanide in body fluids, especially whole blood, by fluorimetry. AB - This study shows a simple, rapid, and specific method for the quantitative determination of cyanide ion in body fluids, especially blood, by fluorimetry. It is based upon the transformation of cyanide ion into hydrocyanic acid, which then reacts with 2,3-naphthalenedialdehyde and taurine in a self-contained system. The 1-cyano-2-benzoisoindole derivate thus formed is suitable for fluorimetric measurement (lambdaEX = 418 nm; lambdaEM = 460 nm). The fluorescence intensity can be determined by spectrophotometry or by high-performance liquid chromatography (HPLC) with fluorescence detection. The detection limit is 0.002 microg/mL. Linearity was excellent from 0.002 to 1 microg/mL for spectrophotometry and from 0.002 to 5 microg/mL for HPLC with fluorescence detection. The coefficient of variation for repeatability was 8% or less. Thiocyanate and sulfide did not interfere, even at high concentrations (200 microg/mL). The method was applicable to whole blood, so it should be suitable for both clinical and forensic purposes. PMID- 9737331 TI - Development and validation of a screening method for DES, zeranol, and beta zearalanol in bovine urine by HRGC-MS and evaluation of robustness for routine survey of the Brazilian herd. AB - A method and evaluated for screening and confirmation of diethylstilbestrol (DES), alpha- and beta-zearalanol in bovine urine was developed. The residues were extracted from urine by C18 cartridges and purified on alumina columns. For screening and confirmation purposes, the anabolic derivatives were analyzed by gas chromatography-mass spectrometry after derivatization with BSTFA + 1% TMCS or a solution of PFPA/acetone (1:2, v/v), respectively. The recovery of most analytes for the whole procedure was higher than 96%, with a detection limit of 0.5 ppb. This procedure is being routinely applied to the Brazilian National Program for the Control of Residues in Meat (PNCRBC). PMID- 9737332 TI - Isotope dilution gas chromatographic-mass spectrometric measurement of tricyclic antidepressant drugs. Utility of the 4-carbethoxyhexafluorobutyryl derivatives of secondary amines. AB - Stable isotope dilution gas chromatographic-mass spectrometric (GC-MS) measurement of tricyclic antidepressants (TCA) is a useful alternative to high performance liquid chromatography (HPLC) methods when interfering substances prevent accurate quantitation by HPLC. For satisfactory GC-MS analysis, secondary amine TCA must be derivatized. Commonly employed trifluoroacetyl and heptafluorobutyryl derivatives are relatively unstable and cause rapid deterioration of capillary GC columns. Therefore we examined 4 carbethoxyhexafluorobutyryl chloride (CHFB-CI) as an alternative derivatizing agent and developed a stable isotope dilution GC-MS method employing ring-labeled [2H4]-desipramine and [2H4]-imipramine internal standards, which permits measurement of desipramine, nortriptyline, imipramine, and amitriptyline in plasma samples containing one or all of these analytes. The GC-MS assay is linear for each analyte from the lower limit of quantitation (25 ng/mL) up to 1500 ng/mL and correlates well with HPLC measurements. The GC-MS analytic coefficient of variation was 9.7 +/- 1.3% for all analytes considered together. Although interferences are observed in the HPLC assay, thioridazine, perphenazine, cyclobenzaprine, and norcyclobenzaprine do not interfere with GC-MS measurements of the TCA examined here. The stability of the CHFB derivative of secondary amine TCA was found to be superior to that of the trifluoroacetyl derivatives of these compounds. PMID- 9737333 TI - Identification of metabolites of nerve agent VX in serum collected from a victim. AB - A human serum sample collected from a victim of the Osaka VX incident was analyzed according to our developed technique for metabolites of VX. Gas chromatography-mass spectrometry (GC-MS) in full-scan electron impact and chemical ionization modes were used, and, for more reliable confirmation, GC-MS MS was also employed. In the serum sample, both ethyl methylphosphonic acid and 2 (diisopropylamino-ethyl)methyl sulfide were detected. These results indicated that the techniques using GC-MS and GC-MS-MS were applicable to biological samples such as serum. These results also provide the first documented, unequivocal identification of the specific metabolites of VX in victim's serum and, furthermore, clarify a part of the metabolic pathway of VX in the human body. PMID- 9737334 TI - Improved solid-phase extraction of methadone and its two major metabolites from whole blood. AB - A solid-phase extraction (SPE) method for the efficient extraction of methadone and its two major metabolites, 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine and 2-ethyl-5-methyl-3,3-diphenyl-1-pyrroline, from whole blood is described. The procedure combines extraction on Isolute Confirm HCX mixed-mode SPE columns and gas chromatographic-mass spectrometric analysis with deuterated methadone as the internal standard. The optimum extraction conditions for all three analytes were determined using spiked whole blood. The developed method is easier and faster than current liquid-liquid extraction (LLE) procedures and produces cleaner extracts. Calibration curves were linear from 0 to 600 ng/mL (r2 > 0.99) with recoveries greater than 90% for all three analytes. The concentrations of methadone and its metabolites in postmortem blood were determined in fatal cases using the developed SPE method and were found to compare well with results obtained using LLE. PMID- 9737335 TI - False-positive LSD testing in urine samples from intensive care patients. AB - Unexpected positive results for lysergic acid diethylamide (LSD) were found in urine samples from 12 patients in an intensive care unit in a routine screening using the CEDIA DAU assay. None of these test results could be confirmed by high performance liquid chromatography analysis, but all samples contained the mucolytic drug ambroxol. Further studies demonstrated that ambroxol exhibits a significant cross-reactivity in the CEDIA DAU LSD assay. Therefore, positive LSD results obtained with the CEDIA DAU assay have to be critically evaluated, particularly during the cold season, when infections of the respiratory tract often result in more frequent use of mucolytic medications. PMID- 9737336 TI - Simple analysis of tetracyclic antidepressants in blood using headspace-solid phase microextraction and GC-MS. AB - A simple and sensitive method for analysis of three tetracyclic antidepressants, maprotiline, mianserin, and setiptiline, in human whole blood was developed using headspace-solid-phase microextraction (SPME) and gas chromatography-mass spectrometry (GC-MS). A vial containing a blood sample, sodium hydroxide, and imipramine as an internal standard was heated at 120 degrees C. The extraction fiber of the SPME was exposed for 45 min in the headspace of the vial. The compounds absorbed on the fiber were desorbed by exposing the fiber in the injection port of a GC-MS. The calibration curves, using an internal standard method, demonstrated good linearity throughout the concentration range from 0.005 to 5.0 microg/g for mianserin and setiptiline and from 0.025 to 25 microg/g for maprotiline. No interferences were found, and the time for analysis was 60 min for one sample. In addition, this proposed method was applied to a medicolegal case in which the cause of death was suspected to be acute setiptiline poisoning. Setiptiline was detected in the left and right heart blood samples of the victim at concentrations of 1.77 and 0.78 microg/g, respectively. PMID- 9737337 TI - Nitrite contamination in urine collection kit component-absorbent sheet. PMID- 9737338 TI - Evaluation of DRI drug-of-abuse reagents on the Hitachi 911. PMID- 9737340 TI - Fourier transform infrared microspectroscopic analysis of bones of osteocalcin deficient mice provides insight into the function of osteocalcin. AB - Osteocalcin, the gamma-carboxyglutamic acid-containing protein, which in most species is the predominant noncollagenous protein of bone and dentin, has been postulated to play roles in bone formation and remodeling. Recently, genetic studies showed that osteocalcin acts as an inhibitor of osteoblast function. Based on von Kossa staining and measurement of mineral apposition rates in tetracycline-labeled bones, osteocalcin knockout animals were reported to have no detectable alterations in bone mineralization. To test the hypothesis that, in addition to regulating osteoblastic activity, osteocalcin is involved in regulating mineral properties, a more sensitive assay of mineralization, Fourier transform infrared microspectroscopy (FT-IRM) was used to study thin sections of femora of 4-week-, 6-month- (intact and ovariectomized), and 9-month-old wild type and osteocalcin-knockout mice. FT-IRM spectra provided spatially resolved measures of relative mineral and carbonate contents, and parameters indicative of apatite crystal size and perfection. No differences were detected in the mineral properties of the 4-week-old knockout and wild-type mice indicating that the mineralization process was not altered at this time point. Six-month-old wild type animals had higher mineral contents (mineral:matrix ratios) in cortical as compared with trabecular bones; mineral contents in knockout and wild-type bones were not different. At each age studied, carbonate:phosphate ratios tended to be greater in the wild-type as compared with knockout animals. Detailed analysis of the phosphate nu1,nu3 vibrations in the spectra from 6-month-old wild-type animals indicated that the crystals were larger/more perfect in the cortical as opposed to the trabecular bones. In contrast, in the knockout animals' bones at 6 months, there were no differences between trabecular and cortical bone in terms of carbonate content or crystallite size and perfection. Spectral parameters of the cortical and trabecular bone of the knockout animals resembled those in the wild-type trabecular bone and differed from wild-type cortical bone. In ovariectomized 6-month-old animals, the mineral content (mineral:matrix ratio) in the wild-type cortices increased from periosteum to endosteum, whereas, in the knockout animals' bones, the mineral:matrix ratio was constant. Ovariectomized knockout cortices had lower carbonate:phosphate ratios than wild-type, and crystallite size and perfection resembled that in wild-type trabeculae, and did not increase from periosteum to endosteum. These spatially resolved data provide evidence that osteocalcin is required to stimulate bone mineral maturation. PMID- 9737339 TI - Insulin receptor expression in primary and cultured osteoclast-like cells. AB - Skeletal growth is the net product of coordinated bone formation and resorption. Insulin is known to stimulate bone formation by actions on osteoblasts. It is not known whether insulin receptors are present on osteoclasts, or whether insulin regulates osteoclastic function. We present here immunocytochemical evidence of insulin receptor expression by mature mono- and multinucleated murine osteoclast like cells generated in vitro, and in primary neonatal rat and mouse osteoclasts. Radiolabeled studies indicated that progressive enrichment of osteoclast-like cells in coculture was associated with increased insulin binding. When osteoclast like cells generated in vitro were plated onto dentine slices, insulin dose dependently inhibited pit formation by up to 80%, suggesting a role for insulin in osteoclast function. These data are consistent with an effect of insulin on bone resorption in addition to those previously recognized on bone formation, actions that together result in net bone growth. PMID- 9737341 TI - Catecholamines stimulate the proliferation and alkaline phosphatase activity of MC3T3-E1 osteoblast-like cells. AB - A number of factors have been shown to influence osteoblastic proliferation, including fluoride. Recent observations suggest that heterotrimeric G proteins are probably involved in the mitogenic response induced by this agent, further suggesting a role of guanosine 5'-triphosphate (GTP)-binding protein-coupled receptors (GPCR) in the regulation of osteoblastic cell growth. We therefore explored what mitogenic factors known to activate GPCR can influence the replication of mouse osteoblast-like MC3T3-E1 cells. Among several candidates, epinephrine was found to be a potent mitogen for these cells, and its effect on the growth and differentiation of these cells was further investigated. Deoxyribonucleic acid (DNA) synthesis was dose dependently enhanced by this catecholamine in the concentration range of 1 nmol/L-10 micromol/L. Stimulation of DNA synthesis by catecholamines was in the order of epinephrine > norepinephrine >> isoproterenol, indicating that alpha adrenergic receptors mediated this cellular response. Further analysis with specific adrenergic receptor agonists and antagonists suggested that the mitogenic response induced by epinephrine in MC3T3-E1 cells is mediated by alpha1 adrenergic receptors. In addition to its effect on cell replication, epinephrine also enhanced alkaline phosphatase (ALP) activity in these cells but had little effect on collagen synthesis and osteocalcin production. As for the mitogenic response, the change in ALP activity was found to be mediated by alpha1 adrenergic receptors. Both effects of epinephrine on cell replication and ALP activity were markedly reduced by pretreatment of the cells with pertussis toxin (PTX), suggesting a role of Gi proteins. These effects were also completely blocked by pretreatment of the cells with 50 micromol/L genistein, a nonselective inhibitor of tyrosine kinase. In conclusion, the results indicate that epinephrine enhances replication and ALP activity of MC3T3-E1 osteoblast-like cells via alpha1 adrenergic receptors coupled to Gi proteins. The signaling mechanism probably involves a tyrosine phosphorylation mechanism. These observations suggest that PTX-sensitive G proteins are potent mediators of cell proliferation and ALP activity of osteoblast-like cells in response to factors acting through G protein-coupled receptors. PMID- 9737342 TI - Lymphoblastoid interferon-alpha downregulates parathyroid hormone (PTH)/PTH related peptide (PTHrP) receptor expression in human osteoblastic cells (Saos-2). AB - Interferon-alpha (IFN-alpha) is a pleiotropic cytokine that modulates the cellular functions of both osteoblastic and osteoclastic lineages. It remains unclear whether IFN-alpha regulates the expression of parathyroid hormone (PTH)/PTH-related peptide (PTHrP) receptor, which is a major target molecule regulating skeletal metabolism. In this study, we examined the effect of IFN alpha on the expression of PTH/PTHrP receptor in human osteoblastic cells (Saos 2). IFN-alpha inhibited the expression of PTH/PTHrP receptor gene in both a time- and dose-dependent manner. The mRNA level was decreased to 61.1% of that of the untreated control by 48 h treatment with 6000 U/mL of IFN-alpha. IFN-alpha also decreased cAMP response to PTH(1-34) in a dose-dependent manner and significantly depressed expression of the receptor protein. However, IFN-alpha did not exert any effect on other osteoblastic markers, such as alkaline phosphatase (ALP) activity, cAMP response to prostaglandin E2 (PGE2), and secretion of bone gla protein (BGP) and bone sialoprotein (BSP). Finally, IFN-alpha decreased PTH/PTHrP receptor mRNA to 60.7% that of control in the presence of actinomycin D. These data suggest that IFN-alpha downregulates the expression of PTH/PTHrP receptor and its signaling without affecting other osteoblastic markers, and that IFN alpha regulates its gene expression mainly by decreasing the stability of its mRNA. PMID- 9737343 TI - Phorbol ester-induced production of prostaglandin E2 from phosphatidylcholine through the activation of phospholipase D in UMR-106 cells. AB - To determine the effects of 12-O-tetradecanoylphorbol-13-acetate (TPA) on phospholipase D (PLD) activity in osteoblast-like UMR-106 cells, we used cells prelabeled with [3H] myristic acid or [3H] arachidonic acid, which were preferentially incorporated to phosphatidylcholine. The treatment of [3H] myristate-labeled cells with TPA in the presence of 1% ethanol caused a dose dependent formation of [3H] phosphatidylethanol (PEt), a product specific to PLD, suggesting an activation of this enzyme. Pretreatment of the cells with protein kinase C (PKC) inhibitors (GF109203X, staurosporine or H-7) abolished the TPA dependent formation of PEt. The PEt formation in response to TPA treatment was not observed after the pretreatment of the cells with TPA to downregulate PKC. These results suggest the involvement of PKC in the TPA-induced activation of PLD. With [3H] arachidonate-labeled cells, TPA treatment in the absence of ethanol resulted in the liberation of [3H] arachidonic acid, which was gradually converted to prostaglandin E2 (PGE2), but the accumulations of [3H] phosphatidic acid (PA) and [3H] diacylglycerol (DAG) were very small and temporary. In contrast, PA was linearly accumulated following TPA treatment, when the cells were pretreated with an inhibitor of phosphatidate phosphohydrolase (PAP), propranolol, with no accumulation of either DAG or arachidonic acid. The TPA treatment of the cells pretreated with a DAG lipase inhibitor, RHC-80267, caused the generation of DAG after a lag period of approximately 5 min, with a very small and temporary accumulation of PA. The TPA treatment of cells pretreated with a cyclooxygenase (COX) inhibitor, indomethacin, blocked the PGE2 production. The TPA-induced PGE2 production was not affected by the pretreatment of cells with a phospholipase A2 inhibitor, p-bromophenacylbromide, or with a phospholipase C inhibitor, D-609. TPA also stimulated PGE2 production in osteoblastic cells that were enzymatically isolated from adult rat calvaria, and the experiments with lipid metabolizing enzyme inhibitors gave the same profile of inhibition of TPA-induced PGE2 production as was observed in UMR-106 cells. These results suggest that PA formed as a consequence of the activation of PLD by TPA is rapidly converted to arachidonic acid via a PAP/DAG lipase pathway, followed by a gradual conversion of arachidonic acid to PGE2 by COX in both UMR 106 cells and isolated adult osteoblastic cells, and that neither phospholipase A2 nor phospholipase C is involved in the TPA-induced PGE2 production. To the best of our knowledge, this is the first report that shows that the activation of PKC in osteoblastic cells leads to the production of PGE2 via a PLD/PAP/DAG lipase/COX pathway. PMID- 9737344 TI - Effects of recombinant human bone morphogenetic protein-2 on differentiation of cells isolated from human bone, muscle, and skin. AB - We investigated the effects of recombinant human BMP-2 (rhBMP-2) on differentiation of cells isolated from human bone, muscle, and skin. Cells isolated from bones of six patients (HBM-1 to HBM-6), muscle from five patients (HM-1 to HM-5), and skin from three patients (HF-1 to HF-3) were used. rhBMP-2 had no effects on proliferation of two HBM cells, but had a stimulatory effect on three HM cell samples. rhBMP-2 stimulated both alkaline phosphatase (ALP) activity in all HBM cells and parathyroid hormone (PTH)-dependent cAMP production in three of the four HBM cell samples, although the magnitudes of these stimulatory effects differed among the cells tested. Although none of the HBM cells examined produced detectable amounts of osteocalcin in the absence of 1,25 (OH)2vitamin D3, they synthesized measurable amounts of osteocalcin in its presence. rhBMP-2 inhibited 1,25-(OH)2vitamin D3-dependent osteocalcin production in all of the HBM cell samples. Transplantation of HBM-6 cells with rhBMP-2 using diffusion chambers into the peritoneal cavity of athymic mice induced formation of cartilage and bone in the diffusion chambers, but neither cartilage nor bone was formed in chambers transplanted without rhBMP-2. rhBMP-2 also stimulated ALP activity in all of the HM and HF cell samples examined and PTH-dependent cAMP production in three of four HM cell samples. rhBMP-2 induced no osteocalcin production in any of the HM or HF cells in the presence of 1,25-(OH)2vitamin D3. rhBMP-2 markedly inhibited myotube formation by all of the HM cell samples. Transplantation of HM-4 cells with rhBMP-2, using diffusion chambers, into athymic mice induced ALP-positive cells in the chambers, but neither cartilage nor bone was observed. These results suggest that rhBMP-2 is a potent stimulator of osteoblast differentiation and bone formation in human cells. PMID- 9737345 TI - Calcitonin responsiveness and receptor expression in porcine and murine osteoclasts: a comparative study. AB - The presence of the calcitonin (CT) receptor is a distinguishing characteristic of osteoclasts; however, species variability exists with respect to functional responsiveness to CT. In the present study, CT responsiveness and temporal expression of the CT receptor in differentiating cultures of porcine osteoclasts was examined and compared to murine osteoclasts. In vitro porcine osteoclast differentiation was evaluated using bone marrow cultures from neonatal pigs. Murine osteoclast differentiation was studied using cocultures of murine bone marrow and BALC cells, a calvarial-derived cell line. In the presence of 1,25 (OH)2D3, a time-dependent increase in osteoclast differentiation was observed in porcine and murine cultures. Salmon CT (sCT) and porcine CT (pCT) inhibited 1,25 (OH)2D3-stimulated porcine osteoclast differentiation at 10(-8) and 10(-7) mol/L (60% with 10(-7) mol/L sCT and 85% inhibition with 10(-7) mol/L pCT). Treatment of murine cocultures with sCT (10(-17)-10(-7) mol/L) resulted in a concentration dependent decrease in osteoclast differentiation with a maximal inhibition of 70%. Osteoclast differentiation was inhibited in a concentration-dependent manner by recombinant human transforming growth factor-beta1 (rhTGF-beta1) in both species. The effects of CT on resorption lacunae formation were determined by culturing in vitro generated porcine or murine osteoclasts on bovine cortical bone slices for 18 h in the presence or absence of CT. With both porcine and murine osteoclasts, a concentration-dependent decrease in resorption lacunae formation was observed between 10(-13) and 10(-7) mol/L sCT with the highest concentrations completely abolishing resorption. However, pCT only inhibited porcine osteoclastic resorption at 10(-7) mol/L. CT receptor messenger ribonucleic acid (mRNA) expression was determined at different time points during in vitro osteoclast differentiation. In porcine cultures, expression of CT receptor mRNA correlated with the presence of osteoclasts. In murine cocultures, mRNA for the CT receptor was observed at each time point examined and was independent of the presence of multinucleated osteoclasts. Thus, porcine and murine differentiating osteoclast cultures express CT receptor mRNA; however, receptor expression correlates with osteoclast formation only in the porcine cultures. In summary, porcine and murine osteoclasts express CT receptor mRNA and functional responsiveness to CT. These findings suggest that the effects of sCT on osteoclast resorption are similar in murine and porcine cells, but that sCT is a less potent inhibitor of porcine than murine osteoclast differentiation. PMID- 9737346 TI - Rostrum of a toothed whale: ultrastructural study of a very dense bone. AB - The rostral bones of the toothed whale, Mesoplodon densirostris, consist mainly of hypermineralized secondary osteons and have yielded among the highest values for density (2.6 g/cm3) and mineral content (86.7%) yet reported for any bone. Scanning and transmission electron microscopy show parallel rods of mineral oriented along the length of the rostrum. These consist of platey crystals of carbonated hydroxyapatite, which, judging from electron diffraction, are extremely well and coherently aligned. The collagen component of the rostral bone consists largely of very thin fibrils aligned in longitudinal register to form tubular networks. The collagen fibrils are also aligned with the lengths of the mineral rods, which are apparently accommodated in the tubular spaces of the collagenous network. This peculiar ultrastructure clearly differs from the densely packed mineralized fibrils commonly observed in vertebrate lamellar osseous tissues, although histological examination has indicated some vestiges of "normal" primary bone surrounding the secondary osteons. Thus, the bone tissue in the rostrum is characterized by a remarkably sparse collagenous component. This ultrastructure can explain the high density, stiffness, and brittleness of the rostrum that have been observed. It also raises interesting questions about possible modes of crystal growth during ongoing mineralization in normal bone, and may have some relevance in the mechanical behavior of dense bones in pathological conditions. PMID- 9737347 TI - Prostaglandin E2 increases bone strength in intact rats and in ovariectomized rats with established osteopenia. AB - It is well documented that prostaglandin E2 (PGE2) has the ability to stimulate bone formation, improve bone structure, and increase bone mass in intact or osteopenic rat models. However, the effects of PGE2 on the mechanical properties of bone have not been investigated previously. The purpose of our study was to determine the effects of PGE2 on the mechanical strength of bones in rapidly growing, adult, and ovariectomized rat models. In study I, PGE2 at 3 mg/kg per day, or vehicle, was given by daily subcutaneous injections for 30 days to rapidly growing (3-month-old) intact male rats. Compared with controls, PGE2 significantly increased initial maximal load and stiffness of cancellous bone at the distal femoral metaphysis (DFM) as determined by an indentation test. As determined by a compression test, rats treated with PGE2 showed a significant increase in maximal load, and a nonsignificant increase in stiffness in the fifth lumbar vertebral body (L5) when compared with controls. In study II, PGE2 at 3 mg/kg per day, or vehicle, was given by daily subcutaneous injection for 30 days to mature (10-month-old) intact male rats. PGE2 treatment significantly increased initial maximal load and stiffness of the DFM and L5. PGE2 induced a significant increase in maximal load, but not stiffness, in the femoral neck (FN), as determined by a cantilever compression test. There was an increase in maximal load in a three-point bending test at the femoral shaft (FS) although the increase did not achieve statistical significance. No change in stiffness in the FS was found after PGE2 treatment. In study III, 3-month-old female rats were sham-operated or ovariectomized (ovx) for 30 days. Thereafter, PGE, at 1 or 3 mg/kg, or vehicle, were given by daily subcutaneous injection to these rats for 30 days. After 30 and 60 days, ovx induced a significant decrease in initial maximal load and stiffness of cancellous bone at the DFM as compared with sham controls. In ovx rats with established osteopenia, PGE2 at 1 mg/kg per day nonsignificantly increased the initial maximal load and stiffness, whereas, at 3 mg/kg per day, PGE2 completely restored the initial maximal load and stiffness of DFM to sham control levels. Similarly, maximal load and stiffness of L5 decreased significantly in ovx rats compared with sham controls at 30 days postsurgery. PGE2 at 1 mg/kg per day partially restored the maximal load, whereas, at 3 mg/kg per day, it completely restored the maximal load and stiffness of L5 in the established osteopenia, ovx rats. At the FS, PGE2 at 3 mg/kg per day nonsignificantly increased maximal load (+11%) and significantly increased stiffness (+25%) compared with ovx controls. Neither ovx nor PGE2 treatment caused a significant change in the maximal load and stiffness of the FN in this study. These results reveal that PGE2 significantly increased the mechanical strength at various skeletal sites in rapidly growing and mature male rats, although the increase in femoral shafts was not statistically different. Furthermore, PGE2 completely restored mechanical strength to the cancellous bone in ovx rats with established osteopenia. PMID- 9737348 TI - Disparate effects of mild, moderate, and severe secondary hyperparathyroidism on cancellous and cortical bone in rats with chronic renal insufficiency. AB - The subtotally nephrectomized rat has often been used to investigate the etiology and treatment of secondary hyperparathyroidism (secondaryHPT), but it has been used less frequently to study the effects of secondaryHPT on bone. The recent development of a reliable and specific rat parathyroid hormone (PTH) immunoradiometric assay has provided an opportunity for a thorough investigation of the relationship between circulating, biologically active PTH, and the skeletal abnormalities that occur in chronic renal insufficiency (CRI). Rats were 5/6 nephrectomized (Nx) or sham operated and fed diets with varying levels of Ca and P for 12-14 weeks to induce differing magnitudes of secondaryHPT. Parathyroid gland volume increased by 80%-90% in 5/6 Nx rats in the mild and moderate secondaryHPT groups (2.3- and 7.7-fold higher PTH levels, respectively) and by 3.3-fold in the severe secondaryHPT group (12-fold increase in PTH). The increases in gland volume were caused primarily by cell hyperplasia. Mild secondaryHPT resulted in a 12% decrease in bone mineral density (BMD) across the entire femur, increased osteoclast numbers (N.Oc), unchanged osteoblast numbers (N.Ob), and decreased cancellous bone volume (Cn.BV) in the tibial metaphysis but, apart from increased marrow area, no major changes in cortical bone at the tibio-fibular junction. Moderate secondaryHPT was associated with no changes in femoral BMD, or in tibial Cn.BV, but N.Ob and bone formation rate (BFR) were markedly elevated. Increased periosteal, intracortical, and endocortical BFR and turnover were evident, and contributed to increased cortical porosity (Ct.Po). The changes were exaggerated in the severe secondaryHPT group; BMD was lower in the proximal, but higher in the distal femur, and Cn.BV, N.Ob, N.Oc, and BFR were increased by six-, seven-, three-, and 30-fold, respectively. Endocortical BFR was elevated 31-fold and the extensive Ct.Po (10%) decreased bone strength. However, Ct.Po was not apparent until PTH levels exceeded 500 pg/mL. Thus, in rats with CRI of similar magnitude, progressive secondaryHPT is associated with dramatically different effects on bone. Mild secondaryHPT caused loss of cancellous and endocortical bone, and moderate secondaryHPT tended to maintain both types of osseous tissue, whereas PTH levels >500 pg/mL resulted in substantial cortical bone loss, but cancellous bone gain. PMID- 9737349 TI - Comparison of the response of pelvic and proximal tibial cancellous bone in rat to ovariectomy with estrogen replacement. AB - In this study, we found that the trabecular architecture of the rat pelvis has similarities to that of human iliac crest. Although we made no direct comparisons between the estrogen deficiency-induced rat osteopenia model and postmenopausal histomorphometry of iliac crest, we attempted to determine whether the rat pelvis might be appropriate to study changes in bone modeling and in situ changes in osteoblast protein expression. Three groups of young, sexually mature rats (12 weeks of age, each group comprising six animals) were either ovariectomized (ovx) and treated with 17beta-estradiol (ovx + E), vehicle (ovx), or sham-operated (sham). Histomorphometric variables were quantitated in the pelvis and compared with proximal tibial metaphysis in the three groups. Immunocytochemical localization of osteocalcin was also evaluated in the two skeletal sites. There was a greater reduction in bone volume of the proximal tibial metaphysis of ovx rats than in the pelvis of ovx rats when compared with sham-operated animals (p < 0.01), although bone formation rates were significantly higher at the pelvic site than tibial metaphysis (p < 0.01). The more rapid loss of bone between the tibia and pelvis may reflect differences in longitudinal growth in young rats, but the other intersite differences in bone remodeling consequent to ovx were at least as well demonstrated in the pelvic trabecular structure. Because ex vivo removal of the rat pelvis is simple, and provides a larger histomorphometric section with which to evaluate dynamic changes in metabolic bone disease, we suggest that this site may be useful in studies of osteopenia in the sexually mature female rat. Immunocytochemical demonstration of osteocalcin in trabecular surface osteoblasts was excellent in both sites. These results suggest that the rat pelvis is as accessible for histological study as the more conventional appendicular sites. When compared with the proximal tibial metaphysis, the rat pelvis (1) has a more homogeneous trabecular structure; (2) has more than twice as much trabecular bone area to sample; (3) has no open epiphyseal growth cartilages; (4) loses trabecular bone half as rapidly after ovx; (5) displays a greater increase in bone turnover after ovx; and (6) is the same anatomic site that is sampled in humans. We have also shown that the pelvis is a suitable site to demonstrate immunocytochemistry for osteoblast-derived proteins. PMID- 9737350 TI - Intracortical remodeling in adult rat long bones after fatigue loading. AB - Intracortical remodeling in the adult skeleton removes and replaces areas of compact bone that have sustained microdamage. Although studies have been performed in animal species in which there is an existing baseline of remodeling activity, laboratory rodents have been considered to have limited suitability as models for cortical bone turnover processes because of a lack of haversian remodeling activity. Supraphysiological cyclic axial loading of the ulna in vivo was used to induce bending with consequent fatigue and microdamage. Right ulnae of adult Sprague-Dawley rats were fatigue-loaded to a prefailure stopping point of 30% decrease in ulnae whole bone stiffness. Ten days after the first loading, left ulnae were fatigued in the same way. Ulnae were harvested immediately to allow comparison of the immediate response of the left ulna to the fatigue loads, and the biological response of the right leg to the fatigue challenge. Histomorphometry and confocal microscopy of basic fuchsin-stained bone sections were used to assess intracortical remodeling activity, microdamage, and osteocyte integrity. Bone microdamage (linear microcracks, as well as patches of diffuse basic fuchsin staining within the cortex) occurred in fatigue-loaded ulnar diaphyses. Ten days after fatigue loading, intracortical resorption was activated in ulnar cortices. Intracortical resorption occurred in preferential association with linear-type microcracks, with microcrack number density reduced almost 40% by 10 days after fatigue. Resorption spaces were also consistently observed within areas of the cortex in which no bone matrix damage could be detected. Confocal microscopy studies showed alterations of osteocyte and canalicular integrity around these resorption spaces. These studies reveal that: (1) rat bone undergoes intracortical remodeling in response to high levels of cyclic strain, which induce microdamage in the cortex; and (2) intracortical resorption is associated both with bone microdamage and with regions of altered osteocyte integrity. From these studies, we conclude that rats can initiate haversian remodeling in long bones in response to fatigue, and that osteocyte death or damage may provide one of the stimuli for this process. PMID- 9737351 TI - Vertebral bone density evaluated by dual-energy X-ray absorptiometry and quantitative computed tomography in vitro. AB - Vertebral bone density is evaluated mainly by dual-energy X-ray absorptiometry (DXA) or quantitative computed tomography (QCT). Densitometry is used as an estimator of bone strength and forms the basis for choice of treatment. DXA expresses bone density in grams per square centimeter (area density) and QCT expresses bone density in milligrams per cubic centimeter (volumetric density). The aim of this study was to identify the differences between the two techniques, DXA and QCT, when applied to a group of female and male subjects over a wide age range. The data consisted of 221 lumbar vertebral bodies (L3 and L4) excised at autopsy. There were 90 females with a mean age of 65.6 (range 18-94) years and 131 males with a mean age of 62.0 (range 21-94) years. The vertebrae were scanned en bloc in demineralized water in Plexiglas containers with both DXA and QCT. DXA was performed using posteroanterior (PA) and lateral projection. QCT was performed in the center of each vertebra with 1 cm slice thickness. Both methods showed decreasing bone density with age. Lateral DXA showed a decrease in bone density with age from approximately 0.8 g/cm2 to approximately 0.4 g/cm2. QCT showed a decrease in bone density with age from approximately 180 mg/cm3 to approximately 30 mg/cm3. Lateral DXA bone mineral densities (BMD) were correlated with QCT densities in both females (r2 = 0.68, p < 0.00001) and males (r2 = 0.53, p < 0.00001), but females had constantly lower DXA BMDs than males at a given QCT density. QCT and width-adjusted midlateral DXA (g/cm3) were significantly correlated, with r2 = 0.64 (p < 0.00001) for females and r2 = 0.61 (p < 0.00001) for males. In conclusion, age- and gender-related differences in human vertebral bone density were shown to be dependent on the scanning method used. DXA bone mineral content (BMC) and BMD showed that females had lower values than males at all ages. When the "volumetric" DXA measurements and QCT were used, the females had the highest densities in the younger decades and males had the highest densities in the oldest decades. Finally, the area density (BMD) measured by DXA was lower in females than in males with identical QCT volumetric bone densities. PMID- 9737352 TI - Menatetrenone ameliorates osteopenia in disuse-affected limbs of vitamin D- and K deficient stroke patients. AB - Significant reduction in bone mineral density (BMD) occurs in stroke patients on the hemiplegic and contralateral sides, correlating with the degree of paralysis and vitamin D and K deficiency due to malnutrition, and increasing the risk of hip fracture. We evaluated the efficacy of vitamin K2 (menatetrenone: menaquinone 4; MK-4) in maintaining BMD by comparing serum biochemical indices of bone metabolism between treated and untreated patients. In a random and prospective study, of 108 hemiplegic patients following stroke, 54 received 45 mg menatetrenone daily (MK-4 group, n = 54) for 12 months, and the remaining 54 (untreatment group) did not. Nine patients excluded from the study. The BMD in the second metacarpals and serum indices of bone metabolism were determined. BMD on the hemiplegic side increased by 4.3% in the MK-4 group and decreased by 4.7% in the untreated group (p < 0.0001), while BMD on the intact side decreased by 0.9% in the MK-4 group and by 2.7% in the untreated group (p < 0.0001). At baseline, patients of both groups showed vitamin D and K1 deficiencies, high serum levels of ionized calcium, pyridinoline cross-linked carboxyterminal telopeptide of type I collagen (ICTP), and low levels of parathyroid hormones (PTH) and bone Gla proteins (BGP), indicating that immobilization-induced hypercalcemia inhibits renal synthesis of 1, 25-dihydroxyvitamin D (1, 25-[OH]2D) and compensatory PTH secretion. Both vitamins K1 and K2 increased by 97.6% and 666.9%, respectively, in the MK-4 group. Correspondingly, a significant increase in BGP and decreases in both ICTP and calcium were observed in the MK-4 group, in association with a simultaneous increase in both PTH and 1, 25-[OH]2D. One patient in the untreated group suffered from a hip fracture, compared with none in the MK-4 group. The treatment with MK-4 can increase the BMD of disused and vitamin D- and K-deficient hemiplegic bone by increasing the vitamin K concentration, and it also can decrease calcium levels through inhibition of bone resorption, resulting in an increase in 1, 25-[OH]2D concentration. PMID- 9737353 TI - Effects of short-term treatment with prednisolone and calcitriol on bone and mineral metabolism in normal men. AB - To study the effects of treatment with glucocorticoid and calcitriol on biochemical markers of calcium and bone metabolism, 48 normal male volunteers (aged 21-54 years) were randomized to treatment for 7 days with either (A) prednisolone, 10 mg twice daily, (B) prednisolone, 10 mg twice daily, and calcitriol, 1 microg twice daily, (C) calcitriol 1 mg twice daily, or (D) placebo. The study period was 28 days. Renal calcium excretion increased (mean maximal increase +44.7%, p < 0.01) as well as serum parathyroid hormone (PTH) (max. +18.5%, p < 0.01) during prednisolone treatment. Concomitant treatment with calcitriol or calcitriol alone equally enhanced renal calcium excretion (max. +185.2%, p < 0.001) and decreased serum PTH (max. -43.1%, p < 0.001). Prednisolone administration was followed by prompt declines in markers of bone formation [serum osteocalcin (max. -34.7%, p < 0.001) and serum procollagen type I C-terminal propeptide (PICP) (max. -25.9%, p < 0.05)], whereas serum bone alkaline phosphatase (bone AP) remained unchanged. Calcitriol in combination with prednisolone attenuated the decrease in PICP (max. -8.9%, not significant), but it had no effect on osteocalcin (max. -40.1%, p < 0.001), and decreased bone AP (max. -22.2%, p < 0.05). Calcitriol alone increased osteocalcin (max. +37.8%, p < 0.03) and PICP (max. +26.0%, p < 0.05). Among markers of bone degradation, prednisolone suppressed serum C-terminal telopeptide of type I collagen (ICTP) (max. -28.4%, p < 0.001), but not the fasting renal excretion of hydroxyproline (OHP) and collagen type I N-terminal telopeptide (NTx). Calcitriol partially antagonized the decrease in ICTP (max. -17.2%, p < 0.001). Calcitriol alone had no effect on resorptive markers. Extraosseous matrix synthesis was suppressed by prednisolone evaluated by serum procollagen type III N-terminal propeptide (max. 30.8%, p < 0.001) and was not affected by concomitant treatment with calcitriol or calcitriol alone. In conclusion, short-term administration of prednisolone to healthy men leads to fast and protracted suppression of biochemical markers of bone formation and extraosseous connective tissue metabolism. The effect on bone was partially antagonized by simultaneous calcitriol treatment, and points toward a potential role of calcitriol in the prevention of steroid induced osteoporosis. PMID- 9737354 TI - Influence of microdamage on fracture toughness of the human femur and tibia. AB - The relationship between microdamage accumulation and bone fragility is not well understood. Previous work has demonstrated a positive relationship between microdamage and age in human cortical bone. Prior investigations have also demonstrated that fracture toughness decreases with age in the same bone. Based on these findings, the objective of this study was to test the hypothesis that a decrease in fracture toughness can be attributed to an increase in microdamage density. Microdamage parameters (density, surface density, and average crack length) were measured from bone taken from the shaft of the human femur and tibia and correlated with results from fracture toughness tests of the same bone. Results indicated that there was a weak but significant inverse relationship between fracture toughness and microdamage parameters for tension loading of the femur. These findings suggest that in vivo microdamage observable at the transmitted light level (100x) plays a secondary role to other contributory factors to decreased fracture toughness with age. Results also indicate that this relationship depends on bone ductility that apparently differs between the femur and the tibia. This study, in addition to prior investigations, suggests that crack size (microscopic vs. submicroscopic) and crack origin or type (in vivo vs. stress induced de novo) may influence the relationship between microdamage and bone toughness. PMID- 9737355 TI - Modification of the in vivo four-point loading model for studying mechanically induced bone adaptation. AB - We modified the noninvasive, in vivo technique for strain application in the tibiae of rats (Turner et al., Bone 12:73-79, 1991). The original model applies four-point bending to right tibiae via an open-loop, stepper-motor-driven spring linkage. Depending on the magnitude of applied load, the model produces new bone formation at periosteal (Ps) or endocortical surfaces (Ec.S). Due to the spring linkage, however, the range of frequencies at which loads can be applied is limited. The modified system replaces this design with an electromagnetic vibrator. A load transducer in series with the loading points allows calibration, the loaders' position to be adjusted, and cyclic loading completed under load control as a closed servo-loop. Two experiments were conducted to validate the modified system: (1) a strain gauge was applied to the lateral surface of the right tibia of 5 adult female rats and strains measured at applied loads from 10 to 60 N; and (2) the bone formation response was determined in 28 adult female Sprague-Dawley rats. Loading was applied as a haversine wave with a frequency of 2 Hz for 18 sec, every second day for 10 days. Peak bending loads were applied at 33, 40, 52, and 64 N, and a sham-loading group was included at 64 N. Strains in the tibiae were linear between 10 and 60 N, and the average peak strain at the Ps.S at 60 N was 2664 +/- 250 microstrain, consistent with the results of Turner's group. Lamellar bone formation was stimulated at the Ec.S by applied bending, but not by sham loading. Bending strains above a loading threshold of 40 N increased Ec lamellar bone formation rate, bone forming surface, and mineral apposition rate with a dose response similar to that reported by Turner et al. (J Bone Miner Res 9:87-97, 1994). We conclude that the modified loading system offers precision for applied loads of between 0 and 70 N, versatility in the selection of loading rates up to 20 Hz, and a reproducible bone formation response in the rat tibia. Adjustment of the loader also enables study of mechanical usage in murine tibia, an advantage with respect to the increasing variety of transgenic strains available in bone and mineral research. PMID- 9737356 TI - Ectopic expression of bone sialoprotein in human thyroid cancer. AB - Bone sialoprotein (BSP) is a small, highly posttranslationally modified integrin binding protein found in the mineral compartment of developing bone. The recent discovery that BSP can be detected in a variety of human cancers, particularly those that metastasize preferentially to the skeleton, shed light on potential new biological functions for this protein. The demonstration of a positive association between BSP expression in primary breast tumors and the development of bone metastases suggests that this glycoprotein could play a role in the selective implantation of breast cancer cells in bone. BSP is also expressed in most lung and prostate cancers as well as in multiple myeloma, three other osteotropic malignancies. Because thyroid carcinoma also metastasizes preferentially to the skeleton, we decided to look at the expression of BSP in a collection of 145 thyroid malignant lesions including 24 follicular thyroid carcinomas (FTCs), 55 papillary thyroid carcinomas (PTCs), 19 medullary thyroid carcinomas (MTCs), 23 anaplastic carcinomas (ACs), and 24 poorly differentiated carcinomas (PDCs). BSP expression was evaluated by immunoperoxidase technique using two specific polyclonal antibodies. Most of the thyroid carcinomas (72%) examined expressed high levels of BSP. Expression of BSP was significantly lower in FTCs and MTCs compared with PDCs, which are more aggressive (p = 0.0009 and 0.0003, respectively). Our study demonstrates for the first time that ectopic BSP expression is a common feature of thyroid cancer. The prognostic value of BSP detection in thyroid adenocarcinoma and the potential role of BSP in the propension of this type of cancer to metastasize to bone are currently under investigation. PMID- 9737357 TI - Comparative mapping of cloned human and murine antithyroglobulin antibodies: recognition by human antibodies of an immunodominant region. AB - Antibodies (Ab) to thyroglobulin (Tg) are common in patients with autoimmune thyroid diseases, but it is currently unclear how Tg Ab are involved in the pathology of autoimmune thyroid disease. We have previously reported the isolation of immunoglobulin G (IgG)kappa and IgGlambda Fab from phage display combinatorial libraries from the cervical lymph node of a single Hashimoto's thyroiditis patient with a high anti-Tg titer. Sequence analysis of these Fab indicated a restricted heavy chain usage with a nonrestricted light chain usage, with Fab inhibiting the binding of patient Tg Ab by between 39% and 79%. Comparative mapping of nine each of these IgGkappa and IgGlambda Fab, and the patient serum from whom the Fab were derived, is described here, using a panel of 10 murine monoclonal antibodies (Mab) to human thyroglobulin (hTg). The Fab interacted principally with mAb defining the overlapping antigenic domains I and IV, previously characterized as the region recognized by the majority of patient serum Tg Ab. Tg Ab from serum of the patient from whom the Fab were derived were also directed at this region, suggesting that the Fab are representative of the Tg Ab present in this patient. PMID- 9737358 TI - Thyroid nodules in Graves' disease: classification, characterization, and response to treatment. AB - Thyroid nodules in patients with Graves' disease are common and raise concern about coexistent thyroid malignancy. Alternative etiologies for such nodules are more frequent, and separation from thyroid malignancy is important for rational management. To characterize the types of thyroid nodules present in patients with Graves' disease, evaluate the response of these nodules to treatment, and stratify the risk of thyroid malignancy, we report on a retrospective single center study in an ambulatory setting of 468 Graves' patients ages (12-75) followed for 1-31 years (mean = 5.1) treated with radioiodine (n = 345), near total thyroidectomy (n = 19), thionamide antithyroid drugs (n = 88) or observation (n = 18). Sixty patients (12.8% of the total) had nodules and were classified as: (1) Graves' disease with a solitary hypofunctional nodule (n = 27, 5.8%); (2) Graves' disease with multiple nodules (n = 21, 4.5%); (3) Graves' disease with autonomous nodule (n = 4, 1%); or (4) patchy Graves' disease (n = 8, 1.7%). Six patients (1.3% of total or 10% of nodule patients) had cancer: 5 in group 1 and 1 in group 4. Based on the response to therapy or surgical and fine needle aspirate pathology, the remaining patients demonstrated pseudo-nodules of autoimmune thyroid disease, autonomous nodules of Marine-Lenhart syndrome, colloid goiter, hyperplastic adenomatous disease, and Hashitoxicosis. In conclusion, Graves' patients present with or may develop nodules commonly (12.6%) and the majority of these are benign expressions of autoimmune changes and coexistent nodular goiter. Thyroid cancer occurs in 10% of all nodules, 19% of palpable solitary cold nodules and 1.3% of the total patients. If the fine-needle aspiration biopsy (FNAB) cytology is benign, it is reasonable to use nonsurgical therapy. Any single cold nodule that remains or develops after treatment needs careful re-examination due to the high risk of malignancy. PMID- 9737359 TI - Quality of life aspects and costs in treatment of Graves' hyperthyroidism with antithyroid drugs, surgery, or radioiodine: results from a prospective, randomized study. AB - The patients' views and costs of three different forms of treatment for Graves' hyperthyroidism were investigated. The study comprises 174 patients with Graves' hyperthyroidism who were stratified into two age groups: 20 to 34 years and 35 to 55 years. The younger group was randomly assigned to treatment with antithyroid drug plus thyroxine for 18 months or subtotal thyroidectomy, and in the older group iodine-131 was added as a third alternative. The patients' views of their therapy were based on a questionnaire formulated to identify possible differences between the three treatment forms. The costs were assessed by analyzing the official hospital reimbursement system for both outpatient and inpatient costs for a period of 2 years from the day of randomization. The results show that no significant differences in opinion were found between the five treatment groups with regard to any of the questions. Furthermore, only 10% of the patients expressed slight and 3% major hesitation to recommend the treatment form received to a friend with similar disease. Twenty percent of the patients with endocrine ophthalmopathy reported the eye problems to be much more troublesome and 14% somewhat more troublesome than the thyroid problems. The cost proportion between the medical and surgical treatment in the young group was 1:2.5 (1 = 1126 United States dollars [USD]) before and 1:1.3 (1 = 2284 USD) after inclusion of the relapse costs. The proportion between the medical, surgical, and iodine-131 treatment in the older group was 1:2.5:1.6 (1 = 1164 USD) before and 1:1.6:1.4 (1 = 1972 USD) after inclusion of the relapse costs. PMID- 9737360 TI - Comparison of the results of diagnosis and treatment between solid and cystic well-differentiated thyroid carcinomas. AB - We assessed the accuracy of ultrasonography and fine-needle aspiration cytology (FNAC) in diagnosing cystic thyroid cancer and compared the results with solid thyroid cancer patients. We also compared the results of treatment between these patient groups. We retrospectively reviewed 1013 thyroid cancer patients who received treatment at Chang Gung Memorial Hospital. For this study, 910 cases of papillary or follicular thyroid carcinomas were considered eligible. Of these, 682 patients received preoperative high-resolution ultrasonographic and FNAC examinations. The nodules of 583 (85.5%) patients were diagnosed as solid masses, 80 (11.7%) as mixed masses, and 19 (2.8%) as cystic masses. Of the 19 patients with cystic thyroid carcinoma, only 4 papillary thyroid carcinomas were diagnosed by ultrasonography with FNAC as malignant before operation. Six patients presented as occult thyroid carcinomas with the tumor size less than 1 cm. Despite the low rate of accurate diagnosis for the cystic malignancy, clinical staging and the survival rates were not statistically different when they were compared with the other groups. In conclusion, low diagnostic rates were observed in well-differentiated thyroid cancer with prospective ultrasound-guided FNA when lesions were cystic or in mixed lesions. If the solid portion of the cystic masses is aspirated under ultrasound-guided FNA and cytology is performed after the centrifugation of the aspirated fluid, diagnostic accuracy may be improved. PMID- 9737361 TI - Effects of pharmacological fiber supplements on levothyroxine absorption. AB - To determine the effect of pharmacological fiber supplements, we measured levothyroxine (LT4) absorption without and with simultaneous ingestion of either calcium polycarbophil or psyllium hydrophilic mucilloid. Serum thyroxine (T4) levels in 8 volunteers were measured following ingestion of 600 microg of LT4 on 3 separate occasions at 4-week intervals: (1) LT4 alone; (2) LT4 together with 1000 mg polycarbophil; and (3) LT4 together with 3.4 g psyllium. The amount of absorbed LT4 was calculated as the incremental rise in serum T4 level during the first 6 hours multiplied by the volume of distribution for the hormone, and expressed as a percentage of the dose administered. Absorption of LT4 alone averaged 89% (95% confidence interval [CI]: 75%-104%), occurring at a median of 180 minutes. After simultaneous ingestion of calcium polycarbophil, LT4 absorption was 86% (95% CI: 74%-97%), occurring at 180 minutes. With simultaneous ingestion of psyllium and LT4, the absorption was 80% (95% CI: 64%-95%), occurring at 240 minutes. In summary, neither calcium polycarbophil nor psyllium hydrophilic mucilloid are likely to cause malabsorption of LT4 that could be detected by these methods. PMID- 9737362 TI - Hyperthyroidism due to lymphoma involving the thyroid gland in a patient with acquired immunodeficiency syndrome: case report and review of the literature. AB - A 31-year old woman with acquired immunodeficiency syndrome (AIDS) and a history of lymphoma presented with a 2-week history of severe hyperthyroid symptoms and new-onset neck swelling. On physical examination, she was found to be clinically hyperthyroid, with a markedly enlarged, diffuse, tender goiter. Thyroid function testing confirmed hyperthyroidism. The patient had a rapidly deteriorating clinical course and died within days of her presentation. At autopsy, near complete replacement of the thyroid gland with anaplastic large cell lymphoma was found, without coexisting infectious or autoimmune processes in the gland. This is the first case report of a patient with AIDS developing symptomatic thyroid involvement by lymphoma, and one of only a few case reports of hyperthyroidism associated with lymphoma in general. PMID- 9737364 TI - Relapse of Graves' disease after subacute thyroiditis. AB - We report the case of a 46-year-old man whose Graves' disease relapsed 5 years after its first appearance. At the time of relapse thyrotropin (TSH) receptor antibodies were very high, as was radioactive uptake in the left lobe of the thyroid. After thyroidectomy, histological analysis of the specimen showed evidence of treated Graves' disease, and there were signs of subacute thyroiditis in the right lobe. The clinical and immunologic characteristics of this observation are discussed as well as the hypothesis explaining the succession of an inflammatory and autoimmune thyroid disease. PMID- 9737363 TI - Reversible pituitary enlargement in the syndrome of resistance to thyroid hormone. AB - We report pituitary enlargement after radioiodine ablation in a patient with elevated thyroid hormones and features of hyperthyroidism. Serum thyrotropin (TSH) levels were elevated despite normal circulating thyroid hormones, suggesting inappropriate TSH secretion associated either with a TSH secreting pituitary adenoma or resistance to thyroid hormone (RTH). Normal serum glycoprotein alpha-subunit levels and a preserved TSH response to thyrotropin releasing hormone (TRH) favored RTH and this diagnosis was confirmed by showing the patient to be heterozygous for a missense mutation (R438H) in the thyroid hormone beta receptor (TRbeta) gene. Thyroxine replacement in supraphysiological doses were required to normalize TSH levels and resulted in regression of the pituitary enlargement, suggesting hyperplasia rather than coincident tumor. This case illustrates the need to avoid thyroid ablation in RTH patients and the importance of supraphysiological thyroxine replacement to prevent pituitary hyperplasia. PMID- 9737365 TI - Effects of cytokines on expression of thyrotropin receptor mRNA in rat preadipocytes. AB - Cultured rat preadipocytes express thyrotropin receptor (TSHR) during their differentiation. To evaluate the effects of inflammatory cytokines on the expression of TSHR in cultured rat preadipocytes, we cultured those cells in the presence of recombinant human tumor necrosis factor (rhTNF)-alpha, recombinant human interferon (rhIFN)-gamma, and human transforming growth factor (hTGF) beta1. The effects on the level of TSHR mRNA and signal transduction were evaluated. Addition to the medium of 1 ng/mL TNF-alpha, 1 ng/mL rhIFN-gamma, and 1 ng/mL hTGF-beta1 during the differentiation of rat preadipocytes inhibited the expression of TSHR mRNA. The decrease in TSHR mRNA was accompanied by a decrease in TSH-stimulated cyclic adenosine monophosphate (cAMP) production. Histochemical analysis showed that these cytokines inhibited the morphological differentiation of the cells. These cytokines also decreased the expression of mRNA for such fat specific proteins as lipoprotein lipase and aP2. Results indicate that the loss of expression and function of the TSHR is closely related to the inhibition of differentiation. This confirms the close relation between the expression of the TSHR and the differentiation of the rat preadipocytes. PMID- 9737366 TI - Development and characterization of monoclonal antibodies specific for the murine thyrotropin receptor. AB - We have characterized 10 monoclonal antibodies (Mabs) to recombinant murine thyrotropin receptor extracellular domain (mTSHR-ecd). Affinity purified mTSHR ecd (amino acids 22-415), expressed in a baculovirus-insect cell system, was refolded in vitro and used to hyperimmunize female Balb/c mice. Spleens were removed 10 days after a final boost of 25 microg mTSHR-ecd intraperitoneally and intravenously, and the cells were fused to SP-2 cells and cloned. Hybridoma supernatants were screened by enzyme-linked immunosorbent assay (ELISA) with folded mTSHR-ecd antigen. Ten of 18 higher affinity hybridomas were selected at random and ascites fluids prepared. Nine of the monoclonals were of IgG 1 isotype, and one was IgM. Five Mabs (M3, M4, M5, M6, and M9) inhibited the binding of 125I-TSH to functional hTSHR expressed on Chinese hamster ovary (CHO) cells, and four (M1, M3, M5, and M9) blocked the TSH-stimulated generation of cyclic adenosine monophosphate (cAMP), using the same cells. The remaining Mabs appeared to be neutral in their interaction with native TSHR. The Mabs were also compared for their reactivity to mTSHR-ecd under folding (ELISA) and unfolding (reducing sodium dodecyl sulfate-polyacrylamide gel electrophoresis [SDS-PAGE]) conditions. Most Mabs demonstrated reactivity to both conformational (folded) and linear (unfolded) forms of mTSHR-ecd, suggesting that they were generated primarily against linear epitopes although one Mab (M4) showed affinity for only folded antigen indicating a preference for a conformational epitope. Mapping the Mab epitopes using 26 overlapping peptides spanning the human (h)TSHR-ecd showed that 6 bound peptide 397-415, 1 bound peptide 352-371, and 1 peptide 22-41. These epitope mapped Mabs to the mTSHR-ecd, both receptor blocking and receptor neutral, will provide further insight into the structure-function of the TSHR ectodomain. PMID- 9737367 TI - Thyroid hormone receptor coactivators and corepressors. AB - In the absence of triiodothyronine (T3), thyroid hormone receptors (TRs) repress transcription of many genes; in the presence of T3, TRs activate transcription of those same genes. Both of these events are dependent on interactions between TRs and other nuclear proteins. TRs bind to specific DNA sequences, generally found in the 5' flanking regions of target genes. In the unliganded state, TRs interact with one of several corepressor proteins. These proteins, in turn, interact with a series of other proteins, which includes histone deacetylases. Histone deacetylation tightens chromatin structure, thus impairing access of critical transcription factors and thereby repressing transcription. In addition, corepressors may invoke mechanisms of gene repression independent of histone deacetylation. The binding of T3 causes a conformational change in the TR that results in release of the corepressor and recruitment of coactivator proteins. Several coactivator proteins appear to bind the ligand-occupied TR as a multiprotein complex. Opposite to corepressors, coactivators acetylate histones, thereby loosening chromatin structure and facilitating access of key transcription factors. Again, mechanisms independent of histone acetylation also may be involved. Overall, gene activation by T3 is a two-step process; removal of active repression, and induction of transcription to levels above the "neutral" state. PMID- 9737368 TI - Anaplastic thyroid carcinoma: behavior, biology, and therapeutic approaches. AB - Anaplastic thyroid carcinoma (ATC), although exceedingly rare, is the most aggressive solid tumor known. Early studies on the effects of different therapies may be biased by the inclusion of responsive "small cell" ATC patients, which are now known to be mostly lymphoma patients. Local control of disease with surgery and/or external beam radiotherapy (XRT) is of fundamental importance to enhance survival. Ultimately, nearly all ATC patients die from their disease, which is widely metastatic. Development of effective systemic chemotherapy agents would provide the best chance for long-term survival of patients. Early preliminary data suggest that paclitaxel may be helpful, although no agent has yet been identified to result in dramatic improvements in survival. Select patients may benefit from aggressive multimodal therapy, although it is important to provide appropriate palliative care when desired. PMID- 9737369 TI - What is the evidence of genetic factors in the etiology of Graves' disease? A brief review. AB - Graves' disease (GD) is generally thought of as a multifactorial disorder in which genetic susceptibility interacts with environmental and endogenous factors to cause disease. The importance of genetic factors is suggested by the clustering of GD within families and by a higher concordance rate for disease in monozygotic than dizygotic twins. This has, however, recently been shown to be less pronounced than previously thought. During the last decade, much effort has been put into characterization of the genetic background of GD. Until recently most studies have examined associations between GD and the human leukocyte antigen (HLA) region, but recent advances in molecular techniques have opened the way for whole-genome screening. A number of HLA and non-HLA candidate genes have been proposed, but despite several large investigations within multiplex families no major susceptibility genes have been identified. This brief review discusses relevant articles published from 1940 through 1997 regarding the influence of genetic factors in the etiology of GD. Ongoing studies will focus on whole genome screening in multiplex families as well as population based twin studies. However, the possibility of GD being a heterogeneous disease without a single well-defined genotype and phenotype should be left open. PMID- 9737371 TI - Karyology and tumorigenicity testing requirements: past, present and future. AB - At the present time, karyology and tumorigenicity are applied to primary, diploid, and continuous cell systems in an uneven and inappropriate manner, largely for historical reasons. It is a significant anomaly that such rigorous requirements are applied only to diploid cells when, of all three cell types, they represent the category with the least potential problems. During the 1992 presidential campaign in the U.S., a very direct and telling slogan was used to the advantage of Mr. Clinton: <>. That message was meant to have the effect of focusing on the real issue that was of concern to the electorate. The other topics were of secondary significance and tended to act as distractions from the central issue. In a very similar sense, one could say: <>. In other words, we should be focussing our time and attention on the characteristics of the biological product manufactured in a given cell system that has been well characterized rather than continue to belabour the issue of cell substrates. To a large degree, this has already been initiated with the recommendations of various groups already mentioned. However, current diploid cell quality control regulations stand out as a peculiar throwback to an earlier era. HDCs should be treated on a par with primary and continuous cells. There are three basic questions related to the routine use karyology that need to be addressed: (i) is the original rationale for requiring cytogenetic analysis of a diploid cell substrate still valid; (ii) is there a new rationale that would warrant its continued use; and (iii) if there is a continuing need for karyology, is it unique to diploid cell cultures or does it need to be extended to all types of cell systems? The original rationale was that karyology provided evidence of the normal character of WI-38 cells and therefore supported its acceptability as a cell substrate for vaccine production. Karyology essentially has remained as a legacy of the intense debate that led to the acceptance of WI-38 cells. There is no new information over the past 30 years to suggest that there is a new rationale for instituting chromosomal analysis of cell substrates. If there were, however, it is difficult to imagine why it should not be applied to all types of cell substrates (primary, diploid, and continuous). Taking all the above into consideration, there would seem to be no rationale for continuing to single out diploid cell cultures as the only cell type for which karyology is required on a routine basis. As stated above, the initial characterization of a new diploid cell line should include karyology. Like karyology, tumorigenicity testing was incorporated into the assessment of diploid cells in an attempt to persuade regulatory authorities that the cells were normal and acceptable. Again, after 30 years of testing, there has never been an instance of normal diploid fibroblasts generating a tumour in any in vitro or in vivo assays. The futility of continuing to do these tests is obvious. A description of the tumorigenic potential of a cell substrate should be an element in the characterization of a new cell line; but it has little if any value as a routine test. If history teaches us anything at all about risk, it is that we need to focus serious attention on contaminants rather than be diverted to remote theoretical issues that may be interesting to discuss and argue about, but which pale in the face of the potential impact on public health of viral and viral-like contaminants of biological products. One has only to recall the transmission of SV-40 from primary monkey kidney cells that were used to produce polio vaccine, or more recently the transmission of Creutzfeld-Jacob disease to recipients of human growth hormone derived from human pituitaries and of HIV to recipients of blood and blood derivatives. (ABSTRACT TRUNCATED) PMID- 9737372 TI - Cell line banking and authentication. PMID- 9737373 TI - Raw materials as a source of contamination in large-scale cell culture. AB - Large-scale cell culture operations for biotechnology products use millions of litres of complex media and gases as well as huge quantities of organic and inorganic raw materials. These raw materials must always be assumed to contain contamination by adventitious agents such as Minute Virus of Mice (MVM). Genentech has had experience in dealing with two such contaminations. Although the source of these contaminations was not positively identified, there was strong evidence to suggest that the virus entered through a raw material used in cell culture. Analytical methods that can be used to detect the presence of viruses can be used as an early warning system and as part of a strategy to devise barriers against such contamination. Methods such as a polymerase chain reaction (PCR) assay and a cell culture-based infectivity assay have been found to be efficacious in providing early detection of MVM infection. In any contamination, timely interactions with regulatory authorities are vital in minimizing delays in product manufacture. PMID- 9737374 TI - Zoonoses and haemorrhagic fever. AB - Virus zoonoses causing haemorrhagic fever have been recognized in three major families: Arenaviridae, Bunyaviridae and Filoviridae. All are negative-stranded RNA viruses, with genomes in two segments, three segments, or non-segmented, respectively. Acquisition of haemorrhagic fever in man generally requires close contact with a vertebrate vector species, usually rodents, for the arenaviruses and bunyaviruses. In the case of filoviruses, the vector is currently unknown, but these viruses may infect monkeys, and may contaminate cell cultures prepared from them. Both bunyavirus and arenavirus haemorrhagic fevers have arisen in humans following exposure to rodents, and in the case of Hantaan, a virus causing haemorrhagic fever with renal syndrome (HFRS), there have been numerous laboratory-acquired infections among animal care workers. As the technology to differentiate virus species has improved, it has become clear that there are numerous potentially hazardous viruses capable of causing HFRS or hantavirus pulmonary syndrome (HPS) within the feral rodent population. In many cases it would be desirable to introduce screening methods for such viruses before preparing cell cultures from these rodent or simian species that will be used to prepare biological products for human use. PMID- 9737375 TI - Overview of biological effects of addition of DNA molecules to cells. AB - Injected DNA proceeds with certain probabilities through the following steps: degradation by serum nucleases, adsorption to cells, uptake into cells, ligation to other DNA, mutation, expression of unintegrated DNA, integration, expression of integrated DNA, and activation of or inactivation of cellular genes. The maximal probability per DNA molecule of each of these steps is estimated based on experimental results in cell culture with transfection of DNA and with infection by retroviruses. A maximum cumulative probability of having a harmful effects is calculated to be less than 10(-16) to 10(-19) per DNA molecule from a cell without activated proto-oncogenes or active viral oncogenes. The most frequent harmful effects considered are inactivation of a tumour suppressor gene and activation of a proto-oncogene. Such inactivation and activation in a cell that could give rise to cancer would increase the age-standardized incidence of cancer by a small amount. The amount of increase would differ among individuals depending upon their genotypes and their environments. Thus, the magnitude of the increase will depend upon the frequency of more sensitive individuals. The probability of an increased incidence of cancer as a possible effect of the vaccination should be compared to the number of DNA molecules to be injected per person and to the protective effects of successful HBV vaccine. PMID- 9737376 TI - Risk potential of the chromosomal insertion of foreign DNA. PMID- 9737377 TI - DNA issues. AB - For many years the debate surrounding the potential danger of contaminating cellular DNA in biological products has focussed on the possibility that this DNA could give rise to tumours in the recipient host. Current knowledge of the nature and the fate of this DNA after in vivo administration allows us to conclude that this material represents a minimal risk to the patient. PMID- 9737378 TI - Residual host cell protein from continuous cell lines. Effect on the safety of protein pharmaceuticals. AB - The presence of residual host cell proteins in protein pharmaceuticals obtained from continuous mammalian cell lines has been a point of concern. Clinical experience with different biopharmaceuticals shows that residual protein in these highly purified products does not represent a danger to the health of the patients. PMID- 9737379 TI - An overview of the BSE epidemic in the UK. AB - The BSE epidemic in the UK is declining rapidly in response to control measures and is en route for eradication by the year 2001. BSE infectivity is restricted in clinically affected natural cases of BSE to central nervous tissues. BSE agent has been isolated from the intestine of experimentally orally-infected cattle from six to 18 months post-challenge. A small maternal risk factor for BSE has been identified but this will not stop the eradication of BSE. To protect the integrity of biological products from the risks associated with the unknown hazard of BSE for man it is recommended that only safe sources of animals and tissues are used. Inactivation procedures employed during processing will provide further guarantees. PMID- 9737380 TI - Transmission of spongiform encephalopathy through biological products. AB - The transmissible spongiform encephalopathies (TSE) are rare but fatal neurological diseases that exist in sporadic, familial, and environmentally acquired forms. Environmentally acquired disease has until now been mostly iatrogenic in origin, recognized as responsible for nearly 200 deaths during the past 20 years. The two most important causes have been contamination of cadaver derived human growth hormone and dura mater grafts, but a few instances related to contaminated neurosurgical instruments and corneal grafts have also occurred. Currently, a great deal of concern is being voiced about the potential risk of acquiring disease through blood or blood products, and although there is no supporting epidemiological evidence for such a danger, experiments are underway to define which blood components or plasma derivatives might pose the greatest problem. PMID- 9737381 TI - New variant Creutzfeldt-Jakob disease. AB - New variant Creutzfeldt-Jakob disease is a novel human spongiform encephalopathy with a consistent clinico-pathological phenotype. Epidemiological evidence indicates that this disease is occurring almost exclusively in the UK, where there has been an epidemic of spongiform encephalopathy in the cattle population. Current evidence strongly supports the hypothesis that there is a causal link between bovine spongiform encephalopathy and new variant Creutzfeldt-Jakob disease. PMID- 9737382 TI - Problems with BHK 21 cells. AB - The properties of baby hamster kidney (BHK 21) cells are modified by passage in suspension culture. The suspended cells differ from monolayer cells in the surface expression of some integrin chains involved in attachment of foot-and mouth disease virus (FMDV), in particular the progressive down-regulation of both alpha5 and alphaV integrin chains. This down-regulation is correlated with the loss of actin stress fibres. FMDV particles from these cells are unstable towards the aziridine used in inactivating the virus for vaccine production. Moreover, growth of virus in suspended cells can lead to the selection of antigenic variants which differ from those produced in monolayer cells. PMID- 9737383 TI - Experiences of virus, retrovirus and retrovirus-like particles in Chinese hamster ovary (CHO) and hybridoma cells used for production of protein therapeutics. AB - Garnick and coworkers indicated that they experienced two independent MVM outbreaks in a period where approximately 2000 fermentations were performed, hypothesizing that such events were rare but inevitable consequences of very large scale operations. In GIs experience over the last 12 years we have seen no incidence of MVM (or any other virus) in close to 3000 fermentations, albeit at lower volumes than produced at Genentech; GI has used 250-2500L bioreactors for manufacturing whereas Genentech have reported using 100-10,000L bioreactors. Nonetheless, volumes of complex media in the same range as used at Genentech have been used at GI with no observations of viral contamination events. The reason for this is not clear. However, GI's experience in combination with experience from sub-contract testing agencies who service the majority of the biotechnology industry may call the inevitability of an MVM outbreak into question. It would appear that very few adventitious viral contaminations of cell cultures have occurred in industry in the last decade. Interestingly, the frequency of contamination events appear to be lower in CHO cells than in hybridoma cells. It should be noted, however, that these conclusions are not statistically based and the scope of the above survey was somewhat limited. RVLPs are present in both CHO and hybridoma cells. The characteristics of both are compared in Table 4. C-type particles from hybridoma cells are more abundant as a rule than those from CHO cells. Although the majority of C-type particles produced by hybridoma cells appear to be non-infective (in S+L- assays), approximately one in a million particles are competent to replicate in S+L- cells. The evidence that C-type particles can replicate in human cells has proved difficult to reproduce consistently. It is likely that replication of xenotropic hybridoma C-type particles in human cells is inefficient or restricted to only a small number of specific cell lines. C-type RVLPs from CHO cells are produced less abundantly than those from hybridoma cells and are not competent to replicate due to a defective endonuclease gene. However, over the last two decades the use of hybridoma cells and products derived from these cells has not provided any evidence of transmission of these viruses to humans; in addition they can be readily removed or inactivated. Thus, neither agent would appear to constitute a significant risk to pharmaceutical products made from their respective host cells. Nonetheless, given the difference in relative safety profiles between RVLPs from CHO and hybridoma cells it is not unreasonable to propose that safety factors (clearance factors in removal/inactivation studies in excess of the reduction of virus loads to zero) required should be less for a CHO process than for a hybridoma process. PMID- 9737384 TI - Human diploid cell strains (HDCS) viral vaccines. AB - Since the development in 1961 by Hayflick and Moorhead of a human diploid cell strain (HDCS) culture system for isolating viruses, HDCS-derived human vaccines have been licensed worldwide for polio IPV and OPV (multiple strains), rabies (Pitman-Moore L503 3M strain); rubella (RA27/3 strain); measles (Edmonston-Zagreb strain); varicella-zoster (Oka strain); mumps (Rubini strain) and hepatitis A (HM 175, CR-326-F', and GBM strains). Many of these widely-used vaccines now have at least a 20-year record of safety after extensive, ongoing pharmacovigilance. Serious vaccine-associated events are rare, and appear to reflect the activity of the live viruses replicated in the HDCS or of the inactivated viruses or other proteins added during manufacture. There is no credible association of reactions to the HDCS substrate or a hypothetical contaminant derived from it. PMID- 9737385 TI - Chromosomal characterization of MRC-5 cell banks utilizing G-banding technique. AB - We have performed chromosomal monitoring on 18 MRC-5 cell banks using the G banding technique. A higher frequency of structural abnormalities and hyperdiploidy was detected with respect to the numerical values that were established with conventional staining techniques and reported in the 1979 Ad Hoc Committee guideline on karyology controls of human cell substrates. These numerical criteria have been adopted by international regulatory agencies to release human diploid cell banks (WI-38 and MRC-5) for vaccine manufacture. In the process of characterizing these 18 cell banks, a 7;12 translocation clone was detected in 7 cell banks at a frequency ranging from 0.2 to 5.6%. The presence of the t(7;12) appears to be biphasic. At low population doubling levels (PDLs) (< 30), t(7;12) is rarely observed. However, the incidence of t(7;12) increases and plateaus between PDL 40-50. A decrease in frequency is observed at higher PDLs. Before senescence of the cell bank, t(7;12) is not observed. Investigation of the growth characteristics of MRC-5 cells revealed that cell banks containing the translocation senesced at similar PDLs compared to MRC-5 cells with no detectable 7;12 translocation. In addition, MRC-5 cell banks containing the t(7;12) have successfully completed tumorigenicity testing in a nude mouse model. We conclude that MRC-5 cells obtained from either National Institute for Biological Standards and Control (NIBSC) or American Type Culture Collection (ATCC) contain a 7;12 translocation at a low frequency. This abnormality does not provide MRC-5 cell bank mass cultures with a growth advantage nor is it tumorigenic in nude mice. Furthermore, the presence of this clone and employment of the G-banding technique may be responsible for the increased observation of structural abnormalities in our laboratories. In view of this information, the cytogenetic criteria that were established in 1979 with conventional staining techniques are not appropriate for human diploid cell banks that are examined with more sensitive methodology. Since it has been recognized that MRC-5 and WI-38 cells are safe biological substrates, we recommend that MRC-5 and WI-38 cell banks should only be identified by using an appropriate identity test and should not require any chromosomal analysis before being used as a cell substrate for the manufacture of live virus vaccines. PMID- 9737386 TI - Experience with the Vero cell line. AB - The Vero cell line has been managed with the Cell Bank system to produce at the 142nd passage IPV, OPV and rabies vaccines since 1982 by Pasteur Merieux Serums & Vaccins (PMsv). The safety of the cell line was regularly validated at the Working Cell Bank (WCB) level according to the WHO and European Pharmacopoeia requirements for absence of bacteria, fungi, mycoplasma and viruses. A special emphasis was devoted to research on the absence of simian viruses (SV40, SIV, Retro-D virus and simian CMV). All these specific researches were negative. At a low level of passage, the Vero cells are not tumorigenic. Vaccines have been prepared in low passage level Vero cells and together with the excellent downstream purification have resulted in excellent safety as attested by pharmacovigilance of more than 100 million doses of IPV during 12 years, more than 20 million doses of rabies vaccine during 10 years and more than 1 billion of OPV during eight years. PMID- 9737387 TI - DNA in plasma of human blood for transfusion. PMID- 9737388 TI - Safety of biological products prepared from mammalian cell culture. Detecting viruses in cell banks. PMID- 9737389 TI - Safety of biological products prepared from mammalian cell culture. In-process testing for viral agents. PMID- 9737390 TI - Safety of biological products prepared from mammalian cell culture. Testing source materials. PMID- 9737391 TI - Safety of biological products prepared from mammalian cell culture. Bovine spongiform encephalopathy and other non-viral transmissible agents. PMID- 9737392 TI - Safety of biological products prepared from mammalian cell culture. Karyology. PMID- 9737393 TI - Safety of biological products prepared from mammalian cell culture. DNA. PMID- 9737394 TI - Safety of biological products prepared from mammalian cell culture. Host-cell proteins. PMID- 9737395 TI - WHO requirements for the use of animal cells as in vitro substrates for the production of biologicals (requirements for biological substances No 50). PMID- 9737396 TI - Quality of biotechnological products: viral safety evaluation of biotechnology products derived from cell lines of human or animal origin. ICH Harmonised Tripartite Guideline. PMID- 9737397 TI - Quality of biotechnological products: analysis of the expression construct in cell lines used for production of r-DNA derived protein products. ICH Harmonised Tripartite Guideline. PMID- 9737398 TI - Quality of biotechnological products: stability testing of biotechnological/biological products. Annex to the ICH Harmonised Tripartite Guideline for the Stability Testing of New Drug Substances and Products. PMID- 9737399 TI - Quality of biotechnological products: derivation and characterisation of cell substrates used for production of biotechnological/biological products. ICH Harmonised Tripartite Guideline. PMID- 9737400 TI - EMS Agenda for the Future: where we are...where we want to be. AB - During the past 30 years, emergency medical services (EMS) in the United States have experienced explosive growth. The American health care system is now transforming, providing an opportune time to examine what we have learned over the past three decades in order to create a vision for the future of EMS. Over the course of several months, a multidisciplinary steering committee collaborated with hundreds of EMS-interested individuals, organizations, and agencies to develop the EMS Agenda for the Future. Fourteen EMS attributes were identified as requiring continued development in order to realize the vision established within the Agenda. They are integration of health services, EMS research, legislation and regulation, system finance, human resources, medical direction, education systems, public education, prevention, public access, communication systems, clinical care, information systems, and evaluation. Discussion of these attributes provides important guidance for achieving a vision for the future of EMS that emphasizes its critical role in American health care. PMID- 9737401 TI - Outcomes of sudden cardiac arrest treated with defibrillation by emergency medical technicians (EMT-Ds) or paramedics in a two-tiered urban EMS system. AB - OBJECTIVE: Controversy exists as to the effectiveness of defibrillation by emergency medical technicians (EMT-Ds) in reducing mortality from cardiac arrest in two-tiered EMS systems. This study was performed to assess the impact of EMT Ds on outcome of sudden cardiac death in a small, urban, modified two-tiered EMS system. METHODS: This was a retrospective, unmatched case-control study comparing the outcomes of patients suffering sudden cardiac death treated by EMT-Ds with paramedic (EMT-P) backup with the outcomes of patients treated by EMT-Ps as first responders. Outcomes were defined as survival to the following endpoints: hospital admission, hospital discharge, and discharge with normal neurologic function (neurologic survival). Differences between groups were considered significant if p < or = 0.05 by Fisher's exact test or t-test. RESULTS: Three hundred twenty-two patients suffered out-of hospital sudden cardiac deaths over a three-year period and met study inclusion criteria. There were no significant differences in mean age, sex distribution, or incidence of ventricular fibrillation as the presenting rhythm between the groups. Rates of survival to admission, survival to discharge, and neurologic survival were 25.8%, 8.1%, and 5.6%, respectively. Corresponding survival rates for 46 patients treated first by EMT-Ds were 19.6%, 8.7%, and 4.3%. For 276 patients treated by EMT-Ps as first responders, the rates were 26.8%, 8.0%, and 5.8%. There were no significant differences in survival rates between the two response modes, despite a significantly shorter response interval for EMT-Ds (3.6 +/- 1.8 min, vs 4.6 +/- 2.0 min for EMT-Ps). There were likewise no significant differences in survival rates between the two response modes when only patients in ventricular fibrillation or ventricular tachycardia were considered. There were no significant differences in survival rates grouped by presenting rhythm, with the exception of 9.6% neurologic survival in witnessed ventricular fibrillation as compared with 0% in asystole. CONCLUSION: EMT defibrillation had no impact on outcome of sudden cardiac death in this small, urban, two-tiered EMS system. Survival rates were similar to those reported for other such systems. However, power to detect significant differences was low, and further study is indicated. Controlled multicenter trials are recommended. PMID- 9737402 TI - EMS-initiated refusal and alternative methods of transport. AB - OBJECTIVES: 1) To describe characteristics of patient transport protocols in those U.S. cities that sanction EMS-initiated refusal of transport; and 2) to describe the frequency and type of alternatives to emergency ambulance transport. METHODS: EMS systems in every one of the 200 largest cities in the United States were surveyed by telephone regarding EMS-initiated refusal policies, involvement of physicians in the decision-making process, and the presence or absence of alternatives to EMS transport. RESULTS: 100% of the target population responded to the telephone survey. Only 34 (17%) EMS systems have written protocols that allow EMS providers to refuse emergency ambulance transport for patients judged to have minor illness or injury after examination. Twenty-one (62%) of these EMS systems do not require on-line physician approval for EMS-initiated refusals. Seven (21%) EMS systems that allow refusal of transport also have a formalized alternative transport program in place. Nationwide, only 19 (10%) cities surveyed offer some type of alternative to ambulance transport, most commonly taxi and minivan. CONCLUSION: The authors report the first national survey of EMS initiated refusal practices. Few urban EMS systems have implemented this policy to decrease utilization by persons with low-acuity illness or injury. This may be related to the fact that few EMS systems currently have alternatives to emergency ambulance transport. PMID- 9737403 TI - The accuracy of medical records and police reports in determining motor vehicle crash characteristics. AB - OBJECTIVE: To determine the accuracy of police, emergency department, and ambulance records in describing motor vehicle crash (MVC) characteristics when compared with a crash investigation report (CIR). METHODS: This study was a retrospective record review. Sixty-three motor vehicle crash (MVC) patients transported to a university hospital emergency department via ambulance and also reported in a crash investigation record (CIR) during the period January 1993 to December 1995 comprised the study population. The crash characteristics analyzed were occupant position (OP), restraint use (RU), air bag deployment (AD), type of impact (TI), ejection (EJ), and external cause-of-injury code (EC). The accuracies of the police report (PR), the emergency department record (EDR), and the ambulance report (AR) for each patient were compared with the CIR by computing percent agreement, with 95% confidence intervals (95% CIs) for each variable and for each data source. RESULTS: Overall average agreement was 92.9% for PR, 89.7% for EDR, and 80.7% for AR. The overall average agreement for each variable was 98.9% for EJ, 92.1% for AD, 91.5% for OP, 90.5% for EC, 77.2% for RU, and 76.2% for TI. For all but one variable (RU), 95% CIs overlapped between data sources. CONCLUSIONS: The accuracy of data sources used to determine crash characteristics varies. Using a CIR as the standard, the PR was the most accurate. Inaccuracies occurred most frequently for RU and TI. Researchers and clinicians need to be aware of these inaccuracies. PMID- 9737404 TI - Out-of-hospital use of neuromuscular-blocking agents in the United States. AB - OBJECTIVE: A national survey was mailed to state EMS directors to assess the current status of use of neuromuscular-blocking agents (NMBAs) by EMS systems across the United States. METHODS: Initial and second mailings were completed in early 1996 and 1997, respectively. RESULTS: Replies were received from all 50 (100%) states following the second mailing. Twenty-nine (58%) states use NMBAs. Eleven (22%) of the 29 use paralytic drugs only in aeromedical programs. Of the 18 states that use NMBAs in ground-based EMS systems, 11 (22%) use paramedic ambulance staffing exclusively. Registered nurses (RNs) or RN-paramedic teams comprised the majority of the remaining states' staffing configurations. The first reported date of implementation of use of NMBAs was 1985, and there has been a steady trend of additional states launching paralytic drug use over the last 12 years. CONCLUSION: This trend suggests that use of paralytic drugs by paramedics is becoming standard of care in many out-of-hospital systems. PMID- 9737405 TI - Can EMS providers adequately assess trauma patients for cervical spinal injury? AB - OBJECTIVE: To determine whether EMS providers can accurately apply the clinical criteria for clearing cervical spines in trauma patients. METHODS: EMS providers completed a data form based on their initial assessments of all adult trauma patients for whom the mechanism of injury indicated immobilization. Data collected included the presence or absence of: neck pain/tenderness; altered mental status; history of loss of consciousness; drug/alcohol use; neurologic deficit; and other painful/distracting injury. After transport to the ED, emergency physicians (EPs) completed an identical data form based on their assessments. Immobilization was considered to be indicated if any one of the six criteria was present. The EPs and EMS providers were blinded to each other's assessments. Agreement between the EP and EMS assessments was analyzed using the kappa statistic. RESULTS: Five-hundred seventy-three patients were included in the study. The EP and EMS assessments matched in 78.7% (n = 451) of the cases. There were 44 (7.7%) patients for whom EP assessment indicated immobilization, but the EMS assessment did not. The kappa for the individual components of the assessments ranged from 0.35 to 0.81, with the kappa for the decision to immobilize being 0.48. The EMS providers' assessments were generally more conservative than the EPs'. CONCLUSION: EMS and EP assessments to rule out cervical spinal injury have moderate to substantial agreement. However, the authors recommend that systems allowing EMS providers to decide whether to immobilize patients should follow those patients closely to ensure appropriate care and to provide immediate feedback to the EMS providers. PMID- 9737406 TI - Evaluating the evaluators: interrater reliability on EMT licensing examinations. AB - INTRODUCTION: Current methods of evaluating the technical competence of Michigan emergency medical technician (EMT) licensure candidates are subjective and potentially unreliable. Evaluators are required to attend a workshop before evaluating practical examination candidates. Despite the workshop, there is too much score variation and not enough observational consistency on the standardized examination. OBJECTIVE: To determine the level of rater reliability for evaluators of EMT practical examinations. METHODS: Data were collected from 104 licensed instructor-coordinators (ICs). Participants watched and scored two practical examinations simulated on videotape, one passing and one failing performance. Variation in student score and level of evaluator agreement concerning skill performance were determined. Michigan's basic EMT practical examination scoring instrument was used. RESULTS: Nine basic EMT, 9 EMT specialist, and 86 EMT-paramedic ICs participated. Thirty-four percent had high school diplomas, 43% associate's degrees, 19% bachelor's degrees, 3% master's degrees, and 1% a doctoral degree. The ICs averaged 14 years of provider experience, and 6.35 years as an IC. The average score for scenario 1 (passing) was 86.4% (SD = 9.15, range = 53). An 80% score is required to pass. Nine ICs (9%) failed the student. Scenario 1 evaluator agreement was 79.4%. The average score for scenario 2 (failing) was 60.9% (SD = 11.57, range = 57). Seventeen ICs (18%) gave the student a passing score. Scenario 2 evaluator agreement was 67.8%. There was no significant difference in student scores based on evaluator level of education, licensure, or evaluator workshop attendance for either scenario. CONCLUSION: Notable variation in scores given by evaluators for a single observed student, combined with low levels of evaluator agreement about skill performance, suggests that evaluators do not reliably rate student performance using the Michigan practical examination instrument. PMID- 9737407 TI - Resident perspectives of EMS as a subspecialty. AB - OBJECTIVE: Emergency medical services (EMS) is frequently considered to be a subspecialty of emergency medicine (EM) despite the unavailability of subspecialty certification. An assessment of future interest in EMS subspecialization and the perceived educational needs of potential EMS physicians was performed in order to provide data to leaders responsible for development of this subspecialty area. METHODS: A survey concerning EMS subspecialization issues was distributed to 2,464 members of the Emergency Medicine Residents Association (EMRA). Questions addressed demographic information, interest in EMS, educational issues, and desired credentials. The response rate was 30% (n = 737). All surveys were analyzed by the Pearson chi-square probability and Mantel-Haenszel tests for linear association. RESULTS: A moderate to very high interest in EMS medical direction was expressed by 84% of the respondents, with 14% interested in full time EMS positions. This interest increased with years of training (p < 0.0001). Almost 89% believed that EMS physicians should have special preparations prior to practice beyond EM residency training. Fewer than half (44%) thought that an EM residency provided sufficient preparation for a significant role in EMS, and this perception increased in intensity with years of training (p < 0.0052). Interest in EMS fellowships (24%) would increase to 36% if subspecialty certification were available (p < 0.0001). Thirty-nine percent believed subcertification should be required of all EMS medical directors if available. CONCLUSIONS: Many EM residents have an interest in active participation in EMS on either a part-time or a full-time basis. Most respondents think EMS is a unique area requiring focused education beyond an EM residency. Interest in EMS fellowships would greatly increase if subspecialty certification were available. PMID- 9737408 TI - Comparison of bag-valve-mask, manually triggered ventilator, and automated ventilator devices used while ventilating a nonintubated mannikin model. AB - OBJECTIVE: To determine whether there were differences in tidal volume (Vt), minute volume (MV), average mask leak per breath (ML), gastric insufflation (GI), and peak airway pressure (PAP) when ventilating a nonintubated mannikin with a bag-valve-mask (BV), manually triggered ventilator (MTV), and automated ventilator (AV). The authors' hypothesis was that there would be no differences among the devices for any of these variables. METHODS: This was a prospective in vitro experimental model. A convenience sample of 19 emergency medical technicians (EMTs) ventilated a nonintubated mannikin-mechanical test lung model with the BV, MTV (flow rate 40 L/min; pressure relief 55 cm H2O), and AV (800 mL/breath; rate 12). Each subject, blinded to volume and pressure gauges, used each device for 2 minutes at both normal (0.1 cm H2O) and poor (0.04 cm H2O) compliances. Vt, MV, GI, and PAP were measured directly and ML was calculated. A survey was issued to the EMTs who participated in the study. Data were analyzed with repeated-measures ANOVA and the Bonferroni-Dunn multiple comparison test with alpha set at 0.05. RESULTS: At the normal compliance, PAP was higher for the BV than the MTV (p = 0.0001) and AV (p < 0.0001). MV was also greater with the BV than with the AV (p = 0.001). PAP was also higher at the poor compliance with the BV than with the MTV and AV (p = 0.008 and 0.013, respectively). The BV had a higher GI at this compliance (p < 0.0001) and a higher ML than the AV (p = 0.002). CONCLUSION: All three devices delivered similar volumes when used by EMTs, but the BV was associated with higher PAP, ML, and GI. PMID- 9737409 TI - The education of out-of-hospital emergency medical personnel in pediatrics: report of a national Task Force. PMID- 9737410 TI - Risk reduction for exposure to blood-borne pathogens in EMS. National Association of Emergency Medical Services Physicians. PMID- 9737411 TI - EMS systems and managed care integration. AB - Emergency medical services systems and MCOs must cooperate and educate each other in order to effect delivery of reliable, high-quality emergency health care to the entire community. Shared goals are rapid access, medically appropriate care, and operational efficiency. An integrated approach is necessary in order to maintain the integrity of EMS systems. EMS systems serve as a safety net for patients with perceived emergencies. Changes in form and function should be guided by outcome studies that ensure the continued delivery of quality emergency health care services. PMID- 9737412 TI - A review of infection control practices, risk reduction, and legislative regulations for blood-borne disease: applications for emergency medical services. PMID- 9737413 TI - Potential liabilities of medical directors for actions of EMTs. PMID- 9737414 TI - An emergency department-based field response team: case report and recommendations for a "go team". PMID- 9737415 TI - The pneumatic anti-shock garment (PASG): can we really recommend it? PMID- 9737416 TI - A critical evaluation of the potential benefits of public access defibrillation. PMID- 9737417 TI - Overview of the preparation, use and biological studies on polyglycerol polyricinoleate (PGPR). AB - The esterification of condensed castor oil fatty acids with polyglycerol gives a powerful water-in-oil emulsifier which is used by the food industry in tin greasing emulsions and as an emulsifier with lecithin in chocolate couverture and block chocolate. A safety evaluation programme was undertaken in the late 1950s and early 1960s to determine whether this food emulsifier polyglycerol polyricinoleate (PGPR). (Quest International trade name ADMUL WOL) presented any health implications for consumers. This programme included acute toxicity tests, subacute rat and chicken toxicity studies, a rat chronic toxicity/multigeneration reproduction study, rodent metabolism, carcinogenicity testing in rat and mouse and a human clinical evaluation. PGPR was found to be 98% digested by rats and utilized as a source of energy superior to starch and nearly equivalent to groundnut oil. There was no interference with normal fat metabolism in rats or in the utilization of fat-soluble vitamins. Despite the intimate relationship with fat metabolism, no evidence was found of any adverse effects on such vital processes as growth, reproduction and maintenance of tissue homeostasis. PGPR was not carcinogenic in either 2-year rat or 80-week mouse feeding studies. The human studies showed no adverse effects on tolerance, liver and kidney function, and fat balance at levels up to 10 g/day PGPR. The acceptable daily intake for PGPR which was set by JECFA in 1974 and the EC/SCF in 1979 is 7.5 mg/kg body weight/day. The UK FAC in 1992 estimated that the maximum per capita mean daily intake of PGPR is 2.64 mg/kg body weight/day. It can be concluded that the use of ADMUL WOL brand of PGPR in tin-greasing emulsions or in chocolate couverture does not constitute a human health hazard. PMID- 9737418 TI - The fate of ingested glyceran esters of condensed castor oil fatty acids [polyglycerol polyricinoleate (PGPR)] in the rat. AB - Samples of the emulsifier polyglycerol polyricinoleate (PGPR) were synthesized using the radiolabelled precursors [1-14C]glycerol ([14C]polyglycerol PGPR), [9,10-3H] or [12-3H]ricinoleic acid ([3H] PGPR) or [1-14C]stearic acid ([14C]stearyl PGPR). The absorption, tissue distribution, metabolism and excretion of these 14C- or tritium-labelled PGPR samples administered to rats was studied. The effects of intestinal and porcine pancreatic lipases on PGPR preparations were examined. Rats were dosed with [1-14C]glycerol, [14C]polyglycerol and ([14C]polyglycerol)PGPR by gavage and their urine. faeces and expired CO2 monitored for 14C. The results from the [1-14C]glycerol treated animals showed extensive metabolism of glycerol. For [14C]polyglycerols, the lower polyglycerols were preferentially absorbed from the intestine and were excreted unchanged in the urine while the higher polyglycerols were found in the faeces. After 4 days, 93% of the dose of polyglycerols was recovered, of which some 30% was found in the urine and 60% in the faeces. Traces of 14C activity were found in depot fat and liver. The excretory pattern and urinary metabolites from ([14C]polyglycerol) PGPR was very similar to that of [14C]polyglycerol. Analysis of urinary and faecal 14C material indicated that the PGPR polymer was digested to give free polyglycerol and polyricinoleic acid. PGPR was synthesised incorporating [1-14C]stearic into polyricinoleic acid which was then esterified with polyglycerol. The resulting [14C]PGPR or [1-14C] stearic acid in a dietary slurry was administered to groups of fed or starved rats by gavage. The results indicated complete digestion of PGPR and absorption of the fatty acids. The 14C material absorbed was extensively laid down in depot fat and some metabolism to 14CO2 was demonstrated. The fate of the stearic acid was similar whether dosed alone or incorporated into the PGPR polymer. Samples of PGPR were synthesized containing 3H-labelled ricinoleic acid. The resulting [3H]PGPR was intubated into rats as a component of a dietary slurry. The results indicated that the polymer is extensively digested and 90% of the administered tritium is absorbed. The absorbed material was extensively metabolized within 24 hr so that large amounts of tritium were present in the aqueous phase of the tissues examined. After 24 hr, less than 5% of the administered material was present as lipid material, of which a large proportion was as non-hydroxy fatty acids. No traces of polymer material were found in the tissues examined. In vitro digestion of PGPR by porcine pancreatic lipase and rat intestinal fractions was demonstrated. The results indicate very extensive digestion of the PGPR polymer to polyglycerols and fatty acids. The fatty acids are metabolized extensively. The mono-, di- and triglycerols are extensively absorbed from the intestinal tract and rapidly excreted in the urine unchanged but the hexa-, penta- and higher polyglycerols are essentially not absorbed and excreted in the faeces unchanged. PMID- 9737419 TI - A three-generation reproduction study on polyglycerol polyricinoleate (PGPR) in Wistar rats. AB - A series of toxicology studies were conducted in the 1950s and 1960s to investigate the toxicity of ADMUL WOL, a brand of polyglycerol polyricinoleate (PGPR). Included as part of these investigations was a three-generation reproduction study in rats. The control rats received a commercial pelletted stock diet and the treated rats were given the same diet ground with 1.5% (w/w) PGPR. A continuous breeding protocol was adopted, in which the breeding pairs were maintained until the female had produced five litters or when it became evident that breeding had ceased. The main focus of the study design was to observe any effect on breeding. The parameters measured in each of the three generations included number of litters per dam, average litter size, average weaning weights of males and females, litters per group showing 100% survival and total survival (%) at day 21. Growth was normal throughout the three generations and there were no deaths or clinical signs associated with the consumption of PGPR. The only significant change in breeding performance was a reduction in the percentage of animals weaned in the second generation, but as this occurred in the control group to a similar extent it was concluded that this was due to an unknown environmental factor and was not treatment related. A histological examination of selected tissues from those rats continued for 1 year failed to show any lesions which could be ascribed to the consumption of PGPR. In conclusion, rats fed 1.5% (w/w) PGPR showed no evidence of a cumulative effect on breeding performance over three generations. PMID- 9737420 TI - Human studies on polyglycerol polyricinoleate (PGPR). AB - A series of toxicology studies was conducted in the 1950s and 1960s to investigate the toxicity of ADMUL WOL, a brand of polyglycerol polyricinoleate (PGPR). A component of these investigations included studies in human subjects. During 1964 and 1965, PGPR was fed to 19 human volunteers whose diet contained constant levels of fat and protein. Up to 10 g/day PGPR was fed to each volunteer in soups, cakes and toffee bars for 2 weeks. Pre-exposure normal values of biochemical parameters were established. Fat balance tests confirmed that digestion and absorption of PGPR took place. No consistent effect of PGPR on the various biochemical parameters was observed, nor had PGPR any toxic effect on liver and kidneys. The consumption of PGPR by humans produced no adverse effects. The quantities consumed, up to 10g/day, was equivalent to approximately 63 times the estimated maximum per capita mean daily intake by man of 2.64 mg kg body weight/day. It is therefore concluded from this study that the consumption of ADMUL WOL, a brand of PGPR, has no adverse effects in man. PMID- 9737421 TI - Assessment of the carcinogenic potential of polyglycerol polyricinoleate (PGPR) in rats and mice. AB - The carcinogenic potential of the food emulsifier ADMUL WOL brand of polyglycerol polyricinoleate (PGPR) was evaluated in rats and mice. Groups of 60 male and 60 female rats were given purified diets containing 5% of either PGPR or groundnut oil for 2 years. Groups of 25 male and 25 female mice were given purified diets containing 5% of either PGPR or groundnut oil for 80 weeks. No carcinogenic effect of PGPR was observed. In addition, dietary PGPR had no adverse effect on growth, food consumption, longevity and haematology. Organ weight analysis revealed an increase in liver and kidney weight in both male and female rats and female mice. Histological analysis of tissues revealed no treatment related adverse effects. PMID- 9737423 TI - Effects of the garlic components diallyl sulfide and diallyl disulfide on arylamine N-acetyltransferase activity in human colon tumour cells. AB - Diallyl sulfide (DAS) and diallyl disulfide (DADS), major components of garlic, were used to determine inhibition of arylamine N-acetyltransferase (NAT) activity in a human colon tumour (adenocarcinoma) cell line. Two assay systems were performed, one with cellular cytosols (9000g supernatant), the other with intact bacterial cell suspensions. The NAT activity in a human colon tumour cell line was inhibited by DAS and DADS in a dose-dependent manner in both system: that is, the greater the concentration of DAS and DADS in the reaction, the greater the inhibition of NAT activities in both systems. The data also indicated that DAS and DADS decrease the apparent values of Km and Vmax of NAT enzymes from human colon tumour cells in both systems examined. This is the first report to demonstrate that garlic components do affect human colon tumour cell NAT activity. PMID- 9737422 TI - Reaction of cyclohexylamine with hypochlorite and enhancement of oxidation of plasma sulfhydryl groups by hypochlorite in vitro. AB - In this study we investigated the reaction of cyclamate and its major metabolite, cyclohexylamine (CyhNH2), with NaOCl. NaOCl at 100 microM was allowed to react with various concentrations of cyclamate and CyhNH2, and the reactivity was compared with those of reduced glutathione (GSH) and ascorbic acid. The results showed that CyhNH2 was less reactive with NaOCl than GSH but was slightly more reactive than ascorbic acid at concentrations below 50 microM. CyhNH2 at 75 and 100 microM did not further decrease NaOCl. Cyclamate was much less reactive than CyhNH2, with only 43% loss in NaOCl at 100 microM cyclamate. When human blood plasma was incubated with 0.75 microM NaOCl, inclusion of CyhNH2 enhanced oxidation of sulfhydryl groups in a concentration-dependent manner, with complete oxidation of SH groups at 7.5 mM CyhNH2. Cyclamate had no effect. This enhancement by CyhNH2 suggests the formation of reactive products from the reaction of CyhNH2 with NaOCl. Absorption spectra demonstrated that reaction of CyhNH2 with NaOCl at pH 7.4 produced N-monochloramine, as evidenced by the appearance of a new peak at 245 nm and by the disappearance of the 292-nm peak of NaOCl. Cyclamate, which contains a sulfamic acid instead of a primary amine, also reacted with NaOCl at pH 7.4, but the reaction was much less pronounced and the product was probably not monochloramine since the peak was at 270 nm rather than at 245 nm. Because cyclamate is an important sweetener in many countries for people with diabetes mellitus, the possibility exists that CyhNH2 may enhance oxidation of important proteins by HOCl/OCl-. PMID- 9737425 TI - Safety evaluation of amino peptidase enzyme preparation derived from Aspergillus niger. AB - An amino peptidase enzyme preparation obtained from Aspergillus niger was subjected to a series of toxicological tests to document the safety for use as a processing aid for food. The enzyme preparation was examined for subacute and subchronic oral toxicity, and mutagenic potential. No evidence of oral toxicity or mutagenicity was found. Administration of the amino peptidase enzyme preparation at doses of 500, 1000 and 2000 mg/kg body weight/day for 90 days did not induce noticeable signs of toxicity. The no-observed-adverse-effect level (NOAEL) of the enzyme preparation in the subchronic toxicity study was 2000mg/kg body weight/day (equivalent to 1152 PHEA units/kg body weight/day). It can be concluded that no safety concerns were identified in the studies conducted with this amino peptidase enzyme preparation derived from Aspergillus niger and produced under controlled fermentation conditions. PMID- 9737424 TI - The ability of the rat to metabolize myristoyl-methionine: an acylamino acid with potentially useful antibacterial properties. AB - Two experiments with Sprague Dawley rats tested their ability to hydrolyse myristoyl-methionine (M-M) into myristic acid and L-methionine (M). In the first experiment, lasting for 3 days. male rats were orally administered [9,10 3H]myristoyl-L-[35S]methionine. The recovery of radioactivity was approximately 90% for both isotopes; 19% of the administered 3H was recovered in the urine and 16% in the faeces, while the recovered 35S activity was 13 and 12%, respectively. The balance of the radioactivity was found among the tissues, organs and blood. In the second experiment, male and female rats received soybean-based diets which were supplemented with either 0.305% M-M or 0.2% M (both diets contained equal amounts of M) for periods up to 4 weeks. The growth rate of the rats receiving the 0.305% M-M diets was slightly slower than that for the rats on the 0.2% M diet, but the difference was not statistically significant (P > 0.05). The M-M rats had a transitory decrease in feed consumption, suggesting that palatability may have contributed to the growth difference and that a somewhat greater amount of M-M was necessary for the rat to attain the same growth rate as that produced by 0.2% M. When the amount of dietary M-M was increased to 3.05% M-M, a greater reduction in feed consumption and body weight gain was observed. This latter diet was an initial attempt to study the potential toxicity of M-M. None of the haematological, clinical chemistry or organ weight data suggested that M-M was overtly toxic per se, but longer-term feeding studies are needed to evaluate the potential toxicity of M-M more fully. PMID- 9737426 TI - Characterization of cell-cycle arrest by fumonisin B1 in CV-1 cells. AB - Fusarium moniliforme is a widespread fungal pathogen which primarily infects corn, but can also infect rice or wheat. Fusarium moniliforme produce several mycotoxins, the most prominent of which is called fumonisin B1 (FB1). Epidemiological studies have indicated that ingestion of fumonisins correlates with a higher incidence of oesophageal cancer in Africa and China. Fumonisins also cause a neurodegenerative disease in horses, induce hepatic cancer in rats, are nephrotoxic in rats, or cause pulmonary oedema in swine. Structurally, fumonisins resemble sphingolipids and can alter sphingolipid biosynthesis. suggesting that sphingolipid alterations play a role in disease and carcinogenesis. Previous studies determined that FB1 blocked cell-cycle progression in CV-1 cells but not COS-7 cells. Herein, we have examined the effects that FB1 treatment has on cell-cycle regulatory proteins. Our studies established that FB1 treatment of CV-1 cells, but not COS-7 cells, leads to dephosphorylation of the retinoblastoma (Rb) protein. Cyclin dependent kinase 2 (CDK2) activity was repressed five- to 10-fold and cyclin E protein levels were lower in CV-1 cells after fumonisin treatment. Two CDK inhibitors, Kip1 and Kip2, were induced within 3 hours after fumonisin treatment of CV-1 cells, suggesting these two proteins mediate cell-cycle arrest induced by FB1. This mycotoxin caused large increases in sphinganine within 3 hours after addition of FB1. As sphingoid bases are known to induce Rb phosphorylation, this increase in sphinganinie might be the stimulus for the suppression of cyclin dependent kinase activities via Kip1 and Kip2. The ability of FB1 to accumulate sphingosine or sphinganine and arrest the cell cycle in some cells but not others may play an important role in carcinogenesis or disease. PMID- 9737427 TI - Desmutagenicity of milk cultured with Lactobacillus acidophilus strains against mutagenic heated tauco. AB - Desmutagenicity of milk cultured with Lactobacillus acidophilus strains on the mutagenicity of heated salty and sweet tauco were examined using streptomycin dependent (SD) 510 strain of Salmonella typhimurium TA98 as tester culture. Cultured milk samples widely exhibited desmutagenic effects against mutagenic heated salty and sweet tauco. Mutagenicity of heated salty tauco was inhibited by acidophilus cultured milks stronger than that of heated sweet tauco. Milk cultured with strains SBT2054, SBT0299, SBT0274, SBT10238. SBT1702, SBT10240, SBT10241 and SBT10239 showed high inhibition against the mutagenicity of both heated salty and sweet taucos. Maximum inhibition was reached after 24 hr of incubation which corresponded to stationary growth phase. Desmutagenic activities of the acidophilus cultured milks against mutagenic heated tauco were mainly attributed to the bacterial cells and also to casein of milk. PMID- 9737428 TI - Mutagenicity and DNA-damaging activity of decomposed products of food colours under UV irradiation. AB - Five synthetic food colours Food Red Nos 3, 40 and 102 and Food Blue Nos 1 and 2, and their UV irradiated products were tested for mutagenic activity by means of the Ames test using Salmonella typhimurium strains TA98 and TA100. Food colours were irradiated with UV light for 14 days. Food Red Nos 3, 40 and 102 and Food Blue No. 1 were non-mutagenic before and after irradiation. UV irradiated products of Food Blue No. 2 were mutagenic in TA98 with or without S-9 mix. The mutagenic activity increased with increasing irradiation period, reached maximum potency on day 6, and then decreased. Moreover, Food Blue No. 2 showed DNA damaging activity after 14 days of irradiation in rec-assay using Bacillus subtilis strains H17 and M45. The capillary electrophoresis was applied for the analysis of UV irradiated products of Food Blue No. 2. The original peak of Food Blue No. 2 was decomposed into seven peaks after UV irradiation. PMID- 9737429 TI - Decrease of manganese superoxide dismutase activity in rats fed high levels of iron during colon carcinogenesis. AB - Diets high in fat or iron have been associated with an increased risk for development of colon cancer. These two dietary factors are known to decrease manganese superoxide dismutase (MnSOD) activity in colonic mucosa. MnSOD is an antioxidant enzyme that protects mitochondria from oxygen radical damage. MnSOD has tumour suppressive activity and is absent or decreased in most tumours, including those from the colon. This study was designed to determine the effects of high dietary lipid and iron levels on MnSOD activity during the early weeks of colon carcinogenesis. Male Fischer-344 rats were fed 20% lipid diets of either corn oil or menhaden oil containing adequate iron (35 mg/kg) or supplemental iron (535 mg/kg). Rats from each diet were divided into carcinogen treatment groups and given two weekly injections of either azoxymethane (AOM) at a dose of 12 mg/kg, or saline. Mucosal tissue was collected 1, 6 and 12 wk following injections and analysed for MnSOD activity, mineral concentration and nuclear aberrations. Results showed that iron supplementation increased nuclear aberrations, and decreased manganese concentration and MnSOD activity in colonic mucosa ot control animals. AOM, and interaction of iron and AOM, also decreased MnSOD activity. A decrease in the activity of this enzyme during carcinogenesis may be one mechanism whereby these dietary factors ultimately increase tumour risk. PMID- 9737430 TI - Acute toxicity of ethylene glycol mono-n-butyl ether in the guinea pig. AB - Acute toxicity values, such as oral and percutaneous LD50s, are often used as the basis for classifying chemicals into toxicity categories, and their subsequent regulation. Such values obtained for ethylene glycol mono-n-butyl ether (EGBE; 2 butoxyethanol) in rats and rabbits indicate that it is moderately toxic. However, the cause of death in these acute studies appeared to be secondary to acute intravascular haemolysis, an effect for which guinea pigs and humans are much less sensitive than rats, mice and rabbits. Recently-conducted acute toxicity studies in the guinea pig resulted in an acute oral LD50 of 1400 mg/kg, an acute percutaneous LD50 of greater than 2000 mg/kg, and a 1-hr LC50 greater than 633 ppm. These data are compared with published acute toxicity values, and indicate that the predicted acute toxicity of EGBE in humans, based on data from the guinea pig, would be less than that observed in other animal species. Based in part on the guinea pig data, EBGE is no longer classified as a poisonous substance by either the United Nations or US Department of Transportation. PMID- 9737431 TI - The relationship among microsomal enzyme induction, liver weight and histological change in rat toxicology studies. AB - The purpose of this study was to determine what histological changes, if any, accompany liver enlargement and microsomal enzyme induction in rats administered high doses of therapeutic agents in preclinical toxicology studies. This was accomplished by evaluating a database derived from a series of 11 induction studies in rats with 10 novel compounds comprising five therapeutic classes. Results from serum enzyme chemistry analyses, gross organ weight changes, and histological analyses of the liver sections were evaluated and compared with the magnitude and extent of hepatic cytochrome P450 induction. All compounds were administrated via oral intubation once a day for the duration of the study using multiple doses, each proportionally based on body weight. During the course of these studies, serum clinical chemistry data and clinical observations were recorded. After necropsy, histopathology observations were made, and hepatic microsomes were assayed for cytochrome P450 content and associated drug metabolizing enzymes. In some cases, cyanide-insensitive beta-oxidation of palmitoyl CoA was also assayed. Liver weight increases of 20% or greater were associated with histological evidence of hypertrophy, but neither the severity of hypertrophy nor the magnitude of liver weight increase correlated with the magnitude of drug-metabolizing enzyme elevations. Hypertrophy alone was not associated with serum enzyme increases. While there was a correlation between the incidence of increased liver weights and microsomal enzyme induction, the magnitudes of these increases were not related. Decreased serum triglycerides were often associated with elevated beta-oxidation attributed to hepatic peroxisome proliferation. It was concluded that, while slight ALT elevations occasionally were observed, hepatic microsomal enzyme induction was generally not accompanied by substantial morphological changes or elevated serum enzyme levels considered indicative of liver injury. PMID- 9737432 TI - Formation of malonaldehyde in the presence of probucol, an anti-atherosclerosis drug. AB - Formation and inhibition of malonaldehyde (MA) from blood plasma lipids oxidized by Fenton's reagent in the absence or presence of probucol [4,4' (isopropylidenedithio)bis(2,6-di-tert-butylphenol)] and L-ascorbic acid were investigated. The amount of MA formed was quantitatively analysed by gas chromatography. L-Ascorbic acid inhibited MA formation by about 30% at the level of 4.0/micromol, but the amount of MA formed was increased by the presence of probucol. When 3.0 micromol oxidized probucol was hydrolysed at pH 1. 3 and 5, 2616.5 nmol, 287.5 nmol and 103.9 nmol MA were recovered, respectively. This is the first report of quantitative analysis of MA formed from probucol on oxidation. PMID- 9737433 TI - Risk assessment of dithiocarbamate accelerator residues in latex-based medical devices: genotoxicity considerations. AB - The Medical Devices Agency (MDA) has investigated potential human health hazards arising from the presence of dithiocarbamate vulcanization accelerators in latex products (mainly gloves). After collection of manufacturer's data on usage and residues of these accelerators, an independent investigation of solvent extractable residues and dithiocarbamate migration into aqueous simulants was commissioned, to complement equivalent "in-house" test data from two major manufacturers. The presence of extractable accelerator residues in commercial products was confirmed. Potential human health hazards associated with dithiocarbamates include genotoxicity and possible carcinogenicity: a review of published data was conducted to evaluate the evidence for this, with particular reference to three zinc dithiocarbamates with significant commercial usage (ZDMC, ZDEC and ZDBC: see Fig. 1). Data gaps were identified, and mutagenicity studies commissioned to fill these. These studies comprised tests both in vitro (bacterial and L5178Y cell gene mutation, cultured lymphocyte chromosome aberration) and in vivo (mouse bone marrow micronucleus, rat liver UDS). It is concluded that ZDMC must be considered a genotoxin (and thus a probable carcinogen): residues of this substance in latex medical devices should be minimized. ZDEC proved genotoxic in vitro but was not clearly genotoxic in vivo, and may have activity intermediate between that of ZDMC and that of ZDBC, which showed at most weak activity in a single in vitro (chromosome aberration) test. It is proposed that the use of ZDBC as a vulcanization accelerator in the manufacture of latex gloves, rather than ZDEC, ZDMC or their precursors, would reduce or remove the health concerns arising from accelerator residues. PMID- 9737434 TI - A hairless mouse model for assessing the chronic toxicity of topically applied chemicals. AB - An enormous number of synthetic chemicals are incorporated in topical drugs, cosmetics and toiletries. These have the potential to cause irritant reactions when chronically applied to human skin. In predictive tests for assessing the irritancy potential of these chemicals, haired species, especially rabbits, guinea pigs and mice, have figured prominently. Customarily these tests, including the renowned Draize rabbit test, have entailed a single acute exposure or at most daily exposures over a few weeks. Estimation of inflammation and tissue injury in these models have relied on visual assessment. We submit that this approach is no longer acceptable. Visual assessments are unreliable. Reactions which are scored equivalently by the naked eye may differ strikingly when examined histologically. Moreover, tissue injury may be present in clinically normal skin. Short-term results. even when abetted by routine histological evaluations, cannot predict the degree of injury from long-term exposures. Cosmetics and toiletries, for example, are used daily for decades, often over most of the lifespan of persons who are well groomed. We present the hairless mouse as a convenient, reliable model for assessing the chronic toxicity of diverse chemicals. Histological examination enables a detailed description of the different tissue components which participate in the complex cascade of changes that comprise the inflammatory response. PMID- 9737435 TI - Carcinogenic heterocyclic amines in model systems and cooked foods: a review on formation, occurrence and intake. AB - Frying or grilling of meat and fish products may generate low ppb levels of mutagenic/carcinogenic heterocyclic amines (HAs). Many heterocyclic amines are formed via the Maillard reaction from creatine, free amino acids and monosaccharides; compounds naturally occurring in protein-rich foods of animal origin. The formation and yield of HAs are dependent on physical parameters, such as cooking temperature and time, cooking technique and equipment, heat and mass transport, and on chemical parameters, especially the precursors to HAs. This paper reviews the current knowledge on the formation of HAs in cooked foods and model systems, and summarizes data on the content of HAs in various cooked foods, and estimates of the dietary intake of HAs. It should be noted that the presence of carcinogens of other types in food (e.g. nitrosamines, aromatic amines, cholesterol oxide products) and that their generation during frying and grilling are outside the scope of this review. PMID- 9737436 TI - Role of new and established antiepileptic drugs. PMID- 9737437 TI - Awareness and responsiveness during partial seizures. AB - We examined the impairment of consciousness during partial seizures (PS) arising from various brain sites according to the operational definition of the international classification, i.e., altered awareness and/or responsiveness. The subjects were 142 patients who underwent intracranial EEG evaluation and subsequent resective surgery. First, the patients were examined to determine whether they usually had been partially or completely aware of their seizures. Second, spontaneous habitual seizures that had been videotaped with simultaneous intracranial EEG recording were reviewed to determine responsiveness and recall during ictal behavioral alterations. In all, 114 patients were partially or completely aware of their seizures. Patients who tended not to be aware of their seizures were those with frontal lobe epilepsy (FLE) with extensive epileptogenic regions on the language nondominant side and those with temporal lobe epilepsy (TLE) with seizure origin in the lateral cortex of the language dominant side. Of the 21 patients with FLE, 88 with TLE, and 4 with occipital lobe epilepsy, 7, 22, and 2 patients responded to stimuli during the seizure, respectively, but only 11 of the patients with FLE and none of the other patients could recall the stimuli applied during the behavioral alterations. Bilateralization of seizure discharges correlated with impaired responsiveness. According to the International Classification, about half of patients with FLE had only simple partial seizures (SPS) and the other patients had complex partial seizures (CPS). Altered awareness and/or responsiveness occurred in most habitual partial seizures in our subjects. The term "complex" appears to be useful in clinical practice, although the contents of ictal behavior and the site or side of seizure origin are not implied. PMID- 9737438 TI - Epilepsy and recursive consciousness with special attention to Jackson's theory of consciousness. AB - John Hughlings Jackson's theory of consciousness has been reconsidered. The author stressed that his uniqueness as a neuroscientist lay in his keen interest in the recursive nature of human consciousness. Two clinical symptoms of interest to Jackson were discussed: recurrent utterances and mental diplopia. Recurrent utterances were believed to represent an exceptional state, in which the unconscious process in speech production, otherwise destined to be swept away automatically, became manifest and observable. Jackson regarded mental diplopia as a revelation of otherwise inaccessible duality of all healthy mental actions. Therefore, he supposed that the essence of recursive consciousness resided in a transformation of multiple, multidirectional, unconscious processes into a linear, unidirectional process. PMID- 9737439 TI - Impairment of consciousness during epileptic seizures with special reference to neuronal mechanisms. AB - Two neuronal structures, i.e., the cerebral cortex and the subcortical structures, were shown by clinical observations to be involved in maintaining consciousness. The alteration of consciousness during epileptic seizures is discussed with respect to these findings: Alterations of consciousness during epileptic seizures may be produced by subcortical, i.e., reticular formation, and/or cortical dysfunction followed by excessive, hypersynchronous neuronal discharges. An impairment of consciousness during absence seizures may be due mainly to cortical dysfunction; during complex partial seizures (CPS), it may be due to dysfunctional subcortical neuronal structures. The mechanisms underlying an alteration of consciousness are defined as causing "irritative" functional disturbances and/or as having "inhibitory" effects on consciousness-related structures. PMID- 9737440 TI - Epileptic seizures and pseudoseizures from the viewpoint of the hierarchy of consciousness. AB - Epileptic seizures and pseudoseizures in temporal lobe epilepsies were studied from the viewpoint of the hierarchy of consciousness. Twenty-two patients with temporal lobe epilepsies (TLE) who showed true amnesia or impairment of consciousness developing from the dreamy state, even though their actions and movement continued, were selected among 160 patients with TLE, nine patients with manifested pseudoseizures, pseudoseizure status, and complex partial seizure status (CPSE) were investigated. Twelve patients in whom impairment of consciousness followed the dreamy state recognized their own existence and maintained some self-directed consciousness. The other 10 patients with amnesia were aware of their goals. Furthermore, pseudostatus ranged from epileptic seizures during pseudoseizure status to pseudoseizures during status epilepticus (SE). In some cases of CPS, awareness and self-directed consciousness were only partially pseudoseizures, disorders of self-directed consciousness are assumed to influence awareness and arousal. PMID- 9737442 TI - West syndrome and its related epileptic syndromes. AB - The purpose of this project was to study the relationship between West syndrome (WS) and its related epileptic syndromes, and reconsider the nosological limits of WS. The electroclinical features of 45 patients who experienced spasms in series were investigated, as well as some features not common in patients with WS. All patients were mentally retarded. The patients were divided into three groups: Group 1 consisted of 12 patients with refractory epilepsies with onset in early infancy, group 2 consisted of 5 patients with symptomatic localization related epilepsies associated with spasms in series, and group 3 consisted of 28 patients with generalized epilepsies who had spasms in series after age 2 years. Partial seizures were the dominant symptom throughout the clinical course and spasms in series associated with atypical hypsarrhythmia appeared transiently during infancy in a significant number of the patients in group 1. In group 2, complex partial seizures (CPS) were the main seizure type and hypsarrhythmia was not observed during the clinical course. The EEGs in group 3 patients showed diffuse slow spike-waves or multifocal epileptic discharges in all but 1 patient. The EEG of the remaining patient still showed hypsarrhythmia at age 8 years. Therefore, group 1 patients should be classified as having WS although cortical mechanisms play a critical role in the occurrence of their seizures. Group 2 patients should be considered as having a type of localization-related epilepsy even though they share a similar pathophysiological mechanism with group 1. In group 3, 1 patient whose EEG still showed hypsarrhythmia was classified as having WS. The other patients should be classified as having generalized epilepsies other than WS. PMID- 9737441 TI - Heterogeneity of ictal SPECT findings in nine cases of West syndrome. AB - We evaluated the ictal and interictal single photon emission computed tomography (SPECT) of 9 patients with West syndrome (WS). In this group, we noted two clear patterns of cortical hyperperfusion and subcortical hyperperfusion in the ictal SPECT. Both patterns were different from the previously documented ictal patterns for complex partial seizures (CPS) or secondarily generalized seizures. Our results suggest that the tonic spasms of WS do not always have a single neurophysiological basis; e.g., patients with hemihypsarrhythmia and focal hypsarrhythmia did not show ictal hyperperfusion of the lesion with hypsarrhythmia. These findings indicate that the origin of hypsarrhythmia as an EEG feature and the origin of tonic spasms may be different in such patients. In particular, hypsarrhythmia appears to originate from cortical lesions, whereas the subcortical structures may be primarily responsible for the tonic spasms. Our report is the first published study of ictal SPECT in patients with WS. PMID- 9737443 TI - Antiepileptic drug treatment of West syndrome. AB - We sent questionnaires about current therapy for West syndrome (WS) to 208 institutions at which the pediatric members of the Japan Epilepsy Society were in residence. One hundred twenty-nine institutions (62%) responded. Thirty-eight institutions (29.5%) treated symptomatic and cryptogenic WS differently. Vitamin B6 was the most preferred first-line drug, followed by the combination of vitamin B6 and valproate (VPA) or monotherapy with VPA. ACTH was the third choice among drugs. The combination therapy of vitamin B6 and VPA was effective in 58.3% of the patients with symptomatic WS. PMID- 9737444 TI - ACTH therapy for infantile spasms: a combination therapy with high-dose pyridoxal phosphate and low-dose ACTH. AB - Combination therapy consisting of high-dose pyridoxal phosphate (40-50 mg/kg/day) and low-dose synthetic ACTH (0.01 mg/kg/day) was prescribed in 28 children with infantile spasms. Monotherapy with pyridoxal phosphate provided excellent seizure control in 3 of the 28 (11%) patients. ACTH was subsequently added to the regimen of the remaining 25 patients. As of 1 month after discontinuing the ACTH treatment, 21 of the 25 (84%) patients had experienced no seizures. The mean interval until seizure control was achieved was 4.1 days after the start of treatment with ACTH. The 21 patients have been monitored for a mean of 34.9 months (range 2-81 months); 6 patients (29%) have had recurrences of infantile spasms, and 10 (48%) have experienced normal development. Fourteen of the 28 patients (50%) have had transient increases in liver enzymes, but none of the patients developed more serious side effects. PMID- 9737445 TI - Evolutional changes and outcome of West syndrome: correlation with magnetic resonance imaging findings. AB - The prognosis and evolutional changes of 77 patients with West syndrome (WS) were studied after patients were classified into four groups on the basis of their magnetic resonance imaging (MRI) findings: anomaly, perinatal injury, normal, and the other groups. The average age at onset of spasms was earliest in the patients with anomalies and latest in patients with normal MRI findings. Patients with normal MRI findings had the shortest duration of spasms, and patients with anomalies had the longest duration of spasms. Antecedent seizures were observed in 6 patients (3 patients with anomalies, 1 patient with normal MRI findings, and 2 patients with other abnormalities). Thirty-five patients had subsequent seizures. Patients with anomalies often had partial seizures and patients with perinatal injuries often had generalized seizures. Seizures were infrequent in patients with normal MRI findings. Developmental outcome was best in the patients with normal MRI findings and worst in patients with perinatal injuries. Various types of epileptic syndromes occurred subsequent to WS in patients with anomalies, although nonspecific symptomatic generalized epilepsy was common in patients with perinatal injuries. These results suggest that seizure prognosis, evolutional changes in seizures, and developmental outcome are different among the types of brain lesions. PMID- 9737446 TI - Long-term course of West syndrome associated with tuberous sclerosis. AB - We retrospectively analyzed the long-term clinical profiles of 47 patients with West syndrome (WS) associated with tuberous sclerosis (TS) the follow-up study for >10 years showed that WS developed into epilepsies of various types in all patients. Their final diagnoses were symptomatic generalized epilepsy (SGE: 29 patients, 62%), symptomatic partial epilepsy (PE: 14 patients, 30%), and undetermined epilepsy (UE: 4 patients, 8%). Eighty-five percent of all patients continued to show the same types of epilepsy that they had at their first examination, but in 15% the diagnosis had to be revised at the end of the follow up period. The outcome of seizures was unfavorable for the SGE group. In the PE group, however, seizures were controlled in 9 (64%) of the 14 patients, and disturbances of daily activity were mild. In patients with WS associated with TS, the seizure outcome and social activity varied depending on the type of epilepsy or epileptic syndrome that developed from WS. The complicated clinical developments from WS with TS multiple cortical tubers related to epileptogenesis in addition to impairment of brain development. PMID- 9737447 TI - Presidential address: trials of the Southern Association for Vascular Surgery. PMID- 9737448 TI - A comparison of carotid angioplasty with stenting versus endarterectomy with regional anesthesia. AB - INTRODUCTION: Percutaneous transluminal angioplasty with stenting (PTAS) has been considered a potential alternative to carotid endarterectomy (CEA) for stroke prevention. Interventionalists have suggested that PTAS carries less anesthetic risk than CEA. The treatment of carotid stenosis with local or regional anesthesia (LRA) allows direct intraprocedural neurologic evaluation and avoids the potential risks of general anesthesia. METHODS: We retrospectively analyzed the clinical charts of 377 patients who underwent 414 procedures for the elective treatment of carotid stenosis in 433 cerebral hemispheres with LRA between August 1994 and May 1997. Group I (312 hemispheres) underwent PTAS, and group II (121 hemispheres) underwent CEA. RESULTS: The indications for treatment included the following: asymptomatic severe stenosis (n = 272; 62.8%), transient ischemic attack (TIA; n = 100; 23.1%), and prior stroke (n = 61; 14.1%). The early neurologic results for the patients in group I (n = 268) included 11 TIAs (4.1%), 23 strokes (8.6%), and 3 deaths (1.1%). The early neurologic results for the patients in group II (n = 109) included 2 TIAs (1.8%), one stroke (0.9%), and no deaths. The total stroke and death rates were 9.7% for the patients in group I and 0.9% for the patients in group II (P = .0015). The cardiopulmonary events that led to additional monitoring were evident after 96 procedures in group I (32.8%) and 21 procedures in group II (17.4%; P = .002). CONCLUSION: PTAS carries a higher neurologic risk and requires more monitoring than CEA in the treatment of patients with carotid artery stenosis with LRA. The proposed benefit for the use of PTAS to avoid general anesthesia cannot be justified when compared with CEA performed with LRA. PMID- 9737449 TI - Gastrointestinal complications after aortic surgery. AB - BACKGROUND AND PURPOSE: A major gastrointestinal complication (GIC) after aortic surgery may be disastrous, but these complications have received scant attention. This study was performed to determine the risk factors, associated events, and outcomes for patients with GIC. METHODS: We performed a secondary analysis of a prospective study that examined 120 consecutive patients who underwent transperitoneal aortic revascularization for aneurysmal or occlusive disease. RESULTS: The following 29 GICs developed in 25 patients (21%) within 30 days of aortic surgery: paralytic ileus that required replacement of nasogastric tubes (n = 12), upper gastrointestinal bleeding (n = 5), Clostridium difficile enterocolitis (n = 5), acute cholecystitis (n = 2), mechanical obstruction (n = 2), ascites (n = 2), and colon ischemia (n = 1). Seven patients required operations for GICs after aortic revascularization. A comparison of patients with and without GICs showed no differences in the prevalence of risk factors, presence of mesenteric artery stenoses, coexisting medical illnesses, antecedent gastrointestinal history, operative indication, preoperative fluid administration, or duration of operation. However, patients with GICs had more intraoperative complications (P = .004), greater intraoperative blood loss (P = .02), and more fluids during the postoperative period (P = .008). The mean duration of mechanical ventilation was 71 +/- 23 hours for patients with GICs versus 7 +/- 2 hours for patients without GICs (P = .006). A higher prevalence of pulmonary (P = .004) and renal (P = .001) complications was seen in the patients with GICs. The mean stay in the intensive care unit was 16 +/- 2 days for patients with GICs as compared with 5 +/- 0.4 days for patients without GICs (P < .001). Four deaths occurred, all caused by multisystem organ failure: 3 patients had GICs, and 1 did not have a GIC (P = .007). CONCLUSIONS: These results show that GICs are prevalent in transperitoneal aortic surgery and are associated with severe morbidity rates, increased hospital costs because of prolonged stay, and increased mortality rates. Some GICs appear to be associated with intraoperative events that lead to visceral hypoperfusion, and others can be attributed to mechanical causes. However, none of the variables examined in this study were predictive of GICs. In all, GICs should be considered serious adverse sequela after aortic revascularization. Because no risk factors for GICs have been identified, these complications currently cannot be prevented. PMID- 9737450 TI - Surgical repair of ruptured abdominal aortic aneurysms in the state of Maryland: factors influencing outcome among 527 recent cases. AB - PURPOSE: Abdominal aortic aneurysm (AAA) rupture has been historically associated with high operative mortality rates. In this community-based, cross-sectional study, we examined factors influencing outcome after operations performed for ruptured AAA (rAAA). METHODS: An analysis of a state database identified 3820 patients who underwent AAA repair between 1990 and 1995, including 527 (13.8%) who had an operation for an rAAA. Demographic variables examined included patient age, gender, race, associated comorbidity rates, operative surgeon experience with rAAA, and annual hospital rAAA and total AAA operative volumes. Outcomes measured included operative mortality rates, hospital length of stay, and charges. RESULTS: Operative mortality rates increased significantly with advancing age (P < 0.0001) but were not related to gender (P = 0.474) or race (p = 0.598) and were significantly lower among patients with hypertension (P = 0.006) or pulmonary disease (P = 0.045). There was no relationship between hospital rAAA or total AAA volume and rAAA repair mortality rate, although high volume surgeons (i.e., performing more than 10 rAAA repairs) had decreased mortality rates and hospital charges compared with other surgeons. Hospital lengths of stay and charges increased with age among survivors, but not nonsurvivors, of rAAA repair. Despite a stable incidence of rAAA repairs during the study interval and no significant change in the mean age of patients undergoing operation or the percentage of operations performed by high-volume surgeons, the statewide mortality rate declined from 59.3% to 43.2% (P = 0.039). CONCLUSION: The incidence of rAAA does not appear to be declining. Although operative rAAA repair continues to be associated with substantial risk and remains an especially lethal condition among the elderly, the operative mortality rate has declined in recent years in Maryland. Lower operative mortality rates and hospital charges are associated with operations performed by high-volume surgeons. PMID- 9737451 TI - Continued expansion of aortic necks after endovascular repair of abdominal aortic aneurysms. EVT Investigators. EndoVascular Technologies, Inc. AB - BACKGROUND: Longitudinal studies have revealed that the aortic segment proximal to an infrarenal abdominal aortic aneurysm (AAA) is at risk for continued enlargement after a standard aneurysm repair. Similarly, preliminary reports have shown expansion of one or both aortic necks after endovascular repair. Although some investigators have suggested that this may be a transient effect, continued dilatation at the endograft attachment site could effect the overall device stability. METHODS: As part of a multi-institutional trial of endovascular grafting for the treatment of AAA, 59 patients were successfully implanted with straight endografts between February 1993 and January 1995. A morphometric analysis of aortic neck size was undertaken with serial review of computed tomography scans available through April 1997. The neck sizes at both graft attachment sites were measured, with investigators blinded to patient identity and date of scan. Changes in minor diameter were defined, annual interval expansion rates were calculated, and the data were correlated with endoleak, device migration, aneurysm size change, endograft diameter, attachment system fractures, and initial preimplant neck size. RESULTS: Significant aortic neck enlargement, particularly at the level of the distal neck, was observed for at least 24 months after AAA repair. The annual interval dilation rates of the proximal aortic neck were 0.7 +/- 2.1 mm/year (P = .023) and 0.9 +/- 1.9 (P = .008) mm/year during the first and second years, respectively. Enlargement of the distal neck during the observation period was more marked, with corresponding annual expansion rates of 1.7 +/- 2.9 mm/year (P < .001) and 1.9 +/- 2.5 (P < .001) mm/year. In 5 patients (14%), the minor diameter of the distal neck was at least 6 mm larger than the preimplant diameter of the graft. Migration of the distal attachment system was observed in 3 of these 5 patients. Expansion rates did not have a statistically significant correlation with initial neck size, endograft dimensions, aneurysm size change, presence of endoleak, or attachment system fracture. CONCLUSIONS: Aortic neck enlargement was observed for at least 2 years after endovascular grafting. Close patient follow-up remains mandatory in lieu of the potential risk of late failure as a result of continued aortic expansion. The relative contribution of device design to this phenomenon will need to be defined. PMID- 9737452 TI - Suprarenal filter placement. AB - PURPOSE: This study was undertaken to determine the clinical outcomes for patients with Greenfield filters placed in the suprarenal (SR) inferior vena cava (IVC). METHODS: We collected data prospectively from annual follow-up evaluations of patients with filters. Patients underwent venous color-flow duplex examinations of the IVC and lower extremities, abdominal radiographs, and physical assessment. The outcomes for those patients with filters in the SR IVC were compared with the outcomes previously reported and with the outcomes for patients with filters in the infrarenal cava. RESULTS: SR placement accounted for 7.6% (148/1932) of all filter placements. Follow-up data were available for 73 placements, or 49%. No cases of renal dysfunction were related to filter placement. The rate of recurrent pulmonary embolism (PE) was 8%, and the rate of long-term caval occlusion was 2.7%. These rates did not differ statistically from the rates for patients with infrarenal filters (P > .05). Male patients tended to be older by 15 years, to have more recurrent PE, and to experience more filter migration (6 vs 2 mm). Failure of SR filters to prevent PE was associated statistically with the primary indication for placement. Recurrent PE was the indication in 5 of 6 patients who sustained PE after SR filter placement (P = .007). Filter limb fracture was seen only with the stainless-steel Greenfield filter. CONCLUSION: Greenfield filters placed above the renal vein provide protection from PE with a minimal risk of occlusion. Twenty-five years of experience with Greenfield filters shows that they are safe and effective both in young female patients of child-bearing potential and in all patients with appropriate indications for SR placement. PMID- 9737453 TI - The incidence of heparin-induced antibodies in patients undergoing vascular surgery: a prospective study. AB - PURPOSE: This study prospectively assessed the incidence of heparin-induced antibodies in patients undergoing peripheral vascular surgery and determined whether the incidence is influenced by previous heparin exposure. METHODS: Fifty four hospitalized patients (36 men and 18 women) undergoing peripheral vascular surgery and receiving intraoperative heparin anticoagulation were studied. Unfractionated porcine heparin was given for intraoperative anticoagulation and was not continued postoperatively. Carotid endarterectomy was performed in 36 patients, aortic reconstruction in 11 patients, and infrainguinal bypass in 7 patients. Plasma was tested before and after (14 +/- 7.5 [SD] days) surgery for IgG antibodies to the complex of heparin/platelet factor 4, using a standardized, validated enzyme-linked immunosorbent assay (ELISA). Results are expressed as an optical density ratio (ODR) of patient plasma to normal plasma, with the threshold for a positive result of > or = 1.8. Platelet counts and clinical outcomes were also assessed. RESULTS: The mean patient age was 67.2 +/- 9.7 years. A prior exposure to heparin was documented in 41% of patients. The mean intraoperative heparin dose was 9089 +/- 3607 units. Only 1 patient converted from a negative antibody status to a positive status (1.9%, 95% CI = 0.10% 11.18%). The change in the ELISA ODR after surgery was not significantly different for patients with (+0.042 +/- 0.272) and without (-0.022 +/- 0.299, P = 0.57) prior heparin exposure. Postoperatively, the platelet counts dropped from 227,620 +/- 78,308 microL, to 185,706 +/- 80,842 microL (P < .001). The decrease in platelet count was the same in patients with prior heparin exposure (-23.0 +/- 18.0%) and without (-18.0 +/- 14.0%, P = .46). One thrombotic complication occurred, a femorotibial bypass graft occlusion in a patient who tested negative for antibodies. CONCLUSION: Heparin-induced antibodies occur infrequently after peripheral vascular surgery. The commonly observed, mild degree of postoperative thrombocytopenia does not appear to be caused by heparin-induced antibodies. These results indicate that a standard dose of heparin for intraoperative anticoagulation during vascular surgery is not associated with a significant risk of heparin-induced thrombocytopenia and thrombosis. PMID- 9737454 TI - Warfarin improves the outcome of infrainguinal vein bypass grafting at high risk for failure. AB - OBJECTIVE: Patients with marginal venous conduit, poor arterial runoff, and prior failed bypass grafts are at high risk for infrainguinal graft occlusion and limb loss. We sought to evaluate the effects of anticoagulation therapy after autogenous vein infrainguinal revascularization on duration of patency, limb salvage rates, and complication rates in this subset of patients. METHODS: This randomized prospective trial was performed in a university tertiary care hospital and in a Veterans Affairs Hospital. Fifty-six patients who were at high risk for graft failure were randomized to receive aspirin (24 patients, 27 bypass grafts) or aspirin and warfarin (WAR; 32 patients, 37 bypass grafts). All patients received 325 mg of aspirin each day, and the patients who were randomized to warfarin underwent anticoagulation therapy with heparin immediately after surgery and then were started on warfarin therapy to maintain an international normalized ratio between 2 and 3. Perioperative blood transfusions and complications were compared with the Student t test or with the chi2 test. Graft patency rates, limb salvage rates, and survival rates were compared with the Kaplan-Meier method and the log-rank test. RESULTS: Sixty-one of the 64 bypass grafts were performed for rest pain or tissue loss, and 3 were performed for short-distance claudication. There were no differences between the groups in ages, indications, bypass graft types, risk classifications (ie, conduit, runoff, or graft failure), or comorbid conditions (except diabetes mellitus). The cumulative 5-year survival rate was similar between the groups. The incidence rate of postoperative hematoma (32% vs 3.7%; P = .004) was greater in the WAR group, but no differences were seen between the WAR group and the aspirin group in the number of packed red blood cells transfused, in the incidence rate of overall nonhemorrhagic wound complications, or in the overall complication rate (62% vs 52%). The immediate postoperative primary graft patency rates (97.3% vs 85.2%) and limb salvage rates (100% vs 88.9%) were higher in the WAR group as compared with the aspirin group. Furthermore, the cumulative 3-year primary, primary assisted, and secondary patency rates were significantly greater in the WAR group versus the aspirin group (74% vs 51%, P = .04; 77% vs 56%, P = .05; 81% vs 56%, P = .02) and cumulative limb salvage rates were higher in the WAR group (81% vs 31%, P = .01). CONCLUSIONS: Perioperative anticoagulation therapy with heparin increases the incidence rate of wound hematomas, but long-term anticoagulation therapy with warfarin improves the patency rate of autogenous vein infrainguinal bypass grafts and the limb salvage rate for patients at high risk for graft failure. PMID- 9737455 TI - Spontaneous popliteal vascular injury in the morbidly obese. AB - PURPOSE: Morbidly obese patients who sustained popliteal vascular injury after spontaneous knee dislocation (KD) were studied. METHODS: Seven morbidly obese patients (body mass index [BMI] >35 kg/m2 and >100 lb over ideal body weight) who sustained spontaneous KD while upright were reviewed. RESULTS: Severe popliteal arterial injury accompanied all spontaneous KD. The mean age of patients was 34.1 +/- 6.7 years; the mean weight was 354 +/- 150 lb (range, 220-702 lb); and mean BMI was 53 +/- 21 kg/m2 (range, 37-98.4). All had arterial avulsion and thrombosis. Three had concomitant venous injury. All underwent operative repair. Morbid obesity presented unique challenges to surgical management. Limited positioning, specialized operative tables, large incisions, deep exposure, special retraction, long operative times (mean, 537 +/- 134 minutes), and major blood loss (mean, 2.5 +/- 3 L) were standard. Five arterial injuries were repaired with interposition vein grafts, and 2 required tibial bypass. Venous repairs included thrombectomy and primary repair (n = 2) and interposition grafting (n = 1). Many complications were related to morbid obesity, including deep wound infection (n = 3), diabetic ketoacidosis (n = 2), and cor pulmonale from sleep apnea (n = 1). Despite patent grafts in all patients, 2 above-knee amputations were required for extensive neuromuscular loss. CONCLUSION: Morbid obesity is a specific risk factor for spontaneous KD and vascular injury. In addition, morbid obesity presents unique challenges to operative repair and predisposes patients to unusual major postoperative complications. PMID- 9737456 TI - A three-year follow-up on standard versus thin wall ePTFE grafts for hemodialysis. AB - PURPOSE: Expanded polytetraflouroethylene (ePTFE) grafts are the most popular prosthetic grafts for hemodialysis patients in whom autogenous fistulas cannot be constructed. Long-term studies to study the durability and complication rate of the different wall configurations of ePTFE grafts have not been carried out. The primary, secondary, and cumulative patency and other complications between standard thickness (STD) and thin wall (THN) 6 mm stretch ePTFE grafts (WL Gore & Assoc, Flagstaff, AZ) was prospectively evaluated. METHODS: From September 1993 to August 1995, 108 patients receiving new grafts were randomized into 2 groups: those receiving STD grafts (n = 56) or those receiving THN (n = 52) grafts. Data prospectively collected included day of first access, primary patency, interventions required, and long-term results. Infections, pseudoaneurysms, and mortality were also documented. Student's unpaired t-test was used to compare the 2 groups, and log-rank life tables were constructed and compared. RESULTS: Mean follow-up examination time was 38.1 +/- 0.8 months for STD grafts and 35.1 +/- 1.0 months for THN grafts (P<.03). Longer patency was noted in the STD group of grafts (18.2 months for STD vs. 12.1 months for THN). Biographical data and complications, including pseudoaneurysm (6% vs. 5%), infection (2% vs. 3%), and mortality (22% vs. 19%), between STD and THN groups were not different statistically. Mean primary (18.2 months vs. 12.1 months), secondary (20.9 months vs. 13.7 months), and cumulative patency times (22.2 months vs. 15.2 months) for the STD group were significantly more than those for the THN group (P<.000 by log rank of life tables). Other complications were not different between groups. CONCLUSION: Standard thickness ePTFE is the graft of choice when placing ePTFE arteriovenous grafts for hemodialysis. PMID- 9737458 TI - Duplex ultrasound scanning defines operative strategies for patients with limb threatening ischemia. AB - PURPOSE: To characterize the accuracy of color-flow duplex ultrasound (DUS) in planning lower extremity revascularization procedures, we prospectively compared operations predicted by means of DUS arterial scanning (DUSAS) and operations predicted by means of conventional angiography (CA) with actual operations performed in 36 patients undergoing 40 vascular reconstructions for critical (grade II/III) lower extremity ischemia. METHODS: All patients were examined with lower extremity DUSAS followed by CA. DUSAS was performed from the aorta to the pedal vessels of the affected extremity. Adequacy of inflow was assessed, and the best distal target vessel with continuous, unobstructed flow was defined. An operative prediction was made and recorded based upon the DUSAS findings, and in a blinded fashion, based upon subsequent CA. The McNemar test for comparing correlated proportions was applied to test for the statistical significance of the difference (P < .05) between correct operations predicted by DUSAS and CA. RESULTS: Of the actual operations performed, 83% were correctly predicted by means of DUSAS (95% CI; range, 77% to 89%). Seven operations were incorrectly predicted with DUSAS. Of the actual operations performed, 90% were correctly predicted by means of CA (95% CI; range, 81% to 99%). Four operations were incorrectly predicted with CA. The McNemar test determined that the difference between correct operations predicted by means of DUSAS and correct operations predicted by means of CA was not statistically significant (P = .50). CONCLUSIONS: With few exceptions, DUSAS can be used to reliably predict infrainguinal reconstruction strategies. Vessels defined as adequate with DUSAS are rarely unfit for bypass. Prospective investigation of lower extremity revascularization based solely upon DUSAS is warranted. PMID- 9737457 TI - Can intrarenal duplex waveform analysis predict successful renal artery revascularization? AB - PURPOSE: No currently available noninvasive test can preoperatively predict a successful outcome to renal revascularization. Resistance measurements from the renal parenchyma obtained with duplex sonography reflect the magnitude of intraparenchymal disease, and patients with extensive intrarenal disease may respond less favorably to revascularization. To address this question, we reviewed our (primarily) operative experience in patients undergoing renal artery revascularization, and compared the blood pressure (BP) and renal function response with resistance measurements obtained from the kidney both before and after revascularization. METHODS: During a 56-month period, 31 consecutive renal artery revascularizations (25 surgical and 6 percutaneous angioplasties) were performed in 23 patients (21 atherosclerotic, 2 fibromuscular dysplasia). Duplex sonography was performed in each patient before and after revascularization, and parenchymal diastolic/systolic (d/s) ratios were calculated. BP and renal function response to intervention were compared with measurements of intrarenal flow patterns before and after revascularization. RESULTS: Mean parenchymal peak systolic velocity was significantly higher after repair in all patients (pre repair: 19.5 +/- 1.3, postrepair: 27.2 +/- 1.7; P < .0001). Despite this, there were no statistical differences between preoperative and postoperative parenchymal d/s ratios. A favorable (cured or improved) BP response was seen in 81% (17 of 21) of revascularizations performed for hypertension. Among these successes, parenchymal d/s ratios were in the normal range (ie, > or = 0.30) both before and after repair (mean prerepair: 0.34 +/- 0.03, mean postrepair: 0.31 +/- 0.03; not significant). In 4 patients in which BP failed to improve after intervention, the d/s ratio was abnormal before surgery (< 0.3), and remained so after revascularization (mean preoperative d/s ratio: 0.18 +/- 0.04, mean postoperative d/s ratio: 0.11 +/- 0.04; P = .003). Mean preoperative parenchymal d/s ratios were significantly higher in all patients with a successful BP response when compared with failures (P = .048). Similarly, among patients with single artery repairs, mean preoperative d/s ratios approached significance in successes vs. failures (success: 0.40 +/- 0.03, failure: 0.21 +/- 0.03; P = .054). A decrease in serum creatinine greater than or equal to 20% was seen in 8 of 18 patients (44%) with ischemic nephropathy. These patients also had normal d/s ratios preoperatively (mean 0.39 +/- 0.04), whereas the 10 patients who failed to improve had significantly lower ratios (mean 0.24 +/- 0.03; P = .041). Kidney length did not correlate with d/s ratio. CONCLUSION: Although we do not believe that duplex sonographic measurement of intrarenal flow patterns alone is an accurate means of assessing main renal artery occlusive disease, the resistive indices seem to reflect the magnitude of intraparenchymal disease, and thus may provide important prognostic information for patients undergoing surgical revascularization. Our data suggest that a preoperative d/s ratio below 0.3 correlates with clinical failure relative to BP and renal function responses. PMID- 9737459 TI - Left iliac venous thrombosis caused by venous spur: treatment with thrombectomy and stent implantation. AB - PURPOSE: To determine the frequency of iliac venous spurs in left iliofemoral venous thrombosis and to report the results of interventional management of venous spurs after transfemoral venous thrombectomy. METHODS: From 1990 through 1996, 77 patients with acute iliac venous thrombosis (61 left and 16 right) underwent surgical treatment. Patients with malignant disease were excluded from this series. All patients had transfemoral venous thrombectomy with construction of an inguinal arteriovenous fistula and perioperative anticoagulation with heparin with a switch to warfarin sodium for at least 12 postoperative months. Immediate results of thrombectomy were documented by means of intraoperative completion venography. Arteriovenous fistulas were ligated 3 months after control arteriovenography. Since 1995 venous spurs eventually detected during thrombectomy were treated immediately by means of stent implantation. RESULTS: Among 61 patients with left-sided thrombosis, intraoperative phlebography revealed common iliac venous obstruction suggestive of venous spurs in 30 patients (49%). In 16 of 22 patients (73%) with untreated spurs, postoperative rethrombosis of the iliac vein was documented despite adequate anticoagulation. Only one of eight patients (13%) with stented spurs had reocclusion (chi2 test P < .01). CONCLUSION: Venous spurs are found among about half of patients with left sided iliac venous thrombosis. As long as the underlying venous pathologic process is left untreated, thrombectomy will not restore patency. Stent implantation is a simple and safe means to correct central venous strictures and provides excellent long-term results. PMID- 9737460 TI - In vivo analysis of antithrombotic effectiveness of recombinant hirudin on microvascular thrombus formation and recanalization. AB - PURPOSE: This study was undertaken to evaluate in vivo the effect of recombinant hirudin (r-hirudin [HBW 023]), a potent thrombin inhibitor, on the process of microvascular thrombus formation and recanalization. METHODS: Thrombosis was induced photochemically in distinct arterioles (n = 25) and venules (n = 30) of the ear of 16 hairless hr/hr mice (8 to 10 weeks old, 25 to 30 g of body weight). r-Hirudin (1 mg/kg of body weight) was administered intravenously directly before thrombus induction; saline-treated animals served as controls. Thrombus formation (i.e., first platelet deposition at the endothelial lining [FPD]; inner luminal diameter reduction to 50% [D/2]; complete vessel occlusion [CVO]), vessel recanalization, microcirculatory parameters, and leukocyte-endothelial cell interaction were analyzed by means of intravital fluorescence microscopy. RESULTS: Hirudin significantly delayed the process of thrombus formation compared with saline-treated controls in both arterioles (FPD: 381 +/- 80 vs 137 +/- 25 seconds, P < 0.05; D/2: 627 +/- 49 vs 501 +/- 71 seconds; CVO: 925 +/- 78 vs 854 +/- 60 seconds) and venules (FPD: 173 +/- 11 vs 59 +/- 4 seconds; D/2: 342 +/- 54 vs 228 +/- 27 seconds; CVO: 541 +/- 85 vs 344 +/- 43 seconds; P < 0.05). In addition, r-hirudin-treated animals showed an increased rate of vessel recanalization at 24 hours after thrombus induction (arterioles: 54% [7 of 13] vs 0% [0 of 12], P < 0.05; venules: 77% [10 of 13] vs 53% [9 of 17]), whereas microcirculatory parameters and leukocyte-endothelial cell interaction were not affected. CONCLUSION: Our data indicate that r-hirudin not only counteracts the process of thrombus formation but also promotes vessel recanalization, thus supporting its use in clinical microvascular surgery. PMID- 9737461 TI - Effects of heparin, desmopressin, and isovolemic hemodilution with dextran on thrombus formation in synthetic vessel grafts inserted into the vena cava of the rabbit. AB - The objective of this study was to investigate the effects of isovolemic hemodilution with dextran-70 on thrombus formation and blood flow in synthetic venous vessel grafts. Polytetrafluoroethylene grafts (length, 11 mm; inner diameter, 3 mm) were inserted into the vena cava of rabbits. Six groups were studied: (1) the control group; (2) animals that underwent isovolemic hemodilution with dextran-70 to a hematocrit of about 30%; (3) animals that underwent isovolemic dextran hemodilution combined with a bolus injection of heparin; (4) animals that underwent heparin treatment only; (5) animals that underwent isovolemic dextran hemodilution combined with infusion of desmopressin; and (6) animals that underwent an identical treatment to group 3 but with a 2 week, instead of a 2-day, follow-up. Vena cava blood flow was measured before and after hemodilution and graft insertion and at the termination of the experiments at 2 days (groups 1 to 5) and 2 weeks (group 6) after surgery. Graft patency and thrombus mass weight were evaluated. In the control group, most of the vessels occluded within 2 days. Hemodilution with dextran improved blood flow and reduced thrombus mass weight significantly. Desmopressin, which increases factor VIII, did not influence the effects of hemodilution with dextran, which suggests that the effects of dextran are not mediated by a reduction in the level of this coagulation factor. A single bolus dose of heparin did not reduce thrombus formation in the grafts but did potentiate the effects of isovolemic hemodilution on thrombus mass and graft blood flow. We conclude that isovolemic dextran hemodilution combined with a single bolus of heparin had beneficial long-lasting effects. The grafts in groups 3 and 6 were all patent. PMID- 9737463 TI - The effect of tumor necrosis factor binding protein and interleukin-1 receptor antagonist on the development of abdominal aortic aneurysms in a rat model. AB - PURPOSE: Tumor necrosis factor (TNF), interleukin 1 (IL-1), and matrix metalloproteases have been noted to be elevated in human abdominal aortic aneurysms (AAAs) as compared with normal and occlusive aortic disease. Because TNF and IL-1 have been shown to cause release of proteases that weaken the aortic matrix, it has been suggested that these cytokines may play a central role in the aortic dilatation process. To substantiate this hypothesis, we investigated the effects of TNF and IL-1 antagonists, tumor necrosis factor binding protein (TNF BP) and interleukin-1 receptor antagonist (IL-1RA), on the development of AAAs in a well-described rat model. METHODS: Isolated segments of infrarenal aorta of 16 rats were perfused with porcine elastase. In the treated group, eight rats were given intravenous TNF-BP prior to elastase perfusion, at 48 hours and at 96 hours. In the control group, eight rats were given only intravenous vehicle at the same time intervals. Isolated segments of infrarenal aorta of an additional 16 rats were perfused with porcine elastase in a similar fashion. In the treated group, eight rats were given intraperitoneal IL-1RA prior to celiotomy and every eight hours. In the control group, eight rats were given only intraperitoneal vehicle at the same time intervals. On the sixth postoperative day, all rats underwent celiotomy and measurement of the infrarenal aortic diameter with a micrometer while the animal was alive. Aortic specimens were collected on day six for hematoxylin and eosin staining, trichrome staining, and gel polyacrylamide gel electrophoresis (PAGE) zymography. RESULTS: TNF-BP was completely able to block post elastase dilation, whereas IL-1Ra seemed to have no effect. Hematoxylin and eosin staining and trichrome staining revealed that animals treated with TNF-BP had less of an inflammatory response and preservation of the elastin and smooth muscles in the media of the aortic wall as compared with animals treated with IL-1RA or vehicle. Zymography was not able to detect significant protease activity in the aortic wall of any of the rats at six days. CONCLUSION: TNF-BP, but not IL-1RA, may inhibit the development of AAAs in this model. PMID- 9737462 TI - Is smooth muscle growth in primate arteries regulated by endothelial nitric oxide synthase? AB - PURPOSE: We investigated whether control of constitutive endothelial cell nitric oxide synthase (cNOS) and nitric oxide (NO) by changes in shear stress might be important for the regulation of smooth muscle cell (SMC) growth and vascular diameter. METHODS: Bilateral femoral arteriovenous fistulas were placed in baboons to increase the blood flow in the external iliac arteries. At 2 months, the fistula was ligated on one side to restore normal flow (flow switch). RESULTS: In response to flow switch and a decrease in shear stress, iliac artery lumenal area decreased and SMC proliferation was induced. A decline in NO production, cNOS messenger RNA (mRNA), and protein were associated with these biological effects. In a subset of animals with iliac arteries under high flow, infusion of N(omega)-nitro-L-arginine, an inhibitor of cNOS, did not induce proliferation. CONCLUSION: Shear stress can regulate cNOS, vasoconstriction, and SMC proliferation. A decrease in nitric oxide may be necessary, but is not sufficient to induce SMC proliferation in response to a decrease in blood flow. PMID- 9737464 TI - Lipid uptake in expanded polytetrafluoroethylene vascular grafts. AB - PURPOSE: The mechanisms of vascular prosthesis failure are reported to be associated, in part, with an atherosclerotic degenerative process that is related to an abnormal lipid infiltration. The lipid uptake in expanded polytetrafluoroethylene (ePTFE) vascular grafts was reproduced in vitro, and the effect of time on the permeability of these prostheses was studied. METHODS: Water permeability tests were carried out under dynamic flow conditions at various hydrostatic pressures. Lipid uptake was simulated by circulating a phosphatidylcholine suspension inside an expanded Teflon prosthesis under pulsatile or continuous transmural pressure ranging between 80 mm Hg and 180 mm Hg, at a flow rate of 500 mL/min and 2000 mL/min, for a duration ranging from 2 hours to 1 month. RESULTS: Water permeability tests indicated that under hydrostatic pressures of 180 mm Hg and 300 mm Hg, water percolated through the prosthesis wall after an exposure of 720 minutes and 75 minutes, respectively. After exposing the prostheses to the lipid dispersion under the various flow conditions, the fluid convection through the wall occurred. Preferential convection pathways with a constant periodicity were observed across the length of each prosthesis and were, therefore, associated with regularly spaced perforations depicted in the structure of the devices. Phospholipids gradually agglomerated within the prosthesis wall, allowing a restrictive molecular mobility. Infrared spectroscopy results indicated that the lipid uptake depended on the transmural pressure and time of exposure. CONCLUSION: The occurrence of the membrane permeability may be associated with the dilatation and plastic deformation of the prosthesis. Lipid uptake occurs in ePTFE grafts after an aggressive kinetic process. PMID- 9737465 TI - Increased plasma vascular endothelial growth factor among patients with chronic venous disease. AB - Skin damage in the presence of chronic venous disease is partially mediated through leukocytes. The endothelium is activated and exhibits proliferation in the skin. Up-regulation of vascular endothelial growth factor (VEGF) expression in the skin of patients with chronic venous disease has been demonstrated with immunohistologic techniques. Abnormal VEGF expression can have local deleterious effects. The aim of this study was to determine whether patients with chronic venous disease have elevated plasma levels of VEGF. We conducted a prospective study with 30 patients with varicose veins of clinical, etiologic, anatomic, and pathologic class C3 (normal skin, n = 15) and C4 (trophic skin changes, n = 15) and 25 control subjects with no clinical evidence of venous or arterial disease of the lower limb. Blood samples were collected from a foot vein of each subject before and after a period of experimental venous hypertension produced by means of standing. Assay of VEGF protein was performed with a sandwich enzyme-linked immunosorbent assay. Plasma VEGF level was elevated in both groups of patients with venous disease compared with the control group. The median VEGF levels among patients were 81 pg/mL (interquartile range [IQR] 56 to 122) supine and 98 pg/mL (IQR 63 to 153) after standing for 30 minutes. Median VEGF levels among control subjects were 52 pg/mL (IQR 35 to 71) lying supine and 60 pg/mL (IQR 39 to 105) after standing for 30 minutes. Experimental venous hypertension caused a small rise in VEGF levels among the patients but not the control subjects. Further studies are required to determine whether increased VEGF expression contributes to tissue injury in chronic venous disease. PMID- 9737466 TI - Endovascular retrieval of two migrated venous stents by means of balloon catheters. AB - The usefulness of vascular stenting was demonstrated in both arterial and venous applications to restore patency and improve suboptimal results after percutaneous transluminal angioplasty. Dislodgment of venous stents with an embolization into the right cavities or the pulmonary artery, however, is one of the most feared complications of this procedure. Percutaneous removal of these migrated stents is an appealing method of replacing more invasive operative intervention with cardiopulmonary bypass, which may be very hazardous in these often severely ill patients. We describe the cases of two patients with stents that migrated into the right ventricle and the pulmonary artery. In one patient, we were able to successfully remove these stents by using an angioplasty balloon with an operative extraction from the iliac vein, thereby obviating the need for a major operative procedure. PMID- 9737467 TI - Mycotic renal artery degeneration and systemic sepsis caused by infected renal artery stent. AB - A case of Staphylococcus aureus renal artery stent infection was studied. Fourteen days after the procedure, the patient had a fever, hypotension, and an elevated white blood cell (WBC) count. Blood cultures were positive for S. aureus on admission and during the patient's hospitalization, despite intravenous vancomycin therapy. Evaluation included serial CT scans, revealing increasing persistent inflammation with development of multiple renal intraparenchymal abscesses, and arteriography, showing marked degeneration of the renal artery. Therapy required resection of the renal artery/stent and nephrectomy. This case confirms the severe nature of S. aureus stent infection; we recommend prophylactic antibiotics before these procedures, as well as expeditious evaluation and consideration for aggressive surgical therapy if this complication is suspected. PMID- 9737468 TI - The management of massive ultrafiltration distending the aneurysm sac after abdominal aortic aneurysm repair with a polytetrafluoroethylene aortobiiliac graft. AB - Collections of serous fluid surrounding prosthetic grafts can be caused by infection or transudation of serum, and making the distinction is often troublesome. Bergamini and his colleagues developed a dog model of low-grade prosthetic graft contamination with Staphylococcus epidermatis. All animals developed evidence of graft infection, and 13 of 18 dogs developed a fluid-filled perigraft cyst. Signs of systemic infection, however, were present in only 1 animal, and the Staphylococcus epidermatis study strain was isolated from the tissue surrounding the graft in only 1 dog. The authors had to disrupt the biofilm to achieve positive cultures in 14 of 18 animals. This animal model seemed to conform to clinical experience and placed great emphasis on the role of indolent infections in the pathogenesis of perigraft fluid collection. It is equally clear that perigraft fluid collections may result from transudation of fluid through the prosthetic surfaces, which act similar to a dialysis membrane under certain circumstances. Noninfectious seromas are characterized generally by the accumulation of clear serous fluid with a protein and glucose content of serum and the lack of acute inflammatory cells when the sediment is examined. The need to distinguish between these 2 forms of fluid accumulation became important in the treatment of a 62-year-old man who was seen 2 1/2 years after the repair of an abdominal aortic aneurysm with an aortobiiliac stretch polytetrafluoroethylene (PTFE) prosthesis. There was no evidence of infection, and there was a 12 cm cystic mass surrounding a patent PTFE prosthesis. PMID- 9737469 TI - Superior vena cava syndrome caused by chronic hemodialysis catheters: autologous reconstruction with a pericardial tube graft. AB - Superior vena cava (SVC) syndrome caused by long-term use of indwelling catheters is an infrequent but increasingly common complication. Because collateralization often is sufficient, surgical treatment rarely is indicated. We present a case of a patient with severe symptomatic SVC syndrome as a result of the long-term use of indwelling hemodialysis catheters. The SVC was reconstructed with a pericardial tube graft. Magnetic resonance angiography performed 13 months after the operation showed patency of the graft. The patient continues to be free of symptoms. PMID- 9737470 TI - Concurrence of anaphylaxis and acute heparin-induced thrombocytopenia in a patient with heparin-induced antibodies. AB - We report the occurrence of acute heparin-induced thrombocytopenia in a patient with anaphylaxis that began immediately after an intravenous bolus dose of unfractionated heparin. This case report is the first to document the concurrence of these 2 reactions to heparin. An abrupt fall in platelet count was documented immediately after the anaphylactic response. Study results for antibodies characteristic of heparin-induced thrombocytopenia were positive in 2 assays: serotonin release assay and heparin platelet factor 4 enzyme-linked immunosorbent assay. The patient's antibody was exclusively immunoglobulin G. Any explanation for the relationship between the antibody response observed and the histamine release remains speculative. PMID- 9737471 TI - Inferior epigastric artery pseudoaneurysm: a complication of paracentesis. AB - Two patients had inferior epigastric artery pseudoaneurysms after therapeutic paracentesis for ascites caused by portal hypertension. The first patient, a 62 year-old man, had a two-week history of left lower quadrant pain, tenderness, and nonpulsatile mass after a paracentesis for ascites. A left inferior epigastric artery pseudoaneurysm measuring 10 cm in diameter and 20 cm in length was diagnosed by means of Duplex ultrasound and arteriography. The patient was treated with percutaneous embolization, with successful thrombosis of the pseudoaneurysm. The second patient, a 33-year-old woman, had a six-week history of left lower quadrant pain, tenderness, and nonpulsatile mass after a paracentesis for ascites. Computerized tomography and arteriography showed a left inferior epigastric artery pseudoaneurysm, measuring 7 cm in diameter and 9 cm in length. The patient was treated with percutaneous embolization with successful thrombosis of the pseudoaneurysm. Both patients were discharged in good condition 2 days after embolization. Inferior epigastric artery pseudoaneurysm is a complication of paracentesis, and percutaneous embolization may be preferable to surgical repair in patients with chronic liver failure and portal hypertension. PMID- 9737472 TI - Regarding "Importance of intercostal artery reattachment during thoracoabdominal aortic aneurysm repair". PMID- 9737473 TI - Low-molecular-weight heparins: pharmacologic profile and product differentiation. AB - The interchangeability of low-molecular-weight heparins (LMWHs) has been the subject of discussion since these products were first introduced for the prophylaxis of deep vein thrombosis. Experimental evidence now exists to show that LMWHs differ from each other in a number of characteristics. Products have been differentiated on the basis of molecular weight and biologic properties, but only limited information derived from the clinical setting is available. Potency has been described on the basis of anti-Factor Xa activity, but at equivalent anti-Xa activities, the anti-Factor IIa activity of different products shows marked variations. At the relatively small doses used for the management of postsurgical deep vein thrombosis, the effect of these interproduct differences may be relatively minor, but as LMWHs are developed for therapeutic use at much higher doses, such differences may become clinically important. Variations in safety and efficacy reported in clinical trials of LMWHs may reflect the known differences in their molecular composition and pharmacologic properties. PMID- 9737474 TI - Clinical potential of antithrombotic drugs in coronary syndromes. AB - Thrombosis is responsible for most of the acute manifestations of coronary artery disease, including unstable angina and non-Q-wave myocardial infarction. Antithrombotic therapy with antiplatelet agents and anticoagulants plays a major role in decreasing the risk of ischemic events in such patients. As thrombin generation plays a key role in the pathogenesis of thrombosis, recent studies have focused on thrombin inhibition in the management of acute ischemia. Heparin is the most widely used anticoagulant in the acute phase. Heparin given in therapeutic doses intravenously has been shown to be more effective than aspirin in decreasing the risk of death or myocardial infarction in patients with unstable angina. Low-molecular-weight heparin (LMWH) has improved pharmacologic and pharmacokinetic properties over standard heparin and this may provide greater efficacy and safety. LMWH may be given at fixed subcutaneous dose without monitoring and, therefore, is of greater clinical utility and is more cost effective than standard heparin. Several LMWHs have been evaluated in the management of acute coronary syndromes and have shown equivalent or improved efficacy compared with standard heparin. As heparin in combination with aspirin is now the treatment of choice in acute unstable coronary syndromes, LMWH could potentially improve the outcome in such patients. PMID- 9737475 TI - Rationale for the management of coronary syndromes with low-molecular-weight heparins. AB - The well-documented disadvantages of unfractionated heparin in the management of coronary syndromes, such as unpredictable bioavailability and maintenance of therapeutic range, has prompted several studies into the benefits of low molecular-weight heparins (LMWHs). The favorable pharmacologic properties of LMWHs include a binding affinity for antithrombin III, anti-factor IIa activity, excellent bioavailability, minimal protein binding, predictable anticoagulant response, and clinical tolerance by patients. LMWHs are also characterized by having a specific anti-factor Xa effect and inducing only a small prolongation in general clotting tests-i.e., activated partial thromboplastin time, prothrombin time, and antithrombin activity-when used in high doses. Several studies have recently demonstrated that LMWHs are superior to placebo and are at least equal or superior to standard heparin when added to aspirin for the treatment of unstable angina and following non-Q-wave myocardial infarction. These studies, which include Thrombolysis in Myocardial Infarction (TIMI) 11A and Efficacy and Safety of Subcutaneous Enoxaparin versus intravenous unfractionated heparin in Non-Q-wave Coronary Events (ESSENCE), will be reviewed and discussed. PMID- 9737476 TI - Low-molecular-weight heparins in non-ST-segment elevation ischemia: the ESSENCE trial. Efficacy and Safety of Subcutaneous Enoxaparin versus intravenous unfractionated heparin, in non-Q-wave Coronary Events. AB - Combination antithrombotic therapy with heparin plus aspirin decreases the risk of recurrent ischemic events in patients with acute coronary syndromes without persistent ST-segment elevation. Compared with standard unfractionated heparin, low-molecular-weight heparin (LMWH) has a more predictable antithrombotic effect, is easier to administer, and does not require coagulation monitoring. At 176 hospitals in 3 continents, 3,171 patients with rest unstable angina or non-wave myocardial infarction were randomly assigned to either enoxaparin (a LMWH), 1 mg/kg twice daily subcutaneously, or to continuous intravenous unfractionated heparin, for a minimum of 48 hours to a maximum of 8 days. Trial medication was administered in a double-blind, placebo-controlled fashion. At 14 days, the primary endpoint, the composite risk of death, myocardial infarction, or recurrent angina with electrocardiographic changes or prompting intervention, was significantly lower in patients assigned to enoxaparin compared with heparin (16.6% vs 19.8%; odds ratio [OR] 1.24; 95% confidence interval [CI] 1.04-1.49; p = 0.019). At 30 days, the composite risk of death, myocardial infarction, or recurrent angina remained significantly lower in the enoxaparin group compared with the unfractionated heparin group (19.8% vs 23.3%, OR 1.23; 95% CI 1.0-1.46, p = 0.016). The rate of revascularization procedures at 30 days was also significantly lower in patients assigned to enoxaparin (27.1% vs 32.2%, p = 0.001). The 30-day incidence of major bleeding complication was 6.5% versus 7.0% (p = not significant), but the incidence of minor bleeding was significantly higher in the enoxaparin group (13.8% vs 8.8%, p <0.001) due primarily to injection-site ecchymosis. Thus, combination antithrombotic therapy with enoxaparin plus aspirin is more effective than unfractionated heparin plus aspirin in decreasing ischemic outcomes in patients with unstable angina or non-Q wave myocardial infarction in the early (30 days) phase. The lower recurrent ischemic event rate seen with the LMWH, enoxaparin, is achieved without an increase in major bleeding, but with an increase in minor bleeding complications due mainly to injection-site ecchymosis. PMID- 9737477 TI - Antithrombotics in interventional cardiology: optimizing treatment and strategies. AB - Although the use of coronary stents has virtually abolished the threat of periprocedural obstructive dissection, subacute and acute intracoronary thrombosis and late restenosis remain a major problem with catheter-guided transluminal coronary interventions, despite significant technical advances over the last 10 years. Acute stent thrombosis emerged as a new problem with the introduction of metallic coronary prostheses (stents), which unfortunately represent an ideal stimulus for platelet deposition. Recently, dramatic progress has been achieved by focusing on the inhibition of thrombin and platelets, before and during interventional procedures. This has stimulated the search for powerful and well-tolerated antithrombotic agents-platelet inhibitors and antithrombins-so that long-term (oral) administration may become possible, if necessary. The current roles of unfractionated and low-molecular-weight heparins (LMWHs), direct thrombin inhibitors (such as hirulog and hirudin), antiplatelet agents (such as aspirin, clopidogrel, and ticlopidine) and the potential of the glycoprotein IIb/IIa receptor blockers are reviewed and put into perspective with respect to their acute and long-term clinical value. PMID- 9737478 TI - Low-molecular-weight heparins in coronary stenting (the ENTICES trial). ENoxaparin and TIClopidine after Elective Stenting. AB - The role of low-molecular-weight heparins (LMWHs) in the management of stent thrombosis, although expected to produce fewer hemorrhagic complications than warfarin anticoagulation regimens, is poorly defined. The ENoxaparin and TIClopidine after Elective Stenting (ENTICES) trial was designed to compare a combination of a LMWH (enoxaparin), ticlopidine, and aspirin with the conventional warfarin anticoagulant treatment in patients who received coronary stents, in an effort to decrease stent thrombosis and ischemic clinical events. The results show that the enoxaparin regimen produced significantly fewer clinical events and vascular complications than the conventional warfarin anticoagulant regimen. PMID- 9737479 TI - Monitoring of low-molecular-weight heparins in cardiovascular disease. AB - Thrombin generation is a key event in the pathophysiology of coronary syndromes and provides the rationale for treatment with anticoagulants. Unlike standard heparin, low-molecular-weight heparin (LMWH) has little effect on activated partial thromboplastin time. LMWH treatment has been monitored by measurement of anti-Factor Xa activity, but this may not accurately reflect the anticoagulant action because LMWHs also inhibit Factor II. The Heptest is a clotting assay that is sensitive to both anti-Xa and anti-IIa activity, as well as inhibition of the extrinsic pathway by LMWH-stimulated release of tissue factor pathway inhibitor. The plasma thrombin neutralization assay has also been used to measure LMWH and to detect low concentrations to which chromogenic assays are insensitive. In the clinical setting, monitoring the anti-Xa activity in patients treated with LMWH after acute deep vein thrombosis offered no advantages over a standard weight adjusted dose. Moreover, in acute coronary syndromes there is no increase in major hemorrhage rates with weight-adjusted LMWH. Monitoring of LMWH concentrations may be advisable in the presence of comorbid conditions carrying an increased risk of hemorrhage, such as renal disease, advanced age, severe over or underweight, or a history of previous bleeding episodes. PMID- 9737480 TI - Metabolic derangement in ischemic heart disease and its therapeutic control. AB - The term myocardial ischemia describes a condition that exists when fractional uptake of oxygen in the heart is not sufficient to maintain the rate of cellular oxidation. This leads to extremely complex situations that have been extensively studied in recent years. Experimental research has been directed toward establishing the precise sequence of biochemical events leading to myocyte necrosis, as such knowledge could lead to rational treatments designed to delay myocardial cell death. At the present time, there is no simple answer to the question of what determines cell death and the failure to recover cell function after reperfusion. Problems arise because: (1) ischemic damage is not homogeneous and many factors may combine to cause cell death; (2) severity of biochemical changes and development of necrosis are usually linked (both the processes being dependent on the duration of ischemia) and it is impossible to establish a causal relation; and (3) the inevitability of necrosis can only be assessed by reperfusion of the ischemic myocardium. Restoration of flow, however, might result in numerous other negative consequences, thus directly influencing the degree of recovery. From the clinical point of view, we have recently learned that there are several potential manifestations and outcomes associated with myocardial ischemia and reperfusion. Without a doubt, ventricular dysfunction (either systolic or diastolic) of the ischemic zone is the most reliable clinical sign of ischemia, since electrocardiographic changes and symptoms are often absent. The ischemia-induced ventricular dysfunction, at least initially, is reversible, as early reperfusion of the myocardium results in restoration of normal metabolism and contraction. In the ischemic zone, recovery of contraction may occur instantaneously or, more frequently, with a considerable delay, thus yielding the condition recently recognized as the "stunned" myocardium. On the other hand, when ischemia is severe and prolonged, cell death may occur. Reperfusion at this stage is associated with the release of intracellular enzymes, damage of cell membranes, influx of calcium, persistent reduction of contractility, and eventual necrosis of at least a portion of the tissue. This entity has been called "reperfusion damage" by those who believe that much of the injury is the consequence of events occurring at the moment of reperfusion rather than a result of changes occurring during the period of ischemia. The existence of reperfusion damage, however, has been questioned, and it has been argued that, with the exception of induction of arrhythmias, it is difficult to be certain that reperfusion causes further injury. The existence of such an entity has clinical relevance, as it would imply the possibility of improving recovery with specific interventions applied at the time of reperfusion. In 1985, Rahimtoola described another possible outcome of myocardial ischemia. He demonstrated that late reperfusion (after months or even years) of an ischemic area showing ventricular wall-motion abnormalities might restore normal metabolism and function. He was the first to introduce the term "hibernating myocardium," referring to ischemic myocardium wherein the myocytes remain viable but in which contraction is chronically depressed. In this article we review our data on metabolic changes occurring during ischemia followed by reperfusion, obtained either in the isolated and perfused rabbit hearts or in ischemic heart disease patients undergoing intracoronary thrombolysis or aortocoronary bypass grafting. PMID- 9737481 TI - Treating ischemic heart disease by pharmacologically improving cardiac energy metabolism. AB - An increasing number of clinical and experimental studies have shown that optimizing energy metabolism in the heart is an effective approach to decreasing the symptoms associated with myocardial ischemia. In particular, increasing myocardial glucose metabolism can benefit heart function and/or lessen tissue injury. However, high levels of circulating fatty acids will markedly decrease glucose metabolism in the heart. These high levels of fatty acids occur in most clinically relevant conditions of myocardial ischemia, and can contribute to the severity of ischemic injury. A number of pharmacologic agents are now available that directly stimulate myocardial glucose metabolism or indirectly stimulate glucose metabolism secondary to an inhibition of fatty acid metabolism. One agent that appears to act by this mechanism is trimetazidine, which we recently found stimulates glucose oxidation in isolated rat hearts perfused with high levels of fatty acids. Clinical studies have also shown that trimetazidine has cardioprotective effects in the setting of myocardial ischemia. As a result, optimizing energy metabolism with agents such as trimetazidine may have considerable promise as a new approach to treating cardiovascular disease. PMID- 9737482 TI - Is the cytoprotective effect of trimetazidine associated with lipid metabolism? AB - Trimetazidine is an anti-ischemic compound devoid of hemodynamic effect, which was recently suspected to induce cardioprotection at the cellular level by a mechanism involving lipid metabolism. The effect on trimetazidine was evaluated in vivo by determination of rat cardiac fatty acid composition, and in vitro by investigation of the phospholipid metabolism in cultured rat cardiomyocytes. In rats, a 4-week trimetazidine treatment induced a significant decrease in the phospholipid content in linoleic acid, balanced by a small increase in oleic and stearic acids. These changes were not correlated with similar alterations in plasma fatty acid composition. In isolated cells, the time-dependent incorporation of labeled precursors of membrane phospholipid ([3H]inositol, [14C]ethanolamine, [14C]choline, [3H]glycerol, [14C]arachidonic acid, and [14C]linoleic acid 10 micromol/L) was compared in trimetazidine-treated cells and control cells. In trimetazidine-treated cells, arachidonic acid incorporation was increased in the phospholipid, but not in other lipid fractions. This enhanced fatty acid utilization elicited a net increase in the total arachidonic acid uptake. The incorporation of [14C] inositol in phosphatidylinositol was strongly stimulated by trimetazidine, although the uptake of inositol was not altered. The difference was significant within 30 minutes, and reached +70%(in trimetazidine treated cells) after 150 minutes. A similar result was obtained with ethanolamine as phosphatidylethanolamine precursor, where turnover increased by 50% in trimetazidine-treated cells. Conversely, the incorporation of choline in phosphatidylcholine was not significantly affected by the presence of trimetazidine. In conclusion, trimetazidine appears to interfere with the metabolism of phospholipids in cardiac myocytes in a manner that could indicate an increased phosphatidylinositol turnover and a redirection of cytidine triphosphate (CTP) utilization toward phosphatidylethanolamine instead of phosphatidylcholine turnover. This overall phospholipid turnover increase may contribute to a reorganization of the fatty acid utilization balance in the heart, which could lead to a lowered availability of fatty acids for energy production. PMID- 9737483 TI - Ionic and metabolic imbalance as potential factors of ischemia reperfusion injury. AB - This study examined the influence of metabolic substrates on the effects of trimetazidine on functional and metabolic aspects of the ischemic reperfused heart. Isovolumic rat hearts were submitted to a 30-minute period of global mild ischemia (coronary flow decreased by an average of 70%) and then reperfused at constant preischemic coronary flow rate. Either glucose (11 mM) or glucose and palmitic acid (0.1 mM) were used as metabolic substrates. Trimetazidine (6 x 10( 7)M) markedly reduced the increase in diastolic pressure that occurred on reperfusion after the ischemic episode, whatever the exogenous substrate used. However, in those hearts that received fatty acid, the postischemic increase in diastolic pressure was abolished. Ischemia-induced increase in acyl carnitine levels-determined as indicators of fatty acid utilization by myocardial cells-was significantly decreased by trimetazidine in those hearts receiving fatty acid. Also, similar effects to those of trimetazidine on the postischemic increase in diastolic pressure and on tissue levels of acyl carnitine were obtained in the presence of dichloroacetate. Moreover, the presence of trimetazidine was associated with a reduction in the intracellular pH decrease during ischemia in those hearts receiving fatty acid. Combined with previous studies, these results suggest that an improved metabolic balance by trimetazidine may well consequently decrease the ionic imbalance after a transient period of ischemia. PMID- 9737484 TI - Coronary artery constriction in rats: necrotic and apoptotic myocyte death. AB - The purpose of this study was to determine whether coronary artery narrowing was associated with the activation of necrotic and apoptotic myocyte cell death in the myocardium and whether these 2 forms of cell death were restricted to the left ventricle, or involved the other portions of the heart. Coronary artery narrowing was surgically induced in rats, and the animals were killed from 45 minutes to 12 days after surgery. Myocyte apoptosis was detected by the terminal deoxynucleotidyl transferase assay, confocal microscopy, and deoxyribonucleic acid (DNA) agarose gel electrophoresis. Myocyte necrosis was identified by myosin monoclonal antibody labeling of the cytoplasm. A separate group of animals was treated with trimetazidine in an attempt to interfere with tissue injury. Coronary artery narrowing was characterized by myocyte apoptosis in the left ventricle and interventricular septum, which progressively increased from 45 minutes to 6 days. However, apoptosis was not observed at 12 days. Conversely, myocyte necrosis reached its maximum value at 1 day and was still present at 12 days. This form of cell death affected not only the left ventricular free wall and interventricular septum, but also the right ventricle. Cell necrosis markedly exceeded apoptosis at all intervals. At the peak of cell death, myocyte necrosis was 52-fold and 33-fold higher than apoptosis in the left ventricle and septum. In conclusion, necrotic myocyte cell death is the prevailing form of damage produced by coronary artery narrowing, but apoptotic cell death contributes to the loss of myocytes in the ischemic heart. Trimetazidine treatment attenuated the extent of myocardial damage produced by global ischemia. PMID- 9737485 TI - Trimetazidine-induced enhancement of myocardial glucose utilization in normal and ischemic myocardial tissue: an evaluation by positron emission tomography. AB - Trimetazidine has an anti-ischemic effect in angina pectoris. This agent has no hemodynamic effects, and its benefit is presumed to be based on a metabolic mechanism of action. A group of 33 dogs undergoing openchest left anterior descending coronary artery (LAD) ligation causing prolonged ischemia were imaged with quantitative positron emission tomography (PET) using 2-[18F]fluoro-2-deoxy D-glucose (18FDG) to measure regional glucose metabolic utilization (rGMU) and [11C]acetate to measure regional monoexponential washout rate constant (Kmono) for oxidative metabolism in nonrisk and ischemic-risk myocardium. A total of 20 dogs were pretreated with trimetazidine at low dose (n = 10, 1 mg/kg) and high dose (n = 10, 5 mg/kg) and compared with 13 control dogs. Microsphere-measured myocardial blood flow (mL/min/g) was measured preocclusion and repeated hourly after occlusion and expressed as a ratio of preocclusion myocardial blood flow to verify a stable level of ischemia during PET. No differences were seen in postocclusion ischemic risk/nonrisk myocardial blood flow between treatment groups (p = not significant [NS]). Preocclusion and hourly measurements of heart rate and blood pressure corrected for baseline revealed no difference in control dogs versus trimetazidine (low-dose and high-dose) groups (p = NS). 18FDG-derived rGMU (micromol/min/g) was increased in high-dose trimetazidine versus control dogs in nonrisk and ischemic risk groups, respectively (1.16+/-0.57 vs 0.51+/ 0.38 and 0.43+/-0.29 vs 0.20+/-0.14; p <0.05). rGMU was increased proportionately in nonrisk and ischemic risk in all groups without significant differences when corrected for nonrisk rGMU (ischemic risk/nonrisk was 0.92+/-1.3 vs 0.64+/-0.66 vs 0.40+/-0.22 for control dogs, all trimetazidine and high-dose trimetazidine groups). Kmono (min(-1) was not altered in any group (nonrisk = 0.13+/-0.03 vs 0.13+/-0.03 vs 0.14+/-0.02 and ischemic risk = 0.18+/-0.05 vs 0.17+/-0.06 vs 0.16+/-0.06 for control dogs, all trimetazidine and high-dose trimetazidine groups, respectively; p = NS for nonrisk vs ischemic risk, between and within groups). Our data verify that trimetazidine does not alter hemodynamic porameters. It increases total glucose utilization (oxidative and glycolytic) in myocardium without preferential increase in ischemic tissue. Absence of change in total oxidative metabolism suggests increased glucose metabolism is predominantly glycolysis or an increase in glucose oxidation with similar decrease in fatty acid oxidation. PMID- 9737486 TI - Metabolic management of ischemic heart disease: clinical data with trimetazidine. AB - The metabolic management of heart disease represents a promising new strategy for ischemic syndromes. This review focuses on the clinical studies performed with trimetazidine, a cellular anti-ischemic agent, either as monotherapy or in combination with other drugs. Acute and chronic administration of trimetazidine significantly increased total work, exercise duration, and time to 1-mm ST segment depression, without any change in heart rate, systolic blood pressure, or rate-pressure product at the same workload, as compared with placebo. Chronic administration studies have been performed comparing the anti-ischemic effects of trimetazidine with nifedipine or propranolol in patients with chronic stable angina. In these studies, the number of anginal attacks was significantly decreased in all groups. Ergometric results showed that trimetazidine increased duration of exercise, time to 1-mm ST segment depression, time to angina, and decreased ST segment depression at peak exercise similarly to propranolol or nifedipine treatment. Rate-pressure product at the same workload remained unchanged in the patients treated with trimetazidine in comparison with baseline treatment, suggesting no effect of the drug on oxygen demand, whereas it was decreased during treatment with nifedipine and propanolol, because of the hemodynamic activity of these drugs. The therapeutic value of adding trimetazidine to either calcium antagonists or beta blockers in patients with ischemic heart disease has also been demonstrated. Unlike classic anti-ischemic agents, treatment with trimetazidine is not accompanied by any modification in hemodynamic parameters, confirming the experimental data showing a unique mechanism of action via a direct effect on the ischemic myocardium. PMID- 9737487 TI - Energy substrate metabolism, myocardial ischemia, and targets for pharmacotherapy. AB - Myocardial ischemia is essentially a metabolic event. In this review we will try to distill the essence of a complex series of molecular reactions triggered by the sudden reduction or cessation of blood flow to the heart. We recognize that it is difficult to describe even simple metabolic changes occurring in ischemia without a brief recap of pathways of energy transfer in the normal myocardium. We will therefore begin with a description of the energy substrate supply to a system that is best defined as the heart's remarkable ability for efficient conversion of chemical into mechanical energy. At the core of the system are rates of oxidative phosphorylation of adenosine diphosphate (ADP) that exactly match rates of adenosine triphosphate (ATP) hydrolysis. We will then describe the consequences of a sudden interruption to this balance, namely ischemia. At the same time we will explore metabolic strategies that may be employed to lessen the consequences of ischemia on contractile function, highlighting areas of future research and clinical investigation. The review is not meant to be comprehensive. Its main aim is to discuss the concept of pharmacotherapy as an intervention in altered cellular metabolism, akin to the concept of reperfusion therapy as an intervention in obstructed coronary arteries. PMID- 9737488 TI - Measurements of myocardial metabolism in patients with ischemic heart disease. AB - Therapeutic approaches to left ventricular dysfunction in ischemic heart disease are likely to be guided by the answers to 2 major questions: (1) which mechanisms account for the deterioration or loss of contractile function, and (2) can contractile function be improved or restored therapeutically? Both questions can face considerable diagnostic challenges. Two main mechanisms have been implicated in the pathophysiology of reversible dysfunction-myocardial hibernation and myocardial stunning. Both general concepts share a number of clinical features so that standard clinical approaches often fail to discriminate between them. The presence of regionally increased fibrosis in scar tissue formation associated with decreased blood flow further complicates the search for truly reversible dysfunction. PMID- 9737489 TI - Preliminary study of the capnogram waveform area to screen for pulmonary embolism. AB - STUDY OBJECTIVE: Acute pulmonary embolism (PE) increases alveolar dead space, which can dilute carbon dioxide content in exhaled breath. This study was undertaken to determine whether the capnogram waveform area from patients with PE is decreased compared with that from patients without PE and to examine the potential role of the capnogram waveform area as a screening test for PE. METHODS: Studies were conducted on patients from a large, urban emergency department. Steady-state capnograms were collected from subjects breathing room air, and data were electronically stored for later analysis; arterial blood was collected for blood gas analysis. PE was diagnosed or excluded on the basis of appropriate combinations of isotopic ventilation-perfusion lung scanning, lower extremity venous Doppler ultrasound studies, and pulmonary angiography. Capnogram areas were measured by tracing waveforms on printed copies with an electronic digitizing tablet. RESULTS: The mean capnogram area from patients with PE (n=19) was 28+/-10 mm Hg x sec, significantly less than for patients without PE (n=120), 53+/-17 mm Hg x sec (95% confidence interval [CI] for a difference of 25 mm Hg x sec, 17 to 33 mm Hg x sec). At a cutoff of 50 mm Hg x sec, the test sensitivity was 100% (95% CI, 82% to 100%), with a negative likelihood ratio (LR-) of .17, and the specificity was 53% (95% CI, 44% to 62%), with a positive likelihood ratio (LR+) of 2.1. At a cutoff of 25 mm Hg x sec, the test sensitivity was 42% (95% CI, 20% to 67%; LR-, .6) and the specificity was 97% (95% CI, 92% to 99%; LR+, 12.3). The area under the smoothed receiver operating characteristic curve for the capnogram area was .896 (95% Cl, .80 to .99). CONCLUSION: The capnogram waveform area may be useful in screening for PE in the ED. PMID- 9737490 TI - Worst headache and subarachnoid hemorrhage: prospective, modern computed tomography and spinal fluid analysis. AB - STUDY OBJECTIVE: This study investigated the hypothesis that modern computed tomographic (CT) imaging is sufficient to exclude subarachnoid hemorrhage (SAH) in patients with severe headache. METHODS: All 38,730 adult patients who presented to Hermann Hospital in Houston, Texas, during a 16-month period were prospectively screened to detect those with "the worst headache of my life." Two neuroradiologists blinded to the study hypothesis interpreted the CT scans. Patients with negative scans underwent comprehensive cerebrospinal fluid (CSF) analysis including cell count in first and last tubes, visual and spectrophotometric detection of xanthochromia, and CSF D-dimer assay. RESULTS: A chief complaint of headache was elicited in 455 patients, and 107 of these had "worst headache" and were enrolled in the study. CT-confirmed SAH was found in 18 of the 107 (17%). Only 2 patients (2.5%, 95% confidence interval, .3% to 8.8%) had SAH detected by CSF analysis among those with negative CT imaging result. CSF spectrophotometric detection was the most sensitive test for blood. Three patients with less than 6 red blood cells in tube 1 had positive spectrophotometric results, but in all 3, tube 4 was negative on spectrophotometric analysis, suggesting a high false-positive rate. CONCLUSION: Modern CT imaging is sufficient to exclude 97.5% of SAH in patients presenting to the ED with "worst headache" symptoms. PMID- 9737491 TI - Accuracy of reagent strips in detecting hypoglycemia in the emergency department. AB - STUDY OBJECTIVE: Although reagent strips are commonly used, their reliability to estimate blood glucose concentration and guide administration of dextrose solutions in the emergency department environment has not been proved. We determined the accuracy of visually interpreted reagent strips (Chemstrip bG, Boehringer Mannheim Corp, Indianapolis, IN) and their ability to identify hypoglycemic patients in the ED. METHODS: We conducted a prospective, nonrandomized blinded clinical study of the visual estimation of blood glucose values by ED personnel using Chemstrip bG reagent strips during a 4-month period. Simultaneously obtained blood samples sent for laboratory glucose determination served as controls. The study was conducted at a large university hospital ED with an urban patient population. A convenience sample of 215 adult ED patients underwent serum glucose determination with data form completion. No study intervention was tested, although timing of administration of dextrose solutions, if given, was recorded. RESULTS: Hypoglycemia was defined as a glucose concentration less than 60 mg/dL on standard laboratory analysis. Reagent strips identified 28 of 29 of these patients (sensitivity=97%), and 171 of 182 patients without hypoglycemia (specificity=94%, negative predictive value=99%) compared with control samples. The 1 false-negative reagent strip reading of 80 mg/dL was obtained from blood stored in a serum separator tube and had a laboratory glucose value of 39 mg/dL. Eighty-seven percent of the reagent strips were within +/-60 mg/dL of the control value for the laboratory glucose reference range less than 350 mg/dL. CONCLUSION: Visually interpreted Chemstrip bG reagent strips provide an acceptable estimation of blood glucose concentration in the ED and are highly sensitive in detecting hypoglycemia. PMID- 9737492 TI - Who reviews the reviewers? Feasibility of using a fictitious manuscript to evaluate peer reviewer performance. AB - STUDY OBJECTIVE: To determine whether a fictitious manuscript into which purposeful errors were placed could be used as an instrument to evaluate peer reviewer performance. METHODS: An instrument for reviewer evaluation was created in the form of a fictitious manuscript into which deliberate errors were placed in order to develop an approach for the analysis of peer reviewer performance. The manuscript described a double-blind, placebo control study purportedly demonstrating that intravenous propranolol reduced the pain of acute migraine headache. There were 10 major and 13 minor errors placed in the manuscript. The work was distributed to all reviewers of Annals of Emergency Medicine for review. RESULTS: The manuscript was sent to 262 reviewers; 203 (78%) reviews were returned. One-hundred ninety-nine reviewers recommended a disposition for the manuscript: 15 recommended acceptance, 117 rejection, and 67 revision. The 15 who recommended acceptance identified 17.3% (95% confidence interval [CI] 11.3% to 23.4%) of the major and 11.8% (CI 7.3% to 16.3%) of the minor errors. The 117 who recommended rejection identified 39.1 % (CI 36.3% to 41.9%) of the major and 25.2% (CI 23.0% to 27.4%) of the minor errors. The 67 who recommended revision identified 29.6% (CI 26.1% to 33.1%) of the major and 22.0% (CI 19.3% to 24.8%) of the minor errors. The number of errors identified differed significantly across recommended disposition. Sixty-eight percent of the reviewers did not realize that the conclusions of the work were not supported by the results. CONCLUSION: These data suggest that the use of a preconceived manuscript into which purposeful errors are placed may be a viable approach to evaluate reviewer performance. Peer reviewers in this study failed to identify two thirds of the major errors in such a manuscript. PMID- 9737493 TI - Effect of attendance at a training session on peer reviewer quality and performance. AB - STUDY OBJECTIVE: To determine whether attendance at a voluntary training workshop improves quality ratings of medical journal peer reviewers. METHODS: Peer reviewers for Annals of Emergency Medicine who completed two or more reviews during the 20 months before or the 20 months after October 1995 were eligible. Reviews were routinely rated by editors on a subjective 5-point quality scale. Comparisons were made between reviewers who chose to attend a 4-hour workshop on peer review sponsored by the journal in 1995 (attendees) and 2 groups of reviewers who did not attend: controls matched for review quality and number of reviews completed before the workshop, and unmatched controls. Guest reviewers were excluded. RESULTS: A total of 298 reviewers completed 1906 reviews before the workshop and 2,194 after the workshop; 2,117 of these reviews were rated by editors. Forty-five attendees participated in the workshop, 39 of whom had sufficient ratings for analysis. Matched controls were almost identical in performance to attendees, but unmatched controls had performed fewer reviews and had lower average ratings before the workshop. There was no significant change in any performance measurement after the workshop, including average quality rating, percent change in quality rating, odds ratio for recommending acceptance, and odds ratio for congruence with editor's decision. CONCLUSION: In a self-selected group of experienced reviewers who attended a 4-hour workshop on peer review, no effect could be identified in subsequent performance as measured by editors' quality ratings or reviewer performance statistics. PMID- 9737494 TI - Absolute lymphocyte count as a predictor of CD4 count. AB - STUDY OBJECTIVE: To determine whether the absolute lymphocyte count (ALC) (white blood count x lymphocyte percentage) can be used to predict a low CD4 count. METHODS: We conducted a retrospective data analysis of consecutive CD4 count analyses performed between January 1, 1995, through December 1, 1995, at an urban university teaching hospital. Results of consecutive CD4 counts and simultaneously measured ALCs were analyzed from samples obtained in inpatient, clinic, and emergency department settings. The ability of ALC to predict a CD4 count less than 200 cells/mm3 was analyzed by calculating sensitivities, specificities, predictive values, and likelihood ratios for a range of ALC values. RESULTS: Among the 807 samples, 322 results (40%) had a CD4 count less than 200 cells/mm3. The ALC and CD4 count were correlated (r=.69, P<.0001). An ALC less than 1,000 cells/mm3 predicted CD4 counts less than 200 cells/mm3 with a sensitivity of .67 (95% confidence interval .62 to .72), specificity of .96 (.94 to .98), positive predictive value of .91 (.87 to .95), and a negative predictive value of .81 (.78 to .84). An ALC less than 2,000 cells/mm3 predicted CD4 counts less than 200 cells/mm3 with a sensitivity of .97 (.95 to .99), specificity of .41 (.37 to .45), positive predictive value of .52 (.48 to .56), and negative predictive value of .95 (.92 to .98). CONCLUSION: A reliable relationship exists between ALC and CD4 count. In a similar population, an ALC less than 1,000 cells/mm3 is predictive of a CD4 count less than 200 cells/mm3, and an ALC greater than or equal to 2,000 cells/mm3 is predictive of a CD4 count greater than or equal to 200 cells/mm3. Physicians may find these criteria useful in identifying patients with increased risk of opportunistic infection. PMID- 9737495 TI - Experimental tricyclic antidepressant toxicity: a randomized, controlled comparison of hypertonic saline solution, sodium bicarbonate, and hyperventilation. AB - STUDY OBJECTIVE: We sought to compare the effects of hypertonic sodium chloride solution (HTS), sodium bicarbonate solution, and hyperventilation (HV) on severe tricyclic antidepressant (TCA) toxicity in a swine model. METHODS: Twenty-four mixed-breed, domestic swine of either sex were given an intravenous infusion of nortriptyline (NT) until development of both a QRS duration longer than 120 ms and a systolic blood pressure (SBP) less than or equal to 50 mm Hg. Animals were randomly assigned to 1 of 4 groups. On reaching toxicity, the control group received 10 mL/kg of 5% dextrose in water (D5W); the HTS group received 10 mL/kg of 7.5% NaCl solution (15 mEq Na+/kg); the NaHCO3 group received 3 mEq/kg of 8.4% sodium bicarbonate solution followed by enough D5W solution to equal 10 mL/kg of total volume; and the HV group was mechanically hyperventilated to maintain arterial pH between 7.50 and 7.60 and given 10 mL/kg of D5W. RESULTS: The mean SBP 10 minutes after treatment was 54+/-18 mm Hg in the control group, 134+/-21 mm Hg in the HTS group, 85+/-19 mm Hg in the NaHCO3 group, and 60+/-12 mm Hg in the HV group (P<.05). Mean QRS duration 10 minutes after treatment was 144+/-38 ms in the control group, 80+/-14 ms in the HTS group, 105+/-38 ms in the NaHCO3 group, and 125+/-46 ms in the HV group (P<.05). CONCLUSION: In this model of TCA, toxicity HTS was more effective than sodium bicarbonate. Hyperventilation had little effect. Sodium loading may be the most important factor in reversing TCA toxicity. PMID- 9737496 TI - Randomized clinical trial of melatonin after night-shift work: efficacy and neuropsychologic effects. AB - OBJECTIVE: Melatonin has received considerable publicity for its sleep-promoting properties; however, there is little scientific evidence of its efficacy. The objective of this study is to determine whether there are measurable beneficial effects from exogenous melatonin in emergency physicians after intermittent night shift duty. METHODS: This randomized, placebo-controlled, double-blind, crossover trial was conducted in the emergency department of an urban tertiary care hospital. Fifteen emergency physicians were given melatonin 5 mg or placebo for 3 consecutive nights after night-shift duty with crossover to the opposite agent after a subsequent block of night shifts. The primary outcome measure was the global assessment of recovery measured by a visual analog scale. Secondary outcome measures included sleep quality, duration, and tiredness. In addition, the Profile of Mood States questionnaire and neuropsychologic testing were performed. RESULTS: There was no difference between melatonin and placebo in the global assessment of recovery (60.4+/-16.9 and 58.9+/-14.5, respectively; P=.29). There were no differences in sleep quality, duration or tiredness scores, sleep latency, hours of sleep obtained per night, and night or early awakening at any measurement point. Profile of Mood States and neuropsychologic test performances were similar. CONCLUSION: We found no beneficial effect of melatonin on sleep quality, tiredness, or cognitive function in emergency physicians after night shift duty. Our results suggest that exogenous melatonin is of limited value in recovery from night-shift work in emergency physicians. PMID- 9737497 TI - Indicators of assault-related injuries among women presenting to the emergency department. AB - STUDY OBJECTIVE: We sought to determine whether women presenting for treatment of assault-related injuries at a public hospital emergency department differed from those presenting for unintentional injuries with regard to a variety of demographic and presentation characteristics, nature and anatomic site of injury, and admission or follow-up treatment for injury. METHODS: We conducted a random sample retrospective medical record review of women aged 15 years and older who presented at either of 2 24-hour public-hospital emergency departments in Auckland, New Zealand. The characteristics of women identified as presenting with assault-related injuries on the basis of the record review were compared with those of women who presented for treatment of unintentional injuries. We also assessed the sensitivity and predictive value of nature and anatomic site of injury as markers of assault. RESULTS: We reviewed 8,051 records, of which 2,966 (37%) involved an injury at presentation. Two hundred sixty patients (9%) were identified as victims of assault. Of those women who presented with assault related injuries and had known assailants, most were likely injured by a partner or former partner. Women with assault-related injuries were more likely to be younger and of Maori or Pacific Islands origin. They were also more likely to present between the hours of 6 PM and 6 AM on Friday, Saturday, or Sunday and to have a greater history of prior presentations to the emergency department. Compared with patients who presented with unintentional injuries, women with assault-related injuries had a greater likelihood of presenting with contusions (odds ratio, 3.54; 95% confidence interval, 2.57 to 4.88); ill-defined signs and symptoms (odds ratio, 3.20; 95% confidence interval, 1.95 to 5.24); internal injuries (odds ratio, 2.48; 95% confidence interval, 1.46 to 4.18); fractures of the head, spine, or trunk (odds ratio, 2.09; 95% confidence interval, 1.23 to 3.53); and open wounds (odds ratio, 1.90; 95% confidence interval, 1.39 to 2.61). Assault-related injuries most commonly involved the head (odds ratio, 12.8; 95% confidence interval, 9.33 to 17.68). Despite the strength of these associations, however, with regard to nature of injury the sensitivity and positive predictive value of these indicators were limited (sensitivity < or = 26.5%, positive predictive value < or = 24.3%). The maximum sensitivity for anatomic site as a marker for assault was found for injuries to the head (63.7%), but the positive predictive value was still low at 35.7%. Women with assault-related injuries were more likely than women with unintentional injuries to be discharged from the emergency department without referral for follow-up treatment and were more likely to leave the department without completing treatment. CONCLUSION: Women identified as presenting with assault-related injuries differ from those who present with unintentional injuries in terms of their demographic and presentation characteristics, as well as the nature, anatomic site of injury, and follow-up treatment for injuries. Although some of this information has implications for service delivery to abused women, the use of clinical indicators such as nature and anatomic site of injury have limited predictive value. Therefore we recommend that health care providers routinely screen patients for assault, particularly assault by intimate partners, so that they may respond appropriately by providing better treatment and referral. PMID- 9737498 TI - Injured intoxicated drivers: citation, conviction, referral, and recidivism rates. AB - STUDY OBJECTIVES: Several studies have suggested that legally intoxicated drivers who are injured when involved in a motor vehicle crash are unlikely to be cited or prosecuted for driving under the influence (DUI). The purpose of this study was to determine (1) the rates of citation and prosecution of legally intoxicated drivers who are injured in a motor vehicle crash and hospitalized in a Level I trauma center, (2) the rates of previous and subsequent alcohol-related citation in this population, and (3) the rate of referral for treatment of alcohol-related problems made during the hospital stay. METHODS: In a retrospective review of trauma registry and Cleveland Municipal Court records from January 1993 through April 1995, we examined the records of all drivers injured in a motor vehicle crash who were transported to a Level I urban trauma center, admitted to the trauma service, and determined to have a blood alcohol content (BAC) of .10 gm% or higher at the time of admission to the emergency department. RESULTS: Seventy drivers admitted after a motor vehicle crash had a BAC of .10 gm% or higher. This represented 33% of the drivers older than 16 years of age who were admitted to the trauma service. Twenty-three drivers (32.8%) were cited for DUI, and 15 (21%) of the 70 were successfully prosecuted and convicted. Four of 23 cited drivers had previous citations; another 5 incurred subsequent citations during the study period. Eight of the 70 drivers who were admitted with a high BAC were referred for outpatient alcohol counseling after discharge. None were offered counseling as inpatients. CONCLUSION: Citation and prosecution rates of legally intoxicated drivers injured in motor vehicle crashes and hospitalized in our trauma center were low. Recognition of alcoholism and inpatient counseling were rare. Multiple alcohol-related citations were common among drivers cited for DUI. PMID- 9737500 TI - Marijuana and injury: is there a connection? PMID- 9737499 TI - Marijuana use and medically attended injury events. AB - STUDY OBJECTIVE: This study evaluated the relation between self-reported marijuana use and 3-year incidence of injury. METHODS: We conducted a retrospective cohort study of adult Kaiser Permanente Medical Care Program members who underwent multiphasic health examinations between 1979 and 1986 (n=4,462). Injury-related outpatient visits, hospitalizations, and fatalities within 3 years of examination were determined. RESULTS: Outpatient injury events totaled 2,524; 1,611 participants (36%) had at least 1 injury-related outpatient visit. Injury-related hospitalizations (n=22) and fatalities (n=3) were rare. Among men, there was no consistent relation between marijuana use and injury incidence for either former users (rate ratio, 1.15; 95% confidence interval [CI], .97 to 1.36) or current users (rate ratio, 0.97; 95% CI, .81 to 1.17), compared with those who had never used marijuana. Among women, former and current users showed little difference in their rate of later injury compared with never users; the rate ratios were 1.05 (95% CI, .87 to 1.26) and 1.20 (95% CI, 1.00 to 1.44), respectively. No statistically significant associations were noted between marijuana use and cause-specific injury incidence in men or women. CONCLUSION: Among members of a health maintenance organization, self-reported marijuana use in adult men or women was not associated with outpatient injury within 3 years of marijuana use ascertainment. PMID- 9737501 TI - Assault-related injury: what do we know, and what should we do about it? PMID- 9737502 TI - Emergency medicine in the new South Africa. AB - A description of recent developments in emergency medicine and prehospital care in the new South Africa is detailed. As South Africa creates a new integrated health care system with equal access for all citizens, expansion of emergency care services to previously disadvantaged populations is occurring. To illustrate current disparities, a comparison of 2 regional systems of emergency medicine is included. The challenges involved and possible future directions in this effort are discussed. PMID- 9737503 TI - Emergency care in Namibia. AB - Namibia is a sparsely populated nation in southwest Africa. A state-run health service provides care to most of the population. The geography and population distribution dictate the delivery systems for prehospital and emergency care. A state-run ambulance service provides basic patient transportation to the state run hospitals. There is no 911 system. Two private aeromedical companies in Namibia provide the full range of ground and aeromedical treatment, diver rescue, and helicopter and fixed-wing transport services. The scope of care includes cricothyrotomies, chest tubes, and rapid-sequence intubation. Equipment is modern and virtually identical to what is used in the United States. There are no emergency physicians in Namibia. General medical officers are the backbone of the state-run health service. General medical officers assigned to cover the ED are called casualty officers. No specialized training beyond internship is required, and assignments to casualty are viewed as temporary until better positions become available. Only the largest state hospital in the capital has a dedicated, 24 hour emergency staff. The private prehospital care/transport systems are well organized and sophisticated. Formal efforts should be undertaken to develop ties with our colleagues in Namibia. Potential areas for collaboration include injury surveillance and prevention, field trauma resuscitation, and prehospital care. PMID- 9737504 TI - Death by "ecstasy": the serotonin syndrome? AB - "Ecstasy" or 3,4-methylenedioxymethamphetamine (MDMA) is a popular drug of abuse and is generally regarded as safe by the lay public. There are an increasing number of reports of MDMA-induced toxicity that exhibit features of the serotonin syndrome. We report a case of severe hyperthermia, altered mental status, and autonomic dysfunction after a single recreational ingestion of MDMA. PMID- 9737505 TI - A novel source of carbon monoxide poisoning: explosives used in construction. AB - We describe an incident of carbon monoxide (CO) poisoning caused by CO migrating through soil after nearby detonation of explosive charges. Employees worked in a newly installed, unconnected manhole without incident and finished shortly before underground explosives were detonated 50 feet south of the manhole to break up rock and soil. A worker entering the manhole 45 minutes after the explosion collapsed within minutes, as did two coworkers who rescued him. One worker died, and all had elevated levels of carboxyhemoglobin. Air samples collected from the manhole 2 days after the incident showed 1,910 ppm CO; in laboratory detonations, sample explosive yielded 27 L CO per kilogram detonated. We believe the CO in this incident was released from the nearby explosion and migrated through soil and fractured rock into the manhole. The blasting and construction industries should be made aware of this previously unrecognized route of CO exposure. Additionally, confined-space procedures and training are needed to prevent future accidents. PMID- 9737506 TI - Tetanus among injecting-drug users--California, 1997. PMID- 9737507 TI - Thrombolysis for acute ischemic stroke. PMID- 9737508 TI - Radiologists' review of radiographs interpreted confidently by emergency physicians infrequently leads to change in patient management. PMID- 9737509 TI - Endotracheal intubation by basic EMTs. PMID- 9737510 TI - Bolus thrombolytic infusions during CPR. TPA in PEA Study Steering Committee. PMID- 9737511 TI - Atrial fibrillation and congestive heart failure: the intersection of two common diseases. PMID- 9737512 TI - Epidemiological and mechanistic studies of atrial fibrillation as a basis for treatment strategies. PMID- 9737513 TI - Impact of atrial fibrillation on the risk of death: the Framingham Heart Study. AB - BACKGROUND: Atrial fibrillation (AF) causes substantial morbidity. It is uncertain whether AF is associated with excess mortality independent of associated cardiac conditions and risk factors. METHODS AND RESULTS: We examined the mortality of subjects 55 to 94 years of age who developed AF during 40 years of follow-up of the original Framingham Heart Study cohort. Of the original 5209 subjects, 296 men and 325 women (mean ages, 74 and 76 years, respectively) developed AF and met eligibility criteria. By pooled logistic regression, after adjustment for age, hypertension, smoking, diabetes, left ventricular hypertrophy, myocardial infarction, congestive heart failure, valvular heart disease, and stroke or transient ischemic attack, AF was associated with an OR for death of 1.5 (95% CI, 1.2 to 1.8) in men and 1.9 (95% CI, 1.5 to 2.2) in women. The risk of mortality conferred by AF did not significantly vary by age. However, there was a significant AF-sex interaction: AF diminished the female advantage in survival. In secondary multivariate analyses, in subjects free of valvular heart disease and preexisting cardiovascular disease, AF remained significantly associated with excess mortality, with about a doubling of mortality in both sexes. CONCLUSIONS: In subjects from the original cohort of the Framingham Heart Study, AF was associated with a 1.5- to 1.9-fold mortality risk after adjustment for the preexisting cardiovascular conditions with which AF was related. The decreased survival seen with AF was present in men and women and across a wide range of ages. PMID- 9737514 TI - Assessment of atrioventricular junction ablation and VVIR pacemaker versus pharmacological treatment in patients with heart failure and chronic atrial fibrillation: a randomized, controlled study. AB - BACKGROUND: Uncontrolled studies have suggested that atrioventricular junction ablation and pacemaker implantation have beneficial effects on quality of life in patients with chronic atrial fibrillation (AF). METHODS AND RESULTS: We performed a multicenter, controlled, randomized, 12-month evaluation of the clinical effects of atrioventricular junction ablation and VVIR pacemaker (Abl+Pm) versus pharmacological (drug) treatment in 66 patients with chronic (lasting >6 months) AF who had clinically manifest heart failure and heart rate >90 bpm on 3 standard ECGs recorded at rest during stable clinical conditions on different days. Before completion of the study, withdrawals occurred in 8 patients of the drug group and in 4 patients of the Abl+Pm group. At the end of the 12 months, the 28 Abl+Pm patients who completed the study showed lower scores in palpitations (-78%; P=0.000) and effort dyspnea (-22%; P=0.05) than the 26 of the drug group. Lower scores, although not significant, were also observed for exercise intolerance ( 20%), easy fatigue (-17%), chest discomfort (-50%), Living with Heart Failure Questionnaire (-14%), New York Heart Association functional classification (-4%), and Activity scale (-12%). The intrapatient comparison between enrollment and month 12 showed that in the Abl+Pm group, all variables except easy fatigue improved significantly from 14% to 82%. However, because an improvement was also observed in the drug group, the difference between the 2 groups was significant only for palpitations (P=0.000), effort dyspnea (P=0.01), exercise intolerance (P=0.005), easy fatigue (P=0.02), and chest discomfort (P=0.02). Cardiac performance, evaluated by means of standard echocardiogram and exercise test, did not differ significantly between the 2 groups and remained stable over time. CONCLUSIONS: In patients with heart failure and chronic AF, Abl+Pm treatment is effective and superior to drug therapy in controlling symptoms, although its efficacy appears to be less than that observed in uncontrolled studies because some improvement can also be expected in medically treated patients. Cardiac performance is not modified by the treatment. PMID- 9737515 TI - Cardiac sympathetic dysinnervation in diabetes: implications for enhanced cardiovascular risk. AB - BACKGROUND: Regional cardiac sympathetic hyperactivity predisposes to malignant arrhythmias in nondiabetic cardiac disease. Conversely, however, cardiac sympathetic denervation predicts increased morbidity and mortality in severe diabetic autonomic neuropathy (DAN). To unite these divergent observations, we propose that in diabetes regional cardiac denervation may elsewhere induce regional sympathetic hyperactivity, which may in turn act as a focus for chemical and electrical instability. Therefore, the aim of this study was to explore regional changes in sympathetic neuronal density and tone in diabetic patients with and without DAN. METHODS AND RESULTS: PET using the sympathetic neurotransmitter analogue 11C-labeled hydroxyephedrine ([11C]-HED) was used to characterize left ventricular sympathetic innervation in diabetic patients by assessing regional disturbances in myocardial tracer retention and washout. The subject groups comprised 10 diabetic subjects without DAN, 10 diabetic subjects with mild DAN, 9 diabetic subjects with severe DAN, and 10 healthy subjects. Abnormalities of cardiac [11C]-HED retention were detected in 40% of DAN-free diabetic subjects. In subjects with mild neuropathy, tracer defects were observed only in the distal inferior wall of the left ventricle, whereas with more severe neuropathy, defects extended to involve the distal and proximal anterolateral and inferior walls. Absolute [11C]-HED retention was found to be increased by 33% (P<0.01) in the proximal segments of the severe DAN subjects compared with the same regions in the DAN-free subjects (30%; P<0.01 greater than the proximal segments of the mild DAN subjects). Despite the increased tracer retention, no appreciable washout of tracer was observed in the proximal segments, consistent with normal regional tone but increased sympathetic innervation. Distally, [11C] HED retention was decreased in severe DAN by 33% (P<0.01) compared with the DAN free diabetic subjects (21%; P<0.05 lower than the distal segments of the mild DAN subjects). CONCLUSIONS: Diabetes may result in left ventricular sympathetic dysinnervation with proximal hyperinnervation complicating distal denervation. This combination could result in potentially life-threatening myocardial electrical instability and explain the enhanced cardioprotection from beta blockade in these subjects. PMID- 9737516 TI - Increased availability and open probability of single L-type calcium channels from failing compared with nonfailing human ventricle. AB - BACKGROUND: The role of the L-type calcium channel in human heart failure is unclear, on the basis of previous whole-cell recordings. METHODS AND RESULTS: We investigated the properties of L-type calcium channels in left ventricular myocytes isolated from nonfailing donor hearts (n= 16 cells) or failing hearts of transplant recipients with dilated (n=9) or ischemic (n=7) cardiomyopathy. The single-channel recording technique was used (70 mmol/L Ba2+). Peak average currents were significantly enhanced in heart failure (38.2+/-9.3 fA) versus nonfailing control hearts (13.2+/-4.5 fA, P=0.02) because of an elevation of channel availability (55.9+/-6.7% versus 26.4+/-5.3%, P=0.001) and open probability within active sweeps (7.36+/-1.51% versus 3.18+/-1.33%, P=0.04). These differences closely resembled the effects of a cAMP-dependent stimulation with 8-Br-cAMP (n= 11). Kinetic analysis of the slow gating shows that channels from failing hearts remain available for a longer time, suggesting a defect in the dephosphorylation. Indeed, the phosphatase inhibitor okadaic acid was unable to stimulate channel activity in myocytes from failing hearts (n=5). Expression of calcium channel subunits was measured by Northern blot analysis. Expression of alpha1c- and beta-subunits was unaltered. Whole-cell current measurements did not reveal an increase of current density in heart failure. CONCLUSIONS: Individual L type calcium channels are fundamentally affected in severe human heart failure. This is probably important for the impairment of cardiac excitation-contraction coupling. PMID- 9737517 TI - Complex demodulation of cardiorespiratory dynamics preceding vasovagal syncope. AB - BACKGROUND: The dynamic autonomic processes leading to vasovagal syncope are poorly understood. METHODS AND RESULTS: We used complex demodulation to continuously assess changes in respiration, R-R interval, and arterial pressure (blood pressure) variability during 60 degree head-up tilt in 25 healthy subjects with tilt-induced vasovagal syncope and 25 age-matched nonsyncopal control subjects. Coherence and transfer function analyses were used to examine the relation between respiration and R-R interval variability before syncope. Baseline blood pressure, R-R, and ventilation were similar between syncope subjects and control subjects. Syncope subjects experienced an increase in tidal volume and decrease in BP beginning 3 minutes before impending syncope (systolic blood pressure <80 mm Hg) necessitated termination of tilt. Approximately 90 seconds before syncope there was a sudden prolongation of R-R interval and increase in amplitude of high and low frequency R-R interval variability, indicating an abrupt enhancement of vagal tone. The increase in respiratory amplitude between 180 and 90 seconds before syncope was not accompanied by changes in R-R interval or R-R variability, suggesting a dissociation between respiration and the respiratory sinus arrhythmia. The coherence analysis showed fewer syncope subjects with coherence between respiratory and R-R interval variabilities and lower transfer magnitudes in syncope subjects compared with control subjects. Nonsyncopal subjects had no change in respiratory, R-R interval, or blood pressure dynamics during matched time periods before the time of syncope. CONCLUSIONS: Vasovagal syncope is preceded by a period of hyperpnea and cardiorespiratory decoupling followed by an abrupt increase in cardiovagal tone. Respiratory pumping without inspiratory cardiac slowing may partially counteract preload reduction until sudden bradycardia precipitates syncope. PMID- 9737518 TI - Spiral computed tomography: a novel diagnostic approach for investigation of the extracranial cerebral arteries and its complementary role in duplex ultrasonography. AB - BACKGROUND: For the detection of atherosclerotic lesions of the extracranial cerebral arteries, duplex ultrasonography (US) is an established operator dependent method, whereas arteriography is associated with the not-insignificant risk of embolic complications. Spiral CT is a promising novel diagnostic tool that allows noninvasive, operator-independent diagnosis of obstruction of extracranial cerebral arteries. The aim of our prospective study was to evaluate in a clinical setting the complementary role of duplex US and spiral CT. METHODS AND RESULTS: We compared the results obtained independently by spiral CT and duplex US in 59 consecutive patients with clinical suspicion of an obstructive lesion affecting the carotid arteries. We analyzed a total of 354 segments from the extracranial carotid arteries, including the common, internal, and external carotid arteries. A total of 4 complete occlusions, 38 severe stenoses (70% to 99%), and 32 moderate stenoses (30% to 69%) were concordantly identified by means of duplex US and spiral CT. In 5 cases in which duplex US did not allow sufficient evaluation of the carotid artery because of a poor US window or severe calcification, spiral CT allowed identification and correct measurement of the stenotic lesion. The comparison of the percentage of stenosis with both methods was good (r=0.91, P=0.024). CONCLUSIONS: Our results indicate that spiral CT of the extracranial cerebral arteries is a promising noninvasive complementary and non-operator-dependent examination. Its application is particularly attractive in cases in which duplex US is not reliable (ie, severe kinking, severe calcification, short neck, and high bifurcation) and particularly when an overall view of the vascular field is required. PMID- 9737519 TI - Prognostic value of dipyridamole-thallium myocardial scintigraphy in patients with Kawasaki disease. AB - BACKGROUND: Although coronary artery lesions are critical complications of Kawasaki disease, their long-term outcome is still unclear. It is sometimes difficult to monitor progressive changes from aneurysms to stenotic lesions because coronary angiography (CAG) cannot be repeated very often, especially in infants. Our prospective study was designed to evaluate the prognostic value of dipyridamole-thallium single-photon-emission CT (SPECT) in the long-term follow up of patients with Kawasaki disease. METHODS AND RESULTS: Of 459 consecutive patients with Kawasaki disease, coronary aneurysms were detected in 90 cases by echocardiography during the acute stage. After paired studies of selective CAG and SPECT were conducted, all patients were followed up and monitored for the occurrence of any cardiac events for > or =8 years. During the follow-up interval, there were 15 cardiac events (1 death, 5 infarctions, 2 coronary artery bypass graft operations, and 7 occurrences of unstable angina). Of patients who had some event, thallium redistribution was found on SPECT in 14 (93%, P<0.001). Of the various clinical and scintigraphic image variables, the presence of thallium redistribution was the best multivariate independent predictor of a late cardiac event (chi2=57.8, P<0.0001). The number of aneurysms detected on CAG added minimal statistical improvement to the model (chi2=1.9, P=0.0009). CONCLUSIONS: Dipyridamole-thallium SPECT is safely performed and is useful and important for risk stratification in the long-term follow-up of patients with Kawasaki disease. PMID- 9737520 TI - Progressive tricuspid valve disease in patients with congenitally corrected transposition of the great arteries. AB - BACKGROUND: The outcome of patients with corrected transposition of the great arteries (CTGA) is variably affected by associated intracardiac defects, tricuspid valve competence, and systemic right ventricular (RV) function. The relative importance of these factors in long-term outcome has not been evaluated. METHODS AND RESULTS: Since 1958, 40 patients with CTGA were studied to determine risk factors for poor outcome, including age, open heart surgery, tricuspid insufficiency (TI), cardiac rhythm, pulmonary overcirculation, and RV dysfunction. Follow-up ranged from 7 to 36 years (mean 20 years). Intracardiac repair was performed in 21 patients; 19 were unoperated or had closed heart procedures. For the purposes of this study the designation TI(S) refers to at least moderately severe TI as delineated by echocardiogram and/or angiography. TI(s) was the only independently significant factor for death (P=0.01), and in turn, only the presence of a morphologically abnormal TV predicted TI(s)(P=0.03). Twenty-year survival without TI(S)was 93%, but only 49% with TI(S). Poor long term postoperative outcome was due to TI(S) in all but 1 patient; 20-year survival rates for operated patients with and without TI(s)were 34% and 90%, respectively (P=0.002). Similarly, 20-year survival rates for unoperated patients with and without TI(s)were 60% and 100%, respectively, whether or not attempts to repair the TI were made (P=0.08). CONCLUSIONS: TI(S)represents the major risk factor for CTGA patients; RV dysfunction appears to be almost always secondary to long-standing TI. Decisions related to surgical interventions with or without associated lesions must be strongly influenced by the status of the tricuspid valve. PMID- 9737521 TI - Magnitude and time course of microvascular obstruction and tissue injury after acute myocardial infarction. AB - BACKGROUND: Microvascular obstruction within an area of myocardial infarction indicates worse functional recovery and a higher risk of postinfarction complications. After prolonged coronary occlusion, contrast-enhanced MRI identifies myocardial infarction as a hyperenhanced region containing a hypoenhanced core. Because the time course of microvascular obstruction after infarction/reperfusion is unknown, we examined whether microvascular obstruction reaches its full extent shortly after reperfusion or shows significant progression over the following 2 days. METHODS AND RESULTS: Seven dogs underwent 90-minute balloon occlusion of the left anterior descending coronary artery (LAD) followed by reflow. Gadolinium-DTPA-enhanced MRI performed at 2, 6, and 48 hours after reperfusion was compared with radioactive microsphere blood flow (MBF) measurements and myocardial staining to define microvascular obstruction (thioflavin S) and infarct size (triphenyltetrazolium chloride, TTC). The MRI hypoenhanced region increased 3-fold during 48 hours after reperfusion (3.2+/ 1.8%, 6.7+/-4.4%, and 9.9+/-3.2% of left ventricular mass at 2, 6, and 48 hours, respectively, P<0.03) and correlated well with microvascular obstruction (MBF <50% of remote region, r=0.99 and thioflavin S, r=0.93). MRI hyperenhancement also increased (21.7+/-4.0%, 24.3+/-4.6%, and 28.8+/-5.1% at 2, 6, and 48 hours, P<0.006) and correlated well with infarct size by TTC (r=0.92). The microvascular obstruction/infarct size ratio increased from 13.0+/-4.8% to 22.6+/-8.9% and to 30.4+/-4.2% over 48 hours (P=0.024). CONCLUSION: The extent of microvascular obstruction and the infarct size increase significantly over the first 48 hours after myocardial infarction. These results are consistent with progressive microvascular and myocardial injury well beyond coronary occlusion and reflow. PMID- 9737522 TI - Determination of right ventricular structure and function in normoxic and hypoxic mice: a transesophageal echocardiographic study. AB - BACKGROUND: Noninvasive cardiac evaluation is of great importance in transgenic mice. Transthoracic echocardiography can visualize the left ventricle well but has not been as successful for the right ventricle (RV). We developed a method of transesophageal echocardiography (TEE) to evaluate murine RV size and function. METHODS AND RESULTS: Normoxic and chronically hypoxic mice (F(IO2)=0.11, 3 weeks) and agarose RV casts were scanned with a rotating 3.5F/30-MHz intravascular ultrasound probe. In vivo, the probe was inserted in the mouse esophagus and withdrawn to obtain contiguous horizontal planes at 1-mm intervals. In vitro, the probe was withdrawn along the left ventricular posterior wall of excised hearts. The borders of the RV were traced on each plane, allowing calculation of diastolic and systolic volumes, RV mass, RV ejection fraction, stroke volume, and cardiac output. RV wall thickness was measured. Echo volumes obtained in vitro were compared with cast volumes. Echo-derived cardiac output was compared with measurements of an ascending aortic Doppler flow probe. Echo-derived RV free wall mass was compared with true RV free wall weight. There was excellent agreement between cast and TEE volumes (y=0.82x+6.03, r=0.88, P<0.01) and flow-probe and echo cardiac output (y=1.00x+0.45, r=0.99, P<0.0001). Although echo-derived RV mass and wall thickness were well correlated with true RV weight, echo-derived RV mass underestimated true weight (y=0.53x+2.29, r=0.81, P<0.0001). RV mass and wall thickness were greater in hypoxic mice than in normoxic mice (0.78+/-0.19 versus 0.51+/-0.14 mg/g, P<0.03, 0.50+/-0.03 versus 0.38+/-0.03 mm, P<0.04). CONCLUSIONS: TEE with an intravascular ultrasound catheter is a simple, accurate, and reproducible method to study RV size and function in mice. PMID- 9737523 TI - Ischemic preconditioning in pigs: a graded phenomenon: its relation to adenosine and bradykinin. AB - BACKGROUND: A threshold concept for ischemic preconditioning (IPc) has been proposed. It is unclear, however, whether IPc, above a certain threshold, is an all-or-nothing or a graded phenomenon. METHODS AND RESULTS: In 71 enflurane anesthetized swine, severe left anterior descending coronary artery hypoperfusion for 90 minutes followed by 2 hours of reperfusion resulted in an infarct size (IS, by triphenyltetrazolium chloride) of 16.7+/-3.4% (SEM) of the area at risk. IPc by 2 minutes of low-flow ischemia and 15 minutes of reperfusion before the 90 minute target ischemia did not reduce IS (21.9+/-7.0%). IS was decreased to 9.0+/ 2.6% (P<0.05) by 3 minutes of IPc and reduced further to 1.9+/-0.9% (P<0.05) by 10 minutes of IPc. The interstitial adenosine concentration (microdialysis, high performance liquid chromatography) was unchanged with 2 and 3 minutes of IPc but increased with 10 minutes of IPc (by 573+/-144%). The interstitial bradykinin concentration (microdialysis, radioimmunoassay) remained unchanged with 2 minutes of IPc but increased to a similar extent with 3 minutes (by 198+/-32%) and 10 minutes (by 224+/-30%) of IPc. The IS reduction by 3 minutes of IPc was abolished by blockade of the bradykinin B2 receptor with intracoronary HOE 140 (16.6+/ 4.3%) but not with intracoronary infusion of adenosine deaminase (8.4+/-2.5%, P<0.05). HOE 140, however, did not affect the IS reduction (3.5+/- 1.1%, P<0.05) by 10 minutes of IPc. Combined infusion of HOE 140 and adenosine deaminase abolished the IS reduction by 10 minutes of IPc (15.4+/-6.7%). CONCLUSIONS: IS reduction by IPc is a graded phenomenon. Whereas bradykinin is essential during preconditioning ischemia of shorter duration, adenosine is more important during preconditioning ischemia of longer duration. PMID- 9737524 TI - Enhancement of fibrinolysis with 40-kHz ultrasound. AB - BACKGROUND: Ultrasound at frequencies of 0.5 to 1 MHz and intensities of > or =0.5 W/cm2 accelerates enzymatic fibrinolysis in vitro and in some animal models, but unacceptable tissue heating can occur, and limited penetration would restrict application to superficial vessels. Tissue heating is less and penetration better at lower frequencies, but little information is available regarding the effect of lower-frequency ultrasound on enzymatic fibrinolysis. We therefore examined the effect of 40-kHz ultrasound on fibrinolysis, tissue penetration, and heating. METHODS AND RESULTS: 125I-fibrin-radiolabeled plasma clots in thin-walled tubes were overlaid with plasma containing tissue plasminogen activator (tPA) and exposed to ultrasound. Enzymatic fibrinolysis was measured as solubilization of radiolabel. Tissue attenuation and heating were examined in samples of porcine rib cage. Fibrinolysis was increased significantly in the presence of 40-kHz ultrasound at 0.25 W/cm2, reaching 39+/-7% and 93+/-11% at 60 minutes and 120 minutes, compared with 13+/-8% and 37+/-4% in the absence of ultrasound (P<0.0001). The acceleration of fibrinolysis increased at higher intensities. Attenuation of the ultrasound field was only 1.7+/-0.5 dB/cm through the intercostal space and 3.4+/-0.9 dB/cm through rib. Temperature increments in rib were <1 C/(W/cm2). CONCLUSIONS: These findings indicate that 40-kHz ultrasound significantly accelerates enzymatic fibrinolysis at intensities of > or =0.25 W/cm2 with excellent tissue penetration and minimal heating. Externally applied 40-kHz ultrasound at low intensities is a potentially useful therapeutic adjunct to enzymatic fibrinolysis with sufficient tissue penetration for both peripheral vascular and coronary applications. PMID- 9737525 TI - Images in cardiovascular medicine. Unusual left ventricular mass. PMID- 9737526 TI - Images in cardiovascular medicine. Left main coronary artery disease: cardiac arrest following stress echocardiography. PMID- 9737527 TI - Attenuation of unfavorable sympathetic hyperactivity induced by long-term physical training in postinfarction patients: fact or speculation? PMID- 9737528 TI - Lung function and exercise gas exchange in chronic heart failure. PMID- 9737529 TI - Hepatitis C virus infection and chronic active myocarditis. PMID- 9737530 TI - Prediction of transition to chronic atrial fibrillation in patients with paroxysmal atrial fibrillation. PMID- 9737531 TI - Conservation of eukaryotic DNA repair mechanisms. AB - PURPOSE: To discuss the evolutionary conservation of different DNA repair processes. The proteins that carry out base excision repair show a varying degree of structural conservation, but a high level of functional complementation between species, as might be expected for a sequential pathway. In nucleotide excision repair there is a high degree of structural conservation, but few examples of functional complementation because the process involves multiprotein complexes. Repair by homologous recombination involves proteins that are highly conserved structurally. The process of repair of DNA breaks by non-homologous end joining is conserved in eukaryotes, but the level of sequence identity of several of the proteins is fairly low and some components involved in man do not appear to have sequence homologues in yeast. CONCLUSIONS: All DNA repair processes are highly conserved. The degree of structural and functional conservation varies between the different processes. PMID- 9737533 TI - Frequencies and types of exchange aberrations induced by X-rays and neutrons in Chinese hamster splenocytes detected by FISH using chromosome-specific DNA libraries. AB - PURPOSE: To estimate the frequencies of radiation- (low and high LET) induced chromosome aberrations in Chinese hamster splenocytes by two-colour fluorescence in situ hybridization using DNA painting probes specific for chromosomes 2, 3, 8, X and Y and to determine (1) the ratio of radiation-induced translocations and dicentrics; (2) the spectrum of exchange aberrations induced by X-rays and neutrons; and (3) the relative involvement of the different chromosomes in the formation of aberrations. MATERIALS AND METHODS: Isolated splenocytes from the Chinese hamster were irradiated in vitro with different doses of 200 kV X-rays (0.75, 1.5, 3.0 Gy) and 1 MeV fast neutrons (0.25, 0.5, 1.0 Gy). Conventional analysis of chromosome aberrations was carried out in Giemsa-stained preparations. Chromosome aberrations involving chromosomes 2, 3, 8, X and Y were analysed in first division metaphases using two-colour FISH. RESULTS: The results indicate that when all types of translocations are taken into account both X-rays and neutrons induce more translocations than dicentrics, the ratio between the two types of exchanges being 1.4 and 1.8 respectively. The ratio of 'apparently simple' reciprocal translocations and reciprocal complete dicentrics was close to 1 for both types of radiation. The RBE of neutrons for induction of exchanges was found to be between 5 and 8. Neutron irradiation was more efficient at inducing insertions. Among the chromosomes studied, an increased involvement was observed for chromosome 8 in dicentrics and translocations than that expected on the basis of its chromosome length. The high content of interstitial telomeric sequences in chromosome 8 may be responsible for the observed sensitivity of this chromosome. CONCLUSIONS: The results obtained in this study indicate that: (1) more translocations are found than dicentrics; (2) heterogeneity exists among Chinese hamster chromosomes for involvement in radiation-induced exchanges; (3) the spectrum and distribution of exchange aberrations are different between X-rays and neutrons; and (4) the relative frequencies of insertions could be used as a 'fingerprint' for exposure to high LET radiation. PMID- 9737532 TI - The p53-mediated DNA damage response to ionizing radiation in fibroblasts from ataxia-without-telangiectasia patients. AB - PURPOSE: To assess the functionality of the p53-mediated pathway, activated by the ataxia-telangiectasia gene product (ATM) in response to ionizing radiation, in cells derived from four ataxia-without-telangiectasia patients. These patients exhibit cerebellar ataxia and cellular abnormalities that are compatible with the diagnosis of ataxia-telangiectasia (AT), but the telangiectasias normally seen in AT patients are absent. MATERIALS AND METHOD: Protein and RNA extracts were prepared from primary fibroblast cultures non- or exposed to 5 Gy of ionizing radiation in order to monitor the modulation in p53 and ATM protein levels by immunologic techniques and WAF1/Cip1(p21) mRNA by Northern blotting. RESULTS: A sub-optimal response in terms of increased levels of p53 and the transcriptional activation of WAF1/Cip1(p21) was see in the ataxia-without-telangiectasia fibroblast cultures examined over a 4 h period post-irradiation when compared with normal fibroblast cultures. The ATM protein was expressed at much reduced levels in the ataxia-without-telangiectasia and the classical AT fibroblast cultures examined when compared with normal fibroblast cultures. CONCLUSIONS: Despite the milder clinical phenotypes observed in these ataxia-without telangiectasia patients and the presence of low levels of ATM protein in the fibroblast cultures, their response to ionizing radiation quantitatively resembles that reported in fibroblast cultures established from classical AT patients. PMID- 9737534 TI - DNA-proportional distribution of radiation-induced chromosome aberrations analysed by fluorescence in situ hybridization painting of all chromosomes of a human female karyotype. AB - PURPOSE: This is the extension of a previous study, showing deviations from a DNA proportional involvement of 12 single chromosomes (1-4, 6-10, 12, 14 and X) in radiation-induced translocations and dicentrics measured by FISH-painting and classified by standard cytogenetic scoring criteria. By adding data on chromosomes 2, 4, 5, 9, 11-13, 15-22 and X the analysis now comprises all chromosomes of a human female karyotype evaluated with three nomenclature systems (PAINT, S & S and a conventional method). MATERIAL AND METHODS: Metaphase spreads were prepared from lymphocytes irradiated with 3 Gy 220 kV X-rays. FISH painting was performed with single chromosome-specific probes in combination with a pancentromeric probe. RESULTS: Deviations from a DNA-proportional distribution became apparent for all aberration parameters analysed with the three nomenclature systems. Chromosomes 2, 3 and 6 were less frequently involved and chromosomes 16, 17 and 20 were more frequently involved in exchange aberrations. Generally, smaller chromosomes (15-22, with the exception of chromosome 19) were more frequently involved in aberration formation than expected. CONCLUSION: The assumption that the probability of a chromosome being involved in an exchange aberration is proportional to its DNA content is not supported by the present data. PMID- 9737535 TI - Analysis of chromosome damage based on the time course of aberrations. AB - PURPOSE: To describe a method for the analysis of radiation induced chromosome aberrations which will in particular be suitable for the comparison of different radiation qualities. The method is applied to previously published data. METHODS: The basic idea of the approach is to use the time-course of aberrations up to the time when all cells have passed through first mitosis. Instead of comparing data obtained at a single sampling time, the number of aberrant cells or aberrations is integrated over several sampling times. The integral represents a measure of the total fraction of cells showing aberrations and the total amount of aberrations induced in the irradiated cell population. In addition, a correction factor is applied which takes into account the dilution of heavily damaged cells at late sampling times by cell division of undamaged or less severely damaged cells. RESULTS AND CONCLUSIONS: Previously published data on the induction of chromosome aberrations by 4.6 MeV/u Ar ions and X-rays are re-analysed using the new approach. It is demonstrated that this method allows a better comparison of experiments using extremely different radiation qualities and consequently different cell cycle perturbations. PMID- 9737536 TI - A pulse radiolysis study of carnosine in aqueous solution. AB - PURPOSE: To investigate the potential antioxidative role of carnosine (beta alanyl-L-histidine) through its interaction with radiation induced radicals. MATERIALS AND METHODS: Pulse radiolysis experiments were performed by a 12 MeV electron linear accelerator (LINAC) on carnosine aqueous solutions at different pHs. The Raman spectra of solid samples were obtained by a Bruker IFS 66 spectrometer. RESULTS: The protonation state of the imidazole ring was observed to affect the site(s) of .OH attack on carnosine as well as the decay rates of the resulting adducts. Reasonably, a base catalysed water elimination from the adducts leading to a resonance-stabilized radical, which absorbs at lambda = 270 nm, takes place. Raman spectra in slightly alkaline medium have indicated that carnosine is mainly present as tautomer I and, of consequence, positions C(2) and C(4) of the imidazole ring are the preferential sites for .OH attack. CONCLUSIONS: These studies have shown that carnosine is a good scavenger of .OH radicals giving rise to a quite stable intermediate which should be less reactive towards other biological components than .OH itself. Raman data have been helpful in predicting the preferential sites for the .OH attack. PMID- 9737537 TI - Free radicals from X-irradiated 'dry' and hydrated lyophilized DNA as studied by electron spin resonance spectroscopy: analysis of spectral components between 77K and room temperature. AB - PURPOSE: To investigate the number, spectroscopic signatures and chemical structures of free radicals from X-irradiated lyophilized DNA (dry and equilibrated at 76% relative humidity) between 77 K and room temperature by electron spin resonance (ESR) spectroscopy. MATERIALS AND METHODS: Samples were prepared by freeze drying DNA (sodium salt, salmon testes) in H2O or D2O and used as such ('dry' DNA) or after equilibration at 76% relative humidity. K3[Fe(CN)6] was co-lyophilized in some samples as an electron scavenger. X-irradiation was performed at 77 K (liquid nitrogen). Data acquisition was on a Bruker ESP 380 ESR spectrometer (X-band, 9.5 GHz) and at high magnetic fields (245 GHz, Y-band; GHMFI, Grenoble, France). Data analysis involved computer treatment of spectra. RESULTS: There were 12 different radical components isolated from DNA in four different conditions (dry and after equilibration at 76% relative humidity in either H2O or D2O) with the additional help of high magnetic field ESR and the use of K3[Fe(CN)6] as an electron scavenger. Several components were detected at 77 K and were found to be common for both hydration conditions, although their spectral shape varied considerably. These involved reduced thymine and cytosine bases, the oxidized guanine base, probably a C1'-located sugar radical, a thymine allyl radical and a secondary thymine H-addition radical. For the reduced cytosine base the amino-protonated form was observed in H2O samples, which was only partially exchanged in the D2O samples. At high water content another species, perhaps due to a sugar radical, contributes in addition even at low temperatures. All radical components anneal out with temperature, with only small secondary reactions taking place. A peroxy radical and a sharp singlet, probably due to the deprotonated radical cation from guanine, come into the balance together with the secondary thymine radical. At high doses, a further sugar radical (perhaps at the C3'-position) was detected in dry DNA. The relative yields of the isolated patterns were determined by precise reconstruction of the experimental spectra. CONCLUSIONS: The comprehensive component delineation performed at 77 K and upon annealing to room temperature for lyophilized DNA showed a larger diversity and a higher variance of radicals at 77 K than discussed so far. Thermal annealing brings about only a few reactions to produce secondary species. Most components decay without paramagnetic successors. PMID- 9737538 TI - Thiyl radical-induced cis/trans-isomerization of methyl linoleate in methanol and of linoleic acid residues in liposomes. AB - PURPOSE: To investigate the role of a thiol-containing biologically active compound in lipid peroxidation of membranes. MATERIALS AND METHODS: Thiyl radicals were generated from 3-(2-mercaptoethyl)quinazoline-2,4(1H,3H)-dione (MECH) using pulse radiolysis and gamma-radiolysis in aqueous and alcoholic solutions saturated with N2O. The products were analysed by 1H NMR and by HPLC. RESULTS: THE thiyl radicals abstract bisallylic hydrogens from [cis-9, cis-12] methyl linoleate, yielding a pentadienyl radical. In the absence of oxygen, a thiyl radical-induced cis/trans-isomerization leads to linoleic-type isomers. These chain-type isomerization reactions can occur with the long living pentadienyl radical, followed by a 'repair' reaction of the attached thiol, and with the thiyl radical adduct with a double bond of the fatty acid residue. CONCLUSIONS: The results show that the mechanism of cis/trans-isomerization is an integral part of the thiyl radical attack on polyunsaturated fatty acids in homogeneous solutions and in bilayers. PMID- 9737540 TI - Characterization of the specific response to serotonin of mouse tumour-feeding arterioles. AB - PURPOSE: To investigate the role of tumour versus non-tumour factors in the specific response to serotonin (5-HT) of tumour-feeding arterioles (TFA). MATERIALS AND METHODS: Using mouse models of intra-vital microscopy, the response to topical administration of 5-HT was studied in arterioles feeding tumours: fibrosarcoma (Meth A), murine mammary adenocarcinoma (EMT6) and human colo-rectal carcinoma (HRT18) intra-cutaneously implanted. RESULTS: For all types of tumour, 5-HT induced a far more pronounced constriction of TFA than of control arterioles. The presence of a tumour implanted in the connective tissue between the skin and the cremaster muscle also affected the reactivity of muscle arterioles. Conversely, the response to serotonin by neovessels grown after implantation of an exogenous element under the skin did not differ from that of control arterioles. CONCLUSIONS: Changes in reactivity to serotonin were not dependent on the type of tumour implanted in the skin and were not present for a non-tumour implant. The presence of the tumour can alter the reactivity of vessels from tissue in contact with the tumour even if these vessels did not feed the tumour. This phenomenon is local and was not found in the vessels at a distance from the tumour. PMID- 9737539 TI - Mechanisms of the anti-inflammatory activity of low-dose radiation therapy. AB - PURPOSE: To investigate the hypothesis that modulation of the function of activated macrophages is one of the mechanisms of the clinically observed anti inflammatory and analgesic efficacy of low-dose radiotherapy in the treatment of a variety of painful joint diseases with total doses between 1 and 6 Gy. MATERIALS AND METHODS: Metabolic activity, cell proliferation, reproductive integrity, nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) expression by unstimulated or [LPS/gamma-IFN-stimulated macrophages in vitro was investigated at different times after radiation doses ranging from 0.3 Gy to 10 Gy. In vivo, chronic granulomatous air pouches were induced in mice and either sham treated or irradiated with 2 Gy on day 2 or day 6, or with five daily doses of 0.5 Gy. On day 7, the iNOS expression was assessed by Western blot and localized by immuno-histochemistry in cryostat sections. RESULTS: In stimulated macrophages, metabolic activity, proliferation and reproductive integrity were not affected by radiation doses up to 10 Gy since they are apparently irreversible post-mitotic cells. However, a dose-dependent modulation of the NO pathway was observed with significant inhibition by the low radiation doses used in anti-inflammatory radiotherapy but with super-stimulation by the high radiation doses used in cancer therapy. CONCLUSIONS: The empirically based anti inflammatory radiotherapy of benign diseases appears to act through specific modulation of different pathways of inflammatory reactions such as the nitric oxide pathway in stimulated macrophages. PMID- 9737541 TI - Human thymic stromal cell irradiation reduces intra-thymic T cell precursor proliferation: evidence for a soluble mediator. AB - PURPOSE: To investigate the effects of ionizing radiations on human thymic stromal cells and the consequences of stromal cell irradiation on T cell precursor proliferation. MATERIAL AND METHODS: An in vitro model of thymic stromal cell culture was used. These cultures are able to support early CD7high T cell precursor proliferation and differentiation, mainly towards a CD3+TcRgamma+CD8+ pathway, in the presence of interleukin-7 and stem cell factor. Thymic stromal cell cultures were exposed to 10 Gy 60Co gamma-rays, 24 h before the seeding of CD7high T cell precursors. RESULTS: Irradiation of thymic stromal cells induced a reduction in CD7high T cell precursors, without modifying their differentiation. This effect was reproduced by the addition of supernatants from irradiated stromal cell cultures in sham-irradiated cultures. CONCLUSIONS: These results suggest that gamma-irradiation induces modifications in the production of soluble factors by thymic stromal cells, which in turn modify their ability to support T cell precursor proliferation. To the authors' knowledge this is the first direct demonstration that gamma-irradiation modifies the supportive function of thymic stromal cells. PMID- 9737542 TI - Technical report: Cell thickness measurements by confocal fluorescence microscopy on C3H10T1/2 and V79 cells. AB - Measurements of C3H10T1/2 and V79 cell thickness were performed on living cells by confocal laser fluorescence microscopy. Thickness distributions are reported for cells growing as a monolayer (on mylar and glass) and suspended in their medium. Mean values for cells grown on mylar (corrected for refractive index effects) are 2.9 +/- 0.6 and 6.1 +/- 1.0 microm for C3H10T1/2 and V79 cells respectively. Mean values of the diameters of cells suspended in their medium are 13.0 +/- 1.6 and 9.3 +/- 1.4 microm for C3H10T1/2 and V79 respectively. Knowledge of cell thickness, as irradiated, is of central relevance for studying the relative biological effectiveness of low energy, poorly penetrating radiations. It can be concluded, from the measured cell thickness distributions, that with C3H10T1/2 cells grown on mylar, the LET variation through the whole cell is within 20% for protons and alpha-particles with energies down to 0.6 and 2.5 MeV respectively. From a comparison with thickness values reported in the literature for living or fixed embedded cells growing on plastic substrate, mean values between 2.4 and 3.4 microm and between 6 and 7.5 microm could be assumed for C3H10T1/2 cells and for the most widely used V79 cell lines respectively. PMID- 9737544 TI - Cell death mechanisms in multiple system atrophy. AB - The presence and distribution of apoptotic cell death in multiple system atrophy (MSA) and morphologically related diseases were investigated by means of a modified terminal deoxynucleotidyl transferase-mediated nick end labeling method, comparing their distribution with that of glial cytoplasmic inclusions, immunohistochemically demonstrated bcl-2 protein, bax protein, CD95, TNFalpha, and p53-protein expression, as well as activated microglia. Apoptosis occurred almost exclusively in oligodendrocytes in multiple system atrophy and its general distribution was comparable to the already known oligodendroglial pathology in this disorder. Additionally, in about a quarter of glial cytoplasmic inclusions, there was upregulation of bcl-2-protein and coexpression with ubiquitin, suggesting a final attempt of involved cells to counteract apoptotic cell death. Bax protein was also demonstrated in oligodendroglial cells. A significant neuronal apoptosis was not observed in MSA; these cells might be destroyed secondarily to oligodendroglial apoptosis by necrosis or other forms of programmed cell death. These results emphasize the central role of oligodendroglial pathology in multiple system atrophy, making this disease unique among neurodegenerative diseases. PMID- 9737543 TI - NT-3 attenuates functional and structural disorders in sensory nerves of galactose-fed rats. AB - The present study investigated the effect of NT-3, a neurotrophin expressed in nerve and skeletal muscle, on myelinated fiber disorders of galactose-fed rats. Adult, female Sprague-Dawley rats were fed diets containing complete micronutrient supplements and either 0% D-galactose (control) or 40% D-galactose. Treated controls received 20 mg/kg NT-3 and treated galactose-fed rats received 1, 5, or 20 mg/kg NT-3 three times per week by subcutaneous injections. After 2 months, sciatic and saphenous sensory nerve conduction velocity (SNCV) and sciatic motor nerve conduction velocity (MNCV) were measured and the sciatic, sural, peroneal and saphenous nerves and dorsal and ventral roots processed for light microscopy. Treatment of control animals with NT-3 had no effect on any functional or structural parameter. Compared to control values, galactose feeding induced a sensory and motor nerve conduction deficit and a reduction in axonal caliber. Treatment with 5 and 20 mg/kg NT-3 ameliorated deficits in sciatic and saphenous SNCV in galactose-fed rats but had no effect on the MNCV deficit. NT-3 treatment also attenuated the decrease in mean axonal caliber in the dorsal root and sural nerve but not in the saphenous nerve, ventral root and peroneal nerve. These observations show that NT-3 can selectively attenuate the sensory conduction deficit of galactose neuropathy in a dose-dependent manner that depends only in part on restoration of axonal caliber of large-fiber sensory neurons. PMID- 9737545 TI - Immunohistochemical localization of brain-derived neurotrophic factor in the spinal cords of amyotrophic lateral sclerosis and non-amyotrophic lateral sclerosis patients. AB - Brain-derived neurotrophic factor (BDNF) has a trophic effect on several neuronal subtypes including motor neurons. To localize and assess BDNF in the human spinal cord with particular reference to amyotrophic lateral sclerosis (ALS), we immunohistochemically studied spinal cords from 8 ALS and 13 non-ALS patients. Punctate staining for BDNF was observed in neuronal somata and proximal processes of large-sized anterior horn cells of non-ALS patients, as were distal axons immunolabeled in the neuropil. The same immunostaining pattern was found in the anterior horn cells of ALS patients. Neurons of the dorsal nucleus of Clarke, intermediolateral nucleus, and posterior horn sensory system were also stained in both groups. The results suggest that BDNF may act widely as a trophic factor in the human spinal cord, and motor neurons in ALS patients might be sufficiently supplied with endogenous BDNF from other neuronal subpopulations in the spinal cord. PMID- 9737546 TI - Cell-type-specific enhancement of amyloid-beta deposition in a novel presenilin-1 mutation (P117L). AB - The presenilin-1 (PS1) gene mutation (Pro117Leu), recently identified in a Polish family is characterized by the earliest reported onset (from 24-31 years) of Alzheimer disease (AD) and a very short duration of disease (4-6 years). The neuropathology of 2 subjects with this PS1 mutation (ages at death: 35 and 37 years) was compared to four Down syndrome (DS) patients (mean age at death: 62 years) and 4 sporadic AD patients (mean age at death: 79 years with a mean duration of disease of 18 years). The Polish familial AD (FAD) patients showed a marked increase in the amyloid burden of 2 6-fold in most areas of the brain. The entorhinal cortex was an exception where the amyloid burden was similar in each category of patient. Some brain regions of the Polish FAD patients showed a massive increase of amyloid, such as the molecular layer of the cerebellum where a 7- and 25-fold increase was noted, compared with DS and sporadic AD patients respectively. The cerebellar vessel amyloid burden was also greatly increased in the FAD patients, reflecting a vascular compartment specific increase of amyloid beta deposition. The presence of this PS1 mutation has an even greater effect on both vascular and parenchymal amyloid deposition, than the overexpression of the amyloid beta precursor protein present in DS patients, suggesting that PS mutations can be a critical factor determining amyloid deposition. PMID- 9737547 TI - Expression of matrix metalloproteinases and tissue inhibitors of metalloproteinases in HTLV-I-associated myelopathy. AB - Matrix metalloproteinases (MMPs) have been reported to be involved in inflammatory disorders of the central nervous system (CNS). However, little is known about the role of MMPs in the pathogenesis of HTLV-I-associated myelopathy (HAM)/Tropical spastic paraparesis (TSP). To address this issue, we examined the tissue expression and localization of MMPs and their inhibitors, tissue inhibitors of metalloproteinases (TIMPs) in the spinal cord lesions of HAM/TSP using immunohistochemistry. In addition, the blood and cerebrospinal fluid (CSF) levels of MMPs and TIMPs of the patients with HAM/TSP were determined using sandwich enzyme immunoassays (SIA) and gelatin zymography. Immunohistochemical studies revealed that collagen IV and decorin immunoreactivity on the basement membrane of CNS parenchymal vessels was partially disrupted where inflammatory mononuclear cells infiltrated in active-chronic lesions of HAM/TSP. In these lesions, MMP-2 (gelatinase A) was immunostained mainly on the surface of foamy macrophages and lymphocytes, whereas MMP-9 (gelatinase B) expression was positive in the intravascular and perivascular mononuclear cells but not on foamy macrophages. In contrast, inactive chronic lesions of the spinal cords of the HAM/TSP contained fewer MMP-2-positive or MMP-9-positive mononuclear cells than active-chronic lesions. Many parenchymal vessels had thickened vascular walls which showed increased immunoreactivity to decorin. SIA revealed that production levels of MMP-2 and MMP-9 in both blood and CSF were higher in the patients with HAM/TSP than those in non-inflammatory other neurological disease controls (ONDs). Using zymography, proMMP-9 was detected more frequently in the CSF of patients with HAM/TSP than those in ONDs. Taken together, our data indicate that MMP-2 and MMP-9 may play an important role in the blood-brain barrier breakdown and tissue remodeling in the CNS of HAM/TSP. PMID- 9737548 TI - Increased expression of CNTF receptor alpha in denervated human skeletal muscle. AB - The functional receptor for ciliary neurotrophic factor (CNTF) is comprised of a CNTF binding entity termed CNTF receptor alpha (CNTFRalpha), and 2 signaling molecules called LIF receptor beta and gp130. CNTFRalpha can be released from the cell surface; the soluble form can confer CNTF responsiveness to cells. CNTFRalpha has recently been localized to several nonneuronal cell types including rat skeletal muscle fibers. In this study we examined the expression pattern of CNTFRalpha in normal, denervated and dystrophic human muscle. In muscle biopsies from 12 normal subjects, 16 cases of neurogenic muscular atrophy, 4 cases of Duchenne muscular dystrophy, and 4 cases of limb girdle dystrophy, CNTFRalpha mRNA levels were determined by Northern blotting. Transcript levels were significantly increased in cases of neurogenic atrophy compared to normal controls and dystrophic muscle. By nonradioactive in situ hybridization, CNTFRalpha transcripts were detected in the sarcoplasm of both normal sized and atrophic muscle fibers. In addition, soluble CNTFRalpha was elevated 4.4-fold in the urine of ALS patients compared to normal adults. These results suggest that the expression of CNTFRalpha in human skeletal muscle fibers is regulated by innervation. This regulation appears to be selective, because CNTFRalpha mRNA was not increased in dystrophic human muscle. Increased CNTFRalpha could confer higher sensitivity to CNTF during neurodegeneration or nerve fiber regeneration. PMID- 9737549 TI - Growth factors in infant germinal matrix: relationship to extracellular matrix and cell adhesion molecules. AB - We examined the expression of selected growth factors, growth factor receptors, elements of extracellular matrix and cell adhesion molecules in the germinal matrix layer (GML) utilizing immunohistochemistry and reverse transcriptase polymerase chain reaction. At autopsy brain samples from 10 neonatal infants were used. Epidermal growth factor receptor (EGFR) was significantly expressed in the matrix cells. While transforming growth factor alpha and heparin-binding epidermal growth factor-like growth factor were found in the matrix cells or vascular wall as ligands, epidermal growth factor was not expressed. EGFR and its ligands are thought to be important factors for the maintenance of the matrix cells and cell-to-cell interactions. Insulin like growth factor I, its receptor Ibeta and tenascin were found in the stroma of the GML and periventricular region. Vascular endothelial growth factor and receptor Flk-1, laminin A and B2, fibronectin, collagen type IV and integrins such as beta3, alpha5beta1 and alphaVbeta3 were found mainly in or around the vascular wall indicating their important roles for vascularization. Transforming growth factor beta2 and its receptor II were expressed in the matrix cells and/or vascular wall suggesting a role in proliferation and/or regression of the vasculature. CD44 and Thy-1 were also expressed in the matrix cells. PMID- 9737550 TI - Schwann cell invasion of ventral spinal cord: the effect of irradiation on astrocyte barriers. AB - This study examines a radiation-induced invasion and spread of Schwann cells into ventral gray regions of the lumbar spinal cord. The prevalence of these cells within the gray matter and the time course of their appearance in the ventral spinal cord is quite different from the pattern of Schwann cell development in dorsal spinal cord reported previously. The focus is on 2 possible pathways, each involving astrocytic barriers, by which Schwann cells access the ventral gray matter. The first of these is the glia limitans covering the ventral surface of the spinal cord and the possibility that its integrity has been disrupted by the exposure to x-rays. Comparisons of the glia limitans, including its thickness, between irradiated and nonirradiated rats revealed that exposure to radiation did not result in any morphologically discernible alterations. The second barrier examined was the astrocytic covering of blood vessels. In irradiated animals the astrocyte processes that normally surround blood vessels were missing in some instances, and Schwann cells were observed at these sites. The difference between the dorsal and ventral occurrence of Schwann cells is that, whereas Schwann cells primarily follow axons, specifically dorsal root axons, to access the dorsal spinal cord, it appears that the presence of Schwann cells in the ventral portion of the spinal cord where their location is primarily in the gray matter is associated with the vasculature. PMID- 9737551 TI - Expression of vascular endothelial growth factor (VEGF) and its receptors (Flt-1 and Flk-1) following permanent and transient occlusion of the middle cerebral artery in the rat. AB - Vascular endothelial growth factor (VEGF) is a known endothelial mitogen and a potent enhancer of vascular permeability although its role in focal cerebral ischemia is still not completely understood. The present report describes the immunohistochemical distribution of VEGF and its 2 receptors, Flt-1 and Flk-1 at day 1 and 3 following permanent and transient middle cerebral artery occlusion (MCAO) in the rat. A bilateral increase in VEGF immunoreactivity, particularly in neurons and blood vessels, was seen in both the experimental designs by day 1. By day 3, the immunoreactivity was restricted chiefly to the lesion side, where reaction was most prominent in the border zones of the infarcts. Immunoreaction to VEGF was more pronounced in cases of permanent MCAO than in transient MCAO. Flt-1 reaction was increased in neurons, glial and endothelial cells after both transient and permanent MCAO. Immunoreactivity to Flk-1 was prominent in glial cells and was present to some extent in endothelial cells. These findings indicate an early upregulation of VEGF and its receptors after permanent as well as transient focal cerebral ischemia in the rat. PMID- 9737552 TI - Attributable risk in practice. PMID- 9737553 TI - Attributable fraction for cardiac malformations. AB - To the authors' knowledge, attributable fractions for cardiac malformations have not been reported before. The Baltimore-Washington Infant Study published factors associated with several major cardiac malformations in Maryland, the District of Columbia, and adjacent counties of northern Virginia in 1981-1989. For eight of these malformations, the authors provide attributable fractions of those factors that are potentially causal. Summary attributable fractions range from 13.6% (four factors) for hypoplastic left heart to 30.2% (seven factors) for transposition of great arteries with intact ventricular septum. Extra attributable fraction for factor x, defined as summary attributable fraction for all factors minus that for all but x, is largest for: 1) paternal marijuana use in transposition of great arteries with intact ventricular septum, 7.8%; 2) paternal anesthesia in tetralogy of Fallot, 3.6%; 3) painting in atrioventricular septal defect with Down syndrome, 5.1 %; 4) solvent/degreasing agent exposure in hypoplastic left heart, 4.6%; 5) sympathomimetics in coarctation of aorta, 5.8%; 6) pesticide exposure in isolated membranous ventricular septal defect, 5.5%; 7) hair dye in multiple/multiplex membranous ventricular septal defect, 3.3%; and 8) urinary tract infection in atrial septal defect, 6.4%. Percent-of-cases-exposed dominates relative risk in attributable fraction. If these factors are causal, the larger extra attributable fractions suggest the potential for prevention by specific interventions before/during pregnancy. PMID- 9737554 TI - Population attributable risk of renal cell cancer in Minnesota. AB - A population-based case-control study of renal cell cancer was conducted in Minnesota between 1988 and 1990. It included 690 histologically confirmed incident cases identified through the state cancer surveillance system, and 707 age and sex frequency-matched controls. In this paper, the authors present estimates of the proportion of renal cell cancer cases attributable (or population attributable risk (PAR)) to 1) well-established risk factors, namely smoking, excess weight, and hypertension, and 2) more speculative risk factors, namely elevated protein intake, history of renal disease (i.e., stone, injury, or infection) and high parity among women. These estimates were based on information obtained from directly interviewed subjects (65% of cases and 100% of controls). The PARs for the three main risk factors were 21% for hypertension (defined by a reported personal history of hypertension or of treatment with antihypertensive or diuretic drugs), 21% for excess weight (defined by an elevation of the usual body mass index above the first quartile), and 18% for smoking (past and current). Overall, these three factors accounted for 49% of cases. The proportion increased to 60% when the three more speculative risk factors were considered as well. Sex-specific analyses revealed a greater impact of smoking among men mainly due to a higher prevalence of past smoking. In contrast, the impact of hypertension and excess weight was greater among women due to higher relative risks. These results suggest that 1) intervention measures aimed at reducing smoking, excess weight, and hypertension could substantially lower the overall incidence of renal cell cancer, and 2) intervention measures may have to be sex specific. Conversely, because at least 40% of cases remain unexplained by the risk factors under study, further etiologic research is needed into renal cell cancer, an increasingly common form of cancer. PMID- 9737556 TI - Donation of blood is associated with reduced risk of myocardial infarction. The Kuopio Ischaemic Heart Disease Risk Factor Study. AB - Because high body iron stores have been suggested as a risk factor for acute myocardial infarction, donation of blood could theoretically reduce the risk by lowering body iron stores. For this reason, the authors tested the hypothesis that voluntary blood donation is associated with reduced risk of acute myocardial infarction in a prospective epidemiologic follow-up study in men from eastern Finland. The subjects are all participants of the Kuopio Ischaemic Heart Disease Risk Factor Study. A cohort of 2,862 men aged 42-60 years were followed for an average of almost 9 years. One man (0.7%) out of 153 men who had donated blood in 24 months preceding the baseline examination experienced an acute myocardial infarction during 1984 to 1995, whereas 316 men (12.5%) of 2,529 non-blood donors had an acute myocardial infarction (p < 0.0001 for difference between proportions). In a Cox proportional hazards model adjusting for age, examination years and all other predictive coronary disease risk factors, blood donors had a 88% reduced risk (relative hazard = 0.12, 95% confidence interval 0.02-0.86, p = 0.035) of acute myocardial infarction, compared with non-blood donors. These findings suggest that frequent blood loss through voluntary blood donations may be associated with a reduced risk of acute myocardial infarction in middle-aged men. PMID- 9737555 TI - Effect of change in sodium excretion on change in blood pressure corrected for measurement error. The Trials of Hypertension Prevention, Phase I. AB - Intraperson variability in both blood pressure (BP) and sodium excretion dilutes associations and leads to underestimates of the dose-response relation. The authors applied statistical correction techniques to data from the Trials of Hypertension Prevention (TOHP), Phase I, carried out 1987-1990. Men and women with high normal diastolic BP (80-89 mmHg) were randomized to sodium reduction (n = 327) or a usual care comparison group (n = 417). Regression estimates of the effects of change in sodium and sodium/potassium ratio (Na/K ratio) on blood pressure change in the pooled sample were corrected for both the within-person variance of the excretion measures and the within-person covariance with blood pressure using a multivariate error correction. The estimated cross-sectional reliability was 0.36 for square root(Na) and 0.42 for square root(Na/K ratio) and that for change was 0.31 and 0.28, respectively. Corrected coefficients suggested a decrease of 4.4 mmHg in systolic BP (95% confidence interval (CI) 0.1-8.8) and 2.8 mmHg in diastolic BP (95% CI -0.2 to 5.8) per 100 mmol/24 hour reduction in sodium, and of 3.4 mmHg in systolic BP (95% CI 0.8-6.1) and 1.7 mmHg in diastolic BP (95% CI 0.0-3.5) per unit decrease in Na/K. These results are comparable with those from the Intersalt Study, and suggest that the true effect of sodium change on blood pressure change in normotensives over 18 months is underestimated by more than half in uncorrected data. PMID- 9737557 TI - Influence of consanguinity and maternal education on risk of stillbirth and infant death in Norway, 1967-1993. AB - To analyze the influence of consanguinity and maternal education on stillbirth and infant death for children born in Norway between 1967 and 1993, the authors studied 7,274 children of ethnic Pakistani origin and 1,431,055 children of Norwegian ethnic origin. Of these children, 31.0% of the Pakistani children and 0.1% of the Norwegian children had parents who were first cousins. Consanguinity increased the relative risk of stillbirth (odds ratio (OR) = 1.3, 95% confidence interval (CI) 1.0-1.6) and infant death (OR = 2.4, 95% CI 2.0-3.0), after adjustment for maternal education, maternal age, parity, and year of birth. In 1985-1993, 29% (95% CI 13-45) of stillbirths and infant deaths among the Pakistani group were attributable to consanguinity. In the Norwegian group, 17% (95% CI 13-21) of the deaths were attributable to factors associated with low maternal education, while in the Pakistani group, the corresponding estimate was nonsignificant. The risks of stillbirth and infant death were similar for children with non-consanguineous parents in both populations. This is an important observation considering the differences in socioeconomic status between the two groups. The authors conclude that consanguinity influenced stillbirth and infant death independent of maternal education, and that a large proportion of deaths could be attributed to consanguinity in the Pakistani group due to high frequencies of consanguinity. PMID- 9737558 TI - Associations between family history of asthma, bronchopulmonary dysplasia, and childhood asthma in very low birth weight children. AB - Very low birth weight (VLBW) infants are at risk for childhood wheezing and asthma, as are children with a family history of asthma. Family history of asthma may also be associated with premature labor and, among VLBW infants, with bronchopulmonary dysplasia (BPD) and chronic lung disease (CLD) of prematurity. This study targeted all neonates with birth weight <1,501 g who were admitted to seven perinatal centers in Wisconsin and Iowa between August 1, 1988 and June 30, 1991. Comprehensive information was collected for 723 of the 1,007 30-day survivors, and for 106 full-term controls. A representative subgroup of 257 VLBW children was contacted at age 5 years to ascertain bronchodilator and/or steroid use and diagnosis of asthma. Some evidence of an association between family history of asthma and premature birth was found, but it was not associated with neonatal BPD/CLD or BPD/CLD severity. Among BPD/CLD indicators, radiographic evidence of BPD at age 25-35 days was most strongly associated with bronchodilator use up to age 2 years (odds ratio (OR) = 10.1, 95% confidence interval (CI) 4.07-25.2) and with asthma between ages 2 years and 5 years (OR = 4.83, 95% CI 2.18-10.7). Among children without radiographic evidence of BPD, family history of asthma was associated with childhood asthma and bronchodilator use. PMID- 9737559 TI - Correlates of residential wiring code used in studies of health effects of residential electromagnetic fields. AB - The home wiring code is the most widely used metric for studies of residential electromagnetic field (EMF) exposure and health effects. Despite the fact that wiring code often shows stronger correlations with disease outcome than more direct EMF home assessments, little is known about potential confounders of the wiring code association. In a study carried out in southern Connecticut in 1988 1991, the authors used strict and widely used criteria to assess the wiring codes of 3,259 homes in which respondents lived. They also collected other home characteristics from the tax assessor's office, estimated traffic density around the home from state data, and interviewed each subject (2,967 mothers of reproductive age) for personal characteristics. Women who lived in very high current configuration wiring coded homes were more likely to be in manual jobs and their homes were older (built before 1949, odds ratio (OR) = 73.24, 95% confidence interval (CI) 29.53-181.65) and had lower assessed value and higher traffic densities (highest density quartile, OR = 3.99, 95% CI 1.17-13.62). Because some of these variables have themselves been associated with health outcomes, the possibility of confounding of the wiring code associations must be rigorously evaluated in future EMF research. PMID- 9737560 TI - Use of census-based aggregate variables to proxy for socioeconomic group: evidence from national samples. AB - Increasingly, investigators append census-based socioeconomic characteristics of residential areas to individual records to address the problem of inadequate socioeconomic information on health data sets. Little empirical attention has been given to the validity of this approach. The authors estimate health outcome equations using samples from nationally representative data sets linked to census data. They investigate whether statistical power is sensitive to the timing of census data collection or to the level of aggregation of the census data; whether different census items are conceptually distinct; and whether the use of multiple aggregate measures in health outcome equations improves prediction compared with a single aggregate measure. The authors find little difference in estimates when using 1970 compared with 1980 US Bureau of the Census data or zip code compared with tract level variables. However, aggregate variables are highly multicollinear. Associations of health outcomes with aggregate measures are substantially weaker than with microlevel measures. The authors conclude that aggregate measures can not be interpreted as if they were microlevel variables nor should a specific aggregate measure be interpreted to represent the effects of what it is labeled. PMID- 9737561 TI - Models of survival in HIV infection and their use in the quantification of treatment benefits. AB - Because acquired immunodeficiency syndrome (AIDS) is a shifting endpoint and sufficient follow-up data now allow modeling of survival time (i.e., time from human immunodeficiency virus (HIV) seroconversion to death), the authors evaluated non-parametric and parametric models of mortality with the use of data from 554 seropositive participants in the Vancouver Lymphadenopathy-AIDS Study. The authors then applied these models to quantify treatment benefits at the national level in Canada, using back-calculation and forward-projection based on death registries. The study revealed that the lognormal model better describes survival time than the Weibull model. Relative to observations prior to 1987, later observations (in the era of treatment) revealed a statistically significant change in disease progression: the median survival time increased from 10.1 to 12.0 years, but no further survival improvements were observed in the early 1990s. Concurrent with the increase in availability of treatment, the authors have observed pronounced treatment benefits at the national level: prior to 1995, approximately 1,500 deaths were prevented and 4,200 person-years of life were saved. Also, mortality rates were observed to level off in the mid-1990s due to the shape of the historical HIV infection curve and the accumulating availability of treatment in Canada. PMID- 9737562 TI - Effects of Cryptosporidium parvum infection in Peruvian children: growth faltering and subsequent catch-up growth. AB - The authors conducted a 2-year (1989-1991) community-based longitudinal study in a shantytown in Lima, Peru, to examine the effect of Cryptosporidium parvum infection on child growth during the year following the onset of infection. A cohort of children, aged 0-3 months at recruitment, was followed monthly for anthropometrics, weekly for stool samples, and daily for diarrheal status. Data from 185 children in the cohort permitted a comparison of growth in C. parvum infected and noninfected children. The analyses fitted smooth, flexible curves with a linear random-effects model to estimate growth differences between C. parvum-infected and noninfected children. Children infected with C. parvum experienced growth faltering, both in weight and in height, for several months after the onset of infection, followed by a period of catch-up growth. Younger children took longer to catch up in weight than did older children. Catch-up growth, however, did not occur in children infected between ages 0 and 5 months. These children did not catch up in height, and one year after infection they exhibited an average deficit of 0.95 cm (95% confidence interval (CI) 0.38-1.53) relative to noninfected children of similar age. Stunted children who became infected also did not catch up in either weight or height, and one year after infection they exhibited a height deficit of 1.05 cm (95% CI 0.46-1.66) relative to noninfected, stunted children of similar age. These results indicate that Cryptosporidium parvum has a lasting adverse effect on linear (height) growth, especially when acquired during infancy and when children are stunted before they become infected. PMID- 9737563 TI - Adaptation of a food frequency questionnaire to assess diets of Puerto Rican and non-Hispanic adults. AB - To study issues of diet and health among Hispanic adults living in the northeastern United States, the authors adapted a version of the National Cancer Institute (NCI)/Block food frequency questionnaire. Foods that contributed to nutrient intake of Puerto Rican adults in the Hispanic Health and Nutrition Examination Survey (HHANES) were ranked to identify items to be added to the food list. Portion sizes were compared across HHANES and the Second National Health and Nutrition Examination Survey (NHANES II) to assess the adequacy of the assumed values. Within line items, frequencies of consumption of individual foods were ranked and these data were used to adjust the weighting factors within the database. To test the revised form, 24-hour recalls were collected from 90 elderly Hispanics and 35 elderly non-Hispanic whites. These data were coded into the original and revised food frequency forms and nutrient intake results were compared with recall results by paired t-test, and by Pearson and intraclass correlations. Added foods include plantains, avocado, mango, cassava, empanadas, and custard. Portion sizes differed significantly between HHANES and NHANES II, and were left open-ended. Estimated mean nutrient intakes and correlations with recall data were lower with the original versus the revised form. The authors conclude that the use in minority populations of food frequency questionnaires developed for the general population is likely to result in biased estimates of intake unless modifications are made in the questionnaires. PMID- 9737564 TI - Report of the 1997 Asia Pacific Consensus Conference on the management of Helicobacter pylori infection. AB - While European and United States guidelines for the management of Helicobacter pylori infection have been developed, there are no guidelines for the Asian Pacific. International experts and recognised local authorities met in Singapore in 1997 to develop appropriate guidelines, taking into account the high background prevalence of infection, high incidence rates of gastric cancer and resource limitations. Recommendations were made based on randomised controlled trials or where this was not possible, they were based on the current best available evidence or on good clinical practice. A number of acceptable diagnostic tests for infection are available throughout the region. The non endoscopic methods of choice are the urea breath test or a locally validated antibody test. If endoscopy was to be performed, a biopsy urease test was recommended as the test of first choice, with histology recommended only if this was negative. Post treatment testing was not recommended for all patients; a urea breath test was considered the test of choice if available. All gastric and duodenal ulcer patients who are infected with H. pylori should be treated for H. pylori whether the ulcer is active or in remission. Patients requiring long term non-steroidal anti-inflammatory drug therapy who have a current or recent history of dyspepsia, patients with early gastric cancer or low grade gastric mucosa associated lymphoid tissue lymphoma, and patients with a family history of gastric cancer should be treated. However, it was concluded that there wasn't sufficient evidence that cure of H. pylori infection reduces the risk or prevents the development of gastric adenocarcinoma. Many patients with dyspepsia in the region will request or require early upper endoscopy because of an inherent fear of gastric cancer. However, where endoscopy is not available or is too costly, alternative acceptable approaches were recommended in high risk cancer regions. While evidence is inconclusive to support treatment of H. pylori infection in non ulcer dyspepsia, it was agreed that treatment be offered to patients with documented infection on a case-by-case basis. Treatment regimens need to attain an eradication rate of 90% or greater by per protocol analysis and 80% or greater by intention-to-treat analysis. A number of 7-day regimens were recommended based on available evidence. These regimens were considered likely to maximize the chances of successful eradication with one course of treatment, thereby reducing the risk of acquired antibiotic resistance and leading to long term cost savings. PMID- 9737565 TI - Images in gastroenterology. Duodenal melanosis. PMID- 9737566 TI - Hepatitis viruses under immunosuppressive agents. AB - Clinical and experimental studies have shown that T cell-mediated immune mechanisms are involved in the pathogenesis of hepatitis B virus (HBV) and hepatitis C virus infection. Immunosuppressants may impair T cell function and thereby reduce immune-mediated hepatocytolysis and virus clearance. In addition, corticosteroid may activate the glucocorticoid responsive element in the HBV genome to enhance HBV replication and gene expression. These combined effects result in an increase of viraemia in association with a decrease of serum aminotransferase and hepatic necroinflammation. In acute infection, use of immunosuppressants will increase the incidence of chronic evolution. In chronic infection, withdrawal of immunosuppressants will be followed by a clinical flare due to a rebound of immune attack to hepatocytes with increased viral load. This may lead to a subsequent decrease of the viraemia. Therefore, short-term use of immunosuppressant before antiviral therapy may be beneficial in the treatment of chronic viral hepatitis. However, the clinical rebound may be extremely severe and lead to hepatitis failure; thus, the patients should be monitored closely upon tapering and after the withdrawal of immunosuppressants. Long-term use of immunosuppressants in patients with hepatitis virus infection is usually deleterious, particularly in patients after organ transplantation. These findings suggest that clinicians should be cautious in the use of immunosuppressants in patients with hepatitis virus infection. PMID- 9737567 TI - Helicobacter pylori: from art to a science. AB - Helicobacter pylori infection is widely prevalent especially in developing countries. Increasing knowledge of the pathophysiology associated with H. pylori is leading to an understanding of the mechanisms of mucosal inflammation and gastritis and how this leads to peptic ulcer disease, gastric mucosal associated lymphoid tissues (MALT), lymphoma and gastric cancer. More accurate diagnostic testing for the infection is now possible with both endoscopic and non-endoscopic tests to identify patients most appropriate for eradication therapy. Modern treatments tend to overcome the problems of metronidazole resistance and compliance seen with two week bismuth triple therapy and widely studied is a proton pump inhibitor given with clarithromycin and amoxicillin or metronidazole for one week. These achieve amongst the highest eradication rates and have also been shown to be cost effective. This paper reviews these recent advances and addresses areas of clinical interest and future directions. PMID- 9737568 TI - Cytotoxic T lymphocyte clone specific for autologous human hepatocellular carcinoma cell line SUHC-1. AB - We established a cytotoxic T lymphocyte (CTL) clone directed against an autologous hepatocellular carcinoma (HCC) cell line SUHC-1 which had been established in our department from a patient with HCC associated with hepatitis C virus infection. The CTL clone lysed autologous SUHC-1 cells but did not lyse autologous Epstein-Barr (EB) virus-transformed B cells, natural killer (NK) cell sensitive erythroleukaemia cell line K562, the NK-resistant B cell line Daudi, or allogeneic HCC cell lines, Hep-G2, Hep-3B, Mahlavu and PLC/PRF/5. The CTL clone expressed CD3 and CD8 molecules. The cytotoxic activity of the clone was inhibited by anti-CD3, anti-CD8 and anti-histocompatibility antigen (HLA) class I monoclonal antibodies. These results indicated that the CTL clone recognized HCC tumour antigen in an HLA class I-restricted manner. Furthermore, we investigated the T cell receptor (TCR) gene usage of the CTL clone. The CTL clone expressed TCR alphabeta. We searched for expression of TCR variable (V) alpha and beta regions and sequenced complementary determining region (CDR) 3 of the clone. The clone expressed V alpha14, junctional (J) region alpha9.7 and V beta7, J beta2.1. The amino acid sequence of the N region of the of chain was S-P-G-G-G-G-A-D-G-L-T and of the N-D-N region of the beta chain was S-W-T-G-A-S-T-D-T-Q-Y. These results suggested that HLA class I-restricted CTL play an important role in the elimination of human HCC cells. PMID- 9737569 TI - Combined hepatocellular-cholangiocarcinoma: a clinicopathological study. AB - Combined hepatocellular-cholangiocarcinoma (HCC-CC) is an uncommon form of primary liver cancer having features of both hepatocellular and biliary epithelial differentiation. We reviewed 21 cases of this tumour diagnosed between 1972 and 1996 (patient age range 16-79 years; mean patient age 49.7 years; 18 male and three female patients). Histologically, the majority (n = 18) of tumours were 'mixed' tumours, in which areas of hepatocellular and biliary epithelial differentiation were intimately mixed within the same tumours. Two patients had separate tumours in which discrete nodules of HCC and CC occurred in the same livers. One patient had a 'fibrolamellar' tumour that histologically simulated the fibrolamellar variant of HCC, but some of the tumour cells were mucin producing cells. Of the 21 cases, mucin was demonstrable in 16 and, in the few mucin-negative tumours, electron microscopic studies confirmed the presence of the dual differentiation. The tumours frequently exhibited an invasive character with frequent venous permeation, direct invasion into adjacent liver parenchyma and tumour microsatellite formation, similar to that of ordinary HCC. Histological evidence of cirrhosis or chronic hepatitis was present in 77.8% of patients and 75% of patients were hepatitis B surface antigen positive. Raised serum alpha-fetoprotein (AFP) levels (above 300 ng/mL) were present in 61.5% of patients and AFP was detected immunohistochemically in 55% of tumours. The overall survival times of patients with HCC-CC were short. In conclusion, HCC-CC showed clinical and pathological features more akin to those of ordinary HCC than to CC. PMID- 9737570 TI - Intra-arterial carbon dioxide-enhanced ultrasonogram of hepatocellular carcinoma treated by transcatheter arterial embolization and percutaneous ethanol injection therapy. AB - The purpose of this study was to investigate the value of carbon dioxide-enhanced ultrasonography (CO2-US) in the evaluation of viable hepatocellular carcinomas (HCC) which were treated by transcatheter arterial embolization (TAE), percutaneous ethanol injection (PEI), or a combination treatment (TAE and PEI). Forty-one patients with 66 HCC were included in the study. They underwent CO2-US and angiography were performed in all tumours after they were treated by TAE, PEI or a combination treatment. Forty-six tumours were positively enhanced by CO2-US and 40 of them were positive by angiography. These 46 tumours were proved to be viable tumours either by biopsy or by follow-up studies. The positive predictive value was 100% for CO2-US and 87.8% in angiography. Twenty tumours were negative by CO2-US and these were also negative by angiography. Carbon dioxide-enhanced ultrasonography is a more reliable method for detecting the viable portion of the treated HCC compared with conventional angiography. PMID- 9737571 TI - Case report: Insulin-like growth factor II expression in hepatocellular carcinoma with alcoholic liver fibrosis accompanied by hypoglycaemia. AB - This case of hepatocellular carcinoma (HCC) with alcoholic liver fibrosis, which was not associated with hepatitis viruses, was accompanied by hypoglycaemia. The immunoreactive insulin level was low and other hormonal examinations were almost normal. Immunohistochemical studies showed a high level of insulin-like growth factor II (IGF2) peptide in the HCC section and the size heterogeneity of serum IGF2 investigated by western blot revealed a large form at approximately 15 kDa. These results suggest that the HCC with alcoholic liver fibrosis produced IGF2 and that the hypoglycaemia was caused by tumour-associated IGF2. PMID- 9737572 TI - Histological changes of concurrent hepatitis C virus infection in asymptomatic hepatitis B virus patients. AB - In chimpanzees and in vitro cell culture studies, hepatitis C infection has been shown to suppress hepatitis B virus expression. In addition, hepatitis C infection can cause much more severe liver disease in patients chronically infected with hepatitis B virus. The aims of the present study were to determine the prevalence of hepatitis C infection in asymptomatic chronic hepatitis B Hong Kong Chinese patients and the histological changes and hepatic expression of hepatitis B virus and hepatitis C virus. Five hundred and seventy-one Hong Kong Chinese asymptomatic chronic hepatitis B patients were studied. Only four (0.7%) were hepatitis C virus antibody positive; they were also all positive for hepatitis C viral RNA in serum by reverse transcription-polymerase chain reaction (RT-PCR). Portal lymphoid aggregates and bile duct damage was noted in the liver sections of three of the four patients. Hepatic expression of hepatitis B surface antigen was detected in three patients; none had detectable hepatitis B core antigen. By branched DNA assay, serum hepatitis B DNA could not be detected in any of the four patients, but three had hepatitis C RNA. By in situ RT-PCR, hepatitis C RNA was detected in the cytoplasm of three of the four patients. These findings suggest that hepatitis C coinfection in asymptomatic chronic hepatitis B patients is uncommon in Hong Kong Chinese and active hepatitis B viral replication is absent in these patients. PMID- 9737573 TI - Diagnostic value of anti-hepatitis D virus (HDV) antibodies revisited: a study of total and IgM anti-HDV compared with detection of HDV-RNA by polymerase chain reaction. AB - A high serum titre (> or = 1000 or > or = 5000) of total antibody to hepatitis D virus (anti-HDV) and positive for immunoglobulin (Ig)M anti-HDV have been used to represent HDV replication, while reverse transcription-polymerase chain reaction (RT-PCR) is currently the most sensitive assay for detecting HDV viraemia. The aim of the present study was to re-evaluate the correlation of total anti-HDV and IgM anti-HDV with HDV viraemia based on RT-PCR and to assess the clinical significance of these markers in acute and chronic HDV superinfection. Chronic HDV infection was defined as positive HDV-RNA by RT-PCR for more than 6 months, while total anti-HDV titre was defined by serial dilution. Of 178 hepatitis B virus (HBV) carrier patients studied, 119 cases had been anti-HDV positive for more than 6 months. Two-thirds (79/119) were positive for HDV viraemia by RT-PCR. Only half the chronic HDV viraemic patients had a high titre (> or = 1000) of total anti-HDV, and there was only moderate agreement (kappa = 0.41) between total anti-HDV titre/IgM anti-HDV and HDV-RNA and chronic HDV viraemia. Based on cross-sectional and longitudinal follow-up analyses, serum total anti-HDV titres > or = 100 appeared to be an excellent cut-off titre (kappa = 0.91) in differentiating chronic from acute HDV infection among viraemic patients. In summary, IgM and a high titre total of anti-HDV are not good markers of HDV viraemia, but an anti-HDV titre of > or = 100 appears to be an excellent marker for the differentiation of acute from chronic HDV superinfection. PMID- 9737574 TI - Acute hepatitis C among renal failure patients on chronic haemodialysis. AB - Hepatitis C virus (HCV) infection is common in haemodialysis units, yet little information is available about the clinical feature of acute hepatitis C among renal failure patients. The present study is based on 49 cases of acute hepatitis C seen at a haemodialysis centre where sporadic nosocomial infection was occurring up to June 1993. Liver function tests were done at 4 weekly intervals on all dialysis patients, anti-HCV antibodies were tested by the C-100 and second generation tests and serum HCV-RNA was determined by the branched DNA and Amplicore tests. Diagnosis of acute hepatitis C was made on the basis of an acute rise in alanine aminotransferase (ALT) and seroconversion to positive anti-HCV antibodies. Clinical presentation of acute hepatitis was generally mild with rare overt jaundice and the diagnosis was possible only from increased ALT, which was generally low. Spontaneous resolution of acute hepatitis within 8 months with clearance of viral RNA occurred in only four cases, 91.8% of patients developing chronic hepatitis. Biopsy in 12 cases with high ALT levels showed mild to moderate inflammatory activities. In conclusion, the clinical presentation of acute hepatitis C is generally mild in chronic haemodialysis patients, but spontaneous resolution is infrequent. A longer follow-up period is required for defining the long-term prognosis. PMID- 9737575 TI - Clinical significance of serum hyaluronic acid as a fibrosis marker in chronic hepatitis C patients treated with interferon-alpha: histological evaluation by a modified histological activity index scoring system. AB - The aim of the present study was to investigate the histological changes effected by interferon (IFN) treatment and to evaluate the clinical significance of serum hyaluronic acid (HA) as a marker of fibrosis. Forty-nine patients with chronic hepatitis C treated with IFN-alpha were divided into three groups according to the existence of viraemia: sustained complete responders (CR), complete responders with relapse (PR) and non-responders (NR). Needle biopsy sections of the liver taken before and at the end of IFN treatment were assessed according to the modified histological activity index (HAI) scoring system. Serum fibrosis markers, including HA, were measured at needle biopsies. Biopsies of CR at the end of treatment showed a significant improvement in fibrosis and necroinflammatory scores. More significant correlation was observed between fibrosis scores and serum levels of HA before IFN treatment (r = 0.607, P < 0.0001) than those between fibrosis scores, on the one hand, and peptide of type III procollagen (PIIIP; r = 0.531, P = 0.0004) or type IV collagen 7S domain (type IV-C; r = 0.241, P = 0.1062) on the other. Moreover, serum HA levels fell significantly in patients in whom fibrosis improved (P = 0.011). This is the first paper describing the advantages of the modified HAI scoring system over others in estimating the effect of IFN-alpha; the results also indicate that serum HA can be useful in monitoring liver fibrosis in chronic hepatitis C patients treated with IFN-alpha. PMID- 9737576 TI - Computer morphometry for quantitative measurement of liver fibrosis: comparison with Knodell's score, colorimetry and conventional description reports. AB - Liver fibrosis is currently described quite subjectively or, at best, semiquantitatively by scoring systems. In order to measure the severity of liver fibrosis quantitatively and to compare this with established methods, such as Knodell's scoring system, the colorimetric method and conventional description reports, we undertook the present study. A personal computer with an image grabber card and a microscope equipped with a computer-controlled slide-driver was used for computer morphometry. The principle behind morphometry is based on the different colours of hepatocytes and fibres following staining with Masson's trichrome stain. There were 31 patients (25 male, six female) recruited into the present study with a mean +/- SD age of 41.6 +/- 15 years (range 24-66 years). Of these patients, 16 had chronic hepatitis B, 12 had chronic hepatitis C and three were alcoholics. Colorimetric methods and Knodell's fibrosis score were performed according to established protocols. Conventional description reports were obtained from reviews of patient charts. The results from computer morphometry were highly correlated with results from the colorimetric method, with a correlation coefficiency gamma = 0.85 (P<0.0001). The results from computer morphometry also correlated with both Knodell's scoring system (gamma = 0.69; P<0.001) and conventional description reports (gamma = 0.46; P<0.01). Results from Knodell's scoring system were significantly correlated with computer morphometry, as follows: score 0, 2.7 +/- 1.4; score 1, 5.7 +/- 1.2; score 2, 7.7 +/- 2.3; score 3, 10.7 +/- 3.2; score 4, 21.8 +/- 14.1. The trend was statistically significant by the Wilcoxon rank sum test. In conclusion, our computerized morphometry system is a reliable tool for the evaluation of the severity of liver fibrosis and can be used as a tool for the objective quantification of liver fibrosis. PMID- 9737577 TI - Bile acids influence hepatic chemiluminescence in normal and oxidative-stressed rats. AB - The aim of the present study was to clarify whether bile acids influence chemiluminescence (CL) in the liver in vivo. Hepatic CL was determined on the surface of the liver of anaesthetized rats by using a photon counter. In normal rats, hepatic CL was significantly decreased 30 min after enteral administration of chenodeoxycholic acid (CDCA) or deoxycholic acid (DCA), but returned to its initial level 3 h later, after part of the CDCA administered was metabolized. Ursodeoxycholic acid (UDCA) and cholic acid had no effect on CL. In contrast, hepatic CL was markedly increased 30 min after CDCA or DCA administration in rats given either buthionine sulphoximine (BSO), an inhibitor of gamma glutamylcysteine synthetase, or diethyldithiocarbamate (DDC), an inhibitor of both superoxide dismutase and glutathione peroxidase. Chenodeoxycholic acid further increased the CL of BSO- or DDC-treated rats during inhalation of oxygen via a tracheal cannula. Coadministration of UDCA eliminated the effects of CDCA on the hepatic CL of normal and BSO- or DDC-treated rats with or without oxygen inhalation. We conclude that cytotoxic bile acids, such as CDCA, increase CL in the antioxidants-depleted or oxidative-stressed liver in vivo, but that UDCA prevents CDCA from developing CL. PMID- 9737578 TI - Regulation of uridine diphosphate glucuronosyltransferase during the acute-phase response. AB - The acute-phase response is associated with profound effects on oxidative drug metabolism. However, the effects on glucuronidation are poorly characterized. The aim of the present study was to determine the role of mediators of the acute phase response in the regulation of hepatic uridine diphosphate glucuronosyltransferase (UGT) expression. Family 1 and family 2 UGT isoforms were studied in turpentine-injected rats and in primary hepatocyte cultures exposed to cytokines and/or dexamethasone. In the in vivo model, glucuronidation of p nitrophenol was unaffected, while testosterone glucuronidation was reduced to 65% of control (P<0.01). In contrast, the mRNA level of UGT1*1 (which metabolizes bilirubin, not phenols) was depressed to 16% of control (P<0.002), while the mRNA level of UGT2B3 (which metabolizes testosterone) was reduced to 53% (P<0.05). In primary hepatocyte culture, dexamethasone treatment resulted in a 3.4-fold induction of UGT1*1 mRNA levels (P<0.001) but only a 1.5-fold induction of UGT2B3 (P=0.1). Interleukin-6 in the presence of dexamethasone resulted in a marked dose dependent suppression of both UGT1*1 and UGT2B3, although to different degrees. Interleukin-1 had no effect on UGT mRNA levels. Thus, inflammatory mediators, such as cytokines and glucocorticoids, may be important determinants of both oxidative and conjugative drug metabolism by the liver. PMID- 9737579 TI - Gastrin release and gastric acid secretion in the rat infected with either Helicobacter felis or Helicobacter heilmannii. AB - Helicobacter pylori infection in humans has been shown to be associated with changes in gastric physiology, including exaggerated basal and meal-stimulated gastrin levels. This has been suggested to be due to the direct effects of the bacterium through inflammation and its urease enzyme. The gastric bacteria Helicobacter felis and Helicobacter heilmannii colonize the antrum of rats in large numbers and induce no significant inflammatory response. Thus, the direct effect of Helicobacter infection on gastric physiology, independent of gastritis, could be studied. Basal, freely fed and stimulated acid and gastrin levels were recorded from animals infected with H. felis, H. heilmannii or uninfected controls over a 30 week period. No significant difference was found between freely fed gastrin over 7 weeks or fasting gastrin over 24 weeks or basal and stimulated acid over 30 weeks between all three groups. Triple therapy did not alter gastrin or acid output. The antrum of all Helicobacter-infected rats was well colonized; triple therapy cleared H. felis but not H. heilmannii. Very little inflammation was seen in control or Helicobacter-infected animals. In conclusion, Helicobacter-induced effects on gastric physiology are unlikely to be due to direct bacterial effects, but are best explained by other factors (i.e. inflammatory damage). PMID- 9737580 TI - A protease inhibitor attenuates gastric erosions and microcirculatory disturbance in the early period after thermal injury in rats. AB - The effects of camostat mesilate, a synthetic serine protease inhibitor on gastric microcirculation and active oxygen species generated by leucocytes from the gastric and jugular veins in the early period after thermal injury were assessed. Male Wistar rats were anaesthetized and a 30% full skin-thickness dorsal burn was inflicted. Camostat mesilate (100 mg/kg) was dissolved in distilled water and administered orally to rats 40 min before thermal injury (the camostat group). The control animals (the vehicle group) were administered distilled water orally. Rolling leucocytes as well as Monastral blue B deposits in venules were observed using in vivo microscopy. Active oxygen species were measured by chemiluminescence. Camostat mesilate decreased the total length of gastric erosion, venular deposits of Monastral blue B, and rolling of leucocytes in venules, and relatively increased luminol-dependent chemiluminescence activity generated by zymosan-stimulated leucocytes 15 min after thermal injury. These results suggest that serine proteases are involved in the formation of gastric erosions and gastric microcirculatory disturbance in the early period after thermal injury. PMID- 9737581 TI - Spontaneous bacterial peritonitis in fulminant hepatic failure. AB - Ascites may be associated with fulminant hepatic failure (FHF), but spontaneous bacterial peritonitis (SBP) is an extremely rare complication. We report on two patients with FHF who developed SBP. One patient died and the other recovered. PMID- 9737582 TI - The role of dendritic cells in the infection of CD4+ T cells with the human immunodeficiency virus: use of dendritic cells from individuals homozygous for the delta32CCR5 allele as a model. AB - Despite exposure to multiple strains of both macrophage (M)-tropic and T cell (T) tropic HIV, primary infection is largely restricted to relatively homogeneous M tropic virus. Since dendritic cells (DCs) play a pivotal role in the early events of HIV infection, several studies have focused on the role of DCs in this restriction. It has been proposed that DCs are more efficiently infected with M tropic versus T-tropic viruses; however, the infectability of DCs and the relevance of their infectability for inducing productive infection is controversial. It has also been suggested that variability in DC expression of coreceptors for M-tropic versus T-tropic virus could explain the restriction in the transmitting virus. Using HIV-pulsed DCs from individuals with a homozygous deletion in the CCR5 gene as a human "knockout" model, we demonstrate that infection of DCs per se is not necessary to promulgate infection in CD4+ T cells. The data also suggest that transmission of HIV to CD4+ T cells is not dependent on DC coreceptor expression. PMID- 9737583 TI - Molecular characterization of five neutralizing anti-HIV type 1 antibodies: identification of nonconventional D segments in the human monoclonal antibodies 2G12 and 2F5. AB - We have stabilized a panel of 33 hybridomas producing human monoclonal antibodies (MAbs) against HIV-1 gp160 and p24. Five of these antibodies were able to neutralize different HIV-1 isolates, and two of them (2F5 and 2G12) revealed remarkable potential to neutralize primary virus isolates of different clades in several in vitro tests. To determine whether a structural basis for neutralization could be identified, we analyzed the antibodies at the molecular level. This study reports the primary nucleotide and deduced amino acid sequences of the rearranged heavy and light chain V segments (VH, Vkappa) of the neutralizing MAbs (1B1, 1F7, 2F5, 2G12, and 3D5) and the nonneutralizing anti gp41 MAb 3D6. Aligning the V segments with the nearest related germline genes illustrated the occurrence of somatic mutations. The neutralizing MAbs show mutational rates comparable to those of antibodies that appear in patients in whom the immune system is under constant antigenic pressure over a long period of time. In contrast, 3D6, which recognizes the immunodominant region on gp41, displays homologies as high as 97 and 98% compared with its VH and Vkappa germline genes. The diversity segments [D(H)] of 1B1, 1F7, 3D5, and 3D6 were assigned to single D(H) segments on the chromosomal D(H) locus. 2F5 presents a D(H) segment 52 nucleotides in length, which could be explained by fusion of two segments on the immunoglobulin heavy chain locus that have not yet been described as rearranged regions. 2G12 D(H) shows best homologies to a D(H) segment between D3-22 and D4-23. This D(H) segment could be the reason for the rare occurrence of antibodies competing with 2G12. Since this nearest related chromosomal region on the D(H) locus does not display recombination signals at the flanking regions, this segment is normally not taken into consideration as a site for immunoglobulin heavy chain rearrangement. PMID- 9737584 TI - Effect of major deletions in the V1 and V2 loops of a macrophage-tropic HIV type 1 isolate on viral envelope structure, cell entry, and replication. AB - Two HIV-1 envelope mutant proteins were generated by introducing deletions in the first and second hypervariable gp120 regions (V1 and V2 loops, respectively) of a macrophage-tropic primary HIV-1 isolate, SF162, to study the effect of the deleted sequences on envelope structure, viral entry, and replication potentials. The first mutant lacked 17 amino acids of the V1 loop and the latter 30 amino acids of the V2 loop. A comparison of the immunochemical structure of the wild type and mutant monomeric and virion-associated gp120 molecules revealed that the V1 and V2 loop deletions differentially altered the structure of the V3 loop, the CD4-binding site, and epitopes within conserved regions of gp120. Regardless of differences in structure, both mutated envelope proteins supported viral replication into peripheral blood mononuclear cells to levels comparable to those of the wild-type SF162 virus. However, they decreased the viral replication potential in macrophages, even though they did not alter the coreceptor usage of the viruses. These studies support and extend previous observations that a complex structural interaction between the V1, V2, and V3 loops and elements of the CD4-binding site of gp120 controls entry of virus into cells. The present studies, however, suggest that the effect of the V1 and V2 loops in viral entry is cell dependent. PMID- 9737585 TI - Cell-associated viral RNA expression during acute infection with HIV type 1. AB - The mechanism of decline in viremia following acute infection with HIV is unknown. To characterize this process virologically, the expression of viral RNAs was analyzed in samples of peripheral blood mononuclear cells (PBMCs) from a patient who experienced a 100-fold decline in plasma viremia over a 13-day period prior to the initiation of antiretroviral therapy. Cell-associated viral RNA declined in association with the decline in plasma virus. During the initial 7 days of observation, plasma viremia declined more than 10-fold with no change in the ratio of unspliced to multiply spliced mRNAs. The efficiency of viral gene expression did not decline during the study period and varied from 380 to 2800 unspliced RNA copies per productively infected cell. Together, these data indicate no change in the relative proportion of cells in late- and early-stage gene expression during the initial decline and provide evidence against shortening of the viral replication cycle by immune surveillance. However, the prevalence of productively infected cells declined markedly during the 13 days of observation, from 1 in 250 to 1 in 25,000 PBMCs. These data are compatible with depletion of available target cells during the initial decline in viremia. As the level of plasma virus stabilized after 8 days of observation, the ratio of unspliced to multiply spliced mRNAs rose; this rise was due to a relatively greater decline in multiply spliced mRNA. These data suggest the possible onset of a blockade to new infection events (for example, by neutralizing antibody or chemokines), causing an increase in the relative proportion of cells in late stage gene expression. They may also be explained, however, by the persistence of cell-associated virions together with the near disappearance of productively infected cells from the circulation. PMID- 9737586 TI - Optimization of polymerase chain reaction for detection of HIV type 2 DNA. AB - The aim of this study was to increase the sensitivity of an earlier version of an HIV-2 nested PCR assay based on primers in the gag, pol, LTR, and env regions. The assay was first optimized with regard to concentrations of dNTP, MgCl2, and primers, using a method that allowed optimization of all three parameters in only two test runs. We then designed and optimized new primer sets in the LTR, gag, and gag/pol regions that were based on more isolates than were the former (old) primer sets. Samples from 57 HIV-2 antibody-positive individuals were tested with the four old primer sets as well as with the three new primer sets. Five primer sets from this run (new gag, new gag/pol, old LTR, old env, and new LTR) were then tested with 35 more samples, giving a total number of 92 tested samples from HIV-2-infected individuals. At initial testing of the 92 samples a combination of primer sets from two different regions yielded a sensitivity ranging from 93.5 to 98.9%. After repeated testing the sensitivity ranged from 96.7 to 100% for the different primer combinations. The specificity was 100% for all primer sets except old LTR, which had a specificity of 97%. In conclusion, it is possible to create a more sensitive PCR assay by optimizing the different PCR parameters as well as by including primer sets based on more isolates. PMID- 9737587 TI - Antibodies to the HIV type 2 core protein p26 and Vpx: association with disease progression. AB - A longitudinal cohort study was conducted to define the prevalence and temporal pattern of antibody response to the HIV-2 virion-associated proteins p26gag and Vpx. One hundred and forty-one asymptomatic HIV-2-infected women were enrolled, and followed for up to 11 years. Eighty-one percent of the subjects had antibodies to p26, and 51% to Vpx; response to these two antigens was not correlated. The response to both proteins was determined early in infection, and remained stable over time. The absence of antibodies to p26 was a highly significant predictor of CDC category IV HIV-related disease (p < 0.01) in both univariate and multivariate analysis. Antibody response to Vpx alone was not associated with disease progression. However, those individuals lacking anti-p26 antibodies, and with anti-Vpx antibodies, were six times more likely to be classified as CDC category IV by the end of the study (p < 0.01). This represents the first identification of virus-specific serological markers for HIV-2-related disease progression. PMID- 9737588 TI - SIV-associated nephropathy in rhesus macaques infected with lymphocyte-tropic SIVmac239. AB - We examined the renal pathology and viral genetic changes following inoculation of six rhesus macaques with lymphocyte-tropic SIVmac239. Portions of the renal cortex were sieved into glomerular and tubulointerstitial (TI) fractions and examined for SIVmac sequences by PCR and for p27 core antigen. SIVmac sequences were detected in renal tissue from five of six macaques (three of five glomerular and five of five TI fractions were positive for SIV by PCR). Glomerulosclerosis (segmental and global) was evident in two macaques that were positive for env sequences in the glomerular fractions. Diffuse mesangial hyperplasia and matrix expansion were present in all three animals with glomerular SIV, as was an increase in glomerular collagen I and collagen IV. Tubulointerstitial inflammation was evident in all virus-inoculated macaques. The TI infiltration of CD68+ cells was most pronounced in the animals with SIVmac present in the glomerulus. All SIVmac-infected macaques exhibited increased glomerular deposition of IgM and to a lesser extent IgG, but no C3 or IgA was evident. Sequence analyses of the viral env gene (gp120) isolated from the glomerular and TI fractions of a macaque that developed glomerulopathy revealed the presence of specific viral variants in glomerular and TI fractions. In addition, chimeric viruses constructed with glomerular but not tubulointerstitial gp120 sequences were converted to a macrophage-tropic phenotype. These results indicate that infection by lymphocyte-tropic SIVmac239 is primarily associated with immunoglobulin deposition in the glomerulus and suggests that when glomerulosclerosis develops there is selection of viral variants that are macrophage tropic in nature. PMID- 9737589 TI - Phenotypic changes in peripheral blood monocytes of cynomolgus monkeys acutely infected with simian immunodeficiency virus. AB - The quantitative and phenotypic changes of peripheral blood monocytes during the acute stage of simian immunodeficiency virus infection were investigated. We inoculated intravenously three cynomolgus monkeys (Macaca fascicularis) with 100 TCID50 of SIVmac239 and collected whole blood twice a week until 35 days postinoculation. We found that the relative number of monocytes in peripheral blood leukocytes significantly increased at 7-17 days postinoculation. This increase was concomitant with the peak of primary SIV antigenemia. To determine if the monocytes observed during the acute stage were phenotypically altered, they were periodically examined for the expression of surface markers (i.e., CD11b, CD14, CD16, CD29, D32, CD56, CD62L, CD64, CD80, and MHC-II-DR) by flow cytometry. The results showed that the expression levels of CD14 and CD56 on most of the monocytes were remarkably reduced at 7-17 days postinoculation, and a new subpopulation, CD14lowCD16+CD80+ monocytes, was clearly detected at 10 days postinoculation. These results indicate that the phenotypic alteration of peripheral blood monocytes occurs during the primary SIV infection. PMID- 9737590 TI - Mechanisms of protection induced by attenuated simian immunodeficiency virus. II. Lymphocyte depletion does not abrogate protection. AB - To determine the role that cellular immune responses play in the protection conferred by vaccination with attenuated SIVmac32H (pC8), we have attempted to deplete macaques of their CD8+ cells prior to challenge with wild-type SIVmac32H (pJ5). In two of four pC8-infected macaques, N109 and N112, a transient partial depletion of CD8+ cells by antibody treatment was achieved. On the day of challenge peripheral CD2+CD4-CD8+ cell counts were reduced by 92 and 95%, respectively, in animals N109 and N112 and their lymph nodes revealed a 46 and 58% reduction, respectively, in CD2+CD4-CD8+ cells. Two other pC8-immunized macaques, N110 and N111, treated in the same way, did not show significant depletion of CD8+ cells. None of these four pC8-immunized animals became infected when challenged with 50 MID50 of pJ5. Treatment of a further four pC8-infected and protected macaques and two naive control animals with Campath-1H antibody successfully depleted peripheral CD3+ cell counts by >99% in all treated animals. Campath-1H depletion resulted in enhanced, longer lasting lymphoid depletion. Yet subsequent challenge with 20 MID50 of pJ5 still failed to infect the pC8 immunized animals. All eight of the naive controls, including two Campath-1H treated animals, became infected following challenge. In summary, partial depletion of circulating CD8+ cells or total lymphocytes prior to challenge failed to abrogate the protection conferred by vaccination with pC8. PMID- 9737591 TI - Sequence analysis of the first HTLV-I infection in Germany without relations to endemic areas. AB - In most parts of Europe only a limited number of sporadic cases of HTLV-I infections have been identified. So far, the few cases found in Germany have always been linked to individuals with relations to endemic areas. Here we report the first HTLV-I infection from a German ATL patient without any known risk for HTLV-I infection and with no relations to known endemic areas. The DNA sequence of the provirus was determined, and a phylogenetic analysis based on the LTR sequence established a close relationship with HTLV-I sequences previously found in two Romanian patients. Our data suggest the existence of a previously unrecognized cluster of HTLV-I infections in southeastern or central Europe. PMID- 9737592 TI - Molecular epidemiological analysis of human immunodeficiency virus type 1 in Newfoundland, Canada. PMID- 9737593 TI - Resistance management strategies in malaria vector mosquito control. Baseline data for a large-scale field trial against Anopheles albimanus in Mexico. AB - A high level of DDT resistance and low levels of resistance to organophosphorus, carbamate and pyrethroid insecticides were detected by discriminating dose assays in field populations of Anopheles albimanus in Chiapas, southern Mexico, prior to a large-scale resistance management project described by Hemingway et al. (1997). Biochemical assays showed that the DDT resistance was caused by elevated levels of glutathione S-transferase (GST) activity leading to increased rates of metabolism of DDT to DDE. The numbers of individuals with elevated GST and DDT resistance were well correlated, suggesting that this is the only major DDT resistance mechanism in this population. The carbamate resistance in this population is conferred by an altered acetylcholinesterase (AChE)-based resistance mechanism. The level of resistance observed in the bioassays correlates with the frequency of individuals homozygous for the altered AChE allele. This suggests that the level of resistance conferred by this mechanism in its heterozygous state is below the level of detection by the WHO carbamate discriminating dosage bioassay. The low levels of organophosphate (OP) and pyrethroid resistance could be conferred by either the elevated esterase or monooxygenase enzymes. The esterases were elevated only with the substrate pNPA, and are unlikely to be causing broad spectrum OP resistance. The altered AChE mechanism may also be contributing to the OP but not the pyrethroid resistance. Significant differences in resistance gene frequencies were obtained from the F1 mosquitoes resulting from adults obtained by different collection methods. This may be caused by different insecticide selection pressures on the insects immediately prior to collection, or may be an indication that the indoor- and outdoor-resting A. albimanus collections are not from a randomly mating single population. The underlying genetic variability of the populations is currently being investigated by molecular methods. PMID- 9737595 TI - Use of odour-baited sticky boards to trap tabanid flies and investigate repellents. AB - Tabanid flies were captured in a sheep pasture in Hungary using black plastic boards (30 x 30 cm), coated with an adhesive sheet, which were either unbaited or baited with ethane-ethiol or Swormlure-4. Five species of tabanid were caught, of which Tabanus tergestinus was caught in the greatest number (87% of 192). The sex ratio of T. tergestinus on unbaited boards was not statistically significantly different to equality. However, on baited boards, males were significantly more numerous, probably due to a repellent effect of the odours on female tabanids. Because of the importance of repellents in tabanid control, this result encourages the exploration of a wider range of compounds as tabanid repellents than are used at present. Sticky boards offer a simple technique for trapping both sexes of tabanids, especially males, which are poorly represented in many tabanid traps. PMID- 9737594 TI - A survey of the ixodid ticks parasitising cattle in the Eastern province of Zambia. AB - The results of tick surveys carried out in the Eastern province of Zambia between December 1982 and February 1996 were principally in agreement with the findings of earlier surveys conducted during the period 1965-72. Boophilus decoloratus has almost been replaced by Boophilus microplus. Hyalomma truncatum was found in small numbers throughout the province and Hyalomma marginatum rufipes was only rarely encountered in collections made from cattle. Rhipicephalus evertsi evertsi was largely confined to the southern part of the plateau and the valleys of the Zambezi tributaries. The main difference between the present survey and the previous one concerns the status of Rhipicephalus appendiculatus. This species is currently expanding its range in a westward direction, whereas it was virtually absent from the southern part of the province during the period 1965-72. The majority of specimens collected are morphologically intermediate between R. appendiculatus and the closely allied Rhipicephalus zambeziensis. The available evidence indicates that R. zambeziensis (sensu stricto) is absent from the province. The phenology of R. appendiculatus is aberrant in the province: at lower altitudes a second generation of adult ticks is recorded on the hosts at the start of the dry season. PMID- 9737596 TI - A new species of the Simulium damnosum complex from Uganda, and comparative morphology of the tarsal claws in females of this complex. AB - The female, male, pupa and larva of Simulium (Edwardsellum) pandanophilum sp. nov. (Diptera: Simuliidae) from western Uganda are described. Diagnostic features are the shape of the tarsal claws of females and, cytogenetically, new intraspecific inversions on chromosomes I and II. Simulium pandanophilum is assigned to the Simulium damnosum complex. Different tarsal claw shapes of West African members of the S. damnosum complex are also described. PMID- 9737597 TI - Use of climatic data and satellite imagery to model the abundance of Culicoides imicola, the vector of African horse sickness virus, in Morocco. AB - African horse sickness (AHS) is a vector-borne, infectious disease of equids caused by African horse sickness virus (AHSV). The only proven field vector of the virus is the biting midge Culicoides imicola. Following a recent epizootic (1989-91) of AHS in Morocco, light traps and automatic weather stations were operated for 2 years at twenty-two sites distributed over much of the country. The annually-averaged mean daily trap catch of C. imicola at these sites was negatively correlated with wind speed, and positively correlated with the average and mean annual minimum NDVI (Normalized Difference Vegetation Index, a remotely sensed measure of vegetation activity). There were no significant correlations between the mean daily trap catch and air temperature, soil temperature, relative humidity, saturation deficit, rainfall, altitude or the mean annual maximum or range of NDVI. The best two-variable model, which combined WindspeedMnAvMn (the average daily minimum wind speed of the least windy month) and NDVImin (the average annual minimum NDVI) as predictors, explained over 50% of the variance in the annually-averaged mean daily trap catch of C. imicola. There was a significant, positive correlation between minimum wind speed at night and the daily mortality rate of adult female C. imicola and it is suggested that the relationship between wind speed and the abundance of C. imicola arises from effects on adult mortality or dispersal. Considering several climatic variables, in North Africa NDVImin was most significantly correlated with total annual rainfall. It is suggested that the relationship between NDVImin and the abundance of C. imicola arises from the impact of soil moisture on both. It is proposed that areas of Morocco with higher levels of soil moisture in late summer or autumn provide more, larger and/or more enduring breeding sites for C. imicola, as well as supporting more photosynthetically active vegetation and hence having higher NDVI. PMID- 9737598 TI - Distribution of the Simulium damnosum complex on Bioko island, Equatorial Guinea, and the potential for onchocerciasis elimination by vector eradication. AB - Onchocerciasis is endemic on the island of Bioko, Equatorial Guinea, where it is transmitted by the 'Bioko form' of the Simulium damnosum complex, a cytospecies unique to the island. To determine the distribution of vector breeding, three dry season and two wet season expeditions were made in 1989, 1996 and 1997, and 226 of the island's 247 rivers (91.5%) were visited. Of these 226 rivers, 130 (58%) were flowing during the dry season, forty-five (20%) supported aquatic stages of Simuliidae of any species and twenty-five (11%) contained larvae or pupae of the S. damnosum complex. The twenty-one rivers not prospected were in the mountainous south of the island, where an additional seventeen rivers were reached but not satisfactorily prospected. Of these thirty-eight rivers, twenty-nine were considered highly likely to support vector breeding, bringing the total number of rivers which could harbour the vector during the dry season to fifty-four (21.9% of the island's total). Breeding was believed to be limited to river stretches below 1000 m altitude, and during the dry season the total length of those stretches which could support breeding on Bioko was estimated to be 1020 km. A combination of factors, including low river discharges during the dry season, the relatively low water temperature on Bioko, the suitability of limited stretches of most rivers as vector breeding sites and the close proximity of many rivers within a small geographical area, render the vector vulnerable to eradication by aerial treatment of rivers with insecticide. The isolation of the Bioko form of the S. damnosum complex suggests that reinvasion following treatment would be unlikely, and eradication of the vector might be achieved by a dry season larviciding programme in one or two years. PMID- 9737599 TI - Serum and skin surface antibodies and their associations with sheep biting lice, Bovicola ovis, on experimentally infested sheep. AB - The sheep biting louse (Bovicola ovis) feeds superficially on the skin of sheep but appears to stimulate an immune response. In this study we examined the association between louse infestation and serum and skin surface antibodies. Louse numbers were monitored on experimentally infested Polypay and Columbia ewes for two years and on their lambs in the second year. Serum and skin wash samples were tested for antibodies to soluble extracts of B. ovis, Stomoxys calcitrans and Musca autumnalis by enzyme linked immunosorbent assay (ELISA). In addition, the effects of skin wash extracts on B. ovis were examined in vitro. The titre of anti-B. ovis antibodies in the serum did not differ significantly between infested and naive ewes. However, there was an increase in serum antibody titre which coincided with periods of high louse density in ewes with high louse counts. Infested lambs had higher serum antibody levels than naive lambs. Substantial cross reactivity was evident among extracts of the different insects. Densities of lice on the ewes during population decline were negatively related to the titre of skin surface antibodies. Skin washings collected from sheep during B. ovis population decline reduced the number of louse progeny when incorporated into louse diet. These results indicate that B. ovis stimulates an immune response in sheep and suggest that compounds on the skin surface may play a role in the regulation of louse populations. PMID- 9737600 TI - Determination of dengue virus serotypes in individual Aedes aegypti mosquitoes in Colombia. AB - Adult Aedes aegypti mosquitoes were collected in Puerto Triunfo, central Colombia, where dengue is endemic, during a six month period. Viral infection within the head of each individual mosquito was identified by an immunofluorescent assay (IFA) using a flavivirus-specific monoclonal antibody. The dengue virus serotype, present in each flavivirus-positive specimen, was then determined in portions of the remaining thorax using IFAs with serotype-specific monoclonal antibodies. Among 2065 female Aedes aegypti collected and tested, twenty-four flavivirus-positive individuals were found (minimum infection rate 11.6%), three identified as dengue type-1 and twenty-one as dengue type-2 virus. This was consistent with the isolation of only these two serotypes of dengue virus from dengue fever patients within this town. No vertical transmission of dengue virus could be detected in 1552 male Aedes aegypti collected. This method is inexpensive, simple, rapid to perform and suitable for use in developing countries to identify and distinguish different serotypes of dengue virus in their vectors during eco-epidemiological investigations. PMID- 9737602 TI - Thanaka (Limonia acidissima) and deet (di-methyl benzamide) mixture as a mosquito repellent for use by Karen women. AB - The prevention and treatment of drug-resistant malaria is becoming increasingly difficult. On the Thai-Myanmar border multi-drug resistant strains of falciparum malaria are increasing and, because the malaria vector Anopheles bite outdoors during early evening, insecticide house-spraying or impregnated bednets provide only limited protection. Therefore, the protective efficacy of repellent formulations containing di-methyl benzamide (deet) and permethrin against local vectors was estimated, when applied to the skin, and their acceptability amongst pregnant Karen women who are at relatively high risk from malaria was assessed. Human landing catches of mosquitoes showed that almost complete protection was achieved using different formulations of 20% deet and 0.5% permethrin for up to 6 h. All-night collections from human subjects indicated that this repellent combination reduced exposure to malaria parasites by at least 65 and 85% for those transmitted by Anopheles minimus and An. maculatus, respectively, the two principal vectors in this area. Pregnant women in the camps preferred repellents which were mixed with 'thanaka', a root paste made from pulp of the wood apple tree, Limonia acidissima, used locally as a cosmetic. Apart from a temporary warming sensation where repellent thanaka was applied to the skin, the repellents were well tolerated. An intervention trial is currently in progress to determine whether deet mixed with thanaka can protect pregnant women against malaria in this part of the world. Bioassays using a laboratory strain of Aedes aegypti demonstrated that thanaka is itself slightly repellent at high dosages and the mixture with deet provides protection for over 10 h. This treatment would therefore also provide some personal protection against dengue, which is increasing locally, transmitted by Ae. aegypti and Ae. albopictus biting during the daytime. PMID- 9737601 TI - Comparative repellency of commercial formulations of deet, permethrin and citronellal against the mosquito Aedes aegypti, using a collagen membrane technique compared with human arm tests. AB - A collagen membrane technique, based on the membrane blood-feeding system of Cosgrove et al. (1994), was used to compare repellents against Aedes aegypti mosquitoes. Repellency was defined in terms of inhibition of probing (ED50 and ED90) after 5 min exposure. A direct comparison was made with repellency from probing after 5 min on five male volunteers. Four repellent products were compared with technical DEET as the standard. The liquid formulations tested were: Autan (20% deet; Bayer); Repel Plus (20% deet plus 0.05% permethrin; Boots); permethrin (Zeneca) and citronellal (Sigma). Membrane and arm tests gave similar results. Deet formulations required less active compound than citronellal for the same degree of repellency. Pure deet and Autan gave similar results, dose for dose. Permethrin was highly repellent at very low doses, but Repel Plus did not enhance the immediate repellency of deet. A technique using the same membrane system was developed to evaluate persistence of Autan, which declined to 75% after 1 h against Ae. aegypti, and to about 50% after 2-4 h. PMID- 9737603 TI - Genetics of refractoriness to Plasmodium falciparum in the mosquito Anopheles stephensi. AB - We previously selected a line of the malaria vector mosquito Anopheles stephensi refractory (resistant) to the human malaria parasite Plasmodium falciparum, using in vitro infections with P. falciparum gametocytes. This report presents data on the genetic background of refractoriness. The results of F1-crosses and backcrosses show that refractoriness to P. falciparum in our A. stephensi line is autosomal and semi-dominant to susceptibility. The expression of refractoriness is apparently affected by a cytoplasmic factor. Interpretation of data from the crosses by quantitative trait locus analysis shows that one gene or two unlinked interacting autosomal genes, or groups of closely linked genes, are involved. PMID- 9737604 TI - Dual infestation of a leopard by Wohlfahrtia magnifica and Lipoptena chalcomelaena. PMID- 9737605 TI - Incrimination of Lutzomyia cruzi as a vector of American visceral leishmaniasis. PMID- 9737606 TI - Biochemical monitoring of organophosphorus and carbamate insecticide resistance in Aedes aegypti mosquitoes from Trinidad. PMID- 9737607 TI - Effects of contrast on attraction of the housefly, Musca domestica, to visual targets. PMID- 9737608 TI - Malaria-infective biting at different hours of the night. PMID- 9737609 TI - Rearranging the deck chairs on the Titanic. PMID- 9737610 TI - Hume Memorial lecture. Prevention of spinal cord complications in aortic surgery. AB - Paraplegia or paraparesis after operations on the thoracic and abdominal aorta is a devastating event, both for the patient and the surgeon. While its incidence varies from under 1% with operations at the top and bottom of the aorta, its occurrence in the midportion of the aorta, just above the diaphragm, even in the best of hands exceeds 10%. Over a decade ago, Crawford et al (J Vasc Surg. 1986;3:389-404) introduced the use of inclusion and sequential clamping techniques for thoracoabdominal aneurysmectomy, lowering both morbidity and neurologic sequelae. Although these techniques have been widely adopted, newer ancillary adjuncts have been recommended by a number of investigators. This paper summarizes the possible causes of paraplegia secondary to the various operations on the aorta and analyzes the status and value of the various ancillary techniques in its prevention. PMID- 9737611 TI - Peter B. Samuels Award. Restenosis after percutaneous transluminal angioplasty. AB - BACKGROUND: Determine the feasibility of studying the natural history of the atherosclerotic plaque following percutaneous transluminal angioplasty (PTA), using duplex scanning. METHODS: Twenty-three patients with 40 stenoses (>70% and <5 cm in length) in the iliac and femoro-popliteal segments were studied by duplex scanning before PTA, on day 1, weekly for 8 weeks, and at 3 months, 6 months, and 1 year. The following measurements were made: thickness of the plaque, minimal lumen diameter (MLD), and peak systolic velocity ratio (PSVR). A PSVR >2.0 was used to indicate >50% lumen diameter reduction. RESULTS: Thirty stenoses were available for measurement and analysis. Mean reduction in plaque thickness after angioplasty was greater in echolucent plaques (2.33 +/- 0.9 mm) than echogenic plaques (0.83 +/- 0.6 mm; P < 0.0001). Successful angioplasty (PSVR <2.0) and increase in MLD in echolucent plaques was the result of plaque compression; in echogenic plaques, of wall dilatation. The incidence of restenosis (PSVR >2.0) at 6 months was 12 of 30 (40%) remaining unchanged at 1 year; of the lesions that restenosed, 33% recurred before week 8 and the remainder between weeks 8 and 24, suggesting different mechanisms. During follow up, all plaques showed "growth"; <2 mm in 17 (57%; group A) and >2 mm in the remaining 13 (43%; group B). The incidence of restenosis (PSVR >2.0) was 4 of 17 (23%) in group A and 8 of 13 (61%) in group B (P <0.05). CONCLUSION: Duplex scanning provides valuable information on both luminal diameter and plaque thickness; it may be used to study the natural history of plaques following angioplasty and also the effects of therapeutic agents aimed at reducing restenosis. PMID- 9737612 TI - A regional pedal ischemia scoring system for decision analysis in patients with heel ulceration. AB - PURPOSE: The objective of this study was to evaluate patients undergoing operative debridement for heel ulceration and to categorize pedal perfusion and its influence on therapeutic alternatives. METHODS: Patients with heel ulceration were stratified by arteriography and graded I (patent posterior tibial, PT), II (occluded PT/reconstituted from peroneal), III (PT reconstituted from dorsal pedal), IV (no PT reconstitution but visible heel tributaries), and V (avascular heel). RESULTS: From May 1992 through January 1997, 23 patients underwent operative treatment for 25 heel ulcers. The heel ischemia score stratified patients into two groups: 1, revascularization/debridement (71% grades I to III, 29% grade IV, 0% grade V); and 2, free tissue transfer with or without revascularization (100% grades IV, V). Cumulative functional limb salvage was 91% (BP), 60% (BP + TT), and 81% (TT) at 24 months (P = 0.15 log rank). CONCLUSION: The heel ischemia score may direct treatment of heel ulceration by identifying patients who will need vascularized tissue transfer early in their treatment regimen. PMID- 9737613 TI - Aorto-caval and ilio-iliac arteriovenous fistulae. AB - BACKGROUND: To determine optimal management of major abdominal arteriovenous fistulae and define factors affecting outcome. METHODS: We reviewed clinical data of 18 patients, 16 males and 2 females, who underwent repair of major abdominal arteriovenous fistulae between 1970 and 1997. RESULTS: Sixteen patients had primary fistula, caused by rupture of an atherosclerotic aortic or aortoiliac aneurysm into the inferior vena cava (IVC), iliac, or left renal vein. Two had secondary, iatrogenic arteriovenous fistulae. Seventeen patients (94%) were symptomatic, 11 (62%) had acute presentation. Fistula was diagnosed preoperatively in 8 (44%). Fistula closure (direct suture 16, patch 1, iliac vein ligation 1) was followed by aortoiliac reconstruction in all patients. Caval clip was placed in 3 patients. Early mortality was 6%; 7 patients had major complications. During follow-up (mean 6.1 years) 2 patients died of causes related to fistula closure. CONCLUSIONS: Rupture of aortoiliac aneurysms into the iliac veins or IVC carries a better prognosis than intraperitoneal, retroperitoneal, or enteric rupture. Although preoperative diagnosis is ideal, a high index of suspicion, careful repair avoiding pulmonary embolization, and blood salvage were all helpful in keeping morbidity and mortality low. Our data suggest that IVC interruption is seldom warranted. PMID- 9737614 TI - Percutaneous femoral puncture for endovascular treatment of occlusive arterial lesions. AB - BACKGROUND: Percutaneous femoral arterial access is a most important and difficult aspect of endovascular intervention, and the source of most complications. METHODS: A retrospective review was made of the authors' 9-year experience with 755 femoral punctures for the endovascular treatment of occlusive disease. The main focus was the evolving success rate with percutaneous arterial entry and the incidence of access-related complications. RESULTS: Cutdowns were frequent during the first 2 years, 54% and 17%, respectively, decreasing to 5% or lower by the third year. The incidence of femoral hematoma and other complications mirrored the same learning curve. After cutdown, wound infections and lymph leakage occurred in 2.4% each, and prolonged significant pain in 5%. CONCLUSIONS: Percutaneous puncture is a crucial skill in endovascular intervention. Practicing vascular surgeons can expect a significant learning curve. Performance can be optimized through intensive basic and advanced training and preceptorship. The cutdown approach is neither necessary nor acceptable for most endovascular procedures. PMID- 9737615 TI - Arterial thromboembolic events in patients with the factor V Leiden mutation. AB - BACKGROUND: The factor V Leiden mutation affects 6% of the United States population and is known to be associated with venous thrombosis. We identify, herein, 30 individuals with the Leiden mutation and known arterial thromboembolic events. METHODS: The factor V mutation was assessed using polymerase chain reaction. RESULTS: In the 16 patients sustaining a cerebrovascular accident, the mean age was 44.1 and 11 (69%) were younger than 50. Similarly, the 13 patients presenting with an acute myocardial infarction were relatively young with a mean age of 45.5, and 9 (65%) patients presented at less than 50 years of age. Radiographic information was available for 19 patients in this study. No significant arterial atherosclerotic disease was demonstrated in 18 (95%) of these patients. CONCLUSIONS: This study demonstrates an association between the factor V Leiden mutation and the development of unexplained arterial thromboembolic events, especially in younger patients without existing atherosclerotic disease. PMID- 9737616 TI - Revision of failing lower extremity bypass grafts. AB - BACKGROUND: Color-duplex ultrasound (CDU) surveillance of arterial bypass grafts has been validated, but the natural history of "failing" grafts remains poorly defined. Our purpose was to compare failing grafts having prophylactic revision with those that did not. METHODS: Postoperative duplex surveillance was performed in an accredited vascular laboratory for all lower extremity bypass grafts performed at a single institution. Eighty-five infrainguinal grafts (57 vein, 21 polytetrafluoroethylene (PTFE), and 7 composite grafts) in 83 patients were identified as failing by accepted criteria. Twenty-five grafts were revised early (early), 20 grafts revised more than 2 months after the initial CDU-abnormality (late), and 40 grafts were not prophylactically revised (no revision) at any time. RESULTS: The three groups were not different (P > 0.10) with regard to gender, age, level of bypass, type of conduit, location of stenoses, or timing of abnormality after surgery. No revision patients more frequently had diffuse low peak systolic flow velocity (PSV) as the CDU abnormality (P = 0.013). Cumulative primary patency was significantly better at 12 months (P = 0.028) in the no revision group (78.9%) compared with early grafts (43.1%) or late grafts (63.8%), and this difference remained significant when low PSV grafts were excluded from analysis. However, assisted primary patency, secondary patency, and limb salvage rates did not differ between the three groups (P > 0.10). CONCLUSIONS: Our experience in this retrospective study contradicts other reports supporting the efficacy of prophylactic graft revision for grafts identified as failing by currently accepted CDU criteria. Refinement of CDU criteria to more accurately predict graft thrombosis is needed. PMID- 9737617 TI - Expanded indications for the treatment of postcatheterization femoral pseudoaneurysms with ultrasound-guided compression. AB - BACKGROUND: The purpose of this study was to define the factors that predict successful ultrasound-guided compression repair (UGCR) of postcatheterization femoral pseudoaneurysms (PA) and to determine risks for recurrence, the most appropriate follow-up, and the optimal management of compression failures and recurrences. METHODS: A retrospective chart review was made. RESULTS: UGCR thrombosed 52 of 60 PA (87%). Predictors of compression failure were PA size of 8 cm and an associated arteriovenous fistula (AVF). AVF was the only predictor of recurrence. All seven recurrences (13%) were discovered on the first follow-up scan. Four were thrombosed with additional UGCR. Late rescanning after a mean of 264 days identified no recurrences. Four anticoagulated patients failed initial UGCR but were thrombosed in another session when their anticoagulation was briefly reversed. CONCLUSIONS: UGCR should be the initial management of PA because it is safe, effective, and durable. Temporary discontinuation of anticoagulation and multiple prolonged compression sessions may help treat recalcitrant cases. One follow-up scan is adequate for most patients. Recurrences should be initially treated with repeat UGCR. PMID- 9737618 TI - Malignant renal tumor with extension to the inferior vena cava. AB - BACKGROUND: Management of malignant renal tumors involving the inferior vena cava (IVC) depends on tumor extension within the cava. METHODS: Of 295 patients treated for renal cancer, propagation of tumor mass through the renal vein to IVC was seen in 22 (7%) patients. Cephalad extension of the tumor was suprarenal: infrahepatic in 12, retrohepatic in 6, and within the right atrium in 4 patients. All patients had radical nephrectomy, cavotomy, and complete resection of tumors except 1 with diffuse peritoneal metastasis. RESULTS: Twenty-one patients had curative resections. No operative deaths and no instances of pulmonary embolism or exsanguination occurred. Seventeen patients were alive at 2 years and 12 at 5 years, resulting in 77% and 55% survival rates, respectively. CONCLUSIONS: An aggressive approach for vena cava involvement from malignant renal neoplasms resulted in prevention of tumor embolus, minimization of blood loss, and maintenance of venous return to the heart. PMID- 9737619 TI - Compensatory arterial enlargement is a common pathobiologic response in early atherosclerosis. AB - BACKGROUND: Human arteries are dynamic conduits that respond to different stimuli by remodeling their structure and size. Arterial dilatation has been shown to occur in moderate and advanced atherosclerosis in studies that evaluated only one artery, either coronary, carotid, or superficial femoral artery (SFA). The purpose of this study was to quantify and compare compensatory arterial enlargement throughout the peripheral vascular system in early atherosclerosis. METHODS: Seventy-two patients (40 male, 32 female, mean age 67 +/- 12 years) underwent transcutaneous B-mode ultrasound imaging during routine examinations. Thirty-nine carotid, 19 aorta, 19 iliac, 23 common femoral (CFA), 21 SFA, and 23 popliteal arteries were longitudinally imaged. Eight healthy volunteers (6 male, 2 female, mean age 27 +/- 2.2 years) had the same arteries evaluated (n = 48). Internal diameter (ID) and external diameter (ED) were measured in disease-free areas and in paired adjacent areas exhibiting increased intima-media thickening (IMT) and small atherosclerotic plaques. The percent change in ID, ED, IMT, and plaque thickness were calculated. RESULTS: There was no observed change in ID or ED in all arteries of the healthy volunteers. When compared with normal vessel segments, all arteries demonstrated a marked decrease in ID and increase in ED in areas of small, hemodynamically insignificant plaque. The aorta had a 6.00% +/- 1.92% increase in ED, which was significantly less than the percent increase in ED observed in carotid (8.14 +/- 4.5%. P = 0.05), CFA (9.73 +/- 3.54%, P = 0.0001), SFA (9.15 +/- 4.25%, P = 0.005), and popliteal arteries (9.67 +/- 4.34, P = 0.002). In all arteries there was a strong correlation between plaque thickness and percent change in ED with the best correlation observed in the popliteal artery (R2 = 0.823, P < 0.0001). IMT was significantly increased in all normal vessel segments of the patients when compared with the healthy volunteers (P < 0.001). CONCLUSION: All peripheral arteries dilate in response to intima media thickening and early atherosclerotic plaque formation. This adaptive response occurs at the site of the lesion to preserve luminal area. The percent change in ED is strongly related to plaque thickness and is greatest in the more distal arteries. PMID- 9737621 TI - Nontraumatic lower-extremity acute arterial ischemia. AB - BACKGROUND: The outcome of arterial bypass reconstruction in the setting of acute arterial ischemia has not been well defined. METHODS: This retrospective review consists of 71 consecutive patients (54 with native arterial thrombosis, 17 with graft thrombosis) who underwent an urgent/emergent arterial bypass reconstruction for acute arterial ischemia with threatened limb viability. RESULTS: The 30-day mortality and major amputation rates were 9.9% and 7.1%, respectively. Death, limb loss, or both, were associated with a paralytic limb (P = 0.001) and congestive heart failure (P = 0.03). Overall, 45 of 71 (63%) patients were discharged with limb salvage and ambulatory function. Cumulative graft patency was 77% and 65% at 1 and 2 years, respectively, and closely approximated the 1- and 2-year limb-salvage rates of 76% and 63%, respectively. CONCLUSIONS: Arterial bypass reconstructions appear warranted in acute arterial ischemia, in that a majority of patients retain a functional viable limb. Late graft thrombotic complications limit long-term benefit. PMID- 9737620 TI - Combined coronary artery bypass grafting and abdominal aortic aneurysm repair. AB - BACKGROUND: We report here the results of combined coronary artery bypass grafting (CABG) and abdominal aortic aneurysm (AAA) repair and the factors associated with higher mortality following this procedure. METHODS: The authors performed a retrospective chart review of 26 patients who underwent combined CABG and AAA repair between March 1990 and October 1996. RESULTS: No postoperative myocardial infarction or major cardiac complications were noted. A morbidity rate of 38% (n = 10) and mortality rate of 11% (n = 3) were noted. Comparative analysis of nonsurvivors (n = 3) versus survivors (n = 23) revealed the following: ejection fraction (EF) was significantly lower (33% +/- 3% versus 44% +/- 14%, P < 0.05), duration of cardiopulmonary bypass (CPB) was significantly longer (239 +/- 122 minutes versus 141 +/- 54 minutes, P < 0.05), and incidence of postoperative respiratory failure (67% versus 17%, P = 0.001) were significantly higher in nonsurvivors. No differences in mean age, gender distribution, incidence of hypertension or diabetes were noted between the groups. CONCLUSIONS: Combined CABG and AAA repair protected patients from postoperative aneurysm rupture and myocardial infarction. Poor EF, prolonged CPB, and postoperative respiratory failure were associated with higher mortality. PMID- 9737622 TI - An association between periodontal disease and peripheral vascular disease. AB - BACKGROUND: Periodontal disease has been shown to be associated with increased risk of coronary heart disease. Because coronary heart disease and peripheral vascular disease (PVD) have similar pathophysiologies, we hypothesized that periodontal disease might be a risk factor for PVD. METHODS: Using the combined data from the Normative Aging Study and Dental Longitudinal Study of the US Department of Veterans Affairs, we examined the relationship between PVD and periodontal disease. Multivariate logistic regression analysis was used. RESULTS: Over the 25 to 30 years of follow-up, 80 of these initially healthy subjects developed PVD. Compared with controls (n = 1,030), subjects with clinically significant periodontal disease at baseline had a 2.27 increment in the risk of developing PVD (95% confidence interval 1.32 to 3.9, P value = 0.003). CONCLUSIONS: Periodontal disease emerged as a significant independent risk factor for PVD in a multivariate analysis that adjusted for other established risk factors. PMID- 9737623 TI - Etiology of peripheral arterial thromboembolism in young patients. AB - BACKGROUND: No prior studies have explored the etiology of peripheral arterial thromboembolic events (PATE) in younger patients. Therefore, we analyzed the sources of these events in patients <50 years of age over a recent 10-year period. Diagnostic and work-up strategies will be proposed based on the presence of cardiac or atherosclerotic risk factors. PATIENTS AND METHODS: The sources of emboli were classified as (1) conventional (cardiac or arterioarterial), (2) unconventional, or (3) unknown. A statistical analysis of risk factors that, if absent, would suggest an unconventional cause was performed. Risk factors included those for cardiac and atherosclerotic disease: coronary artery disease (CAD), valvular disease, smoking, arrhythmia, hypertension, or diabetes mellitus. RESULTS: Overall, 51 patients were identified. Twenty-nine patients (57%) had unconventional causes (8 paradoxical emboli, 4 possible paradoxical emboli, 12 hypercoagulable states, 3 white clot syndromes, and 2 cervical ribs), 17 (33%) had conventional causes, and 5 (10%) were unknown. When the number of cardiac risk factors was < or =1, excluding smoking, the probability of a conventional source was zero, in contrast to 100% if the number of risk factors was >1. When the following risk factors were absent, there were significantly more unconventional than conventional sources of emboli (P < 0.001): smoking (100% versus 0%), CAD (93% versus 7%), arrhythmias (83% versus 17%), hypertension (93% versus 7%), and diabetes mellitus (81% versus 19%). Patients with a conventional source were significantly older (44 versus 38 years). CONCLUSIONS: The "unconventional" causes of PATE were responsible for a higher percentage of cases in young patients. An analysis of the number of risk factors was useful in predicting which patients suffered embolic events from conventional sources, with the critical number being >1 (excluding smoking). Therefore, when younger patients present with PATE, and are found to have < or =1 identifiable cardiac risk factor, their work-up should be directed toward the unconventional sources first. PMID- 9737624 TI - Factors affecting clinical outcome following endoscopic perforator vein ablation. AB - BACKGROUND: Despite good outcomes reported with minimally invasive, subfascial endoscopic perforator surgery (SEPS), some patients demonstrate poor healing or recurrence of venous ulcers. The goal of this study was to identify factors that lead to failure of SEPS. METHODS: Forty-eight consecutive patients who had undergone 57 SEPS procedures were analyzed. Mean follow-up was 17 +/- 2 months (range 2 weeks to 52 months). RESULTS: All active ulcers (n = 22) at the time of surgery healed in an average of 99 +/- 37 days (range 11 to 670). Eight limbs had poor healing of their ulcer (>40 days); five (9%) new/recurrent ulcers developed postoperatively. Deep venous obstruction was associated with delayed ulcer healing (316 +/- 171 versus 51 +/- 14 days, P < 0.01) and ulcer recurrence (P < 0.0001). Poor ulcer healing and recurrence were not associated with lipodermatosclerosis, edema, ulcer duration >3 months, or previous recurrences. Ulcer size >2 cm (P < 0.05) and combined ilio-femoral and popliteal/tibial reflux were associated with poor ulcer healing (P < 0.05). CONCLUSIONS: SEPS could not prevent recurrent or new ulceration in 9% of limbs. Venous outflow obstruction was associated with ulcer recurrence and prolonged ulcer healing. Multilevel deep venous reflux and ulcer size >2 cm were also associated with delayed healing. PMID- 9737625 TI - Is thigh saphenectomy a necessary adjunct to high ligation and stab avulsion phlebectomy? AB - BACKGROUND: Controversy still exists as to whether the thigh saphenous vein should be stripped concomitant with high ligation and phlebectomy. METHOD: A total of 218 procedures were retrospectively grouped into three groups: group 1, 10 limbs with visible, duplex scan-confirmed varicose veins of the thigh saphenous vein; group 2, 13 saphenous veins with varices that were not clinically evident; group 3, 195 limbs with incompetent saphenous veins without thigh saphenous varices. RESULTS: Five limbs in group 1 were treated by high ligation, phlebectomy, and thigh saphenectomy. All did well. Five had high ligation and phlebectomy only. Two developed painful phlebitis, and two had residual varices in the saphenous vein. Group 2 and group 3 were treated by high ligation and phlebectomy. One group 2 limb developed saphenous phlebitis. Five limbs in group 3 developed recurrent veins that were removed in the office. CONCLUSION: Thigh saphenectomy is only required when there are visible, duplex scan-confirmed varices of the thigh saphenous itself, or when the procedure is performed for severely symptomatic patients or those with advanced stasis changes. PMID- 9737626 TI - Clinical benchmark for healing of chronic venous ulcers. Venous Ulcer Study Collaborators. AB - BACKGROUND: To determine the results of standardized ulcer treatment regimes and effects of the oral thromboxane A2 antagonist Ifetroban (250 mg daily) on healing of chronic lower-extremity venous stasis ulcers. METHODS: In a prospective, randomized, double blind, placebo-controlled multicenter study, 165 patients were randomized to Ifetroban (n = 83) versus placebo (n = 82) for a period of 12 weeks. Both groups were treated with sustained graduated compression and hydrocolloid. Ulcer size was measured weekly by tracings and computerized planimetry. A total of 150 patients completed the study. RESULTS: Complete ulcer healing was achieved after 12 weeks in 55% of patients receiving Ifetroban and in 54% of those taking a placebo with no significant differences; 84% of ulcers in both groups achieved greater than 50% area reduction in size. CONCLUSIONS: These results are likely to be useful as a benchmark for comparison with other treatment protocols concerning the care of chronic lower-extremity stasis ulcers. PMID- 9737628 TI - New, angle-independent, low cost Doppler system to measure blood flow. AB - BACKGROUND: A new Doppler flowmeter using a special transducer forming two ultrasound beams to insonate vessels was developed. This low-cost flowmeter allows the flow to be measured independently of the angle of insonation. METHODS: Sixty-five flow determinations were made in the carotid arteries of five pigs. The flowmeter measured the flow in an artery bled into a calibrated vessel. Results were compared with the true volume of the blood captured. RESULTS: On average, 13 measurements per animal were made at different flow rates (80 to 600 mL/min). The flowmeter measured flow-volume rates in mL/min were found to be within +/- 15% of the cylinder captured volume. CONCLUSIONS: The accuracy of the angle-independent flowmeter is comparable with that of flowmeters currently used clinically. The small size and portability, low operator dependency, and low cost suggest potential for continuous acute and chronic-flow monitoring and a potential for an implantable flow-monitoring device. PMID- 9737627 TI - Orally administered heparin for preventing deep venous thrombosis. AB - BACKGROUND: Sodium N-(8-[2-hydroxybenzoyl] amino) caprylate (SNAC) is an acetylated amino acid molecule that facilitates the gastrointestinal absorption of heparin. This study was undertaken to evaluate the efficacy of orally administered combination SNAC:heparin in preventing deep venous thrombosis in a standard rat model. METHODS: Forty-four adult male Sprague-Dawley rats were randomly divided into five groups: group I control, group II SNAC, group III oral heparin, group IV combination SNAC:heparin, and group V intravenous heparin. Thirty minutes after drug administration, the internal jugular vein was bathed in a sclerosant mixture for 2 minutes and reexplored at 120 minutes. Activated partial thromboplastin times (aPTT) were measured in 30 rats equally divided into three groups: group I SNAC, group II oral heparin, and group III combination SNAC:heparin. Forty-five minutes posttreatment, blood was obtained for aPTT levels. RESULTS: The incidence of deep venous thrombosis in the control group was 89% (8 of 9) versus 25% (2 of 8) in the combination SNAC:heparin group (p < 0.01). There was also a significant reduction in clot weight among groups. Combination SNAC:heparin significantly increased aPTT levels compared with SNAC or oral heparin alone. CONCLUSION: In a rat model of venous thrombosis, combination of orally administered heparin:SNAC elevated aPTT levels and significantly reduced the formation of deep venous thrombosis. PMID- 9737629 TI - Three-dimensional vascular imaging using Doppler ultrasound. AB - BACKGROUND: We have evaluated the efficacy of using three-dimensional reconstruction of amplitude Doppler imaging data to quantitatively assess carotid artery bifurcation stenoses. METHODS: Sixty-four consecutive frames of amplitude (power) Doppler images are stored to be reassembled into a three-dimensional image representing the patent lumen. These images can then be rotated by any angle necessary to clearly view the vascular anatomy and to make quantitative ultrasound caliper measurements of the stenotic lumen and normal vessel caliber. RESULTS: Three-dimensional Doppler images accurately classified 53 of 61 vessels (87%) into categories of stenosis compared with angiography. All stenoses with >60% diameter reduction were detected and classified as such, for a sensitivity of 100%. CONCLUSIONS: Three-dimensional vascular imaging based on amplitude (power) Doppler data provides an accurate noninvasive technique for quantitative diagnosis of carotid bifurcation atherosclerotic disease, with selectable viewing projections that eliminate vessel overlap and other artifacts, and complements the hemodynamic data already available with two-dimensional duplex ultrasound. PMID- 9737630 TI - Cost management strategies for carotid endarterectomy. AB - BACKGROUND: We developed a model for capitation and global pricing for carotid endarterectomy. METHODS: A care algorithm for diagnosis, perioperative management, and postoperative care using cost data was developed. Perioperative care charges were extrapolated from a 1-year experience and applied to models to determine pricing for a 1-year global fee and a 5-year capitated contract. RESULTS: Global pricing was estimated at $12,071 per patient while a capitated price for 5-year care was $17,175. Based on the age mix of the population, a per member, per month cost could be calculated assuming a frequency of 414 procedures per 100,000 patients over age 65 and 31 procedures per 100,000 patients under 65. Sources of costs were extensive preoperative diagnostic testing, particularly angiography, brain imaging, and cardiac evaluation. CONCLUSIONS: Global pricing and capitation are both feasible for carotid endarterectomy. Each approach has unique risks and benefits. PMID- 9737631 TI - Acute occlusion of the abdominal aorta. AB - BACKGROUND: Acute aortic occlusion most commonly results from aortic saddle embolus or thrombosis of an atherosclerotic abdominal aorta. The purpose of this study was to review the experience at a university hospital to better define the diagnosis and management of this uncommon process. METHODS: A retrospective chart review was performed from patients admitted to Emory University Hospital with acute occlusion of the abdominal aorta from 1985 through 1997. RESULTS: Thirty three patients were identified. In group EMB (n = 16), occlusion was due to saddle embolus. In group IST (n = 17), occlusion was attributed to in situ thrombosis of a severely diseased aorta. Operative procedures performed included transfemoral embolectomy (15), aorto-bifemoral bypass (9), axillobifemoral bypass (5), fasciotomy (3), and thrombolysis (1). The in-hospital mortality rate was 21% (31% EMB, 12% IST), and morbidity was significant and included mesenteric ischemia (6%), bleeding complications (9%), subsequent amputation (12%), renal failure (15%), recurrent embolization or thrombosis (21%), and cardiac complications (42%). CONCLUSIONS: Acute aortic occlusion has tremendous morbidity and mortality even with optimal surgical care. PMID- 9737632 TI - The experience of an academic medical center with endovascular treatment of abdominal aortic aneurysms. AB - BACKGROUND: Endovascular repair of abdominal aortic aneurysms (AAA) is gaining momentum although it is not yet approved in the United States by regulatory agencies. The Endovascular Grafting System (EGS), the first device to enter clinical trials in 1993, is now in phase III testing. METHODS: We reviewed the first 50 patients to undergo an EGS repair of AAA over 24 months at our institution. Results were compared with 69 patients who underwent open repair during the same time period by the same surgeon. RESULTS: Devices were successfully implanted in 47 of 50 (94%) patients. Three were converted to standard repair. Although length of stay was shorter, costs were similar. Follow up was 3 to 24 months. Perigraft flow was noted in 33% at discharge; 73% of those stopped either spontaneously or with coiling. Three graft limbs occluded, requiring thrombolytic therapy. CONCLUSIONS: The EGS repair of AAA is feasible and effective. Cooperation between surgery and radiology is important for the success of a new endovascular program. PMID- 9737633 TI - Lower extremity vascular reconstruction and endovascular surgery without preoperative angiography. AB - BACKGROUND: Recent studies have shown the feasibility of performing lower extremity revascularization based on noninvasive vascular studies alone. METHODS: We undertook a prospective study of patients with lower extremity ischemia who underwent revascularization without preoperative angiography. Preoperative evaluation was done with noninvasive studies including segmental pressures, ankle arm index, duplex scan, and selective use of magnetic resonance angiography. Intraoperative angiography and intra-arterial pressure measurements were used prior to revascularization. Standard patency analysis and follow-up examination were performed. RESULTS: In all, 47 patients underwent 65 procedures (27 iliac, 38 infrainguinal) over a 3-year period. Intraoperative angiography and operative findings correlated with the noninvasive studies. There was one immediate failure, and life table analysis demonstrated primary patency rates of 92% for iliac reconstruction (29 months) and 82% for infrainguinal reconstruction (40 months). CONCLUSION: Preoperative evaluation for lower extremity revascularization utilizing only noninvasive vascular testing gives satisfactory results and is a safe and potentially durable alternative to routine preoperative angiography in most cases. PMID- 9737634 TI - Peripherally inserted central catheters revisited. AB - BACKGROUND: This study compares central venous catheters (CVC) and peripherally inserted central catheters (PICC) for indications for insertion, complications, and economic impact. METHODS: A retrospective review of 838 (283 CVC, 555 PICC) consecutively placed venous catheters reflected 49,365 CVC and 11,814 PICC days. RESULTS: There were 57 (20%) complications in the CVC group, 197 (35%) complications in the PICC group. PICC were associated with a statistically significant increase in the incidence of catheter malfunction (P = 0.0005), arm vein phlebitis (P = 0.0004), and overall complications (P = 0.00001). A higher complication rate was noted in PICC inserted for chemotherapy (P = 0.00001) and parenteral hyperalimentation administration (P = 0.04). Charges for inpatient insertion of PICC and CVC were $500 and $2,500, respectively. CONCLUSIONS: PICC have a significantly higher complication rate than CVC. PICC provide cost effective central access of 2 to 3 weeks' duration, reserving operatively placed CVC for longer access requirements. PMID- 9737635 TI - Cardiac tamponade from central venous catheters. AB - BACKGROUND: This retrospective study was undertaken to determine the mechanism by which cardiac tamponade (CT) occurs after placement of central venous catheters (CVC), and to determine if physicians are aware of this potentially lethal complication. MATERIALS AND METHODS: Twenty-five previously unreported cases of CT from CVC were reviewed. The chest radiographs and postmortem records were reviewed when available. Two hundred physicians were interviewed about their knowledge of CT from CVC. They were specifically asked if they had reviewed the three-volume video, "CVC Complications," that was sent by the Food and Drug Administration to all hospitals where CVC are inserted. RESULTS: All postinsertion chest radiographs showed the tip of the catheter to be within the pericardial silhouette. All patients developed unexplained hypotension from hours to 1 week after CVC placement. Eight patients complained of chest tightness, 12 of shortness of breath, and 15 were noted to have air hunger. The electrocardiogram showed inferior wall injury in 7 patients. None of the physicians surveyed had seen the FDA video. CONCLUSIONS: Cardiac tamponade from central venous catheters is preventable if the tip of the catheter is outside the cardiac silhouette on chest radiograph. Any patient with a CVC in place who develops unexplained hypotension, chest tightness, or shortness of breath should have an emergency echocardiogram to rule out cardiac tamponade. PMID- 9737636 TI - A comparison of surgery for neurogenic thoracic outlet syndrome between laborers and nonlaborers. AB - OBJECTIVE: To determine factors of outcome following surgical intervention for neurologic thoracic outlet syndrome (NTOS). METHODS: In a retrospective study of patients surgically treated for NTOS, outcome was evaluated by postoperative symptoms and the ability of patients to return to work. RESULTS: Good, fair, and poor results were obtained in 26 (48%), 21 (39%), and 7 (13%) patients, respectively. The best predictor of a good outcome was occupation. Nonlaborers were more likely to have good outcome (21 of 32, 66%) when compared with laborers (5 of 22, 23%; P = 0.0025). Only 6 of 20 (30%) laborers were able to return to their original occupation compared with 17 of 26 (65%) nonlaborers (P = 0.036). CONCLUSIONS: Laborers with NTOS are less likely to have a good result from surgical intervention, are unlikely to return to their original occupation, and may require retraining for a non-labor-intensive occupation if they cannot return to their original work. PMID- 9737637 TI - Transposed basilic vein versus polytetrafluorethylene for brachial-axillary arteriovenous fistulas. AB - BACKGROUND: Both transposed basilic vein (BV) and polytetrafluorethylene (PTFE) upper arm arteriovenous fistulas (AVF) are common angioaccess operations. To evaluate the patency and complication rates after AVF, a concurrent series of patients was reviewed. METHODS: Ninety-eight patients underwent brachial artery to axillary vein AVF: 30 BV and 68 PTFE. The PTFE grafts were performed in the standard fashion, whereas the basilic veins were translocated subcutaneously to the brachial artery. RESULTS: Risk factors were similar between the two groups. Basilic vein AVF had better patency at 24 months (70% BV versus 46% PTFE, P = 0.023). The dialysis access complications were higher in the BV group (20%) versus PTFE (5%), but the PTFE group had a higher infection rate (10%) than BV (0%). CONCLUSIONS: The primary and secondary patency rates were superior in the BV AVFs. The BV AVF preserves the venous outflow tract after AVF thrombosis for a future PTFE AVF operation. PMID- 9737638 TI - An aggressive local approach to vascular graft infection. AB - PURPOSE: To present the use of sartorius myoplasty (SM) and superficial femoral vein (SFV) in a graft-sparing approach to vascular graft infection. METHODS: Twenty-five patients were treated for Szilagyi grade III groin infections during the last 10 years. Fifteen presented early (E), mean 2 months; 10 late (L), mean 6.5 years. Bacteria E/L: Staphylococcus epidermidis 1/7, S aureus 6/0, other gram positive 1/0, gram negative 4/1, mixed 6/0 (one pseudomonas). There were 13 aortofemoral (AF), 5 crossover, and 7 femoral distal reconstructions. SM was used to cover exposed grafts after radical debridement. When the graft was free floating or bleeding, this segment was removed and replaced with SFV. The remaining infection was controlled with antibiotics. RESULTS: One of 10 patients treated by SM alone required SFV replacement for bleeding. Four of 9 AFs treated by partial SFV replacement +/- SM had persistent infection treated by complete graft removal in 3. Six grafts were removed electively in lower risk situations. There was no perioperative mortality, and no early or late limb loss. CONCLUSIONS: This experience supports an attempt at aggressive local treatment in frail patients. PMID- 9737639 TI - Improved selection criteria for ordering stat venous ultrasounds from the emergency department. AB - BACKGROUND: The accuracy and convenience of venous ultrasound (VU) to exclude deep vein thrombosis (DVT) has led to indiscriminate use and low positive yield rates. METHODS: A total of 256 patients were referred from our emergency department (ED) for stat VU during a 2-year period (1995 to 1996). The VUs were interpreted as normal in 198 (77%). Positive findings were discovered in 58 (23%), with DVT accounting for 43 (17%). Retrospective multivariant analysis was used to identify predictive indicators. RESULTS: Unilateral leg swelling/edema identified 36 of 40 (90%) patients with DVT and 8 of 10 (80%) with other thrombotic disorders (saphenous and/or chronic venous thrombosis). A history of leg pain with prior DVT or recent trauma < or =3 days' duration increased DVT duration to 98% (39 of 40). Using these criteria, a 47% charge reduction would have been recognized. CONCLUSIONS: Improving ED screening criteria can safely increase yield rate and reduce charges with minimal loss of VU sensitivity. PMID- 9737640 TI - Current and future treatment of hyperlipidemia: the role of statins. AB - Hyperlipidemia is recognized as one of the major risk factors for the development of coronary artery disease and progression of atherosclerotic lesions. Dietary therapy together with hypolipidemic drugs are central to the management of hyperlipidemia, which aims to prevent atherosclerotic plaque progression, induce regression, and so decrease the risk of acute coronary events in patients with pre-existing coronary or peripheral vascular disease. In patients at high risk of coronary artery disease but without evidence of atherosclerosis, treatment is designed to prevent the premature development of coronary artery disease, whereas in those with hypertriglyceridemia, treatment aims to prevent the development of hepatomegaly, splenomegaly, and pancreatitis. The 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors, or statins, are the most potent lipid lowering agents currently available, and their use in the treatment of hyperlipidemia provides the focus for this review. Particular emphasis is given to cerivastatin, a new HMG-CoA reductase inhibitor that combines potent cholesterol-lowering properties with significant triglyceride-reducing effects. Recently completed primary and secondary intervention trials have shown that the significant reductions in low-density lipoprotein (LDL) cholesterol achieved with statins result in significant reductions in morbidity and mortality associated with coronary artery disease as well as reductions in the incidence of stroke and total mortality. Such benefits occur early in the course of statin therapy and have led to suggestions that these drugs may possess antiatherogenic effects over and above their capacity to lower atherogenic lipids and lipoproteins. Experimental studies have also shown statin-induced improvements in endothelial function, decreased platelet thrombus formation, improvements in fibrinolytic activity, and reductions in the frequency of transient myocardial ischemia. PMID- 9737641 TI - Preclinical safety evaluation of cerivastatin, a novel HMG-CoA reductase inhibitor. AB - Cerivastatin is a new but structurally distinct 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor ("statin"). It effectively decreases low density lipoprotein (LDL) cholesterol at 1% of the doses of other currently available statins. The toxicology of cerivastatin was evaluated in a comprehensive program of studies including: (1) single- and multiple-dose toxicity studies in rats, mice, minipigs, dogs, and monkeys; (2) reproductive toxicity studies in rats and rabbits; (3) in vitro and in vivo mutagenicity assays in rats and mice; and (4) carcinogenicity studies in rats and mice. In addition, studies were undertaken to investigate the effects of cerivastatin on lens opacity, testicular tissue, and hemorrhage in dogs. Oral administration of single and multiple doses of cerivastatin over periods ranging from 4 weeks to 24 months was generally well tolerated. Adverse effects were similar to those observed with other statins and primarily involved the liver and muscle tissue. At the high doses used in the toxicologic studies, cerivastatin caused elevations in serum transaminases and creatine phosphokinase levels as well as some degeneration of muscle fibers in rats, mice, dogs, and minipigs. In dogs, the species most sensitive to statins, cerivastatin caused erosions and hemorrhages in the gastrointestinal tract, bleeding in the brain stem with fibroid degeneration of vessel walls in the choroid plexus, and lens opacity. Apart from minor morphologic changes in the testicular tissue of dogs--the only organ for which a comparably low margin of safety was observed--cerivastatin had no significant effects on the male or female reproductive system. Cerivastatin also caused no primary embryotoxic or teratogenic effects. With the exception of cerivastatin-induced effects on the eyes and testicles, administration of mevalonic acid reversed the toxicologic effects of cerivastatin, indicating that the toxic effects were related to its mode of action and not to any intrinsic toxicity of the molecule itself. There was no evidence that cerivastatin had any mutagenic effects and, in contrast to other statins, high doses of cerivastatin did not induce tumors in rats. The main metabolite of cerivastatin was well tolerated systemically in all animals, including dogs. Overall, cerivastatin has a similar toxicologic profile to other statins and is a well-tolerated HMG-CoA reductase inhibitor. PMID- 9737642 TI - Preclinical and clinical pharmacology of cerivastatin. AB - Cerivastatin, a new, entirely synthetic, and enantiomerically pure 3-hydroxy-3 methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor, is pharmacologically potent and hepatically selective, with an uncomplicated pharmacokinetic profile. In vitro and acute in vivo studies in animals demonstrated that cerivastatin is markedly more pharmacologically potent than other statins. In rats and dogs, cerivastatin inhibited hepatic cholesterol synthesis at concentrations 100-150 times lower than lovastatin. Cerivastatin's potent inhibition of HMG-CoA reductase (the rate-limiting step in cholesterol biosynthesis) is confirmed by its cholesterol-lowering properties, combined with significant triglyceride decreasing effects, and dose-dependent increases in low-density lipoprotein (LDL) receptor binding in the liver. The antiatherogenic effects of cerivastatin extend beyond serum lipid and lipoprotein reductions to potent inhibition of migration of smooth muscle cells in vitro and reductions in the accumulation of cholesterol ester in the arterial tissue of rabbits. The high pharmacologic potency of cerivastatin, coupled with high liver selectivity, enable cerivastatin to be administered at 1-5% of the dose of currently available HMG-CoA reductase inhibitors. At ultra-low doses in the range 0.01-0.8 mg/day, cerivastatin proved to be both safe and well tolerated when administered to healthy volunteers in a series of ascending single- and multiple-dose studies. Cerivastatin has an uncomplicated pharmacokinetic profile; it can be administered to both young and elderly patients, male and female, without the need for dosage adjustments. Because no clinically significant pharmacokinetic drug interactions occur with cerivastatin, it may be the preferred HMG-CoA reductase inhibitor for patients on multiple-drug therapy including warfarin and digoxin. PMID- 9737643 TI - Clinical efficacy of cerivastatin: phase IIa dose-ranging and dose-scheduling studies. AB - Phase IIa clinical studies with cerivastatin--including 2 pilot US and European dose-ranging studies, and 1 US dose-scheduling study--were conducted to establish a dosage regimen and effective therapeutic doses of cerivastatin in the treatment of hypercholesterolemia. Both dose-ranging studies included a 10-week dietary run in, to which placebo was added in the last 6 weeks, before patients (n = 385) were randomized to 1 of 6 4-week treatment groups: cerivastatin (0.025, 0.05, 0.1, and 0.2 mg/day), 40 mg/day lovastatin (US), 20 mg/day simvastatin (Europe), or placebo. The dose-scheduling study also included a 10-week dietary run-in and 6-week single-blind placebo run-in phase, before patients (n = 319) were randomized to 4 weeks of treatment with either 0.1 mg cerivastatin twice daily, 0.2 mg cerivastatin with the evening meal, 0.2 mg cerivastatin at bedtime, or placebo in a 2:2:2:1 ratio. The 4-week dose-ranging studies showed that all 4 doses of cerivastatin produced significantly greater reductions in low-density lipoprotein (LDL) cholesterol than placebo. Cerivastatin 0.2 mg decreased LDL cholesterol by 30.5%. Cerivastatin also significantly decreased total cholesterol, triglycerides, and apolipoprotein B, and significantly increased high-density lipoprotein (HDL) cholesterol. Similar reductions in LDL cholesterol and total cholesterol occurred with 0.2 mg/day cerivastatin in the dose scheduling study, although the reductions were significantly greater when cerivastatin was administered once daily with either the evening meal or at bedtime compared with 2 divided doses. LDL cholesterol reductions were similar when cerivastatin was taken with the evening meal and at bedtime. Cerivastatin was well tolerated, with the incidence of adverse events comparable to that of placebo treatment. No clinically significant increases in either hepatic isoenzymes or creatine phosphokinase were observed after treatment with cerivastatin. PMID- 9737644 TI - Clinical efficacy and safety of cerivastatin: summary of pivotal phase IIb/III studies. AB - Cerivastatin is a new, third-generation 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor ("statin"), which is administered to hypercholesterolemic patients at doses equivalent to 1-3% of the doses of other statins. This report reviews the pivotal Phase IIb/III clinical trials in which the efficacy and safety of cerivastatin was compared with placebo and active comparator statins (lovastatin, simvastatin, and pravastatin) after both short- and long-term administration. Overall, the studies showed that at doses of 0.025 0.4 mg/day, cerivastatin produced dose-dependent reductions in low-density lipoprotein (LDL) cholesterol and total cholesterol, which were significantly greater than placebo. The greatest reductions were achieved with 0.4 mg/day cerivastatin. On this dose, >40% of patients achieved reductions in LDL cholesterol >40% and in a further 9% of patients, LDL cholesterol was decreased by >50%. At higher doses, cerivastatin also demonstrated potent triglyceride lowering effects in a subgroup of patients with raised plasma triglycerides. Reductions in atherogenic lipids and lipoproteins were accompanied by significant increases in high-density lipoprotein (HDL) cholesterol, apolipoprotein A-I, and antiatherogenic lipoprotein A-I. Long-term administration of cerivastatin for periods of up to 2 years was associated with persistent reductions in LDL cholesterol, total cholesterol, triglycerides, and apolipoprotein B as well as increases in HDL cholesterol similar to those observed after initial administration. Long-term cerivastatin treatment was also well tolerated. There was no significant difference between the incidence of adverse effects with cerivastatin and comparator statins or between cerivastatin and other statins with respect to clinically significant increases in either hepatic enzymes or creatine phosphokinase. In conclusion, these studies indicate that cerivastatin is a safe and effective long-term treatment for patients with primary hypercholesterolemia and also suggest that higher doses should be investigated. PMID- 9737645 TI - Cerivastatin in primary hyperlipidemia: a multicenter analysis of efficacy and safety. AB - Cerivastatin, a novel, synthetic, and enantiomerically pure 3-hydroxy-3 methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor, has been administered, in clinical trials, to >2,700 patients with primary hypercholesterolemia, of whom > 1,000 received treatment for periods of up to 1 year. A global, pooled analysis of the efficacy, safety, and tolerability of cerivastatin was performed on data obtained from all randomized, double-blind studies in which cerivastatin at doses of 0.025-0.4 mg/day was compared with either placebo or active comparator. All studies had a 10-week, diet-controlled run-in period, the last 6 weeks of which included administration of single-blind placebo. Efficacy analysis of the pooled data at 8 weeks postrandomization showed that in comparison with placebo, cerivastatin achieved significant dose-dependent reductions in low-density lipoprotein (LDL) cholesterol, the primary efficacy parameter, of 14.2-36.1 %. Reductions in LDL cholesterol were accompanied by significant reductions in total cholesterol and triglycerides, together with increases in high-density lipoprotein (HDL) cholesterol. The magnitude of the reduction in plasma triglycerides was strongly related to baseline triglyceride levels. In patients with baseline plasma triglycerides of >250 mg/dL, treatment with 0.4 mg/day cerivastatin decreased these levels by 37%. Cerivastatin was well tolerated, with the type and incidence of clinical adverse effects comparable to that of placebo and comparator drugs. The incidence of biochemical adverse effects was also similar to that seen with either placebo or comparator drugs and was independent of the dose of cerivastatin. Less than 1% of patients treated with cerivastatin at doses of 0.025-0.4 mg/day experienced clinically significant increases in either hepatic transaminases (>3x the upper limit of normal) or creatine phosphokinase (>5x the upper limit of normal). The good tolerability of cerivastatin was reflected in a low rate of premature withdrawal from treatment, below or comparable to that of placebo-treatment. The pooled efficacy and safety analyses have shown that at doses equal to 1-3% of the doses of other statins, cerivastatin is a safe, well-tolerated, and highly effective HMG-CoA reductase inhibitor for the treatment of type IIa (triglycerides <250 mg/dL) and IIb (triglycerides >250 mg/dL) hypercholesterolemia. PMID- 9737646 TI - Cerivastatin in the treatment of mixed hyperlipidemia: the RIGHT study. The Cerivastatin Study Group. Cerivastatin Gemfibrozil Hyperlipidemia Treatment. AB - The Cerivastatin Gemfibrozil Hyperlipidemia Treatment (RIGHT) study--a multicenter, randomized, double-blind, placebo-controlled study--compared the lipid-lowering effects of cerivastatin, once daily at doses of 0.1, 0.2, and 0.3 mg with those of twice-daily gemfibrozil 600 mg in 751 patients with primary mixed hyperlipidemia. Randomization to the first 16 weeks of treatment followed an initial 4-week washout period and subsequent 6-week diet-controlled, placebo run-in phase. Patients continued to receive study medication for a further 36 weeks, with those previously on placebo switched to 0.1 mg/day cerivastatin at the end of week 16. Additional cholestyramine therapy was permitted at week 36 in patients with uncontrolled low-density lipoprotein (LDL) cholesterol levels. Cerivastatin achieved significant dose-dependent reductions in LDL cholesterol of 15-24% after 16 weeks of treatment, compared with reductions of 7.5% with gemfibrozil. Over this period both cerivastatin (0.3 mg) and gemfibrozil (1,200 mg) significantly decreased levels of triglycerides (20.3% vs 50.3%, respectively) and very low-density lipoprotein (VLDL) cholesterol (30.8% vs 47.1%, respectively), as well as increasing high-density lipoprotein (HDL) cholesterol (11.3% vs 13.3%, respectively). The reductions in LDL cholesterol and other atherogenic lipids and lipoproteins at 16 weeks were sustained in the subsequent 36-week double-blind continuation phase, during which time <10% of patients received additional cholestyramine therapy. Both study drugs were well tolerated, with the incidence of adverse events similar to that of placebo treatment. Clinically significant increases in hepatic transaminases and creatine phosphokinase occurred at a similar low frequency of around 1%. This study demonstrated that cerivastatin is a safe, well-tolerated, and effective treatment for lowering elevated LDL cholesterol and triglycerides in patients with mixed hyperlipidemia. PMID- 9737647 TI - Clinical efficacy and safety of cerivastatin in the treatment of heterozygous familial hypercholesterolemia. AB - Patients with heterozygous familial hypercholesterolemia are at especially high risk of premature coronary artery disease and usually require aggressive long term lipid-lowering drug therapy to decrease plasma low-density lipoprotein (LDL) cholesterol concentrations to normal levels. In the present study, the lipid lowering effects of cerivastatin in combination with cholestyramine and probucol were investigated in 20 patients with heterozygous familial hypercholesterolemia over a 20-week treatment period. After an initial 4-week treatment with once daily 0.2 mg cerivastatin, serum total cholesterol and LDL cholesterol levels had decreased by a significant 22% and 25%, respectively (p <0.01). The addition of 8 g/day cholestyramine or 1 g/day probucol to ongoing cerivastatin therapy produced further significant reductions in total cholesterol of 16% and 16%, respectively, and in LDL cholesterol of 22% and 15%, respectively (p <0.01), over the 12-week combination therapy period. The potent lipid-lowering effects of combined treatment were accompanied by excellent toleration of study drugs. Only 2 patients experienced gastrointestinal side effects associated with cholestyramine therapy. There was no evidence of any abnormalities in creatine phosphokinase in either treatment group and only 2 patients exhibited minor increases in hepatic transaminases. This study has shown that cerivastatin can be safely combined with either cholesytramine or probucol to provide a safe and highly effective hypolipidemic treatment regimen for patients with heterozygous familial hypercholesterolemia. PMID- 9737648 TI - Perspectives and controversies in atrial fibrillation. AB - Atrial fibrillation (AF) is the most common sustained arrhythmia in humans. The 3 basic tenets of therapy are (1) restoration and maintenance of sinus rhythm; (2) ventricular rate control; and (3) prevention of thromboembolism. Maintenance of sinus rhythm appears preferable to rate control alone in patients with significant symptoms caused by AF. Complete suppression of AF with drug therapy for >6 months is unusual, but it is not the sole criterion of success. As with other chronic cardiac disorders such as angina and heart failure, a marked reduction in frequency and duration of episodes of AF will likely translate into an excellent clinical outcome. The major risk of antiarrhythmic drug therapy is ventricular proarrhythmia, which is seen most frequently in patients with substantial left ventricular dysfunction. Torsade de pointes is the most frequent proarrhythmia that occurs with antiarrhythmic agents that prolong ventricular repolarization and the QT interval. To minimize the risk of proarrhythmia, antiarrhythmic drugs are started in-hospital in patients with significant heart disease, and agents are selected based on certain patient characteristics. For example, the drugs initially selected for patients with heart failure and coronary artery disease are amiodarone and sotalol, respectively. Two approaches may be used to decrease the thromboembolic risk associated with cardioversion of AF to sinus rhythm. In the conventional method, warfarin is given (INR 2.0-3.0) for 3 weeks before and at least 4 weeks after cardioversion. An alternative approach employs transesophageal echocardiography to rule out left atrial thrombi before cardioversion. Both methods appear reasonable and safe, and I prefer the conventional and transesophageal echocardiography-guided approaches for outpatients and in-hospital patients, respectively. PMID- 9737649 TI - Measures of drug efficacy in treating atrial fibrillation. AB - Atrial fibrillation (AF) tends to recur in at least one half of the patients being treated with an antiarrhythmic agent. However, if the patient being treated is appropriately anticoagulated, and if during recurrence the ventricular response rate is controlled, a recurrence rarely requires emergent treatment. Rather, the AF recurrence can most often simply be considered a clinically important nuisance. Therefore, recurrence of AF during antiarrhythmic drug therapy should not be considered drug failure per se. Instead, the frequency and severity of recurrence constitute the measure of efficacy. Occasional recurrence may be clinically acceptable and preferable to persistent AF or frequent episodes of paroxysmal AF. Because occasional recurrence of AF should not be considered failure of drug therapy, simple outpatient cardioversion of the AF with restoration of sinus rhythm is recommended should recurrence occur and the AF persist. Simple therapies, such as acute cardioversion to restore sinus rhythm, should be considered a part of the expected long-term therapy of the patient with AF. Finally, the drug of choice should be one that is well tolerated, easy to take (preferably only once or twice a day), and has the fewest potential side effects, particularly serious adverse effects. PMID- 9737650 TI - Importance of beta blockade in the therapy of serious ventricular arrhythmias. AB - Beta blockers have traditionally been considered relatively poor antiarrhythmic agents for patients with ventricular arrhythmias. This view is based on the observations that beta blockers are less effective in suppressing spontaneous ventricular ectopy or inducible ventricular arrhythmias than are the class I and class III agents. However, there are convincing data that beta blockers can have a clinically important antiarrhythmic effect and prevent arrhythmic and sudden death. Beta blockers have multiple potential effects that can contribute to a therapeutic antiarrhythmic action, including an antiadrenergic/vagomimetic effect, a decrease in ventricular fibrillation threshold, and prevention of a catecholamine reversal of concomitant class I/III antiarrhythmic drug effects. Postinfarction trials, recent congestive heart failure studies, and observations in patients who are at risk for sustained ventricular arrhythmias all suggest a potent antiarrhythmic effect of beta blockade. PMID- 9737651 TI - Modulation of antiarrhythmic drug effects by beta-adrenergic sympathetic stimulation. AB - Appreciation has grown for the impact of the autonomic nervous system on the development of clinical cardiac arrhythmias. Antiarrhythmic medications work to significantly prolong cardiac repolarization and slow conduction. The question has arisen whether these pharmacologic actions of antiarrhythmic drugs can be modulated by alterations in the sympathetic nervous system. This article examines the data pertaining to modulation of the class I and class III effects of antiarrhythmic drugs during beta-adrenergic stimulation, the body's natural response to stress. The actions of several antiarrhythmic drugs can be fully reversed during beta-adrenergic sympathetic stimulation. Overall, the data suggest that pure class III drugs are the most susceptible to reversal of their effects on refractoriness, followed by class IA agents, amiodarone (which has partial resistance), and d,l-sotalol (which is highly resistant to reversal). Whereas retrospective analyses of a number of trials suggest that sympathetic stimulation-induced reversal of the electrophysiologic effects of certain antiarrhythmic drugs can decrease their clinical efficacy, prospective trials examining this issue are needed. At the current time it appears reasonable to administer beta blockers to patients receiving antiarrhythmic agents that do not have intrinsic antiadrenergic effects. PMID- 9737652 TI - Antiarrhythmic drugs and devices for the management of ventricular tachyarrhythmia in ischemic heart disease. AB - Ischemic heart disease is the most frequent cardiac abnormality in patients with sustained or nonsustained ventricular tachyarrhythmias. The goals of therapy in such patients are to decrease the severity and incidence of symptoms and prolong life. In this article, we review the current views on antiarrhythmic drug therapy and an implantable cardioverter-defibrillator (ICD) in patients with ischemic heart disease. The importance of beta blockade as part of the therapy is emphasized. In addition, the superiority of sotalol and amiodarone over class I drugs, the benefits of combined treatment with amiodarone and a beta blocker, and the impact and limitations of current trials comparing the effectiveness of drug therapy with that of an ICD are all considered. Also discussed is the combined use of an antiarrhythmic drug and an ICD. In this approach sotalol is generally the agent of choice, with amiodarone the second choice. PMID- 9737653 TI - Interactions between implantable cardioverter-defibrillators and class III agents. AB - Although implantable cardioverter-defibrillators (ICDs) can successfully terminate ventricular arrhythmias, antiarrhythmic drugs are often required to prevent recurrent events. Class III antiarrhythmic agents have emerged as the safest, most effective, and widely used agents in the 40-70% of ICD patients who require concomitant antiarrhythmic medication. Antiarrhythmic agents can influence the effectiveness of ICDs to terminate arrhythmias through their effect on defibrillation threshold. All class III agents share the ability to prolong ventricular refractoriness and those with "pure" class III activity consistently decrease defibrillation threshold in the normal canine heart model. Sotalol, amiodarone, and bretylium all have other Vaughan Williams class actions that influence their respective effects on defibrillation threshold. Sotalol has been associated with a decrease in defibrillation threshold in both animal and in clinical studies, whereas amiodarone has been associated with variable effects in animal models and an increase in defibrillation threshold in clinical studies. Additionally, antiarrhythmic agents may prolong ventricular tachycardia (VT) cycle length, which may affect the ability to pace terminate or cardiovert VT. Amiodarone has a moderate slowing effect on the VT cycle length. Finally, class III drugs also have proarrhythmic potential that may affect the defibrillator's function. Sotalol can be associated with dose-related torsade de pointes, whereas amiodarone may slow the VT cycle length below the tachycardia detection rate cutoff. In conclusion, class III pharmacotherapy can be safely administered in conjunction with ICD therapy as long as the interaction between these therapeutic modalities is appreciated. PMID- 9737654 TI - Mechanisms and management of proarrhythmia. AB - It is now well recognized that therapy with antiarrhythmic drugs can not only suppress cardiac arrhythmias, but also may increase their frequency or provoke new ones. Specific proarrhythmia syndromes, each with a distinct underlying mechanism and approach to therapy, have been described. The best-recognized examples are digitalis intoxication, proarrhythmia associated with sodium-channel block, and torsade de pointes occurring during QT-prolonging therapies. In the case of sodium-channel blockers, 2 forms of proarrhythmia are commonly recognized: slow atrial flutter with 1:1 atrioventricular conduction, and frequent ventricular tachycardia ([VT], most often found in patients with pre existing VT reentrant circuits). In all cases, the best approach to therapy is to identify patients at risk (and thereby avoid therapy entirely), to recognize proarrhythmia when it occurs, to withdraw offending agent(s), and to use specific corrective therapies when available. Although most recognized episodes of proarrhythmia are thought to occur early in drug therapy, the increased mortality during chronic antiarrhythmic therapy demonstrated in large randomized trials suggests this phenomenon can also develop during long-term drug treatment. The recognition of proarrhythmia and the delineation of its underlying mechanisms should not only improve therapy with available drugs, but may also direct development of newer agents devoid of this potential. PMID- 9737655 TI - Antiarrhythmic drug therapy in pregnancy and lactation. AB - Antiarrhythmic agents commonly used in clinical practice are reviewed with respect to their potential for teratogenic or other adverse fetal effects. Although most experience with antiarrhythmic drug therapy during pregnancy has accrued with digoxin, quinidine, and propranolol, other antiarrhythmic agents may also be used in the pregnant patient if indicated. The choice of antiarrhythmic drug depends on the specific arrhythmia being treated, the cardiac condition of the patient or fetus, and the known or anticipated actions of the antiarrhythmic drug being considered. The management of specific arrhythmias encountered in pregnant women are also discussed. For benign arrhythmias, a conservative approach starting first with preventive measures is appropriate. For more severe or symptomatic arrhythmias, pharmacologic therapy should be instituted using drugs with proven safety to the fetus, if possible. Electrical cardioversion of the patient may be performed with relative safety in more emergent situations. PMID- 9737656 TI - IL-4: role in disease and regulation of production. PMID- 9737658 TI - Autoantibodies to C-reactive protein (CRP) and other acute-phase proteins in systemic autoimmune diseases. AB - Autoantibodies to CRP were reported previously in patients suffering from toxic oil syndrome. This syndrome resembles autoimmune diseases such as systemic lupus erythematosus (SLE) or systemic scleroderma. We therefore examined the prevalence of antibodies to CRP and other acute-phase proteins in autoimmune diseases, including SLE, subacute cutaneous lupus erythematosus (SCLE), systemic scleroderma (SSc), and primary biliary cirrhosis (PBC), as well as in bone marrow transplantation-induced chronic graft-versus-host disease and eosinophilia myalgia syndrome. IgG antibodies to CRP were found in 78% of SLE and in 30% of SCLE patients, while 16% of patients with PBC were positive. In up to 45% of patients with SSc predominantly IgG antibodies to ceruloplasmin were detectable. Lack of systemic involvement as in discoid lupus erythematosus and localized scleroderma (morphea) correlated with low or absent antibody formation. However, no correlation was found between anti-acute-phase protein antibodies with liver disease or other organ involvement. Adsorption studies revealed that non-native epitopes on the CRP molecule, termed modified CRP, are the main target of antibodies. Chronic inflammatory tissue injury in systemic autoimmune disease might increase the presentation of cryptic epitopes of CRP to the threshold required for T cell activation. PMID- 9737657 TI - Effects of anti-IL-4 receptor monoclonal antibody on in vitro T cell cytokine levels: IL-4 production by T cells from non-atopic donors. AB - IL-4 is a pleiotropic cytokine which is involved in the development of atopic diseases. Only limited data exist on IL-4 production in humans, and the relative contribution to atopy of either unbalanced IL-4 production, or increased IL-4 responsiveness of target cells, is still unknown. The use of a MoAb to the IL-4 receptor alpha-chain (IL-4Ralpha) enabled us to demonstrate that IL-4 production in vitro is usually underestimated, due to in vitro consumption, even in cultures of purified T cells. When IL-4 consumption was blocked, it became evident that CD80 and CD86 both provide effective costimulatory signals for high IL-4 production. Moreover, we found that even stimulation with a soluble antigen (tetanus toxoid) induces IL-4 production by T cells from healthy non-atopic donors. Both sets of data imply that IL-4 is not required for IL-4 production by memory and/or effector T cells. Our data further show that endogenous IL-4 activity modulates IL-10 and interferon-gamma production by T cells in opposite directions. The use of this receptor-blocking antibody will thus be helpful for in vitro studies on IL-4 regulation. Consumption of IL-4 by different cell types during in vitro cultures might have interfered with previous attempts to quantify IL-4 production by human T cells. PMID- 9737659 TI - A dominant role for non-MHC gene effects in susceptibility to cyclosporin A (CsA) induced autoimmunity. AB - Lethally irradiated LEW rats reconstituted with syngeneic bone marrow and given CsA for a 4-week period develop a graft-versus-host-like disease upon withdrawal of CsA. This T cell-mediated autoimmune disease is referred to as CsA-induced autoimmunity (CsA-AI). CsA-AI-susceptible LEW rats and resistant BN rats differ greatly in the composition of their peripheral T cell compartment. To dissect the role of MHC and non-MHC genes in the development of peripheral T cell subsets in combination with susceptibility to CsA-AI the respective MHC congenic strains (LEW-1N and BN-1L) were examined for their T cell subsets and for their ability to develop CsA-AI. In this study we show that the Th1/Th2-like cell ratio as well as susceptibility to CsA-AI are under control of the non-MHC genes. This suggests that the Th1/Th2-like cell ratio is a critical determinant for development of CsA AI. Alternatively, resistance can be attributed to lack of target organ susceptibility due to the absence of the target autoantigen in resistant rat strains. This interpretation is rejected, since both BN as well as BN-1L rats consistently develop the characteristic macroscopic and microscopic signs of CsA AI upon adoptive transfer with autoreactive LEW-1N and LEW T cells, respectively. Therefore, it can be concluded that the non-MHC genes encode for immune deviation and thereby determine susceptibility or resistance to CsA-AI. PMID- 9737660 TI - Study of antigenic sites on the asialoglycoprotein receptor recognized by autoantibodies. AB - The aim of this study was to identify the epitopes recognized by antibodies to the asialoglycoprotein receptor, a specific hepatocyte protein, from sera of patients with autoimmune hepatitis. An ELISA test was used to detect anti asialoglycoprotein receptor antibodies in the sera of patients with autoimmune hepatitis. Positive sera were tested against the same antigen by slot blot, by Western blot and by immunoprecipitation of the untreated protein and following treatment with beta-mercaptoethanol (beta-ME) and endoglycosidase F. The mature, unglycosylated and partially glycosylated forms of the asialoglycoprotein receptor synthesized by HepG2 cells were tested against positive patients' sera, as well as the in vitro translated unglycosylated form of the H1 subunit of the receptor. Sera from patients with autoimmune hepatitis recognized equally the native form, as well as the beta-ME-modified form, but less well the deglycosylated form of the human mature receptor. No reactivity was found when these sera were tested against the denatured human protein. In addition, neither the unglycosylated H1 subunit nor any of the HepG2-synthesized asialoglycoprotein receptor forms bound to the antibodies. Altogether, these results show that anti asialoglycoprotein receptor antibodies in the sera of patients with autoimmune hepatitis are directed against conformational structures of the mature hetero oligomeric form of the human liver protein and that at least some epitopes were located on the extracellular domain of the antigen. PMID- 9737661 TI - Pattern of expression of tetraspanin antigen genes in Burkitt lymphoma cell lines. AB - Tetraspanin antigens are implicated in the prognosis of different types of tumours. In this study we determine by semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR) the level of 13 tetraspan messages in 21 Burkitt lymphoma (BL) cell lines. All tumour cell lines have a common pattern of tetraspanin gene expression. There are five antigens which are detected in 90% of cell lines at high levels, CD53, CD81, CD63, SAS and CD82. Another two, CD9 and CD37, were detected in 60% of cell lines, and have a very variable level of expression. The remaining antigens, A15, CoO29, KRAG, L6, TI-1 and il-TMP, are expressed at low levels in very few cell lines without any specific pattern. The level of gene expression corresponds with the level of cell surface antigen determined by flow cytometry. The average number of tetraspan proteins expressed per cell line is six. These proteins may form subunits of an oligomeric structure with 24 transmembrane domains. There are no major differences in tetraspan expression pattern among sporadic or endemic tumours, type of translocation or Epstein-Barr virus status, suggesting the original cell of these tumours is the same, probably a late pre-B cell, at the CD9 to CD37 transition point. Tetraspanin gene expression is consistent with BL being a single entity, despite variations in other parameters. PMID- 9737663 TI - Origin and properties of soluble CD21 (CR2) in human blood. AB - By analysis with a panel of CD21 MoAbs it is shown that a large part of the soluble CD21 in human blood plasma is of the long isoform (CD21L), as judged by comparison with antigen produced by mouse L cells transfected with CD21L-cDNA and reactivity with the restricted CD21 MoAb R4/23. This is compatible with the hypothesis that soluble CD21 in the blood is mainly derived from follicular dendritic cells (FDC). Cells from a human keratinocyte cell line transfected with cDNA from the Burkitt lymphoma cell line Raji also produced soluble CD21L (sCD21L), whereas the short form of sCD21 (sCD21S) was the major component of sCD21 produced by the B lymphoblastoid cell line LICR-LON-HMy and the T cell line Jurkat. Confocal studies of FDC isolated from human tonsil revealed that CD21 was present in the cytoplasm. On gel filtration sCD21 from untreated serum has an apparent size considerably greater than the 130kD found by SDS-PAGE analysis. This may be partly accounted for by the non-globular shape of the molecule, but may also indicate, as reported by others, that in its native state sCD21 is complexed with other proteins. However, no evidence of complexing with sCD23 or C3d could be found. PMID- 9737662 TI - Complement regulation in innate immunity and the acute-phase response: inhibition of mannan-binding lectin-initiated complement cytolysis by C-reactive protein (CRP). AB - Mannan-binding lectin (MBL) is an acute-phase protein which activates complement at the level of C4 and C2. We recently reported that the alternative pathway also is required for haemolysis via this 'lectin pathway' in human serum. CRP is another acute-phase reactant which activates the classical pathway, but CRP also inhibits the alternative pathway on surfaces to which it binds. Since serum levels of both proteins generally increase with inflammation and tissue necrosis, it was of interest to determine the effect of CRP on cytolysis via the lectin pathway. We report here that although CRP increases binding of C4 to MBL sensitized erythrocytes, which in turn enhances lectin pathway haemolysis, it inhibits MBL-initiated cytolysis by its ability to inhibit the alternative pathway. This inhibition is characterized by increased binding of complement control protein H and decreased binding of C3 and C5 to the indicator cells, which in turn is attributable to the presence of CRP. Immunodepletion of H leads to greatly enhanced cytolysis via the lectin pathway, and this cytolysis is no longer inhibited by CRP. These results indicate that CRP regulates MBL-initiated cytolysis on surfaces to which both proteins bind by modulating alternative pathway recruitment through H, pointing to CRP as a complement regulatory protein, and suggesting a co-ordinated role for these proteins in complement activation in innate immunity and the acute-phase response. PMID- 9737664 TI - Expression, characterization, processing and immunogenicity of an insulin dependent diabetes mellitus autoantigen, IA-2, in Sf-9 cells. AB - Autoantibodies to a 64-kD protein and a 40-kD tryptic fragment from pancreatic islets have been detected at high frequency in the sera of patients with insulin dependent diabetes mellitus (IDDM). IA-2, a newly isolated transmembrane protein tyrosine phosphatase, is a major islet cell autoantigen in IDDM and the precursor of a 40-kD tryptic fragment. To express large quantities of recombinant IA-2 protein and analyse post-translational modifications we expressed full-length human IA-2 in baculovirus-infected Sf-9 cells. IA-2 expression was analysed by Western blot and by immunoprecipitation of 35S-methionine-radiolabelled proteins with rabbit antisera or IDDM sera. A 120-kD IA-2 protein was detected during the early, but not the late, phase of the infection. Pulse-chase experiments showed that the 120-kD protein was processed into fragments of 64 kD and smaller fragments of approximately 50 kD, 38 kD and 32 kD. The 64-kD fragment appeared as a doublet. Tunicamycin and PNGase F treatment down-shifted the 120-kD protein and the 64-kD doublet into lower molecular weight bands, suggesting that both were glycosylated. Trypsin treatment converted the 120-kD protein and the 64-kD doublet into a 40-kD fragment. Baculovirus-expressed IA-2 was as sensitive or slightly more sensitive than in vitro translated IA-2 in detecting autoantibodies to IA-2: 66% of sera from newly diagnosed IDDM patients reacted with baculovirus expressed IA-2 compared with 59% of the same sera which reacted with in vitro translated IA-2. It is concluded that baculovirus-expressed IA-2 is a good source of autoantigen and that a number of lower molecular weight fragments with which IDDM autoantibodies react are derived from the 120-kD full-length IA-2 molecule. PMID- 9737665 TI - Autoimmune thrombocytopenia (AITP) and thyroid autoimmune disease (TAD): overlapping syndromes? AB - The pathogenesis of thrombocytopenia associated with TAD and the occurrence of overlapping traits between TAD and AITP are still a matter of debate. For this reason, we investigated for the presence and specificity of platelet and thyroid autoantibodies in 18 TAD patients with thrombocytopenia, 19 TAD patients without thrombocytopenia and in 22 patients with primary AITP without clinical signs of TAD. Platelet-associated IgG and/or specific circulating platelet autoantibodies were detected in 83% of patients with TAD and thrombocytopenia, in 10% of patients with TAD without thrombocytopenia and in 86% of patients with primary AITP. The reactivity of serum autoantibodies, assayed by MoAb immobilization of platelet antigens (MAIPA), was directed against platelet glycoproteins Ib and/or IIb/IIIa in 50% of the patients with TAD and thrombocytopenia, as in 46% of the patients with primary AITP. Thyroid autoantibodies were found in 89% of patients with TAD and thrombocytopenia, in 95% of patients with TAD without thrombocytopenia, and in 18% of patients with primary AITP. Thyrotropin (TSH) levels determined in three of four AITP patients with thyroid autoantibodies revealed a subclinical hyperthyroidism in one patient. The present study supports the autoimmune aetiology of thrombocytopenia associated with TAD, since the prevalence and specificity of platelet autoantibodies are similar in TAD and primary AITP. The results indicate also that there exists an overlap between thyroid and platelet autoimmunity with or without clinical manifestations. PMID- 9737666 TI - Cytokine-stimulated release of decay-accelerating factor (DAF;CD55) from HT-29 human intestinal epithelial cells. AB - Expression of DAF (CD55) is enhanced on colonic epithelial cells of patients with ulcerative colitis (UC), and stool DAF concentrations are increased in patients with active disease. Cytokines are known to modulate DAF expression in various human cells, and lesions of UC reveal altered profiles of cytokine production. In this study, we evaluate the effects of various cytokines, IL-1beta, IL-2, IL-4, IL-6, IL-8, IL-10, and interferon-gamma (IFN-gamma), on the synthesis and kinetics of DAF protein in HT-29 human intestinal epithelial cells. Using flow cytometry and an ELISA, we found that HT-29 cells constitutively express DAF on the cell surface and spontaneously release DAF into the culture supernatant under standard culture conditions. When the culture supernatant was centrifuged at 100000g, nearly a half of DAF was precipitated, indicating that one half of the released DAF was present as a membrane-bound form and the other half as a soluble form. Analysis of the culture supernatant of biotin surface-labelled HT-29 cells suggested that the soluble form DAF was derived by secretion from within the cell or by cleavage from the cell surface. Among the cytokines, IL-4 markedly, and IL 1beta moderately, enhanced the expression and the release of DAF. Actinomycin D, cycloheximide, and brefeldin A inhibited the increase in DAF release induced by IL-4 and IL-1beta stimulation. These results suggest that DAF is released from intestinal epithelial cells in response to cytokine stimulation and that IL-4 and IL-1beta are possible cytokines involved in DAF release into the colonic lumen of patients with UC. PMID- 9737667 TI - The murine buccal mucosa is an inductive site for priming class I-restricted CD8+ effector T cells in vivo. AB - The present study shows that Langerhans cells of the buccal mucosa and the skin share a similar phenotype, including in situ expression of MHC class II, the mannose receptor DEC-205 and CD11c, and absence of the costimulatory molecules B7.1, B7.2 and CD40 as well as Fas. Application of 2,4-dinitrofluorobenzene (DNFB) onto the buccal mucosa is associated with a rapid migration of dendritic cells (DC) to the epithelium and induction of B7.2 expression on some DC. Buccal sensitization with DNFB elicited a specific contact sensitivity (CS) in response to skin challenge, mediated by class I-restricted CD8+ effector T cells and down regulated by class II-restricted CD4+ T cells, demonstrated by the lack of priming of class I-deficient mice and the enhanced response of class II-deficient mice, respectively. CS induced by buccal immunization is associated with priming of class I-restricted CD8+ effector T cells endowed with hapten-specific cytotoxic activity. Thus, the buccal epithelium is an inductive site, equivalent to the epidermis, for the generation of CS independent of CD4 help, and of cytotoxic T lymphocyte (CTL) responses mediated by class I-restricted CD8+ T cells. We propose that immunization through the buccal mucosa, which allows antigen presentation by epithelial DC efficient for priming systemic class I restricted CD8+ CTL, may be a valuable approach for single-dose mucosal vaccination with subunit vaccines. PMID- 9737668 TI - Differential effect on TCR:CD3 stimulation of a 90-kD glycoprotein (gp90/Mac 2BP), a member of the scavenger receptor cysteine-rich domain protein family. AB - We studied the effects of a 90-kD glycoprotein (gp90/Mac-2BP) belonging to the scavenger receptor family, present in normal serum and at increased levels in inflammatory disease and cancer patients, on some T cell function parameters. Whereas the lymphocyte proliferative response to non-specific mitogens such as phytohaemagglutinin (PHA) and concanavalin A (Con A), but not pokeweed mitogen (PWM), was strongly reduced, probably due to the lectin-binding properties of gp90/Mac-2BP, the response to T cell receptor (TCR) agonists such as superantigens and allogeneic cells was potentiated. When lymphocytes were stimulated with different anti-TCR:CD3 MoAbs, both in soluble and solid-phase form, gp90/Mac-2BP was able to down-regulate the proliferative response to anti CD3 MoAb, whereas the response to anti-TCR alphabeta MoAb was enhanced. A similar differential effect was observed when a MoAb against CD5 (another member of the scavenger receptor superfamily) was added to anti-CD3 or anti-TCR-stimulated cells; anti-CD5 MoAb strongly down-modulated the CD3-mediated response, whereas its presence in culture was associated with potentiation of the response to TCR alphabeta agonists. gp90/Mac-2BP was able per se to up-regulate Ca2+ levels in freshly isolated lymphocytes; moreover, its presence in culture was associated with increased Ca2+ mobilization following stimulation with anti-TCR alphabeta, but not anti-CD3 MoAb. These data indicate that gp90/Mac-2BP could be able to influence some immune responses, possibly through multiple homologous interactions with other members of the scavenger receptor family; moreover, our findings suggest that signalling through the different components of the TCR:CD3 complex may follow distinct activation pathways into the cells. PMID- 9737669 TI - Tumour necrosis factor (TNF) gene polymorphism influences TNF-alpha production in lipopolysaccharide (LPS)-stimulated whole blood cell culture in healthy humans. AB - TNF-alpha is involved in infectious and immuno-inflammatory diseases. Different individuals may have different capacities for TNF-alpha production. This might determine a predisposition to develop some complications or phenotypes of these diseases. The aims of our study were to assess the inter-individual variability of TNF-alpha production and to correlate this variability to a single base pair polymorphism located at position -308 in TNF gene. We studied 62 healthy individuals. TNF-alpha production after LPS stimulation was evaluated using a whole blood cell culture model. The TNF gene polymorphism was studied by an allele-specific polymerase chain reaction. Other cytokines produced in the culture, soluble CD14 concentrations and expression of CD14 on blood cells were also measured. Among the 62 individuals, 57 were successfully genotyped. There were 41 TNF1 homozygotes and 16 TNF1/TNF2 heterozygotes. TNF-alpha production after LPS stimulation of whole blood cell culture was higher among TNF2 carriers than among TNFI homozygotes (929pg/ml (480-1473pg/ml) versus 521 pg/ ml (178-1307 pg/ml); P<0.05). This difference was even more significant after correction of TNF-alpha production for CD14 expression on blood cells. In conclusion, the single base pair polymorphism at position -308 in the TNF gene may influence TNF alpha production in healthy individuals. PMID- 9737670 TI - A variety of cytokines and immunologically relevant surface molecules are expressed by normal human skeletal muscle cells under proinflammatory stimuli. AB - Muscle is an attractive target for gene therapy and for immunization with DNA vaccines and is also the target of immunological injury in myositis. It is important therefore to understand the immunologic capabilities of muscle cells themselves. In this study, we show that proinflammatory stimuli induce the expression of other cytokines such as IL-6, transforming growth factor-beta (TGF beta), and granulocyte-macrophage colony-stimulating factor (GM-CSF) by muscle cells themselves, as well as the up-regulation of human leucocyte antigen (HLA) class I, class II and intercellular adhesion molecule-1 (ICAM-1). Thus, muscle cells have an inherent ability to express and respond to a variety of cytokines and chemokines. The levels of HLA class I, class II and ICAM-1 in inflamed muscle may be affected by the secreted products of the stimulation. PMID- 9737671 TI - FcgammaRIa-gamma-chain complexes trigger antibody-dependent cell-mediated cytotoxicity (ADCC) in CD5+ B cell/macrophage IIA1.6 cells. AB - Most receptors for immunoglobulins exist as multi-subunit complexes, with unique ligand binding alpha-chains, combined with accessory signalling (gamma-, beta-, or zeta-) chains. The myeloid class I receptor for IgG (FcgammaRIa) has been shown to be dependent on the FcR gamma-chain for surface expression in vivo. In this study we assess the capacity of FcgammaRIa-gamma-chain complexes expressed in IIA1.6 cells to trigger phagocytosis and ADCC. An intact immunoreceptor tyrosine-based activation motif (ITAM) signalling motif proved essential for triggering of biological function via the FcgammaRIa receptor complex. Both the FcR gamma-chain and the FcgammaRIIa-ITAM proved active in directing phagocytosis of Staphylococcus aureus and ADCC of erythrocytes, triggered by the FcgammaRIa complex. The capacity of FcgammaRIa to trigger phagocytic and cytolytic activity by IIA1.6 cells, both considered 'professional phagocyte' functions, motivated us to re-evaluate the cell lineage and developmental stage of IIA1.6 cells. Although originally described as mouse B lymphocytes, the IIA1.6 cells proved positive for non-specific esterase activity and expressed the CD5 antigen. These combined characteristics place the IIA1.6 cells within a unique CD5+ B cell/macrophage lineage, optimally suited for cell biological analyses of phagocyte receptors. PMID- 9737672 TI - Interaction of murine macrophage-membrane proteins with components of the pathogenic fungus Histoplasma capsulatum. AB - The interaction of macrophage-membrane proteins and histoplasmin, a crude antigen of the pathogenic fungus Histoplasma capsulatum, was studied using murine peritoneal macrophages. Membrane proteins were purified via membrane attachment to polycationic beads and solubilized in Tris-HCl/SDS/DTT/glycerol for protein extraction; afterwards they were adsorbed or not with H. capsulatum yeast or lectin binding-enriched by affinity chromatography. Membrane proteins and histoplasmin interactions were detected by ELISA and immunoblotting assays using anti-H. capsulatum human or mouse serum and biotinylated goat anti-human or anti mouse IgG/streptavidin-peroxidase system to reveal the interaction. Results indicate that macrophage-membrane proteins and histoplasmin components interact in a dose-dependent reaction, and adsorption of macrophage-membrane proteins by yeast cells induces a critical decrease in the interaction. Macrophage-membrane glycoproteins with terminal D-galactosyl residues, purified by chromatography with Abrus precatorius lectin, bound to histoplasmin; and two bands of 68kD and 180kD of transferred membrane protein samples interacted with histoplasmin components, as revealed by immunoblot assays. Specificity for beta-galactoside residues on the macrophage-membrane was confirmed by galactose inhibition of the interaction between macrophage-membrane proteins and histoplasmin components, in competitive ELISA using sugars, as well as by enzymatic cleavage of the galactoside residues. PMID- 9737673 TI - In vivo tracking and protective properties of Yersinia-specific intestinal T cells. AB - After invasion via M cells enteropathogenic Yersinia enterocolitica subsequently establish an infection at three different sites: (i) Peyer's patches (PP), (ii) mesenteric lymph nodes (MLN), and after systemic dissemination in (iii) spleen, liver and lung. In order to characterize protective properties of intestinal T cells at the different sites of Y. enterocolitica infection, PP and MLN T cells were isolated from Y. enterocolitica-infected C57B1/6 mice and Yersinia-specific T cell lines were generated. These T cells exhibited the phenotype of CD4 Th1 cells. The adoptive transfer of Yersinia-specific Th1 cells from PP and MLN conferred protection against a lethal orogastric inoculum with Y. enterocolitica as revealed by survival post-infection. However, determination of bacterial counts in infected organs revealed that the transfer of PP T cells conferred protection in spleen but not in MLN and PP, whereas the transfer of T cells from MLN reduced bacterial counts in both spleen and MLN but not in PP. To elucidate the different protection pattern we wanted to track the transferred cells in vivo. For this purpose the cells were labelled with the stable green fluorescent cell linker PKH2-GL prior to the adoptive transfer. In vivo tracking of these cells revealed that the distribution pattern of transferred T cells in spleen, MLN and PP correlated closely with the protection pattern observed after Yersinia infection. Thus, most cells were recovered from the spleen, while only few cells were recovered from MLN and PP. In keeping with these results a rapid and significant increase in interferon-gamma (IFN-gamma) production in the spleen of mice after adoptive transfer of T cell lines was observed. Taken together, the present results demonstrate that intestinal CD4 Th1 cells from PP and MLN may be involved in the defence against Y. enterocolitica at different sites of the infection, and that PKH2-GL labelling is a suitable tool to characterize T cell functions in vivo. PMID- 9737674 TI - Immunocytochemical localization of inducible nitric oxide synthase and transforming growth factor-beta (TGF-beta) in leprosy lesions. AB - Inducible nitric oxide synthase (iNOS) and TGF-beta were localized by immunocytochemistry in skin lesions from patients across the leprosy spectrum, and from patients undergoing reversal reaction. iNOS expression was highest at the tuberculoid pole of the spectrum, and increased during reversal reaction. TGF beta was observed throughout the leprosy spectrum, but was highest at the lepromatous pole. Levels of TGF-beta decreased during reversal reaction. Reduced levels of TGF-beta may contribute to unregulated inflammatory responses during reactional episodes. PMID- 9737675 TI - Cytokine production by CD4 and CD8 T cells during the growth of Mycobacterium tuberculosis in mice. AB - Changes in the pattern of cytokines found in CD4 and CD8 T cells during the growth of Mycobacterium tuberculosis that resulted in the establishment of a latent infection were monitored. Subsets of T cells were identified based on their differential expression of CD45 and CD44 which allowed them to be classified as naive, activated or memory. We found that the T cells in the lung produced a predominantly type 1 cytokine response. The appearance of large numbers of Th1 cells coincided with the establishment of latency. In contrast, the predominant response in the mediastinal lymph node and spleen was a Th2-type response. PMID- 9737676 TI - Increased number of T cells committed to IL-5 production after respiratory syncytial virus (RSV) infection of human mononuclear cells in vitro. AB - We examined changes in the cytokine profile of T cells induced by in vitro infection with RSV. Isolated mononuclear cells from 27 healthy adults and six infants were infected with RSV at a concentration of 3 MOI (multiplicity of infection). After 48 h cells were restimulated with phorbol ester and ionomycin in the presence of monensin for 5 h. The intracellular expression of viral antigen, the cytokines IL-2, IL-4, IL-5, interferon-gamma (IFN-gamma), and the expression of surface markers were assessed by immunofluorescent staining and flow cytometry. There was a significant (P<0.001) rise of IL-5 expression in RSV infected cultures in comparison with uninfected cultures from the same individuals, whereas there were no changes in the expression of the other lymphokines. The increase in IL-5 generation depended on viable infectious RSV rather than inactivated virus. RSV infection as well as IL-5 production in T cells were confined to the CD8 subpopulation. However, there was no simultaneous expression of RSV antigen and IL-5. Purified T cells did not show any increase in IL-5 generation. However, when the rate of RSV infection was enhanced in monocytes by means of a specific monoclonal antibody, co-cultured T cells displayed an increase of IL-5 production compared with samples with ordinary low rate RSV infection. It is therefore likely that the increased commitment of lymphocytes to produce IL-5 after RSV infection in vitro is mediated by monocytes or other antigen-presenting cells. PMID- 9737677 TI - Abnormal T cell receptor V gene usage in myasthenia gravis: prevalence and characterization of expanded T cell populations. AB - The usage of T cell receptor (TCR) Valpha/Vbeta chains on cells from 38 patients with myasthenia gravis (MG) was determined by flow cytometry. There was a decreased number of cells expressing Vbeta2 in CD8+ and Vbeta3 in CD4+ cells in patients compared with healthy individuals. Abnormal expansions of T cells using particular TCR Valpha/Vbeta gene products were found in 18/38 patients. A significantly higher usage of Vbeta13 was observed but there was no restriction with regard to other TCR Valpha/Vbeta. Expanded cells belonging to both CD4+ and CD8+ were present in MG patients while restricted to the CD8+ population in healthy individuals. To elucidate the role of the expanded populations, we studied characteristics of the expanded and non-expanded T cells from MG patients who had persistent T cell expansions over more than 2 years. The cells were analysed with regard to phenotype, cytokine secretion, cytokine mRNA expression and reactivity with the autoantigen, the acetylcholine receptor. The characteristics of the expanded populations in MG clearly differed from those found in healthy individuals. More cells in the CD4+ expanded populations expressed HLA-DR and there was also a tendency for higher expression of CD25, CD28 and CD57. The number of cells spontaneously secreting cytokines was higher in the expanded populations. A dominant Th1-type cytokine secretion and mRNA expression was noted. Autoantigen-reactive CD4+ T cells were largely restricted to the expanded populations. PMID- 9737679 TI - Internal tandem duplication of the FLT3 gene is a novel modality of elongation mutation which causes constitutive activation of the product. AB - An internal tandem duplication (ITD) of the FLT3 gene is found in nearly 20% of acute myeloid leukemia (AML) and 5% of myelodysplastic syndrome cases. Our serial studies on 51 samples with the FLT3 gene mutation indicated that the ITD was frequently (47/51) clustered in the tyrosine-rich stretch from codon 589 to 599 and rarely (3/51) in its downstream region, both of which are located within the juxtamembrane (JM) domain. One remaining sample had an insertion into the JM domain of nucleotides of unknown origin. To elucidate the biological relevance of the ITD or the insertion, we expressed various types of mutant FLT3 in Cos 7 cells. All mutant FLT3 studied were ligand-independently dimerized and their tyrosine residues were phosphorylated. The Y589 of FLT3 was essential for the phosphorylation in the wild FLT3, but a Y589F conversion did not affect the phosphorylation status of the mutant FLT3. These findings suggest that the elongation of the JM domain rather than increase of tyrosine residues causes gain of-function of FLT3. Thus, ITD is a novel modality of somatic mutation which activates its product. Since the DNA corresponding to codon 593 to 602 potentially forms a palindromic intermediate, we propose that a DNA-replication error might be associated with generating the ITD of the FLT3 gene. PMID- 9737678 TI - Endogenous opioids modulate allograft rejection time in mice: possible relation with Th1/Th2 cytokines. AB - The continuous infusion of the opioid peptide beta-endorphin prolongs skin allograft survival in mice, while the opiate receptor antagonist naloxone, administered together with the opioid at the time of transplantation, abolishes the effect of the opioid. Consistently, naloxone, when given alone at the time of transplantation, but not later, accelerates graft rejection and increases splenocyte IL-2 and interferon-gamma (IFN-gamma) production. Splenocyte beta endorphin concentrations are lower in transplanted animals. The effects of exogenous beta-endorphin and naloxone suggest that the endogenous opioid peptide beta-endorphin exerts a tonic inhibitory effect over early events of T cell mediated immune responses in vivo. The effects of beta-endorphin and naloxone are consistent with the previously shown role of the opioid system in the modulation of the Th1/Th2 cytokine pattern. PMID- 9737680 TI - Antifungal prophylaxis with itraconazole in neutropenic patients with acute leukaemia. AB - The efficacy of antifungal prophylaxis with itraconazole capsules and its serum concentrations were evaluated in patients intensively treated for acute leukaemia. A consecutive group of patients without systemic antifungal prophylaxis (January 1993 to August 1994, period 1) was compared with another consecutive group of patients (period 2) who received itraconazole capsules (September 1994 to April 1995 400 mg/day, from May 1995 onwards 600 mg/day). All patients admitted with acute leukaemia and standard or high-dose chemotherapy were included into the study. Clinical endpoint was mortality from proven fungal infection. Seventy-six patients and 148 courses of cytotoxic chemotherapy were analysed in the control group as well as 47 patients and 112 treatment courses in the intervention group. Antifungal prophylaxis led to a significant decrease of mortality from invasive fungal infections (8.8%-0.9%, P = 0.005). The median trough concentration of itraconazole of all measurements was 520 ng/ml (range 230 793) in patients who received 400 mg/day and 760 ng/ml (370-1200) in patients receiving a dosage of 600 mg/day (P = 0.002). These findings suggest that itraconazole is an effective drug for antifungal prophylaxis but also that a considerable number of patients do not reach the desired trough levels (>500 ng/ml) with itraconazole capsules. PMID- 9737681 TI - Relation between age, immunophenotype and in vitro drug resistance in 395 children with acute lymphoblastic leukemia--implications for treatment of infants. AB - The prognosis of infant ALL, characterized by a high incidence of the immature CD10 negative B-lineage ALL (proB ALL) is poor. This study aimed to determine the resistance profile of infant ALL cells. In vitro drug resistance was determined by the MTT assay of 395 children with ALL at initial diagnosis: there were 21 infants <1.5 years of which nine <1 year, 284 children aged 1.5-10 years (intermediate age group) and 90 children >10 years. Immunophenotyping resulted in 310 cALL/preB ALL, 69 T-ALL, 15 proB ALL and one unknown cases. The following drugs were tested: daunorubicin, doxorubicin, mitoxantrone, idarubicin (Ida), prednisolone (Pred), dexamethasone (DXM), vincristine (VCR), Asparaginase (Asp), 6-MP, 6-TG, AraC, VM26 and 4-HOO-ifosfamide (Ifos). Infants <1.5 years were significantly more resistant to Pred (>500-fold), Asp (11-fold) and VM26 (2.7 fold) but significantly more sensitive to Ara-C (2.3-fold) compared to the intermediate age group. When analyzing infants <1 year of age similar results were found. ProB ALL cells (seven infants <1.5 years; eight children >1.5 years) were significantly more resistant to glucocorticoids, Asp, thiopurines, anthracyclines and Ifos compared to cALL/preB ALL but more sensitive to Ara-C. Cells from children >10 years were significantly more resistant to Pred, DXM, Asp, Ida and 6-MP. T-ALL cells showed a strong resistance to Pred, Asp and VCR and a mild but significant resistance to all other drugs except thiopurines and VM26. We conclude that the poor prognosis of infant ALL is associated with a resistance to glucocorticoids and Asp. However, ALL cells from infants show a relatively high sensitivity to Ara-C which suggests that infants with ALL might benefit from treatment schedules that incorporate more Ara-C than the current treatment protocols. PMID- 9737682 TI - RT-PCR method with increased sensitivity shows persistence of PML-RARA fusion transcripts in patients in long-term remission of APL. AB - RT-PCR methods have been developed, to date, by various groups to amplify the PML RARA fusion gene produced by the t(15;17) in APL patients. However, these methods lack the necessary sensitivity to detect minimal residual disease (MRD) below the level of 1 leukaemic cell in 10(4) cells. Patients who test positive by these methods after treatment are likely to relapse. However, up to 25% of patients who test negative after treatment relapse within a short period. We have developed a 'hot-start' RT-PCR method for the amplification of PML-RARA with increased sensitivity at the level of two leukaemic cells in 10(6) cells. Using this method we were able to detect MRD in seven out of 15 patients tested in remission. Of the 11 patients in medium to long-term remission, five patients tested positive. None of these 11 patients tested positive with the standard RT-PCR. These results show that some patients in remission of APL continue to express PML-RARA even in long-term remission, when they can be considered clinically 'cured' of their disease. PMID- 9737683 TI - Inhibition of erythroid differentiation and induction of megakaryocytic differentiation by thrombopoietin are regulated by two different mechanisms in TPO-dependent UT-7/c-mpl and TF-1/c-mpl cell lines. AB - Thrombopoietin (TPO) regulates megakaryocytic (MK) maturation and platelet production. Molecular and cellular mechanisms of the TPO-induced MK differentiation are not totally understood. In order to develop cellular models to study these mechanisms, we introduced c-mpl into UT-7 and TF-1 cells by means of a retroviral vector and compared the effects of TPO on these two cell lines. UT-7 and TF-1 cell lines are two factor-dependent leukemic cell lines with an erythroid and MK phenotype. They proliferate in response to IL-3, GM-CSF and EPO, but not to TPO. The erythroid differentiation of both cell lines can be markedly increased by EPO. Several UT-7/c-mpl and TF-1/c-mpl cell clones which express different levels of the c-mpl protein (Mpl) were obtained and all became TPO dependent for their proliferation. The UT-7/c-mpl clones, but not the TF-1/c-mpl clones, were capable of undergoing MK differentiation in response to TPO. This was demonstrated by the increase in MK markers (GPIIb, GPIIIa, GPIb alpha, GPIX and vWF), the appearance of cytoplasmic alpha-granules, intracellular membranes resembling demarcation membranes which were immunologically labeled with an GPIIb/IIIa anti-antibody, and a small percentage of polyploid cells (8N and 16N). In contrast, TPO inhibited the erythroid program of differentiation (glycophorin A, beta-globin and EPO receptor) as well as the differentiative activity of EPO in both UT-7/c-mpl and TF-1/c-mpl clones. It is noteworthy that the differentiative effect of EPO in TF-1/c-mpl cells was associated with an increase in GATA-1 transcripts which was totally suppressed by TPO. Overall the effects of TPO are the same as those of phorbol myristate acetate (PMA) which also induces MK differentiation and inhibits erythroid differentiation. These results suggest that: (1) Mpl expression is necessary but not sufficient for induction of MK differentiation; and (2) induction of Mk differentiation and inhibition of erythroid differentiation by TPO involve different signaling pathways; the pathway involved in the inhibition of erythroid differentiation might be related to a downregulation of GATA-1 expression in TF-1 cells. PMID- 9737684 TI - Lung resistance protein (LRP) gene expression in adult acute myeloid leukemia: a critical evaluation by three techniques. AB - The role of LRP in clinical drug resistance in acute myeloid leukemia (AML) is controversial. We therefore compared multiple assays, including RT-PCR, immunocytochemistry (ICC) and flow cytometry (FC), in 10 cell lines and in 47 fresh and thawed AML cells in order to validate and to quantitate measures for LRP phenotype detection. We also compared different ways of expressing the results. Lastly, in cell lines, we analyzed the 50% lethal concentration (LC50), by MTT assay, of cisplatin which could estimate the functionality of LRP. The reproducibility of LRP detection measured by RT-PCR, ICC and FC was good. In the same way, within the same technique, there was good correlation between the different methods of expressing the results of LRP level. Therefore, the discrepancies noted with the three techniques used were neither a problem of reproducibility nor a problem of results expression. On the other hand, there was only a correlation between ICC and FC, and no correlation between RT-PCR and LRP protein detection techniques. Therefore, RT-PCR is probably not the optimal technique for LRP detection. We have shown in 10 cell lines a higher correlation between FC and LC50 of cisplatin than between ICC and LC50 of cisplatin and no correlation between RT-PCR and LC50 of cisplatin. For five patients, there was a dissociation between ICC and FC. Four patients were positive by FC and negative by ICC and only one patient was negative by FC and positive by ICC. Therefore, if in vitro resistance to cisplatin represents the functionality of LRP, we recommend the use of FC rather than ICC to detect LRP expression. Besides the measurement of LRP as a diagnostic tool in the evaluation of resistance to chemotherapy in patients with AML, we urgently need to establish a functional test in order to assess LRP activity. PMID- 9737685 TI - Stem cell factor enhances the adhesion of AML cells to fibronectin and augments fibronectin-mediated anti-apoptotic and proliferative signals. AB - Acute myeloid leukaemia (AML) cells express the SCF receptor c-kit (CD117) on their cell surface and demonstrate enhanced adhesion to fibronectin (FN) following exposure to stem cell factor (SCF). Increased adhesion occurs within 5 min, is dose dependent, and persists beyond 2 h. Baseline and enhanced adhesion occur through the surface FN receptor very late antigen-5 (VLA-5, CD49e/CD29) which is expressed by AML cells. Unstimulated AML cells exposed to FN undergo less apoptosis than controls (inhibition 22.5 +/- 7.0%, P = 0.02, n = 8). Exposure to SCF alone without FN also inhibits AML cell apoptosis (by 19.0 +/- 7.7% compared to controls, P = 0.06, n = 8). Simultaneous exposure to SCF and FN increases the inhibition of AML cell apoptosis to 37.8 +/- 7.9% (P = 0.005 compared to control, P = 0.04 compared to FN alone, P = 0.06 compared to SCF alone) demonstrating that SCF not only enhances the propensity of AML cells to adhere to FN, but also results in an additive survival benefit following FN contact. Some but not all the reduction in apoptosis is mediated through VLA-5. The combination of SCF and FN also affects proliferation, resulting in a synergistic enhancement of AML cell proliferation in half the cases studied. When normal CD34+ human haemopoietic progenitors were studied, FN had little effect on their apoptosis and failed to enhance the anti-apoptotic effect of SCF. It did, however, synergise with SCF in promoting CD34+ cell proliferation. Exposure of AML cells to SCF and FN, both of which can be found in high concentration in the bone marrow stroma, inhibits apoptosis. Cytokines and extracellular matrix proteins augment each others' effects since SCF enhances adhesion to fibronectin, which in turn augments the survival signal delivered by the cytokine alone. Cytokine and adhesion receptors can combine to affect cell characteristics including proliferation and survival. PMID- 9737686 TI - The induction of apoptosis and cell cycle arrest by arsenic trioxide in lymphoid neoplasms. AB - Arsenic trioxide (As2O3) has recently been shown to induce complete remission in acute promyelocytic leukemia (APL). As2O3 reportedly has dose-dependent dual effects on APL cells, triggering apoptosis at relatively high concentrations and inducing differentiation at lower concentrations. However, its effect is still controversial for other AML cells and hematological neoplasms. We studied the in vitro effect of As2O3 on lymphoid lineage cells: lymphoma cell lines, NOL-3, Raji and Daudi, a myeloma cell line, NOP-1, normal peripheral blood lymphocytes (PBL), non-Hodgkin's lymphoma (NHL) cells and chronic lymphocytic leukemia (CLL) cells, and compared it with the effect on APL cell line, NB4, as well as other myeloid cell lines, HL-60 and NKM-1. As2O3 at a concentration of 1 micromol/l markedly inhibited both proliferation and viability of NB4, NOP-1, NOL-3 and NKM-1 cells, but it reduced only viability in normal PBL, CLL cells and NHL cells. As2O3 induced apoptosis and down-regulated bcl-2 expression in NB4, NOP-1 and NKM-1 cells. On the other hand, in HL-60, Raji and Daudi cells, 1 micromol/l As2O3 inhibited only the proliferation weakly, and neither induced apoptosis nor down regulated bcl-2 expression, but arrested only cell cycle at G1 phase. As2O3 at a low concentration of 0.1 micromol/l had no effect on proliferation and viability of these cells except for NB4. These results showed that As2O3 exerted variable and definite effects on lymphoid cells and indicated that As2O3 might be clinically useful in lymphoid neoplasms such as malignant lymphoma and CLL. PMID- 9737687 TI - Vitamin K2 selectively induces apoptosis of blastic cells in myelodysplastic syndrome: flow cytometric detection of apoptotic cells using APO2.7 monoclonal antibody. AB - We have previously reported that vitamin K2 (VK2) but not VK1 has a potent apoptosis-inducing effect on freshly isolated leukemia cells from patients with various types of leukemia. By multi-color flow cytometric analysis using monoclonal antibody (mAb), APO2.7, which detects mitochondrial 7A6 antigen specifically expressed by cells undergoing apoptosis, we further investigated the apoptosis-inducing effect of VK2 on minor populations of leukemic blast cells in bone marrow from patients with myelodysplastic syndrome (MDS) and overt myeloid leukemia (post-MDS AML). Limiting dilution of CD95 (anti-Fas) mAb-treated apoptotic Jurkat cells with nonapoptotic CTB-1 cells revealed that APO2.7 positive Jurkat cells were consistently detectable by flow cytometry when present at levels of at least 5% in the CTB-1 suspension. In patient samples the gating area for leukemic clone was determined using cell surface antigen-specific mAbs conjugated with either fluorescein isothionate (FITC) or phycoerythrin (PE) and subsequently the cells stained with phycoerythrin cyanine (PE-Cy5)-conjugated APO2.7 mAb were assessed within the gating area of the leukemic clone for monitoring apoptosis. Treatment of the bone marrow mononuclear cells with 3-10 microM of VK2 (menaquinone-3, -4 and -5) in vitro potently induced apoptosis of the leukemic blast cells as compared with the untreated control cells in all 15 MDS patients tested. This effect was more prominent on blastic cells than that on mature myeloid cells such as CD34-/CD33+ gated cells. In addition, VK2 performed much less effectively on CD3-positive lymphoid cells. In contrast to VK2, VK1 did not show apoptosis-inducing activity. These data suggest that VK2 may be used for treatment of patients with MDS in blastic transformation. PMID- 9737688 TI - High frequency of fusion transcripts of exon 11 and exon 4/5 in AF-4 gene is observed in cord blood, as well as leukemic cells from infant leukemia patients with t(4;11)(q21;q23). AB - The MLL gene located on chromosome 11q23 and its translocation to the AF-4 gene located on chromosome 4q21 play a pivotal role in leukemogenesis in infancy. Studies of identical leukemic twins have provided evidence of the MLL rearrangement as a fetal event during pregnancy. We analyzed the presence and frequency of the MLL/AF-4 rearrangement in normal cord blood. Although no chimeric mRNA of MLL or AF-4 was detected in 65 cord blood samples, in-frame fusion transcripts of exon 11 and exon 4 or 5 of the AF-4 gene were detected in three of the samples by a nested polymerase chain reaction. When primers of exon 11 and exon 5 of the AF-4 gene were used, two forms of fusion transcripts (AF-4 exon 11/4 or exon 11/5) were detected in 20 of the 65 cord blood samples (31%) and also four of six leukemic cell samples with t(4;11) (67%), whereas such transcripts were not observed in any of 21 peripheral blood samples nor in fetal fibroblasts. These findings suggest that the in-frame fusion of exon 11 and exon 4 or 5 of the AF-4 gene frequently occurs in hematopoietic cells during the intrauterine period, even in a healthy fetus. Although it is unknown whether the proteins of the AF-4 fusion transcripts have some functions, the instability of the AF-4 gene may be associated with the leukemogenesis of infant leukemia. PMID- 9737689 TI - Alternative splicing in wild-type AF10 and CALM cDNAs and in AF10-CALM and CALM AF10 fusion cDNAs produced by the t(10;11)(p13-14;q14-q21) suggests a potential role for truncated AF10 polypeptides. AB - The t(10;11)(p13;q14-21) is a non-random translocation that occurs primarily in T cell acute lymphoblastic leukemias (T-ALL), but has also been observed in leukemias and lymphomas of diverse lineages. In U937, a cell line established from a diffuse histiocytic lymphoma, a t(10;11)(p13;q14-21) fuses AF10 to CALM. AF10 is also fused to MLL by a translocation that appears quite similar at the cytogenetic level, the t(10;11)(p12;q23). Fluorescence in situ hybridization studies have demonstrated that AF10 and CALM are also involved in other hematological malignancies containing t(10;11)(p13;q21), but no data are available concerning the molecular details of AF10-CALM fusion in primary leukemias. Using RT-PCR, we amplified multiple different isoforms of AF10-CALM and CALM-AF10 fusion cDNAs from a primary T cell ALL containing a t(10;11)(p13 14;q14-21). These cDNAs arose via alternative splicing of exons from both AF10 and CALM, which we demonstrated can also occur in the native genes. We identified at least two novel AF10 exons that can be included in wild-type and fusion cDNAs. The majority of the AF10 and AF10-CALM cDNA isoforms that we identified are predicted to encode for truncated AF10 polypeptides, raising the possibility that these might have important cellular functions in normal and malignant cells, perhaps by acting as dominant negative inhibitors of full-length AF10 or related proteins. PMID- 9737690 TI - Molecular characterization of jumping translocations reveals spatial and temporal breakpoint heterogeneity. AB - Jumping translocations (JT) are characterized by the relocalization of the same part of a donor to several recipient chromosomes. Although JT occasionally are constitutional, most are associated with hematologic malignancies. In such cases, JT usually arise during disease progression and are associated with poor prognosis. Despite its clinical importance, this cytogenetic phenomenon has not been characterized at the molecular level. We have analyzed JT in a juvenile chronic myelomonocytic leukemia that subsequently transformed to an acute myeloid leukemia. Detailed fluorescence in situ hybridization (FISH) analyses showed that the cytogenetically identical donor breakpoint at 3q21 was highly heterogeneous. In fact, more than 10 distinct breakpoints, four of which mapped within YACs, were identified. Analyses of samples during disease progression showed that the breakpoint complexity decreased, indicating clonal selection. Hence, the 3q21 breakpoints displayed a spatial as well as a temporal heterogeneity, revealing that JT are highly unstable, showing great variation in the size of donor segment. The breaks at the recipient chromosomes were mapped within the subtelomeric regions. The general telomere length was not affected and an underlying replication error resulting in microsatellite instability was excluded. We conclude that the emergence of JT is unlikely to cause fusion genes or to affect the expression of genes located in the breakpoint regions. The identification of YACs spanning the breakpoints, ie, YACs 913c7, 937g5, 948c2 and 955g1, may facilitate the isolation of DNA sequences leading to a genetic instability associated with the origin of multiple translocations. PMID- 9737691 TI - Different patterns of homozygous p16INK4A and p15INK4B deletions in childhood acute lymphoblastic leukemias containing distinct E2A translocations. AB - The p16INK4A (p16) and p15INK4B (p15) tumor suppressor genes are inactivated by homozygous gene deletion and p15 promoter hypermethylation in a significant proportion of childhood acute lymphoblastic leukemias (ALLs). However, little is known about the potential association between p16/p15 gene alterations and specific genetic abnormalities implicated in leukemogenesis. The t(1;19)(q23;p13) and t(17;19)(q21-22;p13) are non-random translocations observed in childhood ALL that create distinct E2A fusion proteins: E2A-PBX1 and E2A-HLF, respectively. Previously, a negative association was found between the t(1;19) and homozygous p16/p15 deletions. In this study we determined p16 and p15 gene status in additional t(1;19)+ ALLs and compared this incidence to that observed in t(17;19)+ ALLs. No homozygous p16 or p15 deletions were observed among 13 t(1;19)+ ALLs analyzed. In contrast, homozygous deletions of both p16 and p15 were present in two of four t(17;19)+ ALLs. None of 10 t(1;19)+ ALLs contained p15 promoter hypermethylation. In contrast, one of the two t(17;19)+ ALLs that lacked p15/p16 homozygous deletions showed probable hemizygous p15 hypermethylation. We conclude that homozygous p16 and/or p15 deletions and p15 hypermethylation rarely accompany E2A-PBX1 fusion, but occur in concert with E2A HLF fusion in a subset of t(17;19)+ ALLs. These findings suggest that there may be different modes of cooperative leukemogenesis in ALLs associated with different E2A fusion proteins. PMID- 9737692 TI - Truncated c-Myb expression in the human leukemia cell line TK-6. AB - The c-MYB proto-oncogene encodes a transcription factor which plays an important role in hematopoiesis. We detected truncated c-MYB mRNA (2.0 kb) and c-Myb protein (55 kDa) in the TK-6 cell line, which was established from a patient with chronic myelogenous leukemia in T cell blast crisis. Mutated c-MYB cDNA clone (WTK-1) was isolated from a TK-6 cell cDNA library and sequenced in its entirety. Compared with the wild-type human c-MYB sequence, the WTK-1 sequence diverged at the 3' ends of exons 9. A termination codon was present as the second codon downstream from the point of divergence and was followed by a previously unknown rearranged sequence. The conceptual protein encoded by WTK-1 (Myb(TK-6)) comprises 402 amino acids and lacks the negative regulatory domain of the normal c-Myb, reminiscent of the activated form of Myb protein. Luciferase reporter assay in NIH3T3 cells showed that the expression vector encoding Myb(TK-6) stimulated Myb-regulated mim-1 promoter more effectively than that encoding wild type human c-Myb, suggesting that Myb(TK-6) is functional as a transcription factor, and thus as a potential transforming protein. Southern blot and mutant allele-specific polymerase chain reaction analyses showed that the same rearrangement of the c-MYB gene in TK-6 was present in late, but not in early, specimens obtained from the patient, indicating that this mutation had been acquired during disease progression. PMID- 9737693 TI - Models of lineage switching in hematopoietic development: a new myeloid-committed eosinophil cell line (YJ) demonstrates trilineage potential. AB - A new human leukemia cell line with an eosinophilic phenotype, designated YJ, was established from the peripheral blood cells of a patient with chronic myelomonocytic leukemia (CMMoL) with eosinophilia. When cultured in RPMI 1640 medium containing 10% fetal bovine serum, most YJ cells were myeloblastoid with a small number of the cells having eosinophilic granules. Cell surface markers in the YJ cells were positive for CD33 and were negative for CD34, CD16 and CD23. The eosinophilic characteristics of YJ cells were confirmed by histochemical staining with Fast-Green/Neutral-Red and by the expression of mRNAs for eosinophil-associated granule proteins, eosinophil cationic protein (ECP), eosinophil-derived neurotoxin (EDN), eosinophil peroxidase (EPO), and major basic protein (MBP), and for the Charcot-Leyden crystal (CLC) protein. The YJ cells could be induced towards monocytic differentiation by stimulation with phorbol 12 myristate 13-acetate (PMA). The monocytic characteristics of YJ cells treated with PMA were confirmed by morphological analysis with alpha-naphthyl butyrate esterase staining, by CD14 expression, and by increased expression of Egr-1 mRNA. Furthermore, YJ cells could be differentiated towards the neutrophil lineage by stimulation with all-trans retinoic acid (RA). YJ cells treated in vitro with 2 microM RA differentiated into metamyelocytes and band neutrophils, and increased the number of nitroblue tetrazolium (NBT)-positive cells and increased gp91phox mRNA expression. Thus, the YJ cell line exhibited eosinophilic characteristics, but was able to differentiate to the monocytic or neutrophilic lineages in response to PMA or RA, respectively. The expression of genes for transcription factors involved in myeloid differentiation was evaluated by Northern blot analysis. Increased expression of Egr-1 was observed with macrophage differentiation. In contrast, increased expressions of C/EBPbeta and MZF-1 mRNA occurred with neutrophilic differentiation. The YJ cell line should be useful for elucidating the molecular mechanisms governing lineage switching from the eosinophil to monocytic or neutrophil lineages. PMID- 9737694 TI - Influence of CD34 cell selection on the incidence of mixed chimaerism and minimal residual disease after allogeneic unrelated donor transplantation. AB - Marrow transplantation from unrelated donors has been linked with an increased risk of graft-versus-host disease (GVHD). In an attempt to lower the risk of acute GVHD we used CD34 marrow cell selection for T cell depletion. Since T cell depletion has been linked to an increased risk of relapse and an increased risk of marrow failure, we used PCR amplification of minisatellite sequences to investigate donor cell engraftment and RT-PCR amplification of recurrent chromosomal translocations to investigate the residual disease post-transplant. Twenty-three patients who underwent BMT after positive selection of the CD34 positive cell population were studied. Results were then compared with those of 37 patients who underwent transplantation with unmanipulated marrow graft. Among the 23 patients who received CD34+ selected cell grafts, seven (30%) had evidence of full donor engraftment, 14 had evidence of residual recipient cells (61%), one had a non-take, and one autologous bone marrow recovery. Analysis of the chimaerism status post-transplant in 36 patients who received unmanipulated marrow grafts showed that 31 patients (86%) had evidence of full donor engraftment. The difference in the incidence of mixed chimaerism profile between patients who received unmanipulated marrow graft and those receiving CD34+ selected cell grafts was statistically significant (P< 0.01). Nine patients who received CD34+ selected cell grafts could be analysed for the presence of minimal residual disease post-transplant (one with t(9;22) acute lymphoblastic leukaemia and eight with CML). In the patient transplanted for a Ph-positive acute leukaemia, and in two out of the eight patients with CML, the search fora fusion transcript was consistently negative after transplantation. Among the six patients with evidence of residual disease, three patients also had a mixed chimaerism profile and were given donor lymphocyte infusions. Minimal residual disease study was performed post-transplant in 16 patients who received unmanipulated marrow grafts. In 10 of 14 patients with CML, and in two patients with acute leukaemia the search for a fusion transcript was consistently negative after transplantation. The difference in the incidence of minimal residual disease between patients who received an unmanipulated marrow graft and those receiving CD34+ selected cell grafts was not statistically significantly significant, but numbers of patients included in this analysis are still few. In conclusion, our study highlights the strong influence of graft manipulation on the incidence of mixed chimaerism after transplantation from an unrelated donor. PMID- 9737695 TI - Factors affecting both peripheral blood progenitor cell mobilization and hematopoietic recovery following autologous blood progenitor cell transplantation in multiple myeloma patients: a monocentric study. AB - The aim of the study was to analyze the factors influencing peripheral blood progenitor cell (PBPC) collection after high-dose cyclophosphamide (HDCYC) (7 g/m2) and hematopoietic recovery after autologous transplantation of HDCYC mobilized PBPC (ABPCT) in 116 patients with aggressive multiple myeloma (MM). Following HDCYC 74 patients received hematopoietic growth factors (HGF), either G CSF (n = 19) or GM-CSF (n = 55). All the patients were subsequently planned to undergo ABPCT. PBPC collection was possible for 106 patients. The most important prognostic factor for collection of more than 25 x 10(4) CFU-GM cells/kg and 2 x 10(6) CD34+ cells/kg was the use of HGF (P = 0.002 and 0.009, respectively). Previous use of an alkylating agent, response to treatment before HDCYC, and interval between diagnosis and HDCYC were also significant factors (P = 0.004, 0.025 and 0.001, respectively). The number of CFU-GM cells infused was the most important parameter for rapid and complete hematological recovery after ABPCT (P < 0.0001). Thus the use of HGF post-HDCYC is the major factor which, associated with reduced time between diagnosis and HDCYC and the use of an alkylating agent, could increase the numbers of hematopoietic progenitors collected, and subsequently improve hematopoietic recovery following ABPCT in MM patients. PMID- 9737696 TI - Biweekly THP-COPBLM (pirarubicin, cyclophosphamide, vincristine, prednisone, bleomycin and procarbazine) regimen combined with granulocyte colony-stimulating factor (G-CSF) for intermediate- and high-grade non-Hodgkins's lymphoma. AB - Biweekly THP-COPBLM including pirarubicin (THP), which is thought to be less toxic than doxorubicin, was used to treat non-Hodgkin's lymphoma (NHL) and the remission rate and adverse events were studied in 42 patients younger than 69 years. Complete remission (CR) was achieved in 37 patients (88.1%) and partial remission in five (11.9%). Classified by international prognostic index, CR was achieved in 16 out of 17 low-intermediate-risk patients, 14 out of 16 high intermediate-risk patients and seven out of nine high-risk patients. The 3-year survival rate was 72.1% and the 3-year event-free survival rate was 58%. Grade 3 or higher adverse events included granulocytopenia in 39 patients (92.9%) and thrombocytopenia in seven (16.7%). The biweekly THP-COPBLM regimen appears useful for the treatment of aggressive intermediate- and high-grade NHL, and G-CSF made it possible to shorten the interval between courses of chemotherapy. Further studies regarding adverse events on organs, other than on bone marrow are required to improve the long-term results of combination therapy on NHL. PMID- 9737697 TI - Schedule and concentration-dependent induction of apoptosis in leukemia cells by nitric oxide. AB - Nitric oxide (NO) has potent antiproliferative properties. In previous work we have shown that NO inhibits growth, induces differentiation and modulates gene expression in acute nonlymphocytic leukemia (ANLL) cells. The goal of this work was to determine whether the rate of NO delivery affected its growth inhibition of ANLL cells. We also wanted to determine whether the NO inhibition of ANLL cell growth is associated with the induction of apoptosis. We treated HL-60 and U937 cells with three compounds that generate the same amount of NO but at different rates. MAMA-NO, PAPA-NO and DETA-NO have half-lives of NO delivery of 2 and 30 min, and 20 h, respectively. The compound with the longest t(1/2) of NO delivery (DETA-NO) was the most potent inhibitor of leukemia cell and colony growth. Furthermore, the NO-induced growth inhibition was associated with apoptosis in a rate and concentration-dependent fashion. PMID- 9737698 TI - N-terminus cleavage of bcl-2 by a novel cellular non-ICE cysteine proteinase. AB - The bcl-2 protein plays an essential role in preventing cell death. Its activity is regulated through association with bcl-2 homologous and nonhomologous proteins and also by serine phosphorylation. We now report that bcl-2 can be proteolytically cleaved towards its N-terminus by a cysteine proteinase present in RL-7 lymphoma cell lysates, yielding a major product of apparent MW 20 kDa, different from the products of bcl-2 cleavage by HIV protease. Moreover, bcl-2 proteins mutated for Asp residues at positions 31 and 34 were efficiently cleaved by RL-7 cell lysates, indicating that this proteolytic activity is distinct from the caspase-3 that cleaves bcl-2 at Asp 34. This bcl-2 cleaving activity is inhibited by E-64 and is therefore distinct from the proteinases of the ICE/Ced-3 family (caspases), whereas reciprocally, ICE (caspase-1) is unable to cleave bcl 2. It is optimally active at pH 5, a feature distinguishing it from calpain, another non-ICE cysteine proteinase which has been associated with apoptosis. This novel bcl-2 cleaving protease, although constitutively present in RL-7 cells and resting peripheral blood lymphocytes (PBL) was upregulated following induction of apoptosis in RL-7 cells or mitogen activation in PBL. The N-terminus of bcl-2 which contains the BH4 domain that binds the kinase Raf-1 and the phosphatase calcineurin is essential for anti-apoptotic activity. Its cleavage might provide a novel post-translational mechanism for regulating bcl-2 function and could amplify ongoing programmed cell death. PMID- 9737699 TI - High rate of chromosome abnormalities detected by fluorescence in situ hybridization using BCR and ABL probes in adult acute lymphoblastic leukemia. AB - The value of dual-color fluorescence in situ hybridization (FISH) with BCR and ABL probes for the detection of the Philadelphia (Ph) translocation and of other alterations involving ABL and/or BCR was evaluated in adult acute lymphoblastic leukemia (ALL). One hundred and four patients were studied prospectively using interphase nuclei FISH, chromosome analysis (CA), and PCR assays for the chimeric BRC/ABL transcript. FISH detected a Ph translocation in 24 cases (23.1%), as was confirmed by CA and/or PCR. FISH revealed a false positive diagnosis of a Ph translocation in four cases (5% false positive rate). Among 54 cases with combined FISH, CA and PCR assays, FISH failed to establish a correct diagnosis in 3.7%, PCR in 5.6%, and CA in 7.4%. The combination of two screening methods led to discrepant results in 9.3% (FISH + PCR), 11.1% (FISH + CA), or 13% (CA + PCR) of the cases. In seven of 80 (8.8%) Ph-negative patients, gain of BCR and/or ABL was identified. Overall, FISH detected alterations of the BCR and/or ABL genes with an incidence of 29.8% of the current study. Due to the possibility of false positive diagnosis of a Ph translocation using dual-color FISH the combination with chromosome and/or RT-PCR analyses is recommended in adult ALL patients. PMID- 9737700 TI - Detection of minimal residual disease in patients with AML1/ETO-associated acute myeloid leukemia using a novel quantitative reverse transcription polymerase chain reaction assay. AB - The AML1/ETO fusion transcript can be detected by reverse transcription polymerase chain reaction (RT-PCR) in patients with t(8;21)-associated acute myeloid leukemia (AML) in long-term complete remission (CR). Quantitation of the amount of the fusion transcript during CR may therefore be more predictive of cure or relapse than a simple qualitative assessment. Real Time PCR, a fluorometric-based technique, allows simple and rapid quantitation of a target sequence during the extension phase of PCR amplification, in contrast to end point quantitative methods. Six patients with t(8;21)(q22;q22) AML, who achieved CR were studied by Real Time RT-PCR at different time intervals following diagnosis and high-dose cytarabine and anthracycline-based induction therapy. Five patients had a diagnostic bone marrow (BM) sample available for molecular analysis. Each patient showed > or = 10(3) copies of the AML1/ETO fusion transcript at diagnosis, and each showed a 2- to 4-log decrease in copy number following successful induction chemotherapy. This is comparable to the log-fold reduction in leukemic blasts that is thought to occur in patients successfully cytoreduced into CR by induction chemotherapy. The sixth patient showed a relatively high copy number immediately following successful remission induction chemotherapy, which continued to increase during early CR and was later followed by relapse. Real Time RT-PCR appears to offer advantages over previously used quantitative RT-PCR methods by providing absolute quantitation of the target sequence, expanding the dynamic range of quantitation to over six orders of magnitude, eliminating the post-PCR processing, and reducing labor and carryover contamination. These features make this an attractive method to prospectively evaluate the prognostic value of AML1/ETO fusion transcript quantitation in a larger patient population with t(8;21)(q22;q22) AML in CR. PMID- 9737701 TI - A case of cryptic acute promyelocytic leukemia. PMID- 9737702 TI - PML/RAR alpha APL with undifferentiated morphology and stem cell immunophenotype. PMID- 9737703 TI - Response from Estey to X Thomas et al. PMID- 9737704 TI - Ongoing somatic mutations of the immunoglobulin gene in MALT lymphoma with widespread MLP-type polypoid lesions. PMID- 9737706 TI - Identification of a BCR polymorphism in a bone marrow donor: diagnostic implications. PMID- 9737705 TI - Clonal myelodysplastic cells present in apheresis product before transplantation. PMID- 9737707 TI - Early detection of AML1/MTG8 fusion mRNA by RT-PCR in the bone marrow cells from a patient with isolated granulocytic sarcoma. PMID- 9737708 TI - Influence of increased body mass index on drug toxicity in patients with acute promyelocytic leukemia. PMID- 9737709 TI - From molecular analysis to medical practice: recent studies reveal new therapeutic targets for an old medicine. PMID- 9737710 TI - Cyclooxygenase in biology and disease. AB - Cyclooxygenase (COX), the key enzyme required for the conversion of arachidonic acid to prostaglandins was first identified over 20 years ago. Drugs, like aspirin, that inhibit cyclooxygenase activity have been available to the public for about 100 years. In the past decade, however, more progress has been made in understanding the role of cyclooxygenase enzymes in biology and disease than at any other time in history. Two cyclooxygenase isoforms have been identified and are referred to as COX-1 and COX-2. Under many circumstances the COX-1 enzyme is produced constitutively (i.e., gastric mucosa) whereas COX-2 is inducible (i.e., sites of inflammation). Here, we summarize the current understanding of the role of cyclooxygenase-1 and -2 in different physiological situations and disease processes ranging from inflammation to cancer. We have attempted to include all of the most relevant material in the field, but due to the rapid progress in this area of research we apologize that certain recent findings may have been left out. PMID- 9737711 TI - Matrix metalloproteinases: structures, evolution, and diversification. AB - A comprehensive sequence alignment of 64 members of the family of matrix metalloproteinases (MMPs) for the entire sequences, and subsequently the catalytic and the hemopexin-like domains, have been performed. The 64 MMPs were selected from plants, invertebrates, and vertebrates. The analyses disclosed that as many as 23 distinct subfamilies of these proteins are known to exist. Information from the sequence alignments was correlated with structures, both crystallographic as well as computational, of the catalytic domains for the 23 representative members of the MMP family. A survey of the metal binding sites and two loops containing variable sequences of amino acids, which are important for substrate interactions, are discussed. The collective data support the proposal that the assembly of the domains into multidomain enzymes was likely to be an early evolutionary event. This was followed by diversification, perhaps in parallel among the MMPs, in a subsequent evolutionary time scale. Analysis indicates that a retrograde structure simplification may have accounted for the evolution of MMPs with simple domain constituents, such as matrilysin, from the larger and more elaborate enzymes. PMID- 9737712 TI - Alzheimer's amyloid beta and lipid metabolism: a missing link? PMID- 9737713 TI - BRE: a modulator of TNF-alpha action. AB - A stress-responsive gene highly expressed in brain and reproductive organs (BRE) is down-regulated after UV irradiation, DNA damaging agents, or retinoic acid treatment. The human BRE gene encodes a mRNA of 1.9 kb, which gives rise to a protein of 383 amino acids with a molecular size of 44 kilodaltons. BRE is not homologous to any known gene and its function has not been defined. Here we report that BRE was identified multiple times in a yeast two-hybrid screen of a murine cerebellar cDNA library, using the juxtamembrane domain of the p55 tumor necrosis factor alpha (TNF) receptor. The interaction between the p55 receptor and BRE was verified by an in vitro biochemical assay by using recombinant fusion proteins and by co-immunoprecipitation of transfected mammalian cells. In the yeast two-hybrid assay, BRE specifically interacted with p55 TNF receptor but not with other TNF family members such as the Fas receptor, the p75 TNF receptor, and p75 neurotrophin receptor. Overexpression of BRE inhibited TNF-induced NFkappaB activation, indicating that the interaction of BRE protein with the cytoplasmic region of p55 TNF receptor may modulate signal transduction by TNF-alpha. PMID- 9737714 TI - Expression and activity of prostaglandin endoperoxide synthase-2 in agonist activated human neutrophils. AB - Proinflammatory agents were assessed for their capacity to stimulate the expression of the inducible cyclooxygenase isoform (COX-2) in human neutrophils. A number of agents, including PMA, opsonized bacteria and zymosan, LPS, GM-CSF, TNF-alpha, and fMLP, induced COX-2 protein expression through signaling pathways involving transcription and protein synthesis events. Northern blots showed that freshly isolated neutrophils expressed low levels of COX-2 mRNA, which rapidly increased after incubation with inflammatory agents. A characterization of the signal transduction pathways leading to COX-2 protein expression was initiated. In LPS-treated neutrophils, efficient induction of COX-2 required the presence of serum and involved ligand binding to the CD14 surface antigen. The specific inhibitor of p38 mitogen-activated protein kinase (p38 MAPK), SB 203580, had little effect on the induction of COX-2 expression in neutrophils, in contrast to what had been previously observed with other inflammatory cell types. Depending on the agonist present, ethanol differentially blocked the stimulated expression of COX-2, raising the possibility that phospholipase D activation might take part in the process of COX-2 induction. Major COX-2-derived prostanoids synthesized by inflammatory neutrophils were identified by liquid-chromatography and tandem mass spectrometry as TXA2 and PGE2. The agonist-induced synthesis of TXA2 and PGE2 was effectively blocked by cycloheximide and by the specific COX-2 inhibitor NS-398. These results show that COX-2 can be induced in an active state by different classes of inflammatory mediators in the neutrophil. They support the concept that, in these cells, the COX-2 isoform is preeminent over COX-1 for the stimulated-production of prostanoids, and also suggest that neutrophil COX-2 displays a distinct profile of expression among circulatory cells. PMID- 9737715 TI - Altered regulation of L-type channels by protein kinase C and protein tyrosine kinases as a pathophysiologic effect in retinal degeneration. AB - The effect of protein tyrosine kinases (PTK) on L-type calcium channels in cultured retinal pigmented epithelium (RPE) from rats with retinal dystrophy was investigated. Barium currents through Bay K 8644 (10(-6) M) sensitive L-type channels were measured using the patch-clamp technique. The current density of L type currents is twice as high and the inactivation time constants are much slower than in cells from nondystrophic control rats. Application of the PTK blockers genistein, lavendustin A, and herbimycin A (all 5 x 10(-6) M) led to an increase of L-type currents. Intracellular application of pp60c-src (30 U/ml) via the patch pipette led to a transient decrease of L-type currents. The protein kinase A (PKA) and PKG blocker H9 (10(-6) M) showed no effect on L-type currents. However, the protein kinase C blocker chelerythrine (10(-5) M) reduced these currents. Up-regulation of PKC by 10(-6) M 4beta-phorbol-12 myristate-13 acetate (PMA) led to a decrease of L-type currents. Additional application of genistein led to a further decrease of these currents. However, intracellular application of pp60(c-src) in PMA-treated cells led to a transient increase of L-type currents. Investigating the calcium response to bFGF application showed that RPE cells from RCS rats used different pathways than control RPE cells to increase cytosolic free calcium. This different pathway does not involve the activation of L-type channels. The present study with RPE cells from rats with retinal dystrophy shows a changed integration of PTK and PKC in channel regulation. Considering the altered response to bFGF in RCS-RPE cells, this disturbed regulation of L-type channels by tyrosine kinases is involved in the etiology of retinal degeneration in RCS rats. PMID- 9737716 TI - Activation of PDGF receptor alpha in vascular smooth muscle cells by mechanical stress. AB - Hypertension increases mechanical force on the arterial wall by as much as 30%, resulting in marked alterations in signal transductions and gene expression in vascular smooth muscle cells (VSMCs) that contribute to matrix protein synthesis, cell proliferation, and differentiation. How the mechanical stimuli are converted into a biological signal in cells has yet to be studied. We investigated the role of both cyclic strain and shear stresses in initiating the cellular signaling on cultured VSMCs and found that mechanical forces evoked activation of mitogen activated protein kinases, followed by enhanced DNA binding activity of transcription factor AP-1. Physical forces rapidly induced phosphorylation of platelet-derived growth factor receptor (PDGFR) alpha, an activated state. When GRB2, an adapter protein, was immunoprecipitated from treated VSMCs followed by Western blot analysis with anti-phosphotyrosine, -PDGFR alpha, and -GRB2 antibodies, respectively, phosphotyrosine positive staining was observed on PDGFR alpha bands of the same blot in stretch-stressed VSMCs, supporting the mechanical stress-induced activation of PDGFR alpha. Conditioned medium from stretch stressed VSMCs did not result in PDGFR alpha phosphorylation, and antibodies binding to all forms of PDGFs did not block stress-induced PDGFR alpha activation. Thus, mechanical stresses may directly perturb the cell surface or alter receptor conformation, thereby initiating signaling pathways normally used by growth factors. PMID- 9737717 TI - Glutathione peroxidase protects mice from viral-induced myocarditis. AB - Glutathione peroxidase 1 (GPX-1) is a selenium-dependent enzyme with antioxidant properties. Previous investigations determined that mice deficient in selenium developed myocarditis when infected with a benign strain of coxsackievirus B3 (CVB3/0). To determine whether this effect was mediated by GPX-1, mice with a disrupted Gpx1 gene (Gpx1-/-) were infected with CVB3/0. Gpx1-/- mice developed myocarditis after CVB3/0 infection, whereas infected wild-type mice (Gpx1+/+) were resistant. Sequencing of viruses recovered from Gpx1(-/-)-infected mice demonstrated seven nucleotide changes in the viral genome, of which three occurred at the G residue, the most easily oxidized base. No changes were found in virus isolated from Gpx1+/+ mice. These results demonstrate that GPX-1 provides protection against viral-induced damage in vivo due to mutations in the viral genome of a benign virus. PMID- 9737718 TI - Inhibition of TGF-beta-stimulated CTGF gene expression and anchorage-independent growth by cAMP identifies a CTGF-dependent restriction point in the cell cycle. AB - CTGF is a 38 kDa cysteine-rich peptide whose synthesis and secretion are selectively induced by transforming growth factor beta (TGF-beta) in connective tissue cells. We have investigated the signaling pathways controlling the TGF beta induction of connective tissue growth factor (CTGF) gene expression. Our studies indicate that inhibitors of tyrosine kinases and protein kinase C do not block the signaling pathway used by TGF-beta to induce CTGF gene expression. In contrast, elevation of cAMP levels within the target cells by a variety of methods blocked the induction of CTGF by TGF-beta. Furthermore, agents that elevate cAMP blocked the induction of anchorage-independent growth (AIG) by TGF beta. Inhibition of AIG could be overcome by the addition of CTGF, indicating that it was not a general inhibition of growth but a selective inhibition of CTGF synthesis that is responsible for the inhibition of TGF-beta-induced AIG by cAMP. Kinetic studies of the induction of DNA synthesis by CTGF in cells arrested by cAMP indicate that the block occurs in very late G1. These and other studies in monolayer cultures suggest that the CTGF restriction point in the cell cycle is distinct from the adhesion-dependent arrest point. PMID- 9737719 TI - Factor XIIIa as a nerve-associated transglutaminase. AB - Recent findings have led to changes in the traditional concept of nerve recovery, including the realization that injured nerves, like any other injured tissue, need the assistance of blood-derived cells and factors in order to heal. We show that factor XIIIa (FXIIIa, the potentially active a2subunit of factor XIII), an enzyme that participates in blood coagulation by stabilizing the fibrin clot, is also active in the nervous system where it may play a key role in the healing of injured tissue. We demonstrate that the plasma, macrophages and nerves of fish contain a 55 kDa form of transglutaminase that cross-reacts immunologically with the a-subunit of FXIII in mammals (80 kDa). The fish enzyme in the plasma, unlike its mammalian counterpart, is active, pointing to a difference in control of the coagulation pathway in the two species. Analysis of FXIIIa expression in mammalian neural tissues and their response to injury revealed high levels of the enzyme in media conditioned by peripheral nerves as compared with medium conditioned by nerves of the central nervous system. Furthermore, similarity was observed in the postinjury behavior of FXIIIa in regenerating nerve tissues (peripheral nervous system of mammals and the central nervous system of fish). We suggest that the postinjury level of factor XIIIa in the nervous system may be related to the tissue's regenerative capacity, and that FXIIIa may therefore be a link underlying a possible association between the processes of blood coagulation and nerve healing. PMID- 9737720 TI - Glucose and glucoincretin peptides synergize to induce c-fos, c-jun, junB, zif 268, and nur-77 gene expression in pancreatic beta(INS-1) cells. AB - To link glucose signaling to its long-term pleiotropic effects in the pancreatic beta-cell, we have investigated whether glucose regulates immediate-early response genes (IEGs) coding for transcription factors implicated in cell proliferation and differentiation. Glucose causes a coordinated transcriptional activation of the IEGs c-fos, c-jun, JunB, zif-268, and nur-77 in the pancreatic beta-cell line INS-1. This activation is entirely dependent on the presence of the cell-permeant cAMP analog chlorophenylthio-cAMP, which has only a modest effect by itself. The accumulation of c-fos, JunB, and nur-77 mRNA occurs at physiological concentrations of glucose (3 to 11 mM), requires a 1-2 h period, and is mimicked by other nutrient stimuli including mannose, leucine plus glutamine, and pyruvate. Glucose is synergistic with the glucoincretin peptides GLP-1 and PACAP-38, whereas these neurohormonal agents have no effect at low (3 mM) glucose. Mechanistically, the synergy between glucose and the glucoincretins is not based on cAMP alone as glucose does not further increase intracellular cAMP in response to GLP-1 and PACAP-38. A role for Ca2+ signaling is inferred, since the L-type Ca2+ channel blocker nifedipine markedly reduces the induction of c-fos and nur-77 by glucose and GLP-1. The induction of IEGs by glucose and chlorophenylthio-cAMP or GLP-1 and the inhibitory effect of nifedipine are also observed in the betaHC9 cell line. The results indicate that GLP-1 and PACAP-38 act as competence factors for the action of glucose on c-fos, JunB, and nur-77. It is suggested that the synergistic effect of glucose and glucoincretins on IEG expression plays an important role in the adaptive processes of the beta-cell to hyperglycemia. PMID- 9737721 TI - Age-associated decline in ascorbic acid concentration, recycling, and biosynthesis in rat hepatocytes--reversal with (R)-alpha-lipoic acid supplementation. AB - Ascorbic acid recycling from dehydroascorbic acid and biosynthesis from gulono 1,4-lactone were used as measures of cellular response capacity to increased oxidative stress induced by tert-butylhydroperoxide. The hepatic ascorbic acid concentration was 54% lower in cells from old rats when compared to cells isolated from young rats (P<0.0005). Freshly isolated hepatocytes from old rats exhibited a significantly decreased ascorbic acid recycling capacity in response to oxidative stress (P<0.005) compared to cells from young rats. Ascorbic acid synthesis in these cells from old animals was unaffected by various concentrations of tert-butylhydroperoxide, but amounted to only approximately half of the biosynthetic rate when compared to cells from young animals (P<0.001). Cells from young animals were not significantly affected by the tert butylhydroperoxide treatments. The results demonstrate a declining ability with age to respond to increased oxidative stress. (R)-alpha-Lipoic acid, a mitochondrial coenzyme, is a powerful antioxidant. A two-week dietary supplementation of old animals with 0.5% (R)-alpha-lipoic acid prior to cell isolation almost completely reversed the age-associated effects on ascorbic acid concentration (P<0.0001), recycling (P<0.05) and biosynthesis after oxidative stress. These results provide further evidence for the potential of alpha-lipoic acid in treatment of diseases related to oxidative stress. Furthermore, the study extends the value of ascorbic acid as a biomarker of oxidative stress. PMID- 9737722 TI - Identification and expression of a novel isoform of cAMP response element modulator in the human heart. AB - In end-stage human heart failure, excessive beta-adrenergic stimulation of the cAMP-dependent signaling pathway due to enhanced endogenous catecholamines is hypothesized to contribute to expressional alterations of myocardial regulatory proteins. The cAMP response element modulator (CREM) regulates the transcription of cAMP-responsive genes and might be involved in the regulation of cardiac gene expression. Using the reverse transcription polymerase chain reaction, we identified a novel CREM mRNA, CREM-Ib deltaC-X, in the human heart. Overexpression of CREM-Ib deltaC-X decreased cAMP response element (CRE) mediated gene transcription in HIT-T15 cells, and this activity was assigned to the part of the sequence encoding putative internally translated proteins. Two of three possible internally translated proteins were immunologically identified in cells overexpressing CREM-Ib deltaC-X tagged with the hemagglutinin epitope of the influenza virus. Both proteins were expressed in bacteria and showed CRE specific DNA binding, formation of heterodimers with the cAMP response element binding protein (CREB), and inhibition of CREB's binding to the CRE. CREM expression was detected on the mRNA and protein levels in the human heart. We conclude that CREM-Ib deltaC-X generates internally translated repressors of CRE mediated gene transcription, suggesting the first example for the existence and function of human cardiac CREM. PMID- 9737723 TI - A direct protein-protein interaction is involved in the cooperation between thyroid hormone and retinoic acid receptors and the transcription factor GHF-1. AB - The nuclear receptors for thyroid hormone (TRs) and retinoic acid (RARs and RXRs) cooperate with the pituitary-specific transcription factor GHF-1 to activate the rat growth hormone (GH) gene. The GH promoter contains a hormone response element (HRE), which binds TR/RXR and RAR/RXR heterodimers, located close to two binding sites for GHF-1. GHF-1 inhibits binding of TR/RXR and RAR/RXR heterodimers to an isolated HRE. Similarly, the receptors inhibit binding of GHF-1 to its cognate site. These results suggest the existence of direct protein to protein interactions between the receptors and the pituitary transcription factor. This was confirmed by in vitro binding studies with GST fusion proteins, which demonstrated a strong association of GHF-1 with RXR and a weaker interaction with RAR and TR. GHF-1 and the receptor heterodimers form a ternary complex with a fragment of the rat GH promoter, which contains binding sites for both, and GHF-1 increases receptor binding to the promoter when present in limiting conditions. These results suggest that the synergistic activation of the rat GH gene involves protein-DNA interactions as well as a physical association between the nuclear receptors and the pituitary-specific transcription factor GHF-1. PMID- 9737724 TI - Selective MT2 melatonin receptor antagonists block melatonin-mediated phase advances of circadian rhythms. AB - This study demonstrates the involvement of the MT2 (Mel1b) melatonin receptor in mediating phase advances of circadian activity rhythms by melatonin. In situ hybridization histochemistry with digoxigenin-labeled oligonucleotide probes revealed for the first time the expression of mt1 and MT2 melatonin receptor mRNA within the suprachiasmatic nucleus of the C3H/HeN mouse. Melatonin (0.9 to 30 microg/mouse, s.c.) administration during 3 days at the end of the subjective day (CT 10) to C3H/HeN mice kept in constant dark phase advanced circadian rhythms of wheel running activity in a dose-dependent manner [EC50=0.72 microg/mouse; 0.98+/ 0.08 h (n=15) maximal advance at 9 microg/mouse]. Neither the selective MT2 melatonin receptor antagonists 4P-ADOT and 4P-PDOT (90 microg/mouse, s.c.) nor luzindole (300 microg/mouse, s.c.), which shows 25-fold higher affinity for the MT2 than the mt1 subtype, affected the phase of circadian activity rhythms when given alone at CT 10. All three antagonists, however, shifted to the right the dose-response curve to melatonin, as they significantly reduced the phase shifting effects of 0.9 and 3 microg melatonin. This is the first study to demonstrate that melatonin phase advances circadian rhythms by activation of a membrane-bound melatonin receptor and strongly suggests that this effect is mediated through the MT2 melatonin receptor subtype within the circadian timing system. We conclude that the MT2 melatonin receptor subtype is a novel therapeutic target for the development of subtype-selective analogs for the treatment of circadian sleep and mood-related disorders. PMID- 9737725 TI - Sustained hyperglycemia in vitro down-regulates the GLUT1 glucose transport system of cultured human term placental trophoblast: a mechanism to protect fetal development? AB - The trophoblast of human placenta is directly exposed to the maternal circulation. It forms the main barrier to maternal-fetal glucose transport. The present study investigated the effect of sustained hyperglycemia in vitro on the glucose transport system of these cells. Trophoblasts isolated from term placentas and immunopurified were cultured for 24, 48, and 96 h in DMEM containing either 5.5 (normoglycemia) or 25 mmol/l D-glucose (hyperglycemia), respectively. Initial uptake of glucose was measured using 3-O-[14C]methyl-D glucose. Kinetic parameters were calculated as K(M) = 73 mmol/l and Vmax = 29 fmol s(-1) per trophoblast cell. Uptake rates of cells cultured under hyperglycemic conditions did not differ at exogenous D-glucose concentrations in the physiological range (1, 5.5, 10, and 15 mmol/l), but were significantly decreased by 25% (P<0.05) at diabetes-like concentrations (20 and 25 mmol/l) as compared to normoglycemic conditions. This effect was due to a decrease in Vmax ( 50%), whereas K(M) remained virtually unaffected. GLUT1 mRNA levels were lower by 50% (P<0.05; Northern blotting) and GLUT1 protein was reduced by 16% (P<0.05; Western blotting) in trophoblast cells cultured under hyperglycemic vs. normoglycemic conditions. We conclude that prolonged hyperglycemia in vitro reduces trophoblast glucose uptake at substrate concentrations corresponding to blood levels of poorly controlled diabetic gravidas. This effect is due to diminished GLUT1 mRNA and protein expression in the trophoblast. PMID- 9737726 TI - Role of cyclin E and cyclin E-dependent kinase in mitogenic stimulation by cementum-derived growth factor in human fibroblasts. AB - Cementum-derived growth factor (CGF) is a 14 kDa polypeptide sequestered in tooth cementum. It is an IGF-I like molecule that is weakly mitogenic to fibroblasts, but its mitogenic action is synergistically potentiated in the presence of epidermal growth factor (EGF) or serum. We have examined whether the CGF affects cyclin E levels and the activity of cyclin-dependent kinase (Cdk) associated with this cyclin, and whether these changes contribute to the synergism in mitogenic activity between CGF and EGF. Optimal DNA synthesis by serum-starved human gingival fibroblasts required the presence of CGF for 0-12 h and EGF for 0-3 h. Therefore, cells were serum starved for 48 h and then exposed to CGF, EGF, or CGF + EGF. Cells incubated with 10% fetal bovine serum (FBS) served as positive controls. At various time points after the addition of growth factors, cyclin E levels were examined by Western analysis. Cdk associated with cyclin E was immunoprecipitated with anti-cyclin E antibody and kinase activity was measured using H1 histone as substrate. Cyclin E and the H1 kinase activity levels increased after 8-12 h in cells exposed to CGF and in positive controls exposed to 10% FBS. They returned to basal level 4 h later in cells exposed to CGF alone, whereas in the presence of CGF + EGF and FBS they remained elevated for up to 20 h. The cyclin E levels did not increase in the presence of EGF alone. Cyclin dependent kinase inhibitors p21cip1 and p27kip1 were barely detectable in these cells. Fibroblasts transfected with LXSN-cyclin E, a retroviral vector containing cyclin E cDNA, overexpressed cyclin E and their steady-state cyclin E-Cdk activity was higher than control cells. DNA synthesis by cyclin E overexpressing cells was higher, but optimal DNA synthesis by these cells required the presence of CGF and EGF. These results show that CGF action involves an increase in the levels of cyclin E and E-Cdk activity and that the higher levels are maintained in the presence of both CGF and EGF. They also indicate that sustained high cyclin E levels and Cdk2 activity during G1 phase are necessary, but not sufficient, for optimal mitogenic response in human fibroblasts. PMID- 9737727 TI - CD24 mediates rolling of breast carcinoma cells on P-selectin. AB - P-selectin mediates rolling of neutrophils and other leukocytes on activated endothelial cells and platelets through binding to P-selectin glycoprotein ligand 1 (PSGL-1). Certain PSGL-1 negative tumor cell lines can bind P-selectin under static conditions through the GPI-linked surface mucin, CD24, but the physiological significance of this interaction and whether it can occur under flow conditions is not known. Here, we show that CD24+ PSGL-1- KS breast carcinoma cells attach to and roll on recombinant P-selectin under a continuous wall shear stress, although at a lower density and higher velocity than CD24+ PSGL-1+ cells, such as HL-60. Adding excess soluble CD24 or removing CD24 from the cell surface with phosphatidylinositol-phospholipase C (PI-PLC) significantly reduced KS cell rolling on P-selectin. The ability of KS cells to roll on P selectin was positively correlated with the CD24 expression level. Comparison with three other CD24+ cell lines established that expression of sialyl-Lewis(x) antigen was also necessary for CD24-mediated rolling on P-selectin. CD24 purified from KS cells supported rolling of P-selectin transfectants, but not L-selectin transfectants. Finally, KS cells rolled on vascular endothelium in vivo in a P selectin-dependent manner. Together our data show that CD24 serves as a ligand for P-selectin under physiological flow conditions. Interaction of tumor cells with P-selectin via CD24 may be an important adhesion pathway in cancer metastasis. PMID- 9737729 TI - Integrative round table: introduction. PMID- 9737728 TI - A commentary on the evolution of NIH: what's in, what's out, what's hot, what's not. PMID- 9737730 TI - The challenge and satisfaction of integrative thinking. PMID- 9737731 TI - Ways of integration. PMID- 9737732 TI - The neurobiology of multimodal sensory integration. PMID- 9737733 TI - Decoding the language of the heart: developing a physiology of inclusion. AB - Constructs such as homeostasis and fight/flight have supported a scientific approach to physiology that has yielded a vast database of obvious heuristic value. Yet in spite of its value, these constructs have tended to create a mind set that unwittingly supports what this article has labeled a "physiology of exclusion." Reinforced by the philosophy of Rene Descartes, this perspective has led investigators to focus on isolated or separate animal organisms that are reflexively wired for self-preservation. It has created a mind-set in which both research investigators and the public at large tend to view the human body as either in a steady state of vigilance, maximally prepared for fight/flight, or in a state of quiescence. Assumptions of the solitary body, and solitary man wired to react for "self" preservation, has made it difficult to incorporate a growing body of evidence that indicates that social support and loving relationships are conducive to good health. It also has made it difficult for investigators to fully understand why human loneliness is a major cause of premature death. This article delineates these trends and offers a new construct, one that suggests that a "physiology of inclusion" be added to the prevailing view of a "physiology of exclusion." Recent cardiovascular research is cited to help underscore the potential heuristic value of this new physiological construct. PMID- 9737734 TI - Psychoneuroimmunology: the final hurdle. PMID- 9737735 TI - Integrative thinking: the essence of good medical education and practice. AB - Integrative medicine implies the ability to extract bits of information from seemingly disparate disciplines, and synthesize them into something that is meaningful. Contemporary medical research is more apt to be driven by purely commercial incentives that not only do not favor integrative efforts, but deliberately suppress them when vested interests are threatened. Good health depends entirely on good communication whenever homeostasis is threatened. This requires a continual and instantaneous exchange of information between the constituency of the internal environment that can only be comprehended by an integrative approach. Various examples are given to illustrate these and other relevant issues. PMID- 9737737 TI - To the centenary of P.K. Anokhin, a great Russian physiologist. PMID- 9737736 TI - The impact of a new emotional self-management program on stress, emotions, heart rate variability, DHEA and cortisol. AB - This study examined the effects on healthy adults of a new emotional self management program, consisting of two key techniques, "Cut-Thru" and the "Heart Lock-In." These techniques are designed to eliminate negative thought loops and promote sustained positive emotional states. The hypotheses were that training and practice in these techniques would yield lowered levels of stress and negative emotion and cortisol, while resulting in increased positive emotion and DHEA levels over a one-month period. In addition, we hypothesized that increased coherence in heart rate variability patterns would be observed during the practice of the techniques. Forty-five healthy adults participated in the study, fifteen of whom acted as a comparison group for the psychological measures. Salivary DHEA/DHEAS and cortisol levels were measured, autonomic nervous system function was assessed by heart rate variability analysis, and emotions were measured using a psychological questionnaire. Individuals in the experimental group were assessed before and four weeks after receiving training in the self management techniques. The experimental group experienced significant increases in the positive affect scales of Caring and Vigor and significant decreases in the negative affect scales of Guilt, Hostility, Burnout, Anxiety and Stress Effects, while no significant changes were seen in the comparison group. There was a mean 23 percent reduction in cortisol and a 100 percent increase in DHEA/DHEAS in the experimental group. DHEA was significantly and positively related to the affective state Warmheartedness, whereas cortisol was significantly and positively related to Stress Effects. Increased coherence in heart rate variability patterns was measured in 80 percent of the experimental group during the use of the techniques. The results suggest that techniques designed to eliminate negative thought loops can have important positive effects on stress, emotions and key physiological systems. The implications are that relatively inexpensive interventions may dramatically and positively impact individuals' health and well-being. Thus, individuals may have greater control over their minds, bodies and health than previously suspected. PMID- 9737738 TI - Electroencephalographic biofeedback methodology and the management of epilepsy. AB - Currently considerable research is being directed toward developing methodologies for controlling internal processes. An applied branch of the basic field of psychophysiology, known as biofeedback, has developed to fulfill clinical needs related to such control. Current scientific and popular literature abounds with numerous examples of how biofeedback is being used. For example, germinal studies by Kamiya (1962), and later work by Lynch and Paskewitz (1971), Beatty (1973), as well as many others have shown that the EEG alpha rhythm (8 to 13 Hz) recorded from occipital regions of the human brain can be behaviorally manipulated when feedback or reward is provided for changing the density of this activity. Other researchers have provided evidence that theta activity (4 to 7 Hz) and the beta activity (greater than 14 Hz) can also be controlled by humans and analogs of this activity have been conditioned in animals as well (Green, Green and Walters, 1971). In addition to the work that has been carried out with the EEG, researchers such as Engle and Bleecker (1973) have indicated that it might be possible to control cardiac arrhythmias through biofeedback. Studies by Elder et al. (1973) have provided some hope that blood pressure in humans might also be conditioned. Also, considerable effort has been directed to the control of responses from single muscles with particular applied emphasis in neuromuscular rehabilitation, control of muscle tension for tension headaches and the management of migraine headaches through vasomotor conditioning (Brudny et al., 1974; Basmajian, 1963, 1971; Sargent et al., 1973). PMID- 9737739 TI - Demonstration of extracellular acid phosphatase activity in the involuting, antimesometrial decidua in fed and acutely fasted mice by combined cytochemistry and electron microscopy. AB - An ultrastructural cytochemical study of acid phosphatase activity in the antimesometrial decidua on days 9-11 of pregnancy was performed in fed and acutely fasted mice. Specimens were fixed in a buffered mixture of paraformaldehyde and glutaraldehyde and were incubated in a buffered medium containing sodium beta-glycerophosphate and cerium chloride for ultrastructural localization of acid phosphatase activity. Fed and fasted animals showed extracellular acid phosphatase reaction product in the decidual-trophoblast interface, in the region of loosely and tightly packed, mature decidual cells, and in the region of predecidual cells. Reaction product was absent in the region of nondecidualized stromal cells. Extracellular acid phosphatase activity was more conspicuous in the region of mature decidual cells in fasted mice than in fed mice, and it was apparently similar in the region of predecidual cells in both fed and fasted mice. Acid phosphatase reaction product was also observed in lysosomes in all cells studied. Because acid phosphatase activity reflects the presence of lysosomal hydrolases in general, our results suggest that there is matrix degradation by lysosomal enzymes in both fed and fasted mice. These events may be part of the process of tissue remodeling in regions of predecidual cells and mature decidual cells. However, it is also possible that, in the region of mature decidual cells, breakdown of matrix constituents is a mechanism to provide nutrients for the growing fetus. This mechanism is probably enhanced in fasted mice. PMID- 9737740 TI - Stages of the cycle of the seminiferous epithelium of the viscacha (Lagostomus maximus maximus). AB - The adult male viscacha (Lagostomus maximus maximus) is a seasonal rodent. It exhibits a short period of testicular regression with partial arrest of spermatogenesis during winter (July-August). The present study provides the first description of the viscacha spermatogenic cycle during the period of maximum gonadal activity (summer-autumn). The testes were processed by using conventional techniques of light and electron microscopy. One-micrometer-thick sections stained with toluidine blue were used to clearly define the different cell associations. Spermatogenesis in this rodent is well organized and synchronized and has been divided into nine stages. The present classification is based on the morphogenesis of the acrosomal system (stages I-V) and the degree of elongation and condensation of the heads of the differentiating spermatids (stages VI-IX). Stage I is principally defined by round spermatids with a well-developed, juxtanuclear Golgi complex and without acrosomal components that are visible under the light microscope. Sperm release from the viscacha seminiferous epithelium occurs either in late stage III or in early stage IV. Stage IX is characterized by diplotene spermatocytes, figures of meiosis I or II, and secondary spermatocytes. PMID- 9737741 TI - Neonatal exposure of male rats to estradiol benzoate causes rete testis dilation and backflow impairment of spermatogenesis. AB - Estrogens administered to perinatal rodents cause spermatogenesis impairment; this study was undertaken to determine the mechanisms by which estrogens exert this effect. Neonatal male Wistar rats received estradiol benzoate (either 0.5 mg/5g BW or 1 mg/5g BW) and were killed at days 10, 22, 33, 45, and 60. Controls received vehicle. In tubule cross-sections of transverse sections of the right testes, 1) tubular diameter (TD) and seminiferous epithelium height (SEH) were measured, 2) normal and impaired spermatogenesis were classified in terms of the most advanced germ cell type present, including tubules lined by Sertoli cells only. A significant dose-dependent rise in the tubule percentage lined by Sertoli cells only at day 60 reflected spermatogenesis impairment. This was evidenced by the presence of multinucleated germ cells in a thin epithelium and sloughed into an enlarged tubular lumen, which was reflected in a significant dose-dependent increase in TD/SEH values from day 22 onward. TD was significantly greater and SEH significantly lower in tubular segments located at the cranial than the caudal halves of rat testes treated with the high (days 22, 33, and 60) and the low dose (day 33). This indicated distension in cranial tubular segments, perhaps due to the fact that these segments were the closest to the dilated rete testis. Consequently, they showed the highest TD/SEH values and the most regressive features of spermatogenesis (tubules lined by Sertoli cells only). In contrast, caudal segments in rat testes treated with the low dose showing TD/SEH values similar to controls displayed a delayed maturation of spermatogenesis coinciding with the late appearance of mature Leydig cells. PMID- 9737742 TI - Proportions of slow-twitch and fast-twitch extrafusal fibers in receptive fields of tendon organs in chicken leg muscles. AB - Golgi tendon organs are mechanoreceptors that monitor the contractile force produced by motor units. Receptors are most responsive to contractions of extrafusal muscle fibers that terminate closest to them and on them. Three anterior and four posterior chicken leg muscles were examined. Proportions of immunohistochemically identified slow-twitch extrafusal fibers and fast-twitch extrafusal fibers were calculated for 374 tendon organ receptive fields. Tendon organs were observed in muscle regions occupied either by slow-twitch fibers or fast-twitch fibers only, but most were found in regions that contained both slow twitch and fast-twitch extrafusal fibers. The frequency with which each fiber type occurred near tendon organs approached the frequency with which it occurred in more inclusive regions. In receptive fields with mixed fiber populations, fast twitch fibers were the predominant type, especially in the anterior leg muscles. Distribution patterns of extrafusal fiber types adjacent to and farther removed from tendon organs suggest that afferent discharges from tendon organs are by and large unbiased measures of the contractile activity of the extrafusal fiber population of the muscle portion in which the tendon organs are located. In mixed muscle regions, slow-twitch fibers and fast-twitch fibers attach on given tendon organs, enabling them to monitor forces produced by slow motor units and by fast motor units. Most tendon organs are situated in mixed extrafusal fiber fields with high fast-twitch fiber content, indicating that in chicken leg muscles sensory feedback from tendon organs is largely one from fast motor units. PMID- 9737743 TI - Proliferation of goblet cells and vacuolated cells in the rabbit distal colon. AB - Previous studies of colonic epithelial cell kinetics in mice and rats revealed a pattern similar to small intestine, where basally located stem cells proliferate, differentiating as they migrate towards the surface epithelium. Vacuolated and goblet cells are assumed to co-migrate at the same rate. The present study indicates that rabbit distal colon has more complicated epithelial cell kinetics. The zone of proliferation was detected immunohistochemically using proliferating cell nuclear antigen (PCNA) and confirmed with the use of colchicine to arrest dividing cells in metaphase. Migrating cells were tracked from the zero-hour position (PCNA labeling, mitosis) to positions 24, 48, 72 hrs by monitoring cell migration with the thymidine analog 5-Bromo-2-Deoxyuridine (BrdU). PCNA revealed a major proliferative zone in the upper third of the crypt column and the presence of mitotic figures after colchicine corroborated these results. Differentiated vacuolated cell proliferation was detected at three crypt sites: base, middle, and top of the crypt, while columnar cells arose from a population of dividing cells at the top of the crypt. Turnover of columnar and vacuolated cells occurred within 72 hrs. Goblet cells exhibited maximal proliferation at the crypt base and migrated at a much slower rate than the other cell types. In rabbit distal colon, populations of proliferating cells exist at multiple levels of the crypt column. Vacuolated and goblet cells differ in their labeling indices and migration rates, suggesting that the two cell types arise and migrate independently. PMID- 9737744 TI - Capillary growth in overloaded, hypertrophic adult rat skeletal muscle: an ultrastructural study. AB - We examined the early stages of angiogenesis in overloaded m. extensor digitorum longus following extirpation of the agonist m. tibialis anterior. Capillary-to fibre ratio increased after 1 week (1.54+/-0.02) vs. control (1.38+/0.06; P < 0.01) and resulted in a greater tortuosity of the capillary bed at 2 weeks, indicating the presence of lateral sprouts or anastomoses. Capillary endothelial cells (ECs) showed ultrastructural signs of activation, were thickened, and had irregular luminal and abluminal surfaces. The proportion of ECs with abluminal processes increased after overload (13.5+/-0.6% vs. 2.0+/-1.5%, 1 week vs. contralateral, P < 0.01; 12.5+/-2.6% vs. 3.5+/-0.6%, 2 weeks vs. contralateral, P < 0.01), whereas there was no significant change in proportion of luminal processes. Abluminal processes occurred in approximately 13% of capillaries in overloaded muscles (P < 0.01 v. control and contralateral), and most were associated with focal breakage of the basement membrane (BM). Small sprouts (<3 microm in diameter) comprised of one or two ECs sometimes lacked a lumen, and others had a slitlike or vacuolelike lumen between adjacent ECs or vacuolelike lumen formed by fusion of vesicles within a single EC. Endothelial mitosis was occasionally seen in nonsprouting capillaries with intact BM, increasing the average number of ECs per capillary from approximately 1.7 in control muscles to 2.1 after 1 week of overload (P < 0.05) when bromodeoxyuridine incorporation was also higher (P < 0.001). We conclude that muscle overload induces capillary growth by sprouting of existing capillaries, probably due to mechanical stretch acting from the abluminal side of the vessels. PMID- 9737745 TI - Comparative ultrastructure of intercalated ducts in major salivary glands: a review. PMID- 9737746 TI - Computerized morphometry of the pulmonary vasculature over a range of intravascular pressures. AB - Recent availability of computerized image analysis has fostered hope that barium injection and landmarking of pulmonary arteries would be unnecessary for morphometric assessment when using this technique. We reasoned that if barium injection altered morphometric variables, it would do so in a linear fashion correlating with incremental increases in injection pressure of the barium. The two goals of the present study were to determine whether barium injection into arteries affected morphometric measurements and to determine whether incremental increases in injection pressure correlated with alterations in morphometric measurements in a linear fashion. Computerized image analysis was used to measure the internal elastic lamina (IEL) and external elastic lamina (EEL). Medial area (MA), luminal area (LA), percentage of medial thickness, IEL square root of MA, and idealized LA were calculated. Barium injection did not alter morphometric variables in a linear fashion correlating with incremental increases in injection pressure of the barium except the percentage of arteries that filled with barium. Maximum recruitment for pre-acinar arteries occurred at 40 mmHg pressure and 60 mmHg distending pressure for intra-acinar arteries. Incremental increases in injection pressure did not affect IEL, EEL, or calculated morphometric variables. However, IEL, medial thickness, and MA were all smaller in injected vessels than in uninjected vessels. IEL square root of MA and the ratio of measured vs. idealized LA were both increased in injected lungs. We suspect that vascular injection selects for evaluation, a population of smaller, thin-walled vessels, which in the uninjected lungs are collapsed and hence excluded from analysis. PMID- 9737747 TI - Tongue flicking in agamid lizards: morphology, kinematics, and muscle activity patterns. AB - We wanted to examine whether a relation between foraging strategy, morphology, the mechanics of tongue protrusion, and prey chemical detection and discrimination exists in agamid lizards. Tongue-flick behavior was observed in two species of this family: Uromastix acanthinurus and Plocederma stellio. Potential prey chemical discrimination by means of tongue flicking was examined by using applicator tests. Tongue flicks were subsequently recorded by high-speed video in combination with the electrical activity of a number of jaw and hyolingual muscles. The kinematics of jaws and tongue and the muscle activity patterns were quantified. To investigate if the observed differences in tongue flick behavior (mainly in the frequency of use) are translated into corresponding differences in tongue morphology, the tongues of both species were examined by light and scanning electron microscopy. The species differed mainly in the surface morphology of the foretongue and in the abundance and distribution of taste buds on the tongue and oral cavity. These differences can be related to behavioural observations; whereas U. acanthinurus readily uses tongue flicks to detect and discriminate between food items, P. stellio does not. However, differences in tongue-flick mechanics (kinematics, electromyograms) between both species were minor. Based on the data gathered in this study and from previously published data, an evolutionary transformation series leading to the complex tongue-flick cycles as observed in snakes is proposed. The required morphological and mechanical changes that accompany such an evolutionary sequence are discussed. PMID- 9737748 TI - Immunolocalization of the cleavage of the aggrecan core protein at the Asn341 Phe342 bond, as an indicator of the location of the metalloproteinases active in the lysis of the rat growth plate. AB - In view of the extensive lysis of hyaline cartilage known to take place during endochondral bone formation, the current study was designed to test the hypothesis that metalloproteinases are the agents that mediate this lysis. Since these enzymes have been shown in vitro to cleave the core protein of the major proteoglycan of cartilage, aggrecan, at the Asn341-Phe342 bond, an immunohistochemical method has been developed to find out whether or not there are sites in the growth plate of the rat tibia where cleavage of this bond takes place. The cleavage of aggrecan by metalloproteinases is followed by the retention of the fragment known as G1, for it includes the G1 domain. Since the G1 fragment terminates in the amino acid residues ...FVDIPEN, we prepared an antiserum against FVDIPEN, confirmed its specificity, then applied it to the growth plate of 21-day-old rat tibia in the hope of localizing the G1 fragments. The antiserum specificity was shown by its recognition of the ...FVDIPEN sequence at the C-terminus of peptides and of G1 fragments produced by aggrecan cleavage. When the antiserum was applied to Western blots of guanidinium chloride extracts prepared from epiphyseal growth plate, it recognized two species (56 and 52 kDa), which differed only in the degree of glycosylation. These fragments were comparable in size to the G1 fragments generated by the action of recombinant metalloproteinase in vitro, thus confirming antiserum specificity for these fragments. Applying the antiserum to cryosections of 21-day-old rat tibiae revealed immunostaining at two intensities within the growth plate matrix: a strong staining was observed in a 1-5 microm-wide layer designated "peripheral" matrix, which borders the epiphyseal and metaphyseal marrow spaces as well as the perichondrium, while a weak staining was found in the rest of the plate, designated "central" matrix. The abundance of G1 fragments terminating in ...FVDIPEN in the peripheral matrix indicates that this is where the growth plate is lysed to achieve longitudinal and latitudinal bone growth. The site where metalloproteinases exert their main lytic activity is a thin layer of matrix separating central from peripheral matrix. PMID- 9737749 TI - Photoreceptor morphogenesis in the human retina: a scanning electron microscopic study. AB - There are a number of scanning electron microscopic (SEM) studies on retinal photoreceptors of vertebrates. However, most of these are concerned with the adult retina, and only a very few deal with developing photoreceptors. In man, SEM studies have not been carried out on photoreceptor morphogenesis during fetal or postnatal stages. Hence, the present study was undertaken to examine the sequential morphological changes in developing photoreceptors during different gestational ages in the human retina. Retinas of human fetuses of gestational ages of 10-25 weeks and from autopsy of a 5-month-old infant were processed for SEM. The observations show some new information on the morphogenesis of photoreceptors. At 10-11 weeks, the outer and inner neuroblastic zones are well developed and separated from each other by the layer of Chievitz. By 15-16 weeks, the photoreceptor precursors appear as spherical inner segments on the scleral surface of the outer neuroblastic zone. Cilia develop as small protrusions from the apical ends of the inner segments. Photoreceptor inner segments become arranged in mosaic pattern by 18-19 weeks. In the mosaic, large cone inner segments (putative blue cones) stand out prominently from the remaining small cone inner segments (prospective red/green cones). The rod inner segments are identifiable and show cilia. Between 19-20 and 24-25 weeks, the cone inner segments elongate and change in shape from spherical to oval. At 24-25 weeks, the outer segments develop from the distal ends of rod cilia. At this period, the inner segments of rods and cones are interconnected by protoplasmic projections. Although the precursors of both rods and cones appear to be in a similar state of development at 14-15 weeks gestation, the rods undergo morphological maturation earlier than do the cones. Photoreceptor development in the anterior retina lags behind that of the posterior retina by about 10 weeks. At 5 months after birth, the posterior retina possesses fully developed photoreceptors that are comparable to those of the adult. However, the photoreceptors in the ora serrata resemble those in the posterior retina of 24-25 weeks gestation. PMID- 9737750 TI - Epiphyseal and physeal cartilage vascularization: a light microscopic and tritiated thymidine autoradiographic study of cartilage canals in newborn and young postnatal rabbit bone. AB - The vascular pattern of newborn and early postnatal epiphyseal and physeal cartilage is integral to long bone development and differs from later postnatal patterns. In the present study, we supplement light microscopic histology with tritiated thymidine autoradiography to help assess the position of cartilage canals and the dynamics of cartilage vascularity in relation to growth. Tritiated thymidine labeling studies to assess cell proliferation activity were done by using 2 microc/g body weight intraperitoneal injections into newborn and 3-, 4-, and 7-day-old New Zealand white rabbits that were killed 1 hr after the injection. Proximal humeral, distal femoral, and third metatarsal epiphyses were assessed by routine histology and serial section autoradiography. Cartilage canals were seen in each epiphysis. Transphyseal vessels were seen in each epiphysis continuous from the epiphysis to the metaphysis or were present within the physis traversing the proliferating and hypertrophic cell zones. Histologic sections showed vessels from the perichondrium continuous with those of the epiphyseal cartilage canals at proximal humeral, distal femoral, and metatarsal epiphyses. Serial sections showed vascular buds and connective tissue cells lying in indentations at the periphery of and present within the epiphyseal cartilage. Autoradiographic studies showed extensive labeling of vessel wall cells and surrounding connective tissue cells of the cartilage canals (a) within the epiphyseal cartilage, (b) traversing the physis, and (c) within the epiphyseal cartilage but continuous with the perichondrial vessels. The labeling was always far more extensive than in the surrounding chondrocytes and was always present throughout the entire extent of the canals. In conclusion, the cell labeling activity strongly supports an active dynamic phenomenon underlying the vascularization of epiphyseal and physeal cartilage. PMID- 9737751 TI - Lectin staining in the basal nucleus (Meynert) and the hypothalamic tuberomamillary nucleus of the developing human prosencephalon. AB - Previous studies have demonstrated that extracellular matrix glycoconjugates, shown by lectin-histochemistry with Vicia villosa agglutinin (VVA) and peanut agglutinin (PNA) as so-called perineuronal nets, play an important role in brain maturation. Concanavalin A (ConA) binding to neuronal surface glycoconjugates may be a marker of synaptic junctions. The present study was done to demonstrate the binding sites of these lectins in two functionally related nuclei of the prosencephalon, the basal nucleus (Meynert) and the hypothalamic tuberomamillary nucleus. Fetal brains of 16-36 weeks of gestation were examined by using VVA, PNA, and ConA to determine appearance and distribution patterns of specific lectin-binding sites on glycoconjugates during fetal brain development. The basal nucleus and the tuberomamillary nucleus showed a characteristic "cellular staining" that may have been due to cytoplasmatic labeling, surface labeling, or both. Lectin-staining occurred much earlier in the basal nucleus than in the tuberomamillary nucleus. Although all three lectins were bound to neurons of the basal nucleus, only ConA-positive neurons were observed in the tuberomamillary nucleus. In conclusion, lectin-labeled cells most probably represent projection neurons that are GABAergic (tuberomamillary nucleus) or cholinergic (basal nucleus). Labeling with the three lectins demonstrated nuclear-specific staining patterns that occur early in fetal development and gradually increase. Binding sites for lectins characterizing perineuronal nets (VVA, PNA) occurred only in the basal nucleus, whereas binding sites for ConA on neuronal-surface glycoconjugates, which seem to play a role in early synaptogenesis, were present in the basal and the tuberomamillary nucleus. The basal nucleus, however, expressed ConA binding sites distinctly earlier, probably indicating early arriving afferents. PMID- 9737752 TI - Right axis deviation as a criterion for echocardiographic evaluation of aircrew candidates. AB - BACKGROUND: Significant pressures exist in the resource-limited environment of military aviation to select only those candidates for flight training who are both physically able to withstand the demands of the flight environment and likely to complete a career in aviation. As medical technology has advanced, uncertainty has arisen regarding the appropriate use of additional studies in the screening of aircrew candidates. At the Naval Operational Medicine Institute (NOMI), where all U.S. Navy aviation duty candidates' medical qualifications are reviewed, current policy is to perform 2D-echocardiography on all candidates who exhibit a right axis deviation of +95 degrees or greater on their initial electrocardiogram. METHODS: The records of all aircrew candidates referred to the Department of Internal Medicine for evaluation of right axis deviation during the years 1993, 1994, and 1995 were reviewed. A cost benefit analysis was performed to assess the cost effectiveness of using echocardiography to screen candidates suspected of having disqualifying physical defects based on a finding of right axis deviation on electrocardiogram. RESULTS: Of the 69 cases reviewed, only 1 candidate was disqualified due to cardiovascular disease. He suffered from a large atrial septal defect initially discovered on physical examination. CONCLUSIONS: It is not cost effective to use 2D-echocardiography to screen aircrew candidates for disqualifying cardiac defects based on a finding of a right axis deviation of +95 degrees or greater on electrocardiogram. PMID- 9737754 TI - Comparison of responses of men to immersion in circulating water at 40.0 and 41.5 degrees C. AB - BACKGROUND: The recommended maximum water temperature for public hot tubs has been set at 40.0 degrees C, but no research has been published on human immersion in hot water at higher temperatures. HYPOTHESIS: We hypothesized that thermoregulatory and cardiovascular responses at two water temperatures would be proportional to the water:blood temperature gradients. METHODS: Six healthy men were immersed for 21 min in circulating hot water at 40.0 and 41.5 degrees C in separate trials in random order 1-3 wk apart. Measurements included heart rate, systolic BP, esophageal, rectal, and non-immersed skin temperatures, sweat rate, and perceived comfort. RESULTS: The rise in all body temperatures, sweat rate, and heart rate were significantly greater in the 41.5 vs. 40.0 degrees C water. Peak esophageal temperatures were 38.3 +/- 0.2 degrees C vs. 37.8 +/- 0.03 degrees C, peak sweat rates were 0.48 +/- 0.05 vs. 0.32 +/- 0.03 kg x m(-2) x h( 1), and peak heart rates were 123 +/- 7 vs. 108 +/- 5 bpm, respectively. Systolic BPs followed different patterns of response in each trial, whereas diastolic pressures were not different between trials. Comfort at each level of immersion was reduced during the 41.5 degrees C trial compared with the 40.0 degrees C in excess of that predicted by difference in esophageal temperature between the trials. CONCLUSIONS: These results suggest that risks of hyperthermia or adverse cardiovascular effects in hot tubs may not be greater in water above 40.0 degrees C unless perceptual judgment is impaired. Hypotension when standing to exit the tub occurred in both trials and may represent a potential hazard to hot tub use. PMID- 9737753 TI - Plasma volume expansion with oral fluids in hypohydrated men at rest and during exercise. AB - BACKGROUND: The purpose for this study was to evaluate various carbohydrate (CHO) electrolyte fluid formulations for consumption by astronauts to maintain or restore their plasma volume (PV) and total body water (TBW) during and after extravehicular activity (exercise experiment, EE) and for a few hours before reentry and immediately after landing (rest experiment RE). HYPOTHESIS: That fluid formulation electrolyte content would be more important than osmotic (Osm) content for increasing or maintaining PV during the RE and EE. METHODS: In the RE, 5 healthy men (23-44 yr), previously dehydrated for 24 h, drank 6 fluid formulations (Water, 19.6 Na, 157 Na, 19.6 Na + glucose, and the prepared drinks Performances and Power)--one each at weekly intervals, and then sat for 70 min. In the EE, four healthy 24-h dehydrated men (30-46 yr) exercised for 70 min supine on a cycle ergometer (load = 71 +/- 1% peak VO2). RESULTS: Rest: Subjects who consumed formulations with total Osm concentrations nearer the normal range (157 Na - 270 mOsm x kg(-1), Performance with 19.6 mEq x L(-1) Na - 380 mOsm, and to some extent Power with 23.5 mEq x L(-1) Na - 390 mOsm) had the greater increases in PV; intake of drink 157 Na, with the largest Na content, induced the greatest hypervolemia of 7.6% (p < 0.05). The various additional ions, in addition to 19.6 Na, probably contributed to the 4.6% (p < 0.05) hypervolemia with Performance. Water was not effective. Exercise: Stabilization of PV between 15-70 min was not related to drink total CHO, Na or Osm content. Performance and 157 Na were no more effective than 19.6 Na or 19.6 Na + glu for PV stabilization. Water was the least effective. Regulatory mechanisms controlling PV during exercise appear to be independent of oral fluid formulation Osm-electrolyte content. CONCLUSIONS: Drink cation (sodium) content is more important that its total osmotic content for increasing plasma volume at rest. Fluid formulations with greater hypervolemic action in resting subjects may not be as effective during exercise; therefore different formulations for use during exercise appear to be necessary. PMID- 9737755 TI - Rhinovirus replication in HeLa cells cultured under conditions of simulated microgravity. AB - BACKGROUND: Rotating-wall vessels (RWVs) allow for the growth of cells under conditions of simulated microgravity. Information about the replication of viruses in simulated microgravity using RWVs has not been reported. Cells grown in RWVs are subjected to low shear motion, and the replication of certain viruses such as rhinoviruses has been reported to be enhanced by motion. HYPOTHESIS: Our research was based on the hypothesis that rhinovirus replication would be enhanced under conditions of simulated microgravity. METHODS: HeLa cells were cultured in three-dimensional cultures on microcarrier beads in simulated microgravity using RWVs and in sealed Teflon roller bottles. Two-dimensional cultures of HeLa cells were also grown in tissue culture flasks (T-150s). Viral infections for all cultures were carried out under standardized conditions at 1 x g. The amount of new virus released during the first viral replication cycle and the total viral yields obtained from multiple viral replication cycles were determined. RESULTS: Viral quantitation during the first viral replication cycle showed that after 10-13 h RWV and Teflon roller bottle supernatants contained significantly more virus than the supernatants from T-150 cultures. After multiple viral replication cycles (at 24, 48, 72, and 96 h following infection), total viral samples (both free and cell-associated virus) from RWV cultures contained significantly more virus than Teflon roller bottle cultures. CONCLUSIONS: The rhinovirus replication cycle was enhanced in cultures grown in the presence of motion (Teflon roller bottle cultures and RWV cultures). Additionally, multiple rounds of rhinovirus replication yielded more virus in simulated microgravity conditions. Viral transmission in cell cultures in RWVs was efficient and was similar to or better than what occurred in the Teflon roller bottles. The cultivation of cells in simulated microgravity possibly affected the rate of viral adsorption/uptake, the viral replication cycle, and/or the viral yield. RWVs provide an effective means for culturing human rhinoviruses. PMID- 9737756 TI - Female exposure to high G: echocardiographic evaluation for chronic changes in cardiac function. AB - BACKGROUND: Significant changes in cardiac preload and afterload are generated by a number of factors present during the operation of high performance aircraft. These include high levels of +Gz, positive pressure breathing and anti-G straining maneuvers. Centrifuge subjects are exposed to these same factors in doses that are comparable to their operational counterparts. The question of whether such exposures produce long-term adverse effects on the heart has not been definitively answered. METHODS: In an effort to further address this issue, a longitudinal study was conducted on 18 newly recruited centrifuge panel members (7 males and 11 females) who did not have a previous history of significant high +Gz exposure. In order to document the cumulative long-term effects of high +Gz exposure and G protection measures, baseline echocardiographic studies were conducted prior to any +Gz exposure on the Dynamic Environment Simulator (DES) centrifuge. The echocardiograms were repeated after each panel member completed eight sessions of indoctrination. These follow-up echos were performed after all 18 subjects had been exposed to over 45 min (cumulative) of sustained acceleration > or = 2 G. Each subject served as his/her own control. All studies were evaluated independently by a cardiologist who was blinded to the order in which the studies were performed. Although complete echocardiographic studies were performed, only the parameters identified as significant in prior studies were evaluated. RESULTS: No significant differences were found between the initial and follow-up echo parameters. No significant differences were found between male and female responses. CONCLUSIONS: We found no significant differences in cardiac function after 45 min (cumulative) of exposure to G > or = 2 in men or women. These subjects will be monitored during a longitudinal study throughout their centrifuge subject career. PMID- 9737757 TI - Female exposure to high G: performance of simulated flight after 24 hours of sleep deprivation. AB - BACKGROUND: Ground-based research has investigated the loss of cognitive function in the extreme conditions of G-induced loss of consciousness, however, little is known about pilots' abilities to maintain cognitive performance throughout prolonged conscious exposure in the high-G environment. The effects of fatigue and G layoff on performance during exposure to high G are mostly unknown for the female population. METHODS: This research was conducted on the centrifuge Dynamic Environment Simulator. Active-duty personnel (8 male and 8 female) were trained to fly the F-16 simulation while 30 performance measures were recorded. Performance was re-evaluated after 24 h of sleep deprivation. RESULTS: Neither male nor female overall performance was affected significantly by sleep status, although individual tasks showed sensitivity; call-sign reaction time was longer by 33%, and missile survival was less likely. Also, when sleep deprived, perceived effort and physical demand were higher while perceived performance was lower. No differences in performance were found in either gender due to lay-off, although some physiologic deconditioning was apparent. Women commanded and endured the same amount of G load as men, however, on average they could not perform the tracking task quite as well. CONCLUSIONS: Sleep deprivation (24 h) produced sensations of fatigue and frustration, but overall performance was not reduced. The ability of personnel to complete a complex defensive maneuver was reduced when they were sleep deprived. The women that we tested apparently could not optimize the tracking task as well as their male counterparts when Gz was in the simulation. None of these results were sufficient to suggest that women should not be allowed to compete for flying assignments in high-performance aircraft. PMID- 9737758 TI - Female exposure to high G: effects of simulated combat sorties on cerebral and arterial O2 saturation. AB - BACKGROUND: One of the key factors in maintaining optimal cognitive performance in the high-G environment is the adequate delivery of oxygen to the cerebral tissue. As eye-level blood pressure is compromised at 22 mmHg x G(-1), perfusion to the peripheral cerebral tissues (cerebral cortex) may not be adequate to support the mental demands of flight. This study measured the effect of closed loop flight simulations (3 min) on cerebral oxygen saturation changes (rSO2), arterial oxygen saturation (SAO2), and heart rate (HR), in both rested (8 h of rest) and sleepless (24 h without sleep) conditions. METHODS: Subjects (16; 8 males and 8 females) were subjected to G-exposures via closed-loop flight simulations in a series of four 3-min sorties flown by subjects on the Dynamic Environment Simulator (centrifuge) in either a rested or a sleepless state. Prior to the centrifuge flight, subjects were instrumented with sensors for measurement of arterial oxygen saturation (SAO2) and regional cerebral tissue oxygenation (rSO2). Subjects wore the standard flight suit, boots, CSU-13B/P anti-G suit, and the COMBAT EDGE positive-pressure breathing for G-protection system. RESULTS: Significant changes in cerebral and arterial oxygen saturation were observed within groups when comparing pretest baselines and minimum values during the test and pre- and post-G rSO2, SAO2, and HR in both the rested and sleepless state, (p # 0.01), respectively, for each group. Comparisons between groups showed women to have significantly smaller regional cerebral cortex oxygen decreases than men (p # 0.01). No significant changes in SAO2, however, were observed between groups. Both men and women showed a slow recovery of rSO2 values to the prebaseline levels. CONCLUSIONS: Sleeplessness had no effect on the rSO2, SAO2, and HR compared with the rested condition. During acceleration, regional cerebral tissue oxygen decreased 13% in men compared with 9% in women. The recovery of cerebral tissue oxygen levels to prebaseline values was retarded somewhat when compared with the recovery response of arterial oxygen saturation. PMID- 9737759 TI - Female exposure to high G: chronic adaptations of cardiovascular functions. AB - INTRODUCTION: Exposure to microgravity is associated with increased leg venous compliance and reductions in cardiac output, baroreflex functions, and tolerance to orthostatism. However, the effects of chronic exposure to high-G environments are unknown. In addition, there is evidence that females have lower orthostatic tolerance than males, although the underlying mechanisms are unclear. Therefore, we tested the hypotheses that high-G training will enhance baroreflex and orthostatic functions and that females will demonstrate similar adaptations compared with males. METHODS: Calf venous compliance, baroreflex function, and orthostatic performance were measured in six men and seven women before and after repeated exposures on the centrifuge (G-training) for 4 wk, 3 times/wk, with gradual levels of G starting with +3 Gz without G-suit protection during week 1 and advancing to +9 Gz with G-suit protection by the end of week 4. Calf venous compliance was measured by occlusion plethysmography using impedance rheographic recordings of volume change. Baroreflex function was assessed from beat-by-beat changes in heart rate (HR) and mean arterial pressure (MAP) that were measured before, during, and after a Valsalva maneuver strain at 30 mmHg expiratory pressure. The orthostatic performance of reflex responses was assessed from beat by-beat changes in HR, MAP, stroke volume (SV), cardiac output (Q; by impedance plethysmography), and systemic peripheral resistance during the last 10 cardiac beats of a 4-min squat position and during the initial 10 cardiac beats in a standing position. RESULTS: G-training increased calf compliance in both men and women. SV and Q were increased during the squat-to-stand test in the males, but not in the females, following G-training and provided protection against the development of acute hypotension in the men. CONCLUSIONS: G-training caused adaptations in orthostatic functions opposite to those observed following exposure to microgravity environments. However, adaptations to G-training were limited in females, a finding that may provide a physiological basis for their lower simulated combat tracking performance during simulated aerial combat maneuvers compared with males. PMID- 9737760 TI - Acetazolamide plus low-dose dexamethasone is better than acetazolamide alone to ameliorate symptoms of acute mountain sickness. AB - METHODS: In a double-blind study, we compared the efficacy of a combination of sustained-release acetazolamide and low-dose dexamethasone and acetazolamide alone for prophylaxis against acute mountain sickness (AMS) caused by rapid ascent to high altitude. Before ascent, 13 subjects were randomly assigned to receive a combination of one sustained-release acetazolamide capsule (500 mg) in the afternoon and 4 mg dexamethasone every 12 h, or a combination of the same dose of acetazolamide once daily and a placebo every 12 h. Days 1 and 2 were spent at 3698 m (La Paz, Bolivia), while days 3 and 4 were spent at 5334 m (Mount Chaclataya, Bolivia). Ascent was by 2 h motor vehicle ride. Heart rates, peripheral oxygen saturations and a modified score derived from the Environmental Symptom Questionnaire (modified-ESQ) were measured on each day. In addition, weighted averages of the cerebral (AMS-C) and respiratory (AMS-R) symptoms were calculated for days 3 and 4. RESULTS: Heart rate and modified-ESQ scores increased on days 3 and 4 compared with the other days in the acetazolamide/placebo group only (p < 0.05). Oxygen saturations decreased in both groups on days 3 and 4 (p < 0.05), but the decrease was greater in the acetazolamide/placebo group (p < 0.05). AMS-C and AMS-R scores rose above the suggested thresholds for indication of AMS on days 3 and 4 in the acetazolamide/placebo group only (p < 0.05). CONCLUSION: We conclude that this combination of sustained-release acetazolamide once daily and low-dose dexamethasone twice daily is more effective in ameliorating the symptoms of AMS than azetazolamide alone at the ascent that was studied. PMID- 9737761 TI - Evidence that the slowing caused by acute hypoxia is modality dependent. AB - BACKGROUND: AFM (Additive Factors Method) experiments conducted with visual stimuli suggest that the slowing produced by acute hypoxia is located at the earliest preprocessing stage of information processing and that later stages are unaffected (the bottleneck hypothesis). METHODS: To determine the contribution of degraded visual functioning to slowing, we bypassed this modality and measured reaction time in an AFM paradigm to auditory (Experiment 1) and kinesthetic (Experiment 2) stimuli. In both experiments hypoxia was induced with low oxygen mixtures and arterial blood oxygen saturation (SaO2) was controlled at 65%. Task difficulty was manipulated in Experiment 1 with tones that differed in intensity and in Experiment 2 with lifted cylinders that differed in weight. RESULTS: The results for Experiment 1 showed an interaction between task difficulty and hypoxia, indicating slowing of the preprocessing stage. Slowing was not found in Experiment 2. The absence of slowing in Experiment 2 is surprising and indicates that slowing may be confined to vision and audition and may not involve later, more central, stages. We discuss the need to measure cerebral oxygenation in order to understand the sharp differences between the the bottleneck hypothesis, developed by controlling SaO2, and the more traditional behavioral model which postulates multiple cognitive deficits. PMID- 9737762 TI - MR imaging of the central nervous system in divers. AB - BACKGROUND: Magnetic resonanse imaging (MRI) frequently reveals asymptomatic cerebral infarctions in the general population. HYPOTHESIS: The central nervous system (CNS) of divers is affected by a hyperbaric environment even if they are asymptomatic. METHODS: We examined 25 uniformed service divers by MRI and compared them with normal controls. RESULTS: Of 25 divers, 9 had CNS lesions vs. 2 of 25 controls (p = 0.02). There was a significant relationship between the CNS lesions, age, and smoking. CONCLUSION: The divers had a risk of accumulating CNS lesions. These results suggested that divers should undergo periodic medical evaluations and MRI brain scanning. PMID- 9737763 TI - Kinematic response of the neck to voluntary and involuntary flexion. AB - METHODS: The dynamic head-neck responses of human subjects and cadavers undergoing involuntary impact loading conditions have been studied extensively in order to define the kinematics of the neck undergoing rapid movements, but little detailed information is available regarding slower, voluntary motions. In this study, the dynamic kinematics of head/neck complex during subject-controlled, or voluntary head motion have been investigated to compare with the kinematics to involuntary response. Five male human subjects experienced two-types of posterior/anterior neck flexion: flexion initiated under their own volition, and flexion in response to -15Gx acceleration of the torso. Tri-axial photo target mounts were placed on a custom-fit plate at the mouth, on the first thoracic vertebra, and on the sled. High speed movie cameras captured the photo target motion. Linear and angular displacement and velocity of the head and T1 and sled were computed using customized data-processing software. The neck kinematics were represented by a 2-pin linkage which connected the anatomical origins of the head and T1. RESULTS: The results show that maximum neck flexion relative to the torso was not significantly different between the voluntary and involuntary head motions, but that the head motion was significantly greater during the involuntary sled maneuvers. Maximum flexion velocities of 450 and 1236 degrees x s(-1) were sustained during the voluntary and involuntary maneuvers, respectively. CONCLUSION: These findings are important in understanding the kinematics of the human head/neck complex undergoing rapid and slow movements, and will be valuable in future studies determining a realistic physiological performance corridor for the human neck. PMID- 9737764 TI - Further study on the carotid baroreflex system in the cardiovascular deconditioning induced by head-down tilt. AB - HYPOTHESIS: The responses of the carotid baroreflex and of the peripheral sympathetic system to stimulations induced by either lower body negative pressure (LBNP -40 mmHg) or cold pressor test were investigated in eight volunteers before and after 48 h in the -6 degrees head-down tilt (HDT). METHODS: Geometry (diastolic diameter and pulsatile distention) and dynamics (cross-sectional compliance and tangential tension) of the bulb and the common carotid artery were investigated using ultrasonic devices, echotracking and aplanation tonometry. The activity of the sympathetic system was evaluated through measurements of plasma concentrations of catecholamines (CAs) and 3,4-dihydroxyphenyl glycol (DHPG). RESULTS: During LBNP -40 mmHg, the pulsed tangential tension was decreased and the pressure amplification, induced by the reflexion of the pressure wave, was increased with no difference between before and after HDT. Since cross-sectional compliance and distensibility coefficient remained unchanged and the carotid contour of the waveform unaltered, it is concluded that the carotid reflexogenic area reads the same message during LBNP whether the cardiovascular system was deconditioned or not. Nonetheless, during LBNP after 48 h HDT, the heart rate accelerated faster and CAs and DHPG concentrations increased out of proportion, suggesting that the peripheral sympathetic activity was more reactive after HDT than before. Finally, forearm vascular resistances were measured in response to cold pressor test; they increased in the same proportion after HDT when compared with before. CONCLUSION: Results indicate that the carotid baroreflex and the peripheral sympathetic system were not deficient after 48 h HDT. PMID- 9737765 TI - Complex cataplexy. PMID- 9737766 TI - Telemedicine in Gulf War. PMID- 9737767 TI - Relativistic velocities and biological processes. PMID- 9737768 TI - Telomeres, hidden mosaicism, loss of heterozygosity, and complex genetic traits. AB - Telomeres appear to function as an endogenous timing mechanism in human beings. Telomere attrition not only provides a satisfactory explanation for some aspects of aging, it might also resolve enigmatic features of complex genetic traits that are age-dependent. If, with the passage of time, telomere attrition in human beings leads to genomic instability and particularly the loss of chromosomes, then the age dependency of phenotypic expressions of complex genetic traits might result from the temporal loss of heterozygosity and the consequent expression of disease-causing genes. In this way, telomere attrition might play a role not only in aging, but also in the diverse expression of complex genetic traits, such as essential hypertension, non-insulin-dependent diabetes mellitus, atherosclerosis, and cancer. PMID- 9737769 TI - Coping with complexity: lessons from the mathematical sciences. AB - I suggest five research paradigms fundamental in the mathematical sciences that seem relevant to the hunt for explanations of genetic susceptibility to complex diseases. I discuss a few ways in which these paradigms are, or are not, currently being followed in complex disease genetics. This essay is intended to provoke discussion and not to resolve any problems. It represents a written version of remarks that I made as "Discussant" at a session at the 1997 National Institutes of Health Research Festival. PMID- 9737770 TI - MTHFR association with arteriosclerotic vascular disease? AB - Complex diseases are far more common than diseases that follow simple Mendelian patterns of inheritance. Difficulties are experienced in the designing of experiments to dissect out the contribution of a single allele to a complex phenotype. We review the literature regarding a point mutation in methylenetetrahydrofolate reductase, a candidate gene for susceptibility to vascular diseases. PMID- 9737771 TI - A single mutation that results in an Asp to His substitution and partial exon skipping in a family with congenital contractural arachnodactyly. AB - Congenital contractural arachnodactyly (CCA) is an autosomal dominant disorder of connective tissue and is characterized by multiple congenital contractures, arachnodactyly, and external ear malformations. Recent investigations indicate that mutations in the fibrillin-2 gene (FBN2) cause CCA. Here, we report a G-->C transversion at nucleotide 3340 (G3340C) of FBN2 in a family with phenotypic characteristics of CCA. The G3340C mutation predicts the substitution of histidine for aspartic acid at amino acid residue 1114 (Asp1114His) and also alters the 5' donor splice site consensus sequence of exon 25. Reverse transcription/polymerase chain reaction and DNA sequence analyses demonstrate that this missense mutation also causes low level in-frame mis-splicing of exon 25 (del exon 25). Consequently, this single point mutation produces a heterogeneous population of mutant fibrillin-2 molecules in a single individual. Despite the complex manifestation of the mutation, it is associated with a relatively mild phenotype. Analysis of FBN2 allele expression in cultured dermal fibroblasts derived from the proband has shown that the mutant allele is preferentially expressed, contributing about 84% of the total transcript. This indicates that an overabundance of mutant transcript does not necessarily correlate with a more severe CCA phenotype. PMID- 9737772 TI - Histone H4 acetylation analyses in patients with polysomy X: implications for the mechanism of X inactivation. AB - In humans, it is thought that the X-inactivation phenomenon occurs no matter how many X chromosomes are present, and that only one of them remains active. Nevertheless, individuals who have an abnormal number of X chromosomes show a wide spectrum of abnormalities, which increase with the number of X chromosomes present in a given individual. It has been shown that the inactive X chromosome in female mammals is distinguished by a lack of histone H4 acetylation, and that this could be used as an accessible marker for distinguishing between Xi and Xa in spreads of metaphase chromosomes. We studied three X-polysomic patients for the presence of active chromatin by analysis of histone H4 acetylation on unfixed metaphase spreads. Using antisera to H4 acetylated at lysines 16, 8 and 5, respectively, we observed frequencies different from those expected from cells with only one underacetylated X chromosome. In particular, when antiserum to H4 acetylated at lysine 16 was used about 90% of the cells showed acetylation of all X chromosomes. This suggests a possible disturbance in the deacetylation process, probably due to the presence of multiple Xs. PMID- 9737773 TI - Familial aggregation of heart rate variability based on short recordings--the kibbutzim family study. AB - The objective of this study was to assess the familial aggregation of heart rate variability (HRV), a readily measurable noninvasive reflection of cardiac autonomic function. Familial correlations were analyzed in 451 kibbutz members aged 15-97 years belonging to 80 kindreds. Five-minute duration Holter recordings made during silent supine spontaneous breathing and metronomic breathing were analyzed in the time and frequency domains. The present analysis considers the familial correlations and the heritability estimates of two time-domain indices, the standard deviation (SD) of the R-R interval (RR), reflecting total variability, and the root mean square of successive differences in RR intervals (RMSSD), reflecting vagal (parasympathetic) tone. During free breathing, age- and sex-adjusted correlations between parents and their children (r=0.24 for both indices) and between adult siblings above 30 years of age (r=0.24 and t=0.34 for SD and RMSSD, respectively) were statistically significant, whereas spouse correlations (r=-0.04, r=-0.02 for SD and RMSSD, respectively) and correlations in younger siblings (r=-0.22 and r=0.01, respectively) were not. Significant heritability estimates were demonstrated for the two indices (h2=0.41 for SD and h2=0.39 for RMSSD). These findings suggest that familial aggregation of HRV characteristics is determined mostly by genetic factors and less so by environmental factors and provide a basis for continuing the investigation into the underlying genetic influences on HRV. PMID- 9737774 TI - No evidence of expansion of CAG or GAA repeats in schizophrenia families and monozygotic twins. AB - Many diseases caused by trinucleotide expansion exhibit increased severity and decreased age of onset (genetic anticipation) in successive generations. Apparent evidence of genetic anticipation in schizophrenia has led to a search for trinucleotide repeat expansions. We have used several techniques, including Southern blot hybridization, repeat expansion detection (RED) and locus-specific PCR to search for expanded CAG/CTG repeats in 12 families from the United Kingdom and 11 from Iceland that are multiplex for schizophrenia and demonstrate anticipation. The unstable DNA theory could also explain discordance of phenotype for schizophrenia in pairs of monozygotic twins, where the affected twin has a greater number of repeats than the unaffected twin. We used these techniques to look for evidence of different CAG/CTG repeat size in 27 pairs of monozygotic twins who are either concordant or discordant for schizophrenia. We have found no evidence of an increase in CAG/CTG repeat size for affected members in the families, or for the affected twins in the MZ twin sample. Southern hybridization and RED analysis were also performed for the twin and family samples to look for evidence of expansion of GAA/TTC repeats. However, no evidence of expansion was found in either sample. Whilst these results suggest that these repeats are not involved in the etiology of schizophrenia, the techniques used for detecting repeat expansions have limits to their sensitivity. The involvement of other trinucleotide repeats or other expandable repeat sequences cannot be ruled out. PMID- 9737775 TI - Elite endurance athletes and the ACE I allele--the role of genes in athletic performance. AB - Genetic markers that might contribute to the making of an elite athlete have not been identified. Potential candidate genes might be found in the renin angiotensin pathway, which plays a key role in the regulation of both cardiac and vascular physiology. In this study, DNA polymorphisms derived from the angiotensin converting enzyme (ACE), the angiotensin type 1 receptor (AT1) and the angiotensin type 2 receptor (AT2) were studied in 64 Australian national rowers. Compared with a normal population, the rowers had an excess of the ACE I allele (P<0.02) and the ACE II genotype (P=0.03). The ACE I allele is a genetic marker that might be associated with athletic excellence. It is proposed that the underlying mechanism relates to a healthier cardiovascular system. PMID- 9737776 TI - Microphthalmia with linear skin defects syndrome in a mosaic female infant with monosomy for the Xp22 region: molecular analysis of the Xp22 breakpoint and the X inactivation pattern. AB - This paper describes a female infant with microphthalmia with linear skin defects syndrome (MLS) and monosomy for the Xp22 region. Her clinical features included right microphthalmia and sclerocornea, left corneal opacity, linear red rash and scar-like skin lesion on the nose and cheeks, and absence of the corpus callosum. Cytogenetic studies revealed a 45,X[18]/46,X,r(X)(p22q21) [24]/46,X,del(X)(p22)[58] karyotype. Fluorescence in situ hybridization analysis showed that the ring X chromosome was positive for DXZ1 and XIST and negative for the Xp and Xq telomeric regions, whereas the deleted X chromosome was positive for DXZI, XIST, and the Xq telomeric region and negative for the Xp telomeric region. Microsatellite analysis for 19 loci at the X-differential region of Xp22 disclosed monosomy for Xp22 involving the critical region for the MLS gene, with the breakpoint between DXS1053 and DXS418. X-inactivation analysis for the methylation status of the PGK gene indicated the presence of inactive normal X chromosomes. The Xp22 deletion of our patient is the largest in MLS patients with molecularly defined Xp22 monosomy. Nevertheless, the result of X-inactivation analysis implies that the normal X chromosomes in the 46,X,del(X)(p22) cell lineage were more or less subject to X-inactivation, because normal X chromosomes in the 45,X and 46,X,r(X)(p22q21) cell lineages are unlikely to undergo X inactivation. This supports the notion that functional absence of the MLS gene caused by inactivation of the normal X chromosome plays a pivotal role in the development of MLS in patients with Xp22 monosomy. PMID- 9737777 TI - Negative association between asthma and variants of CC16(CC10) on chromosome 11q13 in British and Japanese populations. AB - The gene encoding Clara cell-derived inflammatory molecule CC16 has been cited as a candidate gene for atopic asthma on chromosome 1lq13. A genetic association study was performed with variants of the CC16 gene on chromosome 1lq13 in relation to asthma in British (n=275) and Japanese (n=300) populations. No significant association was found between asthma and CC16 genotypes, irrespective of atopic status in these two populations. These data suggest that CC16 might not be the major locus for asthma on 11q13. PMID- 9737778 TI - Genetic linkage of progressive pseudorheumatoid dysplasia to a 3-cM interval of chromosome 6q22. AB - Progressive pseudorheumatoid dysplasia (PPD), MIM 208230, is an autosomal recessive disorder, clinically characterized by spondyloepiphyseal dysplasia and progressive arthropathy. Linkage analysis of three families of different geographic and ethnic origin, including 11 affected individuals, showed strong evidence for localization of a gene for progressive pseudorheumatoid dysplasia to chromosome 6q with a maximum two-point lod score for D6S1647 of 8.34 at theta=0. Analysis of regions of homozygosity placed the gene in a 3-cM interval between D6S 1594 and D6S432. No significant shared haplotype was found for markers of the linked interval in the three families analyzed. Five genes encoding collagen and one encoding a specific procollagen-processing enzyme that map near this interval represent good candidates for the PPD gene. PMID- 9737779 TI - A missense Glu298Asp variant in the endothelial nitric oxide synthase gene is associated with coronary spasm in the Japanese. AB - Coronary spasm plays an important role in the pathogenesis of not only variant angina but also ischemic heart disease in general. However, the precise mechanism(s) by which coronary spasm occurs remains to be elucidated. Coronary spasm may arise from interactions between environmental and genetic factors. Endothelial-derived nitric oxide (NO) has been implicated in the control of vascular tone. We have recently shown that both basal and acetylcholine (ACh) induced NO activities are impaired in the coronary arteries of patients with coronary spasm. The purpose of this study has been to elucidate the possible variants that occur in the coding region of the endothelial nitric oxide synthase (eNOS) gene and that may be associated with coronary spasm. After initial screening in the entire 26 coding regions of the eNOS gene, we found a missense Glu298Asp variant in exon 7 in patients with coronary spasm. We subsequently performed a larger scale study involving 113 patients with coronary spasm and 100 control subjects, who were all diagnosed by intracoronary injection of ACh. The analysis revealed a significant difference in the distribution of the variant between the coronary spasm group (21.2%) and control group (9.0%; P=0.014 for dominant effect). Thus, we have found the missense Glu298Asp variant in the eNOS gene by the analysis of its entire 26 coding regions. The variant is significantly associated with coronary spasm. PMID- 9737780 TI - Molecular and clinical study of 183 patients with conotruncal anomaly face syndrome. AB - To investigate molecular and clinical aspects of conotruncal anomaly face (CAF), we studied the correlation between deletion size and phenotype and the mode of inheritance in 183 conotruncal anomaly face syndrome (CAFS) patients. Hemizygosity for a region of 22ql1.2 was found in 180 (98%) of the patients with CAFS by fluorescence in situ hybridization (FISH) using the N25(D22S75) DiGeorge critical region (DGCR) probe. No hemizygosity was found in three (2%) of the patients with CAFS by FISH using nine DiGeorge critical region probes and a SD1OP1 probe (DGA II locus). None of these three patients had mental retardation and just one had nasal intonation, which was observed in almost all of the 180 CAFS patients who carried deletions (mental retardation, 92%; nasal voice, 88%). Nineteen of 143 families (13%) had familial CAFS and 16 affected parents (84%) were mothers. Although only two of the affected parents had cardiovascular anomalies, the deletion size in the 16 affected parents and their affected family members, who were studied by FISH analysis, was the same. It indicates that extragenic factors may play a role in the genesis of phenotypic variability, especially in patients with cardiovascular anomalies. No familial cases were found among CAFS patients with absent thymus/DiGeorge anomaly (DGA). Also, in all 18 CAFS patients with completely absent thymus/DGA and all 6 CAFS patients with schizophrenia, it was revealed that the deletion was longer distally. A study of the origin of the deletion using microsatellite analyses in 48 de novo patients showed that in 65% of CAFS patients it was maternal, while in 64% of DGA patients it was paternal. The findings of this study indicated that CAF was almost always associated with the deletion of 22ql1.2. As well as the major features of the syndrome, other notable extracardiac anomalies were found to be susceptibility to infection, schizophrenia, atrophy or dysmorphism of the brain, thrombocytopenia, short stature, facial palsy, anal atresia, and mild limb abnormalities. PMID- 9737781 TI - Direct evidence of autosomal recessive inheritance of Arg24 to termination codon in purine nucleoside phosphorylase gene in a family with a severe combined immunodeficiency patient. AB - Purine nucleoside phosphorylase (PNP) deficiency is a rare immunodeficiency disease involving a T-lymphocyte-dysfunction that is fatal unless bone marrow transplantation is successful. In this study we undertook genetic analysis of a patient with PNP deficiency. Sequencing of the PNP gene, which is located on chromosome 14ql3, of the patient led to the identification of three point mutations in exon 2 at amino acid positions 20 (His, silent mutation), 24 (Arg- >termination codon) and 51 (Ser-->Gly). Intrafamilial sequence analysis of exon 2 revealed that both parents were heterozygous for the Arg24 and termination codon 24 alleles. Two of their three children had inherited different homozygous alleles, termination codon 24 for the patient, and Arg24 for his healthy sibling. Transcriptional termination was suggested as the mechanism giving rise to the disorder in this case. A lack of PNP protein was also confirmed by immunoblot analysis of the patient's hemolysate. This could be the first report providing evidence of autosomal recessive inheritance in PNP deficiency by sequence-based analysis. PMID- 9737783 TI - The 3' breakpoint of the yunnanese (Agammadeltabeta)0-thalassemia deletion lies in an L1 family sequence: implications for the mechanism of deletion and the reactivation of the Ggamma-globin gene. AB - We have cloned the junction region of the previously characterized Yunnanese (Agammadeltabeta)0-thalassemia deletion and the normal DNA region surrounding its 3' breakpoint. The sequence of 1138 bp of the 3'-flanking region and 555 bp of the normal region surrounding the 3' breakpoint was determined. The 5' breakpoint of the deletion is positioned between 116 and 117 bp upstream of the Agamma globin gene, and the 3' breakpoint at 34 bp downstream of the BglII site that lies approximately 12.7 kb upstream of the 3' deletion breakpoint of Chinese (Agammadeltabeta)0-thalassemia. There is no significant homology between the normal DNAs flanking the 5' and 3' breakpoints, except 2-bp (TG) identity and two pairs of short direct repeats at the deletion junction. The 3' breakpoint is within an L1 family sequence that normally occurs about 66 kb downstream of the beta-globin gene in 11p15. This L1 element is partially in reversed orientation at the 5' side and is therefore, presumably, an inactive element. This type of junction indicates illegitimate DNA breakage and end-joining involving the L1 sequence. Similar to many other deletions in the beta-globin locus, the Yunnanese (Agammadeltabeta)0-thalassemia deletion lacks a sequence characteristic of defined recombination factors near the deletion junction, suggesting that chromatin configuration and other locus-specific features are important. Computer aided analysis revealed that several transcriptional factors could bind to putative motifs present in the 3'-flanking sequence. It is possible that binding features of the distal L1 element are required to generate a suitable chromatin configuration for the recombination event in Yunnanese (Agammadeltabeta)0 thalassemia. This may also be related to the reactivation of the Ggamma-globin gene in adults with the Yunnanese (Agammadeltabeta)0-thalassemia mutation. PMID- 9737782 TI - Polymorphic tetranucleotide repeat site within intron 7 of the beta-amyloid precursor protein gene and its lack of association with Alzheimer's disease. AB - Mutations found in the beta-amyloid precursor protein (APP) gene in a small subset of patients with Alzheimer's disease (AD) are associated with the development of the disease. Several lines of evidence indicate that specific isoforms of APP generated by alternative splicing of the primary transcript may contribute to the etiology of AD. One of the isoforms, APP695, lacks the Kunitz protease inhibitor (KPI) domain and is produced predominantly in neurons by skipping exon 7 of the APP gene. Previous studies imply that the controlling elements for exon 7 skipping exist in the flanking sequences of the exon. Therefore, we have sequenced the human intron 7 of the APP gene and found a polymorphic tetranucleotide (ATTT)n repeat site within the intron 7. In 183 genetically unrelated subjects (97 AD patients and 86 controls), we found four alleles by polymerase chain reaction (PCR) amplification of the repeat site. Although no particular alleles are associated with AD, this newly identified polymorphic site may be useful in other genetic analyses since preliminary evidence suggests allele frequency differences between African Americans and Caucasians. PMID- 9737784 TI - The H-cadherin (CDH13) gene is inactivated in human lung cancer. AB - We have previously reported frequent allelic loss in chromosome bands 16q24.1 q24.2 in human lung cancer. Since the H-cadherin (CDH13) gene has been isolated and mapped to this common region of allelic loss, we have investigated this gene in human lung cancer. The reverse transcription/polymerase chain reaction technique has revealed the loss of expression in four (57%) of seven lung cancer cell lines. To study the CDH13 gene further, we have analyzed deletions, genetic alterations, and methylation status at the 5' region of this gene. Three (75%) of four cell lines that have lost expression show a deletion of the CDH13 locus accompanied by hypermethylation of the remaining allele. Moreover, hypermethylation has been observed in nine (45%) of 20 primary lung cancers. These results suggest that a combination of deletion and hypermethylation causes inactivation of the CDH13 gene in a considerable number of human lung cancers. PMID- 9737785 TI - Mutation of the start codon in the FRDA1 gene: linkage analysis of three pedigrees with the ATG to ATT transversion points to a unique common ancestor. AB - Friedreich ataxia (FRDA) is a progressive neurodegenerative disorder caused by loss-of-function mutations in the gene encoding frataxin. Most patients with FRDA have trinucleotide repeat expansions in both alleles of the FRDA 1 gene. In patients heterozygous for the expansion the second allele may be inactivated by a point mutation. We identified the ATG-->ATT (M11) mutation of the start codon in three independent families. Individuals with symptoms of FRDA in these families are compound heterozygous for the repeat expansion and the ATG mutation. To look for a common founder of the M11 mutation, a detailed linkage analysis employing six polymorphic chromosome 9 markers was performed. We found complete haplotype identity for two of the three chromosomes with the point mutation. The third case shows partial conformity and may be the result of a single recombination event. PMID- 9737786 TI - Identification, localization and characterization of the human gamma-synuclein gene. AB - We have identified and characterized a new member of the human synuclein gene family, gamma-synuclein (SNCG). This gene is composed of five exons, which encode a 127 amino acid protein that is highly homologous to alpha-synuclein, which is mutated in some Parkinson's disease families, and to beta-synuclein. The gamma synuclein gene is localized to chromosome 10q23 and is principally expressed in the brain, particularly in the substantia nigra. We have determined its genomic sequence, and established conditions for sequence analysis of each of the exons. The gamma-synuclein gene, also known as BCSG1, was recently found to be overexpressed in advanced infiltrating carcinoma of the breast. Our survey of the EST database indicated that it might also be overexpressed in an ovarian tumor. PMID- 9737787 TI - Molecular pathology of ovarian carcinomas. AB - There is evidence that ovarian cancer may be derived from the progressive transformation of benign and/or borderline tumours. Mutations involving different oncogenes and tumour suppressor genes accumulate during the process of malignant transformation, and the alterations of genes involved in the pathogenesis of familial ovarian cancer are probably early events in ovarian tumorigenesis. BRCA 1 and BRCA-2 act as classical tumour suppressor genes in hereditary tumours, but their role in sporadic tumours remains controversial; however, a high frequency of allele losses in BRCA-1 (17q) and BRCA-2 (13q) loci has been observed in both familial and sporadic tumours. The possible role of mismatch repair genes and microsatellite instability is also controversial, but a role for them has been proposed in borderline tumours. Mutations in K-ras are specific for mucinous tumours and may be related to mucinous differentiation. Finally, a role in tumour progression has been proposed for both c-erb B-2 and p53, but their practical value in prognosis remains questionable. PMID- 9737788 TI - ILK (beta1-integrin-linked protein kinase): a novel immunohistochemical marker for Ewing's sarcoma and primitive neuroectodermal tumour. AB - ILK (beta1-integrin-linked protein kinase) is a recently identified 59-kDa serine/threonine protein kinase that interacts with the cytoplasmic domain of the beta1-integrin containing four ankyrin-like repeats. We have developed a polyclonal antibody against ILK and explored the ILK immunoreactivity in normal human cells and tissues. ILK was mainly expressed in cardiac muscle and skeletal muscles. Surprisingly, ILK expression was observed in Ewing's sarcoma (ES; 100%), primitive neuroectodermal tumour (PNET; 100%), medulloblastoma (100%), and neuroblastoma (33.3%), whereas other small round cell sarcomas were not stained by the anti-ILK antibody. These results suggest that ILK could be a novel marker for tumours with primitive neural differentiation. Our findings support the notion that ES is a tumour that is closely related to PNET and that both originate from the neuroectoderm. ILK may be a sensitive and specific immunohistochemical marker and useful for the positive identification of ES and PNET in formalin-fixed, paraffin-embedded tissue sections. PMID- 9737789 TI - Biological and prognostic significance of stratified epithelial cytokeratins in infiltrating ductal breast carcinomas. AB - The biological significance of the differential expression of cytokeratin (CK) polypeptides in breast carcinomas is unclear. We examined the CK profiles of 101 primary infiltrating ductal breast carcinomas using monoclonal antibodies directed against 11 different CKs and against vimentin. Two major CK phenotypes were distinguished: first, a phenotype expressing only the simple-epithelial CKs 7 (variably), 8, 18 and 19, and secondly, a bimodal phenotype co-expressing significant amounts of one or more of the stratified-epithelial CKs 4, 14 and 17. The vast majority of G1 and G2 carcinomas had the simple-epithelium phenotype, as did a subgroup of G3 carcinomas. Interestingly, the majority (62%) of G3 carcinomas exhibited the bimodal phenotype, with the expression of CKs 4, 14 and 17 being statistically correlated with poor histological differentiation and absence of steroid hormone receptors. The distribution of vimentin only partially overlapped with that of these stratified-epithelial CKs. Prognostic analyses suggested that the presence of CKs 4, 14 and/or 17 was associated with short overall and disease-free survival in subgroups comprising G3, oestrogen-receptor negative and vimentin-negative tumours. In node-positive tumours the correlation between these CKs and a shorter disease-free interval attained statistical significance (log rank, 0.0096). Thus, abnormal CK profiles in ductal breast carcinomas appear to reflect disturbed regulation of differentiation-related gene expression programmes and may prove to be of clinical value. PMID- 9737790 TI - Apoptosis of cytotoxic T-cells in histiocytic necrotizing lymphadenitis. AB - Cell death is necrotic or apoptotic. In histiocytic necrotizing lymphadenitis (HNL), apoptosis is the main form of cell death, resulting in the creation of nuclear debris that is one of the characteristic features of HNL. To investigate the cell type of apoptotic cells, 12 cases of HNL were analyzed using the immunohistochemical staining for TIA-1, a cytotoxic granule of either cytotoxic T or NK cells. One quarter to over half of all apoptotic cells were positive for TIA-1, and some of the nuclear debris was also positive. The necrotic lesions of HNL were found to consist of nuclear debris, apoptotic cells, histiocytes and lymphocytes. The lymphocytes were mainly CD8-positive T-cells or CD4-positive cells, while B- and NK cells were only rarely observed. The number of TIA-1 positive lymphocytes was more closely related to the number of CD8-positive cells than to the number of CD4 cells. In double staining, the TIA-1 positive lymphocytes were mainly CD8 positive, but rarely CD4 positive. In HNL, then, CD8 positive cytotoxic T-cells are likely to undergo apoptosis. PMID- 9737791 TI - Multivariate karyometric approach in differential diagnosis of follicular thyroid neoplasms. A study of 31 cases. AB - A retrospective analysis of 19 follicular adenomas, 12 minimally invasive follicular carcinomas and 3 widely invasive follicular carcinomas of the thyroid was performed on 5-microm-thick Feulgen-stained paraffin sections by means of a semiautomatic system for picture analysis. The major aim was to assess the potential of multiparameter karyometry for separation of the first two tumour types. Sixteen planimetric and densitometric features were defined in each case on 200-300 randomly selected nuclei and processed by a number of uni- and multivariate statistical methods. Despite predominantly significant ANOVA results a substantial overlap between tumour groups limited the practical usefulness of any karyometric feature alone. Factor and cluster analyses indicated independence of planimetric and densitometric parameters from each other, which was of crucial importance in finding an optimal subset of variables for discriminant analysis. The classification rule derived from the latter procedure was checked by the "jack-knife" method, by classification of 3 widely invasive cancers and by hierarchical tumour clustering. Sensitivity and specificity of the model for detection of malignancy were 100% and 94.7%, respectively. A multivariate karyometric approach, when applied correctly, can be a useful tool for differentiation between follicular adenomas and minimally invasive follicular carcinomas of the thyroid. PMID- 9737792 TI - Vascular architecture of human urinary bladder carcinoma: a SEM study of corrosion casts. AB - The vascular architecture of five advanced invasive papillary tumours of the urinary bladder was investigated using corrosion casting and scanning electron microscopy. The superficial vasculature was composed predominantly of capillary systems of two types: dense flat networks with numerous interconnections and tightly packed tortuous loops, forming multiple irregular folds that reflected the papillary morphology of the tumours. The capillaries were supplied and drained by numerous straight nonanastomosing arterioles and venules, which arose by way of multiple branching of larger vessels originating from the mucosal plexus of the bladder. Differences between the tumours in the spatial arrangement of these vessels probably reflect different growth dynamics. The intramural parts of the tumours contained a chaotic network of straight, uniform capillaries with numerous sprouts, which was very different from the superficial capillary system. It is postulated that different angiogenesis-targeted growth factors may be expressed in the phases of exophytic growth and muscularis invasion of the tumour, leading to the formation of different microvascular patterns. PMID- 9737794 TI - Ultrastructural and immunohistochemical analysis of biopsy-proven chronic active mycocarditis with numerous clusters of lymphocytes. AB - A 60-year-old women was admitted to our hospital with deteriorating congestive heart failure. Although the diagnosis of active myocarditis was confirmed by right ventricular endomyocardial biopsy, this patient died of refractory heart failure during corticosteroid treatment. Numerous lymphocytic clusters were observed microscopically in the heart at autopsy. Most of the infiltrating cells in the clusters were positive for CD 8, HAM 56 or MHC class 2 antigen; few cells were positive for CD 56. Expression of perforin was found in some of the infiltrating cells. Electron microscopic examination revealed small lymphocytes adhering to the surface of injured cardiac myocytes. Close contact of these lymphocytes to macrophages was shown in the clusters. ICAM-1 and MHC class 1 antigens were strongly expressed in the cardiac tissue. These results indicate that cytotoxic T lymphocyte-mediated cytotoxicity had continued to operate during immunosuppressive therapy. Corticosteroids may not be suitable for the treatment of chronic active myocarditis when persistent expression of ICAM-1 is observed. PMID- 9737793 TI - Immunohistochemical localization of metallothionein in synovial tissue of patients with chronic inflammatory and degenerative joint disease. AB - Metallothioneins (MTs) are low-molecular-weight cytosolic proteins, which are thought to participate in metal homeostasis and protection against metal toxicity and oxidative stress. MT synthesis can be induced by a variety of inflammatory mediators and antirheumatic drugs, and high levels of MT have been implicated in resistance of cells to some antirheumatic drugs. We studied the expression and localization of MT in synovial tissue samples from patients with rheumatoid arthritis, ankylosing spondylitis, psoriatic arthritis or osteoarthritis (OA) by quantitative immunohistochemistry. Immunostaining for MT was detected in a large number of intimal lining cells in most of the investigated synovial tissue samples (75%). In a smaller proportion of samples (42%), some of the fibroblast like cells of the subsynovial layer were also MT positive. Immunostaining and double-staining experiments with antibodies against monocyte-, macrophage- and leucocyte-associated antigens suggested that most of the MT-positive cells were intimal fibroblast-like cells and subsynovial fibroblasts. However, there were no statistically significant differences in the intensity of staining for MT between the rheumatic diseases and OA at the single-cell level. Thus, MT is expressed in synovial tissue and may participate in homeostatic and protective functions. The interindividual variability in the expression of MT in synovial tissue may be related to the therapeutic efficacy of the gold compounds and chemotherapeutic antirheumatic drugs sequestered by MT. PMID- 9737795 TI - Numbers of nuclei in different tissue compartments of fetal ventricular myocardium from 16 to 35 weeks of gestation. AB - The aim of this study was to examine mechanisms of growth in different tissue compartments of the ventricular myocardium of prenatal human hearts. To this end, stereological methods were applied in order to estimate tissue volumes and total numbers of myocyte, connective tissue and endothelial nuclei in hearts collected after death at between 16 and 35 weeks of gestation. Volumes of tissue compartments were obtained after multiplying volume densities (estimated by test point counting) by ventricular volumes (estimated from ventricular mass and tissue density). Absolute numbers of nuclei were calculated in similar fashion from corresponding nuclear packing densities (estimated using physical disectors). The volumes of all three tissue compartments increased linearly over the period of gestation examined, and in each case, the increase in tissue volume appeared to be due entirely to proliferation. Numbers of all three types of nuclei increased linearly whilst tissue volumes per nucleus remained constant. The net rate of production of myocyte nuclei was 35 x 10(7) per week (2.1 million nuclei per hour). The net rate of production of connective tissue nuclei was 12 x 10(7) per week (0.7 million nuclei per hour) and that for endothelial cell nuclei was 5.1 x 10(7) per week (0.3 million nuclei per hour). Predictions are made about the postnatal ages at which adult ratios of different nuclear types might be attained. PMID- 9737796 TI - Morphological and functional alterations to sinusoidal endothelial cells in the early phase of endotoxin-induced liver failure after partial hepatectomy in rats. AB - Liver failure following major hepatectomy is characterized pathologically by massive hepatic necrosis, which is thought to begin with injury of sinusoidal endothelial cells (SECs). To examine the early events of SECs leading to hepatic damage, we performed time-course analyses of the morphological and functional perturbation of SECs after endotoxin administration to hepatectomized rats. At 1.5 h after endotoxin injection, when hepatocellular damage was not yet evident, SECs showed augmented expression of intercellular adhesion molecule-1, with frequent adherence of infiltrating leucocytes and ultrastructural features of defenestration and hypertrophied cytoplasm enriched with cell organelles. The serum level of hyaluronate, as an indicator of the functional state of SECs, was significantly elevated. At 3 h, SECs underwent necrosis and disruption, accompanied by fibrin deposits with concomitant hepatocellular necrosis. The morphological and functional alterations of SECs precede necrotic changes in hepatocytes and SECs in endotoxin-induced liver failure after partial hepatectomy. PMID- 9737797 TI - Mesangial cell activation in the collagenofibrotic glomerulonephropathy. Case report and review of the literature. AB - Collagenofibrotic glomerulonephropathy is a new disease entity of unknown pathogenesis, which is characterized by the deposition of type III collagen within the mesangial matrix. We have investigated a case in which many mesangial cells in the type III collagen-deposited glomeruli were alpha-smooth muscle actin (ASMA) positive and showed an increase of subplasmalemmal filaments, indicating the activation and myofibroblastic transformation. It is suggested that the activated mesangial cells may synthesize the type III collagen deposited in the subendothelial space and mesangial matrix. PMID- 9737798 TI - Synovial chondromatosis: clonal chromosome changes provide further evidence for a neoplastic disorder. AB - Synovial chondromatosis is a rare lesion, which is still believed by most authors to be reactive rather than neoplastic. We report on a case of synovial chondromatosis with clonal chromosomal changes [43,XX,der (1) t (1;13) (p21 22;q21),-6,-13,-14, add(21) (q21)]. The presence of clonal chromosomal changes in this and in three previously reported cases suggests that synovial chondromatosis is a true neoplastic lesion. PMID- 9737799 TI - Screening for diabetic retinopathy in the 21st century. PMID- 9737800 TI - Autoimmunity in diabetic autonomic neuropathy: does the immune system get on your nerves? AB - Symptomatic autonomic neuropathy is a devastating occasional complication of diabetes mellitus, especially Type 1. Although the full-blown clinical syndrome is not common, dysfunction of the autonomic nerves is detectable in up to 40% of Type 1 diabetic patients but its aetiopathogenesis is poorly understood. There is evidence to suggest that the damage to the autonomic nerves may be immune mediated. This evidence is reviewed in the following article. PMID- 9737801 TI - Cow's milk consumption, disease-associated autoantibodies and type 1 diabetes mellitus: a follow-up study in siblings of diabetic children. Childhood Diabetes in Finland Study Group. AB - Evidence from case-control studies for the diabetogenicity of introduction of cow's milk-based formulas at early age in infancy is inconclusive. We followed siblings of children with Type 1 diabetes mellitus (Type 1 DM) to investigate a possible relationship between cow's milk consumption during infancy or later in childhood and the emergence of diabetes-associated autoantibodies and progression to clinical Type 1 DM. A cohort of 725 initially unaffected 0 to 25-year-old siblings of 801 index children with Type 1 DM diagnosed in 1986-1989 participated in the study (82% of those invited). The siblings were observed for Type 1 DM associated autoantibodies at intervals of 3-12 months for 4 years, starting from the diagnosis of Type 1 DM in the index child. The follow-up for Type 1 DM started at the same time and ended on 31 October 1995. The combined prevalence of Type 1 DM associated autoantibodies (islet cell antibodies (ICA), insulin autoantibodies (IAA), GAD autoantibodies (GADA), and/or antibodies to the insulinoma associated cDNA2 protein (IA-2A)) was 13.6% (95/697) at the beginning of the study. Of the initially seronegative siblings, 7.5% (45/602) converted to antibody positivity during 4 years, and of all siblings 4.6% (33/725) developed Type 1 DM during the total follow-up time. The age at introduction of supplementary milk feeding was not significantly related to seroconversion to positivity for Type 1 DM associated autoantibodies or to the development of Type 1 DM in the siblings. When adjusted for age, sex, infant feeding patterns, and maternal age and education, high milk consumption in childhood (> or =3 glasses daily) was associated with more frequent emergence of Type 1 DM-associated autoantibodies than low consumption (<3 glasses daily) (adjusted odds ratio 3.97, 95% confidence interval 1.3-11.7, p = 0.01). There was a non-significant association between high milk consumption and progression to clinical Type 1 DM (adjusted hazard ratio 2.75, 95% confidence interval 0.9-8.4, p = 0.07). To conclude, this study suggests that high consumption of cow's milk during childhood may be associated both with seroconversion to positivity for diabetes associated autoantibodies and progression to clinical Type 1 DM among siblings of children with diabetes. PMID- 9737802 TI - Effect of pancreas transplantation and immunosuppression on proinsulin secretion. AB - Insulin resistance and increased demand for insulin secretion occur after successful pancreas transplantation. To investigate the potential effects of immunosuppression and pancreas transplantation on fasting beta-cell function, we studied fasting proinsulin and 32,33 split proinsulin secretion cross-sectionally and longitudinally in segmental pancreatic graft recipients (SPx, n = 18); in whole-pancreas graft recipients (WPx, n = 13); in nondiabetic kidney transplant recipients (Kx, n = 14) and in normal subjects (Ns, n = 14). Basal insulin secretion rates were significantly increased in SPx 15.8 (1.7), WPx 24.4 (4.5) and Kx 22.1 (2.1) vs Ns 9.7 (1.6) pmol min(-1) l(-1), p < 0.05, mean (SEM). Total proinsulin, intact proinsulin and 32,33 split proinsulin concentrations were significantly higher in all the transplanted groups than in normal subjects (p < 0.05), whereas the total proinsulin to C-peptide ratio and the 32,33 split proinsulin ratio were higher in SPx than in WPx, Kx and Ns (< 0.05). In the longitudinal study, beta-cell function in terms of proinsulin secretion remained stable for 1 year. In conclusion, fasting glucose homeostasis in pancreas-kidney transplant recipients is obtained at the expense of increased proinsulin secretion and increased insulin secretion rates, primarily induced by immunosuppression. In segmental pancreas graft recipients, increased fasting proinsulin and 32,33 split proinsulin relative to the number of beta-cells transplanted indicate more stress on the residual beta-cell and therefore higher secretory demand than in whole pancreas transplant recipients. PMID- 9737803 TI - Intramuscular versus subcutaneous injection of soluble and lispro insulin: comparison of metabolic effects in healthy subjects. AB - The aim of this study was to compare the glucodynamic effects of soluble insulin and the rapid acting insulin analogue insulin lispro after subcutaneous (s.c.) and intramuscular (i.m.) injection. Twelve healthy male volunteers (age 26.8 +/- 1.7 years, BMI 23.2 +/- 2.3 kg m(-2); mean +/- SD) participated in this single centre, open-labelled, euglycaemic glucose clamp study on four different days. Soluble insulin or insulin lispro (0.2 U kg(-1)) were injected s.c. or i.m. into the thigh by syringe. The glucodynamic effects were assessed by registering the glucose infusion rates necessary to maintain blood glucose at 5.0 mmol l(-1) for the subsequent 420 min. Intramuscular injection of soluble insulin led to an earlier peak of metabolic action when compared to s.c. administered soluble insulin (tmax 138 +/- 29 vs 179 +/- 34 min; p < 0.05). The maximal metabolic effect and metabolic activity during the first 2 h after i.m. and s.c. injection of soluble insulin were comparable (GIRmax 9.7 +/- 2.3 vs 7.8 +/- 2.3 mg kg(-1) min(-1); n.s., AUC0-120min 0.60 +/- 0.18 vs 0.50 +/- 0.15 g kg(-1) 120 min; n.s.). Subcutaneous administration of insulin lispro led to a metabolic effect resembling that induced by i.m. application of soluble insulin (tmax 116 +/- 26 vs 138 +/- 29 min; n.s., GIRmax 11.1 +/- 2.3 mg vs 9.7 +/- mg kg(-1) min(-1); n.s.). However, the overall metabolic response during the first 2 h after injection was higher with s.c. insulin lispro (AUC0-120min 0.81 +/- 0.26 vs 0.60 +/- 0.18 g kg(-1) 120 min; p < 0.05). The glucodynamic activity of i.m. applied insulin lispro was comparable to that of lispro s.c.. Following i.m. injection of soluble insulin, the metabolic activity peaked more rapidly than with s.c. administration. In contrast, the metabolic effect of insulin lispro was similar with either route. The time-action profile of i.m. injected soluble insulin lies between that of s.c. applied soluble insulin and insulin lispro. PMID- 9737805 TI - Problem of amputations in patients with newly diagnosed diabetes mellitus. AB - A reduction of 50% or more in diabetes-related amputations is a primary target of the St Vincent Declaration. This is thought to be achievable because both primary and secondary preventative healthcare strategies are effective in reducing the incidence of diabetic foot ulceration and progression to amputation. Unfortunately there is a group who cannot benefit from preventative health care, that is, newly diagnosed diabetic patients with already established severe complications. Using our population-based district diabetes information system we investigated, during the period 1 January 1992 to 31 December 96, the incidence and prevalence of lower extremity amputations (LEAs) and the proportion occurring in patients newly or recently diagnosed as having diabetes. Seventy-nine diabetic patients (59 male, 20 female) were recorded as having had 94 LEAs, the incidence of diabetes-related LEA being 475 per 100,000 diabetic patient-years. Of these LEAs 16 (20.2%) were performed within 1 year of diabetes being diagnosed. This study highlights an appreciable and previously unrecognized problem: patients presenting with established complications of diabetes who cannot benefit from secondary preventative healthcare. These patients pose a potential obstacle to achieving targets for reductions in diabetes-related amputations. PMID- 9737804 TI - Insulin management and metabolic control of type 1 diabetes mellitus in childhood and adolescence in 18 countries. Hvidore Study Group on Childhood Diabetes. AB - Insulin regimens and metabolic control in children and adolescents with Type 1 diabetes mellitus were evaluated in a cross-sectional, non-population-based investigation, involving 22 paediatric departments, from 18 countries in Europe, Japan, and North America. Blood samples and information were collected from 2873 children from March to August 1995. HbA1c was determined once and analysed centrally (normal range 4.4-6.3%, mean 5.4%). Year of birth, sex, duration of diabetes, height, body weight, number of daily insulin injections, types and doses of insulin were recorded. Average HbA1c in children under 11 years was 8.3 +/- 1.3% (mean +/- SD) compared with 8.9 +/- 1.8% in those aged 12-18 years. The average insulin dose per kg body weight was almost constant (0.65 U kg(-1) 24 h( 1)) in children aged 2-9 years for both sexes, but there was a sharp increase during the pubertal years, particularly in girls. The increase in BMI of children with diabetes was much faster during adolescence compared to healthy children, especially in females. Sixty per cent of the children (n = 1707) used two daily insulin injections while 37% (n = 1071) used three or more. Of those on two or three injections daily, 37% used pre-mixed insulins, either alone or in combination with short- and intermediate-acting insulin. Pre-adolescent children on pre-mixed insulin showed similar HbA1c levels to those on a combination of short- and long-acting insulins, whereas in adolescents significantly better HbA1c values were achieved with individual combinations. Very young children were treated with a higher proportion of long-acting insulin. Among adolescent boys, lower HbA1c was related to use of more short-acting insulin. This association was not found in girls. We conclude that numerous insulin injection regimens are currently used in paediatric diabetes centres around the world, with an increasing tendency towards intensive diabetes management, particularly in older adolescents. Nevertheless, the goal of near normoglycaemia is achieved in only a few. PMID- 9737806 TI - Outcome on diabetic foot complications in relation to clinical examination and quantitative sensory testing: a case-control study. AB - A total of 405 diabetic patients who first attended St Thomas' Diabetes Clinic between 1982 and 1985 had a detailed standardized computerized first visit record, including a structured foot examination and toe vibration perception thresholds (VPT, Biothesiometer), were reviewed in 1995. None of the patients had a history of foot ulceration at first visit. Twenty-five patients (6.2%) developed foot ulcers (n = 11, 2.7%) or had an amputation (n = 14, 3.5%) over a mean 12-year period. Twenty of these patients were then individually matched with 3 non-ulcer patients. Statistically significant odds ratios (OR) were found for a baseline abnormal age-adjusted toe VPT (OR 4.38, CI 1.11-17.26; p = 0.01); abnormal clinical examination (at least 1 abnormality out of: ankle jerks, tuning fork or cotton wool sensation; OR 2.3, CI 1.00-5.20; p < 0.01); and HbA1 (OR 1.30, CI 1.01-1.66; P < 0.02) in patients who subsequently developed lower extremity complications. The sensitivity of VPT (70%) was better than that for clinical testing (55 %) in predicting long-term complications, although all tests showed similar specificity (70-72%). The risk of events also doubled for every 10 years of diabetes (OR 2.10, CI 1.11-4.30; p = 0.02). We conclude that age corrected VPT measurements, which are objective and simple to perform, are better predictors of future foot complications than semi-quantitative tests in diabetes clinics. We encourage their use in the campaign to reduce the morbidity of diabetic peripheral neuropathy. PMID- 9737808 TI - Survey of diabetic retinopathy screening services in England and Wales. AB - A postal survey of diabetologists was conducted regarding the provision of diabetic retinopathy screening services in England and Wales. About 2.5 million people had no existing or planned screening service. For the rest, the perceived percentage of patients with diabetes screened varied from less than 25% to more than 90%. Multiple modes of screening were used in most units. Lack of funding was identified as the major reason for non-provision of an adequate screening service. About 18% of the units had to use research or charitable funds for screening. Only 50% of the units using optometrists for screening had standard protocols for referral. The average wait before an ophthalmologist's opinion on sight threatening retinopathy detected by screening was unacceptably high in some units. We would suggest that establishment of identical screening protocols and provision of adequate funding on a national basis ought to be the priority if incidence of blindness from diabetic retinopathy is to be reduced according to the St Vincent Declaration. PMID- 9737807 TI - Troglitazone, an insulin action enhancer, improves glycaemic control and insulin sensitivity in elderly type 2 diabetic patients. AB - The management of Type 2 diabetes mellitus with currently available oral agents may be complicated in the elderly by an increased frequency of side-effects. The effects of troglitazone, an insulin action enhancer, were studied in elderly patients with Type 2 diabetes in a double-blind, parallel-group, placebo controlled trial. A total of 229 patients (41% male), mean age 75 (range 69-85) years, with two fasting capillary blood glucose values > or =7 and < or =15 mmol l(-1) (and within 4.0 mmol l(-1) of each other) and previously treated with either diet alone (30%) or oral hypoglycaemic agents, were randomized to placebo or troglitazone 400 mg once daily or 200 mg twice daily, or 800 mg once daily or 400 mg twice daily, for 12 weeks. After 12 weeks' treatment, fasting serum glucose was significantly lower in troglitazone-treated patients (troglitazone, adjusted geometric mean 9.4-10.4 mmol l(-1) vs placebo 12.7 mmol l(-1), p < 0.001). Adjusted geometric mean fructosamine was also lower in troglitazone treated patients by 5 to 15% compared to placebo (P < 0.05 at all doses except 400 mg od). There was no significant difference between troglitazone doses for improvement in glycaemic control. Troglitazone lowered adjusted geometric mean fasting plasma insulin by 27-34% compared to placebo (P < 0.001) and insulin sensitivity (HOMA-S) improved by 9-15% in all troglitazone dose groups (p < 0.001). Troglitazone also lowered serum non-esterified fatty acids and triglyceride. Adverse event incidence in troglitazone-treated patients was similar to that in patients treated with placebo. No weight gain or symptomatic hypoglycaemia was recorded at any of the doses studied. Troglitazone is effective and well tolerated in elderly patients with Type 2 diabetes mellitus, providing improved glycaemic control in the absence of weight gain. PMID- 9737809 TI - Screening for diabetic retinopathy: the utility of nonmydriatic retinal photography in Egyptian adults. AB - Although regular screening for diabetic retinopathy with ophthalmoscopy or retinal photography is widely recommended in the United States and Europe, few reports of its use in developing countries are available. We compared the performance of screening by retinal photography with that of indirect ophthalmoscopy by using data from a population-based survey of diabetes and its complications in Egypt. During that project, 427 persons with diabetes underwent an eye examination and fundus photography with a non-mydriatic camera through a dilated pupil. Data from the examinations of the right eye of each patient are presented. Ninety-two (22%) of the 427 retinal photographs were ungradable; in 58 eyes (63%), this was due to media opacity (42 eyes with cataract, 3 with corneal opacity, and 13 with both). Agreement between retinal photography and indirect ophthalmoscopy was poor (kappa = 0.33; 95% CI = 0.27-0.39) and primarily due to the large number of eyes (n = 79) with ungradable photographs that could be graded by ophthalmoscopy. None of these eyes was judged by ophthalmoscopy to have sight-threatening retinopathy. Fifty-four photographs were diagnosed with greater retinopathy than found on ophthalmoscopy. Retinal photography with the nonmydriatic camera through a dilated pupil is a useful method to screen for diabetic retinopathy in most adults in Egypt. However, such screening strategies have limited use in older persons and in persons with corneal disease or cataract. PMID- 9737810 TI - High glucose uptake by adipocytes in a type 1 diabetic patient with a partial 'honeymoon' period. AB - Hypoglycaemic episodes and low insulin requirements are frequently seen in the early phase of treatment of Type 1 diabetes mellitus but the mechanism is not clear. We present a diabetic patient with recurrent hypoglycaemia in the early phase of insulin treatment. A very high glucose transport in adipocytes (basal: 176 and insulin stimulated glucose transport 10(-7) mol l(-1): 335 fl cell(-1) s( 1)) was found when compared with reference laboratory diabetic patients (basal: 59 +/- 10 and insulin 10(-7) mol l(-1): 106 +/- 7 fl cell(-1) s(-1), mean +/- SE) and with reference laboratory of non-diabetic subjects (basal: 106 +/- 6 and insulin 10(-7) mol l(-1): 188 +/- 15 fl cell(-1) s(-1)). Insulin binding to adipocytes was in the normal range. The patient was studied again 1 year later when the partial clinical remission had disappeared, and the glucose transport in adipocytes had decreased. In conclusion, an increase in glucose uptake by peripheral tissues may be among the mechanisms of the partial 'honeymoon' period of diabetic patients. PMID- 9737811 TI - Access to diabetes treatment in northern Ethiopia. AB - Treatments for diabetes in Ethiopia are at present only available in hospitals so many patients must travel great distances to obtain insulin, tablets, and diabetes education. We reviewed all 496 people with diabetes attending the diabetic clinic at Gondar Hospital (281 with Type 1 (insulin-dependent) diabetes mellitus (DM) and 215 with Type 2 (non-insulin-dependent) DM. Half of the patients came from rural areas, all but 3 of them travelling more than 20 km, one quarter of them more than 100 km and 33 patients (13%) more than 180 km. It is likely that many patients who fail to attend from the more distant areas have died. We are developing a scheme which would enable diabetic patients to be treated at rural health centres by nurses trained in the principles of diabetes care which could greatly improve the outlook for diabetic patients in Ethiopia. PMID- 9737812 TI - Antibodies to GAD in diabetic patients with chronic hepatitis C. PMID- 9737813 TI - A combined treatment for severe diabetic neuropathy symptoms. PMID- 9737814 TI - Parental hypertension and risk of diabetic nephropathy. PMID- 9737815 TI - Pharmacokinetics of intravenously and orally administered eprosartan in healthy males: absolute bioavailability and effect of food. AB - Eighteen healthy males received a single 300 mg oral dose of eprosartan as the commercial wet granulation formulation under fasting conditions and following a high-fat breakfast and a single 20 mg intravenous (i.v.) dose. The pharmacokinetics of i.v. eprosartan (mean +/- S.D.) were characterized by a low systemic plasma clearance (131.8 +/- 36.2 mL min(-1)) and a small steady-state volume of distribution (12.6 +/- 2.6 L). Oral bioavailability averaged 13.1%, due to incomplete absorption. In vitro dynamic flow cell dissolution data showed that pH-dependent aqueous solubility of eprosartan is one factor which limits absorption. Eprosartan terminal half-life was shorter after i.v. (approximately 2 h) versus oral (approximately 5-7 h) administration, which may be due to detection of an additional elimination phase or absorption rate-limited elimination following oral administration. Oral administration of eprosartan following a high-fat meal compared with fasting conditions resulted in a similar extent of absorption (based on AUC), but a decreased absorption rate. Cmax was approximately 25% lower, and a median delay of 1.25 h in time to Cmax was observed when eprosartan was administered with food. These minor changes in exposure are unlikely to be of clinical consequence; therefore, eprosartan may be administered without regard to meal times. PMID- 9737817 TI - Steady-state pharmacokinetics of diltiazem and hydrochlorothiazide administered alone and in combination. AB - Diltiazem and hydrochlorothiazide are widely used to treat cardiovascular disease, often in combination. The purpose of this investigation was to determine whether a drug-drug pharmacokinetic interaction exists between diltiazem and hydrochlorothiazide. In a randomized, crossover, open study, multiple doses of diltiazem (60 mg four times daily for 21 doses) and hydrochlorothiazide (25 mg twice daily for 11 doses) were administered alone and in combination on three separate occasions to 20 healthy male volunteers. Trough and serial blood samples were collected and plasma was assayed for diltiazem, hydrochlorothiazide, and diltiazem metabolites (desacetyldiltiazem and N-desmethyldiltiazem) using HPLC. Total urine was also collected and quantified for hydrochlorothiazide. Coadministered hydrochlorothiazide did not significantly (p > 0.05) alter diltiazem (alone versus combination) steady-state maximum plasma concentration (Css(max); 145 versus 158 ng mL(-1), respectively), time to maximum plasma concentration (t(max); 3.0 versus 2.8 h, respectively); area under the plasma concentration-time curve (AUCss; 688 versus 771 ng x h mL(-1)), oral clearance (Cl(oral); 96.2 versus 88.0 L h(-1)), or elimination half-life (t(1/2); 5.2 versus 5.2 h). Similarly, administration of diltiazem did not significantly (p > 0.05) influence hydrochlorothiazide (alone versus combination) Css(max) (221 versus 288 ng mL(-1)), t(max) (1.8 versus 2.0 h), AUCss (1194 versus 1247 ng x h mL(-1)), Cl(oral) (22.4 versus 21.2 L h(-1)); t(1/2) (9.8 versus 9.6 h), or renal Cl (15.5 versus 15.2 L h(-1)). In conclusion, a clinically significant pharmacokinetic interaction between diltiazem and hydrochlorothiazide does not exist. PMID- 9737816 TI - Pharmacokinetic and pharmacodynamic changes of furosemide after intravenous and oral administration to rats with alloxan-induced diabetes mellitus. AB - Because some physiological changes occurring in diabetes mellitus patients could alter the pharmacokinetics and pharmacodynamics of the drugs to treat the disease, the pharmacokinetics and pharmacodynamics of furosemide were investigated after intravenous (i.v.) and oral administration of the drug (6 mg per whole body weight) to control rats and alloxan-induced diabetes mellitus rats (AIDRs). After i.v. administration, the total body clearance (5.47 versus 7.05 mL min(-1) kg(-1)) was significantly slower in AIDRs and this was due to significantly slower renal clearance (2.35 versus 4.33 mL min(-1) kg(-1)) because the nonrenal clearance was comparable between two groups of rats. The 8 h urinary excretion of furosemide after i.v. administration decreased significantly (2280 versus 3760 microg) in AIDRs due to impaired kidney function; the glomerular filtration rate measured by creatinine clearance was significantly slower (2.86 versus 4.33 mL min(-1) kg(-1)) and both the plasma urea nitrogen (43.5 versus 17.3 mg dL(-1)) and kidney weight (0.953 versus 0.749% of body weight) increased significantly in AIDRs. This resulted in a significant decrease in the 8 h urine output per g kidney (17.8 versus 43.6 mL) in AIDRs. However, the 8 h diuretic efficiency was not significantly different between two groups of rats. After oral administration, the area under the plasma concentration-time curve from time 0 to 8 h decreased significantly in AIDRs (1200 versus 1910 microg x min mL(-1)) due to considerably decreased absorption of furosemide from gastrointestinal tract of AIDRs. After oral administration, the 8 h urine output per g kidney (18.6 versus 36.4 mL) also decreased significantly in the AIDRs due to significantly decreased 8 h urinary excretion of furosemide (405 versus 2210 microg), however, the 8 h diuretic efficiency increased significantly (127 versus 35.2 mL mg(-1)) in AIDRs. PMID- 9737818 TI - Blood disposition and urinary excretion kinetics of methazolamide following oral administration to human subjects. AB - The pharmacokinetic disposition of methazolamide (MTZ) was studied in five healthy volunteers following administration of a single oral dose. Drug concentrations in blood, plasma, and urine were measured by HPLC. Over the range of plasma concentrations observed in vivo, MTZ free fraction (measured by ultrafiltration) was 0.28. Being a carbonic anhydrase inhibitor, MTZ would be expected to distribute into, and be sequestered by, red blood cells. For this reason, MTZ disposition was characterized utilizing blood concentrations as the reference. Using a two-compartment model, a series of differential equations were simultaneously fitted to blood concentrations and urinary excretion data generating estimates for k10 (0.035 +/- 0.019 h(-1)), k12 (0.200 +/- 0.036 h( 1)), k21 (0.077 +/- 0.046 h(-1)), k(a) (0.304 +/- 0.064 h(-1)), Vc (1.1 +/- 0.18 L) and f(r) (fraction excreted renally, 0.61 +/- 0.14). Total blood clearance was 0.037 +/- 0.020 L h(-1). The model estimate of elimination half-life (126 +/- 61 h) was consistent with drug binding to a high affinity carbonic anhydrase isozyme in the erythocyte. Estimates of MTZ renal clearance and renal excretion ratio were 0.021 +/- 0.010 L h(-1) and 0.16 +/- 0.06, respectively. Overall, the prolonged elimination of MTZ from the blood is the result of extensive erythrocyte distribution and tubular reabsorption by the kidney. PMID- 9737819 TI - Evaluation of the effect of food on the pharmacokinetics of avitriptan. AB - Bioavailability of avitriptan was found to decrease significantly when administered 5 min after a standard high fat meal. The studies described herein were carried out to gain insight into the mechanism of this food effect. A series of studies were conducted in humans to assess the effect of timing of meal, type of meal, gastric pH, change in the formulation and dose on the bioavailability of avitriptan. Avitriptan was administered as a 50 mg capsule under fasted condition and at 30 min, 1, 2 and 4 h after a standard high fat meal. The reduction in avitriptan bioavailability persisted even at 4 h post high fat meal, although as the time interval between the meal and dose increased, the effect of meal tended to decrease. Bioavailability of avitriptan also decreased significantly when the drug was administered after a high protein and a high carbohydrate meal. Elevation in gastric pH caused by food was not found to be responsible for the food-related decrease in bioavailability of avitriptan since ranitidine pretreatment did not lead to a decrease in bioavailability. When administered as a 50 mg 14C-labeled solution after a standard high fat meal, bioavailability of avitriptan decreased although the decrease was less compared with that observed for a capsule dosage form. Plasma concentrations and cumulative urinary excretion of total radioactivity also decreased in the fed condition, indicating the absorption of avitriptan was affected. The decrease in avitriptan AUC was somewhat more pronounced than the decrease in the exposure to the total radioactivity suggesting a food-related increase in the first-pass metabolism of avitriptan. Effect of the standard high fat meal on avitriptan administered as a 150 mg capsule was similar to that observed at the 50 mg dose. Overall, the results indicate that bioavailability of avitriptan is significantly reduced irrespective of the type of meal, dose and dosage form and the effect persists for as long as 4 h post meal. Thus, it appears that avitriptan absorption and bioavailability are highly sensitive to presence of food in the stomach and any food-related changes in gastric emptying time and gastrointestinal motility. PMID- 9737820 TI - Uptake of verteporfin by articular tissues following systemic and intra-articular administration. AB - Photodynamic therapy (PDT) using the photosensitizer BPD-Verteporfin (liposomal benzoporphyrin derivative-monoacid ring A) has been shown in previous studies to be effective in the amelioration of inflammatory arthritis in both the MRL-lpr mouse and the New Zealand White (NZW) rabbit models, and could potentially offer alleviation of certain inflammation-related symptoms of rheumatoid arthritis. Time and dose dependency of BPD-MA tissue uptake was carried out in the inflamed synovium and other articular and peri-articular tissues following intravenous and intra-articular administration in the NZW rabbit model. As some articular and peri-articular tissues are difficult to extract, this study uses a rapid fluorimetric sampling of tissues following dissolution in Soluene 350. Our results showed that i.v. injected BPD-MA preferentially distributed in the inflamed synovium, and in tissues with a high degree of vascularization. Little or no association was found with avascular tissues such as cartilage and tendons. Clearance from the synovium was rapid, supporting earlier rather than late light treatment. Much higher association of BPD-MA with the synovium was achieved using intra-articular injection, and BPD-MA concentrations were maintained at relatively steady levels for several hours. These observations support the possibility that PDT could offer a safe, highly versatile clinical option for the management of inflamed joints in autoimmune disorders. PMID- 9737821 TI - Bioavailability of morphine after administration of a new bioadhesive buccal tablet. AB - A new bioadhesive buccal morphine tablet was developed for controlled release delivery of drug and improved bioavailability compared with oral controlled release tablet. In order to characterize the pharmacokinetic properties of this bioadhesive buccal formulation, a bioavailability study was performed in 12 healthy volunteers who received: a 30 mg oral controlled release tablet (A); a 20 mg aqueous solution retained in the mouth for 10 min (B); and the 60 mg bioadhesive buccal tablet placed between the lower gum and lip for 6 h (C). The mean amount of morphine absorbed from the solution was very low, only 2 mg of the 20 mg dose. After administration of forms A and C, plasma levels exhibit typical sustained release concentration-time curves. The mean amount of drug recovered from the residual bioadhesive buccal tablet after 6 h indicated that approximately 50% of the dose was released from the bioadhesive buccal tablet. The relative bioavailability of the buccal tablet (corrected for residual unabsorbed dose) compared with the controlled-release tablet was 98% based on the morphine AUC values. Good correlations between the AUC and the Cmax of the bioadhesive tablet for the drug and metabolite plotted versus the amount of morphine absorbed were found. PMID- 9737822 TI - Chronopharmacokinetics and calcium in the prevention of gentamicin-induced nephrotoxicity in rabbits. AB - The study used 36 New Zealand white rabbits organized into three groups of 12 animals each. Group I received gentamicin; Group II received joint administration of gentamicin and calcium chloride and Group III received gentamicin, calcium chloride and verapamil. All the drugs were administered over 16 day periods. Groups I and II were divided in two subgroups, one subgroup receiving the treatment in winter and the other in summer. The results obtained for Group I indicate that there is an influence of the seasonal period on the gentamicin elimination and/or distribution. Mean plasma levels of the antibiotic at steady state as well as the amounts of gentamicin accumulated in renal tissue are higher in winter than in summer. On the other hand, when calcium was administrated with the antibiotic, no significant circannual variations were observed in the renal toxicity of gentamicin. Under our study conditions the presence of calcium diminishes gentamicin plasma levels and the amount accumulated in kidney. Calcium, probably, generated a diminution in renal damage and consequently gentamicin renal excretion increases. The differences between Group II and Group III are due to the effect of verapamil. This agent blocks the calcium channels reducing the calcium protective effect on the nephrotoxicity of gentamicin. PMID- 9737823 TI - Blood partition and protein binding of a new reversible proton pump inhibitor, YH1885. PMID- 9737824 TI - Clinical pharmacology of rivastigmine: a new-generation acetylcholinesterase inhibitor for the treatment of Alzheimer's disease. AB - Rivastigmine (ENA 713, or carbamoylatine) is an acetylcholinesterase (AChE) inhibitor with brain-region selectivity and a long duration of action. Both preclinical studies and studies in human volunteers have shown that rivastigmine induces substantially greater inhibition of AChE in the central nervous system (CNS) compartment than in the periphery (40% inhibition of central AChE compared with 10% inhibition of plasma butylcholinesterase in healthy volunteers). Moreover, rivastigmine preferentially inhibits the G1 enzymatic form of AChE, which predominates in the brains of patients with Alzheimer's disease (AD). Evidence from animal studies also suggests that rivastigmine is a more potent inhibitor of AChE in the cortex and hippocampus, the brain regions most affected by AD. Absorption of rivastigmine is rapid and almost complete (>96% of the administered dose). Extensive, saturable first-pass metabolism, however, leads to bioavailability of approximately 35% of the administered dose and nonlinear pharmacokinetics. The principal metabolite of rivastigmine has at least 10-fold lower activity against AChE compared with the parent drug. Rivastigmine is completely metabolized; the major route of elimination of the metabolites is renal. Although patients with AD demonstrate 30% to 50% higher plasma concentrations of rivastigmine and its principal metabolite than do healthy elderly patients, there is no evidence of drug accumulation, which is consistent with rivastigmine's short pharmacokinetic half-life. Distribution of rivastigmine into the CNS is extensive, and inhibition of AChE in the cerebrospinal fluid is detectable 1.2 hours after oral dosing in both healthy volunteers and patients with AD. Peak activity is reached somewhat more slowly in AD patients than in healthy subjects, and the inhibitory effects have a longer duration (6.0 vs 2.4 hours and 12.0 vs 8.5 hours, respectively). Rivastigmine is inactivated during the process of interacting with and inhibiting AChE, and, in contrast to other AChE inhibitors, the hepatic cytochrome P-450 (CYP-450) system is not involved in the metabolism of rivastigmine. This reduces its propensity to interact with drugs metabolized by specific CYP-450 isoenzymes. Consistent with rivastigmine's pharmacokinetic and pharmacodynamic profiles, Phase II and III trials have demonstrated that the drug is a well-tolerated and effective treatment for AD. PMID- 9737825 TI - The interpretation of preclinical data in predicting bisphosphonate response in the treatment of osteoporosis. AB - Individual bisphosphonates for the treatment of osteoporosis are often compared on the basis of preclinical variables such as potency, selectivity, therapeutic index, and effects on fracture healing. The present review examines the methodology and results of preclinical studies providing these types of data for the bisphosphonate etidronate and compares them with the available preclinical data for other bisphosphonates. Analysis of the preclinical data in relation to the results of long-term clinical trials reveals that widely reported differences in preclinical variables for the different bisphosphonates are not significant once dosages and treatment regimens have been optimized for therapeutic use in the clinic. PMID- 9737826 TI - Advances in the treatment of herpesvirus infection: the role of famciclovir. AB - Shingles (herpes zoster) is the result of reactivation of varicella-zoster virus after years of latency. The acute phase is self-limiting but is often associated with moderate-to-severe pain; postherpetic neuralgia is the most frequent and debilitating complication of shingles, occurring in 3.4 per 1000 individuals per year. In the case of genital herpes, herpes simplex virus can reactivate to cause recurrent episodes as often as several times a year, sometimes for the remainder of a person's life. Antiviral agents such as famciclovir, valacyclovir, and acyclovir can be used to shorten the course and decrease the severity of these diseases and may suppress the virus itself, thereby preventing future outbreaks of genital herpes. This article presents a brief synopsis of the etiology of herpes zoster and genital herpes and reviews 12 key studies that demonstrate the efficacy of famciclovir in the management of these two conditions. PMID- 9737827 TI - Continuation of initial antihypertensive medication after 1 year of therapy. AB - This paper describes use of the prescription records of a large pharmaceutical benefits management organization to retrospectively analyze the refill behavior of patients who have recently started antihypertensive therapy in the outpatient setting. Using logistic regression analysis, the author identified class of antihypertensive medication, patient age, and dosing frequency as clinically important independent covariates that are predictive of persistence (defined as continuing therapy with the original antihypertensive drug as originally prescribed) at 12 months. At 12 months' follow-up, the percentage of patients continuing initial angiotensin II (A-II) antagonist therapy was substantially higher than the percentage continuing therapy with angiotensin-converting enzyme inhibitors, calcium antagonists, beta-blockers, or thiazide diuretics (64% vs 58%, 50%, 43%, and 38%, respectively). Additional studies are needed to explain why more patients continued with the same A-II antagonist therapy at 12 months compared with the other classes of antihypertensive drugs; whether these findings are explained by drug tolerability, financial incentives, newness of the product, selection bias, or other factors; whether these differences will be maintained in the following years; and whether the differences are associated with better health outcomes. PMID- 9737828 TI - Determination of the dose proportionality of single intravenous doses (5, 10, and 15 mg) of lubeluzole in healthy volunteers. AB - The dose proportionality of lubeluzole, a drug in clinical development for the treatment of acute ischemic stroke, was evaluated in a Phase I, single-center, open-label, randomized-dosing-sequence, three-way crossover clinical trial in 12 healthy adults. An equal number of male and female volunteers were enrolled in the trial, with a mean weight (+/- SD) of 73.2 +/- 11.9 kg, mean height (+/- SD) of 66.8 +/- 4.6 inches, and a mean age of 36.1 +/- 5.2 years. Subjects received intravenous infusions of 5 (A), 10 (B), and 15 mg (C) of lubeluzole over 1 hour on three separate occasions, with a minimum washout period of 2 weeks. The treatment sequences were A-B-C; A-C-B; B-A-C; B-C-A; C-A-B; and C-B-A. One male and one female were assigned to each sequence. After the 5-, 10-, and 15-mg doses, maximum concentration (Cmax) was 58.1, 113, and 138 microg/L, respectively, and the area under the curve from 0 to (AUC(0-infinity)) was 771, 1384, and 2025 microg x h/L. There were no statistically significant differences among the groups in mean terminal half-life, steady-state volume of distribution, total plasma clearance, or dose-normalized AUC(0-infinity). Dose-normalized values of Cmax differed significantly between the groups. No serious adverse events were reported, and no changes were observed in cardiac function, as judged by the QT(c) interval. The pharmacokinetics of lubeluzole appear to be linear at intravenous infusion doses of 5, 10, and 15 mg, and these doses are well tolerated by healthy adults. PMID- 9737829 TI - A retrospective chart review of uncontrolled use of metformin as an add-on therapy in type 2 diabetes. AB - The hyperglycemia, hyperinsulinemia, insulin resistance, and obesity syndrome associated with type 2 diabetes can have debilitating consequences. The biguanide metformin has a mechanism of action that is complementary to those of insulin and the sulfonylureas, suggesting that combination therapy that includes metformin may result in improved glycemic control. The purpose of this retrospective chart review was to determine the effects of adding metformin in an uncontrolled fashion to existing therapy in obese patients with type 2 diabetes who had suboptimal glycemic control and insulin resistance. For the review, the records of 124 patients were divided into two groups: group 1 included 71 patients who were taking insulin with or without a sulfonylurea, and group 2 consisted of 53 patients who were taking a sulfonylurea alone. Metformin was added to patients' existing therapy in conjunction with downward titration of the sulfonylurea and insulin doses. A retrospective chart review was conducted at the end of 6 months for group 1 and at the end of 12 months for group 2 to determine the change from baseline in measures of diabetes control (ie, insulin and sulfonylurea dose, glycated hemoglobin [Hb A1c] value, body mass index [BMI], and lipid profiles). In group 1, the mean insulin dose decreased from 46.4 U/d at baseline to 6.1 U/d at the end of follow-up. Eighty-three percent of the patients were able to discontinue insulin therapy completely. Similarly, group 2 had statistically significant reductions in mean sulfonylurea dose. Both groups also achieved statistically significant reductions in Hb A1c, BMI, and total cholesterol level. The addition of metformin to treatment with insulin or sulfonylureas, either alone or in combination, significantly improved glycemic control and cholesterol levels and promoted weight loss in obese type 2 diabetic patients with insulin resistance. Less than 5% of patients reported mild, transient gastrointestinal side effects, none of which required cessation of metformin therapy. Five patients discontinued metformin due to lack of efficacy. PMID- 9737830 TI - Weight-based heparin dosing: clinical response and resource utilization. AB - The objective of this study was to assess a weight-based heparin (WBH) nomogram (80-U/kg bolus, 18-U/kg-per-hour initial infusion) and determine its clinical performance and impact on resource utilization. All patients treated with heparin for venous thromboembolism or unstable angina during a 15-week study period were included in this retrospective, chart-review study. Three groups were identified: patients treated with WBH, patients whose regimen deviated from the weight-based nomogram (DEV), and matched historical controls (HCs). In patients receiving heparin for more than 24 hours, those treated with WBH achieved threshold activated partial thromboplastin time (aPTT) levels significantly faster than did HC or DEV patients. However, 42% of WBH-treated patients were found to have initial supratherapeutic responses. Logistic regression analysis identified age > or =67 years, prior warfarin therapy within 7 days of heparin, and high initial infusion rate as predictive of a supratherapeutic aPTT response; smoking was predictive of a subtherapeutic response. Bleeding events were not significantly different between groups. An infusion rate of 15 U/kg per hour was found to closely approximate our population's actual heparin infusion requirement. Resource utilization was significantly different between the WBH and HC groups in terms of nursing interventions at 48 to 72 hours. We concluded that WBH rapidly drives patients' aPTT response above the therapeutic threshold for heparin; however, prudent adjustment of the initial infusion rate is necessary to avoid a supratherapeutic aPTT response. Our data support a nomogram with an initial infusion rate of 15 U/kg per hour. PMID- 9737831 TI - Efficacy and safety of azelaic acid and glycolic acid combination therapy compared with tretinoin therapy for acne. AB - We conducted a 12-week, multicenter, randomized, double-masked, parallel-group study of the efficacy, safety, and tolerability of azelaic acid 20% cream and glycolic acid lotion compared with tretinoin 0.025% cream and a vehicle lotion to treat mild-to-moderate facial acne vulgaris. Patients treated with azelaic/glycolic acid experienced a significantly greater reduction in the number of papules, as well as a greater reduction in the number of inflammatory lesions, than those treated with tretinoin. Overall global improvement was approximately 25% in both groups. In the physician evaluations, treatment with azelaic/glycolic acid was found to cause significantly less dryness, scaling, and erythema than tretinoin. Patients also reported significantly less dryness, redness, and peeling with azelaic/glycolic acid. Significantly more patients in the azelaic/glycolic acid group than the tretinoin group reported that they felt attractive. The combination of azelaic acid and glycolic acid is a useful alternative to tretinoin, being at least as efficacious as the latter, while offering a superior tolerability and patient approval profile. PMID- 9737832 TI - Effect of escalating doses of recombinant human granulocyte colony-stimulating factor (filgrastim) on circulating neutrophils in healthy subjects. AB - The safety profile, tolerability, pharmacodynamics, and pharmacokinetics of four doses of recombinant human granulocyte colony-stimulating factor (filgrastim) were assessed in healthy volunteers in a double-masked, placebo-controlled, parallel-group trial. Healthy subjects received subcutaneous injections of filgrastim 75 microg (n = 8), 150 microg (n = 4), 300 microg (n = 4), 600 microg (n = 8), or placebo (n = 6) daily for 10 consecutive days. Blood samples were drawn daily immediately before the injection and on days 1 and 10 serially throughout the day. Increased absolute neutrophil counts (ANCs) were seen within 90 minutes of drug administration in subjects in all dose groups, peaking approximately 12 hours after administration. This increase was dose related in subjects in the three lower dose groups. The time to peak ANC on day 10 was approximately 9 hours, with a daily ANC profile in all four dose groups that was similar to the profile on day 1. In all dose groups, ANCs were near baseline within 48 hours of discontinuation of filgrastim. Mild, reversible thrombocytopenia was reported in 4 of 10 subjects in the highest dose group. Two subjects in the filgrastim 600-microg group were withdrawn for adverse events. Filgrastim had a good safety profile and caused dose-related increases in ANC when administered to healthy volunteers for up to 10 days. PMID- 9737833 TI - Comparison of dextranomer paste and saline dressings for management of decubital ulcers. AB - In this open-label, parallel-group study, 23 male spinal cord injury patients aged 23 to 73 years (median 54 years) with a total of 30 exudative decubital ulcers were randomly allocated to receive treatment with dextranomer paste (15 ulcers) or conventional saline dressings (15 ulcers). Treatment was applied at least once every 12 hours and continued for a maximum of 15 days, until the ulcer was clean and covered with new granulation tissue and was suitable for skin grafting. The same physician assessed ulcer status (extent of drainage, granulation, erythema, and edema and the presence or absence of necrosis and epithelialization) each time the study nurse changed the dressings. Treatment with dextranomer paste resulted in significantly greater improvement in ulcer drainage compared with saline; 73% versus 13% of the ulcers, respectively, showed > or =25% improvement in drainage from baseline to the end of the study. Both treatments were well tolerated, with no evidence of any local irritation. Dextranomer paste was more effective than saline dressings in the management of decubital ulcers in spinal cord injury patients. PMID- 9737834 TI - Clinical trial of ototopical ofloxacin for treatment of chronic suppurative otitis media. AB - A multicenter, open-label prospective trial was performed to determine the clinical and microbiologic efficacy of ofloxacin (OFLX) otic solution in the treatment of subjects > or =12 years with chronic suppurative otitis media (CSOM) and a chronically perforated tympanic membrane in the infected ear(s). A total of 207 patients at 27 centers in the United States and Central America received OFLX 0.5 mL instilled ototopically twice daily for 14 consecutive days. The primary clinical end point was cure (dry ear) or failure (not dry ear). The primary microbiologic end point was eradication of baseline pathogens. Because there was no comparator and there were few data in the literature regarding clinical efficacy in patients treated with other regimens, the efficacy of OFLX was compared with data recorded in the clinical records of historical-practice control (HPC) or current-practice control (CPC) subjects. The incidence of clinical cure in clinically evaluable OFLX-treated patients (91%; 148 of 162 subjects) was significantly higher than in HPC subjects (67%; 124 of 185 subjects) or CPC subjects (70%; 38 of 54 subjects). OFLX eradicated all baseline pathogens isolated in microbiologically evaluable subjects. These pathogens were predominantly Staphylococcus aureus, Pseudomonas aeruginosa, and Proteus mirabilis. The most common treatment-related adverse event, bitter taste, occurred in 17% (35 of 207) of OFLX-treated subjects. Thus OFLX 0.5 mL administered twice daily for 14 days was effective in resolving the signs and symptoms of CSOM in subjects > or =12 years, was significantly more effective than therapies used to treat HPC or CPC subjects, and was well tolerated. PMID- 9737835 TI - Predictors of medication adherence in the elderly. AB - As average life expectancy increases, so do the incidence of chronic diseases and the number of persons receiving long-term drug therapy. Thus elderly patients' noncompliance with medication regimens has the potential for sweeping medical and economic consequences and is likely to become increasingly important in the design of disease-management programs for this population. The author conducted a MEDLINE search of the English-language literature for the years 1962 to 1997 to identify articles concerning predictors of medication compliance in the elderly. A descriptive analysis of this literature indicated that there remains some uncertainty about the reasons for noncompliant medication-taking in the elderly. Clear associations have been established between elderly patients' medication adherence and race, drug and dosage form, number of medications, cost of medications, insurance coverage, and physician-patient communication. However, the findings are inconsistent with regard to the effects of patients' age, sex, socioeconomic status, living arrangement, comorbidities, number of physician visits, and knowledge, attitudes, and beliefs about health. Until the results of further comprehensive studies are available, the current knowledge should be considered when designing and implementing disease-management programs for the elderly. PMID- 9737836 TI - A review of cost-of-illness studies on obesity. AB - This paper reviews the published cost-of-illness studies on obesity. The medical literature has demonstrated that obesity is an independent risk factor for a number of medical conditions, including diabetes mellitus, hypertension, coronary heart disease, elevated cholesterol levels, depression, musculoskeletal disorders, gallbladder disease, and several cancers. Since these conditions can be costly to treat, obesity clearly has a substantial economic impact. Epidemiologic estimates of the aggregate economic costs associated with specific obesity-related diseases in the United States indicate that the annual burden to society totals in the billions of dollars, representing 5.5% to 7.8% of total health-care expenditures. Although estimates of the costs attributable to obesity differ across studies, the one common finding is that these costs are substantial from a health-policy perspective. The objective of this paper is to identify and review the obesity cost-of-illness literature, address study limitations, and identify key areas for future economic research. This review indicates that the economic burden of obesity has been estimated using a prevalence-based cost-of illness framework. Areas for future research include estimating the economic burden of obesity using an incidence-based cost-of-illness framework and modeling the association between health-care expenditure and level of obesity using individual-level data, such as medical and pharmacy claims data. PMID- 9737837 TI - Economic consequences of selective serotonin reuptake inhibitor use with drugs also metabolized by the cytochrome P-450 system. AB - Administration of selective serotonin reuptake inhibitors (SSRIs) may increase plasma concentrations of concomitant medications that are also metabolized by the cytochrome P-450 system (CYP-450), in particular by the 2D6 and 3A4 isoenzymes. This may lead to side effects or other clinical events that might be expected to incur higher health-care expenditures. The purpose of this study was to assess whether there was a difference in expenditures during the first 90 days of SSRI therapy with paroxetine or sertraline versus fluoxetine in patients who were also receiving a stable dosage of a nonpsychiatric drug also metabolized by the CYP 450 2D6 or 3A4 isoenzyme systems. A sample of 2445 patients who initiated therapy with an SSRI while receiving a stable dosage of a nonpsychiatric drug was obtained from a private insurance claims database. Multivariate regression techniques were used to estimate total health-care expenditures in the first 90 days after receiving a prescription for an SSRI. After adjusting for nonrandom SSRI prescription patterns and controlling for observable and unobservable characteristics that might correlate with SSRI selection, total health-care expenditures were 95% higher for patients initiating SSRI therapy with sertraline or paroxetine compared with fluoxetine. Results suggest that there are cost differences between SSRIs during concomitant therapy with drugs also metabolized by the CYP-450 system. To determine whether there are additional differences in expenditures across SSRIs, future research should focus on (1) simultaneous initiation of SSRI therapy and a nonpsychiatric drug also metabolized by the CYP 450 enzyme system, and (2) addition of nonpsychiatric drug therapy to stable SSRI therapy. Relationships between additional expenditures, drug interactions, and clinical outcomes should also be assessed directly using medical records and patient interview data that are not available in claims-based files. PMID- 9737838 TI - Selective serotonin reuptake inhibitor utilization patterns: consistency across research designs. AB - We examined the impact of commonly applied selection criteria on the ability of patients who are initiating antidepressant therapy to reach a stable pattern, which was defined as receipt of only the initial agent at the initial dose for 90 or more consecutive days. Patients in a large US prescription database who initiated fluoxetine, paroxetine, or sertraline therapy between February and April of 1995 were categorized as with (typical design) and without (relaxed design) commonly applied selection criteria. The percentage of patients achieving a stable pattern was then determined. We found that this percentage was significantly higher with the relaxed design (typical, 28.8%; relaxed, 32.4%) and for patients initiating fluoxetine therapy (>5.5% higher than for those initiating paroxetine or sertraline therapy). The results for fluoxetine were consistent across designs, whereas comparisons between paroxetine and sertraline yielded mixed results. Therefore, the relative relationship of the stable pattern is robust across designs for fluoxetine but not for paroxetine and sertraline. Further, application of commonly applied selection criteria may make a sample less representative and reduce the measured rates of stable antidepressant use, potentially leading to underestimation of the benefits of pharmacotherapy. PMID- 9737839 TI - Are state immunization programs effective? Implications for the children's immunization initiative. AB - Several states operate universal vaccine purchase (UVP) programs aimed at ensuring adequate immunization of children. Some of these programs have been in operation for many decades. Recently, there has been a great deal of interest in implementing a federal UVP program. It is not clear, however, that such programs can significantly increase immunization levels; many nonfinancial barriers to full immunization exist and would have to be addressed. This paper uses cross sectional data at the state level to estimate the effect of state UVP programs on the immunization levels of preschool children. The results indicate that states with UVP programs do not have significantly higher immunization rates than do other states. Therefore, it is not likely that a federal UVP program would significantly affect immunization rates. PMID- 9737840 TI - The cost of treating community-acquired pneumonia. AB - Community-acquired pneumonia (CAP) is responsible for an average of 4.5 million visits annually to physicians' offices, emergency departments, and outpatient clinics. However, there have been few studies using national data on the costs of treating CAP. Without such data, it is difficult to assess whether new therapies and treatment strategies are needed to improve patient outcomes. We conducted a retrospective analysis based on national incidence data and paid claims data for patients treated for CAP to assess the frequency of services rendered and costs to the health-care system. Records were selected for the study based on a primary diagnosis of CAP according to the International Classification of Diseases, 9th Revision. Incidence data were derived from the National Health and Nutrition Examination Survey III. Medicare was the primary source of data for patients aged > or =65 years. Data from the National Healthcare Cost and Utilization Project, the National Ambulatory Medical Care Survey, and the National Hospital Ambulatory Medical Care Survey were used to determine the cost of treating patients aged <65 years. We arrived at a total cost of $4.8 billion for treating patients aged > or =65 years and $3.6 billion for treating patients aged <65 years. These calculations were based on the following: 1.1 million hospital discharges resulting in inpatient costs of $4.4 billion (52.4% of the $8.4 billion) for the 0.6 million patients aged > or =65 years and $3.1 billion (36.9% of the $8.4 billion) for the 0.5 million patients aged <65 years. The average hospital length of stay was 7.8 days with an average cost of $7166 for patients aged > or =65 years and 5.8 days with an average cost of $6042 for younger patients. Room and board represented the largest percentage of the average hospital bill for patients with CAP. Inpatient physician service costs were $305 million and $192 million for the > or =65 and <65 groups, respectively. Based on 1.1 million outpatient office visits for those aged > or =65 years and 3.3 million visits for those aged <65, total outpatient costs were $119 million and $266 million, respectively. Given the overwhelming cost burden for CAP in the hospital setting, any new therapy that allows patients to be treated in the outpatient setting could result in significant savings, especially for patients aged > or =65 years. PMID- 9737841 TI - An economic evaluation of donepezil in the treatment of Alzheimer's disease. AB - Using data from a longitudinal survey of caregivers of Alzheimer's disease patients, we calculated the average per-patient direct medical costs over a 6 month period for a matched sample of patients (N = 376). A group of patients receiving donepezil for 6 months was compared with a group not receiving this form of drug therapy. The groups were matched by disease severity, age, sex, and comorbidity. The average age in the two groups was 74 years, with 50% female and 90% white. Patients in both groups had a mean of 1.6 comorbid conditions. No patients in either group were institutionalized at the beginning of the 6-month period, and all patients were taking at least one prescription drug, including donepezil. Mean 6-month direct medical expenses for a patient receiving donepezil were $3443, including the cost of the drug, whereas the per-patient mean expenses for the comparison group were $3476. Although the patients receiving donepezil had greater expenditures for prescription drugs, these costs were offset by a slower rate of institutionalization. At the end of the 6-month period, 5% of donepezil patients were institutionalized, compared with 10% of the nondonepezil patients. The cost of receiving donepezil treatment for 6 months did not result in a significantly higher per-patient mean direct cost. PMID- 9737842 TI - A pharmacoeconomic analysis of topical therapies for patients with mild-to moderate stable plaque psoriasis: a US study. AB - Psoriasis is a persistent skin disorder characterized by abnormal keratinocyte differentiation, keratinocyte hyperproliferation, and increased expression of inflammatory markers at the cellular level, leading to erythema, induration, and scaling of the skin. Depending on the severity of the disease, annual outpatient costs range from $1400 to $6600 per patient, totaling $3.2 billion each year in the United States. Because the disease is persistent and progressive, patients receiving a diagnosis of psoriasis early in life can expect to require lifelong care, which translates into lifelong expense. Treatments include topical formulations, systemic therapies, phototherapies, and combination therapies. Of these, topical agents are the first-line treatments, including fluocinonide and other steroids, calcipotriene, and tazarotene, a once-daily retinoid. To establish the relative cost-effectiveness of these drugs (fluocinonide, calcipotriene, and tazarotene), we conducted a pharmacoeconomic study from the perspective of a third-party payer, using a decision-analytic model validated by clinical experts. Data were drawn from a meta-analysis of the contemporary medical literature. Clinical success, clearing, and relapse rates determined the probabilities for therapeutic outcomes and the number of anticipated disease-free days for each study comparator. Costs for physician visits, drug acquisition, laboratory testing, and adverse-events management were added to each branch of the decision tree and multiplied by the appropriate probabilities to establish the expected cost of treatment, stratified by the primary treatment choice. Cost effectiveness was expressed as the total expected cost of achieving a disease free day. Tazarotene 0.1% was 16.74% more cost-effective than tazarotene 0.05%, 85.46% more cost-effective than fluocinonide, and 143.75% more cost-effective than calcipotriene. The expected cost of achieving a disease-free day was $49.46 for tazarotene 0.1%, $57.74 for tazarotene 0.05%, $91.73 for fluocinonide, and $120.56 for calcipotriene. Treatment with tazarotene offers an opportunity to reduce the cost of care for patients with mild-to-moderate psoriasis and enhance patient satisfaction by gaining more disease-free days. PMID- 9737843 TI - Trends in the prescribing of antidepressant pharmacotherapy: office-based visits, 1990-1995. AB - Data from the National Ambulatory Medical Care Survey for the period 1990 through 1995 were used to discern the population-adjusted rate of office-based physician patient encounters at which the prescribing or continuation of antidepressant pharmacotherapy (tricyclic antidepressants [TCAs], selective serotonin reuptake inhibitors [SSRIs], or others), a diagnosis of depression (International Classification of Diseases, 9th Revision, Clinical Modification codes 296.2 through 296.36, 300.4, or 311), or both were documented. National estimates of the number of office-based visits resulting in a prescription for or continuation of antidepressant pharmacotherapy for any purpose escalated from 16,534,268 in 1990 to 28,664,796 in 1995, a 73.4% increase. Although the number of office-based visits at which a diagnosis of depression was documented increased 23.2% during this period, the proportion of patients with a diagnosis of depression who were prescribed or continued antidepressant pharmacotherapy increased only 14.9%, from 52.1% in 1990 to 67.0% in 1995. Among patients with a diagnosis of depression, use of a TCA declined from 42.1% in 1990 to 24.9% in 1995. In contrast, use of an SSRI for the treatment of depression increased from 37.1% in 1990 to 64.6% in 1995. The rate of office-based visits at which the use of antidepressant pharmacotherapy for any purpose was documented increased from 6.7 per 100 US population in 1990 to 10.9 in 1995, a 62.7% increase; documentation of a diagnosis of depression increased from 6.1 per 100 US population in 1990 to 7.1 in 1995, a 16.4% increase; and the recording of a diagnosis of depression in concert with the prescribing or continuation of antidepressant pharmacotherapy increased from 3.2 per 100 US population in 1990 to 4.8 in 1995, a 50.0% increase. Further research is required to elucidate the effect of observed trends on clinical and financial outcomes. PMID- 9737844 TI - Identification of protein folding patterns using site-directed spin labeling. Structural characterization of a beta-sheet and putative substrate binding regions in the conserved domain of alpha A-crystallin. AB - The folding pattern of the segment of alphaA-crystallin encoded by exon 2 and containing putative substrate binding sites was explored using site-directed spin labeling (SDSL). For this purpose, a nitroxide scan was carried out between residues 60 and 108. At each site, structural constraints describing the local environment and topography were obtained from analysis of the nitroxide mobility and its solvent accessibility. Periodic patterns in the sequence-specific variation of these parameters were used to assign the secondary structure along the sequence. Geometric constraints describing the packing of secondary structure were deduced from patterns of proximities in 20 nitroxide pairs, specifically designed to differentiate between supersecondary structural motifs. Our data, in conjunction with those of Berengian et al. [Berengian, A. R., Bova, M. P., and Mchaourab, H. S. (1997) Biochemistry 36, 9951-9957], reveal that the fold of the segment between residues 84 and 120 consists of an antiparallel beta-sheet of three strands arranged in consecutive beta-hairpins. The boundaries of the sheet are defined at one end by a surface of isologous association and on the other end by an unstructured, charged interdomain segment. One of the putative substrate binding segments overlaps a buried loop, suggesting that the structural origin of the thermal activation of binding is the transient exposure of this site. This paper describes and implements a general strategy for experimental fold recognition using SDSL. The results of its application to alphaA-crystallin provide the first experimental insight into the folding pattern of the subunit and establish the structural context necessary to understand molecular recognition and substrate binding. PMID- 9737845 TI - Solution structure and backbone dynamics of the photoactive yellow protein. AB - The solution structure of photoactive yellow protein (PYP), a photosensory protein from Ectothiorhodospira halophila, has been determined by multidimensional NMR spectroscopy. The structure consists of an open, twisted, 6 stranded, antiparallel beta-sheet, which is flanked by four alpha-helices on both sides. The final set of 26 selected structures is well-defined for the regions spanning residues Phe6-Ala16, Asp24-Ala112, and Tyr118-Val125 and displays a root mean-square deviation, versus the average, of 0.45 A for the backbone and 0.88 A for all heavy atoms. Comparison of the solution structure with an earlier published 1.4 A crystal structure (Borgstahl, G. E. O., Williams, D. R., and Getzoff, E. D. (1995) Biochemistry 34, 6278-6287) reveals a similarity with a root-mean-square deviation of 1.77 A for the backbone for the well-defined regions. The most distinct difference in the backbone with the crystal structure is found near the N-terminus, for residues Asp19-Leu23, which corresponds to an alpha-helix in the crystal structure and to one of the poorest defined regions in the solution structure. To characterize the dynamic behavior of PYP in solution, we undertook a 15N relaxation study and measurements of hydrogen/deuterium exchange. Determination of order parameters through the model-free Lipari-Szabo approach enabled the identification of several regions of enhanced dynamics. The comparison of atomic displacements in the backbone traces of the ensemble structures, with mobility measurements from NMR, show that the poorly defined regions feature fast internal motions in the nanosecond to picosecond time scale. PMID- 9737846 TI - Solution structure of methionine-oxidized amyloid beta-peptide (1-40). Does oxidation affect conformational switching? AB - The solution structure of Abeta(1-40)Met(O), the methionine-oxidized form of amyloid beta-peptide Abeta(1-40), has been investigated by CD and NMR spectroscopy. Oxidation of Met35 may have implications in the aetiology of Alzheimer's disease. Circular dichroism experiments showed that whereas Abeta(1 40) and Abeta(1-40)Met(O) both adopt essentially random coil structures in water (pH 4) at micromolar concentrations, the former aggregates within several days while the latter is stable for at least 7 days under these conditions. This remarkable difference led us to determine the solution structure of Abeta(1 40)Met(O) using 1H NMR spectroscopy. In a water-SDS micelle medium needed to solubilize both peptides at the millimolar concentrations required to measure NMR spectra, chemical shift and NOE data for Abeta(1-40)Met(O) strongly suggest the presence of a helical region between residues 16 and 24. This is supported by slow H-D exchange of amide protons in this region and by structure calculations using simulated annealing with the program XPLOR. The remainder of the structure is relatively disordered. Our previously reported NMR data for Abeta(1-40) in the same solvent shows that helices are present over residues 15-24 (helix 1) and 28 36 (helix 2). Oxidation of Met35 thus causes a local and selective disruption of helix 2. In addition to this helix-coil rearrangement in aqueous micelles, the CD data show that oxidation inhibits a coil-to-beta-sheet transition in water. These significant structural rearrangements in the C-terminal region of Abeta may be important clues to the chemistry and biology of Abeta(1-40) and Abeta(1-42). PMID- 9737847 TI - Tertiary structure of the major house dust mite allergen Der p 2: sequential and structural homologies. AB - Sensitization to indoor allergens, especially those of the house dust mite, is strongly correlated with the development of asthma. We report the tertiary structure of the major house dust mite allergen, Der p 2, determined by NMR methods. The structure of Der p 2 is a beta-barrel and is composed of two three stranded antiparallel beta-pleated sheets. This arrangement of beta-strands is similar to the immunoglobulin fold with respect to the orientation of the two sheets and the interactions of the strands. However, the three-dimensional structure of Der p 2 aligns equivalently with a number of proteins from different families within the immunoglobulin superfamily. The structural homology with the highest significance score from analysis by DALI is to Der f 2. Although Der p 2 and Der f 2 are 87% identical in amino acid sequence, they align in three dimensions rather poorly (4.85 A RMSD; Z-score, 8.58). This unexpected finding is likely due to the different solution conditions used during structure determination by NMR for both proteins. While the structural comparisons did not elucidate a clear homologue for the function of Der p 2 in mites, we report that Der p 2 is sequentially homologous to esr16. This is a protein from moths that is expressed coincident with molting. Thus, this homology has important ramifications for the study of mite allergy. The structure of Der p 2 provides a useful tool in the design of recombinant immunotherapeutics for the group 2 allergens. PMID- 9737848 TI - A pseudosquare knot structure of DNA in solution. AB - We report a high-resolution NMR structure of a homodimer formed by a synthetic 25 residue DNA oligonucleotide GCTCCCATGGTTTTTGTGCACGAGC. This structure presents a novel structural motif for single-stranded nucleic acids, called a pseudosquare knot (PSQ). The oligonucleotide was originally designed to mimic a slipped-loop structure (SLS), another "unusual" DNA structure postulated as an alternative conformation for short direct repeats in double-stranded DNA. The design of the sequence is compatible with both SLS and PSQ structures, both of which possess identical sets of base-paired and unpaired nucleotides but different tertiary folds. We used deuteration of the H8 positions of purines to ascertain that the PSQ is actually formed under the conditions used. The PSQ structure was solved based on homonuclear proton nuclear Overhauser effect data using complete relaxation matrix methods. The structure essentially consists of two side-by-side helices connected by single-stranded loops. Each of the helices is well-defined; however, the relative orientation of the two remains undetermined by the NMR data. The sequences compatible with the PSQ formation are frequent in single stranded genomes; this structure may play a role as a dimerization motif. PMID- 9737849 TI - NMR solution structure of type II human cellular retinoic acid binding protein: implications for ligand binding. AB - The structure of human apo-cellular retinoic acid binding protein II (apo CRABPII) in solution at pH 7.3 has been determined by NMR spectroscopy. The sequential assignments of the 1H, 13C, and 15N resonances of apo-CRABPII were established by multinuclear, multidimensional NMR spectroscopy. The solution structure of apo-CRABPII was derived from 2382 experimental NMR restraints using a hybrid distance geometry-simulated annealing protocol. The root-mean-square deviation of the ensemble of 25 refined conformers that represent the structure from the mean coordinate set derived from them was 0.54 +/- 0.18 and 0.92 +/- 0.20 A for the backbone atoms and all heavy atoms, respectively, of all residues except Ala32-Pro39 and Thr57-Glu62, which are in disordered regions. The solution structure of apo-CRABPII is similar to the crystal structure of holo-CRABPII [Kleywegt, G. J., Bergfors, T., Senn, H., Le Motte, P., Gsell, B., Shudo, K., and Jones, T. A. (1994) Structure 2, 1241-1258] except the ligand entrance, which is sufficiently enlarged in the apoprotein to be readily accessible to retinoic acid. The enlargement of the ligand entrance of apo-CRABPII relative to that of holo-CRABPII is due mainly to a concerted conformational change in three structural elements, namely, the second helix, the betaC-betaD loop, and the betaE-betaF loop. Furthermore, the ligand-binding pocket of apo-CRABPII showed evidence of dynamic disorder; among the 21 residues that constitute this pocket, 16 residues had weak or no detectable cross-peaks in the two-dimensional 1H-15N HSQC spectrum recorded under conditions of minimal water saturation or dephasing. Apo-CRABPII is largely monomeric in solution, with no evidence for the dimeric structure shown in the crystal structure of apo-CRABPI which was suggested to be a prerequisite for ligand entry [Thompson, J. R., Bratt, J. M., and Banaszak, L. J. (1995) J. Mol. Biol. 252, 433-446]. Thus, the widening of the ligand entrance required for entry of retinoic acid appears to be a property of monomeric apo CRABPII. PMID- 9737850 TI - Binding domain of human parathyroid hormone receptor: from conformation to function. AB - A 31 amino acid fragment of the extracellular N-terminus of the human G-protein coupled receptor for parathyroid hormone (PTH1R) has been structurally characterized by NMR and molecular dynamics simulations. The fragment PTH1R[168 198] includes residues 173-189, shown by photoaffinity cross-linking to be a contact domain with position 13 of parathyroid hormone (PTH). The structure of PTH1R[168-198], determined in a micellar solution of dodecylphosphocholine to mimic the membrane environment, consists of three alpha-helices, separated by a well-defined turn and a flexible region. The topological orientation of PTH1R[168 198] was determined from nitroxide-radical induced relaxation of NMR signals utilizing 5- and 16-doxylstearic acid. The C-terminal helix (residues 190-196), consisting of seven amino acids of the first transmembrane domain, is very hydrophobic and embedded in the lipid core. This helix is preceded by a well defined turn, forming an approximate 90 degrees bend, placing the other helices (residues 169-176 and 180-189), both of which are amphipathic, on the surface of the micelle. In this orientation, many hydrophilic residues of the receptor, including Glu177, Arg179, Arg181, Glu182, Asp185, and Arg186, are projecting toward the solvent available to form complementary Coulombic interactions with the polar residues of the principal binding domain of the ligand (e.g., Arg25, Lys26, Lys27, Asp30, and His32). Given that the binding domain of PTH adopts an amphipathic alpha-helix which lies on the membrane, we visualize ligand binding as a two stage process involving a nonspecific hydrophobic interaction of amphipathic helices with the membrane, followed by two-dimensional diffusion leading to highly specific, ligand-receptor interaction. PMID- 9737851 TI - Isolation and characterizations of quinone analogue-resistant mutants of bo-type ubiquinol oxidase from Escherichia coli. AB - Cytochrome bo is a member of the heme-copper terminal oxidase superfamily and serves as a four-subunit ubiquinol oxidase in the aerobic respiratory chain of Escherichia coli. To probe the location and structural properties of the ubiquinol oxidation site, we isolated and characterized five or 10 spontaneous mutants resistant to either 2,6-dimethyl-1,4-benzoquinone, 2,6-dichloro-4 nitrophenol, or 2,6-dichloro-4-dicyanovinylphenol, the potent competitive inhibitors for the oxidation of ubiquinol-1 [Sato-Watanabe, M., Mogi, T., Miyoshi, H., Iwamura, H., Matsushita, K., Adachi, O., and Anraku, Y. (1994) J. Biol. Chem. 269, 28899-28907]. Analyses of the growth yields and the ubiquinol-1 oxidase activities of the mutant membranes showed that the mutations increased the degree of the resistance to the selecting compounds. Notably, several mutants showed the cross-resistance. These data indicate that the binding sites for substrate and the competitive inhibitors are partially overlapped in the ubiquinol oxidation site. All the mutations were linked to the expression vector, and 23 mutations examined were all present in the C-terminal hydrophilic domain (Pro96-His315) of subunit II. Sequencing analysis revealed that seven mutations examined are localized near both ends of the cupredoxin fold. Met248Ile, Ser258Asn, Phe281Ser, and His284Pro are present in a quinol oxidase-specific (Qox) domain and proximal to low-spin heme b in subunit I and the lost CuA site in subunit II, whereas Ile129Thr, Asn198Thr, and Gln233His are rather scattered in a three-dimensional structure and closer to transmembrane helices of subunit II. Our data suggest that the Qox domain and the CuA end of the cupredoxin fold provide the quinol oxidation site and are involved in electron transfer to the metal centers in subunit I. PMID- 9737852 TI - A loop deletion in the plant acetohydroxy acid isomeroreductase homodimer generates an active monomer with reduced stability and altered magnesium affinity. AB - Plant acetohydroxy acid isomeroreductase is a stable homodimer which catalyzes in the presence of magnesium an alkyl migration followed by a NADPH-dependent reduction. Since the enzyme exhibits no kinetic cooperativity either for its cofactor (NADPH and magnesium) or for its substrates, the reason for dimerization of this enzyme was not obvious. Recently, crystallographic studies [Biou, V., et al. (1997) EMBO J. 16, 3405-3415] revealed that the loop of residues 422-431 plays a major part in the dimer interface. To understand the role of the quaternary structure of the enzyme, we have deleted residues 423-430 and substituted Phe 431 for serine. This mutant was further overproduced in Escherichia coli, purified to homogeneity, and characterized. Gel filtration and thermodynamic experiments disclosed that this mutant behaves as an active monomer with reduced thermal stability. Furthermore, kinetic and fluorescence experiments showed that the behavior of the monomer with respect to magnesium was greatly altered. These results demonstrate the function of the quaternary structure of plant acetohydroxy acid isomeroreductase in the stabilization of the tertiary structure but also in the stabilization of a high-affinity magnesium binding site. PMID- 9737853 TI - Temperature-jump and potentiometric studies on recombinant wild type and Y143F and Y254F mutants of Saccharomyces cerevisiae flavocytochrome b2: role of the driving force in intramolecular electron transfer kinetics. AB - The kinetics of intramolecular electron transfer between flavin and heme in Saccharomyces cerevisiae flavocytochrome b2 were investigated by performing potentiometric titrations and temperature-jump experiments on the recombinant wild type and Y143F and Y254F mutants. The midpoint potential of heme was determined by monitoring redox titrations spectrophotometrically, and that of semiquinone flavin/reduced flavin (Fsq/Fred) and oxidized flavin (Fox)/Fsq couples by electron paramagnetic resonance experiments at room temperature. The effects of pyruvate on the kinetic and thermodynamic parameters were also investigated. At room temperature, pH 7.0 and I = 0.1 M, the redox potential of the Fsq/Fred, Fox/Fsq, and oxidized heme/reduced heme (Hox/Hred) couples were 135, -45, and -3 mV, respectively, in the wild-type form. Although neither the mutations nor excess pyruvate did appreciably modify the potential of the heme or that of the Fsq/Fred couple, they led to variable positive shifts in the potential of the Fox/Fsq couple, thus modulating the driving force that characterizes the reduction of heme by the semiquinone in the -42 to +88 mV range. The relaxation rates measured at 16 degreesC in temperature-jump experiments were independent of the protein concentrations, with absorbance changes corresponding to the reduction of the heme. Two relaxation processes were clearly resolved in wild-type flavocytochrome b2 (1/tau1 = 1500 s-1, 1/tau2 = 200 +/- 50 s-1) and were assigned to the reactions whereby the heme is reduced by Fred and Fsq, respectively. The rate of the latter reaction was determined in the whole series of proteins. Its variation as a function of the driving force is well described by the expression obtained from electron-transfer theories, which provides evidence that the intramolecular electron transfer is not controlled by the dynamics of the protein. PMID- 9737854 TI - Calcium binding by the N-terminal cellulose-binding domain from Cellulomonas fimi beta-1,4-glucanase CenC. AB - The interaction of the N-terminal cellulose-binding domain, CBDN1, from Cellulomonas fimi beta-1,4-glucanase CenC with calcium was investigated using NMR spectroscopy and calorimetry. CBDN1 binds a single calcium ion with an equilibrium association constant of approximately 10(5) M-1 at 35 degreesC and pH 6.0. Binding is exothermic (-42 +/- 2 kJ mol-1) under these conditions and is accompanied by a small negative change in heat capacity (DeltaCp = -0.41 +/- 0.16 kJ mol-1 K-1). From an NMR line shape analysis, the rate constants for calcium association and dissociation were found to be (5 +/- 2) x 10(7) s-1 M-1 and (4.5 +/- 0.6) x 10(2) s-1, respectively. The rapid association kinetics indicate that the calcium-binding site on CBDN1 is accessible and, to the first approximation, preformed. Based on patterns of chemical shift perturbations, and structural comparisons with the Bacillus sp. 1, 3-1,4-beta-glucanases, the backbone carbonyl oxygens of Thr8, Gly30, and Asp142 and a side chain carboxyl oxygen of Asp142 are postulated to form the calcium-binding site of CBDN1. Consistent with the calcium independent affinity of CBDN1 for cellopentaose, this exposed site is located on the face of CBDN1 opposite to that forming the oligosaccharide-binding cleft. The midpoint denaturation temperature of CBDN1 is increased by approximately 8 degreesC at pH 6.0 in the presence of saturating amounts of calcium, confirming that metal ion binding is thermodynamically linked to native-state stability. PMID- 9737855 TI - Enzymological characterization of the signal-transducing uridylyltransferase/uridylyl-removing enzyme (EC 2.7.7.59) of Escherichia coli and its interaction with the PII protein. AB - The uridylyltransferase/uridylyl-removing enzyme (UTase/UR) of Escherichia coli plays an important role in the regulation of nitrogen assimilation by controlling the uridylylation state of the PII signal transduction protein (PII) in response to intracellular signals. The reversible uridylylation of PII indirectly controls the activity of PII receptors that regulate transcription from nitrogen-regulated promoters and the activity of glutamine synthetase. Here, we present a detailed analysis of the uridylyltransferase and uridylyl-removing activities and their regulation by the small molecule effectors ATP, 2-ketoglutarate, and glutamine. Several important features of enzyme mechanism and regulation were elucidated. Mg2+ appeared to be the physiologically relevant metal ion cofactor for both transferase and uridylyl-removing activities. The transferase reaction proceeded by an ordered bi-bi kinetic mechanism, with PII binding before UTP and pyrophosphate (PPi) released before PII-UMP. The uridylyl-removing reaction proceeded with rapid equilibrium binding of substrate and random release of products. Both reactions were activated by ATP and 2-ketoglutarate, which did so by binding only to PII and PII-UMP. The binding of these effectors to PII and PII UMP was characterized. Glutamine inhibited the transferase reaction by inhibiting the chemistry step, while glutamine provided nonessential mixed-type activation of the uridylyl-removing activity, lowering the apparent Km and increasing kcat. Our data were consistent with the hypothesis that all effects of glutamine are due to the binding of central complexes at a single glutamine site. By comparing the effects of the activators with their reported in vivo concentrations, we conclude that in intact cells the uridylylation state of PII is regulated mainly by the glutamine concentration and is largely independent of the 2-ketoglutarate concentration. Our kinetic data were consistent with the hypothesis that both transferase and uridylyl-removal reactions occurred at a single active center on the enzyme. PMID- 9737856 TI - Reconstitution of the signal-transduction bicyclic cascade responsible for the regulation of Ntr gene transcription in Escherichia coli. AB - Nitrogen-regulation of gene transcription in Escherichia coli results from the regulation of the phosphorylation state of the enhancer-binding transcription factor NRI (NtrC). We examined the regulation of NRI phosphorylation in a reconstituted bicyclic cascade system containing four regulatory proteins: NRI, the signal-transducing uridylyltransferase/uridylyl-removing enzyme (UTase/UR), its substrate the signal transduction protein PII, and the kinase/phosphatase NRII (NtrB), which is a PII receptor that phosphorylates and dephosphorylates NRI. In this reconstituted system, the phosphorylation state of NRI was regulated reciprocally by the small molecule effectors glutamine, which prevented the accumulation of NRI-P, and 2-ketoglutarate, which caused accumulation of NRI-P. Regulation of the bicyclic system by glutamine was exclusively due to sensation and signal-transduction by the UTase/UR-PII monocycle, which was observed to function essentially as a glutamine-sensing apparatus. In contrast, regulation of NRI phosphorylation by 2-ketoglutarate, which binds to PII, was due to direct regulation of the NRII-PII interaction and the rate of NRI-P dephosphorylation. Thus, the PII protein transduces the glutamine signal to the NRII-NRI monocycle in the form of its uridylylation state and is also the receptor of the antagonistic 2-ketoglutarate signal, which blocks the activity of unmodified PII. PMID- 9737857 TI - The regulation of Escherichia coli glutamine synthetase revisited: role of 2 ketoglutarate in the regulation of glutamine synthetase adenylylation state. AB - The regulation of Escherichia coli glutamine synthetase (GS) by reversible adenylylation has provided one of the classical paradigms for signal transduction by cyclic cascades. Yet, many mechanistic features of this regulation remain to be elucidated. We examined the regulation of GS adenylylation state in a reconstituted system containing GS, adenylyltransferase (ATase), the PII signal transduction protein that controls ATase, and the uridylyltransferase/uridylyl removing enzyme (UTase/UR), which has a role in regulating PII. In this reconstituted bicyclic cascade system, the adenylylation state of GS was regulated reciprocally by the small molecule effectors 2-ketoglutarate and glutamine at physiological effector concentrations. By examination of the individual regulatory monocycles and comparison to the bicyclic system and existing data, we could deduce that the only sensors of 2-ketoglutarate were PII and PII-UMP. At physiological conditions, we observed that the main role of 2 ketoglutarate in bringing about the deadenylylation of GS was to inhibit GS adenylylation, and this was due to the allosteric regulation of PII activity. Glutamine acted as an allosteric regulator of both ATase and UTase/UR. We also compared the regulation of GS adenylylation state to the regulation of phosphorylation state of the transcription factor NRI (NtrC) in a reconstituted bicyclic system containing NRI, the bifunctional kinase/phosphatase NRII (NtrB), PII, and the UTase/UR. This comparison indicated that, at a fixed 2-ketoglutarate concentration, the regulation of GS adenylylation state by glutamine was sharper and occurred at a higher concentration than did the regulation of NRI phosphorylation. The possible biological implications of this regulatory arrangement are discussed. PMID- 9737858 TI - Structural analysis of peptide substrates for mucin-type O-glycosylation. AB - The structures of three nine-residue peptide substrates that show differential kinetics of O-linked glycosylation catalyzed by distinct recombinant uridine diphosphate-N-acetylgalactosamine:polypeptide N-acetylgalactosaminyltransferases (GalNAc transferases) were investigated by NMR spectroscopy. A combined use of NMR data, molecular modeling techniques, and kinetic data may explain some structural features required for O-glycosylation of these substrates by two GalNAc transferases, GalNAc-T1 and GalNAc-T3. In the proposed model, the formation of an extended backbone structure at the threonine residue to be glycosylated is likely to enhance the O-glycosylation process. The segment of extended structure includes the reactive residue in a beta-like or an inverse gamma-turn conformation and flanking residues in a beta-strand conformation. The hydroxyl group of the threonine to be glycosylated is exposed to solvent, and both the amide proton and carbonyl oxygen of the peptide backbone are exposed to solvent. The exchange rate of the amide proton for the reactive threonine correlated well with substrate efficiency, leading us to hypothesize that this proton may serve as a donor for hydrogen bonding with the active site of the enzyme. The oxygens of the residue to be glycosylated and several flanking residues may also be involved in a set of hydrogen bonds with the GalNAc-T1 and T3 transferases. PMID- 9737859 TI - Stability of recombinant yeast protoporphyrinogen oxidase: effects of diphenyl ether-type herbicides and diphenyleneiodonium. AB - Protoporphyrinogen oxidase catalyzes the oxygen-dependent aromatization of protoporphyrinogen IX to protoporphyrin IX and is the molecular target of diphenyl ether-type herbicides. Structural features of yeast protoporphyrinogen oxidase were assessed by circular dichroism studies on the enzyme purified from E. coli cells engineered to overproduce the protein. Coexpression of the bacterial gene ArgU that encodes tRNAAGA,AGG and a low induction temperature for protein synthesis were critical for producing protoporphyrinogen oxidase as a native, active, membrane-bound flavoprotein. The secondary structure of the protoporphyrinogen oxidase was 40.0 +/- 1. 5% alpha helix, 23.5 +/- 2.5% beta sheet, 18.0 +/- 2.0% beta turn, and 18.5 +/- 2.5% random-coil. Purified protoporphyrinogen oxidase appeared to be a monomeric protein that was relatively heat-labile (Tm of 44 +/- 0.5 degreesC). Acifluorfen, a potent inhibitor that competes with the tetrapyrrole substrate, and to a lower extent FAD, the cofactor of the enzyme, protected the protein from thermal denaturation, raising the Tm to 50.5 +/- 0.5 degreesC (acifluorfen) and 46.5 +/- 0.5 degreesC (FAD). However, diphenyleneiodonium, a slow tight-binding inhibitor that competes with dioxygen, did not protect the enzyme from heat denaturation. Acifluorfen binding to the protein increased the activation energy for the denaturation from 15 to 80 kJ.mol 1. The unfolding of the protein was a two-step process, with an initial fast reversible unfolding of the native protein followed by slow aggregation of the unfolded monomers. Functional analysis indicated that heat denaturation caused a loss of enzyme activity and of the specific binding of radiolabeled inhibitor. Both processes occurred in a biphasic manner, with a transition temperature of 45 degreesC. PMID- 9737861 TI - The delta-subunit of pyruvate ferredoxin oxidoreductase from Pyrococcus furiosus is a redox-active, iron-sulfur protein: evidence for an ancestral relationship with 8Fe-type ferredoxins. AB - Pyruvate ferredoxin oxidoreductase (POR) from the hyperthermophilic archaeon Pyrococcus furiosus (Pf) catalyzes the final oxidative step in carbohydrate fermentation in which pyruvate is oxidized to acetyl-CoA and CO2, coupled to the reduction of ferredoxin (Fd). POR is composed of two 'catalytic units' of molecular mass approximately 120 kDa. Each unit consists of four subunits, alpha beta gamma delta, with masses of approximately 44, 36, 20, and 12 kDa, respectively, and contains at least two [4Fe-4S] clusters. The precise mechanism of catalysis and the role of the individual subunits are not known. The gene encoding the delta-subunit of Pf POR has been expressed in E. coli, and the protein was purified after reconstitution with iron and sulfide. The reconstituted delta-subunit (recPOR-delta) is monomeric with a mass of 11 879 +/- 1.2 Da as determined by mass spectrometry, in agreement with that predicted from the gene sequence. Purified recPOR-delta contains 8 Fe mol/mol and remained intact when incubated at 85 degreesC for 2 h, as judged by its visible absorption properties. The reduced form of the protein exhibited an EPR spectrum characteristic of two, spin-spin interacting [4Fe-4S]1+ clusters. When compared with the EPR properties of the reduced holoenzyme, the latter was shown to contain a third [4Fe-4S]1+ cluster in addition to the two within the delta subunit. The reduction potential of the two 4Fe clusters in isolated recPOR-delta (-403 +/- 8 mV at pH 8.0 and 24 degreesC) decreased linearly with temperature ( 1.55 mV/ degreesC) up to 82 degreesC. RecPOR-delta replaced Pf Fd as an in vitro electron carrier for two oxidoreductases from Pf, POR and Fd:NADP oxidoreductase, and the POR holoenzyme displayed a higher apparent affinity for its own subunit (apparent Km = 1.0 microM at 80 degreesC) than for Fd (apparent Km = 4.4 microM). The molecular and spectroscopic properties and amino acid sequence of the isolated delta-subunit suggest that it evolved from an 8Fe-type Fd by the addition of approximately 40 residues at the N-terminus, and that this extension enabled it to interact with additional subunits within POR. PMID- 9737860 TI - Delta T 14/Delta D 15 Azotobacter vinelandii ferredoxin I: creation of a new CysXXCysXXCys motif that ligates a [4Fe-4S] cluster. AB - In clostridial-type ferredoxins, each of the two [4Fe-4S]2+/+ clusters receives three of its four ligands from a CysXXCysXXCys motif. Azotobacter vinelandii ferredoxin I (AvFdI) is a seven-iron ferredoxin that contains one [4Fe-4S]2+/+ cluster and one [3Fe-4S]+/0 cluster. During the evolution of the 7Fe azotobacter type ferredoxins from the 8Fe clostridial-type ferredoxins, one of the two motifs present changed to a CysXXCysXXXXCys motif, resulting in the inability to form a 4Fe cluster and the appearance of a 3Fe cluster in that position. In a previous study, we were unsuccessful in using structure as a guide in designing a 4Fe cluster in the 3Fe cluster position of AvFdI. In this study, we have reversed part of the evolutionary process by deleting two residues between the second and third cysteines. UV/Vis, CD, and EPR spectroscopies and direct electrochemical studies of the purified protein reveal that this DeltaT14/DeltaD15 FdI variant is an 8Fe protein containing two [4Fe-4S]2+/+ clusters with reduction potentials of 466 and -612 mV versus SHE. Whole-cell EPR shows that the protein is present as an 8Fe protein in vivo. These data strongly suggest that it is the sequence motif rather than the exact sequence or the structure that is critical for the assembly of a 4Fe cluster in that region of the protein. The new oxygen-sensitive 4Fe cluster was converted in partial yield to a 3Fe cluster. In known ferredoxins and enzymes that contain reversibly interconvertible [4Fe-4S]2+/+ and [3Fe-4S]+/0 clusters, the 3Fe form always has a reduction potential ca. 200 mV more positive than the 4Fe cluster in the same position. In contrast, for DeltaT14/DeltaD15 FdI, the 3Fe and 4Fe clusters in the same location have extremely similar reduction potentials. PMID- 9737862 TI - Identification of a meander region proline residue critical for heme binding to cytochrome P450: implications for the catalytic function of human CYP4B1. AB - Alignment of xenobiotic-metabolizing P450 protein sequences highlights an invariant proline residue in the meander region two amino acids N-terminal to the distal arginine of the putative ERR triad thought to be important for heme binding. This occurs as a serine in the sequences derived from human CYP4B1 gDNA and both human lung and placental CYP4B1 cDNAs. Reversion of this serine to the conserved proline residue (Ser427 --> Pro) by site-directed mutagenesis conferred the ability to incorporate heme on the human placental enzyme. Mutation of the corresponding proline in rabbit CYP4B1 (Pro422 --> Ser) abolished heme incorporation. Membrane preparations of human CYP4B1(Pro) and rabbit CYP4B1(Pro), but not the corresponding CYP4B1(Ser) variants, supported lauric acid hydroxylation preferentially at the omega-position. Purified, reconstituted human CYP4B1(Pro) and rabbit CYP4B1(Pro) formed 12-hydroxylauric acid at rates of 17-21 min-1, and both enzymes were also C-8 to C-10 fatty acid omega-hydroxylases preferentially, with total rates of hydroxylation decreasing in the order C-12 > C-10 > C-9 > C-8. Finally, neither human nor rabbit CYP4B1(Pro) formed detectable levels of any hydroxylated testosterone metabolites. Therefore, the presence of a consensus Pro-X-Arg motif is critical for incorporation of the heme prosthetic group in human and rabbit CYP4B1 proteins expressed in insect cells. Native human CYP4B1, expressed in vivo, is likely to be functionally impaired if Pro427 is required for holoenzyme expression in mammalian cells. PMID- 9737863 TI - Interactions among P450 enzymes when combined in reconstituted systems: formation of a 2B4-1A2 complex with a high affinity for NADPH-cytochrome P450 reductase. AB - The purpose of this study is to characterize the interactions among P450 1A2, P450 2B4, and P450 reductase in mixed reconstituted systems. Previously, our laboratory demonstrated that in the presence of certain substrates, 1A2 can influence the catalytic characteristics of 2B4 [Cawley et al. (1995) Biochemistry 34, 1244-1247]. The goal of the current study is to distinguish between two models to explain these interactions: one model where substrate increases the affinity of one P450 enzyme for the reductase, and another model where substrate increases the affinity of one P450 for the reductase through the formation of a 1A2-2B4 complex. According to this model, the 1A2 moiety of 1A2-2B4 forms a high affinity complex with reductase. Reductase, 1A2, and 2B4 were reconstituted with dilauroylphosphatidylcholine, and the effect of reductase concentration on 7 pentoxyresorufin-O-dealkylation was examined with 2B4-reductase and 1A2-reductase binary systems, and in ternary systems containing different 2B4:1A2 ratios. At subsaturating [reductase], there was a dramatic inhibition of the 2B4-dependent activity in the ternary system as compared with the binary systems. These results are consistent with the formation of a ternary (reductase-1A2-2B4) complex where the reductase is bound specifically to 1A2. At higher reductase concentrations where the reductase-binding sites on 1A2 become saturated, the results are consistent with the formation of a quaternary complex in which reductase binds to both P450 enzymes (reductase-1A2-2B4-reductase). Analogous experiments using the 1A2-preferred substrate 7-ethoxyresorufin showed a stimulation of 7 ethoxyresorufin-O-deethylation in the mixed reconstituted system, demonstrating that the high-affinity 2B4-1A2-reductase complex was functionally active and not merely an inhibitory complex. PMID- 9737864 TI - Membrane insertion of cytochrome P450 1A2 promoted by anionic phospholipids. AB - The role of phospholipids in the membrane binding and subsequent insertion of the microsomal protein rabbit cytochrome P450 (P450) 1A2 into phospholipid bilayers was investigated. The insertion of P450 1A2 into phospholipid bilayers was measured by the quenching of Trp fluorescence of P450 1A2 by pyrene and brominated and doxyl-labeled phospholipids. When the phosphatidylcholine (PC) matrix was replaced with acidic phospholipids [phosphatidic acid (PA), phosphatidylserine, and phosphatidylinositol] and phosphatidylethanolamine (PE), the extent of insertion into lipid bilayers was strictly dependent on the type of acidic phospholipids. All anionic phospholipids caused the penetration of P450 1A2 into lipid bilayers, but PA was the most efficient in facilitating deep penetration of P450 1A2 into bilayers. On the other hand, binding of P450 1A2 to liposomes was increased by acidic phospholipids to the same degree regardless of the type of acidic phospholipids. PE was found to act as an inert matrix phospholipid, similar to PC, as it exerted very little effect on the insertion of P450 1A2 into lipid bilayers and the binding of P450 1A2 to membranes. It was also found that the phospholipid-dependent membrane insertion of P450 1A2 was associated with altered enzyme activity, increased alpha-helix content, and increased Trp fluorescence of P450 1A2. These results indicate that negative charges on the acidic phospholipids are important for the initial binding of P450 1A2 to membranes, but the penetration of P450 1A2 into lipid bilayers is regulated by the type of acidic phospholipids, and that phospholipid-dependent insertion of P450 1A2 is accompanied by a structural change of P450 1A2. PMID- 9737865 TI - Apolipoprotein B-100 conformation and particle surface charge in human LDL subspecies: implication for LDL receptor interaction. AB - The plasma low-density lipoprotein (LDL) profile in coronary artery disease patients is characterized by a predominance of small, dense LDL. Small, dense LDL exhibit both high susceptibility to oxidation and low binding affinity for the LDL receptor, suggesting that these particles may be of elevated atherogenic potential. Here we examine whether the variation in biological function is due to differences in apo B-100 conformation that alter the interaction with the cellular LDL receptor. The microenvironments (pKa) of Lys residues in apo B-100 in small, dense, intermediate, and light human LDL subspecies have been compared by 13C NMR, and the net surface charge of these particles has been characterized. Relative to the total LDL fraction, small, dense, and light LDL subspecies have a decreased number of pKa 8.9 Lys, while intermediate density LDL has a consistently higher number of pKa 8.9 Lys. It follows that differences in protein conformation, as reflected in the Lys microenvironments, exist in the different LDL subspecies. Electrophoretic mobility measurements revealed that the light LDL subfractions exhibit a surface charge at pH 8.6 that is from -26 to -34e more negative than the intermediate density LDL subfraction. For the small, dense LDL particles the increments in negative charge range from -7 to -17e relative to the intermediate density LDL subfraction. These results suggest that differences in the conformation of apo B-100 and surface charge between LDL subspecies are major determinants of their catabolic fate. The lower number of pKa 8.9 Lys leads to a reduction in binding of small, dense, and light LDL to the cellular LDL receptor and prolongs their plasma residence time, thereby elevating the atherogenicity of these particles. These data support the proposal that the intermediate LDL subspecies constitute the optimal ligand for the LDL receptor among human LDL particle subpopulations. PMID- 9737866 TI - Liposome-cell interactions in vitro: effect of liposome surface charge on the binding and endocytosis of conventional and sterically stabilized liposomes. AB - The cellular uptake of liposomes is generally believed to be mediated by adsorption of liposomes onto the cell surface and subsequent endocytosis. This report examines the effect of liposome surface charge on liposomal binding and endocytosis in two different cell lines: a human ovarian carcinoma cell line (HeLa) and a murine derived mononuclear macrophage cell line (J774). The large unilamellar liposomes were composed of 1, 2-dioleolyl-sn-glycero-3 phosphatidylcholine with and without the addition of either a positively charged lipid, 1, 2-dioleoyl-3-dimethylammonium propanediol (DODAP), or a negatively charged lipid, 1,2-dioleolyl-sn-glycero-3-phosphatidylserine. In some experiments 5 mol % of the anionic PEG2000-PE or a neutral PEG lipid of the same molecular weight was added. HeLa cells were found to endocytose positively charged liposomes to a greater extent than either neutral or negatively charged liposomes. This preference was not lipid-specific since inclusion of a cationic cyanine dye, DiIC18(3), to impart positive charge in place of DODAP resulted in a similar extent of endocytosis. In contrast the extent of liposome interaction with J774 cells was greater for both cationic and anionic liposomes than for neutral liposomes. The greater uptake of positively charged liposomes by HeLa cells was also observed with sterically stabilized liposomes (PEG liposomes). Although the overall amount of endocytosis for all the PEG liposomes examined was attenuated relative to conventional liposomes, the extent of endocytosis was greatest for positively charged PEG liposomes, whereas negatively charged PEG2000 PE liposomes were hardly endocytosed by the HeLa cells. Incorporation of a neutral PEG lipid into liposomes permits the independent variation of liposome steric and electrostatic effects in a manner that may allow interactions with cells of the reticuloendothelial system to be minimized, yet permit strong interactions between liposomes and proliferating cells. PMID- 9737867 TI - Transport of arachidonic acid across the neutrophil plasma membrane via a protein facilitated mechanism. AB - Arachidonic acid is the rate-limiting substrate in the biosynthesis of leukotrienes in activated neutrophils. Liberation of arachidonate from intracellular membranes and uptake of exogenous arachidonate are the two principal mechanisms by which the cell can increase the level of this substrate. We investigated arachidonate uptake and export by using intact polymorphonuclear neutrophils and inside-out plasma membrane vesicles thereof. Here we show that the cellular uptake of arachidonate is energy dependent with an energy of activation (EA) of 10.0 kcal/mol and half-saturated at an arachidonate concentration of 4.8 nmol/mg of cell protein. Protein-facilitated transport of arachidonate across the plasma membrane in either direction is sensitive to proteases, chemical protein modifying reagents, anion transport inhibitors, and, most notably, toward several structurally unrelated leukotriene B4 receptor antagonists with IC50 values in the range of 16-44 microM. The inhibitors did not inhibit the diffusional uptake of methyl arachidonate into neutrophils and inside out plasma membrane vesicles, indicating that a transport protein is required for the rapid uptake of the free acid but not for the uptake of the ester. Other long chain fatty acids did compete with the uptake of arachidonate in both assay systems, whereas leukotriene B4 did not. This study documents a novel protein facilitated transport mechanism for arachidonate in neutrophils, potentially involved in transcellular eicosanoid biosynthesis and sPLA2-mediated arachidonate signaling in neutrophils. PMID- 9737868 TI - N,N-Dimethylsphingosine is a potent competitive inhibitor of sphingosine kinase but not of protein kinase C: modulation of cellular levels of sphingosine 1 phosphate and ceramide. AB - Sphingosine 1-phosphate (SPP), a lipid second messenger formed by the action of sphingosine kinase, has been implicated in regulating diverse biological processes, including growth, survival, and differentiation. N,N Dimethylsphingosine (DMS) inhibits sphingosine kinase and has been used to investigate the biological roles of SPP; however, little is known of the mechanism of inhibition of sphingosine kinase by DMS. In addition, DMS has been shown to inhibit protein kinase C in vitro. Here we report that DMS is a competitive inhibitor of sphingosine kinase from U937 monoblastic leukemia cells, Swiss 3T3 fibroblasts, and PC12 pheochromocytoma cells. DMS decreases basal levels of SPP and prevents increases in SPP in response to physiological stimuli known to activate sphingosine kinase. DMS also effectively increases cellular levels of ceramide in a variety of cell types, and resetting of the ceramide/SPP rheostat may account for the pro-apoptotic effects of DMS. Moreover, DMS, at concentrations which effectively inhibit sphingosine kinase, has no effect on protein kinase C activity or its membrane translocation. Thus, DMS acts as a specific competitive inhibitor of sphingosine kinase in diverse cell types and is a useful tool to elucidate the role of SPP as an intracellular second messenger. PMID- 9737869 TI - Functional analysis of conserved domains in the phosphotyrosyl phosphatase activator. Molecular cloning of the homologues from Drosophila melanogaster and Saccharomyces cerevisiae. AB - Phosphotyrosyl phosphatase activator (PTPA), a 37 kDa cytosolic protein that specifically activates the phosphotyrosyl phosphatase activity of the dimeric form of PP2A, was cloned from Drosophila melanogaster and Saccharomyces cerevisiae. Sequence alignment of PTPA from yeast to human revealed highly conserved regions including the type B fragment of the putative PTPA ATP binding site. We generated PTPA deletion mutants of these conserved regions as well as point mutations within regions that were suggested to be functionally important. The recombinant proteins were expressed in E. coli and subsequently purified. Activity measurements, linked with immunological detection, revealed that most of the well-conserved regions are essential for PTPA activity. However, neither the type A fragment of the putative ATP binding site nor the cysteine-rich region, present in all but the Drosophila and yeast homologues, appeared to be essential for PTPA activity. Moreover, we observed that PTPA truncated at glycine266 behaves as a dominant negative mutant since it is inhibitory to the wild-type PTPA. PMID- 9737870 TI - Kinetic studies of FR-1, a growth factor-inducible aldo-keto reductase. AB - Murine fibroblasts cultured in the presence of fibroblast growth factor-1 express relatively high levels of FR-1, a approximately 36 kDa protein related to the aldo-keto reductase superfamily [Donohue, P. J., Alberts, G. F., Hampton, B. S., Winkles, J. A. (1994) J. Biol. Chem. 269, 8604-8609]. While the crystal structure of FR-1 shows striking homology with human aldose reductase [Wilson, D. K., Nakano, T., Petrash, J. M., Quiocho, F. A. (1995) Biochemistry 34, 14323-14330], an enzyme linked to the pathogenesis of diabetic complications, the physiological role of FR-1 is not known. We show that FR-1 is capable of reducing a broad range of aromatic and aliphatic aldehydes, including the abundant and highly reactive lipid-derived aldehyde 4-hydroxy-2-nonenal (HNE; Km approximately 9 microM). However, in the absence of coenzyme, HNE caused a time-dependent inactivation of FR-1. Results from electrospray ionization-mass spectrometry and Edman degradation of peptides derived from HNE-modified FR-1 were consistent with formation of a Michael adduct at Cys298. This was confirmed with a C298S mutant, which was resistant to HNE-induced inactivation. Since steady-state Km values determined with alkanals, alpha,beta-unsaturated alkenals, alkadienals, and 4 hydroxyalkenals fall within their physiological concentrations, lipid-derived aldehydes appear to be potential in vivo substrates for FR-1. PMID- 9737871 TI - Factors influencing the dimer to monomer transition of an antibody single-chain Fv fragment. AB - Antibody single-chain Fv (scFv) fragments are able to form dimers under certain conditions, and the extent of dimerization appears to depend on linker length, antibody sequence, and external factors. We analyzed the factors influencing dimer-monomer equilibrium as well as the rate of interconversion, using the scFv McPC603 as a model system. In this molecule, the stability of the VH-VL interaction can be conveniently varied by adjusting the ionic strength (because of its influence on the hydrophobic effect), by pH (presumably because of the presence of titratable groups in the interface), and by the presence or absence of the antigen phosphorylcholine, which can be rapidly removed due to its very fast off-rate. It was found that the monomer is the thermodynamically stable form with linkers of 15 and 25 amino acids length under all conditions tested (35 microM or less). The dimer is initially formed in periplasmic expression, presumably by domain swapping, and can be trapped by all factors which stabilize the VH-VL interface, such as the presence of the antigen, high ionic strength, and pH below 7.5. Under all other conditions, it converts to the monomer. Predominantly monomer is obtained during in vitro folding. Monomer is stabilized against dimerization at very high concentrations by the same factors which stabilize the VH-VL interaction. These results should be helpful in producing molecules with defined oligomerization states. PMID- 9737873 TI - Identifying RNA minor groove tertiary contacts by nucleotide analogue interference mapping with N2-methylguanosine. AB - Nucleotide analogue interference mapping (NAIM) is a general biochemical method that rapidly identifies the chemical groups important for RNA function. In principle, NAIM can be extended to any nucleotide that can be incorporated into an in vitro transcript by an RNA polymerase. Here we report the synthesis of 5'-O (1-thio)-N2-methylguanosine triphosphate (m2GalphaS) and its incorporation into two reverse splicing forms of the Tetrahymena group I intron using a mutant form of T7 RNA polymerase. This analogue replaces one proton of the N2 exocyclic amine with a methyl group, but is as stable as guanosine (G) for secondary structure formation. We have identified three sites of m2GalphaS interference within the Tetrahymena intron: G22, G212, and G303. All three of these guanosine residues are known to utilize their exocyclic amino groups to participate in tertiary hydrogen bonds within the ribozyme structure. Unlike the interference pattern with the phosphorothioate of inosine (IalphaS, an analogue that deletes the N2 amine of G), m2GalphaS substitution did not cause interference at positions attributable to secondary structural stability effects. Given that the RNA minor groove is likely to be widely used for helix packing, m2GalphaS provides an especially valuable reagent to identify RNA minor groove tertiary contacts in less well-characterized RNAs. PMID- 9737872 TI - Formation of proteasome-PA700 complexes directly correlates with activation of peptidase activity. AB - The proteolytic activity of the eukaryotic 20S proteasome is stimulated by a multisubunit activator, PA700, which forms both 1:1 and 2:1 complexes with the proteasome. Formation of the complexes is enhanced by an additional protein assembly called modulator, which also stimulates the enzymatic activity of the proteasome only in the presence of PA700. Here we show that the binding of PA700 to the proteasome is cooperative, as is the activation of the proteasome's intrinsic peptidase activity. Modulator increases the extent of complex formation and peptidase activation, while preserving the cooperative kinetics. Furthermore, the increase in activity is not linear with the number of PA700 assemblies bound to the proteasome, but rather with the number of proteasome-PA700 complexes, regardless of the PA700:proteasome stoichiometry. Hence the stimulation of peptidase activity is fully (or almost fully) effected by the binding of a single PA700 to the 20S proteasome. The stimulation of peptidase by modulator is explained entirely by the increased number of proteasome-PA700 complexes formed in its presence, rather than by any substantial direct stimulation of catalysis. These observations are consistent with a model in which PA700, either alone or assisted by modulator, promotes conformational changes in the proteasome that activate the catalytic sites and/or facilitate access of peptide substrates to these sites. PMID- 9737874 TI - In vivo expression of a variant human U6 RNA from a unique, internal promoter. AB - We previously isolated a variant of the human U6 small nuclear RNA gene (87U6) and demonstrated that transcription of this gene is controlled by a novel internal promoter. It has now been shown that two blocks of sequence within the coding region are both necessary and sufficient to direct expression of 87U6 in transcription assays performed in vitro. In addition, 87U6 is expressed in vivo and can assemble into snRNP complexes. Specific primer extension assays on total RNA from HeLa cells shows that 87U6 RNA is present in these cells. Also, microinjection of plasmid encoded 87U6 genes into Xenopus laevis oocyte nuclei results in the expression of this variant RNA. Immunoprecipitation with anti-Sm antibodies suggests that 87U6 RNA assembles into a snRNP particle with U4 snRNA. Finally, the variant snRNA is capped with the U6 specific gamma-monomethyl phosphate cap when incubated in HeLa extracts. These data suggest that 87U6 RNA may function in the splicing process, in a manner similar to the wild-type U6 RNA. The recent observations of a minor class of mRNA introns that are spliced by a distinct collection of snRNP particles suggest an important role for variant snRNAs in the splicing of transcripts with alternative splice junctions. PMID- 9737875 TI - Stabilization of intramolecular triple/single-strand structure by cationic peptides. AB - For better comprehension of possible physiological roles of triple-helical DNA structures, it is important to understand if the proteins can stabilize intramolecular triplex (H-DNA). One plausible mode of stabilization is through the neutralization of electrostatic repulsion of negatively charged phosphates in the three DNA strands by positively charged arginine and lysine residues of a bound protein. To gain an insight into interactions between H-DNA and cationic protein domains, we examined the effect of Lys- and Arg-rich oligopeptides on the B-DNA to H-DNA transition. These oligopeptides as well as another type of polycation, spermine, shifted the equilibrium toward H-DNA. These polycations introduced little change in DNA superhelicity, so that an increase in torsional stress was not responsible for facilitated H-DNA formation. Competing influences of polycations and monovalent cations suggest a significant involvement of electrostatic interactions in H-DNA stabilization. The Arg-rich peptides are more effective in H-DNA stabilization than the Lys-rich ones. However, as inferred from experiments on intermolecular complexes, this is not due to a better stabilization of triple helix or destabilization of double helix. It is possible that Arg-rich peptides interact with the unpaired single strand in H-DNA and stabilize its unpaired conformation. PMID- 9737876 TI - Structure of d(GT)n.d(GA)n sequences: formation of parallel stranded duplex DNA. AB - Alternating polypurine sequences exhibit remarkable polymorphism. In this study, we report that dGA.dGT sequences form parallel stranded duplex DNA at neutral pH. Using two model hairpins, 3'-d(GT)3-5'5'-T4(AG)3-3' (I) and 3'-d(GT)4-5'5' T4(AG)4-3' (II), containing 5'5' linkages which direct parallel strand formation, we systematically explored the spectroscopic and thermodynamic properties of parallel stranded d(GA)n.d(GT)n. The parallel stranded hairpins are remarkably stable structures with TM's of 41.5 and 47.5 degreesC (in 0.4 M NaCl) for the shorter and longer hairpins, respectively. The van't Hoff enthalpies of 80.7 and 114 kJ mol-1 are relatively low but are comparable to a parallel stranded d(GA)n duplex. On the basis of the spectroscopic and electrophoretic characteristics, we conclude that parallel strand formation is not restricted to hairpin systems, but also readily occurs in unconstrained dimeric duplexes with the appropriate sequence homologies. Both melting curves and electrophoretic analyses of parallel stranded heteroduplexes in which the sequence enforces specific base pairing demonstrate that G-G and A-T base pairs are formed in d(GA)n.d(GT)n segments. PMID- 9737877 TI - Fluorescence correlation spectroscopy of enzymatic DNA polymerization. AB - We show that fluorescence correlation spectroscopy (FCS) can be used as a reliable, simple, and fast tool for detecting products of the polymerase chain reaction (PCR). By use of autocorrelation experiments, it is demonstrated that fluorescent 217-bp DNA fragments can be detected at very low initial ss M13mp18(+) DNA and tetramethylrhodamine-4-dUTP concentrations and that these polymers are cleaved by the chosen restriction enzymes. A FCS calibration curve is presented, where the translational diffusion times of different size DNA fragments are plotted versus the number of base pairs they contain. At zero and very low template concentrations a large "background" species emerges, which is a reflection of the experimental conditions chosen and the extremely high sensitivity of FCS. The relative amount of nonspecific product formation is less than 1%. The ease by which a FCS measurement can be performed (a few minutes at most) also enables the technique to be used as an effective screening method. PMID- 9737878 TI - 1H nuclear magnetic resonance study on equilibrium between two four-stranded solution conformations of short d(CnT). AB - NMR analysis of d(C4T) showed the slow exchange between two distinct tetramers (each fully symmetric) in solution. For one tetramer, NOE cross-peak patterns characteristic of an i-motif structure (H1'-H1' and H6-H1'/H1'-H6) were observed between C1 and T5, indicating that this tetramer takes a completely intercalated conformation where the T5 residue is stacked on the C1.C1(+) pair of the other duplex (S-form). The other was found to be a tetramer in which one of the duplexes is shifted by one nucleotide unit (R-form), resulting in nonstacking 3' end thymidine residues and an equal number of stacked C.C+ pairs to that of the S form. The same spectral features were observed for d(C3T) but neither for d(TC3) nor d(TC4), indicative of the critical role of the position of the thymidine residue in the tetrad isomerization. From NMR denaturation profiles, the S-forms were found to be more stable than the R-forms, and the linear relationship between the logarithm of the equilibrium constant (K = [tetramer]/[single]4) and the inverse of temperature (1/T) was confirmed for both forms, indicating conformity to the two-state transition model. Both enthalpy and entropy values of the formation of the S-form from four single strands were more negative than those of the R-form. The enthalpy term should contribute to the stabilization of the S-forms at low temperatures. The difference of the free energy values [DeltaG degrees(S-form) - DeltaG degrees(R-form)] was found to be -2.1 and -2.7 kJ.mol-1 at 20 degreesC for d(C4T) and d(C3T), respectively, explaining the higher stability of the S-forms. With increasing temperature, these two topologies were found to comparably exist at equilibrium in solution with slow exchange via dissociation to the single strands. A biological role of this topological isomerization is also suggested. PMID- 9737879 TI - Dual regulation of the skeletal muscle ryanodine receptor by triadin and calsequestrin. AB - Triadin, a calsequestrin-anchoring transmembrane protein of the sarcoplasmic reticulum (SR), was successfully purified from the heavy fraction of SR (HSR) of rabbit skeletal muscle with an anti-triadin immunoaffinity column. Since depletion of triadin from solubilized HSR with the column increased the [3H]ryanodine binding activity, we tested a possibility of triadin for a negative regulator of the ryanodine receptor/Ca2+ release channel (RyR). Purified triadin not only inhibited [3H]ryanodine binding to the solubilized HSR but also reduced openings of purified RyR incorporated into the planar lipid bilayers. On the other hand, calsequestrin, an endogenous activator of RyR [Kawasaki and Kasai (1994) Biochem. Biophys. Res. Commun. 199, 1120-1127; Ohkura et al. (1995) Can. J. Physiol. Pharmacol. 73, 1181-1185] potentiated [3H]ryanodine binding to the solubilized HSR. Ca2+ dependency of [3H]ryanodine binding to the solubilized HSR was reduced by triadin, whereas that was enhanced by calsequestrin. Interestingly, [3H]ryanodine binding to the solubilized HSR potentiated by calsequestrin was reduced by triadin. Immunostaining with anti-triadin antibody proved that calsequestrin inhibited the formation of oligomeric structure of triadin. These results suggest that triadin inhibits the RyR activity and that RyR is regulated by both triadin and calsequestrin, probably through an interaction between them. In this paper, triadin has been first demonstrated to have an inhibitory role in the regulatory mechanism of the RyR. PMID- 9737880 TI - Microtubule dynamic instability does not result from stabilization of microtubules by tubulin-GDP-Pi subunits. AB - The proposal that microtubule dynamic instability results from stabilization of microtubule ends by tubulin-GDP-Pi subunits (where Pi is inorganic phosphate) [Melki et al. (1996) Biochemistry 35, 12038] was based on studies of GTP hydrolysis and microtubule assembly that showed that tubulin-GDP-Pi subunits can transiently accumulate at microtubule ends. There is no direct evidence that GDP Pi-subunits can stabilize microtubules under conditions where dynamic instability is observed and this has been inferred from the observation that tubulin-GDP-BeFn subunits stabilize microtubules. To test if tubulin-GDP-Pi stabilizes microtubules we sought evidence for a synergism between the effect of Pi and BeFn. We found, however, that Pi antagonizes the effect of BeFn by displacing it from tubulin subunits. The alternate mechanism in which Pi inhibits BeFn stabilization of microtubules by displacing fluoride from beryllium was ruled out from the 9Be and 19F NMR spectra in the presence and absence of Pi. Further evidence that tubulin-GDP-BeFn is not an analogue of tubulin-GDP-Pi and that tubulin-GDP-Pi is not responsible for maintaining the growth phase in microtubules manifesting dynamic instability was provided by our observation that Pi did not decrease the disassembly rate under conditions where tubulin-GDP-Pi subunits are expected to have formed. Results showing that BeFn binds randomly to subunits in microtubules provided evidence that Pi dissociation from the tubulin GDP-Pi intermediate formed during GTP hydrolysis occurs randomly rather than processively starting at the growing microtubule tip. PMID- 9737881 TI - Mapping human interferon-alpha (IFN-alpha 2) binding determinants of the type I interferon receptor subunit IFNAR-1 with human/bovine IFNAR-1 chimeras. AB - Type I interferons bind to a common receptor (IFNAR), composed of two transmembrane polypeptides, IFNAR-1 and IFNAR-2. Although human IFNAR-1 has a weak intrinsic affinity for human Type I interferons (IFNs), bovine IFNAR-1 binds human Type I IFNs with moderate (nM) affinity, and can be conveniently used to investigate the regions of IFNAR-1 involved in ligand binding. We have constructed 14 bovine/human IFNAR-1 chimeras by exchanging homologous subdomains in the extracellular portion of the receptor. These chimeras were expressed at very high levels on COS cells, and their ability to bind HuIFN-alpha2 was measured. No single bovine subdomain substituted into human IFNAR-1 could confer moderate-affinity ligand binding on the resulting chimera. Simultaneous substitution of bovine IFNAR-1 subdomains 2 and 3 for the homologous human subdomains resulted in a dramatic increase in the binding of IFN-alpha2, suggesting that critical determinants for moderate-affinity ligand binding by BoIFNAR-1 reside in these two subdomains. Bovine subdomains 1 and/or 4 each further enhanced IFN-alpha2 binding in the presence of bovine subdomains 2 and 3. Thus, the binding interactions of BoIFNAR-1 with IFNs appears to be more complex than that of other class II cytokine receptors with their ligands. PMID- 9737882 TI - Thermodynamics and kinetics of the reaction of a single-chain antibody fragment (scFv) with the leucine zipper domain of transcription factor GCN4. AB - Single-chain Fv (scFv) fragments of antibodies have become important analytical and therapeutic tools in biology and medicine. The reaction of scFv fragments has not been well-characterized with respect to the energetics and kinetics of antigen binding. This paper describes the thermodynamic and kinetic behavior of the high-affinity scFv fragment SW1 directed against the dimeric leucine zipper domain of the yeast transcription factor GCN4. The scFv fragment was selected by the phage display technique from the immune repertoire of a mouse that had been immunized with the leucine zipper domain of GCN4. The scFv fragment was produced in high yield in Escherichia coli inclusion bodies and refolded from the denatured state. Differential scanning calorimetry showed that SW1 was stable up to about 50 degreesC, but the subsequent thermal denaturation was irreversible (Tm approximately 68 degreesC). The scFv fragment specifically recognized the dimeric leucine zipper conformation. Two scFv fragments bound to the GCN4 dimer to form the complex (scFv)2-GCN4. Because of its repetitive structure, the rod shaped GCN4 leucine zipper may present two similar epitopes for the scFv fragment. Surprisingly, the binding reaction was highly cooperative, that is, the species (scFv)2-GCN4 dominated over scFv-GCN4 even in the presence of a large excess of the antigen GCN4. It is speculated that cooperativity resulted from direct interaction between the two GCN4-bound scFv fragments. At 25 degreesC, the average binding enthalpy for a scFv fragment was favorable (-61 kJ mol-1), the entropy change was unfavorable, and the change in heat capacity was -1.27 +/- 0.14 kJ mol-1 K-1. As a result of enthalpy-entropy compensation, the free binding energy was virtually independent of temperature in the physiological temperature range. Antigen binding in solution could be described by a single-exponential reaction with an apparent rate constant of 1 x 10(6) M-1 s-1. Binding followed in a biosensor with the dimeric GCN4 coupled to the surface of the metal oxide sensor chip was 20 times slower. PMID- 9737883 TI - NMR study suggests a major role for Arg111 in maintaining the structure and dynamical properties of type II human cellular retinoic acid binding protein. AB - The solution structure of a site-directed mutant of type-II human cellular retinoic acid binding protein (CRABPII) with Arg111 replaced by methionine (R111M) has been determined by NMR spectroscopy. The sequential assignments of the 1H and 15N resonances of apo-R111M were established by multinuclear multidimensional NMR. The solution structure was calculated from 2302 distance restraints and 77 phi dihedral restraints derived from the NMR data. The root mean-square deviation of the ensemble of 28 refined conformers that represent the structure from the mean coordinate set derived from them was 0.54 +/- 0.26 and 0.98 +/- 0.23 A for the backbone atoms and all heavy atoms, respectively. The solution structure of apo-R111M is similar to that of wild-type apo-CRABPII. However, there are significant conformational differences between the two proteins, localized mainly to three segments (Leu19-Ala36, Glu73-Cys81, and Leu99 Pro105) clustered around the ligand entrance more than 17 A away from the point mutation. In apo-R111M, all the three segments move toward the center of the ligand entrance so that the opening of the ligand-binding pocket in apo-R111M is much smaller than that in wild-type apo-CRABPII. Furthermore, the ligand-binding pocket of apo-R111M, especially the ligand entrance, is much less flexible than that of apo-CRABPII. Surprisingly, apo-R111M is more similar to holo-CRABPII than to apo-CRABPII in both structure and dynamical properties. The conformational and dynamical changes caused by the mutation are similar to those induced by binding of RA, although the magnitudes of the changes caused by the mutation are smaller than those induced by binding of RA. The results suggest that Arg111 plays a critical role in determining the structure and dynamical properties of CRABPII. PMID- 9737885 TI - Dissection of the sequence specificity of the holliday junction endonuclease CCE1 PMID- 9737884 TI - Mechanism of heparin activation of antithrombin: evidence for an induced-fit model of allosteric activation involving two interaction subsites. AB - The anticoagulant activation of the serpin antithrombin by heparin pentasaccharide DEFGH was previously shown to involve trisaccharide DEF first binding and inducing activation of the serpin, followed by disaccharide GH binding and stabilizing the activated state [Petitou et al. (1997) Glycobiology 7, 323-327; Desai et al. (1998) J. Biol. Chem. 273, 7478-7487]. In the present study, the role of conformational changes and charged residues of the GH disaccharide in the allosteric activation mechanism was investigated with variant pentasaccharides modified in the GH disaccharide. Perturbation of the conformational equilibrium of iduronate residue G through replacement of the nonessential 3-OH of this residue with -H resulted in parallel decreases in the fraction of residue G in the skew boat conformer (from 64 to 24%) and in the association constant for pentasaccharide binding to antithrombin [(2.6 +/- 0.3) fold], consistent with selective binding of the skew boat conformer to the serpin. Introduction of an additional sulfate group to the 3-OH of residue H flanking a putative charge cluster in the GH disaccharide greatly enhanced the affinity for the serpin by approximately 35-fold with only a small increase in the fraction of residue G in the skew boat conformation (from 64 to 85%). The salt dependence of binding, together with a recent X-ray structure of the antithrombin-pentasaccharide complex, suggested that the majority of the enhanced affinity of the latter pentasaccharide was due to direct electrostatic and hydrogen-bonding interactions of the H residue 3-O-sulfate with antithrombin. All variant pentasaccharides produced a normal enhancement of antithrombin fluoresence and normal acceleration of factor Xa inhibition by the serpin at saturating levels, indicating that conformational activation of antithrombin was not affected by the pentasaccharide modifications. Rapid kinetic studies were consistent with the altered affinities of the variant pentasaccharides resulting mostly from perturbed interactions of the reducing-end GH disaccharide with the activated antithrombin conformation and minimally to an altered binding of the nonreducing-end DEF trisaccharide to the native serpin conformation. Together, these results support a model in which the conformational flexibility of the G residue facilitates conversion to the skew boat conformer and thereby allows charged groups of the GH disaccharide to bind and stabilize the activated antithrombin conformation that is induced by the DEF trisaccharide. PMID- 9737886 TI - Antiinflammatory effects of euclyptol (1.8-cineole) in bronchial asthma: inhibition of arachidonic acid metabolism in human blood monocytes ex vivo. AB - Monoterpenes are prescribed to treat chronic obstructive airway disorders mainly because of their familiar secretolytic properties. The aim of this study was to investigate the effect of 1.8-cineole (Soledum) on arachidonic acid (AA) metabolism in blood monocytes of patients with bronchial asthma. Patients with bronchial asthma (n = 10) and healthy test subjects (n = 12) were included in the study. Production of the representative AA-metabolites LTB4 and PGE2 from isolated monocytes stimulated with the calcium ionophore A23187 were measured ex vivo before therapy with 1.8-cineole (3 x 200 mg/day), after three days of treatment (day 4) and four days after discontinuation of 1. 8-cineole (day 8). The production of LTB4 and PGE2 from monocytes ex vivo was significantly inhibited on day 4 in patients with bronchial asthma (-40.3%, n = 10 and -31.3%, p = 0.1, n = 3 respectively) as well as in healthy volunteers (-57.9%, n = 12 and -42.7%, n = 8 respectively). In conclusion, 1.8-cineole was shown to inhibit LTB4 and PGE2, both pathways of AA-metabolism. Further studies are needed to show that 1.8-cineole is suitable in the treatment of bronchial asthma. PMID- 9737887 TI - Follow-up in 103 patients with catecholamine-secreting tumours. AB - 103 patients from a group of 115 patients with catecholamine secreting tumours were reinvestigated 7.0 +/- 4.9 years following surgery. Throughout the follow-up period 15 patients had died. In four of them death was definitively, in seven subjects possibly associated to the primary endocrine disorder. Following surgery improvement of general well-being was documented in 85% of the patients. Hypertension was corrected in 61 %, but 26% of the patients remained hypertensive and symptoms of hypotension like orthostasis developed in 24%. A significant increase in weight (> 5 kg) was observed in 26% of the subjects throughout the follow-up period, but did not result in a higher prevalence of diabetes mellitus which had to be treated in 16% of the patients before and only 14% following surgery. However, palpitations, increased sweating and headache persisted in 16%, 17% and 12% of the patients, respectively. Symptoms of cardiac insufficiency developed in 32%. Persistent discomfort related to the scar was reported by 55% of the patients following lumbar surgery and by 30% of the subjects that were operated on via a transabdominal approach. Hence we conclude that surgery of catecholamine-secreting tumours results in an improvement of health and well being in most subjects according to objective criteria as well as to the judgement of the patients themselves. PMID- 9737888 TI - The splice-pattern of CD44 is altered in chronic pancreatitis exhibiting dysplastic changes. AB - Recent studies have shown that several splice variants of CD44, might be involved in tumor progression. Since chronic pancreatitis is suggested to be a risk factor for pancreatic cancer we investigated the splice pattern of CD44 in chronic pancreatitis to elucidate the role of CD44 in pancreas tumorigenesis. The expression of CD44-isoforms was examined in 40 specimens of chronic pancreatitis and 12 specimens of normal pancreas by immunohistochemistry, Westernblotting and exon specific RT-PCR. Pancreatic cancer tissue from two patients who developed pancreatic cancer 2 and 3 years following surgery for chronic pancreatitis were analyzed. Strong expression of CD44s was found in all cells, whereas the expression of CD44v6 was restricted to ductal cells. Westernblotting revealed an overexpression of CD44v6 in chronic pancreatitis as compared to normal pancreas. Exon specific analysis revealed an altered splice pattern of CD44, similar to that in pancreatic cancer, in 12.5% of the chronic pancreatitis specimens. Both patients who developed pancreatic cancer after chronic pancreatitis exhibited this altered splice pattern in both, chronic pancreatitis and pancreatic cancer. These results suggest that variant forms of CD44-mRNA might be expressed in early dysplastic alterations in chronic pancreatitis. PMID- 9737889 TI - Influence of clodronate on breast cancer cells in vitro. AB - One of the major therapies of bone metastasis is administration of clodronate. But the influence of clodronate alone on tumour cells is not quite clear. Radiotherapy and administration of clodronate increasingly are used in combination. The influence of clodronate on radiosensitivity of tumour cells is not known. METHODS: We used MDA-MB-435S and MCF-7 cells (breast cancer) in vitro and exposed the cells to clodronate in different concentrations and different short application times. In a second experiment we added graded doses of radiotherapy to the cell cultures. All experiments were done under standard conditions of the colony test. RESULTS: At different concentrations and different incubation times clodronate is able to reduce the cell survival of MDA-MB-435S cells, but not of MCF-7 cells. Even very low concentrations of clodronate in the cell culture medium are sufficient to reduce the tumour cell survival of MDA-MB 435S down to 59%. This reduction is time and concentration dependent. Using irradiation, clodronate has definitively no influence on the radiosensitivity of MDA-MB-435S cells in vitro, but the shoulder of the survival curve of MCF-7 cells is markedly reduced, demonstrating reduced repair of sublethal radiation damage. PMID- 9737891 TI - Resolution of severe Kaposi's sarcoma after initiation of antiretroviral triple therapy. AB - OBJECTIVES: To study the relationship between effective antiretroviral therapy, monitored by CD4 counts, and it s influence on the clinical course of AIDS associated Kaposi's sarcoma. METHODS: Four representative cases with AIDS and advanced Kaposi's sarcoma (KS) showed improvement of histologically proven KS s in various sites, including pulmonary disease, treated with liposomal doxorubicin. CD4 counts increased significantly during administration of triple antiretroviral therapy. In three cases chemotherapy cycles were extended and subsequently discontinued for 4, 14 and 4 months, respectively, without any relapse. In one other case interferonalpha therapy has been started overlapping with doxorubicin prior to permanent discontinuation of doxorubicin. CONCLUSIONS: Data of those patients suggest that in patients with increasing CD4 counts KS's chemotherapy intervals should be extended or even discontinued. In some patients change of therapy to interferon alpha can be considered. A potent combined antiretroviral therapy may enhance efficiency of KS treatment even in patients with high CD4 counts. PMID- 9737890 TI - In vitro study of human alveolar macrophages inflammatory mediator transcriptions and releases induced by soot FR 101, Printex 90, titandioxide and Chrysotile B. AB - Soot FR 101, Printex 90 and Chrysotile B are frequently found in indoor air pollutants phagocytized by alveolar macrophages (AM) involved in inflammatory pulmonary processes as, e.g. in cytokine secretions. The transcription factor NF kappaB has a role in the trans-duction pathway of proinflammatory cytokines like IL-1beta, IL-6 and TNF-alpha. We therefore investigated whether the transcription factor NF-kappaB and subsequent inflammatory cytokine secretions by AM are induced by exposure to these particles compared to the inert TiO2. AM were incubated for 90 min at particle concentrations of up to 100 microg/10(6) cells. Sequential reverse transcription and semiquantitative cDNA amplification (RT-PCR) was used to measure NF-kappaB and cytokine mRNA expressions. Compared to control exposures these particles induced an up to 4.6-fold increase in gene expression of the transcription factor NF-kappaB (p < 0.01), resulting in up to 12.9-fold enhanced transcription rates of IL-1beta, IL-6 and TNF-alpha (p <0.05). The particles and fibre dependent increases in mRNA reached maximum levels at 90 min post exposure. After an exposure time of 8 hrs, IL-1beta, IL-6 and TNF-alpha proteins, measured by enzyme-linked immunosorbent assays (ELISA), were significant elevated in supernatants of AM, revealing an up to 30.5-fold increase in TNF-alpha secretion rates (p <0.01). Our results suggest that exposure of human AM to soot FR 101, Printex 90, TiO2 and Chrysotile B induce the transcription and production of proinflammatory cytokines via NF-kappaB and may play an important role in the pathogenesis of airway disease and lung parenchymal injury. PMID- 9737892 TI - Gene therapy and medical genetics on Internet. AB - In this report we consider the development of the Internet, from its origins as a military invention in the times of the cold war to its present day role, together with the World Wide Web, as a means of global communication which plays a key role in medical research and particularly in medical genetics. A few of the major genetics related research projects and gene research centers are introduced and their aims are briefly discussed. Detailed information about chromosome and gene mapping, together with sequence and structure databases, can be easily and rapidly accessed through the Internet. A variety of web-sites are briefly described and then listed at the end of the report, which will serve as a useful starting point from which the interested reader can access an almost endless source of genetics related information on the Internet. Finally, some of the ethical, legal and social implications of the links between gene therapy and the Intemet are considered. PMID- 9737893 TI - Treatment of AIDS-related complications. 12th World-AIDS-Conference, June 1998, Geneva. PMID- 9737894 TI - A buried polar interaction imparts structural uniqueness in a designed heterodimeric coiled coil PMID- 9737895 TI - Protein farnesyltransferase: structure and implications for substrate binding PMID- 9737896 TI - Quality participation. PMID- 9737897 TI - Steering toward quality. PMID- 9737898 TI - Quality of care for hip fracture patients. PMID- 9737899 TI - Using the EU to improve quality. PMID- 9737900 TI - Managing information. PMID- 9737901 TI - Quality in concert. PMID- 9737902 TI - Euroquan. Quality across frontiers. PMID- 9737903 TI - An accessory fissure in the lower lobe of the right lung. AB - We report here an accessory fissure in the right lung. Accessory fissures are described as clefts of various depths lined by two layers of visceral pleura. Anomalies of location of the lungs may be produced by the accessory fissures. In our case, an unusual accessory fissure in the right lung was found in a 50 years old male cadaver. The accessory fissure was located between the superior and basal segments of the lower lobe of the right lung. Its depth was around 3 cm as an average value. Its length was found to be 12 cm on the surface of the lung. In conclusion, a surgeon must always remember the anatomical variations of the location of the lungs especially in lobectomies and in segmental resection. From the radiological aspect, an accessory fissure is important in that it can be mistaken for lung pathologies. PMID- 9737904 TI - [Degeneration and regeneration of striated skeletal muscle fibers in Duchenne muscular dystrophy]. AB - Like all other muscular dystrophies, Duchenne muscular dystrophy is characterized by the coexistence of degenerative lesions of the muscle fibers and of regenerative changes. The present study has been carried out in order to precise the degree of regeneration at different stages of the disease, by analyzing the expression of several markers of cell proliferation and of muscular differentiation. In the two affected foetuses of our series, the m. quadriceps is histologically normal, except for the absent expression of immunoreactive dystrophin. The quadriceps from the eight children of our series (20 months-16 years) all present clear dystrophic changes. Muscle regeneration is characterized by activation of the satellite cells, by their multiplication followed by their fusion giving birth to regenerative fibers. By studying the expression of muscular markers (vimentin, desmin, isoforms of the myosin heavy chains), it has been possible to define more precisely the degree of maturation and of differentiation of these regenerative fibers. Our results suggest that an abortive regeneration of the muscle fibers in Duchenne muscular dystrophy can explain, at least partly, the progressive evolution of this disease. PMID- 9737905 TI - Bilateral anomaly of the semispinalis capitis muscle. AB - Bilateral variation of the semispinalis capitis muscle in a 48 year old male cadaver was encountered during routine dissectionin our department. A review of the literature reveals no additional reports of similar anomalies. PMID- 9737906 TI - An anatomic variation of the stylomastoid foramen. AB - During the examination of cranial bones, we observed an abnormality in one of the right temporal bones. In this variation, the stylomastoid foramen was not completely formed on the temporal bone and instead of a foramen, there was a sulcus in its localization on the bone. The articulation with the occipital bone transformed this sulcus into a foramen. This variation was not reported in the literature. PMID- 9737907 TI - A scapula with an unreported canal. AB - During the practical lectures in our department, we encountered an abnormality on the left scapula of an adult human. In this case, an elongated canal was found just posterior and inferior to the coracoid process and the scapular notch was not observed on this bone. Knowing about the anatomic variations of this region may have an importance for the surgeons. PMID- 9737908 TI - Microanatomy of the vasa vasorum of the human thoracic aorta: a study utilizing a polyester resin casting technique. AB - We have studied the vasa vasorum of the human thoracic aorta using a polyester resin casting technique, coupled with the tissue clearing method of Spalteholz. We found that the vasa vasorum originated from intercostal arteries. They ramified and formed networks within the aortic adventitia and the outer layers of the media. Venous networks were identified in the adventitia as wall. In addition to being a usefull research tool, the methodology utilized in this study produces specimens that can be used in teaching vascular anatomy to medical students. PMID- 9737909 TI - [Utilization of thermosettable compounds in anatomic research]. AB - The authors described a method of vascular injection with a coloured silicon rubber. The injected material was a biocomponent silicon elastomer, with ambiant temperature room vulcanizing. It was supple, easily dissequable and diffuse well into all small caliber vessels. The soft pressure injection did not cause neither material collection by vessels rupture nor anatomic structure distortion. This material could constitute an excellent alternative to coloured latex injection. PMID- 9737910 TI - [Anatomic study via dissection of the common fibular nerve: etiopathogenic consideration]. AB - In order to understand and prevent the particular rate of the common fibular nerve palsy during the traumatisms of the main stem of the sciatic nerve; the authors have studied through dissection the osteomuscular and aponeurotic relationships of 38 common fibular nerves. They blame it on the relations between the nerve and the proximal ends of the fibula; the nerve can be stretched on and mainly on its fixation on the crural fascia by fibrous tracts seen in 78.94 of the cases. PMID- 9737911 TI - Unusual bifurcation-level variation of the left common carotid artery: anatomic presentation of a surgical case. AB - We report an anatomical variation of the left common carotid artery in a 65 year old patient with larynx carcinoma, that was detected during a radical neck dissection. The bifurcation of the artery was at the level of sixth cervical vertebra that is significantly lower than the commonly accepted normal level. To the best of our knowledge, there is no other report of a similar variation in the medical literature. PMID- 9737912 TI - [Human cervico-facial morphogenesis. Evaluation of acquired data and current outlook. (Part 1: facial morphogenesis)]. AB - The embryonic development of the face has been studied in many reviews, this work purposes only to clear up some points which remain obscure concerning cervico facial morphogenesis. In the first part of this study only the facial development, properly speaking, is considered, although it cannot be separate of cervical development to which a second study will be reserved. In the present study we recall the particular aspects of the neurulation in the cephalic area, then the establishment of the facial processes. Then we approach among other things the way to consider the maxillary process with regard to the other facial processes. After is considered constitution and natured of the prechordal plate which has been diversely explained. Finally, the modelling of the face is evocated, in which the dissociation between the olfactive and buccal spheres is pointed out, with the disparition of the muzzle, as it is established in the haplorhinae, a class of primates in which the human being is involved. This phenomenon raises different questions, in particular about the relation of this disposition with the nasoseptal center, the medial part of the nasodorsal center. PMID- 9737913 TI - Arterial branches to the Palmaris Longus muscle. AB - The use of the long palmar muscle in surgeries of muscular transposition and transplantation in orthopedic and plastic surgeries has encouraged the investigation of arterial branches for the long palmar muscle. This study was carried out in 58 upper limbs of child cadavers aged up to one year. It was observed that the muscle was not present in 31.04% of the forearms. It more frequently presents one to two arterial branches which arise mainly, from the ulnar artery (85.71%) and less frequently from brachial artery (22.86%). The arterial branches penetrate the muscle through the posterior face, 62.71% being at the proximal third and 33.90% at the medium third. The most frequent models of the branch disposition were from one to two branches of the ulnar artery which penetrate the proximal thirds (28.57%) and two branches of the ulnar artery, the median and proximal thirds, respectively (17.14%). PMID- 9737914 TI - Study on optic microscope of the muscle and elastic components of the aorto-iliac bifurcation septum and initial segment of the middle sacral artery. AB - In this work the aorto-iliac bifurcation septum and the initial segment of the middle sacral artery were analyzed through optic microscope. 20 anatomical pieces of corpses from subjects of both sexes aging from 20 to 33 years were examined. After collection of pieces and fixation in formaline for 48 hours, histological sections of 20 microns were made and stained with the methods of Weigert, Verhoeff and Picrofucsine of Van Gieson. It was observed that the middle sacral artery arises from the posterior aortic wall laterally to the aorto-iliac bifurcation septum. The sequential analysis of the sections showed that the septum is lined by a thickening of connective tissue along its length and elastic fibers predominate on its central portion. Bundles of muscle fibers prevail on the lateral and posterior regions. It was observed, above the origin of the middle sacral artery, a reorganisation of tissues which represents, more caudally, the ostium of the artery. It was verified that the middle sacral artery arises at the aorto-iliac bifurcation septum, which is a transitional region and should be classified of mixed type and called aorto-septo-sacral transition. PMID- 9737915 TI - [Branchial cyst of unusual localization: report of a case and considerations on organogenesis]. AB - We report a case of branchial cyst of unusual location. A asymptomatic 41-year old man had a nontender deeply located left neck mass. Sonography, CT scan, and MRI showed a cystic lesion posterior to the sternocleidomastoid muscle. The diagnosis of branchial cyst much debated because of this atypical location was confirmed by histologic analysis after surgical resection. During organogenesis, the important caudal proliferation of the second branchial arch generates a transient cavity, the cervical sinus, which finally becomes obliterated. The incomplete obliteration of which can result in a sinus, fistula or cyst. Such cysts typically lie at the level of the mandibular angle, anterior to the sternocleidomastoid muscle. This location has been regarded as a major diagnosis criteria, but it is not absolute. The sternocleidomastoid muscle develops apart from the branchial apparatus, caudally and anteriorly. As a result the cysts which are located on an inferior portion of the cervical sinus can lie posterior to this muscle. PMID- 9737916 TI - An examination of the relationship between foot length, T1, T2, T3, T4, T5 (toe lengths), ankle circumference and calf circumference of Turkish University students aged between 17 and 25. AB - In this study, the relationship, between Foot Length, Toe Lengths, Ankle Circumference and Calf Circumference of the students aged 17-25 was examined. When looked at the correlation coefficients, a significant relationship between Foot Length, Ankle Circumference, Calf Circumference and T1, T2, T3, T4, T5 (Toe Lengths) was discovered (p < 0.05). However, from the statistical point of view the relationship between only Foot Length and Ankle Circumference in the Males was discovered to be bigger than the Females (p < 0.05). Whereas, the other measurement in both Males and Females were relatively close to each other. During the study it was discovered that while the relationship between Ankle Circumference and Calf Circumference in both sexes was statistically significant (p < 0.05), the relationship between Ankle Circumference and T1, T2, T3, T4 was also statistically significant, but this relationship in Males were bigger than that of Females (p < 0.05). When the regression analyse was carried out, it was observed that there was a linear relationship between Foot Length and T1-T5. This indicated that Turkish people has bigger Ankle & Calf Circumferences and Foot Length than Japanese people. PMID- 9737917 TI - [Kinematics of the pelvic girdle and the thoracic and lumbar segments in the course of the lateral inclinations and rotations of the spine]. AB - Physiological displacements of the trunk are the addition of these of the pelvic girdle and thoracic and lumbar spine segments. For a long time (3), this conjunction had been noticed but without numbered precisions. The purpose of this communication is to appreciate quantitatively, from a series of 16 subjects, the respective share of each components during lateral bending movements in the frontal plan and movements of rotation in the transverse plan. It results from this work that the pelvic girdle presents as reduced amplitude (4 degrees) in lateral bending on the other hand, displacement predominate to the level of thoracic spine (50 degrees). In rotation pelvic displacements are very important (30 degrees), while the spine so thoracic that lumbar has a weak participation (inferior to 5 degrees). Moderated abduction of hips increases by significant manner the motility of the pelvic girdle. PMID- 9737918 TI - CAP, the -45 region, and RNA polymerase: three partners in transcription initiation at lacP1 in Escherichia coli. AB - The lac operon of Escherichia coli is positively regulated by the catabolite activator protein (CAP) bound upstream of the -45 region (CAP binding is centered at -61.5; the -45 region extends from -50 to -38). Certain mutations within the 45 region generate sequences that resemble UP elements in base composition and mimic the stimulation by the rrnBP1 UP element, yielding up to 15-fold stimulation in vivo. These -45 region "UP mutants" are compromised in their CAP stimulation. CAP and UP elements do not act in a fully additive manner in vivo at the lac operon. Transcription assays with the wild-type lac promoter and an UP mutant of lac indicate that CAP and UP DNA also fail to act in a completely additive manner in vitro. RNA polymerase can stabilize CAP binding to promoter DNA with a -45 region UP element against a heparin challenge. This shows that CAP and the UP DNA do not compete for the alpha-CTD as a mechanism for their lack of additivity. CAP and UP elements both demonstrate decreased stimulation of transcription as RNA polymerase concentration is increased from 0.05 to 10 nM in in vitro transcription experiments. In addition CAP also stimulates transcription in a manner that does not decrease as RNA polymerase is varied over this concentration range. This invariable stimulation is by two- to threefold and occurs both in vivo and in vitro. It is not dependent upon the alpha-CTD of RNA polymerase and is maintained in the presence of the AR1 CAP mutant HL159. This two- to threefold invariable CAP stimulation appears to depend on the -45 region sequence as our -45 region mutants demonstrate different responses to HL159 CAP stimulation in vivo. PMID- 9737919 TI - Group II intron mobility in yeast mitochondria: target DNA-primed reverse transcription activity of aI1 and reverse splicing into DNA transposition sites in vitro. AB - The retrohoming of the yeast mtDNA intron aI1 occurs by a target DNA-primed reverse transcription (TPRT) mechanism in which the intron RNA reverse splices directly into the recipient DNA and is then copied by the intron-encoded reverse transcriptase. Here, we carried out biochemical characterization of the intron encoded reverse transcriptase and site-specific DNA endonuclease activities required for this process. We show that the aI1 reverse transcriptase has high TPRT activity in the presence of appropriate DNA target sites, but differs from the closely related reverse transcriptase encoded by the yeast aI2 intron in being unable to use artificial substrates efficiently. Characterization of TPRT products shows that the fully reverse spliced intron RNA is an efficient template for cDNA synthesis, while reverse transcription of partially reverse spliced intron RNA is impeded by the branch point. Novel features of the aI1 reaction include a prominent open-circular product in which cDNAs are incorporated at a nick at the antisense-strand cleavage site. The aI1 endonuclease activity, which catalyzes the DNA cleavage and reverse splicing reactions, is associated with ribonucleoprotein particles containing the intron-encoded protein and the excised intron RNA. As shown for the aI2 endonuclease, both the RNA and protein components are used for DNA target site recognition, but the aI1 protein has less stringent nucleotide sequence requirements for the reverse splicing reaction. Finally, perhaps reflecting this relaxed target specificity, in vitro experiments show that aI1 can reverse splice directly into ectopic mtDNA transposition sites, consistent with the previously suggested possibility that this mechanism is used for ectopic transposition of group II introns in vivo. PMID- 9737920 TI - Mutant alleles of the MRS2 gene of yeast nuclear DNA suppress mutations in the catalytic core of a mitochondrial group II intron. AB - Previous studies show that some yeast strains carrying point mutations of domain 5 that block splicing of a mitochondrial group II intron yield spontaneous revertants in which splicing is partially restored by dominant mutations of nuclear genes. Here we cloned and sequenced the suppressor allele of one such gene, and found it to be a missense mutation of the MRS2 gene (MRS2-L232F). The MRS2 gene was first implicated in group II intron splicing by the finding that overexpression of the wild-type gene weakly suppresses the splicing defect of a mutation of another intron. Tetrad analysis showed that independently isolated suppressors of two other domain 5 mutations are also allelles of the MRS2 gene and DNA sequencing identified a new missense mutation in each strain (MRS2-T230I and MRS2-L213M). All three suppressor mutations cause a temperature-sensitive respiration defect that is dominant negative in heterozygous diploids, but those strains splice the mutant intron at the elevated temperature. The three mutations are in a domain of the protein that is likely to be a helix-turn-helix region, so that effects of the mutations on protein-protein interactions may contribute to these phenotypes. These mutations suppress the splicing defect of many, but not all, of the available splicing defective mutations of aI5gamma, including mutations of several intron domains. Protein and RNA blot experiments show that the level of the protein encoded by the MRS2 gene, but not the mRNA, is elevated by these mutations. Interestingly, overexpression of the wild-type protein restores much lower levels of splicing than were obtained with similar elevated levels of the mutated Mrs2 proteins. The splicing phenotypes of these strains suggest a direct role for Mrs2 protein on group II intron splicing, but an indirect effect is not yet ruled out. PMID- 9737921 TI - Genome plasticity in the distal tail fiber locus of the T-even bacteriophage: recombination between conserved motifs swaps adhesin specificity. AB - The adsorption specificity of the T-even phages is determined by the protein sequence near the tip of the long tail fibers. These adhesin sequences are highly variable in both their sequence and specificity for bacterial receptors. The tail fiber adhesin domains are located in different genes in closely related phages of the T-even type. In phage T4, the adhesin sequence is encoded by the C-terminal domain of the large tail fiber gene (gene 37), but in T2, the adhesin is a separate gene product (gene 38) that binds to the tip of T2 tail fibers. Analysis of phage T6 and Ac3 sequences reveals additional variant forms of this locus. The tail fiber host specificity determinants can be exchanged, although the different loci have only limited homology. Chimeric fibers can be created by crossovers either between small homologies within the structural part of the fiber gene or in conserved motifs of the adhesin domain. For example, the T2 adhesin determinants are flanked by G-rich DNA motifs and exchanges involving these sequences can replace the specificity determinants. These features of the distal tail fiber loci genetically link their different forms and can mediate acquisition of diverse host range determinants, including those that allow it to cross species boundaries and infect taxonomically distant hosts. PMID- 9737922 TI - Antibiotic inhibition of RNA catalysis: neomycin B binds to the catalytic core of the td group I intron displacing essential metal ions. AB - The aminoglycoside antibiotic neomycin B induces misreading of the genetic code during translation and inhibits several ribozymes. The self-splicing group I intron derived from the T4 phage thymidylate synthase (td) gene is one of these. Here we report how neomycin B binds to the intron RNA inhibiting splicing in vitro. Footprinting experiments identified two major regions of protection by neomycin B: one in the internal loop between the stems P4 and P5 and the other in the catalytic core close to the G-binding site. Mutational analyses defined the latter as the inhibitory site. Splicing inhibition is strongly dependent on pH and Mg2+ concentration, suggesting electrostatic interactions and competition with divalent metal ions. Fe2+-induced hydroxyl radical (Fe-OH.) cleavage of the RNA backbone was used to monitor neomycin-mediated changes in the proximity of the metal ions. Neomycin B protected several positions in the catalytic core from Fe-OH. cleavage, suggesting that metal ions are displaced in the presence of the antibiotic. Mutation of the bulged nucleotide in the P7 stem, a position which is strongly protected by neomycin B from Fe-OH. cleavage and which has been proposed to be involved in binding an essential metal ion, renders splicing resistant to neomycin. These results allowed the docking of neomycin to the core of the group I intron in the 3D model. PMID- 9737923 TI - Cointegrase, a naturally occurring, truncated form of IS21 transposase, catalyzes replicon fusion rather than simple insertion of IS21. AB - The bacterial insertion sequence IS21 contains two genes, istA and istB, which are organized as an operon. IS21 spontaneously forms tandem repeats designated (IS21)2. Plasmids carrying (IS21)2 react efficiently with other replicons, producing cointegrates via a cut-and-paste mechanism. Here we show that transposition of a single IS21 element (simple insertion) and cointegrate formation involving (IS21)2 result from two distinct non-replicative pathways, which are essentially due to two differentiated IstA proteins, transposase and cointegrase. In Escherichia coli, transposase was characterized as the full length, 46 kDa product of the istA gene, whereas the 45 kDa cointegrase was expressed, in-frame, from a natural internal translation start of istA. The istB gene, which could be experimentally disconnected from istA, provided a helper protein that strongly stimulated the transposase and cointegrase-driven reactions. Site-directed mutagenesis was used to express either cointegrase or transposase from the istA gene. Cointegrase promoted replicon fusion at high frequencies by acting on IS21 ends which were linked by 2, 3, or 4 bp junction sequences in (IS21)2. By contrast, cointegrase poorly catalyzed simple insertion of IS21 elements. Transposase had intermediate, uniform activity in both pathways. The ability of transposase to synapse two widely spaced IS21 ends may reside in the eight N-terminal amino acid residues which are absent from cointegrase. Given the 2 or 3 bp spacing in naturally occurring IS21 tandems and the specialization of cointegrase, the fulminant spread of IS21 via cointegration can now be understood. PMID- 9737924 TI - Shared receptor components but distinct complexes for alpha and beta interferons. AB - The type I interferon family includes 13 alpha, one omega and one beta subtypes recognized by a complex containing the receptor subunits ifnar1 and ifnar2 and their associated Janus tyrosine kinases, Tyk2 and Jak1. To investigate the reported differences in the way that alpha and beta interferons signal through the receptor, we introduced alanine-substitutions in the ifnar2 extracellular domain, and expressed the mutants in U5A cells, lacking endogenous ifnar2. A selection, designed to recover mutants that responded preferentially to alpha or beta interferon yielded three groups: I, neutral; II, sensitive to alpha interferon, partially resistant to beta interferon; III, resistant to alpha interferon, partially sensitive to beta interferon. A mutant clone, TMK, fully resistant to alpha interferon with good sensitivity to beta interferon, was characterized in detail and compared with U5A cells complemented with wild-type ifnar2 and also with Tyk2-deficient 11.1 cells, which exhibit a similar alpha unresponsive phenotype with a partial beta interferon response. Using anti receptor antibodies and mutant forms of beta interferon, three distinct modes of ligand interaction could be discerned: (i) alpha interferon with ifnar1 and ifnar2; (ii) beta interferon with ifnar1 and ifnar2; (iii) beta interferon with ifnar2 alone. We conclude that alpha and beta interferons signal differently through their receptors because the two ligand subtypes interact with the receptor subunits ifnar 1 and ifnar2 in entirely different ways. PMID- 9737925 TI - Assembly and architecture of invertebrate cytoplasmic intermediate filaments reconcile features of vertebrate cytoplasmic and nuclear lamin-type intermediate filaments. AB - The two major intermediate filament (IF) proteins from the esophagus epithelium of the snail Helix pomatia and the two major IF proteins from muscle tissue of the nematode Ascaris suum were investigated under a variety of assembly conditions. The lowest-order complexes from each of the four protostomic invertebrate (p-INV) IF proteins are parallel, unstaggered dimers involving two stranded alpha-helical coiled coil formation of their approximately 350 amino acid residue central rod domain (i.e. long-rod). In the electron microscope these are readily recognized by their distinct approximately 56 nm long rod with two globular domains (i.e. representing the non-helical carboxy-terminal tail domain of the p-INV IF proteins) attached at one end, closely resembling vertebrate lamin dimers. The next-higher-order oligomers are tetramers, which are easily recognized by their two pairs of globular tail domains attached at either end of a approximately 72 nm long central rod portion. According to their size and shape, these tetramers are built from two dimers associated laterally in an antiparallel, approximately half-staggered fashion via the amino-terminal halves of their rod domains. This is similar to the NN-type tetramers found as the most abundant oligomer species in all types of vertebrate cytoplasmic IF proteins, which contain a approximately 310 amino acid residue central rod domain (i.e. short-rod). As a first step toward filament formation, the p-INV IF tetramers anneal longitudinally into protofilaments by antiparallel CC-type association of the carboxy-terminal halves of their dimer rods. The next step involves radial growth, occurring initially through lateral association of two four-chain protofilaments into octameric subfibrils, which then further associate into mature, full-width filaments. Head-to-tail polymers of dimers and paracrystalline fibers commonly observed with vertebrate lamins were only rarely seen with p-INV IF proteins. The globular domains residing at the carboxy-terminal end of p-INV IF dimers were studding the surface of the filaments at regular, approximately 24.5 nm intervals, thereby giving them a "beaded" appearance with an axial periodicity of about 24.5 nm, which is approximately 3 nm longer than the corresponding approximately 21.5 nm repeat pattern exhibited by short-rod vertebrate IFs. PMID- 9737926 TI - Solution structure of a Na cation stabilized DNA quadruplex containing G.G.G.G and G.C.G.C tetrads formed by G-G-G-C repeats observed in adeno-associated viral DNA. AB - We have applied NMR and molecular dynamics computations including intensity based refinement to define the structure of the d(G-G-G-C-T4-G-G-G-C) dodecanucleotide in 100 mM NaCl solution. The G-G-G-C sequence is of interest since it has been found as tandem repeats in the DNA sequence of human chromosome 19. The same G-G G-C sequence is also seen as islands in adeno-associated virus, a human parvovirus, which is unique amongst eukaryotic DNA viruses in its ability to integrate site-specifically into a defined region of human chromosome 19. The d(G G-G-C-T4-G-G-G-C) sequence forms a quadruplex in Na cation containing solution through head-to-tail dimerization of two symmetry-related stem-hairpin loops with adjacent strands antiparallel to each other around the quadruplex. The connecting T4 loops are of the lateral type, resulting in a quadruplex structure containing two internal G.G.G.G tetrads flanked by G.C.G.C tetrads. The G(anti).G(syn).G(anti).G(syn) tetrads are formed through dimerization associated hydrogen bonding alignments of a pair of Hoogsteen G(anti).G(syn) mismatch pairs, while the G(anti).C(anti).G(anti).C(anti) tetrads are formed through dimerization associated bifurcated hydrogen bonding alignments involving the major groove edges of a pair of Watson-Crick G.C base-pairs. The quadruplex contains two distinct narrow and two symmetric wide grooves with extensive stacking between adjacent tetrad planes. The structure of the quadruplex contains internal cavities that can potentially accommodate Na cations positioned between adjacent tetrad planes. Three such Na cations have been modeled into the structure of the d(G-G-G-C-T4-G-G-G-C) quadruplex. Finally, we speculate on the potential role of quadruplex formation involving G.G.G.G and G.C.G.C tetrads during the integration of the adeno-associated parvovirus into its target on human chromosome 19, both of which involve stretches of G-G-G-C sequence elements. PMID- 9737927 TI - A K cation-induced conformational switch within a loop spanning segment of a DNA quadruplex containing G-G-G-C repeats. AB - We have identified a unique structural transition (in slow exchange on the NMR time scale) in the tertiary fold of the d(G-G-G-C-T4-G-G-G-C) quadruplex on proceeding from Na+ to K+ as counterion in aqueous solution. Both monovalent cation-dependent conformations exhibit certain common structural features, which include head-to-tail dimerization of two symmetry-related stem-hairpin loops, adjacent strands which are antiparallel to each other and adjacent stacked G(syn).G(anti). G(syn).G(anti) tetrads in the central core of the quadruplexes. The Na and K cation stabilized structures of the d(G-G-G-C-T4-G-G-G-C) quadruplexes differ in the conformations of the T-T-T-T loops, the relative alignment of G.C base-pairs positioned opposite each other through their major groove edges and potentially in the number of monovalent cation binding sites. We have identified potential K cation binding cavities within the symmetry-related T T-T-G segments, suggesting the potential for two additional monovalent cation binding sites in the K cation-stabilized quadruplex relative to its Na cation stabilized counterpart. Modeling studies suggest that the major groove edges of guanine residues in Watson-Crick G.C base-pairs could potentially be bridged by coordinated K cations in the d(G-G-G-C-T4-G-G-G-C) quadruplex in KCl solution in contrast to formation of G.C.G.C tetrads for the corresponding quadruplex in NaCl solution. Our results defining the molecular basis of a Na to K cation-dependent conformational switch in the loop spanning segment of the d(G-G-G-C-T4-G-G-G-C) quadruplex may have relevance to recent observations that specific K cation coordinated loop conformations within quadruplexes exhibit inhibitory activity against HIV integrase. PMID- 9737928 TI - Structural characterisation of apoflavodoxin shows that the location of the stable nucleus differs among proteins with a flavodoxin-like topology. AB - The structural characteristics of Azotobacter vinelandii apoflavodoxin II have been determined using multidimensional NMR spectroscopy. Apoflavodoxin has a stable, well-ordered core but its flavin binding region is flexible. The local stability of apoflavodoxin was probed using hydrogen/deuterium exchange measurements. The existence of an apoflavodoxin equilibrium folding intermediate is inferred from the non-coincidence of CD and fluorescence unfolding curves obtained for the guanidinium hydrochloride induced unfolding of apoflavodoxin. We suggest that the structured part of the putative intermediate is composed of the elements of secondary structure which have the slowest exchanging amide protons in the native protein. These elements are strands beta1, beta3, beta4 and beta5a and helices alpha4 and alpha5. We propose that it is a general feature of flavodoxins that the stable nucleus resides in the C-terminal part of these proteins. The results on flavodoxin are compared with those on two sequentially unrelated proteins sharing the flavodoxin-like fold: Che Y and cutinase. It is shown that the stable nucleus is found in different parts of the flavodoxin-like topology. PMID- 9737929 TI - A 2.8 A resolution structure of 6-phosphogluconate dehydrogenase from the protozoan parasite Trypanosoma brucei: comparison with the sheep enzyme accounts for differences in activity with coenzyme and substrate analogues. AB - The three-dimensional structure of 6-phosphogluconate dehydrogenase (6PGDH) from the parasitic protozoan Trypanosoma brucei has been solved at 2.8 A resolution. This pentose phosphate pathway enzyme is NADP-dependent; NADPH generated in the reaction protects against oxidative stress. The enzyme crystallises in the space group P3121 with a dimer in the asymmetric unit and cell dimensions a=b=135.13 A, c=116.74 A, alpha=beta=90 degrees, gamma=120 degrees. The structure has refined to R=18.6% (Rfree=27.3%) with good geometry. The amino acid sequence of T. brucei 6PGDH is only 35% identical to that of the sheep liver enzyme and significant activity differences have been observed. The active dimer assembles with the C terminal tail of one subunit threaded through the other, forming part of the substrate binding site. The tail of T. brucei 6PGDH is shorter than that of the sheep enzyme and its terminal residues associate tightly with the second monomer. The three-dimensional structure shows this generates additional interactions between the subunits close to the active site; the coenzyme binding domain is thereby associated more tightly with the helical domain. Three residues, conserved in all other known sequences, are important in creating a salt bridge between monomers close to the substrate binding site. The differences could explain the 200-fold enhanced affinity observed for the substrate analogue 6 phospho-2-deoxy-D-gluconate and suggest targets for anti-parasite drug design. The coenzyme binding domain of 6PGDH has a beta-alpha-beta fold; while in most species the "fingerprint" sequence is GxAxxG, in the T. brucei enzyme it is GxGxxG. Additional interactions between the enzyme and the coenzyme bis-phosphate are likely in the parasite 6PGDH, accounting for greater inhibition (40-fold) of 2'5'-ADP. While the core of the T. brucei dimer was restrained during refinement, several conformational differences have been found between the monomers; those at the coenzyme binding site suggest the molecule could be asymmetric during the enzyme reaction. PMID- 9737930 TI - Differential nucleosome positioning on Xenopus oocyte and somatic 5 S RNA genes determines both TFIIIA and H1 binding: a mechanism for selective H1 repression. AB - In Xenopus somatic cells histone H1 effects the transcriptional repression of oocyte type 5 S RNA genes, without altering the transcription of the somatic type 5 S RNA genes. Using an unambiguous nucleosome mapping method we find substantial differences between the multiple in vitro nucleosome positions on the two types of genes. These nucleosome positions determine both transcription factor and H1 binding, allowing TFIIIA to bind more efficiently to nucleosomes containing the somatic 5 S RNA gene than to nucleosomes on the oocyte 5 S RNA gene. Significantly, in a binding competition between TFIIIA and H1, TFIIIA preferentially binds to the somatic nucleosome whereas H1 preferentially binds to the oocyte nucleosome, excluding TFIIIA binding. These results strongly suggest that nucleosome positioning plays a key role in the regulation of transcription of 5 S RNA genes and provide a molecular mechanism for the selective repression of the oocyte 5 S RNA genes by H1. PMID- 9737931 TI - Corrigendum PMID- 9737933 TI - Frequency-dependence of myocardial energetics in failing human myocardium as quantified by a new method for the measurement of oxygen consumption in muscle strip preparations. AB - Diastolic dysfunction at high heart rates may be associated with increased myocardial energy consumption. Frequency-dependent changes of isometric force and oxygen consumption (MVO2) were investigated in strip preparations from endstage failing human hearts exhibiting various degrees of diastolic dysfunction. MVO2 was determined by a new method which was validated. When stimulation rate was increased from 40 to 200 min-1 (n=7), developed force decreased from 16.5+/-4.3 to 7.9+/-2.9 mN/mm2 (P<0.01), diastolic force increased from 15.9+/-3.2 to 22.0+/ 3.0 mN/mm2 (P<0.01), and total MVO2 increased from 2.6+/-0.6 to 4.7+/-0.9 ml/min/100 g (P<0.025). Resting MVO2 and resting force were 1.8+/-0.4 ml/min/100 g and 15.9+/-3.0 mN/mm2, respectively. After addition of 30 mm 2,3-butanedione monoxime (BDM) to inhibit crossbridges, resting MVO2 and resting force decreased by 46% (P<0.05) and 15% (P<0.01), respectively, indicating the presence of active force generation in unstimulated failing human myocardium. In each muscle preparation, there was a significant correlation between force-time integral (FTI) and total MVO2 (r=0.96+/-0.01). The strength of these correlations did not vary with the contribution of diastolic FTI to total FTI. The ratio of activity related MVO2 to developed FTI, an inverse index of the economy of contraction, increased depending on the rise of diastolic FTI at higher stimulation rates. In conclusion, in failing human myocardium, diastolic force development is occurring at the same energy expenditure as systolic force generation. Therefore, in muscle preparations with disturbed diastolic function economy of contraction decreases with higher stimulation rates, depending on the rise of diastolic force. PMID- 9737934 TI - Angiotensin II modulates ANP-R2/ANP-C receptor-mediated inhibition of adenylyl cyclase in vascular smooth muscle cells: role of protein kinase C. AB - In the present studies, we have investigated the modulation of atrial natriuretic peptide (ANP) receptor of R2 subtype (ANP-R2/ANP-C) coupled to adenylyl cyclase/cAMP signal transduction system by angiotensin II (angII). C-ANF4-23 [des(Gln18, Ser19, Gln20, Leu21, Gly22)ANF4-23-NH2] and AngII inhibited adenylyl cyclase activity in a concentration-dependent manner in vascular smooth muscle cells (VSmc A-10). The maximal inhibitions observed were about 40 and 30%, respectively, with an apparent Ki of about 1 and 10 nm. Pretreatment of the cells with AngII resulted in the attenuation of both C-ANF4-23 and AngII-mediated inhibitions of adenylyl cyclase, without altering [125I]-ANF binding. The levels of Gialpha-2 and Gialpha-3 proteins as determined by immunoblotting were also augmented by AngII treatment. In addition, AngII treatment stimulated the phosphorylation of Gialpha2 but not of Gialpha3 or ANP-C receptor, as revealed by immunoprecipitation of the proteins using specific antibodies after prelabelling the cells with [32P]orthophosphate. Staurosporine and chelerythrine, protein kinase C (PKC) inhibitors at 1 and 100 nm, respectively, prevented the AngII mediated desensitization of C-ANF 4-23-sensitive adenylyl cyclase. In addition, the AngII-mediated phosphorylation of Gialpha2 protein was also inhibited partially by about 35% by staurosporine treatment. These results suggest that the attenuation of C-ANF4-23-mediated inhibition of adenylyl cyclase activity by AngII may not be attributed to the downregulation of receptors or to the decreased levels of G-proteins, and may involve PKC-dependent mechanisms. PMID- 9737935 TI - The effect of nicotine on DNA repair in adult myocytes. AB - Recent findings have demonstrated that terminally differentiated adult ventricular myocytes are capable of repairing DNA that has been damaged by exposure to oxygen free radicals. Despite the potential importance of DNA repair in cells that may survive many decades after injury, little is known about the mechanisms or regulation of repair. Since tobacco use has a well-defined role in the epidemiology and pathophysiology of heart disease, we tested the effects of nicotine on repair of free radical damaged plasmids by whole-cell protein extracts from adult myocytes. Exposure to a concentration of 25 microM nicotine increased incorporation of (32P)dCTP into damaged plasmids by 16%, and 50 or 100 microM nicotine increased incorporation by 32%. Nicotine did not alter the rate or amount of poly (ADP-ribose) on the major protein acceptor of molecular weight 113-116 kDa. Inhibition of DNA polymerase activity with pyridoxal 5'-phosphate revealed greater plasmid degradation in the presence of nicotine. We conclude that nicotine enhances DNA degradation and the increased repair is a consequence of this greater degradation. PMID- 9737936 TI - Beneficial effect of raxofelast, an hydrophilic vitamin E analogue, in the rat heart after ischemia and reperfusion injury. AB - Several studies report that among the antioxidant agents used to reduce injury after myocardial ischemia/reperfusion, analogues of vitamin E (VE) seem to have a significant efficacy. Raxofelast is a potent antioxidant agent under investigation, structurally related to VE, having an excellent bioavailability and favourable physicochemical properties. We assessed raxofelast in a rat model of myocardial damage induced by 1 h of left coronary artery occlusion followed by 6 h of reperfusion. Myocardial ischemia/reperfusion produced: wide tissue necrosis (50.3+/-10.3%); membrane peroxidation, evaluated by assessing cardiac malondialdehyde (MAL) (87.8+/-15.8 nmol/g tissuev 9.53+/-2.4 nmol/g tissue) and plasma conjugated dienes (CD) (8.73+/-1.86 DeltaABS/mlv 1.61+/-0.45 DeltaABS/ml); endogenous antioxidant wasting [cardiac VE=23.5+/-10.2 nmol/g tissuev 61.4+/-13.4 nmol/g tissue, cardiac reduced glutatione (GSH)=2.15+/-1.23 micromol/g proteinv 7.34+/-0.92 micromol/g protein and cardiac superoxide dismutase (SOD)=8.9+/-4.1 U/mg proteinv 17. 5+/-4.2 U/mg protein]; depressed mean arterial blood pressure (MAP) (61.4+/-5.8 mmHgv 85.3+/-6.2 mmHg); heart rate (HR) (275+/-35 beats/minv 368+/-34 beats/min) and left-ventricular derivative developed force (LV dP/dtmax) (1050+/-187 mmHg/sv 2520+/-194 mmHg/s); and cardiac neutrophil accumulation, evaluated by assessing cardiac myeloperoxidase (MPO) (9.23+/-2.1 U/g tissuev 0.92+/-0.12 U/g tissue). Administration of raxofelast (25, 50 and 100 mg/kg i.p. 5 min after occlusion) limited myocardial necrosis (22.3+/-14.8%P<0. 005, following the highest dose), reduced lipid peroxidation (MAL=43. 5+/-14.7 nmol/g tissueP<0.001 and CD=4.01+/-2.21 DeltaABS/mlP<0.001, following the highest dose), restored the endogenous antioxidants VE (52.8+/-14.2 nmol/g tissueP<0.001, following the highest dose), SOD (14.2+/-2.7 U/mg proteinP<0.001, following the highest dose) and GSH (4.92+/-1.33 micromol/g proteinP<0.005, following the highest dose), improved hemodynamic parameters (MAP=68.1+/-5.3 mmHgP<0.05, HR=317+/-27 beats/minP<0.05, LV dP/dtmax=1427+/-143 mmHg/sP<0.05, following the highest dose) and reduced myocardial neutrophil infiltration (MPO=5.1+/-1.5 U/g tissueP<0.001, following the highest dose). These data suggest that raxofelast could be considered a useful drug to reduce myocardial infarction. PMID- 9737938 TI - Selective activation of alpha1A-adrenergic receptors in neonatal cardiac myocytes is sufficient to cause hypertrophy and differential regulation of alpha1 adrenergic receptor subtype mRNAs. AB - Prolonged stimulation of cardiac alpha1-adrenergic receptors causes myocyte hypertrophy, although the receptor subtypes involved remain controversial. We have used a potent and selective alpha1A agonist, A-61603, to test whether activation of the alpha1A-adrenergic receptor subtype is sufficient to mediate the morphological, biochemical and molecular alterations associated with cardiomyocyte hypertrophy. In neonatal rat cardiomyocyte cultures, 48 h incubation with 50 nm A-61603 led to a marked increase in myocardial cell size that was associated with a significant elevation in the rate of protein synthesis. The increased rate of incorporation of radiolabelled amino acids into protein stimulated by A-61603 was totally abolished by the selective alpha1A antagonist KMD-3213. A-61603 increased ANF secretion three-fold, and ANF mRNA 12 fold above control levels in cardiomyocyte cultures. RNase protection analysis demonstrated a A-61603-mediated two to three-fold increase in alpha1A-adrenergic receptor mRNA with a concomitant 50% decrease in alpha1B mRNA levels by 48 h. Identical responses of differential regulation of alpha1A- and alpha1B-adrenergic receptor mRNA were observed with phenylephrine. Both the stimulation of alpha1A- and repression alpha1B-adrenergic receptor mRNA caused by A-61603 could be abolished by 10-20 nm KMD-3213. The present data provide evidence that selective activation of alpha 1A-adrenergic receptors on cardiomyocytes is sufficient to mediate the phenotypic changes associated with cardiac hypertrophy. In addition, the differential regulation of alpha1A and alpha1B mRNA in response to selective alpha1A-adrenergic receptor stimulation suggests that cross-talk between receptor subtypes may be involved in regulating receptor populations during chronic agonist exposure. PMID- 9737937 TI - Remodelling of cardiac extracellular matrix during beta-adrenergic stimulation: upregulation of SPARC in the myocardium of adult rats. AB - Our objectives were (i) to evaluate the expression of several genes involved in the remodelling of cardiac extracellular matrix (ECM), with a special interest on SPARC (secreted protein acidic and rich in cysteine) a glycoprotein with anti adhesive properties, and (ii) to characterise structural changes in the left (LV) and right (RV) ventricles of rats subjected to continuous beta-adrenergic stimulation. The rats were infused for 3 or 7 days with isoproterenol (ISO, 4 mg/kg/day) or vehicle. Hybridisation analysis was done for SPARC, atrial natriuretic peptide (ANP),alpha2 (I) [COL-I] and alpha1 (III) [COL-III] procollagens, TGF-beta1 and TGF-beta3 mRNA content. Interstitial and perivascular collagen deposition in both ventricles was measured after specific staining. The mean cross-sectional area of LV cardiomyocytes was evaluated by quantitative histomorphometry. ISO provoked an increase of LV mass, and a progressive enlargement of cardiomyocytes: their cross-sectional area raised from 205+/-8 micrometer2 in vehicle-treated animals to 247+/-4 and 296+/-9 micrometer2 after 3 or 7 days of ISO infusion, respectively (P<0.001). SPARC messenger abundance increased by more than 50% in LV and RV, a first evidence of its expression in the myocardium of adult rats. Transcripts of ANP, COL-III, TGF-beta1 and TGF beta3 increased in both ventricles. COL-I transcript increased in LV (75 and 116% on days 3 and 7), but not in RV. In LV, collagen accumulated in the interstitium (2.69+/-0.20v 9. 23+/-0.50% of tissue area for vehicle and ISO 7 days groups, P<0.05) and around coronary arteries (1.04+/-0.11v 4.47+/-0.48% of lumen area for vehicle and ISO 7 days,P<0.05). Cardiac fibrosis was less marked in RV. In conclusion, early expression of SPARC, an anti-adhesive protein, and preferential expression of COL-III, a distensible form of collagen, should increase ECM plasticity and facilitate ventricular remodelling. PMID- 9737939 TI - Refeeding reverses cardiac myosin shifts induced by undernutrition in aged rats: modulation by growth hormone. AB - Growth hormone (GH) was administered (1 mg/day, i.p., 7.5 months) to male Fischer 344 rats, in conjunction with refeeding (RF) after chronic undernutrition (UN), from middle age (17 months old) to senescence (24.5 months old), during which cardiac myosin heavy chain (MHC) profiles were determined by gel-electrophoresis. At 17 months of age, respective MHC-alpha and -beta composition was 74 and 26% in the right ventricle (RV), and 58 and 42% in the left ventricle (LV), of ad libitum-fed controls. At 24.5 months of age, MHC profiles of controls were shifted toward the MHC-beta isoform in both RV (alpha=53%, beta=47%) and LV (alpha=40%, beta=60%), indicating a significant effect of aging on MHC composition in both ventricles. At 17 months of age, 7.5 months of UN likewise resulted in a shift toward the MHC-beta isoform in both RV (alpha=31%, beta=69%) and LV (alpha=22%, beta=78%) as compared to controls, indicating a significant effect of UN in both ventricles. Continued UN into senescence maintained these altered profiles in both ventricles, at 24.5 months of age (RV: alpha=35%, beta=65%; LV: alpha=24%, beta=76%). RF+GH administered from middle age into senescence restored the MHC composition in both ventricles (RV: alpha=57%, beta=43%; LV: alpha=43%, beta=57%), to that of the controls. RF, alone, likewise reversed ventricular MHC composition toward that of MHC-alpha, but appeared to overcompensate (RV: alpha=67%, beta=33%; LV: alpha=46%, beta=54%), surpassing the control and RF+GH profiles, significantly in the RV. These data suggest that GH is a modulator of restoration of cardiac MHC composition, when RF is administered to counter the effects of chronic UN, in the aging rat heart. PMID- 9737940 TI - Augmentation of arterial endothelial cell expression of the plasminogen activator inhibitor type-1 (PAI-1) gene by proinsulin and insulin in vivo. AB - Diabetes mellitus is associated with an increased incidence and greater severity of primary atherosclerosis as well as restenosis after angioplasty for reasons not yet clear. We have shown previously that insulin and proinsulin-typically elevated in blood in patients with type II diabetes-increase plasma activity of plasminogen activator inhibitor type 1 (PAI-1)in vivo. Others have demonstrated that increased PAI-1 activity is associated with coronary heart disease. Accordingly, the present study was performed to identify sites of increased expression of the PAI-1 gene within the vessel wall. Equimolar concentrations of insulin, proinsulin, or vehicle alone as a control, were administered intravenously over 1 h to conscious rabbits that were kept euglycemic throughout by the use of glucose clamping. Within 3 h plasma PAI-1 activity increased from 1.15+/-1.34 to 11.33+/-4.30 AU/ml with insulin (mean+/-s.d., P=0.015) and from 2.83+/-0.74 to 15.43+/-4.70 AU/ml with proinsulin (P=0.035). This was found to be in contrast to the controls where the increase in plasma PAI-1 activity was of lesser degree (2.43+/-1.86 to 6.80+/-1.10 AU/ml, P=n.s., n=4 each). As judged from the results of in situ hybridization, the site of prominent aortic expression of the PAI-1 gene was the endothelium. Furthermore, expression increased further in this site after administration of insulin or proinsulin. As judged from results of immunohistochemistry, PAI-1 protein in the aorta was also prominent in endothelium. These results suggest that "hyper(pro)insulinemia", increases PAI-1 not only in blood but also in arterial endothelium. Thus, attenuation of vasculopathy and especially of restenosis after angioplasty in type II diabetes may be possible with somatic gene therapy targeting PAI-1 expression in endothelial cells. PMID- 9737941 TI - Beta-tropomyosin overexpression induces severe cardiac abnormalities. AB - Tropomyosin, a coiled-coil dimer, stabilizes actin filaments and is central to the control of calcium-regulated striated muscle contraction. Striated muscle specific alpha-tropomyosin is the predominant isoform in cardiac muscle, with low levels of beta-tropomyosin restricted to fetal development in the mouse. To understand the functional role of various tropomyosin isoforms during myofilament activation and regulation in the intact sarcomere, we generated transgenic mice that overexpress striated muscle-specific beta-tropomyosin in the adult heart. Our earlier results succinctly demonstrate that overexpression of beta tropomyosin in the hearts of transgenic mice decreases endogeneous alpha tropomyosin levels while altering diastolic function of the myocardium. To explore further the significance of altering the alpha- to beta-tropomyosin isoform ratio in developing murine myocardium, we generated transgenic mice which express beta-tropomyosin at high levels in the heart. The data show that higher levels of beta-tropomyosin expression are lethal with death ensuing between 10-14 days postnatally. A detailed histological analysis demonstrates that the hearts of these mice exhibit several pathological abnormalities, including thrombus formation in the lumen of both atria and in the subendocardium of the left ventricle. Other changes include atrial enlargement and fibrosis, and diffuse myocytolysis, Physiological analyses using ventricular muscle strip preparations from these mice reveal that both myocardial contraction and relaxation parameters are severely impaired. Thus, these results firmly demonstrate an essential difference in tropomyosin isoform function in physiologically regulating cardiac performance. PMID- 9737942 TI - Angiotensin II, transforming growth factor-beta1 and repair in the infarcted heart. AB - Tissue repair following myocardial infarction (MI) eventuates in fibrous tissue formation at the site of myocyte necrosis. Following a large transmural MI, fibrosis appears remote to the infarct site. This is associated with extensive tissue remodeling that adversely affects ventricular diastolic function. Substances involved in promoting fibrous tissue formation at MI and remote sites are under investigation. Angiotensin II (AngII), generated at sites of repair, has been implicated. However, its regulatory role on fibrous tissue formation remains uncertain. In the present study we sought to determine whether AngII is correlated to transforming growth factor beta 1 (TGF-beta1) expression, a regulator of fibrous tissue formation, at these sites of tissue repair. We studied: (1) localization and expression of angiotensin converting enzyme (ACE), AngII receptors, TGF-beta1 mRNA and its receptors in the infarcted rat heart; and (2) effect of AngII on TGF-beta1 synthesis by chronic blockade of AT1 receptors began at the time of surgery by losartan in rats with MI. Hearts were studied at 4 weeks post-MI. We found: (1) low-density ACE, AngII and TGF-beta1 receptor binding and low mRNA for type I collagen and TGF-beta1 in the normal heart; (2) fibrosis at sites of MI and remote to it, including endocardium and fibrosis of intraventricular septum, interstitial fibrosis of non-infarcted myocardium and fibrosis of visceral pericardium; (3) markedly increased (P<0.01) and colocalized ACE, AngII and TGF-beta1 receptor binding, type I collagen and TGF-beta1 mRNA at MI and remote sites of repair; (4) increased TGF-beta1 concentration (P<0. 01) at these sites; and (5) attenuated TGF-beta1 and type I collagen gene expression (P<0.01) at these sites in rats receiving losartan. These observations suggest locally generated AngII via ATi receptor binding is correlated to TGF-beta1 expression and synthesis at sites of repair and remote sites in the infarcted rat heart. The mechanism responsible for the role of AngII in TGF-beta1 remains to be elucidated. PMID- 9737943 TI - Malate-aspartate shuttle, cytoplasmic NADH redox potential, and energetics in vascular smooth muscle. AB - The effect of inhibition of the malate-aspartate shuttle on the cytoplasmic NADH/NAD ratio and NADH redox state and its corresponding effects on mitochondrial energetics in vascular smooth muscle were examined. Incubation of porcine carotid arteries with 0. 4 mmol amino-oxyacetic acid an inhibitor of glutamate-oxaloacetate transaminase and, hence the malate-aspartate shuttle, inhibited O2 consumption by 21%, decreased the content of phosphocreatine and inhibited activity of the tricarboxylic acid cycle. The rate of glycolysis and lactate production was increased but glucose oxidation was inhibited. These effects of amino-oxyacetic acid were accompanied by evidence of inhibition of the malate-aspartate shuttle and elevation in the cytoplasmic redox potential and NADH/NAD ratio as indicated by elevation of the concentration ratios of the lactate/pyruvate and glycerol-3-phosphate/dihydroxyacetone phosphate metabolite redox couples. Addition of the fatty acid octanoate normalized the adverse energetic effects of malate-aspartate shuttle inhibition. It is concluded that the malate-aspartate shuttle is a primary mode of clearance of NADH reducing equivalents from the cytoplasm in vascular smooth muscle. Glucose oxidation and lactate production are influenced by the activity of the shuttle. The results support the hypothesis that an increased cytoplasmic NADH redox potential impairs mitochondrial energy metabolism. PMID- 9737944 TI - Mechanisms of cardiomyocyte dysfunction in heart failure following myocardial infarction in rats. AB - Available information regarding the cellular and molecular mechanisms for reduced myocardial function after myocardial infarction (MI) is scarce. In rats with congestive heart failure (CHF), we examined cardiomyocytes isolated from the non infarcted region of the left ventricle 6 weeks after ligation of the left coronary artery. Systolic left-ventricular pressure was reduced and diastolic pressure was markedly increased in the CHF-rats. The cardiomyocytes isolated from the CHF-hearts had increased resting length, reduced fractional shortening by 31% and a 34% increase in time to 90% relaxation compared to sham cells (P<0.01 for all). Peak L-type calcium currents were not significantly changed, but peak calcium transients measured with fura-2 were reduced by 19% (P<0.01). Moreover, the decline of the calcium transients as measured by the time constant of a monoexponential function was significantly increased by 26% (P<0.01). We also examined the contribution of the Ca2+-ATPase of the sarcoplasmic reticulum (SR) in the removal of cytosolic Ca2+ during relaxation by superfusing cells with 1 microM thapsigargin that effectively inhibits the Ca2+-ATPase. Relaxation time in CHF-cells was significantly less prolonged when this drug was used (P<0.01). This suggests that other mechanisms, probably the Na+-Ca2+ exchanger, contribute significantly to the relaxation rate in CHF. Simultaneous measurements of fura-2 transients and mechanical shortening did not reveal any alteration in the calcium myofilament sensitivity in CHF. Our study clearly shows reduced shortening and prolonged relaxation in cardiomyocytes isolated from non-infarcted region of the left ventricle in heart failure. Moreover, we were able to relate the observed cardiomyocyte dysfunction to changes in specific steps in the excitation contraction coupling. PMID- 9737946 TI - Calcium responsiveness in canine pacing-induced heart failure. AB - The specific lesion(s) and potential compensatory alterations of excitation contraction coupling in heart failure are not clear in detail. We therefore subjected five dogs to 2-5 weeks of rapid ventricular pacing until heart failure developed. Data obtained from these five dogs with pacing-induced heart failure were compared to data from six healthy controls. Under anesthesia, in situ steady state responses of regional contractile function to intracoronary calcium infusion were established. Maximal calcium-activated regional contractile function in dogs with heart failure was 46% less than in controls; calcium sensitivity was unchanged [pCa50 2.55+/-0.31 v 2.82+/-0.17 (+/-s.d.)]. Our data point to a decrease in maximal calcium-activated force and an unchanged calcium sensitivity if an unchanged calcium transient is assumed, or a compensatory increase in calcium sensitivity of failing myocardium if a decreased calcium transient is assumed. PMID- 9737945 TI - Complexities underlying the quantitative determination of myocardial glucose uptake with 2-deoxyglucose. AB - The quantitative determination of glucose uptake by using 2-deoxy-D-glucose (2 DG) is based on the assumption that 2-deoxyglucose-6-phosphate (2-DG6P) cannot be further metabolized and requires the lumped constant (LC) to equate the kinetic differences in uptake between 2-DG and glucose. We tested whether insulin or epinephrine affect the LC, and quantitated the incorporation of 2-DG6P into glycogen in the isolated working rat heart. Hearts were perfused for 35 min at near physiological workload with Krebs-Henseleit buffer containing glucose (5 mmol/l) plus oleate (0.4 mmol/l, Group 1) with either insulin (1 mU/ml, Group 2), or epinephrine (1 micromol/l, Group 3). In all groups [2-3H] glucose and [U-14C]2 DG (10 microCi each) were present in the perfusate for the first 30 min. In order to estimate the quantitative relationship of glucose and 2-DG uptake and glycogen synthesis from glucose and 2-DG, we perfused hearts with equimolar amounts of glucose and 2-DG (5 mmol each) and either [18F]2-deoxy-2-fluoroglucose plus [2 3H]glucose or [U-14C]glucose plus [1,2-3H]2-DG as tracers. All hearts were freeze clamped for determination of 2-DG accumulation, glycogen, and tracer activity in glycogen. Glucose and 2-DG uptake were similar in the absence of insulin (LC 1.27+/-0.09). In the presence of insulin, 2-DG underestimated glucose uptake (LC 0.61+/-0.02). Epinephrine did not affect the tracer/tracee ratio (LC 1.31+/ 0.09). Incorporation of [U-14C]2-DG into glycogen occurred in all groups (Group 15.38+/-0. 65%, Group 25.72+/-0.59%, Group 32.70+/-0.16% of total tracer uptake.) When equimolar amounts of glucose and 2-DG were present, 2-DG uptake, measured by dynamic assessment of FDG accumulation, significantly decreased over 30 min while glucose uptake remained unchanged. The hearts perfused with [U-14C]glucose and [1,2-3H]2-DG synthesized 39.5+/-7.1 micromol glycogen/g dry/30 min. 2-DG contributed 4.2+/-1.4%. We conclude that insulin and epinephrine have differential effects on the LC, and 2-DG6P is a substrate for glycogen synthesis. PMID- 9737947 TI - Post-infarction heart failure in the rat is associated with distinct alterations in cardiac myocyte molecular phenotype. AB - The myocardial molecular and cellular responses to hemodynamic and other hypertrophic stimuli have been characterized extensively, but less is known of the alterations in gene expression during the evolution of heart failure following myocardial infarction, and specifically those affecting the cardiac myocytes. Therefore, the present study was undertaken to test the hypothesis that post-infarction heart failure and remodeling in the rat is associated with a distinct myocyte molecular phenotype. To address this question, hemodynamic measurements were performed in vivo; and myocytes isolated from the non-infarcted myocardium 1 day, 1 week, and 6 weeks post-coronary artery ligation in post infarct rats and sham controls. Myocyte size, mRNA levels for immediate early genes, contractile proteins, and sarcoplasmic reticulum Ca2+-ATPase (SERCA) and phospholamban were assayed by Northern analyses, and SERCA and phospholamban proteins were examined by Western blotting. Hemodynamic evidence of heart failure was present at all post-infarct time points. Myocyte size was increased significantly at 6 weeks. c-myc expression was increased at 1 day and 1 week in the infarcted rats, but returned to baseline by 6 weeks. Atrial natriuretic peptide and VEGF mRNAs were elevated at 1 and 6 weeks. Both beta-myosin heavy chain and skeletal alpha-actin expression were increased at all post-MI time points. In contrast, neither changes in the expression of the calcium-handling proteins (SERCA and phospholamban) were not observed, nor was there a change in TGFbeta1 or TGFbeta3. These results demonstrate that in rats with post-MI heart failure, there was an immediate induction of the fetal/embryonic transcriptional gene program which preceded myocyte hypertrophy and appeared to persist longer than in pressure-overload models. In further contrast to pressure-overload, expression of sarcoplasmic reticulum Ca2+-ATPase and phospholamban, was not altered despite a comparable degree of cellular hypertrophy and more severe hemodynamic decompensation. These findings suggest that there may be important differences in the regulatory mechanisms underlying these two forms of myocardial hypertrophy and heart failure. PMID- 9737948 TI - Energy-dependent transport of calcium to the extracellular space during acute ischemia of the rat heart. AB - OBJECTIVE: Acute ischemia is associated with rapidly decreasing contractility and Ca2+-transients. Diastolic intracellular Ca2+, however, only mildly increases until development of contracture. The purpose of this study was to investigate whether changes of cellular calcium handling during the early phase of ischemia are associated with active sarcolemmal calcium transport. METHODS: Changes of extracellular concentration of calcium ([Ca2+]o) and tetramethylammonium ([TMA+]o), to estimate extracellular space, were simultaneously measured with ion specific electrodes in the globally ischemic rat heart. The magnitude and direction of sarcolemmal calcium transport were calculated from [Ca2]o corrected for changed extracellular water content. Energy dependence of sarcolemmal calcium transport was investigated by application of iodoaceticacid (IAA) to inhibit anaerobic glycolysis, and the involvement of the sarcoplasmic reticulum (SR) was studied by application of thapsigargin. The effect of anoxia and thapsigargin on cytosolic and SR calcium was studied in isolated myocytes with the fluorescent indicator indo-1. RESULTS: [Ca2+]o increased and extracellular space gradually decreased in the ischemic intact heart. During the first 7 min, the increase of [Ca2+]o was associated with net outward transport of calcium. Subsequently, net re-uptake occurred. IAA completely abolished outward transport and influx was accelerated and enhanced. Application of thapsigargin attenuated outward transport. In electrically-stimulated myocytes, anoxia caused little change of diastolic calcium and depletion of SR. Thapsigargin reduced both calcium transient amplitude and SR calcium without affecting diastolic calcium. During three successive short episodes of ischemia/reperfusion (preconditioning), outward transport of calcium progressively decreased. CONCLUSION: During the early phase of global ischemia, energy dependent transport of calcium to the extracellular space occurs. At least part of this calcium originates from SR. During the later stage of ischemia, re-uptake of calcium occurs, which is associated with development of contracture. PMID- 9737949 TI - Difference in the mechanisms for compensating ischemic acidosis in diabetic rat hearts. AB - To elucidate the difference in the mechanisms for alkalization during ischemic acidosis between diabetic and non-diabetic hearts, intracellular pH (pHi) was measured by phosphorus-31 magnetic resonance spectroscopy. Diabetes was induced by the injection of streptozotocin. The accumulation of proton ion (DeltaH+) during 15 min global ischemia at 37 degreesC was calculated from pH i. There were no significant differences in DeltaH+ between diabetic (DM: 0. 54+/-0.03 micromol/l,n=6; mean+/-s.e.m.) and non-DM hearts (0.57+/-0.04, n=6), when perfused with bicarbonate buffer. However, perfusion with HEPES buffer revealed a significant increase of DeltaH+ in DM (0.85+/-0.07, n=5) compared with non-DM (0.61+/-0.06, n=5P<0.05). On the contrary, the addition of a Na+/H+ exchange inhibitor (EIPA; 1 micromol/l) to bicarbonate buffer significantly increased DeltaH+ in non-DM (1.09+/-0.10, n=4) compared with DM (0.71+/-0.03, n=5P<0.01). Perfusion with HEPES buffer and EIPA equally increased DeltaH+ in both groups (DM 1.13+/-0.13, n=4; non-DM 1.15+/-0.14, n=4). Thus, the activity of Na+/H+ exchanger during ischemic acidosis, assessed as the increase of DeltaH+ induced by addition of EIPA to bicarbonate buffer, was higher in non-DM (0.52) than DM (0.17). In contrast, the contribution of bicarbonate-dependent systems evaluated by the deference of DeltaH+ between the bicarbonate buffer and the HEPES buffer was markedly bigger in DM (0.31) than non-DM (0.04). These results indicate that Na+/H+ exchange is a major mechanism to compensate ischemic acidosis in non-DM hearts, whereas bicarbonate-dependent systems compensate the depressed activity of Na+/H+ exchange in DM. PMID- 9737950 TI - Cytochalasin D alters kinetics of Ca2+ transient in rat ventricular cardiomyocytes: an effect of altered actin cytoskeleton? AB - The effects of cytochalasin D, a specific F-actin depolymerizing agent, on Ca2+ transients in rat ventricular cardiomyocytes were investigated. Cytochalasin D (20 microM) significantly slowed decay of Ca2+ transients (tau decay control cells=28.1+/-1.3, n=28tau decay=47.3+/-2.8 ms, n=20, P<0.001). The rising phase of Ca2+ transients was also significantly slower in cytochalasin D treated cells (tau rise=5.1+/-0.6 ms, n=17nu tau rise in control cells=3.6+/-0.2, n=21,P<0.01). Phalloidin (100 microM), an F-actin stabilizer, prevented cytochalasin D-induced alterations of Ca2+ transient kinetics. The cytochalasin D effect was not related to the l-type Ca2+ current since the current density and kinetics were not altered by the drug. We conclude that integrity of F-actin-based cytoskeleton is an important factor for sarcoplasmic reticulum function. PMID- 9737951 TI - Regulated diversity of heparan sulfate. PMID- 9737952 TI - Localization in the II-III loop of the dihydropyridine receptor of a sequence critical for excitation-contraction coupling. AB - Skeletal and cardiac muscles express distinct isoforms of the dihydropyridine receptor (DHPR), a type of voltage-gated Ca2+ channel that is important for excitation-contraction (EC) coupling. However, entry of Ca2+ through the channel is not required for skeletal muscle-type EC coupling. Previous work (Tanabe, T., Beam, K. G., Adams, B. A., Niidome, T., and Numa, S. (1990) Nature 346, 567-569) revealed that the loop between repeats II and III (II-III loop) is an important determinant of skeletal-type EC coupling. In the present study we have further dissected the regions of the II-III loop critical for skeletal-type EC coupling by expression of cDNA constructs in dysgenic myotubes. Because Ser687 of the skeletal II-III loop has been reported to be rapidly phosphorylated in vitro, we substituted this serine with alanine, the corresponding cardiac residue. This alanine-substituted skeletal DHPR retained the ability to mediate skeletal-type EC coupling. Weak skeletal-type EC coupling was produced by a chimeric DHPR, which was entirely cardiac except for a small amount of skeletal sequence (residues 725-742) in the II-III loop. Skeletal-type coupling was stronger when both residues 725-742 and adjacent residues were skeletal (e.g. a chimera containing skeletal residues 711-765). However, residues 725-742 appeared to be critical because skeletal-type coupling was not produced either by a chimera with skeletal residues 711-732 or by one with skeletal residues 734-765. PMID- 9737953 TI - Phosphorylation is not required for dynamin-dependent endocytosis of a truncated mutant opioid receptor. AB - Opioid receptors are regulated within minutes after activation by G protein coupled receptor kinase-mediated phosphorylation and dynamin-dependent endocytosis. We addressed the question of whether phosphorylation is required for opioid receptor endocytosis by examining a functional, truncated mutant delta opioid receptor (DOR344T), which is missing phosphorylation sites located in the carboxyl-terminal cytoplasmic domain. DOR344T receptors expressed in Chinese hamster ovary cells remained predominantly in the plasma membrane, even in the presence of saturating concentrations of agonist, consistent with previous studies demonstrating strongly inhibited endocytosis of truncated receptors in this cell type. In marked contrast, DOR344T receptors expressed at similar levels in human embryonal kidney (HEK) 293 cells exhibited rapid, ligand-induced internalization either in the presence of peptide (DADLE) or alkaloid (etorphine) agonist. Quantitative assays using ELISA and flow cytometric techniques indicated that DOR344T receptors were endocytosed in HEK293 cells with similarly rapid kinetics as full-length DOR (t1/2 < 10 min), and both full-length DOR and DOR344T mutant receptors were endocytosed by a dynamin-dependent mechanism involving clathrin-coated pits. Nevertheless, DOR344T receptors failed to undergo any detectable constitutive or agonist-induced phosphorylation in the same cells in which dynamin-dependent endocytosis was observed. These findings establish the first example of a G protein-coupled receptor that does not require phosphorylation to undergo dynamin-dependent endocytosis, and they suggest that significant cell type-specific differences exist in the biochemical requirements for ligand-induced concentration of opioid receptors in clathrin-coated pits. PMID- 9737954 TI - Kinetic analysis of the mechanism and specificity of protein-disulfide isomerase using fluorescence-quenched peptides. AB - Protein-disulfide isomerase (PDI) is an abundant folding catalyst in the endoplasmic reticulum of eukaryotic cells. PDI introduces disulfide bonds into newly synthesized proteins and catalyzes disulfide bond isomerizations. We have synthesized a library of disulfide-linked fluorescence-quenched peptides, individually linked to resin beads, for two purposes: 1) to probe PDI specificity, and 2) to identify simple, sensitive peptide substrates of PDI. Using this library, beads that became rapidly fluorescent by reduction by human PDI were selected. Amino acid sequencing of the bead-linked peptides revealed substantial similarities. Several of the peptides were synthesized in solution, and a quantitative characterization of pre-steady state kinetics was carried out. Interestingly, a greater than 10-fold difference in affinity toward PDI was seen for various substrates of identical length. As opposed to conventional PDI assays involving larger polypeptides, the starting material for this assay is homogenous. It is furthermore simple and highly sensitive (requires less than 0.5 microgram of PDI/assay) and thus opens the possibility for quantitative determination of PDI activity and specificity. PMID- 9737955 TI - Proteins of newly isolated mutants and the amino-terminal proline are essential for ubiquitin-proteasome-catalyzed catabolite degradation of fructose-1,6 bisphosphatase of Saccharomyces cerevisiae. AB - Addition of glucose to cells of the yeast Saccharomyces cerevisiae growing on a non-fermentable carbon source leads to selective and rapid degradation of fructose-1,6-bisphosphatase. This so called catabolite inactivation of the enzyme is brought about by the ubiquitin-proteasome system. To identify additional components of the catabolite inactivation machinery, we isolated three mutant strains, gid1, gid2, and gid3, defective in glucose-induced degradation of fructose-1,6-bisphospha-tase. All mutant strains show in addition a defect in catabolite inactivation of three other gluconeogenic enzymes: cytosolic malate dehydrogenase, isocitrate lyase, and phosphoenolpyruvate carboxykinase. These findings indicate a common mechanism for the inactivation of all four enzymes. The mutants were also impaired in degradation of short-lived N-end rule substrates, which are degraded via the ubiquitin-proteasome system. Site-directed mutagenesis of the amino-terminal proline residue yielded fructose-1,6 bisphosphatase forms that were no longer degraded via the ubiquitin-proteasome pathway. All amino termini other than proline made fructose-1,6-bisphosphatase inaccessible to degradation. However, the exchange of the amino-terminal proline had no effect on the phosphorylation of the mutated enzyme. Our findings suggest an essential function of the amino-terminal proline residue for the degradation process of fructose-1,6-bisphosphatase. Phosphorylation of the enzyme was not necessary for degradation to occur. PMID- 9737957 TI - c-Fos degradation by the proteasome. An early, Bcl-2-regulated step in apoptosis. AB - c-Fos is a transcription factor that promotes cell growth, differentiation, and transformation. We found that c-Fos was degraded when WEHI7.2 mouse lymphoma cells were induced to undergo apoptosis with the calcium ATPase inhibitor, thapsigargin, or the glucocorticoid hormone, dexamethasone. The degradation of c Fos preceded caspase-3 activation and apoptotic nuclear chromatin condensation and was inhibited by the proteasome inhibitors MG132, N-acetyl-leucyl-leucyl norleucinal, and lactacystin. Stable transfection of WEHI7.2 cells with a mutant form of c-Fos that was not degraded by the proteasome inhibited apoptosis. Also, overexpression of Bcl-2 in WEHI7.2 cells blocked c-Fos degradation and inhibited apoptosis. The results indicate that proteasome-mediated degradation of c-Fos is an early, Bcl-2-regulated step in apoptosis induction by thapsigargin and dexamethasone. These findings suggest that c-Fos may have a protective action that is eliminated by proteasome-mediated degradation and preserved by Bcl-2. PMID- 9737958 TI - Calcium release-activated calcium current (ICRAC) is a direct target for sphingosine. AB - Whole cell patch-clamp recordings were made to study the regulation of the store operated calcium release-activated calcium current (ICRAC) by metabolites involved in the sphingomyelin pathway in RBL-2H3 cells. Sphingosine, a regulator of cell growth, inhibits ICRAC completely within 200 s and independently from conversion to either sphingosine 1-phosphate or ceramide. Structural analogs of sphingosine, including N,N-dimethylsphingosine, DL-threo-dihydrosphingosine, and N-acetylsphingosine (C2-ceramide) also block ICRAC. This effect is always accompanied by an elevation of whole cell membrane capacitance. These sphingolipids appear, therefore, to accumulate in the plasma membrane and directly block ICRAC channels. Sphingosylphosphorylcholine also increases capacitance but does not inhibit ICRAC, demonstrating structural specificity and that the elevation of capacitance is necessary but not sufficient for block. Nerve growth factor, which is known to break down sphingomyelin, inhibits ICRAC, and this inhibition can be antagonized by reducing sphingosine production with L cycloserine, suggesting that ICRAC is a physiologically relevant and direct target of sphingosine. We propose that sphingosine directly blocks ICRAC, suggesting that the sphingomyelin pathway is involved in ICRAC regulation. PMID- 9737956 TI - Detection of a conserved alpha-helix in the kinase-docking region of the aspartate receptor by cysteine and disulfide scanning. AB - The transmembrane aspartate receptor of Escherichia coli and Salmonella typhimurium propagates extracellular signals to the cytoplasm, where its cytoplasmic domain regulates the histidine kinase, CheA. Different signaling states of the cytoplasmic domain modulate the kinase autophosphorylation rate over at least a 100-fold range. Biochemical and genetic studies have implicated a specific region of the cytoplasmic domain, termed the signaling subdomain, as the region that transmits regulation from the receptor to the kinase. Here cysteine and disulfide scanning are applied to the N-terminal half of the signaling subdomain to probe its secondary structure, solvent exposure, and protein-protein interactions. The chemical reactivities of the scanned cysteines exhibit the characteristic periodicity of an alpha-helix with distinct solvent-exposed and buried faces. This helix, termed alpha7, ranges approximately from residue 355 through 386. Activity measurements probing the effects of cysteine substitutions in vivo and in vitro reveal that both faces of helix alpha7 are critical for kinase activation, while the buried face is especially critical for kinase down regulation. Disulfide scanning of the region suggests that helix alpha7 is not in direct contact with its symmetric partner (alpha7') from the other subunit; presently, the structural element that packs against the buried face of the helix remains unidentified. Finally, a novel approach termed "protein interactions by cysteine modification" indicates that the exposed C-terminal face of helix alpha7 provides an essential docking site for the kinase CheA or for the coupling protein CheW. PMID- 9737959 TI - Chromatin structure and transcriptional control elements of the erythroid Kruppel like factor (EKLF) gene. AB - Erythroid Kruppel-like factor (EKLF) is a red cell-specific transcription factor whose activity is critical for the switch in expression from fetal to adult beta globin during erythroid ontogeny. We have examined its own regulation using a number of approaches. First, the EKLF transcription unit is in an open chromatin configuration in erythroid cells. Second, in vivo transfection assays demonstrate that the more distal of the two erythroid-specific DNase-hypersensitive sites behaves as an enhancer. Although this conserved element imparts high level transcription to a heterologous promoter in all lines examined, erythroid specificity is retained only when it is fused to the proximal EKLF promoter, which contains an important GATA site. Third, extensive mutagenesis of this enhancer element has delimited its in vivo activity to a core region of 49 base pairs. Finally, in vitro footprint and gel shift assays demonstrate that three distinct DNA binding activities in erythroid cell extracts individually interact with three short sequences within this core enhancer element. These analyses reveal that high level erythroid expression of EKLF relies on the interplay between conserved proximal and distal promoter elements that alter chromatin structure and likely provide a target for genetic control via extracellular induction pathways. PMID- 9737960 TI - Ribosomal P-protein stalk function is targeted by sordarin antifungals. AB - Sordarin derivatives are remarkably selective inhibitors of fungal protein synthesis. Available evidence points to a binding site for these inhibitors on elongation factor 2, but high affinity binding requires the presence of ribosomes. The gene mutated in one of the two isolated complementation groups of Saccharomyces cerevisiae mutants resistant to the sordarin derivative GM193663 has now been identified. It is RPP0, encoding the essential protein of the large ribosomal subunit stalk rpP0. Resistant mutants are found to retain most of the binding capacity for the drug, indicating that mutations in rpP0 endow the ribosome with the capacity to perform translation elongation in the presence of the inhibitor. Other proteins of the ribosomal stalk influence the expression of resistance, pointing to a wealth of interactions between stalk components and elongation factors. The involvement of multiple elements of the translation machinery in the mode of action of sordarin antifungals may explain the large selectivity of these compounds, even though the individual target components are highly conserved proteins. PMID- 9737961 TI - Mechanism of reductive activation of potato tuber ADP-glucose pyrophosphorylase. AB - The potato tuber (Solanum tuberosum L.) ADP-glucose pyrophosphorylase activity is activated by a incubation with ADP-glucose and dithiothreitol or by ATP, glucose- 1-phosphate, Ca2+, and dithiothreitol. The activation was accompanied by the appearance of new sulfhydryl groups as determined with 5, 5'-dithiobis(2 nitrobenzoic acid). By analyzing the activated and nonactivated enzymes on SDS polyacrylamide gel electrophoresis under nonreducing conditions, it was found that an intermolecular disulfide bridge between the small subunits of the potato tuber enzyme was reduced during the activation. Further experiments showed that the activation was mediated via a slow reduction and subsequent rapid conformational change induced by ADP-glucose. The activation process could be reversed by oxidation with 5, 5'-dithiobis(2-nitrobenzoic acid). Incubation with ADP-glucose and dithiothreitol could reactivate the oxidized enzyme. Chemical modification experiments with [14C]iodoacetic acid and 4-vinylpyridine determined that the intermolecular disulfide bridge was located between Cys12 of the small subunits of the potato tuber enzyme. Mutation of Cys12 in the small subunit into either Ala or Ser eliminated the requirement of DTT on the activation and prevented the formation of the intermolecular disulfide of the potato tuber enzyme. The mutants had instantaneous activation rates as the wild-type in the reduced state. A two-step activation model is proposed. PMID- 9737962 TI - Unidirectional steady state rates of central metabolism enzymes measured simultaneously in a living plant tissue. AB - The unidirectional steady state reaction rates of several enzymes and metabolic fluxes of distinct processes were measured simultaneously in hypoxic maize root tips using two-dimensional phosphorus NMR exchange spectroscopy. A single spectrum monitors ATP synthesis and hydrolysis as well as the activities of four enzymes involved in key pathways of central metabolism: UDP-glucose pyrophosphorylase, phosphoglucomutase, hexose-phosphate isomerase, and enolase. The corresponding unidirectional reaction rates and net metabolic fluxes were calculated from spectral intensities. This method provides a unique picture, at enzyme resolution, of how metabolism reacts in a concerted fashion to changes in external parameters such as temperature and oxygen concentration. By increasing hypoxia via an increase in temperature, we measured the expected increase in glycolysis through enolase activity while total ATP synthesis settled. At the same time, we observed a net flux through phosphoglucomutase and UDP-glucose pyrophosphorylase toward carbohydrate synthesis. This result is discussed in relation to the current hypothesis on the turnover of cell walls and sucrose. This reaction also produces a net flux of pyrophosphate, which is needed by pyrophosphate:fructose-6-phosphate 1-phosphotransferase to work as a glycolytic enzyme. PMID- 9737963 TI - Redox potential controls the structure and DNA binding activity of the paired domain. AB - Pax proteins are transcriptional regulators controlling a variety of cell fates during animal development. This role depends on the intact function of the paired (Prd) domain that is able to recognize specific DNA sequences. The Prd domain is composed of two distinct helix-turn-helix subdomains, PAI and RED. Molecular functions of Pax proteins are subjected to different levels of regulation involving both pre-translational and post-translational mechanisms. By using Pax 5 and Pax-8 recombinant proteins, we demonstrate that the binding activity of the Prd domain is regulated through the oxidation/reduction of conserved cysteine residues. Mass spectrometry analysis and mutagenesis experiments demonstrate that the redox regulation is accomplished through the reversible formation of an intramolecular disulfide bridge involving the cysteines present in the PAI subdomain, whereas the RED subdomain appears quite insensitive to redox potential. Circular dichroism experiments indicate that only the reduced form of the Prd domain is able to undergo the proper conformational change necessary for sequence-specific DNA binding. Nuclear extracts from different cell lines contain an activity that is able to reduce the Paired domain and, therefore, to control the DNA binding activity of this protein. Immunodepletion of nuclear extracts demonstrate that the protein Ref-1 contributes to the redox regulation of the Prd DNA binding activity. Given the modular nature of the Prd domain and the independent DNA binding specificity of the PAI and RED subdomains, we propose that this control mechanism should be involved in "switching" among different DNA sequences and therefore different target genes. PMID- 9737964 TI - Changing the nature of the initial chaperonin capture complex influences the substrate folding efficiency. AB - For the chaperonin substrates, rhodanese, malate dehydrogenase (MDH), and glutamine synthetase (GS), the folding efficiencies, and the lifetimes of folding intermediates were measured with either the nucleotide-free GroEL or the activated ATP.GroEL.GroES chaperonin complex. With both nucleotide-free and activated complex, the folding efficiency of rhodanese and MDH remained high over a large range of GroEL to substrate concentration ratios (up to 1:1). In contrast, the folding efficiency of GS began to decline at ratios lower than 8:1. At ratios where the refolding yields were initially the same, only a relatively small increase (1.6-fold) in misfolding kinetics of MDH was observed with either the nucleotide-free or activated chaperonin complex. For rhodanese, no change was detected with either chaperonin complex. In contrast, GS lost its ability to interact with the chaperonin system at an accelerated rate (8-fold increase) when the activated complex instead of the nucleotide-free complex was used to rescue the protein from misfolding. Our data demonstrate that the differences in the refolding yields are related to the intrinsic folding kinetics of the protein substrates. We suggest that the early kinetic events at the substrate level ultimately govern successful chaperonin-substrate interactions and play a crucial role in dictating polypeptide flux through the chaperonin system. Our results also indicate that an accurate assessment of the transient properties of folding intermediates that dictate the initial chaperonin-substrate interactions requires the use of the activated complex as the interacting chaperonin species. PMID- 9737965 TI - Free radical intermediates of phenytoin and related teratogens. Prostaglandin H synthase-catalyzed bioactivation, electron paramagnetic resonance spectrometry, and photochemical product analysis. AB - Phenytoin and related xenobiotics can be bioactivated by embryonic prostaglandin H synthase (PHS) to a teratogenic free radical intermediate. The mechanism of free radical formation was evaluated using photolytic oxidation with sodium persulfate and by EPR spectrometry. Characterization of the products by mass spectrometry suggested that phenytoin photolyzes to a nitrogen-centered radical that rapidly undergoes ring opening to form a carbon-centered radical. PHS-1 was incubated with teratogen (phenytoin, mephenytoin, trimethadione, phenobarbital, and major metabolites) or its vehicle and the free radical spin trap alpha-phenyl N-t-butylnitrone, and incubations were analyzed by EPR spectrometry. There was no alpha-phenyl-N-t-butylnitrone radical adduct in control incubations. For phenytoin, a putative unstable nitrogen-centered radical adduct and a stable carbon-centered radical adduct were detected. Free radical spin adducts also were detected for all other teratogens and metabolites except carbamazepine. The PHS inhibitor eicosatetraynoic acid abolished the free radical EPR signal. Incubation of 2'-deoxyguanosine with phenytoin and PHS-1 resulted in a 5-fold increase in its oxidation to 8-hydroxy-2'-deoxyguanosine. This is the first direct chemical evidence for PHS-catalyzed bioactivation of phenytoin and related teratogens to a free radical intermediate that initiates DNA oxidation, which may constitute a common molecular mechanism of teratologic initiation. PMID- 9737966 TI - Potential regulation of Ste20 function by the Cln1-Cdc28 and Cln2-Cdc28 cyclin dependent protein kinases. AB - The activity of the Saccharomyces cerevisiae pheromone signal transduction pathway is regulated by Cln1/2-Cdc28 cyclin-dependent kinase. High level expression of CLN2 can repress activation of the pathway by mating factor or by deletion of the alpha-subunit of the heterotrimeric G-protein. We now show that CLN2 overexpression can also repress FUS1 induction if the signaling pathway is activated at the level of the beta-subunit of the G-protein (STE4) but not when activated at the level of downstream kinases (STE20 and STE11) or at the level of the transcription factor STE12. This epistatic analysis indicates that repression of pheromone signaling pathway by Cln2-Cdc28 kinase takes place at a level around STE20. In agreement with this, a marked reduction in the electrophoretic mobility of the Ste20 protein is observed at the time in the cell cycle of maximal expression of CLN2. This mobility change is constitutive in cells overexpressing CLN2 and absent in cells lacking CLN1 and CLN2. These changes in electrophoretic mobility correlate with repression of pheromone signaling and suggest Ste20 as a target for repression of signaling by G1 cyclins. Two morphogenic pathways for which Ste20 is essential, pseudohyphal differentiation and haploid-invasive growth, also require CLN1 and CLN2. Together with the previous observation that Cln1 and Cln2 are required for the function of Ste20 in cytokinesis, this suggests that Cln1 and Cln2 regulate the biological activity of Ste20 by promoting morphogenic functions, while inhibiting the mating factor signal transduction function. PMID- 9737967 TI - Characterization of the recombinant MutY homolog, an adenine DNA glycosylase, from yeast Schizosaccharomyces pombe. AB - The mutY homolog (SpMYH) gene from a cDNA library of Schizosaccharomyces pombe encodes a protein of 461 amino acids that displays 28 and 31% identity to Escherichia coli MutY and human MutY homolog (MYH), respectively. Expressed SpMYH is able to complement an E. coli mutY mutant to reduce the mutation rate. Similar to E. coli MutY protein, purified recombinant SpMYH expressed in E. coli has adenine DNA glycosylase and apurinic/apyrimidinic lyase activities on A/G- and A/7,8-dihydro-8-oxoguanine (8-oxoG)-containing DNA. However, both enzymes have different salt requirements and slightly different substrate specificities. SpMYH has greater glycosylase activity on 2-aminopurine/G and A/2-aminopurine but weaker activity on A/C than E. coli MutY. Both enzymes also have different substrate binding affinity and catalytic parameters. Although SpMYH has great affinity to A/8-oxoG-containing DNA as MutY, the binding affinity to A/G containing DNA is substantially lower for SpMYH than MutY. SpMYH has similar reactivity to both A/G- and A/8-oxoG-containing DNA; however, MutY cleaves A/G containing DNA about 3-fold more efficiently than it does A/8-oxoG-containing DNA. Thus, SpMYH is the functional eukaryotic MutY homolog responsible for reduction of 8-oxoG mutational effect. PMID- 9737968 TI - Interaction of actin monomers with Acanthamoeba actophorin (ADF/cofilin) and profilin. AB - Acanthamoeba actophorin is a member of ADF/cofilin family that binds both actin monomers and filaments. We used fluorescence anisotropy to study the interaction of actin monomers with recombinant actophorin labeled with rhodamine on a cysteine substituted for Serine-88. Labeled actophorin retains its affinity for actin and ability to reduce the low shear viscosity of actin filaments. At physiological ionic strength, actophorin binds Mg-ADP-actin monomers (Kd = 0.1 microM) 40 times stronger than Mg-ATP-actin monomers. When bound to actin monomers, actophorin has no effect on elongation at either end of actin filaments by Mg-ATP-actin and slightly increases the rate of elongation at both ends by Mg ADP-actin. Thus actophorin does not sequester actin monomers. Sedimentation equilibrium ultracentrifugation shows that actophorin and profilin compete for binding actin monomers. Actophorin and profilin have opposite effects on the rate of exchange of nucleotide bound to actin monomers. Despite the high affinity of actophorin for ADP-actin, physiological concentrations of profilin overcome the inhibition of ADP exchange by actophorin. Profilin rapidly recycles ADP-actin back to the profilin-ATP-actin pool ready for elongation of actin filaments. PMID- 9737969 TI - Inhibition of trypanosomal cysteine proteinases by their propeptides. AB - The ability of the prodomains of trypanosomal cysteine proteinases to inhibit their active form was studied using a set of 23 overlapping 15-mer peptides covering the whole prosequence of congopain, the major cysteine proteinase of Trypanosoma congolense. Three consecutive peptides with a common 5-mer sequence YHNGA were competitive inhibitors of congopain. A shorter synthetic peptide consisting of this 5-mer sequence flanked by two Ala residues (AYHNGAA) also inhibited purified congopain. No residue critical for inhibition was identified in this sequence, but a significant improvement in Ki value was obtained upon N terminal elongation. Procongopain-derived peptides did not inhibit lysosomal cathepsins B and L but did inhibit native cruzipain (from Dm28c clone epimastigotes), the major cysteine proteinase of Trypanosoma cruzi, the proregion of which also contains the sequence YHNGA. The positioning of the YHNGA inhibitory sequence within the prosegment of trypanosomal proteinases is similar to that covering the active site in the prosegment of cysteine proteinases, the three-dimensional structure of which has been resolved. This strongly suggests that trypanosomal proteinases, despite their long C-terminal extension, have a prosegment that folds similarly to that in related mammal and plant cysteine proteinases, resulting in reverse binding within the active site. Such reverse binding could also occur for short procongopain-derived inhibitory peptides, based on their resistance to proteolysis and their ability to retain inhibitory activity after prolonged incubation. In contrast, homologous peptides in related cysteine proteinases did not inhibit trypanosomal proteinases and were rapidly cleaved by these enzymes. PMID- 9737970 TI - Molecular cloning and characterization of the human and mouse UDP-glucose dehydrogenase genes. AB - The enzyme UDP-glucose dehydrogenase (Udpgdh) (EC 1.1.1.22) converts UDP-glucose to UDP-glucuronate, a critical component of the glycosaminoglycans, hyaluronan, chondroitin sulfate, and heparan sulfate. Although Udpgdh is a comparatively well characterized enzyme, no vertebrate genes encoding this enzyme have been reported to date. We report the cloning and characterization of the human and mouse UDP glucose dehydrogenase genes. Mouse and human cDNAs predicted proteins of 493 and 494 amino acids, 24-25 residues longer at their carboxyl termini than the previously reported bovine Udpgdh sequence. The mouse Ugdh gene is composed of 10 exons, spanning 15 kilobases. Northern analyses indicated widespread expression of the gene in embryo and adult. Through interspecific backcross analyses, we localized the Ugdh gene to mouse chromosome 5 at approximately 39 centimorgans, suggesting that the human UGDH gene is localized to chromosome 4p13-15. Results from Southern analyses strongly suggest that Udpgdh is encoded by a single gene in the mouse. Transfection of mouse Ugdh expression vectors led to an increase in detectable Udpgdh activity in mammalian cells. Preliminary expression studies indicated that proinflammatory cytokines, such as interleukin 1beta, can substantially increase the expression of human UGDH in cultured human fibroblasts, suggesting that glycosaminoglycan biosynthesis may be partly regulated by the availability of activated UDP-glucuronate, as determined by relative Udpgdh expression levels. PMID- 9737971 TI - Tetranucleotide GGGA motif in primary RNA transcripts. Novel target site for antisense design. AB - Selecting effective antisense target sites on a given mRNA molecule constitutes a major problem in antisense therapeutics. By trial-and-error, only 1 in 18 (6%) of antisense oligonucleotides designed to target the primary RNA transcript of tumor necrosis factor-alpha (TNF-alpha) strongly inhibited TNF-alpha synthesis. Subsequent studies showed that the area in RNA targeted by antisense oligonucleotides could be moved effectively 10-15 bases in either direction from the original area. We observed that only molecules that incorporated a tetranucleotide motif TCCC (complementary to GGGA on RNA) yielded potent antisense oligonucleotides against TNF-alpha. A comprehensive literature survey showed that this motif is unwittingly present in 48% of the most potent antisense oligonucleotides reported in the literature. This finding was prospectively used to predict the sequences of additional antisense oligonucleotides for the rat TNF alpha primary RNA transcript. Over 50% of antisense constructs (13 of 22) containing the TCCC motif were found to effectively inhibit TNF-alpha synthesis. Marked reductions in mRNA were also observed. This motif was found to be most effective when targeting introns in the primary RNA transcript, suggesting a nuclear localization for the antisense action. Predicting target sites based on the presence of this motif in primary RNA transcripts should be of value in the development on new antisense pharmacotherapy. PMID- 9737972 TI - Disulfide bonding and cysteine accessibility in the alpha-amino-3-hydroxy-5 methylisoxazole-4-propionic acid receptor subunit GluRD. Implications for redox modulation of glutamate receptors. AB - Redox agents elicit a wide variety of effects on the ligand affinity and channel properties of ionotropic glutamate receptors and have been proposed as potential therapeutic agents for neuropathological processes. One such effect is the dithiothreitol (DTT)-induced increase in agonist affinity of certain ionotropic glutamate receptors (GluRs), presumably due to reduction of a disulfide bridge formed between cysteine residues conserved among all GluRs. Using biochemical techniques, this disulfide is shown to exist in the ligand-binding domain of the alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) receptor subunit GluRD, although GluRD homomeric receptors are not modulated by DTT. The disulfide is inaccessible to DTT, explaining the insensitivity of the intact receptor. Single mutants C260S and C315S show a 2-3-fold higher ligand affinity than wild type, as observed for several intact GluRs, indicating that the affinity switch is completely contained within the ligand-binding domain. Also, mutants lacking the native disulfide show non-native oligomerization and dramatically reduced specific activity. These facts suggest that the disulfide bridge is required for the stability of the ligand-binding domain, explaining its conservation. A third cysteine residue in the ligand-binding domain exists as a free thiol, partially sequestered in a hydrophobic environment. These results provide a framework for interpreting a variety of GluR redox modulatory phenomena. PMID- 9737973 TI - Platelet-derived growth factor inhibits insulin stimulation of insulin receptor substrate-1-associated phosphatidylinositol 3-kinase in 3T3-L1 adipocytes without affecting glucose transport. AB - Phosphatidylinositol 3-kinase (PI3K) activation is necessary for insulin responsive glucose transporter (GLUT4) translocation and glucose transport. Insulin and platelet-derived growth factor (PDGF) stimulate PI3K activity in 3T3 L1 adipocytes, but only insulin is capable of stimulating GLUT4 translocation and glucose transport. We found that PDGF causes serine/threonine phosphorylation of insulin receptor substrate 1 (IRS-1) in 3T3-L1 cells, measured by altered mobility on SDS-polyacrylamide gel, and this leads to a decrease in insulin stimulated tyrosine phosphorylation of IRS-1. The PI3K inhibitors wortmannin and LY294002 inhibit the PDGF-induced phosphorylation of IRS-1, whereas the MEK inhibitor PD98059 was without a major effect. PDGF pretreatment for 60-90 min led to a marked 80-90% reduction in insulin stimulatable phosphotyrosine and IRS-1 associated PI3K activity. We examined the functional consequences of this decrease in IRS-1-associated PI3K activity. Interestingly, insulin stimulation of GLUT4 translocation and glucose transport was unaffected by 60-90 min of PDGF preincubation. Furthermore, insulin activation of Akt and p70(s6kinase), kinases downstream of PI3K, was unaffected by PDGF pretreatment. Wortmannin was capable of blocking these insulin actions following PDGF pretreatment, suggesting that PI3K was still necessary for these effects. In conclusion, 1) PDGF causes serine/threonine phosphorylation of IRS-1, and PI3K, or a kinase downstream of PI3K, mediates this phosphorylation. 2) This PDGF-induced phosphorylation of IRS 1 leads to a significant decrease in insulin-stimulated PI3K activity. 3) PDGF has no effect on insulin stimulation of Akt, p70(s6kinase), GLUT4 translocation, or glucose transport. 4) This suggests the existence of an IRS-1-independent pathway leading to the activation of PI3K, Akt, and p70(s6kinase); GLUT4 translocation; and glucose transport. PMID- 9737974 TI - Murine HIP/L29 is a heparin-binding protein with a restricted pattern of expression in adult tissues. AB - Heparin/heparan sulfate (Hp/HS)-binding proteins are implicated in a variety of cell biological processes including cell adhesion, modulation of blood coagulation, and cytokine/growth factor action. Hp/HS-interacting protein (HIP) has been identified in various adult tissues in humans. HIP supports high affinity, selective binding to Hp/HS, promotes cell adhesion, and modulates blood coagulation activities via Hp/HS-dependent mechanisms. Herein, a murine ortholog of human HIP is described that is 78.8% identical to human HIP and 99.8% identical at the cDNA level and identical at the amino acid level to a previously described murine ribosomal protein, L29. Western blot analyses and immunohistological staining with affinity-purified antibodies generated against two distinct peptide sequences of murine HIP/L29 indicate that HIP/L29 is differentially expressed in adult murine tissues and cell types. In the normal murine mammary epithelial cell line, NMuMG, HIP/L29 is enriched in the 100,000 x g particulate fraction. HIP/L29 can be solubilized from the 100,000 x g particulate fraction with 0.8 M NaCl, suggesting that it is a peripheral membrane protein. HIP/L29 directly binds 125I-Hp in gel overlay assays and requires 0.75 M NaCl for elution from Hp-agarose. In addition, recombinant murine HIP expressed in Escherichia coli binds Hp in a saturable and highly selective manner, compared with other glycosaminoglycans including dermatan sulfate, chondroitin sulfate, keratan sulfate, and hyaluronic acid. Collectively, these data indicate that murine HIP/L29, like its human ortholog, is a Hp-binding protein expressed in a restricted manner in adult tissues. PMID- 9737975 TI - Role of alpha-subunit of mitochondrial processing peptidase in substrate recognition. AB - Mitochondrial processing peptidase is a heterodimer consisting of alpha mitochondrial processing peptidase (alpha-MPP) and beta-MPP. We investigated the role of alpha-MPP in substrate recognition using a recombinant yeast MPP. Disruption of amino acid residues between 10 and 129 of the alpha-MPP did not essentially impair binding activity with beta-MPP and processing activity, whereas truncation of the C-terminal 41 amino acids led to a significant loss of binding and processing activity. Several acidic amino acids in the region conserved among the enzymes from various species were mutated to asparagine or glutamine, and effects on processing of the precursors were analyzed. Glu353 is required for processing of malate dehydrogenase, aspartate aminotransferase, and adrenodoxin precursors. Glu377 and Asp378 are needed only for the processing of aspartate aminotransferase and adrenodoxin precursors, both of which have a longer extension peptide than the others studied. However, processing of the yeast alpha-MPP precursor, which has a short extension peptide of nine amino acids, was not affected by these mutations. Thus, effects of substitution of acidic amino acids on the processing differed with the precursor protein and depended on length of the extension peptides. alpha-MPP may function as a substrate-recognizing subunit by interacting mainly with basic amino acids at a region distal to the cleavage site in precursors with a longer extension peptide. PMID- 9737976 TI - Regulation of mitogen-activated protein kinase phosphatase-1 induction by insulin in vascular smooth muscle cells. Evaluation of the role of the nitric oxide signaling pathway and potential defects in hypertension. AB - In this study, we examined the regulation of mitogen-activated protein kinase phosphatase (MKP-1) expression by insulin in primary vascular smooth muscle cell cultures. Insulin caused a rapid time- and dose-dependent induction of MKP-1 mRNA and protein expression. Blockade of nitric-oxide synthase (NOS) with NG monomethyl-L-arginine acetate, and cGMP with RpcGMP, completely inhibited MKP-1 expression. Insulin-mediated MKP-1 expression was preceded by inducible NOS (iNOS) induction and cGMP production. Blockade of phosphatidylinositol 3-kinase (PI3-kinase) signaling with wortmannin inhibited insulin-mediated iNOS protein induction, cGMP production, and MKP-1 expression. To evaluate potential interactions between NOS and the mitogen-activated protein kinase (MAPK) signaling pathways, we employed PD98059 and SB203580, two specific inhibitors of ERKs and p38 MAPK. These inhibitors abolished the effect of insulin on MKP-1 expression. Only PD98059 inhibited insulin-mediated iNOS protein induction. Vascular smooth muscle cells from spontaneous hypertensive rats exhibited a marked decrease in MKP-1 induction due to defects in insulin-induced iNOS expression because of reductions in PI3-kinase activity. Treatment with sodium nitroprusside and 8-bromo-cGMP restored MKP-1 mRNA expression to levels comparable with controls. We conclude that insulin-induced MKP-1 expression is mediated by PI3-kinase-initiated signals, leading to the induction of iNOS and elevated cGMP levels that stimulates MKP-1 expression. PMID- 9737977 TI - Identification of Nck family genes, chromosomal localization, expression, and signaling specificity. AB - Already a dozen molecules share binding to the Src homology (SH) 3 domains of human Nck, an SH3-SH3-SH3-SH2 adapter protein. We reason that there may be multiple gene members of Nck to accommodate the large binding repertoires. Here we report identification of novel human and mouse Nck genes and rename them as the Nckalpha and Nckbeta genes (including the human Nckalpha, human Nckbeta, mouse Nckalpha, and mouse Nckbeta genes). Nckalpha and Nckbeta share 68% amino acid identity, whereas the two Nckalpha and two Nckbeta across the species show 96% identity to each other. The human Nckbeta gene is mapped to 2q12, whereas the human Nckalpha gene has previously been mapped at 3q21. Antibodies specifically against Nckalpha and Nckbeta detect Nckalpha and Nckbeta with an identical molecular mass in the same cells of various origins. Ectopically expressed Nckbeta, but not its SH2 domain mutant, strongly inhibits epidermal growth factor and platelet-derived growth factor-stimulated DNA synthesis. Consistently, epidermal growth factor receptor and platelet-derived growth factor receptor preferentially interact with Nckbeta over Nckalpha in vitro. This study indicates that Nck is a multiple gene family and that each gene may have its own signaling specificity. Because previous anti-Nck (human Nckalpha) antibodies cross-react with Nckbeta, reassessment of those studies with specific Nck genes would be necessary. PMID- 9737978 TI - Evidence for heme-mediated redox regulation of human cystathionine beta-synthase activity. AB - Human cystathionine beta-synthase catalyzes the first step in the catabolic removal of the toxic metabolite, homocysteine. It is unique in being dependent on both pyridoxal phosphate (PLP) and heme for activity. The reaction involves condensation of serine and homocysteine to give cystathionine. Although the role of PLP can be rationalized in analogy with other PLP-dependent enzymes that catalyze beta-replacement reactions, the role of the heme is unknown. In this study, we have purified and characterized the recombinant human enzyme and have examined the effect of heme oxidation state on enzyme activity. We find that under reducing conditions, generated by addition of titanium citrate, the enzyme exhibits a 1.7-fold lower activity than under oxidizing conditions. Reoxidation of the ferrous enzyme with ferricyanide results in alleviation of inhibition. This redox-linked change in enzyme activity correlates with changes in heme oxidation state monitored by UV-visible spectroscopy. Dithiothreitol, which does not reduce the enzyme-bound heme, does not perturb enzyme activity. These studies provide the first evidence for redox-linked regulation of cystathionine beta synthase which is heme-dependent. PMID- 9737979 TI - IkappaB kinases serve as a target of CD28 signaling. AB - Optimal T cell activation and interleukin-2 production requires a second signal in addition to antigen-mediated T cell receptor (TCR) signaling. The CD28 molecule has been demonstrated to act as an effective costimulatory molecule upon binding by B7.1 or B7.2 present on antigen-presenting cells. The CD28 signal acts in concert with the TCR signal to significantly augment activation of the NF kappaB family of transcription factors. The interleukin-2 gene is regulated by NF kappaB among other transcription factors, in part, via a CD28 responsive element (CD28RE) present in the IL-2 promoter. Enhanced activation of NF-kappaB by CD28 is mediated by rapid phosphorylation and proteasome-mediated degradation of the NF-kappaB inhibitory proteins IkappaB alpha and IkappaB beta, which allows for accelerated nuclear expression of the liberated NF-kappaB. Herein, we provide evidence that the catalytic activities of two recently identified IkappaB kinases, IKKalpha and IKKbeta, are significantly elevated when T cells are stimulated through CD28 in addition to mitogen treatment. Catalytically inactive forms of IKKs are able to block the in vivo phosphorylation of IkappaB alpha induced by mitogen and CD28. Furthermore, CD28-mediated reporter gene transactivation of the CD28RE/AP-1 composite element is consistently attenuated by the IKK mutants. These findings suggest that cellular signaling pathways initiated at the TCR and CD28 converge at or upstream of IKK, resulting in more robust kinase activity and enhanced and prolonged NF-kappaB activation. PMID- 9737980 TI - Induction of angiotensin I-converting enzyme transcription by a protein kinase C dependent mechanism in human endothelial cells. AB - Angiotensin I-converting enzyme (ACE) has been implicated in various cardiovascular diseases; however, little is known about the ACE gene regulation in endothelial cells. We have investigated the effect of the protein kinase C activator phorbol 12-myristate 13-acetate (PMA) on ACE activity and gene expression in human umbilical vein endothelial cells (HUVEC). Our results showed a 3- and 5-fold increase in ACE activity in the medium and in the cells, respectively, after 24-h stimulation by PMA. We also observed an increase in the cellular ACE mRNA content starting after 6 h and reaching a 10-fold increase at 24 h in response to 100 ng/ml PMA as measured by ribonuclease protection assay. This effect was mediated by an increased transcription of the ACE gene as demonstrated by nuclear run-on experiments and nearly abolished by the specific PKC inhibitor GF 109203X. Our results indicate that PMA-activated PKC strongly increases ACE mRNA level and ACE gene transcription in HUVEC, an effect associated with an increased ACE secretion. A role for early growth response factor-1 (Egr-1) as a factor regulating ACE gene expression is suggested by both the presence of an Egr-1-responsive element in the proximal portion of the ACE promoter and the kinetics of the Egr-1 mRNA increase in HUVEC treated with PMA. PMID- 9737981 TI - Tumor suppressor p53 as a component of the tumor necrosis factor-induced, protein kinase PKR-mediated apoptotic pathway in human promonocytic U937 cells. AB - Despite what is known about the early signaling events in tumor necrosis factor (TNF) alpha-induced apoptosis, characterization of the downstream events remains largely undefined. It is now known that a cross-talk exists between the interferon and TNF-alpha pathways. This linkage allows recruitment of the cell proliferation suppressor PKR (dsRNA-dependent protein kinase) from the interferon pathway to play a pivotal role in TNF-alpha-induced apoptosis. In this study, we took advantage of the differential TNF-alpha susceptibilities of human promonocytic U937 subclones, deficient in or overexpressing PKR, to further characterize the role of PKR in apoptosis. By reverse transcription-polymerase chain reaction, we demonstrated that TNF-alpha transiently induces the tumor suppressor p53 in U937 cells. This p53 induction lags behind the TNF-alpha induction of PKR by 1 h. By cell viability determination, ultrastructural studies, apoptotic DNA laddering, and antisense techniques, it was shown that inhibition of p53 expression in PKR-overexpressing U937 cells abrogates the TNF alpha-induced apoptosis in these cells. Conversely, overexpressing wild type p53 in PKR-deficient U937 cells confers the susceptibility of these cells to TNF alpha-induced apoptosis. This latter result indicates that p53 induction is an event downstream of TNF-alpha-induced up-regulation of PKR, thereby further establishing the critical role of p53 in TNF-alpha-induced apoptosis in U937 cells. PKR-overexpressing U937 cells were found to possess a constitutively higher level of p53, which partly explains why these cells spontaneously undergo apoptosis even without TNF-alpha treatment. Finally, a model is presented on the interplay between PKR and p53 in effecting TNF-alpha-induced apoptosis in U937 cells. PMID- 9737982 TI - Testing the charge difference hypothesis for the assembly of a eucaryotic multispanning membrane protein. AB - The Glut1 glucose transporter is a glycoprotein whose membrane topology has been verified by a number of experimental observations, all of which are consistent with a 12-transmembrane helix model originally based on hydrophobicity analysis. We used Glut1 as a model multispanning membrane protein to test the Charge Difference Hypothesis (Hartmann, E., Rapoport, T. A., and Lodish, H. F. (1989) Proc. Natl. Acad. Sci. U. S. A. 86, 5786-5790), which asserts that the topology of a eucaryotic multispanning membrane protein is determined solely by the amino acid charge difference across the first transmembrane segment. The charge difference across the first transmembrane segment of Glut1 was progressively inverted in two independent series of mutants, one series in which only the number of positively charged amino acid residues in the two flanking domains was altered and the other in which only the number of negatively charged residues in the two flanking domains was changed. The results indicate that the charge difference across the first transmembrane segment does affect the topology of the protein, but that contrary to the hypothesis, it only dictates the orientation of the first transmembrane segment and the disposition of the amino terminus and the first linker domain. Charge inversion resulted in the formation of aberrant molecules in which either the first or second transmembrane segment failed to insert into the membrane. The topology of downstream regions of Glut1 was unaffected by charge inversion across the first transmembrane segment, indicating that downstream sequences are important in determining the local topological disposition of the molecule. PMID- 9737983 TI - A novel nuclear receptor heterodimerization pathway mediated by orphan receptors TR2 and TR4. AB - A unique heterodimerization pathway involving orphan receptors TR2 and TR4 is demonstrated. TR2 and TR4 preferentially form heterodimers in solution as well as on DNA elements containing a direct repeat-5 (DR5). The in vitro interaction between TR2 and TR4 is demonstrated by the yeast and the mammalian two-hybrid interaction assays, the pull-down assay, and the gel mobility shift assay. The in vivo interaction is demonstrated by following the intracellular localization of fusion receptors tagged with a green fluorescent protein. The dimerization is mediated by the ligand binding domains, and the three leucine residues on helix 10 of TR2 are critical for this interaction. In addition, coexpression of these two receptors exerts a much stronger repressive activity on a DR5-containing reporter than expressing either receptor alone. In the developing testis, TR2 and TR4 are coexpressed in the same testicular cell populations and exhibit a parallel pattern of expression along development. The preferential heterodimerization between TR2 and TR4 and their coexistence in specific germ cell populations suggest a physiological role of TR2/TR4 heterodimers in germ cell development. PMID- 9737984 TI - The N-end rule pathway catalyzes a major fraction of the protein degradation in skeletal muscle. AB - In skeletal muscle, overall protein degradation involves the ubiquitin-proteasome system. One property of a protein that leads to rapid ubiquitin-dependent degradation is the presence of a basic, acidic, or bulky hydrophobic residue at its N terminus. However, in normal cells, substrates for this N-end rule pathway, which involves ubiquitin carrier protein (E2) E214k and ubiquitin-protein ligase (E3) E3alpha, have remained unclear. Surprisingly, in soluble extracts of rabbit muscle, we found that competitive inhibitors of E3alpha markedly inhibited the 125I-ubiquitin conjugation and ATP-dependent degradation of endogenous proteins. These inhibitors appear to selectively inhibit E3alpha, since they blocked degradation of 125I-lysozyme, a model N-end rule substrate, but did not affect the degradation of proteins whose ubiquitination involved other E3s. The addition of several E2s or E3alpha to the muscle extracts stimulated overall proteolysis and ubiquitination, but only the stimulation by E3alpha or E214k was sensitive to these inhibitors. A similar general inhibition of ubiquitin conjugation to endogenous proteins was observed with a dominant negative inhibitor of E214k. Certain substrates of the N-end rule pathway are degraded after their tRNA dependent arginylation. We found that adding RNase A to muscle extracts reduced the ATP-dependent proteolysis of endogenous proteins, and supplying tRNA partially restored this process. Finally, although in muscle extracts the N-end rule pathway catalyzes most ubiquitin conjugation, it makes only a minor contribution to overall protein ubiquitination in HeLa cell extracts. PMID- 9737985 TI - Tyrosine 130 is an important outer ring donor for thyroxine formation in thyroglobulin. AB - The thyroid couples two iodotyrosine molecules to produce thyroid hormone at the acceptor site in thyroglobulin, leaving dehydroalanine or pyruvate at the donor position. Previous work has located the acceptors but not the principal iodotyrosine donors. We incorporated [14C]tyrosine into beef thyroid slices, isolated and iodinated the [14C]thyroglobulin (Tg I), separated its N-terminal approximately 22-kDa hormone-rich peptide, and digested the latter with trypsin and endoproteinase Glu-C (EC 3.4.21.19). Nonlabeled thyroglobulin (Tg II) was isolated from the same glands and processed similarly, without iodination in vitro. Tg I was used to initially recognize pyruvate in peptide fractions, and Tg II was used to then identify its location in the thyroglobulin polypeptide chain. Sequencing of a tryptic peptide by mass spectrometry and Edman degradation showed a cleavage after Val129. An endoproteinase Glu-C-generated peptide had the predicted molecular mass of a fragment containing residues 130-146 with Tyr130 replaced by pyruvate; the identification of this peptide was supported by obtaining the expected shortened fragment after tryptic digestion. 14C-labeled pyruvate was identified in the same fraction as this peptide. We conclude that Tyr130 is an important donor of the outer iodothyronine ring. Its likely acceptor is Tyr5, the most important hormonogenic site of thyroglobulin, because Tyr5 and Tyr130 are proximate, because they are the most prominent early iodination sites in this part of thyroglobulin, and because the N-terminal region was previously found capable of forming T4 by itself. PMID- 9737986 TI - Conserved cysteines in the type 1 deiodinase selenoprotein are not essential for catalytic activity. AB - The iodothyronine deiodinases are a family of oxidoreductases that catalyze the removal of iodide from thyroid hormones. Each of the three isoforms contain selenocysteine at its active site and several cysteine residues that may be important for catalytic activity. Of particular interest in the type I deiodinase (D1) is Cys124, which is vicinal to the selenocysteine at position 126, and Cys194, which has been conserved in all deiodinases identified to date. In the present studies, we have characterized the functional properties of C124A, C194A, and C124A/C194A D1 mutants, which were prepared by site-directed mutagenesis and expressed in COS-7 cells. In broken cell preparations, the sensitivity of the mutants to the selective D1 inhibitors propylthiouracil and aurothioglucose were unaltered. Mutagenesis at the Cys124 position was associated with a 7-11-fold increase in the Km of dithiothreitol, whereas Vmax values remained largely unchanged. However, both mutations resulted in marked decreases in Vmax values when glutathione or a reconstituted thioredoxin cofactor system were used in the assay. In contrast to the results of these in vitro studies, no impairment in deiodinating capability was noted in intact cells expressing equivalent levels of the mutant constructs. These studies demonstrate that Cys124 and Cys194 influence the reactivity of the D1 with thiol cofactors in in vitro assay systems but are not determinants of the sensitivity of the enzyme to propylthiouracil and aurothioglucose. Furthermore, the observation that the cysteine mutants are fully active in intact cells demonstrates that the results of commonly used broken cell assays do not accurately predict the activity of the D1 in intact cells and suggests that glutathione and thioredoxin are not the major thiols utilized in vivo to support D1 activity. PMID- 9737987 TI - Characterization of the aldolase B intronic enhancer. AB - The aldolase B gene is transcribed at a high level in the liver, kidney, and small intestine. This high level of gene expression results from cooperation between a weak but liver-specific promoter and an intronic activator. A deletional study of this activator present in the first intron allowed us to ascribe the maximal enhancer function to a 400-base pair (bp) fragment (+1916 to + 2329). This enhancer is highly liver-specific and enhances the activity of heterologous minimal promoters in a position and distance-independent fashion in transiently transfected Hep G2 hepatoma cells. The aldolase B enhancer is composed of two domains, a 200-bp module (Ba) inactive by itself but which synergizes with another 200-bp module (Bb) that alone retains 25% of the total enhancer activity. The Bb sequence is 76% homologous between human and rat genes and contains several binding sites for liver-enriched nuclear factors. By electrophoretic mobility shift assays, we demonstrated that elements 5 and 7 bind hepatic nuclear factor 1 (HNF1), whereas element 2 binds hepatic nuclear factor 4 (HNF4). A functional analysis of the enhancer whose elements have been mutated demonstrated that mutation of any of the HNF1 sites totally suppressed enhancer activity, whereas mutation of the HNF4-binding site reduced it by 80%. PMID- 9737988 TI - Human alpha1,3/4-fucosyltransferases. I. Identification of amino acids involved in acceptor substrate binding by site-directed mutagenesis. AB - In a previous study (Xu, Z., Vo, L., and Macher, B. A. (1996) J. Biol. Chem. 271, 8818-8823), a domain swapping approach demonstrated that a region of amino acids found in human alpha1, 3/4-fucosyltransferase III (FucT III) conferred a significant increase in alpha1,4-FucT acceptor substrate specificity into alpha1, 3-fucosyltransferase V (FucT V), which, under the same assay conditions, has extremely low alpha1,4-FucT acceptor substrate specificity. In the current study, site-directed mutagenesis was utilized to identify which of the eight amino acids, associated with alpha1,4-FucT acceptor substrate specificity, is/are responsible for conferring this new property. The results demonstrate that increased alpha1,4-FucT activity with both disaccharide and glycolipid acceptors can be conferred on FucT V by modifying as few as two (Asn86 to His and Thr87 to Ile) of the eight amino acids originally swapped from FucT III into the FucT V sequence. Neither single amino acid mutant had increased alpha1,4-FucT activity relative to that of FucT V. Kinetic analyses of FucT V mutants demonstrated a reduced Km for Galbeta1,3GlcNAc (type 1) acceptor substrates compared with native FucT V. However, this was about 20-fold higher than that found for native FucT III, suggesting that other amino acids in FucT III must contribute to its overall binding site for type 1 substrates. These results demonstrate that amino acid residues near the amino terminus of the catalytic domain of FucT III contribute to its acceptor substrate specificity. PMID- 9737989 TI - Human alpha1,3/4-fucosyltransferases. II. A single amino acid at the COOH terminus of FucT III and V alters their kinetic properties. AB - An analysis of the acceptor substrate specificity of domain swap mutants of human alpha1,3/4-fucosyltransferases (FucTs) III and V has been carried out. The results demonstrate that changing Asp336 of FucT III to Ala (as in FucT V) produced a protein (III/V1) with a reduced activity with a variety of acceptors. An analysis of the kinetic properties of FucT III and the III/V1 mutant demonstrated that III/V1 had a 40-fold reduction in its affinity for the H-type 1 acceptor substrate (Fucalpha1,2Galbeta1,3GlcNAc) and 4-fold reduction in its affinity for GDP-fucose when compared with FucT III. Further, the overall catalytic efficiency of III/V1 was approximately 100-fold lower than that of FucT III with an H-type 1 acceptor substrate. The complementary domain swap resulting from the change of Ala349 of FucT V to Asp (V/III1) produced a FucT that had higher enzyme activity with a range of acceptor substrates and had a higher affinity for an H-type 2 acceptor substrate (Fucalpha1, 2Galbeta1,4GlcNAc) with an 8-fold higher overall catalytic efficiency than that of FucT V. No significant change occurred in the Km for GDP-fucose for this protein when compared with FucT V. Kinetic parameters of two other FucT domain swaps (III8/V and V8/III), resulting in proteins that differed from FucT III and V at the NH2 terminus of their catalytic domain, were not significantly different from those of the parental enzymes when H-type 1 and H-type 2 acceptor substrates were utilized. Thus, substitution of an acidic amino acid for a nonpolar amino acid (i.e. Asp versus Ala) at the COOH terminus of FucTs produces an enzyme with enhanced enzyme activities. These results, together with the results presented in the accompanying papers (Nguyen, A. T., Holmes, E. H., Whitaker, J. M., Ho, S., Shetterly, S., and Macher, B. A. (1998) J. Biol. Chem. 273, 25244-25249; Sherwood, A. L., Nguyen, A. T., Whitaker, J. M., Macher, B. A., and Holmes, E. H. (1998) J. Biol. Chem. 273, 25256-25260), provide new insights into the structure/function relationships of human alpha1,3/4-FucT enzymes. PMID- 9737990 TI - Human alpha1,3/4-fucosyltransferases. III. A Lys/Arg residue located within the alpha1,3-FucT motif is required for activity but not substrate binding. AB - Amino acid sequence alignment of human alpha1, 3/4-fucosyltransferases (FucTs) demonstrates that three highly conserved Lys residues are present in the catalytic domain of FucTs III, IV, V, and VI. Two of these sites are conserved in FucT VII, with the third located within the alpha1,3-FucT motif as a conservative change to Arg at position 223. Site-directed mutagenesis experiments were conducted to change Lys255 of FucT V (equivalent to Arg223 of FucT VII) to either Arg255 or Ala255. Enzyme assays demonstrate that the FucT V K255R mutant has a 34 fold lower specific activity than native FucT V and that the K255A mutant is inactive. Site-directed mutagenesis of FucT VII was also conducted to change Arg223 to Lys223 for analysis of the effect on enzyme kinetic parameters. No differences in acceptor specificities or Km values for either substrate were observed between native FucT VII and the R223K mutant; however, the purified R223K mutant enzyme had a 2-fold increased specific activity compared with purified native FucT VII. No change in GDP-fucose-protectable pyridoxal-P/NaBH4 inactivation was observed for native or mutant FucT V or VII, further supporting the absence of involvement of this residue in sugar nucleotide binding. The results indicate that a basic residue in this position is required for enzyme activity, with a Lys residue providing higher intrinsic activity. The lack of influence of this site on substrate binding parameters and its location within the alpha1,3-FucT motif suggest that at least some of the residues within this motif are involved in catalysis rather than substrate binding. PMID- 9737991 TI - Purification and characterization of a polysome-associated endoribonuclease that degrades c-myc mRNA in vitro. AB - The regulation of mRNA half-lives is determined by multiple factors, including the activity of the messenger RNases (mRNases) responsible for destroying mRNA molecules. Previously, we used cell-free mRNA decay assays to identify a polysome associated endonuclease that cleaves c-myc mRNA within the coding region. A similar activity has been solubilized and partially purified from a high salt extract of adult rat liver polysomes. Based on a correlation between protein and enzyme activity, the endonuclease is tentatively identified as a approximately 39 kDa protein. It cleaves the coding region stability determinant of c-myc mRNA with considerable specificity. Cleavages occur predominantly in an A-rich segment of the RNA. The endonuclease is resistant to RNase A inhibitors, sensitive to vanadyl ribonucleoside complex, and dependent on magnesium. In these and other respects, the soluble enzyme we have purified resembles the polysome-associated c myc mRNase. PMID- 9737992 TI - Oxidation-reduction properties of methylglyoxal-modified protein in relation to free radical generation. AB - Oxidation-reduction properties of methylglyoxal-modified protein in relation to free radical generation were investigated. Glycation of bovine serum albumin by methylglyoxal generated the protein-bound free radical, probably the cation radical of the cross-linked Schiff base, as observed in the reaction of methylglyoxal with L-alanine (Yim, H.-S., Kang, S.-O., Hah, Y. C., Chock, P. B., and Yim, M. B. (1995) J. Biol. Chem. 270, 28228-28233) or with Nalpha-acetyl-L lysine. The glycated bovine serum albumin showed increased electrophoretic mobility suggesting that the basic residues, such as lysine, were modified by methylglyoxal. The glycated protein reduced ferricytochrome c to ferrocytochrome c in the absence of oxygen or added metal ions. This reduction of cytochrome c was accompanied by a large increase in the amplitude of the electron paramagnetic resonance signal originated from the protein-bound free radical. In addition, the glycated protein catalyzed the oxidation of ascorbate in the presence of oxygen, whereas the protein free radical signal disappeared. These results indicate that glycation of protein generates active centers for catalyzing one-electron oxidation-reduction reactions. This active center, which exhibits enzyme-like characteristic, was suggested to be the cross-linked Schiff base/the cross-linked Schiff base radical cation of the protein. It mimics the characteristics of the metal-catalyzed oxidation system. The glycated bovine serum albumin cross-linked further to the cytochrome c in the absence of methylglyoxal. The cross-linked cytochrome c maintains its oxidation-reduction properties. These results together indicate that glycated proteins accumulated in vivo provide stable active sites for catalyzing the formation of free radicals. PMID- 9737993 TI - Antibody cross-linking of the glycosylphosphatidylinositol-linked protein CD59 on hematopoietic cells induces signaling pathways resembling activation by complement. AB - CD59 is a glycosylphosphatidylinositol-anchored cell surface glycoprotein involved in protecting cells from host-mediated complement attack. Studies have shown that antibody cross-linking of CD59 induces a series of intracellular signaling events including the activation of protein-tyrosine kinases (PTK). To further characterize these events, antibodies and complement 8, one of the natural ligands of CD59, were used to activate CD59. Antibody-induced cross linking of CD59 on the surface of THP-1 and U937 hematopoietic cell lines as well as exposure to complement 8 induces a rapid increase in the tyrosine phosphorylation of several proteins within the cell. Consistent with an early role for the Src family PTKs in these signaling events, we found that transient activation of Hck- and CD59-mediated signaling was abrogated in the presence of the Src family PTK-selective inhibitor PP1. Although the molecular mechanism by which CD59 communicates to Hck is unknown, cellular fractionation studies indicated that both CD59 and Hck are compartmentalized in plasma membrane microdomains. We also detected tyrosine phosphorylation of the adaptor proteins p120 and Shc, and the cytoplasmic nonreceptor tyrosine kinase Syk. The identification of CD59-mediated signaling events may help explain why paroxysmal nocturnal hemoglobinuria patients, who are deficient in glycosylphosphatidylinositol-linked proteins including CD59, are susceptible to proliferative disorders. PMID- 9737994 TI - HERG potassium channel activation is shifted by phorbol esters via protein kinase A-dependent pathways. AB - We investigated the effects of the phorbol ester phorbol 12-myristate 13-acetate (PMA) on the rapid component of the delayed rectifier potassium current, IKr, in guinea pig cardiomyocytes and found that the IKr current amplitude was reduced by 20% with 10 nM PMA and 44% with 100 nM PMA. The ether-a-go-go-related gene (HERG) encodes IKr in human heart. We expressed HERG heterologously in Xenopus oocytes and investigated the effects of PMA on the delayed rectifier potassium current. Upon application of PMA in a concentration of 100 nM, we found a similar reduction of HERG outward current amplitude by 59%. This reduction was due to a shift in the HERG activation curve by 37 mV. The ED50 for the PMA-induced shift was 9.0 nM. The inactive 4alpha-phorbol 12-myristate 13-acetate (4alpha-PMA) had no effect. PMA is known to act by stimulating distinct protein kinase cascades. Additional application of the specific protein kinase C inhibitors chelerythrine (10 microM) or bisindolylmaleimide (1 microM) could not attenuate the PMA-induced shift. In contrast, the shift by PMA was reduced significantly when the specific protein kinase A (PKA) inhibitors H89 (50 microM) or KT5720 (2.5 microM) were applied. Forskolin (400 microM), an activator of the adenylate cyclase that results in PKA activation, shifted the HERG activation curve by 14 mV. Moreover the specific protein kinase C activator 1-stearoyl-2-arachidonylglycerol (10 microM) showed no effect. Our data suggest that mainly PKA is mediating the shift of the HERG activation kinetics. PMID- 9737995 TI - A novel distal enhancer confers chorionic expression on the human renin gene. AB - Renin catalyzes the rate-limiting step of the renin-angiotensin system, which regulates blood pressure and electrolyte homeostasis. To determine cell-specific human renin gene control elements, the transcriptional activity of promoter regions up to position -8876 was studied in renin-expressing cells. A positive regulatory region conferring approximately 57-fold higher transcriptional activity to the human renin gene promoter in chorionic cells was identified between nucleotides -5777 and -5552. It had the orientation-independent activity typical of classical enhancers. It also conferred approximately 59-fold higher transcriptional levels from the heterologous simian virus 40 (SV40) promoter in chorionic cells and approximately 6-fold higher transcriptional levels in Calu-6 and As4.1 cells, whereas no effect was measured in non-renin-expressing cells. DNase I footprinting showed that this enhancer contains three binding sites for chorionic cell nuclear extracts. Functional analysis suggested that the activity of the enhancer is regulated by differential mechanisms in the three renin expressing cells involving a complex arrangement of AP-1 motifs binding cell specific members of the basic leucine zipper family of transcription factors. Thus, our results demonstrate that this enhancer plays a key role in the expression of the human renin gene in the chorion and may also be involved in its regulated expression in other tissues. PMID- 9737996 TI - Inhibition of in vitro endosomal vesicle fusion activity by aminoglycoside antibiotics. AB - The effects of two aminoglycoside antibiotics, neomycin and Geneticin, on the endocytic pathway were studied using a cell-free assay that reconstitutes endosome-endosome fusion. Both drugs inhibit the rate and extent of endosome fusion in a dose-dependent manner with IC50 values of approximately 45 microM and approximately 1 mM, respectively. Because the IC50 for neomycin falls within the range of affinities reported for its binding to acidic phospholipids, notably phosphatidylinositol 4,5-bisphosphate (PIP2), these data suggest that negatively charged lipids are required for endosome fusion. A role for negatively charged lipids in membrane traffic has been postulated to involve the activity of a PIP2 dependent phospholipase D (PLD) stimulated by the GTP-binding protein ADP ribosylation factor (ARF). Although neomycin blocks endosome fusion at a stage of the in vitro reaction that is temporally related to steps inhibited by cytosolic ARFs when they bind guanosine-5'-gamma-thiophosphate (GTPgammaS), these inhibitors appear to act in a synergistic manner. This idea is confirmed by the fact that addition of a PIP2-independent PLD does not suppress neomycin inhibition of endosome fusion; moreover, in vitro fusion activity is not affected by the pleckstrin homology domain of phosphoinositide-specific phospholipase C delta1, which binds to acidic phospholipids, particularly PIP2, with high affinity. Thus, although aminoglycoside-sensitive elements of endosome fusion are required at mechanistic stages that are also blocked by GTPgammaS-bound ARF, these effects are unrelated to inhibition of the PIP2-dependent PLD activity stimulated by this GTP-binding protein. These results argue that there are additional mechanistic roles for acidic phospholipids in the endosomal pathway. PMID- 9737997 TI - Forced expression of Id-1 in the adult mouse small intestinal epithelium is associated with development of adenomas. AB - Ids are dominant-negative helix-loop-helix (HLH) proteins that play overlapping yet distinct roles in antagonizing basic HLH transcription factors. Although Ids affect myogenesis, neurogenesis, and B-cell development, little is known about their in vivo functions in epithelia. We have examined the effects of forced expression of Id-1 in the small intestinal epithelium of adult chimeric mice. 129/Sv embryonic stem cells, transfected with DNA containing Id-1 under the control of transcriptional regulatory elements that function in all intestinal epithelial cell lineages, were introduced into C57Bl/6 (B6) blastocysts heterozygous for the ROSA26 marker. The B6 ROSA26/+ intestinal epithelium of the resulting adult chimeras produces Escherichia coli beta-galactosidase, allowing identification of this internal control cell population. Chimeras produced from nontransfected embryonic stem cells served as additional controls. Immunohistochemical studies of the control chimeras indicated that the small intestinal epithelium supports a complex pattern of endogenous Id expression. Id 1 is restricted to the cytoplasm; levels do not decrease as descendants of multipotent intestinal stem cells differentiate. Id-2 and Id-3 are only detectable in nuclei; levels increase markedly as epithelial cells differentiate. Forced expression of Id-1 in the 129/Sv epithelium results in a decline in Id-2 and Id-3 to below the limits of immunodetection. A subset of chimeric-transgenic mice lacked growth factor- and defensin-producing Paneth cells in their 129/Sv epithelium and also developed intestinal adenomas. These changes were not present in normal control chimeras. Adenomas were composed of proliferating beta-Gal positive and -negative epithelial cells, suggesting that they arose through cooperative interactions between 129/Sv(Id-1) and B6 ROSA26/+ cells. These chimeras provide a model for studying how perturbations in Id expression affect tumorigenesis. PMID- 9737999 TI - Characterization of the structure, function, and conformational stability of PorB class 3 protein from Neisseria meningitidis. A porin with unusual physicochemical properties. AB - PorB proteins constitute the vast majority of channels in neisserial outer membranes and can be subdivided within meningococcal strains into two distinct and mutually exclusive families that are designated as class 2 and class 3 proteins. We recently characterized the functional activity and conformational stability of a PorB class 2 protein from Neisseria meningitidis (Minetti, C. A. S. A., Tai, J. Y., Blake, M. S., Pullen, J. K., Liang, S. M., and Remeta, D. P. (1997) J. Biol. Chem. 272, 10710-10720). To evaluate the structure-function relatedness among the PorB proteins, we have employed a combination of electrophoretic and spectroscopic techniques to assess the conformational stability of zwittergent-solubilized class 3 trimers. The functional, physicochemical, and structural properties of the meningococcal class 2 and class 3 proteins are comparable with the notable exception that the latter exhibits a significantly higher susceptibility to SDS. The SDS-induced dissociation and partial unfolding of PorB class 3 is characterized by a single two-state transition with a midpoint at 0.35% SDS. The native trimeric assembly dissociates reversibly, forming partially folded monomers that retain the characteristic beta sheet content of the transmembrane domain with a concomitant increase in random coil structure arising from unfolding the rigid surface loops. These results provide new insight into the elucidation of porin folding pathways and the factors that govern the overall structural stability of meningococcal proteins. PMID- 9737998 TI - Activation and oligomerization of aspartylglucosaminidase. AB - Secretory, membrane, and lysosomal proteins undergo covalent modifications and acquire their secondary and tertiary structure in the lumen of the endoplasmic reticulum (ER). In order to pass the ER quality control system and become transported to their final destinations, many of them are also assembled into oligomers. We have recently determined the three-dimensional structure of lysosomal aspartylglucosaminidase (AGA), which belongs to a newly discovered family of homologous amidohydrolases, the N-terminal nucleophile hydrolases. Members of this protein family are activated from an inactive precursor molecule by an autocatalytic proteolytic processing event whose exact mechanism has not been thoroughly determined. Here we have characterized in more detail the initial events in the ER required for the formation of active AGA enzyme using transient expression of polypeptides carrying targeted amino acid substitutions. We show that His124 at an interface between two heterodimers of AGA is crucial for the thermodynamically stable oligomeric structure of AGA. Furthermore, the side chain of Thr206 is essential both for the proteolytic activation and enzymatic activity of AGA. Finally, the proper geometry of the residues His204-Asp205 seems to be crucial for the activation of AGA precursor polypeptides. We propose here a reaction mechanism for the activation of AGA which could be valid for homologous enzymes as well. PMID- 9738000 TI - PC5-A-mediated processing of pro-neurotensin in early compartments of the regulated secretory pathway of PC5-transfected PC12 cells. AB - Among the members of the proprotein convertase (PC) family, PC1 and PC2 have well established roles as prohormone convertases. Another good candidate for this role is PC5-A that has been shown to be present in the regulated secretory pathway of certain neuroendocrine tissues, but evidence that it can process prohormones is lacking. To determine whether PC5-A could function as a prohormone convertase and to compare its cleavage specificity with that of PC1 and PC2, we stably transfected the rat pheochromocytoma PC12 cell line with PC5-A and analyzed the biosynthesis and subcellular localization of the enzyme, as well as its ability to process pro-neurotensin/neuromedin N (pro-NT/NN) into active peptides. Our data showed that in transfected PC12 cells, PC5-A was converted from its 126-kDa precursor form into a 117-kDa mature form and, to a lesser extent, into a C terminally truncated 65-kDa form of the 117-kDa product. Metabolic and immunochemical studies showed that PC5-A was sorted to early compartments of the regulated secretory pathway where it colocalized with immunoreactive NT. Furthermore, pro-NT/NN was processed in these compartments according to a pattern that differed from that previously described in PC1- and PC2-transfected PC12 cells. This pattern resembled that previously reported for pro-NT/NN processing in the adrenal medulla, a tissue known to express high levels of PC5-A. Altogether, these data demonstrate for the first time the ability of PC5-A to function as a prohormone convertase in the regulated secretory pathway and suggest a role for this enzyme in the physiological processing of pro-NT/NN. PMID- 9738002 TI - Association of the TLX-2 homeodomain and 14-3-3eta signaling proteins. AB - Homeodomain proteins play important roles in various developmental processes, and their functions are modulated by polypeptide cofactors. Here we report that both in vitro and in vivo, 14-3-3eta is associated with the TLX-2 homeodomain transcription factor that is required for mouse embryogenesis. Expression of 14-3 3eta shifts the predominant localization of TLX-2 in COS cells from the cytoplasm to the nucleus. Tlx-2 and 14-3-3eta are expressed in the developing peripheral nervous system with spatially and temporally overlapping patterns, and they are also coexpressed in PC12 cells. Increased expression of either gene by transfection considerably inhibited nerve growth factor-induced neurite outgrowth of PC12 cells, and cotransfection of both genes led to a synergistic effect of suppression. These findings define 14-3-3eta as a functional modulator of the TLX 2 homeodomain transcription factor and suggest that the in vivo function of TLX-2 in neural differentiation is likely regulated by signaling mediated by 14-3-3eta. PMID- 9738001 TI - Reconstitution of insulin signaling pathways in rat 3Y1 cells lacking insulin receptor and insulin receptor substrate-1. Evidence that activation of Akt is insufficient for insulin-stimulated glycogen synthesis or glucose uptake in rat 3Y1 cells. AB - Rat 3Y1 cells have endogenous insulin-like growth factor-1 receptors and insulin receptor substrate (IRS)-2, but lack both insulin receptor (IR) and IRS-1. To investigate the role of IR and IRS-1 in effects of insulin, we transfected IR and IRS-1 expression plasmids into cells and reconstituted the insulin signaling pathways. 3Y1 cells stably expressing the c-myc epitope-tagged glucose transporter type 4 (3Y1-GLUT4myc) exhibit no effects of insulin, at physiological concentrations. The 3Y1-GLUT4myc-IR cells expressing GLUT4myc and IR responded to phosphatidylinositol 3,4, 5-trisphosphate (PI-3,4,5-P3) accumulation, Akt activation, the stimulation of DNA synthesis, and membrane ruffling but not to glycogen synthesis, glucose uptake, or GLUT4myc translocation. The further expression of IRS-1 in 3Y1-GLUT4myc-IR cells led to stimulation of glycogen synthesis but not to glucose uptake or GLUT4myc translocation in response to insulin, although NaF or phorbol 12-myristate 13-acetate did trigger GLUT4myc translocation in the cells. These results suggest that, in rat 3Y1 cells, (i) IRS 1 is essential for insulin-stimulated glycogen synthesis but not for DNA synthesis, PI-3,4,5-P3 accumulation, Akt phosphorylation, or membrane ruffling, and (ii) the accumulation of PI-3,4,5-P3 and activation of Akt are insufficient for glycogen synthesis, glucose uptake or for GLUT4 translocation. PMID- 9738003 TI - Physical and functional interaction of murine and Xenopus Smad7 with bone morphogenetic protein receptors and transforming growth factor-beta receptors. AB - Members of the transforming growth factor-beta (TGF-beta) family transmit signals from membrane to nucleus via intracellular proteins known as Smads. A subclass of Smad proteins has recently been identified that antagonize, rather than transduce, TGF-beta family signals. Smad7, for example, binds to and inhibits signaling downstream of TGF-beta receptors. Here we report that the C-terminal MAD homology domain of murine Smad7 (mSmad7) is sufficient for both of these activities. In addition, we show that mSmad7 interacts with activated bone morphogenetic protein (BMP) type I receptors (BMPR-Is), inhibits BMPR-I-mediated Smad phosphorylation, and phenocopies the effect of known BMP antagonists when overexpressed in ventral cells of Xenopus embryos. Xenopus Smad7 (XSmad7, previously termed Smad8) and mSmad7 are nearly identical within their bioactive C domain, but have quite distinct N-domains. We found that XSmad7, similar to mSmad7, interacted with BMP and TGF-beta type I receptors and inhibited receptor mediated phosphorylation of downstream signal-transducing Smads. However, XSmad7 is a less efficient inhibitor of TbetaR-I-mediated responses in mammalian cells than is mSmad7. Furthermore, overexpression of XSmad7 in Xenopus embryos produces patterning defects that are not observed following overexpression of mSmad7, suggesting that mSmad7 and XSmad7 may preferentially target distinct signaling pathways. Our results are consistent with the possibility that the C-domain of antagonistic Smads is an effector domain whereas the N-domain may confer specificity for distinct signaling pathways. PMID- 9738004 TI - Combinatorial cis-acting elements control tissue-specific activation of the cardiac troponin I gene in vitro and in vivo. AB - The cardiac troponin I gene is one of the few sarcomeric protein genes exclusively expressed in cardiac muscle. We show here that this specificity is controlled by a proximal promoter (-230/+16) in transfected cardiac cells in culture, in the adult hearts, and in transgenic animals. Functional analysis indicates that MEF2/Oct-1, Sp1, and GATA regulatory elements are required for optimal gene activation because selective mutations produce weak or inactive promoters. MEF2 and Oct-1 transcription factors bind to the same A/T-rich element. A mutation that blocks this binding markedly reduces gene activation in vivo and in vitro, and overexpression of MEF2A, MEF2C, and MEF2D in noncardiac cells transactivates the cardiac troponin I promoter. Disruption of these elements inactivates the cardiac troponin I promoter in cultured cardiac cells but has a less important role in transfected adult heart. Moreover, nuclear extracts from an almost pure population of adult cardiac cells contain much lower levels of GATA binding activity compared with fetal cardiac cells. These findings point to a differential role of GATA factors in the maintenance of gene expression in the adult heart as compared with the activation of cardiac genes in fetal cardiomyocytes. Overexpression of GATA family members transactivates the cardiac troponin I promoter, and GATA-5 and GATA-6 are stronger transactivators than GATA-4, a property apparently unique to the cardiac troponin I promoter. Transgenic mice carrying the -230/+126 base pair promoter express beta galactosidase reporter gene in the heart both at early stages of cardiogenesis and in the adult animals. These results indicate that the ability of the cardiac troponin I proximal promoter to target expression of a downstream gene in the heart is also maintained when the transgene is integrated into the genome. PMID- 9738005 TI - Estrogen-dependent production of erythropoietin in uterus and its implication in uterine angiogenesis. AB - Although erythropoietin (Epo) has been shown to possess in vitro angiogenic activity, its physiological significance has not been demonstrated. Normally angiogenesis does not occur actively in adults but an exception is the female reproductive organ. In the uterine endometrium, angiogenesis takes place actively for supporting the endometrial growth that occurs during transition from the diestrus to estrous stage. This transition is under control of 17beta-estradiol (E2), an ovarian hormone, and can be mimicked by injection of E2 to ovariectomized (OVX) mouse. Thus, the uterus is a pertinent site to examine the Epo function in angiogenesis. We found that Epo protein and its mRNA were produced in an E2-dependent manner, when the uterus from OVX mouse was cultured in vitro. The de novo protein synthesis was not needed for E2 induction of Epo mRNA. Administration of E2 to OVX mouse induced a rapid and transient increase in Epo mRNA in the uterus. Injection of Epo into the OVX mouse uterine cavity promoted blood vessel formation in the endometrium. Furthermore, injection of the soluble Epo receptor capable of binding with Epo into the uterine cavity of non OVX mouse in diestrus stage inhibited the endometrial transition to proestrus stage, whereas heat-inactivated soluble Epo receptor allowed the transition to occur. These results, combined with our finding that the endothelial cells in uterine endometrium express Epo receptor, strongly suggest that Epo is an important factor for the E2-dependent cyclical angiogenesis in uterus. PMID- 9738006 TI - The C-terminal (BRCT) domains of BRCA1 interact in vivo with CtIP, a protein implicated in the CtBP pathway of transcriptional repression. AB - The BRCA1 tumor suppressor encodes a polypeptide with two recognizable protein motifs: a RING domain near the N terminus and two tandem BRCT domains at the C terminus. Studies of tumor-associated mutations indicate that the RING and BRCT sequences are required for BRCA1-mediated tumor suppression. In addition, recent work has shown that BRCA1 is a potent regulator of RNA transcription and that the BRCT domains are also essential for this activity. Therefore, we used the Sos recruitment system to screen for proteins that bind this critical region of BRCA1. Our results show that the BRCT domains interact in vivo with CtIP, a protein originally identified on the basis of its association with the CtBP transcriptional co-repressor. This finding suggests that BRCA1 regulates gene expression, at least in part, by modulating CtBP-mediated transcriptional repression. Moreover, the in vivo interaction between BRCA1 and CtIP is completely ablated by each of three independent tumor-associated mutations affecting the BRCT motifs of BRCA1. These results indicate that the BRCA1-CtIP interaction may be required for tumor suppression by BRCA1. PMID- 9738007 TI - Delineation of the functional site of alpha-dendrotoxin. The functional topographies of dendrotoxins are different but share a conserved core with those of other Kv1 potassium channel-blocking toxins. AB - We identified the residues that are important for the binding of alpha dendrotoxin (alphaDTX) to Kv1 potassium channels on rat brain synaptosomal membranes, using a mutational approach based on site-directed mutagenesis and chemical synthesis. Twenty-six of its 59 residues were individually substituted by alanine. Substitutions of Lys5 and Leu9 decreased affinity more than 1000 fold, and substitutions of Arg3, Arg4, Leu6, and Ile8 by 5-30-fold. Substitution of Lys5 by norleucine or ornithine also greatly altered the binding properties of alphaDTX. All of these analogs displayed similar circular dichroism spectra as compared with the wild-type alphaDTX, indicating that none of these substitutions affect the overall conformation of the toxin. Substitutions of Ser38 and Arg46 also reduced the affinity of the toxin but, in addition, modified its dichroic properties, suggesting that these two residues play a structural role. The other residues were excluded from the recognition site because their substitutions caused no significant affinity change. Thus, the functional site of alphaDTX includes six major binding residues, all located in its N-terminal region, with Lys5 and Leu9 being the most important. Comparison of the functional site of alphaDTX with that of DTX-K, another dendrotoxin (Smith, L. A., Reid, P. F., Wang, F. C., Parcej, D. N., Schmidt, J. J., Olson, M. A., and Dolly, J. O. (1997) Biochemistry 36, 7690-7696), reveals that they only share the predominant lysine and probably a leucine residue; the additional functional residues differ from one toxin to the other. Comparison of the functional site of alphaDTX with those of structurally unrelated potassium channel-blocking toxins from venomous invertebrates revealed the common presence of a protruding key lysine with a close important hydrophobic residue (Leu, Tyr, or Phe) and few additional residues. Therefore, irrespective of their phylogenetic origin, all of these toxins may have undergone a functional convergence. The functional site of alphaDTX is topographically unrelated to the "antiprotease site" of the structurally analogous bovine pancreatic trypsin inhibitor. PMID- 9738009 TI - Mode of binding of anti-P-glycoprotein antibody MRK-16 to its antigen. A crystallographic and molecular modeling study. AB - Monoclonal antibody MRK-16 recognizes a discontinuous extracellular epitope on the multidrug resistance-associated ATP-binding cassette transporter, P glycoprotein. The atomic basis for specificity of this antibody is of interest because of its potential as a modulator of P-glycoprotein activity. The crystal structure of Fab MRK-16 is reported to a resolution of 2.8 A. A structure for a portion of the epitope was derived by comparison to regions of solved structures with similar primary sequence. This has permitted a proposal for the mode of binding of the peptide epitope to the antibody, in which the peptide makes specific contacts with complementarity-determining regions H1, H2, and H3 from the heavy chain and L3 from the light chain. These interactions are consistent with epitope mapping studies and with the observation that MRK-16 is specific for human class I P-glycoprotein. This result identifies side chains in MRK-16 that would be amenable to alteration in antibody engineering experiments to derive improved multidrug resistance inhibitors for clinical use during chemotherapy. In particular, Arg-H97 contacts both Glu-746 and Asp-744 of the peptide, Arg-L96 contacts Asp-743, and Thr-H33 interacts with Thr-747. All of these epitope residues were implicated in mediating specificity by epitope mapping studies. PMID- 9738008 TI - Collagen XVIII is a basement membrane heparan sulfate proteoglycan. AB - The present study shows that collagen XVIII is, next to perlecan and agrin, the third basal lamina heparan sulfate proteoglycan (HSPG) and the first collagen/proteoglycan with heparan sulfate side chains. By using monoclonal antibodies to an unidentified HSPG in chick, 14 cDNA clones were isolated from a chick yolk sac library. All clones had a common nucleotide sequence that was homologous to the mRNA sequences of mouse and human collagen XVIII. The deduced amino acid sequence of the chick fragment shows an 83% overall homology with the human and mouse collagen XVIII. Similar to the human and mouse homologue, the chick collagen XVIII mRNA has a size of 4.5 kilobase pairs. In Western blots, collagen XVIII appeared as a smear with a molecular mass of 300 kDa. After treatment with heparitinase, the protein was reduced in molecular mass by 120 kDa to a protein core of 180 kDa. Collagen XVIII has typical features of a collagen, such as its existence, under non-denaturing conditions, as a non-covalently linked oligomer, and a sensitivity of the core protein to collagenase digestion. It also has characteristics of an HSPG, such as long heparitinase-sensitive carbohydrate chains and a highly negative net charge. Collagen XVIII is abundant in basal laminae of the retina, epidermis, pia, cardiac and striated muscle, kidney, blood vessels, and lung. In situ hybridization showed that the main expression of collagen XVIII HSPG in the chick embryo is in the kidney and the peripheral nervous system. As a substrate, collagen XVIII moderately promoted the adhesion of Schwann cells but had no such activity on peripheral nervous system neurons and axons. PMID- 9738010 TI - Insulin stimulation of the fatty acid synthase promoter is mediated by the phosphatidylinositol 3-kinase pathway. Involvement of protein kinase B/Akt. AB - Fatty acid synthase (FAS) is a critical enzyme in de novo lipogenesis. It catalyzes the seven steps in the conversion of malonyl-CoA and acetyl-CoA to palmitate. We have shown that the rate of FAS transcription is induced dramatically when fasted animals are refed with a high carbohydrate, fat-free diet or when streptozotocin-diabetic mice are given insulin. The FAS promoter was up-regulated by insulin through the proximal insulin response sequence containing an E-box motif at the -65-base pair position. Binding of upstream stimulatory factors to the -65 E-box is functionally required for insulin regulation of the FAS promoter. In the present study, we characterized signaling pathways in the insulin stimulation of FAS transcription using specific inhibitors for various signaling molecules and transfecting engineered phosphatidylinositol (PI) 3 kinase subunits and protein kinase B (PKB)/Akt. PD98059 and rapamycin, which inhibit MAP kinase and P70 S6 kinase, respectively, had little effect on the insulin-stimulated FAS promoter activity in 3T3-L1 adipocytes. On the other hand, wortmannin and LY294002, which specifically inactivate PI 3-kinase, strongly inhibited the insulin-stimulated FAS promoter activity. As shown in RNase protection assays, LY294002 also inhibited insulin stimulation of the endogenous FAS mRNA levels in 3T3-L1 adipocytes. Cotransfection of expression vectors for the constitutively active P110 subunit of PI 3-kinase resulted in an elevated FAS promoter activity in the absence of insulin and a loss of further insulin stimulation. Transfecting a dominant negative P85 subunit of PI 3-kinase decreased FAS promoter activity and blocked insulin stimulation. Furthermore, cotransfected wild-type PKB/Akt increased FAS promoter activity in the absence of insulin and a loss of insulin responsiveness of the FAS promoter. On the other hand, kinase-dead PKB/Akt acted in a dominant negative manner to decrease the FAS promoter activity and abolished its insulin responsiveness. These results demonstrate that insulin stimulation of fatty acid synthase promoter is mediated by the PI 3-kinase pathway and that PKB/Akt is involved as a downstream effector. PMID- 9738011 TI - Ikappa Balpha functions through direct contacts with the nuclear localization signals and the DNA binding sequences of NF-kappaB. AB - We have determined the binding energies of complexes formed between Ikappa Balpha and the wild type and mutational variants of three different Rel/NF-kappaB dimers, namely, the p50/p65 heterodimer and homodimers of p50 and p65. We show that although a common mode of interaction exists between the Rel/NF-kappaB dimers and Ikappa Balpha, IkappaB alpha binds the NF-kappaB p50/p65 heterodimer with 60- and 27-fold higher affinity than the p50 and p65 homodimers, respectively. Each of the three flexibly linked segments of the rel homology region of Rel/NF-kappaB proteins (the nuclear localization sequence, the dimerization domain, and the amino-terminal DNA binding domain) is directly engaged in forming the protein/protein interface with the ankyrin repeats and the carboxyl-terminal acidic tail/PEST sequence of Ikappa Balpha. In the cell, Ikappa Balpha functions to retain NF-kappaB in the cytoplasm and inhibit its DNA binding activity. These properties are a result of the direct involvement of the nuclear localization sequences and of the DNA binding region of NF-kappaB in complex with Ikappa Balpha. A model of the interactions in the complex is proposed based on our observations and the crystal structures of Rel/NF-kappaB dimers and the ankyrin domains of related proteins. PMID- 9738012 TI - A functional role for mitochondrial protein kinase Calpha in Bcl2 phosphorylation and suppression of apoptosis. AB - Phosphorylation of Bcl2 at serine 70 may result from activation of a classic protein kinase C (PKC) isoform and is required for functional suppression of apoptosis by Bcl2 in murine growth factor-dependent cell lines (Ito, T., Deng, X., Carr, B., and May, W. S. (1997) J. Biol. Chem. 272, 11671-11673). Human pre-B REH cells express high levels of Bcl2 yet remain sensitive to the chemotherapeutic agents etoposide, cytosine arabinoside, and Adriamycin. In contrast, myeloid leukemia-derived HL60 cells express less than half the level of Bcl-2 but are >10-fold more resistant to apoptosis induced by these drugs. The mechanism responsible for this apparent dichotomy appears to involve a deficiency of mitochondrial PKCalpha since 1) HL60 but not REH cells contain highly phosphorylated Bcl2; 2) PKCalpha is the only classical isoform co-localized with Bcl2 in HL60 but not REH mitochondrial membranes; 3) the natural product and potent PKC activator bryostatin-1 induces mitochondrial localization of PKCalpha in association with Bcl2 phosphorylation and increased REH cell resistance to drug-induced apoptosis; 4) PKCalpha can directly phosphorylate wild-type but not phosphorylation-negative and loss of function S70A Bcl2 in vitro; 5) stable, forced expression of exogenous PKCalpha induces mitochondrial localization of PKCalpha, increased Bcl2 phosphorylation and a >10-fold increase in resistance to drug-induced cell death; and () PKCalpha-transduced cells remain highly sensitive to staurosporine, a potent PKC inhibitor. Furthermore, treatment of the PKCalpha transformants with bryostatin-1 leads to even higher levels of mitochondrial PKCalpha, Bcl2 phosphorylation, and REH cell survival following chemotherapy. While these findings strongly support a role for PKCalpha as a functional Bcl2 kinase that can enhance cell resistance to antileukemic chemotherapy, they do not exclude the possibility that another Bcl2 kinase(s) may also exist. Collectively, these findings identify a functional role for PKCalpha in Bcl2 phosphorylation and in resistance to chemotherapy and suggest a novel target for antileukemic strategies. PMID- 9738013 TI - Transcriptional regulation of the human quinone reductase gene by antiestrogen liganded estrogen receptor-alpha and estrogen receptor-beta. AB - We have previously reported that antiestrogens stimulate quinone reductase (NAD(P)H:(quinone-acceptor) oxidoreductase (QR or NQO1); EC 1.6.99.2) enzymatic activity, an action that may provide protective effects against the toxicity and mutagenicity caused by quinones. We have now investigated the transcriptional regulation of the QR gene by antiestrogens. In transfection experiments employing the 5'-flanking (863-base pair) region of the human QR gene promoter with its electrophile/antioxidant response element (EpRE/ARE) or deleted or mutated constructs, we observe that antiestrogens induced an increase in QR gene promoter reporter activity in estrogen receptor (ER) negative breast cancer and endometrial cancer cells transfected with ER, and this induction by antiestrogens was repressed by estradiol. The stimulation of QR transcriptional activity required the 31-base pair electrophile-responsive region from the human QR gene promoter and a functional ER. Intriguingly, antiestrogens were stronger activators of the QR EpRE via the ER subtype ERbeta than ERalpha. Oligonucleotide gel mobility and antibody shift assays reveal that the ER binds to the EpRE but is only a minor component of the proteins bound to the EpRE in ER-containing MCF 7 breast cancer cells. While binding of ERbeta to the estrogen response element was weaker when compared with ERalpha, ERbeta and ERalpha showed similar binding to the EpRE. Together these findings provide evidence that QR gene regulation by the antiestrogen-occupied ER is mediated by the EpRE-containing region of the human QR gene and indicate that the ER is one of the complex of proteins that binds to the EpRE. In addition, that ERbeta is a more potent activator at EpRE elements than is ERalpha suggests that the different levels of these two receptors in various estrogen target cells could impact importantly on the antioxidant potency of antiestrogens in different target cells. These findings have broad implications regarding the potential beneficial effects of antiestrogens since EpREs mediate the transcriptional induction of numerous genes, including QR, which encode chemoprotective detoxification enzymes. PMID- 9738015 TI - Cloning and deletion mutagenesis of the alpha2 delta calcium channel subunit from porcine cerebral cortex. Expression of a soluble form of the protein that retains [3H]gabapentin binding activity. AB - The anti-epileptic, anti-hyperalgesic, and anxiolytic agent gabapentin (1 (aminomethyl)-cyclohexane acetic acid or Neurontin) has previously been shown to bind with high affinity to the alpha2delta subunit of voltage-dependent calcium channels (Gee, N. S. , Brown, J. P., Dissanayake, V. U. K., Offord, J., Thurlow, R., and Woodruff, G.N. (1996) J. Biol. Chem. 271, 5768-5776). We report here the cloning, sequencing, and deletion mutagenesis of the alpha2delta subunit from porcine brain. The deduced protein sequence has a 95.9 and 98.2% identity to the rat and human neuronal alpha2 delta sequences, respectively. [3H]Gabapentin binds with a KD of 37.5 +/- 10.4 nM to membranes prepared from COS-7 cells transfected with wild-type porcine alpha2 delta cDNA. Six deletion mutants (B-G) that lack the delta polypeptide, together with varying amounts of the alpha2 component, failed to bind [3H]gabapentin. C-terminal deletion mutagenesis of the delta polypeptide identified a segment (residues 960-994) required for correct assembly of the [3H]gabapentin binding pocket. Mutant L, which lacks the putative membrane anchor in the delta sequence, was found in both membrane-associated and soluble secreted forms. The soluble form was not proteolytically cleaved into separate alpha2 and delta chains but still retained a high affinity (KD = 30.7 +/- 8.1 nM) for [3H]gabapentin. The production of a soluble alpha2delta mutant supports the single transmembrane model of the alpha2 delta subunit and is an important step toward the large-scale recombinant expression of the protein. PMID- 9738014 TI - Expression of a dominant interfering dynamin mutant in 3T3L1 adipocytes inhibits GLUT4 endocytosis without affecting insulin signaling. AB - To examine the role of clathrin-coated vesicle endocytosis in insulin receptor signaling and GLUT4 trafficking, we used recombinant adenovirus to express a dominant interfering mutant of dynamin (K44A/dynamin) in 3T3L1 adipocytes. Functional expression of K44A/dynamin, as measured by inhibition of transferrin receptor internalization, did not affect insulin-stimulated insulin receptor autophosphorylation, Shc tyrosine phosphorylation, or mitogen-activated protein kinase activation. Although the tyrosine phosphorylation of insulin receptor substrate-1 was slightly reduced, correlating with a 25% decrease in insulin receptor substrate-1-associated phosphatidylinositol 3-kinase activity, insulin stimulated Akt kinase activation was unaffected. In contrast, expression of K44A/dynamin resulted in the cell-surface accumulation of GLUT4 under basal conditions and an inhibition of GLUT4 endocytosis without affecting insulin stimulated GLUT4 exocytosis. These data demonstrate that disruption of clathrin mediated endocytosis does not significantly perturb insulin receptor signal transduction pathways. Furthermore, K44A/dynamin expression causes an accumulation of GLUT4 at the cell surface, suggesting that GLUT4 vesicles exist in at least two distinct intracellular compartments, one that undergoes continuous recycling and a second that is responsive to insulin. PMID- 9738016 TI - Heat shock factor 1 mediates hemin-induced hsp70 gene transcription in K562 erythroleukemia cells. AB - Transcriptional induction of the hsp70 gene is mediated by heat shock factor 1 (HSF1) rapidly activated upon heat and other stresses. HSF2 has been thought to be responsible for accumulation of HSP70 during hemin-induced differentiation of human K562 erythroleukemia cells because of accompanying acquisition of HSF2 DNA binding activity. However, there has not been any direct evidence for such a functional role of HSF2. The purpose of this study is to clarify the roles of HSF1 and HSF2 in HSP70 induction in hemin-treated K562 cells. We show here that a chimeric polypeptide of HSF2 and GAL4 DNA binding domain (GAL4-BD-HSF2) was unable to induce a GAL4 binding site-containing luciferase reporter gene in response to hemin and that exogenously overproduced HSF2 also failed to increase expression of a heat shock element-containing reporter. On the contrary, expression of a GAL4-BD-HSF1 chimeric protein responded to hemin treatment as well as to heat shock, and transiently overexpressed HSF1 caused hemin-responsive induction of the reporter gene in a dose-dependent manner. These results indicate that HSF1, rather than HSF2, primarily mediates the hemin-induced transcription of the hsp70 gene. PMID- 9738017 TI - Cloning and characterization of the human PAX2 promoter. AB - PAX2, a member of the PAX gene family of developmental transcription factors, is expressed at high levels in the developing eyes, ears, central nervous and urogenital systems, as well as in Wilms' tumor and renal cell carcinoma. Expression of PAX2 in the urogenital system is associated with proliferating cells of the ureteric bud and the differentiating nephrogenic mesenchyme. To date, little is known about the molecular mechanisms controlling the regulation of PAX2 expression. This report describes the cloning and characterization of the human PAX2 gene promoter and localization of the transcription start sites in fetal kidney and Wilms' tumor. We identified two transcription start sites in a Wilms' tumor sample, which were found to be different from that in fetal kidney. The activity of a deletion series of the PAX2 promoter was assessed in NIH-3T3, COS-7, 293, and Madin-Darby canine kidney cells. Although some differences were observed in the activity of each promoter construct, the profile of activity for the promoter fragment series was similar in each experiment, regardless of cell type. The WT1 tumor suppressor protein, which has previously been shown to repress murine Pax2 expression in vitro, was shown to also repress expression from the human PAX2 promoter. PMID- 9738018 TI - Targeting of constitutively active phosphoinositide 3-kinase to GLUT4-containing vesicles in 3T3-L1 adipocytes. AB - Constitutive activation of phosphoinositide 3-kinase (PI3K) stimulates glucose transport and GLUT4 glucose transporter translocation to the plasma membrane in adipocytes. To determine whether a direct interaction of PI3K with GLUT4 containing vesicles (hereafter called GLUT4 vesicles) is important for the effect of insulin on GLUT4 translocation, we targeted constitutively active PI3K to GLUT4 vesicles. We fused the inter-Src homology region 2 of the regulatory p85alpha subunit of PI3K (iSH2) either to a C-terminal sequence of GLUT4 (G4c, amino acids 406-509) or to this region and the N-terminal tail of GLUT4 (G4n, amino acids 1-19), resulting in the fusion proteins iSH2-G4c and G4n-iSH2-G4c, respectively. Coexpression of the fusion proteins or untargeted iSH2 with the catalytic p110alpha subunit of PI3K (p110) in 3T3-L1 adipocytes by adenovirus mediated gene transfer increased total PI3K activity in homogenates 5.0-6.7-fold over nontransduced cells or cells transduced with adenovirus encoding beta galactosidase. In contrast, PI3K activity in GLUT4 vesicles increased 11-13-fold with expression of either targeted construct and p110 but only 2-fold with the untargeted iSH2 and p110, indicating successful targeting of PI3K to GLUT4 vesicles. Both targeted and nontargeted constructs stimulated DNA synthesis to levels greater than insulin, demonstrating that both types of constructs had biologic activity in intact cells. Despite this, untargeted iSH2/p110 coexpression was more effective in stimulating 2-deoxyglucose uptake (6-fold) than either iSH2-G4c/p110 or G4n-iSH2-G4c/p110 coexpression (both 2-fold). Only iSH2/p110 coexpression led to a significant GLUT4 translocation to the plasma membrane. Insulin-stimulated glucose transport was unaffected by any construct. Thus, a direct interaction between PI3K and GLUT4 vesicles is either not required or not sufficient for GLUT4 translocation and stimulation of glucose transport. PMID- 9738019 TI - Serotonin-mediated production of interstitial collagenase by uterine smooth muscle cells requires interleukin-1alpha, but not interleukin-1beta. AB - The activation of the gene for interstitial collagenase in myometrial smooth muscle cells is absolutely dependent upon the presence of serotonin. Our previous studies investigating the mechanisms of this induction demonstrated that the mRNAs of both interleukin-1 (IL-1) isoforms, IL-1alpha and IL-1beta, are induced by serotonin and that the induction of IL-1 is required for the subsequent induction of collagenase. These data provided compelling evidence that serotonin induced IL-1 acts via an autocrine loop in activating the collagenase gene. The experiments described here were designed to examine the potential role of each IL 1 isoform in collagenase production by using neutralizing antisera specific to each isoform of the cytokine. The antisera were examined for their ability to inhibit the serotonin-dependent production of the mRNA for collagenase and of the cytokines themselves. Neutralizing antiserum against IL-1alpha, but not against IL-1beta, inhibited the induction of the mRNA for collagenase and of the mRNAs for both IL-1alpha and IL-1beta. Western analysis indicated that detectable levels of IL-1alpha protein, but not that of IL-1beta, are produced at the time of serotonin-dependent collagenase induction. In contrast, significant levels of IL-1beta protein are detected only when bacterial lipopolysaccharide is added to the cells. Taken together, the results of our study indicate that IL-1alpha, but not IL-1beta, plays an obligatory role in multiple serotonin-mediated gene regulations in the myometrial smooth muscle cell. In addition, the data suggest that IL-1beta production has the potential for modifying myometrial function in pathological settings, particularly that of uterine infection. PMID- 9738020 TI - Role of immunoglobulin-like domains 2-4 of the platelet-derived growth factor alpha-receptor in ligand-receptor complex assembly. AB - Platelet-derived growth factor (PDGF) is a dimeric protein that exerts its effects through tyrosine kinase alpha- and beta-receptors. The extracellular part of each receptor is composed of five Ig-like domains. Recombinant forms of alpha receptor domains 1-4 (alphaRD1-4), 1-3 (alphaRD1-3), and 1 and 2 (alphaRD1-2) were prepared after expression in Chinese hamster ovary cells and were used to study the assembly of soluble ligand-receptor complexes. When incubated with micromolar concentrations of PDGF, both alphaRD1-3 and alphaRD1-4 formed complexes of 1:2 molar composition, i.e. one dimeric PDGF molecule bound two soluble receptors. alphaRD1-3, in contrast to alphaRD1-4, formed detectable 1:1 complexes under conditions of ligand excess. alphaRD1-4 displayed an increased ability to form 1:2 complexes as compared with alphaRD1-3 under conditions of limiting concentrations of ligand. We thus conclude that Ig-like domain 4 mediated receptor-receptor interactions contribute to 1:2 PDGF.alphaRD1-4 complex formation. Since alphaRD1-4 and alphaRD1-3 were equipotent in blocking binding of subnanomolar concentrations of PDGF to cell-surface receptors, we also conclude that this effect is predominantly achieved through formation of Ig-like domain 4 independent 1:1 ligand-receptor complexes. Finally, since alphaRD1-2 bound PDGF BB with high affinity, whereas PDGF-AA was bound only with low affinity, we conclude that Ig-like domain 3 of the PDGF alpha-receptor contains epitopes of particular importance for PDGF-AA binding and that most of the PDGF-BB-binding epitopes reside in Ig-like domains 1 and 2. PMID- 9738021 TI - Molecular cloning and functional expression of a skeletal muscle dihydropyridine receptor from Rana catesbeiana. AB - In skeletal muscle the dihydropyridine receptor is the voltage sensor for excitation-contraction coupling and an L-type Ca2+ channel. We cloned a dihydropyridine receptor (named Fgalpha1S) from frog skeletal muscle, where excitation-contraction coupling has been studied most extensively. Fgalpha1S contains 5600 base pairs coding for 1688 amino acids. It is highly homologous with, and of the same length as, the C-truncated form predominant in rabbit muscle. The primary sequence has every feature needed to be an L-type Ca2+ channel and a skeletal-type voltage sensor. Currents expressed in tsA201 cells had rapid activation (5-10 ms half-time) and Ca2+-dependent inactivation. Although functional expression of the full Fgalpha1S was difficult, the chimera consisting of Fgalpha1S domain I in the rabbit cardiac Ca channel had high expression and a rapidly activating current. The slow native activation is therefore not determined solely by the alpha1 subunit sequence. Its Ca2+ dependent inactivation strengthens the notion that in rabbit skeletal muscle this capability is inhibited by a C-terminal stretch (Adams, B., and Tanabe, T. (1997) J. Gen. Physiol. 110, 379-389). This molecule constitutes a new tool for studies of excitation-contraction coupling, gating, modulation, and gene expression. PMID- 9738022 TI - HBx protein of hepatitis B virus activates Jak1-STAT signaling. AB - The X-gene product (HBx) of the hepatitis B virus plays essential roles in viral replication and the generation of hepatocellular carcinoma. Although the mechanism for HBx action is unclear, HBx may exert its pleiotropic functions through the stimulation of signal transduction pathways including the Ras/mitogen activated protein kinase cascade and/or inactivation of the p53 function. Here, we investigated whether HBx has the ability to activate the Jak-STAT signaling pathway. As a first step, we established stable cell lines constitutively expressing HBx. In these HBx-expressing stable cells, the tyrosine phosphorylation of various STATs, including STAT3 and -5, was constitutively enhanced by HBx, and the concomitant increase in STAT-dependent DNA binding and transcriptional activation was observed. Furthermore, HBx specifically elevated tyrosine phosphorylation and in vitro kinase activity of Jak1, but not Jak2 or Tyk2, through protein to protein interaction with Jak1. These results clearly establish HBx as the inducer of the Jak-STAT signaling pathway, and at the same time, HBx-mediated Jak-STAT activation may provide a novel mechanism for the pleiotropic functions of HBx, including transformation and promiscuous transcriptional activation. PMID- 9738023 TI - Characterization of the Escherichia coli RNA 3'-terminal phosphate cyclase and its sigma54-regulated operon. AB - The RNA 3'-terminal phosphate cyclase catalyzes the ATP-dependent conversion of the 3'-phosphate to the 2',3'-cyclic phosphodiester at the end of various RNA substrates. Recent cloning of a cDNA encoding the human cyclase indicated that genes encoding cyclase-like proteins are conserved among Eucarya, Bacteria, and Archaea. The protein encoded by the Escherichia coli gene was overexpressed and shown to have the RNA 3'-phosphate cyclase activity (Genschik, P., Billy, E., Swianiewicz, M., and Filipowicz, W. (1997) EMBO J. 16, 2955-2967). Analysis of the requirements and substrate specificity of the E. coli protein, presented in this work, demonstrates that properties of the bacterial and human enzymes are similar. ATP is the best cofactor (Km = 20 microM), whereas GTP (Km = 100 microM) and other nucleoside triphosphates (NTPs) act less efficiently. The enzyme undergoes nucleotidylation in the presence of [alpha-32P]ATP and, to a lesser extent, also in the presence of other NTPs. Comparison of 3'-phosphorylated oligoribonucleotides and oligodeoxyribonucleotides of identical sequence demonstrated that the latter are at least 300-fold poorer substrates for the enzyme. The E. coli cyclase gene, named rtcA, forms part of an uncharacterized operon containing two additional open reading frames (ORFs). The ORF positioned immediately upstream, named rtcB, encodes a protein that is also highly conserved between Eucarya, Bacteria, and Archaea. Another ORF, called rtcR, is positioned upstream of the rtcA/rtcB unit and is transcribed in the opposite direction. It encodes a protein having features of sigma54-dependent regulators. By overexpressing the N-terminally truncated form of RtcR, we demonstrate that this regulator indeed controls expression of rtcA and rtcB in a sigma54-dependent manner. Also consistent with the involvement of sigma54, the region upstream of the transcription start site of the rtcA/rtcB mRNA contains the -12 and -24 elements, TTGCA and TGGCA, respectively, characteristic of sigma54-dependent promoters. The cyclase gene is nonessential as demonstrated by knockout experiments. Possible functions of the cyclase in RNA metabolism are discussed. PMID- 9738024 TI - Evaluation of commercially available Helicobacter pylori serology kits: a review. PMID- 9738025 TI - Improved amplification of microbial DNA from blood cultures by removal of the PCR inhibitor sodium polyanetholesulfonate. AB - Molecular methods are increasingly used to identify microbes in clinical samples. A common technical problem with PCR is failed amplification due to the presence of PCR inhibitors. Initial attempts at amplification of the bacterial 16S rRNA gene from inoculated blood culture media failed for this reason. The inhibitor persisted, despite numerous attempts to purify the DNA, and was identified as sodium polyanetholesulfonate (SPS), a common additive to blood culture media. Like DNA, SPS is a high-molecular-weight polyanion that is soluble in water but insoluble in alcohol. Accordingly, SPS tends to copurify with DNA. An extraction method was designed for purification of DNA from blood culture media and removal of SPS. Blood culture media containing human blood and spiked with Escherichia coli was subjected to an organic extraction procedure with benzyl alcohol, and removal of SPS was documented spectrophotometrically. Successful amplification of the extracted E. coli 16S rRNA gene was achieved by adding 5 microliter of undiluted processed sample DNA to a 50-microliter PCR mixture. When using other purification methods, the inhibitory effect of SPS could be overcome only by dilution of these samples. By our extraction technique, even uninoculated blood culture media were found to contain bacterial DNA when they were subjected to broad-range 16S rRNA gene consensus PCR. We conclude that the blood culture additive SPS is a potent inhibitor of PCR, is resistant to removal by traditional DNA purification methods, but can be removed by a benzyl alcohol extraction protocol that results in improved PCR performance. PMID- 9738026 TI - Detection of resistance to amphotericin B among Cryptococcus neoformans clinical isolates: performances of three different media assessed by using E-test and National Committee for Clinical Laboratory Standards M27-A methodologies. AB - Although reliable detection of resistance in vitro is critical to the overall performance of any susceptibility testing method, the recently released National Committee for Clinical Laboratory Standards M27-A methodology for susceptibility testing of yeasts discriminates poorly between resistant and susceptible isolates of Candida spp. We have previously shown that both substitution of antibiotic medium 3 for RPMI 1640 medium in the microdilution variant of the M27-A method and use of the E-test agar diffusion methodology permit detection of amphotericin B-resistant Candida isolates. To determine the relevance of these observations to Cryptococcus neoformans, we have evaluated the performances of both the M27-A and the E-test methodologies with this yeast using three different media (RPMI 1640 medium, antibiotic medium 3, and yeast nitrogen base). As with Candida, we found that only antibiotic medium 3 permitted consistent detection of resistant isolates when testing was performed in broth by the M27-A method. When testing was performed by the E-test agar diffusion method, both RPMI 1640 medium and antibiotic medium 3 agar permitted ready detection of the resistant isolates. Reading of the results after 48 h of incubation was required for testing in broth by the M27-A method, while the MIC could be determined after either 48 or 72 h when the agar diffusion method was used. PMID- 9738028 TI - Molecular epidemiology of recent belgian isolates of Neisseria meningitidis serogroup B. AB - In Belgium an increase in the incidence of meningococcal disease has been noted since the early 1990s. Four hundred twenty clinical strains isolated during the period from 1990 to 1995, along with a set of 30 European reference strains, and 20 Dutch isolates were examined by random-primer and repetitive-motif-based PCR. A subset was investigated by multilocus enzyme electrophoresis and pulsed-field gel electrophoresis. The data were compared with results obtained by serotyping (M. Van Looveren, F. Carion, P. Vandamme, and H. Goossens, Clin. Microbiol. Infect. 4:224-228, 1998). Both phenotypic and molecular epidemiological data suggest that the lineage III of Neisseria meningitidis, first encountered in The Netherlands in about 1980, has been introduced in Belgium. The epidemic clone, as defined by oligonucleotide D8635-primed PCR, encompasses mainly phenotypes B:4:P1.4 and B:nontypeable:P1.4, but strains with several other phenotypes were also encountered. Therefore, serotyping alone would underestimate the prevalence of the epidemic clone. PMID- 9738027 TI - Human exposure to a granulocytic Ehrlichia and other tick-borne agents in Connecticut. AB - Indirect fluorescent-antibody (IFA) staining methods with Ehrlichia equi (MRK or BDS strains) and Western blot analyses containing a human granulocytic ehrlichiosis (HGE) agent (NCH-1 strain) were used to confirm probable human cases of infection in Connecticut during 1995 and 1996. Also included were other tests for Ehrlichia chaffeensis, the agent of human monocytic ehrlichiosis (HME), Babesia microti, and Borrelia burgdorferi. Thirty-three (8.8%) of 375 patients who had fever accompanied by marked leukopenia or thrombocytopenia were serologically confirmed as having HGE. Western blot analyses of a subset of positive sera confirmed the results of the IFA staining methods for 15 (78.9%) of 19 seropositive specimens obtained from different persons. There was frequent detection of antibodies to a 44-kDa protein of the HGE agent. Serologic testing also revealed possible cases of Lyme borreliosis (n = 142), babesiosis (n = 41), and HME (n = 21). Forty-seven (26.1%) of 180 patients had antibodies to two or more tick-borne agents. Therefore, when one of these diseases is clinically suspected or diagnosed, clinicians should consider the possibility of other current or past tick-borne infections. PMID- 9738029 TI - Restriction fragment length polymorphism analysis of some flagellin genes of Salmonella enterica. AB - Salmonellae often have the ability to express two different flagellar antigen specificities (phase 1 and phase 2). At the cell level, only one flagellar phase is expressed at a time. Two genes, fliC, encoding phase-1 flagellin, and fljB, encoding phase-2 flagellin, are alternatively expressed. Flagellin genes from 264 serovars of Salmonella enterica were amplified by two phase-specific PCR systems. Amplification products were subjected to restriction fragment length polymorphism (RFLP) analysis by using endonucleases HhaI and HphI. RFLP with HhaI and HphI yielded 64 and 42 different restriction profiles, respectively, among 329 flagellin genes coding for 26 antigens. The phase-1 gene showed 46 patterns with HhaI and 30 patterns with HphI. The phase-2 gene showed 23 patterns with HhaI and 17 patterns with HphI. When the data from both enzymes were combined, 116 patterns were obtained: 74 for fliC, 47 for fljB, and 5 shared by both genes. Of these combined patterns, 80% were specifically associated with one flagellar antigen and 20% were associated with more than one antigen. Each flagellar antigen was divided into 2 to 18 different combined patterns. In the sample of strains used, determination of the phase-1 and phase-2 flagellin gene RFLP, added to the knowledge of the O antigen, allowed identification of all diphasic serovars. Overall, the diversity uncovered by flagellin gene RFLP did not precisely match that evidenced by flagellar agglutination. PMID- 9738030 TI - PCR for detection and identification of Abiotrophia spp. AB - Members of the genus Abiotrophia, formerly known as nutritionally variant streptococci, are important pathogens causing septicemia and endocarditis. Cultivation and biochemical differentiation of Abiotrophia spp. are often difficult. Based on 16S rRNA sequences, two PCR assays for detection and identification of Abiotrophia spp. were developed. The first PCR assay was positive for all Abiotrophia spp. Subsequently performed restriction fragment length polymorphism analysis allowed the verification of the PCR amplicons and the differentiation of the three species. The second PCR assay was positive only for A. elegans, the most fastidious species of Abiotrophia. Both PCR assays were shown to be specific and sensitive and should facilitate the identification of Abiotrophia spp. PMID- 9738031 TI - Massilia timonae gen. nov., sp. nov., isolated from blood of an immunocompromised patient with cerebellar lesions. AB - A fastidious, slowly growing, strictly aerobic, gram-negative bacterium was isolated from a culture of blood from a 25-year-old man with common variable immunodeficiency. The man had been admitted to hospital with febrile progressive cerebellar ataxia. The use of standard phenotypic schemes did not lead to identification, but sequence analysis demonstrated that the 16S rRNA gene of the isolate was most similar to those of the environmental bacteria Duganella zoogloeoides (formerly Zoogloea ramigera 115) and Telluria mixta. Further characterization of the bacterium by biochemical analysis, electron microscopy, G+C content estimation, and fatty acid analysis demonstrated significant differences between the bacterium and D. zoogloeoides and Telluria species; thus, we propose it as a new taxon with the name Massilia timonae gen. nov., sp. nov. PMID- 9738032 TI - Clinical evaluation of the automated COBAS AMPLICOR MTB assay for testing respiratory and nonrespiratory specimens. AB - We evaluated the COBAS AMPLICOR PCR system (Roche Diagnostics) for the routine detection of Mycobacterium tuberculosis complex (MTBC) in clinical specimens. Diagnostic culture, considered as the reference method, was performed with BACTEC, Lowenstein-Jensen, Stonebrink, and Kirchner media. Occasionally MB-Redox, ESP, or MGIT medium was also used. A total of 643 respiratory and 506 nonrespiratory specimens collected from 807 patients were investigated. Of the 95 culture-positive specimens, 80 were COBAS AMPLICOR MTB positive, and of the 1,054 culture-negative specimens, 1,044 were COBAS AMPLICOR MTB negative. After resolving discrepancies by review of the medical history, the overall sensitivity, specificity, and positive and negative predictive values for the COBAS AMPLICOR MTB assay, respectively, were 83.5, 98.8, 86.7, and 98.6% compared to those of diagnostic culture. In smear-positive specimens, the sensitivity of the COBAS AMPLICOR MTB assay was 96%, versus 48% for smear-negative specimens. No significant differences in the test performance between respiratory and nonrespiratory specimens were observed. The overall inhibition rate was less than 2%, excluding stool specimens. The clear advantages of the COBAS AMPLICOR PCR system are standardized procedures and reagents for specimen processing as well as an internal control for reliable monitoring of PCR inhibitors. By simplifying the work flow through a completely automated amplification and amplicon detection procedure, the COBAS AMPLICOR PCR system proved itself as a very useful component for routine diagnostic procedures. PMID- 9738033 TI - Prospective investigation of cryptic outbreaks of Salmonella agona salmonellosis. AB - The number of Salmonella agona isolates reported annually in Texas from 1992 through 1994 ranged from 14 to 21. An increase in incidence of S. agona infections was noted in the fall of 1995. Pulsed-field gel electrophoresis (PFGE) analysis identified prospectively two possible cryptic outbreaks caused by an indistinguishable strain which was isolated from 18 of 59 patients who were culture positive from March through December 1995. These 18 patients had onset of illness from 20 May through 3 October 1995. Eight individuals resided in the Austin area, eight resided in San Antonio, and two resided in Houston; none had attended a common social gathering or owned common pets. Six patients in San Antonio and one patient from Houston recalled eating food items from the same Mexican food restaurant in San Antonio. S. agona organisms with the same PFGE profile were isolated from machacado, an air-dried, raw beef product prepared at the restaurant. The machacado had been shredded in a kitchen blender which was the probable source for cross-contamination of other food items. Five patients in Austin reported eating at a popular Mexican food restaurant in Austin. Improperly prepared machacado also may have been served at the Austin restaurant; however, sufficient quantities of machacado were not available for analysis. PFGE was essential in determining whether the cases constituted outbreaks and was invaluable in guiding the epidemiological investigation. PMID- 9738034 TI - Rapid shell vial culture technique for detection of enteroviruses and adenoviruses in fecal specimens: comparison with conventional virus isolation method. AB - Detection of enteroviruses and adenoviruses mainly in fecal specimens by rapid culture with inoculation onto cell monolayers in flat-bottom tubes by centrifugation and immunofluorescence staining with genus-specific monoclonal antibodies was compared with that by the conventional virus isolation procedure. For both conventional culture and shell vial culture human lung fibroblast cells and tertiary monkey kidney cells were used. For enterovirus detection, 979 clinical specimens (916 stool specimens, 56 cerebrospinal fluid specimens, and 7 nasopharyngeal swabs) were used. Conventional culture detected 74 enterovirus isolates. A cytopathic effect compatible with the presence of an enterovirus after 3 days of incubation occurred in 25 of the 74 (34%) specimens that eventually became positive. The detection rate for enteroviruses by rapid cell culture after 2 to 3 days of incubation was 42 of 74 (57%). The genus-specific enterovirus monoclonal antibody did not react with strains of echovirus types 22 and 23 or enterovirus type 71. Rapid cell culture for the detection of adenoviruses was performed with 567 clinical specimens (536 stool specimens, 25 cerebrospinal fluid specimens, and 6 miscellaneous specimens), in which 42 adenoviruses were found by conventional culture. Nine of the 42 (21%) adenovirus isolates were detected by conventional culture within 3 days after inoculation, whereas 21 (50%) were found by rapid cell culture within 2 to 3 days. Only two of the nine specimens found to be positive for the enteric adenovirus type 41 by conventional culture as well by a type-specific enzyme-linked immunosorbent assay (ELISA) tested positive by rapid cell culture. In conclusion, the rapid shell vial assay allows the early detection and identification of enteroviruses and adenoviruses in clinical specimens but is markedly less sensitive than the conventional isolation procedure according to the eventual results of the conventional isolation procedure. Conventional cell culture remains a prerequisite for serotyping of enteroviral isolates. On the basis of the results for adenovirus type 41, the rapid detection of adenoviruses was not considered to be useful for the detection of clinically relevant adenoviruses in fecal samples. PMID- 9738035 TI - Prevalence of Candida dubliniensis isolates in a yeast stock collection. AB - To establish the historical prevalence of the novel yeast species Candida dubliniensis, a survey of 2,589 yeasts originally identified as Candida albicans and maintained in a stock collection dating back to the early 1970s was undertaken. A total of 590 yeasts, including 93 (18.5%) beta-glucosidase-negative isolates among 502 isolates that showed abnormal colony colors on a differential chromogenic agar and 497 other isolates, were subjected to DNA fingerprinting with the moderately repetitive sequence Ca3. On this basis, 53 yeasts were reidentified as C. dubliniensis (including the C. dubliniensis type strain, included as a blind control in the panel of yeasts). The 52 newly found isolates came from 36 different persons, and a further 3 C. dubliniensis isolates were detected by DNA fingerprinting of previously untested isolates from one of these individuals. The prevalence of C. dubliniensis among yeasts in oral and fecal samples was significantly higher than that among yeasts from other anatomical sites and was significantly higher among human immunodeficiency virus (HIV) infected individuals than among known or presumed HIV-negative individuals. However, a single vaginal isolate and two oral isolates from healthy volunteers confirmed that the species is restricted neither to gastrointestinal sites nor to patients with overt disease. The oldest examples of C. dubliniensis were from oral samples of three patients in the United Kingdom in 1973 and 1975. In comparison with age-matched control isolates of C. albicans, the C. dubliniensis isolates showed slightly higher levels of susceptibility in vitro to amphotericin B and flucytosine and slightly lower levels of susceptibility to three azole antifungal agents. PMID- 9738036 TI - Fluconazole susceptibility testing of Cryptococcus neoformans: comparison of two broth microdilution methods and clinical correlates among isolates from Ugandan AIDS patients. AB - We compared the yeast nitrogen base (YNB) broth microdilution method with the National Committee for Clinical Laboratory Standards (NCCLS) M27-A microdilution reference method for measuring the in vitro susceptibility of Cryptococcus neoformans isolates to fluconazole. A total of 149 isolates of C. neoformans var. neoformans from Ugandan AIDS patients was tested by both methods. An overall agreement of 88% between the two microdilution methods was observed. All isolates grew well in both RPMI 1640 and YNB media, and MICs could be read after 48 h of incubation by both methods. The range of fluconazole MICs obtained with the YNB method was broader than that obtained with the NCCLS method. The extended range of MICs provided by the YNB method may be of clinical value, as it appears that the clinical outcome may be better among patients infected with strains inhibited by lower concentrations of fluconazole as determined by the YNB method. The YNB method appears to be a viable option for testing C. neoformans against fluconazole. PMID- 9738037 TI - A nested-PCR-based schizodeme method for identifying Leishmania kinetoplast minicircle classes directly from clinical samples and its application to the study of the epidemiology of Leishmania tropica in Pakistan. AB - A nested PCR was developed to amplify the variable region of the kinetoplast minicircles of all Leishmania species which infect mammals. Each Leishmania parasite contains approximately 10,000 kinetoplast DNA minicircles, which are unequally distributed among approximately 10 minicircle classes. The PCR primers were designed to bind within the 120-bp conserved region which is common to all minicircle classes; the remaining approximately 600 bp of each minicircle is highly conserved within each minicircle class but highly divergent between classes. The nested PCR generated a strong signal from a minimum of 0.1 fg of Leishmania DNA. Restriction digests of the amplicons from the highest dilutions suggested that minicircles from only a limited number of minicircle classes had acted as template in the reaction. One PCR product was directly sequenced and found to be derived from only one minicircle class. Since the primers amplify all minicircle classes, this indicated that as little as 1/10 of one Leishmania parasite was present in the PCR template. This demonstrated that the nested PCR achieved very nearly the maximum theoretically possible sensitivity and is therefore a potentially useful method for diagnosis. The nested PCR was tested for sensitivity on 20 samples from patients from the Timargara refugee camp, Pakistan. Samples were collected by scraping out a small amount of tissue with a scalpel from an incision at the edge of the lesion; the tissue was smeared on one microscope slide and placed in a tube of 4 M guanidine thiocyanate, in which the sample was stable for at least 1 month. DNA for PCR was prepared by being bound to silica in the presence of 6 M guanidine thiocyanate; washed in guanidine thiocyanate, ethanol, and acetone; and eluted with 10 mM Tris-HCl. PCR products of the size expected for Leishmania tropica were obtained from 15 of the 20 samples in at least one of three replicate reactions. The negative samples were from lesions that had been treated with glucantime or were over 6 months old, in which parasites are frequently scanty. This test is now in routine use for the detection and identification of Leishmania parasites in our clinical laboratory. Fingerprints produced by restriction digests of the PCR products were defined as complex or simple. There were no reproducible differences between the complex restriction patterns of the kinetoplast DNA of any of the parasites from Timargara camp with HaeIII and HpaII. The simple fingerprints were very variable and were interpreted as being the product of PCR on a limited subset of minicircle classes, and consequently, it was thought that the variation was determined by the particular minicircle classes that had been represented in the template. The homogeneity of the complex fingerprints suggests that the present epidemic of cutaneous leishmaniasis in Timargara camp may be due to the spread of a single clone of L. tropica. PMID- 9738038 TI - Direct antimicrobial susceptibility testing of gram-negative bacilli in blood cultures by an electrochemical method. AB - Nonfastidious aerobic gram-negative bacilli (GNB) are commonly isolated from blood cultures. The feasibility of using an electrochemical method for direct antimicrobial susceptibility testing of GNB in positive blood cultures was evaluated. An aliquot (10 microliter) of 1:10-diluted positive blood cultures containing GNB was inoculated into the Bactometer module well (bioMerieux Vitek, Hazelwood, Mo.) containing 1 ml of Mueller-Hinton broth supplemented with an antibiotic. Susceptibility tests were performed in a breakpoint broth dilution format, with the results being categorized as resistant, intermediate, or susceptible. Seven antibiotics (ampicillin, cephalothin, gentamicin, amikacin, cefamandole, cefotaxime, and ciprofloxacin) were used in this study, with each agent being tested at the two interpretive breakpoint concentrations. The inoculated modules were incubated at 35 degreesC, and the change in impedance in each well was continuously monitored for 24 h by the Bactometer. The MICs of the seven antibiotics for each blood isolate were also determined by the standardized broth microdilution method. Of 146 positive blood cultures (1,022 microorganism antibiotic combinations) containing GNB tested by the direct method, the rates of very major, major, and minor errors were 0, 1.1, and 2.5%, respectively. The impedance method was simple; no centrifugation, preincubation, or standardization of the inocula was required, and the susceptibility results were normally available within 3 to 6 h after inoculation. The rapid method may allow proper antimicrobial treatment almost 30 to 40 h before the results of the standard methods are available. PMID- 9738039 TI - Direct identification of Vibrio vulnificus in clinical specimens by nested PCR. AB - This study was performed to establish optimal nested PCR conditions and a high yield DNA extraction method for the direct identification of Vibrio vulnificus in clinical specimens. We designed two sets of primers targeting the V. vulnificus hemolysin/cytolysin gene. The target of the first primer set (P1-P2; sense, 5' GAC-TAT-CGC-ATC-AAC-AAC-CG-3', and antisense, 5'-AGG-TAG-CGA-GTA-TTA-CTG-CC-3', respectively) is a 704-bp DNA fragment. The second set (P3-P4; sense, 5'-GCT-ATT TCA-CCG-CCG-CTC-AC-3', and antisense, 5'-CCG-CAG-AGC-CGT-AAA-CCG-AA-3', respectively) amplifies an internal 222-bp DNA fragment. We developed a direct DNA extraction method that involved boiling the specimen pellet in a 1 mM EDTA 0.5% Triton X-100 solution. The new DNA extraction method was more sensitive and reproducible than other conventional methods. The DNA extraction method guaranteed sensitivity as well, even when V. vulnificus cells were mixed with other bacteria such as Escherichia coli or Staphylococcus aureus. The nested PCR method could detect as little as 1 fg of chromosomal DNA and single CFU of V. vulnificus. We applied the nested PCR protocol to a total of 39 serum specimens and bulla aspirates from septicemic patients. Seventeen (94.4%) of the 18 V. vulnificus culture-positive specimens were positive by the nested PCR. Eight (42.1%) of the 19 culture-negative samples gave positive nested PCR results. PMID- 9738040 TI - Prolonged replication of a type 1 vaccine-derived poliovirus in an immunodeficient patient. AB - VP1 sequences were determined for poliovirus type 1 isolates obtained over a 189 day period from a poliomyelitis patient with common variable immunodeficiency syndrome (a defect in antibody formation). The isolate from the first sample, taken 11 days after onset of paralysis, contained two poliovirus populations, differing from the Sabin 1 vaccine strain by approximately 10%, differing from diverse type 1 wild polioviruses by 19 to 24%, and differing from each other by 5.5% of nucleotides. Specimens taken after day 11 appeared to contain only one major poliovirus population. Evolution of VP1 sequences at synonymous third-codon positions occurred at an overall rate of approximately 3.4% per year over the 189 day period. Assuming this rate to be constant throughout the period of infection, the infection was calculated to have started approximately 9.3 years earlier. This estimate is about the time (6. 9 years earlier) the patient received his last oral poliovirus vaccine dose, approximately 2 years before the diagnosis of immunodeficiency. These findings may have important implications for the strategy to eliminate poliovirus immunization after global polio eradication. PMID- 9738041 TI - Incidence of Toxoplasma gondii infection in 35,940 pregnant women in Norway and pregnancy outcome for infected women. AB - From 1992 to 1994 a screening program for detection of specific Toxoplasma gondii antibodies involving 35,940 pregnant women was conducted in Norway. For women with serological evidence of primary T. gondii infection, amniocentesis and antiparasitic treatment were offered. The amniotic fluid was examined for T. gondii by PCR and mouse inoculation to detect fetal infection. Infants of infected mothers had clinical and serological follow-up for at least 1 year to detect congenital infection. Of the women 10.9% were infected before the onset of pregnancy. Forty-seven women (0.17% among previously noninfected women) showed evidence of primary infection during pregnancy. The highest incidence was detected (i) among foreign women (0.60%), (ii) in the capital city of Oslo (0.46%), and (iii) in the first trimester (0.29%). Congenital infection was detected in 11 infants, giving a transmission rate of 23% overall, 13% in the first trimester, 29% in the second, and 50% in the third. During the 1-year follow-up period only one infant, born to an untreated mother, was found to be clinically affected (unilateral chorioretinitis and loss of vision). At the beginning of pregnancy 0.6% of the previously uninfected women were falsely identified as positive by the Platelia Toxo-IgM test, the percentage increasing to 1.3% at the end of pregnancy. Of the women infected prior to pregnancy 6.8% had persisting specific immunoglobulin M (IgM). A positive specific-IgM result had a low predictive value for identifying primary T. gondii infection. PMID- 9738043 TI - Outbreak of Pseudomonas fluorescens bacteremia among oncology patients. AB - From 7 to 24 March 1997, four patients developed Pseudomonas fluorescens bacteremia at the hospital; one on the oncology ward and the other three in the chemotherapy room. These patients all had underlying malignancies and had the Port-A-Cath (Smiths Industries Medical Systems, Deltec, Inc., St. Paul, Minn.) implants. Three patients had primary bacteremia, and one had Port-A-Cath-related infection. None of these patients had received a blood transfusion before the episodes of bacteremia. All patients recovered: two received antimicrobial agents with in vitro activity against the isolates, and the other two did not have any antibiotic treatment. A total of eight blood isolates were recovered from these patients during the febrile episodes that occurred several minutes after the infusion of chemotherapeutic agents via the Port-A-Cath. These isolates were initially identified as P. fluorescens or Pseudomonas putida (four), Burkholderia (Ralstonia) pickettii (three), and a non-glucose-fermenting gram-negative bacillus (one) by routine biochemical methods and the Vitek GNI card. These isolates were later identified as P. fluorescens on the basis of the characteristic cellular fatty acid chromatogram and the results of supplemental biochemical tests. The identification of identical antibiotypes by the E test and the random amplified polymorphic DNA patterns generated by arbitrarily primed PCR of the isolates showed that the outbreak was caused by a single clone of P. fluorescens. Surveillance cultures of the possibly contaminated infusion fluids and disinfectants, which were performed 7 days after recognition of the last infected patient, failed to isolate P. fluorescens. This report of a small outbreak caused by P. fluorescens suggests that timely, accurate identification of unusual nosocomial pathogens is crucial for early initiation of an epidemiological investigation and timely control of an outbreak. PMID- 9738042 TI - Development of specific immunoglobulins G, M, and A following primary Toxoplasma gondii infection in pregnant women. AB - The development of specific antibodies following primary Toxoplasma gondii infection during pregnancy was assessed by six different antibody assays: dye test, Platelia Toxo-IgG, Toxo-Screen DA IgG, Platelia Toxo-IgM, Toxo-ISAGA IgM, and Platelia Toxo-IgA. A total of 126 sera from 27 pregnant women, for whom the time of acquisition of infection could be estimated fairly accurately, were included. All tests showed great individual variation in the peak amounts of antibodies detected. The times elapsed after infection until the peak was reached also varied greatly from individual to individual: the ranges were 2 to 21 weeks for the dye test, 4 to 36 weeks for Platelia Toxo-IgG, 4 to 30 weeks for Toxo Screen DA IgG, 2 to 18 weeks for Platelia Toxo-IgM, 1 to 6 weeks for Toxo-ISAGA IgM, and 2 to 21 weeks for Platelia Toxo-IgA. In the early phase of the infection the dye test and the specific-IgM tests were the most sensitive. Toxo-Screen DA IgG was more sensitive than Platelia Toxo-IgG in the acute phase, while Platelia Toxo-IgA was clearly the least sensitive assay. Of the sera collected 21 to 52 weeks after infection, all were positive by the dye test, all except one (which was negative by Platelia Toxo-IgG) were positive by the specific-IgG tests, approximately 80% were positive by the IgM tests, and 45% were positive by the IgA test. Due to the great individual variation it seems impossible to estimate when the infection occurred based on results obtained from a single serum, and it may even be difficult to assess when a titer increase in paired sera is detectable unless the first sample is only marginally positive. As a diagnostic criterion a dye test titer of >/=300 IU/ml has a low sensitivity for recent primary infection. PMID- 9738044 TI - Molecular genetic variation in Emmonsia crescens and Emmonsia parva, etiologic agents of adiaspiromycosis, and their phylogenetic relationship to Blastomyces dermatitidis (Ajellomyces dermatitidis) and other systemic fungal pathogens. AB - Emmonsia crescens, an agent of adiaspiromycosis, Blastomyces dermatitidis, the agent of blastomycosis, and Histoplasma capsulatum, the agent of histoplasmosis, are known to form meiotic (sexual) stages in the ascomycete genus Ajellomyces (Onygenaceae, Onygenales), but no sexual stage is known for E. parva, the type species of the genus Emmonsia. To evaluate relationships among members of the putative Ajellomyces clade, large-subunit ribosomal and internal transcribed spacer region DNA sequences were determined from PCR-amplified DNA fragments. Sequences were analyzed phylogenetically to evaluate the genetic variation within the genus Emmonsia and evolutionary relationships to other taxa. E. crescens and E. parva are distinct species. E. crescens isolates are placed into two groups that correlate with their continents of origin. Considerable variation occurred among isolates previously classified as E. parva. Most isolates are placed into two closely related groups, but the remaining isolates, including some from human sources, are phylogenetically distinct and represent undescribed species. Strains of B. dermatitidis are a sister species of E. parva. Paracoccidioides brasiliensis and Histoplasma capsulatum are ancestral to most Emmonsia isolates, and P. brasiliensis, which has no known teleomorph, falls within the Ajellomyces clade. PMID- 9738046 TI - Prospective evaluation of criteria for microbiological diagnosis of prosthetic joint infection at revision arthroplasty. The OSIRIS Collaborative Study Group. AB - A prospective study was performed to establish criteria for the microbiological diagnosis of prosthetic joint infection at elective revision arthroplasty. Patients were treated in a multidisciplinary unit dedicated to the management and study of musculoskeletal infection. Standard multiple samples of periprosthetic tissue were obtained at surgery, Gram stained, and cultured by direct and enrichment methods. With reference to histology as the criterion standard, sensitivities, specificities, and likelihood ratios (LRs) were calculated by using different cutoffs for the diagnosis of infection. We performed revisions on 334 patients over a 17-month period, of whom 297 were evaluable. The remaining 37 were excluded because histology results were unavailable or could not be interpreted due to underlying inflammatory joint disease. There were 41 infections, with only 65% of all samples sent from infected patients being culture positive, suggesting low numbers of bacteria in the samples taken. The isolation of an indistinguishable microorganism from three or more independent specimens was highly predictive of infection (sensitivity, 65%; specificity, 99.6%; LR, 168.6), while Gram staining was less useful (sensitivity, 12%; specificity, 98%; LR, 10). A simple mathematical model was developed to predict the performance of the diagnostic test. We recommend that five or six specimens be sent, that the cutoff for a definite diagnosis of infection be three or more operative specimens that yield an indistinguishable organism, and that because of its low level of sensitivity, Gram staining should be abandoned as a diagnostic tool at elective revision arthroplasty. PMID- 9738045 TI - Genetic and serological evidence for multiple instances of unrecognized transmission of hepatitis C virus in hemodialysis units. AB - We investigated the unrecognized patient-to-patient transmission of hepatitis C virus (HCV) in hemodialysis units by performing phylogenetic and serological analyses of hypervariable region 1 (HVR1) of HCV. Of the 62 patients in one center, 11 were positive for HCV RNA. A total of 24 HVR1 sequences, including the minor population of sequences of HCV isolates, from each patient were closely related and classified into five clusters by phylogenetic analysis. Of the 11 patients, 5 were infected with multiple clusters of HCV. Two patients were infected with HCV during an 18-month interval between examinations, and these HVR1 sequences fell into one of the five clusters. In another hemodialysis center, 5 of the 20 patients were HCV RNA positive, and two HVR1 sequences were found to be closely related and phylogenetically derived from the same cluster. The antibody responses of these patients to the HVR1 peptides representative of the genetic clusters revealed exactly the same clustering as that shown by phylogenetic analysis. These findings suggest that phylogenetic and serological analyses of HVR1 sensitively detect unrecognized and multiple transmission of HCV occurring within the same room in hemodialysis centers. Fingerprinting analyses using hypervariable regions of infectious agents are useful in identifying the precise route of transmission of infection. PMID- 9738047 TI - Evaluation of the ESP culture system II for testing susceptibilities of Mycobacterium tuberculosis isolates to four primary antituberculous drugs. AB - The reliability of the ESP Culture System II (herein referred to as ESP II) for testing susceptibilities of Mycobacterium tuberculosis isolates to isoniazid, rifampin, ethambutol, and streptomycin was evaluated by comparing results to those of the method of proportion (MOP), which was considered the reference method, for 20 clinical isolates and 30 challenge strains provided by the Centers for Disease Control and Prevention (CDC). Clinical isolates also were tested with the BACTEC TB 460 system; these results agreed with those obtained by the MOP for all isolates and all drugs, except the high concentration of isoniazid, for which agreement was 95%. After resolution of discrepancies, levels of agreement between ESP II and MOP for the clinical isolates were 95 and 100%, respectively, for the low and high concentrations of isoniazid, 100% for rifampin and ethambutol, and 95% for streptomycin. For the 30 challenge isolates, ESP II results for both concentrations of isoniazid agreed with the expected results in all cases, whereas agreement was 93% for both rifampin and streptomycin and 90% for ethambutol. All discrepancies with the CDC isolates were due to failure of ESP II to correctly classify resistant strains. By testing isolates yielding discrepant ethambutol and streptomycin results with a lower concentration of both drugs in the ESP II system, agreement increased to 93% for ethambutol and 100% for streptomycin. For the clinical isolates, the times to an ESP II result of susceptible (means +/- standard errors of the means) were 8.47 +/- 0.12 days (range, 7 to 10 days) and 8.73 +/- 0.29 days (range, 5 to 11 days) when the inoculum was prepared from a McFarland equivalent and from a seed bottle, respectively. The time to an ESP II result of resistant varied by drug and method of inoculum preparation, ranging from 5.50 +/- 0.22 days for ethambutol with the inoculum prepared from a McFarland standard to 8. 0 days for ethambutol with the inoculum prepared from a seed bottle. These data suggest that the ESP II system is a rapid and reliable method for testing susceptibilities of M. tuberculosis isolates to isoniazid and rifampin. Performance, however, may be suboptimal for ethambutol and streptomycin. Testing additional ethambutol-resistant and streptomycin-resistant strains with two concentrations of both drugs is necessary. PMID- 9738048 TI - Molecular epidemiology of penicillin-resistant Streptococcus pneumoniae isolates recovered in Italy from 1993 to 1996. AB - Thirty-nine penicillin-resistant Streptococcus pneumoniae isolates recovered among the approximately 700 pneumococcal strains collected from 1993 to 1996 in central and northern Italy were analyzed for several characteristics, including serotype, antibiotic susceptibility profile, chromosomal relatedness (by using pulsed-field gel electrophoresis [PFGE]), restriction fragment length polymorphism (RFLP) of the penicillin-binding protein (PBP) genes 1A, 2X, and 2B, and the presence of a variety of antibiotic resistance genes (determined by hybridization with appropriate DNA probes). The MICs of penicillin for most of the isolates (30 of 39) were high, in the range of 1 microgram/ml or higher, and these 30 isolates carried additional resistance traits to two or more drugs (erythromycin, chloramphenicol, co-trimoxazole, and tetracycline) and expressed serotypes 9, 19, and 23 and three distinct PFGE patterns. More than half (22 of 30) of the isolates for which MICs were high were identified as representatives of two widespread international epidemic clones of S. pneumoniae. The first one of these clones (seven isolates) expressed serotype 23F and possessed all properties characteristic of the widespread Spanish/USA international clone. Seven additional strains with serotype 19 also had the same PFGE pattern, PBP gene, and RFLP polymorphisms, and other properties typical of the serotype 23 Spanish/USA clone, suggesting that these strains were the products of a capsular transformation event (from serotype 23F to serotype 19) in which the Spanish/USA clone was the recipient. The second international clone was represented by eight serotype 9 isolates which were resistant to penicillin and trimethoprim sulfamethoxazole and had the molecular properties of the French/Spanish epidemic clone. The remaining eight isolates for which penicillin MICs were high appeared to represent a hitherto-undescribed "Italian" clone; they had a novel PFGE type, unique RFLPs for the PBP genes, and resistance to tetracycline, trimethoprim sulfamethoxazole, and erythromycin, and the penicillin MICs for these isolates were 2 to 4 microgram/ml. PMID- 9738049 TI - Comparison of In vitro activities of the new triazole SCH56592 and the echinocandins MK-0991 (L-743,872) and LY303366 against opportunistic filamentous and dimorphic fungi and yeasts. AB - The in vitro antifungal activities of SCH56592, MK-0991, and LY303366 against 83 isolates of Acremonium strictum, Aspergillus flavus, Aspergillus fumigatus, Aspergillus terreus, Bipolaris spp., Blastomyces dermatitidis, Cladophialophora bantiana, Fusarium oxysporum, Fusarium solani, Histoplasma capsulatum, Phialophora spp., Pseudallescheria boydii, Rhizopus arrhizus, Scedosporium prolificans, and Sporothrix schenckii were compared. The in vitro activities of these agents against 104 isolates of yeast pathogens of Candida spp., Cryptococcus neoformans, and Trichosporon beigelii were also compared. MICs were determined by following a procedure under evaluation by the National Committee for Clinical Laboratory Standards (NCCLS) for broth microdilution testing of the filamentous fungi (visual MICs) and the NCCLS M27-A broth microdilution method for yeasts (both visual and turbidimetric MICs). The in vitro fungicidal activity of SCH56592 was superior (minimum fungicidal concentrations [MFCs], 0.25 to 4 microgram/ml for 7 of 18 species tested) to those of MK-0991 and LY303366 (MFCs, 8 to >16 microgram/ml for all species tested) for the molds tested, but the echinocandins had a broader spectrum of fungicidal activity (MFCs at which 90% of strains are inhibited [MFC90s], 0.5 to 4 microgram/ml for 6 of 9 species tested) than SCH56592 (MFC90s, 0.25 to 8 microgram/ml for 4 of 9 species tested) against most of the yeasts tested. Neither echinocandin had in vitro activity (MICs, >16 microgram/ml) against C. neoformans and T. beigelii, while the SCH56592 MICs ranged from 0.12 to 1.0 microgram/ml for these two species. The MICs of the three agents for the other species ranged from <0.03 to 4 microgram/ml. These results suggest that these new agents have broad-spectrum activities in vitro; their effectiveness in the treatment of human mycoses is to be determined. PMID- 9738050 TI - Comparison of restriction enzyme analysis, arbitrarily primed PCR, and protein profile analysis typing for epidemiologic investigation of an ongoing Clostridium difficile outbreak. AB - During an outbreak of diarrhea in a general hospital in 1992, 166 Clostridium difficile isolates from 102 patients were typed by restriction enzyme analysis (REA), arbitrarily primed PCR (AP-PCR), and protein profile analysis (PP) techniques. A total of 18 types and 5 subtypes were identified by REA, 32 types were identified by AP-PCR, and 9 types were identified by PP. Analysis of the data indicated the presence of a predominant strain among 76, 75, and 84% of the isolates by REA, AP-PCR, and PP, respectively. Subsequently, 45 C. difficile isolates which had been collected in 1990 from 33 patients in the same hospital following a significant increase in the number of cases of diarrhea caused by C. difficile were studied by REA, AP-PCR, and PP typing techniques. Thirteen types and one subtype were identified by REA, 12 types were identified by AP-PCR, and 5 types were identified by PP. As with the isolates from 1992, a dominant strain was identified. This strain was represented by 53, 64, and 70% of the total number of isolates when the strains were typed by REA, AP-PCR, and PP, respectively. Every isolate (210 of 211) from both 1990 and 1992 that was available for typing was typeable by all three methods. Furthermore, the same dominant strain was identified in both 1990 and 1992 by each method. This study demonstrates that each of the three typing methods can be useful in epidemiologic investigations of C. difficile outbreaks and that one strain can be dominant in an institution over a number of years. PMID- 9738051 TI - Ultrasensitive reverse transcription-PCR assay for quantitation of human immunodeficiency virus type 1 RNA in plasma. AB - With the recent introduction of combination therapy, human immunodeficiency virus type 1 (HIV-1) RNA levels in plasma have been dramatically reduced, frequently to below the limit of quantitation (400 copies/ml of plasma) of the AMPLICOR HIV-1 MONITOR Test (Roche Diagnostic Systems). To achieve enhanced sensitivity of the AMPLICOR HIV-1 MONITOR Test, a modified specimen preparation procedure that allows input of RNA from 10-fold more plasma per amplification reaction was developed. This "ultrasensitive" method allows the accurate quantitation of plasma HIV-1 RNA levels as low as 50 copies/ml. A precision study yielded average within-run and between-run coefficients of variation (CV) of 24.8 and 9.6%, respectively. A multicenter reproducibility study demonstrated that the laboratory-to-laboratory reproducibility of this assay is good, with an average CV of 32%. The linear range of this test is between 50 and 50,000 copies/ml of plasma. RNA concentrations measured by the ultrasensitive and standard HIV-1 MONITOR tests exhibited good agreement within the shared linear range of the two methods. The two measurements were within a factor of 2 for 91% of the specimens tested, with the concentration measured by the ultrasensitive method being only slightly lower (median, 22% lower). Preliminary studies suggest that this assay will prove to be useful for predicting the stability of viral suppression in patients whose RNA levels drop below 400 copies/ml in response to highly active antiretroviral therapy. PMID- 9738052 TI - Field validation of laboratory tests for clinical diagnosis of sheep-associated malignant catarrhal fever. AB - Until recently, sheep-associated malignant catarrhal fever (SA-MCF) was diagnosed mainly on the basis of clinical presentation and histopathological changes. Using clinically diagnosed field cases, we have evaluated a seminested PCR and a competitive inhibition enzyme-linked immunosorbent assay (CI-ELISA) and compared these assays in the diagnosis of SA-MCF in cattle with histopathology as a provisional "gold standard." Samples from 44 cattle with clinical signs suggestive of SA-MCF were examined by histopathology, PCR, and CI-ELISA. In addition, samples from healthy cattle were evaluated by PCR (n = 96) and CI-ELISA (n = 75). Based on histopathology, 38 of the 44 clinical cases were classified as SA-MCF positive, 3 were classified as inconclusive, and 3 were classified as SA MCF negative. The sensitivity of PCR was 95 to 97%, whereas the specificity ranged between 94 and 100%. The CI-ELISA showed a sensitivity of 56 to 87% and a specificity between 91 and 100%. In the field, there is good correlation between the diagnoses of SA-MCF by histopathology, PCR, and CI-ELISA. These data also confirm the close association of ovine herpesvirus 2 with SA-MCF in Switzerland. PMID- 9738053 TI - Comparison of different DNA fingerprinting techniques for molecular typing of Bartonella henselae isolates. AB - Seventeen isolates of Bartonella henselae from the region of Freiburg, Germany, obtained from blood cultures of domestic cats, were examined for their genetic heterogeneity. On the basis of different DNA fingerprinting methods, including pulsed-field gel electrophoresis (PFGE), enterobacterial repetitive intergenic consensus (ERIC)-PCR, repetitive extragenic palindromic (REP) PCR, and arbitrarily primed (AP)-PCR, three different variants were identified among the isolates (variants I to III). Variant I included 6 strains, variant II included 10 strains, and variant III included only one strain. By all methods used, the isolates could be clearly distinguished from the type strain, Houston-1, which was designated variant IV. A previously published type-specific amplification of 16S rDNA differentiated two types of the B. henselae isolates (16S rRNA types 1 and 2). The majority of the isolates (16 of 17), including all variants I and II, were 16S rRNA type 2. Only one isolate (variant III) and the Houston-1 strain (variant IV) comprised the 16S rRNA type 1. Comparison of the 16S rDNA sequences from one representative strain from each of the three variants (I to III) confirmed the results obtained by 16S rRNA type-specific PCR. The sequences from variant I and variant II were identical, whereas the sequence of variant III differed in three positions. All methods applied in this study allowed subtyping of the isolates. PFGE and ERIC-PCR provided the highest discriminatory potential for subtyping B. henselae strains, whereas AP-PCR with the M13 primer showed a very clear differentiation between the four variants. Our results suggest that the genetic heterogeneity of B. henselae strains is high. The methods applied were found useful for typing B. henselae isolates, providing tools for epidemiological and clinical follow-up studies. PMID- 9738055 TI - Multiplex PCR for typing and subtyping influenza and respiratory syncytial viruses. AB - A multiplex reverse transcription (RT)-PCR method that has been developed is capable of detecting and subtyping influenza A (H1N1 and H3N2) and B viruses as well as respiratory syncytial virus (RSV) types A and B in respiratory clinical samples taken as part of a national community-based surveillance program of influenza-like illness in England and Wales. The detection of each different pathogen depended on distinguishing five amplification products of different sizes on agarose gels following RT-PCR with multiple primer sets. The multiplex RT-PCR was tested with 65 nasopharyngeal apirates from which RSV had been isolated and 237 combined nose and throat swabs from which influenza A (H1N1 and H3N2) or B virus had been detected by virus isolation, as well as 40 respiratory samples from which other viruses including cytomegalovirus, herpes simplex virus, enteroviruses, and parainfluenza viruses had been grown. For the typing and subtyping of influenza A and B viruses and RSV types A and B, the multiplex RT PCR gave an excellent (100%) correlation with the results of conventional typing and subtyping with specific antisera. Multiplex RT-PCR can also be used to accurately detect more than one viral template in the same reaction mixture, allowing viral coinfections to be identified with the same respiratory specimen. PMID- 9738054 TI - Assessment of hepatitis C virus sequence complexity by electrophoretic mobilities of both single-and double-stranded DNAs. AB - To assess genetic variation in hepatitis C virus (HCV) sequences accurately, we optimized a method for identifying distinct viral clones without determining the nucleotide sequence of each clone. Twelve serum samples were obtained from seven individuals soon after they acquired HCV during a prospective study, and a 452-bp fragment from the E2 region was amplified by reverse transcriptase PCR and cloned. Thirty-three cloned cDNAs representing each specimen were assessed by a method that combined heteroduplex analysis (HDA) and a single-stranded conformational polymorphism (SSCP) method to determine the number of clonotypes (electrophoretically indistinguishable cloned cDNAs) as a measure of genetic complexity (this combined method is referred to herein as the HDA+SSCP method). We calculated Shannon entropy, incorporating the number and distribution of clonotypes into a single quantifier of complexity. These measures were evaluated for their correlation with nucleotide sequence diversity. Blinded analysis revealed that the sensitivity (ability to detect variants) and specificity (avoidance of false detection) of the HDA+SSCP method were very high. The genetic distance (mean +/- standard deviation) between indistinguishable cloned cDNAs (intraclonotype diversity) was 0.6% +/- 0.9%, and 98.7% of cDNAs differed by <2%, while the mean distance between cloned cDNAs with different patterns was 4.0% +/- 3.2%. The sensitivity of the HDA+SSCP method compared favorably with either HDA or the SSCP method alone, which resulted in intraclonotype diversities of 1.6% +/ 1.8% and 3.5% +/- 3.4%, respectively. The number of clonotypes correlated strongly with genetic diversity (R2, 0.93), but this correlation fell off sharply when fewer clones were assessed. This HDA+SSCP method accurately reflected nucleotide sequence diversity among a large number of viral cDNA clones, which should enhance analyses to determine the effects of viral diversity on HCV associated disease. If sequence diversity becomes recognized as an important parameter for staging or monitoring of HCV infection, this method should be practical enough for use in laboratories that perform nucleic acid testing. PMID- 9738056 TI - Use of molecular and reference susceptibility testing methods in a multicenter evaluation of MicroScan dried overnight gram-positive MIC panels for detection of vancomycin and high-level aminoglycoside resistances in enterococci. AB - Modified MicroScan gram-positive MIC no. 8 panels (PM-8) were analyzed for their improved ability to detect vancomycin resistance (VR) and high-level aminoglycoside resistance (HLAR) in enterococci. A validation study design that utilized selected challenge strains, recent clinical isolates, and reproducibility experiments in a multicenter format was selected. Three independent medical centers compared the commercial panels to reference broth microdilution panels (RBM) and Synergy Quad Agar (QA). Resistance was verified by demonstration of VR and HLAR genes by PCR tests. The study was conducted in three phases. (i) In the challenge phase (CP), two well-characterized sets of enterococci were obtained from the Centers for Disease Control and Prevention; one set contained 50 isolates for VR testing and one contained 48 isolates for HLAR testing. In addition, a set of 47 well-characterized isolates representing diverse geographic areas, obtained from earlier national surveillance studies, was tested at the University of Iowa College of Medicine (UICM). (ii) In the efficacy phase (EP), each laboratory tested 50 recent, unique clinical isolates by all methods. (iii) In the reproducibility Phase (RP), each laboratory tested the same 10 strains by all methods in triplicate on three separate days. All isolates from the EP were sent to the UICM for molecular characterization of vanA, -B, -C1, -C2-3, and HLAR genes. In the CP, the ranking of test methods by error rates (in parentheses; very major and major errors combined, versus PCR results) were as follows: for high-level streptomycin resistance (HLSR), QA (12.0%) > PM-8 (5.2%) > RBM (1.6%); for high-level gentamicin resistance (HLGR), RBM (3.7%) > PM-8 (3.1%) > QA (2.6%); and for VR, RBM = QA (3.0%) > PM-8 (1.2%). In the EP, agreement between all methods and the reference PCR result was 98.0% for HLSR, 99.3% for HLGR, and 98. 6% for VR. In the RP, the percentages of results +/- 1 log2 dilution of the all-participant mode were as follows: for VR, 100% (PM-8), 98.9% (QA), and 90.0% (RBM); for HLSR, 99.6% (RBM), 98.5% (PM-8), and 82.2% (QA); and for HLGR, 99.6% (RBM), 99.3% (PM-8), and 98.1% (QA). The ability of the PM-8 to detect VR and HLAR in enterococci was comparable to those for reference susceptibility and molecular PCR methods and was considered acceptable for routine clinical laboratory use. PMID- 9738057 TI - Hepatitis C virus serotype-specific core and NS4 antibodies in injecting drug users participating in the Amsterdam cohort studies. AB - In the present study, the RIBA HCV serotyping SIA was evaluated with a cohort of injecting drug users. Serotyping may be a rapid and cost-effective method of determining genotypes in cohort studies. In this study, hepatitis C virus (HCV) antibody-positive sera from a cohort of 331 chronically infected injecting drug users, of which 167 were coinfected with human immunodeficiency virus (HIV), were serotyped by the RIBA HCV Serotyping SIA. Among the 331 specimens, serotype specific antibodies were detected in 250 (sensitivity, 75. 5%), in which serotype 1 was predominant (57.2%), followed by serotype 3 (26.8%). Among the 331 specimens, 164 were HIV negative, and serotype-specific antibodies were detected in 151 (sensitivity, 92.1%), in which serotype 1 was predominant (59.6%), followed by serotype 3 (33.8%). For a subset of 58 samples taken from 19 chronically infected HCV seroconverters with a mean follow-up of 5 years, serotypes were compared with genotypes, which were determined by a line probe assay (HCV LiPa) and by direct sequencing of the products obtained by nested PCR of the 5' untranslated region. Among the 58 samples with known genotypes, serotype-specific antibodies were detected in 38 (total sensitivity, 65.5%), with a specificity of 78.9%. Thirty of these serotypeable samples revealed a serotype that corresponded to the genotype in the 58 samples (total positive predictive value, 51.7%). Of the 58 samples, 23 were coinfected with HIV, and when these were excluded, the total sensitivity increased to 76.5%, with a total specificity of 80.8% and a total positive predictive value of 61.8%. The serotyping assay showed a high total sensitivity (96.3%) for samples positive by HCV RIBA, version 3.0, with four bands. We conclude that the sensitivity of the RIBA HCV serotyping SIA is limited by the immunocompetence of the HCV-infected host. In general, samples from HIV-negative individuals containing genotype 1a had higher sensitivity, specificity, and concordance in the serotyping assay compared with genotyping, whereas samples containing genotype 3a were found to be more cross reactive and untypeable. Therefore, the prevalence of genotypes other than genotype 1 could be underestimated if they are determined by serotyping, and improvements in specificity are recommended. PMID- 9738058 TI - Detection of Candida dubliniensis in oropharyngeal samples from human immunodeficiency virus-infected patients in North America by primary CHROMagar candida screening and susceptibility testing of isolates. AB - Candida dubliniensis has been associated with oropharyngeal candidiasis in patients infected with human immunodeficiency virus (HIV). C. dubliniensis isolates may have been improperly characterized as atypical Candida albicans due to the phenotypic similarity between the two species. Prospective screening of oral rinses from 63 HIV-infected patients detected atypical dark green isolates on CHROMagar Candida compared to typical C. albicans isolates, which are light green. Forty-eight atypical isolates and three control strains were characterized by germ tube formation, differential growth at 37, 42, and 45 degreesC, identification by API 20C, fluorescence, chlamydoconidium production, and fingerprinting by Ca3 probe DNA hybridization patterns. All isolates were germ tube positive. Very poor or no growth occurred at 42 degreesC with 22 of 51 isolates. All 22 poorly growing isolates at 42 degreesC and one isolate with growth at 42 degreesC showed weak hybridization of the Ca3 probe with genomic DNA, consistent with C. dubliniensis identification. No C. dubliniensis isolate but only 18 of 28 C. albicans isolates grew at 45 degreesC. Other phenotypic or morphologic tests were less reliable in differentiating C. dubliniensis from C. albicans. Antifungal susceptibility testing showed fluconazole MICs ranging from /= 50 years of age; Group B, 44 patients < 50. Using Osserman's classification and the duration of the disease prior to hospitalization, we observed no significant differences between the two groups although thymoma were more prevalent in Group A. All 58 patients underwent extended thymectomy, with no surgical mortalities. The mean duration of tracheal intubation after thymectomy for Group A was 6.25 days; for Group B, 4.66 days without statistically significant differences between both groups. Using Masaoka's criteria, we evaluated the clinical course of myasthenia gravis following extended thymectomy for each of the 58 patients. The remission rates in Groups A and B were 28.6% and 29.5%, respectively; the improvement rates, 71.4% and 79.5%, respectively with no significant differences among groups. These findings suggest that the clinical course of myasthenia gravis following extended thymectomy is not age-dependent and that extended thymectomy is a clinically safe and effective treatment option for myasthenia gravis patients >/= 50 years of age. PMID- 9738120 TI - Thymectomy for myasthenia gravis: outcome of treatment in a tertiary hospital. AB - The clinical records of 24 of the 31 patients who underwent thymectomy for myasthenia gravis from January 1991 to December 1995 were reviewed. The mean age of patients was 36.5 years, with a male : female ratio of 1 : 5. The average duration of symptoms prior to consultation was 16.6 months. Most of the patients presented diplopia with clinical stage IIA prior to surgery. The most common procedure utilized was thymectomy via a median sternotomy under general anesthesia-epidural anesthesia. Five of the 24 patients (21%) were immediately extubated postoperatively while 19 remained intubated, with 12 patients attached to a ventilator. Extubation ranged from the first postoperative day up to more than two weeks postoperatively. The average length of ICU stay was 8.1 days and the average length of hospital stay was 31.1 days. The most common morbidity was pneumonia (38%). One patient had a brief reactive psychosis and another patient had phrenic nerve injury which manifested 2 weeks postoperatively. The average length of follow-up was 14.8 months and the mean follow-up period was 24.5 months. There was a general trend of decrease in the dose of medication. However, there was an increase in the medication in 3 patients with 2 showing clinical improvement. PMID- 9738121 TI - Efficacy of a left atrial procedure for treatment of chronic atrial fibrillation in elderly patients. AB - A simple left atrial procedure for atrial fibrillation (AF) was performed in 16 elderly patients (age over 70 years old) with chronic AF associated with mitral and/or aortic valvular diseases. Chronic AF was eliminated upon discharge in 13 out of the 16 patients (81%). Twelve of the 13 patients (92%) whose AF had disappeared recovered the atrial kick of their right atrium, and 9 patients (70%) recovered the atrial kick of their left atrium. A simple surgical procedure to the left atrium was effective in the treatment of chronic AF associated with mitral valve disease in elderly patients. This simple procedure is preferable to other methods for the elimination of chronic AF with mitral valve disease in elderly patients. PMID- 9738122 TI - Minimally invasive left anterior descending coronary artery revascularisation in high-risk patients with three-vessel disease. AB - We reviewed our experience over a 12 month period with using minimally invasive direct coronary artery bypass (MIDCAB) for the management of high-risk patients with three-vessel coronary artery disease. Twenty patients (4 females, mean age 67 years) received left internal mammary artery (LIMA) grafts to the left anterior descending (LAD) coronary artery. Associated co-morbidity included: severe chronic renal failure, severe extensive arteriopathy, chronic obstructive airway disease, poor general condition and severely impaired left ventricular function. There was one early postoperative mortality and no other cardiac related morbidity. Graft patency investigated, using angiography was 90%, and 5 patients underwent follow-up angioplasty to other coronary arteries. All patients remain entirely angina free at a follow-up period between 3 and 12 months. We conclude that MIDCAB is a safe and effective approach for managing high-risk patients with three-vessel coronary artery disease. PMID- 9738123 TI - The role of aggressive medical therapy along with early surgical intervention in the cure of Brucella endocarditis. AB - Timing of surgical intervention in Brucella endocarditis remains controversial. In this report, we review our experience in an attempt to collect some information on the best approach for this entity. From June 1992 to December 1996, 5 male patients between the ages of 20 and 35 years with Brucella endocarditis were operated on in our centre. Three of them had native valve endocarditis (NVE) and 2 with prosthetic valve endocarditis (PVE). All patients belonged to New York Heart Association (NYHA) class III-IV. All had developed anti Brucella antibodies with serum agglutination titres of > 320 and the sera tested from 3 patients were Enzyme Linked Immunosorbent Assay (ELISA) positive for anti-Brucella IgM and/or IgG antibodies. In 3 cases 2D-echocardiography showed large vegetation on the affected valve. Blood cultures were positive in 4 patients, 2 of them (one each of NVE and PVE) had the valve material culture positive for Brucella. All cases were treated with a combination of doxycycline, refampicine and gentamicin before surgery. Major indication for surgical intervention was severe haemodynamic instability which developed during the course of antibiotic therapy either early (3 cases) or late (2 cases). All patients became asymptomatic at the end of 7 days postoperatively. On the follow up for a period of 8-51 months, all patients were in NYHA class I-II without evidence of recurrence of infection. These data suggest that in either NVE or PVE Brucella, medical therapy alone may not be sufficient due to the eventual haemodynamic deterioration secondary to valve tissue destruction or dysfunction of the prosthetic valve caused by the infective process. Therefore, a combination of aggressive medical therapy with multiple bactericidal antibiotics and early surgical intervention may result in a successful outcome, but further studies are needed to reach a reliable conclusion. PMID- 9738124 TI - A new preshaped catheter for right ventriculography with minimal artificial tricuspid regurgitation. AB - A new type of preshaped angiographic catheter for right ventriculography to be inserted from the femoral vein has been developed. Right ventriculography using this catheter was conducted in about five hundred clinical cases. The result has revealed that this catheter is very useful in clinical settings without causing catheter-induced tricuspid regurgitation (TR) allowing easier and safer right ventriculography as compared with conventional catheters. PMID- 9738125 TI - Thromboembolism leading to myocardial ischaemia in a patient requiring a fenestrated Fontan operation. AB - We report a case of air/thromboembolism of the left anterior descending coronary artery complicating a fenestrated Fontan procedure treated with aprotinin. The patient was successfully treated by saphenous vein grafting to the left anterior descending coronary artery and reconversion of the Fontan circulation to the original cavo-pulmonary connection. PMID- 9738126 TI - Cryopreserved ascending aortic homograft for Stanford B dissecting aneurysm. AB - A 68 year-old woman underwent replacement of the descending thoracic aorta with a cryopreserved aortic homograft for the treatment of a Stanford B dissecting aneurysm. After surgery, the patient made an uneventful recovery without recurrent infection. In such cases, the use of aortic homografts to replace aortic segments is a promising and effective therapeutic option. PMID- 9738127 TI - Bentall operation, total aortic replacement and mitral valve replacement for a young adult with Marfan syndrome: a case of three-staged operation. AB - In Marfan syndrome, the most common cardiovascular abnormalities are dilatation of the aorta and aortic valve regurgitation in adult patients. Mitral valve dysfunction is the most common cause of morbidity and mortality in infants and children with Marfan syndrome, and is not frequently operated on in adult Marfan patients who undergo surgery for diseases of the aortic root and total aorta. This report describes a successfully three-staged operation for a 24 year-old man with Marfan syndrome who underwent an emergent Bentall operation and aortic arch replacement, total aortic replacement and mitral valve replacement over 2 years. Mitral valve regurgitation was mild but increased after the second operation. The graft was tightly adhesive and invasive to the sternum. Endoscopic view was helpful to avoid graft damage at resternotomy. The postoperative course was uneventful in each operation. Microscopic examination of the mitral valve leaflets showed abnormal increase of mucopolysaccharides, and disruption and fragmentation of elastic fibers. PMID- 9738128 TI - A case successfully treated by conservative management for mediastinitis and infected composite graft due to methicillin-resistant coagulase negative staphylococcus. AB - A 72 year-old man underwent a Bentall procedure for aortic regurgitation secondary to annulo-aortic ectasia and ascending aortic aneurysm. On the 11th postoperative day, the C-reactive protein (CRP) level and white blood cell (WBC) count rose. Echocardiography and a computed tomographic scan showed the appearance of pericardial effusion. A diagnosis of mediastinitis and composite graft infection was made, and mediastinal drainage and irrigation were performed. Methicillin-resistant coagulase negative staphylococcus (MRCNS) was identified as the causative organism. Vancomycin, arbekacin and minocycline were used intravenously. Additionally, a continuous mediastinal irrigation was performed through the chest tubes. CRP level and WBC count were gradually reduced to normal range. He has now been free from signs of infection for more than 3 years. Because MRCNS is considered less virulent than methicillin-resistant Staphylococcus aureus, mediastinitis and composite graft infection due to MRCNS might be treatable by such conservative therapy even in patients with prosthetic implants. Since MRCNS often becomes ubiquitous, preventing infections by strict attention to asepsis is important. PMID- 9738129 TI - Effects of split exercise sessions on excess postexercise oxygen consumption and resting metabolic rate. AB - This study involved examining how splitting a 30-min exercise bout on a cycle ergometer into two equal sessions affects excess postexercise oxygen consumption (EPOC) and resting metabolic rate (RMR). In this study, 10 male volunteers (age = 23+/-3.8) participated in two exercise trials, which were randomly assigned in a counterbalanced design and separated by 40 hr. One trial was 30 min of exercise at 70% VO(2)max (CONT), followed by a 40-min measurement of EPOC. The second trial was divided into two 15-min sessions (SPLIT), separated by 6 hr. A 20-min measurement of EPOC followed each SPLIT session. Results indicated that the combined magnitude of EPOCs from SPLIT (7,410+/-1,851 ml) was significantly greater than that from CONT (5,278+/-1305 ml). Data indicate that dividing a 30 min exercise session in to two parts for these individuals significantly increases magnitude of EPOC but does not affect RMR. PMID- 9738130 TI - Association between aerobic capacity and carotid-cardiac baroreflex responsiveness in women. AB - The purpose of this study was to determine whether the positive correlation between carotid-cardiac baroreflex responsiveness and aerobic capacity (VO(2)max) that has been reported in men also occurs in women. Carotid-cardiac baroreflex responsiveness was tested in 40 healthy, normotensive women (age 18-35) using the variable neck pressure technique. Participants were subdivided into endurance trained (ET; n = 11) and untrained (UT; n = 9) groups. No significant between group difference was found in the range or gain of the carotid-cardiac baroreflex response despite a lower resting HR in the ET group. When participants were subdivided into high (HI; n = 13) and low (LO; n = 17) responders based on reflex RRI responses to CTP changes, no significant between-group differences were found in resting HR or VO(2)max levels. It was concluded that aerobic capacity (VO(2)max) is not a good predictor of cardiac-carotid baroreflex responsiveness in healthy women. PMID- 9738131 TI - Influence of diet and/or exercise on body composition and cardiorespiratory fitness in obese women. AB - The purpose of this study was to measure the influence of diet, exercise, or both on body composition and cardiorespiratory fitness in obese women. Ninety-one obese subjects were randomized into one of four groups: diet (D) (4.19-5.44 MJ or 1,200-1,300 kcal/day), exercise (E) (five 45-min sessions at 78.5+/-0.5% maximum heart rate), exercise and diet (ED), and controls (C). Maximal aerobic power and body composition were measured in all subjects before and after a 12-week diet intervention period. Subjects in D and ED lost 7.8+/-0.7 and 8.1+/-0.6 kg body mass, with no significant change for E relative to C. Losses of percent body fat and fat mass were significantly greater in D and ED but not in E relative to C. The change in VO2max was greater in ED and E but not D when compared to C. Results indicate that moderate aerobic exercise training during a 12-week period has no discernible effects on body composition but does improve cardiorespiratory fitness in dieting obese women. PMID- 9738132 TI - Serum ferritin and anemia in trained female athletes. AB - The aim of this study was to establish whether extremely low serum ferritin values in female athletes were associated with indications of iron deficiency anemia and whether serum ferritin values were influenced by the type of training or participants' body size. Hematological data collected during 6 years at the Australian Institute of Sport were reviewed to quantify changes in serum ferritin concentration associated with training and to establish whether decrements in serum ferritin were associated with any change in hemoglobin concentration, mean corpuscular volume, or mean corpuscular hemoglobin concentration. Mean serum ferritin concentrations of 7.5 microg x L(-1) were not associated with any indication of iron-deficiency anemia. Serum ferritin declined by approximately 25% with the onset of rigorous daily training (p < .01) whether training was predominantly weight-bearing or non-weight-bearing. Rowers had significantly higher ferritin concentrations than basketball players of similar stature (p=.02). We conclude that considerable background information such as the stage of training, specific sport, and previous blood results should be sought when interpreting serum ferritin concentrations in female athletes. PMID- 9738134 TI - Pyruvate: beyond the marketing hype. PMID- 9738133 TI - Dietary and performance assessment of elite soccer players during a period of intense training. AB - This study examined the nutritional and performance status of elite soccer players during intense training. Eight male players (age 17+/-2 years) of the Puerto Rican Olympic Team recorded daily activities and food intake over 12 days. Daily energy expenditure was 3,833+/-571 (SD) kcal, and energy intake was 3,952+/ 1,071 kcal, of which 53.2+/-6.2% (8.3 g x kg BW(-1)) was from carbohydrates (CHO), 32.4+/-4.0% from fat, and 14.4+/-2.3% from protein. With the exception of calcium, all micronutrients examined were in accordance with dietary guidelines. Body fat was 7.6+/-1.1% of body weight. Time to completion of three runs of the soccer-specific test was 37.65+/-0.62 s, and peak torques of the knee flexors and extensors at 60 degrees x s(-1) were 139+/-6 and 225+/-9 N x m, respectively. Players' absolute amounts of CHO seemed to be above the minimum recommended intake to maximize glycogen storage, but calcium intakes were below recommended. Their body fat was unremarkable, and they had a comparatively good capacity to endure repeated bouts of intense soccer-specific exercise and to exert force with their knee extensors and flexors. PMID- 9738135 TI - Nutrition, physical activity, and bone health in women. AB - Calcium and vitamin D can significantly impact bone mineral and fracture risk in women. Unfortunately, calcium intakes in women are low and many elderly have poor vitamin D status. Supplementation with calcium (approximately 1000 mg) can reduce bone loss in premenopausal and late postmenopausal women, especially at sites that have a high cortical bone composition. Vitamin D supplementation slows bone loss and reduces fracture rates in late postmenopausal women. While an excess of nutrients such as sodium and protein potentially affect bone mineral through increased calcium excretion, phytoestrogens in soy foods may attenuate bone loss through estrogenlike activity. Weight-bearing physical activity may reduce the risk of osteoporosis in women by augmenting bone mineral during the early adult years and reducing the loss of bone following menopause. High-load activities, such as resistance training, appear to provide the best stimulus for enhancing bone mineral; however, repetitive activities, such as walking, may have a positive impact on bone mineral when performed at higher intensities. Irrespective of changes in bone mineral, physical activities that improve muscular strength, endurance, and balance may reduce fracture risk by reducing the risk of falling. The combined effect of physical activity and calcium supplementation on bone mineral needs further investigation. PMID- 9738136 TI - Practical body composition assessment for children, adults, and older adults. AB - This paper provides an overview of practical methods for assessing body composition of children, adults, and older adults. Three methods commonly used in field and clinical settings are skinfolds, bioelectrical impedance analysis, and anthropometry. For each method, standardized testing procedures, sources of measurement error, recommendations for technicians, and selected prediction equations for each age category are presented. The skinfold method is appropriate for estimating body fat of children (6-17 years) and body density of adults (18 60 years) from diverse ethnic groups. Likewise, bioimpedance is well suited for estimating the fat-free mass of children (10-19 years) as well as American Indian, black, Hispanic, and white adults. Anthropometric prediction equations that use a combination of circumferences and bony diameters are recommended for older adults (up to 79 years of age), as well as obese men and women. PMID- 9738137 TI - Effective nutrition support programs for college athletes. AB - This paper presents an overview of the Husky Sport Nutrition Program at the University of Washington. This program is a component of the Department of Intercollegiate Athletics Total Student-Athlete Program, an NCAA-sponsored CHAMPS/Life Skills Program that provides life skills assistance to student athletes. Successful integration of a sport nutrition program requires an understanding of the athletic culture, physiological milestones, and life stressors faced by college athletes. The sport nutritionist functions as an educator, counselor, and administrator. Team presentations and individual nutrition counseling provide athletes with accurate information on healthy eating behaviors for optimal performance. For women's sports, a multidisciplinary team including the sport nutritionist, team physician, clinical psychologist, and athletic trainer work to prevent and treat eating disorders. Case studies are presented illustrating the breadth of nutrition-related issues faced by a sport nutritionist working with college athletes. PMID- 9738138 TI - Mergers. They're still smokin'. PMID- 9738140 TI - Risk management. Guideline payoff. PMID- 9738139 TI - Payment. Quick buck. PMID- 9738141 TI - A county cares for its own. Public-private venture in Florida's Duval County struggles hard to serve uninsured kids. PMID- 9738142 TI - Leading the way. 1998 leadership survey. AB - Compliance and marketing are hot, info tech gets low marks, and quality is a bone contention. Here's what more than 700 hospital, physician, and health plan leaders told Hospitals & Health Networks and MGMA via the 1998 Leadership Survey. PMID- 9738143 TI - Hospital heavies. Venture capital bulks up companies that outsource medicine's newest specialty: inpatient-only care. AB - They're the designated drivers of inpatient care, cutting hospital stays by 19 percent on average. Yet as venture capital firms infuse hospitalist startup companies, some primary care doctors complain that their sickest patients are being taken away from them. PMID- 9738144 TI - Prospective payment gets legs. AB - Any change seems like an improvement when it comes to Medicare's current payment system for outpatient care. But a fixed-fee system is largely untested, and hospitals have plenty of questions. Here are some answers. PMID- 9738145 TI - Software for slimming down. AB - High-tech is showing where hospitals can trim expenses--from clinical operations to strategic planning and business development. But it's still up to you to make the hard decisions. PMID- 9738147 TI - HMO backlash. Doctors who just say no. PMID- 9738146 TI - Hollywood squares. PMID- 9738148 TI - War zone medicine. M*A*S*H meets managed care. PMID- 9738149 TI - Palliative care. Gain on pain. PMID- 9738151 TI - Conflicts of interest. Research gone bad? PMID- 9738150 TI - Insurance reform. Kentucky turnabout. PMID- 9738153 TI - Hospital pulse ... January 1998. PMID- 9738152 TI - Nurse shortage. Where have all the RNs gone? PMID- 9738154 TI - Stroke: a systems approach. PMID- 9738155 TI - The mind of the machine. PMID- 9738157 TI - RE: MJ Krahn, NM Pettigrew, TE Cuddy. Unusual side effects due to warfarin. 1998; 14:90-3. PMID- 9738156 TI - RE: K Zufelt, HC Rosenberg, MD Li, GI Joubert. The electrocardiogram and the secundum atrial septal defect: a reexamination in the era of echocardiography. 1998; 14:227-32. PMID- 9738158 TI - [Standards of pediatric echocardiography]. PMID- 9738159 TI - [Standards of pediatric electrophysiology]. PMID- 9738160 TI - [Standards of heart catheterization and interventional cardiology in pediatrics]. PMID- 9738161 TI - Ten-year trends of heart disease risk factors in the Halifax County MONICA population. MONItoring of trends and determinants in CArdiovascular disease. AB - OBJECTIVE: To investigate trends in heart disease risk factors (RFs) in the general population of Halifax County, Nova Scotia during a 10-year period. DESIGN: Two independent random samples of the population of Halifax County were surveyed in 1985 and 1995; age ranges were 25-64 years and 25-74 years. Blood pressure, cholesterol and body weight were measured. Smoking and health history were obtained by questionnaire. MAIN RESULTS: Participation rate was 66.3% in 1985 and 1995. All RFs were negatively correlated with education attainment. RF changes from 1985 to 1995 were related to education level. Among survey participants, mean body mass index increased from 26.7 kg/m2 to 27.6 kg/m2 (P + 0.005) for men, and from 25.5 kg/m2 to 27.3 kg/m2 (P < 0.00001) for women. Average smoking rate increased from 32.0% to 34.6% (not significant) in men and from 27.7% to 29.1% (not significant) in women. Age-specific smoking rate increased by 13% (P = 0.14) in younger women and decreased by 10% in older women. (P = 0.00). Mean levels of blood cholesterol decreased by 0.2 mmol/L (P = 0.002) in men and 0.1 mmol/L (P = 0.20) in women. Systolic blood pressure increased by 6.3 mmHg (P < 0.0001) in men and by 7.9 mmHg (P < 0.0001) in women, being steepest in the lower education group. Mortality predicted from RFs declined between the survey years, but less than the observed mortality. This discrepancy may result from the effect of medical care or the delayed effect of RF changes. CONCLUSIONS: Some risk factors show a disturbing trend, indicating that an increased effort or a change in strategy is needed to combat the risk of ischemic heart disease. PMID- 9738162 TI - Captopril and angiotensin II receptor antagonist therapy in a pacing model of heart failure. AB - Twenty-four splenectomized dogs were subjected to rapid right ventricular pacing (RRVP) at 250 beats/min for five weeks. During the final three weeks, four groups six dogs were untreated or treated with captopril alone, with the angiotensin II type 1 (AT1) receptor antagonist L158,809 alone or with the two drugs combined by constant intravenous infusion. Hemodynamic studies were carried out during light anesthesia at baseline, and after two and five weeks of pacing. Total vascular capacitance and stressed blood volume were calculated from the mean circulatory filling pressure during transient circulatory arrest after acetylcholine administration at three different circulating volumes. Central blood volume and cardiac output were measured by thermodilution. Severe heart failure was present in the untreated group after five weeks of RRVP, characterized by low cardiac output and total vascular capacitance, high right atrial and pulmonary capillary wedge and mean circulatory filling pressure, plus increased stressed and central blood volumes. While L158,809 had not effect, captopril alone or combined with L158,809 ameliorated the reduction in total vascular capacitance, and reduced right atrial and mean circulatory pressure and stressed blood volumes. Combined therapy reduced pulmonary capillary wedge pressure. Thus, angiotensin-converting enzyme inhibition with captopril was effective in this model of chronic low output heart failure, whereas AT1 receptor antagonism was not. PMID- 9738163 TI - Experimental branch pulmonary artery stenosis angioplasty using a novel cutting balloon. AB - To determined the safety and efficacy of a bladed balloon in the treatment of branch pulmonary artery stenosis, a model of left pulmonary artery stenosis was surgically created in two-week-old pigs. Seven pigs underwent angioplasty, five with the bladed balloon and two with conventional balloons. Overall, acute results showed a fall in the peak systolic pressure gradients from 8.3 +/- 2.3 mmHg to 3.2 +/- 3.1 mmHg and an increase in the minimum stenotic diameters from 4.5 +/- 2mm to 5.6 +/- mm. Acute pathological examination after cutting angioplasty showed regular luminal cuts that healed completely by four to six weeks in chronically surviving animals. Two of three surviving animals had persistent vessel enlargement at follow-up with one showing little overall change. Cutting balloons are effective in branch pulmonary artery angioplasty and may have clinical applications. PMID- 9738164 TI - Beta-blocker therapy for congestive heart failure: a systemic overview and critical appraisal of the published trials. AB - OBJECTIVE: To evaluate the effect of beta-blockers on mortality and morbidity, and to provide an appraisal of the reliability of the available data. DATA SOURCES: MEDLINE search for trials of beta-blockers for congestive heart failure (CHF). STUDY SELECTION: All randomized trials of beta-blockers versus placebo, or greater than one month's duration, in patients with CHF. Eighteen published trials involving 2986 patients were selected. DATA EXTRACTION: Independently by two authors. DATA SYNTHESIS: The Yusuf-Peto method for combining data was used. Data were available on mortality in 2841 patients (95%), on hospitalization for heart failure in 1514 (51%) and on heart transplantation in 2330 (79%). There was a lower rate of death in the active treatment group (131 of 1606) [8.2] versus 155 of 1235 [12.6%]; OR = 72; 99% CI 0.51 to 1.00), a lower rate of hospitalization for heart failure (137 of 756 [18.1%] versus 218 of 758 [28.7%]; OR = 0.54; 99% CI 0.39 to 0.74) and a trend towards a lower proportion of patients receiving heart transplantation (15 of 1354 [l.1%] versus 26 of 976 [2.7%]; OR = 0.45; 99% CI 0.20 to 1.03). Ventricular function improved; however, there was no effect on exercise duration. Although the effects on mortality were nominally statistically significant, the use of formal methods of interim monitoring adapted for meta-analyses suggests that substantially more patients still need to be studied in large scales trials to provide reliable and conclusive evidence. CONCLUSIONS: While the available data on the use of beta blockers in CHF appear to be promising, they are neither complete nor robust. The routine use of beta-blockers in patients with heart failure should wait the results of ongoing studies. PMID- 9738165 TI - The role of glycoprotein IIb/IIIa platelet inhibition in percutaneous coronary intervention in the stent era. AB - In autumn 1996, shortly after the platelet glycoprotein (GP) IIb/IIIa inhibitor abciximab was approved for clinical use by the Health Protection Branch of Health Canada, seven interventional cardiologists met in a roundtable forum to review the use of abciximab in percutaneous transluminal coronary angioplasty (PTCA). While a compelling body of data was presented that argued strongly for adjunctive abciximab in conventional balloon angioplasty, the participants found in difficult to extrapolate the findings to contemporary interventional practice dominated by stent implantation. This uncertainty stemmed from the lack of clinical trials of abciximab during the stent era. Concerns were also raised that the unrestricted use of two expensive therapeutic modalities (stent implantation and GP IIb/IIIa inhibition) would place severe strains on catheterization laboratory budgets. The general consensus was that, pending the availability of further data, abciximab should probably be reserved for selected at-risk patients. This article summarized the roundtable discussions to provide cardiologists' perspectives on the use of abciximab in interventional practice. An overview of platelet physiology and the rationale for GP IIb/IIIa receptor inhibition; a summary of the results of recent randomized clinical trials that assessed the efficacy of abciximab in PTCA; an account of how stents became the most prevalent technique used in coronary intervention; a summary of the available data evaluating abciximab in conjunction with stent implantation; and a synopsis of the conference discussions are included. PMID- 9738166 TI - Macro-to-micro links in the relation between income inequality and mortality. AB - A growing literature points to links between income inequality and mortality. Any examination of the link should distinguish, both theoretically and empirically, between shifts in inequality that result from changes in the bottom and top of the income distribution. When state-level data from the U.S. censuses of 1980 and 1990 were used to measure differences in mortality, the results indicated that inequality measures reflecting depth of poverty show stronger correlations with mortality than do inequality measures reflecting heights of affluence. In addition, longitudinal data from the Panel Study of Income Dynamics were used to related state-level inequality measures to individual-level data on mortality. This comparison revealed significant associations between degree of income inequality in state of residence and individual risk of death only for nonelderly individuals with middle-class incomes in 1990. PMID- 9738167 TI - Separate but lethal: the effects of economic segregation on mortality in metropolitan America. AB - The increase in income inequality in the United States over the past 20 years has been accompanied by a pronounced increase in economic segregation in urban areas. No research to date has analyzed the potential effects of such spatial segregation on mortality. To investigate these effects, the mortality experience of respondents aged 30 years and older on the 1986-94 National Health Interview Surveys residing in any one of 30 large metropolitan areas in the United States was analyzed. Concentrated poverty was associated with significantly elevated risk of mortality, even after controlling for individual household income. Concentrated affluence showed a consistent, protective effect only among the elderly. The effects were most pronounced among the poor, but were not confined to them. Urban planning should take into account the effects associated with economic residential segregation. PMID- 9738168 TI - Social epidemiology and the fundamental cause concept: on the structuring of effective cancer screens by socioeconomic status. AB - Since the early 1800s, studies have consistently demonstrated that people higher in the socioeconomic hierarchy live longer than people of lower rank. One hypothesis for the persistence of this association is that people who are relatively better off are more able to avoid risks by adopting currently available protective strategies. In a partial test of this idea, the social distributions of two cancer screening tests--Pap smears and mammography--were examined. A review of the literature and an analysis of Behavioral Risk Factor Surveillance System (BRFSS) data showed a consistent association between indicators of socioeconomic status and recent screening. These findings support the theory that societies create and shape patterns of disease. Innovations beneficial to health are carried out within the context of inequalities that shape the distribution of the health benefit, thereby affecting patterns of morality. PMID- 9738169 TI - Contribution of psychosocial factors to socioeconomic differences in health. AB - The National Survey of Mid-life Developments in the United States (MIDUS) is one of several studies that demonstrate socioeconomic gradients in mortality during midlife. When MIDUS findings on self-reported health, waist to hip ratio, and psychological well-being were analyzed for their possible roles in generating socioeconomic differences in health, they revealed clear educational gradients for women and men (i.e., higher education predicted better health). Certain potential mediating variables, like household income, parents' education, smoking behavior, and social relations contributed to an explanation of the socioeconomic gradient. In addition, two census-based measures, combined into an area poverty index, independently predicted ill health. The results suggest that a set of both early and current life circumstances cumulatively contribute toward explaining why people of lower socioeconomic status have worse health and lower psychological well-being. PMID- 9738170 TI - Class, health and justice. AB - Class inequalities in health are intuitively unjust. Although the link between social class and health status has been fully documented, the precise nature of the injustice has not been made clear. Four alternative views are presented, corresponding to four goals: (1) maximizing the sum total of health; (2) equalizing the health status of higher and lower social classes; (3) maximizing the health status of the lowest social class; and (4) maximizing the health status of the sickest individuals in society. The nature of the injustice is further obscured by several theoretical and empirical questions, like the degree and significance of personal responsibility for illness and the relation of the degree of economic inequality to sum total of health. PMID- 9738171 TI - Diffusion of ideas on social inequalities in health: a European perspective. AB - The issue of social inequalities in health has become politically salient during the 1990s in many European countries. Research evidence and ideas that eventually came to the attention of national policy makers helped to trigger this change. The pathways of this information flow are traced through an examination of the work of international agencies and a presentation of case studies from three countries: the Netherlands, Britain, and Sweden. Each country's experience was different but nevertheless influenced and reinforced the courses of action adopted by the others. It is clear that, in order to meet the challenge that health inequalities pose to public health and policy, cooperation will become even more important in the future. PMID- 9738172 TI - Social and economic disparities in health: thoughts about intervention. AB - It is now well established that inequalities in income lead to high morbidity and mortality rates. Certain explanations for this phenomenon are explored: (1) Instead of income inequalities causing disease, the inequalities are determined by powerful cultural forces. (2) The rich get richer and the poor get poorer. (2) Wealthier people can buy the means to protect their health. (4) Poorer people suffer not from poverty, but from relative deprivation. (5) Those with the weakest genes drift into the lower income groups. It is difficult to develop interventions directed to these possibilities. The concept of "control and destiny" is a possible explanation for the relation between inequalities and disease; unlike the other explanations, this idea is amenable to intervention. PMID- 9738173 TI - Palivizumab, a humanized respiratory syncytial virus monoclonal antibody, reduces hospitalization from respiratory syncytial virus infection in high-risk infants. The IMpact-RSV Study Group. AB - OBJECTIVE: To determine the safety and efficacy of prophylaxis with palivizumab in reducing the incidence of hospitalization because of respiratory syncytial virus (RSV) infection in high-risk infants. METHODS: A randomized, double-blind, placebo-controlled trial was conducted at 139 centers in the United States, the United Kingdom, and Canada. During the 1996 to 1997 RSV season, 1502 children with prematurity (less than or equal to 35 weeks) or bronchopulmonary dysplasia (BPD) were randomized to receive 5 injections of either palivizumab (15 mg/kg) or an equivalent volume of placebo by intramuscular injection every 30 days. The primary endpoint was hospitalization with confirmed RSV infection. Children were followed for 150 days (30 days from the last injection). Those with hospitalization as a result of RSV infection were evaluated for total number of days in the hospital, total days with increased supplemental oxygen, total days with moderate or severe lower respiratory tract illness, and incidence and total days of intensive care and mechanical ventilation. The incidence of hospitalization for respiratory illness not caused by RSV and the incidence of otitis media were also evaluated. The placebo and palivizumab groups were balanced at entry for demographics and RSV risk factors. Ninety-nine percent of children in both groups completed the protocol and approximately 93% received all five scheduled injections. RESULTS: Palivizumab prophylaxis resulted in a 55% reduction in hospitalization as a result of RSV (10.6% placebo vs 4.8% palivizumab). Children with prematurity but without BPD had a 78% reduction in RSV hospitalization (8.1% vs 1.8%); children with BPD had a 39% reduction (12.8% vs 7.9%). When gender, entry age, entry weight, BPD, and gestational age were included in a logistic regression model, the effect of prophylaxis with palivizumab remained statistically significant. The palivizumab group had proportionally fewer total RSV hospital days, fewer RSV hospital days with increased oxygen, fewer RSV hospital days with a moderate/severe lower respiratory tract illness, and a lower incidence of intensive care unit admission. Palivizumab was safe and well tolerated. No significant differences were observed in reported adverse events between the two groups. Few children discontinued injections for related adverse events (0.3%). Reactions at the site of injection were uncommon (1.8% placebo vs 2.7% palivizumab); the most frequent reaction was mild and transient erythema. Mild or moderate elevations of aspartate aminotransferase occurred in 1.6% of placebo recipients and 3.6% of palivizumab recipients; for alanine aminotransferase these percentages were 2.0% and 2.3%, respectively. Hepatic and renal adverse events related to the study drug were similar in the two groups. CONCLUSIONS: Monthly intramuscular administration of palivizumab is safe and effective for prevention of serious RSV illness in premature children and those with BPD. PMID- 9738174 TI - Three-year multicenter surveillance of systemic pneumococcal infections in children. AB - OBJECTIVE: To track antibiotic susceptibility of Streptococcus pneumoniae isolates obtained from children with systemic infections and determine outcome of treatment. DESIGN: A 3-year (September 1993 through August 1996) prospective surveillance study of all invasive pneumococcal infections in children. PATIENTS: Infants and children cared for at eight children's hospitals in the United States with culture-proven systemic pneumococcal infection. RESULTS: One thousand two hundred ninety-one episodes of systemic pneumococcal infection were identified in 1255 children. An underlying illness was present in the children for 27% of the episodes. The proportion of isolates that were nonsusceptible to penicillin or ceftriaxone increased annually and nearly doubled throughout the 3-year period; for the last year the percentages of isolates nonsusceptible to penicillin and ceftriaxone were 21% and 9.3%, respectively. There was no difference in mortality between patients with penicillin-susceptible or nonsusceptible isolates. Only 1 of 742 patients with bacteremia had a repeat blood culture that was positive > 1 day after therapy was started. All 24 normal children with bacteremia attributable to isolates resistant to penicillin had resolution of their infection; the most common treatment regimen was a single dose of ceftriaxone followed by an oral antibiotic. CONCLUSIONS: The percentage of pneumococcal isolates nonsusceptible to penicillin and ceftriaxone increased yearly among strains recovered from children with systemic infection. Because empiric antibiotic therapy already has changed for suspected pneumococcal infections, antibiotic resistance has not been associated with increased mortality. Careful monitoring of antibiotic susceptibility and outcome of therapy is necessary to continually reassess current recommendations for treatment. PMID- 9738175 TI - Comparative Efficacy of the Lederle/Takeda acellular pertussis component DTP (DTaP) vaccine and Lederle whole-cell component DTP vaccine in German children after household exposure. Pertussis Vaccine Study Group. AB - BACKGROUND: A household contact substudy was performed as part of a prospective, cohort pertussis vaccine efficacy trial in Germany. DESIGN: Infants received four doses of either the Lederle/Takeda acellular pertussis component diphtheria tetanus toxoids (DTP) vaccine (DTaP) or Lederle whole-cell component DTP vaccine at 3, 4.5, 6, and 15 to 18 months of age (Wyeth-Lederle Vaccines and Pediatrics, Pearl River, NY). An open control group received three doses of diphtheria and tetanus toxoids vaccine (DT) at 3, 4.5, and 15 to 18 months of age. Vaccine efficacy rates were calculated using a number of principal and ancillary case definitions for primary, secondary, and noncases by analyzing secondary attack rates in study infants after exposure to pertussis in the household using 7- to 28- and 7- to 42-day postexposure observation periods and the inclusion and the exclusion of noncases who received macrolide antibiotics or trimethoprim sulfamethoxazole during the exposure period. RESULTS: During a 3.5-year study period, 10271 infants (DTP or DTaP, n = 8532; DT, n = 1739) were enrolled and actively followed along with all household members for cough illnesses. Depending on the case definition, 160 to 519 household exposures to pertussis were identified. In general, secondary attack rates in DT recipients were low and this was primarily because of the frequent use of antimicrobial prophylaxis. Using the principal case definitions and the exclusion of noncases who received macrolide antibiotics or trimethoprim-sulfamethoxazole during the exposure period and the 7 to 42-day observation period, the efficacy of DTP against cough illness of greater than or equal to 7 days duration caused by Bordetella pertussis was 84% (95% confidence interval [CI] = 65-93) and that of DTaP was 58% (95% CI = 30-75). Using similar criteria, the efficacy against typical pertussis (greater than or equal to 21 days of cough with either paroxysms, whoop, or posttussive vomiting) was 94% (95% CI = 77-99) and 86% (95% CI = 62-95) for DTP and DTaP, respectively. The efficacy against any cough illness (with or without) laboratory confirmation was 54% (95% CI = 32-69) and 38% (95% CI = 13-56) for DTP and DTaP, respectively. CONCLUSION: This household contact substudy within our cohort study, with active investigator-generated surveillance, was a severe test of vaccine efficacy. Both vaccines (DTP and DTaP) are better at preventing typical pertussis than mild illness. When case definitions similar to those in other recent trials are used, the Lederle/Takeda vaccine has an efficacy similar to other multicomponent DTaP vaccines. PMID- 9738176 TI - Determinants of health and service use patterns in homeless and low-income housed children. AB - OBJECTIVE: Previous studies of homeless children have described more health problems and service use than in housed children, but failed to control for potential confounding factors that may differ between these children. This observational study examines the relationship of homelessness and other determinants to health status and service use patterns in 627 homeless and low income housed children. METHODS: Case-control study of 293 homeless and 334 low income housed children aged 3 months to 17 years and their mothers conducted in Worcester, Massachusetts. Information was collected about mothers' housing history, income, education, emotional distress, and victimization history. Standardized instruments were administered to assess children's health. Health service use questions were adapted from national surveys. Main outcome measures included health status, acute illness morbidity, emergency department and outpatient medical visits. Multivariable regression analyses were used to examine the association of family and environmental determinants, including homelessness, with health status and service use outcomes. RESULTS: Mothers of homeless children were more likely to report their children as being in fair or poor health compared with their housed counterparts. Homeless children were reported to experience a higher number of acute illness symptoms, including fever, ear infection, diarrhea, and asthma. Emergency department and outpatient medical visits were higher among the homeless group. After controlling for potential explanatory factors, homeless children remained more likely to experience fair or poor health status (adjusted odds ratio [OR] = 2.83; 95% confidence interval [CI], 1.16, 4.87), and a higher frequency of outpatient (OR = 1.71; 95% CI, 1.18, 2.48) and emergency department visits (OR = 1.21; 95% CI, 0.83, 1.74). Mothers' emotional distress was independently associated with acute illness symptoms and frequent use of outpatient and emergency department settings. CONCLUSIONS: Homelessness is an independent predictor of poor health status and high service use among children. The present findings highlight the importance of preventive interventions and efforts to increase access to primary care among homeless children. PMID- 9738177 TI - Children's contributions to pediatric outpatient encounters. AB - OBJECTIVE: Generally, increasing attention is being paid to the quality of doctor patient communication. However, children's contributions have been, until now, primarily ignored in communication research, although there are indications that considering their views increases satisfaction and compliance. In the present study, we examined how children contributed to communication during outpatient pediatric encounters and what factors were associated with children's contributions. PATIENTS: Twenty-one consulting pediatricians videotaped a total of 302 consecutive outpatient encounters. DESIGN: Multilevel analysis was used to take into account the similarity among encounters with the same pediatrician. RESULTS: Children's contributions to the outpatient encounters were limited to 4%. Pediatricians directed one out of every four statements to the child. Although pediatricians asked children a lot of medical questions (26%), only a small part of the medical information (13%) was directed at the children. Apart from social talk and laughter, the amount of pediatrician-child communication increased with children's age. Communication with children suffering from disorders of the nervous system seemed to differ from that with children suffering from other diseases. Allowing children more room in the medical visit did not seem to increase the duration of the visit. CONCLUSIONS: Although recent legislation requires children to be adequately informed, in pediatric outpatient encounters information still tends to be directed primarily at the parents. Children do get the opportunity to talk about social and psychosocial issues. Pediatricians may need to acquire similar communication skills to discuss medical technical issues with the children. PMID- 9738178 TI - Trial on timing of introduction to solids and food type on infant growth. AB - OBJECTIVE: The optimal time and choice of solid foods to introduce to an infant's diet is unknown. The aim of this randomized trial was to determine whether early versus late introduction of solid foods and commercially prepared versus parent's choice of solid foods affects growth or body composition in the first year. METHODS: White infants (n = 165) were recruited before 3 months of age and were randomized to receive: 1) commercially prepared solid foods (commercial) from 3 to 12 months, 2) commercially prepared solid foods from 6 to 12 months, 3) parent's choice of solid foods (choice) from 3 to 12 months, or 4) parent's choice of solid foods from 6 to 12 months. Anthropometrics and body composition, using dual energy x-ray absorptiometry, were determined at 3, 6, and 12 months. Three-day diet diaries were completed at 3, 6, 9, and 12 months. RESULTS: There were no differences in growth or body composition between infants in early versus late introduction groups or commercial versus choice groups at any age. The total energy intake was not different among infants in the early compared with the late group at any age. Infants in the commercial group consumed less protein calories at 9 months (80 +/- 3 kcal/d vs 88 +/- 3 kcal/d) and 12 months 101 +/- 5 kcal/d vs 148 +/- 5 kcal/d), less fat calories at 12 months (263 +/- 10 kcal/d vs 343 +/ 10 kcal/d), and less total calories at 12 months (884 +/- 24 kcal/d vs 1022 +/- 25 kcal/d) compared with the choice group. CONCLUSION: The early introduction of solid foods to an infant's diet does not alter growth or body composition during the first year of life and results in a displacement of energy intake from formula. Infants consuming commercially prepared foods have a decreased caloric intake from protein and fat; however, despite this difference, there is no effect on growth or body composition. PMID- 9738179 TI - Delivery room resuscitation decisions for extremely premature infants. AB - BACKGROUND: Neonatologists are criticized for overtreating extremely premature infants who die despite invasive and costly care. Withholding resuscitation at delivery has been recommended as a way to minimize overtreatment. It is not known how decisions to forgo initiating aggressive care are made, or whether this strategy effectively decreases overtreatment. OBJECTIVE: To identify whether physicians' or parents' preferences primarily determine the amount of treatment provided at delivery, to examine factors associated with the provision of resuscitation, and to assess whether resuscitation at delivery significantly postpones death in nonsurvivors. METHODS: We evaluated delivery room resuscitation decisions and mortality for all infants born at 23 to 26 weeks gestation at the University of North Carolina Hospitals from November 1994 to October 1995. On the day of delivery, the attending neonatologist completed a questionnaire regarding discussion with the parents before delivery, the prognosis for survival estimated before delivery, the degree of certainty about the prognosis, parents' preference for the amount of treatment at delivery, and the degree of influence exerted by parents and physicians on the amount of delivery room treatment provided. Medical records were reviewed for demographics and hospital course. RESULTS: Thirty-one of 41 infants were resuscitated (intubation and/or cardiopulmonary resuscitation) at delivery. Resuscitation correlated with increasing gestational age, higher birth weight, estimated prognosis for survival greater than or equal to 10%, and uncertainty about prognostic accuracy. Physicians saw themselves as primarily responsible for delivery room resuscitation decisions when the parents' wishes about initiating care were unknown, and as equal partners with parents when they agreed on the level of care. When disagreement existed, doctors always thought parents preferred more aggressive resuscitation, and identified parents as responsible for the increased amount of treatment at delivery. Twenty-four infants died before hospital discharge. The median age at death was 2 days when physicians primarily determined the amount of treatment at delivery, 1 day when parents primarily determined the amount of treatment, and < 1 day when responsibility was shared equally. The median age at death was < 1 day when physicians and parents agreed about the preferred amount of treatment at delivery and 1.5 days when they disagreed. The median age at death was < 1 day when parents' preferences were known before delivery and 4 days when parents' preferences were unknown. CONCLUSIONS: Physicians resuscitated extremely premature infants at delivery when they were very uncertain about an infant's prognosis or when the parents' desires about treatment were unknown. When parents' preferences were known, parents usually determined the amount of treatment provided at delivery. Resuscitation at delivery usually postponed death by only a few days, decreasing prognostic uncertainty and honoring what physicians perceived were parents' wishes for care, without substantially contributing to overtreatment. PMID- 9738180 TI - Heterogeneity of diagnoses presenting as cyclic vomiting. AB - OBJECTIVE: To establish the diagnostic profile in children who present with cyclic vomiting. METHODS: We studied 225 children < 18 years of age who presented to our pediatric gastroenterology service from 1986 to 1997 with at least three discrete episodes of vomiting between which they were well. To determine the diagnoses in those presenting with a pattern of cyclic vomiting, the results of diagnostic testing and responses to various treatments were obtained from a combination of chart review and structured telephone interviews. RESULTS: The largest diagnostic category included idiopathic cyclic vomiting syndrome (88%). Extraintestinal disorders (7%) and gastrointestinal disorders (5%) constituting the probable cause of vomiting were established in those having complete cessation of episodes after therapy. In 12%, serious surgical disorders of the gastrointestinal (malrotation), renal (acute hydronephrosis), and central nervous systems (neoplasm) were found. In 2%, serious endocrine (Addison's disease) and metabolic disorders (disorder of fatty acid oxidation) were found. Among those with idiopathic cyclic vomiting syndrome, 41% had associated disorders (gastroesophageal reflux and chronic sinusitis) that could contribute to the vomiting, but, based on a partial response to therapy, were not deemed to be the main cause. Altogether 49% had an identified disorder that probably caused or could contribute to the vomiting. CONCLUSIONS: The cyclic pattern of vomiting is a symptom complex that can be induced by heterogeneous disorders that either cause or contribute to the vomiting. Once the cyclic vomiting pattern is identified, systematic diagnostic testing is warranted to look for these underlying disorders. PMID- 9738181 TI - Adolescent medicine training in pediatric residency programs: are we doing a good job? AB - OBJECTIVES: To determine how pediatric residency programs are responding to the new challenges of teaching adolescent medicine (AM) to residents by assessing whether manpower is adequate for training, whether AM curricula and skills are adequately covered by training programs, what types of teaching methodologies are used to train residents in AM, and the needs for new curricular materials to teach AM. DESIGN: A 3-part 92-item survey mailed to all US pediatric residency training programs. SETTING: Pediatric residency programs. PARTICIPANTS: Residency program directors and directors of AM training. MAIN OUTCOME MEASURES: AM divisional structure, clinical sites of training, presence of a block rotation, and faculty of pediatric training programs; training materials used and desired in AM; perceived adequacy of coverage of various AM topics; competency of residents in performing pelvic examinations in sexually active teens; and manpower needs. RESULTS: A total of 155/211 (73.5%) of programs completed the program director and the AM parts of the survey. Ninety-six percent of programs (size range, 5-120 residents) had an AM block rotation and 90% required the AM block; those without a block rotation were more likely to be larger programs. Only 39% of programs felt that the number of AM faculty was adequate for teaching residents. Almost half of the programs reported lack of time, faculty, and curricula to teach content in substance abuse. Besides physicians, AM teachers included nurse practitioners (28%), psychologists (25%), and social workers (19%). Topics most often cited as adequately covered included sexually transmitted diseases (81.9%), confidentiality (79.4%), puberty (77.0%), contraception (76.1%), and menstrual problems (73.5%). Topics least often cited as adequately covered included psychological testing (16.1%), violence in relationships (20.0%), violence and weapon-carrying (29.7%), and sports medicine (29.7%). Fifty-eight percent of 137 respondents thought that all or nearly all of their residents were competent in performing pelvic examinations by the end of training; there was no difference between perceived competence and the residents' use of procedure books. Seventy-four percent used a specific curriculum for teaching AM; materials included chapters/articles (85%), lecture outlines (76.1%), slides (41.9%), videos (35.5%), written case studies (24.5%), computerized cases (6.5%), and CD-ROMs (3.2%). Fifty-two percent used Bright Futures, 48% used the Guidelines for Adolescent Preventive Services, and 14% used the Guide to Clinical Preventive Services for teaching clinical preventive services. Programs that used Bright Futures were more likely to feel that preventive services were adequately covered in their programs than those who did not (78% vs 57%). A majority of programs desired more learner-centered materials. CONCLUSIONS: Although almost all pediatric programs are now providing AM rotations, there is significant variability in adequacy of training across multiple topics important for resident education. Programs desire more learner centered materials and more faculty to provide comprehensive resident education in AM. PMID- 9738182 TI - Hidden spears: handlebars as injury hazards to children. AB - OBJECTIVES: To delineate the mechanism of serious bicycle handlebar-related injuries in children and make recommendations for preventive strategies. METHODS: Prospective cross-sectional surveillance system of seriously injured child bicyclists supplemented by in-depth, on-site crash investigation to delineate specific injury mechanisms. Interdisciplinary analyses involved engineers, clinicians, epidemiologists, and biostatisticians. SETTING: The emergency department and in-patient trauma service of an urban level one pediatric trauma center between October 1995 and September 1997. PARTICIPANTS: Patients under 18 years of age who were treated for serious bicycle-related injuries (Abbreviated Injury Scale scores of 2 or greater). RESULTS: The surveillance system identified two distinct circumstances for serious child bicyclist injury: 1) handlebar related injuries associated with minor incidents (falls from bicycles) and 2) nonhandlebar-related injuries associated with severe incidents (bicycle-motor vehicle crashes). Crash investigations explored the minor incidents that resulted in serious handlebar-associated injuries. In the typical mechanism, as the child lost control of the bicycle and began to fall, the front wheel rotated into a plane perpendicular to the child's body. The child then landed on the end of the handlebar resulting in serious truncal injuries. CONCLUSIONS: A discordancy exists between the apparently minor circumstances and serious injuries sustained by child bicyclists who impact bicycle handlebars. Recognition of the mechanism of handlebar-related injuries might aid the practitioner in early diagnosis of serious abdominal injuries in child bicyclists. This injury mechanism may be avoided through bicycle redesign that would involve both limiting rotation of the front wheel and modifying the ends of handlebars. An integrated approach involving a surveillance system to identify an injury hazard supplemented by in depth, on-site crash investigations effectively provided the detailed mechanism of injury needed to develop interventions. PMID- 9738183 TI - Height as a marker of childhood development and late-life cognitive function: the Honolulu-Asia Aging Study. AB - OBJECTIVE: Growing evidence suggests that structural and functional brain reserves, thought to develop in childhood and adolescence, may be crucial in determining when cognitive impairment begins. The purpose of this report is to examine the relationship of height, as a marker of childhood development, to late life cognitive function in a sample of elderly Japanese-American men. METHOD: Cognitive performance was assessed from 1991 to 1993 in the Honolulu-Asia Aging Study in 3733 men aged 71 to 93 years and related to height that was measured 25 years earlier. RESULTS: Among the study sample, shorter men were older, leaner, and less educated than taller men. Shorter men also spent more years of their childhood living in Japan and were more likely to have had fathers in unskilled professions. After adjustment for age, the prevalence of poor cognitive performance declined consistently with increasing height from 25% in men shorter than 154 cm (61 in) to 9% in those taller than 174 cm (69 in). Excluding men with stroke or dementia did not alter the association between height and cognitive performance. Apolipoprotein E4 was unrelated to height and did not effect the association between height and cognitive function. The prevalence of Alzheimer's disease was higher in men who were 154 cm (61 in) or shorter as compared with men who were taller (4.7% vs 2.9%, respectively). There was no association between height and vascular dementia. CONCLUSION: Efforts to improve prenatal and early life conditions to maximize growth in childhood and adolescence could diminish or delay the expression of cognitive impairments that occur later in life. Prevention of some late-life cognitive impairments may have pediatric origins. PMID- 9738184 TI - High-expenditure children with Supplemental Security Income. AB - OBJECTIVE: To examine the clinical characteristics and health service use of children with high Medicaid expenditures. METHODOLOGY: We examined 1992 Medicaid claims and eligibility files from four states (California, Georgia, Michigan, Tennessee) for children with at least $10000 billed to Medicaid who obtained Medicaid through the Supplemental Security Income (SSI) Program and a comparison group (matched by age group and gender) of children receiving Medicaid for other reasons. We compared mean expenditures, examined expenses by category, and examined diagnoses associated with at least $10000 in expenses. RESULTS: In 1992, Medicaid paid on average approximately $1000 for children with non-SSI Medicaid enrollment. Expenditures for children with SSI were 2.9 to 9.4 times higher, but once the approximately 10% of children with high expenditures were excluded, SSI average expenditures were only 1.5 to 2.7 times higher than the non-SSI average. Children with high expenditures are likely to use hospitals and long-term care, and these services account for more than half of the average expenditures. Children with high expenditures and SSI are more likely to have chronic medical conditions than are their peers enrolled in Medicaid but not through SSI. CONCLUSIONS: A small proportion of children, even on SSI, account for very large proportions of Medicaid expenditures. Most children with SSI, despite having relatively severe mental health, physical, or developmental disabilities, have relatively modest Medicaid expenditures. PMID- 9738185 TI - Sleep-disordered breathing and school performance in children. AB - OBJECTIVE: To assess the impact of sleep-associated gas exchange abnormalities (SAGEA) on school academic performance in children. DESIGN: Prospective study. SETTING: Urban public elementary schools. PARTICIPANTS: Two hundred ninety-seven first-grade children whose school performance was in the lowest 10th percentile of their class ranking. METHODS: Children were screened for obstructive sleep apnea syndrome at home using a detailed parental questionnaire and a single night recording of pulse oximetry and transcutaneous partial pressure of carbon dioxide. If SAGEA was diagnosed, parents were encouraged to seek medical intervention for SAGEA. School grades of all participating children for the school year preceding and after the overnight study were obtained. RESULTS: SAGEA was identified in 54 children (18.1%). Of these, 24 underwent surgical tonsillectomy and adenoidectomy (TR), whereas in the remaining 30 children, parents elected not to seek any therapeutic intervention (NT). Overall mean grades during the second grade increased from 2.43 +/- 0.17 (SEM) to 2.87 +/- 0.19 in TR, although no significant changes occurred in NT (2.44 +/- 0.13 to 2.46 +/- 0.15). Similarly, no academic improvements occurred in children without SAGEA. CONCLUSIONS: SAGEA is frequently present in poorly performing first-grade students in whom it adversely affects learning performance. The data suggest that a subset of children with behavioral and learning disabilities could have SAGEA and may benefit from prospective medical evaluation and treatment. PMID- 9738186 TI - Richmond Award acceptance speech. PMID- 9738187 TI - Training pediatricians to become child advocates. PMID- 9738188 TI - A subspecialist's view of training and pediatric practice in the next millennium. PMID- 9738190 TI - Intravenous immune globulin shortage: experience at a large children's hospital. PMID- 9738189 TI - Decision-making in delivery room resuscitation: a team sport. PMID- 9738191 TI - Human hepatocyte transplantation: gene therapy and more? PMID- 9738192 TI - Humanized monoclonal antibody for prevention of respiratory syncytial virus infection. PMID- 9738193 TI - Unrecognized microscopic hyphema masquerading as a closed head injury. AB - OBJECTIVE: To present a child with an unrecognized microscopic traumatic hyphema and acute glaucoma who was initially treated as a closed head injury patient. DESIGN: Case report and discussion. RESULTS: Symptoms attributable to unrecognized occult ocular injury in a child with sickle cell trait resulted in evaluation and treatment of the child for a closed head injury. Evaluation included a computed tomography scan of the head and lumbar puncture. An ophthalmologic consultation later revealed a microscopic hyphema and acute glaucoma as the etiology of the child's signs and symptoms. CONCLUSIONS: Children who present with neurologic symptoms and a history of ocular trauma should undergo an ophthalmologic examination as soon as possible. Hyphema, even if not readily visible on physical examination, can result in the development of acute glaucoma with signs and symptoms that resemble a closed head injury. PMID- 9738194 TI - Colovesical fistula resulting from a perforated colonic duplication. AB - Colovesical fistulas in children are most often associated with high anorectal imperforations. Acquired enterovesical fistulas in children only rarely have been reported as a consequence of an inflammatory process. We present a case of an acquired colovesical fistula formed by the erosion of an abscess at the distal end of a colonic duplication in a child who presented with fever of unknown origin. PMID- 9738195 TI - Managed care and children with special health care needs: a subject review. American Academy of Pediatrics Committee on Children with Disabilities. AB - Barriers to access to health care frequently overshadow the opportunities for improvement through managed care, especially regarding children with special health care needs. This statement discusses such opportunities, identifies challenges, and proposes active roles for pediatricians, and families of patients to improve some aspects of managed care for children with special health care needs. PMID- 9738196 TI - Questions concerning cough preparation statement by the AAP Committee on Drugs. PMID- 9738197 TI - Blowing bubbles, calming patients. PMID- 9738198 TI - Interventions for perinatal hypoxic-ischemic encephalopathy. PMID- 9738199 TI - Bedsharing promotes breastfeeding and AAP Task Force on Infant Positioning and SIDS. PMID- 9738200 TI - Newborn hearing screening and hypothyroidism. PMID- 9738201 TI - What is the role of environmental health science in cancer prevention? PMID- 9738202 TI - Value of monitoring serum immunoglobulins questioned. PMID- 9738203 TI - PAH cleanup in Illinois. PMID- 9738204 TI - Prevention of environmentally related diseases. PMID- 9738206 TI - Science and judgement in risk assessment: needs and opportunities. PMID- 9738205 TI - TCDD carcinogenicity in humans. PMID- 9738207 TI - Estimating the effects of toxicants on ecosystem services. AB - Numerous functions of ecosystems are essential to the quality of human life, including the provision of food, the decomposition of sewage, the provision of portable water, and the replacement of breathable air. Although attributes of ecosystems directly of use to human societies are not the only ones worth protecting, emphasizing their services may be the most effective means of communicating risks of toxicants to the general public. However, although spatial and temporal scales of experiments to assess risk vary relatively little, actual spatial scales vary considerably, from local environments to global ecosystems. Generally, models are used to bridge these gaps in scale. In this paper, we examine ways in which toxicity test endpoints have been developed to describe effects of pollutants on essential ecosystem functions and the ways in which results are then extrapolated to scales that risk managers can use. PMID- 9738209 TI - Three-dimensional visualization of physiologically based kinetic model outputs. AB - Outputs from a physiologically based toxicokinetic (PB-TK) model for fish were visualized by mapping time-series data for specific tissues onto a three dimensional representation of a rainbow trout. The trout representation was generated in stepwise fashion: 1) cross-section images were obtained from an anesthetized fish using a magnetic resonance imaging system, 2) images were processed to classify tissue types and eliminate unnecessary detail. 3) processed images were imported to a visualization software package (Application Visualization System) to create a three-dimensional representation of the fish, encapsulating five volumes corresponding to the liver, kidney, muscle, gastrointestinal tract, and fat, Kinetic data for the disposition of pentachloroethane in trout were generated using a PB-TK model. Model outputs were mapped onto corresponding tissues volumes, representing chemical concentration as color intensity. The workstation software was then used to animate the images, illustration the accumulation of pentachloroethane in each tissue during a continuous branchial (gill) exposure. PMID- 9738208 TI - The biological speciation and toxicokinetics of aluminum. AB - This review discusses recent literature on the chemical and physiological factors that influence the absorption, distribution, and excretion of aluminum in mammals, with particular regard to gastrointestinal absorption and speciation in plasma. Humans encounter aluminum, a ubiquitous yet highly insoluble element in most forms, in foods, drinking water, and pharmaceuticals. Exposure also occurs by inhalation of dust and aerosols, particularly in occupational settings. Absorption from the gut depends largely on pH and the presence of complexing ligands, particularly carboxylic acids, with which the metal can form absorbable neutral aluminum species. Uremic animals and humans experience higher than normal body burdens of aluminum despite increased urinary clearance of the metal. In plasma, 80-90% of aluminum binds to transferrin, an iron-transport protein for which receptors exist in many tissue. The remaining fraction of plasma aluminum takes the form of small-molecule hydroxy species and small complexes with carboxylic acids, phosphate, and, to a much lesser degree, amino acids. Most of these species have not been observed in vivo but are predicted from equilibrium models derived from potentiometric methods and NMR investigations. These models predict that the major small-molecule aluminum species under plasma conditions are charged and hence unavailable for uptake into tissues. PMID- 9738210 TI - Gastrointestinal upsets associated with ingestion of copper-contaminated water. AB - During 1992 and 1993 the Wisconsin Division of Health investigated five cases in which copper-contaminated drinking water was suspected of causing gastrointestinal upsets. Each of these case studies was conducted after our office was notified of high copper levels in drinking water or notified of unexplained illnesses. Our findings suggest that drinking water that contains copper at levels above the federal action limit of 1.3 mg/l may be a relatively common cause of diarrhea, abdominal cramps, and nausea. These symptoms occurred most frequently in infants and young children and among resident of newly constructed or renovated homes. PMID- 9738211 TI - Congener-specific levels of dioxins and dibenzofurans in U.S. food and estimated daily dioxin toxic equivalent intake. AB - Food, especially meat, milk, and fish, is the immediate source of almost all polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs), and dioxinlike compounds in the general population. To estimate intake of these highly toxic compounds, we performed congener-specific dioxin analyses for the first time on U.S. food for 18 dairy meat, and fish samples from a supermarket in upstate New York. 2,3,7,8 Tetrachlorodibenzo-p-dioxin (TCDD, "dioxin") toxic equivalents (TEqs) on a wet weight basis for the dairy products ranged for 0.04 to 0.7 ppt, meat TEqs ranged from 0.03 to 1.5 ppt, and fish TEqs ranged from 0.02 to 0.13 ppt. Previous human breast milk and infant formula analyses were used with the current preliminary food data to estimate a range of dioxin intake for Americans. Average daily food intake of TEqs for an adult weighing 65 kg was estimated to be between 0.3 and 3.0 pg/kg body weight, for a total of 18-192 pg TEq, using 1986 American consumption rates. Due to the relatively high level of PCDDs and PCDFs commonly found in human breast milk from American women and from women in other industrial countries, a nursing infant may consume an average of 35-53 pg TEq/kg body weight/day in its first year of life. This may be compared with the current U.S. EPA virtually safe dose of 0.006 pg TCDD/kg body weight per day over a 70 year lifetime based on an upper limit cancer risk of 10(-6), or the 10 pg/kg/day used by some European government agencies. PMID- 9738212 TI - Influence of short-term dietary measures on dioxin concentrations in human milk. AB - Breast-feeding may expose infants to high levels of toxic chlorinated dioxins. To diminish intake of these lipophilic compounds by the baby, two diets were tested for their ability to reduce concentrations of dioxins in human milk. The diets were a low-fat/high- carbohydrate/low-dioxin diet. (about 20% of energy intake derived from fat) and a high fat /low-carbohydrate/low-dioxin diet. These diets were tested in 16 and 18 breast-feeding women, respectively. The test diets were followed for 5 consecutive days in the fourth week after delivery. Milk was sampled before and at the end of the dietary regimen, and dioxin concentrations and fatty acid concentrations were determined. Despite significant influences of these diets on the fatty acid profiles, no significant influence on the dioxin concentrations in breast milk could be found. We conclude that short-term dietary measures will not reduce dioxin concentration in human milk. PMID- 9738213 TI - Comparison of the toxic effects of hydrogen peroxide and ozone on cultured human bronchial epithelial cells. AB - In this study, we compared the cytotoxic and genotoxic effects of hydrogen peroxide and ozone on cultured human airway epithelial cells in primary culture. Both agents caused a dose-dependent loss in the replicative ability of epithelial cells and at higher levels of exposure caused acute cytotoxicity as measured by release of lactate dehydrogenase. Differences were seen, however, between the agents' effects with regard to induction of DNA single strand breaks as measured by alkaline elution:; whereas single-strand breaks were detected in significant amounts at concentration of hydrogen peroxide that cause acute cytotoxicity, none were detected at any of the levels of ozone exposure examined. A difference was also seen in the ability of the iron chelator deferoxamine to protect cells from the effect of the two oxidants. Preincubation of cultures with deferoxamine appreciably attenuated the toxicity of hydrogen peroxide but not of ozone. These data suggest that ozone has significant toxic effects on bronchial epithelial cells not mediated through the generation of hydrogen peroxide or hydroxyl radical. Furthermore, the data indicate that the inhibiting action of ozone on cell replicative ability is not mediated through a mechanism related to DNA single strand breaks. PMID- 9738214 TI - [Sentinel lymphadenectomy in malignant melanoma]. AB - The sentinel lymph node dissection (SLND) is one of the most striking developments in the treatment of melanoma. Since the first report by Morton et al. in 1992, the method has been refined, and its use has increased. Introduced as an alternative to elective lymph node dissection (ELND), it has rapidly made its way into clinical practice. SLND allows precise pathologic staging through removal and analysis of a limited number of nodes (false-negative rate < 2%). It distinguishes patients with clinically occult nodal disease from those with tumor free regional basin who would not benefit from radical dissection. However, the SLND is still an experimental procedure with yet unproven utility. PMID- 9738215 TI - [Sentinel node detection in breast carcinoma]. AB - The aim of sentinel node biopsy (SN) in breast cancer patients is to detect the tumor-draining lymph node by means of isosulfan blue or 99mTc-labelled colloids. SN can be detected in 80 to 85% of the patients, depending on the size of the tumor. Preoperative lymphoscintigraphy permits the draining nodes along the internal mammary artery also to be visualized. The predictive value of the histological findings of SN for lymph nodes obtained from axillary dissection is about 95%. Because of different diagnostic procedures using various radiotracers each center has to follow its own learning curve. To be sure that the nodal status derived from a SN procedure is of identical value to axillary dissection about 100 patients have to undergo sentinel node detection, followed by axillary dissection, and concordant results should be obtained in 95% of them at least. The SN, however, offers a chance of assessing the lymph node at risk for metastasis by more detailed histological procedures. Morbidity as a result of treatment for primary breast cancer can be decreased if only patients suffering from metastatic disease are subjected to axillary dissection. Currently, the indication criteria for a SN procedure should be restricted to small tumors (T1) with clinically uninvolved axillary status and patients with ductal carcinoma in situ (DCIS). PMID- 9738216 TI - [Lymphadenectomy in colorectal carcinoma]. AB - Recurrence of colorectal carcinomas occurs in about 50% of the cases with localized neoplasia. It is understood that the tumor recurrence is due to residual micrometastases not found during surgery or extraregional (peripheral blood or bone marrow). We developed a procedure to detect non-visible, abdominal metastases using a radiolabeled anti-tumor cell antibody injected before the operation (radioimmunoguided surgery RIGS). However, even with the best technique, it is not possible to remove all micrometastasis if a hematogenic dissemination happens. Based on the knowledge of disturbing humoral immune reaction is mounted against shed tumor associated antigens (TAA), we developed a new method to reduce and correct the B cell response and B cell recruitment due to chronic TAA immun complex presentation on follicular dendritic cells (immune corrective surgery, ICS). This method is based on a selective lymphadenectomy. The target lymph nodes were those loaded with TAA-immune complex. The detection method used was the injection of radiolabeled antibody able to recognize the immune complex. From 20 patients (stage I, II and III) treated with ICS, 17 survived more than 5 years 'showing a statistically significant increase of survival compared to patients treated with standard procedures. In conclusion, these data show that surgery of colorectal cancer should be selectively extended to specific anatomical regions in order to remove hidden micrometastases, and more importantly, correct postoperative immune processes that could suppress the T cell response against residual tumor cells. PMID- 9738217 TI - [Is the stapled suture in visceral surgery still justified? A prospective controlled, randomized study of cost effectiveness of manual and stapler suture]. AB - Hospitals are facing increasing economic pressure. It therefore seems necessary to evaluate the efficiency and effectiveness of medical or surgical interventions. In this study 324 anastomoses (167 stapled and 157 hand-sewn) were performed after randomization during 200 elective operations [20.5% gastrectomies, 14% gastric resections (Billroth II), 15% Whipple's procedures, 4% segmental colonic resections, 18% right-sided hemicolectomies, 4% left-sided hemicolectomies, 22% sigmoid- or anterior rectal resections, 2.5% total colectomies with pouch-anal anastomoses] in 200 patients. Postoperative motility (time to full oral diet, time with naso-gastric tube) and hospitalization were comparable in both groups. Anastomotic insufficiency was observed in 2.1% of all patients, five after stapled and two after hand-sewn anastomoses. Hospital mortality was 1.5%. All stapled anastomoses were performed significantly (P < 0.001) faster. However, the cost of material for these anastomoses was significantly (P < 0.001) higher, resulting in significantly higher total costs for reconstruction. The time saving for the reconstruction did not influence the total operative time (except for stapled gastrectomy). Therefore, all operations with stapled reconstruction were more expensive than those with sutured reconstruction. The difference was significant for the gastrectomy (P < 0.01), colonic resection (P < 0.01) and sigmoid and rectal resection (P < 0.001) groups. Stapled and sutured anastomoses are equally effective. Stapled anastomoses are not efficient, however, and should be reserved for individual indications. PMID- 9738218 TI - [Significant extension of survival by complete resection of isolated lung metastases after breast carcinoma]. AB - At least 25% of breast cancer patients develop distant metastases. In spite of increasingly sophisticated palliative therapies, the survival time of patients with metastasis did not appear to be significantly prolonged during the last 25 years (19-32 months following diagnosis) and 95% of them die from metastatic disease. Therefore, it seems appropriate that the therapeutic risk/benefit ratio and impact on quality of life should be reassessed when asymptomatic patients are treated. Surgical treatment and pulmonary resection for metastatic disease has been proven a valuable therapeutic concept for a variety of malignancies. Three epidemiologically comparable collectives out of a total of 125 patients from our clinic were treated for isolated pulmonary metastasis following breast cancer (observation period: 1977-1997). Complete data sets could be established for 96 patients and were retrospectively analyzed following stratification into three groups according to their surgical therapy. Twenty-eight patients underwent complete resection (K), 34 had incomplete resections (I) and 34 had no surgical intervention for lung metastases (N). Comparison of the three therapy arms concerning stage, histology and receptor levels of the primary tumor, number of metastases, and the disease-free interval yielded no significant differences between groups K, I and N. Patients after complete resection of isolated lung metastases (group K) had a mean survival of 79 months (5-year survival 80%, 10 year survival 60%). This was significantly better than groups I and N (P < 0.00002). The mean survival of groups I and N was not significantly different (15.5 and 9 months respectively). The disease-free interval after operation of the primary tumor had no impact on the survival of group K, but showed a high correlation with the survival of group N (R2 = 0.81). Complete resection of isolated pulmonary metastases from carcinoma of the breast results in marked prolongation of survival with a low morbidity rate. Hence, routine chest X-ray should be considered an indispensable part of the oncological aftercare in breast cancer patients. PMID- 9738219 TI - [Adjuvant regional arterial port chemotherapy after resection of colorectal liver metastases]. AB - Between 1986 and 1995 we performed radical hepatic resections (R0 resections) in 109 patients with hepatic metastases following colorectal carcinoma. In 50 patients a hepatic arterial port device was implanted for adjuvant regional chemotherapy (HAI). Mitomycin C, 5-fluorouracil, and since 1993 folinic acid have been administered during 6 monthly repeated courses. In 9 patients, the treatment had to be withdrawn because of complications. The remaining 59 patients were not treated. In 73% of the patients after port implantation mostly minor complications occurred during chemotherapy. Our results confirmed a markedly increased survival rate during the first 3 postoperative years, followed by a prolongation of median survival time of treated patients compared to untreated patients. Nevertheless, the observed differences of median survival were not statistically different. In contrast, the 5-year survival rates of both groups were not different. The frequency, localization, and resectability of recurrences were not influenced by adjuvant chemotherapy. However, the lengthening of mean survival time in the treated group might reflect a delay in the occurrence of early recurrences. In conclusion, adjuvant hepatic arterial chemotherapy following resection of colorectal hepatic metastases might be able to prolong the time until recurrence, but does not help to avoiding it. Therefore, it did not increase the rate of cure following R0 resections of colorectal hepatic metastases in our series. Taking into account the high rate of local complications of the port systems in our series, angiographic controls are strongly recommended prior to each chemotherapeutic cycle. PMID- 9738221 TI - [Coarctation of the thoracoabdominal aorta]. AB - Coarctation of the thoracic or abdominal aorta often is associated with reduced perfusion of one or both kidneys, resulting in severe renovascular hypertension, which significantly influences the spontaneous course of these patients. Fourteen of 15 patients who were operated upon between 1983 and 1996 suffered from arterial hypertension. Thirteen patients had ischemia of one or both kidneys resulting from renal artery stenosis or stenosis of the descending or visceral aorta. Because of a hemodynamically significant stenosis the aorta was reconstructed in 11 patients by interposition graft, bypass, or patchplasty. Concerned renal or visceral arteries were reconstructed by bypass/interposition graft, patchplasty, or reimplantation. One patient died, presumably from septic bleeding, 3 weeks post operatively. There was a cure of hypertension in 3 and an improvement in 6 patients. In the individual patient hypertension could be cured more often if the diagnosis of coarctation were established early. PMID- 9738220 TI - [Changes in the clinical picture and surgical therapy of Crohn disease. 10 years experiences]. AB - Changes in clinical presentation of Crohn's disease (CD), patients' age at the time of disease manifestation and operation, general condition and duration of medical management were investigated in a retrospective study. 395 operations for CD were divided into consecutive groups and analysed. In an increasing number of cases patients presented with the "fibrostenotic type" rather than the "fistulizing type" of complaints. The duration of disease and of medical management increased. Patients in later groups were younger and in better general condition. We found a rise in frequency of patients with recurrent CD or prior abdominal surgery for other reasons. Primary surgery for CD increasingly consisted of resection of solitary segments of gut; in the case of recurrence, of multiple, minimal resections. The average total length of the resected intestine could be significantly reduced. The complication rate was equal in elective and emergency surgery. PMID- 9738222 TI - [This can't go on--ambulatory surgeons protest]. PMID- 9738223 TI - [Treatment exceeding the limit of mandated health insurance]. PMID- 9738224 TI - [RKI "Current version of prophylactic drug therapy after occupational HIV exposure". Consensus conference 21 November 1997 in Munich]. PMID- 9738225 TI - [Brief preoperative radiotherapy in surgical therapy of rectal carcinoma. Long term results of a prospective randomized study]. AB - To ascertain whether preoperative short-term radiotherapy can improve local tumor control and the long-term survival of patients with operable rectal cancer, a prospective randomised trial was performed from 1988 to 1993. Ninety-three patients with rectal cancer were either directly treated with surgery (n = 46) or underwent preoperative radiotherapy with 5 x 3.3 Gy irradiation and operation within 48 h (n = 47). If indicated (T4, UICC stage III) patients also received postoperative irradiation. Comparison of the methods of operation (abdominoperineal amputation versus anterior resection) revealed no significant difference in 5-year survival rate (P = 0.393). Local control of R0-resected tumors was improved after preoperative irradiation (P = 0.08). The 5-year survival rate was significantly higher after preoperative short-term radiotherapy (P = 0.027). Preoperative radiotherapy is not an independent factor according to overall survival (P = 0.078) and local recurrence (P = 0.07). In agreement with the results of other authors the present study indicates improved local tumor control of rectal cancer after preoperative radiation therapy. The 5-year survival rate was significantly better after preoperative radiotherapy than after surgery alone. PMID- 9738226 TI - [Management of complicated incisional hernias with underlay-technique implanted polypropylene mesh. An effective technique in French hernia surgery]. AB - Incisional hernia repair with conventional techniques (simple closure, Mayo technique) is associated with unacceptable recurrence rates of 30-50%. Therefore, surgical repair using different prosthetic biomaterials is becoming increasingly popular. Further to favourable results by French hernia surgeons, we studied the results of underlay prosthetic mesh repair using polypropylene mesh in complicated and recurrent incisional hernias. METHOD: After preparation and excision of the entire hernia sac, the posterior rectus sheath is freed from the muscle bellies on both sides. The peritoneum and posterior rectus sheaths are closed with a continuous looped polyglyconate suture. The prosthesis used for midline hernias is positioned on the posterior rectus sheath and extends far beyond the borders of the myoaponeurotic defect. The anterior rectus sheath is closed with a continuous suture. The prosthesis for lumbar and subcostal hernias is placed in a prepared space between the transverse and oblique muscles. Intraperitoneal placement of the mesh must be avoided. RESULTS: Between January 1996 and August 1997 we performed a total of 33 incisional hernia repairs (14 primary hernias, 19 recurrent hernias) using this technique (16 women, 17 men, mean age 56.19 +/- 12.92 years). Local complications occurred in four patients (12%): superficial wound infection (n = 2), postoperative bleeding, requiring reoperation (n = 1), minor hemato-seroma (n = 1). One patient suddenly died on the 3rd post-operative day from severe pulmonary embolism (mortality 3%). Twenty two patients with a minimum follow up to 6 months were re-examined clinically. The average follow-up time for this group was 9 months (range 6-17 months). To date no recurrent hernias have been observed. There were only minor complaints like "a feeling of tension" in the abdominal wall (n = 3) and slight pain under physical stress (n = 6). CONCLUSIONS: The use of prosthetic mesh should be considered for repair of large or recurrent incisional hernias, especially in high-risk patients (obesity, obstructive lung disease) and complicated hernias. The aforementioned technique of underlay prosthetic repair using polypropylene mesh fixed onto the posterior rectus sheath allows for anatomical and consolidated reconstruction of the damaged abdominal wall with excellent results and low complication rates. PMID- 9738227 TI - [Fatal carbon dioxide embolism as a complication of endoscopic interventions]. AB - Fatal complications during laparoscopy mostly originate from injury to major pelvic vessels, causing severe hemorrhage or carbon dioxide embolism. We report a case of a 30-year-old patient who--after unsuccessful resuscitation--died during gynecologic laparoscopy. The noted signs corresponded to acute gas embolism, particularly as ultrasound revealed intravascular gas pulsation. At autopsy, 2 days post mortem, a puncture of the left common iliac vein was discovered. Despite this, gas bubbles in the right heart could not be confirmed. Pre conditions for fatal carbon dioxide embolism are stressed. To prove the presence of carbon dioxide gas post mortem, autopsy has to be performed as soon as possible and the corpse has to be stored without cooling. PMID- 9738228 TI - [Yamakawa prosthesis with prolonged placement as a therapy concept exemplified by benign biliary stricture]. AB - AIM: As an alternative method to the operative revision of benign bile duct strictures, we report the use of a large-bore bile duct prosthesis (Yamakawa prosthesis) in one patient. METHODS: Bilateral percutaneous transhepatic implantation of Yamakawa prostheses (14 F right side, 12 F left side) was performed without adjunctive balloon dilatation. The prostheses were exchanged every 8 weeks under continuous antibiotic treatment and were finally removed after 8 months. RESULTS: Control cholangiography showed a normal intrahepatic biliary system on the right side and a 50% stenosis at the orifice of the left hepatic duct. Follow-up over 18 months showed no signs of recurrent disease. CONCLUSIONS: In comparison to balloon dilatation and implantation of metallic stents, prolonged bilateral splinting with large-bore Yamakawa prostheses seems to be an alternative for the treatment of benign bile duct strictures. PMID- 9738229 TI - [Cavernous hemangioma of the rectum--a rare cause of peranal hemorrhage]. AB - We report the case of a 34-year-old woman with severe rectal bleeding since the age of 17. The cause of the bleeding was a cavernous haemangioma of the rectum. The extent of the disease was not realised for many years. Sclerosing injections, laser coagulation and even suture ligation were helpful in acute bleeding episodes but did not result in definitive healing. Finally cure was achieved by resection of the rectum and colo-anal sleeve anastomosis. The clinical presentation and the management are described and discussed. PMID- 9738230 TI - [Acute ischemia of the extremities]. PMID- 9738231 TI - [The Nobel Prize for chemistry in 1997--mitochondrial ATP synthase]. PMID- 9738232 TI - [Nobel Prize for Jens Christian Skou for the discovery of Na(+)-K+ ATPase]. PMID- 9738233 TI - [Molecular mechanisms of apoptosis induced by activation of membrane receptors from the TNF-R superfamily]. PMID- 9738234 TI - [Role of reactive oxygen species in apoptosis]. PMID- 9738235 TI - [Transcription antitermination in bacteria and bacteriophages]. PMID- 9738236 TI - [Controlled expression of exogenous genes in mammalian cells]. PMID- 9738237 TI - [Regulation of human DMD gene expression. An example of multiple utilization of genetic information]. PMID- 9738238 TI - [Utilization of antisense oligonucleotide strategy in study of exposing and localization of rRNA on surface ribosomal subunits]. PMID- 9738239 TI - [Small interstitial proteoglycans--genetic differences and similarities]. PMID- 9738240 TI - [Biochemical and molecular basis of symbiotic plant and microbe interactions]. PMID- 9738241 TI - [Sense and nonsense in basic documentation]. PMID- 9738242 TI - [Therapy of bipolar affective illnesses with valproate. A review of the literature]. AB - This paper gives an overall review of the literature since 1980 on the use of valproate in bipolar affective disorders. Randomised comparative studies definitely demonstrate that valproate proves superior in comparison with placebo and equivalent in comparison with lithium in the treatment of acute manic episodes. A series of open studies indicates a reduction in frequency and intensity of manic and depressive episodes with the long-term administration of valproate suggesting efficaciousness in the prophylaxis of bipolar affective disorders. Details on dosage, side effects and potential interactions with this group of patients are presented. Valproate constitutes a further valuable and well tolerated alternative to lithium, antipsychotic agents and carbamazepine not only in acute therapy of manic episodes but also in prophylactic treatment of patients with bipolar psychosis. PMID- 9738243 TI - ["Because it concerns me"--patient-directed discharge letters]. AB - PURPOSE: Evaluation of practicability and acceptance of discharge summaries addressed directly to patients after psychiatric hospitalisation. METHODS: Over a period of 3 months 65 patients got discharge summaries addressed directly to them. Doctors and patients--4 months after discharge--were asked to evaluate this procedure. RESULTS: Both doctors' and patients' acceptance and evaluation was very positive. Argumentations for this view were the need to give and get information and the impression that this procedure can enhance confidence and trust. PMID- 9738244 TI - [Conceptual aspects in development and implementation of basic psychiatric documentation]. AB - The main conceptual prerequisites for the development and introduction of a standardised documentation system in psychiatry (in this case: psychosocial rehabilitation) are discussed. These will help to improve the acceptance of the system and optimise the quality of the data. Since the positive motivation of the staff is decisive, it is essential to meet their needs and demands. PMID- 9738245 TI - [Contribution of psychiatric departments to restructuring inpatient management: the Saxony-Anhalt example]. AB - In 1996, the available number of hospital beds for inpatients care in psychiatry, psychotherapy and psychogeriatrics in Sachsen-Anhalt amounted to approximately 0.4 beds per 1,000 inhabitants. As compared to 1992, when most beds were still concentrated in four large psychiatric hospitals, some decentralisation had been achieved. Now, 13 of 23 catchment areas have clinical facilities of their own, five of which were newly founded. About 800 of 1,346 beds and 245 of 299 day hospital places are now located outside the former large institutions. Some of the recently established facilities, however, are not yet sufficiently equipped to serve their catchment areas, with regard to appropriate size, room, staffing, internal specialisation and authorization to cope with compulsory admissions. Even in general hospitals, psychiatric departments are often separated from the main building, with the spatial distance providing some obstacle to the desired cooperation of psychiatry with other medical specialties. PMID- 9738246 TI - [Prevalence and treatment of depressive syndromes in homes for the aged. Survey of a rural catchment area]. AB - PURPOSE: The investigation aimed at assessing the prevalence of depression among residents in a nursing home. METHODS: In a rural area of Germany a representative sample was examined by means of the "Brief Assessment Interview" (BAI). RESULTS: 17% had a depressive disorder without dementia, 9% had a mild dementia syndrome and a depressive disorder; a total of 47% suffered from symptoms of dementia without depression. Most of the patients were treated by general practitioners or internists. 11% of the patients with depression were treated with antidepressants, 19% with neuroleptics, 17% with minor tranquillizers, and 17% with hypnotics. Only 20% of the depressive patients had ever been examined by a psychiatrist, 4% underwent an actual psychiatric therapy. CONCLUSION: Elderly patients with depressive disorders living in nursing homes need a more specific psychiatric treatment. PMID- 9738247 TI - [The Community Psychiatry Organization in Baden-Wurttemberg]. AB - While as a result of the report of the German Federal Government's Inquiry Commission a nationwide programme for further development of the provision of care to the mentally ill was launched, Baden-Wuerttemberg went it's own ways with its "Programme for the Development of Outpatient Psychiatric Care 1982-1986". In 1994 Baden-Wuerttemberg's concept of a "Communal Psychiatric Combine" ("Gemeindepsychiatrischer Verbund", GPV) was worked out; this concept increases in some respects the existing differences from other states. Apart from an improvement in coordination and cooperation, the GPV is intended to secure comprehensive local care provision for the chronically mentally ill. Furthermore, care provision appropriate to requirements should be made possible and quality assurance realised on all levels. In the following, Baden-Wuerttemberg's understanding of a "Communal Psychiatric Combine" is contrasted with the combine concept of the expert commission. The potential effects of the reginal care situation as well as the chances of implementation are discussed with reference to earlier experiences. PMID- 9738248 TI - [Psychotic women and their children]. AB - OBJECTIVE: The study collected information on the situation of children of psychotic mothers. METHODS: Case reports of a total of 279 female patients with schizophrenia or puerperal psychosis were analysed retrospectively. RESULTS: 128 of these patients gave birth to 218 children. Approximately 80% of the children grew up with their families, and only 8% had to be transferred to foster families or foster homes. Two case vignettes illustrate child care problems in a community psychiatric setting. CONCLUSIONS: Child care is exercised even in families where mothers suffer from psychotic illness. However, many of these mothers do not receive the help they require. PMID- 9738250 TI - [Differential diagnosis of psychotic disorders in immigrants]. AB - Several studies [5,6] on the prevalence of psychiatric disorders among migrants describe a higher rate of schizophrenic disorders. Other studies [2,3] as well as DSM-IV [1] point out that affective disorders with psychotic symptoms can be misdiagnosed as schizophrenic or schizoaffective disorders. We report on a patient admitted with the diagnosis of paranoid schizophrenia (ICD-10: F20.0), discussing differential diagnostic criteria and the problems of diagnosing along ICD-10 criteria. PMID- 9738249 TI - [Suicide over weekend leave--a seldom regarded risk factor]. AB - OBJECTIVE: The study aims at differentiating qualitative and quantitative aspects of inpatient suicide on weekdays and during the weekend. METHOD: 51 consecutive suicides in a large psychiatric hospital were analysed retrospectively. RESULTS: 14 of 51 suicides occurred during weekend leave, most of them on a Saturday or Sunday. The rate of suicides in the hospital on a Saturday or Sunday is extremely low. CONCLUSION: Weekend leave from inpatient treatment must be considered as a risk factor of its own. PMID- 9738251 TI - [Implementation of the Rodewisch Theses in East Germany--attempted analysis from the current viewpoint]. AB - 35 years ago the so-called Rodewisch theses were developed as the results of an international symposium on psychiatric rehabilitation. They were a recognised guideline in psychiatry in the GDR. During the implementation of the theses, ideological and financial limits were soon reached. Several aspects of the theses, such as decentralisation of psychiatric care, the creation of an open psychiatric system and continuity of care are still highly relevant today. PMID- 9738252 TI - [Transient psychosis in lisinopril-induced hyponatremia]. PMID- 9738253 TI - [The horn player and Ockham's razor]. PMID- 9738254 TI - [Panic disorder and/or epilepsy?]. PMID- 9738255 TI - [Factitious disorder in a mentally retarded patient in urology]. PMID- 9738258 TI - [Venous thrombosis--a diagnostic and therapeutic challenge]. AB - PURPOSE: Venous thrombosis and embolism has been the target of intensive investigation in recent years. The growing importance of mortality and morbidity due to venous thromboembolism led to new diagnostic and therapeutic tools. The clinical diagnosis may be misleading and both false-negative and false-positive diagnoses are common, when only clinical signs and symptoms are considered. METHODS: A major problem in the care of the patient is to establish the correct diagnosis. Studies show, that only Venography or Duplexsonography can clearify diagnosis. Risk factors for venous thromboembolism, propability calculations and consensus recommendations for diagnosis and therapy are presented. RESULTS: Diagnosis. Diagnosis of deep vein thrombosis must be established by Duplexultrasound and/or venography. Special situations may need further investigation by CT-scan or NMR-tomography. No single clinical sign or combination of signs give enough sensitivity and specificity. In patients with thromboembolism without clear etiology, hypercoagulability, neoplasms, compression syndroms, vascultides and other conditions have to be excluded. THERAPY: Heparins are the standard-therapy. Low-molecular weight heparins may have some benefits over standard heparins in terms of side effects and handling. Coumarin therapy can be started immediately, if no invasive procedures are necessary and have to be continued for 4-12 months, in recurred events and/or hypercoagulability even life-long. Thrombolysis is restricted to very few patients (patients under the age of 50 with large thrombosis not older than 3-7 days and no contraindications). Surgical thrombectomy is even more restricted. Compression therapy should be obligatory in all patients with thromboembolism. CONCLUSIONS: Thromboembolism is mainly a diagnostic challenge. The immediate start of the appropriate treatment may reduce embolism and thrombus formation, induce recanalisation and prevent post-thrombotic syndrom. PMID- 9738259 TI - [Preoperative imaging of varicose veins of the lower extremities. Classical pattern of insufficiency and its clinical use]. AB - Venous varicose are one of the most common diseases in industrial countries today. New surgical strategies, tailored to a patient's specific pattern of venous incompetence require more detailed preoperative imaging. METHODS: In this review of current literature we analyzed the value of ascending venography, color coded duplex sonography and descending venography for preoperative planning of varicose surgery. On the other hand, we describe variant anatomy of the superficial venous system like the different levels of escape points and perforating veins. RESULTS: Ascending venography and color coded duplex sonography are both excellent modalities for detection of reflux in the saphenofemoral and saphenopopliteal junction. The results of current literature indicate, that ascending venography is superior to color coded duplex sonography in the detection of incompetent perforators. CONCLUSIONS: Ascending venography and color coded duplex sonography provide improved information, that may be crucial for surgical planning. The high heterogeneity and broad distribution of valvular incompetence confirms the importance of detailed preoperative imaging. PMID- 9738260 TI - [Ultrasonography of veins]. AB - PURPOSE: The high incidence of acute and chronic diseases of the venous system requires the application of reliable, non-invasive, low-cost methods in diagnosis and follow-up after therapy. MATERIAL AND METHODS: Current technology, principles of examination, and results of ultrasonography of the peripheral venous systems are reviewed. RESULTS: Since the mid 1980s, compression ultrasonography (US) has been introduced in the diagnosis of deep venous thrombosis. Doppler-US methods reach the hallmarks of venous imaging, particularly since the advent of color duplex US. In thrombosis, postthrombotic syndrome, and primary varicosis, color duplex US increasingly replaces the "gold standard" of phlebography as the imaging method of choice. Venous diseases of the neck, and of the upper and lower extremities are reliably recognized by color duplex US. New areas of application of Doppler and duplex-US include examinations of the venous system in patients in intensive care units, evaluation of transplanted organs, and the demonstration of blood flow in hemodialysis shunts. CONCLUSIONS: Color duplex US is useful in most imaging investigations of the peripheral veins. In view of cost development in the medical imaging sector, however, in which ultrasonography takes a major part, critical indication for the application of Doppler- and duplex-US in the diagnosis and follow-up of venous disease is out most importance. PMID- 9738261 TI - [Contrast medium studies of the venous system]. AB - PURPOSE: To give an overview of various diagnostic techniques and indications for phlebography in different parts of the body. METHODS: Procedures of conventional phlebography of the lower and upper extremity and cavography are described and their indications in comparison to alternative techniques are discussed. The literature is reviewed with regard to specific advantages and disadvantages of the different methods. RESULTS: Conventional phlebography with iodine contrast media is still considered to be the gold standard in many regards. The diagnosis of acute and chronic thrombotic disease, venous vascular occlusions, hemodynamic malfunctions and anatomic variants of the venous system can readily be established with contrast phlebography. DISCUSSION: Main disadvantages of contrast studies of the venous system are radiation exposure and adverse effects of contrast media. Non-invasive methods such as ultrasound and MR-phlebography are becoming more and more popular and may replace venography. Other techniques such as CT-phlebography and the use of CO2 as contrast medium are under investigation. The latter can be indicated in the case of contraindications against iodine contrast media. CONCLUSION: When choosing diagnostic methods for the venous system, their sensitivity and specificity for specific diagnoses and vascular territories have to be balanced against the risks and disadvantages. PMID- 9738262 TI - [Quality assurance in phlebography of the extremities. German Societies for Angiology and Phlebology and Vascular Surgery]. AB - By interdisciplinary cooperation with the German Societies for Angiology, Phlebology and Vascular Surgery, recommendations have been worked out which address the indication of phlebography of the veins of the extremities and which define the quality standards of phlebographic techniques. The specific clinical questions in varicose syndrome acute thrombosis and postthrombotic syndrome have to be answered by employing adequate phlebographic technique, documentation, and description of relevant morphologic and functional findings. PMID- 9738263 TI - [CT-phlebography. A new method for the diagnosis of venous thrombosis of the upper and lower extremities]. AB - Spiral CT venography is a new method in vascular imaging, which is an accurate tool for the evaluation of deep venous thrombosis in the evaluation of deep venous thrombosis in the lower and upper extremity. MATERIALS AND METHODS: 102 lower extremities and 12 upper extremities were evaluated for deep vein thrombosis using spiral-CT-venography. The results were compared with findings of ascending venography, color coded duplex sonography and clinical follow up. RESULTS: Spiral CT venography of the lower extremity showed a sensitivity of 100% and a specifity of 96%. The quality of venous opacification with CT venography compared with ascending venography was superior in all venous segments. DISCUSSION: Spiral CT venography is a valuable tool for the detection of deep venous thrombosis. Advantages of the method are the reduction of the amount of contrast material necessary for opacification and the detection of perivascular soft tissue alterations. The application of CT venography is limited due to higher costs and radiation dosage. PMID- 9738264 TI - [Magnetic resonance phlebography (MRP) of the abdomen and pelvis]. AB - PURPOSE: To evaluate the image quality of magnetic resonance venography (MRV) of pelvic and abdominal veins. METHODS: A retrospective analysis of all MRV between 1993 and 1996 was conducted. A 2D-FLASH-TOF without breath hold and with arterial presaturation was used. All venous segments have been assessed for the quality of visualisation and an overall image quality was determined. 43% of our patients had malignant disease and phlebothrombosis had already been diagnosed in 64%. RESULTS: 126 examinations and 1696 venous segments were analysed. The overall quality of the examination was good, however one examination was non diagnostic because of motion artefacts. Vessels along the vertical axis were good visualised in over 90%. Most important reasons for insufficient visualisation were breathing, motion and metal artefacts as well as compression and displacement of veins due to tumour disease. CONCLUSIONS: Magnetic resonance venography without breath hold TOF technique is practical and robust for routine clinical applications. PMID- 9738265 TI - [Ultrafast MRI phlebography of the lungs]. AB - AIM: Improved detection of pulmonary-venous pathologies by imaging the pulmonary veins without arterial overlay. MATERIAL AND METHODS: Sequential 3D imaging of the pulmonary arterial and arteriovenous phase was performed with an ultrafast 3D FLASH sequence successively acquiring eight 3D data sets every 2.9 s within a single breathhold. For an 8 cm thick 3D slab an interpolated spatial resolution of about 1.4 x 1.9 x 3.3 mm could be achieved. Different protocols for contrast media dose and infusion rate were used. For selective visualization of the pulmonary veins, the pulmonary arterial phase was subtracted from a subsequent arteriovenous phase with the highest venous signal. 5 healthy volunteers, 8 patients with history of a cerebrovascular accident (CVA) of unknown etiology an suspected pulmonary-venous thrombosis and 9 patients with compression of the pulmonary vasculature by centrally growing malignancies were evaluated. RESULTS: With higher infusion rate and lower contrast media dose, arteries and veins could be better separated by their enhancement kinetics. In all cases a complete visualization of the main pulmonary veins, segmental and subsegmental veins up to the fourth order of each lung segment was achieved without any overlay of pulmonary arteries. No thrombi of the pulmonary veins were found in patients with CVA. The obstruction of pulmonary arteries and veins due to vessel compression could be selectively visualized. CONCLUSION: Ultrafast multiphase 3D-Gd-MRA is a new reliable method for selective 3D visualization of pulmonary veins. PMID- 9738266 TI - [Interventional radiology in central venous obstructions. Dilatation--stent implantation--thrombolysis]. AB - PURPOSE: Venous congestion of the superior or inferior caval system has to be considered as a medical emergency. The results of various recanalization procedures and their utility are analyzed. PATIENTS AND METHODS: 176 patients with superior and 28 with inferior caval obstruction were treated with Gianturco Z (n = 39) and Wall Stents (n = 207) respectively. Balloon venoplasty was performed prior to stent implanation. In 27 cases, local thrombolysis with urokinase was employed. RESULTS: Interventional procedures were successful in 198 and without success in 6 patients. In most patients, symptoms were relieved during or early after recanalization. No major complications were found. DISCUSSION: Balloon angioplasty with stent placement and local thrombolysis are successful in the treatment of superior and inferior caval obstruction. Self expanding Wallstents are superior to Gianturco-Z-stents. Oncologists should be made familiar with this type of treatment. PMID- 9738267 TI - [Vena cava filter. Indications, complications, clinical evaluation]. AB - INTRODUCTION: Pulmonary embolism is the third leading cause of death in the western countries. If anticoagulation fails or is contra-indicated, or if the risk for pulmonary embolism is increased for other reasons, the percutaneous implantation of a vena cava filter should be considered. METHODS: The available filters can be differentiated by the design (cone, basket, net-types), by the material, and by their removability. The rate of complications (caval thrombosis, fracture of filter) and the in vitro efficacy in trapping thrombotic clots is dependent on the specific filter type. RESULTS: In clinical practice there is no evidence for significant differences in trapping efficacy among the different filters. About 4% of all patients treated by caval filters still can have pulmonary embolism, and 1% will have a fatal outcome. Dependent on the filter type, the most common complication is caval thrombosis, in up to 25% of cases. CONCLUSION: The percutaneous implantation of caval filters can readily be performed by interventional radiologists. However, randomized clinical studies failed to clearly document efficacy of caval filters. Therefore, indication has to be considered carefully. PMID- 9738269 TI - [A rare form of elbow fracture]. PMID- 9738268 TI - [Standardized enteroclysis. Monitoring of intra-lumen pressure in 3 different patient groups]. AB - PURPOSE: Monitoring of intraluminal pressure in standardized enteroclysis. MATERIAL AND METHODS: Pressure monitoring with a double-lumen tube during fluoroscopy-guided adjustment of contrast media instillation rate due to small bowel motility and contrast media transport of 67 patients. Stratification according to patient data and findings in enteroclysis: Crohn's disease (n = 12), non-inflammatory disease (n = 35) and irritable bowel syndrome (n = 20). RESULTS: The amount, instillation period and rate of contrast media and distension media were not statistically different within the study population. Crohn's disease patients showed elevated intraluminal pressure at the end of the distension phase (47.94 +/- 10.42 cm H2O versus 38.03 +/- 10.08 and 39.55 +/- 9.74 cm H2O, respectively, P = 0.0099), as well as at the end of the examination (48.59 +/- 10.42 cm H2O versus 39.66 +/- 6.52 and 35.67 +/- 8.28 cm H2O, P = 0.0002). In comparison with both other patient groups, maximum intraluminal pressure in Crohn's disease is higher and totals 51.75 +/- 9.94 cm H2O versus 43.00 +/- 6.20 and 39.55 +/- 9.74 cm H2O, P = 0.0010. Patients with Crohn's disease require a longer instillation period of distension media (28.05 +/- 12.82 min, not statistically significant). CONCLUSION: Intraluminal pressure differs in standardized enteroclysis with fluoroscopy-guided instillation rate adjustment. Irrespective of stenosis or acute inflammation, patients with Crohn's disease show a higher intraluminal pressure compared to patients with non-inflammatory disease or irritable bowel syndrome. PMID- 9738270 TI - [Radiologists suffer the greatest loss of income. Analysis of cost structure. Analysis of the ZI (Central Institute) cost structure shows a 82% sharing of costs]. PMID- 9738272 TI - ["Nevertheless, it does move..." Negotiations on the regulations in individual health insurance plans]. PMID- 9738273 TI - [Focus on cooperation at the center of vascular surgery: radiologists and vascular surgeons seek an interdisciplinary dialogue]. PMID- 9738274 TI - [Therapy of acute myocardial infarct (1994-1996) at non-university hospitals in Switzerland (CHAMI Study)]. AB - The CHAMI study (Confederatio Helvetica Acute Myocardial Infarction) recorded the therapies administered for acute myocardial infarction in 520 consecutive patients between October 1994 and February 1996 at 10 non-academic hospitals in Switzerland. The patients in this group consisted of 363 men and 157 women with an average age of 63.2 years. The prescribed medications administered from the day of hospital admission until the day of discharge were recorded. In the acute phase, the patients were given the following therapy: thrombolytic agents 40%, i.v. nitrates 65%, i.v. beta-blockers 22%, aspirin 95%, oral beta-blockers 36%, ACE inhibitors 14%. Impressive was the lower distribution of thrombolytic agents and beta-blockers among the older patients (age > 70) (thrombolytic agents 52.1% vs 28.4%; oral beta-blockers 44.0% vs 29.1%) and in particular among women (thrombolytic agents 26.8% vs 46%; oral beta-blockers 29.3% vs 39.7%) in men. Therapy at hospital discharge consisted, inter alia, of aspirin (73%), beta blockers (54%), ACE inhibitors (3%), and lipid lowering agents (10%). The hospital mortality was 12.6%. The CHAMI study provided the participating hospitals with a quality control comparison with other participating centers and impressively demonstrated with the example of the lipid lowering agents, that the significance of secondary prophylaxis is assigned too little importance in contrast to acute therapy. PMID- 9738275 TI - [Co-trimoxazole administration: a rare cause of hypoglycemia in elderly persons]. AB - We report a case of severe hypoglycaemia following co-trimoxazole therapy. An 88 year-old woman was admitted with urinary tract infection and treated with co trimoxazole (960 mg bid). Seven days after initiation of the treatment she became comatose. Blood sugar was 1.3 mmol/l and C-peptide at the upper limit of normal range. Glucose infusion restored normal consciousness and no hypoglycaemia recurred after interruption of co-trimoxazole therapy. Advanced aged was the only risk factor identified. Other risk factors described in previous case reports are renal failure, poor nutritional state and high doses of co-trimoxazole. PMID- 9738276 TI - [How (deficient) copper causes illness]. AB - Copper is an essential cofactor in all cells. However, it remains largely unknown how cells deal with this element, which is essential yet toxic. Through the study of microbial model systems on the one hand, and the investigation of inherited diseases in copper metabolism on the other, important insights into the way cells deal with copper can be gained. Two key new elements of copper metabolism have emerged from these studies: ATP-driven copper pumps and intracellular copper transport proteins, the copper chaperones. PMID- 9738277 TI - [Physical activity and aging]. AB - About a third of the initial muscle volume is lost during human life. This decline in muscle mass is responsible for impaired muscle strength, physical frailty, falls, impaired mobility, and functional decline. The age-related loss in muscle mass accounts for the age-associated decreases in basal metabolic rate, oxygen consumption, glucose utilisation, and bone density. Several of the anatomical and physiological changes in sedentary people, commonly attributed to primary aging processes, are in fact the consequence of lack of exercise. Recent studies show that strategies to maintain muscle strength and mass among the elderly (like regular exercise training) are most important for their functional independence. PMID- 9738278 TI - [Arteriovenous fistula from the vertebral artery to the vertebral vein]. PMID- 9738279 TI - [Kinematic and neuromuscular study of the lumbar region in walking on a treadmill ergometer with and without incline]. PMID- 9738280 TI - [Joint stress in inline skating--a biomechanical study]. AB - In 8 male subjects we undertook acceleration measurements under standardised conditions at the tibia, the hip and the head. The measurements were performed on a normal flat street as well as on a bicycle route with velocities of 15 km/h, 20 km/h and 25 km/h. Additionally we documented the range of motion at the knee joint. Maximal positive acceleration at the head sensor was 10.75 m/s2 at a velocity of 25 km/h on the bicycle route. At the hip the highest measured value was documented with 15.10 m/s2 at 25 km/h also on the bicycle route. Die maximal tibia acceleration was 55.8 m/s2 at 25 km/h again on the bicycle surface. The amplitude averaged signals showed the highest suppression between tibia and hip. CONCLUSION: Compared to other athletic activities acceleration and joint load in in line-skating is only low. PMID- 9738281 TI - [Stretching--do current explanatory models suffice?]. AB - The opinion that mobility in the usual performance if the "straight leg raise" test for the evaluation of stretching techniques is subject to solety muscular limitations is critically appraised. With integration of recent results from molecular biological research and our own measurements, we can show that not only mechanical but also neurophysiological factors must be considered in the limitation of mobility. In the majority of the examined patients stretching of the ischiadic nerve seems to be responsible for restrictions in movement. The presented results cast doubt on the currently held assumptions and basic principles of stretching in therapy and sport. PMID- 9738282 TI - [Ultrasound-controlled anthropometry--on the development of a new method in asymmetry diagnosis]. AB - Recently computer-based methods allow the registration of asymmetries in skeleton axis. In comparison to classical methods the Ultrasound-Guided-Anthropometry (UGA) has been tested in regard to its quality criteria for representing body specific marks in three dimensions. In a sample of healthy subjects objectivity (n = 13), retest-reliability and internal validity (n = 26) have been determined. For analysis of system-quality a standardized cuboid phantom (side length 400 mm) has been measured. RESULTS: objectivity r = 0.93-0.98 (p < .05), retest reliability r = 0.97-0.99 (p < .05) and internal validity between both methods r = 0.92-0.97 (p < .05). The analysis of system-quality produced an error in measurement of 0.65% (0.58 +/- 1.29 mm). UGA as a screening method is not a substitute for case-history or classical anthropometry, but it offers useful parameters which facilitate decision making for further diagnostic procedures. PMID- 9738283 TI - [Traumatology and sports injuries in equestrian acrobatics in the adolescent]. AB - The objective of this study was to compile knowledge of athletic injuries and complaint patterns related specifically to equestrian acrobatics (e.a.). A 20 page standardized questionnaire was sent to 114 e.a. participants. The mean age of the participants in e.a. groups was 15 +/- 3 years, that of independent participants 21 +/- 3 years. A total of 489 injuries was reported, mainly to muscles and tendons (35%), skin (33%) and joints and ligaments (25%). Bone injuries (6%) and head injuries (2%) were infrequent. Analysis of the localization showed that the head-face-neck region was involved in the injury in 3.9%, the torso in 5.5%, the upper extremities in 28%, pelvis and hip region in 3.9%, and the lower extremities in 52%. More than half the injuries were categorized in severity grade I (55%) (not requiring medical attention), 25% were grade II (single medical treatment), 15% grade III (several outpatient medical treatments) and 5% grade IV (requiring hospitalization). Nearly half of the injuries to the lower extremities resulted from jumping exercises, while the cause in upper-extremity injuries was mostly falling (37%). The importance of fall training and limitation of difficulty as well as the number and height of the jumps is discussed. Regular medical examination and improved education of the trainers are demanded. PMID- 9738284 TI - [Incidence of injuries in parachuting]. AB - During the German championships in parachuting 78 paratroopers were asked about acute injuries and chronic pain using a questionnaire. A total amount of 131 injuries was described. These were evaluated in terms of dimension and localisation. Upper and lower parts of the body were injured with a comparable frequency. Bruises (42%), fractures (19%), sprainings (16%) and dislocations (10%) were most often described. The overall injury rate according to the total number of descents (0.09%) was lower than that reported by previous literature. Therefore it can be concluded that parachuting for experienced jumpers is less dangerous than assumed until today. PMID- 9738285 TI - [Fracture of the tarsal navicular bone in childhood--therapy and functional follow-up]. AB - Using two patients as examples we describe the therapy and results of children with fractures of the tarsale os navicular. A conservative therapy is striven generally for slight dislocated fractures without interruption in the articular facet. At dislocated fractures, fractures with luxation or interruption of the articular facet an open reposition and retention with K-wire or screw is recommendable. Our functional check-up by means of dynamic pedography, the measurement of pressure, force and time under the sole locally and time wise dissolved, show subjectively not visible standard deviations, which normalized during a period by about 1 1/2 years. Differential diagnosis are aseptic bone necrosis as well as additional apophysis. PMID- 9738286 TI - [Sudeck disease--pathology, clinical aspects and therapy]. AB - In our opinion the etiology of Sudeck's disease (acute reflex bone atrophy) plays a decisive role in therapeutic planning. The therapy is based on clinical and radiological findings. Physiotherapy addresses the symptom complex of pain, hyperemia, edema formation, and limitations of movement which act in a vicious circle and its intensity is modified according to the prevailing clinical and possibly also radiological findings. A strict coupling of the therapy to a classification according to stage is not recommended. Pharmacological therapy is merely a supporting element and focuses on the sympathetic overexcitability. The best therapy for Sudeck's disease is prophylaxis. Interventions collected under the general term early functional mobilization are, especially after surgical measures, a major factor in the avoidance of neurovegetative dysregulation in the sense of sympathetic reflex dystrophy. PMID- 9738287 TI - [Treosulfan displays cytotoxic effect on spheroids of primary cell cultures of renal cell carcinoma independent of p-glycoprotein expression]. AB - Therapy of advanced renal cell carcinoma remains difficult. New therapeutic schemes besides cytokine treatment should be evaluated. The following study analyzes the in vitro toxicity of treosulfan on spheroids of 8 primary cultures of renal cell carcinoma cells. these data were compared to the toxicity of vinblastine. All investigations were performed in regard to the P-glycoprotein (Pgp) expression of the cells, which is one of the main causes of multidrug resistance. Four Pgp positive and four Pgp negative spheroids were incubated with the drugs in increasing doses. Toxicity was measured using the MTT toxicity assay as well as trypan blue exclusion. Significantly higher toxicity of treosulfan compared to vinblastine could be demonstrated. In addition, the effects of treosulfan were not related to Pgp expression. These results are encouraging and a phase II study analyzing the efficacy of treosulfan in patients with advanced renal cell carcinoma has been initiated in our institution. PMID- 9738288 TI - [Results of ureterocystoneostomy for inner urinary diversion in locally advanced prostate carcinoma]. AB - Ureterneoimplantation (unilateral in 6 cases) was performed as palliative urinary diversion in 8 patients (age 64-81 years) due to locally advanced prostate cancer and bilateral ureteral obstruction (serum creatinine 2.1 to 9.8 mg. per dl.) between 1991 and 1995. In these cases the application of a double-J-catheter had failed or a percutaneous nephrostomy was refused. Postoperative time of survival (237 days, 2 patients still living for 20 and 21 months after therapy), mortality (1 of 8 patients), morbidity and time to hospital discharge (26 days) are compared to the results of the published retrospective investigations concerning percutaneous nephrostomy. The opportunity of a natural micturition without external urinary diversion could be gained for a longer period of time (5 and 20 months) in 2 of 3 patients. The other patients with in situ double-j-catheters were drained sufficiently by a suprapubic cystostomy (serum creatinine postoperatively 1.3 to 2.0 mg. per dl.). Bilateral ureterocystoneostomy being more invasive than unilateral diversion showed no benefits and was no more performed since 1991. Uretemeoimplantation with comparable postoperative results to percutaneus nephrostomy seems to be a sufficient therapeutic possibility in patients with natural micturition, repeated catheter complications, refusal or failure of alternative urinary diversion. PMID- 9738289 TI - [Metalloproteinases (MMP-1, MMP-3) and their inhibitors (TIMP) in blood plasma of patients with prostate carcinoma]. AB - Matrix metalloproteinases (MMP) and their tissue inhibitors (TIMP) are involved in important processes of tumor invasion and metastasis. In the study presented, matrix metalloproteinase 1 (MMP1) and 3 (MMP3), the tissue inhibitor of metalloproteinase 1 (TIMP1) and the complex MMP1/TIMP1 were measured by ELISA tests specific for these proteins in blood plasma. These components have been investigated in prostate cancer patients (PCa) with metastases (n = 18; T2, 3, 4 pN1, 2M1), prostate cancer patients without metastases (n = 29; T2, 3 pNOMO), patients with benign prostate hyperplasia (BPH; n = 29) and in healthy men (n = 35). Mean values of MMP1 and of the complex MMP1/TIMP1 were not different among the four groups. The mean values of MMP3 and especially TIMP1 were significantly higher in prostate cancer patients with metastases compared with controls, BPH patients and prostate cancer patients without metastases. Ten of these 18 patients had TIMP1 levels higher than the upper reference limit. TIMP1 concentrations correlate to the tumor stage but not to the tumor grade. These results indicate, that TIMP1 could be an potential marker for metastases in prostate cancer patients. PMID- 9738290 TI - [Primary paraurethral malignant mixed tumor (carcinosarcoma) of the vagina. A case report]. AB - In total malignant mixed tumors (carcinosarcomas) represent a very rare gynecological neoplasm. In accordance with the embryological genesis primary localizations are with descending probability the uterus, the ovarians and finally the tubes. We report on a 55 year old female patient, with a paraurethral mass, which was primarily diagnosed in 1994. Histopathological evaluation of a transvaginal biopsy did not demonstrate evidence of malignancy at that time. After painful enlargement the paraurethral mass was resected surgically and histopathological evaluation revealed a primary paraurethral malignant mixed tumor (carcinosarcoma). Postoperatively, the patient underwent percutaneous (46.4 Gy) and intracavitary (2 x 6 Gy) radiation. Twelve months postoperatively there is no evidence of disease. PMID- 9738291 TI - [Telomerase in tumors: facts and perspectives]. AB - Recent reports have indicated the ribonucleoprotein enzyme telomerase to play an important role in tumorigenesis. Activation of this enzyme is known to prevent progressive shortening of the end of the chromosomes, or telomeres, and hence to be critical in maintaining chromosomal integrity. The telomerase expressing cells require immortality. Supported by recent findings which suggest that telomerase activity is expressed in virtually all cancers but not in normal tissues, except those of the germline, hope grew up toward a potentially important new therapeutic target in the fight against cancer. An emerging hypothesis is that the inactivation of telomerase results in the death of immortal cells. If telomerase activation represents a tumor-specific feature, (gene) therapeutic applications would become most promising in regard to an effective anticancer therapy, possibly with limited side-effects. However, most recent studies report on telomerase activity to be also expressed in normal, non-neoplastic tissues as well as in non-neoplastic hyperproliferative lesions. The questions whether telomerase activity is tumor-specific or associated with (physiologic or pathologic) hyperproliferation is intriguing and remains to be clarified. PMID- 9738292 TI - [Technique of multiple organ procurement. Is involvement of urology indicated?]. AB - Today the organ donor operation in brain dead donors is mostly projected as multiple organ procurement (MOP). Not only the kidneys, but heart and liver or additionally lungs and pancreas are removed in MOP. A good synchronization between the collaborating transplantation groups (thoracic and abdominal surgeon, urologist) is essential to prevent loss of an organ because of technical problems. In Germany urologists perform more than 40% of kidney transplantations. These urologic institutions perform the cadaver kidney retrievals and participate on MOP. If possible, an urologist experienced in renal transplantation should contribute to the care for quality of the kidneys. Since 1987 the transplant center of Bremen obtained 390 donor registrations. 202 organ donor operations have been performed (106 MOP and 96 exclusive kidney retrievals). 398 donor kidneys have been collected and 382 could be transplanted. PMID- 9738293 TI - [Diseases of the glandulae bulbourethrales Cowperi in the man]. AB - The history of Cowper Gland is described. Next the clinical, radiological and endoscopical findings in 172 patients (age 18 to 82) are evaluated with regard to the importance of these alterations in clinical practice. There was no tumor. In three cases, only the bulbourethral gland showed inflammation. Out of 169 syringoceles 9 were closed (3 perineal, 6 bulbar). Only one case of the urethral (bulbar) type of closed syringocele showed obstruction. There were 56 patients with various forms of dilatation and opening into the urethra. Only one case showed obstruction. Thirty-one out of 104 patients with so-called simple syringocele had mild dilatation of the duct, mainly normal gland and mild symptoms. Alterations of the urethral wall as well as clinical symptoms were noted in the remaining 73 men with irregular ducts and alterations of the gland; all of them had local urethral signs; in 42 there were extensive urethral strictures. PMID- 9738294 TI - [Transurethral alprostadil administration with MUSE ("Medicated Urethral System for Erection"). Current overview and personal experiences]. AB - An analysis of the presently available results concerning transurethral application of Alprostadil with MUSE (Medicated Urethral System for Erection) up to 1000 micrograms indicates a 20-30% lower efficacy if compared to 20 micrograms i.c. injected Alprostadil. Whereas in prospective home-treatment trials only each second MUSE-application was successful in responders 87%-94% of the administrations in self-injection therapy resulted in successful coitus. In an own comparative trial in 73 pts the success rates after MUSE up to 1000 micrograms were 48% compared to 71% after i.c. Alprostadil. Reported side-effects of MUSE in the literature were: Hypotension 3-8%, syncopes 0.4%, penile/urethral pain 29%, urethral bleeding 5%, vaginal irritations 3%, priapisms < 0.1%. After MUSE-applikation the average Alprostadil contents of the ejaculate increased 40%. Whereas in prospective long-term studies of self-injection therapy with Alprostadil the risk of persistent fibrotic alterations of the penis varied between 5-7%, the risk of penile fibrosis after MUSE can not be finally estimated. Also the potential risk for urethral strictures after MUSE is presently not foreseable. The advantage of the technically easy use in confronted with a considerably lower efficacy. Therefore self-injection therapy must be further on considered the "golden standard" in Alprostadil administration. PMID- 9738295 TI - [Percutaneous bladder neck suspension with osseous fixation. Surgical technique and initial results in 26 patients]. AB - A modification of percutaneous needle suspension using a bone anchor system for fixation of the suture to the pubic bone is presented. After a follow-up of 3-11 months (mean 6,8) the postoperative success-rate in 26 patients is 73.1% (n = 19). A removal of bone anchor and suture was necessary in 2 patients because of bacterial infection or soft tissue granuloma. We can not reinforce the very optimistic results of the first reports published in the recent years. Therefore it is still questionable whether percutaneous needle bladder neck suspension should be a first line procedure for the treatment of female urinary stress incontinence. PMID- 9738296 TI - [Improving the specificity of PSA's in the diagnostic gray zone 4-10 ng/ml by human glandular kallikrein]. PMID- 9738297 TI - [Register study for treatment of brain metastases of malignant testicular tumors]. PMID- 9738298 TI - [Do close but negative margins in radical prostatectomy specimens increase the risk of postoperative progression?]. PMID- 9738299 TI - [External electrostimulation in therapy of urinary incontinence]. PMID- 9738300 TI - [Colorectal carcinoma. Prevention and early detection]. PMID- 9738301 TI - [Guidelines for diagnosis of bladder cancer. German Society of Urology]. PMID- 9738302 TI - Families with multiple cases of gluten-sensitive enteropathy. AB - Early detection of oligosymptomatic gluten-sensitive enteropathy (GSE) may contribute to the prevention of late complications, such as malignancy. Family members of known GSE patients are at higher risk of being affected. To evaluate the frequency and clinical significance of multiple occurrence, we routinely offered an antiendomysium antibody (EmA)-based non-invasive screening to affected families. Among 997 family members of 396 GSE patients, we identified 89 subjects with EmA positivity and/or severe jejunal villous atrophy. In 83 cases GSE has been verified, four patients refused the biopsy and two subjects are under further observation for latent celiac disease. Prevalence of GSE was 8.5% (80/943) among the first-degree relatives, with significantly higher values in the siblings (13.8%) and offsprings (12.0%) than in the parents (4.2%) of the probands (p < 0.001). In 55 families (13.9% of the families studied) two, in ten families (2.5%) three, in one family four and in one other family six members were affected. Combinations of the clinical presentations of index and screening detected cases were highly variable, with a high percentage of silent and atypical forms in the relatives. GSE cases presenting both with and without dermatitis herpetiformis occurred in 15 families. Six GSE cases with atypical or mild dermatitis herpetiformis were detected in consequence of the screening. CONCLUSIONS: EmA-assisted family screening resulted in the detection of a clinically significant number of additional GSE patients. PMID- 9738303 TI - [ETICS Study: Empirical therapy of idiopathic chronic singultus]. AB - Idiopathic chronic singultus (ICS) describes recurring episodes of persistent hiccuping lasting longer than an arbitrary time limit (e.g. one month) for which no organic cause or consistently effective treatment can be found. It has been suggested that ICS may result from chronic stimulation of a supraspinal "hiccup center" by impulses originating from receptors in the gastrointestinaltract. This hypotesis implies the possibility of treating ICS by reducing gastric acid (via omeprazole), facilitating gastric emptying (via cisapride), or suppressing of the "hiccup centre" (via GABA-ergic offects of baclofen or gabapentin). 29 patients (28 male, one female; age 71 +/- 10 years) suffering from ICS for four to 564 months were treated with a combination of cisapride (30 mg/d), omeprazole (20 mg/d) and baclofen (45 mg/d) (COB). The patients rated the severity of hiccuping on a subjective assessment scale (SAS) that ranged from 0 (= no hiccuping) to 10 (= unbearable hiccuping). Hiccuping ceased in 38% (11/29) of the treated patients and decreased in severity in an additional 24% (7/29). Mean SAS-scores following 20 to 24 weeks of therapy (3.7 +/- 3.4) were significantly lower compared to before therapy (8.8 +/- 1.3) (Mann-Whitney rank order test [p < 0.02]). In the patients that failed to respond to COB, gabapentin (1.200 mg/d) was substituted for baclofen. Hiccuping ceased in one and improved in two of these ten subjects. We conclude that COB is an effective empirical therapy in the majority of patients with ICS. It may be useful to substitute gabapentin for baclofen in those not responding to COB. PMID- 9738305 TI - [Enteroparesis caused by progressive fibrosis of the muscularis propria]. AB - We would like to present the case of a 54-year-old patient who was admitted to the hospital with complaints of recurrent vomitus after food intake and associated weight loss. Endoscopy. barium X-ray of the small bowel and scintigraphy showed enteroparesis. Full-thickness biopsies achieved by explorative laparotomy revealed the pathological changes. The inner circular muscular layer was atrophic and replaced by fibrosis. The pathological findings were consistened with either visceral myopathy or isolated intestinal progressive systemic sclerosis. A sporadic nonfamilial type of visceral myopathy seems to be the most likely diagnosis in this case. Progressive systemic sclerosis seems to be unlikely as the patient presented with isolated gastrointestinal involvement and lack of appropriate autoantibodies. The diseases progression made an enteral supplementation impossible. Therefore a parenteral nutrition was started, which was characterized by a complicated clinical course. Persisting gastrointestinal complaints, recurrent port infections and lack of perspectives led to patient's diminished motivation for adequate sterile use of the port-system. This case report shows that besides the use of modern port-systems and antibiotics the psychological situation of patients treated with total parenteral nutrition is of great interest for optimal patient care. PMID- 9738304 TI - [An unusual course of low-malignancy non-Hodgkin lymphoma of the stomach]. AB - We report on a 51-year-old women, who suffered from a low-grade lymphoma of the stomach (MALT-lymphoma) and underwent subtotal gastrectomy in 1991. Two years later she developed a relapse of her MALT-lymphoma. She was treated with two Helicobacter pylori eradication therapies which led to complets remission of the lymphoma. In 1994 she developed a high-grade conchal lymphoma and underwent conchotomy. No moleculargenetic evidence of any relationship between the two lymphomas was found. PMID- 9738306 TI - [Extrahepatic manifestations of hepatitis C infection]. AB - The hepatitis C virus is associated with various extrahepatic manifestations. It is unequivocal that hepatitis C is associated with mixed cryoglobulinemia, membranoproliferative glomerulonephritis, and in Southern Europe to some extend also with porphyria cutanea tarda. The rare combination of hepatitis C and panarteriitis nodosa is still questionable. The Sicca syndrome also seems to be associated with hepatitis C virus, but this is not the typical Sjogren syndrome. It is still unclear whether the rare combination of hepatitis C with aplastic enemia has pathogenetic aspects. Although an epidemiological association of hepatitis C with lichen planus, lymphoma, neuropathies and other diseases has been observed, the etiological role and the pathogenetic involvement of the hepatitis C infection remains unclear. Furthermore, the question whether these extrahepatic diseases observed are autoimmune diseases remains open. Concerning the practical approach in a clinical setting, it has to be pointed out that with these diseases a hepatitis C infection has to be considered and testing for hepatitis C antibodies and, if positive, hepatitis C-RNA is indicated. If there will be any evidence of an etiological association of a replicative hepatitis C infection and the mentioned extrahepatic diseases antiviral medication should be discussed. PMID- 9738307 TI - [Antibody diagnosis iin sprue/celiac disease]. AB - The clinical appearance of celiac disease is diverse, often times uncharacteristic and may therefore be very difficult to diagnose. Furthermore, severe inflammation of the small bowel can be present without gastrointestinal symptoms. It is estimated that most subjects with celiac disease are not diagnosed. Yet, early diagnose is desirable, since celiac disease causes growth retardation in untreated children and is potentially cancer disposing. Thus, all patients should adhere to a strict gluten-free diet. The role of antibody testing and small bowel biopsy in celiac disease will be discussed. In the future, the development of new tests may allow screening of large populations. PMID- 9738308 TI - 3D-ultrasound in imaging, diagnosis and follow-up of an atypical hydatid cyst. AB - Abdominal ultrasonography is the procedure of choice to diagnose hydatid cysts caused by Echinococcus granulosus. Recently three-dimensional ultrasonography has become available for clinical application. We report a case of an atypical seronegative hydatid disease, in which the additional use of 3D-sonography improved the sonographic diagnosis, which was confirmed by a fine needle biopsie complicated by an anaphylactic reaction. In addition the potential advantage of 3D-ultrasonography in diagnosis and follow-up hydatid disease will be discussed, especially in the context of new alternative therapeutic options like chemotherapy with benzimidazoles or the percutaneous drainage by the PAIR procedure (puncture-aspiration-injection-re-aspiration). PMID- 9738309 TI - [Does anti-estrogen therapy after estrogen receptor analysis have value in palliative therapy of hepatocellular carcinoma?]. PMID- 9738310 TI - [Routine computerized tomography in suspected acute appendicitis?]. PMID- 9738311 TI - [Is gastrin trophic? Gastrin/ gastrin receptor "knockout" mice confirm the old hypothesis]. PMID- 9738312 TI - Dimensions of subjective uncertainty in social identification and minimal intergroup discrimination. AB - Social categorization under minimal group conditions reliably produces intergroup discrimination. It is proposed that this might be because the minimal group paradigm engenders high levels of subjective uncertainty among participants, which causes them to use the categorization to define self and thus identify with the minimal group. Uncertainty is generally an aversive state which may be resolved by identification (Hogg, 1996; Hogg & Abrams, 1993). Thus, people may identify with social categories (and express discrimination) when identification resolves uncertainty. To investigate this idea a standard minimal group experiment was conducted, in which the three independent variables of categorizations, task uncertainty and situational uncertainty were manipulated in a 2 x 2 x 2 factorial between-subjects design. Point distribution strategies were measured along with in-group identification, self-esteem, social awareness and, at three occasions, uncertainty about the task and situation. As predicted, under conditions of high task or situational uncertainty, categorized participants identified more with their minimal in-group and exhibited more intergroup discrimination than other participants. There was also some evidence that identification reduced uncertainty. There was only partial support for the mediational role of identification, and similarly, for self-esteem as a derivative motive. These findings are interpreted as supporting an uncertainty reduction model of social identification and group motivation. PMID- 9738313 TI - Video game playing and its relations with aggressive and prosocial behaviour. AB - In this study of 278 children from the seventh and eighth grade of five elementary schools in Enschede, The Netherlands, the relationship between the amount of time children spent on playing video games and aggressive as well as prosocial behaviour was investigated. In addition, the relationship between the preference for aggressive video games and aggressive and prosocial behaviour was studied. No significant relationship was found between video game use in general and aggressive behaviour, but a significant negative relationship with prosocial behaviour was supported. However, separate analyses for boys and girls did not reveal this relationship. More consistent results were found for the preference for aggressive video games: children, especially boys, who preferred aggressive video games were more aggressive and showed less prosocial behaviour than those with a low preference for these games. Further analyses showed that children who preferred playing aggressive video games tended to be less intelligent. PMID- 9738314 TI - [Surgery in day hospital: scrotal surgery]. AB - Actually scrotal ambulatory surgery represents a necessity for the surgeon because of the more and more greatest request to admittance and because of the impossibility to admit patients with less severe diseases. Ambulatory treatments allows psychological advantage, less discomfort, less hospital complications for the patients and a reduction of sanitary expense. Light general anaesthesia with concomitant administration of local anaesthesia is performed in our Institute. Surgical treatment must be short, with low haemorragic risk, scarcely algogenic and aseptic. Since January 1994 till December 1997, 484 patients underwent scrotal ambulatory surgery in our Institute. We reported one cardiac black and two lypothymia attacks associated with anaesthesia. Therefore combined general anaesthesia results a valid technique but unvoid of complications. Ambulatory surgery for scrotal surgery represents a sure advantage for patients and for the sanitary expense. PMID- 9738315 TI - [Urethral and prostatic urologic endoscopic surgery in day hospital]. AB - We evaluated urethral and prostatic urological endoscopical surgery in Day Hospital, following rules proposed by Veneto Region in 1996. We made surgical treatments for urethral strictures, BPH obstruction, and sclerosis of the bladder neck in 44 patients (age: 67-84). Laser therapy and classical surgical techniques were used. Our results were good: complete resolution of obstruction, lack of bleeding, early catheter removal, and acceptable costs. PMID- 9738316 TI - [The basquet in day hospital]. AB - We use the basket as the first choice method to treat the pelvic ureteral stones, according to the standard technique. In the last two years ('96, '97) the basket has been used in 49 cases (27 and 22 respectively) with pelvic ureteral stone; 4 of them were treated also by ESWL. So, 45 Pz have been treated only by basket and the success was obtained in 43 cases (95.5%). The 74% (32/43) of the cases were completed in a single step procedure and the 72% (23/32) of them have been discharged on the subsequent morning. Considering these results it is possible to perform the technique during a day hospital on condition that some parameters are respected: little size of the stone, seat near the bladder, easy insertion of the basket and female patient preferably. In any case we have to inform the patient about the possibility to proceed with a delayed extraction. PMID- 9738317 TI - [Echoguided percutaneous nephrostomy]. AB - Percutaneous US guided nephrostomy is the simplest and most direct technique to drain an obstructed kidney. The indications are included in two groups: temporary drainage and permanent drainage; the former is indicated in the non endoscopically superable ureteral obstruction, in pyonephrosis, in pregnant women and in transplanted kidneys (due to the easier access), the latter is exclusively reserved to neoplastic obstructions. The only real contraindication to the method, besides a documented allergy to local anaesthetics, is represented by a severe coagulopathy. Positioning techniques are the "one shot" technique, in which dilation and positioning are synchronous (it can avoid fluoroscopy but it is more traumatic) and angiographic derived Seldinger's technique, that utilizes fluoroscopy and an instrumentation including a guidewire and a set of Amplatz dilators. Complications are due to the access route; the choice of an intercostal access is always inadvisable, due to the risk of pneumothorax or pulmonary injury; the most frequent complications are vascular (hemorrhage, retroperitoneal hematoma) and usually well controlled; more severe lesions (renal artery laceration and arteriovenous fistula) may require intervention or embolization, but the incidence of nephrectomies due to vascular injury accounts for one per thousand. PMID- 9738318 TI - [Urologic laparoscopy in day hospital]. AB - Thanks to the less invasiveness that is the limitation of the surgical trauma, the utilization of laparoscopic techniques for managing a wide range of urological disorders has vastly expanded over the past five years. The advantages of less postoperative pain, shorter convalescence and decreased analgesic use collide with the high costs and the necessity of a proper training. Few cases are reported in the literature about urological laparoscopic procedures performed on an outpatient basis. This is due to the requirement of the general anesthesia in most cases with prolonged operative and anesthesiological times. Urological laparoscopic procedures are applicable on an outpatient basis only if the length of the surgery is less than two hours and the patients are able to collaborate and without concurrent medical problems. Therefore we think that the urological laparoscopic diagnostic procedures feasible on an outpatient basis could be: pelvic lymphadenectomy, identification of intra-abdominal testis and intersex, renal biopsy. Varicocelectomy, intra-abdominal orchiectomy, renal cyst resection, pelvic lymphocelectomy and bladder neck suspension can be performed as same-day surgery. We emphasize that the appropriate patient selection is critical: the ability of the patient and his family to care for the patient at home following surgery is of paramount importance in the decision to proceed with outpatient surgery. The availability of appropriately trained and experienced doctors and nurses are crucial for the potential postoperative complications. PMID- 9738319 TI - [Male and female urinary incontinence: treatment in day surgery]. AB - Incontinence isn't itself a disease but the feature of possible urinary tract alterations or outside of it. Incontinence is frequent above all in the elderly but it can be on charge of both sexes at every age. In Italy, according to recent evaluations, people affected with this disease would be more than 4 millions. Incontinence is therefore an important failure for its health aspects but also for economic and social ones. The problem is to evaluate if incontinence can't be prevented and as consequence needs only an assistance management, or it can be considered a preventable disease able to be cured, as we deeply believe, suggested also by the positive results of new therapeutical procedures, in association with traditional surgery and rehabilitation such as injectables or mini-invasive quick operations such as colpocleisis or percutaneous vaginal colposuspension (PVC), matters of this presentation and always performed according to correct diagnosis and indication. Bovine dermal collagen highly purified, poorly viscous and easily injectable, despite traditional rehabilitation and surgery, is a further procedure, endoscopic and minimally invasive to treat stress incontinence. Collagen is employed to perform a bladder neck plasty, increasing urethrosphincterial competence, to obtain continence without the creation of an obstruction. Genital prolapse, that is hysterocolpocele or simple vaginal vault prolapse, has course in high proportion (37%) in elderly (after 80 years). Surgical management of severe failures of continence and often also of the voiding function, such as: hyscuria with vesicoureteral reflux, obstinate constipation related to severe genital prolapse with allied rectocele is often hardly performed in elderly owing to the age and general health conditions: colpoclesis is a vaginal surgical approach that can be easily performed by the urologist too, it is an effective alternative to permanent catheterization or maxipad to be offered to the patient to improve her quality of life. In between the above maintained procedures takes place the percutaneous vaginal colposuspension (PVC). It is an original technique made up in our Institute to treat incontinence by the bladder neck resuspension to Cooper ligament according to a complete miniinvasive retropubic tension free transvaginal colposuspension, in local anaesthesia and complementary light narcosis in Day Surgery. Urinary incontinence is today a disturbance easy to be cured thanks to injectables and to miniinvasive surgical procedures as reported in this presentation concerning the most advanced approaches to its management. PMID- 9738320 TI - [Extracorporeal SWL in the treatment of reno-ureteral calculosis in day hospital]. AB - Eighteen years after the first clinical shock wave lithotripsy (SWL), no doubt remains as to its therapeutic efficacy in ureterorenal lithiasis. The advent of lithotriptors with a large shock wave energy range and integration of both ultrasound and radiologic imaging equipment at the shock wave source has meant that outpatients treatment of urolithiasis is now feasible in a good proportion of cases. In our lithotripsy center, from January 1995 to August 1996, 208 out of 310 patients who underwent SWL treatment for renal and ureteral stones, were outpatients. Pretreatment manoeuvres were performed in 10.6% of the patients. No major complications occurred during the treatment. Only three patients (1.4%) were admitted to hospital because of fever, colics or perirenal haematoma in the first two days after SWL therapy. The stone free rate was 67 and 84% respectively one and three months after treatment. In our experience, the possibility of performing SWL treatments without anesthesia and even analgosedation, the absence of complications and the high success rate, make outpatient treatment of urolithiasis safe and suitable in a large number of patients. PMID- 9738321 TI - [Endoscopic surgery in day hospital using bladder and urethral anesthesia with EMDA]. AB - PURPOSE: To assess the suitability of EMDA local anaesthesia for invasive procedures on lower urinary tract in one day surgery treatment. PATIENTS AND METHODS: The deep penetration of lignocaine into the bladder wall was attained by catheters, electrodes and electric current generators using revised electrochemical principles. Since 1994 203 patients with transitional cell carcinoma of the bladder underwent TURBT and 70 patients with bladder neck or prostatic obstruction underwent TURP, TUIP, TULAP; 20 patients underwent miscellaneous procedures: in 34 patients the procedures were combined. The patients' age was within 20 and 90 years (mean age 67.3). The procedures were performed in a single small endoscopic theatre annexed to the Urology Ward. A standard rigid resectoscope was used as well as a standard electrocautery (360 kHz) or a mega frequency low temperature Vesalius generator (4 MHz). Most patients received a premedication and some of them a light sedation when necessary, but all of them were fully conscious and able to complete an assessment using a simple pain scale. RESULTS: 8 out of 273 patients (3%) considered pain intolerable and other 11 (4%) reported painful but tolerable sensation, and the remaining 254 patients referred absent or minimal discomfort. Most of the patients were able to walk back to their room and go home in the evening. Those who had no chance of going back home were admitted for the night as well as those who had no assistance at all at home or those who had high probability of haemorrhage. Side effects were minimal and not related to local anesthesia: the serum lignocaine levels measured in 4 patients were innocuous. All patients experienced some tingling and reddening at the skin site of the dispersive electrode fading in a few hours. CONCLUSIONS: Local anaesthesia by using EMDA proved to be effective for most invasive endoscopic procedures in the lower urinary tract and suitable for patients undergoing day hospital surgery. PMID- 9738322 TI - [Ingeniousness and economy, basic ingredients in the circulatory adaptation to the vertical position]. PMID- 9738323 TI - Passive smoking and coronary heart disease. PMID- 9738324 TI - [Control of the cardiovascular system during sleep]. PMID- 9738325 TI - [Interventional hemodynamics in pediatric cardiology]. PMID- 9738326 TI - Coronary artery plaque dimension and rupture. PMID- 9738327 TI - ["Anatomical M-mode": a new echocardiographic technique potentially useful in the quantitative study of the left ventricle]. PMID- 9738328 TI - [Stress echocardiography in the study of the warm-up phenomenon]. AB - To date, the "warm-up" phenomenon in patients has been evaluated by ECG and symptom analysis. We investigated the warm-up phenomenon with supine bicycle stress echocardiography in patients with coronary artery disease documented by angiography and positive stress echocardiography. Sixteen coronary artery disease patients (54 +/- 9 years), who were off treatment throughout the study, were enrolled. Each of them underwent two consecutive exercise tests (25 W/2 min) with a 10-min recovery to reestablish baseline conditions. At the end of each stage of exercise and at peak exercise, when wall motion abnormalities (WMA), 1 mm ST depression and angina occurred, and at each minute, for the first 6 min of recovery, a 12-lead ECG was recorded and rate-pressure product was calculated. Time of onset and duration of 1 mm ST depression, WMA and angina, were also determined. Peak WMA, peak wall motion score index, duration of exercise and severity of angina were also evaluated. Exercise time duration and peak rate pressure product were greater during the second than the first test (p = 0.02, p = 0.03 respectively); the second test also showed a longer delay of the onset of 1 mm ST depression and WMA (p = 0.01, p = 0.01 respectively) and higher rate- pressure product values (p = 0.04, p = 0.03 respectively). On the contrary, wall motion score index during the first and the second test was similar. Time to angina onset was longer during the second test (p = 0.03); the recovery period of ST depression and WMA was shorter during the second test (p = 0.02). In conclusion, these preliminary data show that patients tolerated the second period of ischemia better than the first, consistent with the presence of the warm-up phenomenon. However, the similarity of values of wall motion score index and WMA did not support a reduction in the ischemic area during the second test. This is in contrast with a possible modification of myocardial metabolism which typically underlies the ischemic preconditioning. PMID- 9738329 TI - Antithrombotic therapy after coronary stent placement. AB - Subacute stent thrombosis and hemorrhagic complications due to intensive anticoagulant therapy limit the clinical benefit of coronary stenting. Antithrombotic therapy after coronary stent placement has not been standardized yet. From January 1994 to December 1995 a total of 338 Palmaz-Schatz stents were implanted in 285 patients. Procedural success rate was 98.8%. In the initial period, after stent placement, patients were treated with acetylsalicylic acid (ASA) and warfarin (135 patients, Group A), while subsequently, according to the results of other studies, patients were treated with ASA plus ticlopidine (146 patients, Group B). Two hours after sheath removal, Group A patients were treated with intravenous heparin until therapeutic INR (2.5-3.5) was reached; warfarin was stopped 3 months later. In Group B patients 2 hours after sheath removal a treatment with subcutaneous heparin 25,000 IU/die plus ticlopidine 500 mg/die was started. Subcutaneous heparin was maintained until hospital discharge, ticlopidine was stopped after 1 month and ASA was maintained indefinitely. There were no significant differences in baseline characteristics between the two groups. Most patients had unstable angina and in the majority of cases the stent was implanted due to intimal dissection after balloon dilation. Eleven patients had subacute thrombosis of the stent (3.9%): 9 patients were in Group A (6%) and 2 patients were in Group B (1.3%; p = 0.04). Seven patients (6 in Group A, 1 in Group B) were treated with emergency coronary angioplasty and 3 (2 in Group A, 1 in Group B) with coronary bypass; nevertheless 7 patients (6 in Group A, 1 in Group B) had an acute myocardial infarction. Eight patients (6 in Group A, 2 in Group B) had major bleeding due to a large groin hematoma requiring blood transfusion or vascular surgery. In conclusion, after coronary stenting antithrombotic therapy with ASA plus ticlopidine, as compared with anticoagulant therapy, reduces the incidence of both cardiac events and hemorrhagic complications. PMID- 9738330 TI - [Safety, feasibility and efficacy of a new single-wire stent in the treatment of complex coronary lesions: the angiostent]. AB - The angiostent is a single wire, flexible, highly radiopaque, balloon expandable stent. To evaluate the feasibility and safety of the deployment of this new stent, we report the clinical and procedural results of 70 procedures performed in 51 native coronary arteries of 48 patients, with objective evidence of ischemia. The target lesion was located in the left anterior descending artery in 18 (36%) cases, in the circumflex artery in 16 (31%) cases and in the right coronary artery in 17 (33%) cases. Mean reference vessel diameter was 3.2 +/- 0.4 mm and the minimal luminal diameter was 0.4 +/- 0.3 mm, with a mean diameter stenosis of 86 +/- 10%. Type B2 and C lesions were encountered in 56% of the cases. More than one angiostent was implanted in 14 vessels and multiple stenting was accomplished with the use of different stents in 8 coronary arteries. No major complications were reported. The post-procedural minimal luminal diameter was 3.2 +/- 0.4 mm with a mean diameter stenosis of 1.4 +/- 3.7%. In 25 cases (49%) major side branches raised from the stented segment and in all but one remained patent. In conclusion, the implantation of the angiostent is safe, feasible and effective, as it can be easily deployed at the lesion site, used for the treatment of complex lesions and preserves the patency of jailed side branches. PMID- 9738331 TI - [Anticardiolipin antibodies and early infarct of the myocardium]. AB - The relationship between anticardiolipin antibodies (aCL) and acute myocardial infarction (AMI) is still controversial. The purpose of this study was to investigate the prevalence of aCL in young patients (age < or = 45 years) with AMI; record the aCL titre during different days of disease; assess the relationship between aCL titres and in-hospital myocardial infarction complications. The aCL were measured in 108 consecutive patients and in 31 controls (ELISA method). High aCL levels (IgG or IgM) were found in 19/108 (17.6%) patients and 5/31 (16.1%) controls (NS); aCL titres were similar in different days after AMI and did not differ in controls and in patients with or without early myocardial infarction complications. In conclusion, the aCL levels are not elevated in AMI patients, do not change during the early stage of the disease and are not associated with in-hospital complications. PMID- 9738332 TI - [Dispersion of junctional tissue and its consequences in a case of sudden infant death]. AB - This article reports a case of sudden infant death syndrome in a 1-month-old male. Some important abnormalities of the cardiac conduction system were found, consisting of persistent fetal dispersion of the atrioventricular node and His bundle, accessory atrioventricular pathways of Mahaim type and resorptive degeneration. These changes could be considered as a morphological substrate for cardiac arrhythmias. PMID- 9738333 TI - Successful ablation of atrioventricular nodal reentry tachycardia in a patient with persistent left superior vena cava. AB - We report the case of a patient with atrioventricular nodal reentry tachycardia associated with persistent left superior vena cava draining to a large coronary sinus. Successful ablation was performed at the level of the superior lip of the coronary sinus ostium. PMID- 9738334 TI - "Ventricularization" of the pulmonary artery pressure curve: a hemodynamic sign of proximal pulmonary embolism. PMID- 9738335 TI - [The endocrinologist]. PMID- 9738337 TI - [Introduction of intracytoplasmic sperm injection not rushed. Comment on M. Heisenberg: "Intracytoplasmic sperm injection and quality of the embryos"]. PMID- 9738336 TI - [Basic training in cardiology: theory and reality]. PMID- 9738338 TI - [Entero-hemorrhagic Escherichia coli--a new epidemic in central Europe?]. PMID- 9738339 TI - [Cryptosporidiosis and microsporidiosis in immune deficiency]. PMID- 9738340 TI - [A case from general practice. Children and the elderly especially at risk. Case report of a 76-year-old patient with EHEC infection]. PMID- 9738341 TI - [What blood pressure goal is sensible? Treatment of arterial hypertension- current results of the HOT Study (Hypertension Optimal Treatment Study)]. PMID- 9738342 TI - [Glycerol trinitrate and coronary heart disease. Value of prophylactic administration within the scope of exercise therapy]. PMID- 9738343 TI - [Disorders of porphyrin metabolism. 1: Pathophysiology , classification and clinical aspects]. PMID- 9738344 TI - [National health insurance refused high dose chemotherapy. "Death sentence" based on financial considerations?]. PMID- 9738345 TI - [Non-nucleoside reverse transcriptase inhibitors. AIDS long-term management]. PMID- 9738346 TI - [Do "health assistance needs" or "activities of daily living" determine the level of care? A multivariate analysis]. PMID- 9738347 TI - [Resources for home nursing care: willingness to undertake care by persons not involved in nursing services]. AB - In Germany the majority of people in need of nursing care are attended to at home. This is also promoted by the German nursing care insurance. Considering the increasing number of people in need of care more (people who care for them) are required. The article describes the readiness to care of a representative sample of 4806 people who are actually not involved in nursing care activities. The results show that the readiness to give nursing care is highly dependant on the care recipient. Independant from age, sex or social class, there is an strong readiness concerning care of the partner. Regarding other relatives (children, parents) or non-related persons like friends or neighbours, women show greater readiness than men to take over nursing care activities. Age and social class provoke differences, but not in such a homogenious way. All in all the readiness to provide nursing care is surprisingly high; it is to be hoped that it leads to realisation, in case of need. PMID- 9738348 TI - [Partial inpatient treatment as a responsibility in psychosomatic rehabilitation]. AB - Day clinic rehabilitation is part of the development of the medical rehabilitation system. Psychosomatic rehabilitation faces the task of developing a concept of day treatment and of collecting experiences with such models in clinical practice. The basis of this task consists in regulations laid down by the social security insurance authorities which serve as a frame for the development. The main points of these regulations are presented and their relevance for psychosomatic rehabilitation is outlined. The institutional and conceptual establishment of day treatment presents psychosomatic rehabilitation with a number of problems that are particular in this area and which are discussed in detail. Besides that, consequent elaboration of day treatment could stimulate further development of psychosomatic rehabilitation in general and help to overcome prevailing shortcomings of this part of the rehabilitation system. PMID- 9738349 TI - [Trends in average life expectancy in the Saxony Free State--results of a cause of death-specific table analysis]. AB - We conducted a specific table analysis on mortality causes that was mainly oriented on the lines of the chapters of ICD-9 to explore the average life expectancy for the population in Saxony particularly since 1988. The drop in average life expectancy of males observed since 1990 can be fully explained only for those around 18 years of age by "accidents" as the cause of death. Between 30 and 55 years of age there were additional losses of average life expectancy via the mortality causes "diseases of the digestive tract", "neoplasms" and "psychiatric diseases". The significance of neoplasms and of psychiatric diseases is receeding in respect of the average life expectancy. In 1991 there was an increase in the incidence of suicide in the age bracket around 51 years in males. Seen on an overall scale across all age brackets the development of the suicide rate has increased average life expectancy. Significant average life expectancy increase developed in the range of death causes due to embryonic or congenital damage as well as cardiovascular diseases, diseases of the respiratory system and infectious diseases. As far as the female population is concerned, only accidents and diseases of the digestive tract had a negative effect on average life expectancy in 1990 and 1991 whereas the negative effect exercised by the mortality cause "neoplasms" is systematically flattening out. There are excessive gains in life expectancy in respect of cardiovascular diseases, clearly in the range of embryonal and congenital diseases and remarkably in respect of suicide. How many of these specific differentiations of causes of death are due to changes in coding, has not yet been assessed. Leaving aside the unnatural causes of death these figures point to a marked influence of improved medical care on average life expectancy. PMID- 9738350 TI - [Quality circles in expert assessment as an instrument in quality management]. AB - An intercomparison programme was carried out as part of the quality management of medical assessments in the public health service in Baden-Wurttemberg. All physicians who performed medical assessments in January 1997 were invited to participate. They received a case description adopted from medical records of a precedent case (56-year-old head of a department in an administrative authority, severely disabled after left-sided stroke with residual right-sided partial hemiparesis, reduced work performance), and were asked to assess whether the patient was permanently unable to work. Access to medical expert opinions was requested. Participation was voluntary. Assessments were reviewed twice as to whether five prospectively defined evaluation criteria were fully met, or only partially or not at all. Participants and reviewers remained unknown to each other. Among 246 eligible physicians, 103 returned an assessment (43%). Forty five respondents had requested additional expert opinions. Anticipations for five criteria were met to their full extent with regard to: formal construction, all submissions; case description, 84%; sociomedical diagnosis, 65%; description of ability to work, 53%; and response to the question posed, 75% of submissions. The programme required considerable work time in its preparatory, field, and evaluation phases. The procedure was however found to be practicable, and a suitable element within a quality management programme can contribute towards motivation, quality assurance and the identification of possible deficits, and thus indicate possible topics and issues for continuing education and quality control programmes. PMID- 9738351 TI - [Unconventional diagnostic and therapeutic methods in environmental medicine]. AB - In the sphere of environmental medicine--analogous to other fields like oncology and chronic diseases--not only proven and approved methods, but also unconvential methods are offered, without evidence of efficacy. The application of these methods has the possible consequence of wrong diagnosis and malpractice. Examples are discussed such as Kirlian photography, electroacupuncture according to Voll, bioresonance diagnosis/therapy, kinesiology, regulation therapy according to Rost, "clinical ecology" according to Runow with, among others, the provocation/neutralisation test, a vaccination therapy with E. coli and finally electrosmog as an environmental noxa. Concerning the admissibility of contested methods, statements of medical specialist societies, judgements, and the law of medical products are quoted. In conclusion, the question of the origin of the ideas and alleged results of unconvential medicine is followed up and conclusions are drawn. PMID- 9738352 TI - [Community health reporting exemplified by the city of Nurnberg]. AB - Taking the example of Nuremberg, a city of half a million inhabitants, the article illustrates how, despite very restricted resources, the local public health department was able to establish a series of health reports dealing with different topics. Treatment of a number of topics was only possible through cooperation with the local university. Initially two health reports were produced on a regional basis for the purpose of supporting health promotion in particular areas of the city. Whereas the first report attempts to give a comprehensive picture of health relevant living conditions in a certain area, the second report concentrates on an analysis of the situation of children and young people in a different part of the city. Another area of concern deals with the gathering of specific health related data. The data from a pediatric health sentinel in Nuremberg were combined with data on climate and air pollution in the years 1995 and 1996. Various dissertation topics were useful in the analysis of everyday public health data. Most of these results have been published in a separate reader. They include an analysis of the reports regarding pregnancy counciling, standardised reports of the HIV antibody tests as well as an analysis of the causes of infant mortality based on death certificates. Finally, a comparative study of basic health data from various cities is in progress. It is hoped that this will offer a basis for an evaluation of the health situation in Nuremberg. PMID- 9738353 TI - [Public health relevant levels of pollutants in soil. Considerations for preventive, environmental health protection]. AB - A working document on nation-wide applicable health-related guiding levels of environmental pollutants in soil has been prepared. These levels are intended to be the base for the administrative regulation of soil contaminations in connection with the German "Bundesbodenschutzgesetz" (Federal law on the protection of soil). Legislation demands these base levels to be intensely related to protection against serious health effects (i.e. health hazards). Looking more closely at the toxicological deduction of these values and at some values extracted from that process some uncertainties become obvious with regard to a clearcut limit between dangerous and noxious levels in soil. Indeed there are some arguments which suggest a more conservative approach. Health-care based recommendations should be introduced also at lower contaminant levels in soil. Therefore, development of an internal administrative manual on basic noxious values of soil pollutants (e.g. values below the threshold of danger) is suggested which may be useful for health authorities to cope with special problems in individual cases. PMID- 9738354 TI - [Dichlorvos insect strips indoors: pollution and risk assessment]. AB - Exposure to Danger and Assessing the Hazards: Application of the highly potent pest control agent dichlorvos (DDVP) involves the hazard of effects on the central nervous system and a risk of human cancer. Consumers use dichlorvos in pest strips to protect their rooms from insects. This undifferentiated pest control can lead to chronic exposure. Over several weeks we found an indoor air concentration of dichlorvos between 60 and 1300 micrograms/m3 in 34 m3 experimental-room (chamber). Textiles and foodstuffs in this room were also contaminated. Beside the inhalative intake the oral and dermal intake can also lead to an additional loading of the residents. The ADI-resp. DTA-value of 4 micrograms/kg body-weight and day is exceeded about 10-15 times on the average by inhalative exposure only. PMID- 9738355 TI - [Current hepatitis B epidemiology in Germany]. AB - Hepatitis B (HB) is the most common occupational hazard for health care workers. On the other hand 90% of the notified HB cases in Germany are observed among the general population (including other high-risk groups like i.v. drug abusers etc.). About 350 million persons all over the world are carriers of the hepatitis B virus (HBV). The availability of effective HB vaccines since 1982 made a control of HB possible. In this communication we report on epidemiological changes due to hepatitis B vaccination in a large university hospital in Germany. Health care workers and non-health care workers occupied at Freiburg university hospital were tested for HBV markers in 1984/85 (n = 4218), 1989/90 (n = 4081) and 1994/95 (n = 4022) during routine occupational health check-ups. Vaccinations were performed using a plasma-derived vaccine (1984/85) or a vaccine obtained by genetic engineering, respectively (1989/90, 1994/95). In 1984/85 prevalence of anti-HBs/HBc in German health care workers (12.4%) was 2.5 times higher than the one in non-health care workers (4.9%), in 1994/95 anti-HBs/HBc prevalence in both groups (4.4 vs. 4.5%) was comparable. On the other hand HBV carriage in persons occupied in professions without blood contact increased from 1984/85 (0.5%) to 1994/95 (1.1%). Therefore, the number of HBV carriers (ca. 1.1 millions) in Germany can be roughly estimated. Our data indicate a high degree of effectiveness of hepatitis B vaccines. Vaccination programme for the general population in accordance with German (STIKO) and WHO recommendations are necessary for the control of HBV infection. PMID- 9738356 TI - [Common goal determination in occupational health--experiences in North-Rhine Westphalia with a system of health conferences]. PMID- 9738357 TI - [Hyperreactivity in Mendel-Mantoux tuberculin test]. PMID- 9738358 TI - [Etiopathogenic, clinical-diagnostic and therapeutic aspects of the burning mouth syndrome. Research and treatment protocols in a patient group]. AB - BACKGROUND: Burning mouth syndrome (BMS) is a frequently seen pathology characterised by burning tongue and oral pain without macroscopic structural lesions to the mucose. BMS etiopathology isn't known and therapy is merely empirical and unsatisfactory. METHODS: To evaluate the hypothesis that this syndrome would originate by a small diameter peripheral neuropathy combined to a mucosal trophic lesion, 37 patients, (7 male, 30 female, between 36 and 79 years, mean 54 years) affected by BMS, consecutively observed in our dispensary were submitted to a series of examinations and to therapeutical approach used in neuropathic painful syndromes. All patients were submitted to a complete stomatological exam and X-ray pantomography to exclude mucosal macroscopical lesions and dentistry illnesses. All patients executed sierological exams (glycemia, etc.), neurological exam, tongue and foot dorsum quantitative sensory examination, tongue and face telethermography. A few patients (3 male, 10 female; age 34 to 53, mean 49) were submitted to mucosal tongue biopsy, analyzed by optic microscopy and immunofluorescency following treatment with anticytoplasmatic neuronal proteins antibodies (protein gene product 9.5). RESULTS: These examinations showed subclinical polyneuropathy in 50% of patients. In particular, a loss of function in small diameter nervous fibres in about 50% of patients was observed. Histological examination of tongue mucose revealed a moderate atrophy in 70% patients. CONCLUSIONS: All patients were submitted to an antalgic therapy, with non-antiflammatory drugs used in neuropathic painful syndromes (quercetine, antiepileptic drugs benzodyazepinein and gabaergic, topical application of capsaicine solutions). PMID- 9738359 TI - [Clinical anatomy of the human mental foramen]. AB - BACKGROUND: The great diffusion of the surgical technics in oral implantology and the progress of the radiological imaging produces some interest for the clinical anatomy of the mental foramen (MF). The study, in addition to the measurements that define it, considers others anatomical features of practical utility and the variableness of them. METHODS: In the Anatomic Institute of the Bologna University, it has been made a morphometric revision of the MF on 100 dried mandibles of normal young adults (78 males, 32 females) random chosen with the complete integrity of the dental apparatus and of the mandibular bone criteria. The measurements have been made by anthropometric methods on the two sides of the same mandible (n = 200) and for everyone has been reported in the tables the medium, the maximum and the minimum values with their specific variation interval. The results are applicable to the common work conditions because they take in consideration among the specific characteristics of the MF in addition to the seat, the course of the mandibular canal, the thickness and the height of the mandibular bone. RESULTS: To the clinical and diagnostic imaging object the medium values of the anatomical measurements can be considered sufficent. To anthropometric, anesthesiologic and surgical aims it is also necessary the knowledge of the maximum and the minimum values and the variation interval. In the living man the anatomicomedical study of the MF is made by the diagnostic imaging, especially by computed tomography with specific algorithm, because it makes possible absolutely exact measurements. CONCLUSIONS: Finally, the specifications on the MF provided by the present study are important not only for an anatomical but also for a practical point of view because they are a datum point value in the patient clinical management. PMID- 9738360 TI - [Pharmacologic therapy of cranio-cervico-mandibular disorders. Review of the literature]. AB - The recommended treatments of craniomandibular disorders (CMD) are drug therapy, physiotherapy, relaxation procedures, occlusal therapy with and without splint. The purpose of this study is to carry out a survey of the literature on drug therapy in patients affected by CMD. It is essential to recognize the cause of pain (muscular or articular) and the phase of disorder (acute or chronic) in order to establish an adequate pharmacological protocol for each type of CMD. Non steroidal anti-inflammatory drugs (NSAID) are generally accepted for treatment of internal derangement and myofacial pain, sometimes in association with benzodiazepine. PMID- 9738361 TI - [Necrotizing sialometaplasia of the submandibular gland. Report of a case]. AB - A case report of swelling of the right submandibular gland, with spontaneous remission, is described. Clinical, ultrasonographical and cytological features suggest that the swelling could be ascribed to a necrotizing sialometaplasia. The etiopathological hypothesis and pathology of the lesion are presented, and the differential diagnosis with other spreading lesions of the submandibular gland is discussed. PMID- 9738362 TI - [A case of a midline cervical cyst]. AB - Midline cervical cysts are one of the most common embryological anomalies of the neck. They are due to the failure of a complete obliteration of the thyroglossal duct. This endodermal structure arises from the floor of the mouth and proceeds caudally in the midline of the neck until it reaches its final position around the trachea. Finally, the inferior portion of the thyroglossal duct develops into the median lobe of the thyroid gland whereas its cranial portion disappears. In the present paper, the clinical feature and the surgical treatment of a case is described. PMID- 9738363 TI - [Hereditary hemorrhagic telangiectasia (Osler-Rendu-Weber disease) Management of epistaxis and oral hemorrhage by Nd-Yag laser]. AB - BACKGROUND AND AIMS: The paper reports the use of Nd-Yag laser treatment for some major otolaryngoiatric symptoms in hereditary hemorrhagic teleangectasia (Rendu Osler-Weber disease), focusing on the importance of epistaxis and oral hemorrhage as a cause of continuous hemoreintegrative treatment, without forgetting the constant psychophysical stress undergone by these patients. MATERIALS AND METHODS: Photocoagulation of teleangectasia in the ENT district has been used by the authors since January 1994. A total of 8 patients, 5 males and 3 females, have been treated aged between 31 and 65 years old. During the first year specialist outpatient controls were performed every two months, after which the follow-up became four-monthly. RESULTS: The authors provide a detailed illustration of the methods used, the choice of power, and the advantages and results obtained, emphasising the outpatient nature of treatment, excellent patient tolerability and the low cost to the health service. CONCLUSIONS: In the light of these results, the Authors underline the unquestionable usefulness of Nd Yag laser in the treatment of a few of the undoubtedly more stressing symptoms of Rendu-Osler-Weber disease. PMID- 9738364 TI - [Management of postoperative pain in stomatology with ibuprofen L-arginine and naproxen]. AB - BACKGROUND: The aim of this prospective, randomized study is to compare the efficacy and safety of ibuprofen L-arginine and naproxen in the treatment of postoperative dental pain. METHODS: Seventy patients undergoing removal of impacted third molars were randomly allocated to receive 4 hours after surgery a single oral dose of either ibuprofen L-arginine 400 mg or naproxen 550 mg. Ten patients dropped out from the study because they took the study drug before the allowed time. Using a self-rating record, patients rated their pain and its relief for 1 hour after the drug administration. Remedication, if needed, and mean time of remedication were also recorded. RESULTS: A statistically significant reduction in pain scores with respect to the baseline values was recorded 5 minutes and 15 minutes after the drug administration in the ibuprofen L-arginine and in the naproxen-treated group, respectively. The summed pain intensity difference (SPID) over 60 minutes resulted significantly higher in the ibuprofen L-arginine than in the naproxen-treated group. A complete abolition of pain 60 minutes after medication was obtained in 12/28 patients (42.9%) in the ibuprofen L-arginine and in 5/32 patients (15.6%) in the naproxen-treated group, respectively (p = 0.04). Number of patients requiring remedication, mean time of remedication and drug related adverse effects did not significantly differ in the two treatment group. CONCLUSIONS: Global evaluation of the drugs by the patients showed ibuprofen more effective drug than naproxen. PMID- 9738365 TI - [Management of hematologic systemic diseases and rare tumor entities with manifestations in the oromaxillofacial area]. AB - Malignant lymphomas, hematological malignancies, sarcomas, occult head and neck primaries, Merkel cell carcinomas and malignant melanomas are among the tumors that are rarely seen in the head and neck region. Almost 20% of patients with acute leukemia initially present with symptoms of the oral cavity (ulcerations, gingival hypertrophy, etc.). If a malignant lymphoma is suspected, the lymph node should be removed in toto to ascertain diagnosis. Furthermore, in order to make sure that the immunohistological work-up or electron microscopic analysis is adequate, the pathologist should be informed prior to extirpation of the suspicious lymph node. The same diagnostic procedure is indicated for metastases from undifferentiated or small cell cancer of an unknown primary. Metastatic squamous cell or undifferentiated carcinoma to a solitary cervical lymph node from an unknown primary can be cured by multimodal therapy (extirpation, radical neck dissection and adjuvant radiation) in 30% of the cases. Following polychemotherapy, long-term survival may also be achieved in disseminated stages of undifferentiated carcinoma or poorly differentiated adenocarcinoma in an occult primary. When osteosarcoma of the jaw is suspected, core biopsy has to be planned carefully: in order to prevent tumor seeding, the needle track has to be excised during definitive surgery. Most authors propose (neo-)adjuvant chemotherapy (or chemoradiation) for head and neck osteosarcoma, especially when additional risk factors, i.e. a large primary or poorly differentiated sarcoma, are present. Patients with positive margins should receive adjuvant radiotherapy in soft tissue sarcoma. Local control of angiosarcoma is possible exclusively by radiation. Adjuvant radiation is also indicated in Merkel cell carcinoma. Because this tumor spreads in a "cascade" fashion, elective node dissection may also provide a chance for cure. Excision with wide margins is the principal therapeutic step in malignant melanoma. Due to the anatomic localization, adequate resection may not be possible in mucosal melanoma of the head and neck. When regional lymph nodes are involved, radical lymph node dissection and adjuvant radiation have to be added to the therapeutic concept. There is an emerging role for adjuvant interferon alpha in intermediate and high-risk melanoma. PMID- 9738366 TI - [Value of (F18)-2-fluorodeoxyglucose PET scanning in staging mouth cavity carcinoma. Comparative evaluation of PET findings before and after preoperative radiochemotherapy with histological and computerized tomography findings]. AB - OBJECTIVE: The lack of sensitivity and specificity of conventional imaging techniques based on morphological critera is responsible for considerable limitations in the staging and surveillance of oral cancer. Therefore, this study investigates the contribution of [F18]-2-fluordesoxyglucose (FDG) positron emission tomography (PET) to tumor management with special regard to lymphnode involvement and therapeutic monitoring after radiotherapy. DESIGN: Prospective observational study. PATIENTS: Twenty-one patients with advanced oral cancer, predominantly T3/T4. INTERVENTION: FDG-PET scans before and after preoperative radio(chemo)therapy. Standardized uptake values (SUV) were determined for the tumor site and lymphnode areas. PET scans were correlated to histological findings after ablative tumor surgery. RESULTS: FDG-PET yielded superior sensitivity and specificity for tumor and lymphnode assessment. The effect of radiotherapy was reflected by the metabolic activity of the tumor, which shows a close correlation between the decrease of FDG uptake and histologic tumor regression. PET detected distant metastases and simultaneous tumors. CONCLUSION: FDG-PET is a challenging imaging technique with the potential to improve staging procedures for oral cancer. In the monitoring of metabolic activity of the tumor in the course of radio(chemo)therapy, FDG-PET allows objective measurement of the treatment response. PMID- 9738367 TI - [Low-dose computerized tomography of the jaw bone in pre-implantation diagnosis. Limits of dose reduction and accuracy of distance measurements]. AB - Absorbed radiation doses delivered by computed tomography and panoramic radiography were measured in 16 anatomic sites using a head and neck phantom and thermoluminescent dosimetry. The recommended kilovoltage and scan time for dental scanning was reduced step by step, rating the quality of the low-dose scans. A reduction of up to 76% could be achieved without loss of diagnostic accuracy. Measured absorbed radiation dose ranges from 0.30 mGy (thyroid) to 29 mGy (skin) at 187.5 mAs and 1.0 mm-slices (25 mm scanning distance for maxilla, 30 mm for mandible). After reduction to 45 mAs, 0.07 mGy (thyroid) to 6.9 mGy (skin) was measured. Distance measurements on human jaw specimens were compared with corresponding CT image measurements. Average deviation was 0.1-0.3 mm. A dose reduction of 75% had no effect on the results. However, the doses of CT-scans reduced by 76% exceed by an average factor of 10 the doses of conventional panoramic radiography. Therefore, CT should be reserved for the planning of complex implant treatment in the direct vicinity of the maxillar sinus and nerves and for multiple implant insertion. PMID- 9738368 TI - [Perioperative antibiotic prophylaxis in orthodontic bone operations of the facial skull]. AB - Intraoral orthognathic surgical procedures are clean-contaminated operations because of the facultative pathogenic flora of the oral cavity. Without antibiotic prophylaxis in this kind of operation a postoperative wound infection can be expected in 20-31% of cases. Therefore, a retrospective analysis of the clinical course of 545 patients with various dentofacial deformities was performed to evaluate the importance of perioperative antibiotic prophylaxis. The total rate of wound infections was 2.8% and lower than rates published in other comparable studies without antibiotic prophylaxis. The rate of wound infection after single mandibular osteotomy (4.1%) was significantly higher than the analogous amount after single maxillary osteotomy (0.8%) because of the mechanical strain of the surgical wound in the region of the mandibular angle and because of the high retention of food particles and other decay products in this area. The recognized rise in the wound infection rate by increasing the length of operation demonstrates that the extent of bacterial contamination of the surgical wound depends on the dimension and duration of surgery. Bacterial flora caused the noticed wound infections in 61.5% of cases aerobic-anaerobic mixed infections, which is in accordance with the results of similar publications. With increasing length of antibiotic prophylaxis, an increase in the incidence of antibiotic-associated side effects and in the wound infection rate could be observed. Therefore, an antibiotic prophylaxis which exceeds 3 days does not seem to be useful. In this study, in which cephalosporins were mostly used, the total rate of antibiotic-associated side effects (4.6%) was lower than analogous amounts of other groups of antibiotics. The analysis demonstrates that a short term prophylaxis with broad-spectrum cephalosporins such as cefatoxime of 48 h is very effective and useful. PMID- 9738370 TI - [Dissection of the internal carotid artery with ascending thrombosis of the ophthalmic artery as a rare differential diagnosis of post-traumatic amaurosis]. AB - The case of a 30-year-old patient is reported who presented with a right-sided amaurosis fugax and a simultaneous frontobasal fracture that had occurred after a fall. By CT angiography and MRI angiography obstruction of the ophthalmic artery following a dissection of the internal carotid artery with ascending thrombosis could be demonstrated. There was a lack of further neurological deficits because of sufficient perfusion of the cerebral arteries by the arterial circle of the cerebrum. Among the etiological variety of types of posttraumatic amaurosis, this rare cause should be taken into consideration in the etiology, diagnosis and therapy, if appropriate. PMID- 9738369 TI - [Results of transcranial and subcranial management of fractures of the naso ethmoid-orbital system in complex midfacial fractures]. AB - Injuries of the nasoethmoid-orbital (NEO) region are associated with midfacial fractures or fractures of the frontobase in over 90% of all cases. In up to 70% fractures of the skull base run through the roof of the ethmoidal bone or the lamina cribrosa. There are different surgical approaches for the treatment of these complex fractures. Between 1990 and 1997 50 patients with midfacial fractures in association with NEO fractures were treated in the Klinik fur Mund-, Kiefer- und Gesichtschirurgie, Kantonsspital Luzern, Switzerland. Of these, 25 had suffered midfacial fractures combined with fractures of the nasoethmoid orbital and frontobasal region and were treated via a transcranial approach. The other 25 patients with midfacial and NEO fractures without involvement of the frontobasis were managed by subcranial incisions. A total of 47 patients were followed up for up to 4 years. The results were reevaluated retrospectively. There was no case of secondary liquorrhea, intracranial or ethmoidal infection. Our therapeutic concept of transcranial and subcranial management of NEO fractures in combination with frontobasal and midfacial fractures is demonstrated. PMID- 9738371 TI - [Single-cystic ameloblastoma at an early age. 2 case reports]. AB - Two cases of unicystic ameloblastoma in the mandible of young children are described. Conservative surgery is justified in children due to the lower risk of recurrence compared to cases of solid ameloblastoma and the risk of growth disturbance after radical surgery. However, long-term follow-up is necessary. PMID- 9738372 TI - [Gigantic proliferating trichilemmal cyst of the scalp with central carcinoma and lymph node metastasis]. AB - We report the case of a 77 year old woman with a giant proliferating trichilemmal cyst of the scalp and a central squamous-cell carcinoma. Six months after radical tumor-excision and dissection of the locoregionary lymph nodes the patient developed retroclavicular lymph node metastases. The tumor develops from cells of the hair matrix and tends to recur after excision; in rare cases malignant transformation may occur. We suggest radical excision and complete histological examination of the tumor. In cases of malignancy, lymph node-dissection is sometimes necessary. In these cases close postoperative follow-up of the patient is mandatory. PMID- 9738373 TI - [Gustatory rhinorrhea after hemiresection of the maxilla]. AB - A case of gustatory rhinorrhea after maxillary resection is reported about. The symptoms can be explained as defective regeneration of secretory nerves, especially of the nasopalatinal nerve. PMID- 9738374 TI - [Choroid neovascularization in senile macular degeneration. 1 hear follow-up after radiotherapy]. AB - Choroidal neovascularization (CNV) associated with age-related macular degeneration is the major cause of legal blindness in Europe and the USA in patients aged more than 65 years, but Chakravarthy et al. has reported that radiotherapy has a beneficial effect on visual acuity. METHODS: Since March 1996 we have treated 56 patients in cooperation with the Department of Radiotherapy at the Technical University in Munich. The total dose with external beam radiotherapy was 16 Gy in 8 fractions, delivered through an anterior oblique axis to spare the lens. Before the treatment and 3, 6 and 12 months after therapy, we performed a standardized visual acuity and contrast-sensitivity test (ETDRS, Pelli Robson Chard) and fluorescin angiography 6 and 12 months after therapy. RESULTS: Twenty-five angiograms showed well-defined CNV and 31 not well-defined CNV. Six months after the treatment 15 patients had stable visual acuity within one line. Twenty-seven patients had lost more than one line of visual acuity. There was no difference between well and not well defined CNV's. One year after treatment the visual acuity remained stable within one line in 4 patients, no patient had an increase of two lines or more and 17 patients lost more then 2 lines of vision. We saw no side effects other than sicca symptoms in 3 patients. CONCLUSION: In our opinion, these results do not show that radiation treatment has a real beneficial effect on visual acuity. Further randomized studies are needed to demonstrate the efficiency of this treatment for choroidal neovascularization in AMD. PMID- 9738375 TI - [Low dosage, fractionated, percutaneous teletherapy of subfoveal choroid neovascularization]. AB - AIM: The effect of low-dose fractionated percutaneous teletherapy on visual acuity and the changes in subfoveal neovascular membranes (SNM) in age-related macular degeneration (ARMD) were studied. PATIENTS AND METHODS: Forty-four patients (aged from 56 to 86 years) were treated. Best distant and near visual acuity was assessed before (initial visual acuity; IVA) and 5 weeks, 3, 6 and 12 months after teletherapy. Fluorescein angiography was done before and 3 and 12 months after radiation. Patients were divided into different groups by IVA for analysis. Mean follow-up was 9 months. Teletherapy was done by a 9 MeV linear accelerator through a lateral port in the half-beam technique with a single dose of 2 Gy to a total dose of 20 Gy within 20 days. RESULTS: No severe negative side effects have been observed. Four patients reported epiphora. Visual acuity decreased one line in the group with an IVA of 0.05-0.2. The group with IVA 0.3 0.5 remained unchanged during the observation period. We found a tendency for increased visual acuity in group IVA > or = 0.6. CONCLUSIONS: Teletherapy seems to have an influence on visual acuity, SNM, and metamorphopsia. IVA and duration of thillness play important roles. We cannot say if there is a persistent effect, as these are preliminary results. The longest follow-up so far has been 12 months. In order to better evaluate the potential of radiotherapy, this study must be pursued and coupled with further studies analyzing the effect on SNM in ARMD. PMID- 9738376 TI - [Acute retinal necrosis syndrome. Argon laser coagulation for prevention of rhegmatogenic retinal detachment]. AB - BACKGROUND: ARN syndrome follows severeintraocular infection by herpes viruses and primarily affects the peripheral retina. Following scar formation, despite antiviral treatment, rhegmatogenous retinal detachment occurs very often. Prophylactic argon laser photocoagulation has therefore been proposed. We report our experience. PATIENTS: We treated five patients presenting clinically with advanced unilateral ARN with acyclovir. All eyes received a prophylactic confluent double row of argon laser treatment (500 microns, 0.2 s, gray-white lesions) central to the affected area as soon as was possible, depending on the vitreous clouding. Four patients were treated with Aspirin. RESULTS: One of the five patients had a peripheral rhegmatogenous retinal detachment that was limited by the argon laser row. Another patient had a tractional detachment needing vitreoretinal surgery. Two eyes developed vitreal hemorrhage of unknown origin. CONCLUSION: A lower rate of rhegmatogenous retinal detachments than expected occurred post-laser treatment. Vitreal hemorrhage was more frequent than previously reported. The bleeding probably originated from anterior retinal neovascularization and may have been enhanced by Aspirin treatment. We recommend early prophylactic argon laser photocoagulation in all ARN patients in agreement with the results of previous studies. PMID- 9738377 TI - [Experiences with "reversed tip and snip" phacoemulsification]. AB - The endothelial cell count after phacoemulsification serves as a sensitive indicator for the level of corneal traumatization caused by different phacoemulsification techniques concerning the used phacotime and phacoenergy/s. PATIENTS: In a prospective and randomized study, the "reversed tip and snip" technique and the "divide and conquer" technique were performed in groups of 30 patients each. The corneal endothelial cell count was measured preoperatively as well as 4 weeks and 3 months postoperatively. In a second study the phacotime and phacoenergy used/second and the endothelial cell loss were compared between patients 1-30 and 31-60 who were operated on with the "reversed tip and snip" technique, demonstrating the learning curve of this technique. RESULTS: The endothelial cell count showed significant (P < 0.001) reduction by about 10% after the "reversed tip and snip" technique and by about 15% after the "divide and conquer" technique. The latter produced significantly greater cell loss. The phacotime (mean: 22-17) and phacoenergy/s (mean: 23-16) were reduced significantly (P < 0.05) when patients 1-30 were compared with 31-60. CONCLUSION: The "reversed tip and snip" phacoemulsification technique produces less endothelial cell loss than the "divide and conquer" technique. There is a short learning curve by using the "reversed tip and snip" technique. PMID- 9738378 TI - [Frequency of Nd:YAG capsulotomy after implantation of PMMA and silicon intraocular lenses]. AB - The aim of this investigation was to ascertain the frequency of secondary cataract after implantation of PMMA or Silicone IOLs. PATIENTS AND METHODS: In our series of patients we implanted 900 PMMA IOLs in a period of 4 years. In the following 3 years we implanted 1600 silicone IOLs. After the operation, a Nd:YAG laser capsulotomy was indicated there was a reduction in visual acuity and the patient asked for improvement. With the help of computer documentation we had the possibility to follow up a period of 6 years after PMMA and 3 years after silicone IOL implantation. RESULTS: (1) PMMA IOL: After implantation of PMMA IOLs we found a frequency of 22% capsulotomies during the investigation period of 6 years; 4.9% of them occurred during the 1st year and 8% during the 2nd year. (2) Silicone IOL: During the control period of 3 years we found a capsulotomy rate of 9.9%. Most happened in the 1st year. CONCLUSIONS: Our 22.9% capsulotomy rate after PMMA IOL implantation is lower than the rate mentioned in the literature. We noticed a peak of capsulotomies in the 2nd year after the operation. During a control period of only 3 years after implantation of silicone IOLs we found a capsulotomy rate of 9.9%. PMID- 9738379 TI - [Toxocara canis infection. Environmental parasitologic and epidemiologic studies]. AB - Due to the large number of differential diagnostic possibilities, the etiology of endogenous uveitis is still hard to determine. One reason for uveitis may be the occurrence of parasites. However, too little attention is paid to this underlying disease. METHODS: To identify certain sources of infection, ovoscopic probes of 98 dog feces from the urban area of Halle were taken. The material was collected from children's playgrounds, parks and swimming pools. A retrospective analysis of patient case data from the records of the University Eye Hospital for the years 1986-1995 complemented the environmental and parasitological examination of patients with Toxocara canis and T. cati infections. RESULTS: From the ovoscopic examinations, 16% of the fecal samples contained T. canis eggs. Seventeen percent contained coccicidal sporocysts. Eggs of Trichuris vulpis and not clearly identifiable nematodal larvae were found in 1% of the samples. The reasons for the morphology was, in one instance, suspected Echinococcus eggs. The fact that 12 or 15 parasitologically positive dog excrement samples were taken from the immediate vicinity of children's playgrounds and another three directly from these grounds should be regarded as particularly critical. The investigated patient case data records revealed for the period studied nine uveitic patients with T. conis infection. The diagnosis was based on the larva precipitation test in all cases. After correct diagnosis and appropriate treatment, the prognosis was favorable. The specific treatment had to be repeated in two patients. CONCLUSIONS: The environmental parasitological study presented proves the relatively high prevalence of pathogenic causal agents in dog excrement samples from the area of urban recreation and leisure grounds, in particular, children's playgrounds. Parasite infestation is a mirror of the efficiency of hygienic and social measures. Public health education must be reinforced. Differential diagnosis of chronic endogenous eye diseases must pay more attention to infections by parasites. PMID- 9738380 TI - [Opening of lacrimal duct stenoses with endoscope and laser]. AB - BACKGROUND: Since the development of lacrimal endoscopes with which the mucosa and disorders of the lacrimal drainage system can be examined directly, the choice of surgical technique has changed decisively. Flexible miniendoscopes can be combined with a laser for the first time, allowing for exact localization of the stenosis and subsequent therapy. PATIENTS AND METHODS: Between January 1996 and February 1997, 12 patients with acquired lacrimal stenosis were treated with an Hyb:erbium-YAG laser. After endoscopic localization, the stenosis or stricture was opened using the laser. Bicanicular intubation into the nose was performed and was left for at least 3 months. Five patients were treated for stenosis of the lower canaliculus and seven patients for postsaccal stenosis. RESULTS: The longest follow-up period was 22 months and the shortest 9 months. Treatment was successful in 4 cases with a canalicular stenosis, but restenosis occurred in all cases of postaccal stenosis and in 1 case of canalicular stenosis, and required a DCR. CONCLUSIONS: At this time we conclude that disorders of the canaliculi can be successfully treated with endoscopic localization, opening with a laser and subsequent silicone tube intubation. A high rate of restenosis may be expected in cases of postsaccal stenosis. PMID- 9738381 TI - [LASIK for myopia correction. 2-year follow-up]. AB - BACKGROUND: Long-term results on LASIK are not available to date. We therefore evaluated the predictability, stability and complication rate after LASIK in moderate--to-high myopia. PATIENTS AND METHODS: We treated 70 eyes (41 patients) using the Automatic Corneal Shaper and the Keracor 116 excimer laser. Patients were followed for 1, 6, 12 and 24 months. Spectacle refraction, visual acuity, rate of retreatment, and patient satisfaction were evaluated. RESULTS: At 24 months the results were as follows: Myopia -5 to -9.9 D (n = 18): 94% within 1 D; regression between 1 and 12 (12 and 24) months > 1 D in 6% (6%); uncorrected acuity 20/40 or better in 83%; no loss of 2 ore more lines of visual acuity; 89% highly satisfied. Myopia -10 to -14.9 D (n = 12): 88% within 1 D; regression between 1 and 12 (12 and 24) months > 1 D in 20% (0%); uncorrected acuity 20/40 or better in 72%; 4% lost 2 or more lines of visual acuity; 96% highly satisfied. Myopia -15 to -29 D (n = 22): 33% within 1 D; regression between 1 and 12 (12 and 24) months > 1 D in 41% (18%); uncorrected acuity 20/40 or better in 7%; no loss of 2 or more lines of visual acuity; 67% highly satisfied. CONCLUSION: LASIK is an accurate, effective and stable procedure for correcting myopia of -5 to -10 D. Results are less precise in myopia up to -15 D, and some visual loss occurs in a number of patients. In myopia > -15 D, results are not satisfactory because of poor accuracy and low stability. PMID- 9738382 TI - [Intraocular pressure regulation after combined glaucoma and cataract operation]. AB - BACKGROUND: The success rate of combined glaucoma and small incision cataract surgery is not yet established. PATIENTS AND METHODS: Therefore, 56 eyes of 52 patients (mean age 79.0 years) having trabeculectomy combined with phacoemul sification and implantation of a silicone posterior chamber lens were retrospectively investigated. RESULTS: The mean follow-up was 21.6 weeks. Due to the combined surgery the mean intraocular pressure decreased significantly (p < 0.001) from 21.8 to 14.8 mmHg. Seventy-five percent of the patients did not need any antiglaucomatous medication, while 25% still had to use a local medication, but less frequently. The overall visual acuity increased significantly (p < 0.001) from a preoperative value of 0.24 to 0.52 postoperatively. CONCLUSIONS: The results of combined glaucoma and cataract surgery seem as good as those reported for two separate glaucoma and cataract procedures. PMID- 9738383 TI - [Value of anterior chamber lenses in developing countries. Results of a clinical study]. AB - There are estimated to be 20 million people blinded by cataracts, 80-90% of whom live in rural areas of developing countries where expert surgical resources are scarce. The majority of all cataract operations are still intracapsular extractions (ICCE). Aphakic correction using spectacles is problematical in developing countries. This study was undertaken to evaluate the safety of multiflex open loop anterior chamber intraocular lenses (AC IOLs). METHODS: A total of 2000 people attending Lahan Eye Hospital, South-east Nepal, with bilateral cataract were randomly allocated to receive in their first eye either ICCE with AC IOL (AC IOL group) or ICCE with aphakic correction (control group). All operations were performed by two ophthalmologists using a standardized technology and 4.5 x operating loupe magnification. Functional and best corrected vision was recorded. The primary outcome measure was poor vision after surgery, which was defined as a visual acuity of less than 6/60 at 1 year follow-up (WHO definition for severe visual impairment and blindness). FINDINGS: The median time needed to perform ICCE was 4.1 min and to perform ICCE with AC IOL 6 min. Of all study patients 91% were examined after 1 year. Five percent of the AC IOL group and 5.4% of the control group had a functional visual acuity of less than 6/60. Causes of reduced vision in the AC IOL group versus the control group were: correctable refractive error (22 vs 29), uveitis/secondary glaucoma (13 vs 2), endophthalmitis (4 vs 7), pre-existing eye diseases (4 vs 5), retinal detachment (0 vs 4), and corneal decompensation (0 vs 1). Of the control group, 24 patients were found to be functionally blind in the operated eye (vision < 3/60) because they did not wear their aphakic spectacles. Normal vision (WHO definition: > or = 6/18) was achieved best corrected in 89.9% of the AC IOL group and 93.2% of the control group. Analysis of additional long-term follow-ups (2-5 years post operatively) has not yet been completed. INTERPRETATION: This study provides evidence that in developing countries well-manufactured multiflex open loop AC IOLs can be implanted safely by experienced ophthalmologists after routine ICCE, avoiding the disadvantages of aphakic spectacle correction. PMID- 9738384 TI - [An ophthalmologic image databank on the Internet]. AB - The Internet has become a powerful, international source for information, and it has shown an exponential growth because of the ease of access and the immediate availability of information. We introduced a new ophthalmological atlas including ICD coding on the World Wide Web. So far, more than 500 lantern slides of typical and interesting ophthalmological findings have been selected and digitized. These pictures were integrated in a data base, which was arranged for ease and speed of search and retrieval. In its background, the data base contains a list of over 4700 ICD-encoded diagnoses to which the picture-documented findings are linked. Comments on pictures can be added by the author or by users. The data base contains several lists, such as a list of ICD codes and diagnoses, a list of all pictures with corresponding diagnoses, a list of all diagnoses and number of pictures, a list of those diagnoses for which the corresponding picture is available, as well as a list of comments on each picture. Special program scripts handle the user's search key words for diagnoses and extract the required information out of the data base, using Windows NT. Search results are presented on an automatically built-up webpage. To provide fast speed of search all pictures initially are shown in a small format (thumbnails) with little amount of data. The related full-size picture is retrieved by a single mouse click. Moreover, the name and institution of the author, diagnostical hints and comments on pictures by the author or by users are offered for each diagnosis available. The Giessen Ophthalmological Picture Atlas can be reached under www.med.unigiessen.de in the Internet. It allows a fast search free of charge from all over the world and, therefore, offers an additional option to obtain specific ophthalmological information for various purposes. PMID- 9738385 TI - [Intermittent vision disorder caused by mobile vitreous body structure]. PMID- 9738386 TI - [Intraocular interventions in eyes with rubeosis iridis and neovascular glaucoma]. PMID- 9738387 TI - [Theophylline and inhaled n corticosteroids in asthma: a reliable combination]. PMID- 9738388 TI - [Comparison of addition of theophylline to inhaled steroid with doubling of the dose of inhaled steroid in asthma]. AB - The anti-asthmatic effect of theophylline may supplement those of inhaled steroids in asthma. The aim of the present trial was to study how the addition of theophylline compares to doubling the dose of inhaled steroid in asthmatics who remain symptomatic on beclomethasone dipropionate (BDP) 400 micrograms/day. The trial was designed as a randomized, double-blind, parallel-group study in several European countries. 69 patients were treated for 6 weeks with theophylline plus BDP 400 micrograms/day, compared to 64 patients treated with BDP 800 micrograms/day. The mean +/- SD serum theophylline concentration was 10.1 +/- 4.2 mg/l. Lung function measurements were made throughout the study and patients kept daily records of peak expiratory flow rate (PEF), symptoms and salbutamol usage. Forced expiratory volume in one second and PEF at week 6 were significantly increased by both treatments (p < 0.01). PEF variability was reduced by about 30% in both groups. There were significant improvements in asthma symptoms and rescue medication use (p < 0.001). There were no significant differences between the treatment groups. The study demonstrated clinical equivalence of theophylline plus beclomethasone dipropionate 400 micrograms/day and beclomethasone dipropionate 800 micrograms/day in patients whose asthma is not controlled on beclomethasone dipropionate 400 micrograms/d. The results support the use of theophylline as steroid-sparing agent. The combination of low-dose inhaled steroid plus theophylline is a suitable treatment for moderate asthma. PMID- 9738389 TI - [Aneurysm of the aortic arch: presenting as mediastinal tumor]. AB - We report the case of a 62-year-old woman who presented with severe abdominal pain. The routine chest x-ray showed an anterior mediastinal mass measuring approximately 10 cm in diameter. CT-Scan and angiography demonstrated an aneurysm of the aortic arch compromising the left pulmonary artery and main bronchus. An additional aneurysm of the abdominal aorta and right iliac artery was seen, which apparently led to the abdominal symptoms. Coronary angiography revealed coronary artery disease. The aortic arch aneurysm was treated by interposition of a vascular graft. Aorto-coronary bypass grafts were implanted. The abdominal aneurysm was resected in a staged approach after recovery from the first operation. PMID- 9738390 TI - [Dyspnea and lung function. Results of a multicenter study]. AB - In seven pneumological centres 266 patients with different pneumological diseases were investigated. After having clarified several questions regarding the severity of the dyspnoea, cough intensity and the volume of sputum, as well as basic clinical investigation and after an x-ray of the thorax, the diagnosis was arrived at. Subsequently the lung function investigation with the flow-volume curve (including IVC, FVC, PEF, FEV1, MEF50%) and the body plethysmographic Rt and IGV were carried out. Different quality control procedures at and between the different centres ensured comparable results. All centres agreed to using methods well compatible with each other. The question as to which kind of parameters of lung function would agree best with the amount of the dyspnoea, was resolved. The causes for the large scatter of the results are described. Cough and sputum exercise an influence even on the degree of dyspnoea, but not by deteriorating the lung function. The results are shown for the entire collective (Part I) in respect of the different diagnoses (Part II). With different diagnosis the same significant correlations exist but the curves are positioned at different levels of the coordinate system. CONCLUSION: Significant correlations exist between the dyspnoea scale and function parameters. There are individual differences between the dyspnoea scale and disturbances of the function parameters. Carefully performed lung function analyse definitely important in any case. PMID- 9738391 TI - [Antioxidative and clinical effects of high-dose N-acetylcysteine in fibrosing alveolitis. Adjunctive therapy to maintenance immunosuppression]. PMID- 9738392 TI - [German Society of Pneumology: recommendations for diagnosis and therapy of sarcoidosis]. PMID- 9738393 TI - [Report on the current knowledge of Vienna primary school teachers about bronchial asthma in children]. AB - PURPOSE: This study was performed to examine the causes, triggers and therapy of bronchial asthma in a statistically relevant group of teachers at primary schools in Vienna. Furthermore, it was intended to investigate the correlation between their knowledge and their approach in respect of the management of asthmatic pupils. METHODS: 1054 (80.4%) of 1311 questionnaires were returned and evaluated. Five items were investigated: "general knowledge", "symptoms and triggers"; "exercise", "treatment" and "individual experience". Statistical analysis was performed by using counting statistics. For the correlation of items, Spearman correlation coefficients and Wilcoxon's test were used. RESULTS: The teachers in primary schools showed a good basic knowledge of asthma and its symptoms. Poor understanding was found with regard to the medical treatment and trigger factors of asthma; only 34% of the teachers knew that playing games in cold wind may provoke an exacerbation of asthma; less than half of the teachers (45%) were aware of the fact that an asthma attack can be prevented by prophylactic treatment. A significantly positive correlation was found between previous instruction on asthma and its management, the degree of individual experience, and the correct belief that asthmatic children should be encouraged to fully participate in school sports and activities (p = 0.001). Most of the teachers (94%) felt the lack of sufficient information. Only 2% had received proper instruction on asthma and revealed a significantly better knowledge of all items (p = 0.0001). CONCLUSION: We suggest that teachers at primary school receive further instruction on asthma, especially regarding its practical aspects. PMID- 9738394 TI - [Pulmonary hemosiderosis and gastroesophageal reflux in an infant]. AB - Pulmonary hemosiderosis (PH) has been described in association with a variety of immunological and non-immunological diseases. It is characterised by iron deficiency anaemia, hemoptysis and diffuse pulmonary infiltrates based on recurrent intraalveolar hemorrhages. We present the case of a child with pulmonary hemosiderosis and a pathological gastroesophageal reflux activity. The child suffered from recurrent anaemic episodes the age of three months (hemoglobin level up to 5.4 g/dl). The symptoms decreased after removal of the gastroesophageal reflux and accompanying steroid therapy. There has been one relapse of pulmonary hemorrhage seven months later (hemoglobin level 6.1 g/dl). Since then the patient has been in good general condition and the steroid was slowly reduced. No more anaemic episodes occurred. We discuss a possible association of pathological gastroesophageal reflux activity in pulmonary hemosiderosis. PMID- 9738395 TI - [Large, multilocular thymus cyst with compression of the left lung]. AB - Thymic cysts are rare, generally asymptomatic lesions, which in most cases are discovered incidentally on the chest x-ray and are localized in the anterior compartment of the mediastinum or in the neck. We report the case of a 41 year old man, who developed a compression of the left lung and a protuberance of the left chest wall as a result of multilocular thymic cyst with an exceptional volume of 2 litres. PMID- 9738396 TI - [Detection of mutations of the K-ras gene in condensed breath of patients with non-small-cell lung carcinoma (NSCLC) as a possible noninvasive screening method]. PMID- 9738398 TI - [Operationalized psychodynamic diagnosis in childhood and adolescence--a contribution to quality assurance]. AB - Adopting a therapeutic view on diagnosis in child and adolescent psychiatry a psychodynamic diagnostic manual seems to be necessary, that may provide reliability in the assessment of intrapsychic processes. Multiaxial diagnosis of children's and adolescent's psychiatric disturbances may thus be completed with additional psychodynamic assessment procedures. In adults an operationalized procedure of psychodynamic assessment (OPD) already exists. In child and adolescent psychiatry a working group for psychodynamic diagnostic (OPD-KJ) has been founded. In 4 areas (subjective experiences of disturbances, relationships, intrapsychic conflicts and psychic structure) diagnostic manuals are being worked out. Fundamental questions of developmental psychopathology and diagnostic procedures have to be addressed. The basic proposition takes into account that children should not be psychodynamically compared with adults, but normally have a basic capacity to adapt to environment on a high level of psychic organisation in all developmental stages. Reports of the ongoing work are being given. PMID- 9738397 TI - [Unusual metastasis of a malignant pleural mesothelioma]. AB - Usually malignant pleural mesothelioma causes pain and dyspnoea due to local invasion of the chest wall and compression of the lung. Distant metastases rarely cause symptoms. We report on a patient with an epithelial subtype of malignant pleural mesothelioma who presented himself after chemo- and immunotherapy with shortness of breath and loss of weight due to a temporo-mandibular joint and a new nodular shadow in the contralateral lung. The prior diagnosis of an epithelial subtype of pleural mesothelioma was confirmed histologically in a pleural biopsy as well as in the resected orofacial metastases. PMID- 9738399 TI - [Correlation of scope and content in treatment of dyssocial children and adolescents. A psychoanalytic study of disordered symbolization processes]. AB - The article discusses the significance of a holding therapeutic environment for the processes of symbol development. Using epistomological psychoanalytic concepts it demonstrates, through a series of examples, how semiotic progression can be facilitated or how, through insufficient handling of frame guiding therapeutic semiotic regression--in the sense of a "reversal of alpha function"- can be triggered, with a resultant increase in symptomatic behavior. The distinction is drawn between three different levels of mental functioning: At the deepest level, behavior is controlled according to somatic, autoregulative and homoeostatic principles. The second level is concerned with mental functioning, such as determines behavior of individuals within a group and which is influenced by symbol systems and affective automatisms, while obeying a type of "symmetrisation-mechanism." Only at the third level, we are concerned with psychic spaces and the thinking subject, who is capable of symbolic interaction with both the internal and external world. PMID- 9738400 TI - [Cyclical vomiting as the leading symptom of regression]. AB - A 17-year old boy with primary mental retardation of unknown aetiology and cyclic vomiting syndrome (CVS) is reported. Missing an organic cause of the episodes therapeutic procedures remained symptomatic for a 15 years time period. The report focusses the CVS as a regressive mechanism triggered by a mental retardation of the child which permanently disturbed the interaction between parents and child. Psychotherapeutic approach was successful in the following respects: The advantage by illness to vomit at home was continuously reduced by admitting the boy to a hospital at every ongoing attack. Hypnotherapeutic techniques were able to substitute the "malign" regression (CVS) successively by inducing a "benign" kind of regression. Family therapy could induce new and more intense non-verbal patterns of communication which where hitherto unknown in this family. PMID- 9738402 TI - [Aspects of group therapy with sexually abused children--symbolic processing and construction of the initial phase]. AB - This study deals with aspects of group therapy with traumatized children. Various examples from a children's therapy group with sexually abused girls at the age of nursery and primary school enlighten the theoretical explanations. With a reflexion about traumatization by sexual abuse as a starting point a catalogue of therapeutical ends is set up including a methodic realization in exemplary form. A central point is marked by reflexions on the featuring of the initial phase: according to the high level of fear in the children and their minimal abilities to set up limits, there has to be a procedure of absolutely clear structuring; considerations on means establishing security are being focussed. Moreover, this study describes the use of forms of coping by symbols. With the help of theoretical considerations on the role of symbolic function and the games children play the use of the former is explained. Concrete proceeding and the efficacy of coping by symbols are made obvious throughout a treasure of examples. The central motive of this study is to give concrete impulses for practical work with traumatized children and not to discover new continents through scientific research. PMID- 9738401 TI - [Computer in child psychotherapy--on the use of computer games in child guidance counseling]. AB - The article informs about the implementation and the handling of computer games for children and juveniles in psychotherapy. The games in use and the particular therapeutical techniques are described, combined with a report of special ego functions, which can be trained by the use of computer games. Some short examples show the computer-aided treatment in context with the developmental and personality problems of juvenile clients. PMID- 9738403 TI - [Epidemiological chronicle: evidence of change in patterns of infectious diseases]. PMID- 9738404 TI - [Infectious diseases in Poland in 1996]. PMID- 9738405 TI - [Measles in 1996]. PMID- 9738406 TI - [Whooping cough in 1996]. PMID- 9738407 TI - [Scarlet fever in 1996]. PMID- 9738408 TI - [Epidemic parotitis: mumps in 1996]. PMID- 9738409 TI - [Influenza in 1996]. PMID- 9738410 TI - [Rubella in 1996]. PMID- 9738411 TI - [Cerebrospinal meningitis and encephalitis in 1996]. PMID- 9738412 TI - [Salmonellosis in 1996]. PMID- 9738413 TI - [Bacterial dysentery in 1996]. PMID- 9738414 TI - [Poisoning and food poisoning in 1996]. PMID- 9738415 TI - [Botulism in Poland in 1996]. PMID- 9738416 TI - [Poisonings caused by plant protection agents in 1996]. PMID- 9738417 TI - [Hepatitis B virus in 1996]. PMID- 9738418 TI - [Hepatitis B vs. Hepatitis non-B in 1996]. PMID- 9738419 TI - [Tetanus in 1996]. PMID- 9738421 TI - [Brucellosis in 1996]. PMID- 9738422 TI - [Trichinosis in 1996]. PMID- 9738420 TI - [Rabies in Poland in 1996]. PMID- 9738423 TI - [Taenia infections in 1996]. PMID- 9738424 TI - [Malaria in Poland in 1996]. PMID- 9738425 TI - [AIDS and HIV infections in 1996]. PMID- 9738426 TI - [Prevalence of HBV and HCV infection markers in patients with sexually transmitted diseases]. AB - Sera of 125 patients with sexually transmitted diseases (syphilis, gonorrhoea, chlamydiosis, HPV and HIV infections) were investigated for presence of 3 markers of HBV infection; they were found in 41 (33%) patients. Anti-HBc was present in sera of 35 (28%) patients, in 3 of them antigen HBs was found and in 28 anti-HBs was found as well. Antigen HBs alone was present in sera of 6 other patients but they were not reactive in test for anti-HBc. Moreover in this group of 125 patients anti-HCV were discovered in 4 (3%); in 3 of them occurrence of markers of HBV infection was found. PMID- 9738427 TI - [Screening lab tests of Chlamydia trachomatis. Do they show declining trend of the infections?]. AB - A study on the frequency of Chlamydia trachomatis (Ct) urogenital infection comprised 2393 patients (1,197 women and 1,196 men), aged 18-69 years. Urethral and cervical specimens were tested using immunofluorescent (DFA) for symptomatic and immunoenzymatic techniques (EIA, IMx, Vidas-ELFA) for asymptomatic patients. Basing on the tests results (1993-1996) chlamydiosis was diagnosed in 323 (13.5%) cases, 169 (14.1%) women and 154 (12.8%) men, aged 20-40 yrs. The percentage of positive results determined by EIA, IMx or Vidas were similar (8.2-4.2%, 3.0 2.0%, 5.4-3.4%, respectively) but lower than that by DFA test (38.9-13.2%). Over the last few years the number of patients screened for Ct, as well as the incidence of Ct infection, has decreased (5.6% in 1996). The data were compared to those from 1986-1992. PMID- 9738428 TI - [Sterile packaging]. AB - After decontamination, cleaning, maintenance and functional testing, sterilized items must be packed suitably. The package must protect sterilized items against microbial contamination during removal from the sterilising chamber, and during storage or transport until use. The sterilized material must be packaged in suitable packaging material in accordance with the sterilising method. Sterilization packaging must conform to standards. There are not Polish standards for sterile packaging of medical devices. The article give general guidelines about: properties of sterile packaging, various national standards and European Standard, the packaging materials, general compatibility with the sterilization process which is intended to be used, general compatibility with the package forming process, and shelf life considerations. PMID- 9738429 TI - [The possibility of transmission of Helicobacter pylori by gastroscopes]. PMID- 9738430 TI - [Superantigens: new data]. AB - The paper pertains novel data on action of bacterial and viral superantigens on the immune system. Against the background, new morbid conditions have been described, which may result from the action of superantigens. In the context of superantigen action on the immune system, antigen pathways in the system have been described. The review has been based on laboratory results obtained during recent 5 years in various scientific centers in the world. PMID- 9738431 TI - [The penicillin susceptibility of Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis and Staphylococcus aureus in persons suffering from upper respiratory tract infections]. AB - The susceptibility to penicillin was tested in 293 strains of S. aureus, 162 strains of H. influenzae, 60 strains of M. catarrhalis and 77 strains of S. pneumoniae, isolated from persons suffering from upper respiratory tract infections. Penicillin susceptibility testing has been performed by the disc diffusion technique. Beta-lactamase production in the penicillin-resistant strains of H. influenzae, M. catarrhalis and S. aureus was detected with nitrocefin impregnated disc. In beta-lactamase positive strains, the susceptibility to amoxacillin/clavulanic acid (AMC) was determined. Nearly half (46.3%) of H. influenzae strains was resistan to ampicillin. In the remaining bacteria, the percentage of resistance was 2.6% for S. pneumoniae, 82.4% for M. catarrhalis and 88.1% for S. aureus strains. All strains resistant to penicillin (ampicillin) produced beta-lactamase and were AMC sensitive. PMID- 9738432 TI - [Central anti-typhus committee (August 1, 1919-March 5, 1920)]. AB - During the first period of the independence of Poland, just after the first World War (1919-1921), the fight against ramped epidemics of acute infectious diseases was a main priority. The most important task of the Ministry of Public Health was the organization of the control of infectious diseases, and especially typhus. In 1919, the government named the Central Anti-Typhus Committee. However, the Committee was a collective institution without necessary competences and resources and its efforts to control typhus epidemics have been unsuccessful. At the end of 1919, there was a dramatic spread of typhus especially in the eastern parts of the country, still suffering from the war. In March 1920, the Central Anti-Typhus Committee has been replaced by the Extraordinary Epidemic Commissariat which has been endowed with special, emergency staff and resources. The Commissariat was headed by Prof. Emil Godlewski, who was soon nicknamed "tyhus tsar"; for the most affected regions competent regional commissars were appointed. The legal ground of the Extra ordinary Epidemic Commissariat was the parliament bill on 14th July 1920. This body took over from the Public Health Minister responsibilities for the fight against epidemics. The work of Commissariat had stopped the epidemics and improved epidemical status of Poland which could be seen from 1923 onwards. PMID- 9738433 TI - Industry's perception and use of occupational exposure limits. AB - Market research was carried out to determine industry's perception of occupational exposure limits (OELs) and the extent to which they influence the selection of measures to control exposure. Telephone interviews were carried out with 1000 randomly selected users of chemicals, 400 from establishments with some use of chemicals and 600 from establishments with chemicals in daily use. 150 interviews were also carried out with Trade Union Health and Safety Representatives. The interviews covered basic information on chemicals used, sources of information, risk reduction measures used and understanding of COSHH and OELs. Most respondents came from firms with 10 employees or less (75%, all user group; 57%, heavy user group), closely reflecting the profile of British industry. In contrast, most (77%) Trade Union Health and Safety Representatives came from firms with at least 100 employees. Respondents in the all user group were drawn from across the whole range of industrial activity, whereas the heavy users were concentrated in manufacturing. The results showed that in making decisions on what control measures to use most users rely heavily on information from suppliers and personal experience and rather less on information from sources such as Trade Associations and HSE. Most respondents reported that steps were taken to protect employees. The use of personal protective equipment featured highly, followed by process controls. Little consideration was given to the possibility of substitution. Awareness of COSHH was limited with 65% of the all user group and 53% of the heavy user group being unaware of any legal requirements for establishments which manufacture or work with chemicals. Awareness of OELs was similarly limited with 19% of the all user group and 32% of the heavy user group having any real understanding. The results from Trade Union Representatives showed that overall they are somewhat better informed than chemical users in the small firms surveyed. PMID- 9738434 TI - An introduction to a UK scheme to help small firms control health risks from chemicals. AB - The Control of Substances Hazardous to Health Regulations 1994 (COSHH), provide the main British legislation to protect against health risks arising from hazardous substances used at work. Under the regulations, employers have a duty to carry out a suitable and sufficient risk assessment and take steps to ensure exposure is adequately controlled. The paper by Topping et al. (1998) concluded that small firms need more basic, readily available advice on how to effectively control hazardous substances. To meet this need the Health and Safety Executive (HSE) and the Advisory Committee on Toxic Substances (ACTS) have developed a new scheme for the UK. It involves a simple system of generic risk assessments to identify appropriate control strategies and a series of control guidance sheets providing good-practice examples of those strategies for common operations. The approach builds on earlier industry risk banding schemes and HSE's general approach to risk assessment and risk management. To help ensure the advice reaches small firms, HSE is seeking to involve key intermediaries in its dissemination. This paper describes the rationale for the new UK scheme, how it sits in the legal framework, and proposals for its dissemination. The papers by Brooke (1998) and Maidment (1998) set out in detail the technical basis for the scheme. PMID- 9738435 TI - A UK scheme to help small firms control health risks from chemicals: toxicological considerations. AB - The UK has developed a simple scheme to provide practical control advice to small and medium-sized enterprises (SMEs), to assist them in their risk assessments and risk management decisions. This scheme makes use of toxicological hazard information indicated by R-phrases assigned under the European Union (EU) classification system, to assign substances to hazard bands. In the UK scheme, the allocation of substances to hazard bands according to R-phrases has taken into account three key factors: whether or not the toxicological endpoint has an identifiable dose threshold; the seriousness of the resultant health effect; and the relative exposure levels at which toxic effects occur. Based on all these considerations, R-phrases have been allocated to hazard bands within the scheme. An evaluation exercise has been undertaken, to compare the output of the scheme with established health-based occupational exposure limits, for more than 100 substances. The results of this exercise demonstrate that as far as possible, the scheme recommends control strategies which should provide adequate control. This scheme is potentially a very powerful means of helping SMEs adequately control chemical health risks in the workplace. Since it utilises the EU-agreed classification system, the scheme can be applied to any substance supplied and used in the workplace and it may also be used internationally. PMID- 9738437 TI - Information needs of workers. AB - In Britain, the law places duties on employers and suppliers to provide information to ensure health and safety of employees, so far as is reasonably practicable, and there are regulations for the appointment of employees' safety representatives which employers are required to consult. A recent survey by HSE has shown that suppliers are the most important source of information on substances. However, the flow of information is often obstructed by barriers between the suppliers and the companies, and within organisations. Safety representatives, where they exist, are often better informed than employers, and in companies with safety representatives accident rates are lower. Information from suppliers can be inappropriate for the end use, and the goal-setting approach which has dominated in recent years may not help the non-expert employer. We welcome HSE's move to more specific control guidance for chemicals. PMID- 9738436 TI - Occupational hygiene considerations in the development of a structured approach to select chemical control strategies. AB - This paper explains the occupational hygiene basis of a new scheme to help small firms control the health risks from supplied chemicals. The scheme groups hazard information and the potential for a material to become airborne into bands and, from this information, predicts the control strategy necessary to ensure that the hazardous substance is used safely. To do this a simple model based upon an empirical approach to risk assessment and risk management has been developed. This work was undertaken in a working group established by the Health and Safety Commission's Advisory Committee on Toxic Substances. PMID- 9738438 TI - Target speed alone influences the latency and temporal accuracy of interceptive action. AB - When intercepting a mobile object or an apparent movement, participants show a temporal bias. They are in advance when dealing with a slow-moving stimulus and late with a fast-moving one. We studied participants intercepting an apparent movement by sliding a disk on a table. Using a fast and a slow stimulus speed, we varied three factors: duration of presentation of the stimulus, distance covered by the stimulus, and speed context (constant or varied) of stimulus presentation. In addition to the temporal bias, spatial accuracy and kinematic measures were collected. The temporal bias created by speed was evident across all three factors. Speed, in addition to strongly determining the temporal bias, significantly affected the throwing strategy adopted by the participants, as revealed by latency, movement time, and disk trajectory duration. PMID- 9738439 TI - Response preparation and control of movement sequences. AB - Two experiments investigated the response complexity effect using elbow extension/flexion movements. In the first experiment, RT for an extension movement was significantly less than RT for an extension/flexion movement. However, this difference in RT was not evident when participants were asked to pause at the reversal of the extension/flexion for approximately 260 ms. The second experiment manipulated the duration of the pause between these movements and also measured the electromyographical (EMG) activity of the triceps and biceps muscles. When the pause was reduced to 75 ms participants were not able to program the flexion portion of the movement at the reversal, forcing them to preprogram this movement; hence, increasing their premotor reaction time. PMID- 9738440 TI - Fe65 and the protein network centered around the cytosolic domain of the Alzheimer's beta-amyloid precursor protein. AB - A distinctive tract of all the forms of Alzheimer's disease is the extracellular deposition of a 40-42/43 amino acid-long peptide derived from the so-called beta amyloid precursor protein (APP). This is a membrane protein of unknown function, whose short cytosolic domain has been recently demonstrated to interact with several proteins. One of these proteins, named Fe65, has the characteristics of an adaptor protein; in fact, it possesses three protein-protein interaction domains: a WW domain and two PID/PTB domains. The interaction with APP requires the most C-terminal PID/PTB domain, whereas the WW domain is responsible for the interaction with various proteins, one of which was demonstrated to be the mammalian homolog of the Drosophila enabled protein (Mena), which in turn interacts with the cytoskeleton. The second PID/PTB domain of Fe65 binds to the CP2/LSF/LBP1 protein, which is an already known transcription factor. The other proteins interacting with the cytosolic domain of APP are the G(o) heterotrimeric protein, APP-BP1 and X11. The latter interacts with APP through a PID/PTB domain and possesses two other protein-protein interaction domains. The small size of the APP cytodomain and the overlapping of its regions involved in the binding of Fe65 and X11 suggest the existence of competitive mechanisms regulating the binding of the various ligands to this cytosolic domain. In this short review the possible functional roles of this complex protein network and its involvement in the generation of Alzheimer's phenotype are discussed. PMID- 9738441 TI - The distance between the 3'-pyrimidine-rich tract and the AUG codon modulates internal initiation of translation of hepatitis A virus RNA. AB - Protein synthesis directed by hepatitis A virus (HAV) RNA is mediated by a mechanism involving the recognition of internal sequences. Two in-frame AUG codons initiate the long open reading frame (positions 734-736 and 740-742). The extra-cistronic region extending between the uncapped 5'-end and the ORF contains two pyrimidine-rich tracts (PRTs): one 12 nucleotides in length in the close vicinity of the initiator AUG, and a longer one between bases 94 and 140. In order to study the relative contribution of these elements to the process of internal initiation of translation, cDNA representations of the 5'-terminal extra cistronic region of HAV RNA were inserted in the intergenic region of the bi cistronic plasmid pSV-GH/CAT, between the genes encoding the human growth hormone (GH) and the bacterial enzyme chloramphenicol acetyltransferase (CAT), and following transfection of COS-1 cells, the transient expression of both genes was quantified. The importance of the 3'-PRT appeared to be strongly influenced by the length of the 'spacer' sequence extending between this structure and the translation initiation site: placed 45 nucleotides upstream from the initiator codon of a reporter gene, its integrity was stringently required for initiation to occur. Bringing the length of the 'spacer' back to its actual size in HAV RNA (i.e. 11 or 17 nt) reduced considerably the overall rate of internal initiation of translation, and the relative contribution to this process of the 3'-PRT became marginal. Concomitantly, the importance of the functional domains previously identified in the 5'-PRT fluctuated: while integrity of domain 100-106 was always stringently required for initiation to occur, the activity of domain 113-118 paralleled that of the 3'-PRT, and the opposite applied to domain 121 126, whose contribution became relevant only after switching off the 3'-PRT. Systematic mutations introduced in the 'spacer' sequences suggest that the length of this region may be responsible for the down regulation of translation of HAV RNA and, possibly, for its lengthy replication cycle. PMID- 9738442 TI - Operation of glyoxylate cycle in halophilic archaea: presence of malate synthase and isocitrate lyase in Haloferax volcanii. AB - The occurrence of the glyoxylate cycle has not previously been demonstrated in any of the Archaea. In halophilic archaea, only isocitrate lyase activity has been detected. The halophilic archaeon Haloferax volcanii was tested for the presence of the other key enzyme of this pathway, malate synthase. High activities of this enzyme were detected when the carbon source was acetate. Both glyoxylate cycle key enzymes, isocitrate lyase and malate synthase, from Hf. volcanii were purified and characterized. PMID- 9738443 TI - Electrical current generation and proton pumping catalyzed by the ba3-type cytochrome c oxidase from Thermus thermophilus. AB - Several amino acid residues that have been shown to be essential for proton transfer in most cytochrome c oxidases are not conserved in the ba3-type cytochrome c oxidase from the thermophilic eubacterium Thermus thermophilus. So far, it has been unclear whether the Th. thermophilus ba3-type cytochrome c oxidase can nevertheless function as an electrogenic proton pump. In this study, we have combined charge translocation measurements on a lipid bilayer with two independent methods of proton pumping measurements to show that enzymatic turnover of the Th. thermophilus cytochrome c oxidase is indeed coupled to the generation of an electrocurrent and proton pumping across the membrane. In addition to a 'vectorial' consumption of 1.0 H+/e- for water formation, proton pumping with a stoichiometry of 0.4-0.5 H+/e- was observed. The implications of these findings for the mechanism of redox-coupled proton transfer in this unusual cytochrome c oxidase are discussed. PMID- 9738444 TI - Engineering of solvent-exposed loops in Escherichia coli beta-galactosidase. AB - The Escherichia coli beta-galactosidase is a high molecular mass tetrameric enzyme extensively used as a molecular marker. Despite its proven utility as a partner in fusion proteins, previous attempts to generate insertional mutants rendered inactive or poorly active enzymes, hampering its further engineering for the construction of multifunctional enzymes. We have explored several solvent exposed loops on the tetramer, namely those spanning residues 246-254, 271-287, 578-584, 770-773, and 793-806, as acceptor sites to accommodate functional protein segments on the surface of active beta-galactosidase enzymes. An RGD containing antigenic peptide positioned in these sites interacts efficiently with specific monoclonal antibodies as well as target integrins on the surface of mammalian cells. The resulting chimeric enzymes are soluble, stable, produced in high yields and enzymatically active. Moreover, the identified insertion sites could be appropriated for the design of promising beta-galactosidase-based molecular sensors. PMID- 9738445 TI - Expression of pI(Cln) in Escherichia coli gives a strong tolerance to hypotonic stress. AB - We amplified the coding region DNA sequence from a rat renal pI(Cln) cDNA by PCR and expressed the protein in Escherichia coli cells. The cells were exposed to hypotonic conditions followed by spreading them onto LB plates for subsequent colony survival assay. The present study demonstrated that the cells expressing pI(Cln) exhibit a strong resistance to hypotonic stress. Moreover, the resistance was specifically inhibited by extracellular ATP and some anion channel inhibitors. These findings indicate that the expression of pI(Cln) directly confers tolerance to hypotonic stress, and pI(Cln) is concluded to be an important molecule for cell-volume regulation. PMID- 9738446 TI - Stress-induced expression of transcriptional repressor ICER in the adrenal gland. AB - Second messenger cyclic AMP plays a central role in signalling within the hypothalamo-pituitary-adrenal (HPA) axis. Changes in gene expression are central to long-term adaptations made in response to stress in the adrenal gland. Here we demonstrate that expression of the cAMP inducible transcriptional repressor, ICER (Inducible cAMP Early Repressor), is rapidly and powerfully induced in response to surgical stress in the rat adrenal gland. Hypophysectomisation blocks stress induced ICER expression. Finally we demonstrate that injection of the pituitary hormone ACTH (Adrenocorticotropin Hormone) induces robust ICER expression in the adrenal cortex. Thus, induction of the transcriptional repressor ICER is coupled to the HPA axis response to stress. PMID- 9738447 TI - Ligand-induced endocytosis of the asialoglycoprotein receptor: evidence for heterogeneity in subunit oligomerization. AB - The hepatic asialoglycoprotein receptor, a noncovalent hetero-oligomer of two subunits, is a constitutively cycling endocytic receptor. However, the ligand asialoorosomucoid caused downregulation of up to 40% of surface binding sites and a twofold increase in internalization rate. This was not the result of receptor crosslinking, since monovalent ligands had the same effect. Ligand binding thus appears to transmit a signal to the cytosolic portion of the receptor not unlike in signaling receptors. The two subunits were endocytosed at different average rates lower than that of ligand, indicating heterogeneity in oligomer formation and potentially in ligand specificity. PMID- 9738448 TI - Multiple crystal forms of hexokinase I: new insights regarding conformational dynamics, subunit interactions, and membrane association. AB - Hexokinase I is comprised of homologous N- and C-terminal domains, and binds to the outer membrane of mitochondria. Reported here is the structure of a new crystal form of recombinant human hexokinase I, which complements existing crystal structures. Evidently, in some packing environments and even in the presence of glucose and glucose 6-phosphate the N-terminal domain (but not the C terminal domain) can undergo oscillations between closed and partially opened conformations. Subunit interfaces, present in all known crystal forms of hexokinase I, promote the formation of linear chains of hexokinase I dimers. Presented is a model for membrane-associated hexokinase I, in which linear chains of hexokinase I dimers are stabilized by interactions with mitochondrial porin. PMID- 9738449 TI - Intracellular free calcium concentration in human taste bud cells increases in response to taste stimuli. AB - We examined changes of intracellular free calcium concentration [Ca2+]i elicited by taste stimuli of sucrose, denatonium and NaCl in the taste buds of seven human fungiform papillae. In one taste bud we observed an increase in [Ca2+]i induced by only NaCl. In another bud an increase of [Ca2+]i in response to both NaCl and sucrose was found. The Ca2+ responses to NaCl and sucrose occurred in differential areas within the one taste bud. In the other five fungiform papillae [Ca2+]i was not changed by the taste stimuli. These results suggest that an increase of [Ca2+]i participates in taste transduction mechanisms for sucrose and NaCl, and that taste cells in one taste bud may respond to differential stimuli. PMID- 9738450 TI - An intronic promoter controls the expression of truncated human gamma glutamyltransferase mRNAs. AB - We have identified and characterized a genomic DNA fragment containing the coding sequences corresponding to the human gamma-glutamyltransferase type 1 mRNA. The coding part of the gene spans over 16 kb and comprises 12 exons and 11 introns exhibiting a similar organization as for the mouse and rat GGT genes. The exons 1 7 encode the heavy subunit whereas exons 8-12 which encode the carboxy-terminal part of the heavy subunit (exon 8) and the light subunit are clustered in a 1.6 kb BglII fragment. Exons 7 and 8 are separated by a 3.9-kb intron containing in its 3' part the sequences corresponding to the 5'-UTRs of the truncated GGT mRNAs described for human lung. Sequence analysis upstream this transcribed region exhibited putative promoter sequences and after transient transfection significant promoter activities were measured in V79 lung fibroblasts and KYN-2 hepatoma cells but not in A2780 ovarian cells. This specificity disappeared when only 550 bp upstream the transcription start site were used as promoter. These results argue for a promoter of truncated GGT mRNAs in intron 7, specifically regulated in human tissues. PMID- 9738451 TI - ATP synthesis by the F1Fo ATP synthase of Escherichia coli is obligatorily dependent on the electric potential. AB - The H+-translocating F1Fo ATP synthase of Escherichia coli was purified and reconstituted into proteoliposomes. This system catalyzed ATP synthesis when energized by an acid/base transition (pHin = 5.0; pHout = 8.3) with succinate, malonate or maleinate but not with MES as the acidic buffer. Under these experimental conditions an electric potential of 125-130 mV is generated by the diffusion of succinate, probably the monoanionic species, whereas with MES buffer the measured potential was at background level (approximately 5 mV). ATP was also synthesized at pH 7.2 in the absence of a delta pH by applying a K+/valinomycin diffusion potential. The rate of ATP synthesis increased with the potential in an exponential manner with an inflection point at about 70 mV. We conclude from these results that delta pH and delta psi are kinetically unequivalent driving forces for ATP synthesis by the E. coli ATP synthase and that delta psi is a mandatory force for this synthesis. The significance of these findings for the mechanism of ATP synthesis in general is discussed. PMID- 9738452 TI - ATP binding site of P2X channel proteins: structural similarities with class II aminoacyl-tRNA synthetases. AB - The extracellular loop of P2X channel proteins contains a sequence stretch (positions 170-330) that exhibits similarities with the catalytic domains of class II aminoacyl-tRNA synthetases as shown by secondary structure predictions and sequence alignments. The arrangement of several conserved cysteines (positions 110-170) shows similarities with metal binding regions of metallothioneins and zinc finger motifs. Thus, for the extracellular part of P2X channel proteins a metal binding domain and an antiparallel six-stranded beta pleated sheet containing the ATP binding site are very probable. The putative channel forming H5 part (positions 320-340) shows similarities with the enzyme motif 1 responsible for aggregation of subunits to the holoenzyme. PMID- 9738453 TI - Paracrine regulation of talin mRNA expression by androgen in human prostate. AB - Androgens are essential for normal prostate physiology and are intimately associated with the growth and progression of prostate cancer. However, few androgen regulated genes in the prostate have been identified. Using the mRNA differential display technique a 164-bp cDNA fragment was identified as being androgen regulated in the human prostate. Nucleotide sequence analysis of this fragment revealed 84% homology with the gene encoding the cytoskeletal protein talin. Confirmation of the androgen regulation of this gene was carried out using Northern analysis. Primary prostatic stromal cells treated with conditioned medium (CM) from androgen-treated primary prostatic epithelial cells showed an approximate 2-fold reduction in talin mRNA levels compared with stromal cells treated with CM from epithelial cells not exposed to androgens. Expression of talin mRNA in human prostatic tissue was confirmed by in situ hybridisation. The highest levels of expression were present in the epithelial cells, with lower levels of expression in the stroma. Thus, androgen regulation of talin expression may play a role in normal and/or aberrant growth and development of the prostate. PMID- 9738454 TI - The hexokinase 2 protein participates in regulatory DNA-protein complexes necessary for glucose repression of the SUC2 gene in Saccharomyces cerevisiae. AB - The HXK2 gene plays an important role in glucose repression in the yeast Saccharomyces cerevisiae. Recently we have described that the HXK2 gene product, isoenzyme 2 of hexokinase, is located both in the nucleus and in the cytoplasm of S. cerevisiae cells. In this work we used deletion analysis to identify the essential part of the protein-mediating nuclear localisation. Determinations of fructose-kinase activity and immunoblot analysis using anti-Hxk2 antibodies in isolated nuclei, together with observations of the fluorescence distribution of Hxk2-GFP fusion protein in cells transformed with an HXK2::gfp mutant gene, indicated that the decapeptide KKPQARKGSM, located between amino acid residues 7 and 16 of hexokinase 2, is important for nuclear localisation of the protein. Further experimental evidence, measuring invertase activity in wild-type and mutant cells expressing a truncated version of the Hxk2 protein unable to enter the nucleus, shows that a nuclear localisation of Hxk2 is necessary for glucose repression signalling of the SUC2 gene. Furthermore, we demonstrate using gel mobility shift analysis that Hxk2 participates in DNA-protein complexes with cis acting regulatory elements of the SUC2 gene promoter. PMID- 9738455 TI - Effect of P-chirality of oligo(deoxyribonucleoside phosphorothioate)s on the activity of terminal deoxyribonucleotidyl transferase. AB - Phosphorothioate analogues of oligonucleotides (PS-oligos) of predetermined chirality at the phosphorus atom at each internucleotide linkage have been used as primers for terminal deoxyribonucleotidyl transferase (TdT, EC 2.7.7.31). The enzyme catalyzes efficient elongation of PS primers in which all phosphorothioate internucleotide linkages are uniformly of the [R(P)] configuration, while the presence of the linkage(s) of the [S(P)] configuration significantly decreases or completely inhibits the primer extension. Our results indicate that for the elongation of phosphorothioate oligomers the most important is the internucleotide bond located between the second and the third nucleoside from the 3'-end. The presence of [S(P)] linkage at this position strongly reduces the enzyme activity while the [R(P)] bond allows for effective elongation of the primer. The activity of the enzyme is also influenced by base composition and sequence of phosphorothioate primer as well as the dNTP used for elongation process. PMID- 9738456 TI - The L45 loop in type I receptors for TGF-beta family members is a critical determinant in specifying Smad isoform activation. AB - Transforming growth factor-beta (TGF-beta) and bone morphogenetic proteins (BMPs) signal via distinct type I and type II receptors and Smad proteins. A nine amino acid sequence between kinase subdomains IV and V in type I receptors, termed the L45 loop, has been shown to be important in conferring signalling specificity. We examined the responses of a mutant TGF-beta type I receptor (TbetaR-I) and a mutant BMPR-IB, in which the L45 regions of these two receptors were exchanged. Swapping the four amino acid residues that are different in BMPR-IB for those in TbetaR-I, and vice versa, switched their type I receptor-restricted Smad activation and specificity in transcriptional responses. These studies identify the L45 loop regions in type I receptors as critical determinants in specifying Smad isoform activation. PMID- 9738457 TI - Modification of pigment composition in the isolated reaction center of photosystem II. AB - The pigment content of isolated reaction centers of photosystem II was modified using an exchange protocol similar to that used for purple bacterial reaction centers. With this method, which is based on incubation of reaction centers at elevated temperature with an excess of chemically modified pigments, it was possible to incorporate [3-acetyl]-chlorophyll a and [Zn]-chlorophyll a into photosystem II reaction centers. Pigment exchange has been verified by absorption, circular dichroism and fluorescence spectroscopy, and quantitated by HPLC analysis of pigment extracts. PMID- 9738458 TI - The relationship between synonymous codon usage and protein structure. AB - The hypothesis that synonymous codon usage is related to protein three dimensional structure is examined by investigating the correlation between synonymous codon usage and protein secondary structure. All except two codons in E. coli show the same secondary structural preference for alpha-helix, beta strand or coil as that of amino acids to be encoded by the respective codons, while 17 codons show secondary structural bias in mammalian proteins. The results indicate that there is no significant correlation between synonymous codon usage and protein secondary structure in E. coli, but there is a correlation in mammals. It could be deduced that synonymous codons carry much less structural information in prokaryotes than in eukaryotes due to their divergent evolutionary mechanism. PMID- 9738459 TI - Inhibition of NADPH supply by 6-aminonicotinamide: effect on glutathione, nitric oxide and superoxide in J774 cells. AB - We have examined the integrity of J774 cell nitric oxide (NO) production and glutathione maintenance, whilst NADPH supply was compromised by inhibition of the pentose pathway with 6-aminonicotinamide. In resting cells 6-phosphogluconate accumulation began after 4 h and glutathione depletion after 24 h of 6 aminonicotinamide treatment. Cellular activation by lipopolysaccharide/interferon lambda decreased glutathione by approximately 50% and synchronous 6 aminonicotinamide treatment exacerbated this to 31.2% of control (P < 0.05). In activated cells NO2- production was inhibited by 60% with 6-aminonicotinamide (P < 0.01), and superoxide production by 50% (P < 0.01) in zymosan-activated cells. NADPH production via the pentose pathway is therefore important to sustain macrophage NO production whilst maintaining protective levels of glutathione. PMID- 9738460 TI - Positive feedback between ethanolamine-specific phospholipid base exchange and cytochrome P450 activities in rat liver microsomes. The effect of clofibric acid. AB - The results of the present investigation relate the effects of the nutritional state and administration of clofibric acid (CLA), a hypolipidaemic drug and peroxisomal proliferator, on phosphatidylethanolamine (PE) synthesis in rat liver and fatty acid metabolism. Fasting and CLA treatment of animals causes an increase in the amount of PE in endoplasmic reticulum (ER) membranes and mitochondria, as well as in the PE/phosphatidylcholine (PC) ratio. Moreover, the activity of the ethanolamine-specific phospholipid base exchange (PLBE) enzyme in liver ER membranes of fasted animals was enhanced by 75% in comparison to that of animals fed ad libitum. The effect of CLA treatment was additive to that of starvation; PE synthesis tested in vitro via the Ca2+-sensitive PLBE reaction increased 3-fold in comparison to rats fed ad libitum. This is confirmed by an increased Vmax for the reaction, but the affinity of the enzyme for ethanolamine was not significantly changed. These effects were accompanied by an enhanced expression of cytochrome P450 CYP4A1 isoform and elevated activity of the enzyme upon CLA administration. The stimulatory effect of CLA administration on the efficiency of the ethanolamine-specific PLBE reaction can be explained by elimination of lauric acid, a known inhibitor of de novo PE synthesis, during the course of omega-hydroxylation catalysed by CYP4A1, and by increased expression of the PLBE enzyme. The products of omega-hydroxylation of lauric acid, which are then converted by dehydrogenase to 1,12-dodecanedioic acid, did not significantly affect the in vitro synthesis of PE. PMID- 9738461 TI - Promoter-independent regulation of cell-specific dopamine receptor expression. AB - Here we describe the construction of recombinant adenoviruses expressing dopamine D2 and D4 receptors, and their ability to mediate high levels of heterologous expression in a variety of cell types in vitro and in vivo for at least 7 days post infection. These experiments demonstrated that maximum receptor expression is achieved generally within 24 h and remains constant thereafter. Maximum expression levels were highly variable between cell lines and dependent on infection efficiency and promoter strength. Correction for these two variables revealed differences in relative expression levels between cell lines varying by two orders of magnitude. Our results indicate that in addition to gene transcription, post-transcriptional mechanisms play a dominant role in determining dopamine receptor levels in this system. PMID- 9738462 TI - p53 and the ribosomal protein L5 participate in high molecular mass complex formation with protein kinase CK2 in murine teratocarcinoma cell line F9 after serum stimulation and cisplatin treatment. AB - Using the murine teratocarcinoma cell line F9 we investigated the influence of serum stimulation and cisplatin treatment on the p53, CK2, MDM2 levels. Both treatments led to an increase of p53, though with different kinetics; the other proteins investigated were not affected. We present direct evidence by immunoprecipitation for an association of protein kinase CK2 holoenzyme (alpha2beta2), p53, and the ribosomal protein L5. The results suggest complexes between the CK2 holoenzyme and p53 but also p53/CKbeta complexes. Furthermore we provide evidence for the existence of high molecular mass complexes of CK2 in vivo. This is the first evidence that, under physiological conditions, protein kinase CK2 does not exist solely as a heterotetramer, but predominantly in association with other proteins. PMID- 9738463 TI - The double-stranded RNA-binding domains of Xenopus laevis ADAR1 exhibit different RNA-binding behaviors. AB - We have cloned cDNAs encoding two versions of Xenopus double-stranded RNA adenosine deaminase (ADAR1). Like ADAR1 proteins from other species Xenopus ADAR1 contains three double-stranded RNA-binding domains (dsRBDs) which are most likely required for substrate binding and recognition of this RNA-editing enzyme. Analysis of mammalian ADAR1 identified the third dsRBD in this enzyme as most important for RNA binding. Here we analyzed the three dsRBDs of Xenopus ADAR1 for their in vitro RNA-binding behavior using two different assays. Northwestern assays identified the second dsRBD in the Xenopus protein as most important for RNA binding while in-solution assays demonstrated the importance of the third dsRBD for RNA binding. The differences between these two assays are discussed and we suggest that both the second and third dsRBD of Xenopus ADAR1 are important for RNA binding in vivo. We show further that all three dsRBDs can contribute to a cooperative binding effect. PMID- 9738464 TI - Mutational analysis of caveolin-induced vesicle formation. Expression of caveolin 1 recruits caveolin-2 to caveolae membranes. AB - Caveolae are vesicular organelles with a characteristic uniform diameter in the range of 50-100 nm. Although recombinant expression of caveolin-1 is sufficient to drive caveolae formation, it remains unknown what controls the uniform diameter of these organelles. One hypothesis is that specific caveolin-caveolin interactions regulate the size of caveolae, as caveolin-1 undergoes two stages of self-oligomerization. To test this hypothesis directly, we have created two caveolin-1 deletion mutants that lack regions of caveolin-1 that are involved in directing the self-assembly of caveolin-1 oligomers. More specifically, Cav-1 delta61-100 lacks a region of the N-terminal domain that directs the formation of high molecular mass caveolin-1 homo-oligomers, while Cav-1 deltaC lacks a complete C-terminal domain that is required to allow caveolin homo-oligomers to interact with each other, forming a caveolin network. It is important to note that these two mutants retain an intact transmembrane domain. Our current results show that although Cav-1 delta61-100 and Cav-1 deltaC are competent to drive vesicle formation, these vesicles vary widely in their size and shape with diameters up to 500-1000 nm. In addition, caveolin-induced vesicle formation appears to be isoform-specific. Recombinant expression of caveolin-2 under the same conditions failed to drive the formation of vesicles, while caveolin-3 expression yielded caveolae-sized vesicles. These results are consistent with the previous observation that in transformed NIH 3T3 cells that lack caveolin-1 expression, but continue to express caveolin-2, no morphologically distinguishable caveolae are observed. In addition, as caveolin-2 alone exists mainly as a monomer or homo-dimer, while caveolins 1 and 3 exist as high molecular mass homo-oligomers, our results are consistent with the idea that the formation of high molecular mass oligomers of caveolin are required to regulate the formation of uniform caveolae-sized vesicles. In direct support of this notion, regulated induction of caveolin-1 expression in transformed NIH 3T3 cells was sufficient to recruit caveolin-2 to caveolae membranes. The ability of caveolin-1 to recruit caveolin-2 most likely occurs through a direct interaction between caveolins 1 and 2, as caveolins 1 and 2 are normally co-expressed and interact with each other to form high molecular mass hetero-oligomers containing both caveolins 1 and 2. PMID- 9738465 TI - Human cathepsin X: a novel cysteine protease of the papain family with a very short proregion and unique insertions. AB - A novel cDNA encoding a cysteine protease of the papain family named cathepsin X was obtained by PCR amplification from a human ovary cDNA library. The cathepsin X cDNA is ubiquitously expressed in human tissues and contains an open reading frame of 912 nucleotides encoding a predicted protein of 303 amino acids. All highly conserved regions in papain-like cysteine proteases including the catalytic residues are present in cathepsin X. The mature part of cathepsin X is 26-32% identical to human cathepsins B, C, H, K, L, O, S and W. The cathepsin X sequence contains several unique features: (i) a very short proregion; (ii) a three amino acid residue insertion in a highly conserved region between the glutamine of the putative oxyanion hole and the active site cysteine; and (iii) a second insertion of 15 amino acid residues that can be aligned with the occluding loop region in cathepsin B. PMID- 9738466 TI - Possible stage-specific function of NF-kappaB during pre-B cell differentiation. AB - Lipopolysaccharide (LPS)-induced differentiation of the murine pre-B cell line 70Z/3 is a model for pre-B to B cell differentiation and has been used to show that the transcription factor NF-kappaB is essential to induce the expression of the Ig kappa gene. We have investigated the mechanism involved in late stages of the process when all cells have reached a more mature B phenotype, i.e. beyond 48 up to 96 h of LPS treatment. NF-kappaB binding activity was induced at early times by LPS treatment, but its DNA binding activity disappeared after 84 h of LPS treatment. Accumulation of IkappaB alpha protein in the nucleus correlated with the disappearance of NF-kappaB activity at 72, 84 and 96 h, and treatment of nuclear extracts of 72-96 h LPS-treated cells with Na-deoxycholate restored NF kappaB binding activity. The data indicate that NF-kappaB, while important to initiate the process of Ig kappa gene transcription in 70Z/3 pre-B cells, is no longer required for its maintenance in differentiated 70Z/3 cells. PMID- 9738467 TI - Identification of arginine-700 as the residue that binds the C-5 carboxyl group of 2-oxoglutarate in human lysyl hydroxylase 1. AB - Lysyl hydroxylase catalyzes the formation of hydroxylysine in collagens by a reaction that involves oxidative decarboxylation of 2-oxoglutarate. Its binding site can be divided into two main subsites: subsite I consists of a positively charged side-chain which binds the C-5 carboxyl group, while subsite II consists of two coordination sites of the enzyme-bound Fe2+ and is chelated by the C-1-C-2 moiety. In order to identify subsite I, we converted Arg-697, Arg-700 and Ser-705 individually to alanine and Arg-700 also to lysine, and expressed the mutant enzymes in insect cells. Arg-700-Ala inactivated lysyl hydroxylase completely, whereas Arg-697-Ala and Ser-723-Ala had only a relatively minor effect. Arg-700 Lys produced 93% inactivation under standard assay conditions, the main effect being a 10-fold increase in the Km for 2-oxoglutarate, whereas the Vmax was unchanged. Arg-700 thus provides the positively charged residue that binds the C 5 carboxyl group of 2-oxoglutarate, whereas Ser-705 appears to be of no functional significance in this binding. PMID- 9738468 TI - Retracing the evolution of amino acid specificity in glutaminyl-tRNA synthetase. AB - Molecular phylogenetic studies of glutaminyl-tRNA synthetase suggest that it has relatively recently evolved from the closely related enzyme glutamyl-tRNA synthetase. We have now attempted to retrace one of the key steps in this process by selecting glutaminyl-tRNA synthetase mutants displaying enhanced glutamic acid recognition. Mutagenesis of two residues proximal to the active site, Phe-90 and Tyr-240, was found to improve glutamic acid recognition 3-5-fold in vitro and resulted in the misacylation of tRNA(Gln) with glutamic acid. In vivo expression of the genes encoding these misacylating variants of glutaminyl-tRNA synthetase reduced cellular growth rates by 40%, probably as a result of an increase in translational error rates. These results provide the first biochemical evidence that glutaminyl-tRNA synthetase originated through duplication and consequent diversification of an ancestral glutamyl-tRNA synthetase-encoding gene. PMID- 9738469 TI - Biosynthetic processing and quaternary interactions of proprotein convertase SPC4 (PACE4). AB - SPC4 (PACE4), a member of the eukaryotic family of subtilisin-like proprotein convertases, is synthesized as a proenzyme (proSPC4) which undergoes proteolytic removal of N-terminal propeptide during transit through the secretory pathway. As this propeptide processing seems to be a key event in the functional expression of SPC4, we have investigated its mechanism and the intracellular site where it occurs. In transfected fibroblast cells, the 110-kDa proSPC4 undergoes slow cleavage to generate a 103-kDa mature enzyme in the endoplasmic reticulum (ER). Site-directed mutagenesis studies demonstrate that the proteolytic activation of SPC4 occurs mainly through a unimolecular autocatalytic process and propeptide cleavage is a prerequisite for its export from the ER. Sedimentation velocity and chemical cross-linking analysis demonstrate that the precursor protein in the cells exists as both a monomer and a dimer-sized complex whereas mature SPC4 exists only as a monomer. These results suggest that the cleavage of the N terminal propeptide of SPC4 plays a regulatory role in its activation and secretion through the change in its oligomeric state. PMID- 9738470 TI - Inhibitory effect of acidic pH on OmpC porin: wild-type and mutant studies. AB - By use of the patch clamp technique, we have compared the electrophysiological signature of OmpC porin channels at neutral and acidic pH. The perfusion of pH 5.4 buffer to the periplasmic side of excised patches promoted the closure or block of approximately 20% of the open porins present in the patch without changes in their single channel conductance. Besides this effect on the main, long-lived open state, lowering the pH also suppressed the spontaneous transitions of channels to another distinct short-lived open state. The inhibitory effect on the opening kinetics was particularly visible in two mutants (K16Q and E109Q) in which transitions to the short-lived open state are enhanced by the mutations themselves at pH 7.2. On the other hand, the R124Q mutant responded to acidic pH by an increased gating to the short-lived open state. The results suggest that acidic pH stabilizes a closed state of OmpC porin, and that the pH sensitivity might be conferred in part by R124, but not by K16 or E109. PMID- 9738471 TI - Analysis of deleted variant of hepatocyte growth factor by alanine scanning mutagenesis: identification of residues essential for its biological function and generation of mutants with enhanced mitogenic activity on rat hepatocytes. AB - To understand the structure-function relationship of hepatocyte growth factor (HGF) in more detail, we analyzed one of the other forms of HGF, deleted variant of HGF (dHGF), by alanine scanning mutagenesis. We show here that there are at least four sites important for dHGF to stimulate DNA synthesis in cultured adult rat hepatocytes, and that the residues of HGF essential for exerting its biological activity are not identical to those of dHGF. In addition, two mutants showed a decrease (approximately three-fold) in EC50 compared with wild-type dHGF in an assay of mitogenic activity on rat hepatocytes. PMID- 9738473 TI - Cloning of a human ether-a-go-go potassium channel expressed in myoblasts at the onset of fusion. AB - An early sign of human myoblast commitment to fusion is the expression of a non inactivating delayed rectifier K+ current, I(K(NI)), and an associated membrane potential hyperpolarization. We have isolated the full-length coding region of a human ether-a-go-go K+ channel (h-eag) from myoblasts undergoing differentiation. The h-eag gene was localized to chromosome 1q32-41, and is expressed as a approximately 9 kb transcript in myogenic cells and in adult brain tissue. Forced expression of h-eag in undifferentiated myoblasts generates a current with remarkable similarity to I(K(NI)) indicating that h-eag constitutes the channel responsible for this current in vivo. PMID- 9738472 TI - Cloning and characterization of a novel human inwardly rectifying potassium channel predominantly expressed in small intestine. AB - A new member of the two transmembrane domain potassium (K+) channel family was identified and isolated from a human brain cDNA library. The cDNA clone contains an open reading frame which encodes a 360 amino acid sequence with a characteristic P domain flanked by two hydrophobic regions representing the membrane spanning segments. The closest homologue of this gene product is the inwardly rectifying potassium channel subunit, Kir1.2 (identity approximately 42%). Northern blot analysis of human tissues with a selective cDNA probe for this new K+ subunit showed a single major transcript of 3.4 kb predominantly expressed at high levels in small intestine, with lower levels in stomach, kidney and brain. The main regions of expression in the central nervous system were medulla, hippocampus and corpus callosum. cRNA-injected oocytes and transiently transfected HEK293 cells expressed a K+ conductance which displays an inward rectification. This conductance is blocked by cesium and barium but is insensitive to tolbutamide and diazoxide even upon co-transfection of this novel subunit with the plasmid encoding the sulfonylurea receptor SUR1. Taken together, these results demonstrate that we have isolated and characterized a novel K+ channel subunit belonging to the inwardly rectifying K+ (Kir) channel family to which, upon homology classification, we have given the nomenclature Kir7.1. PMID- 9738474 TI - Density- and proliferation status-dependent expression of T-cadherin, a novel lipoprotein-binding glycoprotein: a function in negative regulation of smooth muscle cell growth? AB - The atypical low density lipoprotein (LDL) binding proteins (Mr 105 and 130 kDa; p105 and p130) in human aortic medial membranes and cultured human and rat aortic smooth muscle cells (SMC) have recently been identified as the cell adhesion glycoprotein T-cadherin. Although cadherins are generally recognized to be important regulators of morphogenesis, the function of T-cadherin in the vasculature is poorly understood. This study has examined the relationship between expression of T-cadherin and the density and proliferation status of SMC. T-cadherin (p105 and p130) levels in SMC lysates were measured on Western blots using ligand-binding techniques. T-cadherin expression was dependent upon cell density, and maximal levels were achieved at confluency. T-cadherin levels were reversibly modulated by switching cultures between serum-free (upmodulation) and serum-containing (downmodulation) conditions. Platelet-derived growth factor (PDGF)-BB, epidermal growth factor (EGF) or insulin-like growth factor (IGF) elicited a dose- and time-dependent downmodulation that was reversible after transfer of SMC to growth factor-free medium. Our results support the hypothesis that T-cadherin may function as a negative determinant of cell growth. PMID- 9738475 TI - The lack of extracellular Na+ exacerbates Ca2+-dependent damage of cultured cerebellar granule cells. AB - Rhodamine 123 staining, light and electron microscopy were used to evaluate the ultrastructural and functional state of cultured cerebellar granule cells after short treatment with the solution where NaCl was substituted by sucrose (sucrose balance salt medium, SBSM). Cell exposure to SBSM for 20 min resulted in the fact that mitochondria in the neurons lost their ability to sequester rhodamine 123. This effect could be prevented by: (i) non-competitive N-methyl-D-aspartate (NMDA) receptor channel blocker, 10(-5) M MK-801; (ii) a competitive specific antagonist of NMDA glutamate receptors, 0.25 x 10(-3) M D,L-2-amino-7 phosphonoheptanoate (APH); (iii) 10(-3) M cobalt chloride; (iv) removal of Ca2+ from the medium. Low Na+ in the Ca2+-containing medium caused considerable mitochondrial swelling in granule cells. However, the same treatment in the absence of calcium ions in the medium abolished the deleterious effect of SBSM on the neuronal mitochondrial structure and functions. It is suggested that (i) the exposure of cultured cerebellar granule cells to SBSM leads to a release of endogenous glutamate from cells; (ii) Ca2+ ions potentially de-energizing neuronal mitochondria enter the neuron preferentially through the NMDA channels rather than through the Na+/Ca2+ exchanger; (iii) mitochondrial swelling in granule cells is highly Ca2+-dependent; (iv) cellular overload with sodium ions can activate mitochondrial Na+/Ca2+ exchanger and thus prevent permeability transition pore opening in mitochondria. PMID- 9738476 TI - Inorganic anions induce state changes in spinach thylakoid membranes. AB - The role of cations in excitation energy distribution between the two photosystems of photosynthesis is well established. This paper provides evidence, for the first time, for an important role of anions in the regulation of distribution of absorbed light energy between the two photosystems. Inorganic anions caused redistribution of energy more in favour of photosystem I, as judged from measurements of chlorophyll a fluorescence transients, rates of electron transport in low light and 77 K fluorescence emission spectra: the Fv/Fm ratio was decreased by inorganic anions even in the presence of DCMU, the PS II electron transport was decreased whereas PS I electron transport was increased and the F735 (77 K emission from PS I)/F685 (77 K emission from PS II) ratio was increased. Such changes were observed with inorganic anions having different valencies (Cl- , SO4(2-), PO4(3-)): the higher the valency of the inorganic anion, the more the energy transferred towards PS I. Change in the valency of the inorganic anions thus regulates distribution of absorbed light energy between the two photosystems. However, organic anions like acetate, succinate, and citrate caused no significant changes in the Fv/Fm ratio, and in rates of PS I and PS II electron transport, showing their ineffectiveness in regulating light energy distribution. PMID- 9738477 TI - Photovoltage evidence that Glu-204 is the intermediate proton donor rather than the terminal proton release group in bacteriorhodopsin. AB - Electrogenic events in the E204Q bacteriorhodopsin mutant have been studied. A two-fold decrease in the magnitude of microsecond photovoltage generation coupled to M intermediate formation in the E204Q mutant is shown. This means that deprotonation of E204 is an electrogenic process and its electrogenicity is comparable to that of the proton transfer from the Schiff base to D85. pH dependence of the electrogenicity of M intermediate formation in the wild-type bacteriorhodopsin reveals only one component corresponding to the protonation of D85 in the bacteriorhodopsin ground state and transition of the purple neutral form into the blue acid form. Thus, the pK of E204 in the M state is close to the pK of D85 in the bacteriorhodopsin ground state (< 3) and far below the pK of the terminal proton release group (approximately 6). It is concluded that E204 functions as the intermediate proton donor rather than the terminal proton release group in the bacteriorhodopsin proton pump. PMID- 9738478 TI - Superoxide radical production by sponges Sycon sp. AB - Using the catechol Tiron as an O2-. scavenger, we showed that sea sponges (Sycon sp.) produce superoxide radicals in sea water at a high rate without any stimuli added. The rate of O2-. outflow from sponges to their water surroundings reaches a value of 0.5 nmol/min per sponge at pH 6.5. The generation of O2-. was inhibited by Cu,Zn-superoxide dismutase, and restored by the addition of KCN. We also confirmed the abiotic production of O2-. in sea water, detected earlier with a different method by Petasne and Zika [Nature 325 (1987) 516-518]. PMID- 9738479 TI - An intact conformation at the tip of elongation factor G domain IV is functionally important. AB - Three variants of Thermus thermophilus EF-G with mutations in the loop at the distal end of its domain IV were obtained. The replacement of His-573 by Ala and double mutation H573A/D576A did not influence the functional activity of EF-G. On the other hand, the insertion of six amino acids into the loop between residues Asp-576 and Ser-577 reduced the translocational activity of EF-G markedly, while its GTPase activity was not affected. It is concluded that the native conformation of the loop is important for the factor-promoted translocation in the ribosome. The functional importance of the entire EF-G domain IV is discussed. PMID- 9738480 TI - Sodium butyrate suppresses apoptosis in human Burkitt lymphomas and murine plasmacytomas bearing c-myc translocations. AB - We report that sodium butyrate, a natural product of fiber degradation by colonic bacteria, markedly suppresses c-Myc-mediated apoptosis induced in murine plasmacytomas and human Burkitt lymphomas by growth factor deprivation, but not in cell lines devoid of c-myc translocations. Attenuation of cell death is associated with downregulation of the rearranged c-myc and activation of pRb via its dephosphorylation. We suggest that in vivo sodium butyrate may play an important role in plasmacytomagenesis by supporting the survival of cells with c myc translocations, which otherwise would be eliminated by the lack of growth factors. PMID- 9738481 TI - Characterisation of high Mr wheat glutenin polymers by agarose gel electrophoresis and dynamic light scattering. AB - Agarose gel electrophoresis has been used to separate the complex mixture of wheat gluten polymers into fractions ranging in Mr, determined by dynamic light scattering, from about 500,000 to over 5x10(6). The separation is reliable and reproducible and well suited to the routine analysis of multiple samples. PMID- 9738482 TI - Advisers between a rock and a hard place. PMID- 9738483 TI - US drive to speed trials for rare disorders. PMID- 9738484 TI - Call for UK genetics food watchdog. PMID- 9738486 TI - Doubts over ability to monitor risks of BSE spread to sheep. PMID- 9738485 TI - China brings in regulations to put a stop to 'genetic piracy'. PMID- 9738487 TI - Too intelligent for our own good. PMID- 9738488 TI - Too intelligent for our own good. PMID- 9738490 TI - Evolutionary biology. Debatable homologies. PMID- 9738489 TI - Cancer. Has the smart bomb been defused? PMID- 9738491 TI - Neurobiology. Columns, slabs and pinwheels. PMID- 9738492 TI - RNA processing. A tale of two tails. PMID- 9738493 TI - Promiscuity in transgenic plants. PMID- 9738494 TI - Turning off follicular dendritic cells. PMID- 9738495 TI - Nerve agents degraded by enzymatic foams. PMID- 9738496 TI - Nematode phylogeny and embryology. PMID- 9738497 TI - A critical window for cooperation and competition among developing retinotectal synapses. AB - In the developing frog visual system, topographic refinement of the retinotectal projection depends on electrical activity. In vivo whole-cell recording from developing Xenopus tectal neurons shows that convergent retinotectal synapses undergo activity-dependent cooperation and competition following correlated pre- and postsynaptic spiking within a narrow time window. Synaptic inputs activated repetitively within 20 ms before spiking of the tectal neuron become potentiated, whereas subthreshold inputs activated within 20 ms after spiking become depressed. Thus both the initial synaptic strength and the temporal order of activation are critical for heterosynaptic interactions among convergent synaptic inputs during activity-dependent refinement of developing neural networks. PMID- 9738498 TI - Microbiological evidence for Fe(III) reduction on early Earth. AB - It is generally considered that sulphur reduction was one of the earliest forms of microbial respiration, because the known microorganisms that are most closely related to the last common ancestor of modern life are primarily anaerobic, sulphur-reducing hyperthermophiles. However, geochemical evidence indicates that Fe(III) is more likely than sulphur to have been the first external electron acceptor of global significance in microbial metabolism. Here we show that Archaea and Bacteria that are most closely related to the last common ancestor can reduce Fe(III) to Fe(II) and conserve energy to support growth from this respiration. Surprisingly, even Thermotoga maritima, previously considered to have only a fermentative metabolism, could grow as a respiratory organism when Fe(III) was provided as an electron acceptor. These results provide microbiological evidence that Fe(III) reduction could have been an important process on early Earth and suggest that microorganisms might contribute to Fe(III) reduction in modern hot biospheres. Furthermore, our discovery that hyperthermophiles that had previously been thought to require sulphur for cultivation can instead be grown without the production of toxic and corrosive sulphide, should aid biochemical investigations of these poorly understood organisms. PMID- 9738499 TI - When temporal terms belie conceptual order. AB - We conceive of time as a sequential order of real-world events, one event following another from past to present to future. This conception colours the way we speak of time ("we look forward to the time") and, as we show here, the way we process written statements referring to the temporal order of events, in real time. Terms such as 'before' and 'after' give us the linguistic freedom to express a series of events (real or imaginary) in any order. However, sentences that present events out of chronological order require additional discourse-level computation. Here we examine how and when these computations are carried out by contrasting brain potentials across two sentence types that differ only in their initial word ('After' X, Y versus 'Before' X, Y). At sites on the left frontal scalp, the responses to 'before' and 'after' sentences diverge within 300 ms; the size of this difference increases over the course of the sentences and is correlated with individual working-memory spans. Thus, we show that there are immediate and lasting consequences for neural processing of the discourse implications of a single word on sentence comprehension. PMID- 9738500 TI - Spontaneous pinwheel annihilation during visual development. AB - Neurons in the visual cortex respond preferentially to edge-like stimuli of a particular orientation. It is a long-standing hypothesis that orientation selectivity arises during development through the activity-dependent refinement of cortical circuitry. Unambiguous evidence for such a process has, however, remained elusive. Here we argue that, if orientation preferences arise through activity-dependent refinement of initially unselective patterns of synaptic connections, this process should leave distinct signatures in the emerging spatial pattern of preferred orientations. Preferred orientations typically change smoothly and progressively across the cortex. This smooth progression is disrupted at the centres of so-called pinwheels, where neurons exhibiting the whole range of orientation preferences are located in close vicinity. Assuming that orientation selectivity develops through a set of rules that we do not specify, we demonstrate mathematically that the spatial density of pinwheels is rigidly constrained by basic symmetry principles. In particular, the spatial density of pinwheels, which emerge when orientation selectivity is first established, is larger than a model-independent minimal value. As a consequence, lower densities, if observed in adult animals, are predicted to develop through the motion and annihilation of pinwheel pairs. PMID- 9738501 TI - lin-14 regulates the timing of synaptic remodelling in Caenorhabditis elegans. AB - In the nematode Caenorhabditis elegans six GABAergic motor neurons, known as DDs, remodel their patterns of synaptic connectivity during larval development. DD remodelling involves a complete reversal of the direction of information flow within nerve processes without marked changes in process morphology. We used a marker localized in vivo to DD presynaptic zones to analyse how the timing of DD remodelling is controlled. In wild-type animals, DDs remodel their synaptic outputs within a 3-5-hour period at the end of the first larval stage. We show that the heterochronic gene lin-14, which controls the timing of stage-specific cell lineages, regulates the timing of DD synaptic output remodelling. In lin-14 loss-of-function mutants, DDs remodel precociously. The degree of precocious remodelling is correlated with the level of lin-14 activity. Expression of lin 14(+) in the DDs of lin-14-null mutants rescues the precocious remodelling, indicating that lin-14 can act cell-autonomously. Consistent with this hypothesis, LIN-14 protein levels decrease in the DDs before remodelling. Our observations reveal a role of heterochronic genes in non-dividing cells, and provide an example of cell-autonomous respecification of neuronal connectivity. PMID- 9738502 TI - Three-dimensional segregation of supramolecular activation clusters in T cells. AB - Activation of T cells by antigen-presenting cells (APCs) depends on the complex integration of signals that are delivered by multiple antigen receptors. Most receptor-proximal activation events in T cells were identified using multivalent anti-receptor antibodies, eliminating the need to use the more complex APCs. As the physiological membrane-associated ligands on the APC and the activating antibodies probably trigger the same biochemical pathways, it is unknown why the antibodies, even at saturating concentrations, fail to trigger some of the physiological T-cell responses. Here we study, at the level of the single cell, the responses of T cells to native ligands. We used a digital imaging system and analysed the three-dimensional distribution of receptors and intracellular proteins that cluster at the contacts between T cells and APCs during antigen specific interactions. Surprisingly, instead of showing uniform oligomerization, these proteins clustered into segregated three-dimensional domains within the cell contacts. The antigen-specific formation of these new, spatially segregated supramolecular activation clusters may generate appropriate physiological responses and may explain the high sensitivity of the T cells to antigen. PMID- 9738503 TI - Yeast G1 cyclins are unstable in G1 phase. AB - In most eukaryotes, commitment to cell division occurs in late G1 phase at an event called Start in the yeast Saccharomyces cerevisiae, and called the restriction point in mammalian cells. Start is triggered by the cyclin-dependent kinase Cdc28 and three rate-limiting activators, the G1 cyclins Cln1, Cln2 and Cln3. Cyclin accumulation in G1 is driven in part by the cell-cycle-regulated transcription of CLN1 and CLN2, which peaks at Start. CLN transcription is modulated by physiological signals that regulate G1 progression, but it is unclear whether Cln protein stability is cell-cycle-regulated. It has been suggested that once cells pass Start, Cln proteolysis is triggered by the mitotic cyclins Clb1, 2, 3 and 4. But here we show that G1 cyclins are unstable in G1 phase, and that Clb-Cdc28 activity is not needed fgr G1 cyclin turnover. Cln instability thus provides a means to couple Cln-Cdc28 activity to transcriptional regulation and protein synthetic rate in pre-Start G1 cells. PMID- 9738504 TI - DNA hypomethylation leads to elevated mutation rates. AB - Genome-wide demethylation has been suggested to be a step in carcinogenesis. Evidence for this notion comes from the frequently observed global DNA hypomethylation in tumour cells, and from a recent study suggesting that defects in DNA methylation might contribute to the genomic instability of some colorectal tumour cell lines. DNA hypomethylation has also been associated with abnormal chromosomal structures, as observed in cells from patients with ICF (Immunodeficiency, Centromeric instability and Facial abnormalities) syndrome and in cells treated with the demethylating agent 5-azadeoxycytidine. Here we report that murine embryonic stem cells nullizygous for the major DNA methyltransferase (Dnmt1) gene exhibited significantly elevated mutation rates at both the endogenous hypoxanthine phosphoribosyltransferase (Hprt) gene and an integrated viral thymidine kinase (tk) transgene. Gene deletions were the predominant mutations at both loci. The major cause of the observed tk deletions was either mitotic recombination or chromosomal loss accompanied by duplication of the remaining chromosome. Our results imply an important role for mammalian DNA methylation in maintaining genome stability. PMID- 9738505 TI - RNA polymerase II is an essential mRNA polyadenylation factor. AB - Production of messenger RNA in eukaryotic cells is a complex, multistep process. mRNA polyadenylation, or 3' processing, requires several protein factors, including cleavage/polyadenylation-specificity factor (CPSF), cleavage stimulation factor, two cleavage factors and poly(A) polymerase. These proteins seem to be unnecessary for other steps in mRNA synthesis such as transcription and splicing, and factors required for these processes were not considered to be essential for polyadenylation. Nonetheless, these reactions may be linked so that they are effectively coordinated in vivo. For example, the CTD carboxy-terminal domain of the largest subunit of RNA polymerase II (RNAP II) is required for efficient splicing and polyadenylation in vivo, and CPSF is brought to a promoter by the transcription factor TFIID and transferred to RNAP II at the time of transcription initiation. These findings suggest that polyadenylation factors can be recruited to an RNA 3'-processing signal by RNAP II, where they dissociate from the polymerase and initiate polyadenylation. Here we present results that extend this model by showing that RNAP II is actually required, in the absence of transcription, for 3' processing in vitro. PMID- 9738506 TI - Identification of a membrane receptor for 1,25-dihydroxyvitamin D3 which mediates rapid activation of protein kinase C. AB - This paper is the first definitive report demonstrating a unique membrane receptor for 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) which mediates the rapid and nongenomic regulation of protein kinase C (PKC). Previous studies have shown that 1,25(OH)2D3 exerts rapid effects on chondrocyte membranes which are cell maturation-specific, do not require new gene expression, and do not appear to act via the traditional vitamin D receptor. We used antiserum generated to a [3H]1,25(OH)2D3 binding protein isolated from the basal lateral membrane of chick intestinal epithelium (Ab99) to determine if rat costochondral resting zone (RC) or growth zone (GC) cartilage cells contain a similar protein and if cell maturation-dependent differences exist. Immunohistochemistry demonstrated that both RC and GC cells express the protein, but levels are highest in GC. The binding protein is present in both plasma membranes and matrix vesicles and has a molecular weight of 66,000 Da. The 66 kDa protein in GC matrix vesicles has a Kd of 17.2 fmol/ml and Bmax of 124 fmol/mg of protein for [3H]1,25(OH)2D3. In contrast, the 66 kDa protein in RC matrix vesicles has a Kd of 27.7 fmol/ml and a Bmax of 100 fmol/mg of protein. Ab99 blocks the 1,25(OH)2D3-dependent increase in PKC activity in GC chondrocytes, indicating that the 1,25(OH)2D3-binding protein is indeed a receptor, linking ligand recognition to biologic function. PMID- 9738507 TI - Receptors for 1alpha,25(OH)2D3: past, present, and future. PMID- 9738508 TI - Reduced expression of interleukin-11 in bone marrow stromal cells of senescence accelerated mice (SAMP6): relationship to osteopenia with enhanced adipogenesis. AB - Aging is associated with an increase in bone marrow adipose tissue and a reduction in bone turnover. The P6 strain of senescence-accelerated mice (SAM) exhibit an early decrease in bone mass with a reduction in bone remodeling. In the bone marrow, suppressed osteoblastogenesis and osteoclastogenesis with enhanced adipogenesis are observed. The present study was undertaken to clarify the mechanism of age-related changes in bone turnover using bone marrow cells from SAMP6 mice. Because interleukin (IL)-11 has been shown to potently inhibit adipogenesis and to stimulate osteoclast formation, the effect of IL-11 on the differentiation of bone marrow cells was examined. The impaired formation of both osteoblasts and osteoclasts was restored and the enhanced formation of adipocytes was suppressed by the addition of 10 pM recombinant human IL-11. Other cytokines that activate gp130 as a common signal transducer, IL-6 and leukemia inhibitory factor, did not have such effects. Sequence analysis of the entire coding region of IL-11 cDNA obtained from SAMP6 stromal cells revealed no mutations. Constitutively secreted IL-11 protein into culture media, and its mRNA expression stimulated by transforming growth factor beta were reduced in stromal cells from SAMP6 compared with those in control mice. These results demonstrate that the expression of IL-11 is reduced in bone marrow cells of SAMP6 and suggest that the reduction in IL-11 actions is involved in the impairment of both osteoblastogenesis and osteoclastogenesis in these mice. There is a possibility that alterations in IL-11 actions may be associated with the age-related impairment in bone metabolism. PMID- 9738510 TI - An Sp1 binding site polymorphism in the COLIA1 gene predicts osteoporotic fractures in both men and women. AB - Genetic factors play an important role in the pathogenesis of osteoporosis, and recent studies have shown that a polymorphic Sp1 binding site in collagen type I alpha1 (COLIA1) gene is associated with bone mass and vertebral fractures in women from the U.K. Information on the predictive value of the COLIA1 Sp1 polymorphism in other populations is limited, however, and no studies have yet been performed in osteoporotic males. In view of this, we analyzed COLIA1 genotypes in relation to bone density and biochemical markers of bone turnover and the presence of osteoporotic fractures in a case-control study of Danish men and women. COLIA1 genotype was determined by polymerase chain reaction analysis of genomic DNA extracted from peripheral blood samples and related to bone mass, biochemical markers of bone turnover, and the presence of fracture in a study of 375 osteoporotic vertebral fracture patients and normal controls. There was no significant effect of COLIA1 genotype on bone mass or biochemical markers when data from the control group (n = 195) and fracture group (n = 180) were analyzed separately. However, the genotype distribution was significantly different in the fracture cases compared with age-matched controls (chi2 = 16.48, n = 249,p = 0.0003) due mainly to over-representation of the ss genotype in the fracture patients (14.3% vs. 1.4%), equivalent to an odds ratio for vertebral fracture of 11.83 (95% confidence interval 2.64-52.97) in those with the ss genotype. Similar differences in genotype distribution between osteoporotic patients and controls were observed in both men (chi2 = 11.52, n = 95, p = 0.0032, OR = 2.04) and women (chi2 = 6.90, n = 154, p = 0.032, OR = 1.37). In keeping with the above, logistic regression analysis showed that the ss genotype was an independent predictor of osteoporotic fracture (p = 0.028). This study confirms that the COLIA1 Sp1 polymorphism is significantly associated with osteoporotic vertebral fractures. The association is seen in both men and women, and the effect on fracture risk appears to be partly independent of bone mineral density. Our results raise the possibility that genotyping at the Sp1 site could be of clinical value in identifying individuals at risk of osteoporotic fractures in both genders. PMID- 9738509 TI - Effect of combination treatment with a vitamin D analog (OCT) and a bisphosphonate (AHPrBP) in a nude mouse model of cancer-associated hypercalcemia. AB - Hypercalcemia represents one of the important paraneoplastic syndromes affecting morbidity and mortality of cancer patients. We and others have demonstrated that vitamin D analogs with little calcemic activities suppress the transcription of the parathyroid hormone-related peptide (PTHrP) gene, a major humor responsible for cancer hypercalcemia, and thereby prevent the development of hypercalcemic syndrome. The present study was undertaken: to compare the therapeutic efficacy of a vitamin D analog, 22-oxa-1,25-dihydroxyvitamin D3 (OCT), and a bisphosphonate (disodium 3-amino-1-hydroxypropylidene-1,1-bisphosphonate pentahydrate [AHPrBP]), an inhibitor of osteoclastic bone resorption, on cancer induced hypercalcemia; and to see if the effect could be enhanced by combination treatment, using a nude mouse model implanted with a human pancreas carcinoma (FA 6). After a single intravenous administration, OCT (5 microg/kg of body weight [BW]) was as effective as AHPrBP (10 mg/kg of BW) in lowering blood ionized calcium levels in tumor-bearing nude mice, and their combination further enhanced the therapeutic effect. Although AHPrBP lost its efficacy after repeated injections, OCT was still effective after the third administration. The therapeutic effect of OCT in cancer hypercalcemia was observed in four other human tumors, including another pancreas carcinoma (PAN-7), two squamous cell carcinomas of the lung (KCC-C1 and LC-6), and a squamous carcinoma of the pharynx (PHA-1), all of which elaborated PTHrP into the circulation. Treatment with OCT resulted in a decrease in circulating PTHrP levels by approximately 50% in two representative models. However, the mechanism underlying the antihypercalcemic effect of OCT seemed complex, involving inhibition of PTHrP production, suppression of excessive bone resorption, and an antitumor activity. OCT also markedly inhibited the body weight loss with tumor growth, while AHPrBP, which exhibited a similar antihypercalcemic effect, was less effective than OCT in preventing cachexia. The anticachectic activity of their combination did not exceed that of OCT alone, suggesting a hypercalcemia-dependent as well as an independent mechanism of cancer cachexia. It is concluded that OCT may be useful, either as a single agent or in combination with bisphosphonates, for the treatment of cancer-associated hypercalcemia and cachexia. PMID- 9738511 TI - Extracellular calcium (Ca2+o)-sensing receptor in a mouse monocyte-macrophage cell line (J774): potential mediator of the actions of Ca2+o on the function of J774 cells. AB - The calcium-sensing receptor (CaR) is a G protein-coupled receptor that plays key roles in extracellular calcium ion (Ca2+o) homeostasis in parathyroid gland and kidney. Macrophage-like mononuclear cells appear at sites of osteoclastic bone resorption during bone remodeling and may play a role in the "reversal" phase following osteoclastic resorption and preceding bone formation. Bone resorption produces substantial local increases in Ca2+o that could provide a signal for bone marrow mononuclear cells in the vicinity, leading us to investigate whether such mononuclear cells express the CaR. In this study, we used the mouse J774 cell line, which exhibits a pure monocyte-macrophage phenotype. Both immunocytochemistry and Western blot analysis, using polyclonal antisera specific for the CaR, detected CaR protein in J774 cells. The use of reverse transcriptase polymerase chain reaction with CaR-specific primers, including a set of intron spanning primers, followed by nucleotide sequencing of the amplified products, also identified CaR transcripts in J774 cells. Exposure of J774 cells to high Ca2+o (2.8 mM or more) or the polycationic CaR agonist, neomycin (100 microM), stimulated both chemotaxis and DNA synthesis in J774 cells. Therefore, taken together, our data strongly suggest that the monocyte-macrophage cell line, J774, possesses both CaR protein and mRNA very similar, if not identical, to those in parathyroid and kidney. PMID- 9738512 TI - Thyroid hormone inhibits growth and stimulates terminal differentiation of epiphyseal growth plate chondrocytes. AB - As a continuation of our studies on mineralization in epiphyseal growth plate (GP) chondrocyte cultures, the effects of tri-iodothyronine (T3) in both beta glycerophosphate-containing, serum-free (HL-1) and beta-glycerophosphate-free, serum-containing medium (DATP5) were studied. The GP cells responded to T3 in a serum-, stage-, and dosage-dependent manner. Added at graded levels (0.1-10.0 nM) to preconfluent cultures (from day 7) in both HL-1 and DATP5, T3 caused progressive decreases in protein, collagen, and DNA synthesis but increased mineral deposition. In postconfluent cultures, these effects of T3 were generally muted. In preconfluent cultures, proteoglycan (PG) levels were not significantly affected in DATP5, although in HL-1 they were decreased by approximately 50%. In postconfluent cultures, T3 increased PG levels in DATP5 but had no effect in HL 1. In HL-1, alkaline phosphatase (ALP) activity was progressively increased by 200-500% in both pre- and postconfluent cultures. In DATP5 in preconfluent cultures, T3 initially stimulated but later suppressed ALP; in postconfluent cultures, T3 also transiently increased ALP but did not suppress activity upon longer exposure. The inhibitory effects of T3 on protein, PG, and DNA levels of GP chondrocytes suggest that in vivo its effects on bone growth must occur primarily after cellular proliferation. Apparently by binding to the 50 kDa thyroxine-binding globulin, which cannot penetrate the PG barrier, accessibility of T3 to GP chondrocytes is limited until the time of vascular penetration when its stimulatory effects on ALP and mineral deposition become critical for continued bone development. PMID- 9738513 TI - Calcitonin receptor binding properties in bone and kidney of the chicken during the oviposition cycle. AB - The binding property of calcitonin (CT) in the membrane fraction of calvaria and kidney of egg-laying and nonlaying hens was analyzed using a [125I] CT binding assay system. Binding properties of CT receptors in both tissues satisfy the authentic criteria of a receptor-ligand interaction in terms of specificity, reversibility, and saturation. Scatchard plots revealed a single class of binding sites. Values of the equilibrium dissociation constant (Kd) and binding capacity (Bmax) in laying hens showed a decrease during the period between 3 h before and 2 h after oviposition. No change was observed in nonlaying hens. In vivo administration of 17beta-estradiol or progesterone caused the decrease in Kd and Bmax values. The results suggest that the binding affinity and capacity of the CT receptor in the calvaria and the kidney of the hen may be modulated by the ovarian steroid hormone. PMID- 9738514 TI - Cysteine proteinases and matrix metalloproteinases play distinct roles in the subosteoclastic resorption zone. AB - Digestion of calvarial bone by osteoclasts depends on the activity of cysteine proteinases and matrix metalloproteinases (MMPs). It is unknown, however, whether these enzymes act simultaneously or in a certain (time) sequence. In the present study, this was investigated by culturing mouse calvarial bone explants for various time intervals in the presence or absence of selective low molecular weight inhibitors of cysteine proteinases (E-64, Z-Phe-Tyr(O-t-Bu)CHN2 or CA074[Me]) and MMPs (CI-1, CT1166, or RP59794). The explants were morphometrically analyzed at the electron microscopic level. All proteinase inhibitors induced large areas of nondigested demineralized bone matrix adjacent to the ruffled border of actively resorbing osteoclasts. The appearance of these areas proved to be time dependent. In the presence of the cysteine proteinase inhibitors, a maximal surface area of demineralized bone was seen between 4 and 8 h of culturing, whereas the metalloproteinase inhibitors had their maximal effect at a later time interval (between 16 and 24 h). Because different inhibitors of each of the two classes of proteolytic enzymes had the same effects, our data strongly suggest that cysteine proteinases attack the bone matrix prior to digestion by MMPs. In line with the view that a sequence may exist were differences in the amount of proteoglycans (shown with the selective dye cuprolinic blue) in the subosteoclastic demineralized areas induced by the inhibitors. In the presence of the cysteine proteinase inhibitor, relatively high levels of cuprolinic blue precipitates were found, whereas this was less following inhibition of metalloproteinases. These data suggested that cysteine proteinases are important for digestion of noncollagenous proteins. We propose the following sequence in the digestion of calvarial bone by osteoclasts: after attachment of the cell to the mineralized surface an area with a low pH is created which results in dissolution of the mineral, then cysteine proteinases, active at such a low pH, digest part of the bone matrix, and finally, when the pH has increased somewhat, MMPs exert their activity. PMID- 9738515 TI - Early changes in biochemical markers of bone turnover predict the long-term response to alendronate therapy in representative elderly women: a randomized clinical trial. AB - Although the antiresorptive agent alendronate has been shown to increase bone mineral density (BMD) at the hip and spine and decrease the incidence of osteoporotic fractures in older women, few data are available regarding early prediction of long-term response to therapy, particularly with regard to increases in hip BMD. Examining short-term changes in biochemical markers incorporates physiologic response with therapeutic compliance and should provide useful prognostic information for patients. The objective of this study was to examine whether early changes in biochemical markers of bone turnover predict long-term changes in hip BMD in elderly women. The study was a double-blind, placebo-controlled, randomized clinical trial which took place in a community based academic hospital. One hundred and twenty community-dwelling, ambulatory women 65 years of age and older participated in the study. Intervention consisted of alendronate versus placebo for 2.5 years. All patients received appropriate calcium and vitamin D supplementation. The principal outcome measures included BMD of the hip (total hip, femoral neck, trochanter, and intertrochanter), spine (posteroanterior [PA] and lateral), total body, and radius. Biochemical markers of bone resorption included urinary N-telopeptide cross-linked collagen type I and free deoxypyridinoline; markers of bone formation included serum osteocalcin and bone-specific alkaline phosphatase. Long-term alendronate therapy was associated with increased BMD at the total hip (4.0%), femoral neck (3.1%), trochanter (5.5%), intertrochanter (3.8%), PA spine (7.8%), lateral spine (10.6%), total body (2.2%), and one-third distal radius (1.3%) in elderly women (all p < 0.01). In the placebo group, bone density increased 1.9-2.1% at the spine (p < 0.05) and remained stable at all other sites. At 6 months, there were significant decreases in all markers of bone turnover (-10% to -53%, p < 0.01) in women on alendronate. The changes in urinary cross-linked collagen at 6 months correlated with long-term bone density changes at the hip (r = -0.35, p < 0.01), trochanter (r = -0.36, p < 0.01), PA spine (r = -0.41, p < 0.01), and total body (r = -0.34, p < 0.05). At 6 months, patients with the greatest drop in urinary cross-linked collagen (65% or more) demonstrated the greatest gains in total hip, trochanteric, and vertebral bone density (all p < 0.05). A 30% decrease in urinary cross-linked collagen at 6 months predicted a bone density increase of 2.8-4.1% for the hip regions and 5.8-6.9% for the spine views at the 2.5-year time point (p < 0.05). There were no substantive associations between changes in biochemical markers and bone density in the placebo group. Alendronate therapy was associated with significant long-term gains in BMD at all clinically relevant sites, including the hip, in elderly women. Moreover, these improvements were associated with early decreases in biochemical markers of bone turnover. Early dynamic decreases in urinary cross-linked collagen can be used to monitor and predict long-term response to bisphosphonate therapy in elderly women. Future studies are needed to determine if early assessment improves long-term patient compliance or uncovers poor compliance, thereby aiding the physician in maximizing the benefits of therapy. PMID- 9738516 TI - Prediction of vertebral and femoral strength in vitro by bone mineral density measured at different skeletal sites. AB - The aim of the present study was to investigate the prediction of vertebral and femoral strength in vitro by bone mineral density (BMD) measured at different skeletal sites. The third lumbar vertebral body, the right proximal femur, and the right calcaneus were removed from 38 male and 32 female cadavers (mean age 69 years, range 23-92 years). Areal BMD of all bone specimens was determined by dual energy X-ray absorptiometry (DXA). The failure load of the vertebral body and the femur was determined by mechanical testing. Vertebral and femoral strength were both greater in males than females (p < 0.01), as was BMD at all sites (p < 0.01). Vertebral strength correlated well with vertebral BMD (r2 = 0.64) but was only moderately correlated with BMD measured at the femur (r2 = 0.36) or the calcaneus (r2 = 0.18). Femoral strength showed the highest correlations with femoral BMD (r2 = 0.88) and somewhat weaker relationships with BMD at the vertebra (r2 = 0.50) and the calcaneus (r2 = 0.54). BMD values at the vertebra, femur, and calcaneus were only moderately interrelated (r2 = 0.31-0.65), and vertebral strength correlated only modestly with the strength of the femur (r2 = 0.36). These in vitro results support the concept that optimal prediction of vertebral or femoral strength by DXA requires site-specific assessments. PMID- 9738517 TI - The diuretic indapamide increases bone mass and decreases bone resorption in spontaneously hypertensive rats supplemented with sodium. AB - Clinical and epidemiological studies suggest that thiazide diuretics can prevent bone loss and decrease the incidence of hip fractures. However, the mechanism of the effect of diuretics on bone is not clearly established. Indapamide (IDP), a sulfonamide diuretic related to thiazides, is used to treat hypertension. Sixty spontaneously hypertensive rats (SHRs) were divided into four groups and treated with or without IDP (1.5 mg/kg/day) during 8 weeks in the presence or absence of a high sodium load (8% NaCl supplementation in the diet). Sodium and calcium excretions were increased in the rats receiving the high sodium load (SHR + 8% NaCl) comparatively with control rats (SHR). IDP decreased and increased, respectively, calcium and sodium excretions. Serum parathyroid hormone (PTH) was unchanged in any group. Bone density was measured at the femur, tibia, and vertebrae, and bone morphometry was performed at the metaphysis of the femur to evaluate bone architecture. Rats fed a high sodium diet had an average 5.5% decreased bone density at every site except the femoral diaphysis. The trabecular bone volume was also decreased (SHR + 8% NaCl vs. SHR, 11.99+/-0.78 vs. 17.51+/ 1.5%, p < 0.05). An increase in trabecular separation suggested that these changes were due to increased bone resorption. In the SHR + 8% NaCl + IDP group, IDP increased bone density and trabecular bone volume (SHR + 8% NaCl + IDP vs. SHR + 8% NaCl, 16.52+/-1.04 vs. 11.99+/-0.78%, p < 0.05). Trabecular separation and pyridinoline/creatinine excretion (SHR + 8% NaCl + IDP vs. SHR + 8% NaCl, 136.39+/-9.62 vs. 195.18+/-22.34 nmol/mmol, p < 0.05) were also decreased by IDP. These results show that in rats receiving a high sodium diet, IDP can reverse sodium-induced bone loss and increased bone resorption independently of changes in serum PTH. PMID- 9738518 TI - Variation in the short-term changes in bone cell activity in three regions of the distal femur immediately following ovariectomy. AB - The effect of ovariectomy (OVX) on cancellous bone in the rat is not uniform at all sites of the skeleton. We report variation in the short-term effects of adult OVX in three regions of the distal femur: the diaphysis (DIA), the metaphysis (META), and the epiphysis (EPI). Cancellous bone parameters were estimated in the three separate zones of the femora and compared with changes in bone cell activity, as estimated by osteoclast surface (Oc.S) and bone formation rate (BFR). Changes were studied for 30 days in a series of rats either sham-operated (Sham) or ovariectomized (OVX) at 7 months of age. Oc.S and BFR were elevated following OVX in all regions. The time course for the OVX-induced changes differed between regions: DIA, both Oc.S and BFR were elevated at day 9; META, Oc.S was also elevated at day 9, while the rise in BFR was delayed until day 21; EPI, Oc.S remained stable but increased relative to ovary-intact rats by day 18 due to reduced levels in the latter, but BFR did not rise until day 28. These changes in bone cell activity following OVX produced a 71% reduction of cancellous bone in the DIA and a 35% reduction in the META. In contrast, no OVX induced bone loss was observed in the EPI. This study shows that bone cell activity increases in each region of the distal femur within the first 30 days following OVX, independent of bone loss. However, the time course of increased bone cell activity is not uniform. These data highlight the role of local factors in the response to ovarian hormone deficiency. PMID- 9738519 TI - Bone loss, physical activity, and weight change in elderly women: the Dubbo Osteoporosis Epidemiology Study. AB - The present study examined the effects of physical activity, weight, and weight change on femoral bone loss in relation to age in elderly women. Baseline and follow-up measurements at an average interval of 2.7 years of femoral neck bone mineral density (BMD) were reanalyzed for 827 women who were part of the Dubbo Osteoporosis Epidemiology Study. Physical activity was assessed based on hours per day spent in each of various activities according to its expected oxygen consumption. The rate of loss of BMD progressively increased with age, i.e., 0.6+/-0.1, -1.1+/-0.2, and -2.1+/-0.6% per year (mean +/- SEM) for the 60-69, 70 79, and > or = 80 age groups, respectively (p < 0.001). After adjustment for age, physically inactive women lost more (-1.4+/-0.2% per year; p < 0.001), compared with physically active women (-0.5+/-0.3% per year, p = 0.15). Thinner women experienced more rapid bone loss, and women whose weight decreased (> or =5%) over the study period lost more bone (-1.7+/-0.4% per year) than those whose weight was stable (-0.8+/-0.1% per year) or increased (+0.1+/-0.3% per year, p < 0.01, analysis of variance). Furthermore, women whose BMD was high (>0.81 g/cm2) at baseline experienced greater loss (-1.1+/-0.2%) compared with those in the middle tertile (1.0+/-0.2%) or lowest tertile (-0.5+/-0.3%). Independent predictors of rate of bone loss included age, baseline BMD, weight, weight change, and physical activity; collectively these factors accounted for 13% of total variance of bone loss by multiple regression analysis. It is concluded that a physically active lifestyle and stable weight in the later decades of life may retard proximal femur bone loss and thus contribute to reduction of fracture risk. PMID- 9738520 TI - Falls among community-dwelling elderly in Japan. AB - Japanese have a lower incidence of hip fracture than Caucasians despite having lower bone mass. Hip fractures usually occur after a fall, and differing incidence rates of falls might explain the observed differences in hip fracture rates. To explore this hypothesis, we studied falls and related conditions among 1534 (624 men, 910 women) community-dwelling people aged 65 years and over in Japan and compared the prevalence of falls to Japanese-Americans living in Hawaii and to published studies of Caucasians. In Japan, 9% of the men and 19% of the women reported one or more falls during the past year. The prevalence of falls increased with age in both genders and was greater among women compared with men. In logistic regression models, having musculoskeletal disease, physical disability or limited activity increased the risk of falls by two to four times in both genders. Most fallers (92%) reported fear of future falls, and about one third of fallers reported that they went out less often as a result of their falls. Compared with native Japanese, the age-standardized prevalence of falls among Japanese-Americans was similar but about twice as high for Caucasians, which may explain the lower hip fracture risk of Japanese. PMID- 9738521 TI - Randomized controlled study of effects of sudden impact loading on rat femur. AB - Physical loading creating high peak strains on the skeleton at high strain rates is suggested to be the most effective type of activity in terms of bone mineral acquisition. This study assessed the effects of sudden impact loading on mineral and mechanical bone properties in 13-week-old Sprague-Dawley rats. The rats were randomly assigned as sedentary controls (SED, n = 10), control animals receiving low-intensity exercise (EX, n = 15), and experimental animals receiving low intensity exercise combined with sudden impact-loading (EX + IMP, n = 15). In the EX group, the rats walked in a walking mill at a speed of 10 cm/s for 20 minutes/day, 5 days/week for 9 weeks. In the EX + IMP group, the program was identical to the EX group except for the additional sudden impacts administered to their skeleton during the walking exercise. At the start, there were 50 impacts per session, after which their number was gradually increased to 200 impacts per session by week 6 and then kept constant until the end of the experiment, week 9. These horizontally and vertically directed body impacts were produced by a custom-made walking mill equipped with computer-controlled high pressure air cylinders. After sacrifice, both femora of each rat were removed and their dimensions, bone mineral content (BMC) by dual-energy X-ray absorptiometry, and mechanical properties by femoral shaft three-point bending and femoral neck compression were determined. The cortical wall thickness increased significantly in the EX and EX + IMP groups as compared with SEDs (+7.6%, p = 0.049 and +10%, p = 0.020, respectively). The EX + IMP group showed +9.0% (p = 0.046) higher cross sectional moment of inertia values than the EX group. No significant intergroup differences were seen in the BMC values, while the breaking load of the femoral shaft (EX + IMP vs. SED +8.8%,p = 0.047) and femoral neck (EX + IMP vs. SED +14.1%, p = 0.013) was significantly enhanced by the impact loading. In conclusion, this study indicates that mechanical loading can substantially improve the mechanical characteristics of a rat femur without simultaneous gain in its mineral mass. If this is true in humans too, our finding gives an interesting perspective to the numerous longitudinal exercise studies (of women) in which the exercise-induced gains in bone mass and density have remained mild to moderate only. PMID- 9738522 TI - Effect of recombinant human bone morphogenetic protein-2 on fracture healing in a goat tibial fracture model. AB - Bone morphogenetic proteins (BMPs) are considered to have important regulatory roles in skeletal embryogenesis and bone healing. Recombinant human BMPs (rhBMPs) have been shown to heal critical size defects and promote spinal fusion. We studied the effects of rhBMP-2 in an absorbable collagen sponge (ACS) on bone healing in a large animal tibial fracture model. Bilateral closed tibial fractures were created in 16 skeletally mature goats and reduced and stabilized using external fixation. In each animal, one tibia received the study device (0.86 mg of rhBMP-2/ACS or buffer/ACS), and the contralateral fracture served as control. The device was implanted as a folded onlay or wrapped circumferentially around the fracture. Six weeks following fracture, the animals were sacrificed and the tibiae harvested for torsional testing and histomorphologic evaluation. Radiographs indicated increased callus at 3 weeks in the rhBMP-2/ACS treated tibiae. At 6 weeks, the rhBMP-2/ACS wrapped fractures had superior radiographic healing scores compared with buffer groups and controls. The rhBMP-2/ACS produced a significant increase in torsional toughness (p = 0.02), and trends of increased torsional strength and stiffness (p = 0.09) compared with fracture controls. The device placed in a wrapped fashion around the fracture produced significantly tougher callus (p = 0.02) compared with the onlay application. Total callus new bone volume was significantly increased (p = 0.02) in the rhBMP-2/ACS fractures compared with buffer groups and controls regardless of the method of device application. The rhBMP-2/ACS did not alter the timing of onset of periosteal/endosteal callus formation compared with controls. Neither the mineral apposition rates nor bone formation rates were affected by rhBMP-2/ACS treatment. The increased callus volume associated with rhBMP-2 treatment produced only moderate increases in strength and stiffness. PMID- 9738523 TI - Paget's disease in a vertebral body: sixteen years of evolution in a young patient. PMID- 9738524 TI - Multigeneration exposure test of Drosophila melanogaster to ELF magnetic fields. AB - Mutations, other than dominant lethals, were accumulated on wild type second chromosomes (+) of Drosophila melanogaster during exposure to 50 Hz sinusoidal alternating magnetic fields of 0.5 or 5 mT (rms) for 40 generations by the Curly/Plum(Cy/Pm) accumulation method. We maintained, for 40 generations under continuous exposure, each (+) chromosome as a heterozygote with (Cy) chromosome. Viability of the (+) chromosome was tested by sib-mating of (Cy/+) male and (Cy/+) female in a culture every 10th generation to obtain the homozygote. Viability indices, defined as twice the ratio of number of (+/+) flies to that of (Cy/+) flies plus 1 in the progeny of the test mating, also were calculated, which equaled 1.00 at the starting point. For the control and 0.5 and 5 mT exposed groups, percent frequencies of recessive lethal lines, defined as a line with (+/+) flies less than 0.3% in the test mating, were, respectively, 1.9, 0.9, and 2.9% (10th), 9.0, 4.9, and 9.5% (20th), 30.3, 22.9, and 30.4% (30th), and 39.9, 32.4, and 43.3% (40th generation). For the control and 0.5 and 5 mT groups, average viability indices, excluding lethals and markedly deleterious, were, respectively, 0.778, 0.796, and 0.752 (20th), 0.704, 0.698, and 0.694 (30th), and 0.669, 0.678, and 0.595 (40th generation). Their decreasing rates were 0.0054, 0.0059, and 0.0078 per generation. No significant difference was detected among the exposure levels in either the recessive lethal mutation frequency or the viability index. PMID- 9738525 TI - Regional brain heating during microwave exposure (2.06 GHz), warm-water immersion, environmental heating and exercise. AB - Nonuniform heating may result from microwave (MW) irradiation of tissues and is therefore important to investigate in terms of health and safety issues. Hypothalamic (Thyp), cortical (Tctx), tympanic (Tty), and rectal (Tre) temperatures were measured in rats exposed in the far field, k-polarization (i.e., head pointed toward the transmitter horn and E-field in vertical direction) to two power densities of 2.06 GHz irradiation. The high-power density (HPM) was 1700 mW/cm2 [specific absorption rate (SAR): hypothalamus 1224 W/kg; cortex 493 W/kg]; the low-power density (LPM) was 170 mW/cm2 (SAR: hypothalamus 122.4 W/kg; cortex 49.3 W/kg). The increase (rate-of-rise, in degrees C/s) in Thyp was significantly greater than those in Tctx or Tre when rats were exposed to HPM. LPM produced more homogeneous heating. Quantitatively similar results were observed whether rats were implanted with probes in two brain sites or a single probe in one or the other of the two sites. The qualitative difference between regional brain heating was maintained during unrestrained exposure to HPM in the h-polarization (i.e., body parallel to magnetic field). To compare the temperature changes during MW irradiation with those produced by other modalities of heating, rats were immersed in warm water (44 degrees C, WWI); exposed to a warm ambient environment (50 degrees C, WSED); or exercised on a treadmill (17 m/min 8% grade) in a warm ambient environment (35 degrees C, WEX). WWI produced uniform heating in the regions measured. Similar rates-of-rise occurred among regions following WSED or WEX, thus maintaining the pre-existing gradient between Thyp and Tctx These data indicate that HPM produced a 2-2.5-fold difference in the rate-of-heating within brain regions that were separated by only a few millimeters. In contrast, more homogeneous heating was recorded during LPM or nonmicrowave modalities of heating. PMID- 9738526 TI - Temporal characteristics of transmission-line loadings in the Swedish childhood cancer study. AB - A recent study conducted in Sweden reported that 1) leukemia risk in children who lived near 220 or 400 kV electric-power transmission lines was associated with calculated historical magnetic field levels; 2) children living within a distance of 50 m of transmission lines had an elevated risk of leukemia; and 3) there was no association between leukemia and residential magnetic fields measured many years after diagnosis. Subsequently, these investigators found through logistic regression analysis that disease was more strongly associated with calculated historical fields than with distance. Since the calculated historical fields in that study depended predominantly on distance and transmission-line load current, the logistic regression results suggest that historical load current plays an important role in the epidemiological results. Thus, we studied hourly 1974 load current data for six transmission lines, and we examined 1958-1985 annual load current data for 112 transmission lines. Most lines exhibited marked diurnal load current rhythms during 1974, and all six showed systematic weekday-weekend differences. During 1958-1985, average loadings of Swedish 220 and 400 kV lines increased by about 1.3% year. Predictive-value and kappa-statistic analyses indicated that Swedish transmission-line load currents were not stable over long periods, so that contemporaneous load current (or a contemporary magnetic field measurement) was not a good surrogate for historical load current (or historical magnetic fields). The results provide a potential explanation of the failure of the Swedish Study to find an association between leukemia and contemporaneous magnetic field levels measured many years after the etiologic period, and suggest that the inclusion of load-current data could significantly improve the quality of historical field calculations. PMID- 9738527 TI - Electric field-induced changes in agonist-stimulated calcium fluxes of human HL 60 leukemia cells. AB - The mechanism of biological effects of extremely-low-frequency electric and magnetic fields may involve induced changes of Ca2+ transport through plasma membrane ion channels. In this study we investigated the effects of externally applied, low-intensity 60 Hz electric (E) fields (0.5 V/m, current density 0.8 A/m2) on the agonist-induced Ca2+ fluxes of HL-60 leukemia cells. The suspensions of HL-60 cells received E-field or sham exposure for 60 min and were simultaneously stimulated either by 1 microM ATP or by 100 microM histamine or were not stimulated at all. After E-field or sham exposure, the responses of the intracellular calcium levels of the cells to different concentrations of ATP (0.2 100 microM) were assessed. Compared with control cells, exposure of ATP-activated cells to an E-field resulted in a 20-30% decrease in the magnitude of [Ca2+]i elevation induced by a low concentration of ATP (<1 microM). In contrast, exposure of histamine-activated HL-60 cells resulted in a 20-40% increase of ATP induced elevation of [Ca2+]i. E-field exposure had no effect on non-activated cells. Kinetic analysis of concentration-response plots also showed that compared with control cells, exposure to the E-field resulted in increases of the Michaelis constant, Km, value in ATP-treated cells and of the maximal [Ca2+]i peak rise in histamine-treated HL-60 cells. The observed effects were reversible, indicating the absence of permanent structural damages induced by acute 60 min exposure to electric fields. These results demonstrate that low-intensity electric fields can alter calcium distribution in cells, most probably due to the effect on receptor-operated Ca2+ and/or ion channels. PMID- 9738529 TI - Effects of microwaves emitted by cellular phones on human slow brain potentials. AB - The influence of electromagnetic fields (EMF) emitted by cellular phones on preparatory slow brain potentials (SP) was studied in two different experimental tasks: In the first, healthy male human subjects had to perform simple self-paced finger movements to elicit a Bereitschaftspotential; in the second, they performed a complex and cognitive demanding visual monitoring task (VMT). Both tasks were performed with and without EMF exposure in counterbalanced order. Whereas subjects' performance did not differ between the EMF exposure conditions, SP parameters were influenced by EMF in the VMT: EMF exposure effected a significant decrease of SPs at central and temporo-parieto-occipital brain regions, but not at the frontal one. In the simple finger movement task, EMF did not affect the Bereitschaftspotential. PMID- 9738528 TI - Effects of exposure to static magnetic fields on the morphology and morphometry of mouse epididymal sperm. AB - Morphologic and morphometric sperm characteristics of mouse epididymal extracts from animals exposed to static magnetic fields were evaluated. For this purpose, animals were exposed for 35 days to a field of 0.7 T generated by a commercial permanent magnet for either 1 or 24 h per day. The values of morphometric parameters were obtained using the morphometric module of the Sperm Class Analyzer computerized image analysis system, and percentages of abnormalities were calculated. The size of sperm heads was unaffected by exposure to static magnetic fields. Lack of hook was a sperm head abnormality found significantly more frequently in animals exposed continually than in nonexposed animals, showing a possible alteration to the spermatogenic process after exposure to static magnetic fields. The percentage of sperm with coiled tails or of sperm with abnormal midpiece or tail was not altered by exposure. PMID- 9738530 TI - Epidermal ornithine decarboxylase and polyamines in mice exposed to 50 Hz magnetic fields and UV radiation. AB - We studied the influence of magnetic fields (MFs) and simulated solar radiation (SSR) on ornithine decarboxylase (ODC) and polyamines in mouse epidermis. Chronic exposure to combined MF and SSR did not cause persistent effects on ODC activity or polyamines compared to the animals exposed only to UV, although the same MF treatment was previously found to accelerate skin tumor development. In an acute 24-h experiment, an elevation of putrescine and down-regulation of ODC activity was observed in the animals exposed to a 100-microT MF. No effect was seen 24 h after a single 2-MED (minimal erythemal dose) exposure to SSR. The results indicate that acute exposure to 50 Hz MF does exert distinctive biological effects on epidermal polyamine synthesis. PMID- 9738531 TI - Behavioral satiety sequence (BSS) for the diagnosis of drug action on food intake. AB - The Behavioral Satiety Sequence (BSS) is the name given to the orderly transitions of eating, activity grooming and resting measured during the postingestive period. Because the BSS is considered to reflect the operations of natural physiological processes underlying satiety, the sequence can be used to discriminate between different drugs (and other manipulations) that reduce food intake via these natural physiological mechanisms or those that do so by interference. The BSS is only produced by the presence of a caloric load in the gut, and the preabsorptive satiety factors (such as CCK) the caloric load triggers. The BSS is most accurately defined by continuous observation rather than time or event sampling techniques [Partial Time Sampling (PTS) or Momentary Time Sampling (MTS)]. Continuous observation also allows the true duration and true frequency of each behavior to be analyzed. Continuous observation can be used to determine if the profiles associated with the reduction in food intake is caused by nausea, sedation, hyperactivity, or altered palatability of food. At the present time is it possible to identify a number of drugs whose suppression of food intake is associated with the disruption or preservation of the BSS. Drugs that increase synaptic 5-HT activity such d-fenfluramine, fluoxetine. and sibutramine all preserve the BSS and advance the onset of resting. The 5-HT1b/2c agonists mCPP and TFMPP and the 5-HT1b agonist CP-94,253 produce similar effects. However, the 5-HT2 agonist DOI and the 5-HT1a/1b agonist RU-24969 disrupt the BSS by inducing hyperactivity as does amphetamine. The 5-HT2 agonist MK-212 disrupts the BSS by inducing sedation. Selective dopamine agonists, at low doses, such as SKF-38393 (DA1) and LY-171555 (DA2) also preserve the BSS. However, detailed behavioral analysis of the effects of many recently discovered putative satiety factors remains to be carried out. PMID- 9738532 TI - (-)Ephedrine and caffeine mutually potentiate one another's amphetamine-like stimulus effects. AB - Using rats trained to discriminate 1 mg/kg of (+)amphetamine (ED50 = 0.4 mg/kg) from saline vehicle in a two-lever drug discrimination procedure, it was shown that (-)ephedrine (ED50 = 4.5 mg/kg), but not (+)ephedrine, substitutes for the (+)AMPH stimulus. It was also shown that caffeine (ED50 = 12.9 mg/kg) can substitute for (+)amphetamine in a dose-related fashion. Doses of (-)ephedrine and caffeine, which produced < or = 1% drug-appropriate responding when administered alone, were able to enhance each other's stimulus effects when administered in combination such that there was a twofold leftward shift in their respective dose-response curves. Furthermore, stimulus generalization occurred when a dose of caffeine that produced saline-appropriate responding when administered alone was administered in combination with (+)ephedrine. It would appear that low doses of (-)ephedrine and caffeine may mutually potentiate one another's stimulus effects in (+)AMPH-trained rats, and that a combination of caffeine and (+)ephedrine result in altered stimulus character when compared to comparable doses of either agent administered alone. PMID- 9738533 TI - No self-injurious behavior was found in HPRT-deficient mice treated with 9 ethyladenine. AB - It has been reported that 9-ethyladenine (9-EA) is an efficient inhibitor of APRT (adenine phosphoribosyltransferase) and that its administration causes self injurious behavior (Lesch-Nyhan Syndrome-like symptoms) in HPRT (hypoxanthine guanine phosphoribosyltransferase)-deficient mice. In contrast, we found neither any self-injurious behavior (SIB), such as visible injury or hair loss, nor any apparent decrease in APRT activity in HPRT-deficient mice treated with 9-EA. We also found that 9-EA has little irreversible or competitive inhibitory effect on APRT in vitro, even at a concentration of 10(-2) M. In light of the negative finding of SIB in APRT/HPRT double-deficient mice, it seems unlikely that SIB in HPRT-deficient mice is caused by lowered APRT activity. It is concluded that 9-EA is not a sufficient APRT inhibitor and cannot be used in experiments that mimic lowered APRT status in an animal model. PMID- 9738535 TI - Adrenergic and cholinergic inputs in preoptic area of rats interact for sleep wake thermoregulation. AB - Isolated studies have shown that both norepinephrine and acetylcholine into the medial preoptico-anterior hypothalamic area tonically regulate sleep-wake and body temperature. A possible interaction between these neurotransmitters for the regulation of such functions has been investigated in this study. To study this interaction a combination of either prazosin and carbachol or, scopolamine and methoxamine was injected into the medial preoptico-anterior hypothalamic area and the effect on sleep, wake, and rectal temperature recorded simultaneously. The combination of chemicals were selected based on our previous studies where it was observed that each of the chemicals in a combination had opposite effects. It was observed that injection of the combination expressed a resultant summated effects of individual component chemicals when injected in isolation (observed in previous studies). Because effect of neither of the chemicals in the combination was dominant, the results suggest an interaction and integration of the adrenergic and cholinergic inputs in the medial preoptico-anterior hypothalamic area for the regulation of sleep-wakefulness and body temperature. PMID- 9738534 TI - Responding for rewarding brain stimulation: cocaine and isradipine plus naltrexone. AB - Rats, fixed with chronically indwelling electrodes for electrical intracranial stimulation (ICS) of the lateral hypothalamus, were taught to press a bar for ICS. Once pressing rates became stable, during daily 20-min sessions, rats were given cocaine (5 or 20 mg/kg) before the sessions. When given daily, cocaine consistently enhanced rates of pressing. When a combination of small doses of isradipine (e.g., 1 mg/kg) and naltrexone (3 mg/kg) were given before cocaine administration. the combination blocked cocaine's enhancement of pressing for ICS. The combination, however, neither reduced rates of pressing below those observed under placebos (i.e., baseline conditions) nor reduced rates when no cocaine was given. Naltrexone and isradipine (in the dose used in the combination) by themselves did not block cocaine's effects. This profile of effects indicates that a combination of isradipine and naltrexone is apt to be useful in treating cocaine use disorders. PMID- 9738536 TI - Multiphasic consequences of the acute administration of ethanol on cerebral glucose metabolism in the rat. AB - The present study investigated the role of the postinjection interval in determining the functional consequences of acute ethanol administration in the CNS. Local cerebral metabolic rates for glucose (LCMRglc) were determined by the 2[14C]deoxyglucose method in 48 brain structures of ethanol-naive Sprague-Dawley rats. Tracer was injected 1 or 45 min after a 0.8 g/kg intragastric dose of ethanol or water. At the early time point, LCMRglc was increased in a highly restricted portion of the basal ganglia that included the dorsal striatum, globus pallidus, and core of the nucleus accumbens, compared to water controls. No significant decreases were found at this early time point. At the later time point, by contrast, LCMRglc was decreased in a different set of brain structures. These sites were limbic in nature and included the infralimbic and anterior cingulate cortices, dentate gyrus, lateral septum, and the bed nucleus of the stria terminalis. These data indicate that there are multiple phases that can be detected during the time course of an acute dose of ethanol. They further demonstrate the involvement of different neural systems at the two time points. Increased activity in basal ganglia is consistent with stimulated motor activity, whereas diminished activity in limbic sites may correspond to changes in mood and motivation. PMID- 9738537 TI - Avoidance of time-out from response-independent food presentation: effects of chlordiazepoxide and buspirone. AB - Five male Wistar rats were exposed to a two- component multiple schedule. In one component, signaled by a tone, food pellets were presented on a random-time 120-s schedule. In the other component, food pellets were presented on a random-time 30 s schedule. Pellets were only presented during a 10-s time-in period that alternated with a 50-s time-out period, unless the subject pressed a lever to postpone time-out presentation by 20 s. Response-independent food pellets were never presented within 2 s of this avoidance response. For most subjects avoidance rates were consistently higher when response-independent food pellets were delivered infrequently than when they were delivered more often. The amount of time spent in time-in varied considerably between subjects but was not consistently related to the frequency of response-independent pellet presentation. Once stable response rates were established subjects were intraperitoneally injected with different doses of chlordiazepoxide (1, 3, 10, 17, or 30 mg/kg) or buspirone (0.1, 0.3, 1.0, 1.7, 3.0, or 4.2 mg/kg). Low doses of chlordiazepoxide either did not affect or slightly increased avoidance response rates, whereas higher doses (10 mg/kg and up) produced a dose-dependent decrease in avoidance responding. The time subjects spent in the presence of stimuli associated with the availability of response-independent food either did not change or increased slightly after the lower doses of chlordiazepoxide, while it decreased dose dependently following the higher doses. Low doses of buspirone increased avoidance rates in subjects first exposed to chlordiazepoxide, but did not alter rates in the remaining subjects. Intermediate doses of buspirone decreased avoidance rates more in the component with the lower frequency of pellet presentation, higher doses further decreased response rates. The amount of time spent in the presence of stimuli associated with pellet presentation was minimally affected by the lower doses of buspirone, but decreased dose dependently following the higher doses. The results of this experiment add further credence to the notion that the behavioral effects of drug administration may depend on nonpharmacological variables including, but not limited to, the nature of the consequent event. PMID- 9738538 TI - A simple device for visualising pain during surgical procedures under monitored anesthesia care. PMID- 9738539 TI - Induction of apoptosis by laser: a new therapeutic modality. PMID- 9738540 TI - Collagen thermal damage and collagen synthesis after cutaneous laser resurfacing. AB - BACKGROUND AND OBJECTIVE: Objectively measure and compare postoperative collagen thermal damage and subsequent new collagen synthesis after cutaneous laser resurfacing using two carbon dioxide laser systems. STUDY DESIGN/MATERIALS AND METHODS: We created 240 resurfacing wounds on eight piglets with scanned and short-pulsed lasers using the manufacturer's suggested settings. The wounds varied with respect to the number of laser passes and postoperative survival times. Samples were harvested for histological analysis. RESULTS: The scanned laser resulted in an average of 52% more collagen thermal damage on the day of surgery (P < 0.0001) and an average of 78% more thermal damage 3 days postoperative (P < 0.0001) than the short-pulsed laser. The amount of new collagen synthesis correlated with the amount of thermal damage, with the scanned laser wounds showing 44% greater new collagen synthesis than the short-pulsed laser wounds on postoperative day 7 (P < 0.0001) and 48% greater new collagen synthesis on postoperative day 14 (P < 0.0001). CONCLUSION: Compared to the short pulsed laser, the scanned laser results in a greater depth of collagen thermal damage with a correspondingly greater depth of new collagen synthesis after cutaneous resurfacing. PMID- 9738541 TI - Leg telangiectasia treatment with a 1.5 ms pulsed dye laser, ice cube cooling of the skin and 595 vs 600 nm: preliminary results. AB - BACKGROUND AND OBJECTIVE: Preliminary results indicate that pulsed dye lasers (PDL) with 1.5 ms pulsewidth and 595 nm wavelength are effective in the treatment of leg telangiectasia. The aim of this study was to evaluate if the clinical results could be improved by a) an effective skin cooling with ice cubes and b) the longer wavelength of 600 nm. STUDY DESIGN/MATERIALS AND METHODS: In 87 patients with vessels up to 1 mm in diameter, 257 single test treatments were performed using wavelengths of 595 and 600 nm, fluences of 16, 18, and 20 J/cm2, a 1.5 ms pulse duration, and an elliptical spot of 2 x 7 mm. In 7 patients, the skin surface temperature curve was measured after cooling with ice cubes vs hydrogel dressings, and spot geometry and fluence were investigated with and without the gel dressing. RESULTS: Vessel clearance was evaluated 6-8 weeks after treatment. 20 J/cm2 were most effective (80% clearance >50%), and 18 J/cm2 were more effective than 16 J/cm2 (66.2 vs 52.5% clearance >50%). There was a tendency towards better results with 595 nm, but the differences were not significant. Vessels with a diameter <0.5 mm cleared significantly better than those with 0.5 1 mm (69.1 vs 31.9% clearance >50%). Hypo- and hyperpigmentation were seen in 32% of the patients. Cooling with ice cubes proved to be far more effective than with hydrogel dressings (temperature decrease approx 15 vs 5 degrees C). Additionally, the gel dressing caused an energy loss of approx 35% and an irregular spot geometry as shown on burn paper. CONCLUSIONS: Treatment of leg telangiectasia with the 1.5 ms-PDL is safe and effective, especially in vessels smaller than 0.5 mm in diameter; 595 nm and 18 J/cm2 seem to be somewhat more effective as 600 nm and 16 J/cm2; and 20 J/cm2 are even more effective, but persistent hyperpigmentation cannot yet be excluded due to insufficient follow-up time. Cooling with ice cubes is more effective and less expensive than gel dressings, and the short term clinical results are equivalent, even if the frequency of transient pigmentary changes is increased. PMID- 9738542 TI - Potential role of transoral laser surgery for larynx carcinoma. AB - BACKGROUND AND OBJECTIVE: The treatment of larynx carcinoma is not settled to date. This prospective study evaluates the potential role of transoral laser surgery (TLS) for larynx carcinoma in a large series of unselected patients from a single institution. MATERIALS AND METHODS: A total of 504 consecutive patients with previously untreated carcinoma of the larynx were seen from 1986-1994. Their treatment modalities and results were prospectively evaluated. RESULTS: TLS was used in 290 patients (58%), total laryngectomy in 130 (26%), conventional partial laryngectomies in 31 (6%), and radiotherapy in 34 (7%). Nineteen (4%) had no curative treatment. Uncorrected actuarial survival for all patients with glottic carcinoma stages I and II treated with laser surgery (n = 202) was 80.2%, cause specific survival 96.7%, and local control 85.8%. Uncorrected actuarial survival for all patients with supraglottic carcinoma stages I and II treated with laser surgery (n = 40) was 49.0%, cause specific survival 78.6%, and local control 87.3%. CONCLUSION: TLS was the most important single treatment modality in this large series of unselected patients. It is a safe and time- and cost-effective alternative to radiotherapy for early stage larynx carcinoma. PMID- 9738543 TI - Three-dimensional laser imaging system for measuring wound geometry. AB - BACKGROUND AND OBJECTIVE: A low cost laser imager was designed and fabricated for measurement of wound geometry. METHODS: The accuracy of the imager was validated using reference depressions of known dimensions. Perimeter, area, and volume were compared to planimetric and packing techniques on simulated wound models. RESULTS: Wound tracing and alginate measurement methods required approximately 20 times longer for the reference standards, and 11 times longer for the simulated wounds than with the laser scanning method (LSM). LSM consistently overestimated the reference perimeter by 0.73+/-0.20 cm and the area by 0.98+/-0.62 cm2. Volume estimates were not statistically different. The tracing method underestimated the perimeter by 0.34+/-0.27 cm and the area by 1.07+/-1.09 cm2. Volume measurements by the alginate method were not statistically different. The perimeters of the simulated wounds averaged 1.29+/-0.27 cm greater using the LSM than obtained by the tracing method, and areas greater by 2.02+/-1.30 cm2. Volume scans averaged 1.04+/-0.61 cm3 greater than by the alginate method. PMID- 9738544 TI - In situ temperature measurements with thermocouple probes during laser interstitial thermotherapy (LITT): quantification and correction of a measurement artifact. AB - BACKGROUND AND OBJECTIVE: The purpose of this work was to quantify the magnitude of an artifact induced by stainless steel thermocouple probes in temperature measurements made in situ during experimental laser interstitial thermo-therapy (LITT). A procedure for correction of this observational error is outlined. STUDY DESIGN/MATERIALS AND METHODS: A CW Nd:YAG laser system emitting 20W for 25-30 s delivered through a fiber-optic probe was used to create localized heating. The temperature field around the fiber-optic probe during laser irradiation was measured every 0.3 s in air, water, 0.4% intralipid solution, and fatty cadaver pig tissue, with a field of up to fifteen needle thermocouple probes. RESULTS: Direct absorption of Nd:YAG laser radiation by the thermocouple probes induced an overestimation of the temperature, ranging from 1.8 degrees C to 118.6 degrees C in air, 2.2 degrees C to 9.9 degrees C in water, 0.7 C to 4.7 C in intralipid and 0.3 C to 17.9 C in porcine tissue after irradiation at 20W for 30 s and depending on the thermocouple location. The artifact in porcine tissue was removed by applying exponential and linear fits to the measured temperature curves. CONCLUSION: Light absorption by thermocouple probes can induce a significant artifact in the measurement of laser-induced temperature increases. When the time constant of the thermocouple effect is much smaller than the thermal relaxation time of the surrounding tissue, the artifact can be accurately quantified. During LITT experiments where temperature differences of a few degrees are significant, the thermocouple artifact must be removed in order to be able accurately to predict the treatment outcome. PMID- 9738545 TI - Erbium: YAG laser trabecular ablation (LTA) in the surgical treatment of glaucoma. AB - BACKGROUND AND OBJECTIVE: Laser trabecular ablation (LTA) aims to remove trabecular tissue and to open Schlemm's canal in order to improve outflow facility in glaucomatous eyes. The purpose of our study was to investigate the pressure-reducing effect of LTA in chronic open-angle glaucoma 1 year following laser surgery. STUDY DESIGN/MATERIALS AND METHODS: Eleven eyes of 11 patients with chronic open-angle glaucoma were treated by circumscribed laser ablation of the trabecular meshwork. We used an Erbium:YAG laser (2.94 microm) with a quartz fiber contact endoprobe (320 microm core-diameter, 385 microm coating-diameter) applying 11-30 single neighbouring laser pulses (5-7 mJ) to the trabecular meshwork by an ab-interno approach. Laser procedure was gonioscopically visualized. RESULTS: Mean maximum intraocular pressure (IOP) of all 11 patients before LTA was 36 mmHg and dropped down to 22 mmHg after a mean follow-up of 12 months following LTA; this represents an IOP decrease of 38% (P=0.008). The average number of medications per eye dropped from 2.8 to 1.5 per eye (P=0.021). CONCLUSION: Although IOP lowering effect of Erbium:YAG laser trabecular ablation did not prove as effective as in filtering procedures, LTA might be a valuable alternative in glaucoma surgery especially in order to avoid conjunctival scarring and postoperative hypotony. PMID- 9738546 TI - Do leptin levels predict weight gain?--A 5-year follow-up study in Mauritius. Mauritius Non-communicable Disease Study Group. AB - OBJECTIVE: To investigate whether relative baseline leptin levels predict long term changes in adiposity and/or its distribution. RESEARCH METHODS AND PROCEDURES: In a longitudinal study of 2888 nondiabetic Mauritians aged 25 years to 74 years who participated in population-based surveys in 1987 and 1992, changes in body mass index (BMI), waist/hip ratio (WHR), and waist circumference were compared between "hyperleptinemic," "normoleptinemic," and "hypoleptinemic" groups. "Relative leptin levels" were calculated as standardized residuals from the regression of log10 leptin on baseline BMI to provide a leptin measure independent of BMI. Analyses were performed within each sex. A linear regression model was used to assess the effect of standardized residuals on changes in BMI, WHR, and waist circumference, independent of baseline BMI, age, fasting insulin, and ethnicity. RESULTS: After adjusting for age and baseline BMI by analysis of covariance, there was no difference in changes in BMI, WHR, or waist circumference between men with low, normal, or high relative leptin levels. Among women, there was a significant difference in deltaWHR across leptin groups, such that the largest increase occurred in the "normal" leptin group. For both men and women, the linear regression models explained approximately 10% of variation in dependent variables, and the only significant independent variables were age, BMI, and being of Chinese origin, compared with Indian origin. DISCUSSION: These findings do not support a role for leptin concentration in predicting weight gain or changes in fat distribution in adults over a 5-year period. PMID- 9738547 TI - Body composition changes in Caucasian and African American children and adolescents with obesity using dual-energy X-ray absorptiometry measurements after a 10-week weight loss program. AB - OBJECTIVE: Changes in body composition during a weight loss program have not been described in children. We wanted to test the hypothesis that weight loss can be achieved while maintaining total body fat-free mass. RESEARCH METHODS AND PROCEDURES: We determined body composition changes by using dual-energy X-ray absorptiometry measured at baseline and after the first 10 weeks of a multidisciplinary weight loss program. The program consisted of 10 weekly group sessions where the children were provided instruction in lifestyle modification, including diet and exercise. Program leaders included a pediatrician, psychologist, registered dietitian, and exercise instructor. RESULTS: We studied 59 obese children, mean (+/-SD) age 12.8+/-2.6 years, 29% boys and 71% girls, 49% Caucasian, and 51% African American. At enrollment, the children's mean height and body mass index were 157 cm and 38.9 kg/m2, respectively. The children's dual energy X-ray absorptiometry-derived mean at baseline and at 10 weeks and corresponding p values were: weight (94.6 kg vs. 92.3 kg, p<0.0001), total body fat mass (46.9 kg vs. 44.3 kg, p<0.0001), percentage total body fat (49.2% vs. 47.5%, p<0.0001), total trunk mass (43.0 kg vs. 41.5 kg, p<0.0001), total trunk fat (21.2 kg vs. 20.0 kg, p<0.0001), total body fat-free mass (47.6 kg vs. 47.9 kg, p=0.33), total body bone mass (2.7 kg vs. 2.7 kg, p=0.99), and total body bone mineral density (1.14 g/cm2 vs. 1.15 g/cm2, p=0.0119). The children's race, gender, or Tanner stage did not affect these changes. DISCUSSION: Decreases in total body fat mass was achieved, and total body fat-free mass was maintained among boy and girl Caucasian and African American children participating in this lifestyle modification weight loss program. PMID- 9738548 TI - Dieting, exercise, or disordered eating does not account for extremes of body weight within families. AB - OBJECTIVE: Families having both members with obesity and thin members should contain substantial information for genetics studies, provided measured phenotype is an accurate indicator of genetic predisposition. We assessed the impact of potentially complicating behavioral factors on obesity phenotypes of family members selected for a long-term project to identify genes for human obesity. RESEARCH METHODS AND PROCEDURES: Ninety-nine Caucasian families were selected for study because they contained both extremely obese and average-weight family members. Family members (n=492) were queried about their diet and exercise habits, their psychiatric histories as they pertained to eating disorders, and for a subset of subjects (n=329), a lifetime dieting history and a lifetime maximum weight were recorded. RESULTS: Subjects with average body weights in these families did not appear to be maintaining their weight by dieting and <4% of the average-weight subjects had ever been obese in the past. DISCUSSION: Although dieting and other weight loss practices potentially could either mask or complicate the genotype-phenotype relationship, we found little evidence for this possibility in the families studied. PMID- 9738549 TI - Psychiatric ill-health of women and its relationship to obesity and body fat distribution. AB - OBJECTIVE: Abdominal obesity is associated with serious, prevalent diseases. Previously, psychiatric symptoms and ill-health has been found in this condition in men. The results of a similar study in women is reported herein. RESEARCH METHODS AND PROCEDURES: A cohort of 1464 women, aged 40 years and recruited by systematic sampling, was examined (77.7% participation rate). Items regarding use of anxiolytics, hypnotics, and antidepressive drugs were registered, as well as symptoms of dyspepsia, sleeping disturbances, melancholy, and degree of life satisfaction. Smoking and alcohol consumption, as well as self-measured weight, height, waist, and hip circumferences, were reported, from which body mass index [BMI; weight (kg)/height2 (m2), kg/m2] and the waist/hip circumference ratio (WHR) were calculated. RESULTS: In bivariate analyses, BMI was associated with use of anxiolytics, antidepressive drugs, various sleeping disturbances, and a low degree of life satisfaction. After controlling for the WHR, alcohol, and tobacco use in multivariate analysis, the associations between BMI and use of anxiolytics and sleeping disturbances remained significant. The WHR correlated with dyspepsia, sleeping problems, and use of antidepressive drugs. After adjustments for BMI, smoking, and alcohol, the relationship to dyspepsia and antidepressants remained significant. DISCUSSION: The results suggest that elevated BMI (obesity) and elevated WHR (central fat distribution) are associated in different ways with symptoms of psychiatric ill-health in women. Obesity alone shows no such relationships to psychiatric ill-health in men, whereas central fat distribution shows independent associations to all of the measured variables studied in this report in women, suggesting gender differences in these associations. PMID- 9738550 TI - Clinic-based vs. home-based interventions for preventing weight gain in men. AB - OBJECTIVE: Weight gain occurs frequently in men aged 25-40. This study compared the effectiveness of a clinic-based and a home-based intervention with a no treatment control group in preventing this weight gain. RESEARCH METHODS AND PROCEDURES: Men (n=67)-aged 25 to 40, sedentary, with a body mass index of 22 to 30, recruited from the University of Pittsburgh-were randomly assigned to 4-month treatments focused on increasing aerobic exercise and reducing fat intake through a clinic-based (CB) or a home-based (HB) program, or to a delayed-treatment control group. Subjects were reassessed at 4 months. RESULTS: Adherence and outcome did not differ significantly between the CB and HB programs, except that CB subjects recorded their food intake more frequently, and a greater number of CB subjects achieved a total of 120 miles of exercise over the 4 months. Subjects in the two intervention conditions combined lost significantly more weight ( 1.6+/-2.5 kg) than control subjects, who gained 0.2+/-1.9 kg (p<0.01); this effect of treatment was seen primarily in men with a body mass index of 27 to 30 (-2.7 kg for CB and HB combined vs. + 1.5 kg for control). Treated subjects also had somewhat greater improvements in body composition, aerobic fitness, and weekly energy expenditure than controls, although these differences did not reach significance. DISCUSSIONS: Both CB and HB intervention show promise in preventing weight gain in young men, especially in those who are slightly overweight. Larger studies, using more representative samples of young men, appear warranted. PMID- 9738551 TI - Monkey leptin receptor mRNA: sequence, tissue distribution, and mRNA expression in the adipose tissue of normal, hyperinsulinemic, and type 2 diabetic rhesus monkeys. AB - OBJECTIVE: We have cloned the rhesus monkey leptin receptor and examined its mRNA expression levels in the adipose tissue of monkeys to investigate the regulation of gene expression of the leptin receptor. RESEARCH METHODS AND PROCEDURES: Monkey leptin receptor cDNA was cloned by reverse transcriptase-polymerase chain reaction (RT-PCR). Tissue distribution of monkey leptin receptor was examined by Northern blot analysis and RT-PCR. The mRNA levels of monkey leptin receptor in adipose tissue of normal (n=10), hyperinsulinemic obese (n=8), and type 2 diabetic monkeys (n=8) were measured by quantitative RT-PCR. RESULTS: Monkey leptin receptor cDNA had at least two alternatively spliced isoforms (long and short forms). The long form of the leptin receptor mRNA was expressed relatively highly in liver, adipose tissue, hypothalamus, and choroid plexus, whereas the total leptin receptors were expressed in every tissue examined. The mRNA levels of the long form of the leptin receptor in adipose tissue were not correlated to body weight, fasting plasma insulin, plasma glucose, or plasma leptin levels. The mRNA levels of the long form of the leptin receptor were highly correlated to that of the total leptin receptor (long and short form). DISCUSSION: The long form of leptin receptor mRNA existed in adipose tissue as well as in liver and hypothalamus, suggesting that the leptin receptor in adipose tissue may be functional in adipose tissue. The expression of the leptin receptor mRNA in adipose tissue is not affected by obesity, hyperinsulinemia, or diabetes. PMID- 9738552 TI - Increased expression of complement component C3 in the plasma of obese Zucker fa and LA/N fa(f) rats compared with their lean counterparts. AB - OBJECTIVES: The objectives of this study were to determine whether there are differences in the electrophoretic profiles of plasma proteins from lean and obese rats and to identify a protein that was found to be more abundant in the plasma of obese rats. RESEARCH METHODS AND PROCEDURES: Plasma proteins from lean and obese Zucker fa and LA/N fa(f) rats were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The identity of a band that was differentially expressed was determined by amino acid sequencing and Western blot analysis. RESULTS: A band migrating approximately the same distance as the 116 kDa molecular weight marker was more prominent in plasma from obese rats than in plasma of lean rats. Partial sequencing of the peptide revealed that 17 of the first 18 amino acids at the amino terminus were identical with the corresponding residues in the alpha-chain of complement component C3. Western blot analysis confirmed the identity of the peptide as complement component C3. Complement C3 activity was measured using a hemolytic assay to determine whether there was a corresponding increase in the biological activity of this component in the serum of obese rats. Serum from obese rats was found to have 1.8 times as much complement component C3 activity as serum from lean rats. DISCUSSION: Elevated levels of complement C3 in genetically obese rats may be relevant because increased amounts of C3 could serve as a reservoir from which increased amounts of acylation stimulating protein, a cleavage product of complement C3, could be produced. PMID- 9738553 TI - Abnormally high nourishment during sensitive periods results in body weight changes across generations. AB - OBJECTIVE: This study asked whether a brief period of overnutrition during a developmentally sensitive time could impact the individual's adult weight and that of succeeding generations. RESEARCH METHODS AND PROCEDURES: Female rat pups (F1 generation) were randomly assigned to 1 of 3 groups: (1) a control group that was naturally reared by mothers; (2) another control group implanted with chronic gastric fistulas on postnatal day 4 and fed enough formula to match the growth of the mother-reared group; and (3) an experimental group gastrostomized and infused from day 8 through day 16 with a greater quantity of food than gastrostomy-reared controls (OF). On postnatal day 16, both gastrostomy-reared groups were returned to normal litters. Adult F1 females from overfed and mother-reared groups were bred with normal males to yield an F2 generation. F2 adult females were bred to normal males to produce an F3 generation. RESULTS: When adult, the F1 experimental group was heavier than control groups. F2 adults from OF mothers were smaller than those from the control group. F3 animals from OF grandmothers were heavier at weaning than F3 descendants from mother-reared animals. DISCUSSION: Excess nourishment during a developmentally sensitive period changed the metabolic phenotype of one generation so dramatically that the gestational development and subsequent phenotype of two succeeding generations were also changed. The experiment models fetal effects of gestational diabetes in humans and may help to elucidate how, independent of genetic anomalies, secular changes can be detected across generations. PMID- 9738554 TI - Assessment of body image disturbance in obesity. AB - Disturbance in body image has long been noted as one of the most distressing psychological factors for obese individuals. Yet, only in recent years have researchers and clinicians begun to appreciate the need to integrate systematic evaluation of this component into intervention programs. Accordingly, this mini review offers an overview of selected strategies for the assessment of body image disturbance in obesity. PMID- 9738555 TI - Initial therapy for chronic myelogenous leukemia: playing the odds. AB - For patients with newly diagnosed CML, the life expectancy is much better than it was 5 years ago. However, with improvements in both transplant and nontransplant therapy comes controversy over optimal first-line therapy. We argue that an evidence-based analysis that weighs the likelihoods of surviving transplantation and responding to interferon should help guide selection of initial therapy. Such a strategy would aggressively triage appropriate patients to curative therapy while advising those who are unlikely to do well with transplantation to elect less toxic therapy, which still confers a survival benefit. We realize that many other factors beside the ones mentioned here affect a clinician's recommendation for initial therapy and a patient's ultimate choice. Many extenuating circumstances, coexisting medical conditions, and personal values need to be a part of the ultimate decision. However, we hope that this guideline summarizes the current evidence and offers a rational approach to help guide newly diagnosed patients. PMID- 9738556 TI - The evolution of cancer care for children and adults. PMID- 9738557 TI - Prophylactic isolated limb perfusion for localized, high-risk limb melanoma: results of a multicenter randomized phase III trial. European Organization for Research and Treatment of Cancer Malignant Melanoma Cooperative Group Protocol 18832, the World Health Organization Melanoma Program Trial 15, and the North American Perfusion Group Southwest Oncology Group-8593. AB - PURPOSE: Patients with primary cutaneous melanoma > or = 1.5 mm in thickness are at high risk of having regional micrometastases at the time of initial surgical treatment. A phase III international study was designed to evaluate whether prophylactic isolated limb perfusion (ILP) could prevent regional recurrence and influence survival. PATIENTS AND METHODS: A total of 832 assessable patients from 16 centers entered the study; 412 were randomized to wide excision (WE) only and 420 to WE plus ILP with melphalan and mild hyperthermia. Median age was 50 years, 68% of patients were female, 79% of melanomas were located on a lower limb, and 47% had a thickness > or = 3 mm. RESULTS: Median follow-up duration is 6.4 years. There was a trend for a longer disease-free interval (DFI) after ILP. The difference was significant for patients who did not undergo elective lymph node dissection (ELND). The impact of ILP was clearly on the occurrence-as first site of progression - of in-transit metastases (ITM), which were reduced from 6.6% to 3.3%, and of regional lymph node (RLN) metastases, with a reduction from 16.7% to 12.6%. There was no benefit from ILP in terms of time to distant metastasis or survival. Side effects were higher after ILP, but transient in most patients. There were two amputations for limb toxicity after ILP. CONCLUSION: Prophylactic ILP with melphalan cannot be recommended as an adjunct to standard surgery in high-risk primary limb melanoma. PMID- 9738558 TI - Correlation of specific immune responses with survival in melanoma patients with distant metastases receiving polyvalent melanoma cell vaccine. AB - PURPOSE: The mechanisms that underlie the clinical efficacy of melanoma vaccines are not well understood. We hypothesized that the type and strength of the immune response generated by CancerVax (John Wayne Cancer Institute, Santa Monica, CA), a polyvalent melanoma cell vaccine (PMCV), might be correlated with its effect on overall survival (OS). PATIENTS AND METHODS: Seventy-seven patients began PMCV therapy after complete surgical resection of distant metastatic melanoma. During the first two treatments, PMCV was administered with bacille Calmette-Guerin (BCG). Blood was drawn at 0, 2, 4, 8, and 12 weeks to measure serum titers of immunoglobulin G (IgG) and IgM antibodies against a tumor-associated 90-kd glycoprotein antigen (TA90) expressed on most melanoma cells, including those of PMCV. Cellular immune response to PMCV was assessed by delayed-type hypersensitivity (DTH). General immune competence was assessed by skin tests to purified protein derivative (PPD), mumps, and candida. RESULTS: Median follow-up time was 31.5 months. Within the first 12 weeks of PMCV immunotherapy, there was a significant increase in the anti-TA90 IgM (P=.0001) and IgG titers (P=.0001), and in the DTH response to PMCV (P=.0001). Univariate analysis showed that high anti-TA90 IgM titer and strong PMCV-DTH were associated with improved survival (P=.051 and .0173, respectively), whereas elevated anti-TA90 IgG was correlated with decreased survival (P=.0119). Multivariate analysis considering clinical variables and PMCV immune responses identified anti-TA90 IgM, anti-TA90 IgG, and PMCV-DTH as significant independent variables influencing survival following PMCV immunotherapy (P=.0342, .0105, and .0082, respectively). These responses to PMCV were not correlated with immune responses to BCG and therefore were not a manifestation of general immune competence for responses to unrelated antigens. The median survival time and 5-year survival rate were more than 76 months and 75%, respectively, if both anti-TA90 IgM and PMCV-DTH responses were strong (> or = 800 and > or = 7 mm, respectively; n=29); 32 months and 36%, respectively, if only one response was strong (n=35); and 19 months and 8%, respectively, if neither was strong (n=13) (P < .0001). CONCLUSION: PMCV induces both humoral and cell-mediated immune responses to melanoma-associated tumor antigens, the type and strength of which appear to be directly related to its therapeutic efficacy. PMID- 9738559 TI - Results of interleukin-2-based treatment in advanced melanoma: a case record based analysis of 631 patients. AB - PURPOSE: In patients with stage IV melanoma, durable responses have been reported with treatment regimens that involve high-dose interleukin-2 (IL-2). We analyze long-term results of 631 melanoma patients from 12 institutions who had received IL-2 alone, in combination with interferon alfa 2a or 2b (IFNalpha), or with cytotoxic drugs. METHODS: Case records that contained pretreatment parameters, response data, and updated survival information were collected. After univariate analysis, the multivariate evaluation of the impact of pretreatment parameters on response and survival was performed by logistic regression and Cox's regression, respectively. RESULTS: Patients were divided into four groups according to treatment: IL-2 alone (n=117), IL-2 and chemotherapy (n=49), IL-2 and IFNalpha (n=153), and IL-2, chemotherapy, and IFNalpha (n=312). The median survival of all patients was 10.5 months and the 2- and 5-year survival rates were 19.9% and 10.4%, respectively. Independent prognostic factors for response and survival were entirely different, treatment group being the only significant factor for response, and serum lactate dehydrogenase (LDH), metastatic site, and performance predicting survival. The addition of IFNalpha to IL-2 was associated with prolonged survival, but the effect of additional chemotherapy was less obvious. CONCLUSION: Serum LDH, metastatic site, and performance status are useful stratification factors for randomized trials in metastatic melanoma. The improved long-term survival rates observed in melanoma patients treated with IL-2/IFNalpha containing regimens are notable in contrast to the reported 5-year survival rates of 2% to 6% achieved with chemotherapy, but because selection bias cannot be ruled out, the impact of IL-2, as well as all other components of the treatment regimens, on survival needs to be confirmed in prospective randomized trials. PMID- 9738560 TI - Granulocyte-macrophage colony-stimulating factor in patients with neutropenic fever is potent after low-risk but not after high-risk neutropenic chemotherapy regimens: results of a randomized phase III trial. AB - PURPOSE: A randomized unblinded phase III trial was designed to determine the ability of granulocyte-macrophage colony-stimulating factor (GM-CSF) to accelerate recovery from febrile neutropenia induced by chemotherapy. PATIENTS AND METHODS: A total of 68 patients with febrile neutropenia following chemotherapy defined as axillary temperature greater than 38 degrees C and absolute neutrophil count (ANC) less than 1 x 10(9)/L were included. After stratification for high- and low-risk chemotherapy to induce febrile neutropenia, treatment was randomized between GM-CSF at 5 microg/kg/d or control, both being associated with antibiotics. RESULTS: GM-CSF significantly reduced the median duration of neutropenia from 6 to 3 days for ANC less than 1 x 10(9)/L(P < .001) and from 4 to 3 days for ANC less than 0.5 x 10(9)/L (P=.024), days of hospitalization required for febrile neutropenia, and duration of antibiotics during hospitalization. The greatest benefit with GM-CSF appeared for patients who had received low-risk chemotherapy, for which the median duration of ANC less than 1 x 10(9)/L was reduced from 7 to 2.5 days (P < .001) and from 4 to 2 days for ANC less than 0.5 x 10(9)/L (P=.0011), the duration of hospitalization during the study from 7 to 4 days (P=.003), and the duration on antibiotics during hospitalization from 7 to 3.5 days (P < .001). A multivariate analysis, using Cox regression, showed that variables predictive for recovery from neutropenia were GM-CSF (P=.0010) and time interval between the first day of chemotherapy and randomization (P=.030). There was no benefit for GM-CSF when high-risk chemotherapy was considered. CONCLUSION: GM-CSF significantly shortened duration of neutropenia, duration of neutropenic fever-related hospitalization, and duration on antibiotics during hospitalization when febrile neutropenia occurred after low-risk chemotherapy, but not high-risk chemotherapy. PMID- 9738561 TI - Double-blind, dose-finding study of four intravenous doses of dexamethasone in the prevention of cisplatin-induced acute emesis. Italian Group for Antiemetic Research. AB - PURPOSE: A 5-hydroxytryptamine 3 (5-HT3) receptor antagonist plus dexamethasone is the most efficacious antiemetic prophylactic treatment for the prevention of cisplatin-induced acute emesis, but the optimal intravenous (i.v.) dose of dexamethasone is unknown. This prompted us to perform a multicenter, randomized, double-blind, dose-finding study that compared four different doses of dexamethasone. PATIENTS AND METHODS: Patients were randomized to receive dexamethasone, either 4, 8, 12, or 20 mg, administered by 15-minute i.v. infusion 45 minutes before cisplatin. Ondansetron 8 mg was added to dexamethasone and was administered i.v. 30 minutes before cisplatin. From March 1996 to July 1997, 531 patients were enrolled onto the study and 530 were assessable according to the intention-to-treat principle (133 patients received 4 mg; 136 patients, 8 mg; 130 patients, 12 mg; and 131 patients, 20 mg of dexamethasone). RESULTS: Complete protection from acute vomiting and nausea was achieved by 69.2% and 60.9% of patients, respectively, who received 4 mg of dexamethasone, by 69.1% and 61.0% of those who received 8 mg, by 78.5% and 66.9% of those who received 12 mg, and by 83.2% and 71.0% of those who received 20 mg of dexamethasone. Complete protection from vomiting was significantly superior in patients who received 20 mg compared with those who received 4 and 8 mg of dexamethasone (P < .005) and was superior, but not significantly, compared with those who received 12 mg. Complete protection from nausea was superior, but not significantly, in patients who received 20 mg of dexamethasone. Multifactorial analysis confirmed these results. Antiemetic treatment was well tolerated, and no significant difference was found among the four groups in the incidence of adverse events. CONCLUSION: A 20-mg single i.v. dose of dexamethasone should be considered the most efficacious prophylactic dose for the prevention of cisplatin-induced acute emesis. PMID- 9738562 TI - Open, randomized, multicenter trial of raltitrexed versus fluorouracil plus high dose leucovorin in patients with advanced colorectal cancer. Tomudex Colorectal Cancer Study Group. AB - PURPOSE: To compare raltitrexed (Tomudex; Zeneca Pharmaceuticals Ltd, Macclesfield, United Kingdom) a direct, specific thymidylate synthase (TS) inhibitor with fluorouracil (5-FU) plus high-dose leucovorin (LV) as first-line treatment for advanced colorectal cancer (ACC). PATIENTS AND METHODS: A total of 495 patients were randomized to raltitrexed (3 mg/m2) once every 3 weeks or 5-FU (400 mg/m2) plus LV (200 mg/m2) daily for 5 days every 4 weeks. RESULTS: The randomized groups were well balanced demographically. With a minimum 17-month follow-up, median survival was comparable between groups (10.9 months raltitrexed v 12.3 months 5-FU/LV; hazards ratio, 1.15; 95% confidence interval [CI], 0.93 to 1.42; P=.197), although time to progression was statistically significantly shorter in the raltitrexed group. Overall objective responses were comparable (19% raltitrexed v 18% 5-FU/LV), with more than 50% of patients in each group having stable disease. Significantly less World Health Organization (WHO) grade 3 and 4 stomatitis (2% v 16%, P < .001) and a reduced incidence of leukopenia (6% v 13%) and diarrhea (10% v 19%) occurred in the raltitrexed group (particularly at cycle 1 ). This resulted in fewer dose reductions at cycle 2 (4% raltitrexed v 28% 5-FU/LV) and early quality-of-life (QoL) benefits for raltitrexed patients. Reversible, clinically insignificant increases in transaminases were reported in 13% of raltitrexed patients. Palliative benefits of weight gain, improved performance status, and reduced disease-related symptoms were evident in both groups. CONCLUSION: Raltitrexed is confirmed as an effective option in the first line palliative management of ACC, with comparable efficacy to and tolerability advantages (in terms of reduced incidence of stomatitis, diarrhea, and leukopenia) over 5-FU/LV. Raltitrexed has the added convenience of an every 3 weeks dosing schedule. PMID- 9738563 TI - Phase II trial of nitrogen mustard, vincristine, and procarbazine in patients with recurrent glioma: North Central Cancer Treatment Group results. AB - PURPOSE: Previous investigators have reported responses in 52% of patients treated with mechlorethamine (nitrogen mustard), vincristine, and procarbazine (MOP) for recurrent glioma. To confirm these promising results, we conducted a phase II prospective study. PATIENTS AND METHODS: Sixty-three patients with histologic confirmation of recurrent glioma were treated with the MOP regimen. Patients with or without prior chemotherapy received nitrogen mustard 3 mg/m2 or 6 mg/m2, respectively, intravenously on days 1 and 8 plus vincristine 2 mg/m2 intravenously on days 1 and 8, and procarbazine 100 mg/m2 orally on days 1 to 14. Cycles were repeated every 28 days. RESULTS: Of 61 patients assessable for response, eight responded (13%), with one complete response (CR). Responses were as follows: low-grade gliomas, 19%; anaplastic astrocytomas, 11%; anaplastic oligodendrogliomas or oligoastrocytomas, 25%; and glioblastomas, 4.3%. The most common toxicity was myelosuppression with leukocyte nadirs less than 1,000/microL in 23% and platelet nadirs less than 25,000/microL in 13% of patients. Two patients died of infection in the setting of neutropenia. Nonhematologic toxicity included neurosensory changes in 21% of patients (severe in 3%) and severe dermatologic reactions in 8%. In multivariate analysis, Eastern Cooperative Oncology group (ECOG) performance status (PS) was the best predictor for response to chemotherapy (P=.01) and time to progression (P=.008), while PS and grade were the most important predictors of survival (P=.002 and .05, respectively). CONCLUSION: This study did not confirm the high response rate previously reported in recurrent gliomas. Patients with recurrent anaplastic oligodendrogliomas or oligoastrocytomas and recurrent low-grade gliomas had the highest response rates (25% and 19%, respectively). In multivariate analysis, ECOG PS was the best predictor of response, while PS and tumor grade were the most important predictors of survival. PMID- 9738564 TI - Phase I and pharmacologic study of estramustine phosphate and short infusions of paclitaxel in women with solid tumors. AB - PURPOSE: We sought to determine the tolerance of estramustine phosphate (EMP) combined with a 3-hour paclitaxel infusion in women with solid paclitaxel pretreated solid tumors. Paclitaxel pharmacology was to be studied at the recommended phase II dose. PATIENTS AND METHODS: Paclitaxel was administered to cohorts of at least three assessable patients at doses of 150, 180, 210, and 225 mg/m2, while EMP was given at 900 and 1,200 mg/m2/d in divided doses orally for 2 days preceding and on the day of paclitaxel. The pharmacologic study was performed at 225 mg/m2 paclitaxel given in the absence and 3 weeks later in the presence of EMP 900 mg/m2/d. RESULTS: Thirty-eight patients received a total of 178 courses. Grade 3 nausea, vomiting, and diarrhea were common at EMP 1,200 mg/m2 and paclitaxel 225 mg/ m2; this was considered the maximum-tolerated dose. Since these toxicities appeared related to EMP, the pharmacologic study used a dose of 900 mg/m2 of this agent with 225 mg/m2 paclitaxel. Antitumor activity was documented against breast and ovarian cancers at all levels. Paclitaxel pharmacokinetics without and with EMP did not differ. CONCLUSION: EMP 900 mg/m2 for 3 days and 225 mg/m2 paclitaxel by 3-hour infusion are well tolerated; antitumor activity was seen in women with paclitaxel-pretreated solid tumors. This apparent enhancement of antitumor effects is unlikely to be mediated by P glycoprotein. PMID- 9738565 TI - Phase I and pharmacokinetic study of paclitaxel in combination with biricodar, a novel agent that reverses multidrug resistance conferred by overexpression of both MDR1 and MRP. AB - PURPOSE: To evaluate the feasibility of administering biricodar (VX-710; Incel, Vertex Pharmaceuticals Inc, Cambridge, MA), an agent that modulates multidrug resistance (MDR) conferred by overexpression of both the multidrug resistance gene product (MDR1) P-glycoprotein and the MDR-associated protein (MRP) in vitro, in combination with paclitaxel. The study also sought to determine the maximum tolerated dose (MTD) of paclitaxel that could be administered with biologically relevant concentrations of VX-710 and characterize the toxicologic and pharmacologic profiles of the VX-710/ paclitaxel regimen. PATIENTS AND METHODS: Patients with solid malignancies were initially treated with VX-710 as a 24-hour infusion at doses that ranged from 10 to 120 mg/m2 per hour. After a 2-day washout period, patients were re-treated with VX-710 on an identical dose schedule followed 8 hours later by paclitaxel as a 3-hour infusion at doses that ranged from 20 to 80 mg/m2. The pharmacokinetics of both VX-710 and paclitaxel were studied during treatment with VX-710 alone and VX-710 and paclitaxel. Thereafter, patients received VX-710 and paclitaxel every 3 weeks. RESULTS: VX 710 alone produced minimal toxicity. The toxicologic profile of the VX 710/paclitaxel regimen was similar to that reported with paclitaxel alone; neutropenia that was noncumulative was the principal dose-limiting toxicity (DLT). The MTD levels of VX-710/ paclitaxel were 120 mg/m2 per hour and 60 mg/m2, respectively, in heavily pretreated patients and 120/60 to 80 mg/m2 per hour in less heavily pretreated patients. At these dose levels, VX-710 steady-state plasma concentrations (Css) ranged from 2.68 to 4.89 microg/mL, which exceeded optimal VX-710 concentrations required for MDR reversal in vitro. The pharmacokinetics of VX-710 were dose independent and not influenced by paclitaxel. In contrast, VX-710 reduced paclitaxel clearance. At the two highest dose levels, which consisted of VX-710 120 mg/m2 per hour and paclitaxel 60 and 80 mg/m2, pertinent pharacokinetic determinants of paclitaxel effect were similar to those achieved with paclitaxel as a 3-hour infusion at doses of 135 and 175 mg/m2, respectively. CONCLUSION: VX-710 alone is associated with minimal toxicity. In combination with paclitaxel, biologically relevant VX-710 plasma concentrations are achieved and sustained for 24 hours, which simulates optimal pharmacologic conditions required for MDR reversal in vitro. The acceptable toxicity profile of the VX-710/ paclitaxel combination and the demonstration that optimal pharmacologic conditions for MDR reversal are achievable support a rationale for further trials of VX710/paclitaxel in patients with malignancies that are associated with de novo or acquired resistance to paclitaxel caused by overexpression of MDR1 and/or MRP. PMID- 9738566 TI - Phase I and pharmacologic study of intermittent twice-daily oral therapy with capecitabine in patients with advanced and/or metastatic cancer. AB - PURPOSE: Capecitabine is an orally administered fluoropyrimidine carbamate selectively activated to fluorouracil (5-FU) in tumors. It passes through the intestinal mucosal membrane intact and is subsequently activated by a cascade of three enzymes that results in the preferential release of 5-FU at the tumor site. PATIENTS AND METHODS: In this phase I study, capecitabine was administered twice daily as outpatient therapy, each cycle administered for 2 weeks followed by 1 week of rest. Thirty-four patients with solid tumors, all of whom except three patients were pretreated, were treated at dose levels from 502 to 3,514 mg/m2 daily. RESULTS: The median treatment duration was four cycles (85 days; range, 14 to 833+ days). Two patients continue on treatment at 686 and 833+ days. Capecitabine 3,000 mg/m2 daily was not tolerable, with dose-limiting toxicities of diarrhea with hypotension, abdominal pain, and leukopenia. Palmar-plantar erythrodysesthesia (PPE) became evident at higher dose levels after prolonged treatment. Evidence of objective tumor response was reported in four patients at 2,510 mg/m2 daily and greater (one complete response [CR] and three partial responses [PRs]) with subjective minor tumor responses in a further seven patients. Pharmacokinetic studies showed rapid gastrointestinal absorption of capecitabine, followed by extensive conversion into 5'-deoxy-5-fluorouridine (5' DFUR), with only low systemic 5-FU levels. CONCLUSION: Capecitabine is a tolerable oral outpatient therapy that shows promising clinical activity in a variety of cancers. The recommended phase II dose is 2,510 mg/m2 daily administered by this intermittent schedule. PMID- 9738567 TI - Phase I trial of continuous infusion flavopiridol, a novel cyclin-dependent kinase inhibitor, in patients with refractory neoplasms. AB - PURPOSE: We conducted a phase I trial of the cyclin-dependent kinase inhibitor, flavopiridol (National Service Center [NSC] 649890), to determine the maximum tolerated dose (MTD), toxicity profile, and pharmacology of flavopiridol given as a 72-hour infusion every 2 weeks. PATIENTS AND METHODS: Seventy-six patients with refractory malignancies with prior disease progression were treated with flavopiridol, with first-cycle pharmacokinetic sampling. RESULTS: Forty-nine patients defined our first MTD, 50 mg/m2/d x 3 with dose-limiting toxicity (DLT) of secretory diarrhea at 62.5 mg/kg/d x 3. Subsequent patients received antidiarrheal prophylaxis (ADP) to define a second MTD, 78 mg/m2/d x 3 with DLT of hypotension at 98 mg/m2/d x 3. Other toxicities included a proinflammatory syndrome with alterations in acute-phase reactants, particularly at doses >50 mg/ m2/d x 3, which in some patients prevented chronic therapy every 2 weeks. In some patients, ADP was not successful, requiring dose-deescalation. Although approximately 70% of patients displayed predictable flavopiridol pharmacology, we observed unexpected interpatient variability and postinfusion peaks in approximately 30% of cases. At the two MTDs, we achieved a mean plasma flavopiridol concentration of 271 nM (50 mg/m2/d x 3) and 344 nM (78 mg/m2/d x 3), respectively. One partial response in a patient with renal cancer and minor responses (n=3) in patients with non-Hodgkin's lymphoma, colon, and renal cancer occurred. CONCLUSION: The MTD of infusional flavopiridol is 50 mg/m2/d x 3 with dose-limiting secretory diarrhea at 62.5 mg/m2/d x 3. With ADP, 78 mg/m2/d x 3 was the MTD, with dose-limiting hypotension at 98 mg/m2/d x 3. Based on chronic tolerability, 50 mg/m2/d x 3 is the recommended phase II dose without ADP. Antitumor effect was observed in certain patients with renal, prostate, and colon cancer, and non-Hodgkin's lymphoma. Concentrations of flavopiridol (200 to 400 nM) needed for cyclin-dependent kinase inhibition in preclinical models were achieved safely. PMID- 9738568 TI - Phase II trial of chemotherapy alone for primary CNS and intraocular lymphoma. AB - PURPOSE: Primary CNS lymphoma (PCNSL) and primary intraocular lymphoma (IOL) are usually treated with radiation therapy alone or in combination with chemotherapy. The neurotoxicity of these treatments can be substantial. This study attempts to define the toxicity and efficacy of the treatment of this disease with chemotherapy alone. PATIENTS AND METHODS: Fourteen nonimmunocompromised patients were accrued to a chemotherapy regimen that incorporated a 24-hour infusion of high-dose methotrexate total dose of 8.4 g/m2 with leucovorin rescue; thiotepa 35 mg/m2; vincristine 1.4 mg/m2; dexamethasone; and intrathecal cytarabine (Ara-C) and methotrexate (MTV) administered in 21-day cycles. Seven patients were prospectively followed up with formal neuropsychologic assessments for evidence of CNS toxicity. RESULTS: The response rate was 100% with 11 (79%) complete responses and three (21%) partial responses. Cumulative survival and progression free survival rates at more than 4.5 years were 68.8% and 34.3%, respectively. Median survival has not been reached, and median progression-free survival was 16.5 months. Toxicity included severe leukoencephalopathy that was clearly attributable to chemotherapy (two patients), grade 3 or 4 neutropenia in 50% of the cycles administered, ileus (one patient), and seizures (two patients). Mucositis and renal and hepatic toxicity were mild and not therapy limiting. CONCLUSION: The MTV regimen is generally well tolerated and produces a high complete response rate. Chemotherapy alone should be investigated further in this disease to assess the necessity of initial radiation therapy, either alone or in combined modality regimens, for the achievement of optimal response and survival. PMID- 9738569 TI - Treatment of hairy cell leukemia with 2-chlorodeoxyadenosine via the Group C protocol mechanism of the National Cancer Institute: a report of 979 patients. AB - PURPOSE: To provide cladribine (CdA) to physicians for the treatment of patients with previously treated or untreated hairy cell leukemia (HCL), and to determine the response rate, response duration, survival, and toxicity with this agent. PATIENTS AND METHODS: This Group C phase II study was open to all eligible patients whose primary physician obtained written permission from the National Cancer Institute (NCI) to register patients onto this protocol. Of 979 patients registered, 861 were assessable for response and 895 for toxicity. RESULTS: The complete remission (CR) rate was 50% and the partial remission (PR) rate was 37%. At a median follow-up of 52 months, 12% of patients were reported to have progressed and 62 (7%) have died of disease. CONCLUSION: This large experience confirms the excellent response rates and remission duration of CdA in patients with HCL. Nevertheless, the response rates in this setting, which approximates general clinical practice, were lower than in other series. In general, CdA was well tolerated, but the potential increased risk for secondary malignancies requires additional follow-up evaluation. CdA can now be considered as one of the best agents for the treatment of HCL. PMID- 9738570 TI - High risk of leukemia after short-term dose-intensive chemotherapy in young patients with solid tumors. AB - PURPOSE: To help fill the gap in knowledge about the risk of leukemia from repetitive high-dose use of alkylating agents and topoisomerase-II inhibitors in young patients with solid tumors. METHODS: Poor-risk solid tumors were treated with four courses of cyclophosphamide (4,200 mg/m2)/ doxorubicin (75 mg/m2), and three courses of ifosfamide (9,000 mg/m2)/etoposide (500 mg/m2). The cumulative incidence of treatment-related myelodysplasia/ leukemia (t-AML) was calculated using the method of competing risks. The expected number of leukemic events was calculated by applying national incidence rates to person-years classified by age and sex. RESULTS: Among 86 patients (median age, 17 years) monitored for 6 to 88 months (median, 24), five cases of t-AML were detected at 10 to 37 months (median, 17). The expected number of leukemic events in this cohort was .001. Clinical and cytogenetic findings implicated prior alkylator therapy in three cases and prior treatment with topoisomerase-II inhibitors in two. At 40 months, the cumulative incidence of t-AML was 8% (SE 7%). CONCLUSION: Repetitive high dose use of alkylating agents given with topoisomerase-II inhibitors is strongly leukemogenic, even with modest cumulative doses of each drug. This finding is notable for the following reasons: (1) it undermines predictions that limited use of high-dose chemotherapy might be minimally leukemogenic, and (2) it contrasts strikingly with the previously reported low risk of t-AML following treatment of pediatric solid tumors with chemotherapy lacking the alkylator dose-intensity and prominence of etoposide that are hallmarks of current regimens. PMID- 9738571 TI - Combination chemotherapy using vinblastine and methotrexate for the treatment of progressive desmoid tumor in children. AB - PURPOSE: We report the treatment of 10 children for progressive desmoid tumor not amenable to standard surgical or radiation therapy with the use of vinblastine (VBL) and methotrexate (MTX). PATIENTS AND METHODS: Ten patients aged 6.4 to 18 years with primary (two patients) or recurrent (eight patients) desmoid tumor were treated with VBL and MTX for 2 to 35 months. Patients with recurrent tumors had been previously treated with surgical resection with (two patients) or without (five patients) radiation therapy or with radiation therapy alone (one patient). No patient had previously received cytotoxic chemotherapy. The tumor response was assessed at routine intervals by physical examination and magnetic resonance imaging (MRI). RESULTS: Five patients had clinical evidence of response to therapy with complete resolution (three patients) or partial resolution (two patients) of physical examination and radiographic abnormalities. Three patients had stable disease during 10 to 35 months of treatment. Two of these patients had progressive disease 9 and 37 months after treatment stopped; one patient had no progression 16 months after therapy. Two additional patients with stable disease had chemotherapy discontinued after 2 and 3 months. Common side effects included mild alopecia and myelosuppression and moderate nausea and vomiting. In patients with responding tumors, MRI showed decreased tumor size and, in two patients, changes consistent with fibrosis and decreased cellularity of the tumor. CONCLUSION: Combination chemotherapy with VBL and MTX appears to control desmoid tumor without significant acute or long-term morbidity in most children. This may allow for further growth and development in these patients, which may decrease the morbidity of subsequent definitive therapy. PMID- 9738572 TI - Clinical, pathologic, and molecular spectrum of tumors associated with t(11;22)(p13;q12): desmoplastic small round-cell tumor and its variants. AB - PURPOSE: Intense investigation has reshaped concepts about undifferentiated tumors occurring in young people (small round-cell tumors). Tumors associated with t(11;22)(p13;q12) and descriptively designated desmoplastic small round-cell tumor (DSRCT) are a distinctive, rare, poorly understood member of this family. We reviewed 109 cases of DSRCT to further characterize this entity better. METHODS: Clinical information and histology were reviewed. Immunohistochemistry and immunoblotting were performed using standard techniques. Chimeric EWS-WT1 RNA and DNA were detected by polymerase chain reaction (PCR) and genomic translocation breakpoints mapped in a subset of cases. RESULTS: There were 90 males and 19 females from 6 to 49 years of age (mean, 22 years). A total of 103 had tumor in the abdominal cavity, four in the thoracic region, one in the posterior cranial fossa, and one in the hand. Typical histologic and immunohistochemical features were usually evident in well-sampled tumors, but variations in cellularity, stromal components, cytology, architecture, and immunoreactivity occurred. Tumor cells were usually reactive with antibodies to keratin (67 of 78 cases, 86%), epithelial membrane antigen (50 of 54, 93%), vimentin (64 of 66, 97%), desmin (70 of 78, 90%), neuron-specific enolase (60 of 74, 81%), and the EWS-WT1 chimeric protein (25 of 27, 93%); typically nonreactive for muscle common actin (one of 58, 2%), myogenin (zero of eight, 0%), and chromogranin (one of 46, 2%); and variably reactive for MIC2 (nine of 47, 20%) and p53 (five of 17 with > 20% tumor cells reactive). Functional EWS-WT1 gene fusion was evident in 25 of 26 cases with genomic breakpoints in WT1 intron 7, and EWS introns 7, 8, and 9. Prognosis in general is poor, but tumors are responsive to aggressive therapy. CONCLUSION: This large review identifies a greater degree of clinical, pathologic, and molecular variation than originally appreciated for tumors associated with t(11;22)(p13;q12). Translocation and functional fusion of the EWS and WT1 genes appears to be a consistent feature of this unique tumor. PMID- 9738573 TI - Phase I study of temozolomide in children and adolescents with recurrent solid tumors: a report from the Children's Cancer Group. AB - PURPOSE: The Children's Cancer Group conducted a phase I trial of temozolomide stratified by prior craniospinal irradiation (CSI). PATIENTS AND METHODS: Children and adolescents with recurrent or progressive cancer were enrolled. Temozolomide was administered orally daily for 5 days, with subsequent courses administered every 21 to 28 days after full hematologic recovery. Dose levels tested included 100, 150, 180, 215, 245, and 260 mg/m2 daily. RESULTS: Twenty seven patients on the non-CSI stratum were assessable for hematologic toxicity. During the first three dose levels (100, 150, and 180 mg/m2 daily), only grades 1 and 2 hematologic toxicity occurred. One patient at 215 mg/m2 daily had grade 3 hematologic toxicity. Three of eight patients (38%) treated at 245 to 260 mg/m2 daily had dose-limiting toxicity (DLT), which included both neutropenia and thrombocytopenia. Twenty-two patients on the CSI stratum were assessable for hematologic toxicity. Hematologic DLT occurred in one of six patients (17%) at 100 mg/m2 daily and in two of four patients (50%) at 215 mg/m2 daily. No nonhematologic DLT occurred; nausea and vomiting occurred in more than half of the patients. After two courses of temozolomide, 10 patients had stable disease (SD), and three patients had a partial response (PR), one of whom subsequently had a complete response (CR) that persists through 24 months of follow-up. CONCLUSION: The maximum-tolerated dose (MTD) of temozolomide for children and adolescents without prior CSI is 215 mg/m2 daily and for those with prior CSI is 180 mg/m2 daily for 5 days, with subsequent courses that begin on day 28. Temozolomide is well tolerated and should undergo phase II testing in children and adolescents. PMID- 9738574 TI - Ewing's tumors with primary lung metastases: survival analysis of 114 (European Intergroup) Cooperative Ewing's Sarcoma Studies patients. AB - PURPOSE: To analyze event-free survival (EFS) and prognostic factors in patients who present with Ewing's tumors (ET) of bone and synchronous pulmonary and/or pleural metastases (ppm). PATIENTS AND METHODS: Of 1,270 patients (pts) registered at the continental office of the German/European Intergroup Cooperative Ewing's Sarcoma Studies (CESS81, CESS86, EICESS92), 114 were diagnosed ET with ppm. Patients underwent neoadjuvant therapy and local treatment of the primary tumor. Whole-lung irradiation 15 to 18 Gy was applied to 75 ppm pts. EFS and 95% confidence intervals (CIs) were estimated according to the Kaplan-Meier method, and prognostic factors were analyzed by log-rank tests and Cox and logistic regression procedures. RESULTS: On November 1, 1997, at a median time under study of 5.9 years, the 5-year EFS was 0.36 (95% CI, 0.26 to 0.46) and the 10-year EFS was 0.30 (95% CI, 0.19 to 0.41). Thirty-seven of 59 (63%) first relapses involved lung and/or pleura, and the lungs were the only site of relapse in 26 of 59 (44%) ppm-pts. Risk factors identified in univariate and multivariate tests were poor response of the primary tumor toward chemotherapy, metastatic lesions in both lungs, and treatment without additional lung irradiation. CONCLUSION: Chemotherapy response of the primary tumor is a prognostic factor in patients with ET with ppm. Strategies of treatment intensification warrant further evaluation. PMID- 9738575 TI - Anti-G(D2) antibody treatment of minimal residual stage 4 neuroblastoma diagnosed at more than 1 year of age. AB - PURPOSE: To eradicate minimal residual disease with anti-G(D2) monoclonal antibody 3F8 in stage 4 neuroblastoma (NB) diagnosed at more than 1 year of age. PATIENTS AND METHODS: Thirty-four patients were treated with 3F8 at the end of chemotherapy. Most had either bone marrow (n=31) or distant bony metastases (n=29). Thirteen patients were treated at second or subsequent remission (group I) and 12 patients in this group had a history of progressive/persistent disease after bone marrow transplantation (BMT); 21 patients were treated in first remission following N6 chemotherapy (group II). RESULTS: Before 3F8 treatment, 23 patients were in complete remission CR, eight in very good partial remission (VGPR), one in partial remission (PR), and two had microscopic foci in marrow. Twenty-five had evidence of NB by at least one measurement of occult/minimal tumor (iodine 131[(131)I]-3F8 imaging, marrow immunocytology, or marrow reverse transcriptase polymerase chain reaction [RT-PCR]). Acute self-limited toxicities of 3F8 treatment were severe pain, fever, urticaria, and reversible decreases in blood counts and serum complement levels. There was evidence of response by immunocytology (six of nine), by GAGE RT-PCR (seven of 12), and by (131)I-3F8 scans (six of six). Fourteen patients are alive and 13 (age 1.8 to 7.4 years at diagnosis) are progression-free (40 to 130 months from the initiation of 3F8 treatment) without further systemic therapy, none with late neurologic complications. A transient anti-mouse response or the completion of four 3F8 cycles was associated with significantly better survival. CONCLUSION: Despite high-risk nature of stage 4 NB, long-term remission without autologous (A)BMT can be achieved with 3F8 treatment. Its side effects were short-lived and manageable. The potential benefits of 3F8 in consolidating remission warrant further investigations. PMID- 9738576 TI - Detection of K-ras point mutations in sputum from patients with adenocarcinoma of the lung by point-EXACCT. AB - PURPOSE: Kirsten ras (K-ras) point mutations are found in 30% to 56% of pulmonary adenocarcinomas by means of highly sensitive techniques. Recently, the Point EXACCT (point mutation detection using exonuclease amplification coupled capture technique) method was described, which detected one cell with a mutation in 15,000 normal cells. The aim of this study was to examine whether K-ras point mutations could be found with this rapid method in the sputum of patients with adenocarcinoma of the lung. PATIENTS AND METHODS: DNA from paraffin-embedded adenocarcinoma and corresponding sputum samples were analyzed for mutations of the K-ras gene. Twenty-eight biopsy specimens and 54 sputum samples of 22 patients were used for amplification and K-ras codon 12 point mutation detection. RESULTS: In 11 of 22 patients (50%), a mutation in K-ras codon 12 was shown in the tumor sample. In five of 11 patients (45%) with a K-ras mutation in the tumor, the same type of mutation was identified in at least one sputum sample. A mutation could not be detected in any of the sputum samples from patients with a K-ras-negative tumor. Time between K-ras point mutation detection in sputum and clinical diagnosis of lung cancer varied from 1 month to almost 4 years. In two of the five patients with K-ras-positive sputum specimens, malignant cells were found with cytologic examination. CONCLUSION: Point-EXACCT is suitable for the detection of K-ras point mutations in sputum samples of patients with adenocarcinoma of the lung. This approach may be an important adjunct to cytology in the early diagnosis of lung cancer. PMID- 9738577 TI - Clinical relevance of cyclin D1 protein overexpression in laryngeal squamous cell carcinoma. AB - PURPOSE: To investigate the prognostic relevance of cyclin D1 gene overexpression in laryngeal squamous cell carcinomas (LSCCs). PATIENTS AND METHODS: The overexpression of cyclin D1 was analyzed in 149 LSCC patients with a median follow-up duration of 60 months using the DCS6 monoclonal antibody; only cases that overexpressed cyclin D1 in more than 5% of neoplastic cells were considered positive. RESULTS: Forty-eight cases (32.2%) were immunoreactive to the DCS6 antibody. Cyclin D1 overexpression was significantly associated with tobacco smoking and alcohol consumption, tumor extension, advanced clinical stage, and the presence of lymph node metastases. Univariate analysis showed that a shorter disease-free and overall survival were significantly associated with supraglottic site, tumor extension, advanced clinical stage, and cyclin D1 overexpression. At multivariate analysis, tumor extension and cyclin D1 overexpression were significantly associated with tumor recurrence, whereas tumor extension, supraglottic site and, at a borderline level of statistical significance, cyclin D1 overexpression, were associated with reduced overall survival. CONCLUSION: The overexpression of cyclin D1 in LSCC is associated with unfavorable clinicopathologic features and represents an independent significant predictor of laryngeal carcinoma prognosis, particularly for disease-free survival. This indicates that cyclin D1 evaluation may be a further useful element for selecting subgroups of patients who should be treated with more aggressive therapies. PMID- 9738578 TI - Southwest Oncology Group phase II trial of concurrent carboplatin, etoposide, and radiation for poor-risk stage III non-small-cell lung cancer. AB - PURPOSE: A phase II study was conducted by the Southwest Oncology Group (SWOG) to assess the efficacy and toxicity of concurrent carboplatin, etoposide, and thoracic radiation (XRT) in a defined population of poor-risk patients with stage III non-small-cell lung cancer (NSCLC). PATIENTS AND METHODS: Patients with stage III NSCLC were eligible if they were excluded from cisplatin-based protocols because of poor pulmonary or renal function, history of congestive heart failure, hearing loss, peripheral neuropathy, or weight loss. Carboplatin 200 mg/m2 daily intravenously days 1, 3, 29, and 31 and etoposide 50 mg/m2 daily intravenously days 1 through 4 and 29 through 32 were administered. Beginning day 1, XRT was delivered at 1.8 to 2.0 Gy daily to a total dose of 61 Gy. RESULTS: Within a period of 1 year, 63 patients were registered and 60 were eligible. Patient characteristics were age 47 to 79 years, performance status 0 to 1 (82%) and 2 (18%), and stages IIIA (60%) and IIIB (40%) NSCLC. The most common grades 3 and 4 toxicities included leukopenia (50%), thrombocytopenia (23%), and esophagitis (15%). There were no treatment-related deaths. The overall confirmed response rate was 29%, and median overall survival was 13 months (95% confidence interval, 11 to 14 months). The 2-year survival rate was 21%. CONCLUSION: This chemoradiotherapy regimen is well tolerated in poor-risk patients and yields a median survival similar to that of good-risk patients who received cisplatin based chemoradiotherapy. This chemoradiotherapy regimen will be compared with XRT alone in poor-risk patients with stage III NSCLC in a randomized phase III trial. PMID- 9738579 TI - Patient-reported short-term and long-term physical and psychologic symptoms: results of the continuous hyperfractionated accelerated [correction of acclerated] radiotherapy (CHART) randomized trial in non-small-cell lung cancer. CHART Steering Committee. AB - PURPOSE: The randomized multicenter trial of continuous hyperfractionated accelerated radiotherapy (CHART) versus conventional radiotherapy for patients with non-small-cell lung cancer (NSCLC) showed a significant survival benefit to CHART (29% v 20% at 2 years, P=.004). However, an assessment of the effect on physical and psychologic symptoms is vital to balance the costs and benefits of the two treatments. METHODS: A total of 356 patients in the United Kingdom completed the Rotterdam Symptom Checklist (RSCL) and the Hospital Anxiety and Depression Scale (HADS) at 10 time points. The principal aim of the analyses was to keep the methods simple, so as to allow the presentation and interpretation of the results to be as clear as possible. This was achieved by (1) considering individual symptoms rather than symptom subscales or domains, (2) assessing short term effects (up to 3 months) and long-term effects (at 1 and 2 years) separately, and (3) for the short-term analyses, (a) splitting the data randomly into an exploratory data set and a confirmatory data set, and (b) using two different methods of analysis: a subject-specific approach, which used the area under the curve (AUC) as a summary measure, and a group-based method, which plotted the percent of patients with moderate or severe symptoms over time. RESULTS: The results indicate that apart from CHART causing transient pain on swallowing and heartburn, there was little difference between the regimens in the short or long-term. CONCLUSION: Combining the results of the patient-assessed symptom comparisons with the clinical results indicates that CHART confers a major benefit without serious morbidity. PMID- 9738580 TI - Calculated prostate cancer volume greater than 4.0 cm3 identifies patients with localized prostate cancer who have a poor prognosis following radical prostatectomy or external-beam radiation therapy. AB - PURPOSE: Patients with palpable extraprostatic disease (T3) have a poor prostate specific antigen (PSA) failure-free (bNED) survival rate after radical prostatectomy (RP) or external-beam radiation therapy (RT). This study was performed to validate or refute the prognostic value of the previously defined calculated prostate cancer volume (cV(Ca)). PATIENTS AND METHODS: For patients with clinically localized disease (T1c,2), a Cox regression multivariable analysis was used to assess the ability of the cV(Ca) value to predict time to posttherapy PSA failure following RP or RT. RESULTS: The cV(Ca) value was a significant predictor (P < or = .0005) of time to posttherapy PSA failure in both an RP and RT data set independent of the one used to derive the cV(Ca)-based clinical staging system. In both RP- and RT-managed patients, estimates of 3-year bNED survival were not statistically different for patients with either T1c,2 disease and a cV(Ca) greater than 4.0 cm3 (RP, 27%; RT, 18%) or T3 disease (RP, 37%; RT, 34%). Despite pathologic T2 disease, the 3-year estimate of bNED survival was at most 51% in RP-managed patients with T1c,2 disease and cV(Ca) greater than 4.0 cm3. CONCLUSION: A cV(Ca) greater than 4.0 cm3 identified patients with T1c.2 disease whose bNED survival was poor after RT or RP despite pathologic T2 disease that suggests the presence of occult micrometastatic disease in many of these patients. Prospective randomized trials to evaluate the impact on survival of adjuvant systemic therapy in these high-risk patients are justified. PMID- 9738581 TI - Relation between literacy, race, and stage of presentation among low-income patients with prostate cancer. AB - PURPOSE: Diagnosis of advanced prostate cancer is a major health problem, especially among low-income men. Opportunities vary for early detection of prostate cancer for low-income black and white men because of financial, cultural, and social factors. In this study, we evaluated the association of poor literacy skills with higher rates of presentation of advanced stages of prostate cancer among low-income black and white men who received care in equal-access medical systems. PATIENTS AND METHODS: Literacy and stage at diagnosis of prostate cancer were evaluated in 212 low-income men who received medical care in Shreveport, LA, and Chicago, IL. The patients' literacy was assessed with the Rapid Estimate of Adult Literacy in Medicine (REALM), an individually administered reading screening test designed specifically for use in the medical setting. Logistic regression models were used to evaluate predictors of metastatic disease at presentation as a function of patient age, race, literacy, and city. RESULTS: Whereas black men were almost twice as likely to present with stage D prostate cancer (49.5% v 35.9%; P < .05), they were significantly more likely to have literacy levels less than sixth grade (52.3% v 8.7%; P < .001). However, after adjustment for differences in literacy, age, and city, race was not a significant predictor of advanced-stage prostate cancer. CONCLUSION: Low literacy may be an overlooked but significant barrier to the diagnosis of early stage prostate cancer among low-income white and black men. The development of culturally sensitive, low-literacy educational materials may improve patient awareness of prostate cancer and improve the frequency of diagnosis of early stage cancer. PMID- 9738582 TI - Risk-based recommendations for mammographic screening for women in their forties. AB - PURPOSE: To develop risk-based recommendations for mammographic screening for women in their 40s that take into account the woman's age, race, and specific risk factors. METHODS: We assumed that regular mammographic screening is justified for a 50-year-old woman, even one with no risk factors, and that a younger woman with an expected 1-year breast cancer incidence rate as great or greater than that of a 50-year-old woman with no risk factors would benefit sufficiently to justify regular screening. Recommendations under this criterion were based on age- and race-specific breast cancer incidence rates from the National Cancer Institute's (NCI's) Surveillance, Epidemiology, and End Results (SEER) Program; assessments of risk factors from the Breast Cancer Detection and Demonstration Project (BCDDP); and reports in the literature. RESULTS: Two methods, the exact-age procedure (EAP) and the grouped-age procedure (GAP), were developed. The less precise GAP only requires following a flow diagram. The proportion of white women recommended for screening by the EAP ranges from 10% for 40-year-old women to 95% for 49-year-old women, and the corresponding percentages for black women are 16% and 95%. The assumptions that underlie the guidelines are discussed critically. CONCLUSION: For women or physicians who prefer an individualized approach in deciding whether to initiate regular mammographic screening in the age range of 40 to 49 years, the present report offers recommendations based on individualized risk-factor data and clearly stated assumptions that have an empiric basis. These recommendations can be used to facilitate the counseling process. PMID- 9738583 TI - Low biologic aggressiveness in breast cancer in women using hormone replacement therapy. AB - PURPOSE: Hormone replacement therapy (HRT) has been associated with an increased risk for breast cancer. Cancers in women who use HRT are often less advanced, and lower mortality has been reported in those who use HRT than in nonusers. We sought to explain this by a comparison of indicators of tumor aggressiveness in patients who received HRT with those in patients who did not. PATIENTS AND METHODS: A population-based cohort of 477 postmenopausal women with breast cancer were interviewed for the use, type, and duration of HRT. Clinical variables and indicators of tumor aggressiveness (histologic grade, hormone receptors, DNA ploidy, S-phase fraction, and c-erbB-2 oncoprotein overexpression) were analyzed. RESULTS: Breast tumors from HRT users were smaller (odds ratio, 0.47; P=.005), had better histologic differentiation (P=.04), and had a lower proliferation rate (S-phase fraction, P=.009) than tumors from nonusers. These differences persisted after adjustments for age and method of diagnosis (mammography screening v self referral) by multiple logistic regression. No significant differences were observed in the estrogen (ER) or progesterone receptor content, c-erbB-2 oncogene overexpression, or axillary node involvement. A subgroup analysis showed that the tumor proliferation rates among HRT users were significantly lower only if HRT had been used at the time of diagnosis (P=.001). The type of HRT (estrogen v combination of estrogen and progesterone) was not associated with any clinical parameter or tumor phenotype. The association of HRT with lower proliferation rate and smaller tumor size was exclusively caused by ER-positive tumors (P=.0001 and P=.0035 v P > .1, respectively). CONCLUSION: The results indicate that breast cancer in women who receive HRT is biologically less aggressive than those without previous HRT. The lower cell-proliferation rate and smaller tumor size found in ER-positive tumors from current HRT users suggest a direct ER-mediated growth inhibitory effect of HRT on established breast tumors. This may at least partly explain why breast cancer in HRT users has a more favorable clinical course. PMID- 9738584 TI - Vascular endothelial growth factor is of high prognostic value in node-negative breast carcinoma. AB - PURPOSE: The prognostic value of vascular endothelial growth factor (VEGF) protein, known to stimulate endothelial growth and angiogenesis, was evaluated in node-negative breast carcinoma (NNBC) and compared with established prognostic factors. PATIENTS AND METHODS: In 525 consecutive patients with primary invasive NNBC (T1-2N0M0; tumor, node, metastasis stage), of whom 500 patients did not receive any systemic therapy, the cytosolic levels of VEGF165 were measured by using a quantitative enzyme-linked immunosorbent assay. The median follow-up was 46 months. Univariate and multivariate analyses were performed. RESULTS: VEGF level was significantly inversely correlated with estrogen receptor (ER) positivity but positively associated with tumor size and histologic grade. Patients with VEGF levels above the median value (2.40 pg/microg of DNA) showed a significantly shorter survival time (P=.0012) than patients with levels less than the median value, also when analyzed as a continuous variable (P=.0277). Tumor size, grade, and ER expression were all statistically significant for overall survival in univariate analyses (P=.0069, P=.014, and P < .001, respectively). Multivariate analysis showed that VEGF level was the strongest predictor of overall survival (P=.0199). Histologic grade was also an independent predictor of survival (P=.0477). Among the 381 patients with ER-positive tumors, a group in general considered to have a good prognosis, we found a significant reduction in survival for those with levels of VEGF greater than the median value (P=.0009). CONCLUSION: The results suggest that the level of VEGF165 protein is an independent, strong prognostic factor for survival in patients with NNBC, especially in the subgroup of patients with ER positivity. Thus, cytosolic VEGF165 might be useful to select patients for adjuvant systemic therapy. PMID- 9738586 TI - A single scale for comparing dose-intensity of all chemotherapy regimens in breast cancer: summation dose-intensity. AB - PURPOSE: To construct a single scale for comparing the dose-intensity of all chemotherapy regimens in breast cancer. MATERIALS AND METHODS: First-line single agent trials in metastatic disease were reviewed. The unit dose-intensity (UDI) that was required to produce a 30% complete response plus partial response (CR + PR) rate was determined for each drug. Randomized trials were then analyzed that prospectively tested dose-intensity. The dose-intensities of the drugs in each arm were expressed as fractions of their UDIs and added together. This yielded each arm's summation dose-intensity (SDI), which was then correlated with treatment outcomes. RESULTS: In the single-agent trials, dose-response relationships were linear when the studies covered a range of dose-intensities. In the randomized trials that tested dose-intensity in metastatic disease, response rates and median survival correlated linearly with the SDIs of the treatment arms. An increment of one SDI unit increased CR + PR rate by approximately 30%, CR rate by 10%, and median survival by 3.75 months. Metastatic disease trials were negative if the difference between the arms was less than 0.54 SDI units. Adjuvant trials that tested a dose-intensity difference of less than 0.65 SDI units were also negative. CONCLUSION: A single-agent dose-response database can be derived from historic literature that enables comparison of the dose-intensity of all combination regimens on one scale. The dose-intensity increase required to improve outcome can then be identified in earlier trials that tested that variable. SDI methodology should be tested prospectively in contemporary patients, and may be useful in guiding dosage increases beyond the conventional range. PMID- 9738585 TI - Markers of tumor angiogenesis and proteolysis independently define high- and low risk subsets of node-negative breast cancer patients. AB - PURPOSE: To compare the prognostic impact of tumor angiogenesis factors (vascular endothelial growth factor [VEGF], angiogenin, and basic fibroblast growth factor [bFGF]), tumor proteolysis factors (urokinase-type plasminogen activator [uPA] and plasminogen activator inhibitor-1 [PAI-1]), and conventional tumor markers (stage, grade, and steroid receptors) in early breast cancer. PATIENTS AND METHODS: In the primary clinical study, tumor angiogenesis and other factors were detected in frozen biopsies from 305 primary breast tumors. VEGF expression was assessed by chemiluminescence immunosorbent assay (ICMA); angiogenin, bFGF, uPA, and PAI-1 by enzyme-linked immunosorbent assay (ELISA); and steroid receptors (estrogen receptor [ER] and progesterone receptor [PgR]) by enzyme immunoassay (EIA). In the validating clinical study, another set of 190 node-negative primary breast tumor samples were collected at a separate institution. RESULTS: Univariate analysis of the primary study showed that VEGF levels were positively correlated with recurrence (P < .001). Angiogenin levels were positively correlated with disease relapse (P < .005) for the overall collective group, but not within the node-negative subset. No significant correlations were found between tumor bFGF levels and patient survival. In multivariate regression analysis, the only independent predictors of relapse-free survival (RFS) were VEGF, uPA, and lymph node status. In the validation set, the distribution of VEGF and uPA values were similar to those in the primary study; low expression of both VEGF and uPA identified patients with a < or = 20% likelihood of recurrence within 7 years. CONCLUSION: Separate primary and validating clinical studies concur that tumor VEGF level is the most important prognostic parameter among several markers of tumor angiogenesis and proteolysis. PMID- 9738587 TI - Prevalence and predictors of sexual dysfunction in long-term survivors of marrow transplantation. AB - PURPOSE: To describe the prevalence of sexual difficulties in men and women after marrow transplantation (MT), and to define medical, demographic, sexual, and psychologic predictors of sexual dysfunction 3 years after MT. PATIENTS AND METHODS: Four hundred seven adult MT patients were assessed pretransplantation. Survivors repeated measures of psychologic and sexual functioning at 1 and 3 years posttransplantation. RESULTS: Data were analyzed from 102 event-free 3-year survivors who defined themselves as sexually active. Men and women did not differ in sexual satisfaction pretransplantation. At 1 and 3 years posttransplantation, women reported significantly more sexual dysfunction than men. Eighty percent of women and 29% of men reported at least one sexual problem by 3 years after MT. No pretransplantation variables were significant predictors of 3-year sexual satisfaction for women. For men, pretransplantation variables of older age, poorer psychologic function, not being married, and lower sexual satisfaction predicted sexual dissatisfaction at 3 years (R2=.28; P < .001). Women who were more dissatisfied 3 years after MT did not receive hormone replacement therapy (HRT) at 1 -year posttransplantation and were less satisfied at 1 year, but not pretransplantation (R2=.35; P < .001). CONCLUSION: Sexual problems are significant in the lives of MT survivors, particularly for women. Although HRT before 1 year posttransplantation improves sexual function, it does not ensure sexual quality of life. Intervention for women is needed to apply hormonal, mechanical, and behavioral methods to prevent sexual difficulties as early after transplantation as possible. PMID- 9738588 TI - Tumor-suppressor p53: implications for tumor development and prognosis. AB - The p53 protein plays a central role in modulating cellular responses to cytotoxic stresses by contributing to both cell-cycle arrest and programmed cell death. Loss of p53 function during tumorigenesis can lead to inappropriate cell growth, increased cell survival, and genetic instability. p53 gene mutations occur in approximately half of all malignancies from a wide range of human tumors. In some tumor types, these p53 mutations are associated with poor prognosis and treatment failure. Based on these insights, new approaches are being developed to prevent, diagnose, and treat cancer. PMID- 9738589 TI - Recommended guidelines for the treatment of chemotherapy-induced diarrhea. AB - PURPOSE: Management of chemotherapy-induced diarrhea (CID) has customarily involved symptomatic treatment with opioids in conjunction with supportive care. Alternatively, patients refractory to conventional therapy have been given octreotide, a somatostatin analogue. Although this agent has been effective against CID, no widely accepted treatment guidelines that incorporate its use currently exist. An expert multidisciplinary panel was convened to formulate clinical practice guidelines for the treatment of CID. METHODS: The panel reviewed clinical data on the management of CID reported in the literature and analyzed currently available tools used to assess CID. Expert consensus was applied when published data were insufficient. Panel members also considered the effect of CID on quality of life and the cost-effectiveness and efficacy of different pharmacologic approaches. Effective resolution of CID and decreases in the need for supportive care or hospitalization were considered to be primary goals in the formulation of the guidelines. RESULTS: The panel formulated suggested practice guidelines for the management of CID that detail recommendations for the assessment and evaluation of diarrhea and the sequence and duration of administration of specific pharmacologic agents. CONCLUSION: The consensus of the panel was that standardized assessment and management of diarrhea is required to effectively control CID. The panel agreed that further data from a National Cancer Institute (NCI)-sponsored intergroup trial is required to determine the optimal dosage of octreotide and its cost in the treatment of cancer. The panel also agreed that further clinical research is warranted to address significant questions about the most effective way to assess and treat CID. PMID- 9738591 TI - The state of the society: a year of continued growth. PMID- 9738590 TI - Design and interpretation of clinical trials that evaluate agents that may offer protection from the toxic effects of cancer chemotherapy. AB - PURPOSE: To review the features of randomized clinical trials (RCTs) used in the development of agents that may protect against chemotherapy-induced toxicities, including trials of the cardioprotective agent dexrazoxane, hematologic growth factors, and amifostine; to suggest recommendations based on information gained from such trials and improvements in the design of ongoing and future trials. METHODS: Critical review of reports of RCTs obtained from a Medline search, references from these articles, and review of trials listed in the physician data query (PDQ) clinical trials data base. RESULTS: Several of the phase III trials did not use a format of comparing widely accepted strategies of chemotherapy with and without a protective agent. Instead, patients in the control arms of some of the trials have been exposed to more prolonged use or increased dosage of toxic chemotherapy that placed them at greater risk of the toxicity the protective agent was designed to prevent (eg, cardiotoxicity in trials of dexrazoxane, myelosuppression or thrombocytopenia in trials of growth factors). CONCLUSION: RCTs have shown clear evidence of biologic activity for the protective agents, but this does not imply therapeutic benefit as compared with alternative strategies such as avoidance of prolonged use of cardiotoxic agents or use of standard doses of chemotherapy. Ongoing and future trials of protective agents should be modified to avoid undue risk to patients. PMID- 9738592 TI - Complications of acute leukemia. Case one: congenital acute myelogenous leukemia with cutaneous involvement. PMID- 9738593 TI - Complications of acute leukemia. Case two: epidural chloroma of the lower spinal canal. PMID- 9738594 TI - Patient-physician relation and unproven therapies. PMID- 9738595 TI - Relevant clinical end points in biphosphonate trials. PMID- 9738596 TI - Late pulmonary toxicity of bleomycin. PMID- 9738597 TI - Adjuvant interferon for stage II melanoma. PMID- 9738598 TI - BRCA mutations and survival in breast cancer. PMID- 9738599 TI - Is ascitic fluid interleukin-12 an independent prognostic factor in ovarian cancer? The necessity of correction milligram P values for multiple comparisons. PMID- 9738600 TI - Fludarabine and hemolytic anemia in chronic lymphocytic leukemia. PMID- 9738601 TI - Cognition in survivors of high-grade glioma. PMID- 9738602 TI - Case of the month: making amends. Autopsy Committee of the College of American Pathologists. PMID- 9738603 TI - Chronic renal insufficiency: a diagnostic and therapeutic approach. AB - Chronic renal insufficiency ultimately culminating in end-stage renal disease requiring dialysis or transplantation is a major health problem in the United States. The first task confronting the physician caring for a patient with renal disease is to decide whether the renal insufficiency is acute or chronic. The initial differential diagnostic approach to chronic renal insufficiency consists of determining whether the patient has glomerular disease or interstitial or vascular disease on the basis of a careful history taking, urinalysis, and measurement of 24-hour protein excretion. Further refinement of diagnostic considerations often requires serologic studies, renal biopsy, or imaging the urinary tract with ultrasonography or computed tomography. Management considerations begin with the identification and correction of any acute reversible causes of renal insufficiency in patients with chronic renal disease. Recent studies have shown that effective antihypertensive therapy, especially with angiotensin-converting enzyme inhibitors, restriction of dietary protein, and excellent glycemic control in patients with diabetes, can retard the progression of chronic renal disease. Once these therapeutic strategies are in place, it is important to anticipate and treat the multiple manifestations of chronic progressive renal insufficiency: fluid overload, hyperkalemia, metabolic acidosis, abnormalities of calcium, phosphorus, and vitamin D metabolism, and anemia. PMID- 9738604 TI - Gaucher disease: recommendations on diagnosis, evaluation, and monitoring. AB - BACKGROUND: Timely diagnosis and continued monitoring of patients with type I Gaucher disease is critical because skeletal involvement can permanently disable patients and visceral organ involvement can lead to abdominal pain and secondary hematologic and biochemical complications. OBJECTIVE: To seek clinical consensus for minimum recommendations for effective diagnosis and monitoring of patients with type I Gaucher disease. PARTICIPANTS, EVIDENCE, AND CONSENSUS PROCESS: Contributing authors collaborated in quarterly meetings over a 2-year period to synthesize recommendations from peer-reviewed publications and their own medical experiences. These physicians care for most patients with Gaucher disease in the United States and serve as the US Regional Coordinators for the International Collaborative Gaucher Group Registry, the world's largest database for this disorder. CONCLUSIONS: The definitive method of diagnosis is enzyme assay of beta glucocerebrosidase activity. Schedules differ for monitoring complications of type I Gaucher disease, depending on symptoms and whether enzyme replacement therapy is used. Hematologic and biochemical involvement should be assessed by complete blood cell count, including platelets, acid phosphatase, and liver enzymes, at baseline and every 12 months in untreated patients and every 3 months and at enzyme replacement therapy changes in treated patients. Visceral involvement should be assessed at diagnosis using magnetic resonance imaging or computed tomographic scans. Skeletal involvement should be assessed at diagnosis using T1- and T2-weighted magnetic resonance imaging of the entire femora and plain radiography of the femora, spine, and symptomatic sites. Follow-up skeletal and visceral assessments are recommended every 12 to 24 months in untreated patients, and every 12 months and at enzyme replacement therapy changes in treated patients. PMID- 9738605 TI - Time trends in the use of cholesterol-lowering agents in older adults: the Cardiovascular Health Study. AB - OBJECTIVES: To describe recent temporal patterns of cholesterol-lowering medication use and the characteristics that may have influenced the initiation of cholesterol-lowering therapy among those aged 65 years or older. SUBJECTS AND METHODS: A cohort of 5201 adults 65 years or older were examined annually between June 1989 and May 1996. We added 687 African American adults to the cohort in 1992-1993. We measured blood lipid levels at baseline and for the original cohort in the third year of follow-up. We assessed the use of cholesterol-lowering drugs at each visit. RESULTS: The prevalence of cholesterol-lowering drug use in 1989 1990 was 4.5% among the men and 5.9% among the women; these figures increased over the next 6 years to 8.1% and 10.0%, respectively, in 1995-1996. There was a 4-fold increase in the use of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors during the 6 years of follow-up, from 1.9% of all participants in 1989-1990 to 7.5% in 1995-1996. The use of bile acid sequestrants, nicotinic acid, and probucol declined from initial levels of less than 1% each. Among the participants who were untreated in 1989-1990, but eligible for cholesterol-lowering therapy after a trial of dietary therapy according to the 1993 guidelines of the National Cholesterol Education Panel, less than 20% initiated drug therapy in the 6 years of follow-up, even among subjects with a history of coronary heart disease. Among participants untreated at baseline but eligible for either cholesterol-lowering therapy or dietary therapy, initiation of cholesterol-lowering drug therapy was directly associated with total cholesterol levels, hypertension, and a history of coronary heart disease, and was inversely related to age, high-density lipoprotein cholesterol levels, and difficulties with activities of daily living. Other characteristics that form the basis of the 1993 National Cholesterol Education Panel guidelines diabetes, smoking, family history of premature coronary heart disease, and total number of risk factors-were not associated with the initiation of cholesterol lowering drug therapy. CONCLUSIONS: Given the clinical trial evidence for benefit, those aged 65 to 75 years and with prior coronary heart disease appeared undertreated with cholesterol-lowering drug therapy. PMID- 9738606 TI - Benefits of influenza vaccination for low-, intermediate-, and high-risk senior citizens. AB - BACKGROUND: Vaccination rates for healthy senior citizens are lower than those for senior citizens with underlying medical conditions such as chronic heart or lung disease. Uncertainty about the benefits of influenza vaccination for healthy senior citizens may contribute to lower rates of utilization in this group. OBJECTIVE: To clarify the benefits of influenza vaccination among low-risk senior citizens while concurrently assessing the benefits for intermediate- and high risk senior citizens. METHODS: All elderly members of a large health maintenance organization were included in each of 6 consecutive study cohorts. Subjects were grouped according to risk status: high risk (having heart or lung disease), intermediate risk (having diabetes, renal disease, stroke and/or dementia, or rheumatologic disease), and low risk. Outcomes were compared between vaccinated and unvaccinated subjects after controlling for baseline demographic and health characteristics. RESULTS: There were more than 20000 subjects in each of the 6 cohorts who provided 147551 person-periods of observation. The pooled vaccination rate was 60%. There were 101 619 person-periods of observation for low-risk subjects, 15 482 for intermediate-risk, and 30 450 for high-risk subjects. Vaccination over the 6 seasons was associated with an overall reduction of 39% for pneumonia hospitalizations (P<.001), a 32% decrease in hospitalizations for all respiratory conditions (P<.001), and a 27% decrease in hospitalizations for congestive heart failure (P<.001). Immunization was also associated with a 50% reduction in all-cause mortality (P<.001). Within the risk subgroups, vaccine effectiveness was 29%, 32%, and 49% for high-, intermediate-, and low-risk senior citizens for reducing hospitalizations for pneumonia and influenza (for high and low risk, P< or =.002; for intermediate risk, P = .11). Effectiveness was 19%, 39%, and 33% (for each, P< or =.008), respectively, for reducing hospitalizations for all respiratory conditions and 49%, 64%, and 55% for reducing deaths from all causes (for each, P<.001). Vaccination was also associated with direct medical care cost savings of $73 per individual vaccinated for all subjects combined (P = .002). Estimates of cost savings within each risk group suggest that vaccination would be cost saving for each subgroup (range of cost savings of $171 per individual vaccinated for high risk to $7 for low risk), although within the subgroups these findings did not reach statistical significance (for each, P> or =.05). CONCLUSIONS: This study confirms that healthy senior citizens as well as senior citizens with underlying medical conditions are at risk for the serious complications of influenza and benefit from vaccination. All individuals 65 years or older should be immunized with this vaccine. PMID- 9738607 TI - Consultation between cardiologists and generalists in the management of acute myocardial infarction: implications for quality of care. AB - BACKGROUND: The rapid expansion of managed care in the United States has increased debate regarding the appropriate mix of generalist and specialist involvement in medical care. OBJECTIVE: To compare the quality of medical care when generalists and cardiologists work separately or together in the management of patients with acute myocardial infarction (AMI). METHODS: We reviewed the charts of 1716 patients with AMI treated at 22 Minnesota hospitals between 1992 and 1993. Patients eligible for thrombolytic aspirin, beta-blockers, and lidocaine therapy were identified using criteria from the 1991 American College of Cardiology guidelines for the management of AMI. We compared the use of these drugs among eligible patients whose attending physician was a generalist with no cardiologist input, a generalist with a cardiologist consultation, and a cardiologist alone. RESULTS: Patients cared for by a cardiologist alone were younger, presented earlier to the hospital, were more likely to be male, had less severe comorbidity, and were more likely to have an ST elevation of 1 mm or more than generalists' patients. Controlling for these differences, there was no variation in the use of effective agents between patients cared for by a cardiologist attending physician and a generalist with a consultation by a cardiologist. However, there was a consistent trend toward increased use of aspirin, thrombolytics, and beta-blockers in these patients compared with those with a generalist attending physician only (P<.05 for beta-blockers only). Differences between groups in the use of lidocaine were not statistically significant. The adjusted probabilities of use of thrombolytics for consultative care and cardiologist attending physicians were 0.73 for both. Corresponding probabilities were 0.86 and 0.85 for aspirin and 0.59 and 0.57 for beta-blockers, respectively. CONCLUSIONS: For patients with AMI, consultation between generalists and specialists may improve the quality of care. Recent policy debates that have focused solely on access to specialists have ignored the important issue of coordination of care between generalist and specialist physicians. In hospitals where cardiology services are available, generalists may be caring for patients with AMI who are older and more frail. Future research and policy analyses should examine whether this pattern of selective referral is true for other medical conditions. PMID- 9738608 TI - The AUDIT alcohol consumption questions (AUDIT-C): an effective brief screening test for problem drinking. Ambulatory Care Quality Improvement Project (ACQUIP). Alcohol Use Disorders Identification Test. AB - OBJECTIVE: To evaluate the 3 alcohol consumption questions from the Alcohol Use Disorders Identification Test (AUDIT-C) as a brief screening test for heavy drinking and/or active alcohol abuse or dependence. METHODS: Patients from 3 Veterans Affairs general medical clinics were mailed questionnaires. A random, weighted sample of Health History Questionnaire respondents, who had 5 or more drinks over the past year, were eligible for telephone interviews (N = 447). Heavy drinkers were oversampled 2:1. Patients were excluded if they could not be contacted by telephone, were too ill for interviews, or were female (n = 54). Areas under receiver operating characteristic curves (AUROCs) were used to compare mailed alcohol screening questionnaires (AUDIT-C and full AUDIT) with 3 comparison standards based on telephone interviews: (1) past year heavy drinking (>14 drinks/week or > or =5 drinks/ occasion); (2) active alcohol abuse or dependence according to the Diagnostic and Statistical Manual of Mental Disorders, Revised Third Edition, criteria; and (3) either. RESULTS: Of 393 eligible patients, 243 (62%) completed AUDIT-C and interviews. For detecting heavy drinking, AUDIT-C had a higher AUROC than the full AUDIT (0.891 vs 0.881; P = .03). Although the full AUDIT performed better than AUDIT-C for detecting active alcohol abuse or dependence (0.811 vs 0.786; P<.001), the 2 questionnaires performed similarly for detecting heavy drinking and/or active abuse or dependence (0.880 vs 0.881). CONCLUSIONS: Three questions about alcohol consumption (AUDIT-C) appear to be a practical, valid primary care screening test for heavy drinking and/or active alcohol abuse or dependence. PMID- 9738609 TI - Diabetic ketoacidosis associated with cocaine use. AB - BACKGROUND: Multiple risk factors for diabetic ketoacidosis (DKA) have been described, including omission of insulin therapy and clinical conditions known to increase counterregulatory hormones. Recently, substance abuse has been identified in patients with DKA. We observed many cases of DKA in cocaine users, although the association between cocaine use and DKA has not been well described in the medical literature. METHODS: We performed a retrospective case-control study of admissions for DKA in cocaine users and non-user controls in an urban teaching hospital from January 1, 1985, to December 31, 1994. RESULTS: We identified 720 adult admissions for DKA. Twenty-seven cocaine users accounted for 102 admissions (14% of all DKA admissions). The users were compared with 85 nonuser controls who had 154 DKA admissions. Cocaine users had more admissions for DKA (mean, 3.78 vs 1.81; P = .03). Cocaine users were less likely than controls to have an intercurrent illness identified as a precipitating factor for DKA (14.7% vs 33.1%; P<.001) and were more likely to have missed taking insulin prior to admission (45.1% vs 24.7%; P<.001). Although cocaine users had higher serum glucose levels on admission (32.9 mmol/L [593.4 mg/dL] vs 29.5 mmol/L [531.1 mg/dL]; P =.03), no differences in intensity of illness or treatment outcome were detected. CONCLUSIONS: In this preliminary study, cocaine use was found in a significant number of adults admitted with DKA and was associated with more frequent omission of insulin therapy and the absence of precipitating systemic illness. Either because of its association with insulin therapy omission or its effects on counterregulatory hormones, cocaine use should be considered a risk factor for DKA, particularly in patients with multiple admissions. PMID- 9738610 TI - Fluctuations of lipid and lipoprotein levels in hyperlipidemic postmenopausal women receiving hormone replacement therapy. AB - BACKGROUND: Fluctuations in lipid and lipoprotein levels are encountered quite often in hyperlipidemic patients. We examined the possibility that lipid and lipoprotein levels fluctuate due to the different effects of estrogen and progestogen in postmenopausal hyperlipidemic women receiving combined hormonal replacement therapy. METHODS: In an open-label study conducted during 3 consecutive hormonal cycles (3 months), levels of fasting total cholesterol, triglycerides, and low (LDLC)- and high-density lipoprotein cholesterol (HDLC) were determined in 36 postmenopausal hyperlipidemic women on day 13 of conjugated equine estrogen (1.25 mg/d) therapy and on day 25 after 12 days of receiving estrogen plus medroxyprogesterone acetate (5 mg/d). RESULTS: While receiving estrogen and combined therapies, means +/- SD total cholesterol levels increased from 6.50 +/- 0.97 mmol/L (251 +/- 37 mg/dL) to 6.88 +/- 1.42 mmol/L (266 +/- 54 mg/dL) (P<.001); LDLC levels, from 4.05 +/- 1.14 mmol/L (156 +/- 44 mg/dL) to 4.62 +/- 1.36 mmol/L (178 +/- 52 mg/dL) (P<.001). Mean +/- SD HDLC cholesterol levels decreased from 1.44 +/- 0.32 mmol/L (55 +/- 12 mg/dL) to 1.29 +/- 0.28 mmol/L (50 +/- 10 mg/dL) (P<.001); triglyceride levels, from 2.23 +/- 1.03 mmol/L (197 +/- 91 mg/dL) to 2.06 +/- 1.04 mmol/L (182 +/- 92 mg/dL) (P<.001). CONCLUSIONS: Hyperlipidemic postmenopausal women receiving combined sequential estrogen and progestogen replacement therapy demonstrate very significant fluctuations in their lipid and lipoprotein levels. These fluctuations depend on the hormonal phase, ie, estrogen alone or combined with progestogen. PMID- 9738611 TI - Expanding eligibility for outpatient treatment of deep venous thrombosis and pulmonary embolism with low-molecular-weight heparin: a comparison of patient self-injection with homecare injection. AB - BACKGROUND: The outpatient treatment of patients with deep vein thrombosis and pulmonary embolism using low-molecular-weight heparin has the potential to reduce health care costs, but it is unclear if most patients with deep vein thrombosis and pulmonary embolism can be treated as outpatients. In the published studies, more than 50% of patients were excluded from outpatient treatment for reasons such as comorbid conditions, short life expectancy, concomitant pulmonary embolism, and previous deep vein thrombosis, and many patients were not treated entirely at home. We sought to determine if expanding patient eligibility for the outpatient treatment of deep vein thrombosis and pulmonary embolism affects the safety and effectiveness of the treatment, and to determine if patient self injection compared with injections administered by a homecare nurse affected these outcomes. PATIENTS AND METHODS: We treated as outpatients all patients with deep vein thrombosis and pulmonary embolism, except for those with massive pulmonary embolism, high risk for major bleeding or an active bleed, phlegmasia, and patients hospitalized for reasons that prevented discharge. We compared 2 models of outpatient care to determine feasibility, safety, and efficacy. Both models involved nurse managers who provided daily patient contact and ongoing treatment; however, in one model the patients were taught to inject themselves and in the other model homecare nurses administered the injections. We expanded the population of patients eligible for outpatient treatment by including many patients not treated at home in previous studies. Most patients in our study were treated with dalteparin sodium, 200 U/kg every 24 hours, for a minimum of 5 days. Therapy with warfarin sodium was started on the day of diagnosis or the following day. Patients were followed up for 3 months to determine rates of recurrent venous thromboembolism, bleeding, and death. RESULTS: In this study, 194 (83%) of 233 consecutive patients were deemed eligible and treated as outpatients. Of the 39 patients who did not receive home therapy, 20 had concomitant medical problems responsible for their admission or were already inpatients, 6 had massive pulmonary embolism, 6 refused to pay for the dalteparin therapy, 4 had active bleeding, and 3 had phlegmasia cerulea dolens, which required treatment with intravenous narcotics for pain control. More than 184 (95%) of the 194 patients were treated entirely at home. There was no significant difference (P>.99) in the rate of recurrent venous thromboembolic events between the patients who were injected by homecare nurses (3/95 [3.2%]) and those who injected themselves (4/99 [4.0%]). Combining the 2 models, the overall recurrent event rate was 3.6% (95% confidence interval, 1.5%-7.4%). Similarly, there were no significant differences in rates of major hemorrhage (2/95 vs 2/99; P>.99), minor hemorrhage (8/95 vs 2/99; P = .06), and death (6/95 vs 8/99; P = .63). The overall rate of major hemorrhage was 2.0% (95% confidence interval, 0.6%-5.2%). CONCLUSIONS: We demonstrate that more than 80% of patients at our tertiary care hospital could be treated at home using 1 of the 2 models of care we describe. Our results demonstrate that patients can safely and effectively perform home self-injection under the supervision of a hospital-based nurse. Injections at home by a homecare nurse are similarly effective. Our overall rates of recurrent venous thromboembolism, bleeding, and death are at least as favorable as those previously reported despite using 1 dose per day of dalteparin for most patients. PMID- 9738613 TI - Acute exposure to aliphatic hydrocarbons: an unusual cause of acute tubular necrosis. AB - Acute tubular necrosis is not an uncommon phenomenon, but it rarely results from environmental factors. We describe a patient in whom acute renal failure developed 2 times after overexposure to aliphatic hydrocarbons and discuss some potential pathophysiological mechanisms. This association has rarely been reported in the literature. Considering the wide availability of aliphatic hydrocarbons in diesel fuel and solvents, associated renal toxicity is probably underrecognized. We stress the importance of identifying environmental and professional factors as causes of acute tubular necrosis. PMID- 9738612 TI - National trends in the use of antibiotics by primary care physicians for adult patients with cough. AB - BACKGROUND: Increased antibiotic use for outpatient illnesses has been identified as an important determinant of the recent rise in antibiotic resistance among common respiratory pathogens. Efforts to reduce the inappropriate use will need to be evaluated against current trends in the outpatient use of antibiotics. OBJECTIVES: To examine national trends in the use of antibiotics by primary care physicians in the care of adult patients with cough and identify patient factors that may influence antibiotic use for these patients. METHODS: This study was based on a serial analysis of results from all National Ambulatory Medical Care Surveys beginning in 1980 (when therapeutic drug use was first recorded) to 1994 (the most recent survey year available). These surveys are a random sampling of visits to US office-based physicians in 1980, 1981, 1985, and annually from 1989 1994. Eligible visits included those by adults presenting to general internists, family practitioners, or general practitioners with a chief complaint of cough. A total of 3416 visits for cough were identified over the survey years. Survey results were extrapolated, based on sampling weights in each year, to project national rates of antibiotic use for patients with cough. Additional analyses examined the rates of antibiotic use stratified by patient age, race, and clinical diagnosis. RESULTS: Overall, an antibiotic was prescribed 66% of the time during office visits for patients with cough: 59% of patient visits in 1980 rising to 70% of visits in 1994 (P = .002 for trend). In every study year, white, non-Hispanic patients and patients younger than 65 years were more likely to receive antibiotics compared with nonwhite patients and patients 65 years or older, respectively. CONCLUSIONS: The rate of antibiotic use by primary care physicians for patients with cough remained high from 1980 to 1994, and was influenced by nonclinical characteristics of patients. PMID- 9738614 TI - Cocaine use and chest pain syndromes. PMID- 9738615 TI - Mycobacterium avium complex lymph node abscess after use of highly active antiretroviral therapy in a patient with AIDS. PMID- 9738616 TI - Entrenched medical academia as a culprit. PMID- 9738617 TI - Voluntary, nonremunerated blood donation: still a world health goal? PMID- 9738618 TI - Banking and transplantation of unrelated donor umbilical cord blood: status of the National Heart, Lung, and Blood Institute-sponsored trial. PMID- 9738619 TI - The distribution of infectivity in blood components and plasma derivatives in experimental models of transmissible spongiform encephalopathy. AB - BACKGROUND: The administration of blood components from donors who subsequently develop Creutzfeldt-Jakob disease has raised the issue of blood as a possible vehicle for iatrogenic disease. STUDY DESIGN AND METHODS: We examined infectivity in blood components and Cohn plasma fractions in normal human blood that had been "spiked" with trypsinized cells from a scrapie-infected hamster brain, and in blood of clinically ill mice that had been inoculated with a mouse-adapted strain of human transmissible spongiform encephalopathy. Infectivity was assayed by intracerebral inoculation of the blood specimens into healthy animals. RESULTS: Most of the infectivity in spiked human blood was associated with cellular blood components; the smaller amount present in plasma, when fractionated, was found mainly in cryoprecipitate (the source of factor VIII) and fraction I+II+III (the source of fibrinogen and immunoglobulin); almost none was recovered in fraction IV (the source of vitamin-K-dependent proteins) and fraction V (the source of albumin). Mice infected with the human strain of spongiform encephalopathy had very low levels of endogenous infectivity in buffy coat, plasma, cryoprecipitate, and fraction I+II+III, and no detectable infectivity in fractions IV or V. CONCLUSION: Convergent results from exogenous spiking and endogenous infectivity experiments, in which decreasing levels of infectivity occurred in cellular blood components, plasma, and plasma fractions, suggest a potential but minimal risk of acquiring Creutzfeldt-Jakob disease from the administration of human plasma protein concentrates. PMID- 9738620 TI - Surveillance for Creutzfeldt-Jakob disease among persons with hemophilia. AB - BACKGROUND: Although Creutzfeldt-Jakob disease (CJD) has been shown to be transmissible through blood components in rodent models, no human blood-to-blood transmission has been documented. If blood transmission were possible in humans, persons with hemophilia in the United States would be at higher risk of contracting CJD, because they receive large numbers of blood components. Nearly one-half of the hemophilia population contracted HIV in the 1980s, and many of these people have since died with neurologic complications. This study investigated whether some hemophilia patients with neurologic disorders may have died with CJD. STUDY DESIGN AND METHODS: Hemophilia treatment Centers across the United States were invited to participate in this retrospective surveillance study. The centers were asked to send any available formalin-fixed paraffin block brain samples from hemophilia decedents. Slides were prepared at the Centers for Disease Control and Prevention and reviewed by three expert neuropathologists. Two slides were stained for the prion protein at the request of one of the neuropathologists. RESULTS: Specimens from 24 decedents with genetic bleeding disorders were collected and reviewed.The panel found no evidence of CJD in any of the specimens. CONCLUSIONS: Although the study sample is small, these results support the growing evidence that CJD is not being transmitted in the nation's blood supply. PMID- 9738622 TI - A prospective randomized trial of the surgical blood order equation for ordering red cells for total hip arthroplasty patients. AB - BACKGROUND: The majority of crossmatched blood is for surgical patients, and most of it is never transfused. An alternative system for ordering red cell (RBC) units, called the surgical blood order equation (SBOE), which incorporates specific patient variables for surgical patients, has been developed. STUDY DESIGN AND METHODS: A prospective double-blind randomized trial compared the SBOE with the maximal surgical blood order schedule (MSBOS) system for ordering allogeneic RBC units in 60 patients undergoing total hip arthroplasty. Autologous RBCs were available for none of the patients. RESULTS: There were no differences in patient demographic, surgical, or laboratory variables at any time. The median number (range) of allogeneic RBC units ordered was 2 (2-3) for the MSBOS and 0 (0 3) for the SBOE (p<0.0001). The SBOE ordered the correct number of RBC units for 58 percent of patients, while the MSBOS did so for 7 percent (p<0.0001). The SBOE had a lower crossmatch-to-transfusion ratio than the MSBOS (0.83 vs. 4.12). Costs were also lower with the SBOE. CONCLUSION: Incorporation of patient factors in the use of the SBOE system resulted in increased efficiency of blood-ordering practices for total hip arthroplasty. PMID- 9738621 TI - Exposure to GB virus type C or hepatitis G virus in selected Australian adult and children populations. AB - BACKGROUND: The epidemiology and disease association for the GB virus type C (GBV C) or hepatitis G virus (HGV) are poorly understood. STUDY DESIGN AND METHODS: This study describes the exposure rates to GBV-C/HGV in diverse Australian population groups by testing for current infection and evidence of past infection with a reverse transcriptase polymerase chain reaction and an anti-E2 enzyme linked immunosorbent assay, respectively. Subjects included volunteer blood donors, hepatitis C antibody (anti-HCV)-positive donors, children, hemodialysis patients, pregnant women attending a prenatal clinic, injecting drug users (IVDUs), and adult hemophiliacs. RESULTS: Combined GBV-C RNA and E2 antibody prevalence was 6.5 percent (6/93) in children, 13.3 percent (75/565) in blood donors, 14 percent (14/99) in pregnant women, 22.5 percent (18/80) in hemodialysis patients, 80 percent (56/70) in anti-HCV-positive donors, 88.6 percent (31/35) in IVDUs, and 85.7 percent (54/63) in adult hemophiliacs. Children had the lowest antibody rate, 1.1 percent, whereas the rate was 10.8 percent for blood donors and rose to 45.7 percent for IVDUs, 57.1 percent for anti-HCV-positive donors, and 74.6 percent for hemophiliacs. In contrast, current infection rates were comparable for children, blood donors, and pregnant women (5.4, 2.6, and 6%, respectively), rising to 11.1 percent for hemophiliacs, 24.3 percent for anti-HCV-positive donors, and 48.6 percent for IVDUs. Ten of 12 blood donors had persistent viremia, while 2 had recent infections, 1 with apparent resolution. CONCLUSION: Exposure to GBV-C can commence at an early age, although ongoing exposure may also occur among adults with no apparent risk factors. GBV-C RNA positivity was not associated with abnormal plasma alanine aminotransferase levels among blood donors. PMID- 9738623 TI - The use of plasma as a replacement fluid in plasma exchange. Canadian Apheresis Group. AB - BACKGROUND: The Canadian Apheresis Group has maintained a national registry of apheresis activities for the past 16 years. Since 1991, the use of plasma as a replacement fluid in plasma exchange has been recorded. STUDY DESIGN AND METHODS: Six years of data from the registry on the use of plasma as a replacement fluid were analyzed. RESULTS: Plasma was used in more than 25 percent of all plasma exchange procedures. Of 41,519 plasma exchange procedures reported, 11,970 used plasma alone or in combination with albumin. In 1991, 1026 (78%) of these procedures used plasma appropriately for either thrombotic thrombocytopenic purpura (TTP) or adult hemolytic uremic syndrome (HUS); between 1992 and 1996, these numbers were 1043 (81%), 1570 (86%), 1171 (87%), 2192 (92%), and 2741 (90%), respectively. In the remaining procedures, frozen or cryosupernatant plasma was administered to 326 patients for a total of 40 diseases other than TTP or HUS. CONCLUSIONS: In those diseases for which plasma was administered as the sole replacement fluid, no disease appears to justify such treatment without the existence of an associated condition requiring specific replenishment of some plasma component. Further evaluation of the specific indications for the use of plasma as a replacement fluid in plasma exchange is required for diseases other than TTP or HUS. PMID- 9738624 TI - Clinical factors influencing posttransfusion platelet increment in patients undergoing hematopoietic progenitor cell transplantation--a prospective analysis. AB - BACKGROUND: Platelet transfusion refractoriness remains problematic in the management of patients who have undergone hematopoietic progenitor cell transplantation. Bone marrow transplantation itself is reported to be a relevant factor hampering efficient platelet transfusions. However, a prospective analysis assessing factors affecting platelet transfusion efficacy in the setting of hematopoietic progenitor cell transplantation has yet to be conducted. STUDY DESIGN AND METHODS: To identify factors independently influencing platelet transfusion efficacy after hematopoietic progenitor cell transplantation, a prospective study was performed to determine the effectiveness of platelet transfusions by estimating posttransfusion (16-hour) corrected count increments (CCI) in 42 consecutive patients (26 who received allogeneic transplants and 16 who received autologous transplants) with 439 available platelet transfusions. RESULTS: The mean CCI and percentage of CCI <4500 for all transfusions were 6161.1 +/- 7775.2 per microL and 42.1 percent, respectively. Multiple linear regression analyses revealed high total bilirubin, total body irradiation, high serum tacrolimus, and high serum cyclosporin A to be major factors independently predicting a lower CCI. HLA antibodies with restricted specificity and platelet antibodies were detected transiently in 17 and 14 percent of the patients, respectively. The presence of these antibodies was not, however, associated with a poor response to platelet transfusions. CONCLUSION: Platelet transfusion efficacy in hematopoietic progenitor cell transplant recipients is markedly influenced by clinical factors specific to the procedure as well as those already recognized in other settings. Alloimmunization is not, however, a major factor associated with a poor response to platelet transfusions after this procedure. PMID- 9738625 TI - Evaluation of the Amicus Separator in the collection of apheresis platelets. AB - BACKGROUND: A new apheresis instrument, the Amicus Separator with software versions 2.13 and 2.34, was evaluated for component yields, collection efficiency, and incidence of donor and transfusion recipient reactions. The Amicus was also compared to the Spectra Leukocyte Reduction System (LRS) with version 5 software. STUDY DESIGN AND METHODS: Single and double apheresis platelets (APs) were collected at two locations. The targeted platelet yields were 4.0 x 10(11) for single APs and 6.8 x 10(11) for double APs. One location used a double-needle procedure, and the other used a single-needle procedure. Along with 28 of the Amicus procedures (14 at each of two locations), the same donors underwent single or double AP collections on the Spectra LRS. APs were tested for platelet yields and residual white cells. APs were transfused in three hospitals. Donor and transfusion recipient reactions and technical problems were documented. RESULTS: The Amicus Separator efficiently collected single APs (n = 59) and double APs (n = 62) with mean platelet yields of 4.2 x 10(11) and 6.5 x 10(11), respectively. When inlet line alarms occurred in single-needle procedures, platelet yields were lower and collection times were longer. All APs were white cell-reduced below 5.0 x 10(6), and all but one AP were white cell reduced below 1.0 x 10(6) without filtration. Component yields from the paired Amicus and Spectra LRS procedures were comparable. Collection times (excluding reinfusion/rinseback) were 20 to 23 minutes faster on the Amicus Separator. No serious donor or transfusion recipient reactions occurred. CONCLUSION: The Amicus Separator provided satisfactory platelet yields and collection efficiency, with shorter collection times than did the Spectra LRS, and it white cell-reduced components without filtration. PMID- 9738626 TI - Lowering the hemoglobin cutoff for female plateletpheresis donors. AB - BACKGROUND: The standards of the American Association of Blood Banks describe a minimum hemoglobin level of 12.5 g per dL for apheresis donors. Until 1995, the authors' institution accepted occasional platelet donors with a lower minimum hemoglobin (11.5 g/dL), if accompanied by medical director approval. STUDY DESIGN: All donation records from a 6-month period before 1995 were retrospectively reviewed to determine whether this lower hemoglobin cutoff adversely affected either the safety of the platelet donation process or donors' subsequent hemoglobin levels. RESULTS: Of 450 donations, 56 (12%, Group 1) were from donors with hemoglobin concentrations between 11.5 and 12.4 g per dL (2 donations from 1 man; 54 donations from 45 women). The remaining 394 donations (88%, Group 2) came from donors with hemoglobin concentrations > or = 12.5 g per dL (216 donations from 118 men; 178 donations from 119 women). The frequency of donor reactions was acceptable (Group 1, 11%; Group 2, 6%); 2 percent of donations by Group 1 donors and 1 percent by Group 2 donors were terminated because of these reactions. Of 46 donors in Group 1, 30 returned to donate platelets again at a later time; at least once, 23 (77%) had a hemoglobin > or = 12.5 g per dL. Ten donors in Group 1 returned for additional donations within 56 days; no meaningful decrease in hemoglobin levels occurred. A hemoglobin cutoff of 12.5 g per dL during the study period would have excluded 1 percent of platelet donations by men and 23 percent by women. CONCLUSION: The data demonstrate that the lower hemoglobin cutoff of 11.5 g per dL is a safe and relevant threshold for accepting female plateletpheresis donors and would allow more participation by women in blood donor programs. PMID- 9738627 TI - Comparison of blood component preparation systems based on buffy coat removal: component specifications, efficiency, and process costs. PMID- 9738628 TI - Ethical issues in cord blood banking: summary of a workshop. PMID- 9738629 TI - Monetary blood donation incentives and the risk of transfusion-transmitted infection. PMID- 9738630 TI - Monetary compensation for plasma donors: a record of safety. PMID- 9738631 TI - The Transfusion Alliance: an interdisciplinary approach to improving transfusion medicine. PMID- 9738633 TI - Vascular and lipid syndromes in selected HIV-infected patients. PMID- 9738632 TI - Crossover use of donated blood for autologous transfusion: report of the Council on Scientific Affairs, American Medical Association. AB - BACKGROUND: Controversy exists concerning whether the costs and potential risks outweigh the potential benefits of "crossover" use in the general blood supply of unutilized blood that was donated for autologous transfusion. STUDY DESIGN AND METHODS: Published articles and reports were identified through systematic search of MEDLINE and review of references cited in previously identified articles, textbooks, and reports. Consultation was made with experts in blood donation and transfusion. Additional peer review was received from the American Medical Association (AMA) Council on Scientific Affairs RESULTS: Concern over infectious disease transmission has led to increased interest in and support for autologous transfusion for individuals having planned surgeries. Different requirements exist for collection, labeling, and screening of blood to be used for autologous versus allogeneic transfusions; therefore, procedures for diverting autologous blood donations to the general blood supply involve considerable expense. Several cost-effectiveness studies of autologous blood donation and transfusion conclude that currently this "crossover" appears to be an expensive procedure yielding little increased benefit from a societal perspective. CONCLUSIONS: The recommendations in this report were adopted as AMA Policy at the AMA Annual Meeting in June 1997. The AMA does not encourage blood collection programs to "cross over" units donated for autologous use to the allogeneic blood supply. Practice guidelines are needed, and should be utilized to ensure parsimony in the use of autologous blood donations and transfusions. PMID- 9738634 TI - Withdrawal of Posicor from market. PMID- 9738635 TI - Low-energy atrial defibrillation: a promising new technique. PMID- 9738636 TI - Successful irrigated-tip catheter ablation of atrial flutter resistant to conventional radiofrequency ablation. AB - BACKGROUND: Catheter ablation of typical right atrial flutter is now widely performed. The best end point has been demonstrated to be bidirectional isthmus block. We investigated the use of irrigated-tip catheters in a small subset of patients who failed isthmus ablation with conventional radiofrequency (RF) ablation. METHODS AND RESULTS: Of 170 patients referred for ablation of common atrial flutter, conventional ablation of the cavotricuspid isthmus with >21 applications failed to create a bidirectional block in 13 (7.6%). An irrigated tip catheter ablation was performed on identified gaps in the ablation line according to a protocol found to be safe in animals: a moderate flow rate of 17 mL/min and temperature-controlled (target, 50 degrees C) RF delivery with a power limit of 50 W. Bidirectional isthmus block was achieved in 12 patients by use of a mean delivered power of 40+/-6 W with a single application in 6 patients and 2 to 6 applications in the other 6. No side effects occurred during or after the procedure. CONCLUSIONS: Irrigated-tip catheter ablation is safe and effective for achieving cavotricuspid isthmus block when conventional RF energy has failed. PMID- 9738637 TI - Inflammation, pravastatin, and the risk of coronary events after myocardial infarction in patients with average cholesterol levels. Cholesterol and Recurrent Events (CARE) Investigators. AB - BACKGROUND: We studied whether inflammation after myocardial infarction (MI) is a risk factor for recurrent coronary events and whether randomized treatment with pravastatin reduces that risk. METHODS AND RESULTS: A nested case-control design was used to compare C-reactive protein (CRP) and serum amyloid A (SAA) levels in prerandomization blood samples from 391 participants in the Cholesterol and Recurrent Events (CARE) trial who subsequently developed recurrent nonfatal MI or a fatal coronary event (cases) and from an equal number of age- and sex-matched participants who remained free of these events during follow-up (control subjects). Overall, CRP and SAA were higher among cases than control subjects (for CRP P=0.05; for SAA P=0.006) such that those with levels in the highest quintile had a relative risk (RR) of recurrent events 75% higher than those with levels in the lowest quintile (for CRP RR= 1.77, P=0.02; for SAA RR= 1.74, P=0.02). The study group with the highest risk was that with consistent evidence of inflammation (elevation of both CRP and SAA) who were randomly assigned to placebo (RR=2.81, P=0.007); this risk estimate was greater than the product of the individual risks associated with inflammation or placebo assignment alone. In stratified analyses, the association between inflammation and risk was significant among those randomized to placebo (RR=2.11, P=0.048) but was attenuated and nonsignificant among those randomized to pravastatin (RR=1.29, P=0.5). CONCLUSIONS: Evidence of inflammation after MI is associated with increased risk of recurrent coronary events. Therapy with pravastatin may decrease this risk, an observation consistent with a nonlipid effect of this agent. PMID- 9738638 TI - Helicobacter pylori seropositivity and coronary heart disease incidence. Atherosclerosis Risk In Communities (ARIC) Study Investigators. AB - BACKGROUND: Several epidemiological and clinical reports have suggested seropositivity for Helicobacter pylori may be a risk factor for coronary heart disease. However, there has been no prospective study of this association involving an ethnically diverse sample of middle-aged men and women. METHODS AND RESULTS: Using a prospective, case-cohort design, we determined H pylori seropositivity in relation to coronary heart disease incidence over a median follow-up period of 3.3 years among middle-aged men and women. There were 217 incident coronary heart disease cases and a cohort sample of 498. We determined H pylori antibody status by measuring IgG antibody to the high-molecular-weight cell-associated proteins of H pylori using a sensitive and specific ELISA. The prevalence of H pylori seropositivity was higher in blacks than whites, in those with less than high school education, in those with lower plasma pyridoxal 5' phosphate and higher homocyst(e)ine concentrations, in those who did not use vitamin supplements, in those with higher fibrinogen levels, and in those seropositive for cytomegalovirus and herpes simplex type I (all P<0.05). The age , sex-, race-, and field center-adjusted hazard ratio of coronary heart disease for H pylori seropositivity was 1.03 (95% CI=0.68 to 1.57). After adjustment for other risk factors, including fibrinogen, cytomegalovirus seropositivity, and herpes simplex type I seropositivity, the hazard ratio was 0.85 (95% CI=0.43 to 1.69). H pylori seropositivity also was not associated with increased mean intima media thickness of the carotid artery, a measure of subclinical atherosclerosis. CONCLUSIONS: H pylori infection is probably not an important contributor to clinical coronary heart disease events. PMID- 9738639 TI - Physician noncompliance with the 1993 National Cholesterol Education Program (NCEP-ATPII) guidelines. AB - BACKGROUND: We sought to determine the frequency with which physicians follow National Cholesterol Education Program (NCEP-ATPII) guidelines in screening for cardiovascular risk factors and treating hyperlipidemia. METHODS AND RESULTS: We conducted a retrospective chart review on randomly sampled charts of 225 patients admitted to the coronary care unit between January and June 1996. The main outcome measures were rates of physician screening for coronary heart disease risk factors; rates of counseling for cigarette cessation, diet, and exercise; and extent of use of NCEP algorithms for obtaining LDL cholesterol values and treating hypercholesterolemia. Screening rates for interns (who performed best) were: cigarette use (89%), known coronary heart disease (74%), hypertension (68%), hyperlipidemia (59%), family history (56%), diabetes (37%), postmenopausal hormone therapy (11%), and premature menopause (1%). Four percent of smokers were counseled to quit, 14% of patients were referred to dietitians, and 1% were encouraged to exercise. A full lipid panel was obtained in 50% of patients in whom it was indicated on the basis of NCEP criteria. Patients were more likely to receive lipid-lowering treatment if NCEP criteria indicated that they should, but 36% of hospitalized patients and 46% of patients who should have been treated on discharge were not. CONCLUSIONS: Physicians are poorly compliant with NCEP guidelines for risk factor assessment and counseling, even in patients at high risk for coronary heart disease. Physicians follow NCEP-ATPII algorithms for obtaining an LDL value, a key step in evaluating the need for treatment, only 50% of the time. NCEP criteria seem to influence the decision to initiate lipid lowering therapy, but significant numbers of eligible patients remain untreated. PMID- 9738640 TI - Mechanism of mitral regurgitation in hypertrophic cardiomyopathy: mismatch of posterior to anterior leaflet length and mobility. AB - BACKGROUND: In hypertrophic cardiomyopathy, a spectrum of mitral leaflet abnormalities has been related to the mechanism of mitral systolic anterior motion (SAM), which causes both subaortic obstruction and mitral regurgitation. In the individual patient, SAM and regurgitation vary in parallel; clinically, however, great interindividual differences in mitral regurgitation can occur for comparable degrees of SAM. We hypothesized that these differences relate to variations in posterior leaflet length and mobility, restricting its ability to follow the anterior leaflet (participate in SAM) and coapt effectively. METHODS AND RESULTS: Different mitral geometries produced surgically in porcine valves were studied in vitro. Comparable degrees of SAM resulted in more severe mitral regurgitation for geometries characterized by limited posterior leaflet excursion. Mitral geometry was also analyzed in 23 patients with hypertrophic cardiomyopathy by intraoperative transesophageal echocardiography. All had typical anterior leaflet SAM with significant outflow tract gradients but considerably more variable mitral regurgitation; therefore, regurgitation did not correlate with obstruction. In contrast, mitral regurgitation correlated inversely with the length over which the leaflets coapted (r= -0.89), the most severe regurgitation occurring with a visible gap. Regurgitation increased with increasing mismatch of anterior to posterior leaflet length (r=0.77) and decreasing posterior leaflet mobility (r= -0.79). CONCLUSIONS: SAM produces greater mitral regurgitation if the posterior leaflet is limited in its ability to move anteriorly, participate in SAM, and coapt effectively. This can explain interindividual differences in regurgitation for comparable degrees of SAM. Thus, the spectrum of leaflet length and mobility that affects subaortic obstruction also influences mitral regurgitation in patients with SAM. PMID- 9738641 TI - Assessment of small-diameter aortic mechanical prostheses: physiological relevance of the Doppler gradient, utility of flow augmentation, and limitations of orifice area estimation. AB - BACKGROUND: Noninvasive assessment of functionally stenotic small-diameter aortic mechanical prostheses is complicated by theoretical constraints relating to the hemodynamic relevance of Doppler-derived transprosthetic gradients. To establish the utility of Doppler echocardiography for evaluation of these valves, 20-mm Medtronic Hall and 19-mm St Jude prostheses were studied in vitro and in vivo. METHODS AND RESULTS: Relations between the orifice transprosthetic gradient (equivalent to Doppler), the downstream gradient in the zone of recovered pressure (equivalent to catheter), and fluid mechanical energy losses were examined in vitro. Pressure-flow relations across the 2 prostheses were evaluated by Doppler echocardiography in vivo. For both types of prosthesis in vitro, the orifice was higher than the downstream gradient (P<0.001), and fluid mechanical energy losses were as strongly correlated with orifice as with downstream pressure gradients (r2=0.99 for both). Orifice and downstream gradients were higher and fluid mechanical energy losses were larger for the St Jude than the Medtronic Hall valve (all P<0.001). Whereas estimated effective orifice areas for the 2 valves in vivo were not significantly different, model-independent dynamic analysis of pressure-flow relations revealed higher gradients for the St Jude than the Medtronic Hall valve at a given flow rate (P<0.05). CONCLUSIONS: Even in the presence of significant pressure recovery, the Doppler-derived gradient across small-diameter aortic mechanical prostheses does have hemodynamic relevance insofar as it reflects myocardial energy expenditure. Small differences in function between stenotic aortic mechanical prostheses, undetectable by conventional orifice area estimations, can be identified by dynamic Doppler echocardiographic analysis of pressure-flow relations. PMID- 9738642 TI - Venous levels of shear support neutrophil-platelet adhesion and neutrophil aggregation in blood via P-selectin and beta2-integrin. AB - BACKGROUND: After activation, platelets adhere to neutrophils via P-selectin and beta2-integrin. The molecular mechanisms and adhesion events in whole blood exposed to venous levels of hydrodynamic shear in the absence of exogenous activation remain unknown. METHODS AND RESULTS: Whole blood was sheared at approximately 100 s(-1). The kinetics of neutrophil-platelet adhesion and neutrophil aggregation were measured in real time by flow cytometry. P-selectin was upregulated to the platelet surface in response to shear and was the primary factor mediating neutrophil-platelet adhesion. The extent of neutrophil aggregation increased linearly with platelet adhesion to neutrophils. Blocking either P-selectin, its glycoprotein ligand PSGL-1, or both simultaneously by preincubation with a monoclonal antibody resulted in equivalent inhibition of neutrophil-platelet adhesion (approximately 30%) and neutrophil aggregation (approximately 70%). The residual amount of neutrophil adhesion was blocked with anti-CD11b/CD18. Treatment of blood with prostacyclin analogue ZK36374, which raises cAMP levels in platelets, blocked P-selectin upregulation and neutrophil aggregation to baseline. Complete abrogation of platelet-neutrophil adhesion required both ZK36374 and anti-CD18. Electron microscopic observations of fixed blood specimens revealed that platelets augmented neutrophil aggregation both by forming bridges between neutrophils and through contact-mediated activation. CONCLUSIONS: The results are consistent with a model in which venous levels of shear support platelet adherence to neutrophils via P-selectin binding PSGL-1. This interaction alone is sufficient to mediate neutrophil aggregation. Abrogation of platelet adhesion and aggregation requires blocking Mac-1 in addition to PSGL-1 or P-selectin. The described mechanisms are likely of key importance in the pathogenesis and progression of thrombotic disorders that are exacerbated by leukocyte-platelet aggregation. PMID- 9738643 TI - Low-energy cardioversion with epicardial wire electrodes: new treatment of atrial fibrillation after open heart surgery. AB - BACKGROUND: Atrial fibrillation (AF) is the most common arrhythmia after open heart surgery. Traditional treatment with a range of antiarrhythmic drugs and electrical cardioversion is associated with considerable side effects. The aim of this study was to examine the feasibility and efficacy of low-energy atrial defibrillation with temporary epicardial defibrillation wire electrodes. METHODS AND RESULTS: Epicardial defibrillation wire electrodes were placed at the left and right atria during open heart surgery in 100 consecutive patients (age 65+/-9 years; male to female ratio 67:23). Electrophysiological studies performed postoperatively revealed a test shock (0.3 J) impedance of 96+/-12 omega (monophasic) and 97+/-13 omega (biphasic). During their hospital stay, AF occurred in 23 patients (23%) at 2.1+/-1.3 days postoperatively. Internal atrial defibrillation was performed in 20 patients. Of these patients, 80% (16/20) were successfully cardioverted with a mean energy of 5.2+/-3 J. Early recurrence of AF (< or =60 seconds after defibrillation) developed in 8 patients. Five patients had multiple episodes of AF. In total, 35 episodes of AF were treated, with an 88% success rate. Only 6 patients (30%) required sedation. No complications were observed with shock application or with lead extraction. CONCLUSIONS: Atrial defibrillation with temporary epicardial wire electrodes can be performed safely and effectively in patients after cardiac operations. The shock energy required to restore sinus rhythm is low. Thus, patients can be cardioverted without anesthesia. PMID- 9738644 TI - Simultaneous endocardial mapping in the human left ventricle using a noncontact catheter: comparison of contact and reconstructed electrograms during sinus rhythm. AB - BACKGROUND: Catheter ablation of ventricular tachycardia is limited in part by difficulty in identifying suitable sites for ablation. A noncontact multielectrode array (MEA) has been developed that allows reconstruction of 3360 electrograms, using inverse-solution mathematics, that are superimposed onto a computer-simulated model of the endocardium. This study assesses the accuracy of timing and morphology of reconstructed unipolar electrograms compared with contact unipolar electrograms from the same endocardial site. METHODS AND RESULTS: The MEA was deployed in the left ventricles of 13 patients (end diastolic diameters, 61.7+/-8.4 mm [mean+/-SD]). We recorded contact electrograms at 76 points equatorial and 32 points nonequatorial to the MEA during sinus rhythm using a catheter-locator signal to record direction and distance from the MEA. Morphology (cross-correlation) and timing of maximum -dV/dt of contact and reconstructed electrograms were compared at different distances from the MEA center to endocardium (M-E) and from the MEA equatorial plane. For equatorial data, the M-E was 32.12+/-12.12 mm. The timing of reconstructed with respect to contact electrograms was -1.94+/-7.12 ms for M-E <34 mm and -14.16+/-19.29 ms at M-E >34 mm (P<0.001). Cross-correlation of electrograms was 0.87+/-0.12 (95% CI, 0.84 to 0.91) and 0.76+/-0.18 (95% CI, 0.69 to 0.83) for M-E <34 mm and >34 mm, respectively. Nonequatorial points were 32.33+/-10.81 mm (range, 16.9 to 55.6 mm) from the MEA equatorial plane; electrogram timing difference was -8.97+/-15.75 ms and was unrelated to this distance from the equator. CONCLUSIONS: This noncontact mapping system accurately reconstructs endocardial unipolar electrograms from the human left ventricle. At M-E distances >34 mm, timing accuracy of reconstruction decreases. PMID- 9738645 TI - Fostriecin, an inhibitor of protein phosphatase 2A, limits myocardial infarct size even when administered after onset of ischemia. AB - BACKGROUND: The role of protein phosphatases (PPs) during ischemic preconditioning in the rabbit heart was examined. METHODS AND RESULTS: Fostriecin, a potent inhibitor of PP2A, was administered to isolated rabbit hearts starting either 15 minutes before or 10 minutes after the onset of a 30 minute period of regional ischemia and continuing until the onset of reperfusion. After 2 hours of reperfusion, infarct size was measured with triphenyltetrazolium chloride. In a second study with isolated rabbit cardiomyocytes, the effect of fostriecin pretreatment was assessed by measuring changes in cell osmotic fragility during simulated ischemia. PP1 and PP2A activities of isolated control and ischemically preconditioned cells were also measured. In a third series of experiments, left ventricular biopsies of isolated rabbit hearts were obtained before and at selected times during 60 minutes of global ischemia, and the tissue was assayed for PP1 and PP2A activities. In isolated hearts pretreated with fostriecin, only 8% of the ischemic zone infarcted, significantly less than that in untreated control hearts (33%; P<0.001) but comparable to that in ischemically preconditioned hearts (9%; P<0.001 versus control). Significant protection was also observed in the hearts treated only after the onset of ischemia (18% infarction; P<0.05 versus control). In isolated myocytes, fostriecin also provided protection comparable to that produced by metabolic preconditioning. Preconditioning had no apparent effect on the activity of either PP1 or PP2A in isolated ventricular myocytes or ventricular tissue obtained from heart biopsies. CONCLUSIONS: Fostriecin, a potent inhibitor of PP2A, can protect the rabbit heart from infarction even when administered after the onset of ischemia. But inhibition of either PP1 or PP2A does not appear to be the mechanism of protection from ischemic preconditioning. PMID- 9738646 TI - Enhancement of monocyte procoagulant activity by adhesion on vascular smooth muscle cells and intercellular adhesion molecule-1-transfected Chinese hamster ovary cells. AB - BACKGROUND: Plaque erosion is a frequent finding in sudden death due to coronary thrombosis. The present study sought to investigate whether monocyte adhesion to human aortic vascular smooth muscle cells (VSMCs) induces procoagulant activity (PCA) and whether this could be mediated by intercellular adhesion molecule-1 (ICAM-1). METHODS AND RESULTS: We incubated mononuclear cells (MNCs) with VSMCs and ICAM-1-transfected Chinese hamster ovary (CHO) cells, investigated monocyte tissue factor (TF) mRNA expression by Northern blot analysis and TF protein expression by ELISA, and measured PCA. Incubation of MNCs with VSMCs for 6 hours increased PCA from 0.7+/-0.1 to 166.0+/-37.9 mU/105 cells (P=0.007), which could be inhibited in a dose-dependent manner by the addition of blocking anti-ICAM-1 monoclonal antibodies. Prestimulation of VSMCs with interleukin-1beta enhanced surface ICAM-1 expression significantly but did not induce PCA in VSMCs. Incubation of MNCs with prestimulated VSMCs led to a further increase in PCA to 239.9+/-27.9 mU/10(5) cells (P=0.02 compared with incubation with unstimulated VSMCs). Incubation of MNCs with VSMCs enhanced TF mRNA after 2 hours and significantly increased TF protein content after 6 hours. Incubation of purified monocytes with ICAM-1-transfected CHO cells increased PCA from 1.2+/-0.2 to 81.9+/-3.3 mU/10(5) cells (P<0.001 compared with incubation with untransfected CHO cells) after 6 hours. This effect could be inhibited significantly by the addition of blocking anti-CD18, anti-CD11b, or anti-CD11c monoclonal antibodies. Similar results were obtained for MNCs. CONCLUSIONS: Monocyte adhesion to VSMCs induces TF mRNA and protein expression and monocyte PCA, which is regulated by beta2-integrin-mediated monocyte adhesion to ICAM-1 on VSMCs. PMID- 9738647 TI - Noninvasive, in utero imaging of mouse embryonic heart development with 40-MHz echocardiography. AB - BACKGROUND: The increasing number of transgenic and targeted mutant mice with embryonic cardiac defects has resulted in the need for noninvasive techniques to examine cardiac structure and function in early mouse embryos. We report the first use of a novel 40-MHz ultrasound imaging system in the study of mouse cardiac development in utero. METHODS AND RESULTS: Transabdominal scans of mouse embryos staged between 8.5 and 13.5 days of gestation (E8.5 to E13.5) were obtained in anesthetized mice. Atrial and ventricular contractions could be discerned from E9.5, and changes in cardiac morphology were observed from E9.5 to E13.5. Hyperechoic streaming patterns delineated flow through the umbilical, vitelline, and other major blood vessels. Diastolic and systolic ventricular areas were determined by planimetry of the epicardial borders, and fractional area change was measured as an index of contractile function. Significant increases in ventricular size were documented at each stage between E10.5 and E13.5, and the ability to perform serial imaging studies over 3 days of embryonic development is described. Finally, the detection of vascular cell adhesion molecule 1 (VCAM-1) homozygous null mutant embryos demonstrates the first example of noninvasive, in utero analysis of cardiac structure and function in a targeted mouse mutant. CONCLUSIONS: We used 40-MHz echocardiography to identify key elements of the early mouse embryonic cardiovascular system and for noninvasive dimensional analysis of developing cardiac ventricles. The ability to perform serial measurements and to detect mutant embryos with cardiac defects highlights the usefulness of the technique for investigating normal and abnormal cardiovascular development. PMID- 9738648 TI - Local adenovirus-mediated transfer of human endothelial nitric oxide synthase reduces luminal narrowing after coronary angioplasty in pigs. AB - BACKGROUND: Nitric oxide, synthesized from L-arginine by nitric oxide synthase (NOS), is a vasodilator and inhibits vascular smooth muscle cell (SMC) proliferation and migration. The effects of local NOS gene transfer on restenosis after experimental balloon angioplasty were investigated. METHODS AND RESULTS: Left anterior descending coronary artery angioplasty was performed in 25 pigs. Animals received an intramural injection of adenovirus (1.5 x 10(9) pfu) carrying either the NOS cDNA (AdCMVceNOS) or no cDNA (AdRR5) via the Infiltrator. Local gene transfer efficiency and bioactivity of recombinant protein were assessed after 4 days. Indices of restenosis were evaluated by computerized planimetry on coronary artery sections prepared 28 days after angioplasty. Adenoviral vectors permitted efficient gene delivery to medial SMCs and adventitial cells of coronary arteries. Vascular cGMP levels were depressed after angioplasty from 1.30+/-0.42 to 0.33+/-0.20 pmol/mg protein (P<0.05) but were restored after constitutive endothelial (ce) NOS gene transfer to 1.82+/-0.98 pmol/mg (P<0.05 versus injured group and P=NS versus control). The ratio of the neointimal area to the internal elastic lamina fracture length, maximal neointimal thickness, and percent stenosis were all reduced in AdCMVceNOS- versus AdRR5-transduced pigs (0.59+/-0.14 versus 0.80+/-0.19 mm, P=0.02; 0.75+/-0.21 versus 1.04+/-0.25 mm, P=0.019; and 53+/-15% versus 75+/-11%, P=0.006, respectively). Lumen area was significantly larger (0.70+/-0.35 mm2 in AdCMVceNOS versus 0.32+/-0.18 mm2 in AdRR5, P=0.007). CONCLUSIONS: Percutaneous adenovirus-mediated NOS gene transfer resulted in efficient local overexpression of functional NOS after angioplasty in coronary arteries. Restored NO production in injured coronary arteries significantly reduced luminal narrowing, most likely through a combined effect on neointima formation and on vessel remodeling after angioplasty. PMID- 9738649 TI - Stent-supported carotid angioplasty: should it be done, and, if so, by whom? A 1998 perspective. PMID- 9738650 TI - Images in cardiovascular medicine. Mobile left atrial thrombus associated with mitral stenosis. PMID- 9738651 TI - Images in cardiovascular medicine. Left ventricular thrombus detected by two- and three-dimensional computed tomographic ventriculography: a new application of helical CT. PMID- 9738652 TI - Very low fat diets. PMID- 9738653 TI - Tumor-induced immune dysfunction: the macrophage connection. AB - Although macrophages (Mphis) mediate tumor cytotoxicity, display tumor-associated antigens, and stimulate antitumor lymphocytes, cancer cells routinely circumvent these host-mediated immune activities, rendering the host incapable of mounting a successful antitumor immune response. Evidence supporting a direct causal relationship between cancer and immune dysfunction suggests that the presence of neoplastic tissue leads to immunologic degeneration. Furthermore, substantial data demonstrate that tumor growth adversely alters Mphi function and phenotype. Thus, although Mphis can serve as both positive and negative mediators of the immune system, the importance of Mphis in tumor-induced immune suppression remains controversial. This review focuses on the evidence that tumor-derived molecules redirect Mphi activities to promote tumor development. Tumors produce cytokines, growth factors, chemotactic molecules, and proteases that influence Mphi functions. Many tumor-derived molecules, such as IL-4, IL-6, IL-10, MDF, TGF beta1, PGE2, and M-CSF, deactivate or suppress the cytotoxic activity of activated Mphis. Evidence that tumor-derived molecules modulate Mphi cytotoxicity and induce Mphi suppressor activity is presented. This information further suggests that Mphis in different in vivo compartments may be differentially regulated by tumor-derived molecules, which may deactivate tumor-proximal (in situ) Mphi populations while concurrently activating tumor-distal Mphis, imparting a twofold insult to the host's antitumor immune response. PMID- 9738654 TI - Combinatorial requirements for adhesion molecules in mediating neutrophil emigration during bacterial peritonitis in mice. AB - To investigate the requirements for adhesion molecules in neutrophil emigration during peritonitis, mice received intraperitoneal injections of Streptococcus pneumoniae while the functions of multiple adhesion molecules were blocked. Emigration after 4 h was compromised by antibodies against ICAM-1 or genetic deficiency of ICAM-1. Anti-CD11a/CD18 antibodies decreased emigration in ICAM-1 mutant mice, suggesting that ICAM-1 independent emigration requires CD11/CD18 complexes. In contrast, mice mutant in ICAM-1 plus E-selectin showed no defect in emigration, suggesting that E-selectin commits neutrophils to an ICAM-1-dependent pathway during streptococcal peritonitis. However, in mutant mice lacking the three endothelial adhesion molecules E-selectin, P-selectin, and ICAM-1, emigration after 4 h was significantly compromised. Thus, P-selectin is essential to ICAM-1- and E-selectin-independent acute peritoneal inflammation. After 24 h of peritonitis, there were no differences between WT and E-selectin/P selectin/ICAM-1 mutant mice, demonstrating that these endothelial adhesion molecules are not essential to neutrophil emigration during later stages of peritonitis. PMID- 9738655 TI - HIV load in highly purified CD8+ T cells retrieved from pulmonary and blood compartments. AB - The pulmonary microenvironment can be HIV infected from the asymptomatic period of HIV infection and it has been demonstrated that HIV presence elicits a discrete virus-specific CTL immune response in the lungs. In this study we analyzed T cell subsets and compared the proportion of CD8+ T cells harboring HIV in the lung and in the circulation of HIV-seropositive patients with T cell alveolitis. Proviral load was assessed using the DNA-polymerase chain reaction technique on highly purified CD8+ T cells isolated from peripheral blood and bronchoalveolar lavage. The proviral load of pulmonary CD8+ T cells showed an upward trend with respect to corresponding samples isolated from the peripheral blood of the same patients. PMID- 9738656 TI - Macrophage cytoplasmic vesicle pH gradients and vacuolar H+-ATPase activities relative to virus infection. AB - A number of viruses replicate in macrophages, some having an obligate requirement for a macrophage host. This raised the question concerning the role of the macrophage endosomal/lysosomal compartment during such infections. Both lysosomotropic weak bases, amantadine and chloroquine, which interfere with endosomal/lysosomal pH gradients, and the macrolide antibiotic bafilomycin A1, which interferes with vacuolar H+-ATPase, inhibited African swine fever (ASF) virus replication in porcine macrophages. This inhibition was reversible: replenishment of bafilomycin, but not amantadine or chloroquine, maintained the inhibition. The characteristics of the inhibition, and the capacity of virus to escape and re-commence replication, related to the capacity of each drug to interfere with the endosomal/lysosomal proton pump. These results demonstrate that the virus actually requires macrophage endosomal/lysosomal activity for its replication. Therein, vacuolar H+-ATPase activity is particularly critical for successful virus replication, which is interesting considering the importance of this for endosomal/lysosomal activity and macrophage function. PMID- 9738657 TI - Gangliosides of monocyte-derived macrophages of adults with advanced HIV infection show reduced surface accessibility. AB - Gangliosides of macrophages are potent immunoregulatory molecules. A monoclonal antibody directed at human macrophage gangliosides (25F4) inhibits macrophage migration with relative specificity. Recent reports suggested that greater expression of G(M1) in mononuclear cells accompanies advanced HIV infection, although others failed to demonstrate any differences in vitro. We purified gangliosides from blood monocyte-derived macrophages obtained from HIV-infected adults. Densitometric analysis of chromatograms demonstrated no differences in relative quantities of any macrophage gangliosides among all HIV-positive and negative donors. Antibody 25F4 showed equivalent ELISA reactivity with purified macrophage gangliosides of HIV-positive and -negative donors. However, intact macrophages of HIV donors with CD4+ cell counts <200/mm3 showed impaired immunofluorescent surface expression of the 25F4 epitope and concomitant loss of migration inhibitory responsiveness. Thus, although relative content is unchanged, macrophage gangliosides become surface-inaccessible in adults with advanced HIV infection. Our data provide further evidence that dysregulation of glycosphingolipid metabolism in HIV-1 infection contributes to immune dysfunction. PMID- 9738658 TI - Effect of trifluoromethyl ketone-based elastase inhibitors on neutrophil function in vitro. AB - Neutrophils release elastase, which is known secondarily to cause tissue damage. However, it is rapidly inactivated by the endogenous alpha1-proteinase inhibitor (alpha1Pi). Nevertheless, under pathological conditions, alpha1i is inactivated by oxidants released from neutrophils, resulting in an excess of elastase at the site of inflammation. This elastase/alpha1Pi imbalance has been implicated as a pathogenic factor in cystic fibrosis, acute respiratory distress syndrome, and emphysema. Elastase inhibitors, which do not interfere with the microbicidal activity of neutrophils and are resistant to neutrophil-released oxidants, would undoubtedly represent an important advance in the management of neutrophil mediated tissue injury. We report that a new family of elastase inhibitors ICI200355 and ZD0892 was found to be resistant toward superoxide, hypochlorous acid, hydrogen peroxide, hydroxyl radical, and peroxynitrite mediated degradation as well as having no effect on the formation of these oxidants by activated neutrophils. More importantly, we found that these inhibitors did not interfere with the ability of human neutrophils to phagocytose and to kill Staphylococcus aureus. In conclusion, a new potent class of elastase inhibitors, while blocking the effects of neutrophil elastase, was found not to impede various physiological functions of human neutrophils, in particular the ability of these phagocytic cells to phagocytose and kill bacteria. PMID- 9738659 TI - Spontaneous and Fas-mediated apoptosis are diminished in umbilical cord blood neutrophils compared with adult neutrophils. AB - Apoptosis mediates neutrophil (PMN) phagocytosis and is influenced by cytokines and the Fas/Fas ligand pathway. To determine whether apoptosis of cord blood PMN differs from those of adults, cultured PMN were evaluated by morphological analysis, flow cytometry (TUNEL assay), and DNA gel electrophoresis. In addition, we studied the effect of anti-Fas IgM or cycloheximide on induction of PMN apoptosis. Spontaneous apoptosis (24 h) was less in cord blood PMN (mean +/- SD; 29 +/- 9 vs. adults, 56 +/- 14%, P < 0.001). Treatment (6 h) with anti-Fas IgM induced less apoptosis in cord blood PMN (24 +/- 6 vs. adults, 63 +/- 7%, P < 0.001), as did treatment with cycloheximide (13 +/- 10 vs. adults, 55 +/- 16%, P < 0.01). These data suggest the pre-existence of proteins that inhibit apoptosis or the absence of those that promote apoptosis in cord blood PMN. PMID- 9738661 TI - Characterization of a rat alveolar macrophage cell line that expresses a functional mannose receptor. AB - The mannose receptor is a single polypeptide transmembrane glycoprotein expressed on the surface of macrophages that binds and internalizes soluble and particulate ligands. Physiological ligands for this receptor are pathogens, such as mycobacteria, and extracellular acid hydrolases and peroxidases. Expression of the mannose receptor is tightly linked to the functional state of the macrophage: the receptor appears during differentiation, is increased by macrophage deactivating agents, and is reduced in the presence of macrophage activating agents. Studies of the mechanisms underlying these regulatory processes have been hampered by the lack of a stable cell line that expresses a functional and appropriately regulated mannose receptor. In this study we describe expression and modulation of the mannose receptor by the rat alveolar cell line NR8383. Similar amounts of the mannose receptor ligand horseradish peroxidase were internalized by both NR8383 cells and alveolar macrophages. In addition, NR8383 cells expressed immunoreactive mannose receptor protein and mannose receptor mRNA as detected by Northern analysis. Regulation studies showed that mannose receptor expression was regulated at the levels of activity, protein, and mRNA in NR8383 cells similarly to regulation in primary rat macrophages. In addition, NR8383 cells could be successfully transfected with a luciferase reporter gene, providing the transfectable, mannose receptor-positive macrophage cell line. These results support the hypothesis that NR8383 cells potentially represent the best current macrophage cell line for studying various aspects of macrophage function, and are particularly critical in studies of regulation of the mannose receptor, a key receptor in host defense and immune regulation. PMID- 9738660 TI - Activation of rat macrophages by Betafectin PGG-glucan requires cross-linking of membrane receptors distinct from complement receptor three (CR3). AB - PGG-glucan (Betafectin) is a soluble, highly purified yeast (1,3)-beta-glucan with broad anti-infective and immunomodulatory activities. These studies evaluated the ability of PGG-glucan to directly elicit O2- and tumor necrosis factor alpha (TNF-alpha) production by rat leukocytes in vitro. Particulate beta glucan stimulated O2- production by the rat NR8383 alveolar macrophage cell line and resident rat peritoneal macrophages, but soluble PGG-glucan did not. In contrast, presentation of PGG-glucan to cells after covalent immobilization to a plastic surface caused a direct stimulation of O2- and TNF-alpha production. The O2- response of rat leukocytes to immobilized PGG-glucan was inhibited by soluble PGG-glucan, indicating that cellular responses to both immobilized and soluble PGG-glucan occur via common cell surface receptors. Because complement receptor type three (CR3) has been proposed as a beta-glucan receptor on human leukocytes, NR8383 cells were evaluated for the presence of CR3. Indirect immunofluorescence and flow cytometric analysis showed that despite being responsive to both particulate and immobilized beta-glucans, NR8383 cells expressed no detectable CR3. These results indicate that the beta-glucan receptors on NR8383 cells are not CR3 and suggest that physical presentation plays an important role in inducing pro-inflammatory leukocyte responses to PGG-glucan. PMID- 9738662 TI - A 70-kDa amino-terminal fibronectin fragment supports gelatin binding to macrophages and decreases gelatinase activity. AB - We previously reported that a macrophage response that increased binding to 125I radiolabeled soluble denatured collagen (gelatin) was induced by preincubation of macrophage with a 70-kDa amino-terminal fibronectin fragment and soluble nonlabeled gelatin [S. F. Penc, F. A. Blumenstock, J. E. Kaplan (1995) J. Leukoc. Biol. 58, 501-509]. We now report that neither protein synthesis nor recycling of receptors between the cell surface and interior were required for this response. However, removal of cell surface components with trypsin demonstrated that induced gelatin binding required native cell surface constituents. It was found that in the presence of the 70-kDa fibronectin fragment and gelatin, matrix metalloprotease-2 (MMP-2) and matrix metalloprotease-9 (MMP-9) activity in the cell layers was significantly decreased or undetectable, respectively. Similar levels of increased gelatin binding could be reproduced after inhibition of matrix-degrading metalloprotease activity with 1'10-phenanthroline. These results demonstrate that a macrophage specific response that decreased gelatinase activity and increased gelatin binding was initiated by interaction with a 70-kDa fibronectin fragment and gelatin. PMID- 9738663 TI - Interferon-alpha and granulocyte-macrophage colony-stimulating factor differentiate peripheral blood monocytes into potent antigen-presenting cells. AB - The diverse roles of interferon-alpha (IFN-alpha) in regulating the immune response to infectious agents suggested that it might affect dendritic cell (DC) development. Peripheral blood mononuclear cells cultured with IFN-alpha and granulocyte-macrophage colony-stimulating factor (GM-CSF) developed a dendritic morphology and expressed high levels of the class I and II human leukocyte antigens (HLA), B7 co-stimulatory molecules, adhesion proteins, and CD40. Elevated DC expression of B7-2 and HLA-DR was observed with increasing IFN-alpha concentrations up to 5000 U/mL. The effects of IFN-alpha on DC immunophenotype were not reversed by adding neutralizing antibodies against interleukin-4 (IL-4) or tumor necrosis factor alpha to the cell cultures or by eliminating lymphocytes from the cultures. The addition of IFN-alpha to cultures containing optimal concentrations of IL-4 and GM-CSF significantly increased the B7-2 and HLA-DR levels above those present on DCs grown in two cytokines. The DCs generated with IFN-alpha and GM-CSF were potent antigen-presenting cells in allogeneic mixed leukocyte reactions. They also were capable of taking up, processing, and presenting tetanus toxin to autologous T lymphocytes. These results demonstrate an important role for IFN-alpha in the generation of DCs with potent antigen presenting capabilities from peripheral blood monocytes. PMID- 9738665 TI - Fas ligand (gld)- and Fas (lpr)-deficient mice do not show alterations in the extravasation or apoptosis of inflammatory neutrophils. AB - Apoptosis of neutrophils plays a critical role in the resolution of acute inflammation. Neutrophils from human peripheral blood express Fas (CD95) and are sensitive to Fas ligand (FasL)/Fas-mediated apoptosis. Mice carrying spontaneous mutations in the genes for fas ligand (B6/gld) or fas (B6/lpr) were used to assess the role of FasL/Fas in the kinetics and magnitude of neutrophil extravasation to the thioglycolate (TG)-inflamed peritoneum and in the spontaneous apoptosis of TG-elicited neutrophils. The results showed that TG elicited neutrophils (defined by flow cytometry as GR-1/Ly-6G(hi) cells) from normal (B6) and B6/gld mice, but not from the Fas-deficient B6/lpr mice, express high levels of Fas. The TG-elicited neutrophil response began at 2 h, peaked at 4 h, and subsided by 24-48 h after TG administration in all three strains. However, the response was more prolonged in B6 mice, such that B6/gld and B6/lpr mice had fewer neutrophils at 6 h after TG administration than did B6 mice. Further studies showed that 4 h TG-elicited neutrophils from B6, B6/gld and B6/lpr mice undergo apoptosis in vitro at similar rates (as assessed through flow cytometry by the decrease in forward angle light-scatter and externalization of phosphatidylserine (PS; as detected by Annexin V-FITC) that occur as neutrophils undergo apoptosis). Fas expression was down-regulated on apoptotic neutrophils in conjunction with maximal PS externalization and decreased forward angle light scatter. Collectively, these findings suggest that FasL/Fas-mediated apoptosis is not essential in regulating the lifespan of neutrophils during an acute inflammatory response. The abbreviated inflammatory response observed in FasL/Fas deficient mice is likely to be a secondary effect of the gld/lpr autoimmune/lymphoproliferative syndrome, and not a direct effect of FasL/Fas on the ability of inflammatory neutrophils to undergo apoptosis. PMID- 9738664 TI - Lipopolysaccharide-stimulated TNF-alpha release from cultured rat Kupffer cells: sequence of intracellular signaling pathways. AB - We recently hypothesized that lipopolysaccharide (LPS) stimulation of rat Kupffer cells to induce tumor necrosis factor alpha (TNF-alpha) release requires internalization of LPS, acidification of endosomes, elevation of intracellular calcium, protein kinase C (PKC) activation, and protein tyrosine kinase (PTK) activation. This study uses inhibitors in pulse-chase experiments to determine the sequence of events of intracellular signals required for LPS-stimulated TNF alpha release from Kupffer cells. Inhibitors of internalization (cytochalasin B, monodansylcadaverine) prevented LPS-stimulated TNF-alpha release when added simultaneously with LPS but when added 10 min after LPS, no significant inhibition occurred. The inhibitor of PTK, tyrphostin AG, blocked TNF-alpha release by only 39 +/- 4% (P < 0.001 compared with TNF-alpha release when added simultaneously with LPS) when added 10 min after LPS. Inhibitors of endosomal acidification (bafilomycin A, monensin) inhibited LPS-stimulated TNF-alpha release by 92 +/- 11% (P < 0.001 when no inhibitor was used) when added 10 min after LPS and their effect was totally abrogated when added 45 min after LPS. The PKC inhibitor, H-7, blocked TNF-alpha release by 94 +/- 9% (P < 0.001 when no inhibitor was used) when added 30 min after LPS. The calcium channel blocker, nisoldipine, still inhibited LPS-stimulated TNF-alpha release when added 45 min after LPS. These data support the hypothesis that for LPS-stimulated TNF-alpha release in Kupffer cells, LPS must first be internalized, which may stimulate PTK activation. An intermediate step of signaling involves endosomal acidification. Elevation of intracellular calcium and PKC activation occur as late intracellular signaling events. PMID- 9738666 TI - Contribution of the CXC chemokines IP-10 and Mig to the antitumor effects of IL 12. AB - The mechanisms by which interleukin-12 (IL-12) exerts antitumor effects have been difficult to dissect. In this study, we examined the potential contribution of the chemokines interferon-gamma-inducible protein-10 (IP-10) and Mig to the antitumor effects of IL-12. Using an athymic mouse model, local inoculations with IL-12 consistently produced tumor size reductions associated with characteristic tumor necrosis and vascular damage. These effects were indistinguishable from those produced by IP-10 or Mig injected locally in the same tumor model. Local and systemic treatment with IL-12 was associated with expression of the interferon-gamma (IFN-gamma), IP-10, and Mig genes and proteins in the tumor. Levels of IP-10 and Mig expression in the tumor, the liver, and the kidney were inversely correlated with tumor size. Administration in vivo of neutralizing antibodies to IP-10 and Mig reduced substantially the antitumor effects of IL-12 inoculated locally into the tumors. These results support the notion that IP-10 and Mig contribute to the antitumor effects of IL-12 through their inhibitory effects on tumor vasculature. PMID- 9738667 TI - Interferon-gamma gene expression in cycling and pregnant mouse uterus: temporal aspects and cellular localization. AB - Interferon-gamma (IFN-gamma) is a potent pro-inflammatory cytokine that modulates hematopoietic cell maturation, differentiation, activation, and apoptosis. To evaluate the postulate that locally produced IFN-gamma could influence uterine hematopoietic cells, specific protein was detected by immunohistochemistry and messenger RNA (mRNA) was identified by in situ hybridization in cycling and pregnant mouse uteri. In cycling uteri, IFN-gamma was limited to luminal and glandular epithelial cells during the estrus phase of the cycle. In pregnant uteri, IFN-gamma was prominent at early (gestation day 6-10) and late (gestation day 18) stages. IFN-gamma-producing cells identified by in situ hybridization included uterine epithelial cells, natural killer cells, macrophages, placental trophoblast cells, and cells in the degenerating metrial gland. Collectively, the data indicate that programming of immune and other cells via autocrine and paracrine pathways could be achieved by locally produced IFN-gamma, and suggest that uteroplacental IFN-gamma may be most influential during early and late stages of pregnancy. PMID- 9738668 TI - Antibody-induced engagement of beta2 integrins in human neutrophils causes a rapid redistribution of cytoskeletal proteins, Src-family tyrosine kinases, and p72syk that precedes de novo actin polymerization. AB - Beta2 integrins mediate rearrangement of the cytoskeleton and activation of selective cell functions in neutrophils. To elucidate early events following beta2 integrin ligation, we analyzed redistribution of cytoskeletal and signaling proteins as a consequence of antibody-induced integrin clustering. Incubation of neutrophils on surface-bound anti-beta2 subunit antibodies induced a very rapid (within 1 min) redistribution of the cytoskeletal proteins talin, alpha-actinin, and paxillin, and the tyrosine kinases p58(c-fgr), p53/56(lyn), and p72(syk) to a cell fraction insoluble in Triton X-100. Cytoskeletal and signaling proteins redistribution preceded de novo actin polymerization because cytochalasin B did not inhibit this redistribution. Antibody engagement of all the three distinct beta2 integrins (CD11a, CD11b, CD11c) expressed by neutrophils induced redistribution of cytoskeletal proteins and tyrosine kinases. Several tyrosine phosphorylated proteins were also rapidly redistributed as a consequence of beta2 integrin engagement and this was not affected by blocking de novo actin polymerization with cytochalasin B. In addition, genistein, an inhibitor of tyrosine kinase activities which strongly reduced protein tyrosine phosphorylation, had no effect on redistribution of cytoskeletal proteins, Src family kinases, and p72(syk). These findings suggest that integrin-dependent cytoskeleton rearrangement in neutrophils occurs in at least two distinct steps and nucleation of some cytoskeletal proteins together with tyrosine kinases precedes rearrangement of the actin-based cytoskeleton and tyrosine kinases activation. On the basis of these and previous findings we propose a model explaining mechanisms of integrin signaling in neutrophils. PMID- 9738669 TI - SB202190, a selective inhibitor of p38 mitogen-activated protein kinase, is a powerful regulator of LPS-induced mRNAs in monocytes. AB - Inhibitors of p38 mitogen-activated protein kinase (p38) have been reported to block tumor necrosis factor alpha (TNF-alpha) and interleukin-1beta (IL-1beta) production in monocytes at the level of mRNA translation. Yet, several studies document that p38 can phosphorylate and activate specific transcription factors. Thus, to understand better the role of p38 during monocyte activation, we sought to determine the extent to which p38 is required for lipopolysaccharide (LPS) induced gene expression. For this, differential mRNA display was used to identify LPS-induced genes whose expression was blocked by SB202190, a specific inhibitor of p38. A partial screen identified 10 genes in monoyctes induced 4- to 74-fold by LPS. Of these, genes encoding interferon-induced gene 15, neuroleukin, radiation-inducible immediate-early gene-1, A20, IL-1beta, and superoxide dismutase were suppressed >50% by SB202190. LPS-induced gene activation was not blocked by cycloheximide, indicating that synthesis of intermediate proteins was not required. SB202190 blocked gene induction by 50% when present between 41 and 123 nM, consistent with the potency of this compound as a p38 inhibitor. Furthermore, the ability of SB202190 to block gene activation was stimulus dependent. LPS and interferon-alpha (IFN-alpha) both up-regulated neuroleukin mRNA, but only LPS-induced neuroleukin mRNA was suppressed by SB202190. In contrast, TNF-alpha and LPS both induced IL-8 mRNA, and induction by either TNF alpha or LPS was blocked by SB202190. These data were consistent with the ability of LPS and TNF-alpha, but not IFN-alpha, to activate p38 in monocytes. The results provide pharmacological evidence that p38 may be a key mediator of inducible gene expression in monocytes, but its role is stimulus and gene specific. PMID- 9738670 TI - Antiepileptic drugs, learning, and behavior in childhood epilepsy. AB - Cognitive and behavioral impairments are found more often among epileptic children than among their peers. The cause of these impairments is multifactorial. Identifying the relative contribution of antiepileptic drugs (AEDs) to these problems has been the object of a large number of clinical investigations. This area of research has been characterized by an unusually high number of methodological challenges and pitfalls. Accordingly, results have often been inconsistent and contradictory, except for the more obvious observations that can be derived from clinical experience. Overall, the effects of AEDs on cognition and behavior in children have been overrated in the past. More recent research has benefited from the methodological lessons of previous studies and it suggests that the majority of children taking AEDs do not experience clinically relevant cognitive of behavioral adverse effects from these medications. In addition, some of the newer AEDs may indeed have a better cognitive profile. Nevertheless, clinical experience must be used to identify the subgroup of children who remain at risk for significant and clinically relevant cognitive and behavioral adverse effects of AEDs. PMID- 9738671 TI - Neuronal complexity loss in interictal EEG recorded with foramen ovale electrodes predicts side of primary epileptogenic area in temporal lobe epilepsy: a replication study. AB - PURPOSE: To investigate whether a correct lateralization of the primary epileptogenic area by means of neuronal complexity loss analysis can be obtained from interictal EEG recordings using semi-invasive foramen ovale electrodes. In a previous study with recordings from intrahippocampal depth and subdural strip electrodes it was shown that the dynamics of the primary epileptogenic area can be characterized by an increased loss of neuronal complexity in patients with unilateral temporal lobe epilepsy (TLE). METHODS: Neuronal complexity loss analysis was applied. This analysis method is derived from the theory of nonlinear dynamics and provides a topological diagnosis even in cases where no actual seizure activity can be recorded. We examined interictal EEG recorded intracranially from multipolar foramen ovale electrodes in 19 patients with unilateral TLE undergoing presurgical evaluation. RESULTS: The primary epileptogenic area was correctly lateralized in 16 of the 19 investigated patients. The misclassification of the side of seizure onset in three patients might be attributed to the larger distance between the foramen ovale electrodes and the mesial temporal structures as compared to intrahippocampal depth electrodes. CONCLUSIONS: Our results confirm the previous findings and provide further evidence for the usefulness of nonlinear time-series analysis for the characterization of the spatiotemporal dynamics of the primary epileptogenic area in mesial temporal lobe epilepsy. PMID- 9738672 TI - Location of the epileptic zone and its physiopathological effects on memory related activity of the temporal lobe structures: a study with intracranial event related potentials. AB - PURPOSE: Neuropsychological research with epileptic patients has suggested that the location of seizure focus may be an important variable determining the nature and severity of memory impairments. According to this assumption, this study was designed to investigate the effects of the location of the epileptic zone on the memory-related activity recorded directly from different temporal lobe structures. METHODS: Intracranial event-related potentials (ERPs) were recorded during a continuous recognition memory task, which is known to elicit the modulation of N400 and P600 components (i.e., the ERP old/new effect). The patients were separated into three groups according to the location of their epileptic zone: unilateral temporal (UTE), temporal plus extratemporal (TEE), and bitemporal (BTE). Recordings were obtained from three temporal lobe structures: hippocampus, amygdala, and lateral temporal cortex. RESULTS: The results showed that in the hippocampus, the ERP old/new effect was abolished in the TEE group only. In the amygdala, although largely unaffected, the ERP modulation appeared to be more impaired in UTE patients. Contrasting with these data is the observation that the magnitude and reliability of the ERP old/new effect recorded at the neocortical level increases as the epileptic zone extends to the temporal lobes (i.e., BTE>TEE>UTE). CONCLUSIONS: It is concluded that the memory-related activity modulation of memory ERPs recorded from different temporal lobe structures is affected differently by the presence of an epileptic zone as a function of its location. The possible clinical implications of these findings in surgery planning are also discussed. PMID- 9738673 TI - Chromosome 20 ring: a chromosomal disorder associated with a particular electroclinical pattern. AB - PURPOSE: The chromosome 20 ring [r(20)] is a rare chromosomal disorder without clear phenotypical markers. We describe the electroclinical pattern in a group of patients with r(20). METHODS: We observed 3 patients (a boy, patient 1; his mother, patient 2; and an unrelated man, patient 3), performing prolonged video EEG and cytogenetic studies and fluorescent in situ hybridization (FISH) with chromosome-specific telomeric probes. RESULTS: All 3 patients had a very similar abnormal electroclinical pattern characterized by long bursts or trains of rhythmic theta waves, which were sharply contoured or had a notched appearance (with no detectable clinical correlate), and generalized spike waves (SW) associated with seizures of probable frontotemporal origin (SFT). In all 3 patients, the cytogenetic analysis of T lymphocytes showed mosaicism with a normal cell line and a second cell line with a chromosome 20, although the latter was little represented in patients 2 and 3. A few cells with a single chromosome 20 were also found. The same cytogenetic findings were confirmed in the lymphoblastoid cell line of patient 1 and in the fibroblasts of patient 3. FISH with chromosome-specific telomeric probes and TTAGGG sequences demonstrated the integrity of the ring chromosomes. CONCLUSIONS: The clinical picture of these patients appears to be related to the instability of the r(20)-generating cells monosomic for chromosome 20 and is thus haploinsufficient for a gene. In these patients, the electroclinical pattern of theta waves (probably unrelated to epilepsy) and the SW and SFT, even with mild mental retardation (MR) or no MR and without dysmorphic features, suggest that the r(20) syndrome may be present. PMID- 9738674 TI - Clobazam has equivalent efficacy to carbamazepine and phenytoin as monotherapy for childhood epilepsy. Canadian Study Group for Childhood Epilepsy. AB - PURPOSE: To compare the effectiveness of monotherapy clobazam (CLB) to carbamazepine (CBZ) and phenytoin (PHT) in children with epilepsy. METHODS: Children aged 2-16 years with newly diagnosed epilepsy or previous failure of one drug (for poor efficacy or side effects) were assigned to one of two study arms and then randomized--CLB versus CBZ or CLB versus PHT. Eligible children had partial epilepsies or only generalized tonic-clonic seizures. After a drug initiation protocol, monotherapy treatment mimicked the usual routines used by Canadian child neurologists. Blinding used a "double dummy" technique with blinded medication serum levels (6-point scale). Intention to treat analysis using survival curves assessed the primary endpoint--length of retention on the initial medication during the year after randomization. RESULTS: Fifteen centers entered 235 patients: 159 randomized to CLB versus CBZ and 76 to CLB versus PHT. Altogether, in all study arms, 119 received CLB, 78 CBZ, and 38 PHT. Overall, 56% continued to receive the original medication for 1 year with no difference between CLB and standard therapy (CBZ and PHT). Seizure control was equivalent for all three medications, as were side effects. PHT and CBZ induced more biologic side effects, such as rash, while CLB induced slightly more behavioral effects. Tolerance developed in 7.5% of patients receiving CLB, 4.2% with CBZ and 6.7% with PHT. CONCLUSIONS: CLB should be considered as "first line" monotherapy along with CBZ and PHT for all partial and selected generalized childhood epilepsies. PMID- 9738675 TI - Acute symptomatic seizure disorders in young children--a population study in southern Taiwan. AB - PURPOSE: To determine the incidence, etiology, and prognosis of acute symptomatic seizures in children by age 3 years. METHODS: In a population-based birth cohort study of all 15,209 neonatal survivors born in Tainan City between October 1989 and December 1991, parents or caretakers of 13,493 children aged 3 years were surveyed by telephone regarding any provoked convulsive disorder, particularly acute symptomatic seizure, in the children; medical records were reviewed. RESULTS: Sixty-three children (39 boys, 24 girls) had acute symptomatic seizures (incidence 0.46 in 100). The leading causes of acute symptomatic seizures were acute gastroenteritis, encephalitis/encephalopathy, and bacterial meningitis. Age specific incidence was highest in the group aged 1-12 months. Intracranial hemorrhage, bacterial meningitis, and metabolic disturbance were the major causes of acute symptomatic seizures in children aged 1-12 months. Acute gastroenteritis, encephalitis/encephalopathy, and bacterial meningitis accounted for 85% of the causes in children aged 13-24 months, and gastroenteritis and encephalitis/encephalopathy were the predominant causes in those aged 25-36 months. By age 5 years, subsequent unprovoked seizures developed in 14% of the survivors of acute symptomatic seizures. CONCLUSIONS: Many acute symptomatic seizures are preventable. The risk of subsequent unprovoked seizures is determined by underlying precipitating factors. Public education regarding the danger of shaken-baby syndrome and excessive water supplement, as well as and nationwide vaccination against bacterial meningitis in young children, is necessary. PMID- 9738676 TI - Evaluating health-related quality of life outcomes in clinical trials of antiepileptic drug therapy. AB - PURPOSE: To provide an overview of condition-specific health-related quality of life (HRQL) assessment in clinical trials of antiepileptic drug (AED) therapy in adults. We describe the key measurement issues in HRQL evaluation, identify the instruments that have been used in this population, summarize the psychometric characteristics of these instruments, propose areas of HRQL most likely to change with treatment, and offer recommendations for further research. METHODS: We conducted a comprehensive review of the literature using repeated searches of the MEDLINE database together with a review of reference lists from published papers. Psychometric information on the instruments was gathered from published literature. RESULTS: Three epilepsy-specific HRQL measures were identified: the Epilepsy-Surgery Inventory (ESI-55), the Liverpool Assessment Battery, and the Quality of Life in Epilepsy Inventory (QOLIE, the 89-, 31-, and 10-item versions). One new measure, the Epilepsy Foundation of America (EFA) Concerns Index was also found. The psychometric characteristics of these instruments are discussed in relationship to performance or expected performance in a clinical trial setting. A review of descriptive studies and trials to date suggests that subscales reflecting the psychological and social domains of HRQL may be most sensitive to treatment designed to increase seizure-free periods, reduce seizure severity, and minimize undesirable side effects. CONCLUSIONS: Although evaluation of HRQL outcomes in clinical trials of epilepsy is still in its infancy, several reliable and valid condition-specific measures are available for understanding the impact of disease and treatment on HRQL. Further research is needed to determine minimal clinically important change scores and to assess the psychometric stability of measures across cultures and mode of administration (self, interview, telephone). Studies of patient preferences for health outcomes in the form of utilities will provide needed data for evaluating the cost effectiveness of new treatments. PMID- 9738677 TI - Nonepileptic seizures after head injury. AB - PURPOSE: To examine the role of head injury as a risk factor in the development of nonepileptic seizures (NES). Specifically, we will determine the relative frequency of head injury among NES patients referred to our center and will describe several pertinent clinical features and personal characteristics. METHODS: Retrospective record review of patients referred to our center for evaluation of seizures over a 4-year period. All patients with NES were evaluated as in a previously described protocol, which included intensive video EEG monitoring, provocation by suggestion, and psychiatric interview. All NES patients with a history of head injury were extracted for this report. RESULTS: Of 102 patients with NES, nearly one-third (32%) had an antecedent head injury; 52% were male, mean age was 34 years, and 12% had coexisting epilepsy. Multiple psychiatric disorders were not uncommon (79%), and a history of abuse was found in 35%. All but four patients had documented financial gain from their injury. Follow-up at 1 year found poor long-term outcome with lasting disability; despite that, the majority (91%) of head injuries were minor. CONCLUSIONS: Our preliminary findings suggest that prior head injury is associated with the development of NES and may contribute to the pathogenesis of NES in vulnerable patients. Head injury and sexual or physical abuse appear to occur in comparable proportions in patients with NES. This suggests that head injury and abuse may be equally important risk factors in the development of NES. PMID- 9738678 TI - Brain blood flow alterations induced by therapeutic vagus nerve stimulation in partial epilepsy: I. Acute effects at high and low levels of stimulation. AB - PURPOSE: Left cervical vagus nerve stimulation (VNS) decreases complex partial seizures (CPS) by unknown mechanisms of action. We hypothesized that therapeutic VNS alters synaptic activities at vagal afferent terminations and in sites that receive polysynaptic projections from these medullary nuclei. METHODS: Ten patients with partial epilepsy underwent positron emission tomographic (PET) measurements of cerebral blood flow (BF) three times before and three times during VNS. Parameters for VNS were at high levels for 5 patients and at low levels for 5. Resting BF measurements were subtracted from measurements during VNS in each subject. Subtraction data were averaged in each of 2 groups of 5 patients. t Tests were applied to BF changes in brain regions that receive vagal afferents and projections (significant at p < 0.05, corrected for repeated measures). RESULTS: In both the low- and high-stimulation groups during VNS, brain BF was (a) increased in the rostral, dorsal-central medulla; (b) increased in the right postcentral gyrus, (c) increased bilaterally in the hypothalami, thalami, and insular cortices, and in cerebellar hemispheres inferiorly; and (d) decreased bilaterally in hippocampus, amygdala, and posterior cingulate gyri. The high-stimulation group had greater volumes of activation and deactivation sites. CONCLUSIONS: Our findings suggest that left cervical VNS acutely increases synaptic activity in structures directly innervated by central vagal structures and areas that process left-sided somatosensory information, but VNS also acutely alters synaptic activity in multiple limbic system structures bilaterally. These findings may reflect sites of therapeutic actions of VNS. PMID- 9738679 TI - Postictal nosewiping lateralizes and localizes to the ipsilateral temporal lobe. AB - PURPOSE: We observed many patients with temporal lobe epilepsy (TLE) wiping their nose postictally, usually with the hand ipsilateral to their seizure focus. We wished to determine if this had lateralizing or localizing significance. METHODS: We retrospectively studied 87 patients: 47 with unilateral TLE defined by successful surgical outcome [30 with medial TLE (MTLE) and 17 with neocortical TLE (neoTLE)]; and 40 with extratemporal epilepsy (ExTLE). Videotapes of 319 complex partial seizures (CPS) without generalization were reviewed by 1 neurologist, blinded to patient identity, who recorded each episode of nosewiping and the hand with which it was performed. RESULTS: With regard to localizing potential, postictal nosewiping (PINW) was significantly more common in patients with unilateral TLE than in those with ExTLE. In the TLE group, PINW within 60 s of electrographic seizure offset occurred in 60% of patients (28 of 47) and 43% of seizures (74 of 171). In the ExTLE group, PINW was noted in 33% of patients (13 of 40; p < 0.05 as compared with TLE) and 15% of seizures (22 of 148; p < 0.001). Similar results were obtained with PINW within 30 s of seizure offset. Although PINW was more frequent in MTLE than in neoTLE (67% of patients vs. 47%), this finding did not reach significance. With regard to lateralizing potential, in the TLE group, unilateral PINW (performed with a single hand only) within 60 s of seizure offset was observed in 53% of patients (25 of 47) and was performed with the hand ipsilateral to the seizure focus in 92% (23 of 25). Thirteen patients (9 with TLE) wiped their nose more than once with the same hand in a single seizure within 60 s of offset in 18 seizures; this was done with the hand ipsilateral to the seizure focus in all 18 instances (predictive value = 100%). CONCLUSIONS: PINW is more common in unilateral TLE, particularly MTLE, than in ExTLE. PINW performed exclusively with one hand occurs in approximately 50% of patients with TLE and is highly predictive (92%) of seizure onset ipsilateral to the hand used, especially when it occurs repetitively. We hypothesize that ictal activation of the central autonomic nervous system, particularly the amygdala, results in ictal nasal secretions and causes nosewiping as the patient regains awareness postictally. The ipsilateral hand is used due to contralateral neglect or weakness. PMID- 9738680 TI - Aspiration: a potential complication to vagus nerve stimulation. AB - PURPOSE: Vagus nerve stimulation (VNS) is reported to reduce the frequency of seizures in children and adults without causing serious side effects. However, clinical observation of swallowing difficulties in 2 children treated with VNS made further investigation necessary. METHODS: Seven patients aged 4-18 years and treated with VNS for 6-14 months were investigated with videoradiography during barium swallow. The children performed 5-30 barium swallow investigations with the VNS device turned off, running as programmed, or set at continuous stimulations. The degree of aspiration was scored from 0 to 3. RESULTS: In 5 of 7 children, of whom reported transient swallowing difficulties, no change in the degree of aspiration was noted. The 2 children with swallowing difficulties, however. showed increased aspiration score when the stimulator was set at continuous stimulations. In 1 the score also appeared to increase with the VNS running as programmed (p > 0.05). Both children had severe mental and motor disabilities. CONCLUSIONS: Before and during VNS treatment patients should be evaluated with regard to swallowing problems. There needs to be an easy way to turn the device on and off to avoid aspirations, a hazardous and potentially life threatening complication of VNS. PMID- 9738681 TI - Single photon emission computed tomography (SPECT) in a patient with bilateral temporal seizures: correlation between ictal EEG and postictal/ictal SPECT. AB - We report a patient with bilateral independent temporal lobe seizures in whom two [99mTc]HMPAO single photon emission computed tomograph (SPECT) scans were performed during two different seizures. In the first periictal SPECT, [99mTc]HMPAO was injected in the interval between two closely spaced seizures (one localized in the left temporal lobe and the other in the right temporal lobe). SPECT images showed hypoperfusion in the left lateral temporal lobe, hyperperfusion of the left mesial temporal region, and pronounced hyperperfusion in the right anterior temporal lobe. These results suggest both a postictal left temporal SPECT pattern and an ictal right temporal pattern. In the second periictal SPECT, [99mTc]HMPAO was injected immediately after a right temporal lobe seizure and showed right lateral temporal lobe hypoperfusion and right mesial hyperperfusion, suggesting a postictal right temporal SPECT pattern. Interpretation of the periictal SPECT should take into account EEG changes at the time or in the minutes immediately after injection of [99mTc]HMPAO. PMID- 9738682 TI - Semiological seizure classification. AB - We propose an epileptic seizure classification based exclusively on ictal semiology. In this semiological seizure classification (SSC), seizures are classified as follows: a. Auras are ictal manifestations having sensory, psychosensory, and experiential symptoms. b. Autonomic seizures are seizures in which the main ictal manifestations are objectively documented autonomic alterations. c. "Dialeptic" seizures have as their main ictal manifestations an alteration of consciousness that is independent of ictal EEG manifestations. The new term "dialeptic" seizure has been coined to differentiate this concept from absence seizures (dialeptic seizures with a generalized ictal EEG) and complex partial seizures (dialeptic seizures with a focal ictal EEG). d. Motor seizures are characterized mainly by motor symptoms and are subclassified as simple or complex. Simple motor seizures are characterized by simple, unnatural movements that can be elicited by electrical stimulation of the primary and supplementary motor area (myoclonic, tonic, clonic and tonic-clonic, versive). Complex motor seizures are characterized by complex motor movements that resemble natural movements but that occur in an inappropriate setting ("automatisms"). e. Special seizures include seizures characterized by "negative" features (atonic, astatic, hypomotor, akinetic, and aphasic seizures). The SSC identifies in detail the somatotopic distribution of the ictal semiology as well as the seizure evolution. The advantages of a pure SSC, as opposed to the current classification of the International League Against Epilepsy (ILAE), which is actually a classification of electroclinical syndromes, are discussed. PMID- 9738683 TI - Classifications of the International League Against Epilepsy: time for reappraisal. AB - Recent advances in neurodiagnostic technology, new insights into fundamental neuronal mechanisms of epilepsy, and rapid developments in molecular genetics have greatly improved our understanding of epileptic seizures and epileptic disorders since 1981, when the current International Classification of Epileptic Seizures was adopted, and since 1989, when the International Classification of Epilepsies, Epileptic Syndromes, and Related Disorders was adopted. Although both have been universally accepted and have proven to be of considerable clinical value, it has become clear that more is needed for specific applications of growing importance, such as presurgical evaluation, clinical pharmacology trials, and epidemiological studies. Therefore, an International League Against Epilepsy (ILAE) task force has evaluated the need for revising our current classification and terminology and has begun developing four specific documents. The first is a descriptive terminology for ictal phenomena, incorporating some of the concepts included in the preceding paper by Dr. Hans Luders; however, this will be a glossary, not a classification. The second is a classification of epileptic seizures based on known or presumed pathophysiological and anatomic substrates, to replace the current classification, which is based entirely on phenomenology. The third is a classification of epileptic syndromes and epileptic diseases, adapted from the current Classification of Epilepsies, Epileptic Syndromes, and Related Disorders, which will take into account the fact that disorders with unique etiologies, such as a single gene, should be considered diseases, and which will be reorganized for maximum practical application to differential diagnosis. The fourth is a new classification of functional disability due to seizures or epilepsy, based on a new impairment classification for neurological disorders being developed by the World Health Organization. Working groups are currently in the process of developing each of these documents, and their chairs would benefit from any comments or suggestions from those who ultimately will be using these systems of classification and terminology. PMID- 9738684 TI - Vagus nerve stimulation (VNS) may be useful in treating patients with symptomatic generalized epilepsy. PMID- 9738685 TI - The label "intellectually challenged" is politically correct, but what does it mean factually and medically? PMID- 9738686 TI - A call for caution in the interpretation of the observed smaller relative importance of risk factors in the elderly. PMID- 9738687 TI - The effect of age on risk factors for ischemic heart disease: the Manitoba Follow Up Study, 1948-1993. AB - PURPOSE: The purpose of this paper is to determine the age-specific relationships between risk factors at age 40 through 75 years and ischemic heart disease (IHD), and to determine the effects of aging on these relationships in a cohort of 3983 Canadian males. METHODS: The Manitoba Follow-Up Study is the prospective investigation of cardiovascular disease as it develops in a cohort of 3983 young men. Over a period of 45 years, from 1948 to 1993, 1094 study members (27%) developed clinical evidence of IHD. Blood pressure, body weight, smoking, and presence of diabetes mellitus have been recorded at regular intervals throughout the follow-up period. Using measurements from examinations every 5 years between ages 40 and 75 years, age-specific Cox proportional hazard models were fit to relate these risk factors to IHD. RESULTS: The adjusted relative risk of IHD for systolic blood pressure, diastolic blood pressure and smoking were found to significantly (p < 0.001) decline with advancing age. The adjusted relative risk for body mass index and presence of diabetes mellitus for ischemic heart disease did not vary with age (p > 0.05). After age 65 years, these risk factors were of little value for the prediction of IHD. CONCLUSIONS: The relative risk and statistical significance of blood pressure and smoking, as risk factors for IHD, decline with age. PMID- 9738688 TI - Prevalence and correlates of overweight among elementary schoolchildren in multiethnic, low income, inner-city neighbourhoods in Montreal, Canada. AB - PURPOSE: Increased understanding of the early determinants of obesity is essential because of the increasing prevalence of obesity in many industrialized countries. METHOD: As part of the evaluation of a school-based heart health promotion intervention, we measured height, weight, and triceps skinfold thickness at baseline in 2108 students aged 9-12 years (80.5% of eligible students) in 24 inner-city elementary schools located in multiethnic, low income neighbourhoods in Montreal, Canada. Data on student's socio-demographic and lifestyle characteristics were collected in classroom-administered questionnaires, and parents completed an at-home self-administered questionnaire. RESULTS: Overall, 35.2% of boys and 33.0% of girls were overweight (> or = 85th age and gender-specific percentiles from NHANES 11, for body mass index and triceps skinfold thickness); 15.1% of boys and 13.3% of girls were obese (> or = 95th age and gender-specific percentiles for body mass index and triceps skinfold thickness). Younger age, having lived all one's life in Canada, and being of European or Central American/Caribbean family origin were independent correlates of obesity in boys. Younger age, ever smoked, mother obese and father obese were independent correlates of obesity in girls. Girls of Asian family origin were protected. CONCLUSIONS: The very high prevalence of overweight students in this low income, multiethnic population suggests an important need for preventive intervention. PMID- 9738689 TI - Relationship between changes in dietary sucrose and high density lipoprotein cholesterol: the CARDIA study. Coronary Artery Risk Development in Young Adults. AB - PURPOSE: Cross-sectional data from several observational studies have suggested that dietary sucrose may be inversely associated with high density lipoprotein cholesterol (HDL-C). This study examined associations between energy from dietary sucrose and HDL-C at baseline, year 7 and longitudinally (year 7 minus baseline) in a cohort of young black and white men and women from the Coronary Artery Risk Development in Young Adults (CARDIA) study. METHODS: The sample included 4734 black men, black women, white men and white women, ages 18-30 years, in 1985-86 (baseline); 3513 at year 7; and 3335 for longitudinal analyses. Multivariate analyses was used with adjustment for age, BMI, cigarettes smoked per day, physical activity score, and alcohol intake. RESULTS: Multivariate analyses indicated that energy intake from sucrose was inversely associated with HDL-C for each race-gender group at baseline, year 7, and longitudinally from baseline to year 7. This association was significant at baseline for black men, and white men and women (p < 0.01); at year 7 for white men and black women (p < 0.01), and longitudinally for white men, white women, and black women (p < 0.05). CONCLUSIONS: The consistent inverse associations between energy from dietary sucrose and HDL-C observed in both cross-sectional and longitudinal analyses, and in different race and gender groups in CARDIA suggest that lowering dietary sucrose intake may be beneficial for those who may have low HDL-C. PMID- 9738690 TI - Mammography use by elderly women: a methodological comparison of two national data sources. AB - PURPOSE: Estimates of mammography utilization vary considerably, depending on the data source. Among women aged 65 years and older, recent estimates of annual mammography derived from the 1992 National Health Interview Survey (NHIS) were 50% higher than estimates from Medicare claims. We investigated possible reasons for the different estimates. METHODS: We identified differences in the populations covered by the two data sources and made appropriate adjustments. Differences due to age were addressed by age restriction and age adjustment. Women in health maintenance organizations were eliminated from the NHIS sample so it more closely resembled the Medicare database, and estimates of mammography utilization by noninstitutionalized women were derived for Medicare to increase comparability with NHIS. By using the Medicare Current Beneficiary Survey to obtain individual-level comparisons between self-report and claims, we explored potential biases in self-reported data and missing claims. RESULTS: Differences between the sample populations accounted for more than one-fourth of the rate difference. About half of the difference could be attributed to erroneous self reports, biases in self-reported dates (forward and reverse telescoping) and missing Medicare claims. CONCLUSIONS: Most of the discrepancy between the two data sources can be plausibly explained. However, caution must be used in using either data source alone, or both together, to represent the "true" mammography rate. PMID- 9738691 TI - The effects of dehydroepiandrosterone (DHEA) on the thymus, spleen, and adrenals of prepubertal and adult female rats. AB - Administration of dehydroepiandrosterone (DHEA) to female rats produces a condition of reproductive failure and ovarian cysts similar to that seen in women having polycystic ovarian disease. On the other hand, DHEA may have beneficial effects on the immune system. We sought to determine the effect of DHEA, when administered in pharmacological amounts, on the thymus and spleen of prepubertal (25 day old) and young adult (60 day old) female rats. Since the adrenal, by means of its production of corticosteroids, also is known to modulate the immune system, we also evaluated the effects of DHEA administration on this gland. The daily SC administration of DHEA (6 mg/100g BW) to young adult female rats led to progressive and striking reductions in thymic weights (greater than 85% suppression after 20 days compared to vehicle treated animals). There were no effects of DHEA on body weights or the weights of the spleen. DHEA treatment also led to significantly reduced weights of the adrenals , which was sustained at about 15-20% below normal over 5-20 days treatment. Ovariectomy of the rats 5 days before initiation of DHEA or vehicle treatment gave rise to significant increases in thymic and spleenic weights in control animals and strikingly blunted the inhibitory effects of DHEA treatment for 10 days on the thymus; DHEA had no effect on the ovariectomy-induced rise in the weight of the spleen. Ovariectomy also had no effect on the inhibitory effects of DHEA on adrenal weight. Similar, albeit quantitatively less striking, responses were noted to occur after DHEA treatment in immature female rats. These data indicate that DHEA in doses sufficient to interfere with ovarian cyclicity also has potentially adverse effects on the adrenal and thymus. The ovary appears to play an independent role in maintenance of the size of the thymus and spleen and also may mediate some of the effects of DHEA on the thymus but not those on the adrenals. PMID- 9738693 TI - Effects of rhG-CSF on neutrophil functions and survival in sepsis induced diabetic rats. AB - Diabetic patients are more prone to infection and evidence for an immunologic defect superimposed upon the metabolic abnormalities of diabetes is convincing. Neutrophils play a critical role in the host defense mechanism against various bacterial infections, and it is suggested that impaired neutrophil functions cause susceptibility to infections in diabetic patients. To explore the possibility that Granulocyte colony-stimulating factor (G-CSF) may be useful to prevent the morbidity and mortality caused by infections in diabetics. We studied the effect of G-CSF against septicemia in diabetic rats. Forty eight rats were divided into seven equal groups. The IInd, IVth-VIth groups were made diabetic by single intraperitoneal injection of streptozotocin. Fourth, VIth and VIIth groups were made septicemic by cecal ligation and perforation at the end of the second week of streptozotocin injection. G-CSF was subcutaneously injected into IIIrd, Vth and VIth groups. White blood cell count, neutrophil counts and function were determined. Rats in all groups were also observed for seven days for survival. White blood cells, neutrophil and lymphocyte counts and the neutrophil phagocytosis index decreased but neutrophil adherence rate was not different in diabetic group II (p<0.05). All these variables were significantly diminished in diabetes and sepsis-induced group IV (p<0.05). G-CSF injections improved all variables except neutrophil adherence. Cumulative survival ratio was better in G CSF-injected group VI than in ceftriaxon-administrated group VII (p<0.05). In conclusion, G-CSF increased neutrophil counts, developed neutrophil functions and improved survival. These results suggest that G-CSF may be useful as a drug to prevent bacterial infection in diabetic patients. PMID- 9738692 TI - Dissociation of vascular and adrenal responsiveness to angiotensin II following calcium channel blockade. AB - Calcium channel blockers as a class reduce both vascular resistance and systemic blood pressure. However, it is not known if different classes of calcium channel blockers have similar effects on the renin-angiotensin-aldosterone system. We investigated vascular and adrenal responses to endogenous and exogenous angiotensin II in normotensive subjects before and after receiving isradipine or diltiazem. Subjects achieved low salt balance before study and underwent an angiotensin II infusion and upright posture study. After re-achieving salt balance the subjects received either isradipine (n=10), or diltiazem (n=7) for three days before repeating the study. Both agents lowered blood pressure and significantly shifted the dose response relationship of angiotensin II and mean blood pressure (P<0.01). In contrast, aldosterone secretion in response to upright posture and angiotensin II infusion was significantly reduced (P<0.01) by isradipine but not by diltiazem. The in vitro effects of both agents on aldosterone secretion from bovine adrenal glomerulosa cells paralleled the in vivo observations showing that angiotensin II-stimulated aldosterone secretion was markedly blunted by isradipine and minimally by diltiazem. These results suggest a tissue specificity for the effects of different classes of calcium channel blockers. Dihydropyridine calcium channel blockers may exert their antihypertensive effect by blocking both vascular and adrenal responses to angiotensin II. PMID- 9738694 TI - Estrogen, androgen and antiestrogen responses in the accessory organs of male rats during different phases of life. AB - Recent studies have indicated that estrogen has a stimulatory influence on the male reproductive tract. Evidence includes the presence of measurable levels of estrogen in the circulation, retention of exogenous estrogen, and presence of estrogen receptors in the male accessory sex organs during prepubertal life. In the present study, estrogen antagonists (CDRI-85/287 and centchroman) have been used to examine this concept by antagonising estrogen action at critical stages in the life in rat. Centchroman or 85/287 administration to 14 day old rats for 7 days did not alter gonadal or accessory organ weight. In contrast, in 21 day old castrated rats, treatment with either compound from day 28-35 of life stimulated an increase in all organ weights. When administered to normal rats during the critical phase of transition, i.e. days 30-60 of life, both testis and accessory organs showed an increase in weight. In contrast castrated rats treated with estrogen alone or in combination with 85/287 from days 37-45 of life and sacrificed on day 46 did not show any change, but 85/287 per se markedly reduced the weight of accessory organs. In adult castrated rats, the potency of DHT as a promoter of growth was potentiated by estradiol. Compound 85/287 negated the estradiol-induced increase. Glycerylphosphorylcholine (GPC) and sialic acid levels showed about 100% increase, with both high and low doses of 85/287 (treated from 30-60 days of life), However, centchroman (CDRI-67/20) was less potent in this regard. The effect of estrogen antagonists in relation to epididymal physiology during different phases of life in the rat is discussed. PMID- 9738696 TI - Antineutrophil cytoplasmic antibodies in autoimmune thyroid disorders. AB - Different antibodies against both organ- and non-organ-specific autoantigens have been found in patients with autoimmune thyroid diseases. The aim of our study was to evaluate the presence of antineutrophil cytoplasmic antibodies (ANCA) in sera of patients affected by Graves' disease (GD) and Hashimoto's thyroiditis (HT). These antibodies were investigated by indirect immunofluorescence; the reactivity against myeloperoxidase and lactoferrin was assessed by ELISA. ANCA were detected by immunofluorescence in 28.5% of patients with GD and 9% of patients with HT. Anti-lactoferrin antibodies were found in 3 of 21 (14.2%) patients affected by GD and in 2 of 11 (18.1%) cases of HT. Anti-myeloperoxidase antibodies were detected only in one (4.7%) patient with GD. PMID- 9738695 TI - Effect of CDRI-85/287 on uterine estradiol and progesterone receptor levels/morphometric measurements during preimplantation period in rat. AB - Studies with 85/287, a potent nonsteroidal antiestrogen/antiimplantation agent were taken up. In this paper we report alterations in uterine morphometric measurements and estrogen/progesterone receptor levels under the influence of this compound. Results showed 32% decline in stromal absolute volume density on day 5 post-coitum (p.c.) only, whereas eosinophilic leucocyte number decreased both on days 3 and 5 p.c. (41%) in treated rat uterus. Epithelial mitotic activity showed complete cessation both in control and treated rats on day 5 p.c. Under the influence of the compound both cytosolic and nuclear estrogen receptor (ERc and ERn) levels decreased on day 3 p.c., but on day 5 p.c. ERn increased significantly. A significant increase was however noticed in progesterone nuclear receptors (PRn) on day 5 p.c. On the whole our studies showed overall significant changes in uterine morphometric measurement/estrogen and progesterone receptor levels during the preimplantation period in rat under the influence of compound 85/287, causing asynchrony of events and thus failure of implantation. PMID- 9738697 TI - In vitro production of progesterone and estradiol by rat granulosa cells regulated by cabergoline and prolactin. AB - Cabergoline (CG), a dopamine agonist, and prolactin (PRL) added in vitro to isolated granulosa cells from immature rats cultured in serum-free medium inhibited FSH-stimulated production of progesterone. The lowest effective concentrations of CG and PRL were 10(-8)M and 1 ng/mL, respectively. In combination, the inhibitory activities were additive. CG at 10(-8) M also suppressed estradiol production by isolated rat granulosa cells; whereas PRL did not. In conclusion, CG inhibits FSH-stimulated progesterone and estradiol production by rat granulosa cells. PMID- 9738699 TI - Association of insulin resistance to electrocardiographic changes in non obese Asian Indian subjects with hypertension. AB - To investigate the relationship between insulin resistance and electrocardiographic changes in hypertension in the absence of confounding influences, plasma glucose and insulin responses to oral glucose were studied in 26 normotensive and 38 hypertensive subjects. Resting ECG was taken and classified into normal or abnormal using the Minnesota code. Among the 38 subjects, 16 had ECG abnormalities. All the hypertensive subjects had normal glucose tolerance. Serum insulin response of hypertensive subjects with ECG changes was 43% higher than those of hypertensive subjects without ECG changes and of normotensive subjects. The ratio AUC glucose/AUC insulin, a measure of insulin sensitivity was significantly reduced in subjects with abnormal ECG. Serum LDL cholesterol was significantly elevated and was the highest in hypertensive subjects with abnormal ECG. The ratio, Total Cholesterol/HDL Cholesterol was elevated to 5.81+/-0.47. I(125)-insulin binding to erythrocytes from 6 normotensive subjects, and 16 hypertensive subjects (8 with and 8 without ECG abnormalities) indicated 50% reduction in insulin receptor number in both the groups of hypertensive subjects compared to normotensive subjects. Multiple logistic regression analysis using mean blood pressure, serum total cholesterol, LDL cholesterol/HDL cholesterol, sex, insulin level at 60 min in OGTT, treatment, serum triglyceride, presence of family history of diabetes, CHD, hypertension and tobacco as independent variables causing ECG changes, revealed correct classification in 84% of cases. Among the variables, insulin level in OGTT contributed the most to ECG abnormalities. The data suggest that in the non obese, non diabetic Asian Indian hypertensive subjects, the presence of electrocardiographic abnormalities might be partly related to hyperinsulinemia or insulin resistance in them. PMID- 9738698 TI - Low T3 syndrome with asynchronous changes of TT3 and rT3 values in laparoscopic cholecystectomy. AB - Non-thyroidal illnesses, such as surgical stress, are associated with abnormal metabolism of thyroid hormones. However, the potential impact of variable surgical procedures remain to be elucidated. In order to evaluate the effect of mild surgical stress upon thyroid function, TT4, TT3, rT3 and TSH were measured in twenty-two patients undergoing laparoscopic cholecystectomy before (Stage 1), during (Stages 2-5), at the recovery room (Stage 6) and 24h postoperatively (Stage 7). The values of TSH remained within the normal limits with transient changes during the study period. Similarly, TT4 values displayed normal variations within the normal range without reaching a statistically significant difference during the study period. A decrease of TT3 values was detected early at stage 2 during induction of anaesthesia. TT3 remained at low levels during the perioperative period, and a further decrease was observed 24 h postoperatively. The above profile of thyroid hormone metabolism, reflects a low-T3 syndrome in patients undergoing laparoscopic cholecystectomy. Interestingly, there was a tendency for rT3 to increase and it reach its highest value 24h postoperatively with the difference being statistically significant (p<0.05). The asynchronous distribution of rT3 and TT3 might be attributed to multifactorial influences. PMID- 9738700 TI - Acute effects of recombinant murine leptin on rat pituitary-adrenocortical function. AB - Leptin, the product of the ob gene, is an adipose-tissue secreted hormone, which regulates satiety, metabolic rate and thermogenesis. Many lines of evidence suggest the existence of mutual relationships between leptin and adrenal-cortex secretion, but in vivo studies gave rather conflicting results. Therefore, we have investigated the acute effect of the systemic bolus administration of leptin (5 or 10 nmol/kg) on the function of rat pituitary-adrenocortical function. The blood concentrations of ACTH, aldosterone and corticosterone were measured by specific RIA 60 or 120 min after the leptin injection. Leptin did not affect the blood concentrations of ACTH and aldosterone at 60 min, but at 120 min the lower dose of the peptide increased them. In contrast, the blood level of corticosterone was markedly enhanced by both doses of leptin at 60 and 120 min. Collectively, these findings indicate that leptin exerts a moderate acute activation of the pituitary-adrenocortical function in rats. They also suggest that the adrenocortical secretagogue action of leptin is not only mediated by the enhanced pituitary ACTH release, but is also consequent to a direct stimulatory effect of the peptide on adrenocortical cells. PMID- 9738701 TI - Serum leptin and IGF-I levels in cystic fibrosis. AB - The role of leptin in states of negative energy balance such as cystic fibrosis (CF) has not been explored. We hypothesized that leptin levels in patients with CF would be low due to correlation with body weight. Despite the importance of IGF-I in normal growth and anabolism, there are few data on IGF-I in CF. We studied 27 CF patients (25+/-5 yrs, 57+/-9 kg, 10M/17F) and 12 control subjects (25+/-4 yrs, 57+/-9 kg, 6M/6F). Each subject underwent analysis of lean body mass (LBM) and percent body fat by dual-energy x-ray absorptiometry (DEXA). Serum leptin and IGF-I levels were measured by radioimmunoassay. Serum leptin levels were similar both in CF and in controls (CF=5.3+/-4.1 ng/ml, C=4.4+/-3.6ng/ml; p=0.3), and there was no difference in percent body fat between the two groups (CF=26+/-13%, C=21+/-7%; p=0.3). Leptin levels were significantly lower in CF males than females corresponding to lower fat levels in males in both CF and controls. Leptin levels were positively correlated with percent body fat both in CF and controls (CF: r=0.8; p=0.01, CONTROL: r=0.8; p =0.2). Serum IGF-I levels were significantly lower in CF patients than in controls (CF=1.13+/-0.41 ng/ml, C=6.72+/-3.62 ng/ml; p=<0.01). We conclude that the physiological regulation of leptin is maintained in relation to body fat even in chronic illness and that the negative energy balance in CF is not caused by high leptin levels. PMID- 9738702 TI - Urinary collagen crosslinks reflect further bone loss of femoral neck in osteoporotic patients undergoing vitamin D therapy. AB - Bone mineral density (BMD) of the lumbar (L2-LA) spine and femoral neck was measured annually for 2 years (3 times beginning at the beginning of year 1 and after each subsequent year) in 39 female patients with osteoporosis undergoing 0.5 or 1.0 microg daily doses of vitamin D therapy. At the time of the first BMD measurement, biochemical markers including serum alkaline phosphatase (ALP), urinary pyridinoline (Pyr), deoxypyridinoline (Dpyr) and hydroxyproline (Hyp) were also measured. Urinary Pyr and Dpyr correlated with the percent changes of femoral neck BMD measurements taken the year following the sampling (Pyr: r= 0.622, p<0.001, Dpyr: r=-0.385, p<0.05). Only urinary Pyr correlated with the percent changes of femoral neck BMD measurements taken the following 2 years (r= 0.532, p<0.05). Neither serum ALP nor urinary Hyp correlated with the percent changes of spine or femoral neck BMD measurements taken the year or 2 years after the sera and urine sampling. In summary, we believe urinary Pyr and Dpyr can reflect subsequent bone loss of the femoral neck BMD having been measured only once during the course of a year. PMID- 9738703 TI - Effect of chronic thyroxine treatment on pregnancy in rats: effects on oestrogen, progesterone, prolactin and GH receptors in uterus, liver and mammary gland. AB - We have previously shown that experimental hyperthyroidism produces premature and difficult delivery and absence of lactation in spite of apparently adequate luteolysis and lactogenesis. To study the possible causes of these alterations we measured the effect of treatment with T4 (0.25 or 1 mg kg(-1), s.c., daily, started 10-15 days before mating, HT0.25 and HT1) on serum hormones and their receptor (R) concentrations in reproductive tissues on day 20 of pregnancy (1800 hours), comparing them with controls on the same day (C20), or on day 21 of pregnancy (1800 hours) (C21). Serum prolactin (PRL) and corticosterone (B) concentrations increased in the HT groups, progesterone (Pg) and GH decreased and estradiol (E2) did not change, compared with C20 group. C21 rats had increased serum PRL and decreased Pg and GH. In HT rats mammary DNA and protein tissue content was doubled. Receptor concentrations were expressed per mg DNA. Mammary PRL-R were increased in HT1 rats, while E-R and Pg-R were significantly lower in both HT groups. HT0.25 and HT1 rats had increased uterine E-R and Pg-R and decreased liver PRL-R and GH-R as well as their mRNAs. Liver E-R, PRL-R and GH-R were decreased in C21 rats, while uterine Pg-R were increased. Thus, some of the observed changes (serum Pg and GH, mammary and uterine Pg-R, and liver GH-R and PRL-R decreases and serum PRL increase) may be due at least partially to the advancement in luteolysis and delivery, being similar to the changes observed between days 20 and 21. The changes in serum B, mammary PRL-R, and mammary and uterine E-R may be caused solely by the T4 treatments and may play a role in the alterations previously observed. PMID- 9738705 TI - Treatment of children with immune thrombocytopenic purpura: are we closer to resolving the dilemma? PMID- 9738704 TI - Serial analysis of circulating T gamma/delta lymphocyte subpopulations in Graves' disease. AB - In the present paper the distribution of peripheral blood T gamma/delta lymphocytes in Graves' disease patients is analyzed in order to correlate them with disease activity and with prognosis. Eighteen patients with Graves' disease, 24 patients with Hashimoto's thyroiditis and 32 sex- and age-matched healthy control subjects were studied. Peripheral blood CD3+ alpha/beta and gamma/delta T lymphocytes as well as CD4+ and CD8+ T cells, and CD19 (B) lymphocytes were analyzed by cytofluorometry. At diagnosis, patients who required a radical treatment for thyrotoxicosis control showed a significant decrease of T gamma/delta lymphocytes and an increase of B cells when compared with those who maintained the euthyroid state after methimazole. No correlation was detected between the percentages of these subpopulations and serum free thyroxine. This decreased proportion did not normalize after methimazole or radical treatments. These results suggest that the cytotoxic T gamma/delta compartment of the immune system is altered in patients with Graves' disease. PMID- 9738706 TI - Timeliness of codifying nutrition ABCDE's for BPD. PMID- 9738707 TI - Biomarkers of fetal exposure to alcohol: identification of at-risk pregnancies. PMID- 9738708 TI - Universal screening of infants for hearing loss: further justification. PMID- 9738709 TI - Learning disabilities just don't add up. PMID- 9738710 TI - Autosomal dominant granulomatous disease of childhood: the naming of things. PMID- 9738711 TI - Inroads in transcatheter therapy for congenital heart disease. PMID- 9738712 TI - Major hemorrhage in children with idiopathic thrombocytopenic purpura: immediate response to therapy and long-term outcome. AB - OBJECTIVES: We retrospectively characterized children with idiopathic thrombocytopenic purpura (ITP) who had major hemorrhage to determine response to therapy and long-term outcome. STUDY DESIGN: We reviewed the medical records of 332 children with ITP diagnosed at our center during the last 10 years for occurrence of major hemorrhage, defined as (1) intracranial hemorrhage, (2) epistaxis requiring cautery or nasal packing, (3) gross hematuria, or (4) other bleeding causing a decline in hemoglobin concentration. RESULTS: Of 332 patients with ITP, 58 (17%) had 68 episodes of major hemorrhage; 56 of these episodes were treated with corticosteroids, intravenous immunoglobulin, or both. The platelet count rose to > or =20,000/mm3 within 24 hours after presentation after only 18% of evaluated events, and 28% of patients with major hemorrhage still had a platelet count <20,000/mm3 after 7 days. Twenty-seven of 49 patients available for evaluation had resolution of ITP within 6 months, 21 had chronic ITP, and 1 died of sepsis. CONCLUSIONS: We observed that 17% of children with ITP had major hemorrhage. Only a minority of these patients had an immediate rise in platelet count after receiving intravenous immunoglobulin, corticosteroid treatment, or both. Prospective studies of childhood ITP focusing on short-term outcome variables in addition to platelet count should be performed to better define optimal treatment for each affected child. PMID- 9738713 TI - Growth and body composition in infants with bronchopulmonary dysplasia up to 3 months corrected age: a randomized trial of a high-energy nutrient-enriched formula fed after hospital discharge. AB - OBJECTIVES: (1) To determine whether nutrient malabsorption or inadequate nutrient intake were involved in the cause of growth delay in patients with bronchopulmonary dysplasia, and (2) to determine whether a nutrient-enriched formula given to infants with bronchopulmonary dysplasia to 3 months corrected age improves the rate of growth with greater lean and bone mass accretion when compared with infants fed an isoenergetic standard infant formula. STUDY DESIGN: A blinded, nutrition intervention trial of 60 preterm infants with bronchopulmonary dysplasia (birth weight, 866 +/- 169 g, gestational age, 26 +/- 1.5 weeks) randomized to either nutrient-enriched formula or standard formula. Growth, body composition, and nutrient retention were compared between groups by Student's t tests and analysis of covariance. RESULTS: Infants fed the enriched formula had significantly greater nitrogen and mineral retention at 38 weeks' postmenstrual age, and only the infants fed enriched formula had zinc retention similar to the intrauterine accretion. At 3 months corrected age infants fed enriched formula attained greater length (P < .05), greater radial bone mineral content (P < .01), and greater lean mass (P < .01). The male infants in the enriched formula group had greater whole body bone mineral content than did male infants in the standard formula group (P = .02). CONCLUSIONS: Greater linear growth and lean and bone mass in the enriched formula group suggests that infants with bronchopulmonary dysplasia attain faster "catch-up" growth when fed higher intakes of protein, calcium, phosphorus, and zinc than provided in standard proprietary formulas. PMID- 9738714 TI - The prenatal detection of significant alcohol exposure with maternal blood markers. AB - OBJECTIVE: To examine the efficacy of a combination of 4 blood markers of alcohol use in detecting alcohol-abusing pregnant women. STUDY DESIGN: Two new markers of alcohol use, whole blood-associated acetaldehyde and carbohydrate-deficient transferrin, and 2 traditional markers of alcohol use, gamma-glutamyl transpeptidase and mean red blood cell volume, were measured in the blood of pregnant women. Each woman was interviewed about alcohol and drug use, medical and obstetric histories, and nutrition. Each infant was examined by a clinician who was blinded to exposure status. RESULTS: All of the women who reported drinking an average of 1 or more ounces of absolute alcohol per day had at least 1 positive blood marker. The infants of mothers with 2 or more positive markers had significantly smaller birth weights, lengths, and head circumferences than the infants with negative maternal screens. The presence of 2 or more positive markers was more predictive of infant outcome than any self-reporting measure. CONCLUSIONS: These markers, which detect more at-risk pregnant women than self reporting methods, could lead to better efforts at detection and prevention of alcohol-induced fetal damage. PMID- 9738715 TI - The Rhode Island Hearing Assessment Program: experience with statewide hearing screening (1993-1996) AB - OBJECTIVE: The objective of this study was to evaluate key outcomes of a universal hearing screen/rescreen program for all births with transient evoked otoacoustic emissions in all 8 maternity hospitals in the state of Rhode Island over a 4-year period. STUDY DESIGN: This was a retrospective analysis of the hearing screen/rescreen refer data collected prospectively for 53,121 survivors born in Rhode Island between January 1, 1993, and December 31, 1996. Primary outcomes included the first-stage refer rates, rescreen compliance, diagnostic referral rates, identification rates, and the age of amplification. RESULTS: During this 4-year time period 11 infants were identified with permanent hearing loss, resulting in an impairment rate of 2 per 1000. The mean age of hearing loss confirmation decreased from 8.7 months to 3.5 months, and the age at amplification declined from 13.3 months to 5.7 months. CONCLUSION: We conclude that time and experience are important factors in the development and refinement of a universal hearing screen program. Hearing screen outcome data collected over a 4-year period in Rhode Island reveal a steady improvement in the percent of infants completing the 2-stage screen process, the stage 1 and stage 2 refer rates, compliance with rescreen and diagnostic testing, and significant improvement in the age of identification and age of amplification. PMID- 9738716 TI - Persistence of developmental dyscalculia: what counts? Results from a 3-year prospective follow-up study. AB - OBJECTIVE: To study the natural history of developmental dyscalculia (DC), a specific learning disability affecting approximately 5% of the normal school age population and to identify factors that contribute to persistence. STUDY DESIGN: Of a cohort of 3029 fourth-grade students, 185 children were classified as having DC; 140 participated in phase 1 in which they underwent IQ testing; arithmetic, reading, and writing evaluations; and an assessment for attention deficit/hyperactivity disorder over a 3-year period. Three years later (phase 2), 88% of the children (123 of 140) were retested. RESULTS: The arithmetic scores of 95% of the 123 children with DC fell within the lowest quartile for their class. At phase 2, 47% (57 of 123) of the children were reclassified as having persistent DC, scoring in the lowest 5% for their age group (13 to 14 years old). Factors significantly associated with persistence of DC in a multivariate model were severity of the arithmetic disorder and arithmetic problems in siblings of the probands. Factors that were not associated with persistence included socioeconomic status, gender, the presence of another learning disability, and educational interventions. CONCLUSIONS: The outcome of DC is similar to that of other learning disabilities, with a persisting course in almost half of affected children; the remainder continue to perform poorly in arithmetic. The ultimate outcome of children with dyscalculia and the effect on education, employment, and psychologic well-being have yet to be determined. PMID- 9738717 TI - Absence of the fragile X CGG trinucleotide repeat expansion in girls diagnosed with a pervasive developmental disorder. AB - The purpose of this study was to determine the prevalence of the fragile X (FRAX) CGG trinucleotide expansion in a population of young girls (n = 45) diagnosed with pervasive developmental disorder (PDD). Their mean age was 43.7 months (range, 25 to 132 months). Diagnoses included autistic disorder (n = 20), PDD (n = 23), and Asperger's syndrome (n = 2). Molecular FRAX testing was performed on all patients by using the Southern gene blot technique. Genomic DNA was digested with both EcoRI and EagI, fractionated on agarose gel, and blotted and probed with the radiolabeled StB12.3 FMR-1 probe. None of the subjects were found to have an expansion of CGG in either the 2.8 kb or 5.2 kb fragments. A 95% CI, for the prevalence of the FRAX mutation in female subjects with PDD, has an upper bound of 2.9%. We conclude that the prevalence of FRAX positivity in girls with PDD is lower than previously reported. This raises the question of whether any association between FRAX and PDD in female subjects is specific to PDD or is related rather to the presence of mental retardation. PMID- 9738718 TI - Behavior change after growth hormone treatment of children with short stature. AB - OBJECTIVES: To measure the prevalence of behavioral and learning problems among children with short stature and to assess the effect of growth hormone (GH) treatment on such problems. STUDY DESIGN: A total of 195 children with short stature (age range 5 to 16 years, mean age 11.2 years) were tested for intelligence, academic achievement, social competence, and behavior problems before beginning GH therapy and yearly during 3 years of treatment. Children were classified as having growth hormone deficiency (GHD) when GH responses to provocative stimuli were <10 ng/mL (n = 109) and as having idiopathic short stature (ISS) when >10 ng/mL (n = 86). A normal-statured matched comparison group was tested at the baseline only. RESULTS: Seventy-two children in the GHD group and 59 children in the ISS group completed 3 years of GH therapy and psychometric testing. Mean IQs of the children with short stature were near average. IQs and achievement scores did not change with GH therapy. Child Behavior Checklist scores for total behavior problems were higher (P < .001) in the children with short stature than in the normal-statured children. After 3 years of GH therapy these scores were improved in patients with GHD (P < .001) and ISS (P < .003). Also, there was improvement in the scores of children in the GHD group in the internalizing subscales (withdrawn: P < .007; somatic complications, P < .001; anxious/depressed, P < .001) and on the 3 components of the ungrouped subscales (attention, social problems, and thought problems, each P = .001). Larger effects were observed in the GHD group than in the ISS group. CONCLUSIONS: Many referred children with short stature have problems in behavior, some of which ameliorate during treatment with GH. PMID- 9738719 TI - Hepatitis B surface antigenemia at birth: a long-term follow-up study. AB - OBJECTIVE: To investigate the prevalence and outcome of hepatitis B surface antigenemia in newborns of hepatitis B e antigen (HBeAg)-positive hepatitis B surface antigen (HBsAg) carrier mothers under the current immunoprophylaxis program. STUDY DESIGN: From 1984 to 1993, 665 high-risk newborns born to HBeAg positive HBsAg carrier mothers were prospectively recruited. The newborns were tested for HBsAg soon after birth, before hepatitis B immune globulin administration. All newborns received hepatitis B immune globulin within 24 hours after birth plus subsequent hepatitis B vaccination. Those who were seropositive for HBsAg at birth were regularly followed up for their hepatitis B virus (HBV) markers, liver function profiles, and alpha-fetoprotein levels from 1984 to 1996. RESULTS: Sixteen (2.4%) of the 665 subjects were found to be seropositive for HBsAg at birth, and all remained HBsAg-positive at 6 months of age. Twelve of the 16 received long-term follow-up care, and all were confirmed to have chronic HBV infection. Of the 12, 2 had HBeAg seroconversion, and 1 had alanine aminotransferase flares without HBeAg seroconversion. Delayed appearance of hepatitis B core antibody (anti-HBc) occurred in 2 without alanine aminotransferase elevation. CONCLUSIONS: Current immunoprophylaxis strategy does not protect newborns with surface antigenemia, apparently acquired in utero, from becoming HBV carriers. Immunologic attempts to eliminate HBV may occur in carrier children infected in utero, despite their profound immune tolerance to HBV. PMID- 9738720 TI - Changing epidemiologic pattern of chronic hepatitis C virus infection in Italian children. AB - OBJECTIVE: To evaluate the epidemiologic features of chronic hepatitis C virus (HCV) infection in children. STUDY DESIGN: All 106 children with chronic HCV infection consecutively observed in 3 Italian pediatric centers between 1991 and 1997 entered the study. RESULTS: Fifteen children had a history of non-A, non-B hepatitis, and 5 complained of nonspecific symptoms. The 86 remaining patients were free of symptoms and were recruited after HCV screening for intercurrent diseases, maternal infection, or other putative exposure; 39% (none of 30 children born after 1990) had received transfusions, whereas 44%, had a mother with HCV infection. Of the 47 infected mothers, 36% were or had been intravenous drug users, 15% had received transfusions, and 45% had no history of exposure. CONCLUSIONS: Children with chronic HCV infection are often free of symptoms, and thus HCV screening for putative risk has greatly increased the chances of diagnosis. Vertical transmission seems to now be the most common route of infection. Both current and past maternal intravenous drug abuse are risk factors for pediatric infection; however, in an area with relatively high prevalence of anti-HCV in the general population such as Italy, a consistent proportion of infectious mothers have no risk factors of HCV exposure. PMID- 9738721 TI - Antibodies to E2 protein of hepatitis G virus in children: different responses according to age at infection. AB - OBJECTIVES: To study viral persistence and antibody responses after hepatitis G virus (HGV) infection in children of various ages. STUDY DESIGN: We performed an enzyme immunoassay for antibodies to E2 protein (anti-E2) of HGV and reverse transcription polymerase chain reaction assay for HGV RNA on serum samples. RESULTS: Of 28 infants born to HGV RNA-positive mothers, 17 were found to be positive for HGV RNA. None were positive for anti-E2. All 17 infected infants continued to have viremia except 1 who converted to HGV RNA-negative status at 24 months. Six infants had mild elevations of alanine aminotransferase levels (5 HGV positive and 1 HGV-negative). An additional 14 HGV-infected children (aged 6 months to 14 years) with posttransfusion HGV infection were tested for anti-E2 3 months and 12 months after blood transfusion. None of the HGV RNA-positive serum samples were positive for anti-E2; however, 4 of the 8 children with resolving HGV infection were positive for anti-E2 1 year later. CONCLUSIONS: Mother-to infant transmission of HGV resulted in a high viral persistence rate and lack of immune responses to HGV. In contrast, anti-E2 appeared in children who recovered from posttransfusion HGV infection. Mode of transmission and age at infection may be important factors in determining persistent HGV infection and defective immune response to HGV. PMID- 9738722 TI - Primary human herpesvirus 7 infection: a comparison of human herpesvirus 7 and human herpesvirus 6 infections in children. AB - OBJECTIVE: To define the clinical and virologic characteristics of primary human herpesvirus 7 (HHV-7) infection and to compare these characteristics with those of primary human herpesvirus 6 (HHV-6) infection. STUDY DESIGN: A prospective convenience sample study of 496 children < or =3 years old. HHV-7 and HHV-6 infections were identified by viral isolation. Polymerase chain reaction and serology for HHV-7 and HHV-6 were performed. Clinical and laboratory characteristics of patients were obtained from medical records and follow-up interviews. RESULTS: Children with primary HHV-7 infection (n = 8) were identified and compared with children with primary HHV-6 infection (n = 29) detected during the same time period. All children were febrile (mean temperature 39.8 degrees C) with no difference in the degree of fever, frequency of rash, or gastrointestinal complications between the groups. The median age of children with primary HHV-7 infection was 26 months, significantly older than that of children with primary HHV-6 infection (median, 9 months). Children with primary HHV-7 infection were also more likely than those with primary HHV-6 infection to have seizures associated with the illness (P = .004). CONCLUSION: Primary infection with HHV-7 can cause a highly febrile illness in childhood, complicated by seizures. The serologic diagnosis of primary HHV-6 and HHV-7 infections may be confounded by cross-reacting antibodies. PMID- 9738723 TI - Polymerase chain reaction-based detection of rhinovirus, respiratory syncytial virus, and coronavirus in otitis media with effusion. AB - OBJECTIVES: To study the association of human rhinovirus (HRV), respiratory syncytial virus (RSV), and human coronavirus infections in children aged 6 months to 12 years with otitis media with effusion (OME). To determine how long HRV RNA can be detected after HRV infection. METHODS: Middle ear effusion (MEE) samples collected at the time of tympanostomy tube placement from 100 children with OME were examined. Viral RNA was detected by reverse-transcriptase polymerase chain reaction. For HRV the results were compared with virus isolation in cell culture. In vitro studies of the persistence of HRV infectivity and RNA were conducted by combining approximately 10(5) median cell culture infectious doses of HRV with pooled MEE at 37 degrees C and assaying serial samples for 12 weeks. RESULTS: Virus RNA was detected in 30 children. HRV was detected by reverse-transcriptase polymerase chain reaction in 19 children with OME and by virus isolation in 5 children. RSV RNA was found in 8 and HCV in 3 children with OME. No dual viral infection was found. Bacterial pathogens were isolated from 35 MEE samples and were associated with viral RNA in 11 cases, most often with HRV (9 cases). Under in vitro conditions, HRV culture positivity declined rapidly (<2 days), but RNA was detectable for up to 8 weeks. CONCLUSIONS: These results suggest that virus infection, particularly HRV infection, either alone or concurrent with bacteria, is present in a larger percentage of children with OME than previously suspected. It remains to be determined how often the presence of viral RNA in MEE represents persistent RNA, ongoing viral replication, or recurrent infection. PMID- 9738724 TI - Side effects of 2 different dexamethasone courses for preterm infants at risk of chronic lung disease: a randomized trial. AB - OBJECTIVE: We hypothesized that a pulsed course of dexamethasone would result in better linear growth than a 42-day reducing course in preterm infants at risk for chronic lung disease of prematurity. STUDY DESIGN: Forty infants with a birth weight of < or =1,250 g who required mechanical ventilation at 7 days of age were randomly assigned to a repeatable 3-day pulse course of dexamethasone commencing immediately or a 42-day (long) course commencing at 14 days of age if they still required mechanical ventilation and supplemental oxygen. The primary outcome measure was linear growth at 36 weeks' postmenstrual age measured by knemometry. RESULTS: There was no difference in lower leg length at 36 weeks' postmenstrual age. Infants receiving the pulse course had lower rises in blood pressure, less myocardial hypertrophy, and less adrenal suppression. However, more infants required supplemental oxygen at 28 days' postnatal age (14/18 vs 8/21, P < .05) and 36 weeks' PMA (8/16 vs 5/20, P = .12). CONCLUSION: In preterm infants at risk for chronic lung disease, a pulsed course of dexamethasone has fewer side effects than a long course but may be less effective at preventing chronic lung disease. PMID- 9738725 TI - Effects of lactose intake on lactose digestion and colonic fermentation in preterm infants. AB - The purpose of this study was to determine whether doubling the lactose concentration in formula for preterm infants lowers the fraction of lactose digested and/or increases the fraction of lactose fermented. Six preterm infants, 31 to 36 weeks' postconceptional age, were fed a standard preterm formula (carbohydrate is 50% lactose and 50% glucose polymer)(SC) and/or the same volume of formula modified to contain lactose as the sole carbohydrate (LAC). Relative lactose digestion during the LAC formula feeding compared with SC formula feeding was measured by using a stable isotope approach for quantifying the fractional contribution of formula lactose to plasma glucose enrichment. Relative lactose digestion was 0.98 +/- 0.17 (range, 0.70 to 1.19). Fractional fermentation of lactose was estimated from breath H2 excretion (0.52 +/- 0.34 during LAC feeding and 0.23 +/- 0.22 during SC feeding, P = .11). The rate of breath H2 excretion was much higher with LAC (1.34 +/- 0.98 mL/h) than with SC (0.27 +/- 0.29, P = .029). In conclusion, doubling the lactose concentration had only modest effects on fractional lactose digestion. Increased breath H2 excretion with LAC may relate to fermentation of nonlactose sugar or to ill-defined changes in colonic physiology or motility, which could enhance colonic fermentation of malabsorbed sugar by H2-producing bacteria. PMID- 9738726 TI - An optimality score for the neurologic examination of the term newborn. AB - We describe the application of a revised version of the Dubowitz neurologic examination of the newborn in 224 low-risk, term newborn infants. The method has been updated by eliminating less useful items and including new items evaluating general movements and patterns of distribution of tone. An optimality score is included to make the evaluation more quantitative and for comparison with sequential examinations with neurophysiologic and imaging findings. The score is based on the distribution of the scores for each item in the population of low risk term infants. We defined not only the most common pattern for each item but also the variability of the findings by using 10th and 5th centiles. Because most of the items assessing tone and the Moro reflex varied with gestational age between 37 and 42 weeks, the changes were incorporated in the scoring system. The total optimality score was the sum of the optimality scores of individual items. Although the association of 4 or more deviant scores was found in less than 10% of our infants, deviant results on 1 or 2 single items could be observed in a third of this normal population, suggesting that isolated deviant signs have little diagnostic value. In contrast, an abnormal distribution of tone patterns, which we have commonly observed in infants with brain lesions, was not found in this cohort. PMID- 9738727 TI - Familial lipoprotein lipase deficiency in infancy: clinical, biochemical, and molecular study. AB - OBJECTIVES: To describe the characteristics of lipoprotein lipase (LPL)-deficient patients seen in infancy and to evaluate the safety and efficacy of severe fat restriction. METHODS: Children <1 year old presenting with chylomicronemia between 1972 and 1995 were identified, and their clinical courses were reviewed retrospectively. RESULTS: LPL deficiency was demonstrated in 16 infants who presented with irritability (n = 7), lower intestinal bleeding (n = 2), pallor, anemia, or splenomegaly (n = 5), and a family history or fortuitous discovery (n = 2). All plasma samples were lactescent at presentation. Chylomicronemia responded rapidly to dietary fat restriction, and it was possible to maintain satisfactory metabolic control for a prolonged period of time. Only 1 adolescent girl had an episode of pancreatitis associated with the use of oral contraceptives. No persistent adverse effects on growth were seen. We obtained abnormal values for serum iron, alkaline phosphatase, and total calcium. CONCLUSIONS: The presentation of LPL deficiency is heterogeneous during infancy. Close dietary monitoring is required to avoid nutritional deficiencies. Estrogen therapy should be avoided in LPL-deficient patients. PMID- 9738728 TI - Montelukast once daily inhibits exercise-induced bronchoconstriction in 6- to 14 year-old children with asthma. AB - OBJECTIVE: To determine whether montelukast, a leukotriene receptor antagonist, attenuates exercise-induced bronchoconstriction (EIB) in 6- to 14-year-old children with asthma. STUDY DESIGN: Double-blind, multicenter, 2-period crossover study. Children (n = 27) with forced expiratory volume in 1 second (FEV1) > or =70% of the predicted value and a fall in FEV1 > or =20% after exercise on 2 occasions. Patients received montelukast (5-mg chewable tablet) or placebo once daily in the evening for 2 days in crossover fashion (at least 4 days between treatment periods). Standardized exercise challenges were performed 20 to 24 hours after the last dose in each period. End points included area above the postexercise percent fall in FEV1 versus time curve (AAC0-60 min), maximum percent fall in FEV1 from pre-exercise baseline, and time to recovery of FEV1 to within 5% of pre-exercise baseline. RESULTS: Montelukast significantly reduced AAC0-60 min (265 vs 590% x min for montelukast and placebo, respectively, P < or = .05; approximately 59% protection relative to placebo) and the maximum percent fall (18% vs 26% for montelukast and placebo, respectively, P < or = .05). Montelukast treatment resulted in a shorter time to recovery (18 vs 28 minutes for montelukast and placebo, respectively, P = .079). CONCLUSIONS: Montelukast attenuates EIB at the end of the dosing interval in 6- to 14-year-old children with asthma. PMID- 9738729 TI - Lower glycemic response to sucrose in the diets of children with type 1 diabetes. AB - OBJECTIVE: To compare glycemic responses of isocaloric mixed meals containing 2% and 17% sucrose in children with type 1 diabetes who had fasting euglycemia. STUDY DESIGN: Nine children (11 to 16 years) with type 1 diabetes were randomized in a crossover design to receive 2 isocaloric diets (2% or 17% sucrose) in the Clinical Research Center. In the 2% sucrose diet, starch isocalorically replaced sucrose. RESULTS: Fasting euglycemia was comparable on both study days (mean +/- SEM: 2% sucrose, 5.0 +/- 0.3 mmol/L or 90 +/- 5 mg/dL; 17% sucrose, 5.0 +/- 0.3 mmol/L or 91 +/- 6 mg/dL). The 17% sucrose diet resulted in a lower glycemic response than the 2% sucrose diet over the 4-hour study period (area under glucose response curve: mean +/- SEM, 37 +/- 3.5 mmol/L x 4 h vs 42 +/- 4.7 mmol/L x 4 h, P = .01). Peak blood glucose response was earlier and lower (2.2 to 2.8 mmol/L, 40 to 50 mg/dL) with the 17% sucrose diet. CONCLUSIONS: Sucrose in moderate amounts, isocalorically exchanged for starch, lowered glycemic response between breakfast and lunch in children who were euglycemic before breakfast. These data refute concerns about adverse glycemic effects of sucrose and support the use of sucrose-containing foods in the context of a healthy meal plan. PMID- 9738730 TI - Catch-up growth occurs after renal transplantation in children of pubertal age. AB - OBJECTIVE: Assessment of growth after renal transplantation in children of pubertal age by analyzing the annual increment in height standard deviation score (Ht SDS) in all girls > or =10 years and boys > or =11 years of age at the time of transplantation until latest follow-up (minimum 2 years). PATIENTS: A total of 59 grafts were placed in 54 recipients (30 boys) between December 1984 and January 1995. Mean (range) age at transplantation was 13.6 years (10.1 to 17.7 years). Fifty-one percent had congenital renal disease, 36% acquired renal disease, and 13% had hereditary nephropathies. Eighty-seven percent were first grafts; of these, 29% were performed pre-emptively, and 23% were from living related donors. RESULTS: Mean (SD) Ht SDS at transplantation was -1.8 (0.2) and increased significantly thereafter, such that it was -1.6 (0.2) at 1 year, n = 52; -1.5 (0.2) at 2 years, n = 47; -1.0 (0.2) at 3 years, n = 27; -0.7 (0.3) at 4 years, n = 19; and -0.6 (0.3), n = 13, at 5 years after transplantation (analysis of variance, P < .001). The greatest improvement in Ht SDS in the first year was seen in children with the highest glomerular filtration rate (r = 0.429, P = .002) and in those who were shortest at the time of transplantation (r = -0.356, P = .009). CONCLUSION: Catch-up growth occurs in children receiving renal transplants during the expected time of puberty. PMID- 9738732 TI - Sciatica as a manifestation of idiopathic megacolon: a previously undescribed causal relationship. PMID- 9738731 TI - Costello syndrome: phenotype, natural history, differential diagnosis, and possible cause. AB - We describe 8 patients affected with Costello syndrome including an affected sib pair and review the literature on 29 previously reported cases. We emphasize an association with advanced parental age, which is consistent with autosomal dominant inheritance with germline mosaicism. The pathogenesis appears to involve metabolic dysfunction, with growth disturbance, storage disorder appearance, acanthosis nigricans, hypertrophic cardiomyopathy, and occasional abnormalities of glucose metabolism. Although the cause is currently unknown, Costello syndrome is interesting because of a potential genetic-metabolic etiology. PMID- 9738733 TI - Familial granulomatous arthritis (Blau syndrome) with granulomatous renal lesions. AB - Blau syndrome is a granulomatous disease of the skin, eyes, and joints, usually without visceral involvement. It is inherited in a autosomal dominant manner. The Blau susceptibility locus has been mapped to chromosome 16 p 12-q21. A recent report has added liver granulomata. We describe a family with Blau syndrome in whom 1 member had renal interstitial granulomata. PMID- 9738734 TI - Acute hemodynamic effects of pulsed delivery of low flow nasal nitric oxide in children with pulmonary hypertension. AB - We studied 8 children, ages 8 months to 14 years, during cardiac catheterization in order to determine the acute hemodynamic effects of pulsed nasal cannula delivery of nitric oxide (NO) in children with pulmonary hypertension. NO was administered by continuous mask or pulsed nasal cannula in random order. All patients effectively triggered the NO pulsing device. Pulsed delivery of inhaled NO lowered mean pulmonary artery pressure and pulmonary vascular resistance as effectively as mask delivery of NO. Pulsed inhaled NO delivery may potentially be useful for the long-term domiciliary treatment of pulmonary hypertension in children. PMID- 9738735 TI - A 7-week-old boy with respiratory congestion. PMID- 9738736 TI - Cutaneous manifestations of disseminated fungal infection in an immunocompromised child. PMID- 9738738 TI - Plastibell circumcision: a novel complication. PMID- 9738737 TI - Barrier function of neonatal skin. PMID- 9738739 TI - Photodynamic therapy and the treatment of head and neck neoplasia. AB - OBJECTIVE: To present the theory, technique, and results of photodynamic therapy for the treatment of oral, laryngeal, and head and neck cancers. STUDY DESIGN: Retrospective review of the literature of more than 500 patients with head and neck cancer treated with photodynamic therapy, as well as a retrospective review of the author's 107 patients treated with photodynamic therapy for head and neck neoplasia between 1990 and 1997. METHODS: The literature was retrospectively reviewed, as were patient records, and tabulated for age, sex, site, and staging of lesions, with special focus on post-photodynamic therapy treatment outcome, long-term disease-free survival, and complications. RESULTS: Twenty-five patients with carcinoma in situ and T1 squamous cell carcinoma of the true vocal cord who underwent photodynamic therapy treatment for cure obtained a complete response after a single photodynamic therapy treatment. Only one patient has had recurrence to date, with a cure rate to 79-month follow-up of 95%. Twenty-nine patients with carcinoma in situ and T1 recurrent squamous cell carcinomas of the oral cavity and tongue were treated. All obtained a complete response after a single photodynamic therapy treatment; however, five patients developed local recurrence with follow-up to 70 months, for an 80% cure rate. A review of 217 patients with early squamous cell carcinomas of the head and neck treated with photodynamic therapy in the literature demonstrated an 89.5% complete response rate. The most common complication in these patients was limited prolonged skin photosensitivity without any permanent sequelae. CONCLUSIONS: Photodynamic therapy is effective for treating carcinoma in situ and T1 squamous cell carcinoma of the larynx and oral cavity and may be of benefit as an adjuvant intraoperative treatment of stages III and IV tumors of the head and neck in conjunction with surgery and radiation therapy to improve cure rates. Further controlled studies need to be performed to further demonstrate the effectiveness of photodynamic therapy and the treatment of head and neck cancers. PMID- 9738740 TI - Laser and cisplatinum for treatment of human squamous cell carcinoma. AB - OBJECTIVE: Interstitial laser therapy (ILT) with the neodymium:yttrium-aluminum garnet (Nd:YAG) (1064 nm) laser via fiberoptics is becoming a more precise, minimally invasive alternative for thermoablation of unresectable or recurrent head and neck neoplasms, but recurrence is often seen at the margin. Combining intratumor chemotherapy with interstitial laser should be most effective using drugs activated by thermal energy. The objective of the current study was to test intratumor cisplatinum (cis-diaminedichloroplatinum [CDDP]) injections given in conjunction with laser therapy as an experimental approach for improved treatment of squamous cell carcinoma (SCC). METHODS: Human SCC tumors were grown as subcutaneous transplants in nude mice and injected with CDDP (0.4 to 1.2 mg/g) in water or in collagen-based gel carrier with epinephrine (epi-gel) followed by ILT via 0.6-mm fiberoptics coupled to an Nd:YAG laser (1064 nm/180 J). RESULTS: Tumors injected with CDDP epi-gel exhibited a partial response with two- to fourfold tumor delay compared with aqueous drug or untreated SCC transplants during 10 weeks' follow-up. Combined drug and laser therapy significantly (P < .01) decreased tumor volume, with recurrence in only 25% of animals tested compared with 78% tumor regrowth after ILT alone. CONCLUSION: These initial results suggest that laser chemotherapy may become an effective treatment for advanced head and neck cancer. PMID- 9738741 TI - Palliative laser therapy for recurrent head and neck cancer: a Phase II clinical study. AB - OBJECTIVES: Laser therapy is becoming a more precise, minimally invasive alternative for tumor ablation. Recent reports confirm successful palliation of pain and functional disabilities in patients with advanced deep carcinoma of the head and neck using interstitial laser phototherapy (ILT). STUDY DESIGN, PATIENTS, AND METHODS: The current study describes an ongoing Phase II trial of neodymium/yttrium-aluminum-garnet (Nd:YAG) laser therapy for palliation of advanced head and neck cancer. A total of 40 advanced cancer patients have been entered into this protocol (25 men and 15 women). RESULTS: Nineteen of these patients had no evidence of recurrence after ILT with an average follow-up of 11 months (range, 2 to 24 mo). Currently, 19 of these patients are alive, 14 with tumor remission and six with persistent disease. A total of 79 tumor sites received ILT with 43 (54.5%) completely ablated. Stratified by tumor site, ILT led to a complete response in 21 of 24 in the oral cavity, eight of 28 neck tumors, four of 10 in skin, and 10 of 17 in other sites. The procedure was well tolerated in most cases and was repeated at intervals in patients with residual disease or recurrences for a total of 118 laser treatments (average, 2.95 treatments per patient). CONCLUSIONS: The results suggest that ILT can be performed safely and repeated as needed, and may be less costly than conventional surgery for head and neck cancer. However, additional follow-up is needed to obtain convincing evidence of long-term therapeutic benefits. PMID- 9738742 TI - Dose-related tissue effects of the CO2 and noncontact Nd:YAG lasers in the canine glottis. AB - OBJECTIVES: The CO2 laser is the standard for control of recurrent respiratory papillomatosis because of its predictable action on laryngeal tissue. The noncontact neodymium:yttrium aluminum garnet (Nd:YAG) 1064-nm laser is generally not used in the larynx owing to the lack of data on its tissue effects, and its potential lack of safety in the larynx. Combined Nd:YAG and CO2 laser treatments have been used safely in the tracheobronchial tree to eradicate recurrent respiratory papillomas. The objectives of this study were to describe and evaluate a method for applying the noncontact Nd:YAG laser to the larynx, to compare the tissue effects of the Nd:YAG, CO2, and combined Nd:YAG and CO2 lasers in the canine larynx, and to extrapolate canine tissue data to the human. METHODS: The CO2, Nd:YAG, and combined Nd:YAG/CO2 lasers were applied to the glottis in four mongrel dogs. Laryngectomy was performed and the tissue was examined histologically. The nature and degree of tissue injury were analyzed relative to laser type and energy data. RESULTS: In the canine larynx, the CO2 laser vaporized the surface epithelium and caused varying degrees of edema and necrosis of the lamina propria. The Nd:YAG laser did not cause ulceration but did show a greater degree of thermal damage to the lamina propria. Combined Nd:YAG/CO2 applications resulted in separation of the perimysial fibers from the muscle fibers of the vocalis muscle. CONCLUSION: These findings suggest that the noncontact Nd:YAG laser can be applied in a controlled fashion to the canine larynx at appropriate power densities. Anatomical differences between human and canine larynges are considered. Extrapolation to humans is proposed. PMID- 9738743 TI - Hyperbaric oxygen for the management of radionecrosis of bone and cartilage. AB - OBJECTIVES: To review the use of hyperbaric oxygen in the management of radionecrosis of the head and neck. STUDY DESIGN: A retrospective analysis of patients utilizing chart review and telephone interviews. All patients diagnosed with osteoradionecrosis and chondroradionecrosis of the head and neck and treated with hyperbaric oxygen at the University of Virginia are included. METHODS: Demographics, pretreatment data, and precipitating events were recorded. Outcomes were evaluated using a grading scale of symptomatology and physical examination as determined by the patient and physician. RESULTS: Sixteen patients with osteoradionecrosis and five with chondroradionecrosis were reviewed. All patients showed clinical improvement with decreased pain following HBO therapy. None of the patients with chondroradionecrosis required laryngectomies, and two of the four who were tracheotomy dependent were successfully decannulated. The patient and physician grading scores demonstrated moderate to significant improvement in both groups following therapy. CONCLUSION: The successful use of hyperbaric oxygen for the management of radionecrosis of the head and neck is supported. The unusual prevalence of chondroradionecrosis may be an early reflection of changes in treatment protocols for patients with head and neck cancer. PMID- 9738744 TI - Adenoid cystic carcinoma of the trachea. AB - OBJECTIVE: Primary tracheal tumors are rare, occurring in 0.2 per 100,000 persons per year. Adenoid cystic carcinoma (ACC) is the second most common histologic type of tracheal malignancy. Its clinical behavior is different from the other tracheal neoplasms and thus should be studied separately. STUDY DESIGN/METHODS: Retrospective review of the medical records of six patients with tracheal ACC who were treated at University Hospitals of Cleveland between 1971 and 1996 and literature review. RESULTS/CONCLUSION: Tracheal ACC is an indolent tumor that affects people at any age but has a peak incidence in the fifth decade. There is a nearly equal male-to-female ratio. Almost half of tracheal ACCs occur in the proximal trachea, accounting for the most common presenting symptoms: dyspnea, cough, and hoarseness. Because of the hoarseness, patients are often referred to an otolaryngologist. Complete resection provides the best chance for increased survival. Neutron beam radiotherapy holds promise for adjuvant therapy. PMID- 9738745 TI - Paradoxical spread of renal cell carcinoma to the head and neck. AB - OBJECTIVES: To present cases of renal cell carcinoma presenting with only head and neck metastases, to review theories of physiology and anatomy describing this phenomenon, and to discuss the role of the otolaryngologist in the treatment of these lesions. STUDY DESIGN: Retrospective review of the records of three patients who presented with renal cell carcinoma with head and neck metastases over the 3-year period from 1992 to 1995. METHODS: Retrospective review of the records of three patients who presented with renal cell carcinoma with head and neck metastases. In addition, English-language literature was reviewed with special focus on the anatomic and physiologic pathways possible to allow for such a phenomenon. CONCLUSIONS: Renal cell carcinoma has an occasional presentation as a head and neck mass without evidence of disease elsewhere. Various routes of spread have been postulated. Batson's venous plexus, as postulated by Nahum and Bailey, is an anatomic route through which emboli could navigate to the head and neck and avoid pulmonary vascular filtration. Interactions on the cellular level may also be responsible for the seemingly paradoxical spread. We recommend local excision of head and neck metastases of renal cell carcinoma without sacrifice of vital structures as a sound treatment regimen. PMID- 9738747 TI - The agony of nasal polyps and the terror of their removal 200 years ago: one surgeon's description. AB - In previous times, nasal polyps could grow to enormous size, especially in cases of incomplete removal. Many patients suffered a lingering death from the disease or a faster one from surgical attempts at removal. John Bell's operative procedure for this condition is reviewed, showing how terrifying it must have been for the patient. PMID- 9738746 TI - Relationships between otitis media sequelae and age. AB - OBJECTIVES: To explore relationships between age and sequelae in two groups of children treated with tympanostomy tubes for chronic otitis media with effusion (OME). STUDY DESIGN: Cross-sectional study of sequelae among children, adolescents, and adults at 4 years and 9 to 23 years after tympanostomy tube treatment. METHODS: Group I was examined with otomicroscopy, tympanometry, and audiometry two to four times a year as part of a prospective study, and they were evaluated 4 years after initial tube treatment for this study. Group II received tubes while participating in a chronic OME study, but participants were not followed prospectively after treatment. Nine to 23 years after tube treatment, they were examined with otomicroscopy, tympanometry, and hearing screening. RESULTS: Among the 5- to 28- year-old subjects, cholesteatoma (< or = 1%) and perforation (< or = 2%) were rare. In Group I, tympanosclerosis increased with age (P < .01), and OME (flat tympanograms) decreased with age in Group II (P < .01). The older cohort was more likely to have severe retractions (18% vs. 4%, P = .02), hearing loss (21% vs. 10%, P < .01), and severe atrophy (24% vs. 0%, P < .01) than the younger cohort, but they were less likely to have flat tympanograms (2% vs. 12%, P < .01). CONCLUSIONS: Although OME became less prevalent with age, important sequelae (severe atrophy, severe tympanic membrane retraction, hearing loss, cholesteatoma, and chronic perforation) may develop in children with chronic OME as they become adolescents and young adults. Long-term prospective studies are important in defining the progression of sequelae in these children. PMID- 9738748 TI - Sinus lateralis in endoscopic ethmoidectomy. AB - OBJECTIVE: To characterize the anatomy of the sinus lateralis and enable a more accurate and safe approach to endoscopic ethmoidectomy. STUDY DESIGN: An anatomic study of 33 cadaver heads providing a prospective evaluation of 50 ethmoid sinuses. The sinus lateralis, in particular, was thoroughly examined and typical variations were classified into four categories. A technique using the sinus lateralis as a critical landmark is described which allows calculated removal of anterior ethmoid cells in a posterior-to-anterior direction, avoiding inadvertent entry into the posterior ethmoid. Prospective evaluation of this technique in 12 pediatric patients found it to be safe and particularly useful for neophyte endoscopic surgeons. METHODS: Endoscopic examination of 50 cadaver sinuses using 0-, 30-, and 70-degree, 4-mm telescopes to dissect the anterior ethmoid. The characteristics of each sinus lateralis were documented. To confirm the endoscopic findings, we grossly examined each specimen. RESULTS: Four different categories of sinus lateralis formation were identified: type I (n = 22), posterosuperior extension to skull base; type II (n = 15), posterior extension to sphenoid face; type III (n = 8), abrupt termination posterior to ethmoid bulla; and type IV (n = 5), extension into posterior ethmoid through dehiscent basal lamella. CONCLUSIONS: The sinus lateralis is a consistent feature of the anterior ethmoid. Type I and II patterns are most conducive to the aforementioned technique. Type III is the most difficult to identify endoscopically, whereas type IV is most apt to encourage an unplanned posterior ethmoidectomy. Regardless of the chosen ethmoidectomy technique, careful assessment of the sinus lateralis should enable more accurate and safe removal of ethmoid disease with reduced complications. PMID- 9738749 TI - An objective analysis of the impact of lateral rhinotomy and medial maxillectomy on nasal airway function. AB - OBJECTIVE: The lateral rhinotomy and medial maxillectomy procedure, while known to interrupt nasal valve supports, has not previously been reported to adversely affect nasal airway function. The purpose of this study was to utilize state-of the-art techniques to objectively analyze the impact of this procedure on nasal airway function. DESIGN: The study design was retrospective and subject controlled. METHODS: The study population was derived from an academic, tertiary referral, otolaryngology-head and neck surgery department with an estimated catchment population of 4 million people. Subjects included 21 consecutive, long term postoperative patients who had undergone lateral rhinotomy and medial maxillectomy for inverted papilloma. Objective measures included vestibular cephalometric measurements, airflow rhinomanometry, and acoustic rhinometry. RESULTS: Statistically significant results reveal that although lateral rhinotomy and medial maxillectomy are associated with alar collapse, both overall nasal airflow and valve areas are increased. CONCLUSION: Lateral rhinotomy and medial maxillectomy does not adversely affect nasal airway function. This appears to be the result of concomitant resection of the functionally dominant inferior turbinate. This suggests that lateral rhinotomy performed in conjunction with operations not requiring inferior turbinectomy, such as anterior craniofacial resection, may adversely affect nasal airway function. PMID- 9738750 TI - Efficacy of tonsillectomy for recurrent throat infection in adults. AB - OBJECTIVE/HYPOTHESIS: Sore throats result in health care visits, use of oral antibiotics, and days off work or school for many patients who do not meet American Academy of Otolaryngology-Head and Neck Surgery (AAO-HNS) guidelines for tonsillectomy. We sought to determine whether tonsillectomy would benefit this group. STUDY DESIGN: Retrospective analysis of the medical records of all patients aged 16 years or older who had tonsillectomy at our institution between 1988 and 1993. METHODS: Number of clinic visits, number of throat cultures positive for streptococci, and number of prescriptions for oral antibiotics recorded for 147 patients during the 2-year periods before and after tonsillectomy were tabulated. Statistical comparisons were made using the Student's t test. Mean number of clinic visits and oral antibiotics prescribed for throat infection before tonsillectomy were significantly higher than after tonsillectomy. RESULTS: Patients who had throat cultures positive for streptococci had more preoperative clinic visits and use of oral antibiotics than patients whose throat cultures were not positive for streptococci. When surveyed by telephone, most (>87%) of the respondents reported that they had fewer and less severe sore throats, required fewer days off work or school, and would recommend the procedure. CONCLUSIONS: Our results suggest that early tonsillectomy in patients with recurrent throat infection may result in improved patient satisfaction, better health, and improved utilization of medical resources. PMID- 9738751 TI - Spectral analysis of photoplethysmograms from radial forearm free flaps. AB - OBJECTIVE: Photoplethysmography utilizes a green-light-emitting diode to transmit light into a tissue. Reflected light from hemoglobin in dermal capillary red blood cells is received by a photo detector and is analyzed as light intensity along a frequency spectrum. This method of analysis allows for the removal of "noise" above (stray light and alternating current [AC]) and below (room vibrations and respiratory motion) the peak signal (1 to 2 Hz) and results in a means to distinguish between perfused and nonperfused tissues. METHODS: Twenty two of 30 consecutive radial forearm free flap (RFFF) patients were enrolled in an approved human studies protocol to collect descriptive data for RFFFs that were perfused, arterial occluded, and venous occluded. The protocol was performed following completion of flap elevation and prior to pedicle ligation, flap inset, and microvascular anastomoses. Six 90-second measurements per flap were obtained (n = 132), processed by fast Fourier transform (FFT), and analyzed by blinded reviewers to determine their state of perfusion. Signal was collected 5 minutes after the onset or release of individual vessel occlusion. RESULTS: The reviewers' interpretations were compared with the status of the pedicle and analyzed for sensitivity (0.96), specificity (0.95), and positive predictive value (0.98). CONCLUSIONS: FFT analysis of photoplethysmograms from RFFF patients provides an accurate and rapid means for determining RFFF pedicle vessel patency. Photoplethysmography may provide a clinically useful tool for postoperative perfusion monitoring of free flaps in the future. PMID- 9738752 TI - Prophylactic antibiotics in surgery for chronic ear disease. AB - OBJECTIVES/HYPOTHESIS: The role of prophylactic antibiotics in otologic surgery continues to be debated and perhaps misused. Prior studies have provided conflicting evidence with regard to the benefit obtained from the use of prophylactic antibiotics in surgery for chronic otitis media. The current study was designed to evaluate the role of prophylactic antibiotics in the outcomes of surgery for chronic ear disease. It was the authors' impression that there was no indication for prophylactic antibiotics in such surgery. STUDY DESIGN: Randomized prospective study performed in a tertiary care facility. METHODS: Patients who met inclusion criteria (n = 146) were randomly assigned to an antibiotic treatment group or a control group receiving no prophylactic antibiotics. Patients in the antibiotic treatment group were given preoperative intravenous antibiotics followed by oral antibiotics for 5 days after surgery. Patients were followed postoperatively and observed for clinical evidence of infection and graft failure. RESULTS: There was no statistically significant difference between the two groups with regard to the incidence of postoperative infection or graft survival. CONCLUSIONS: The use of prophylactic antibiotics in surgery for chronic ear disease cannot be recommended based on the findings of this study. PMID- 9738753 TI - Direct bonded orthodontic brackets for maxillomandibular fixation. AB - OBJECTIVES/HYPOTHESIS: Mandibular fracture treatment often includes arch bar maxillomandibular fixation (MMF), either alone or in combination with open reduction/internal fixation (ORIF) techniques. The glove perforation rate associated with arch bar placement, the incidence of blood-borne pathogen positivity in facial fracture patients, and the injurious effects of arch bars on dental enamel and gingiva have prompted the development of safer alternatives to arch bar MMF. This study evaluates the efficacy, ease of use, and safety profile of one such alternative: orthodontic direct bonded bracket fixation (MMF/DBB). STUDY DESIGN: Prospective study of consecutive mandible fracture patients treated with MMF/DBB. METHODS: Thirty-two patients with mandibular fractures were evaluated from January 1994 to July 1997. Fourteen were appropriate for treatment with MMF/DBB (12 men and two woman; mean age, 24.6+/-7.2 y; range, 16-42 y). Fracture sites included symphysis, angle, condylar neck, coronoid, and body. Nine patients underwent MMF/DBB alone; five underwent MMF/DBB with subsequent ORIF. RESULTS: No infection, malocclusion, malunion/nonunion, or enamel/ gingiva injury occurred. Mean follow-up was 6 months (range, 1-12 mo). Oral hygiene with MMF/DBB was superior to historical controls using arch bars. CONCLUSIONS: MMF/DBB can serve as the single treatment method with satisfactory results in patients with favorable, less complicated mandible fractures, although with increased experience, we have treated several more complex cases with MMF/DBB alone. In cases necessitating ORIF, MMF/DBB can be performed preoperatively to align fracture segments and reestablish occlusion. This facilitates placement of osteosynthesis plates and reduces ORIF operative time. MMF/DBB is an economical, safe technique that minimizes blood-borne-pathogen risk to the operative team, eliminates periodontal injury, facilitates postoperative dental hygiene, and is painless to apply and remove. PMID- 9738754 TI - Changing etiology of vocal fold immobility. AB - HYPOTHESIS: Vocal fold immobility is a sign of underlying disease. When the etiology remains unclear, evaluation may become time consuming and costly, and directed work-up imperative. This study examined the hypothesis that the etiologies of vocal fold immobility are changing, with extralaryngeal malignancies and nonthyroidectomy surgical trauma having become more common causes. METHODS: A retrospective review of consecutive patients with vocal fold immobility who had an adequate workup to determine the etiology. RESULTS: Three hundred ninety-seven cases with a determined etiology were identified, yielding 280 unilateral and 117 bilateral immobilities. The largest single category in unilateral immobility was nonlaryngeal malignancy--69 patients (24.7%)--80% of which were pulmonary or mediastinal, followed by 67 patients (23.9%) with immobility secondary to surgical trauma. Thyroidectomy accounted for only 8.2%. The leading cause of bilateral immobility was surgical trauma-30 patients (25.7%) -21 (18%) of whom had thyroidectomy. Acute and chronic intubation injuries accounted for 21 unilateral (7.5%) and 18 bilateral (15.4%) cases. CONCLUSIONS: These data indicate a changing etiology of vocal fold immobility, with growing percentages of extralaryngeal malignancies and surgery-related injuries. These findings have implications for the timing and method of management based on anticipated outcome. PMID- 9738755 TI - The recurrent laryngeal nerve: related vascular anatomy. AB - This study was based on 34 recurrent laryngeal nerve dissections after arterial casting with red-colored latex. The aim was to provide specific information about the perineural microvasculature. This study established the following points: 1. a great anatomic variability does exist; 2. the laryngeal nerve is usually in relation to the posterior branch of the inferior thyroid artery; and 3. this vascular branch is sometimes replaced with a vascular network. In all cases, this microvascularization must be preserved during thyroid surgery. PMID- 9738756 TI - Pharyngo-UES contractile reflex in patients with posterior laryngitis. AB - BACKGROUND: Earlier studies have shown that stimulation of the human pharynx by injection of minute amounts of water stimulates the pharyngo-UES contractile reflex. It has been suggested that this reflex may be activated during pharyngeal reflux of gastric and/or esophageal content, thus increasing the UES pressure and possibly preventing further entry of the refluxate into the pharynx. However, the integrity of this reflex in patients with posterior laryngitis has not been studied. AIM: Evaluate the pharyngo-UES contractile reflex in a group of patients with objective findings of posterior laryngitis. METHODS: Fourteen consecutive patients with posterior laryngitis (mean age, 48+/-6 y) and 13 healthy volunteers (mean age, 53+/-6 y) were studied by concurrent pharyngeal water stimulation and UES manometry. RESULTS: The threshold volume required to evoke the pharyngo-UES contractile reflex in the laryngitis group (0.4+/-0.05 mL) was significantly higher than that of the control (0.2+/-0.04 mL) (P < .05). Following stimulation of the pharyngo-UES contractile reflex, the maximum postinjection pressure in patients (75+/-6 mm Hg) was similar to that of the controls (78+/-6 mm Hg). The percent increase in UES pressure following stimulation of the reflex in the laryngitis group (99%+/-15%) was significantly higher than that of controls (55%+/-11%) (P < .05). CONCLUSIONS: Compared with normal controls, a significantly larger volume of liquid is required to trigger this reflex in patients with posterior laryngitis. When triggered, the maximum UES pressure induced by the pharyngo-UES contractile reflex is similar between the two groups. These findings suggest an altered afferent sensory limb of this reflex in patients with posterior laryngitis. PMID- 9738757 TI - Three-dimensional xenograft model of dysplastic human laryngeal mucosa. AB - OBJECTIVE: Development of new therapeutic interventions in head and neck squamous cell carcinoma (HNSCC) will be facilitated by a model system that incorporates the ease of manipulation found in current tissue culture systems while retaining the three dimensional architecture that defines these malignancies. STUDY DESIGN: Original scientific investigation. METHODS: We describe a modification of a normal respiratory mucosa model system which recreates premalignant mucosal histology. Grossly normal appearing human mucosa is harvested from laryngectomy specimens, the mucosal epithelium selectively removed by protease treatment and placed in conventional tissue culture. After 7 days, the cells are seeded into denuded rat tracheas, which are in turn implanted in flank pockets of athymic nu/nu mice. The tracheas are incubated for three weeks, removed and the mucosa examined histologically. RESULTS: As originally described, normal pseudostratified squamous epithelium can be re-established in this system. Using human dysplastic mucosa as a starting material, mucosal histologies of respiratory dysplasia, squamous metaplasia, squamous dysplasia and squamous carcinoma in situ can be established. CONCLUSION: This system will provide a paradigm for future therapeutic interventions to modify the progression of squamous metaplasia to dysplasia, carcinoma in situ and invasive squamous cell carcinoma. PMID- 9738758 TI - Predicting vocal outcome by means of a vocal endurance test: a 5-year follow-up study in female teachers. AB - OBJECTIVES: Investigate whether vocal problems in future professional activities can be predicted by early laryngeal and phoniatric evaluation and whether a vocal endurance test can contribute to this evaluation. STUDY DESIGN: Five-year follow up study of 30 female education majors, initially documented with a standard voice assessment and a vocal endurance test. Measurements before and after vocal endurance testing were compared and related to the vocal outcome 5 years after the initial testing. METHODS: Voice assessment included perceptual evaluation, airflow measurements, Fo and SPL measurements, voice range profile and laryngeal (stroboscopic) examination. The Standard Tolerance Test, as recommended by the Union of European Phoniatricians, was followed. This data set was completed with a questionnaire concerning the subjects' vocal behavior. This questionnaire was repeated 5 years later. RESULTS: No significant differences were found for ENT scores (laryngostroboscopy) (P = .018). Logistic regression was used to determine a relationship between initial observations and the final outcome. CONCLUSIONS: The role of an endurance test as used in this study is negligible for the prediction of vocal outcome. A combination of laryngeal examination, maximum phonation time, and perceptual evaluation, assessed prior to the endurance test, reveals a prediction of the vocal outcome with a specificity of 90% and a sensitivity of 70%. PMID- 9738759 TI - Familial Mondini dysplasia. AB - OBJECTIVES/HYPOTHESIS: To determine the mode of inheritance of familial nonsyndromic Mondini dysplasia. STUDY DESIGN: Correlative clinical genetic analysis of a single kindred. METHODS: Clinical history, physical examination, audiologic analysis, computed tomography of the temporal bones, and cytogenetic analysis. RESULTS: The male proband, three affected sisters, and an affected brother are offspring of unaffected parents. The mother and an unaffected brother have audiologic findings suggestive of heterozygous carrier status for a recessive hearing loss gene. CONCLUSIONS: Pedigree analysis indicates autosomal recessive inheritance in this family. The observed inheritance and clinical, audiologic, and radiologic findings are different from those previously described for another family with nonsyndromic Mondini dysplasia. The phenotype in this study family therefore represents a distinct subtype, indicating clinical and genetic heterogeneity of this disorder. This information should facilitate future molecular linkage analyses and genetic counselling of patients with inner ear malformations. PMID- 9738760 TI - Auditory neuropathy in childhood. AB - OBJECTIVES: Auditory neuropathy is a recently described disorder in which patients demonstrate hearing loss for pure tones, impaired word discrimination out of proportion to pure tone loss, absent or abnormal auditory brainstem responses, and normal outer hair cell function as measured by otoacoustic emissions and cochlear microphonics. We have identified eight pediatric patients having hearing deficits that are most likely due to a neuropathy of the eighth nerve. In this study, the results of audiologic testing performed with these eight children are described. STUDY DESIGN: Retrospective review of audiologic findings in eight children with auditory neuropathy. METHODS: Each subject was tested with pure tone and speech audiologic testing, auditory brainstem response, and click-evoked otoacoustic emissions. Results of these tests were tabulated and summarized. RESULTS: Pure tone audiologic testing revealed five children with upsloping sensorineural hearing loss, two with high frequency loss, and one with a mild, flat configuration. Six children demonstrated poor word discrimination scores, and the other two had fair to good word discrimination. All eight subjects had normal distortion product and transient otoacoustic emissions. All eight children demonstrated absent or marked abnormalities of brainstem auditory evoked potentials. These findings suggest that while cochlear outer hair cell function is normal, the lesion is located at the eighth nerve. CONCLUSIONS: Recent advances in otoacoustic emissions testing permit differentiation of neural deafness from sensory deafness. This paper describes the clinical presentation and audiologic findings in pediatric auditory neuropathy, as well as the recommended management of these patients. Otolaryngologists should be aware of this disorder and implications for its management, which differs from treatment of sensorineural hearing loss. PMID- 9738761 TI - Embryonic middle ear mesenchyme disappears by redistribution. AB - OBJECTIVE: The fate of embryonic middle ear mesenchyme from a postgestational infant ear has been speculative. Recently a volume analysis of human neonatal temporal bones demonstrated that embryonic mesenchyme disappeared by redistribution and thinning to surface a growing middle ear space. If this model is accurate, interaction with amniotic fluid and the gestational environment should not influence mesenchymal behavior. Therefore an opossum marsupial model was compared with human data. METHODS: The temporal bones of opossum pups (20 to 36 days of age) were sectioned for histologic analysis. Computations were made for the volume of the middle ear air cavity (VAC), volume of the bone cavity (VBC), volume of mesenchyme (VM), and percentage of the middle ear occupied by mesenchyme (%M), which were plotted against height using regression statistics. These data were compared with human neonatal (0 to 30 days of age) temporal bones. RESULTS: In both the opossum and the human the VAC and the VBC increased in parallel with growth of the body. In the opossum the VAC and VBC both grew at 0.148 mm3/mm of body length. In humans, both the VAC and VBC grew 6.1 mm3/cm of body length. The VM in the opossum remained constant at 0.98 mm3, regardless of body length. In humans the VM remained constant at 71.7 mm3, regardless of body length. Therefore the %M proportionately decreased inversely with increasing ear size in both the opossum and the human neonate. CONCLUSION: This study supports a simple and credible explanation for the illusion of mesenchymal disappearance in the neonatal middle ear. The mesenchymal connective tissue redistributes to cover a larger surface area in a persistently enlarging cavity. These findings occur in different species, whether gestation is completed in an intrauterine or an extrauterine environment. PMID- 9738762 TI - Quality of life after acoustic neuroma surgery. AB - OBJECTIVE: To assess how surgery affected the quality of life of patients with acoustic neuromas and to investigate possible predictors of the functional outcome following surgery. STUDY DESIGN: A questionnaire based on the Glasgow Benefit Inventory was completed by patients randomly selected following acoustic neuroma surgery. SETTING: Skull base surgery unit of a university teaching hospital (tertiary referral center). PATIENTS: Fifty-three patients with acoustic tumors (follow-up, 1 to 3 y). RESULTS: With regard to overall quality of life, nine patients (17.4%) reported that it became better, 28 patients (53.8%) worse, and 15 patients (28.8%) that it remained the same. Four patients (7.8%) became better off financially, 15 patients (29.4%) worse, and 32 (62.8%) remained unchanged. Forty-one patients (78.8%) did not change their occupation, and 11 (21.2%) had to change their occupation, mainly because of the adverse effects of the operation. With regard to the age at operation, older patients were found to have better overall quality of life. Moreover, younger patients had worse postoperative financial status and they were more likely to change their occupation after the operation. The tumor size did not significantly affect the overall postoperative quality, but it did affect the postoperative financial status (patients with larger tumors were more likely to have worse postoperative financial status). CONCLUSION: Acoustic neuroma surgery has a significant impact on patients' overall quality of life. Surgeons proposing to operate on small tumors should not assume that the impact on patients' life will be necessarily less than that following the removal of larger tumors. All patients, especially in the younger age group, should be prepared and thoroughly informed about the consequences of the operation on their quality of life. PMID- 9738763 TI - Systemic neutrophil intrinsic 5-lipoxygenase activity and CD18 receptor expression linked to reperfusion injury. AB - OBJECTIVE: To determine if systemic neutrophil intrinsic 5-lipoxygenase (5-LO) inhibition correlates with decreased expression of surface adhesion molecules and attenuation of ischemia-reperfusion (i/r) injury in guinea pig island skin flaps. METHODS: Eighty-one adult female Hartley guinea pigs were divided into one control group, three 2-hour ischemia groups, and four 10-hour ischemia groups. Island dorsal skin flaps were developed (except in the control group), and 2 hours before reperfusion, zileutin (a 5-LO inhibitor) or vehicle was administered orally. Postreperfusion systemic neutrophil receptor expression, neutrophil flap infiltration, and flap survival were measured. Neutrophils from whole blood were analyzed for CD18 containing surface receptor expression using monoclonal antibodies and cell associated fluorescence. Neutrophil infiltration into a distal centimeter squared of flap tissue was assessed using myeloperoxidase antibodies, and flap survival was determined within 7 days postoperatively. RESULTS: Flaps in the treated 2- and 10-hour ischemic groups survived totally intact, while the untreated 10-hour ischemic flaps underwent total necrosis. A significant main effect of the drug was detected using analysis of variance (ANOVA) (P =.0001). Surface receptor detection and neutrophil infiltration were significantly increased in the untreated animals. CONCLUSIONS: Zileuton, a 5-LO inhibitor, reduces adhesion receptor expression on systemic neutrophils and attenuates i/r injury. Systemic neutrophil intrinsic 5-LO activity and CD18 receptor expression are linked to reperfusion injury and may be fundamental events in its pathogenesis. PMID- 9738764 TI - Use of fibrin glue to protect tissue during CO2 laser surgery. AB - OBJECTIVE: Accidental injury of tissues during CO2 laser irradiation can lead to serious morbidity, especially during ear, nose, and throat, neurosurgical, and plastic-reconstructive procedures. This experimental study describes a new technique in which vital structures are coated with a thin layer of fibrin glue to protect them from accidental CO2 laser irradiation. STUDY DESIGN/MATERIALS AND METHODS: The femoral neurovascular bundles (femoral artery, vein, and nerve) of 12 rats were exposed. On one side the bundle was coated with fibrin glue, which is a biological two-component glue consisting of fibrinogen solution and thrombin. Upon application, an elastic mass on the neurovascular bundle was formed. The contralateral neurovascular bundle was left uncoated. Subsequently both bundles were subjected to CO2 laser irradiation at different powers (5, 7, and 9 W), with an irradiation time of 0.1 seconds. Light microscopy was performed at 30 minutes and 2 days after surgery. RESULTS: No macroscopic visible hemorrhages occurred during laser irradiation in the glue-coated bundle. Light microscopic evaluation revealed an undamaged neurovascular bundle without any signs of thermal damage. In the uncoated bundles intraoperative hemorrhages resulting from laser energy occurred in all specimens. Furthermore, severe thermal damage was present in arteries, veins, and nerves. CONCLUSIONS: Intraoperative coating with fibrin glue can serve as a shield to protect vital structures such as arteries, veins, and nerves from accidental CO2 laser exposure. PMID- 9738765 TI - Scar recollagenation. AB - OBJECTIVES: This study was designed to show that native collagen could be regenerated in an area of prior tissue loss. STUDY DESIGN: Prospective. METHODS: Preserved, irradiated human cadaver fascia lata was inserted intradermally to restore topographic skin irregularities. Graft insertion was performed through a skin perforation created by a sterile, hypodermic 20-gauge needle hole into an intradermal pocket. Eighteen subjects with posttraumatic depressions were treated over a 13-month period and examined periodically afterward. Biopsies of selected sites were taken. RESULTS: Of the 44 defects treated in the study group, 14 were judged to be excellent or resolved, 23 were improved, and seven were unchanged at 24 weeks' follow-up. Biopsies showed graft preservation and fibroblast invasion at 12 weeks. DISCUSSION: Recollagenation is the application of a stimulator material into an area of prior collagen destruction to induce collagen production. Commonly, banked human cadaver fascia has been used to enhance the tissue volume in depressed acne scars, but this process has been successfully performed on a variety of postinflammatory and posttraumatic lesions. The process of fascia graft assimilation into human skin is demonstrated histologically. CONCLUSION: Recollagenation is an effective process for elevation of depressed, posttraumatic scars. PMID- 9738766 TI - Diagnosis, management, and follow-up of congenital head and neck teratomas. AB - OBJECTIVE: To describe our clinical experience with congenital teratomas of the head and neck. STUDY DESIGN: A retrospective review of the six patients as well as a review of the literature in the setting of academic referral centers. METHODS: In six infants with teratomas, four in the cervical region and two arising from the nasopharynx, surgical excision of all tumors was performed. Outcome measures were clinical and radiographic follow-up and the use of a fetoprotein (AFP) for postoperative monitoring. RESULTS: There was no recurrence of teratomas. CONCLUSIONS: Surgical excision is the treatment for congenital teratomas. Postoperative monitoring for recurrences should include AFP levels in difficult cases. PMID- 9738767 TI - Management of thyroid carcinoma invading the aerodigestive tract. AB - OBJECTIVES: To evaluate approaches to thyroid carcinoma invading the aerodigestive tract, with particular attention to well-differentiated carcinomas. STUDY DESIGN: Retrospective review of experience with thyroid carcinoma invading the aerodigestive tract over a 20-year period at a tertiary referral hospital. METHODS: The medical records of all patients with a diagnosis of thyroid cancer treated at Emory University Hospital, Atlanta, Georgia, from 1977 through 1997 were reviewed. Multiple clinical variables were analyzed including treatment, development of recurrence, and survival. Survival and time to local recurrence were determined by Kaplan-Meier analysis, and statistical comparisons were made using log-rank analysis. RESULTS: Five hundred thirty-six cases were identified; 28 patients (5.2%) were identified with invasive disease involving the aerodigestive tract. Histologic findings at the time of invasion included 15 well differentiated (WD) carcinomas and 13 poorly differentiated (PD) carcinomas. Eight of the 28 patients (5 WD, 3 PD) underwent surgical resection of some portion of the aerodigestive tract with curative intent. Ten patients (8 WD, 2 PD) underwent incomplete resection with tumor left adjacent to aerodigestive tract structures. All patients undergoing incomplete resection developed local recurrence. Six required salvage resection, as opposed to no recurrences in WD carcinomas following complete resection (P = .01). Survival at 5 years for WD carcinomas undergoing complete resection versus initial incomplete resection was 100% versus 50%, respectively (P = .27). CONCLUSION: Review of our experience shows that complete resection of thyroid carcinoma invading the aerodigestive tract can offer prolonged palliation, improved local control, and the opportunity for cure in selected patients. PMID- 9738768 TI - Middle fossa transpetrosal approach for petroclival and brainstem tumors. AB - OBJECTIVE: The purpose of the study was to demonstrate the utility of the middle fossa transpetrosal approach with anterior petrosectomy for difficult-to-access petroclival and pontine lesions. STUDY DESIGN: Retrospective case review in academic tertiary referral center. METHODS: Patients for inclusion had pontine and prepontine lesions of the petroclival region. Middle fossa transpetrosal approach with anterior petrosectomy with excision or biopsy of the lesion was performed. The main outcome measure was postoperative neurologic status including motor and cranial nerve function. RESULTS: No patient experienced neuromuscular compromise or cranial nerve deficits as a direct result of the surgical procedure. Complications consisted of a subdural temporal lobe hemorrhage and one case of cerebrospinal fluid rhinorrhea. CONCLUSIONS: The middle fossa transpetrosal approach with anterior petrosectomy was utilized for five patients with petroclival or pontine tumors. In this small series, it served well to spare cranial nerves and allowed avoidance of serious vascular injury. To our knowledge, this is the first reported use of this procedure for pontine venous angiomas. PMID- 9738769 TI - Overexpression of scatter factor and its receptor (c-met) in oral squamous cell carcinoma. AB - HYPOTHESIS: Scatter factor (SF) is a pleiotropic growth factor that recently has been shown to induce epithelial cell proliferation, random motility, and invasion via interaction with its receptor, a tyrosine kinase encoded by the c-met proto oncogene. Studies involving a variety of solid tumors have suggested that overexpression of the SF/c-met ligand-receptor pair is associated with the acquisition of a malignant phenotype. We hypothesize that SF and c-met are overexpressed in epithelial malignancies of the head and neck including squamous cell carcinoma (SCC) of the oral cavity. STUDY DESIGN: Immunohistochemical staining of randomly selected normal, dysplastic, and malignant oral tissues. METHODS: Formalin-fixed, paraffin-embedded tissues were obtained from the Department of Oral Pathology at Shands Hospital (University of Florida), Gainesville, Florida. Examples of mild dysplasia, severe dysplasia, well differentiated SCC, moderately differentiated SCC, and poorly differentiated SCC were randomly selected from the dictated reports of one of two staff oral pathologists. Histologically normal margins of each specimen served as normal controls. The tissues were immunohistochemically stained using commercially available antibodies against SF and c-met. Appropriate negative controls were run with each batch to ensure staining specificity. Evaluation of staining intensity was carried out using a computerized image analysis system. A one-way analysis of variance (ANOVA) with pairwise multiple-comparison procedures (Fisher method) was used to analyze the data. RESULTS: Statistically significant differences (P < .0001) in the intensity of staining were noted between the malignant and normal and the malignant and dysplastic tissues for both SF and c-met. No differences were appreciated when staining of normal and dysplastic sections of the SF stained tissue were compared. CONCLUSIONS: The results suggest that the SF/c-met ligand-receptor pair is overexpressed in SCC of the oral cavity. PMID- 9738770 TI - Clinical evoked electromyography for recurrent laryngeal nerve preservation: use of an endotracheal tube electrode and a postcricoid surface electrode. PMID- 9738771 TI - Z-meatoplasty of the external auditory canal. PMID- 9738772 TI - Toxin-mediated streptococcal and staphylococcal disease. AB - After several decades of seemingly decreasing virulence, streptococcal and staphylococcal infections have reemerged as a major source of morbidity and mortality. Within the past 2 decades, not only have well-established diseases such as rheumatic fever begun to reappear. but also many new entities, such as toxic shock syndrome, streptococcal toxic shock syndrome, recurrent toxin mediated perineal erythema, and recalcitrant erythematous desquamating disorder have been described. Central to the renewed importance of these bacteria has been the production of circulating toxins, which often function as superantigens in causing the clinical manifestations, morbidity and mortality associated with these diseases. PMID- 9738773 TI - Cutaneous manifestations of Paecilomyces lilacinus infection induced by a contaminated skin lotion in patients who are severely immunosuppressed. AB - BACKGROUND: New opportunistic fungal infections cause significant morbidity and death in patients who are severely immunocompromised. Cutaneous lesions may be the first clinical manifestation and give the clue to early diagnosis. OBJECTIVE: The purpose of this study was to describe the clinical and histologic manifestations of Paecilomyces lilacinus infection in patients who are severely immunosuppressed. METHODS: Within a 3-month period, we observed 5 patients with allogenic bone marrow transplantation and 4 patients with aplasia after chemotherapy for hematologic malignancies who developed skin eruptions caused by invasive P lilacinus. RESULTS: The skin lesions began in 7 cases during or shortly after recovery of pancytopenia. Most of the skin lesions were located on the lower extremities. The cutaneous manifestations were highly variable including erythematous macules, nodules, pustules, vesicular lesions, and necrotic crusts. In 3 biopsy specimens, histologic examination revealed hyphae in periodic acid-Schiff-stained sections. In all patients P lilacinus was isolated from skin tissue samples. P lilacinus was identified from all lesions either by skin biopsy or needle aspiration from clinically evident lesions. In 3 additional cases, the patient's hands were colonized without skin lesions. The source of the epidemic outbreak was finally traced down to several contaminated lots of a topical moisturizing agent. Two patients died; one patient had septic lesions in the eye and kidney as the result of P lilacinus. CONCLUSION: Clinical and histologic findings of P lilacinus infection with cutaneous manifestations in patients who are severely immunosuppressed are summarized. P lilacinus is resistant to all systemic antimycotics available, and in general, recovery of immunosuppression is necessary to eradicate the mold. Contaminated topical dermatologic agents should be included in the differential diagnosis as a source for severe epidemic cutaneous manifestations of fungal infection in patients who are severely immunosuppressed. This fact implies that additional safety guidelines are necessary for topical dermatologic agents used for patients who are severely immunosuppressed. PMID- 9738774 TI - Onychomycosis associated with Onychocola canadensis: ten case reports and a review of the literature. AB - BACKGROUND: Onychocola canadensis is a nondermatophyte mold associated with onychomycosis particularly in temperate climates (eg, Canada, New Zealand, and France). The slow growth rate of O canadensis and lack of resemblance to any other known nail-infecting fungus may have delayed its discovery. We are aware of 23 mycologically confirmed cases of O canadensis in the literature. OBJECTIVE: We describe 10 previously unreported Canadian patients, specimens from whom grew O canadensis. We also review the literature on infections associated with this organism. METHODS: Cases of O canadensis onychomycosis were diagnosed on the basis of (1) the finding of compatible filaments on direct microscopy of nail and (2) consistent culture from repeated specimens. All patients from whom O canadensis was isolated were followed up, but those in whom outgrowth was not consistent were not accepted as having "authentic" infections. RESULTS: In 10 patients O canadensis was found to be associated with distal lateral subungual onychomycosis (6 patients), white superficial onychomycosis (1 patient), and as an insignificant contaminant in the nails of 3 patients. Less commonly the organism may cause tinea manuum or tinea pedis interdigitalis. O canadensis appears to be more frequent in the elderly, especially females. It is not unusual for a patient with onychomycosis caused by O canadensis to be a gardener or farmer, suggesting that the infectious inoculum may originate from the soil. The optimal therapy for onychomycosis caused by this organism remains unclear. CONCLUSION: O canadensis may be the etiologic agent of distal and lateral subungual or white superficial onychomycosis; however, it may sometimes be present in an abnormal-appearing nail as an insignificant finding, not acting as a pathogen. PMID- 9738775 TI - Epidermolytic palmoplantar keratoderma with woolly hair and dilated cardiomyopathy. AB - BACKGROUND: A new cardiocutaneous syndrome has been noted, characterized by an epidermolytic palmoplantar keratoderma and woolly hair, and associated with dilated cardiomyopathy. OBJECTIVE: This describes the clinical and histopathologic characteristics of this new syndrome. METHODS: Eighteen patients were examined clinically and histologically from 1970 to 1997. Cardiologic examinations were performed in 12 patients. The cutaneous lesions were classified according to the presence of obligatory and facultative elements of the syndrome. RESULTS: Patients were born with woolly hair. Around the first year palmoplantar keratoderma and the other keratotic elements appeared. The first cardiac abnormalities are exclusively electrocardiographic and occur in asymptomatic patients. In these patients, dilation of the left ventricle together with alterations in muscle contractility are observed. The dilated cardiomyopathy can lead to congestive heart failure and death. CONCLUSION: The association of woolly hair and palmoplantar keratoderma with a histopathologic pattern of epidermolytic hyperkeratosis has not been previously described. Their frequent association with dilated cardiomyopathy defines a cardiocutaneous syndrome. Whenever woolly hair is associated with any kind of palmoplantar keratoderma, a search for possible cardiac abnormalities is recommended. PMID- 9738776 TI - Multiple primary melanomas. AB - BACKGROUND: The diagnosis of primary melanoma increases the risk of additional primary melanomas. OBJECTIVE: We characterize the subgroup of patients with multiple melanomas. METHODS: We reviewed the melanoma database. RESULTS: Sixty patients with multiple primary melanomas were identified. Twelve (20%) experienced melanomas in the same regional location, 43 (72%) in different locations, and 5 (8%) in both the same and different locations (> 2 melanomas). Eighteen (30%) were diagnosed concurrently with multiple melanomas, 38 (63%) subsequently, and 4 (7%) concurrently and subsequently (>2 melanomas). Forty-two percent of subsequent melanomas occurred within 3 years of the initial lesion diagnosis, 9 (17%) between 3 and 7 years, and 22 (42%) after more than 7 years. Subsequent melanomas were thinner in 70% of cases (P = .05). The mean age at first melanoma diagnosis was 38 and 59 years, respectively, for those with and without dysplastic nevi (P < .001). CONCLUSION: In patients with multiple melanomas, subsequent melanomas often occur in different regional locations several years after diagnosis of the initial lesion. PMID- 9738778 TI - Antinuclear antibody seropositivity in patients with cutaneous T-cell lymphoma. AB - BACKGROUND: We attempted to determine the frequency and clinical relevance of antinuclear antibody (ANA) testing and positive ANA test results in patients with cutaneous T-cell lymphoma (CTCL). METHODS: A retrospective chart and computer record review was conducted to determine the frequency of ANA testing in CTCL patients and the rate of seropositivity. Patients with a positive ANA were further examined to define possible explanations of the positive test. RESULTS: Of 381 patients with CTCL, 66 (17%) had ANA tests; 8 of these (12.1%) were found to have an ANA titer greater than or equal to 1:40. Of patients with a positive ANA test, one was found to have chronic cutaneous lupus erythematosus histologically and clinically mimicking CTCL. Others were found to have a comorbid connective tissue disorder, some had apparent drug-induced antinuclear antibodies, and some had no identifiable reason for a positive ANA test. CONCLUSION: ANA seropositivity does not appear to be increased in CTCL patients, and the ANA test remains a useful screening tool for differentiating between CTCL and connective tissue disorders. PMID- 9738777 TI - Risk of melanoma in medium-sized congenital melanocytic nevi: a follow-up study. AB - BACKGROUND: The risk of the occurrence of malignant melanoma (MM) in medium-sized (1.5 to 19.9 cm in diameter) congenital melanocytic nevi (CMN) is the subject of controversy. Universally accepted recommendations regarding the management of such lesions have not been made. OBJECTIVE: Our purpose was to assess the risk of MM arising in medium-sized CMN. METHODS: The study included 230 medium-sized CMN in 227 patients, first seen in a private dermatology practice from 1955 to 1996, who were followed up for MM arising within their CMNs. Criteria for entry into the study included (1) a clinically diagnosed medium-sized CMN, (2) minimum follow-up period of 1 year, and (3) a photograph of the lesion in the patient's medical record. RESULTS: No MM occurred in a medium-sized CMN during an average follow-up of 6.7 years (median, 5.8 years) to an average age of 25.5 years (median, 19.1 years). CONCLUSION: The results of this short-term follow-up study do not support the view that there is a clinically significantly increased risk for MM arising in banal-appearing medium-sized CMN or that prophylactic excision of all such lesions is mandatory. Lifelong medical observation seems a reasonable alternative for many medium-sized CMN. PMID- 9738779 TI - Diffuse plane xanthoma: clinicopathologic study of 8 cases. AB - BACKGROUND: Diffuse plane xanthoma (DPX) usually has been described in association with a reticuloendothelial disease. However, the incidence of an underlying disease and its relation to clinical and histopathologic features have not been previously described. OBJECTIVE: We describe the clinicopathologic features of a series of patients with DPX. METHODS: Eight patients with DPX were diagnosed and studied between 1987 and 1996. RESULTS: Three of the 8 patients had a reticuloendothelial disease (benign monoclonal gammopathy in 2 and chronic myelomonocytic leukemia in 1). These 3 patients had larger cutaneous lesions and involvement of trunk and extremities. Histologic examination showed only foamy macrophages in 2 of the 8 patients. The remainder exhibited macrophages, foamy histiocytes, lymphocytes, and Touton cells. CONCLUSION: The incidence of underlying disease associated with DPX seems to be lower than expected. However, patients with DPX should be observed for the potential occurrence of an associated condition. PMID- 9738780 TI - Kinetics of photosensitivity in bath-PUVA photochemotherapy. AB - BACKGROUND: Bath-PUVA is used to treat a variety of dermatoses. However, the kinetics of 8-methoxypsoralen during treatment are not completely clarified. OBJECTIVE: The purpose of this study was to investigate the intensity of the phototoxic response and the persistence of phototoxicity after bath-PUVA. METHODS: Twelve volunteers were exposed to UVA doses ranging from 0.5 to 40 J/cm2 from 10 to 240 minutes after bath-PUVA treatment. The resulting phototoxic response of the skin was determined. RESULTS: Irradiation 10 minutes after the psoralen bath led to the lowest assessed minimal phototoxic dose (MPD) of 1.42 J/cm2 (mean, SD +/- 0.29). Thereafter, the MPD increased significantly and sharply every hour. At 4 hours after the psoralen bath, UVA doses up to 40 J/cm2 failed to induce any phototoxic erythema (MPD). CONCLUSION: For optimal effects, UVA irradiation has to be administered immediately after the psoralen bath; no restrictive behavior is necessary after bath-PUVA treatment. PMID- 9738781 TI - Calcipotriene ointment and halobetasol ointment in the long-term treatment of psoriasis: effects on the duration of improvement. AB - BACKGROUND: Weekend therapy with superpotent topical corticosteroids has been used for the long-term treatment of psoriasis. Recently, calcipotriene ointment has been added to this regimen for use on weekdays, but there are no long-term studies of that combination. OBJECTIVE: The purpose of this study was to determine whether the addition of weekday calcipotriene to a pulse therapy regimen of weekend superpotent corticosteroids results in a longer duration of remission of plaque psoriasis. SUBJECTS: This was a double-blind, placebo controlled, parallel-group study. Forty-four patients with mild to moderate psoriasis were treated with calcipotriene ointment in the morning and halobetasol ointment in the evening for 2 weeks. Thereafter, 40 patients who were at least moderately (50% or greater) improved were randomized to 2 treatment groups. After 2 weeks of treatment with calcipotriene ointment in the morning and halobetasol ointment in the evening, 20 patients were randomized to receive halobetasol ointment twice daily on weekends and calcipotriene ointment twice daily on weekdays, and 20 patients were randomized to receive halobetasol ointment twice daily on weekends and placebo ointment twice daily on weekdays. RESULTS: Seventy six percent of patients applying halobetasol ointments on weekends and calcipotriene ointment on weekdays were able to maintain remission for 6 months compared with 40% of patients applying halobetasol ointment on weekends only with the vehicle on weekdays. CONCLUSION: The addition of calcipotriene ointment applied on weekdays to a weekend pulse therapy regimen of superpotent corticosteroids can increase the duration of remission of psoriasis. PMID- 9738782 TI - Laboratory tests and imaging studies in patients with cutaneous malignant melanoma. AB - Laboratory tests and imaging studies are often ordered for asymptomatic patients with malignant melanomas (MMs) in an effort to detect subclinical metastatic disease. However, their sensitivity and specificity for detecting cryptic metastases are not well established. A review of the literature on laboratory tests and imaging studies for MM metastases was undertaken to address the usefulness of such investigations in asymptomatic patients with MM in AJCC (American Joint Committee on Cancer system of classification) stages I, II, and III. A review of the pertinent literature since 1966 was conducted through MEDLINE, Medica, and Cancerlit. Laboratory tests and imaging studies revealed occult MM metastases in only a small number of the thousands of reported patients with putative AJCC stage I, II, and III MM. However, for those diagnosed with limited metastases, surgical removal with or without immunotherapy, chemotherapy, or radiotherapy can lead to long-term remissions in some patients. For patients with asymptomatic AJCC stage I or II disease, chest roentgenograms (CXR) and blood lactic dehydrogenase (LDH) levels may be obtained at low cost and prove to be of benefit if metastases are identified. For patients with AJCC stage III disease, computed tomographic (CT) scans of the thorax, abdomen, and pelvis (especially when the primary cutaneous site of the melanoma is below the waist) may be considered for detecting metastatic MM. Other tests, such as magnetic resonance imaging (MRI) scans of the brain, may be ordered based on symptoms or physical findings. In the future, technologically improved techniques and newer methods may prove cost-effective for detecting treatable asymptomatic MM metastases. Furthermore, improvement in treatments will also influence the indications for the search for occult MM metastases. At this time there is a need for an international consensus conference on laboratory tests and imaging studies for occult melanoma metastases. PMID- 9738783 TI - Cyclosporine consensus conference: with emphasis on the treatment of psoriasis. AB - Cyclosporine has been in worldwide use for 15 years for patients who have undergone transplantation operations and is now being used to control inflammatory reactions in other organs (eg, joints, bowel, and skin). Neoral, a more consistently absorbed form of cyclosporine, has recently been approved by the Food and Drug Administration for the treatment of psoriasis. This report outlines the indications, contraindications, dosage recommendations, monitoring requirements, adverse events, drug interactions, interactions with other psoriasis treatments, and suggestions for cyclosporine's use in rotational therapy. PMID- 9738784 TI - Surgical pearl: Dermal advancement flaps for filling deep dorsal nasal defects under grafts. PMID- 9738785 TI - Useful plants of dermatology. IV. Alizarin red and madder. PMID- 9738786 TI - Reversible transverse overcurvature of the nails (pincer nails) after treatment with a beta-blocker. PMID- 9738787 TI - A reaction to a red lip cosmetic tattoo. PMID- 9738788 TI - Longitudinal melanonychia revealing an intraepidermal carcinoma of the nail apparatus: detection of integrated HPV-16 DNA. PMID- 9738789 TI - Isolation of Mycobacterium avium complex from erythema multiforme. PMID- 9738790 TI - Intertriginous granular parakeratosis. PMID- 9738791 TI - Successful therapy for bullous pemphigoid with mycophenolate mofetil. PMID- 9738793 TI - Bullous lichen sclerosus et atrophicus. PMID- 9738792 TI - Scedosporium apiospermum skin infection: a case report and review of the literature. PMID- 9738795 TI - A familial dermatofibrosarcoma protuberans. PMID- 9738794 TI - The efficacy of metronidazole 1% cream once daily compared with metronidazole 1% cream twice daily and their vehicles in rosacea: a double-blind clinical trial. PMID- 9738797 TI - Treatment of relapsing idiopathic nodular panniculitis (Pfeifer-Weber-Christian disease) with mycophenolate mofetil. PMID- 9738796 TI - Multiple myeloma in an HIV-positive man presenting with primary cutaneous plasmacytomas and spinal cord compression. PMID- 9738798 TI - Congenital leukonychia. PMID- 9738799 TI - Cutaneous Paecilomyces lilacinus infection. PMID- 9738800 TI - High-dose UVA1 radiation therapy for localized scleroderma. PMID- 9738801 TI - Antimicrobial effects of lidocaine, bicarbonate, and epinephrine. PMID- 9738802 TI - Usefulness of the staged excision for lentigo maligna and lentigo maligna melanoma. PMID- 9738803 TI - The National Cancer Data Base report on cancer of the head and neck. AB - BACKGROUND: The National Cancer Data Base (NCDB), a large sample of cancer cases accrued from hospital-based cancer registries, is sponsored by the Commission on Cancer of the American College of Surgeons and the American Cancer Society. The NCDB permits a detailed analysis of case-mix, treatment, and outcome variables. OBJECTIVE: To provide an overview of the contemporary status of the subset of patients with head and neck cancer in the United States. METHODS: The NCDB, which obtains data from US as well as Canadian and Puerto Rican hospitals, accrued 4 583 455 cases of cancer between 1985 and 1994. Of these cases, 301350 (6.6%) originated in the head and neck. We address 295022 cases of head and neck cancer limited to the 50 United States and District of Columbia. Cases were segregated into an earlier group (1985-1989) to permit 5-year follow-up and into a later group (1990-1994) to analyze a more contemporary group. Comparison between both periods permits identification of trends. RESULTS: The largest proportion of cases arose in the larynx (20.9%) and oral cavity, including lip (17.6%) and thyroid gland (15.8%). Squamous cell carcinoma (55.8%) was the most common histological finding, followed by adenocarcinoma (19.4%) and lymphoma (15.1%). Income level (low), race (African American), and tumor grade (poorly differentiated) were most notably associated with advanced stage. Treatment was most commonly surgery alone (32.4%), combined surgery with irradiation (25.0%), and irradiation alone (18.9%). Overall 5-year, disease-specific survival was 64.0%. Cancer of the lip demonstrated the best survival (91.1%) and cancer of the hypopharynx the worst survival (31.4%). CONCLUSIONS: This NCDB analysis of cancer of the head and neck provides a contemporary overview of head and neck cancer in the United States. It also serves to introduce a series of NCDB articles that address specific anatomical sites and histological types through separate, detailed analysis. PMID- 9738805 TI - Prognostic staging system for recurrent, persistent, and second primary cancers of the oral cavity and oropharynx. AB - OBJECTIVE: To develop a practical staging system for predicting mortality of patients with recurrent squamous cell tumors of the oral cavity and oropharyngeal mucosa. DESIGN AND SETTING: An inception cohort at an academic medical center. PATIENTS: A total of 308 patients who had evidence of recurrent, persistent, or second primary tumors of the oral cavity and oropharynx between January 1, 1980, and December 31, 1991, of whom 162 (52.6%) met inclusion criteria. MAIN OUTCOME MEASURE: One-year mortality. RESULTS: The median survival time was 10 months. In bivariate analysis, the TNM stage of the recurrent tumor, invasion of pharyngeal constrictors and the floor-of-mouth muscles, weight loss, local and systemic symptoms, and eating function had significant effects on mortality. Multivariable analysis (done by conjunctive consolidation and Cox regression) identified constrictor invasion, the TNM stage of the recurrence, and weight loss as having a substantial effect on mortality. A composite 4-stage system using these 3 variables demarcated 1-year survival rates of 88.2% (30/34), 71.9% (23/32), 32.6% (16/49), and 4.2% (2/47). CONCLUSIONS: The TNM status of recurrent tumors predicts mortality, but constrictor muscle invasion and weight loss also have major prognostic importance. The consolidation of these variables into a composite staging system successfully stratifies patients with widely divergent mortality rates. Improved staging of recurrent head and neck tumors can lead to more effective decisions about the comparisons and merits of additional treatment. PMID- 9738804 TI - Long-term quality of life after treatment of laryngeal cancer. The Veterans Affairs Laryngeal Cancer Study Group. AB - OBJECTIVE: To assess long-term quality of life in surviving patients with advanced laryngeal cancer. DESIGN: A follow-up long-term quality-of-life survey of patients randomized to the Veterans Affairs Laryngeal Cancer Study No. 268 on induction chemotherapy and radiation (CT + RT) vs surgery and RT. SETTING AND PATIENTS: Forty-six (71%) of the 65 surviving patients with prior stage III or IV laryngeal cancer who could be contacted completed the survey: 25 from the surgery and RT group and 21 from the CT + RT group. Baseline demographic and clinical characteristics among survey respondents were similar, except that those in the CT + RT group were significantly older (mean, 61.2 years) than those in the surgery and RT group (mean, 55.7 years; P<.05). INTERVENTIONS AND MAIN OUTCOME MEASURES: Patients completed the University of Michigan Head and Neck Quality of Life (HNQOL) instrument, the Medical Outcomes Studies Short-Form 36 (SF-36) general health survey, the Beck Depression Inventory as well as smoking and alcohol consumption surveys. RESULTS: Patients randomized to the CT + RT group had significantly better (P<.05) quality-of-life scores on the SF-36 mental health domain (76.0) than the surgery and RT group (63.0), and also had better HNQOL pain scores (81.3 vs 64.3). Compared with patients who underwent laryngectomy, patients with intact larynges (CT + RT with larynx) had significantly less bodily pain (88.5 vs 56.5), better scores on the SF-36 mental health (79.8 vs 64.7), and better HNQOL emotion (89.7 vs 79.4) scores. More patients in the surgery and RT group (28%) were depressed than in the CT + RT group (15%). CONCLUSION: Better quality-of-life scores in the CT + RT groups appear to be related to more freedom from pain, better emotional well-being, and lower levels of depression than to preservation of speech function. PMID- 9738806 TI - Osteoinduction using bone morphogenic protein in irradiated tissue. AB - OBJECTIVE: To prove the efficacy of bone morphogenic protein as an osteoinductive agent in irradiated tissue. DESIGN: Prospective randomized controlled trial designed to test the effectiveness of recombinant bone morphogenic protein 2 (rBMP-2) combined with solid hydroxyapatite disks in an irradiated tissue bed. SUBJECTS: Eighteen adult, male, white New Zealand rabbits weighing 3.0 to 3.5 kg. INTERVENTION: The rabbits were randomly divided, with 9 receiving radiation treatment and 9 receiving no radiation treatment. Each animal underwent implantation of 2 hydroxyapatite disks onto the snout at 9 weeks following radiation treatment. One disk was impregnated with rBMP-2 and the other with buffer only. The animals were killed at 3, 6, or 20 weeks after implantation for analysis. RESULTS: Histological analysis demonstrated that rBMP-2 was equally effective as an osteoinductive agent in the irradiated and nonirradiated tissue. We also found significantly increased new bone formation in the rBMP-2 group vs the buffer group. CONCLUSIONS: This study supports the potential clinical utility of rBMP-2 and solid hydroxyapatite in irradiated tissue beds. These findings have interesting implications for patients with head and neck cancer who have undergone radiation therapy and need bony reconstruction. PMID- 9738807 TI - How accurate is parent rating of hearing for children with otitis media? AB - OBJECTIVE: To determine the accuracy of parent assessment of child hearing. DESIGN: Prospective study. SETTING: Hospital-based pediatric otolaryngology practice in a metropolitan area. PATIENTS: One hundred eighty-six children aged 6 months to 12 years (median age, 3.4 years) with chronic otitis media with effusion or recurrent acute otitis media enrolled in a quality-of-life study. INTERVENTION: Parents rated their child's hearing over the prior 4 weeks using a 7-point response scale. Otoscopic findings, static admittance, tympanometric width, and audiometric thresholds were recorded concurrently. Fifty children were reassessed to monitor changes in hearing. MAIN OUTCOME MEASURE: Correlation of parent hearing assessments with baseline hearing status (pure tone average for the better hearing ear) and with changes in hearing status. RESULTS: The hearing loss questions had good test-retest reliability (R=0.79) but did not correlate with audiometric results (R=-0.13; P=.09). Only when caregivers reported hearing to be an "extreme problem" were median hearing levels (31 dB) significantly greater than the median response (20 dB). Conversely, static admittance and tympanometric gradient were significant predictors of hearing levels (2-way analysis of variance, P<.01) and explained 44% of the ear-specific variations. Abnormal immittance measures in both ears had an 84% predictive value for hearing loss (20-dB hearing level or poorer), and normal immittance measures in both ears had a 76% predictive value for normal hearing. Caregiver assessments of change in hearing status did not correlate with changes in audiometric results (R=0.07; P=.65). CONCLUSIONS: Caregiver assessments of child hearing do not accurately predict hearing levels or changes in hearing status. Immittance measures can help identify children at low or high risk for hearing loss, but cannot substitute for audiometry. PMID- 9738808 TI - Persistence of group A beta-hemolytic streptococci in toothbrushes and removable orthodontic appliances following treatment of pharyngotonsillitis. AB - OBJECTIVE: To investigate the persistence of group A beta-hemolytic streptococci (GABHS) in toothbrushes and removable orthodontic appliances (ROAs) in children who suffer from acute GABHS pharyngotonsillitis and the association with penicillin treatment failure. SETTING: Private practice setting. PATIENTS AND METHODS: Pharyngotonsillar and toothbrush cultures were obtained from 104 children with acute GABHS pharyngotonsillitis before and after 10 days of penicillin V potassium therapy. Cultures of ROAs were also obtained from 21 children. The persistence of GABHS in 10 daily rinsed and 10 nonrinsed toothbrushes was studied in vitro. RESULTS: Group A beta-hemolytic streptococci were isolated from 11 (11%) of the toothbrushes and 18 (17%) of the patients after the completion of penicillin therapy. Toothbrushes of 5 (28%) of the 18 children who harbored GABHS were colonized with the organism. Group A beta hemolytic streptococci were also isolated from 4 (19%) of 21 ROAs after therapy. In vitro studies illustrated the persistence of GABHS in nonrinsed toothbrushes for up to 15 days. In contrast, the organism was not isolated from rinsed toothbrushes beyond day 3. CONCLUSION: Toothbrushes and ROAs that harbor GABHS may contribute to the persistence of GABHS in the oropharynx and may account for the failure of penicillin therapy in some cases of pharyngotonsillitis. PMID- 9738809 TI - Evaluation of patients with sleep apnea after tracheotomy. AB - OBJECTIVE: To determine the effect of tracheotomy on polysomnographic and arterial blood gas data in patients with obstructive sleep apnea (OSA). DESIGN: A retrospective study of all patients who underwent tracheotomy and were studied polysomnographically at the Johns Hopkins Sleep Disorders Center, Baltimore, Md, since 1981. SETTING: A regional sleep disorders center. PATIENTS: Twenty-eight patients (8 women and 20 men), aged 22 through 77 years. Patients were categorized into 2 groups on the basis of whether they had already undergone tracheotomy before polysomnography. Group 1 patients all had a polysomnographic diagnosis of OSA before tracheotomy. They were further subdivided on the basis of whether cardiopulmonary decompensation had been absent (group 1a, n=10) or present (group 1b, n=13). Group 2 patients (n=5) had undergone tracheotomy to treat upper airway obstruction that developed after non-apnea-related upper aerodigestive tract surgeries. INTERVENTION: Tracheotomy. MAIN OUTCOME MEASURES: Nocturnal non-rapid eye movement, apnea-hypopnea index, percentage oxyhemoglobin saturation, and arterial blood gas data. RESULTS: Patients with OSA underwent tracheotomy as definitive treatment for the apnea (n=15), to prevent postoperative upper airway compromise after uvulopalatopharyngoplasty (n=7), and to treat upper airway compromise after non-apnea-related upper aerodigestive tract surgeries (n=6). Tracheotomy alleviated apnea in all 10 patients with uncomplicated sleep apnea (group 1a). For patients with OSA complicated by cardiopulmonary decompensation (group 1b), tracheotomy improved but did not eliminate sleep apnea in 7 of the 13 patients, despite overall improvement in arterial blood gas values. For patients whose sleep apnea had not been diagnosed polysomnographically before tracheotomy (group 2), tracheotomy was still required to treat OSA that had previously not been recognized. CONCLUSIONS: Tracheotomy effectively treated patients with uncomplicated OSA, but was much less effective in treating patients with OSA and cardiopulmonary decompensation. In patients who underwent tracheotomy in conjunction with other upper aerodigestive tract surgeries, concomitant obstructive sleep apnea often required continued use of a tracheotomy to maintain upper airway patency. PMID- 9738810 TI - Efficacy of fiberoptic laryngoscopy in the diagnosis of inhalation injuries. AB - BACKGROUND: A significant proportion of burn patients with inhalation injuries incur difficulties with airway protection, dysphagia, and aspiration. In assessing the need for intubation in burn patients, the efficacy of fiberoptic laryngoscopy was compared with clinical findings and the findings of diagnostic tests, such as arterial blood gas analysis, measurement of carboxyhemoglobin levels, pulmonary function tests, and radiography of the lateral aspect of the neck. OBJECTIVE: To determine if these patients were at risk for aspiration or dysphagia, barium-enhanced fluoroscopic swallowing studies were performed. DESIGN: Prospective study. SETTINGS: Burn intensive care unit in an academic tertiary referral center. MAIN OUTCOME MEASURES: Need for endotracheal intubation and potential for aspiration. RESULTS: Six (55%) of 11 patients had clinical findings and symptoms that indicated, under traditional criteria, endotracheal intubation for airway protection. Visualization of the upper airway with fiberoptic laryngoscopy obviated the need for endotracheal intubation in all 11 patients. These patients also failed to evidence an increased risk of aspiration or other swallowing dysfunction. CONCLUSIONS: In comparison with other diagnostic criteria, fiberoptic laryngoscopy allows differentiation of those patients with inhalation injuries who, while at risk for upper airway obstruction, do not require intubation. These patients may be safely observed in a monitored setting with serial fiberoptic examinations, thus avoiding the possible complications associated with intubation of an airway with a compromised mucosalized surface. In these patients, swallowing abnormalities do not manifest. PMID- 9738811 TI - Erythema after cutaneous laser resurfacing using a porcine model. AB - OBJECTIVES: To measure and compare postoperative erythema after laser cutaneous resurfacing using 2 carbon dioxide laser systems and varying postoperative treatment methods. DESIGN: Carbon dioxide laser systems are used as cutaneous resurfacing tools. The continuous-wave lasers have been associated with postoperative erythema, but the short-pulsed lasers reportedly result in less postoperative erythema because of shorter pulse durations. Although subjective evaluations of results have been published, a side-by-side comparison with digital photography has not been performed. Furthermore, postoperative treatment varies among physicians, and objective data about this treatment are scarce. SUBJECTS: To compare postoperative erythema, we created 240 resurfacing wounds on 8 piglets with continuous-wave and short-pulsed lasers, using the manufacturers' suggested settings. By using photography and computed color analysis, we measured the resultant erythema after 1, 3, and 5 laser passes at days 0, 1, 3, 5, 7, and 14. Tissue samples were obtained for histological analysis on days 0, 3, 7, and 14. INTERVENTION: We compared the resolution of erythema after postoperative treatment with petroleum jelly (Vaseline), a wound dressing (Vigilon), partially hydrogenated vegetable oil (Crisco), or a combination drug, bacitracin zinc neomycin sulfate-polymyxin B sulfate on the wounds. RESULTS: The short-pulsed carbon dioxide laser resulted in an average of 22% less erythema compared with the continuous-wave laser (P<.001). No statistically significant difference in erythema was found among the postoperative treatment methods (P>.10). CONCLUSIONS: Compared with the continuous-wave laser, the short-pulsed carbon dioxide laser results in less postoperative erythema. However, the type of postoperative treatment has little, if any, beneficial effect for reducing erythema. PMID- 9738812 TI - Report of the first case of invasive fungal sinusitis caused by Scopulariopsis acremonium: review of scopulariopsis infections. AB - Scopulariopsis acremonium is a species of saprophytic fungus not previously reported to cause invasive disease in humans, although invasive infections from other species of Scopulariopsis have been reported and are reviewed. Deep infection with this fungus is associated with a high mortality rate. Invasive fungal sinusitis, in general, is a potentially fatal disease that typically affects immunocompromised patients, such as those receiving intensive chemotherapy or undergoing bone marrow transplantation. We report a case of invasive fungal sinusitis caused by Scopulariopsis acremonium in a patient with leukemia, who was successfully treated with amphotericin B, itraconazole, endoscopic sinus surgery, and granulocyte colony-stimulating factor. PMID- 9738813 TI - Self-inserted sphenoid sinus foreign bodies. AB - This report describes the case of a patient who inserted 3 foreign bodies into her right sphenoid sinus. The possible consequences of sphenoid sinus foreign bodies are described, as well as the importance of a psychiatric workup in such cases. PMID- 9738814 TI - Endoscopic cauterization for treatment of fourth branchial cleft sinuses. AB - Fourth branchial cleft sinuses are rare, and the nature of their origin is controversial. Clinical presentation is varied because they may present as asymptomatic neck masses, recurrent neck abscesses, or suppurative thyroiditis. We describe herein 7 children who presented with abscesses on the left side of their necks, 3 of whom had abscesses that involved the thyroid gland. Direct laryngoscopy revealed that all 7 children had a sinus tract opening into the apex of the piriform sinus. Endoscopic obliteration of this tract was achieved using an insulated electrocautery probe either when the abscess was initially incised and drained or 4 to 6 weeks later. All 7 children recovered uneventfully. Four of the 7 children were followed up for more than 18 months without recurrence. PMID- 9738815 TI - Subclavian pseudoaneurysm mimicking recurrent head and neck cancer. AB - Vascular blowouts are devastating complications of head and neck oncologic surgery that are easily diagnosed with oropharyngeal or external bleeding. We present herein a case of a pseudoaneurysm of the subclavian artery that mimicked recurrent head and neck carcinoma. Vascular lesions arising in the base of the neck may present with few signs of vascular injury. Head and neck surgeons should be aware of this unusual complication to avoid a potentially life-threatening event. We report difficulties in diagnosing subclavian pseudoaneurysm, a review of the vascular injuries related to this condition, as well as approaches to bleeding at the base of the neck. PMID- 9738816 TI - Laryngeal glycogenic acanthosis presenting as leukoplakia. AB - We report a case of an incidental finding of glycogenic acanthosis of the larynx on autopsy in a 79-year-old man who died of myocardial infarction. The lesion was grossly recognized as a white plaque (leukoplakia) on the subglottic compartment of the left side of the larynx. Histological sections revealed thickened squamous mucosa positive for abundant glycogen on periodic acid-Schiff staining. No epithelial dysplasia was noted. The patient had a history of smoking. This case represents the first report of glycogenic acanthosis involving the larynx. This benign condition should be added to the vast differential diagnosis of leukoplakia in this anatomical location. PMID- 9738817 TI - Synchronous glottic granular cell tumor and subglottic spindle cell carcinoma. AB - We report the clinicopathologic findings in a case of coexistent glottic granular cell tumor and subglottic spindle cell carcinoma. There is no evidence in this case that suggests malignant transition of the granular cell tumor to malignant tumor. To our knowledge, this is the first report of synchronous granular cell tumor and subglottic spindle cell carcinoma. PMID- 9738818 TI - Primary T-cell non-Hodgkin lymphoma of the larynx with subsequent cutaneous involvement. AB - BACKGROUND: T lymphocytes expressing the gammadelta T-cell receptor represent a minority of normal T lymphocytes and are mostly located in the spleen or mucosa. Lymphomas expressing the gammadelta T-cell receptor are rare and usually present as hepatosplenic (negative for Epstein-Barr virus) disease. Primary lymphomas of the larynx are also rare. OBJECTIVE: To report the first case of primary laryngeal gammadelta T-cell lymphoma related to Epstein-Barr virus infection. DESIGN: Single-case study, including clinical, histological, immunohistochemical, and ultrastructural analysis, and in situ hybridization for Epstein-Barr virus encoded small nuclear RNA. PATIENT: An 88-year-old man presenting with a 6-month history of a cough followed by progressive dysphonia and a thickening of the left aspect of the aryepiglottic fold. INTERVENTION: Two weeks of treatment with corticosteroids and antibiotics, followed by radiotherapy and then chemotherapy with chlorambucil and corticosteroids. OUTCOME: The patient died of heart failure 10 months after the onset of the disease. RESULTS: The tumor was laryngeal and disseminated to the skin over the parotid gland. Tumor cells were medium-sized T cells of cytotoxic immunophenotype, expressed the gammadelta T-cell receptor, and contained azurophilic granules and cytotoxiclike granules detected on electron microscopy. Epstein-Barr virus-encoded small nuclear RNA was detected in most tumor cells. CONCLUSIONS: Lymphomas with a T-cell cytotoxic phenotype expressing the gammadelta T-cell receptor are rare, and this case appears to be the first to involve the larynx. The association between Epstein-Barr virus and T-cell lymphomas has been shown to be frequent in the upper respiratory tract and is confirmed in this case. This finding suggests that T cells in the upper respiratory tract may be more exposed to Epstein-Barr virus infections, perhaps because of their anatomical location. PMID- 9738819 TI - Harold Hopkins. PMID- 9738820 TI - Is selective neck dissection really as efficacious as modified radical neck dissection for elective treatment of the clinically negative neck in patients with squamous cell carcinoma of the upper respiratory and digestive tracts? PMID- 9738821 TI - Selective neck dissection. PMID- 9738822 TI - Imaging quiz case 1. Mucocele. PMID- 9738823 TI - Imaging quiz case 2. Jugular bulb dehiscence. PMID- 9738824 TI - Unilateral parathyroid exploration. PMID- 9738825 TI - Unilateral vs bilateral parathyroid gland exploration: a continuing controversy. PMID- 9738826 TI - Surgical management of primary hyperparathyroidism: review of my experience at the University of Vermont. PMID- 9738827 TI - Antibiotic resistance. PMID- 9738828 TI - Extended-spectrum beta-lactamases in Pseudomonas aeruginosa. PMID- 9738829 TI - Interactions of plaunotol with bacterial membranes. AB - Plaunotol, a cytoprotective antiulcer agent, has a bactericidal effect against Helicobacter pylori, which may result from interaction of this compound with the bacterial cell membrane. The purpose of the present study was to confirm that plaunotol interacts with the H. pylori membrane. Membrane fluidities were measured using two stearic acid spin labels, namely 5-doxyl-stearic acid (in which the nitroxide group is located in the upper portion of the bacterial cell membrane) and 16-doxyl-stearic acid methyl ester (in which the nitroxide group is located deeper in the bacterial cell membrane), by means of electron spin resonance. The membrane fluidities of plaunotol-treated cells were significantly increased in the measurements made using the two spin labels. We also attempted to isolate plaunotol-resistant H. pylori in vitro by two different methods. To assess the level of resistance that could be reached, H. pylori was passaged five times on an agar plate containing subinhibitory concentrations of plaunotol or metronidazole. To measure the rate of development of resistance, H. pylori was grown with subinhibitory concentrations (0.25 x MIC) of plaunotol or metronidazole, and quantitatively plated on to medium containing 4 x MIC of the compounds. This treatment was repeated once more. No plaunotol-resistant colonies were selected by the two methods. H. pylori developed resistance to metronidazole easily and at a relatively high rate. The mechanism by which plaunotol directly fluidizes and destroys the H. pylori membrane might make it difficult for this organism to develop resistance to plaunotol. It was confirmed that the bactericidal effects of plaunotol were also shown against Staphylococcus aureus, Streptococcus pneumoniae, Neisseria gonorrhoeae, Moraxella catarrhalis and Haemophilus influenzae. No such effect was seen against Escherichia coli and Pseudomonas aeruginosa. PMID- 9738830 TI - In-vitro activity of lytic peptides, inhibitors of ion transport systems and ionophorous antibiotics against Pneumocystis carinii. AB - The in-vitro activity of two vertebrate lytic peptides, two ion transport system inhibitors and two polyether ionophores was investigated against four clinical isolates of Pneumocystis carinii recovered from bronchoalveolar lavages of AIDS patients. The susceptibility tests were performed by inoculating the isolates on to cell monolayers and determining the parasite count after 72 h incubation at 37 degrees C. The culture medium was supplemented with Dulbecco's modified Eagle's medium containing serial dilutions of cecropin P1, magainin II, benzamil, 5 (Nmethyl-Nisobutyl)amiloride (MIBA), lasalocid and nigericin. The two vertebrate lytic peptides showed high activity against trophozoites and cysts. At a concentration of 66.77 mg/L, cecropin P1 produced a 93.3% and 98.1% reduction in cyst and trophozoite counts, respectively, while magainin II at a concentration of 49.33 mg/L produced a 90.6% and 98.7% reduction in cyst and trophozoite counts, respectively. The IC50s of benzamil and MIBA were observed at the highest concentrations tested, 35.62 and 29.98 mg/L, respectively. However, 90% inhibition was not achieved. Lasalocid and nigericin at 0.05 mg/L gave inhibition comparable to that observed with the highest tested concentrations of cecropin P1 and magainin II, but significant injury to the cell monolayer was also observed when nigericin was tested at this concentration. Lasalocid 0.05 mg/L produced a reduction of 91.3% and 92.0% in cyst and trophozoite counts, respectively. Our results suggest that lytic peptides and lasalocid may be effective in inhibiting P. carinii growth at concentrations which are not toxic for the cell monolayer. PMID- 9738831 TI - Influence of ciprofloxacin and other antimicrobial drugs on different Escherichia coli strains in continuous-flow cultures under aerobic and anaerobic conditions. AB - In continuous-flow culture, long generation times and high bacterial counts favour survival of bacteria. A chemotherapeutic agent that achieves a bactericidal effect under these circumstances can therefore be seen as highly effective. In our continuous-flow culture we obtained bactericidal effects with ciprofloxacin 1-2 mg/L, cefotaxime 4 mg/L and mezlocillin 32 mg/L. These effects were seen irrespective of whether conditions were aerobic or anaerobic. There were no significant differences between monocultures and mixed cultures simulating faecal flora with the various Escherichia coli strains tested. Cefotaxime had an initial effect but an increase in counts was then observed as a result of regrowth of E. coli survivor strains in aerobic monoculture and mixed cultures. Mezlocillin was completely bactericidal in monocultures, but regrowth occurred in mixed cultures under anaerobic conditions. Neither the bacterial composition of this culture nor the resistance pattern explained this regrowth. These results were observed in long-term experiments followed for up to 7 days. We conclude that the antibiotics tested are highly effective against E. coli under unfavourable conditions simulating in-vivo situations. PMID- 9738832 TI - Increasing resistance of planktonic and biofilm cultures of Burkholderia cepacia to ciprofloxacin and ceftazidime during exponential growth. AB - The change in resistance of Burkholderia cepacia to ceftazidime and to ciprofloxacin during the exponential phase and up to the onset of stationary phase was assessed along the growth curve in batch culture. B. cepacia was grown in planktonic culture and in a biofilm on a membrane support. Resistance increased progressively during the exponential phase, being increased by ten-fold about every four generations. Bacteria grown in a biofilm were about 15 times more resistant than equivalent planktonic-grown bacteria. The growth rate was not the key factor for the development of resistance. The growth phase and the mode of growth have a fundamental impact on the susceptibility of B. cepacia towards antimicrobial agents. Bacteria growing at the same rate may differ greatly in their resistance to antimicrobial agents. PMID- 9738833 TI - Comparison of the modified Stokes' method of susceptibility testing with results obtained using MIC methods and British Society of Antimicrobial Chemotherapy breakpoints. AB - The majority of clinical microbiology laboratories in the UK use comparative disc diffusion methods based on the Stokes' method to determine antibiotic susceptibility. The technical validity of the results obtained from the modified Stokes' method of disc testing and how they relate to MIC data are not known. We studied susceptibility testing using a modified Stokes' disc diffusion method for a wide range of clinical isolates against which MICs had been determined by collaborators not involved with the disc testing evaluation. Results indicated that for 1301 organism-antibiotic combinations the number of major errors (where resistant strains were reported as sensitive) was 21/468 (4.4%) and the number of minor errors (where sensitive strains were reported as resistant) was 14/713 (1.9%) using ciprofloxacin breakpoints of 0.5 and 2 mg/L. There was good correlation between the disc susceptibility test and the MIC for 119 isolates of Enterobacteriaceae tested with the exception of Serratia spp. Excluding Serratia spp. the number of major errors for Enterobacteriaceae was 1/200 (0.5%). Data revealed 2/25 (8%) major errors for Pseudomonas aeruginosa and 1/45 (2.2%) for Acinetobacter spp. Haemophilus influenzae showed a number of unexpected categorization errors. The modified Stokes' method performed accurately for Staphylococcus aureus and coagulase-negative staphylococci when tested for susceptibility to gentamicin, erythromycin, teicoplanin and vancomycin. No major errors were reported for Streptococcus pneumoniae and beta-haemolytic streptococci. Problems occurred with the detection of antibiotic resistance in Enterococcus spp. Major errors were seen for ampicillin (2/12 strains), teicoplanin (5/6 strains) and vancomycin (5/13 strains) using a 30 microg disc but only 1/13 strains using a 5 microg disc. Overall, from our data, the modified Stokes' disc diffusion antibiotic susceptibility test showed an unacceptable number of major errors but an acceptable number of minor errors. PMID- 9738834 TI - In-vitro investigation of the antibacterial activity of agents which may be used for the oral treatment of lung infections in CF patients. AB - The effect of triple therapy with ciprofloxacin, trimethoprim and either sulphadiazine or sulphamethoxazole on the MICs and development of resistance of three strains of Pseudomonas aeruginosa, two strains of Staphylococcus aureus and one of Burkholderia cepacia was compared with that of single or dual therapy with these agents using an agar dilution method. Ciprofloxacin MICs were 0.2-0.8 mg/L for the P. aeruginosa and S. aureus strains. For trimethoprim the MIC ranges were 64-128 and 0.25-1 mg/L for P. aeruginosa and S. aureus, respectively. For the sulphonamides the ranges were 64-2500 and 20-39 mg/L for P. aeruginosa and S. aureus, respectively. All combinations of agents were effective at lower concentrations than the single agents. The combination of ciprofloxacin, sulphonamide and trimethoprim showed enhanced activity against all test organisms. The highest ciprofloxacin concentration was one-tenth of the normally attainable serum concentration of 2.5 mg/L. Thus peak plasma concentrations of > or =8 x MIC for ciprofloxacin against P. aeruginosa and S. aureus are theoretically achievable in the presence of clinically acceptable concentrations of trimethoprim and a sulphonamide, making the development of resistance less likely. The development of resistance, as shown by the proportional increase in MICs, was repressed by the triple regimen as compared with the development of resistance to agents used singly or in pairs. Killing curve determinations also demonstrated the advantage of the triple-agent therapy against all organisms tested: the combination of ciprofloxacin 0.5 mg/L, trimethoprim 1 mg/L and sulphadiazine 20 mg/L had an initial bactericidal effect against log-phase inocula of 10(6) cfu/mL of two clinical strains of P. aeruginosa and one clinical strain of S. aureus. The pseudomonas strains were reduced by 2-4 log cycles. Both recovered over 24 h but did not exceed the initial inoculum. The S. aureus was reduced to 10(2) cfu/mL in 4 h and did not recover over 24 h. A repeat dose of the triple therapy against the more resistant of the P. aeruginosa strains after 12 h also had a bactericidal effect. These data suggest the possibility of an effective exclusively oral therapy for the treatment of lung infections in cystic fibrosis patients. PMID- 9738835 TI - The effects of increasing levels of quinolone resistance on in-vitro activity of four quinolones. AB - A panel of 266 clinically isolated Gram-positive cocci and Gram-negative bacilli with varying levels of resistance to ciprofloxacin were analysed for susceptibility to Du-6859a, ciprofloxacin, ofloxacin, temafloxacin and nalidixic acid. Staphylococci were divided into ciprofloxacin-susceptible, moderately resistant and highly resistant subgroups. Du-6859a was the most potent quinolone against all taxa. As ciprofloxacin resistance increased to high levels, MICs of all quinolones increased but Du-6859a MICs increased least, and ciprofloxacin MICs increased most. Less susceptible single-step mutants were selected from 80% of 15 representative clinical isolates exposed to ciprofloxacin, 71% of isolates exposed to temafloxacin, 67% of isolates exposed to Du-6859a and 53% of isolates exposed to ofloxacin. Du-6859a inhibited more mutants (67%) at a concentration of 1 mg/L than did the other quinolones (26-43%) at their susceptible breakpoints. Du-6859a was the most rapidly bactericidal quinolone in time-kill studies with Enterococcus faecalis and Enterococcus faecium. This study indicated that Du 6859a is more potent than the comparator quinolones, is less affected by the mechanisms responsible for high-level quinolone resistance and may be less likely to select resistant mutants if it has a susceptible breakpoint of 1 mg/L. PMID- 9738836 TI - Glycopeptide tolerance in Staphylococcus aureus. AB - Treatment failures with vancomycin prompted us to investigate the phenomenon of tolerance to glycopeptides in recent clinical isolates of Staphylococcus aureus. We used both MBC/MIC determinations and time-kill measurements to study tolerance to vancomycin and teicoplanin in 35 blood or heart valve isolates of S. aureus from patients with endocarditis or bacteraemia. There was generally good agreement between vancomycin tolerance indicated by an MBC:MIC ratio of > or =32 and by < or =90% kill after 6 h incubation in the presence of 20 mg/L vancomycin. However, two isolates were tolerant according to their MBC:MIC ratios but non tolerant as judged by time-kill measurements. Seven of 15 methicillin-resistant S. aureus (MRSA) isolates but only two of 20 methicillin-susceptible ones were tolerant as judged by time-kill experiments (chi2 = 4.27 with Yates' correction, P = 0.04). Seven of the 16 isolates from patients with endocarditis were tolerant, compared with only two of the 19 isolates from patients with other conditions (chi2 = 3.43 with Yates' correction, P = 0.06). Within the endocarditis and non-endocarditis subgroups, tolerance was associated more frequently with methicillin resistance than with susceptibility, but the numbers were too small for the differences to be statistically significant. Most of the vancomycin-tolerant isolates were also tolerant to teicoplanin. We conclude that glycopeptide tolerance is a real phenomenon in S. aureus, particularly amongst MRSA isolates, and can be reliably determined by our method of time-kill analysis. Tolerance may compromise glycopeptide therapy of serious S. aureus infection and should be taken into account when deciding treatment. PMID- 9738837 TI - Activated cell-wall synthesis is associated with vancomycin resistance in methicillin-resistant Staphylococcus aureus clinical strains Mu3 and Mu50. AB - We have previously reported methicillin-resistant Staphylococcus aureus clinical strains, Mu50 and Mu3, representing two categories of vancomycin resistance: Mu50 representing vancomycin-resistant S. aureus (VRSA) with MICs > or = 8 mg/L, and Mu3 representing hetero-VRSA with MICs < or = 4 mg/L using standard MIC determination methods. The mechanisms of vancomycin resistance in these strains were investigated. These strains did not carry the enterococcal vancomycin resistance genes, vanA, vanB, or vanC1-3, as tested by PCR using specific primers. However, both strains produced three to five times the amount of penicillin-binding proteins (PBPs) 2 and 2' when compared with vancomycin susceptible S. aureus control strains with or without methicillin resistance; the amounts of PBP2 produced in Mu3 and Mu50 were comparable to those in the vancomycin-resistant S. aureus mutant strains selected in vitro. Incorporation of 14C-labelled Nacetyl-glucosamine into the cell was three to 20 times increased in Mu50 and Mu3, and release of the radioactive cell wall material was increased in Mu3 (and also in Mu50, though to a lesser extent), compared with control strains. The amounts of intracellular murein monomer precursor in these strains were three to eight times greater than those found in control strains. Transmission electron microscopy showed a doubling in the cell wall thickness in Mu50 compared with the control strains. Mu3 did not show obvious cell wall thickening. These data indicate that activated synthesis and an increased rate of cell wall turnover are common features of Mu3 and Mu50 and may be the prerequisite for the expression of vancomycin resistance in S. aureus. PMID- 9738838 TI - The effect of a component of tea (Camellia sinensis) on methicillin resistance, PBP2' synthesis, and beta-lactamase production in Staphylococcus aureus. AB - Extracts of tea (Camellia sinensis) can reverse methicillin resistance in methicillin-resistant Staphylococcus aureus (MRSA) and also, to some extent, penicillin resistance in beta-lactamase-producing S. aureus. These phenomena are explained by prevention of PBP2' synthesis and inhibition of secretion of beta lactamase, respectively. Synergy between beta-lactams and tea extracts were demonstrated by disc diffusion, chequerboard titration and growth curves. Partition chromatography of an extract of green tea on Sephadex LH-20 yielded several fractions, one of which contained a virtually pure compound that showed the above-mentioned activities, at concentrations above about 2 mg/L. The observed activities are novel and distinct from the previously reported direct antibacterial activity of tea extracts. Prevention of PBP2' synthesis offers an interesting possible new approach for the treatment of infections caused by MRSA. PMID- 9738839 TI - In-vitro susceptibility of Cryptococcus neoformans isolates to fluconazole and itraconazole. AB - The in-vitro susceptibilities of 143 isolates of Cryptococcus neoformans, collected from British patients between 1994 and 1996, to fluconazole and itraconazole were compared with those of 36 isolates collected between 1971 and 1985, 41 isolates collected between 1986 and 1989, and 43 Ugandan isolates collected in 1996. Testing was done with a broth microdilution method in YNB medium supplemented with glucose, incubation at 30 degrees C for 72 h, and an endpoint of 50% inhibition. The results showed that the MIC ranges, MIC50s and MIC90s of fluconazole and itraconazole for C. neoformans isolates from the UK have remained unchanged despite the recent widespread use of triazoles for long term maintenance of patients with AIDS-associated cryptococcal meningitis. The MIC ranges, MIC50s and MIC90s of the 43 isolates from untreated Ugandan patients with AIDS were similar to those of the British isolates. Examination of our records for 1994-96 revealed six cases in which a four-fold or greater rise in the MIC of fluconazole was associated with relapsed cryptococcal meningitis. PMID- 9738840 TI - The effect of dicloxacillin and fusidic acid on the extracellular and intracellular killing of Staphylococcus aureus. AB - The effect of dicloxacillin and fusidic acid used alone and in combination on the extracellular and intracellular killing of four isolates of Staphylococcus aureus in the presence of serum was studied. At the extracellular level, dicloxacillin (8 mg/L) had a bactericidal effect on all four isolates, whereas fusidic acid (64 mg/L) had a bacteriostatic effect on two isolates and no effect on the two other isolates. Fusidic acid significantly inhibited the extracellular bactericidal effect of dicloxacillin on two isolates. Intracellular killing was measured in human neutrophil granulocytes. Dicloxacillin (8 mg/L) significantly increased the intracellular killing of all four isolates, while fusidic acid (64 mg/L) significantly increased the intracellular killing of three isolates, but the killing was significantly lower than that of dicloxacillin. When the antibiotics were combined the intracellular killing of three of the isolates was significantly lower than that of dicloxacillin alone. The viability of the granulocytes and their ability to produce superoxide anion were not affected by the antibiotics. In conclusion, we found that the increased intracellular killing of S. aureus by dicloxacillin was inhibited by fusidic acid. PMID- 9738841 TI - Bacterial concentrations in pus and infected peritoneal fluid--implications for bactericidal activity of antibiotics. AB - Little is known about how many bacteria are present at an infectious focus at the onset of antibiotic therapy. The number of cfu was determined in pus and infected peritoneal fluids obtained from 41 patients. Pathogens were detected in 71% of specimens. There were high concentrations of bacteria in culture-positive samples, in both soft-tissue and peritoneal infections, averaging 2 x 10(8) cfu/mL. These concentrations were much higher than the standard inoculum size used in in-vitro susceptibility tests, 5 x 10(5) cfu/mL. The impact of this discrepancy on antibacterial efficacy was studied with amikacin, ciprofloxacin, imipenem and piperacillin against Escherichia coli and Staphylococcus aureus. The inhibitory and bactericidal activities of amikacin and ciprofloxacin determined with high inocula were two to four times lower than with standard inocula, whereas the activity of piperacillin was diminished at least 128-fold. Similar activity was observed with these drugs in Mueller-Hinton broth and peritoneal fluid. The bactericidal activity of imipenem was reduced in peritoneal fluid. Thus, conditions prevailing at the infection site may compromise antibiotic activity determined in vitro. PMID- 9738842 TI - Efficacy and safety of teicoplanin plus rifampicin in the treatment of bacteraemic infections caused by Staphylococcus aureus. AB - An open study was carried out on 16 patients with hospital-acquired, bacteraemic Staphylococcus aureus infections to evaluate the safety and efficacy of teicoplanin plus rifampicin. Patients received teicoplanin 400 mg bd for the first 24 h followed by 400 mg od thereafter, and rifampicin 600 mg bd. Both agents were given intravenously. Serum samples were collected to determine trough and peak antibiotic concentrations. The MIC of teicoplanin and rifampicin and the MBC of teicoplanin were determined for all S. aureus isolates. Time-kill curves were performed for the drugs individually and in combination. Clinical efficacy was assessed by the APACHE II scoring system. Bacteriological success was evaluated by elimination, persistence or recurrence of S. aureus. Safety was carefully monitored by regular biochemical and haematological testing and recording of adverse events. Fifteen patients were evaluable, of whom 13 (86.7%) were clinically cured with elimination of S. aureus. One patient died, but death was not attributed to the study drugs. Treatment failed in another patient who relapsed with a high fever. S. aureus was recovered from blood cultures from this patient, and resistance to rifampicin had developed. Time-kill curves all showed adequate killing of S. aureus at the drug concentrations measured in vivo. Neither synergy nor antagonism between teicoplanin and rifampicin was demonstrated. The combination of teicoplanin and rifampicin is an effective and well-tolerated treatment for bacteraemic S. aureus infections, but in deep-seated foci of infection resistance to rifampicin may develop. PMID- 9738843 TI - Diethylcarbamazine-related antimicrobial activity in Mycobacterium tuberculosis infected blood. AB - Previous studies have suggested that diethylcarbamazine (DEC), a drug used for filariasis control, may be useful in the treatment of mycobacterial infections. In this experiment, Mycobacterium tuberculosis was added to blood samples from two groups (healthy and diabetic) of adult non-smoking donors. Portions of each sample were tested with and without DEC at clinically achievable levels. Statistically significant DEC-related percentage decreases in BACTEC growth index counts were noted for each group (Wilcoxon one-sample signed-rank tests, alpha = 0.05, two-tailed). These results suggest that administration of DEC for filariasis control could have a positive impact on tuberculosis control. PMID- 9738844 TI - In-vitro antibiotic susceptibility and molecular analysis of anaerobic bacteria isolated in Cape Town, South Africa. AB - One hundred and twenty-three anaerobic isolates from Cape Town, South Africa, were tested in vitro against cefoxitin, chloramphenicol, metronidazole, co amoxiclav, benzylpenicillin and clindamycin. Forty-five Gram-positive organisms were tested against RP59500 (a streptogramin). Resistance in the Gram-positive isolates was as follows: of the Clostridium perfringens isolates, 4% were resistant to benzylpenicillin and 4% to clindamycin; of the Peptostreptococcus anaerobius isolates, 10% were resistant to benzylpenicillin, 10% to cefoxitin and 10% to metronidazole; of the isolates of Peptostreptococcus spp., 12% were resistant to benzylpenicillin, 6% to metronidazole, 6% to chloramphenicol and 12% to RP59500. Of the Gram-negative organisms, those in the Bacteroides fragilis group were resistant to benzylpenicillin (83%), cefoxitin (5%), clindamycin (5%) and co-amoxiclav (2%). One clindamycin-resistant B. fragilis isolate carried a plasmid homologous to the ermF erythromycin resistance determinant. PMID- 9738845 TI - Sub-MICs of sanfetrinem promote the interaction of human polymorphonuclear granulocytes with a multiply resistant strain of Klebsiella pneumoniae. AB - The effect of sanfetrinem, a member of a new class of antibiotics, upon the in vitro interaction between human polymorphonuclear granulocytes (PMNs) and a multiply resistant strain of Klebsiella pneumoniae was investigated. Sub-MICs of sanfetrinem significantly enhanced PMN phagocytosis and greatly reduced the survival of intracellular bacteria compared with antibiotic-free systems. The distinction between any effect of sanfetrinem on the klebsiellae and the phagocytes was made by exposing each of them to the drug before they were incubated together. The results indicate that sanfetrinem may have a direct positive action either on K. pneumoniae or on human granulocytes. PMID- 9738846 TI - Voriconazole against fluconazole-susceptible and resistant candida isolates: in vitro efficacy compared with that of itraconazole and ketoconazole. AB - We compared the in-vitro activity of fluconazole, itraconazole, ketoconazole and voriconazole against 67 blood isolates of Candida spp. exhibiting a wide range of fluconazole MICs (0.125 to >64 mg/L). Voriconazole was the most potent in vitro, followed by itraconazole, ketoconazole and fluconazole. Itraconazole and voriconazole had in-vitro activity against fluconazole-susceptible and -resistant candida isolates. Higher itraconazole and voriconazole MICs were observed in isolates exhibiting higher fluconazole MICs, suggesting cross-resistance. Itraconazole and voriconazole MICs of > or =16 mg/L were observed only in Candida albicans and Candida tropicalis. Candida krusei and Candida glabrata exhibited itraconazole MICs of 0.5-1 mg/L and voriconazole MICs of 0.25-0.5 mg/L. PMID- 9738847 TI - Comparison of four antibiotics in a murine model of necrotizing cutaneous infections caused by toxigenic Streptococcus pyogenes and Staphylococcus aureus. AB - The ability of azithromycin, erythromycin, clarithromycin, or cefuroxime to modify the course of group A streptococcus (GAS) or Staphylococcus aureus soft tissue infection was compared in a mouse model. In GAS-infected mice given azithromycin, fewer demonstrated dermonecrosis (P = 0.0004); the average weight gain was greater (P < 0.05) and the latency to sustained weight gain was shorter (P < 0.05) than for animals given other antibiotics. All antibiotics were effective against S. aureus infections, with no significant differences among treatments in parameters evaluated. The effectiveness of azithromycin in GAS infected mice may be related to the high and sustained tissue concentrations achieved with this antibiotic. PMID- 9738848 TI - Comparative grepafloxacin phototoxicity in mouse skin. AB - This study was performed in order to compare the phototoxic potential of grepafloxacin (a new fluoroquinolone for the treatment of respiratory tract infections) with that of a number of marketed fluoroquinolones. Groups of Balb/c mice received either control material or grepafloxacin, lomefloxacin, sparfloxacin, ofloxacin, ciprofloxacin or enoxacin at an oral dose of 200 mg/kg before being exposed to 20 J/cm2 longwave ultraviolet irradiation for 110-115 min. Lomefloxacin and sparfloxacin caused erythema and oedema which were often severe and lasted the full 7 days of the study. Enoxacin caused a long-lasting erythema, while the erythema seen following the administration of grepafloxacin, ciprofloxacin and ofloxacin was relatively mild and short-lived. The results of this study demonstrate the good safety profile of grepafloxacin in terms of phototoxicity. PMID- 9738849 TI - Current MIC breakpoints may understate the potential efficacies of carbapenems for treatment of patients with infections caused by strains of Streptococcus pneumoniae that are resistant or of intermediate susceptibility to penicillin. PMID- 9738850 TI - Study on the in-vitro activity of LY333328 against gram-positive cocci. PMID- 9738851 TI - Activities of cefepime and five other antibiotics against nosocomial PER-1-type and/or OXA-10-type beta-lactamase-producing Pseudomonas aeruginosa and Acinetobacter spp. PMID- 9738852 TI - Evaluation of the activities of two-drug combinations of rifampicin, polymyxin B and ampicillin/sulbactam against Acinetobacter baumannii. PMID- 9738853 TI - A study of the mechanisms involved in imipenem resistance in Pseudomonas aeruginosa isolates from Japan. PMID- 9738854 TI - Emergence of resistance to third-generation cephalosporins amongst Salmonella typhimurium isolates in Greece: report of the first three cases. PMID- 9738855 TI - Isolation of glycopeptide resistant Streptococcus gallolyticus strains with vanA, vanB, and both vanA and vanB genotypes from faecal samples of veal calves in The Netherlands. PMID- 9738856 TI - Activity of nisin against Streptococcus pneumoniae, in vitro, and in a mouse infection model. PMID- 9738857 TI - An isocratic high performance liquid chromatography (HPLC) assay for moxifloxacin, a new 8-methoxyquinolone. PMID- 9738858 TI - Volumetric neuroimaging in Usher syndrome: evidence of global involvement. AB - Usher syndrome is a group of genetic disorders consisting of congenital sensorineural hearing loss and retinitis pigmentosa of variable onset and severity depending on the genetic type. It was suggested that the psychosis of Usher syndrome might be secondary to a metabolic degeneration involving the brain more diffusely. There have been reports of focal and diffuse atrophic changes in the supratentorial brain as well as atrophy of some of the structures of the posterior fossa. We previously performed quantitative analysis of magnetic resonance imaging studies of 19 Usher syndrome patients (12 with type I and 7 with type II) looking at the cerebellum and various cerebellar components. We found atrophy of the cerebellum in both types and sparing of cerebellar vermis lobules I-V in type II Usher syndrome patients only. We now have studied another group of 19 patients (with some overlap in the patients studied from the previous report) with Usher syndrome (8 with type I, 11 with type II). We performed quantitative volumetric measurements of various brain structures compared to age- and sex-matched controls. We found a significant decrease in intracranial volume and in size of the brain and cerebellum with a trend toward an increase in the size of the subarachnoid spaces. These data suggest that the disease process in Usher syndrome involves the entire brain and is not limited to the posterior fossa or auditory and visual systems. PMID- 9738859 TI - Craniofacial dyssynostosis with cryptorchidism and normal stature. AB - We describe an Arab boy with craniofacial dyssynostosis. He presented with facial anomalies, mental retardation, epilepsy, hypotonia, and agenesis of the corpus callosum. This report reemphasises the previously reported traits of craniofacial dysostosis syndrome and suggests that cryptorchidism represents part of the syndrome profile and that the presence of normal stature does not preclude the diagnosis. PMID- 9738860 TI - Craniofacial dyssynostosis: a further case report. AB - I describe a boy with lambdoid craniosynostosis, severe global developmental delay, epilepsy, oculomotor dyspraxia, very thin corpus callosum, and minor anomalies. The phenotype is in keeping with a diagnosis of craniofacial dyssynostosis. This autosomal recessive condition was first described in 1976 and was originally predicted to be relatively common in those of Spanish descent. However, there have been no further reports of the condition. This case is remarkably similar to those previously described, but has the additional finding of a neuronal migration defect. PMID- 9738861 TI - Crigler-Najjar syndrome in Saudi Arabia. AB - Crigler-Najjar (CN) syndrome is a congenital familial nonhemolytic jaundice associated with high level of unconjugated bilirubin due to deficient uridine diphosphate glucuronosyltransferase (UDPG-T) activity in the liver. The aim of this report is to emphasize the need for increased awareness of this potentially fatal condition unless diagnosed early and managed appropriately. Between 1986 1994, 12 patients (8 males and 4 females) were diagnosed at our hospital with CN syndrome. Jaundice was detected in the first few days of life in all but one, in whom detection was delayed for two weeks and resulted in kernicterus. Exchange transfusions were necessary in six cases. Consanguinity was present in 11 patients, eight of whom were the offspring of first cousins. None of the patients responded to phenobarbital therapy alone, which reflects the severity of their disease. Six patients required only phototherapy while the remaining six patients required a combination of phenobarbital and phototherapy. Percutaneous liver biopsy, performed in 10 patients, showed minimal and focal cholestasis in eight, while the remaining two had a normal histological picture. Almost complete absence of the activity of UDPGT in the liver was reported in seven cases. Kernicterus developed in five cases. It is concluded that CN syndrome remains a potentially fatal condition unless diagnosed early and managed appropriately. The recent adoption of liver segment transplantation, whether orthotopic or living related, has saved affected patients the daily long hours of phototherapy. One of our patients successfully underwent living-related segmental liver transplantation. PMID- 9738862 TI - Bartsocas-Papas syndrome in an Arab family with four affected sibs: further characterization. AB - Bartsocas-Papas syndrome is a severe autosomal recessive popliteal pterygium syndrome. Other anomalies include microcephaly, facial clefts, filiform bands, ankyloblepharon, syndactyly, and other ectodermal and nonectodermal anomalies. We report on four Arab sibs with manifestations of this syndrome and some additional traits that include cutis aplasia, widely spaced nipples, low-set umbilicus, and unilateral renal hypoplasia among others. One was stillborn, and the other three children lived 10-17 months. Parents were nonconsanguineous, derived from different Bedouin tribes in Qatar and the United Arab Emirates. Similar cases from the literature are reviewed. PMID- 9738863 TI - Recurrent missense mutation in the protoporphyrinogen oxidase gene underlies variegate porphyria. AB - The porphyrias represent a heterogeneous group of disorders of porphyrin or porphyrin-precursor metabolism, resulting from the inherited or acquired dysregulation of one of the eight enzymes in the porphyrin-heme biosynthetic pathway. Variegate porphyria, one of the acute hepatic porphyrias, is characterized by a partial reduction in the activity of the penultimate enzyme in the heme biosynthetic pathway, protoporphyrinogen oxidase (PPO). Recently, VP has been linked to the PPO gene on chromosome 1q22-23, and several disease-causing mutations have been described. In this study, we identified the underlying genetic lesion in two unrelated patients with VP and investigated all available family members by polymerase chain reaction, heteroduplex analysis, automated sequencing, and restriction enzyme digestion. Mutation analyses in both families revealed a G-to-A transition in exon 6 of the PPO gene resulting in the substitution of arginine by histidine at position 168 of the protein (R168H). This arginine residue is evolutionarily conserved in human, mouse, and Bacillus subtilis, indicating the importance of this residue in PPO function. Our study establishes a recurrent missense mutation as the underlying genetic defect in two unrelated patients with VP and explains the occurrence of the phenotype in their families. PMID- 9738864 TI - Robinow syndrome, vaginal atresia, hematocolpos, and extra middle finger. AB - A 14-year-old girl with Robinow syndrome was admitted with severe abdominal pain that had recurred periodically during the last 6 months. She had been followed by us since age 2 months and she had not experienced menarche yet; hematocolpos related to vaginal atresia was diagnosed. She underwent vaginoplasty with cervical construction. Genital system abnormalities are common in Robinow syndrome, but this kind of malformation has not been reported previously. PMID- 9738865 TI - Familial complex chromosome rearrangement giving rise to balanced and unbalanced recombination products. AB - We report on a family ascertained through a 14-month-old girl with a terminal deletion of chromosome 8p23.1. Analysis of the karyotype of other relatives showed that the mother is the carrier of a balanced complex 4-break chromosome rearrangement, which she and her brother inherited from their father following recombination. This complex chromosome rearrangement (CCR) was confirmed by fluorescence in-situ hybridization (FISH) using libraries for chromosomes 1, 8, and 9, and telomeric probes for the long arm of chromosome 9. The karyotype of the maternal grandfather was 46,XY,t(1;8) (p31;q21.1),t(8;9) (p23.1;q34). The karyotype of his daughter is 46,XX,rec(8)t(1;8) (p31;q21.1)t(8;9)(p23.1;q34)pat. The karyotype of the proposita is 46,XX,rec(8)t(8;9) (p23.1;q34)mat, and that of her abnormal elder sister is 46,XX,t(1;8)(p31;q21.1)rec(8) t(8;9) (p23.1;q34)mat,der(9)t(8;9) (p23.1;q34) mat. Unbalanced segregation and/or recombination during maternal meiosis gave rise to the two abnormal sisters, one effectively with 8p trisomy and the other with monosomy for that same 8p segment. To our knowledge, this is the first case of a familial CCR giving rise to unbalanced recombination products. PMID- 9738866 TI - Centric fission of chromosome 9 in a boy with trisomy 9p. AB - A centric fission of chromosome 9 was found in a boy with trisomy 9p resulting from a de novo del (9p) and a 9p isochromosome. The patient presented with clinical findings similar to those described in previously reported cases of trisomy 9p. The cytogenetic evaluation and the molecular analysis using fluorescence in situ hybridization (FISH) with specific alphoid probe for chromosome 9 showed a karyotype of 47,XY,+del(9)(q10),+i(9p). We suggest that the mechanism leading to this situation is unusual. PMID- 9738867 TI - Sequential G-banding and fluorescent in situ hybridization on peripheral blood, bone marrow, and amniotic fluid samples. AB - We describe a method for fluorescent in situ hybridization (FISH) on previously G banded slides and mounted coverslip preparations. The technique is successful on previously G-banded slides from peripheral blood and bone marrow specimens and on amniotic fluid samples from in situ harvests. Satellite sequence probes, whole chromosome paints, and unique sequence probes have all produced reliable results. PMID- 9738868 TI - Racial and ethnic variations in the prevalence of orofacial clefts in California, 1983-1992. AB - To investigate variations in the prevalence of oral cleft anomalies according to parental race and ethnicity and maternal country of birth, the authors analyzed a cohort of 2,221,755 live births and fetal deaths delivered between 1983 and 1992 to residents of California. A total of 2,329 cleft lip with or without cleft palate (CL +/- P) cases and 1,475 cleft palate alone (CP) cases were identified by the California Birth Defects Monitoring Program, a population-based registry. Compared to Whites, the prevalence of CL +/- P was lower among African Americans (prevalence ratio (PR) = 0.56, 95% confidence interval (CI) = 0.45-0.69), higher among Native Americans (PR = 1.81, CI = 1.20-2.69), and the same among the Japanese (PR = 1.07, CI = 0.62-1.82) and Chinese (PR = 0.96, CI = 0.71-1.29). The risk of CL +/- P was slightly lower among the offspring of foreign-born Chinese women relative to U.S.-born Chinese women (PR = 0.71, CI = 0.33-1.57), and slightly higher among foreign-born Filipinos relative to their U.S.-born counterparts (PR = 1.37, CI = 0.57-3.53), although confidence intervals around these risk estimates were wide owing to sparse data. For CP, lower prevalences were observed among African Americans (PR = 0.72, CI = 0.58-0.91) and Hispanics (PR = 0.77, CI = 0.67-0.87) than among Whites. The risk of CP was higher among foreign-born Filipinos compared to U.S.-born Filipinos (PR = 1.52, CI = 0.58 4.33), although the confidence interval around this estimate included unity. These prevalence variations may reflect differences in both environmental and genetic factors affecting clefting risk. PMID- 9738869 TI - An efficient, robust, and unified method for mapping complex traits (II): multipoint linkage analysis. AB - Extending the method for two-point linkage analysis [Zhao et al., 1998: Am J Med Genet 77:366-383], this paper introduces a semiparametric method for multipoint linkage analysis, expected to gain efficiency by using multiple markers simultaneously. Overcoming the longstanding statistical and computational challenge to the parametric approaches (or lod score methods) for multipoint linkage analysis, this semiparametric approach, based on the estimating equation technique, yields statistically efficient and yet robust estimates and enjoys the computational efficiency in processing multiple markers from large pedigrees. Its computational burden increases linearly with the sizes of pedigrees and with the number of marker loci. To illustrate this semiparametric method, we apply it to marker data gathered for the Breast Cancer Consortium. The result supports the earlier finding of the positive linkage with BRCA1 and has also shown that the multipoint linkage analysis has an improved power. In addition, we have applied this method to analyze genome scanning data that have been used to localize genes responsible for type 1 diabetes. In support of the earlier findings, the genome scanning detects the linkage signals on chromosome 6 but does not support the earlier suggestions of two major genes in that genome segment. Through sensitivity analysis, it appears that the results are robust to misspecification of penetrance and allele frequency. PMID- 9738870 TI - Sacral tumors in Schinzel-Giedion syndrome. PMID- 9738871 TI - Incontinentia pigmenti versus hypomelanosis of Ito: the whys and wherefores of a confusing issue. PMID- 9738872 TI - Vitamin K deficiency embryopathy. PMID- 9738873 TI - Carrier frequency for glycogen storage disease type II in New York and estimates of affected individuals born with the disease. PMID- 9738874 TI - Mutations in genes encoding extracellular matrix proteins suppress the emb-5 gastrulation defect in Caenorhabditis elegans. AB - The second division of the gut precursor E cells is lethally accelerated during Caenorhabditis elegans gastrulation by mutations in the emb-5 gene, which encodes a presumed nuclear protein. We have isolated suppressor mutations of the temperature-sensitive allele emb-5(hc61), screened for them among dpy and other mutations routinely used as genetic markers, and identified eight emb-5 suppressor genes. Of these eight suppressor genes, at least four encode extracellular matrix proteins, i.e., three collagens and one proteoglycan. The suppression of the emb-5 gastrulation defect seemed to require the maternal expression of the suppressors. Phenotypically, the suppressors by themselves slowed down early embryonic cell divisions and corrected the abnormal cell division sequence of emb-5 mutant embryos. We propose an indirect stress-response mechanism to be the main cause of the suppression because: (1) none of these suppressors is specific, either to particular temperature-sensitive emb-5 alleles or to the emb-5 gene; (2) suppressible alleles of genes, reported here or elsewhere, are temperature sensitive or weak; (3) the suppression is not strong but marginal; (4) the suppression itself shows some degree of temperature dependency; and (5) none of the extracellular matrix proteins identified here is known to be expressed in oocytes or early embryos, despite the present observation that the suppression is maternal. PMID- 9738875 TI - Cloning and characterization of Pros45, the Drosophila SUG1 proteasome subunit homolog. AB - The proteasome plays essential roles in a variety of cellular processes, including degradation of the bulk of cellular proteins, degradation of short lived proteins such as cell cycle regulators, generation of antigenic peptides, and mediating programmed cell death. One of the best characterized subunits of the 26S proteasome is encoded by the yeast gene SUG1. We report here the cloning and characterization of the Drosophila homolog of this gene, Pros45. At the protein level, Pros45 is highly conserved with respect to its homologs in a variety of taxa: it shows 74% identity to yeast Sug1; 86% to mouse m56/mSug1/FZA B; 87% to human Trip1; and 97% to moth 18-56. Using a genomic clone as a probe for in situ hyridization to polytene chromesomes, we demonstrated that Pros45 maps to 19F, near the base of the X chromosome. Use of a pros45 cDNA clone as a probe revealed a second site of hybridization at 99CD. Pros45 mRNA is found in the unfertilized egg and in all cells of the early embryo. By the end of embryogenesis, Pros45 is expressed predominantly in the central nervous system. Targeted expression of Pros45 in a variety of different cells using the Gal4 UAS P-element system failed to generate an overt phenotype. This study provides the foundation for further examination of the role of the 26S proteasome in homeostasis and development in Drosophila. PMID- 9738876 TI - Cytokinin stimulates and abscisic acid inhibits greening of etiolated Lupinus luteus cotyledons by affecting the expression of the light-sensitive protochlorophyllide oxidoreductase. AB - Plastid biogenesis in etiolated lupine (Lupinus luteus L.) cotyledons is highly sensitive to cytokinins and abscisic acid. In the presence of the synthetic cytokinin N6-benzylaminopurine, greening and plastid biogenesis is substantially promoted as compared to untreated controls, whereas abscisic acid has an inhibitory effect. Faster greening in cytokinin-treated cotyledons is accompanied by a higher level and slower degradation of the light-sensitive protochlorophyllide-oxidoreductase (POR); while ABA has the opposite effect. The phytohormones appear to modulate POR gene expression, since the steady-state levels of POR mRNA, as well as transcripts of other nuclear genes for plastid proteins, are strongly increased by cytokinin and reduced by abscisic acid treatment. When etiolated lupine cotyledons were illuminated with far-red light prior to phytohormone application, the POR level substantially decreased; this was accompanied by the loss of the phytohormone's effect on greening. Based on these findings it is concluded that the level of POR and the integrity of the prolamellar body is crucial for cytokinin- and abscisic acid-controlled greening following transfer of etiolated lupine cotyledons into the light. PMID- 9738877 TI - Ste50p is involved in regulating filamentous growth in the yeast Saccharomyces cerevisiae and associates with Ste11p. AB - STE50 is required to sustain pheromone-induced signal transduction in S. cerevisiae. Here we report that Ste50p is involved in regulating pseudohyphal development. Both of these processes are also dependent on Ste11p. Deletion of STE50 leads to defects in filamentous growth, which can be suppressed by overproduction of Ste11p. Overexpression of STE11 also suppresses the mating defects of ste50 mutants. We have analysed the physical association between Ste50p and Ste11p in extracts of cells harvested under various conditions. A Ste11p-Ste50p complex can be isolated from extracts of cells in which the pheromone response has been activated, as well as from normally growing cells. Formation of the Ste50p-Ste11p complex does not require G(alpha), G(beta), Ste20p or Ste5p. Oligomerisation of Ste11p is shown to be independent of activation of the pheromone response pathway, and occurs in the absence of Ste50p. We conclude that Ste50p is necessary for Ste11p activity in at least two differentiation programmes: mating and filamentous growth. PMID- 9738878 TI - The P22 Erf protein and host RecA provide alternative functions for transductional segregation of plasmid-borne duplications. AB - A tandem DNA duplication carried on a ColE1-derived plasmid segregates at high frequency upon generalized transduction by phage P22 HT. Transductional segregation of the plasmid-borne duplication can be promoted either by RecA or by the Erf function of P22, indicating that transductional segregation is a consequence of the recombination events that re-circularize the plasmid in the recipient cell. RecA-mediated and Erf-mediated transduction give similar frequencies of duplication segregation and yield the same types of segregation products, indicating that two distinct recombination machineries (RecA + RecBCD and Erf + RecBCD) perform similar or identical recombination reactions on plasmid DNA substrates transduced by bacteriophage P22 HT. PMID- 9738879 TI - The sfiX, rfe and metN genes of Salmonella typhimurium and their involvement in the His(c) pleiotropic response. AB - Two loci involved in the pleiotropic response of His(c) strains of Salmonella typhimurium (sfiX and sfiY) have been characterized at the molecular level. The sfiX gene (CS 44) has been identified as a homolog of the E. coli gene sanA, located downstream of the cytidine deaminase gene (cdd). The cdd-sanA (or cdd sfiX) operon shows a highly conserved structure in E. coli and Salmonella. Like its E. coli homolog, the sfiX gene of S. typhimurium is required for vancomycin resistance at high temperature. The dual effect of sfiX mutations (induction of vancomycin sensitivity and suppression of cell division inhibition) suggests a link between SfiX function and murein synthesis. The sfiY locus (CS 85), contains two genes arranged in a single transcriptional unit. The upstream gene is a homolog of the E. coli gene rfe; mutations in this gene suppress the cell division defect of His(c) strains. The suppressor effect of rfe mutations can be reproduced by tunicamycin, suggesting that suppression of filamentation results from an increase in the intracellular concentration of UDP-N-acetyl-D glucosamine. The gene located downstream of rfe is also found in E. coli but its function is unknown. Insertions in rfe suppress the methionine requirement of His(c) strains of S. typhimurium by a polar effect on the downstream gene, tentatively designated metN. Complementation with a rfe+ clone indicates that the rfe gene is not involved in the methionine requirement of His(c) strains. Thus metN expression appears to cause methionine auxotrophy in a His(c) background. PMID- 9738881 TI - Mutations in the dim-1 gene of Neurospora crassa reduce the level of DNA methylation. AB - Mutants that show reduced DNA methylation were identified in a mutant screen based on the assumptions that (i) the nucleoside analog 5-azacytidine (5-azaC) promotes the formation of potentially lethal DNA-methyltransferase adducts; (ii) reduction in DNA methyltransferase will decrease the sensitivity of cells to 5 azaC; and (iii) this potential selective advantage will be enhanced in mutants that are deficient in the repair of 5-azaC-induced DNA damage. Of fifteen potential repair mutants screened for sensitivity to 5-azaC, five (mus-9, mus-10, mus-11, mus-18, and uvs-3) showed moderately increased sensitivity and two (mus 20, mei-3) showed highly increased sensitivity. A mus-20 mutation was used to isolate three non-complementing methylation mutants. The mutations, named dim-1 (defective in methylation), reduced female fertility, reduced methylation by 40 50%, and altered patterns of methylation. In wild-type strains hypomethylation per se fails to alter methylation specificity. We demonstrate a growth-phase dependent change in methylation patterns, detectable only in hypomethylated DNA from dim+ cultures. This may represent a growth-phase-dependent change in the relative amounts of distinct species of methyltransferase, one of which may be encoded by the dim-1 gene. PMID- 9738880 TI - Induction by alkaloids and phenobarbital of Family 4 Cytochrome P450s in Drosophila: evidence for involvement in host plant utilization. AB - In vertebrates, cytochrome P450s of the CYP2 and CYP3 families play a dominant role in drug metabolism, while in insects members of the CYP6 and CYP28 families have been implicated in metabolism of insecticides and toxic natural plant compounds. A degenerate 3' RACE strategy resulted in the identification of fifteen novel P450s from an alkaloid-resistant species of Drosophila. The strong (17.4-fold) and highly specific induction of a single gene (CYP4D10) by the toxic isoquinoline alkaloids of a commonly utilized host-plant (saguaro cactus) provides the first indication that members of the CYP4 family in insects may play an important role in the maintenance of specific insect-host plant relationships. Strong barbiturate inducibility of CYP4D10 and two other D. mettleri P450 sequences of the CYP4 family was also observed, suggesting a pattern of xenobiotic responsiveness more similar to those of several vertebrate drug metabolizing enzymes than to putative vertebrate CYP4 homologs. PMID- 9738882 TI - The rpoS gene regulates OP2, an operon for the lower pathway of xylene catabolism on the TOL plasmid, and the stress response in Pseudomonas putida mt-2. AB - Operon OP2 on the Pseudomonas putida TOL plasmid encodes enzymes for m-toluate catabolism; transcription of this operon is activated by XylS in the presence of m-toluate. Because transcriptional activation of OP2 specifically occurs in the stationary phase of growth both in P. putida and in Escherichia coli, we suspected that its transcription is dependent on RpoS (sigmaS). Therefore, we constructed a rpoS disruption strain of P. putida mt-2, and assayed OP2 expression and other phenotypes. OP2 transcription was dependent on rpoS, indicating that the stationary-phase specific activation of OP2 is due to positive regulation by RpoS in P. putida mt-2. The rpoS mutant exhibited reduced viability during the stationary phase and was sensitive to high salt concentrations and H2O2. P. putida mt-2 has two catalase isozymes, KatA and KatB. Expression of the katB gene was specific to the stationary phase and entirely dependent on the rpoS gene, while the katA gene expressed during log phase partially required rpoS. There were no significant changes in tolerance to high temperature or UV light in the rpoS mutant. No difference was observed between the E. coli rpoS mutants and their parents in their capacity to activate OP2, suggesting that the mechanism of OP2 activation in the stationary phase of growth differs between P. putida and E. coli. PMID- 9738883 TI - P element sequences can compensate for a deletion of the yellow regulatory region in Drosophila melanogaster. AB - The effects of interactions between P element and yellow regulatory sequences on the control of yellow expression were studied. The y mutations used in the analysis lack a segment of upstream sequence that extends from position -146 bp to -70 bp, relative to the transcription start site of the yellow gene. This sequence has been found to be necessary for the function of the yellow promoter. The insertion of one or two P element copies at position -69 bp compensates for the deletion in the regulatory region and restores yellow expression. After mobilization of the P element, new phenotypes were selected and molecularly characterized. Two regions in the 5' part of the P element, from 23 bp to 71 bp and from 82 bp to 108 bp, can each partially compensate for the yellow deletion. In addition, deletion derivatives of the P element were themselves able to activate yellow transcription. All such P elements retain at least 108 bp of sequence at the 5' end and 15-17 bp at the 3' end. Thus, the region of the P element from 23 bp to 108 bp contains cis-regulatory elements that can influence the transcription of neighboring genes. PMID- 9738884 TI - Genetic evidence that the two isozymes of sucrose synthase present in developing maize endosperm are critical, one for cell wall integrity and the other for starch biosynthesis. AB - In maize, two paralogous genes, Sh1 and Sus1, encode two biochemically similar isozymes of sucrose synthase, SS1 and SS2, respectively. Previous studies have attributed the mild starch deficiency of the shrunken1 (sh1) endosperm to the loss of the SS1 isozyme in the mutant. Here we describe the first mutation in the sucrose synthase1 (Sus1) gene, sus1-1, and the isolation of a double recessive genotype, sh1 sus1-1. Combined data from diverse studies, including Northern and Western analyses, RT-PCR and genomic PCR, cloning and sequencing data for the 3' region, show that the mutant sus1-1 gene has a complex pattern of expression, albeit at much reduced levels as compared to the Sus1 gene. Endosperm sucrose synthase activity in sh1 sus1-1 was barely 0.5% of the total activity in the Sh1 Sus1 genotype. Significantly, comparative analyses of Sh1 Sus1, sh1 Sus1 and sh1 sus1-1 genotypes have, for the first time, allowed us to dissect the relative contributions of each isozyme to endosperm development. Starch contents in endosperm of the three related genotypes were 100, 78 and 53%, respectively. Anatomical analyses, which confirmed the previously described early cell degeneration phenotype unique to the sh1 Sus1 endosperm, revealed no detectable difference between the two sh1 genotypes. We conclude that the SS1 isozyme plays the dominant role in providing the substrate for cellulose biosynthesis, whereas the SS2 protein is needed mainly for generating precursors for starch biosynthesis. PMID- 9738885 TI - Identification, characterization, and chromosomal organization of the ftsZ gene from Brevibacterium lactofermentum. AB - The ftsZ gene was cloned from the chromosomal DNA of Brevibacterium lactofermentum by the polymerase chain reaction (PCR) using two oligonucleotides designed from two conserved regions found in most of the previously cloned and sequenced ftsZ genes from other microorganisms. ftsZ is a single-copy gene in corynebacteria and is located downstream from ftsQ and murC, indicating linkage between genes involved in peptidoglycan synthesis (mur genes) and genes involved in cell division (fts genes). The organisation of the cluster is similar to that in Streptomyces and different from those of Escherichia coli or Bacillus subtilis because ftsA is not located upstream of ftsZ. The gene was expressed in E. coli using the T7 expression system; the calculated molecular weight of the expressed protein was 50 kDa. Expression of the B. lactofermentum ftsZ gene in E. coli inhibited cell division and led to filamentation. The ftsZ gene of this organism does not complement ftsZ mutations or deletions in E. coli, when cloned on low or high-copy-number vectors. PMID- 9738886 TI - Identification and characterization of the Staphylococcus carnosus nitrate reductase operon. AB - Physiological and genetic characterization of Staphylococcus carnosus nitrate reductase-negative mutants led to the identification of the nitrate reductase operon, narGHJI. Transcription from the nar promoter was stimulated by anaerobiosis, nitrate, and nitrite. This is in accordance with the nitrate reductase activities determined with benzyl viologen as electron donor. However, in the presence of oxygen and nitrate, high transcriptional initiation but low nitrate reductase activity was observed. Since the alphabeta complex of the nitrate reductase formed during anaerobic growth was insensitive to oxygen, other oxygen-sensitive steps (e.g., post-transcriptional mechanisms, molybdenum cofactor biosynthesis) must be involved. The nitrate-reducing system in S. carnosus displays similarities to the dissimilatory nitrate reductases of Escherichia coli. However, in the S. carnosus nar promoter, no obvious Fnr and integration host factor recognition sites are present; only one site that is related to the E. coli NarL consensus sequence was found. Studies to determine whether the E. coli proteins NarL and Fnr are functional at the S. carnosus narGHJI promoter indicated that the promoter is not functional in E. coli. PMID- 9738887 TI - Characterization of 14 different putative protein kinase cDNA clones of the C4 plant Sorghum bicolor. AB - C4 photosynthesis is functionally dependent on metabolic interactions between mesophyll- and bundle-sheath cells. Although the C4 cycle is biochemically well understood, many aspects of the regulation of enzyme activities, gene expression and cell differentiation are elusive. Protein kinases are likely involved in these regulatory processes, providing links to hormonal, metabolic and developmental signal-transduction pathways. Here we describe the cloning and characterization of 14 different putative protein kinase leaf cDNA clones from the C4 plant Sorghum bicolor. These genes belong to three different protein kinase subfamilies: ribosomal protein S6 kinases, SNF1-like protein kinases, and receptor-like protein kinases. We report the partial cDNA sequences, mesophyll/bundle-sheath steady-state mRNA ratios, mesophyll/etiolated leaf steady state mRNA ratios, and the positions of 14 protein kinase genes on the genetic map of S. bicolor. Only three of the protein kinase genes described here are expressed preferentially in mesophyll cells as compared with the bundle-sheath. PMID- 9738888 TI - Mapping of Rpb3 and Rpb5 contact sites on two large subunits, Rpb1 and Rpb2, of the RNA polymerase II from fission yeast. AB - [Rpb1 and Rpb2] Mapping of the contact sites on two large subunits of the fission yeast Schizosaccharomyces pombe RNA polymerase II with two small subunits, Rpb3 and Rpb5, was carried out using the two-hybrid screening system in the budding yeast Saccharomyces cerevisiae. Rpb5 was found to interact with any fragment of Rpb1 that contained the region H, which is conserved among the subunit 1 homologues of all RNA polymerases, including the beta' subunit of prokaryotic RNA polymerases. In agreement with the fact that Rpb5 is shared among all three forms of eukaryotic RNA polymerases, the region H of RNA polymerase I subunit 1 (Rpa190) was also found to interact with Rpb5. On the other hand, two-hybrid screening of Rpb2 fragments from RNA polymerase II indicated the presence of an Rpb3 contact site in the region H which is conserved among the subunit 2 homologues of all RNA polymerases, including the beta subunit of prokaryotic RNA polymerases. Possible functions of the regions H in the subunits 1 and 2 are discussed. PMID- 9738889 TI - The uvsC and uvsE genes of Aspergillus nidulans are not required for the mutagenic repair of UV damage. AB - The uvsC gene of Aspergillus nidulans is a homolog of the RAD51 gene of Saccharomyces cerevisiae. However, with respect to its effects on UV mutagenesis, it differs from the yeast gene, since it seems to be required for UV mutagenesis; however, this conclusion is based only on data from resting conidia. To further clarify the functional role of the uvsC gene, we tested the UV mutability of strains bearing a uvsC mutation in resting as well as in germinating conidia, by the p-fluorophenyl-alanine resistance test. We also evaluated the mutability of the uvsE mutant which belongs to the same epistatic group. Our results show that the uvsC and uvsE genes do not have a significant role in the mutagenic UV-repair pathway. PMID- 9738890 TI - Cathepsin C from Schistosoma japonicum--cDNA encoding the preproenzyme and its phylogenetic relationships. AB - A cDNA encoding preprocathepsin C was isolated from adults of the asian blood fluke Schistosoma japonicum. The deduced amino acid sequence of S. japonicum cathepsin C comprised 458 amino acid residues; 22 NH2-terminal residues corresponding to the signal peptide, 199 residues corresponding to the propeptide and 237 COOH-terminal residues corresponding to the mature enzyme region. The amino acid sequence of this preprocathepsin showed 43% and 50% identity to that of human and rat, respectively. The preproenzyme shared only 59% identity with the sequence for a cathepsin C reported from Schistosoma mansoni, differing from it in active-site residues and in its potential N-glycosylation sites. Northern blot analysis showed that S. japonicum cathepsin C was expressed in greater quantities in female than in male parasites. Phylogenetic analysis utilizing the mature enzyme sequences of S. japonicum and other cathepsin Cs demonstrated that cathepsin Cs and cathepsin Bs shared a common ancestry. The unusually long prosegment observed in cathepsin C from S. japonicum and from other species was compared to that of cathepsin Bs and cathepsin Ls. The extension contained two blocks of residues which were highly conserved among cathepsin Cs. The COOH terminus of the prosegment exhibited a composite of features present in the prosegments of cathepsin Ls and cathepsin Bs. Most significantly, given the common ancestry of cathepsin B and cathepsin C, the prosegment of cathepsin C included ERFNIN-like motifs and other residues more characteristic of non cathepsin-B-like members of the papain superfamily such as cathepsin L. PMID- 9738891 TI - Purification and characterization of an inducible metalloprotease inhibitor from the hemolymph of greater wax moth larvae, Galleria mellonella. AB - In this paper, we report the detection, purification and characterization of the first metalloprotease inhibitor (IMPI) from invertebrates. IMPI was purified from the hemolymph of last-instar larvae of Galleria mellonella by precipitation with trichloroacetic acid and heat followed by affinity chromatography on a thermolysin-Sepharose column and gel filtration or reverse-phase high-performance liquid chromatography. For the detection of inhibitor activity, a new azocoll assay was established. IMPI was only detectable in larvae that had been injected with bacterial or fungal provocators, suggesting that it is induced nonspecifically during the humoral immune response. Injection of larvae with IMPI rendered them resistant to thermolysin, in quantities that normally would be lethal for them. IMPI was shown to be specific for metalloproteases. The molecular mass of IMPI was determined by mass spectrometry to be 8360 Da. Purified IMPI was heterogeneous, owing to different degrees of glycosylation with hexose/hexosamine and deoxyhexose residues. Ten cysteine residues were found in the molecule, and these are presumed to form five disulfide bridges. The amino terminus was blocked, but a partial amino-acid sequence starting from the thermolysin cleavage site was determined; this sequence exhibited no similarity with other known proteins, suggesting that the IMPI represents a new type of protease inhibitor. PMID- 9738892 TI - Stability of neutral trehalase during heat stress in Saccharomyces cerevisiae is dependent on the activity of the catalytic subunits of cAMP-dependent protein kinase, Tpk1 and Tpk2. AB - In Saccharomyces cerevisiae cAMP-dependent protein kinase (cAPK) is involved in nutrient sensing and growth regulation via the Ras/cAMP pathway. Target enzymes, e.g. neutral trehalase, are activated or inactivated rapidly by cAPK-mediated phosphorylation. In addition, stress-induced transcription of genes of the general stress-response, e.g. HSP12, is negatively regulated via cAPK. We have investigated the effect of low cAPK activity on the stress-induced expression of neutral trehalase Nth1p. For this purpose we used mutants (tpk1tpk2TPK3, tpk1TPK2tpk3 and TPK1tpk2tpk3) with double knockouts of the three TPK genes encoding catalytic subunits of cAPK. It is shown that the tpk1tpk2TPK3 mutant, which has very low cAPK activity, exhibits a heat-stress-induced inactivation of neutral trehalase that is not observed in tpk1TPK2tpk3, TPK1tpk2tpk3 mutants and wild-type cells. However, heat stress induces an increase in NTH1 mRNA in the tpk1tpk2TPK3 mutant. Introduction of a plasmid carrying the TPK1 or TPK2 gene into tpk1tpk2TPK3 cells restores the heat-induced increase of neutral trehalase activity. In vitro and in vivo results suggest that the heat induced inactivation of neutral trehalase is due to a reversible inactivation of Nth1p. Our data indicate that a certain level of phosphorylation is essential for maintenance of neutral trehalase activity during heat shock in S. cerevisiae. Two identical putative cAPK phosphorylation sites have been found in the sequence predicted for the Nth1p. Stabilization and activation of neutral trehalase may be regulated by these sites. Furthermore, our data suggest that the heat-stress-induced transcription of the NTH1 gene is not negatively regulated by cAPK, that the TPK genes have no effect on the glucose repression of the NTH1 gene, and that non detectable neutral trehalase activity in derepressed tpk1tpk2TPK3 cells is correlated with the reduced thermotolerance observed in this strain, similar to the heat-shock-recovery defect reported for the nth1delta mutant. PMID- 9738894 TI - The binding sites for Xenopus laevis FIII/YY1 in the first exon of L1 and L14 ribosomal protein genes are dispensable for promoter expression. AB - The genes coding for the ribosomal proteins (rp genes) L14 and L1 in the toad Xenopus laevis are contacted in the first exon by the frog protein, FIII/YY1, homolog of the human zinc-finger protein YY1, acting as repressor, activator and initiator of transcription. To investigate the functional significance of FIII/YY1 in the context of the two rp genes, the L14 region at nucleotide positions -105 to +44, including all of the first exon was linked to the chloramphenicol acetyltransferase (CAT) reporter gene; constructs with wild-type and mutated sites for FIII/YY1 were injected into nuclei of stage V-VI oocytes and analyzed for CAT activity. The same procedure was followed for constructs made with L1 sequences at nucleotide positions -17 to +1567. Mutations in the sites for FIII/YY1 did not change reporter activity, nor did overexpression of FIII/YY1 in the oocytes prior to injection with L1 and L14 constructs. Since oocytes are non-dividing cells, transfections were made of Xenopus kidney cells in culture with the same constructs and the results obtained in oocytes confirmed. PMID- 9738893 TI - Evidence that Hsc70 negatively modulates the activation of the heme-regulated eIF 2alpha kinase in rabbit reticulocyte lysate. AB - The role of the heat-shock cognate protein, Hsc70, in regulating the activity of the heme-regulated eIF-2alpha kinase (HRI) in hemin-supplemented rabbit reticulocyte lysate (RRL) in response to heat and oxidative stress was examined and compared with the effect of Hsc70 on HRI activation in response to heme deficiency. Hsc70 suppressed eIF-2alpha phosphorylation and maintained the guanine nucleotide exchange activity of eIF-2B in heme-deficient RRL and in hemin supplemented RRL exposed to elevated temperatures (42 degrees C), denatured protein (reduced carboxymethylated bovine serum albumin, RCM-BSA), oxidized glutathione or Hg2+. The ability of Hsc70 to inhibit HRI activation was mediated through its ability to inhibit the hyper-autophosphorylation of transformed HRI, which causes the hyperactivation of HRI. Maintenance of protein-synthesis rate was observed to be an unreliable indicator of the ability of Hsc70 to suppress HRI activation in response to stress. While Hsc70 completely reversed protein synthesis inhibition caused by Hg2+. Hsc70 only partially reversed translational inhibition caused by oxidized glutathione (GSSG) or heat shock. The inability of Hsc70 to fully protect protein synthesis from inhibition induced by heat shock or GSSG was due to inability of Hsc70 to protect eIF-4 E from heat-induced dephosphorylation, and its inability to protect translational elongation from GSSG-induced inhibition, respectively. Activation of HRI in heat-shocked hemin supplemented lysate correlated with a marked decrease in the pool of Hsc70 that was available to bind RCM-BSA and the loss of the interaction of Hsc70 with HRI. These observations indicate that heat shock induced the accumulation of a sufficient quantity of Hsc70 binding substrates (e.g., denatured protein) to sequester Hsc70 and inhibit the ability of Hsc70 to suppress HRI activation. Our results indicate that Hsc70 not only negatively modulates the activation of HRI in heme-deficienct RRL, but also in hemin-supplemented RRL in response to heat and oxidative stress. PMID- 9738895 TI - ERp28, a human endoplasmic-reticulum-lumenal protein, is a member of the protein disulfide isomerase family but lacks a CXXC thioredoxin-box motif. AB - We report on the isolation, sequence and a putative role of a human endoplasmic reticulum-lumenal protein, ERp28. The protein has the C-terminal retention signal KEEL and localizes to the endoplasmic reticulum (ER) as seen by subcellular fractionation and immunofluorescence studies. The protein has significant sequence similarity to members of the protein disulfide isomerase (PDI) family, although it lacks the thioredoxin box (CGHC) motif. We propose, on the basis of sequence analysis, a model of the domain structure of PDI, representing a significant extension of previously proposed models. Our results are in partial agreement with recently published NMR data [Kemmink, J., Darby, J., Dijkstra, K., Nilges, M. & Creighton, T. E. (1997) Curr. Biol. 7, 239-245] and indicate that PDI contains, in addition to the two thioredoxin folds described in previous models, two thioredoxin folds within the domains previously defined as b and b'. The thioredoxin domain of ERp28 shares a higher degree of similarity with the corresponding active and inactive domains of PDI than with other members of the PDI family, indicating that ERp28 developed from an ancient form of PDI or a PDI precursor. In contrast to Ig-heavy-chain-binding protein, human ERp28 is not induced by metabolic stress (tunicamycin). In in vitro experiments, ERp28 and calnexin precipitate with overexpressed, wild-type hepatitis B small surface antigen and with a mutated ER-retained form. This indicates that ERp28, as calnexin, may be involved in the processing of secretory proteins within the ER. PMID- 9738896 TI - Analysis of protein self-association at constant concentration by fluorescence energy transfer. AB - Fluorescence-resonance-energy transfer from subunits labelled with a fluorescence donor group to subunits labelled with a fluorescence acceptor group can be used for quantitative analysis of protein self-association. The present approach evaluates fluorescence measurements on mixtures of equimolar solutions of donor labelled and acceptor-labelled protein composed by systematic variation of the volume ratio. Its attractive feature is that it allows the determination of equilibrium constants at fixed total concentration. Problems encountered by most other methods, which require the equilibria to be followed to high dilution, are avoided. Conditions to be fulfilled are that a reactive site is available on the protein for specific introduction of the labels and that labelling neither affects the conformation nor interferes with the intermolecular interactions. It is desirable that the Forster distance of the donor/acceptor pair complies with its separation. While dimerisation constants can be determined exclusively by fluorescence measurements, the analysis of more complex cases of self-association depends on additional independent information. This communication reports on an application of the approach to the association/dissociation equilibrium between insulin monomers and dimers. Labelling of insulin at the epsilon-amino group of LysB29 does not disturb the conformation nor does it affect dimerisation. 2 Aminobenzoyl and 3-nitrotyrosyl residues served as the donor/acceptor pairs. Because they are less bulky than most other fluorescence labels and are of balanced polarity they do not alter the chemical nature of the protein. Their Forster distance of 29 A matches their 32-A separation in the insulin dimer. Energy transfer was measured as a function of the molar fractions of donor insulin and acceptor-insulin at constant total concentration. Evaluation of this dependence resulted in a dimerisation constant, K12, of 0.72x10(5) M(-1). Its agreement with values obtained with other methods demonstrates that the present approach is a reliable alternative. PMID- 9738897 TI - Structural analysis of ASCUT-1, a protein component of the cuticle of the parasitic nematode Ascaris lumbricoides. AB - CUT-1 from the intestinal parasitic nematode Ascaris lumbricoides is a protein component of the insoluble residue of the cuticle, cuticlin. It contains the CUT 1-like domain which is shared by members of a novel family of components of extracellular matrices. The structure and the thermal stability of recombinant CUT-1 from A. lumbricoides (ASCUT-1) were investigated by Fourier-transform infrared (FT-IR) and CD spectroscopy. The data revealed that the secondary structure of the protein at 20 degrees C, both as insoluble inclusion bodies or in soluble form, contains about 50% beta structure, 14% alpha-helix and 25% turns. A tendency of A. lumbricoides CUT-1 to form aggregates was documented by FT-IR spectroscopy which showed also that the addition of SDS disrupts these interactions. Near-ultraviolet CD spectra confirmed these data and suggested that phenylalanine residues are probably involved in intermolecular hydrophobic interactions responsible for the tendency of the protein to aggregate. Near ultraviolet spectra showed also that part of the cysteine residues forms disulphide bridges responsible for the tertiary architecture of the protein. Finally, FT-IR and CD data revealed that ASCUT-1 is very stable at high temperatures. This stability and the tendency of ASCUT-1 to form aggregates suggest that these properties may be important for a protein which is a component of a particularly resistant extracellular matrix such as the nematode cuticle. PMID- 9738898 TI - Probing of the neuropeptide Y-Y1-receptors interaction with anti-receptor antibodies. AB - The Y1 receptor, which belongs to the family of rhodopsin-like GTP-binding protein-coupled, seven-transmembrane helix-spanning receptors, binds the 36-mer neuromodulator neuropeptide Y (NPY) with nanomolar affinity. Synthetic fragments of the N-terminus, extracellular loops and C-terminus of the Y1 receptor were used to generate 18 anti-receptor antibodies; ten of them recognize the receptor expressed on intact cells as well as on membranes that have been prepared (with the exception of one antibody raised against the intracellular C-terminus) as investigated by ELISA. SDS/PAGE of solubilized membranes, subsequent Western blotting and staining with the antibodies revealed two proteins of 73 kDa and 51 kDa for both, the rat and the human receptor. Competition with neuropeptide Y showed that the binding of seven antibodies is strongly inhibited in the presence of the native ligand. Using photoactivatible analogues, it could be demonstrated that the competition efficiency strongly depends on the position of the crosslinker within the ligand. Based on these studies, a model for the ligand receptor interaction is suggested. These antibodies represent novel tools for the structural characterization of the Y1 receptor and its interaction with NPY and antagonists as well as for localization studies. PMID- 9738899 TI - Myxococcus xanthus spore coat protein S, a stress-induced member of the betagamma crystallin superfamily, gains stability from binding of calcium ions. AB - Protein S, a calcium-binding spore coat protein from the soil bacterium Myxococcus xanthus, belongs to a group of structurally related proteins, the betagamma-crystallin superfamily. Common features of this protein family are the Greek-key structural motif or crystallin fold, and the fact that all members are extremely stable long term. To investigate the correlation between the stability and Greek-key topology, protein S was cloned, expressed in Escherichia coli and purified to homogeneity. Ca2+ binding influences the native tertiary structure of protein S, whereas the secondary structure remains unaffected as shown by spectroscopic methods. Ca2+ ions enhance the conformational stability of protein S significantly. The midpoints of urea and guanidinium chloride-induced transitions show a difference of 1.4 M and 0.5 M denaturant, respectively, in the absence and in the presence of calcium. An equilibrium intermediate indicating independent domain folding can be detected at pH 2. In addition, thermal denaturation shows a clear deviation from the two-state model of folding, again with a strong stabilisation by Ca2+ ions. Temperature and denaturant-induced equilibrium transitions are fully reversible. Our data implicate a different strategy for achieving the high stability required for the biological function compared with the structurally related lens crystallins. PMID- 9738900 TI - Interaction of vanadate oligomers and permolybdate with the 90-kDa heat-shock protein, Hsp90. AB - The 90-kDa heat-shock protein (Hsp90) is a molecular chaperone that aids the folding of nuclear hormone receptors and protein kinases. Hsp90 x protein complexes can be stabilized by molybdate and by other transition metal oxyanions such as vanadate. Our earlier findings [Csermely, P., Kajtar, J., Hollosi, M., Jalsovszky, G., Holly, S., Kahn, C. R., Gergely, P. Jr, Soti, C., Mihaly, K. & Somogyi, J. (1993) J. Biol. Chem. 268, 1901-1907] showed that vanadate and molybdate can induce a large conformational change of Hsp90. Here we provide direct evidence for the binding of vanadate and molybdate to Hsp90 by demonstrating that surface-plasmon-resonance measurements indicate binding of various vanadate oligomers to Hsp90. 51V-NMR measurements show an extensive interaction of decavanadate with the chaperone, and permolybdate treatment of Hsp90 induces a marked mobility shift of the protein and its tryptic fragments. Our results indicate the flexibility of molybdate/vanadate-binding sites of Hsp90, which are able to accommodate various species of these transition metal anions. PMID- 9738901 TI - Cyclohexa-1,5-diene-1-carbonyl-CoA hydratase [corrected], an enzyme involved in anaerobic metabolism of benzoyl-CoA in the denitrifying bacterium Thauera aromatica. AB - Many aromatic compounds can be metabolized by bacteria under anoxic conditions via benzoyl-CoA as the common intermediate. The central pathway of benzoyl-CoA metabolism is initiated by an ATP-driven reduction of the aromatic ring producing cyclohexa-1,5-diene-1-carbonyl-CoA. The 1,5-dienoyl-CoA intermediate is thought to be transformed to 6-hydroxycyclohex-1-ene-1-carbonyl-CoA by a specific dienoyl CoA hydratase catalyzing the formal addition of water to one of the double bonds. This dienoyl-CoA hydratase was detected in the denitrifying bacterium Thauera aromatica after anaerobic growth with benzoate. Substrate and product were confirmed and a convenient spectrophotometric assay was developed. The equilibrium concentrations of substrate and product were almost equal. Enzyme activity was induced after anoxic growth with benzoate, in contrast to acetate. The enzyme of 28 kDa was purified from T. aromatica and was found to be highly specific for the cyclic 1,5-dienoyl-CoA. A second 29-kDa enoyl-CoA hydratase acted on crotonyl-CoA; this highly active enoyl-CoA hydratase also acted slowly on cyclohex-1-ene-1-carbonyl-CoA. The regulation of expression of dienoyl-CoA hydratase activity, the kinetic constants, the substrate specificity, and the specific activity of the enzyme in cell extract provide evidence that dienoyl-CoA hydratase is the second enzyme of the central benzoyl-CoA pathway of anaerobic aromatic metabolism in T. aromatica. Extracts of Rhodopseudomonas palustris contained high activity of cyclohex-1-ene-1-carbonyl-CoA hydratase, but no 1,5 dienoyl-CoA hydratase activity. It appears that a variant of the benzoyl-CoA pathway is operating in R. palustris in which hydration of the 1,5-dienoyl-CoA does not take place. Rather, cyclohex-1-ene-1-carbonyl-CoA is hydrated to 2 hydroxycyclohexane-1-carbonyl-CoA [corrected]. PMID- 9738902 TI - Purification and characterization of recombinant Thermotoga maritima dihydrofolate reductase. AB - We have overexpressed the gene for dihydrofolate reductase (DHFR) from Thermotoga maritima in Escherichia coli and characterized the biochemical properties of the recombinant protein. This enzyme is involved in the de novo synthesis of deoxythymidine 5'-phosphate and is critical for cell growth. High levels of T. maritima DHFR in the new expression system conferred resistance to high levels of DHFR inhibitors which inhibit the growth of non-recombinant cells. The enzyme was purified to homogeneity in the following two steps: heat treatment followed by affinity chromatography or cation-exchange chromatography. Most of the biochemical properties of T. maritima DHFR resemble those of other bacterial or eukaryotic DHFRs, however, some are unique to T. maritima DHFR. The pH optima for activity, Km for substrates, and polypeptide chain length of T. maritima DHFR are similar to those of other DHFRs. In addition, the secondary structure of T. maritima DHFR, as measured by circular dichroism, is similar to that of other DHFRs. Interestingly, T. maritima DHFR exhibits some characteristics of eukaryotic DHFRs, such as a basic pI, an excess of positively charged residues in the polypeptide chain and activation of the enzyme by inorganic salts and urea. Unlike most other DHFRs which are monomeric or part of a bifunctional DHFR thymidylate synthase (TS) enzyme, T. maritima DHFR seems to generally form a dimer in solution and is also much more thermostable than other DHFRs. It may be that dimer formation is a key factor in determining the stability of T. maritima DHFR. PMID- 9738903 TI - The third serine proteinase with chymotrypsin specificity isolated from Atlantic cod (Gadus morhua) is a type-II elastase. AB - Preparations of chymotrypsin from Atlantic cod are heterogeneous and previously gave rise to two active peaks when subjected to pH-gradient chromatography. Extension of the pH-gradient resolved a third protein peak with benzoyltyrosine ethylester hydrolytic activity. The first two peaks have been characterized as chymotrypsin variants and designated A and B, whereas the identity of the third peak was not clear. Analysis of this protein by Edman sequencing and mass spectrometry has now confirmed a high degree of identity with the predicted protein sequence from a recently described cDNA clone. That sequence was named elastase B by sequence comparison. As the present elastase deviates in 16 positions from that of elastase B, we have named it elastase C. The elastase C was active in hydrolysing typical substrates used by chymotrypsin, namely benzoyl L-tyrosine ethylester and succinyl-Ala-Ala-Pro-Phe-p-nitroanilide, but inactive against the typical elastase substrates succinyl-Ala-Ala-Ala-p-nitroanilide and orcein-elastin. Comparison of the kinetic properties of the cod elastase C with bovine chymotrypsin and cod chymotrypsin variants A and B, using succinyl-Ala-Ala Pro-Phe-p-nitroanilide, showed a lower catalytic efficiency of elastase C. The effects of several inhibitors on cod elastase C were identical to effects on chymotrypsins variants A and B, but dissimilar when compared with porcine pancreatic elastase. On the basis of the specificity and amino acid sequence, we conclude that the enzyme under study is most correctly classified as a type-II elastase. PMID- 9738904 TI - Purification and characterization of the respiratory arsenate reductase of Chrysiogenes arsenatis. AB - Chrysiogenes arsenatis is the only bacterium known that respires anaerobically using arsenate as the terminal electron acceptor and the respiratory substrate acetate as the electron donor. During growth, the arsenate is reduced to arsenite; the reduction is catalyzed by an arsenate reductase. This study describes the purification and characterization of a respiratory arsenate reductase (Arr). The enzyme consists of two subunits with molecular masses of 87 kDa (ArrA) and 29 kDa (ArrB), and is a heterodimer alpha1beta1 with a native molecular mass of 123 kDa. The arsenate reductase contains molybdenum, iron, acid labile sulfur and zinc as cofactor constituents. The Km of the enzyme for arsenate is 0.3 mM and the Vmax is 7013 micromol arsenate reduced x min(-1) x mg protein(-1). Nitrate, sulfate, selenate and fumarate cannot serve as alternative electron acceptors for the arsenate reductase. Synthesis of the protein is regulated, as arsenate must be present during growth for the enzyme to be fully induced. The N-terminus of ArrA is similar to a number of procaryotic molybdenum containing polypeptides (e.g. the formate dehydrogenases H and N of Escherichia coli). The N-terminus of ArrB is similar to iron-sulfur proteins. The respiratory arsenate reductase of C. arsenatis is different from the non-respiratory arsenate reductases of E. coli and Staphylococcus aureus. PMID- 9738905 TI - The N-terminus of A1-type myosin essential light chains binds actin and modulates myosin motor function. AB - There are two isoforms (A1 and A2) of the myosin essential light chain (ELC) and consequently two isoenzymes of myosin subfragment 1 (S1), S1(A1) and S1(A2). The two isoenzymes differ in their kinetic properties with S1(A1) having a lower apparent Km for actin and a slower turnover of MgATP (k(cat)) than S1(A2). The two forms of the ELC differ only at their N-termini where A1 has an additional 40 odd amino acids that are not present in A2. The human atrial ELC (an A1-type ELC) was overexpressed in Escherichia coli and purified by ammonium sulphate fractionation and ion-exchange chromatography. The recombinant ELC had actin activated MgATPase kinetics similar to those for rabbit skeletal S1(A1) under the same conditions. Deletion of the first 45 amino acid residues resulted in an ELC similar to the rabbit skeletal A2 isoform and, when hybridised into S1, in S1(A2) like kinetic properties. Results obtained with an ELC mutant that lacks the first 11 residues were intermediate between these two extremes but tending towards the S1(A2)-like phenotype. The wild-type ELC (both hybridised into S1 or free in solution) could be cross-linked to F-actin, whereas the deletion mutant lacking the first 45 amino acids could not. The deletion mutant lacking the first 11 amino acids cross-linked only poorly under the same conditions, consistent with the MgATPase data. We therefore conclude that these N-terminal eleven amino acids predominantly encode an actin-binding site which modulates the kinetics of the myosin motor. Furthermore, while free A1-type ELC cross-linked to both polymeric F-actin and the monomeric G-actin:DNase-I complex, the same ELC in S1(A1) could only cross-link to F-actin. This suggests that the light chain binds to a different actin monomer than the heavy chain. PMID- 9738906 TI - Functional groups of sialic acids involved in binding to siglecs (sialoadhesins) deduced from interactions with synthetic analogues. AB - The siglecs, formerly called sialoadhesins, are a family of I-type lectins binding to sialic acids on the cell surface. Five members of this family have been identified: sialoadhesin, myelin-associated glycoprotein (MAG), Schwann cell myelin protein (SMP), CD22 and CD33. We have investigated the relevance of substituents at position C-9 and in the N-acetyl group of N-acetylneuraminic acid, using a series of synthetic sialic-acid analogues either on resialylated human erythrocytes or as free alpha-glycosides in hapten inhibition. All five siglecs require the hydroxy group at C-9 for binding, suggesting hydrogen bonding of this substituent with the binding site. Remarkable differences were found among the proteins in their specificity for modifications of the N-acetyl group. Whereas sialoadhesin, MAG and SMP do not tolerate a hydroxy group as in N glycolylneuraminic acid, they bind to halogenated acetyl residues. In the case of MAG, N-fluoroacetylneuraminic acid is bound about 17-fold better than N acetylneuraminic acid. In contrast, human and murine CD22 both show good affinity for N-glycolylneuraminic acid, but only human CD22 bound the halogenated compounds. In conclusion, our data indicate that interactions of the hydroxy group at position 9 and the N-acyl substituent contribute significantly to the binding strength. PMID- 9738907 TI - Structural elucidation of the lipopolysaccharide core regions of the wild-type strain PAO1 and O-chain-deficient mutant strains AK1401 and AK1012 from Pseudomonas aeruginosa serotype O5. AB - Lipopolysaccharide (LPS) of the Pseudomonas aeruginosa serotype O5 wild-type strain PAO1 and derived rough-type mutant strains AK1401 and AK1012 was isolated by a modified phenol/chloroform/petroleum-ether extraction method. Deoxycholate/PAGE of the LPS from the rough mutant AK1401 indicated two bands near the dye front with mobilities similar to those of the parent strain, indicating that both LPS contain a complete core and a species comprising a core and one repeating unit. Composition analysis of the LPS from strains PAO1 and AK1401 indicated that the complete core oligosaccharide was composed of D-glucose (four units), L-rhamnose (one unit), 2-amino-2-deoxy-D-galactose (one unit), L glycero-D-manno-heptose (Hep; two units), 3-deoxy-D-manno-octulosonic acid (Kdo; two units), L-alanine (one unit) and phosphate (three units). The glycan structure of the LPS was determined by one-dimensional and two-dimensional (2D) NMR techniques in combination with MS-based methods on oligosaccharide samples obtained from the LPS by delipidation procedures. The locations of three phosphomonoester groups on the first heptose residue were established by a two dimensional 31P (omega1)-half-filtered COSY experiment on the reduced core oligosaccharide sample of the LPS from the wild-type strain. The presence of a 7 O-carbamoyl substituent was observed on the second heptose. The structure of the core region of the O-chain-deficient LPS from P. aeruginosa serotype 05 is as follows: [structure: see text] where R1 is beta-D-Glcp-(1-->2)-alpha-L-Rhap-(1- >6)-alpha-D-Glcp-(1--> and R2 is alpha-D-Glcp-(1-->6)-beta-D-Glcp-(1->. A structural model is presented that is also representative of that for P. aeruginosa serotype O6 LPS. A revised structure for the serotype O6 mutant strain A28 is presented. PMID- 9738909 TI - Translocation of the precursor of alpha-amylase into Bacillus subtilis membrane vesicles. AB - Bacilli vigorously secrete proteins into the extracellular environment, and are therefore used in industry for the bulk production of enzymes such as proteinases and amylases. Studies on the mechanism of protein translocation in these Gram positive bacteria have been hampered by the lack of an in vitro system. To establish such a system for Bacillus subtilis, everted membranes were isolated from a strain deficient in the alkaline and neutral protease. Translocation competent membrane vesicles were obtained only when a broad range proteinase inhibitor cocktail was used during membrane isolation. This method efficiently prevented proteolysis of SecY, one of the core integral membrane components of the preprotein translocase. Translocation of the urea-denatured precursor of the Bacillus licheniformis alpha-amylase, preAmyL, and B. subtilis alkaline phosphatase, prePhoB, into the B. subtilis membrane vesicles require the B. subtilis SecA protein and are driven by ATP hydrolysis and the proton-motive force. These studies establish an authentic in vitro translocation system for protein secretion in B. subtilis. PMID- 9738908 TI - Carboxymethylated phosphatidylethanolamine in mitochondrial membranes of mammals- evidence for intracellular lipid glycoxidation. AB - The non-enzymatic modification of aminophospholipids with lipoperoxidation derived aldehydes and glycoxidation-derived products have been reported previously. However, it remains unknown whether intracellular membranes are damaged by these glycoxidation-derived products. To investigate this issue, we tested whether aminophospholipids from mitochondrial membranes are damaged by glycoxidative stress the mitochondrion being identified as the major site of reactive-species production in the cell. We have used a selected-ion monitoring/gas-chromatography/mass-spectrometry assay for carboxymethylethanolamine (CM-Etn) detection, and provide evidence for the presence of CM-Etn in mitochondrial phospholipids. Further, as a physiological approach to evaluate the influence of mitochondrial oxidative stress in CM-Etn formation, we also present the comparative levels of CM-Etn in mitochondrial membranes of ten mammalian species ranging in maximum life-span from 3.5 years to 100, since the rate of mitochondrial reactive-oxygen-species production is inversely correlated to the maximum life-span. Our results show that CM-Etn levels correlate in a logarithmic fashion with the maximum-life-span [[CM-Etn] = 0.51 + 0.50 x', where x' = log(maximum-life-span); r = 0.81, P < 0.004]. The data demonstrate the intracellular occurrence of glycoxidative processes affecting membrane lipids. Moreover, these data show that longer-lived mammals contain higher levels of CM-Etn in mitochondrial membrane aminophospholipids. This trend could result from differences in rates of CM-Etn accumulation and/or phospholipid turnover. PMID- 9738910 TI - Transport of amino acid aryl amides by the intestinal H+/peptide cotransport system, PEPT1. AB - Transport of amino acid aryl amides by the intestinal H+/peptide symporter (PEPT1) was studied in Caco-2 cells and in Xenopus laevis oocytes expressing human PEPT1. Several amino acid amides were able to inhibit the uptake of [14C]glycylsarcosine in Caco-2 cells. Ala-4-nitroanilide (Ki = 0.08 mM), Phe-4 nitroanilide (Ki = 0.09 mM) and Ala-4-phenylanilide (Ki = 0.03 mM) were accepted as substrates with equal or higher affinity than natural Ala-Xaa dipeptides. Ala anilide (Ki = 2.9 mM), Ala-7-amido-4-methylcoumarin (Ki = 0.2 mM), Ala-4 chloroanilide (Ki = 0.3 mM) and Ala-4-methylanilide (Ki = 0.3 mM) were also recognized by PEPT1 as substrates. In contrast, alanine, Ala-amide, Phe-amide, Ala-methyl ester, Ala-4-nitrobenzyl ester and Ala-methylamide were not recognized (Ki > 20 mM). In X. laevis oocytes, transport of Ala-4-nitroanilide, Ala-7-amido 4-methylcoumarin, Ala-4-methylanilide and Ala-anilide was associated with transfer of positive charge and the currents were saturable with respect to substrate concentration (K0.5 values: 0.1, 0.2, 0.8 and 3.1 mM, respectively). The currents induced by Ala-4-methylanilide were saturable with respect to the substrate concentration and influenced by the membrane potential. The affinity of the transporter for Ala-4-methylanilide was also found to be influenced by the membrane potential. We conclude that the intestinal H+/peptide cotransport system PEPT1 accepts amino acid aryl amides as substrates. PMID- 9738911 TI - The lack of a stress response in Hydra oligactis is due to reduced hsp70 mRNA stability. AB - Synthesis and degradation of hsp70 mRNA was examined and compared in Hydra species living in different habitats and showing different heat-shock response. Hydra oligactis is restricted to habitats of low temperature and relatively stable pH. We have shown previously that this species is unable to acquire thermotolerance [Bosch, T., Krylow, S., Bode, H. & Steele, R. (1988) Proc. Natl. Acad. Sci. USA 85, 7927-7931] and synthesizes significantly less heat-shock protein and hsp70 mRNA [Gellner, K., Praetzel, G. & Bosch, T. C. G. (1992) Eur J. Biochem. 210, 683-691] in response to stress than related species, such as Hydra bulgaris or Hydra magnipapillata, which are adapted to habitats of wide temperature range and variable water quality. To examine the mechanisms responsible for the differential heat-shock responses in these species, a construct containing H. magnipapillata hsp70 regulatory sequences fused to firefly luciferase was introduced into H. oligactis and H. magnipapillata polyps, and expression of luciferase examined. The results showed that luciferase can be expressed equally well in a heat-inducible manner in both species, suggesting that H. oligactis heat-shock factor can interact with H. magnipapillata heat shock elements. Northern blots of alpha-amanitin-treated polyps demonstrated that the half-life of hsp70 mRNA in heat-shocked H. oligactis is drastically shorter than in H. magnipapillata. Thus, differences in hsp70 mRNA stability appear to be responsible for the habitat-correlated differences in the stress response in Hydra species. PMID- 9738912 TI - Substrate binding to a cyclodextrin glycosyltransferase and mutations increasing the gamma-cyclodextrin production. AB - Bacterial cyclodextrin glycosyltransferases use starch to produce cyclic maltooligosaccharides (cyclodextrins) which are of interest in various applications. The cyclization reaction gives rise to a spectrum of ring sizes consisting of predominantly six to eight glucosyl units. Using the enzyme from Bacillus circulans strain no. 8, binding studies have been performed with several substrates and analogues. The observed binding modes differ in detail, but agree in general with data on homologous enzymes. Based on these binding studies, two mutations were designed that changed the production spectrum from the predominant product beta-cyclodextrin of the wild-type enzyme towards gamma-cyclodextrin, which is of practical interest because it is rare and can encapsulate larger nonpolar compounds. PMID- 9738913 TI - Biochemical evidence for a calmodulin-stimulated calcium-dependent protein kinase in maize. AB - We provide biochemical evidence for the presence of a Ca2+-dependent calmodulin (CaM)-stimulated protein kinase (CCaMK) from etiolated maize coleoptiles. The kinase, with a molecular mass of 72.3 kDa, was purified to homogeneity by means of ammonium sulphate precipitation, DEAE-Sephacel chromatography. CaM-Sepharose chromatography and gel purification. The purified kinase required 5 mM Mg2+ for activity and had an optimum pH of 7.5. The kinase is a Ca2+-binding protein, as was evident by 45Ca2+-binding and Ca2+ mobility-gel-shift assays. 1 microM Ca2+ stimulated the kinase activity about 12-fold and was further stimulated by the addition of exogenous CaM (approximately 100 nM). Addition of Ca2+ and CaM antagonists decreased the kinase activity. Under in vitro assay conditions the kinase phosphorylated preferentially syntide-2, histone IIIS and casein. Syntide 2 and histone IIIS were phosphorylated at serine residues, showing that the kinase belongs to the serine/threonine family of protein kinases. Autophosphorylation of CCaMK occurred on threonine residue(s) and was Ca2+ dependent. Addition of exogenous CaM had no effect on autophosphorylation. The properties of the maize kinase suggests that it is a CCaMK that shows dual stimulation with Ca2+ and CaM for substrate phosphorylation and only Ca2+ requirement for autophosphorylation. Antibodies raised against the kinase cross reacted with maize total proteins to give a single band of 72 kDa and precipitated substrate (syntide-2 and histone IIIS)-phosphorylation and autophosphorylation activities in a specific manner. Localisation studies with antibodies showed that the kinase is ubiquitous. PMID- 9738914 TI - Two metal-binding sites in a lead ribozyme bound to competitively by Pb2+ and Mg2+--induced structural changes as revealed by NMR. AB - We reported recently that a lead ribozyme with modified bases cleaved at an additional site at high Pb2+ concentrations (>0.1 mM), and that the cleavage at a canonical site was enhanced nearly fourfold at the optimum combination of Pb2+ and Mg2+ concentrations [Kim, M. H., Katahira, M., Sugiyama, T. & Uesugi, S. (1997) J. Biochem. (Tokyo) 122, 1062-1067]. Here we have identified two metal binding sites (sites 1 and 2) of the lead ribozyme at the residue level by NMR. Both sites are located in an asymmetric internal loop of the lead ribozyme. Site 1 is composed of residues of an enzyme strand and site 2 of residues of a substrate strand. The two sites are bound to competitively by Pb2+ and Mg2+. It was revealed that at certain Pb2+ and Mg2+ concentrations, site 1 is occupied by Pb2+ and site 2 is occupied by Mg2+. The dependency of the cleavage at the canonical and other sites on the Pb2+ and Mg2+ concentrations is interpreted by considering the species of metal ions bound to the two sites. It is suggested that the addition of the two metal ions produces similar and different effects on the structure of the lead ribozyme, and the two metal ions have a synergistic effect on the structure. PMID- 9738915 TI - Isolation, cDNA cloning and gene expression of an antibacterial protein from larvae of the coconut rhinoceros beetle, Oryctes rhinoceros. AB - An antibacterial protein, designated rhinocerosin, was purified to homogeneity from larvae of the coconut rhinoceros beetle, Oryctes rhinoceros immunized with Escherichia coli. Based on the amino acid sequence of the N-terminal region, a degenerate primer was synthesized and reverse-transcriptase PCR was performed to clone rhinocerosin cDNA. As a result, a 279-bp fragment was obtained. The complete nucleotide sequence was determined by sequencing the extended rhinocerosin cDNA clone by 5' rapid amplification of cDNA ends. The deduced amino acid sequence of the mature portion of rhinocerosin was composed of 72 amino acids without cystein residues and was shown to be rich in glycine (11.1%) and proline (11.1%) residues. Comparison of the deduced amino acid sequence of rhinocerosin with those of other antibacterial proteins indicated that it has 77.8% and 44.6% identity with holotricin 2 and coleoptrecin, respectively. Rhinocerosin had strong antibacterial activity against E. coli, Streptococcus pyogenes, Staphylococcus aureus but not against Pseudomonas aeruginosa. Results of reverse-transcriptase PCR analysis of gene expression in different tissues indicated that the rhinocerosin gene is strongly expressed in the fat body and the Malpighian tubule, and weakly expressed in hemocytes and midgut. In addition, gene expression was inducible by bacteria in the fat body, the Malpighian tubule and hemocyte but constitutive expression was observed in the midgut. PMID- 9738916 TI - Identification of major wheat allergens by means of the Escherichia coli expression system. AB - Wheat proteins were fractionated into salt-soluble, glutenin-rich, and gliadin rich fractions. Reactivities of these protein fractions with sera of patients with wheat-associated allergies were examined under various conditions. The relative reactivity of the fractions was generally in the order glutenin-rich > gliadin-rich >> salt-soluble fractions, although their reactivities were variable among patients and among the reaction conditions, indicating that the kind, the number and the epitope of allergens were variable among patients. To identify major allergens, alpha-, gamma- and omega-gliadin, and low-molecular-mass (LMM)- and high-molecular-mass (HMM)-glutenin genes were expressed in Escherichia coli by means of a pET vector. Recombinant gliadins and glutenins were partially purified on the basis of the solubilities of prolamin and glutelin. The partially purified recombinant proteins were reacted with the patients' sera. LMM glutenin containing many Gln-Gln-Gln-Pro-Pro motifs, which was identified to be IgE binding epitope [Tanabe, S., Arai, S., Yanagihara, Y., Mita, H., Takahashi, K. & Watanabe, M. (1996) Biochem. Biophys. Res. Commun. 219, 290-293], exhibited the highest reactivity. The next highest reactivities were observed on alpha-gliadin and gamma-gliadin, which had not been identified as allergens. PMID- 9738917 TI - Reassignment of the gene encoding the Escherichia coli hydrogenase 2 small subunit--identification of a soluble precursor of the small subunit in a hypB mutant. AB - An active tryptic fragment of hydrogenase 2 from Escherichia coli has been isolated from the periplasmic face of the cytoplasmic membrane, and the large and small subunits N-terminally sequenced. The large subunit is encoded by the hybC gene and shows no N-terminal processing, other than removal of the initiator methionine during its biosynthesis. Both N-terminal and the subsequent internal tryptic-fragment amino acid sequence indicate that the small subunit is neither encoded by hybA, a gene previously identified as encoding the small subunit [Menon et al. (1994) J. Bacteriol. 176, 4416-4423], nor any of the remaining genes in the hyb operon. Genome sequence analysis revealed the presence of an open reading frame which could potentially encode the peptide sequences of the proteolysed small subunit. The gene, designated hyb0, lies directly upstream of, and is separated by two nucleotides from, the start of the hybA gene. Hyb0, which shares an approximate 40% identity with other hydrogenase small subunit amino acid sequences, is synthesised with an N-terminal signal sequence containing a twin-arginine motif which is probably required for export of the enzyme. In the mature enzyme the small subunit is proteolytically cleaved after Ala37. Immunological analysis of strains overproducing either recombinant Hyb0 or HybA using antibodies specific for hydrogenase 2, readily identified Hyb0 as the small subunit. In a pleiotropic hypB mutant, which is unable to insert nickel into the active site, both the large and small subunits accumulate as unprocessed, soluble forms, consistent with the two subunits being assembled and processed in a coordinated manner during biosynthesis. PMID- 9738918 TI - Effect of molybdate and tungstate on the biosynthesis of CO dehydrogenase and the molybdopterin cytosine-dinucleotide-type of molybdenum cofactor in Hydrogenophaga pseudoflava. AB - The molybdenum-containing iron-sulfur flavoprotein CO dehydrogenase is expressed in a catalytically fully competent form during heterotrophic growth of the aerobic bacterium Hydrogenophaga pseudoflava with pyruvate plus CO. We have adopted these conditions for studying the effect of molybdate (Mo) and tungstate (W) on the biosynthesis of CO dehydrogenase and its molybdopterin (MPT) cytosine dinucleotide-(MCD)-type molybdenum cofactor. W was taken up by the Mo transport system and, therefore, interfered with Mo transport in an antagonistic way. Depletion of Mo from the growth medium as well as inclusion of excess W both resulted in the absence of intracellular Mo and led to the biosynthesis of CO dehydrogenase species of proper L2M2S2 subunit structure that carried the two 2Fe:2S type-I and type-II centers and two FAD molecules. EPR, ultraviolet/visible and CD spectroscopies established the full functionality of the cofactors. Due to the absence of the Mo-MCD cofactor, the enzyme species were catalytically inactive. Unexpectedly, the following cytidine nucleotides were present in inactive CO dehydrogenase: CDP, dCDP, CMP, dCMP, CTP or dCTP. The sum of cytidine nucleotides was two/mol enzyme. The binding specificities of inactive CO dehydrogenase for cytidine nucleotides (oxy > deoxy; diphosphate > monophosphate > triphosphate), and the absence of MPT suggest that, in active CO dehydrogenase, the cytidine diphosphate moiety of Mo-MCD provides the strongest interactions with the protein and determines the specificity for the type of nucleotide. In H. pseudoflava, the biosynthesis of MPT (identified as form A) was independent of Mo. Mo was, however, strictly required for the conversion of MPT to MCD (identified as form-A-CMP) as well as the insertion of Mo-MCD into CO dehydrogenase. These data support a model for the involvement of Mo in the biosynthesis of the Mo-MCD cofactor and of fully functional CO dehydrogenase in which the synthesis and insertion of Mo-MCD require Mo, and protein synthesis including integration of the FeS-centers and FAD are independent of Mo. PMID- 9738919 TI - Thrombin-induced reversal of astrocyte stellation is mediated by activation of protein kinase C beta-1. AB - Exogenous or endogenous injuries of the central nervous system trigger astrogliosis characterized by proliferation of astrocytes and changes in their morphology from stellate to flat polygonal. Astrocytes in culture are very sensitive to thrombin, a serine protease, which through its proteolytically activated receptor (PAR-1) induces proliferation and morphological changes comparable to astrogliosis. Evaluation of the thrombin signal-transduction pathway in the reversal of astrocyte stellation might help to understand astrogliosis. For this purpose, primary cultured murine cortical astrocytes were treated with H7, a protein-kinase inhibitor, and thrombin, which resulted in an inhibition of stellation reversal. Treatments with phorbol 12-myristate 13 acetate (PMA), a protein kinase C (PKC) activator, mimicked the action of thrombin. Subsequently, direct assay of astrocyte PKC activity after thrombin or PMA treatment demonstrated involvement of PKC in thrombin signaling associated with shape change. Western blotting showed that PKC isoform beta-1 was involved in this pathway, while PKC alpha was only weakly activated and PKC beta-2 was not activated by thrombin. PKC beta-1 translocation was also elicited by a thrombin receptor active peptide (SFLLRN), demonstrating the involvement of PAR-1 in this process. PKC delta and epsilon were located constitutively in the membrane fraction in stellate astrocytes. Isoforms gamma, eta, theta, and zeta were absent from astrocytes. These results suggest that astrogliosis in vitro might be regulated by modulating the activity of thrombin, PAR-1, or specific PKC isoforms. PMID- 9738920 TI - A novel aspect of calpain activation. AB - Calpain, a Ca2+-dependent biomodulator, alters the properties of substrate proteins by cleaving them at a limited number of specific sites. Recent studies of the structure-function relationship of calpain and X-ray analysis of its Ca2+ binding domain have revealed hitherto unknown features of the regulation of calpain activity. A novel dissociation/autolysis mechanism for the activation of calpain at the membrane is proposed, which incorporates recent findings from structure-function studies of calpain, and its implications are discussed. PMID- 9738921 TI - HIV: from molecular recognition to tissue pathogenesis. AB - Dramatic progress has been made recently in identifying both viral and cellular molecules responsible for binding and fusion of HIV-1 to target cells. In vivo, HIV-1 infection is transmitted by viruses that recognize chemokine receptor CCR5, while viruses isolated at later stages of HIV disease often recognize another chemokine receptor, CXCR4. It is still not understood how this molecular tropism of HIV-1 is translated into the virus' ability to compromise normal cell functions, which results in impairment of lymphoid tissue and causes AIDS. Here, we discuss how the new molecular findings might relate to HIV pathogenesis in cells and tissues. PMID- 9738922 TI - Photoaffinity labeling as an approach to study supramolecular nucleoprotein complexes. AB - The modern approaches for studying the detailed structure of nucleoprotein complexes involved in replication and transcription, based on the use of nucleic acids with photoreactive groups incorporated into definite positions of polynucleotide chain, are considered. Methods of preparation of photoreactive nucleic acids of this type are presented. Their use for positioning of RNA polymerase III and transcription factors as well as of the main participants of the replication machinery at the respective templates is described. A survey of the data concerning the amino acid residues modified in the course of photoaffinity labeling of proteins is also presented and some complications are discussed. PMID- 9738924 TI - Expression of matrix Gla protein (MGP) in an in vitro model of vascular calcification. AB - To investigate the role of matrix Gla protein (MGP), which can bind mineral ions through gamma-carboxylated glutamic acid residues, in vascular calcification, we examined the expression of MGP in an in vitro calcification model by using bovine vascular smooth muscle cells (BVSMC). The expression of MGP mRNA was decreased during BVSMC calcification and its levels were inversely correlated with the quantities of BVSMC calcification. MGP mRNA expression was restored to the level of uncalcified control by inhibiting BVSMC calcification with bisphosphonates. These data suggest that the expression of MGP gene is modulated in the development of vascular calcification. PMID- 9738923 TI - Down-regulation of negative acute-phase response genes by hypotonic stress in HepG2 hepatoma cells. AB - An increased hepatocellular hydration state (HS) that can be induced by hypotonic stress or a high glutamine uptake modulates the transcription of given genes in liver. This could be important in the acute phase (AP) of a systemic inflammation where both HS and glutamine uptake transiently increase in liver. In HepG2 hepatoma cells cultured in conditions of hypotonic stress or a high extracellular glutamine availability, a specifically decreased expression of two human mRNAs, namely those of alphal-microglobulin/bikunin precursor (AMBP) and alpha2-HS glycoprotein, that are also down-regulated in liver by AP, could be seen. A functional analysis of the AMBP promoter indicated that this hypotonic stress induced down-regulation takes place at a transcriptional level. In these experiments, the mRNA level and transcription of the glyceraldehyde-3-phosphate dehydrogenase gene that are known to be unmodified in AP did not exhibit any change. Given that hypotonic stress also upregulates the transcription of a liver gene that is also upregulated in AP [Meisse et al. (1998) FEBS Lett. 422, 3463481, the AP-associated increase in hepatocellular HS now appears to participate in the transcriptional control of both sets of genes that are up- or down-regulated in AP. PMID- 9738925 TI - Interleukin-2 stimulates a late increase in phosphatidic acid production in the absence of phospholipase D activation. AB - The signal transduction pathways involving phospholipid metabolism during T-cell proliferation remain partly undefined. Herein we show that interleukin-2 caused a late (> 12 h) rise in the intracellular phosphatidic acid content of CTLL-2 cells which was a consequence of the activation of the enzyme diacylglycerol kinase. No activation of phospholipase D was observed at similar times. Incubation of the cells with a recognized diacylglycerol kinase a isoform inhibitor, R59499, prior to interleukin-2 stimulation was able to block cell cycle entry, diacyglycerol kinase activation and phosphatidic acid accumulation. In contrast, when R59499 was added 3 h after interleukin-2, few or no observable effects on the above three parameters were noticed. These results suggest that the early signaling employed by IL-2 involving the alpha isoform of diacylglycerol kinase is sufficient to control the late increase in phosphatidic acid and that phosphatidic acid is a mitogenic agent in T-cells. PMID- 9738926 TI - The SHBG-like region of protein S is crucial for factor V-dependent APC-cofactor function. AB - Activated protein C (APC) regulates blood coagulation by degrading factor Va (FVa) and factor VIIIa (FVIIIa). Protein S is a cofactor to APC in the FVa degradation, whereas FVIIIa degradation is potentiated by the synergistic APC cofactor activity of protein S and factor V (FV). To elucidate the importance of the sex-hormone-binding globulin (SHBG)-like region in protein S for expression of anticoagulant activity, a recombinant protein S/Gas6 chimera was constructed. It comprised the amino-terminal half of protein S and the SHBG-like region of Gas6, a structurally similar protein having no known anticoagulant properties. The protein S/Gas6 chimera expressed 40-50%, APC-cofactor activity in plasma as compared to wild-type protein S. In the degradation of FVa by APC, the protein S/Gas6 chimera was only slightly less efficient than wild-type protein S. In contrast, the protein S/Gas6 chimera expressed no FV-dependent APC-cofactor activity in a FVIIIa-degradation system. This demonstrates the SHBG-like region to be important for expression of APC-cofactor activity of protein S and suggests that the SHBG-like region of protein S interacts with FV during the APC-mediated inactivation of FVIIIa. PMID- 9738927 TI - A different isoform of the transport protein mutated in the glycogen storage disease 1b is expressed in brain. AB - There are differences in the kinetic properties of the liver and brain microsomal glucose-6-phosphate transport systems suggesting the possibility of tissue specific isoforms. The availability of a human liver cDNA sequence which is mutated in patients with deficiencies of liver microsomal glucose-6-phosphate transport (glycogen storage disease 1b) made it possible to determine if a brain isoform exists. Northern blots of liver and brain RNA revealed that the mRNA of the brain form is slightly longer than the liver one. Isolation and sequencing of the respective human brain cDNA revealed that the brain protein has an additional 22 amino acid sequence. PMID- 9738928 TI - Brain-specific gene expression by immortalized microglial cell-mediated gene transfer in the mammalian brain. AB - The intra-arterial injection of immortalized microglia transfected with the lacZ gene, resulted in the expression of beta-galactosidase in the rat brain at 48 h and the activity of -galactosidase was detected for up to 3 weeks post-injection. More than 30-fold higher activity of beta-galactosidase was detected in the brain than in the liver, lung or spleen at 48 h post-injection. This method allows us to easily deliver the gene of interest to the brain without influencing other organs. Our brain-targeting gene delivery system can facilitate gene therapy of several brain disorders, including brain tumor, metabolic disorders, and degenerative disorders, as well as investigation into the roles of particular genes in brain function and development. PMID- 9738929 TI - C-terminal domain of beta-1,3-glucanase H in Bacillus circulans IAM1165 has a role in binding to insoluble beta-1,3-glucan. AB - The deduced amino acid sequences of 72-kDa beta-1,3-glucanase from Bacillus circulans WL-12 (GIcA) and 91-kDa enzyme from B. circulans IAM1165 (BglH) are highly homologous, except that the latter has an additional long C-terminal region composed of 192 amino acid residues. Two mutant enzymes (BgIH deprived of the C-terminal region and GIcA with the C-terminal region added) were constructed. The enzymes possessing the C-terminal region bound more abundantly to pachyman (insoluble beta-1,3-glucan) and A.spergillus oryzae cell wall than those not possessing the region. This indicates that the C-terminal region participated in binding of the enzymes to insoluble beta-1,3-glucan. PMID- 9738930 TI - Effect of green and black tea supplementation on lipids, lipid oxidation and fibrinogen in the hamster: mechanisms for the epidemiological benefits of tea drinking. AB - There is considerable epidemiological evidence that tea drinking lowers the risk of heart disease. However, the mechanism by which tea can be protective is unknown. Hamsters were fed a normal or high cholesterol diet for 2 weeks and drank green or black tea ad libitum. The plasma lipid profile was significantly improved by both teas compared to controls. Also in vivo lipid oxidation as measured by plasma lipid peroxides and LDL+VLDL oxidizability were significantly decreased by the teas. In the normal fed tea groups fibrinogen was decreased but not in the high cholesterol groups. Green tea was significantly more effective than the black tea. These results show in the hamster model that black and green tea improve the risk factors for heart disease by both hypolipemic and antioxidant mechanisms and possibly a fibrinolytic effect. PMID- 9738931 TI - Inhibitors of the proteasome block the myogenic differentiation of rat L6 myoblasts. AB - Myogenesis is characterized by membrane fusion and accumulation of muscle specific proteins. We have previously shown that nitric oxide acts as a messenger for membrane fusion. Here we show that inhibitors of the proteasome, such as lactacystin, reversibly block both the fusion of L6 myoblasts and the accumulation of muscle specific proteins, such as myosin heavy chain (MHC). The inhibitors also reversibly prevented the induction of the NF-kappaB activity, which is required for the expression of nitric oxide synthase (NOS). Moreover, the inhibition of the NF-kappaB activity occurred in parallel with that of the NOS activity upon treatment with increasing concentrations of lactacystin. While pyrrolidine dithiocarbamate, an inhibitor of NF-kappaB, blocked both membrane fusion and accumulation of MHC, N(G)-monomethyl-L-arginine, a specific inhibitor of NOS, inhibited only the fusion. These results suggest that the proteasome plays an essential role in the regulation of myogenic differentiation through the activation of NF-kappaB and that the target of NF-kappaB for the expression of muscle specific proteins is distinct from that for myoblast fusion. PMID- 9738932 TI - Specific cleavage of gamma catenin by caspases during apoptosis. AB - Caspase-mediated proteolysis of cytoskeletal proteins during apoptosis appears to be commonplace. Enlarging on previous studies we have shown here that gamma catenin, like beta catenin, was degraded during cisplatin-induced apoptosis, initially giving a major product of 75 kDa. This truncated protein could be co immunoprecipitated with alpha catenin. Addition of caspase inhibitors to cells in the presence of cisplatin appreciably reduced the proteolysis of gamma catenin as well as the level of apoptosis. Only limited degradation of alpha catenin was observed even at very late times when over 90% of cells in the culture were apoptotic. Immunohistochemical staining showed that during apoptosis there was a relocation of alpha, beta, and gamma catenin from the periphery of the cell to the cytoplasm, at the same time as other morphological changes commonly associated with apoptosis occurred. Interestingly, the changes in localisation of the catenins preceded proteolysis by several hours. In the presence of cisplatin and caspase inhibitor no change in distribution of catenins was observed, suggesting that re-localisation requires caspase activity but not necessarily directed against beta and gamma catenins. PMID- 9738933 TI - XAS characterization of the active sites of novel intradiol ring-cleaving dioxygenases: hydroxyquinol and chlorocatechol dioxygenases. AB - The intradiol cleaving dioxygenases hydroxyquinol 1,2-dioxygenase (HQI,20) from Nocardiodes simplex 3E, chlorocatechol 1,2-dioxygenase (CIC1,20) from Rhodococcus erythropolis ICP, and their anaerobic substrate adducts (hydroxyquinol-HQ1,20 and 4-chlorocatechol-CIC1,20) have been characterized through X-ray absorption spectroscopy. In both enzymes the iron(III) is pentacoordinated and the distance distribution inside the Fe(III) first coordination shell is close to that already found in the extensively characterized protocatechuate 3,4-dioxygenase. The coordination number and the bond lengths are not significantly affected by the substrate binding. Therefore it is confirmed that the displacement of a protein donor upon substrate binding has to be considered a general step valid for all intradiol dioxygenases. PMID- 9738934 TI - Identification of a novel human homolog of the Drosophila dlg, P-dlg, specifically expressed in the gland tissues and interacting with p55. AB - We have identified a novel human homolog of the Drosophila dlg tumor suppressor gene, termed P-dlg, which has been mapped at chromosome 10q23. Unlike other human dlg homologs, P-dlg is expressed in placenta and various gland tissues but not in brain. The P-dlg protein is localized at the plasma membrane and cytoplasm, and it is expressed in the gland epithelial cells in normal prostate tissue but not in prostate cancer cell lines. Furthermore, we identified interaction between P dlg and p55 palmitoylated membrane protein by yeast two-hybrid screening. These findings suggest that P-dlg forms a complex with p55 at the plasma membrane and plays roles in maintaining the structure of epithelial cells and transmitting extracellular signals to the membrane and cytoskeleton, which may negatively regulate cell proliferation. PMID- 9738935 TI - Evidence for a leukotriene A4 hydrolase in Xenopus laevis skin exudate. AB - Leukotriene A4 hydrolase is a cytosolic metalloenzyme of the arachidonic acid biosynthetic pathway responsible for leukotriene A4 conversion into leukotriene B4. In addition to its epoxide hydrolase properties, this enzyme exhibits an aminopeptidase activity which was used as an assay to monitor the purification of a novel form of leukotriene A4 hydrolase from Xenopus laevis skin exudate. This 70 kDa, secreted, form of leukotriene A4 hydrolase was identified by immunochemical cross-reactivity with anti-human leukotriene A4 hydrolase antibodies and by its capacity to convert leukotriene A4 into leukotriene B4. Moreover this enzyme produced a second metabolite which could be the leukotriene B4 isomer 5S,12R-dihydroxy-6,10-trans-8,14-4-cis-eicosatetraenoic acid, previously shown by Stromberg et al. (Eur.J. Biochem. 238 (1996) 599-605) to be formed by incubation of the leukotriene A4 with amphibian tissue extracts. Partial amino acid sequencing of peptides generated by endolysin C fragmentation of the purified enzyme confirmed the presence, in X. laevis skin secretions, of a related but distinct form of leukotriene A4 hydrolase which is likely to be responsible for the production of these eicosanoid metabolites of leukotriene A4. PMID- 9738936 TI - Direct association of presenilin-1 with beta-catenin. AB - Families bearing mutations in the presenilin-1 (PSI) gene develop Alzheimer's disease (AD). However, the mechanism through which PS1 causes AD is unclear. The co-immunoprecipitation with PS1 in transfected COS-7 cells indicates that PSI directly interacts with endogenous beta-catenin, and the interaction requires residues 322450 of PSI and 445-676 of beta-catenin. Both proteins are co localized in the endoplasmic reticulum. Over-expression of PS1 reduces the level of cytoplasmic beta-catenin, and inhibits beta-catenin-T cell factor-regulated transcription. These results indicate that PSI plays a role as inhibitor of the beta-catenin signal, which may be connected with the AD dysfunction. PMID- 9738937 TI - Synthesis of amidrazones using an engineered papain nitrile hydratase. AB - To demonstrate the usefulness of an engineered papain nitrile hydratase as a biocatalyst, a peptide amidrazone was prepared by incubation of the nitrile MeOCO Phe-Alanitrile with the Gln19Glu papain mutant in the presence of salicylic hydrazide as a nucleophile. The amidrazone results from nucleophilic attack by salicylic hydrazide at the imino carbon of the thioimidate adduct formed between the enzyme and the peptide nitrile substrate. Compared to wild-type enzyme, the engineered nitrile hydratase causes a better than 4000-fold increase in the rate of amidrazone formation and yields a product of much higher purity. The advantages over other nitrile-hydrolyzing enzymes and current limitations of the papain nitrile hydratase are discussed. PMID- 9738938 TI - Anchoring antibodies to membranes using a diphtheria toxin T domain-ZZ fusion protein as a pH sensitive membrane anchor. AB - We have constructed a fusion protein, T-ZZ, in which the IgG-Fc binding protein ZZ was fused to the C-terminus of the diphtheria toxin transmembrane domain (T domain). While soluble at neutral pH, T-ZZ retained the capacity of the T domain to bind to phospholipid membranes at acidic pH. Once anchored to the membrane, the ZZ part of the protein was capable of binding mouse monoclonal or rabbit polyclonal IgG. Our results show that the T-ZZ protein can function as a pH sensitive membrane anchor for the linkage of IgG to the membrane of lipid vesicles, adherent and non-adherent cells. PMID- 9738939 TI - A residue in the middle of the M2-M3 loop of the beta4 subunit specifically affects gating of neuronal nicotinic receptors. AB - An aspartate residue in the M2-M3 loop of neuronal nicotinic receptor alpha7 subunits is a major determinant of the channel functional response. This residue is conserved in most beta4 subunits, e.g. human and rat, but not in others, e.g. bovine. We have used these differences to examine the mechanism by which this residue alters the functional properties of alpha3beta4 receptors. Having ruled out an effect on the macroscopic binding ability of the agonist, the level of receptor expression, or the single channel conductance, the results suggest that receptors lacking that residue have a deficient coupling between binding and gating. PMID- 9738940 TI - Intrinsic uncoupling of cytochrome c oxidase may cause the maternally inherited mitochondrial diseases MELAS and LHON. AB - Mutations in the human mtDNA gene encoding subunit III of cytochrome c oxidase (CO) have been reported to cause MELAS and LHON. Poracoccus denitrificans cells expressing substitutions homologous to these MELAS- and LHON-causing mutations had lower growth yield than wild type cells and lower efficiency of proton pumping by CO (e.g. lower H+/e ratio and lower deltapsi), but had similar CO activity. These results indicate that both substitutions (F263L > A212T) cause intrinsic uncoupling, which may be the direct cause of the diseases. These results also suggest that subunit III is involved in proton pumping. PMID- 9738941 TI - Computational analyses and annotations of the Arabidopsis peroxidase gene family. AB - Classical heme-containing plant peroxidases have been ascribed a wide variety of functional roles related to development, defense, lignification, and hormonal signaling. More than 40 peroxidase genes are now known in Arabidopsis thaliana for which functional association is complicated by a general lack of peroxidase substrate specificity. Computational analysis was performed on 30 near full length Arabidopsis peroxidase cDNAs for annotation of start codons and signal peptide cleavage sites. A compositional analysis revealed that 23 of the 30 peroxidase cDNAs have 5' untranslated regions containing 40-71% adenine, a rare feature observed also in cDNAs which predominantly encode stress-induced proteins, and which may indicate translational regulation. PMID- 9738942 TI - Use of the phage display technique for detection of epitopes recognized by polyclonal rabbit gliadin antibodies. AB - A random phage heptapeptide library was screened with rabbit antibodies against wheat flour proteins comprising gliadins and a small amount of low molecular weight glutenins (gli/glu). Gli/glu antibodies isolated from the sera selected different consensus sequences (CS). All CS contained tri- to pentapeptide stretches homologous to gli/glu sequences (proposed epitopes). In alpha- and gamma-type gliadins, these sequences are clustered in the N-terminal region recently suspected to be toxic for humans with celiac disease. Peptides with CS were synthesized and checked for reactivity. Only immune and no control rabbit sera reacted with synthetic peptides. One of eight human sera containing gliadin antibodies was reactive as well (4/8 peptides) but control sera were negative. Thus the phage display technique is useful for epitope screening of polyclonal antibodies even in the case of a group of homologous but diverse antigens. PMID- 9738943 TI - Evolution of new protein function: recombinational enhancer Fis originated by horizontal gene transfer from the transcriptional regulator NtrC. AB - New protein function is thought to evolve mostly by gene duplication and divergence. Here we present phylogenetic evidence that the multifunctional protein Fis of the gamma proteobacterial species derived from the COOH-terminal domain of an ancestral alpha proteobacterial NtrC transcriptional regulatory protein. All of the known enterobacterial fis genes are preceded by an open reading frame, named yhdG, that is highly similar to nifR3, a gene that forms an operon with ntrC in several alpha proteobacterial species. Thus, we propose that yhdG and fis were acquired by a lineage ancestral to the gamma proteobacteria in a single horizontal gene transfer event, and later diverged to their present functions. PMID- 9738944 TI - Molecular cloning of two novel types of peptidylarginine deiminase cDNAs from retinoic acid-treated culture of a newborn rat keratinocyte cell line. AB - Peptidylarginine deiminases (PADs) are a group of enzymes which convert protein arginine residues to citrulline residues. Using rat muscle PAD cDNA as a probe, we obtained two novel cDNAs, PAD-R11 and PAD-R4, from immortalized rat keratinocytes treated with all-trans retinoic acid. Comparison of the deduced amino acid sequences with those of muscle and hair follicle enzymes showed high conservation in the C-terminal region. Recombinant proteins encoded by both PAD R11 and PAD-R4 showed the enzyme activities. That of PAD-R11 showed a characteristic feature of the enzyme found in the epidermis. PMID- 9738945 TI - Structure and organization of the cardiotoxin genes in Naja naja sputatrix. AB - We report the genomic structure, organization and the presence of multiple isoforms of the gene encoding cardiotoxins (CTX) of Naja naja sputatrix. The cardiotoxin gene consists of six CTX isoforms, each (2.2 kb) having three exons and two introns. Two possible transcription initiation sites as well as consensus TATA boxes and transcription factor binding motifs, AP-2, NFIL-6/C/EBP, NF-kappaB and PuF have been identified in the 5'-region of the gene. The CTX gene isoforms show nucleotide variations at specific segments in exon 2 and exon 3, which correspond to the functional domains in the three-finger loop structure of the cardiotoxin molecule. The diverse functions of cardiotoxins together with our findings suggest that the cardiotoxin gene isoforms may have evolved under adaptive pressure through a positive Darwinian selection process. PMID- 9738946 TI - Nitric oxide induces and inhibits apoptosis through different pathways. AB - Physiological levels of nitric oxide (NO) regulate vascular tone and protect the microvasculature from injury whereas excessive NO may be harmful. The present study explored the effects of NO on human endothelial cell apoptosis. We found that the NO donor S-nitroso-N-acetylpenicillamine (SNAP) inhibited TNFalpha induced endothelial apoptosis and that this was mediated partly through the cGMP pathway. In contrast, high SNAP concentration induced endothelial apoptosis via cGMP-independent pathways and the cGMP pathway protected against NO-induced apoptosis. These findings demonstrate that low NO concentrations contribute to human endothelial cell survival, whereas higher NO concentrations are pathological and promote destruction of endothelial cells. PMID- 9738947 TI - Acylation of Streptomyces type II polyketide synthase acyl carrier proteins. AB - Acyl derivatives of type II PKS ACPs are required for in vitro studies of polyketide biosynthesis. The presence of an exposed cysteine residue prevented specific chemical acylation of the phosphopantetheine thiol of the actinorhodin PKS holo ACP. Acylation studies were further complicated by intramolecular disulphide formation between cysteine 17 and the phosphopantetheine. The presence of this intramolecular disulphide was confirmed by tryptic digestion of the ACP followed by ESMS analysis of the fragments. An act Cys17Ser ACP was engineered by site-directed mutagenesis. S-Acyl adducts of act C17S, oxytetracycline and griseusin holo ACPs were rapidly formed by reaction with hexanoyl, 5-ketohexanoyl and protected acetoacetyl imidazolides. Comparisons with type 11 FAS ACPs were made. PMID- 9738948 TI - The endogenous cannabinoid anandamide potentiates interleukin-6 production by astrocytes infected with Theiler's murine encephalomyelitis virus by a receptor mediated pathway. AB - Theiler's murine encephalomyelitis virus (TMEV) infection of a susceptible strain of mice results in virus persistence in the brain and chronic primary immune mediated demyelination, which resembles multiple sclerosis. Recent attention has focused on the anti-inflammatory and immunosuppressive properties of interleukin 6, a pleiotropic cytokine involved in the regulation of immunological responses, acute phase protein production and hematopoiesis. Anandamide (arachidonoyl ethanolamine) is a natural brain constituent that binds a specific brain cannabinoid receptor. In this study we investigated whether anandamide can modify interleukin-6 production by primary cultures of murine brain cortical astrocytes infected with TMEV. Astrocytes from susceptible (SJL/J) and resistant (BALB/c) strains of mice infected with TMEV (10(5)PFU/well) increased IL-6 release over a period of 24 h. Anandamide caused an enhancement of the release of IL-6 by TMEV infected astrocytes in a concentration-dependent manner (1-25 microM). Treatment of TMEV-infected astrocytes with 10 microM arachidonyl trifluoromethyl ketone, a potent inhibitor of the amidase that degrades anandamide, was found to potentiate the effects of anandamide on IL-6 release. A novel and selective cannabinoid receptor antagonist, SR 141617A, blocked the enhancing effects of anandamide on IL-6 release by TMEV-infected astrocytes, suggesting a cannabinoid receptor mediated pathway. The physiological implications of these results are unknown, but may be related to the hypothesis of the protective effects of cannabinoids on neurological disorders like multiple sclerosis. PMID- 9738949 TI - cDNA cloning and characterization of mouse nifS-like protein, m-Nfs1: mitochondrial localization of eukaryotic NifS-like proteins. AB - We have isolated a mouse cDNA which shows significant sequence similarity to the yeast nifS-like gene (y-NFS1), and termed it m-Nfs1. The deduced protein sequence (459 amino acids long) has several characteristic features common to those of bacterial NifS proteins, but distinct from them by its amino-terminal extension which contains a typical mitochondrial targeting presequence. m-Nfs1 was found to be a soluble 47-kDa protein in the matrix fraction of mouse liver mitochondria. The m-Nfs1 gene was ubiquitously expressed in most tissues, suggesting its housekeeping function in vivo. We also found that the gamma-NFS1 protein was localized in the mitochondrial matrix in yeast cells. These results suggest that both eukaryotic NifS-like proteins may play some roles in mitochondrial functions. PMID- 9738950 TI - cDNA cloning and inducible expression of human multidrug resistance associated protein 3 (MRP3). AB - Previously, we cloned rat MRP3 as a candidate for an inducible transporter for the biliary excretion of organic anions [Hirohashi et al. (1998) Mol. Pharmacol. 53, 1068-10751. In the present study, we cloned human MRP3 (1527 amino acids) from Caco-2 cells. Human MRP3 is predominantly expressed in liver, small intestine and colon; hepatic expression of MRP3 was observed in humans but not in normal rats. In HepG2 cells, the expression of MRP3 was induced by phenobarbital. These results suggest that MRP3 may act as an inducible transporter in the biliary and intestinal excretion of organic anions. PMID- 9738951 TI - Conformational analysis of the interdomain linker of the central homology region of chloroplast initiation factor IF3 supports a structural model of two compact domains connected by a flexible tether. AB - A peptide corresponding to the interdomain linker of chloroplast IF3 has been synthesized and its structure studied by NMR and CD as a function of temperature and pH. At low temperature and neutral pH the apparent helical content is 25%. pH and ionic strength dependent CD studies demonstrate that sidechain-sidechain interactions stabilize the structure observed at low temperature. The helicity decreases with temperature and above 25 degrees C the peptide is less than 15% helical. These results indicate that the peptide has little intrinsic tendency to form helical structure at physiologically relevant temperatures and strongly suggests that the linker region is flexible in intact chloroplast IF3. PMID- 9738952 TI - Properties of the Ca2+ influx reveal the duality of events underlying the activation by vanadate and fluoride of the Gardos effect in human red blood cells. AB - The properties of the 45Ca2+ influx by human red blood cells (RBC) induced by NaVO3 or NaF were compared. The NaVO3-induced 45Ca2+ influx was slower and less extensive than that induced by NaF. Both processes were saturable with Ca2+. Substitution of Na+ by K+ inhibited the 45Ca2+ influx induced by NaVO3 but stimulated that by NaF. The NaVO3-induced Ca2+ influx was sensitive to nifedipine (IC50 = 50 mol/l), Cu2+ (IC50=9 mol/l), DTNB (5,5'-dithiobis-(dinitrobenzoic acid)) (IC50 = 12 mol/l) (maximal inhibition 16%, 18%, and 28%, respectively, if NaF was used as inducer). On the other hand, tetrodotoxin (TTX) and cyclosporin A inhibited only the NaF-induced 45Ca2+ influx (IC50 = 21 mol/l and 28 mol/l, respectively). Pig RBC, known not to display the NaVO3-induced Ca2+ influx, exhibited Ca2+ influx induced by NaF. The results show that NaVO3 activates the Ca2+ influx via a pathway homologous to the L-type Ca2+ channel while the NaF induced Ca2+ influx is mediated via the TTX-sensitive Na+ channel in the presence of NaF with possible participation of calcineurin or cyclophilin. Thus, the Gardos effect induced by NaVO3 and NaF represents two phenomena activated by different mechanisms present in the cryptic state in the RBC membrane. PMID- 9738953 TI - Lysophosphatidylcholine upregulates CD40 ligand expression in newly activated human CD4+ T cells. AB - Lysophosphatidylcholine (lyso-PC) accumulates in tissues undergoing inflammation and atherosclerosis, where an infiltration of T cells is also seen. We found that lyso-PC increased IFN-gamma production and CD40L expression in CD4+ T cells stimulated with anti-CD3 Ab and recombinant CD80 molecules, whereas lyso-PC did not affect IL-2 and IL-4 production. These results suggest that lyso-PC, in combination with other stimuli, may regulate CD4+ T cell functions to propagate local inflammatory reactions and also imply a novel role played by a modified lipid in the selection of Th1/Th2 immune response as well as in the T cell mediated pathogenesis in atherosclerosis. PMID- 9738954 TI - Association of extracellular-superoxide dismutase phenotype with the endothelial constitutive nitric oxide synthase polymorphism. AB - The distribution of extracellular-superoxide dismutase (EC-SOD) levels in healthy Australian subjects was consistent with two distinct phenotypes in which the smaller group of subjects (3.3%) had 15-fold higher levels. The EC-SOD levels in individuals homozygous for endothelial constitutive nitric oxide synthase 4a (ecNOS4a), a rare allele for ecNOS repeat polymorphism at intron 4, were significantly lower than those in ecNOS4A/a and ecNOS4AIA subjects. Furthermore, NO levels were negatively correlated with the EC-SOD levels in common EC-SOD phenotype subjects. Whilst the mechanism remains speculative, it is possible that there is a significant interaction between EC-SOD and ecNOS, or that common factor(s), either genetic or environmental, influence both of them. PMID- 9738955 TI - Membrane fusion plays an important role in gene transfection mediated by cationic liposomes. AB - By confocal laser scanning microscopy (CLSM) we have studied the membrane fusion between cationic liposomes and the endosome membranes involved in gene transfection mediated by cationic liposomes. Antisense oligonucleotides were transferred by cationic liposomes with a cationic cholesterol derivative, cholesteryl-3beta-carboxyamidoethylenedimethylamine (I). Cationic liposomes were made by a mixture of the derivative I and DOPE. The intracellular distribution of fluorescein-conjugated antisense oligonucleotides (phosphorothioate) was studied by CLSM. The images showed that the antisense oligonucleotides were preferentially transferred into the nucleus of target cells (NIH3T3, COS-7 and HeLa cells) by the liposomes with derivative I. However, their transfection was completely blocked by nigericin which was able to dissipate the pH gradient across the endosome membranes, although the liposome/DNA complex was found in the cytoplasm of the target cells. This was quite in contrast with the fluorescence images of the target cells treated with wortmannin, an inhibitor of endocytosis. The results suggest that at least two steps are effective for gene transfection mediated by the cationic liposomes with cationic cholesterol derivatives. One is the endocytosis of the liposome/DNA complex into the target cells and the other is the removal of antisense oligonucleotides (plasmid DNAs) from the complex in the endosomes. The latter step was preferentially preceded by the membrane fusion between the cationic liposomes and the endosome membranes at around pH 5.0. PMID- 9738956 TI - DNA binding of NF-Y: the effect of HMGI proteins depends upon the CCAAT box. AB - NF-Y is a conserved sequence-specific transcription factor binding to CCAAT boxes. The chromatin-associated HMGI proteins influence promoter activities through positive and negative effects on binding of transcription factors. It was previously shown that HMGI(Y) synergizes the binding of NF-Y to the alpha2 collagen CCAAT box [Currie, R.A. (1997) J. Biol Chem. 272, 30880-30888]. Using recombinant proteins, we confirm that at low concentrations of NF-Y, HMGI(Y) acts synergistically on the alpha2-collagen CCAAT and we extend this observation to HMGI and HMGI-C. However, enhancement of DNA binding to gamma-globin, alpha globin and MHC class II Ea CCAAT boxes was not observed. At high concentrations, HMGI proteins inhibit binding to alpha2-collagen and to gamma-globin, but not to high affinity Ea or a-globin CCAAT. In none of our experiments did we see a ternary complex between NF-Y, HMGI(Y) and DNA. In protein competition experiments, NF-Y affinity was at least two orders of magnitude higher, even in the context of the suboptimal gamma-globin CCAAT. Our data prove that HMGI proteins have complex positive and negative effects on NF binding to some, but not to all CCAAT boxes, suggesting that this phenomenon is dictated by the sequences flanking the pentanucleotide rather than direct protein-protein interactions. PMID- 9738957 TI - The mouse gene encoding the peroxisomal membrane protein 1-like protein (PXMP1 L): cDNA cloning, genomic organization and comparative expression studies. AB - PXMPI-L (synonyms: PMP69, P70R) is a peroxisomal protein that belongs to the ABC transporter superfamily. Its closest homolog is the peroxisomal membrane protein 1 (PMP70). We have cloned the mouse PXMP1-L gene. It encodes a 606 amino acid protein. In contrast to the human and the rat, mouse PXMP1-L is predominantly expressed in the liver. The mouse PXMP1-L gene consists of 19 exons and spans 21 kb of genomic sequence. No obvious peroxisome proliferator response element has been found in 1.1 kb of the putative promoter region. No coordination of constitutive or fenofibrate-induced expression of PXMP1-L with other peroxisomal ABC transporters was observed so that an obligate exclusive heterodimer formation is not likely to occur. The data presented will be particularly useful for the generation of a mouse model defective in PXMP1-L in order to elucidate the yet unknown function of this protein. PMID- 9738959 TI - N-glycoprotein biosynthesis in plants: recent developments and future trends. AB - N-glycosylation is a major modification of proteins in plant cells. This process starts in the endoplasmic reticulum by the co-translational transfer of a precursor oligosaccharide to specific asparagine residues of the nascent polypeptide chain. Processing of this oligosaccharide into high-mannose-type, paucimannosidic-type, hybrid-type or complex-type N-glycans occurs in the secretory pathway as the glycoprotein moves from the endoplasmic reticulum to its final destination. At the end of their maturation, some plant N-glycans have typical structures that differ from those found in their mammalian counterpart by the absence of sialic acid and the presence of beta(1,2)-xylose and alpha( 1,3) fucose residues. Glycosidases and glycosyltransferases that respectively catalyse the stepwise trimming and addition of sugar residues are generally considered as working in a co-ordinated and highly ordered fashion to form mature N-glycans. On the basis of this assembly line concept, fast progress is currently made by using N-linked glycan structures as milestones of the intracellular transport of proteins along the plant secretory pathway. Further developments of this approach will need to more precisely define the topological distribution of glycosyltransferases within a plant Golgi stack. In contrast with their acknowledged role in the targeting of lysosomal hydrolases in mammalian cells, N glycans have no specific function in the transport of glycoproteins into the plant vacuole. However, the presence of N-glycans, regardless of their structures, is necessary for an efficient secretion of plant glycoproteins. In the biotechnology field, transgenic plants are rapidly emerging as an important system for the production of recombinant glycoproteins intended for therapeutic purposes, which is a strong motivation to speed up research in plant glycobiology. In this regard, the potential and limits of plant cells as a factory for the production of mammalian glycoproteins will be illustrated. PMID- 9738958 TI - The endoplasmic reticulum of plant cells and its role in protein maturation and biogenesis of oil bodies. AB - The endoplasmic reticulum (ER) is the port of entry of proteins into the endomembrane system, and it is also involved in lipid biosynthesis and storage. This organelle contains a number of soluble and membrane-associated enzymes and molecular chaperones, which assist the folding and maturation of proteins and the deposition of lipid storage compounds. The regulation of translocation of proteins into the ER and their subsequent maturation within the organelle have been studied in detail in mammalian and yeast cells, and more recently also in plants. These studies showed that in general the functions of the ER in protein synthesis and maturation have been highly conserved between the different organisms. Yet, the ER of plants possesses some additional functions not found in mammalian and yeast cells. This compartment is involved in cell to cell communication via the plasmodesmata, and, in specialized cells, it serves as a storage site for proteins. The plant ER is also equipped with enzymes and structural proteins which are involved in the process of oil body biogenesis and lipid storage. In this review we discuss the components of the plant ER and their function in protein maturation and biogenesis of oil bodies. Due to the large number of cited papers, we were not able to cite all individual references and in many cases we refer the readers to reviews and references therein. We apologize to the authors whose references are not cited. PMID- 9738962 TI - Compartment-specific accumulation of recombinant immunoglobulins in plant cells: an essential tool for antibody production and immunomodulation of physiological functions and pathogen activity. AB - Expression and stability of immunoglobulins in transgenic plants have been investigated and optimized by accumulation in different cellular compartments as cytosol, apoplastic space and endoplasmic reticulum (ER) as will be discussed in this review. In several cases described the highest accumulation of complete active antibodies was achieved by targeting into the apoplastic space. High-level expression of active recombinant single-chain Fv antibodies (scFv's) was obtained by retention of these proteins in the lumen of the endoplasmic reticulum. This has been shown for leaves and seeds of transgenic tobacco as well as for potato tubers. Transgenic tobacco seeds, potato tubers and tobacco leaves can facilitate stable storage of scFv's accumulated in the ER over an extended (seeds, tubers) or a short (leaves) period of time. The expression of specific scFv's in different plant species, plant organs and cellular compartments offers the possibility of blocking regulatory factors or pathogens specifically. Examples are scFv's expressed in the cytosol and the apoplastic space of transgenic plant cells modulating the infection process of plant viruses and a cytosolically expressed scFv that influenced the activity of phytochrome A protein. The immunomodulation approach has been shown to be also applicable for investigating the action of the phyto-hormone abscisic acid (ABA). High-level accumulation of specific anti-ABA scFv's in the ER of all leaf cells has been used to block the influence of ABA on the stomatal functions. Seed-specific expression of high amounts of anti-ABA-scFv's at a defined time of seed-development induced a developmental switch from seed ripening to vegetative growth. It has been demonstrated that ER retention is essential for the accumulation of sufficient scFv to bind high concentrations of ABA in the transgenic seeds. PMID- 9738961 TI - Deposition of storage proteins. AB - Plants store amino acids for longer periods in the form of specific storage proteins. These are deposited in seeds, in root and shoot tubers, in the wood and bark parenchyma of trees and in other vegetative organs. Storage proteins are protected against uncontrolled premature degradation by several mechanisms. The major one is to deposit the storage proteins into specialized membrane-bounded storage organelles, called protein bodies (PB). In the endosperm cells of maize and rice prolamins are sequestered into PBs which are derived from the endoplasmic reticulum (ER). Globulins, the typical storage proteins of dicotyledonous plants, and prolamins of some cereals are transported from the ER through the Golgi apparatus and then into protein storage vacuoles (PSV) which later become transformed into PBs. Sorting and targeting of storage proteins begins during their biosynthesis on membrane-bound polysomes where an N-terminal signal peptide mediates their segregation into the lumen of the ER. After cleavage of the signal peptide, the polypeptides are glycosylated and folded with the aid of chaperones. While still in the ER, disulfide bridges are formed which stabilize the structure and several polypeptides are joined to form an oligomer which has the proper conformation to be either deposited in ER-derived PB or to be further transferred to the PSV. At the trans-Golgi cisternae transport vesicles are sequestered which carry the storage proteins to the PSV. Several storage proteins are also processed after arriving in the PSVs in order to generate a conformation that is capable of final deposition. Some storage protein precursors have short N- or C-terminal targeting sequences which are detached after arrival in the PSV. Others have been shown to have internal sequence regions which could act as targeting information. In some cases positive targeting information is known to mediate sorting into the PSV whereas in other cases aggregation and membrane association seem to be major sorting mechanisms. PMID- 9738963 TI - Exocytosis in plants. AB - Exocytosis is the final event in the secretory pathway and requires the fusion of the secretory vesicle membrane with the plasma membrane. It results in the release to the outside of vesicle cargo from the cell interior and also the delivery of vesicle membrane and proteins to the plasma membrane. An electrophysiological assay that measures changes in membrane capacitance has recently been used to monitor exocytosis in plants. This complements information derived from earlier light and electron microscope studies, and allows both transient and irreversible fusion of single exocytotic vesicles to be followed with high resolution in protoplasts. It also provides a tool to investigate bulk exocytotic activity in single protoplasts under the influence of cytoplasmic modulators. This research highlights the role of intracellular Ca2+, GTP and pressure in the control of exocytosis in plants. In parallel to these functional studies, plant proteins with the potential to regulate exocytosis are being identified by molecular analysis. In this review we describe these electrophysiological and molecular advances, and emphasise the need for parallel biochemical work to provide a complete picture of the mechanisms controlling vesicle fusion at the plasma membrane of plant cells. PMID- 9738964 TI - Sorting of proteins to vacuoles in plant cells. AB - An individual plant cell may contain at least two functionally and structurally distinct types of vacuoles: protein storage vacuoles and lytic vacuoles. Presumably a cell that stores proteins in vacuoles must maintain these separate compartments to prevent exposure of the storage proteins to an acidified environment with active hydrolytic enzymes where they would be degraded. Thus, the organization of the secretory pathway in plant cells, which includes the vacuoles, has a fascinating complexity not anticipated from the extensive genetic and biochemical studies of the secretory pathway in yeast. Plant cells must generate the membranes to form two separate types of tonoplast, maintain them as separate organelles, and direct soluble proteins from the secretory flow specifically to one or the other via separate vesicular pathways. Individual soluble and membrane proteins must be recognized and sorted into one or the other pathway by distinct, specific mechanisms. Here we review the emerging picture of how separate plant vacuoles are organized structurally and how proteins are recognized and sorted to each type. PMID- 9738960 TI - The molecular characterization of transport vesicles. AB - Secretion, endocytosis and transport to the lytic compartment are fundamental, highly coordinated features of the eukaryotic cell. These intracellular transport processes are facilitated by vesicles, many of which are small (100 nm or less in diameter) and 'coated' on their cytoplasmic surface. Research into the structure of the coat proteins and how they interact with the components of the vesicle membrane to ensure the selective packaging of the cargo molecules and their correct targeting, has been quite extensive in mammalian and yeast cell biology. By contrast, our knowledge of the corresponding types of transport vesicles in plant cells is limited. Nevertheless, the available data indicate that a considerable homology between plant and non-plant coat polypeptides exists, and it is also suggestive of a certain similarity in the mechanisms underlying targeting in all eukaryotes. In this article we shall concentrate on three major types of transport vesicles: clathrin-coated vesicles, COP-coated vesicles, and 'dense' vesicles, the latter of which are responsible for the transport of vacuolar storage proteins in maturing legume cotyledons. For each we will summarize the current literature on animal and yeast cells, and then present the relevant data derived from work on plant cells. In addition, we briefly review the evidence in support of the 'SNARE' hypothesis, which explains how vesicles find and fuse with their target membrane. PMID- 9738965 TI - The nuclear pore complex. AB - The nuclear pore complex is the largest supramolecular complex that assembles in the eukaryotic cell. This structure is highly dynamic and must disassemble prior to mitosis and reassemble after the event. The directed movement of macromolecules into and out of the nucleus occurs through the nuclear pore complex, a potentially regulatory point for translocation. Using biochemical and genetic approaches, several nuclear pore complex proteins from yeast and vertebrates have been well characterized. Although very little is known about plant nuclear pore proteins, research is providing new information that indicates that plant nuclear pore complexes may have some unique features. PMID- 9738967 TI - Protein translocation into and across the chloroplastic envelope membranes. AB - Post-translational protein import into chloroplasts follows a common route characterised by the need for nucleoside-triphosphates at various steps and two distinct protein import machineries at the outer and inner envelope membrane, respectively. Several subunits of these complexes have been elucidated. In contrast, protein translocation into the chloroplastic outer envelope uses distinct and various but poorly characterised insertion pathways. A topological framework for single-membrane spanning proteins of the chloroplastic outer envelope is presented. PMID- 9738968 TI - Multiple pathways for the targeting of thylakoid proteins in chloroplasts. AB - The assembly of the photosynthetic apparatus requires the import of numerous cytosolically synthesised proteins and their correct targeting into or across the thylakoid membrane. Biochemical and genetic studies have revealed the operation of several targeting pathways for these proteins, some of which are used for thylakoid lumen proteins whereas others are utilised by membrane proteins. Some pathways can be traced back to the prokaryotic ancestors of chloroplasts but at least one pathway appears to have arisen in response to the transfer of genes from the organelle to the nucleus. In this article we review recent findings in this field that point to the operation of a mechanistically unique protein translocase in both plastids and bacteria, and we discuss emerging data that reconcile the remarkable variety of targeting pathways with the natures of the substrate precursor proteins. PMID- 9738966 TI - The surprising complexity of peroxisome biogenesis. AB - Peroxisomes are small organelles with a single boundary membrane. All of their matrix proteins are nuclear-encoded, synthesized on free ribosomes in the cytosol, and post-translationally transported into the organelle. This may sound familiar, but in fact, peroxisome biogenesis is proving to be surprisingly unique. First, there are several classes of plant peroxisomes, each specialized for a different metabolic function and sequestering specific matrix enzymes. Second, although the mechanisms of peroxisomal protein import are conserved between the classes, multiple pathways of protein targeting and translocation have been defined. At least two different types of targeting signals direct proteins to the peroxisome matrix. The most common peroxisomal targeting signal is a tripeptide limited to the carboxyl terminus of the protein. Some peroxisomal proteins possess an amino-terminal signal which may be cleaved after import. Each targeting signal interacts with a different cytosolic receptor; other cytosolic factors or chaperones may also form a complex with the peroxisomal protein before it docks on the membrane. Peroxisomes have the unusual capacity to import proteins that are fully folded or assembled into oligomers. Although at least 20 proteins (mostly peroxins) are required for peroxisome biogenesis, the role of only a few of these have been determined. Future efforts will be directed towards an understanding of how these proteins interact and contribute to the complex process of protein import into peroxisomes. PMID- 9738970 TI - Protein import into cyanelles and complex chloroplasts. AB - Higher-plant, green and red algal chloroplasts are surrounded by a double membrane envelope. The glaucocystophyte plastid (cyanelle) has retained a prokaryotic cell wall between the two envelope membranes. The complex chloroplasts of Euglena and dinoflagellates are surrounded by three membranes while the complex chloroplasts of chlorarachniophytes, cryptomonads, brown algae, diatoms and other chromophytes, are surrounded by 4 membranes. The peptidoglycan layer of the cyanelle envelope and the additional membranes of complex chloroplasts provide barriers to chloroplast protein import not present in the simpler double membrane chloroplast envelope. Analysis of presequence structure and in vitro import experiments indicate that proteins are imported directly from the cytoplasm across the two envelope membranes and peptidoglycan layer into cyanelles. Protein import into complex chloroplasts is however fundamentally different. Analysis of presequence structure and in vitro import into microsomal membranes has shown that translocation into the ER is the first step for protein import into complex chloroplasts enclosed by three or four membranes. In vivo pulse chase experiments and immunoelectronmicroscopy have shown that in Euglena, proteins are transported from the ER to the Golgi apparatus prior to import across the three chloroplast membranes. Ultrastructural studies and the presence of ribosomes on the outermost of the four envelope membranes suggests protein import into 4 membrane-bounded complex chloroplasts is directly from the ER like outermost membrane into the chloroplast. The fundamental difference in import mechanisms, posttranslational direct chloroplast import or co-translational translocation into the ER prior to chloroplast import, appears to reflect the evolutionary origin of the different chloroplast types. Chloroplasts with a two membrane envelope are thought to have evolved through the primary endosymbiotic association between a eukaryotic host and a photosynthetic prokaryote while complex chloroplasts are believed to have evolved through a secondary endosymbiotic association between a heterotrophic or possibly phototrophic eukaryotic host and a photosynthetic eukaryote. PMID- 9738969 TI - The role of lipids in plastid protein transport. AB - The elaborate compartmentalization of plant cells requires multiple mechanisms of protein targeting and trafficking. In addition to the organelles found in all eukaryotes, the plant cell contains a semi-autonomous organelle, the plastid. The plastid is not only the most active site of protein transport in the cell, but with its three membranes and three aqueous compartments, it also represents the most topologically complex organelle in the cell. The chloroplast contains both a protein import system in the envelope and multiple protein export systems in the thylakoid. Although significant advances have identified several proteinaceous components of the protein import and export apparatuses, the lipids found within plastid membranes are also emerging as important players in the targeting, insertion, and assembly of proteins in plastid membranes. The apparent affinity of chloroplast transit peptides for chloroplast lipids and the tendency for unsaturated MGDG to adopt a hexagonal II phase organization are discussed as possible mechanisms for initiating the binding and/or translocation of precursors to plastid membranes. Other important roles for lipids in plastid biogenesis are addressed, including the spontaneous insertion of proteins into the outer envelope and thylakoid, the role of cubic lipid structures in targeting and assembly of proteins to the prolamellar body, and the repair process of D1 after photoinhibition. The current progress in the identification of the genes and their associated mutations in galactolipid biosynthesis is discussed. Finally, the potential role of plastid-derived tubules in facilitating macromolecular transport between plastids and other cellular organelles is discussed. PMID- 9738971 TI - Two birds with one stone: genes that encode products targeted to two or more compartments. AB - Eukaryotic cells are divided into multiple membrane-bound compartments, all of which contain proteins. A large subset of these proteins perform functions that are required in more than one compartment. Although in most cases proteins carrying out the same function in different compartments are encoded by different genes, this is not always true. Numerous examples have now been found where a single gene encodes proteins (or RNAs) found in two (or more) cell organelles or membrane systems. Some particularly clear examples come from protein synthesis itself: plant cells contain three protein-synthesizing compartments, the cytosol, the mitochondrial matrix and the plastid stroma. All three compartments thus require tRNAs and aminoacyl-tRNA synthetases. Some mitochondrial tRNAs and their aminoacyl-tRNA synthetases are identical to their cytosolic counterparts and they are encoded by the same genes. Similarly, some mitochondrial and plastid aminoacyl-tRNA synthetases are encoded by the same nuclear genes. The various ways in which differentially targeted products can be generated from single genes is discussed. PMID- 9738974 TI - Lung cancer patients have increased 8-hydroxydeoxyguanosine levels in peripheral lung tissue DNA. AB - The 8-hydroxydeoxyguanosine (8-OH-dG) levels in the peripheral parts of human lung tissues were compared between lung cancer patients (n=70) and non-cancer patient controls (n=15). An increased level of 8-OH-dG was observed in the lung cancer group, in both the adenocarcinoma and non-adenocarcinoma (mainly squamous cell carcinoma) groups, as compared to the non-cancer control group. This result suggests that reactive oxygen species are partly involved in the induction of lung carcinomas (both adenocarcinoma and non-adenocarcinoma). PMID- 9738972 TI - Intercellular protein trafficking through plasmodesmata. AB - During plant morphogenesis, groups of cells differentiate to form specialized tissues possessing distinct structures and functions. Cell specialization is a result of specific gene expression at the individual cell level. Coordination of differential gene expression among cells requires that cells communicate with one another. Plasmodesmata provide a cytoplasmic pathway for direct intercellular communication. Recent discoveries that macromolecules such as transcription factors, viral proteins, and plant defense-related proteins can traffic through plasmodesmata suggest that intercellular protein trafficking is potentially an important means to regulate plant developmental processes, physiological functions, plant-pathogen interactions, and plant defense reactions. Thus, elucidating the specific functions and mechanisms of intercellular protein trafficking has broad implications in understanding how a plant develops and functions at the molecular level. This review is to provide an update on this rapidly developing area of plant biology, with emphasis on the discussion of possible mechanisms underlying intercellular protein trafficking. PMID- 9738975 TI - Significant correlation of nitric oxide synthase activity and p53 gene mutation in stage I lung adenocarcinoma. AB - Nitric oxide (NO) and its derivatives can directly cause DNA damage and mutation in vitro and may play a role in the multistage carcinogenic process. It has been reported that NO induces mutation in the p53 tumor suppressor gene; we therefore analyzed the relationship between NO synthase (NOS) activity and p53 gene status in early-stage lung adenocarcinoma. Surgical samples were classified into two categories: 14 lung adenocarcinomas with high NOS activity (>25 pmol/min/g tissue, category A), and 16 with low NOS activity (<25 pmol/min/g tissue, category B). A yeast functional assay for p53 mutations disclosed a red colony that corresponded to a mutation in the p53 gene in 8 cases (57.1%) in category A and 3 cases (18.8%) in category B, the frequency being significantly higher in the former (P<0.05). A p53 DNA sequence analysis revealed that 5 of the 8 p53 mutation-positive samples in category A had a G:C-to-T:A transversion, which is reported to be a major target of NO. The mechanism of carcinogenesis of adenocarcinoma is not fully understood, but these results suggest that an excess of endogenously formed NO may induce a p53 gene mutation containing mainly G:C-to T:A transversion in the early stage of lung adenocarcinoma. Our results suggest that NO has potential mutagenic and carcinogenic activity, and may play important roles in human lung adenocarcinoma. PMID- 9738977 TI - Expression of the TCL1 gene at 14q32 in B-cell malignancies but not in adult T cell leukemia. AB - The TCL1 gene was recently cloned as a candidate target within the 14q32.1 breakpoint cluster region observed in T-cell malignancies. We examined the TCL1 gene expression in 21 patients with adult T-cell leukemia (ATL) and 5 cell lines, because ATL is reported to have frequent chromosome 14 band q32 aberrations. However, 20 of the ATL patients and all 5 cell lines lacked any TCL1 expression on northern blot analysis, and TCL1 transcripts were only very faintly detected in the remaining one patient. Expansion of our analysis to include other types of hematopoietic malignancies revealed strong expression of the TCL1 gene in almost all tumor cells of B-cell lineage except myelomas. However, no TCL1 signals were encountered in cells of T-cell or myeloid lineages. In normal human tissues TCL1 was found to be expressed in the spleen, lymph nodes and B-lymphocytes of peripheral blood. These results indicate that TCL1 is not a major target gene for ATL, but that it may play a role in B-cell differentiation and proliferation. PMID- 9738976 TI - Molecular analysis of Ewing's sarcoma: another fusion gene, EWS-E1AF, available for diagnosis. AB - Ewing's sarcoma, one of the most malignant tumors of children and young adults, expresses specific chimeric genes, e.g. EWS-FLI-1, EWS-ERG, EWS-ETV1 and EWS-FEV. In this paper, we extensively characterized a new fusion gene, EWS-EIAF by means of whole cDNA sequencing, RNA blot analysis, DNA blot analysis and chromosomal analysis, and showed it to be available for the diagnosis of Ewing's sarcoma and to participate in the oncogenesis of Ewing's sarcoma. Furthermore, we conducted a genetic analysis of Ewing family tumors in conjunction with immunohistochemical analysis and ultrastructural analysis. Our results demonstrate some limitations of both genetic analysis and histopathological analysis, and establish the relationship between neurogenic phenotypes and chimera genes. PMID- 9738973 TI - Mitochondrial protein import in plants. Signals, sorting, targeting, processing and regulation. AB - Mitochondrial biogenesis requires a coordinated expression of both the nuclear and the organellar genomes and specific intracellular protein trafficking, processing and assembly machinery. Most mitochondrial proteins are synthesised as precursor proteins containing an N-terminal extension which functions as a targeting signal, which is proteolytically cleaved off after import into mitochondria. We review our present knowledge on components and mechanisms involved in the mitochondrial protein import process in plants. This encompasses properties of targeting peptides, sorting of precursor proteins between mitochondria and chloroplasts, signal recognition, mechanism of translocation across the mitochondrial membranes and the role of cytosolic and organellar molecular chaperones in this process. The mitochondrial protein processing in plants is catalysed by the mitochondrial processing peptidase (MPP), which in contrast to other sources, is integrated into the bc1 complex of the respiratory chain. This is the most studied component of the plant import machinery characterised to date. What are the biochemical consequences of the integration of the MPP into an oligomeric protein complex and how are several hundred presequences of precursor proteins with no sequence similarities and no consensus for cleavage, specifically cleaved off by MPP? Finally we will address the emerging area of the control of protein import into mitochondria. PMID- 9738979 TI - Surgical significance of telomerase activity in noncancerous liver tissue from patients with hepatocellular carcinoma. AB - Telomerase activity has been detected in tissue from noncancerous liver of patients with chronic liver disease, but its functional significance remains to be elucidated. We therefore evaluated the telomerase activity in surgically obtained noncancerous liver tissue from 20 hepatocellular carcinoma (HCC) patients. Two samples of noncancerous liver tissue were obtained from each patient: one from the parenchyma adjacent to the HCC nodules of the resected specimen; the other from the parenchyma distant from the HCC nodules of the remnant liver. Telomerase activity was assayed by a non-radioisotope quantitative system based on "TRAP-eze." Five samples from the noncancerous liver tissue adjacent to the HCC nodules (25.0%) were telomerase-positive; all such cases showed high-grade malignant potential, such as intrahepatic metastasis and/or portal vascular invasion and infiltration of the fibrous capsule in the corresponding HCC nodules, and telomerase positivity showed neither a relationship with the histological activity index scores nor a correlation with liver function. Interestingly, no telomerase activity was detected in any of the 20 samples obtained from the parenchyma of the remnant liver. These results indicate that telomerase in noncancerous liver tissue is associated not with the hepatic condition accompanying HCC, but with the biological characteristics of the tumor itself, and may derive from infiltrating cancer cells. Determination of telomerase status may aid in designing more effective surgical procedures. PMID- 9738978 TI - Expression of G1-->S transition regulatory molecules in human urothelial cancer. AB - Growth of cancer cells is characterized by accelerated passage through the cell cycle, which is often caused by deregulation of the G1-->S transition. In this study the expression of G1-->S transition regulatory molecules was analyzed in 32 transitional cell carcinoma specimens and fifteen normal tissues obtained by cystectomy or nephroureterectomy of mainly locally advanced tumors, as well as six bladder cancer cell lines. Expression of mRNAs for cyclins D1 and D2 and cyclin-dependent kinases (CDK) 2 and 4 was investigated by quantitative reverse transcription-polymerase chain reaction. Overexpression of cyclin D1 compared to normal mucosa was observed in 3 tumors (9.4%), but in neither of the cell lines. All tumors with overexpression were moderately differentiated (G2) pT1 or pT2 tumors, and thus among the less advanced specimens. Cyclin D2 was not expressed in normal bladder mucosa or in tumors. The expression of CDK4 mRNA varied within the same range in mucosa, tumors, and cell lines. CDK2 mRNA expression varied more strongly and was diminished in individual tumors and in four cell lines. It is concluded that cyclin D1 overexpression can play an important role in the early stage of urothelial tumorigenesis, driving cell proliferation. Ectopic expression of cyclin D2 or amplification of CDK4 does not occur at a significant frequency in urothelial carcinomas. Different expression patterns of cyclin D1 and CDK2 indicate heterogeneity in the mechanisms of G1-->S transition deregulation in individual bladder tumors which may elicit differences in their biological and clinical behavior. PMID- 9738980 TI - Inhibitory effects of ginsenoside Rh2 on tumor growth in nude mice bearing human ovarian cancer cells. AB - Ginsenoside Rh2 (Rh2), isolated from an ethanol extract of the processed root of Panax ginseng CA Meyer, inhibits the growth of B16 melanoma cells. This study was designed to evaluate the ability of Rh2 to inhibit growth of human ovarian cancer cells (HRA) in vitro and in nude mouse. Rh2 inhibited proliferations of various established human ovarian cancer cell lines in a dose-dependent manner between 10 and 60 microM in vitro and induced apoptosis at around the IC50 dose. When HRA cells were inoculated s.c. into the right flank of nude mice, all mice formed a palpable tumor within 14 days. Although i.p. administration of Rh2 alone hardly inhibited the tumor growth, when Rh2 was combined with cis diamminedichloroplatinum(II) (CDDP) the tumor growth was significantly inhibited, compared to treatment with CDDP alone. When mice were treated p.o. with Rh2 daily (but not weekly), the tumor growth was significantly (P<0.01) inhibited, compared to CDDP treatment alone. When Rh2 was combined with CDDP, the degree of tumor growth retardation was not potentiated. The survival time was significantly (P<0.05) longer than that of medium alone-treated controls or the group treated with CDDP alone. Then, we examined whether p.o. administration of Rh2 has a dose dependent inhibitory effect on the tumor growth. I.p. and weekly administration of CDDP had more potent antitumor activity in the order of 1 mg/kg, 2 mg/kg and 4 mg/kg, whereas p.o. and daily administration of Rh, (0.4 to 1.6 mg/kg) not only had antitumor activity comparable to that of 4 mg/kg CDDP, but also resulted in a significant increase of the survival. Doses of Rh2 used in this study did not result in any adverse side-effects as confirmed by monitoring hematocrit values and body weight, unlike 4 mg/kg CDDP, which had severe side-effects. It is noteworthy that p.o. but not i.p. treatment with Rh2 resulted in induction of apoptotic cells in the tumor in addition to augmentation of the natural killer activity in spleen cells from tumor-hearing nude mice. Thus, particularly in view of the toxicity of CDDP, Rh2 alone would seem to warrant further evaluation for treatment of recurrent or refractory ovarian tumor. PMID- 9738981 TI - Fas and mutant estrogen receptor chimeric gene: a novel suicide vector for tamoxifen-inducible apoptosis. AB - Several cancer gene therapy strategies involve suicide genes to kill the neoplasm, or to regulate effector cells such as lymphocytes. We have developed an inducible apoptosis system with a Fas-estrogen receptor fusion protein (MfasER) for rapid elimination of transduced cells. In the present study, we further improved this molecular switch for estrogen-inducible apoptosis to overcome concerns with the wild-type estrogen receptor and its natural ligand, 17beta estradiol (E2). The ligand-binding domain of MfasER was replaced with that of a mutant estrogen receptor which is unable to bind estrogen yet retains affinity for a synthetic ligand, 4-hydroxytamoxifen (Tm). The resultant fusion protein (MfasTmR) and MfasER were expressed in L929 cells for examination of their ligand specificities. Tm induced apoptosis in MfasTmR-expressing cells (L929MfasTmR) at 10(-8) M or higher concentrations, but induced no apoptosis in MfasER-expressing cells (L929MfasER) at up to 10(-6) M. On the other hand, E2 induced apoptosis in L929MfasER at concentrations as low as 10(-10)-10(-9) M, while it did so partially in L929MfasTmR at concentrations greater than 10(-7) M. Thus, L929MfasTmR cells were highly susceptible to Tm, but refractory to E2, with 100 1,000 times more tolerance than L929MfasER. These results suggest that the MfasTmR/Tm system would induce apoptosis in the target cells more safely in vivo, working independently of endogenous estrogen. PMID- 9738982 TI - Growth inhibition of CD20-positive B lymphoma cell lines by IDEC-C2B8 anti-CD20 monoclonal antibody. AB - Treatment with IDEC-C2B8 (C2B8), the chimeric anti-CD20 antibody, was shown in a phase I-II study to be very effective for the treatment of low-grade B-cell lymphoma, in contrast to the results of most previous immunotherapies with monoclonal antibodies. In a study designed to elucidate the reason for this efficacy, two cell lines derived from lymphomas with BCL2 gene rearrangement (SU DHL-4 and SU-DHL-6) showed remarkable growth inhibition and cell-death, and two other cell lines derived from a diffuse lymphoma (RC-K8) and a mantle cell lymphoma (SP-49) showed moderate growth inhibition, but neither a CD20 weakly positive cell line (NALL-1) nor a negative cell line (MOLT-4) showed any growth inhibition. An examination of the intensity of cell-surface CD20 expression showed no correlation between intensity and degree of growth inhibition among the four cell lines showing growth inhibition. Morphological examination revealed condensed and fragmented nuclei and budding of the plasma membrane, both characteristic of apoptosis, with some cells in these cell lines showing growth inhibition by C2B8. Such apoptosis was also confirmed by flow cytometric analysis, suggesting that, at least in part, apoptosis plays a role in this growth inhibition. This growth-inhibitory mechanism may thus account for the effectiveness of C2B8 antibody therapy. PMID- 9738983 TI - Release of cytokines from human umbilical vein endothelial cells treated with platinum compounds in vitro. AB - Endothelial cells (EC) produce cytokines, such as interleukin (IL)-1, IL-6, IL-8 and granulocyte-macrophage colony-stimulating factor (GM-CSF). These cytokines have an important role in the proliferation and differentiation of hematopoietic progenitor cells. On the other hand, anticancer agents generally cause hematopoietic disorders. However, little is known about the effects of chemotherapeutic agents on the secretion of cytokines from EC. Therefore, we investigated if treatment with platinum compounds may stimulate EC to secrete cytokines. EC newly isolated from a human umbilical vein were exposed to cisplatin, carboplatin, or TRK-710 for 80 min, then the cells were washed and placed in fresh medium. The levels of cytokines in the fresh medium were measured by the ELISA method, the levels of intracellular hydrogen peroxide (H2O2) were measured by flow cytometry, and the rhodamine 123-stained live mitochondria of the EC were observed under a confocal laser microscope. Platinum compounds induced cytokine production in human EC: cisplatin most prominently induced the release of IL-1 and IL-6, and TRK-710 had the greatest ability to induce the release of GM-CSF. Intracellular H2O2 production and IL-8 release were transiently induced immediately after treatment with platinum compounds, leading to IL-1 release when H2O2 production was eliminated. These results may provide new insights into the hematological toxicity induced by anticancer agents and the role of IL-1 and IL-6 secreted from EC in this toxicity. PMID- 9738984 TI - Applicability of combination with tirapazamine in boron neutron capture therapy. AB - SCC VII tumor-bearing mice were continuously given 5-bromo-2'-deoxyuridine (BrdU) to label all proliferating cells. After injection of tirapazamine (TPZ), a bioreductive agent, combined with sodium borocaptate-10B (BSH) or dl-p boronophenylalanine-10B (BPA) administration, the tumors were irradiated with thermal neutrons, and then isolated and incubated with cytochalasin-B (a cytokinesis blocker). The micronucleus (MN) frequency in cells without BrdU labeling (quiescent (Q) cells) was determined by means of immunofluorescence staining for BrdU, and that for total cells was obtained from tumors not pretreated with BrdU. Even when no 10B-compound was administered, TPZ increased the MN frequency of tumor cells including Q cells, resulting in reduction of the difference in MN frequency between total and Q cells, mainly by increasing the MN frequency of Q cells. TPZ increased the MN frequency of Q cells when combined with BPA administration, but TPZ showed no apparent effect on each cell population when combined with BSH. Namely, TPZ reduced the difference in MN frequency between total and Q cells caused by 10B-compound administration, especially when BPA was administered. From the viewpoint of the overall cell killing effect in boron neutron capture therapy (BNCT), combination with TPZ appeared to be useful in BPA-BNCT, but not in BSH-BNCT. PMID- 9738985 TI - Antitumor immunity induction by intracellular hyperthermia using magnetite cationic liposomes. AB - Induction of antitumor immunity to T-9 rat glioma by intracellular hyperthermia using functional magnetic particles was investigated. Magnetite cationic liposomes (MCLs), which have a positive surface charge, were used as heating mediators for intracellular hyperthermia. Solid T-9 glioma tissues were formed subcutaneously on both femurs of female F344 rats, and MCLs were injected via a needle only into the left solid tumors (treatment side). The rats were then divided into two groups, which received no irradiation, or irradiation for 30 min given three times at 24-h intervals with an alternating magnetic field (118 kHz, 384 Oe). On the treatment side, the tumor tissue disappeared completely in many rats exposed to the magnetic field. The tumor tissue on the opposite side also disappeared completely, even though MCLs were not injected into the right solid tumors. To examine whether a long-lasting and tumor-specific immunity could be generated, the rats that had been cured by the hyperthermia treatment were rechallenged with T-9 cells 3 months later. After a period of transient growth, all tumors disappeared. Furthermore, immunocytochemical assay revealed that the immune response induced by the hyperthermia treatment was mediated by both CD8+ and CD4+ T cells and accompanied by a marked augmentation of tumor-selective cytotoxic T lymphocyte activity. These results suggest that our magnetic particles are potentially effective tools for hyperthermic treatment of solid tumors, because in addition to killing of the tumor cells by heat, a host immune response is induced. PMID- 9738986 TI - Fluorescence in situ hybridization analysis of 12;21 translocation in Japanese childhood acute lymphoblastic leukemia. AB - Fluorescence in situ hybridization (FISH) analysis was applied to detect t(12;21) using two yeast artificial chromosome probes and cosmid probes covering the TEL(ETV6) and the AML1 gene to clarify the incidence of abnormality of t(12;21) in Japanese childhood acute lymphoblastic leukemia (ALL). We detected seven TEL/AML1 fusion positive patients (9.5%), all of whom were diagnosed as B-lineage ALL, among 74 childhood ALL. On the other hand, no TEL/AML1 fusion positive patients were found among 37 adult ALL. The incidence among Japanese seemed to be lower than that among other nations. Of the seven patients with the TEL/AML1 fusion, five exhibited normal karyotype, one was t(8;12)(q11;p13), i(21q) and the remaining one exhibited a near-triploid karyotype in conventional G-banding. The FISH method clearly demonstrated that all patients with the TEL/AML1 fusion had subpopulations of leukemic cells with deletion of the normal TEL allele, which is significant for understanding the progression of leukemia with t(12;21). PMID- 9738987 TI - Perinatal infectious disease. Introduction. PMID- 9738988 TI - Bacterial vaginosis: association with adverse pregnancy outcome. AB - Bacterial vaginosis is the most common lower genital tract infection encountered among women of reproductive age. This condition can best be considered as a vaginal syndrome associated with an alteration of the normal vaginal flora rather than an infection specific to any one microorganism. Bacterial vaginosis is a clinical condition with a complex microbiology that is characterized by a reduced concentration of a normally abundant Lactobacillus species along with high concentrations of gram-negative and anaerobic bacteria, particularly, Gardnerella vaginalis and Mobiluncus, Bacteroides, Prevotella, and Mycoplasma species. The exact make up of the microorganisms and their relative concentration vary among women who have this condition. Although it was previously regarded as a harmless condition, recent work has linked bacterial vaginosis to numerous upper genital tract complications such as preterm labor and preterm delivery, preterm premature rupture of the membranes, chorioamnionitis, and postpartum endometritis. The findings from recent prospective randomized trials suggest that treatment of bacterial vaginosis in certain women who are at high risk for preterm delivery decreases the rate of preterm birth. PMID- 9738989 TI - Pathophysiology, diagnosis, and management of intraamniotic infection. AB - Intraamniotic infection (IAI) is a term used to describe a clinically diagnosed infection of the contents of the uterus. It is found most often after rupture of the membranes. The most useful diagnostic tests are physical examination, amniotic fluid glucose determination, and amniotic fluid Gram's stain. There is no clearly established means for the prevention of IAI, but cervical examinations and cervical manipulation can increase the risk, so caution with their use is still warranted. Treatment for this infection should be initiated when the diagnosis is made to provide the lowest risk of neonatal and maternal complications. Ampicillin or penicillin plus gentamicin are the most extensively tested antibiotics for treatment before delivery. Clindamycin or metronidazole should be added if a cesarean section is performed. As a general rule, antibiotics should be continued postpartum until the patient has been afebrile and asymptomatic for a minimum of 24 hours. Neonatal complications of IAI may be substantial especially for the premature fetus. Women with this infection have a greater risk for dysfunctional labor and cesarean section. PMID- 9738990 TI - Cytomegalovirus infection. AB - Cytomegalovirus (CMV) infection is the most common perinatal infection and may result in severe injury to the fetus. Forty percent to 50% of infants delivered to mothers with primary CMV will have congenital infections. Of these, 5% to 18% will be overtly symptomatic at birth. The mortality rate in these children is almost 30%; approximately 80% of the survivors have severe neurological morbidity. The majority of congenitally infected infants will be asymptomatic at birth; 10% to 15% of these children subsequently have sequelae such as visual and auditory defects. If recurrent or reactivated CMV infection develops during pregnancy, the risk of serious fetal injury is very low. Similarly, neonatal infection acquired during delivery or from breast feeding also poses minimal risk to the child. Because antimicrobial therapy and immunoprophylaxis for CMV infection are unsatisfactory, pregnant women must be educated about preventive measures. PMID- 9738991 TI - Group B streptococcal infections. AB - Group B streptococcal infection is the most common cause of neonatal sepsis and is responsible for significant neonatal morbidity and mortality. Group B streptococcus is also the causative agent in 50,000 maternal infections per year. Approximately 30% of women have asymptomatic group B streptococcal colonization at some time during pregnancy, but the neonatal attack rate is only about 2 per 1,000 deliveries. Maternal and neonatal risk factors contribute to the rates of vertical transmission and symptomatic neonatal disease. Options that have been investigated for prevention of neonatal group B streptococcal disease include identification of at-risk pregnancies as well as antenatal, intrapartum, and neonatal treatment. The intrapartum treatment of women at risk for vertical transmission of group B streptococcus to their neonates unequivocally has been shown to decrease the rate of neonatal colonization. Practitioners should implement one of two strategies that incorporate intrapartum prophylaxis for prevention of perinatal group B disease. PMID- 9738992 TI - Hepatitis in pregnancy. AB - Currently, six distinct types of hepatitis virus have been identified: A, B, C, D, E, and G. Hepatitis A virus infection does not cause a chronic carrier state, and perinatal transmission is extremely uncommon. Hepatitis B can be transmitted perinatally, but immunization of the newborn with hepatitis B immune globulin and hepatitis B vaccine markedly reduces the risk of neonatal infection. Hepatitis D virus is dependent on coinfection with the hepatitis B virus for replication. Immunoprophylaxis against hepatitis B also is effective against hepatitis D. Hepatitis C virus is primarily transmitted by the parenteral route and is particularly likely to cause chronic liver disease. Perinatal transmission of hepatitis C principally occurs in women who have high titers of HCV-RNA or who are coinfected with human immunodeficiency virus. At this time, no immunoprophylaxis for hepatitis C is available. Hepatitis G, a recently described organism, is related to hepatitis C. Its clinical significance remains undetermined. Hepatitis E is transmitted in a manner similar to hepatitis A. Perinatal transmission is unusual, but maternal disease is often severe. PMID- 9738993 TI - Herpes simplex virus. AB - Herpes simplex virus (HSV) infection is prevalent worldwide. Herpes labialis, caused predominantly by HSV-1, and herpes vulvovaginitis, caused predominantly by HSV-2, may result in significant morbidity and mortality for infected neonates exposed during delivery. The diagnosis of HSV infection is made by serological testing, viral culture, or polymerase chain reaction. Women with primary herpes vulvovaginitis exhibit a painful vesicular rash which is self-limited but may be followed by multiple recurrences. Women at greatest risk to transmit HSV to their neonates are those who experience their first episode of HSV during the latter stage of pregnancy. If infected, their neonates may have localized skin, eye and mucosal lesions, invasive central nervous system infection, or disseminated disease. Because of the potentially devastating outcome for a baby infected with HSV, pregnant women with active HSV lesions at delivery should be offered a cesarean section. Still, many neonates who are infected with HSV are born to women with asymptomatic HSV shedding. Therefore, prevention of HSV during pregnancy is exceedingly important. PMID- 9738994 TI - Human immunodeficiency virus infection in pregnancy. AB - The pace at which our knowledge and treatment of the human immunodeficiency virus (HIV) has advanced has been staggering. A disease that was unknown two decades ago, that was untreatable only a decade ago, and whose rate of mother-to-child transmission was immutable just 5 years ago, is now readily diagnosed, treated with increasing effectiveness, and blocked from transmission in the large majority of cases. None of these advances can be provided to patients unless their physicians actively screen patients and, for those identified as HIV infected, assure them of access to the latest therapies. This article is a primer for those obstetricians who would engage in such efforts. The data that form the basis of therapy are provided as well as clinical guidelines for the care of the pregnant woman infected with HIV. PMID- 9738995 TI - Parvovirus B19 infections in pregnancy. AB - Parvovirus B19 is the viral agent that causes the childhood exanthum erythema infectiosum, or fifth disease. Approximately 50% of pregnant women are seropositive for this agent and thus immune to primary infection. However, acute infection may develop in seronegative pregnant women exposed to B19. Acute B19 infections during pregnancy have been associated with miscarriage and hydrops fetalis. This latter condition is amenable to fetal therapy via intrauterine transfusion. PMID- 9738996 TI - Rubella and rubeola. AB - Rubella and rubeola are common viral exanthems that may affect women of reproductive age. Effective vaccination programs have greatly decreased their incidence. Although Rubella is a relatively innocuous illness for the nonpregnant patient, transplacental fetal infection with rubella can result in significant and crippling fetal malformations and handicap. Because some women of reproductive age are not appropriately immunized, rubella is still a threat. The practitioner needs to be vigilant in assuring vaccination of susceptible individuals when seen for routine health maintenance. Additionally, at times the obstetrician will be challenged with the evaluation and care of a susceptible pregnant patient who is exposed to rubella. In contrast, rubeola (measles) infection during pregnancy has not been associated with congenital malformations. Affected mothers, however, experience a higher incidence of spontaneous abortions and premature delivery and are themselves at risk for serious complications such as pneumonia and encephalitis. PMID- 9738997 TI - Syphilis. AB - Syphilis was first recognized as a distinct syndrome in Europe in the fifteenth century. Despite knowledge of congenital infection for more than 450 years and the existence of adequate therapy for 55 years, congenital infection remains a problem for the practicing clinician. Syphilis is caused by Treponema pallidum. Infection may be transmitted horizontally by sexual contact and vertically as a result of hematogenous dissemination across the placenta. Syphilis is classified as primary, secondary, latent, and tertiary. The diagnosis may be established by darkfield examination of clinical lesions and by serological assays. The drug of choice for syphilis is penicillin. This agent is the only antibiotic of proven value for the treatment of congenital syphilis. Accordingly, infected pregnant women who are allergic to beta-lactam antibiotics must be desensitized and then treated with penicillin. PMID- 9738998 TI - Toxoplasmosis. AB - Toxoplasmosis is caused by the protozoan organism, Toxoplasma gondii. Infection with this organism primarily results from contact with infected cats and from ingestion of improperly cooked meat. Most adults with toxoplasmosis are asymptomatic. When symptoms are present, they typically resemble a mononucleosis or flulike illness. The diagnosis of toxoplasmosis in the pregnant adult is best made using serological techniques to detect IgM antibody and to document significant changes in the IgG antibody titer. Congenital toxoplasmosis usually occurs as a result of primary maternal infection. The most useful tests for confirmation of fetal infection are ultrasound examination, cordocentesis for detection of IgM-specific antibody, and amniocentesis for detection of toxoplasma DNA in amniotic fluid. Congenital toxoplasmosis can be treated with reasonable success by administration of antibiotics (spiramycin, sulfadiazine, and pyrimethamine) to the mother. In an effort to prevent acquisition of infection, pregnant women should be counseled to avoid contact with cat litter and improperly cooked beef, pork, or lamb. PMID- 9739000 TI - Medicine and science in the life of Luigi Galvani (1737-1798). AB - Together with its companion paper, dealing with the contribution of Luigi Galvani to the history of electrophysiology, this article provides a biographical sketch of the scientist of Bologna in the occasion of the bicentenary of his death. Studies on Galvani have focused mainly on his "discovery" of animal electricity, and on the controversy with Alessandro Volta. Much less is known about Galvani's life and activity as a teacher, physician, and researcher in the fields of comparative anatomy, physiology, and chemistry of life. Yet, a balanced assessment of the significance and the role of Galvani's research in the history of science will be possible only after a historical reconstruction of his entire activity. This should take into account aspects of Galvani's life that have been little studied up to now: Galvani's scientific background, the scientific context in which his interest for muscular physiology arose, the interplay between his activity as a researcher and as a physician, the origin and characteristics of his experimental approach to biological studies, and the development of his experimental research in the crucial period culminating in his electrophysiological explanation of muscular motion. The present article aims at offering a contribution in this direction. PMID- 9738999 TI - Varicella in pregnancy. AB - Varicella-zoster virus may cause serious infection, particularly pneumonia, in adult women. Women of child-bearing age should be questioned about immunity to varicella preconceptually, and offered serological testing, and VARIVAX vaccine if indicated. All pregnant patients should be questioned about immunity to varicella during their first prenatal appointment. Susceptible patients should be counseled to avoid contact with individuals who have chickenpox. If exposure occurs, VZIG should be administered within 96 hours in an attempt to prevent maternal infection. Varicella embryopathy may occur as a result of maternal infection particularly in the first half of pregnancy with an incidence of 1% to 2%. Varicella of the newborn is a life-threatening illness that may occur when a newborn is delivered within 5 days of the onset of maternal illness or after postdelivery exposure to varicella. Susceptible neonates should receive VZIG. Acyclovir is active against the varicella-zoster virus, and treatment is indicated in seriously ill adults and neonates. PMID- 9739001 TI - Animal electricity and the birth of electrophysiology: the legacy of Luigi Galvani. AB - Preceded by a companion paper on Galvani's life, this article is written on the occasion of the bicentenary of the death of Luigi Galvani. From his studies on the effects of electricity on frogs, the scientist of Bologna derived the hypothesis that animal tissues are endowed with an intrinsic electricity that is involved in fundamental physiological processes such as nerve conduction and muscle contraction. Galvani's work swept away from life sciences mysterious fluids and elusive entities like "animal spirits" and led to the foundation of a new science, electrophysiology. Two centuries of research work have demonstrated how insightful was Galvani's conception of animal electricity. Nevertheless, the scholar of Bologna is still largely misrepresented in the history of science, because the importance of his researches seems to be limited to the fact that they opened the paths to the studies of the physicist Alessandro Volta, which culminated in 1800 with the invention of the electric battery. Volta strongly opposed Galvani's theories on animal electricity. The matter of the scientific controversy between Galvani and Volta is examined here in the light of two centuries of electrophysiological studies leading to the modern understanding of electrical excitability in nerve and muscle. By surveying the work of scientists such as Nobili, Matteucci, du Bois-Reymond, von Helmholtz, Bernstein, Hermann, Lucas, Adrian, Hodgkin, Huxley, and Katz, the real matter of the debate raised by Galvani's discoveries is here reconsidered. In addition, a revolutionary phase of the 18th century science that opened the way for the development of modern neurosciences is reevaluated. PMID- 9739002 TI - The organization of chemically characterized afferents to the perivascular neuronal groups of the hypothalamic magnocellular neurosecretory system in the rat. AB - The hypothalamic magnocellular neurosecretory system consists of the paraventricular nucleus and supraoptic nucleus and a number of accessory nuclei. There is evidence that each of the accessory nuclei has a preferential source of afferents. Two of the accessory nuclei, namely the nucleus circularis (NC) and the lateral hypothalamic perivascular nucleus (LHPN), are particularly interesting due to their very close relationship with the blood vessels. The NC is composed of small dense clusters of neurons in the medial anterior hypothalamus. The groups of lateral hypothalamic neurons gathering around vascular branches are collectively called the LHPN. Their close topographical relationship with the blood vessels may indicate that the latter may serve as a source of input to these nuclei. As a part of the effort to investigate this issue, the present study examined in these two nuclei the distribution pattern of terminal-like elements containing 11 transmitters/modulators. Only a few, if any, terminal-like elements of the transmitters/modulators studied could be found distributed in the NC proper, although its immediate vicinity could be densely innervated. On the contrary, the LHPN proper was often densely innervated by fibers expressing the examined markers. These terminal patterns were found to be quite different from those of the paraventricular and supraoptic nuclei. The present findings further substantiate the notion of a functional differentiation among the subnuclei of the magnocellular neurosecretory system. The significance of the relationship of these two perivascular nuclei with the blood vessels is discussed. PMID- 9739003 TI - Effect of 5-hydroxytryptamine and pineal metabolites on the secretion of neurohypophysial hormones. AB - It has been shown that 5-hydroxytryptamine and melatonin, an indoleamine for which 5-hydroxytryptamine is a precursor, influence the release of vasopressin and oxytocin from the rat hypothalamus both in vivo and in vitro. The oral administration of melatonin has been shown to decrease oxytocin release and modulate the nocturnal vasopressin release in humans. 5-hydroxytryptamine and its metabolites, 5-hydroxytryptophol, 5-methoxytryptamine and 5-methoxytryptophol, are detected within the pineal, and there is evidence that 5-methoxytryptamine and 5-methoxytryptophol may have some physiological role. The aim of this study was to evaluate the effects of 5-hydroxytryptamine, 5-hydroxytryptophol, 5 methoxytryptamine and 5-methoxytryptophol on neurohypophysial hormone release from the rat hypothalamus in vitro. It was found that 5-hydroxytryptamine and 5 hydroxytryptophol increased neurohypophysial hormone release, 5-methoxytryptamine decreased the release of vasopressin and oxytocin and 5-methoxytryptophol was found to have no effect, thus providing further evidence for a role of indole compounds in the control of neurohypophysial hormone secretion. PMID- 9739004 TI - Amyloid beta-peptides inhibit Na+/K+-ATPase: tissue slices versus primary cultures. AB - Abeta1-40 (20 microM) has been reported to selectively inhibit Na+/K+-ATPase activity in rat primary hippocampal cultures after 2-6 days of exposure. We expanded these studies to include Abeta's effects on Na+/K+-ATPase activity in rat primary cortical cultures and hippocampal slices, and we correlated these effects with estimates of cell survival in rat brain primary cultures. Using optimized assay conditions, a 5-day exposure to 50 microM Abeta 25-35, 20 microM Abeta 1-40, and 20 microM Abeta 1-42 decreased Na+/K+-ATPase activity in rat primary cortical cultures 66%, 60%, and 22%, respectively. Abeta 25-35 (50 microM) at 24 h was the only condition that caused inhibition of Na+/K+-ATPase activity in the absence of cell death, defined as an extracellular shift in the localization of the cytoplasmic enzyme lactate dehydrogenase (LDH). We also found that hippocampal slices were sensitive to Abeta, exhibiting a 40-60% reduction in membrane Na+/K+-ATPase activity when exposed to 1-30 nM of Abeta 1-40 for 60 min. This inhibition was not readily reversible, as it withstood homogenization and repeated dilution and centrifugation. Additionally, this inhibition occurred only after amyloid incubation with intact hippocampal slices, not with disrupted membranes. The inhibition of Na+/K+-ATPase in brain slices by physiological, low nM concentrations of Abeta 1-40 is consistent with effects on neurotransmitter release and intrasynaptosomal calcium responses. PMID- 9739005 TI - Molecular and immunochemical characterization of the ionotropic glutamate receptors in the rat heart. AB - Excitatory amino acids (EAA) and glutamate receptors (GluRs) play a fundamental role in the central nervous system (CNS). Ionotropic glutamate receptors (iGluRs) are coupled to ion channels, which are classified according to their most selective agonists. These ligand-gated channels are permeable to Na+, K+, and Ca+. Interaction of EAA receptor is linked to Ca+2/Na+ influx. Influx changes lead to an action potential, which in the heart is transmitted along the cardiocyte membrane. Furthermore, the heart has a rich innervation and specialized conduction system for rapid conduction and regulation of cardiac rhythmicity. Availability of EAA receptors in the heart might be important for cardiac function. The following GluRs were cloned by isoform-specific RT-PCR from rat heart ribonucleic acid (RNA): GluR 1, GluR 3, GluR 4, GIuR 7, Ka 1, and Ka 2. Expression in cardiac tissue was confirmed by western (for anti-GluR 2/3) and northern blots (for GluR 3, NMDAR 1, and Ka 2). The anatomical distribution was investigated by immunohistochemistry. Antibodies to GluR 2/3, GluR 5/6/7, Ka 2, and NMDAR 1 showed the strongest signals. These signals were specifically localized to cardiac nerve terminals, ganglia, conducting fibers, and some to myocardiocytes particularly in the atrium. Each antibody had a specific pattern of distribution. This anatomical localization suggests that they might play a role in cardiac electrophysiology and pathology. PMID- 9739006 TI - Cholinergic modulation of neuronal responses to cholecystokinin in anesthetized rats. AB - The aim of this study was to investigate whether the effects of the neuropeptide cholecystokinin on neuronal firing can be changed by acetylcholine in various structures of the brain. Single unit activity was extracellularly recorded in rats anesthetized with urethane. The neurons were located in several nuclei of the thalamus, the basal ganglia and the cerebral cortex. Neurons responding to the sulfated octapeptide of cholecystokinin (CCK-8S) were mainly activated by the drug [Wilcoxon test (Wt) p < 0.0001, n=113]. Thalamic neurons could also increase the number of burst discharges (Wt p < 0.005, n=39). Iontophoretically administered acetylcholine could reduce the activating effects of CCK-8S on firing and burst discharges. In its presence, even inhibitory effects of CCK-8S predominated (Wt p < 0.0001, n=113). The suppressive action seemed not to depend on the direction of the effect of acetylcholine itself and concerned neurons of all locations studied. Atropine could diminish or block the suppressive action of acetylcholine. In the presence of both drugs, CCK-8S mainly activated the neurons (Wt p < 0.005, n=43). Atropine itself did not significantly change the responses to CCK-8S (Wt p > 0.05). It can be concluded that cholecystokinin may reduce neuronal firing instead of increasing it during activation of the cholinergic system. PMID- 9739007 TI - Nicotine interacts with sex in affecting rat choice between "look-out" and "navigational" cognitive styles in the Morris water maze place learning task. AB - The effect of sex and nicotine on cognitive style was examined in rats using a water maze task that allows differentiation between cognitive ability and style. During the 12-day acquisition period with the platform in the same location (either visible or hidden) there were no effects or interactions attributable to nicotine and sex, either in terms of learning rate or asymptotic latency. On the final test day the platform was visible and shifted in its location, and on the first trial the new location was proximal to the rats starting position, in contrast to the more distal location of the platform during the previous acquisition days. This platform relocation presented the rats with a choice between two competing cognitive styles: using local visual (look-out) cues vs. navigational cues. Performance on the test day yielded a nicotine x sex interaction, such that only saline-treated female rats showed a clear preference for the perceptual-proximal look-out cognitive style by swimming straight to the newly-relocated visible platform with mean escape latency that approximated the limits of swimming speed. The other three groups did not differ from each other, and preferred navigational cues. The results show that male and female rats use different strategies in problem solving, and that nicotine shifts the female pattern to that of the male. PMID- 9739008 TI - Different effects of insulin and 2-deoxy-D-glucose administration on tyrosine hydroxylase gene expression in the locus coeruleus and the adrenal medulla in rats. AB - The major brain norepinephrinergic nucleus, locus coeruleus, is an important integrating element of extero- and interoceptive stimuli in organisms facing different physiological challenges. We investigated the effects of single and repeated (seven times) exposure to immobilization stress (120 min daily), insulin (5 IU/kg, i.p. daily) or 2-deoxy-D-glucose (500 mg/kg, i.p. daily) administration on tyrosine hydroxylase (TH) mRNA levels, the rate-limiting enzyme in catecholamine biosynthesis, by in situ hybridization in locus coeruleus and by Northern blot analysis in the adrenal medulla of rats. Both the single and repeated immobilization caused a significant increase in TH mRNA levels in the locus coeruleus (1.5-2-fold; p < 0.05) and in the adrenal medulla (about 4-fold; p < 0.05) when compared with unstressed controls. Hypoglycemia induced by a single or repeated insulin administration led to about fourfold (p < 0.01) elevation in adrenal medullary TH mRNA levels, whereas TH mRNA in locus coeruleus remained unchanged when compared with saline-treated controls. In contrast to the effect of insulin-induced hypoglycemia, cellular glucoprivation caused by a single or repeated 2-deoxy-D-glucose administration significantly elevated TH mRNA levels in both the adrenal medulla (fourfold; p < 0.01) and the locus coeruleus (twofold; p < 0.01). Our data suggest that in contrast to immobilization or cellular glucoprivation caused by 2-deoxy-D-glucose administration, insulin-induced hypoglycemia is not a specific or quantitatively sufficient stimulus for induction of TH gene expression in the locus coeruleus, although all these stressors strongly activate the process in the adrenal medulla. PMID- 9739009 TI - Dual projection from the medial septum to the supramammillary nucleus in the rat. AB - The supramammillary nucleus, collecting information about the physiological state of the animal, innervates medial septal neurons that are involved in the generation of hippocampal theta activity. Here we demonstrate that septal neurons located in an area bordering the medial and lateral septal nucleus project back to the supramammillary nucleus, and most of these cells contain calretinin, calbindin or both. GABA-immunoreactive boutons of these neurons (60%) form symmetrical synapses, whereas the remaining GABA-negative terminals form asymmetrical synapses (40%) with their supramammillary targets. We hypothesize that the septosupramammillary feedback, because of the specific location of its parent cells, carries information about the activity of theta generator cells in the medial septum and supramammillary nucleus, as well as about the resulting theta activity in the hippocampus. PMID- 9739010 TI - The effects of histamine H3-receptor antagonists on amygdaloid kindled seizures in rats. AB - The effects of histamine H3-receptor antagonists, thioperamide, and clobenpropit on amygdaloid kindled seizures were investigated in rats. Both intracerebroventricular (i.c.v.) and intraperitoneal (i.p.) injections of H3 antagonists resulted in a dose-related inhibition of amygdaloid kindled seizures. An inhibition induced by thioperamide was antagonized by an H3-agonist [(R)-alpha methylhistamine] and H1-antagonists (diphenhydramine and chlorpheniramine). On the other hand, an H2-antagonist (cimetidine and ranitidine) caused no antagonistic effect. Metoprine, an inhibitor of N-methyltransferase was also effective in inhibiting amygdaloid kindled seizure, and this effect was augmented by thioperamide treatment. PMID- 9739011 TI - Cytogenetic analysis of pancreatic carcinomas: intratumor heterogeneity and nonrandom pattern of chromosome aberrations. AB - Twenty-nine nonendocrine pancreatic carcinomas (20 primary tumors and nine metastases) were studied by chromosome banding after short-term culture. Acquired clonal aberrations were found in 25 tumors and a detailed analysis of these revealed extensive cytogenetic intratumor heterogeneity. Apart from six carcinomas with one clone only, 19 tumors displayed from two to 58 clones, bringing the total number of clones to 230. Karyotypically related clones, signifying evolutionary variation, were found in 16 tumors, whereas unrelated clones were present in nine, the latter finding probably reflecting a distinct pathogenetic mechanism. The cytogenetic profile of pancreatic carcinoma was characterized by multiple numerical and structural changes. In total, more than 500 abnormal chromosomes, including rings, markers, homogeneously stained regions, and double minutes, altogether displaying 608 breakpoints, were detected. This complexity and heterogeneity notwithstanding, a nonrandom karyotypic pattern can be discerned in pancreatic cancer. Chromosomes 1, 3, 6, 7, 8, 11, 12, 17, and 19 and bands 1q12, 1q21, 3q11, 6p21, 6q21, 7q11, 7q22, 7q32, 11q13, 13cen, 14cen, 17q11, 17q21, and 19q13 were most frequently involved in structural rearrangements. A total of 19 recurrent unbalanced structural changes were identified, 11 of which were not reported previously: del(1)(q11), del(3)(p11), i(3)(q10), del(4)(q25), del(11)(p13), dup(11)(q13q23), i(12)(p10), der(13;15)(q10;q10), del(18)(q12), del(18)(q21), and i(19)(q10). The main karyotypic imbalances were entire-copy losses of chromosomes 18, Y, and 21, gains of chromosomes 7, 2, and 20, partial or whole-arm losses of 1p, 3p, 6q, 8p, 9p, 15q, 17p, 18q, 19p, and 20p, and partial or whole-arm gains of 1q, 3q, 5p, 6p, 7q, 8q, 11q, 12p, 17q, 19q, and 20q. In general, the karyotypic pattern of pancreatic carcinoma fits the multistep carcinogenesis concept. The observed cytogenetic heterogeneity appears to reflect a multitude of interchangeable but oncogenetically equivalent events, and the nonrandomness of the chromosomal alterations underscores the preferential pathways involved in tumor initiation and progression. PMID- 9739012 TI - Characterization of recurrent homogeneously staining regions in 72 breast carcinomas. AB - Cytogenetic analyses were performed on 223 breast carcinomas, of which 60% contained homogeneously staining regions (hsr), an intrachromosomal cytogenetic feature of gene amplification. The precise hsr localization could be determined for 123 hsr from 72 cases. The juxtacentromeric region of chromosome 8, band 11q13, and the whole of chromosome 17 were frequently involved. For 28 cases, the origin of the DNA sequences forming HSR could be investigated by chromosome painting, comparative genomic hybridization, and/or Southern blotting. Sequences from chromosomes 11 and 17 were mostly found within hsr located on chromosomes 11 and 17, respectively. In contrast, sequences from chromosome 8 were rarely found within hsr localized on chromosome 8. These observations suggest that different mechanisms lead to hsr formation in breast cancer. Band 11 q13 and the 17p chromosome arm may correspond to sites of in situ amplification driven by deletions distal to the amplification target genes. hsr in the region 17q2, which is also a frequent site of in situ amplification, takes place without the occurrence of a distal deletion. The short arm of chromosome 8 is often deleted, but frequently becomes the site of hsr formed elsewhere in the genome. PMID- 9739013 TI - A two-color BCR-ABL probe that greatly reduces the false positive and false negative rates for fluorescence in situ hybridization in chronic myeloid leukemia. AB - The t(9;22) translocation resulting in the fusion of BCR and ABL genes is pathognomonic in chronic myeloid leukemia (CML) and may be investigated at the molecular level using fluorescence in situ hybridization (FISH). Two-color BCR ABL probes visualizing one fusion signal (1F FISH) have high false positive rates (FPR) and false negative rates (FNR). The FPR is a result of the random spatial association of probe signals within normal interphase cells so that some cells appear to contain the BCR-ABL fusion gene. The FNR of 1F FISH probes depends on the distance between the BCR and ABL probes hybridized to the BCR-ABL fusion gene (< or =368 kb); the "gap" between the signals causing the cell to be interpreted as normal. To overcome these difficulties, a two-color probe was used, employing four yeast artificial chromosome (YAC) sequences that span the breakpoint regions of the BCR and ABL genes and that visualize the two fusion signals BCR-ABL and ABL-BCR in CML cells (2F FISH). The FNR for the 2F FISH probes was assessed on clonal Ph+ granulocyte-macrophage-colony-forming cell (CFU-GM) derived colonies and was reduced to 0.4% (2/450), compared with an FNR of 13.5% (111/823) with 1F FISH. The FPR in normal mononuclear cells for the 2F FISH was 0. 19 +/- 0.12% (3/1,700), whereas the FPR using 1F FISH was 4.5 +/- 2.3% (63/1,294). The 2F FISH can thus be used to evaluate very small frequencies of BCR-ABL-positive and negative interphase cells and may be of use in the clinical monitoring of CML. PMID- 9739014 TI - Molecular cytogenetic delineation of 17q translocation breakpoints in neuroblastoma cell lines. AB - It has recently been recognized that unbalanced translocations resulting in the gain of material from 17q are the most common chromosomal changes in neuroblastoma. These rearrangements are associated with established indicators of bad prognosis and poor patient survival. We have used 13 fluorescence in situ hybridization (FISH) probes to map 12 translocation breakpoints on 17q in 10 neuroblastoma cell lines, identifying at least seven different breakpoints, all localized within the proximal half of 17q (268-369 cR, 53-68 cM). These results suggest that the dosage of a gene, or genes, in 17q22-qter is responsible for the clinical effects of 17q gain, rather than the disruption of a specific gene. This region contains two genes, nm23-H1 and NGFR, already implicated in neuroblastoma biology. PMID- 9739015 TI - Evidence for a putative telomerase repressor gene in the 3p14.2-p21.1 region. AB - Telomeres, which are the repeated sequences located on both ends of chromosomes in eukaryotes, are known to shorten with each cell division, and their eventual loss is thought to result in cellular senescence. Unlike normal somatic cells, most tumor cells show activation of telomerase, a ribonucleoprotein enzyme that stably maintains telomere length by addition of the sequences of TTAGGG repeats to telomeres. The KC12 cell line derived from a renal cell carcinoma in a patient with von Hippel-Lindau disease showed telomerase activity and loss of heterozygosity on the short arm of chromosome 3. Introduction of a normal human chromosome 3 into KC12 cells by microcell fusion induced cellular senescence, accompanied by suppression of telomerase activity and shortening of telomere length. Microcell hybrids that escaped from cellular senescence maintained telomere length and telomerase activity similar to those of the parental KC12 cells. We previously showed a similar suppression of telomerase activity by introduction of chromosome 3 into another renal cell carcinoma cell line, RCC23. The putative telomerase repressor gene was mapped to chromosome region 3p14.2 p21.1 by deletion mapping of KC12 + chromosome 3 revertants that escaped from cellular senescence and by transfer of subchromosomal fragments of chromosome 3 into RCC23 cells. PMID- 9739016 TI - MYCN is the only highly expressed gene from the core amplified domain in human neuroblastomas. AB - MYCN amplification in neuroblastomas is strongly associated with advanced stages of disease and a poor prognosis. We have recently defined a 130 kb core region of the MYCN amplicon that is consistently amplified in neuroblastomas. However, it has been argued that other expressed sequences were coamplified with MYCN and, as a result, might contribute to the aggressive phenotype of MYCN-amplified neuroblastomas. Therefore, we have screened cosmids representing the core MYCN amplified domain and surrounding DNA by using a differential hybridization approach to detect other amplified, highly expressed genes from this region. Our results suggest that MYCN is the only highly expressed gene consistently amplified in human neuroblastomas, and that the MYCN gene is likely to be the only selective marker for genomic amplification in these tumors. PMID- 9739018 TI - Dynamics of genetic alterations associated with glioma recurrence. AB - We investigated the dynamics of the genetic changes that are associated with two types of glioma recurrence, that is, progression from a lower-grade to a high grade tumor (7 cases) and development of a same high-grade recurrence (15 cases). Each pair of tumors was analyzed for TP53 mutation, EGFR amplification, and loss of heterozygosity for tumor suppressor genes (TP53, RB1, CDKN2A, PTEN, DMBT1) and tumor suppressor gene regions (1p36, 19q13, 11p15, 10p15) known to be frequently implicated in glioma tumorigenesis. By comparing the genetic changes in the primary and corresponding secondary tumors, we found that additional loss of CDKN2A and/or RB1, encoding important components of the cell cycle regulatory pathway, was the most frequent genetic change in both types of recurrence development (10 of 22 cases, 45%). Additional loss of heterozygosity for the 10p15 region, for PTEN, and/or for DMBT1 in the recurrent tumor was noted in 7 of 22 cases (32%), suggesting that additional inactivation of tumor suppressor genes on chromosome 10 is another important feature of glioma relapse. Less frequent additional losses were detected for chromosome regions 11p15 and 19q13 (3 of 22 cases, 14%, each). We conclude that glioma recurrences are characterized by an increased involvement of tumor suppressor genes, even in those cases in which the primary and secondary tumor are of the same high malignancy grade. PMID- 9739017 TI - Genetic heterogeneity of neuroblastoma studied by comparative genomic hybridization. AB - Comparative genomic hybridization (CGH) analysis was performed on 36 neuroblastomas of both low and high stage of disease. This study significantly increases the number of neuroblastoma tumors studied by CGH. Analysis of larger series of tumors is particularly important in view of the different clinical subgroups that are recognized for this tumor. The present data and a comparison with all published CGH data on neuroblastoma provide further insights into the genetic heterogeneity of neuroblastoma. Stage 1, 2, and 4S tumors showed predominantly whole chromosome gains and losses. A similar pattern of whole chromosome imbalances, although less frequent, was observed in stage 3 and 4 tumors, in addition to partial chromosome gains and losses. An increase in chromosome 17 or 17q copy number was observed in 81% of tumors. The most frequent losses, either through partial or whole chromosome underrepresentation, were observed for 1p (25%), 3p (25%), 4p (14%), 9p (19%), 11q (28%), and 14q (31%). The presence of 3p, 11q or 14q deletions defines a genetic subset of neuroblastomas and contributes to the further genetic characterization of stage 3 and 4 tumors without MYCN amplification (MNA) and 1p deletion. The present study also provides additional evidence for a possible role of genes at 11q13 in neuroblastoma. In a few cases, 1p deletion or MNA detected by FISH or Southern blotting was not found by CGH, indicating that the use of a second, independent technique for evaluation of these genetic parameters is recommended. PMID- 9739019 TI - Mutation of the mismatch repair gene hMSH2 and hMSH6 in a human T-cell leukemia line tolerant to methylating agents. AB - Cell killing by monofunctional methylating agents is due mainly to the formation of adducts at the O6 position of guanine. These methyl adducts are removed from DNA by the O6-alkylguanine DNA alkyltransferase (OGAT). The mechanism by which O6 methylguanine (O6meG) induces cell death in OGAT-deficient cells requires a functional mismatch repair system (MRS). We have previously reported that depletion of OGAT activity in the human T-cell leukemic urkat line does not sensitize these cells to the cytotoxic and apoptotic effects of the methylating triazene temozolomide (Tentori et al., 1995). We therefore decided to establish whether the tolerance of Jurkat cells to O6meG could be associated with a defect in MRS. The results of mismatch repair complementation studies indicated that Jurkat cells are defective in hMutSalpha, a heterodimer of the hMSH2 and hMSH6 proteins. Cytogenetic analysis of two Jurkat clones revealed a deletion in the short arm of chromosome region 2p15-21, indicating an allelic loss of both hMSH2 and hMSH6 genes. DNA sequencing revealed that exon 13 of the second hMSH2 allele contains a base substitution at codon 711, which changes an arginine to a termination codon (CGA-->TGA). In addition, a (C)8-->(C)7 frameshift mutation in codon 1085-1087 of the hMSH6 gene was also found. Although both hMSH2 and hMSH6 transcripts could be detected in Jurkat clones, the respective polypeptides were absent. Taken together, these data indicate that tolerance of Jurkat cells to methylation damage is linked to a loss of functional hMutSalpha. PMID- 9739020 TI - Chromosomal changes during progression of transitional cell carcinoma of the bladder and delineation of the amplified interval on chromosome arm 8q. AB - The cascade of genetic alterations leading to malignant transformation has been described for adenocarcinoma of the colon but is not established for other common tumor entities. In the present study, different stages of transitional cell carcinoma (TCC) of the bladder are analyzed by comparative genomic hybridization. A dynamic pattern of the chromosomal changes during tumor progression is described. Deletion of chromosome arm 9q is the earliest genetic alteration in pTa tumors. In stage pT1 carcinomas, losses of 9q, 9p, and 11p and gain of 1q and 8q are the most common. In addition to the changes specific for earlier stages, gain of 5p and 20q becomes prominent in carcinomas stage > or =pT2. Association analysis reveals a remarkable cooccurrence of 9p deletion with gain of 5p and 20q in > or =pT2 tumors. In order to determine more precisely the size of the amplified segment and the degree of amplification on chromosome arm 8q in stage pT1 tumors, this region was analyzed by semiquantitative PCR using polymorphic microsatellite markers. These studies revealed an up to 13-fold amplification. The common region of amplification could be narrowed down to 8q22.3 and between GAAT1A4 and D8S1834 (about 7 cM). PMID- 9739021 TI - Detection of t(11;14) using interphase molecular cytogenetics in mantle cell lymphoma and atypical chronic lymphocytic leukemia. AB - The chromosomal translocation t(11;14)(q13;q32) fuses the IGH and CCND1 genes and leads to cyclin D1 overexpression. This genetic abnormality is the hallmark of mantle cell lymphoma (MCL), but is also found in some cases of atypical chronic lymphocytic leukemia (CLL), characterized by a poor outcome. For an unequivocal assessment of this specific chromosomal rearrangement on interphase cells, we developed a set of probes for fluorescence in situ hybridization (FISH). Northern blotting was performed for analysis of the cyclin D1 expression in 18 patients. Thirty-eight patients, with either a typical MCL leukemic phase (17 patients) or atypical CLL with an MCL-type immunophenotype, i.e., CD19-, CD5+, CD23-/low, CD79b/sIgM(D)++, and FMC7+ (21 patients), were analyzed by dual-color interphase FISH. We selected an IGH-specific BAC probe (covering the JH and first constant regions) and a commercially available CCND1 probe. An IGH-CCND1 fusion was detected in 28 of the 38 patients (17 typical MCL and 11 cases with CLL). Cyclin D1 was not overexpressed in two patients with typical MCL and an IGH-CCND1 fusion. In view of the poor prognosis associated with MCL and t(11;14)-positive CLL, we conclude that this set of probes is a valuable and reliable tool for a rapid diagnosis of these entities. PMID- 9739022 TI - Chromosome band 1q21 is recurrently gained in desmoid tumors. AB - DNA sequence copy number changes were studied by comparative genomic hybridization (CGH) in 28 desmoid tumors. Changes were detected in 12 tumors (43%) with a mean of 1.4 changes per sample (range: 1 to 7). Out of 12 tumors associated with pregnancy or Gardner's syndrome, only two displayed changes. The minimal common regions of the most frequent gains were 1q21 (39%), chromosome 20 (32%), and 9p12 (21%). No high-level amplifications were detected. Losses of DNA sequences were two times less frequent than gains and the minimal common regions of the most frequent losses were 6q16-q21 (14%), 5q14 (11%), and 13q21-q31 (11%). PMID- 9739024 TI - Alveolar soft-part sarcoma: further evidence by FISH for the involvement of chromosome band 17q25. AB - A cytogenetic study of an alveolar soft-part sarcoma, a rare tumor of probably myogenic origin, demonstrated a t(X;17)(p11;q25) as the sole chromosomal abnormality. Dual- and triple-color fluorescence in situ hybridization, performed on metaphase and interphase cells, confirmed the translocation between chromosomes X and 17 and demonstrated that this translocation resulted in loss of 17q25. Involvement of 17q25 has been described in four previously published cases of alveolar soft-part sarcoma, but without further characterization. Compared to our karyotype, it seems that the derivative chromosome 17 observed in the reported cases could also be the result of a t(X;17) with possible loss of the 17q25 band. If so, a 17q25 deletion and/or chromosome rearrangement between Xp and 17q leading either to a gene fusion or gene disruption could play an important role in the pathogenesis of alveolar soft-part sarcoma. PMID- 9739023 TI - Various regions within the alpha-helical domain of the COL1A1 gene are fused to the second exon of the PDGFB gene in dermatofibrosarcomas and giant-cell fibroblastomas. AB - Dermatofibrosarcoma protuberans (DFSP) and its juvenile form, giant-cell fibroblastoma (GCF), are uncommon infiltrative tumors of the dermis, which present unique cytogenetic features, such as the reciprocal translocation t(17;22) or, more commonly, supernumerary ring chromosomes containing sequences from chromosomes 17 and 22. We have recently shown that these aberrations are cytogenetic manifestations of gene fusions between the platelet-derived growth factor B-chain gene (PDGFB), the cellular equivalent of the v-sis oncogene, and the collagen type 1 alpha 1 gene (COL1A1), the major protein constituent of the extracellular matrix in connective tissue of skin. We now report characterization of COL1A1/PDGFB chimeric genes at the RNA and DNA sequence levels in a series of DFSPs and GCFs. All 16 tumors studied contained the COL1A1/PDGFB gene. The location of breakpoints within COL1A1 varied greatly, but was always limited to the region encoding the alpha-helical domain. The PDGFB segment of the chimeric transcript always starts with exon 2, placing PDGFB under the control of the COL1A1 promoter and removing all known elements negatively controlling PDGFB transcription and translation. Production of these aberrant transcripts in fibroblasts, the suspected cell of origin of DFSP/GCF, likely causes autocrine stimulation and cell proliferation. No specific function has yet been assigned to exon 2 of PDGFB, and this exon does not encode for the mature growth factor. Its retention in all chimeric COL1A1/PDGFB genes suggests that it is important for the normal processing of the PDGFB polypeptide. PMID- 9739025 TI - Masked t(X;18)(p11;q11) in a biphasic synovial sarcoma revealed by FISH and RT PCR. AB - The initial cytogenetic analysis of a biphasic synovial sarcoma showed an apparently normal karyotype. After FISH using chromosome X- and 18-specific probes and RT-PCR using SYT- and SSX-specific primer sets, a cryptic synovial sarcoma-associated t(X;18)(p11;q11) could be revealed. The "masked" nature of the translocation may best be explained by a two-step scenario in which a genuine t(X;18)(p11;q11) has occurred as a first step and a reverse reciprocal X;18 translocation as a second step, leaving the synovial sarcoma-associated SYT-SSX1 fusion intact. The findings further underline our previous suggestion that SYT SSX1 fusions may correlate with a biphasic nature of the tumor. In addition, our findings indicate that, in analogy to, e.g., the Philadelphia translocation in chronic myeloid leukemia, "masked" translocations may occur in soft tissue tumors and that, as a standard, RT-PCR and/or FISH analyses should be carried out in order to provide karyotypic information that may be relevant to tumor diagnosis and/or prognosis. PMID- 9739026 TI - The influence of intravenous infusions of glucose and amino acids of pancreatic enzyme and mucosal protein synthesis in human subjects. AB - BACKGROUND: Animal studies have shown that the synthesis and secretion of pancreatic enzymes and the turnover of mucosal proteins is strongly influenced by diet. METHODS: To determine whether the absorbed products of digestion are responsible for these changes, we investigated in groups of five healthy volunteers, the effects of i.v. infusions of amino acids (0.08 g/kg/h) and glucose (0.3 g/kg/h) on pancreatic enzyme and mucosal protein synthesis. Proteins were labeled in vivo by a 4-hours i.v. infusion of 14C-leucine and the enteric infusion of 3H-leucine tracer, during simultaneous cholecystokinin stimulation and duodenal collection of secreted pancreatic enzymes. Labeling of mucosal proteins was measured by endoscopic biopsy. RESULTS: The amino acid infusions elevated plasma amino acid levels, and the glucose infusions increased both glucose and insulin concentrations. The rates for amylase and trypsin secretion were significantly lower during the first 2 hours of glucose infusion and the rate of synthesis of trypsin was delayed by i.v. amino acid infusions from 52.1 +/- 4.1 to 77.6 +/- 8.5 minutes. Mucosal protein turnover rates were unaffected. 3H-labeling via the enteral route showed similar enzyme synthesis rates but variable mucosal incorporation rates. CONCLUSIONS: I.v. nutrients do not appear to stimulate the synthesis of pancreatic and mucosal proteins in human subjects. PMID- 9739027 TI - Gut-trophic effects of keratinocyte growth factor in rat small intestine and colon during enteral refeeding. AB - BACKGROUND: Keratinocyte growth factor (KGF) induces proliferation of gut epithelium in rat models, but KGF-nutrient interactions have not been studied. An experimental model of fasting-induced gut atrophy followed by different levels of enteral refeeding was used to investigate the influence of nutrient availability on the gut-trophic effects of exogenous KGF. METHODS: After a 3-day fast, rats were enterally refed either ad libitum or at 25% of ad libitum intake for 3 subsequent days. Either intraperitoneal KGF (5 mg/kg/d) or saline was given in each dietary regimen. Wet weight, DNA, and protein content were measured as indices of full-thickness cellularity in duodenum, jejunum, ileum, and colon. Villus height in small bowel segments and crypt depth in all gut tissues were measured as specific indices of mucosal growth. RESULTS: Refeeding at 25% of ad libitum intake significantly decreased full-thickness cellularity and mucosal growth indices in duodenum, jejunum, and ileum. In the colon, only protein content fell significantly and crypt depth was maintained. KGF administration during 25% refeeding did not alter full-thickness indices in any small bowel segment or affect jejunal mucosal growth. In contrast, KGF normalized duodenal villus height (p < .01) and duodenal and ileal crypt depth (p < .05) only in the 25%-refed model. KGF significantly increased ileal villus height in both ad libitum and 25%-refed rats (by 43% and 48%, respectively, p < .05) and markedly increased colonic cellularity and mucosal crypt depth with both levels of refeeding (p < .01). CONCLUSIONS: Rat small bowel growth is more sensitive than colon to the level of enteral refeeding after a 3-day fast. KGF administration does not affect jejunal growth, but specifically prevents atrophy of duodenal and ileal mucosa during hypocaloric, hyponitrogenous refeeding. In ileum and colon, some KGF-mediated growth responses are independent of the level of enteral refeeding. Thus gut-trophic effects of KGF and KGF interactions with the level of nutrient intake are tissue-specific. PMID- 9739028 TI - Role of arachidonic acid in the regulation of the inflammatory response in TNF alpha-treated rats. AB - BACKGROUND: This study examined whether adding arachidonic acid (AA) to a fish oil diet would alter certain of the anti-inflammatory effects of fish oil in response to tumor necrosis factor (TNF) infusion in rats. METHODS: AA was given at 0.08 wt% of diet for 6 weeks. The total fat in each diet provided 20% of dietary energy. Four groups were pair-fed sunflower oil (S), S+AA, fish oil (F), or F+AA for 6 weeks. At the end of feeding, each animal received TNF-alpha (20 microg/kg) infusion for 3 hours. After 1 hour of TNF infusion, a euglycemic and hyperinsulinemic clamp (10 mU/min per kilogram of insulin) was used to determine the actions of insulin. The insulin-stimulated glucose utilization in liver, muscle, and fat was determined by using 14C-deoxyglucose. The plasma glucose, insulin, and corticosterone levels were determined at basal, 60 minutes, and the end of the experiment (180 minutes). The fatty acid composition of plasma phospholipids also was determined. RESULTS: Fish oil significantly increased omega-3 fatty acids in phospholipids in both F and F+AA and decreased AA in F, compared with S. AA significantly restored the level of AA and reduced the increase of omega-3 fatty acids in phospholipids in F+AA compared with F, but had no impact on fatty acid composition when added to S. Corticosterone level was significantly lower with fish oil feeding but higher in both F and S containing AA compared with F and S, respectively. The highest glucose uptake in tissues was in F, followed by F+AA, and then S and S+AA. CONCLUSIONS: These results suggest that fish oil is anti-inflammatory principally through a reduction in the AA content of phospholipids. PMID- 9739029 TI - The route of nutrition support affects the early phase of wound healing. AB - BACKGROUND: Nutrition support via the enteral route has been shown to be superior to parenteral administration in maintaining immune function, decreasing septic complications, and increasing survival after severe trauma and surgical injury. Whether the route of nutrition support affects wound healing, another important determinant of outcome following injury, is not known. METHODS: Forty-nine Sprague-Dawley rats, 290 to 360 g body wt, underwent identical surgical manipulation consisting of central venous catheterization, fashioning of gastrostomy and dorsal skin incision, and placement of polyvinyl alcohol sponges into subcutaneous pockets. Identical infusates of 25% dextrose, 4.25% amino acids, and vitamins were given, half the animals receiving the infusion via the gastrostomy and the other half via the venous catheter. Animals were killed on day 5, 7, or 10. Wound breaking strength, sponge hydroxyproline content (an index of wound collagen deposition), and types I and III collagen gene expression were measured. RESULTS: There were no nutritional differences between the two groups in terms of energy intake, body weight gain, and plasma levels of albumin, total protein, or urea nitrogen. On day 5 wound breaking strength was significantly higher in the enterally supported group (89.3 +/- 90.7 vs 64.9 +/- 40.2 g for the parenteral group, p < .05). This was paralleled by enhanced wound collagen accumulation (182 +/- 19 vs 132 +/- 13 microg, p < .05). Gene expression of type I, but not type III, collagen also was increased in the enterally fed group. There were no differences noted between the two groups in wound healing parameters 7 and 10 days after injury. CONCLUSIONS: The data demonstrate that the route of nutrition administration can influence wound healing. The beneficial effect of the enteral feeding route is limited to the early phases of healing. PMID- 9739030 TI - Parenteral glutamine infusion alters insulin-mediated glucose metabolism. AB - BACKGROUND: Glutamine is a conditionally essential amino acid that is critical for many basic cellular processes. Its supplementation has been found to be beneficial during several critical illnesses. This study examines the effects of increased glutamine availability on insulin-mediated glucose homeostasis in vivo in multicatheterized conscious canines (n = 5). METHODS: Two weeks before the study, catheters were placed in the femoral artery and the portal, hepatic, femoral, and renal veins for blood sampling and in the splenic vein for intraportal infusion of insulin and glucagon. Doppler probes were placed to measure blood flow. The metabolic study consisted of equilibration, basal, and experimental periods during which [3-3H]glucose was infused to measure glucose kinetics. During the 5-hour experimental period, a hyperinsulinemic-euglycemic clamp was performed by infusing somatostatin, basal glucagon, fivefold basal insulin, and glucose to maintain euglycemia. The experimental period was divided evenly into two subperiods performed in random order: (1) i.v. glutamine infusion (0.72 mmol kg(-1) h(-1)) and (2) i.v. saline infusion. RESULTS: With glutamine, the glucose required to maintain euglycemia was increased 46% over saline (6.8 +/ 1.0 to 9.9 +/- 1.7 mg kg(-1) min(-1). In addition, whole-body glucose production and utilization were increased by 1.4 and 4.6 mg kg(-1) min(-1), respectively. Finally, the increase in whole-body glucose utilization was manifested by increased hepatic and hindlimb glucose utilization. CONCLUSIONS: Increased glutamine availability blunted insulin's action on glucose production and enhanced insulin-mediated glucose utilization with the changes in utilization being threefold greater than the changes in production. Thus parenteral glutamine has potential benefit as a nutrient adjuvant during clinical situations associated with insulin resistance. PMID- 9739031 TI - TNF-alpha and IL-6 synergistically inhibit ketogenesis from fatty acids and alpha ketoisocaproate in isolated rat hepatocytes. AB - BACKGROUND: During sepsis, lipid metabolism is shunted toward triacylglycerol synthesis and hepatic lipogenesis. A decrease in ketogenesis from free fatty acids also is observed, probably mediated by cytokines involved in host response to infection. Whether such an inhibition of ketogenesis occurs with other ketone body precursors such as ketoacids is not known. The aim of this study was to determine the effects of tumor necrosis factor alpha (TNF-alpha) and interleukin 6 (IL-6) on hepatic ketone body production from octanoic acid, a medium-chain fatty acid, and from alpha-ketoisocaproate (KIC), the ketoanalogue of leucine. METHODS: The experiments were conducted in cultured hepatocytes isolated from 24 hour-fasted Sprague-Dawley rats. Hepatocyte monolayers were incubated for 6 hours, with either KIC or octanoic acid (1 mmol/L), in the presence of glucagon and TNF-alpha (25 micro/L) IL-6 (15 microg/L) and/or IL-6. Acetoacetate, beta hydroxybutyrate, and free fatty acids were determined in culture medium by enzymatic methods and KIC was measured by high-performance liquid chromatography. RESULTS: KIC and octanoic acid uptake by hepatocytes was 79% and 92%, respectively, over 6 hours, and cytokines had no influence. However, TNF-alpha and IL-6 caused inhibition of ketogenesis from alpha-ketoisocaproate (5.6% +/- 2.3% and 4.4% +/- 3.0%, respectively), and from octanoic acid (7.9% +/- 2.9%, 5.7% +/- 3.2%, respectively). In addition, when the two cytokines were present together in the culture medium, the inhibition was enhanced (inhibition of ketogenesis from KIC: 14.0% +/- 4.8%; from octanoic acid: 11.6% +/- 3.4%). CONCLUSIONS: In our experimental conditions, cytokines mediate an inhibition of ketogenesis; this process could be explained by a direct effect of cytokines on metabolic pathways of octanoic acid and KIC oran indirect effect by modification of the mitochondrial redox state. PMID- 9739032 TI - Total nutrient admixtures appear safer than lipid emulsion alone as regards microbial contamination: growth properties of microbial pathogens at room temperature. AB - BACKGROUND: The extraordinary growth properties of most microorganisms in 10% and 20% lipid emulsions has led to the Centers for Disease Control and Prevention recommendation that if lipids are given through an i.v. line, the administration set should be replaced every 24 hours rather than the usual 72-hour interval used for crystalloid solutions, including those used for conventional total parenteral nutrition. For nearly 15 years, parenteral alimentation has been given as a total nutrient admixture (TNA), with the glucose, amino acids, and lipid mixed within the same bag and infused continuously over 24 hours. METHODS: We prospectively studied in a representative TNA (17.6% glucose, 5% amino acids, 4% lipid; pH 5.6, osmolality 1778) and in a control solution, 5% dextrose-in-water (D5%/W), the growth properties at 4, 25, and 35 degrees C of three isolates each of Staphylococcus epidermidis, Staphylococcus aureus, Enterobacter cloacae, Klebsiella oxytoca, Serratia marcescens, Acinetobacter calcoaceticus, Stenotrophomonas maltophilia, Pseudomonas aeruginosa, Burkholderia cepacia, Flavobacterium spp, and Candida albicans, and two isolates of Staphylococcus saprophyticus, the species that are most likely to contaminate TNA during preparation or administration and that have been implicated in >95% of all outbreaks and sporadic cases of nosocomial bloodstream infections traced to contaminated parenteral admixtures reported in the world literature. RESULTS: Growth in TNA at 25 and 35 degrees C occurred with only two species, C. albicans and S. saprophyticus, and only after 24 to 48 hours; D5%/W allowed growth at 25 degrees C of two gram-negative species, S. marcescens and B. cepacia. CONCLUSIONS: We conclude that TNA is a poor growth medium for most nosocomial pathogens and is no better than D5%/W. The need to replace administration sets every 24 hours with TNA should be reconsidered and ideally be studied in a prospective randomized trial. PMID- 9739033 TI - Effects of soybean oil and fish oil emulsions on glucose and lipid metabolism in streptozotocin-induced diabetic rats receiving total parenteral nutrition. AB - BACKGROUND: This study was designed to investigate the effects of fat emulsions with different fatty acid composition on plasma glucose and lipid metabolism in diabetic rats receiving total parenteral nutrition (TPN). METHODS: Diabetes was induced in rats with streptozotocin (STZ), and the rats were fed rat chow ad libitum for 6 weeks to achieve a chronic diabetic state. Control and diabetic rats were each divided into two TPN groups. The basal solutions of the two TPN groups were isonitrogenous and identical in nutrients composition except for the fat emulsion, which was made of soybean oil (SO) or fish oil (FO). The TPN control rats (C-SO and C-FO) and diabetic rats (DM-SO and DM-FO) received solutions with 37.5% of the non-protein energy provided as fat at an energy level of 30 kcal/100 g body wt/d. RESULTS: The results demonstrated that hyperglycemia and hypertriglyceridemia were induced by STZ in diabetic rats. There was no change in plasma glucose and insulin concentrations before and after TPN infusion in the TPN control groups, whereas plasma glucose as well as triglyceride (TG) and nonesterified fatty acid (NEFA) levels decreased significantly after TPN administration in the diabetic groups. No difference in the concentrations of plasma glucose, TGs, NEFAs, and insulin were observed between the two diabetic groups. CONCLUSIONS: These results suggest that compared with soybean oil, TPN with fish oil emulsion did not lead to lower plasma concentrations of TGs and NEFAs in STZ-induced diabetic rats. Also, no difference in plasma glucose and insulin levels between the two groups was observed. PMID- 9739034 TI - Cost and outcome analysis of home parenteral and enteral nutrition. AB - BACKGROUND: Previous estimates of the cost of home parenteral and enteral nutrition (HPEN) have excluded hospitalization costs or were conducted abroad and have limited applicability in the United States. Few studies have used validated measures to determine the effect of home nutrition support on quality of life. METHOD: A cost and clinical outcome analysis was performed by retrospective review of charts of patients receiving HPEN from 1991 to 1996. Questionnaires to determine the influence of therapy on lifestyle (n = 41) and a general health status questionnaire, the short form 36-item survey (n = 39), were mailed to patients. RESULTS: The annual cost per patient for parenteral solutions was $55,193 +/- 30,596 (mean +/- SD) based on Medicare charges and for enteral tube feedings was $9605 +/- 9327. The annual cost of hospitalization ranged from zero to $140,220 in the parenteral nutrition group and from zero to $39,204 in the enteral nutrition group. The annual number of hospitalizations per patient for patients receiving parenteral nutrition ranged from 0.52 to 1.10, compared with 0 to 0.50 in the enteral nutrition population. The health status of HPEN patients was significantly lower (p < .05) in five of the eight short-form 36 health domains compared with the general population. The areas of lifestyle most frequently affected were travel, sleep, exercise and leisure. CONCLUSIONS: The majority of the cost of therapy was associated with the direct provision of nutrition, although in some patients the hospitalization expenditure exceeded this cost. Home nutrition support had a significant negative impact on a patient's quality of life and lifestyle. PMID- 9739035 TI - Pharmacologic levels of heparin do not destabilize neonatal parenteral nutrition. AB - BACKGROUND: Calcium and heparin are known to destabilize the lipid emulsion of total parenteral nutrition (TPN). However, these observations were made over long periods of time, using 5 to 10 times the amount of heparin used in a neonatal intensive care unit. We investigated the effects of lower heparin concentrations with lipid-to-nutrient ratios normally administered to premature infants. METHODS: Lipid emulsion stability was assessed over 30 minutes by measuring viscosity at 10 mmol/L calcium after the addition of 0, 0.5, 1, and 5 IU heparin/mL. This was done at a range of lipid-to-nutrient ratios in clinical use. The effect of varying calcium concentration and different multivitamin preparations on emulsion stability also was observed. RESULTS: Heparin caused an immediate increase in viscosity of pure Intralipid 20% (Intralipid; Kabi Pharmacia AB, Stockholm, Sweden), which eventually separated into two phases. Although changes in viscosity were observed at 1:1 lipid-to-nutrient ratios, no effect was seen at a 1:9 ratio. With the 1:1 ratio, the multivitamin preparations, MVI Pediatric (Rhone-Poulec Rorer, Montreal, Canada) and Vitlipid (Kabi Pharmacia AB), reduced the increase in viscosity. CONCLUSIONS: Heparin and calcium destabilize Intralipid. This is unlikely to cause problems for most infants receiving TPN, provided low heparin concentrations are used. In all cases, especially where the lipid ratio is high, the risk of the lipid phase separating out can be further minimized by (1) the addition of multivitamin preparations containing detergent or an emulsifying agent and (2) by having the shortest possible delivery tube between the point of mixing the lipid and amino acid solutions of TPN and entry into the infant. PMID- 9739036 TI - A randomized, controlled, a single-blind trial of nutritional supplementation after acute stroke. AB - BACKGROUND: Although stroke patients who do not have difficulty swallowing may be at risk of undernutrition and worsening nutritional status during hospitalization, optimum methods for nutrition intervention in stroke patients have not been established. AIM: To examine the feasibility of enteral sip feeding as an effective nutrition intervention after acute stroke. METHODS: Forty-two acute ischemic stroke inpatients with impaired nutritional status who did not have difficulty swallowing within 1 week after the stroke were entered into a single-blind, randomized, controlled, prospective study of enteral sip feeding. Twenty-one patients were randomized to receive daily oral food supplements for 4 weeks in addition to the hospital food, and 21 patients received only the hospital food for the same period. Main outcome measures were energy and protein intakes during the intervention period, change in nutritional status, disability, infective complications, length of stay, and mortality during hospitalization and at 3 months. RESULTS: Two patients, one from each group, were lost to follow-up immediately after randomization. Twenty patients received oral nutritional supplementation. The energy intake was significantly greater in the supplemented group: 1807 +/- 318 vs 1084 +/- 343 kcal/d (mean +/- SD; p < .0001) (estimated treatment effect, 723 kcal/d; 95% confidence interval [CI], 498 to 947), as was protein intake: 65.1 +/- 13.8 vs 44.1 +/- 12.8 g/d (p < .001) (estimated treatment effect, 21.0 g/d; 95% CI, 11.7 to 30.3). There also were significant differences between the two groups in the changes in serum albumin and serum iron concentrations between randomization and at follow-up. There was a trend to lower mortality at 3 months in the supplemented group with two deaths (10%) compared with seven deaths (35%) in the control group (p = .127, relative risk, 0.29; 95% CI, 0.07 to 1.21). CONCLUSIONS: This study suggests that enteral sip feeding is effective in improving nutritional intake and status in stroke patients who do not have swallowing difficulties. There also may be some beneficial effects on clinical outcome, but larger studies are required to confirm this observation and define more precisely the magnitude of any favorable effects. PMID- 9739037 TI - Effects of xylitol on urea synthesis in patients with cirrhosis of the liver. AB - BACKGROUND: In individuals with cirrhosis the normal inhibiting effect of glucose on urea synthesis is lost, probably because of very high concentrations of glucagon. In agreement, glucose does not prevent the inducing effect of glucagon on urea synthesis in normal humans. In contrast, the sugar alcohol, xylitol, prevents the increasing effect of glucagon in normal humans. We, therefore, examined the effect of xylitol on urea synthesis in individuals with cirrhosis and hyperglucagonemia. METHODS: Urea synthesis, calculated as urinary excretion rate corrected for accumulation in total body water and intestinal loss, was measured during infusion of alanine (2 mmol/[h x kg body wt]) and during infusion of alanine superimposed on infusion of xylitol (0.12 g/[h x kg body wt]) in 8 individuals with biopsy-proven alcoholic cirrhosis. The functional hepatic nitrogen clearance (FHNC), ie, urea synthesis expressed independent of changes in plasma amino acid concentration, was calculated as the slope of the linear relation between the urea synthesis rate and the plasma amino acid concentration. RESULTS: All individuals had elevated basal plasma glucagon concentration (261 +/ 61 ng/L; mean +/- SEM) and a markedly increased response to alanine infusion (1037 +/- 226 ng/L). This was not changed by xylitol. Neither the basal urea synthesis rate (13.2 +/- 2.5 mmol/h) nor the alanine-stimulated urea synthesis rate (76.8 +/- 3.64 mmol/h) was changed by xylitol. FHNC during the infusion of alanine alone was 10.5 +/- 0.9 L/h and did not change during the concomitant infusion of xylitol (10.1 +/- 1.1 L/h). CONCLUSIONS: Xylitol reduces neither urea synthesis nor FHNC. The data do not support an important role of xylitol as a nitrogen-sparing agent in cirrhosis. PMID- 9739039 TI - The reliability of muscle function analysis using different methods of stimulation. AB - BACKGROUND: The purpose of our study was to determine the reliability of nonvolitional muscle function analysis (MFA) by determining the day-to-day and within-day reliability of conventional electrical stimulation and a newer, magneto-electrical stimulation method, using standard laboratory methodology. METHODS: Ten healthy, human immunodeficiency virus-negative adult men volunteered as subjects. MFA consisted of measuring the maximal relaxation rate, for magneto electrical stimulation at 1 Hz and conventional electrical stimulation at 20 Hz, and force-frequency ratios using conventional electrical stimulation at 10 Hz:20 Hz and 10 Hz:50 Hz. Within-day and day-to-day reliability were determined by calculating the coefficient of variation (CV) for all subjects. RESULTS: Maximal relaxation rate using magneto-electrical stimulation had a significantly lower CV compared with the other nonvolitional MFA methods (p = .002). CONCLUSIONS: Maximal relaxation rate using magneto-electrical stimulation was more reliable and technically easier than the other muscle function parameters examined. However, the day-to-day CV of muscle function parameters is larger than traditional nutrition assessment techniques. Development within the field should strive to improve testing techniques so that the reliability of MFA will allow definition of a range of normal values against which an individual's value can be compared. Until this is available, the precision and reliability of MFA restrict its use to research and population studies. PMID- 9739038 TI - Epidermal growth factor and human growth hormone induce two sodium-dependent arginine transport systems after massive enterectomy. AB - BACKGROUND: A combination of epidermal growth factor (EGF) and human growth hormone (hGH) after massive enterectomy induces a 400% increase in arginine transport in the remnant distal small intestine. The kinetic mechanism(s) responsible for enhanced arginine transport under these conditions is unknown. METHODS: New Zealand White rabbits underwent 70% midjejunoileal resection. After a 1-week recovery period, animals received hGH (0.2 mg/kg/d IM), EGF (1.5 microg/kg/h SC), hGH + EGF, or vehicle (equal volume) for 7 days. Transport of tritiated arginine into brush border membrane vesicles prepared from distal remnant small intestinal mucosa was quantified in the presence and absence of a sodium gradient over a range of arginine concentrations (25 to 5000 micromol/L). RESULTS: Eadie-Hofstee transformation of the kinetic data demonstrates two sodium dependent arginine transport systems, comprising a high-capacity, low-affinity system and a low-capacity, high-affinity system. A combination of EGF and hGH significantly upregulates both the high-capacity (685%) and low-capacity (350%) maximum transport velocity (Vmax). Additionally, EGF alone significantly upregulates Vmax by 200% in the low-capacity system. There were no significant changes in transport affinity (Km) in either system. CONCLUSIONS: There are two quiescent sodium-dependent arginine transport systems in the distal small intestine. A combination of EGF and hGH after massive enterectomy increase arginine transport by Vmax upregulation in both the high-capacity/low-affinity and low-capacity/high-affinity systems. PMID- 9739040 TI - New surrogate markers for autoimmune disease activity. PMID- 9739041 TI - Mutation of the mouse hepatocyte nuclear factor/forkhead homologue 4 gene results in an absence of cilia and random left-right asymmetry. AB - Winged helix transcription factors play important roles in cellular differentiation and cell-specific gene expression. To define the role of the winged helix factor hepatocyte nuclear factor/forkhead homologue (HFH)-4, a targeted mutation was created in the mouse hfh-4 gene. No expression of HFH-4 was detected in hfh-4(-)/- mice by RNA blot analysis, in situ hybridization, or RT PCR. hfh-4(-)/- mice were noted to have abnormalities of organ situs consistent with random determination of left-right asymmetry. In addition, a complete absence of cilia was noted in hfh-4(-)/- mice. The hfh-4 gene is thus essential for nonrandom determination of left-right asymmetry and development of ciliated cells. Homozygous mutant mice also exhibited prenatal and postnatal growth failure, perinatal lethality and, in some cases, hydrocephalus. RT-PCR revealed an absence of left-right dynein (lrd) expression in the embryonic lungs of hfh-4( )/- mice, suggesting that HFH-4 may act by regulating expression of members of the dynein family of genes. The abnormalities in ciliary development and organ situs in hfh-4(-)/- mice are similar to those observed in human congenital syndromes such as Kartagener syndrome. Targeted mutation of hfh-4 thus provides a model for elucidating the mechanisms regulating ciliary development and determination of left-right asymmetry. PMID- 9739042 TI - A gender-related defect in lipid metabolism and glucose homeostasis in peroxisome proliferator- activated receptor alpha- deficient mice. AB - The peroxisome proliferator-activated receptor alpha (PPARalpha) is a nuclear receptor implicated in the control of cellular lipid utilization. To test the hypothesis that PPARalpha is activated as a component of the cellular lipid homeostatic response, the expression of PPARalpha target genes was characterized in response to a perturbation in cellular lipid oxidative flux caused by pharmacologic inhibition of mitochondrial fatty acid import. Inhibition of fatty acid oxidative flux caused a feedback induction of PPARalpha target genes encoding fatty acid oxidation enzymes in liver and heart. In mice lacking PPARalpha (PPARalpha-/-), inhibition of cellular fatty acid flux caused massive hepatic and cardiac lipid accumulation, hypoglycemia, and death in 100% of male, but only 25% of female PPARalpha-/- mice. The metabolic phenotype of male PPARalpha-/- mice was rescued by a 2-wk pretreatment with beta-estradiol. These results demonstrate a pivotal role for PPARalpha in lipid and glucose homeostasis in vivo and implicate estrogen signaling pathways in the regulation of cardiac and hepatic lipid metabolism. PMID- 9739043 TI - Transgenic expression of the endothelin-B receptor prevents congenital intestinal aganglionosis in a rat model of Hirschsprung disease. AB - The spotting lethal rat, a naturally occurring rodent model of Hirschsprung disease, carries a deletion in the endothelin-B receptor (EDNRB) gene that abrogates expression of functional EDNRB receptors. Rats homozygous for this mutation (sl) exhibit coat color spotting and congenital intestinal aganglionosis. These deficits result from failure of the neural crest-derived epidermal melanoblasts and enteric nervous system (ENS) precursors to completely colonize the skin and intestine, respectively. We demonstrate that during normal rat development, the EDNRB mRNA expression pattern is consistent with expression by ENS precursors throughout gut colonization. We used the human dopamine-beta hydroxylase (DbetaH) promoter to direct transgenic expression of EDNRB to colonizing ENS precursors in the sl/sl rat. The DbetaH-EDNRB transgene compensates for deficient endogenous EDNRB in these rats and prevents the intestinal defect. The transgene has no effect on coat color spotting, indicating the critical time for EDNRB expression in enteric nervous system development begins after separation of the melanocyte lineage from the ENS lineage and their common precursor. The transgene dosage affects both the incidence and severity of the congenital intestinal defect, suggesting dosage-dependent events downstream of EDNRB activation in ENS development. PMID- 9739044 TI - The alpha1 Na,K-ATPase gene is a susceptibility hypertension gene in the Dahl salt-sensitiveHSD rat. AB - Despite the prevalence of essential hypertension, its underlying genetic basis has not been elucidated due to the complexities of its determinants. To identify a hypertension susceptibility gene, we used an approach that integrates molecular, transgenic, and genetic analysis using Dahl salt-sensitive (S) and Dahl salt-resistant (R) rats ascertained for genotype and phenotype. To determine the role of the Dahl S Q276L alpha1 Na,K-ATPase gene variant, we developed transgenic Dahl S rats bearing the Dahl R wild-type (wt) alpha1 Na, K-ATPase cDNA directed by the cognate wt promoter region, Tg[wtalpha1]. Transgenic Dahl S rats exhibited less salt-sensitive hypertension, less hypertensive renal disease, and longer life span when compared with non-transgenic Dahl S controls. Total chromosome 2 linkage analysis of F2(SxR) male rats detects cosegregation of the alpha1 Na,K-ATPase locus with salt-sensitive hypertension. These data support the alpha1 Na,K-ATPase gene as a susceptibility gene for salt-sensitive hypertension in the Dahl S rat model, and provide the basis for the study of the alpha1 Na,K ATPase locus in human hypertension. PMID- 9739046 TI - Coordinated clearance of periciliary liquid and mucus from airway surfaces. AB - Airway surface liquid is comprised of mucus and an underlying, watery periciliary liquid (PCL). In contrast to the well-described axial transport of mucus along airway surfaces via ciliary action, theoretical analyses predict that the PCL is nearly stationary. Conventional and confocal microscopy of fluorescent microspheres and photoactivated fluorescent dyes were used with well differentiated human tracheobronchial epithelial cell cultures exhibiting spontaneous, radial mucociliary transport to study the movements of mucus and PCL. These studies showed that the entire PCL is transported at approximately the same rate as mucus, 39.2+/-4.7 and 39.8+/-4.2 micrometer/sec, respectively. Removing the mucus layer reduced PCL transport by > 80%, to 4.8+/-0.6 micrometer/sec, a value close to that predicted from theoretical analyses of the ciliary beat cycle. Hence, the rapid movement of PCL is dependent upon the transport of mucus. Mucus-dependent PCL transport was spatially uniform and exceeded the rate expected for pure frictional coupling with the overlying mucus layer; hence, ciliary mixing most likely accelerates the diffusion of momentum from mucus into the PCL. The cephalad movement of PCL along airway epithelial surfaces makes this mucus-driven transport an important component of salt and water physiology in the lung in health and disease. PMID- 9739045 TI - CD8 positive T cells influence antigen-specific immune responses through the expression of chemokines. AB - The potential roles of CD8(+) T-cell-induced chemokines in the expansion of immune responses were examined using DNA immunogen constructs as model antigens. We coimmunized cDNA expression cassettes encoding the alpha-chemokines IL-8 and SDF-1alpha and the beta-chemokines MIP-1alpha, RANTES, and MCP-1 along with DNA immunogens and analyzed the resulting antigen-specific immune responses. In a manner more similar to the traditional immune modulatory role of CD4(+) T cells via the expression of Th1 or Th2 cytokines, CD8(+) T cells appeared to play an important role in immune expansion and effector function by producing chemokines. For instance, IL-8 was a strong inducer of CD4(+) T cells, indicated by strong T helper proliferative responses as well as an enhancement of antibody responses. MIP-1alpha had a dramatic effect on antibody responses and modulated the shift of immune responses to a Th2-type response. RANTES coimmunization enhanced the levels of antigen-specific Th1 and cytotoxic T lymphocyte (CTL) responses. Among the chemokines examined, MCP-1 was the most potent activator of CD8(+) CTL activity. The enhanced CTL results are supported by the increased expression of Th1 cytokines IFN-gamma and TNF-alpha and the reduction of IgG1/IgG2a ratio. Our results support that CD8(+) T cells may expand both humoral and cellular responses in vivo through the elaboration of specific chemokines at the peripheral site of infection during the effector stage of the immune response. PMID- 9739047 TI - Circulating, but not local lung, IL-5 is required for the development of antigen induced airways eosinophilia. AB - IL-5 is induced locally in the lung and systemically in the circulation during allergic airways eosinophilic inflammation both in humans and experimental animals. However, the precise role of local and systemic IL-5 in the development of allergic airways eosinophilia remains to be elucidated. In our current study, we demonstrate that compared with their IL-5(+/+) counterparts, IL-5(-/-) mice lacked an IL-5 response both in the lung and peripheral blood, yet they released similar amounts of IL-4, eotaxin, and MIP-1alpha in the lung after ovalbumin (OVA) sensitization and challenge. At cellular levels, these mice failed to develop peripheral blood and airways eosinophilia while the responses of lymphocytes, neutrophils, and macrophages remained similar to those in IL-5(+/+) mice. To dissect the relative role of local and systemic IL-5 in this model, we constructed a gene transfer vector expressing murine IL-5. Intramuscular IL-5 gene transfer to OVA-sensitized IL-5(-/-) mice led to raised levels of IL-5 compartmentalized to the circulation and completely reconstituted airways eosinophilia upon OVA challenge, which was associated with reconstitution of eosinophilia in the bone marrow and peripheral blood. Significant airways eosinophilia was observed for at least 7 d in these mice. In contrast, intranasal IL-5 gene transfer, when rendered to give rise to a significant but compartmentalized level of transgene protein IL-5 in the lung, was unable to reconstitute airways eosinophilia in OVA-sensitized IL-5(-/-) mice upon OVA challenge, which was associated with a lack of eosinophilic responses in bone marrow and peripheral blood. Our findings thus provide unequivocal evidence that circulating but not local lung IL-5 is critically required for the development of allergic airways eosinophilia. These findings also provide the rationale for developing strategies to target circulating IL-5 and/or its receptors in bone marrow to effectively control asthmatic airways eosinophilia. PMID- 9739048 TI - LIGHT, a novel ligand for lymphotoxin beta receptor and TR2/HVEM induces apoptosis and suppresses in vivo tumor formation via gene transfer. AB - LIGHT is a new member of tumor necrosis factor (TNF) cytokine family derived from an activated T cell cDNA library. LIGHT mRNA is highly expressed in splenocytes, activated PBL, CD8(+) tumor infiltrating lymphocytes, granulocytes, and monocytes but not in the thymus and the tumor cells examined. Introduction of LIGHT cDNA into MDA-MB-231 human breast carcinoma caused complete tumor suppression in vivo. Histological examination showed marked neutrophil infiltration and necrosis in LIGHT expressing but not in the parental or the Neo-transfected MDA-MB-231 tumors. Interferon gamma (IFNgamma) dramatically enhances LIGHT-mediated apoptosis. LIGHT protein triggers apoptosis of various tumor cells expressing both lymphotoxin beta receptor (LTbetaR) and TR2/HVEM receptors, and its cytotoxicity can be blocked specifically by addition of a LTbetaR-Fc or a TR2/HVEM-Fc fusion protein. However, LIGHT was not cytolytic to the tumor cells that express only the LTbetaR or the TR2/HVEM or hematopoietic cells examined that express only the TR2/HVEM, such as PBL, Jurkat cells, or CD8(+) TIL cells. In contrast, treatment of the activated PBL with LIGHT resulted in release of IFNgamma. Our data suggest that LIGHT triggers distinct biological responses based on the expression patterns of its receptors on the target cells. Thus, LIGHT may play a role in the immune modulation and have a potential value in cancer therapy. PMID- 9739049 TI - Generation of lyso-phospholipids from surfactant in acute lung injury is mediated by type-II phospholipase A2 and inhibited by a direct surfactant protein A phospholipase A2 protein interaction. AB - Lyso-phospholipids exert a major injurious effect on lung cell membranes during Acute Respiratory Distress Syndrome (ARDS), but the mechanisms leading to their in vivo generation are still unknown. Intratracheal administration of LPS to guinea pigs induced the secretion of type II secretory phospholipase A2 (sPLA2 II) accompanied by a marked increase in fatty acid and lyso-phosphatidylcholine (lyso-PC) levels in the bronchoalveolar lavage fluid (BALF). Administration of LY311727, a specific sPLA2-II inhibitor, reduced by 60% the mass of free fatty acid and lyso-PC content in BALF. Gas chromatography/mass spectrometry analysis revealed that palmitic acid and palmitoyl-2-lyso-PC were the predominant lipid derivatives released in BALF. A similar pattern was observed after the intratracheal administration of recombinant guinea pig (r-GP) sPLA2-II and was accompanied by a 50-60% loss of surfactant phospholipid content, suggesting that surfactant is a major lung target of sPLA2-II. In confirmation, r-GP sPLA2-II was able to hydrolyze surfactant phospholipids in vitro. This hydrolysis was inhibited by surfactant protein A (SP-A) through a direct and selective protein protein interaction between SP-A and sPLA2-II. Hence, our study reports an in vivo direct causal relationship between sPLA2-II and early surfactant degradation and a new process of regulation for sPLA2-II activity. Anti-sPLA2-II strategy may represent a novel therapeutic approach in lung injury, such as ARDS. PMID- 9739050 TI - Activated endothelial cells elicit paracrine induction of epithelial chloride secretion. 6-Keto-PGF1alpha is an epithelial secretagogue. AB - Endothelial cells play a central role in the coordination of the inflammatory response. In mucosal tissue, such as the lung and intestine, endothelia are anatomically positioned in close proximity to epithelia, providing the potential for cell-cell crosstalk. Thus, in this study endothelial-epithelial biochemical crosstalk pathways were studied using a human intestinal crypt cell line (T84) grown in noncontact coculture with human umbilical vein endothelia. Exposure of such cocultures to endothelial-specific agonists (LPS) resulted in activation of epithelial electrogenic Cl- secretion and vectorial fluid transport. Subsequent experiments revealed that in response to diverse stimuli (LPS, IL-1alpha, TNF alpha, hypoxia), endothelia produce and secrete a small, stable epithelial secretagogue into conditioned media supernatants. Further experiments identified this secretagogue as 6-keto-PGF1alpha, a stable hydrolysis product of prostacyclin (PGI2). Results obtained with synthetic prostanoids indicated that 6 keto-PGF1alpha (EC50 = 80 nM) and PGI2 stable analogues (EC50 = 280 nM) activate the same basolaterally polarized, Ca2+-coupled epithelial receptor. In summary, these findings reveal a previously unappreciated 6-keto-PGF1alpha receptor on intestinal epithelia, the ligation of which results in activation of electrogenic Cl- secretion. In addition, these data reveal a novel action for the prostacyclin hydrolysis product 6-keto-PGF1alpha and provide a potential endothelial- epithelial crosstalk pathway in mucosal tissue. PMID- 9739051 TI - Functional activation of lymphocyte CD44 in peripheral blood is a marker of autoimmune disease activity. AB - Interactions between complementary receptors on leukocytes and endothelial cells play a central role in regulating extravasation from the blood and thereby affect both normal and pathologic inflammatory responses. CD44 on lymphocytes that has been "activated" to bind its principal ligand hyaluronate (HA) on endothelium can mediate the primary adhesion (rolling) of lymphocytes to vascular endothelial cells under conditions of physiologic shear stress, and this interaction is used for activated T cell extravasation into an inflamed site in vivo in mice (DeGrendele, H.C., P. Estess, L.J. Picker, and M.H. Siegelman. 1996. J. Exp. Med. 183:1119-1130. DeGrendele, H.D., P. Estess, and M.H. Siegelman. 1997. Science. 278:672-675. DeGrendele, H.C., P. Estess, and M.H. Siegelman. 1997. J. Immunol. 159: 2549-2553). Here, we have investigated the role of lymphocyte-borne activated CD44 in the human and show that CD44-dependent primary adhesion is induced in human peripheral blood T cells through T cell receptor triggering. In addition, lymphocytes capable of CD44/HA-dependent rolling interactions can be found resident within inflamed tonsils. In analysis of peripheral bloods of patients from a pediatric rheumatology clinic, examining systemic lupus erythematosus, and a group of chronic arthropathies, expression of CD44-dependent primary adhesion strongly correlates with concurrent symptomatic disease, with 85% of samples from clinically active patients showing elevated levels of rolling activity (compared with only 4% of inactive patients). These rolling interactions are predominantly mediated by T cells. The results suggest that circulating T lymphocytes bearing activated CD44 are elevated under conditions of chronic inflammation and that these may represent a pathogenically important subpopulation of activated circulating cells that may provide a reliable marker for autoimmune or chronic inflammatory disease activity. PMID- 9739052 TI - A two-amino acid insertion in the Cys146- Cys167 loop of the alphaIIb subunit is associated with a variant of Glanzmann thrombasthenia. Critical role of Asp163 in ligand binding. AB - The ligand binding site(s) of the alpha subunit of integrin alphaIIb beta3 (GPIIb IIIa), a prototypic non-I domain integrin, remains elusive. In this study, we have characterized a Japanese variant of Glanzmann thrombasthenia, KO, whose platelets express normal amounts of alphaIIb beta3. KO platelets failed to bind the activation-independent ligand-mimetic mAb OP-G2 and did not bind fibrinogen or the activation-dependent ligand-mimetic mAb PAC-1 following activation of alphaIIb beta3 under any condition examined. Sequence analysis of PCR fragments derived from KO platelet mRNA revealed a 6-bp insertion leading to a 2-amino-acid insertion (Arg-Thr) between residues 160 and 161 of the alphaIIb subunit. Introduction of the insertion into wild-type recombinant alphaIIb beta3 expressed in 293 cells led to the normal expression of alphaIIb beta3 having the defect in ligand binding function. The insertion is located within the small loop (Cys146 Cys167) in the third NH2-terminal repeat of the alphaIIb subunit. Alanine substitution of each of the oxygenated residues within the loop (Thr150, Ser152, Glu157, Asp159, Ser161, and Asp163) did not significantly affect expression of alphaIIbbeta3, and only Asp163AlaalphaIIb beta3 abolished the ligand binding function. In addition, Asp163AlaalphaIIb beta3 as well as KO mutant alphaIIb beta3 constitutively expressed the PMI-1 epitope. Our present data suggest that Asp163 of the alphaIIb subunit is one of the critical residues for ligand binding. PMID- 9739053 TI - Mild trifunctional protein deficiency is associated with progressive neuropathy and myopathy and suggests a novel genotype-phenotype correlation. AB - Human mitochondrial trifunctional protein (TFP) is a heterooctamer of four alpha- and four beta-subunits that catalyzes three steps in the beta-oxidation spiral of long-chain fatty acids. TFP deficiency causes a Reye-like syndrome, cardiomyopathy, or sudden, unexpected death. We delineated the molecular basis for TFP deficiency in two patients with a unique phenotype characterized by chronic progressive polyneuropathy and myopathy without hepatic or cardiac involvement. Single-stranded conformation variance and nucleotide sequencing identified all patient mutations in exon 9 of the alpha-subunit. One patient is homozygous for the T845A mutation that substitutes aspartic acid for valine at residue 246. The second patient is a compound heterozygote for the T914A that substitutes asparagine for isoleucine at residue 269 and a C871T that creates a premature termination at residue 255. Allele-specific oligonucleotide hybridization studies revealed undetectable levels of the mRNA corresponding to the mutant allele carrying the termination codon. This study suggests a novel genotype-phenotype correlation in TFP deficiency; that is, mutations in exon 9 of the alpha-subunit, which encodes a linker domain between the NH2-terminal hydratase and the COOH-terminal 3-hydroxyacyl-CoA dehydrogenase, result in a unique neuromuscular phenotype. PMID- 9739055 TI - An endothelial growth factor involved in rat renal development. AB - In the kidney, there is a close and intricate association between epithelial and endothelial cells, suggesting that a complex reciprocal interaction may exist between these two cell types during renal ontogeny. Thus, we examined whether metanephrogenic mesenchymal cells secrete endothelial mitogens. With an endothelial mitogenic assay and sequential chromatography of the proteins in the media conditioned by a cell line of rat metanephrogenic mesenchymal cells (7.1.1 cells), we isolated a protein whose amino acid analysis identified it as hepatoma derived growth factor (HDGF). Media conditioned with Cos-7 cell transfected with HDGF cDNA stimulated endothelial DNA synthesis. With immunoaffinity purified antipeptide antibodies, we found that HDGF was widely distributed in the renal anlage at early stages of development but soon concentrated at sites of active morphogenesis and, except for some renal tubules, disappeared from the adult kidney. From a 7.1.1 cells cDNA library, a clone of most of the translatable region of HDGF was obtained and used to synthesize digoxigenin-labeled riboprobes. In situ hybridization showed that during kidney development mRNA for HDGF was most abundant at sites of nephron morphogenesis and in ureteric bud cells while in the adult kidney transcripts disappeared except for a small population of distal tubules. Thus, HDGF is an endothelial mitogen that is present in embryonic kidney, and its expression is synchronous with nephrogenesis. PMID- 9739054 TI - Inducible nitric oxide synthase expression is reduced in cystic fibrosis murine and human airway epithelial cells. AB - It has been reported that exhaled nitric oxide levels are reduced in cystic fibrosis (CF) patients. We have examined the inducible isoform of nitric oxide synthase (iNOS) in the airways by immunostaining and found that iNOS is constitutively expressed in the airway epithelia of non-CF mouse and human tissues but essentially absent in the epithelium of CF airways. We explored potential consequences of lost iNOS expression and found that iNOS inhibition significantly increases mouse nasal trans-epithelial potential difference, and hindered the ability of excised mouse lungs to prevent growth of Pseudomonas aeruginosa. The absence of continuous nitric oxide production in epithelial cells of CF airways may play a role in two CF-associated characteristics: hyperabsorption of sodium and susceptibility to bacterial infections. PMID- 9739056 TI - Protective role of endogenous carbon monoxide in hepatic microcirculatory dysfunction after hemorrhagic shock in rats. AB - Maintenance of hepatic microcirculatory flow after ischemia of the liver is essential to prevent hepatic dysfunction. Thus, we determined the differential role of carbon monoxide (CO) and nitric oxide (NO) in the intrinsic control of sinusoidal perfusion, mitochondrial redox state, and bile production in the isolated perfused rat liver after hemorrhagic shock. Administration of tin protoporphyrin-IX (50 microM), a specific inhibitor of the CO generating enzyme heme oxygenase, caused a decrease in sinusoidal flow that was more pronounced after shock compared with sham shock, as determined by in situ epifluorescence microscopy. This was associated with a shift in hepatocellular redox potential to a more reduced state (increased fluorescence intensity of reduced pyridine nucleotides in hepatocytes, decreased acetoacetate/beta-hydroxybutyrate ratio in the perfusate) and a profound reduction in bile flow. In sharp contrast, the preferential inhibitor of the inducible isoform of NO synthase S methylisothiourea sulfate (100 microM) did not affect sinusoidal flow, hepatic redox state, or function. This indicates that 1.) endogenously generated CO preserves sinusoidal perfusion after hemorrhagic shock, 2.) protection of the hepatic microcirculation by CO may serve to limit shock-induced liver dysfunction, and 3.) in contrast to CO, inducible NO synthase-derived NO is of only minor importance for the intrinsic control of hepatic perfusion and function under these conditions. PMID- 9739057 TI - Resistance of Fc receptor- deficient mice to fatal glomerulonephritis. AB - Immune complex-mediated inflammation is a common mechanism of various autoimmune diseases. Glomerulonephritis (GN) is one of these diseases, and the main mechanism of the induction of GN has been unclear. We examined the contribution of Fc receptors in the induction of nephrotoxic GN by establishing and analyzing mice deficient in the Fc receptor gamma chain (FcRgamma). Whereas all wild-type mice died from severe glomerulonephritis with hypernitremia by administration of anti-glomerular basement membrane (GBM) antibodies, all FcRgamma-deficient mice survived. Histologically, wild-type mice showed glomerular hypercellularity and thrombotic changes, whereas the renal tissue in FcRgamma-deficient mice was almost intact. Deposition of anti-GBM antibody as well as complement components in the GBM were equally observed in both wild-type and knockout mice. These results demonstrate that the triggering of this type of glomerulonephritis is completely dependent on FcR+ cells. PMID- 9739058 TI - Enhanced in vitro potency and in vivo immunogenicity of a CTL epitope from hepatitis C virus core protein following amino acid replacement at secondary HLA A2.1 binding positions. AB - Since the natural immune response to hepatitis C virus (HCV) is often unable to clear the infection, to enhance immunogenicity we studied substituted peptides from an HCV cytotoxic T lymphocyte (CTL) epitope (C7A2) from a conserved region of the HCV core protein (DLMGYIPLV) recognized by CTL lines from HLA-A2.1(+) HCV infected patients and HLA-A2.1 transgenic mice. HLA-A2.1 binding, human and murine CTL recognition, and in vivo immunogenicity (using mice transgenic for human HLA-A2 in lieu of immunizing humans) were analyzed to define peptides with enhanced immunogenicity. Peptides substituted at position 1 showed enhanced HLA A2 binding affinity, but paradoxically poorer immunogenicity. A peptide with Ala substituted at position 8 (8A) showed higher HLA-A2 binding affinity and CTL recognition and was a more potent in vivo immunogen in HLA-A2-transgenic mice, inducing higher CTL responses with higher avidity against native C7A2 than induced by C7A2 itself. These results suggest that peptide 8A is a more potent in vitro antigen and in vivo immunogen than C7A2 and may be useful as a vaccine component. They provide proof of principle that the strategy of epitope enhancement can enhance immunogenicity of a CTL epitope recognized by human CTL. PMID- 9739059 TI - IFN-gamma action on pancreatic beta cells causes class I MHC upregulation but not diabetes. AB - We have generated transgenic nonobese diabetic (NOD) mice expressing dominant negative mutant IFN-gamma receptors on pancreatic beta cells to investigate whether the direct effects of IFN-gamma on beta cells contribute to autoimmune diabetes. We have also quantitated by flow cytometry the rise in class I MHC on beta cells of NOD mice with increasing age and degree of islet inflammatory infiltrate. Class I MHC expression increases gradually with age in wild-type NOD mice; however, no such increase is observed in the transgenic beta cells. The transgenic mice develop diabetes at a similar rate to that of wild-type animals. This study dissociates class I MHC upregulation from progression to diabetes, shows that the rise in class I MHC is due to local IFN-gamma action, and eliminates beta cells as the targets of IFN-gamma in autoimmune diabetes. PMID- 9739062 TI - Review: Evolutionary link between prokaryotic and eukaryotic K+ channels. AB - Considering the importance of K+ channels in controlling the crucial K+ gradient across the plasma membranes of all living cells, it comes as no surprise that, besides being present in every eukaryotic cell, these integral membrane proteins have recently also been identified in prokaryotes. Today, approximately a dozen successfully completed and many more ongoing sequencing projects permit a search for genes related to K+ channels in the genomes of both eubacteria and archaea. The coding regions of homologues show a remarkable variety in primary structure. They predict membrane proteins with one, two, three and six hydrophobic segments surrounding a putative K+-selective pore (H5) and the presence or absence of a cytosolic putative NAD+-binding domain (PNBD) that probably senses the reducing power of the cell. The analysis of kinships on the basis of phylogenetic algorithms identifies sequences closely related to eukaryotic voltage-dependent Kv channels, but also defines members of a primordial class of prokaryotic K+ channel (containing the 2TMS/PNBD motif). Considering the unique mechanisms that may account for the assembly of modern proteins from different ancestral genes, and with more primary sequence data soon to appear, a scheme for the evolutionary origin of K+ channels comes within reach. PMID- 9739061 TI - MHC haplotype-dependent regulation of MOG-induced EAE in rats. AB - Experimental autoimmune encephalomyelitis (EAE) induced in the rat by active immunization with myelin-oligodendrocyte-glycoprotein (MOG) is mediated by synergy between MOG-specific T cells and demyelinating MOG-specific antibody responses. The resulting disease is chronic and displays demyelinating central nervous system (CNS) pathology that closely resembles multiple sclerosis. We analyzed major histocompatibility complex (MHC) haplotype influences on this disease. The MHC haplotype does not exert an all-or-none effect on disease susceptibility. Rather, it determines the degree of disease susceptibility, recruitment of MOG-specific immunocompetent cells, clinical course, and CNS pathology in a hierarchical and allele-specific manner. Major haplotype-specific effects on MOG-EAE map to the MHC class II gene region, but this effect is modified by other MHC genes. In addition, non-MHC genes directly influence both disease and T cell functions, such as the secretion of IFN-gamma. Thus, in MOG EAE, allelic MHC class II effects are graded, strongly modified by other MHC genes, and overcome by effects of non-MHC genes and environment. PMID- 9739063 TI - Molecular cloning and developmental expression patterns of the MyoD and MEF2 families of muscle transcription factors in the carp. AB - cDNA clones encoding the myogenic regulatory factors (MRFs) myogenin, MyoD and myf-5 were isolated by reverse-transcription polymerase chain reaction from larvae and embryos of the common carp (Cyprinus carpio L.). Myocyte-specific enhancer factor 2 (MEF2) cDNAs were identified from a cDNA library from adult carp. Northern blot analysis showed that MyoD, myf-5 and MEF2C transcripts were present in three-somite embryos, whereas myogenin and MEF2A transcripts were not detected until the 15-somite stage. Intense signals of myogenin and MyoD transcripts were observed even in 1-month-old juveniles. Levels of MyoD, myogenin and MEF2A transcripts declined between 1 and 7 months after hatching, and myf-5 gave only a weak signal in the oldest fish. In contrast, levels of MEF2C transcripts were considerably higher in 7-month-old juveniles than in 1-month-old larvae. mRNAs encoding carp myosin heavy chain and -actin were first detected at approximately the time of the first heartbeat, and levels were maximal in juveniles 1 month post-hatching. The relatively high levels of MRF mRNA in juvenile fish probably reflect the recruitment of new muscle fibres from the satellite cell population. It was concluded that the relative importance of the different members of the MyoD and MEF2 families of transcription factors for muscle differentiation changes during ontogeny in the carp. PMID- 9739064 TI - Two types of mRNA encoding myosin regulatory light chain in carp fast skeletal muscle differ in their 3' non-coding regions and expression patterns following temperature acclimation. AB - cDNA clones encoding the myosin regulatory light chain (RLC) were isolated from a cDNA library prepared from fast skeletal muscle of the carp Cyprinus carpio L. Two types of cDNA clone encoding carp RLC were found with identical deduced amino acid sequences. The two mRNAs differed in the number of polyadenylation signals prior to the poly(A) tail in the 3' non-coding region. The two mRNA species, with approximate sizes of 1.4 and 0.8 kilobases, were also observed in northern blot analysis. Carp were acclimated for a minimum of 5 weeks to either 10 degreesC or 30 degreesC (14 h:10 h light:dark photoperiod). The total levels of mRNA transcripts coding for the RLC and myosin heavy chain were, respectively, 3.3 and 3.9 times higher in cold- than in warm-acclimated fish. Differences in the levels of RLC in mRNA transcripts were largely due to the concentration of the 1.4 kilobase mRNA species. PMID- 9739060 TI - Tumor immunity and autoimmunity induced by immunization with homologous DNA. AB - The immune system can recognize self antigens expressed by cancer cells. Differentiation antigens are prototypes of these self antigens, being expressed by cancer cells and their normal cell counterparts. The tyrosinase family proteins are well characterized differentiation antigens recognized by antibodies and T cells of patients with melanoma. However, immune tolerance may prevent immunity directed against these antigens. Immunity to the brown locus protein, gp75/ tyrosinase-related protein-1, was investigated in a syngeneic mouse model. C57BL/6 mice, which are tolerant to gp75, generated autoantibodies against gp75 after immunization with DNA encoding human gp75 but not syngeneic mouse gp75. Priming with human gp75 DNA broke tolerance to mouse gp75. Immunity against mouse gp75 provided significant tumor protection. Manifestations of autoimmunity were observed, characterized by coat depigmentation. Rejection of tumor challenge required CD4(+) and NK1.1(+) cells and Fc receptor gamma-chain, but depigmentation did not require these components. Thus, immunization with homologous DNA broke tolerance against mouse gp75, possibly by providing help from CD4(+) T cells. Mechanisms required for tumor protection were not necessary for autoimmunity, demonstrating that tumor immunity can be uncoupled from autoimmune manifestations. PMID- 9739065 TI - Morphological, physiological and metabolic comparisons between runner-like and sheet-like inbred lines of a colonial hydroid. AB - Hydractiniid hydroids display a range of morphological variation from sheet-like forms (i.e. closely spaced polyps with high rates of stolon branching) to runner like forms (i.e. widely spaced polyps with low rates of stolon branching), thus exemplifying the patterns of heterochrony found in many colonial animals. A sheet like and a runner-like inbred line of Podocoryne carnea were produced to investigate this heterochronic variation further. Selection on colony morphology at the time of the initiation of medusa production resulted in dramatic differences by the F5 and F6 generations. Compared with colonies of the sheet like inbred line, runner-like colonies exhibited smaller sizes at the initiation of medusa production, more irregular colony shapes and diminished stolon development relative to polyp development. In addition to these differences in colony morphology, runner-like colonies also exhibited larger medusae and a greater amount of gastrovascular flow to the peripheral stolons. To assess differences in the metabolic capacity underlying this variaton in flow, the redox state of the polyp epitheliomuscular cells was measured using the fluorescence of NAD(P)H. In response to feeding-induced changes in gastrovascular flow, runner like colonies show greater redox variation than sheet-like ones, plausibly corresponding to the greater amounts of flow generated by the former colonies relative to the latter. Perturbing the system with dilute solutions of 2,4 dinitrophenol similarly indicates that runner-like colonies contain more functionally oxidizable NAD(P)H. The correlation between gastrovascular flow and morphological differences supports the hypothesis that the former mediates the timing of colony development, perhaps in concert with the observed variation in the redox state of polyp epitheliomuscular cells. PMID- 9739066 TI - Glutamatergic neuromuscular transmission in the heart of the isopod crustacean Ligia exotica. AB - Neuromuscular transmission between the cardiac ganglion (CG) and the myocardium was examined in the adult heart of the isopod crustacean Ligia exotica. Intracellular injection of neurobiotin into the CG neurones revealed that all six CG neurones send their axons onto the cardiac muscle, where they form axon terminals bearing varicosities. All the CG neurones and their processes exhibited glutamate-like immunoreactivity. The cardiac muscle showed depolarizing membrane potential responses to glutamate applied focally to sites close to axon terminals bearing varicosities. Both the glutamate-induced response and the excitatory junctional potential (EJP) showed desensitization in response to the repeated application of glutamate. Under voltage-clamp conditions, the cardiac muscle produced inward current responses to focally applied glutamate. The reversal potential for the glutamate-induced current estimated from extrapolation of the linear current/voltage relationship was similar to that of the excitatory junctional current evoked by ganglionic nerve stimulation. Both the glutamate induced response and the EJP were blocked by a glutamate-specific antagonist, Joro spider toxin. These results led us to conclude that the CG neurones of Ligia exotica are glutamatergic motoneurones. PMID- 9739067 TI - Acid-base and respiratory responses to hypoxia in the grasshopper Schistocerca americana. AB - How do quiescent insects maintain constant rates of oxygen consumption at ambient PO2 values as low as 2-5 kPa? To address this question, we examined the response of the American locust Schistocerca americana to hypoxia by measuring the effect of decreasing ambient PO2 on haemolymph acid-base status, tracheal PCO2 and CO2 emission. We also tested the effect of hypoxia on convective ventilation using a new optical technique which measured the changes in abdominal volume during ventilation. Hypoxia caused a progressive increase in haemolymph pH and a decrease in haemolymph PCO2. A Davenport analysis suggests that hypoxia is accompanied by a net transfer of base to the haemolymph, perhaps as a result of intracellular pH regulation. Hypoxia caused a progressive increase in convective ventilation which was mostly attributable to a rise in ventilatory frequency. Carbon dioxide conductance ( micromol h-1 kPa-1) across the spiracles increased more than threefold, while conductance between the haemolymph and primary trachea nearly doubled in 2 kPa O2 relative to room air. The rise in trans-spiracular conductance is completely attributable to the elevations in convective ventilation. The rise in tracheal conductance in response to hypoxia may reflect the removal of fluid from the tracheoles described by Wigglesworth. The low critical PO2 of quiescent insects can be attributed (1) to their relatively low resting metabolic rates, (2) to the possession of tracheal systems adapted for the exchange of gases at much higher rates during activity and (3) to the ability of insects to rapidly modulate tracheal conductance. PMID- 9739068 TI - Functional partitioning of energy reserves by larvae of the marine bryozoan Bugula neritina (L.). AB - The effects of extended swimming on short-lived lecithotrophic larvae of the marine bryozoan Bugula neritina (L.) were examined. Larvae were forced to swim for 2 or 24 h by bath application of serotonin. Settlement and metamorphosis success were significantly reduced, larval dimensions were unaffected and ancestrulae were smaller after 24 h of swimming. Larvae settled predominantly on seawater-conditioned glass after 2 h, but became less discriminative after 24 h. Lipid content in intact larvae and dissociated surface ciliated and interior cell fractions was analysed by thin-layer chromatography. Hydrophilic lipids were unaffected by swimming regime. The hydrophobic fraction contained triglyceride, confirmed by proton nuclear magnetic resonance spectroscopy (1H-NMR) analysis and correlation spectroscopy (1H-1H COSY) patterns, which was significantly depleted after 24 h, and diacylglycerol, which was not. NMR spectra suggested no differences in fatty acid chain compositions between larvae swimming for 2 and 24 h. Triglyceride depletion was limited to the ciliated cell fraction. We propose that the functional partitioning of lipid reserves has evolved in association with the costs and benefits linked with larval dispersal. PMID- 9739069 TI - Comparative kinematics and hydrodynamics of odontocete cetaceans: morphological and ecological correlates with swimming performance. AB - Propulsive morphology and swimming performance were compared for the odontocete cetaceans Delphinapterus leucas, Orcinus orca, Pseudorca crassidens and Tursiops truncatus. Morphological differences were apparent among the whales. The general body contour and low-aspect-ratio caudal flukes of D. leucas indicated that this species was a low-performance swimmer compared with the other species. Propulsive motions were video-taped as animals swam steadily in large pools. Video tapes were analyzed digitally using a computerized motion-analysis system. Animals swam at relative velocities ranging from 0.4 to 2.4 body lengths s-1. The stroke amplitude of the flukes decreased linearly with velocity for D. leucas, but amplitude remained constant for the other species. Tail-beat frequencies were directly related to relative swimming velocity, whereas the pitch angle of the flukes was inversely related to relative swimming velocity. Unsteady lifting-wing theory was used with regression equations based on kinematics to calculate thrust power output, drag coefficients and propulsive efficiency. Compared with other species, O. orca generated the largest thrust power (36.3 kW) and had the lowest drag coefficient (0.0026), whereas T. truncatus displayed the largest mass specific thrust power (23.7 W kg-1) and P. crassidens had the highest efficiency (0.9). D. leucas did not swim as rapidly as the other species and had a comparatively higher minimum drag coefficient (0.01), lower mass-specific thrust power (5.2 W kg-1) and lower maximum efficiency (0.84). Minimum drag coefficients were associated with high swimming speeds, and maximum efficiencies corresponded with velocities in the range of typical cruising speeds. The results indicate that the kinematics of the propulsive flukes and hydrodynamics are associated with the swimming behaviors and morphological designs exhibited by the whales in this study, although additional factors will influence morphology. PMID- 9739070 TI - Receptor cell habituation in the A1 auditory receptor of four noctuoid moths. AB - Moths of both sexes of Empyreuma affinis (=pugione) and Syntomeida epilais (Arctiidae, Ctenuchinae), Maenas jussiae (Arctiidae, Arctiinae) and Spodoptera frugiperda (Noctuidae, Amphipyrinae) were studied. Spike activity in the A1 cell was recorded using a stainless-steel hook electrode from the tympanic nerve in the mesothorax. Acoustic stimuli consisting of 25 and 100 ms pulses at the best frequency for the species and at intensities that evoke A1 cell saturation response were used at repetition rates of 0.5 and 5 Hz for 100 ms stimuli, and between 2 and 20 Hz for 25 ms stimuli. Stimuli at a repetition rate corresponding to a duty cycle of 5 % (25 ms at 2 Hz and 100 ms at 0.5 Hz) did not evoke monotonic changes in the responses of the A1 cell. With 25 ms pulses, rates above 5 Hz evoked an exponential decrease in the number of spikes and an increase in the latency of the responses of all the 37 specimens tested. The response duration showed no apparent change with stimulus repetition rates even at the highest duty cycle used (50 %), i.e. 25 ms at 20 Hz and 100 ms at 5 Hz. The higher the rate of stimulus repetition, the more marked were the changes in the A1 cell responses. In 16 of 17 preparations from two species, habituation had no effect on the adaptation rate in each response, while in seven of eight specimens of another species, the adaptation rate decreased with stimulus repetition. These results, and those from another mechanoreceptor cell, indicate that receptor cell adaptation (changes evoked in the response by a stimulus of constant intensity) and habituation (changes in the responses due to stimulus repetition rate) are two distinctive phenomena. The A1 cell in its habituated state showed an increase in its response to incremental increases in stimulus intensity of 10 dB. This result supports the idea that receptor cell habituation does not seem to be due to fatigue, i.e. to a temporary loss of the ability to respond to stimulation induced in a sensory receptor by continued stimulation. PMID- 9739071 TI - Mechanics of lung ventilation in a larval salamander Ambystoma tigrinum. AB - The larval stage of the tiger salamander Ambystoma tigrinum is entirely aquatic, but the larvae rely on their lungs for a large proportion of their oxygen uptake. X-ray video and pressure measurements from the buccal and body cavities demonstrate that the larvae inspire using a two-stroke buccal pump and exhale actively by contracting the hypaxial musculature to increase body pressure. Larvae begin a breath by expanding the buccal cavity to draw in air through the mouth, while simultaneously exhaling air from the lungs to mix with the fresh air in the buccal cavity. The mouth then closes, and the buccal cavity compresses to pump a portion of the mixture into the lungs. The remaining air in the buccal cavity is then released as bubbles from the mouth and gill slits. Ventilatory volumes estimated from X-ray video records indicate that approximately 80 % of the air pumped into the lungs is fresh air and 20 % is previously expired air. Exhalation in larval tiger salamanders is active, powered by contraction of all four layers of lateral hypaxial musculature. Electromyography indicates that the transverse abdominis (TA) muscle is active for the longest duration and shows the highest-amplitude activity, but the external oblique superficialis, the external oblique profundus and the internal oblique also show consistent, low-level activity. The finding that the TA muscle is active during exhalation in larval tiger salamanders contributes to a growing body of evidence that the use of the TA for exhalation is a primitive character for tetrapods. PMID- 9739072 TI - Diet quality influences the (&dgr ;)13C and (&dgr ;)15N of locusts and their biochemical components. AB - To determine whether relative enrichments of 15N and 13C in locusts are influenced by diet, locust nymphs were raised from hatchlings to adults on either seedling wheat or maize. Maize provided less hexose sugars and protein per gram than did wheat. Maize also depends on the C4 form of photosynthesis, while wheat uses the C3 form; this difference in photosynthetic pathways produces two distinguishable ranges of 13C values.The lower-quality maize diet corresponded to a 5.1 increase in animal 15N, relative to diet, whereas the wheat diet corresponded to an increase of only 2.3 . The maize-fed animals were more 13C depleted in lipid, trehalose and chitin than those fed wheat. The results for 15N and 13C suggest that substrate recycling occurred on the low-quality maize diet. Consequently, we examined the variations in the isotopic differences between locusts and their diet at the biochemical level. PMID- 9739073 TI - Intracellular trafficking of the vacuolar H+-ATPase accessory subunit Ac45. AB - Ac45 is a type I transmembrane protein associated with vacuolar H+-ATPase, a proton pump mediating the acidification of multiple intracellular organelles. In this study, we examined the intracellular routing of Ac45 in transfected CV-1 fibroblasts. Steady state immunolabeling showed that Ac45 is located on the plasma membrane and in a vacuolar compartment in the juxtanuclear region. Antibody internalization experiments revealed that Ac45 is rapidly retrieved from the cell surface and is targeted to the vacuolar structures. The 26-residue cytoplasmic tail of Ac45 was intrinsically capable of mediating endocytosis of the cell surface protein Tac, indicating that the tail contains an autonomous internalization signal. Immunolocalization studies on cells expressing carboxy terminally truncated Ac45 mutants showed the presence of essential routing information in the membrane-distal region of the cytoplasmic tail. Further mutational analysis of this region, which lacks the recognized tyrosine- or di leucine-based sorting motifs, suggested that multiple sites rather than a short linear sequence are responsible for the internalization. Collectively, our results indicate that the cytoplasmic tail of Ac45 contains autonomous targeting information distinct from previously described routing determinants. PMID- 9739074 TI - Polarized localizations of annexins I, II, VI and XIII in epithelial cells of intestinal, hepatic and pancreatic tissues. AB - The cellular and subcellular localizations of annexins I, II, VI and XIII in the rabbit intestine, liver and pancreas were studied by performing immunofluorescence labeling on thin frozen tissue sections using specific monoclonal antibodies. The expression of annexins was found to be finely regulated. Annexins XIII and I were expressed exclusively in the small intestine and the colon, respectively, whereas annexin II was present in all the tissues tested and annexin VI specifically in the liver and pancreas. These different annexins were concentrated in the basolateral domain of polarized cells, and some of them had an extra-apical localization: annexin XIII was concentrated in the lower 3/4 of enterocyte brush border microvilli; annexin II was present in the upper part of the terminal web in intestinal absorbent cells as well as in the bile canalicular area in hepatocytes, whereas annexin VI was detected on some apical vesicles concentrated around the bile canaliculi. In pancreatic acinar cells, the presence of annexin II on some zymogen granules provides further evidence that annexin II may be involved in exocytic events. In conclusion, this study shows that the basolateral domain of polarized cells appears to be the main site where annexins are located, and they may therefore be involved in the important cellular events occurring at this level. PMID- 9739075 TI - The role of the ran GTPase in nuclear assembly and DNA replication: characterisation of the effects of Ran mutants. AB - The Ran GTPase plays a critical role in nucleocytoplasmic transport and has been implicated in the maintenance of nuclear structure and cell cycle control. Here, we have investigated its role in nuclear assembly and DNA replication using recombinant wild-type and mutant Ran proteins added to a cell-free system of Xenopus egg extracts. RanQ69L and RanT24N prevent lamina assembly, PCNA accumulation and DNA replication. These effects may be due to the disruption of nucleocytoplasmic transport, since both mutants inhibit nuclear import of a protein carrying a nuclear localisation signal (NLS). RanQ69L, which is deficient in GTPase activity, sequesters importins in stable complexes that are unable to support the docking of NLS-proteins at the nuclear pore complex (NPC). RanT24N, in contrast to wild-type Ran-GDP, interacts only weakly with importin alpha and nucleoporins, and not at all with the import factor p10, consistent with its poor activity in nuclear import. However, RanT24N does interact stably with importin beta, Ran binding protein 1 and RCC1, an exchange factor for Ran. We show that Ran-GDP is essential for proper nuclear assembly and DNA replication, the requirement being primarily before the initiation of DNA replication. Ran-GDP therefore mediates the active transport of necessary factors or otherwise controls the onset of S-phase in this system. PMID- 9739076 TI - Centrosome dynamics in early embryos of Caenorhabditis elegans. AB - The early Caenorhabditis elegans embryo divides with a stereotyped pattern of cleavages to produce cells that vary in developmental potential. Differences in cleavage plane orientation arise between the anterior and posterior cells of the 2-cell embryo as a result of asymmetries in centrosome positioning. Mechanisms that position centrosomes are thought to involve interactions between microtubules and the cortex, however, these mechanisms remain poorly defined. Interestingly, in the early embryo the shape of the centrosome predicts its subsequent movement. We have used rhodamine-tubulin and live imaging techniques to study the development of asymmetries in centrosome morphology and positioning. In contrast to studies using fixed embryos, our images provide a detailed characterization of the dynamics of centrosome flattening. In addition, our observations of centrosome behavior in vivo challenge previous assumptions regarding centrosome separation by illustrating that centrosome flattening and daughter centrosome separation are distinct processes, and by revealing that nascent daughter centrosomes may become separated from the nucleus. Finally, we provide evidence that the midbody specifies a region of the cortex that directs rotational alignment of the centrosome-nucleus complex and that the process is likely to involve multiple interactions between microtubules and the cortex; the process of alignment involves oscillations and overshoots, suggesting a multiplicity of cortical sites that interact with microtubules. PMID- 9739077 TI - Developmental association of the beta-galactoside-binding protein galectin-1 with the nuclear matrix of rat calvarial osteoblasts. AB - The protein composition of the nuclear matrix changes significantly as the osteoblast matures from a proliferating pre-osteoblast to an osteocyte embedded in a mineralized matrix. These matrix protein are the result of developmental stage-specific gene expression during osteoblast differentiation. To isolate nuclear matrix proteins unique to the bone phenotype we analyzed nuclear matrix preparations from cultures of rat calvarial osteoblasts by high resolution two dimensional gel electrophoresis at two different stages: proliferation (day 3) and differentiation (day 18, mineralized). We characterized one protein (14 kDa; pI 5.0), that was detectable only in the nuclear matrix of differentiated osteoblasts. By mass spectrometry and microsequencing, this protein was identified as the beta -galactoside-binding protein galectin-1. Both immunofluorescence staining of nuclear matrix preparations with the galectin-1 antibody and western blot analysis of subcellular fractions confirmed that galectin-1 is only associated with the nuclear matrix in differentiated osteoblasts as the result of differential retention. Galectin-1 protein and mRNA are present throughout osteoblast differentiation. Galectin-1 is present in the cytoplasmic and nuclear fractions in both proliferating and differentiated osteoblasts. However, its only stable binding is to the nuclear matrix of the differentiated osteoblast; but, in proliferating osteoblasts, galectin-1 is not retained in the nuclear matrix. Taken together, our results suggest that developmental association of galectin-1 with the nuclear matrix reflects differential subnuclear binding of galectin-1 during osteoblast differentiation. PMID- 9739078 TI - VE-cadherin and desmoplakin are assembled into dermal microvascular endothelial intercellular junctions: a pivotal role for plakoglobin in the recruitment of desmoplakin to intercellular junctions. AB - Vascular endothelial cells assemble adhesive intercellular junctions comprising a unique cadherin, VE-cadherin, which is coupled to the actin cytoskeleton through cytoplasmic interactions with plakoglobin, beta-catenin and alpha -catenin. However, the potential linkage between VE-cadherin and the vimentin intermediate filament cytoskeleton is not well characterized. Recent evidence indicates that lymphatic and vascular endothelial cells express desmoplakin, a cytoplasmic desmosomal protein that attaches intermediate filaments to the plasma membrane in epithelial cells. In the present study, desmoplakin was localized to intercellular junctions in human dermal microvascular endothelial cells. To determine if VE-cadherin could associate with desmoplakin, VE-cadherin, plakoglobin, and a desmoplakin amino-terminal polypeptide (DP-NTP) were co expressed in L-cell fibroblasts. In the presence of VE-cadherin, both plakoglobin and DP-NTP were recruited to cell-cell borders. Interestingly, beta-catenin could not substitute for plakoglobin in the recruitment of DP-NTP to cell borders, and DP-NTP bound to plakoglobin but not beta-catenin in the yeast two-hybrid system. In addition, DP-NTP colocalized at cell-cell borders with alpha-catenin in the L cell lines, and endogenous desmoplakin and alpha-catenin colocalized in cultured dermal microvascular endothelial cells. This is in striking contrast to epithelial cells, where desmoplakin and alpha -+catenin are restricted to desmosomes and adherens junctions, respectively. These results suggest that endothelial cells assemble unique junctional complexes that couple VE-cadherin to both the actin and intermediate filament cytoskeleton. PMID- 9739079 TI - The IQGAP-related protein DGAP1 interacts with Rac and is involved in the modulation of the F-actin cytoskeleton and control of cell motility. AB - DGAP1 of Dictyostelium discoideum is a cell cortex associated 95 kDa protein that shows homology to both RasGTPase-activating proteins (RasGAPs) and RasGAP-related proteins. When tested for RasGAP activity, recombinant DGAP1 protein did not promote the GTPase activity of human H-Ras or of Dictyostelium RasG in vitro. Instead, DGAP1 bound to Dictyostelium Rac1A and human Rac1, but not to human Cdc42. DGAP1 preferentially interacted with the activated GTP-bound forms of Rac1 and Rac1A, but did not affect the GTPase activities. Since Rho-type GTPases are implicated in the formation of specific F-actin structures and in the control of cell morphology, the microfilament system of mutants that either lack or overexpress DGAP1 has been analysed. DGAP1-null mutants showed elevated levels of F-actin that was organised in large leading edges, membrane ruffles or numerous large filopods. Expression of actin fused to green fluorescent protein (GFP) was used to monitor the actin dynamics in these cells, and revealed that the F-actin cytoskeleton of DGAP1-null cells was rapidly re-arranged to form ruffles and filopods. Conversely, in DGAP1-overexpressing cells, the formation of cellular projections containing F-actin was largely suppressed. Measurement of cell migration demonstrated that DGAP1 expression is inversely correlated with the speed of cell motility. PMID- 9739080 TI - Regulation of mRNA localization by transmembrane signalling: local interaction of HB-GAM (heparin-binding growth-associated molecule) with the cell surface localizes beta-actin mRNA. AB - Localization of mRNAs is currently thought to be partially responsible for molecular sorting to specific compartments within the cell. In mammalian cells the best-studied example is the beta-actin mRNA that is localized to the cell processes, and its localization is necessary in migratory responses of cells. It is reasonable to assume that mRNA localization within cells is coupled to transmembrane signalling due to extracellular factors, but little is known about such putative mechanisms. We show here that HB-GAM, an extracellular matrix associated factor that enhances migratory responses in cells, is able to localize beta-actin mRNA when locally applied to cells via microbeads. The HB-GAM-induced mRNA localization is specifically inhibited by low concentrations of heparin and by heparitinase treatment of cells, showing that cell-surface heparin-type glycans are required for the effect. The finding that soluble N-syndecan is also inhibitory suggests that the transmembrane proteoglycan N-syndecan, previously identified as an HB-GAM receptor, is involved in the mRNA-localizing effect of HB GAM. Inhibition of the mRNA localization by the src-kinase inhibitor PP1 is compatible with an N-syndecan-mediated effect since the receptor function of N syndecan has been recently found to depend on the src-kinase signalling pathway. The mRNA-localizing activity of N-syndecan is also suggested by the finding that affinity-purified anti-N-syndecan antibodies coated on microbeads are able to localize beta-actin mRNA. PMID- 9739081 TI - Uptake by COPI-coated vesicles of both anterograde and retrograde cargo is inhibited by GTPgammaS in vitro. AB - On the basis of the cell surface protein CD8 we have constructed reporter molecules for both anterograde and retrograde transport from the Golgi complex. The cytoplasmic tail of CD8 was exchanged by a construct comprising a hemagglutinin (HA) epitope, the C-terminal sequence of the viral protein E19 (containing a KKXX retrieval signal) followed by a myc epitope (CD8-LT). Due to this masking of the KKXX retrieval signal CD8-LT is transported to the cell surface. Since the KKXX motif is joined to the myc epitope via a thrombin cleavage site, CD8-LT in isolated Golgi membranes can be proteolytically converted into an unmasked reporter molecule for retrograde transport (CD8-ST) in vitro. A CHO cell line stably expressing CD8-LT was generated and used for the isolation of Golgi membranes. These membranes were shown to contain CD8-LT en route to the cell surface. By addition of thrombin, CD8-LT could be efficiently converted into CD8-ST, and this allows us to study the sorting into coat protein COPI-coated vesicles of these different kinds of cargo on a comparative basis. COPI-coated vesicles were generated in vitro from Golgi membranes containing either CD8-LT or CD8-ST. When the incubation was performed in the presence of GTP, both CD8-LT and CD8-ST were packaged into COPI-coated vesicles. However, COPI-coated vesicles generated in the presence of the slowly hydrolyzable analogue of GTP, GTP(&ggr ;)S contained strikingly lower amounts of CD8-LT and CD8-ST. While COPI-coated vesicles accumulated about 12-fold in the presence of GTPgammaS these vesicles together contained only one fifth of cargo compared to the few vesicles generated in the absence of GTPgammaS. These data indicate that cargo packaging into COPI-coated vesicles requires hydrolysis of GTP. PMID- 9739082 TI - A cell cycle regulated MAP kinase with a possible role in cytokinesis in tobacco cells. AB - Mitogen-activated protein (MAP) kinases have been demonstrated to have a role in meiosis but their involvement in mitotic events is less clear. Using a peptide antibody raised against the tobacco MAP kinase p43(Ntf6) and extracts from synchronized tobacco cell suspension cultures, we show that this kinase is activated specifically during mitosis. Entry into mitosis appears to be necessary for the activation of the kinase, which occurs as a post-translational event. The activation of the kinase occurs in late anaphase/early telophase. The p43(Ntf6) protein shows a transient localization to the cell plate in anaphase cells, in the middle of the two microtubule arrays characteristic of the phragmoplast, a plant-specific structure involved in laying down the new cell wall. The combined data support a role for the MAP kinase p43(Ntf6) in cytokinesis in tobacco cells. PMID- 9739083 TI - Cdc18p can block mitosis by two independent mechanisms. AB - The DNA replication checkpoint is required to maintain the integrity of the genome, inhibiting mitosis until S phase has been successfully completed. The checkpoint preventing premature mitosis in Schizosaccharomyces pombe relies on phosphorylation of the tyrosine-15 residue on cdc2p to prevent its activation and hence mitosis. The cdc18 gene is essential for both generating the DNA replication checkpoint and the initiation of S phase, thus providing a key role for the overall control and coordination of the cell cycle. We show that the C terminus of the protein is capable of both initiating DNA replication and the checkpoint function of cdc18p. The C terminus of cdc18p acts upstream of the DNA replication checkpoint genes rad1, rad3, rad9, rad17, hus1 and cut5 and requires the wee1p/mik1p tyrosine kinases to block mitosis. The N terminus of cdc18p can also block mitosis but does so in the absence of the DNA replication checkpoint genes and the wee1p/mik1p kinases therefore acting downstream of these genes. Because the N terminus of cdc18p associates with cdc2p in vivo, we suggest that by binding the cdc2p/cdc13p mitotic kinase directly, it exerts an effect independently of the normal checkpoint control, probably in an unphysiological manner. PMID- 9739084 TI - Yel013p (Vac8p), an armadillo repeat protein related to plakoglobin and importin alpha is associated with the yeast vacuole membrane. AB - Proteins of the armadillo family are involved in diverse cellular processes in higher eukaryotes. Some of them, like armadillo, beta-catenin and plakoglobins have dual functions in intercellular junctions and signalling cascades. Others, belonging to the importin-alpha-subfamily are involved in NLS recognition and nuclear transport, while some members of the armadillo family have as yet unknown functions. Here, we introduce the Saccharomyces cerevisiae protein Yel013p as a novel armadillo (arm) repeat protein. The ORF Yel013w was identified in the genome project on chromosome V (EMBL: U18530) and codes for an acidic protein of 578 residues with 8 central arm-repeats, which are closely related to the central repeat-domain of Xenopus laevis plakoglobin. We show that Yel013p (Vac8p) is constitutively expressed in diploid and haploid yeasts and that it is not essential for viability and growth. However, the vacuoles of mutant cells are multilobular or even fragmented into small vesicles and the processing of aminopeptidase I, representing the cytoplasm-to-vacuole transport pathway, is strongly impaired. Consistent with these observations, subcellular fractionation experiments, immunolocalization and expression of green fluorescent protein (GFP) fusion proteins revealed that Yel013p (Vac8p) is associated with the vacuolar membrane. Our data provide evidence for the involvement of an arm-family member in vacuolar morphology and protein targeting to the vacuole. PMID- 9739085 TI - Integrin ligation and PKC activation are required for migration of colon carcinoma cells. AB - The activation of protein kinases C (PKCs) is an essential step in integrin dependent cell adhesion and spreading. In this report we examined the effect of the phorbol ester PMA, a PKC activator, on adhesion, spreading and migration of a colon carcinoma cell line, HT29-D4. Treatment with PMA increased the rate of cell spreading and induced the migration of these cells towards purified matrix proteins in haptotaxis assays on Boyden chambers. PMA-induced effects were the result of PKCs activation, as shown by using the inactive isomer 4alpha-PMA and PKCs inhibitors. The involvement of integrins in the phorbol ester-induced cell migration was demonstrated both by the absence of migration of cells plated on membranes coated with poly-L-lysine and by the use of function blocking antibodies. Thus, interactions between alpha 2beta1, alpha3beta1, alpha6beta4, alpha vbeta5, alphavbeta6 integrins and their specific ligands are necessary for the PKC-mediated migration. However, adhesion, immunoprecipitation and immunocytofluorometry experiments clearly showed that HT29-D4 cell haptotaxis induced by PKC activation is not a consequence of quantitative or qualitative changes in the cell surface integrins. We also demonstrated that PKCs were able to activate the MAP kinase pathway and that the impediment of MAP kinase activation resulted in the loss of cell migration. Moreover, stimulation of the insulin-like growth factor I signalling pathway led to MAP kinase activation and to the induction of cell migration. In addition, the growth factor-induced motility of HT29-D4 cells was affected both by PKC and MAP kinase cascade inhibitors. It thus appears that both integrin ligation and MAP kinase activation by PKCs are required to promote the migration of HT29-D4 cells. PMID- 9739086 TI - A domain flip as a result of a single amino-acid substitution. AB - BACKGROUND: The self-assembly properties of beta domains are important features of diverse classes of proteins that include cell-adhesion molecules, surface receptors and the immunoglobulin superfamily. Immunoglobulin light-chain variable domains are well suited to the study of structural factors that determine dimerization, including how residues at the interface influence the preferred dimer arrangement. RESULTS: Single-site mutants of light-chain variable domain Len, designated LenQ38E and LenK30T, formed 'flipped' dimers in which one domain was rotated by about 180 degrees compared with the native protein. The dimer in the native protein is similar to that found between variable domains in Fab immunoglobulin fragments. When compared to the native dimer, more surface area is buried, and more hydrogen bonds and salt bridges are formed between the monomers in the flipped conformation. CONCLUSIONS: Immunoglobulin light-chain variable domains can form a minimum of two distinct quaternary structures. Single-site mutations resulting from changes of one base, such as the exchange of Gln38 to Glu or Lys30 to Thr, change the 'conventional' dimer of protein Len to a flipped arrangement. Native Len is not found in the flipped-domain dimer conformation because it would have excess positive electrostatic potential at the dimer interface that is not compensated by other forces. Excess negative or positive electrostatic potential at the dimer interface can have a determining effect on the mode of dimerization. PMID- 9739087 TI - The crystal structure of dienoyl-CoA isomerase at 1.5 A resolution reveals the importance of aspartate and glutamate sidechains for catalysis. AB - BACKGROUND: The degradation of unsaturated fatty acids is vital to all living organisms. Certain unsaturated fatty acids must be catabolized via a pathway auxiliary to the main beta-oxidation pathway. Dienoyl-coenzyme A (dienoyl-CoA) isomerase catalyzes one step of this auxiliary pathway, the isomerization of 3 trans,5-cis-dienoyl-CoA to 2-trans,4-trans-dienoyl-CoA, and is imported into both mitochondria and peroxisomes. Dienoyl-CoA isomerase belongs to a family of CoA binding proteins that share the enoyl-CoA hydratase/isomerase sequence motif. RESULTS: The crystal structure of rat dienoyl-CoA isomerase has been determined at 1.5 A resolution. The fold closely resembles that of enoyl-CoA hydratase and 4 chlorobenzoyl-CoA dehalogenase. Dienoyl-CoA isomerase forms hexamers made up of two trimers. The structure contains a well ordered peroxisomal targeting signal type-1 which is mostly buried in the inter-trimer space. The active-site pocket is deeply buried and entirely hydrophobic, with the exception of the acidic residues Asp176, Glu196 and Asp204. Site-directed mutagenesis of Asp204 revealed that this residue is essential for catalysis. In a molecular modeling simulation, a molecule of 3-trans,5-cis-octadienoyl-CoA was docked into the active site. CONCLUSIONS: The structural data, supported by the mutagenesis data, suggest a reaction mechanism where Glu196 acts as a proton acceptor and Asp204 acts as a proton donor. Asp176 is paired with Glu196 and is important for optimizing the catalytic proton transfer properties of Glu196. In the predicted mode of substrate binding, an oxyanion hole stabilizes the transition state by binding the thioester oxygen. The presence of a buried peroxisomal targeting signal suggests that dienoyl-CoA isomerase is prevented from reaching its hexameric structure in the cytosol. PMID- 9739088 TI - Structure of 3-isopropylmalate dehydrogenase in complex with 3-isopropylmalate at 2.0 A resolution: the role of Glu88 in the unique substrate-recognition mechanism. AB - BACKGROUND: 3-Isopropylmalate dehydrogenase (IPMDH) and isocitrate dehydrogenase (ICDH) belong to a unique family of bifunctional decarboxylating dehydrogenases. Although the ICDH dimer catalyzes its reaction under a closed conformation, known structures of the IPMDH dimer (without substrate) adopt a fully open or a partially closed form. Considering the similarity in the catalytic mechanism, the IPMDH dimer must be in a fully closed conformation during the reaction. A large conformational change should therefore occur upon substrate binding. RESULTS: We have determined the crystal structure of IPMDH from Thiobacillus ferrooxidans (Tf) complexed with 3-isopropylmalate (IPM) at 2.0 A resolution by the molecular replacement method. The structure shows a fully closed conformation and the substrate-binding site is quite similar to that of ICDH except for a region around the gamma-isopropyl group. The gamma group is recognized by a unique hydrophobic pocket, which includes Glu88, Leu91 and Leu92 from subunit 1 and Val193' from subunit 2. CONCLUSIONS: A large movement of domain 1 is induced by substrate binding, which results in the formation of the hydrophobic pocket for the gamma-isopropyl moiety of IPM. A glutamic acid in domain 1, Glu88, participates in the formation of the hydrophobic pocket. The C beta and C gamma atoms of Glu88 interact with the gamma-isopropyl moiety of IPM and are central to the recognition of substrate. The acidic tip of Glu88 is likely to interact with the nicotinamide mononucleotide (NMN) ribose of NAD+ in the ternary complex. This structure clearly explains the substrate specificity of IPMDH. PMID- 9739089 TI - Crystal structure of JNK3: a kinase implicated in neuronal apoptosis. AB - BACKGROUND: The c-Jun N-terminal kinases (JNKs) are members of the mitogen activated protein (MAP) kinase family, and regulate signal transduction in response to environmental stress. Activation and nuclear localization of JNK3, a neuronal-specific isoform of JNK, has been associated with hypoxic and ischemic damage of CA1 neurons in the hippocampus. Knockout mice lacking JNK3 showed reduced apoptosis of hippocampal neurons and reduced seizure induced by kainic acid, a glutamate-receptor agonist. Thus, JNK3 may be important in the pathology of neurological disorders and is of significant medical interest. RESULTS: We report here the structure of unphosphorylated JNK3 in complex with adenylyl imidodiphosphate, an ATP analog. JNK3 has a typical kinase fold, with the ATP binding site situated within a cleft between the N- and C-terminal domains. In contrast to other known MAP kinase structures, the ATP-binding site of JNK3 is well ordered; the glycine-rich nucleotide-binding sequence forms a beta-strand turn-beta-strand structure over the nucleotide. Unphosphorylated JNK3 assumes an open conformation, in which the N- and C-terminal domains are twisted apart relative to their positions in cAMP-dependent protein kinase. The rotation leads to the misalignment of some of the catalytic residues. The phosphorylation lip of JNK3 partially blocks the substrate-binding site. CONCLUSIONS: This is the first JNK structure to be determined, providing a unique opportunity to compare structures from the three MAP kinase subfamilies. The structure reveals atomic level details of the shape of JNK3 and the interactions between the kinase and the nucleotide. The misalignment of catalytic residues and occlusion of the active site by the phosphorylation lip may account for the low activity of unphosphorylated JNK3. The structure provides a framework for understanding the substrate specificity of different JNK isoforms, and should aid the design of selective JNK3 inhibitors. PMID- 9739090 TI - The solution structure of an RNA loop-loop complex: the ColE1 inverted loop sequence. AB - BACKGROUND: Replication of the ColE1 plasmid of Escherichia coli is regulated by the interaction of sense and antisense plasmid-encoded transcripts. The antisense RNA I negatively regulates the replication of the plasmid by duplex formation with complementary RNA II. The interaction is initiated by the formation of a double helix between seven-nucleotide loops from each RNA and is stabilized by binding of the RNA one modulator (ROM) protein. The ROM protein is thought to recognize a specific RNA structure, regardless of sequence. RESULTS: The solution structure of a loop-loop complex between model RNA hairpins that resemble RNA I and RNA II has been determined by nuclear magnetic resonance spectroscopy. The model hairpins have loop sequences inverted 5' to 3' relative to the wild-type sequence and were chosen because of their complex's slow dissociation in comparison to the wild type. The complex has continuous stacking from the 3'-side of one stem helix through the loop-loop helix to the other stem helix. One residue from each hairpin has a unique phosphodiester bond which bridges and narrows the major groove. These bridging phosphates are in close proximity to the phosphate groups of the adjacent bases, forming unique structural motifs called phosphate clusters. The purine residue at the 3'-end of the loop-loop helix of one RNA stacks on a purine residue on the 5'-side of the other RNA stem, and there are strong cross-strand stacking interactions between guanine bases in the stem helices adjacent to the loops. CONCLUSIONS: Unique distortions, such as the strong bend and the phosphate clusters flanking the major groove of the loop-loop helix, provide an attractive nonsequence-specific structural feature for recognition by the ROM protein. The structure provides a basis for rationalizing the sequence dependence of the stability of loop-loop interaction. PMID- 9739091 TI - Phosducin induces a structural change in transducin beta gamma. AB - BACKGROUND: Phosducin binds tightly to the beta gamma subunits (Gt beta gamma) of the heterotrimeric G protein transducin, preventing Gt beta gamma reassociation with Gt alpha-GDP and thereby inhibiting the G-protein cycle. Phosducin-like proteins appear to be widely distributed and may play important roles in regulating many heterotrimeric G-protein signaling pathways. RESULTS: The 2.8 A crystal structure of a complex of bovine retinal phosducin with Gt beta gamma shows how the two domains of phosducin cover one side and the top of the seven bladed beta propeller of Gt beta gamma. The binding of phosducin induces a distinct structural change in the beta propeller of Gt beta gamma, such that a small cavity opens up between blades 6 and 7. Electron density in this cavity has been assigned to the farnesyl moiety of the gamma subunit. CONCLUSIONS: beta gamma subunits of heterotrimeric G proteins can exist in two distinct conformations. In the R (relaxed) state, corresponding to the structure of the free beta gamma or the structure of beta gamma in the alpha beta gamma heterotrimer, the hydrophobic farnesyl moiety of the gamma subunit is exposed, thereby mediating membrane association. In the T (tense) state, as observed in the phosducin-Gt beta gamma structure, the farnesyl moiety of the gamma subunit is effectively buried in the cavity formed between blades 6 and 7 of the beta subunit. Binding of phosducin to Gt beta gamma induces the formation of this cavity, resulting in a switch from the R to the T conformation. This sequesters beta gamma from the membrane to the cytosol and turns off the signal-transduction cascade. Regulation of this membrane association/dissociation switch of Gt beta gamma by phosducin may be a general mechanism for attenuation of G protein coupled signal transduction cascades. PMID- 9739092 TI - How a protein prepares for B12 binding: structure and dynamics of the B12-binding subunit of glutamate mutase from Clostridium tetanomorphum. AB - BACKGROUND: Glutamate mutase is an adenosylcobamide (coenzyme B12) dependent enzyme that catalyzes the reversible rearrangement of (2S)-glutamate to (2S,3S)-3 methylaspartate. The enzyme from Clostridium tetanomorphum comprises two subunits (of 53.7 and 14.8 kDa) and in its active form appears to be an alpha 2 beta 2 tetramer. The smaller subunit, termed MutS, has been characterized as the B12 binding component. Knowledge on the structure of a B12-binding apoenzyme does not exist. RESULTS: The solution structure and important dynamical aspects of MutS have been determined from a heteronuclear NMR study. The global fold of MutS in solution resembles that determined by X-ray crystallography for the B12-binding domains of Escherichia coli methionine synthase and Propionibacterium shermanii methylmalonyl CoA mutase. In these two proteins a histidine residue displaces the endogenous cobalt-coordinating ligand of the B12 cofactor. In MutS, however, the segment of the protein containing the conserved histidine residue forms part of an unstructured and mobile extended loop. CONCLUSIONS: A comparison of the crystal structures of two B12-binding domains, with bound B12 cofactor, and the solution structure of the apoprotein MutS has helped to clarify the mechanism of B12 binding. The major part of MutS is preorganized for B12 binding, but the B12 binding site itself is only partially formed. Upon binding B12, important elements of the binding site appear to become structured, including an alpha helix that forms one side of the cleft accommodating the nucleotide 'tail' of the cofactor. PMID- 9739093 TI - Adding 'splice' to protein engineering. PMID- 9739094 TI - The crystal structure of the Leishmania major surface proteinase leishmanolysin (gp63). AB - BACKGROUND: Despite their medical importance, there is little available structural information for the surface antigens of infectious protozoa. Diseases caused by the protozoan parasite Leishmania are common in many developing countries. Human infection occurs during the bite of infected sandfilies, when Leishmania promastigote cells from the insect gut enter the bloodstream. Promastigotes in the blood parasitize macrophages, often causing serious disease. Leishmanolysin is the predominant protein surface antigen of promastigotes, and is assumed to have a key role during infection. Leishmanolysin is a membrane bound zinc proteinase, active in situ. Similar molecules exist in other trypanomastid protozoa. RESULTS: Two crystal forms of leishmanolysin were obtained from protein purified from promastigote membranes. A single lead derivative in both crystal forms was used to solve the structure. The structure reveals three domains, two of which have novel folds. The N-terminal domain has a similar structure to the catalytic modules of zinc proteinases. The structure clearly shows that leishmanolysin is a member of the metzincin class of zinc proteinases. CONCLUSIONS: The unexpected metzincin features of the leishmanolysin structure suggest that the metzincin fold may be more widespread than indicated by sequence homologies amongst existing metzincin zinc proteinases. The similarity of the active-site structure to previously well characterized metzincin class zinc proteinases should aid the development of specific inhibitors. These inhibitors might be used to determine the function of leishmanolysin in the insect and during mammalian infection, and may aid the development of drugs for human leishmaniasis. PMID- 9739095 TI - Involvement of the C terminus in intramolecular nitrogen channeling in glucosamine 6-phosphate synthase: evidence from a 1.6 A crystal structure of the isomerase domain. AB - BACKGROUND: Glucosamine 6-phosphate synthase (GlmS) catalyses the first step in hexosamine metabolism, converting fructose-6P (6 phosphate) into glucosamine-6P using glutamine as a nitrogen source. GlmS is a bienzyme complex consisting of two domains that catalyse glutamine hydrolysis and sugar-phosphate isomerisation, respectively. Knowledge of the three-dimensional structure of GlmS is essential for understanding the general principles of catalysis by ketol isomerases and the mechanism of nitrogen transfer in glutamine amidotransferases. RESULTS: The crystal structure of the isomerase domain of the Escherichia coli GlmS with the reaction product, glucosamine-6P, has been determined at 1.57 A resolution. It is comprised of two topologically identical subdomains, each of which is dominated by a nucleotide-binding motif of a flavodoxin type. The catalytic site is assembled by dimerisation of the protein. CONCLUSIONS: The isomerase active site of GlmS seems to be the result of evolution through gene duplication and subsequent dimerisation. Isomerisation of fructose-6P is likely to involve the formation of a Schiff base with Lys603 of the enzyme, the ring-opening step catalysed by His504, and the proton transfer from C1 to C2 of the substrate effected by Glu488. The highly conserved C-terminal fragment of the chain may play a key role in substrate binding, catalysis and communication with the glutaminase domain. The corresponding sequence pattern DXPXXLAK[SC]VT (in single letter amino-acid code, where X is any amino acid and letters in brackets indicate that either serine or cysteine may take this position) may be considered as a fingerprint of GlmS. PMID- 9739096 TI - HIV gp120: double lock strategy foils host defences. AB - The recent determination of the structure of a complex formed between the HIV-1 glycoprotein gp120, CD4 and an antibody fragment has revealed new mechanisms for viral evasion of the immune response and shed light on how the virus enters target cells. The results of this work, together with related biochemical studies, may assist in the future design of therapeutic strategies against HIV-1 infection. PMID- 9739097 TI - Solution structure of the DNA-binding domain of human telomeric protein, hTRF1. AB - BACKGROUND: Mammalian telomeres consist of long tandem arrays of the double stranded TTAGGG sequence motif packaged by a telomere repeat binding factor, TRF1. The DNA-binding domain of TRF1 shows sequence homology to each of three tandem repeats of the DNA-binding domain of the transcriptional activator c-Myb. The isolated c-Myb-like domain of human TRF1 (hTRF1) binds specifically to telomeric DNA as a monomer, in a similar manner to that of homeodomains. So far, the only three-dimensional structure of a telomeric protein to be determined is that of a yeast telomeric protein, Rap 1p. The DNA-binding domain of Rap 1p contains two subdomains that are structurally closely related to c-Myb repeats. We set out to determine the solution structure of the DNA-binding domain of hTRF1 in order to establish its mode of DNA binding. RESULTS: The solution structure of the DNA-binding domain of hTRF1 has been determined and shown to comprise three helices. The architecture of the three helices is very similar to that of each Rap 1p subdomain and also to that of each c-Myb repeat. The second and third helix form a helix-turn-helix (HTH) variant. The length of the third helix of hTRF1 is similar to that of the second subdomain of Rap 1p. CONCLUSIONS: The hTRF1 DNA-binding domain is likely to bind to DNA in a similar manner to that of the second subdomain of Rap 1p. On the basis of the Rap 1p-DNA complex, a model of the hTRF1 DNA-binding domain in complex with human telomeric DNA was constructed. In addition to DNA recognition by the HTH variant, a flexible N terminal arm of hTRF1 is likely to interact with DNA. PMID- 9739098 TI - A single dose model of methamphetamine-induced neurotoxicity in rats: effects on neostriatal monoamines and glial fibrillary acidic protein. AB - The neurotoxic effects of a single administration of methamphetamine (MA) were studied under conditions conducive to MA-induced hyperthermia. After a single dose of MA (10, 20, 30, or 40 mg/kg, s. c.) or saline (3 ml/kg) to Sprague-Dawley CD rats, rectal temperatures were monitored for 9 h in a room with an ambient temperature of 22.0+/-0.5 degrees C. MA induced significant dose-dependent hyperthermia, however, no significant increase in mortality occurred. Neostriatal DA, 5-HT, TH, and GFAP were assayed 3 days following treatment. MA induced dose dependent reductions of DA, 5-HT and TH, and increased GFAP. For DA, at doses of 20, 30, or 40 mg/kg the reductions were to 71%, 49%, and 29%, and for 5-HT were to 73%, 44%, and 19% of control values. No reductions were seen after the 10 mg/kg dose. Semiquantitative analysis Western blots of TH and GFAP demonstrated that TH was reduced to 52%, 75%, and 28%, and GFAP was increased to 125%, 134%, and 149% of control values at MA doses of 20, 30, or 40 mg/kg, respectively. No significant changes in TH or GFAP were seen at the 10 mg/kg MA dose. These results demonstrate that a single-dose of MA can be as effective as the widely used four-dose every 2 h regimen. Moreover, mortality can be minimized by monitoring core body temperature and preventing MA-induced hyperthermia from exceeding 41.5 degrees C. PMID- 9739099 TI - Signalling interactions during facial development. AB - The development of the vertebrate face is a dynamic multi-step process which starts with the formation of neural crest cells in the developing brain and their subsequent migration to form, together with mesodermal cells, the facial primordia. Signalling interactions co-ordinate the outgrowth of the facial primordia from buds of undifferentiated mesenchyme into the intricate series of bones and cartilage structures that, together with muscle and other tissues, form the adult face. Some of the molecules that are thought to be involved have been identified through the use of mouse mutants, data from human craniofacial syndromes and by expression studies of signalling molecules during facial development. However, the way that these molecules control the epithelial mesenchymal interactions which mediate facial outgrowth and morphogenesis is unclear. The role of neural crest cells in these processes has also not yet been well defined. In this review we discuss the complex interaction of all these processes during face development and describe the candidate signalling molecules and their possible target genes. PMID- 9739100 TI - In situ mRNA hybridization study of the distribution of choline acetyltransferase in the human brain. AB - We examined the distribution of choline acetyltransferase (ChAT) mRNA in the brain of six autopsied individuals by in situ hybridization with 35S-labeled human ChAT riboprobes. Neurons containing hybridization signal for ChAT mRNA were observed in the nucleus of the diagonal band of Broca, the basal nucleus of Meynert, the caudate nucleus, the putamen, the pedunculopontine tegmental nucleus, the laterodorsal tegmental nucleus, the parabigeminal nucleus, the oculomotor nucleus and the trochlear nucleus. These findings were in good agreement with previous ChAT-immunohistochemical data. In contrast, labeled neurons were not observed in the medial septal and medial habenular nuclei, in which previously ChAT-immunoreactive neurons have been identified in many mammalian species, including the human. An unexpected result of the present study was the demonstration of neurons with ChAT mRNA signal in restricted areas of the human cerebral cortex. PMID- 9739101 TI - Electron microscopic analysis of gamma-aminobutyric acid and glycine colocalization in rat trigeminal subnucleus caudalis. AB - Postembedding immunogold methods were used to examine the distribution of gamma aminobutyric acid (GABA) and glycine and especially their colocalization in glomerular neuronal profiles adjacent to trigeminal primary afferent profiles in lamina II of rat subnucleus caudalis. We found that 60% of the profiles adjacent to the trigeminal primary afferent terminals exhibited colocalization of GABA and glycine. GABA alone was found to localize in 17% of the adjacent profiles. Glycine alone was found to localize in 18% of the adjacent profiles. Of interest, 10% of the trigeminal primary afferent fibers showed glycine localization. All the profiles with colocalization of GABA and glycine were identified as presynaptic axonal terminals, suggesting a possible cumulative effect by these two inhibitory neurotransmitters in presynaptic inhibition. These findings show that GABA and glycine colocalize in a subpopulation of presynaptic axonal terminals within lamina II of the subnucleus caudalis. The possible origins of these axons are discussed, as well as their potential involvement in presynaptic inhibition of orofacial nociception. PMID- 9739102 TI - Independence of, and interactions between, cannabinoid and opioid signal transduction pathways in N18TG2 cells. AB - N18TG2 neuroblastoma cells co-express delta-opioid and CB1-cannabinoid receptors. Both receptors are negatively coupled to adenylyl cyclase through pertussis toxin sensitive GTP-binding proteins. In the present study, we confirmed the independent activity of opioid and cannabinoid agonists, and investigated chronic interactions between the two signal transduction pathways in these cells. Opioid and cannabinoid agonists stimulated [35S]guanosine-5'-O-(3-thiotriphosphate) binding to N18TG2 membranes. When the opioid agonist etorphine and the cannabinoid agonist desacetyllevonantradol (DALN) were applied together, the stimulation was similar to the arithmetic sum of the two separate effects. This additivity existed even after partial ablation of the G-proteins reservoir with a low concentration of pertussis toxin, indicating that opioid and cannabinoid receptors activate different pools of G-proteins in N18TG2 cells. Chronic treatment of the cells with either opioid or cannabinoid agonists induced desensitization to the respective drug. In addition, asymmetric cross desensitization was found: while long-term exposure to DALN induced homologous desensitization, and did not reduce the effect of etorphine, long-term exposure to etorphine attenuated the cannabinoid activation of G-proteins. Chronic exposure to either DALN or etorphine not only induced desensitization, but also elevated the basal activity of G-proteins in the exposed cells. The combination of the two drugs did not yield an additive activation, suggesting that chronic exposure of N18TG2 cultures to cannabinoid and opioid agonists modified a common responding element within the cells. This work presents the N18TG2 neuroblastoma as a suitable experimental model to study the molecular mechanism(s) underlying chronic interactions between opioid and cannabinoid drugs. PMID- 9739103 TI - Developmental expression patterns of mouse sFRP genes encoding members of the secreted frizzled related protein family. AB - Development of the metanephric kidney is an experimental model system to analyze interactions between mesenchymal and epithelial cells and mesenchymal-epithelial transition. To study the underlying genetic mechanisms we employed organ culture and differential display PCR to identify genes regulated upon induction of mesenchymal cells. One of the genes found encodes the secreted frizzled related protein 2 (sFRP2) that is upregulated within 2 days of in vitro development. In vivo sFRP2 expression was likewise found in mesenchymal condensates and subsequent epithelial structures. Detailed in situ hybridization analysis revealed sFRP2 expression during development of the eye, brain, neural tube, craniofacial mesenchyme, joints, testis, pancreas and below the epithelia of oesophagus, aorta and ureter where smooth muscles develop. In a comparative analysis transcripts of the related sFRP1 and sFRP4 genes were frequently found in the same tissues as sFRP2 with their expression domains overlapping in some instances, but mutually exclusive in others. While sFRP1 is specifically expressed in the embryonic metanephros, eye, brain, teeth, salivary gland and small intestine, there is only weak expression of sFRP4 except for the developing teeth, eye and salivary gland. The interpretation of the highly specific spatial and temporal expression patterns of sFRP genes will partly depend on a better functional understanding of the interaction between wnt, fz and sFRP family members. Nevertheless, sFRP genes must play quite distinct roles in the morphogenesis of several organ systems. PMID- 9739104 TI - Norepinephrine inhibits neurons of the intermediate subnucleus of the lateral septum via alpha2-adrenoceptors. AB - The physiological and pharmacological actions of norepinephrine (NE) on neurons of the intermediate subnucleus of the lateral septum (LSI) were examined using intracellular recordings in rat brain-slices. Bath-applied NE inhibited 72.5%, excited 5.5% and had no effect on 22% of LSI neurons tested; this study focused on the inhibitory effects of NE. In current clamp recordings, 100 microM NE produced a hyperpolarization of 10.82+/-0.72 mV (n=84) with a decrease in input resistance. In voltage-clamp, NE produced a direct, post-synaptic outward current of 206.8+/-22 pA (n=37) with a 64. 3+/-4.9% increase in input conductance (IC50 17.7+/-4 microM). The NE-induced inhibition was mimicked by the alpha2-agonist, UK14,304, but not by the alpha1- or beta-adrenoceptor agonists. The alpha2 agonist, clonidine, had a weak effect in LSI neurons. Interestingly, the magnitude of the UK14,304-induced response varied between cells (ranging from 29.5 to 320% of the maximal NE inhibition), possibly suggesting the involvement of alpha2A-(high affinity for UK14,304) and non-alpha2A (low affinity for UK14,304) adrenoceptor subtypes. While the alpha2-antagonists, yohimbine, rauwolscine and idazoxan blocked NE-induced inhibition in all neurons tested, the prototypical alpha1-antagonist, prazosin produced a variable degree of block (9 58%), further indicating the possible involvement of alpha2A (prazosin insensitive) and non-alpha2A (prazosin-sensitive) receptors. However a lack of more selective pharmacological tools precludes definitive classification of the alpha2-receptor-mediated responses into different subtypes. The alpha2-receptor mediated current in LSI neurons displayed Ba2+-sensitive inward rectification, reversed polarity near EK and was sensitive to external K+. In conclusion, NE inhibits LSI neurons via alpha2-adrenoceptor subtypes. PMID- 9739105 TI - Xenopus Zic family and its role in neural and neural crest development. AB - We characterized two new members of the Zic family, Xenopus Zic1 and Zic2. They are very similar to mouse Zic1 and Zic2 in the protein coding region including the zinc finger domain. In early gastrula, Zic1 expression was restricted to the prospective neural plate region whereas Zic2 was expressed widely in the ectoderm. We observed enhanced neural and neural crest-derived tissue formation in the Zic1 or Zic2 overexpressed embryos and neural and neural crest marker induction in the Zic1 or Zic2 overexpressed animal cap explants. Our findings suggest that Zic1 and Zic2 have essentially the same properties as Zic3 and that the Xenopus Zic family may act cooperatively in the initial phase of neural and neural crest development. PMID- 9739106 TI - Genetic rescue of segmentation defect in MesP2-deficient mice by MesP1 gene replacement. AB - Gene knock-out and knock-in strategies are employed to investigate the function of MesP1. MesP1 belongs to the same family of bHLH transcription factors as MesP2. The early expression pattern observed in the early mesoderm at the onset of gastrulation is restricted to Mesp1, while the later expression pattern in the anterior presomitic mesoderm during somitogenesis is almost the same for Mesp1 as for Mesp2. Homozygous Mesp1 null mice exhibited growth retardation after 7.5 dpc and died before 10.5 dpc with many developmental defects. The function of MesP1 during somitogenesis was not clearly revealed because of their early death and the possible compensation by MesP2. In order to examine the functions of MesP1 during somitogenesis, we replaced the Mesp2 gene with Mesp1 cDNA, using a gene knock-in strategy. The introduced Mesp1 cDNA could rescue the defects caused by Mesp2 deficiency in a dosage-dependent manner. Mice which lacked Mesp2 expression but had four copies of the Mesp1 gene survived into the adulthood and were fertile. The skeletal defects and the reduction in expression of Notch1, Notch2 and FGFR-1 previously observed in Mesp2 null mice were almost completely rescued by the introduced MesP1. Thus, it is concluded that the functions of MesP1 during somitogenesis, like MesP2, are also mediated via notch-delta and FGF signaling systems. PMID- 9739107 TI - Brain vasopressin levels in Down syndrome and Alzheimer's disease. AB - Performing gene hunting in Down Syndrome fetal brain we detected an overexpressed sequence highly homologous to the human vasopressin gene. As this neuropeptide may be involved in the pathogenetic mechanism and, moreover, was described to play a role in memory and learning, we decided to study the brain gene product level in Down Syndrome (DS), controls and patients with Alzheimer's disease (AD). Subtractive hybridization was used to study the differential expression between steady state mRNA levels in fetal brain of DS and controls at the 23rd week of gestation. A radioimmunological method was used to determine vasopressin (AVP) in five brain regions of each 9 aged DS brains, 9 brains with AD and 9 control individuals, obtained from brain bank. An overexpressed nucleic acid sequence with 91% homology to the vasopressin gene was detected in both fetal brains with DS. AVP levels in controls were of the order cerebellum>occipital>frontal>parietal>temporal lobe and were significantly higher in temporal lobe and lower in cerebellum of patients with DS. AVP levels in brain of AD patients were also significantly increased in temporal lobe but were not reduced in cerebellum. The biological meaning of increased AVP remain unclear but may be linked to the neurodegenerative processes, proposed to be similar in both disorders. Data from gene hunting in fetal DS brain along with our data on aged DS and AD patients suggest the early involvement of AVP in the pathomechanism accompanying cholinergic, monoaminergic and neuropeptidergic deficits described in DS and AD. PMID- 9739108 TI - Strands of embryonic mesencephalic tissue show greater dopamine neuron survival and better behavioral improvement than cell suspensions after transplantation in parkinsonian rats. AB - The success of embryonic neural transplants as a treatment for patients with Parkinson's disease has been limited by poor survival of transplanted dopamine neurons. To see if a new partially intact tissue preparation method improves survival, we have developed a technique for extruding embryonic tissue into strands. We expected this method to reduce cell damage and improve transplant survival as well as provide improved tissue delivery. We have compared transplants of tissue strands with mechanically dispersed suspensions of embryonic day 15 rat ventral mesencephalon. Tissue from ventral mesencephalon was transplanted into a single site in dopamine denervated striatum of unilateral 6 hydroxydopamine (6-OHDA) lesioned rats. To evaluate the effects of striatal cografts and growth factors on dopamine cell survival, dispersed mesencephalic cells were cotransplanted with dispersed striatal cells. Another group had dispersed mesencephalic cells cotransplanted with striatal cells incubated in the cold for 2 h with glial cell line-derived neurotrophic factor (GDNF, 100 ng/ml), insulin-like growth factor-I (IGF-I, 1500 ng/ml), and basic fibroblast growth factor (bFGF, 150 ng/ml). Behavioral improvement was assessed monthly by changes in methamphetamine-induced rotational behavior. Animals were sacrificed after 3 months, and dopamine neurons were identified by tyrosine hydroxylase (TH) immunohistochemistry. Transplants of tissue strands produced better dopamine neuron survival and led to more robust behavioral restoration than did cell suspensions even when suspensions were supported with cografts of striatal cells or pretreatment with growth factors. PMID- 9739109 TI - Abrogation of betaglycan attenuates TGF-beta-mediated inhibition of embryonic murine lung branching morphogenesis in culture. AB - Although betaglycan (TGF-beta type III receptor) is known to enhance TGF-beta ligand binding to its type II receptor in murine lung epithelial cell lines, the biological significance of this phenomenon in the process of lung organogenesis is not understood. Betaglycan gene expression was detected in embryonic murine lungs undergoing branching morphogenesis in ex vivo culture. Antisense betaglycan oligodeoxynucleotides (ODN) resulted in up to 56% stimulation of lung branching morphogenesis in culture, while betaglycan mRNA and protein expression levels were suppressed by 90 and 82%, respectively. Following abrogation of betaglycan expression with antisense oligodeoxynucleotide, embryonic lungs were relatively insensitive to TGF-beta: TGF-beta2 (0.5 ng/ml) and TGF-beta1 (20 ng/ml), respectively, down-regulated lung morphogenesis by 38 and 34% in control cultures, whereas TGF-beta-induced inhibition was attenuated to 13 and 26% respectively, in the presence of betaglycan antisense oligodeoxynucleotides. TGF beta neutralizing antibodies also prevented TGF-beta-mediated inhibition of lung branching in culture, supporting the speculation that autocrine/paracrine TGF beta signaling is minimal in the absence of betaglycan. Betaglycan was immunolocalized mainly to the epithelial cells in developing airways, a spatial distribution which overlaps with that of TGF-beta type II receptor. Furthermore, abrogation of endogenous betaglycan gene expression prevented the characteristic down-regulation of cyclin A and surfactant protein C (SP-C) mRNAs by exogenous TGF-beta ligands. These results show that betaglycan expression is essential for optimal TGF-beta signaling during embryonic lung development. We therefore conclude that the abrogation of endogenous betaglycan attenuates endogenous autocrine and/or paracrine TGF-beta-mediated negative regulation of lung organogenesis. PMID- 9739110 TI - Reserpine attenuates D-amphetamine and MDMA-induced transmitter release in vivo: a consideration of dose, core temperature and dopamine synthesis. AB - Amphetamine releases dopamine through a transporter-mediated mechanism. The purpose of this report was to further our understanding of the intracellular pool from which amphetamine releases dopamine: the cytoplasmic pool, the vesicular pool, or both. Rats were treated with D-amphetamine (AMPH) (1.0 or 10.0 mg/kg) or an amphetamine analog, methylenedioxymethamphetamine (MDMA) (2.0, 5.0, or 10.0 mg/kg). Pre-treatment with 10.0 mg/kg reserpine (18 h prior to AMPH or MDMA) attenuated dopamine release for high and low AMPH doses; however the low-dose effect showed borderline significance. Pre-treatment with 10.0 mg/kg reserpine attenuated dopamine and serotonin release induced by MDMA. The dopamine effect was seen at all three MDMA doses; the effect on serotonin was only measured at the 10.0 mg/kg dose. Reserpine pre-treatment caused reductions in core body temperature; heating the rats to normal body temperature for 3 h prior to AMPH or MDMA, and during the 4 h post-treatment period partially reversed the reserpine induced attenuation of dopamine release. However, the intermediate level of dopamine release for the reserpinized-heated animals was not significantly different from either the reserpine group (not heated) or the AMPH or MDMA alone groups. In a separate group of rats, the effects of reserpine and reserpine+heat on dopamine synthesis were measured. DOPA accumulation after treatment with the aromatic acid decarboxylase inhibitor NSD-1015 (100 mg/kg, 30 min before sacrifice), was greater in rats treated with reserpine compared to controls; heating the reserpinized rats did not significantly alter the amount of DOPA accumulation; however there was a trend towards further increase. These results suggest that D-amphetamine releases dopamine that is stored in both vesicles and the cytoplasm. Cooling may contribute to the attenuation of AMPH or MDMA-induced dopamine release observed after reserpine; however, AMPH or MDMA dependence upon vesicular stores most likely explains the diminished release after reserpine. The attenuation of AMPH or MDMA-induced transmitter release by reserpine is thought to be counteracted by a reserpine-induced replenishment of stores. Therefore, all doses of D-amphetamine may use vesicular stores; the degree to which new synthesis counteracts the vesicular depletion may be the variable which differentiates low from high doses of D-amphetamine. PMID- 9739111 TI - A new factor derived from 1321N1 human astrocytoma cells causes differentiation of PC-12 cells mediated through mitogen-activated protein kinase cascade. AB - Glial cells play an important role in maintaining neural function. In the present study, we examined the effects of a factor derived from human astrocytoma cells (1321N1) on differentiation of rat pheochromocytoma cells (PC-12). The conditioned medium which had been used for culture of 1321N1 cells caused the differentiation of PC-12 cells, suggesting that 1321N1 cells release a neurotrophic factor. The factor was apparently distinct from well-known neurotrophic factors, such as nerve growth factor (NGF), since it was resistant to boiling and trypsin treatment. The molecular size of the factor was assumed to be below 1000 through dialysis and ultrafiltration experiments. Furthermore, PC 12 cells were differentiated synergistically by the combined addition of NGF and the conditioned medium of 1321N1 cells. Partially purified fraction of the factor by Sephadex G-15 gel filtration column caused the prolonged activation of mitogen activated protein kinase (MAPK). The differentiation of PC-12 cells induced by the fraction or NGF disappeared after the treatment with PD98059, a specific inhibitor of MAPK kinase (MEK), suggesting the involvement of MAPK in the differentiation. These results suggest that the new low-molecular factor derived from glial cells causes differentiation of PC-12 cells mediated through an activation of MAPK. PMID- 9739112 TI - Contribution of METRO pathway localized molecules to the organization of the germ cell lineage. AB - To elucidate the potential role of localized components in the specification of the germ cell lineage we analyzed the composition of the germ plasm in Xenopus laevis oocytes and early embryos with respect to the vegetally-localized RNAs. We focused on Xlsirts, Xcat2, and Xwnt11 transcripts that are localized to the vegetal cortex through a region of the mitochondrial cloud called the messenger transport organizer (METRO) that also contains the nuage or germ plasm. At the ultrastructural level Xcat2 mRNA was detected on germinal granules while Xlsirts and Xwnt11 were associated with a fibrillar network of the germ plasm in stage-1 and stage-4 oocytes. In embryos, we found that all three RNAs remained associated with the germ plasm. Vg1 mRNA, a transcript localized through the late pathway, was excluded from the germ plasm in oocytes and embryos. Addtionally, we detected the protein spectrin within 16 cell nests of germ cells, in a structure reminiscent of the Drosophila spectrosome. Spectrin was detected in the mitochondrial cloud and was found in the germ plasm during embryogenesis. These data indicate that the various RNAs found within METRO and the protein spectrin are integral components of the Xenopus germ plasm with the RNAs being associated with different subcellular structures. They also suggest that the pathway through which RNAs are localized during oogenesis may be an important factor in biasing their distribution into specific cell lineages. The presence of Xwnt11 in the germ cell lineage suggests that a wnt-directed signaling pathway may be involved in germ cell specification. differentiation or migration. PMID- 9739114 TI - XTrR-I is a TGFbeta receptor and overexpression of truncated form of the receptor inhibits axis formation and dorsalising activity. AB - We have previously cloned a type I serine/threonine kinase receptor from Xenopus, namely XTrR-I. We show here that XTrR-I is able to bind and mediate the activity of TGFbeta1, but is unable to mediate response to activin or BMP-4. We have made a truncated receptor construct that can act as a dominant negative mutant receptor, and this can block the activity of TGFbeta2 but not that of activin. Overexpression of either the full-length or truncated receptor has a drastic effect on mesoderm differentiation. The truncated receptor inhibits expression of notochord and muscle in mesodermalised animal caps, while the full-length receptor greatly increases the amount of notochord. In addition, the truncated receptor blocks the axis duplicating activity of both siamois and Xwnt8. We conclude that XTrR-I is involved in mediating a dorsalising activity important for mesoderm differentiation. PMID- 9739113 TI - The effect of neurointermediate lobe denervation on hypothalamic neuroendocrine dopaminergic neurons. AB - The contribution of tuberohypophyseal and periventricular-hypophyseal dopaminergic neurons to the regulation of the secretion of prolactin (PRL) has yet to be clarified. In this study, we used pituitary stalk compression to disrupt hypothalamic neural input to the neurointermediate lobe (NIL). Neurointermediate lobe denervation (NIL-D) selectively disrupts the axons of tuberohypophyseal and periventricular-hypophyseal dopaminergic neurons, while leaving tuberoinfundibular dopaminergic neurons and the vascular supply of the pituitary gland intact. NIL-D was performed in ovariectomized (OVX) rats. The concentration of DA and 3,4-dihydroxyphenylacetic acid (DOPAC) in the median eminence (ME) and various regions of the pituitary gland of OVX and OVX+NIL-D rats were measured by HPLC-EC. The concentration of PRL, alpha-melanocyte stimulating hormone (alpha-MSH), and luteinizing hormone (LH) in serum were determined by radioimmunoassay. Successful NIL-D was confirmed by increased water intake. One week after NIL-D, serum PRL and alpha-MSH were elevated, but there was no change in the concentration of LH in serum. The concentration of DA was increased in the median eminence (ME), decreased in the outer zone of the anterior lobe (AL-OZ), as well as the intermediate (IL) and neural lobes (NL), and remained unchanged in the inner zone of the anterior lobe (AL-IZ). The concentration of DOPAC was increased in the ME and NL, decreased in the IL, and remained unchanged in both the AL-IZ and AL-OZ. These data confirm that pituitary stalk compression denervates the NIL. Moreover, decreases in the concentration of DA in the IL and AL-OZ, coupled with elevation of serum PRL and alpha-MSH indicate that DA from the NIL contributes to the increased inhibition of the secretion of PRL and alpha-MSH in OVX rats. PMID- 9739115 TI - Effect of urocortin on ACTH secretion from rat anterior pituitary in vitro and in vivo: comparison with corticotropin-releasing hormone. AB - Both urocortin (UCN) and corticotropin-releasing hormone (CRH) are known to stimulate secretion of adrenocorticotropic hormone (ACTH) by corticotroph cells via type-1 corticotropin-releasing hormone receptor (CRHR-1). We extensively examined UCN effects on the anterior pituitary (AP), particularly on proopiomelanocortin (POMC) mRNA and CRHR-1 mRNA as well as ACTH secretion in vivo. Moreover, signal transduction with UCN exposure was assessed in AP cell cultures in comparison with transduction following CRH exposure. Intravenously administered of UCN (5 microg/kg) increased ACTH and corticosterone secretion. Similarly, intravenous administration of UCN increased POMC mRNA and decreased CRHR-1 mRNA in the AP. These UCN effects were more potent and long-lasting than those of CRH. The prominent effect of UCN on ACTH secretion in vivo was confirmed in AP cell cultures, where application of UCN stimulated ACTH release approximately 7 times more strongly than CRH. The effect of UCN on ACTH release was enhanced by phorbol esters which activate protein kinase C, but was reduced by the selective cAMP-dependent protein kinase inhibitor, H-89. These results suggest that, as with CRH, UCN stimulates ACTH production and/or release through cAMP-dependent mechanisms, and that protein kinase C-dependent mechanism has a synergistic effect upon UCN-induced ACTH release. The more potent effects of UCN relative to CRH may be attributable to UCN's higher affinity for CRHR-1. PMID- 9739116 TI - Increase in gonadotropin-releasing hormone (GnRH) levels in CSF after stimulation of the nervus terminalis in Atlantic stingray, Dasyatis sabina. AB - The nervus terminalis (NT) contains many cells immunoreactive to gonadotropin releasing hormone (GnRH). The potential of the NT to release GnRH in vivo was investigated by stimulating the peripheral nerve trunk of Atlantic stingrays and collecting cerebrospinal fluid (CSF). The CSF samples from stimulated animals averaged about twice the levels of mGnRH-like peptide as those of unstimulated animals. These results demonstrate that nervus terminalis activity can effect in vivo GnRH levels in the brain. PMID- 9739117 TI - Alteration of limb and brain patterning in early mouse embryos by ultrasound guided injection of Shh-expressing cells. AB - A basic limitation of the study of development in the mouse is the inaccessibility of the embryos, which are encased in the maternal uterus. We demonstrate the first use of ultrasound backscatter microscopy for guiding injections of cells and other agents into early stage mouse embryos. Cells were injected into the mouse neural tube cavity as early as 9.5 days post coitus (E9.5), and into the developing limb buds as early as E10.5. Furthermore, a cell line engineered to express the secreted factor Sonic Hedgehog (Shh) was injected into early developing mouse brains or limbs. The Shh-expressing cells were found to induce ectopic expression of the Shh target genes Patched and Hnf3beta in the dorsal brain, and to alter digit patterning in the anterior limb bud. These results show that gene misexpression studies can be performed in mouse embryos using ultrasound-guided injection of transfected cells or retroviruses. In combination with the many available mouse mutants, this method offers a new approach for analyzing genetic interactions through gain-of-function studies performed in mutant mouse backgrounds. PMID- 9739118 TI - Enhanced localization of amyloid beta precursor protein in the rat hippocampus following long-term adrenalectomy. AB - Using various antibodies to the amyloid ss precursor protein (APP) associated with Alzheimer's disease, we investigated changes in the distribution of APP in the hippocampus and neocortex of adrenalectomized (ADX) rats. In contrast to sham operated controls, ADX rats euthanised after a survival period of 5 months showed striking APP reactivity in the CA1-CA4 fields and in the surviving cells in the dentate gyrus. Our results suggest the enhanced APP reactivity in hippocampal neurons may pertain to previous observations on the accumulation of APP fragments in the neocortex during ischemic or traumatic injury. Thus, long-term hormone deprivation would be another factor, which may influence the expression of APP in brain. PMID- 9739119 TI - Microglia, but not astrocytes, react to sciatic nerve injury in aging rats. AB - Peripheral nerve axotomy activates microglia and astrocytes within regions of brainstem or spinal cord from which the nerve arises. The present study demonstrates that unilateral sciatic axotomy in rats 2 to 18 months of age results in differing responses with age between these two glial populations. By 4 days postaxotomy, both astrocytes and microglia become activated in 2-month-old rats, whereas only the microglial population shows evidence of activation in rats 8 to 18 months of age. PMID- 9739120 TI - Effects of palatability-induced hyperphagia and food restriction on mRNA levels of neuropeptide-Y in the arcuate nucleus. AB - This study examined the effect of feeding either a bland cornstarch-based diet (BCD) or a highly palatable, high fat diet containing sucrose (HPD) on hypothalamic arcuate nucleus (ARC) gene expression for neuropeptide-Y (NPY). Male Sprague-Dawley rats received either BCD ad libitum, HPD ad libitum, HPD pair-fed to the caloric intake of the BCD, or the HPD at 60% of ad libitum HPD intake for 7 days. Animals receiving the HPD ad libitum consumed more calories and gained more weight than animals receiving the BCD (P<0.001). The HPD did not affect ARC NPY mRNA levels, whether the subjects were allowed to overeat or pair-fed to the BCD (P>0.05). However, feeding the HPD at 60% of ad libitum intake of the HPD, increased NPY mRNA levels in the ARC relative to the other treatments (P<0.01). The present data are consistent with the view that NPY in ARC responds to energy deficits rather than to hyperphagia stimuli related to palatability. PMID- 9739121 TI - Mesodermal cell fate decisions in Drosophila are under the control of the lineage genes numb, Notch, and sanpodo. AB - In Drosophila, much has been learned about the specification of neuronal cell fates but little is known about the lineage of mesodermal cells with different developmental fates. Initially in development, individual mesodermal precursor cells are singled out to become the founder cells for specific muscles. The selection of muscle founder cells is thought to employ a Notch-mediated process of lateral inhibition, similar to what is observed for the specification of neural precursors. These muscle founder cells then seem to fuse with the surrounding, uncommitted myocytes inducing the formation of muscle fiber syncytia. In contrast, the differentiated progeny of neural precursor cells are usually the result of a fixed pattern of asymmetric cell divisions which are directed, in part, by interactions between numb, a localized intracellular receptor protein, sanpodo (spdo), a potential tropomodulin homolog, and Notch, a transmembrane receptor protein. Here, we have investigated the role of these neural lineage genes in the cell fate specification of muscle and heart precursors. In particular, we have focused on a progenitor cell that is likely to produce a mixed lineage, generating both a pericardial heart cell and a somatic muscle founder cell. We show that the asymmetric segregation of Numb into one of these daughter cells antagonizes the function of Notch and spdo by preventing the presumptive muscle founder from assuming the same fate as its cardiac sibling. Our results suggest that asymmetric cell divisions, in addition to the previously documented inductive mechanisms, play a major role in cardiac and somatic muscle patterning and that additionally the cytoskeleton may have a role in the asymmetrical localization of cell fate determinants. PMID- 9739122 TI - Propofol modulation of [3H]flunitrazepam binding to GABAA receptors in guinea pig cerebral cortex. AB - Binding of the radioligand [3H]flunitrazepam to membranes prepared from the cerebral cortex of adult, male guinea pigs under equilibrium and non-equilibrium conditions was used to investigate the allosteric interaction between the intravenous general anesthetic propofol and the benzodiazepine site of the GABAA receptor. Propofol induced a potentiation of [3H]flunitrazepam binding with an EC50 of 9+/-4 microM. Propofol increased the affinity for [3H]flunitrazepam binding with no change in maximal binding. Propofol did not change the rate constant of association for [3H]flunitrazepam binding to cerebral cortical membranes. In contrast, the rate constant for dissociation of [3H]flunitrazepam was significantly decreased in the presence of propofol. These data demonstrate that propofol increases the affinity of the benzodiazepine site of the GABAA receptor via a selective decrease in the rate constant for dissociation, and suggest a mechanism for the allosteric interaction between propofol and benzodiazepines at the GABAA receptor. PMID- 9739123 TI - Corrigendum to 'Melatonin inhibits spontaneous and VIP-induced vasopressin release from suprachiasmatic neurons' PMID- 9739124 TI - The specification of the pronephric tubules and duct in Xenopus laevis. AB - We have examined the timing of specification of the pronephric tubules and duct in Xenopus laevis by explanting the presumptive pronephric rudiments into blastula ectodermal wraps. We have established the time point of specification using the monoclonal antibody markers 3G8 and 4A6 which recognize antigens in pronephric tubule and duct, respectively. We show that, by experimental analysis in explants, kidney tubules are specified by stage 12.5 in the pronephric anlagen whereas pronephric duct is specified later between stages 13 and 14. Furthermore we show that signals involved in tubulogenesis of the pronephric tubules are normally received between stage 12.5 and 13. These experiments unambiguously pinpoint the timing of pronephros specification analyzed by explant experimentation to a developmental stage prior to that demonstrated for urodele amphibia, and provide an essential biological backdrop to a search for the molecular nature of pronephric inducers. PMID- 9739125 TI - Origin of noradrenergic afferents to the shell subregion of the nucleus accumbens: anterograde and retrograde tract-tracing studies in the rat. AB - The nucleus accumbens (NAcc) can be subdivided into 'core' and 'shell' based on anatomical connections and histochemical markers. Previous studies have demonstrated dopamine-beta-hydroxylase immunoreactive (DBH-ir) fibers in the NAcc shell, but the source of these noradrenergic (NE) afferents has not been determined. Therefore, we have investigated in detail the anatomy of NE afferents to this subregion. Dual immunohistochemistry for DBH and substance P demonstrated numerous DBH-ir fibers in the caudal NAcc shell. Neurons projecting to the NAcc were identified with Fluoro-Gold (FG) or cholera toxin B (CTb) retrograde tracing and tyrosine hydroxylase (TH) immunohistochemistry. Single- and double-labeled neurons were observed in the A2 and A1 NE cell groups following FG injections into the caudal NAcc shell. Numerous FG and CTb single-labeled neurons were found in the rostral locus coeruleus (LC), subcoeruleus and pericoerulear dendritic region, with an occasional double-labeled neuron in the LC. Few labeled neurons were seen in the brainstem after FG injections into the NAcc core, consistent with the lack of DBH-ir in this subterritory. To confirm these results, injections of Phaseolus vulgaris leucoagglutinin or biotinylated dextran amine were made into the LC or nucleus tractus solitarius (NTS). Virtually no labeled fibers were observed in the NAcc following injections into central LC. However, fibers were observed in the NAcc shell after injections in the NTS. These results indicate that the primary source(s) of NE afferents to the NAcc shell is the A2 region of the NTS, with lesser contributions from A1 and LC. PMID- 9739127 TI - Vacuolar neuritic dystrophy in aged mouse superior cervical sympathetic ganglia is strain-specific. AB - We have developed a model of autonomic nervous system aging using the mouse superior cervical sympathetic ganglion (SCG) which is characterized by the reproducible development of distinctive, markedly-enlarged neuritic swellings (vacuolar neuritic dystrophy, VND). These structures contained an admixture of lucent vacuoles and subcellular organelles, and involved both presynaptic and postsynaptic ganglionic elements. Quantitation of the frequency of VND was accomplished at the light microscopic level and validated by ultrastructural examination. VND lesions were 30-100-fold more frequent in the aged mouse paravertebral SCG than in the prevertebral celiac/superior mesenteric (C/SMG) sympathetic ganglia. Although VND was identified in all ages of mice examined, the number of lesions increased significantly with age. The frequency of VND was a function of the strain of mouse examined with a 40-fold difference in VND frequency between C57BL6 mice, the least involved strain, and the DBA/2J strain, which was most affected and began to develop significant numbers of lesions at an early age. As in our human studies of aging in the sympathetic nervous system, there was a prominent gender effect with males developing twofold greater numbers of VND lesions than females. Mice maintained on a significant calorie restricted diet for 30 months developed 70% fewer lesions than ad libitum-fed, age and sex matched controls. The aging mouse SCG, therefore, represents a robust animal model with reproducible, quantifiable and unambiguous neuropathology. Insights into pathogenetic mechanisms gained in the subsequent analysis of this relatively simple peripheral sympathetic nervous system model may contribute to the understanding of some of the most complex and significant problems involving higher brain function. PMID- 9739128 TI - Novel pattern of Brachyury gene expression in hemichordate embryos. AB - Together with echinoderms and chordates, hemichordates constitute the third major group of the deuterostomes, which share a number of common developmental features. The Brachyury gene is responsible for the formation of notochord, the most defining feature of chordates. Therefore, isolation and characterization of the hemichordate homolog of Brachyury is key to understand the origin and evolution of chordates. Here we show that the hemichordate Brachyury gene (PfBra) is expressed in two regions of the gastrula and young tornaria larva, the archenteron invagination region and the stomodeum invagination region. PMID- 9739130 TI - The expression patterns of endothelin-A receptor and endothelin 1 in the avian embryo. AB - We investigated the expression pattern of the endothelin-A receptor and endothelin 1 genes, the mutations of which affect the development of the mesectodermal derivatives of the neural crest. We show here that endothelin 1 is expressed by the environment of the cephalic neural crest cells invading branchial arches. Later on, while the neural crest-derived tissues of the head continue to express endothelin-A receptor, endothelin 1 is no longer expressed in their environment. PMID- 9739129 TI - Correlation between potentiation of AP1 DNA binding and expression of c-Fos in association with phosphorylation of CREB at serine133 in thalamus of gerbils with ischemia. AB - Protein biosynthesis is mainly under the control at the level of gene transcription in eukaryotes. Transcription factors are nuclear proteins with abilities to modulate the activity of RNA polymerase II which is responsible for the formation of messenger RNA from double stranded DNA in the cell nuclei. Binding of a radiolabeled oligonucleotide probe for the transcription factor activator protein-1 (AP1) was transiently potentiated 1 to 6 h after the recirculation of blood supply in the thalamus and striatum, but not in the entorhinal cortex, olfactory bulb, frontal cortex, cerebellar cortex and medulla pons, in gerbils with transient global forebrain ischemia for 5 min, in addition to the hippocampal subregions. The ischemic insult not only increased the immunoreactivity with an antibody against cyclic AMP response element binding protein (CREB) phosphorylated at serine133, but also induced the expression of both c-Jun and c-Fos family proteins 3 h after the recirculation in the thalamus. Limited proteolysis by Staphylococcus aureus (S. aureus) V8 protease revealed the expression of different partner proteins of AP1 in response to ischemic signals in the thalamus. Moreover, ischemia for 2 min led to more prolonged elevation of AP1 binding in the thalamus at least up to 12 h after the reperfusion than that seen with ischemia for 5 min. These results suggest that potentiation of AP1 DNA binding may at least in part involve mechanisms associated with the expression of c-Fos protein through phosphorylation of CREB at serine133 in the thalamus of gerbils with ischemia. PMID- 9739131 TI - Differential expression of the HMG box transcription factors XTcf-3 and XLef-1 during early xenopus development. AB - The recent discovery that the HMG box transcription factor XTCF-3 is involved in early axis specification in Xenopus laevis (Molenaar, M., van de Wetering, M., Oosterwegel, M., Peterson-Maduro, J. Godsave, S., Korinek, V., Roose, J., Destree, O., Clevers, H., 1996. XTcf-3 transcription factor mediates beta-catenin induced axis formation in Xenopus embryos. Cell 86, 391-399) led us to search for other members of the TCF/LEF family in this species. A newly identified HMG box factor was cloned with highest homology to human LEF-1, called XLEF-1. Unlike XTcf-3, XLef-1 is not expressed maternally, but its transcripts become detectable directly after the mid blastula transition (MBT). At later stages, both genes are expressed in the central nervous system (CNS), eyes, otic vesicles, head mesenchyme, neural crest and derivatives, branchial arches, developing heart, tailbud and limb buds. The expression pattern of Lef-1 during later stages of development is evolutionarily conserved. PMID- 9739132 TI - Hypoxic adaptation of the peptidergic innervation in the rat carotid body. AB - The abundance of substance P (SP)-, calcitonin gene-related peptide (CGRP)-, vasoactive intestinal polypeptide (VIP)-, and neuropeptide Y (NPY)-immunoreactive nerve fibers in the carotid body was compared between normoxic and chronically hypoxic rats (10% O2 and 3.0-4.0% CO2 for 3 months). The immunoreactive fibers appeared as thin processes with many varicosities, and were distributed mainly around the vasculatures. In the normoxic control carotid body, NPY fibers were more numerous than VIP, CGRP, and SP fibers. In the chronically hypoxic rats, the carotid body was enlarged several fold, and the mean absolute number of VIP and NPY fibers was 3.88 and 2.22 times higher than in the normoxic carotid body, respectively, although that of SP and CGRP fibers was not changed. When expressed as density per unit area of the parenchyma, the density of SP and CGRP fibers in the chronically hypoxic carotid body decreased significantly to under 50%, the density of VIP fibers increased significantly 1.80 times, and the density of NPY fibers were unchanged. Immunoreactivity for four neuropeptides was not found in the glomus cells of normoxic or chronically hypoxic carotid bodies. These results suggest that altered peptidergic innervation of the chronically hypoxic carotid body is one feature of hypoxic adaptation. Because these neuropeptides are vasoactive in nature, altered carotid body circulation may contribute to modulation of the chemosensory mechanisms by chronic hypoxia. PMID- 9739133 TI - Xenopus eomesodermin is expressed in neural differentiation. AB - Our initial description of the Xenopus gene Eomesodermin (Eomes) indicated that it is expressed largely if not entirely in mesodermal cells of gastrula stage embryos. A more detailed examination, described here, shows that it continues to be expressed in the most anterior (future head) mesoderm during gastrula and neurula stages. However, during tail-bud stages, Eomes expression is re-born in the most anterior part of the brain, becoming strongly transcribed in the olfactory region of the telencephalon. This later Eomes expression marks a very localized region of the forebrain distinct from that of Otx-2, anterior to that of En-1 and overlapping that of Sox-3. PMID- 9739134 TI - saliva, a new Drosophila gene expressed in the embryonic salivary glands with homologues in plants and vertebrates. AB - saliva (slv) transcription begins at the salivary gland placodes and continues on throughout development as salivary glands invaginate and reach their final location and morphology. saliva is located cytogenetically in 76A/B, and encodes a 226-amino-acid protein with four hydrophobic domains. A Northern blot detects a 1.6-kb transcript throughout development. Database similarity searches reveal homology to proteins from Caenorhabditis, Lilium, Medicago and mouse. PMID- 9739135 TI - CMIX, a paired-type homeobox gene expressed before and during formation of the avian primitive streak. AB - We cloned a chicken homeobox gene closely related to the Xenopus Mix. 1 gene. CMIX is expressed early in embryogenis in a sickle-shaped area in the posterior zone of the blastoderm. With the beginning of gastrulation, CMIX transcripts are found in the primitive streak primordium, then in the young and medium-sized streak, however not in the mesoderm after its emergence. In the fully-extended streak, CMIX is restricted to its middle, i.e. the prospective ventral mesoderm. CMIX RNA is undetectable by whole-mount in-situ analysis in later stages. We compare CMIX expression to the early pattern of the brachyury gene. PMID- 9739136 TI - Spatial and temporal expression of c-Fos protein in the spinal cord of anesthetized rat induced by subcutaneous bee venom injection. AB - In order to study central neuronal components involved in subcutaneous (s.c.) bee venom-induced persistent pain (a new tonic pain model), we use Fos immunostaining technique to study the spatial and temporal patterns of neuronal activity in the spinal cord of anesthetized rats. Following intraplantar bee venom injection, Fos like immunoreactive (ir) neurons were only seen from L1 to S3 rostrocaudally with distinct distribution at L4-5 segments. At segments of L1-2 and S1-3, Fos-ir labelings were diffusely and symmetrically distributed on both sides of the gray matter; however, at L4-5 segments, Fos-ir neurons were densely localized in medial portion of laminae I-II, less densely in laminae V-VI and a few in laminae VII and X ipsilateral to the injection side. No Fos labeling was seen in ventral horn of the spinal cord at L4-5 segments. Fos protein began to express only within lamina I at 0.5 h, but increased over the whole dorsal horn at 1 h and reached peak labeling at 2 h after bee venom. Expression of c-Fos in laminae I-II decreased at 4 h, and completely disappeared at 24 h, however, labeling in laminae V-VI disappeared much slowly and existed even at 96 h after bee venom. Within laminae III-IV, Fos-ir neurons could not be seen at 0.5 h, but began to be seen at 1 h and appeared to exist even at 24 h after bee venom. Systemic morphine suppressed c-Fos expression dose-dependently in both superficial and deep layers of dorsal horn and the latter region was much more sensitive to morphine than the former one. The present results demonstrated that prolonged neuronal activities in superficial and deep layers of dorsal horn were essential to mediation of bee venom induced tonic pain and may have different roles in generation and/or modulation of spontaneous pain and hyperalgesia and allodynia. PMID- 9739137 TI - Characterization of CMIX, a chicken homeobox gene related to the Xenopus gene mix.1. AB - Members of the TGFbeta, Wnt and FGF families act in concert to induce and pattern the mesoderm of gastrulating embryos. Downstream effectors for these growth factors include homeobox proteins, which also feed back to activate and repress upstream signaling pathways (e.g. Fainsod, A., Steinbeisser, H., De Robertis, E.M. 1994. On the function of BMP-4 in patterning the marginal zone of the Xenopus embryo. EMBO J. 13, 5015-5025; Carnac, G., Kodjabachian, L., Gurdon, J.B., Lemaire, P. 1996. The homeobox gene Siamois is a target of the Wnt dorsalization pathway and triggers organizer activity in the absence of mesoderm. Development 122, 3055-3065). As well as having interwoven upstream and downstream regulatory pathways Mix.1, siamois and goosecoid, all paired-type homeobox genes, may physically interact with each other as heterodimers to regulate dorsal ventral polarity (Mead, P.E., Brivanlou, I.H., Kelley, C.M., Zon, L.I. 1996. BMP 4 responsive regulation of dorsal-ventral patterning by the homeobox protein Mix.1. Nature 382, 357-360). We report here a chicken paired-type homeobox gene, CMIX, with a homeodomain having 72% aa identity to its nearest homolog, Xenopus Mix.1. CMIX is expressed in the epiblast of the posterior marginal zone of early chick embryos, and later along the entire anterior-posterior axis of the primitive streak in cells of the medial ectoderm, in nascent mesoderm, but not in endoderm. Coincident with formation of prechordal mesoderm, CMIX mRNA levels rapidly decline throughout the embryo. PMID- 9739138 TI - Xenopus cadherin-11 is expressed in different populations of migrating neural crest cells. AB - We cloned the Xenopus homologue of cadherin-11 and studied its spatiotemporal expression pattern during early development. The messenger RNA is present from the mid-gastrulation through embryo development. It is expressed in different neural crest cell populations, during their migration and differentiation. This pattern, unexpected for an adhesion molecule, reinforces the idea of novel functions for type II cadherins. PMID- 9739139 TI - Effects of applied currents on spontaneous epileptiform activity induced by low calcium in the rat hippocampus. AB - It is known that both applied and endogenous electrical fields can modulate neuronal activity. In this study, we have demonstrated that anodic current injections can inhibit spontaneous epileptiform events in the absence of synaptic transmission. Activity was induced with low-Ca2+ (0.2 mM) artificial cerebrospinal fluid (ACSF) and detected with a voltage threshold detector. At the onset of an event, a current was injected into the stratum pyramidale via a tungsten electrode positioned within 150 micron of the recording site. Data was recorded with a glass pipette electrode. The results show that spontaneous epileptiform activity can be fully suppressed by subthreshold anodic currents with an average amplitude of 3.9 microA and a minimum amplitude of 1 microA. In addition, we observed that some events could be blocked by current pulses with shorter durations than the duration of the event itself. The possibility that increased tissue resistance could contribute to the efficacy of the currents was tested by measuring the step-potential increase evoked by anodic current injections. The data show a significant increase in the amplitude of the evoked potential after introduction of a low-Ca2+ medium, suggesting that tissue resistance is increasing. These results indicate that low-amplitude, subthreshold current pulses are sufficient to block epileptiform activity in a low-Ca2+ environment. The increased tissue resistance induced by sustained exposure to a low-Ca2+ medium could contribute to the low current amplitudes required to block the epileptiform events. PMID- 9739140 TI - Impairment of long-term potentiation and paired-pulse facilitation in rat hippocampal dentate gyrus following developmental lead exposure in vivo. AB - Neonatal rats were exposed to lead from parturition to weaning via the milk of dams drinking 0.2% lead acetate solution. The alterations of long-term potentiation (LTP) and paired-pulse facilitation (PPF) of hippocampal dentate gyrus in adult rats (90-115 days) following developmental lead exposure were studied in vivo. Input/output (I/O) function, paired-pulse facilitation (PPF), excitatory postsynaptic potential (EPSP) and population spike (PS) amplitude were measured in the dentate gyrus (DG) in response to stimulation applied to the lateral perforant path. The results showed that LTP was induced in control rats with an average PS potentiation of 321.1+/-50.0% (n=18), which was significantly greater than the increase in PS potentiation (173.5+/-30.0%, n=17, p<0.001) in lead-exposed rats after tetanizing stimulation. The mean EPSP potentiation increased to 172.4+/-27.0% (n=18) in control and 138.8+/-21.4% (n=17) in lead exposed rats after tetanizing stimulation. The lead-induced impairment of LTP of PS potentiation was more serious than that of EPSP potentiation. Following pairs stimulation of perforant fiber at 250 microA and an interpulse interval (IPI) of 10-1000 ms, the average peak facilitation of PS was 211.3+/-25.0% (n=13) in control and 187.7+/-23.0% (n=11) in lead-exposed rats. The average facilitation period duration of PS was 243.0+/-35.8 ms (n=13) in control and 138.0+/-24.4 ms (n=11) in lead-exposed rats. These results suggested that developmental lead exposure in neonatal rats caused impairments in LTP and PPF of hippocampal dentate gyrus. PMID- 9739141 TI - Agonists of A1 and A2A adenosine receptors attenuate methamphetamine-induced overflow of dopamine in rat striatum. AB - The effect of adenosine receptor agonists on the release of striatal dopamine (DA), induced by repeated doses of methamphetamine (MTH), was evaluated. Rats received three injections of MTH (5 mg/kg i.p.) at 2-h intervals. The release of DA in the striatum was measured by a microdialysis in freely moving animals. The agonist of adenosine A1 receptor, N6-cyclopentyladenosine (CPA) and the agonist of adenosine A2A receptor, 2-[p-(carboxy-ethyl)phenylethylamino]-5'-N ethylcarboxyamidoade nosine (CGS 21680), either of them being infused locally into the striatum at concentrations of 50 and 100 microM, produced decreases in the extracellular DA level during exposure to MTH, and a weaker effect on the levels of DOPAC and HVA. The above effects were reversed by the specific antagonists of adenosine A1 and A2A receptors, 8-cyclopentyl-1,3-dipropylxanthine (DPCPX) and 3, 7-dimethyl-1-propargylxanthine (DMPX), respectively. Our results indicate that both the A1 and A2A adenosine receptors appear to be involved in reducing the excessive release of DA in the striatum; furthermore, they suggest a neuroprotective role of adenosine in MTH neurotoxicity. PMID- 9739142 TI - Monoclonal antibody monospecific to glycine for brain immunocytochemistry. AB - We have developed mouse monoclonal antibodies (AGLY-1-8, all IgG1 subisotype mAbs) against glycine (Gly) conjugated to bovine serum albumin using glutaraldehyde (GA)-NaBH4. Among these, AGLY-4 mAb was found to be the most useful for Gly immunocytochemistry (ICC) in functions of specificity and sensitivity without non-specific immunobinding. AGLY-4 was demonstrated to be monospecific to Gly by an enzyme-linked immunosorbent assay (ELISA) binding test, and not reactive to any of the other amino acids and peptides tested. Using this antibody, indirect immunoperoxidase staining was observed in different regions of the rat brain fixed with GA in combination with borohydride reduction. In contrast, immunoreactivity was quite low in tissues fixed only with GA. Absorption controls indicated that the immunostaining could be completely inhibited by 5 microg/ml of Gly-human serum albumin (HSA) conjugate prepared using GA and NaBH4, which was consistent with the results of an ELISA inhibition test. No cross-reaction occurred with other GA-conjugated amino acids. Dense ICC staining was observed in the rat neurons related to the auditory and vestibular centers, and modest immunostaining was seen in all the structures of the cerebellar cortex except for the Golgi cells which were strongly stained. These results were in complete agreement with the previous methods using polyclonal anti-Gly serum. Also, a new finding was that staining was noticed in certain cells widely distributed in the different brain regions. These results strongly suggest that the monoclonal antibody has a potential for elucidating the precise distribution of Gly-containing cells. PMID- 9739143 TI - Efferent connections of the lamina terminalis, the preoptic area and the insular cortex to submandibular and sublingual gland of the rat traced with pseudorabies virus. AB - Neurones situated in the lamina terminalis (organum vasculosum of the lamina terminalis, median preoptic nucleus and subfornical organ) as well as within medial and lateral parts of the preoptic area and in the insular cortex become transneuronally labelled following pseudorabies virus injections into the submandibular or the sublingual gland. These neurones are efferently connected to a chain of central neurones directed to secretory or vascular tissue of the submandibular or the sublingual gland. By varying the postinoculation time a stepwise infection of different forebrain nuclei was registered, with the hypothalamic paraventricular nucleus and the lateral hypothalamic area being the first forebrain structures labelled. Such early infected neurones within these hypothalamic nuclei are in all likelihood third order neurones regulating salivary secretion and might have functioned as relays transmitting virus to other forebrain structures. The above mentioned forebrain areas together with several other hypothalamic nuclei as well as the bed nucleus of the stria terminalis, the central nucleus of the amygdala and the substantia innominata, seem to be the widespread anatomical basis for the central regulation of salivary gland function. PMID- 9739144 TI - Genetic differences in morphine sensitivity, tolerance and withdrawal in rats. AB - Significant genetic differences in the endogenous opioid system and in response to a variety of noxious stimuli are present in rodents. We now compared the response to noxious heat with the hot plate test, morphine sensitivity and the development of tolerance and dependence to morphine in spontaneously hypertensive (SHR), Wistar-Kyoto (WK) and Sprague-Dawley (SD) rats. Significant differences were observed in basal nociception among the three strains, where SHRs were hypoalgesic compared to WK and SD. The antinociceptive effect of morphine varied among strains (SD>SHR>WK) as did the rate of tolerance development (10 mg/kg morphine 2/day for 4 days) where WK>SD=SHR. SHR rats developed hyperalgesia following morphine administration during the course of tolerance development. Furthermore, although naloxone (2 mg/kg) precipitated withdrawal symptoms in all tolerant rats, the panorama of symptoms varied among the three strains. Thus, there are significant genetic differences in a variety of effect of opiates. PMID- 9739145 TI - Physical training modifies the age-related decrease of GAP-43 and synaptophysin in the hippocampal formation in C57BL/6J mouse. AB - We investigated the effect of a moderate amount of prolonged physical training initiated at 3 months of age on the expression of GAP-43 and synaptophysin in the hippocampal formation. C57BL/6J mice were divided into three groups which were trained (24 months old), sedentary (24 months old) and young (3 months old). From 3 months of age on, mice of trained group were treated with voluntary running wheel for 1 h each day (5 days per week) until 24 months of age (21 months running), whereas mice of sedentary group were put in immobilized wheels for the same time. Using immunohistochemistry and image analysis system, GAP-43 and synaptophysin were analysed quantitatively in the CA1, CA3 areas and the dentate gyrus of the hippocampal formation. As compared with young mice, the densities of GAP-43 and synaptophysin immunostaining showed a significant decrease in the hippocampal formation in sedentary group (P<0.01). After 21 months of running, the densities of GAP-43 and synaptophysin immunostaining significantly increased in the examined areas of the hippocampal formation in trained mice compared to their age-matched sedentary controls (P<0.05, 0.01). These results indicate that a moderate amount of prolonged physical training could modify the age-related decrease of the expression of GAP-43 and synaptophysin in the hippocampal formation, and that the increased expression of GAP-43 and synaptophysin might be associated with the anatomical sprouting and synaptogenesis. PMID- 9739146 TI - Chloride concentration in cultured hippocampal neurons increases during long-term exposure to ammonia through enhanced expression of an anion exchanger. AB - The effects of long-term exposure to ammonia on [Cl-]i in cultured hippocampal neurons were examined. Ammonia increased the [Cl-]i time- (>/=24 h) and concentration- (>/=2 mM) dependently, resulting in a depolarizing shift of the equilibrium potential of the GABAA receptor-Cl- channel opening (EGABA). Such an effect of ammonia was diminished by the inhibitors of Cl-/HCO3- exchangers, 0.1 mM 4-acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic acid (SITS) and 0.1 mM 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS), and a carbonic anhydrase inhibitor, 2 mM acetazolamide, but not by a Na+/K+/2Cl-cotransport inhibitor, 50 microM bumetanide, suggesting an enhanced Cl-/HCO3- exchange activity by ammonia. The ammonia-induced increase in [Cl-]i was also abolished by the inhibitors of protein kinase C (PKC), 0.1 microM calphostin C and 10 microM 1 (5-isoquinolinyl-sulfonyl)-2-methylpiperazine dihydrochloride (H-7), and of transcription and de novo protein synthesis, 1 microM actinomycin D and 0.5 microg/ml cycloheximide, while a PKC activator, 0.1 h microM phorbor 12-myristate 13-acetate (PMA), increased the [Cl-]i. The mRNA level of the AE3 Cl-/HCO3- exchanger was increased by ammonia in a calphostin C- and H-7-sensitive manner. The AE3-like immunoreactivity was also increased by ammonia. These findings suggest that long-term exposure to ammonia increases the expression of AE3 through the activation of PKC, resulting in an increase in [Cl-]i in neurons and a reduction of inhibitory postsynaptic potentials. PMID- 9739147 TI - Serotonin 5-HT2c agonists mimic the effect of light pulses on circadian rhythms. AB - The serotonin agonist quipazine has been shown to cause phase shifts in melatonin and activity rhythms and to induce c-fos in the suprachiasmatic nucleus of rats. In this study, in vivo pharmacological characterisation of the phase shifting properties of serotonin agonists has been performed, with a view to determining the receptor sub-types involved. Agonists for the 5-HT2a/2c receptors, (+/-)-1-(4 iodo-2,5-dimethoxyphenyl)-2-aminopropane hydrochloride (DOI, 0.1 mg/k), 1-(3 chlorophenyl)-piperazine HCl (mCPP, 2 mg/kg) and N-(3-trifluoromethylphenyl) piperazine HCl (TFMPP, 2 mg/kg) injected at CT18 resulted in acute transient inhibition of melatonin production and delays in the onset of production on the following nights of 1.2+/-0.2, 1.7+/-0.3 and 1. 4+/-0.8 h respectively. Drugs specific for 5-HT1a/7 and 5-HT3 receptors failed to affect melatonin production. At a dose of 0.07 micromole/kg, the serotonin antagonist, ritanserin inhibited the DOI induced phase delay whereas ketanserin was ineffective at this dose, providing strong evidence that DOI was acting through 5-HT2c receptors. DOI (0.5 mg/kg) at CT18 provoked a phase delay in the core body temperature rhythm of similar magnitude to that following a light pulse. Administration of DOI but not agonists active at other receptor sites resulted in the appearance of c-Fos in the ventrolateral division of the suprachiasmatic nucleus (SCN) at CT18 but not at CT6. Ritanserin was more potent than ketanserin at inhibiting the DOI induced increase in c-Fos labelled cells in the SCN. When rats were pre-treated with metergoline (15 mg/kg), ritanserin (3 mg/kg) or LY 53,857 (3 mg/kg) prior to a 2 lx/ 1 min light pulse, none of the drugs significantly inhibited the responses to light. The results of these experiments indicate that serotonergic agonists active at the 5-HT2c receptor mimic the effects of light on 2 independent rhythms and activate SCN neurones in the rat. PMID- 9739148 TI - Genetic polymorphism of paraoxonase 1 (PON1) and susceptibility to Parkinson's disease. AB - Toxicologists have thought that the paraoxonase (PON) enzyme polymorphism might contribute to effects of pollutants and other environmental chemicals on susceptibility to cancer, birth defects and Parkinson's disease (PD). We studied a biallelic PON1 polymorphism at codon 192 (A and B alleles) in 166 patients with sporadic idiopathic PD. The frequency of the B (Arg) allele of PON1 was significantly increased in patients with PD than in healthy controls (chi2=8.75, df=1, P<0.005). The relative risk of PD in homozygotes for the B allele was 1.60 fold higher than individuals with the A (Gln) allele (chi2=7.38, df=1, P<0.01). Our data suggest that environmental neurotoxins metabolized by PON1 might be responsible for neurodegeneration with aging and that the B (Arg) allele form might have genetic susceptibility to PD. PMID- 9739149 TI - Assembling an actin cytoskeleton for cell attachment and movement. PMID- 9739150 TI - Transport of fragmented DNA in apical dendrites of gerbil CA1 pyramidal neurons following transient forebrain ischemia. AB - Transport of fragmented DNA in apical dendrites of the CA1 pyramidal neurons of gerbil hippocampus is observed in the apoptotic process following transient forebrain ischemia. The time-course of specific DNA fragmentation was examined after the ischemic insult by in situ nick-end-labeling method and fluorescence detection technique by DAPI. Although the role of the fragmented DNA movement is unclear, the transport mechanism of fragmented DNA is still active in the late phase of apoptotic process. PMID- 9739151 TI - Delayed recovery of auditory cortical evoked potentials is correlated with cortical neuronal death after transient cerebral ischemia in awake gerbils. AB - Early detection of irreversible neuronal change after transient cerebral ischemia is important so that adequate treatment can be initiated within the therapeutic window. We have examined the correlation between changes in middle-latency auditory evoked potentials (MAEPs) and histological changes in the auditory cortex of awake Mongolian gerbils subjected to 4 min or 12 min of transient cerebral ischemia. Post-ischemic MAEPs were characterized by the appearance of a markedly large negative and positive component at approximately 17-22 ms latency in both groups. Delay in the appearance of the high amplitude (maximal amplitude at 45 min after recirculation in the 12-min ischemia group) precedes the slowly developing death of neurons in the auditory cortex that results from transient cerebral ischemia. PMID- 9739152 TI - Brain NPY Y1 receptors rapidly mediate the behavioral response to novelty and a compartment-specific modulation of granulocyte function in blood and spleen. AB - Neuropeptide Y (NPY) alters behavioral activity and innate immune functions of rats within minutes of intracerebroventricular (i.c.v.) application. Using combinations of the Y1-5a,b(6) agonist NPY, the Y1,3,5 agonist [Leu31-Pro34]NPY (LP-NPY), and the selective Y1 antagonist BIBP3226 (BIBP), we investigated whether the NPY-Y1 receptor (Y1R) subtype regulates NPY-induced behavioral and immunological effects at 15 min after i.c.v. application. Administration of both NPY and LP-NPY decreased rearing activity in the open field and suppressed granulocyte function in the blood. These effects were blocked by BIBP pre treatment. In contrast to the blood, NPY and BIBP+NPY treatments stimulated granulocyte function within the splenic compartment. In addition, a blood leukophilia composed of granulocytes and NK cells was induced by NPY only. We conclude that the tested early effects of NPY are mediated by either the Y1R (rearing, blood granulocyte function), or a non-Y1R (splenic granulocyte function), or by a combined receptor activation (leukocyte mobilization). Furthermore, the immunological effects of NPY demonstrate compartment specificity. PMID- 9739153 TI - Nucleotide metabolizing ectoenzymes are upregulated in A431 cells periodically treated with cytostatic ATP leading to partial resistance without preventing apoptosis. AB - Extracellular ATP, when added as a single dose at concentrations higher than 0.1 mM to the culture medium, was growth inhibitory or even cytotoxic for human epidermoid carcinoma cells (A431). Adenosine at the same concentrations was much less potent. The molecular mechanism underlying the inhibitory effect of extracellular ATP has been investigated. The cytostatic as well as the cytotoxic effects of ATP could be prevented by supplying uridine as a pyrimidine source and, alternatively, by simultaneous addition of dipyridamole, which inhibits the uptake of adenosine. The data suggest that the long-term production and continuous uptake of adenosine, which is enzymatically generated from the ATP in the medium, led to an intracellular nucleotide imbalance with pyrimidine starvation. This triggered suicidal processes ending up in apoptosis of the cells. The tumor cells have been adapted to extracellular ATP with the aim to obtain cells which are more resistant to ATP. Therefore, growing cells were periodically treated with extracellular ATP. These cells were characterized by an enlargement of cell size, a decreased proliferation rate, and a reduced but not abolished sensitivity to cytostatic and cytotoxic ATP doses. The calcium response of adapted cells was shortened. The nucleotide hydrolyzing ectoenzyme activities (ecto-ATPase, ecto-ADPase, ecto-AMPase, ecto-Ap4Aase) were simultaneously upregulated. All phenotypic alterations of the adapted cells disappeared after cultivation for several generations in the absence of extracellular ATP. Considering ATP as a potential chemotherapeutic agent the adaptive phenomena of treated cells might be important. PMID- 9739154 TI - Nicotinic acetylcholine receptor mediated modulation of evoked excitatory amino acid release in the nucleus tractus solitarius of the rat: evidence from in vivo microdialysis. AB - In vivo microdialysis was used to measure release of endogenous l-glutamate and l aspartate in the nucleus tractus solitarius of the anaesthetised rat evoked by baroreceptor loading. Aortic constriction, the method of loading, elicited a reproducible increase in extracellular levels of l-glutamate to 322+/-139% of basal levels, which could be attenuated by concomitant local administration of the nicotinic acetylcholine receptor antagonist mecamylamine (100 microM). PMID- 9739155 TI - Opiate receptor avidity is reduced in non-motor impaired MPTP-lesioned rhesus monkeys. AB - Opiate receptor avidity, roughly equivalent to the ratio of unoccupied receptor density to the receptor dissociation constant (B'max/KD), was measured in four MPTP (1-methyl-4-phenyl-1,2,3, 6-tetrahydropyridine)-lesioned rhesus monkeys and nine normal controls with positron emission tomography (PET) and 6-deoxy-6-beta [18F]fluoronaltrexone (cyclofoxy, CF), a mu- and kappa-opiate receptor antagonist. Although the MPTP-lesioned monkeys were dopamine deficient as measured with [18F]-L-fluorodopa ([18F]-DOPA) and PET [Doudet et al., 6-[18F]-L DOPA imaging of the dopamine neostriatal system in normal and clinically normal MPTP-treated rhesus monkeys, Exp. Brain Res. 78 (1989) 69-80], they had clinically recovered from the acute motor effects of MPTP exposure. Opiate receptor avidity was found to be reduced by 30-35% in the opiate-receptor rich areas of caudate, anterior putamen, thalamus, and amygdala of the MPTP-lesioned animals. The results suggest that opiate pathways make a significant contribution to the adjustment of cortico-striatal-thalamic pathway activity and thereby to behavior in rhesus monkeys following dopamine loss. PMID- 9739157 TI - Suppression of cell adhesion and spreading activities of fibronectin by arginine specific ADP-ribosyltransferase from chicken polymorphonuclear leukocytes. AB - Arginine-specific ADP-ribosyltransferase present in secretory granules of chicken polymorphonuclear leukocytes (so-called heterophils) was shown to be released into the extracellular space by secretagogues (Terashima et al., J. Biochem. 120 (1996) 1209-1215). In the present work, we examined fibronectin as an extracellular target protein of the released transferase. Fibronectin was ADP ribosylated by purified transferase and stoichiometry of ADP-ribose incorporation into fibronectin was 1.0 mol/mol of fibronectin. Cell adhesion and spreading assays revealed that ADP-ribosylation of fibronectin markedly inhibited the adhesion activity of fibronectin. A proteolytic peptide map of ADP-ribosylated fibronectin demonstrated that the modification occurs in the cell binding domain of fibronectin. ADP-ribosylation of the RGD peptide suggests that the RGD sequence is the modification site in the domain. ADP-ribosylation of fibronectin in plasma means that fibronectin can probably serve as the substrate for extracellularly released ADP-ribosyltransferase in vivo. Thus, in the extracellular space, ADP-ribosyltransferase released from polymorphonuclear leukocytes may perhaps be involved in regulation of cell adhesion process by interfering with the activity of fibronectin. PMID- 9739159 TI - Caveat: mycoplasma arginine deiminase masquerading as nitric oxide synthase in cell cultures. AB - We used confluent cultures of dog gallbladder epithelial cells, stimulated by conditioned medium from a culture of human neonatal foreskin fibroblasts, to establish the presence of inducible nitric oxide synthase (NOS, EC 1.14.13.39). Assay was by conversion of radiolabeled arginine to citrulline. By 4 days after addition of the conditioned medium, a relatively high level of activity was observed. However, further study showed that the enzyme did not require addition of the usual cofactors for maximal activity (NADPH, FAD, FMN and tetrahydrobiopterin) and was stable in the absence of anti-proteolytic agents. Our suspicion that this enzyme might not be NOS but arginine deiminase (EC 3.5.3.6) was confirmed by enzyme purification and by the liberation of ammonia during enzyme reaction. This enzyme, which is absent from primates and virtually confined to single-cell organisms, suggested the presence of Mycoplasma, a common contaminant of cell cultures, and it was subsequently confirmed that the fibroblast culture was a source of Mycoplasma. With the widespread interest in nitric oxide and NOS, and common use of the convenient [3H]arginine assay, there is a considerable danger of the two enzymes being confused. At the very least, it is necessary to check for activity in the absence of added cofactors. PMID- 9739158 TI - MPP+ toxicity and plasma membrane dopamine transporter: study using cell lines expressing the wild-type and mutant rat dopamine transporters. AB - The Parkinsonism-inducing neurotoxin 1-methyl-4-phenylpyridinium (MPP+) causes specific cell death in dopaminergic neurons after accumulation by the dopamine transporter (DAT). COS cells, a non-neuronal cell line insensitive to high doses of MPP+, becomes sensitive to MPP+ when transfected with the rat DAT cDNA. We analyzed the bi-directional transport of MPP+ and its toxicity in several cell lines expressing wild or mutant DATs. Cell death in COS cells expressing wild DAT by exposure to MPP+ was concentration-dependent and cocaine-reversible. Increased wild DAT expression caused higher sensitivities to the toxin in HeLa cells. Although several mutant DATs demonstrated greater transport activity than the wild-type, they displayed similar or lower sensitivity to MPP+ toxicity. Reverse transport of preloaded [3H]MPP+ through DAT was facilitated in COS cells expressing certain mutant DATs, which consistently displayed less sensitivity to MPP+ toxicity. These results suggest that re-distribution of MPP+ due to influx/efflux turnover through the transporter is a key factor in MPP+ toxicity. PMID- 9739160 TI - Characterization of pI(Cln) binding proteins: identification of p17 and assessment of the role of acidic domains in mediating protein-protein interactions. AB - pICln is a ubiquitous and abundant 27 kDa soluble protein that is localized primarily to the cytoplasm. The protein has been proposed to be a swelling activated anion channel or a channel regulator. Recent studies, however, have cast significant doubt on these hypotheses, and the function of pI(Cln) therefore remains unknown. To further characterize the physiological role of pI(Cln), we have begun to identify the proteins that bind to it and the amino acid domains that mediate pICln protein-protein interactions. Using affinity assays and immunoprecipitation we have identified three proteins in C6 glioma cells with molecular masses of 17 kDa, 29 kDa and 72 kDa that bind selectively to pI(Cln). Microsequencing revealed that p17 is the non-muscle isoform of the alkali myosin light chain. pI(Cln) contains three acidic amino acid domains termed AD1, AD2 and AD3. Mutation of AD1 and/or AD2 had no effect on p17, p29 and p72 binding. However, binding of p72 was lost when four acidic amino acid residues were mutated in AD3, which is located at the carboxy terminus. A truncation peptide containing the last 29 amino acids of pI(Cln) was able to bind p72 normally. These results indicate that the carboxy terminus is necessary for p72-pI(Cln) interaction. Based on these and other findings, we propose that pI(Cln) is a protein responsible for regulating the structure and function of the cytoskeleton, and/or a protein involved in mediating interactions between components of intracellular signal transduction pathways. PMID- 9739162 TI - Ultrastructural localization and comparative distribution of nitric oxide synthase and N-methyl-D-aspartate receptors in the shell of the rat nucleus accumbens 1 PMID- 9739161 TI - Kininogen expression by rat vascular smooth muscle cells: stimulation by lipopolysaccharide and angiotensin II. AB - To identify the presence of a local kallikrein-kinin system in vascular wall, we have studied whether rat vascular smooth muscle cells (VSMC) express kininogen in vitro and in vivo. Western blots using anti-T-kininogen antibody revealed the presence of T-kininogen in conditioned medium of cultured VSMC. T-Kininogen secretion by VSMC was markedly enhanced by the addition of lipopolysaccharide (LPS), angiotensin II (AII) and phorbol 12-myristate 13-acetate (PMA) to the culture. Experiments using specific inhibitors for protein kinases and on the PMA induced down-regulation of protein kinase C suggested that a protein kinase C dependent or unidentified pathway is involved in AII or LPS action, respectively. The intravenous injection of LPS (0.5 mg/kg) resulted in an increase in T kininogen mRNA levels in the vascular smooth muscle of rat aorta, peaking at 16 h. Polyacrylamide gel electrophoresis of cDNA products generated by reverse transcription-polymerase chain reaction (RT-PCR) from aortic mRNA using primers specific for either T- or low-molecular-weight kininogen revealed that rat vascular smooth muscle expressed T-kininogen gene but not low-molecular-weight kininogen gene, and that LPS exclusively stimulated T-kininogen expression. The mRNA for high-molecular-weight kininogen was undetectable in either aortic smooth muscle or cultured VSMC by means of RT-PCR analysis. RT-PCR using specific primers for rat tissue kallikrein genes showed that aortic smooth muscle expressed KLK1 (true kallikrein) mRNA, but not KLK10 (T-kininogenase) mRNA. These results demonstrated that rat VSMC are a source of T-kininogen but not of low molecular-weight- or high-molecular-weight kininogen, in contrast to the expression of true kallikrein but not of T-kininogenase by these cells. PMID- 9739163 TI - Fusion of Sendai virus and individual host cells and inhibition of fusion by lipophosphoglycan measured with image correlation spectroscopy. AB - Fusion between Sendai virus (SV) and individual host cells was investigated with confocal laser scanning microscopy (CLSM) and image correlation spectroscopy (ICS). SV was labeled with the fluorescent probe 7-octadecylamino-4-nitrobenz-2 oxa-1,3-diazole (NBD-NH-C18) and was allowed to bind to host cells (HEp-2, BALB 3T3) at 4 degrees C. The effect of lipophosphoglycan (LPG), isolated from Leishmania donovani, on virus fusion was investigated by incorporation of LPG (0, 5, 10 or 20 microM) into the host cell membrane (HEp-2) before addition of SV. LPG did not affect the number of SV bound per cell. After incubation at 37 degrees C for 15 min without LPG, CLSM revealed a redistribution of NBD-NH-C18 from the SV envelope to the host cell membrane and an increase in average fluorescence intensity, indicating dequenching. ICS analysis of images obtained after incubation at 37 degrees C showed an increased mean cluster density to 260% of the value at 4 degrees C, reflecting the disappearance of labeled SV from the cell surface and diffusion of NBD-NH-C18 into the host cell membrane. Preincubation of the cells with LPG inhibited the temperature-induced redistribution and dequenching of NBD-NH-C18 in a concentration-dependent manner, with a total inhibition of fusion at 20 microM LPG. Together, the results demonstrate that CLSM combined with ICS is a powerful tool for studies of fusion of enveloped viruses with individual host cells and that LPG inhibits the fusion process at or before the hemifusion (lipid mixing) stage of SV interaction with cells. PMID- 9739164 TI - Degradation of distinct forms of multimeric vitronectin by human fibroblasts. AB - The plasma protein vitronectin is thought to be an important regulator of extravascular plasminogen activation. In previous studies we have shown that a disulfide stabilized multimeric form of vitronectin is endocytosed and degraded by fibroblast cells (T.S. Panetti, P.J. McKeown-Longo, J. Biol. Chem. 268 (1993) 11988-11993; P.J. McKeown-Longo, T.S. Panetti, in: K.T. Preissner, S. Rosenblatt, C. Kost, J. Wegerhoff, D.F. Mosher (Eds.), Biology of Vitronectins and their Receptors, Elsevier Science Publishers, Amsterdam, 1993, pp. 111-118). The preparation of multimeric vitronectin used in these earlier studies was in the form of high molecular weight disulfide-bonded aggregates which were stable in sodium dodecyl sulfate (SDS). To address the question of whether vitronectin needed to be in the form of disulfide stabilized multimers in order to be endocytosed, a multimeric vitronectin, which was not disulfide stabilized, was prepared from vitronectin that had been treated with reducing agent and alkylated with iodoacetamide. The resulting protein migrated as a 65/75 kDa protein on SDS gels in the absence of reducing agent, confirming that this form of vitronectin was no longer stabilized into disulfide-bonded aggregates. However, the protein was still multimeric when analyzed by native gels and could be converted to SDS stable multimers by cross-linking agents. This result demonstrated that reduced and alkylated vitronectin aggregates into multimeric forms which are not stable in SDS. Similar to disulfide stabilized multimers, alkylated multimers of vitronectin bound to sulfated proteoglycans in the extracellular matrix and were endocytosed and degraded. Degradation of both forms of vitronectin was inhibited with arginine-glycine-aspartic acid peptides, an anti-alphavbeta5 antibody and heparin. Chloroquine and wortmannin were also able to inhibit degradation of both forms of vitronectin, indicating that both multimeric forms were following the same endocytic and degradative pathway. These results suggest that the organization of vitronectin into a multimeric form which will be recognized for endocytosis does not require disulfide bond stabilization. This study further suggests that recognition of vitronectin for endocytosis is dependent upon its conversion from a monomeric to a multivalent form (C.E. Wilkins-Port, P.J. McKeown-Longo, Mol. Biol. Cell 8:S:64A (1997). PMID- 9739165 TI - Alterations in human lymphocyte DNA caused by sulfur mustard can be mitigated by selective inhibitors of poly(ADP-ribose) polymerase. AB - Changes in genomic DNA caused by exposure to the cytotoxic alkylating agent, 2,2' dichlorodiethyl sulfide (sulfur mustard; HD), alone or in combination with selective inhibitors of poly(ADP-ribose) polymerase (PARP), were analyzed as a function of HD concentration and post-exposure time. Preparations of human peripheral blood lymphocytes were exposed to HD (1x10(-8) M-1x10(-3) M), and incubated at 37 degrees C for 0-24 h. Total genomic DNA was extracted from these cells and compared with DNA from control cells of the same donor using agarose gel electrophoresis. The effects of HD on genomic DNA depended on the HD concentration and the length of the post-exposure time interval. DNA fragmentation was detected as early as 2 h after exposure to 3x10(-4) M HD, or at 24 h after exposure to 6x10(-6) M HD. The qualitative DNA pattern, as well as the extent of DNA fragmentation, changed with post-exposure time. Exposure to HD caused a time-dependent shift in the DNA cleavage pattern from an oligonucleosome sized 'DNA ladder' characteristic of apoptotic cell death, to a 'broad band' pattern characteristic of necrotic cell death. DNA fragmentation was not observed if cells were killed with heat or with Lewisite. Treatment of cells with selective PARP inhibitors consistently altered the DNA fragmentation caused by HD exposure. The inhibitors arrested DNA fragmentation at the DNA ladder stage. This effect only was observed if the PARP inhibitors were applied within 8 h of HD exposure. We conclude that early inhibition of PARP activity can induce a switch in the mechanism of cell death caused by HD. Such a switch may be useful therapeutically to convert a lytic, pro-inflammatory cell death that includes the disintegration of dying cells (necrosis), into a slower, programmed cell death that includes absorption of dying cells (apoptosis). PMID- 9739166 TI - Upregulation of Kupffer cell beta-adrenoceptors and cAMP levels during the late stage of sepsis. AB - Although a burst of immunoresponsiveness may occur during the early stage of sepsis, late sepsis is characterized by severe immunodepression. In addition, although studies have shown that stimulation of macrophage beta-adrenoceptors results in an increase in cAMP and an associated reduction in macrophage phagocytic activity, it remains unknown whether Kupffer cell beta-adrenoceptor characteristics and cAMP levels are altered during polymicrobial sepsis. To study this, Sprague-Dawley rats were subjected to sepsis by cecal ligation and puncture (CLP). At 5 h (i.e., the early stage of sepsis) or 20 h (late sepsis) after CLP or sham operation, the liver was perfused with collagenase solution and Kupffer cells were isolated. beta-Adrenoceptor characteristics of the isolated Kupffer cells were determined using [125I]iodopindolol, and basal levels of cAMP were measured by radioimmunoassay. The results indicate that while maximum binding capacity (Bmax) of Kupffer cell beta-adrenoceptors was not altered at 5 h, it increased significantly at 20 h after CLP. Similarly, basal levels of cAMP in Kupffer cells did not change at 5 h but increased markedly at 20 h after the onset of sepsis. In contrast, the dissociation constant (Kd, 1/affinity) of Kupffer cell beta-adrenoceptors was not significantly affected by sepsis at both 5 h and 20 h after CLP. Thus, upregulation of beta-adrenoceptors and increase in cAMP levels in Kupffer cells occur during the late stage of polymicrobial sepsis, and this may contribute to the depression of macrophage phagocytic function under such conditions. PMID- 9739167 TI - Import rate of the E1beta subunit of human branched chain alpha-ketoacid dehydrogenase is a limiting factor in the amount of complex formed in the mitochondria. AB - Components of the mitochondrial branched chain alpha-ketoacid dehydrogenase multienzyme complex are all encoded by nuclear genes. The functional complex is formed with a known stoichiometric relationship of subunits, but how they enter the mitochondria and form the complex is not defined. Although cytosolic precursors for several of the proteins have been identified, the requirements for import and processing have not been described. Here we demonstrate the similar requirements for in vitro import and processing of the three catalytic subunits unique the this complex. Import was not affected by the amount of endogenous BCKD within the mitochondria. No cooperativity or competition among the subunits for import was found when subunits were used in combination. The relative rates of entry are E1alpha>E2>/=E1beta, making E1beta the limiting component supporting previously reported observations. PMID- 9739168 TI - Quantitative assay by flow cytometry of the mitochondrial membrane potential in intact cells. AB - Mitochondrial membrane potential, in situ, is an important indicator of mitochondrial function and dysfunction. Because of recent interest in the role of mitochondria in signaling, cell injury and cell death, there is a need for a convenient, sensitive and accurate method for the measurement of the mitochondrial membrane potential, Deltapsim, in situ, in a heterogeneous cell population. We have adapted a flow cytometry method for the quantitative measurement of DeltaPsim which utilizes the lipophilic, cationic, fluorescent probe 3,3'-dihexyloxacarbocyanine iodide (DiOC6(3)). We developed a new protocol in which cells are equilibrated with very low dye concentrations (<1 nM). Only under these condition, the cell fluorescence appears to be correlated with the magnitude of DeltaPsim, as evident from the sensitivity of the fluorescence to low concentrations of uncouplers, ionophores and inhibitors of the mitochondrial proton pumps. The magnitude of the plasma membrane potential, DeltaPsip, also affects cell fluorescence, and a procedure that corrects for this effect is outlined. This method offers a distinct advantage over existing methods for estimation of Deltapsim by flow cytometry. PMID- 9739169 TI - DNA damage induces p21 protein expression by inhibiting ubiquitination in ML-1 cells. AB - We previously reported that deferoxamine, an iron chelating agent, induced p53 and cell accumulation in the G1 phase of ML-1 cells in the same way as the DNA damaging agent, etoposide. Etoposide treatment increased expression of the p21 gene, a cyclin kinase inhibitor, at both the mRNA and protein levels. However, deferoxamine treatment only increased the p21 mRNA level without the appearance of a detectable protein product. A substrate for cyclin kinase, pRB, was unphosphorylated by etoposide treatment, but remained unaffected by deferoxamine, indicating that p21 was functional after etoposide, but not after deferoxamine treatment. Therefore, in the present study, we investigated the involvement of the ubiquitin proteasome pathway in post-transcriptional regulation of p21. By the addition of lactacystin, a proteasome inhibitor, to deferoxamine treatment, the level of unubiquitinated p21 protein product was similar to that induced by etoposide treatment, and the ubiquitinated p21 bands became apparent. After etoposide treatment, the level of ubiquitinated p21 was diminished and a high level of unubiquitinated p21 expression was observed. We concluded that (1) efficient expression of p21 protein requires inhibition of the ubiquitin proteasome pathway, and (2) DNA damage inhibits the ubiquitination of p21. PMID- 9739170 TI - Are tyrosine phosphorylation of p125(FAK) and paxillin or the small GTP binding protein, rho, needed for CCK-stimulated pancreatic amylase secretion? AB - Studies of a possible role of tyrosine phosphorylation in the secretory process in rat pancreatic acinar cells provide conflicting conclusions. Recent studies show that tyrosine phosphorylation of the focal adhesion kinase, p125FAK and the cytoskeletal protein, paxillin, may mediate a number of cellular changes and this phosphorylation is dependent on the activation of the small GTP binding protein, p21Rho (Rho). In this work we have investigated the role of tyrosine phosphorylation of each of these proteins and of the activation of Rho in pancreatic enzyme secretion. Pretreatment with genistein, a tyrosine kinase inhibitor, decreased CCK-8-stimulated tyrosine phosphorylation of p125FAK and paxillin and CCK-8-stimulated amylase secretion by more than 60%, raising the possibility that tyrosine phosphorylation of these two proteins could be important in the ability of CCK-8 to stimulate amylase release. However, genistein did not alter the amylase release stimulated by TPA but inhibited TPA stimulated p125FAK and paxillin tyrosine phosphorylation by 70%. Pretreatment with C3 transferase, which specifically inactivates Rho, causes a decrease in CCK 8-induced maximal amylase release by 33%. Moreover, C3 transferase pretreatment causes a 48% and a 38% decrease in the tyrosine phosphorylation of p125FAK and paxillin by CCK-8, respectively. Pretreatment with different concentrations of cytochalasin D, an actin cytoskeleton assembly inhibitor, completely inhibited CCK-8-stimulated tyrosine phosphorylation of p125FAK and paxillin without having any effect on either the potency or efficacy of CCK-8 at stimulating amylase release. Furthermore, cytochalasin D completely inhibited TPA-stimulated tyrosine phosphorylation of both proteins without affecting TPA-stimulated amylase release. These results show that tyrosine phosphorylation of p125FAK and paxillin is not required for CCK-8 stimulation of enzyme secretion. However, our results suggest Rho is involved in the CCK-8 stimulation of amylase release by a parallel pathway to its involvement in the CCK-8-stimulated tyrosine phosphorylation of p125FAK and paxillin. PMID- 9739171 TI - Inhibition of macrophage migration inhibitory factor secretion from macrophages by vitamin E. AB - Macrophage migration inhibitory factor (MIF) was identified in rat peritoneal macrophages by Western blot analysis and its secretion into culture medium by enzyme-linked immunosorbent assay. We investigated the effect of vitamin E on MIF production in macrophages in response to phorbol 12-myristate-13-acetate (PMA), calcium ionophore A23187, and lipopolysaccharide (LPS). Intraperitoneal injections of vitamin E (5 mg per rat) for 6 successive days resulted in a significant increase of alpha-tocopherol content in peritoneal macrophages (478.3+/-90.7 ng/106 cells) compared with the control (1.5+/-0.5 ng/10(6) cells). For the control macrophages, MIF content of the medium (2.5x10(6) cells/18 ml) without stimulation was 2.27+/-0.20 ng/ml after 14 h culture, whereas stimulation with calcium ionophore A23187 (400 nM) and LPS (5.0 microg/ml) induced the elevation of MIF content to 3. 66+/-0.41 and 4.12+/-0.58 ng/ml, respectively. On the other hand, vitamin E-enriched macrophages without stimulation showed less MIF content (0.77+/-0.23 ng/ml) than the control. Similarly, the increase of MIF of vitamin E-treated macrophages was significantly suppressed after stimulation with calcium ionophore A23187 or LPS, compared with the control macrophages. From analysis of intracellular MIF content by Western blot, we found no alteration of intracellular MIF content of vitamin E-macrophages, in contrast to the decreased content of control stimulated-macrophages. Taken together, these results indicate that vitamin E may contribute to the regulation of immune responses through regulation of MIF secretion. PMID- 9739172 TI - Modification of sulfhydryls of the skeletal muscle calcium release channel by organic mercurial compounds alters Ca2+ affinity of regulatory Ca2+ sites in single channel recordings and [3H]ryanodine binding. AB - The actions of two organic mercurial compounds, 4-(chloromercuri)phenyl-sulfonic acid (4-CMPS) and p-chloromercuribenzoic acid (p-CMB) on the calcium release channel (ryanodine receptor) from rabbit skeletal muscle were determined by single channel recordings with the purified calcium release channel, radioligand binding to sarcoplasmic reticulum vesicles (HSR) and calcium release from HSR. p CMB or 4-CMPS (20-100 microM) increased the mean open probability (Po) of the calcium channel at subactivating (20 nM), maximally activating (20-100 microM and inhibitory (1-4 mM) Ca2+ concentrations, with no effect on unitary conductance. This activation was partly reversed by 2 mM DTT. Both compounds affected the channels only from the cytosolic side, but not from the trans side. 100 microM 4 CMPS caused a transient increase in Po, followed by a low activity state within 1 min. At inhibitory Ca2+ concentrations Po was increased to values observed with maximally activating Ca2+ or lower, inhibitory Ca2+ concentrations. The p-CMB/4 CMPS modified channels were ryanodine sensitive and blocked by ruthenium red. [3H]Ryanodine binding was increased up to four-fold with 3-15 microM 4-CMPS/p-CMB (Hill coefficient 1.7-2.0) at 4 microM Ca2+ and reduced at high concentrations (50-200 microM). The increase in [3H]ryanodine binding by 10 microM 4-CMPS was completely inhibited by 2 mM DTT. 4-CMPS significantly increased the affinity for the high affinity calcium activation sites and decreased the affinity of low affinity calcium inhibitory sites of specific [3H]ryanodine binding. 4-CMPS increased the affinity of the ryanodine receptor for high affinity ryanodine binding without a change in receptor density. 4-CMPS induced a rapid, concentration-dependent, biphasic calcium release from passively calcium-loaded HSR vesicles at subactivating Ca2+ concentrations (20 nM), which was partly inhibited by 4 mM DTT and completely blocked by 20 microM ruthenium red. It is suggested that the 4-CMPS-induced modulation of essential sulfhydryls involved in the gating of the calcium release channel results in a modulation of the apparent calcium affinity of the activating high affinity and inhibitory low affinity calcium binding sites of the calcium release channel. PMID- 9739173 TI - Effect of growth hormone on the translocation of GLUT4 and its relation to insulin-like and anti-insulin action. AB - To elucidate the effect of growth hormone (GH) on the insulin signal transduction pathway leading to the translocation of glucose transporter-4 (GLUT4), we constructed Chinese hamster ovary cells that overexpressed GH receptor and GLUT4. Treatment with GH triggered GLUT4 translocation, and this translocation was completely inhibited by wortmannin. GH-induced GLUT4 translocation reached a maximum level after 30 min, and then gradually decreased and returned to the basal level after 2 h. Tyrosine phosphorylation of JAK2 also became maximal after 30 min and then gradually decreased. In contrast, GLUT4 translocation remained unchanged for 2 h after insulin treatment, and tyrosine phosphorylation of insulin receptor substrate-1 (IRS-1) also remained constant for up to 2 h. Chronic GH treatment had almost no effect on insulin-stimulated Akt kinase activation and GLUT4 translocation. These results suggest that GH and insulin translocate GLUT4 in a similar manner, at least in part, and the difference in translocation depends on the difference in the tyrosine phosphorylation of JAK2 and IRS-1. The anti-insulin action of GH after chronic GH treatment does not appear to be mainly due to the inhibition of GLUT4 translocation. PMID- 9739174 TI - A monoclonal antibody to a multiphosphorylated, conformational epitope at the carboxy-terminus of p53. AB - Mutations of the gene encoding the tumor suppressor protein p53 are the most common molecular alterations of cancer cells found in about half of all human tumors. Mutations which cluster in well-defined hot spots change the structure of the protein thus affecting its ability to bind to DNA. Post-translational modifications, primarily phosphorylation, might also influence how p53 binds to DNA or folds to its active tetrameric form. However, the lack of appropriate biochemical markers to characterize the status of phosphorylation in different cell types and in cells at different stages of tumor progression has prohibited such investigations. To generate a sensitive and phosphorylation-specific monoclonal antibody (mAb), we chemically synthesized the C-terminal 23 amino acid stretch of human p53 in a double-phosphorylated form. The peptide 371-393, carrying phosphate groups on Ser378 and Ser392, was co-synthesized with a turn inducing spacer and peptide 31D, an immunodominant T-helper cell epitope in mice of the H-2k haplotype. After immunization and fusion of splenocytes with myeloma cells, a number of mAbs were obtained, from which mAb p53-18 emerged as a highly sensitive reagent. By enzyme-linked immunosorbent assay, p53-18, a mAb of the IgM isotype, recognized phosphorylated p53, expressed in insect cells infected with a recombinant baculovirus but not p53 expressed in Escherichia coli. Moreover, murine p53 from insect cells could be immune purified with mAb p53-18. Mass spectrometry following tryptic digestion of the purified protein and liquid chromatography of the fragments verified the presence of phosphate groups at both Ser375 and Ser389. From the corresponding human protein fragments, mAb p53-18 bound to the immunizing peptide phosphorylated on Ser378 and on Ser392, but failed to cross-react with the unphosphorylated peptide, or peptides phosphorylated individually on either Ser378 or Ser392. The binding to the unphosphorylated peptide could be restored, however, if the peptide conformation was stabilized to that of an alpha-helix. The immunogenic nature of the multiphosphorylated C-terminus of p53 is indicated by the finding that human sera, mostly from cancer patients, preferentially recognized the double phosphorylated peptide over the monophosphorylated or unphosphorylated analogs. Antibody p53-18 appears to be a highly useful biochemical marker to detect low levels of p53 protein in different tissues, and to be a key tool to characterize the phosphorylation status of the C-terminus of p53 protein originated from various sources. PMID- 9739175 TI - Extracellular Ca2+ regulates the respiratory burst of human neutrophils. AB - The role of extracellular calcium in the activation of respiratory burst in human neutrophils was studied by using the receptor agonist, N-formyl-methionyl-leucyl phenylalanine (fMLP), and the activator of protein kinase C phorbol myristate acetate (PMA). The level of intracellular free calcium was measured by using both cell suspensions and single cells in the presence and absence of extracellular calcium. The Ca2+-ATPase inhibitor, thapsigargin, was used to activate higher Ca2+ influx, while a novel calcium channel blocker, panax notoginseng saponins (PNGS) was used to block the Ca2+ entry from extracellular space during the responding period of cells. It was found that about two-thirds of the activation of respiratory burst initiated by the receptor agonist were attributed to the Ca2+ influx under normal physiological conditions. The higher Ca2+ influx resulted in tremendous enhancement of the intensity of respiratory burst initiated by fMLP and marked acceleration of the onset of the respiratory burst stimulated by PMA. It is evident that both intra- and extracellular Ca2+ are required for full activation of the respiratory burst of human neutrophils, and the Ca2+ influx from extracellular space plays an important role either in generation of reactive oxygen metabolites or in activation of protein kinase C. PMID- 9739176 TI - Fibroblast and epidermal growth factor receptor expression in Xenopus oocytes displays distinct calcium oscillatory patterns. AB - Electrophysiological study performed with the voltage clamp technique was used to examine the intracellular calcium pathway activated by tyrosine kinase receptor members. Three FGF receptors from Pleurodeles PR1, PR3, PR4, homologs to human receptors, and the human EGF receptor were expressed in Xenopus oocytes. Under FGF1, FGF2 and FGF4 stimulation, PR1 and PR3 display a one phase inward chloride calcium dependent current superimposed by sustained oscillations, whereas PR4 did not show any oscillations. These currents were dependent on intracellular calcium mobilisation, as the responses were reduced by caffeine (10 mM). Solely PR4 responses were affected by an extracellular calcium depleted solution suggesting the involvement of concomitant extracellular and intracellular calcium intervention in the calcium chloride current, whereas PR1 and PR3 did not. Under EGF stimulation, the EGF receptor elicits a two component inward current composed of an undelayed rapid transient dependent on intracellular calcium store recruitment followed by a second slower current dependent on calcium influx. The specific pattern and amplitude of the calcium oscillations induced by the combinatorial action of growth factors on their receptors could be relevant in numerous calcium dependent cell functions. PMID- 9739177 TI - [Prodromal period of the first episode of schizophrenia]. AB - The article is a review of the literature on the prodromal period of the first episode of schizophrenia. According to the literature, the first episode of schizophrenia is usually preceded by a few years of prodromal period including non-specific behavioural and affective disturbances as well as psychotic-like and prepsychotic symptoms. The concepts formulated so far concerning a definition of prodromal symptoms were discussed. The data were summarized on clinical characteristics of the prodromal period such as symptomatology, frequency of symptoms, dynamics of changes and the period of occurrence of the prodromal symptoms. Four hypothetical models of transformation of prodromal symptoms into psychotic process were described. Also, the importance of the prodromal symptoms of the first episode of schizophrenia for early diagnosis and therapy of schizophrenia was discussed with regard to currently performed programs of early interventions in schizophrenia. PMID- 9739178 TI - [Psychopathological profile of acute schizophrenic syndromes diagnosed according to ICD-10 and DSM-IV criteria]. AB - Diagnostic and symptomatological profiles of schizophrenic syndromes diagnosed according to ICD-10 and DSM-IV were compared. For this reason a group of patients fulfilling at least one of these sets of criteria was created and then diagnostic and symptomatological profile was compared between those who fulfilled the ICD-10 and those who fulfilled DSM-IV criteria. 105 inpatients hospitalized in acute phase of their first or one of consecutive episodes were included--102 of them had fulfilled ICD-10, and 90 DSM-IV criteria of schizophrenia. Diagnostic concordance between the two systems of criteria was high (83%). Differentiation of diagnostic profile (i.e. difference between frequency of fulfilling the specific requirements of ICD-10 or DSM-IV criteria) of the symptoms in these two groups was not significant, expert of 6-month criterion of duration of illness, which was significantly less frequently valid in ICD-10 syndromes group. A comparison of symptomatological profiles (i.e. frequency and intensity of symptoms) of schizophrenic syndromes diagnosed by ICD-10 or DSM-IV criteria and described by several rating scales (PANSS, SAPS/SANS, KOSS-S) did not show any significant differences. Results suggested that despite of different ways of defining the schizophrenic syndromes in both diagnostic systems, disorders manifested in the groups of patients created by means of them are very similar in psychopathological picture. This seems to be a significant change in comparison to more prominent differences contrasting the previous versions of the diagnostic systems (i.e. ICD-9 and DSM-III-R). PMID- 9739179 TI - [Schizophrenia, schizophrenia-like disorders and delusional disorders in patients with anorexia nervosa: literature review and report of 3 cases]. AB - The authors present the review of literature concerning schizophrenia, schizophrenia type and delusional disorders in patients with a lifetime diagnosis of anorexia nervosa (AN). The authors describe also 3 patients (2 cases of paranoid schizophrenia and 1 case of catatonic syndrome). The clinical features in all patients are discussed. In 1 patient the catatonic symptoms occurred within the context of AN, (perhaps due to metabolic disturbances) and in 2 other cases the psychotic features occurred after recovery from AN. The authors discuss the occurrence of psychotic features in AN, and the possible function of starvation and metabolic disturbances in their aetiology. PMID- 9739180 TI - [Family burden of schizophrenic patients with various forms of psychiatric care]. AB - Caregiver burden was evaluated among family members of 90 schizophrenic patients from hospital psychiatric ward, day hospital or from community psychiatry unit. Psychopathology was evaluated with the use of PANSS while family burden with the use of Tessler's scale which allowed to differentiate between objective and subjective burden regarding assistance to the subject and patient's supervision. Schizophrenic symptoms were more severe in hospitalized patients than among patients from day hospital or patients treated in the community. Family burden, both subjective and objective was more severe among family members of hospitalized patients. There was no difference in the severity of family burden among family members of patients from day-hospital or from community psychiatry unit. The severity of positive and general schizophrenic symptoms (PANSS) correlated positively with the lack of patient's acceptance by a family member as well as with the global subjective family burden and with the necessity of taking control over patient. There was a positive correlation between the severity of schizophrenic negative symptoms and subjective family burden (dimension: assistance to the patient) and the sum of objective family burden. PMID- 9739181 TI - [Cathamnestic long-term study in early schizophrenia. The assessment of psychological status and social functioning]. AB - Mental condition of 142 patents with a diagnosis of schizophrenia (according to ICD X:F 20) first admitted to Psychiatric Hospital at the age of 13-18 was assessed twice. The first assessment took place during their first hospitalization at Psychiatric Hospital, the second one--23 years later. PANSS, BPRS GAS Scale were used at the examination. Mean value of the exacerbation of symptoms and improvement rate were calculated for each group of symptoms: positive, negative and psychopathological. The interpretation of the links between selected factors and rates of improvement as well as the mean value of exacerbation of symptoms were obtained on PANSS Scale. The links between the life assessment index before the first hospitalization and a mean value of points on the PANSS Scale at the discharge from hospital and in the cathamnesis for positive and negative symptoms as well as i cathamnesis for psychopathological symptoms were proved. The obtained data confirm that after the first discharge from hospital more patients function better in life than after cathamnestic examination. PMID- 9739183 TI - [The study of the costs of schizophrenia]. AB - The present paper considers the problems of measurement of the costs of psychiatric services. Classification of costs' studies includes cost of illness, cost-benefit analysis, cost-effectiveness and cost-utility studies. Costs should be comprehensively measured. Randomised clinical trial is recommended. Information of costs should be integrated with information on patient outcomes. The paper discusses the research which tries to estimate the costs and benefits of community care, using naturalistic or random patient sample methods. PMID- 9739182 TI - [Subjective quality of life in patients with chronic schizophrenia and in healthy persons]. AB - The goal of this work was comparison of quality of life in chronic schizophrenic patients with that in healthy persons. As we had expected, the level of QoL in healthy persons was significantly higher than QoL in the examined patients. It also turned out that the level of everyday functioning in the group of healthy persons had certain impact on their subjective QoL. In the group of schizophrenic patients such relation was not detected. The patients' QoL was not influenced by their general level of functioning (measured on the GAS scale) or intensity of psychopathological symptoms either. The healthy persons examined by us expressed satisfaction with their emotional life, sense of freedom and responsibility, life in general and family relations significantly more often than the persons with schizophrenia. The patients admitted also that in everyday life they found it most difficult to exact their rights, make decisions, organize their daily schedule and ask for help. PMID- 9739184 TI - [Case management]. AB - The present paper discusses the theoretical background and present the practical use of the case management model of psychiatric care. Case management is a model of psychiatric care, in which the therapeutic effort is not restricted to biological and psychological functioning of the patient, but is also directed at reinforcement of the patient's environmental resources and social support. The basic principles of cases management are the community of care and the use of the patient-therapist relationship in the treatment. The patient is managed by one person responsible for all the issues relating to the patient. That person, called case-manager, directs the treatment in consultation with a psychiatrist, helps the patient in contacts with institutions and undertakes interventions in the patient's community. The paper discusses the obstacles in introducing the community care model based on case-management principles in Poland. PMID- 9739185 TI - [Association meaning of the emotionally aggressive words in a group of schizophrenic patients and in healthy controls. Semantic analysis]. AB - Associational responses from 128 schizophrenic people and 120 healthy ones were collected. Four words with aggressive emotional shade were given as the stimuli. Semantic analysis of stimulus words was made, which made it possible to obtain a range of content categories. The collected material was analysed then according to them. Significant differences in five content categories were discovered. Basing on these differences we can infer that associational meaning at schizophrenic people is in some way varied. It was also possible to draw some conclusions concerning emotional life of the examined people. PMID- 9739187 TI - [Psychosomatic disorders in the elderly]. PMID- 9739186 TI - [The study of regional cerebral blood flow in psychiatric disorders]. AB - Regional cerebral blood flow is a radiological technique which is safe, non invasive and relatively cheap. This method allows to assess changes in brain metabolism in patients with psychiatric pathology. The specific changes have been found in regional cerebral blood flow in patients with schizophrenia, affective disorder, obsessive-compulsive disorder as well as alcohol withdrawal and dependence. Advantages and disadvantages of this technique are discussed. PMID- 9739188 TI - [Geriatric psychosomatic disorders and psychotherapy of the elderly. Current concept and approaches to treatment]. AB - In a geriatric acute hospital, a minimum of 20% of the patients aged 60 or older fulfilled the case criteria for a psychogenic illness. Previously published epidemiological investigations had presented similar results. However, due to the limited theoretical training and treatment experience, psychotherapists continue to show extreme reluctance to formulate an indication for psychotherapy in elderly patients. The paper gives an overview of current concepts in development psychology for the second half of life drawing attention to the role of physical aging processes as an "organiser" of development. Sexuality in late life and approaches to dealing with chronic pain are presented as somatic-psychosomatic aspects of aging that challenge present old age stereotypes. Taking typical symptoms into consideration, a basis is established for the discussion of a differentiated therapy indication for elderly patients. Emphasis is on psychoanalytical and cognitive behavioural psychotherapy methods. Finally, perspectives of research are presented. PMID- 9739189 TI - [CIPA (Comprehensive Individualized Process Analysis)--a method for combining quantitative and qualitative individual case analysis]. AB - The main question of psychotherapy research nowadays is how psychotherapy works. Hence, interest focuses mainly on the process of psychotherapy. General change mechanisms as well as the therapeutic interaction are in the center of research interest. On the basis of process outcome findings (Orlinsky, Grawe, Parks 1994) and the schema theory of Grawe (1987) we developed a research instrument allowing the analysis of therapies from a theoretically and quantitative approaches. The research instrument (CIPA-Comprehensive Individualized Process Analysis) consist of three parts: The scales of the first part assess the general working mechanisms and the therapeutic relationship. The second part allows a rating of the patient's interactions inside and outside the therapy. In the third part the individual schemata are rated. The instrument and the research strategy are being illustrated by means of a selected therapy recorded completely on video tape. The results are interpreted on the basis of the individual schema structure as well as the therapy outcome. The possibility offered by the new instrument to combine quantitative and qualitative research strategies is discussed. PMID- 9739190 TI - [The Symptom Checklist-90-R (SCL-90-R) for presenting statistically and clinically significant psychotherapy outcome]. AB - After a long period of discussions on the efficacy of psychotherapy, there is still a lack of conventions for measuring change after psychological treatment. This paper first describes the concept of statistical and clinical significance of change. Using the SCL-90-R as a commonly administered instrument we then propose conventions and cut-off points for its global severity score (GSI) and change after therapy. A German standard population and several psychotherapy samples were aggregated to determine cut-off points and confidence intervals (reliable change indices) for statistically and clinically significant changes. Tingey et al. (1996) proposed the use of multiple clinical groups (inpatients and outpatients) aiming at a more realistic determination of "stepwise" changes. We examined this procedure with our data. Results show that it is not applicable in the German samples collected so far. Initial SCL-90-R scores in these groups did not differentiate sufficiently between inpatients and outpatients. Therefore, according to Jacobson and Truax (1991), moving from a "functional" to a "dysfunctional" population is still the criterion for a clinically significant change. PMID- 9739192 TI - [Pulmonary valve dilatation in tetralogy of Fallot]. PMID- 9739191 TI - [Wrist joint instability]. PMID- 9739193 TI - Cellular autoimmunity to retinal specific antigens in Behcet's disease. PMID- 9739194 TI - [Congenital or acquired human epididymal obstructions: consequences and etiological diagnosis]. PMID- 9739195 TI - [Psychosomatic approach to psoriasis: life events and psoriasis]. PMID- 9739196 TI - [Triglyceride composition of Tunisian olive oils]. PMID- 9739197 TI - [Sandifer syndrome]. PMID- 9739199 TI - [Amniotic band syndrome. Prenatal ultrasonography of a case]. PMID- 9739198 TI - [Simultaneous tachycardia]. PMID- 9739200 TI - [Management of an external pancreatic fistula after pancreatitis: a case report]. PMID- 9739201 TI - [Predudodenal portal vein: a case report]. PMID- 9739202 TI - [Type A behavior and coronary disease: update]. PMID- 9739203 TI - [Value and practical applications of cytogenetic in situ hybridization technique]. PMID- 9739204 TI - [Value of long term central venous catheters im medical oncology:prospective study of 102 cases]. PMID- 9739205 TI - [Hygiene adherence in the intensive care milieu]. PMID- 9739206 TI - [Epidemiology of hepatitis E in Tunisia]. PMID- 9739207 TI - [Treatment of perineal and genital gangrene with honey]. PMID- 9739208 TI - [Substernal goiters]. PMID- 9739209 TI - [Prenatal diagnosis of congenital toxoplasmosis: value of PCR test on amniotic fluid]. PMID- 9739210 TI - [Sneddon syndrome: a case report]. PMID- 9739211 TI - [Gastric tumors of smooth muscle origin. Three case reports]. PMID- 9739212 TI - [Cerebral trunk hematoma during eclampsia (A case report)]. PMID- 9739213 TI - [Traumatic peripheral vascular injuries]. PMID- 9739214 TI - [Traumatic peripheral vascular injuries]. AB - 2-4% of vascular injuries need operative reconstruction. In polytraumatized patients the rate is even 10%. Arterial vascular repair should precede venous reconstruction and orthopaedic stabilization due to limb threatening ischemia. Penetration or blunt vascular trauma result either in acute blood loss, ischemia or compartmental compression. Reperfusion syndrome leads to vital threat of patient. Clinical assessment, measurement of limb pressures using a Doppler device and use of duplex ultrasonography are reliable adjuncts in the rapid evaluation. Arteriography is rarely indicated and should be spared for patients with abnormal physical examination. Minimizing ischemia (6-8 h) is an important factor in maximizing limb salvage. Vascular repair include direct anastomosis or lateral suture repair mostly combined with primary shortening of the extremity. In most cases autogenous vein graft is required. Rethrombosis, arteriovenous fistula and pseudoaneurysms are possible complications. Stabilisation of the fracture has priority over vascular reconstruction. The initial steps to success are surgical debridement, adequate bony stabilization mostly by external fixation, revascularisation of vascular injury, immediate fascial decompression and early soft-tissue reconstruction. The best results are obtained when a multidisciplinary approach is used combining expertise in orthopedic surgery, vascular surgery and plastic surgery. PMID- 9739215 TI - [Significance of systemic inflammation in 1,278 trauma patients]. AB - The association between the increasing severity of systemic inflammatory response syndrome (SIRS) and the incidence of post-traumatic complications and mortality was retrospectively investigated in 1278 injured patients. Patients were divided into three groups according to their Injury Severity Score (ISS) (group A: ISS > or = 9 < or = 16 points (n = 626); group B: ISS > 16 < 40 points (n = 589); group C: ISS > or = 40 points (n = 63). SIRS was defined according to the criteria of the American Consensus Conference. The number of fulfilled criteria determined its severity: moderate SIRS: 2 criteria fulfilled, intermediate SIRS: 3 criteria fulfilled, severe SIRS: 4 criteria fulfilled. Additionally, acute respiratory distress syndrome (ARDS) was defined according to the Murray-Score and the multiple organ dysfunction syndrome (MODS) according to the Goris-Score. The incidence of SIRS was 42% in group A, 70% in group B and 100% in group C (p < 0.05). The severity of SIRS increased with severity of trauma. Moreover, 178 of all injured patients (14%) developed septic complications. In parallel to SIRS, the incidence of these septic complications correlated with the severity of trauma. The occurrence and severity of ARDS and MODS correlated with increased severity of SIRS and septic complications. Among patients without SIRS 15% developed ARDS and 21% MODS. In contrast, patients with severe SIRS and septic complications demonstrated ARDS in 99% and MODS in 97%. In these patients, no correlation was found between the ISS and the incidence of ARDS or MODS. There were also stepwise increases in mortality rates in the hierarchy from SIRS to septic shock. While 13 of patients with modest SIRS (5%) and 32 of patients with intermediate SIRS (13%) died, the mortality rate of patients with severe SIRS was 19% (P < 0.05). In addition, a significant correlation between the incidence of septic complications and mortality was found. Injured patients with sepsis died in 13%, those with severe sepsis in 23%, and patients with septic shock in 33% (p < 0.05). Thus, the increasing severity of SIRS was associated with the occurrence of posttraumatic ARDS, MODS, and mortality. Using the number of fulfilled SIRS criteria for classifying systemic inflammation, its severity may be predictive for posttraumatic complications and outcome of injured patients. PMID- 9739216 TI - [CPAP-augmented spontaneous respiration in thoracic trauma. An alternative to intubation]. AB - Intubation and Positive End Expiratory Pressure Ventilation (PEEP) is a well established therapeutic strategy for impaired lung function, particularly following blunt chest trauma. Complications of this regime are however also well known and pose the question why non-invasive forms of respiratory assistance such as Continuous Positive Airway Pressure (CPAP) have only gained minor popularity. In a prospective study, 30 patients who had suffered blunt chest trauma were treated with CPAP administered by mask. The regime consisted of continuous administration of CPAP by a face-mask, with gradually increasing periods of spontaneous breathing. Initially a FiO2 of 0.33 (range 0, 28-0, 33) proved necessary. The initial CPAP level was 7 mbar (range 5-8) with an (Assisted Spontaneous Breathing) ASB of 15 mbar (range 13-8). FiO2 and CPAP/ASB levels were subsequently gradually reduced until no longer necessary. In all patients intubation and ventilation was avoided by this regimen. The treatment was well accepted by all patients and common ventilation associated complications such as pneumonia did not occur. In comparison with the former standard method of treatment the average ICU stay was dramatically reduced, principally due to not having to gradually wean patients from ventilation and sedation. Other positive benefits include normal communication and feeding with active early mobilisation leading to faster recovery, both physical and psychological. We conclude that non invasive respiratory techniques should be used more frequently and recommend further studies are undertaken to define the indications. PMID- 9739217 TI - [Arthroscopic stabilization of anterior shoulder instability with bioresorbable tacks]. AB - We examined 38 patients with an arthroscopic bioabsorbable tack repair for anterior shoulder instability in a prospective evaluation. The mean follow-up was 22 months (range 12 to 33). The average age was 28.4 years (range 15 to 57), the operation was performed at average of 50 months (3 to 244 months) after injury. Assessment using the Rowe score revealed excellent results in 33 and good results in 3 patients. 1 patient had a fair result and 1 had a poor result. 26 should obtained full range of motion, 11 had minor (< 10 degrees) loss of external rotation, 1 experienced greater (< 20 degrees) loss of external rotation. 3 of the 38 patients (8%) had recurrent instability, 1 patient with 2 preceding operations and atraumatic and voluntary dislocation, respectively. The recurrence rate of arthroscopic Bankart repair with bioabsorbable tacks are comparable to open Bankart procedures. Success of the procedure depends on appropriate surgical technique and suitable selection of patients with unidirectional, posttraumatic, anterior instability who are found to have well-developed ligamentous tissue. PMID- 9739218 TI - [Retrograde intramedullary nailing of humeral fractures with new implants. Analysis of 120 consecutive cases]. AB - Since 1993, 120 fractures of the humerus were treated by retrograde unreamed nailing. Operations were performed on simple, complex, compound and pathological fractures of the proximal three quarter of the humerus. On the proximal humerus, displaced two-part-fractures and occasionally three- or four-part-fractures were stabilized. In 110 cases a prototype of an unreamed humeral nail with deployable fins for proximal locking was employed. In another ten cases the new solid interlocking nail of the AO/ASIF was used. The operative procedure, rehabilitation program, complications and functional and radiological results are presented. Retrograde nailing offers a high patient comfort and good functional results (Constant-Score on average 87% of the opposite side). Complications were nail migration (8.3%), instability (3.8%), nonunions (5.8%) and iatrogenic fractures (5.8%). Patients with high grade osteoporosis, small proximal fragments and poor compliance have an increased rate of complications. PMID- 9739219 TI - [Endoscopic versus limited open technique for replacement of the anterior cruciate ligament. 4 years outcome of a prospective study]. AB - ACL reconstruction with patellar tendon graft has become a standard procedure. The graft can be inserted either using two tunnels and a lateral femoral incision or with a femoral half tunnel drilled from the joint, thus avoiding the lateral incision. Advantages of the single-incision technique in the early rehabilitation period have been claimed. 40 patients with ACL deficiency were included in a prospective randomized trial comparing single and two-incision technique with a follow-up period of 4 years. Preoperative data did not show any significant difference between the two groups. At follow-up no difference were observed with respect to complications or he progress of rehabilitation. Evaluation after 1 and 4 years according to the IKDC form revealed good to excellent results in 70% of all patients. The Tegner-score increased significantly, however most patients did not regain their former activity level. ACL-reconstruction reduced anterior translation of the knee significantly at 6 months follow-up. However, we observed a slight increase of anterior translation after 1 and 4 years in both groups; stability was comparable in both groups at all time periods. We conclude that an arthroscopic single-incision technique has no advantage compared to a mini-open two-incision technique for ACL reconstruction with patellar tendon graft in terms of subjective or objective parameters. PMID- 9739221 TI - [Expert assessment in trauma surgery]. PMID- 9739220 TI - [The "posterior cervical line". A radiodiagnostic parameter of the cervical spine in children]. AB - The Posterior Cervical Line helps to differentiate pseudosubluxation from the true dislocation C 2/3 in children. But the line can also be used to recognize other pathologic conditions of the upper cervical spine in children, like atlanto axial sagittal instability or rotatory dislocation. PMID- 9739222 TI - [Internal osteosynthesis after unstable pelvic ring fracture in a 3-year-old child]. AB - A 3-year-old child was trapped under the heavy load of a forklift truck and sustained an unstable pelvic ring fracture (Tile type C) with complete SI disruption, disruption of the public symphysis and external rotation injury of the contralateral SI joint. An immediate internal fixation was performed, exposing the SI joint and the public symphysis simultaneously. For stabilization an H-plate was used for anterior plate fixation of the SI joint, while the public symphysis was stabilized by screws and cerclage wires. After one revision of the symphysis the clinical course was uneventful with anatomical healing of the pelvic ring. The implants were removed after 4 months. Clinical and radiological follow-up after 12 months showed no signs of maldevelopment of the pelvic ring. PMID- 9739223 TI - [Hospital planning]. PMID- 9739224 TI - Challenges of traction in critical care: a case study. AB - Traction can sometimes seem like an overwhelming challenge in the critical care setting not only to the patient and family, but also to the nurse. Critical care nurses must know the principles, purposes, methods, and patient care considerations necessary to be able to provide appropriate care and teaching for the patient in traction. This article describes these principles and then illustrates them through a case study. The case describes a patient who experiences multiple trauma and focuses on the nurse's role in the care of this patient in traction. PMID- 9739225 TI - Management of venous ulcers. AB - As our society ages, critical care nurses will increasingly be exposed to patients with a variety of wounds. One of the most challenging wounds to manage is the venous ulcer, a complication of venous insufficiency. This article will summarize the prevention and management of venous ulcers in a comprehensive and research-based manner. PMID- 9739226 TI - Orthopaedic infections. AB - Patients with orthopaedic infections may require admission to the intensive care unit (ICU). Necrotizing fasciitis and clostridial myonecrosis (gas gangrene) are serious soft tissue infections that may cause life-threatening complications. Patients suffering from infectious arthritis, osteomyelitis, or prosthetic joint infections may be seen in the ICU as a result of a previous injury, surgery, or delayed infectious processes. This article introduces the ICU nurse to the pathophysiology, clinical presentation, and management of a variety of orthopaedic infections. PMID- 9739227 TI - Clearing cervical spine injuries: a discussion of the process and the problems. AB - This article identifies and describes the standards of care in treating, diagnosing, and managing suspected cervical spine injuries. Staff experience, nursing, and medical literature provide insight into the complications associated with spine immobilization. Review of medical literature provides background on the standard diagnostic exams utilized and the rationale for the number and variety of tests included in the clearance process. Opportunities for reviewing practice, with an emphasis on limiting the effects of immobilization, can provide future challenges to nursing. PMID- 9739228 TI - Pediatric pain management in an adult critical care unit. AB - Many children every year will be treated in "adult" critical care units because of the limited pediatric trauma centers currently available. Assessment is an integral part of all pain management. Ideally, self-report is the gold standard for assessing pain; however, some children may not have the ability to use these tools. Nonverbal children may be assessed with behavioral tools such as the CHEOPS or FLACC. In children as young as 3 years old, the self-report tool called an OUCHER can be administered to assess their pain. Easy to apply nonpharmacological approaches are discussed with recommendations for nurses to incorporate into their daily care. PMID- 9739229 TI - Musculoskeletal transplant. AB - Allograft tissue has been used for more than two centuries in the repair of musculoskeletal injuries and defects. More than 150,000 procedures are performed each year in the United States alone, 10 times the number of organ transplants. Nurses caring for allograft recipients may be unfamiliar with where tissue comes from, how it is recovered, and the variety of its practical applications. Similar to the process of organ donation, with which many nurses have had experience, it all begins with the gift. PMID- 9739230 TI - Neuromuscular scoliosis: a case of the pediatric patient in the adult ICU. AB - Neuromuscular scoliosis can be a problem in children with underlying neuromuscular conditions such as cerebral palsy, spina bifida, and muscular dystrophy. A comprehensive preoperative assessment is essential to provide comprehensive postoperative care. Surgical procedures to correct neuromuscular scoliosis include anterior spinal fusion, posterior spinal fusion, or a combined anterior-posterior spinal fusion. Postoperative problems can include respiratory failure, hemodynamic instability, neurovascular compromise, and pain control. With an understanding of the developmental status of these patients, pediatric patients can be safely managed in an adult ICU. PMID- 9739232 TI - [Breast-feeding and breast cancer] PMID- 9739231 TI - Crush injuries: a case of entrapment. AB - Crush injuries result from prolonged entrapment or immobilization. The resultant complications can be devastating and fatal. There is significant tissue injury and muscle necrosis that may lead to one or more complications including compartment syndrome, rhabdomyolysis, renal failure, and coagulopathies. Using a case study format, this article discusses the chain of events surrounding crush injuries and the nursing management issues important for minimizing further complications for critically ill patients. PMID- 9739234 TI - [A small guide to scientific popularization] [In Process Citation] PMID- 9739233 TI - [The path of your texts. From your pen ... to the readers] [In Process Citation] PMID- 9739235 TI - [Economy at the service of society]. PMID- 9739236 TI - [An adolescent with suicidal ideas]. PMID- 9739237 TI - [Perinatal grief. To intervene with tact with the distressed parents]. PMID- 9739238 TI - [An experience in community health in Nicaragua. Call for solidarity]. PMID- 9739239 TI - [Child psychiatry. Promoting a positive self image]. PMID- 9739241 TI - [Milk and asthma: incompatible?] [In Process Citation] PMID- 9739240 TI - [Aphasic patients regain the pleasure of communicating]. PMID- 9739242 TI - [Mental health: tenacious prejudices]. PMID- 9739243 TI - [Mental health. How to prevent burnout?]. PMID- 9739244 TI - Pain relief for the dying. PMID- 9739245 TI - Rescuing Winnie the Pooh. PMID- 9739246 TI - Documenting suspected child abuse, Part I. PMID- 9739247 TI - Antimicrobial products: good infection fighters? PMID- 9739248 TI - Applying cardiac monitor electrodes. PMID- 9739249 TI - Reducing risks with bacteriostatic flush solutions. PMID- 9739250 TI - Whiplash. PMID- 9739251 TI - Is a nursing shortage on the way? PMID- 9739252 TI - Sizing up your patients for heparin therapy. PMID- 9739253 TI - Debunking six myths about enteral feeding. PMID- 9739254 TI - Sailing to Brittany. PMID- 9739255 TI - How to make the most of nonopioid analgesics. PMID- 9739256 TI - Taking orders by phone? PMID- 9739257 TI - Angina pectoris: a cry from the heart. PMID- 9739258 TI - Taming your anger. PMID- 9739259 TI - Less is more. PMID- 9739260 TI - Looking up to Patty. PMID- 9739261 TI - Hypoglycemia. Making a case for glucose gels and tablets. PMID- 9739262 TI - The nurse as patient advocate. PMID- 9739263 TI - Myths and facts... about epilepsy. PMID- 9739264 TI - Controlling postoperative nausea. PMID- 9739265 TI - Biliblanket phototherapy light. PMID- 9739266 TI - Refusing to believe. PMID- 9739267 TI - Health service history at a glance. PMID- 9739268 TI - Pages of history. PMID- 9739269 TI - My nursing life. PMID- 9739270 TI - The pioneers. PMID- 9739272 TI - The people's check-up. PMID- 9739271 TI - A study in hygiene and discipline? PMID- 9739273 TI - Foresight. PMID- 9739274 TI - Cultural influences on medical disclosure. AB - Personal freedom and individualism are highly valued in Anglo-American culture and consequently in healthcare. However, not all cultures recognize the value of these principles. Healthcare can be compromised when the patient's cultural context differs from that of the clinician. Culture and its influences on the disclosure of health-related information are described. Self-determination from an historical perspective, the role of the family, and a deeper understanding of paternalism as they relate to cultural issues are illustrated through a case study. Suggestions for creating a culturally relevant healthcare practice is presented. PMID- 9739276 TI - Pharmacology: impact on bladder function. AB - While numerous medications are effective in improving bladder function, others are implicated in the impairment of bladder function. The elderly patient in particular is vulnerable to a range of adverse effects from medications, including urinary incontinence. Information dealing with the basics of the interaction between voiding and drug receptors associated with the bladder is presented as background for the discussion of pharmacologic therapies for stress, urge, reflex, and overflow urinary incontinence. The most clinically significant medications in terms of effectiveness, dosage, pharmacokinetics, and side effects are noted with mention of drugs currently being researched. Over-the-counter, prescription and social drugs that may contribute to impaired bladder function are identified. Other nonpharmacologic therapies that may be used concomitantly with pharmacotherapy or in isolation are noted. PMID- 9739277 TI - The role of estrogen in female urinary incontinence and urogenital aging: a review. AB - Urogenital aging is a complex of urogenital symptoms involving the lower urinary tract, the genital tract and the pelvic floor. These symptoms involve hypoestrogenism in the menopausal woman. This review concludes that irritative urinary and local vaginal symptoms are quite amenable to estrogen therapy. Urinary incontinence is thought to be benefited by treatment with estrogen, although controversy exists. There is a limited role for estrogen in problems of urogenital prolapse, rectal symptoms, and sexuality in menopause. PMID- 9739278 TI - Post prostatectomy urinary incontinence. AB - The specific incidence rate of post prostatectomy incontinence is difficult to ascertain. However, regardless of the type of prostatectomy, whether it be transurethral, radical retropubic or radical perineal prostatectomy, or the nature of the prostatic disease, several risk factors are common to all. The most significant risk factors include pre-existing detrusor and/or sphincter dysfunction, increasing age, and surgical expertise. Management options include behavioral techniques, pharmacologic therapy, surgical intervention, and other supportive measures. While no definitive preventive measures can be recommended at this time, reducing the incidence of post prostatectomy urinary incontinence should be the goal. PMID- 9739279 TI - Behavioral treatment of urinary incontinence: a complementary approach. AB - When treating urinary incontinence, the first choice for treatment should be the least invasive and with the fewest side effects. Behavioral intervention is one such treatment modality. Behavioral interventions are defined as a change in the relationship between the patient's symptoms and his/her environment. Behavioral interventions include fluid and dietary management, toileting assistance, bladder retraining, urge suppression, and pelvic muscle rehabilitation. Intermittent self catheterization is also considered a behavioral intervention when combined with fluid/dietary or toileting changes. Before selecting a treatment option, a detailed history, physical examination, and environmental assessment should be taken. In addition, to effectively employ behavioral interventions, the patient's goals should be determined, the patient should be taught about the underlying causes of his/her problem, and he/she should be provided with not only motivation, but also support. Finally, behavioral interventions recognize the significant contribution that the patient makes in his/her own recovery. PMID- 9739280 TI - Pelvic floor rehabilitation: conservative treatment for incontinence. AB - Pelvic floor rehabilitation is used to treat stress urinary incontinence, urge, and fecal incontinence as well as other pelvic floor musculature disorders. When treating patients, it is important to thoroughly assess the pelvic floor. In addition to evaluating the urinary system, sexual and bowel functions must also be considered. Treatment plans should be devised on an individual basis according to the evaluation findings. Rehabilitation goals should be established. The patient must understand the function of her urinary system and the role she must play in its control. Muscle retraining is achieved through a personalized exercise program. This program may be augmented by manual techniques, biofeedback or electrical stimulation. While the Agency for Health Care Policy and Research (AHCPR) does endorse the use of behavioral modalities in treating urinary incontinence, the use of bladder retraining and pelvic floor rehabilitation is not always recommended when indicated, nor accessible for all patients who require it. More research is needed, in addition to ongoing public and professional education on behavioral interventions in order to underline the advantages of this form of treatment for incontinence. PMID- 9739282 TI - [ROL celebrated its 20 anniversary] [In Process Citation] PMID- 9739281 TI - [Quality has its price]. PMID- 9739283 TI - [Treatment of overweight. Use of a program]. AB - This article publicizes the results of an overweight treatment program carried out as a working model under the auspices of Primary Health Care. This program placed special emphasis on educational aspects, preventive as well as promoting healthy ways of living; and one in which activities planned towards establishing means of conduct which play a direct role in their users health factors are focused on. This treatment program occurred as a group activity. The 27 participating subjects, 23 women and 4 men, were recommended by their doctor/nurse. Their average age was 37.6 years, +/- 11.3. 19 subjects were overweight with an IMC between 25 and 30; the other 8 had been diagnosed as obese with an IMC over 30. This program took place in the Health Center meeting hall over 4 months, from February through June. Among the elements of this program, these deserve special mention: the use of a vocabulary exempt of adverse connotations, for example, instead of diet we spoke of menus; the establishment of behavioral goals before weight loss goals; the practice of elaborating and designing hypercaloric menus; providing positive reinforcement for those behaviors which increased the level of physical activity which was objectively measured by means of podometers; the practice of eating slowly; etc. At the end of the program, the results showed an average weight loss of 4.2 kilograms, a range of 0.9 to 6.12 K, with respect to the average weight at the start of the program. One can foresee that this loss will continue to occur, especially in those cases where the subject had modified some habits such as eating rapidly or performing physical exercise related to overweightness. These results are open to discussion regarding the efficiency of the program and with regards to this program as an example of a coordinated effort between the Mental Health Unit and a Primary Health Care Team in relation to the design and implementation of health programs directed towards the attention of people with overweight problems. PMID- 9739284 TI - [Nursing care for myelomeningocele in infancy]. AB - After studying the factors which most often lead to the hospitalization, recuperation and treatment facilitating release from hospitalization of newborns affected by myelomeningocele, we present this treatment plan. One commences with an evaluation, after which various problems are detected. Once identified, establish priorities setting objectives to meet and their corresponding dates; continue by drawing up an action plan which lists those activities that help achieve one's objectives. The purpose of this plan is to provide individualized quality treatments as well as to create a relaxed, confidential atmosphere in which parents express their fears while becoming secure in their practice of those treatments their child needs in their home. PMID- 9739285 TI - [Strategies of power and leadership for the development of nurses' social compromise]. AB - This article presents an analysis about power and leadership regarding the nursing profession in Spain and poses some strategies which, from the aforementioned perspective, allow the compromise nurses have with society to develop. This article has three sections: one which defines terms such as power, leadership, representative, corporation, ...; a second section which analyzes and makes proposals, wherein the current situation of the nursing profession is described in relation with the elements defined as the shapers of professional power, and proposals are posed along the lines of one to demand clear guidelines for nurses' professional role, to act in a "corporational" way, in positive terms, or to recognize the importance "informal" power networks hold; and finally, a section of conclusions. PMID- 9739286 TI - [A motivating experience: intrahospital teachers]. PMID- 9739287 TI - [The level of anxiety and depression in shut-in women]. PMID- 9739288 TI - [Nurses of one mind. Reasons for a conviction]. PMID- 9739290 TI - [Neck showing the carotid and subclavian arteries (superficial dissection on the right)]. PMID- 9739291 TI - [Orthopedic shoes and their modifications]. AB - The bone structure of the foot, knee and hip and their principle deformities are presented. With this in mind, the orthopedic shoe and its modifications are discussed as an extremely important element in correcting deformities that cause walking alterations, pain, etc.... How the shoes are made, the basic characteristics they should possess, which shoes are appropriate for which age, and problems the wrong shoe type can provoke are also discussed. PMID- 9739289 TI - [Carriers of hepatitis B and C virus, level of their information]. AB - OBJECTIVES: 1) To determine the degree of health information patients infected by Hepatitis B or C have regarding their illness. 2) To ascertain if relatives of these patients are also infected by Hepatitis B or C. METHOD: This was an observational cross study which analyzed the data obtained from 100 questionnaires filled out by patients infected by Hepatitis B or C under the control of the Digestive Pathologies Dispensary at the Sant Pau Hospital in Barcelona. The questionnaire was designed in order to evaluate the degree of information patients have concerning: A) means of transmission, B) preventative measures and their use, C) home/family environment. RESULTS: 85 patients indicate they had received information; of these, 73% consider their information to be satisfactory and of these, only 24% were familiar with the means of transmission this disease has. Finally, only 11 stated they correctly followed preventative measures. CONCLUSIONS: 1) The vast majority of the patients affected by Hepatitis B or C have a deficient level of information concerning the means of transmission and of prophylactic measures. 2) The prevalence of intrafamily infection cases studied is high. 3) We conclude that patients infected by Hepatitis B have significantly more information regarding the means of infection than do those patients infected by Hepatitis C. 4) The majority of the patients studied do not recognize the educational role nurses have concerning their health. PMID- 9739292 TI - [Nutrition related to HIV-AIDS infection]. AB - The author analyzes the importance of correct nutrition for all those patients affected by HIV-AIDS infection due to the fact that for quite a while a relationship has been shown to exist between the quality and quantity of digested nutrients and the nutritional state of a patient and his/her immunocompetence. The author reviews the possible nutritional state affections and the most appropriate diets during the various phases of this disease. A few specific recommendations are made and a few diets which increase calories or proteins are presented. PMID- 9739293 TI - Publicity. PMID- 9739294 TI - Creating a Coumadin Clinic. PMID- 9739295 TI - Magnet program recognizes excellence in nursing. PMID- 9739296 TI - Searching for an ideal blood substitute. PMID- 9739297 TI - Keeping pace with implanted defibrillators. PMID- 9739298 TI - New guidelines for breastfeeding. PMID- 9739299 TI - Fighting cancer fatigue. PMID- 9739300 TI - Look for signs of abuse. PMID- 9739301 TI - Elder abuse: what the law requires. PMID- 9739302 TI - A therapeutic bee sting? PMID- 9739303 TI - When the migraine rx isn't working. PMID- 9739304 TI - Get involved! PMID- 9739305 TI - [Multiple meanings of apprenticeship. To learn and to develop. Interview by Pierrette Lhez]. PMID- 9739306 TI - [Nursing practice and specific skills]. PMID- 9739307 TI - [Knowledge transmitted during the initial education]. PMID- 9739308 TI - [Grandmother's remedies. A teaching experience]. PMID- 9739309 TI - [Key to the integration of competences in public health. Collaboration stage Institute of nursing education]. PMID- 9739310 TI - [Nursing care in emergencies and catastrophies. From the individual concrete case to the collective concrete case]. PMID- 9739311 TI - [From the imaginary, from arithmetic and dosage calculations towards a teaching protocol for dosage calculations]. PMID- 9739312 TI - [Loss of sense, sense of loss. Attachment, grief and separation]. PMID- 9739313 TI - [The sense of care in French Polynesia]. PMID- 9739314 TI - [Education. Theoretical evaluation. Evaluation in other ways: strategy and stakes]. PMID- 9739315 TI - [Hospital training and motivation of students. Nurses, nurses' aides and management]. PMID- 9739317 TI - [Feeling more comfortable in the hospital]. PMID- 9739316 TI - [It is high time to beat pain. Interview by Anne Boiteux]. PMID- 9739318 TI - [The little girl who was afraid of holes]. PMID- 9739319 TI - [Music in the hospital. A means of development of expression]. PMID- 9739320 TI - [Music and pediatric psychiatry]. PMID- 9739322 TI - [The nurse in the new health care system of Tchad]. PMID- 9739321 TI - [Voice, sounds and music; allies for taking care]. PMID- 9739323 TI - [The digestive tract of the small child. Psychological and psychoanalytical aspects]. PMID- 9739324 TI - [The cocoon. A means to avoid tying down children]. PMID- 9739325 TI - [What is social pediatrics?]. PMID- 9739326 TI - Differentiation of infection from vaccination in foot-and-mouth disease by the detection of antibodies to the non-structural proteins 3D, 3AB and 3ABC in ELISA using antigens expressed in baculovirus. AB - The baculovirus expression system was found to be efficient at expressing the 3D, the 3AB and the 3ABC non-structural proteins (NSP) of foot-and-mouth disease virus (FMDV) as antigens recognised by immune sera in ELISA. ELISA's using 3D, 3AB and 3ABC detected antibodies from day 8 and 10 after experimental infection of susceptible cattle and sheep and cattle remained seropositive for more than 395 days. The ELISA's detected antibodies against any of the seven serotypes of FMDV. The 3D ELISA was specific and precise and as sensitive as established ELISA's which measure antibody to structural proteins. The assay may be used as a resource saving alternative to established ELISA's for the detection of antibodies against any of the seven serotypes. The 3AB and the 3ABC ELISA were also specific and precise. FMDV infected cattle could be differentiated from those that had been merely vaccinated as they gave a positive result in both the 3AB and the 3ABC ELISA's. Two cattle that had been both vaccinated and infected also gave positive results in both tests, suggesting that the 3AB and 3ABC ELISA's, but not the 3D ELISA might represent a reliable means of detecting infection in a vaccinated population. PMID- 9739328 TI - Genomic sequence of physalis mottle virus and its evolutionary relationship with other tymoviruses. AB - The genome of physalis mottle tymovirus (PhMV) is 6673 nucleotides long and is rich in cytosine residues (40.58%) like other tymoviruses. The organization of the genes is also similar to that of five other tymoviruses whose sequences are known. However, PhMV has the longest 3' noncoding region as well as the longest replicase (RP) ORF. The RP sequences are similar to those of other tymoviruses (48-60% identity) whereas the coat proteins (CP) and the overlapping proteins (OP) are conserved to a lesser extent (30-50% and 26-34% respectively). A tetra peptide "GILG" was found to be present in all the tymoviral OPs. The PhMV RP also possesses the methyl transferase, polymerase and the helicase motifs found in all the Sindbis-like super group of plant viruses. A phylogenetic analysis of the six tymoviral sequences revealed that they do not have a rigid hierarchical similarity relationship. PMID- 9739327 TI - A serological study of canine herpes virus-1 infection in the English dog population. AB - An epidemiological survey investigated the prevalence of canine herpes virus-1 antibodies in a population of 325 pet dogs in England. Sera were analysed for the presence of canine herpes virus-1 neutralising antibody by means of a serum neutralisation test and for virus-specific IgG and IgM by means of enzyme-linked immunosorbent assays. In contrast with published results from other parts of the world, canine herpes virus-1 infection was shown to be common among the domestic dog population of England. PMID- 9739329 TI - Avian polymavirus in wild birds: genome analysis of isolates from Falconiformes and Psittaciformes. AB - Avian polyomavirus (APV) infections have been reported to cause fatal disease in a wide range of psittacine species. Here we demonstrate APV infections in buzzards (Buteo buteo) and in a falcon (Falco tinnunculus) found dead in Germany, and in lovebirds (Agapornis pullaria) with fatal disease, wild-caught in Mocambique. APV infection in buzzards was determined by PCR amplification of parts of the viral genome followed by Southern blot hybridisation. The genomes of the isolates obtained from the falcon and one of the lovebirds proved to be very closely related to those of Budgerigar Fledgling Disease Virus (BFDV)-1, BFDV-2 and BFDV-3, isolated from budgerigar, chicken, and parakeet, respectively. A consensus sequence was delineated from the known nucleotide sequences of APV isolates. The significance of some nucleotide changes is discussed. Infectivity of all of these isolates was neutralized by antibodies directed against BFDV-1. Data presented in this investigation show that the polyomavirus isolates obtained from different avian species so far all belong to one genotype and one serotype within the proposed subgenus Avipolyomavirus of the family Papovaviridae. The designation Budgerigar Fledgling Disease Virus (BFDV) is, therefore, misleading as this virus type infects different species of birds. The name Avian Polymavirus and the abreviation APV should be adopted to all of the isolates investigated in detail at present. The possible role of birds of passage in the epidemiology in APV infections is discussed. PMID- 9739330 TI - Computer-aided virus identification on the World Wide Web. AB - An attempt has been made to devise computer software that will aid virologists to identify unknown virus isolates using the World Wide Web. Computerized information from the Animal Virus Information System was used to obtain data on various characters of a virus species. Sequence data banks are used to obtain the molecular data. A probabilistic method of virus identification based on Willcox's implementation of Bayes' theorem is implemented. The program provides hints to the users to carry out additional tests required to obtain higher confidence in identification of virus species. Signature peptides of the virus can also be used to confirm identification. The software is implemented on a UNIX machine and is written in C, UNIX shell scripts and HTML to run on the World Wide Web. This is the first species identification software that allows the user to carry out identification online through Internet. PMID- 9739331 TI - Haemagglutinin-esterase protein (HE) of murine corona virus: DVIM (diarrhea virus of infant mice). AB - The acetylesterase (AE) activity of DVIM (diarrhea virus of infant mice) was assigned to the haemagglutinin-esterase (HE) protein. The substrate specificity was examined using the natural substrate bovine submaxillary mucin (BSM) and/or synthetic substrates p-nitrophenylacetate (p-NiA) and alpha-naphthylacetate (alpha-NA) and compared with several strains of MHV and influenza viruses. The AE of DVIM hydrolyzed the O-acetylester bond of BSM, and the two synthetic substrates p-NiA and alpha-NA in vitro. MHV-S reacted efficiently with both p-NiA and alpha-NA but less with BSM. Influenza virus (C/Miyagi/77) reacted with BSM efficiently, however reacted with p-NiA weakly, but not with alpha-NA at all. Thus, the AE-reactivity of DVIM was distinctly different from that of MHV-S and influenza C virus, suggesting that the AE of HE may have a modified function. Isolation of HE by the treatment with non ionic detergent NP40, resulted in globules approximately 5 nm in diameter. DVIM-binding proteins were demonstrated in the plasma membrane of mouse intestinal brush-border cells and hepatocytes. The same protein was recognized by MHV-S and MHV-4. The cell membranes obtained from these target tissues were substrates for the AE of DVIM. The biological importance of the HE protein for DVIM is discussed. PMID- 9739332 TI - Nucleotide sequence of both genomic RNAs of a North American tobacco rattle virus isolate. AB - The complete sequence of a North American tobacco rattle virus (TRV) isolate, 'Oregon yellow' (ORY), was determined from cDNA and RT-PCR clones derived from the two genomic RNAs of this isolate. The RNA-1 is 6790 bases and RNA-2 is 3261 bases. The sequence of TRV-ORY RNA-1 was similar to RNA-1 to TRV isolate SYM, and differs in 48 nucleotides. TRV-ORY RNA-1 was one base shorter than--SYM, and had 47 base substitutions resulting in 12 amino acid substitutions of which 4 were conservative. The RNA-2 of TRV-ORY was distinct from RNA-2 of other characterized TRV isolates and contained three open reading frames (ORFs) that could potentially code for proteins of MW 22.4 kDa, 37.6 kDa and 17.9 kDa. Based on the homology of the predicted amino acid sequence with those of other tobraviruses. ORF1 of RNA-2 encodes the coat protein (CP). The protein sequence of ORF2 had regions of limited similarity with those of ORF2 of two other TRV isolates and pea early browning tobravirus. The ORF3 was unique to TRV-ORY. Phylogenetic analysis of tobravirus CPs indicated that TRV-ORY was most closely related to pepper ringspot tobravirus and TRV-TCM. The relationship of tobravirus CPs to other rod-shaped tubular plant viruses is also discussed. PMID- 9739333 TI - High-dose (9 MU) long-term (60 weeks) alfa-interferon therapy for chronic hepatitis patients infected with HCV genotype 1b. AB - Efficacy of standard regimens (e.g., 3-6 MU for 24 weeks) of alfa-IFN therapy for chronic hepatitis C has been limited, particularly in patients with HCV/1b. To see if higher-dose longer term treatment is more effective, we tried a 9 MU 60 week regimen. HCV/1b-infected chronic hepatitis patients received 9 MU IFN alpha 2a everyday but Sunday for 2 weeks and thrice a week for next 10 weeks, and 76 patients became HCV RNA-negative while 81 remained positive. The RNA-negative patients were then randomized to receive 3 MU (group I, n = 37) or 9 MU (group II, n = 39) for 48 weeks. Of the RNA-positive patients, only those with normal ALT received another 9 MU 48-week treatment (group III, n = 45). Sustained responders (SR) were defined as those with negative RNA and normal ALT 6 months after the therapy. SR rates based on intent-to-treat principle did not differ significantly between groups I and II (30% vs 41%), but those based on the protocol-compatible cases showed a significantly lower than those in group II. Adverse effects of IFN, developed more frequently in groups II and III than in group I, were mostly reversible. In conclusion, our results encourage 9 MU 60 week IFN alpha treatment in HCV/1b-infected patients with careful attention to adverse effects, and suggest that the treatment should be discontinued if HCV RNA does not disappear within 12 weeks. PMID- 9739334 TI - Effects of bafilomycin A1 on Japanese encephalitis virus in C6/36 mosquito cells. AB - Involvement of intracellular acidic compartments in the early phase of Japanese encephalitis (JE) virus infection of C6/36 mosquito cells was examined by bafilomycin A1, a specific inhibitor of vacuolar type H(+)-ATPase (V-ATPase). Dose dependent reduction of viral envelope protein (E) produced into the infected culture fluid was observed by pretreating the cells with 0.25 to 1.0 microM bafilomycin A1. In synchronized infection, cell surface-bound virions were internalized immediately by heating at 31 degrees C, followed by the release of nucleocapsid into the cytosol within a short lag period. Subcellular distribution of infecting 3H-uridine-labeled viral RNA (V-RNA) and its RNase sensitivity were analyzed by fractionation in Percoll density gradient centrifugation. At a 10 min chasing period, an RNase resistant V-RNA peak was found in fractions with a mean density of 1.05 g/ml corresponding to the endosome, while an RNase sensitive V RNA peak was detected at density range of 1.052-1.054 g/ml corresponding to the ribosome in C6/36 cell homogenate. The results indicate that JE virus infection in C6/36 cells proceeded through the endocytic pathway involving intracellular acidic compartments which was affected by bafilomycin A1. PMID- 9739335 TI - Evolutionary characteristics of influenza B virus since its first isolation in 1940: dynamic circulation of deletion and insertion mechanism. AB - New antigenic variants of B/Yamagata/16/88-like lineage which appeared in the season of 1997 as a minor strain tended to predominate in the following season. Also, we could observe for the first time, three peaks of activity caused by H3N2 virus and two variants of B influenza virus. Antigenic and phylogenetic analyses revealed that B/Victoria/2/87-like variants appeared again in Japan in 1997 after a nine-year absence. Influenza B viruses evolved into three major lineages, including the earliest strain (I), B/Yamagata/16/88-like variants (II), which comprised of three sublineages (II-(i), II-(ii), II-(iii)), and B/Victoria/2/87 like variants (III). Evolution of influenza B virus hemagglutinin was apparently distinguishable from that of influenza A virus, showing a systematic mechanism of nucleotide deletion and insertion. This phenomenon was observed to be closely related to evolutionary pathways of I, II-(i), II-(ii), II-(iii) and III lineages. It was noteworthy to reveal that the nucleotide deletion and insertion mechanism of influenza B virus completed one cycle over a fifty-year period, and that a three nucleotide deletion was again observed in 1997 strains belonging to lineage II-(iii). It was evident that amino acid substitutions accompanying nucleotide insertions were highly conserved. PMID- 9739337 TI - Virological and molecular parameters of HIV-1 infection of human embryonic astrocytes. AB - Two different strains of HIV-1, the lymphotropic HIV-IIIB and the monocytotropic HIV-Ba-L, were able to infect tertiary cultures of astrocytes established from the human embryonic brain. The infection did not require contact with infected cells, as astrocytes were exposed to infectious cell-free supernatants. Except for an early transient peak of p24 consistently observed after infection with HIV Ba-L, the infection of astrocytes appeared to be nonproductive. However, viral production was always observed when infected astrocytes were cocultured with permissive cells (CEM-SS or monocytes). To exclude the possibility that undetectable levels of virus are chronically produced by astrocytes, we exposed permissive cells to p24 negative supernatants taken from infected cultures. In such conditions permissive cells were never infected. Infection of astrocytes by HIV-1 was further supported by the finding that provirus persisted in these cells. Indeed, by a nested PCR, we detected HIV-1 DNA even one month after infection. Moreover, at the transcriptional level we observed expression of the multiply spliced RNA (tat and nef primers). Noteworthy, this pattern of HIV-1 expression did not change appreciably when astrocytes were pretreated and cultivated in the presence of IL-1 beta. Altogether, our data support the concept that astrocytes may play a role in the spread of HIV-1 infection within the brain and in the pathogenesis of neuro-AIDS. PMID- 9739336 TI - Phylogenetic analyses of the matrix and non-structural genes of equine influenza viruses. AB - Matrix (M) and nonstructural (NS) genes of thirteen equine H3N8 and H7N7 influenza viruses were sequenced and analyzed from an evolutionary point of view. The M and NS genes of H3N8 viruses isolated between 1989 and 1993 evolved into two minor branch clusters, including isolates from Europe and the American continent, respectively. It was noteworthy to reveal that the nucleotide sequences of the M and NS genes of an earlier American strain showed highest homology to those of recent European viruses. "Frozen evolution" was observed in the M and NS genes of A/eq/LaPlata/1/88. It was also evident that the NS gene of an H7N7 virus from 1977 was very similar to that of a 1979-H3N8 virus, while the M gene was closest phylogenetically to that of the earliest H7N7 virus isolated in 1956. Furthermore, the M2 protein of A/eq/Newmarket/1/77 virus contained a carboxyl terminal deletion of three amino acids. The evolutionary rates of the M and NS genes of H3N8 equine influenza viruses were estimated to be 5.4 x 10(-4) and 5.1 x 10(-4) substitutions per site per year, respectively, which were slower than those of human viruses. PMID- 9739338 TI - Synthesis of biologically active cDNA clones of cymbidium mosaic potexvirus using a population cloning strategy. AB - Biologically active cDNA clones of cymbidium mosaic potexvirus (CymMV) were synthesized using a population cloning strategy. Three populations of overlapping RT-PCR products encompassing the entire viral RNA of CymMV were ligated into pBluescriptKS+ with T7 RNA polymerase promoter fused to the 5' extreme of the viral cDNA. Capped-RNA in vitro transcripts were infectious. This is the first report of successful synthesis of biologically active CymMV clones. Unlike the conventional methods, population cloning maximizes the probability of obtaining biologically active cDNA clones. PMID- 9739339 TI - A temperate phage with cohesive ends induced by mitomycin C treatment of Lactobacillus casei. AB - A temperate phage, named PL-2, was induced from Lactobacillus casei ATCC 27092 by mitomycin C treatment of the cells at exponential growth phase. The phage had an isometric head of 45 nm in diameter and a flexible, non-contractile tail, 150 nm long and 10 nm wide, with a sharp tip. Along the tail axis, about 40 regularly spaced striae were seen. The phage DNA had complementary cohesive ends. The restriction enzyme map of the DNA was constructed by using 13 different restriction endonucleases. The size of the DNA was 35.2 kb, 83% in size of that of phage PL-1 lytic for the same Lb. casei strain. PMID- 9739340 TI - Critical amino acid changes in VP2 variable domain are associated with typical and atypical antigenicity in very virulent infectious bursal disease viruses. AB - Classical serotype 1 infectious bursal disease viruses (IBDV), but not very virulent (vv) isolates, react with neutralizing monoclonal antibody (NMab) 3 in virus neutralization tests or antigen-capture ELISA. Two other NMabs, 6 and 8, bind to both classical and most vv strains, but not to the atypical 94,432 and 91,168 vv strains, respectively. The basis for such reactivities was investigated by sequencing the genome region encoding the VP2 major immunogenic domain. In classical, variant, vaccine or vv IBDV strains, negative reactions with NMab3 were associated with changes in the Proline-Glycine pair at amino-acid (aa) positions 222-223 (hydrophilic peak A), and negative reactions with NMabs 6 and 8 with aa changes from positions 318 to 324 (hydrophilic peak B). The 91,168 and 94,432 viruses are the first vvIBDVs to present aa changes in peak B. PMID- 9739341 TI - Watermelon bud necrosis tospovirus is a distinct virus species belonging to serogroup IV. AB - The nucleocapsid protein gene of a tospovirus infecting watermelon in India was cloned and sequenced. Sequence analyses showed that the gene was most closely related to those of watermelon silver mottle tospovirus (WSMV) from Taiwan and peanut bud necrosis tospovirus (PBNV) from India, the two definitive species of serogroup IV. Amino acid sequence similarity was 84% and 82% with WSMV and PBNV, respectively. On the basis of the sequence divergence and the previously determined host range differences, the watermelon tospovirus, designated as watermelon bud necrosis tospovirus, should be considered as a distinct species belonging to serogroup IV. PMID- 9739342 TI - Cardiothoracic trauma. PMID- 9739343 TI - [Diastolic function parameters and atrial arrhythmias in patients with arterial hypertension]. AB - OBJECTIVE: To investigate in patients with arterial hypertension (HT) the extent of left ventricular (LV) hypertrophy and diastolic function in relation to atrial arrhythmias. PATIENTS AND METHODS: In 112 hypertensive patients (40 women, 72 men; mean age 50 +/- 6.6 years) with a mean systolic blood pressure for the cohort of 170 +/- 5 mmHg, their first invasive coronary angiography was performed between July 1995 and October 1997 because of angina pectoris and/or an abnormal stress electrocardiogram. After excluding coronary heart disease LV dimensions and diastolic function were measured by echocardiography; in 59 of the 112 patients LV hypertrophy was demonstrated. In addition, long-term blood pressure monitoring, exercise and long-term electrocardiography, late-potential analysis and measurement of heart rate variability were undertaken. The control group consisted of 51 patients without arterial hypertension after exclusion of coronary heart disease. RESULTS: Even in the hypertensive patients without LV hypertrophy diastolic LV function and ergometric exercise capacity were reduced. The risk of LV arrhythmias was significantly higher in patients with LV hypertrophy than those without and in the control group, as measured by the complexity of atrial arrhythmias (P < 0.001), the incidence of abnormal late potentials (P < 0.001) and reduction in heart rate variability (29.3 +/- 5.3 ms vs 47.8 +/- 12.1 ms vs 60.7 +/- 6.6 ms; P < 0.001). There were similar results regarding severe complex atrial arrhythmias (38.5 vs 15.0 vs 0%; P < 0.001). The incidence of atrial arrhythmias correlated with the LV diameter (r = 0.68, P < 0.001), LV morphological dimensions and diastolic function (isovolumetric relaxation time r = 0.44, P < 0.001) and the ratio of early to late diastolic inflow (r = 0.46; P < 0.001). CONCLUSIONS: Hypertensive patients have a higher risk of atrial and ventricular arrhythmias, depending on the degree of LV hypertrophy. But atrial arrhythmias, in contrary to ventricular arrhythmias, are also closely related to abnormalities in LV diastolic function. PMID- 9739344 TI - [Full working capacity instead of threatened retirement. Thrombendarterectomy in chronic thromboembolic pulmonary hypertension]. AB - HISTORY AND CLINICAL FINDINGS: A 56-year-old man in marked right heart failure (stage III-IV of the New York Heart Association classification) and severe pulmonary hypertension was admitted to a rehabilitation clinic for therapeutic and social-medical assessment. On physical examination the important features were markedly distended neck veins, tachycardia at rest (90/min), a loud 2nd pulmonary sound and dyspnea. INVESTIGATIONS AND DIAGNOSIS: Non-invasive tests (ECG, echocardiography, abdominal ultrasound and lung functions) confirmed right heart failure; invasively obtained haemodynamic data indicated its severity. Selective pulmonary angiography defined the embolisation to be central and bilateral. TREATMENT AND COURSE: As intensive drug treatment and physiotherapy had failed to achieve significant improvement, operative removal of the bilateral central and some segmental pulmonary thrombi was performed and an inferior vena caval filter inserted. Immediately after operation the markedly elevated right heart and pulmonary artery pressures fell markedly and there was dramatic improvement in the patient's general condition and in his physical capacity. Angiography demonstrated largely normal pulmonary perfusion. Instead of the anticipated retirement, the patient was discharged on anticoagulants, in the expectation of a return to full-time work. CONCLUSION: With pulmonary thrombendarterectomy severe chronic thromboembolic pulmonary hypertension may well be treated. PMID- 9739346 TI - [CADASIL: clinical aspects and diagnosis]. PMID- 9739345 TI - [A case of Carney complex]. AB - HISTORY AND CLINICAL FINDINGS: For some months a 57-year-old woman had noted increasing shortness of breath, associated in the last few weeks with undirected vertigo and several brief periods of lost consciousness. She was finally admitted because of additional central facial paresis. On auscultation a high-frequency systolic murmur was heard over the apex and a discrete diastolic murmur over Erb's point. There were numerous facial freckles and three cutaneous myxomas. INVESTIGATIONS: Echocardiography revealed irregular tumours throughout the left atrium and a large broad-based one prolapsing through the mitrale valve in diastole. Computed tomography demonstrated a 6 x 6 cm tumour in the left lower abdomen, probably arising from the left ovary, and a second 3 x 3 cm presacral tumour. TREATMENT AND COURSE: At cardiac surgery four tumours were found in the left atrium and resected: histologically they were benign myxomas. Removal required extensive resection in the area of the interatrial septum and the atrial wall, resulting in 2 degrees AV block for which a VDD pacemaker was implanted. CONCLUSION: Atrial myxomas may be the cardinal sign of the Carney Complex, an autosomal dominant syndrome with cutaneous myxomas, myxoid abdominal tumours, hormone-producing tumours in the testicles, adrenal cortex or hypophysis, schwannoma as well as lentigines. For this reason, further tumours should be looked for if freckles and/or cutaneous tumours are found in association with an atrial myxoma. The patient and family should be informed about the genetic aspects. PMID- 9739347 TI - [Pericardial drainage: practical aspects]. PMID- 9739348 TI - [Hepatitis A prevention by vaccination]. PMID- 9739349 TI - [Volume manometry for determination of rectal compliance]. PMID- 9739350 TI - [The significance of the sympathetic nervous system during therapy for hypertension and related pathologies. Imidazoline-I1-receptor agonists. 17th Scientific Meeting of the International Society of Hypertension. Amsterdam, June 7, 1998]. PMID- 9739351 TI - [Why regular physical activity favors longevity]. AB - Regular physical exercise is useful at all ages. In the elderly, even a gentle exercise programme consisting of walking, bicycling, playing golf if performed constantly increases longevity by preventing the onset of the main diseases or alleviating the handicaps they may have caused. Cardiovascular diseases, which represent the main cause of death in the elderly, and osteoporosis, a disabling disease potentially capable of shortening life expectancy, benefit from physical exercise which if performed regularly well before the start of old age may help to prevent them. Over the past few years there has been growing evidence of the concrete protection offered against neoplasia and even the ageing process itself. PMID- 9739352 TI - [Correlation between functional and radiographic data, and broncho-alveolar cytology in patients with diffuse interstitial lung disease]. AB - BACKGROUND: "Interstitial Lung Disease" is a term that includes a heterogeneous group of disorders characterized by variable degrees of parenchymal inflammation (alveolitis) and fibrosis. Alveolitis represents one of the most important aspects of ILD and its characterization by broncho-alveolar lavage (BAL) can give explanations of the different clinical features, in association with the evaluation of physiological changes and High Resolution Computed Tomography (HRCT). The aim of this study was to establish a correlation between variations in broncho-alveolar cells and anatomical-functional alterations in patients affected by ILD before and after corticoid therapy. METHODS: Thirty-one patients (14 males 17 females) were introduced in the study and were evaluated for clinical signs, functional parameters, HRCT and cyto-immunological aspects of BAL fluid. All data were acquired at the admission in the study, after 60 and 180 days. Patients had received prednisone 1 mg/kg/die for 60 days and, then, 0.25 mg/kg/die for 120 days. RESULTS: In lymphocyte alveolitis, a clinical radiological improvement was frequently observed after therapy while in those with neutrophilic and eosinophilic components, no improvement was detected. CONCLUSIONS: In conclusion, BAL represents a useful tool in qualifying ILD which can be easily monitored by clinical, functional and radiological data. Therefore, a new method for the diagnosis and monitoring of ILD is suggested. This method has the marked advantage to reach a reliable diagnosis even in a high percentage of patients who refuse or can not undergo the biopsy assay. PMID- 9739353 TI - [Epidemiological and clinical findings in 697 syncope events]. AB - BACKGROUND AND AIMS: The PS structures comprise an epidemiological observatory of syncopal episodes (SE). Only a few studies of this sort are available in Italy: the data reported here represents an epidemiological and clinical contribution to the evaluation of this pathology. METHODS: The retrospective study was based on PS registers and on medical records, enrolling all patients aged > 13 years old who attended the PS between 1/1/1992 and 31/12/1992 for syncopal or near syncopal episodes. RESULTS: During the period in question, a total of 697 SE were observed in 684 patients (363 males and 321 females) with an incidence of 268 x 10(5)/year (1.3% of all admissions to PS); 530 SE (76%) were admitted to hospital (3.6% of all emergencies), 78.1% of which to the Emergency Medical ward. Intrahospital mortality was 13 SE. The following parameters were assessed: mean age 57.5 years (range 15-93), distribution by age bracket, place where SE occurred, posture and any trauma, present in 197 SE (28.2%), including 27 cases of bone fracture; 180 SE (25.9%) reported at least one similar earlier episode and 13 patients showed recurrence during the study period; the main potentially correlated chronic pathologies (the most frequent were arterial hypertension, CIC, diabetes mellitus), pharmacological treatment; the diagnostic assessment on discharge for the entire series (cardiogenic 12.6%, non-cardiogenic 69.6%, unknown genesis 16.9%) and in the case of hospitalised SE alone, the concordance was evaluated between diagnosis at admission and discharge (positive in 60.9%); the use of instrumental diagnostics and their diagnostic contribution. CONCLUSIONS: The incidence of syncopal pathology was confirmed to be high in PS, more so in patients admitted to hospital. Compared to other series, there were more SE caused by vasovagal mechanisms, ortyhostatic hypotension and psychogenic causes which together accounted for 46.3%, those caused by cerebral vasculopathies and supraventricular hyperkinetic arrhythmia at onset. PMID- 9739354 TI - [Genes, molecules, and mechanisms regulating programmed cell death]. AB - Apoptosis is a morphologically distinct form of programmed cellular death that plays a central role during embryogenesis, tissue homeostasis, and to remove not necessary or potentially dangerous cells. Moreover, disregulation of genes mediating or modulating apoptosis contributes to the pathogenesis of a number of human diseases, including cancer, autoimmune diseases, neurodegenerative disorders, viral infections and acquired immunodeficiency syndrome. A number of genes and molecules promoting or protective against cell death is at present-day known and an important information about the external and internal signals involved in stimulation and suppression of apoptosis is also emerging. In the intracellular pathway of the death deregulation of [Ca2+](i) plays a pivotal role. Increased ionized intracellular calcium stimulates both the activation of enzymes (protein kinases, endonucleases, proteases and phospholipases) and plasma membrane K+ channels. This calcium-mediated activation leads to morphostructural changes, such as cell shrinkage, cytoplasmatic blebbing, nuclear chromatin condensation and DNA degradation into oligonucleosomal fragments. At least some genes of the cell death pathway have been conserved throughout animal evolution; ced-3 e ced-9 that regulate the initiation of cellular suicide in the nematode Caenorhabditis elegans are homologous to genes that in mammalian cells are thought to play a similar role (interleukin-1 beta converting enzyme [ICE] family, Bcl-2). It is possible to suppose that these regulators could constitute a target for treatment of disorders related with disregulation of apoptosis. PMID- 9739355 TI - [Cellular adaptation and cancerogenesis]. AB - The paper describes the main adaptive mechanisms involved in the carcinogenic process. As a result of the action of carcinogenic agents (physical, chemical, biological), and in relation to the functional status of the affected cells, a number of systems are triggered off: detoxification and conjugation systems, the metabolisation of the said agents, DNA repairing enzymes, increased shock proteins (HSP), the induction of clonal proliferation. All these systems are valuable to the survival of the body and the species and culminate in the apoptosis of damaged cells as the last attempt at adaptation of a social kind for the good of the body. When these compensation mechanisms prove ineffective, imprecise or are exceeded by cell adaptive capacity, the resulting structural and functional alterations trigger off (induction) a very long process which often lasts between one and two thirds of the body's life, in various stages, multistep and multifactorial: this neoplastic transformation leads to a purposeless, egoistic, anarchic proliferation of cells which wish to survive at all costs, even to the detriment of the body of which they form part. Following the exhaustion of cell adaptive defences, there is an accumulation of additional genetic alterations (promotion and progression), the cells become manifestly neoplastic and continue their egoistic adaptation, according to the laws of natural selection: the cells which survive are those which adapt best to the hostile environment of the host's body, which are unaffected by proliferation control mechanisms (contact inhibition, differentiation factors, apoptosis, etc.), which make the best of the growth factors present in their microenvironment, which accomplish the so-called decathlon of the metastatization process, namely acquiring new capacities which can overcome the basal membrane, invade tissues to which they are attracted and continue to proliferate. Manifestly neoplastic cells become not self at a later stage, managing to escape the immune system using various adaptive mechanisms which induce immune tolerance/anergy. From this point of view, cancer may be regarded as an incidental factor in the host's cell adaptation processes; the latter are much more important from a biological point of view and their absence is incompatible with life: cancer might therefore be regarded as a cell adaptation pathology. PMID- 9739356 TI - [Role of the oncologist in prevention, diagnosis and treatment of neoplasms]. AB - Oncological pathology represents a topic of particular interest and importance owing to both its rising frequency (151,690 deaths in Italy in 1992) and the continuous progress made in the diagnostic and therapeutic field. The steady growth of this pathology has led to the need to identify a new professional figure: the cancer physician capable of bringing together all the skills required to optimise the diagnosis and treatment of oncological patients. As it is now universally accepted in theory, but not always put into practice, the treatment of oncological pathologies calls for a multidisciplinary approach. The optimal treatment is based on the integration of the roles of all the various specialists: oncologist, surgeon, radiotherapist and expert in pain therapy. In this therapeutic strategy, the cancer physician should manage the starting phase of the disease and subsequently should manage its course together with the surgeon and radiotherapist, returning in the terminal phase to sole management of the cancer patient with the help of other qualified professional figures. In short, the authors feel that it is not exaggerated to state that the cancer physician plays a central role in resolving the needs posed by the problem of cancer, needs which may take the form of diagnosis, therapy and care. PMID- 9739357 TI - [The filmless hospital--a legal challenge]. AB - PURPOSE: This article concerns the legal aspects of digital archiving of radiological images, in particular with regard to the resolution passed by the 37th Committee meeting of the Federal Board of "RoV" (x-ray ordinance). Concepts of filmless hospitals must therefore be tested for compliance with all present legal requirements. MATERIAL AND METHODS: The literature concerning the legal aspects of digital medical archiving of x-ray documents is reviewed. The concept of a digital archive was compared with these aspects and tested for compliance with the respective legal rules. RESULTS: There are no legal obstacles to realizing our concept of a filmless hospital. However, there are no civil court rulings on record, since there have not been any legal proceedings to date. CONCLUSIONS: There are no legal objections to a complete digital archiving in a hospital in the absence of any court rulings to the contrary. PMID- 9739358 TI - [Osteomyelitis--imaging methods and their value]. AB - Various imaging modalities are used in diagnosis of acute and chronic infectious endogenous osteomyelitis and exogenous ostitis. The pathophysiological changes of osteomyelitis/ostitis in the bone and surrounding soft tissue are known. Findings in plain film radiography show these changes only in relatively advanced stages of disease. Hence, plain film radiographs are useful as a basic imaging modality by excluding other differentials and as a follow-up modality under therapy. Ultrasound-using advanced technology--offers diagnostic help in acute osteomyelitis, especially in infants. The various techniques of nuclear medicine show much higher sensitivity for detecting osteomyelitis than plain film radiography, but do not permit good separation for bone involvement and infectious changes in the surrounding soft tissue. While computed tomography offers the ability to display bone and soft tissue separately, it has been widely replaced by magnetic resonance imaging using fat-suppressed sequences and paramagnetic contrast media which show the spread of the infectious changes with higher sensitivity and accuracy. PMID- 9739359 TI - [Postoperative imaging of synthetic coronary artery bypass graft patency by means of CT angiography]. AB - PURPOSE: CT angiography was performed in 12 patients with insufficient autologous graft situations to evaluate postoperative patency and situation of the proximal, distal and coronary artery anastomoses of synthetic Perma-Flow coronary artery bypass grafts. METHODS: Bypass grafts were evaluated postoperatively with spiral CT of the mediastinum. At a flow rate of 3 ml/s, 120 ml of contrast material were applied over a cubital vein. Slice thickness was 3 mm, maximum pitch factor 2 and image reconstruction was performed at 2 mm increment. Shaded surface displays were analysed together with axial scans for bypass evaluation. RESULTS: 8 out of 12 synthetic bypasses proved to be patent. One bypass was completely occluded and in three patients the distal portions of the grafts were occluded. Coronary angiography performed in one case confirmed complete bypass occlusion. Due to the occlusions, 8 distal and 12 proximal anastomoses were visible. Only 8 out of 19 side-to-side coronary artery anastomoses could be sufficiently well imaged with this technique. CONCLUSIONS: CT angiography is suitable for postoperative screening of synthetic coronary bypasses to determine the patency and anastomotic situations. Coronary artery anastomoses however are not sufficiently imaged and coronary angiography continues to be required. PMID- 9739360 TI - [Morphology and staging of primary mucosa-associated gastric lymphoma in hydro-CT imaging]. AB - PURPOSE: Evaluation by hydro-CT in diagnosing and staging of primary non-Hodgkin lymphoma of the stomach (MALT). MATERIAL AND METHODS: 15 patients with MALT lymphoma underwent imaging by hydro-CT (helical CT scanning optimised for parenchymal and vessel contrast with distension of the gastric wall by water). The CT scans were evaluated for the site, morphology, extent and contrast enhancement of gastric lymphoma; in addition, the number and location of abdominal lymph nodes were examined. The results of CT imaging were compared with the findings at endoscopy + biopsy and endosonography and in case of gastrectomy also with the histopathological results. RESULTS: All lymphomas were correctly diagnosed and were mostly located in the distal parts of the stomach. MALT lymphoma typically grew submucosally, infiltration of the mucosa was rare. Most tumours showed marked contrast enhancement of the mucosa and poor enhancement of the submucosa. Hydro-CT and endosonography had similar accuracies in respect of staging of compartment I and II lymph nodes. Staging of distant nodal groups was more accurate by hydro-CT. CONCLUSION: Hydro-CT is non-invasive and may be used for diagnosis and staging of primary gastric lymphoma with a typical morphology of gastric lymphoma. Hydro-CT may be regarded as complementary to endosonography and is well suited for the initial diagnosis of gastric lymphoma as well as for the diagnosis of recurrent tumour. PMID- 9739361 TI - [Magnetic resonance imaging (MRI) of liver and brain in hematologic-oncologic patients with fever of unknown origin]. AB - PURPOSE: To examine the advantage of liver and brain MRI in clinically anomalous haematological patients with fever of unknown origin. MATERIAL AND METHODS: Twenty liver MRI (T2-TSE, T2-HASTE, T1-FLASH +/- Gd dynamic) and 16 brain MRI (T2 TSE, FLAIR, T1-TSE +/- Gd) were performed searching for a focus of fever with a suspected organ system. Comparison with clinical follow-up. RESULTS: A focus was detected in 11/20 liver MRI. Candidiasis (n = 3), mycobacteriosis (n = 2), relapse of haematological disease (n = 3), graft versus host disease (n = 1), non clarified (n02). The remaining 9 cases with normal MRI were not suspicious of infectious hepatic disease during follow-up. In brain MRI, 3/16 showed a focus (toxoplasmosis, aspergillosis, mastoiditis). Clinical indication for an infectious involvement of the brain was found in 4/16 cases 2-5 months after initially normal brain MRI. No suspicion of an infectious involvement of brain was present in the remaining 9/16 cases. CONCLUSION: In case of fever of unknown origin and suspicion of liver involvement, MRI of the liver should be performed due to data given in literature and its sensitivity of 100%. Because of the delayed detectability of cerebral manifestations, in cases of persisting suspicion even a previously normal MRI of the brain should be repeated. PMID- 9739362 TI - [First clinical results of ultrafast, contrast-enhanced 2-phase 3D-angiography of the abdomen]. AB - PURPOSE: To assess the utility of breath-hold abdominal ultrafast three dimensional (3D) gadolinium-enhanced dual-phase magnetic resonance angiography (MRA). MATERIAL AND METHODS: 125 patients with various abdominal pathologies were imaged using a breath-hold ultrafast gadolinium-enhanced dual-phase 3D-MRA technique. RESULTS: 119 (95%) of 125 MRA's were of good or excellent quality. The sensitivity in the detection of renal artery stenoses as well as stenoses of the celiac trunk and the superior mesenteric artery was 100%. Accessory renal arteries (n = 9) and replaced hepatic arteries (n = 4) were reliably detected by MRA. In 24 (71%) of 34 cases MR-angiographic delineation of the spleno-portal system and hepatic veins was superior compared to conventional angiography. CONCLUSION: Breath-hold gadolinium-enhanced dual-phase 3D-MRA has the potential to replace conventional angiography in the abdomen. PMID- 9739363 TI - [Intraoperative findings and postoperative computer tomographic follow up of inflammatory aortic aneurysm]. AB - PURPOSE: Retrospective evaluation of postoperative long-term results after surgery of inflammatory aortic aneurysms (IAAA) with computed tomography (CT). Findings in CT were analysed with particular attention to the development of inflammatory tissue adjacent to the aneurysm site. MATERIAL AND METHODS: Of 2101 patients operated on an aortic aneurysm 5.4% (114 patients) presented typical intraoperative features of inflammatory aortic aneurysms. 54 of these 114 patients (47%) were examined via computed tomography pre- and post-operatively. On an average the follow-up-study was performed 2.5 years postoperatively. RESULTS: All follow-up-studies revealed a correct location of the aortic prostheses. In 85.1% of the cases there was either no or negligible persisting inflammatory tissue with a diameter of less than 2 mm. 10.6% of the patients demonstrated remaining but reduced inflammatory tissue. In 4.3% of the cases the extent of the inflammatory tissue had not changed. Aneurysms of the anastomoses (n = 4), morphologic renal changes (n = 7) and an aorto-enteric fistula were demonstrated by CT as postoperative complications. CONCLUSIONS: In evaluating recurrence of the aneurysm and possible complications as well as the development of the inflammatory tissue, postoperatively performed computed tomography proved a reliable diagnostic method. PMID- 9739364 TI - [Measurement of migration of acetabular components in cementless hip replacement]. AB - PURPOSE: Migration measurements of acetabular components using a special computer aided method (EBRA = abbreviation for the German term "Ein-Bild-Rontgenanalyse") were performed to evaluate early results of the implants and predict aseptic loosening. METHODS: Standard ap-radiographs of the pelvis were marked, specific points were digitised. Simulating the spatial situation the programme computes longitudinal and vertical migration of the cup. 74 acetabular components in 71 patients could be studied by migration measurements. RESULTS: 14 patients showed migration of more than 1 mm, which is the confidence limit of this method. Each of these patients showed diverse reasons for the migration, i.e. osteoporosis of the acetabular bone stock or problems concerning the surgical technique which means malposition of the cup or insufficient reaming of the bone. There were some patients with severe congenital dysplasia of the hip and in some cases the inclination angle of the cup was too great. CONCLUSION: The technique applied for measuring migration of acetabular components can be useful for evaluating early instability of the implant and can be helpful in detecting problems concerning the surgical technique. PMID- 9739365 TI - [Ultrasonography as a diagnostic measure in the rupture of fibular ligaments. Comparative study: sonography versus radiological investigations]. AB - PURPOSE: The present prospective clinical study was designed to verify the value of ultrasound examination concerning injuries of the fibular ligaments of the ankle using functional examinations under ultrasound supervision. METHOD: 34 patients who had sustained a lesion of the fibular ligaments of the ankle were compared with 42 healthy persons. The lateral malleolus and the collum tali were used as reproducible bony landmarks in ultrasound. The distance between those two points is first of all was measured in neutral position and later on under the influence of supinating force of 15 kp using a Telos fixation device. RESULTS: The average lateral opening of the upper ankle joint measured with the dynamic ultrasound examination was 4.4 mm (1-12 mm) in the group of patients who had been suffering from rupture of the fibular ligaments. In comparison the average lateral opening of the healthy contralateral ankle was 3.9 mm (1-9 mm). The control group showed an average lateral opening of 0.9 mm (0-5 mm). CONCLUSION: Direct ultrasonic visualisation of the fibular ligament is not yet reliable. The described method allows an estimation of the stability of the fibular ligaments even in cases of chronic instability. The ultrasound investigation method discussed is difficult to handle and for that reason it cannot replace the standardised x-ray examination. PMID- 9739366 TI - [MRI of knee ligaments: error analysis with reference to meniscus and anterior cruciate ligaments in an arthroscopic controlled patient cohort]. AB - PURPOSE: To categorise discrepancies in findings of the menisci and anterior cruciate ligament (ACL) between arthroscopy and MRI. MATERIALS AND METHODS: The MRIs of 236 patients were retrospectively analysed by an experienced radiologist without knowledge of clinical and/or operative findings. Discrepancies in arthroscopic findings were reevaluated together with the arthroscopist to determine their cause of error. RESULTS: The diagnostic accuracies for injuries of the medial and lateral meniscus and the ACL were 92.4%, 92.4%, and 94.1%, respectively. For the menisci, causes for discrepancies in findings (n = 31) were: overinterpretation of central signal intensities with contact to the meniscal surface but without disturbance of the meniscal contour as a tear (n = 12), insufficient arthroscopie evaluation of the knee joint (n = 11), overlooked tears on MR imaging (n = 6), misinterpretation of normal anatomic structures (n = 1), "magic angle" phenomenon (n = 1), and missed tears at MRI (n = 1). Causes for discrepancies for the ACL (n = 18) were: nearly complete versus complete rupture either at MRI or arthroscopy and vice versa (n = 9), insufficient arthroscopic evaluation (n = 6), insufficient MRI technique (n = 2), and overlooked tear on MR imaging (n = 1). CONCLUSIONS: Discrepant findings between MRI and arthroscopy may be also due to an insufficient arthroscopic evaluation in clinical routine. The close cooperation between surgeons and radiologists improves the understanding of the methods of each other. PMID- 9739367 TI - [Blood flow quantification in hemodialysis shunts by phase contrast magnetic resonance angiography (PC-MRA) compared with duplex sonography]. AB - PURPOSE: Aim of this study was to evaluate whether phase contrast MR angiography (PC-MRA) could provide additional functional information besides morphology in the assessment of haemodialysis fistulae. MATERIAL AND METHOD: Twenty-two patients (11 male, 11 female), aged 22-77 years, were examined. MR images were obtained with a 1.5 T Gyroscan ACS-NT (Philips, Best, Netherlands) using a high resolution wrap-around coil. In addition to MRA blood-flow measurements were performed with a gradient-echo sequence (TR 14 ms, TE 5-5.5 ms, flip-angle 15 degrees, 6 mm slice thickness, retrospective gating, matrix 96:128) in the venous and arterial section of the fistulae. Doppler flow measurements were performed at the same position with a Sonoline Elegra (Siemens AG, Erlangen) using a 7.5 MHz transducer. RESULTS: Both methods of flow-volume measurements showed a good correlation (r = 0.94 in the arterial section, r = 0.90 in the venous section, p < 0.001). The average calculated blood flow was measured 11% (arterial section) and 12.8% (venous section) higher with Pulsed Waved Doppler as compared to PC MRA. CONCLUSION: PC-MRA with a high-resolution wrap-around coil is a reliable method for measuring functional parameters like flow-volume and flow velocity in haemodialysis fistulae and a useful complement to the visualisation capabilities of MRA. PMID- 9739368 TI - [Dissections of the basilar artery]. AB - PURPOSE: To describe clinical, radiological and pathological-anatomical findings of basilar artery dissections. METHODS: During a period of three years (1994 1996) we observed 4 patients with dissections of the basilar artery, proven by angiography. Angiograms, MRI (n = 2) and autopsy results (n = 1) of these patients were correlated with clinical symptoms and long-term follow-up. RESULTS: In three patients the dissection was confined into the basilar artery. Three patients with vertebrobasilar ischaemia showed irregularities of the vessel wall on angiography, in one of these patients autopsy revealed a haematoma within the vessel wall, located between intima and media. One patient complaining of relapsing headaches had a posttraumatic basilar artery aneurysm. CONCLUSION: Dissections of the basilar artery can be separated in two types. In case of subintimal dissection patients present with vertebrobasilar ischaemia, which in contrast to extracranial dissections usually occurs without temporal delay. A second type of patients presents with subarachnoid haemorrhage. The dissection is subadventitial and pierces through a thin adventitia into the subarachnoid space. PMID- 9739369 TI - [CT angiography to determine the size of intracranial aneurysms before GDC therapy]. AB - OBJECTIVE: To examine in a model and in clinical practice whether CT angiography (CTA) is suitable to determine the size of intracranial aneurysms. METHODS: For an aneurysm model the contrast medium-filled balloon of an occlusion catheter was used. At different levels of filling images were obtained by both CTA and digital subtraction angiography (DSA). In the CT image the size of the simulated aneurysm was calculated from the CTA data at the workstation while from the DSA images it was performed by the DSA system with the use of both an external reference structure (two-ring method) and a stereotactic system (ANGIOLOG). In 7 patients, the size of the aneurysm was determined by CTA and DSA prior to aneurysm occlusion by means of guglielmi detachable coils (GDC therapy). RESULTS: On the basis of 2 D reconstructions of the CT average size deviations in the XY plane of 1.6% and on the Z axis of 3.2% were determined. These errors increased to 3.4% (XY plane) and 5.6% (Z axis) with 3 D reconstructions. By use of the two-ring model, DSA gave an average size deviation of 3% while with the stereotactic system (ANGIOLOG) it was as high as 11%. In 6 of the 7 patients, the appropriate spiral size was chosen primarily after CTA measurements. CONCLUSIONS: CTA enables the reliable determination of the size of an intracranial arterial aneurysm and in individual cases can give a better representation of an anatomic situation at the base of an aneurysm than DSA. Thus, CTA imaging of an aneurysm prior to GDC therapy is useful. PMID- 9739370 TI - [Verification of MR thermometry by means of an in vivo intralesional, fluoroptic temperature measurement for laser-induced thermotherapy ov liver metastases]. AB - PURPOSE: To evaluate the correlation of MR-measured changes of signal intensity and invasive fluoroptic temperature measurements during MR-guided LITT of liver metastases. MATERIALS AND METHODS: In 15 patients with proven liver metastases of colorectal carcinoma, MR-guided LITT was performed with a percutaneous approach in a multiapplicator technique. Two temperature sensitive T1-weighted sequences (FLASH-2D- and TurboFLASH-sequences) were used to map the spatial and temporal distribution of Nd:YAG laser effects. Parallel fluoroptic temperature measurements were carried out by means of an inserted probe in a distance of 5-26 mm (mean: 14 mm) from the laser applicator. RESULTS: In both sequences a gradually increasing signal loss could be documented during laser application which proved to be reversible after cessation of energy deposition. the percentage of decrease in signal intensity correlated directly with the measured increase of temperature. Invasive fluoroptical evaluation of temperature distribution after 10 min exposure time showed at 5 mm distance from the applicator an increase of temperature of 35 degrees C, in 10 mm distance a mean increase of 9 degrees C +/- 1.7, in 15 mm a mean increase of 7 degrees C +/- 1.6 and in 20 mm a mean increase of 3 degrees C +/- 0.5. This is evidence of thermal tissue damage up to 3 cm in diameter with laser monoapplication. The qualitative evaluation revealed a reproducible correlation of the extent of signal loss around the applicator and the finally induced degree of necrosis. CONCLUSION: Invasive fluoroptical temperature measurements prove the diagnostic reliability of MR thermometry for the online monitoring of LITT of liver metastases. PMID- 9739371 TI - [Radiologic implantation of central venous portal systems in the forearm]. AB - PURPOSE: To assess the safety and effectiveness of fluoroscopic guided brachial implanted central-venous miniport systems. PATIENTS AND METHODS: In 32 oncological patients a central-venous miniport system (Vital-Port, CPC-Cook) was implanted in the forearm. The group included 15 women and 17 men (range 33-78, mean 56 years). RESULTS: Technical success was 100%, in 6 patients vasospasm produced difficulties in catheter placement. In a total of 2878 patients days (range 3-445, mean 90 days) 5 complications occurred (15.6%, 1.7 on 1,000 catheterdays): In one patient (3.13%, 0.35/ 1000 d) the system was changed due to leakage of the catheter near the chamber. Four patients had minor complications: in one case excision of a suture line granuloma was necessary, one haematoma at the site of the chamber was treated with dressing, one patient had a wound dehiscence and one occluded catheter returned to patency by flushing the system with contrast material. Venous thrombosis, phlebitis, catheter dislocation, paravasation or system-related infection did not occur. CONCLUSION: Fluoroscopic guided peripheral central-venous port implantation is a safe and easy procedure with a high success rate and a low complication rate which can be performed in an outpatient setting. PMID- 9739373 TI - [Pitfalls when using a contrast media injector in MRI]. AB - PURPOSE: Using a power injector to applicate Gd-DTPA we found a contrast enhancement of the pyelo-caliceal system even in the native studies, and hence we analysed pitfalls when using power injectors in MRI. MATERIAL AND METHODS: We used a power injector Spectris (MedRad, Maastricht, Netherlands). In vitro artifacts were achieved by the mixture of contrast media and saline solution. We substituted contrast media by red water, NaCl by clear water. RESULTS: Using power injectors in MRI, some pitfalls must be avoided, which can render investigations useless, especially dynamic contrast-enhanced investigations. CONCLUSION: In our study we showed an easy way to overcome some pitfalls and use a power injector in MRI in a diagnostically helpful way. The simple use of valves inhibits the mixture of contrast media and saline solution. PMID- 9739372 TI - [Magnetization transfer contrast (MTC)--which is the most MTC-sensitive MRI sequence?]. AB - PURPOSE: This study aimed at evaluating MTC-sensitivity of frequently used MR sequences. MATERIAL AND METHODS: 7 sequences (T1-, T2-, PD-weighted SE, four gradient-echo sequences) were combined with a 4 element on-resonance MT-Pulse at 0.5 Tesla (T5, Philips medical systems). The measurements were performed at a knee joint and MT-ratios of cartilage, muscle, fat and a copper sulfate phantom were calculated. RESULTS: Proton density weighted gradient and to a lesser degree spin-echo sequences were superior to T1- and T2-weighted sequences in visualising the MT effects. CONCLUSION: It is important to choose an appropriate MR-sequence for MTC experiments. Proton-density gradient echo sequences are recommended for that purpose. T1- and T2-weighted SE sequences should be avoided. PMID- 9739374 TI - [Radiologic diagnosis of tuberous sclerosis: detection in two generations]. PMID- 9739375 TI - [Two-stage embolization of a planotuberous hemangioma of the neck with thrombogenic microspirals and tissue adhesion]. PMID- 9739376 TI - [Diagnosis and assessment of the course of an intramural hematoma of the thoracic aorta using magnetic resonance tomography]. PMID- 9739377 TI - Conventional X-ray and CT findings in a case of intraluminal choledochocele. PMID- 9739378 TI - [Workshop "Radio-oncology and the Law"]. AB - From 25 to 27 Sept 1997, a workshop was organized at the Essen Medical School (Universitatsklinikum Essen), at which radiooncologists and jurists from universities and courts as well as lawyers contributed their views on mutual problems. The following topics were discussed by papers and in round table meetings: "Requirements on the patient's information", "definition of therapeutic guidelines-limits of clinical research and standard treatments", "treatment documentation", "liability of the physician for treatment faults" and "technical standard and preserve of quality". The consensual guidelines to the topics "patient's information", "therapeutic guidelines" and "liability" are presented here. PMID- 9739379 TI - Hyperfractionated radiotherapy and simultaneous cisplatin for stage-III and -IV carcinomas of the head and neck. Long-term results including functional outcome. AB - PURPOSE: To assess the survival rate, the probability of local control, the patterns of relapse and late sequelae including self-reported quality of life in patients treated with hyperfractionated radiotherapy (RT) and simultaneous CDDP chemotherapy for stage-III to stage-IV carcinomas of the head and neck. METHODS: From 1988 to 1994, 64 patients (median age 55.5 years) with carcinomas of different subsites, excluding the nasopharynx, were treated in a pilot study with 1.2 Gy bid (6 h interval; total dose 74.4 Gy) and simultaneous CDDP (20 mg/m2 daily, 5 days in week 1 and 5) and followed at regular intervals. Overall survival and local control, as well as the rates of late toxicity, were estimated using the actuarial method. Median follow-up was 3.3 years for all and 5.2 years for surviving patients. To assess the quality of life, the EORTC QLQ-C 30 questionnaire and the H&N35 module questionnaire were sent to the patients surviving with no evidence of disease or second primary tumors; they were answered by 15/23 (67%). RESULTS: Overall survival was 37% at 5 years, whereas disease-specific survival was 59%. Twenty-three patients died from uncontrolled head and neck cancer. Second primary tumors were observed in 13 patients, most frequently in the lung. Local control without salvage surgery was 74% at 5 years for all subsites and stages, and loco-regional disease-free survival was 72%. Eleven patients developed distant metastases, which was the only site of failure in 6 cases. Salvage surgery was successful in 2 cases. The actuarial estimates of > or = grade-3 late toxicity was 4% for the mandibular bone and 23% for dysphagia, and 50% of the patients experienced a permanent xerostomy. Self reported global quality of life in surviving patients was good (mean 68 points on a scale 0 to 100); consequences of impaired salivary function had most impact on nutritional and social aspects. CONCLUSIONS: Hyperfractionated RT with concomitant CDDP is well tolerated and highly efficient in controlling moderately advanced to advanced cancers of the head and neck. Second primary tumors are the main cause of death after 3 years and were observed outside of the irradiated area, most frequently in the lung. Even after RT of large volumes to a high dose, salvage surgery can be successfully performed in individual cases. Self-reported quality of life of surviving patients is good, despite xerostomy-associated nutritional difficulties. PMID- 9739380 TI - Conservative treatment of anorectal tumors. AB - PURPOSE: To evaluate the results of interstitial radiotherapy of anorectal tumors. PATIENTS AND METHODS: From 1972 to 1993, one of the authors treated 45 patients by an interstitial implant for anorectal tumors. Of these, 33 patients suffered from primary tumors, 19 from squamous carcinoma, 2 from basaloid carcinoma of the anus and the other 12 from primary adenocarcinoma of the rectum. Of 12 patients treated for local recurrence, 10 had adenocarcinoma and 2 squamous cell carcinoma. Of the 33 patients with primary tumors, 27 received a course of external-beam radiotherapy before the implant. The median follow-up was 35 months. RESULTS: Local response depended on the tumor volume treated. All 21 anal tumors showed complete response, 5 patients developed local recurrence and 4 distant metastases: 3 died from their disease. Of 12 rectal adenocarcinomas, 9 responded completely, 4 patients developed local recurrence and 4 distant metastases; 6 died from active disease. In the last group of 12 patients who were treated for recurrent tumors, 7 responded completely. One patient developed local recurrence and 9 distant metastases, only 4 are alive. CONCLUSIONS: A combination of external-beam and interstitial radiotherapy is a relatively simple, non mutilating, but well-tolerated and very effective method of treatment for early carcinoma of the lower rectum and anus. PMID- 9739381 TI - [Subjective psychological stress and need for psychosocial support in cancer patients during radiotherapy treatment]. AB - PURPOSE: Psychosocial distress and patient attitude towards psychosocial support as well as the correlations with clinical and sociodemographic characteristics should be assessed. METHODS: The stress due to cancer was measured in a consecutive sample of tumor patients at the start of radiotherapy (n = 117) by use of the Hornheide Questionnaire. In addition, the interest of these patients in professional psychosocial support was assessed with the help of the Questionnaire for Psychosocial Support. RESULTS: Patients in the course of radiotherapy and patients with a poor prognosis and advanced disease were more strongly distressed. 32.7% of patients wished professional psychosocial support from the oncologist treating them, 40.6% of the patients wished support from the oncologist and additionally from a psychotherapist or social worker. Interest in professional psychosocial support correlated with the amount of distress, but not with sociodemographic variables. CONCLUSIONS: Results stress the importance of training programs for oncologists in order to improve their ability to detect psychosocial distress in cancer patients and to offer adequate emotional support to them. PMID- 9739382 TI - [Radiation effects in the left kidney after irradiation of the spleen]. AB - BACKGROUND: An important component of treatment of malignant lymphoma is the radiotherapy. If the spleen has to be included in the irradiation field, the left kidney has to be considered as a risk organ. PATIENTS AND METHOD: In 25 patients, splenic pedicle or spleen was included in the irradiation field. These patients were followed up at 6-monthly intervals clinically and by renal scintigram. For 21 out of 25 patients, a volume-dose-histogram of the kidneys was made. RESULTS: A decreased uptake of activity by the left kidney was found in the static renal scintigram of 13 out of the 25 patients and was seen 6 months to 1 year after radiotherapy for the first time at a moderate intensity. The decreased uptake improved in 1 patient, but was progressive in 8 patients until a storage defect or a shrinking of the whole left kidney appeared. The volume-dose-histogram showed that a decreased uptake was seen in the upper half or whole left kidney respectively if at least 40% of the volume or the whole organ was irradiated with at least 20 Gy. 40% of the volume of the left kidney were exposed to at least 20 Gy in only for 3 out of the 12 patients with no decreased uptake. By means of the renal sequence scintigram a reduced function of the left kidney was determined for 11 out of 13 patients. The functional contribution of the left kidney deteriorated to 16 to 37% of the total function of the 2 kidneys. One patient developed a hypertension 1 1/2 years after radiotherapy; all other patients showed no clinical symptoms. Retention of substances in blood was not observed. CONCLUSIONS: The static renal scintigram enables defined radiation-induced lesions of parenchyma of the left kidney to be determined after irradiation of the splenic pedicle or spleen. The changes are predominantly subclinical but possible long-term effects are unknown. In the treatment planning all possibilities should be used to minimize the irradiation volume of the left kidney. Furthermore, all patients should be followed up at regular intervals on a long-term basis. PMID- 9739383 TI - [Radiobiological mechanisms for new strategies in radiochemotherapy]. AB - BACKGROUND: The radiobiological mechanisms which may be exploited in the design of new strategies of radiochemotherapy are discussed. METHODS: The identification of exploitable mechanisms of radiochemotherapy has to be based on the analysis of cellular and supracellular mechanisms of tumor cure, acute normal tissue injury and chronic normal tissue injury, respectively, as well as on experimentally documented mechanisms of cellular and supracellular interactions of chemotherapy and radiotherapy. RESULTS: Only the addition of independent cytotoxic effects of radiotherapy and of chemotherapy are a sound basis for predicting the therapeutic effects on tumor cure probability. In the pathogenesis of normal tissue damage, however, complex interactions of supracellular mechanisms play a decisive role which may increase the rate of chronic side effects even in the absence of cellular interactions. CONCLUSION: None of the presently used schedules of radiochemotherapy meets the scientific requirements outlined in the paper. For squamous cell carcinomas the most promising protocol would combine effective doses of chemotherapy and of radiotherapy without dose reduction and/or treatment prolongation simultaneously but without causing increased normal tissue injury. PMID- 9739384 TI - p53 status in radiation-induced soft-tissue sarcomas. AB - BACKGROUND: Following therapeutic irradiation after a latency period of many years radiation-induced tumors, often sarcomas, can arise. Results of radiation induced DNA damage can be 1. p53 over-expression, inducing growth arrest or apoptosis, and 2. occurrence of mutations, frequently including the p53 gene, as one molecular promotor for carcinogenesis. We were interested whether radiation induced sarcomas are associated with alterations of the p53 status. MATERIAL AND METHODS: Samples from 11 radiation-induced soft-tissue sarcomas (STS) were studied by a non-radioactive PCR-SSCP sequencing analysis and by immunohistochemistry with five antibodies for their p53 status. RESULTS: A tumor of one patient possessed a G->A transition in codon 280 (exon 8). Of 11 tumors, 9 showed nuclear p53 positivity, detected by monoclonal antibody DO-1. Of these 9 patients, 7 died during the observation period, whereas the 2 patients with DO-1 negative tumor samples are still alive. CONCLUSIONS: p53 over-expression and p53 mutation occur in radiation-induced STS. p53 status is expected to have prognostic impact for radiation-induced STS. PMID- 9739385 TI - [Dosimetry of a blood irradiator]. AB - BACKGROUND: Blood and blood products are irradiated to avoid the graft-versus host disease (GVHD) in immunosuppressed patients and to destroy tumor cells during the intra-operative autotransfusion in tumor surgery. For that purpose more and more dedicated gamma irradiators are used. In most cases the equipment is supplied with a dose calibration factor for a totally filled irradiation canister. As users handle different blood product volumes, it is necessary to investigate the influence of the irradiated blood volume on the absolute dose in a reference point and the dose distribution in the irradiation volume. MATERIAL AND METHODS: The dose rate in the center of an empty irradiation canister of an IBL 437C blood irradiator (CIS Diagnostic) was investigated by means of Fricke solution dosimeters from the Physikalisch-Technische Bundesanstalt (PTB). Using thermoluminescence dosimetry (TLD) this value could be transferred to a situation with an empty or completely filled respectively with 2 blood samples (270 ml each) filled canister. Also essential for the irradiation of blood is the knowledge of the dose distribution in the irradiated volume. The distributions in the empty and the realistic filled canister were measured by positioning the TLD on the plexiglas holder in a regular pattern. The case of a completely filled container was investigated by means of the MR Fricke gel dosimetry. All distributions are presented as dose-volume-histograms (DVH). RESULTS: The TLD measurement in the center of the completely filled canister yielded a 4.8% higher dose rate value as compared to the suppliers certificate. From the investigations using the Fricke solution dosimeters in air combined with TLD-measurements values for the complete bandwidth of different container fillings could be derived. So the dose rate in the centre of the canister in the boundary conditions empty and full canister as compared to the values for the realistic filling condition (2 bags) are 117.5% and 94% respectively. Axial dose distributions and DVH have been determined for the 3 filling conditions. CONCLUSIONS: We recommend a dose calibration measurement of a blood irradiator to determine the irradiation times for the chosen filling condition, which is typical for the hospital. The DVH presented in this work can be used to derive a value for the dose variance within the irradiated blood. PMID- 9739386 TI - [3D-CT-implanted interstitial HDR brachytherapy + percutaneous radiotherapy and chemotherapy in inoperable pancreatic carcinoma. On the raticle of L. Pfreundner et al. in Strahlenther Onkol 1998;174:133-41 (Nr. 3)]. PMID- 9739387 TI - [Retardation of progression of bone metastasis in mammary carcinoma patients by intravenous pamidronate treatment: results of a randomized, controlled, multinational study]. PMID- 9739388 TI - [Cervix carcinoma, stage IB and IIA without lymph node involvement: the role of adjuvant radiotherapy after radical hysterectomy]. PMID- 9739390 TI - [Value of radiotherapy of residual tumors after chemotherapy of metastatic seminomas]. PMID- 9739391 TI - [Radical surgery or radical radiotherapy in patients with stage Ib and IIa cervix carcinoma]. PMID- 9739392 TI - [Treatment of cerebral metastasis in patients with testicular germ cell tumors]. PMID- 9739393 TI - Innovation to action: marketing occupational therapy. PMID- 9739394 TI - Grip strengths and required forces in accessing everyday containers in a normal population. AB - OBJECTIVE: A commonly adhered to operating principle in occupational therapy clinics is that a person must exhibit 20 lb of grip strength before his or her hand is considered "functional." This study examined the relationship between hand and finger grip performances with the forces required to open common household containers. METHOD: The grip and pinch strengths of 49 college students were obtained using dynamometry. The forces required to open six common household containers were measured using Force Sensing Resistors attached to each container. RESULTS: Weak correlations were found (r = -.179 to r = .333) between grip and pinch strength performances and the forces used to operate the accessing mechanisms of the containers. Analyses of variances demonstrated significantly greater grip and pinch strength performances in men than in women (ps < .05) but no significant difference between the genders in the forces generated to open the containers (ps > .05). CONCLUSIONS: In a normal population of college students, the premise that greater hand strength affords greater performance in accessing everyday household containers was not supported. Implications suggest that grip and pinch dynamometry are not conclusive evaluative tools for predicting hand function while opening a select group of containers. The relationship between traditional dynamometry and hand performance during a variety of functional tasks needs to be examined in clinical populations as well. PMID- 9739395 TI - Clinical interpretation of "grip strengths and required forces in accessing everyday containers in a normal population". PMID- 9739396 TI - Occupational therapists' expectations in rehabilitation following stroke: sources of satisfaction and dissatisfaction. AB - OBJECTIVE: The purpose of this study was to gain understanding of the satisfactions and dissatisfactions in the work of occupational therapists with clients after stroke. METHOD: Data consisted of narrative descriptions by 32 therapists of especially satisfying and dissatisfying experiences in practice. Phenomenology and grounded theory strategies were used for the study design and data analysis. RESULTS: "Expectation" emerged as the core meaning of occupational therapy in stroke rehabilitation. Strong satisfaction was expressed when therapist-informants believed that they had fulfilled their expectations for clients to achieve the following: (a) maximum neuromuscular and functional recovery in affected upper extremities; (b) independence in daily activities; and (c) return to living in the community. Major sources of informants' dissatisfactions were reaching the "plateau" stage of neurological recovery, disagreement between therapist expectations and client and family member expectations, and working with clients perceived as poorly motivated. CONCLUSION: American ideologies about the value of hard work, independence, and self sufficiency appear to strongly shape therapists' expectations, satisfaction, and dissatisfaction in stroke rehabilitation. For occupational therapists, a tension may exist between the idealism of their therapeutic expectations and the realities of stroke recovery as it is experienced within the context of clients' ongoing lives. PMID- 9739397 TI - The relationship of the Allen Cognitive Level Test to demographics, diagnosis, and disposition among psychiatric inpatients. AB - OBJECTIVE: This study examined the relationship of the Allen Cognitive Level Test (ACL) to demographics, diagnosis, and disposition after hospitalization among persons with psychiatric disorders. METHOD: Data were retrospectively collected from medical records, including initial occupational therapy evaluation notes, on 62 female and 38 male patients consecutively admitted to an urban, acute psychiatric inpatient unit. Collected was information on demographics, diagnosis, mental and physical health history, initial ACL scores, role involvement, and discharge living situation (DLS). RESULTS: Patients with higher initial ACL scores were more likely to be younger, to have lived independently before admission, to have been given nonpsychotic diagnoses, and to have been suicidal before admission. Although DLS was most strongly correlated with living situation before admission, ACL scores showed the second strongest correlation to DLS. Patients with higher ACL scores were significantly more likely to be discharged to independent living than were patients with lower ACL scores. CONCLUSIONS: These results provide evidence that patients' cognitive level, as measured by the ACL, may be a useful predictor of community functioning. However, further research is needed to validate the Cognitive Disabilities Model. PMID- 9739399 TI - The effect of autocorrelation on the results of visually analyzing data from single-subject designs. AB - OBJECTIVE: Single-subject research designs are used to conduct clinical research and outcome evaluation in occupational therapy. Confusion exists regarding the best method to analyze and interpret single-subject data. METHOD: One hundred graphs displaying the results of published single-subject research were examined to determine the influence of autocorrelation on the visual inferences made by the original investigators. The graphs were selected from 20 articles published over 10 years in seven rehabilitation journals. The data were extrapolated and lag 1 autocorrelation coefficients computed for both the baseline and treatment phases. RESULTS: Data analysis focused on two issues: (a) whether a relationship existed between the amount of autocorrelation present in a graph and the conclusion on the basis of visual analysis and (b) whether the amount of autocorrelation varied across different phases of the single-subject graphs. When a significant degree of autocorrelation was present, researchers using visual analysis were more likely to conclude that there was no clinically significant change in performance. Autocorrelation values were significantly higher in the treatment phases of the single-subject designs. CONCLUSION: Additional research is needed to establish a set of decision rules to assist clinicians in using visual analysis to evaluate the results of single-subject research. PMID- 9739398 TI - A task for assessing vertigo elicited by repetitive head movements. AB - OBJECTIVE: We developed a simple, inexpensive test for assessing vertigo in persons with peripheral vestibular disorders. METHOD: The test was administered to 16 asymptomatic adults and 16 patients with chronic vertigo caused by peripheral vestibular disorders. Participants sat in a chair and as rapidly as possible transferred 25 beanbags one at a time from a basket placed on the floor to a basket held .91 m up in the air. The task was timed, and the participants rated the level of vertigo elicited on a 10-point scale. RESULTS: Patients took significantly more time to perform the task and reported significantly greater levels of vertigo than did the asymptomatic adults. Test scores did not differ significantly across test sessions or raters. CONCLUSION: Performance on this task reliably differentiated patients with vestibular disorders from asymptomatic adults. The test is inexpensive, takes less than 1 min to perform, and has minimal technical requirements, making it suitable for a variety of facilities and levels of staff expertise. PMID- 9739400 TI - A comparison of two computer access systems for functional text entry. AB - OBJECTIVE: Functional written communication, an important goal in the rehabilitation of persons with tetraplegia, frequently is met through the use of personal computers and alternative computer access systems. To make informed decisions about alternative access systems, the therapist needs information on the efficacy of the available choices. The purpose of this study was to investigate the effectiveness of two commercially available systems for text entry, the traditional mouthstick and the Prentke Romich HeadMaster. METHOD: Participants were a 25-year-old man and 76-year-old woman who both functioned at a C5 neurological level. Neither participant had previous experience with either system for text entry. A single-subject research design was used whereby Participant 1 experienced six phases of treatment (i.e., CBCBCB, where C = mouthstick and B = HeadMaster), and Participant 2 experienced four phases of treatment (i.e., BCBC). RESULTS: Participant 1 achieved a maximum rate of text entry of 5.85 wpm with both the HeadMaster and the mouthstick, whereas Participant 2 achieved a maximum rate of 7.15 wpm with the mouthstick and 4.85 wpm with the HeadMaster. Results from this study were similar to the results from previous comparison studies of persons with severe disabilities who had no experience with alternative computer access systems. CONCLUSION: Both participants were able to use both systems successfully; however, their respective rates of text entry were too slow to be functional in most employment situations. PMID- 9739401 TI - A review of balance instruments for older adults. AB - OBJECTIVE: The purpose of this article is to review balance instruments developed within the past 10 years that can be used in the clinic or home environment. The use of such instruments may assist in identifying older adults who are at risk for falling, a major problem that can result in impaired function and loss of independence. METHOD: Six instruments were reviewed: the Berg Balance Scale (Berg), the Clinical Test of Sensory Interaction and Balance (CTSIB), the Functional Reach Test, the Tinetti Balance Test of the Performance-Oriented Assessment of Mobility Problems (Tinetti), the Timed "Up and Go" Test (TU>), and the Physical Performance Test (PPT). Considered were what aspects of balance are assessed, time needed to administer the instrument, tools or equipment needed, evidence of reliability and validity, advantages and disadvantages, and the target population. RESULTS: The Berg, Tinetti, and PPT measure a variety of aspects of balance, whereas the Functional Reach, TU>, and CTSIB measure more narrow aspects of balance. All six instruments have been used with older adults and do not require much equipment. The instruments differ in their reliability and validity. CONCLUSION: Familiarity with balance instruments can be helpful in selecting the one most appropriate for clinical setting and clients in order to institute appropriate prevention programs, such as environmental modifications and lifestyle adaptations. PMID- 9739402 TI - Play development in children with HIV infection: a pilot study. PMID- 9739403 TI - Tracing research methodology in occupational therapy. AB - As a profession, occupational therapy has been repeatedly confronted with the challenge to prove the value of occupation as a therapeutic medium. The types of research pursued by occupational therapists have evolved in response to societal trends, external pressures, and the priorities of individual practitioners. Although many therapists have reconciled the pursuit of research with the roots of occupational therapy through an adherence to naturalistic methods, others continue to value experimental research designs. This article explores the rise of qualitative research methods in occupational therapy and addresses the current dilemma between naturalistic and positivistic designs. PMID- 9739404 TI - The domain of function: who's got it? Who's competing for it? PMID- 9739405 TI - Cell recovery and appearance in thin-layer preparations in nongynecologic cytology. AB - OBJECTIVE: To compare the diagnostic efficacy of a thin-layer technique with that of a membrane filter technique using a wide variety of fresh cytologic specimens. STUDY DESIGN: Paired samples from 272 nongynecologic cytology specimens were processed for microscopy using thin-layer and membrane filter preparation techniques. Specimens included 162 body fluids and urines, 32 bronchial aspirates and 78 fine needle aspiration biopsies. The two techniques were compared for diagnostic efficacy, cell density/specimen, cell types recovered, qualitative cytomorphology, ease of interpretation and long-term stability of the final preparations. RESULTS: Diagnoses supported by filter preparations were also supported by their matched thin-layer preparations. The same cell types were recovered by both techniques, and the slide cell density was slightly greater using membrane filters. Qualitative morphology and ease of use were superior with the thin-layer samples. After two years, all thin-layer slides were stable and readable, whereas only 43% of the membrane filter slides were usable. CONCLUSION: Thin-layer techniques are morphologically comparable and, in some ways, superior to membrane filter techniques for processing a wide variety of nongynecologic cytology specimens. PMID- 9739406 TI - Transferrin in normal and neoplastic endocervical tissues: distribution and receptor expression. AB - OBJECTIVE: Our recent studies showed that lactoferrin seems to be down-regulated in endocervical adenocarcinomas. We extended those studies to examine the expression of transferrin (Tf) and its receptor (TfR) in endocervical carcinogenesis and any relationship to the expression of lactoferrin, steroid receptors and the proliferative index. STUDY DESIGN: A retrospective study was performed using sections prepared from paraffin-embedded, formalin-fixed surgical specimens of normal endocervix, endocervical adenocarcinoma and cervical adenocarcinoma in situ. Standard immunohistochemical techniques were used to localize Tf and its receptor in the normal and malignant endocervix. In situ detection of mRNA for Tf and the TfR was also performed. The relative intensity of the immunoreaction was estimated using digital computer image analysis. Statistical significance was tested by Student's t test. RESULTS: No differences in the relative staining intensity for Tf and TfR proteins were noted between normal and neoplastic glands. However, quantitation revealed that a greater number of malignant glands stained positive for TfR than observed in the normal endocervix. Expression of Tf and TfR did not correlate with the expression of steroid receptors and lactoferrin or with the rate of proliferation. CONCLUSION: We have demonstrated the expression of Tf and its receptor by both normal and malignant endocervical glands. The increased number of malignant endocervical glands expressing TfR may indicate a special requirement for Tf and the iron that it carries. Our data provide evidence for the existence of a Tf, TfR autocrine or paracrine circuit involved in the regulation of normal and abnormal endocervical physiology. PMID- 9739407 TI - Objective evaluation of Papanicolaou staining. AB - OBJECTIVE: To evaluate the color of cytoplasm stained by Papanicolaou staining quantitatively. STUDY DESIGN: Cytoplasm photographed under the same conditions was measured for spectral reflection by colorimetry, and the color of the cytoplasm was shown by the L*a*b* color specification. RESULTS: This method enabled us to specify the color of cytoplasm, which was classified in light green, eosin and orangeophilic. CONCLUSION: The variation in the color of cytoplasm by different staining methods was quantified and related to the dyes absorbed by the cytoplasm. PMID- 9739408 TI - Cellular morphometry in nongynecologic thin-layer and filter cytologic specimens. AB - OBJECTIVE: To determine the feasibility and utility of thin-layer cytology preparations for morphometric analysis of nuclear and cytoplasmic area in cells from nongynecologic cytology specimens. STUDY DESIGN: Identical paired samples from nongynecologic specimens (bronchial aspirate, urine, pleural and peritoneal fluid) were used to prepare thin-layer preparations and corresponding traditional membrane filter preparations. The paired preparations were analyzed by static image morphometry measuring eight nuclear and cytoplasmic parameters that allowed calculation of nuclear area, cytoplasmic area and total cell size. Hepatocytes and bronchial columnar, mesothelial, squamous and transitional cells were studied, as was a single case of high grade transitional cell carcinoma. Sufficient numbers of each cell type were measured to allow statistical analysis. RESULTS: Both thin-layer and membrane filter preparation techniques yielded individual cells suitable for morphometric analysis, and there were no consistent morphologic measurement differences between the two methods. The thin-layer preparation had the following significant technical advantages: more numerous easily measured single cells, lack of interfering background and superior specimen stability. Benign oval to round nuclei from a variety of cell types have a mean nuclear area in a narrow range from 29 to 55 microns 2. The mean nuclear area of malignant cells studied was significantly larger (78 microns 2), and there was a significant decrease in absolute cytoplasmic profile area in the malignant population studied. CONCLUSION: Thin-layer cytology preparations have significant advantages for morphometric studies over traditional membrane filter cytologic preparations. The morphometric measurement of nuclear area in benign and malignant cell populations has great potential as a generic screening tool for malignancy in cytologic specimens. Simultaneous measurement of cytoplasmic area adds a powerful dimension. The subsequent calculation of a true nuclear/cytoplasmic ratio may yield a sensitive and specific discriminator for detecting malignant cell populations in human nongynecologic cytology specimens. PMID- 9739409 TI - MIB-1 immunohistometry on Tru-cut biopsies in inflammatory and cirrhotic liver disease. AB - OBJECTIVE: To use immunohistometry to analyze the MIB-1 immunopositivity of normal liver parenchyma (n = 25) and of different inflammatory and degenerative liver diseases--including nonspecific inflammation (n = 12), toxic-nutritive (alcoholic) inflammation (n = 12), ascending cholangitis (n = 11), chronic persisting hepatitis (n = 27), chronic-aggressive hepatitis (n = 28) and liver cirrhosis (n = 23). STUDY DESIGN: Immunohistochemical reactions were performed on 3-micron sections of paraffin-embedded Tru-cut biopsies using an indirect peroxidase method. The rate of immunostained cells was determined using the CM-2 TV image analysis system. Twenty viewing fields (0.97 mm2) were measured with 20:1 objective magnification. An average of 2,600 cells were assessed in each case. RESULTS: The values for normal liver tissue were significantly lower as compared with those of all other groups (< .0001 < P < or = .0008). In nonspecific and toxic inflammation and ascending cholangitis, the values did not differ significantly (P > .05). As compared to chronic-persisting viral hepatitis, however, the MIB-1 immunopositivity of nonspecific (P = .0056) and toxic inflammation (P = .0162) was significantly lower. Apart from ascending cholangitis, chronic-aggressive viral hepatitis revealed significantly increased MIB-1 immunopositivity as compared to all other types of inflammation (P < .0001). The MIB-1 values were significantly higher in liver cirrhosis than in the different inflammatory conditions (< .0001 < P < or = .0105) with the exception of chronic-aggressive viral hepatitis. The highest value for normal liver tissue was 0.4%; it was lower than 96% of the values in the different disease groups. Seventy-two percent of the cases with virally induced hepatitis revealed values > 1.5% as compared to only 14% in the other types of inflammation. In chronic aggressive hepatitis and liver cirrhosis, 71% and 65%, respectively, of the values were > 2%. CONCLUSION: MIB-1 immunohistometry allowed the detection of significantly different proliferative activity among different types of inflammatory and degenerative liver diseases, indicating diagnostic value in histologically inconclusive cases. PMID- 9739410 TI - Quantification of eumelanin and pheomelanin: stereologic image analysis method. AB - OBJECTIVE: To develop an ultrastructural stereologic image analysis method allowing quantification of intracellular melanization. STUDY DESIGN: First, in the field of image analysis, a newly elaborated segmentation method, SEM2, was compared with a previously described method based on gray level histograms. Only SEM2 allows the segmentation of melanin in micrographs of poorly melanized melanocytes. Second, in the field of stereology, estimation of cell volume remains problematic in the case of mixed cell populations. This problem is approached by the comparison of stereologic alternatives and cytochemistry (L-3, 4-dihydroxyphenylalanine reaction) in epidermal melanocytes and melanoma cells from several in vitro experiments. The cytochemical approach was found to be the best choice. RESULTS: Concerning the quantification of eumelanin and pheomelanin, the alkali elution method, permitting the specific dissolution of pheomelanin on ultrathin sections, was validated in normal human follicular melanocytes. CONCLUSION: These results allow us to envisage the stereologic quantification of eumelanin and pheomelanin at the ultrastructural level. At present our method is undergoing evaluation by comparison with the present method of reference based on high-performance liquid chromatography. PMID- 9739411 TI - DNA ploidy and morphometric features of normal, hyperplastic, premalignant and malignant specimens: an evaluation. AB - OBJECTIVE: To evaluate nuclear DNA assessments and morphometric variables on various breast cytologic smears and paraffin-embedded tissue sections of human breast and gastric lesions. STUDY DESIGN: Using the "plug or gore" method of Swift and Rasch for absorption cytophotometry with standard optical conditions on the Opton-MPM 01 cytophotometer as well as a graduated eyepiece micrometer on an Amplival microscope, cytologic smears from 33 women and tissue sections from 36 women with breast lesions and 150 patients with various gastric processes were quantitatively evaluated after measurement of nuclear DNA content amounts and morphometric variables. RESULTS: Normal-fitting distributions were found in the diploid-tetraploid regions of the cell cycle. Single cells possessed nuclear DNA content values in the hypodiploid region. Hyperplastic processes--i.e., fibrocystic disease and chronic atrophic (hypertrophic) gastritis--as well as benign tumors (breast fibroadenomas and gastric adenomas) showed abnormally high DNA content: aneuploidy with values in the pentaploid (5C) to octaploid and hyperoctaploid regions. Carcinomas with different origin had DNA values up to 10 16C. We also noted increasing morphometric data in the pathway from normality to cancer. CONCLUSION: Patients with hyperplastic processes who have abnormally high DNA content assessments with aneuploid cell populations and increased morphometric measures have to be very carefully and precisely evaluated. Moreover, these lesions are potentially malignant, with a high risk of cancer development in the future. PMID- 9739412 TI - Automated feature extraction and identification of colon carcinoma. AB - OBJECTIVE: To assess an automated algorithm, developed for the classification of normal and cancerous colonic mucosa, using geometric analysis of features and texture analysis. STUDY DESIGN: Twenty-one images were analyzed, 10 from normal and 11 from cancerous mucosa. The classification was based on a regularity index dependent on shape, object orientation for establishing parallelism and five texture features derived using the co-occurrence image analysis method. RESULTS: Geometric analysis yielded an overall classification accuracy of 80%. The corresponding sensitivity and specificity were 94% and 64%, respectively. Using texture analysis, the overall classification accuracy was 90%, with a sensitivity and specificity of 82% and 100%, respectively. CONCLUSION: This initial study demonstrated that geometric and texture analysis techniques show promise for automated analysis of colon cancer. PMID- 9739413 TI - Remote quantitation server for quality assurance in DNA ploidy analysis. AB - OBJECTIVE: To remotely test a stepwise framework of quality control measures, developed according to the 1997 European Society of Analytical Cellular Pathology consensus on diagnostic DNA image cytometry, in two series of measurements by means of the quantitation server EUROQUANT. STUDY DESIGN: In each of these series, 104 fine needle aspiration biopsies, imprints from breast cancer specimens and 28 rat liver imprints were measured twice at two different cytometry workstations. Further measurements on special rat liver specimens for evaluation of the stability of the machinery and of the preparation process were done at both workstations. Afterwards the measurement data from both machines were transferred to the quantitation server and analyzed automatically. Beside the classical DNA histograms, a set of further evaluations was performed to detect optical errors as well as inhomogeneities in the measurements. Running values for mean integrated optical density values, mean corrective factors and mean coefficients of variations of corrective factors were computed to control the stability of the entire methodology over time. RESULTS: The study demonstrated the preconditions and outcome of server-based quality control in DNA ploidy analysis. The results show that such a remote analysis is feasible and comparable to local DNA ploidy analysis. It is also demonstrated how quality control tests reflect the process performance of ploidy analysis at its different levels. CONCLUSION: Quality control must be an inherent part of the daily routine to allow a reliable diagnostic interpretation and ensure steadily high product quality. PMID- 9739414 TI - Association of prognosis in surgically treated lung cancer patients with cytometric, histometric and ligand histochemical properties: with an emphasis on structural entropy. AB - OBJECTIVE: To explore new tumor features for refined category formation that permits the tailoring of individualized treatment schemes in lung cancer. STUDY DESIGN: Survival data on patients from six independent studies on cases with surgically treated lung cancer, primary lung carcinoids or metastasizing breast carcinoma (including data on primary breast carcinoma) were analyzed by nonhierarchic multivariant discriminant analysis with respect to a set of cytometric/histometric and immunohistochemical/ligand histochemical parameters. The number of stem lines, S-phase-related tumor cell fraction and the extent of structural entropy and its current were measured. In addition, the expression of binding capacities for histo-blood group trisacharides, galectins, the alpha/beta interferon antagonist sarcolectin, the lymphokine macrophage migration inhibitory factor and a monoclonal antibody to the Le(y) epitope was monitored for insight into aspects of immunologic and biologic behavior. RESULTS: In all studies, a correlation between tumor parameters, according to TNM stage and survival, was seen. In order to refine this category formation, at least certain selected features should provide an even more stringent association than TNM stages. Indeed, statistical correlation of the cytometric and histometric parameters as well as the expression of receptors for the two histo-blood group trisaccharides, ligands for the galectins (CL-16, CL-14) and macrophage migration inhibitory factor was stronger than that of TNM stage. A large amount of the current of structural entropy was especially highly significantly associated with poor survival. This observation could be verified in each of the different studies. CONCLUSION: The obtained data strongly support the notion that thermodynamic evaluation of tumor growth focusing on the "entropy distance" of the tumor from its environment is a promising perspective warranting extended studies. Additionally, glycohistochemical features, including binding capacities for histo blood group trisaccharides, have the potential to aid in establishment of a biologic marker set for tumor staging. PMID- 9739415 TI - Possible involvement of proteases in the regulation of spermatogenesis. AB - Proteolytic enzymes, which are synthesized and secreted by cells of the seminiferous tubule of the testis, have important functions in spermatogenesis. We performed metabolic studies using small peptide hormones as a substrate to investigate the activity of proteases in cultured Sertoli cells of the rat. High performance liquid chromatographic analysis of the cell culture supernatants showed cleavage of met- and leu-enkephalin, substance P, and bradykinin. No peptidolysis was observed for the cyclic peptide oxytocin. The hormone cleavage pattern and the use of specific protease inhibitors in peptide degradation experiments demonstrated activities of several proteases in Sertoli cells. These are mainly metalloproteinases including neutral metalloendopeptidases, angiotensin-converting enzyme and aminopeptidases. In addition, activities of serine and aspartic proteases were detected. Only marginal proteolytic activities were observed in Sertoli cell conditioned supernatants, indicating that the investigated proteases are mainly located on Sertoli cell membranes. The peptide hormones used in this study have been found to play a potential role in the endocrine, paracrine or autocrine regulation of testicular cells. The membrane associated proteases reported here may therefore be involved in the metabolism and inactivation of these peptides. PMID- 9739416 TI - Occurrence and formation of cytoskeletal proteins in mammalian spermatozoa. AB - Mammalian spermatozoa are composed of specialized cytoskeletal elements, which appear to have no structural or protein counterparts in somatic cells. Most evident are the outer dense fibres (ODF) and fibrous sheath (FS) of the sperm tail and the perinuclear theca (PT) of the sperm head. The purpose of this study is to review our results on the occurrence and assembly of proteins making up these three elements during spermatogenesis. Our approach was to raise antibodies against the prominent proteins of these elements and to immunolocalize them on testicular sections prepared for histological and ultrastructural analyses. We found that all of the cytoskeletal proteins considered were expressed exclusively during the haploid phase of development and that the proteins of each element had similar if not identical patterns of expression. The PT proteins were synthesized in the first half of spermiogenesis and were associated with acrosome formation, while the ODF and FS proteins were synthesized in the second half of spermiogenesis. The ODF proteins assembled in a proximal-distal direction along the length of the axoneme, while the FS proteins assembled in the opposite direction; both assemblies eventually meeting and overlapping within the periaxonemal cytoplasmic compartment. During assembly the ODF proteins appeared to be temporarily stored in granulated bodies of the cytoplasmic lobe, while the FS proteins were randomly distributed throughout the cytoplasm. In the case of the PT, there appeared to be an interdependence between PT assembly and acrosome formation. The developmental protein distribution patterns observed for each of the elements suggest unique cellular targeting mechanisms adapted by the spermatid to regulate the assemblies of the respective cytoskeletal proteins. PMID- 9739417 TI - Effect of angiotensin converting enzyme (ACE) and angiotensins on human sperm functions. AB - The presence of components of the renin angiotensin system (RAS) and specific receptors of angiotensin II in the female and male reproductive tract supports the hypothesis that reproductive functions may be controlled by RAS. Therefore, the present study investigated the influence of ACE and angiotensins on sperm functions and the sperm-egg interaction. The experiments did not indicate direct effects of ACE on the capacitation process or acrosome reaction. Release of ACE from human spermatozoa during capacitation was not related to their ability to undergo acrosome reaction after stimulation with ionophore. Therefore, ACE release does not seem to be a useful clinical marker for human sperm capacitation. However, decreased binding of human spermatozoa to the oolemma of zonafree hamster oocytes after inhibition of ACE by captopril indicates that kininase II is involved in sperm-egg interactions. In contrast to other studies, incubation with captopril had no influence on sperm binding to the zona pellucida. Because effects of ACE on sperm-egg interactions but not on capacitation or acrosome reaction were observed, several experiments were performed to study the influence of substrates and products on the acrosome reaction. Angiotensin II induced the acrosome reaction dose-dependently, whereas angiotensin I had no effect on the acrosome reaction. The effect of angiotensin II on acrosome reaction seems to be calcium-dependent and mediated by protein kinases. Since a specific type 2 angiotensin II receptor inhibits the acrosome reaction induced by angiotensin II, this subtype of receptors may be present at the surface of sperm heads. Another clue for the presence of type 2 receptors on human spermatozoa is the finding that pertussis toxin did not inhibit the angiotensin II induced acrosome reaction. In contrast to type 1 angiotensin II receptors, type 2 receptors are known to be G-protein independent. PMID- 9739418 TI - Spermadhesins: a new protein family. Facts, hypotheses and perspectives. AB - Spermadhesins are a novel family of secretory proteins expressed in the male genital tract of pig, horse and bull. They are major products of the seminal plasma and have been found to be peripherally associated to the sperm surface. The structure and function of spermadhesins have been thoroughly investigated in the pig, which exhibits the greatest diversity of members: AWN, AQN-1, AQN-2, PSP I and PSP-II and its glycosylated isoforms. They are multifunctional proteins showing a range of ligand-binding abilities, e.g. carbohydrates, sulfated glycosaminoglycans, phospholipids and protease inhibitors, suggesting that they may be involved in different steps of fertilization. Isolated porcine spermadhesins bind the zona pellucida glycoproteins in a cation-dependent manner with a Kd in a low micromolar range, and AWN, AQN-1 and AQN-3 display similar binding affinity for glycoproteins containing Gal beta(1-3)-GalNAc and Gal beta(1 4)-GlcNAc sequences in O-linked and N-linked oligosaccharides, respectively. During sperm passage through the epididymis AQN-3 and AWN have been shown to bind tightly to the sperm surface by interaction with the phospholipids of the membrane bilayer. At ejaculation the spermadhesins form a protective coat around the sensitive acrosomal region of the sperm head, thus possibly preventing premature acrosome reaction. During in vitro capacitation most of these aggregated sperm adhesins are lost, with the exception of phospholipid-bound spermadhesins. AWN and AQN-3 may now serve as a primary receptor for the oocyte zona pellucida, thus contributing to initial binding and recognition between sperm and egg. The amino acid sequence of spermadhesins does not show any discernible similarity with known carbohydrate recognition domains (CRD). However, they belong to the superfamily of proteins with a CUB domain with a predicted all-beta structure. The crystal structure of the heterodimeric complex of the spermadhesins PSP-I/PSP-II has been solved, showing that the overall structure of both spermadhesins consists of a beta-sandwich with five (parallel and antiparallel) beta-strands. It is the first three-dimensional structure of a zona pellucida-binding protein and reveals the architecture of the CUB domain. The spermadhesins represent a novel class of lectins that may be involved in sequential steps of fertilization, at least in the pig. PMID- 9739420 TI - Contribution of epididymal factors to sperm maturation and storage. AB - In fertile men, the majority of epididymal spermatozoa acquire the potential to fertilize (assessed with sperm function assays) on passage into the corpus and cauda regions of the epididymis. Although secretions of the epididymal epithelium are clearly important for sperm maturation and survival, their role in this process has yet to be fully determined. Alterations in epididymal sperm membranes may result from the incorporation of protein, sugar and lipid determinants. Most probably, factors of epididymal origin act in concert with constitutional changes to spermatozoa, which together permit full sperm function. Epididymal spermatozoa incubated with epididymal epithelial cell cultures can undergo some maturation in vitro, which can lead to the development of sperm fertilizing capacity. Co incubations of human sperm with epididymal epithelial cultures, at 37 degrees C with medium replenished every other day, led to 50% of spermatozoa retaining motility after 8 days. In one case, a few spermatozoa survived for 17 days, the inherent maximal survival time of human spermatozoa in situ. An important aspect of coculture experiments is that close interactions between spermatozoa and epithelial cells can be examined in detail. This coculture technique may yield important information related to epididymal sperm maturation and storage. PMID- 9739419 TI - Function of human epididymal proteins in sperm maturation. AB - Human post-testicular proteins were cloned by subtractive screening of epididymal cDNA libraries, employing testis as the primary negative control. This method identified six human epididymal cDNAs, named HE1-HE6, which are derived from abundant epididymal mRNAs. With the exception of HE5, which turned out to be identical to the lymphocyte surface antigen CD52, they represented completely novel human gene products. To date, there is little information on their function and the mechanism of their deposition on the sperm surface. Unlike the sperm coating antigens, CD52 binds firmly to the sperm membrane via its GPI anchor during epididymal passage. Its synthesis is carefully regulated by the epididymal epithelium. From the results of both in vivo and in vitro studies it was concluded that androgen and temperature are principal factors synergistically modulating epididymal CD52 expression. The human counterparts of two well-known major rodent epididymal proteins, secretory epididymal glutathione peroxidase (sGPX) and acidic epididymal glycoprotein (AEG = Protein DE), were not cloned by the subtractive screening approach, but by RT-PCR amplification. PMID- 9739421 TI - Modulation of mammalian sperm function by fertilization promoting peptide (FPP). AB - Fertilization promoting peptide (FPP; pGlu-Glu-ProNH2) is produced by the prostate gland and secreted into seminal plasma. When added to uncapacitated mouse and human sperm suspensions, it stimulates capacitation as demonstrated by both cytological changes and increased fertilizing ability in vitro. When added to capacitated suspensions, FPP inhibits spontaneous acrosome loss but cells retain high fertility in vitro. Adenosine elicits similar responses to FPP in both uncapacitated and capacitated cells and FPP + adenosine has a greater effect on uncapacitated cells than either used individually. We have proposed that these two molecules modulate the same pathway (adenylate cyclase/cAMP) but act via different receptors. The structure of FPP is crucial for bioactivity: loss of the terminal amide group abolishes activity and substitution of the central glutamic acid can markedly alter activity. Most recently we have found that stimulation of TCP-11, the product of the mouse t-complex gene Tcp-11, elicits responses indistinguishable from those obtained with FPP and we have hypothesized that the protein TCP-11 is the receptor for FPP. The existence of a human homologue for Tcp-11 suggests that the gene product, in conjunction with FPP, could play an important role in human fertility. PMID- 9739422 TI - Evidence for the synthesis and secretion of a CBG-like serpin by human cumulus oophorus and fallopian tubes. AB - The acrosome reaction (AR)-inducing effect of follicular cells, like that of the cumulus oophorus and granulosa cells, has been described previously. In addition to the well known steroid secreting activity of cumulus cells, the results obtained here demonstrate the secretion of a corticosteroid-binding globulin (CBG)-like protein. An AR-inducing effect was shown with the culture medium of human cumulus oophorus. This effect could be eliminated by treating the sample with monoclonal antibodies against CBG. Moreover, Western blotting after SDS-PAGE of the culture medium strongly indicates that human cumulus cells actively express and secrete a CBG-like protein. This might give an indication as to the origin of the acrosome reaction-inducing substance found in follicular fluid. Furthermore, AR-inducing activity and the elimination of this activity by antibodies against CBG was shown for oviductal fluid. With immunohistochemical techniques the CBG-like protein was localized in the epithelial lining of the fallopian tubes, giving possible evidence for the involvement of this molecule in fallopian tube function. PMID- 9739423 TI - News and views of non-genomic progesterone receptors on spermatozoa. PMID- 9739424 TI - New aspects of sperm-zona pellucida binding. AB - Sperm-zona pellucida binding tests have become a widely used diagnostic application for clinicians to obtain guidance in so far as the therapeutic approach of the subfertile couple is concerned. Expanding the oocyte sources is imperative to ensure the consistent use of sperm-zona binding assays. Sources include oocytes derived from post-mortem ovaries, inseminated non-fertilized IVF oocytes and recycled hemizonae. Identification of specific gamete dysfunction is one of the most formidable tasks and fertilization disorders due to defective sperm-zona pellucida interaction are relatively common in the clinical practice, thereby emphasizing the importance of sperm-zona binding tests as diagnostic/predictive tests. Independent publications from Norfolk (USA), Melbourne (Australia), Tygerberg (South Africa) and Israel of highly comparable results confirm that sperm-zona binding tests are good predictors of fertilization. Studies using solubilized human pellucida recently evaluated the influence of solubilized human pellucida on spermatozoa during the capacitational process and subsequent sperm-zona binding. Involvement of G protein and carbohydrate moieties in sperm-zona pellucida binding emphasized the biological and biochemical properties of lectin and have afforded much weight to their employment as membrane probes to evaluate cell surface components. Attention has been focused on the alterations of sperm surface receptors (oligosaccharides) during the differential pathway, epididymal transit and capacitation. PMID- 9739425 TI - Involvement of selectin-like carbohydrate binding specificity in human gamete interaction. AB - The recognition of carbohydrate epitopes by complimentary protein receptors has been shown to be a critical factor in gamete interaction in many different animal species. In this study it was hypothesized that, in the human, gamete binding requires an interaction between selectin ligands on the zona pellucida and putative egg binding proteins on the sperm surface. The hemizona assay (a unique internally controlled bioassay that evaluates tight binding of sperm to the zona) and advanced methods of carbohydrate analysis were used to test this hypothesis. From these tests it was shown that oligosaccharide recognition is also required for initial human gamete binding. This study suggests the existence of distinct egg binding proteins on human sperm that can bind to selectin ligands. Additionally, the results suggest a possible convergence in the types of carbohydrate sequences recognized during initial human gamete binding and immune/inflammatory cell interactions. Glycoconjugates that manifest selectin ligand activity and that express specific carbohydrate epitopes have potent contraceptive and immunosuppressive effects. Such specific oligosaccharide sequences may provide an appropriate recognition signal for embryo development and protection. PMID- 9739426 TI - Zona pellucida as physiological trigger for the induction of acrosome reaction. AB - To evaluate the kinetics of acrosome reaction, sperm samples from four fertile donors were prepared by swim-up and incubated with solutions of human zonae containing 0.1, 0.15, 0.3, 0.5 and 1.0 zonae microliter-1. After 20, 40 and 60 min of incubation at 37 degrees C, aliquots were taken for evaluation of the acrosomal status. The results showed a distinct time- and dose dependence of the acrosome reaction induced by solubilized zona proteins. After 60 min of incubation in 1.0 zonae microliter-1, about 80% of the spermatozoa showed signs of acrosomal loss; about 40% were completely acrosome-reacted. In addition, zona bound sperm showed the same ratios of acrosome-reacted spermatozoa in control experiments. The velocity of acrosome reaction was calculated by means of a double-reciprocal plot being 2.0-2.5% min-1 for completely reacted spermatozoa and those showing signs of acrosome reaction. However, both subgroups differed considerably in their constants of equilibrium (K = 2.0 ZP microliter-1 and K = 0.2 ZP microliter-1, respectively). In nonreacted and partly reacted spermatozoa results might indicate a disturbed course of acrosome reaction or possibly the existence of different subpopulations in respect of sperm competition. PMID- 9739428 TI - Adhesion molecules and matrix proteins on human spermatozoa. AB - Ejaculated spermatozoa and spermatogenic cells express alpha- and beta-chains of beta 1, 3 and 4 integrins as well as their ligands fibronectin and laminin in an extended intra- and interindividual variation and in different patterns of location. The mRNA transcripts of these molecules were detectable by nested polymerase chain reaction in the spermatozoa. The conclusion of a functional competence of these adhesion molecules (AM) was supported by their relation to the results of the zona-free hamster oocyte penetration (HOP) test, the in vitro fertilization of human oocytes and cell attachment assays. AM labelling was influenced by the disintegration of the sperm plasma membrane, especially in seminal plasma, by progesterone, human follicular fluid and microorganisms, but was barely modified by sperm cryopreservation. Despite substantial advances in the knowledge about sperm adhesion molecules, many questions remain to be answered. PMID- 9739427 TI - Immunological identification of zona pellucida 2 (ZP2) protein in human oocytes. AB - The ZP2 protein is a zona pellucida glycoprotein that plays a major role in fertilization. It mediates secondary binding of spermatozoa and is one of the proteins that are involved in zona 'hardening'. ZP2 proteins were identified in various mammalian zonae pellucidae. Their primary structures are highly conserved as revealed by cDNA cloning. Antisera were used against synthetic peptides generated either against a ZP2 amino acid that is homologous in human and mouse ZP2 amino acid sequences (AS ZP2-20) or antibodies against a synthetic human ZP2 peptide (AS ZP2-26). Immunoblots showed that antiserum AS ZP2-20 and AS ZP2-26 strongly recognized human ZP2 protein with an apparent molecular mass of about 72 kDa; both antisera reacted with a minor immunoreactive polypeptide at 96 kDa. In human ovary sections, both antisera revealed immunoreactivity to human zonae pellucidae. Immuno-electron microscopy demonstrated an equal distribution of ZP2 throughout the human zona pellucida. Considerable amounts of immunoreactive material were observed in the ooplasm; some ramification-like extensions of zona pellucida antigen were found close to cells surrounding the oocyte. Our results indicate that antisera against synthetic ZP2 peptides can be used as specific markers for the identification of ZP2 protein in human oocytes. PMID- 9739429 TI - Molecular mechanisms of sperm-egg interactions and egg activation. AB - The binding of acrosome reacted mammalian sperm to the egg plasma membrane initiates a series of signaling events in the egg, termed "egg activation", which lead to the completion of meiosis II and the initiation of a mitotic cell cycle. Many of these signaling events have characteristics of classical signal transduction events in somatic cells. Currently, there are two hypotheses for how sperm-induced egg activation is initiated. In the "receptor" hypothesis, the fertilizing sperm interacts with a specific egg surface receptor, and this interaction leads to signal transduction and effector activation. In the "fusion" hypothesis it is postulated that following fusion of the sperm and egg plasma membranes a soluble sperm-derived factor enters the egg's cytoplasm and activates pathways leading to egg activation. This chapter will provide an overview of the processes of cell-cell interaction and signal transduction leading to mammalian egg activation. It will concentrate on specific molecules proposed to be involved in sperm-egg interaction, signal transduction and effector mechanisms involved in egg activation, and a discussion of sperm-associated factors that have been implicated in egg activation. PMID- 9739430 TI - Therapeutic use of monoclonal antibodies: a new era? PMID- 9739431 TI - Lyon Consensus Conference on high-dose therapy in diffuse large-cell lymphoma. PMID- 9739432 TI - Anthracycline cardiotoxicity, no longer an issue? PMID- 9739433 TI - Current diagnosis and management of medullary thyroid carcinoma. AB - BACKGROUND: Medullary thyroid carcinoma (MTC) originates in the thyroid C cells, accounting for 5% to 10% of all thyroid malignancies. Approximately 75% of cases are sporadic. Significant advances have been made in the molecular biology of MTC, but some aspects of diagnosis and management still remain controversial. DESIGN: We reviewed relevant articles published in major English-language medical journals. We used the MEDLINE database, selected bibliographies, and articles available in our personal files. RESULTS: Mutations of the RET proto-oncogene have been identified in the germline DNA of patients with familial MTC syndromes. Genetic testing can identify patients affected by multiple endocrine neoplasia types IIA and IIB and familial MTC, allowing early diagnosis and possible cure. Surgical treatment is total thyroidectomy. Plasma calcitonin measurements are excellent markers for postoperative follow-up. Adjunctive therapy includes radiotherapy and chemotherapy. The overall prognosis is worse than papillary thyroid carcinoma. CONCLUSIONS: Recent advances in genetic testing allow early diagnosis and treatment of familial MTC syndromes. Despite some advances in treatment, optimal management remains controversial. PMID- 9739434 TI - Systematic reviews of published evidence: miracles or minefields? PMID- 9739435 TI - The use of cardiac biopsy to demonstrate reduced cardiotoxicity in AIDS Kaposi's sarcoma patients treated with pegylated liposomal doxorubicin. AB - BACKGROUND: Pegylated liposomal doxorubicin (PL-DOX) has been shown in preclinical models to induce less cardiotoxicity than non-liposomal doxorubicin. Endomyocardial biopsy is a highly sensitive and specific method for detecting anthracycline-induced cardiac damage. PATIENTS AND METHODS: Myocardial tissue from ten KS patients who had received cumulative PL-DOX (20 mg/m2/biweekly) of 440-840 mg/m2 was evaluated for evidence of anthracycline-induced cardiac damage. Controls were assembled from patients who had received cumulative doxorubicin doses of 174-671 mg/m2 in two earlier cardiac biopsy protocols. Two control groups were selected on the basis of both cumulative (+/- 10 mg/m2) and peak doxorubicin dose (60 or 20 mg/m2, control group 1), or peak dose alone (20 mg/m2, control group 2). RESULTS: PL-DOX patients had significantly lower biopsy scores compared with those of doxorubicin controls despite higher cumulative doses of anthracycline. The median biopsy scores for the PL-DOX and doxorubicin groups, respectively, were 0.3 vs. 3.0 (P = 0.002, Cochran-Mantel-Haenszel row mean difference test) for group 1 and 1.25 for group 2 (P < 0.001, Wilcoxon rank-sum test). CONCLUSIONS: Less severe cardiac changes were seen in patients given PL DOX relative to historical control patients given comparable cumulative doses of doxorubicin. PMID- 9739436 TI - Epidemiology of the non-Hodgkin's lymphomas: distributions of the major subtypes differ by geographic locations. Non-Hodgkin's Lymphoma Classification Project. AB - BACKGROUND: There has been no previous systematic study of the distribution of the major subtypes of non-Hodgkin's lymphoma (NHL) across geographic regions, although there have been isolated reports of such differences. DESIGN: As part of a clinical evaluation of the International Lymphoma Study Group (ILSG) classification of NHL, we classified 1378 NHLs from eight different geographic sites (Omaha, NE, USA; Vancouver, BC, Canada; Capetown, South Africa; London, England; Wurzburg/Gottingen, Germany; Lyon, France; Locarno/Bellinzona, Switzerland; and Hong Kong) using the ILSG classification. RESULTS: Substantial differences were found in the distribution of the major subtypes of NHL across geographic regions (P < 0.0001). A greater percentage of follicular lymphoma was seen in North America, London and Capetown (31% versus 14% at other sites). Peripheral T-cell lymphoma was more common in London, Capetown and Hong Kong (9%) than elsewhere (3%). In Locarno/Bellinzona, higher percentages of mediastinal large B-cell lymphoma (9% versus 2% elsewhere) and mantle cell lymphoma (14% versus 6% elsewhere) were seen. Angiocentric nasal T-/NK-cell lymphoma was only seen in Hong Kong (8%) and Lyon (2%). CONCLUSIONS: Our study provides evidence that the distribution of NHL subtypes differs by geographic region. These findings suggest that geographical differences in etiologic or host factors may be responsible for the observed differences in the distribution of cases across NHL subtypes. PMID- 9739437 TI - Reduced dose of subcutaneous cladribine induces identical response rates but decreased toxicity in pretreated chronic lymphocytic leukaemia. Swiss Group for Clinical Cancer Research (SAKK). AB - PURPOSE: To study the efficacy and the safety of cladribine (2 chlorodeoxyadenosine, 2-CDA) administered as 24-hour infusions or as subcutaneous bolus injections at two different doses to patients with relapsing or refractory chronic lymphocytic leukaemia (CLL). PATIENTS AND METHODS: In this non randomised 2-cohort study, 20 patients with pretreated CLL received cladribine at a dose of 0.7 mg/kg/cycle as continuous i.v. infusions over seven days (group 1) and 35 patients were treated at a reduced dose of 0.5 mg/kg/cycle given as s.c. bolus injections for five days (group 2). After two cycles of four week duration, response was assessed. In the case of progressive disease, therapy was discontinued, otherwise a maximum of four additional cycles were administered until best response. RESULTS: A total of 130 cycles were administered (group 1: 41, group 2: 89). Patient characteristics in both groups were comparable. The median dose intensities were 0.172 mg/kg per week and 0.123 mg/kg per week for groups 1 and 2, respectively (P < or = 0.0001). The overall response rate for all 55 patients was 38% (95% confidence interval (95% CI): 25%-52%), with 5% CR and 33% PR. Response was similar in both patient groups (35% in group 1, 40% in group 2). No association between cladribine dose intensity and response rate was found, and there was no difference between patients relapsing after or refractory to previous therapies (11 of 24 vs. 10 of 31). Median remission duration was six months in both groups. Toxicity, in particular infections (all WHO grades, 34% in group 1 versus 7% in group 2) and myelosuppression (grade 1-4 neutropenia, 72% versus 41% of cladribine cycles) were statistically significantly more frequent in group 1. CONCLUSION: Cladribine is active in heavily pretreated patients with chronic lymphocytic leukaemias. Dose reduction by 29% led to similar response and remission duration, but to a significant decrease of myelotoxicity and risk of infection. Cladribine administered as s.c. bolus injections at 0.5 mg/kg per cycle is safe and this dose level should not be exceeded in this patient population. PMID- 9739438 TI - Randomized trial comparing monthly low-dose leucovorin and fluorouracil bolus with weekly high-dose 48-hour continuous-infusion fluorouracil for advanced colorectal cancer: a Spanish Cooperative Group for Gastrointestinal Tumor Therapy (TTD) study. AB - PURPOSE: The objective of this multicenter study was to compare the efficacy and toxicity profiles of a combination of 5-fluorouracil (5-FU) given by bolus injection together with intravenous leucovorin (LV) versus high-dose 5-FU in continuous infusion (CI) in the treatment of advanced colorectal cancer. PATIENTS AND METHODS: A total of 306 patients were randomized to receive either 5-FU 425 mg/m2 given by bolus injection on days 1-5 plus intravenous (i.v.) LV 20 mg/m2 every four to five weeks or 5-FU 3.5 g/m2/week in a 48-hour CI. Therapy was continued until disease progression. Second-line chemotherapy was allowed in both arms. RESULTS: The response rates in 306 patients with measurable lesions were 19.2% (modulated arm) and 30.3% (CI arm, P < 0.05). The median progression-free survival times were 23.5 weeks (modulated arm) and 25 weeks (CI arm, P = NS). Median survival times were 42.5 weeks (modulated arm) and 48 weeks (CI arm, P = NS). There were no significant differences in grade 3-4 toxicity profiles but if we consider all grades we observed more mucositis in the modulated arm and more hand-foot syndrome in the CI arm. CONCLUSIONS: In terms of response rate, the continuous infusion regimen was more effective than the modulated regimen. There was no significant difference in survival and time to progression, and none in grade 3-4 toxicity. PMID- 9739439 TI - Phase I trial of paclitaxel and gemcitabine administered every two weeks in patients with refractory solid tumors. AB - PURPOSE: Paclitaxel and gemcitabine possess broad spectra of clinical activity, distinct mechanisms of cytotoxicity, and are differentially affected by mutations in cell-cycle regulatory proteins, such as bcl-2. This phase I trial was designed to identify the maximum tolerated dose (MTD) and dose limiting toxicities (DLT) of paclitaxel and gemcitabine when both drugs were given together on a once-every two-week schedule in patients with solid tumors. PATIENTS AND METHODS: A total of 37 patients were treated at nine different dose levels ranging from paclitaxel 75 175 mg/m2 administered over three hours followed by gemcitabinc 1500-3500 mg/m2 administered over 30-60 minutes. Both drugs were administered on day 1 of a 14 day cycle. Dose escalation was performed in a stepwise manner in which the dose of one drug was escalated while the dose of the other drug was kept constant. RESULTS: Dose limiting toxicity (DLT) was observed at dose level 9: paclitaxel 175 mg/m2 and gemcitabine 3500 mg/m2 in the form of grade 4 neutropenia lasting for > or = 5 days (one patient) and grade 3 elevation of alanine aminotransferase (AST/SGPT) (one patient). An analysis of delivered dose intensity (DI) over the first three cycles revealed that higher dosages of both drugs were delivered at dose level 7, paclitaxel 150 mg/m2 and gemcitabine 3000 mg/m2 dose level, than at the MTD, dose level 8, paclitaxel 150 mg/m2 and gemcitabine 3500 mg/m2. Partial responses were confirmed in two patients with transitional cell carcinoma (one of the bladder, one of the renal pelvis) and in one patient with adenocarcinoma of unknown primary. CONCLUSIONS: Paclitaxel and gemcitabine is a promising drug combination that can be administered safely and repetitively on an every-other week schedule. Using this drug administration schedule, the recommended phase II dose is paclitaxel 150 mg/m2 and gemcitabine 3000 mg/m2. PMID- 9739440 TI - Pitfalls in grading severity of chemotherapy-induced peripheral neuropathy. AB - BACKGROUND: Reliable reporting of chemotherapy-induced neurotoxicity is important. The objectives of the current study were to evaluate the differences in the peripheral neurotoxicity sections of several widely used chemotherapy related toxicity grading systems, and the differences in the way in which observers interpret these scales. PATIENTS AND METHODS: Two neurologists independently rated the severity of chemotherapy-induced peripheral neuropathy, according to WHO, ECOG, Ajani, and NCIC-CTC criteria in 37 patients. RESULTS: The highest percentage grade 1, grade 2 and grade 3 peripheral neurotoxicity was noted when employing the WHO, Ajani and NCIC-CTC scales, respectively. Percentage inter-observer agreement across all grades of severity ranged from 45.9 (NCIC CTC) to 83.8 (WHO). The degree of agreement varied from 'poor to fair' to 'substantial'. Percentage inter-observer agreement for the dichotomy grade < or = 2 and grade 3 ranged from 81.1 (NCIC-CTC) to 94.6 (Ajani and WHO), however, exact agreement on grade 3 peripheral neurotoxicity ranged from 0 (Ajani and WHO) to 42% (NCIC-CTC). Percentage interscale agreement for the dichotomy grade < or = 2 and grade 3 varied from 67.6 (WHO and NCIC-CTC) to 100 (WHO and ECOG). Interobserver disagreement of severity grading was partly due to different interpretation of scale parameters. CONCLUSIONS: Our results suggest that caution should be used in interpreting results across studies using different scales for neurotoxicity grading in chemotherapy-related peripheral neuropathy. When (multicentre) trials are to be undertaken with potential neurotoxic or neuroprotective agents, consensus should be sought regarding the toxicity rating scale used, and its interpretation by participating physicians. PMID- 9739441 TI - Quality of life assessment in a cross-cultural context: use of the Rotterdam Symptom Checklist in a multinational randomised trial comparing CMF and Zoladex (Goserlin) treatment in early breast cancer. AB - AIM: The aim of the study is to investigate quality of life (QoL) in the context of a multinational trial. The questions addressed are: is the Rotterdam Symptom Checklist (RSCL) 1) feasible and 2) reliable in cross cultural research, 3) is earlier validation confirmed in a multinational trial and 4) are there systematic differences in QoL across cultures? PATIENTS AND METHODS: Patients with histologically confirmed stage II, node positive breast cancer, were randomised in a multinational trial (the 'ZEBRA-study') comparing standard chemotherapy (CMF) or temporary ovarian ablation by treatment with a LHRH analogue (Zoladex, Goserlin). Patients originating from 13 countries completed a QoL questionnaire at baseline and three months after the start of treatment. RESULTS: 1) The questionnaire was completed by 689 patients at the first and 544 at the second measurement (response 78% and 68% respectively). The proportion of missing data was < 2.5% for 87.8% and 92.7% of the items at the respective time points. 2) Reliabilities of the physical and psychological distress scale were ranging from 0.68 to 0.90 across cultures. Reliability of the activity scale ranged from 0.42 to 0.89. 3) The structure at baseline was in agreement with the two factor structure proposed earlier. 4) Cross-cultural comparison indicated a systematic difference in QoL across cultures (P = 0.0028-< 0.0001) as well as a difference in change across cultures. CONCLUSIONS: QoL assessment using the RSCL proved feasible in the context of multinational clinical trials. Psychometric qualities were satisfactory. Systematic differences in QoL were found between cultures. This finding implies that in multinational clinical trials, treatment comparisons with respect to QoL should carefully account for a differential impact of cultures on the results. PMID- 9739442 TI - Health-related quality of life and sequelae in patients treated with brachytherapy and external beam irradiation for localized prostate cancer. AB - PURPOSE: To evaluate late physical and psychosocial sequelae in patients treated with an association of external beam irradiation (EBI) and brachytherapy (BT) for localized prostate cancer. PATIENTS AND METHODS: Seventy-one patients free of disease, treated at the Centre Francois Baclesse from 1988 to 1992, were enrolled in a case-control study. Seventy-one healthy controls, matched on age and residence, were selected at random from electoral rolls. Two self-administered questionnaires were mailed in January 1996. The French translation of the Nottingham Health Profile questionnaire and the European Organization for Research and Treatment of Cancer (EORTC) QLQ-C30 core questionnaire were used to evaluate physical, role, emotional, cognitive and social functioning, global health status as well as energy and sleep disturbance. Specific problems related to prostate cancer were explored using the prostate specific module developed by the EORTC Genito-Urinary Tract Cancer Cooperative Group. Concordance between clinical complications reported by patients and those reported by physicians was also analyzed. RESULTS: General health quality of life scale scores did not significantly differ between patients and controls, nor did general symptom scale scores. Furthermore, no more late psychosocial sequelae were reported by patients than by controls. No major digestive complications were observed among patients. However, statistical differences were observed concerning interest in sex (P = 0.016) and sexual activity (P < 0.001), urinary incontinence (P < 0.001) and cystitis (P = 0.01). Late subjective morbidity (dysuria, nocturia, urinary incontinence, pelvic pain) appraisal differed slightly between patients and physicians who generally underestimate its severity. While nocturia was reported more often by physicians than by patients (P = 0.0016), patients reported urinary incontinence and pelvic pain more often than physicians (P < 0.001 and P < 0.001, respectively). CONCLUSIONS: The study demonstrates that survivors from localized prostate cancer treated with an association of BT and EBI have good global health status. Major problems that persist are sexual disorders, urinary incontinence and cystitis while digestive disorders were rare. This association could be an alternative to standard EBI in patients with localized prostate cancer. Whatever the treatment choice, patients should be involved in the therapeutic decision which should consider not only expected survival rate but also quality of life. PMID- 9739443 TI - Transferability to clinical practice of the results of controlled clinical trials: the case of antiemetic prophylactic treatment for cancer chemotherapy induced nausea and vomiting. Italian Group for Antiemetic Research. AB - BACKGROUND: There is convincing evidence from randomized clinical trials that the use of 5-HT3 antagonists has brought about a substantial improvement in the control of chemotherapy-induced nausea and vomiting. However, no data exist to indicate how this new research evidence can be applied to the individual patient. PATIENTS AND METHODS: We carried out a prospective, observational study on the use and effectiveness of antiemetic drugs in patients undergoing cancer chemotherapy in 33 Italian oncology departments. RESULTS: A total of 1,956 consecutive patients entered the study; 1,238 of them underwent a one-day chemotherapy and 718 a chemotherapy fractionated over several consecutive days. The 5-HT3 antagonists, used either alone or in combination with a corticosteroid, have almost completely supplanted all other types of antiemetic regimens for preventing cancer chemotherapy-induced emesis. In fact, 80% of patients, irrespective of whether their emesis was acute or delayed, or of the emetogenic potential of the cancer chemotherapy they received, have been treated with these compounds. However, the practice of participating oncologists with respect to prescriptions has been far from consistent with the evidence provided by randomized controlled trials. Both overtreatment and undertreatment have occurred in many patients, creating unjustified costs and placing the patients at greater risk for emesis. However, when antiemetics are properly used their effectiveness is similar to that seen in randomized controlled trials. CONCLUSIONS: Powerful barriers exist between the evidence provided by sound research and clinical practice, and this issue hampers progress toward the optimal use of antiemetic drugs. PMID- 9739444 TI - Totally implantable central venous access ports for long-term chemotherapy. A prospective study analyzing complications and costs of 333 devices with a minimum follow-up of 180 days. AB - BACKGROUND: A few data are available from analyses of the complications and costs of central venous access ports for chemotherapy. This prospective study deals with the complications and global costs of central venous ports connected to a Groshong catheter for deliverance of long-term chemotherapy. PATIENTS AND METHODS: Patients with a variety of solid neoplastic diseases requiring chemotherapy who were undergoing placement of implantable ports over a 30-month period (1 October 1994 to 31 March 1997) have been prospectively studied. Follow up continued until the device was removed or the study was closed (30 September 1997); patients with uneventful implant experience and subsequent follow-ups of less than 180 days were not considered for this study. A single port, constructed of titanium and silicone rubber (Dome Port, Bard Inc., Salt Lake City, USA), was used, connected to an 8 F silastic Groshong catheter tubing (Bard Inc., Salt Lake City, USA). Two-hundred ninety-six devices were placed in the operating room under fluoroscopic control even in the patients treated and monitored in a day hospital setting: 37 of them were in an angiographic suite. A central venous access form was filled in by the operator after the procedure and all ports were followed prospectively for device-related and overall complications. The average purchase cost of the device was obtained from the hospital charges, based on the costs applied during the 30-month period of the study. Insertion and maintenance costs were estimated by obtaining the charges for an average TIAP implant and its subsequent use; the costs of complication management were assessed analytically. The total cost of each device was defined as the purchase cost plus the insertion cost plus the maintenance cost plus the cost of treatment of the complications, if any. The cost of removing the TIAP was also included in the economic analysis when required by the treatment of the complication. RESULTS: Three hundred thirty three devices, for a total of 79,178 days in situ, were placed in 328 patients. Five patients received second devices after removal of the first. In all cases the follow-up was appropriate (median 237 days, range 180-732). Early complications included 10 pneumothoraxes (3.4%; six tube-thoracostomies were applied, 1.8%) and six revisions for port and/or catheter malfunction (overall early complications = 16, 4.48%). Late complications comprised five instances of catheter rupture and embolization (1.5%, 0.063 episodes/1000 days of use), five of venous thrombosis (1.5%, 0.063 episodes/1000 days of use), one of pocket infection (0.3%, 0.012 episodes/1000 days of use), and eight of port-related bacteremia (2.4%, 0.101 episodes/1000 days of use). The infections were caused by coagulase-negative Staphylococcus aureus (five cases), Bacillus subtilis (one case), Streptococcus lactaceae (one case) and an unknown agent (one case); port removal was necessary in six of eight cases. The total cost per patient treated for a six-month period, consisting of the costs of purchase and implantation, treatment of early and late complications, and of maintenance of the device, is US$1,970. CONCLUSIONS: This study represents the largest published series of patients with totally implantable access ports connected to a Groshong catheter. We have shown that US$2,000 are sufficient to cover six months of chemotherapy in one patient using the most expensive commercially available implantable port. According to the present study, totally implantable access ports connected to a Groshong catheter are associated with high purchase and insertion costs, a low complication rate and low maintenance costs. These data support their increasing use in current oncologic medical practice. PMID- 9739445 TI - A primary osteosarcoma of the heart as a cause of recurrent peripheral arterial emboli. AB - The case of a 66-year-old woman with a primary cardiac osteosarcoma is described. These distinctly rare malignant tumors arise preferentially in the left atrium. Clinically, they often present symptoms of both, intramural and intracavitary neoplasm in addition to general weakness, recurrent breast pain, and dyspnea. As shown in the present case, with growing intracavitary tumor masses the risk for peripheral arterial including cerebral embolism increases. Consequently, in most patients with symptoms of systemic arterial embolism of unknown origin performance of transesophageal echocardiography seems advisible, which is presently the most convenient noninvasive imaging method to exclude or to identify intracardiac sources of emboli, irrespective of their type. PMID- 9739446 TI - Treatment and prognosis in a series of primary extranodal lymphomas of the ocular adnexa. AB - BACKGROUND: The aim of this study was to assess clinicopathological characteristics and outcome in a series of primary ocular adnexal lymphomas (POALs). PATIENTS AND METHODS: Nineteen patients with localised (stage IE) POAL were followed for a median of 96 months (24-156). The diagnosis was based on surgical biopsies followed by immunohistochemistry in 16 cases or fine-needle aspiration followed by immunocytophenotypic analysis in three cases. Twelve patients were treated with local radiotherapy (RT), five with chemotherapy (CT), and two refused further therapy after apparently radical tumour removal achieved by the diagnostic excisional biopsy. RESULTS: The histological and immunological pattern was consistent with a diagnosis of MALT-type lymphoma (11 cases), follicular center non-Hodgkin's lymphoma (three cases). a large-cell variant of Burkitt's lymphoma (one case), and large-cell transformed MALT lymphoma (one case). Low-grade lymphoma was diagnosed in the three cases which underwent fine needle aspiration biopsy. All of the patients achieved and maintained complete remission except for those treated with surgical excision alone (two MALT conjunctival lymphoma cases): one of these relapsed locally, the other experienced the systemic spread of a transformed diffuse large-cell lymphoma and died 72 months after diagnosis. The side effects consisted of two cases of RT related cataract after 52 and 72 months. CONCLUSIONS: Regardless of histology, prognosis was excellent when surgery plus RT was adopted, and CT seems to be a valid alternative to RT. Surgery alone may be sub-optimal. PMID- 9739447 TI - Activity of doxorubicin after high-dose ifosfamide in adult patients with advanced soft tissue sarcoma: a study of the Spanish Group for Research on Sarcomas (GEIS). PMID- 9739448 TI - Salvage treatment for germ cell cancer after failed high-dose therapy. PMID- 9739449 TI - HIV-1 reverse transcriptase is capable of elongating derivatives of sequence specific noncomplementary oligodeoxynucleotides. AB - We have carried out a comparison of KM and Vmax values for various primers in the polymerization reaction catalyzed by the HIV-1 RT. The affinity of RT for complementary d(pT)6 containing two different 5'-end pyranone derivatives was 2-3 orders of magnitude higher (KM = 3-15 nM) than that of d(pT)6 (KM = 12.6 mM). Oligodeoxynucleotides (ODNs) noncomplementary to poly(A) template were not elongated by RT. However, derivatives of d(CAGGTG) containing the 5'-terminal chromone and coumarin related groups were efficient primers showing KM (30-300 nM) and Vmax (75-93%) values comparable with that for d(pT)10 (800 nM; 100%). The [d(CAGGTG)]ddT ODN derivatives were effective inhibitors of RT. The primer function of derivatives of noncomplementary ODNs appears to be due to the additional interactions of their 5'-terminal groups with the enzyme tRNA-binding site. PMID- 9739450 TI - Phenotypic differences between diabetes-prone BB rat sublines cosegregate with loci on chromosomes X and 10. AB - BB rat sublines, all developing an insulin-dependent diabetes, differ in several phenotypic traits and in the genotype. To study the genetic basis of phenotypic differences diabetic BB/OK and BB/Mol rats characterized by significantly different frequency and age at onset of diabetes were reciprocally crossed. F1 females of both crosses were backcrossed onto diabetic BB/Mol rats resulting 94 BC1 hybrids which were analyzed for 30 polymorphic microsatellite markers on 14 chromosomes. For the first time it is shown that a diabetes protective locus on chromosome X and a gene around the D10Mit9 locus on chromosome 10 can explain the low frequency (ca. 50%) and the late age at onset of diabetes (ca. 130 days) in the BB/OK rat subline, respectively. PMID- 9739451 TI - Enzymatic methylation of recombinant TIS21 protein-arginine residues. AB - Recombinant TIS21 protein was overexpressed in Escherichia coli harboring the expression vector plasmid pQE-30 carrying the TIS21 cDNA coding sequence containing an extra 120 nucleotides upstream. Employing this protein consisting of 158 amino acid residues of the main chain plus 40 residues of the fusion peptide. It was found that one of the protein methylase I group [S adenosylmethionine:nuclear protein/histone-arginine N-methyltransferase; BC 2.1.1.23; J. Biol. Chem., 269, 1075 (1994)] methylated this protein. The methylation products were identified as guanidino-N-methylated arginines. Some of the kinetics of the reaction are described. PMID- 9739452 TI - Aldosterone stimulates Na,K-ATPase activity in basolateral membrane of rat kidney. AB - The changes of Na,K-ATPase activity and its regulation have been investigated in the renal cortex and its basolateral membrane of aldosterone-induced hypertensive rat. Ouabain-sensitive Na,K-ATPase activity and [3H]ouabain-binding site (Bmax) in the hypertensive rat were significantly increased than those in the control. The levels of Na,K-ATPase alpha 1- and beta 1-subunit mRNA of the renal cortex in hypertensive rat were more increased than those in the control, and their increases were repressed by actinomycin-D. These results suggest that the increase of Na,K-ATPase activities and ouabain binding sites in aldosterone induced hypertensive rat may be correlated with transcriptional regulation of Na,K-ATPase gene expression. PMID- 9739453 TI - Reversible regulation of SHP-1 tyrosine phosphatase activity by oxidation. AB - Increasing evidence indicates that redox regulation is an important signaling mechanism. Protein tyrosine phosphatases (PTPases) are sensitive to oxidative inactivation and are potential targets of redox regulation. In this study, we analyzed the reversibility of oxidative inactivation of the PTPase SHP-1, which negatively regulates protein tyrosine kinase signaling. H2O2 inactivated SHP-1 in vitro. Incubation of the H2O2-inactivated SHP-1 with dithiothreitol recovered 44 99% of the PTPase activity, depending on the H2O2 concentrations used to inactivate SHP-1. Glutathione and N-acetylcysteine also reactivated H2O2-treated SHP-1. Stimulation of SHP-1-transfected HeLa cells with H2O2 rapidly decreased SHP-1 activity, which was completely reversed within 15 min. Thus, oxidative inactivation of SHP-1 is a reversible process. PMID- 9739454 TI - Differential translation of the alpha-1 isoforms of L-type calcium channel in rat brain and other tissues. AB - L-Type voltage operated calcium channel plays an important role in the contraction-relaxation of muscle cells. The alpha-1 subunit of this pentameric protein performs catalytic functions. Multiple mRNA isoforms of this subunit are generated by alternative splicing. For example, an exon encoding 11 amino acids (aa) between the third and fourth transmembrane domains produces two mRNA isoforms in gastrointestinal (GI) tract. The corresponding exon in brain encodes 15 aa. Whether the alpha-1 mRNA isoforms are translated into the corresponding isozymes remain unknown. To address this issue and to characterize the exon in brain, isozymes specific anti-peptide polyclonal antibodies were raised. Both the antibodies reacted with a protein of Mr 180 kDa in the heart and GI-tract, while no reaction was obtained in the kidney or liver. Brain expressed the isoform containing the same exon encoding 11 amino acids present in GI-tract, but the corresponding isozymes were of Mr 145-150 kDa. These findings suggest a tissue specific translation of the alpha-1 isozymes. PMID- 9739456 TI - Fluorescence quenching and time-resolved fluorescence studies on Momordica charantia (bitter gourd) seed lectin. AB - Chemical modification studies implicated tryptophan (Trp) residues in the sugar binding activity of Momordica charantia lectin (MCL) [Mazumdar, T., Gaur, N. & Surolia, A. (1981) Eur. J. Biochem. 113, 463-470]. In the present study, the accessibility and environment of Trp residues in MCL were investigated by intrinsic fluorescence quenching and time-resolved fluorescence. The emission lamda max of native MCL in the absence as well as in the presence of 0.1 M lactose was around 335 nm, which shifted to 365 nm in the presence of 8 M urea, suggesting that the Trp residues which are predominantly buried in the hydrophobic core of the native lectin get exposed to the aqueous environment upon denaturation. At a quencher concentration of 0.5 M, the extent of quenching observed for the native MCL with acrylamide, I- and Cs+ was 46%, 17% and 12%, respectively. In the presence of 0.1 M lactose this quenching was smaller, suggesting that the sugar ligand provides a partial protection to the Trp residues. In time-resolved fluorescence measurements, the decay curves could be fitted well to a biexponential function with the estimated life times 0.92 ns and 4.64 ns for the native protein and 1.15 ns and 5.1 ns in the presence of 0.1 M lactose. All these results are consistent with the involvement of Trp residues in the sugar-binding activity of MCL. PMID- 9739455 TI - Effect of angiotensin convertase inhibitors on lipid peroxidation and peroxyl radical-trapping capacity in rats with experimental diabetes. AB - Effect of two angiotensin convertase inhibitors, enalapril and captopril, on blood plasma and erythrocyte lipid peroxidation and plasma peroxyl radical trapping capacity was studied in rats with streptozotocin-induced diabetes. A progressive increase in blood erythrocyte malondialdehyde (MDA) level was observed in diabetic rats after 6 and 12 weeks. Blood plasma MDA level increased while plasma peroxyl radical-trapping capacity was decreased after 12 weeks. Captopril (2 mg/kg body weight) augmented the diabetes-induced changes in MDA content after 6 weeks and prevented them after 12 weeks increasing also the peroxyl radical-trapping capacity. Enalapril (1 mg/kg body weight) counteracted the diabetes-induced changes in MDA content after both 6 and 12 weeks but did not affect the plasma peroxyl radical-trapping capacity. These results suggest a possibility of a therapeutic use of angiotensin convertase inhibitors to attenuate the effects of oxidative stress in diabetes. PMID- 9739457 TI - Prooxidant character of flavonoid cytotoxicity: structure-activity relationships. AB - The action of flavonoids on bovine leukemia virus-transformed lamb fibroblasts (line FLK) and HL-60 cells was accompanied by lipid peroxidation, their toxicity was partly prevented by iron chelator desferrioxamine and antioxidant N,N' diphenyl-p-phenylene diamine. This pointed out to the involvement of oxidative stress in flavonoid cytotoxicity. The concentration of compound for 50% survival of FLK cells (cL50) did not show correlation with polarographic oxidation half peak potential (Ep/2) and/or partition coefficient (log P) of flavonoids; however, their toxicity to HL-60 cells was described by equation log cL50 (microM) = 3.0161 + 1.1099 Ep/2 (V) - 0.3369 log P. The toxicity of quercetin was partly prevented by nontoxic concentrations of other flavonoids examined, thus pointing out to potential neutralization of quercetin cytotoxicity by intake of flavonoid mixtures. PMID- 9739458 TI - RAPD profile variation amongst provenances of neem. AB - Neem, described as a tree for solving global problems, is an evergreen, long lived, multipurpose tree of the tropics with a wide distribution range in India. It is believed to be highly cross-pollinated. Inter-provenance variations have been reported in neem in case of morphological and physiological characters. Yet no reports about the genetic determinism for these variations are available to our knowledge. In order to have an idea about the extent and/or nature of genetic (DNA) variation in neem, the powerful RAPD technique has been employed. RAPD profiles of 34 accessions/provenances of neem were generated with 200 decamer random primers, of which the data from the 49 primers, that resulted in reproducible amplification products, were considered for analysis. Based on the presence/absence of bands, a similarity matrix was computed. Dendrogram was constructed by UPGMA method based on the pairwise similarities amongst the RAPD profiles. The similarities in RAPD profiles amongst the different DNAs was more than that expected due to the cross-pollinated nature of the tree and furthermore, these more-than-expected similarities were not due to random chance. These results suggest that neem may have a narrow genetic base. PMID- 9739459 TI - Inactivation of creatine kinase is due to the conformational changes of the active sites during thermal denaturation. AB - The conformational changes of the active site of creatine kinase (ATP: creatine N phosphotransferase EC 2.7.3.2.) during thermal denaturation was followed by changes in fluorescence at the active site of the enzyme labeled by o phthalaldehyde. Conformational changes of the active site occurred at the same time as inactivation of the enzyme. The active site changes occurred before the denaturation of the enzyme molecule as a whole was detected. The above results showed that the thermal inactivation of the creatine kinase was due to the conformational changes of its active sites. PMID- 9739460 TI - Kinetics of inactivation of Penaeus penicillatus acid phosphatase during inhibition by N-bromosuccinimide. AB - In the present investigation, the inactivation by N-bromosuccinimide of acid phosphatase from penaeus penicillatus has been studied using the kinetic method of the substrate reaction during modification of enzyme activity as previously described by Tsou [(1988, Adv. Enzymemol. Related Areas Mol. Biol. 61, 381-436]. The results show that inactivation of the enzyme by N-bromosuccinimide is a slow, reversible reaction. The results also clearly show that the modification of the tryptophan residues of penaeus penicillatus acid phosphatase by high concentrations of N-bromosuccinimide led to the complete inactivation of the enzyme. The microscopic rate constants were determined for the reaction of the inactivator with the free enzyme and with the enzyme-substrate complex. Comparison of the obtained microscopic rate constants indicates that the presence of the substrate offers marked protection of the enzyme against inactivation by N bromosuccinimide. The above results suggest that the tryptophan residue is essential for activity and may be situated at the active site of the enzyme. PMID- 9739461 TI - Modelling cortical cataractogenesis. XXIX. Calpain proteolysis of lens fodrin in cataract. AB - The relation between cataract and calpain proteolysis of lens fodrin was studied in two systems: elevated glucose (55.6 mM, diabetic model), and cytochalasin D (CD, 10(-2) mM, actin depolymerization-induced opacity model). Glucose treatment (48 h) caused a visible opaque layer and enzyme leakage, with a concomitant accumulation of ([Ca2+]i) around the lens equatorial cortex. CD caused both earlier and greater opacity and enzyme leakage than glucose. Lens fodrin digestion occurred in parallel with the timing and extent of calcium elevation. A calpain inhibitor peptide (CIP, 10(-2) mM) reduced the proteolysis of fodrin, opacity, and enzyme leakage in glucose-treated lenses but only partially retarded them in CD-treated lenses. These results suggest a mechanism in which calpain proteolysis of fodrin is a critical event in lens damage during opacification of cortical cataract. PMID- 9739462 TI - Nucleotide sequencing of S-RNA segment and sequence analysis of the nucleocapsid protein gene of the newly isolated Akabane virus PT-17 strain. AB - The nucleotide sequences of the S-RNA of Akabane viruses JaGAr-39, OBE-1, Iriki and the newly isolated PT-17 strains and the Aino virus were determined and compared. The results reveal that the S-RNAs of the four Akabane strains share 96.9% homology in nucleotide sequences. Only one amino acid difference out of the 233 amino acids of the nucleocapsid protein (N) and three amino acid differences in the 91 amino acids of the nonstructural protein (NSs) were found among the Akabane viruses. Amino acid sequences of N and NSs proteins of the Aino virus have approximately 80% identity as compared with the Akabane viruses. The results also demonstrate that the four Akabane viruses and the Aino virus can be clearly differentiated by RFLP (restriction fragments length polymorphism) analysis using RT-PCR generated nucleocapsid protein genes and digested with HaeIII and HindIII. The phylogenetic tree based on the UPGMA (Unweighted Pair Group Method with Arithmetic Mean) analysis of the sequences of nucleocapsid protein genes and the S-DNAs revealed that the newly isolated PT-17 strain is most closely related to Iriki strain, than the JaGAr-39 or OBE-1 strains. PMID- 9739463 TI - Purification and cell attachment activity of a D-galactose-binding lectin from the skin of sea hare, Aplysia kurodai. AB - A D-galactose-binding lectin which does not require Ca2+ or reducing reagents to induce activity was purified from skin of sea hare, Aplysia kurodai, by affinity chromatography. Skin lectin was confirmed to be a disulfide-bonded heteromultimer with molecular mass of 200 kDa, consisting of 28 kDa and 26 kDa subunits by SDS PAGE and two-dimensional SDS-PAGE. Human rhabdomyosarcoma cells attached to and spread on plastic plates coated with lectin. Cell adhesion induced by lectin was completely inhibited by the addition of lactose. PMID- 9739464 TI - Molecular organization of the gene encoding Xenopus laevis transforming growth factor-beta 5. AB - Transforming growth factor-beta s (TGF-beta s) are multifunctional polypeptides, known to influence proliferation and differentiation of many cell types. TGF-beta 5 cDNA was cloned from Xenopus laevis and this isoform is unique to the amphibians. Here, we report the isolation and characterization of the TGF-beta 5 genomic clones to determine the structure of TGF-beta 5 gene. The gene consists of seven exons and all intron-exon boundaries follow the GT-AG consensus. The organization of TGF-beta 5 gene was identical to that of the mammalian TGF-beta isoforms, with the exception of position of the first splice junction. We determined the size of TGF-beta 5 gene to be approximately 20 kb. PMID- 9739465 TI - Modulation of HPV16 E7 translation by tRNAs in eukaryotic cell-free translation systems. AB - Translational regulation of HPV16 E7 mRNA is of particular interest since the viral E7 protein has oncogenic activity. Here we report that additional supplementation of cell-free reticulocyte/wheat-germ translation systems with rat liver tRNA pool favors the expression of the viral oncoprotein E7 which otherwise is scarcely translatable. The translational activation is explained by the positive correlation between the frequency of the glutamic and aspartic codons in E7 mRNA, and the increased concentration of the corresponding tRNAs following tRNA supplementation. The present in vitro data suggest a link between E7 expression and cell conditions conductive to tRNA changes such as development, cell proliferation and aging. PMID- 9739466 TI - Biochemical characterization of the third domain from Bacillus thuringiensis Cry1A toxins. AB - Cry proteins from Bacillus thuringiensis have insecticidal properties. The function of domains I and II has been described but domain III has so far eluded understanding. Domain III from Cry1Ab and Cry1Ac has been cloned, expressed in E. coli and injected to rabbits with the aid of characterizing them immunologically. Interestingly, polyclonal antibodies against Cry1Ab fragment did not recognize either the native Cry1Ab toxin or the Cry1Ac fragment while those against the latter did recognize either the native Cry1Ac toxin or the Cry1Ab protein fragment. A combination of information from sequence comparison and hydrophobicity profile indicates that these protein fragments possibly adopt different spatial dispositions within the respective toxins. PMID- 9739467 TI - Physico-chemical properties of copper-oxidized very low density lipoproteins. AB - The aim of our study was to investigate the effect of Cu2+ catalyzed oxidation on VLDL physico-chemical properties and secondary structure of apo B-100. Incubation of very low density lipoproteins with copper ions resulted in a decrease in tryptophan and lysine residues parallel to lipid peroxidation products, conjugated dienes and TBARS. Fluorescence polarization showed an increase in the molecular order at the lipoprotein surface of VLDL, as demonstrated by the increase in Pf values of DPH. The secondary structure of apo B-100 was investigated by infrared spectroscopy. Increased order and structural changes, as observed after oxidative stress on VLDL, could be of relevance in the abnormal interactions between lipoproteins and cell membranes. PMID- 9739468 TI - O2-binding to fully reduced cytochrome aa3 reexamined using SVD analysis of simulated and real data. AB - Using new time-resolved multichannel experimental and simulated data, analyzed by SVD, it is concluded that the second order binding rate constant for O2-binding to fully reduced mammalian cytochrome aa3 is approximately five times higher than the currently accepted value of approximately 1.5 x 10(8) M-1 s-1. PMID- 9739469 TI - Cytochrome c oxidase from eucaryotes but not from procaryotes is allosterically inhibited by ATP. AB - The activity of reconstituted cytochrome c oxidase from bovine heart but not from Rhodobacter sphaeroides is allosterically inhibited by intraliposomal ATP, which binds to subunit IV. The activity of cytochrome c oxidase of wild-type yeast and of a subunit VIa-deleted yeast mutant, measured with Tween 20-solubilized mitochondria in the presence of an ATP-regenerating system, was also allosterically inhibited by ATP, indicating the general validity of this mechanism of "respiratory control" in eucaryotic cytochrome c oxidases (Arnold and Kadenbach, Eur. J. Biochem. (1997) 249, 350-354). Deletion of subunit VIa changes the biphysic into monophysic kinetics of the yeast enzyme in the presence of ADP. A tenfold higher amount of horse heart cytochrome c, as compared to yeast cytochrome c, was required to relieve the ATP inhibition of the yeast enzyme. PMID- 9739470 TI - Immunological detection of ecto-ATPase in chicken and rat tissues: characterization, distribution, and a cautionary note. AB - We have generated a polyclonal antibody (CKG2) against native chicken gizzard ecto-ATPase for immunolocalization and immunoprecipitation. Active ecto-ATPase is immunoprecipitated from solubilized chicken and rat membranes and shown to be localized to the plasma membrane of the chicken smooth muscle cells. This antibody is specific for the ecto-ATPases, since the more abundant chicken stomach ecto-apyrase is not recognized in immunoprecipitation, western blot or immunolocalization analyses. The CKG2 antibody cross-reacts with mammalian (rat) ecto-ATPase in western blots, with testis being the most abundant source. Interestingly, when the same rat membranes are analyzed by western blot under non reducing conditions, the 66 kDa ecto-ATPase is not recognized, instead a 200 kDa protein is detected, previously postulated to be an oligomer of ecto-ATPase. However, this 200 kDa cross-reacting protein is not related to the ecto-ATPases, but is instead an immunoglobulin binding protein, comprised of 50 kDa subunits. PMID- 9739471 TI - Acute-phase induced phosphorylation of rat liver nucleoprotein p70 modulates its binding affinity for the haptoglobin gene. AB - An increase in the binding affinity of rat liver trans-acting nucleoprotein p70 for the hormone responsive element of the rat haptoglobin gene in acute-phase reactions has implicated a posttranslational modification. This investigation examines the proposed acute-phase related structural alterations of p70 using an in vitro phosphorylation/dephosphorylation assay and selective digestion of p70 with Staphylococcal aureus V8 protease. The results show that p70 requires phosphorylation to express its DNA-binding ability. Selective proteolysis of p70 provided evidence that acute-phase induced phosphorylation of this protein alters its conformation in such a way that its DNA-binding ability is increased. PMID- 9739472 TI - Safety investigation: interaction of infant radiant warmers and bilirubin phototherapy lights in the regulation of temperature of newborn infants. PMID- 9739473 TI - Service program diagnostics and decision support technology for improving health technology service efficiency and productivity. AB - Our in-depth survey of both the state of the art and the results of application, implementation, and rollout of problem resolution diagnostics in control help desks and field service clearly shows the potential of online remote diagnostics technology (Figure 9) to improve service force productivity and efficiency and to make more effective use of service resources (people and parts). Our survey of the current and planned future expenditures for diagnostics used in field service clearly indicates that the overall service diagnostics market is currently sizable, in the range of $2 billion as of 1997, and is projected to grow to approximately $2.6 billion by 2000. This current expenditure and growth will occur primarily in the electronics arena, but there are still substantial development and application investments occurring in both electromechanical and mechanical areas. We believe that the pace of investment will fall off as the service industry shifts from development and experimental research to the application and rollout of standard off-the-shelf technology. Thus, the overall pattern of the diagnostics market indicates a continuing increase in expenditures by the field service industry for electronics-oriented service diagnostics and some increase with respect to use of this technology in electromechanical and mechanical applications. The application and use of diagnostics technology will also be affected by the ability of the field service management community to recognize the need to search for an optimum as opposed to a feasible solution to dispatch, assignment, call screening, call avoidance, and logistics deployment. The further rollout of affordable wireless communications technology such as Ardis and Ram mobile and the new cellular-based CPDP technology, coupled with the increased availability of inexpensive wireless-based laptops, portables, and CD ROM-based problem resolution and diagnostics technology clearly shows the value of this technology in improving service productivity, efficiency, and profitability. As we move from very expensive academically and research and development-oriented diagnostics applications to the purchase and use of off-the shelf software and predeveloped knowledge bases, it is clear that this technology will be broadly applied. PMID- 9739474 TI - A potentially powerful new tool for noninvasive diagnosis of cardiac abnormalities: the CUPID system for analysis of electrocardiograms in the frequency domain. AB - Clinical correlation of frequency-domain characteristics of electrocardiograms with specific disease entities of thousands of patients has produced a simple, noninvasive diagnostic system capable of detecting many cardiac abnormalities with specificity and sensitivity exceeding 90%. This system, a proprietary product called CUPID, is now undergoing clinical trials in Europe, South America, and the United States. The author discusses the physiologic basis of electrocardiography and the development of the CUPID system, and reviews the results of the trials that have been completed. The system appears to hold promise for fast, inexpensive, and accurate screening of patients for cardiac disease without somatic invasion. PMID- 9739475 TI - Electroventilation--a missed opportunity? AB - "Electroventilation" identifies the many techniques used to produce artificial respiration by the application of trains of stimuli to strategically placed body surface electrodes. The author reviews the efficacy and safety of electroventilation and traces its history and present status. The theoretical and practical bases for the method are presented, along with numerous examples of its use in man and animals. The article concludes with the indications and contraindications for electroventilation. PMID- 9739476 TI - Implantable cardiac assist devices. PMID- 9739477 TI - Reengineering clinical engineering. PMID- 9739478 TI - Chronopharmacokinetics. Current status. AB - Absorption, distribution, metabolism and elimination are influenced by many different physiological functions of the body which may vary with time of day. Thus, the pharmacokinetic parameters characterising these different steps, conventionally considered to be constant in time, depend on the moment of drug administration. Time of day has to be regarded as an additional variable influencing the kinetics of a drug. Chronokinetic studies have been reported for many drugs in an attempt to explain chronopharmacodynamic phenomena and demonstrate that the time of administration is a possible factor of variation in the kinetics of a drug. In this paper this is illustrated with the chronopharmacokinetics of cardiovascular and nonsteroidal anti-inflammatory drugs. Time-dependent changes in kinetics may proceed from circadian variations at each step, e.g. absorption, distribution, metabolism and elimination. Thus, circadian variations in gastric acid secretion and pH, motility, gastric emptying time, gastrointestinal blood flow, drug protein binding, liver enzyme activity and/or hepatic blood flow, glomerular filtration, renal blood flow, urinary pH and tubular resorption may play a role in such kinetic variations. New tools, such as new formulation procedures or pumps with constant or programmable delivery rates, now make it possible to deliver a drug at a definite time, or during a definite span of time and at a controlled rate in chronokinetic studies. Microdialysis would be of particular interest in chronopharmacological and chronokinetics studies, but, surprisingly, very few chronobiological studies have been conducted using this technique. With regard to new models and concepts in chronokinetics, pharmacokinetic-pharmacodynamic modelling may be useful, and modelling of the chronopharmacological patterns of drug response and kinetics have been attempted by some authors. Drug chronopharmacokinetic knowledge may be clinically relevant as it may have implications for drug prescription by modulating the distribution of the total daily dose along the 24-hour scale. However, it seems reasonable to consider chronopharmacokinetic studies in specific cases related to patients, illness or the drug itself. When conducting a chronokinetic study it is not only necessary to take into account differences in the time of administration but to also have strict control of all other possible variables which are known to influence pharmacokinetic processes. PMID- 9739481 TI - Methodological issues in pharmacokinetic-pharmacodynamic modelling. AB - This article presents the theoretical and practical aspects involved in the design and analysis of pharmacokinetic-pharmacodynamic modelling studies. The main features of the protocol of pharmacokinetic-pharmacodynamic studies are discussed with special focus on experimental designs in relation to individual and population approaches. Some basic pharmacodynamic models (such as linear, log linear, hyperbolic and sigmoid models) are presented as well as more complex time dependent models (effect compartment and physiological indirect response, tolerance models) which are required when the concentration-effect relationship shows a hysteresis loop. The methods of estimation, with special focus on the individual and populations approaches, are covered, along with the way pharmacodynamic models and methods of estimation can be applied to real data and the information required to criticise the results of modelling. We also present some real problems frequently encountered when performing pharmacokinetic pharmacodynamic modelling and give some potential solutions (problems with hysteresis loops, lack of convergence, problems with residuals). The last section discusses the significance of pharmacodynamic parameters. PMID- 9739483 TI - Leptin, the hypothalamus and the regulation of adiposity. AB - The hypothalamus contains a wealth of peptide and non-peptide neurotransmitters, many of which have been shown experimentally to influence feeding behaviour and energy metabolism. Regulatory activities as complex as these are likely to be controlled by numerous neurotransmitters interacting at a variety of levels. The hierarchy of command of these neuronal circuits is not known, but it is possible that some converge on to a final common pathway, governed by the actions of a single neurotransmitter, through which all other influences ultimately operate. This review will discuss several of the more recently identified neurotransmitters and consider their validity as candidates in the regulation of energy homeostasis. PMID- 9739482 TI - Lipid-lowering drugs: who gets what? PMID- 9739479 TI - Pharmacodynamics and pharmacokinetics of thiopental. AB - Thiopental is an ultra short-acting barbiturate which remains the standard against which other induction agents are judged; it is also indicated for the therapy of brain hypoxic-ischaemia injuries and status epilepticus. Aspects of drug distribution that govern the onset and end of drug effect have been intensively studied to determine which parameters (in patient characteristics, diseases and administration modalities) influence effective dose and concentrations in individual patients. Thiopental has been used as a reference for pharmacokinetic and/or pharmacodynamic models in the study of rapid and short acting effect drugs. In anaesthesiology the pharmacokinetics of thiopental are described as linear; when doses and duration of treatment increase, nonlinear pharmacokinetics occur because of the saturation and/or the induction of the metabolism. PMID- 9739484 TI - The role of dietary fatty acids in lipoprotein oxidation and atherosclerosis. AB - Although it is well established that dietary saturated fatty acid intake is an important risk factor for coronary heart disease, there remains substantial controversy regarding whether these dietary fatty acids should be replaced with either carbohydrates, monounsaturated fatty acids, polyunsaturated fatty acids or a combination of these. This review highlights recent studies evaluating the role of dietary fatty acids in atherosclerosis, with a particular emphasis on their roles in lipoprotein oxidation and other potential proatherogenic processes. PMID- 9739480 TI - Pharmacokinetic drug interactions with anti-ulcer drugs. AB - The safety profile of any pharmacological agent is defined on the basis of its toxicity, tolerability and potential for pharmacokinetic and/or pharmacodynamic interactions with other compounds, which may belong to the same or to a different pharmacological class. Drug-drug interactions are important in clinical practice because short and long term therapeutic regimens frequently require coadministration of different drugs. The pharmacological treatment of gastric and duodenal ulcers (and of related syndromes) includes older and newer compounds, which have different mechanisms of action and exert different therapeutic effects. These compounds are widely prescribed in combination with other drugs being given for the treatment of concomitant diseases. This article reviews pharmacokinetic interactions with anti-ulcer drugs, paying particular attention to those which have clinically relevant adverse effects. Drugs mentioned in the literature as causing any pharmacokinetic interaction with anti-ulcer compounds are considered in this article. PMID- 9739485 TI - Low birth weight and cardiovascular disease: myth or reality? AB - The Barker hypothesis proposes a link between low birth weight and adulthood cardiovascular disease. Recent studies have cast doubt on this hypothesis whilst others have proposed an effect of maternal birth weight that may extend to future generations. Thus, the debate on the effect of birth weight in cardiovascular disease continues. PMID- 9739486 TI - Genotype/phenotype correlations in familial hypercholesterolaemia. AB - It is now possible to identify the specific gene defect in the majority of patients with familial hypercholesterolaemia. A potential benefit of this knowledge, in addition to helping with family screens, is to be able to predict the future clinical course. In order to do this, detailed genotype/phenotype correlation studies are required. PMID- 9739488 TI - Are risk factors for stroke and coronary disease the same? AB - Many risk factors operate in both coronary heart disease and stroke, especially ischaemic stroke--age, sex, social class, blood pressure, pre-existing vascular disease (angina, myocardial infarction, cardiac failure, diabetes and peripheral vascular disease, transient ischaemic attack and stroke), atrial fibrillation and fibrinogen, smoking, alcohol and height. Total cholesterol has also recently been recruited to this list. The various mechanisms involved in stroke and its subtypes and the epidemiological problems in evaluating aetiological factors in stroke make the comparison with coronary heart disease more difficult. The recent discrepancy between much of the epidemiology and the clinical trials evaluating the role of lipids in stroke has spurred the systematic review (meta-analysis) of major prospective observational studies. These will provide a clearer assessment about the quantitative comparison of some of the more important risk factors for stroke and coronary heart disease in the near future. PMID- 9739487 TI - Paraoxonase and coronary heart disease. AB - Paraoxonase (PON1) hydrolyses organophosphate insecticides and nerve gases and is responsible for determining the selective toxicity of these compounds in mammals. Human PON1 has two genetic polymorphisms giving rise to amino-acid substitutions at positions 55 and 192. The 192 polymorphism is the major determinant of the PON1 activity polymorphism towards organophosphates. However, the 55 polymorphism also modulates activity. Ex vivo, the PON1 polymorphisms are important in determining the capacity of HDL to protect LDL against oxidative modification in vitro and this may explain the relationship between the PON1 alleles and coronary heart disease in case-control studies. In recent case-control studies serum PON1 concentration and activity were also found to be decreased in coronary heart disease (CHD) independent of the PON1 polymorphism, and in diabetes serum PON1 specific activity decrease is also independent of the PON1 genetic polymorphism. HDL from transgenic mice lacking PON1 fails to protect LDL against oxidative modification. Thus PON1 may be a determinant of resistance to the development of atherosclerosis by protecting lipoproteins against oxidative modification, perhaps by hydrolysing phospholipid and cholesteryl-ester hydroperoxides. PMID- 9739489 TI - Lipoprotein classes and coronary disease regression. AB - Lowering LDL cholesterol (LDL-C) levels to reduce or prevent coronary artery disease (CAD) progression and cardiac events in hypercholesterolemic subjects is now widely accepted. The clinical benefit of lowering LDL-C has recently been extended to individuals with normal or mildly elevated LDL-C. Recent analyses of large primary and secondary CAD prevention trials, however, clearly demonstrated that reducing LDL-C levels does not entirely account for the coronary event reduction associated with lipid-lowering therapy. Growing and compelling evidence is emerging on the role of triglyceride-rich lipoproteins (VLDL and IDL), high density lipoproteins (HDL), and small, dense LDL, as well as non lipid risk factors, in the regression or stabilization of atherosclerotic plaques of mild/moderate severity, which are associated with clinical cardiac events. Enzymes involved in the tight metabolic interrelationship between triglyceride rich lipoproteins, small, dense LDL and HDL levels may represent potential therapeutic targets for CAD prevention by favourably altering lipoprotein composition and physical properties in addition to the current therapeutic focus on lipoprotein levels. PMID- 9739490 TI - Is oxidized low-density lipoprotein present in vivo? AB - Basic research has provided strong evidence that oxidation of LDL plays an important role in the pathogenesis of atherosclerosis. Several mechanisms have been identified which can lead to LDL oxidation in vivo. Clinical and epidemiological studies have provided circumstantial evidence that oxidized LDL, as measured by serum autoantibody levels, may be associated with the progression of atherosclerotic vascular disease. This review discusses recent findings regarding the presence of oxidized LDL (ox-LDL) in vivo and the significance of ox-LDL autoantibody measurements as a tool to predict cardiovascular diseases in various patient populations. PMID- 9739491 TI - Hyperlipidaemia and cardiovascular disease. PMID- 9739492 TI - Nutrition and therapeutics. PMID- 9739493 TI - Genetics and molecular biology. PMID- 9739494 TI - Lipid metabolism. PMID- 9739495 TI - Hyperlipidaemia and cardiovascular disease. PMID- 9739496 TI - Atherosclerosis: cell biology and lipoproteins. PMID- 9739497 TI - Therapy and clinical trials. PMID- 9739498 TI - Is aging a preventable or curable disease? AB - The question of whether aging is a disease is old and controverted. Three possible positions are outlined: (i) aging is a natural event, not a disease; (ii) aging is a disease, to be combated by medical knowledge and skills; and (iii) aging, while natural, can be treated as if it is a disease and efforts made to lessen its undesirable impact. The last position seems, de facto, the one that is being pursued by contemporary medicine; however, in such a pursuit, important issues will be raised of intergenerational justice in paying for advances in medicine, and the need to balance the medical goals of care for aged people against other important social needs. PMID- 9739499 TI - Is the use of some calcium antagonists linked to cancer? Evidence from recent observational studies. AB - In animal and in vitro studies, several calcium antagonists have been shown to block apoptosis (programmed cell death), a natural cellular defence against cancer. On the basis of these studies, it has been hypothesised that calcium antagonists may function as cancer promoters and that they might cause cancer in humans. The association between the use of calcium antagonists and cancer has been addressed recently in 6 independent epidemiological studies. Four of these studies--2 cohort and 2 case-control--found a significantly higher risk of cancer among users of certain calcium antagonists as compared with either non-users or with users of other antihypertensive agents. The other 2 studies failed to find support for this association. All studies had limitations of varying types and significance. The emerging pattern from these investigations includes the following features: (i) the strength of the association appears to be dependent on daily dosage, ranging from no association in users of low dosages to a 2-fold (or possibly higher) increased risk in users of higher dosages; (ii) the time lag to the appearance of this association appears to be at least 2 to 3 years; (iii) an association with a higher risk of cancer has been found primarily for verapamil, while no such relationship has been reported for diltiazem; and (iv) no highly consistent associations have been shown with specific cancer sites or histological types. More data, preferably from long-term randomised clinical trials, are required before firm conclusions can be drawn. PMID- 9739500 TI - Epidemiology and optimal management of polymyalgia rheumatica. AB - Polymyalgia rheumatica (PMR) is a disease of unknown aetiology that occurs in elderly patients, predominantly affecting the Caucasian population. The disease has a slightly higher prevalence in women than in men. There is ongoing discussion regarding the relationship between PMR and giant cell arteritis; an increasing number of studies indicate that they are closely related. PMR has also been linked with rheumatoid arthritis, myopathy and malignant disease. Oral corticosteroids remain the mainstay of drug therapy for PMR. These drugs usually induce prompt relief of symptoms, and some authors consider this dramatic response to be diagnostic for PMR. However, the ideal initial dosage, the duration of treatment and the optimal tapering schedule are much debated. Other drugs, such as methotrexate and azathioprine, have been suggested as corticosteroid sparing agents. Nonsteroidal anti-inflammatory drugs are generally considered to be unsuitable for the long term treatment of PMR. PMID- 9739501 TI - Delirium in the elderly. Optimal management. AB - Delirium is common, morbid and costly, especially among hospitalised elderly patients. Nonetheless, it remains under-recognised and often poorly managed. This article summarises the 5 key steps in the optimal management of delirium. The first step is to precisely define the syndrome of delirium, using key features described in the Diagnostic and Statistical Manual of Mental Disorders (fourth edition) [DSM-IV] or the Confusion Assessment Method. Key features include an acute onset of mental status change, fluctuating course, the presence of inattention, and either disorganised thinking or an altered level of of consciousness. The second step involves the identification of patients at high risk of delirium before it develops, so that preventive measures can be implemented. Risk factors for delirium include advanced age, dementia, impaired functional status, chronic comorbidities and medications, and the severity of the acute illness or surgery. The third step is improved recognition of delirium. Very often, the presence of delirium is neither diagnosed nor properly documented in the medical record. The fourth step is to appropriately evaluate the delirious patient to assess all important contributors to the syndrome. This evaluation will usually involve a careful history, medication review, physical examination and selected laboratory testing. The fifth, and most important, step is the management of the delirious patient. The key elements of management are treating the primary condition(s) leading to delirium, removing all treatable contributing factors, maintaining behavioural control, and supporting the patient and their family. PMID- 9739502 TI - Potential of alpha-glucosidase inhibitors in elderly patients with diabetes mellitus and impaired glucose tolerance. AB - The prevalence of diabetes mellitus (DM) among the elderly, who constitute > 20% of the population in developed countries, is high (up to 40%). Indeed, elderly people represent the bulk (approximately 50%) of the diabetic population. There is much evidence that better glycaemic control can reduce the morbidity associated with this disease. alpha-Glucosidase inhibitors are well tolerated in the treatment of DM in this population. They reduce postprandial hyperglycaemia and have a moderate effect on fastign plasma glucose levels, resulting in a significant reduction in glycated haemoglobin (HbA1C) levels. alpha-Glucosidase inhibitors can be used either as monotherapy or in combination with other oral hypoglycaemic agents or insulin. The good safety profile of these drugs makes them suitable for use in elderly patients with type 2 (non-insulin-dependent) DM, because they can achieve substantial metabolic improvements without any additional risks. Thus, the use of alpha-glucosidase inhibitors should be considered: (i) as a first-choice treatment in newly diagnosed patients; (ii) in individuals whose DM is not well controlled with any other type of treatment; (iii) as an alternative to sulphonylureas or biguanides in patients at risk from hypoglycaemia or lactic acidosis, respectively. Despite the numerous potential advantages of alpha-glucosidase inhibitors in elderly patients with type 2 DM, there is a lack of studies focusing specifically on that population. However, such studies are under way. In addition, the potential of alpha-glucosidase inhibitors in the prevention of type 2 DM and/or on macrovascular disease is currently under study. PMID- 9739503 TI - Rational antithrombotic therapy and prophylaxis in elderly, immobile patients. AB - The aging process is associated with increased coagulation and fibrinolysis parameters, resulting in an overall 'prethrombotic state'. This probably explains the increased baseline susceptibility of elderly patients to the development of thromboembolic disease. Additional factors such as major surgery or malignant disease multiply the risk of thromboembolism in this population. Even when adequate antithrombotic therapy is instituted, the mortality associated with thromboembolic disease remains considerable; this underlines the importance of adequate thromboembolic prophylaxis. At present, the use of low molecular weight heparins (LMWHs) in elderly immobile patients appears to be the most effective approach to prophylaxis. The use of compression stockings seems to be effective in the prevention of venous thrombosis, at least in moderate risk surgical patients. In patients undergoing orthopaedic surgery, additional prophylaxis (e.g. with an LMWH) is necessary. In the management of venous thrombosis, patients can initially be treated with a bodyweight-adjusted dosage of an LMWH. In patients with deep vein leg thrombosis or pulmonary embolism, oral anticoagulant therapy should be started as soon as possible, and should be continued for 6 months. However, before starting prophylaxis or therapy, an individual risk assessment should be performed in which the benefits and disadvantages are balanced. Most of the large trials that have studied the effects of thromboembolic prophylaxis have focused on postsurgical patients. However, it will be of great interest to develop more specific prophylactic and therapeutic regimens for different nonsurgical high risk subgroups of patients, particularly the elderly. PMID- 9739505 TI - Repaglinide. AB - Repaglinide is a novel insulin secretagogue being developed for the management of type 2 (non-insulin-dependent) diabetes mellitus. It stimulates release of insulin from the pancreatic beta-cell, but appears to bind to a different receptor site from sulphonylureas. Repaglinide lowers fasting and postprandial blood glucose levels in animals, healthy volunteers and patients with type 2 diabetes mellitus. Repaglinide is rapidly absorbed and eliminated, which may allow a relatively fast onset and offset of action. Excretion occurs almost entirely by non-renal mechanisms. In comparative clinical trials in patients with type 2 diabetes mellitus, repaglinide 0.5 to 4 mg twice or 3 times daily before meals provided similar glycaemic control to glibenclamide (glyburide) 2.5 to 15 mg/day. Addition of repaglinide to existing metformin therapy resulted in improved glycaemic control. In contrast with glibenclamide, use of repaglinide allowed patients to miss a meal without apparently increasing the risk of hypoglycaemia. PMID- 9739504 TI - Age-associated alterations in calcium current and its modulation in cardiac myocytes. AB - The calcium current is one of the most important components in cardiac excitation contraction coupling. During aging, the magnitude of L-type Ca++ channel current (ICa,L) is significantly increased in parallel with the enlargement of cardiac myocytes, resulting in unaltered ICa,L density. Since the inactivation of ICa,L is slowed and the action potential duration is prolonged, the net Ca++ influx during each action potential is likely to be increased in senescent hearts relative to young ones. This augmentation of Ca++ influx may be important for the preserved cardiac function of the older heart in the basal state. However, it increases the risk of Ca++ overload and Ca(++)-dependent arrhythmias in the senescent heart. During stress, the response of ICa,L to beta-adrenergic receptor stimulation is markedly reduced, which may be an important cause of the age related decrease in cardiac reserve function. These age-dependent changes in ICa,L and its modulations are similar to those observed in the enlarged myocytes of the hypertrophied and failing heart. PMID- 9739506 TI - Gabapentin for disruptive behaviour in an elderly demented patient. PMID- 9739507 TI - Developing a CQI program in a family medicine department. AB - BACKGROUND: Efforts to implement continuous quality improvement (CQI) principles in ambulatory or primary care settings still lag behind efforts in the hospital setting. Many physicians view the concept of CQI with unconcealed skepticism; the process of ambulatory care is very different from that of hospital-based care; and the data necessary to guide CQI efforts are often either missing or inaccurate in the outpatient setting. Since fall 1995, the Department of Family Medicine (DFM) at the University of Michigan (Ann Arbor), including approximately 35 faculty members at seven family practice sites, has been engaged in CQI projects. PLANNING AND IMPLEMENTATION: The CQI committee had a six-month deadline to lay out a plan for educating all faculty and staff in the importance of the CQI approach to problems; design methods for all faculty and staff to buy in to the concepts; and develop a plan to address basic clinical CQI activities, administrative systems change and work environment improvement, and larger ad hoc projects in clinical care, educational programs, and research programs. IMPLEMENTATION: CQI activities were incorporated into the routine monthly business agendas at each clinical site, each of which had a functioning local committee and had begun development of at least one CQI project. PROJECTING INTO THE FUTURE AND CONCLUSIONS: Cost cutting has further moved CQI from the sideline to center stage in the DFM's activities. An effective CQI program can be a major asset in the current competitive health care market, but designing and implementing an outpatient CQI program is a difficult and complex process. Three major problems--the ongoing resistance to change, the slow pace of adding CQI projects to already overburdened work schedules, and the need to conduct the program with ever-decreasing resources available-persist. PMID- 9739508 TI - Measuring the quality of performance in the management of waiting lists: using cataract surgery as an example. AB - BACKGROUND: Quality of care committees monitor waiting lists to ensure that patient care is not compromised. Frequently, waiting lists are determined by individual physicians, and no explicit criteria determine who is first in the queue. The quality of ophthalmologists' decisions for managing waiting lists of cataract patients, a high-volume elective patient group, was examined in a study of patients undergoing cataract surgery in 1997 in the Regina Health District, Saskatchewan, Canada. METHOD: Ninety-eight patients scheduled for surgery were interviewed pre- and postoperatively regarding cataract symptomatology, visual and emotional functioning, coping strategies, and concerns with waiting periods. Ophthalmologists provided preoperative and postoperative information on visual functioning. RESULTS: Even though no formal criteria guided decision making about how long patients should wait, wait periods conformed to general standards set by consensus of ophthalmologists unless patients decided to delay surgery. Patients voiced little concern about the waiting period, and difficulties with visual and emotional functioning were minimal. Surgery outcomes were not negatively affected by waiting periods, which were in part a function of physician case load but were also related to patient preference and the tendency to seek out reassurance. Visual acuity, cataract symptomatology, and visual functioning were not predictive of waiting time, suggesting that this information is not consistently being used to prioritize patients. CONCLUSION: Waiting lists can be well managed by using individual physician decision making, although explicit formal decision making rules would be helpful. A variety of methodologies and analyses can be used to evaluate the management of waiting lists and to assist in identifying criteria for assigning priority to patients. PMID- 9739509 TI - Must patients wait? PMID- 9739510 TI - Identifying and addressing sentinel events: an interview with Richard Croteau. Interview by Steven Berman. PMID- 9739511 TI - [Measurement of coronary flow reserve using adenosine 5'-triphosphate in dogs]. AB - Adenosine 5'-triphospate (ATP) was compared with adenosine and papaverine for the measurement of coronary flow reserve in 12 anesthetized dogs. Intracoronary bolus injection of ATP (1 ml, 1-500 microM) produced a dose dependent increase in the blood flow of the left anterior descending artery, which attained the plateau at the dose of 100 microM. The ratio of peak to resting coronary flow volume (coronary flow reserve) with 100 microM of ATP (3.5 +/- 0.5) was similar to that with 200 microM of adenosine (4.0 +/- 0.7) and 50 mM of papaverine (3.7 +/- 0.8). Hemodynamic variables did not change after administration of each drug, except left ventricular regional wall motion abnormality during papaverine injection. The coronary flow reserve as measured after intracoronary ATP administration (100 microM) decreased as the grade of stenosis of the left anterior descending artery progressed. In addition, the flow reserve was similar to that of adenosine or papaverine administration at each stenosis grade. Intravenous administration of ATP (1,000 micrograms/min) caused a similar increase in coronary blood flow as intracoronary ATP injection (100 microM). However, premedication with 8 phenyltheophylline, an adenosine receptor blocker, significantly suppressed the coronary dilatory effect of intravenous ATP and intracoronary adenosine but not the effect of intracoronary ATP. These results indicate that intracoronary ATP is useful for measuring coronary flow reserve and that its coronary dilatory effect is not mediated by metabolysis to adenosine. PMID- 9739512 TI - [Experience of coronary and great vessel angiography by transradial puncture]. AB - The introduction of 5F and even 4F catheters allows transradial coronary arteriography and aortography. The efficacy and limitation of angiography via the radial artery using 5F catheter was prospectively evaluated in 200 consecutive patients. Cardiac catheterization with diagnostic angiography was successfully performed in 198 of 200 patients, including 11 patients with acetylcholine provocation test, 21 with bypass graft angiography, 38 with aortography and 5 with biopsy of the left ventricular myocardium. The transradial approach was not indicated in one patient without normal Allen's test and in one with weak radial pulse. In four patients, guide wire support was needed during manipulation because of marked tortuosity in the innominate artery. The sheath was removed immediately after the completion of the procedure, followed by 5 hours of tourniquet hemostasis without manual compression. The postoperative resting period was reduced. Peripheral vasospasm occurred in 2.5% of cases, but could be eliminated by administration of isosorbide dinitrate and lidocaine. Subcutaneous hemorrhage in the puncture site was observed in 3.0% of cases, but required no additional compression. Transradial catheterization is a minimally invasive, safe and practical alternative to the brachial or femoral artery approach in patients with normal Allen's test. PMID- 9739513 TI - [Echocardiographic and hematological variables as a risk factor for stroke in chronic nonvalvular atrial fibrillation]. AB - The relationship between echocardiographic variables and the incidence of ischemic stroke in patients with atrial fibrillation was investigated by transthoracic and transesophageal echocardiography in 67 patients with chronic nonvalvular atrial fibrillation. Hematologic variables were also measured simultaneously, including plasma levels of D-dimer and thrombin-antithrombin III complex in these patients. There was a prior history of ischemic stroke in 13 patients (stroke group), but not in the other 54 patients (nonstroke group). There were no significant differences in age, sex, left ventricular ejection fraction, left ventricular end-diastolic diameter, left atrial diameter or hematologic parameters between the groups. The left atrial appendage emptying flow velocity was lower in the stroke group than in the nonstroke group (21 +/- 5 vs 32 +/- 3 cm/sec, p < 0.05), and the incidence of left atrial spontaneous echo contrast was significantly higher in the stroke group than in the nonstroke group (69% vs 26%, p < 0.01). There was no significant difference in the incidence of left atrial thrombi between the groups (23% vs 12%). These findings suggest that transesophageal echocardiographic variables are correlated with the risk of ischemic stroke in patients with chronic nonvalvular atrial fibrillation. PMID- 9739514 TI - [Evaluation of patients with cerebral infarction using transesophageal echocardiography: atherosclerotic changes in the thoracic aorta and the branches of the aortic arch]. AB - Atherosclerotic disease of the aortic arch is thought to be a potential source of cerebral emboli, but this disease in the branch of the aortic arch has not been extensively explored. This study assessed atherosclerotic lesions in the thoracic aorta and the branches of the aortic arch using transesophageal echocardiography in patients with cerebral infarction, and simultaneously searched for potential cardiac sources for emboli. Thrombi were detected in the left atrial appendage in nine of 54 patients with cerebral infarction and these patients were excluded. The remaining 45 patients with cerebral infarction (31 males and 14 females aged 68.5 +/- 7.4 years) and 35 normal subjects (21 males and 14 females aged 69.2 +/- 9.5 years) were evaluated. The thickness of the wall was measured in the branches of the aortic arch (brachiocephalic trunk, left common carotid artery and left subclavian artery) as well as the thoracic aorta (ascending aorta, aortic arch and descending aorta). Atherosclerotic lesions were defined as increased echogenicity of the intima (intimal thickening), calcification, protruded plaque, ulceration or plaque with cystic lesion. The thicknesses of the wall in the aortic arch (3.84 +/- 1.25 vs 2.71 +/- 1.33 mm, p < 0.01), left common carotid artery (2.67 +/- 1.10 vs 2.16 +/- 0.91 mm, p < 0.05) and the left subclavian artery (2.52 +/- 0.67 vs 2.15 +/- 0.88 mm, p < 0.05) were significantly greater in patients than in the normal subjects. The incidence of plaque or ulceration was significantly increased in patients with cerebral infarction compared with the normal subjects in the aortic arch (76% vs 43%, p < 0.05) and left common carotid artery (44% vs 17%, p < 0.05). Transesophageal echocardiography can detect possible sources of emboli in the branches of the aortic arch as well as the thoracic arch in patients with cerebral infarction. PMID- 9739516 TI - [A 26-year-old man complaining of low grade fever]. PMID- 9739515 TI - [Left atrial function and left atrial appendage flow velocity in hypertrophic cardiomyopathy: comparison of patients with and without paroxysmal atrial fibrillation]. AB - The involvement of left atrial (LA) appendage flow velocity in reduced left atrial function was investigated in 24 patients with hypertrophic cardiomyopathy, who retained sinus rhythm at the examination. Patients were divided into 11 with a history of paroxysmal atrial fibrillation [PAf(+)] and 13 without such history [PAf(-)]. Transthoracic echocardiography was performed to evaluate LA fractional shortening (LA%FS) and mean velocity of circumferential LA fiber shortening (LAmVcf), as contractile functions of the left atrium at the phase of active atrial contraction. Transesophageal echocardiographic Doppler examination was performed in all patients to measure the LA appendage velocity. In all patients, significant positive correlations were observed between the LA appendage velocity and LA%FS (r = 0.50, p < 0.05) or LAmVcf (r = 0.82, p < 0.001). LAmVcf and LA appendage velocity in patients with paroxysmal fibrillation were significantly lower than in those without (0.84 +/- 0.15 vs 1.28 +/- 0.37 circ/sec, 44 +/- 12 vs 65 +/- 20 cm/sec, both p < 0.01), whereas LA diameter was greater in the former compared to the latter (45 +/- 5 vs 38 +/- 5 mm, p < 0.01). LAmVcf and LA appendage velocity were low in four patients with cerebral infarction or transient cerebral ischemic attack (LAmVcf < 1.0 circ/sec, LA appendage velocity < or = 40 cm/sec). Importantly, all these patients had a history of paroxysmal fibrillation. These results indicate that there is a close relationship between LA appendage velocity and LA contractile function in patients with hypertrophic cardiomyopathy with paroxysmal atrial fibrilation, and these patients have potential risk of cerebral infarction. PMID- 9739517 TI - [Glutamate antagonist]. PMID- 9739518 TI - [Insulin-like growth factor I]. PMID- 9739519 TI - [Treatment with lecithinized superoxide dismutase in amyotrophic lateral sclerosis]. PMID- 9739520 TI - [Neurotrophic factor treatment in amyotrophic lateral sclerosis]. PMID- 9739521 TI - [Effect of somatosensory disturbance to outcomes of motor function in thalamic hemorrhage: evaluation with short-latency somatosensory evoked potential (SSEP)]. AB - We investigated the effect of the somatosensory functions to the outcomes of motor functions in 28 patients with thalamic hemorrhage. The disturbance of the pyramidal tracts was assessed by the destruction of the internal capsule found in computed tomography (CT). The disturbance of the somatosensory function was analyzed by the N20 component of short-latency somatosensory evoked potentials (SSEP). The outcomes of motor function was evaluated after 3 months of ictus. Correlations between the outcomes of motor function, disturbance of the pyramidal tract, and disturbance of the somatosensory function were discussed. The result indicated that functional outcomes statistically correlated with neither disturbance of the internal capsule alone nor disturbance of N20 alone. But, there was statistically significant between functional outcomes and the combination of disturbance of the internal capsule with disturbance of N20 (p < 0.05, Wilcoxon signed-rank). There was not statistical difference in hematoma volume or consciousness. The implications of these results suggest that somatosensory function may affect the recovery of motor functions. PMID- 9739522 TI - [Histological and morphometrical studies on small arteries in normal brainstems with special reference to the pathogenesis of the hypertensive brainstem hemorrhage]. AB - Plasmatic arterial necrosis and microaneurysm of small arteries are preceded by smooth muscle cell loss and the rupture of these arterial lesions is a direct cause of hypertensive intracerebral hemorrhage. The hypertensive brainstem hemorrhage occur exclusively in the pons. To elucidate whether there are differences of underlying arterial lesions between each part of the brainstem or not, small arteries in normal 34 autopsied brainstems were investigated histologically and morphometrically. Histological study revealed the predilection of the occurrence of plasmatic arterial necrosis, microaneurysms and fibronodular arterial lesions in the hypertensive pons. These lesions occurred predominantly in the small arteries 100-300 microns in diameter in the basal part of hypertensive pons, and were rare in the other parts of hypertensive brainstems and in normotensive brainstems. A negative correlation between the ratio of number of smooth muscle cell nuclei to the area of tunica media and age was demonstrated morphometrically. The ratio in the hypertensive group was significantly lower than that of the normotensive group. In addition the mean ratio in the pons was significantly lower than that in the midbrain and the medulla oblongata in the hypertensive group. These results are consistent with the fact that the hypertensive brainstem hemorrhage predominantly occur in the pons and primary bleedings in the other parts of the brainstem are rare. PMID- 9739523 TI - [The effect of hippocampal dentate granule cell lesions upon the limbic seizure model of rats]. AB - The purpose of this study was to determine the role of dentate granule cells upon limbic seizure of Wistar rat caused by unilateral intra-amygdaloid administration of kainic acid (KA). Stereotactic surgery was performed in Wistar rats and stainless steel injection chemitorode was inserted in the left amygdala. Left dentate granule cells lesion were induced by microinjection of colchicine. The rats obtained recovery period for 7 days, postoperatively. The rats were divided into two groups. One group were used for observation of symptoms and electroencephalographic findings during the limbic seizure for 6 hours after the KA injection. Another group was processed for measuring local cerebral glucose utilization (LCGU) during limbic seizure status. The histological study demonstrated a selective loss of dentate granule cells in the left hippocampus 7 days after the colchicine injection. After the KA injection, initiation of the spike discharge was significantly retarded not only in the hippocampus (from 6.01 min. to 37.25 min.) but also in the amygdala (from 2.96 min. to 10.8 min.). Progression, frequency and intensity of the KA induced seizures were also inhibited by the colchicine-induced dentate granule cells lesion. During limbic seizure status, LCGU obtained by 14C-deoxyglucose autoradiography were significantly decreased not only in the hippocampus but also in the amygdala on the site of KA injection. These data suggest that hippocampal dentate granule cells play an important role on initiation and progression of the KA induced limbic seizure. The result suggested that there was an acceleration mechanism of the limbic seizure between amygdala and hippocampus. PMID- 9739524 TI - [Chronic progressive radiation myelopathy after bone marrow transplantation]. AB - Two patients after bone marrow transplantation (BMT) developed chronic progressive radiation myelopathy (CPRM). The factors contributing to development of CPRM at the low dose, which radiation doses given for enlarged regional lymph nodes prior to BMT ordinarily would be too low to induce CPRM, were discussed, and clinicoradiologic correlations in CPRM from onset through the stabilized state examined. The clinicoradiologic findings of two patients, who are a 26-year old man with malignant lymphoma (autologous BMT) performed radiotherapy totaling 20 Gy for enlarged regional lymph nodes before BMT, and a 40-year old woman with chronic myeloid leukemia (allogenic BMT) done 18.9 Gy, were examined. The involved spinal cord segments were irradiated for lymph node enlargement prior to BMT, and all the clinicoradiologic findings were consistent with CPRM. We considered possible synergistic toxicity with high-dose busulfan accompanying BMT. Unlike the second case, the first patient had continued severe progression of CPRM, possibly because a higher dose of additional radiotherapy (30 Gy) was given for presumed spinal cord tumor involvement in that case than in the other (< 20 Gy). These cases demonstrate that BMT protocols carry a risk of potentiating the spinal cord toxicity of low-dose radiotherapy. PMID- 9739525 TI - [Dissecting aneurysm of the extracranial vertebral artery causing TIA: a case report]. AB - Spontaneous dissecting aneurysm of the extracranial vertebral artery is not frequent. We report a case of dissecting aneurysm of the left extracranial vertebral artery causing TIA. A 44-year-old man was admitted to our hospital with severe occipitalgia of sudden onset on Dec. 21, 1995. On admission, he presented with left hemiparesis and left hemidysesthesia. CT scan and SPECT showed no abnormal findings. His neurological deficits completely improved on the following day. Initial cerebral angiography performed on Dec. 27, 1995 showed pearl and string sign at V3 segment and irregular stenosis at V2 segment suggesting a dissecting aneurysm. We treated conservatively. Repeated angiography performed four weeks after the onset showed resolution of the stenosic lesion and disappearance of the aneurysm. We discussed diagnosis and etiology and treatment of dissecting aneurysm of the extracranial vertebral artery. PMID- 9739526 TI - [Peripheral facial palsy]. PMID- 9739527 TI - [Jugular foramen meningioma]. PMID- 9739528 TI - [A 74-year-old woman with parkinsonism and dementia who died four years after the onset]. AB - We report a 74-year-old woman with parkinsonism and dementia, who died 4 years after the onset of the disease. She was well until 70 years of the age (1993) when she noted slowness in the movement in her left hand. She also developed gait disturbance and the similar symptoms spread to the right upper and lower extremities. Two years after the onset, she had difficulty in walk, and was admitted to our hospital on March 9, 1995. Her daughter had the onset of hand tremor at 50 years of the age and gait disturbance at 52. Her gait improved after levodopa treatment, but her MRI revealed a liner T2-high signal lesion along the outer surface of each putamen. On admission, the patient was alert but slighted demented. Higher cerebral functions were normal. She had a masked face and small voice. Her gait was of small step without arm swing. Retropulsion was present. Rigidity was noted in the neck but not in the limbs. She was bradykinetic but tremor was absent. She was treated with levodopa/carbidopa, dops, and bromocriptine with considerable improvement and was discharged on March 30, 1995. On January 19, 1996, she developed fever and hallucination; she became more akinetic and admitted again. She showed marked dementia and stage IV parkinsonism. She was treated by supportive measures with improvement in the general condition, but she was found to have a gastric cancer for which a subtotal gastrectomy was performed on March 11, 1996. Post-operative course was uneventful, but her parkinsonism progressed to stage V. She was transferred to another hospital on May 13, 1996. In July 21, 1996, she developed dyspnea and fever and was admitted to our hospital again. She was somnolent. Rigidity was moderate to marked and she was unable to stand or walk. By supportive cares, her general condition improved and was discharged to home on November 4, 1996. She developed fever on June 13, 1997 and admitted to our service again. Her BP was 150/90 mmHg. She was alert but markedly demented. Laboratory examination revealed increases in liver enzymes (GOT 75 IU/l, GPT 101 IU/l) and renal dysfunction (BUN 68 mg/dl, creatinine 3.27 mg/dl). Subsequent hospital course was complicated by renal failure and thrombocytopenia (33,000/ml). She expired on July 1, 1997. The patient was discussed in a neurologic CPC, and a chief discussant arrived at the conclusion that the patient had diffuse Lewy body disease and her daughter striatonigral degeneration. Some participants thought both the patient and her daughter had diffuse Lewy body disease. Post-mortem examination revealed marked degeneration of the substania nigra and the locus coeruleus. The medial part of the nigra also showed marked cell loss. Lewy bodies were found in the remaining nigral and coeruleus neurons. Cortical Lewy bodies were very few and the striatum was intact. Pathologic diagnosis was Parkinson's disease. Dementia was in part attributed to the marked degeneration of the medial part of the substantia nigra. PMID- 9739529 TI - The chance of a lifetime--nursing's golden opportunity. PMID- 9739530 TI - Medicare payment for advanced practice nurses: what are the research questions? PMID- 9739531 TI - Is it over yet? Hope or hype: status of the HIV/AIDS pandemic. PMID- 9739532 TI - Interdisciplinary team: the nursing perspective is essential. PMID- 9739533 TI - Cross-country truck drivers: a vulnerable population. PMID- 9739534 TI - Theory construction based on standards of care: a proposed theory of the peaceful end of life. AB - The contribution of developing a theory from this standard of care is that it can express a new unifying idea about the phenomenon of peaceful end of life for terminally ill patients. It allows for generating and testing hypotheses that can provide new insights into the nature of this phenomenon and can contribute to increased knowledge about nursing interventions that help patients toward a peaceful end of life. The process of theory development from standards of care as described in this article also can be applied to other phenomena. Clinical practice abounds with opportunities for theory development, yet nurses often do not use theories to guide their practice. Until now, little guidance has been provided to tap the richness of clinical knowledge for the development of middle range theories. Whereas the method described in this article may still be further refined, it offers a promising approach for the development of theories that are applicable to practice and move beyond the scope of grand theories. Thus deriving theories from standards of care can offer an important contribution to the development of the discipline's scientific knowledge base and enhanced practice. PMID- 9739535 TI - Caring interactions among nursing students: a descriptive comparison of 2 associate degree nursing programs. AB - Increasing emphasis has been given to the importance of caring as a curricular theme in nursing education and the value of developing students who will be able to function in a practice role as a caring nurse. The challenge for nurse educators is to understand and facilitate those processes through which students can be socialized to caring as a professional value in nursing. The findings from research that was conducted to investigate caring in nursing education suggest that caring can emerge during students' interactions with other students and during their interactions with faculty members. Therefore, it may be useful to capitalize on peer relationships as a strategy through which students can experience caring behaviors and develop skill in the practice of caring for others. The findings from this study support the need to investigate experiential learning strategies that are designed to promote the practice of caring among the student peer group. Additional research is needed to validate the effectiveness of such strategies as a method through which nursing students can learn caring. PMID- 9739536 TI - Increasing the supply of doctoral students. PMID- 9739537 TI - Overtraining and recovery. A conceptual model. AB - Fiercer competition between athletes and a wider knowledge of optimal training regimens dramatically influence current training methods. A single training bout per day was previously considered sufficient, whereas today athletes regularly train twice a day or more. Consequently, the number of athletes who are overtraining and have insufficient rest is increasing. Positive overtraining can be regarded as a natural process when the end result is adaptation and improved performance: the supercompensation principle--which includes the breakdown process (training) followed by the recovery process (rest)--is well known in sports. However, negative overtraining, causing maladaptation and other negative consequences such as staleness, can occur. Physiological, psychological, biochemical and immunological symptoms must be considered, both independently and together, to fully understand the 'staleness' syndrome. However, psychological testing may reveal early-warning signs more readily than the various physiological or immunological markers. The time frame of training and recovery is also important since the consequences of negative overtraining comprise an overtraining-response continuum from short to long term effects. An athlete failing to recover within 72 hours has presumably negatively overtrained and is in an overreached state. For an elite athlete to refrain from training for > 72 hours is extremely undesirable, highlighting the importance of a carefully monitored recovery process. There are many methods used to measure the training process but few with which to match the recovery process against it. One such framework for this is referred to as the total quality recovery (TQR) process. By using a TQR scale, structured around the scale developed for ratings of perceived exertion (RPE), the recovery process can be monitored and matched against the breakdown (training) process (TQR versus RPE). The TQR scale emphasises both the athlete's perception of recovery and the importance of active measures to improve the recovery process. Furthermore, directing attention to psychophysiological cues serves the same purpose as in RPE, i.e. increasing self-awareness. This article reviews and conceptualises the whole overtraining process. In doing so, it (i) aims to differentiate between the types of stress affecting an athlete's performance: (ii) identifies factors influencing an athlete's ability to adapt to physical training: (iii) structures the recovery process. The TQR method to facilitate monitoring of the recovery process is then suggested and a conceptual model that incorporates all of the important parameters for performance gain (adaptation) and loss (maladaptation). PMID- 9739538 TI - Saliva composition and exercise. AB - Little attention has been directed toward identifying the changes which occur in salivary composition in response to exercise. To address this, our article first refers to the main aspects of salivary gland physiology. A knowledge of the neural control of salivary secretion is especially important for the understanding of the effects of exertion on salivary secretion. Both salivary output and composition depend on the activity of the autonomic nervous system and any modification of this activity can be observed indirectly by alternations in the salivary excretion. The effects of physical activity (with reference to factors such as exercise intensity and duration, or type of exercise protocol) on salivary composition are then considered. Exercise might indeed induce changes in several salivary components such as immunoglobulins, hormones, lactate, proteins and electrolytes. Saliva composition might therefore be used as an alternative noninvasive indicator of the response of the different body tissues and systems to physical exertion. In this respect, the response of salivary amylase and salivary electrolytes to incremental levels of exercise is of particular interest. Beyond a certain intensity of exercise, and coinciding with the accumulation of blood lactate (anaerobic threshold or AT), a 'saliva threshold' (Tsa) does indeed exist. Tsa is the point during exercise at which the levels of salivary alpha-amylase and electrolytes (especially Na+) also begin to rise above baseline levels. The occurrence of the 2 thresholds (AT and Tsa) might, in turn, be attributable to the same underlying mechanism, that of increased adrenal sympathetic activity at high exercise intensities. PMID- 9739539 TI - Lipid metabolism during exercise. AB - Fat is an extremely important substrate for muscle contraction, both at rest and during exercise. Triglycerides (TGs), stored in adipose tissue and within muscle fibres, are considered to be the main source of the free fatty acids (FFAs) oxidised during exercise. It is still unclear, however, how the use of these substrates is regulated during exercise. The regulation seems to be multifactorial and includes: (i) dietary and nutritional status; (ii) hormonal milieu; (iii) exercise mode, intensity and duration; and (iv) training status. On the other hand, the mechanism for FFA transport from its storage as triglycerides in adipose tissue and muscle to its place of utilisation in heart, skeletal muscle, kidney and liver is more clearly understood. It has been determined that the plasma FFA turnover rate is sufficiently rapid to account for most of the fat metabolised during low intensity exercise (25 to 40% VO2max). However, an exercise intensity of 65% VO2max results in a slight decrease in the amount of plasma FFA uptake by muscle tissue. Other studies have found that during prolonged exercise, muscle TGs become the predominant source of energy obtained from fat. Furthermore, it is widely documented that endurance activities increase the energy utilisation from fat while sparing carbohydrate sources. For example, during exercise on a cycle ergometer, nonplasma FFAs and plasma FFAs contribute 40%, and carbohydrates 60%, of the total calculated amount of energy expenditure before exercise and vice versa after exercise (60% nonplasma and plasma FFAs and 40% carbohydrates). Although it was many years before it was fully demonstrated, fat is now known to be transported in the blood as FFA bound to the protein carrier albumin. The mobilisation of FFA is primarily a function of sympathetic nervous activity directed towards the adipocytes, or the 'fat pad'. This nervous activity can be direct or may be an effect of circulating catecholamines such as adrenaline (epinephrine). This article summarises the role of fat metabolism during exercise. PMID- 9739540 TI - Golf injuries. An overview. AB - Over the years, golf has become an increasingly popular sport, attracting new players of almost all ages and socioeconomic groups. Golf is practised by up to 10 to 20% of the overall adult population in many countries. Beyond the enjoyment of the sport itself, the health-related benefits of the exercise involved in walking up to 10 km and of relaxing in a pleasant natural environment are often reported to be the main motives for adhering to this activity by recreational golfers. Golf is considered to be a moderate risk activity for sports injury; however, excessive time spent golfing and technical deficiencies lead to overuse injuries. These are the 2 main causes of injuries among golfers, and each has specific differences in the pattern in which they occur in professional and amateur golfers. Golf injuries originate either from overuse or from a traumatic origin and primarily affect the elbow, wrist, shoulder and the dorsolumbar sites. Professional and weekend golfers, although showing a similar overall anatomical distribution of injuries by body segment, tend to present differences in the ranking of injury occurrence by anatomical site; these differences can be explained by their playing habits and the biomechanical characteristics of their golf swing. Many of these injuries can be prevented by a preseason, and year round, sport-specific conditioning programme including: (i) muscular strengthening, flexibility and aerobic exercise components; (ii) a short, practical, pre-game warm-up routine; and (iii) the adjustment of an individual's golf swing to meet their physical capacities and limitations through properly supervised golf lessons. Finally, the correct selection of golf equipment and an awareness of the environmental conditions and etiquette of golf can also contribute to making golf a safe and enjoyable lifetime activity. PMID- 9739541 TI - Prevention of ankle injuries. PMID- 9739542 TI - 9-Hydroxy-9-(3-methylbut-3-en-1-ynyl)-9H-fluorene-1-carbonitrile. AB - The N atom in the title compound, C19H13NO, lies -0.0365 (14) A from the plane formed by the fluorene system, and the C [symbol: see text] N distance is 1.143 (2) A. The C [symbol: see text] C distance is 1.191 (2) A, and the ethynyl group deviates slightly from linearity, with C-C [symbol: see text] C-C bond angles of 172.0 (1) and 176.1 (2) degrees. Terminal CH2 and CH3 groups are 50:50 disordered, with equal C-C distances of 1.408 (3) and 1.406 (3) A. PMID- 9739543 TI - Phenanthrene-4-carboxylic acid and 1,2-dihydrophenanthrene-4-carboxylic acid. AB - Phenanthrene-4-carboxylic acid, C15H10O2, crystallized in the centrosymmetric space group P2(1)/n, while 1,2-dihydrophenanthrene-4-carboxylic acid, C15H12O2, crystallized in the centrosymmetric space group Pbca. In each structure, there is a single type of hydrogen bond: it is of the cyclic dimer type about a center of symmetry. The Odonor...Oacceptor distances are 2.634 (2) and 2.651 (2) A, and the O-H...O angles are 176 (3) and 173 (2) degrees, respectively, for the two structures. In each structure, the carboxy H and O atoms are ordered. The phenanthrene core of the fully aromatic acid is roughly planar; the dihedral angle between the best-fit core plane and the carboxy group plane is 63.7 (1) degree. As expected, the hydrogenated ring of the second acid is much less nearly planar; the remaining naphthalenoid core is, however, roughly planar and the dihedral angle between this best-fit plane and the carboxy group plane is 60.4 (1) degree. PMID- 9739544 TI - 8-Aminocaprylic acid. AB - The title acid, 8-aminooctanoic acid, C8H17NO2, crystallized in the centrosymmetric space group P2(1)/n in the zwitterionic form. The three H atoms involved in hydrogen bonding are ordered. The five intermolecular N-H...O hydrogen bonds have N...O distances ranging from 2.752 (2) to 3.258 (2) A and N H...O angles ranging from 131 (2) to 165 (2) degrees. Each molecule is linked to six neighboring molecules by a total of ten hydrogen bonds. A complex network of hydrogen bonds ensues in which chains predominate. PMID- 9739545 TI - DL-3-aminoisobutyric acid monohydrate. AB - The title acid, 3-amino-2-methylpropanoic acid monohydrate, C4H9NO2.H2O, crystallized in the centrosymmetric space group Pbca in the zwitterionic form. The three H atoms on N, which are involved in hydrogen bonding, are ordered. The three intermolecular N-H...O hydrogen bonds have N...O distances ranging from 2.758 (2) to 2.809 (2) A and N-H...O angles ranging from 149 (2) to 171 (1) degrees. The two intermolecular O-H...O hydrogen bonds have O...O distances 2.739 (2) and 2.755 (2) A, and O-H...O angles 170 (2) and 175 (2) degrees. Each acid molecule and its associated water molecule are directly hydrogen bonded to five acid molecules and two water molecules; the structure comprises two subsets of molecules which are not cross-linked by these hydrogen bonds. Through basic second-level graphs, approximately two-thirds of the hydrogen-bonding patterns are finite and one-third are chains; there is a single ring pattern, which occurs about a center of symmetry. PMID- 9739546 TI - Choosing objective lenses: the importance of numerical aperture and magnification in digital optical microscopy. AB - Microscopic images are characterized by a number of microscope-specific parameters--numerical aperture (NA), magnification (M), and resolution (R)--and by parameters that also depend on the specimen--for example, contrast, signal-to noise ratio, dynamic range, and integration time. In this article, issues associated with the microscope-specific parameters NA, M, and R are discussed with respect to both widefield and laser scanning confocal microscopies. Although most of the discussion points apply to optical microscopy in general, the main application considered is fluorescence microscopy. PMID- 9739547 TI - Sickling of anoxic red blood cells in fish. AB - The occurrence of the mutant hemoglobin Hb S in human red blood cells results in sickle cell anemia. This disease, including its genetic and molecular bases, has been extensively investigated and is well understood (1,2). The presence of deoxy induced sickling of animal erythrocytes is largely unknown, however. We examined red blood cells (RBCs) from several fish species in vitro under aerated and anoxic conditions. Our polarized light microscopic techniques were aimed at establishing correlations between erythrocyte morphology, state of oxygenation, spectral absorbance, linear dichroism, and linear birefringence. We found no fish with intracellular HbO2 polymerization; but there were intraerythrocytic aggregations of deoxy Hb with a high degree of either molecular order or disorder. The ordered aggregates in the RBCs of Atlantic cod, haddock, and toadfish were remarkably similar in dichroic ratio magnitudes and birefrigence to those in human RBCs that contain HbS. Therefore, fish hemoglobins appear to be good models of sickling disorders and polymerization-related phenomena. The consequences of sickling on animal health and fish aquaculture remain to be studied. PMID- 9739548 TI - Calcium speciation and exchange between blood and extrapallial fluid of the quahog Mercenaria mercenaria (L.). AB - Calcium and small organic molecules (e.g., tyrosine, MW 181 Da) introduced into the extrapallial fluid (EPF) of the quahog Mercenaria mercenaria exhibit rapid fluxes across the outer mantle epithelium and are distributed throughout the circulatory system within 3 h. Larger molecules (e.g., bovine serum albumin, MW 66,000 Da) are less readily exchanged between EPF and blood. The protein compositions of blood plasma and EPF are different, with at least seven protein bands expressed more prominently in the EPF. Equilibrium dialysis experiments reveal that Ca2+ constitutes only 2% of the total Ca in plasma; most of the Ca (85%) is bound to macromolecules, and the remaining 13% is present as dialyzable low molecular weight moieties. This distribution cannot be explained by speciation of inorganic Ca alone, since the MINTEQA2 equilibrium speciation model predicts that 79%-86% of the Ca should be present as Ca2+, with the remainder as CaSO4 (20%-13%). However, inclusion of a weakly Ca-binding organic molecule (log10 Ka approximately 2 M-1) into MINTEQA2 could fully reconcile modeling with experimental measurements. Results suggest that calcium transport in blood plasma and EPF is mediated by a suite of proteins and small organic ligands with a low affinity for Ca. PMID- 9739549 TI - Isolation and characterization of endostyle-specific genes in the ascidian Ciona intestinalis. AB - The endostyle is a special organ in the pharynx of Urochordata, Cephalochordata, and Cyclostomata. It may have arisen in the common ancestor of these taxa, along with a shift to internal feeding for extracting suspended food from the water. In addition, the endostyle has a functional homology to the vertebrate thyroid gland. The endostyle is therefore one of the structures key to the understanding of the origin and evolution of chordates. In the present study, we isolated and characterized cDNA clones for four endostyle-specific genes, CiEnds1, CiEnds2, CiEnds3, and CiEnds4, of the ascidian Ciona intestinalis. Although the predicted amino acid sequences of the gene products CiENDS1, CiENDS2, and CiENDS3 showed no similarity to known proteins, their mean hydropathy profiles suggest that they are secretory proteins. In addition, CiENDS3 contained a unique repeat of 10 amino acids [R(QPCI)-(RRPC)I]. CiEnds1 and CiEnds2 were expressed in zone 6, a protein-secreting glandular element of the endostyle, and CiEnds3 was expressed in zone 2, another secretory zone. CiEnds4, a cytoplasmic actin gene, was predominantly expressed in zones 3 and 5, which are supporting elements of the endostyle. The amino acid sequences of CiENDS1 and CiENDS2 resembled each other. In addition, they resembled a zone-6-specific gene product (HrENDS2) of another ascidian, Halocynthia roretzi. The results suggest that these genes are conserved among ascidian species, and therefore they (as well as CiEnds3 for the protein with a unique motif) may be useful probes for further analyses of molecular mechanisms involved in endostyle development. PMID- 9739550 TI - Molecular phylogeny of zooxanthellate bivalves. AB - The aim of this research was to analyze the phylogenetic relationships of zooxanthellate bivalves belonging to the genera Tridacna, Hippopus, Fragum, and Corculum as well as to the closely related azooxanthellate bivalves belonging to Vasticardium and Fulvia. The small-subunit ribosomal RNA genes (18S rDNAs) from these bivalves were amplified by polymerase chain reaction with universal eukaryotic primers and were then sequenced. The sequence data from each species were analyzed by the neighbor-joining, maximum parsimony, and maximum likelihood methods, and phylogenetic trees were constructed. The results were essentially consistent with the morphological taxonomy of these bivalves. Thus, the zooxanthellate clams branch into two lineages, one composed of the genera Fragum and Corculum in the family Cardiidae, and the other composed of the genera Tridacna and Hippopus in the family Tridacnidae. However, present results indicate that the azooxanthellate clams analyzed (Vasticardium flavum and Fulvia mutica) are more likely to form a clade with the species of Tridacna and Hippopus than with those of Fragum and Corculum. This topology suggests that either the symbiosis with zooxanthellae occurred independently in each of two lineages, Tridacna-Hippopus and Corculum-Fragum, or the symbiosis was established in clams ancestral to the lineages of both the zooxanthellate clams and the azooxanthellate clams Vasticardium and Fulvia, and the latter lost the symbiotic relationship after the symbiotic clam lineages had diverged. PMID- 9739551 TI - Genetic engineering for high methionine grain legumes. AB - Methionine (Met) is the primary limiting essential amino acid in grain legumes. The imbalance in amino acid composition restricts their biological value (BV) to 55 to 75% of that of animal protein. So far improvement of the BV could not be achieved by conventional breeding. Therefore, genetic engineering was employed by several laboratories to resolve the problem. Three strategies have been followed. A) Engineering for increased free Met levels; B) engineering of endogenous storage proteins with increased numbers of Met residues; C) transfer of foreign genes encoding Met-rich proteins, e.g. the Brazil nut 2S albumin (BNA) and its homologue from sunflower, into grain legumes. The latter strategy turned out to be most promising. In all cases the gene was put under the control of a developmentally regulated seed specific promoter and transferred into grain legumes using the bacterial Agrobacterium tumefaciens-system. Integration into and copy numbers in the plant genome as well as Mendelian inheritance and gene dosage effects were verified. After correct precursor processing the mature 2S albumin was intracellularly deposited in protein bodies which are part of the vacuolar compartment. The foreign protein amounted to 5 to 10% of the total seed protein in the best transgenic lines of narbon bean (Vicia narbonensis L., used in the authors' laboratories), lupins (Lupinus angustifolius L., used in CSIRO, Australia), and soybean (Glycine max (L.) Merr., used by Pioneer Hi-Bred, Inc., USA). In the narbon bean the increase of Met was directly related to the amount of 2S albumin in the transgenic seeds, but in soybean it remained below the theoretically expected value. Nevertheless, trangenic soybean reached 100%, whereas narbon bean and lupins reached approximately 80% of the FAO-standard for nutritionally balanced food proteins. These results document that the Met problem of grain legumes can be resolved by genetic engineering. PMID- 9739552 TI - Production of genetically modified lysozymes having extreme heat stability and antimicrobial activity against gram negative bacteria in yeast and in plant. AB - Hen egg white lysozyme was genetically modified to have extreme heat stability and strong antimicrobial activity against Gram negative bacteria and the modified lysozymes were secreted in yeast and tobacco. Complementary DNA encoding lysozyme was subjected to site-directed mutagenesis to have the Asn-X-Thr(Ser) sequence that is the signal for asparagine-linked glycosylation at the positions 49. The glycosyl lysozyme enhanced heat stability was expressed in the yeast carrying the modified lysozyme cDNA. The expression amount of glycosyl lysozyme was about 10 mg/l of yeast culture medium. Using the same yeast expression system, the lysozyme enhanced antimicrobial action by inserting hydrophobic penta-peptide at the C-terminus were secreted in a small amount (less than 100 micrograms/l in the yeast culture medium). These cDNA constructs of modified lysozymes were engineered into tabacco through Agrobacterium-mediated transformation in order to construct antimicrobial plant. The expression of lysozymes was confirmed by the reverse transcriptional PCR, SDS-PAGE analysis and lytic activity of transformants of tobacco. The transformant having the highest lytic activity expressed about 40 micrograms of lysozyme per g of leaf tissue. PMID- 9739553 TI - Engineering of trypsin and its impact on beta-casein processing. AB - Tryptic processing of beta-casein yields several important nutraceutic and nutritious peptides. However, a final product peptide (1-105) stops the processing, inhibiting the enzyme. In attempt to modulate catalytic properties of this protease, K188 was replaced with aromatic amino acid residues. This aimed amplification of local hydrophobic and electrostatic interactions at the substrate binding site. The catalytic properties of obtained mutants (K188F, K188Y, and K188W) were measured at pH 7, 8, 9, and 10 with synthetic substrates and beta-casein. Kinetic analysis revealed that all the mutants conserve the capacity to split peptide bonds involving arginyl and lysyl residues. However, depending on mutation, the optimum pH of activity changes. As shown only by proteolysis of a natural substrate, produced mutants cleaved near 30 new peptide bonds compared to wild-type trypsin, 8 of them involving asparagine and glutamine amino acids. Some of the new cleavage sites can be related to the nature of the amino acid residue introduced in position 188. Consequently, only the joint use of several methods (synthetic substrate, protein substrate, influence of pH) can help to define better the differences of catalytic properties of wild-type and mutant proteases. Modifications introduced by the mutations are at the origin of the alteration of the specificity of the studied enzymes which are cleaving beta casein in many places, hydrolysing well the fragment 1-105. Since many tryptic inhibitors contain amidated Glu and Asp, and form amyloid structures, the new mutants could be used in hydrolysing resistant proteic structures. PMID- 9739554 TI - Building of a cation switch in trypsin (short communication). PMID- 9739555 TI - Construction of poly-met DNA yeast hybrids for increased methionine content: technofunctional properties of the hybrid yeasts. AB - Methionine is a limiting essential amino acid in human nutrition, to overcome the possible overdosage and improve bioavailability methionine supply should be in protein bound form. So insertion a poly-met encoding DNA sequence results a more efficient solution in increasing methionine content of yeast. Poly-met DNA yeast hybrids were constructed of Saccharomyces cerevisiae CB89, an auxotrophic mutant strain. Synthetic DNA sequence encoding methionine polypeptide was inserted into the polylinker region of pVT-U 100 vector with 2 mu plasmid replicon. After transformation of E. coli HB101 cells the efficiency of the ligation and transformation was checked by digesting the minipreps. S. cerevisiae CB89 was transformed with vector-poly-met insert and with the plasmid vector only as well. Hybrid yeasts were selected on uracilless medium. PVT-U 100 can be used as vector for the expression of DNA sequence in S. cerevisiae. The vector harbours the promoter of ADC1 gene immediately downstream from the promoter lies a polylinker sequence comprising unique restriction enzyme sites for BamHI, HindIII, PvuII, SacI, Xhol. The polylinker sequence is followed by the transcriptional stop site and polyadenylation signal of ADC1 gene. Plasmid pVT-U 100 has selection markers for S. cerevisiae (URA3) and for E. coli (amp, per F). Results show that the poly met DNA hybrids methionine content is influenced by the length of the insert. Fusion hybrids containing 600 bp oligo insert showed the best values. Distribution of methionine content in the protein subfractions of polymet DNA hybrid and parent strain CB89 was determined in dependence of glucose concentration and aeration intensity. The increase in synthesized methionine appeared in fractions 1 + 2 and residue. Technofunctional properties of parent strains and hybrids were compared for whole cells and cell wall (residue). Results demonstrate that enrichment in methionine in the cell wall fraction resulted improvement of emulsifying ability. Bioavailability of methionine content was better in DNA hybrid yeast than in parent strain and was the best when propagated in whey medium. PMID- 9739557 TI - Enzymatic phosphorylation of soy globulins by the protein kinase CK2. Determination of the phosphorylation sites of beta-conglycinin alpha subunit by mass spectrometry. AB - Beta-conglycinin alpha subunit has been phosphorylated using a cAMP-independent protein kinase (CK2) purified from the yeast Yarrowia lipolytica. CK2 is known to phosphorylate serines and threonines in the consensus sequence Ser/Thr-X-X Asp/Glu. Only 0.5 to 1 mol P/mol alpha subunit was incorporated although seven consensus sequences are present. Phosphorylated beta-conglycinin alpha subunit (P alpha) was digested by trypsin. The resulting peptides were analysed by RP-HPLC coupled to electrospray ionisation mass spectrometry (LC-ESMS). Two phosphopeptides were identified corresponding to 70-89 and 116-127 sequences with Ser 75 and Ser 117 phosphorylated respectively. Ser 75 is one of the predicted phosphorylation sites according to the consensus sequence criteria. Ser 117 is inside a very acidic peptide but does not belong to a previously described consensus sequence. PMID- 9739556 TI - Enzymatic phosphorylation of food proteins by purified and recombinant protein kinase CK2. AB - Protein kinase CK2 formerly called casein kinase II is a protein kinase able to phosphorylate more than 100 proteic substrates. We have purified protein kinase CK2 from the yeast Y. lipolytica to phosphorylate milk and plant reserve proteins to a significant extent. In the case of plant reserve proteins, which are polymeric substrates, not all subunits are substrate for protein kinase CK2, even if non phosphorylated subunits contain significant potent phosphorylations sites. Best substrates were soy beta-conglycinin (0.72 P/mol) and dephosphorylated caseins (0.5 P/mol). We have studied some functional properties of phosphorylated caseins. Solubility was improved for all pH values but pI. Sensitivity to calcium has also been assessed, and it is slightly improved upon phosphorylation. We have cloned the catalytic subunit of protein kinase CK2 from yeast Y. lipolytica. The recombinant catalytic subunit expressed in E. coli was active and displayed kinetic properties similar to those of the purified enzyme. The recombinant catalytic subunit was able to phosphorylate plant reserve proteins and milk proteins to a significant extent. Best substrates were soy beta-conglycinin (1.0 P/mol), and glycinin (0.59 P/mol). PMID- 9739558 TI - Crosslinking of sodium caseinate by a microbial transglutaminase. AB - Sodium caseinate is an effective substrate for transglutaminase. Crosslinking takes place at fast reaction rates resulting in high degrees of crosslinking. The polymers can be hydrolysed by trypsin, but the proteolysates contain residual portions of non- or partlyhydrolysed aggregates. Crosslinking of hydrolysed sodium caseinate decreased the number of free amino groups, but leads only to a very small increase in molecular weight of the peptides. PMID- 9739559 TI - Masking of antigen structure of soybean protein by conjugation with polysaccharide and cross-linkage with microbial transglutaminase (short communication). PMID- 9739560 TI - Substrate and binding specificity of aspartic proteases with milk clotting properties. AB - The hydrolysis of whole casein and isolated casein components were investigated with the purpose of obtaining information concerning the kinetic and specifty of aspartic proteases in rennin, pepsin and 4 microbial rennet substitutes. The velocity of hydrolysis decreased rapidly within the first hour. However, the hydrolysis was not completed after 2 days. A mathematical description of the slope of hydrolysis is possible by use of exponential equations. More than 40 peptides were detected by capillary electrophoresis or PAGE. The characterization of the C- and N-terminal amino acids of peptides shows that the hydrolysis of any peptide bond depends mainly on the structure of the C-terminal side chains of the amino acids. The detection of the basic amino acids lysin and arginin in the C terminal position of peptides is a new result, furthering the knowledge about the specificity of aspartic proteases. Differences in the reaction velocity or in the extent of hydrolysis are one of the possible explanations for the described differences in the rennet curd yield. It was concluded that the rennet enzymes are active also in the later phases of cheese ripening and are able to support the action of cheese ripening flora. PMID- 9739561 TI - Modifications of phosvitin by an immobilized protein phosphatase from Yarrowia lipolytica (short communication). PMID- 9739562 TI - Food proteins as precursors of peptides modulating human cell activity (short communication). PMID- 9739563 TI - Conformational transitions of holo-alpha-lactalbumin in hydro-ethanolic solutions. AB - Spectroscopic and thermodynamic studies of holo-alpha-lactalbumin folding show that in hydro-ethanolic media this protein structure is subjected to at least three distinct transitions induced by ethanol. As observed by spectrofluorescence and circular dichroism (CD) the first transition is only local, being associated with changes in the state of aromatic chromophores. During this transition overall tertiary structure of alpha-lactalbumin is retained. As shown by high sensitivity differential scanning calorimetry (HS-DSC) and CD, the second transition involves breakdown of the protein tertiary structure. The final organisation of the protein into highly helical folding may be considered as the third structural transition since the unfolding and the new helix formation are time-resolved processes. PMID- 9739564 TI - Effect of thermal denaturation on vanillin binding to some food proteins. AB - Interactions of food proteins--beta-lactoglobulin (BLG), bovine serum albumin (BSA) und ovalbumin (OA)--with vanillin and effect of thermal denaturation of the proteins on vanillin binding were studies by US-VIS spectrophotometry. This method has its origin in characteristic changes in the vanillin absorption spectrum at vanillin-protein complex formation and allows to calculate concentrations of the bound and free ligand in aqueous solutions. Thermodynamic parameters, the intrinsic association constants and the number of binding sites of the vanillin binding to the native and thermodenaturated proteins (monomers and clusters) were determined. It is shown that the vanillin affinity for the native proteins is decreased in the following order: BSA > BLG > OA. This sequence is reversed for the protein thermoclusters. The stepwise annealing allowing to derive complex protein mixtures composed of different types of the native and denatured protein mixtures composed of different types of the native and denatured protein particles was applied to thermodenaturation of BSA. The vanillin affinity for BSA is decreased in the order: native protein > denaturated monomer > denaturated clusters. Vanillin interaction with the proteins is mainly electrostatic in nature. PMID- 9739565 TI - On the novel catalytically-independent antimicrobial function of hen egg-white lysozyme: a conformation-dependent activity. AB - Dependency of the antimicrobial activity on the conformation of lysozme was examined by the means of gradual thermal inactivation at neutral pH and different temperatures. We found that heating of lysozyme at increasing temperatures for 20 min in pH 6.0 results in progressive loss of enzyme activity, while greatly promotes its antimicrobial action to the Gram-negative bacteria without a detrimental effect on the inherent bactericidal activity to Gram-positive ones, suggesting action independent of catalytic function. The most potent bactericidal conformation of lysozyme to either Gram-negative or -positie bacteria was that retaining approximately 50% of the native enzyme activity (HL80/6). HL80/6 showed several fold increase in surface hydrophobicity, with exposed two thiol groups, and 17% deamidation. Spectrophotometric analysis of HL80/6 revealed slight changes in its secondary structures, but considerable global conformational changes as a result of the formation of beta conformation, via cyclic imide, at the three aspartylglycyl sequences of lysozyme molecule. Direct damage to the bacertial membranes by HL80/6, was demonstrated by using ELISA and liposomal membrane model. Furthermore, the antimicrobial activity of HL80/6 was inhibited by the divalent cations Ca2+ and Mg2+ suggesting that HL80/6 interacts at a divalent cation binding site on the bacterial membrane and subsequently permeabilize it. The results introduce an interesting structure-antimicrobial relationship that the antimicrobial action of lysozyme is independent of its catalytic function. In addition, it is worth emphasizing that the naturally occurring conformational transition of lysozyme at physiological temperatures can be a biologically relevant event to switch its antimicrobial specificity to include the food-borne pathogens and heralding fascinating opportunities for application in formulated food systems. PMID- 9739566 TI - Physico-chemical modification of food proteins: food emulsions. AB - Physico-chemical protein modification involved in food processing is used for "engineering" the structure-property relationship of foods. The three main tools used for protein modification are food formulation, mechanical and thermal (heating/cooling) treatments. They correspond to the main stages of all food technologies. Food structural design is based upon incompatibility and complexing of food biopolymers as well as upon fundamental features of physico-chemical properties of biphasic aqueous systems. Some examples of functional biopolymer modification are considered in systems such as meat extenders, fat replacers, ice cream mixes and wheat flour doughs. PMID- 9739567 TI - Some physico-chemical properties and structural changes of bovine beta-casein upon glycation. AB - The studies on casein structure modification contribute to better understanding of the role of nonamino acid components in forming casein complexes and improving ways of protein functionality. The objective of the experiments was to explain the influence of bovine milk casein glycation on some physico-chemical properties and structural changes. From the the analysis of glycation rate curve the reaction of the first order range can be assumed during the first 24 h, turning to a mixed type afterwards. The isoelectric point and molecular weight of beta casein increased after glycation and the electrophoretic mobility was slightly modified. The structural changes were also confirmed by different absorption spectra in UV and a better heat stability of the modified beta-casein. The findings showed higher solubility with modified beta-casein. The glycation caused changes in beta-casein, modifying its susceptibility to the trypsin hydrolysis. PMID- 9739568 TI - Interaction of human serum albumin with dipalmitoylphosphatidylcholine in spread monolayers at the air/water interface (short communication). PMID- 9739570 TI - The effect of degree of whey protein denaturation and conditions of milk preparation on functional properties of yoghurt (short communication). PMID- 9739569 TI - Characteristics of heat-induced transparent gels from egg white by the addition of dextran sulfate and the protein-polysaccharide interactions (short communication). PMID- 9739572 TI - Schizophrenia: conceptual change in modern psychiatry. AB - Schizophrenia is perhaps the most devastating mental illness that psychiatrists must treat. Its chronic course with deteriorating consequences has been the focus of study throughout this century. This paper reviews the concept of schizophrenia, from fundamental Kraepelinian theory of 'dementia praecox' to biological correlates and explanation. Its causes, however, remain obscure and there were no appreciable changes in its treatment modality. A 'bio-psycho social' model of care is emphasized, aiming at the prevention of deterioration of symptoms, functional impairment, disability and social handicap. PMID- 9739571 TI - The structure-function relationship of ovalbumin matrix as the result of protein denaturation (short communication). PMID- 9739573 TI - Emil Kraepelin and modern psychiatry. AB - A brief account is given of the career and scientific contributions of the German psychiatrist, Emil Kraepelin, to whom the concept of schizophrenia is generally attributed. Attention is also drawn to his social and political outlook and its relationship to his work. The implications for contemporary psychiatry are outlined. PMID- 9739574 TI - Genetic epidemiology of schizophrenia: review and reassessment. AB - When a rigorous methodological approach is utilized, a substantial majority of recent studies provide evidence for the familial transmission of schizophrenia. Although the absolute rates of schizophrenia among relatives of schizophrenics tend to be lower than those reported in the earlier studies due to the restrictiveness of contemporary definitions of schizophrenia, the risk to relatives compared to that of controls has remained quite consistent. This observation that relatives of schizophrenics have an elevated risk for schizophrenia compared to controls is consistent with theories of both genetic and environmental transmission. Twin studies of schizophrenia have consistently reported greater concordance rates for monozygotic than dizygotic twins. Although this indicates the importance of genetic factors, the less than 100% concordance for monozygotic twins observed in every study indicates that nongenetic factors also play a role in the etiology of schizophrenia. Further, adoption studies offer an opportunity to unconfound genes and environment. The findings of adoption studies confirm that there are genetic components for schizophrenia. Even though we have shown that family, twin, and adoption studies have provided strong evidence for the role of genetic factors in schizophrenia, the mode of transmission remains unclear. The results of mathematical modeling studies do not support the single gene model. There is somewhat more support for the multifactorial polygenic model, but the model has also been rejected in several studies. Thus, the pattern of inheritance of schizophrenia has eluded an unambiguous characterization. Genetic linkage analysis promised to clarify the mechanisms of transmission, but early positive reports were subsequently overturned and, to date, there are no consistently replicated positive linkage findings for schizophrenia. There is now a world-wide search for the location of the genes on specific chromosomes which are responsible for schizophrenia. The clinical implications of current work to the future of locating a schizophrenic gene or genes will be discussed. PMID- 9739575 TI - Onset and course of the first schizophrenic episode. AB - Schizophrenia seems to occur with equal frequency in all countries and cultures. The ecological and social disparity of prevalence rates in open societies is not due to social causes, but to selection processes. At what point in the course of the disease social disadvantage begins and on account of which mechanisms is a question of both theoretical and practical importance. In a representative sample of 232 first episodes the true onset of schizophrenia is assessed by means of a standardized interview (IRAOS). The onset is 3 to 4 years later in women than in men, which is a result of the protective effect of estrogens as confirmed by animal experiments and by a controlled clinical study. For women the distribution of disease onset across the entire age range shows a delayed increase in adolescence and a second peak in the age group 40-45, presumably due to the fading effect of estrogen around menopause. For three quarters of all schizophrenias the onset lies between 15 and 30 years of age, the period of steepest social development. The distribution of the social biography through the psychosis, which occurs on average 3-4 years earlier in males than in females, determines the social starting conditions of the disorder and affects the early social course. Nearly 75% of the total cases begin with a prodromal phase of 5 years on average. The psychotic prephase lasts 1.1 years. The mainly negative prodromal symptoms are often associated with social and cognitive deficits already at this stage. In a case control study social stagnation or decline occur already during the prodromal phase. After the end of the first episode the mean values of social disability and the objective social status remain fairly stable over a period of at least three years. So far treatment and rehabilitation measures are not started until after first admission, which means after several years' duration of the disease when the majority of social deficits have already become manifest. PMID- 9739576 TI - The social course of schizophrenia: local and societal factors. AB - In this paper, we propose a model of social course of schizophrenia based on cross-cultural research on the influence of family, wider social network, work, political economy, and legal and mental health care institutions on the experience of illness. We posit the way these ordinary arrangements of daily living organize the course of schizophrenia in part through cultural processes that affect the body-self in suffering and in part through social processes that establish an intersubjective matrix for the experience of illness. We believe this model can be generalized to other chronic illness such as depression, diabetes, asthma, osteoarthritis, chronic pain syndrome, chronic fatigue syndrome, and even heart disease and cancer. We develop the implications of this anthropological approach for research and practice. PMID- 9739577 TI - Pharmacological prevention of relapse. AB - Relapse is the "return of a disease after partial recovery", and is a major feature of schizophrenia disorder. It can be defined in terms of need for change in treatment, including rehospitalization or crisis intervention, the re emergence of florid psychotic features, or gross social decompensation. Relapse is best viewed as continuum of severity rather than as discrete "attacks". Factors influencing relapse include major life events and the family constellation. Antipsychotic drugs protect against the latter but not the former, and relapse may be mediated by non-specific arousal mechanisms. The efficacy of drug treatment in postponing rather than preventing relapse is well established. The interval between relapses is prolonged at least two-fold, but in the long run most patients relapse. Unwanted effects of antipsychotic drugs can be a burden to patients, impairing quality of life. In particular, movement disorders and subjective dysphoria may be marked, as may compliance. Of these EPS, tardive dyskinesia is the most serious on long term use. Non-EPS long term effects include weight gain and endocrine changes. Depot medication has advantages over oral medication in the more ill, less compliant patients. Side effects may, however, be more marked. The greatest pain is in improved compliance but the regular supervision of the patient is also helpful. Pharmacokinetic issues are poorly understood. High and mega-dose strategies have been advocated. High doses may be needed in some patients, but megadoses are rarely justified and may be hazardous. Low dose and intermittent therapy have been evaluated but are not as successful as hoped. Some less ill patients may benefit. These schedules depend on the identification of prodromata of relapse which is not always easy, nor are relapses necessarily preceded by prodromata. Newer drugs are being developed rapidly in the search for a safer clozapine, the only antipsychotic with definitely enhanced efficacy. Other drugs which have been re-evaluated include the benzodiazepines. However, the area of greatest priority in research is that of interactions, particularly potentiation, between drug and non-drug treatments. PMID- 9739578 TI - [Chemotherapy for tuberculosis; yesterday, today, and tomorrow--basic and clinical studies]. AB - The discovery of streptomycin in 1944 had given rise to great flowering of chemotherapy for tuberculosis. The times which triple treatment of SM.PAS.INH after the temporal time of SM.PAS had been standard regimens on initial treatment had continued for more than twenty years. The shortening of duration for chemotherapy had become possible by the introduction of RFP, and the duration had reduced to one fourth compared with that of the regimens till then by the addition of PZA for two months at the beginning of treatment on the initial treatment cases. In this paper, historical aspects of early and present-day chemotherapy of tuberculosis and the reports of main studies have been summarized, and pharmacokinetics of INH, action of antituberculous drugs in short course chemotherapy, MDR-TB and biological response modifiers for treatment of tuberculosis, etc. has been reviewed. It is urgently awaited that more new drugs without cross resistance to previous drugs will be developed for the more shortening of the duration and the improvement of the treatment for MDR-TB. PMID- 9739580 TI - [Time trend in incidence and mortality of tuberculosis and characteristics of notified tuberculosis patients in urban area of Mongolia]. AB - Recent Mongolian political, social and economic changes have had a great impact on its health care system and tuberculosis control program. The objective of this study is to assess time trend in incidence and mortality of tuberculosis and characteristics of notified tuberculosis cases in Mongolia. 1) Data on statistics of tuberculosis are obtained from reports of the National Tuberculosis Center in Mongolia. The mortality of tuberculosis in Mongolia shows a downward trend during 1985-1995. The number of notified tuberculosis cases had gradually decreased during 1985-1989. It suddenly dropped in 1990 and was the lowest in 1993. After that, about two fold increase in the notified cases was observed in recent three years from 1993 to 1995. Such a large fluctuation in the number of notified cases after 1990 is unlikely to be associated with the epidemiologic situation of tuberculosis, but rather due to a reporting bias. The shortage of drugs and economic hardship prevented patients from consulting medical facilities. The shortage of drugs also prevented doctors from notifying patients to the tuberculosis registry, because the notification did not lead to treating the disease. The improvement of health care system and the supply of essential drugs since 1994 seems to contribute to the increase in the number of notified cases. 2) The study subjects include 618 patients who were diagnosed as active tuberculosis at ten tuberculosis specialized facilities in Ulaanbaatar, Mongolia from May 1995 to March 1996. Patients were interviewed about their demographic factors and their medical records were reviewed. Fifty one percent of the cases were female. The mean age was 26.9 years old. Ninety percent of the cases underwent chest X-ray examination, while 72% of the cases underwent bacteriological examination and only 21% were confirmed bacteriologically. It is necessary to improve the quality control of sputum smear examination and the validity of diagnosis of tuberculosis in Mongolia. As for treatment regimens, only 29% of the cases were being treated with at least four drugs (isoniazid, rifampicin, pyrazinamide, ethambutol and/or streptomycin). It is needed to provide directly observed treatment using the WHO recommended standard regimen to at least smear positive tuberculosis cases. PMID- 9739579 TI - [Attributable factors to the emergence of multidrug-resistant Mycobacterium tuberculosis based on the observation of consecutive drug resistance test results]. AB - Thirty six cases with multidrug-resistant tuberculosis were retrospectively studied to define the causes attributable to the emergence of multidrug-resistant M. tuberculosis. All these tuberculosis cases were microbiologically confirmed and resistant to at least isoniazid and rifampicin. Data analysis using matched pair sampling methods (1:3) demonstrated that the followings are the significant risk factors for the emergence of multidrug-resistant tuberculosis; incompliance to treatment (Odds ratio 21.0: 95% CI 4.10-107.63), alcohol abuse (Odds ratio 15.0: 95% CI 2.34-96.1) and the history of previous treatment (Odds ratio 5.0: 95% CI 2.04-12.21), while diabetes mellitus is not statistically significant. The incompliance to treatment which is primarily thought to be patient's responsibility results in non-optimal administration of antituberculous agents, leading to the multidrug-resistant tuberculosis. Other factors that may have contributed to the emergence of resistance included the unnecessary change of regimen before completion of chemotherapy. This is patient-unrelated situation where responsibility lies in the medical side. A clinical case presented here is an example. In this case RFP was replaced with ethambutol 3-months after the initiation of regimen including SM, INH and RFP because of abnormal elevation of GOT and GPT without any supporting evidence that RFP was causative. The readministration of RFP after 1-year cessation did not induce liver dysfunction, while the drug resistance was observed not only to RFP but also to INH. This case suggests unnecessary interruption of RFP could lead to the emergence of resistance to INH as well as RFP. One known mechanism of drug resistance is random mutation and the selection by drugs administered during the course of chemotherapy. The cases with advanced cavitary lesions would have a higher probability of the occurrence of mutation. The more the number of mutant bacilli, the higher the probability of emergence of multidrug resistance. Those cases in which longer period of time is needed for the negative conversion of M. tuberculosis should be treated with potent chemotherapy regimens under the intense supervision. Since both INH and RFP are the most potent among currently available antituberculous agents. It is crucial to preserve the potency of these essential agents before novel antituberculous are developed. PMID- 9739581 TI - [Adverse reactions of antituberculous agents]. AB - We experienced adverse reactions to antituberculous agents in 17 patients (53%) out of 32 patients treated for tuberculosis and nontuberculous pulmonary mycobacteriosis. Side effects were seen in 15 patients (47%), and abnormal laboratory findings were observed in 9 patients (28%). Most side effects mostly appeared within two weeks after the administration, and were of short duration. However, there were also side effects such as neurological symptoms, arthralgia, and general fatigue which appeared after one month and lasted for a long duration. These results suggest that careful observation for adverse reactions in antituberculous treatment is required. Finally, 11 patients with adverse reactions were successfully treated by changing antituberculous agents or readministration after temporary stop of the administration. Four patients improved even with continued use of agents causing side-effects. However, in the case of two patients who had to change antituberculous agents, adverse reactions to all antituberculous agents appeared and the treatment for tuberculosis and nontuberculous pulmonary mycobacteriosis had to be stopped. PMID- 9739582 TI - [A case of AIDS with pleuropulmonary tuberculosis in which PCR was useful in making definitive diagnosis]. PMID- 9739583 TI - [Bone mineral density (BMD) of the calcaneus bone in 72 dialysis patients measured by ultrasonic bone absorptiometry]. AB - BACKGROUND: To determine what factors contribute to and change BMD in dialysis patients. METHODS: Bone parameters, namely, osteosono-assessment index (calculated by speed of sound and transmission index) were measured at calcaneus bone by ultrasonic bone absorptiometry with an Aloka Acoustic Osteoscreener (Model AOS-100). Seventy two patients were 62. 8 +/- 10.5 years of age (mean +/- SD) (range 36-81) and 70.8% were males; the patients had received dialytic therapy for 52.7 +/- 41.7 months (range 2-205). The effects of sex, age, height, weight, postmenoposal years, dialysis duration, various blood parameters, oral phosphorus binding agent (CaCO3 dosage) and oral vitamin D3 treatment (D3 dosage) on BMD were assessed statistically. RESULT: BMD in male patients were significantly higher than those in female patients (Mann-Whitney test, p < 0.01). BMD showed significant correlations with age (Spearman's correlation coefficient r = -0.384, p < 0.01), height (r = 0.479, p < 0.00001), postmenoposal years (r = 0.692, p < 0.05), dialysis duration (r = 0.250, p < 0.01), blood Ca level (r = 0.292, p < 0.05) and CaCO3 dosage, postmenoposal years and dialysis duration were the variables that significantly correlated with the BMD (multiple correlation coefficient = 0.646). PMID- 9739584 TI - [Operative methods for severe hypospadias]. AB - BACKGROUND: Pediatric urologists tend to use one-stage procedures for the repair of hypospadias. As there are various types of hypospadias, we cannot repair this disease with a single modality. It is difficult to estimate the exact length of the neourethra in cases of severe hypospadias before surgery. METHODS: After a circumferential incision is made about the coronal sulcus and the chordee is completely released, the distance between the glans tip and the retreated native meatus is measured to determine the length of the neourethra. Urethroplasty with the method of Transverse Preputial Island Flap (TPIF) is selected when the distance ranges from 3 to 4 cm, while urethroplasty using with modified OUPF IV (Koyanagi) is selected in cases of more than 4 cm. RESULTS: We performed surgery on 14 hypospadiac patients with chordee between April 1996 and April 1997. Eight patients underwent urethroplasty using the TPIF method and 6 underwent urethroplasty with the method of the modified OUPF IV. With the TPIF methods, 7 to 8 patients underwent repair successfully and one experienced urethrocutaneous fistula, while 5 of 6 treated by the modified OUPF IV method has successful repairs and meatal stenosis occurred in one patient. CONCLUSION: Even if we encounter severe hypospadias, we can treat these patients with one stage repair alternatively. A relatively high success rate was obtained with both methods to repair severe hypospadias. PMID- 9739585 TI - [Direct effects of Chinese herbal medicine "hochuekkito" on sperm movement]. AB - BACKGROUND AND PURPOSE: Chinese herbal medicine, "Hochuekkito" is widely used for male infertility in Japan. There have been many reports concerning its clinical usefulness but very few reports of in vitro experiments studying the mechanism of its effects. In addition to stimulating germ cells, we analyzed its direct effects on sperm using computer assisted semen analyzer (CASA). MATERIALS AND METHODS: Motile sperm were prepared using swim up technique from semen collected from ten healthy volunteers. Sperm movements (motility, velocity, linearity) were analyzed by CASA after adding either serum containing anti-sperm antibody (ASA) or normal serum with or without Hochuekkito. RESULTS: Two hours after adding serum with ASA, the decrease of sperm motility was significantly reduced from 25.1% (92.8%-->67.7%) to 12.5% (92.9%-->80.6%) by adding Hochuekkito. No significant difference in velocity and linearity was observed between two groups. By adding normal serum, any of three parameters differed significantly with or without Hochuekkito. CONCLUSION: Protective effects of Hochuekkito on sperm was suggested. Although normal sperm with ASA was used in this report, since the sperm of infertile patients are said to be more fragile, this results imply that direct protective effect is one of the mechanism of Hochuekkito for male infertility. PMID- 9739586 TI - [Prognosis in the cases with renal cell carcinoma according to clinical parameters]. AB - BACKGROUND: The objects of this study is to evaluate the clinical prognostic factors in renal cell carcinoma. MATERIALS AND METHODS: During a 30-year period from January 1965 to December 1994, 1301 cases with renal cell carcinoma were treated at the Yokohama City University Hospital and its affiliated hospitals. In these cases, cause specific 679 cases from January 1965 to December 1990 were analyzed in a study undertaken to investigate long-term treatment results and clinical prognostic factors. RESULTS: 1. The cause specific 5-, 10-, 15-, and 20 year survival rates were 48.7%, 41.1%, 32.3%, and 26.5% respectively, indicating thus that a great number of cases had an ominous prognosis even 5 years or moreafter surgical treatment. 2. Among patients under 40 years of age (n = 29) none died more than 2 years after receiving operation, the prognosis for this particular group of cases being relatively good. 3. Female, incidentally detected cancer, small tumor size (< or = 4.0 cm), slow growing type and low stage were proven to be favourable prognostic factors in renal cell carcinoma. 4. The cause specific 5-year survival rate for the patients (n = 239) from 1965 to 1981 was 33.8%, while the rate for the patients (n = 440) from 1982 to 1990 was 56.5%. This improvement of survival rate was brought by the increase of the incidentally detected renal cell carcinoma. 5. In the incidentally detected renal cell carcinoma, the incidence of slow growing cases and the cases of less than 4.0 cm tumor size were higher than in the symptomatic renal cell carcinoma. 6. Multivariate analysis using Cox's proportional hazard model showed that stage was the most important prognostic factor. CONCLUSIONS: These results suggested that sex, age, symptom, tumor size, growing type, and stage were important prognostic factors in renal cell carcinoma. PMID- 9739588 TI - [Acupuncture for urinary incontinence in patients with chronic spinal cord injury. A preliminary report]. AB - PURPOSE: We investigated the possible use of acupuncture for the treatment of urinary incontinence caused by detrusor hyperflexia in patients with chronic spinal cord injury. METHOD: A total of 8 male chronic spinal cord injured patients with urinary incontinence were treated by acupuncture. Their ages ranged from 20 to 33 years (mean 27). The level of lesion was cervical in 4 and thoracic in 4. Detrusor hyperreflexia with uninhibited bladder contraction was confirmed by urodynamic studies in all of them. Acupuncture was performed using a disposable stainless needle (0.3 mm in diameter, 60 mm in length), which was inserted into bilateral BL-33 (Zhongliao) points and was rotated manually for 10 minutes. The treatment was conducted every week for 4 weeks. Urodynamic studies were repeated, immediately after the beginning of and a week after the completion of the treatment. Urinary symptoms were also checked before and after the treatment. RESULTS: No side effects were recognized throughout the treatment period. Among 8 patients, incontinence was controlled completely in 3 (38%) and partially in 3 (38%). The average maximum cystometric bladder capacity increased significantly, from 42.3 +/- 37.9 ml to 148.1 +/- 101.2 ml by the treatment (p < 0.05), while the average maximum bladder pressure was not changed. CONCLUSIONS: These data suggest that acupuncture could be a promising alternative for conventional therapies for urinary incontinence caused by detrusor hyperreflexia in patients with chronic spinal cord injuries. PMID- 9739587 TI - [Combination chemotherapy with cis-platinum and ifosfamide for hormone unresponsive prostate cancer]. AB - PURPOSE: There is no effective therapy against hormone refractory prostate cancer. This led us to evaluate the effectiveness and toxicity of cis-platinum (CDDP) and ifosfamide (IFM) combination chemotherapy in the patients with hormone unresponsive carcinoma of the prostate. METHODS: Patients with hormone unresponsive prostate cancer were scheduled to receive CDDP 70 mg/m2 intravenously on day 1 and IFM 1.2 g/m2/day intravenously on day 1 through day 5 of 28-day cycle. RESULTS: Twenty seven patients with hormone unresponsive prostate cancer were enrolled onto this trial. Of these patients, seven (26%) demonstrated a partial objective response (PR), and ten (37%) a stable disease (ST). The response duration of PR cases lasted from 6 to 49 months with a median of 16 months and the response duration of PR + ST cases lasted from 3 to 36 months with a median of 10 months. Subjective improvement was obtained in 11 patients (41%). Survival duration of all cases were 4 to 89 months with a median of 23 months and probabilities of survival at 3 years and 5 years were 36% and 24%, respectively. The toxicity of this treatment was mostly mild to moderate, anemia (96%), leukocytopenia (89%), anorexia (81%), alopecia (67%), thrombocytopenia (44%), hematuria (38%), renal dysfunction (19%) and liver dysfunction (7%) were noticed. Severe toxicity was observed in two cases, one acute renal failure and one endotoxin shock. CONCLUSION: We conclude that CDDP and IFM combination chemotherapy was active regimen for hormone unresponsive prostate cancer. PMID- 9739589 TI - [Adenoid cystic carcinoma of the prostate. A case report]. AB - Adenoid cystic carcinoma is most commonly encountered in the salivary glands. It is rarely found in the prostate but distinctive variant of prostatic adenocarcinoma. This case report is thought to be the first one in the Japanese literature. A 51-year-old man was admitted with urinary difficulties and frequency since one month. Digital rectal examination revealed an enlarged stony hard prostate. The transurethral prostatectomy was performed and the histopathological diagnosis was the adenoid cystic carcinoma. PSA and PAP were normal. The findings of computerized tomography suggested the invasion to the bladder and rectum. Therefore the total pelvic excenteration was carried out. He subsequently received radiation therapy (50 Gy to the pelvis), anti-androgen therapy and three courses of chemotherapy using Cisplatin, Peplomycin, Epirubicin and so on. But the metastatic disease involving the liver and pelvic lymph nodes developed, and he died two years seven months postoperatively. PMID- 9739590 TI - [Cyclophosphamide induced urinary bladder and renal pelvic tumor--a case report]. AB - We present a case of asynchronous development of transitional cell carcinoma in urinary bladder and renal pelvis after prolonged cyclophosphamide therapy. A 57 year-old woman had received 290 g cyclophosphamide for 13 years because of therapy for non-Hodgkin lymphoma. She was suffered from dysuria and macrohematuria and visited our clinic. Cystoscopy, CT and MRI revealed invasive bladder tumor and total cystectomy was performed. Histological diagnosis was transitional cell carcinoma, G3 < G2, pT4. Six months after the cystectomy, a follow up urography and computerized tomography showed left renal pelvic tumor. The patient underwent total nephroureterectomy, and the histological diagnosis was transitional cell carcinoma, G3, pT3. We reviewed cyclophosphamide induced urothelial carcinomas from Japanese and world literatures. PMID- 9739591 TI - [Normal term delivery after adrenalectomy for Cushing syndrome in a pregnant patient--a case report]. AB - This is a case of a 29-year-old female. She visited our hospital with the chief complaint of swelling of the legs and abdominal bloating at the 10th week of pregnancy. Edema and central obesity were observed in the lower extremities. Hematological and biochemical tests revealed hypokalemia. Gynecologically, she was normal. Elevated blood cortisol levels were identified on the hormonal tests, which suggested the existence of Cushing's syndrome. Ultrasound revealed the presence of a tumor in the left adrenal gland, and she was referred to our department for surgery. On September 13, 1994, left adrenalectomy was performed in a right lateral position under inhalation anesthesia and epidural anesthesia. Adjuvand steroid therapy was initiated during surgery and the blood cortisol levels were normalized within 2 months. She delivered a baby girl weighing 2,722 g at the 40th week of pregnancy via a normal transvaginal delivery. Pregnancy rarely occurs in patients with Cushing's syndrome. We think she was the first case in Japan who had normal 40th week transvaginal delivery after adrenalectomy during pregnancy. We present a description of our case here with reference to the relevant literature. PMID- 9739592 TI - [Blood products use and related factors in Japan]. AB - PURPOSES: To examine the use of blood products in Japan by prefectures and to determine the factors associated with the amount of blood products used. METHODS: The amounts of blood products used in November 1995 were surveyed in 8,723 hospitals throughout Japan. The survey also included whether the blood products in hospitals were stored and managed based on the guidelines for appropriate use of blood products. RESULTS: 1. 4,675(90.9%) of the 5,141 respondents used blood products in 1995. 2. The amount of use of each blood product was considerably different among the prefectures. Larger amounts of red blood products and plasma products were used in urban areas. 3. The amounts of use of red blood cell products, plasma products and platelet concentrates showed similar geographical distributions at a significant correlation (r > or = 0.6) each other. The amount of use of albumin products correlated significantly (r = 0.69) with that of immunoglobulin products. 4. Among the various blood products surveyed in this study, the amount of use of only red blood cell products was comparatively larger in the prefectures where blood products were appropriately stored and managed. 5. The number of surgical operations performed under general anesthetization correlated positively with the amount of use of red blood cell products, plasma products and platelet concentrates; a similar correlation was observed between the number of operations for malignant neoplasm and cardiac diseases. CONCLUSION: The amounts of use of blood products used were significantly different among the prefectures in Japan. The amount of use of red blood cell products possibly reflects demand and consumption of blood products as a whole. PMID- 9739593 TI - [Functional fitness and related factors in community-dwelling elderly]. AB - To examine the association of level of functional fitness to demographic, health, and life behavioral or social factors, cross sectional data were obtained for 737 persons aged 60 years or older, and who were independently living in the community. Functional fitness was measured with a functional fitness test containing 4 task items: standing, walking, hand performance, and self-care performance. Among the demographic factors, statistically significant associations with functional fitness were found for age in both male and female and for the presence of spouse in male. Health status, previous or present history of circulatory diseases and musculo-skeletal diseases were significantly associated with lower levels of functional fitness in male, and previous or present history of musculo-skeletal diseases and presence of higher obesity associated with lower fitness level in female. With life behaviors, men who had habitual exercise activities and women who had no habitual nap but habitual exercise activities and frequent out-of-home activities showed significantly higher fitness level than their counterparts. These results suggest that level of functional fitness in independently living aged people in the community was significantly associated with the presence of spouse, history of circulatory and musculo-skeletal diseases, and habitual exercise activities in males; and with the history of musculo-skeletal diseases, obesity, and habitual exercise activities, napping, and frequent out-of-home activities in females. PMID- 9739594 TI - [Factors aggravating bronchial asthma in urban children (I)--The involvement of indoor air pollution]. AB - The aggravation of bronchial asthma in today's urban child population was studied by an epidemiological study in order to elucidate the involvement of indoor air pollution in relation to housing style. The asthma group consisted of 210 children under 12 years old who had been recently diagnosed as having bronchial asthma and under the care of Osaka Prefectural Habikino Hospital. The non-asthma group consisted of 180 children under 12 years old who had been under care at Osaka Prefectural Hospital but had no present history of allergic symptom. The individual atmospheric environment and housing style were surveyed by questionnaire. Also, the amount of mite allergen (Dp: Dermatophagoides pteronyssinus, Df: Dermatophagoides farinae) in room and bedding dust and the concentration of cotinine in urine were examined as objective indicators for the load of environmental allergen and the indoor air pollution by tobacco smoke, respectively. In this study, bronchial asthma was classified into two types: atopic/non-atopic, according to whether Dp-specific immunoglobulin E (Dp-IgE) was present/absent (positive/negative). Thus, for the risk factors given above, their involvement in each type of asthma was examined by comparing the proportion of the exposed subjects between the three groups of atopic asthma, non-atopic asthma and non-asthma. As atopy is an important factor of child asthma, the relative risk (odds ratio) of Dp-IgE increase (atopy) was also determined for the same factors by logistic regression analysis in each of the asthma and non-asthma groups. The results are as follows: 1. Reinforced concrete housing material, which results in mal-ventilation, increased the load of indoor air pollutants such as tobacco smoke. 2. A higher amount of mite allergen in bedding dust increased Dp-IgE. 3. Heating with stove, which results in a higher room humidity as well as temperature, enhanced Dp proliferation and appeared to be involved in increasing the risk of atopic asthma. 4. Reinforced concrete housing material appeared to be involved in suppressing Dp-IgE and increasing the risk of non atopic asthma. But some air pollutants such as tobacco smoke and mite allergen showed no relationship to these involvements. PMID- 9739595 TI - [Factors aggravating bronchial asthma in urban children (II)--The involvement of atopy and serum fatty acids, and their interaction with urban living environment]. AB - The aggravation of bronchial asthma in today's urban child population was studied by an epidemiological study in order to elucidate the involvement of food habits as well as individual factors such as age, sex, and history of atopic dermatitis, and the interaction with urban living environments. The asthma group consisted of 202 children under 12 years old who had been recently diagnosed as having bronchial asthma and under the care of Osaka Prefectural Habikino Hospital. The non-asthma group consisted of 81 children under 12 years old who had been under care at Osaka Prefectural Hospital but had no present history of allergic symptom. The individual factors and the urban living environments (atmospheric environment, housing style) were surveyed by questionnaire. Also, the mite (Dp: Dermatophagoides pteronyssinus, Df: Dermatophagoides farinae) specific immunoglobulin E (Dp-IgE, Df-IgE) and the composition of serum fatty acid were examined as objective indicators for atopy and dietary habits, respectively. In this study, bronchial asthma was classified into two types: atopic/non-atopic, according to whether Dp-IgE was present/absent (positive/negative). Thus, for the risk factors given above, their involvement in each type of asthma was examined. As atopy is an important factor of child asthma, the relative risk (odds ratio) of Dp-IgE increase (atopy) was also examined by logistic regression analysis in each of the asthma and non-asthma groups. The results are as follows: 1. As age increased, the risk of atopy increased but the risk of asthma decreased. 2. The risk of asthma increased as Dp-IgE rather than Df-IgE increased. 3. For the composition of serum fatty acid, the lower quartile ranking (LQR) group for a saturated fatty acid, stearic acid, and the upper quartile ranking (UQR) group for a mono-unsaturated fatty acid, oleic acid, had a higher risk of nonatopic asthma. 4. The LQR group for omega 6-poly-unsaturated fatty acid such as linoleic acid, had a lower risk of atopy. 5. The UQR group for a omega 3-poly-unsaturated fatty acid, eicosapentaenoic acid, had a higher risk of nonatopic asthma. The UQR of eicosapentaenoic acid and living environments within 25 m from a major road or housing of reinforced concrete structure showed involvement in synergistic increase in the risk of non-atopic asthma. PMID- 9739596 TI - [Study on abuse and neglect of the disabled elderly living at home]. AB - To obtain basic information on elderly abuse and neglect in order to provide for its early discovery and prevention, a questionnaire survey about the present situation regarding abuse and neglect of the disabled elderly living at home was performed on health/medical service/welfare professions. Forty-two cases were analyzed. The major results were as follows; 1. Victims were 13 men and 29 women, approximately 1/3 of whom were in their 70's and another 1/3 in their 80's. Approximately 1/4 of the abusers were either sons or daughters of the victims. 2. Verbal abuse was the most frequent type of abuse with a rate of 69.0%. The percentage of psychological abuse was 61.9, passive neglect was 57.1%, active neglect was 50.0%, physical abuse was 47.6%, passive self-neglect was 28.6%, financial/material exploitation was 23.8%, active self-neglect was 16.7% and the others was 11.9%. The average number of abuse and neglect that an elderly received was 3.5. 3. The main causes of abuse and neglect did not appear to be simple but was complicated by related causes, many of both victims and abusers had elements in their personality, developmental history and interpersonal relationships that over a period of years formed the basis for the problematic behavior. Many abuse or neglect cases arose from caregiving burden of family caregivers or insufficient social systems for support to meet the needs of caring for the elderly. 4. The results suggest that for prevention and countermeasures for elderly abuse, there is an urgent need to arrange for expansion of health and welfare services to reduce burden of caregivers, provide for education of professionals of health/medical service/welfare for early discovery and proper handling of abuse problems, development of a checklist for early discovery, expansion of opportunities of improving care skill for family caregivers and establishment of consultation system for the elderly and caregivers, and organizing emergency shelter for victims of elderly abuse. PMID- 9739597 TI - [Discussing AIDS with their junior high school children by parents. A study on parents education about AIDS]. AB - PURPOSE: This study was performed to determine the extent of parental participation in talking to their children regarding AIDS, and to determine what intervening factors existed. FOCUS GROUP: Parents of children who attend a public junior high school in Yokohama, Japan. TERM: November 9, 1995 to November 16, 1995. METHOD: Anonymous questionnaires addressed to both parents were distributed to children in the classroom. After the parents had completed the form, the children brought it back to the school. PARTICIPANTS: 616 Families or 1,117 individuals (509 fathers, 608 mothers) Response Rate: 72.4%. RESULTS: 1. Percent of parents who have talked to their children about AIDS (Experience Rate): Fathers 26%, Mothers 63%. 2. Percent of parents willing to discuss AIDS with their children (Willingness Rate): Fathers 49%, Mothers 70%. 3. In 70% of the families either the father or mother have talked with their children about AIDS (Experience Rate). In 95% of the families either the father or mother expressed a willingness to talk with their children about AIDS (Willingness Rate). 4. Factors related to Experience Rate were: Fathers--No consistent pattern was shown in responses. Mothers--1. Have enough knowledge about AIDS. 2. Understand basic facts about the route of infection of HIV. 3. Have expectations of an increase in the number of PWA/H (People with AIDS/HIV) in the near future. 4. Recognize the risk of HIV infection for their children. 5. Factors related to Willingness Rate were: Fathers--1. Understand that AIDS is not only someone else's problem. 2. Recognize the risk of HIV infection for their children. Mothers--1. Understand that AIDS is not only someone else's problem. 2. Have an accepting attitude toward their children's friend who is infected with HIV. PMID- 9739598 TI - [Lifestyles in participants at health screening test]. PMID- 9739599 TI - [The current situation of HIV/AIDS epidemic in Thailand. The importance of the relationship between seroepidemiological survey and behavior survey]. PMID- 9739600 TI - [Registration of cause of death in Denmark]. PMID- 9739601 TI - [Cross-sectional therapeutic programs--an example of a cooperative health care system. A review with comments]. AB - There is no tradition for sharing the responsibility for episodes of care between the primary and secondary sectors in the Danish health care system. Concurrently with increased international experience with shared care programmes, there is also a growing interest in Denmark in cooperation between the sectors. Based on literature research, shared care programmes are presented as a method of ensuring continuity and quality in treatment of chronic diseases. Experiences in the areas of diabetes, asthma, rheumatoid arthritis, and cancer are described. It is concluded that the Danish health care system is well prepared for the implementation of shared care programmes; there are only few sources of payment in the system, and an extensive continuing medical education system ensures that general practitioners can participate in relevant education. The implementation of shared care programmes in Denmark should be followed by scientific evaluation and documentation of the quality of the treatment programmes. PMID- 9739602 TI - [DNA biobanks. Establishment and maintenance]. AB - The need for extraction, purification and storage of DNA in biobanks is increasing. DNA may be obtained from mouth brush water, guthrie cards, tissue biopsies or venous blood. Sampling conditions depend on the method used for procurement of DNA, e.g. DNA extraction or Ebstein Barr Virus transformation. Exact knowledge about the validity and stability of DNA stored in buffer is still insufficient. Biobanks at hospitals and at research departments are regulated by the Danish Private Registers, etc. Act. Research projects based on DNA biobanks should be notified to the Danish Data Protection Agency and approved by the local ethical committee. Discount, economic, and business class set-ups are different practical and financial models for the structure of DNA biobanking. PMID- 9739603 TI - [Reliability of death certificates. The reproducibility of the recorded causes of death in patients admitted to departments of internal medicine]. AB - The aim of the study was to assess the reproducibility when different doctors fill in diagnoses on death certificates. Records from 40 patients who had died in 1994 during admission to a medical hospital department in Denmark were selected at random. Ten doctors filled in a death certificate for each patient (without knowledge of autopsy findings and results of examinations received after the patient's death). The agreement between the diagnoses was assessed using a rating scale with seven categories. Afterwards the 400 death certificates were mixed and coded by the Medicostatistical Section of the Danish National Board of Health. The diagnoses made by the ten doctors showed insignificant discrepancies in ten cases, larger discrepancies were found in eight cases and large discrepancies in 19 cases. In three cases the patient had died suddenly and little information was available. The coding was standardized at the Danish National Board of Health, but their diagnoses reflected the discrepancies between the doctors' diagnoses. In conclusion, the reproducibility of diagnoses on death certificates is so poor that information from the Registry of Causes of Death is of little use of administrative or scientific purposes. PMID- 9739604 TI - [The illicit drug market in Aarhus. I. Supply]. AB - Drugs (n = 2240) seized on the illicit drug market in the Aarhus Police District during the period 1.1.1992-31.12.1993 are described with regard to weight, identity, purity and distribution. This article dealing with drugs together with the article dealing with the "actors" are reviews of results published in book form (English summary) (1). Two thirds of the samples were cannabis, mainly hash. Medical prescription drugs, mainly benzodiazepines, constituted 11% of the samples. Amphetamine was found in 9% of the samples and heroin in 8%. Cocaine and other euphoriants were each found in less than 1% of the samples. The purity of all types of drugs varied greatly during the study period, good and poor quality drugs being available on the market at the same time. No difference was found between the purity of drugs in user packages and that of drugs in large packages. Most batches were on the market only for a brief period of time. It is concluded that it is difficult for users to obtain the same quality of a substance each time. PMID- 9739605 TI - [The illicit drug market in Aarhus. II. Actors]. AB - People (n = 823) from whom drugs were confiscated by the police during the period 01.01.1992-31.12.1993 are characterized with regard to sex, age, nationality, other criminal offences etc. The data were obtained from the Central Criminal Register, the database of the National Drugs Intelligence Unit and the Danish Board of Health methadone list. Seven hundred and fifty men and 73 women were charged with drug offences in Aarhus during the two-year period. Fifty of these were foreign nationals. Almost all of the suspects were drug abusers. Ten percent of the people in the study were on methadone treatment at the time of confiscation. Sixty percent were categorized as users of cannabis only, 16% used heroin, 16% stimulants, and 7% medical drugs. Almost all of the suspects in the study were registered for offences other than drug offences (mainly theft and burglary). It is concluded that a substantial number of drug addicts in methadone treatment were found in possession of illicit drugs, and that most of the people charged with drug offences had previously been charged with other criminal offences. PMID- 9739606 TI - [Assessment of medical technology in practice]. AB - With the establishment of a national institute for Health Technology Assessment (HTA), the interest in HTA is increasing in Denmark. The National Board of Health defines HTA as a comprehensive systematic evaluation of the assumptions for, and consequences of, the application of health technology. The focus is on four elements: the technology, the economy, the patient and the organisation. However, is this broad and comprehensive definition in agreement with the practical use of HTA? This article refers to an international comparison of 124 HTA-projects made by five national HTA-institutions. The article shows that only seventeen HTA projects can be characterized as broad and comprehensive, focusing on all four elements. The rest are more restricted in their form. The future implication for Danish HTA initiatives might then be to include some partial interpretations in the HTA-definition, besides the broad and comprehensive one used today. PMID- 9739607 TI - [Genetic markers for stomach ulcer. A study of 3,387 men aged 54-74 years from The Copenhagen Male Study]. AB - BACKGROUND: Knowledge on the genetic risk of peptic ulcer has predominantly been based on hospital materials. To minimize selection bias, we tested the association between some genetic markers and the risk of peptic ulcer in a large scale epidemiologic design. MATERIAL AND METHODS: Some 3,387 white men aged 54-74 years were investigated and reported their history of peptic ulcer. Information about hospitalization and operation was collected from registers. RESULTS: The lifetime prevalence of peptic ulcer in men with Lewis phenotype Le(a + b-) and non-secretors of ABH antigen was 15%, significantly higher than others, 11%, p; the risk in phenotypes O and A were equally high, 12%, and among other ABO phenotypes, 7%, p < 0.05. Men with phenotype O had significantly higher risk of hospitalization than others, p < 0.01. The atributable risk of peptic ulcer in men who were Le(a + b-) or non-secretors, with O or A phenotypes, was 37%. No association was found with with complement C3, MNS blood group, or Rhesus blood group phenotypes. CONCLUSIONS: 1) The Le(a + b-) phenotype and the ABH non secretor trait are relevant genetic markers of peptic ulcer. We suggest that these men have an increased subceptibility to Helicobacter pylori infection. 2) This study challenges the importance of the ABO blood group: lifetime prevalence was equally high among men with O and A phenotype, with more severe cases in men with phenotype O. PMID- 9739608 TI - [Planning and specification of activities at a department of surgery]. PMID- 9739609 TI - [Local registration problems in connection with data collection at an emergency department]. PMID- 9739610 TI - [Parkinson disease. Alpha-synuclein is the first molecular help]. PMID- 9739611 TI - [Medical humor--no ghost. The Nordic Society for Medical Humor will be established in 1999]. PMID- 9739612 TI - [On the report of SSVF on differences in the handling of research funds]. PMID- 9739613 TI - [Iron deficiency in children during the first year of life]. PMID- 9739614 TI - [Babesiosis]. PMID- 9739615 TI - [A rapid determination of allantoin by high-performance liquid chromatography using tris(hydroxymethyl)aminomethane-HCl buffer as a mobile phase]. AB - A rapid and simple method for the determination of allantoin in pharmaceuticals by reversed-phase ion-pair high-performance liquid chromatography using an ODS column was presented. In general, it is difficult to retain allantoin to the ODS column owing to its very low hydrophobicity. We solved these problems by the use of a Tris-HCl buffer (pH 7.5) containing tetra-n-hexyl-ammonium bromide (THAB) as an ion-pair reagent for the mobile phase. Comparatively low concentrations of Tris-HCl buffer (0.9 mM) and THAB (0.5 mM) gave a high capacity factor (k'). As a results of the examination of the chromatographic behavior, it is confirmed that the retention mechanism of allantoin to the ODS column on the present method was not the ion-pair mode, but the ion-exchange mode. Calibration curves for allantoin showed a good linearity in the range of 10 to 400 micrograms/ml (r = 0.9999). The reproducibility (R.S.D., n = 6) was invariably good (0.37%). The lowest concentration of allantoin for the determination was 200 ng per 20 microliters of injection. The present method was successfully applied to the determination of allantoin in commercial eyedrops with good recovery (99.4%). It was found that allantoin in pharmaceuticals could be determined by the present method in short time and without any complicated derivatization. PMID- 9739616 TI - [Effect of carbon tetrachloride-induced hepatic injury on stereoselective N demethylation of chlorpheniramine by rat hepatic cytochrome P450 2C11 isozyme]. AB - We examined the effect of a carbon tetrachloride (CCl4)-induced hepatic injury on the stereoselective N-demethylation of RS-(+/-)-chlorpheniramine (Chp) by cytochrome P450 (CYP) 2C11 isozyme. In the non-treated rat liver microsomes, the stereoselective N-demethylation of racemic Chp was observed. However, in the CCl4 treated (0.5 ml/kg, i.p.) rat liver microsomes, the N-demethylation activities of S-(+)- and R-(-)-Chp decreased continuously up to the third day after the treatment with CCl4, and reached about 9 and 13% of control values, respectively, and the stereoselective N-demethylation of Chp was not observed. Moreover, in the liver microsomes at the 7th day after the treatment with CCl4, the N demethylation activities of both enantiomers recovered to an original level, and the stereoselective N-demethylation of Chp was again observed. The addition of 30 microliters of the anti-rat CYP2C11 serum to the reaction mixture containing 1 mg of microsomal protein inhibited the formation of monodesmethylchlorpheniramine (DMChp) from both enantiomers to 74 and 57% of the control values for S-(+)- and R-(-)-Chp, respectively. In the liver microsomes of a male rat at the 1st day after the treatment of CCl4, the addition of the anti-rat CYP 2C11 serum (30 microliters) also caused 25% inhibition of the formation of DMChp from S-(+)-Chp, but anti-rat CYP2C11 had no inhibitory effect on the rates of microsomal N demethylation of R-(-)-enantiomer. On the other hand, in the liver microsomes of a male rat at the 7th day after the treatment with CCl4, the anti-rat CYP2C11 serum had an inhibitory effect on the rates of microsomal N-demethylation of either S-(+)- or R-(-)-enantiomers again. Moreover, it was confirmed by Western blotting analysis that the density of the stained bands of CYP2C11 in the liver microsomes from male rats at the 1st, 2nd and 3rd days after the treatment with CCl4, was thinner than that from non-treatment male rats. These results indicated that the changes of N-demethylation activities of Chp in the CCl4-induced hepatic injury were due to the variation of microsomal CYP2C11. PMID- 9739617 TI - [Analysis of contact dermatitis by cytokines. III.--Sensitization and crossreaction of phenolic compounds]. AB - We examined sensitization and crossreaction by the guinea pig maximization test (GPMT) with phenolic compounds. Skin specimens were collected from earlobes of BALB/c mice on 24 h after elicitation phase of contact hypersensitivity reaction (CHR) with phenolic compounds. The expression of cytokines of skin specimens was examined by the reverse transcriptase/polymerase chain reaction (RT-PCR) method. Consequently, phenolic compounds which showed positive reaction in GPMT expressed IL-2 and IFN-gamma on 24 h after elicitation, and some phenolic compounds showed marked crossreaction. Therefore, it was found that on several phenolic compounds, dimerization of these compounds from monomer to dimer decrease sensitization. PMID- 9739618 TI - Continuous and discrete mathematical models of tumor-induced angiogenesis. AB - Angiogenesis, the formation of blood vessels from a pre-existing vasculature, is a process whereby capillary sprouts are formed in response to externally supplied chemical stimuli. The sprouts then grow and develop, driven initially by endothelial-cell migration, and organize themselves into a dendritic structure. Subsequent cell proliferation near the sprout tip permits further extension of the capillary and ultimately completes the process. Angiogenesis occurs during embryogenesis, wound healing, arthritis and during the growth of solid tumors. In this paper we present both continuous and discrete mathematical models which describe the formation of the capillary sprout network in response to chemical stimuli (tumor angiogenic factors, TAF) supplied by a solid tumor. The models also take into account essential endothelial cell-extracellular matrix interactions via the inclusion of the matrix macromolecule fibronectin. The continuous model consists of a system of nonlinear partial differential equations describing the initial migratory response of endothelial cells to the TAF and the fibronectin. Numerical simulations of the system, using parameter values based on experimental data, are presented and compared qualitatively with in vivo experiments. We then use a discretized form of the partial differential equations to develop a biased random-walk model which enables us to track individual endothelial cells at the sprout tips and incorporate anastomosis, mitosis and branching explicitly into the model. The theoretical capillary networks generated by computer simulations of the discrete model are compared with the morphology of capillary networks observed in in vivo experiments. PMID- 9739619 TI - Restricted-range gradients and travelling fronts in a model of juxtacrine cell relay. AB - Patterning events in development often depend on the transmission over a range of several cell diameters of signals emanating from a localized source. Experimental studies of such long-range signalling by members of the TGF-beta family of growth factors suggests that a cell-relay mechanism in which cells signal only with their immediate neighbours (i.e., juxtacrine signalling) may be operating in some tissues. Here, this possibility is investigated through the analysis of a model of juxtacrine signalling. Depending on the strength of the signal relay between cells, a localized signal source can generate either stable gradients or travelling fronts of cell activation. Both of these behaviors could in principle be involved in the long-range transmission of signals and patterning of cell fates by cell relays. There are significant and surprising differences between the gradients generated by the mechanism studied here, and those generated by the diffusion of a morphogen. In particular, there is an upper limit on the distance over which any given level of cell activation can be attained in a relay-mediated gradient. irrespective of the strength of signal source. PMID- 9739620 TI - Wavelet-based analysis of human blood-flow dynamics. AB - To analyze signals measured from human blood flow in the time-frequency domain, we used the wavelet transform which gives good time resolution for high-frequency components and good frequency resolution for low-frequency components. Five characteristic frequency peaks, corresponding to five almost periodic rhythmic activities, were found on the time scale of minutes. These oscillations were characterized by time and spatial invariant measures. The potential of this approach in studying the blood-flow dynamics was illustrated by revealing differences between the groups of control subjects and athletes. PMID- 9739621 TI - Distribution of attachment events relative to actin binding sites as evidenced in a bidirectional actomyosin interaction model. AB - Optical trapping is one of the most evolving technologies that measures biophysical quantities and provides insights into some of the fundamental questions in the study of molecular motor proteins such as myosin. Several laboratories have successfully used this technique to observe and score nanometre size displacements produced by myosin on interacting with actin. We have studied the distribution of attachment events for two myosin molecules with different orientations interacting with an actin filament within the framework of a Langevin-type bidirectional mathematical model. When myosin is detached from actin, our model predicts Brownian displacements centred at 0 +/- 8 nm (mean +/- SD, n = 251,058). When attached, the time-averaged displacements of the actin filament system produced step sizes with peaks of 8 +/- 6 nm (mean +/- SD, n = 22,174) (forward displacements) and -8 +/- 6 nm (mean +/- SD, n = 26,769) (reverse displacements). We infer from our results that the population distribution of attachment events is strongly dependent on (i) the magnitude of the Brownian displacements, (ii) the location of the actin binding sites relative to the myosin molecules, (iii) the orientation of the myosin molecules, and (iv) the relative kinetics (rate constants) for the forward and reverse displacement events. PMID- 9739622 TI - Cyclopeptide alkaloids. PMID- 9739624 TI - Managing the uncertainties of low-level radioactive waste disposal. AB - The disposal of low-level radioactive waste (LLRW) entails financial and safety risks not common to most market commodities. This manifests debilitating uncertainty regarding future waste volume and disposal technology performance in the market for waste disposal services. Dealing with the publicly perceived risks of LLRW disposal increases the total cost of the technology by an order of magnitude, relative to traditional shallow land burial. Therefore, this analysis first examines five proposed disposal facility designs and quantifies the costs associated with these two important sources of uncertainty. Based upon this analysis, a marketable disposal permit mechanism is proposed and analyzed for the purpose of reducing market uncertainty and thereby facilitating a market solution to the waste disposal problem. In addition to quantifying the costs, the results illustrate the ways in which the design of a technology is influenced by its institutional environment, and vice versa. PMID- 9739623 TI - The association between ambient carbon monoxide levels and daily mortality in Toronto, Canada. AB - The role of ambient levels of carbon monoxide (CO) in the exacerbation of heart problems in individuals with both cardiac and other diseases was examined by comparing daily variations in CO levels and daily fluctuations in nonaccidental mortality in metropolitan Toronto for the 15-year period 1980-1994. After adjusting the mortality time series for day-of-the-week effects, nonparametic smoothed functions of day of study and weather variables, statistically significant positive associations were observed between daily fluctuations in mortality and ambient levels of carbon monoxide, nitrogen dioxide, sulfur dioxide, coefficient of haze, total suspended particulate matter, sulfates, and estimated PM2.5 and PM10. However, the effects of this complex mixture of air pollutants could be almost completely explained by the levels of CO and total suspended particulates (TSP). Of the 40 daily nonaccidental deaths in metropolitan Toronto, 4.7% (95% confidence interval of 3.4%-6.1%) could be attributable to CO while TSP contributed an additional 1.0% (95% confidence interval of 0.2-1.9%), based on changes in CO and TSP equivalent to their average concentrations. Statistically significant positive associations were observed between CO and mortality in all seasons, age, and disease groupings examined. Carbon monoxide should be considered as a potential public health risk to urban populations at current ambient exposure levels. PMID- 9739625 TI - Evaluation of tire-derived fuel for use in nitrogen oxide reduction by reburning. AB - Tire-derived fuel (TDF) was tested in a small-scale (44 kW or 150,000 Btu/hr) combustor to determine its feasibility as a fuel for use in reburning for control of nitrogen oxide (NO). TDF was gravity-fed into upward flowing combustion gases from a primary natural gas flame doped with ammonia to simulate a high NO combustion process. Emissions of NO, oxygen, carbon dioxide, carbon monoxide, and particulate matter were measured. The tests varied the nominal primary NO level from 600 to 1,200 ppm and the primary stoichiometry from 1.1 to 1.2, and used both natural gas and TDF as reburn fuels. The reburn injection rate was varied to achieve 8-20% of the total heat input from the reburn fuel. NO emissions reductions ranged between 20 and 63% when using TDF, depending upon the rate of TDF injection, primary NO, and primary stoichiometry. NO emission reductions when using natural gas as the reburn fuel were consistently higher than those when using TDF. While additional work remains to optimize the process and evaluate costs, TDF has been shown to have the potential to be a technically viable reburning fuel. PMID- 9739626 TI - The Boston residential nitrogen dioxide characterization study: classification and prediction of indoor NO2 exposure. AB - Many epidemiological studies have used house characteristics associated with indoor sources as simplified proxies for personal nitrogen dioxide (NO2) exposure. Stove type and presence of a pilot light often been used as the two key characteristics, but significant overlaps have remained in the NO2 concentrations in the exposed and unexposed groups. This has contributed to inconsistencies in epidemiological findings, due to potential misclassification of exposure. In this study, other possible proxies were analyzed by cross-table analyses and were investigated in terms of improvements in both classification and predictive power. Adding building type to the above two proxies resulted in 0-5% of households with concentrations overlapping the observed range for the opposing stratum, compared with 22-42% for the two-proxy model. In spite of this performance, the predictive power of regression models for indoor NO2 was not improved by the addition of the third proxy, and the potential sample population was significantly limited. Using these analytical methods to choose descriptive proxies and evaluate the tradeoffs in their implementation can help epidemiological studies improve their designs and therefore optimize the robustness of their conclusions. PMID- 9739627 TI - The relationship between the quantity of heavy metal and PAHs in fly ash. AB - Heavy metal and polycyclic aromatic hydrocarbons (PAHs) in flue gas have received considerable attention in recent years due to their mutagenic or carcinogenic properties. The present study is carried out to investigate the influence of the quantity of heavy metals on PAHs formation in fly ash. A fluidized bed incinerator was used in this experiment to obtain fly ash of chemical similarity by incinerating various compositions of waste. The obtained fly ash, both with and without heavy metal, were used to adsorb the PAHs in the flue gas and to investigate the formation of PAHs in fly ash. The results indicate that carbon and heavy metals most greatly influence the formation of PAHs in the fly ash. Carbon is absorptive; heavy metals encourage not only absorption of PAHs but also catalyze PAHs formation. PMID- 9739628 TI - Kinetic sequential sampling (KSS) system: an automated sampling system for measuring vertical concentration profiles of airborne particles. AB - An electronically controlled lift system carrying a real-time particle monitor has been developed for sampling air sequentially, at different heights within the breathing zone. Data are automatically logged at the different receptor levels, for the determination of average vertical concentration profiles of airborne particulate matter. The system is easy to operate, portable, and easily extended to different heights or modified for use with other types of monitors (e.g., a portable CO analyzer). For measuring airborne particle concentrations, a Grimm Dust Monitor 1.104/5 was used. The results of trial runs, which were carried out indoors and in a relatively open semi-rural area, are presented, and applications of the kinetic sequential sampling (KSS) system are discussed. PMID- 9739629 TI - Perceptions of empathy and client satisfaction with managed behavioral health care. AB - Managed care continues to revolutionize the provision of mental health services in the United States. Long-term, open-ended therapies have been replaced by short term, highly focused interventions. Increasingly, managed care organizations rely on standardized preferred practice guidelines to give direction and focus to social work and other therapeutic interventions. Critics argue that changes effected by managed care, particularly the use of treatment guidelines, depersonalize the client-worker relationship and significantly reduce the role of empathy in the therapeutic process. Moreover, these critics suggest that overall client satisfaction with mental health services has deteriorated. This article presents a study that examined clients' perceptions of empathy and overall satisfaction with managed behavioral health care when the clients were in unstructured individual therapy or in time-limited standardized group therapy. The results reveal no significant difference in the clients' perception of empathy or of their overall satisfaction regardless of the type of treatment they received. This article describes the rationale and design of the study, presents the results, and discusses the implications for social work practice. PMID- 9739631 TI - Recovered memory therapy: a dubious practice technique. AB - This article examines the validity of memory work as well as the evidence for the efficacy of therapeutic interventions based in the recovery of childhood sexual abuse memories. Evidence suggests that both true and false memories can be recovered using memory work techniques, and there is no evidence that reliable discriminations can be made between them. Similarly, there is no empirical evidence to suggest that recovered memory therapy results in improved outcomes for participating clients. The article reviews current treatment outcome research and suggests that participation in recovered memory therapy may be harmful to clients. PMID- 9739630 TI - Outdated practitioner views about family culpability and severe mental disorders. AB - Current theories and research about the etiology and treatment of psychotic disorders increasingly point to the importance of biological factors. Accompanying this shift in the etiological literature has been an accumulation of evidence indicating the need to move away from treatment modalities that make families of people with psychotic disorders feel culpable in the causation or perpetuation of their relatives' disorders. The current study reports the development of a reliable and valid scale to assess the extent to which practitioners have made this shift. It also reports two surveys, the findings of which imply grounds for concern about what many practitioners do when working with clients with severe and persistent mental illness and their families. PMID- 9739632 TI - People matter: client-reported interpersonal interaction and its impact on symptoms of schizophrenia. AB - The current emphasis of biological research on schizophrenia has overlooked the importance of the environment in understanding fluctuations in symptoms of this devastating mental illness. One way of reconnecting the individual with his or her environment is by better understanding the subjective experience of the illness and what environmental factors lead to changes in the symptoms of schizophrenia. This study examines the nature of the subjective experience of schizophrenia during two symptom fluctuation episodes, what individuals perceived as the most important symptom fluctuations they experienced during those episodes, and what individuals perceived as causing the fluctuations in symptoms. During both episodes individuals reported primarily mood/affect and physical symptom changes and identified interpersonal interactions as the primary cause of the symptom changes. Mental health professionals should be more aware of and better assess the role of interpersonal interaction as it relates to changes and decompensation in people with schizophrenia. PMID- 9739633 TI - Associations among economic need, self-esteem, and Israeli Arab women's attitudes toward and use of professional services. AB - This study examines the effects of economic need and self-esteem on the attitudes toward and use of professional (instrumental and psychotherapeutic) services by Arab women living in the mixed Arab-Jewish town of Jaffa, Israel. Findings show that self-esteem was associated with the women's help-seeking behavior but not their attitudes only when economic need was not taken into account. When economic need was included in the analyses, the effect of self-esteem disappeared altogether. These findings point to the importance of economic need in actual help seeking and cast doubt on the adequacy of the "threat to self-esteem" model to explain underutilization of professional services. PMID- 9739634 TI - Improving compliance with the initial outpatient session among discharged inpatient dual diagnosis clients. PMID- 9739635 TI - The power of PowerPoint. PMID- 9739636 TI - Children and stress. PMID- 9739637 TI - Pediatrics without walls. PMID- 9739638 TI - Support groups: an empowering, experiential strategy. AB - The authors describe a student-facilitated support group experience initiated at student request and designed for RN-BSN students. Students report they emerged enlightened about group theory, empowered to share their knowledge of groups, and energized to initiate groups in their work settings. If educators make the learning experience safe, practice letting go and being vigilant, and celebrate group successes, students learn how to initiate, facilitate, and terminate small groups. PMID- 9739639 TI - Community clinical resources: an assessment tool. PMID- 9739640 TI - Introducing ADN students to nursing research. AB - Every nurse, regardless of educational preparation, should be involved in and benefit from nursing research. The research process needs to become an integral part of nursing practice. In this article, the authors emphasize the importance of nursing research in the associate degree nursing curriculum, emphasizing strategies that enable the ADN graduate to appreciate research reports and use the knowledge in the clinical practice setting. PMID- 9739641 TI - The clinical experience of novice students in nursing. AB - The learning experience of nursing students in their first clinical laboratory in a hospital was examined in a qualitative investigation. Graduate students in a nursing research seminar course participated as co-investigators in the study of clinical learning among sophomore nursing students. Findings revealed that sophomore students in nursing reflected on their role in the clinical setting and in nursing; pursued ways to learn in clinical settings; actively sought mentors; made connections to staff, patients, and peers; and searched for ways to validate the competence of their beginning skills. Parallels of the students' behaviors to the novice-to-expert paradigm were found. The study was valuable for both undergraduate and graduate students involved in the investigation. PMID- 9739642 TI - Home care and rehabilitation nursing: a winning combination. PMID- 9739643 TI - Computerized testing in a nursing curriculum. A case study. AB - Computerized testing has a place in nursing curricula with benefits shared between faculty and students. The author chronicles the activities surrounding the implementation of computerized testing in a nursing program. Key issues concerning faculty involve software selection, test proctoring, student honesty, and security of computer files. Students prefer the new testing format, question feedback and the option of Saturday examinations and they feel more prepared for the computerized licensure exam. PMID- 9739644 TI - Infusing teaching thinking skills into subject-area instruction. AB - Students can be taught clinical reasoning actively with the adaptation of the infusion approach for "teaching of" thinking. The authors demonstrate the "teaching of" critical thinking strategies for diagnostic decisions. The instructor uses a thinking map and models the process of diagnosis step by step. Working in small groups, students practice, monitor, and revise their strategies of clinical reasoning. PMID- 9739645 TI - Clinical correlation map. A tool for linking theory and practice. AB - To facilitate application of theoretical knowledge to clinical practice, students develop a clinical correlation map that diagrams the relationships between each client's assessment and diagnostic data, medical treatment, human responses, and nursing diagnoses. In addition to the tool's benefits to the student, faculty can quickly assess the student's understanding of pathophysiology, diagnostic testing, treatment, nursing diagnoses, and interventions. PMID- 9739646 TI - Development of a faculty work load formula. The teaching component. AB - In response to a university mandate and general faculty dissatisfaction with work assignments, faculty developed a faculty work load formula. The authors discuss the development of the teaching portion of the formula. Implementation of the formula and publication of teaching work loads has resulted in greater awareness of other's responsibilities, improved faculty morale, increased accountability, and a greater sense of control. Some problem areas are also discussed. PMID- 9739647 TI - Serendipity. A focus group turned hermeneutic evaluation. AB - When a focus group exploring the influence of feminism on students' lives became a hermeneutic evaluation, we understood students' lived experience, gained a retrospective view of students' developing feminist voice, and learned how feminist education influenced students' personal and professional lives. Nurse educators in programs espousing phenomenological, practice-driven, dialogic education are encouraged to integrate hermeneutic evaluation strategies so faculty-student groups can make courses and curricula relevant and meaningful. PMID- 9739648 TI - Faculty practice. Why and how. AB - The decision to integrate a planned program of faculty practice into a school of nursing requires careful analysis and consideration. The challenge is not only "why should we do it?" but "how should we do it?". The author highlights university, school of nursing, and faculty issues and concerns that relate to implementation of a faculty practice plan. PMID- 9739649 TI - "Doctors" leaders swing to support NHS reforms. PMID- 9739650 TI - New story, but old mistakes. PMID- 9739651 TI - Trust saves money by taking students' homes. PMID- 9739652 TI - Role reversal. PMID- 9739653 TI - Fit for a king. PMID- 9739654 TI - Firing line. PMID- 9739655 TI - The Christine Hancock column. PMID- 9739656 TI - Clinical placements. PMID- 9739657 TI - Places please. PMID- 9739658 TI - Women's health. PMID- 9739659 TI - Writing partners: the pros and cons of co-authorship. PMID- 9739660 TI - Capacity building in the new South Africa: contribution of nursing research. AB - In this article, the author identifies the contribution of nursing research to building the capacity of healthcare services to care for the people of the Republic of South Africa. The author specifies the areas where nursing research can make a significant contribution and makes recommendations for enhancing nursing research in the emerging republic. PMID- 9739661 TI - Inequalities in health: effects of socio-economic status. PMID- 9739662 TI - Clinical supervision: characteristics of a good supervisor. AB - In this review of the literature, the author evaluates research into supervisor and supervisee perceptions of the attributes of good clinical supervisors. Nursing studies are reviewed but, because of the relative dearth of studies on the topic, the author draws on the counselling, psychology and psychotherapy literature to develop comparisons. PMID- 9739663 TI - Dermatology. PMID- 9739664 TI - Last Offices--2. PMID- 9739665 TI - Wake up to nurse power. PMID- 9739666 TI - Health guide for older women unveiled. PMID- 9739667 TI - Invisible women. PMID- 9739668 TI - The tragic roundabout. PMID- 9739669 TI - One of a kind. PMID- 9739670 TI - In the dock. PMID- 9739671 TI - Who will be left holding the baby? PMID- 9739672 TI - District nurses need to shout to make people listen. PMID- 9739673 TI - A plethora of accents. PMID- 9739674 TI - Fifty years on from its inception. PMID- 9739675 TI - All you need to know about clinical effectiveness. PMID- 9739676 TI - Do the right thing. PMID- 9739677 TI - Photography and nursing do mix. PMID- 9739678 TI - Warts up, nurse? PMID- 9739679 TI - Culture shock. AB - Parasathi Teare argues that attempts to eradicate female circumcision are misguided. Instead, we should demand effective medical care for women in developing countries. PMID- 9739680 TI - The pain of rejection. PMID- 9739681 TI - At the cutting edge. PMID- 9739682 TI - A harmful procedure. PMID- 9739683 TI - Only scratching the surface. PMID- 9739684 TI - It's a family affair. Interview by Adam Legge. PMID- 9739685 TI - Nursing the nomads. PMID- 9739686 TI - Patients who want to complain about their medical treatment can find themselves lost in a bureaucratic maze. PMID- 9739687 TI - Nurse-led primary care act pilot schemes: threat or opportunity? PMID- 9739688 TI - Is audit a task for nurses? AB - Psychiatric nurses in the residential sector of St George's Hospital in Morpeth, Northumberland, worked with a quality coordinator to devise a means of auditing the quality of care in their wards. Their major objective was to bring the process to the grass roots of nursing care and involve front-line staff in designing and carrying it out. PMID- 9739689 TI - Using video for clients with severe learning disabilities. PMID- 9739690 TI - Partnership encourages patients to comply with treatment. PMID- 9739691 TI - Preparing patients for endoscopy. PMID- 9739692 TI - Growing up with a stoma. PMID- 9739693 TI - Frankly underwhelming. PMID- 9739695 TI - Nursing a grievance. PMID- 9739697 TI - 20 questions. Interview by Charles Webster. PMID- 9739694 TI - Last among equals. PMID- 9739698 TI - The future is yours. PMID- 9739699 TI - The long and winding road. PMID- 9739701 TI - Last offices--1. PMID- 9739700 TI - Wide awake and stretching. PMID- 9739702 TI - In Bevan's name. PMID- 9739704 TI - Partners in care. PMID- 9739705 TI - Face to face. Interview by Eileen Fursland. PMID- 9739703 TI - Looking after number one. Interview by Esther Leach. PMID- 9739706 TI - All our yesterdays. Interview by Eileen Fursland. PMID- 9739708 TI - Sister Plume. PMID- 9739707 TI - The golden girls. Interview by Eileen Fursland. PMID- 9739709 TI - On the continental shelf. PMID- 9739710 TI - One nurse is sick of care homes that smell of incontinence. PMID- 9739711 TI - Nursing supply and demand: reviewing the evidence. AB - This paper reviews research on the supply of, and demand for nurse. It examines the utility of various data indicators of nursing shortages and highlights potential shortcomings in their application. The paper also assesses the links between nurses' pay and nurses' labour market behaviour, reviewing the comparatively few papers that have taken a UK perspective on this issue. The current mismatch between nurses' supply and demand in the UK is also examined, with the clear indication that there is potential for an increase in mismatch as demand increases, while future supply appears problematic. PMID- 9739712 TI - Sickle cell disease and the carer-client relationship. PMID- 9739713 TI - Safe patient transportation: nurses can make a difference. PMID- 9739714 TI - Culturally sensitive care in a forensic setting. AB - In the forensic service there is a disproportionate number of ethnic minority patients. Providing culturally sensitive care has not been given a high priority within formal nurse education in the past. This article describes a training programme for nurses working in a forensic service aimed at promoting culturally sensitive care. The programme included methods of raising awareness, motivation and interest in training on this subject. PMID- 9739715 TI - Giving contraception advice. PMID- 9739716 TI - [Medical strategy. Care at the end of life]. PMID- 9739717 TI - [A nurse's point of view. The nurse's experience regarding death]. PMID- 9739718 TI - ["One is used to it ... one never gets used to it"]. PMID- 9739719 TI - [The implementation of controlled analgesia]. PMID- 9739720 TI - [Nursing care. Grief of the families]. PMID- 9739721 TI - [Collaboration between nurses and volunteers in the home]. PMID- 9739722 TI - [Nursing care. Common nursing records and work place]. PMID- 9739723 TI - [Putting intensity into the caring relationship]. PMID- 9739724 TI - [Palliative care: between uncertainty and equilibrium]. PMID- 9739725 TI - [Abortion and guilt]. PMID- 9739726 TI - [How to use the Saint Louis mouthwash]. PMID- 9739727 TI - [The role of the state and of health insurance]. PMID- 9739728 TI - [An inquiry about satisfaction of the patients at the Hyeres Health Center]. PMID- 9739729 TI - [Nursing research at the AP-HP]. PMID- 9739730 TI - [The metabolism of drugs]. PMID- 9739731 TI - [How to deal with respiratory distress]. PMID- 9739732 TI - [Pain management]. PMID- 9739733 TI - [Gastro-duodenal ulcer. Essential clinical facts]. PMID- 9739734 TI - Annual meeting of the American Society of Anesthesiologists. Orlando, Florida, USA. October 17-21, 1998. Abstracts. PMID- 9739735 TI - American College of Emergency Physicians Research Forum. San Diego, California, USA. October 11-12, 1998. Abstracts. PMID- 9739736 TI - IX. International Congress on Neuromuscular Diseases. Adelaide, Australia, August 30-September 4, 1998. Abstracts. PMID- 9739737 TI - American Academy of Pediatrics 1998 annual meeting. San Francisco, California, USA. October 16-21, 1998. Abstracts. PMID- 9739738 TI - Through the eyes of pop culture. PMID- 9739739 TI - HHS calls for inclusion of pregnant women. PMID- 9739741 TI - Teaching patients how to use PCA. PMID- 9739740 TI - Anticipating drug response. A commonly used anticoalgulant has many drug-drug interactions. PMID- 9739742 TI - Delegation revisited. PMID- 9739743 TI - Coping with chronic illness. Nursing knowledge and compassion can empower ill or disabled teens. PMID- 9739744 TI - The latex threat. PMID- 9739745 TI - Recollections of an expatriate nurse. PMID- 9739746 TI - Cultural competence: a nursing dialogue 2. PMID- 9739747 TI - Gastroesophageal reflux disease. PMID- 9739748 TI - No heavy lifting. Making safety work. PMID- 9739749 TI - Emergency! Stab wound. PMID- 9739750 TI - The dying game. PMID- 9739751 TI - Migrant health: a harvest of poverty. PMID- 9739752 TI - When long-term residents require emergent care. PMID- 9739753 TI - Pain management needs of patients with HIV or AIDS who also have a history of chemical dependency. PMID- 9739754 TI - Male nurses still face bias. PMID- 9739755 TI - Creutzfeldt-Jakob disease. PMID- 9739757 TI - Nurses' voices needed in halls of Congress. PMID- 9739756 TI - Nursing in the field. Mobile health units are an important part of bringing health care to communities. PMID- 9739758 TI - The papillomavirus E6 proteins. AB - Specific types of human papillomaviruses (HPV) are strongly associated with the development of cervical cancer. The E6 gene from cancer-related HPVs has exhibited functions in tumorigenesis, regulation of transcription, telomerase, and apoptosis. Cancer-related HPVs E6 proteins bind the tumor suppressor p53 and promotes its degradation through an ubiquitin-dependent pathway. Several additional cellular E6-binding proteins have recently been identified and implicated in playing roles in p53-independent functions of E6. PMID- 9739759 TI - Progress in understanding the molecular pathogenesis of human lung cancer. AB - We review the molecular pathogenesis of lung cancer including alterations in dominant oncogenes, recessive oncogenes/tumor suppressor genes, alterations in growth regulatory signaling pathways, abnormalities in other pathways, such as apoptosis, autocrine and paracrine growth stimulatory loops, angiogenesis, and host immune responses, other mechanisms of genetic changes, such as microsatellite and methylation alterations, and the potential for inherited predisposition to lung cancer. These changes are related to multistage carcinogenesis involving preneoplastic lesions, and lung development and differentiation. The translational applications of these findings for developing new ways of early detection, prevention, treatment, and prognosis of lung cancer are discussed. PMID- 9739760 TI - The pS2/TFF1 trefoil factor, from basic research to clinical applications. AB - pS2/TFF1 trefoil factor is normally expressed in the stomach, and is found ectopically in gastrointestinal inflammatory disorders and in various carcinomas. It is involved in stomach ontogenesis and in the maintenance of the integrity of the mucosa, and may represent a pharmacological tool for prevention and healing of gastrointestinal ulcerations. In breast cancer, it can be used to select patients suitable for hormone therapy. pS2/TFF1 is a pleiotropic factor involved in mucin polymerization, cell motility, cell proliferation and/or differentiation, and possibly in the nervous system. PMID- 9739761 TI - Signal transduction via platelet-derived growth factor receptors. AB - Platelet-derived growth factor (PDGF) exerts its stimulatory effects on cell growth and motility by binding to two related protein tyrosine kinase receptors. Ligand binding induces receptor dimerization and autophosphorylation, allowing binding and activation of cytoplasmic SH2-domain containing signal transduction molecules. Thereby, a number of different signaling pathways are initiated leading to cell growth, actin reorganization migration and differentiation. Recent observations suggest that extensive cross-talk occurs between different signaling pathways, and that stimulatory signals are modulated by inhibitory signals arising in parallel. PMID- 9739762 TI - The use of chromatin templates to recreate transcriptional regulatory phenomena in vitro. PMID- 9739763 TI - [Suprasellar meningioma. A disease still frequently diagnosed too late]. AB - BACKGROUND AND OBJECTIVE: Suprasellar meningioma continues to be diagnosed very late after the onset of first eye symptoms. This study was aimed at demonstrating the effect of delay on the long-term visual loss. PATIENTS AND METHODS: In the course of a retrospective study all 53 consecutive patients operated on for suprasellar meningioma from 1982 to 1991 were contacted (47 women, 6 men; average age 49.5 years) 46 of the 49 surviving consented to the follow-up investigation. The extent of preoperative visual loss, tumour size, presence of optic nerve atrophy and duration of visual loss, data that provide an indirect measure of how soon the correct diagnosis was made, were analysed with regard to their effect on long-term ophthalmological results. RESULTS: The mean period elapsing from onset of first visual symptoms to the definitive diagnosis of suprasellar meningioma was 22.3 months. The data showed that the long-term results were the worse the later the diagnosis was made. CONCLUSIONS: The commonly very late diagnosis of suprasellar meningioma as cause of visual loss is an international problem and is presumably due to the low incidence of the tumour (1-2 cases per 1 mill. population per year). If long-term results are to be improved, primary care doctors must be made aware of the differential diagnosis of visual loss caused by pressure from a tumour. PMID- 9739764 TI - [Circumscribed apical left ventricular hypertrophy. Dynamic development and long term progression]. AB - HISTORY AND CLINICAL FINDINGS: A 64-year-old obese man had for 15 years suffered from exercise-independent retrosternal pressure sensation, radiating to the neck and back. Shortly after the onset of these symptoms he had undergone coronary angiography with negative results. But at that time the resting ECG showed discrete T wave negativity in the left precordial leads. INVESTIGATIONS: At the present admission the ECG showed deeply inverted T waves in the left precordial and limb leads and a positive Sokolow-Lyon index of 4.8 mV. Left ventricular angiography demonstrated in enddiastole a circumscribed myocardial hypertrophy limited to the apex and of typical "ace of spade" shape. DIAGNOSIS, TREATMENT AND COURSE: Left-heart catheterization and angiocardiography provided the diagnosis of circumscribed apical left ventricular hypertrophy (ALVH). As the patient had only minor symptoms no treatment was given. CONCLUSION: Circumscribed ALVH can show marked dynamic development in long-term observations. If there is marked T wave negativity, even with previously normal LV angiography, circumscribed ALVH should be included in the differential diagnosis. Patients with atypical angina pectoris and increasingly suggestive ECG changes should, even if previous coronary angiography had been negative, undergo transthoracic echocardiography with a high-frequency transducer, special attention being paid to muscular changes at the LV apex. PMID- 9739765 TI - [Subcutaneous manifestations of a centrocytic non-Hodgkin lymphoma at the injection site of a mistletoe preparation]. AB - HISTORY AND CLINICAL FINDINGS: A 73-year-old man, first diagnosed as having centrocytic Non-Hodgkin lymphoma 5 years previously, presented with subcutaneous nodes of the abdominal wall at precisely the sites of previous regular injections of a mistletoe preparation. These nodes had first appeared 5 weeks after the first injection. Injections were stopped and he reported to the out-patient clinic. Except for the visible and easily nodes in the anterior wall no other subcutaneous nodes were palpated. Prominent cervical lymphomas and swelling of the epicranial aponeurosis and lower lip had, according to the patient, been present for some time. INVESTIGATIONS: There was a T-lymphocytopenia, lactate dehydrogenase activity was raised to 255 U/l. The subcutaneous nodes in the anterior abdominal wall were also demonstrated by ultrasound. Computed tomography showed them as having the density of connective tissue and being up to 5 cm in diameter. The excised nodes histologically revealed to be infiltrations by the centrocytic lymphoma. TREATMENT AND COURSE: Once the mistletoe injections had been discontinued no further subcutaneous infiltrates were seen despite the progression of the lymphoma. Six weeks later the patient died of bilateral pneumonia. CONCLUSION: There are pointers that high concentrations of mistletoe preparations subcutaneously injected can have a growth-promoting action on cells of a centrocytic lymphoma. As part of a leukaemic "wash-out", these cells reach the subcutaneous tissue. This proliferative stimulus may have been mediated by a high local concentration of interleukin-6 liberated from the skin by mistletoe lectins. PMID- 9739766 TI - [T-cell response against viruses. A new non-cytotoxic antiviral strategy]. PMID- 9739767 TI - [Diagnosis of vertigo]. PMID- 9739768 TI - [Antioxidative vitamins: prevention of cardiovascular diseases]. PMID- 9739769 TI - [Recent developments in non-invasive heart imaging. Critical evaluation of the use of cardiac magnetic resonance tomography--results of an interdisciplinary workshop]. PMID- 9739770 TI - [Urine alcohol determination]. PMID- 9739771 TI - [Do relatives of Creutzfeldt-Jakob patients want to become genetically enlightened?]. PMID- 9739772 TI - Evaluation and risk stratification of patients with chest pain in the emergency department. Predictors of life-threatening events. AB - While assessing chest pain in the emergency department, physicians must first estimate the probability of acute ischemic states in the patient. This first estimate is based on the patient's history, physical examination, and electrocardiogram. Patients who meet the threshold for acute cardiac ischemia are further evaluated to confirm or exclude this diagnosis, while other life threatening factors are excluded. PMID- 9739773 TI - Early laboratory indicators of acute myocardial infarction. AB - Biochemical markers of myocardial injury have evolved so that the diagnosis or exclusion of acute myocardial infarction can be determined within a short time with a high degree of sensitivity and specificity. The use of these markers in patients complaining of chest pain allows for medically appropriate and cost effective triage decision making in the emergency department. PMID- 9739774 TI - Emergency management of acute myocardial infarction. Focus on pharmacologic therapy. AB - Treatment of acute myocardial infarction has evolved significantly in the past two decades. Reperfusion therapies of thrombolysis and percutaneous angioplasty are major advances that can be employed to save infarcting myocardium and reduce mortality. When reperfusion therapy is combined with the use of aspirin, beta blockade, heparin, and nitroglycerin, the emergency management of the patient with myocardial infarction can be completed. Outcomes in patients are determined by what happens in the first few minutes to hours after onset, and any delay in diagnosis or treatment may have significant consequences. This article reviews intervention and treatment strategies for acute myocardial infarction. PMID- 9739775 TI - Acute interventions for myocardial reperfusion. AB - The primary goal of treatment in acute myocardial infarction is reperfusion of the infarct-related artery in as short a time as possible. Present strategies for acute reperfusion include the use of thrombolytic agents and a variety of catheter-based interventions. This article presents a brief review of these strategies and discusses the patient subsets better served by a particular type of intervention. PMID- 9739776 TI - Arrhythmias associated with acute myocardial infarction and thrombolysis. AB - Ninety percent of patients with acute myocardial infarction have some cardiac rhythm abnormality, and approximately twenty-five percent have cardiac conduction disturbance within 24 hours following infarct onset. Almost any rhythm disturbance can be associated with acute myocardial infarction, including bradyarrhythmias, supraventricular tachyarrhythmias, ventricular arrhythmias, and atrioventricular block. With the advent of thrombolytic therapy, it was found that some rhythm disturbances in patients with acute myocardial infarction may be related to successful coronary artery reperfusion. This article addresses the role and treatment of arrhythmias and conduction disturbances that complicate the course of patients with acute infarction and thrombolysis. PMID- 9739777 TI - Evaluation of syncope in the emergency department. AB - Although most cases of syncope are benign, an adequate evaluation, which begins in the emergency department, is required to exclude life-threatening disorders. In addition, life-threatening disorders such as QT prolongation as well as confounding alternative diagnoses (e.g., seizure disorder) are also discussed. PMID- 9739778 TI - Syncope in children. AB - The sudden loss of consciousness in a child is concerning to both patients and their parents. Although most cases of syncope in children are benign, an adequate evaluation is required to exclude life-threatening disorders. Patient history and physical examination may be sufficient to define the cause of syncope in a large percentage of pediatric cases. The events and setting preceding the syncopal episode provide clues in defining the nature of the event. PMID- 9739779 TI - Ventricular arrhythmias in the elderly. AB - Sudden cardiac death (SCD) remains a significant medical problem in the United States. The incidence of SCD increases with advancing age because cardiovascular disease is more prevalent in the elderly. Management of ventricular arrhythmias in the elderly patient is especially challenging because of increased risk of interventional and pharmacologic therapies, altered pharmacokinetics of drugs, and sometimes unclear long-term benefits. PMID- 9739780 TI - Cardiopulmonary resuscitation in the elderly. Beneficial or an exercise in futility? AB - Sudden cardiac death is one of the leading causes of death and a major public health problem that particularly affects the elderly. Sudden cardiac death may be a terminal event after a prolonged debilitating and painful illness, or it may occur following many years of symptoms related to a cardiac disorder; however, in many elderly persons, the cardiac arrest may be the first manifestation of cardiac disease in a supposedly healthy and physically active person. Whether cardiopulmonary resuscitation should be performed in elderly patients who sustain cardiac arrest is a significant issue confronting the medical profession and the general public. Several questions must be answered when evaluating the decision of whether or not to perform cardiopulmonary resuscitation on an elderly patient. PMID- 9739781 TI - Inflammatory disorders of the heart. Pericarditis, myocarditis, and endocarditis. AB - The emergency physician frequently sees patients with cardiac complaints. Too often, ischemic heart disease is overwhelmingly considered to the exclusion of other diagnostic possibilities such as inflammatory cardiac disorders. These disorders can cause considerable discomfort, have long-term implications, or lead to more serious cardiac disorders. Emergency physicians must have a practical understanding of the diagnostic evaluation and therapeutic approach to patients with inflammatory cardiac disorders. PMID- 9739782 TI - Increased expression of cyclooxygenase-2 to -1 in human colorectal cancers and adenomas, but not in hyperplastic polyps. AB - BACKGROUND: Non-steroidal anti-inflammatory drugs can reduce the risk of colorectal cancer. Reportedly, mRNA expression of cyclooxygenase-2 (COX-2) is elevated in human colorectal cancers compared with accompanying normal mucosa. The present study was undertaken to establish a simple analytical procedure to quantify COX-2 expression levels and to characterize COX-2 expression levels in human colorectal cancers, adenomas and hyperplastic polyps. METHODS: The combination of PCR using common primers designed in the highly conserved regions and fluorescence-based single-strand conformation polymorphism (F-SSCP) analysis of the products is used for quantitative determination of the proportions of COX 2 mRNA in human colorectal cancers, adenomas, hyperplastic polyps and accompanying normal mucosa. RESULTS: The present F-SSCP analysis was a simple and powerful method for quantitative determination of the proportions of COX-2 mRNA. The proportion of COX-2 mRNA was higher in cancer tissues than in accompanying normal mucosa in 46 of the 50 cancers. There was no significant correlation between the increase of the COX-2 proportion and tumor location or stages. The enhanced COX-2 expression was also observed in colorectal adenomas. On the other hand, the proportion of COX-2 mRNA in hyperplastic polyps was not significantly different from that in normal mucosa. CONCLUSIONS: The proportion of COX-2 to COX 1 expression was elevated in most human colorectal cancers and adenomas, but not in hyperplastic polyps. Therefore, the increased proportion of COX-2 expression might be an early event in the carcinogenesis of colorectal cancer. PMID- 9739783 TI - Microsatellite instability is associated with the macroscopic configuration of neoplasms in patients with multiple colorectal adenomas. AB - BACKGROUND: The management of patients with multiple colorectal adenomas, 10-100 in number, is often troublesome clinically. To establish the reasonable management of such cases, genetic backgrounds should be made clear. METHODS: For a total of 19 adenomas and four carcinomas from four patients with multiple colorectal adenomas, we analyzed genetic instability at four selected microsatellite loci and screened exon 1-4 of the APC gene with special reference to macroscopic configurations of adenomas and carcinomas. RESULTS: RER-positive phenotypes were detected in none of 13 protruded type adenomas, two (33%) out of six superficial elevated type adenomas and two (50%) out of four carcinomas. We detected no mutation of exon 1-4 of the APC gene in any sample. In patients with multiple colorectral adenomas, the proportion of superficial elevated type adenomas exhibiting genetic instability was significantly higher than that of protruded type adenomas. CONCLUSIONS: The results suggested that RER-positive phenotype was an early event in tumorigenesis through superficial elevated type adenomas. PMID- 9739784 TI - Continuous infusion cisplatin and etoposide chemotherapy for cancer of unknown primary site (CUPS) in Taiwan, a region with a high prevalence of endemic viral infections. AB - BACKGROUND: To evaluate the efficacy and toxicity of cisplatin/etoposide continuous infusion chemotherapy for cancer of unknown primary site in Taiwan, a region with a high prevalence of endemic viral infections. METHOD: Between April 1994 and February 1996, 20 patients with a diagnosis of CUPS were treated, including 15 males and five females, of average age 63.3 years (range 41-83 years). Continuous intravenous infusion of etoposide 80 mg/m2 and cisplatin 25 mg/m2 was given for 3 days every 3 weeks. Pretreatment tumor marker and viral serology studies were performed for baseline evaluation. Nearly two-thirds of the patients had poorly differentiated carcinoma. The average number of metastatic sites was 2.65 (range 1-4), with liver and lymph node involvement predominating. RESULTS: The overall response rate was 25% (95% CI 17.7-32.3%); 30.7% for poorly differentiated cancers and 25% for well differentiated cancers. Median survival was 4 months (range 1-12 months), 4.8 months for patients attaining partial response. Toxicity was moderate, grade 3 and 4 neutropenia occurred in 55% and grade 3 and 4 thrombocytopenia in 40%; other toxicities were mild. CA125 and CA199 were elevated in more than 50% of patients. Viral serology studies were not significantly different from those of the indigenous population. CONCLUSION: Etoposide and cisplatin combination chemotherapy has modest activity in patients with extensive CUPS and, at the schedule and dosage given, it is associated with moderate toxicity. PMID- 9739785 TI - Interleukin-1 alpha producing synovial sarcoma with prolonged fever: a case report. AB - The patient had a synovial sarcoma of the monophasic fibrous type accompanied by prolonged spike fever. Wide excision of the tumor resulted in the disappearance of her fever. The tumor cells showed interleukin-1 alpha (IL-1 alpha) expression immunohistochemically. IL-1 alpha cDNA was detected in the tumor by RT-PCR, the sequences of which were identical with those of normal human IL-1 alpha molecules. These results indicated that IL-1 alpha, one of the fever-inducing cytokines, was expressed by sarcoma cells in the present case, which were thought to be a causative factor of the fever. PMID- 9739786 TI - A case of giant peritoneal loose bodies mimicking calcified leiomyoma originating from the rectum. AB - Two giant peritoneal loose bodies were found in the pelvis in a 79-year-old man. These bodies were demonstrated by computed tomography and magnetic resonance imaging to be well circumscribed masses and to have marked calcification in their central portion. Preoperatively, these bodies had been diagnosed as a calcified leiomyoma originating from the rectum; however, surgery revealed these lesions to be detached appendices epiploica. Histological examination showed that these peritoneal loose bodies consisted of thin layers of eosinophilic substance and had no cellular component. Small peritoneal loose bodies are occasionally found during laparotomy, but such large ones measuring 6 cm are very rare. In our case, accurate diagnosis could not be obtained preoperatively, because these loose bodies mimicked calcified leiomyoma of the rectum. PMID- 9739787 TI - Report of the Eleventh International Symposium of the Foundation for Promotion of Cancer Research: Basic and Clinical Research in Gastric Cancer. PMID- 9739788 TI - Cancer incidence in Japan in 1990: estimates based on data from Population-based Cancer Registries. The Research Group for Population-based Cancer Registration in Japan. PMID- 9739789 TI - A primer for genetic counseling of hereditary cancer--visits to the cancer hospitals in the United States. PMID- 9739790 TI - 'Breast cancer prevention trial'. PMID- 9739791 TI - NCI statement on animal studies of endostatin and angiostatin. PMID- 9739792 TI - The Harris-Benedict energy studies: additional considerations. PMID- 9739793 TI - Sounding the alarm for misuse of olestra-containing foods in binge-eating disorders. PMID- 9739794 TI - Healthy People 2010 offers ADA opportunity for action. PMID- 9739795 TI - Are phytoestrogens nature's cure for what ails us? A look at the research. Interview by Nancy I. Hahn. PMID- 9739796 TI - Patients report positive nutrition counseling outcomes. AB - OBJECTIVE: Assess outcomes of patient nutrition counseling. DESIGN: A descriptive study based on the results of a telephone interview performed 2 to 8 weeks after counseling. SUBJECTS/SETTING: Subjects were 400 adult patients referred for nutrition counseling at 2 academic health centers. Of these, 274 patients received nutrition counseling during hospitalization and 126 as outpatients. STATISTICAL ANALYSIS: Descriptive statistics were used to summarize data and the Mann-Whitney U statistic and logistic regressions were used to test significant differences (P < .05) between inpatient and outpatient counseling outcomes. RESULTS: Most patients (83%) gave a partial or full description of their diet modifications and 79% had a moderate or good understanding of their diet. Most patients reported that the dietitian's advice was suited to their special needs (88%) and that they knew what to eat (83%). A majority (62%) had made dietary changes, but 17% said they had had trouble changing their diets as suggested. After talking with a dietitian, 57% felt better emotionally, 37% felt better physically, 64% felt in control of their condition, and 43% noticed improved health indicators. Initial analysis indicated that outpatients reported better outcomes than inpatients; further analysis showed that these differences could be attributed to younger ages among the outpatient sample. APPLICATIONS/CONCLUSIONS: Patient nutrition counseling has positive outcomes. Therefore, key counseling points should be introduced or reinforced in inpatient settings, in conjunction with multiple-session protocols during the pre- and/or posthospitalization continuum of care. Dietitians, managers, administrators, and credentialing agencies should work together to secure and promote the necessary physical, personnel, and financial resources to make this happen. PMID- 9739797 TI - Estimation of individual intakes of folate in women of childbearing age with and without simulation of folic acid fortification. AB - OBJECTIVES: The objectives of this study were to examine variability of folate intake in order to estimate the number of days needed to accurately estimate intakes in women of childbearing age and to simulate the effect of folic acid fortification of cereals and grains on individual folate intake. DESIGN: Observational study of food intake over a 60-day period. SAMPLING: A convenience sample of 21 women completed food records on randomly assigned days within a 60 day period. OUTCOMES MEASURED: Folate intake and variance ratios of folate intake. STATISTICAL ANALYSIS: Repeated measures analysis of variance. RESULTS: Six days of food records were needed to describe folate intake of these women of childbearing age (18 to 45 years) with 20% attenuation of a correlation coefficient between dietary folate intake and another biological variable. Seven days of records were needed with simulated folic acid fortification (assuming fortification of 140 micrograms folic acid per 100 g flour) and 5 days were needed with supplements containing 200 to 400 micrograms folic acid in addition to folic acid fortification. Food folate intake was 288 +/- 195 micrograms; only 2 of the participants consumed the recommended 400 micrograms. With fortification, folate intake increased to 550 +/- 279 micrograms without supplements and 609 +/- 327 micrograms with supplements. APPLICATIONS: Individual intakes of folate should be assessed with at least 7 days of dietary records (20% attenuation). In this sample, when folic acid fortification was added to dietary intake, routine supplementation was not necessary to achieve folate intakes of 400 micrograms in the majority of participants. The practice of routine folic acid supplementation should be considered carefully to ensure that individual intakes of folate do not exceed the upper limits of safety. PMID- 9739798 TI - Using encoding and retrieval strategies to improve 24-hour dietary recalls among older adults. AB - OBJECTIVE: To examine whether using an encoding strategy and/or providing more support at the time of retrieval improves the accuracy of 24-hour dietary recalls among the elderly. DESIGN: Posttest-only control group design. SETTING: The sample was recruited through advertisements and at senior centers and a low income apartment building in rural central Pennsylvania. SUBJECTS: Study participants were 21 men and 73 women aged 58 years old and older. Everyone completed the study. INTERVENTION: The treatment group was unobtrusively guided in use of an encoding strategy before consuming the prepared meal. MAIN OUTCOME MEASURES: A 24-hour dietary recall and recognition tests were administered the next day for the foods consumed at the meal and for serving sizes of 5 of the foods. Memory tests were also administered. STATISTICAL ANALYSES PERFORMED: Linear regression was used to examine differences between the treatment and control groups and to identify variables that explained variation in the number of foods correctly recalled or recognized. The Chi 2 test was used to examine correct vs incorrect recall or recognition of the serving sizes of the 5 foods between the groups and to identify explanatory variables for this task. RESULTS: Subjects remembered more foods when they used an encoding strategy and when recognition replaced free recall; they performed best when both strategies were used. Use of this encoding strategy did not improve accurate recall or recognition of serving sizes of 5 foods; however, performances did improve when recognition replaced free recall. CONCLUSIONS: Among older adults, use of an encoding strategy and provision of support at the time of retrieval enhances memory of foods consumed but not of amounts consumed. To strengthen memory of foods consumed, older adults need to perform effortful memory tasks when they are eating. PMID- 9739799 TI - Guess who's cooking? The role of men in meal planning, shopping, and preparation in US families. AB - OBJECTIVES: To determine the role of men in meal-related tasks in households with both a male and female head, and to identify households in which the man is more likely to be involved in these tasks. DESIGN: Data collected as part of the US Department of Agriculture's 1994 Continuing Survey of Food Intakes of Individuals were analyzed. SUBJECTS/SETTING: All analyses were restricted to sampled persons who were identified as a male head of household residing in a household that also had a female head (N = 1,204). STATISTICAL ANALYSES: Frequency distributions were calculated and logistic regression analyses were conducted. RESULTS: Approximately 23%, 36%, and 27% of men reportedly were involved in meal planning, shopping, and preparation, respectively. Men in lower income and smaller households were more likely to be involved in each of the meal activities. Younger men and men in households in which the female head of household worked full-time were more likely to be involved in meal planning and preparation. IMPLICATIONS: Current education efforts to improve family nutrition tend to target the female rather than the male head of household. Our findings confirm that this focus is appropriate for most dual-headed households. PMID- 9739800 TI - Oxidative stress in critical care: is antioxidant supplementation beneficial? AB - Reactive oxygen species (ROS) are constantly produced in human beings under normal circumstances. Antioxidant systems help defend the body against ROS but may be overwhelmed during periods of oxidative stress, which can cause lipid peroxidation, damage to DNA, and cell death. Critical illness, such as sepsis or adult respiratory distress syndrome, can drastically increase the production of ROS and lead to oxidative stress. Sources of oxidative stress during critical illness include activation of the phagocytic cells of the immune system (the respiratory burst), the production of nitric oxide by the vascular endothelium, the release of iron and copper ions and metalloproteins, and the vascular damage caused by ischemia reperfusion. Only indirect measurements of ROS are available, but the presence of oxidative stress in critical illness is supported by clinical studies. In general, serum antioxidant vitamin concentrations seem to decrease and measures of oxidative stress seem to increase in critically ill populations. Oxidative stress has been associated with sepsis, shock, a need for mechanical ventilation, organ dysfunction, acute respiratory distress syndrome, disseminated intravascular coagulation, surgery, and the presence of an acute-phase response. In addition, higher levels of oxidative stress seem to occur in patients with more notable injuries. Dietary supplementation with antioxidant vitamins seems to be the logical answer to decreasing serum antioxidant concentrations, but antioxidants may have adverse effects. The benefit of supplementing antioxidants in critically ill populations has not been shown and requires further study. PMID- 9739801 TI - An artificial intelligence system for computer-assisted menu planning. AB - Planning nutritious and appetizing menus is a complex task that researchers have tried to computerize since the early 1960s. We have attempted to facilitate computer-assisted menu planning by modeling the reasoning an expert dietitian uses to plan menus. Two independent expert systems were built, each designed to plan a daily menu meeting the nutrition needs and personal preferences of an individual client. One system modeled rule-based, or logical, reasoning, whereas the other modeled case-based, or experiential, reasoning. The 2 systems were evaluated and their strengths and weaknesses identified. A hybrid system was built, combining the best of both systems. The hybrid system represents an important step forward because it plans daily menus in accordance with a person's needs and preferences; the Reference Daily Intakes; the Dietary Guidelines for Americans; and accepted aesthetic standards for color, texture, temperature, taste, and variety. Additional work to expand the system's scope and to enhance the user interface will be needed to make it a practical tool. Our system framework could be applied to special-purpose menu planning for patients in medical settings or adapted for institutional use. We conclude that an artificial intelligence approach has practical use for computer-assisted menu planning. PMID- 9739802 TI - Use of food quotients in human doubly labeled water studies: comparable results obtained with 4 widely used food intake methods. AB - Information on the macronutrient composition of the diet is needed in doubly labeled water studies to convert measured rates of carbon dioxide production into values for total energy expenditure. There is no general consensus, however, about the best method to determine food intake for this purpose. Four common methods of measuring food intake (7-day weighed food intake, 24-hour recall, and Fred Hutchinson Cancer Research Center/Block and Willett food frequency questionnaires) were tested for their ability to provide comparable food quotient and total energy expenditure data in doubly labeled water studies in 10 young and 10 older women. All methods gave mean values for total energy expenditure that were within 1% of each other. Individual values obtained using the 24-hour recall and food frequency questionnaires were within +/- 3% (standard deviation) of values determined using data from the 7-day weighed food record. These results suggest that it is not necessary to use time-consuming and expensive 7-day food records in doubly labeled water studies; instead, food intake data obtained more easily by 24-hour recall or food frequency questionnaire can provide comparable data. PMID- 9739804 TI - Rationale, process, and nutritional implications of peripheral blood stem cell transplantation. PMID- 9739803 TI - Comparison of dietary assessment measures in the Treatwell 5 A Day worksite study. PMID- 9739806 TI - A computerized method for determining outcomes in a food bank distribution system. PMID- 9739805 TI - Association of sociodemographic factors with barriers reported by patients receiving nutrition counseling as part of cardiac rehabilitation. PMID- 9739807 TI - Nutritional status of persons using a local emergency food system program in middle America. PMID- 9739808 TI - President's page: technology--from perception to commitment. PMID- 9739809 TI - Upskilling and dietetics professionals. PMID- 9739810 TI - Is Fanconi anemia caused by a defect in the processing of DNA damage? AB - Fanconi anemia (FA) is an autosomal genetic disease characterized by a complex array of developmental disorders, a high predisposition to bone marrow failure and to acute myelogenous leukemia. The chromosomal instability and the hypersensitivity to DNA cross-linking agents led to its classification with the DNA repair disorders. This review aimed at establishing whether it is still appropriate to consider 1/approximately FA within a DNA repair framework taking into account the recently discovered genetic heterogeneity characteristics of the defect (eight complementation groups). We discuss the possibility that the FA proteins interact to form a complex which may control different functions, including the processing of specific DNA lesions. Such a complex may act as a sensor to initiate protective systems as well as transcription of specific genes specifying, among others proteins, growth factors. Such steps may be organized as a linear cascade or more likely under the form of a web network. PMID- 9739811 TI - DNA single-strand breaks and DNA repair in the lymphocytes of wooden furniture workers. AB - DNA single-strand breaks (DNA SSB) in peripheral lymphocytes of wooden furniture workers and a control group, including smokers and nonsmokers, were detected by the microfiltration method. Our results show that cigarette smoking significantly increases the fragmentation of DNA single strands in the wooden furniture workers (by nearly two times) but not in the control group. Moreover, occupational exposure to wood dust and other wooden plant substances significantly induced DNA SSB only in the lymphocytes of smokers (by about two times). DNA repair in the lymphocytes was investigated as 3H incorporation into DNA. High 3H incorporation in the unstimulated lymphocytes of both smoking and nonsmoking workers, as compared to the references, suggests that besides DNA SSB other DNA damage can be caused by occupational exposure in the wooden plant. Since DNA repair is not always perfect, the possibility is high that the low level of DNA repair in the study group may lead to irreversible DNA damage. We think that the workers exposed to wood dust and the substances emitted by furniture coating materials in the plant may be at higher risk for mutagenesis and/or carcinogenesis than the unexposed population. PMID- 9739813 TI - Comet assay detects cold repair of UV-A damages in a human B-lymphoblast cell line. AB - During DNA repair studies, cells are occasionally kept on ice in order to suppress DNA repair. In the present studies cultivated human NC37 B-lymphoblasts were damaged by UV-A irradiation (365 nm) and DNA single strand breaks were detected at the single cell level with the alkaline comet assay in the temperature range from 4 degrees C to 44 degrees C. Single cell studies, in contrast to bulk experiments, allow to identify apoptotic or necrotic cells, which can be omitted for data analysis. Unexpectedly, similarly efficient single phase repair kinetics was found at all temperatures below 37 degrees C, i.e., particularly also in the cold. For recovery times below 20 min a linear decrease of DNA damage was detected. After 20 min, no additional repair was observed, i.e., complete repair of single strand breaks was not achieved. At 44 degrees C DNA damage increased with time, probably due to heat damage and cell death. Nucleotide excision repair inhibitors such as aphidicolin, 1-beta-D arabinofuranosyl cytosine (araC) and hydroxyurea, but not the base excision repair inhibitor methoxyamine caused a strong increase in DNA strand breaks. The use of repair inhibitors confirmed DNA repair at 4 degrees C. In conclusion, partial repair of UV-A damage is similar at 37 degrees C and 4 degrees C and is probably governed by nucleotide excision repair. Keeping samples on ice may not result in a total suppression of DNA repair. PMID- 9739812 TI - The CHO XRCC1 mutant, EM9, deficient in DNA ligase III activity, exhibits hypersensitivity to camptothecin independent of DNA replication. AB - We have analyzed the X-ray-sensitive CHO mutant cell line EM9 for sensitivity to the topoisomerase I inhibitor comptothecin. These cells exhibit defective repair of single strand DNA breaks. Recently, EM9 were complemented the DNA ligase III interactive protein, XRCC1. Defective XRCC1 apparently accounts for the low DNA ligase III activity that may explain the single-strand break repair deficiency of EM9 cells. Here, we demonstrate cytotoxic hypersensitivity of EM9 cells following a brief camptothecin treatment. Both the S-phase and non-S-phase populations of EM9 exhibited camptothecin sensitivity relative to the parent cell line AA8. In AA8 cells, only the 55% of the population corresponding to the S-phase subpopulation were sensitive to camptothecin, while the remainder of the population were totally resistant to doses as high as 10 microM. The role of DNA replication in the camptothecin sensitivity was studied using the DNA polymerase inhibitor aphidicolin in co-treatment with camptothecin. Aphidicolin treatment fully protected AA8 cells from camptothecin cytotoxicity. In EM9 cells, aphidicolin protected the S-phase fraction to some degree but all the cells remained sensitive to camptothecin cytotoxicity. These results suggest that EM9 cells are sensitized to camptothecin by a mechanism that is independent of DNA replication and may be a consequence of the XRCC1 mutation or the associated deficiency in DNA ligase III activity. Mechanistic models for the replication independent cytotoxicity of camptothecin in EM9 cells are discussed. PMID- 9739814 TI - 2-Hydroxyadenine, a mutagenic form of oxidative DNA damage, is not repaired by a glycosylase type mechanism in rat organs. AB - Oxygen radicals are known to play a role in causing cellular DNA damage, which is involved in carcinogenesis. 8-Hydroxyguanine (8-OH-Gua) is a major form of oxidative DNA damage and is known as a useful marker of DNA oxidation. Recently, we found another type of oxidative DNA damage, 2-hydroxyadenine (2-OH-Ade), which has a mutation frequency comparable to that of 8-OH-Gua. We compared the repair activities for two types of oxidative DNA damage, 8-OH-Gua and 2-OH-Ade, in 7 week-old male Sprague-Dawley (SD) rat organs. The repair activities were measured by an endonuclease nicking assay using 22 mer [32P]-end-labeled double-stranded DNA substrates, which contained either 8-OH-Gua (opposite C) or 2-OH-Ade (opposite T or C). In all of the SD rat organs we studied, the nicking activity for 2-OH-Ade was not detected, while that for 8-OH-Gua was clearly detected with the same conditions. Moreover, the 2-OH-Ade nicking activity was not induced in Wistar rat kidney extracts prepared after ferric nitrilotriacetate (Fe-NTA) treatment, which is known to increase 8-OH-Gua repair activity. These results suggest that 2-OH-Ade might not be repaired by the glycosylase type mechanism in mammalian cells. PMID- 9739815 TI - Mutational potentiality of stannous chloride: an important reducing agent in the Tc-99m-radiopharmaceuticals. AB - Stannous chloride (SnCl2) is frequently used in nuclear medicine as a reducing agent to label many radiopharmaceutical products with technetium-99m (99mTc). The aim of the present paper was to study the role of DNA repair genes in the repair of SnCl2-induced damage, using mutant strains of Escherichia coli lacking one or more DNA repair genes. Our results suggest that the product of the xthA gene, exonuclease III, is required for the repair of lesions induced by SnCl2. We further investigated the mutagenic properties of SnCl2 to a molecular level by using the supF tRNA gene as target in a forward mutational system. We have found that the survival of E. coli cells was strongly reduced with increasing concentrations of SnCl2. Moreover, when the shuttle vector pAC189 carrying the supF gene was treated with SnCl2, and then transfected to E. coli, we observed that its transformation efficiency dropped when compared to the non-treated control, with a parallel increase in mutation frequency after the damaged plasmids have replicated in bacterial cells. The mutation spectrum induced by SnCl2 reveals a high frequency of base substitutions, involving guanines. Sequence analysis of 41 independent supF mutant plasmids revealed that 39 mutants contained base substitutions, with 21 G:C to T:A and 17 G:C to C:G transversions. G to T transversions presumably resulted from 8-oxoG. However, the G to C one may be due to a yet unidentified lesion. PMID- 9739816 TI - Oxidative DNA damage induced by visible light in mammalian cells: extent, inhibition by antioxidants and genotoxic effects. AB - The extent of the indirect DNA damage generated in mammalian cells by visible light because of the presence of endogenous photosensitizers was studied by means of repair endonucleases. In immortalized human keratinocytes (HaCaT cells) exposed to low doses of natural sunlight, the yield of oxidative DNA base modifications sensitive to the repair endonuclease formamidopyrimidine-DNA glycosylase (Fpg protein) generated by this indirect mechanism was 10% of that of pyrimidine dimers (generated by direct DNA excitation). A similar yield of Fpg sensitive modifications, which include 8-hydroxyguanine, was observed in primary keratinocytes. The relative yield of oxidative base modifications decreased at higher light doses, probably as a result of photodecomposition of the endogenous chromophore involved. For the three cell lines tested, viz. HaCaT cells, L1210 mouse leukemia cells and AS52 Chinese hamster cells, the yield of oxidative base modifications generated by a low dose of visible light appeared to be correlated with the basal concentrations of porphyrins in the cells. Induction of cellular porphyrin synthesis by pretreatment with 5-aminolaevulinic acid increased the light-induced oxidative damage in L1210 cells several-fold. In both induced and uninduced cells, the damage was inhibited by more than 50% in the presence of ascorbic acid (100 microM), while alpha-tocopherol and the iron chelator alpha phenanthroline had no effect and beta-carotene even increased the damage. Even high doses of visible light did not significantly increase the numbers of micronuclei in L1210 cells or of gpt mutations in AS52 cells. The negative outcome can be fully explained by the photobleaching of the endogenous photosensitizers, which prevents the generation of sufficiently high levels of oxidative DNA damage. Therefore, the mutagenic risk arising from the indirectly generated oxidative DNA modifications induced by sunlight may be underestimated when results obtained at high doses are extrapolated to low doses or low dose rates. PMID- 9739817 TI - The use of oriC-dependent phage infection to characterize the ultra violet (UV) induced inhibition of initiation of DNA replication in Escherichia coli. AB - The oriC transducing phage lambda poriCc is a pseudovirulent phage capable of forming plaques on a lambda lysogen. This phenotype is dependent upon the presence of the oriC insert. The ability of lambda poriCc to form plaques on a lambda lysogen represents a potential phage assay system for studying aspects of oriC function. In the present study we establish that lambda poriCc infection of a lambda lysogen is a legitimate assay for oriC function. We use this assay to confirm the previously reported observation that initiation of DNA replication from oriC is transiently inhibited in a ultra violet (UV) irradiated cell at doses greater than 60 J/m2. We further demonstrate using this assay that the UV induced inhibition of initiation of DNA replication from oriC is not a SOS function nor a heat shock function. In the course of these studies, we found that lambda poriCc infection of a non-lysogenic cell is extremely sensitive to pre irradiation of the Escherichia coli host. We postulate that the sensitivity of lambda poriCc replication to host cell pre-irradiation reflects in some way the transient inhibition of initiation of DNA replication from oriC following UV irradiation. PMID- 9739818 TI - [The degrees of burns--classifications]. AB - Since 1996, a new three-degree classification has been adopted for determination of the depth of lesions caused by the thermal agent, published in Bulletin N(o) 7, XLII, 1996 of the Ministry of Health. The present report is aimed at facilitating surgeons in its practical implementation by making a comparative assessment of the various numerical classifications, employed both in this country and abroad, and adapting them to the three groups of nonnumerical classifications. In the field of scientific communication between doctors from different countries, the numerical classifications give rise to confusion and on account of that utilization of the verbal classification for evaluating the deepness of a burn lesion is strongly recommended. PMID- 9739819 TI - [Perforations of hepatic hydatid cysts]. AB - Out of a total of 753 patients operated for liver echinococcosis, in 43 (5.71 per cent) various perforations are observed, namely: in the biliary apparatus, chest, abdominal cavity, gastrointestinal tract, subphrenic and subcutaneous tissue. Their incidence in bile ducts is the highest. The lethality is rather elevated with two patients dying on the operating table. In conclusion, proceeding from the severity of the complication emphasis is laid on the necessity of early diagnosis of the disease, before development of serious complications, successfully achieved in the last few years. PMID- 9739821 TI - [Organ-saving, physiologically based and pathogenetically expedient operations on the stomach]. PMID- 9739820 TI - [Pancreatic cancer. The experience with its surgical treatment]. AB - Experience had with the surgical management of 483 patients, operated for cancer of the pancreatic gland in the Clinic of Abdominal Surgery over the period 1971 through 1994, is presented. Distribution of the patients by gender and age shows noteworthy consistency. Preoperatively, exact diagnosis in the clinic is made in 91.1 per cent, but usually the patients are admitted in advanced stage of the disease (91.6 per cent). Merely 42 cases (7.6 per cent) are free of metastases in regional lymph nodes or carcinomatous infiltration. The neoplasm is located in the head of the gland in 350 patients (66.5 per cent) with resectability attained in 10.1 per cent, in the body and tail--in 96 patients (19 per cent) with resectability 6.5 per cent; the neoplasm involves the total gland in 55 cases (10.2 per cent), and derives from the papilla in 25 (5.2 per cent) with operability 44 per cent. Operability for the total group amounts to 9 per cent, morbidity--17 per cent and overall lethality--15 per cent; of the latter following radical interventions 4 per cent, palliative interventions 8 per cent and explorative laparotomy 3 per cent. The details of the operative technique are comprehensively discussed. PMID- 9739822 TI - [The indications and choice of the technic for pancreatic resections]. AB - The progress of pancreatic surgery naturally leads to broadening the scope of indications for resection of the gland. Over the period 1970 through 1993, in the Clinic of Abdominal Surgery are performed 99 pancreatic resections for carcinoma of the pancreas (including cancer of papilla Vateri (45), endocrine-active tumor of pancreas (5), benign tumor (1), chronic pancreatitis (4), pseudocysts of the pancreas (4), cancer of an adjacent organ infiltrating the pancreas (36), benign lesions to a neighbouring organ (4). The size of resection depends on a number of factors of which location of the neoplasm and stage of the disease are the leading ones. Operations done: duodenopancreatic resection--47, left side hemipancreatectomy--13, resection of the body region and tail of pancreas--14, and partial resection--19. The choice of operative technique is discussed against the background of the variety of indications for pancreatic resection. Special attention is focused on the operative technique used in pancreatoduodenal resection. The practicability of performing pylorus-preserving intervention, the need of vagotomy and its scope are precisely determined. Also discussed are issues relating to the choice of organ for anastomosis with the pancreas and its protection, as well as the variants of successiveness of anastomoses with the biliary canal, residual pancreas and stomach. Morbidity and mortality rates show a decrease parallel to the surgical experience gained. PMID- 9739823 TI - [The surgical treatment of pancreatic pseudocysts following acute pancreatitis]. AB - The tactics and results of the operative treatment of pancreatic cysts, complicating severe destructive pancreatitis in a series of thirteen patients, are discussed. The following operative methods are made use of: marsupialization (1), Yurash (10), cystojejunoanastomosis with Braunova (2). The character and scope of surgical intervention are determined intraoperatively, depending on the anatomical situation faced. In pancreatic cysts operated according to Yurash (cystogastroanastomosis), an original drainage method with two probes introduced nasally is used--one wider into the anastomosis, and a narrower one into the duodenum for feeding. The probes are retained for periods ranging from 9 to 35 days. No relapse of the cysts operated by different methods are registered, with the exception of a female patient undergoing marsupialization. In one case operated according to Yurash where no preoperative preparation is done the outcome is fatal, with the patient dying of hemorrhage on the third postoperative day. All patients are operated within 3 months after the formation of cysts. The preoperative preparation includes Kontrikal, Petphtoruracil, atropine, heparin and antibiotic; in some patients the listed drugs are introduced intraarterially into truncus celiacus. A number of inferences are reached and recommendations made: 1. Waiting for the generally accepted 3-month term is unnecessary. 2. In cysts involving the head of the pancreas, tightly adherent to the posterior wall of the stomach, the method of Yurash with the modification suggested for probing should be given preference. 3. In cysts of the body region and tail cystojejunoanastomosis with Braunova is practicable. 4. Proceeding with the preoperative medication in the postoperative period is advisable. PMID- 9739824 TI - [Retroperitoneal laparoscopic adrenalectomy. The 3-trocar surgical technic]. AB - Laparoscopic adrenalectomy (LA), introduced in surgical practice quite recently, proves to be an effective method for treating diseases of the adrenal gland on account of the good visualization provided, in conjunction with all additional advantages of laparoscopic surgery, such as reduced traumatism, postoperative pain alleviation, shortened hospital stay and the like. Two patients presenting Cushing's syndrome are subjected to retroperitoneal laparoscopic adrenalectomy (RLA). With the patient in lumbotomy position on the operating table, a retroperitoneal space is created using a balloon dissector (Ethicon). The optics is introduced after placing a 10 mm balloon trocar (Origin). Under visual control two additional 10 mm trocars are placed along the course of lumbotomy line, accordingly along the anterior and posterior axillary lines. Next blunt dissection of the adrenal gland is done, and a clip applied to v. centralis. The adrenal gland is removed through one of the trocars, and the operation terminates after hemostasis and insertion of a drainage tube. The patients are discharged on the 4th postoperative day in a good general condition, mobilized, well fed with restored passage. In conclusion, the assumption is warranted that the early experience with RLA is encouraging as a safe and considerably less traumatic surgical procedure for operative management of diseases of the adrenal glands. PMID- 9739825 TI - [The indications for selective intraoperative cholangiography during laparoscopic cholecystectomy]. AB - The analysis covers 136 cases of laparoscopic cholecystectomy. Intraoperative cholangiography is performed in seven patients. Choledocholithiasis, missed during the operation, is diagnosed in one case only two months later. Preoperative ERCP is done in three patients, and postoperatively--in two. The follow-up study of the rest of patients, not subjected to intraoperative or endoscopic cholangiography, shows that up to one year after the operation there is not a single case of missed choledocholithiasis. The indications for intraoperative cholangiography are established on the ground of criteria worked out in the clinic. The obtained data confirm the hypothesis for selective application of the examination without augmenting the risk of missing pathological changes in the biliary ducts. PMID- 9739826 TI - [Genetic susceptibility to pancreatitis]. AB - Pancreatitis is a complicated polyetiological disease rather frequently met with. Inflammatory degenerative changes in the pancreas are the underlying cause of the condition which in acute cases may give rise to irreversible pancreonecrosis, and in chronic ones--to fibrosis development and severe pain syndrome. Of utmost importance is the patient's genetic susceptibility to pancreatitis. The purpose of the study is to assay the role of genetic factors involved in the etiopathogenesis of pancreatitis. The interest focused on alpha1 antitrypsin (alpha 1 AT) arises from the fact that its mutant forms are implicated in the destructive processes within the organism. The reduced inhibitory activity of alpha 1 AT enhances the action exerted by the proteolytic enzymes--trypsin and chymotrypsin [correction of hemotrypsin]. Impairment of the balance between proteases and their inhibitors plays certain role in pancreatitis development. Seventy patients, 44 men and 26 women, are covered by the study, with 42 of them presenting acute pancreatitis, and 28--chronic relapsing form. A high rate of alpha 1 AT mutant genes carrier state is established--14.28 per cent, exceeding statistically significantly the incidence of alpha 1 AT variants in the Bulgarian population--4.95 per cent (p < 0.01). In acute pancreatitis patients the incidence of alpha 1 AT variants is 2.38 per cent, and in chronic forms--32.14 per cent. In pancreatitis patients alpha 1 AT deficit brings about genetic predisposition to serious complications, e.g. chronification of the process. Individual therapeutic approach is mandatory, with Kontrikal used in chronic relapsing pancreatitis in the form of substitutive medication, and in acute pancreatitis--according to judgement depending on the clinical picture and laboratory findings. PMID- 9739827 TI - [Lithiasis of the extrahepatic bile ducts--its diagnosis and treatment]. AB - This is a survey paper assaying the relevance of clinical and laboratory data, and other conventional and currently used methods as well, and their precision in detecting concretions in the biliary apparatus, namely: endoscopic echography, magnetic resonance imaging, ERCP, PTH, CT etc. To increase the rate of concrement demonstration the diagnostic process should proceed intraoperatively by resorting to cholangiography, echography, choledochoscopy and the like. Nonoperative and operative methods of treatment are discussed, with the indications for nonoperative ones (endoscopic, extracorporeal lithotripsy, medicamentous and chemical litholysis) being listed, and their superiorities and side effects during practical implementation established. To improve the results of the latter eventually their combination is required. Various types of operative procedures are described, along with the indications for using them, advantages and shortcomings of each individual method, and practicability of their combined application under definite conditions. PMID- 9739828 TI - [A large postoperative hernia of the anterior abdominal wall with hernial contents--a sarcoma of the large intestine]. PMID- 9739829 TI - [Primary non-Hodgkin lymphoma of the anterior abdominal wall]. PMID- 9739830 TI - [A case of a symptomless amebic liver abscess manifested by respiratory failure]. PMID- 9739831 TI - [Dissection with an ultrasonic dissector during laparoscopic cholecystectomy]. PMID- 9739832 TI - [The comparative characteristics of methods for breast reconstruction after a mastectomy]. AB - Ablation of the breast because of cancer is a damaging operation leading the patients on a heavy psycho trauma. At the background of the main disease--cancer, deformation of the chest wall, caused by mastectomy their depression deteriorates. Therefore the breast reconstruction today is considered as an indivisible part of the complex treatment process. The techniques by means of witch the reconstruction could be achieved are: implantation of breast prosthesis, pedicle or free myocutaneous flaps. We enclose the comparative characteristic of the treatment results of 70 patients using different techniques. PMID- 9739833 TI - [Age-related indications for cheiloplasty in congenital and acquired defects of the upper lip--an anthropological study]. AB - Age-related changes in width (cheilion-cheilion) and height (subnasale-stomion) of the upper lip are studied in 2300 healthy Bulgarians, with ages ranging from three days to 102 years, for the needs of cheiloplasty in both surgery and fine arts. As shown by the results, at birth the upper lip in either gender appears to be one of the most developed organs of the human body, continuing to augment in size up to the eighth decade of life. In preschool age it terminates its intensive development, and in the puberty period it fails to exhibit the growth leap characteristic of the organism as a whole. Throughout the period of postnatal ontogeny, with the exception of early infancy, the lip in the male gender remains bigger, with the difference for height being rather markedly expressed during the ninth decade (3, 8 mm), and for width--thereafter (5.24 mm). The conclusion is reached that cheiloplasty may be undertaken regardless of the patient's age, during the early postnatal days inclusive, and the dimensions recorded for the age intervals being examined may serve as standard values. PMID- 9739834 TI - [Thymolipoma--a rare histological finding in patients with myasthenia gravis]. AB - Over a 15-year period, thirteen thymolipomas are histologically demonstrated in a total of 182 myasthenia gravis patients undergoing thymectomy. The incidence of this rare histological diagnosis in the aforementioned group appears to be considerably higher than the one so far reported in the pertinent literature. In the series reviewed men in advanced age prevail which is by no means typical of myasthenia patients. The postoperative results are very good, with not a single fatal outcome being recorded. There are basically three types of histological findings in thymolipomas--fat tissue alone, fat tissue with thymic tissue in involution, and fat tissue with thymic tissue. PMID- 9739835 TI - [The treatment results in 138 patients with breast prostheses]. PMID- 9739836 TI - [Odontogenic craniofaciocervical necrotizing fasciitis]. PMID- 9739837 TI - [Errors in the diagnosis of benign prostatic hyperplasia]. PMID- 9739838 TI - [Mathematical analysis in the prognostic determination of survivorship in patients with urothelial tumors of the upper urinary tract]. AB - Mathematical analysis of the probable five-year survivorship is performed in patients operated on for neoplasms involving the urothelium lining the upper urinary tracts. The prognosis groups thus obtained are compared with the results of 5- to 10-year-long observation on the postoperative survival in fourty-one patients. It is established that the anticipated survivorship is determined with a high-degree probability by resorting to the mathematical method described. PMID- 9739839 TI - [The surgical treatment of cervical lymph node metastases in differentiated thyroid carcinoma]. AB - Over the period 1980 through 1994, a total of 297 patients presenting histological evidence of differentiated carcinoma of the thyroid gland are studied. In thirty-two of them (10.8 per cent) palpable metastases in the lymph nodes are found at the time of making the diagnosis. A higher frequency of palpable lymph nodes is established in younger patients with histological diagnosis papillary carcinoma (62.5 per cent). In the 32 patients with palpable metastases in the lymph nodes, discovered during diagnosing of the disease, lymph node dissection is undertaken. The obtained results show that the modified cervical dissection yields favourable results, and at the same time preserves intact the neck structure. PMID- 9739841 TI - [Diagnostic laparoscopy in female sterility]. AB - Over a three-year period (1990-1993), a total of 410 diagnostic laparoscopies are performed. The series includes female patients with a minimum one year longstanding of sterility. Diagnostic laparoscopy is taken to be a routine method of making exact diagnosis in female sterility cases, and as a filtering procedure in forthcoming reconstructive operation or in the event of poor outlooks of eventual conception. The commonest finding is endometriosis (in 58.2 per cent), next ranking adhesions free of endometriosis (31.4 per cent) and bilateral obstruction of the uterine tubes (38.5 per cent). Endometriosis is usually located in the uterine body 27.5 per cent, uterine tubes 26.1 per cent and pelvic peritoneum 14.9 per cent. PMID- 9739840 TI - [Histological variants (subtypes) of differentiated thyroid carcinoma]. AB - This is a report on retrospective clinical and pathoanatomical study of seventy one patients presenting histological diagnosis differentiated carcinoma of the thyroid gland, undertaken with the purpose to identify the histological variants (subtypes) of differentiated thyroid carcinoma. As shown by the obtained results, the follicular variant is the commonest subtype of papillary carcinoma (41.18 per cent) followed by focal papillary carcinoma (25.49 per cent), and encapsulated variant (15.69 per cent). On the other hand, the most frequently met with follicular carcinoma subtype proves to be the least invasive variant (55 per cent). PMID- 9739842 TI - [The intraoperative determination of intestinal vitality with a fluorescent indicator]. AB - Intestinal obstruction due to strangulation is induced in dogs under experimental conditions, with intestinal wall vitality assessment done on the ground of standard clinical criteria, using fluorescence dye and UV rays, as well as histological study. Sensitivity, specificity and prognostic value of each of the methods employed are determined. The fluorescence method advantages are recorded, and the prospects of its clinical implementation are estimated. PMID- 9739843 TI - [Breast cancer and pregnancy]. AB - Breast carcinoma is a condition characterized by an ever increasing incidence. Young women too are ever more frequently involved. Currently a modern tendency is observed of young women to postpone conception for a later period of life on account of a number of socioeconomical reasons, which tendency tangibly augments the number of women affected with breast cancer during the pregnancy period. Most of the pregnant women are in the age group 34 to 35 years. Breast cancer during pregnancy is a malignant disease ranking second by incidence rate. The latter amounts to 10-30 per 100,000 of pregnant women. PMID- 9739844 TI - [Lithiasis of the extrahepatic bile ducts]. AB - This is a literature survey on issues relating to average incidence rate of choledocholithiasis for the last few years, basic mechanisms and factors underlying the formation of biliary calculi, as well as form and location of the concrements, with a special reference to the impact of the latter on the course run by the disease. Choledocholithiasis is divided in two forms--primary and secondary: in the former the underlying causes and criteria for the occurrence of calculi are described, and in the latter--the factors and conditions promoting migration of the concrements towards the bile ducts, with emphasis on the importance of the division suggested in choledocholithiasis treatment. The clinical picture with its characteristic symptoms, and the pathogenesis of the latter are also discussed. Evolution of the disease left without adequate intervention has unfavourable prognosis. A description is submitted of the various complications and their characteristic manifestations observed. Differential diagnosis is made both often and rarely met with diseases presenting similar symptomatology. PMID- 9739845 TI - [A postoperative urethral diverticulum in a man]. PMID- 9739846 TI - [Soft-tissue osteosarcomas: osteogenic sarcoma of the mesocolon causing ileus]. PMID- 9739847 TI - [A combination of hepatic echinococcosis and polycystosis]. PMID- 9739848 TI - [Postoperative anaerobic sepsis]. AB - As shown by clinical practice, postoperative anaerobic sepsis is a complication more common than usually thought of or microbiologically verified. The exceptionally difficult microbiological verification, regardless of the fact that original registered transport media are employed, is the underlying cause of obligate non-spore forming microorganisms from hemoculture being demonstrated in four patients only. In all of them Bacteroides fragilis is isolated and identified. Also, in all patients a fully developed clinical picture of sepsis is present along with the characteristic laboratory septic syndrome constellation as well. After pointing out the difficulties in diagnosing "post-operative anaerobic sepsis", and more particularly its verification, emphasis is laid on the clinical and laboratory symptoms presented by the patients, and on the important role played by the SIRS system, contributing greatly to an adequately oriented clinical thinking. PMID- 9739849 TI - [The lymphotropic marking and biopsy of sentinel lymph nodes in T1 tumors--a new approach to the staging of axillary lymphatic metastasis in breast cancer]. AB - It is the purpose of the study to assay the possibility of demonstrating axillary lymph status through marking and biopsy of sentinel lymph nodes. Preoperative lymphotropic marking of sentinel lymph nodes is performed in 48 female patients presenting mammary gland carcinoma, measuring up to two centimeters. Drimaren 0.5 ml (in 18 cases), Mitoxantrone (17 cases) or Patent Blue (13 cases) each are perineoplastically injected in two points. Intraoperatively, in 34 patients blue stained lymph nodes (from Patent Blue and Mitoxantrone mainly) are identified, and compared with the results of axillary dissection. Metastases in sentinel nodes are documented in seven instances. In the remainder (30) which are histologically negative, lymph node metastasis II level is discovered in one case (false negative = 3.3 per cent). As shown by the initial observations perioperative marking of lymph nodes with Patent Blue and Mitoxantrone contribute to demonstrate sentinel nodes, and by biopsy study of the latter it is possible to judge about the pattern of lymph metastasizing of mammary gland carcinoma. PMID- 9739850 TI - [A paraneoplastic syndrome in patients with a carcinoma of the upper urinary tract]. AB - Evidence of a paraneoplastic syndrome is established in 21/63 patients (33.3 per cent) presenting upper urinary tract tumors. It becomes manifest 2-5 months before diagnosing the basic malignant disease. Subfebrility and anemia are the commonest paraneoplastic symptoms, clinically expressed in 15.87 per cent of cases. As the result of radical treatment, this symptomatology disappears within 10 days to two months after resorting to radical surgery. The paraneoplastic syndrome may be interpreted as a clinical "tumor marker;, indicating whether or not the treatment is radical, or the occurrence of a relapse and metastases may be anticipated. PMID- 9739851 TI - [Liver resections and operations in portal hypertension viewed via the experience of our practice]. AB - After listing the indications for liver resection, the operative technique used is discussed with a special reference to an original modification implemented in practice. The case material is made of 52 liver resections and 6 lobectomies with a favourable outcome. In one lobectomy with subtotal proximal resection jejunogastroplasty is performed, supplemented by isolated antireflux anisoperistaltically interposed invagination esophagojejunostomy, duplicated by suturing its two portions. Application of catheter, inserted into the recanalized umbilical vein, is described under the heading of locoregional chemotherapy. In carcinomas involving the confluence of hepatic ducts a variant of Rodney Smith's operation is used while in hepatocholedochus resection plastic repair is done over Kerr drainage. In portal hypertension, after discussing the various methods existing, attention is called to the indirect shunts: splenectomy with omentoreno and omentoparietopexy, Charsky and fenestration of Glisson's capsule using electric knife and argon with omentohepato- and hepatoparietopexy, as well as implantation of the recanalized v. umbilicalis into m. rectus abdominis dexter. In 15 cases operated on by parenchyma stimulating procedures, survivorship ranging from 3 to 26 years is achieved. For the purpose of prophylaxis against cholangiohepatitis and cirrhosis the following operations are performed: 68 papillotomies and plastic repairs over balloon catheter, passed through the choledochus and papilla, using an electric knife, 12 choledochoduodenostomies type "frog mouth", and 11 deperitonizations-desympathizations with a successful cure being attained. PMID- 9739852 TI - [Splenectomies for Hodgkin's disease--a study of the period 1984-1994]. AB - Explorative laparotomy with ensuing splenectomy in Hodgkin's disease is applied for the first time in the Stanford University in the late 60-ies. Opinions on the pros and cons of the method in terms of number of cases, complications and the like are largely dissenting. The analysis covers 153 patients with Hodgkin's disease subjected to splenectomy. Primary diagnosis is made on the ground of lymph node biopsy. Wide spreading of the disease, coincidence of diagnoses in different locations and complications are briefly discussed, with recommendations made concerning improvement of the organization of sample handling and correlation with the clinical findings. PMID- 9739853 TI - [Reoperations on the biliary system]. AB - Reoperations of the biliary apparatus is a branch of biliary surgery still not well enough clarified and difficult to cope with. A total of 169 patients are subjected to operation and investigation in the clinic of abdominal surgery. In 71 of them reoperation is undertaken over the period 1952-1973, and in 97--in the period 1974 through 1993, representing 8.7 per cent of all biliary operations done for benign diseases of the biliary apparatus. In 95 per cent of cases the primary operation is related to cholelithiasis (ChL). One-hundred fifty-five cases (92.6 per cent) are reoperated once, nine (5.3 per cent)--twice, and four (0.7 per cent)--three, four and five times. What is more, 56 of the patients are operated in the clinic of abdominal surgery, and 112--elsewhere in surgical units and departments throughout the country. The severer clinical picture, prolonged postoperative period, increased operative risk and worsened prognosis in the latter group are underscored. The underlying causes necessitating secondary corrective intervention are analyzed--76 per cent are conditioned by ChL, and 24 per cent--by the primary operation. The indications for reoperation are classified in three groups: a) failure to remove or partially removed gallbladder, b) in case of preexisting primary, or secondary postoperative development of various forms of cholelithiasis, c) in surgery induced morbid conditions. A table is presented illustrating the character of secondary operations, performed in the series of 168 patients under study, namely: in 20 per cent the gallbladder is operated on, and in 80 per cent--the extrahepatic bile ducts. Postoperative morbidity is higher as compared to the one in primary operations and not infrequently it is conditioned by preexisting complications. Postoperative mortality rate amounts to 10 per cent. PMID- 9739854 TI - [Planning the TRAM flap in reconstructive breast surgery after mastectomy. Transverse rectus abdominis muscle]. AB - The guidelines for postmastectomy breast reconstruction are the contralateral healthy breast. We tend to provide symmetrical reconstruction without or with minimal modification of the contralateral breast. The technique we are using for preoperative planning allows immediate de-epithelialization and tailoring of the dermoadipose extensions of the flap at the beginning and not at the end of the operation. This reduced operative time and the need for secondary correction and contralateral mammaplasty. PMID- 9739855 TI - [Organization, diagnosis and the treatment procedure in chest, abdominal and combined trauma]. AB - Thoracic and associated injuries pose problems demanding enormous interdisciplinary efforts regardless of the improved organizational, diagnostic and treatment potentialities. As shown by the experience of the Emergency Surgery Section at the Pirogov Institute, rendering medical services to patients presenting chest and associated injuries require coordinated integration of specialists from various profiles along with specification of the priorities and hierarchy of the emergency measures undertaken. Over a 12-year period (1985 through 1996), a total of 6983 patients with chest, abdominal and multiple trauma are admitted. They are distributed as follows: chest trauma casualties--3286, abdominal trauma--679, and multiply injured--3018. Chest injuries are divided in close and open injuries--2843 and 444, respectively; the latter include 29 gunshot and 415 penetrating-incise wounds. The abdominal trauma group includes 679 cases, distributed as follows: spleen rupture--341, disruption of liver and mesenterium--151, and lesion to a hollow abdominal organ--187. PMID- 9739856 TI - [The correlation between serum thyroglobulin and iodine-131 scanning in detecting metastases in patients with a differentiated thyroid carcinoma]. AB - One-hundred seventy-one patients, operated for differentiated thyroid gland carcinoma, are subjected to serum thyroglobulin assessment and whole-body scanning. In twenty-nine of the total of 171 patients there is evidence of relapses and metastases, discovered by either examination. In 27/29 cases with metastases (positive scan finding) a high serum thyroglobulin level is documented. The correlation between serum thyroglobulin level and whole-body scanning finding among patients without residual normal thyroid parenchyma (status post total thyroidectomy) is much more favourable than the one in patients presenting normal residual parenchyma (status post organ-salvaging operation). The conclusion is drawn that serum thyroglobulin contributes greatly to the diagnosis of recurrences and metastases, especially in patients with normal residual parenchyma missing. PMID- 9739857 TI - [Anorectal manometry--the first impressions of its use in pediatric surgical practice in the diagnosis and treatment of chronic obstipation]. AB - The first impressions from anorectal manometry application in the diagnosis, differential diagnosis and treatment of chronic constipation in children is discussed. In the pediatric surgeon's practice the method may be used as a screening examination, contributing to identify the underlying organic causes of the constipation syndrome. What is more, manometric examination has an essential practical bearing on the clinical therapeutic approach to various forms of habitual chronic constipation. PMID- 9739858 TI - [Primary lymphoma of the stomach]. AB - Primary lymphoma of the stomach (PLS) is an uncommon neoplasm, accounting for up to 8 per cent of all malignant neoformations with this particular localization. Its development is linked to the so-called mucosa associated lymphoid tissue (MALT), wherefrom its denomination--MALT lymphoma--is derived. In the stomach, under ordinary conditions, it is nonexistent, and in most of the cases it is acquired through infection with H. pylori. In a prevailing percentage of PLS it is a matter of B-cell lymphoma with low-degree malignancy. T-cell lymphomas occur sporadically, as well as primary Hodgkin's lymphoma which is exceptionally rare. Histologically the B-cell lymphoma reiterates the structural pattern of Peyer's plaque prototype, involving also a number of non-neoplastic components. The most distinguishing cells are reminiscent of centrocytes (centrocyte-like cells--CCL)- a term adopted for all MALT-lymphoma cases. They vary considerably in terms of cytological appearance, and may even bear resemblance to Hodgkin's and Sternberg's cells. Signet-ring lymphomas, known in several variants, are also by no means ruled out. Immunohistochemical typing presupposes a definitive diagnosis being made. In case of immunosuppression (transplantation, HIV-infection) the risk of B-cell lymphomas development augment, and what is more, most of them associated with Epstein-Barr viruses characterized by a substantially more aggressive course and poor prognosis. PMID- 9739859 TI - [Primary malignant appendiceal tumors]. AB - Primary appendicular malignant tumors are rarely met with, and account for 0.5 per cent of all intestinal tumors. Of them 99 per cent are assigned to four types of neoplasms--carcinoid tumors, mucinous cystadenocarcinoma, adenocarcinoma and adenocarcinoid. These tumors are discussed under a separate heading owing to their location, clinical manifestation and characteristic features of the therapeutic approach. All patients presenting malignant neoplasms should be mandatorily subjected to follow-up study, insofar as 15-20 per cent of them develop secondary malignant processes located in the gastrointestinal tract mainly. Appendectomy is a sufficient in size operation done in carcinoids up to 1.0 cm in diameter; right hemicolectomy is indicated in carcinoids exceeding 2 cm in diameter; in adenocarcinoma measuring 1 to 2.0 cm, the extensiveness of intervention depends on the presence of infiltration into the mesoappendix. In mucinous cystadenocarcinoma right hemicolectomy is the operation of choice. In adenocarcinoma and adenocarcinoid tumor right hemicolectomy is proposed as a radical intervention, in conjunction with oophorectomy in menopausal patients. PMID- 9739860 TI - [A giant apudoma of the pancreas]. PMID- 9739861 TI - [A case of pancreatic polycystosis]. PMID- 9739862 TI - [The indications for pancreatic resection for a case of microcystic adenoma of the pancreas]. PMID- 9739863 TI - [Osteosarcoma of the coccyx]. PMID- 9739864 TI - [Chest injuries treated in the town hospital of Sabha, Libya]. AB - This is a report on 113 patients with chest injuries subjected to treatment over the period 1991 through 1993. Of them 72.5 per cent are with ages ranging from 11 to 30 years, and 94.7 per cent are men. The series includes cases presenting incised penetrating wounds (49.6 per cent), involvement in motor-vehicle road accidents (26.5 per cent) and gunshot wounds (15.9 per cent). Of the total number 23 per cent are polytraumatized. Most of the patients (56.7 per cent) have hemopneumothorax with pleural drainage being the only treatment applied. In 15.9 per cent thoracotomy, laparotomy or thoracotomy plus laparotomy are used. Complications are recorded in 7.1 per cent, with deaths amounting to 2.7 per cent. PMID- 9739865 TI - [Rib fractures and flail chest in closed chest trauma--stabilizing operations and the results]. AB - A contingent of patients presenting closed chest injuries, sustained over a 12 year period (1985-1996), are analyzed. Of the total of 6938 traumas, chest injuries amount to 3286 (47.06 per cent) of which 2842 (40.7 per cent)-closed. Of the latter 2248 (79.09 per cent) are located in the left thoracic half, 420 (14.77 per cent)-in the right thoracic half, and in 174 (6.12 per cent) it is a matter of bilateral involvement. At the point of heaviest impact, involving 5th to 9th rib segment, left side injury is recorded in 2034 cases (71.56 per cent), right side--408 (14.35 per cent), and bilateral--in 155 (5.45 per cent). There are 28 fractures of the sternum (0.85 per cent). Of the latter 14 are located in the manubrium sterni region, twelve--in the middle portion, and two in the distal part of the sternum. Over the last 5 years (1991 through 1995), of 63 casualties with flail chest 16 are with indications for stabilization osteosynthesis, and accordingly subjected to operation. A Schimelmann plate is used in 13 cases, artificial rib type "Ampar"--in three, and sternal stabilization--in one. PMID- 9739867 TI - [The postoperative results of the surgical treatment of hepatic echinococcosis using a closed surgical method]. AB - The authors examined critically no big one series of 41 patients undergone surgical procedures about liver hydatid cyst after so called "covered method". The simplicity of the method is signified and the lower risk, mortality and morbidity makes it convenient for surgeons with different degree and ability. The post operative application of 30% NaCl solution remove the risk of protoscolices into the fibrous capsule. PMID- 9739866 TI - [Central neurocytoma--the need for reassessment of histological diagnoses in tumors in the lateral and third ventricles]. AB - J. Hassoun and co-workers (1982) described two cases presenting intraventricular tumors of nervous origin, coined by them with the term "central neurocytoma" of the ground of electron microscopic data. As shown by the studies of other authors, central neurocytoma represents a separate morphological entity having light- and electron-microscopic and immunohistochemical characteristic of its own. In the recently published WHO classification of brain tumors (1993) central neurocytoma is entered as a tumor presenting new morphological patterns. Central neurocytoma occurs mainly in young persons, develops in the lateral and third ventricle, comparatively well differentiated from surrounding structures, rich in calcifications, disclosing proper relative specificity during CT scanning and MRI study. Light-microscopically this tumor bears resemblance to, and is erroneously diagnosed as oligodendroglioma or ependymoma. Proceeding from retrospective reassessment of two patients with intraventricular tumors and an additional observation, and on the ground of pertinent literature data, the morphological, clinical and nerve imaging characterization of central neurocytoma is outlined--a tumor met with in the daily routine practice, but usually erroneously interpreted. PMID- 9739868 TI - [Emergency reoperations in surgery of the gastrointestinal tract]. AB - Data concerning the therapeutic approach in a series of 28 patients, representing 1.3 per cent of the total number of abdominal operations performed in the Chair of Propedeutics of Surgical Diseases over the period 1990-1995, are retrospectively analyzed. In 19 of them primary operative interventions are done on an emergency basis. In further 19 cases the complications have inflammatory character. A total of 31 operations are done with lethality amounting to 36 per cent (10 patients). Multiorganic insufficiency develops in five of them (mean age 39 years). The remainder, dying of septic complications, are with mean age 76 years. The successful treatment of intraabdominal postoperative complications is largely dependent on the surgeon's experience and skills, material equipment and facilities available and on the algorithm of the therapeutic approach adopted. PMID- 9739869 TI - [The diagnostic and clinical aspects of postoperative dysphonias following operations on the thyroid]. AB - Postoperative dysphonia is not rare and in some cases life-treating complication in the thyroid surgery. Its emerge, autonomous, or attended with dysphagia and dyspnea is an early symptom for intraoperative lesion of n. laryngeus recurrens. The investigation is carried out over 21 patients with postoperative dysphonias, operated for euthyroid or hyperthyroid strumas and followed-up for a period of 3 10 months. Analysis of the results indicated that the main reason for postoperative dysphonias in thyroid surgery is the lesion of n. laryngeus recurrens. Along with this one is impressed by the fact that in 30% of the cases dysphonic complaints may owe to nonsurgical reasons. PMID- 9739870 TI - [Bronchoscopic assessment algorithms for the practical evaluation of the rheological properties of the tracheobronchial secretion and the classification of the degree of the disordered drainage function of the tracheobronchial tree (TBT) in chest and combined trauma with chest trauma as the leading injury]. AB - Ventilation impairment, due to ineffective elimination of the mucous-hemorrhagic content from the tracheobronchial tree (TBT), obstructs the upper airways with the ensuing ventilation reduction giving rise to atelectases and progressive alveolar block. There is evidence of transudation and exudation into the pulmonary pathways and pleural cavity. A series of 276 patients presenting closed chest trauma are subjected to fibrobronchoscopy (FBS) and follow-up study. In 92 of them bronchoscopy is performed 2 to 15 times per patient, accordingly: in 75 twice, in 10-five times and in 15-twice. One-hundred twenty-nine of the total of 276 cases under study are on mechanical ventilation. In 56 instances FBS is carried out through a tracheostomy cannula, in 73-by intubation, in 18-through the mouth, and in two--through the nose. Based on the obtained results, algorithms for assessment of the rheological properties of tracheobronchial secretion and degree of impairment of TBT drainage function during emergency FBS in closed chest injuries are worked out, having an essential practical bearing on the diagnostic and therapeutic approach to closed thoracic trauma. PMID- 9739871 TI - [Volvulus of the small intestine]. AB - Volvulus of the small intestine is a condition of bowel obstruction due to knotting and twisting of the small intestine. Two types of volvulus are described: 1) primary small intestinal volvulus where no predisposing factors exist, and 2) secondary volvulus where congenital or acquired conditions promote twisting of the small intestine. Over a 5-year period, 18 patients (eleven men and seven women) presenting volvulus of the small intestine are operated in the Emergency Surgery Clinic of the University Hospital "Queen Giovanna", representing 8.7 per cent of the total of 206 cases of small intestinal mechanical ileus (incarcerated herniations involving the small intestine are not included in the series). Primary volvulus is found in one patient. In those presenting secondary volvulus adhesions are the commonest underlying cause of small intestinal rotation--13 cases, next ranking primary tumor of the small intestine--one case, Meckel's diverticulum--one, carcinosis of peritoneum--one, and one patient with small intestine volvulation around colostomy. The most frequently encountered symptoms and laboratory examinations performed are analyzed. Intestinal necrosis is established in four instances (22 per cent). One patient dies of peritonitis and polyorganic insufficiency. Volvulus of the small intestine should be mandatorily considered in patients presenting mechanical ileus of the small intestine. Early operative intervention is a therapeutic approach contributing to preclude intestinal necrosis. PMID- 9739872 TI - [Practical management guidelines in closed craniocerebral trauma]. PMID- 9739873 TI - [The use of radiopharmaceuticals for myocardial perfusion in oncological diagnosis]. AB - Recently, radiopharmaceutics for myocardial perfusion are widely used in oncological diagnosis, not merely supplementing the other methods of examination, but having also an independent role both in terms of making exact diagnosis, and in evaluating the diffusion of malignant tumors, as well as in estimating the results of the therapy undertaken. Their widespread implementation in practice is still not well enough studied, and awaits further clarification. PMID- 9739874 TI - [Hemangiopericytoma of the omentum--a review of the literature and a case report]. PMID- 9739875 TI - [A case of a synchronous tumor of the large intestine combined with a leiomyoblastoma of the small intestine with a course of subileus]. PMID- 9739876 TI - [Strangulated small intestine ileus due to a leiomyoma (a case report)]. PMID- 9739877 TI - [A case of ischemic colitis in a young woman developing against a background of vasculitis of unknown etiology]. PMID- 9739878 TI - [Bilithiasis (cholenephrolithiasis)]. AB - The study covers 232 patients with chronic calculous cholecystitis, operated in the Surgical Clinic of the Military Hospital--Plovdiv over the period 1991 through August 1995. Bilithiasis (cholenephrolithiasis) is diagnosed in 26 cases (11.2 per cent). This is a condition running a clinical course characterized by pain in the right subcostal and lumbosacral regions, accompanied by nausea and vomit in 57.7 per cent. Eighteen patient sustain renal crisis. Palpatorily, pain is established in the right subcostal region, and positive succussio renalis--in twelve patients. In nine patients there is evidence of albuminuria and leukocyturia (34.6 per cent). In 26 cases the echographic study reveals presence of concretions in the gallbladder, and in all of them-renal calculi too (left kidney--9, right kidney--9, and bilaterally--8 patients). Intraoperatively, concretions in the gallbladder are found in all patients, with hydrops and empyema of the gallbladder documented in five, and vesicoduodenal fistula in one (23 per cent), whereas in the patients free of urolithiasis they amount to 10.8 per cent which points to a reciprocal aggravation of the two pathological conditions. The early, as well as the long-term results of the operative management applied are estimated as very good. PMID- 9739879 TI - [Germ-cell tumors of the brain--their course and management]. AB - Germ-cell tumors (GCT) of the CNS constitute a small group with an incidence varying in the range 0.1 to 3.4 per cent of all primary brain neoplasms. In the WHO classification of brain tumors they are differentiated in a separate group, and further subdivided into: germinoma, teratoma (mature, immature, malignant), embryonal carcinoma, choriocarcinoma, endodermal sinus tumor and mixed germinal tumors. Of them germinoma is the most frequently encountered (41-65 per cent of all GCT), whereas the incidence rate of mixed GCT may reach 32 per cent Mainly median structures are being involved--III ventricle (pineal region and hypothalamo-hypophyseal structures), basal ganglia, with isolated cases being described in corpus callosum, cerebellum, medulla oblongata and spinal cord. Eleven patients with a total of 13 verified GCT are presented (in one instance germinomas involving the pineal body and suprasellar region are simultaneously established). The clinical course data, results of the diagnostic examinations done (CT, brain angiography, tumor markers), treatment carried out--surgical, radio- and chemotherapy, as well as the outcome of the long-term follow-up study of patients are analyzed. Three patients have mixed GCT. It is of interest to note a case where ten years after the removal of a benign teratoma in the pineal region, germinoma in the same location is diagnosed and removed. PMID- 9739880 TI - [Spontaneous internal biliary fistulae--a report of 4 clinical cases]. AB - This is a report on personal experience had with operative treatment of chronic calculous cholecystitis. Of the total of 228 patients operated on, spontaneous internal biliary fistulas are discovered in 4 cases (1.75 per cent): Mirizzi's syndrome--1 case. Fistula cholecystoduodenalis--3 cases. The development of fistulas is attributed to the elevated intrabladder pressure and activation of the infectious process and destructive changes taking place in the gallbladder wall, with ensuing breakdown and possibility of the concrement to penetrate an adjacent organ. It is presumed that pain localized in the right subcostal area and epigastrium is the symptom most strongly expressed. Cholecystocolonic fistula runs the heaviest clinical course among the patients operated on. PMID- 9739881 TI - [The treatment problems of soft-tissue infections]. AB - The treatment results is a series of twenty-six patients running a very serious course of soft-tissue infections are analyzed. Of the total number ten have diabetes mellitus. The sepsis severity score is 23 average according to the APACHE II rating system. Operative management is undertaken without any delay. It is preceded by comprehensive correction of the metabolic disorders present and antibiotic treatment, initially substantiated by presumable microbial causing agents. A total of 62 operations are done. In 36 per cent of cases being examined, the microbiological study contributes to a proper orientation of the antibiotic therapy performed. Bacteremia is documented in three instances. Total mortality rate amounts to 32 per cent (8 patients). Infections running a serious clinical course may be caused by one or more microorganisms, but in either case the lethality is noteworthy. Active surgical management may prove effective provided it is initiated in the most opportune time. It is only part of the complex therapeutic approach to sepsis. PMID- 9739882 TI - [Problems in the surgical treatment of diabetic gangrene]. AB - Parallel to life expectancy prolongation and constantly improving life style of diabetic patients in this country, observed during the last decade, the problems relating to complications in diabetes become of primary importance. From 60 to 70 per cent of the diabetics with foot ulcerations have neuropathy. The serious forms of nerve involvement appear to be a major cause of lower extremity amputation. Analysis of the clinical case material, covering 382 patients with diagnosis diabetic gangrene, is done with the purpose to reduce the percentage of disabling high-level amputations, and determine with precision the scope of operative activity, indications for operative intervention and type of surgical procedure in diabetic patients. A total of 58 major amputations are performed, with their overall rate amounting to 13.6 per cent. The main trends of the therapeutic approach to diabetic foot are aimed at performing sparing, economically more beneficial operations, not leading to invalidization. PMID- 9739883 TI - [Diaphragmatic injuries in thoracoabdominal trauma]. AB - In closed thoracoabdominal trauma the diagnosis rupture of the diaphragm is usually made in 5 per cent of the casualties. Over a 12-year period (1985 through 1996), in the Pirogov Institute are admitted 3018 cases presenting polytrauma. Among the contingent of closed thoracoabdominal trauma lesions to the diaphragm are registered in 151 cases, and among those with open injuries--in 21 cases. There are 71 closed injuries in PTP, and 80 closed thoracoabdominal injuries caused by high falls. Open injuries associated with lesion to the diaphragm are due to gunshot wounds in two cases, and inflicted by knife and other pointed objects in nineteen. In closed trauma there is 2:1 male-to-female ratio, and in open injuries--5:1. PMID- 9739884 TI - [Practical guidelines for the management of closed spinal cord injuries]. AB - The principles underlying the necessity of operative management in handling closed spinal cord trauma, its indications and types of operative interventions are comprehensively discussed. Under separate headings are described the peculiarities of the approach to injuries involving cervical, thoracic and lumbal segments of the vertebral column, with a summarization of currently used methods and procedures, contributing to the more complete functional recovery of the spine, preservation and creation of optimal conditions for regaining the functions of the spinal cord affected which in turn, renders easier the performing of daily living activities, precludes the development of a variety of complications in the body as a whole, and first and foremost, allows for early initiation of rehabilitation measures. PMID- 9739885 TI - [Hepatic echinococcosis--the complicated forms. The clinical picture, diagnosis and treatment]. AB - This is a survey paper on the current trends in diagnosing and operative management of the complicated forms of echinococcus of the liver. The possibilities offered by the puncture-aspiration method, applied under echographic and computerized-tomographic control are assayed. Also discussed is the role and place of chemotherapy against the background of combined therapeutic approach. PMID- 9739886 TI - [The molecular biology and genetics of colorectal carcinoma]. AB - There is vast evidence in support of the idea that accumulated genetic changes (mutations) are the underlying cause of neoplasia development. This multi-step process is aptly illustrated by colorectal carcinoma (CRC), usually developing in the course of decades, and presumably requiring at least seven genetic events to complete its development. In CRC the oncogenes most frequently undergoing mutation are c-k-ras and c-myc, and among tumor suppressant genes--APC, MCC, DCC, p53. An updated model of the molecular bases for adenoma occurrence and its evolution into carcinoma is presented. Inheritance of a single gene only which has undergone mutation augments substantially the predisposition to CRC. This is noted in a clearcut manner in the hereditary syndromes familial adenomatous polyposis (FAP) and hereditary non-polypous colorectal carcinoma (HNPCC). Recent studies along these lines suggest that the genetic defect in FAP increases the incidence of tumor initiation through functional impairment of the APC gene which is a gene regulator of the enhanced colorectal mucosa proliferation. Contrarily, the defect in HNPCC involves mainly the tumor progression through mutation of the DNA repair genes (MMRs), which are regulators of the genome stability. The study of hereditary syndromes give rise to a new concept for the occurrence and development of sporadic and inherited cancer in humans. PMID- 9739887 TI - [New aspects in the etiopathogenesis of colorectal cancer]. PMID- 9739888 TI - [Restitution in patients with colorectal carcinoma after the Hartmann operation]. PMID- 9739889 TI - [The treatment of postoperative large-intestine peritonitis with a laparostomy kit]. PMID- 9739890 TI - [Current antibiotic prophylaxis in colorectal surgery]. AB - Over a one-year period (November 1996 to November 1997), in the Emergency Surgery Clinic perioperative parenteral antibiotic prophylaxis during colorectal operations is performed in a series of 32 patients, admitted on an emergency and deferred urgency basis, or for routine operative treatment. Of them 30 present malignant processes involving colon and rectum, and two--inflammatory diseases. All patients receive single i.v. injections with Cephalothin/Cefazolin at dose 2.0 g and Metronidazole 0.5 g immediately after anesthesia induction. In nine cases additional early treatment is necessitated--within 24 hours after the operative intervention--by administration of the same drug combination 4 times at 6-hour intervals, and in another two prophylaxis is substituted for continuous 5 day treatment using the same therapeutic scheme. In 30 patients the postoperative period runs a course free of noteworthy complications. In two instances there is evidence of operative wound suppuration, and in further two--urinary tract infection development unrelated to the antibiotic prophylaxis applied. The specific features characterizing the application of antibiotic prophylaxis during colorectal surgery are discussed, and appropriate drug therapy schemes are recommended, consistent with worldwide and Bulgarian experience along this line, as well as with the concrete hospital and economical conditions in this country. Special emphasis is laid on two aspects: optimization of the timing of antibiotic agent/agents injection, and reaching peak serum and tissue bactericidal concentrations in the immediate vicinity of the surgical incision; optimization of the duration of antibiotic prophylaxis on the ground of well established indications for the application of antibiotic prophylaxis in colorectal surgery in compliance with the dynamic patterns of intra- and postoperative septic risk. The modest number of patients subjected to updated parenteral perioperative antibiotic prophylaxis does not warrant a definitive interpretation of the data from the comparative clinical and pharmaco-economical analyses performed. Nevertheless, the preliminary results point to the economical expedience of the therapeutic approach suggested. PMID- 9739891 TI - [Postoperative peritonitis]. PMID- 9739892 TI - [Intraoperative lavage and primary anastomosis under emergency conditions in obstruction of the large intestine]. PMID- 9739894 TI - [Known and potential risks related to the use of high dose intravenous immunoglobulins]. PMID- 9739893 TI - [The indications and technic for Whitehead's operation in hemorrhoidectomy]. PMID- 9739895 TI - [Pristinamycin versus oxacillin in the treatment of superficial pyoderma. A multicenter randomized study in 293 outpatients]. AB - BACKGROUND: Superficial pyoderma occurs frequently. Generally, the benign infection is caused by Staphylococcus aureus and/or a group A streptococci. The subject is controversial, but treatment usually is based on narrow-spectrum antibiotics active against both germs. PATIENTS AND METHODS: A multicentric, randomized, double-blind, double-placebo study was conducted to compare pristinamycin (1 g b.i.d.) with a reference antibiotic, oxacillin (1 g b.i.d.) for 10 days. Inclusion criteria were: both sexes, age 15-80 years, clinical diagnosis of superficial pyoderma (impetigo, wound infection within the last 15 days, furunculosis, carbuncle, perionyxis), informed consent. The general practitioner investigators (n = 52) were assisted by 9 dermatologist coordinators. Clinical diagnosis was validated by a committee of experts at the end of the study after analyzes of the photos and bacteriological results obtained on samples taken at the practitioner's office on visit 1 (D0), visit 3 (D14 +/- 3) and visit 4 (D25 +/- 3). Successful treatment was defined by clinical, bacteriological and photographic efficacy at visit 3 (equivalence analysis: one-way 95 p. 100 confidence interval). RESULTS: There were 293 included patients given pristinamycin (n = 151) or oxacillin (n = 142). Mean age of analyzed patients was 40 +/- 17 years. Diagnosis was confirmed in 255 patients in accordance with the protocol: furunculosis or carbuncle (n = 100), recently superinfected wound (n = 97), impetigo (n = 41), acute perionyxis (n = 17). Thirty-five patients (12 p. 100) were considered to have been wrongly included. The germs most often isolated were: Staphylococcus aureus (n = 126), group A streptococci (n = 13), group B streptococci (n = 5) and P. multocida (n = 3). At visit 3, the two treatments were found to be equivalent with a success rate of 86.7 p. 100 for pristinamycin and 89.8 p. 100 for oxacillin (CI [*9.97]). Tolerance was statistically comparable between the two treatments (27 to 32 percent minor side effects). DISCUSSION: This study is the first performed in outpatients attended by general practitioners with diagnostic confirmation on both bacteriological and photographic evidence of superficial pyoderma. The results obtained demonstrate the good reliability of such studies although 12 p. 100 of the patients were wrongly included, a factor which should be taken into account for future studies. The efficacy and tolerance of pristinamycin were statistically equivalent to those of oxacillin for all the patients with superficial pyoderma. Nevertheless, the subgroup of patients with folliculitis gave rather heterogeneous bacteriology and therapeutic results. PMID- 9739896 TI - [Self-induced cutaneous lesions in Prader-Willi syndrome]. AB - INTRODUCTION: The Prader-Labhart-Willi syndrome was first described in 1956. Prader-Labhart-Willi syndrome is the most common genetic form of human obesity and the incidence of Prader-Labhart-Willi syndrome has been estimated to 1 in 10,000 or 25,000 live births. Skin-picking was frequently reported in Prader Labhart-Willi syndrome and two patients who displayed repetitive skin picking are described. OBSERVATIONS: Two childrens (6 year-old girl and 7 year-old boy) were examined and noted superficial ulcers of their arms and legs. This cutaneous lesions were induced by children themselves. Skin-picking, in our cases, were associated with behavior problems (temper tantrums, violence). CONCLUSIONS: Skin picking appears to occur in the great majority of patients with Prader-Labhart Willi syndrome and constitutes a minor criteria of diagnosis. Hypopigmentation in Prader-Labhart-Willi syndrome appears to be as common as previously features. Significant differences in hair color, sun sensitivity and complexion were found between those patients with chromosome 15 deletion and those with normal chromosome. Association between obesity (onset before 6 years) and skin picking constitute a sign for diagnosis of Prader-Labhart-Willi syndrome. PMID- 9739897 TI - [Chronic sclerodermiform syndrome disclosing subcutaneous T-cell lymphoma]. AB - INTRODUCTION: Subcutaneous tissue is an uncommon primary localization for T-cell lymphomas. Panniculitis with recurrent papulonodules are reported in most cases. CASE REPORT: We report a case in which small-cell pleiomorphic non-Hodgkin lymphoma developed initially in subcutaneous tissue followed by secondary systemic extension. The first clinical manifestation was a sclerodermiform syndrome involving the four limbs. DISCUSSION: This type of lymphoma appears to have a polymorphic clinical presentation. Physicians should be aware of this type of lymphoma and use immunohistological techniques for early diagnosis. PMID- 9739898 TI - [Enoxaparin-induced cutaneous necrosis localized on insulin lipodystrophies]. AB - INTRODUCTION: Low-molecular weight heparin-induced cutaneous necrosis is exceptional. Pathogenesis remains unclear. We report an exceptional case with elective localization of the necrotic areas in insulin lipodystrophic tissue. CASE REPORT: A 69-year old patient developed areas of skin necrosis after starting enoxaparin therapy. These areas were located far from the points of injection and focalized on skin areas where the patient had been injecting insulin daily for the last four years. These areas had an aspect of insulin lipodystrophy. Biopsy specimens showed leukocytoclastic vasculitis. There were no associated biological anomalies. One month later, prick-tests were made with different low-molecular weight heparins and calcium heparinate in a lipodystrophic area together with an enoxaparin control test in healthy skin. The only positive test was for enoxaparin in an insulin lipodystrophic area (hard erythema at 24 hours). Histology at 72 hours demonstrated leukocytoclastic vasculitis. DISCUSSION: Six cases of cutaneous necrosis induced by low-molecular weight heparin have been reported, including three cases with enoxaparin. Two pathophysiological mechanisms could be involved: (i) localized heparin-dependent platelet aggregation, or (ii) vasculitis induced by type III hypersensitivity reaction. In our case, the leukocytoclastic aspect of the vasculitis was compatible with an immune complex hypersensitivity reaction. The localization of the necrotic areas would be explained by enoxaparin-induced preferential deposit of immune complexes in the vascular turbulences present in lipodystrophic areas. PMID- 9739899 TI - [Oculomotor nerve paralysis with complete ptosis in herpes zoster ophthalmicus: 2 cases]. AB - INTRODUCTION: Only few studies focus on ocular motor paralyses in herpes zoster ophtalmicus. We report 2 cases of complete ptosis resulting from paralysis of the superior lid levator, appearing at day 6 and 7 of an ophtalmic herpes zoster under treatment with acyclovir. CASE REPORTS: Case 1: A 68 year old woman presented an history of ophtalmic herpes zoster with kerato-conjunctivitis and uveitis treated with acyclovir. At the third day of the treatment and 7th day of the ophtalmic zoster, an incomplete paralysis of the oculomotor nerve appeared resulting in a complete ptosis. The treatment was carried on until the 21st day without improvement. Four months later, all symptoms had completely cleared. CASE 2: A 66 year old woman was treated with acyclovir for an ophtalmic herpes zoster without ocular involvement. At the 4th day of the treatment and 6th day of the onset of the ophtalmic zoster, a paralytic ptosis and a acute epithelial keratitis appeared. Acyclovir treatment was continued for 10 days. The ptosis resolved gradually during 2 months. DISCUSSION: The manifestation of a complete ptosis with paralysis of the oculomotor nerve or of one of its branch is rarely seen in ophtalmic herpes zoster. However minor symptoms are often detected when patients were carefully examined with regard to external ocular movements. The physiopathological mechanism are discussed about. The possible action of an early antiviral treatment on the prevention of these complications is not known. In our two cases, a paralytic ptosis broke out suddenly, even under treatment with acyclovir for respectively 3 and 4 days. For future prospective studies about antiviral drugs for ophtalmic herpes zoster, a systematic evaluation of these neurological symptoms would be interesting. PMID- 9739901 TI - [A case for diagnosis: unilateral nevoid telangiectasia]. PMID- 9739900 TI - [Aseptic disseminated abscesses: association with neutrophilic skin diseases and chronic inflammatory bowel diseases]. PMID- 9739902 TI - [Herpes simplex virus: mechanism of latency and reactivation phases]. PMID- 9739903 TI - [Amniotic band syndrome]. PMID- 9739904 TI - [Excision limits and reoperation in cutaneous carcinoma]. PMID- 9739905 TI - [Monthly question: local or systemic treatment: what is your choice?]. PMID- 9739906 TI - [Epstein-Barr virus and cutaneous T lymphoma]. PMID- 9739907 TI - [Comparison of the analgesic efficacy of EMLA 5% cream and lidocaine infiltration for biopsy of the genital mucosa]. AB - OBJECTIVE: To compare the analgesic efficacy of EMLA 5 p. 100 cream versus Xylocaine 1 p. 100 infiltration for biopsies of the genital mucosa. PATIENTS AND METHODS: 63 adult patients were randomized. EMLA (0.3-5 g) was applied during 7 12 minutes, and Xylocaine 1 p. 100 (0.2-5 ml) was infiltrated 0-10 minutes before biopsy. Pain during the anaesthetic procedure and the biopsy was assessed by the patient using a Visual Analogue Scale. RESULTS: Pain scores were significantly lower with EMLA application than Xylocaine infiltration, but infiltration resulted in better surgical anaesthesia. The combined pain scores (anaesthetic procedure and biopsy) were lower in the EMLA group, but this difference failed to reach statistical significance. CONCLUSION: EMLA is a less painful anaesthetic procedure than infiltration, but has a lower analgesic efficacy. EMLA can be used as an alternative to infiltration for biopsies of the genital mucosa. PMID- 9739909 TI - [Hypersensitive urticarial vasculitis after natisedine intake]. AB - BACKGROUND: Various skin and mucosal reactions can be observed after administration of quinidine derivatives. CASE REPORT: A patient who was taking Natisedine (quinidine phenylethyl barbiturate) intermittently and at reintroduction developed a papulopurpuric eruption (without thrombopenia) producing extensive centrifugal annular infiltration and central healing which regressed approximately one week after drug withdrawal. This eruption was associated with moderate 24 h proteinuria. The clinical aspect was that of vasculitis purpura as confirmed histology. Direct immunofluorescence only demonstrated C3 deposits in the vessel walls of the superficial dermis. The quinidine moiety of this drug (currently removed from the formulation) appears to be the responsible agent (imputability score: 13 B3). DISCUSSION: Thrombopenic purpura by synthesis of anti-platelet antibodies induced by quinidine derivatives is well known. Inversely, cases of non-thrombopenic purpura imputable to these same derivatives is uncommon (7 reported cases). The pathophysiological mechanisms involved might be similar (antigenic similarity between the platelet surface and endothelium). PMID- 9739908 TI - [Cell cycle: regulation and abnormalities in epidermal tumors]. AB - The size of a tissue such as the epidermis, and its potential growth, results from the balance between cell production and loss. The mechanisms controlling these two parameters are still poorly understood. Major advances have however been recently achieved in our knowledge of cell cycle regulation and programmed cell death. Dysregulation could explain the development of neoplastic tumors. Basal cell carcinoma basically results from a fall in programmed cell death. Spindle cell carcinoma results from an increased cell production. Both mechanisms are involved in melanoma. One must however keep in mind the fact that we are dealing with hypotheses which remain to be verified. PMID- 9739910 TI - [Diffuse pigmentation (nail, mouth and skin) associated with HIV infection]. AB - INTRODUCTION: Nail dyschromia in patients infected with human immunodeficiency virus (HIV) was first described in 1987 by Furth and Kazakis. It has since been reported in patients with the acquired immunodeficiency syndrome (AIDS), predominantly in patients treated with zidovudine. CASE REPORT: We describe the case of a 37 years old white woman, who developed AIDS in 1994, with nail longitudinal colored bands, oral and cutaneous pigmentation without taking zidovudine. DISCUSSION: There have been four reports of nail pigmentation in HIV infected patients who had no received this antiviral agent. The singularity of our case is the onset in a white woman. A lot of causes must be evoked and biology must be done, with histopathologic study when it is possible. PMID- 9739911 TI - [Gilbert disease and isotretinoin]. AB - INTRODUCTION: Because of the potential hepatotoxicity of retinoids, prescription of isotretinoin is always very carefully made in healthy subjects, and prohibited in case of concomitant hepatopathy. Gilbert's syndrome consists of chronic, mild, unconjugated hyperbilirubinemia. In this syndrome, isotretinoin has been reported twice to be perfectly tolerated, and once even beneficial. We report here a new case of good tolerance and even improvement of a Gilbert's syndrome during isotretinoin therapy. CASE REPORT: A 17-year-old man with Gilbert's syndrome presented with a nodulocystic acne. Topical agents had been inefficient, and cyclines bad tolerated. Thus isotretinoin has been gradually introduced, with a regular monitoring of the liver function. We observed a steady decrease of the bilirubinemia during the course of isotretinoin, and then a reappearance of hyperbilirubinemia as soon as posology was diminished and particularly after completion of isotretinoin therapy. DISCUSSION: A review of the literature finds only very few cases of hepatic injuries caused by isotretinoin, contrary to etretinate. Safety of isotretinoin in Gilbert's syndrome was first observed in 1984, but its beneficial effects have only recently been described by Wang et al., and we report here a similar case. Pharmacological mechanisms remain hypothetic. However, considering the prevalence of Gilbert's syndrome and its usual first expression during postpubertal period, it seems to us interesting for therapeutic practice to know that isotretinoin is not less safe in these patients. PMID- 9739912 TI - [Pemphigoid vegetans. An immunoelectron microscopic study]. AB - INTRODUCTION: Pemphigoid vegetans is a rare disease. It has a clinical resemblance to pemphigus vegetans, but there are histological and immunopathological features of bullous pemphigoid. CASE REPORT: We describe a case in a 57-year-old-man who had developed intertriginous vegetating plaques. Histologic examination of a skin biopsy specimen and direct immunofluorescence microscopy of a biopsy specimen were those of bullous pemphigoid. Immunoblot studies and indirect immunofluorescences of salt-split skin were negative. Direct immunoelectron microscopy was consistent with bullous pemphigoid. DISCUSSION: Only five cases have been reported. We describe the first case including direct immunomicroscopic findings which suggest that pemphigoid vegetans is a subtype of bullous pemphigoid. The other interest was a remarkable improvement with tetracyclines. PMID- 9739913 TI - [Congenital skin defect and fetus papyraceus. A case]. AB - BACKGROUND: Congenital skin defect is an uncommon condition. The term of "congenital skin aplasia" should be avoided since the origin may not necessarily be congenital malformation. The scalp is involved in 80 p. 100 of the cases. CASE REPORT: We observed a congenital skin defect located exclusively on the trunk. The infant lacked wide areas of skin in symmetrical star-like configurations. DISCUSSION: The notion of a twin expulsed after 4 months gestation suggested the diagnosis of congenital skin defect and fetus papyraceus or group V congenital skin aplasia in the Frieden classification as recalled. This diagnosis must not be overlooked as the prognosis is good. Closure with atrophic skin usually occurs within a few weeks. The etiopathogenesis remains obscure. PMID- 9739914 TI - [Pharmaco-surveillance of drug-induced cutaneous accidents]. PMID- 9739915 TI - [A case for diagnosis: atrophodermia vermiculata]. PMID- 9739916 TI - [Vulvar lichen sclerosus]. PMID- 9739917 TI - [Percutaneous absorption: topical and systemic treatments]. PMID- 9739918 TI - [Question of the month: how do you treat buccal erosive lichen?]. PMID- 9739919 TI - [I RPHUSE, I RMR, I treat]. PMID- 9739920 TI - [Delay in diagnosing melanoma. A prospective study in 102 patients]. AB - INTRODUCTION: Knowledge of the causes of melanoma and reasons for diagnosis delay is essential for early management. PATIENTS AND METHODS: One hundred two patients consulting for melanoma at the Saint-Louis Hospital in Paris from January 1, 1994 to December 31, 1995 were asked to respond to a standardized questionnaire. Time to diagnosis and the different time fractions were analyzed by socio-demographic characteristics and by pathology features. RESULTS: Meantime from the first signs of a new lesion or modification of an old lesion to exeresis of melanoma was 20.4 months. Most of the delay prior to diagnosis was patient-related; lack of knowledge about the early clinical signs of melanoma appeared to be the most important cause of delay. Time to diagnosis was not significantly correlated to the thickness of the melanoma. DISCUSSION: Our results are compared with two similar series reported in other countries during the last ten years. The lack of correlation between the thickness of the melanoma and time to diagnosis appears to be explained, at least in part, by the biological variability of melanomas. PMID- 9739921 TI - [Detection of circulating neoplastic cells by reverse transcriptase and polymerase chain reaction in melanoma]. AB - BACKGROUND: Circulating melanocytes can be detected in peripheral blood of patients with malignant melanoma by means of tyrosinase messenger RNA amplification. In this study we especially examined peripheral blood from patients with stage II melanoma before and after lymph node dissection for the detection of these circulating melanoma cells. Indeed the presence of regional nodal metastasis is one of the most important prognostic factors in patients with cutaneous melanoma. PATIENTS AND METHODS: Blood samples were collected from 20 normal patients, 42 patients with stage I melanoma and 23 patients with stage III melanoma. Twenty patients with stage II melanoma were tested 3 days before lymph node dissection and 2 a 8 weeks after. To identify circulating melanocytes, we used coupled reverse-transcription and polymerase chain reaction to target tyrosinase messenger RNA. RESULTS: None of the 20 patients with stage II melanoma had detectable circulating melanocytes before lymph node dissection. By contrast, 7 of them became transiently PCR positive in the 8 weeks following surgery. We observed no evidence of correlation between the presence of circulating melanocytes after lymph node dissection and the risk of relapse within 6 months after surgery or the presence of capsule breaking or the number of involved lymph nodes. Sixty-nine percent of stage III patients and none of stage I patients were found to have circulating melanocytes. DISCUSSION: Our study suggests that melanoma cells could circulate transiently after lymph node dissection. Confrontation of our results with literature data, despite important discrepancies related in part to sensibility technique, shows that the presence of circulating melanoma cells is correlated to the clinical stage. Prognostic value of these circulating cells need to be further assessed by prospective studies with large number of patients and long follow-up. PMID- 9739922 TI - [Methi-resistant Staphylococcus aureus myositis]. AB - INTRODUCTION: Pyomyositis are relatively rare in our countries. CASE REPORT: A 73 year-old-man presented with leg pains and septicemia. Diagnosis of pyomyositis was made and a large incision was performed after which the patient had a progressive improvement. DISCUSSION: Diagnosis of pyomyositis may be difficult in early stages. Diagnosis is greatly facilitated by magnetic resonance imaging. Responsibility of Staphylococcus aureus in cases of pyomyositis due to methi resistant Staphylococcus aureus may be evocated even if patients was not hospitalized. PMID- 9739923 TI - [Paraneoplastic pemphigus with tracheobronchial involvement]. AB - BACKGROUND: Paraneoplastic pemphigus is a bullous skin disease with characteristic polymorphous clinical presentation and precise histological and immunological features. We report a case of paraneoplastic pemphigus associated with chronic lymphoid leukemia involving the tracheobronchial epithelium. CASE REPORT: A patient with chronic lymphoid leukemia developed pluriorificial lesions. There were several conjunctival, buccal and genital erosions associated with erosive plaques on the trunk, Nikolski's sign and bullous lesions suggestive of paraneoplastic pemphigus. Histology examination of a bulla showed intraepidermal blistering and suprabasal acantholysis. Direct immunofluorescence evidenced intercellular IgG and C3 deposits. Search for anti-intercellular substance antibodies was positive with fluorescence on specific paraneoplastic pemphigus substrates. At immunotransfer, the serum recognized several bands corresponding to 250, 230, 210 and 190 kD antigens, confirming the diagnosis of paraneoplastic pemphigus. Several days later, the patient's general condition deteriorated with bronchorrhea. Bronchial endoscopy visualized ulceronecrotic plaques. Tracheal biopsy evidenced acantholytic cells and intraepithelial cleavage. General corticosteroid therapy was initiated and led to improvement of the skin lesions but the patient died rapidly from pneumonia. Autopsy confirmed the presence of epithelial cleavage and acantholysis involving the trachea and bronchi. DISCUSSION: This case illustrates the difficulty of diagnosing paraneoplastic pemphigus in the early stages. The pluriorificial lesions were suggestive of a Stevens-Johnson syndrome. Besides the genital, conjunctival and buccal mucosa, other mucosa can be involved. In our case, despite the absence of an immunological element, histology was highly suggestive of specific tracheobronchial localizations. The presence of such lesions, which should be searched for in all cases with bronchopulmonary manifestations, worsens the prognosis. PMID- 9739924 TI - [Syphilitic labial leucokeratosis]. AB - BACKGROUND: Several diagnoses, including syphilis, can be entertained in patients with leukokeratosis of the buccal mucosa. We report a case of labial leucokeratosis which revealed latent syphilis. CASE REPORT: A 36-year-old man with a past history of genital syphilis chancre which have been treated 12 years earlier, developed buccal leucokeratosis with no other clinical manifestation. Histology showed dermal infiltration containing plasma cells, polynuclears and lymphocytes. Blood tests were positive for syphilis. Complementary examinations were unable to detect another localization. Leucokeratosis regressed completely after one injection of Extencilline. There has been no recurrence at one year. DISCUSSION: The clinical and histological presentations of syphilis can mimic different skin diseases. Serodiagnosis alone is significant. Isolated buccal lesions are rarely described in syphilis suggesting serodiagnosis should always be ordered. Whatever the clinical stage of the diseases, serological surveillance after treatment for syphilis is essential. PMID- 9739925 TI - [Acute mast cell leukemia disclosed by vasomotor flushing]. AB - INTRODUCTION: Acute mast cell leukemia is a rare and severe disease. We report herein a case associated with a flush syndrome. CASE REPORT: A 44-year-old man, presented with a flush of face and trunk. Bone marrow was infiltrated with immature mast cells. In spite of chemotherapy and bone marrow transplantation the patient deceased. DISCUSSION: Pheochromocytoma, carcinoid tumor, and mastocytosis are associated with a flush syndrome. In our patient the diagnosis was an acute mast cell leukemia. Acute mast cell leukemia can follow systemic mastocytosis or occur de novo. This disease is of poor prognosis. No treatment is available. PMID- 9739926 TI - [Congenital eccrine angiomatous hamartoma]. AB - BACKGROUND: Eccrine angiomatous hamartoma is an uncommon skin disease with vascular and sudoral components: less than thirty cases have been reported in the literature. CASE REPORT: A 3.5 month-old female infant developed a painful angiomatous plaque on the abdomen which was first seen 15 days after birth. Histology showed numerous vessels and eccrine sudoral glands in the dermis. Exeresis could not be complete and was followed for 15 years. There has been a slight hyperpigmentation with localized hyperhidrosis over the plaque. This hypersudation confirms the diagnosis which could not be confirmed on the initial biopsies. DISCUSSION: This case of congenital eccrine angiomatous hamartoma could only be diagnosed late in the clinical course. This is the first case report on the abdomen, 80 p. 100 of the previous reports were on the limbs. After a 15 year follow-up, there was no progression of the hamartoma and pain regressed gradually. Without secondary development of localized hyperhidrosis, the diagnosis could not have been made. Careful comparison of pathology findings and clinical expression are essential for the diagnosis of eccrine angiomatous hamartoma. PMID- 9739927 TI - [Anitis, vulvar edema and macrocheilitis disclosing Crohn disease in a child: value of metronidazole]. AB - BACKGROUND: We report a case of oro-genital and perianal Crohn's disease which progressed for 4 years with no digestive involvement in a 12-year-old girl. CASE REPORT: At the age of 8 years, a young girl developed recurrent anal fissures. Voluminous vulvar edema developed at 12 years with fissurar macrocheilitis. There were no digestive signs and the diagnosis of Crohn's disease was obtained on the basis of granulomatous epithelioid infiltration of biopsy specimens (Bauhin valve, anus, vulva). Metronidazole given per os at the dose of 25 mg/kg/day for 6 months led to partial significant symptomatic remission. DISCUSSION: Unilateral or bilateral vulvar edema is highly suggestive of Crohn's disease even if the classical digestive inflammatory signs are absent. Demonstration of perianal lesions (erythema, pseudocondylomatous formations) must not mislead the diagnosis (sexual abuse). Simultaneous granulomatous lesions in the genital and labial regions is exceptional in Crohn's disease. Ileocolonoscopy is indicated in such cases and alone can demonstrate latent digestive inflammatory processes. Different agents have been proposed for treatment of local Crohn's disease skin lesions. It is difficult to evaluate their efficacy due to the spontaneous variability of disease expression. PMID- 9739928 TI - [Vulvar Crohn disease: 3 cases]. AB - BACKGROUND: Vulvar involvement in Crohn's disease is uncommon. The elementary lesion is usually an ulceration: 44 cases have been reported in the literature. In 20 p. 100 of the cases, vulvar involvement is the only manifestation of the disease. We report 3 new cases of vulvar lesions in Crohn's disease. CASE REPORTS: The first case had vulvar lesions which complicated ileocolic Crohn's disease, in the other two cases there were no digestive manifestations. One patient developed voluminous edema of the right labium. The diagnosis of Crohn's disease of the vulva was suggested by the characteristic lesions, presence of epithelioid and giant cell granuloma at histology examination of the vulva specimen and finally on rapidly favorable course after treatment with sulfasalazine. The ulcers regressed with medical treatment but remission was temporary with recurrence at treatment withdrawal. DISCUSSION: These cases underline the difficulties encountered in establishing the diagnosis of Crohn's disease in patients who develop vulvar ulcerations alone with no signs of digestive disease and emphasize the difficult problem of long-term control. PMID- 9739929 TI - [Primary cutaneous monomelic B-cell lymphoma]. AB - BACKGROUND: Cutaneous B cell lymphomas, especially when appearing as a monomelic papulonodular eruption, are rare. PATIENT: Ms H. 87-year-old, consulted for a papulonodular eruption of the left lower limb which developed during the past 5 months. This limb had been the site of a lymphedema since a traumatism 8 years ago. Histopathological analysis and immunostaining of a nodule showed that it was a large cell lymphoma of follicular stem cells. There was no extracutaneous involvement and the patient was successfully treated with radiotherapy. Two months after the completed radiotherapy a cutaneous relapse on the trunk and the upper limbs was treated with cyclophosphamide-vincristine-prednisone chemotherapy. DISCUSSION: Lymphedema probably played a role in the genesis of this lymphoma presumably by reducing the local immune response. It may have harmed endothelial cells and maintained an antigenic stimulation leading first to lymphocyte hyperplasia and eventually to a true lymphoma, in the same way this has been proved for some MALT lymphomas. PMID- 9739930 TI - [A case for diagnosis: multiple tricholeiomyoma]. PMID- 9739931 TI - [A case for diagnosis: palmoplantar lichen]. PMID- 9739932 TI - [Antiretroviral treatment in human immunodeficiency virus infection]. PMID- 9739933 TI - [Can cutaneous intolerance reactions to caryolysine be prevented or controlled?]. PMID- 9739934 TI - [In vitro drug efficacy and fungi isolated from skin samples]. PMID- 9739935 TI - [Evaluation of cosmetics: a challenge for dermatology]. PMID- 9739936 TI - [Contact eczema induced by propylene glycol. Concentration and vehicle adapted for for patch tests]. AB - INTRODUCTION: Contact dermatitis to propylene glycol, a widely used compound, is often difficult to evidence with skin tests. CASE REPORTS: We observed three cases of contact eczema to a dermal cream (Zovirax) used for labial herpes simplex. Patch-tests were positive in all three cases when the entire product was used but negative for each of the constituent components. The initial diagnosis could be an allergic reaction to the composition between the components as has been described elsewhere. Skin tests were completed with patch-tests using propylene glycol at concentrations over 5 p. 100 or with a vehicle other than vaseline (commercial tests use 5 p. 100 propylene glycol in vaseline). The results of these tests provided evidence allowing the diagnosis of contact dermatitis to the dermal cream due to allergic reaction to propylene glycol. DISCUSSION: Our three cases illustrate the frequency of false negative reactions to propylene glycol on commercial patch-tests. In agreement with data in the literature, these tests show that propylene glycol must be used at concentrations up to 10 to 20 p. 100 to identify allergic reactions with patch-tests. PMID- 9739937 TI - [Cutaneous Kaposi disease disclosing acquired immunodeficiency syndrome in a child]. AB - INTRODUCTION: Kaposi's sarcoma associated with acquired immunodeficiency syndrome is uncommon in children and cutaneous localizations are rare. We report a case of pediatric cutaneous Kaposi's sarcoma that revealed a human immunodeficiency virus infection. OBSERVATION: An 8-years-old girl native of the Ivory Coast, with normal statural and psychomotor development, presented cutaneous Kaposi's sarcoma after varicella. The serodiagnosis was positive for type 1 human immunodeficiency virus and CD4 lymphocytes count was 9/mm3. A tonsil localization of Kaposi's sarcoma occurred and bleomycin was a short time effective. A relapse of cutaneous Kaposi's sarcoma with digestive, pulmonary and neurological symptoms was transitorily controlled by the association prednisone-vinblastine-doxorubicin, but death occurred with recurrence of pharyngeal Kaposi's sarcoma. DISCUSSION: The review of literature shows that in pediatric acquired immunodeficiency syndrome, failure to thrive, encephalopathy and opportunistic infections are common. On the other hand, Kaposi's sarcoma is unusual and cutaneous localizations are especially observed when the contamination is postnatal and late. In our case the contamination is presumed perinatal and the human immunodeficiency virus infection was asymptomatic until 8 years old. Kaposi's sarcoma was the cause of the most presenting symptoms and of death, without demonstrated opportunistic infections. PMID- 9739938 TI - [Contact allergy to beta blockaders in eye drops: cross allergy?]. AB - INTRODUCTION: Beta-blockers in eye-drops are widely used for the treatment of glaucoma. The potential allergic effect was only recently recognized. CASE REPORT: A 65-year-old man had been treated with eye-drops containing beta blockers for bilateral chronic glaucoma for 14 years. During the last two years, he developed eczema localized on the upper and lower eyelids. Allergy screening confirmed the implication of timolol and befunolol which had been used successively. Later prescription of eye-drops containing carteolol led to recurrence of the eczema. DISCUSSION: This case of contact allergy with three different beta-blockers in the same patient is similar to others reported in the literature. All beta-blockers have a similar chemical structure, but it cannot act as a haptene. The proposed hypothesis is a cross-sensitivity which develops after primary metabolism to a common aldehyde. The risk of recurrence is high if another beta-blocker eye-drop compound is prescribed in a sensitized patient. The risk of side effects in such sensitized patients when taking oral beta-blockers is unknown. PMID- 9739939 TI - [Waldenstrom's macroglobulinemia with antibasement membrane activity of monoclonal immunoglobulin]. AB - INTRODUCTION: Waldenstrom macroglobulinemia is a rare hematologic disorder with characteristic malignant plasma cell proliferation associated with the secretion of a IgM monoclonal immunoglobulin which is the cause of most of the clinical manifestations. Skin involvement is exceptional. We report a new case of Waldenstrom macroglobulinemia discovered in a patient who developed specific skin lesions with monoclonal immunoglobulin deposits. CASE REPORT: A 50-year-old woman developed spontaneously painful infiltrated erythematopapular plaques over the extension aspects of the limbs. There were no other clinical manifestations. The patient had a kappa IgM monoclonal gammapathy. The diagnosis of Waldenstrom macroglobulinemia was proposed after the discovery of voluminous intra-abdominal adenopathies and was confirmed by immunohistochemistry of the biopsies, indirect immunofluorescence evidenced reactivity against the epidermal basal membrane. Chemotherapy led to successful regression of the skin lesions, the tumoral mass and circulating IgM. DISCUSSION: This is the second reported case of Waldenstrom macroglobulinemia with anti-basal membrane monoclonal immunoglobulin. This IgM would recognize an 82 kD antigen on the dermal side of the dermo-epidermal junction situated in the anchoring fibres of the lamina densa, a zone classically involved in acquired bullous epidermolysis. PMID- 9739940 TI - [Persistent erythema multiforme associated with chronic hepatitis C virus infection. Efficacy of interferon alpha]. AB - INTRODUCTION: Persistant erythema multiforme is a rare form of erythema multiforme with subacute typical and atypical lesions that occur during several months. Some cases are associated with chronic viral infection. CASE REPORT: A 23 year-old man, with a past history of intravenous drug addiction and chronic hepatitis C virus infection, presented persistant erythema multiforme for 18 months. The histopathological picture was those of infectious erythema multiforme and the seric total complement level was low. Two courses of alpha-interferon treatment were quickly efficient on cutaneous lesions, and relapse occurred after discontinuation. DISCUSSION: In previously reported cases of persistant erythema multiforme, etiologic complementary investigations are not always specified. However, viral infections should be considered. In cases of chronic infection, hepatitis C may induce immune disorders through persistent antigenic stimulation. PMID- 9739941 TI - [Caustic dermatitis following application of Pevaryl spray]. PMID- 9739942 TI - [Homotopic response in dermatology]. PMID- 9739943 TI - [A case for diagnosis: herpes gladiatorum]. PMID- 9739944 TI - [A case for diagnosis: cutaneous metastasis of breast neoplasm]. PMID- 9739945 TI - [Cutaneous drug reactions in children]. PMID- 9739946 TI - [Perioral dermatitis in children]. PMID- 9739947 TI - [New antibiotics in 1997: what is their value to the dermatologist?]. PMID- 9739948 TI - [Treatment of venous ulcer of the leg. Recommendations of the Conference of Experts, Oslo 1995]. PMID- 9739949 TI - [Adverse effects of methotrexate: lymphoma and fatigue fractures]. PMID- 9739950 TI - [Question of the month: should herpes zoster in an immunocompetent subject (except ocular herpes zoster) be treated by antiviral agents?]. PMID- 9739951 TI - A piece of my mind. A challenge to licensing boards: the stigma of mental illness. PMID- 9739952 TI - New drug for erectile dysfunction boon for many, "viagravation" for some. PMID- 9739953 TI - Leptin passes safety tests, but effectiveness varies. PMID- 9739954 TI - US military medicine responds to results of terrorism in Africa. PMID- 9739955 TI - First "antisense" drug will treat CMV retinitis. PMID- 9739956 TI - From the Food and Drug Administration. PMID- 9739957 TI - From the Centers for Disease Control and Prevention. Changes in mortality from heart failure--United States, 1980-1995. PMID- 9739958 TI - From the Centers for Disease Control and Prevention. Outbreak of influenza A infection--Alaska and the Yukon Territory, June-July 1998. PMID- 9739959 TI - Antisecretory therapy for bleeding peptic ulcer. PMID- 9739960 TI - Cerebrospinal fluid levels of antiretroviral medications: abstract and commentary. PMID- 9739961 TI - Clinical diagnosis of moles vs melanoma. PMID- 9739962 TI - Clinical diagnosis of moles vs melanoma. PMID- 9739963 TI - Clinical diagnosis of moles vs melanoma. PMID- 9739964 TI - Patients with atrial fibrillation at low risk of stroke. PMID- 9739965 TI - Patients with atrial fibrillation at low risk of stroke. PMID- 9739966 TI - Corporate gifts to academic researchers. PMID- 9739967 TI - Corporate gifts to academic researchers. PMID- 9739968 TI - Managed care for mental health in Tennessee. PMID- 9739969 TI - Managed care for mental health in Tennessee. PMID- 9739970 TI - Sodium reduction and weight loss for elderly patients with hypertension. PMID- 9739971 TI - National autopsy data dropped from the National Center for Health Statistics Database. PMID- 9739972 TI - Oral famciclovir for the suppression of recurrent genital herpes: a randomized controlled trial. Collaborative Famciclovir Genital Herpes Research Group. AB - CONTEXT: Recurrent genital herpes simplex virus (HSV) may be treated episodically, but this may not be sufficient for patients with frequent recurrences. OBJECTIVE: To determine the efficacy and safety of famciclovir in the suppression of recurrent genital HSV infection. DESIGN: A randomized, double blind, placebo-controlled, parallel-group study. SETTING: Thirty university, hospital, or private outpatient referral centers in Canada and Europe. PATIENTS: A total of 455 patients (223 men, 232 women) aged 18 years or older with a history of 6 or more episodes of genital herpes during 12 of the most recent 24 months, in the absence of suppressive therapy, received study medication. INTERVENTION: Oral famciclovir, 125 mg or 250 mg 3 times daily or 250 mg twice daily, or placebo for 52 weeks. MAIN OUTCOME MEASURES: Time to the first recurrence of genital HSV infection; the proportion of patients remaining free of HSV recurrence at 6 months; frequency of adverse events. RESULTS: In an intent-to treat analysis, famciclovir significantly delayed the time to the first recurrence of genital herpes at all dose regimens (hazard ratios, 2.9-3.3; P<.001); median time to recurrence for famciclovir recipients was 222 to 336 days compared with 47 days for placebo recipients. The proportion of patients remaining free of HSV recurrence was approximately 3 times higher in famciclovir recipients (79%-86%) than in placebo recipients (27%) at 6 months (relative risks, 2.9-3.1; P<.001); efficacy was maintained at 12 months. Famciclovir was well tolerated with an adverse experience profile comparable to placebo. CONCLUSIONS: Oral famciclovir (125 mg or 250 mg 3 times daily or 250 mg twice daily) is an effective, well-tolerated treatment for the suppression of genital HSV infection in patients with frequent recurrences. PMID- 9739973 TI - Has the California tobacco control program reduced smoking? AB - CONTEXT: Comprehensive community-wide tobacco control programs are considered appropriate public health approaches to reduce population smoking prevalence. OBJECTIVE: To examine trends in smoking behavior before, during, and after the California Tobacco Control Program. DESIGN: Per capita cigarette consumption data (1983-1997) were derived from tobacco industry sales figures. Adult (> or =18 years) smoking prevalence data were obtained from the National Health Interview Surveys (1978-1994), the California Tobacco Surveys (1990-1996), the Current Population Surveys (1992-1996), and the California Behavioral Risk Factor Survey and its supplement (1991-1997). Trends were compared before and after introduction of the program, with the period after the program being divided into 2 parts (early, 1989-1993; late, 1994-1996). MAIN OUTCOME MEASURES: Change in cigarette consumption and smoking prevalence in California compared with the rest of the United States. RESULTS: Per capita cigarette consumption declined 52% faster in California in the early period than previously (from 9.7 packs per person per month at the beginning of the program to 6.5 packs per person per month in 1993), and the decline was significantly greater in California than in the rest of the United States (P<.001). In the late period, the decline in California slowed to 28% of the early program so that in 1996 an average of 6.0 packs per person per month were consumed. No decline occurred in the rest of the United States, and in 1996, 10.5 packs per person per month were consumed. Smoking prevalence showed a similar pattern, but in the late period, there was no significant decline in prevalence in either California or the rest of the United States. In 1996, smoking prevalence was 18.0% in California and 22.4% in the rest of the United States. CONCLUSIONS: The initial effect of the program to reduce smoking in California did not persist. Possible reasons include reduced program funding, increased tobacco industry expenditures for advertising and promotion, and industry pricing and political activities. The question remains how the public health community can modify the program to regain its original momentum. PMID- 9739974 TI - Physicians' experiences and beliefs regarding informal consultation. AB - CONTEXT: Efforts to control medical expenses by emphasizing primary care and limiting specialty care may influence how physicians use informal or "curbside" consultation. OBJECTIVE: To understand physicians' use of and beliefs about informal consultation. DESIGN: Survey mailed in July 1997. PARTICIPANTS: Of a random sample of Massachusetts general internists, pediatricians, cardiologists, orthopedic surgeons (n=300 each), and infectious disease specialists (n=200) surveyed, 1225 were eligible and 705 (58%) responded. MAIN OUTCOME MEASURES: Self reported use of and beliefs about informal consultation. RESULTS: Generalist physicians requested more informal consultations than specialists (median, 3 vs 1 per week; P<.001) and were asked to provide fewer (2 vs 5 per week; P<.001). In multivariate analyses, physicians in a health maintenance organization, multispecialty group, or single-specialty group requested more informal consultations than those in solo practice (82%, 40%, and 28% more, respectively; all P<.001) and were more often asked to provide them (43%, 63%, and 14% more, respectively; all P<.05). Physicians with at least 30% of their income from capitation requested 38% more and were asked to provide 46% more informal consultations than those with little or no income from capitation (both P<.001). Generalists' overall approval of informal consultation was greater than specialists' (mean 5.9 vs 5.1 on a 7-point Likert scale; P<.001), and approval was strongly associated with beliefs about how informal consultation affects quality of care (P<.001). CONCLUSIONS: Use of informal consultation is common, varies by specialty, practice setting, and capitation, and therefore may increase with current trends toward group practice and managed care. Because overall approval of informal consultation is strongly associated with beliefs about how it affects quality of care, this issue should be carefully considered by physicians who participate in informal consultation. PMID- 9739975 TI - Curbside consultation practices and attitudes among primary care physicians and medical subspecialists. AB - CONTEXT: Informal (curbside) consultations are an integral part of medical culture and may be of great value to patients and primary care physicians. However, little is known about physicians' behavior or attitudes toward curbside consultation. OBJECTIVE: To describe and compare curbside consultation practices and attitudes among primary care physicians and medical subspecialists. DESIGN: Survey mailed in June 1997. PARTICIPANTS: Of 286 primary care physicians and 252 subspecialists practicing in Rhode Island, 213 primary care physicians and 200 subspecialists responded (response rate, 76.8%). MAIN OUTCOME MEASURES: Self reported practices of, reasons for, and attitudes about curbside consultation. RESULTS: Of primary care physicians, 70.4% (150/213) and 87.5% (175/200) of subspecialists reported participating in at least 1 curbside consultation during the previous week. In the previous week, primary care physicians obtained 3.2 curbside consultations, whereas subspecialists received 3.6 requests for curbside consultations. Subspecialties most frequently involved in curbside consultations were cardiology, gastroenterology, and infectious diseases; subspecialties that were requested to provide curbside consultations more often than they were formally consulted were endocrinology, infectious diseases, and rheumatology. Curbside consultations were most often used to select appropriate diagnostic tests and treatment plans and to determine the need for formal consultation. Subspecialists perceived more often than primary care physicians that information communicated in curbside consultations was insufficient (80.2% vs 49.8%; P<.001) and that important clinical detail was not described (77.6% vs 43.5%; P<.001). More subspecialists than primary care physicians felt that curbside consultations were essential for maintaining good relationships with other physicians (77.2% vs 38.6%; P<.001). CONCLUSIONS: Curbside consultation serves important functions in the practice of medicine. Despite the widespread use of curbside consultation, disagreement exists between primary care physicians and subspecialists as to the role of curbside consultation and the quality of the information exchanged. PMID- 9739976 TI - Increased cancer mortality following a history of nonmelanoma skin cancer. AB - CONTEXT: Cancer registries have reported an increased incidence of melanoma and certain noncutaneous cancers following nonmelanoma skin cancer (NMSC). Whether these findings were attributable to intensified surveillance, shared risk factors, or increased cancer susceptibility remains unclear. OBJECTIVE: To determine whether a history of NMSC predicts cancer mortality. DESIGN: Prospective cohort with 12-year mortality follow-up adjusted for multiple risk factors. SETTING: Cancer Prevention Study II, United States and Puerto Rico. PARTICIPANTS: Nearly 1.1 million adult volunteers who completed a baseline questionnaire in 1982. MAIN OUTCOME MEASURE: Deaths due to all cancers and common cancers. RESULTS: After adjusting for age, race, education, smoking, obesity, alcohol use, and other conventional risk factors, a baseline history of NMSC was associated with increased total cancer mortality (men's relative risk [RR], 1.30; 95% confidence interval [CI], 1.23-1.36; women's RR, 1.26; 95% CI, 1.17-1.35). Exclusion of deaths due to melanoma reduced these RRs only slightly. Mortality was increased for the following cancers: melanoma (RR, 3.36 in men, 3.52 in women); pharynx (RR, 2.77 in men, 2.81 in women); lung (RR, 1.37 in men, 1.46 in women); non-Hodgkin lymphoma (RR, 1.32 in men, 1.50 in women); in men only, salivary glands (RR, 2.96), prostate (RR, 1.28), testis (RR, 12.7), urinary bladder (RR, 1.41), and leukemia (RR, 1.37); and in women only, breast (RR, 1.34). All-cause mortality was slightly increased (adjusted men's RR, 1.03 [95% CI, 1.00-1.06]; women's RR, 1.04 [95% CI, 1.00-1.09]). CONCLUSIONS: Persons with a history of NMSC are at increased risk of cancer mortality. Although the biological mechanisms are unknown, a history of NMSC should increase the clinician's alertness for certain noncutaneous cancers as well as melanoma. PMID- 9739977 TI - Exercise echocardiography or exercise SPECT imaging? A meta-analysis of diagnostic test performance. AB - CONTEXT: Cardiac imaging has advanced rapidly, providing clinicians with several choices for evaluating patients with suspected coronary artery disease, but few studies compare modalities directly. OBJECTIVES: To review the contemporary literature and to compare the diagnostic performance of exercise echocardiography (ECHO) and exercise single-photon emission computed tomography (SPECT) imaging in the diagnosis of coronary artery disease. DATA SOURCES: Studies published between January 1990 and October 1997 identified from MEDLINE search; bibliographies of reviews and original articles; and suggestions from experts in each area. STUDY SELECTION: Articles were included if they discussed exercise ECHO and/or exercise SPECT imaging with thallous chloride TI 201 (thallium) or technetium Tc 99m sestamibi for detection and/or evaluation of coronary artery disease, if data on coronary angiography were presented as the reference test, and if the absolute numbers of true-positive, false-negative, true-negative, and false-positive observations were available or derivable from the data presented. Studies performed exclusively in patients after myocardial infarction, after percutaneous transluminal coronary angioplasty, after coronary artery bypass grafting, or with recent unstable coronary syndromes were excluded. DATA EXTRACTION: Clinical variables, technical factors, and test performance were independently extracted by 2 reviewers on a standardized spreadsheet. Discrepancies were resolved by consensus. RESULTS: Forty-four articles met inclusion criteria. In pooled data weighted by the sample size of each study, exercise ECHO had a sensitivity of 85% (95% confidence interval [CI], 83%-87%) with a specificity of 77% (95% CI, 74% 80%). Exercise SPECT yielded a similar sensitivity of 87% (95% CI, 86%-88%) but a lower specificity of 64% (95% CI, 60%-68%). In a summary receiver operating characteristic model comparing exercise ECHO performance to exercise SPECT, exercise ECHO was associated with significantly better discriminatory power (parameter estimate, 1.18; 95% CI, 0.71-1.65), when adjusted for age, publication year, and a setting including known coronary artery disease for SPECT studies. In models comparing the discriminatory abilities of exercise ECHO and exercise SPECT vs exercise testing without imaging, both ECHO and SPECT performed significantly better than exercise testing. The incremental improvement in performance was greater for ECHO (3.43; 95% CI, 2.74-4.11) than for SPECT (1.49; 95% CI, 0.91 2.08). CONCLUSIONS: Exercise ECHO and exercise SPECT have similar sensitivities for the detection of coronary artery disease, but exercise ECHO has better specificity and, therefore, higher overall discriminatory capabilities as used in contemporary practice. PMID- 9739978 TI - Ability to obtain medical care for the uninsured: how much does it vary across communities? AB - CONTEXT: Communities differ in the way that medical care for medically indigent persons is organized and delivered, which is likely to result in differences across communities in the ability of uninsured persons to obtain medical care. Changes in the health care system, many of which are driven locally, may further exacerbate these differences. OBJECTIVE: To examine the extent of variation across US communities in the ability of uninsured persons to obtain medical care and the extent to which health status and other characteristics of the uninsured population account for these differences. DESIGN: Analysis of the 1996-1997 Community Tracking Study Household Survey. SETTING: A nationally representative sample of the US civilian, noninstitutionalized population residing in 60 randomly selected communities. Larger sample sizes were obtained for 12 of these communities, which were randomly selected to represent metropolitan areas in the United States with more than 200000 persons. PARTICIPANTS: A total of 60 446 individuals and 7200 uninsured persons. MAIN OUTCOME MEASURES: The percentage of persons who either did not obtain needed medical care in the previous year or postponed receiving needed medical care in the previous year. RESULTS: Differences between communities with the highest percentage of uninsured persons reporting difficulty obtaining care and communities with the lowest percentage were more than 2-fold (41.4% vs 18.5%, P<.05). Little of the variation across communities is accounted for by differences in health status or sociodemographic characteristics of the uninsured population. The pattern of variation across communities in the ability of uninsured persons to obtain medical care is not correlated with variations in the ability of privately insured persons to obtain care (Pearson r, 0.04). Simulation results indicate that expanding private or public insurance coverage would not only increase the ability of uninsured persons to obtain medical care but would also reduce the variation across communities. CONCLUSIONS: If people are uninsured, where they live is an important factor in determining the difficulty they have in obtaining care. This is likely to persist given that care for uninsured persons is driven largely by state and local policy, and health system changes are constraining clinicians' ability and willingness to serve uninsured persons in many parts of the country. PMID- 9739979 TI - Long-term suppression of genital herpes. PMID- 9739980 TI - Curbside consultations and the viaduct effect. PMID- 9739981 TI - Marginal medicine. PMID- 9739982 TI - JAMA patient page: sexually transmitted diseases. PMID- 9739983 TI - Cultured human monocytes release proinflammatory cytokines in response to myelin basic protein. AB - In human cultured monocytes we examined the ability of myelin basic protein (MBP) to induce the production of proinflammatory cytokines potentially involved in inflammatory demyelination. Northern blots and specific immunoassays demonstrated that monocytes incubated with optimal doses of MBP showed increased mRNA expression and release of tumor necrosis factor (TNF-alpha), interleukin-1beta (IL-1beta), interleukin-6 (IL-6), interleukin-8 (IL-8) but not of interleukin 12/p40 (IL-12/p40). We also showed that cytokine production by MBP-stimulated monocytes was abrogated by incubation with Dexamethasone. These data suggest that interaction of mononuclear phagocytes with MBP may participate in the regulatory process of cytokine production during inflammatory demyelination and support the beneficial role of corticosteroids therapy in aberrant immune responses to the myelin sheath. PMID- 9739984 TI - Postsynaptic phospholipase C activity is required for the induction of homosynaptic long-term depression in rat hippocampus. AB - The necessity for phospholipase C (PLC), the enzyme which produces the second messenger molecules inositol trisphosphate (IP3) and diacylglycerol, for the induction of long-term depression (LTD) was tested at Schaffer collateral-CA1 synapses in hippocampal slices in vitro. We report here that bath application of a selective cell-permeant PLC inhibitor, U-73122 (10 microM), does block the induction of LTD. In contrast, neither the inactive analog U-73343 (10 microM), nor application of U-73122 during the maintenance phase of LTD, impaired expression of LTD. Furthermore, postsynaptic infusion of U-73122 (100 microM) into single CA1 pyramidal neurons also prevented the induction of LTD. Since mGluR5 is the only metabotropic glutamate receptor subtype coupled to inositide turnover in field CA1, we conclude that the postsynaptic calcium store necessary for the induction of homosynaptic LTD is gated by IP3, through activation of mGluR5 coupled to phospholipase C. PMID- 9739985 TI - Decreased transcription factor junD in brains of patients with Down syndrome. AB - JunD is a member of the Jun family of transcription factors (TF), recently shown to negatively regulate cell growth and antagonizes transformation by the protooncogene ras: c-jun decreases while junD is accumulating when fibroblasts become quiescent. Furthermore, overexpression of junD resulted in slower growth and an increase in cells in G0/G1. Performing gene hunting on fetal Down syndrome (DS) brain we found a sequence downregulated and homologous to junD. This observation made us examine junD protein levels in adult brain specimens. Western blot experiments were carried out in five brain regions of aged patients with DS (n = 9), controls (n = 9) and patients with Alzheimer's disease (AD, n = 9). We found that junD in AD brains were comparable to controls, whereas junD levels were significantly and remarkably reduced in frontal, temporal lobe and cerebellum of patients with DS. These findings may indicate a specific finding in DS and were not linked to the AD-like-neuropathological changes of plaques and tangles, observed in DS from the fourth decade, which is also suggested by the findings of downregulated junD at the mRNA level revealed by the gene hunting technique (subtractive hybridization) in fetal DS brain. We propose that junD plays a role for the impaired development and wiring of DS brain, maybe already early in life. PMID- 9739986 TI - Adenosine protects chick embryonic sympathetic neurons from apoptotic death by 2' deoxyadenosine--importance of ATP in apoptosis. AB - Our past work on nucleoside toxicity in sympathetic neurons has clearly revealed that adenosine and 2'-deoxyadenosine (dAdo) have different mechanisms of action in inducing apoptotic death. For example, adenosine is toxic to neurons only during early phase of growth whereas dAdo kills even mature neurons. In this study, we hypothesize that dAdo-induced apoptosis is initiated when ATP concentration of sympathetic neurons decreases below a critical level. To prove our hypothesis we used adenosine as a tool to replenish ATP levels of sympathetic neurons. We demonstrate that dAdo toxicity in mature sympathetic neurons was fully prevented by adenosine treatment. Furthermore, we demonstrate that depletion of ATP caused by dAdo was prevented by pretreatment with adenosine. These data suggest that intracellular accumulation of adenosine could play a neuroprotective role in preventing death associated with reduction in neuronal ATP concentration. PMID- 9739987 TI - Postural effects of unilateral blockade of glutamatergic neurotransmission in the subthalamic nucleus on haloperidol-induced akinesia in rats. AB - The present study examined the postural effects of the local application of glutamatergic antagonists unilaterally into the subthalamic nucleus (STN), on haloperidol-induced akinesia in rats. After intracerebral injections of MK-801, a selective antagonist of N-methyl-D-aspartate (NMDA) receptor, 6-cyano-7 nitroquinoxaline-2,3-dione (CNQX) disodium, a selective alpha-amino-3-hydroxy-5 methyl-4-isoxazole propionate (AMPA) receptor antagonist, or vehicle, unilaterally into the STN, haloperidol was administered systemically and the elicited behaviors were assessed quantitatively. In rats which received injections of MK-801 or CNQX, but not vehicle, unilaterally into the STN, the administration of haloperidol induced contraversive dystonic posturing. The severity of the deviated posturing was dose-dependent. The present findings revealed that the overactivity of the STN under conditions of dopamine blockade is suppressed by interruptions of glutamatergic inputs, mediated via both NMDA or AMPA receptors, to the STN. Therefore, the present study may provide functional evidence in support of a recently proposed hypothesis, that not only disinhibition from the inhibitory globus pallidus efferents but also excitatory glutamatergic inputs to the STN actually contribute to the overactivity of the STN under dopamine-depleted conditions. PMID- 9739988 TI - Self-regulation of slow cortical potentials in completely paralyzed human patients. AB - The study was intended to answer the question whether self-regulation of brain activity can be operantly learnt when the brain is disconnected from motor periphery. Two neurological patients with nearly complete motor paralysis learned bi-directional control of their slow cortical potentials (SCP) at vertex. After 4 6 weeks training both patients could reliably differentiate between SCP shifts in a negative versus positive direction. With one patient, training has been continued for a subsequent 4 months, which resulted in precise self-control, i.e. the patient was able to produce positive SCP shifts on command with an accuracy of about 95%. This indicates that self-regulation of cortical excitability (as manifested in the SCP) does not require feedback loops from the periphery. Although we cannot rule out that healthy subjects may employ behavioral strategies such as muscle contractions or changes in breathing, obviously humans can also control their SCP without using these strategies. PMID- 9739989 TI - Effects of delta-aminolaevulinic acid upon electrical activity of rat spinal cord in vivo. AB - Bath applied delta-aminolaevulinic acid (ALA, 1-1000 microM), significantly depressed the frequency of spontaneous antidromic activity in the lumbar dorsal roots, and the amplitude of the monosynaptic component of the dorsal root-evoked ventral root reflex, in isolated rat spinal cord in a concentration-related manner. In contrast bath concentrations up to 1 mM ALA were found to produce no significant change in either the conducted afferent volley, nor field potentials recorded in the lumbar dorsal horn. These results indicate that the raised levels of ALA found in cerebrospinal fluid during porphyrias and lead poisoning may contribute to the neurological symptoms of these diseases. PMID- 9739990 TI - Shift type and season affect adaptation of the 6-sulphatoxymelatonin rhythm in offshore oil rig workers. AB - Previously we have shown that the 6-sulphatoxymelatonin rhythm of oil rig workers on a 2-week night shift (1800-0600 h) adapts to the shift via a phase delay. We now report the findings of a study on two offshore drill crews working a 1 week day (1200-0000 h), 1 week night (0000-1200 h) swing shift. Urine samples were collected every 2-3 h throughout the subjective days, with over-sleep collections, for the measurement of 6-sulphatoxymelatonin by radioimmunoassay. One crew (n = 11), studied in November, showed no change in their 6 sulphatoxymelatonin rhythm during night shift. The other crew (n = 7), studied in March, showed a significant phase advance of the rhythm during night shift. The data indicate that both the type of shift and the season influence the direction and degree of adaptation. PMID- 9739991 TI - Hippocampal slices from long-term streptozotocin-injected rats are prone to epileptiform responses. AB - Evoked field potentials were recorded in the CA3, CA1 and dentate gyrus (DG) regions of hippocampal slices from rats injected with streptozotocin (STZ; 60 mg/kg i.p.), to detect whether STZ-induced diabetes entails changes in hippocampal excitability. No change in hippocampal responsiveness was observed in slices from diabetic rats, up to 3 weeks post-STZ. Repetitive population spikes (PSs) reminiscent of an epileptiform hyperexcitability were, however, recorded in CA3 > CA1 > DG areas after more than 4 weeks ('long-term') post-STZ, although the maximal amplitudes were not different in STZ-diabetic versus control rats. Intracellular recordings on CA3 pyramidal neurons confirmed that fimbrial stimulation evokes significantly more action potentials in neurons from 'long term' STZ-diabetic versus control rats. This is the first report of the appearance of repetitive hippocampal responses, particularly in the seizure-prone CA3 area, as a long-term consequence of hyperglycemic STZ treatment in rat. PMID- 9739992 TI - The effects of muscular fatigue on the coordination of a multijoint movement in human. AB - The purpose of the present experiment was to investigate the adaptation to fatigue in a multijoint movement. The subjects' task was to throw a ball towards one of three targets in two conditions of no fatigue and fatigue. Results showed that the number of successful trials decreased with fatigue. Analysis of these successful trials showed that, without fatigue, the final hand velocity resulted from a 'summation of speed' principle from the elbow to the hand joint. With fatigue, a new inter-segmental organization was necessary in order to maintain a good motor performance. This compensating strategy was characterized by the absence of a temporal delay between the elbow and hand peak velocity suggesting that, with fatigue, movement organization was similar to that of a rigid system. In other words, the 'summation of speeds' principle was no more respected. PMID- 9739993 TI - FUSE-binding protein is developmentally regulated and is highly expressed in mouse and chicken embryonic brain. AB - GAP-43 modulates axon guidance and neuronal plasticity. In vitro, FUSE-binding protein (FBP) binds to a segment of GAP-43 mRNA which regulates the stability of the transcript. FBP has also been shown to bind to a c-myc cis element and regulate transcription. In the current work, analysis of RNA and protein expression indicated that FBP is expressed in a distinct spatial temporal pattern during embryonic development. Expression was particularly high in the brain. In the adult, expression was not detected in most tissues but was still prominent in the brain and teste. This finding is consistent with a dual role of the protein as a single-strand polynucleotide-binding protein. PMID- 9739994 TI - Expression of the ataxia-telangiectasia gene (ATM) product in human cerebellar neurons during development. AB - Ataxia-telangiectasia (A-T) is a hereditary disorder, exhibiting progressive cerebellar ataxia. We investigated the expression of the ATM protein in the human CNS. By western blotting, the ATM protein was detected in the cerebellar cortex, but not in the cerebral cortex, at the late gestational stage. Immunohistochemistry revealed that cerebellar neurons, particularly Purkinje cells, were markedly immunoreactive during late prenatal and early postnatal periods, followed by persistent and moderate reactivity in Purkinje cells. The ATM protein was distributed within the cytoplasm of Purkinje cells, but not within the nuclei. The ATM protein seems to play a role as a cytoplasmic protein in neurons of the cerebellar cortex. PMID- 9739995 TI - Upregulation of vascular endothelial growth factor in severe chronic brain hypoxia of the rat. AB - The vascular endothelial growth factor (VEGF) has been shown to be upregulated in acute hypoxia. Although an increase in blood vessel number has been described in severe chronic brain hypoxia, it is unclear whether VEGF is upregulated in this condition. We therefore investigated male inbred Wistar rats, which were exposed for 9 to 13 weeks to decreasing amounts of oxygen, down to 7% O2 (15%: 15 days; 12%, 10%, respectively; 8%: 1 day, 3 weeks, respectively; 7%: 4 weeks). The expression of VEGF was studied by Northern analysis and in situ hybridization in frozen sections of cerebral cortex, hippocampus and cerebellum in six chronic hypoxic and two control rats. We found a marked upregulation of VEGF mRNA in all brain regions investigated, being strongest in cerebral cortex and cerebellum. Our results suggest a potential role of VEGF for vascular growth and vascular permeability observed in chronic cerebral hypoxia. PMID- 9739996 TI - A novel Na+/Ca2+ channel blocker NS-7 inhibits evoked but not spontaneous dopamine release from rat striatum, as measured by intracerebral microdialysis. AB - The spontaneous dopamine release from rat striatum, measured by intracerebral microdialysis, was markedly reduced by local perfusion of tetrodotoxin (1 microM) through the dialysis probe. In addition, striatal microinjection of omega conotoxin GVIA (10 pmol) or omega-agatoxin i.v.A (1 pmol), but not local perfusion of nimodipine, suppressed the spontaneous dopamine release. Therefore, the spontaneous dopamine release may depend on the activity of both Na+ channel as well as N-type and P/Q-type Ca2+ channels. In contrast, local perfusion of a novel Na+- and Ca2+-channel blocker NS-7 (10 microM) did not affect spontaneous dopamine release, whereas it markedly blocked KCl- and veratridine-evoked dopamine release. Therefore, NS-7 may block Na+- and Ca2+-channels only when the ion channels are highly activated. PMID- 9739997 TI - Inhibition of adenosine kinase during oxygen-glucose deprivation in rat cortical neuronal cultures. AB - Adenosine kinase (AK) inhibitors potentiate the actions of endogenous adenosine (ADO) and ameliorate cerebral ischemic damage in animal models. The present study examined the effects of the AK inhibitor, 5-iodotubercidin (5-IT) in an in vitro model of neuronal ischemia, specifically, combined oxygen-glucose deprivation of rat cortical mixed neuronal-glial cultures. Oxygen-glucose deprivation caused extensive neuronal loss which was accompanied by a marked increase in ADO release into the extracellular medium, was ameliorated by exogenous ADO (10 microM(-1) mM), and was exacerbated by a high concentration of the selective A1 receptor antagonist 8-cyclopentyl-1,3-dimethylxanthine (CPT; 10 microM). 5-IT (1 microM) had no effect on extracellular ADO levels nor on neuronal loss. However, AK activity in these cultures was markedly suppressed during oxygen-glucose deprivation. Taken together, these data demonstrate a marked down-regulation of AK activity during oxygen-glucose deprivation in this in vitro model, providing an endogenous mechanism contributing to the accumulation of extracellular ADO, which exerts neuroprotective effects by activating the ADO A1 receptor. PMID- 9739998 TI - Brain derived neurotrophic factor induction of N-methyl-D-aspartate receptor subunit NR2A expression in cultured rat cortical neurons. AB - N-methyl-D-aspartate (NMDA) receptor subunit expression changes during development and following injury in several brain regions. These changes may be mediated by neurotrophic factors, such as brain derived neurotrophic factor (BDNF). Exposure of cultured cortical neurons to BDNF (100 ng/ml) for 24 h produced a significant decrease in the NMDA-induced whole-cell currents sensitive to the NR2B subunit selective NMDA receptor antagonist, CP-101,606, suggesting a relative decrease in NR2B subunit expression. There was a significant increase in NR2A by Western blot analysis. Consistent with the electrophysiology and Western blot analysis, reverse transcriptase-polymerase chain reaction (RT-PCR) amplification revealed that BDNF caused a significant increase in relative NR2A subunit expression, a significant decrease in relative NR2B subunit expression and no change in relative NR2C subunit expression. These results suggest that BDNF enhances NMDA receptor maturation, warranting further study of the mechanism of BDNF effects on NMDA receptor subunit expression and the role these effects play in development and neuronal injury. PMID- 9739999 TI - Induction of tyrosine hydroxylase gene expression in human foetal cerebral cortex. AB - Human foetal cerebral cortex (9-14 weeks gestational age) was dissected out and cultured in microwell plates. It was then treated with brain-derived neurotrophic factor (BDNF, 50 ng/ml), dopamine (10 mM) or their combination. After 5 weeks of this treatment tyrosine hydroxylase (TH)-immunopositive neurones were detected at a level of 0.73% of total neurones present. This represented 300-500 TH + neurones per microwell. None were seen in untreated cultures. This correlates with induction of the entire dopaminergic phenotype in foetal rat cerebral cortex (E1214) by the same co-treatment applied for a much shorter time period (7 days), which implies that the complete dopaminergic phenotype is also induced in cultured human foetal tissue over a longer period, reflecting the 5-fold longer neuronal gestational period. PMID- 9740000 TI - Inhibition of bacterial adherence to a high-water-content polymer by a water soluble, nonsteroidal, anti-inflammatory drug. AB - Deposition and aggregation of lachrymal proteins on the contact lens surface can promote bacterial adherence. Lysozyme is the major tear protein and is also mainly responsible for the formation of protein deposits on contact lenses. Nonsteroidal anti-inflammatory drugs (NSAID) prevent protein aggregation. The effect of a water-soluble NSAID drug on bacterial adherence to high-water content/ionic disposable contact lenses was examined in a radiolabeling study. Dose-related inhibition of adherence of Staphylococcus aureus, S. epidermidis, and Pseudomonas aeruginosa on both pretreated lenses and after adding the drug to the medium was investigated. When the drug was added to the media, maximal inhibition of S. aureus adherence was observed in trypticase soy broth (59-98% at the lower and higher drug concentrations, respectively); inhibition progressively decreased in calf aqueous humor (48-75%), lysozyme (34-63%), and saline (12-20%) solutions. Inhibition of adherence varied with the three bacterial species; it was maximal with S. aureus, intermediate with S. epidermidis, and minimal with P. aeruginosa. When lenses were pretreated with the drug, consistent, and even higher, inhibitory effects were observed. The results suggest that water-soluble NSAIDs could be used in preventive treatments for conjunctivae and corneal infections in contact lens wearers, and may provide a clue as to which compounds might inhibit protein interaction and bacterial adhesion. PMID- 9740001 TI - Viscoelastic behavior of composite ligament prostheses. AB - Despite the compelling need for artificial connective tissue replacements for orthopedic applications, to date, there is no material which can adequately reproduce the mechanical behavior of natural tissue with necessary long-term endurance. In this work, we introduce a novel soft composite material as a more suitable candidate for connective tissue replacement. The material proposed is based on a hydrogel-polymer matrix reinforced with poly(ethylene terephthalate) fibers wound helically to mimic the architecture of the collagen fibers in natural tissue. Macroscopic behaviors such as static stress-strain, stress relaxation, and dynamic frequency responses can be modulated with choice of the components and design of the composite structure. In doing so, the mechanical characteristics of natural ligaments can be qualitatively reproduced and sustained over time. PMID- 9740002 TI - Patterned glass surfaces direct cell adhesion and process outgrowth of primary neurons of the central nervous system. AB - Glass surfaces were patterned with cell-adhesive regions of laminin adhesive peptides YIGSR, RGD, and IKVAV, and cell-repulsive regions of poly(ethylene glycol) (PEG). The patterns were created by sputter-coating titanium and then gold onto glass coverslips through electron microscope grids. Gold surfaces were modified with cysteine-terminated peptides to have approximately 450 fmol/ cm2 of peptide incorporated on the glass coverslips as determined with radiolabeled CGYIGSR. Amine-functionalized glass coverslips were prepared using an amine functionalized silane and then further modified with PEG-aldehyde by a Schiff base reduction. All surfaces were characterized by X-ray photoelectron spectroscopy and water contact angles. Hippocampal neurons, plated from a serum free medium, adhered preferentially to peptide-functionalized surfaces over PEG modified surfaces. Cell adhesion and neurite outgrowth were limited to the peptide region, demonstrating that neurite outgrowth could be directed by a combination of cell-adhesive and cell-repulsive cues. PMID- 9740003 TI - Osseointegration of surface-blasted implants made of titanium alloy and cobalt chromium alloy in a rabbit intramedullary model. AB - The purpose of this study was to compare the osseointegration of surface-blasted Ti6A14V and CoCr implants in vivo. Ti6A14V and CoCr rods blasted with 710 microm A12O3 particles were bilaterally press-fit into the medullary space of distal femora of 24 rabbits. Evaluation was made radiographically, histologically, histomorphometrically (3, 6, and 12 weeks after implantation), and mechanically (12 weeks). Both Ti6A14V and CoCr implants demonstrated good biocompatibility radiographically and histologically. Toluidine blue-stained sections revealed an osteoconductive effect of the blasted surface, and fluorochrome labeling analysis showed active bone formation at the bone-implant interface at as late as 12 weeks for both specimens. CoCr showed significantly lower interfacial shear strength than Ti6A14V although the bone contact area with the implant surface was comparable and no intervening soft tissue at the bone-implant interface could be seen for either implant by scanning electron microscopy backscatter analysis. Unmineralized tissue (cartilage and osteoid) was observed more frequently on the CoCr surface than on the Ti6A14V surface. These data show less osseointegration of CoCr implants with this blasted surface for this short period, possibly due to a slight difference in surface roughness and some negative effects of CoCr on bone attachment. PMID- 9740004 TI - Inhibition of aortic wall calcification in bioprosthetic heart valves by ethanol pretreatment: biochemical and biophysical mechanisms. AB - The effectiveness of ethanol pretreatment on preventing calcification of glutaraldehyde-fixed porcine aortic bioprosthetic heart valve (BPHV) cusps was previously demonstrated, and the mechanism of action of ethanol was attributed in part to both lipid removal and a specific collagen conformational change. In the present work, the effect of ethanol pretreatment on BPHV aortic wall calcification was investigated using both rat subdermal and sheep circulatory implants. Ethanol pretreatment significantly inhibited calcification of BPHV aortic wall, but with less than complete inhibition. The maximum inhibition of calcification of BPHV aortic wall was achieved using an 80% ethanol pretreatment; calcium levels were 71.80+/-8.45 microg/mg with 80% ethanol pretreatment compared to the control calcium level of 129.90+/-7.24 microg/mg (p = 0.001). Increasing the duration of ethanol exposure did not significantly improve the inhibitory effect of ethanol on aortic wall calcification. In the sheep circulatory implants, ethanol pretreatment partly prevented BPHV aortic wall calcification with a calcium level of 28.02+/-4.42 microg/mg compared to the control calcium level of 56.35+/-6.14 microg/mg (p = 0.004). Infrared spectroscopy (ATR-FTIR) studies of ethanol-pretreated BPHV aortic wall (vs. control) demonstrated a significant change in protein structure due to ethanol pretreatment. The water content of the aortic wall tissue and the spin-lattice relaxation times (T1) as assessed by proton nuclear magnetic resonance spectroscopy did not change significantly owing to ethanol pretreatment. The optimum condition of 80% ethanol pretreatment almost completely extracted both phospholipids and cholesterol from the aortic wall; despite this, significant calcification occurred. In conclusion, these results clearly demonstrate that ethanol pretreatment is significantly but only partially effective for inhibition of calcification of BPHV aortic wall and this effect may be due in part to lipid extraction and protein structure changes caused by ethanol. It is hypothesized that ethanol pretreatment may be of benefit for preventing bioprosthetic aortic wall calcification only in synergistic combination with another agent. PMID- 9740005 TI - Effect of protein and cell behavior on pattern-grafted thermoresponsive polymer. AB - A thermoresponsive copolymer, poly(Nisopropylacrylamide-co-acrylic acid), was coupled with azidoaniline. The azidophenyl-derivatized copolymer was grafted in a specific pattern on a polystyrene matrix by photolithography. The surface micropattern appeared and disappeared interchangeably, as observed under a phase contrast microscope, by varying the temperature between 10 degrees C and 37 degrees C. The copolymer-grafted polystyrene surface was hydrophobic at 37 degrees C and hydrophilic at 10 degrees C. Albumin and fibronectin adsorption on the matrix was investigated using the fluorescent-labeling method. Fibronectin adsorbed onto both the grafted and nongrafted regions, while albumin adsorbed more onto the nongrafted regions than the grafted regions. Protein adsorption did not affect surface wettability. Mouse fibroblast STO cells were cultured on tissue culture plates pattern-grafted with the thermoresponsive copolymer. Fibronectin adsorption enhanced cell spreading, while albumin reduced it. When the temperature was lowered, the cells selectively detached from the surface areas grafted with the thermoresponsive copolymer when cultured in serum-free medium; the cells partially detached from these areas when cultured in serum containing medium. The effect of serum proteins on cell detachment was similar to that caused by a mixture of albumin and fibronectin. Albumin adsorption did not affect the detachment of cells, while fibronectin adsorption inhibited it. The results of the present study indicate that a pattern-grafted, thermoresponsive, azidophenyl-derivatized copolymer can effectively facilitate selective cell detachment under some conditions such as serum-free culture or preadsorption of albumin. The pattern-grafting technique will be useful for qualitative microscopic comparison of surfaces prepared differently on one chip under the same conditions. PMID- 9740006 TI - Protein encapsulation within polyethylene glycol-coated nanospheres. I. Physicochemical characterization. AB - The development of injectable nanoparticulate "stealth" carriers for protein delivery is a major challenge. We have shown the possibility of entrapping human serum albumin (HSA) in polyethylene glycol (PEG)-coated monodisperse biodegradable nanospheres with a mean diameter of about 200 nm, prepared from amphiphilic diblock PEG-polylactic acid (PLA) copolymers, with loadings up to 9% (w/w). Microscopic techniques and surface analysis studies enabled us to prove that the protein was well entrapped and not adsorbed onto the particle surface. Zeta potential and water uptake studies corroborated that part of the PEG chains are located in the nanosphere matrix. Water uptake in the nanospheres was related to their chemical composition, i.e., the respective wt% of PEG and PLA in the matrix, and not on their fabrication procedure. The hydrophilic PEG blocks absorbed up to 130% (w/w) water, whereas PLA absorbed only about 10% (w/w). However, the rate of swelling at the beginning of the process was related to the structure of the matrix, more particularly to the manner in which PEG was disposed at the surface. Furthermore, it was shown that the PEG "brush" at the nanosphere surface drastically reduces HSA adsorption on the PEG-PLA nanospheres compared to the PLA ones. PMID- 9740007 TI - Reduction of fibrous adhesion formation by a copolymer possessing an affinity for anionic surfaces. AB - Postsurgical adhesions represent a common complication following a variety of surgical procedures. We sought to develop and evaluate a water-soluble polymer that could self-assemble onto tissue surfaces, forming a barrier on the surface. A copolymer was synthesized so as to contain two components: one component adsorbed to the tissue surface, and the other created a steric barrier, thereby preventing cell interactions with the tissue surface, and perhaps altering the wound-healing response that leads to the formation of fibrous adhesions. The component selected for tissue binding was a water-soluble polycation, poly-L lysine, which can bind to negative sites on glycoproteins, proteoglycans, and cells; and the component selected for steric stabilization was polyethylene glycol, a nonionic polymer that interacts poorly with proteins. Efficacy of lavage with an aqueous solution of the copolymer for the prevention of postsurgical abdominopelvic adhesions was assessed following a standard electrocautery injury of the uterine horns of rats. The copolymer resulted in an 88% reduction in the extent of adhesions that formed. In vitro studies designed to investigate the mechanism of this efficacy indicated that the copolymer may both hinder cell-tissue adhesive interactions and alter the process of fibrin formation. PMID- 9740008 TI - Structure-property correlations in a combinatorial library of degradable biomaterials. AB - A combinatorial library of degradable polyarylates was prepared. These polymers are A-B-type copolymers consisting of an alternating sequence of a diphenol and a diacid. The library was prepared by copolymerizing, in all possible combinations, 14 different tyrosine-derived diphenols and eight different aliphatic diacids, resulting in 8 x 14 = 112 distinct polymers. This approach (a) increases the number of available polymeric candidate materials for medical applications, and (b) facilitates the identification of correlations between polymer structure and glass transition temperature, air-water contact angle, mechanical properties, and fibroblast proliferation. The pendent chain and backbone structures were systematically varied by (a) simple homologative variations in the number of methylene groups, (b) substitution of oxygen for methylene groups, and (c) introduction of branched and aromatic structures. The polymers contained within the library exhibited incremental variations in Tg (from 2 degrees C to 91 degrees C) and air-water contact angle (from 64 degrees to 101 degrees ). Fibroblast proliferation (in vitro, serum-containing media) ranged from approximating that measured on tissue culture polystyrene to complete absence of proliferation. Generally, decreased proliferation correlated linearly with increased surface hydrophobicity, except in those polymers derived from oxygen containing diacids in their backbone which were uniformly good growth substrates even if their surfaces were very hydrophobic. In a selected subgroup of polymers, tensile strength of thin solvent cast films ranged from about 6 to 45 MPa, while Young's modulus (stiffness) ranged from about 0.3 to 1.7 GPa. Combinatorial biomaterial libraries such as these tyrosine-derived polyarylates permit the systematic study of material-dependent biological responses and provide the medical device designer with the option to choose a suitable material from a library of related polymers that encompasses a broad range of properties. PMID- 9740009 TI - Effects of dental amalgam and heavy metal cations on cytokine production by peripheral blood mononuclear cells in vitro. AB - The effects of dental amalgam on cytokine production by human peripheral blood mononuclear cells (PBMC) from healthy donors were analyzed. To induce cytokine production, PBMC were stimulated with lipopolysaccharide, phytohemagglutinin, or staphylococcal enterotoxin A and cultured for 48 h in the presence of either freshly prepared amalgam, aged amalgam, or amalgam-conditioned culture medium (ACCM). The concentrations of several cytokines were measured in PBMC supernatants by enzyme-amplified sensitivity immunoassays (EASIAs). Freshly prepared amalgam as well as ACCM induced a decrease in the production of interferon-gamma (IFN-gamma) and interleukin-10 (IL-10), and an increase in the concentrations of tumor necrosis factor-alpha (TNF-alpha). Both fresh amalgam and ACCM showed no effects on IL-2, IL-6, or granulocyte-macrophage colony stimulating factor levels. Amalgam aged for 6 weeks did not affect the concentration of any of the above cytokines. To investigate which heavy metal cations released from amalgam caused the observed immunomodulatory effects, Cu2+, Hg2+, and Sn2+, which were detected in amalgam supernatants by inductively coupled plasma atomic spectrophotometry, were added as salts to the cultures. Cu2+ and Hg2+ induced a decrease in IFN-gamma and IL-10 levels, and Hg2+ an increase in TNF-alpha concentrations. Cytokine production was not significantly modulated by Sn2+. Under these experimental conditions, release of Ag+ into culture medium was not detectable. However, Ag+ markedly suppressed the production of IFN-gamma, IL-10, and TNF-alpha. In summary, our results show that fresh amalgam, but not amalgam aged for 6 weeks, causes changes in the cytokine pattern of PBMC in vitro, and that these effects are due to the release of Cu2+ and Hg2+. PMID- 9740010 TI - Peptide surface modification of poly(tetrafluoroethylene-co-hexafluoropropylene) enhances its interaction with central nervous system neurons. AB - Poly(tetrafluoroethylene-co-hexafluoropropylene) (FEP) film surfaces were chemically surface modified to introduce one of three laminin adhesive peptides: GYIGSR, GRGDS, or SIKVAV. FEP film surfaces were first reduced with sodium naphthalide to introduce surface carbon-carbon double bonds at two reaction conditions: 20 min at -78 degrees C, and 3 h at 25 degrees C. Scanning electron microscopy and atomic force microscopy indicated that surface topography was unaffected by the reaction conditions. Reduced FEP film surfaces were further modified to introduce hydroxyl groups via hydroboration/oxidation or carboxylic acid groups via oxidation. The hydroxyl (FEP-CHxOH) and carboxylic acid (FEP COOH) functionalized surfaces provided reactive handles for peptide coupling using tresyl chloride. Surface elemental composition data, determined from X-ray protoelectron spectroscopy, indicated that equivalent amounts of GYIGSR, GRGDS, and SIKVAV were introduced. Two additional coupling reagents, SMCC and TSU, were compared to tresyl chloride for the coupling of radio-labeled tyrosine of GYIGSR. Between 8 and 150 fmol/cm2 of peptide was introduced to the hydroxyl and carboxylic acid functionalized surfaces, with the tresyl coupling reagent showing the greatest amount of peptide incorporated. The tresyl-coupled peptide-modified surfaces were compared in terms of the response of primary, embryonic hippocampal neurons plated from serum-free medium for 4 days. The number and length of neurites extending from the cell bodies were averaged over 50 cells after 1 and 4 days FEP-CHxO-peptide surfaces had either a greater or equivalent hippocampal neuron interaction than the corresponding FEP-COO-peptide surfaces. All peptide functionalized surfaces had a greater hippocampal neuron interaction than the corresponding FEP-CHxOH, FEP-COOH, and FEP controls after 4 days underlying the importance of the peptides over hydrophilic or hydrophobic surfaces. After 4 days differences in neurite extension were evident among the peptide-functionalized surfaces, with the longest neurites observed on the SIKVAV-functionalized surfaces. PMID- 9740011 TI - Evaluation of proliferation and protein expression of human bone marrow cells cultured on coral crystallized in the aragonite of calcite form. AB - The two crystalline forms of CaCO3, aragonite (from natural coral) and calcite (from natural limestone), have been used with success as bone graft substitutes. However, natural coral transformed into calcite by heating has never been tested. The objective of this work was to study the proliferation and alkaline phosphatase, osteonectin, and osteocalcin expression of human bone marrow cells cultured on CaCO3 crystallized both in the aragonite form (natural coral) and in the calcite form (natural coral modified by heating). The methods used to characterize calcite obtained from the coral were volumic porosimetry, scanning electron microscopy (SEM) and X-ray diffraction. Cell colonization of the material was assessed by SEM performed on days 1, 7, 20, and 30 and [3H]thymidine incorporation was performed on days 3, 7, 12, 18, 25, and 32. Phenotypic expression was assessed by using in situ cytochemistry (alkaline phosphatase), immunocytochemistry (osteonectin and osteocalcin), and hybridization (osteocalcin, beta-actin, and alkaline phosphatase mRNA). Results showed the transformation of aragonite into calcite after heating, the conservation of macroporosity, and a modification of the surface. Calcite appeared to have a smoother and more uniform surface than aragonite crystals. As for [3H]thymidine there was an increase incorporation from days 3 to 18, a stabilization from days 18 to 25, and a decrease from days 25 to 32. After 20 days of culture, immunological studies using monoclonal antibodies to osteocalcin, osteonectin, cytochemical analysis of alkaline phosphatase activity, and in situ hybridization using osteocalcin, beta-actin, and alkaline phosphatase cDNA indicated that the cells had not lost their osteoblastic phenotype. These experiments demonstrate that coral crystallized in the aragonite or calcite form present a similar degree of specific cytocompatibility. PMID- 9740012 TI - Metal wear particle characterization from metal on metal total hip replacements: transmission electron microscopy study of periprosthetic tissues and isolated particles. AB - The less intense tissue reaction around metal on metal total hip replacements (THRs) compared to metal on polyethylene (PE) THRs may be explained by the differences in the characteristics of metal wear particles. In this study, transmission electron microscopy was used to study metal wear particles that were either in situ in cells or had been extracted from the cells by a new technique based on enzymatic tissue digestion. The tissues were obtained from 13 patients undergoing revision of metal on metal THRs with cobalt-chromium-molybdenum (CoCrMo) bearing couples. Most of the CoCrMo wear particles were smaller than 50 nm (range 6-834 nm) and round to oval in shape with irregular boundaries. This size range is considerably smaller than that reported for PE particles. While even a small volume of metal wear will produce high numbers of particles, the apparently less severe local tissue reaction to metal particles may be due to the possibility that corrosion, dissolution, and dissemination of metal particles may result in fewer local biological effects than the long-term retention of PE particles in the periprosthetic tissues. PMID- 9740013 TI - A new drug delivery system with controlled release of antibiotic only in the presence of infection. AB - An ideal drug delivery system (DDS) releases an appropriate drug at specific locations and times. We tried to create a new antibiotic delivery system that releases gentamicin only when wounds are infected by Pseudomonas aeruginosa (P.A.). Exudate from the dorsal pouch of rats infected with P.A. showed significantly higher hydrolytic activity-thrombin-like activity-toward Boc-Val Pro-Arg-MCA than exudate from noninfected wounds. We therefore constructed a device for controlled release of an antimicrobial drug triggered by thrombin-like activity. Briefly, gentamicin was bound to a polyvinyl alcohol derivative (PVA) hydrogel through a newly developed peptide linker cleavable by the proteinase, PVA-(linker)-gentamicin. In vitro experiments showed that proteinases from wounds infected with P.A. cleaved the linker and gentamicin was released while the exudate from noninfected wounds had no hydrolytic activity toward the linker. This device shows potential as an occlusive dressing with an effective antibiotic delivery system for treating infected wounds. PMID- 9740014 TI - Attachment, morphology, and protein expression of rat marrow stromal cells cultured on charged substrate surfaces. AB - Although surface charge has been shown to affect the adhesion and morphology of a variety of cell types, the interactions of bone marrow stromal cells with charged surfaces still remain unclear. A novel electrical stimulation system was used to investigate the interactions between rat bone marrow stromal cells and charged substrates in this study. A conductive and transparent indium tin oxide (ITO) coating was used as an electret substrate. Rat marrow stromal cells were cultured on positive, negative, and uncharged ITO surfaces. Cell attachment, morphology, alkaline phosphatase activity, and expression of osteopontin and collagen type III were assessed using histochemical staining, immunolabeling, and fluorescence microscopy. Voltages of 0.7, 0.8, 0.9, and 1.0 V applied to the substrates created surface potentials but were insufficient to decompose the media. On positively charged ITO, cell attachment was enhanced in serum-supplemented and serum-free media. Furthermore, decreases in cell spreading, alkaline phosphatase activity, and osteopontin were observed in cells grown on the positively charged ITO. These data indicate that positively charged surfaces enhance cell attachment but suppress cell spreading and differentiation of rat marrow stromal cells. PMID- 9740015 TI - In vivo investigation of blood compatibility of titanium oxide films. AB - Titanium oxide films were synthesized by ionbeam-enhanced deposition. The films were prepared by depositing titanium atoms and simultaneously bombarding them with Xe+ ions at an energy of 40 keV in an O2 environment. An in vivo investigation, which entailed implanting low-temperature isotropic pyrolytic carbon (LTI carbon) cylinders, widely used to fabricate artificial heart valves, and titanium-oxide-coated LTI carbon cylinders with diameters of 5 mm and thicknesses of 0.5 mm into the ventral aorta of dogs for 14 days, showed that the amount of thrombus on the titanium-oxide-coated LTI carbon was much less than that formed on the surface of LTI carbon alone. Scanning electron microscopy (SEM) was used to observe the morphology of thrombus. On the titanium oxide films no platelet aggregation was found, almost no red blood cells were damaged, and almost no fibrin was found on the surface. However, all three characteristics were found on the surface of LTI carbon alone, proving that the blood compatibility of titanium oxide films is better than that of LTI carbon and titanium-oxide-coated LTI carbon. PMID- 9740016 TI - Surface characterization and biological properties study of silicone rubber membrane grafted with phospholipid as biomaterial via plasma induced graft copolymerization. AB - Poly(2-methacryloyloxyethyl phosphorylcholine) (pMPC) was grafted onto the surface of a silicon rubber (SR) membrane (pMPC-SR) by plasma induced grafted copolymerization (PIP). Argon plasma was used to activate the SR surfaces. Determination was also made of the influences of grafted copolymerization reaction time, reaction temperature, and monomer concentration on polymerization yield. The surface properties of SR were characterized by ATR-FTIR, ESCA, and SEM. In those analyses the ATR-FTIR spectra indicated that the pMPC grafted onto the SR surface at 1720 and 3300 cm(-1). The elemental composition and different carbon bindings on the surface of the SR were examined by ESCA. An increasing P1s/C1s value g was obtained in the grafted polymerization yield with a concentration of 0.05-0.5M of MPC in the isolated ethanol solution. The surface morphologies of pMPC-SR differed more than those of control and Ar plasma treated surfaces. The difference could have been caused by the homogeneous graft polymerization of pMPC onto the SR membrane. In the biological analyses, protein adsorption on pMPC-SR surfaces was reduced. The reduced level increased with an increase in the pMPC grafted amount. The epithelial cell attachment and growth onto these samples were suppressed. The blood compatibility for a series of pMPC SR surfaces was examined by platelet adhesion. Blood platelet morphologies in contact with the high ratio of pMPC-SR surfaces were maintained, meaning that in this case the release reaction for platelets never occurred. Consequently, the high amount of pMPC-SR surface had excellent blood compatibility, further suggesting that prevention of adhesion, activation of platelets, and adsorption of blood protein could be achieved. PMID- 9740017 TI - Active platelet movements on hydrophobic/hydrophilic microdomain-structured surfaces. AB - The early motion and interaction of platelets on a microdomain-structured block copolymer surface composed of 2-hydroxyethyl methacrylate (HEMA)-styrene were analyzed and compared with those on a compositionally identical random copolymer, homopolymer poly (HEMA) (hydrophilic) and polystyrene (hydrophobic) surfaces. Contacting platelets were quantitatively more active, with motions including rolling, detachment, oscillatory vibration, and change of direction only on the HEMA-St block copolymer surface. Active platelet movements were observed for long time periods (>20 min) on HEMA-St block copolymer surfaces and were distinct from those for inert PSt latex particles on these same surfaces, demonstrating that platelet movements were not due to physical forces such as convection, hydrophobic interactions, or microbrownian movement. To study the cause and mechanism underlying the platelet movements, platelets treated with an adenosine triphosphate (ATP) synthesis inhibition, NaN3, or a membrane skeleton-disrupting chemical agent, dibucaine, were also studied on these surfaces. Both treatments reduced platelet movement and demonstrated that platelets in contact with the HEMA-St block copolymer surface require metabolic processes consuming ATP and involve dynamics of their membrane skeleton. These energy-consuming active movements might explain the previously observed lower platelet activation and low thrombogenicity of the HEMA-St block copolymers. Enhanced platelet movements on the HEMA-St block copolymer surface show that the microdomain surface interacts uniquely with platelets to hinder activation and preserve passive platelet function despite surface contact. PMID- 9740018 TI - Fibrinogen adsorption and host tissue responses to plasma functionalized surfaces. AB - The physical and chemical characteristics of material surfaces are thought to play important roles in biomaterial-mediated tissue responses. To understand the importance of discrete biomaterial chemical characteristics in modifying host tissue responses, we constructed surfaces bearing different functional groups using radio frequency glow discharge plasma polymerization. Surfaces evaluated included those having high concentrations of -OH, -NH2, -CF3, and siloxyl groups. These surfaces and polyethylene terephthalate controls were used to assess the importance of particular physicochemical characteristics in surface:protein:cell interactions both in vitro and in vivo. The results obtained show that surface functionalities do significantly affect both the adsorption and "denaturation" of adsorbed fibrinogen (which is an important mediator of inflammatory responses to biomaterial implants). In addition, these surfaces provoke different degrees of acute inflammatory responses. Interestingly, the amounts of "denatured" fibrinogen that spontaneously accumulate on the individual surfaces correlate closely with the extent of biomaterial-mediated inflammation. These results suggest that surfaces that tend to "irreversibly" bind fibrinogen prompt greater acute inflammatory responses. Unexpectedly, all test surfaces except those bearing a siloxyl group engender relatively similar biomaterial-mediated fibrotic responses. Thus surface functionalities alone may not be sufficient to affect subsequent fibrotic responses. PMID- 9740020 TI - Transcriptional analysis of the Drosophila GABA receptor gene resistance to dieldrin. AB - The Resistance to dieldrin (Rdl) gene encodes a novel subunit of a gamma aminobutyric acid (GABA)-gated chloride ion channel in Drosophila. We were interested in defining the spatial and temporal expression pattern of this gene and in understanding the basis of its regulation. Rdl is expressed in both the embryonic central and peripheral nervous system. Here, we describe the complete Rdl transcription unit (approximately 50 kb) via localization of the flanking transcripts. The Rdl transcript itself is large (8.8 kb) and is composed of a short open reading frame (2 kb) with exceptionally long 5' (1.8-kb) and 3' (5-kb) untranslated regions (UTRs). The correct spatial and temporal expression of Rdl can be rescued by transformation constructs containing only 3.5 kb of DNA, a region which encompasses the transcription start point (tsp). This region also contains sequences strikingly similar to those found in other ion channel gene promoters. Failure of minigene constructs lacking the long 3' UTR to fully rescue both the lethal and resistance phenotypes associated with the Rdl locus might arise owing to the role of these sequences in message stability or trafficking. PMID- 9740019 TI - Insulin-like growth factor-I prevents apoptosis in neurons after nerve growth factor withdrawal. AB - Insulin-like growth factor-I (IGF-I) is emerging as an important growth factor able to modulate the programmed cell death (PCD) pathway mediated by the cysteine dependent aspartate proteases (caspases); however, little is known about the effect of IGF-I after nerve growth factor (NGF) withdrawal in neurons. To begin to understand the neuronal death-sparing effect of IGF-I under NGF-free conditions, we tested whether embryonic sensory dorsal root ganglion neurons (DRG) were able to survive in defined serum-free medium in the presence of IGF-I. We further studied the role of IGF-I signaling and caspase inhibition after NGF withdrawal. NGF withdrawal produced histological changes of apoptosis including chromatin condensation, shrinkage of the perikaryon and nucleus, retention of the plasma membrane, and deletion of single cells. Both IGF-I and Boc-aspartyl (OMe) fluoromethylketone (BAF), a caspase inhibitor, equally reduced apoptosis after NGF withdrawal. The antiapoptotic effect of IGF-I was completely blocked by LY294002, an inhibitor of PI 3-kinase signaling, but not by the mitogen-activated protein (MAP) kinase/extracellular signal-regulated protein kinase (ERK) activated protein kinase inhibitor PD98059. Functional IGF-I receptors were extensively expressed both in rat and human DRG neurons, although they were most abundant in the neuronal growth cone. Collectively, these findings indicate that IGF-I, signaling though the PI-3 kinase pathway, is important in modulating PCD in cultured DRG neurons after NGF withdrawal, and IGF-I may be important in DRG embryogenesis. PMID- 9740021 TI - neuroD induces photoreceptor cell overproduction in vivo and de novo generation in vitro. AB - The molecular mechanisms underlying the generation of the various types of cells in the vertebrate retina are largely unknown. We investigated the possibility that genes belonging to the basic helix-loop-helix (bHLH) family of transcriptional factors participate in cell-type specification during retinal neurogenesis. Chick neuroD was isolated from an embryonic cDNA library and its deduced amino acid sequence showed 75% identity with mouse neuroD. In situ hybridization showed that neuroD was expressed in cells located at the outer portion of the developing retinal neuroepithelium, the location where prospective photoreceptors reside. Misexpression of neuroD in retinal neuroepithelium through replication-competent, transformation-deficient retroviruses produced a retina with three, instead of two, layers of photoreceptor cells; the number of cells that express visinin, a marker for cone photoreceptors, increased over 50% compared to control embryos misexpressing the green fluorescent protein. No significant changes were observed in the number of other retinal neurons, including those that express RA4 (ganglion cells), pax6 (ganglion cells and amacrine cells), and chx10 (bipolar cells). Retroviral-driven misexpression of neuroD in monolayer cultures of retinal pigment epithelium yielded de novo production of photoreceptor cells with no other types of retinal neurons detected. We propose that neuroD is important for photoreceptor cell production in the vertebrate retina. PMID- 9740022 TI - Muscle degeneration following remote nerve injury. AB - Muscle depends upon innervation and contraction to maintain a differentiated state. Denervation can therefore induce muscle atrophy. In grasshoppers, muscle degeneration can also be triggered by the severing of a leg during autotomy. In this case, the muscles that degenerate are neither damaged nor denervated. This phenomenon suggests the existence of transneuronal mechanisms that influence muscle survival. To characterize this autotomy-induced process, we studied the degeneration of a thoracic tergotrochanteral depressor muscle (M#133b,c) subsequent to the shedding of a hindlimb in the grasshoppers Barytettix psolus and Barytettix humphreysii. Both histochemical and electrophysiological methods were used to follow muscle degeneration 1, 3, 5, 10, and 15 days postautotomy. Muscle fibers began to show denervation-like electrophysiological changes (i.e., depolarized resting membrane potentials and postinhibitory rebound) as soon as 3 days postautotomy. By 10 days, significant muscle degeneration was evident and electrophysiological changes were found in all animals tested. Muscle anatomical degeneration was not induced by synaptic transmission failure, because neuromuscular transmission was maintained in most fibers. The rate of muscle degeneration was not constant. Between 1 and 10 days, mean fiber cross-sectional area did not change on the autotomized side, although this is normally a time of muscle growth. However after 10 days, cross-sectional area became drastically reduced and the number of muscle fibers within M#133b,c was decreased. The variability in rate of fiber degeneration was not dependent upon fiber type, since M#133b,c only contains fast-type fibers. PMID- 9740023 TI - Analysis of neuronal and glial phenotypes in brains of mice deficient in leukemia inhibitory factor. AB - Leukemia inhibitory factor (LIF) can regulate the survival and differentiation of certain neurons and glial cells in culture. To determine the role of this cytokine in the central nervous system in vivo, we examined the brains of young and adult mice in which the LIF gene was disrupted. Immunohistochemical staining of neurons for choline acetyltransferase, tyrosine hydroxylase, serotonin, parvalbumin, calbindin, neuropeptide Y, vasoactive intestinal polypeptide, and calcitonin gene-related peptide revealed no significant differences between null mutant and wild-type (WT) brains. In contrast, analysis of glial phenotypes demonstrated striking deficits in the LIF-knockout brain. Staining with several anti-glial fibrillary acidic protein (GFAP) antibodies showed that the number of GFAP-positive cells in various regions of the hippocampus in the female mutant is much lower than in the WT. The null male hippocampus also displays a significant, though less marked deficit. The number of astrocytes in the mutant hippocampus, as determined by S-100 staining, is not, however, significantly different from WT. In addition, quantification of immunohistochemical staining of female, but not male, mutants reveals a significant deficit in myelin basic protein content in three brain regions, suggesting alterations in oligodendrocytes as well. Thus, while overall brain histology appears normal, the absence of LIF in vivo leads to specific, sexually dimorphic alterations in glial phenotype. PMID- 9740024 TI - Molecular cloning of a lobster Gbeta subunit and Gbeta expression in olfactory receptor neuron dendrites and brain neuropil. AB - We have isolated from the olfactory organ of the American lobster (Homarus americanus) two cDNA clones with homology to beta subunits of G proteins. LobGbeta1 contained a complete open reading frame that predicted an amino acid sequence with >80% identity to Gbeta sequences from other species. LobGbeta2 was a fragment of an open reading frame whose predicted amino acid sequence had 65 69% identity to other Gbeta sequences. LobGbeta2 mRNA was not detectable in the brain, eye plus eyestalk, leg, dactyl, olfactory organ, or tail muscle. In contrast, lobGbeta1 was expressed in all these tissues as a single mRNA species of 6.4 kb and a protein of 37 kD. In the brain and olfactory organ, Gbeta immunoreactivity was almost exclusively confined to neurites: the neuropil regions of the brain and the outer dendrites of the olfactory receptor neurons. Coimmunoprecipitation revealed that lobster Gbeta interacted with both Galpha s and Galpha q. LobGbeta1 is likely to be involved in a wide range of signaling events including olfactory transduction and synaptic transmission in the brain. PMID- 9740026 TI - Photoperiod and testosterone independently affect vocal control region volumes in adolescent male songbirds. AB - Previously, we found that, unlike adults, adolescent male dark-eyed juncos (Junco hyemalis) maintained large Area X volumes despite having low plasma testosterone concentrations. Other studies indicate that photoperiod may act independently of testosterone to modulate vocal control region (VCR) volumes in adult songbirds. In the present study, we investigated the effects of testosterone and photoperiod on the volumes of four VCRs in adolescent male juncos. To test the hypothesis that VCR volumes in these males are testosterone independent, we treated birds exposed to short days with testosterone and later compared their VCR volumes with those of birds exposed to short days without testosterone. To examine whether photoperiod alone could affect VCR volumes independent of testosterone, we measured these volumes in photorefractory birds exposed to long photoperiod without testosterone. Administering testosterone induced singing, yet increased the volume of only one VCR, the robust nucleus of the anterior neostriatum (RA). In contrast, long photoperiod increased several VCR volumes (Area X, higher vocal center, and RA) despite low testosterone levels, but did not induce singing. Our results suggest a limited role for testosterone, but an important role for photoperiod, in controlling VCR volumes in adolescent male juncos. In addition, the results demonstrate that singing behavior can be induced in adolescent males without a concomitant increase in most VCR volumes. PMID- 9740025 TI - Physiological stress and nerve growth factor treatment regulate stress-activated protein kinase activity in PC12 cells. AB - The stress-activated protein kinases (SAPKs) are differentially activated by a variety of cellular stressors in PC12 cells. SAPK activation has been linked to the induction of apoptotic cell death upon serum withdrawal from undifferentiated cells or following nerve growth factor (NGF) withdrawal of neuronally differentiated PC12 cells. However, withdrawal of trophic support from differentiated cells led to only a very modest elevation of SAPK activity and led us to investigate the basis of the relative insensitivity of these enzymes to stressors. NGF-stimulated differentiation of the cells resulted in the elevation of basal SAPK activity to levels four- to sevenfold greater than in untreated cells, which was correlated with an approximate fivefold increase in SAPK protein levels. Paradoxically, in NGF-differentiated PC12 cells, exposure to cellular stressors provoked a proportionately smaller stimulation of SAPK activity than that observed in naive cells, despite the presence of much higher levels of SAPK protein. The insensitivity of SAPK to activation by stressors was reflective of the activity of the SAPK activator SEK, whose activation was also diminished following NGF differentiation of the cells. The data demonstrate that SAPKs are subject to complex controls through both induction of SAPK expression and the regulation mediated by upstream elements within this pathway. PMID- 9740027 TI - Regulation of a Purkinje cell-specific promoter by homeodomain proteins: repression by engrailed-2 vs. synergistic activation by Hoxa5 and Hoxb7. AB - We have previously demonstrated that a short sequence element (L7ATE) within the proximal promoter of a Purkinje cell-specific gene, pcp-2(L7), is required for the normal pattern of expression of the gene in the cerebellum of transgenic mice. The presence of a series of TAAT sequence motifs in this element suggested its interaction with homeodomain proteins. To extend these observations, degenerate oligonucleotides were used to clone by reverse-transcriptase polymerase chain reaction members of the mouse Hox gene family expressed in neonatal cerebellum but not forebrain. Two of these, HoxB7 and HoxA5, are continuously expressed from the neonatal period into adult stages in cerebellar Purkinje cells. These Hox proteins are shown to synergistically activate the L7 promoter by cotransfection assay in vitro. In contrast, another homeodomain protein that is normally expressed in Purkinje cells only during the embryonic period, En-2, has a negative effect on L7 gene expression. These data suggest a biphasic, combinatorial control mechanism for the Purkinje cell-specific expression of the pcp-2(L7) gene. PMID- 9740029 TI - 1998 Basmajian/Williams & Wilkins Award: Judith E. Anderson, University of Manitoba. PMID- 9740028 TI - Placodal origin of Brn-3-expressing cranial sensory neurons. AB - The Brn-3 class of POU-domain transcription factors includes three genes in mammals which have key roles in the development of specific groups of sensory neurons. Here, we have identified three avian genes which correspond to the murine genes Brn-3.0, Brn-3.1, and Brn-3.2. Using an in situ hybridization probe generic for this gene class, the earliest detectable expression of Brn-3 in the chick is at stage 15, in placodal and migrating precursors of the trigeminal ganglion. By stage 19, Brn-3.0 protein is detectable in the trigeminal and vestibulocochlear ganglia with Brn-3.0-specific antisera, and Brn-3 message expression has extended to the dorsal root ganglia. At later stages, when condensation of the trigeminal ganglion is complete, Brn-3.0-immunoreactive neurons are concentrated in the portion of the ganglion distal to the brain stem. To examine the developmental origin of the Brn-3 expressing cells, we combined lipophilic dye (DiI) labeling with in situ hybridization. DiI labeling of the placodal surface ectoderm and of premigratory neural crest cells in the neural tube reveals that all, or nearly all, of the Brn3-expressing neurons in the trigeminal ganglia are derived from the sensory placodes and not from the neural crest, and thus, that Brn-3 is an early marker of the placode-derived sensory neural lineage. PMID- 9740030 TI - The Charles Judson Herrick Award: Georg F. Striedter, University of California, Irvine. PMID- 9740031 TI - 1998 R.R. Bensley Award: Keith E. Mostov, University of California at San Francisco. PMID- 9740032 TI - The 1998 Henry Gray Award. PMID- 9740034 TI - Morphology, paleoanthropology, and Neanderthals. AB - Morphology carries the primary signal of events in the evolutionary history of any group of organisms but has been relatively neglected by paleoanthropologists, those who study the history of the human species. Partly this is the result of historical influences, but it is also due to a rather fundamentalist adherence among paleoanthropologists to the tenets of the Neodarwinian Evolutionary Synthesis. The result has been a general paleoanthropological desire to project the species Homo sapiens back into the past as far and in as linear a manner as possible. However, it is clear that the human fossil record, like that of most other taxa, reveals a consistent pattern of systematic diversity--a diversity totally unreflected in the conventional minimalist interpretation of that record. Thus, the Neanderthals, both morphologically and behaviorally as distinctive a group of hominids as ever existed, are conventionally classified simply as a subspecies of our own species Homo sapiens--a classification that robs these extinct relatives of their evolutionary individuality. Only when we recognize the Neanderthals as a historically distinctive evolutionary entity, demanding understanding in its own terms, will we be able to do them proper justice. And we will only be able to do this by restoring morphology to its proper place of primacy in human evolutionary studies. PMID- 9740033 TI - Progress in the study of brain evolution: from speculative theories to testable hypotheses. AB - Darwin's theory of evolution raised the question of how the human brain differs from that of other animals and how it is the same. Early students of brain evolution had constructed rather grand but speculative theories which stated that brains evolved in a linear manner, from fish to man and from simple to complex. These speculations were soundly refuted, however, as contemporary comparative neurobiologists used powerful new techniques and methodologies to discover that complex brains have evolved several times independently among vertebrates (e.g., within teleost fishes and birds) and that brain complexity has actually decreased in the lineages leading to modern salamanders and lungfishes. Moreover, the old idea that brains evolved by the sequential addition of new components has now been replaced by the working hypothesis that brains generally evolve by the divergent modification of preexisting parts. Speculative theories have thus been replaced by testable hypotheses, and current efforts in the field are aimed at making phylogenetic hypotheses even more testable. Particularly promising new directions for comparative neurobiology include (1) the integration of comparative neuroanatomy with comparative embryology and developmental genetics in order to test phylogenetic hypotheses at a mechanistic level, (2) research into how evolutionary changes in the structure of neural circuits are related to evolutionary changes in circuit function and animal behavior, and (3) the analysis of independently evolved similarities to discover general rules about how brains may or may not change during the course of evolution. PMID- 9740035 TI - Current concepts of climbing fiber function. AB - This review examines several of the current postulates regarding the function of one of the most intriguing afferent systems in the brain, the climbing fiber system. The fact that these afferents are activated under a variety of conditions has contributed substantially to the diversity of postulates that have been proposed. In part because of the unique anatomical relationship between individual climbing fibers and the dendritic tree of Purkinje cells, these afferents have been proposed as a key input in establishing long-term plastic changes in the cerebellar cortex. This concept is contrasted with other postulates proposing that the heterosynaptic action of this system produces a short-lasting enhancement rather than a long-term depression of Purkinje cell responsiveness. Although a generally accepted view regarding climbing fiber function does not exist, this review emphasizes the extensive functional insights that have been reported and supports the notion that progress toward a complete understanding of these afferents will require an integration of their morphological characteristics with the fundamental physiological properties of their responses assessed in a variety of contexts and conditions. PMID- 9740037 TI - Bladder and rectum volume estimations using CT and stereology. AB - Twelve prostate cancer patients underwent a treatment planning CT prior to radiotherapy. Stereologic method based on Cavalieri principle was applied to CT images. Systematic and random sampling of CT slices were performed with alternative ways for assessing the volumes of bladder and rectum. The contribution of point counting to the precision of the obtained volume estimations was analysed. It was found that 100-150 test points counted on only 5 7 systematically sampled slices through bladder or rectum suffice for reliable volume estimations of the above organs. Also, the superiority of systematic vs random sampling was confirmed. The suggested volumetric method gives the possibility of generating unbiased and efficient bladder and rectum volume assessments directly from CT data with great saving in labour. PMID- 9740036 TI - Endocardial border identification in two-dimensional echocardiographic images: review of methods. AB - A basic goal in the analysis of echocardiographic images is to identify the locations of the endocardial borders. This is necessary in order to create three dimensional reconstructions, and to derive precise quantitative parameters from the images. Endocardial borders can be identified manually; however, this is time consuming, inconvenient, and liable to human subjectivity. For this reason there has been a great deal of interest in automatic border identification methods. In recent years numerous studies have been published, describing a wide variety of proposed border identification algorithms. In spite of the large number of published studies, however, the relative merits of the various methods remain unclear. In this paper the various analysis techniques are reviewed and discussed. The reasons for the lack of definitive conclusions regarding the utility of these methods are described. Finally, the basic methodological requirements necessary for empirical evaluation of border identification techniques are described. PMID- 9740038 TI - Segmentation of the hippocampus from brain MRI using deformable contours. AB - The application of a discrete dynamic contour model for segmentation of the hippocampus from brain MRI has been investigated. Solutions to several common problems of dynamic contours in this case and similar cases have been developed. A new method for extracting the discontinuous boundary of a structure with multiple edges near the structure has been developed. The method is based on detecting and following edges by external forces. The reliability of the final contour and the model stability have been improved by using a continuous mapping of the external energy and limiting movements of the contour. The problem of optimizing the internal force weight has been overcome by making it dependent on the amount of the external force. Finally, the results of applying the proposed algorithm, which implements the above modifications, to multiple applications have been evaluated. PMID- 9740039 TI - Quantification of the local heartwall motion by magnetic resonance myocardial tagging. AB - Sophisticated magnetic resonance tagging techniques provide powerful tools for the non-invasive assessment of the local heartwall motion towards a deeper fundamental understanding of local heart function. For the extraction of motion data from the time series of magnetic resonance tagged images and for the visualization of the local heartwall motion a new image analysis procedure has been developed. New parameters have been derived which allows quantification of the motion patterns and are highly sensitive to any changes in these patterns. The new procedure has been applied for heart motion analysis in healthy volunteers and in patient collectives with different heart diseases. The achieved results are summarized and discussed. PMID- 9740040 TI - Three-dimensional modeling of biopsy protocols for localized prostate cancer. AB - Prostate cancer is the most common malignant tumor in American men, yet only a small percentage of men will develop clinically significant disease. Needle core biopsies are used to confirm the presence of cancer prior to surgery. While needle core biopsies have shown some ability to predict tumor volume and grade in prostatectomy specimens, for the individual patient they are neither sensitive nor specific enough to guide therapy. In this paper, we describe a system for simulating needle biopsies on three-dimensional models of cancerous prostates reconstructed from serial sections. First we segment the serial sections, delineating tumors and landmarks. Next, we register the sections using a color merging scheme, and reconstruct the three-dimensional model using modified-shape based interpolation. The resulting volume can be rendered, and simulated needle core biopsies can be taken from the reconstructed model. We use our system to simulate two different biopsy protocols on a reconstructed prostate specimen. PMID- 9740041 TI - The value of dynamic MR imaging for hypointensity lesions of the peripheral zone of the prostate. AB - The aim was to evaluate the role of dynamic magnetic resonance (MR) imaging for prostatic carcinoma. Forty-two men with clinical suspicion of a prostatic carcinoma underwent MR imaging. Dynamic MR was performed, followed by postcontrast T1-weighted imaging with fat suppression. Histologic diagnosis was 21 prostatic carcinomas (in 19 patients), 21 benign tissues, and 2 chronic prostatitis. The diagnostic accuracy was 75% for T2-weighted images, and 79% for dynamic images. The accuracy of the combination of dynamic MR images with postcontrast T1-weighted images was 82%. It was concluded that dynamic MR imaging was useful in differentiation of low intensity lesions in the peripheral zone. PMID- 9740042 TI - Development of phantoms for spiral CT. AB - PURPOSE: This paper reports on the development of a new phantom for spiral CT. The phantom meets the increased demands on phantom z-axis uniformity in order that objects from the CT slice, immediately above and below the CT slice of interest, do not contribute perturbing information to the reconstructed CT slice. MATERIAL AND METHODS: The phantom depends on formulation of tissue-like materials that can be cast and produced in both geometric and anthropomorphic shapes with sufficient z-axis length to enable unperturbed CT slices of test objects of interest. These materials are then used to produce a series of test objects of CT image quality including low contrast samples that do not require volume averaging or mixing of solutions, and that can reflect sub-slice thickness test objects and supra-slice thickness test objects. RESULTS: The overall phantom and its individual test objects provides meaningful tests of spiral CT image quality including slice sensitivity, CT number linearity and tests of high and low contrast resolution. Schematic designs and actual CT scans are shown. CONCLUSION: The new spiral phantom appears to meet the increased demands of spiral CT on phantom design, particularly z-axis length, and requirements for low contrast resolution test objects. PMID- 9740043 TI - A case of abducens neurinoma mimicking acoustic neurinoma. AB - Neurinoma arising from the abducens nerve independent of neurofibromatosis has been rarely reported in the existing literature. We present a case of abducens neurinoma which was confirmed during a surgical excision. A 31-year-old female had experienced a hearing disturbance for the past 8 months. Abduction of the right eye ball was almost full. Magnetic resonance images showed a tumor having a size of 44 x 33 X 33 mm at the right cerebellopontine angle. Neuro-otological examination revealed the findings specific to acoustic neurinoma. Surgical excision confirmed that the tumor originated from the abducens nerve. The tumor located at the cavernous sinus or cerebellopontine angle might originate from the abducens nerve, though the abduction of eye ball is not in any way impaired. PMID- 9740044 TI - Choroid plexus papilloma of the temporal horn associated with transtentorial herniation. AB - In the pediatric age group choroid plexus papillomas are most commonly located in the trigones and bodies of the lateral ventricles. This paper reports a 1-year old patient with choroid plexus papilloma of the temporal horn of the lateral ventricle associated with transtentorial herniation. PMID- 9740045 TI - Proteome and proteomics: new technologies, new concepts, and new words. AB - The goal of proteomics is a comprehensive, quantitative description of protein expression and its changes under the influence of biological perturbations such as disease or drug treatment. Quantitative analysis of protein expression data obtained by high-throughput methods has led us to define the concept of "regulatory homology" and use it to begin to elucidate the basic structure of gene expression control in vivo. Such investigations lay the groundwork for construction of comprehensive databases of mechanisms (cataloguing possible biological outcomes), the next logical step after the soon to be completed cataloguing of genes and gene products. Mechanism databases provide a roadmap towards effective therapeutic intervention that is more direct than that offered by conventional genomics approaches. PMID- 9740046 TI - Proteome analysis: biological assay or data archive? AB - In this review we examine the current state of proteome analysis. There are three main issues discussed: why it is necessary to study proteomes; how proteomes can be analyzed with current technology; and how proteome analysis can be used to enhance biological research. We conclude that proteome analysis is an essential tool in the understanding of regulated biological systems. Current technology, while still mostly limited to the more abundant proteins, enables the use of proteome analysis both to establish databases of proteins present, and to perform biological assays involving measurement of multiple variables. We believe that the utility of proteome analysis in future biological research will continue to be enhanced by further improvements in analytical technology. PMID- 9740047 TI - Glycobiology and proteomics: is mass spectrometry the Holy Grail? AB - One characteristic of glycoproteins is that they are separated by two-dimensional electrophoresis (2-D PAGE) into typical 'trains' of protein spots which separate on the basis of different isoelectric point (pI) and/or molecular mass. The pattern of these trains often varies in development and disease. While the isoforms differ both in the number of sites of glycosylation and the types of carbohydrate attached to the protein, classical methods of glycan analysis are insensitive at the levels typically separated by 2-D PAGE. Developments in mass spectrometry technologies have enabled the characterization of most of the oligosaccharide attributes to be determined on picomole amounts of protein. These techniques are beginning to allow the glycoform heterogeneity on 2-D separated glycoproteins to be analyzed. PMID- 9740048 TI - The Australian Proteome Analysis Facility (APAF): assembling large scale proteomics through integration and automation. AB - The field of proteomics opens new possibilities for the mass screening of proteins from many different sources. While genomics is well understood to be a big science field, proteomics is just emerging as such. This paper describes the setting up of the first national proteomics facility. The facility has been funded by the Australian government and this funding has allowed the design of purpose built, integrated laboratories with state of the art equipment for large scale proteome research. PMID- 9740049 TI - Proteomic analysis of enamel cells from developing rat teeth: big returns from a small tissue. AB - Poor understanding of the molecular links between disturbed calcium regulation in cells and disease remains a problem of considerable biomedical importance. Dental enamel cells might provide useful insights to this problem since they handle calcium in bulk without suffering its cytotoxic effects. Two practical challenges hindered investigation of calcium handling mechanisms in enamel cells--paucity of molecular information and limited availability of sample from developing rat teeth, the experimental system of choice. This paper outlines the microscale proteomic approaches we applied to overcome these difficulties and reviews several major findings that ensued from initial characterization of the enamel cell proteome. Enamel cells are now established as a powerful model for fundamental calcium research and have provided outcomes of broad biological relevance, including the discovery of a new endoplasmic reticulum protein. Future proteomic approaches that might benefit understanding of function are discussed. PMID- 9740050 TI - New zwitterionic detergents improve the analysis of membrane proteins by two dimensional electrophoresis. AB - Severe quantitative loss of protein is often observed in high-resolution two dimensional electrophoresis of membrane proteins, while the resolution is usually not affected. To improve the solubility of proteins in this technique, we tested denaturing cocktails containing various detergents and chaotropes. Best results were obtained with a denaturing solution containing urea, thiourea, and zwitterionic detergents, synthesized for this purpose. Among the dozen detergents synthesized and tested, amidosulfobetaines with an alkyl tail containing 14-16 carbons proved most efficient, solubilizing previously undetected membrane proteins. PMID- 9740051 TI - Suppression effects in enzymatic peptide ladder sequencing using ultraviolet - matrix assisted laser desorption/ionization - mass spectormetry. AB - The techniques of enzymatic and chemical peptide ladder sequencing, coupled with ultraviolet - matrix assisted laser desorption/ionization - mass spectrometry (UV MALDI-MS) have been improving continuously in the last five years and have now become important tools for primary structure identification. In this work, signal suppression effects, appearing in UV-MALDI-MS (excitation 337 nm) of ladder peptides, were investigated using the 17-amino acid peptide dynorphin A. We show, with examples of simple "two-peptide" systems and more complex "multi-peptide" systems, that suppression effects do in fact exist. The magnitude of the observed suppression is strongly dependent upon both the nature and the amount of the suppressing peptide. Suppression behavior of individual ladder peptides was investigated on equimolar mixtures of ten ladder peptides. Significant signal suppression was recorded for all ladder peptides, with some of them being approximately 170 times lower in signal intensity than the pure, i.e., unsuppressed peptide at the same concentration. For the investigated system- dynorphin A, 4-hydroxy-alpha-cyanocinnamic acid (4-HCCA) matrix, UV excitation--a correlation between the extent of suppression and an intractable combination of peptide hydrophobicity and the presence of several basic amino acids can be seen. PMID- 9740053 TI - An algorithm for the identification of proteins using peptides with ragged N- or C-termini generated by sequential endo- and exopeptidase digestions. AB - We have developed an algorithm (MassDynSearch) for identifying proteins using a combination of peptide masses with small associated sequences (tags). Unlike the approach developed by Matthias Mann, 'Tag searching', in which the sequence tags are generated by gas phase fragmentation of peptides in a mass spectrometer, 'Rag Tag' searching uses peptide tags which are generated enzymatically or chemically. The protein is digested either chemically or with an endopeptidase and the resultant mixture is then subjected to partial exopeptidase degradation. The mixture is analyzed by matrix assisted laser desorption and ionization time of flight mass spectrometry and a list of intact peptide masses is generated, each associated with a set of degradation product masses which serve as unique tags. These 'tagged masses' are used as the input to an algorithm we have written, MassDynSearch, which searches protein and DNA databases for proteins which contain similar tagged motifs. The method is simple, rapid and can be fully automated. The main advantage of this approach is that the specificity of the initial digestion is unimportant since multiple peptides with tags are used to search the database. This is especially useful for proteins like membrane, cytoskeletal, and other proteins where specific endopeptidases are less efficient and lower specificity proteases such as chymotrypsin, pepsin, and elastase must be used. PMID- 9740052 TI - Identification of yeast proteins from two-dimensional gels: working out spot cross-contamination. AB - With the complete sequence of the yeast genome now available, efforts by many laboratories are underway to identify each of the spots on two-dimensional (2-D) gels corresponding to the most abundant yeast proteins. The high mass accuracy now attainable using matrix assisted laser desorption/ionization (MALDI)-mass spectrometry equipped with delayed extraction simplifies the process of identification, such that many spots can be unambiguously identified in a short period of time merely by using peptide mass fingerprinting and generally available database matching programs. Although it is not always possible to match spots between gels run by different laboratories, proteins generally yield the same abundant proteolytic fragments when tryptic digestions are performed. Databases containing these signature peptides not only simplify the task of reidentifying proteins from different gels, but also make it possible to identify small amounts of cross-contaminating proteins from different spots, as well as common extraneous contaminants such as human keratins. In this paper, we present data on the identification of > 20 previously unreported yeast proteins from 2-D gels. Some novel proteins were identified from randomly analyzed spots. Focusing on 14 spots in a narrow-pH-range gel, we demonstrate how organizing peak-table data and peptide match-list data into databases enables the identification of a larger percentage of the peaks. PMID- 9740054 TI - Towards an automated approach for protein identification in proteome projects. AB - The development of automated, high throughput technologies for the rapid identification of proteins is essential for large-scale proteome projects. While a degree of automation already exists in some stages of the protein identification process, such as automated acquisition of matrix assisted laser desorption ionisation-time of flight (MALDI-TOF) mass spectra, efficient interfaces between different stages are still lacking. We report the development of a highly automated, integrated system for large scale identification of proteins separated by two-dimensional gel electrophoresis (2-DE), based on peptide mass fingerprinting. A prototype robotic system was used to image and excise 288 protein spots from an amido black stained polyvinylidene difluoride (PVDF) blot. Protein samples were enzymatically digested with a commercial automated liquid handling system. MALDI-TOF mass spectrometry was used to acquire mass spectra automatically, and the data analysed with novel automated peptide mass fingerprinting database interrogation software. Using this highly automated system, we were able to identify 95 proteins on the basis of peptide mass fingerprinting, isoelectric point and molecular weight, in a period of less than ten working days. Advantages, problems, and future developments in robotic excision systems, liquid handling, and automated database interrogation software are discussed. PMID- 9740055 TI - Structural determination of N-linked carbohydrates by matrix-assisted laser desorption/ionization-mass spectrometry following enzymatic release within sodium dodecyl sulphate-polyacrylamide electrophoresis gels: application to species specific glycosylation of alpha1-acid glycoprotein. AB - This paper describes a sensitive method for analysis of N-linked carbohydrates released enzymatically from within the gel following separation of glycoproteins (50-100 pmols) by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS PAGE). The separated bands containing the glycoproteins were cut from the gel, destained, reduced and alkylated. N-linked glycans were then released by in-gel incubation with peptide N-glycosidase-F (PNGase-F) and extracted with water and acetonitrile. Sialic acid-containing glycans were converted into methyl esters by reaction with methyl iodide, salts and reagents were removed by passage through a mixed-bed column of ion-exchange resins and the glycans were examined by matrix assisted laser desorption/ionization (MALDI)-mass spectrometry. Structural determination of the released glycans was performed by exoglycosidase digestion. Following glycan release and extraction, the protein could be digested within the gel with trypsin, and the masses of the tryptic peptides could be compared with those generated from a sequence database for protein identification. The method is applied to the analysis of N-linked glycans from alpha1-acid glycoprotein from man, cow, sheep and dog. Major species-specific differences in glycosylation were found. Thus, although all four species used N-acetyl-neuraminic acid, only cow and sheep additionally used N-glycolyl-neuraminic acid. Biantennary glycans were the predominant carbohydrates in cow, sheep and dog but man produced more triantennary glycans and a substantial amount of tetraantennary sugars. Fucosylation was only found in glycans from man and cow and both cow and sheep glycans were found to have beta1-3- and well as beta1-4-linked galactose residues in the antennae. PMID- 9740056 TI - '98 Escherichia coli SWISS-2DPAGE database update. AB - The combination of two-dimensional polyacrylamide gel electrophoresis (2-D PAGE), computer image analysis and several protein identification techniques allowed the Escherichia coli SWISS-2DPAGE database to be established. This is part of the ExPASy molecular biology server accessible through the WWW at the URL address http://www.expasy.ch/ch2d/ch2d-top.html . Here we report recent progress in the development of the E. coli SWISS-2DPAGE database. Proteins were separated with immobilized pH gradients in the first dimension and sodium dodecyl sulfate polyacrylamide gel electrophoresis in the second dimension. To increase the resolution of the separation and thus the number of identified proteins, a variety of wide and narrow range immobilized pH gradients were used in the first dimension. Micropreparative gels were electroblotted onto polyvinylidene difluoride membranes and spots were visualized by amido black staining. Protein identification techniques such as amino acid composition analysis, gel comparison and microsequencing were used, as well as a recently described Edman "sequence tag" approach. Some of the above identification techniques were coupled with database searching tools. Currently 231 polypeptides are identified on the E. coli SWISS-2DPAGE map: 64 have been identified by N-terminal microsequencing, 39 by amino acid composition, and 82 by sequence tag. Of 153 proteins putatively identified by gel comparison, 65 have been confirmed. Many proteins have been identified using more than one technique. Faster progress in the E. coli proteome project will now be possible with advances in biochemical methodology and with the completion of the entire E. coli genome. PMID- 9740057 TI - Determination of plasmid-encoded functions in Rhizobium leguminosarum biovar trifolii using proteome analysis of plasmid-cured derivatives. AB - We have used proteome analysis of derivatives of R. leguminosarum biovar trifolii strain ANU843, cured of indigenous plasmids by a direct selection system, to investigate plasmid-encoded functions. Under the conditions used, the plasmid encoded gene products contributed to only a small proportion of the 2000 proteins visualised in the two-dimensional (2-D) protein map of strain ANU843. The level of synthesis of thirty-nine proteins was affected after curing of either plasmid a, c or e. The differences observed upon plasmid curing included: protein loss, up/down-regulation of specific proteins and novel synthesis of some proteins. This suggests that a complex interplay between the cured plasmid and the remaining replicons is occurring. Twenty-two proteins appeared to be absent in the cured strains and these presumably are encoded by plasmid genes. Of these, a small heat shock protein, a cold shock protein, a hypothetical YTFG-29.7 kDa protein, and the alpha and beta subunits of the electron transfer flavoprotein were identified by N-terminal microsequencing and predicted to be encoded by plasmid e. Four of the sequenced proteins putatively encoded on plasmid e and two encoded on plasmid c were novel. In addition, curing of plasmid e and c consistently decreased the levels of 3-isopropylmalate dehydratase and malate dehydrogenase, respectively, suggesting that levels of these proteins may be influenced by plasmid-encoded functions. A protein with homology to 4 oxalocrotonate tautomerase, which is involved in the biodegradation of phenolic compounds, was found to be newly synthesised in the strain cured of plasmid e. Proteome analysis provides a sensitive tool to examine the functional organisation of the Rhizobium genome and the global gene interactions which occur between the different replicons. PMID- 9740058 TI - Strategies towards a better understanding of antibiotic action: folate pathway inhibition in Haemophilus influenzae as an example. AB - Two-dimensional electrophoresis was applied to the global analysis of the cellular response of Haemophilus influenzae to sulfamethoxazole and trimethoprim, both inhibitors of tetrahydrofolate synthesis. Deregulation of the synthesis rate of 118 proteins, involved in different metabolic pathways, was observed. The regulation of the genes involved in the metabolism of the amino acids methionine, threonine, serine, glycine, and aspartate was investigated in detail by analysis of protein synthesis and Northern hybridization. The results suggested that the synthesis of methionine biosynthetic enzymes in H. influenzae is regulated in a similar fashion as in Escherichia coli. A good correlation between the results obtained by Northern hybridization and quantification of protein synthesis was observed. In contrast to trimethoprim, sulfamethoxazole triggered the increased synthesis of the heat shock proteins DnaK, GroEL, and GroES. PMID- 9740059 TI - A novel method for quantitative analysis of recombinant secreted proteins using two-dimensional electrophoresis. AB - We describe a two-dimensional polyacrylamide gel electrophoresis (2-D PAGE)-based approach for detecting and quantifying secreted recombinant proteins in media conditioned by baby hamster kidney (BHK) cells. Seven secreted proteins were analyzed in this system: leptin, thrombopoietin, thrombin, glycoprotein 130, soluble interleukin-2 receptor, and two novel sequences obtained from sequence database searches. BHK cells transfected with plasmids encoding each of these proteins, and cells transfected with empty plasmids (control cells), were metabolically labeled and the resulting conditioned media were analyzed by 2-D PAGE. Gel images derived from cells expressing recombinant proteins were compared with images from control cells in order to identify spots corresponding to the expressed proteins. All seven of the test proteins were successfully detected using this method. The sensitivity of the system was evaluated by diluting samples derived from high-expressing clones with conditioned media from control cells. The sensitivity of detection was protein-dependent, but recombinant proteins expressed at levels as low as 10 ng/mL could be detected. Quantification of recombinant protein levels was achieved by measuring spot intensities using phosphorimager analysis. The intensity of spots corresponding to recombinant proteins were compared with the spot intensity of an endogenous BHK protein which had been calibrated to a known standard. Estimates of the levels of expressed protein determined using this technique correlated with the levels determined using standard affinity assays. We conclude that this system provides a reliable method for quantifying levels of protein expression when specific assays are unavailable. PMID- 9740060 TI - New insights into cyclosporine A nephrotoxicity by proteome analysis. AB - Using two-dimensional gel electrophoresis (2-DE), we recently discovered an association between decreased calcium-binding protein, calbindin-D 28 kDa, urinary calcium wasting and intratubular corticomedullary calcifications in rat kidney. This observation prompted us to investigate kidney tissues of other species, including man. In this paper we show that in dogs and monkeys, which are generally devoid of cyclosporine A (CsA)-mediated nephrotoxicity, renal calbindin levels were not affected by the CsA treatment whereas in CsA-treated human kidney transplant recipients with renal vascular or tubular toxicity, a marked decrease in renal calbindin-D 28 kDa protein level was found in most of the kidney biopsy sections. The present results strongly suggest that calbindin is a marker for CsA nephrotoxicity. The discovery of calbindin-D 28 kDa being involved in CsA toxicity has evolved from the application of 2-DE and has not been reported previously, proving that proteomics can provide essential information in mechanistic toxicology. Considering the current improvements in proteome methods it is expected that high throughput proteomics will become an indispensable tool in preclinical safety testing. PMID- 9740061 TI - Mining the human proteome: experience with the human lymphoid protein database. AB - We have undertaken an effort in the past five years aimed at developing a database of lymphoid proteins detectable by two-dimensional (2-D) polyacrylamide gel electrophoresis. The database contains 2-D patterns and derived information pertaining to: (i) polypeptide constituents of unstimulated and stimulated mature T cells and immature thymocytes; (ii) cultured T cells and cell lines that have been manipulated by transfection with a variety of constructs or by treatment with specific agents; (iii) single cell-derived T and B cell clones; (iv) cells obtained from patients with lymphoproliferative disorders and leukemia; and (v) a variety of other relevant cell populations. The database has experienced a substantial expansion in 2-D patterns it contains, numbering currently 9167 individual 2-D patterns. This number represents a fraction of the 30,682 2-D patterns maintained in our databases. The capacity to design and undertake experiments, produce high-quality 2-D patterns, and to undertake simple or rudimentary analyses of 2-D patterns to meet the basic needs of the experiments for which the 2-D gels were produced has exceeded the capacity to fully and uniformly integrate information generated from any gel image or experiment, across all images and experiments. While only a fraction of the information in the 2-D patterns contained in the lymphoid database has been mined, novel findings derived from querying the database point to the merits of this protein based approach. Additional resources have recently been put into place to mine more effectively data pertaining to protein expression in lymphoid cells. PMID- 9740062 TI - Charge heterogeneity of macrophage migration inhibitory factor (MIF) in human liver and breast tissue. AB - Macrophage migration inhibitory factor (MIF) is an ubiquitous protein playing various immunological and hormonal roles. Theoretical electrophoretic coordinates calculated from protein sequence in the SWISS-PROT database (AC P14174) are 12 kDa and pI 8.24. Using two-dimensional (2-D) immunoblotting, we have detected isoelectric forms at ca. 11.9 kDa, with pI values of 7.8 and 6.98 in human liver tissue, breast tissue and a cell line and in preparations of human MIF expressed in E. coli. This evidence suggests that MIF charge heterogeneity originates from a post-translational modification not requiring eukaryote-specific enzymes. We have also detected in human liver a minor immunoreactive spot at pI 6.23, which coincides with the MIF spot in the liver map in SWISS-2DPAGE. The pI 6.23 isoform also conceivably derives from post-translational modification, as MIF is known to be encoded in the human genome by a single copy gene. PMID- 9740063 TI - Proteomic changes associated with degeneration of myelin-forming cells in the central nervous system of c-myc transgenic mice. AB - Myelin is necessary for the conduction of high frequency and high velocity nerve impulses in the central nervous system of mammals, and severe neurological disturbances develop as a result of myelin loss. In this report, we have characterized changes in the brain proteomic profile of transgenic mice that develop a c-myc-induced degenerative disorder of myelin. Marked differences were seen in the accumulation of cytoskeletal proteins associated with the pathological condition fibrous gliosis in the optic nerves of affected animals, including upregulation of glial fibrillary acid protein and vimentin. In addition, the expression of several major myelin proteins, including five isoforms of myelin basic protein, four isoforms of cyclic nucleotide 3' phosphodiesterase, and myelin-associated glycoprotein, was markedly reduced in the brains of c-myc transgenic mice as revealed by immunocytochemistry and by two dimensional immunoblots. A number of novel proteomic disease marker candidates were revealed, which displayed pronounced changes in their expression profile. Sequence determination of these proteins and molecular cloning of their mRNAs will provide an opportunity to further evaluate their roles in the disease process. PMID- 9740064 TI - Protein changes observed in pacing-induced heart failure using two-dimensional electrophoresis. AB - Rapid ventricular pacing in dogs results in a low output cardiomyopathic state which is similar to idiopathic dilated cardiomyopathy in man. However, the pathophysiological mechanisms which cause this failure following pacing are unknown. Five dogs underwent rapid ventricular pacing. Hearts were stimulated at 245 beats per min (bpm) for four weeks and then reduced to 190 bpm to stabilize the failure. Six unoperated dogs were used as controls. This paper compares the two-dimensional gel electrophoresis (2-DE) protein patterns of left ventricular samples from the paced myocardium with the control dogs. Changes in protein expression were analyzed qualitatively and semi-quantitatively. In the paced dog samples 69 protein spots were significantly altered of which 42 were decreased and 27 were elevated. One qualitative change was observed: elongation factor Tu was present only the control hearts. Of these proteins, 20 have been identified by a combination of N-terminal protein microsequencing, peptide mass profiling by mass spectrometry, amino acid compositional analysis, and by comparison with databases of canine and human ventricular proteins. Ten of these are associated with mitochondria and energy production, including: pyruvate dehydrogenase E1 component, isocitrate dehydrogenase subunit alpha, HSP60 and HSP70, creatine kinase M and fatty acid binding protein. The cytoskeletal protein desmin was detected in reduced quantities and a spot corresponding to a fragment of desmin was increased. These results indicate that the development of heart failure in the paced dog involves alterations in mitochondrial energy production, the cytoskeleton and calcium activation. PMID- 9740065 TI - Cardiac protein abnormalities in dilated cardiomyopathy detected by two dimensional polyacrylamide gel electrophoresis. AB - The aim of the investigation was to determine whether there are specific global quantitative and qualitative changes in protein expression in heart tissue from patients with dilated cardiomyopathy (DCM) compared with ischaemic heart disease and undiseased tissue. Two-dimensional (2-D) polyacrylamide gel electrophoresis and computer analysis was used to study protein alteration in DCM biopsy material (n=28) compared with donor heart biopsy samples (n=9) and explanted hearts from individuals suffering from ischaemic heart disease (IHD; n = 21). A total of 88 proteins displayed decreased abundance in DCM versus IHD material while five proteins had elevated levels in the DCM group (p<0.01). The most prominent changes occurred in the contractile protein myosin light chain 2 and in a group of proteins identified as desmin. These changes do not appear to be artefactual degradation events occurring during sample processing. These proteins are not apparent in electrophoretic separations of vascular tissue or cultured endothelial cells, mesothelial cells or cardiac fibroblasts, which are clearly distinguishable from the 2-D protein patterns of whole heart and of isolated cardiac myocytes and do not appear to reflect variations in the cellular composition of biopsy samples. The different protein patterns observed in cardiomyopathy showed no obvious relationship with New York Heart Association (NYHA) functional class or haemodynamic parameters. The study has demonstrated significant alterations in quantitative protein expression in the DCM heart which would have serious implications for myocyte function. These changes might be explained by altered protease activity in DCM which could exacerbate contractile dysfunction in the failing heart. PMID- 9740066 TI - Effects of renovascular hypertension on myocardial protein patterns: analysis by computer-assisted two-dimensional gel electrophoresis. AB - Hypertensive heart disease caused by renovascular hypertension reflects the response of the heart to an increased afterload and neurohormonal stimulation. We hypothesized that in this condition the composition of the myocardial proteins of rats was altered. To identify yet unknown quantitative and qualitative differences in myocardial proteins in renovascular hypertensive heart disease, we analyzed protein patterns by computer-assisted two-dimensional polyacrylamide large gel electrophoresis. Renovascular hypertension was induced by placing a silver clip on the left renal artery in 9-week-old rat siblings. Sham-operated animals served as controls. Systolic blood pressure (197 +/- 19 mm Hg) and heart/body weight ratios (0.36 +/- 0.04%) were significantly increased in the hypertensive animals. Twenty protein patterns from the left ventricle of five hypertensive and five control rats were compared. The molecular mass and isoelectric point (pI) of proteins spots ranging from 13 to 100 kDa and from 4.5 to 8.5, respectively, were determined using marker proteins. In total, 761 +/- 88 protein spots were resolved in all twenty gels. For the quantitative data analysis a univariate (Mann-Whitney test) as well as a multivariate statistical approach (correspondence analysis) were applied. Only one myocardial protein spot (molecular mass = 41.3 kDa; pI = 6.3) was decreased by more than twofold (p < 0.05) in renovascular hypertension. The vast majority of spots did not indicate a significant alteration of intensity. Left ventricular hypertrophy in early renovascular hypertension induces a form of myocardial hypertrophy that conserves the naturally occurring protein expression pattern. PMID- 9740067 TI - The concept of Tenju-gann, or "natural-end cancer". PMID- 9740068 TI - Cancer death and older age. PMID- 9740070 TI - Expression of CD44 variant proteins in adenocarcinoma of Barrett's esophagus and its relation to prognosis. AB - BACKGROUND: None of the commonly used staging criteria accurately determine the prognosis of a patient with adenocarcinoma of Barrett's esophagus. The authors therefore assessed the expression pattern and prognostic impact of CD44 standard and CD44 isoforms CD44v4, v5,v6,v7, and v10 in adenocarcinoma of Barrett's esophagus. METHODS: Specimens from 41 patients with adenocarcinoma of Barrett's esophagus who underwent esophageal resection were embedded in paraffin and studied immunohistochemically to determine the expression of CD44 splice variants. Histomorphologic parameters and survival time were not known at the time of the investigation. RESULTS: Correlations between favorable clinical or histomorphologic parameters and CD44s or any of the split variants could not be established. Down-regulation of CD44s and the split variant v10 was significantly correlated with pT classification. Furthermore, down-regulation of CD44v10 and up regulation of CD44v7 were significantly correlated with ploidy. There was a significant correlation between CD44s and split variants in tumorous and nontumorous tissue from the same patient. Down-regulation of CD44s and CD44v4 had a significant influence on prognosis in that it was associated with shortened life expectancy. Multivariate analysis revealed that the expression of CD44v4 was an independent factor in prognosis. CONCLUSIONS: The results obtained for this small patient sample suggest that CD44v4 is a new independent prognostic parameter for adenocarcinoma of Barrett's esophagus that can be determined preoperatively by biopsy. It may therefore be helpful in planning therapy by allowing the identification of patients who may benefit from esophageal resection as well as those who are at high risk for morbidity and mortality even when the tumor is otherwise resectable. Further studies of larger patient samples are required to validate the results of the current study. PMID- 9740069 TI - Nasopharyngeal carcinoma in situ: two cases of an emerging diagnostic entity. AB - BACKGROUND: The association of Epstein-Barr virus (EBV) with the oncogenesis of nasopharyngeal carcinoma (NPC) is well established. Latent infection by EBV with clonal proliferation has also been demonstrated in preinvasive lesions of NPC. In situ hybridization for EBV-encoded RNA (ISH EBER) now serves as an ancillary test in the definitive diagnosis of these lesions. METHODS: Two cases of nasopharyngeal carcinoma in situ (NPCIS) are presented in this study. Their biopsies were studied by ordinary light microscopy, the ISH EBER technique, and immunostaining for bcl-2. Tissue samples from 100 high risk subjects negative for NPC and NPCIS, who served as controls, were also studied using the ISH EBER technique. RESULTS: NPCIS was characterized by abnormal light microscopic appearance as well as positive staining by the ISH EBER technique; these features were not observed in samples from the 100 high risk subjects. Immunostaining for bcl-2 protein was positive but less specific. Postradiotherapy biopsies of the two patients were negative for NPCIS. CONCLUSIONS: With the help of the ISH EBER technique, the diagnosis of NPCIS is now possible in routine surgical pathology. As this entity is rare, it is necessary to have a high degree of suspicion when evaluating biopsies from high risk individuals. Radiotherapy for patients with NPCIS is justified in view of the risk of cancer progression and the possibility of a coexisting invasive carcinoma. PMID- 9740071 TI - S-phase fraction can predict event free survival in patients with pT2-T3N0M0 colorectal carcinoma: implications for adjuvant chemotherapy. AB - BACKGROUND: Adjuvant chemotherapy for colorectal carcinoma was found to improve survival of patients with American Joint Committee on Cancer/International Union Against Cancer Stage III disease. The usefulness of chemotherapy in patients with Stage II disease continues to be debated, and it is likely that only those patients with a poor prognosis should receive adjuvant chemotherapy. Biologic prognostic factors may allow further insight into the optimal treatment strategy for patients with Stage II or earlier disease. In this study the prognostic role of S-phase fraction (SPF) determined by flow cytometry was assessed in patients with Stage I-II colorectal carcinoma. METHODS: Specimens of surgically resected colorectal carcinoma were examined for SPF by flow cytometric DNA analysis. Consecutive patients referred to the study institution were considered eligible for this study. The main inclusion criteria were a Stage I-II tumor together with sufficient tumor material and adequate follow-up information. For each stage of disease, SPF data were associated with the recurrence rate and the disease free survival (DFS). RESULTS: Analysis was performed on 167 patients (65 with Stage I disease and 102 with Stage II disease). Among Stage I patients, SPF was high in 20 patients and low in 45 patients. In Stage II patients, there were 36 patients with low SPF and 66 patients with high SPF. In both stages, the recurrence rate and DFS were significantly worse for the subgroups of patients with high SPF. CONCLUSIONS: SPF has revealed prognostic differences among patients with surgically resected Stage I-II colorectal carcinoma. These data should be considered for planning future trials in the adjuvant setting because patients with high SPF may benefit from adjuvant chemotherapy. PMID- 9740072 TI - Resection of both hepatic and pulmonary metastases in patients with colorectal carcinoma. AB - BACKGROUND: More than 40% of patients who undergo curative resection of advanced colorectal carcinoma can be expected to have recurrence of the disease. The most frequent sites of recurrence are the liver (33% of patients) and lung (22%). Interest has therefore focused on treating hepatic or pulmonary metastases, or both, to improve the outcomes of these patients. Although surgical resection has become an increasingly accepted treatment for resectable localized hepatic or localized pulmonary metastases from colorectal carcinoma, the value of aggressive surgery for the removal of both hepatic and pulmonary metastases from patients with primary colorectal carcinoma remains to be clarified. METHODS: Data on 30 patients who had undergone resection of both hepatic and pulmonary metastases from colorectal carcinoma were included in the study. RESULTS: Independent, significant prognostic features were found to be the time that hepatic or pulmonary metastases occurred and the distribution of pulmonary metastases. Median survival times were 30 months (range, 7-108 months) after resection of both hepatic and pulmonary metastases and 48.5 months (range, 11-149 months) after excision of the primary colorectal tumor. Actuarial 1-, 3-, and 5-year survival after resection of both hepatic and pulmonary metastases was 86.7%, 49.3%, and 43.8%, respectively. No perioperative mortality occurred. There were three cases of minor morbidity, which the authors considered acceptable. CONCLUSIONS: Resection of both hepatic and pulmonary metastases from colorectal carcinoma may help to prolong the survival of a small group of patients with these metastases. PMID- 9740073 TI - Serum concentration of CD44 variant 6 and its relation to prognosis in patients with gastric carcinoma. AB - BACKGROUND: The expression of variant isoforms of CD44 is correlated with the ability of tumor cells to metastasize in some clinical carcinomas. In this study, the serum concentration of soluble splice isoforms of CD44 that shared exon variant 6 (sCD44v6) were measured and the histologic expression of CD44v6 in tumors from patients with gastric carcinoma examined. METHODS: Serum samples were obtained from 102 patients with primary gastric carcinoma before surgery and serum levels of sCD44v6 were determined with an enzyme-linked immunoadsorbent assay. The expression of CD44v6 in tumors was examined by immunohistochemical staining. RESULTS: Both the serum concentration of sCD44v6 and its expression in tumors were associated significantly with the depth of invasion of the tumor, lymph node metastasis, and clinical stage in patients with diffuse type gastric carcinoma. Among patients with gastric carcinoma, the serum level of sCD44v6 was higher in those with CD44v6 positive tumor cells than in those with CD44v6 negative tumor cells. The serum level of sCD44v6 was a prognostic indicator in patients with diffuse type gastric carcinoma, as was the histologic expression of CD44v6. However, neither CD44v6 nor sCD44v6 was a predictor of survival time in patients with intestinal type gastric carcinoma. CONCLUSIONS: It appears that CD44v6 and sCD44v6 are related to the progression of diffuse type gastric carcinoma. An elevated serum level of sCD44v6 may be useful as a prognostic indicator in patients with diffuse type gastric carcinoma. PMID- 9740074 TI - A phase II study of all-trans-retinoic acid plus cisplatin and etoposide in patients with extensive stage small cell lung carcinoma: an Eastern Cooperative Oncology Group Study. AB - BACKGROUND: The dysregulation of both myc gene expression and retinoid signaling pathways commonly occurs in small cell lung carcinoma (SCLC). Because preclinical data showed that all-trans-retinoic acid (RA) inhibited SCLC growth, altered myc expression, and blocked transition to a treatment-resistant phenotype, a Phase II trial was designed to determine the effects of the combination of RA, cisplatin, and etoposide in patients with SCLC. METHODS: Patients with untreated, extensive stage SCLC were treated with up to 8 cycles of cisplatin, 60 mg/m2, intravenously (i.v.) on Day 1 and etoposide, 120 mg/m2, i.v. on Days 1-3 in addition to up to 1 year of oral RA, 150 mg/m2/day. RESULTS: Of 22 assessable patients 1 had a complete response and 9 had a partial response, for an overall response rate of 45% (95% confidence interval, 24-68%). The median survival was 10.9 months and the 1-year survival was 41%. The median duration of chemotherapy was 6 cycles and the median duration of RA treatment was 2.8 months. Thirteen patients discontinued RA prematurely due to toxicity and only 4 responders were receiving RA at the time of recurrence. Toxicity-limiting RA treatment mainly was comprised of mucocutaneous changes and headaches. CONCLUSIONS: RA at a dose of 150 mg/m2/day was tolerated poorly in combination with cisplatin plus etoposide, leading to early discontinuation of RA in the majority of patients. The hematologic toxicity, response rate, and survival were similar to those associated with cisplatin and etoposide in prior trials. Further studies with more active and less toxic agents will be required to determine the role of retinoids in the treatment of SCLC. PMID- 9740075 TI - Molecular alterations to human chromosome 3p loci in neuroendocrine lung tumors. AB - BACKGROUND: The origins of and interrelations between low grade and high grade neuroendocrine lung tumors, typical and atypical carcinoids, and small cell lung carcinoma (SCLC) have not been elucidated. Karyotypic and molecular genetic studies have demonstrated deletions in 3p in 100% of SCLCs and the candidate lung tumor suppressor gene, FHIT, at 3p14.2 is not expressed in the majority of SCLCs. Similar studies of typical and atypical carcinoids could clarify the interrelations among these tumors. METHODS: For molecular genetic analyses, archival carcinoids and paired normal cells were microdissected from paraffin sections, deparaffinized, and DNA prepared. Oligonucleotide primer pairs for 12 microsatellite markers mapping between 3p14.2 and 3p21.3 were used to amplify allelic DNA fragments from 13 typical and 6 atypical carcinoids. In addition, an independent series of archival sections of carcinoids and SCLCs was tested by immunohistochemistry for expression of Fhit protein. RESULTS: Of the six atypical carcinoids examined, three had lost an allele at all informative markers, whereas one had lost alleles in two distinct regions and two showed allele loss in a subregion of the chromosome region tested. Of the 13 typical carcinoids, 3 showed allele loss at only 1 or 2 loci each. Typical carcinoids, similar to normal lung epithelia, were strongly positive for the cytoplasmic Fhit protein, SCLCs were uniformly negative, and atypical carcinoids appeared to express an intermediate level of Fhit protein. CONCLUSIONS: Loss of heterozygosity at 3p14.2-p21.3 is significantly more extensive in all atypical carcinoids. Atypical carcinoids, which exhibit clinicopathologic features intermediate between typical carcinoids and small cell carcinomas and have been considered well differentiated neuroendocrine carcinomas, also are intermediate between typical carcinoids and SCLC on the basis of extent of loss of 3p alleles and reduced expression of Fhit protein. PMID- 9740076 TI - A parallel study of in vitro sensitivity to benzo[a]pyrene diol epoxide and bleomycin in lung carcinoma cases and controls. AB - BACKGROUND: Because only a fraction of smokers develop neoplastic lesions, host factors may affect their susceptibility to the carcinogenic effects of tobacco smoke. Benzo[a]pyrene diol epoxide (BPDE) is the metabolic product of benzo[a]pyrene (B[a]P), a constituent of tobacco smoke. Therefore, BPDE sensitivity may shed some light on smoking-related carcinogenesis. METHODS: First, differential BPDE sensitivity was tested in five lymphoblastoid cell lines. Then sensitivity to BPDE and bleomycin (an excellent lung carcinoma risk predictor) was tested in parallel in the lymphocytes of 57 lung carcinoma cases and 82 controls. RESULTS: The optimal BPDE treatment duration was 24 hours. The xeroderma pigmentosum cell line was the most sensitive, followed by head and neck cancer, ataxia telangiectasia, and normal cells. The mean breaks per cell for cases and controls were 0.78 and 0.46, respectively (P < 0.0001). BPDE sensitivity was significantly associated with lung carcinoma, with an odds ratio (OR) of 7.26, compared with an OR of 4.56 for bleomycin sensitivity. There was also a dose-response correlation between the quartiles of BPDE-induced breaks and lung carcinoma risk, with ORs of 2.39, 3.12, and 15.03. It is noteworthy that individuals who were sensitive to both BPDE and bleomycin had a significantly increased OR of 38.36. CONCLUSIONS: BPDE sensitivity may be a biologic marker to identify individuals who are susceptible to the carcinogenic effects of tobacco smoke. BPDE and bleomycin sensitivity might represent different repair or sensitivity pathways; however, when these assays are used in parallel, they might refine our ability to identify high risk individuals. PMID- 9740077 TI - Desmoplastic and desmoplastic neurotropic melanoma: experience with 280 patients. AB - BACKGROUND: It has been suggested that desmoplastic melanoma (DM) and desmoplastic neurotropic melanoma (DNM) are associated with worse prognoses and higher local recurrence rates than other forms of melanoma. In the current study, a large series of patients with DM and DNM treated at a tertiary referral center was reviewed. METHODS: For 190 patients with DM and 90 patients with DNM accrued over a 10-year period, clinical features were recorded and all available histopathology was reviewed. The associations between clinical and pathologic variables, biologic behavior, and eventual outcome were analyzed. RESULTS: The male-to-female ratio was 1.75:1 and the median patient age 61 years. The median tumor thickness was 2.5 mm, and 44% of cases were amelanotic. Five-year survival was 75%. Significant predictors of overall survival were a high mitotic rate (P=0.003) and tumor thickness (P=0.011). All the DNMs exceeded 1.5 mm in thickness and were graded as Clark's level IV or V. There was a significant increase in local recurrence when neurotropism was present (P < 0.001). The rate of local recurrence was not higher for DM than for other cutaneous melanomas. CONCLUSIONS: There was no statistically significant difference in survival for patients with DM and those with DNM, and overall survival for both was similar to that for patients with other cutaneous melanomas. However, there was a lower rate of regional lymph node metastasis at initial presentation and as the first recurrence for both DM and DNM. The local recurrence rate was higher when the surgical clearance margin was <1 cm and when neurotropism was present. PMID- 9740078 TI - Oral doxifluridine plus levoleucovorin in elderly patients with advanced breast cancer. AB - BACKGROUND: There currently is no agreement regarding the appropriate treatment of elderly patients with advanced breast carcinoma (ABC). Doxifluridine (5-dFUR), a prodrug of 5-fluorouracil, has been found to be effective in this entity, but its use is limited by neurotoxicity and cardiotoxicity that are not observed when the oral formulation is used. The objective of this Phase II trial was to evaluate the effectiveness and tolerability of oral 5-dFUR, biomodulated with levoleucovorin (1-leucovorin), in elderly patients (age > 70 years) with ABC. METHODS: 5-dFUR was administered orally at 600 mg/m2 twice daily for 4 consecutive days every 12 days, and oral 1-leucovorin was administered as 25 mg 2 hours before each 5-dFUR administration. Response was assessed every five cycles according to the World Health Organization criteria. In the presence of response or stable disease, the patients were treated for a maximum of 15 cycles. RESULTS: Seventy-three eligible patients were enrolled, 27 of whom had been pretreated with chemotherapy and/or hormonotherapy; all were assessable for response and toxicity after a median follow-up of 15 months. The objective response rate was 26% (95% confidence interval, 17.4-45.4). Regression predominantly occurred in the presence of soft tissue involvement (skin, lymph nodes, and breast). The median time to response was 2 months (range, 1-2 months) and the median response duration was 7 months (range, 2-17+ months). The median survival was 24 months (range, 2-42+ months). The treatment was very well tolerated, and the side effects were manageable and always reversible. CONCLUSIONS: The results of the current study show that 5-dFUR plus 1-leucovorin, both given orally, are associated with excellent patient compliance. Although the results are suboptimal in terms of an objective response, this characteristic could allow 5-dFUR to be used in elderly patients considered unsuitable for "aggressive" chemotherapy. PMID- 9740080 TI - Matrix metalloproteinase-2 immunoreactive protein: a marker of aggressiveness in breast carcinoma. AB - BACKGROUND: Previous studies have shown that matrix metalloproteinase-2 (MMP-2) (a 72-kilodalton Type IV collagenase/gelatinase A) is associated with breast carcinoma, but to the authors' knowledge there are no reports showing that it is prognostic for overall survival. METHODS: Expression of the immunoreactive protein for MMP-2 was evaluated in tissue sections from primary breast carcinomas of 177 patients with a monoclonal antibody to MMP-2 using an immunohistochemical technique. RESULTS: Approximately 84% of the samples were MMP-2 positive, with 22% being strongly positive. Positive MMP-2 immunostaining was prognostic for shortened survival. After 10 years 56% of the patients with tumors that were strongly positive for MMP-2 were alive, whereas 88% of patients with an MMP-2 negative tumor and 70% of patients with weakly or moderately positive tumors were still alive (chi-square test = 7.4; P < 0.01, log rank analysis). MMP-2 positivity was linked with an unfavorable prognosis regardless of the age of the patient, tumor grade, receptor status of the tumor, and stage of disease. These results were confirmed by a multivariate analysis in which MMP-2 positivity emerged as an independent prognostic factor for poor survival. CONCLUSIONS: To the authors' knowledge this study is the first time that MMP-2 immunoreactive protein has been associated strongly with a shortened survival independent of major prognostic indicators in patients with primary breast carcinoma, increasing the risk of death 3.6-fold during the first 10 years of follow-up. PMID- 9740079 TI - Anastrozole versus megestrol acetate in the treatment of postmenopausal women with advanced breast carcinoma: results of a survival update based on a combined analysis of data from two mature phase III trials. Arimidex Study Group. AB - BACKGROUND: This report presents the results of a survival update based on the combined data from two studies that compared the efficacy and tolerability of anastrozole (1 or 10 mg once daily), a selective, nonsteroidal aromatase inhibitor administered orally, and megestrol acetate (40 mg 4 times daily) in the treatment of postmenopausal women with advanced breast carcinoma whose disease had progressed after treatment with tamoxifen. METHODS: Two randomized, parallel group, multicenter trials were conducted, involving a total of 764 patients. The two trials were identical in design; both were double blind for anastrozole and open label for megestrol acetate. Overview analyses were conducted with the intent of strengthening the interpretation of results from each trial. The median follow-up duration for this survival update was 31 months. RESULTS: At the clinical dose of 1 mg daily, anastrozole demonstrated a statistically significant survival advantage over megestrol acetate, with a hazard ratio of 0.78 (P < 0.025)(0.60 < 97.5% confidence interval [CI] <1.0). The 1 mg anastrozole group also had a longer median time to death (26.7 months) compared with 22.5 months for the megestrol acetate group. The 10 mg anastrozole group also had a survival benefit over the megestrol acetate group, with a hazard ratio of 0.83 (P=0.09, not significant)(0.64 < 97.5% CI < 1.1). Higher 2-year survival rates were observed for both anastrozole treatment groups than for the megestrol acetate group (56.1%, 54.6%, and 46.3% for the groups given 1 mg anastrozole, 10 mg anastrozole, and megestrol acetate, respectively). CONCLUSIONS: This combined analysis of two trials of postmenopausal patients with advanced breast carcinoma has clearly demonstrated that, after disease progression with tamoxifen, treatment with anastrozole 1 mg once daily results in a statistically and clinically significant advantage over a standard treatment, megestrol acetate. This important benefit, in addition to the good tolerability profile of anastrozole, supports the use of this drug as a valuable new treatment option for this patient population. PMID- 9740081 TI - Adenocarcinoma arising in extragonadal endometriosis: an immunohistochemical study. AB - BACKGROUND: Malignant transformation is an infrequent but reported complication of endometriosis. Previous reports of these cases have been limited to clinicopathologic studies based on routine histologic examination of these tumors, whereas, to the authors' knowledge, characterization of these lesions based on immunophenotype and hormone receptor and oncoprotein expression has not been described. METHODS: Using commercially available monoclonal antibodies, the authors studied three recent cases of adenocarcinoma arising in extragonadal endometriosis using paraffin immunohistochemistry. Proteins examined included different cytokeratin (CK) subtypes, as well as hormone receptor status, proliferation rate, and oncoprotein expression. RESULTS: All three cases presented clinically and macroscopically as colonic masses, and the tumors expressed an endometrial CK phenotype (CK7+, CK20-). In contrast, the adjacent benign colonic epithelium expressed the expected opposite phenotype (CK7-, CK20+). Estrogen receptor (ER) and progesterone receptor (PR) were expressed in one of the three tumors. Interestingly, in the ER/PR negative tumors, receptor expression was present in areas of benign endometriosis adjacent to malignancy, suggesting a loss of receptor expression with malignant transformation. The tumors also were examined for proliferation by Ki-67, and the expression of oncoproteins c-erb B-2, p53, cyclin D1, and bcl-2. All cases of malignancy had a high proliferation rate as measured by Ki-67, which was in contrast to areas of benign endometriosis which had a low proliferation rate. Of the other oncoproteins only p53 protein was detected at a significant level in all three cases. Cyclin D1 was overexpressed in two of the three cases. c-erb B2 and bcl-2 overexpression was not detected. CONCLUSIONS: The results of the current study 1) show the utility of CK subtypes in confirming endometrioid phenotype in tumors arising in extragonadal endometriosis with colonic involvement and 2) suggest that loss of hormone receptor expression and p53 oncoprotein abnormalities may be involved as mechanisms in malignant transformation in extragonadal endometriosis. PMID- 9740082 TI - A worse prognosis for men with testicular atrophy at therapeutic orchiectomy for prostate carcinoma. AB - BACKGROUND: The significance of testicular atrophy at the time of therapeutic orchiectomy for prostate carcinoma has not been examined even though pretreatment hypogonadism has been associated with poor prognosis during chemical androgen ablation for these tumors. METHODS: Survival after therapeutic orchiectomy was determined for 78 men with prostate carcinoma and related to the histologic severity of testicular atrophy. Included in analysis were the presence or absence of prior radiation therapy, tumor grade and stage at diagnosis, host age, obesity, and smoking habits. RESULTS: Among 35 men who underwent therapeutic orchiectomy for progressive disease after primary radiation therapy to the prostate bed, the 25 men with testicular atrophy had worse 5-year, tumor specific, postorchiectomy survival than the 10 men without testicular atrophy (30% vs. 89%) (P=0.02). These 25 men had tumors of more advanced stage and greater undifferentiation at the time of diagnosis an average of 45 months before orchiectomy, but neither characteristic was related to postorchiectomy survival. Among 25 men with Stage D2 disease (American Urologic Association staging system) with orchiectomy as the primary treatment, the 7 men with testicular atrophy more often had undifferentiated tumors and had lower 2-year tumor specific survival than the 18 men without atrophy (43% vs. 72% ) (P > 0.10). CONCLUSIONS: Testicular atrophy at the time of therapeutic orchiectomy for prostate carcinoma is associated with poor postorchiectomy prognosis in men with prior prostate bed radiation therapy and perhaps in men without prior radiation. The association may reflect a high frequency of inherently more aggressive tumors (often relatively nonandrogen-dependent) among those tumors that are progressing in hypogonadal men. PMID- 9740083 TI - Testicular atrophy in therapeutic orchiectomy specimens from men with prostate carcinoma: association with prior prostate bed radiation and older age. AB - BACKGROUND: The significance of testicular atrophy at the time of therapeutic orchiectomy for prostate carcinoma has not been examined even though hypogonadism may occur after prostate bed radiation therapy for these tumors, may itself be symptomatic, and also may be associated with poor tumor prognosis. METHODS: Therapeutic orchiectomy specimens from 78 men with prostate carcinoma and no preceding hormonal therapy were evaluated histologically for atrophy. Observations were related to prior radiation therapy, tumor grade and stage diagnosis, host age, obesity, and smoking habits. RESULTS: Thirty-five men who previously received radiation therapy to the prostate bed had testicular atrophy more frequently than 43 men without prior radiation (71% vs. 28%) (P < 0.001). In men without prior radiation, atrophy was less common in specimens from those age < 70 years than in specimens from men age > 70 years (7% vs. 38%) (P < 0.04). In men with prior radiation, prominent atrophy occurred with similar frequency in specimens from both younger and older men, and was more frequent in specimens obtained within 3 years after radiation therapy than in specimens obtained after longer postradiation intervals (89% vs. 53%) (P < 0.001). CONCLUSIONS: Testicular atrophy at the time of therapeutic orchiectomy for men with prostate carcinoma is much more common in patients with prior prostate bed radiation therapy. Available evidence suggests that this association may reflect both radiation-induced testicular injury and more frequent early tumor recurrence in men with atrophy preceding their radiation therapy. PMID- 9740084 TI - Cause of death in men diagnosed with prostate carcinoma. AB - BACKGROUND: Prostate carcinoma is one of the leading causes of death in men. Although the mortality rate is high, it still may underestimate the number of deaths associated with the disease. This study was conducted to compare causes of death among men previously diagnosed with prostate carcinoma and to examine the extent to which differences in cause of death (death from prostate carcinoma vs. death from other causes) varied by age, race, clinical factors, and comorbid conditions. METHODS: A review was conducted of the medical records of decedent members of the Kaiser Permanente Medical Care program who previously were diagnosed with prostate carcinoma between January 1980 and December 1984 (n=584). The review focused on demographic factors, symptoms, diagnostic tests, stage of disease, and treatment. Data on comorbidity were obtained from a computerized discharge summary. Logistic regression analysis was used to estimate odds ratios. RESULTS: Approximately 54% of the decedent prostate carcinoma patients died of their prostate carcinoma. Decedents who were black, age < or = 65 years, diagnosed with more advanced disease stage, recipients of hormonal therapy, and whose death occurred > 6 months after diagnosis were more likely than others to die of prostate carcinoma. In contrast, the likelihood of dying of some other cause was associated with concurrent cardiovascular disease, after adjustment for the effects of race, age, and disease stage. There also were significant two-way age-race and age-time-to-death interactions. CONCLUSIONS: The prognostic significance of cardiovascular disease in prostate carcinoma patients should be investigated in subsequent survival studies. A number of questions need to be addressed delineating the complex relations between coexisting diseases and their treatment. PMID- 9740085 TI - Vascular pathology of malignant cervical lymphadenopathy: qualitative and quantitative assessment with power Doppler ultrasound. AB - BACKGROUND: Malignant vascular pathology has traditionally been studied with invasive angiography or in vitro immunohistochemistry. The objective of this study was to investigate the vascular patterns and vascular density of benign and malignant cervical lymphadenopathy using power Doppler ultrasound combined with a computed quantitative image processing system. METHODS: Investigations of 189 cervical lymph node lesions were undertaken prospectively using a 5-10 MHz linear array transducer in power mode. The types of vascular patterns displayed with power Doppler ultrasound, after sweep-scanning over the whole lymph node, were classified as hilar, spotted, peripheral, or mixed. Quantitative assessment of vascularity was made by sampling three parallel planes of each lymph node. A computed image processing system automatically calculated the density of vascular signals (called the "vascularity index" in this study) within the lymph node plane. RESULTS: Malignant lymph node lesions were shown to have higher vascularity indices (0.169+/-0.147, P < 0.01). The vascular patterns of benign lesions were mostly of avascular or hilar type (in 83% of cases). Malignant lesions were characterized by patterns of mixed (47%), spotted (20%), or peripheral type (11%). When vascular pattern (nonhilar type) and vascularity index (maximum > or = 0.09) were combined, the specificity for diagnosing malignant lymphadenopathy was as high as 97%. Variance in tumor vascularity was noted in both the benign and malignant groups. CONCLUSIONS: Power Doppler ultrasound combined with a computed image processing system provided an objective tool for assessing tumor vascularity quantitatively. Using this modality, the vascular pathology of malignant lymphadenopathy was found to be characterized by higher vascular density and aberrant vascular patterns. PMID- 9740086 TI - Malignant lymphoma in Thailand: changes in the frequency of malignant lymphoma determined from a histopathologic and immunophenotypic analysis of 425 cases at Siriraj Hospital. AB - BACKGROUND: Analysis of malignant lymphoma in a single institution at different periods of time can determine the changing status of the disease in the region. METHODS: To compare with the large series of 1095 lymphoma cases reported between 1957-1971 at Siriraj Hospital, the largest hospital in Thailand, a similar study was performed through histopathologic evaluation of 425 lymphoma cases diagnosed consecutively at the same institution between August 1993 and October 1995. Phenotypic analysis was performed by paraffin section-immunoperoxidase studies. RESULTS: A striking increase in lymphoma cases was noted from 73 cases/year in the first series to 189 cases/year in the second series (an increase of 158.9%). Lymphoma occurred in all age groups, with a peak incidence at the seventh decade of life. The male to female ratio decreased from 2:1 in 1957-1971 to 1.3:1 in the more recent series. The incidence of Hodgkin's disease (HD) was found to have decreased from 28.9% to 8.5%. There were 36 cases (8.5%) of HD and 389 cases (91.5%) of non-Hodgkin's lymphoma (NHL) reported in the second series. The subtypes of HD included 16 cases of mixed cellularity, 13 cases of nodular sclerosis, 6 cases of lymphocyte depletion, and 1 case of lymphocyte predominance. According to the Working Formulation, the 389 NHL cases included low grade (14.1%), intermediate grade (57.3%), high grade (11.3%), and miscellaneous groups (17.2%). They were classified as small lymphocytic (9.5%), follicular (11.1%), diffuse (50.9%), immunoblastic (4.1%), small noncleaved (4.4%), lymphoblastic (2.8%), anaplastic large cell (9.0%), mycosis fungoides (1.8%), hairy cell leukemia (0.3%), true histiocytic (0.5%), and extramedullary plasmacytoma (1.0%). The immunophenotypes of the 359 NHL cases available for paraffin section-immunoperoxidase studies were B-cell (71.0%), T-cell (24.5%), histiocyte (0.6%), and undetermined phenotypes (3.9%). CONCLUSIONS: The incidence of malignant lymphoma is increasing in Thailand, with a high frequency of intermediate to high grade NHL of B-cell phenotype reported. PMID- 9740087 TI - Long term treatment of myeloproliferative disease with interferon-alpha-2b: feasibility and efficacy. AB - BACKGROUND: Recombinant interferon-alpha-2b (rIFN-alpha-2b) has shown therapeutic potential in patients with chronic myelogenous leukemia and other myeloproliferative disorders (MPDs), including the ability to suppress the abnormal hematopoietic clone and to reverse myelofibrosis. This study was conducted to evaluate further the efficacy and safety of rIFN-alpha-2b in a large group of patients with polycythemia vera, essential thrombocythemia, or agnogenic myeloid metaplasia and to determine maintenance of response after treatment discontinuation. METHODS: Induction therapy began with subcutaneous rIFN-alpha-2b at 5.0 x 10(6) IU/day until a complete or partial response was achieved. Treatment continued at 2.5 x 10(6) IU/day until spleen size and hematologic parameters stabilized. RESULTS: Fifty-four patients were studied (median follow up, 7.3 years); at last follow-up 27 patients still were participating (median follow-up, 3.8 years). Twenty-four of 24 patients with thrombocythemia (100%) and 14 of 14 patients with hyperleukocytosis (100%) responded to induction therapy, whereas 26 of 39 patients (67%) experienced > 10% decrease in splenomegaly. Thirty-nine of 54 patients (72%) maintained response for a median of 39 weeks after withdrawal of rIFN-alpha-2b; repeat courses in previously responding patients produced similar results. The survival rate at 8 years was 60%. rIFN alpha-2b generally was well tolerated, but toxicity caused treatment withdrawal in 7 patients (13%). CONCLUSIONS: rIFN-alpha-2b can produce regression of splenomegaly and control of leukocyte and platelet counts in patients with MPD. These responses are sustained for prolonged periods in some patients after therapy discontinuation. In patients with recurrent disease, disease control can be attained again with reinitiation of rIFN-alpha-2b. Therefore this therapy should be an important treatment consideration for patients with MPD. PMID- 9740088 TI - Comparison of the efficacy and safety of tropisetron, metoclopramide, and chlorpromazine in the treatment of emesis associated with far advanced cancer. AB - BACKGROUND: A single institution, prospective, randomized trial was performed in terminal cancer patients to compare tropisetron (TRO), metoclopramide (MET), and chlorpromazine (CHL) in the management of nausea and emesis. Patients had far advanced cancer, were far removed from chemotherapy or radiotherapy, and their nausea and emesis was not due to bowel obstruction, drug intake, or cranial, electrolytic, or metabolic causes. The effects of antiemetic treatments were evaluated from Days 1-15. METHODS: Two hundred and eighty patients were randomized to receive 1) MET+ dexamethasone (DEX) (10 mg*4 and 2 mg*1, respectively, orally), 2) TRO (5 mg*1, orally), 3) TRO + MET (5 mg*1 and 10 mg*2, respectively, orally), 4) TRO + MET + DEX (5 mg*1, 10 mg*2, and 2 mg*1, respectively, orally), 5) CHL + DEX (25 mg*2 and 2 mg*1, respectively, orally), 6) TRO + CHL (5 mg*1 and 12.5 mg*2, respectively, orally), or 7) TRO + CHL + DEX (5 mg*1, 12.5 mg*2, and 2 mg*1, respectively, orally). Total control was defined as no nausea or emesis. RESULTS: By the end of the 15th day, total control of emesis was obtained in 23.6% (9 of 38) of MET + DEX patients, 78.9% (30 of 38) of TRO patients, 84.2% (32 of 38) of TRO + MET patients, 92.3% (36 of 39) of TRO + MET + DEX patients, 33.3 (13 of 39) of CHL + DEX patients, 84.6% (33 of 39) of TRO + CHL patients, and 92.5% (37 of 40) of TRO + CHL + DEX patients. Total control of nausea was achieved in 18.4% (7 of 38) of MET + DEX patients, 65.7% (25 of 38) of TRO patients, 73.6% (28 of 38) of TRO + MET patients, 87.1% (34 of 39) of TRO + MET + DEX patients, 17.9% (7 of 39) of CHL + DEX patients, 74.3% (29 of 39) of TRO + CHL patients, and 85% (34 of 40) of TRO + CHL + DEX patients. When comparing MET + DEX versus TRO; MET + DEX versus TRO + MET; MET + DEX versus TRO + MET + DEX; MET + DEX versus TRO + CHL; MET + DEX versus TRO + CHL + DEX; CHL + DEX versus TRO; CHL + DEX versus TRO + MET; CHL + DEX versus TRO + MET + DEX; CHL + DEX versus TRO + CHL; and CHL + DEX versus TRO + CHL + DEX, significant differences were noted. All antiemetic drugs were well tolerated with no severe side effects observed in any treatment arm. CONCLUSIONS: These data suggest that 5-HT3 receptor antagonists such as tropisetron clinically are more effective in the control of emesis of patients with far advanced cancer than previously used agents. This study raises important issues when attempting to decide which antiemetic therapy to choose for an individual patient with far advanced disease. PMID- 9740089 TI - Liver dysfunction in patients infected with hepatitis C virus undergoing chemotherapy for hematologic malignancies. AB - BACKGROUND: Reactivation of chronic hepatitis B virus (HBV) infection with the development of fulminant hepatitis induced by withdrawal of chemotherapy and/or corticosteroids (CS) is well known. However, less is known about liver dysfunction in patients with hepatitis C virus (HCV) who are undergoing chemotherapy. Thus, the authors conducted this study to determine whether chemotherapy for HCV positive patients with hematologic malignancies is associated with hepatic injury. METHODS: Thirty-three consecutive HCV positive patients were studied. Twenty-six had B-cell lymphoma, two had Hodgkin's disease, two had acute myeloblastic leukemia (AML), two had chronic myelocytic leukemia, and one had multiple myeloma. HCV infection was detected by anti-HCV antibodies (enzyme immunoassay) and HCV RNA (reverse transcription polymerase chain reaction). In 28 of 33 patients, CS were used as part of the chemotherapy regimens. Liver function tests (LFTs) were evaluated prior to chemotherapy, a mean of 19 days after each cycle of chemotherapy, and during the follow-up period after the completion of chemotherapy. Mean follow-up was 14 months (range, 7-26 months). RESULTS: Twenty-seven of 33 patients (82%) were positive for both anti HCV and HCV RNA, 5 for HCV RNA only, and 1 for anti-HCV antibodies only. Fourteen patients (42%) had normal pretreatment LFTs. During treatment, 18 patients (55%) (7 with normal and 11 with abnormal pretreatment transaminase levels) had mild-to moderate increases of alanine aminotransferase (ALT) (median, 156 U/L; range, 59 491) and aspartate aminotransferase (AST) (median, 121; range, 45-243), which occurred 2-3 weeks after the withdrawal of chemotherapy without associated hyperbilirubinemia. Only one patient with baseline ALT and AST of 182 IU/L and 117 IU/L had a severe "flare" of hepatitis C, with peak ALT and AST of 491 IU/L and 243 IU/L. No patient developed fulminant hepatitis or died of liver-related causes. Posttreatment levels of transaminases were not significantly different from pretreatment levels. Abnormal pretreatment transaminase levels did not predict further increase during treatment. CONCLUSIONS: Significant hepatic dysfunction is uncommon among HCV infected patients treated with chemotherapy for hematologic malignancies. PMID- 9740090 TI - Idiopathic giant cell myocarditis after autologous hematopoietic stem cell transplantation and interleukin-2 immunotherapy: a case report. AB - BACKGROUND: Interleukin-2 (IL-2) is used in the treatment of solid tumors and hematologic malignancies. Sudden death is a rare complication of IL-2 treatment. METHODS: A patient with lymphoma underwent chemoradiotherapy myeloablation and autologous stem cell transplantation. The stem cells were cultured in IL-2 (6000 IU/mL) for 24 hours prior to infusion. After engraftment, treatment with IL-2 (1.8 x 10(6) IU/m2/day administered subcutaneously) was begun. After 4 days of treatment, the patient suddenly died. An autopsy was performed. RESULTS: Histologic examination of the myocardium revealed a diffuse, lymphocytic infiltrate with scattered, multinucleated giant cells and foci of myocardial degeneration consistent with giant cell myocarditis. The lymphocytes were predominantly CD4 positive T cells, and the majority of these cells stained with antibodies for perforin, suggesting an unusual cytolytic role for these lymphocytes. DNA end-labeling of myocardial tissue sections revealed numerous apoptotic myocytes within the lymphocytic infiltrate. CONCLUSIONS: To the authors' knowledge, this is the first report of giant cell myocarditis in association with high dose chemotherapy, transplantation, and IL-2 immunomodulation. The authors suggest that the cytokine imbalance produced by IL 2 may have initiated a preferential activation of T helper cells and an autoimmune phenomenon manifesting as giant cell myocarditis. PMID- 9740091 TI - Liver transplant recipients are not at increased risk for nonlymphoid solid organ tumors. AB - BACKGROUND: Organ transplant recipients are at higher risk for developing lymphoid tumors, skin carcinomas, and sarcomas. Whether liver transplant recipients are at higher risk for developing more common cancers is unclear. METHODS: All patients with a history of malignancy prior to liver transplantation and those who developed malignancy, either de novo or recurrent, after transplantation were identified retrospectively. The following parameters were examined: age at diagnosis; indication for transplant; interval from transplant to tumor diagnosis; tumor treatment received; predisposing factors for the development of cancer; immunosuppression regimen, including the use of OKT3; number and treatment of rejection episodes; and survival. RESULTS: Of 888 patients, 29 (3.2%) had 31 previous malignancies; of these 29 patients, 4 developed a recurrence in the posttransplant period. Thirty-nine patients (4.3%) developed 43 de novo nonlymphoid malignancies. Alcoholic cirrhotic patients had a significantly higher incidence of de novo carcinomas. Except for skin carcinomas, tumors did not occur with greater frequency than in the general population, and recurrent tumors were not more aggressive than reported for that disease. One patient had an unrecognized renal cell carcinoma at the time of transplant that progressed rapidly; this patient died 64 days after transplantation. CONCLUSIONS: With current immunosuppressive regimens, liver transplant patients do not appear to be at an increased risk for developing nonlymphoid solid organ tumors. However, longer follow-up will be necessary to confirm these results. PMID- 9740092 TI - Age, pain intensity, and opioid dose in patients with advanced cancer. AB - BACKGROUND: Elderly patients are more likely to be affected by the acute and chronic toxicities of opioids, but an association between age and long term opioid consumption has not been established clearly in patients with advanced cancer. METHODS: The computerized records of 197 cancer patients admitted to a palliative care unit in Edmonton, Alberta, Canada were examined. The authors examined: demographics (age, gender, and location of primary tumor), pain characteristics (presence of neuropathic pain and incidental pain), mean daily pain intensity (MDPI), and daily opioid consumption measured as (parenteral) morphine equivalent daily dose (MEDD). MDPI and MEDD were assessed on Days 2 and 7 after admission, on the day of maximum opioid consumption, and on the day of maximum pain intensity during admission. Average values for MDPI and MEDD were calculated between Days 2 and 7. RESULTS: When age was treated as a categoric variable (< 65 years, 65-74 years, and 75+ years), statistically significant differences in MEDD were observed for age for all estimates except those for Day 7, with older patients requiring a lower equianalgesic dose. No major differences were observed for pain intensity and for the presence of incidental or neuropathic pain across the different age groups. In the multivariate analysis, the reduction in MEDD ranged between 27-71 mg when patients age > or = 75 years were compared with younger adults. A MEDD increase that ranged between 82-137 mg was associated with the presence of neuropathic pain. CONCLUSIONS: The current study suggests that elderly cancer patients may experience a similar level of pain intensity but require a lower amount of opioid analgesia than younger adults. However, because elderly patients are more likely to be affected by the acute and chronic toxicities of opioids, opioids should be initially administered at a lower dose and titrated cautiously in these patients. PMID- 9740093 TI - A phase I trial of continuous hyperthermic peritoneal perfusion with tumor necrosis factor and cisplatin in the treatment of peritoneal carcinomatosis. AB - BACKGROUND: Tumor necrosis factor (TNF), hyperthermia, and cisplatin have synergistic cytotoxicity against cancer cells in vitro. This combination may be well suited to the regional treatment of peritoneal tumor spread in patients. Continuous hyperthermic peritoneal perfusion (CHPP) is a technique that allows uniform delivery of cytotoxic agents and heat to the peritoneal surface. A Phase I trial of CHPP with TNF and cisplatin was conducted to define the maximum tolerated dose (MTD) for TNF and cisplatin under moderate hyperthermia in the treatment of peritoneal carcinomatosis. METHODS: Twenty-seven patients with peritoneal carcinomatosis underwent exploratory laparotomy and tumor debulking followed by a 90-minute CHPP with cisplatin (100-350 mg/m2) and TNF (0-0.3 mg/L). Perfusion parameters included a perfusate volume of 3-9 L, a peritoneal temperature of 42-43 degrees C, and a flow rate of 1.5 L/minute. Sodium thiosulfate was administered systemically during and after the perfusion as a cisplatin binding agent. RESULTS: There was no operative or treatment-related mortality in this study. CHPP resulted in a 14-fold higher area under the concentration versus time curve (AUC) for cisplatin in the perfusate compared with plasma, and a 4854-fold higher AUC for TNF. The MTD was defined as 250 mg/m2 cisplatin plus 0.1 mg/L TNF. The dose-limiting toxicity was renal insufficiency. No other systemic toxicity was identified, and no significant regional toxicity was identified. The median time to toleration of a regular diet was 8 days (range, 5-20 days). CONCLUSIONS: The favorable regional pharmacologic profile of the combination of cisplatin and TNF suggests that these agents administered via CHPP warrant further evaluation as prophylaxis against or treatment for peritoneal carcinomatosis. PMID- 9740094 TI - The National Cancer Data Base 10-year survey of breast carcinoma treatment at hospitals in the United States. AB - BACKGROUND: The National Cancer Data Base (NCDB), a joint project of the American College of Surgeons Commission on Cancer and the American Cancer Society, is a cancer management and outcome data base for health care organizations. It provides a comparative summary of patient care that is used by participating hospitals and communities for self-assessment. The most current (1995) data are described herein. METHODS: Since 1989, seven calls for data have been issued, yielding reports on a total of 240,031 breast carcinoma patients for the years included in this analysis. A total of 1849 hospital cancer registries responded to at least 1 of the calls for data. RESULTS: A continuous improvement in care was reported. By 1995, 45.8% (nearly one-half) of breast carcinoma patients were diagnosed early as Stage 0 or I, and early stage patients (Stage 0 or I) were most often treated with partial mastectomy (in 58% of cases). Favorable 10-year relative survival rates for Stage 0 (95%) and Stage I (88%) breast carcinoma patients were reported. Patients who were presumed to be Stage I and were not selected for axillary dissection had poorer survival. Survival differences were reported for different treatment groups within individual stage strata. Over the 10-year observation period, fewer patients from lower-income neighborhoods were diagnosed with early stage breast carcinoma. In general, the annual relative survival rate remained constant over the 10-year observation period (with no plateau after 5 years) within each stage and for all stages combined. CONCLUSIONS: Improvements in diagnosis and treatment during the period 1985-1995 were demonstrated by these data. The NCDB breast carcinoma data are appropriate norms for formal quality assurance purposes, such as those specified by the Standards of the Commission on Cancer published by the American College of Surgeons Commission on Cancer. Cancer committees and other clinicians working within the hospital setting should assess and compare stage distribution, stage specific treatment patterns, and the correlations between the outcomes of patients and both disease stage and treatment. PMID- 9740095 TI - Evaluation of focal cerebral ischemia in rats by magnetic resonance imaging and immunohistochemical analyses. AB - Correlation of focal ischemia-induced brain damage evidenced by magnetic resonance imaging (MRI) and by staining with microtubule-associated protein 2 (MAP2) was studied in rats. Ischemia was produced by transient occlusion of the middle cerebral artery (MCAO). The damage was assessed at 6 to 8 hours after MCAO and 1 week later. The area of damage assessed by MRI agreed with that by MAP2 staining at 6 to 8 hours after MCAO, which was smaller (P < 0.001) than that defined by MAP2 staining 1 week after MCAO. Glial staining indicated that glial infiltration affected the signal intensity of MRI in the area of damage. PMID- 9740096 TI - Cerebral blood flow measurements by magnetic resonance imaging bolus tracking: comparison with [(15)O]H2O positron emission tomography in humans. AB - In six young, healthy volunteers, a novel method to determine cerebral blood flow (CBF) using magnetic resonance (MR) bolus tracking was compared with [(15)O]H2O positron emission tomography (PET). The method yielded parametric CBF images with tissue contrast in good agreement with parametric PET CBF images. Introducing a common conversion factor, MR CBF values could be converted into absolute flow rates, allowing comparison of CBF values among normal subjects. PMID- 9740097 TI - Comparison of the transient equilibrium and continuous infusion method for quantitative PET analysis of [11C]raclopride binding. AB - Several approaches have been applied for quantification of D2 dopamine receptors in positron emission tomography studies using [11C]raclopride. Initial approaches were based on analyses of data obtained after rapid bolus injection of [11C]raclopride. A continuous infusion paradigm has more recently been applied. The current study compares these approaches in healthy men. Two positron emission tomography measurements were performed in each of six healthy men, the first with rapid bolus injection and the second with continuous infusion of [11C]raclopride. In rapid bolus injection, the binding potential was calculated by the following methods. One approach is the kinetic analysis using the standard three compartment model. Another is to define a transient equilibrium at the moment when the specific binding reaches its maximum. In continuous infusion, binding potential was calculated by using time-activity data at equilibrium condition. All methods gave almost identical binding potential, representing cross validation of these methods. The continuous infusion method can provide "true" equilibrium condition. The kinetic analysis is a sophisticated approach but requires determination of an arterial input function. The transient equilibrium method thus is suitable for routine clinical research, since it does not require determination of an arterial input function. PMID- 9740098 TI - Validation of the three-dimensional acquisition mode in positron emission tomography for the quantitation of [18F]fluoro-DOPA uptake in the human striata. AB - Three-dimensional (3D) positron emission tomography (PET) is attractive for [18F]fluoro-DOPA studies, since the sensitivity improvement is maximal for radioactive sources located in central planes, which is usually the case for the human striata. However, the image quantitation in that mode must be assessed because of the nearly threefold increase in scattered coincidences. We report the results of [18F]fluoro-DOPA studies performed on six normal volunteers. Each one was scanned in the 3D and two-dimensional (2D) modes on the same tomograph. The quantitation in the 3D and 2D modes was compared for a Patlak graphical analysis with the occipital counts as the input function (Ki) and a striatooccipital ratio analysis. We find that, in 3D PET, a scatter correction is required to preserve the same quantitation as in 2D PET. When the 3D data sets are corrected for scatter, the quantitation of the [18F]fluoro-DOPA uptake, using the Patlak analysis, is similar in the 2D and 3D acquisition modes. Conversely, analysis of the striatooccipital ratio leads to higher values in 3D PET because of a better in-plane resolution. Finally, using the 3D mode, the dose injected to the subjects can be reduced by a factor greater than 1.5 without any loss in accuracy compared to the 2D mode. PMID- 9740099 TI - Neural activation of the brain with hemodynamic insufficiency. AB - Little is known about how ischemia affects hemodynamic responses to neural activation in the brain. We compare the effects of a motor activation task and a cerebral vasodilating agent, acetazolamide (ACZ), on regional cerebral blood flow (rCBF) in primary sensorimotor cortex (PSM) in six patients with major cerebral artery steno-occlusive lesions without paresis of the upper extremities. Quantitative rCBF was measured in all patients using H2(15)O autoradiographic method and positron emission tomography. The CBF was determined at rest, during a bimanual motor activation task, and 10 minutes after ACZ administration. With bimanual motor activation, rCBF increased significantly in both PSM compared with at rest (P < 0.01 on lesion side, and P < 0.02 on contralateral side). However, rCBF did not increase after ACZ injection in the PSM on the lesion side, whereas rCBF increased significantly in the contralateral PSM after ACZ injection compared with the level at rest. This result suggests that despite a decreased hemodynamic reserve, there is a nearly normal flow response to neural activation, indicating that the mechanism of vasodilation responsible for perfusion change is different for acetazolamide and neural activation. The relations among neural activation, hemodynamic status, and cerebral metabolism in the ischemic stroke patients are discussed. PMID- 9740101 TI - Nitric oxide scavenging by hemoglobin or nitric oxide synthase inhibition by N nitro-L-arginine induces cortical spreading ischemia when K+ is increased in the subarachnoid space. AB - We investigated the combined effect of increased brain topical K+ concentration and reduction of the nitric oxide (NO.) level caused by nitric oxide scavenging or nitric oxide synthase (NOS) inhibition on regional cerebral blood flow and subarachnoid direct current (DC) potential. Using thiopental-anesthetized male Wistar rats with a closed cranial window preparation, brain topical superfusion of a combination of the NO. scavenger hemoglobin (Hb; 2 mmol/L) and increased K+ concentration in the artificial cerebrospinal fluid ([K+]ACSF) at 35 mmol/L led to sudden spontaneous transient ischemic events with a decrease of CBF to 14+/-7% (n=4) compared with the baseline (100%). The ischemic events lasted for 53+/-17 minutes and were associated with a negative subarachnoid DC shift of -7.3+/-0.6 mV of 49+/-12 minutes' duration. The combination of the NOS inhibitor N-nitro-L arginine (L-NA, 1 mmol/L) with [K+]ACSF at 35 mmol/L caused similar spontaneous transient ischemic events in 13 rats. When cortical spreading depression was induced by KCl at a 5-mm distance, a typical cortical spreading hyperemia (CSH) and negative DC shift were measured at the closed cranial window during brain topical superfusion with either physiologic artificial CSF (n=5), or artificial CSF containing increased [K+]ACSF at 20 mmol/L (n=4), [K+]ACSF at 3 mmol/L combined with L-NA (n=10), [K+]ACSF at 10 mmol/L combined with L-NA (five of six animals) or [K+]ACSF at 3 mmol/L combined with Hb (three of four animals). Cortical spreading depression induced longlasting transient ischemia instead of CSH, when brain was superfused with either [K+]ACSF at 20 mmol/L combined with Hb (CBF decrease to 20+/-20% duration 25+/-21 minutes, n=4), or [K+]ACSF at 20 mmol/L combined with L-NA (n=19). Transient ischemia induced by NOS inhibition and [K],ACSF at 20 mmol/L propagated at a speed of 3.4+/-0.6 mm/min, indicating cortical spreading ischemia (CSI). Although CSH did not change oxygen free radical production, as measured on-line by in vivo lucigenin-enhanced chemiluminescence, CSI resulted in the typical radical production pattern of ischemia and reperfusion suggestive of brain damage (n=4). Nimodipine (2 microg/kg body weight/min intravenously) transformed CSI back to CSH (n=4). Vehicle had no effect on CSI (n=4). Our data suggest that the combination of decreased NO. levels and increased subarachnoid K+ levels induces spreading depression with acute ischemic CBF response. Thus, a disturbed coupling of metabolism and CBF can cause ischemia. We speculate that CSI may be related to delayed ischemic deficits after subarachnoid hemorrhage, a clinical condition in which the release of Hb and K+ from erythrocytes creates a microenvironment similar to the one investigated here. PMID- 9740100 TI - Optical intrinsic signal imaging responses are modulated in rodent somatosensory cortex during simultaneous whisker and forelimb stimulation. AB - Optical intrinsic signal imaging (OIS) was used to investigate physiologic interactions between spatially and functionally distinct cortical somatosensory systems. The OIS response magnitude was evaluated after simultaneous stimulation of single whiskers and forelimb digits. Whisker C1 was deflected at a frequency of 10 Hz for 2 seconds while low- or high-intensity vibratory stimuli were applied to forelimb digits. The OIS responses to simultaneous whisker and forelimb stimulation were compared with lone whisker stimulated controls. Overall, addition of a second stimulus caused decreases in barrel cortex response magnitude. Three different response patterns were detected within individual trial sets. Modulation of barrel cortex evoked potentials provided evidence that changes in OIS responses observed here may be partially influenced by vascular responses to changes in neuronal activity. However, OIS responses in the barrel region during lone forelimb stimulation that were unaccompanied by evoked potentials suggested the possibility of independent vascular dynamic influences on response modulation. This study demonstrates that cortical responses at the level of primary sensory processing may be significantly influenced by activity in adjacent regions. Furthermore, it reveals that vascular and neuronal characteristics of interregional modulation do not co-localize and may produce responses in which one component increases while the other decreases. PMID- 9740102 TI - Enhanced poly(ADP-ribosyl)ation after focal ischemia in rat brain. AB - Nitric oxide from neuronal cells plays detrimental roles in glutamate neurotoxicity and in focal brain ischemia. Nitric oxide directly damages DNA, and breaks in the DNA strands activate poly(ADP-ribose) polymerase (PARP), which brings poly(ADP-ribosyl)ation of the nuclear proteins. The excessive activation of PARP is thought to cause depletion of ATP and the energy failure resulting in cell death. To clarify the involvement of poly(ADP-ribosyl)ation in ischemic insult, we examined poly(ADP ribosyl)ation by immunohistochemical methods and the protective effect of 3-aminobenzamide, which is a PARP inhibitor, on focal brain ischemia using an intraluminal permanent middle cerebral artery occlusion model in rats. Poly(ADP ribosyl)ation was widely and markedly detected 2 hours after the ischemic insult in the cerebral cortex and striatum in which infarction developed 24 hours later. The enhanced immunoreactivity of poly(ADP-ribose) gradually decreased, and 16 hours later, no immunoreactivity was detected. Intraventricular administration of 3-aminobenzamide (1 to 30 mg/kg) 30 minutes before the ischemic insult decreased infarction volume in a dose-dependent manner along with the immunohistochemical reduction of poly(ADP-ribosyl)ation. Pretreatment with 7-nitroindazole (25 mg/kg, intraperitoneally), a selective neuronal nitric oxide synthetase inhibitor, partially reduced poly(ADP ribosyl)ation. These data suggest the involvement of poly(ADP-ribosyl)ation in the development of cerebral infarction. PMID- 9740103 TI - Spreading depression in focal ischemia: a computational study. AB - When a cerebral infarction occurs, surrounding the core of dying tissue there usually is an ischemic penumbra of nonfunctional but still viable tissue. One current but controversial hypothesis is that this penumbra tissue often eventually dies because of the metabolic stress imposed by multiple cortical spreading depression (CSD) waves, that is, by ischemic depolarizations. We describe here a computational model of CSD developed to study the implications of this hypothesis. After simulated infarction, the model displays the linear relation between final infarct size and the number of CSD waves traversing the penumbra that has been reported experimentally, although damage with each individual wave progresses nonlinearly with time. It successfully reproduces the experimental dependency of final infarct size on midpenumbra cerebral blood flow and potassium reuptake rates, and predicts a critical penumbra blood flow rate beyond which damage does not occur. The model reproduces the dependency of CSD wave propagation on N-methyl-D-aspartate activation. It also makes testable predictions about the number, velocity, and duration of ischemic CSD waves and predicts a positive correlation between the duration of elevated potassium in the infarct core and the number of CSD waves. These findings support the hypothesis that CSD waves play an important causal role in the death of ischemic penumbra tissue. PMID- 9740104 TI - Magnetic resonance imaging assessment of cerebral hemodynamics during spreading depression in rats. AB - High-speed magnetic resonance imaging was used to perform simultaneous measurements of relative cerebral blood volume (rCBV) and water diffusion changes during spreading depression (SD) induced by cortical potassium chloride application. Rats were fitted epidurally with a rubber chamber. Potassium chloride was perfused through the chamber until SD was indicated by a negative direct current (DC) potential shift. Magnetic resonance imaging scans used echo planar diffusion and T2-weighted images. Iron dextran was injected as a blood pool contrast agent to make subsequent changes in T2 (or T2*) directly proportional to changes in CBV. Multislice maps of apparent diffusion coefficient (ADC) and rCBV were generated with 6- to 16-second time resolution, which revealed transient ADC and rCBV changes propagating over the cortex after potassium chloride application. Transient ADC declines appeared simultaneously with the DC shift, whereas rCBV increase followed with a delay of 16.4+/-14.9 seconds. Prolonged rCBV decrease was observed after the initial increase during the SD in half of the animals. The delayed rCBV response after the ADC change supports the observation of increased energy demand because of repolarization. Simultaneous DC potential recording and ADC measurements in corresponding sites of the cortex indicate that transient ADC decreases during SD reflect water shifts associated with cell depolarization. PMID- 9740105 TI - Experimental hypoxemic hypoxia: effects of variation in hematocrit on magnetic resonance T2*-weighted brain images. AB - T2*-weighted gradient echo magnetic resonance images of rat brain were obtained dynamically during acute hypoxemic hypoxia to investigate the relations between changes in cerebral blood oxygen saturation (deltaYb), blood hematocrit (Hct), and R2* (deltaR2*). Images from hypoxemic rats with normal Hct (42.8%+/-2.33%; n=12) were compared with those from hypoxemic rats with mild (33.4%+/-1.88%; n=8) or moderate (27.14%+/-2.7%; n=10) reduction of Hct. A linear relation between deltaYb and deltaR2* was obtained for all three groups. However, the slopes of the linear regressions were statistically different from one another (P < 0.001), with the slopes of the regression lines increasing inversely with Hct; that is, the slope for normal Hct is less than the slope for mildly reduced Hct, which is less than the slope for moderately reduced Hct. These data suggest that for any given reduction in the oxygen saturation of cerebral blood, the deltaR2* will be of a lesser magnitude when the hemoglobin concentration is reduced; the data are consistent with existing theoretical models of deoxyhemoglobin content-dependent effects in T2*-weighted magnetic resonance imaging. PMID- 9740106 TI - Polynitroxyl albumin reduces infarct size in transient focal cerebral ischemia in the rat: potential mechanisms studied by magnetic resonance imaging. AB - Nitroxide free radicals are known to protect cells from oxidative damage. Diffusion-weighted and perfusion-weighted magnetic resonance imaging was used to evaluate the effects of polynitroxyl albumin (PNA) in a middle cerebral artery intraluminal suture model of transient focal cerebral ischemia in the rat. Three groups of Sprague-Dawley rats were investigated: (1) PNA (N=6), (2) human serum albumin (N =6), and (3) saline (N=7). The middle cerebral artery was occluded for 2 hours. Treatment was started 30 minutes after induction of ischemia. A total dose of 1% body weight (volume/weight) of PNA (23.5 mg/dL protein and 110 mmol/L nitroxide), albumin (23.5 mg/dL), or saline was injected intravenously at three time points: 0.5% at 0.5 hours, 0.25% at 2 hours (i.e., just before reperfusion), and 0.25% at 4 hours after occlusion. Six sets of diffusion- and perfusion weighted magnetic resonance images were acquired throughout the 2 hours of ischemia and the 2 hours of reperfusion. The rats were killed at 24 hours, and the brains were stained with 2,3,5-triphenyltetrazolium chloride (TTC). Diffusion weighted imaging showed that the growth of the ischemic lesion was suppressed in the PNA-treated group. The 4 hours diffusion-weighted imaging--derived hemispheric lesion volume in the PNA-treated group (25%+/-9%) was significantly smaller than that in the saline-treated (43%+/-13%; P=0.016) or albumin-treated groups (38%+/-6%; P=0.017). A larger difference was observed for the 24-hour TTC derived lesion volumes in the PNA (8%+/-7%), saline (35%+/-8%; P < 0.001), and albumin (31%+/-6%; P < 0.001) groups. Perfusion-weighted imaging demonstrated a marked improvement in cerebral perfusion in the PNA-treated group during ischemia and reperfusion. In conclusion, treatment with PNA results in an improvement in perfusion and a reduction of infarct volume in a model of transient focal cerebral ischemia in the rat. PMID- 9740107 TI - Stimulation of beta2-adrenoceptors inhibits apoptosis in rat brain after transient forebrain ischemia. AB - We have previously demonstrated that the neuroprotective effect of the beta2 adrenoceptor agonist clenbuterol in vitro and in vivo was most likely mediated by an increased nerve growth factor (NGF) expression. In the present study, we examined whether clenbuterol was capable of inhibiting apoptosis caused by ischemia. Transient forebrain ischemia was performed in male Wistar rats (300 to 350 g) by clamping both common carotid arteries and reducing the blood pressure to 40 mm Hg for 10 minutes. Clenbuterol (0.1, 0.5, and 1.0 mg/kg intraperitoneally) was administered 3 hours before ischemia or immediately after ischemia. The brains were removed for histologic evaluation 7 days after ischemia. The time course of DNA fragmentation was determined 1, 2, 3 and 4 days after ischemia. Staining with terminal deoxynucleotidyl transferase (TdT) mediated dUTP nick end-labeling (TUNEL) was used for further analysis of DNA fragments in situ 3 days after ischemia. The NGF protein was assayed by enzyme linked immunosorbent assay. Ten-minute forebrain ischemia damaged 80% to 90% of the neurons in the hippocampal CA1 region evaluated 7 days after ischemia. Pretreatment with clenbuterol (0.5 and 1.0 mg/kg) reduced the neuronal damage by 18.1% (P < 0.01) and 13.1% (P < 0.05), respectively. The neuroprotective effect also was found when clenbuterol (0.5 mg/kg) was administered immediately after ischemia (P < 0.05). The DNA laddering appeared in striatum 1 day and in hippocampus 2 days after ischemia and peaked on the third day in both regions. The DNA laddering was nearly abolished in the hippocampus and partially blocked in striatum and cortex by 0.5 mg/kg clenbuterol. These results were confirmed by TUNEL staining. Clenbuterol (0.5 mg/kg intraperitoneally) elevated the NGF protein level by 33% (P < 0.05) in the hippocampus and 41% (P < 0.05) in the cortex 6 hours after ischemia. Three days after ischemia, the NGF levels in these regions were no longer different between the clenbuterol-treated and control groups. This study clearly demonstrates that clenbuterol possesses a neuroprotective activity and a marked capacity to inhibit DNA degradation after global ischemia. The results suggest that clenbuterol increases NGF expression during the first hours after global ischemia and thereby protects neurons against apoptotic damage. PMID- 9740108 TI - Stimulation of tyrosine phosphorylation of a brain protein by hibernation. AB - Mammalian hibernation is a state of natural tolerance to severely decreased brain blood flow. As protein tyrosine phosphorylation is believed to be involved in the development of resistance to potentially cell-damaging insults, we used immunoblotting for the phosphotyrosine moiety to analyze extracts from various tissues of hibernating and nonhibernating ground squirrels. A single, hibernation specific phosphoprotein was detected in the brain, but not in any other tissue tested. This protein, designated pp98 to reflect its apparent molecular weight, is distributed throughout the brain, and is associated with the cellular membrane fraction. The presence of the protein is tightly linked to the hibernation state; it is not present in contemporaneously assayed animals that are exposed to the same cold temperature as the hibernators, is present for the duration of a hibernation bout (tested from 1 to 14 days), and disappears within 1 hour of arousal from hibernation. The close association of pp98 with the hibernation state, its presence in cellular membranes, and the known properties of membrane phosphotyrosine proteins suggest that it may transduce a signal for adaptation to the limited availability of oxygen and glucose and low cellular temperature that characterizes hibernation in the ground squirrel. PMID- 9740109 TI - New neuropathological criteria for Alzheimer disease. PMID- 9740110 TI - Alzheimer disease-related abnormalities of amyloid beta precursor protein isoforms in the platelet: the brain's delegate in the periphery? PMID- 9740111 TI - New antiepileptic drugs. AB - The current developments in the availability of new antiepileptic drugs (AEDs) are unprecedented. After a period of many years during which no new AED became available, 5 new AEDs were introduced in the United States between 1993 and 1997, and 2 more are expected to be approved soon. These new drugs are a most welcome addition to the therapeutic options in the treatment of epilepsy, but they also create a dilemma for the clinician because their individual places and their optimal use in the treatment of various forms of epilepsy are yet to be determined. This review serves to summarize the main characteristics of the newer AEDs. PMID- 9740112 TI - Regional distribution of neuritic plaques in the nondemented elderly and subjects with very mild Alzheimer disease. AB - BACKGROUND: Identification of the neuropathological lesions that are most closely associated with the earliest symptoms of Alzheimer disease (AD) is crucial to the understanding of the disease process and the development of treatment strategies to affect its progress. Do the classical neuropathological lesions of AD precede, follow, or occur in synchrony with the earliest signs of cognitive deterioration? DESIGN AND OUTCOME MEASURES: We examined the extent of neuritic plaque (NP) formation in 5 neocortical regions and the hippocampus, entorhinal cortex, and amygdala in 66 elderly subjects with no dementia, questionable dementia, or mild dementia as assessed using the Clinical Dementia Rating Scale (CDR). SETTING AND PATIENTS: Postmortem study of nursing home residents. RESULTS: Even questionable dementia (CDR, 0.5) was associated with a significant (P = .04) increase in neocortical NP density. The density of NPs increased further with increasing dementia severity in all brain regions examined. However, subjects with questionable dementia or definite but mild dementia did not differ significantly from each other. Density of NPs was nearly maximal in subjects with moderate dementia (CDR = 2.0), suggesting that other neuropathological changes may be responsible for cognitive deficits beyond this level. Dementia severity correlated significantly with the density of NPs in all brain regions examined (r range, 0.47-0.56; P < .001), even when subjects with a CDR of 0 were excluded. CONCLUSIONS: These findings are consistent with the hypothesis that NPs are among the earliest neuropathological lesions in AD. Even very mild or questionable dementia is associated with increased density of neocortical NPs that do not distinguish between clinically questionable vs definite dementia. PMID- 9740113 TI - Differential level of platelet amyloid beta precursor protein isoforms: an early marker for Alzheimer disease. AB - OBJECTIVE: To determine whether a differential level of platelet amyloid beta precursor protein (APP) isoforms is specifically related to Alzheimer disease (AD) and whether it shows a correlation with the progression of clinical symptoms. DESIGN: After subjects were grouped according to diagnosis and severity of dementia, APP isoform levels in platelets were compared. SETTING: University medical centers. PATIENTS: Thirty-two patients who fulfilled diagnostic criteria for probable AD, 25 age-matched control subjects, and 16 patients with non-AD dementia. MAIN OUTCOME MEASURE: The levels of APP isoforms were evaluated by means of Western blot analysis and immunostaining of whole platelets. Messenger RNAs for APP transcripts were also evaluated by means of reverse transcriptase polymerase chain reaction. RESULTS: The ratio between the intensity of the 130-kd and 106- to 110-kd APP isoforms was significantly lower in the AD group (0.31 +/- 0.15, mean +/- SD) compared with both controls (0.84 +/- 0.2) and non-AD subjects (0.97 +/- 0.4). The ratio of platelet APP isoforms in patients with AD grouped by Clinical Diagnostic Rating score significantly correlated with the severity of the disease (Pearson correlation coefficient, followed by Bonferroni correction, P = .01). Reverse transcriptase polymerase chain reaction experiments showed that APP transcripts in all experimental groups were equally expressed. CONCLUSIONS: The pattern of platelet APP isoforms is specifically altered in patients with AD. In addition, the alteration of platelet APP isoforms shows a positive correlation with the progression of clinical symptoms, supporting the possibility to consider this peripheral parameter as a marker of progression of the disease. These alterations are not related to abnormalities of APP isoforms messenger RNAs in platelets. PMID- 9740114 TI - Motor, cognitive, and behavioral performance following unilateral ventroposterior pallidotomy for Parkinson disease. AB - OBJECTIVE: To evaluate the effects of ventroposterior pallidotomy on motor disability and on behavior and cognition in patients with medically intractable idiopathic Parkinson disease. DESIGN: Detailed motor testing both while receiving and discontinuing levodopa medication, posturography, and neurocognitive and behavioral assessments were performed before and 3 to 6 months after unilateral ventroposterior pallidotomy. SETTING: University-based movement disorder program. PATIENTS: Thirty-two patients without dementia with medically refractory idiopathic Parkinson disease were studied. MAIN OUTCOME MEASURES: Motor function and disability were measured using the Unified Parkinson's Disease Rating Scale, Hoehn and Yahr stage, and the Schwab and England Activities of Daily Living Scale. Dynamic balance was measured by sway (amplitude and velocity) using the Chattecx Balance System. Detailed cognitive and behavioral assessments were also performed both before and after surgery. RESULTS: Eighty-three percent of patients experienced improvement of their total Unified Parkinson's Disease Rating Scale score at 3 to 6 months after surgery. Significant improvements were also seen in the contralateral Unified Parkinson's Disease Rating Scale motor subscore (78%) as well as in the contralateral Unified Parkinson's Disease Rating Scale total score both during the on and off period (78% and 79%, respectively). The Hoehn and Yahr stage, Schwab and England Activities of Daily Living Scale score, and dynamic balance when standing on foam also improved following unilateral pallidotomy in many patients. Cognitive performance remained relatively unchanged following surgery with the exception of category fluency, which exhibited a modest decline (P < .04). A significant improvement in depression was found on the Beck Depression Inventory. CONCLUSIONS: Ventroposterior pallidotomy significantly improves motor performance and daily level of function in Parkinson disease. Cognition and behavior are not adversely affected in patients without dementia, and a cognitive screening battery is proposed. PMID- 9740115 TI - Ropinirole for the treatment of early Parkinson disease: a 12-month experience. Ropinirole Study Group. AB - OBJECTIVE: To evaluate ropinirole hydrochloride as dopaminergic monotherapy in patients with early Parkinson disease. DESIGN: A 6-month extension of a double blind, placebo-controlled study. SETTING: Ambulatory care at 22 different sites in the United States. PATIENTS: Patients who successfully completed the initial 6 month study could enter the 6-month extension study (ropinirole, n = 70; placebo, n = 77). INTERVENTION: Use of ropinirole or placebo therapy. MAIN OUTCOME MEASURES: The efficacy variables were the number of patients who successfully completed the 12-month study and did not require supplemental levodopa, the number of patients requiring supplemental levodopa, and the proportion of patients having an insufficient therapeutic response. RESULTS: Significantly fewer ropinirole-treated patients met criteria for insufficient therapeutic response (23 [19.8%] of 116) or required the initiation of levodopa therapy (22 [19%] of 116) compared with placebo-treated patients (60 [48%] of 125 patients for insufficient therapeutic response; 57 [45.6%] of 125 patients for additional levodopa). Significantly more ropinirole-treated patients (51 [44.0%] of 116) successfully completed the 12-month study and did not require supplemental levodopa compared with placebo-treated patients (28 [22.4%] of 125). The incidence of adverse experiences and patient withdrawals was low. CONCLUSION: Ropinirole was effective and well tolerated as monotherapy for 12 months in patients with early Parkinson disease. PMID- 9740116 TI - Lacunar infarcts defined by magnetic resonance imaging of 3660 elderly people: the Cardiovascular Health Study. AB - OBJECTIVE: To identify risk factors for and functional consequences of lacunar infarct in elderly people. METHODS: The Cardiovascular Health Study (CHS) is a longitudinal study of people 65 years or older, in which 3660 participants underwent cranial magnetic resonance imaging (MRI). Neuroradiologists read scans in a standard fashion without any clinical information. Lacunes were defined as subcortical areas consistent with infarcts measuring 3 to 20 mm. In cross sectional analyses, clinical correlates were contrasted among groups defined by MRI findings. RESULTS: Of the 3660 subjects who underwent MRI, 2529 (69%) were free of infarcts of any kind and 841 (23%) had 1 or more lacunes without other types present, totaling 1270 lacunes. For most of these 841 subjects, their lacunes were single (66%) and silent (89%), namely without a history of transient ischemic attack or stroke. In multivariate analyses, factors independently associated with lacunes were increased age, diastolic blood pressure, creatinine, and pack-years of smoking (listed in descending order of strength of association; for all, P < .005), as well as maximum internal carotid artery stenosis of more than 50% (odds ratio [OR], 1.81; P < .005), male sex (OR, 0.74; P < .005), and history of diabetes at entrance into the study (OR, 1.33; P < .05). Models for subgroups of single, multiple, silent, and symptomatic lacunes differed only minimally. Those with silent lacunes had more cognitive, upper extremity, and lower extremity dysfunction not recognized as stroke than those whose MRIs were free of infarcts. CONCLUSIONS: In this group of older adults, lacunes defined by MRI are common and associated with factors that likely promote or reflect small vessel disease. Silent lacunes are also associated with neurologic dysfunction. PMID- 9740117 TI - Seven-year survival rate after age 85 years: relation to Alzheimer disease and vascular dementia. AB - OBJECTIVE: To investigate the survival rate in very elderly individuals in relation to Alzheimer disease, vascular dementia, and other mental and physical disorders. DESIGN: A 7-year longitudinal survey. SETTING: Community and institutions in Gothenburg, Sweden. PARTICIPANTS: A representative sample of 494 people aged 85 years. MAIN OUTCOME MEASURES: Results of neuropsychiatric and physical examinations, key informant interview, and computed tomographic scan of the head. Information on mortality was obtained from the parish office. RESULTS: The 7-year survival rate was higher in women (34.5%) than in men (20.3%). Alzheimer disease and vascular dementia predicted 30.7% of deaths in men and 49.7% of deaths in women according to a calculation of population attributable risk (PAR). A regression analysis showed that mortality in men was predicted by the presence of chronic obstructive lung disease (PAR, 18.8), Alzheimer disease (PAR, 16.0), vascular dementia (PAR, 14.7), cancer of the gastrointestinal tract (PAR, 10.2), and skin cancer (PAR, 6.2), and in women by vascular dementia (PAR, 29.4), Alzheimer disease (PAR, 20.3), cerebrovascular disorder (PAR, 12.1), congestive heart failure (PAR, 8.5), hypertension (PAR, 8.0), myocardial infarction (PAR, 6.5), and cancer of the gastrointestinal tract (PAR, 4.3). Life expectancy decreased with severity of dementia, although survival time in individuals with mild Alzheimer disease was not different from that in individuals without dementia. CONCLUSIONS: In extreme old age, Alzheimer disease and vascular dementia influence the mortality rate considerably. However, mild Alzheimer disease does not influence longevity, at least not during the first 7 years. These findings have important public health implications. PMID- 9740118 TI - Sensory modulation of the blink reflex in patients with blepharospasm. AB - OBJECTIVE: To measure the effects of a prepulse stimulus on the blink reflex responses elicited by an electrical stimulation of the supraorbital nerve in patients with blepharospasm with and without an effective sensory trick. DESIGN: Blink reflexes to supraorbital nerve stimulation were preceded in test trials by a prepulse electrical stimulus to the third finger at various leading intervals. SETTING: Ambulatory patients were treated regularly with botulinum toxin in the Neurology Department of the Hospital Clinic in Barcelona, Spain. SUBJECTS: Seventeen patients with dystonic blepharospasm and 11 age-matched control subjects. Eight of the patients with dystonic blepharospasm used a sensory trick to alleviate spasms and 9 did not. MAIN OUTCOME MEASURES: We measured amplitude of R1 and area of R2 responses elicited by the supraorbital electrical stimulus and determined the percentage of facilitation or inhibition induced by the prepulse. RESULTS: Prepulse facilitation occurred in the R1 response at intervals of 60 to 100 milliseconds and was normal in all patients. Prepulse inhibition occurred in the R2 response at intervals between 50 and 200 milliseconds and was abnormally reduced in 11 patients (64.7%), including all 9 patients who did not use a sensory trick and 2 of the 8 patients who did use a sensory trick. There was a positive correlation between absence of sensory trick and abnormality of the prepulse effects (chi(2)= 23.8; P < .001). CONCLUSIONS: Prepulse inhibition of the trigeminofacial reflex is abnormal in a percentage of patients with blepharospasm, and this abnormality occurs more frequently in patients who do not use a sensory trick. This sensory derangement may contribute to the maintenance of the dystonic spasms by reducing the amount of physiological gating from peripheral nerve inputs on trigeminofacial reflexes. PMID- 9740119 TI - Development of Wernicke-Korsakoff syndrome after long intervals following gastrectomy. AB - BACKGROUND: Surgical exclusion of portions of the gastrointestinal tract is a predisposing risk factor for the development of Wernicke-Korsakoff syndrome. When this disease occurs, it is usually within weeks after the gastrointestinal surgery. However, it is not well known that Wernicke-Korsakoff syndrome may occur after a long latent interval following gastrectomy. SETTING: A research-oriented hospital. PATIENTS: Three patients without a history of alcoholism or dietary deprivation developed Wernicke-Korsakoff syndrome 2 to 20 years after undergoing gastrectomy. In these patients, minor changes in dietary habit led to the development of Wernicke-Korsakoff syndrome. CONCLUSIONS: In addition to a long standing latent deficiency in thiamin levels due to defective absorption following gastrectomy or gastrojejunostomy, other minor factors that may influence the intake of thiamin and the need for thiamin in subjects who have undergone gastrectomy may cause a state of thiamin deficiency resulting in Wernicke-Korsakoff syndrome. Results from our study indicate that the following measures are mandatory: educating patients about proper dietary habits, carefully monitoring their thiamin intake, recognizing Wernicke-Korsakoff syndrome early, and treating it immediately with appropriate measures. PMID- 9740121 TI - Tuberous sclerosis. PMID- 9740120 TI - Effectiveness of chloroquine and hydroxychloroquine in treating selected patients with sarcoidosis with neurological involvement. AB - OBJECTIVE: To assess the efficacy of chloroquine (Aralen) phosphate and hydroxychloroquine (Plaquenil) sulfate in the treatment of patients with neurosarcoidosis who either do not respond to corticosteroid therapy or develop unacceptable side effects. DESIGN: Retrospective study. SETTING: Sarcoidosis clinic at a university teaching hospital. PATIENTS: Twelve patients with biopsy proved sarcoidosis, 6 women and 6 men ranging from 20 to 49 years of age, with neurological involvement. INTERVENTION: Chloroquine phosphate, 250 mg twice daily, or hydroxychloroquine sulfate, 200 mg twice daily, was administered for a period of 6 to 21 months. Patients had regular clinical evaluation, determination of serum and cerebrospinal fluid angiotensin-converting enzyme level, computed tomography or magnetic resonance imaging, chest radiography, lung function testing, and slit-lamp examination of the eyes. RESULTS: Chloroquine or hydroxychloroquine either stabilized symptoms or controlled neurological symptoms in 10 of 12 patients. Two patients failed to respond. Cerebrospinal fluid abnormalities, including lymphocytosis, were seen in 3 patients. Magnetic resonance imaging with gadolinium was most helpful in supporting the diagnosis of sarcoidosis and monitoring the course of the disease. CONCLUSIONS: Chloroquine and hydroxychloroquine are effective in controlling neurological sarcoidosis in those patients who fail to respond to corticosteroids or develop severe side effects. Ocular toxic effects from chloroquine or hydroxychloroquine were not observed. PMID- 9740122 TI - The 9/4 secondary structure of eukaryotic selenocysteine tRNA: more pieces of evidence. PMID- 9740123 TI - A 5'-3' exonuclease activity involved in forming the 3' products of histone pre mRNA processing in vitro. AB - Histone RNA 3' processing in vitro produces one or more 5' cleavage products corresponding to the mature histone mRNA 3' end, and a group of 3' cleavage products whose 5' ends are mostly located several nucleotides downstream of the mRNA 3' end. The formation of these 3' products is coupled to the formation of 5' products and dependent on the U7 snRNP and a heat-labile processing factor. These short 3' products therefore are a true and general feature of the processing reaction. Identical 3' products are also formed from a model RNA containing all spacer nucleotides downstream of the mature mRNA 3' end, but no sequences from the mature mRNA. Again, this reaction is dependent on both the U7 snRNP and a heat-labile factor. Unlike the processing with a full-length histone pre-mRNA, this reaction produces only 3' but no 5' fragments. In addition, product formation is inhibited by addition of cap structures at the model RNA 5' end, indicating that product formation occurs by 5'-3' exonucleolytic degradation. This degradation of a model 3' product by a 5'-3' exonuclease suggests a mechanism for the release of the U7 snRNP after processing by shortening the cut off histone spacer sequences base paired to U7 RNA. PMID- 9740124 TI - Circular mRNA can direct translation of extremely long repeating-sequence proteins in vivo. AB - Many proteins with unusual structural properties are comprised of multiple repeating amino acid sequences and are often fractious to expression in recombinant systems. To facilitate recombinant production of such proteins for structural and engineering studies, we have produced circular messenger RNAs with infinite open reading frames. We show that a circular mRNA containing a simple green fluorescent protein (GFP) open reading frame can direct GFP expression in Escherichia coli. A circular mRNA with an infinite GFP open reading frame produces extremely long protein chains, proving that bacterial ribosomes can internally initiate and repeatedly transit a circular mRNA. Only the monomeric forms of GFP produced from circular mRNA are fluorescent. Analysis of the translation initiation region shows that multiple sequences contribute to maximal translation from circular mRNA. This technology provides a unique means of producing a very long repeating-sequence protein, and may open the way for development of proteinaceous materials with novel properties. PMID- 9740125 TI - Differential chemical probing of a group II self-splicing intron identifies bases involved in tertiary interactions and supports an alternative secondary structure model of domain V. AB - Dimethyl sulfate modification was used to probe for tertiary structural elements in the group II intron PI.LSU/2 from the mitochondrial pre-ribosomal RNA of the brown alga Pylaiella littoralis. Modification of the lariat form of the intron under conditions that allow both native folding and conformational homogeneity is found to be generally consistent with secondary and tertiary structural features identified previously for group II ribozymes. A comparison of chemical probing at temperatures just below and above the first melting transition illustrates the cooperative unfolding of tertiary structure and identifies novel candidates for tertiary interactions in addition to defining elements of secondary structure. Substitution of the GAAA terminal loop of domain V is shown to be compatible with retention of conformational homogeneity (despite the loss of an important tertiary interaction), but produces a concise methylation footprint in domain I at the site previously shown to harbor the receptor for that loop. The analysis also identified two nucleotide positions in domain V with novel secondary and potential tertiary structural roles. The proposed refinement of domain V secondary structure is supported by an expanded comparative analysis of group II sequences and bears increased resemblance to U2:U6 snRNA pairing in the spliceosome. PMID- 9740126 TI - Probing of the spliceosome with site-specifically derivatized 5' splice site RNA oligonucleotides. AB - We have developed a site-specific chemical modification technique to incorporate a photoreactive azidophenacyl (APA) group at designated internal positions along the RNA phosphodiester backbone. Using this technique, we have analyzed interactions of the 5' splice site (5'SS) RNA within the spliceosome. Several crosslinked products can be detected within complex B using the derivatized 5'SS RNAs, including U6 snRNA, hPrp8p, and 114-, 90-, 70-, 54-, and 27-kDa proteins. The 5'SS RNAs derivatized at intron positions +4 to +8 crosslink to U6 snRNA, confirming the previously reported pairing interaction between these sequences. hPrp8p and p70 are crosslinked to the 5'SS RNA when the APA is placed within the 5' exon. Finally, a set of unidentified proteins, including p114, p54, and p27, is detected with the 5'SS RNA derivatized at intron positions +4 to +8. Introduction of the bulky APA group near the 5'SS junction (positions -2 to +3) strongly interferes with complex B formation and thus no APA crosslinks are observed at these positions. Together with our earlier observation that hPrp8p crosslinks to the GU dinucleotide at the 5' end of the intron, these results suggest that the inhibitory effect of APA results from steric hindrance of the hPrp8p:5'SS interaction. Unexpectedly, thio-modifications within the region of the 5'SS RNA that is involved in base pairing to U6 snRNA strongly stimulate complex B formation. PMID- 9740127 TI - Specific site selection in RNA resulting from a combination of nonspecific secondary structure and -CCR- boxes: initiation of minus strand synthesis by turnip yellow mosaic virus RNA-dependent RNA polymerase. AB - A turnip yellow mosaic virus RNA-dependent RNA polymerase activity was used to study the template requirements for in vitro minus strand synthesis, which is initiated specifically opposite the 3'-CCA that terminates the 3'-tRNA-like structure. A deletion survey confirmed earlier results suggesting the absence of minus strand promoter elements upstream of the pseudoknotted acceptor stem and 3' terminus. Reiteration of this 27-nt domain provided two competing initiation sites. By varying the added downstream element, it was shown that the pseudoknotted domain could be functionally replaced by various simple stem/loops, although with some decrease in activity. The addition of varying numbers of consecutive -CCA- triplets to the 3' end of the tRNA-like structure resulted in accurate initiation from each added triplet. A similar spectrum of initiations occurred with an unstructured RNA consisting of 12 consecutive -CCA- triplets and no additional viral sequence. Substitution mutations revealed no influence on minus strand synthesis of the identity of the nucleotide immediately upstream of a -CC- initiation site, but a preference for a purine immediately downstream. The introduction of secondary structure into the linear template showed that the usage of potential -CCR- initiation sites is influenced by nonspecific secondary structure. We conclude that specificity arises from the requirement that a -CCR- sequence be sterically accessible. This mechanism is only applicable to interactions that do not involve RNA unwinding during site selection, but may be used commonly in positive strand RNA virus replication and be applicable to other RNA-protein interactions. PMID- 9740128 TI - Intronic snoRNA biosynthesis in Saccharomyces cerevisiae depends on the lariat debranching enzyme: intron length effects and activity of a precursor snoRNA. AB - The eukaryotic small nucleolar RNAs (snoRNAs) are involved in processing of pre rRNA and modification of rRNA nucleotides. Some snoRNAs are derived from mono- or polycistronic transcription units, whereas others are encoded in introns of protein genes. The present study addresses the role of the RNA lariat-debranching enzyme (Dbr1p) in the synthesis and function of intronic snoRNAs in the yeast Saccharomyces cerevisiae. Intronic snoRNA production was determined to depend on Dbr1p. Accumulation of mature intronic snoRNAs is reduced in a dbr1 mutant; instead, intronic snoRNAs are "trapped" within host intron lariats. Interestingly, the extent of intronic snoRNA accumulation in the form of lariats in dbr1 cells varied among different intronic snoRNAs. Intronic snoRNAs encoded within shorter introns, such as U24 and snR38, accumulate more unprocessed lariat precursors than those encoded within longer introns, e.g., U18 and snR39. This correlation was corroborated by experiments conducted with model intron:U24 snoRNA constructs. These results support a splicing-dependent exonucleolytic pathway for the biosynthesis of intronic snoRNAs. Curiously, U24 in a lariat may be functional in directing methylation of ribosomal RNA. PMID- 9740129 TI - Distinct functions of the closely related tandem RNA-recognition motifs of hnRNP A1. AB - hnRNP A1 regulates alternative splicing by antagonizing SR proteins. It consists of two closely related, tandem RNA-recognition motifs (RRMs), followed by a glycine-rich domain. Analysis of variant proteins with duplications, deletions, or swaps of the RRMs showed that although both RRMs are required for alternative splicing function, each RRM plays distinct roles, and their relative position is important. Surprisingly, RRM2 but not RRM1 could support this function when duplicated, despite their very similar structure. Specific RNA binding and annealing are not sufficient for hnRNP A1 alternative splicing function. These observations, together with phylogenetic and structural data, suggest that the two RRMs are quasi-symmetric but functionally nonequivalent modules that evolved as components of a single bipartite domain. PMID- 9740130 TI - Characterization of an anti-RNA recombinant autoantibody fragment (scFv) isolated from a phage display library and detailed analysis of its binding site on U1 snRNA. AB - This is the first study in which the complex of a monoclonal autoantibody fragment and its target, stem loop II of U1 snRNA, was investigated with enzymatic and chemical probing. A phage display antibody library derived from bone marrow cells of an SLE patient was used for selection of scFvs specific for stem loop II. The scFv specificity was tested by RNA immunoprecipitation and nitrocellulose filter binding competition experiments. Immunofluorescence data and immunoprecipitation of U1 snRNPs containing U1A protein, pointed to an scFv binding site different from the U1A binding site. The scFv binding site on stem loop II was determined by footprinting experiments using RNase A, RNase V1, and hydroxyl radicals. The results show that the binding site covers three sequence elements on the RNA, one on the 5' strand of the stem and two on the 3' strand. Hypersensitivity of three loop nucleotides suggests a conformational change of the RNA upon antibody binding. A three-dimensional representation of stem loop II reveals a juxtapositioning of the three protected regions on one side of the helix, spanning approximately one helical turn. The location of the scFv binding site on stem loop II is in full agreement with the finding that both the U1A protein and the scFv are able to bind stem loop II simultaneously. As a consequence, this recombinant monoclonal anti-U1 snRNA scFv might be very useful in studies on U1 snRNPs and its involvement in cellular processes like splicing. PMID- 9740132 TI - Structure of 5S rRNA within the Escherichia coli ribosome: iodine-induced cleavage patterns of phosphorothioate derivatives. AB - The protection patterns of 5S rRNA in solution, within the ribosomal 50S subunit, 70S ribosomes, and functional complexes, were assessed with the phosphorothioate method. About 20% of the analyzed positions (G9-G107) showed strong assembly defects: A phosphorothioate at one of these positions significantly impaired the incorporation of 5S rRNA into 50S particles. The reverse has also been observed: A phosphorothioate is preferred over a phosphate residue in the assembly process at a few positions. The results further demonstrate that 5S rRNA undergoes conformational changes during the assembly in the central protuberance of the 50S subunit and upon association with the small ribosomal subunit forming a 70S ribosome. In striking contrast, when the 70S ribosomes are once formed, the contact pattern of the 5S rRNA is the same in various functional states such as initiation-like complexes and pre- and posttranslocational states. PMID- 9740131 TI - The 5S rRNA loop E: chemical probing and phylogenetic data versus crystal structure. AB - A significant fraction of the bases in a folded, structured RNA molecule participate in noncanonical base pairing interactions, often in the context of internal loops or multi-helix junction loops. The appearance of each new high resolution RNA structure provides welcome data to guide efforts to understand and predict RNA 3D structure, especially when the RNA in question is a functionally conserved molecule. The recent publication of the crystal structure of the "Loop E" region of bacterial 5S ribosomal RNA is such an event [Correll CC, Freeborn B, Moore PB, Steitz TA, 1997, Cell 91:705-712]. In addition to providing more examples of already established noncanonical base pairs, such as purine-purine sheared pairings, trans-Hoogsteen UA, and GU wobble pairs, the structure provides the first high-resolution views of two new purine-purine pairings and a new GU pairing. The goal of the present analysis is to expand the capabilities of both chemical probing and phylogenetic analysis to predict with greater accuracy the structures of RNA molecules. First, in light of existing chemical probing data, we investigate what lessons could be learned regarding the interpretation of this widely used method of RNA structure probing. Then we analyze the 3D structure with reference to molecular phylogeny data (assuming conservation of function) to discover what alternative base pairings are geometrically compatible with the structure. The comparisons between previous modeling efforts and crystal structures show that the intricate involvements of ions and water molecules in the maintenance of non-Watson-Crick pairs render the process of correctly identifying the interacting sites in such pairs treacherous, except in cases of trans-Hoogsteen A/U or sheared A/G pairs for the adenine N1 site. The phylogenetic analysis identifies A/A, A/C, A/U and C/A, C/C, and C/U pairings isosteric with sheared A/G, as well as A/A and A/C pairings isosteric with both G/U and G/G bifurcated pairings. Thus, each non-Watson-Crick pair could be characterized by a phylogenetic signature of variations between isosteric-like pairings. In addition to the conservative changes, which form a dictionary of pairings isosterically compatible with those observed in the crystal structure, concerted changes involving several base pairs also occur. The latter covariations may indicate transitions between related but distinctive motifs within the loop E of 5S ribosomal RNA. PMID- 9740134 TI - College of American Pathologists Conference XXXI on laboratory monitoring of anticoagulant therapy: introduction. PMID- 9740133 TI - Recombination, RNA evolution, and bifunctional RNA molecules isolated through chimeric SELEX. AB - Exchange of RNA structural domains through recombination can be used to engineer RNAs with novel functions and may have played an important role in the early evolution of life. The degree of function an RNA element retains upon recombination into a new sequence context is a measure of how deleterious or beneficial recombination will be. When we fused pairs of aptamers previously selected to bind coenzyme A, chloramphenicol, or adenosine, the chimerae retained some ability to bind both targets, but with reduced binding activity both in solution and on affinity resins, probably due to misfolding. Complex populations of recombined RNAs gave similar results. Applying dual selection pressure to recombined populations yielded the combinations that were best suited to binding both targets. Most reselected RNAs folded into the active conformation more readily than chimerae built from arbitrarily chosen aptamers, as indicated both by solution Kd measurements and affinity resin binding activity. Deletion/selection experiments confirmed that the sequences required for binding are fully contained within the respective domains and not derived from interaction between the domains, consistent with the modular architecture of their original design. The combinatorial nature of the recombination methods presented here takes advantage of the full sequence diversity of the starting populations and yields large numbers of bifunctional molecules (10(6) to more than 1012). The method can be easily generalized and should be applicable to engineering dual-function RNAs for a wide variety of applications, including catalysis, novel therapeutics, and studies of long-range RNA structure. PMID- 9740136 TI - College of American Pathologists Conference XXXI on laboratory monitoring of anticoagulant therapy: laboratory monitoring of unfractionated heparin therapy. AB - OBJECTIVE: To review the state of the art as reflected in the medical literature and the consensus opinion of recognized experts in the field regarding the laboratory monitoring of unfractionated heparin therapy. DATA SOURCES, EXTRACTION AND SYNTHESIS: The authors made an extensive review of the literature. The draft manuscript was circulated to every participant in the consensus conference prior to the convening of the conference. Extensive discussion concerning all of the issues addressed in the manuscript as well as the resulting recommendations occurred. This information was then used to revise the manuscript into its final form. CONCLUSIONS: The resulting manuscript has 23 specific recommendations regarding preanalytic, analytic, and postanalytic phases of monitoring and testing for complications related to unfractionated heparin therapy. This report contains detailed discussion of these recommendations and includes literature citations that support them. A number of issues for which consensus could not be reached are also discussed. A method is provided to assist laboratories, particularly small laboratories, in providing clinicians with an appropriate therapeutic range for the activated partial thromboplastin time, the most commonly used test in monitoring heparin therapy. PMID- 9740135 TI - College of American Pathologists Conference XXXI on laboratory monitoring of anticoagulant therapy: laboratory monitoring of oral anticoagulant therapy. AB - OBJECTIVE: To review the state of the art of laboratory monitoring of oral anticoagulant therapy, as reflected by the medical literature and the consensus opinion of recognized experts in the field, and to make recommendations for improvement in laboratory monitoring of oral anticoagulant therapy. DATA SOURCES: Review of the medical literature, primarily from the last 10 years, and current laboratory practices by a panel of 8 international experts in the field of oral anticoagulant monitoring. DATA EXTRACTION AND SYNTHESIS: After an initial assessment of the literature, key points were identified. Experts were assigned to do an in-depth review of the literature and current practices relevant to each of the key points and to prepare a summary of their findings and recommendations. A draft manuscript was prepared and circulated to every participant in the College of American Pathologists Conference XXXI on Laboratory Monitoring of Anticoagulant Therapy prior to the conference. Each of the key points and associated recommendations was then presented for discussion at the Conference. Recommendations were accepted if a consensus of the 26 experts attending the Conference was reached. The results of the discussion were used to revise the manuscript into its final form. CONCLUSIONS: Consensus was reached on 12 recommendations concerning the laboratory monitoring of oral anticoagulant therapy. Detailed discussion of the rationale for each of these recommendations is found in the text of this article. Discussion of points on which consensus was not reached is also included in the text. It is hoped that widespread adoption of these recommendations will further improve the laboratory monitoring of oral anticoagulant therapy. PMID- 9740138 TI - Thrombosis: diagnosis and prophylaxis 1997. PMID- 9740137 TI - College of American Pathologists Conference XXXI on laboratory monitoring of anticoagulant therapy: the clinical use and laboratory monitoring of low molecular-weight heparin, danaparoid, hirudin and related compounds, and argatroban. AB - OBJECTIVE: To review the role of the laboratory in monitoring therapy with low molecular-weight heparin, danaparoid, hirudin, and argatroban, as reflected in the medical literature and the consensus opinion of recognized experts in the field. DATA SOURCES: Review of the medical literature and current clinical practice by a panel of 6 international experts in the field of anticoagulant therapy. DATA EXTRACTION AND SYNTHESIS: The experts made an extensive review of the published literature and prepared a draft manuscript, which included preliminary recommendations. The draft manuscript was circulated to participants in the College of American Pathologists Conference XXXI on Laboratory Monitoring of Anticoagulant Therapy prior to the conference. The manuscript and recommendations were then presented at the Conference for discussion. Recommendations were accepted if a consensus of the 26 experts attending the Conference was reached. The results of the discussion were used to revise the manuscript into its final form. CONCLUSIONS: This report reviews the mechanism of action and potential uses of these newer anticoagulant agents. General guidelines for monitoring these agents and 9 specific recommendations for laboratory monitoring of low-molecular-weight heparin and danaparoid are provided, along with citation of the appropriate supporting literature. Issues for which a consensus was not reached at the Conference are also discussed. PMID- 9740139 TI - Glycoprotein IIb-IIIa in platelet aggregation: an emerging target for the prevention of acute coronary thrombotic occlusions. PMID- 9740140 TI - The new anticoagulants: new opportunities, new issues. PMID- 9740141 TI - A clinician's view of thrombotic disorders of the cardiovascular system. PMID- 9740142 TI - Leukocyte infiltration and intercellular adhesion molecule-1-mediated cell interactions in immunoglobulin A nephropathy. AB - BACKGROUND: Mononuclear leukocytes have a role in immunoglobulin (Ig) A nephropathy. Renal leukocyte recruitment is mediated by adhesive interactions between leukocytes and their ligands on renal cells. METHODS: We assessed interstitial and glomerular leukocytes using an avidin-biotin-peroxidase method with monoclonal antibodies (MAbs) against leukocytes (CD45), beta2-integrin (CD18), macrophages (CD14), T cells (CD3) and T-cell subsets (CD4, CD8), intercellular adhesion molecule-1 (ICAM-1) (CD54), and HLA class II antigens. We analyzed their relation with the abnormal expression of ICAM-1 on proximal tubular epithelium in sequential renal sections from 57 patients with IgA nephropathy stratified by degrees of interstitial cellular infiltration as seen by light microscopy. RESULTS: In IgA nephropathy without infiltration (n = 15) and with interstitial cellular infiltration of 1+ by light microscopy (n = 11), ICAM-1 expression on vascular endothelium was unchanged with respect to that observed in the normal kidney; the proximal tubular epithelium was negative for this stain. Moreover, in IgA nephropathy with interstitial cellular infiltration greater than or equal to 2+ by light microscopy (n = 31), ICAM-1+ stain was seen on proximal tubular epithelium. The value of ICAM-1+ tubule staining was significantly (P = .0004) higher in the biopsies with a higher degree of interstitial cellular infiltration seen by light microscopy. The tubular ICAM-1+ stain was significantly associated with the number of interstitial leukocytes identified by the MAbs tested and correlated (P < .05) with CD45+ (r = 0.59), CD14+ (r = 0.54), and CD3+ (r = 0.51) interstitial leukocytes in IgA nephropathy with interstitial cellular infiltration by light microscopy. Interstitial ICAM-1+ cells and interstitial CD18+ leukocytes were correlated (r = 0.85, P < .001). Correlation was found between the value of ICAM-1+ tubule staining and the number of CD45+ (r = 0.87, P < .01), CD14+ (0.58, P < .05), and CD8+ (r = 0.68, P < .05) glomerular leukocytes. CONCLUSIONS: Our results suggest that tubular and interstitial ICAM-1+ cells may participate in adhesive interactions with interstitial leukocytes. Epithelial tubular cells expressing ICAM-1 may be a marker of tubulointerstitial disturbance upregulated by interstitial and glomerular infiltration of macrophages and T cells in IgA nephropathy. PMID- 9740143 TI - The utility of parotid gland and level I and II neck fine-needle aspiration. AB - OBJECTIVE: Although fine-needle aspiration (FNA) commonly is used in the diagnostic workup of parotid gland and level I and II neck lesions, the effect of an FNA service on patient care has not been definitively established. METHODS: Follow-up was obtained in 158 patients who underwent FNA. The value of FNA was analyzed by determining the proportions of cases in which management was altered by the information obtained. RESULT: The percentage of lesions classified by FNA as benign, nonneoplastic; benign, neoplastic; atypical or suspicious; malignant; and insufficient was 42%, 28%, 16%, 41%, and 7%, respectively. By using FNA, an operation was avoided in 70% and 79% of patients with a nonneoplastic lesion and a metastasis, respectively. CONCLUSIONS: Although definitive subclassification of some lesion types was poor, FNA was useful in patient triage. PMID- 9740145 TI - Needle biopsies of the prostate: what constitutes adequate histologic sampling? AB - OBJECTIVE: The automated biopsy gun and increased screening for adenocarcinoma of the prostate have led to increased numbers of biopsies with only tiny foci of prostatic carcinoma. Consequently, the risk of failing to sample a small focus of carcinoma histologically has increased as well. Most pathologists routinely sample prostatic needle biopsies at more than 1 level. An expert panel has recently suggested that prostatic needle biopsies be sampled at at least 2 levels. However, there have been no studies measuring the amount of additional tissue sampled by multiple levels versus 1 level. METHODS: Forty-two prostatic needle biopsies were serially sectioned at 4-microm levels. Hematoxylin-eosin stained slides were prepared from every fifth section. The total length of each biopsy was compared with the length sampled by 1 level (50% through the block) and 3 levels (25%, 50%, and 75% through the block). RESULTS: Sampling the tissue at 1 level missed an average of 23.4% of the total biopsy length. Sampling the tissue at 3 levels significantly improved this average to 7% (P = .0001). CONCLUSIONS: This study shows that a single histologic section of a prostatic needle biopsy often fails to sample a significant portion of available tissue. This could occasionally result in failure to sample a small focus of prostatic carcinoma. The authors recommend that prostatic needle biopsies be routinely sampled at 3 levels (approximately 25%, 50%, and 75% through the block). PMID- 9740144 TI - High expression of growth factors and growth factor receptors in ovarian metastases from ileal carcinoids: an immunohistochemical study of 2 cases. AB - OBJECTIVE AND DESIGN: Ovarian metastatic carcinoids are rare neoplasms that show prominent fibrosis of tumor stroma and are often associated with peritoneal carcinomatosis. We studied formalin-fixed and paraffin-embedded tumor specimens of 2 cases of ovarian metastases from ileal enterochromaffin cell carcinoids immunohistochemically to evaluate whether acidic fibroblast growth factor (aFGF), transforming growth factor-alpha (TGFalpha), and their respective receptors (fibroblast growth factor receptor-4 [FGFR4] and epidermal growth factor receptor [EGFR]) may play a role in the pathogenesis of stromal fibroblast reaction and in the mechanism of tumor dissemination. RESULTS: In both cases, the majority of tumor cells expressed immunoreactivity for aFGF, FGFR4, and TGFalpha. Immunoreactivity for FGFR4 was detected in stromal cells of both cases, while EGFR-positive stromal cells were found in only 1 case. Immunoreactivity for FGFR4 was also found in peritoneal mesothelial cells. CONCLUSIONS: The coexpression of aFGF and FGFR4 in neoplastic enterochromaffin cells suggests that aFGF may act as an autocrine factor stimulating tumor cell growth. In addition, aFGF and TGFalpha may stimulate, in a paracrine fashion, the proliferation of FGFR4- and EGFR immunoreactive stromal fibroblasts. Finally, interaction of aFGF-immunoreactive enterochromaffin cells with FGFR4-bearing mesothelial cells may play a role in the mechanism of serosal implant and spread of tumor cells. PMID- 9740146 TI - MaMi, a human endometrial stromal sarcoma cell line that constitutively produces interleukin-6, interleukin-8, and monocyte chemoattractant protein-1. AB - BACKGROUND: Uterine endometrial stromal sarcoma is a rare neoplasm with a morphology that closely resembles that of the proliferative endometrial stroma. To understand its pathologic characteristics, we established a novel cell line, MaMi, from a primary culture of an endometrial stromal sarcoma obtained from a 65 year-old Japanese woman. METHODS: We observed the morphology of MaMi cells and performed immunohistochemical analysis on the primary tumor and transplants in nude mice. Prolactin, insulin-like growth factor-binding protein-1, interleukin (IL)-6, IL-8, monocyte chemoattractant protein-1, intercellular adhesion molecule 1, E-selectin, vascular cell adhesion molecule-1, and fibronectin production in the culture medium of MaMi cells were also examined. RESULTS: MaMi cells were shown to exhibit a fibroblast-like morphology in vitro, and they adopted a more elongated appearance in response to 12-O-tetradecanoyl phorbol-13-acetate (TPA). On injection into nude mice, the cells gave rise to subcutaneous tumors. Immunohistologically, both the primary tumor and MaMi cell-induced tumors stained positively with antibodies to neuron-specific enolase or vimentin. MaMi cells constitutively produced IL-6, IL-8, and monocyte chemoattractant protein-1 in vitro. Interleukin-1beta, (100 pmol/L), tumor necrosis factor-alpha (1 nmol/L), and lipopolysaccharide (1 microg/mL) each increased the release of IL-6, IL-8, and monocyte chemoattractant protein-1 by MaMi cells. TPA (10 nmol/L) also stimulated the production of IL-6 and IL-8 by these cells, but inhibited that of monocyte chemoattractant protein-1. CONCLUSIONS: We demonstrated that MaMi cells closely resemble proliferative endometrial stromal cells not only morphologically, but also functionally. This cell line may prove valuable in understanding the role of cytokines produced by tumor cells in the pathogenesis of endometrial stromal sarcoma and may also be useful as an in vitro model of functioning endometrial stromal cells. PMID- 9740147 TI - Uterine arteriovenous malformation necessitating hysterectomy with bilateral salpingo-oophorectomy in a young pregnant patient. AB - This case report describes a 31-year-old woman at 8 weeks' gestation with large arteriovenous malformation of the uterus involving bilateral uterine and ovarian arteries. She had a history of multiple pregnancy losses, as well as spontaneous copious vaginal hemorrhage. The patient underwent an embolization procedure followed by total abdominal hysterectomy and bilateral salpingo-oophorectomy. The uterus was very small (30 g) despite its gravid status, and the overall microscopic findings indicated Mullerian system hypoplasia in addition to vascular malformation. PMID- 9740148 TI - Epithelioid hemangioendothelioma of the liver mimicking Budd-Chiari syndrome. AB - A case of epithelioid hemangioendothelioma of the liver in a 34-year-old man with clinical and radiologic findings suggestive of Budd-Chiari syndrome is reported. Despite clinical and radiologic findings, percutaneous liver biopsy was suspicious for epithelioid hemangioendothelioma. The patient underwent liver transplantation 2 months later, and histologic examination confirmed this diagnosis. Unusual histopathologic features included extensive areas of capillary thin vascular structures with open lumina, lack of significant cytologic atypia in the majority of neoplastic cells, and areas with Budd-Chiari-like features in the hepatic parenchyma surrounding the tumor. The neoplastic cells were focally immunopositive for endothelial markers, such as factor VIII-related antigen and CD34 antigen. The unusual clinical presentation may have been due to tumor invasion and fibrous obliteration of terminal hepatic venules and sublobular veins. Epithelioid hemangioendothelioma should be considered when evaluating patients with clinical features of Budd-Chiari syndrome or veno-occlusive disease. PMID- 9740149 TI - Chinese herbs nephropathy or the evils of nature. PMID- 9740150 TI - Analgesic nephropathy. AB - Many questions about analgesic nephropathy (AN) lack clear-cut answers. We present available evidence for and against proposed answers to many of these questions. These include: (1) Is acetaminophen (AC) nephrotoxic when taken as the sole analgesic? (2) Is the combination of acetylsalicylic acid (ASA) and AC more nephrotoxic than AC taken alone, and if so, why? (3) What are the minimum doses and durations of ingestion required to produce analgesic nephrotoxicity? (4) Is the combination of ASA and AC (a major metabolite of phenacetin) less nephrotoxic than that of phenacetin and ASA combined? (5) Does caffeine in combination with analgesics contribute to nephrotoxicity? (6) What is the incidence of end-stage renal disease (ESRD) due to AN? (7) What uniform diagnostic criteria should be established for AN? (8) What are the earliest anatomic and biochemical abnormalities? (9) What are the mechanisms of renal injury? (10) Does AC cause uroepithelial neoplasia? (11) What research might be most beneficial? Based mainly on associations, some strong, we suggest that AN still exists as a cause of ESRD in the United States, where AC/ASA combinations are available over the counter, and in Canada, where they are not. We also suggest that the evidence needed to recommend that the AC/ASA combination be excluded from over-the-counter analgesic preparations still has limitations. A prospective multicenter study comparing incidence related to AC/ASA in the United States and to AC in Canada and the United States may be needed to answer this question. For such a study to be worthwhile, an adequate incidence in both countries is required. PMID- 9740151 TI - Falciparum malaria and the kidney: a model of inflammation. AB - Renal and renal-related disorders commonly occur in infection with Plasmodium falciparum, which can cause fluid and electrolyte disorders, glomerulonephritis, and acute renal failure (ARF). It appears that ARF and other life-threatening complications in falciparum malaria are not directly caused by the parasite itself but are the result of interaction of mechanical, immunologic, and humoral components. P. falciparum-infected erythrocytes impair microcirculation and cause hemolysis. Glycosylphosphatidylinositol moieties covalently linked to the surface antigens of falciparum malarial parasites appear to act like endotoxin. Glycosylphosphatidylinositol, via CD14, which is a receptor on monocytes, stimulates monocytes to release tumor necrosis factor, which in turn enhances synthesis of various cytokine cascades and mediators. Besides contributing to ARF, these mediators also cause changes in blood volume status. The degree of vasodilatation caused by vasodilating mediators varies with the severity of infection. Increased vascular permeability by the mediators occurs in severe infection, which results in hypovolemia and contributes to ARF. Although the cornerstone of treatment of malaria still is antimalarial drugs, several new modalities of treatment targeting toxin, signal transduction, mediators, and cytokines have great potential. PMID- 9740152 TI - A case control study of proximal calciphylaxis. AB - The purpose of this investigation was to describe the clinical presentation of nine patients with calciphylaxis involving the proximal lower extremities or trunk and to compare the clinical characteristics of these patients with those of 347 hemodialysis patients from the same geographic area. Patients were identified primarily through a computer search of pathology records, identifying patients with the term "calciphylaxis" in the biopsy report. All patients had pathologic specimens consistent with calciphylaxis. All the calciphylaxis patients were white and were markedly obese. While two patients had markedly elevated parathyroid hormone levels, most patients did not show severe derangements of calcium phosphate metabolism compared with other dialysis patients. A logistic regression model identified body mass index and low serum albumin 3 months before diagnosis as being highly associated with a diagnosis of calciphylaxis. Diabetes mellitus and parameters of calcium-phosphate metabolism were not significantly associated with proximal calciphylaxis. These findings suggest that white race, morbid obesity, and poor nutritional status are associated with proximal calciphylaxis in dialysis patients. PMID- 9740153 TI - Cutaneous necrosis from calcific uremic arteriolopathy. AB - Calcific uremic arteriolopathy (calciphylaxis) is an uncommon complication of chronic renal failure that is associated with high morbidity and mortality. We report 16 patients (13 female) who presented between 1985 and 1996. All patients developed painful livido reticularis that progressed to cutaneous necrosis and ulceration (11 cases on the proximal extremities and five cases on the distal extremities). Two patients with predominately distal leg disease survived; the cause of death in the other 14 patients was sepsis (six patients), withdrawal from dialysis (three), cardiac arrest (three), and gastrointestinal hemorrhage (two). Mesenteric ischemia from intestinal vascular calcification occurred in two cases. Clinical factors identified included the use of warfarin therapy in seven cases and significant weight loss (>10% body weight) in seven cases in the 6 months preceding the development of calcific uremic arteriolopathy. Skin pathology was studied in 12 cases, with all showing calcific panniculitis and small vessel calcification. Electron microscopic spectral analysis of the mineral content of the calcific lesions in the subcutaneous tissue showed only calcium and phosphorous. In two cases, substitution of low molecular weight heparin for warfarin therapy resulted in clinical improvement. Current theories of pathogenesis and treatment are reviewed. This study confirms the high morbidity and mortality of calcific uremic arteriolopathy producing ischemic tissue necrosis while drawing attention to significant weight loss and warfarin therapy as risk factors for the development of ischemic tissue necrosis. Hyperbaric oxygen therapy warrants further study. PMID- 9740155 TI - Cefazolin in chronic hemodialysis patients: a safe, effective alternative to vancomycin. AB - Vancomycin use is common in hemodialysis patients, due in part to the ease of dosing, but can lead to the development of resistant organisms, including vancomycin-resistant enterococcus. Alternate antibiotics may be equally effective and allow similar dosing in the chronic hemodialysis population. A retrospective review of culture results from a 217-patient, non-hospital-based outpatient hemodialysis center was performed over a 7-month period. Wound and blood culture sensitivity to cefazolin, vancomycin, cefazolin plus gentamicin, and vancomycin plus gentamicin was analyzed. Cefazolin was equivalent to vancomycin for empiric treatment of clinically significant infections in a population with a low rate of methicillin-resistant Staphylococcus aureus infection. Cefazolin plus gentamicin was superior to vancomycin alone. The vancomycin plus gentamicin combination did provide minimally broader coverage than the cefazolin plus gentamicin combination. A prospective pharmacokinetic analysis of postdialysis cefazolin dosing was performed in anuric chronic hemodialysis patients dialyzed with polysulfone dialyzers. Peak, predialysis, and postdialysis cefazolin levels were obtained. Nondialysis clearance of cefazolin was sufficiently low (k(e), 0.027; t(1/2), 26.4 hours) and dialysis clearance sufficiently high (k(e), 0.254; t(1/2), 3.19 hours) to provide for safe and effective peak and trough cefazolin levels with postdialysis dosing in anuric hemodialysis patients. In conclusion, cefazolin alone or with gentamicin in an appropriate empiric antibiotic choice in chronic hemodialysis patients dialyzed in a nonhospital setting with low methicillin-resistant S. aureus infection rates. For infections with documented sensitivity to cefazolin, a 1 g intravenous dose postdialysis (750 mg in patients weighing <50 kg) is safe and effective. PMID- 9740154 TI - Identification of crystals in kidneys of AIDS patients treated with foscarnet. AB - Three acquired immune deficiency syndrome patients given foscarnet to treat cytomegalovirus retinitis developed renal failure with crystal deposits within the renal glomeruli. We identified these crystals as a mixture of sodium salt, calcium salt, and a mixed salt containing both sodium and calcium ions. This composition has not been previously reported. Foscarnet can complex available ionized calcium and secondarily precipitate in glomeruli. The percentage of complexing depends on calcium concentration in serum and the poor calcium salt solubility. PMID- 9740156 TI - Cefazolin as empiric therapy in hemodialysis-related infections: efficacy and blood concentrations. AB - Concern about the increasing incidence of vancomycin-resistant organisms has tempered the enthusiasm for indiscriminate vancomycin use. Cefazolin has an antibacterial activity profile similar to vancomycin against most pathogens encountered in the hemodialysis (HD) population. We evaluated the clinical efficacy and serum concentrations that were achieved during empiric cefazolin use. Fifteen consecutive HD patients (five, conventional HD; five, high efficiency HD; and five, high-flux HD) with suspected or documented infections warranting antibiotic intervention, including access-related, respiratory tract, urinary tract, or wound infections, were enrolled. Each patient received intravenous cefazolin (20 mg/kg actual body weight rounded to the nearest 500-mg increment [range, 1 to 2 g]) after each dialysis treatment for at least three doses. Cefazolin concentrations were obtained before and immediately after the next three consecutive dialysis treatments. Thirteen patients were evaluated for efficacy and all 15 were evaluated for toxicity and cefazolin blood concentrations. All patients showed at least a short-term (3-week) clinical resolution of infection with cefazolin treatment. No central nervous system toxicities were noted and no other adverse events were expressed by the patients during the course of cefazolin treatment. Predialysis cefazolin concentrations, as determined by high-performance liquid chromatography, were 70.2 +/- 42.7 (conventional HD), 45.6 +/- 18.9 (high-efficiency HD), and 41.6 +/- 23.9 mg/L (high-flux HD) over the three dialysis sessions. Cefazolin at doses of approximately 20 mg/kg administered post-HD appears to be a safe and effective empiric therapy and yields predialysis cefazolin concentrations of 2.5 times or greater than those considered to be the minimum inhibitory concentration breakpoint (16 mg/L) for susceptible organisms. These data support the broader use of cefazolin for empiric treatment in the HD population, allowing vancomycin to be reserved for confirmed resistant organisms. PMID- 9740157 TI - Vancomycin-resistant enterococcus in end-stage renal disease. AB - The percentage of nosocomial vancomycin-resistant enterococci (VRE) has been increasing rapidly in the United States. This has recently resulted in recommendations to reserve vancomycin use for cases with proven resistance to other antimicrobials. We prospectively investigated the incidence of VRE in our dialysis population and compared it with a control group of 40 clinic patients with chronic renal insufficiency (CRI) who had a serum creatinine level greater than 1.5 mg/dL, but were not undergoing dialysis. The incidence of VRE on our campus is almost 10%, which is similar to US data. We studied 50 chronic hemodialysis (HD) patients and 50 peritoneal dialysis (PD) patients. Each patient had a rectal swab test performed and cultured for the presence of enterococci. Antimicrobial exposures over the 6 months before the initial swab test were reviewed in each patient. At least one repeated swab test was performed in 30 CRI, 45 HD, and 37 PD patients. From the initial swab culture, vancomycin sensitive enterococci (VSE) were isolated in 65% of CRI, 54% of HD, and 70% of PD patients. No CRI or HD patients had VRE isolated and 2% (1 of 50) of PD patients had VRE isolated. The remaining patients had no enterococci isolated. Review of antimicrobial exposures in the 6 months before the initial swab test showed 0% of CRI, 32% of HD, and 36% of PD patients received vancomycin. Other antimicrobials were administered to 40% of CRI, 46% of HD, and 78% of PD patients in the same time period. In the month immediately preceding the initial swab test, 0% of CRI, 12% of HD, and 22% of PD patients received vancomycin and 18% of CRI, 20% of HD, and 36% of PD patients received other antimicrobials. Results from repeated cultures showed that 57% of CRI, 40% of HD, and 38% of PD patients changed their culture status related to VSE, VRE, or no enterococci present. Cultures of 342 swabs from 140 patients yielded three VRE isolates in two patients. We conclude that despite the frequent use of vancomycin and other antimicrobials, the incidence of VRE in our renal population is less than the reported incidence. Given this lack of VRE isolates, we recommend the continued judicious use of vancomycin in treating renal patients and continued enterococcal sensitivity surveillance. PMID- 9740158 TI - Relevance of periglomerular myofibroblasts in progression of human glomerulonephritis. AB - To clarify the pathological and clinical significance of periglomerular alpha smooth muscle actin (alpha-SMA)-positive cells, we examined 51 needle-biopsy specimens from patients with human glomerulonephritis. Immunoelectron microscopy confirmed these cells were myofibroblasts showing characteristic features with abundant alpha-SMA-positive thin myofilaments. Nonsclerotic glomeruli with periglomerular myofibroblasts were larger in the Bowman's capsular planar area than nonsclerotic glomeruli without periglomerular myofibroblasts (24.7 +/- 6.0 x 10(3) microm2 v 19.9 +/- 8.5 x 10(3) microm2; P < 0.01). We studied the correlation between the clinical prognosis and the extent of periglomerular myofibroblasts in 24 patients with IgA nephropathy. Patients were divided into two groups; those with plasma creatinine levels within normal range at biopsy and significantly elevated at follow-up were designated group 1 (poor prognosis), and patients with plasma creatinine levels within normal range at biopsy and not significantly elevated at follow-up were designated group 2 (fair prognosis). In the kidneys of group 1 patients, periglomerular alpha-SMA was expressed more intensively than it was in the kidneys of group 2 patients (alpha-SMA expression score, 1.0 +/- 0.48 v 0.52 +/- 0.54; P < 0.05). These findings indicate that periglomerular myofibroblasts appeared surrounding the nonsclerotic hypertrophic glomeruli, which may lead finally to glomerulosclerosis. This report suggests that interaction between the glomerular cells and the periglomerular myofibroblasts may have a role in the progression of glomerular diseases. PMID- 9740159 TI - A randomized, prospective, comparative study of manual and automated renal biopsies. AB - A percutaneous renal biopsy can be performed in several ways, including using a spring-loaded biopsy gun. As this form of renal biopsy has become more popular, a controversy has developed regarding tissue adequacy and the incidence of complications. To compare these two aspects in an automated biopsy and a manual biopsy, we studied 166 patients assigned to one of the two renal biopsy methods. In a randomized, prospective manner from June 1994 until February 1997, group 1 (67 patients) received a 14 G Tru-cut needle (Baxter, Deerfield, IL) manual biopsy while group 2 (99 patients) received an 18 G automated gun biopsy. There was no difference in sex, age, hemoglobin level, prothrombin time, partial thromboplastin time, or diastolic and systolic blood pressure prebiopsy in groups I and II. Indications for biopsy were proteinuria (38%), proteinuria accompanied by hematuria (31.3%), acute renal failure (9.6%), lupus nephropathy (9.6%), chronic renal failure (6%), and hematuria only (5.4%). In group I, the number of cores was 1.88 +/- 0.56, the glomeruli obtained were 27.3 +/- 13.8, and the number of glomeruli per core were 15.3 +/- 8.4. In group II, the values were 2.37 +/- 0.88, 20.7 +/- 11.1, and 9.95 +/- 6.9, respectively. These results showed a statistically significant difference (P < 0.05). In all cases, pathological diagnosis was possible. The histology showed IgA nephropathy in 25.9%, minimal change disease in 16.3%, lupus nephritis in 11.4%, membranous glomerulonephropathy in 9.3%, membranoproliferative glomerulonephritis in 5.4%, and others. The incidence of postbiopsy hematoma was marginally greater in group I (22.3% v 11.1%) and the area of perirenal hematoma shown on ultrasound 24 hours postbiopsy was larger in group I, as well (848 +/- 623 mm2 v 338 +/- 260 mm2). Hematocrit levels before and after biopsy showed a significant difference (34.9% +/- 7.9% and 34.0% +/- 7.6%, respectively; P < 0.05) in group I, but no significant difference was observed in group II (35.1% +/- 7.0% and 34.7% +/- 6.9%). Both techniques rendered adequate tissue sampling, but the extent of bleeding was more severe with the manual 14 G Tru-cut needle biopsy. PMID- 9740160 TI - Prognosis of patients with acute renal failure requiring dialysis: results of a multicenter study. AB - Despite several decades of clinical experience, the mortality rate for patients with acute renal failure (ARF) requiring dialysis remains high, and the evaluation of the patients prognosis has been difficult. To date, the Acute Physiology and Chronic Health Evaluation II (APACHE II) scoring system has been used more frequently for prediction in studies of ARF than any other scoring system, but has not been prospectively validated in controlled multicenter studies of this entity. In a multicenter, prospective, controlled trial evaluating the use of biocompatible hemodialysis membranes (BCMs) in patients with ARF, we evaluated the extent to which the APACHE II scoring system, based on the physiological variables in the 24 hours before the onset of dialysis and the presence or absence of oliguria, is predictive of outcome. Analysis of survival and recovery of renal function for the 153 patients treated in this study show that APACHE II scores are predictive both of survival and recovery of renal function, whether analyzed separately by type of dialysis membrane used (BCM or bioincompatible [BICM]) or for both groups combined (all P < 0.01). There was no evidence of a significant center effect or interaction of APACHE II score with dialysis membrane in our study. After adjusting for the APACHE II score, there was a positive effect of the BCM on both probability of survival (P < 0.05) and recovery of renal function (P < 0.01). In patients dialyzed with BCMs, oliguria at onset of dialysis had an adverse effect on both survival and recovery of renal function (both P < 0.01). Receiver operator curves (ROCs) using APACHE II score and the use of BCMs in nonoliguric patients yielded a statistically significant improvement versus the use of APACHE II score alone in the area under the curve (AUC) for survival (0.747 to 0.801; P < 0.05) and recovery of renal function (0.712 to 0.775; P < 0.05). We conclude that the use of the APACHE II score determined at the time of initiation of dialysis for patients with ARF is a statistically significant predictor of patient survival and recovery of renal function. The use of the APACHE II score measured at the time of dialysis initiation, especially when modified by the presence or absence of oliguria, should help in predicting outcome when evaluating interventions for patients with ARF. PMID- 9740161 TI - Catabolism in critical illness: estimation from urea nitrogen appearance and creatinine production during continuous renal replacement therapy. AB - Thirty-eight intensive care unit (ICU) patients (26 men and 12 women with a mean age of 57.0 +/- 16.6 years) with acute renal failure (ARF) treated by venovenous continuous renal replacement therapy (CRRT) were evaluated while in relatively steady metabolic control. Twenty-seven were undergoing continuous venovenous hemodialysis, nine were undergoing continuous venovenous hemodiafiltration, and two were undergoing continuous venovenous hemofiltration. Periods of analysis varied between 24 and 408 hours (mean duration, 82.7 +/- 70.6 hours; median, 72 hours). Their mean Acute Physiology and Chronic Health Evaluation II (APACHE II) score within 24 hours of admission to the ICU was 21.3 +/- 6.3 and survival rate was 31.6%. Urea nitrogen and creatinine concentrations were determined every 6 to 12 hours in both serum (Cun and Cc, respectively) and effluent (spent dialysate and/or ultrafiltrate). The mean effluent rate was 1,472 +/- 580 mL/h and blood flow rate, 166 +/- 32 mL/min. Urine was collected daily for urea nitrogen and creatinine measurement. Urea nitrogen appearance rate (UnA) and creatinine production rate (Pc), calculated from urea nitrogen (UnMR) and creatinine mass removal (CMR) from both the effluent and the urine, using Garred mass balance equations and the Forbes-Bruining formula, allowed normalized protein catabolic rate (nPCR) and estimates of lean body mass (LBM) to be derived. Creatinine metabolic degradation rate (Dc), estimated by the Mitch formula, was included in the calculation. The lowest body weight recorded during the study period was considered as dry weight (BW). The creatinine index (CI) was also obtained. For each parameter, the results are presented as mean, median, and range values: UnMRe (from effluent), 13.6 +/- 7.2, 12.5, 1.6 to 32.6 mg/min; UnMRu (from urine), 0.13 +/- 0.40, 0, 0 to 2.30 mg/min; UnA, 13.6 +/- 7.0, 12.5, 3.8 to 32.1 mg/min; nPCR, 1.75 +/- 0.82, 1.60, 0.61 to 4.23 g/kg/d; CMRe (from effluent), 942.0 +/- 362.3, 918.0, 211.2 to 1,641.6 mg/d; CMRu (from urine), 44.4 +/- 138.8, 0, 0 to 698.5 mg/d; Dc, 94.6 +/- 49.9, 81.9, 31.0 to 294.1 mg/d; Pc total, 1,067.1 +/- 409.7, 1,053.7, 261.5 to 1,988.2 mg/d; LBM, 38.3 +/- 11.9, 37.9, 15.0 to 65.0 kg; LBM/BW ratio, 49.5% +/- 14.0%, 50.3%, 22.5% to 86.0%; and CI, 13.7 +/ 4.7, 14.2, 4.1 to 25.8 mg/kg/d. When Pc was estimated from the Cockcroft-Gault equations (as Pc'), the mean value for Pc and Pc' was similar (1,067.1 +/- 409.7 v 1,284.9 +/- 484.1 mg/d), but there were relatively large differences for the majority of cases. A positive correlation was observed between UnA and Pc (R = 0.42). Serum albumin and LBM/BW correlated poorly (R = 0.20). Outcome was weakly related to UnA and to nPCR (R = 0.29 and R = 0.31, respectively). Urea nitrogen appearance appears widely variable in critically ill ARF patients. This simple approach can provide useful information for an easy estimate of net protein catabolism in critically ill patients with ARF undergoing CRRT. PMID- 9740162 TI - Recombinant human growth hormone therapy in malnourished dialysis patients: a randomized controlled study. AB - Recombinant human growth hormone (rhGH; Saizen, Serono, Spain) has been recently used as an anabolic agent in several catabolic states, including malnourished chronic dialysis patients. However, up-to-date, comparative studies with control groups of dialysis patients have not been reported. The aim of the present study was to assess the effects of rhGH on nutritional status in a group of malnourished adult chronic dialysis patients undergoing both continuous ambulatory peritoneal dialysis (CAPD) and hemodialysis (HD). The patients were randomly assigned to the control group (nine patients; 6 women, 3 men; mean age, 58.3 +/- 5.6 years; seven undergoing CAPD, two undergoing HD) or the rhGH group (eight patients; three women, five men; mean age, 63.9 +/- 3.1 years; four undergoing CAPD, four undergoing HD). Both groups were similar at baseline. All patients were given dietary prescriptions (35 kcal/kg/d and 1 g protein/kg ideal body weight/d) during 4 weeks. In the rhGH group, rhGH was administered at 0.2 IU/kg/d subcutaneously (SC) during this period. Anthropometric and analytic parameters were assessed before (0 weeks) therapy and at 2 and 4 weeks after starting therapy. The rhGH group showed an increase of 1.238 kg in body weight from 64.3 +/- 4.3 (mean +/- standard error of the mean [SEM]) to 65.6 +/- 4.9 kg (P < 0.05). Serum insulin-like growth factor type 1 (IGF-1) concentrations increased from 216.6 +/- 42.5 to 581.2 +/- 171.5 ng/mL (4 weeks; P < 0.01) and transferrin levels increased from 271.2 +/- 16.3 to 314.5 +/- 21.2 mg/dL (4 weeks; P < 0.05). A significant reduction in blood urea nitrogen (BUN) level was observed (62.1 +/- 1.8 v 46.8 +/- 3.8 mg/dL; 4 weeks; P < 0.05). Mean daily protein intake, determined by individual dietary survey, at 0 and 4 weeks, remained constant in both groups. In conclusion, weight gain and IGF-1 and transferrin level increases and BUN level decreases, despite the constant oral intake, suggest that short-term rhGH administration is associated with an anabolic reaction in malnourished dialysis patients. PMID- 9740163 TI - Vitamin D receptor gene polymorphisms affect secondary hyperparathyroidism in hemodialyzed patients. AB - Adynamic bone disease unrelated to aluminum deposition, with low parathyroid hormone (PTH) levels, has increased in patients with end-stage renal failure. Some patients present with severe secondary hyperparathyroidism despite calcitriol administration and phosphate restriction. Because therapeutic and environmental factors are now similar among hemodialyzed patients, the variable incidence of secondary hyperparathyroidism may be caused by genetic heterogeneity. To examine this possibility, we analyzed restriction fragment length polymorphisms of the vitamin D receptor (VDR) gene in 877 Japanese hemodialysis patients. VDR allelic polymorphism was determined by the method of Morrison et al. Polymerase chain reaction (PCR) amplification and a BsmI endonuclease restriction site at the 5' end of the VDR gene defined BB (absence of restriction site on both alleles), Bb (heterozygous), or bb (restriction site on both alleles). The mean serum PTH level was lower in BB patients (86 +/- 102 pg/mL) than in bb patients (148 +/- 217 pg/mL; P < 0.05). The serum osteocalcin level was also lower in BB than in bb patients (P < 0.05). If results were re analyzed excluding patients with a history of dialysis exceeding 10 years or those with non-insulin-dependent diabetes mellitus (NIDDM) or who had undergone parathyroidectomy, the differences in serum PTH levels were greater. However, there was no significant difference in serum PTH levels among the VDR genotypes, only for patients with NIDDM. The present study shows that patients with the b allele for the VDR gene have more severe secondary hyperparathyroidism than patients without the b allele. However, NIDDM or a long history of hemodialysis has a stronger power to influence PTH secretion. PMID- 9740164 TI - Pain comparison after subcutaneous administration of single-dose formulation versus multidose formulation of epogen in hemodialysis patients. AB - Subcutaneous (s.c.) injection is an approved route of administration for recombinant human erythropoietin (epoetin alfa). However, pain at the injection site with the single-dose formulation has limited its use. With the recent development of a multidose formulation containing benzyl alcohol as a preservative, anecdotal reports have emerged that suggest this product causes less stinging. Using a randomized, triple-blind, placebo-controlled crossover design, this study compared pain perception, intensity, and duration after s.c. injection with a multidose formulation versus single-dose formulation using visual analogue (VAS) and verbal descriptive pain scales (VDS). Twenty-eight hemodialysis patients received s.c. injections of placebo (normal saline) in one arm and either the multidose or single-dose formulation in the opposite arm. One week later, the subjects again received placebo in one arm and the other epoetin alfa formulation in the opposite arm. The VAS and VDS measurements were obtained at time 0 and then every 5 minutes for a period of 30 minutes. Results showed a statistically significant difference in pain perception between formulations at times 0, 10, and 15 minutes for both the VAS and VDS. In conclusion, there was a significant difference in pain perception between formulations with the multidose formulation causing less pain than the single-dose formulation. However, it should be noted that several patients reported no pain with the single-dose formulation. This indicates that individual patient response could be considered when deciding which formulation to use, although it may be difficult to implement an error-free distribution and administration system using the two different formulations. PMID- 9740165 TI - Homocysteine, B vitamins, and vascular-access thrombosis in patients treated with hemodialysis. AB - To evaluate whether increased plasma homocysteine concentrations (hyperhomocysteinemia) are associated with thrombosis of arteriovenous (AV) grafts, we determined plasma homocysteine, plasma and erythrocyte folate, plasma vitamin B12, and vitamin B6 (pyridoxal-5'-phosphate [PLP]) in 48 patients (45 black patients and three white patients) with end-stage renal disease who received hemodialysis. 5,10-Methylenetetrahydrofolate reductase (MTHFR) genotypes were also analyzed. The patients were divided into two groups, including a thrombosis-prone group with frequent graft loss (n = 24) and a control group with prolonged graft survival who were matched by age and duration of dialysis (n = 24). Hyperhomocysteinemia (>15 micromol/L) was found in 42 patients. There were no significant differences in all values, including the concentrations of homocysteine and vitamins between the two groups. Based on plasma folate and PLP concentrations, over 70% of patients appeared to have inadequate folate and/or vitamin B6 nutriture. Plasma homocysteine concentrations showed significant negative correlations with plasma and erythrocyte folate, and plasma vitamin B12 in all patients combined, whereas no significant correlation was found between plasma PLP and homocysteine concentrations. Among 48 patients, the heterozygous mutation (Val/Ala) of MTHFR was found only in three patients, two of whom belonged to the thrombosis-prone group and one to the control group, and there were no individuals with homozygous thermolabile mutation (Val/Val). All three white patients had Ala/Ala genotype, and 3 in 45 black patients (6.7%) were heterozygotes (Val/Ala). PMID- 9740166 TI - Disadvantage of long-term CAPD for preserving cardiac performance: an echocardiographic study. AB - The indices of cardiac performances were compared between 31 continuous ambulatory peritoneal dialysis (CAPD) and 20 long-term hemodialysis (HD) patients. They were subdivided into three groups according to dialysis duration: L-CAPD (n = 16, mean age and CAPD duration were, respectively, 53 +/- 8 [SD] years and 77 +/- 13 months); S-CAPD (n = 15; 52 +/- 12 years, 28 +/- 12 months); HD (n = 20; 51 +/- 10 years, 162 +/- 52 months). The diabetic HD patients (DM-HD; n = 13; 60 +/- 13 years of age, 22 +/- 11 months) were chosen separately. Thirteen normotensive subjects with normal kidney function (mean age, 57 +/- 9 years) were selected as an age-matched control group. There were no significant differences between groups in age, gender, incidence of original kidney disease, or serum biochemical data. The blood pressure and the cardiothoracic ratio in L CAPD were highest among groups. The indices of left ventricular (LV) hypertrophy as well as LV performance by means of echocardiography or pulsed Doppler were compared. Among nondiabetic dialysis patients, the calculated LV mass index (LVMI) of 166.4 +/- 84.3 g/m2 and the ratio of the peak atrial filling velocity to the peak diastolic flow velocity of 1.25 +/- 0.4 in L-CAPD were greatest, and the left ventricular fractional shortening (%FS) of 34.2 +/- 10.8% in L-CAPD was smallest. LVMI or %FS of L-CAPD was the same as DM-HD of 161.0 +/- 40.7 g/m2 or 31.6 +/- 8.2%. Possibly, poor control of hypervolemia, which is caused by peritoneal problems induced by either peritonitis or chronic exposure to high glucose dialysate, causes a substantial cardiac preload leading to incipient cardiac failure in L-CAPD. According to the similar results of L-CAPD and DM-HD, it may be that hypertension, hyperlipidemia, or long-term constant glucose loading of CAPD fluids in addition to impaired glucose tolerance by chronic renal failure is more or less related to the progression of LV hypertrophy and latent cardiac dysfunction in long-term CAPD patients. In this context, CAPD of more than 5 years' duration is disadvantageous for preserving cardiac function as compared with HD. PMID- 9740167 TI - Association between human recombinant EPO and peripheral vascular disease in diabetic patients receiving peritoneal dialysis. AB - Peripheral vascular disease is a serious and frequent problem in diabetic patients. Since the beginning of the widespread use of erythropoietin (EPO), we have noted an increase in peripheral vascular disease in diabetic patients receiving peritoneal dialysis and erythropoietin. This prompted us to study the effects of erythropoietin on peripheral vascular disease in patients receiving peritoneal dialysis. We retrospectively reviewed medical records of all diabetic patients in our program who received peritoneal dialysis from 1990 to 1996. Demographic and laboratory data as well as EPO use data were collected. Hospital days and occurrence of vascular events (defined as peripheral vascular surgery, amputation, or recommendation of vascular surgery or amputation by a vascular surgeon) were determined for diabetic patients receiving peritoneal dialysis. Comparisons were made between those who received EPO and those who did not received EPO, as well as comparing vascular events in 28 patients who received peritoneal dialysis before and after beginning EPO. Patients who received erythropoietin were found to have a significantly shorter time to a first vascular event, a greater number of vascular events, and more hospital days associated with vascular disease than diabetic patients who did not receive erythropoietin. With multivariate analysis, significant risk factors for the development of peripheral vascular disease in these patients were erythropoietin use, erythropoietin dose, and smoking. Twenty-eight patients who initially performed peritoneal dialysis without receiving EPO, and later received EPO, had a significant increase in vascular events, including amputations only while receiving EPO. We found the use of erythropoietin to be associated with peripheral vascular events in diabetic patients who receive peritoneal dialysis. Further investigation is warranted. PMID- 9740168 TI - Cryofibrinogenemia: an addition to the differential diagnosis of calciphylaxis in end-stage renal disease. AB - Cryofibrinogenemia is a disorder characterized by cryoprecipitation with variable clinical presentation that was first described by Korst and Kratochvil in 1955. Cryofibrinogen is a cold insoluble complex of fibrin, fibrinogen, and fibrin split products with albumin, cold insoluble globulin, factor VIII, and plasma proteins. Cryofibrinogenemia is associated with metastatic malignancies, collagen vascular diseases, and thromboembolic disorders and may be clinically asymptomatic or present with thromboembolic phenomena of skin and viscera. The pathogenesis of cryofibrinogenemia is unknown. It may be caused by the inhibition of fibrinolysis, leading to an accumulation of cryofibrinogen. Treatment of cryofibrinogenemia may include Stanozolol, plasmapheresis, and fibrinolytics. Cryofibrinogenemia simulates calciphylaxis clinicopathologically, because both may present with skin necrosis. Calciphylaxis has been reported in end-stage renal disease, but we report the first case of cryofibrinogen in a chronic dialysis patient. We suggest that in the appropriate clinical setting, cryofibrinogenemia should be considered in the differential diagnosis of calciphylaxis, and serum cryofibrinogen levels should be measured in end-stage renal disease patients presenting with skin necrosis. PMID- 9740169 TI - Aortic valve involvement in calciphylaxis: uremic small artery disease with medial calcification and intimal hyperplasia. AB - Calciphylaxis is a rare manifestation of abnormal calcium metabolism seen in some patients with renal disease. We describe the transesophageal echocardiographic (TEE) findings in a patient with calciphylaxis. These findings included calcification of ascending aorta and aortic valve. TEE was normal before the development of calciphylaxis. PMID- 9740170 TI - Hyperreninemic hypertension secondary to a subcapsular perinephric hematoma in a patient with polyarteritis nodosa. AB - Page kidney is the name ascribed to a rare syndrome of hyperreninemic hypertension caused by unilateral compressive perinephritis. Blood or fluid that accumulates in the perinephric subcapsular space compresses the renal parenchyma leading to ischemia. This syndrome is analogous to the description of cellophane induced perinephritis by Page in 1939. Page kidney typically presents in healthy young men after blunt trauma to the flank or abdomen, although cases have been noted after medical or surgical interventions. We report a case of a Page kidney in a young man with hepatitis B virus-associated polyarteritis nodosa. The patient presented with severe hypertension, hypokalemia, hyperreninemia, and radiographic evidence of a unilateral subcapsular hematoma. Perinephric hemorrhage developed because of necrotizing vascular inflammation and spontaneous or traumatic vascular rupture. In patients who present with new-onset hypertension and hypokalemia with a history of trauma or coexisting vasculitis, the presence of a Page kidney should be considered. PMID- 9740171 TI - De novo minimal change disease. AB - Beyond the acute posttransplantation period, glomerular causes of proteinuria in the renal allograft include recurrent glomerulopathy, transplant-associated entities, and de novo disease. We present a case of de novo minimal change disease with reversible acute renal failure occurring 2.5 years posttransplantation in a 56-year-old man. The cause of end-stage renal disease in the native kidney was membranous glomerulopathy. De novo minimal change disease in the renal allograft is an extremely rare entity requiring stringent clinical pathological criteria for diagnosis. Many of the cases previously reported as de novo minimal change disease fail to meet these criteria. We review the eight reported cases that appear to fulfill a strict definition of minimal change disease in the context of the current report. PMID- 9740172 TI - Calcific uremic arteriolopathy (calciphylaxis): an evolving entity? PMID- 9740173 TI - Foscarnet crystal deposition and renal failure. PMID- 9740174 TI - Vancomycin use and antimicrobial resistance in hemodialysis centers. PMID- 9740175 TI - Microalbuminuria: accuracy or economics. PMID- 9740176 TI - The kidney in plasma cell dyscrasias: Bence-Jones cast nephropathy and light chain deposit disease. PMID- 9740177 TI - Rifampicin-associated acute renal failure. PMID- 9740178 TI - Management of intrathoracic stomach with polypropylene mesh prosthesis reinforced transabdominal hiatus hernia repair. AB - BACKGROUND: Posterior cruroplasty repair of a large paraesophageal hiatus hernia has a higher than desirable rate of recurrence attributable to the inexorable cyclic negative intrathoracic pressure of respiration and positive intraabdominal pressure produced by straining, physical exertion, and coughing. To reduce the risk of recurrence after repair of a large hiatus hernia and intrathoracic stomach, we have used posterior cruroplasty reinforced with an onlay polypropylene mesh prosthesis. This paper reviews the feasibility of this technique. STUDY DESIGN: We did a retrospective review of 44 patients with large hiatus hernia and intrathoracic stomach who had posterior cruroplasty and onlay of polypropylene mesh prosthesis applied to the crura and adjacent diaphragm to repair the hiatal defect. RESULTS: Preoperative symptoms (mean duration, 26 months) included pain (33 patients), vomiting (21), dysphagia (19) and anemia (8). The typical patient (28 men and 16 women, mean age, 60) had two-thirds or more of the stomach above the diaphragm. Organoaxial gastric volvulus and herniated large or small bowel were present in 10 and 9 patients, respectively. A gastrostomy was performed for temporary drainage in 38 patients in addition to the hernia repair; 11 patients underwent a concomitant Nissen fundoplication. Postoperative complications included pleural effusion (four patients), atrial dysrhythmia (three patients), and superficial wound infection (two patients). Mean followup for 43 patients was 52 months. There have been no clinical recurrences. CONCLUSIONS: Mesh prosthesis reinforced hiatus hernia repair is effective, appears to have a low clinical recurrence rate, and should be an option in the treatment of a large hiatus hernia with intrathoracic stomach. PMID- 9740179 TI - Laparoscopic treatment of large paraesophageal hernia with totally intrathoracic stomach. AB - BACKGROUND: Once paraesophageal hernia has been diagnosed, it should be repaired immediately because of life-threatening complications such as bleeding, ischemia, and perforation when intrathoracic strangulation or volvulus occurs. We describe our surgical strategy for treating this rare type of hiatal hernia with regard to early and late postoperative complications. STUDY DESIGN: This was a retrospective case series from a university hospital. Twelve patients (seven women and five men) with a mean age of 64 years (range, 50-76 years) and a completely intrathoracic stomach underwent laparoscopic paraesophageal hernia repair. Seven patients had a type 2 hernia, and five patients had a type 3 hernia. Additional organoaxial volvulus was present in three patients. All patients underwent reduction of the stomach and the greater omentum, excision of the hernia sac, closure of the hiatal defect, and a floppy Nissen fundoplication. RESULTS: Because of severe adhesions, one patient needed an open stomach reduction (conversion rate, 8%). The mean operating time was 161 minutes (range, 110-200 minutes), blood loss was minimal, and the mean postoperative hospital stay was 6 days (range, 4-7 days). There were no intraoperative complications, but early postoperative complications occurred in three patients (25%; one with dysphagia, 1 reoperation due to organoaxial gastric rotation with gastroduodenal obstruction, and one with deep venous thrombosis). No deaths occurred. Followup in all patients is complete, with a mean followup time of 21 months (range, 3-40 months). The complication rate after long-term followup was 8%, and reflux esophagitis symptoms in one patient were completely relieved by medical therapy. CONCLUSIONS: Laparoscopic paraesophageal hernia repair was feasible and safe with low morbidity and mortality rates in this elderly patient group. To achieve good long-term results, standard surgical treatment should include reduction of the stomach, complete excision of the hernia sac, closure of the hiatal defect, floppy Nissen fundoplication, and anterior gastropexy. PMID- 9740180 TI - Cervicothoracic approach for total mesoesophageal dissection in cancer of the thoracic esophagus. AB - BACKGROUND: The clinical significance of lymph node involvement along the recurrent laryngeal nerves in cancer of the thoracic esophagus is still controversial. Although these lymph nodes are anatomically located in a well defined compartment (proximal mesoesophagus), appropriate procedures for dissecting them are not well established. STUDY DESIGN: We retrospectively investigated clinical results over the past 10 years in 276 patients who underwent systematic dissection of cervical, mediastinal, and upper abdominal lymph nodes. We routinely performed the cervical procedure before thoracotomy for total dissection of the proximal mesoesophagus and to minimize the operative risk. RESULTS: All macroscopically recognizable lesions were resected in 94% of the patients. The hospital mortality rate was 2.5%. Recurrent nerve palsy developed in 59 patients, but it was successfully managed without prolonged hoarseness in 50 of them. The recurrent nerve node group was most frequently involved (frequency of 25% in superficial cancer, 57% in non-superficial cancer). Supradiaphragmatic lymph node involvement was limited to the recurrent nerve nodes in 25% of the patients with positive supradiaphragmatic node. The 5-year survival rate in patients with positive recurrent nerve nodes was 34%. CONCLUSIONS: Dissection of the recurrent nerve lymph nodes is essential for curative esophagectomy even in the early phase of cancer invasion. Our cervicothoracic approach for total dissection of the proximal mesoesophagus yielded acceptable outcomes. PMID- 9740181 TI - Improvement in the management of bile duct injuries? AB - BACKGROUND: Previous studies have suggested that improvements in diagnostic workup and treatment of bile duct injuries (BDI) sustained during laparoscopic cholecystectomy can be expected as experience increases with the laparoscopic procedure. Many published articles reported that early diagnosis, proper classification, and optimal timing of treatment of BDI increase the likelihood of successful treatment. This study determined whether diagnosis and management of BDI have improved over the years. STUDY DESIGN: Between June 1990 and November 1996, 106 patients were diagnosed and treated in the Amsterdam Academic Medical Center for BDI sustained during laparoscopic cholecystectomy. Detailed information was obtained about peroperative findings, time interval from laparoscopic cholecystectomy to symptoms, and interval from symptoms to diagnosis. Bile duct injuries were classified into four types. Two patient groups were compared: BDI patients diagnosed from 1990 until 1994 ("learning phase") and patients diagnosed from 1995 until 1996. RESULTS: Bile duct injuries combined with bile leakage were diagnosed significantly earlier in the second period after the learning phase. The percentages of injuries diagnosed peroperatively, "blind laparotomies," and suboptimal timed hepaticojejunostomies were not different between the groups. CONCLUSIONS: Except for earlier diagnosis of BDI in the later period than in previous years, there appeared to be no significant improvement in diagnostic workup and management during the past 2 years. PMID- 9740183 TI - Laparoscopic gastric devascularization and splenectomy for sclerotherapy resistant esophagogastric varices with hypersplenism. AB - BACKGROUND: The combination of sclerotherapy with surgical salvage for sclerotherapy-resistant esophagogastric varices has recently received much attention, however, the longterm results after such an operation have yet to be reported. This is a preliminary report of a laparoscopic adaptation of a previously described surgical procedure for the treatment of refractory esophagogastric varices. STUDY DESIGN: Laparoscopic gastric devascularization and splenectomy (Hassab's operation) was successfully performed to treat recurrent sclerotherapy-resistant giant esophageal varices (n=4) and recurrent rebleeding gastric varices (n=6). The patients included 8 men and 2 women who ranged in age from 35 to 67 years (average, 54.2 years). The procedure and clinical results were evaluated from various viewpoints. RESULTS: The duration of the operation ranged from 200 to 400 minutes (mean+/-standard deviation; 287.5+/-66.0 minutes) and blood loss from 10 to 1,500 mL (average, 515.5+/-507.9 mL). The weight of the spleen ranged from 500 to 850 g (average 608.0+/-126.6 g). Conversion to minimal open operation with a gasless lifting method was done in 1 patient because of uncontrolled bleeding from the splenic vein. There were no other major complications either intraoperatively or postoperatively. All patients had hypersplenism; preoperative platelet counts ranged from 1.6 to 6.8 x 10(4)/microL (average, 4.5+/-2.7 x 10(4) microL) and the postoperative count was from 5.9 to 36.0 x 10(4)/microL (average, 21.7+/-11.5 x 10(4) microL). Postoperative endoscopy revealed that varices disappeared, and no patient had recurrence of the varices after operation during the mean followup period of 12.8+/-4.1 months (average, 8 to 20 months). CONCLUSIONS: The combination of laparoscopic gastric devascularization and splenectomy for sclerotherapy-resistant esophagogastric varices is considered a feasible and relatively safe surgical method for patients with hypersplenism. PMID- 9740182 TI - Fulminant acute pancreatitis and infected necrosis: results of open management of the abdomen and "planned" reoperations. AB - BACKGROUND: Controversy still surrounds the management of fulminant acute necrotizing pancreatitis. Because mortality rates continue to be high, especially in patients with fulminant acute pancreatitis and infected necrosis, aggressive surgical techniques, such as open management of the abdomen and "planned" reoperations, seem to be justified. STUDY DESIGN: From 1988 through 1995, 28 patients with fulminant acute pancreatitis and infected necrosis were treated with open management of the abdomen followed by planned reoperations at our surgical intensive care unit. RESULTS: All patients had infected necrosis with severe clinical deterioration: 12 patients had an Acute Physiology and Chronic Health Evaluation (APACHE) II score > or = 20 and 16 patients had a Simplified Acute Physiology Score (SAPS) > or = 15. Nineteen patients suffered from severe multiorgan failure; the remaining 9 patients needed only ventilatory and inotropic support. The mean number of reoperations was 17. In 14 patients, major bleeding occurred; fistula developed in 7. Later, 9 abscesses were drained percutaneously. The hospital mortality rate was 39%. Longterm morbidity in survivors was substantial, especially concerning abdominal-wall defects. CONCLUSIONS: Open management of the abdomen followed by planned reoperations is an aggressive but reasonably successful surgical treatment strategy for patients with fulminant acute pancreatitis and infected necrosis. Morbidity and mortality rates were high, but in these critically ill patients, such high rates could be expected. Because management and clinical surveillance require specific expertise, management of these patients is best undertaken in specialized centers. PMID- 9740184 TI - Longterm effects of hepatic arterial interleukin-2-based immunochemotherapy after potentially curative resection of colorectal liver metastases. AB - BACKGROUND: Our previous study of hepatic arterial infusion of interleukin-2 (IL 2)-based immunochemotherapy demonstrated a high response rate of patients with unresectable liver metastases. In this study, we applied this therapy to the prevention of liver recurrence in patients who underwent potentially curative resection of liver metastases. STUDY DESIGN: A pilot study was conducted of 18 patients with liver metastases from primary colorectal cancer who underwent potentially curative liver resection followed by adjuvant immunochemotherapy. The regimen consisted of a weekly hepatic arterial infusion of IL-2 (1.4-2.1 X 10(6) U) and 5-fluorouracil (250 mg) and a bolus of mitomycin C (2-4 mg) for 6 months. RESULTS: Among 18 patients, 14 are still alive with a median postoperative survival of 52 months (as of April 1998). The 5-year overall survival rate was 75%. Although recurrent cancer developed in 6 of the 18 patients, no patients had recurrence in the residual liver. This complete prevention of liver recurrence is believed to have contributed to the high 5-year survival rate (75%) as compared with the survival rate of patients treated with surgery alone (average, 30%-40%) or with several other forms of adjuvant therapy. CONCLUSIONS: Interleukin-2-based immunochemotherapy is useful in combination with liver resection for the prevention of liver recurrence in colorectal cancer patients with liver metastases. A multicenter randomized trial is recommended. PMID- 9740186 TI - APACHE II score: a useful tool for risk assessment and an aid to decision-making in emergency operation for bleeding gastric ulcer. AB - BACKGROUND: Operating for bleeding gastric ulcer remains controversial. Gastric resection bears a higher surgical risk while limited operation may result in more postoperative hemorrhage. There has been little discussion of effective risk assessment of patients. The aim of this study is to define surgical risk by using the APACHE II scoring system, and to determine optimal management. STUDY DESIGN: Records from October 1990 to December 1996 were retrospectively reviewed for patients (n=101) with bleeding gastric ulcer who had undergone emergency operation after failed endoscopic therapy. Mortality rates were examined according to different APACHE II scores, and the surgical risk was defined. From January 1997 to December 1997, 35 consecutive patients were enrolled for prospective study. Partial gastric resection (PGR) was performed for patients with huge ulcers (>2 cm) and for low-risk patients with ulcers at the antrum or angularis, while limited operation (oversewing or excision of bleeding ulcer) was reserved for others. The results were compared with the retrospective study. RESULTS: In the retrospective study, the mortality rates for the group with a score < 15 and > or = 15 were 5% (3 of 63) and 58% (22 of 38), respectively (p < 0.05). In the group with a score < 15, PGR was performed on 27 patients, and one died. For those patients with a score > or = 15, PGR carried a lower mortality than limited operation, although this was not statistically significant (47% vs 65%). Limited operation resulted in an overall rate of 22% postoperative hemorrhage and 12% reoperation rate, in which all patients with a score > or = 15 died. In the prospective study, the mortality rates in those scoring <15 and > or = 15 were 6% and 50%, respectively. This is not significantly different than the retrospective study. However, the rate of postoperative hemorrhage was diminished (5%). CONCLUSIONS: APACHE II score is a useful tool for assessing risk in patients with bleeding gastric ulcer. The mortality is minimal in those with a score <15, and PGR can be performed with low risk. Although high-risk patients have dreadful outcomes, limited operation cannot improve them if postoperative hemorrhage occurs. Decision making in emergency operation for such patients should be based on the ulcer conditions and the patient's hemodynamic status. PMID- 9740185 TI - Attenuation of ischemic liver injury by prostaglandin E1 analogue, misoprostol, and prostaglandin I2 analogue, OP-41483. AB - BACKGROUND: Prostaglandin has been reported to have protective effects against liver injury. Use of this agent in clinical settings, however, is limited because of drug-related side effects. This study investigated whether misoprostol, prostaglandin E1 analogue, and OP-41483, prostaglandin I2 analogue, which have fewer adverse effects with a longer half-life, attenuate ischemic liver damage. STUDY DESIGN: Thirty beagle dogs underwent 2 hours of hepatic vascular exclusion using venovenous bypass. Misoprostol was administered intravenously for 30 minutes before ischemia and for 3 hours after reperfusion. OP-41483 was administered intraportally for 30 minutes before ischemia (2 microg/kg/min) and for 3 hours after reperfusion (0.5 microg/kg/min). Animals were divided into five groups: untreated control group (n=10); high-dose misoprostol (total 100 microg/kg) group (MP-H, n=5); middle-dose misoprostol (50 microg/kg) group (MP-M, n=5); low-dose misoprostol (25 microg/kg) group (MP-L, n=5); and OP-41483 group (OP, n=5). Animal survival, hepatic tissue blood flow (HTBF), liver function, and histology were analyzed. RESULTS: Two-week animal survival rates were 30% in control, 60% in MP-H, 100% in MP-M, 80% in MP-L, and 100% in OP. The treatments with prostaglandin analogues improved HTBF, and attenuated liver enzyme release, adenine nucleotrides degradation, and histologic abnormalities. In contrast to the MP-H animals that exhibited unstable cardiovascular systems, the MP-M, MP-L, and OP animals experienced only transient hypotension. CONCLUSIONS: These results indicate that misoprostol and OP-41483 prevent ischemic liver damage, although careful dose adjustment of misoprostol is required to obtain the best protection with minimal side effects. PMID- 9740187 TI - Efficient inhibition of intimal hyperplasia by adenovirus-mediated inducible nitric oxide synthase gene transfer to rats and pigs in vivo. AB - BACKGROUND: Inadequate nitric oxide (NO) availability may underlie vascular smooth muscle overgrowth that contributes to vascular occlusive diseases including atherosclerosis and restenosis. NO possesses a number of properties that should inhibit this hyperplastic healing response, such as promoting reendothelialization, preventing platelet and leukocyte adherence, and inhibiting cellular proliferation. STUDY DESIGN: We proposed that shortterm but sustained increases in NO synthesis achieved with inducible NO synthase (iNOS) gene transfer at sites of vascular injury would prevent intimal hyperplasia. We constructed an adenoviral vector, AdiNOS, carrying the human iNOS cDNA and used it to express iNOS at sites of arterial injury in vivo. RESULTS: AdiNOS-treated cultured vascular smooth muscle cells produced up to 100-fold more NO than control cells. In vivo iNOS gene transfer, using low concentrations of AdiNOS (2 x 10(6) plaque forming units [PFU]/rat) to injured rat carotid arteries, resulted in a near complete (>95%) reduction in neointima formation even when followed longterm out to 6 weeks post-injury. This protective effect was reversed by the continuous administration of an iNOS selective inhibitor L-N6-(1-iminoethyl) lysine. However, iNOS gene transfer did not lead to regression of preestablished neointimal lesions. In an animal model more relevant to human vascular healing, iNOS gene transfer (5 x 10(8) PFU/pig) to injured porcine iliac arteries in vivo was also efficacious, reducing intimal hyperplasia by 51.8%. CONCLUSIONS: These results indicate that shortterm overexpression of the iNOS gene initiated at the time of vascular injury is an effective method of locally increasing NO levels to prevent intimal hyperplasia. PMID- 9740188 TI - Leg ulceration in the sickle cell patient. AB - BACKGROUND: The purpose of this study was to determine cost of care for leg ulcers in sickle cell patients and suggest an improved modality in ulcer care. STUDY DESIGN: We performed a retrospective study of a group of sickle cell disease patients with leg ulcers. RESULTS: Eighteen patients with a leg ulcer (duration: mean, 53.7 months), sickle cell disease, and a mean of 20.7 years of age had various modalities of treatment with the only consistency in healing being a commercial moist-wound dressing. CONCLUSIONS: There is no consistency in the treatment of the sickle cell patient with a leg ulcer. Treatment with a moist dressing had the best results. PMID- 9740189 TI - The intrathoracic stomach. PMID- 9740190 TI - Treatment of primary gastric lymphoma and gastric mucosa-associated lymphoid tissue lymphoma. PMID- 9740191 TI - The role of the endothelium in changes in procoagulant activity in sepsis. PMID- 9740192 TI - End-to-end pancreaticoduodenostomy: an alternative reconstruction for partial resection of the head of the pancreas. PMID- 9740193 TI - Zuckerkandl's tuberculum: an arrow pointing to the recurrent laryngeal nerve (constant anatomical landmark) PMID- 9740194 TI - Sentinel lymph node biopsy. PMID- 9740195 TI - En bloc transplantation of infant kidneys. PMID- 9740196 TI - Detection of Helicobacter pylori infection in children with a standardized and simplified 13C-urea breath test. AB - BACKGROUND: The 13C-urea breath test, a reliable noninvasive method of detection of Helicobacter pylori in adults, needs validation in children. METHODS: In order to evaluate the diagnostic accuracy of 13C-urea breath test in children, the results of this test performed in 144 children were correlated with the histology and culture of contemporaneous gastric (antral and fundic) biopsy specimens. The test was performed with 2 mg/kg body weight 13C-Urea (maximum, 100 mg) ingested after a fat-rich test meal. Samples of expired breath taken at 0, 5, 10, 20, and 30 minutes were assayed with mass spectrometry. Results were considered positive when the curve of excretion of labeled carbon dioxide in the expired breath increased by 5%O or more above the baseline. RESULTS: Discrepancies in H. pylori status were observed in 14 children. To improve and simplify the test, the results were reanalyzed using different cutoff values for each sampling time. The best results, with sensitivity of 95.7% and specificity of 95.2%, were obtained with a cutoff of 3.5%O at 20 minutes. CONCLUSIONS: The 13C-urea breath test is a reliable method for the noninvasive detection of H. pylori infection in children. The test can be simplified and its accuracy improved using only the 0- and 20 minute breath samples and a cutoff of 3.5%O instead of the classical 5%O used in adults. The need for modification of the cutoff value may reflect the higher production of endogenous CO2 in children. PMID- 9740197 TI - Polyamines in human and rat milk influence intestinal cell growth in vitro. AB - BACKGROUND: Polyamines are required for intestinal growth and development. In this study, we examined whether milk can supply the polyamines needed for growth of IEC-6 cells, a line on non-transformed rat intestinal crypt cells. METHODS: Human, bovine, and rat milk, and cow's milk-based infant formula were studied. Human, bovine, and rat milk were defatted and sterilized by filtration. IEC-6 cells were stabilized in Dulbecco's modified Eagle's medium (DMEM) containing 0.5% fetal bovine serum, 5 mM L-glutamine, 100 U/mL penicillin and 100 microg/mL streptomycin for 24 h at 37 degrees C. Thereafter, to initiate active growth, cells were placed in fresh DMEM containing 5% FBS (plus the other ingredients) supplemented with 5% (vol/vol) milk or infant formula. In some experiments, cells were also treated with difluoromethylornithine (2.5 mM) (DFMO), an inhibitor of polyamine synthesis, or dialyzed milk plus DFMO. After 44 hours of culture, cells were pulsed with 3H-thymidine (3H-TdR) for 4 hours, harvested and the radioactivity incorporated into DNA was measured. RESULTS: Human and rat milk stimulated proliferation of IEC-6 cells (p < 0.05 compared to controls); addition of DFMO did not reverse the stimulatory effect. Bovine milk and the infant formula did not stimulate proliferation or prevent the growth inhibition induced by DFMO. After dialysis, human milk had less ability to reverse the DFMO inhibition (p < 0.05). CONCLUSIONS: These experiments suggest that both human and rat milk, but neither bovine milk nor the infant formula, contain sufficient bioactive polyamines to sustain cell growth during inhibition of polyamine synthesis. PMID- 9740198 TI - Nutrient intake and growth of infants with phenylketonuria undergoing therapy. AB - BACKGROUND: Because of reports of poor growth, a study was conducted for 6 months in 35 infants with classic phenylketonuria diagnosed during the neonatal period who were fed Phenex-1 Amino Acid Modified Medical Food With Iron (Ross Products Division, Columbus, OH, U.S.A.).as their primary protein source. METHODS: Diet diaries and anthropometric measures were obtained monthly as part of a larger study in which nutrition status was evaluated. RESULTS: In 6-month-old infants, mean percentiles for crown-heel length (59.14+/-4.31 SEM), head circumference (63.88+/-4.50) and weight (71.51+/-4.25) were normal. Mean (+/- SEM) daily intake of medical food was 79+/-4 g; protein and energy intakes were 17.3+/-0.6 g and 2772+/-75.6 kJ (660+/-18 kcal). Mean daily phenylalanine and tyrosine intakes per kilogram of body weight were 40+/-1 mg and 219+/-9 mg. Intakes of protein, energy, and tyrosine were positively correlated with crown-heel length, head circumference, and weight at 3 months of study. Overall plasma phenylalanine and tyrosine concentrations during the 6-month study were 297+/-41 micromol/l and 58+/-5 micromol/l, respectively. Neither plasma phenylalanine nor tyrosine concentration was correlated with growth. CONCLUSION: Phenex-1 supports normal growth when fed in adequate amounts. These data support those of the Medical Research Council Working Party on Phenylketonuria for 3 g/kg per day of amino acids from medical food. PMID- 9740199 TI - Helicobacter pylori-positive gastritis in pediatric patients with chronic inflammatory bowel disease. AB - BACKGROUND: Gastritis is a common finding in patients with inflammatory bowel disease. However, the association of gastritis with Helicobacter pylori is unclear in these patients. METHODS: The prevalence of antibodies for H. pylori in serum was determined in 47 pediatric patients with inflammatory bowel disease (19 with Crohn's disease, 21 with ulcerative colitis, and 7 with unclassified disease). H. pylori antibodies of the IgG and IgA classes were measured by enzyme immunoassay in 24 patients at the time of diagnosis of inflammatory bowel disease and in 23 more patients during the follow-up of inflammatory bowel disease (mean follow-up, 3.5 years; range 1-10 years). Esophagogastroduodenoscopy was performed on 40 patients during the examination for inflammatory bowel disease. RESULTS: In contrast to earlier findings, no patient was determined to be positive for H. pylori, either in serologic or histologic examination. This negative finding was unexpected, because it is known that approximately 10% of asymptomatic Finnish children have antibodies for H. pylori in serum and approximately 10% of analyses of specimens obtained in gastric antral biopsies obtained at the Hospital for Children and Adolescents, Helsinki, Finland, are positive for H. pylori. CONCLUSIONS: Permanent colonization of the stomach with H. pylori is unusual in children with inflammatory bowel disease. PMID- 9740200 TI - Faecal bile acid and dietary residue excretion in cystic fibrosis: age group variations. AB - BACKGROUND: Earlier studies report the excessive faecal excretions of bile acids and dietary residues in cystic fibrosis (CF). However, few of these investigated large groups of patients using modern pancreatin preparations and little data exists reporting carbohydrate excretion. We therefore aimed to characterise the general levels of malabsorption within age groups of 132 patients attending a regional CF centre. METHODS: The faecal excretions of bile acids, fat, nitrogen and carbohydrate were measured. Most of these patients were treated with either (Creon) (n = 58) or Pancrease (n = 51) and prophylactic antibiotics. The patients were grouped in age ranges 0.5 to 5 years, 6 to 10 years, 11 to 15 years and >16 years. Carbohydrate excretion was determined in the 11 to 15 year group. RESULTS: Increased excretions with increment in age group were found which, for bile acids, was twice that of age matched controls. Modest relationships were found between the overall excretion of bile acids and fat, and between the excretion of bile acids and nitrogen. Primary bile acids were a feature of cystic fibrosis stools but the patterns of individual bile acid excretion revealed a trend towards a normal bile acid types with increment in age group. Faecal carbohydrate was significantly increased to levels which may significantly alter large bowel microflora. CONCLUSIONS: The data adds to the evidence that maldigestion initiates bile acid sequestration and consequently, the predominance of primary bile acids. PMID- 9740201 TI - Bacterial wall lipopolysaccharide as a cause of intussusception in mice. AB - BACKGROUND: There is evidence that intussusception is associated with bacterial infection. It was hypothesized that a component of the bacterial wall may induce the intussusception. This study was intended to determine whether lipopolysaccharide from Escherichia coli or Salmonella can initiate intussusception in mice. METHODS: Lipopolysaccharide was injected intraperitoneally in mice, and the animals were examined for the presence of intussusception from 2 to 192 hours after injection. Gastrointestinal transit was assessed by measuring the passage of charcoal in the small intestine. Transit index was defined as the ratio between the distance traveled by charcoal and the total length of the small intestine. RESULTS: Intussusceptions were found in as much as 25.9% of lipopolysaccharide-injected animals, whereas in control animals, the incidence was zero. The threshold for the lipopolysaccharide effect was at 4 mg/kg and incidence reached a plateau at 8 mg/kg to 16 mg/kg. The incidence of intussusception peaked 6 hours after injection of lipopolysaccharide and declined to zero after 15 hours. To test the possibility that lipopolysaccharide induces intussusception by altering motility, its effect on transit index was measured. A dose of 12 mg/kg lipopolysaccharide reduced the transit index from 56.2+/-1.4% to 37.7+/-2.1% (p < 0.05). No microscopic histologic changes were found in the bowels with intussusception. CONCLUSIONS: Intraperitoneal bacterial wall lipopolysaccharide causes intussusception in mice by disturbing gastrointestinal motility. PMID- 9740202 TI - Population screening for neonatal liver disease: a feasibility study. AB - BACKGROUND: Extra-hepatic biliary atresia and several other causes of neonatal liver disease carry high mortality and morbidity rates, especially if not treated early in life. Despite professional recommendations, delayed referral of infants with prolonged jaundice continues to be a significant problem. One approach to reducing the age of referral and diagnosis is population screening to detect significant conjugated hyperbilirubinaemia as an index of liver dysfunction. METHODS: To investigate this possibility, and to provide reference data on bilirubin and its conjugated and unconjugated fractions in a normal newborn population, 1157 neonates were anonymously tested (median age 7 days, range 4-28 days) using surplus plasma from routinely collected neonatal screening specimens, using dry slide chemistry. RESULTS: Of 2310 specimens received, 50% were suitable for analysis. The remainder were either haemolysed or insufficient (10% and 40% of the total, respectively). Total bilirubin concentrations ranged from 9 to 428 micromol/l and conjugated bilirubin from 0 to 175 micromol/l, although the latter was rarely increased to more than 30 micromol/l (2.5th-97.5th percentile ranges 15-285 micromol/l and 0-18 micromol/l, respectively). The range of the percentage of conjugated bilirubin was 0-57% (2.5th-97.5th percentile; range 0-20%). CONCLUSION: An increased conjugated bilirubin, expressed as a concentration or as the percentage of the total bilirubin, could be used as a specific marker to screen for liver dysfunction in neonates. This approach has the potential to improve the age of referral and the prognosis of infants with neonatal liver disease. PMID- 9740203 TI - Elevated intakes of zinc in infant formulas don not interfere with iron absorption in premature infants. AB - BACKGROUND: Zinc and iron may share common pathways for absorption and compete for uptake into mucosal cells. We determined whether elevated ratios of zinc to iron would interfere with erythrocyte incorporation of iron in premature infants both during and between feeds. METHODS: In the first experiment, five premature infants (<2500 g birth weight) were enrolled, once receiving full oral feeds by nasogastric tube. They received either high (1200 microg/kg, ratio 4:1) or low (300 microg/kg, ratio 1:1) doses of oral zinc sulfate, together with 300 microg/kg oral 58Fe as chloride in saline with 10 mg/kg vitamin C, between designated feeding periods. Each infant served as its own control and randomly received either high or low doses of zinc or iron and then the alternate dose after 2 weeks. In the second experiment, nine additional premature infants were assigned to the same zinc:iron intake protocol except zinc and iron were given with usual oral feeds (premature formula or human milk) equilibrated before feeding. Iron absorption was measured by the erythrocyte incorporation of 58Fe. RESULTS: High doses of zinc given between feeds significantly inhibited erythrocyte incorporation of iron. 58Fe incorporation (%) with the 1:1 ratio of zinc:iron intake was 7.5 (5.7, 10; geometric mean, -I SD, +1 SD). The percentage of 58Fe incorporation on the 4:1 ratio of zinc:iron intake was 3.6 (2.6, 5.1). Given with feeds, the percentage of 58Fe incorporation on low zinc:iron intake was 7.0 (2.6, 19). Finally, the percentage of 58Fe incorporation on high zinc:iron intake was 6.7 (2.5, 19). CONCLUSION: Elevated intakes of zinc do not interfere with erythrocyte incorporation of iron in premature formulas. PMID- 9740204 TI - Effects of human milk pasteurization and sterilization on available fat content and fatty acid composition. AB - BACKGROUND: Human milk is frequently heat treated in hospitals to reduce bacterial contamination, particularly in banked milk fed to preterm infants. Pasteurization and sterilization may induce oxidative losses of unsaturated lipids and vitamins and may inactivate enzymes and immunologic factors. This study was designed to examine the effects of pasteurization and sterilization on milk fat content available to the recipient infant and on fatty acid composition. METHODS: In fresh, pasteurized (62.5 degrees C for 30 minutes), and sterilized (120 degrees C for 30 minutes) milk samples (5 ml) of 12 mothers (days 5-35 of lactation), fat content was determined gravimetrically and the contribution of 30 fatty acids was determined by gas-liquid chromatography. RESULTS: The coefficients of variation for measurements of milk fat content were 0.7% and of fatty acids accounting for more than 0.09% of weight, 0.1-3.0%. Available fat content was 3.1+/-1.4 g/dl (mean +/- SD) in fresh human milk and 3.1+/-1.4 g/dl (not significant) in pasteurized human milk. Fat content declined to 2.7+/-1.1 g/dl (p < 0.001 vs. fresh) in sterilized human milk, because of increased fat adherence to the container surface after sterilization. The percentage composition of saturated, monounsaturated, and polyunsaturated fatty acids of the n-6 (C18:3, C20:2, C20:3, and C22:4) and the n-3 series (C18:3 C20:5, C22:5, and C22:6) was not affected by thermal treatment. Milk sterilization caused a slight decrease of linoleic (-0.7% vs. fresh milk; p = 0,006) and arachidonic (-2,6%; p = 0.045) acids. CONCLUSIONS: Pasteurization of human milk does not influence fat content and composition, but sterilization may reduce available fat content by more than 10%, whereas there are only slight changes in fatty acid composition. PMID- 9740205 TI - Clinical quiz. Presumed Crohn's disease. PMID- 9740206 TI - Use of probiotics in childhood gastrointestinal disorders. AB - Probiotics appear to be useful in the prevention or treatment of several gastrointestinal disorders, including infectious diarrhea, antibiotic diarrhea, and traveler's diarrhea. Results of preliminary human and animal studies suggest that patients with inflammatory diseases, and even irritable bowel syndrome, may benefit from probiotic therapy. Probiotics represent an exciting therapeutic advance, although much investigation must be undertaken before their role in gastroenterology is clearly delineated. Questions related to probiotic origin, survivability, and adherence are all important considerations for further study. More important, each probiotic proposed must be studied individually and extensively to determine its efficacy and safety in each disorder for which its use may be considered. PMID- 9740207 TI - Pretransplant management and small bowel-liver transplantation in an infant with microvillus inclusion disease. PMID- 9740208 TI - Metastatic Crohn's disease of the lung. PMID- 9740209 TI - A case of solitary cavernous hemangioma of the small intestine with recurrent clinical anemic attacks in childhood. PMID- 9740210 TI - Recurrent pericarditis due to mesalamine hypersensitivity: a pediatric case report and review of the literature. PMID- 9740211 TI - Intractable infant diarrhea with epithelial dysplasia associated with polymalformation. PMID- 9740212 TI - Aganglionosis of the appendix: is it reliable for diagnosis of total colonic aganglionosis? PMID- 9740213 TI - Foregut dysmotility complicating persistent hyperinsulinaemic hypoglycaemia of infancy. PMID- 9740214 TI - Neonatal subdural transudation of total parenteral nutrition. PMID- 9740215 TI - Allergic colitis in two infants fed with an amino acid formula. PMID- 9740216 TI - Uptake of taurocholic acid in human hepatocytes isolated from livers of donors of different age. PMID- 9740217 TI - Antibody to tumor necrosis factor in the treatment of Crohn's disease. PMID- 9740218 TI - Genetic basis of Alagille syndrome deciphered. PMID- 9740219 TI - Nitric oxide and inflammatory bowel disease. PMID- 9740220 TI - Electrogastrography for evaluating neurologically impaired children with recurrent vomiting. PMID- 9740221 TI - Sorbitol in oral liquid cisapride. PMID- 9740222 TI - Overexpression of Bcl-2 in Kaposi's sarcoma-derived cells. AB - The pathogenesis of Kaposi's sarcoma (KS), a tumor of probable vascular origin, remains an enigma. It is still unclear whether KS is a true malignancy or whether it represents a reactive polyclonal process. Using both an immunohistochemical and an immunoblot approach, we found that cells derived from KS lesions express significant levels of Bcl-2, a protein known to prolong cellular viability and to antagonize apoptosis. Bcl-2 expression was found in AIDS-related KS-derived cells, as well as in cells derived from iatrogenic and sporadic KS, indicating that Bcl-2 upregulation may be important in the pathogenesis of KS regardless of its epidemiologic form. By contrast, fibroblasts and dermal microvascular endothelial, cells which are the probable vascular progenitors of KS cells, expressed low levels of Bcl-2. The expression of Bcl-2 in KS-derived cells was associated with a long-term survival in serum-deprived conditions, a situation that has been shown to induce apoptosis in various cell types. Incubation of fibroblasts or of dermal microvascular endothelial cells with KS cell-free supernatants did not enhance Bcl-2 expression, suggesting that Bcl-2 expression is not mediated by an agent released by KS cells. Analogously, KS supernatants failed to promote the viability of fibroblasts and of dermal microvascular endothelial cells cultured in serum-free conditions. Our findings suggest that the spindle cells derived from KS have a survival advantage and may adequately represent the tumor cells of KS. PMID- 9740223 TI - Induction of interleukin-6 production by ultraviolet radiation in normal human epidermal keratinocytes and in a human keratinocyte cell line is mediated by DNA damage. AB - The sunburn reaction is the most common consequence of human exposure to ultraviolet radiation (UVR), and is mediated at least in part by interleukin-6 (IL-6). The aim of this study was to determine if DNA is a major chromophore involved in the induction of IL-6 following UV irradiation of a human epidermoid carcinoma cell line (KB), and of normal human epidermal keratinocytes. We first confirmed that IL-6 release was associated with enhanced levels of IL-6 mRNA transcripts. The wavelength dependence for IL-6 release was then investigated by irradiating the cells at defined wavelengths (254, 302, 313, 334, and 365 nm) with a monochromator. The maximum effect on IL-6 release was observed at 254 nm with only low levels of induction observed at wavelengths above 313 nm. The wavelength dependence for UV-induced IL-6 release was similar to that for DNA absorption or for the induction of cyclobutane pyrimidine dimers (CPD). To determine whether UV-induced DNA damage mediated IL-6 secretion, the role of CPD was investigated by treating keratinocytes with photosomes (photolyase encapsulated in liposomes) followed by photoreactivating light. This photoreversal procedure led to a reduction in the levels of the UVC-induced secretion of IL-6, which in normal human keratinocytes was unambiguously associated with repair of CPD. We conclude that the release of IL-6 from human keratinocytes following short-wave UVC and UVB irradiation is mediated by DNA damage and that CPD play an important role in this process. PMID- 9740224 TI - MHC-dependent and -independent activation of human nickel-specific CD8+ cytotoxic T cells from allergic donors. AB - T lymphocytes are critical effectors in the pathogenesis of contact hypersensitivity. Nickel is the most common contact sensitizer in humans and nickel-specific CD4+ T helper cells have been extensively characterized. Because recent observations have suggested the activation of CD8+ T cells in murine models of contact hypersensitivity, we investigated the existence of CD8+ hapten specific T lymphocytes in patients with allergy to nickel. Nickel-specific T cell lines were generated from the peripheral blood of three allergic donors. The T cell lines were composed of a majority of CD4+ T cells, but CD8+ T cells were also present and their percentage increased with repeated in vitro stimulations. In addition to nickel-reactive helper T cell-0-type or helper T cell-2-type CD4+ T cell clones, CD8+ T cell clones could be derived from these cell lines and a total of 15 clones were further studied. Cytokine production was evaluated for 11 CD8+ T cell clones that were either cytotoxic T cell-0- or cytotoxic T cell-1 type clones. Additional effector functions were investigated on the complete panel of T cell clones. These CD8+ T cells did not only display hapten-specific proliferation, but also specific cytotoxic activities towards autologous EBV-B cells in the presence of nickel. Two different types of CD8+ T cells were characterized. Most of the clones lysed only autologous targets in the constant presence of nickel; however, one clone was able to lyse numerous targets in the presence of NiSO4, irrespective of the expression of either major histocompatibility complex class I or class II molecules. The characterization of nickel-specific cytotoxic CD8+ T cells with different requirements for nickel specific target lysis, may have important implications in the development or in the control of human contact hypersensitivity reactions to nickel in vivo. PMID- 9740225 TI - Partial purification and characterization of two distinct types of caspases from human epidermis. AB - Recent observations demonstrated that interleukin-1beta converting enzyme family proteases, now referred to as caspase family, play central roles in apoptosis, or programmed cell death. In this study, we tried to isolate and characterize epidermal caspases. By DEAE-Sephacel anion-exchange chromatography, human cornified cell extract showed two caspase-like fractions (F-I and F-II) with different substrate specificities. These were further purified by Sephacryl S 200, Mono Q ion exchange and Superose 6 gel chromatography. F-I showed a molecular weight of 30 kDa and specifically hydrolyzed acetyl-Asp-Glu-Val-Asp methylcoumarinamide, a fluorogenic substrate for caspase-3 (CPP32) with a Km value of 13.8 microM. F-I generated a characteristic 85 kDa fragment from poly(ADP-ribose) polymerase. Inhibitor susceptibility of F-I was very similar to that of caspase-3, further confirming the caspase-3-like properties of F-I. In contrast, the molecular weight of F-II was estimated to be 110 kDa, which was much higher than the other caspases. F-II equally hydrolyzed acetyl-Asp-Glu-Val Asp-methylcoumarinamide, and acetyl-Tyr-Val-Ala-Asp-methylcoumarinamide, caspase 1 (interleukin-1beta converting enzyme)-specific substrate, and was inhibited by acetyl-Tyr-Val-Ala-Asp-aldehyde and acetyl-Tyr-Val-Ala-Asp-aldehyde. Affinity labeling using biotinylated YVAD-cmk demonstrated several positive bands ranging from 25 to 35 kDa, supporting the hypothesis that F-II is a complex of multiple caspases. Reverse transcriptase-polymerase chain reaction analysis demonstrated that among known caspases tested, caspase-1, -2, -3, -4, and -7 were expressed in cultured human keratinocytes. These results suggest that multiple caspases are synthesized in human keratinocytes and are involved in terminal differentiation. PMID- 9740226 TI - Effects of ultraviolet B radiation on human Langerhans cells: functional alteration of CD86 upregulation and induction of apoptotic cell death. AB - We have recently reported that in vitro low dose of ultraviolet B radiation (UVB, 100-200 J per m2) directly impaired the antigen-presenting function of human Langerhans cells. In this study, we analyzed the effect of UVB irradiation on the Langerhans cells expression of several accessory molecules, namely CD54, CD80, and CD86. Langerhans cells phenotype was determined either immediately after UVB exposure (100 J per m2) or after a 2 d culture. No modification in cell surface antigen levels was observed immediately after irradiation. Prior UVB exposure did not modify the levels of CD80 at the Langerhans cells surface after a 2 d culture. In contrast, CD54 and, above all, CD86 expression were significantly decreased. Addition of exogenous anti-CD28 monoclonal antibodies partly restored the allostimulatory property of irradiated Langerhans cells in mixed epidermal cell-lymphocyte reaction, demonstrating that impairment of CD86 upregulation contributes to the UVB-induced immunosuppressive effect. Furthermore, we found that UVB irradiation at 200 J per m2 significantly reduced the number of viable Langerhans cells after 2 d of culture. UVB-induced cytotoxicity was due to apoptotic cell death, as demonstrated by typical morphologic alterations and by DNA fragmentation yielding a classical ladder pattern on gel electrophoresis. Interestingly, interaction of Langerhans cells with CD40-ligand transfected L cells improved the viability of irradiated Langerhans cells, counteracted the inhibition of CD86 expression, and efficiently reduced the number of apoptotic cells after a 2 d culture. Collectively, these results demonstrate that in vitro UVB exposure affects Langerhans cells via at least two distinct pathways: (i) decreased CD86 costimulatory molecule upregulation; and (ii) induction of Langerhans cells apoptosis, a phenomenon partly prevented by CD40 triggering. PMID- 9740227 TI - Differential regulation of P53 and Bcl-2 expression by ultraviolet A and B. AB - The induction of apoptosis by ultraviolet (UV) radiation and other DNA damaging agents plays a critical role in monitoring the accumulation of genetic damage and the suppression of tumor development. We hypothesize that UVA and UVB induce apoptosis by modulating balances between p53 and/or bcl-2 genes. Using MCF-7 cells that express both wild-type P53 and Bcl-2 proteins, we demonstrated that UVA and UVB induced apoptosis through regulating expression of apoptosis promoting or inhibiting genes. UVA induced immediate apoptosis and downregulated bcl-2 expression. Bcl-2 expression was reduced by approximately 40% at 4 h post 150 kJ UVA irradiation per m2 with a maximum downregulation (over 70%) at 24 h. The dose-response studies revealed that significant reduction of bcl-2 expression was observed at UVA doses ranging from 50 to 200 kJ per m2; however, p53 levels were not affected by UVA. In contrast, UVB exhibited a entirely different action than UVA in that UVB substantially induced p53 expression, but had no effect on bcl-2 expression. The induction of P53 by UVB was dose and time dependent with the maximum expression at 24 h post-2 and post-4 kJ UVB irradiation per m2. Down regulation of bcl-2 and fragmentation of DNA induced by UVA occurred earlier (approximately at 4 h) than upregulation of p53 and DNA fragmentation by UVB (12 24 h). These results suggest that UVA and UVB cause cell damage through different mechanisms and that the balances between the expression of p53 and bcl-2 may play an important role in regulating the apoptosis induced by UV irradiation. PMID- 9740228 TI - Water diffusion characteristics of human stratum corneum at different anatomical sites in vivo. AB - Despite its heterogeneity, stratum corneum (SC) has been described as a homogeneous membrane for water diffusion. We measured water flux across the SC, transepidermal water loss (TEWL), in six women, in vivo. At four anatomical sites -back, abdomen, forearm, and thigh--we took measurements during sequential tape stripping. The inverse of TEWL (1/TEWL) and removed SC thickness yielded a highly linear correlation (Pearson's r ranging between 0.88 and 0.99). Applying Fick's law of diffusion, we calculated SC thickness (H), and SC water diffusion coefficient (D). Comparing the results, SC of all women was significantly thicker (p < 0.05) at the extremities (12.7 +/- 4.2 microm, mean +/- SD, n = 12) than the abdomen (7.7 +/- 1.8 microm, n = 6). The calculated diffusion coefficient approximated 2.16 +/- 1.14x10(-9) cm2/s. Compared with the diffusion constant found for SC depleted of lipids, our value was 100-fold lower. In agreement with previous findings that intercellular lipids are a rate determining component of the SC barrier, we suggest that water diffuses mainly through the intercellular space. The calculation of H and/or D, however, is based on several variables: SC density, the water concentration difference, and the partition coefficient of water between viable epidermis and SC. The literature values vary widely. It is desirable to determine these parameters more precisely, especially if discrete differences, such as between anatomical sites, are to be revealed. PMID- 9740229 TI - Oxidative activity of the type 2 isozyme of 17beta-hydroxysteroid dehydrogenase (17beta-HSD) predominates in human sebaceous glands. AB - Sebum production is regulated by the opposing effects of androgens and estrogens. The intracrine activity of steroid metabolizing enzymes is important in regulating sebum production because these enzymes can convert weak steroids from the serum into potent androgens and estrogens within the sebaceous gland (SG). 17Beta-hydroxysteroid dehydrogenase (17beta-HSD) interconverts weak and potent sex steroids via redox reactions. In this regard, it may function as a gatekeeping enzyme regulating the hormonal milieu of the SG. Six isozymes of 17beta-HSD have been identified that differ in their substrate preference and their preference to produce weak or potent sex steroids via oxidation or reduction, respectively. The goals of this study are: (i) to identify which isozyme (s) of 17beta-HSD is active in SG; (ii) to determine if its activity differs in facial skin compared with nonacne-prone skin that may account for the regional differences in sebum production; (iii) to compare the activity of 17beta HSD in intact glands and in SG homogenates; and (iv) to determine if 13-cis retinoic acid inhibits 17beta-HSD activity. Human SG were assayed for 17beta-HSD activity using estrogens, androgens, and progestins as substrates. Oxidative activity of the type 2 isozyme predominated in all samples tested. Although transcripts for the types 1, 2, 3, and 4 isozymes were detected using reverse transcriptase-polymerase chain reaction, only mRNA for the predominant type 2 isozyme and the type 4 isozyme were detected in northern analysis. Greater reductive activity of 17beta-HSD was noted in SG from facial areas compared with nonacne-prone areas, suggesting an increased net production of potent androgens in facial areas. Oxidation was more predominant over reduction in intact SG compared with homogenized SG, thus supporting the hypothesis that 17beta-HSD protects against the effects of potent androgens in vivo. Activity of the type 2 17beta-HSD was not inhibited by 13-cis retinoic acid. In conclusion, SG possess the cellular machinery needed to transcribe the genes for the type 1-4 isozymes of 17beta-HSD. At the protein level, however, oxidative activity of the type 2 isozyme predominates, suggesting that 17beta-HSD isozyme activity may be translationally regulated. PMID- 9740230 TI - Tacrolimus ointment does not affect collagen synthesis: results of a single center randomized trial. AB - We conducted a randomized, double-blind, placebo-controlled trial to assess the atrophogenicity of tacrolimus ointment. In a combined group of atopic dermatitis patients (n = 14) and healthy volunteers (n = 12), 0.3% tacrolimus, 0.1% tacrolimus, betamethasone-valerate, and a vehicle control were applied in a randomized order to nonsymptomatic, 4 cm x 4 cm regions of abdominal skin. After 7 d of treatment under occlusion, the carboxy- and amino-terminal propeptides of procollagen I (PICP, PINP) and the amino-terminal propeptide of procollagen III (PIIINP) were measured from suction blister fluid with specific radioimmunoassays. In addition, ultrasound measurements of skin thickness were taken. Betamethasone-treated areas showed median PICP, PINP, and PIIINP concentrations of 17.0%, 17.6%, and 39.5% of the vehicle control at the end of the treatment period, respectively, whereas the 0.1% and 0.3% tacrolimus-treated areas showed median concentrations of approximately 100% of the vehicle control (p < 0.001). Betamethasone was also the only treatment to reduce skin thickness; the median decrease in skin thickness was 7.4% relative to 0.1% tacrolimus, 7.1% relative to 0.3% tacrolimus, and 8.8% relative to the vehicle control (p < 0.01). Results for atopic dermatitis patients and healthy volunteers were similar. These findings suggest that tacrolimus does not cause skin atrophy. PMID- 9740231 TI - PUVA inhibits DNA replication, but not gene transcription at nonlethal dosages. AB - The combination of psoralens and UVA radiation (PUVA photochemotherapy) is an established treatment for many skin disorders. UVA-induced psoralen-DNA interactions are assumed to contribute to the cutaneous anti-inflammatory and anti-proliferative effects of PUVA. PUVA-induced DNA modifications might interfere not only with DNA replication, but also with gene transcription of proinflammatory genes. We therefore studied the effect of PUVA on cell proliferation and on the transcription of the c-jun and intercellular adhesion molecule-1 genes in a promyelocytic (HL60) and a keratinocyte (HaCaT) cell line. PUVA inhibited cell proliferation increasingly with increasing 8-methoxypsoralen concentrations or UVA doses. The inhibition was observed at conditions not affecting cell viability up to 48 h after PUVA. In contrast, PUVA did not inhibit gene transcription at anti-proliferative, yet nonlethal conditions. Baseline and phorbol-ester induced c-jun mRNA expression was not inhibited, nor was baseline and IFN-gamma or phorbol-ester induced intercellular adhesion molecule-1 mRNA expression. In order to assess possible transcriptional effects of PUVA-generated reactive oxygen intermediates, the reactive oxygen intermediates-sensitive transcription factor nuclear factor kappaB was assayed in mobility shift experiments. Nuclear factor kappaB-specific binding activity was not induced 1-24 h after PUVA in extracts from PUVA-treated cells when compared with controls, whereas the pro-oxidant cytokine TNF-alpha caused a marked increase in nuclear factor kappaB binding. The presented data suggest that PUVA inhibits cell proliferation, but not transcription, at nonlethal PUVA conditions. Furthermore, the data do not support a major role for PUVA-generated reactive oxygen intermediates in the regulation of gene transcription. PMID- 9740232 TI - Mutations in the ferrochelatase gene of four Spanish patients with erythropoietic protoporphyria. AB - Erythropoietic protoporphyria is a hereditary disorder of porphyrin metabolism caused by mutations in the ferrochelatase gene. Ferrochelatase catalyzes the chelation of ferrous iron into protoporphyrin IX to form heme. Mutation analysis was performed in four Spanish erythropoietic protoporphyria families resulting in the identification of four different mutations in the ferrochelatase gene. Two of them were novel mutations, a missense mutation (1157 A-->C, H386P) and a frameshift mutation (843delC) found in two Spanish families, respectively. The third and the forth Spanish patients carried already published ferrochelatase gene mutations, a nonsense mutation (343C-->T, R115X) and a missense mutation (557T-->C, I186T), respectively. The newly described frameshift mutation (843delC) predicted formation of an abrupt mRNA. The deleterious effect of His386 to Pro substitution as a result of mutation 1157 A-->C on the ferrochelatase activity was investigated by expressing the mutant ferrochelatase in Escherichia coli. The mutant ferrochelatase exhibited only 0.8% of the wild-type ferrochelatase activity. Prediction of the secondary structure of ferrochelatase suggested that the H386P mutation disrupted the original alpha-helical structure by way of introducing a turn, a rather drastic structural change of the enzyme sufficient to cause activity loss. PMID- 9740233 TI - Identification and mapping of keratinocyte muscarinic acetylcholine receptor subtypes in human epidermis. AB - Acetylcholine mediates cell-to-cell communications in the skin. Human epidermal keratinocytes respond to acetylcholine via two classes of cell-surface receptors, the nicotinic and the muscarinic cholinergic receptors. High affinity muscarinic acetylcholine receptors (mAChR) have been found on keratinocyte cell surfaces at high density. These receptors mediate effects of muscarinic drugs on keratinocyte viability, proliferation, adhesion, lateral migration, and differentiation. In this study, we investigated the molecular structure of keratinocyte mAChR and their location in human epidermis. Polymerase chain reaction amplification of cDNA sequences uniquely present within the third cytoplasmic loop of each subtype demonstrated the expression of the m1, m3, m4, and m5 mAChR subtypes. To visualize these mAChR, we raised rabbit anti-sera to synthetic peptide analogs of the carboxyl terminal regions of each subtype. The antibodies selectively bound to keratinocyte mAChR subtypes in immunoblotting membranes and epidermis, both of which could be abolished by preincubating the anti-serum with the peptide used for immunization. The immunofluorescent staining patterns produced by each antibody in the epidermis suggested that the profile of keratinocyte mAChR changes during epidermal turnover. The semiquantitative analysis of fluorescence revealed that basal cells predominantly expressed m3, prickle cells had equally high levels of m4 and m5, and granular cells mostly possessed m1. Thus, the results of this study demonstrate for the first time the presence of m1, m3, m4, and m5 mAChR in epidermal keratinocytes. Because keratinocytes express a unique combination of mAChR subtypes at each stage of their development in the epidermis, each receptor may regulate a specific cell function. Hence, a single cytotransmitter, acetylcholine, and muscarinic drugs may exert different biologic effects on keratinocytes at different stages of their maturation. PMID- 9740234 TI - Apolipoprotein E is present in primary localized cutaneous amyloidosis. AB - Apolipoprotein E (apoE) is one of the amyloid associated proteins that is found in the amyloid plaque of Alzheimer's disease and systemic amyloidosis. ApoE might play an important part in the etiology of Alzheimer's disease by functioning as a "pathologic chaperone" to promote the formation of amyloid filaments. In this study, we investigated whether apoE is associated with amyloid deposits of primary localized cutaneous amyloidosis using immunohistochemistry, immunogold electron microscopy, and immunoblotting. The subjects consisted of 12 patients with lichen amyloidosus and one patient with macular amyloidosis. Light microscopically, amyloid deposits in the dermal papillae were round in shape and stained with Congo red. Immunohistochemically, apoE was detected in amyloid deposits in all the cases examined. Immunogold electron microscopy showed apoE immunoreactivity on the amyloid deposition. Immunoblots of amyloid-positive skin showed 35K and 14K proteins, which were taken to be apoE and its fragment, respectively. In normal skin extract, only the 35K protein was detected by the anti-human apoE. Moreover, the intensity of the amyloid-positive skin sample was stronger than that of the normal skin sample. Monoclonal anti-cytokeratin antibody reacted with the 45K protein of the amyloid-positive skin extract. These results indicate that apoE is a component of primary localized cutaneous amyloidosis, and that it might play an important role in primary localized cutaneous amyloidosis. PMID- 9740235 TI - Comparison of HPLC and stereologic image analysis for the quantitation of eu- and pheomelanins in nevus cells and stimulated melanoma cells. AB - The aim of the study was to compare two methods of quantitating eumelanins and pheomelanins, pigments synthesized by melanocytes. One is based on the high performance liquid chromatography quantitation of specific degradation products of each melanin type. The other requires image analysis, transmission electron microscopy, and stereology. In a previous study, we showed good correlations between both methods for total melanin but not for eumelanins or pheomelanins. We describe here the same comparison in more pigmented cells (nevus cells and stimulated HBL melanoma cells). Transmission electron microscopy micrographs were image analyzed to generate several primary parameters. Stereology was used for estimating melanosomal maturation, intracellular melanin content, and the number of melanized melanosomes per cell, for total melanin, eumelanins, or pheomelanins. Our results showed a good correlation between both methods for total melanin, eumelanins, and pheomelanins with an r equal to 0.99, 0.91, and 0.93, respectively, when all the points were used in the linear regression analyses. In the melanoma cell group (HBL cells cultured in media of different compositions), the chemical and morphometric estimations were not parallel in the case of eumelanins and pheomelanins. In addition, the stereologic and high performance liquid chromatography pheomelanins to eumelanins ratios were still not correlated. These results demonstrate the relevancy of the stereologic method, but the low level of melanization, the possible lack of specificity of melanogenesis in melanoma cells, and a problem of sensitivity of the stereologic method in this context seem to be obstacles in obtaining better results. The utilization of normal human melanocytes could give some answers to our hypotheses. PMID- 9740236 TI - Ligands and activators of nuclear hormone receptors regulate epidermal differentiation during fetal rat skin development. AB - Because a protective barrier is essential for life, the development of the epidermis and stratum corneum must be completed prior to birth. The epidermal permeability barrier is comprised of corneocytes embedded in a lipid enriched matrix. Recent studies from our laboratory, using an explant model of fetal rat skin development that closely parallels in utero development, have shown that hormones and other activators of members of the nuclear receptor family regulate permeability barrier ontogenesis by stimulating lipid metabolism and the formation of the extracellular lipid lamellae. Using this model we sought to determine whether these hormones and nuclear activators also regulate keratinocyte differentiation during fetal development. Profilaggrin/filaggrin and loricrin expression, assessed by in situ hybridization and by immunohistochemistry, were progressively increased during epidermal ontogenesis. Whereas profilaggrin/filaggrin and loricrin were not expressed at day 17 of gestation, by day 19 both were present in the upper layers of the epidermis and both became still more abundant by day 21. These developmental changes also occurred in fetal skin explants cultured in vitro for 4 d, although the expression levels did not appear as robust as in utero. Whereas neither profilaggrin/filaggrin nor loricrin were expressed in control explants cultured for 2 d, they were seen in explants treated with either thyroid hormone, glucocorticoids, or estrogens. In contrast, dihydrotestosterone treatment delayed the expression of profilaggrin/filaggrin and loricrin. Moreover, both clofibrate, a peroxisome proliferator-activated receptor-alpha ligand, and juvenile hormone III, a farnesoid X-activated receptor activator, markedly accelerated fetal epidermal differentiation, stimulating both profilaggrin/filaggrin and loricrin expression. Our results demonstrate that several hormones and activators of nuclear hormone receptors regulate epidermal differentiation during fetal development, affecting key constituents of both keratohyalin granules and the cornified envelope. Thus, a variety of ligands/activators of nuclear receptors accelerate not only permeability barrier ontogenesis, but also the expression of structural proteins essential for stratum corneum formation. PMID- 9740237 TI - Retinoic acid receptors regulate expression of retinoic acid 4-hydroxylase that specifically inactivates all-trans retinoic acid in human keratinocyte HaCaT cells. AB - Tissue levels of all-trans retinoic acid (RA) are maintained through coordinated regulation of biosynthesis and breakdown. The major pathway for all-trans RA inactivation is initiated by 4-hydroxylation. A novel cytochrome P-450 (CYP26) that catalyzes 4-hydroxylation of all-trans RA has recently been cloned. We have investigated regulation and properties of RA 4-hydroxylase in immortalized human keratinocyte HaCaT cells. In the absence of added retinoid, RA 4-hydroxylase (CYP26) mRNA and protein were minimally detected. Addition of all-trans RA rapidly induced RA 4-hydroxylase mRNA (within 2 h) and activity (within 6 h). Induction of both mRNA and activity was transient, returning to baseline within 48 h, and completely dependent on mRNA synthesis (i.e., blocked by actinomycin D). The synthetic retinoid CD367, which specifically activates nuclear RA receptors, also rapidly induced RA 4-hydroxylase activity. This induction, however, unlike that of all-trans RA, was long-lived (>48 h). This difference was attributable to lack of metabolic inactivation of CD367 in HaCaT cells. CD2665, which inhibits RA receptor-dependent gene transcription, blocked retinoid induction of RA 4-hydroxylase, indicating that it is mediated by RA receptors. Addition of excess unlabeled substrates specific for 10 distinct mammalian P-450 subfamilies did not compete with all-trans RA for RA 4-hydroxylase activity. RA 4 hydroxylase did not hydroxylate 9-cis RA or 13-cis RA. Inhibition of RA 4 hydroxylase activity by ketoconazole potentiated activation of RA receptors by all-trans RA. In summary, RA 4-hydroxylase is a unique, highly specific cytochrome P-450 isoenzyme, whose expression is regulated by its natural substrate, all-trans RA, through activation of RA receptors. RA 4-hydroxylase functions to limit the levels, and thereby the biologic activity of all-trans RA in HaCaT cells. PMID- 9740238 TI - Glucocorticoid deficiency delays stratum corneum maturation in the fetal mouse. AB - The stratum corneum (SC) matures during late gestation in man and other mammals. Using the fetal rat as an experimental model, we have previously shown that glucocorticoids given in pharmacologic doses accelerate fetal SC maturation and barrier formation. To determine whether glucocorticoids are required for normal SC maturation, we examined the epidermal morphology of glucocorticoid-deficient (C-) murine pups, derived from matings of mice homozygous for null mutations of the corticotropin-releasing hormone alleles. In control pups on day 17.5 of gestation (term is 19.5 d), a multilayered SC was present and neutral lipid deposition in a membrane pattern was observed using Nile red fluorescence histochemistry. Ultrastructurally, mature lamellar unit structures predominate in the SC intercellular domains. In contrast, in C-pups only a single layer of SC was evident on day 17.5, and secreted lamellar material was not organized into mature lamellar structures. Furthermore, the expression of structural proteins necessary for cornified envelope formation, involucrin, loricrin, and filaggrin, and the activity of the lipid synthetic enzymes beta-glucocerebrosidase and steroid sulfatase, markers of barrier maturation, were reduced in day 17.5 C pups. C-pups derived from pregnancies supplemented with physiologic amounts of cortisone, however, display normal SC ultrastructure on day 17.5 of gestation. Furthermore, at birth, both control and C-pups exhibit a multilayered SC replete with mature lamellar membrane structures. These data demonstrate that fetal glucocorticoid deficiency delays SC maturation, and suggests that normal levels of glucocorticoids are not absolutely required for SC development. PMID- 9740239 TI - Photocarcinogenesis and susceptibility to UV radiation in the v-Ha-ras transgenic Tg.AC mouse. AB - The v-Ha-ras transgenic Tg.AC mouse line has proven to be a useful model for the study of chemical carcinogenic potential. We undertook experiments designed to study the effect of the physical carcinogen, UV radiation, on tumorigenesis in this mouse strain. Following a total of three exposures on alternating days to a radiation source covering a cumulative UVR exposure range of 2.6-42.6 kJ per m2, squamous papillomas developed by 4 wk after initial exposure in a dose-dependent manner. Malignancies developed within 18-30 wk following the initial UVR exposure and were all diagnosed as squamous cell carcinoma or spindle cell tumors. In contrast to other mouse stains used in photocarcinogenesis studies, few p53 mutations were found in Tg.AC malignancies upon polymerase chain reaction-single stranded conformational polymorphism analysis of exons 4-8 followed by sequencing of suspicious bands; however, all tumors analyzed by in situ hybridization expressed the v-Ha-ras transgene. Immunohistochemical analysis of UVR-exposed skin taken 24 h after the last of three exposures (13.1 kJ per m2 total UVR) showed expression of p53 in hair follicles and in interfollicular epidermis, which indicates that the gene was functional. Thus, although there are some differences between the Tg.AC and other mouse models, these results suggest that the Tg.AC mouse may be a useful model for the study of acute exposure photocarcinogenesis. PMID- 9740240 TI - Role of Staphylococcus aureus surface-associated proteins in the attachment to cultured HaCaT keratinocytes in a new adhesion assay. AB - Colonization of human skin with Staphylococcus aureus is a common feature in a variety of dermatologic diseases. In order to reproducibly investigate the adherence of Staphylococcus aureus to human epidermal cells, an in vitro assay was established using the biotin/streptavidine labeling system and the HaCaT cell line. This assay was used to define the role of several Staphylococcus aureus surface proteins with regard to their function in the staphylococcal adhesion process. Our studies included the standard laboratory strain Newman as well as its genetically constructed mutants DU5873, DU5852, DU5854, and DU5886 generated by allele replacement or transposon mutagenesis, which are deficient in the elaboration of staphylococcal protein A (spa), clumping factor (clfA), coagulase (coa), and the fibronectin-binding proteins A and B (fnbA/B), respectively. In comparison with strain Newman all mutants showed remarkably reduced adherence to the HaCaT keratinocyte cell line in our assay, yielding only between 43% and 60% of the adherence capacity of strain Newman after 60 min. Bacterial adherence could be re-established by introducing the cloned wild-type genes for the surface proteins on shuttle plasmids into the chromosomally defective mutants, thus suggesting a pathogenetic role of these proteins in the attachment of Staphylococcus aureus to human keratinocytes. Bacterial adherence was additionally enhanced by alkaline pH-values that are characteristic for skin conditions with epidermal barrier dysfunction. The use of Staphylococcus aureus mutant strains, deficient in the elaboration of defined proteins, allows specific investigation of colonization and virulence factors of this dermatologic relevant microorganism. PMID- 9740241 TI - A pulsed electric field enhances cutaneous delivery of methylene blue in excised full-thickness porcine skin. AB - We used electric pulses to permeabilize porcine stratum corneum and demonstrate enhanced epidermal transport of methylene blue, a water-soluble cationic dye. Electrodes were placed on the outer surface of excised full-thickness porcine skin, and methylene blue was applied to the skin beneath the positive electrode; 1 ms pulses of up to 240 V were delivered at frequencies of 20-100 Hz for up to 30 min. The amount of dye in a skin sample was determined from absorbance spectra of dissolved punch biopsy sections. Penetration depth and concentration of the dye were measured with light and fluorescence microscopy of cryosections. At an electric exposure dose VT (applied voltage x frequency x pulse width x treatment duration) of about 4700 Vs, there is a threshold for efficient drug delivery. Increasing the applied voltage or field application time resulted in increased dye penetration. Transport induced by electric pulses was more than an order of magnitude greater than that seen following iontophoresis. We believe that the enhanced cutaneous delivery of methylene blue is due to a combination of de novo permeabilization of the stratum corneum by electric pulses, passive diffusion through the permeabilization sites, and electrophoretic and electroosmotic transport by the electric pulses. Pulsed electric fields may have important applications for drug delivery in a variety of fields where topical drug delivery is a goal. PMID- 9740242 TI - Predominant expression of CD44 splice variant v10 in malignant and reactive human skin lymphocytes. AB - The remarkable functional diversity of the cell surface receptor CD44 may be due to expression of multiple variant isoforms generated by alternative splicing of variant exons. Functional and correlative data implicate a role of CD44 variant isoforms in adhesion dependent processes such as lymphocyte recirculation and tumor progression and metastasis. We have analyzed 25 primary cutaneous lymphomas and 35 reactive lymphoid cell skin infiltrates or T cell-mediated skin diseases for the expression of CD44 variant isoforms. Irrespective of histologic typing, staging, and grading, cutaneous lymphomas as well as nonmalignant skin infiltrating CD3+ CD4+ and CD8+ T and CD19+ B lymphocytes exhibited a strong expression of CD44v10 and a moderate expression of CD44v3 as determined by immunohistochemistry, immunofluorescence microscopy, and mRNA analysis. Expression of v5, v6, v7, and v9-containing CD44 variant isoforms was not detected. Furthermore, flow cytometry revealed expression of CD44v10 on a significant proportion of peripheral blood lymphocytes from Sezary's syndrome patients and a remarkable co-expression with cutaneous lymphocyte antigen. These results indicate a distinct CD44 variant isoform expression pattern associated with skin-homing lymphocytes different to lymphatic cells at noncutaneous sites. This differential expression pattern of CD44 variant isoforms may contribute to the development of lymphocyte skin infiltrates and/or the unique biologic behavior of cutaneous lymphomas. PMID- 9740243 TI - Thymidine dinucleotides induce S phase cell cycle arrest in addition to increased melanogenesis in human melanocytes. AB - Although the induction of pigmentation following exposure of melanocytes to ultraviolet light in vivo and in vitro is well documented, the intracellular mechanisms involved in this response are not yet fully understood. Exposure to UV B radiation leads to the production of DNA damage, mainly cyclobutane pyrimidine dimers, and it was recently suggested that the thymidine dinucleotide pTpT, mimicking small DNA fragments released in the course of excision repair mechanisms, could trigger melanin synthesis. We now report that the thymidine dinucleotide pTpT induces melanogenesis both in human normal adult melanocytes and in human melanoma cells. Thus, the SOS-like response suggested by Gilchrest's work to be evolutionary conserved, based primarily on work in murine cells and guinea pigs, is also apparently present in the human. Thymidine dinucleotide is nontoxic to melanoma cells and does not induce apoptosis in these cells, but induces S phase cell cycle arrest and a proliferation slow down. Because thymidine excess in culture medium leads to the synchronization of cells in S phase, we investigated whether this phenomenon was involved in the increase in melanin synthesis. We show that melanin synthesis is specifically triggered by the dimeric form of the thymidine and not by the monomeric form pT. Thus, our data strongly support that thymidine dinucleotides pTpT mimic at least part of the effects of ultraviolet irradiation, and may hence represent an invaluable model in the study of the molecular events involved in melanogenesis induction triggered through DNA damage. PMID- 9740244 TI - Dermal fibroblasts actively metabolize retinoic acid but not retinol. AB - The accessibility of plasma retinol and retinoic acid to the epidermis may be influenced by the number and metabolic capacity of fibroblasts in papillary dermis. The metabolism of retinol-binding protein-bound all-trans-retinol, and albumin-bound all-trans-retinoic acid, by fibroblasts cultured on plastic dishes or in type I collagen gels, was examined. There were no significant differences in the metabolism of either retinoid by fibroblasts as a function of culture condition. There were large differences between retinoids, however. Retinoic acid was rapidly taken up and metabolized to unidentified polar metabolites that were released to the medium. Metabolic capacity was not saturated up to a medium retinoic acid concentration of 1 microM, and was induced further by prior exposure to retinoic acid. In contrast, retinol, although readily taken up, was not metabolized, i.e., neither retinoic acid nor retinyl ester was formed. By immunohistochemistry, the average number of fibroblasts in a 100 microm thickness of papillary dermis was estimated to be 1 x 10(6) cells per cm2. Utilizing this value, the capacity of dermal fibroblasts to metabolize retinoic acid based on fibroblast abundance in the dermis was calculated. The results suggest that fibroblasts could limit delivery of plasma retinoic acid but not retinol to the epidermis on the basis of their metabolic capacity and abundance in the dermis. PMID- 9740246 TI - Retrovirus-mediated transduction of porcine keratinocytes in organ culture. AB - Direct transfer of new genetic information to keratinocytes in epidermis may prove effective in treating certain genodermatoses; however, current methods for in vivo gene transfer to skin do not lead to persistence of the transgene. The goal of this study was to explore direct gene transfer using retrovirus-mediated transduction. Retroviral vectors integrate a DNA copy of their genome into the host chromosome and therefore have the potential to effect a permanent gene therapy. To facilitate development of methods for in vivo transduction with retroviral vectors, a porcine skin organ culture model was constructed in which the denuded surface was repopulated with replicating keratinocytes from hair follicles and epidermal remnants. In situ transduction was carried out by topical application of two retrovirus vectors, MFGlacZ (10(7) blue forming units per ml) and LZRN pseudotyped with the G protein of vesicular stomatitis virus (VSV) (10(9) colony forming units per ml), each encoding the beta-galactosidase reporter gene and the latter encoding the neomycin phosphotransferase selectable gene. Beta-galactosidase expressing cells were observed more frequently with LZRN than with MFGlacZ; however, transduction efficiency remained low in both instances. At equivalent titers, the VSV-G pseudotyped retroviral vector was shown to transduce porcine keratinocytes more efficiently than a similar vector with the amphotropic envelope. The number of beta-gal+ cells in organ culture could be increased by selection of LZRN-transduced cells in situ with G418. To achieve transduction of epidermis in vivo, these studies point out the importance of high titer retroviral vectors, pseudotyping with VSV-G protein, and in situ selection. PMID- 9740245 TI - Identification and sequencing of a putative variant of proopiomelanocortin in human epidermis and epidermal cells in culture. AB - Proopiomelanocortin (POMC) is a precursor polypeptide for various bioactive peptides, including adrenocorticotropic hormone, alpha-, beta-, and gamma melanotropin, beta-endorphin, and beta-lipotropin. Although the classical source of POMC is the pituitary, various studies indicate the expression of POMC in several nonpituitary tissues. In this study, in situ hybridization with anti sense cRNA riboprobe was used to show expression of POMC mRNA in human epidermis and cultured human epidermal cells (melanocytes and keratinocytes). POMC mRNA was amplified by reverse transcriptase-polymerase chain reaction using anti-sense and sense primers designed from Exons 2 and 3 of POMC gene. A approximately 300 bp product was present in normal human skin, grafted human skin, and cultured normal human melanocytes and keratinocytes. By Southern analysis this product was hybridized specifically to the POMC cDNA. Sequence analysis of the reverse transcriptase polymerase chain reaction product from tissues or cells showed 85% homology to POMC cDNA from human, bovine, pig, and monkey sources. This suggests the existence of a putative isoform or variant of POMC mRNA in human epidermis. PMID- 9740247 TI - Higher frequency of selective losses of HLA-A and -B allospecificities in metastasis than in primary melanoma lesions. AB - Expression of HLA class I molecules is essential for the recognition of tumor cells by CD8+ T cells. In this study, 48 bioptic samples of 42 patients in all stages of melanoma were investigated after short-time cultivation of tumor cells. To confirm melanocytic origin of cultured cells, samples were screened for mRNA expression of melanoma markers gp100, tyrosinase, MAGE-3, MelanA, and MUC18 by reverse transcriptase-polymerase chain reaction. Surface expression of specific HLA-A and -B allospecificities on melanoma cells were analyzed with a standard microcytotoxicity assay after stimulation with interferon (IFN)-alpha and compared with the background found in peripheral blood mononuclear cells from the corresponding patients. Immunohistochemistry and flow cytometry confirmed specific losses in cases where the appropriate monoclonal antibodies were available. The level of expression of HLA-I, HLA-II, and intercellular adhesion molecule 1 antigens on melanoma cells cultured in the presence or absence of IFN alpha and IFN-gamma was determined cytofluorometrically. All cell cultures tested were found to be positive for one or more melanocytic markers by reverse transcriptase-polymerase chain reaction. The specific HLA-I alleles on the cultured cells were detectable in 45 of 48 samples. In 11 cases a specific loss of one HLA-I allele was observed (2 x A2, B7, B8, B18, 4XB44, B47, B49). Ten of these samples were derived from locoregional lymphnode metastases or from distant metastatic tumors. Only one sample from a primary melanoma showed a specific loss of HLA-I (B47). IFN-alpha upregulated expression of HLA-I up to 4-fold. IFN-gamma enhanced the appearance of HLA-II up to 35-fold and the expression of intercellular adhesion molecule 1 up to 40-fold. Selective loss of HLA-I allospecificities might be a major step in tumor progression. PMID- 9740248 TI - Erythema multiforme associated human autoantibodies against desmoplakin I and II: biochemical characterization and passive transfer studies into newborn mice. AB - The demonstration of circulating autoantibodies directed against the constitutive desmosomal plaque proteins desmoplakin (dp) I and II in mucocutaneous lesions in a subset of patients with erythema multiforme major, suggests that humoral immune mechanisms may play a role in the pathogenesis of this severe skin disease. In this study we identified a specific peptide sequence--YSYSYS--representing an antigenic binding site for the human autoantibodies. This epitope is localized at the extreme carboxy terminal domain of dp thought to be responsible for the assembly of keratin filaments with desmosomes. To test the possibility whether these antibodies may exert any pathologic effects in vivo, human autoantibodies were affinity purified on a corresponding synthetic peptide matrix and peptide specific antibodies were raised in rabbits. After repeated subcutaneous injections into newborn mice, affinity-purified human autoantibodies and anti peptide rabbit IgG were detected on desmosomal plaques of keratinocytes overlying the injection site. Histologic and electron microscopic examinations showed hydropic degeneration of basal and suprabasal keratinocytes, dyskeratosis, signs of suprabasal acantholysis, and keratin filaments detached from the desmosomal plaques clumping around the nucleus. We demonstrate that autoantibodies are directed to an epitope within a dp domain crucial for the interaction of keratin filaments with desmosomes, and, when injected subcutaneously into newborn mice, produce pathologic changes. These findings imply that autoantibodies to dp could impair the function of desmosome-keratin filament complexes suggesting a pathogenic role in vivo. PMID- 9740249 TI - Perturbed epidermal pterin metabolism in Hermansky-Pudlak syndrome. AB - In Hermansky-Pudlak Syndrome (HPS) a mutation in a 79.3 kDa transmembrane protein has been shown. The function of this protein has escaped definition so far. This study unveils a defective (6R)-L-erythro-5,6,7,8-tetrahydrobiopterin (6BH4) de novo synthesis/recycling for this cofactor in HPS, where activities of the key enzyme GTP-cyclohydrolase I are in the normal range, but total biopterin levels are significantly decreased in homozygotes (n = 5) compared with unaffected controls (n = 4) (p = 0.00001). Phenylalanine hydroxylase and 4a-hydroxy-6BH4 dehydratase activities are significantly lower. mRNA of all enzymes involved in 6BH4 biosynthesis/recycling and GTP-cyclohydrolase I feedback regulatory protein were expressed in keratinocytes from homozygotes, heterozygotes, and healthy controls. Thioredoxin/thioredoxin reductase can directly control the redox status of 6BH4. These activities are allosterically controlled by calcium. Therefore calcium would directly affect this redox status. In HPS these enzyme activities are low concomitant with a defective calcium uptake, suggesting an extracellular accumulation of this second messenger. In this context phenylalanine hydroxylase is subject to phosphorylation/activation by calcium/calmodulin activated kinases. Therefore it was anticipated that calcium could directly affect the cellular L phenylalanine turnover to L-tyrosine. A significantly more rapid L-phenylalanine uptake and its turnover to L-tyrosine was identified in normal human melanocytes (n = 5) and keratinocytes (n = 2), and was more enhanced in melanocytes in the presence of 2 x 10(-3) M calcium. The turnover to L-tyrosine was significantly slower. Based on all evidence to date, we speculate that the mutated protein in HPS could be primarily involved in maintaining calcium homeostasis in this patient group. PMID- 9740250 TI - Expression of epidermal keratins and the cornified envelope protein involucrin is influenced by permeability barrier disruption. AB - In previous studies we have shown that experimental permeability barrier disruption leads to an increase in epidermal lipid and DNA synthesis. Here we investigate whether barrier disruption also influences keratins and cornified envelope proteins as major structural keratinocyte proteins. Cutaneous barrier disruption was achieved in hairless mouse skin by treatments with acetone +/- occlusion, sodium dodecyl sulfate, or tape-stripping. As a chronic model for barrier disruption, we used essential fatty acid deficient mice. Epidermal keratins were determined by one- and two-dimensional gel electrophoresis, immunoblots, and anti-keratin antibodies in biopsy samples. In addition, the expression of the cornified envelope proteins loricrin and involucrin after barrier disruption was determined by specific antibodies in human skin. Acute as well as chronic barrier disruption resulted in the induction of the expression of keratins K6, K16, and K17. Occlusion after acute disruption led to a slight reduction of keratin K6 and K16 expression. Expression of basal keratins K5 and K14 was reduced after both methods of barrier disruption. Suprabasal keratin K10 expression was increased after acute barrier disruption and K1 as well as K10 expression was increased after chronic barrier disruption. Loricrin expression in mouse and in human skin was unchanged after barrier disruption. In contrast, involucrin expression, which was restricted to the granular and upper spinous layers in normal human skin, showed an extension to the lower spinous layers 24 h after acetone treatment. In summary, our results document that acute or chronic barrier disruption leads to expression of keratins K6, K16, and K17 and to a premature expression of involucrin. We suggest that the coordinated regulation of lipid, DNA, keratin, and involucrin synthesis is critical for epidermal permeability barrier function. PMID- 9740251 TI - A novel mutation in the L12 domain of keratin 5 in the Kobner variant of epidermolysis bullosa simplex. AB - We have identified a novel mutation within the linker L12 region of keratin 5 (K5) in a family with the Kobner variant of epidermolysis bullosa simplex. The pattern of inheritance of the disorder in this family is consistent with an autosomal dominant mode of transmission. Affected individuals develop extensive and generalized blistering at birth or early infancy but in later years clinical manifestations are largely confined to palmoplantar surfaces. Direct sequencing of polymerase chain reaction products revealed a T to C transition within codon 323 of K5 in affected individuals, resulting in a valine to alanine substitution of the seventh residue within the L12 linker domain. This mutation was not observed in unaffected family members or in 100 K5 alleles of unrelated individuals with normal skin. The other critical regions of K5 and K14 were unremarkable in this family except for common polymorphisms that have been previously described. The valine at position 7 of the L12 domain is absolutely conserved in all type II keratins, and in other intermediate filament subunits as well, which suggests that this residue makes an important contribution to filament integrity. Secondary structure analysis revealed that alanine at this position markedly reduces both the hydrophobicity and the beta-sheet nature of the L12 domain. This is the first report of a mutation at this position in an intermediate filament subunit and reinforces the importance of this region to filament biology. PMID- 9740252 TI - Novel homozygous and compound heterozygous COL17A1 mutations associated with junctional epidermolysis bullosa. AB - Junctional epidermolysis bullosa is a heritable, heterogeneous blistering skin disease with mechanically induced dermal-epidermal separation, mild skin atrophy, nail dystrophy, and alopecia. Four unrelated junctional epidermolysis bullosa families with different phenotypes were investigated here and four novel mutations associated with the disease were identified. Patients 1, 2, and 3 had generalized atrophic benign epidermolysis bullosa, with nonscarring blistering and varying degree of alopecia. Patient 4 had the localisata variant of junctional epidermolysis bullosa, with predominantly acral blistering and normal hair. All patients had mutations in the COL17A1 gene encoding collagen XVII, a hemidesmosomal transmembrane protein. Patients 1 and 2 carried homozygous deletions 520delAG and 2965delG, respectively. Patient 3 was compound heterozygous for a missense and a deletion mutation (G539E and 2666delTT), and patient 4 was heterozygous for a known mutation R1226X. The deletions led to premature termination codons and to drastically reduced collagen XVII mRNA and protein levels, consistent with the absence of the collagen in generalized atrophic benign epidermolysis bullosa skin. The missense mutation G539E allowed synthesis of immunoreactive collagen XVII in keratinocytes, but prevented its secretion, thus causing lack of the protein in the skin. The data suggest that different COL17A1 mutations and their combinations can result in a spectrum of biologic and clinical phenotypes of not only generalized atrophic benign epidermolysis bullosa, but also localized junctional epidermolysis bullosa. PMID- 9740254 TI - Non-invasive (real-time) imaging of histologic margin of a proliferative skin lesion in vivo. PMID- 9740253 TI - Novel COL7A1 mutations in dystrophic forms of epidermolysis bullosa. AB - Mutations in the type VII collagen gene (COL7A1) have been shown to underlie different variants of dystrophic epidermolysis bullosa (DEB). Examination of the genetic database indicates that most of the mutations are family specific, with few recurrent mutations. To facilitate further refinement of genotype/phenotype correlations in DEB, we have examined a cohort of nine families with DEB (seven recessively and two dominantly inherited) by a mutation detection strategy based on polymerase chain reaction amplification of COL7A1 genomic sequences, followed by heteroduplex scanning and direct nucleotide sequencing. The results revealed 16 allelic mutations, 11 of them being novel, previously unpublished. The genetic information was also used for prenatal testing in a family at risk for recurrence of a severe, Hallopeau-Siemens type of RDEB. These data contribute to the expanding database of COL7A1 mutations in DEB. PMID- 9740255 TI - Somatic mutations of the MEN1 tumor suppressor gene detected in sporadic angiofibromas. PMID- 9740256 TI - Erythropoietic protoporphyria: a new mutation responsible for exon skipping in the human ferrochelatase gene. PMID- 9740257 TI - Is human papillomavirus type 5 the putative autoantigen involved in psoriasis? PMID- 9740258 TI - SEAMEO TROPMED after 30 years. PMID- 9740259 TI - Entomological and epidemiological investigations of malaria transmission in relation to population movements in forest areas of north-west Thailand. AB - Transmission of forest-related malaria was observed entomologically and epidemiologically for 2 transmission seasons in 1990 and 1991 in 5 villages of Mae Sariang district, Mae Hong Son Province, north-west Thailand. The entomological study included collections of mosquitos and determination of infection rate by using enzyme-linked immunosorbent assay in the residential villages and the farm huts. The epidemiological study included fortnightly visits to 30% of the households to interview and record movement activities and illness of villagers. Circumsporozoite proteins, in most cases of Plasmodium falciparum, were detected in Anopheles minimus species A, An. dirus s.l., An. maculatus s.s. and An. sawadwongporni in residential villages and/or farm huts, suggesting transmission could occur there. Movement of people away from their residences occurred throughout the year for several reasons with a sharp peak in July for agricultural activity, mainly ploughing and planting for rice cultivation. The relative risk of infection for people engaged in agricultural activity was 3 times that of people living in the residential villages. Although a higher biting density of vectors was generally evident at the farm huts, the estimated inoculation rates in the 2 settings were similar. Movement for forest activity increased after harvesting rice in the cool dry season and carried the highest malaria risk, suggesting different epidemiological and probably entomological conditions which need further investigation. The significance is discussed of discrepancies between the case classification system used by this study and that used by malaria sector staff. PMID- 9740260 TI - Studies on malaria during pregnancy in a tribal area of central India (Madhya Pradesh). AB - In tribal villages of central India where malaria is highly prevalent (mesoendemic), this preliminary study was undertaken to determine the effects of malaria infection in a group of 456 pregnant women with or without fever. Only 96 women were found infected with malaria, of which Plasmodium falciparum accounted for 64% of the detected parasites, while P. vivax for the remaining 36%. There were no instances of cerebral malaria or death however, one abortion and four still births were recorded among 38 primigravid women. Only one neonate was found infected with P. falciparum on day 21 though parasitemia was not high. Anemia was commonly present in most of the women (80%). Failure to clear P. falciparum parasitemia after a chloroquine regimen (25 mg/kg of body weight) was commonly observed. Persistent P. falciparum parasitemia was recorded in 8% cases. Poor response to chloroquine suggests the need to change the drug policy. PMID- 9740262 TI - Indirect inhibition by antibiotics of nucleotide and deoxynucleotide biosynthesis in Plasmodium falciparum. AB - The effects of the antibiotics, doxycycline, azithromycin, ciprofloxacin and chloramphenicol, upon levels of nucleoside-5'-triphosphates (NTPs) and 2' deoxynucleoside-5'-triphosphates (dNTPs) have been compared in the malarial parasite, Plasmodium falciparum, and in human CCRF-CEM leukemia cells. All 4 antibiotics had more severe effects upon levels of NTPs and dNTPs in P. falciparum compared with leukemia cells providing an explanation for their selective toxicity against malaria and their utility as antimalarial drugs. In bacteria, the first 3 drugs inhibit protein synthesis while ciprofloxacin inhibits topoisomerase II. The observed depletions of NTPs and dNTPs would be a secondary effect of the drug but may result in death of the parasite. PMID- 9740261 TI - Pharmacokinetic interactions of artemether and pyrimethamine in healthy male Thais. AB - The pharmacokinetics of a single oral dose of artemether (300 mg) and pyrimethamine (100 mg) given as each individual drug alone or as a drug combination (artemether 300 mg plus pyrimethamine 100 mg), were investigated in 8 healthy male Thai volunteers. Both artemether and pyrimethamine were rapidly absorbed after oral administration. Elimination of pyrimethamine was however, a relatively slow process compared with artemether, and thus resulted in a long terminal phase elimination half-life (50-106 hours). Pharmacokinetics of artemether and dihydroartemisinin following a single oral dose of artemether alone or in combination with pyrimethamine were similar. In contrast, coadministration of artemether resulted in significantly increased Cmax (medians of 818 vs 1,180 ng/ml) and contracted the apparent volume of distribution (medians of 3 vs 2.56 l/kg) of pyrimethamine. PMID- 9740263 TI - The occurrence of dihydrofolate reductase (DHFR) point mutation (SER-108-->ASN 108) in Malaysian isolates of Plasmodium falciparum. PMID- 9740264 TI - Opportunistic protozoa in stool samples from HIV-infected patients. AB - A retrospective study of stool samples of HIV-infected patients from January 1994 to December 1995 submitted to the Department of Tropical Pathology was analyzed. There were twenty-two cases, all of which presented with chronic diarrhea. Result showed that 50% were infected with protozoa. These include Microsporidium (27.27%), Cryptosporidium (9.09%), Isospora belli (4.54%) and Giardia intestinalis cysts (9.09%). Other infections were Candida sp, Strongyloides stercoralis larva and Opisthorchis viverrini ova. The data stress the importance of opportunistic protozoa in the HIV-infected patients. Awareness of their existence of the diseases is important areas with increasing number of HIV infected patients for early detection and proper treatment. PMID- 9740265 TI - Diagnosis of Plasmodium falciparum infection using ParaSight(R)-F test in blood and urine of Papua New Guinean children. AB - Rapid, simple, accurate and cheap methods are required for the diagnosis of malaria in endemic areas. The ParaSight(R)-F test, which is based on qualitative detection by monoclonal antibody of the Plasmodium falciparum (Pf) histidin-rich protein-II in the blood, showed promising results. As some antigens of Pf have been detected in the urine, we assessed the efficiency of the ParaSight(R)-F test in the whole blood and in the urine using microscopy and PCR as gold standards. One hundred and twelve children living in the East Sepik Province of Papua New Guinea (PNG) were recruited during a cross-sectional community survey. When using microscopy as reference, the ParaSight(R)-F test applied to whole blood had 84% sensitivity and 77% specificity. The semi-quantitative assessment showed that the intensity of the color on the wick correlated with parasite density. The ParaSight(R)-F test performed on urine had 81% sensitivity but only 26% specificity. Very similar results for blood and urine were obtained when using PCR as reference. The present evaluation of the ParaSight(R)-F test applied to blood compares well with findings in endemic areas of Africa or Asia, and confirms its usefulness to diagnose Pf infection in endemic areas of the South Pacific. Because of the lack of specificity, the ParaSight(R)-F test performed on urine cannot be recommended. PMID- 9740266 TI - Lack of significant association between rosette formation and parasitized erythrocyte adherence to purified CD36. AB - The ability of Plasmodium falciparum infected erythrocytes from 162 Thai patients with uncomplicated malaria, 82 patients with severe malaria and 19 patients with cerebral malaria to form rosettes in vitro was studied. Of 263 isolates, 62 were evaluated for their adherence to different target molecules. We found that wide variation occurred in isolates from all groups in the level of rosette formation and adherence to CD36, intracellular adhesion molecule-1, thrombospondin and chondroitin sulfate A. No statistically significant correlation between the magnitude of rosette formation and disease severity was found (p > 0.05). In addition, our results from the use of purified CD36 as an adherence receptor showed no association between the degree rosetting and level of cytoadherence (p > 0.05, r = -0.04). Our data provide evidence that rosette formation and cytoadherence involve different molecular mechanisms and both phenomena can occur in all manifestations of the disease. PMID- 9740267 TI - Serum cortisol levels in patients with uncomplicated and cerebral malaria. AB - Ten patients with uncomplicated malaria, ten with cerebral malaria and 37 controls (blood donors from blood bank) were included in the study. The serum cortisol levels of the patients were determined daily for 7 days while they were at the hospital. A radio-immunoassay method was used for quantitative measurement of cortisol in human serum. The mean serum cortisol level of patients with uncomplicated malaria was 528.2 +/- 123.9 nmol/l, with cerebral malaria was 516.0 +/- 80.5 nmol/l, and in controls was 393.8 +/- 141.0 nmol/l. There was a significant rise of serum cortisol levels in patients with malaria when compared to controls at the day of admission to hospital. There was no significant difference between uncomplicated malaria patients and those with cerebral malaria. There was also no significant difference between the different days of treatment up till day 7. We found no cortisol insufficiency in cases with falciparum malaria during acute and convalescent stages of illness. PMID- 9740268 TI - Nutritional status of school children in an endemic area of iodine deficiency disorders (IDD) after one year of iodine supplementation. AB - To improve the health and nutritional status of school children in an area of iodine deficiency disorders (IDD) by means of different iodine fortifications in salt, fish sauce and drinking water, anthropometric assessment for nutritional measurement, including hematological status, were performed. There was a significant difference in the weight and height of the children from the four schools investigated, before and after supplementation in each school. The prevalence of anemia (as indicated by hematological measurement) and iodine deficiency (as indicated by urinary iodine concentration in the children from the four schools) were assessed and compared before and after iodine supplementation; a decrease in prevalence was found in all school children, however, serum ferritin did not change before and after supplementation. PMID- 9740269 TI - Health status of Orang Asli (aborigine) community in Pos Piah, Sungai Siput, Perak, Malaysia. AB - A study of health status of Orang Asli population (based on physical examination findings) was conducted in 4 villages in Pos Piah, Sungai Siput Perak, Malaysia. In all 356 individuals between 4 months-72 years old (178 males and 178 females) participated in this study. Poor general health status, physical and mental handicaps were seen in 7.8%, 0.3% and 0.3% of the population, respectively. About one-fifth of the population had dental caries. Splenomegaly, hepatomegaly and hepatosplenomegaly were among the commonest abnormalities with the occurrence rates of 19.8%, 13.7% and 6.7%, respectively, being detected in the population. About one-fifth of the population showed signs suggestive of protein-energy deficiency; whilst less than 5% showed signs indicative of riboflavin, iodine and iron deficiencies. Vitamin A deficiency was the commonest nutritional deficiency identified in this community with almost 38.4% of them showing signs of the deficiency. The commonest skin infection was scabies. PMID- 9740270 TI - Preliminary report on the short stature of Southeast Asian forest dwellers, the Manni, in southern Thailand: lack of an adolescent spurt in plasma IGF-I concentration. AB - Plasma concentration of insulin-like growth factor type-I (IGF-I) was studied among the Mannis in Thailand to find a possible cause of their short stature. The Mannis are hunting and gathering indigenous tribal peoples living in Asian tropical rain forests. A total of 50 plasma specimens from three different Manni groups in southern Thailand were used in this study. The concentrations of acid ethanol extract of plasma IGF-I were measured by radio-immunoassay. We found that (1) plasma concentration of IGF-I in the Mannis was low, (2) there was no adolescent spurt in IGF-I levels, and (3) the post adolescent plasma IGF-I level of the Manni was significantly lower than that of age-matched Japanese. Low IGF-I levels among the Mannis may account for their short stature. PMID- 9740271 TI - Tobacco use in Mizoram, India: sociodemographic differences in pattern. AB - A study on tobacco use was carried out in Aizawl district of Mizoram, India, to assess the prevalence and pattern of tobacco use. An area served by two Sub health Centers representing town and village population were selected for a household survey. 375 people (age 10 years and above) were interviewed about their habits of taking tobacco. Use of tobacco was high among males (56.6%) and females (45.7%), but the high prevalence of smoking among males (42.3%) and chewing among females (27.9%) indicates the existence of sex differences in tobacco use pattern. Age and occupation had significant association with tobacco use but influence of education was very low and its association was not significant. Mean age for start of tobacco chewing and smoking for males and females varied significantly. However, the mean age of start for adolescent and young age (10-29 years) tobacco users was 17.2 years (SD +/- 2.3). Though there are some limitations to this study, these findings revealed differential patterns of tobacco use which is valuable information for prevention effort. PMID- 9740272 TI - K-ras oncogene and p53 gene mutations in cholangiocarcinoma from Thai patients. AB - Paraffin embedded tissues from twenty Thai patients with intrahepatic cholangiocarcinomas were studied for K-ras gene mutations at codon 12, 13 and 61 and for p53 gene mutations in exon 5 to 8 using polymerase chain reaction and thermal cycle sequencing. Results showed that point mutations at these regions in K-ras oncogene were not present in all the samples. One case harbored a p53 gene mutation in codon 282 in exon 8, CGG (arginine) to TGG (tryptophan), but the mutation was not found in other patient's tissues with similar histological features. PMID- 9740273 TI - Non-cirrhotic portal fibrosis associated with pulmonary arteriovenous communication and pulmonary arterial hypertension. AB - A case of non-cirrhotic portal fibrosis associated with pulmonary arteriovenous communication and pulmonary arterial hypertension is reported. The patient was a 7-year old boy who presented with hematemesis, cyanosis, hypoxemia and orthodeoxia. His liver pathology was compatible with non-cirrhotic portal fibrosis. His pulmonary angiography showed arteriovenous shunting and pulmonary arterial hypertension (mean pulmonary artery pressure 34 mmHg). His sister also had non-cirrhotic portal fibrosis with neither hypoxemia nor orthodeoxia. This report raises the possibility of non-cirrhotic portal fibrosis having a genetic etiology. PMID- 9740274 TI - Prevalence of North India of hepatitis B carrier state amongst pregnant women. AB - The study was undertaken to determine the hepatitis B carrier rate in North India along with the relative infectivity of the carriers. A total of 1,112 pregnant women were investigated for hepatitis B carrier state during their routine visits to antenatal clinics. All three tiers of the health care delivery system were included from four regions of North India. The sera were screened for the presence of hepatitis B surface antigen (HBsAg), hepatitis B "e" antigen (HBeAg), and antibody to hepatitis B "e" antigen (Anti-HBe) by third generation Macro ELISA tests. The average hepatitis B surface antigen carrier rate was 9.5%. The carriers were found to be of relatively low infectivity with HBeAg and Anti-HBe present in 12.0% and 25.3% of the HBsAg carriers respectively, and both these markers absent in 62.7%. It was concluded that in the past decade the hepatitis B endemicity in North India has probably increased, but the relative infectivity of the carriers remains the same. PMID- 9740275 TI - Primers for Salmonella serovar detection by polymerase chain reaction. AB - Salmonella serovar detection was studied by polymerase chain reaction (PCR). The primers were designed from Salmonella specific clone, A18:2 which was previously constructed and studied for genus specificity through colony hybridization. The primers were subsequently tested for specificity and sensitivity and showed that they amplified DNA fragment of all Salmonellae tested but did not amplify all isolates of non-Salmonellae tested. The amplified fragment was confirmed and increased sensitivity by nested PCR. Salmonella isolates amplified by the primers in the first round PCR were all positive in the second round. The sensitivity in the first and second round were 7 pg and 80 fg, respectively. The result indicated that the primers can be used as molecular tool for future field survey of Salmonella both in food and in clinical specimens. PMID- 9740277 TI - Serological findings of Coxiella burnetii infection among patients with fevers in a health centre in Sarawak, Malaysia. PMID- 9740276 TI - Enteropathogenic Escherichia coli in raw and cooked food. AB - A total of 402 Escherichia coli isolates were obtained from a variety of food samples and screened for enteropathogenic E. coli (EPEC). Screening was carried out using 15 specific monovalent antisera from Murex Diagnostic Limited. A total of 19 E. coli isolates were serotyped as EPEC. The EPEC strains were shown to belong to 8 serotypes. Eight out of 19 EPEC strains belonged to serotype 018C:K77 (B21). Seventeen out of 19 of the EPEC strains were isolated from cooked food. The presence of E. coli in cooked food is an indicator of fecal contamination and a sign of unhygienic food handling. The presence of EPEC in food could be a potential source of food-borne outbreak. Hygiene training for every food-handler is a necessity. PMID- 9740278 TI - Bacterial pathogens (non-Mycobacterium) from sputum culture and antimicrobial susceptibility. AB - Sputum culture of patients at Siriraj Hospital, Bangkok was 49.84% positive for bacterial pathogens in 1994 and 40.95% in 1995. The average incidence of gram negative rods was 3.11 fold more than the combination of gram-positive cocci and gram-negative cocci. The most common gram-negative rod was Pseudomonas aeruginosa, followed by either Klebsiella pneumoniae or Acinetobacter anitratus depending on year. The most common coccus was Staphylococcus aureus. From both years, the number of Haemophilus influenzae, Streptococcus pneumoniae, Burkholderia pseudomallei and Nocardia spp isolated were 122, 93, 13 and 11 strains respectively. For antimicrobial susceptibility, P. aeruginosa was sensitive to ceftazidime, imipenem, gentamicin, amikacin, netilmicin, ciprofloxacin (range 56-89%). S. aureus (MSSA) was sensitive to common used drugs. S. aureus (MRSA) was sensitive to co-trimoxazole, fosfomycin, vancomycin (range 57-100%) and resistant to most drugs. PMID- 9740279 TI - Cryptosporidium infection in HIV-seropositive and seronegative populations in southern Thailand. AB - The prevalence of Cryptosporidium infection in 61 HIV-seropositive and 61 HIV seronegative subjects (aged less than one to 67-year-old) in Songkhla City, southern Thailand was studied by a centrifugal flotation technic using sucrose solution. Most of the HIV-seropositive subjects (72%) were 20 to 39 years old. Cryptosporidium oocysts were detected in 10% (6/61) of HIV-seropositive and in 2% (1/61) of HIV-seronegative subjects. Infection rates in these two groups, however, were not statistically significant (p > 0.05). The number of Cryptosporidium oocysts observed in 20 microscopic fields ranged between one and over 12,000. Among the seven Cryptosporidium-positive subjects, six were adults (18 to 42-year-old) and one was three-year-old child. All of the Cryptosporidium infected subjects were male, and two of them were passing formed (normal) feces. Biochemical findings revealed dishepatica in five of six Cryptosporidium infected HIV-seropositive subjects. PMID- 9740280 TI - Massive pulmonary cryptococcosis in an immunocompetent patient. AB - A 64-year-old man presented with progressive dyspnea. The symptom of severe hypoxia requiring mechanical ventilator, and bilateral pulmonary infiltrates on the chest film led to the clinical diagnosis of adult respiratory distress syndrome. Autopsy demonstrated widespread cryptococci and mucinous material in alveoli with mild inflammatory response. PMID- 9740282 TI - Intestinal geohelminthiasis and growth in pre-adolescent primary school children in Northeastern Peninsular Malaysia. AB - A cohort of one hundred 8-9 year old school children in Northeastern Peninsular Malaysia underwent stool examination, weight and height measurements. Seventy three children were infected with Ascaris lumbricoides and/or Trichuris trichiura. All infected children were treated with albendazole at baseline, 6 months and 9 months. Measurements were repeated on all but 2 children at 1 year. Repeat stool examination (n = 94) at 1 year revealed a marked reduction in the level of Ascaris infection and a modest reduction in Trichuris infection. There was no difference in net growth between treated children and uninfected controls. Post-hoc analysis by gender however revealed that infected girls (n = 33) experienced significantly higher increments in weight, height and weight for age. Furthermore, children found to be infected at baseline level but worm free at follow-up, were observed to have experienced greater increments in height and height for age. The evidence suggests that periodic antihelminthic treatment may have a positive effect on the growth of subsets of pre-adolescent children but it is emphasised that further work is required to validate these findings. PMID- 9740281 TI - Comparison of the efficacy of tetracycline and norfloxacin in the treatment of acute severe watery diarrhea. AB - Antibiotic treatment appears to shorten the duration of diarrhea and eradicate Vibrio cholerae. The objective of this study was to compare the efficacy of tetracycline with norfloxacin therapy in patients (adults and children) with acute severe watery diarrhea caused by VC 01 and VC 0139. Patients (adults and children) with acute severe watery diarrhea admitted to Bamrasnaradura Infectious Disease Hospital, Thailand were randomized to receive either tetracycline (500 mg qid in adults and 12.5 mg/kg qid in children) or norfloxacin (400 mg bid in adults and 7.5 mg/kg bid in children) for 3 days each. The duration of diarrhea and the fecal shedding were comparable between two groups. Thirteen cases were treated with tetracycline and twelve cases with norfloxacin. The results showed the mean duration of diarrhea in tetracycline-treated and norfloxacin-treated groups were 1.31 and 1.25 days, respectively. The mean fecal shedding in tetracycline-treated and norfloxacin-treated group were 1.38 and 1.33 days, respectively. However, there were no statistically significant differences between two groups of both comparisons (p > 0.05). All isolates (VC 01 and VC 0139) in this study were susceptible to both antibiotics. Tetracycline therapy is as good as norfloxacin therapy for quick recovery and time for bacterial eradication in patients with acute severe watery diarrhea caused by Vibrio cholerae. Children aged less than 8 years should not use tetracycline therapy because of its toxic effects. PMID- 9740283 TI - Morphological variation and abnormality of cephalic hooklets of Gnathostoma spinigerum hepatic stage larvae from laboratory infected mice. AB - One thousand advanced third-stage larvae of Gnathostoma spinigerum from laboratory infected mice, two to five weeks after being fed with infected cyclops, were examined specifically for the morphology of their cephalic hooklets. Among these, only the 15-day old (early hepatic-stage) larvae and the 30-day old (late hepatic-stage) larvae were measured for the size of their body and hooklets. The average body size of the 15-day old and 30-day old larvae were 3.4 +/- 0.4 x 0.4 +/- 0.04 mm and 4.9 +/- 0.4 x 0.5 +/- 0.04 mm, respectively. The average size of the hooklets from rows one to four of the 15-day old larvae was 14.6 +/- 1.7 x 6.8 +/- 0.6 microm, 15.6 +/- 2.0 x 7.2 +/- 0.5 microm, 16.0 +/ 1.8 x 7.4 +/- 0.6 microm and 15.9 +/- 1.9 x 7.3 +/- 0.6 microm, respectively. Those of the 30-day old larvae were 15.1 +/- 1.7 x 7.1 +/- 0.6 microm, 16.3 +/- 1.6 x 7.5 +/- 0.7 microm, 16.5 +/- 1.7 x 7.8 +/- 0.6 microm and 16.3 +/- 1.7 x 7.6 +/- 0.8 microm, respectively. The average number of cephalic hooklets from rows one to four of the two- to five-week old larvae were 42.8 +/- 2.6, 45.3 +/- 2.8, 46.9 +/- 2.8 and 50.2 +/- 2.9, respectively. Several types of morphological variation and abnormality of the cephalic hooklets were observed. The most common ones were extra rudimentary hooklets below row four or within the four rows of hooklets (10.8%), the present of a fifth row of hooklets (1.9%), abnormal hooklets in only row four (1.2%), lobed or branched hooklets (0.5%), spiral arrangement of the four rows of hooklets (0.4%), and fragmented hooklets (0.4%). PMID- 9740284 TI - Opisthorchiasis and intestinal fluke infections in northern Thailand. AB - Four hundred and thirty-one residents from 16 provinces in northern Thailand who had previously been found positive for Opisthorchis viverrini or Opisthorchis viverrini-like eggs were given praziquantel 40 mg/kg. The stool was collected for 4 to 6 times and examined for adult worms. The prevalence of Opisthorchis viverrini in this group was 11.6%. Intestinal flukes, Haplorchis taichui and Haplorchis yokogawai, were predominantly found in 63.11% and 10.44% respectively. Other intestinal flukes (Centrocestus caninus, Echinostoma malayanum, Haplorchis pumilio, Phaneropsolus bonnei, Plagiorchid flukes, Prosthodendrium molenkampi and Stellantchasmus falcatus) were also found in small numbers. PMID- 9740285 TI - A new technic for counting Schistosoma japonicum eggs in pig feces. AB - An improved laboratory method was developed for counting Schistosoma japonicum eggs in pig feces, which involves filtration, sedimentation and centrifugation, but avoids toxic chemicals. It is sensitive, allows easy differentiation from similar-sized and shaped protozoan cysts, and permits evaluation of egg viability both by direct viewing of eggs and miracidial hatching. It was found to be significantly better at recovering eggs than the modified Bell filtration technic. The sensitivity, specificity and practicality of this technic make it our method of choice for studies on porcine schistosomiasis japonica. PMID- 9740286 TI - Cutaneous leishmaniasis in Nepal. AB - We report an imported case of cutaneous leishmaniasis in a 30 year old adult male from Nepal caused by Leishmania tropica. This case from Dharan is the first such report of imported cutaneous leishmaniasis in Nepal. PMID- 9740287 TI - A first record from Thailand of human external ophthalmomyiasis due to Oestrus ovis. AB - A case of external ophthalmomyiasis caused by Oestrus ovis was first reported from Lopburi Province, Central part of Thailand, in January, 1997. A 18-year-old man presented with a several hours history of foreign body sensation in his left eye accompanied by irritation. Eight Oestrus ovis first-instar larvae were removed from lower palpebral conjunctiva. Symptoms and clinical signs resolved in 24 hours after mechanical removal of the larvae. There was no history of contact fly and domestic animals. The larvae were identified by light microscopic examination. PMID- 9740288 TI - Transmission dynamics of filariasis in Khurdha district of Orissa, India. AB - A three year longitudinal study was carried out to quantify the different parameters of filarial transmission in an endemic area of Orissa State, India. Parasitological surveys revealed mean microfilaria rate, microfilaria density and median microfilaria density (MFD-50) to be 9.41, 19.23 and 7.33, respectively. The per man hour density of the vector, Culex quinquefasciatus varied from 24.2 to 66.0 with a peak in January. Infection rate varied from 0.9 to 27.5%, while infectivity rate ranged between 0.0 and 15.2%. Infectivity rate showed high correlation with microfilaria rate and per man hour density of adult mosquito. The highest numbers of first stage larvae (L1), second stage larvae (L2) and third stage larvae (L3) per mosquito were found to be 25, 22 and 11, respectively. Average L3 load per infective mosquito ranged from 1.0 to 7.2. L3 load showed high correlation with microfilaria rate (r = 0.845, p < 0.01) while no correlation was seen with microfilaria density. PMID- 9740289 TI - Possible transmission of two types of Wuchereria bancrofti in Muang District, Chiang Mai, northern Thailand. PMID- 9740290 TI - Gnathostome infection in swamp eels, Fluta alba, in central Thailand. AB - To investigate the distribution of gnathostome worms in central Thailand, the infective larvae of Gnathostoma spp were examined from the flesh and liver of swamp eels, Fluta alba. Seven hundred and eighty-eight eels were purchased from markets in 11 provinces; Ang Thong (30), Ayutthaya (36), Chachoengsao (30), Lop Buri (30), Nakhon Nayok (437), Pathum Thani (30), Prachin Buri (48), Ratchaburi (53), Saraburi (30), Samut Prakan (30) and Suphan Buri (34). The highest rate of gnathostome infection was observed in swamp eels from Nakhon Nayok (68.7%). The infection rates in Ayutthaya, Ang Thong, Prachin Buri, Ratchaburi, Saraburi and Lop Buri were 33.3%, 26.7%, 25.0%, 18.9%, 13.3% and 10.0% respectively. Gnathostome larvae were not found in swamp eels from Chachoengsao, Pathum Thani, Samut Prakan and Suphan Buri. Among the 9,573 larvae recovered, almost all were the advanced third stage larvae of G. spinigerum, except one larva from Nakhon Nayok and two larvae from Ratchaburi which were identified as the advanced third stage larvae of G. vietnamicum and G. hispidum respectively. This study is the first report of swamp eels as natural intermediate hosts of G. vietnamicum and G. hispidum. PMID- 9740291 TI - Seasonal variation in the intensity of Gnathostoma larvae in swamp eels (Fluta alba) sold in a local market in Bangkok. AB - The viscera of swamp eels were obtained from a local market in Bangkok twice a month from June 1996 to May 1997. The livers were separated, weighed and counted. Gnathostome larvae were recovered from the livers by the digestion technic, examined, identified, and counted. A total of 12,278 Gnathostoma larvae were obtained from 18,561.1 g (15,264 pieces) of eel livers. The overall average number of larvae/g liver and the overall average number of larvae/liver are 0.91 and 0.94, respectively. The greatest number of larvae/g liver (on average) was in December (high levels of infection during the months of October to December) whereas the lowest was in April (lowest levels of infection during the months of March to April). Thus there was a marked decrease in the average number of larvae/g liver during January to April, which then started to rise in May. This finding suggests that the level of infection abruptly decreases soon after the completion of the rainy season, starts to rise when the rain has come, and reaches its peak when the amount of rainfall is highest. More than 99% of the total gnathostome larvae recovered were identified to be G. spinigerum, and 25.4% of the entire larvae recovered bore variant or abnormal cephalic hooklets. The most common unusual feature was that there were extra rudimentary hooklets above row one, below row four and in between the four rows of hooklets which comprised 21.4%. In addition, the body size and the number of cephalic hooklets of G. spinigerum are also discussed. PMID- 9740292 TI - A thirty day course of sodium stibogluconate for treatment of Kala-azar in Nepal. AB - Twenty-seven cases of Kala-azar were treated with sodium stibogluconate at a dose of 20 mg/kg/day for 20 days (group A) and an equal number of cases were treated with the same dose but for a longer duration of 30 days (group B). Clinical and laboratory evaluation of these cases were carried out before and after therapy, during a follow up of cases every month, upto 6 months. Renal and liver function tests and electrocardiography were carried out of monitor any toxic effect of the drug during therapy. The cure rates of patients were 77.78% and 92.59% in group A and B cases respectively. Six and two patients in group A and B respectively were unresponsive to the treatment and showed relapse. Results of the study show that treatment of cases of Kala-azar with sodium stibogluconate in a dosage of 20 mg/kg/day for a longer period of 30 days is effective with a higher cure rate and minimum side effects, for treatment of cases of Kala-azar in this eastern part of Nepal, endemic for the disease. PMID- 9740294 TI - Characteristics of malaria vector breeding habitats in Sri Lanka: relevance for environmental management. AB - In and around a village in the Anuradhapura District of Sri Lanka anopheline larvae were sampled from July 1994 to April 1996 in all surface water bodies. Samples positive for Anopheles culicifacies, the established vector of malaria in Sri Lanka, and for An. barbirostris, An. vagus, and An. varuna, potential secondary vectors, were characterized by site, exposure to sunlight, substratum, turbidity of the water, presence of vegetation, and presence of fauna. Availability of pools of stagnant water in the stream near the village and along the edge of the village tank was highly predictive for presence of An. culicifacies larvae, independent from the other characteristics that were included in the study. The biological and physical characteristics could not very well explain the preference for certain habitats, but it was of interest that An. culicifacies, generally considered to bread in sun exposed clear water pools, was able to exploit habitats that were shaded and contained turbid water. Environmental management interventions to control An. culicifacies breeding have to take into account that the secondary vectors of malaria exploit other habitats and would not be affected by the interventions. PMID- 9740293 TI - Comparative susceptibility of two forms of Anopheles sinensis Wiedemann 1828 (Diptera : Culicidae) to infection with Plasmodium falciparum, P. vivax, P. yoelii and the determination of misleading factor for sporozoite identification. AB - Two karyotypic forms of laboratory-raised Anopheles sinensis, ie Form A (XY1) and Form B (XY2), were experimentally infected with various indigenous strains of Plasmodium falciparum and P. vivax using an artificial membrane feeding technique, and a rodent malaria, P. yoelii, using a direct feeding technic and dissected 7-9 days and 10-15 days after feeding for oocyst and sporozoite rates, respectively. The results revealed that two forms of An. sinensis were refractory vectors for P. falciparum and P. yoelii since 0% of oocyst and sporozoite rates were obtained, but poor vectors for P. vivax since 0.00-85.71% and 0.00-5.88% of oocyst and sporozoite rates were recovered. The sporozoite-like crystal found in the median lobe of the salivary gland of An. sinensis which could be a misleading factor in identification of true sporozoites in the salivary glands is reported for the first time. PMID- 9740295 TI - Distribution of potential dengue vectors in major townships along the national highways and trunk roads of northeast India. AB - Surveys were conducted in some townships along the national highways and trunk roads of northeast India to detect breeding of Aedes mosquitos in used/waste tire dumps piled outdoors by the tire repairing shops during summer season of 1996 1997. The breeding of both the potential vectors of dengue, viz. Aedes aegypti and Ae. albopictus were detected, prevalence rate being in the range of 30.0-88.0 (CI = container index value). The preponderance of Ae. aegypti was considerably much higher than that of Ae. albopictus and all the urban and semiurban areas coming up along the side of the roads were observed to be infested with Ae. aegypti. With respect to transmission of dengue, this study clearly indicates that waste tire dumps in every urban agglomeration should receive primary attention in view of their relative contribution to the abundance and dispersal of these vector mosquitos. PMID- 9740296 TI - Estimation of predation by the larvae of Toxorhynchites splendens on the aquatic stages of Aedes aegypti. AB - Predation by instars of Toxorhynchites splendens on aquatic stages of Aedes aegypti was studied by estimating functional response parameters such as attack rate (a') and handling time (Th) in the laboratory. The predator displayed typical type-II functional response, similar to that of most insect predators when presented with increasing densities of any given size class of prey. Second instar predator attacked prey significantly at higher rate than the other instars. Small prey were attacked at higher rate than the predation on larger prey. Except second instar predator, other instars showed significant reduction in a' with increase in Th. Foraging surface area did not influence the predation rate. Predation was high at high water temperature and this was more prominent in the second instar predator. However, prey handling time was independent of the water temperature. Modeling of the predation of mixed age populations of prey and the predator through this short-term functional response experiment is discussed. PMID- 9740297 TI - Deposits of different origin in the lungs of the 5,300-year-old Tyrolean Iceman. AB - Deposits in the lung of the Late Neolithic Tyrolean Iceman were studied with a combination of different methods of analytical electron microscopy. Numerous anthracotic areas with plentiful inhaled soot particles were found in the lung; these most probably derived from open fires in houses. Between the soot particles tiny mineral crystals (mainly muscovite) were identified, which may indicate that the Tyrolean Iceman lived in a muscoviterich area. Furthermore, illite, quartz, and a plagioclase (andesine), which are also minerals in the crystalline rocks of the Otztal Alps, were found. Additionally, organic material, which may represent inhaled threshing residues, was present in the anthracotic areas. As threshing residues and seeds in husk also were detected in the Iceman's belongings, some kind of rustic occupation seems probable. Outside of the anthracotic areas, vivianite and hydroxyapatite crystals were detected. Because of their separate location, and as vivianite is also described in the Iceman's skin, these minerals seem to have crystallized during his 5,300 years of storage in the high mountains. PMID- 9740299 TI - Dental defects of congenital syphilis. AB - Diagnosis of the congenital form of syphilis is an important part of the palaeopathology of this disease. In theory, there are clear clinical signs to be found in the long bones and teeth, but it has rarely been possible to recognise the latter with a confidence in archaeological material, partly because the original descriptions of the dental deformities are sometimes contradictory and partly because it is nowadays difficult to find reference specimens in museums. This article describes two such specimens which have recently been rediscovered, and discusses the form of the dental defects which they show (Hutchinson's incisors, Moon's molars, and mulberry molars) in relation to the developmental sequence of the teeth. PMID- 9740298 TI - Methods for improving the efficiency of estimating total osteon density in the human anterior mid-diaphyseal femur. AB - In order to preserve whole bone integrity and minimize destruction, paleohistologists often rely on histomorphometric data obtained from small areas (1.5-50 mm2) sampled within the anterior mid-diaphyseal femur. Because bone exhibits significant histological variation, the validity of results based on such sampling is questionable. The accuracy of various subareas (columns, rows, squares approximating dimensions and locations assessed by paleohistologists) in predicting total osteon density in the anterior mid-diaphyseal femur is assessed in the present study. Thirty-five specimens (12.7 mm wide, 100 microm thick, average area 56.7 mm2) were chosen at random from a skeletal population of 94 Inuits and Pueblo agriculturists. The specimens were photographed and enlarged; an acetate grid (12 columns, 10 rows, 120 squares, square = 1 mm2 of bone surface) was superimposed over the photograph; and secondary osteons and fragments were identified. Alternate columns (50% total area, T.Ar) predicted over 98% of entire section total osteon density. Two column combinations (15% T.Ar), separated by at least one column, predicted 91 to 95% of total osteon density. Individual column (8% T.Ar) predictability ranged from 48 to 86%. Two row combination (32 to 40% T.Ar) predictability values ranged from 86 to 95%. Individual rows (<1 to 20% T.Ar) predicted from 45 to 92% of total variation. Combinations of squares approximating areas and locations assessed by other paleohistologists ranged in predictability values from 80 to 94%. The results demonstrate that subareas of as little as 15% predict 95% of variation in total osteon density in the entire anterior mid-diaphyseal femoral section. A minimization of histological area evaluated without the loss of accuracy allows for a minimization of time invested in data collection and the utilization of partially damaged specimens. PMID- 9740300 TI - Rare temporal bone pathology of the Singa calvaria from Sudan. AB - Evidence has recently accumulated that the Singa calvaria from Sudan probably dates from Oxygen Isotope Stage 6 (>130 ka). Morphological studies have indicated a mixture of archaic and more modern human traits, but such analyses are complicated by the possibility that the vault is pathologically deformed, although the exact etiology has not been established. Now computed tomography (CT) has revealed that the right temporal bone lacks the structures of the bony labyrinth. The most likely cause of this rare pathological condition appears to be labyrinthine ossification, in which newly deposited bone obliterates the inner ear spaces following an infectious disease or occlusion of the labyrinthine blood supply. A possible cause of vascular compromise could have been the presence of an expanding acoustic neuroma in the internal acoustic meatus, which is suggested by a significantly wider right meatus compared with the left side. Interestingly, labyrinthine ossification is also consistent with the controversial diagnosis that an anemia caused the characteristic diploic widening at the parietal bosses, because prime etiological factors of ossification are among the common complications of some of these blood diseases. CT examination of the vault and a review of the literature suggest that a blood disorder may well have caused the unusual parietal morphology. Given the nature of these pathological conditions, the Singa individual must have experienced a period of considerable disability. The morphological evidence from the normal bony labyrinth on the left side and from the CT evaluation of the vault is consistent with the interpretation of Singa as a late archaic hominid or an early representative of Homo sapiens drawn from a population which might be directly ancestral to modern humans. PMID- 9740301 TI - Skeletal evidence of health and disease in pre-contact Alaskan Eskimos and Aleuts. AB - There have been relatively few paleopathological studies of arctic populations to date, compared to other regions of North America. Studies aimed at elucidating patterns of health and disease in arctic peoples prior to contact and assessing inter- and intraregional differences in disease patterns have been particularly few. In the present study, five pre-contact skeletal samples (N = 193), representing 4 Eskimo populations from northern coastal Alaska and 1 Aleut population from the eastern Aleutian Islands, were examined macroscopically for the following indicators of health status: cribra orbitalia, porotic hyperostosis, trauma, infection, dental caries, abscesses, antemortem tooth loss, periodontal disease, and dental attrition. In addition, archeological and epidemiological data were used to help reconstruct the health of these populations. The goals of the analysis were 2-fold: 1) to assess the pre-contact health of North Alaskan Eskimos and Aleuts in order to provide a baseline comparison for the post-contact health of these groups, and 2) to determine if any differences in disease patterns exist between the Eskimos and Aleuts that might be related to differences in their physical environment, subsistence patterns, and cultural practices. The analysis revealed that both groups suffered from a variety of health problems prior to contact, including iron deficiency anemia, trauma, infection, and various forms of dental pathology. Statistical comparisons of the 2 groups revealed that Eskimos and Aleuts had different patterns of health and disease prior to contact. Most notably, the Aleuts had a significantly higher frequency of cranial trauma and infracranial infection than the Eskimos, while the latter had a significantly higher frequency of enamel hypoplasia. An examination of the physical and cultural environment of the 2 groups reveals several possible explanations for these differences, including warfare, subsistence pursuits, and housing practices. The documentation of these differences indicates that variability in pre-contact disease patterns can be identified between hunter-gatherer populations living in similar environments and exhibiting similar general lifestyles. PMID- 9740302 TI - The costs of human locomotion: maternal investment in child transport. AB - This article investigates maternal investment in child carrying and presents a method for determining when it is energetically advantageous for a mother to carry her child rather than force her child to walk independently. I calculate maternal and child energy consumption while walking and develop correction factors to facilitate making these energy calculations for young children. In addition, I investigate the effect of maternal burdens in addition to the child and of external nutritional support on energy consumption. Since maternal energy is a finite resource, the "decision" to carry a child or force it to walk independently is especially important. This decision can be predicted from the body mass of the mother and child and the child's age. If the mother provides all of the child's nutrition, then the mother should choose to carry her child only when the energy usage of the mother carrying the child is less than the sum of the energy used when the mother and child walk independently. The critical velocity, when the two expenditures are equal, can then be determined. Several general hypotheses are also addressed. The critical velocity of a 60 kg mother with a 4-year-old child approximately equals the average walking speed of adult humans. For a lighter mother, the critical velocity is reached when her child is 3 years old, while for heavier mother this point is not reached until her child is 6 years old. The effect of burdens in addition to the child's mass is minimal. Nutritional support of the child by agencies other than the mother decreases the age at which the mother should force the child to walk independently. In some cases, especially for the lightest mothers, it is never in the mother's best energetic interest to carry her child. PMID- 9740303 TI - Canine tip wear in male and female anthropoids. AB - One component of the "dual selection hypothesis" (Greenfield [1992a] Year. Phys. Anthropol. 35:153-185) is that the tips of female canines are commonly blunted and more frequently so than those of conspecific males. Data derived from two randomly selected age-graded samples of Macaca fascicularis (n = 70) and Colobus badius (n = 59) show that at least 80% of the females exhibit tip blunting on one or both canines and that frequencies of blunting are far greater than those of conspecific males in both jaws. Sexual dimorphism in mandibular canine morphology and wear and other recently critiqued aspects of the "dual selection hypothesis" (Plavcan and Kelley [1996] Am. J. Phys. Anthropol. 99:379-387.) are also discussed. PMID- 9740304 TI - Use of ordinal categorical variables in skeletal assessment of sex from the cranium. AB - In anthropological studies, visual indicators of sex are traditionally scored on an ordinal categorical scale. Logistic and probit regression models are commonly used statistical tools for the analysis of ordinal categorical data. These models provide unbiased estimates of the posterior probabilities of sex conditional on observed indicators, but they do so only under certain conditions. We suggest a more general method for sexing using a multivariate cumulative probit model and examine both single indicator and multivariate indicator models on a sample of 138 crania from a Late Mississippian site in middle Tennessee. The crania were scored for five common sex indicators: superciliary arch form, chin form, size of mastoid process, shape of the supraorbital margin, and nuchal cresting. Independent assessment of sex for each individual is based on pubic indicators. The traditional logistic regressions are cumbersome because of limitations imposed by missing data. The logistic regression correctly classified 66/74 males and 46/64 females, with an overall correct classification of 81%. The cumulative probit model classified 64/74 males correctly and 51/64 females correctly for an overall correct classification rate of 83%. Finally, we apply parameters estimated from the logit and probit models to find posterior probabilities of sex assignment for 296 additional crania for which pubic indicators were absent or ambiguous. PMID- 9740305 TI - Capturing data from three-dimensional surfaces using fuzzy landmarks. AB - Anatomical landmarks are defined as biologically meaningful loci that can be unambiguously defined and repeatedly located with a high degree of accuracy and precision. The neurocranial surface is characteristically void of such loci. We define a new class of landmarks, termed fuzzy landmarks, that will allow us to represent the form of the neurocranium. A fuzzy landmark represents the position of a biological structure that is precisely delineated, but occupies an area that is larger than a single point in the observer's reference system. In this study, we present a test case in which the cranial bosses are evaluated as fuzzy landmarks. Five fuzzy landmarks (the cranial bosses) and three traditional landmarks were placed repeatedly by a single observer on three-dimensional (3D) computed tomography (CT) surface reconstructions of pediatric dry skulls and skulls of pediatric patients, and directly on four of the same dry skulls using a 3Space digitizer. Thirty landmark digitizing trials from CT scans show an average error of 1.15 mm local to each fuzzy landmark, while the average error for the last ten trials was 0.75 mm, suggesting a learning curve. Data collected with the 3Space digitizer was comparable. Measurement error of fuzzy landmarks is larger than that of traditional landmarks, but is acceptable, especially since fuzzy landmarks allow inclusion of areas that would otherwise go unsampled. The information obtained is valuable in growth studies, clinical evaluation, and volume measurements. Our method of fuzzy landmarking is not limited to cranial bosses, and can be applied to any other anatomical features with fuzzy boundaries. PMID- 9740306 TI - Fluctuating asymmetry and morphometric variation of hand bones. AB - The major aim of this study was to test three hypotheses: 1) more complex traits of the hand are less prone to developmental insults and therefore show lower fluctuating asymmetry (FA) as compared with simple traits; 2) the manifestation of FA correlates with the variability of the trait (i.e., CV); and 3) FA is an organ-wide property, and therefore a concordance exists between the FA measures of different traits in hand bones. Seventy-two bilateral measurements of hand bones, were made from plain-film radiographs of 365 cadavers. A complex trait was considered as the total length of the three phalanges of a finger and their contiguous metacarpals. Simple traits were considered to be the lengths of individual bone that made up the complex trait. The following results were obtained: 1) on the average simple traits, composing the complex trait, show much higher FA than the corresponding complex trait, but this result is expected if there is no correlation (or low correlation) between FA of simple traits within the complex trait, due to random direction of right-left differences; 2) strong and highly significant correlation was observed between FA and CV of studied traits, regardless of sex and age of individuals; and 3) the majority of FA measurements of hand bones showed no correlation. However, correlations between some sets of FA traits were highly significant. They were interpreted, although not specifically tested, as the result of a tight relationship between traits related not only developmentally but also by active performance of the same function. PMID- 9740307 TI - Technical note: Modeling primate occlusal topography using geographic information systems technology. AB - Most functional analyses of primate tooth form have been limited to linear or area measurements. Such studies have offered but a limited glimpse at differences in occlusal relief among taxa. Such differences in dental topography may relate to tooth function and, so, have considerable implications for the inference of diet from fossil teeth. In this article, we describe a technique to model and compare primate molars in three dimensions using Geographic Resources Analysis Support System (GRASS) software. We examine unworn lower second molars of three extant hominoids with known differences in diet (Gorilla gorilla, Pan troglodytes, and Pongo pygmaeus), and two fossil forms, (Afropithecus turkanesis and Dryopithecus laietanus). First, we obtained approximately 400 landmarks on the occlusal surfaces of each tooth using an electromagnetic digitizer. Raster "terrain models" of occlusal surfaces were then created by interpolation of the coordinate data. We used GRASS terrain analysis automated techniques to quantify the volumes and slopes of individual cusps. We also used the GRASS watershed technique to identify the volume of liquid that would accumulate in each tooth's basin (a measure of basin area), and the directions and intensity of drainage over the occlusal surface. In sum, GRASS shows considerable potential for the characterization and comparison of tooth surfaces. Furthermore, techniques described here are not limited to the study of teeth, but may be broadly applicable to studies of skulls, joints, and other biological structures. PMID- 9740308 TI - Seasonal effects of rotational stress on Lewis lung carcinoma metastasis and T lymphocyte subsets in mice. AB - Rotational stress specifically increases the formation of spontaneous lung metastasis in mice bearing Lewis lung carcinoma, without significantly modifying the growth of primary tumor. The increase in metastasis number and volume caused by rotational stress varies in magnitude with a highly significant circannual rhythm; the acrophase approximately coincides with summer solstice. Rotational stress causes a significant reduction in the number of CD3+ and CD4+ T-lymphocyte subsets in summer, whereas in winter the number of CD3+ subset is significantly increased; the CD4+/CD8+ ratio and the number of NK 1.1 antigen positive cells are not significantly modified by rotational stress in both periods considered. The increase in metastasis formation by rotational stress thus appears to negatively correlate with the number of splenic CD3+ and CD4+ T-lymphocyte subsets. This seasonal behavior occurs in spite of the control of light cycle, temperature and humidity in the animal housing, suggesting the existence in the host of an endogenous oscillator with a circannual period. These data indicate the opportunity to consider endogenous rhythms within the host, as well as seasonal factors, in studies on stress and neuroimmunomodulation in experimental oncology. PMID- 9740309 TI - Protein kinase C isoforms during the development of deciduomata in pseudopregnant rats. AB - In this study, we determined the expression of protein kinase C (PKC) isoforms during trauma-induced decidualization. The findings revealed that at least five PKC isoforms (alpha, delta, zeta, iota and lambda) were present in both control and decidualized tissues. After trauma-stimulation, PKC alpha was down-modulated in the deciduomata but not in the myometrium. Down-modulation was compatible with the increase in cell mitosis which reached a maximum at 2-3 days. On the other hand, PKC zeta was not down-modulated. It was increased both in the deciduomata and myometrium, and paralleled the frequency of decidual cell mitosis. The PKC isoforms of delta, iota and lambda were also increased, but they were associated with the depression of cell mitosis. Therefore, these findings suggested that the variable expression of PKC isoforms in trauma-induced decidualizing tissue in pseudopregnant rats may be involved in the modulation of decidual cell growth. PMID- 9740310 TI - Reduction of stress-induced analgesia following ethanol exposure in mice. AB - In the present study, we examined the effects of ethanol treatment on the subsequent expression of opioid and nonopioid forms of swim stress-induced analgesia (SSIA). In Experiment 1, mice were injected with ethanol (2.5 g/kg, i.p.) or an equal volume of saline once a day for two days. Animals received no treatment on day 3. On day 4, the animals were tested for opioid (3-min swim in water maintained at 32 degrees C) or nonopioid (3-min swim in water maintained at 20 degrees C) SSIA in the hotplate test (52 degrees C). Mice pretreated with ethanol injections showed a decrease in nonopioid SSIA, but not in opioid SSIA. In Experiment 2, mice were given an ethanol solution (10%) or tap water to drink for 15 days. On day 16, all animals were given tap water to drink. On day 17, the animals were tested for opioid or nonopioid SSIA. Neither form of SSIA was modified in mice that drank the ethanol solution. These results show that ethanol pretreatment can modify nonopioid endogenous analgesic responses in mice. Further, the route of administration influences the effects of ethanol pretreatment on SSIA. PMID- 9740311 TI - Rapid and sensitive determination of catecholamines and the metabolite 3-methoxy 4-hydroxyphen-ethyleneglycol using HPLC following novel extraction procedures. AB - In the present study assays were improved for the determination of free catecholamines and 3-methoxy-4-hydroxyphenethyleneglycol (MHPG), the major metabolite of peripheral and central noradrenaline. The compounds were extracted by a fluid phase extraction: a diphenyl boric acid method for the purification of catecholamines and an ethyl acetate extraction for MHPG were used, respectively. High-performance liquid chromatography with electrochemical detection was employed for quantitative analysis. In previous studies, significant differences between plasma concentrations of these substances in normal volunteers and hospital patients were demonstrated. Therefore, we established valid reference values for a hospital population. Blood and urine samples of 59 patients without disorders and medication affecting catecholamine synthesis and metabolism or the activity of the sympatho-adrenal system were collected and analyzed for free and total (free plus conjugated) MHPG, noradrenaline (NA), adrenaline (A) and dopamine (DA); total MHPG was assayed after enzymatic hydrolysis of conjugates. Our data clearly demonstrate that these methods are sensitive, specific, rapid, and can easily be standardized. The intra- and inter-assay precision were high (CV 2.6-5.3% and 4.3-6.9% for plasma, CV 3.8-4.9% and 5.1-8.2% for urine, respectively). For plasma, the mean concentrations +/- SD were determined to be 20.82+/-4.70 pmol/ml for free MHPG, 68.43+/-16.21 pmol/ml for total MHPG, 2.11+/ 0.24 pmol/ml for NA and 0.31+/-0.08 pmol/ml for A. For 24h-urine the mean concentrations +/-SD were determined to be 0.44+/-0.13 mmol/24h for free MHPG, 8.79+/-2.13 mmol/24h for total MHPG, 169.8+/-58.25 nmol/24h for NA, 62.19+/-21.79 nmol/24h for A and 757.2+/-382.6 nmol/24h for DA. In summary, these novel and rapid methods can clearly be employed in a routine clinical setting. PMID- 9740312 TI - Involvement of the pineal gland in daily scheduling of the golden spiny mouse. AB - The light-dark cycle is the major time cue for daily and seasonal scheduling of physiological activities. However, non-photic cues (e.g. environmental and social constraints) may also play a significant role. A natural model exists in the golden spiny mouse (Acomys russatus) which is nocturnal when maintained alone but diurnal when sharing a habitat with its congener, the common spiny mouse (A. cahirinus). We have recently observed that the presence of A. cahirinus provokes a major change in the daily rhythms of body temperature (Tb), and urine volume without affecting the melatonin rhythm and photoperiod-induced responses. The apparent lack of interaction between the daily and photoperiodic scheduling was further investigated by studying the significance of the pineal to the modification of A. russatus daily rhythms induced by the presence of A. cahirinus. Lesion of A. russatus pineal gland resulted in diminution of urinary 6 sulfatoxymelatonin (6-SMT) and modification of Tb and urine volume rhythms. However, the modification of Tb and urine volume rhythms provoked by the presence of A. cahirinus were similar in pineal lesioned and sham-operated A. russatus. The non-photic signals released by A. cahirinus did not significantly affect glucose utilization in the suprachiasmatic nucleus of pineal- as well as sham lesioned A. russatus. Thus, the modification of the daily scheduling of A. russatus by the photoperiod involves the pineal and/or the melatonin rhythm whereas non-photic cues effect a direct (perhaps masking), pineal-independent response to the competitor. PMID- 9740313 TI - Autoradiographic imaging of formaldehyde adducts in mice: possible relevance for vascular damage in diabetes. AB - The activity of semicarbazide-sensitive amine oxidase (SSAO) has been reported to be elevated in blood from diabetic patients. It has been suggested that the enzyme is involved in the development of complications such as retinopathies, nephropathies and neuropathies, which are associated with advanced diabetes, possibly by the formation of toxic metabolites. Under the influence of SSAO, methylamine is deaminated to formaldehyde which is known to react with various macromolecules. It has therefore been proposed that specific inhibition of SSAO could be of therapeutic value for treatment of diabetic patients. The present results provide evidence that treatment with an SSAO inhibitor potently reduces the levels of irreversible adducts. In this study, 14C-methylamine was given intraperitoneally to NMRI mice, and the tissue distribution of irreversibly bound methylamine metabolites was estimated by an autoradiographic method. Such radioactive residues occurred in high concentrations in the intestinal wall, brown adipose tissue, spleen and bone marrow. By inhibiting SSAO irreversibly with hydralazine before giving 14C-methylamine to the mice, it was possible to determine the resynthesis rate of SSAO in different tissues. A complete recovery of SSAO activity was seen in the intestinal wall after 6 days, whereas only about 60% was recovered in adipose tissue after 14 days. This suggests that factors controlling the synthesis of SSAO differ in these tissues, or that these tissues express different forms of enzymes. PMID- 9740315 TI - Neutralization of cardiac toxins oleandrin, oleandrigenin, bufalin, and cinobufotalin by digibind: monitoring the effect by measuring free digitoxin concentrations. AB - Oleandrin plant poisoning is common in children and the plant extract is used in Chinese medicines. The toxicity is due to oleandrin and the deglycosylated metabolite oleandrigenin. Bufalin and cinobufotalin (toad cardiac toxins) are also widely used in Chinese medicines like Chan SU, and Lu-Shen -WU. Severe toxicity from bufalin after consumption of toad soup has been reported. Taking advantage of structural similarities of these toxins with digitoxin, we demonstrated that these compounds can be rapidly detected in blood by the fluorescence polarization immunoassay for digitoxin. The cross reactivities of these compounds with digoxin assay were much lower. For example, when a drug free serum was supplemented with 10 microg/ml of oleandrin, we observed 127.7 ng/ml of digitoxin equivalent but only 2.4 ng/ml of digoxin equivalent concentration. Digibind neutralized all cardiac toxins studied as evidenced by significant fall of free concentrations. When aliquots of serum pool containing 50.0 microg/ml of oleandrin were supplemented with 0, 10.0, 25.0, 50.0, 100, and 200 microg/ml of digibind, the mean free concentrations were 30.6, 23.3, 16.0, 10.7, 7.8 and 5.5 microg/ml respectively. Similarly, with 50.0 microg/ml of oleandrigenin (total concentration: 36.2 ng/ml), the free concentration was 14.5 ng/ml digitoxin equivalent in the absence of digibind and 5.4 ng/ml in the presence of 200 microg/ml of digibind. In another specimen containing 500 ng/ml bufalin (total concentration: 156.9 ng/ml), the free concentration was 8.6 ng/ml in the absence of digibind and none detected in the presence of 100.0 microg/ml digibind. Because such neutralization may also occur in vivo, digibind may be useful in treating patients exposed to these toxins. PMID- 9740314 TI - Novel 21-aminosteroid U-74389G inhibits low-density lipoprotein peroxidation induced by .OH and O2-. free radicals. AB - LDL peroxidation represents one of the first event in the atherogenesis process. Inhibiting LDL oxidation may impede this process and offers a new mechanism to retard atherogenesis. 21-Aminosteroids, derived from methylprednisolone, have recently excited much interest by virtue of their ability to inhibit lipid peroxidation. The aim of our work was to investigate the effect of a novel 21 aminosteroid, U-74389G, in the LDL peroxidation initiated in a metal- and cell free system by oxygen free radicals, .OH and O2-., generated by water gamma radiolysis. In a concentration dependent manner, U-74389G increased the resistance of LDL to oxidation measured by the length of the lag phase, reduced the formation of conjugated dienes and thiobarbituric acid-reactive substances (TBARS), and also reduced the alpha-tocopherol disappearance by about 47% at the concentration 20 microM. U-74389G was also able to reduce the chemotactic activity of oxidized LDL towards monocytes, as well as the cholesterol accumulation in macrophages. These observations suggest that the U-74389G is a potent antioxidant by decreasing LDL peroxidation and this should be evaluated in in vivo models as a potential therapy to retard atherogenesis. PMID- 9740316 TI - Role of constitutive nitric oxide synthase and peroxynitrite production in a rat model of splanchnic artery occlusion shock. AB - Peroxynitrite, a potent cytotoxic oxidant formed by the reaction of nitric oxide with superoxide anion, is an important mediator of reperfusion injury. In a rodent model of mesenteric ischemia and reperfusion injury we evaluated the contribution of the constitutive and/or inducible nitric oxide synthase (cNOS or iNOS) in the formation of peroxynitrite. Splanchnic artery occlusion (SAO) shock was induced in rats by clamping both the superior mesenteric artery and the celiac trunk for 45 min, followed by release of the clamps (reperfusion). A significant peroxynitrite production was found in the plasma of the splanchnic occlusion shocked rats at 60 minutes after reperfusion. Immunohistochemical examination demonstrated a marked increase in the immunoreactivity to nitrotyrosine, a specific "footprint" of peroxynitrite, in the necrotic ileum and the aorta of shocked rats. No change in plasma levels of nitrate/nitrite, tissue iNOS expression (by western blotting detection) or iNOS activity was found in the intestine at 60 minutes after reperfusion. On the contrary, activity of the cNOS was reduced (approximately 50%) in the reperfused ischemic intestinal tissue. Treatment with NG-nitro-L-arginine methyl ester, a non selective inhibitor of nitric oxide synthase (given at 3 mg/kg i.v., 5 min prior to reperfusion), significantly reduced plasma level of peroxynitrite and the immunohistochemical staining for nitrotyrosine in the ileum and aorta. Our results suggest that during splanchnic artery occlusion shock peroxynitrite formation is likely to be correlated with nitric oxide production from constitutive nitric oxide synthase activation rather than from the inducible isoform enzyme. PMID- 9740317 TI - Oral administration of methylglyoxal leads to kidney collagen accumulation in the mouse. AB - Methylglyoxal (MG) is a physiological substrate of the glyoxalase system which is impaired in the diabetic state and implicated in the development of diabetic complications. Like other reactive aldehydes in diabetes mellitus (DM) this carbonyl can bind to and modify proteins which may lead to changes of biochemical and biophysical properties of connective tissue proteins, a hallmark of diabetes mellitus. As previous studies on MG effects were confounded by other aldehydes found in DM, we decided to administer MG to 10 healthy, female OF-1 mice for a period of five months, at a level of 50 mg/kg body weight per day using 10 healthy untreated litter mates as controls. The left kidneys were taken for the determination of total kidney collagen, fluorescence, acid solubility of collagen and the right kidneys were used for the determination of glomerular basement membrane thickness. Total kidney collagen was significantly higher in the MG treated mice compared to control mice. Only about half the amount of collagen could be extracted from kidneys of MG treated animals indicating reduced solubility. Fluorescence in proteins from extracted kidneys of MG treated animals was about twice that of untreated animals. Glomerular basement membrane thickness was significantly higher in MG treated animals. Our findings indicate that MG can increase glomerular basement membrane thickness and the suggested underlying mechanism may be decreased solubility by increased cross linking as reflected by elevated protein fluorescence and decreased acid salt extraction. The involvement of MG in the development of diabetic complications postulated by others is herewith clearly supported by our findings. PMID- 9740318 TI - Endomorphin 1 and 2, the endogenous mu-opioid agonists, produce biphasic changes in systemic arterial pressure in the cat. AB - The endogenous peptides endomorphin 1 and 2 are newly isolated, potent, selective mu-opioid receptor agonists. In the present study, responses to the endomorphin peptides were investigated in the systemic vascular bed of the cat. Endomorphin 1 and 2 induced dose-related biphasic changes in systemic arterial pressure when injected in doses of 1-30 nmol/kg i.v. The biphasic responses to endomorphin 1 and 2 were characterized by an initial increase followed by a decrease in systemic arterial pressure. In terms of relative vasodepressor activity, endomorphin 1 and 2 were similar in potency and approximately 10-fold less potent than the ORL1 ligand nociceptin (orphanin FQ) in decreasing systemic arterial pressure. The biphasic arterial pressure changes in response to endomorphin 1 and 2 were inhibited by the opioid receptor antagonist naloxone in a dose of 2 mg/kg i.v. These results demonstrate that endomorphin 1 and 2 produce significant, naloxone-sensitive changes in systemic arterial pressure that are characterized by an initial increase followed by a secondary decrease in arterial pressure in the cat. PMID- 9740319 TI - Insulin-like 4 (INSL4) gene expression in human embryonic and trophoblastic tissues. AB - Polypeptide growth factors play an important role in the regulation of human embryonic development. Insulin-like 4 gene (INSL4) is a member of the insulin family, which includes insulin, IGF-I, IGF-II, relaxin, and INSL3. Using RT-PCR, we previously found abundant INSL4 mRNA in the human placenta. In this study, we examined the chronology and spatial expression of this gene in sections of human placenta and conceptus by means of in situ hybridization. Expression of the IGF II gene was studied as a positive control. INSL4 distribution was tissue- and cell-specific. Indeed, INSL4 mRNA was most abundant in syncytiotrophoblast cells. In fetal tissues, INSL4 mRNA was identified in the perichondrium of all four limbs, vertebrae, and ribs. Moreover, INSL4 mRNA was abundant in interbone ligaments. These findings indicate that the INSL4 gene may play an important role in trophoblast development and regulation of bone formation. IGF-II mRNA, in agreement with the literature, are mainly located in the mesodermal core in the villous trophoblast and in most embryonic tissues. PMID- 9740320 TI - Expression of cell death regulatory genes and limited apoptosis induction in avian blastodermal cells. AB - Apoptosis is a well-established cellular mechanism for selective cell deletion during development. However, little is known about the expression of an apoptotic pathway and its role in determining the relative sensitivity of the early, pre gastrula, avian embryo to stress-induced cell death. We examined the sensitivity of avian blastodermal cells to engage in apoptosis upon exposure to etoposide, a topoisomerase II-inhibitor that rapidly and efficiently induces apoptosis in many cell types. We found that while the blastodermal cells are capable of engaging in apoptosis, they are highly resistant to such induction with respect to both concentration of drug required and length of exposure, even when compared to a tumor cell line with a known multi-drug resistant phenotype. Additionally, we assessed the expression of several candidate regulatory genes in blastodiscs from infertile eggs (i.e., maternal RNA transcripts), blastodermal cells immediately following oviposition, and various stages of early development up to gastrulation. This analysis revealed that several genes whose products have anti apoptotic activity, including bcl-2, bcl-xL, hsp70, grp78 and the glutathione S transferases, are expressed as early as the stage 1 embryo in the newly oviposited egg. These transcripts are also found in the infertile blastodisc, suggesting a role for maternally derived transcripts in the protection of the oocyte and zygote. Significantly, constitutive levels of hsp70 mRNA exceeded those of the other anti-apoptotic genes in the blastodermal cells. These results contribute to an emerging picture of stress resistance at the earliest stages of avian embryo development which involves multiple anti-apoptotic genes that act at different regulatory points in the apoptotic cascade. PMID- 9740321 TI - Development of bovine embryos in single in vitro production (sIVP) systems. AB - Single in vitro production (sIVP) of embryos enables the study of developmental parameters of individual oocytes or embryos. Because several previously published sIVP systems showed varying levels of success, we attempted to design a simple, semidefined sIVP system that resulted in developmental rates similar to those obtained through group production (gIVP). In a 5 x 3 x 4 factorial experiment, 4200 oocytes were randomly assigned to combinations of various maturation (sIVM), fertilization (sIVF), and culture (sIVC) treatments based on media TCM199 (5 treatments), TALP (3 treatments), and SOF/aa/BSA (4 treatments), respectively. All sIVP steps were carried out in 10-12 microl drops under oil. Embryo development to blastocyst on days 7 and 8 of culture was determined and blastocyst cell numbers measured as an indicator of embryo quality. No interaction was found within any combination of sIVM, sIVF and sIVC treatments. Also, there was no difference in percentage of development to various stages for embryos in any of the sIVM or sIVF treatments (over all treatment combinations). However, when treatment combinations included charcoal-treated serum addition on day 5 of culture, a significant increase in development (39.0% total blastocysts/total oocytes vs. 22.7, 23.8 and 23.5% for the other 3 sIVC treatments, respectively; P < 0.001) and decrease in mean cell number (114.2 vs. 149.1, 150.5 and 143.7 cells, respectively; P < 0.001) was observed. These results are comparable to those routinely obtained in this laboratory with gIVP and establish standard conditions for individual embryo production. PMID- 9740322 TI - MAP kinase cascade, but not ERKs, activated during early cleavage of mouse embryos. AB - Mitosis in early embryos is independent of exogenous mitogens, although mitogen stimulations and subsequent activation of a mitogen-activated protein (MAP) kinase cascade are essential for the proliferation of somatic cells. The activation state of the MAP kinase cascade during early cleavage has never been reported. In the present study, factors involved in the MAP kinase cascade--Ras, Raf-1, 14-3-3, MEK, and ERKs--and their activation states were detected by immunoblotting during early cleavage of mouse embryos. We found the constant presence of these molecules in mouse early embryos and the activation of Raf-1 exclusively at the M-phase. An immunoprecipitation study revealed that active Raf 1 in the M-phase was dissociated from 14-3-3, as in somatic cells, whereas inactive Raf-1 was associated with 14-3-3. Surprisingly, the ERKs (MAP kinases) were not activated throughout early cleavage, although M-phase-specific activation of the MAP kinase kinase, MEK was observed. Myelin basic protein kinase activity was, however, significantly higher in the M-phase than in the interphase. These results indicate that the MAP kinase cascade is activated at the M-phase and that some MAP kinases other than ERKs are activated during early cleavage of mouse embryos. PMID- 9740323 TI - Role of sialic acid in the endocytosis of prosaposin by the nonciliated cells of the rat efferent ducts. AB - The present study examines the mechanism of endocytosis of testicular prosaposin by the nonciliated cells of the efferent ducts. Testicular prosaposin is secreted by Sertoli cells into the lumen of the seminiferous tubules as a 70 kDa isomer where it binds to the tail of spermatozoa. In the efferent ducts, after dissociating from the plasma membrane of the spermatozoa, prosaposin is endocytosed by the nonciliated cells, presumably by receptor-mediated endocytosis. The initial step of receptor-mediated endocytosis usually results from the binding of a ligand's terminal oligosaccharide to a receptor on the cell surface. Thus, in the present study, several monosaccharides were injected in the lumen of the efferent ducts to compete with the binding and endocytosis of prosaposin. A quantitative electron microscopic approach was utilized and the number of gold particles, indicating anti-prosaposin immunoreactive sites, were scored over the various cell compartments including the plasma membrane, endocytic vesicles, early endosomes, and late endosomes. The length of the plasma membrane and the areas of endocytic vesicles, early endosomes, and late endosomes were measured with an image analyzer and the number of grains expressed per microm (plasma membrane) and microm2 (endocytic vesicles/endosomes) respectively. The quantitative analysis was performed in untreated animals (controls) and animals treated with various sugars (i.e., glucose, galactose, mannose, mannose 6 phosphate, N-acetylglucosamine and N-acetylgalactosamine) injected into the lumen of the efferent ducts at a concentration of 20 mM. Sialic acid caused the greatest decrease in the labeling density of the endocytic elements. Mannose 6 phosphate also caused a decrease in labeling but to a lesser extent. Various amounts of sialic acid (0.02 mM, 0.2 mM, 2 mM, 20 mM, and 200 mM) showed that most of these concentrations produced a significant decrease in the labeling density of endocytic vesicles and endosomes. Moreover, Western blots of prosaposin isolated from seminiferous tubular fluids followed by glycan analysis with Sambucus nigra agglutinin (SNA) and Maackia amurensis agglutinin (MAA), revealed that this protein has sialic acid residues that are terminally linked to galactose and/or N-acetylgalactosamine (alpha-NeuNAc-[2->6]-Gal and alpha-NeuNAc [2->6]-GalNAc). These data indicate that testicular prosaposin is removed from the lumen of the efferent ducts by the noncialiated cells via a receptor that recognizes prosaposin's terminal sialic acid residues. PMID- 9740325 TI - Cloning and sequencing of cDNA encoding for a human sperm antigen involved in fertilization. AB - A cDNA encoding for a sperm antigen, designated NZ-2, was cloned and sequenced from human testis cDNA-lambda gt11 expression library by using antibodies to human sperm surface antigens belonging to 14-18 kD molecular regions. These sperm antigens are involved in binding to zona pellucida of the human oocyte. Computer generated translation analysis of 963-bp cDNA yielded an open reading frame (ORF) of 163 amino acids (aa) with first ATG, Met start codon at nucleotide (nt) 335 and the stop codon TAA at nt 824. The NZ-2 cDNA has 335-bp 5' and 139-bp 3' noncoding regions. The translated protein has a calculated molecular weight of approximately 19 kD, and has two casein kinase II (CK-2) sites at aa 94-97 and 149-152, respectively. Extensive computer search in the GenBank, National Biomedical Research Foundation (NBRF), and Swiss database indicates it to be a novel protein, having 99.5% nt sequence similarity, except for the first 40-bp, only with the human bacterial artificial chromosome (BAC) containing cloned human sperm DNA, at position 76935-76009. The in vitro translated product of T3 RNA polymerase by using NZ-2 cDNA digested with XhoI yielded a protein band of approximately 20 kD, indicating it to be sense strand. The in vitro translated product of T7 RNA polymerase by using NZ-2 cDNA digested with NotI did not yield any protein band, indicating it to be antisense strand. The approximately 20 kD protein was recognized specifically by the antisperm IgG, not by the control IgG in the Western blot procedure. Neither antisperm IgG nor control IgG recognized any protein band in the in vitro translation products of the antisense strand. The human genomic DNAs from three different cells/tissues namely, sperm, kidney, and testis when cut by HindIII, and then hybridized with the NZ-2 cDNA probe in the Southern blot procedure, showed restriction fragment length polymorphism (RFLP). The recombinant human sperm NZ-2 antigen may find applications in the development of a contraceptive vaccine, and diagnosis and treatment of infertility in humans. PMID- 9740324 TI - Mouse Odf2 cDNAs consist of evolutionary conserved as well as highly variable sequences and encode outer dense fiber proteins of the sperm tail. AB - The outer dense fibers (ODF) of the mammalian sperm tail comprise a unique, specialized, and very prominent structure, consisting of nine fibers surrounding the axoneme. The ODF may play an important but as yet undefined role in sperm morphology, integrity and function. Study of the ODF is hampered by insufficient knowledge of their protein composition and the genetic regulation of their synthesis. We report here on the characterization of cDNAs encoding the Odf2 proteins of outer dense fibers of mouse sperm. We isolated two cDNA clones with variable 5' regions. Variability in sequence is restricted to specific regions in the N-terminal part of the encoded proteins, whereas the C-terminal part is highly conserved in Odf2 proteins both between species and within a species. This variability is confirmed at the protein level. The outer dense fibers could be detected immunologically in total sperm tails allowing a direct comparison of their length in relation to the length of the sperm tail. Odf2 transcripts could be demonstrated in testicular RNA and are restricted to germ cells. The start of transcription is in step 5 spermatids of tubular stage V and the RNA could be detected in the cytoplasm of differentiating spermatids in all subsequent tubular stages. PMID- 9740326 TI - Site-directed mutagenesis of boar proacrosin reveals residues involved in binding of zona pellucida glycoproteins. AB - Proacrosin, the zymogen form of the serine protease beta-acrosin, is thought to function as a secondary binding molecule between mammalian gametes during fertilization (Jansen et al., 1995: Int J Dev Biol 39, 501-510). The interaction involves strong ionic bonds between positively charged amino acids on proacrosin and negatively charged polysulphate groups on zona pellucida glycoproteins. In this investigation, we identified the basic residues on proacrosin that are important for this binding. Site-directed mutagenesis shows that two groups of amino acids comprising His47, Arg50, and Arg51 together with Arg250, Lys252, and Arg253 are crucial because their deletion or replacement severely reduces affinity for zona glycoproteins. Molecular models of proacrosin reveal that these residues are located along one face of the protein on two exposed surface loops that project over and around the catalytic site. These findings support the hypothesis that polysulphate binding sites on proacrosin are formed by a restricted number of basic amino acids on the surface of the protein, presenting a specific orientation that is complementary to negatively charged sulphate groups on zona glycoproteins. Identification and elucidation of the stereochemistry of these charged moieties will aid design of new kinds of nonsteroidal antifertility agents. PMID- 9740327 TI - Interactions between a decapacitation factor and mouse spermatozoa appear to involve fucose residues and a GPI-anchored receptor. AB - Epididymal mouse spermatozoa have a surface-associated decapacitation factor (DF) that can be removed precociously by centrifugation, resulting in acceleration of capacitation and increased fertilizing ability. Addition of exogenous DF to capacitated suspensions inhibits fertilizing ability and reverses capacitation in acrosome-intact cells. DF appears to regulate a Ca2+-ATPase, located primarily in the post-acrosomal region. The present investigations of DF<-->spermatozoon interaction indicate that DF can be removed from uncapacitated cells by treatment with phosphatidylinositol-specific phospholipase C (PIC), suggesting the involvement of a glycosylphosphatidylinositol (GPI) moiety. However, exogenous DF cannot reassociate with PIC-treated spermatozoa, suggesting that DF may bind to spermatozoa via a GPI-anchored receptor. DF binding appears to involve fucose residues, since depletion of endogenous DF followed by brief exposure to fucose (0.1-10 mM) prevented DF reassociation with cells. Furthermore, 5 mM fucose could displace DF from uncapacitated cells, accelerating capacitation and resulting in a higher proportion of fertilized oocytes, with increased polyspermy, than obtained with untreated controls. FITC-labelled fucosylated BSA bound specifically to the postacrosomal region, binding being inhibited by both excess fucose and crude DF. UEA I, a lectin with specificity for fucose residues, bound to the postacrosomal region of cells preincubated in fucose but not crude DF, and blocked DF binding to DF-depleted cells. These results are consistent with the DF binding, via fucose residues, to a GPI-anchored receptor. Fucose binding sites are in the same region where Ca2+-ATPase, the enzyme regulated by DF, has been localized; these results support the hypothesis that DF modulates capacitation by regulating enzyme activity and hence the intracellular Ca2+ concentration. PMID- 9740328 TI - Quantitative changes in sperm head morphology during passage through the male excurrent duct system of the rabbit. AB - A fine adjustment of sperm head size and shape occurs during maturation and storage within the male excurrent duct of the rabbit. This remodelling, as judged by morphometric values of area, perimeter, length, width, and shape factors, takes place mostly in passage from the seminiferous tubules of the testis to the distal caput of the epididymis. The dimensions of sperm heads from the distal corpus of the epididymis break the general tendency toward a reduction in size and more elliptical shapes. A period of transport and storage within the epididymal cauda and vas deferens follows in which there are no further changes in sperm head morphometry. It can be concluded that the period immediately following sperm release from the testis is crucial to the final morphological maturation of spermatozoa. Moreover, the fact that changes are detected in the appearance of sperm heads at successive stages of sperm maturation suggests that the dimensions of a particular epididymal spermatozoon may be taken as an approximate indication of its relative maturity. PMID- 9740329 TI - c-Abl proto-oncoprotein is expressed and tyrosine phosphorylated in human sperm cell. AB - The presence and possible role of c-Abl proto-oncoprotein was investigated in human sperm cell. The c-Abl monoclonal antibody (mAb), against the protein tyrosine kinase domain of v-Abl protein, reacted specifically with the acrosomal region of methanol-fixed capacitated and non-capacitated human sperm cell in the indirect immunofluorescence technique. The c-Abl mAb predominantly recognized two protein bands of 145 kD and 95 kD in detergent-solubilized (Triton X-100 and NP 40) sperm and testes preparations in the Western blot procedure. The 95 kD protein band reacted stronger than the 145 kD band and was the only band detected in the lithium diiodosalicylate (LIS)-solubilized sperm preparation, and even in the Triton X-100/NP-40 extracts of sperm of some men. In the in vitro kinase assay using the Triton X-100-solubilized capacitated sperm preparation, the 95 kD protein was autophosphorylated at the tyrosine residues, which was inhibited in the presence of c-Abl mAb. The tyrosine phosphorylation of sperm proteins, especially of the 95 kD protein, has been shown to have a vital role in human sperm function, namely, the sperm capacitation/acrosomal exocytosis and binding to zona pellucida of oocyte. These findings suggest that the c-Abl or c-Abl-like proteins are present in mature sperm cells that are tyrosine autophosphorylated and may have a role in human sperm cell function. PMID- 9740331 TI - Intercellular adhesion molecule (ICAM-1) and the risks of developing atherosclerotic disease. PMID- 9740330 TI - Effects of differentiation on the transcriptional regulation of the FGF-4 gene: critical roles played by a distal enhancer. AB - Embryonal carcinoma (EC) cells are used widely as a model system for studying the expression of developmentally regulated genes, in particular genes that are regulated at the transcriptional level when EC cells differentiate. This review focuses on the molecular mechanisms that govern the transcription of the fibroblast growth factor-4 (FGF-4) gene, which appears to be the first FGF expressed during mammalian development. Interest in this gene has increased considerably with the finding that FGF-4 is essential for mammalian embryogenesis. The FGF-4 gene has also generated considerable interest because it is inhibited at the transcriptional level when EC cells undergo differentiation and because this gene is regulated by a powerful distal enhancer located 3 kb downstream of the transcription start site in the last exon of the gene. Hence, study of the FGF-4 gene is likely to shed light on the molecular mechanisms by which distal enhancers regulate gene expression. In addition to being regulated by the downstream enhancer, the expression of this gene is influenced by a regulatory region located just upstream of the transcription start site, which contains two Sp1 motifs and a CCAAT box motif. Examination of the downstream enhancer has identified three functional cis-regulatory elements: a high mobility group (HMG) protein binding motif, an octamer binding motif, and an Sp1 motif, which are likely to bind Sox-2, Oct-3, and Sp1/Sp3, respectively, in vivo. Interestingly, Sox-2 and Oct-3 expression, like FGF-4 expression, decreases when EC cells differentiate, which suggests that the loss of these transcription factors is responsible, at least in part, for the transcriptional turn-off of the FGF-4 gene. In view of these and other findings, we present a model for the differential expression of the FGF-4 gene that includes not only the contributions of specific transcription factors, but also the contribution of chromatin structure before and after differentiation. PMID- 9740332 TI - Is there MUSIC in IVUS guided stenting? Is this MUSIC going to be a MUST? Multicenter Ultrasound Stenting in Coronaries Study. PMID- 9740333 TI - Improving outcomes in heart failure. PMID- 9740334 TI - Diastolic dysfunction in hypertrophic cardiomyopathy. PMID- 9740335 TI - Intravascular ultrasound-guided elective stent implantation in calcified coronary lesions. A picture is worth more than a thousand words (sometimes!) PMID- 9740336 TI - Heparin and platelet reactivity in coronary artery disease. PMID- 9740337 TI - Applying the open artery theory: use of predictive survival markers. PMID- 9740338 TI - The pre-hospital management of acute heart attacks. Recommendations of a Task Force of the The European Society of Cardiology and The European Resuscitation Council. PMID- 9740339 TI - Driving and heart disease. PMID- 9740340 TI - The search for novel antiarrhythmic strategies. Sicilian Gambit. PMID- 9740341 TI - Classical risk factors and their impact on incident non-fatal and fatal myocardial infarction and all-cause mortality in southern Germany. Results from the MONICA Augsburg cohort study 1984-1992. Monitoring Trends and Determinants in Cardiovascular Diseases. AB - BACKGROUND: The MONICA (Monitoring Trends and Determinants in Cardiovascular Diseases) project in Augsburg provides the first population-based cohort study in Germany to quantify the associations of the risk factors hypertension, hypercholesterolaemia and smoking with incident non-fatal and fatal myocardial infarction and all-cause mortality, and to assess their impact at the population level. METHODS: The cohort comprises 1074 men and 1013 women aged 45-64 years; they were followed over 8 years from 1984-1992. In the men, there were 61 non fatal and fatal myocardial infarctions and 92 all-cause mortality events over this period; in the women the number of deaths from all causes was 45. Incidence rates, hazard rate ratios, population attributable fractions and rate advancement periods were calculated. RESULTS: Adjusting for confounders, the myocardial infarction hazard rate ratios for men with hypertension, or a total cholesterol/HDL-cholesterol ratio > or =5.5, or smoking > or =20 cigarettes/day, were 2.0 (95% CI 1.2-3.5), 2.9 (95%, CI 1.7-5.0), and 2.7 (95% confidence interval (CI) 1 4-5.0), respectively. The risk factor combination total cholesterol/HDL cholesterol ratio > or = 5.5 and cigarette smoking was particularly hazardous. The three risk factors contributed 65% of the burden of myocardial infarction in the population. The rate advancement period for myocardial infarction associated with hypertension, total cholesterol/HDL cholesterol ratio > or =5.5 or smoking > or =20 cigarettes/day was 8.3, 12.4 and 11.5 years, respectively. In women, these risk factors were similarly predictive of all-cause mortality. Comparing the cohort data from Augsburg with those of two occupational cohorts from Germany reveals higher absolute myocardial infarction risks in the Augsburg population; however, the relative risk estimates in the Augsburg and the two occupational cohorts were very similar. CONCLUSION: Our results confirm the important contribution of the classical risk factors to the risk of myocardial infarction and all-cause mortality in Germany. The results pertaining to the concept of rate advancement periods particularly demonstrate the great potential for prevention. PMID- 9740342 TI - Long-term follow-up after early intervention with intravenous diltiazem or intravenous nitroglycerin for unstable angina pectoris. AB - AIMS: In a double-blind randomized trial in unstable angina it was shown that intravenous diltiazem reduced ischaemic events in the first 48 h after inclusion better than intravenous nitroglycerin. The present study was performed to establish the long-term prognosis of the randomized patients, with respect to their initial treatment assignment. METHODS AND RESULTS: One year follow-up data on ischaemic end-points and anti-ischaemic medication were recorded. Results were available for all of the 121 randomized patients. One hundred and sixty-seven primary endpoint events were recorded, of which 54 occurred in the first 48 h and 113 during the follow-up. Survival analysis showed that event-free survival was significantly better in the diltiazem group (45.0%) than in the nitroglycerin group (34.4%), P=0.04. The incidence rate after 48 h and one year for cardiac death are, respectively, 0% and 4.1%. The trend in anti-ischaemic medication was higher in the nitroglycerin group. For beta-blockers, this trend became significant after 12 months (P=0.03). CONCLUSION: These results show that the initial benefit obtained by early treatment with intravenous diltiazem was preserved during the first year after the initial hospitalization, and that, despite the high risk of cardiac events in our population, the overall mortality 12 months after inclusion was low. PMID- 9740343 TI - Intravascular ultrasound-guided optimized stent deployment. Immediate and 6 months clinical and angiographic results from the Multicenter Ultrasound Stenting in Coronaries Study (MUSIC Study) AB - OBJECTIVES: A study was set up to validate the safety and feasibility of intravascular ultrasound-guided stenting without subsequent anticoagulation, and its impact on the 6 months restenosis rate. METHODS: The study was designed to be multicentred, prospective, and observational. RESULTS: One hundred and sixty-one patients with stable angina and a de novo coronary artery lesion were enrolled. In four patients, the implantation of a Palmaz-Schatz (with spiral bridge) stent had failed. One of these four patients died 3 days following bypass surgery. In two other patients, intravascular ultrasound assessment was not performed. One hundred and twenty-five of the remaining 155 patients (81%) were treated with aspirin (100 mg x day(-1)), because all three criteria for optimized stent expansion were met. Twenty-two of the remaining 38 patients (25%), in whom at least one criterion was not met were treated with aspirin and acenocoumarol (3 months, INR 2.5-3.5), while 16 patients only received aspirin. Stent thrombosis was documented in two patients (1.3%) for which repeat angioplasty was performed. During the hospital stay, there were no deaths or Q-wave myocardial infarctions. Five patients (3.2%) sustained a non-Q-wave myocardial infarction. During the follow-up period (198+/-38 days, complete for all patients, except one), one patient (0.6%) sustained a Q-wave myocardial infarction, one (0.6%) underwent bypass surgery, and repeat angioplasty was performed in nine patients (5.7%). In two of the nine patients, repeat angioplasty involved another lesion. Therefore, the target lesion revascularization rate during follow-up was 4.5% (seven patients). At quantitative coronary angiography, the minimal lumen diameter (mean+/-SD) increased from 1.12+/-0.34 mm before to 2.89+/-0.35 mm after stenting. Repeat angiography at 6 months was performed in 144 patients (92%). The minimal lumen diameter at follow-up was 2.12+/-0.67 mm. Restenosis (diameter stenosis of 50% or more) was documented in 12 patients or 8.3%. When the two patients with documented stent thrombosis are included, the restenosis rate amounts to 97%. CONCLUSIONS: These data confirm that, in selected patients, stents can safely be implanted without the use of systemic anticoagulation, provided optimal stent expansion is achieved. The exact role of intravascular ultrasound in the achievement of these results needs to be established by appropriately designed studies. In the meantime, intravascular ultrasound coupled with the Palmaz-Schatz stent incorporating a spiral bridge, may have contributed considerably to the immediate angiographic outcome, which in turn may explain the favourable clinical and angiographic outcome at 6 months. PMID- 9740344 TI - Treatment of calcified coronary lesions with Palmaz-Schatz stents. An intravascular ultrasound study. AB - AIMS: To evaluate the result of coronary stenting in calcified lesions and to find morphological and procedural factors influencing the final result. METHODS AND RESULTS: Three hundred and twenty three native coronary artery lesions in 303 patients (197 men, mean age 63.9 +/- 11.5 years) treated with Palmaz-Schatz stents were differentiated into four groups depending on their degree of circumferential calcification as defined by intravascular ultrasound [0-90 degrees (n=120), 91-180 degrees (n=58, 181-270$ (n=71) and 271-360 degrees n=74)]. In 117 lesions rotational atherectomy was used prior to stent placement. Intravascular ultrasound and quantitative angiography were performed prior to treatment and after stent placement to measure minimal and maximal lumen diameter and lumen cross-sectional area at the lesion site and the reference segments. Acute lumen gain and eccentricity index were calculated. Although higher balloon pressures were used than in the minimally calcified lesions. the final angiographic minimal lumen diameter decreased with increasing arc of calcification (3.01 +/- 0.47, 3.04 +/- 0.43, 2.85 +/- 0.53, 2.83 +/- 0.40 mm, respectively, P=0.0320) resulting in a decrease in acute diameter gain with increasing arc of calcification (2.06 +/- 0.51, 1.91 +/- 0.46, 1.81 +/- 0.56, 1.78 +/- 0.51 mm, respectively, P=0.0067). Adjunctive rotational atherectomy prior to stent placement resulted in a greater acute diameter and a greater lumen cross-sectional area gain, coupled with less final residual stenosis than pre treatment with balloon angioplasty. CONCLUSION: Implantation of stents in calcified lesions results in less optimal stent expansion, especially in lesions with thick, eccentric calcific plaque layers. Use of adjunctive rotational atherectomy before stent placement may improve the procedural result. PMID- 9740345 TI - Incidence, outcome and prediction of early clinical events following percutaneous transluminal coronary angioplasty. A comparison between treatment with reviparin and unfractionated heparin/placebo (results of a substudy of the REDUCE trial). AB - BACKGROUND: Unfractionated heparin and its low molecular weight fragments possess antithrombotic properties, properties that are routinely exploited in coronary angioplasty (PTCA). OBJECTIVES: In the setting of the REDUCE trial, a randomized, double-blind, multicentre trial, the occurrence of acute or early clinical events was compared in patients treated with either unfractionated heparin/placebo or low molecular weight heparin (reviparin). METHODS AND RESULTS: Six hundred and twelve patients with native coronary artery obstructions randomized between unfractionated heparin/placebo and reviparin, were analysed. Baseline characteristics were similar in both groups. Using the intention-to-treat analysis, major acute or early events (myocardial infarction, re-PTCA, bypass surgery, death) occurred in 42 patients (7%), 29 in the control group and 13 in the treatment group (P=0.027). In order to develop a predictive model for the risk of early events following coronary balloon angioplasty, clinical as well as pre-PTCA and procedural characteristics were analysed. Thrombi at the treated lesion site (P=0.02), dissection (P<0.001), lesion type B2 and C according to the NHLBI classification (P<0.001), diameter stenosis >50% post-PTCA (P<0.001), and length of stenosis >20mm (P=0.005) were significantly associated with the occurrence of acute events. By multiple logistic regression analysis, in which these variables and the treatment regimen were entered, dissection (P=0.042), diameter stenosis >50% (P<0.028) and lesion type B2 and C (P=0.017) were found to be independently predictive of early adverse events. Bleeding complications were similar in the two treatment groups. CONCLUSIONS: Reviparin, given in a very early stage of vascular injury, compares favourably with unfractionated heparin/placebo, by reducing abrupt closure and acute-phase adverse outcome following PTCA. With respect to the evaluated risk factors for acute events, the positive effect of reviparin on early adverse outcome after PTCA may be due to improved antithrombotic properties as compared to unfractionated heparin. PMID- 9740346 TI - Increased platelet responsiveness following coronary stenting. Heparin as a possible aetiological factor in stent thrombosis. AB - AIMS: Platelet activation may be a determinant of thrombotic and restenotic complications following intracoronary stenting. In order to measure the effect of stenting on platelet activation antigen expression we used whole blood flow cytometry in 18 patients undergoing Palmaz-Schatz stenting (treated with full anticoagulation) and compared these with a group of 18 patients undergoing elective angioplasty. The effects of low molecular weight heparin and unfractionated heparin on platelet behaviour were also studied, both in vitro and in vivo to determine the contribution of prolonged heparin therapy to platelet activation following stenting. METHODS AND RESULTS: Fibrinogen binding to activated GPIIb-IIIa, and surface expression of P-selectin, GPIb and GPIIb-IIIa antigens were measured in unstimulated peripheral blood samples (rest) and on stimulation with adenosine diphosphate (0.1-10 micromol x 1(-1)) and thrombin (0.02-0.16 U x ml(-1)). No changes were seen in resting samples following angioplasty or stenting. Agonist responsiveness was unaltered after angioplasty, but in stented patients antigen expression in response to thrombin was significantly reduced (P< or =0.04), whilst the adenosine diphosphate response was significantly increased (P=0.01). Similar effects were observed in patients with unstable angina treated with either low molecular weight heparin or unfractionated heparin in vivo. In vitro, both unfractionated and low molecular weight heparin inhibited thrombin-induced platelet activation, but stimulation of adenosine diphosphate responses was more marked with unfractionated than low molecular weight heparin. CONCLUSIONS: There was a significant increase in platelet responsiveness to adenosine diphosphate following intracoronary stenting in patients treated with conventional anticoagulants. This was probably a consequence of treatment with heparin. Activation of platelets by heparin may explain the increased rate of stent thrombosis in patients treated with anticoagulant therapy. Low molecular weight heparins stimulate platelets less than unfractionated heparin. PMID- 9740347 TI - A randomized controlled trial of inspiratory muscle training in stable chronic heart failure. AB - AIMS: To assess whether a domiciliary programme of specific inspiratory muscle training in stable chronic heart failure results in improvements in exercise tolerance or quality of life. METHODS AND RESULTS: We conducted a randomized controlled trial of 8 weeks of inspiratory muscle training in 18 patients with stable chronic heart failure, using the Threshold trainer. Patients were randomized either to a training group inspiring for 30 min daily at 30% of maximum inspiratory mouth pressure, or to a control group of 'sham' training at 15% of maximum inspiratory mouth pressure. Sixteen of the 18 patients completed the study. Maximum inspiratory mouth pressure improved significantly in the training group compared with controls, by a mean (SD) of 25.4 (11.2) cmH2O (P=0.04). There were, however, no significant improvements in treadmill exercise time, corridor walk test time or quality of life scores in the trained group compared with controls. CONCLUSION: Despite achieving a significant increase in inspiratory muscle strength, this trial of simple domiciliary inspiratory muscle training using threshold loading at 30% of maximum inspiratory mouth pressure did not result in significant improvements in exercise tolerance or quality of life in patients with chronic heart failure. PMID- 9740349 TI - Pattern of left ventricular filling in hypertrophic cardiomyopathy. Assessment by Doppler echocardiography and radionuclide angiography. AB - AIMS: The left ventricle in hypertrophic cardiomyopathy is anatomically and functionally non-uniform. This study was undertaken to verify whether a heterogeneity in the pattern of diastolic filling can be detected along the left ventricular inflow tract in hypertrophic cardiomyopathy. METHODS AND RESULTS: Early (E) and late (A) diastolic velocities were recorded by Doppler echocardiography at mitral and at mid-ventricular level in 16 normal volunteers and 30 patients with hypertrophic cardiomyopathy. Patients with hypertrophic cardiomyopathy also underwent radionuclide angiography to assess left ventricular function. E wave decreased significantly in normal volunteers (80 +/- 15 to 60 +/ 14 cm x s(-1); P<0.001), but it increased in hypertrophic cardiomyopathy (76 +/- 22 to 87 +/- 28 cm x s(-1) P=0.04), whereas the A wave decreased similarly in both. By multivariate analysis, systolic asynchrony and the ejection fraction of left ventricular lateral wall were directly related to the pattern of early filling progression (r=0.656, F=9.467; P<0.002). Moreover, systolic asynchrony showed a univariate direct correlation with changes in E velocity (r=0.42; P=0.02). CONCLUSION: Many patients with hypertrophic cardiomyopathy have an acceleration of filling within the left ventricular inflow tract; this phenomenon is directly related to systolic asynchrony and ejection fraction of the left ventricular lateral wall, suggesting increased suction. PMID- 9740348 TI - Feasibility of a nurse-monitored, outpatient-care programme for elderly patients with moderate-to-severe, chronic heart failure. AB - AIMS: To evaluate the feasibility of a nurse-monitored, outpatient-care program for elderly patients previously hospitalized with chronic heart failure. METHODS AND RESULTS: Patients with chronic heart failure hospitalized in the medical wards were screened to find those eligible for a randomized study to compare the effect of a nurse-monitored, outpatient-care programme aiming at symptom management, with conventional care. The inclusion criteria were patients classified in New York Heart Association classes III-IV, age 65 years, and eligibility for an outpatient follow-up programme. The total in-hospital population of patients discharged with a heart-failure diagnosis was surveyed. Eighty-nine per cent of all the hospitalized patients (n=1541) were 65 years old. Of these, 69% (n=1058) were treated in the medical wards which were screened. The study criteria were met by 158 patients (15%). No visits to the nurse occurred in 23 cases among the 79 patients randomized to the structured-care group (29%), mainly on account of death or fatigue. The numbers of hospitalizations and hospital days did not differ between the structured-care and the usual-care groups. CONCLUSIONS: Given the selection criteria and the outline of the interventions, the outpatient, nurse-monitored, symptom-management programme was not feasible for the majority of these elderly patients with moderate-to-severe, chronic heart failure, mainly because of the small proportion of eligible patients and the high drop-out rate. Management of these patients would have to be more adjusted to their home situation. PMID- 9740350 TI - Modification of cytokine patterns in subjects bearing the HLA-B8,DR3 phenotype: implications for autoimmunity. AB - The factors influencing the pathogenesis of autoimmune disease are not fully known, but the host genotype undoubtedly plays a role in determining the outcome of these diseases. The role of the host's major histocompatibility complex (MHC) genotype in the regulation of susceptibility to autoimmune diseases has been extensively studied in different populations, and certain HLA (the human MHC) alleles and haplotypes have been reported to be associated with several autoimmune diseases. In particular, the association with genes from the HLA B8,DR3 haplotype has been reported by different research groups. This haplotype is associated in all Caucasian populations with a wide variety of diseases with autoimmune features, and in healthy subjects it is associated with a number of immune system dysfunctions. Mainly, peripheral blood mononuclear cells from HLA B8,DR3-positive and -negative individuals differ in their ability to produce interleukin (IL)-2, IL-5, IL-12 and interferon-gamma upon stimulation with the mitogen phytohaemoagglutinin (PHA), while producing similar amounts of IL4, IL-6 and IL-10. Furthermore, in HLA-B8,DR3-positive subjects tumor necrosis factor alpha secretion is increased both with and without PHA stimulation. Accurate control of the functional repertoire of an immune response is a critical parameter in the response to infections as well as in immunopathology. MHC control of the class of the immune response at the level of cytokine production is a sophisticated way in which this occurs. This control might be involved in adaptive immune responses to infections as well as in immunopathology. PMID- 9740352 TI - Cytokines and the mechanisms of action of vaccine adjuvants. AB - There is an increasing trend away from classical attenuated or killed whole pathogen vaccines towards developing chemically defined preparations. Paradoxically, in order to be effective, these defined vaccines require incorporation into adjuvants, of which very little is actually understood about how or why they work. It is, therefore, of little surprise that, in this context, adjuvants have been referred to as 'the immunologists' dirty little secret'. [Janeway CA, Approaching the asymptote? Evolution and revolution in immunology. Cold Spring Harbor Symp Quantit Biol (1989) 54: 1-13]. However, modern techniques in immunology have made available a number of powerful tools that will allow a more complete dissection of how traditional adjuvants work, knowledge of which will not only facilitate the rational design of new adjuvants but also further clarify our understanding of the immune system. The initial studies described in this review indicate that cytokines play an important, if not the central, role in the ability of adjuvants to influence both the quantity and quality of immune responses. PMID- 9740351 TI - Early expression of interleukin-12, p40 subunit and IFN-gamma inhibits regression of AK-5 tumor. AB - Differential immune response of syngeneic animals to a rat histiocytoma AK-5 based on the route of transplantation was investigated. Spontaneous regression of subcutaneous tumor was observed in 55-60% of animals. On the other hand, when the tumor cells were injected intraperitoneally, none of the animals survived. Earlier studies from this laboratory indicated upregulation of Th-1-type cytokines, leading to early tumor regression when the tumor was transplanted subcutaneously. Hence we evaluated and compared the circulatory-cytokine profiles in both s.c. and i.p. tumor-injected animals. Our results show an early increase in the p40 subunit of IL-12, prolific increase in IFN-gamma and lower levels of IL-2 in i.p. tumor-injected animals. However, there were no significant differences in the levels of transcripts for these cytokines in either of the groups. Significantly, a lower level of cytotoxicity was observed with splenocytes from i.p. tumor-transplanted animals. Moreover, the cytotoxicity of IL-12-activated but not IL-2-activated NK cells was inhibited by sera (rich in IL 12, p40 subunit) from i.p. tumor-transplanted animals, suggesting the participation of p40 subunit in the regulation of tumor regression. Thus the present study suggests a possible translational regulation of Th-1-type cytokines in AK-5 tumor-host interaction. PMID- 9740353 TI - Cytokines and therapeutic oligonucleotides. AB - Therapeutic oligonucleotides - short strands of synthetic nucleic acids - encompass antisense and aptamer oligonucleotides. Antisense oligonucleotides are designed to bind to target RNA by complementary base pairing and to inhibit translation of the target protein. Antisense oligonucleotides enable specific inhibition of cytokine synthesis. In contrast, aptamer oligonucleotides are able to bind directly to specific proteins. This binding depends on the sequence of the oligonucleotide. Aptamer oligonucleotides with CpG motifs can exert strong immunostimulatory effects. Both kinds of therapeutic oligonucleotides - antisense and aptamer oligonucleotides - provide promising tools to modulate immunological functions. Recently, therapeutic oligonucleotides have moved towards clinical application. An antisense oligonucleotide directed against the proinflammatory intercellular adhesion molecule 1 (ICAM-1) is currently being tested in clinical trials for therapy of inflammatory disease. Immunostimulatory aptamer oligonucleotides are in preclinical development for immunotherapy. In the present review we summarize the application of therapeutic oligonucleotides to modulate immunological functions. We include technological aspects as well as current therapeutic concepts and clinical studies. PMID- 9740354 TI - The role of cytokines in acute graft-versus-host disease. AB - Graft-versus-host disease (GVHD) remains the principal complication limiting the wider application of allogeneic bone marrow transplantation (BMT). Advances in basic immunology during the last decade have demonstrated how interactions between immunologically competent cells are governed by cytokines, and much recent research has focused on the roles of these mediators in the pathogenesis of acute GVHD. This article reviews current evidence that dysregulated cytokine production can be considered a cascade of sequential monocyte and T-cell activation that is responsible for many of the manifestations of acute GVHD. We suggest that cytokine dysregulation can be conceptualized in three phases. Phase 1 is initiated by the conditioning of the host, which induces inflammatory processes in recipient tissues. Donor T-cell activation by host alloantigens and subsequent cytokine secretion in phase 2 is facilitated by the consequences of phase 1. The T-cell-derived cytokines of phase 2 activate distal inflammatory mediators, which, in synergy with T- and NK-cell-mediated cytotoxicity, produce the systemic morbidity of GVHD-associated immunosuppression in phase 3. Data from both experimental and clinical studies involving cytokines and their blockade in the prevention or treatment of GVHD are reviewed. PMID- 9740355 TI - Inflammatory bowel disease: potential therapeutic strategies. AB - This review deals with potential and possibly primary therapeutics that, through insight into the inflammatory cascade, result in more rational treatment principles replacing the classical therapy of inflammatory bowel disease (IBD), i.e. Crohn's disease (CD) and ulcerative colitis (UC). These new therapies might be useful for IBD patients, especially since the 'classical therapy' with agents like glucocorticoids, sulfasalazine, mesalazine, azathioprine, 6-mercaptopurine, cyclosporin and methotrexate is often only moderately effective and may have important side-effects. Controlled trials of the novel agents mentioned in this review have not yet been performed, however. PMID- 9740356 TI - Sentence context and lexical ambiguity resolution by the two hemispheres. AB - A lexical decision experiment investigated hemisphere asymmetries in resolving lexical ambiguity within a sentence context. Sentences that biased a single meaning (either dominant or subordinate) of sentence-final ambiguous words were followed by a lateralized target related to the sentence-congruent or incongruent meaning of the ambiguous word, or an unrelated word. In the RVF sentence-congruent targets were facilitated, while incongruent targets were not primed. In contrast, related targets were facilitated in the LVF, regardless of sentence context. This suggests that selecting the contextually appropriate word meaning requires the left hemisphere, and supports a right hemisphere role in maintaining alternate word senses. PMID- 9740357 TI - Selective spatial memory impairment after right unilateral temporal lobectomy. AB - Deficits in performance of both spatial and visual tasks are common following tissue loss in the right temporal lobe. Since spatial and visual attributes are frequently confounded in experimental tasks, we have studied patients following unilateral temporal lobectomy, in an attempt to determine which aspect mediates the observed deficits. Spatial and visual memory performance was compared in normal controls (n = 16), left temporal (LTL; n = 19) and right temporal (RTL; n = 19) lobectomy patients, by presentation of eight abstract designs in a spatial array for subsequent recall and recognition of the designs (visual memory) and recall of their spatial position (spatial memory). By varying the retention intervals for each group, all three groups were matched on both recall and recognition of the designs at sub-ceiling levels. In contrast, recall of the position of the designs (spatial memory), tested at equivalent delays to those of the visual memory tests, revealed a deficit in the RTL patients compared to both controls and LTL patients (p < 0.05). Magnetic resonance imaging (MRI) was used to quantify the extent of resection of the hippocampus and parahippocampal regions in the two patient groups and showed a significant correlation between hippocampal and parahippocampal removal and spatial memory in the RTL group only. These data support the notion of a disproportionately large involvement of the right hippocampus and adjacent regions in spatial memory. PMID- 9740358 TI - Normal intrasylvian anatomical asymmetry in children with developmental language disorder. AB - Symmetry of posterior intrasylvian cortices (e.g., planum temporale, planum parietale) has been suggested to represent a risk factor for developmental disorders of language and reading. Using high-resolution magnetic resonance morphometry, we studied 21 right-handed children with developmental language disorder of the phonologic-syntactic type, and found normal left-right asymmetry of the planum temporale and planum parietale when compared with 21 matched controls. The planum temporale was bilaterally smaller in the affected children, a finding accounted for by their approximately 7% smaller forebrain size. Our data do not support a role of gross visible unilateral or bilateral abnormalities of posterior intrasylvian ontogenesis in this disorder. PMID- 9740359 TI - Hemispheric dominance in the processing of J.S. Bach fugues: a transcranial Doppler sonography (TCD) study with musicians. AB - Although the majority of studies on musical processing in musicians observed a left hemisphere dominance which has usually been explained by a proficient analytical strategy used by these subjects, the findings are still inconsistent. Changes in hemispheric activity induced by listening to music (J. S. Bach fugues) and by recognizing the repetitions of the fugue theme were examined, using the technique of bilateral transcranial Doppler sonography (TCD) of the left and right middle cerebral artery (MCA). Subjects were 32 right-handed musicians, half of whom were members of an orchestra or members of a choir. The fugues were presented in two versions: a-cappella and instrumental. During passive listening to the a-cappella version, a weak left-dominant asymmetry of blood flow acceleration was observed, while there was no hemispheric asymmetry during listening to the instrumental version. During the task of fugue theme recognition, a highly significant asymmetry in favour of the right MCA was observed with both versions. It is concluded that when the processing of complex musical material has to be based on the analysis of melodic contour features and calls for working memory capacities a right hemisphere dominance is observed even in musically sophisticated subjects. PMID- 9740360 TI - Verbal and music dichotic listening tasks reveal variations in functional cerebral asymmetry across the menstrual cycle that are phase and task dependent. AB - Two dichotic listening tests, one a verbal consonant-vowel identification task, the other a musical chord recognition task, were administered to 32 women at two points during the menstrual cycle, menses (when oestrogen is low) and the midluteal phase (when oestrogen is high), in a counterbalanced repeated measures design. The degree of asymmetry changed across the cycle for both syllables and music. The right ear advantage recorded for the verbal task was greater during the midluteal phase than during menses. The left ear advantage recorded for the music task was greater during menses than during the midluteal phase. These reciprocal changes in asymmetry were the result of consistent changes in ear performance. From menses to the midluteal phase, left ear (right hemisphere) performance fell significantly for both tasks whereas right ear (left hemisphere) performance showed a small, but non-significant, increase. The findings are discussed in the light of evidence for phasic activational effects of gonadal steroids on both asymmetry and cognition which provide an explanation for the sometimes elusive nature and small effect size of sex differences in these characteristics. The relationships between sex differences in asymmetry and cognition are re-examined. PMID- 9740361 TI - Intensity coding of auditory stimuli: an fMRI study. AB - The effect of stimulus intensity (sound pressure level, SPL) of auditory stimuli on the BOLD response in the auditory cortex was investigated in 14 young and healthy subjects, with no hearing abnormalities, using echo-planar, functional magnetic resonance imaging (fMRI) during a verbal and a non-verbal auditory discrimination task. The stimuli were presented block-wise at three different intensities: 95, 85 and 75 dB (SPL). All subjects showed fMRI signal increases in superior temporal gyrus (STG) covering primary and secondary auditory cortex. Most importantly, the spatial extent of the fMRI response in STG increased with increasing stimulus intensity. It is hypothesized that spreading of excitation is associated with the encoding of increasing stimulus intensity levels. In addition, we found bifrontal activation supposedly evoked by the auditory articulary loop of working memory. The results presented here should assist in the design of optimal activation strategies for studying the auditory cortex with fMRI paradigms and may help in understanding intensity coding of auditory stimuli. PMID- 9740362 TI - The antisaccade: a review of basic research and clinical studies. AB - The ability to suppress reflexive responses in favor of voluntary motor acts is crucial for everyday life. Both abilities can be tested with an oculomotor task, the antisaccade task. This task requires subjects to suppress a reflexive prosaccade to a flashed visual stimulus and instead to generate a voluntary saccade to the opposite side. This article reviews what is currently known about the neural structures and processes which are involved in the performance of this task. Current data show that a variety of brain lesions, neurological diseases and psychiatric disorders result in errors, i.e. prosaccades towards the stimulus, in this task. Brain imaging studies have shown that a widely distributed cortical and subcortical network is active during the generation of antisaccades. These findings are discussed and the potential of the antisaccade task for diagnostic purposes is evaluated. PMID- 9740363 TI - Memory impairments associated with hippocampal versus parahippocampal-gyrus atrophy: an MR volumetry study in Alzheimer's disease. AB - Delayed memory impairments and medial temporal-lobe atrophy are considered to be cardinal features of Alzheimer's disease (AD). The goal of the present magnetic resonance (MR) volumetry study was to investigate the relationship between both features. We determined MR-derived estimates of hippocampal and parahippocampal volume in a sample of 27 AD patients and in a group of 26 healthy control subjects (NCs) of comparable age and education. We examined the performance of the two groups on immediate and delayed recall trials of an auditory-verbal list learning task (CVLT), a visual non-verbal memory task (Visual Reproduction of the WMS-R), and a screening procedure that provides an estimate of overall cognitive functioning (DRS). Volumes of the hippocampus and the parahippocampal gyrus were significantly smaller in AD patients than in NCs. AD patients were impaired in their overall level of cognitive functioning and showed memory deficits under immediate and delayed recall conditions. The association between medial temporal lobe atrophy and cognitive impairments in AD was found to be highly specific: Hippocampal volume correlated positively with delayed but not immediate recall of the verbal auditory list learning task. In contrast, parahippocampal gyrus volume, specifically in the right hemisphere, was positively related to delayed but not immediate recall of the non-verbal visual memory task. In NCs, there was a trend towards a negative association between hippocampal volumes and delayed verbal recall. Our results suggest that hippocampal and parahippocampal gyrus atrophy in AD are related to distinct aspects of the patients' memory impairments. Our findings have implications for current discussions regarding contributions of the hippocampus and the parahippocampal gyrus to memory in the intact human brain. PMID- 9740364 TI - Test/re-test reliability of the CANTAB and ISPOCD neuropsychological batteries: theoretical and practical issues. Cambridge Neuropsychological Test Automated Battery. International Study of Post-Operative Cognitive Dysfunction. AB - Neuropsychological test batteries are repeatedly administered to evaluate changes over time or the effects of clinical interventions. Relationships between scores on different tests within batteries are also examined to test models for associations between functional deficits. These comparisons may be misleading unless Test/Re-test reliability for individual tests is satisfactory. Interpretations of repeated measurements also depend on the extent to which improvement with practice varies between tasks and between more and less able individuals. Test/Re-test correlations and practice effects for two neuropsychological test batteries (CANTAB, ISPOCD) and from laboratory tasks commonly used in cognitive assessments of older people were obtained from large groups of healthy elderly. Tests in neuropsychological batteries varied markedly in test/re-test reliability which, in some cases, fell below levels considered methodologically acceptable. Putative measures of 'frontal' or 'executive' function, in which performance may be markedly improved by abrupt discovery of an appropriate strategy, were especially likely to show low reliability. Most tests showed significant practice effects, and on some these are substantial enough to compromise comparisons on repeated testing. On a minority of tests practice effects were counter-intuitive, in that less able showed significantly more gains than more able individuals. PMID- 9740365 TI - Interhemispheric transmission times in the presence and absence of the forebrain commissures: effects of luminance and equiluminance. AB - One subject (L.B.) with full forebrain commissurotomy, one (R.B.) with callosal agenesis and 20 normal controls were tested for simple reaction time (RT) with each hand, to visual stimuli in one or the other visual field. RTs for uncrossed conditions (hand ipsilateral to the visual field) were subtracted from RT to crossed conditions (hand contralateral to the visual field) to yield the crossed uncrossed difference (CUD), taken to be a measure of interhemispheric transfer time. CUDs increased from an average of 4.9 ms among the control subjects, to 23.3 ms for R.B., to 53.1 ms for L.B. Although overall RTs in all subjects increased with decreasing luminance of the stimuli, the CUD was not systematically affected and remained largely unaffected even under equiluminance. The results support previous evidence that interhemispheric transfer, even in the split brain, depends on visually insensitive pathways. PMID- 9740366 TI - Intact temporal memory in amnesic patients. AB - Current theories propose that amnesia is caused by an inability to encode the temporal properties of recent events and/or to associate information across time. The present investigation tested this postulation by manipulating the recency effect which is theorized to be caused by the encoding of temporal information. The continual-distractor paradigm was used to vary the temporal properties of recently presented lists. Amnesics' recall responded normally to the temporal manipulations in lists ranging from 18-54 s. In contrast, overall recall was impaired compared to normals in all conditions and across all positions, including the final position. These findings dissociate memory for temporal information from overall levels of recall. They suggest that the amnesic patient's memory deficit is not caused by an inability to encode temporally associated information. PMID- 9740367 TI - Age at onset and language disturbances in Alzheimer's disease. AB - This study examined the effect of age at symptom onset of Alzheimer's disease (AD) on the pattern of language disturbance. We assessed 150 consecutive patients with a clinical diagnosis of mild-to-moderate AD using the Western Aphasia Battery and a 100-item picture-naming test. A multivariate linear regression analysis examined the effect of age at onset after controlling for gender, education, severity of dementia and duration of the disease. Patients with early onset performed significantly worse than did patients with late onset on the word comprehension and sequential commands subtests. On the other hand, late-onset patients performed more poorly than early-onset patients on the picture-naming test in a subgroup with mild language deficits. However, the trend disappeared in other subgroups with more degraded language function. We consider that the concomitant effects of normal aging worsened the picture-naming deficits in the late-onset patients, and the rapid decline of naming ability in the early-onset patients masked the aging effect with the progression of language deficits. The deterioration of word comprehension and the rapid decline of naming ability are the characteristics of early-onset patients. The different patterns of language deficits between early- and late-onset patients may correspond to the genetic heterogeneity of AD. PMID- 9740368 TI - Indication of a common ancestry for copper tyrosinases and heme catalases revealed by hydrophobic cluster analysis of the brown locus protein sequence. PMID- 9740369 TI - Typical interaction patterns in alphabeta and betaalpha turn motifs. AB - A fully automatic classification procedure of short protein fragments is applied to identify connections between alpha-helices and beta-strands in a dataset of 141 protein chains. It yields 15 structural families of alphabeta turns and 15 families of betaalpha turns with at least five members. The sequence and structural features of these turn motifs are analysed with the focus on the local interactions located at alpha-helix and beta-strand ends. This analysis reveals specific interaction patterns that occur frequently among the members of many of the identified turn motifs. For the beta-strands, novel patterns are identified at the strands' entry and exit; they involve side chain/side chain contacts and beta-turns, generally of type I or II. For the alpha-helices, the interaction patterns consist of several backbone/backbone or backbone/side chain hydrogen bonds and of hydrophobic contacts; they generalize the well known N-terminal capping and C-terminal Schellman motifs. The interaction patterns at both ends of alpha-helices and beta-strands are found to constitute favourable structure motifs with low amino acid sequence specificity; their possible stabilizing role is discussed. Finally, the robustness of our classification procedure and of the description of N- and C-cap interaction patterns is validated by repeating our analysis on a larger dataset of 381 protein chains and showing that the results are maintained. PMID- 9740370 TI - Domain structural class prediction. AB - The structural class of a protein domain can be approximately predicted according to its amino acid composition. However, can the prediction quality be improved by taking into account the coupling effect among different amino acid components? This question has evoked much controversy because completely different conclusions have been obtained by different investigators. To resolve such a perplexing problem, predictions by means of various algorithms were performed based on the SCOP database (Murzin et aL, 1995), which is more natural and reliable for the study of structural classes because it is based on evolutionary relationships and on the principles that govern their three-dimensional structure. The results obtained using both resubstitution and jackknife tests indicated that the overall rates of correct prediction by an algorithm incorporating the coupling effect among different amino acid components were significantly higher than those by the algorithms that did not include such an effect. A completely consistent conclusion was also obtained when tests were performed on two large independent testing datasets classified into four and seven structural classes, respectively. It is revealed through an analysis that the reasons for reaching the opposite conclusion are mainly due to (1) misclassifying structural classes according to a conceptually incorrect rule, (2) misapplying the component-coupled algorithm by ignoring some important factors and (3) misrepresenting structural classes with statistically insignificant training subsets. Clarification of these problems would be instructive for effectively using the prediction algorithm and correctly interpreting the results. PMID- 9740371 TI - De novo design of a peptide which partitions between water and phospholipid bilayers as a monomeric alpha-helix. AB - To dissect the determinants of protein insertion into membranes, we designed a model peptide which partitions between water and phospholipid bilayers as an alpha-helical monomer. We used a simplex method to optimize the 'a, d hydrophobicity' and 'e, g charge' of a series of five peptides, where 'abcdefg' correspond to the positions in two turns of an alpha-helix. Circular dichroism and analytical ultra-centrifugation experiments showed that the final peptide (helix5) is monomeric and has an alpha-helix content of approximately 89% at 0 degrees C in aqueous solution. In the presence of large unilamellar vesicles (LUVs), helix5 partitions between the aqueous and membranous phases with a partition constant well suited for measurements by electron paramagnetic resonance (EPR) spectroscopy. EPR power saturation experiments with a cysteine scanning strategy showed that the alpha-helicity of helix5 is conserved upon binding to LUVs and that the alpha-helix binds parallel to the membrane surface with the central axis approximately 5 A below the lipid phosphate groups. Helix5 should be a useful model peptide for studies aimed at dissecting the determinants of the membrane binding of alpha-helices. The simplex-based strategy may be useful in the rational design of proteins when desired structural or partitioning properties cannot be selected or screened from libraries. PMID- 9740372 TI - Truncation and heme pocket mutations reduce production of functional catalase HPII in Escherichia coli. AB - The subunit of catalase HPII from Escherichia coli is 753 residues in length and contains a core of approximately 500 residues, with high structural similarity to all other heme catalases. To this core are added extensions of approximately 80 and 180 residues at the N- and C-termini, respectively. The tetrameric structure is made up of a pair of interwoven dimers in which 90 N-terminal residues of each subunit are inserted through a loop formed by the hinge region linking the beta barrel and alpha-helical domains of the adjacent subunit. A high concentration of proline residues is found in the vicinity of the overlap regions. To study the influence of the extended regions on folding and subunit association of HPII, a diversity of modifications have been introduced. Removal of the complete C terminal domain or the N-terminal extension, either separately or together, effectively creating a small subunit catalase, resulted in no enzyme accumulation. Systematic truncations showed that only nine C-terminal residues (Ile745 to Ala753) could be removed without significantly affecting the accumulation of active enzyme. Removal or even conservative replacements of the side chain of Arg744 significantly reduced the accumulation of active enzyme despite this residue interacting only with the C-terminal domain. Removal of as few as 18 residues from the N-terminus also reduced accumulation of active enzyme. Changes to other residues in the protein, including residues in the heme binding pocket, also reduced the accumulation of active protein without substantially affecting the enzyme specific activity. Implications of these data for the interdependence of subunit folding and subunit-subunit interactions are discussed. PMID- 9740373 TI - Studies on enzymatic activity and conformational stability of muscle acylphosphatase mutated at conserved lysine residues. AB - An oligonucleotide-directed mutagenesis study was carried out on the five acylphosphatase conserved lysine residues to assess their possible participation in enzyme active site formation and their contribution to the enzyme conformational stability. The study was designed to eliminate the ambiguity arising from the presence of a sulfate ion, an enzyme competitive inhibitor, bound to lysine 32 and 68 in the crystal structure of the erythrocyte isoenzyme. Furthermore, previous kinetic studies suggested the presence of residues with pKa=7.9 and 11, tentatively identified as two lysines. The kinetic parameters for the mutants under investigation are not significantly different from those of the wild-type enzyme, demonstrating that none of the lysine residues are involved in catalysis or in substrate binding. In addition, thermal and urea denaturation experiments performed by circular dichroism indicate that the mutated lysine residues do not play a significant role in the enzyme structural stabilization, as the destabilizing energy averages 1.40 kJ/mol. Such results are in agreement with those obtained with other proteins indicating that lysine residues make little contribution to the stability of the native structure. PMID- 9740374 TI - Mutation of cis-proline 207 in mitochondrial creatine kinase to alanine leads to increased acid stability. AB - We show that the mutation of an uncharged residue far from the active site to another uncharged residue can have effects on the active site without disturbing the overall structure of the protein. Cis-proline 207 of mitochondrial creatine kinase was mutated to alanine. The mutant showed a decrease in the pH-optimum for ATP synthesis by 1.5 units while the maximum relative activity was lowered to 53% of the wild-type enzyme. In the direction of ATP consumption, the pH optimum was lowered by 1.3 units and the maximum relative activity was 49% of the wild-type enzyme. The enzyme kinetic parameters Km and Kd for the substrates did not change dramatically, indicating a largely unperturbed active site. Small-angle X-ray scattering was used to investigate the structural change concomitant with the mutation, yielding a scattering profile only slightly different from that of the wild-type enzyme. Neither the radius of gyration nor the molecular mass showed any significant differences, leading to the conclusion that quarternary organization and fold of the mutant and the wild-type enzymes were similar. Theoretical analysis suggests the most probable primary source of structural change to be a transition of residue 207 peptide bond torsional angle co from the cis to the trans configuration. PMID- 9740375 TI - Use of protein engineering to explore subunit interactions in an allosteric enzyme: construction of inter-subunit hybrids in Clostridium symbiosum glutamate dehydrogenase. AB - Hybrids of different forms of clostridial glutamate dehydrogenase (GDH) have been constructed in order to probe the basis of allosteric interaction in this hexameric enzyme. It was shown that the C320S mutant, which is fully active and shows allosteric behaviour similar to that of the wild-type enzyme, can also be renatured after unfolding in urea. Mixtures of unfolded wild-type and C320S subunits gave rise to hybrids upon refolding. A purely random reassembly would lead to a simple binomial distribution. However there was a slightly better overall recovery of wild-type subunits and there appears to be a tendency for rapidly formed structured wild-type subunits in a mixture to nucleate further refolding in a way that biases the final distribution against the formation of C320S hexamers. Only the wild-type subunits in such hybrid mixtures are able to react with Ellman's reagent, 5,5'-dithiobis-(2-nitrobenzoate) (DTNB). Accordingly, after modification of hybrid hexamers with DTNB only the mutant subunits can bind NAD+. This permits fractionation on an NAD+-agarose affinity column. The elution pattern in itself indicates cooperativity since DTNB modification of just one subunit in a 1:5 wild-type/C320S hybrid largely abolished binding to the column. Kinetic studies were carried out on a fractionated preparation in which hexamers containing only one C320S subunit and five wild-type subunits were the predominant active species. Measurements of activity were made both before and after treatment with an excess of beta mercaptoethanol to remove the blocking thionitrobenzoate moieties. Before beta mercaptoethanol treatment this sample, with only one active subunit per hexamer, gave strictly hyperbolic (Michaelis-Menten) kinetics with NAD+ at pH 7.0, whereas after beta-mercaptoethanol (all six subunits now active) the markedly kinked Eadie-Hofstee plot characteristic of wild-type enzyme was obtained. On the other hand the sigmoid response to glutamate at high pH persisted (Hill coefficient=3.6) even without beta-mercaptoethanol, reflecting the fact that the inactive subunits can still bind glutamate. Beta-mercaptoethanol treatment restored full positive cooperativity (Hill coefficient=5.2). These results prove beyond doubt that the non-classical kinetic behaviour of clostridial GDH is a direct result of interaction between NAD+ binding sites on the six (normally) identical subunits of a hexamer. PMID- 9740376 TI - The affinity of cholera toxin for Ni2+ ion. AB - Cholera toxin (CT) was shown to bind to immobilized Ni2+ ion. The affinity of CT for the complex required the presence of the Ni2+ ion, since CT was unable to bind in its absence. Binding was mediated by the B-subunit (CTB) as both CT and CTB bound to the resin, but not the A-subunit (CTA). Binding was reversible in the presence of imidazole and suggested that the affinity of CT for the Ni2+ ion was mediated by His residues. The heat-labile enterotoxin of Escherichia coli (LT), which is closely related to CT, was unable to bind to the Ni2+ ion. Comparison of amino acid sequences revealed the presence of three His residues in CT (positions 13, 57 and 94), but only one in LT (position 57). To confirm that the residues at positions 13 and 94 of CTB were responsible for the binding, they were changed to residues found in LTB. Changing His13-->Arg completely abrogated the ability of CTB to bind to Ni2+ ion. In contrast, the mutation of His 94-->Asn reduced, but did not abrogate, the ability of CTB to bind to Ni2+ ion. Based on calculated interatomic distances, it is unlikely that His13 and His94 are part of the same complex. There appear to be two separate binding sites, with the principal site involving His13 and a much weaker site involving His94. This latter site can only participate in binding if the complex involving His13 has formed. PMID- 9740377 TI - Correct assembly of human normal adult hemoglobin when expressed in transgenic swine: chemical, conformational and functional equivalence with the human-derived protein. AB - Structural and functional investigations of recombinant human hemoglobin A (HbA) isolated from the erythrocytes of transgenic swine coexpressing human alpha- and beta-globins have been carried out to authenticate its correct expression, post translational processing and assembly. The HbA expressed in transgenic swine (TgHbA) is indistinguishable from the human-derived HbA in terms of its isoelectric pH, mass and elution pattern on a Mono S column. The chemical identity of the alpha- and beta-globin chains of TgHbA with the corresponding chains from human-derived HbA has been established by tryptic peptide mapping and amino acid sequencing. The proton NMR spectra of TgHbA have demonstrated that the conformational aspects of the protein around the heme pocket are indistinguishable from those of the control sample of HbA. The equivalence of the hydrogen bond pattern of TgHbA (in particular the inter-subunit surfaces) with that of authentic HbA has also been established by NMR studies. Consistent with these structural and conformational analyses, the TgHbA also exhibits complete functional equivalence with the human-derived HbA with respect to oxygen affinity, cooperativity, Bohr effect and allostery. Hence the studies presented here demonstrate that the transgenic swine system correctly transcribes the alpha and beta-globin transgenes, translates the respective alpha- and beta-globin mRNA to generate the corresponding globin chains, carries out the correct cotranslational processing of the translated globin chains, inserts the heme into the globin chains in the same orientation as in the human-derived HbA and assembles the alpha- and beta-subunits into a functionally cooperative tetramer that exhibits a response to allosteric effectors identical with that of human derived HbA. Thus, in the transgenic swine system, in vitro chemical manipulation steps such as those needed in the Escherichia coli and the yeast systems, to convert the rHbA expressed in these systems into forms functionally identical with that of the human-derived protein, are not needed. An additional advantage of the transgenic swine system is the stability of the transgenes over many generations. Hence the transgenic swine could serve as an excellent system for the production of human HbA (or its variants) for structure-function studies and for therapeutic applications. PMID- 9740378 TI - Structural and dynamic effects of point mutations in the recognition helix of the glucocorticoid receptor DNA-binding domain. AB - We have studied the wild type and two variants of the glucocorticoid receptor DNA binding domain (GRDBD): in one variant the three residues (the 'P-box' in GRDBD) that are essential for the discrimination between GREH and EREH are mutated to those in the estrogen receptor DBD (GRDBDega) and the other variant is a point mutation of one P-box residue, Ser459Gly (GRDBDggv). Molecular dynamics simulations (0.5-0.7 ns) have been performed on the GRDBDs, free in solution as well as in complex with the half-site response elements of the glucocorticoid (GREH) and estrogen (EREH) receptors. The residues which are central when forming the protein dimer interface in GRE-(GRDBD)2 (the 'D-box') were found to have different conformations in the different GRDBD-DNA complexes. This is consistent with experimental results showing that the cooperativity of dimeric GRDBD binding to DNA strongly depends on both the response element and the P-box residues. In our simulations the structures of GREH-GRDBDgsv (i.e. wild-type) and GREH GRDBDggv were more similar to each other than to the respective GRDBDs bound to EREH. This is due to a thymine methyl group which is present in the major groove of the GREH and prevents the first zinc coordinating subdomain in GRDBD to approach GREH, but which is absent in EREH. Thus, EREH-GRDBD is able to respond more to the Ser459Gly mutation than GREH-GRDBD. PMID- 9740379 TI - Improved folding of apo-retinol-binding protein in the periplasm of Escherichia coli: positive influences of dsbC coexpression and of an amino acid exchange in the vitamin A binding site. AB - The in vivo folding of the serum retinol-binding protein (RBP), a representative of the lipocalin structural family, is known to be complex. In order to gain insight into the essential steps along its folding pathway the heterologous production of the functional protein in Escherichia coli was investigated. Simultaneous overexpression of the bacterial dsbC gene, which codes for a periplasmic thiol-disulphide oxidoreductase, prevented the formation of soluble RBP variants with non-native disulphide bonds that were otherwise observed. Although the coexpression of dsbC had furthermore a stabilizing effect on the cell viability, the relative yield of the solubly produced RBP was not much better. In an attempt to enhance its folding efficiency, a favourable point mutation in the inner part of the retinol-binding pocket was predicted. Replacement of the polar Gln117 with an lie side chain seemed not only to relieve the unfavourable energetics of the carboxamide group in the environment of predominantly non-polar residues but also to fill an adjacent cavity in the hydrophobic core. Indeed, this single substitution reproducibly resulted in a more than threefold increase in the amount of functional recombinant RBP. Ligand binding experiments showed that the affinity of this mutant for retinol was slightly enhanced. Kinetic measurements revealed that this was due to a higher association rate whereas the dissociation of the complex with retinol was essentially unaffected. Although the question remained why nature did not select this obviously beneficial mutation, our results demonstrate that the folding pathway of a lipocalin can be optimized by protein engineering. PMID- 9740380 TI - Interventional ultrasound in Europe. AB - "Interventional ultrasound," defined as any diagnostic and therapeutic procedure performed under ultrasound guidance was first introduced in Europe, where its early development took place in Vienna, in Copenhagen, in Italy and in Switzerland. However, many of the applications of interventional ultrasound have been based on important pioneer work using other less-suitable needle guiding methods from the pre-ultrasound era. The European contributions to "interventional ultrasound" have especially been in the development of new puncture equipment, in the dissemination of various biopsy techniques, and draining procedures and, more recently, in the development of many different tissue-ablation techniques. The above contributions, which are outlined in this historical review, have, together with significant contributions from the rest of the world-not least from the United States and Japan-created a most efficient diagnostic as well as therapeutic tool for the benefit of our patients. PMID- 9740381 TI - Artifacts in intravascular ultrasound imaging during coronary artery stent implantation. AB - Intravascular ultrasound imaging is able to provide direct images of the stent meshwork. However, a paradoxical question remains unanswered: Why is it not possible to correct or prevent implantation defects by ultrasound-guided implantation? We postulate that these discrepancies are due to image artifacts. We performed an in vitro experiment allowing detection, physical characterization, and computerized simulations of the various aspects of these artifacts. The width of the echo of a strut is variable, dependent on its distance from the transducer. The stent strut echo orientation is variable, and depends on the position of the transducer inside the stent. The stent contour image depends on the position of the transducer. In conclusion, knowledge of these stent intravascular ultrasound image artifacts enabled us to discriminate accurately between artifacts and real stent implantation defects, and are indispensable for accurate qualitative and quantitative analyses of stents. PMID- 9740382 TI - Bladder filling reduces femoral artery wall distension and strain: beware of a full bladder! AB - During a previous study, we noted that the distension and strain of the femoral artery were relatively low when the bladder was full, a situation normally necessary for transabdominal echography. Therefore, in the present study we investigated the influence of bladder filling, if any, on wall properties of the common femoral artery. The results obtained were compared with those obtained in the common carotid artery. The study was performed on the right common carotid and right common femoral arteries of normotensive young (18-35 y) female volunteers (n = 24). Using a specially designed ultrasonic wall-tracking device and automatic brachial artery cuff blood pressure measurements, arterial distension (absolute change in diameter during the cardiac cycle; deltaD), strain (deltaD/D), and cross-sectional distensibility (DC) and compliance (CC) were determined before and after voiding. Distension and strain of the common femoral artery were significantly lower for a full than for an empty bladder. DC and CC were lower when the bladder was filled, but these differences did not reach the level of significance. Blood pressure as measured at the level of the brachial artery and heart rate were not statistically significantly different during a full or an empty bladder. It is concluded that bladder filling affects femoral artery wall properties, an observation that should be kept in mind when performing studies on artery wall properties at this level of the circulation. PMID- 9740383 TI - Ultrasonic mapping of the microvasculature: signal alignment. AB - The ultimate goal of this work was the development of a system capable of estimating the low flow velocities in the microvasculature. Estimation of low velocity flow within these vessels is challenging due to the small signal levels and the effect of cardiac and respiratory motion. Realignment of the signal from a single line-of-sight to remove physiological tissue motion is a critical part of the process of small-vessel flow mapping, and our methods for this alignment are considered in this paper. Each method involves the correlation of pulses acquired from the same line-of-sight. The first method involves the correlation of adjacent pulses (nearest-neighbor), the second involves a single reference line and the third involves averaging the correlation over a set of reference lines. We find that a nearest-neighbor strategy is suboptimal, and that strategies involving a global reference line are superior. A bound on the variance of estimates of the location of the correlation peak is presented. This bound allows us to consider our results in comparison with an absolute limit. Finally, a new algorithm allowing for alignment between lines-of-sight is described, and initial results are presented. Such an algorithm does, in fact, reduce jitter, correct for tissue motion and enables us to better visualize vessel continuity. We find that vessels as small as 40 microm can be mapped in two dimensions using a 50-MHz transducer. PMID- 9740384 TI - Variance components analysis of carotid and femoral intima-media thickness measurements. REGRESS Study Group, Interuniversity Cardiology Institute of The Netherlands, Utrecht, The Netherlands. Regression Growth Evaluation Statin Study. AB - B-mode ultrasound intima-media thickness (IMT) measurements of carotid and femoral arterial walls are used in atherosclerosis studies. In this study, the components contributing to IMT measurement variability in males with coronary artery disease were investigated by means of repeated B-mode ultrasound scans and repeated off-line video image analyses. For statistical analysis, a mixed-model analysis of variance was used. From sonographer data, it was shown that human subjects and their arterial wall segments contributed 75% of the total IMT measurement variability in this population. Inter-sonographer variance contributed 25%. The intra-sonographer variance was negligible (<1%). In off-line image analysis, variance components due to subjects and segments, inter-analyst variance, and residual fluctuation were 88%, < 1% and 11%, respectively. Intra analyst variance was negligible (<1%). The major source of B-mode ultrasound IMT measurement variability finds its origin in subjects and their arterial walls. Although sonographers proved a lesser source of variability, in comparative studies they should enter a study well trained and should be randomly assigned to subjects. Follow-up examinations should preferably be done by the same sonographer. Off-line image analysis contributed little to IMT measurement variability. PMID- 9740385 TI - Ultrasound imaging for arterial wall thickness measurement: an in vitro study with stereomicroscopic correlation. AB - Accuracy of B-mode ultrasonography was assessed in determining the arterial intima + media thicknesses at the near and far wall positions. From 10 human cadavers, 20 common carotid and 20 common femoral arteries were removed at autopsy and the intima + media thicknesses were measured with ultrasonography at the near and far wall locations and with stereomicroscopy. The differences between the locations were not statistically significant. The difference (mean +/ SD) between the stereomicroscopic and ultrasound measurements in the carotid artery was 0.12 +/- 0.09 mm at the near wall and 0.13 +/- 0.16 mm at the far wall and, in the femoral artery, 0.08 +/- 0.12 mm and 0.13 +/- 0.14 at the respective locations. At all locations, the stereomicroscope measurement was statistically significantly (p < 0.05) greater than the ultrasound measurement. PMID- 9740386 TI - Automatic registration of 3-D ultrasound images. AB - One of the most promising applications of 3-D ultrasound (US) lies in the visualisation and volume estimation of internal 3-D structures. Unfortunately, artifacts and speckle make automatic analysis of the 3-D data sets difficult. In this study, we investigated the use of 3-D spatial compounding to improve data quality, and found that precise registration is the key. A correlation-based registration technique was applied to 3-D ultrasound data sets acquired from in vivo examinations of a human gall bladder. We found that the registration technique performed well, and visualisation and segmentation of the compounded data were clearly improved. We also demonstrated that an automatic volume estimate made from the compounded data (13.0 mL) was comparable to a labour intensive manual estimate (12.5 mL). In comparison, automatic estimates of uncompounded data are less accurate (ranging from 13.5 mL to 16.7 mL). The registration technique also has applications in intra- and interpatient comparative studies. PMID- 9740387 TI - Rapid calibration for 3-D freehand ultrasound. AB - 3-D freehand ultrasound is a new imaging technique that is rapidly finding clinical applications. A position-sensing device is attached to a conventional ultrasound probe so that, as B-scans are acquired, they can be labelled with their relative positions and orientations. This allows a 3-D data set to be constructed from the B-scans. A key requirement of all freehand imaging systems is calibration; that is, determining the position and orientation of the B-scan with respect to the position sensor. This is typically a lengthy and tedious process that may need repeating every time a sensor is mounted on a probe. This paper describes a new calibration technique that takes only a few minutes to perform and produces results that compare favourably (in terms of both accuracy and precision) with previously published alternatives. PMID- 9740388 TI - Quantification of the enhanced backscatter phenomenon from an intravenous and an intra-arterial contrast agent. AB - The phenomenon of enhanced backscatter from myocardial contrast agents was studied using two examples, a robust thicker-walled, intra-arterial agent (AIP 201) and a smaller thinner-walled, intravenous agent (Quantison). Both agents are composed of albumin-encapsulated microbubbles. Samples of the agents were inserted into an in vitro phantom and insonated under different scanning regimes. Upon insonation, Quantison exhibited a pronounced increase in mean backscatter at medium and low concentrations, which decreased dramatically with increasing number of frames of insonation. At high concentrations, no dramatic decrease or increase in mean backscatter was observed over the period of the experiment. AIP 201 exhibited an overall decrease in mean backscatter with increasing number of frames of insonation. These results suggest that the difference in size and wall thickness of the contrast microcapsules can significantly affect the behaviour of the contrast agents in an ultrasound field. PMID- 9740389 TI - Aliasing-tolerant color Doppler quantification of regurgitant jets. AB - Conservation of momentum transfer in regurgitant cardiac jets can be used to calculate the flow rate from color Doppler velocities. In this study, turbulent jets were simulated by finite elements; pseudocolor Doppler images were interpolated from the computations, with aliasing introduced artificially. Jets were also imaged by color Doppler in an in vitro flow system. To suppress aliasing errors, jet velocities were fitted iteratively to a fluid mechanical model constrained to match the orifice velocity (measured without aliasing by continuous-wave Doppler). At each iteration, the model was used to detect aliased velocities, which were excluded during the next iteration. Iteration continued until the flow rate calculated by the model and number of calculated nonaliased pixels were unchanged. The good correlations between measured and calculated flow rates in the experimental (R2 = 0.933) and computational studies (R2 = 0.990) suggest that this may be a clinically useful approach even in aliased images. Published by Elsevier Science Inc. PMID- 9740390 TI - Factors affecting color Doppler energy ultrasound recordings in an in-vitro model. AB - Compared to conventional color Doppler ultrasound imaging, the new color Doppler modality "color Doppler energy" (CDE) has improved the possibility of visualizing blood vessels having low blood-flow velocities, but appears to be influenced by the settings of the ultrasound instrument and motion artefacts. The aim of this methodological study was to evaluate the effects of the different factors on the CDE signal. The CDE mode of a commercially available ultrasound system (Acuson 128 XP) was tested in an in vitro study. The effect of depth, angle of insonation, flow velocity, instrument power output, gain and other instrument settings were evaluated. The CDE signals obtained were stored on videotape and subsequently subjected to off-line computer analysis. The CDE signal intensity was found to be influenced mainly by fluid flow velocity, but was also affected by depth and instrument settings. Gain and power had, however, limited influence in this setting. Thus, the intensity of the CDE signal is influenced by several factors. Our results emphasize the need for optimum fixed preinstalled instrument settings when attempting to quantify organ perfusion by use of this new technique. PMID- 9740391 TI - Hemostasis of punctured blood vessels using high-intensity focused ultrasound. AB - The hemorrhagic complications of vascular injury can be significant. We report on the use of high-intensity focused ultrasound (HIFU) to stop the hemorrhage of punctured blood vessels in pigs. Two HIFU transducers with frequencies of 3.5 and 2.0 MHz, each equipped with a water-filled conical housing, were used. Major blood vessels (femoral artery and vein, axillary artery, carotid artery and jugular vein), 2-10 mm in diameter, of anesthetized pigs were exposed surgically and punctured with 14- and 18-gauge needles to produce moderate to profuse bleeding. Complete hemostasis was achieved in less than 3 min of HIFU treatment in most blood vessels, and all vessels were patent after the treatment. Both HIFU frequencies were effective in producing hemostasis. Gross examination of the HIFU treated vessels showed a consistent hardening of the soft tissue surrounding the blood vessels, providing a seal for the puncture hole. Microscopic examination of the vessels showed a remarkably localized HIFU treatment, resulting in coagulation of the adventitia, and an extensive fibrin network around the vessels and in the puncture hole. The vessel walls exhibited focal swelling, without evidence of irreversible injury. HIFU may provide a useful method for achieving hemostasis of punctured and traumatized blood vessels in a variety of clinical settings. PMID- 9740392 TI - An investigation of possible effects of high-frequency ultrasound on cellular integrity of cultured fibroblasts. AB - Several investigators have demonstrated the feasibility of imaging at the cellular level using acoustical microscopy. It has also been proposed that acoustical microscopy technology might be adopted for in vivo applications. Before such applications are implemented, it is important to demonstrate that any major deleterious effects are highly unlikely. To this end, we have repeatedly scanned NIH/3T3 mouse fibroblasts in culture using an Olympus UH3 acoustical microscope operating at 600 MHz. No adverse effects were observed even after exposures for 1 h. Spatial peak temporal averaged intensities were estimated to be below 300 mW/cm2. PMID- 9740393 TI - An analysis of elastographic contrast-to-noise ratio. AB - We present a theoretical formalism and simulation results that allow the incorporation of the elastic contrast properties of tissues with simple geometries into the elastographic noise models developed previously. This analysis results in the computation of the elastographic contrast-to-noise ratio (CNRe). The CNRe in elastography is an important quantity that is related to the detectability of a lesion or inhomogeneity. In this paper, the upper bound on the elastographic CNRe is derived for both a one-dimensional (1-D) and 2-D analytic plane-strain tissue model. The CNRe in the elastogram depends on the contrast transfer efficiency (CTE) for both the 1-D and 2-D geometries discussed in this paper. The 1-D model is used to characterize layered structures and the 2-D model is derived for circular inclusion within a background of uniform elasticity. A previously derived classical analytic solution of the elasticity equations, for a circular inclusion embedded in an infinite medium and subjected to a uniaxial compression, is used to compute the upper bound of the CNRe. Monte Carlo simulations illustrate the close correspondence between the theoretical and simulation results. PMID- 9740394 TI - Cyclooxygenase-2 expression is increased in frontal cortex of Alzheimer's disease brain. AB - Many epidemiological studies suggest that use of nonsteroidal anti-inflammatory drugs delays or slows the clinical expression of Alzheimer's disease, but the mechanism by which these drugs might affect pathophysiological processes relevant to Alzheimer's disease has been unclear. Non-steroidal anti-inflammatory drugs are presumed to act by inhibiting cyclooxygenase, a key enzyme in the metabolism of membrane-derived arachidonic acid into prostaglandins. In recent years, two distinct isoforms of cyclooxygenase have been characterized, a constitutive form, cyclooxygenase-1, and a mitogen-inducible form, cyclooxygenase-2. Cyclooxygenase 2 has been identified in rodent brain. Excitotoxic lesions cause up-regulation of cyclooxygenase-2 expression coincident with the onset of expression of markers of apoptosis; cyclooxygenase-2 thus represents a possible target of non-steroidal anti-inflammatory drug action in neurodegenerative mechanisms. In the present study, we examined cyclooxygenase-2 gene expression in Alzheimer's disease and control cases. We found up-regulation of cyclooxygenase-2 expression in Alzheimer's disease frontal cortex. Further, we found that synthetic beta-amyloid peptides induced cyclooxygenase-2 expression in SH-SY5Y neuroblastoma cells in vitro, suggesting a mechanism for cyclooxygenase-2 up-regulation in Alzheimer's disease. These findings support the investigation of selective cyclooxygenase-2 inhibitors for the treatment of Alzheimer's disease. PMID- 9740395 TI - Characterization of human presenilin 1 transgenic rats: increased sensitivity to apoptosis in primary neuronal cultures. AB - Mutations in the gene for presenilin 1 are causative for the majority of cases of early onset familial Alzheimer's disease. Yet, the physiological function of presenilin 1 and the pathological mechanisms of the mutations leading to Alzheimer's disease are still unknown. To analyse potential pathological effects of presenilin 1 over-expression, we have generated transgenic rats which express high levels of human presenilin 1 protein in the brain. The over-expression of presenilin 1 leads to saturation of its normal processing and to the appearance of full-length protein in the transgenic rat brain. The transgenic protein is expressed throughout the brain and is predominantly found in neuronal cells. Cultured primary cortical neurons derived from these transgenic rats are significantly more sensitive than non-transgenic controls to apoptosis induced by standard culture conditions and to apoptosis induced by trophic factor withdrawal. Furthermore, the observed apoptosis is directly correlated with the expression of the transgenic protein. The results further emphasize the role of presenilin 1 in apoptotic cell death in native neuronal cultures. PMID- 9740396 TI - The role of apoptosis and excitotoxicity in the death of spinal motoneurons and interneurons after neonatal nerve injury. AB - There is evidence that motoneurons which die following neonatal nerve injury in rats do so through an excitotoxic mechanism. In this study, we have investigated whether this excitotoxicity induces motoneuron death by apoptosis. Sciatic motoneurons were prelabelled at birth with the retrograde tracing agent, Fast Blue, and the sciatic nerve was crushed in one leg two days later. At intervals up to 12 days, sections of the lumbar enlargement were analysed for apoptosis using propidium iodide and terminal deoxynucleotidyl transferase biotin-14-UTP nick end labelling techniques. A significant concentration of Fast Blue-labelled apoptotic motoneurons was seen in the area of the sciatic motor pool ipsilateral to the nerve injury, with the majority occurring in the first three days. Comparison of estimates of the time-course of apoptosis with that of motoneuron survival suggest that all motoneuron death induced during the first 12 days occurs by apoptosis and that the process is only recognizable for 2 h. Treatment with the N-methyl-D-aspartate receptor antagonist, dizocilpine maleate, reduced the level of apoptosis by 60%. Taken together, these data show that motoneurons which have been affected by an excitotoxic mechanism die by apoptosis. The apoptotic study also provides evidence, for the first time, that unilateral nerve injury induces motoneuron death in the contralateral sciatic motor pool. Apoptotic interneurons were also seen on both sides of the spinal cord as a result of nerve injury. PMID- 9740397 TI - Long-term changes in the aggregation state and toxic effects of beta-amyloid injected into the rat brain. AB - The long-term effects of beta-amyloid peptide 1-40 injection into the rat forebrain were studied. Ten micrograms of pre-aggregated peptide were injected into the right nucleus basalis of male Wistar rats which were then killed four or six months later. Congo Red staining of histological sections showed that the peptide deposit was aggregated in a fibrillary form four months post-surgery, whereas at six months almost no trace of birefringency was detected at the deposit site, indicating a loss of fibril organization. This result was confirmed by electron microscopic analysis of the peptide deposits. The presence of the peptide at the injection site six months post-surgery was demonstrated by both Haematoxylin staining and beta-amyloid immunoreactivity. The number of choline acetyltransferase-immunoreactive neurons was reduced by 66% in the injected nucleus basalis four months after injection. A decrease in cortical acetylcholine release was also found at this time. Concomitantly with the loss of fibril conformation, a complete recovery of choline acetyltransferase immunoreactivity in the nucleus basalis and of acetylcholine release in the cortex was observed at six months. These data provide in vivo evidence that beta-amyloid neurotoxicity is related to the fibrillary conformation of the peptide aggregates, thus confirming previous in vitro studies. PMID- 9740398 TI - Enduring cognitive, neurobehavioral and histopathological changes persist for up to one year following severe experimental brain injury in rats. AB - Clinical studies have demonstrated that patients sustain prolonged behavioral deficits following traumatic brain injury, in some cases culminating in the cognitive and histopathological hallmarks of Alzheimer's disease. However, few studies have examined the long-term consequences of experimental traumatic brain injury. In the present study, anesthetized male Sprague-Dawley rats (n = 185) were subjected to severe lateral fluid-percussion brain injury (n = 115) or sham injury (n = 70) and evaluated up to one year post-injury for cognitive and neurological deficits and histopathological changes. Compared with sham-injured controls, brain-injured animals showed a spatial learning impairment that persisted up to one year post-injury. In addition, deficits in specific neurologic motor function tasks also persisted up to one year post-injury. Immunohistochemistry using multiple antibodies to the amyloid precursor protein and/or amyloid precursor protein-like proteins revealed novel axonal degeneration in the striatum, corpus callosum and injured cortex up to one year post-injury and in the thalamus up to six months post-injury. Histologic evaluation of injured brains demonstrated a progressive expansion of the cortical cavity, enlargement of the lateral ventricles, deformation of the hippocampus, and thalamic calcifications. Taken together, these findings indicate that experimental traumatic brain injury can cause long-term cognitive and neurologic motor dysfunction accompanied by continuing neurodegeneration. PMID- 9740399 TI - Prolonged enhancement and depression of synaptic transmission in CA1 pyramidal neurons induced by transient forebrain ischemia in vivo. AB - Evoked postsynaptic potentials of CA1 pyramidal neurons in rat hippocampus were studied during 48 h after severe ischemic insult using in vivo intracellular recording and staining techniques. Postischemic CA1 neurons displayed one of three distinct response patterns following contralateral commissural stimulation. At early recirculation times (0-12 h) approximately 50% of neurons exhibited, in addition to the initial excitatory postsynaptic potential, a late depolarizing postsynaptic potential lasting for more than 100 ms. Application of dizocilpine maleate reduced the amplitude of late depolarizing postsynaptic potential by 60%. Other CA1 neurons recorded in this interval failed to develop late depolarizing postsynaptic potentials but showed a modest blunting of initial excitatory postsynaptic potentials (non-late depolarizing postsynaptic potential neuron). The proportion of recorded neurons with late depolarizing postsynaptic potential characteristics increased to more than 70% during 13-24 h after reperfusion. Beyond 24 h reperfusion, approximately 20% of CA neurons exhibited very small excitatory postsynaptic potentials even with maximal stimulus intensity. The slope of the initial excitatory postsynaptic potentials in late depolarizing postsynaptic potential neurons increased to approximately 150% of control values up to 12 h after reperfusion indicating a prolonged enhancement of synaptic transmission. In contrast, the slope of the initial excitatory postsynaptic potentials in non-late depolarizing postsynaptic potential neurons decreased to less than 50% of preischemic values up to 24 h after reperfusion indicating a prolonged depression of synaptic transmission. More late depolarizing postsynaptic potential neurons were located in the medial portion of CA1 zone where neurons are more vulnerable to ischemia whereas more non-late depolarizing postsynaptic potential neurons were located in the lateral portion of CA1 zone where neurons are more resistant to ischemia. The result from the present study suggests that late depolarizing postsynaptic potential and small excitatory postsynaptic potential neurons may be irreversibly injured while non-late depolarizing postsynaptic potential neurons may be those that survive the ischemic insult. Alterations of synaptic transmission may be associated with the pathogenesis of postischemic neuronal injury. PMID- 9740400 TI - Expression of alpha3, beta3 and gamma1 GABA(A) receptor subunit messenger RNAs in visual cortex and lateral geniculate nucleus of normal and monocularly deprived monkeys. AB - Complementary RNA probes derived from complementary DNA specifically subcloned from monkey tissue were used to localize, by in situ hybridization histochemistry, the relatively rare alpha3, beta3 and gamma1 subunit transcripts of the GABA(A) receptor in visual cortex and lateral geniculate nucleus of normal monkeys and in monkeys that had been deprived of vision in one eye. Overall, levels of alpha3, beta3 and gamma1 subunit transcripts were very low. In the primary visual cortex (area 17) they were concentrated in layers II and VI and in a stratum of white matter subjacent to layer VI. The localization and density of the three messenger RNAs closely resembled those of other rare (alpha2, alpha5 and beta1) transcripts but their distribution also overlapped that of the predominant alpha1, beta2 and gamma2 subunit transcripts. In area 18, alpha3 and beta3 transcript distribution resembled that in area 17, with the addition of a third band of hybridization in layer IV for beta3. Gamma1 subunit transcript localization in area 18 differed significantly from that in area 17, with increased expression restricted to layer IV. In the dorsal lateral geniculate nucleus, beta3 and gamma1 transcripts were expressed at low levels across all layers while alpha3 transcripts were restricted to the magnocellular layers. Following 15 and 18 day periods of monocular deprivation, induced by intravitreal injections of tetrodotoxin, levels of alpha3 receptor subunit transcripts showed modest reductions in layer VI of area 17 and in deprived geniculate laminae of adult animals. Reductions in alpha3 transcript levels were much more pronounced in layer IVCbeta of a five-month-old monkey deprived for the same time. Levels of beta3 and gamma1 transcripts were unaffected by monocular deprivation in cortex and geniculate at any age. Taken together with studies of other GABA(A) receptor transcripts, these results demonstrate the heterogeneity of GABA(A) receptor messenger RNA expression in the monkey geniculo-striate pathway and the varied response to reduced neuronal activity. PMID- 9740401 TI - Spatio-temporal diversity in the microenvironments for neural cell adhesion molecule, neural cell adhesion molecule-polysialic acid, and L1-cell adhesion molecule expression by sensory neurons and their targets during cochleo vestibular innervation. AB - Sixteen phases in the microenvironments were defined for the structural development and innervation of the cochleo-vestibular ganglion and its targets. In each phase the cell adhesion molecules, neural cell adhesion molecule, neural cell adhesion molecule-polysialic acid, and L1-cell adhesion molecule, were expressed differentially by cochleo-vestibular ganglion cells, their precursors, and the target cells on which they synapse. Detected by immunocytochemistry in staged chicken embryos, in the otocyst, neural cell adhesion molecule, but not L1 cell adhesion molecule, was localized to the ganglion and hair cell precursors. Ganglionic precursors, migrating from the otocyst, only weakly expressed neural cell adhesion molecule. Epithelial hair cell precursors, remaining in the otocyst, expressed neural cell adhesion molecule, but not L1-cell adhesion molecule. Post-migratory ganglion cell processes expressed both molecules in all stages. The cell adhesion molecules were most heavily expressed by axons penetrating the otic epithelium and accumulated in large amounts in the basal lamina. In the basilar papilla (cochlea), cell adhesion molecule expression followed the innervation gradient. Neural cell adhesion molecule and L1 were heavily concentrated on axonal endings peripherally and centrally. In the rhombencephalon, primitive epithelial cells expressed neural cell adhesion molecule, but not L1-cell adhesion molecule, except in the floorplate. The neuroblasts and their axons expressed L1-cell adhesion molecule, but not neural cell adhesion molecule, when they began to migrate and form the dorsal commissure. There was a stage-dependent, differential distribution of the cell adhesion molecules in the floorplate. Commissural axons expressed both cell adhesion molecules, but their polysialic acid disappeared within the floorplate at later stages. In conclusion, the cell adhesion molecules are expressed by the same cells at different times and places during their development. They are positioned to play different roles in migration, target penetration, and synapse formation by sensory neurons. A multiphasic model provides a morphological basis for experimental analyses of the molecules critical for the changing roles of the microenvironment in neuronal specification. PMID- 9740402 TI - Multiple roles of neural cell adhesion molecule, neural cell adhesion molecule polysialic acid, and L1 adhesion molecules during sensory innervation of the otic epithelium in vitro. AB - To explore the role of cell adhesion molecules in the innervation of the inner ear, antibody perturbation was used on histotypic co-cultures of the ganglionic and epithelial anlagen derived from the otocyst. When unperturbed, these tissues survived and differentiated in this culture system with outgrowth of fasciculated neuronal fibers which expressed neural cell adhesion molecule and L1. The fibers exhibited target choice and penetration, then branching and spreading within the otic epithelium as individual axons. Treatment of the co-cultures, or of the ganglionic anlagen alone, with anti-neural cell adhesion molecule or anti-L1 Fab fragments produced a defasciculation of fibers but did not affect neurite outgrowth. In the co-cultures this defasciculation was accompanied by a small increase in the number of fibers found in inappropriate tissues. However, the antibodies did not prevent fiber entry to the otic epithelium. In contrast, removal of polysialic acid from neural cell adhesion molecule with endoneuraminadase-N, while producing a similar fiber defasciculation, also increased the incidence of fibers entering the epithelium. Nevertheless, once within the target tissue, the individual fibers responded to either Fab or to desialylation by spreading out more rapidly, branching, and growing farther into the epithelium. The findings suggest that fasciculation is not essential for specific sensory fibers to seek out and penetrate the appropriate target, although it may improve their tracking efficiency. Polysialic acid on neural cell adhesion molecule appears to limit initial penetration of the target epithelium. Polysialic acid as well as neural cell adhesion molecule and L1 function are involved in fiber-target interactions that influence the arborization of sensory axons within the otic epithelium. PMID- 9740403 TI - Regulation of nitric oxide synthase messenger RNA expression in the rat hippocampus by glucocorticoids. AB - Nitric oxide and glucocorticoids have been implicated in learning and memory, as well as in regulation of the stress response. By use of the in situ hybridization technique, we examined the role of glucocorticoids in the regulation of nitric oxide synthase messenger RNA in the hippocampus. In control animals, nitric oxide synthase subtype I (neuronal) messenger RNA was expressed in the CA1, CA3 and dentate gyrus of the hippocampus. Nitric oxide synthase subtype I expression was almost absent in CA2 pyramidal neurons. Neither subtype II (immunological) nor subtype III (endothelial) nitric oxide synthase messenger RNAs were observed in neurons of the hippocampal subfields. Bilateral removal of the adrenal glands resulted in a significant increase in nitric oxide synthase subtype I messenger RNA expression in the CA1 and CA3 pyramidal neurons and in granular cells of the dentate gyrus. To a lesser degree, the nitric oxide synthase subtype I messenger RNA signal was increased in CA2 pyramidal neurons. Daily administration of glucocorticoids for one week attenuated the adrenalectomy-induced increased level of expression of the messenger RNA encoding nitric oxide synthase subtype I in all areas studied. Because adrenalectomy, which suppresses the production of glucocorticoids, increases nitric oxide synthase expression, and replacement of adrenalectomized animals with glucocorticoids restores the basal levels of nitric oxide synthase subtype I expression, our results demonstrate an up-regulation of nitric oxide synthase subtype I messenger RNA in the absence of glucocorticoids in the hippocampus. The present findings suggest an involvement of the stress axis in the regulation of the synaptic plasticity process mediated by nitric oxide in the hippocampus. PMID- 9740404 TI - Tyrosine kinase A, galanin and nitric oxide synthase within basal forebrain neurons in the rat. AB - Cholinergic basal forebrain neurons appear to play a key role in cognition and attention. In rat, basal forebrain neurons express multiple proteins including the high-affinity signal transducing tyrosine kinase A receptor for nerve growth factor, the neuropeptide galanin and nitric oxide synthase, a marker for the novel neurotransmitter nitric oxide. The present study was undertaken to define the relationship between neurons expressing each of these markers within the medial septum-vertical limb of the diagonal band, horizontal limb of the diagonal band and nucleus basalis in colchicine pre-treated rats. Tyrosine kinase A immunopositive neurons were seen throughout all subfields of the basal forebrain. In contrast, nitric oxide synthase- and galanin-immunoreactive neurons were mainly distributed within the septal-diagonal band complex. Co-localization experiments revealed that virtually all nitric oxide synthase-positive neurons (visualized by nicotinamide adenine dinucleotide phosphate-diaphorase histochemistry) also contained tyrosine kinase A, whereas many fewer tyrosine kinase A neurons were nicotinamide adenine dinucleotide phosphate-diaphorase positive within the medial septum-vertical limb of the diagonal band. Within the horizontal limb of the diagonal band, numerous nicotinamide adenine dinucleotide phosphate-diaphorase neurons expressed tyrosine kinase A, whereas only a small number of tyrosine kinase A neurons contained nicotinamide adenine dinucleotide phosphate-diaphorase. Within the nucleus basalis very few neurons were nicotinamide adenine dinucleotide phosphate-diaphorase reactive, and a minor number contained tyrosine kinase A. Additional co-localization experiments revealed minor percentages of neurons containing nicotinamide adenine dinucleotide phosphate-diaphorase and galanin immunoreactivity within the various subfields of the basal forebrain. Within the horizontal limb of the diagonal band minor numbers of nicotinamide adenine dinucleotide phosphate-diaphorase-reactive perikarya displayed galanin. Similarly, only a few galanin-containing neurons expressed nicotinamide adenine dinucleotide phosphate-diaphorase. The existence of tyrosine kinase A, nitric oxide synthase and galanin within select neuronal subgroups of the cholinergic basal forebrain suggests that these perikarya are responsive to a complex set of chemical signals. A greater understanding of the chemical signature of the cholinergic basal forebrain neurons will provide the insight required to develop novel pharmacological approaches aimed at preventing or slowing the degenerative processes that effect these neurons in aging and pathologic disorders. PMID- 9740405 TI - Fluoxetine induces the transcription of genes encoding c-fos, corticotropin releasing factor and its type 1 receptor in rat brain. AB - Fluoxetine is a serotonin re-uptake blocker commonly used to treat endogenous depression. The present experiments were carried out to assess the effects of fluoxetine on c-fos induction throughout the rat brain. In addition, intron directed in situ hybridization analysis was used to examine fluoxetine regulation of corticotropin-releasing factor heteronuclear gene transcription in the paraventricular nucleus of the hypothalamus. Because the actions of corticotropin releasing factor are mediated by membrane-bound corticotropin-releasing factor type 1 receptors, we also evaluated the stimulation of such receptors after acute fluoxetine exposure. The immediate-early gene, c-fos, was markedly induced in several telencephalic and diencephalic brain structures. For instance, a strong hybridized signal was apparent 30 min after fluoxetine (10 mg/kg; intraperitoneal) administration in the caudate putamen, septal nucleus, bed nucleus of stria terminalis, anterodorsal preoptic area, paraventricular nucleus, supraoptic nucleus, ventromedial hypothalamus and posterior hypothalamic nucleus. In addition, c-fos-expressing neurons were also evident in discrete amygdaloid nuclei. This nuclear induction was brief in duration, as levels of the immediate early gene were mostly undetectable 90 min after drug administration. In contrast to the extensive induction of c-fos by fluoxetine throughout the brain parenchyma, elevation of corticotropin-releasing factor heteronuclear RNA levels were confined exclusively to neurosecretory nerve cells of the paraventricular nucleus, with peak levels detected 30 min after fluoxetine exposure. Therefore, the time-course of corticotropin-releasing factor heteronuclear RNA closely paralleled that of c-fos. Significant changes in corticotropin-releasing factor type 1 receptor messenger RNA levels were also observed in the paraventricular nucleus but with a slow incremental biosynthesis of the receptor messenger RNA, as high levels were discernible only 360 min after fluoxetine treatment. Finally, we failed to detect sex-related differences in the acute response to fluoxetine, as both female and male rat brains showed a comparable induction of c-fos, corticotropin-releasing factor heteronuclear RNA and corticotropin-releasing factor type 1 receptor expression within parvocellular neurosecretory nerve cells that govern the stress response. All of these findings are discussed in terms of specific sequences of nuclear events that couple fluoxetine-based serotonin input with changes in gene expression in selective neurons. PMID- 9740406 TI - Behavioural characterization and amounts of brain monoamines and their metabolites in mice lacking histamine H1 receptors. AB - Behavioural assessments were made of mutant mice lacking histamine H1 receptors to reveal the function of H1 receptors in the behaviour of mice. Exploratory behaviour of mice in a new environment was examined to discover whether the absence of H1 receptors in mice affects actions relating to their emotions. The H1 receptor-deficient mice showed a significant decrease in ambulation in an open field and on an activity wheel. Cognitive functions and anxiety were examined using passive avoidance response test and the elevated plus-maze test, respectively. The passive avoidance test did not show any change in latency. The elevated plus-maze test revealed that the transfer latency of the mutant mice was significantly prolonged, indicating that H1 receptors are partly associated with the control of anxiety. Aggressive behaviour was examined by a resident-intruder aggression test. When confronted with an intruder, the mutant mice attacked the intruder significantly slower and less frequently than did wild-type mice after a six-month isolation period. A formalin test and a forced swimming test were used to evaluate the nociceptive response and depressive or despairing state, respectively, of both groups. The mutant mice showed a significant decrease of nociceptive response in the late phase without affecting the early phase. There was no significant difference in the forced swimming test between the two groups. The brain content of monoamines and their metabolites was measured in the H1 receptor null and wild-type mice. The turnover rate of 5-hydroxytryptamine defined by the ratio of 5-hydroxyindoleacetic acid and 5-hydroxytryptamine was significantly increased in the cerebral cortex and hippocampus of H1 receptor null mice. These results support the previous pharmacological findings that histamine modulates various neurophysiological functions such as locomotor activity, emotion, memory and learning, nociception and aggressive behaviour through H1 receptors. PMID- 9740407 TI - Regional and temporal separation of serotonergic activity mediating social stress. AB - Stressful aggressive interaction stimulates central serotonergic activation in telencephalon as well as brainstem. Social roles can be distinguished by monoamine activity following aggression. Pairs of male lizards, Anolis carolinensis, were allowed to fight and form dominant/subordinate relationships. In micropunched regions of telencephalon, the greatest serotonergic changes occur in subordinate males. In hippocampal cortex and nucleus accumbens, subordinate males have increased 5-hydroxyindoleacetic acid/serotonin at 1 h following the fight. In these areas the ratio gradually decreases over a week of cohabitation, as was previously reported for brainstem. Medial and lateral amygdala develop increased serotonergic activity more slowly, with the greatest increase being evident following a week of interaction. Turnover, serotonin and 5 hydroxyindoleacetic acid levels in amygdala escalate over the first week of interaction in subordinate males, and return to baseline by one month. In dominant males, the pattern is accelerated, with the most extensive serotonin system activity present at 1 h, then decreasing over a month. The patterns of serotonergic activation are so similar in hippocampus, nucleus accumbens and brainstem that a co-ordinated response may be involved in mediating short-term social stress and aggression. Similarly, medial and lateral amygdala exhibit corresponding, but delayed patterns in subordinate males, suggesting a co ordinated response in these regions mediating longer-term stress responses. These data are consistent with rapid neuroendocrine stress modulation in dominant individuals, and delayed serotonergic activity changes in subordinate males. PMID- 9740408 TI - Astrocytes are more vulnerable than neurons to cellular Ca2+ overload induced by a mitochondrial toxin, 3-nitropropionic acid. AB - The differential effects of 3-nitropropionic acid on cultured neurons and astrocytes (of cortical and striatal origin) were examined by studying intracellular Ca2+ changes using imaging techniques with fura-2. The neurons and astrocytes whose intracellular Ca2+ concentration was recorded were identified later by immunocytochemical staining for microtubule-associated protein 2 and glial fibrillary acidic protein, respectively. 3-Nitropropionic acid (1.7 mM) irreversibly increased intracellular Ca2+ in astrocytes (27%) and, to a significantly smaller extent, in neurons (10%). The latency to onset of the intracellular Ca2+ increase was longer in neurons (45 min) than in astrocytes (29 min). Thus, a differential susceptibility of astrocytes and neurons was observed. The 3-nitropropionic acid-induced astrocytic and neuronal Ca2+ accumulations were both due to influx of Ca2+, as the increases were absent in Ca2+-free medium. An inhibitor of the Na+-Ca2+ exchanger (2',4'-dichlorobenzamil), greatly reduced the intracellular Ca2+ increase in astrocytes, but not in neurons. This indicates that the intracellular Ca2+ increase in astrocytes is primarily mediated by a reverse operation of the Na+-Ca2+ exchange system, whereas in neurons it is mediated by a different mechanism. In addition, we noted that astrocytic cell death occurred in 9% of cells at 60 min or more after the start of a 40 min perfusion with 3-nitropropionic acid, while only 4% of neurons died. In astrocytes, cell death was preceded by blebbing of the cell membrane, and by a sustained increase in intracellular Ca2+ followed by an abrupt further elevation occurring just before cellular collapse. The present results indicate that astrocytes are more vulnerable than neurons to 3-nitropropionic acid-induced cellular Ca2+ overload and toxicity, and hence support the hypothesis that, in part, 3-nitropropionic acid-induced neurotoxicity could be secondary to astrocytic cell death caused by Ca2+ overload. PMID- 9740409 TI - Patch-clamp recordings of membrane currents evoked during natural synaptic activity in sympathetic neurons. AB - Membrane currents elicited by colonic distension and by electrical stimulation of the intermesenteric nerve containing colonic afferent nerve fibres were recorded from neurons of the mouse superior mesenteric ganglion at 20 degrees C with the whole-cell patch-clamp method. Electrically-evoked excitatory postsynaptic currents reversed at -3.5 mV. At membrane holding voltages of -70 mV and -110 mV, the excitatory postsynaptic currents were characterized by a single exponential decay with a mean (+/- S.E.M.) time-constant of 17.5 +/- 1.3 ms and 15.5 +/- 2.3 ms, respectively. Colonic distension evoked a series of the excitatory postsynaptic currents which ranged in amplitude from 10 to 700 pA (at a membrane holding voltage of -70 mV). Hexamethonium (100 microM) applied only to the ganglion abolished both electrically- and distension-evoked excitatory postsynaptic currents, suggesting activation of nicotinic acetylcholine receptors. The decay time-course of distension-evoked single excitatory postsynaptic currents was characterized by one, or, less commonly, by two exponentials. The decay time-constant histograms of distension-evoked single excitatory postsynaptic currents exhibited main kinetic components of 8.1 +/- 2.3 ms and 8.2 +/- 2.5 ms (peak +/- S.D.) at -70 and -110 mV membrane holding voltages, respectively. Longer time-constants ranging up to 51 ms were also observed. The number of the distension-evoked excitatory postsynaptic currents with a decay time-constant higher than 20 ms, as well as their mean amplitude, were significantly lower at -110 mV than at -70 mV membrane potential levels, in contrast to the currents with a decay time-constant lower than or equal to 20 ms. The results suggest that colonic afferent nerve fibres activate in the mouse superior mesenteric ganglion neurons a few populations of the postsynaptic nicotinic acetylcholine receptors with different channel kinetics, which are characterized by a lack of voltage sensitivity within -70 to -110 mV membrane potential range, except those with comparatively slow channel kinetics, which are possibly blocked by membrane hyperpolarization. PMID- 9740410 TI - Analysis and biophysical interpretation of bistable behaviour in thalamocortical neurons. AB - Thalamocortical neurons display a wide spectrum of activity patterns that are the expressions of the non-linear interactions between the various voltage-gated ion channels. Here, we show how bistable behaviour can emerge in these neurons, and how it is brought about by the steady-state residual ("window") component of IT, the low-threshold Ca2+ current. In particular, we present results that describe the dependence of bistability on two system parameters: the injected current and the leakage conductance. In addition, we provide a biophysical interpretation of these results by means of the properties of the electrical circuit representing the neuron membrane. PMID- 9740411 TI - Measuring time to pregnancy. Methodological issues to consider. PMID- 9740412 TI - The use of fecundability in epidemiological surveys. PMID- 9740413 TI - Vasectomy related infertility: a major and costly medical problem. AB - Of a group of 860 men who attended a private infertility clinic in Western Australia, 80 (9.3%) presented with vasectomy-related infertility. Of these men, 73 (91%) requested treatment due to re-marriage. The median age of the men was 42.5 years and their present partners were approximately 10 years younger. The median vasectomy interval in the men in this study was 9 years. Treatment of vasectomy-related infertility included vasectomy reversal procedures, donor insemination and both in-vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). It is suggested that care must be taken in counselling such patients prior to a vasectomy. The cryopreservation of semen prior to vasectomy would also make much of this type of infertility treatment unnecessary. PMID- 9740414 TI - Vasectomy: an effective form of contraception. PMID- 9740415 TI - A French survey on gynaecological laparoscopy. PMID- 9740416 TI - Transvaginal access heralds the end of standard diagnostic laparoscopy in infertility. PMID- 9740417 TI - Current status of the molecular diagnosis of Y-chromosomal microdeletions in the work-up of male infertility. Initiative for international quality control. PMID- 9740418 TI - Principles of sequence-tagged site selection in screening for Y deletions. PMID- 9740419 TI - Should the use of assisted reproduction techniques be deregulated? The UK experience: options for change. PMID- 9740420 TI - Creutzfeldt-Jakob disease (CJD) and assisted reproductive technology (ART). Quantification of risks as part of informed consent. PMID- 9740421 TI - Are we on the verge of a new era in ART? PMID- 9740422 TI - Venous thromboembolism and the pill. Learning from the past: a response from The Committee on Safety of Medicines. PMID- 9740423 TI - Outcome from consecutive in-vitro fertilization/intracytoplasmic sperm injection attempts in the final group treated with urinary gonadotrophins and the first group treated with recombinant follicle stimulating hormone. AB - In the absence of specific dose equivalency data, the aim of this study was to compare the clinical results during the cross-over from menopausal urinary products (human menopausal gonadotrophin; HMG) to recombinant follicle stimulating hormone (FSH) follitrophin beta (FSHr) in order to determine whether the manufacturer's recommendation for equivalence of ampoule to ampoule (50 IU FSHr:75 IU HMG) would prove clinically correct. A total of 353 consecutive in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) treatment cycles was studied between 1st September 1996 and mid-February 1997. This included cycles in the last 191 women receiving HMG and the first 162 taking FSHr. All were down-regulated using a gonadotrophin releasing hormone (GnRH) agonist long protocol method from day 1 of the cycle. Greater efficacy was seen in the HMG group in terms of days of stimulation required, need to increase dosage, cycle discontinuation, number of follicles punctured, the numbers of oocytes retrieved and their quality. The hormonal response to stimulation assessed by oestradiol concentrations on days 5, 8 and day of human chorionic gonadotrophin (HCG) was significantly lower in the FSHr group. The ratio of oestradiol per follicle and per oocyte was significantly lower in the FSHr group. There was a highly significant increase in cost with FSHr therapy. Clinical pregnancy rates were 14% per cycle with FSHr and 20% per cycle with HMG. PMID- 9740424 TI - Effects of profound suppression of luteinizing hormone during ovarian stimulation on follicular activity, oocyte and embryo function in cycles stimulated with purified follicle stimulating hormone. AB - The effects of profound suppression of circulating luteinizing hormone (LH) during the follicular phase of in-vitro fertilization cycles were explored in normal women during treatment with a gonadotrophin-releasing hormone analogue and exogenous purified follicle stimulating hormone. Ovarian responses to treatment and the capacity of supernumerary embryos to undergo blastocyst formation were examined in groups of patients defined by the concentration of plasma LH in the mid-follicular phase. Concentrations < or = 0.5 IU/I diagnosed the group with profoundly suppressed LH (30. Complication rate was 5.1% of pinpoint perforations of the tube. With Cox's statistical model, none of the parameters analysed was statistically predictive of intrauterine pregnancy. We conclude that the greater accuracy of diagnosis by falloposcopy may indicate that it should be incorporated into the initial screening of infertile patients. PMID- 9740432 TI - Oocyte donation in Israel: a study of 1001 initiated treatment cycles. AB - There are numerous studies concerning pregnancy rates in oocyte donation, yet only a handful report the obstetric outcome in such pregnancies. The purpose of this study was to assess factors that influence pregnancy rates, to determine the incidence of complications, and to evaluate obstetric outcome in pregnancies resulting from oocyte donation. This study included 423 oocyte recipients who underwent 1001 oocyte donation cycles at the Oocyte Donation Programme, In-Vitro Fertilization (IVF)-Embryo Transfer Unit, Herzlia Medical Center, Israel. Donors were all healthy women < 34 years old who underwent IVF themselves. In 873 cycles, fertilization occurred and embryo transfer was performed, resulting in 194 clinical pregnancies. Pregnancy rates (PR) significantly declined with the increase in number of previous attempts, and with increasing age of recipient (36.8%/embryo transfer in patients < or = 30 compared to 17.8% in patients > 40 years old). A significant increment in PR was noted with the increasing number of embryos transferred. The overall PR was 22.2%/embryo transfer. However, in young amenorrhoeic patients with normal karyotypes undergoing their first cycle, PR was 52.2%; the 'take home baby' rate was 38.3% per patient undergoing embryo transfer and 17.8% per embryo transfer cycle. A significant increase in the incidence of pregnancy-induced hypertension and a higher proportion of abortions were noted in older patients. A significantly higher incidence of prematurity and low birthweight was observed in multiple pregnancies. PMID- 9740433 TI - The contribution of subtle oocyte or sperm dysfunction affecting fertilization in endometriosis-associated or unexplained infertility: a controlled comparison with tubal infertility and use of donor spermatozoa. AB - This study aims to determine the relative contribution of oocyte and/or sperm dysfunction to the reduction of fertilization rates in vitro in cases of minor endometriosis and prolonged unexplained infertility. The results of in-vitro fertilization (IVF) treatment with ovarian stimulation have been compared between couples with the above conditions and women with tubal infertility (as control for oocyte function) and the use of donor spermatozoa (as control for sperm function). Fertilization and cleavage rates using husband's spermatozoa were significantly reduced in endometriosis couples (56%, n = 194, P < 0.001) and further significantly reduced in couples with unexplained infertility (52%, n = 327, P < 0.001) compared with tubal infertility (60%, n = 509). Using donor spermatozoa the rates were the same as using husband's spermatozoa in tubal infertility (61%, n = 27) or endometriosis (55%, n = 21) but significantly though only partly improved with unexplained infertility (57%, n = 60, P < 0.02). In unexplained infertility, a significantly increased proportion of couples experienced complete failure of fertilization and cleavage in a cycle (5-6% versus 2-3%). However, complete failure was not usually repetitive, and the affected couples did not account for the overall reduction in fertilization and cleavage rates, which remained significantly lower in the rest of the unexplained and endometriosis groups. Implantation and pregnancy rates appeared similar in all groups. The benefit of IVF treatment in cases of minor endometriosis and prolonged unexplained infertility is due to superabundance of oocytes obtained by stimulation. The reduction in natural fertility associated with endometriosis appears to be at least partly due to a reduced fertilizing ability of the oocyte. In unexplained infertility, there is distinct impairment due to otherwise unsuspected sperm dysfunction but probably also oocyte dysfunction. PMID- 9740434 TI - Patient satisfaction with the management of infertility. AB - The objective of this study was to assess patient satisfaction with the investigation and initial management of infertility. A postal questionnaire survey was carried out of 1366 women attending outpatient clinics for the investigation and initial management of infertility at 12 hospitals throughout Scotland. The response rate to the questionnaire was 59% (806/1366). Overall, 87% of responders were satisfied or very satisfied with their care but a number of deficiencies were identified. Thirty-nine per cent had never been asked to bring their partner to the clinic and 86% felt they had not been given enough help with the emotional aspects of infertility. Forty-seven per cent felt they were not given a clear plan for the future and 23% of those who had been given drug treatments reported receiving little or no information about the treatment or possible side-effects. Overall, only a third had been given any written information and 78% expressed a wish for more written information. Women ranked 'the information and explanation given' and the 'attitude of the doctor at the clinic' highly in comparison to other aspects of their care, including 'help with the emotional aspects of infertility'. In general women were satisfied with their care but improvements may be made by giving more explanation and written information and by adopting a more couple-centred approach. Where resources allow, clinics should take steps to address the emotional aspects of infertility. PMID- 9740435 TI - Malignancy may adversely influence the quality and behaviour of oocytes. AB - A case series of eight cycles of in-vitro fertilization (IVF) in five women diagnosed with malignant disorders is presented. These patients chose to defer definitive treatment for a chance for preservation of potential fertility. The response of these patients to ovarian stimulation, and the outcome, was compared with 17 IVF cycles in 12 age-matched patients with isolated tubal infertility. An apparent adverse influence of malignant disease on the quality and behaviour of oocytes was observed. Despite a comparable total number of oocytes per cycle in the two groups, a significantly reduced percentage of mature oocytes was retrieved per cycle from patients with malignant diseases. The oocytes from patients with malignant disorders were of a poorer quality and exhibited a significantly impaired fertilization rate compared to the controls. We propose that neoplastic processes, irrespective of the site or cell of origin, may have a detrimental impact on the biology of oocytes, an effect akin to that seen on spermatozoa in men with certain malignancies. PMID- 9740436 TI - The effects of a copper-intrauterine device on the uterine artery blood flow in regularly menstruating women. AB - The aim of the study was to evaluate the effect of a copper-intrauterine device (IUD) on uterine artery blood flow during the midluteal phase and on the first day of the menstrual cycle using pulsed colour Doppler ultrasonography. Twenty one regularly menstruating women (18-45 years) who were willing to use copper-IUD contraception participated in the study. The patients were first examined without the IUD in the midluteal phase 6-9 days before the expected onset of menstruation and on the first day of menstruation, after which the IUD was inserted. Three months later the patients were examined again on the corresponding cycle days. The patients estimated the level of dysmenorrhoeic pain with a scoring system. Transvaginal ultrasonography with colour flow imaging was used to measure the pulsatility index (PI) in the uterine arteries. There were no significant changes in the uterine artery blood flow after the insertion of the IUD during menstruation or in the midluteal phase. In patients with increased IUD-related pain during menstruation (n = 5), however, there was a decrease in PI (2.87 +/- 0.52 versus 2.41 +/- 0.23, P = 0.05) after IUD insertion. The decrease in the mean PI was present in all five patients. In conclusion, copper-IUD does not induce any major changes in the resistance of the uterine artery blood flow, although during menstruation in patients with increased menstrual pain after IUD insertion there seems to be a decrease in the uterine artery PI. PMID- 9740437 TI - The ultrastructure of human endometrium is altered by administration of intrauterine levonorgestrel. AB - We studied the effect of intrauterine administration of levonorgestrel (LNG) on the ultrastructure of the endometrium. Twenty-one endometrial biopsy specimens, collected from nine fertile women during normal menstrual cycles and after 1, 3 or 6 months of use of a levonorgestrel-releasing intrauterine contraceptive system (LNG IUS), were studied using transmission and scanning electron microscopy. During the 6 month exposure to LNG IUS, changes took place in the endometrium. The glandular epithelial cells became lower. The junctional complexes between epithelial cells remained unchanged, whereas the lateral microvillar interdigitations became more prominent. The basal lamina under the epithelium became wavy but remained uniform and practically uninterrupted; only solitary epithelial cell protrusions through the basal lamina were seen. The stromal cells were largely decidualized. We conclude that in parallel with the generally known cellular effects, the use of the LNG IUS results in distinct changes in the basal lamina between the endometrial epithelial and stromal cells. The especially well-developed and uninterrupted basal lamina may be involved in the mechanism of the LNG IUS-induced endometrial suppression. Furthermore, the complex intercellular junctions between the epithelial cells, normally loosening around the time of implantation, persist during the local administration of levonorgestrel. This may have a pivotal role in the contraceptive effect of the LNG IUS. PMID- 9740438 TI - The use of intracytoplasmic sperm injection with electroejaculates from anejaculatory men. AB - Electroejaculation has been successfully used for sperm procurement in anejaculatory men desiring fertility. However, electroejaculates typically have normal sperm numbers but poor motility, morphology, and functional deficiencies. Here we report the pregnancy outcome of a series of couples undergoing combined electroejaculation and in-vitro fertilization (IVF) with intracytoplasmic sperm injection (ICSI). In all, 13 couples underwent a total of 18 cycles. The aetiologies of anejaculation included history of retroperitoneal lymph node dissection for testicular cancers, spinal cord injury and psychogenic causes. ICSI was performed on 192 oocytes, resulting in a fertilization rate of 75.5%. A total of 15 embryo transfers were performed using a total of 51 embryos. Clinical pregnancy rate, as defined by positive fetal heart rate(s) using vaginal sonography, was 55.6% per retrieval; implantation rate was 33.3% per embryo. These rates appear to be similar to those obtained in standard IVF for non-male factor infertility, or ICSI for male factor infertility. The use of ICSI for electroejaculates undoubtedly provides these couples with the highest chance of pregnancy. PMID- 9740439 TI - Semen preparation by standard swim-up versus swim-up with test yolk buffer incubation in intrauterine insemination: a randomized study. AB - In order to compare the standard swim-up semen preparation with and without test yolk buffer (TYB) incubation in intrauterine insemination (IUI), we conducted a prospective multicentre randomized trial. A total of 121 infertile couples with male factor (n = 52) or unexplained infertility (n = 69) was randomly assigned to two groups following ovulation induction. Semen was prepared by standard swim-up in group A (n = 64) and by swim-up followed by TYB incubation in group B (n = 57). A maximum of two IUI cycles was performed. A total of 104 cycles was performed in the swim-up group and 90 in the TYB group. Overall, 15 pregnancies were achieved in group A and 23 in group B, with an overall pregnancy rate of 24.8 and 50.0% per patient respectively (chi2(1), P < 0.05). In the male factor group, pregnancy was achieved in six out of 24 couples (25%) following standard swim-up and in six out of 28 (21.4%) following swim-up and TYB incubation (chi2(1), not significant). In the unexplained infertility group, pregnancy was recorded in nine out of 40 couples (22.5%) following standard swim-up and in 17 out of 29 couples (58.6%) following swim-up and TYB incubation (chi2(1), P < 0.05). PMID- 9740440 TI - Oxidative damage to DNA in human spermatozoa does not preclude pronucleus formation at intracytoplasmic sperm injection. AB - We present the first evidence that genetically damaged human spermatozoa are able to form normal pronuclei in oocytes after intracytoplasmic sperm injection (ICSI). The role of reactive oxygen species (ROS) as a cause of chromatin and DNA damage is well recognized. The same class of molecule can be found in the semen of males with severe infertility, who remained infertile until the advent of ICSI. In this study we have investigated the role of ROS in the induction of chromatin damage, DNA strand breakage and the subsequent ability of spermatozoa to decondense and form pronuclei after ICSI. Spermatozoa from normozoospermic men participating in our research programme were exposed to oxidizing environments created by co-incubation with hydrogen peroxide, reduced nicotinamide adenine dinucleotide phosphate (NADPH) or activated white cells. The subsequent ability of the spermatozoa to decondense in vitro was examined using sequential incubations in EDTA, dithiothreitol and sodium dodecyl sulphate, and the amounts of DNA strand breakage were assessed using an in-situ nick translation protocol. Finally, cells exposed to hydrogen peroxide, NADPH and activated leukocytes were microinjected into hamster oocytes, and their ability to decondense and form normal pronuclei was determined. The results indicate that human sperm chromatin becomes cross-linked under conditions of oxidative stress and exhibits increased DNA strand breakage, yet the rate of pronucleus formation is no different from that of untreated control cells. The ability of genetically damaged spermatozoa to achieve normal fertilization following ICSI has implications for the practice of this form of assisted conception therapy. PMID- 9740441 TI - The outcome of intracytoplasmic injection of fresh and cryopreserved epididymal spermatozoa from patients with obstructive azoospermia--a comparative study. AB - The aim of our study was to compare the outcome of intracytoplasmic sperm injection (ICSI) with fresh and frozen-thawed epididymal spermatozoa retrieved by percutaneous epididymal sperm aspiration (PESA) or microepididymal sperm aspiration (MESA) from patients with obstructive azoospermia. A retrospective analysis of consecutive ICSI cycles was performed, comparing the outcome in 24 patients with obstructive azoospermia undergoing surgical sperm aspiration by MESA (7 cycles) or PESA (17 cycles). In 23 of 24 patients, excess spermatozoa were cryopreserved. Following thawing, 21 ICSI cycles were performed (11 cycles after MESA, 10 after PESA). No statistically significant differences were noted in all parameters examined in ICSI cycles with fresh or cryopreserved spermatozoa from the same patients. Comparing all ICSI cycles with fresh and frozen-thawed epididymal spermatozoa, the rates of two-pronuclear fertilization (56% versus 53%), embryo cleavage (90% versus 86%), implantation (10% versus 14%), clinical pregnancy per embryo transfer (32% versus 37%) and delivery/ongoing pregnancy rate (27% versus 26%) were not statistically different. The cumulative ongoing pregnancy rate per sperm retrieval procedure was 46%, respectively. We conclude that the clinical outcome of ICSI with fresh and frozen-thawed spermatozoa after retrieval by PESA was similar to that by MESA. Epididymal sperm cryopreservation in patients with obstructive azoospermia is feasible and efficient using a simple freezing protocol and should be offered to optimize the yield of pregnancies achieved following such procedures. PMID- 9740442 TI - Effects of co-trimoxazole, erythromycin, amoxycillin, tetracycline and chloroquine on sperm function in vitro. AB - This in-vitro study was designed to investigate the effects of commonly prescribed antibiotics on sperm movement characteristics, viability and the ability of spermatozoa to undergo the acrosome reaction. Spermatozoa were obtained by swim-up from normozoospermic semen and cultured for 24 h with increasing concentrations of co-trimoxazole, erythromycin, amoxycillin, tetracycline and chloroquine. Tetracycline at concentrations as low as 2.5 microg/ml led to a significant dose-dependent inhibition in percent rapid-moving spermatozoa, mean path velocity (VAP), straight-line velocity (VSL) and curvilinear velocity (VCL), but at 50 microg/ml tetracycline all spermatozoa were static. Erythromycin had significant effects on rapid movement, VAP, VSL and VCL only at concentrations >100 microg/ml. In contrast, percent rapid-moving spermatozoa was significantly enhanced at low concentrations of chloroquine (5 microg/ml), but significantly inhibited by higher concentrations. Co-trimoxazole did not adversely affect percent rapid-moving spermatozoa below 500 microg/ml, at which concentration movement was decreased by 34%. The mean lateral head displacement (ALH) was significantly enhanced by 5 microg/ml co-trimoxazole and reduced at 1 mg/ml erythromycin. The effects of these drugs were mostly irreversible. Amoxycillin had no effect on sperm movement characteristics over the dose range used, though it inhibited viability at high doses. Viability was significantly reduced at concentrations of all drugs which affect rapid sperm movement; these concentrations of drugs did not appear to affect the ability of spermatozoa to undergo the acrosome reaction. The results from this study, when combined with known effects on spermatogenesis, should facilitate the choice of drugs for the treatment of both genitourinary and unrelated infections in men who are attempting to conceive. PMID- 9740443 TI - A birth in non-mosaic Klinefelter's syndrome after testicular fine needle aspiration, intracytoplasmic sperm injection and preimplantation genetic diagnosis. AB - Non-mosaic Klinefelter patients are generally azoospermic due to primary testicular failure. Nevertheless, in some cases, testicular spermatozoa may be recovered and utilized to fertilize oocytes via intracytoplasmic sperm injection (ICSI). As the risk for an increased number of gonosomes in these spermatozoa is unclear, preimplantation genetic diagnosis (PGD) may be attempted in the resulting embryos. In the present study, we report our experience with the combined approach of sperm retrieval by testicular fine needle aspiration (FNA), ICSI and PGD in seven consecutive non-mosaic Klinefelter individuals. In four patients, between one and five spermatozoa were retrieved in five out of nine consecutive attempts. In a fifth patient, only 10 round spermatids could be isolated. Mature spermatozoa were injected into a total of 16 metaphase-II oocytes, of which 11 (69%) remained intact. Two distinct pronuclei (2PN) were observed in four oocytes (36%) while a single pronucleus (1PN) was documented in two oocytes. Five cleavage stage embryos developed from the oocytes of two couples. Upon the request of one couple, their three embryos (two derived from 1PN oocytes) were transferred without PGD but pregnancy was not achieved. PGD by fluorescence in-situ hybridization (FISH) was performed in the two embryos of the other couple which were derived from normal fertilization. PGD results of one embryo were 18,18,X,X,Y, the embryo was not transferred and FISH analysis of the remaining blastomeres identified variable chromosome numbers in the nuclei. The second embryo was diagnosed as normal and was transferred, resulting in a successful pregnancy and birth. In conclusion, the results of this report indicate that a pregnancy and birth may be attained in azoospermic non-mosaic Klinefelter individuals by testicular FNA combined with ICSI. Due to the unknown risk of gonosomes aneuploidy in embryos from Klinefelter patients, PGD or prenatal diagnosis should be recommended. PMID- 9740444 TI - Fertilization, pregnancy and embryo implantation rates after ICSI with fresh or frozen-thawed testicular spermatozoa. AB - This study aimed to determine whether fertilization and implantation rates after intracytoplasmic sperm injection (ICSI) with fresh or frozen-thawed testicular spermatozoa were comparable. Between 1 January 1996 and 31 December 1996, 65 ICSI cycles with testicular spermatozoa and 35 cycles with frozen-thawed testicular spermatozoa were carried out. In 50 out of 65 ICSI cycles, testicular spermatozoa could be retrieved and in 34 out of 35 cycles carried out with frozen-thawed testicular spermatozoa, motile spermatozoa could be recovered. The fertilization rate after ICSI with frozen-thawed testicular spermatozoa was significantly lower (71.1%; P < or = 0.008) than with fresh testicular spermatozoa (79.3%). The pregnancy rate was similar for both groups (38.2 and 26.5 %). The implantation rate per transferred embryo, however, was significantly lower in the frozen thawed rather than in the fresh testicular sperm group (9.1 versus 24.6%; P = 0.001). The live birth rate per transferred embryo was also higher in the group in which fresh testicular spermatozoa were used (18.8 versus 7.9% P = 0.043). This retrospective study shows that is possible to achieve a high fertilization rate after ICSI with both fresh and frozen-thawed testicular spermatozoa but implantation and live birth rates per transferred embryo, however, are significantly lower after ICSI with frozen-thawed than with fresh testicular spermatozoa. PMID- 9740445 TI - Successful pregnancy after spermatid injection. AB - We present nine cases of spermatid intracytoplasmic injection for the treatment of non-obstructive azoospermia. In eight cases, no elongated spermatids or spermatozoa were found in previous spermiograms or testicular biopsies. In these patients, treatment was performed using ejaculated (n = 6) and testicular (n = 2) retrieved round spermatids (Sa type). In cases where ejaculated round spermatids were used, they were isolated on the day before oocyte retrieval and left in culture for 24 h before intracytoplasmic sperm injection (ICSI). No pregnancy was obtained in either group, although culturing seemed to increase the fertilization rate. In one other case, elongated spermatids were observed in the previous spermiogram and thus a normal ICSI procedure was scheduled. However, on the day of oocyte retrieval, no spermatids could be recovered from fresh sequential ejaculates, and a testicular open biopsy was then performed. Both round and elongated spermatids were found in the testicular tissue, but only the more mature germinal cells (Sb2) were injected. From this case, a normal pregnancy was obtained which resulted in the birth by Caesarean section at 37 weeks of gestation of a normal healthy baby girl, weighing 2700 g. PMID- 9740447 TI - Differential effect of exogenous human chorionic gonadotrophin on progesterone production from normal or malfunctioning corpus luteum. AB - To examine whether luteal phase defect is, in part, causally related to insufficient gonadotrophin stimulation, we compared the relation of the increment of serum progesterone concentrations in response to human chorionic gonadotrophin (HCG) with its basal level at mid-luteal phase. Thirty-eight naturally cycling infertile women aged between 27-41 years old were evaluated for hormonal responses to HCG injection at the mid-luteal phase. We measured luteinizing hormone (LH), follicle stimulating hormone (FSH), oestradiol and progesterone concentrations, before and 1, 2 and 3 h after the administration of HCG (5000 IU, i.m.) 7 days after ovulation verified by ultrasonography. Eleven out of 38 women exhibited progesterone concentrations below 10 ng/ml (low progesterone group), and those remaining showed progesterone concentrations of > or = 10 ng/ml (normal progesterone group). The basal LH, FSH and oestradiol concentrations were essentially the same in both groups. Progesterone concentrations rose significantly 1 h after the injection and levelled off thereafter. The increment of progesterone concentrations at 1 h in the normal progesterone group was 5.7 ng/ml on the average, whereas that in low progesterone group was 1.1 ng/ml. Furthermore, the percentage increase in progesterone concentrations at 1 h in the normal progesterone group was significantly greater than that in the low progesterone group. Both groups equally exhibited significant but marginal increases in oestradiol concentrations 1 h after the injection. LH and FSH concentrations at 3 h decreased significantly in both groups. In summary, HCG readily stimulates progesterone production in normally functioning corpus luteum whereas its stimulatory effect is minimal on malfunctioning corpus luteum. This suggests that luteal phase defect is not caused by inadequate gonadotrophin stimulation and, therefore, does not benefit from HCG administration. PMID- 9740446 TI - The relationship between follicular fluid aspirate volume and oocyte maturity in in-vitro fertilization cycles. AB - As a consequence of multiple follicular growth during ovarian stimulation for in vitro fertilization (IVF), follicles of varying sizes often yield oocytes that vary in maturity and morphology of the oocyte-cumulus-corona complex. The objective of this prospective study was to explore the relationship between follicular fluid aspirate volume and the oocyte's developmental potential in an IVF treatment cycle. In total 9933 follicles were studied from 400 patients who underwent 535 consecutive IVF treatment cycles at St James's University Hospital, Leeds, UK, between February 1995 and February 1996. The volume of each individual follicle aspirated was recorded and related to the probability of obtaining an oocyte, its fertilizing capacity, the cleavage rate and the quality of embryos derived. We found no statistically significant difference in oocyte recovery rates between follicles with an aspirate volume < or = 1 ml and follicles with a volume > 1 ml. Although oocytes obtained from follicles with an aspirate volume > or = 1 ml showed a significantly lower fertilization rate, they went on to cleave at the same rate as oocytes obtained from larger follicles and resulted in embryos of comparable quality. Furthermore, there was no statistically significant difference in the implantation, clinical pregnancy or live birth rates per cycle between embryos derived from follicles with an aspirate volume < or = 1 ml and those derived from follicles with an aspirate volume > 1 ml. We conclude that follicular size and the oocyte's developmental potential in the stimulated ovary are not closely related and can be independent. This is in contrast to the Graafian follicle and the pre-ovulatory oocyte in the natural cycle. PMID- 9740448 TI - Comparison of fertilization, cleavage and pregnancy rates of oocytes from large and small follicles. AB - Ovarian stimulation for in-vitro fertilization (IVF) causes development of several cohorts of follicles. At the time of oocyte collection, oocytes are thus retrieved from a wide range of follicles of different sizes and developmental stages. A relationship between size of follicles and pregnancy rates has earlier been demonstrated. The aim of the present study was to compare fertilization, cleavage and pregnancy rates between oocytes retrieved from large and small follicles in conventional IVF and intracytoplasmic sperm injection (ICSI). A total of 200 conventional IVF patients and 175 ICSI patients underwent oocyte retrieval where oocytes from both large and small follicles were collected. A follicle with a volume of > or = 2 ml, corresponding to a follicular diameter > or = 16 mm as determined by ultrasound, was regarded as a large follicle. Only one cycle from each patient was included. Fertilization and cleavage rates were calculated per patient for oocytes from large and small follicles. The mean fertilization and cleavage rates for conventional IVF and ICSI cycles were calculated. Comparison of pregnancy rates was performed for patients receiving embryos derived from oocytes of only large or only small follicles. For conventional IVF patients, fertilization rates were 71.4 and 58.1% (P < 0.01, Wilcoxon paired test) for oocytes of large and small follicles respectively. The corresponding cleavage rates were 95.4 and 93.9% respectively. The pregnancy rate for the two groups was 47% (60/127) and 15% (2/13) (P < 0.05, chi2 test). For ICSI patients the fertilization rate was 72.0 and 71.1% for oocytes of large and small follicles respectively. The corresponding cleavage rate was 93.0 and 91.1%. The pregnancy rate in the two groups was 41% (46/113) and 42% (5/12). The results show that oocytes from smaller follicles also yield fertilization and pregnancies, although in conventional IVF to a lesser extent than oocytes from larger follicles. For IVF cycles, a higher proportion of immature oocytes (which are normally not included in the ICSI procedure) in the group of oocytes from small follicles is most probably the explanation for the lower fertilization rate. The decrease in pregnancy rate with oocytes from small follicles in the IVF cycles was not observed in the ICSI cycles. The possibility of evaluating the degree of oocyte maturation prior to fertilization may be an advantage of the ICSI technique. This suggests that the disadvantages of oocytes from small follicles might be overcome by means of ICSI. PMID- 9740449 TI - Development of hatching blastocysts from immature human oocytes following in vitro maturation and fertilization using a co-culture system. AB - Recently, in-vitro maturation (IVM) of immature human oocytes recovered from non stimulated follicles has been applied in the treatment of infertility. However, in previous reports, very few embryos cultured in conventional medium have reached the expanded blastocyst stage following in-vitro maturation and fertilization (IVM/IVF). The objective of this study was to investigate whether the developmental competence of human embryos following IVM/IVF could be enhanced by the use of a human ampullary cell co-culture system. Immature human oocytes were aspirated from small follicles at Caesarean section and then cultured in medium containing human menopausal gonadotrophin for 36 to 48 h, followed by insemination. Zygotes were randomly cultured either in conventional culture medium alone or in the co-culture system. Of 48 embryos cultured in conventional medium alone, all arrested at the 2-16-cell stage on day 3 after insemination. Of 46 embryos cultured in the co-culture system, 26 embryos (56.5%) arrested at the 2-16-cell stage. Six embryos (13%) developed to the morula stage. Fourteen embryos (30.4%) developed to expanded blastocysts and two blastocysts were hatching on day 7 after insemination. We conclude that co-culture significantly enhances the development of blastocysts in embryos resulting from IVM/IVF. PMID- 9740450 TI - A modified method of intracytoplasmic sperm injection without the use of polyvinylpyrrolidone. AB - In a controlled study we compared the outcome of intracytoplasmic sperm injection (ICSI) performed by two different methods. The oocytes from 20 patients were equally divided into two groups and injected either by conventional ICSI using polyvinylpyrrolidone (PVP) or by a modified PVP-free ICSI procedure. While in the conventional ICSI method the spermatozoon is aspirated into the injection pipette, in the modified ICSI procedure the spermatozoon is attached to the end of the narrow micropipette by aspirating its tail. The sperm head is never drawn into the pipette. Accordingly, even a fast-moving spermatozoon can be 'caught' easily. As a result of such an aspiration the spermatozoon loses its motility. Therefore, PVP is required neither to slow down the movement of the spermatozoon nor to facilitate the movement of the solution in the injection pipette. A total of 230 mature oocytes were injected by both methods and the results were analysed. No differences were observed in survival rate between the two ICSI procedures (89% and 91%, respectively). However, the proportion of normally fertilized oocytes was significantly higher after microfertilization by modified ICSI (74%) when compared with the outcome of the conventional ICSI method (62%). The frequency of abnormal fertilization was not influenced by the method of ICSI used. The cleavage rate and quality of resulting embryos were also comparable. In conclusion, we have demonstrated a modified ICSI method which does not require the use of PVP. When compared with the conventional ICSI procedure, even better fertilization rates can be achieved. The proposed ICSI modification may provide an alternative procedure for elimination of the potentially harmful effects which may be associated with conventional ICSI. PMID- 9740451 TI - Relationship between human in-vitro fertilization and intracytoplasmic sperm injection and the zona-free hamster egg penetration test. AB - The zona-free hamster egg penetration test (HEPT) is widely used for evaluating the fertilizing ability of human spermatozoa. However, the relationship between the HEPT and microassisted fertilization has yet to be determined. To evaluate the efficiency of HEPT in selecting the most appropriate method of in-vitro fertilization (IVF), including intracytoplasmic sperm injection (ICSI) in couples with male factor infertility, clinical laboratory data was analysed retrospectively. The patients were divided into groups according to the sperm penetration index as determined by the HEPT: group A (sperm penetration index = 0), group B (sperm penetration index < 15) and group C (sperm penetration index > or = 15). A total of 405 oocytes were collected and inseminated by conventional methods in 69 couples with male factor infertility. In all, 31 out of 148 (20.9%) oocytes fertilized in group A; 35 out of 117 (29.9%) in group B; and 73 of 140 (52.1%) in group C. The clinical pregnancy rates per transfer in groups A, B and C were 0% (0/13), 0% (0/14) and 25.9% (7/27) respectively. Both the fertilization rate and pregnancy rate in group C was significantly higher than in groups A and B. ICSI was carried out in a total of 57 couples and 334 oocytes in metaphase II stage were manipulated. The normal fertilization (2 pronuclear) rate per oocyte was 65.6 +/- 26.0% (mean +/- SD). Out of 127 oocytes, 76 (59.8%) fertilized in group A, 57 out of 87 oocytes (65.5%) in group B and 86 out of 120 oocytes (71.7%) in group C. Of the 56 transfers, 17 clinical pregnancies were obtained, giving an average pregnancy rate of 30.4% per transfer. The clinical pregnancy rates per transfer in groups A, B and C were 17.4% (4/23), 40.0% (4/10) and 39.1% (9/23) respectively. No significant differences were observed in the fertilization rates or in the pregnancy rates between the three groups. In addition, there were no differences in the fertilization and pregnancy rates between the ICSI and IVF patients in group C. These findings suggest that the results of the HEPT are well correlated with the fertilizing ability of human spermatozoa in the patients treated by conventional IVF. Couples suffering from male factor infertility with a sperm penetration index of < 15 (as determined by HEPT) should consider treatment with ICSI, while those with a sperm penetration index of > or = 15 should attempt conventional IVF. PMID- 9740452 TI - Use of a plant enzyme preparation (Coronase) instead of hyaluronidase for cumulus cell removal before intracytoplasmic sperm injection. AB - The aim of this study was to compare the efficiency of a plant enzyme preparation (Coronase) with animal extracted hyaluronidase to remove cumulus cells before intracytoplasmic sperm injection (ICSI). The first part of the study was performed on mouse oocytes and embryos. Coronase displayed a similar efficiency to that of hyaluronidase for removing cumulus cells and the same percentage of activated oocytes was obtained with both techniques. However, prolonged incubation in Coronase, 120 min, led to a degeneration of oocytes. Incubation of 2-cell mouse embryos for 10 min with Coronase did not affect their subsequent in vitro development to blastocyst. Coronase was then compared to hyaluronidase in the treatment of human oocytes prior to ICSI. The time required for total denudation was slightly longer using Coronase (98 s +/- 25 s versus 84 s +/- 24 s respectively for Coronase and hyaluronidase; P < 0.01). However, the two pronuclear (2PN) fertilization rate (70/103 versus 63/107 respectively for Coronase and hyaluronidase, not significant) and the percentage of embryos with a good morphology (39/74 versus 32/67 respectively for Coronase and hyaluronidase, not significant) were identical with both treatments. In conclusion, Coronase displays an efficiency close to that of hyaluronidase, without any adverse effect on oocytes, and may be preferable for human use. PMID- 9740454 TI - Uterine leiomyomata--a feature of acromegaly. AB - In order to assess the prevalence of leiomyomata in patients diagnosed with acromegaly, files of all women so diagnosed were obtained (n = 25). Eight of these patients had undergone hysterectomy. In eight of the remaining patients, assessment of the uterus was performed by gynaecological examination and transvaginal ultrasound. The prevalence of leiomyomata was 81% (13/16) in patients who had undergone hysterectomy (8/8) or who underwent gynaecological examination (5/8). In conclusion, the very high prevalence of leiomyomata in patients diagnosed with acromegaly warrants the inclusion of growth hormone excess as a cause of leiomyomata. Leiomyomata are a feature of the organomegalic syndrome associated with acromegaly. PMID- 9740453 TI - Attitude of potential users in Sicily towards preimplantation genetic diagnosis for beta-thalassaemia and aneuploidies. AB - This study aims to report the willingness of different populations of high-risk couples to undergo preimplantation genetic diagnosis (PGD) for beta-thalassaemia as an alternative to prenatal genetic diagnosis (PND), and the willingness of infertile couples to undergo PGD for aneuploidies. An information sheet and questionnaire presenting PGD and PND procedures were distributed to four population types: 54 high-risk couples for beta-thalassaemia coming for their first PND (population A); 51 similar couples coming for their second or further PND without previous experience of therapeutic abortion (population B-na); 50 similar couples coming for their second or further PND with previous experience of therapeutic abortion for beta-thalassaemia-affected fetus (population B-ab); and 74 infertile couples undergoing routine in-vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) (population C). Favourable first impressions towards PGD compared with PND were observed in all four populations in the following proportions: 79.6% population A; 76.5% population B-na; 92.0% population B-ab; and 96.0% population C. Willingness to undergo PGD for beta thalassaemia was as follows: 44.4% population A; 47.1% population B-na; and 72.0% population B-ab. We conclude that previous experience of PND for beta thalassaemia is a crucial point in the willingness to accept the PGD procedure, and that couples belonging to population B-ab are the most suitable to undergo PGD for beta-thalassaemia. Some 96.0% of infertile couples in population C were ready to undergo PGD for aneuploidies. PMID- 9740455 TI - Endothelium-derived prostanoids reduce 5-hydroxytryptamine-induced contraction in the human uterine artery. AB - The contribution of endothelium-linked mechanisms to the contraction induced by 5 hydroxytryptamine (5-HT) was investigated in the isolated human uterine artery. 5 HT contracted the uterine artery concentration-dependently. Removal of the endothelium or treatment with the cyclooxygenase inhibitor indomethacin potentiated the contractile response to 5-HT. The nitric oxide synthase inhibitor L-N(G)-monomethyl-arginine (L-NMMA) did not influence the contraction induced by 5-HT. Indomethacin did not affect the response to 5-HT in endothelium-denuded vessels. The 5-HT1 receptor agonist 5-carboxyamidotryptamine (5-CT) did not relax precontracted arteries. Removal of the endothelium did not change the response to 5-HT in the presence of the 5-HT(1B/D) receptor antagonist GR127935 and the 5 HT1A and 5-HT1B receptor antagonist -pindolol. The 5-HT1B receptor antagonist SB224289 did not affect the contraction induced by 5-HT. The results indicate that the 5-HT-induced contraction in the human uterine artery is accompanied by the release of an endothelium-derived relaxing factor (EDRF). This EDRF seems to be a prostanoid, probably prostacyclin (PGI2). The endothelium-linked mechanism seems to be mediated via a 5-HT1 receptor, but it is not possible to further classify the receptor subtype by the information obtained in this study. PMID- 9740456 TI - Novel vaginal danazol ring therapy for pelvic endometriosis, in particular deeply infiltrating endometriosis. AB - Danazol is routinely administered orally to inhibit ovulation and to treat pelvic endometriosis. However, recent evidence suggests that danazol can act directly on endometriotic tissue in vitro to inhibit DNA synthesis and induce apoptosis. Danazol was administered via the vagina in this study, using a vaginal ring drug delivery system containing 1500 mg of danazol. This therapy was effective for treatment of pelvic endometriosis, especially for deeply infiltrating endometriosis, resulting in a cure of dysmenorrhoea and tenderness in the cul-de sac within 3 months, and of induration or nodularity in the cul-de-sac within 7 months. Moreover, conception was possible during insertion of the vaginal ring in 17 out of 31 infertile women with deeply infiltrating endometriosis, and in two out of eight infertile women with ovarian endometriotic cysts not adhering to the cul-de-sac and without deeply infiltrating endometriosis. Serum danazol concentrations, high during oral daily 400 mg danazol therapy, but undetectable during vaginal danazol ring therapy, explain why ovulation and conception could occur during insertion of the vaginal danazol ring, and why general side-effects, which are often observed during oral danazol therapy, were not observed during vaginal danazol ring therapy. Danazol seems to be absorbed through the vaginal mucosa and reaches the deeply infiltrating endometriosis via diffusion. PMID- 9740457 TI - Peritoneal fluid concentrations of interleukin-8 in women with endometriosis: relationship to stage of disease. AB - There is increasing evidence that immunological mechanisms play a role in the pathogenesis and pathophysiology of endometriosis. It was therefore of interest to study interleukin-8 (IL-8), a chemokine, in the peritoneal fluid and peripheral blood of women undergoing laparoscopic procedures. The presence and concentrations of IL-8 in relation to endometriosis, infertility and abdominal pain were evaluated. Samples of peritoneal fluid (n = 49) and peripheral blood (n = 50) were obtained from 50 consecutive patients undergoing laparoscopic surgery for various gynaecological indications (abdominal pain, infertility, sterilization). IL-8 was present in the peritoneal fluid of most women (87%). The concentration of IL-8 in the peritoneal fluid was higher in women with endometriosis compared to women without (P = 0.02). This difference was more pronounced in early (stage 1) endometriosis (P = 0.001). IL-8 concentrations in the peritoneal fluid were also higher in women with early endometriosis compared to women with later stages of the disease (P = 0.003). Peripheral blood concentrations did not correlate with peritoneal fluid concentrations of IL-8 and/or the presence of endometriosis. We conclude that IL-8 is an important factor that may contribute to the pathogenesis of endometriosis possibly by promoting neovascularization. This information can be a guide in the development of new therapeutic approaches for the treatment of endometriosis. PMID- 9740458 TI - Serum pregnancy-specific beta1-glycoprotein before embryo transfer is related to endometrial thickness and to outcome prognosis in women undergoing in-vitro fertilization treatment. AB - We have previously observed the repeated presence of low but detectable amounts of the trophoblast marker pregnancy-specific beta1-glycoprotein (SP1) in the serum of some women undergoing in-vitro fertilization (IVF) treatment around the time of oocyte retrieval. The occurrence of these signals seemed to be restricted to a defined group of patients which also showed a lower pregnancy success rate in a preliminary study. To test our hypothesis we have analysed 173 consecutive cycles leading to an embryo transfer. Fifty-four cycles (31%) had a serum SP1 level of at least 0.1 ng/ml between days embryo transfer -5 and embryo transfer (group A). Five pregnancies were obtained in this group (pregnancy rate = 9.3%), while in group B, defined by the absence of detectable SP1 before embryo transfer (119 cycles), 36 ongoing pregnancies were achieved (30.3%). Ten of the 41 pregnancies were achieved in 33 first-time non-pregnant patients undergoing further attempts during the study period. Again the pregnancy rate was higher in the first-time group B women (9/23 versus 1/10 for group A). Patients tended to remain in their groups A or B, the latter being associated with a better immediate as well as subsequent chance for pregnancy. Group A cycles had a significantly lower endometrial thickness two days before oocyte retrieval than group B (P = 0.0011). We postulate that the presence of an unknown, maternal and progesterone- or follicle stimulating hormone-independent factor in some patients could stimulate tonic ectopic SP1 synthesis and at the same time negatively influence endometrial development. PMID- 9740459 TI - Uterine contractions at the time of embryo transfer alter pregnancy rates after in-vitro fertilization. AB - To investigate the possible consequences of uterine contractions (UC) as visualized by ultrasound (US) on in-vitro fertilization (IVF)-embryo transfer outcome, we studied prospectively 209 infertile women undergoing 220 cycles of controlled ovarian stimulation. Inclusion criteria were age < or = 38 years, a morphologically normal uterus, and at least three good quality embryos transferred. Just before embryo transfer, women underwent 5 min digital recordings of the uterus using US image analysis software for UC assessment. Plasma progesterone and oestradiol concentrations were measured. Four groups were defined according to UC frequency: < or = 3.0 (n = 53), 3.1-4.0 (n = 50), 4.1-5.0 (n = 43), and > 5.0 (n = 74) UC/min respectively. Patients, controlled ovarian hyperstimulation and embryology characteristics were comparable in all groups. A stepwise decrease in clinical and ongoing pregnancy rates as well as in implantation rates occurred from the lowest to the highest UC frequency groups (53, 36, 21; 46, 32, 20; 23, 19, 10; and 14, 11, 4%; P < 0.001). Plasma progesterone and UC frequency were negatively correlated (r = -0.34, P < 0.001). Direction of UC did not affect embryo transfer outcome. As this study was controlled strictly for confounding variables and UC were assessed objectively by a computerized system, its results indicate that high frequency UC on the day of embryo transfer hinder IVF-embryo transfer outcome, possibly by expelling embryos out of the uterine cavity. The negative correlation between UC frequency and progesterone concentrations supports the uterine relaxing properties of progesterone. PMID- 9740460 TI - Mid-trimester loss--appraisal of a screening protocol. AB - The main causes for mid-trimester loss are known. There is likely to be overlap with those of first trimester loss, but the proportions may be different. We wished to perform an aetiological survey in a large population of patients with a history of recurrent miscarriage, for possible explanations for their second trimester miscarriages. Database analysis of 636 patients attending a UK University Teaching Hospital dedicated miscarriage clinic between 1991 and 1996 revealed a 25% prevalence (n = 158) for second trimester miscarriage. Results from an investigative screening protocol were positive in 50% of cases: 33% (n = 52) tested positive for antiphospholipid syndrome (APS); 8% (n = 13) fulfilled strict criteria for cervical incompetence; there was a 4% prevalence of uterine anomaly; 3% for infection (n = 5) and 2% of patients (n = 3) proved to be hypothyroid. Importantly, dual pathology was found in 5% of patients with a history of second trimester miscarriage. As idiopathic mid-trimester loss is a diagnosis by exclusion, a high index of suspicion is required, as are modern diagnostic techniques. PMID- 9740461 TI - Medical management of interstitial ectopic pregnancy: a case report and literature review. AB - Recent reports describe successful treatment of interstitial ectopic pregnancies using methotrexate. While the number of reported cases is increasing, no consensus exists regarding the management of this complication of pregnancy. We present the successful use of combined systemic and direct intrasac injection of methotrexate for an interstitial pregnancy with the highest yet reported initial beta-human chorionic gonadotrophin concentration (102,000 mIU/ml). We also describe the use of Doppler ultrasound for monitoring treatment progression. Through a review of the current literature, we propose to facilitate management decisions and increase outcome success by summarizing previously reported treatment regimens and by describing enhanced parameters for patient selection and monitoring. PMID- 9740462 TI - Mifepristone in combination with methotrexate for the medical treatment of tubal pregnancy: a randomized, controlled trial. AB - In the search for a more potent alternative to a single i.m. injection of methotrexate for ectopic pregnancy, a randomized trial was organized. The efficacy of a combination of methotrexate and mifepristone was compared with methotrexate alone in the treatment of unruptured tubal pregnancies. The diagnosis of an unruptured tubal pregnancy was confirmed laparoscopically in 50 patients during a 2 year period. Women were randomized to receive a single i.m. injection of 50 mg/m2 methotrexate alone or a single dose of 600 mg oral mifepristone in combination with the same dose of methotrexate. Both treatment protocols were successful in achieving the resolution of unruptured ectopic pregnancy (18/25 in the methotrexate group and 22/25 in the combination group) following the initial intervention. A second injection was needed in four (16%) cases in the methotrexate group and in one (4%) case in the combination group. Overall, a complete resolution was achieved in 22/25 and 23/25 cases respectively. Unruptured ectopic pregnancy resolved faster in women given the combination of methotrexate and mifepristone compared to women given methotrexate only (P = 0.01). The effect of the methotrexate and mifepristone combination was more pronounced in women with higher human chorionic gonadotrophin concentrations. PMID- 9740463 TI - Unintended pregnancies in women delivering at 18 South American hospitals. NFP ECLAMC Group. Latin American Collaborative Study of Congenital Malformations. AB - Unintended pregnancies are accepted as associated with social, maternal and perinatal risks, but few data exist in South America. In a selected network of hospitals participating in the ECLAMC (Spanish acronym for Latin American Collaborative Study of Congenital Malformations), the frequency of unintended pregnancies was 49.8% in 5155 mothers of normal liveborns, as interviewed in the post-partum period (1992-1994). Compared with the intended pregnancy group, these mothers were more frequently multiparous, conceived easily, had a surprisingly higher mean maternal age, lower educational level, and Black ancestors. The frequency of mistimed pregnancies was the highest among primiparae. No adverse perinatal outcome could be found with regard to low birthweight (< 2500 g), prematurity (< 37 weeks), and early neonatal death. The rates of Caesarean delivery, twinning and sex ratio were similar in intended and unintended groups. Logistic regression analysis showed that maternal education could be a confounding factor associated with other maternal variables. The rate of unintended pregnancies in the present study is significantly higher than that described for other regions. Knowledge of the characteristics of women experiencing unintended pregnancies would allow proper public health strategies. PMID- 9740464 TI - A 3 year, prospectively-designed study of late selective multifetal pregnancy reduction. AB - The aim of our study was to evaluate the pregnancy outcomes of late selective multifetal reduction (MFPR). We performed a 3 year, prospectively-designed study in which 28 patients underwent MFPR at a mean gestational age of 20.2 +/- 3.9 weeks (range 14-29 weeks). The indications for MFPR included: multiple gestation (> or = 3) (57%), structural anomaly (29%), and chromosomal abnormality (14%). The procedure was performed using ultrasonographically-guided intracardiac injection of potassium chloride. The mean gestational age at delivery was 36.6 +/ 2.2 weeks (range 31-40 weeks). Nine patients (32%) delivered before 36 weeks of gestation. The mean birth weight was 2370 +/- 614 g (range 1510-3250 g). Discordancy was evident in four twins (14%), and intrauterine growth retardation in four pregnancies. One case (3.5%) presented with oligohydramnios, and one with pregnancy-induced hypertension. One case of late abortion due to passive cervical dilatation 4 weeks after the MFPR was observed. Procedure-related amnionitis followed by late abortion occurred in one case. A total of 57% of the patients delivered vaginally and 43% delivered by Caesarean section. We concluded that late selective MFPR is associated with favourable perinatal outcome. Late MFPR may facilitate the detection of structural and chromosomal anomalies prior to the procedure, and the accomplishment of selective reduction of the affected fetus. PMID- 9740466 TI - Conservative laparoscopic management of a large cornual ectopic pregnancy. AB - Cornual pregnancy traditionally has been treated with laparotomy and either cornual resection or hysterectomy. Recently, more conservative operations have been developed, and operative laparoscopy has provided yet another management option. This report describes the conservative management of a large cornual ectopic pregnancy and reviews the techniques and outcomes of conservative repair that have been described in the literature. PMID- 9740465 TI - An unexpected triplet heterotopic pregnancy after replacement of two embryos. AB - We report a case of a triplet heterotopic pregnancy consisting of an intrauterine monozygous twin pregnancy and a tubal pregnancy after replacement of only two embryos in an in-vitro fertilization cycle with donor spermatozoa. This case demonstrates that sonographic demonstration of two intrauterine pregnancies after transfer of two embryos does not exclude the presence of an ectopic pregnancy. As both heterotopic pregnancy and spontaneous monozygotic twinning are more frequent after the use of assisted reproductive techniques, this combination, although extremely rare, must be kept in mind, especially in older patients with pre existing tubal damage. PMID- 9740467 TI - Lower risks of adverse outcome in twins conceived by artificial reproductive techniques compared with spontaneously conceived twins. AB - The outcomes of twins conceived by 136 women after medical assistance (MA) such as ovulation induction with or without assisted reproductive techniques, and twins conceived spontaneously (SP) by 72 women were compared. All 208 women were monitored from < 20 weeks gestation; they all delivered at > or = 24 weeks gestation. The chorionicity of the placenta was diagnosed antenatally and confirmed after delivery. There were 10 perinatal deaths; the physical and neurological status of the remaining 406 infants was assessed at 1 year of corrected age. There were no differences in gestational age at birth, the birth weights of the larger and smaller twins, the birth weight discordance, or the incidence of life-threatening major malformations between groups. Adverse infant outcomes, such as death, cerebral palsy and mental retardation occurred in nine (3.3%) of 272 MA twins compared with 12 (8.3%) of 144 SP twins (P < 0.05). The placenta was monochorionic in only three (2.2%) of 136 MA twin pregnancies compared with 41 (57%) of 72 SP twin pregnancies (P < 0.001). Of the 21 infants with adverse outcomes, nine had monochorionic placentas. Thus, the risk of an adverse outcome was 2.8-fold higher (95% confidence interval (CI) 1.2-6.4) in monochorionic twins than in dichorionic twins (10 versus 3.7%; P < 0.05). There was no difference in the incidence of adverse infant outcomes between SP (4.8%) and MA (3.4%) twins with dichorionic placentas. These findings suggest that ovulation induction in itself was not associated with an adverse outcome of twin pregnancies. The lower frequency of monochorionic placentas in MA twins may have been responsible for the lower risk of an adverse outcome in MA twins. PMID- 9740468 TI - Health and development of children born after oocyte donation compared with that of those born after in-vitro fertilization, and parents' attitudes regarding secrecy. AB - The health, growth and development of a cohort of children (n = 59) aged 6 months to 4 years and born after oocyte donation (OD) was compared with that from a group of children born after in-vitro fertilization (IVF) (n = 126). The study was performed by questionnaire, and the response rate was 100% in the OD group and 95% in the IVF group. All OD children were healthy. Three IVF children had a neurological disorder. Surgical intervention had been carried out in 8% of the OD and 13% of the IVF children. Height and weight development were normal, and eating and sleeping disorders were uncommon in both groups of children. The IVF mothers more often expressed concern about the child's behaviour than did the OD mothers. Thirty-eight percent of the OD parents and 60% of the IVF parents intended to tell the child about the nature of its conception (P < 0.01). Although oocyte recipients appear to have more complications during their pregnancies than conventional IVF patients, the general health status of OD children aged <5 years is at least as good as that of IVF children. Growth and development in both groups of children is similar to that of the general population. PMID- 9740469 TI - Demographic evaluation of the fertility of aluminium industry workers: influence of exposure to heat and static magnetic fields. AB - A demographic analysis of the fertility of French aluminium industry workers was performed in order to evaluate the potential effects on male fertility of occupational exposure to heat and static magnetic fields occurring in certain workshops. Two groups of aluminium workers were studied: one group of 692 potroom workers exposed to heat and to static magnetic fields, and a control group of 588 workers from the same plants, who had not been exposed to these factors. The birthrate was significantly higher in the 'exposed' group than in the 'control' group. The relative birthrate ratio ('exposed' versus 'control') was 1.1 (P < 0.001). These results do not show any decrease in the fertility of potroom workers exposed to heat and static magnetic fields, when compared to other workers in the aluminium producing industry. PMID- 9740470 TI - Recruitment and screening policies and procedures used to establish a paid donor oocyte registry. AB - We have reviewed the demographic characteristics of, and report abnormalities noted in, the de-novo growth and development of a paid oocyte donation programme. The personal profiles of all prospective oocyte donors were reviewed. Acceptance or rejection of candidates was based upon screening the results of medical, genetic and psychological testing. A total of 603 candidates initially responded to our advertisement. From this pool, 313 individuals were considered suitable and contacted by telephone. Following further conversation, 176 women were scheduled an entry interview. On completion of the formal screening process, 17.6% (n = 31) of those actually interviewed were denied entry. Thus, from the initial interested parties, only 23% of women wishing to participate in oocyte donation were considered suitable candidates. Given the high attrition rate, we concluded that the need for rigorous and thorough medical, psychological and genetic testing is mandatory for the establishment of a donor registry. Furthermore, professional counselling of prospective donors with respect to the results of tests and the implications of test results with respect to their future medical and reproductive health, are important parts of providing comprehensive care. PMID- 9740471 TI - Male infertility update. The ESHRE Capri Workshop Group. PMID- 9740472 TI - The value of maternal serum creatine kinase in the diagnosis of ectopic pregnancy. PMID- 9740473 TI - Beta2 glycoprotein I-dependent anticardiolipin antibodies in recurrent miscarriage. PMID- 9740474 TI - TESE and the distribution of spermatogenesis in the testicles of azoospermic men. PMID- 9740475 TI - Cooling rates in embryo cryopreservation. PMID- 9740476 TI - Clinical andrology today. PMID- 9740477 TI - Effects of exercise on natriuretic peptides and cardiac function in man. AB - We evaluated cardiac function and the plasma levels of atrial (ANP) and brain (BNP) natriuretic peptides during bicycle (B) and hand-grip (HG) exercises in eight healthy males. Each test was preceded by a control protocol in resting conditions. Left ventricular (LV) function (echocardiography) was evaluated during both exercises. Atrial function was assessed only during HG. Plasma ANP significantly increased during B (+236%) and HG (+77%), while there was a significant trend towards higher plasma BNP levels during B (+41%) and HG (+30%) than during the corresponding control tests. Plasma ANP correlated with heart rate in both tests, with left atrial volume, pulmonary vein flow systolic fraction and mitral flow E/A ratio in HG; BNP in both test correlated with LV dimensions and function. These data suggest that during exercise the cardiac release of ANP and BNP is differently regulated and related to changes in left atrial and LV function, respectively. PMID- 9740478 TI - Humoral response in patients with chronic heart failure. AB - AIM: Correlation of five humoral markers with laboratory, echocardiographic and right heart catheterization parameters in patients with chronic heart failure. STUDY POPULATION: 29 patients, heart failure NYHA II and III, ejection fraction below 40% with coronary artery disease or dilated cardiomyopathy. METHODS: evaluation of thromboxane, prostaglandin F (PGF), tumor necrosis factor (TNF) alpha, endothelin-1 and big endothelin rest levels and their correlation with: (1) laboratory parameters: Sodium, urea, creatinine, fibrinogen, (2) chest X-ray: cardiothoracic index (CTI), pulmonary congestion, (3) right heart catheterization parameters at rest, hand-grip and bicycle ergometry: mean pulmonary artery pressure (AP), wedge pressure (WP), systemic and pulmonary vascular resistance (SVR, PVR) and cardiac index (CI), (4) echocardiographic parameters at rest, hand grip and bicycle ergometry: end-diastolic volume (EDV), end-systolic volume (ESV), ejection fraction (EF), mitral flow E/A, filling period of left ventricle and time of duration of mitral regurgitation. RESULTS: No correlation was found between thromboxane, prostaglandin F and tumor necrosis factor alpha with the above mentioned parameters. Endothelin-1 level correlated with E/A, PVR and MPA at rest and at hand-grip. Big endothelin level correlated with EDV and ESV, AP, WP and SVR at rest and at both types of exercise. The highest correlation was between big endothelin and rest AP (r=0.79), rest WP (r=0.78) and CTI (r=0.58), all P<0.01. CONCLUSIONS: Big endothelin and partly endothelin-1 levels showed a close correlation with some parameters used for the evaluation of chronic heart failure severity. PMID- 9740479 TI - Suppressing sympathetic activation with clonidine on ventricular arrhythmias in congestive heart failure. AB - This randomized, double-blind, placebo-controlled study examines the effects of clonidine (a centrally acting sympathoinhibitor) on ventricular arrhythmias in 35 patients with congestive heart failure (CHF) by using the 24-h ambulatory electrocardiographic recording. After baseline examination and Holter recording, patients were balanced and 18 patients were randomized to clonidine group and 17 patients to placebo group. After four weeks of clonidine (given as a transdermal patch) or matching placebo therapy, a second Holter recording was obtained. The placebo group showed no change in the frequency of ventricular arrhythmias whereas the clonidine-treated group showed a significant decrease in the frequency of ventricular premature beats by 68% (P<0.01), couplets by 63% (P<0.01) and episodes of non-sustained ventricular tachycardia by 60% (P<0.05). Clonidine also decreased heart rate and arterial blood pressure, but left ventricular ejection fraction was slightly improved. It is concluded that sympathetic suppression with clonidine reduces the frequency of ventricular arrhythmias in patients with CHF, which suggests that sympathetic activation plays a role in arrhythmogenesis in these patients. PMID- 9740480 TI - Valsartan in heart failure patients previously untreated with an ACE inhibitor. AB - OBJECTIVE: To evaluate the effect on cardiac hemodynamic parameters of valsartan in patients with chronic stable congestive heart failure previously untreated with ACE inhibitors. METHODS: After a 2 to 4 week run-in period, 116 adult outpatients were randomized to receive valsartan 40, 80 or 160 mg twice daily, the ACE inhibitor lisinopril 5/10 mg once daily, or placebo. At baseline and after 28 days of treatment, cardiac hemodynamic parameters were measured. Tolerability was assessed by adverse events and by any changes in systolic or diastolic blood pressure, body weight, heart rate, and routine laboratory parameters. RESULTS: For the 12 hour time point (trough), all doses of valsartan reduced mean pulmonary capillary wedge pressure (statistically significant for valsartan 40 mg and 160 mg), decreased systemic vascular resistance (statistically significant for all three valsartan doses and for lisinopril at peak and trough), and increased cardiac output (statistically significant for all three valsartan doses at peak, and for 80 and 160 mg at trough). There were no clinically relevant effects on any safety parameters. CONCLUSIONS: Valsartan has beneficial effects on cardiac hemodynamics, and is generally well tolerated in patients with congestive heart failure not taking ACE inhibitors. PMID- 9740482 TI - Clinical significance of QT dispersion after exercise in patients with previous myocardial infarction. AB - To evaluate the clinical significance of QT dispersion after exercise in patients with previous myocardial infarction, QT dispersion (QTd) and corrected QTd (QTcd) were assessed with standard 12 leads electrocardiogram in 90 patients with previous myocardial infarction and 30 healthy persons before and 3 min after a treadmill exercise test. In addition, 24 h ambulatory electrocardiogram and echo cardiography were examined in all the subjects studied. Patients were followed up for 37.25 +/- 10.71 months. The results showed that there were no significant differences in the QTd and QTcd between the patients and the controls before exercise (36.11 +/- 13.42 ms versus 34.81 +/- 12.32 ms, P>0.05, 41.22 +/- 13.49 as versus 39.91 +/- 13.56 ms, P>0.05). Compared with those before exercise, QTd and QTcd were significantly increased in the patients 3 min after the exercise test (36.11 +/- 13.42 ms versus 47.20 +/- 14.41 ms, P<0.01, 41.22 +/- 13.49 ms versus 59.57 +/- 18.90 ms, P<0.01), but not in the controls (34.81 +/- 12.32 ms versus 38.76 +/- 12.09 ms, P>0.05, 39.91 +/- 13.56 ms versus 43.27 +/- 17.77 ms, P>0.05). The incidences of abnormal contraction of the left ventricular wall, aneurysms, NYHA III class, >III class of Lown's ventricular arrhythmia classification and cardiac sudden death were significantly higher in group A with QTcd >50 ms than that of group B with QTcd <50 ms (P<0.01). These findings indicate that the increased QT dispersion after exercise in 12 standard leads electrocardiogram might be associated with high incidences of sudden cardiac death and ventricular arrhythmia in the patients with previous myocardial infarction. PMID- 9740481 TI - Exercise performance in patients with dilated cardiomyopathy: relationship to resting left ventricular function. AB - Relationship between maximal exercise tolerance and resting indexes of left ventricular systolic and diastolic function were evaluated in 35 men, aged 55.1 +/- 10.4 years, with dilated cardiomyopathy. Clinical diagnosis of dilated cardiomyopathy was confirmed with M-mode echocardiography (M-mode echocardiographic end-diastolic dimension >55 mm, fractional shortening <25%, increased E point septal separation). Coronary angiography was considered mandatory for exclusion of patients with coronary artery disease. Patients with mitral regurgitation (> or =grade 2) and rhythm other than sinus were excluded. According to the functional classification of New York Heart Association 6 patients were in class I, 11 in class II, 12 in class III and 6 in class IV. Left ventricular ejection fraction (LVEF), stroke volume (SV) and left ventricular end diastolic pressure (LVEDP) were measured with contrast angiography. Peak early (VE) and late (VA) transmitral filling velocities and their ratio (E/A), isovolumetric relaxation time (IRT) and deceleration time (DT) were computed from pulsed wave Doppler echocardiograms. On completion of all resting measurements, patients underwent symptom limited upright treadmill exercise testing using a modified Naughton protocol and maximal exercise performance metabolic equivalent work load (NETS) was calculated from the speed, incline and length of time at the stage using standard tables to make interpatient comparisons. Significant correlation has been found between NYHA class and METS (r= -0.77, P<0.001). However NYHA class II and NYHA class III patients were found to have similar METS (P=0.317). Patients were further divided into two groups on the basis of exercise data. Group I consisted of 22 patients with relatively preserved exercise tolerance (> or =4 METS) and Group II included 13 patients with impaired exercise tolerance (> or =4 METS). This arbitrary classification was based upon previously described survival differences in these two groups. There were no differences between two groups in terms of age, gender distribution (all were male), heart rate and arterial blood pressure. LVEF, LVEDP, stroke volume, VE, VA, E/A, IRT and DT were also similar between two groups. Strong positive correlation was observed between LVEDP and VE (r=0.74) while IRT and VA negatively correlated with LVEDP (r= -0.77 and r= -0.81 respectively) but neither of resting indexes of left ventricular systolic and diastolic function showed significant correlation with METS and exercise duration. PMID- 9740483 TI - Cardiac autonomic impairment and early myocardial damage involving the right ventricle are independent phenomena in Chagas' disease. AB - Cardiac autonomic impairment and right side heart failure are prominent features in patients with Chagas' disease, but no causal relationship between these phenomena has been disclosed and the pathophysiology of such manifestations is unclear. Aim of study was to assess the cardiac autonomic control and biventricular function in chagasic patients in early stages of the disease, using radionuclide angiography, Valsalva manoeuvre, head-up tilt and baroreflex sensitivity evaluation. Thirty-one chagasic patients with no clinical signs of Chagas' heart disease-16 in the indeterminate phase and 15 with sole organic digestive involvement-were studied, and results compared with those obtained in 14 normal volunteers. No significant differences were observed among the three groups, in regard to any systolic or diastolic parameter of LV function, including ejection fraction, peak ejection and filling rates and correspondent times, time to end-systole, and the standard deviation of phase values. The indeterminate and digestive groups of chagasics had significantly lower right ventricular ejection fraction (45.7 +/- 6.3 and 46.2 +/- 10.1 respectively) and peak ejection rate (respectively 2.8 +/- 0.6 and 2.9 +/- 0.6) and higher right ventricular phase standard deviation (22.4 +/- 5.9 and 20.1 +/- 5.6 degrees, respectively), as compared with the control group (53.6 +/- 4.3, 3.5 +/- 0.5, and 15.8 +/- 3.8 respectively for right ventricular ejection fraction, peak ejection rate and phase standard deviation). No significant differences were found between the results of autonomic evaluation in the control and indeterminate groups of chagasic patients. The group of digestive disease patients showed abnormally lower Valsalva ratio (1.5 +/- 0.15), baroreflex sensitivity (8.85 +/- 2.05 ms/mmHg) and parasympathetically-dependent heart rate response to tilt (8.85 +/- 8.42 beats/mm) and higher Valsalva delay (15.67 +/- 1.35 s) values, compared with the control group (respectively 1.85 +/- 0.49, 20.23 +/- 12.66 ms/mmHg, 21.61 +/- 5.77 beats/mm and 10.1 +/- 2.5 s). Thus, cardiac autonomic impairment is a prominent feature in chagasic patients with the digestive but not the indeterminate form of Chagas' disease. It bears no causative relationship to the early myocardial damage that is apparent only regarding right ventricular function, in both groups of patients. Early right ventricular dysfunction is a likely mechanism for the marked predominance of systemic over pulmonary congestion when heart failure supervenes in patients with Chagas' disease. PMID- 9740484 TI - Association between coronary angiograms and cardiac events--a prospective 3-year follow-up. INTACT-Investigators. International Nifedipine Trial on Antiatherosclerotic Therapy. AB - The correlation between extent and severity of coronary artery disease as documented by quantitative coronary angiography and the incidence of cardiac events within 3 years was analyzed from a prospective study. In 73 out of 419 patients, 89 events occurred comprising 10 cardiac deaths, 15 non-fatal myocardial infarcts, 26 cases of unstable angina, and 38 coronary revascularization procedures (bypass graft operation or angioplasty). The incidence of any event correlated with the baseline number of all stenoses and high-grade stenoses (> or =20% and > or =50% diameter stenosis, respectively) (P<0.05). With respect to specific events, non-fatal myocardial infarcts and revascularization procedures were correlated with the number of all stenoses (P<0.05), but not with high-grade stenoses. Specification of coronary arteries revealed correlation of non-fatal myocardial infarcts and revascularization procedures with the number of high-grade stenoses in the left anterior descending artery. Finally, baseline left ventricular ejection fraction was found to be lower in patients who died of cardiac causes than in the remaining patients (49 +/- 10% vs. 67 +/- 13%; P<0.001). In conclusion, the total coronary stenosis burden seems to predict the incidence of subsequent cardiac events even better than the number of high-grade stenoses. Only in the left anterior descending artery high-grade stenoses seem to cause myocardial infarcts within a relatively short period of time justifying short-term revascularization in these patients. PMID- 9740485 TI - The Brugada syndrome in a Chinese population. AB - Sudden cardiac death has been reported in patients with a unique electrocardiographic (ECG) abnormality showing right bundle branch block and ST segment elevation in the precordial leads. This syndrome was first described by Brugada and Brugada and has not been previously described in a Chinese population. We report here the first three cases in Singapore. The first patient was a 49-year-old man who presented with syncope, associated with generalized convulsions. The second patient was a 25-year-old man who complained of palpitations but no syncope. The third patient was a 77-year-old man who presented with recurrent episodes of syncope and collapsed with ventricular fibrillation. All patients had no past cardiac or drug history of note. The neurological examination and investigations were normal. All three patients showed a unique right bundle branch block pattern with ST segment elevation in leads V1-3. The echocardiogram and 24-h ambulatory ECG monitoring, were normal. Single vessel disease was present in the third patient. Electrophysiological studies performed in all three patients were able to induce ventricular fibrillation. The patient with resuscitated cardiac death underwent an implantable cardioverter defibrillator implantation. The importance of this syndrome is that the recognition of the unique ECG pattern enables early identification and treatment of these patients. PMID- 9740487 TI - A case of primary cardiac lymphoma: utility of serum soluble interleukin-2 receptor for noninvasive diagnosis. AB - A 70-year-old woman was referred to us for the investigation of a tumor mass in the right sided of the heart. LDH and soluble interleukin-2 receptor (sIL2r) were elevated. She died one day after the initiation of radiation therapy because of intractable cardiogenic shock. Primary cardiac lymphoma was identified at autopsy. Measurement of serum sIL2r lead us to the antemortem diagnosis of the disease. PMID- 9740486 TI - Fatty acid beta-oxidation deficiency masquerading as fulminant myocarditis. AB - We present a 9-month-old child presenting with acute cardiomyopathy and fever following a viral illness. He was diagnosed to have acute myocarditis, was proposed for an external hemodynamic assistance but died of ventricular tachycardia. Post-mortem data revealed a very-long-chain acylcoenzyme A dehydrogenase deficiency. Our report raises awareness on the interest for preserving tissue samples and for performing a metabolic screening in acute childhood cardiomyopathy. PMID- 9740488 TI - Recurrent restenosis following stent and rotational atherectomy of coronary artery stenosis in Takayasu's arteritis. AB - We report a patient with Takayasu's arteritis who had recurrent restenosis following intracoronary bifurcation stenting of proximal left anterior descending and first diagonal arteries, and rotational atherectomy for in-stent restenosis. After all, the patient underwent coronary artery bypass grafts (CABG) and has remained asymptomatic during 3 months without damaging myocardium. We suggest that endoluminal stenting or rotational atherectomy may be an alternative treatment for the patients with coronary artery stenosis due to active Takayasu's arteritis as a therapy to postpone CABG. PMID- 9740489 TI - Long term follow-up in a case of anomalous origin of the left coronary artery from the pulmonary artery. AB - An adult patient with anomalous origin of the left coronary artery from the pulmonary artery had no serious cardiac event for 19 years after closing the shunt from the aorta to the pulmonary trunk by ligation of the left main coronary artery. Although one-coronary arterial perfusion is considered to have worse prognosis than two-coronary arterial system, this patient proved to have an unexpectedly good long-term prognosis. PMID- 9740491 TI - Fluoxetine-induced QTU interval prolongation, T wave alternans and syncope. PMID- 9740490 TI - An unusual case of myocarditis. AB - A 25-year-old woman was admitted to our center for left heart failure. The clinical, electrocardiographic and bioptic findings suggested an acute lymphocytic myocarditis. One month later, paroxysmal hypertension and high urinary excretion of noradrenaline revealed the presence of a pheochromocytoma. PMID- 9740492 TI - Cisapride can make Prinzmetal angina worse. PMID- 9740494 TI - Impaired glucose tolerance: a risk factor for type 2 diabetes and cardiovascular disease. PMID- 9740493 TI - Unexpected recurrence of sinus rhythm in chronic atrial fibrillation due to sick sinus disease. AB - A 60-years old patient with symptomatic sick sinus disease was implanted a dual chamber pacemaker that did well during the following 13 years. Thereafter, the pacemaker had to be explanted because of a systemic infection, with the patient in constant chronic atrial fibrillation in the meantime. After another asymptomatic year, 6 arrhythmogenic syncopes occurred, apparently due to pre automatic pauses prior to spontaneous conversions to sinus rhythm. Subsequently, the patient was implanted a VVI pacemaker, whereupon he did well henceforth. This case demonstrates the possibility of recurrence of sinus rhythm even after long standing chronic atrial fibrillation. PMID- 9740495 TI - Impaired glucose tolerance: what are the clinical implications? AB - Impaired glucose tolerance (IGT) was standardized in 1979 by the National Diabetes Data Group and the World Health Organization as a risk factor for type 2 diabetes, replacing groups such as 'borderline' and 'chemical' diabetes. IGT was defined by a blood/plasma glucose value 2 h after a 75 g glucose load that was clearly abnormal but did not convey a risk of microangiopathy in those with non diabetic fasting blood/plasma glucose levels. IGT is not uncommon, having a prevalence of 2-25% in adults. Determinants include age, obesity (total and central), family history of type 2 diabetes, physical inactivity and triglyceride levels. The main clinical significance of IGT is: (1) as a risk factor for type 2 diabetes, with 20-50% of individuals developing type 2 diabetes over 10 years; (2) as a risk factor for cardiovascular disease (CVD); and (3) as a component of the metabolic syndrome. IGT can be treated and this may prevent or delay progression to type 2 diabetes, though the effect of treatment on the risk of CVD is unknown. PMID- 9740496 TI - Dysglycaemia: a cardiovascular risk factor. AB - Patients with diabetes have a 2-fold higher risk of developing cardiovascular disease than non-diabetic individuals. Moreover, recent epidemiologic studies have shown that this risk rises with the degree of hyperglycaemia, so that diabetic patients with poorly controlled glucose levels have a higher risk of cardiovascular disease than those with well-controlled glucose levels. Thus, in patients with diabetes, glucose level appears to be a continuous risk factor for cardiovascular disease. Several epidemiologic studies also suggest that this relationship is not confined to the diabetic range; non-diabetic levels of fasting and postprandial hyperglycaemia, that may even be lower than those associated with impaired glucose tolerance, are also associated with an increased risk of cardiovascular disease. Evidence is therefore accumulating that dysglycaemia (i.e. raised glucose levels above some low, as yet undefined, threshold) is a continuous risk factor for cardiovascular disease. This relationship is similar to that of smoking, blood pressure and dyslipidaemia to cardiovascular risk. Whether glucose lowering in diabetic or non-diabetic individuals will prevent cardiovascular disease remains to be determined. PMID- 9740497 TI - Implications of blood glucose, insulin resistance and beta-cell function in impaired glucose tolerance. AB - Insulin secretion is stimulated by ingestion of food. The combination of hyperinsulinaemia plus hyperglycaemia effectively promotes glucose uptake by the liver and by peripheral tissues, such as muscle and fat cells, and suppresses hepatic glucose output. These simultaneous processes maintain normal glucose homeostasis in a co-ordinated fashion. Type 2 diabetes mellitus is associated with impaired insulin in target tissues due to insulin resistance and/or insulin deficiency. At first, increased insulin secretion overcomes insulin resistance, but ultimately this fails, leading progressively to increased blood glucose levels. Individuals pass through a phase of impaired glucose tolerance (IGT) and increased fasting plasma glucose levels (IFG) before developing overt type 2 diabetes. Therefore, IGT/IFG is a dysglycaemic state that is intermediate between normal glucose tolerance and diabetes. In this article, we discuss the relative importance of hyperglycaemia, insulin resistance and beta-cell function in the development of glucose intolerance, taking the new diagnostic criteria into consideration. New recommendations from the Expert Committee on the Diagnosis and Classification of Diabetes Mellitus are discussed where appropriate. PMID- 9740498 TI - Assessing the potential for alpha-glucosidase inhibitors in prediabetic states. AB - Type 2 diabetes often has an insidious onset with hyperglycaemia being present for many years before diagnosis is made. It is a progressive disease, due in part to loss of beta-cell function, with the reduction in function probably commencing 10-12 years prior to diagnosis and being aggravated by increasing fasting plasma glucose levels. Earlier intervention in those at risk from type 2 diabetes, aimed at minimizing hyperglycaemia, may prevent or delay overt diabetes and the associated development of micro- and macrovascular disease. Six-year follow-up data from the UK Prospective Diabetes Study, confirm that sulphonylurea, metformin and insulin therapy can reduce hyperglycaemia in individuals with type 2 diabetes. Although none of these agents prevent the subsequent progressive increase in fasting glucose levels, preliminary results with acarbose show that fasting plasma glucose levels can be maintained over 1 year of therapy. Three large-scale studies are currently investigating whether treatment with acarbose at an earlier stage of the disease process, in subjects with varying degrees of glucose intolerance, may be beneficial in helping to prevent or delay the onset of diabetes. PMID- 9740499 TI - Postprandial hyperglycaemic state: importance and consequences. PMID- 9740500 TI - Intermediate-density lipoproteins, diabetes and coronary artery disease. AB - The results of various studies suggest that hypertriglyceridaemia is associated with an increased risk of coronary artery disease. It is unclear, however, which particular triglyceride (TG)-rich lipoproteins contribute to the risk. Different types of TG-rich lipoprotein differ in function, composition, size and density. TG-rich lipoproteins in the range Svedberg flotation (Sf) 12-60 have been shown to be associated with angiographic severity in both diabetic and non-diabetic individuals. A study in people with type 2 diabetes found that those with moderate coronary artery disease had higher levels of both Sf 12 60 and Sf 60 400. Multivariate analysis showed that this association was independent of both low (LDL)- and high-density lipoprotein (HDL). The association was not seen in patients with severe coronary artery disease, suggesting that these lipoproteins may only be involved in the early stages of atherogenesis. Further research has indicated that the risk correlates positively to the postprandial levels of apolipoprotein B48 in the Sf 20-60 fraction. This suggests that elevated levels of chylomicron remnants are involved in progression of coronary artery disease. PMID- 9740501 TI - Asymptomatic hyperglycaemia and early atherosclerotic changes. AB - Ultrasonographic scanning of carotid arteries allows non-invasive detection of atherosclerotic changes. This technique has been used to investigate changes in the thickness of the intimal plus medial (IM) complex, in patients with type 2 diabetes, type 1 diabetes and those in the pre-diabetic state of impaired glucose tolerance (IGT). IM thickness (IMT) increases with age, but this process was found to be considerably accelerated in patients with type 2 diabetes. In addition, IMT was significantly greater in patients with cerebral lacunar infarctions, and in those with detectable coronary artery stenosis. A study in patients with type 1 diabetes found that IMT correlates with duration of diabetes as well as age. The correlation with duration of diabetes suggests that hyperglycaemia contributes to the progression of atherosclerosis. IMT was also found to be increased in individuals with hyperinsulinaemic IGT, compared with control individuals with normal glucose tolerance. These results suggest that even relatively small increases in postprandial blood glucose levels can lead to increases in IMT and, hence, increased risk of cardiovascular disease. Further analysis revealed a correlation between hyperinsulinaemia (i.e. insulin resistance) and increased IMT. These results provide a clear rationale for the therapeutic use of alpha-glucosidase inhibitors, such as acarbose, which attenuate postprandial hyperglycaemia-induced hyperinsulinaemia. PMID- 9740502 TI - The clinical importance of postprandial glucose. AB - Atherosclerotic lesions develop over a long period of time and result from complex changes in the arterial wall. Although these changes are not fully understood, there is much evidence to suggest that elevated plasma glucose levels contribute to the development of atherosclerotic lesions. Many studies have shown that there is a strong correlation between elevated plasma glucose levels and the risk of developing cardiovascular disease. Effects of glucose on the arterial wall include immediate effects, which occur rapidly in response to elevated plasma glucose levels, and long-term effects, which result from non-enzymatic glycosylation of various proteins. These adverse effects of elevated plasma glucose levels suggest that tight control of blood glucose levels in patients with diabetes could possibly reduce the risk of cardiovascular complications. This is borne out by the results of clinical studies in patients with type 1 diabetes. Therapy to reduce blood glucose levels may also be appropriate in individuals with impaired glucose tolerance, as this condition is associated with postprandial hyperglycaemia and a significant risk of developing cardiovascular disease. PMID- 9740503 TI - Postprandial hyperglycaemia and alpha-glucosidase inhibitors. AB - Fasting blood glucose level is usually used to diagnose diabetes, but is not a good predictor of postprandial hyperglycaemia, which is a more accurate measure of the metabolic defect underlying type 2 diabetes. Postprandial blood glucose levels may be elevated while fasting levels are normal, constituting an early stage in type 2 diabetes that can be termed 'postprandial diabetes'. Prevention of postprandial hyperglycaemia is important, as it is implicated in the development of macro- and microvascular complications associated with diabetes. The risk of cardiovascular disease is higher in individuals with postprandial hyperglycaemia, even without diabetes, than in individuals with normal postprandial blood glucose levels. Furthermore, postprandial hyperglycaemia is implicated in the development of type 2 diabetes. Even modest postprandial hyperglycaemia may lead to beta-cell dysfunction. Agents that reduce postprandial hyperglycaemia have a key role in the treatment of type 2 diabetes and pre diabetic states. Most anti-diabetic agents that are currently available reduce fasting blood glucose levels, but have little impact on postprandial glycaemic excursions and thus do not normalize postprandial hyperglycaemia. However, new agents that control postprandial hyperglycaemia have been developed, for example, the alpha-glucosidase inhibitor acarbose. Such agents have a potential to reduce the progression of diabetes as well as macro- and microvascular complications. PMID- 9740504 TI - Melanoma-associated antigens recognized by cytotoxic T lymphocytes. AB - During the last 7 years significant progress has been made in the identification of melanoma-associated antigens recognized by cytotoxic T lymphocytes (CTL). These antigens belong to three main groups: cancer/testis-specific antigens (MAGE, BAGE, GAGE, PRAME and NY-ESO-1), melanocyte differentiation antigens (tyrosinase, Melan-A/MART-1, gp100, TRP-1 and TRP-2), and mutated or aberrantly expressed antigens (MUM-1, CDK4, beta-catenin, gp100-in4, p15 and N acetylglucosaminyltransferase V). In this review we have summarized the available data concerning the characterization of melanoma-associated antigens, focusing on their immunogenic and protective properties. The development of a strong immune response to differentiation antigens is limited by the existence of tolerance to these "self"-antigens, permitting the involvement of only T cells with low affinity T-cell receptors. Among the melanoma differentiation antigens, only gp100 has been shown to be a tumor regression antigen. The cancer/testis-specific antigens such as MAGE and PRAME should potentially be highly immunogenic antigens. They contain several potential HLA class I binding epitopes and are present only in the testes, which are not accessible to the cells of the immune system owing to the lack of direct contact with the immune cells and the lack of HLA class I expression on the surface of germ cells. But only two patients have been found who responded to these antigens in vivo, indicating their genuinely low immunogenicity. A comparison of the predicted secondary structures of these two groups of antigens (cancer/testis-specific and differentiation antigens) revealed enrichment of long alpha-helical stretches in the cancer/testis-specific antigens. We hypothesize that such highly organized stable structures could, first, reduce denaturation of the protein and, thus, ubiquitinylation as a degradation signal, and, second, diminish the efficiency of the protein unfolding - a necessary step in the proteolytic cleavage by proteasomes. High structural stability could therefore be responsible for the low immunogenicity of these proteins. In this case, modifications decreasing the stability of these proteins might be a means of improving the immune response to these potentially therapeutically useful antigens. PMID- 9740505 TI - Diagnosis of congenital Toxoplasma gondii infection by polymerase chain reaction (PCR) on amniotic fluid samples. The Norwegian experience. AB - As part of a screening project for detection of Toxoplasma gondii infection among pregnant women in Norway, nested polymerase chain reaction (PCR) aimed at the detection of T. gondii in amniotic fluid samples was included in the diagnostic routine. The results were compared with the routine criteria for congenital infection: i) T. gondii detected in amniotic fluid or cord blood by mouse inoculation, ii) specific IgM or IgA in serum collected after birth, and/or iii) specific IgG persisting beyond one year of age. The PCR was based on the B1 gene with an internal control gene amplified together with the B1 gene. One hundred and two amniotic fluid samples collected during pregnancy and/or at delivery from 67 pregnant women with serological evidence of primary T. gondii infection were available for examination by both B1-PCR and mouse inoculation. Six samples were positive and 86 samples were negative by both methods (90% concordance). One sample was mouse inoculation positive and B1-PCR negative while nine samples were B1-PCR positive and mouse inoculation negative, of which five were associated with four infants without proven infection. 59%, and 41% of samples associated with infected infants were positive by B1-PCR and mouse inoculation, respectively. The difference was mainly due to a lower detection rate by mouse inoculation after antiparasitic treatment. The specificity of B1-PCR was 94%. Even though B1-PCR performed on amniotic fluid samples did not detect all infected infants, it represented a valuable tool in addition to conventional methods in the diagnosis of congenital T. gondii infection. PMID- 9740506 TI - Pulmonary intravascular macrophages appear in rats after long-term administration of lipid emulsion and amino acid solution. An ultrastructural morphometric study. AB - Intravascular macrophages have rarely been seen in normal lungs of humans and rats, but in rats endotoxaemia has induced their presence. To study whether substrates used for parenteral nutrition could have a similar stimulatory effect on mononuclear phagocytes, rats were given lipid emulsion (n=5), amino acid solution (n=5), or isotonic saline (n=5) through central venous catheters for 3 weeks. Structural changes in the lung microvessels were evaluated using electron microscopy. The areal fraction of pulmonary intravascular mononuclear phagocytes was 19.6% (SD=8.2) in rats given lipid emulsion (p<0.05) and 8.2% (SD=8.2) in rats given amino acid solution n.s. compared to 2.4% (SD= 4.0) in rats given saline. The increase in areal fraction was mainly due to an increase in cell numbers. In rats given lipid emulsion the intravascular phagocytes were only slightly larger than in rats given saline, but had the morphological features of mature macrophages. The study demonstrates that lipid emulsion recruits pulmonary intravascular macrophages in rats, indicating a stimulatory effect on the mononuclear phagocyte system. The effect was less pronounced with amino acid solution. PMID- 9740507 TI - The value of autopsy from a clinical point of view. A survey of 250 general practitioners and hospital clinicians in the county of Sor-Trondelag, Norway. AB - An enquiry into clinicians' attitudes to autopsies was conducted by questionnaire sent to 166 general practitioners in the county of Sor-Trondelag in Norway and to 186 clinicians working at the University Hospital of Trondheim. It was considered especially important for us to include general practitioners. As the result of a government decision taken at the end of 1994, autopsies requested by general practitioners on patients dying outside hospitals have from January 1995 been covered by national health insurance. Answers were obtained from 250 doctors: 110 general practitioners and 140 hospital physicians. One hundred and seventy-nine (73.1%, n=245) felt that the possibility of having autopsies performed was of great importance in their daily work. Autopsy was considered to be a very important means of quality assurance in the health care system by 158 (66.4%, n=238). One hundred and two (41.2%, n=247) answered that improvements in medicine and technology during the last decades had not reduced the importance of autopsy. One hundred and twenty-two (81.3%, n=150) felt that especially computer tomography (CT) had reduced the value of autopsy. Among the general practitioners, 73 (68.9%, n=106) welcomed the opportunity to have non-forensic autopsies performed on patients who died outside hospitals. Our study showed differences in the attitudes of clinicians towards autopsies, but our results still indicate that the value of autopsy for furthering clinical knowledge is acknowledged by most clinicians. PMID- 9740508 TI - Diagnosis of pulmonary tuberculosis. Application of gen-probe amplified Mycobacterium tuberculosis direct test. AB - The purpose of this investigation was to evaluate the Amplified Mycobacterium tuberculosis Direct Test (AMTDT) for the diagnosis of pulmonary tuberculosis (TB). Six hundred and forty-six sputum samples were analysed by microscopy for acid-fast bacilli, by culture for mycobacterial growth, and by AMTDT for the presence of M. tuberculosis complex rRNA. If there were discrepant results, information as to whether the patient had a history of TB was obtained. The sensitivity was 85.7% (smear positive 100% and smear negative 71.4%, respectively) and the specificity was 96.8% compared with culture. By retesting, 12 of 13 false-positive samples could be divided into two major categories. The samples in one category were retest positive, had highly positive results, and the patients had previous culture-proven TB. The samples in the other category had less positive AMTDT results, were retest negative, and the patients were not notified as having TB. This investigation shows that AMTDT is a sensitive supplementary method for rapid detection of M. tuberculosis complex in respiratory specimens, that there is a risk of contamination which has to be monitored closely, and that positive samples should be retested. PMID- 9740509 TI - S-100 protein-positive dendritic cells in follicular lesions of the thyroid. AB - Little is known about the role of S-100 protein-positive dendritic cells in follicular lesions of the thyroid. In 36 cases of adenomatous goitre, 18 cases of follicular adenoma, and 7 cases of follicular carcinoma, we investigated the incidence and distribution of S-100 protein-positive dendritic cells using immunohistochemical staining. Four cases (10.5%) of adenomatous goitre, five cases (31.3%) of follicular adenoma, and 6 cases (85.7%) of follicular carcinoma contained S-100 protein-positive dendritic cells in the subcapsular area and/or the hyalinized stroma. Dendritic cells in the subcapsular area were observed only in lesions with thick capsules and not those with thin capsules. In most cases of follicular carcinoma, + + to + + + of the dendritic cells was seen together with a few lymphocytes and fibroblasts in the subcapsular areas. In contrast, cases of adenomatous goitre and follicular adenoma showed only a few dendritic cells without any inflammatory cells. Dendritic cells in follicular thyroid lesions may play some role in capsular formation or may be a secondary phenomenon due to capsular formation. PMID- 9740510 TI - Lack of active lung anaphylaxis in congenitally mast cell-deficient Ws/Ws rats sensitized with the nematode Nippostrongylus brasiliensis. AB - Ws/Ws rats are deficient in both mucosal- and connective tissue-type mast cells. To study the role of mast cells in active anaphylaxis, changes in vascular permeability in the trachea upon intravenous antigen challenge with Evans blue dye were examined in Ws/Ws, heterogenic Ws/+, and normal +/ + rats sensitized with the nematode Nippostrongylus brasiliensis. Antigen challenge resulted in fatal anaphylactic shock in some +/+ and Ws/+ rats, but not in Ws/Ws rats. Marked dye leakage developed within 30 min in the trachea of +/+ and Ws/+ rats, while Ws/Ws rats showed no substantial increases in the levels of vascular permeability. Ex vivo stimulation of sensitized lung fragments from +/+ animals with specific antigen induced significant releases of histamine and leukotriene (LT) C4, while sensitized Ws/Ws rat-lung fragments did not. In Ws/Ws rats, levels of nematode-specific IgE, IgG1 and IgG2a antibodies as well as levels of lung eosinophilia were not significantly different from those in +/+ rats. These results show that mast cell-deficient Ws/Ws rats fail to develop active anaphylaxis, and this is mediated probably by the lack of mast cell-derived mediators required for initiation of the reaction. PMID- 9740511 TI - Failure to detect Chlamydia pneumoniae in calcific and degenerative arteriosclerotic aortic valves excised during open heart surgery. AB - Chlamydia pneumoniae has been associated with atherosclerosis, although no causal association has been established. Employing culture and polymerase chain reaction in aortic valves with calcific and degenerative arteriosclerotic changes from 23 non-consecutive patients undergoing aortic valve replacement, C. pneumoniae was not detected in any of the valves. 19/22 patients had serological evidence of past infection with C. pneumoniae. Our findings do not provide supportive evidence for the hypothesis that C. pneumoniae is associated with calcific or degenerative arteriosclerotic aortic heart valve disease. PMID- 9740512 TI - Ulcer bed infection. Report of a case of enlarging venous leg ulcer colonized by Pseudomonas aeruginosa. AB - We report a case of ulcer bed infection in an enlarging venous leg ulcer without clinical signs of cellulitis in the surrounding tissues. Signs of infection in the leg ulcer were: 1) cocci-like structures and bacteria-like rods around vessel walls in the viable ulcer bed, 2) vasculitis-like inflammation of deeply situated vessels of the viable tissue, 3) Pseudomonas aeruginosa-specific antibodies in the serum (other than against exotoxin A), 4) extensive epidermolysis of normal human skin by the wound exudate in vitro, and 5) P. aeruginosa exotoxin A in the wound exudate (23 ng/ml). In an in vitro cell assay, the wound exudate was cytotoxic and rabbit antibodies to exotoxin A, but not a serine proteinase inhibitor, inhibited this cytotoxicity. P. aeruginosa exotoxin A might contribute to the pathogenesis of the ulcer enlargement. The ulcer improved after the third skin graft, probably mainly due to effective treatment with a long-stretch compression bandage. PMID- 9740513 TI - Reappraisal of the expression of mast cell proteases of Mongolian gerbils (Meriones unguiculatus). AB - Mast cell proteases in the tongue and jejunum of Mongolian gerbils (Meriones unguiculatus) were examined by enzyme-histochemical methods. Both trypsin-like (tryptase) and chymotrypsin-like (chymase) protease activities were demonstrated in mast cells in the tongue of fresh cryosections. When frozen sections of the tongue were post-fixed in various fixatives, those fixed in Carnoy's fluid showed strongest enzyme activities. Tryptase and chymase activities in paraffin sections of both tissues were well preserved when tissues were fixed in Carnoy's fluid at 4 degrees C for 15 min. However, enzyme activities in both tissues, especially in the tongue, were drastically reduced by longer fixation time and higher temperature. When Carnoy-fixed (4 degrees C for 15 min) paraffin sections were treated with heparinase I or chondroitinase ABC before enzyme-histochemical stainings for proteases, tryptase activities were lost entirely in the tongue and mostly in the jejunum by heparinase I digestion, and slightly in both organs by chondroitinase ABC digestion. In contrast, chymase activities at both sites were not influenced by these pretreatments. These results show that although mast cells in the tongue as well as in the jejunum of Mongolian gerbils contain both tryptase and chymase activities, their stability to fixations is variable among organs so that tissue fixation conditions are crucial for the preservation. At least some part of the stability of mast cell proteases is dependent on the proteoglycans present in mast cell granules. PMID- 9740514 TI - Adhesion to denture acrylic surfaces and relative cell-surface hydrophobicity of Candida parapsilosis and Candida albicans. AB - C. parapsilosis is an opportunistic emerging pathogen which together with C. albicans causes diseases in immunocompromised patients. Adhesion of Candida species to various surfaces is an important event in colonization and pathogenesis, and the relative cell-surface hydrophobicity (CSH) of the organism is a contributory physical force involved. Therefore, in vitro adhesion to acrylic surfaces and relative CSH of 24 isolates of C. parapsilosis and 10 isolates of C. albicans were studied. There was no significant difference in relative adhesion of C. parapsilosis isolates and C. albicans, although the former demonstrated a tendency for increased adhesion. There was significant intra-species variation in adhesion among isolates of C. parapsilosis (p=0.0001), but not C. albicans. In general, C. parapsilosis isolates demonstrated a two-fold greater relative CSH than C. albicans (p=0.0003). When the relative CSH of superficial and systemic isolates of C. parapsilosis were compared, the former showed a significantly higher (49.15%) relative CSH than their systemic counterparts (p<0.01). A highly significant positive correlation between adhesion and relative CSH of C. parapsilosis (p=0.74, p<0.0001) was also noted. Taken together, these data suggest that the attributes of adhesion and relative CSH of Candida species may contribute differentially in varying disease states of the human host, such as superficial and systemic Candida infections. PMID- 9740515 TI - IL-1beta, IL-6, TNFalpha, fetal fibronectin, and endotoxin in the lower genital tract of pregnant women with bacterial vaginosis. AB - BACKGROUND: In our studies on women with bacterial vaginosis (BV) in early pregnancy a strong association has been found between BV and the levels of endotoxin or interleukin-1alpha (IL-1alpha) in the lower genital tract. In the present study we investigated if an association could be found between BV and other cytokines (IL-1beta, IL-6, tumor necrosis factor alpha, TNF) or fetal fibronectin (FFN). The cytokine-inducing capacity of endotoxins present in the cervical mucus was explored in a monocytic cell assay. METHODS: Cervical mucus or cervicovaginal fluid was collected from women with (BV) and without BV (nonBV) attending a family planning unit for first trimester abortion. The concentrations of IL-1beta, IL-6, TNF and FFN were determined by quantitative enzyme immunoassays. TNF was determined in 63 women (BV, n=25) out of whom 37 (BV, n=11) were analyzed for IL-1beta and the remaining 26 for IL-6 (BV, n=14). FFN was determined in another 36 women (BV, n= 19). The cytokine-inducing capacity of endotoxin-containing cervical mucus and purified endotoxin of Prevotella bivia were studied by an in vitro cell assay using a human monocytic cell line (THP-1). RESULTS: IL-lbeta and IL-6 were found in almost all women. The levels of IL 1beta, but not IL-6, TNF or FFN, were significantly increased in women with BV compared with the nonBV women (p<0.05). Purified endotoxin from P. bivia, and cervical mucus from BV women containing high levels of endotoxin were able to induce a cytokine response (IL-6) in monocytic cells in vitro. CONCLUSION: BV is associated with increased levels of IL-1beta in the lower genital tract of pregnant women in the first trimester. The ability of BV-associated endotoxins to induce cytokine production in monocytic cells may partly explain the increased IL 1beta levels. PMID- 9740516 TI - Short-term maternal oxygen administration in fetuses with absence or reversal of end-diastolic velocity in umbilical artery: pathophysiological and clinical considerations. AB - BACKGROUND: The aim of our study was to evaluate the hemodynamic response to acute maternal hyperoxygenation (O2 test) in a group of growth retarded fetuses with absence or reversal of end-diastolic velocity (AREDV) in the umbilical artery (UA) and to correlate this response to a series of feto-placental velocimetric parameters and clinical variables. METHODS: In 25 singleton pregnancies, feto-maternal Doppler velocimetry was performed before and after acute maternal hyperoxygenation. RESULTS: Ten fetuses (40%) exhibited an increase of middle cerebral artery Pulsatility Index (PI) >20% after O2 (Responders), while in 15 fetuses PI did not change relevantly (Nonresponders). Non-responder fetuses showed a higher prevalence of reverse flow in umbilical artery (6/15 vs 0/10: p<0.03) and a slight, but not significant, higher percentage with reversed flow in inferior vena cava (% of A). Also the prevalence of a % of A greater than 95th confidence interval was higher in Non-responders (13/15 vs 4/10; p<0.04). Finally the Responder fetuses showed higher peak velocities in the cardiac outflows, even if the difference reached a statistical significance only for the pulmonary artery. The outcome of the two groups did not differ significantly. CONCLUSIONS: Our results seem to prove an ability of O2 test in selecting a group of AREDV fetuses characterized by a higher degree of hemodynamic deterioration and hence 'placed' in a more advanced step of the pathological process leading to overt cardiac decompensation, even if the clinical application of such a test seems to be still of limited value. PMID- 9740517 TI - Seasonal change in the incidence of preeclampsia in Zimbabwe. AB - BACKGROUND: The aim of our study was to evaluate the number of women with hypertensive complications during pregnancy in a southern province in Zimbabwe and to examine the annual change in the incidence of preeclampsia. METHODS: In three different hospitals the preeclamptic women who were treated between January 1992 and August 1995 were counted. This data was compared with the amount of rainfall obtained from the local meteorological stations. RESULTS: A distinctive change in the incidence of preeclampsia during the year could be observed. These changes go along with the seasonal variation in precipitation: at the end of the dry season and in the first months of the rainy season there is an increase in the incidence. CONCLUSIONS: The relationship between climate and occurrence of preeclampsia raises new questions in the pathophysiology of preeclampsia. Possible explanations could be the impact of humidity and temperature on vessels or the production of vasoactive substances. Dry and rainy seasons influence the agricultural yields and therefore the nutritional status could also play a role in the pathophysiology. PMID- 9740518 TI - Red cell age and susceptibility to malaria during pregnancy. AB - BACKGROUND: Increased susceptibility to malaria in pregnancy is well recognized, and has generally been assumed to be due to hormonal changes resulting in altered immunity. Based on previous work demonstrating enhanced parasite growth in young normal and thalassemic red blood cells, we hypothesized that in pregnancy increased malaria susceptibility may be due, in part, to the increase in the population of young red cells. METHODS: FC27 strain of Plasmodium falciparum was cultured in the red cells and sera from healthy primigravida pregnant (n=9) and non-pregnant (n=9) women. Red cells from both pregnant and non-pregnant women were each placed in three cultures containing the sera from pregnant, non pregnant and pooled control samples. Cultures were set up in triplicate and incubated for 144 hours. Parasite development and growth were assessed by slide microscopy. RESULTS: At 96 hours the median parasite growth in cells from pregnant samples (5.7%) was significantly better than that in the non-pregnant cells (4.8%) (p=0.01). There was no significant difference in parasite growth in cultures with pregnant and non-pregnant sera. As expected, there were significant differences in parameters measuring red cell age between the cells from pregnant and nonpregnant samples: median red cell creatine (11.09 mg/dl) versus (6.90 mg/dl) (p=0.004) and median reticulocyte count (2.3%) versus (1.4%) (p=0.0006). CONCLUSIONS: These preliminary results are consistent with the hypothesis that an increased population of young red cells may contribute to increased malaria susceptibility during pregnancy. PMID- 9740519 TI - Clinical and pathologic correlates of stillbirths in a single institution. AB - BACKGROUND: To evaluate risk factors, placental and pathologic determinants of stillbirths. METHODS: A retrospective analysis of stillbirths > or = 25 weeks was performed. Clinical data was compared to a randomized control group. Statistical analysis included chi square test, student t test, and logistic regression. RESULTS: One hundred and fifteen stillbirths and 193 controls were analyzed. Maternal age, nulliparity, tobacco use, previous induced abortions, anticardiolipid antibodies, elevated maternal serum alpha feto protein, twins, and amniocentesis, were significantly associated with stillbirth. Logistic regression analysis showed only maternal age, tobacco use, small for gestational age (SGA), previous induced abortions, decreasing gestational age as independent significant variables. The stillbirth baby was 6.8 times more likely to be SGA and 11.9 times more likely to be preterm. Primary pathologic diagnoses were placental factors (37%), cord complications (28%), and fetal causes (15%), 17% had maternal risk factors only and 3% had no known risk factors. Diagnosis was suggested by pathology in 40% of cases. CONCLUSIONS: Stillbirth delivery is associated with older, nulliparous patients with prenatal complications resulting in intrauterine growth retardation and prematurity. Perinatal histopathologic examination is important in diagnosis. Utilizing an extensive testing protocol will reduce the diagnosis of unexplained stillbirth. PMID- 9740520 TI - Interest in alternative birth settings in Finland. AB - BACKGROUND: The aim of this study was to examine the extent of wishes for and realised choices of alternative and conventional birth care in Finland based on survey and registry data. METHODS: A population-based national survey to 3000 women of reproductive age and 400 men aged 18-24 and 40-44 in Finland and a cross sectional analysis of all childbirths in 1990 1995 based on the National Medical Birth Registry (n=390, 943). RESULTS: In the survey 69% of women and 66% of men chose conventional hospital birth as their preferred alternative. An early discharge birth was chosen by 14% of women and 18% of men and home birth by 6% of women and 3% of men. Childless respondents were more likely than parents to choose an alternative other than conventional hospital birth, yet 16% of mothers and 14% of fathers would choose either home birth or early discharge from hospital in a future birth. The expressed interest in alternatives to conventional hospital care was far greater than what occurs in reality: in the MBR data 99% of births were conventional hospital births, 0.01% were planned homebirths and 0.9 % early discharge births. CONCLUSIONS: The study shows a discrepancy between expressed interests and actually realised choices of birth settings. The majority of female and male survey respondents would choose conventional hospital care for birth. However, the fact that even some women who had earlier birth experience preferred some form of alternative to the conventional hospital birth should be taken as a sign of women wanting choices in birth care. PMID- 9740521 TI - Third degree obstetric tears; outcome after primary repair. AB - BACKGROUND: Disruption of the anal sphincter occurs in 0.5 to 2.5% of women during delivery. Defects of the sphincter are major causes of fecal incontinence. More than 30% of women who suffer from third degree perineal tears develop incontinence. We sought to determine the incidence of symptoms and injury to the anal sphincter among women who gave birth during a 5 year period. We also investigated the sensitivity of manometry and endosonography as well as the correlation of these two diagnostic modalities. METHODS: Thirty-eight women were examined one to five years after delivery. We used a questionnaire to assess symptoms of anal incontinence. Anal manometry and endosonography were performed. RESULTS: Twenty (57%) women had symptoms; most of them (34%) in the form of flatulence incontinence. The rest were incontinent of either liquid or solid stools. Four of these women were re-operated. Seventeen percent of the women suffered from anal incontinence during sexual intercourse. Only seven women had been in contact with a doctor regarding these problems. CONCLUSION: The fact that 57% of the women that took part in this study reported complications, leads us to the conclusion that the primary repair of third degree anal sphincter tears is unsatisfactory. It is important to decide whether any changes in primary repair may improve results in the future. Sexual dysfunction is also a complication of third degree obstetric tear with primary repair. It is important that the women who suffer from anal sphincter tear, as well as doctors, are given information about possible symptoms and the treatment available. PMID- 9740522 TI - Closure versus non-closure of peritoneum at cesarean section--evaluation of pain. A randomized study. AB - OBJECTIVE: To evaluate the effects of leaving the parietal peritoneum open at lower segment cesarean section (LSCS) measured by postoperative pain. DESIGN: A randomized, prospective and double-blind study. SETTING: Department of Obstetrics and Gynecology, Aarhus University Hospital, Denmark. POPULATION: Forty women referred for an elective cesarean section were assigned to one of two groups: peritoneum open (n=21) or peritoneum closed (n=19). METHODS: Pain was evaluated twice a day from the first to the fifth postoperative day by Visual Analog Scales. MAIN OUTCOME MEASURES: Postoperative pain. Other outcomes include usage of analgesics, bowel function, postoperative complications, and hospital stay. RESULTS: We found no overall difference in postoperative pain. A tendency to less pain was found in the non-closure group from the third postoperative day to the fifth postoperative day. No differences were found either in the incidence of postoperative complications, or the time to return of bowel function. Concerning opiate analgesics the non-closure group had a significantly higher use in the second postoperative 24-hour period, but in the remains of the registration period it was significantly lower. For oral analgesics no difference was found in the first 24-hour period, but in the remains of the period the non-closure group had a significantly lower use. CONCLUSIONS: The VAS-scales showed no difference in postoperative pain comparing closure to non-closure of the parietal peritoneum. However, the use of analgesics is lower in the non-closure group. We suggest leaving the parietal peritoneum open when performing LSCS. PMID- 9740523 TI - Fertility and long-term complications four to nine years after appendectomy during pregnancy. AB - BACKGROUND: The purpose of this study was to determine the long-term outcome after an appendectomy during pregnancy, especially focusing on fertility. METHODS: The Danish National Registry of Patients identified 117 pregnant women who had had an appendectomy during the period 1980 to 1985. One hundred-and-one of these women answered a questionnaire designed to focus on long-term complications, including infertility, during a 4 to 9 year period after the appendectomy. More than a 2-year attempt to conceive was defined as infertility. RESULTS: Of the 101 women 15 with a normal appendix had a new intraperitoneal operation due to different indications. Three of these women had intraperitoneal adhesions. In one patient, adhesions were located on the Fallopian Tube but the location did not influence fertility. Five of the 101 women complained of infertility as defined; all had a normal appendix and none had intraperitoneal surgery during the observation period. Two of these five women conceived later during the observation period; one had confounding female and male infertility factors and two were not examined. CONCLUSIONS: Appendectomy during pregnancy of a normal, inflamed or perforated appendix does not seem to cause clinically significant intraperitoneal adhesions or infertility later in life. PMID- 9740524 TI - Factors influencing the presence of uterine cavity fluid in a random sample of asymptomatic postmenopausal women. AB - AIMS: To assess possible endometrial pathology and other factors influencing the presence of uterine cavity fluid in postmenopausal women. STUDY DESIGN: A random sample of 559 asymptomatic postmenopausal women, recruited from the total population, were examined by transvaginal sonography (TVS) for the presence of uterine cavity fluid. Women with uterine cavity fluid who had an endometrial thickness of > or = 8 mm (including fluid) were admitted for hysteroscopy and a dilatation and curettage (D & C), and those with <8 mm underwent a new TVS examination one year later. A medical history, including details regarding previous minor gynecological surgery, was taken from the women and from an age matched control-group of women from the same population. RESULTS: Uterine cavity fluid was found in 8.9% (50/559) of the women. In four women with an endometrium measuring > or = 8 mm, curettage revealed polyps in three women and atrophy with a pyometra in one woman. At the one-year follow-up, 22 women who originally had an endometrial thickness<8 mm had an endometrial thickness of<5 mm; 11 women had no cavity fluid and in the remaining 11 the cavity fluid had decreased. In 17 women, endometrial thickness measured > or = 5 mm and subsequent histology showed 11 endometrial biopsies with atrophy, four endometrial polyps and two cervical polyps. The prevalence of uterine cavity fluid increased with increasing age (p<0.0001) and was increased in smokers (p<0.013) but was unaltered by the presence or absence of hormone replacement therapy (HRT). CONCLUSION: There were no indications that uterine cavity fluid was associated with malignancy. The prevalence of uterine cavity fluid increased with increasing age and was higher in smokers. We could not demonstrate an increased prevalence of fluid in HRT users. PMID- 9740525 TI - Endometrial morphology during hormone replacement therapy with estradiol gel combined to levonorgestrel-releasing intrauterine device or natural progesterone. AB - OBJECTIVES: To evaluate endometrial responses to three different forms of amenorrhea-inducing HRT in postmenopausal women. MATERIAL AND METHODS: Fifty-one postmenopausal women completing a one-year HRT trial with percutaneous estradiol gel containing 1.5 mg estradiol daily combined with a levonorgestrel-releasing intrauterine device (LNG-IUD) (n=18), or natural progesterone 100 mg daily orally (n= 19) or vaginally (n=15) during 1-25 calendar days of each month. Endometrial thickness and uterine size were measured by transvaginal ultrasound, and endometrial cytology/histology was assessed from specimens taken by needle aspiration before the study and at 12 months. RESULTS: Before medication, the median endometrial thickness was 2.0 mm in the LNG-IUD group, 2.4 mm in the oral P group and 2.5 mm in the vaginal P group. At 12 months of therapy the respective values, 3.0, 2.7 and 2.4 mm, did not differ significantly from the initial values. LNG-IUD induced epithelial atrophy in all women, which was accompanied by stromal decidualization in 12 women. On the contrary, only four women in the oral P group and five women in the vaginal P group had an inactive or atrophic endometrium. The remaining cases were dominated by proliferative features. No hyperplasia was seen in any of the groups. CONCLUSION: LNG-IUD appeared to be an effective method of counteracting the stimulatory effect of estrogen on the endometrium, whereas natural progesterone given orally or vaginally was not sufficiently effective in this function at the doses used. The vaginal and oral administrations of progesterone did not differ from each other in this respect. PMID- 9740526 TI - A multicenter study comparing two endometrial sampling devices--Medscand Endorette and Pipelle de Cornier. AB - BACKGROUND: To compare two endometrial sampling devices Medscand Endorette and Pipelle de Cornier with respect to tissue collecting ability, diagnostic accuracy and side effects. METHODS: A prospective, multi-center, cross-over study in 152 women with a medical indication for endometrial biopsy. Samples were collected from each patient on the same occasion with both devices, the order of which was randomized. Statistical analysis was based on pairwise comparison in contingency tables with McNemars chi2 test. RESULTS: One hundred and forty-five of 152 (95%) women were successfully sampled. There was no difference between the devices concerning discomfort and bleeding. However, Medscand Endorette seemed to have a higher capacity than Pipelle for collecting an adequate sample. This difference was noted when the two devices were compared in the position as first instrument as well as when results were compared in the main target group for this type of examination, for women over 55 years. CONCLUSIONS: The new Medscand Endorette was preferred due to its higher capacity for collecting adequate samples. PMID- 9740527 TI - Hysterectomy trends in Finland in 1987-1995--a register based analysis. AB - BACKGROUND: The study objective was to identify trends in the use of hysterectomy by nationwide register based analysis in Finland. METHODS: All women (n=89,069) undergoing hysterectomy in 1987-1995 according to the Finnish Hospital Discharge Register were the numerator. The annual denominator data were obtained from the population database of Statistics Finland. RESULTS: From 1987 to 1992 the hysterectomy rate increased by 22%, from 340 to 414 per 100,000 females, almost half of this being attributable to the changing age structure. From 1993 on, ambiguity in coding laparoscopically assisted vaginal hysterectomies prohibited detailed analyses. However, the overall trend continued at least among women 50 years and over until 1995. The age-adjusted 12% increase from 1987 to 1992 coincided with a rapid increase in operation rates in postmenopausal groups (60% or more among women aged 55 59 and 70-79 years). Among women aged 55-64 years, operations for fibroids and uterine bleeding more than doubled, suggesting an influence of increased use of estrogen replacement therapy. Among all women, operations due to bleeding disorders and genital prolapse showed the largest increase (41% and 42% respectively). Bilateral oophorectomy became more common in all age groups over 46 years. CONCLUSIONS: There was a modest increase in the overall hysterectomy rate. However, the operation became far more common in postmenopausal women, possibly due to the growing use of estrogen replacement therapy. Register data can be used for describing changes in clinical practice, but other methods are needed to confirm the causal relationships underlying the changes. PMID- 9740528 TI - Prognosis of 2,800 patients with epithelial ovarian cancer diagnosed during 1975 94 and treated at the Norwegian Radium Hospital. AB - BACKGROUND: Ovarian cancer patients have a poor prognosis. In Norway, however, the prognosis has improved steadily since the 1950s, the age-adjusted 5-year relative survival reaching 37% in 1989 93. The aim of the present study was to explore the prognosis of patients with epithelial ovarian cancer diagnosed during 1975-94 (the prepaclitaxel period) and treated at The Norwegian Radium Hospital. METHOD: Relative risks (RR) of dying and 95% confidence intervals (95% CI) were derived from multivariate Cox proportional hazards regression models. RESULTS: A total of 2,769 patients with epithelial ovarian cancer were included in the present study. Altogether 54% of the patients were diagnosed with advanced stage disease (stages III and IV), whereas 32% were diagnosed with stage I disease. The prognosis of the patients improved from the 1970s to the 1990s, mainly due to increased short-term survival. In multivariate survival analysis, the RR of dying decreased with period of diagnosis. An RR of 0.77 (95% CI=0.66-0.89) was seen in 1990-94 compared with 1975-79. CONCLUSION: The short-term survival of patients with epithelial ovarian cancer improved from the late 1970s to the early 1990s. However, no major improvement in the long-term survival was seen. PMID- 9740529 TI - Morbidity after cesarean section in obese women. PMID- 9740530 TI - Transvaginal ultrasound detection in early pregnancy of cystic hygroma associated with fetal chromosomopathies. PMID- 9740531 TI - B 19 parvovirus infection in a primipara with congenital spherocytosis. PMID- 9740532 TI - Maternal supraventricular tachycardia recorded as apparent fetal heart rate in a case of fetal demise. PMID- 9740533 TI - Emergency combined cesarean section and thoracoscopic pleurodesis in a patient with recurrent spontaneous pneumothorax. PMID- 9740534 TI - Uterine inversion during cesarean section. PMID- 9740535 TI - The role of lipid-lowering therapy in multiple risk factor management. AB - Epidemiological studies have demonstrated that the risk of death from coronary heart disease (CHD) increases in parallel with increasing serum cholesterol levels. The risk is highest in patients with severely elevated cholesterol levels [i.e. > 6.2 mmol/L(240 mg/dl)]. However, even in patients with serum cholesterol levels of 4.7 to 5.7 mmol/L (180 to 220 mg/dl), mortality from CHD is 30 to 70% higher than in patients with cholesterol levels < 4.7 mmol/L (180 mg/dl). A number of factors other than serum total cholesterol levels affect CHD risk. It is increasingly accepted that dysfunction of the vascular endothelium contributes to the pathogenesis of atherosclerosis. Experimental evidence and clinical studies suggest that endothelium-derived nitric oxide (NO) plays an important role as an endogenous antiatherogenic molecule, and that reduced levels of NO may promote progression of atherosclerosis. Hypercholesterolaemia can hasten atherogenesis in part by reducing levels of NO. In humans, it is possible to evaluate the effects of hypercholesterolaemia by measuring the vasodilator response to pharmacological or physical stimuli that increase the synthesis and release of endothelium-derived NO. The use of high resolution external ultrasound to assess post-ischaemic brachial artery vasodilation in patients with cardiovascular risk factors has demonstrated that endothelial dysfunction exists, even in the absence of overt atherosclerotic plaques. Impaired endothelium dependent vasodilation has been observed in asymptomatic individuals with hypercholesterolaemia, as well as in patients with other recognised risk factors. Dietary and pharmacological lipid lowering has been shown to produce improvements in endothelial-dependent vasodilation. Among normocholesterolaemic healthy young individuals without cardiac risk factors, endothelium-dependent vasodilation was more likely to be impaired in those with relatively higher total and low density lipoprotein-cholesterol levels (i.e. above versus below the 25th percentile). This suggests that an inverse and continuous relationship exists between the prevailing cholesterol level and endothelial function. The implication of these findings is that the treatment of hypercholesterolaemia remains, at present, an underutilised means of reducing CHD. PMID- 9740536 TI - New insights into plaque stabilisation by lipid lowering. AB - Thrombosis on the substrate of a disrupted plaque causes most acute coronary events. The physical integrity of the plaque thus governs the most important clinical manifestations of atherosclerosis. Of particular importance is the extracellular matrix of the fibrous capsule overlying the thrombogenic core of the atheroma. Stable atheroma generally have thick fibrous caps, and smaller lipid cores than lesions that have ruptured. Accumulating evidence supports a key role for inflammation as another critical determinant of the stability of human atherosclerotic plaques. Plaques that rupture usually have more abundant leucocytic infiltrates than those considered stable. Inflammatory mediators such as cytokines can influence several biological processes that regulate the stability of the plaque's fibrous cap, and thus its resistance to rupture. For example, interferon-gamma produced by activated T lymphocytes within atheroma inhibits the production of interstitial forms of collagen by human vascular smooth muscle cells. Inflammatory cytokines such as interleukin-1, tumour necrosis factor (TNF) and CD-40 ligand (a cell surface homologue of TNFalpha) can also elicit the expression by macrophages and smooth muscle cells of proteolytic enzymes that can weaken the extracellular matrix. We have hypothesised that lipid lowering reduces stimuli for the inflammatory response within the complex atherosclerotic lesion. Recent studies in rabbits with experimentally produced atherosclerosis have indeed shown that lipid lowering can (i) reduce macrophage numbers, (ii) decrease expression of the collagenolytic enzyme MMP-1, and (iii) reinforce the plaque's fibrous skeleton by increasing the content of interstitial collagen. By reducing local inflammation, lipid lowering can thus stabilise the plaque's fibrous cap, rendering the atheroma less prone to rupture and to precipitate thrombotic complications. These observations provide a mechanistic basis for understanding the marked reduction in acute coronary events and cerebrovascular accidents observed in patients treated with agents that reduce plasma lipids. PMID- 9740537 TI - Rational assessment of the interaction profile of cerivastatin supports its low propensity for drug interactions. AB - Pharmacokinetic drug-drug interactions influence drug efficacy, tolerability, and compliance. Such interactions are both more common and of more clinical relevance than often appreciated. The US Food and Drug Administration and the European Agency for the Evaluation of Medicinal Products have recently issued guidelines setting out in vitro and in vivo investigations to be conducted during drug development. These guidelines reflect the increasing interest of public health authorities in this topic. Cerivastatin is a novel, potent HMG-CoA reductase inhibitor that effectively reduces serum cholesterol levels at low daily doses. It is completely absorbed after oral administration, undergoes moderate first pass metabolism and high plasma protein binding, and is cleared exclusively via hepatic cytochrome P450 (CYP). Unlike other drugs of its class, cerivastatin has a dual metabolic pathway, with the involvement of more than one CYP isozyme. Metabolites are cleared via both biliary and renal excretion. On the basis of this pharmacokinetic profile and a knowledge of the target population, the formal in vivo interaction programme for cerivastatin investigated many important potential cerivastatin drug-drug interactions. Cerivastatin appears to lack clinically relevant interactions with digoxin, warfarin, antacid, cimetidine, nifedipine, omeprazole, erythromycin and itraconazole. PMID- 9740538 TI - Extending therapy options in treating lipid disorders: a clinical review of cerivastatin, a novel HMG-CoA reductase inhibitor. AB - Cerivastatin is a third generation pure enantiomeric HMG-CoA reductase inhibitor. It reduces low density lipoprotein (LDL)-cholesterol by 22 to 44% at doses of 0.1 to 0.8 mg/day. The drug has been extensively evaluated for more than 5 years in clinical trials and is currently marketed in a number of countries at doses of 0.1 to 0.3 mg/day. Cerivastatin has been tested in more than 4000 patients during extensive phase II and III studies. About 40% of patients in these trials were women, and many participants were aged between 65 and 75 years. The trial populations had moderate to severe hypercholesterolaemia, with mean baseline LDL cholesterol levels of approximately 5.2 mmol/L (200 mg/dl). In large phase III trials, cerivastatin, over the dosage range of 0.1 to 0.4 mg/day, reduced LDL cholesterol by 22.4 to 36.1% from baseline. As with other HMG-CoA reductase inhibitors, the log-linear dose-response curve of cerivastatin showed a 6% additional decrease in mean LDL-cholesterol levels for each doubling of the daily dose, with no plateau effect noted at the highest dosage yet tested (0.8 mg/day). High density lipoprotein cholesterol levels increased by 4 to 10% during cerivastatin therapy. This effect, which was consistent with that of other HMG CoA reductase inhibitors, was not dose related. As has been found with other statins, the triglyceride-lowering effects of cerivastatin are dependent on baseline triglyceride levels, with very small reductions occurring in patients with low initial levels [< 1.7 mmol/L (150 mg/dl)], and larger dose-dependent reductions of up to 36% with the 0.4 mg/day dose observed in patients with baseline triglyceride levels >2.8 mmol/L (250 mg/dl). Cerivastatin was well tolerated in all studies. Cerivastatin recipients and recipients of other HMG-CoA reductase inhibitors experienced a similar incidence of adverse events (including hepatic transaminase elevations) in comparative studies. Cerivastatin is an effective and safe lipid-lowering agent for most patients with hypercholesterolaemia. PMID- 9740539 TI - Vinflunine (20',20'-difluoro-3',4'-dihydrovinorelbine), a novel Vinca alkaloid, which participates in P-glycoprotein (Pgp)-mediated multidrug resistance in vivo and in vitro. AB - Vinflunine (VFL) is a novel derivative of vinorelbine (NVB, Navelbine), which has shown markedly superior antitumor activity to NVB, in various experimental animal models. To establish whether this new Vinca alkaloid participates in P glycoprotein (Pgp)-mediated multidrug resistance (MDR), VFL-resistant murine P388 cells (P388/VFL) were established in vivo and used in conjunction with the well established MDR P388/ADR subline, to define the in vivo resistance profile for VFL. P388/VFL cells proved cross-resistant to drugs implicated in MDR (other Vinca alkaloids, doxorubicin, etoposide), but not to campothecin or cisplatin and showed an increased expression of Pgp, without any detectable alterations in topoisomerase II or in glutathione metabolism. The P388/ADR cells proved cross resistant to VFL both in vivo and in vitro, and this VFL resistance was efficiently modulated by verapamil in vitro. Cellular transport experiments with tritiated-VFL revealed differential uptake by P388 sensitive and P388/ADR resistant cells, comparable with data obtained using tritiated-NVB. In various in vitro models of human MDR tumor cells, whilst full sensitivity was retained in cells expressing alternative non-Pgp-mediated MDR mechanisms, cross resistance was identified in Pgp-overexpressing cells. Differences were, however, noted in terms of the drug resistance profiles relative to the other Vinca, with tumor cell lines proving generally least cross-resistant to VFL. Overall, these results suggest that VFL, like other Vinca alkaloids, participates in Pgp-mediated MDR, with tumor cells selected for resistance to VFL overexpressing Pgp, yet MDR tumor cell lines proved generally less cross resistant to VFL relative to the other Vinca alkaloids. PMID- 9740540 TI - Pharmacokinetic and phase I studies of brequinar (DUP 785; NSC 368390) in combination with cisplatin in patients with advanced malignancies. AB - Brequinar (DUP 785; NSC 368390) is a quinoline carboxylic acid derivative that inhibits pyrimidine synthesis at the level of dihydro-orotate dehydrogenase and revealed synergy with cisplatin in preclinical models. In this study investigating the pharmacokinetic and toxicity of brequinar in combination with cisplatin, patients were initially treated with weekly brequinar, in combination with an every-three-week administration of cisplatin. Due to toxicity, the schedule was modified to a 28-day cycle with brequinar given on days 1, 8, 15, and cisplatin on day 1. A total of 24 patients (16 male, 8 female; median age 57; median performance status 1) received 69 courses of therapy. Six dose levels were explored, with cisplatin/ brequinar doses, respectively, of 50/500, 50/650, 50/860, 60/860, 75/650, and 75/860 mg/m2. The serum concentration versus time curves for brequinar were biphasic. A comparison of the pharmacokinetic results after the first and third doses of brequinar indicate that the presence of 50, 60, and 75 mg/m2cisplatin did not change the protein binding and the pharmacokinetics of brequinar in any of the three brequinar-dose groups. Total cisplatin plasma pharmacokinetic followed a triphasic-shape curve and unbound cisplatin decayed at a very rapid rate. Since pharmacokinetic parameters for total cisplatin in this study were similar to those reported in the literature, the presence of brequinar is unlikely to alter the pharmacokinetics of cisplatin. Main dose-limiting toxicities included myelosuppression (including neutropenia and thrombocytopenia) and mucositis. Cisplatin/brequinar doses of 50/500, 50/650, 50/860, 60/860, 75/650, and 75/860 mg/m2, were associated with dose limiting toxicity in 0/3, 1/3, 1/3, 1/3, 2/4, 2/5, and 4/6 patients, respectively. This study shows that co-administration of brequinar and cisplatin does not affect the pharmacokinetic properties of either drug and that the MTDs of cisplatin/brequinar combinations are 60/860 mg/m2 or 75/650 mg/m2. From this study, we conclude that full dose of 75 mg/m2 cisplatin (day 1) can be administered with 650 mg/m2 brequinar (days 1, 8 and 15) without significant modifications of individual drug pharmacokinetic parameters. PMID- 9740541 TI - Phase I study of paclitaxel (taxol) and granulocyte colony stimulating factor (G CSF) in patients with unresectable malignancy. AB - A phase I trial of a 24-hour infusion of paclitaxel was conducted to identify the maximum tolerated dose of paclitaxel with granulocyte colony-stimulating factor (G-CSF) in patients with unresectable malignancy previously untreated with chemotherapy. Nineteen patients with metastatic melanoma or non-small cell lung cancer were treated with paclitaxel administered at 250, 300, 400 mg/m2 every 3 weeks. G-CSF, 5 microg/kg was given as a daily subcutaneous injection 24 hours after the completion of the infusion. Dose limiting myelosuppression and peripheral neuropathy was observed at 400 mg/m2 and 350 mg/m2. Paclitaxel can be safely administered as a 24-hour infusion at 300 mg/m2 with G-CSF. Further studies of paclitaxel and G-CSF are recommended to determine a dose-response relationship in sensitive tumors. PMID- 9740542 TI - A phase I trial of vinorelbine in combination with mitoxantrone in patients with refractory solid tumors. AB - Vinorelbine (Navelbine) is a unique semi-synthetic vinca-alkaloid with a favorable safety profile that has demonstrated significant antitumor activity in patients with non-small cell lung cancer, advanced breast cancer, advanced ovarian cancer and Hodgkin's disease. The most common dose-limiting toxicity is neutropenia, while other reported toxicities are minimal. Mitoxantrone (Novantrone) is an anthracene derivative that has demonstrated antitumor activity in patients with breast cancer, ovarian cancer, acute leukemia, and lymphoma. Mitoxantrone also has a very favorable toxicity profile with significantly less nausea and vomiting, alopecia, and stomatitis as compared with anthracyclines. The dose-limiting toxicity for mitoxantrone is leukopenia. The study was designed to determine the safety and maximally tolerated dose of IV vinorelbine used in combination with a fixed dose of mitoxantrone for the treatment of patients with refractory solid tumors. Vinorelbine was administered on days 1 and 8 of the treatment regimen as a short IV infusion. The starting dose was 15 mg/m2. Mitoxantrone was administered as a 20-min infusion on day 1 only at a fixed dose of 10 mg/m2. Seventeen patients with solid malignancies were entered in the study. For personal reasons, one patient decided to discontinue the treatment after day 1 of cycle 1. Therefore, 16 patients were evaluable for toxicity. The main toxicity was myelosuppression which was dose-limiting and resulted in dose reductions and delays. The use of G-CSF had a minimal overall impact on this regimen. Stable disease was observed in three cases. In patients previously treated with chemotherapy, the maximally tolerated dose was defined as vinorelbine 20 mg/m2 on days 1 and 8 and mitoxantrone 10 mg/m2 on day 1 without growth factor support. These doses can be recommended for phase II study of the regimen as salvage treatment. PMID- 9740543 TI - Phase I study of DMP 840 in pediatric patients with refractory solid tumors. AB - The bis-naphthalimide DMP 840 has demonstrated high level antitumor activity in a number of preclinical models and has been evaluated in several Phase I studies in adults. We enrolled 10 patients with refractory pediatric solid tumors to this Phase I study of DMP 840 given intravenously by short infusion daily for 5 days. The most frequent and dose-limiting toxicity was myelosuppression. The maximum tolerated dose on this schedule was 8.6 mg/m2 daily for 5 days. One patient had a complete response; there were no measurable tumor responses among the remaining 9 patients. PMID- 9740544 TI - Phase I clinical study of didemnin B. A National Cancer Institute of Canada Clinical Trials Group study. AB - Didemnin B (NSC-325319), a new depsipeptide isolated from a Caribbean tunicate, has been evaluated in a clinical phase I study. The drug was administered in a schedule of a 4 weekly intravenous injection in a six-weeks cycle. Fifty-three patients received 71 evaluable cycles in an escalated dose ranging from 0.4 mg/m2/week to 2.5 mg/m2/week. No hematological toxicity was demonstrated at any dose level. Without prophylactic antiemetics nausea and vomiting was dose limiting at 1.2 mg/m2/week. Due to the use of Cremophor EL as a solvent, hypersensitivity reactions occurred in 9 patients. These reactions occurred following prior exposure to the drug and were commonly seen at the 3rd dose. They were not dose related but became more frequent at 1.5 mg/m2/week necessitating prophylactic treatment with H and H2 receptor blocking agents. Non-hematological toxicities included mild diarrhea, mucositis, anorexia, headaches, and local phlebitis. The dose- limiting toxicity was generalized weakness which became severe and disabling in 3 of 6 patients treated at 2.5 mg/m2/week. No objective responses were documented in 39 patients with evaluable disease. The recommended dose for phase II studies was 2.3 mg/m2/week x 4 in a 6-weeks cycle given with prophylactic antiemetics and H1 and H2 receptor blocking agents. PMID- 9740545 TI - A phase I and pharmacokinetic trial of terephthalamidine (NSC 57155) as a 120 hour continuous infusion. AB - In this phase I study, terephthalamidine was administered as a 120-hour continuous infusion repeated every 21 days. Thirteen patients received 27 courses of terephthalamidine at four dose levels ( 14, 28, 46, and 70 mg/m2/day). Dose limiting toxicity consisted of profound and intractable anorexia, weight loss and prostration in all patients. Toxicity was delayed and accompanied by hyponatremia and hypokalemia. No hematologic or other toxicity was documented. One patient with adenocarcinoma of the lung had a 40% decrease in mediastinal lymph nodes and resolution of a pleural effusion lasting 2 months. Pharmacokinetic analysis by HPLC was performed in all patients during their first course. The harmonic mean terminal half-life for terephthalamidine was 23 hours with a plasma clearance of 1.7 1/hr/m2. Both plasma concentrations achieved during infusion (r2 = 0.9) and area under the curve (AUC) (r2 = 0.8) were proportional to increase in dose (p < 0.002). Renal excretion accounted for 64% of the total cumulative dose, with an average renal clearance of 1.16 1/hr/m2. Due to the unacceptable toxicity seen at all doses with this schedule, no further studies are recommended unless the mechanism of toxicity is better understood and can be prevented. PMID- 9740546 TI - Phase I trial of edatrexate plus carboplatin in advanced solid tumors: amelioration of dose-limiting mucositis by ice chip cryotherapy. AB - PURPOSE: Edatrexate (10-Edam) is a methotrexate analog with improved intracellular transport, polyglutamation, and antitumor activity compared to the parent compound. Edatrexate shows schedule-dependent synergism with platinum compounds in preclinical studies. We performed a phase I trial to determine toxicities and establish the maximum tolerated dose (MTD) of edatrexate in combination with carboplatin. Based on the short initial plasma half-life of edatrexate, prophylactic ice chip cryotherapy was used to reduce the severity of mucositis. PATIENTS AND METHODS: Forty-six chemotherapy-naive patients with advanced solid tumors were treated. Edatrexate was given weekly for 5 doses (50% on day 8), and then every other week, followed by carboplatin at a fixed dose of 350 mg/m2 on day 1 and then every 4 weeks for 8 cycles. Edatrexate dose was increased at increments of 10 mg/m2/dose level beginning at 60 mg/m2/week (range 60-120 mg/m2). RESULTS: All patients were assessable for toxicity and response analysis. The median number of cycles administered per patients was 4. This combination chemotherapy regimen was well tolerated. Using ice chip cryotherapy, no grade IV mucositis was observed. Grade III mucositis occurred in only 7/46 pts and was not dose-related. Protocol-defined dose-limiting toxicity occurred at a edatrexate dose level of 120 mg/m2, yielding an MTD of 110 mg/m2. Responding tumor types included non-small cell and small lung cancer, head and neck cancer, and bladder cancer. CONCLUSIONS: 1 ) This phase I study demonstrated the safety and tolerability of this edatrexate and carboplatin combination. 2) Dose-limiting mucositis did not occur allowing escalation of edatrexate dose above levels previously achieved with this edatrexate dose schedule. This was most likely a result of prophylactic ice chip cryotherapy. 3) An edatrexate dose of 110 mg/m2 with ice chip cryotherapy is recommended for Phase II trials of this combination. PMID- 9740547 TI - A phase II study of temozolomide in advanced untreated pancreatic cancer. AB - Temozolomide (SCH 52365) is an imidazotetrazine derivative which exhibits broad spectrum activity against murine tumors and is structurally related to dacarbazine (DTIC). Temozolomide cytotoxicity is schedule dependent in vivo with a daily x 5 schedule showing the highest activity. Oral temozolomide is rapidly and completely absorbed with minimal interpatient and intrapatient variability in pharmacokinetics. Clinical studies have demonstrated activity against melanoma and glioma. The present study examined the activity of oral temozolomide against patients with pancreatic cancer. Patients with advanced pancreatic adenocarcinoma previously untreated with chemotherapy received temozolomide 200 mg/m2/day once daily orally for 5 days with cycles repeated every 28 days. There were 16 patients entered on study with 15 evaluable for response and toxicity. There were no responses seen in 15 evaluable patients with 14 manifesting progressive disease within 2 months of starting therapy. Toxicity was primarily hematological with 3 patients experiencing > or = grade 3 neutropenia and thrombocytopenia respectively. Other toxicities were relatively modest. In conclusion, temozolomide in the once daily x 5 schedule is inactive against adenocarcinoma of the pancreas. PMID- 9740548 TI - Phase II study of intravenous adenosine 5'-triphosphate in patients with previously untreated stage IIIB and stage IV non-small cell lung cancer. AB - Fifteen patients with Stage IIIB or IV non-small cell lung cancer gave informed consent to receive three or more 96-hour infusions of ATP at a dose of 50 mcg/kg/min or higher to determine whether ATP has antineoplastic activity against this tumor type and to better define the spectrum of toxicity for ATP given as a single agent. There were no objective complete or partial responses observed. The median survival of the overall group was 187 days and the median time to tumor progression was 113 days. The major toxic side effects were chest pain and dyspnea, leading to the cessation of treatment in 5 patients. We conclude that ATP at this dose and schedule of administration is an inactive agent in patients with advanced non-small cell lung cancer. PMID- 9740549 TI - Phase II study of CI-980 (NSC 635370) in patients with previously treated advanced soft-tissue sarcomas. AB - Doxorubicin and ifosfamide are the two most active agents in the treatment of soft-tissue sarcomas. Patients whose tumors have failed these two drugs have very limited systemic therapy options. It is, therefore, important to identify newer drugs with activity against this disease. CI-980 is a synthetic mitotic inhibitor that binds to tubulin at the colchicine binding site and inhibits the polymerization of tubulin and blocks cell cycle progression in mitosis. Given its broad spectrum activity against several solid tumor models in vivo, we decided to perform a phase 2 study of this drug in previously treated soft-tissue sarcomas. A total of 18 eligible and evaluable patients were entered in the first stage of the trial. The median age was 53 yrs (range, 17-72). No objective responses have been noted. Six patients had stable disease after a median of 3.5 cycles of chemotherapy while 12 others had progressive disease. A total of 48 cycles were administered, 42 of which were administered at dose level 0 (4.5 mg/m2/d x 3). The median AGC nadir was 1.2/microl(0.1 -4.7) on day 10 and the median platelet nadir was 150,000/microl (31,000-338,000). Twenty cycles were complicated with grade 3-4 neutropenia and two cycles were complicated with FUO. There were no CNS toxicities. One patient had a grade 1 thrombophlebitis in 2 cycles and one other patient had a grade 4 thrombophlebitis in one cycle. In conclusion, CI-980 was well tolerated at this dose and schedule but inactive in soft-tissue sarcomas. PMID- 9740550 TI - Evaluation of pyrazoloacridine in patients with advanced pancreatic carcinoma. AB - PURPOSE: Pyrazoloacridine (PZA) is an acridine derivative selected for clinical development because of broad pre-clinical antitumor activity and solid tumor selectivity. Phase I evaluations with PZA have demonstrated predictable toxicity and suggested clinical efficacy. A phase II trial in patients with previously untreated advanced pancreatic cancer was conducted. METHODS: PZA was administered at a dose of 750 mg/m2 intravenously over 3 hours every 21 days. Seventeen patients were treated receiving a total of 46 courses of PZA. RESULTS: Of the 15 patients evaluable for response, no responses were observed (0% response rate, 95% confidence interval 0-22%). Major toxicities directly attributable to PZA included moderate neutropenia and mild neurotoxicity. CONCLUSION: PZA at this dose and schedule of administration was inactive in patients with pancreatic carcinoma. PMID- 9740551 TI - Systemic gene therapy with p53 inhibits breast cancer: recent advances and therapeutic implications. AB - Development of gene therapy technologies is approaching clinical realization for the treatment of neoplastic diseases. The use of tumor suppressor genes has been one useful strategy in gene therapy. Modifications and development of vectors as well as increased knowledge of the anti-tumor mechanisms of the p53 will play a significant role in the further advancement of this therapy. Currently, several laboratories have demonstrated that intratumoral injection of a virus carrying the p53 gene decreases tumor size in pre-clinical and clinical studies. Our lab has focused on a tumor-bearing mouse model in which intravenous delivery of liposome: p53 complexes decreases tumor growth. Although a high transfection efficiency of the tumor was thought to be necessary for gene therapy to exhibit anti-tumor activity with tumor suppressor genes, marked inhibition of the tumor occurs even with a low transfection efficiency. p53 may exhibit its bystander anti-tumor effect, at least in part, through an antiangiogenic effect. We believe that understanding the mechanism by which the p53 tumor suppressor gene inhibits tumor growth will lead to improvement in cancer therapy. PMID- 9740552 TI - The regulation of carbohydrate and fat metabolism during and after exercise. AB - The rate of carbohydrate utilization during prolonged, strenuous exercise is closely geared to the energy needs of the working muscles. In contrast, fat utilization during exercise is not tightly regulated, as there are no mechanisms for closely matching availability and metabolism of fatty acids to the rate of energy expenditure. As a result, the rate of fat oxidation during exercise is determined by the availability of fatty acids and the rate of carbohydrate utilization. Blood glucose and muscle glycogen are essential for prolonged strenuous exercise, and exhaustion can result either from development of hypoglycemia or depletion of muscle glycogen. Both absolute and relative (i.e. % of maximal O2 uptake) exercise intensities play important roles in the regulation of substrate metabolism. The absolute work rate determines the total quantity of fuel required, while relative exercise intensity plays a major role in determining the proportions of carbohydrate and fat oxidized by the working muscles. As relative exercise intensity is increased, there is a decrease in the proportion of the energy requirement derived from fat oxidation and an increase in that provided by carbohydrate oxidation. During moderately strenuous exercise of an intensity that can be maintained for 90 minutes or longer ( approximately 55-75% of VO2max), there is a progressive decline in the proportion of energy derived from muscle glycogen and a progressive increase in plasma fatty acid oxidation. The adaptations induced by endurance exercise training result in a marked sparing of carbohydrate during exercise, with an increased proportion of the energy being provided by fat oxidation. The mechanisms by which training decreases utilization of blood glucose are not well understood. However, the slower rate of glycogenolysis can be explained on the basis of lower concentrations of inorganic phosphate (Pi) in trained, as compared to untrained, muscles during exercise of the same intensity. The lower Pi level is a consequence of the increase in muscle mitochondria induced by endurance exercise training. A large increase in muscle glycogen concentration, far above the level found in the well-fed sedentary state, occurs in response to carbohydrate feeding following glycogen depleting exercise. It was recently found that this muscle "glycogen supercompensation" is markedly enhanced by endurance exercise training that induces an increase in the GLUT4 isoform of the glucose transporter in skeletal muscle. PMID- 9740553 TI - Molecular events in melanoma development and progression. AB - Based on clinical and histopathological features, five steps of melanoma progression have been proposed: common acquired and congenital nevi with structurally normal melanocytes, dysplastic nevus with structural and architectural atypia, early radial growth phase (RGP) primary melanoma, advanced vertical growth phase primary melanoma (VGP) with competence for metastasis, and metastatic melanoma. Despite a wealth of research resources (tissues, cell lines, and antibodies), the genetic alterations responsible for the development and stepwise progression of melanoma are still unclear. Cytogenetic analyses have failed to identify consistent gene deletions, mutations, translocations, or amplifications in sporadic cases. However, in vitro characterization of melanoma cells has revealed fundamental differences from normal melanocytes. Earlier work using monoclonal antibodies has defined a variety of melanoma-associated antigens that mediate cell-cell or cell-substratum adhesion, growth regulation, proteolysis, and modulation of immune responses. Functional studies of these individual candidate molecules will lead to a better understanding of the pathogenesis of melanoma and of potential targets for rational therapy. PMID- 9740554 TI - Apoptosis in the developing cerebellum of the thyroid hormone deficient rat. AB - The mechanism underlying transient reduction of cell number in the developing cerebellum have been studied for several decades. In this study we analyzed cell death by apoptosis in the developing cerebellum of euthyroid and hypothyroid rats. Results showed that in both groups the apoptotic activity is limited to the internal granular layer from postnatal (p) day 2 to day 12 in euthyroid animals, with the peak at 8 days. No apoptotic cells were detected in the cerebellum of 22 days old euthyroid rats. The level of apoptosis in the cerebellum of hypothyroid rats also reached a peak at 8 days but was four times higher than in control animals. Apoptosis in hypothyroid animals was also observed at p22 and corresponds to the value found in the time of the apoptotic peak in euthyroid cerebellum. At the age of 42 days, no apoptotic cells were found in the cerebellum of either group. Furthermore, it appears that the hormone also plays a role in the disappearance of the external germinal layer, since its presence is still apparent in 42 day old hypothyroid cerebellum. Hence, our results suggest that the deficiency of thyroid hormone (TH) not only increases, but also extends apoptosis during rat cerebellum development and affects the disappearance of the external germinal layer. PMID- 9740555 TI - Viagra: on release. Evidence on the effectiveness of sildenafil is good. PMID- 9740556 TI - Viagra and rationing. Let the sunlight in, let the people speak. PMID- 9740557 TI - Thalassaemia in Britain: a tale of two communities. Births are rising among British Asians but falling in Cypriots. PMID- 9740559 TI - An amnesty for unpublished trials. One year on, many trials are unregistered and the amnesty remains open. PMID- 9740558 TI - Implantable defibrillators for life threatening ventricular arrhythmias. Are more effective than antiarrhythmic drugs in selected high risk patients. PMID- 9740560 TI - Action on antimicrobial resistance. Not easy, but Europe can do it. PMID- 9740561 TI - Preferences for chemotherapy in patients with advanced non-small cell lung cancer: descriptive study based on scripted interviews. AB - OBJECTIVE: To determine how patients with lung cancer value the trade off between the survival benefit of chemotherapy and its toxicities. DESIGN: Scripted interviews that included three hypothetical scenarios. Each scenario described the same patient with metastatic non-small cell lung cancer with an expected survival of 4 months without treatment. Subjects were asked to indicate the minimum survival benefit required to accept the side effects of chemotherapy in the first two scenarios (mild toxicity and severe toxicity). In the third scenario, subjects were asked to choose between chemotherapy and supportive care when the benefit of chemotherapy was either to prolong life by 3 months or to palliate symptoms. SUBJECTS: 81 patients previously treated with cis-platinum based chemotherapy for advanced non-small cell lung cancer. MAIN OUTCOME MEASURE: Survival threshold for accepting chemotherapy. RESULTS: The minimum survival threshold for accepting the toxicity of chemotherapy varied widely in patients. Several patients would accept chemotherapy for a survival benefit of 1 week, while others would not choose chemotherapy even for a survival benefit of 24 months. The median survival threshold for accepting chemotherapy was 4.5 months for mild toxicity and 9 months for severe toxicity. When given the choice between supportive care and chemotherapy only 18 (22%) patients chose chemotherapy for a survival benefit of 3 months; 55 (68%) patients chose chemotherapy if it substantially reduced symptoms without prolonging life. CONCLUSIONS: Patients' willingness to accept chemotherapy for the treatment of metastatic lung cancer varies widely. Many would not choose chemotherapy for its likely survival benefit of 3 months but would if it improved quality of life. The conflict between these patients' preferences and the care they previously received has several explanations, one being that some patients had not received the treatment they would have chosen had they been fully informed. PMID- 9740562 TI - Working hours as a risk factor for acute myocardial infarction in Japan: case control study. AB - OBJECTIVE: To clarify the extent to which working hours affect the risk of acute myocardial infarction, independent of established risk factors and occupational conditions. DESIGN: Case-control study. SETTING: University and general hospitals and routine medical examinations at workplaces in Japan. SUBJECTS: Cases were 195 men aged 30-69 years admitted to hospital with acute myocardial infarction during 1990-3. Controls were 331 men matched at group level for age and occupation who were judged to be free of coronary heart diseases at routine medical examinations in the workplace. MAIN OUTCOME MEASURES: Odds ratios for myocardial infarction in relation to previous mean daily working hours in a month and changes in mean working hours during previous year. RESULTS: Compared with men with mean working hours of >7-9 hours, the odds ratio of acute myocardial infarction (adjusted for age and occupation) for men with working hours of >11 hours was 2.44 (95% confidence interval 1.26 to 4.73) and for men with working hours of <=7 hours was 3.07 (1.77 to 5.32). Compared with men who experienced an increase of <=1 hour in mean working hours, the adjusted odds ratio of myocardial infarction for men who experienced an increase of >3 hours was 2.53 (1.34 to 4. 77). No appreciable change was observed when odds ratios were adjusted for established and psychosocial risk factors for myocardial infarction. CONCLUSION: There was a U shaped relation between the mean working hours and the risk of acute myocardial infarction. There also seemed to be a trend for the risk of infarction to increase with greater increases in mean working hours. PMID- 9740564 TI - Never again PMID- 9740563 TI - As seen on TV: observational study of cardiopulmonary resuscitation in British television medical dramas. AB - OBJECTIVE: To determine the frequency and accuracy with which cardiopulmonary resuscitation is portrayed in British television medical dramas. DESIGN: Observational study. SUBJECTS: 64 episodes of three major British television medical dramas: Casualty, Cardiac Arrest, and Medics. MAIN OUTCOME MEASURES: Frequency of cardiopulmonary resuscitation shown on television; age, sex, and diagnosis of the patients undergoing resuscitation; rate of survival through resuscitation. RESULTS: Overall 52 patients had a cardiorespiratory arrest on screen and 3 had a respiratory arrest alone, all the arrests occurring in 40 of the 64 episodes. Of the 52 patients having cardiorespiratory arrest, 32 (62%) underwent an attempt at cardiopulmonary resuscitation; 8 attempts were successful. All 3 of the patients having respiratory arrests alone received ventilatory support and survived. On 48% of occasions, victims of cardiac arrest seemed to be less than 35 years old. CONCLUSIONS: Cardiorespiratory resuscitation is often depicted in British television medical dramas. Patients portrayed receiving resuscitation are likely to be in a younger age group than in real life. Though the reasons for resuscitation are more varied and more often associated with trauma than in reality, the overall success rate is nevertheless realistic. Widespread overoptimism of patients for survival after resuscitation cannot necessarily be blamed on British television medical dramas. PMID- 9740565 TI - Dietary intake of schizophrenic patients in Nithsdale, Scotland: case-control study. PMID- 9740566 TI - Effect of fundholding on removing patients from general practitioners' lists: retrospective study. PMID- 9740567 TI - Purpuric rash with donepezil treatment. PMID- 9740568 TI - Withdrawal reaction associated with venlafaxine. PMID- 9740569 TI - A multidisciplinary approach for improving services in primary care: randomised controlled trial of screening for haemoglobin disorders. AB - OBJECTIVE: To investigate the feasibility of improving screening for carriers of haemoglobin disorders in general practice by using a nurse facilitator to work with primary care teams and the relevant haematology laboratories; to identify problems in communication between all those involved in delivering the service, and to implement solutions. DESIGN: Two year, practice based randomised controlled trial. SETTING: North London area where 29% of residents and 43% of births are in ethnic groups at risk for haemoglobin disorders. SUBJECTS: 26 of the 93 practices using the services of the area's haematology laboratory agreed to take part and were randomly divided into control and intervention practices. MAIN OUTCOME MEASURE: Change in number of requests for screening tests for haemoglobin disorders made by control and intervention practices in baseline and intervention years. RESULTS: The number of screening tests requested varied from 0-150 in the 93 practices in the baseline year. Study practices tended to have made a moderate number of requests (10-50) during this period. During the intervention year intervention practices made 292 more requests (99% increase) and control practices made 74 fewer requests (23% decrease; P=0.001 for difference in median change). Four practices, three of which were singlehanded, accounted for 75% of the increase. The number of requests from intervention practices, adjusted for baseline requests, was 3.2 times higher than control practices (P<0.0001). CONCLUSION: General practitioners and practice nurses are willing to undertake a new genetic screening service (or expand an existing one) if they are persuaded that it benefits the health of a significant proportion of their practice population. They need appropriate tools (for example, information materials for carriers and groups at risk), and the laboratory must be sensitive to their needs. Preconceptional carrier screening and counselling need to be coupled with antenatal screening. PMID- 9740570 TI - Recent advances. General medicine. PMID- 9740572 TI - The white album PMID- 9740571 TI - Late onset interstitial nephritis associated with mesalazine treatment. PMID- 9740573 TI - ABC of oxygen. Acute oxygen therapy. PMID- 9740575 TI - Competency, consent, and the duty of care: ethical dilemma. PMID- 9740577 TI - Breaking down language barriers . Some ethnic groups may have problems in getting as far as a consultation PMID- 9740576 TI - The aftermath of the bristol case . Case arose through a failure of action, not of detection PMID- 9740578 TI - Pressure on doctors to prescribe needs measuring directly PMID- 9740579 TI - Safety of patients participating in drug trials . Use Of placebo in trials of drugs for mental illness should be debated PMID- 9740574 TI - North of England evidence based guidelines development project: guideline for the primary care management of dementia. PMID- 9740580 TI - If devolved facilities are used, processes may be streamlined PMID- 9740581 TI - Use of the READER method of critical appraisal in general practice . Study did not properly answer the questions it posed PMID- 9740582 TI - Living wills might make patients at risk of death by starvation and dehydration PMID- 9740583 TI - Deaths outside hospital from acute coronary events . Early access to defibrillation is a key to survival PMID- 9740584 TI - Drug treatments for asthma may cause erosive tooth damage PMID- 9740585 TI - Susan elizabeth jean ("Sue") munby PMID- 9740586 TI - Evaluation and management guidelines PMID- 9740587 TI - The discovery of cortisone: a personal memory PMID- 9740588 TI - The rise and fall of viagra. PMID- 9740589 TI - Before I say goodbye PMID- 9740590 TI - The human rights, ethical and moral dimensions of health care: 120 practical case studies PMID- 9740592 TI - Lung cancer patients vary in their preferences for chemotherapy PMID- 9740591 TI - A corker of a journal? PMID- 9740594 TI - Patients with schizophrenia living in the community eat badly PMID- 9740593 TI - Working hours are associated with acute myocardial infarction in japan PMID- 9740595 TI - British television medical dramas realistically depict cardiopulmonary resuscitation PMID- 9740597 TI - What to do with a neglectful patient who refuses surgery PMID- 9740596 TI - Small practices are ready to take on screening for haemoglobin disorders PMID- 9740598 TI - AHA journals lead with definitive new online site. PMID- 9740599 TI - Transcription-modulating drugs: a new frontier in the treatment of essential hypertension. AB - While the promises of gene therapy may be years away from realization, the therapeutic use of drugs that act by modifying gene transcription is a well established practice in clinical medicine. Although transcription-modulating drugs are frequently used in many different specialties, the deliberate development and use of these agents in cardiovascular medicine has been comparatively limited. However, research advances in the area of gene transcription and in the molecular genetic regulation of blood pressure, insulin resistance, lipid metabolism, and cell growth are providing new opportunities for controlling the expression of genes that are relevant to the pathogenesis of cardiovascular disease and essential hypertension. These research advances are beginning to converge in the development of transcription-modulating drugs with the potential to attack genetically determined risk factors that often cluster in patients with essential hypertension. Ligand-activated transcription factors that serve as receptors for small lipophilic compounds such as the thiazolidinediones and retinoids represent examples of potential therapeutic targets with direct effects on the expression of genes relevant to the pathogenesis of essential hypertension and its complications. Mounting evidence suggesting that the superior cardiorenal protective properties of converting enzyme inhibitors are related in part to their ability to indirectly modify the expression of genes in the heart and vasculature provides provisional support for the clinical value of this therapeutic approach. Given the success of transcription-modulating drugs in the treatment of type II diabetes and many other clinical disorders, it is anticipated that these agents will be developed as tools for the prevention and treatment of hypertension and cardiovascular disease in the not too distant future. PMID- 9740600 TI - Pathways for angiotensin II generation in intact human tissue: evidence from comparative pharmacological interruption of the renin system. AB - Multiple lines of evidence have suggested that alternative pathways to the angiotensin-converting enzyme (ACE) exists for angiotensin II (Ang II) generation in the heart, large arteries, and the kidney. In vitro studies in intact tissues, homogenates, or membrane isolates from the heart and large arteries have repeatedly demonstrated such pathways, but the issue remains unresolved because the approaches used have not made it possible to extrapolate from the in vitro to the in vivo situation. For our in vivo model, we studied young and healthy human volunteers, for the most part white and male; when these subjects achieved balance on a low salt diet to activate the renin system, the response of renal perfusion to pharmacological interruption of the renin system was studied. With this approach, we studied the renal vasodilator response to 3 ACE inhibitors, 2 renin inhibitors, and 2 Ang II antagonists at the top of their respective dose response relationships. When these studies were initiated, our premise was that a kinin-dependent mechanism contributed to the renal hemodynamic response to ACE inhibition; therefore, the renal vasodilator response to ACE inhibition would exceed the alternatives. To our surprise, both renin inhibitors and both Ang II antagonists that were studied induced a renal vasodilator response of 140 to 150 mL/min/1.73 m2, approximately 50% larger than the maximal renal hemodynamic response to ACE inhibition, which was 90 to 100 mL/min/1.73 m2. In light of the data from in vitro systems, our findings indicate that in the intact human kidney, virtually all Ang II generation is renin-dependent but at least 40% of Ang I is converted to Ang II by pathways other than ACE, presumably a chymase, although other enzyme pathways exist. Preliminary data indicate that the non-ACE pathway may be substantially larger in disease states such as diabetes mellitus. One implication of the studies is that at the tissue level, Ang II antagonists have much greater potential for blocking the renin-angiotensin system than does ACE inhibition-with implications for therapeutics. PMID- 9740601 TI - Angiotensinogen genotype, sodium reduction, weight loss, and prevention of hypertension: trials of hypertension prevention, phase II. AB - The angiotensinogen gene has been linked to essential hypertension and increased blood pressure. A functional variant believed to be responsible for hypertension susceptibility occurs at position -6 in the promoter region of the gene in which an A for G base pair substitution is associated with higher angiotensinogen levels. To test whether an allele within the angiotensinogen gene is related to subsequent incidence of hypertension and blood pressure response to sustained sodium reduction, 1509 white male and female subjects participating in phase II of the Trials of Hypertension Prevention were genotyped at the angiotensinogen locus. Participants had diastolic blood pressures between 83 and 89 mm Hg and were randomized in a 2x2 factorial design to sodium reduction, weight loss, combined intervention, or usual care groups. Persons in the usual care group with the AA genotype at nucleotide position -6 had a higher 3-year incidence rate of hypertension (44.6%) compared with those with the GG genotype (31.5%), with a relative risk of 1.4 (95% confidence interval [0.87, 2.34], test for trend across all 3 genotypes, P=0.10). In contrast, the incidence of hypertension was significantly lower after sodium reduction for persons with the AA genotype (relative risk=0.57 [0.34, 0.98] versus usual care) but not for persons with the GG genotype (relative risk=1.2 [0.79, 1.81], test for trend P=0.02). Decreases of diastolic blood pressure at 36 months in the sodium reduction group versus usual care showed a significant trend across all 3 genotypes (P=0.01), with greater net blood pressure reduction in those with the AA genotype (-2.2 mm Hg) than those with the GG genotype (+1.1 mm Hg). A similar trend across the 3 genotypes for net systolic blood pressure reduction (-2.7 for AA versus -0.2 mm Hg for GG) was not significant (P=0.17). Trends across genotypes for the effects of weight loss on hypertension incidence and decreases in blood pressure were similar to those for sodium reduction. We conclude that the angiotensinogen genotype may affect blood pressure response to sodium or weight reduction and the development of hypertension. PMID- 9740602 TI - Angiotensinogen genotype and blood pressure responses to reduced dietary NaCl and to weight loss. PMID- 9740603 TI - A population-based study on blood pressure and brain atrophy in 85-year-olds. AB - In the general population, mean systolic and diastolic blood pressure increases up to age 75 years but decreases thereafter. The brain has a role in blood pressure regulation; it is not clear whether the cerebral changes that occur with aging contribute to the decline in blood pressure in the very elderly. We examined a population-based sample of 484 85-year-old persons (344 nondemented and 140 demented, 61 with Alzheimer's disease, 65 with vascular dementia, and 14 with other types of dementia) with a neuropsychiatric examination and blood pressure measurements. Dementia was diagnosed according to the criteria proposed in the Diagnostic and Statistical Manual of Mental Disorders, edition 3, revised. Brain atrophy was measured by CT of the brain. In the nondemented group, frontal (r=-0.18, P=0.037) and parietal (r=-0.23, P=0.008) cortical atrophy and bifrontal ratio (r=-0.20, P=0.013) were associated with lower systolic blood pressure, and frontal (r=-0.23, P=0.010) and parietal (r=-0.24, P=0.008) cortical atrophy and bifrontal ratio (r=-0.23, P=0.006) with lower diastolic blood pressure. Systolic blood pressure was lower in subjects with Alzheimer's disease and vascular dementia, and diastolic blood pressure was lower in those with vascular dementia compared with the nondemented. Systolic (r=-0.27, P<0.0001) and diastolic (r= 0.10, P=0.020) blood pressure was negatively correlated to dementia severity. In the demented subjects, frontal cortical atrophy was correlated to lower diastolic blood pressure (r=-0.21, P=0.043). Our findings suggest that age-related changes in brain structure may contribute to the decrease in blood pressure in the very elderly and that low blood pressure in dementia disorders is mainly a secondary phenomenon. PMID- 9740604 TI - Calcium channel blockade and cardiovascular prognosis in the European trial on isolated systolic hypertension. AB - In the double-blind Systolic Hypertension in Europe (Syst-Eur) Trial, active treatment was initiated with nitrendipine (10 to 40 mg/d) with the possible addition of enalapril (5 to 20 mg/d) and/or hydrochlorothiazide (12.5 to 25 mg/d) titrated or combined to reduce sitting systolic blood pressure by at least 20 mm Hg to <150 mm Hg. In the control group, matching placebos were used similarly. In view of persistent concerns about the use of calcium channel blockers as first line antihypertensive drugs, this report explored to what extent nitrendipine, administered alone, prevented cardiovascular complications. Age at randomization averaged 70.2 years and systolic/diastolic blood pressure 173.8/85.5 mm Hg. Of 2398 actively treated patients, 1327 took only nitrendipine (average dose, 23.4 mg/d), and 1042 progressed to other treatments including nitrendipine (n=757; 35.7 mg/d), enalapril (n=783; 13.4 mg/d), and/or hydrochlorothiazide (n=294; 21.0 mg/d). Compared with the whole placebo group (n=2297), patients receiving monotherapy with nitrendipine had 25% (P=0.05) fewer cardiovascular end points, and those progressing to other active treatments showed decreases (P/=0.5 or <0.5, the regression of LVMI was similar. In conclusion, peak and trough blood pressure changes are reproducible and predict the regression of LVMI induced by treatment as well as average 24-hour blood pressure. T/Ps are less reproducible, and their value does not predict regression of organ damage by antihypertensive treatment. PMID- 9740605 TI - Impact of shift work and race/ethnicity on the diurnal rhythm of blood pressure and catecholamines. AB - To evaluate the effects of shift work and race/ethnicity on the diurnal rhythm of blood pressure and urinary catecholamine excretion of healthy female nurses, 37 African American women and 62 women of other races underwent ambulatory blood pressure monitor and urine collection for 24 hours that included a full work shift: day shift (n=61), evening shift (n=11), and night shift (n=27). Awake and sleep times were evaluated from subjects' diaries. Of African Americans, 79% who were working evenings or nights and 32% working day shifts were nondippers (<10% drop in systolic pressure during sleep), whereas only 29% of others working evening+night and 8% working day shifts were nondippers. Regression analyses indicated that evening+night shift workers had a 5.4 mm Hg (P<0.001) smaller drop than day shift workers, and African Americans had a 4.0 mm Hg (P<0.01) smaller drop than others. The odds of an evening+night shift worker being a nondipper were 6.1 times that of a day shift worker (P<0.001), and the odds of an African American were 7.1 times that of others (P<0.001). Total sleep time was significantly greater in the non-African American day shift workers than in the other 3 groups. After controlling for work shift and race/ethnicity, we determined that longer sleep times predicted less dipping (absolute and relative) in blood pressure. Urinary norepinephrine and epinephrine were higher during work than nonwork in both racial groups of day shift workers, but in evening+night shift workers the difference was small and in the opposite direction. These results indicate that being African American and working evening or night shifts are independent predictors of nondipper status. Higher sleep blood pressure may contribute to the known adverse effects of shift work. PMID- 9740607 TI - Statistical base value of 24-hour blood pressure distribution in patients with essential hypertension. AB - The purpose of this study was to calculate statistically the minimum (base) blood pressure (BP) of nighttime (sleep-time) BP values obtained by ambulatory BP monitoring (ABPM) and to investigate its clinical significance. Twenty-four-hour recording of ECG with ABPM was performed directly (n=89) or indirectly (n=117) in 206 patients with essential hypertension. A telemeter was used for the direct method and a multi-biomedical recorder (TM2425) was used for indirect measurement. First, minimum heart rate (HR0=60/RR0) was determined from sleep time ECG. The mean product of sleep-time diastolic BP (DBP) and pulse interval (RR) was divided by RR0 to obtain DBP0 [DBP0=(DBPxRR)s/RR0]. The correlation between systolic BP (SBP) and DBP was used to determine SBP0 corresponding to DBP0. Statistical base mean BP (MBP0) was calculated from these values, and its reproducibility and relation to hypertension severity were investigated. MBP0 values were similar to true base values of sleep-time MBP obtained by the direct method (mean+/-SD difference, 2.0+/-4.2 mm Hg). Direct MBP0 criteria predicted hypertension severity (mild, moderate, or severe target organ damage) more accurately (predictive accuracy, 89%) than daytime MBP criteria (53%, P<0.01). Almost the same results were obtained using indirect MBP0 criteria. Day-to-day indirect MBP0 variation (mean absolute difference) was smaller (2.4+/-1.8 mm Hg) than day-to-day daytime and nighttime MBP variation (6.3+/-5.3 and 5.4+/-3.4 mm Hg, respectively; n=61, P<0.01), and the correlation coefficient between day-to day variations of daytime MBP and physical activity (measured by an acceleration sensor) was 0.38 (P<0.05). In conclusion, statistical base BP was almost equal to true base (minimum) BP of sleep-time BP distribution. It was closely related to the severity of hypertensive organ damage, was highly reproducible, and is considered likely to serve stochastically and physiologically as a representative BP value in an individual subject. PMID- 9740608 TI - Large artery remodeling during aging: biaxial passive and active stiffness. AB - To examine arterial mechanical changes during aging, pressure-radius and axial force-radius curves were measured in vivo in carotid arteries from 6- and 23 month-old Brown Norway X Fischer 344 rats. Incremental passive circumferential stiffness (measured at 50, 100, and 200 mm Hg) was higher (P<0.01) in the 23- compared with the 6-month-old rats (14.02+/-1.23 versus 6.58+/-1.51; 2.68+/-0.56 versus 0.99+/-0.34; 1.10+/-0.24 versus 0.69+/-0.15 dyne/mm2x10(3), respectively). Incremental passive axial stiffness was increased (P<0.01) in the 23- compared with the 6-month-old rats (7.95+/-0.70 versus 4.24+/-0.81; 1.91+/-0.10 versus 0.61+/-0.16; 0.58+/-0.09 versus 0.36+/-0.06 dyne/mm2x10(3), respectively). Active incremental circumferential arterial stiffness at 100 and 200 mm Hg was increased (P<0.01) in the older rats. In 6-month-old rats, activation of vascular smooth muscle enhanced (P<0.01) the incremental circumferential and axial stiffness measured at 200 mm Hg. In 23-month-old rats, only active incremental stiffness was increased (P<0.01) at 200 mm Hg. Aging increased (P<0.05) media thickness, collagen content, and the collagen/elastin ratio by 12%, 21%, and 38%, respectively. Elastin density and the number of smooth muscle cell nuclei were decreased by 20% and 31%, respectively, with aging. Thus, structural alterations that occur with aging are associated with changes in both active and passive stiffness. Vascular smooth muscle tone modulates arterial wall anisotropy differently during aging. PMID- 9740610 TI - Pressure mediates angiotensin II-induced arterial hypertrophy and PDGF-A expression. AB - Angiotensin II (Ang II) may induce arterial hypertrophy either directly or through an increase in arterial pressure. To separate these 2 mechanisms, rats were implanted with osmopumps delivering either Ang II (100 ng x kg-1 x min-1) or saline. 5-Bromo-2'-deoxyuridine (BrdU) was delivered to both groups by osmopump (2.5 microg x kg-1 x min-1). Half of the rats in each group were given minoxidil (9 mg x kg-1 . d-1) in their drinking water. After 14 days, systolic blood pressure was 117+/-2, 124+/-3, and 115+/-2 mm Hg in the control, Ang II minoxidil, and minoxidil groups, respectively, and 181+/-6 mm Hg in the Ang II group (P<0.05). After perfusion-fixation, the thoracic aorta, carotid artery, small mesenteric artery, external spermatic artery, and kidneys were harvested, paraffin-embedded, and used for morphological measurements, immunohistochemistry for BrdU, and in situ hybridization with a 35S-labeled riboprobe for platelet derived growth factor-A chain (PDGF-A) mRNA. The walls of the aorta and carotid arteries hypertrophied in the Ang II group only. There were no significant morphological differences in the small arteries. BrdU was negative in all arteries but positive in the renal tubules. Expression of PDGF-A was elevated 8 fold in the thoracic aorta of the Ang II group (P<0.05). These results show that (1) arterial hypertrophy from Ang II infusion occurs in response to elevated arterial pressure, (2) hypertrophy was not associated with hyperplasia or polyploidy of vascular smooth muscle cells, and (3) PDGF-A expression correlated with elevated pressure and arterial wall hypertrophy. PMID- 9740609 TI - Negative regulation of local hepatocyte growth factor expression by angiotensin II and transforming growth factor-beta in blood vessels: potential role of HGF in cardiovascular disease. AB - Because hepatocyte growth factor (HGF) is a member of the endothelium-specific growth factors, we hypothesized that HGF may play a role in cardiovascular disease. Therefore we first examined the role of local HGF production in endothelial cell (EC) growth. Addition of anti-HGF antibody to EC resulted in a significant decrease in EC number. Moreover, coculture of vascular smooth muscle cells (VSMC) with EC resulted in an increase in EC number that was completely inhibited by anti-HGF antibody, suggesting that HGF secreted from EC and VSMC regulates EC growth in an autocrine-paracrine manner. Interestingly, transforming growth factor (TGF)-ss significantly decreased HGF secretion from EC, whereas interleukin 6 stimulated immunoreactive HGF secretion. In human VSMC, TGF-ss and angiotensin II suppressed local HGF production in a dose-dependent manner. Interestingly, anti-TGF-beta antibody resulted in significant but not complete inhibition of the decrease in local HGF production. To further study the regulation of local HGF production, we used a coculture system. Coculture of VSMC with EC resulted in a significant decrease in local HGF secretion. The decrease in local HGF production by coculture was significantly attenuated by anti-TGF beta antibody, suggesting that inhibition of local HGF production in the coculture system was due to TGF-beta activation. Moreover, a further decrease in local HGF production in the coculture system by angiotensin II was also observed. Finally, we studied the role of angiotensin II in the regulation of the local HGF system in vivo by using a balloon injury rat model. Of importance, local HGF production was significantly decreased in balloon-injured arteries compared with intact vessels, accompanied by a reduction of HGF mRNA. An angiotensin-converting enzyme inhibitor (cilazapril) or an angiotensin II type 1 receptor antagonist (E 4177) significantly stimulated local vascular HGF production associated with the inhibition of neointimal formation after balloon injury compared with vehicle. In contrast, hydralazine did not alter local HGF production or neointimal formation despite decreasing blood pressure to a similar level as that in rats treated with cilazapril or E-4177. Overall, local HGF secretion from vascular cells was negatively regulated by TGF-beta and angiotensin II. The present study also demonstrated that blockade of angiotensin II significantly inhibited neointimal formation, accompanied by a significant increase in local vascular HGF production in vivo in the balloon injury model. Given the strong mitogenic activity of HGF on endothelial cells, increased local HGF production by blockade of angiotensin II may enhance reendothelialization after balloon injury. Downregulation of the local vascular HGF system by TGF-beta and vascular angiotensin may play an important role in the pathogenesis of cardiovascular diseases. PMID- 9740611 TI - Angiotensin II type 1 receptor: relationship with caveolae and caveolin after initial agonist stimulation. AB - Caveolae are membrane domains that have been implicated in signal transduction, and caveolins are major structural components of these domains. We found that all reported caveolin isoforms (caveolin-1, -2, and -3) were expressed in vascular smooth muscle cells (VSMCs); however, only caveolin-1 mRNA was regulated by angiotensin II (Ang II). Ang II (100 nmol/L) increased caveolin-1 mRNA, with a peak at 2 hours (193+/-6% of control, P<0.01, n=4). In contrast, Ang II significantly decreased caveolin-1 protein, with a nadir at 4 hours (64+/-5% of control, P<0.01, n=6). [35S]Methionine labeling showed that Ang II increased caveolin biosynthesis (226+/-33% of control labeling at 4 hours), suggesting that the transient decrease in caveolin protein levels is due to increased degradation. When cells were fractionated with sucrose, on agonist stimulation, AT1 receptors appeared in fraction 5 where caveolin was fractionated. This migration was blocked by low temperature and treatment with phenylarsine oxide, interventions that interfere with agonist-induced Ang II type 1 (AT1) receptor sequestration and tonic phase signaling. In addition, caveolin-1 coimmunoprecipitates with AT1 receptor only on agonist stimulation. These data support the concept that the caveola is a specialized signaling domain in VSMCs that can be dynamically accessed by the AT1 receptor. Because of the signaling and coupling proteins that are localized in caveolae and because of evidence that these proteins may interact directly with caveolin, caveola-AT1 receptor interaction likely represents an important focus for dynamic control of receptor signaling in VSMCs. PMID- 9740612 TI - Interaction of mRNAs for angiotensin II type 1 and type 2 receptors to vascular remodeling in spontaneously hypertensive rats. AB - We administered angiotensin II (Ang II) receptor type 1 (AT1) blockade (losartan, 40 mg x kg-1 x d-1), type II receptor (AT2) blockade (PD123319, 100 mg x kg-1 x d 1), or angiotensin-converting enzyme (ACE) inhibitor (enalapril, 30 mg x kg-1 x d 1) to spontaneously hypertensive rats (SHR) from 10 to 20 weeks of age. Control SHR and Wister-Kyoto rats (WKY) received a placebo for the same period. At the end of treatment, losartan and enalapril were both found to have significantly reduced the arterial systolic blood pressure and the collagen concentration to the level of WKY, whereas PD123319 had no effect. Enalapril and PD123319 significantly reduced the media cross-sectional area of the aorta in comparison to that of untreated SHR, which was still larger than that of the WKY; however, losartan did not change it. Using reverse transcription-polymerase chain reaction, we next examined the mRNA expressions for ACE, AT1 receptor, and AT2 receptor in experimental animals. We observed significantly enhanced mRNA expression for AT1 and AT2 receptors and ACE in untreated SHR compared with WKY. The AT1 mRNA level was also significantly decreased in the SHR treated with either losartan or enalapril, whereas the AT2 mRNA level was significantly decreased in the SHR treated with either PD123319 or enalapril in comparison to untreated SHR. The level of ACE mRNA was significantly decreased only in the SHR treated with enalapril. These results indicate that AT1 receptor, but not AT2 receptor, plays a crucial role in the remodeling of matrix tissue, while AT2 receptor may play a role in the development of hypertrophy of smooth muscle in aorta in SHR, and that the reduction of hypertrophy of smooth muscle does not fully account for the suppression of hypertension. PMID- 9740613 TI - MAP kinase-independent signaling in angiotensin II regulation of neuromodulation in SHR neurons. AB - Angiotensin II (Ang II), via its interaction with the angiotensin type 1 (AT1) receptor subtype, causes enhanced stimulation of norepinephrine (NE) neuromodulation. This involves increased transcription of NE transporter, tyrosine hydroxylase, and dopamine ss-hydroxylase genes in Wistar-Kyoto rat (WKY) brain neurons. AT1 receptor-mediated regulation of certain signaling events (such as activation of the Ras-Raf-1-mitogen activated protein (MAP) kinase signaling pathway, nuclear translocation of transcription factors such as Fos and Jun, and the interactions of these factors with AP-1 binding sites) is involved in this NE neuromodulation (Lu et al. J Cell Biol. 1996;135:1609-1617). The aim of this study was to compare the signal transduction mechanism of Ang II regulation of NE neuromodulation in WKY and spontaneously hypertensive rat (SHR) brain neurons, in view of the fact that AT1 receptor expression and Ang II stimulation of NE neuromodulation are higher in SHR neurons compared with WKY neurons. Despite this hyperactivity, Ang II stimulation of Ras, Raf-1, and MAP kinase activities was comparable between the neurons from WKY and SHR. Similarly, central injections of Ang II caused a comparable stimulation of MAP kinase in the hypothalamic and brain stem areas of adult WKY and SHR. Inhibition of MAP kinase by either an MAP kinase kinase inhibitor (PD98059) or an MAP kinase antisense oligonucleotide completely attenuated the stimulatory effects of Ang II on [3H]-NE uptake, NE transporter mRNA, and tyrosine hydroxylase mRNA levels in WKY neurons. These treatments resulted in only 43% to 50% inhibition of [3H]-NE uptake and NE transporter and tyrosine hydroxylase mRNAs in SHR neurons. Thus, Ang II stimulation of NE neuromodulation was completely blocked by MAP kinase inhibition in WKY neurons and only partially blocked in the SHR neurons. These observations suggest the presence of an additional signal transduction pathway involved in NE neuromodulation in SHR neurons that is independent of the MAP kinase pathway. PMID- 9740614 TI - Insulin-mediated growth in aortic smooth muscle and the vascular renin angiotensin system. AB - Insulin has been shown to directly affect blood vessel tone and to promote vascular hypertrophy, but the mechanism of these actions remains uncertain. Because angiotensin I (Ang I)-converting enzyme inhibitors have been shown to improve insulin action and to impede the progression of vascular hypertrophy in hypertensive animal models, it is possible that the vascular properties of insulin may be mediated through the tissue renin-angiotensin system (RAS). To evaluate this relationship, we first investigated the effect of insulin on components of the RAS using cultured rat vascular smooth muscle cells (VSMCs). Insulin treatment (1000 microU/mL) markedly increased angiotensinogen mRNA expression and angiotensinogen production. We next investigated the role of the RAS in insulin-mediated cell proliferation, using [3H]thymidine uptake. Studies were done both with insulin alone and in the presence of captopril (1x10(-7) to 10(-5) mol/L) and losartan (1x10(-9) to 10(-7) mol/L). [3H]Thymidine uptake was increased significantly by 1000 microU/mL insulin, and this stimulation was reduced by 1x10(-6) mol/L captopril (-38.8%, P<0.05) and by 1x10(-8) mol/L losartan (-37. 5%, P<0.05). Further studies showed that the degree of insulin mediated [3H]thymidine uptake in VSMCs could be duplicated by 4x10(-10) mol/L Ang II. Losartan reduced the effects of both Ang II and insulin on [3H]thymidine uptake by about 40% to 45% of baseline (P<0.05). Captopril reduced insulin mediated [3H]thymidine uptake but did not affect Ang II-mediated [3H]thymidine uptake. In summary, insulin induced significant stimulation of angiotensinogen expression and production and stimulated growth similar to that seen with Ang II in cultured rat VSMCs. Inhibition of Ang II production or its binding to the Ang II type 1 (AT1) receptor inhibited insulin-mediated growth in a fashion similar to that seen with inhibition of Ang II-mediated growth. Thus, insulin can modulate the vascular RAS, and the effect of insulin on vascular growth may be via direct effects on angiotensinogen expression and translation operative through both the AT1 receptor and the conversion of Ang I to Ang II. PMID- 9740616 TI - Converting enzyme determines plasma clearance of angiotensin-(1-7). AB - We determined the mechanism accounting for the removal and metabolism of angiotensin-(1-7) [Ang-(1-7)] in 21 anesthetized spontaneously hypertensive (SHR), 18 age-matched normotensive Sprague-Dawley (SD), and 36 mRen-2 transgenic (TG+) rats. Animals of all 3 strains were provided with tap water or tap water containing losartan, lisinopril, or a combination of lisinopril and losartan for 2 weeks. On the day of the experiment, Ang-(1-7) was infused for a period of 15 minutes at a rate of 278 nmol . kg-1 . min-1. After this time, samples of arterial blood were collected rapidly at regular intervals for the assay of plasma Ang-(1-7) levels by radioimmunoassay. Infusion of Ang-(1-7) had a minimal effect on vehicle-treated SD rats but elicited a biphasic pressor/depressor response in vehicle-treated SHR and TG+ rats. In lisinopril-treated rats, Ang-(1 7) infusion increased blood pressure, whereas losartan treatment abolished the pressor component of the response without altering the secondary fall in arterial pressure. Combined treatment with lisinopril and losartan abolished the cardiovascular response to Ang-(1-7) in all 3 strains. In vehicle-treated SD, SHR and TG+ the half-life (t1/2) of Ang-(1-7) averaged 10+/-1, 10+/-1, and 9+/-1 seconds, respectively. Lisinopril alone or in combination with losartan produced a statistically significant rise in the half-life of Ang-(1-7) in all 3 strains of rats. Plasma clearance of Ang-(1-7) was significantly greater in the untreated SD rats compared with either the SHR or TG+ rat. Lisinopril treatment was associated with reduced clearance of Ang-(1-7) in all 3 strains. Concurrent experiments in pulmonary membranes from SD and SHR showed a statistically significant inhibition of 125I-Ang-(1-7) metabolism in the presence of lisinopril. These studies showed for the first time that the very short half-life of Ang-(1-7) in the circulation is primarily accounted for peptide metabolism by ACE. These findings suggest a novel role of ACE in the regulation of the production and metabolism of the two primary active hormones of the renin angiotensin system. PMID- 9740615 TI - Role of NADH/NADPH oxidase-derived H2O2 in angiotensin II-induced vascular hypertrophy. AB - Recent evidence suggests that oxidative mechanisms may be involved in vascular smooth muscle cell (VSMC) hypertrophy. We previously showed that angiotensin II (Ang II) increases superoxide production by activating an NADH/NADPH oxidase, which contributes to hypertrophy. In this study, we determined whether Ang II stimulation of this oxidase results in H2O2 production by studying the effects of Ang II on intracellular H2O2 generation, intracellular superoxide dismutase and catalase activity, and hypertrophy. Ang II (100 nmol/L) significantly increased intracellular H2O2 levels at 4 hours. Neither superoxide dismutase activity nor catalase activity was affected by Ang II; the SOD present in VSMCs is sufficient to metabolize Ang II-stimulated superoxide to H2O2, which accumulates more rapidly than it is degraded by catalase. This increase in H2O2 was inhibited by extracellular catalase, diphenylene iodonium, an inhibitor of the NADH/NADPH oxidase, and the AT1 receptor blocker losartan. In VSMCs stably transfected with antisense p22phox, a critical component of the NADH/NADPH oxidase in which oxidase activity was markedly reduced, Ang II-induced production of H2O2 was almost completely inhibited, confirming that the source of Ang II-induced H2O2 was the NADH/NADPH oxidase. Using a novel cell line that stably overexpresses catalase, we showed that this increased H2O2 is a critical step in VSMC hypertrophy, a hallmark of many vascular diseases. Inhibition of intracellular superoxide dismutase by diethylthiocarbamate (1 mmol/L) also resulted in attenuation of Ang II-induced hypertrophy (62+/-2% inhibition). These data indicate that AT1 receptor-mediated production of superoxide generated by the NADH/NADPH oxidase is followed by an increase in intracellular H2O2, suggesting a specific role for these oxygen species and scavenging systems in modifying the intracellular redox state in vascular growth. PMID- 9740617 TI - Renovascular hypertension in bradykinin B2-receptor knockout mice. AB - We evaluated whether kinins exert a protective action against the development of two-kidney, one clip (2K1C) hypertension, a model characterized by an activated renin-angiotensin system in the ischemic kidney and increased expression of the bradykinin (BK) B2 receptor in the contralateral kidney. BK B2-receptor knockout (B2-/-), wild-type (B2+/+), and heterozygous (B2+/-) mice underwent clipping of the left renal artery, with the other kidney remaining untouched. Basal systolic blood pressure (SBP, via tail-cuff plethysmography) was higher in B2-/- mice than in B2+/- or B2+/+ mice (121+/-2 versus 113+/-2 and 109+/-1 mm Hg; P<0.05 for both comparisons). SBP did not change from basal values after sham operation, but it increased in mice that underwent clipping. The increase in SBP was greater in 2K1C B2-/- mice than in B2+/- or B2+/+ mice (28+/-2 versus 14+/-2 and 14+/-2 mm Hg, respectively, at 2 weeks; P<0.05 for both comparisons). Blockade of the BK B2 receptor by Icatibant enhanced the pressure response to clipping in B2+/+ mice (29+/-2 mm Hg at 2 weeks). Intra-arterial mean blood pressure (MBP) was higher in 2K1C than in respective sham-operated mice, with the MBP difference being higher in B2-/- mice (32 and 38 mm Hg, at 2 and 4 weeks, respectively), and higher in B2+/+ mice given Icatibant (30 and 32 mm Hg) than in B2+/+ mice without Icatibant (17 and 18 mm Hg). At 4 weeks, acute injection of an angiotensin type 1 receptor antagonist normalized the MBP of 2K1C hypertensive mice. A tachycardic response was observed 1 week after clipping in B2-/- and B2+/- mice, but this effect was delayed in B2+/+ mice. However, the HR response to clipping in B2+/+ mice was enhanced by Icatibant. Within each strain, heart weight to body weight ratio was greater in 2K1C hypertensive mice than in sham-operated control animals (B2-/-: 5.7+/-0.1 versus 5.2+/-0.1; B2+/+: 5.1+/-0.1 versus 4.5+/-0.1; P<0.01 for both comparisons). The clipped kidney weight to nonclipped kidney weight ratio was consistently reduced in mice with 2K1C hypertension. Our results indicate that kinins acting on the BK B2 receptor exert a protective action against excessive blood pressure elevation during early phases of 2K1C hypertension. PMID- 9740618 TI - Association between a polymorphism in the G protein beta3 subunit gene and lower renin and elevated diastolic blood pressure levels. AB - Gi proteins mediate the intracellular effects of many vasoactive and proliferative stimuli. Recently such signaling was found to be enhanced in cultured cells of some hypertensive subjects. A polymorphism at position 825 (C- >T) of the G protein beta3 subunit gene (GNB3) was strictly related to this phenotype. The aim of the present investigation was to test the association between this polymorphism and blood pressure and plasma renin levels in humans. A population-based sample (n=608) was analyzed by questionnaire and characterized for blood pressure; plasma renin, prorenin, and aldosterone levels; and Gbeta3 C825T allele status. In individuals without antihypertensive medication (n=474; age range, 52 to 67 years), the polymorphism was mildly associated with diastolic blood pressure (CC: 88.6+/-0.3 mm Hg, n=218; versus CT: 90.1+/-0.7 mm Hg, n=209; versus TT: 91.8+/-1.7 mm Hg, n=47; P=0.02 for trend) but not with systolic blood pressure. Furthermore, the 825T allele was also significantly associated with lower renin and prorenin levels, whereas the aldosterone to renin ratio was elevated in these subjects. Significant associations between the 825T allele and diastolic blood pressure, plasma renin, and prorenin levels (inverse), and the aldosterone to renin ratio persisted after adjustment for age, gender, body mass index, and systolic blood pressure. Finally, when the entire sample was considered and an adjustment was made for covariates, the presence of arterial hypertension and the use of antihypertensive medication were both 1. 8-fold higher in the TT than in the CC genotype group (P<0.05 and P=0.06, respectively). This observation, if replicated in further studies, suggests a molecular mechanism that unifies a higher diastolic blood pressure, a lower renin level, and an elevated aldosterone to renin ratio, ie, a combination of features frequently found in patients with arterial hypertension. PMID- 9740619 TI - Differences in tissue angiotensin II-forming pathways by species and organs in vitro. AB - Angiotensin (Ang) II plays an important role in cardiovascular homeostasis, not only in the systemic circulation but also at the tissue level, and is involved in the remodeling of the heart and vasculature under pathological conditions. Although alternative Ang II-forming pathways are known to exist in various tissues, the details of such pathways remain unclear. The aim of this study was to examine tissue Ang II-forming activities and to identify the responsible enzyme in several organs (lung, heart, and aorta) in various species (human, hamster, rat, rabbit, dog, pig, and marmoset). Among the organs examined, the lung contained the highest Ang II-forming activity. The responsible enzyme for pulmonary Ang II formation was angiotensin I-converting enzyme (ACE) in all of the species except the human lung, in which a chymaselike enzyme was dominant. In the heart, the highest total Ang II-forming activity was observed in humans, and a chymaselike enzyme was dominant in all of the species except rabbit and pig. Aorta exhibited a relatively high total Ang II-forming activity, with a predominance of chymaselike activity in all of the species except rabbit and pig, in which ACE was dominant. Our results indicate that there were remarkable differences in Ang II-forming pathways among the species and organs we examined. To study the pathophysiological roles of ACE-independent Ang II formation, one should choose species and/or organs that have Ang II-forming pathways similar to those in humans. PMID- 9740620 TI - Endothelial nitric oxide synthase gene polymorphism and acute myocardial infarction. AB - Recently a point mutation of guanine to thymine at nucleotide position 1917 in the endothelial nitric oxide synthase (eNOS) gene has been reported to be associated with coronary artery spasm. In addition, a significant association of the 4a/b polymorphism in intron 4 of the eNOS gene with coronary artery disease has been reported. However, the implications of these polymorphisms with respect to acute myocardial infarction (AMI) remain to be established. We conducted a case-control study of 226 patients with AMI and 357 healthy gender- and age matched control subjects. In the former group, coronary angiograms were evaluated according to angiographic criteria based on the number of diseased vessels (>/=75%) and the number of stenotic lesions (>/=50%). Homozygosity for the Glu Asp298 polymorphism existed in 5 of 226 patients with AMI (2.2%) but not in any of the 357 control subjects (P=.0085). However, when we evaluated the coronary angiograms of 226 case patients, there was no difference in the number of diseased vessels or the number of stenotic lesions between the patients with this homozygote and those without it. By contrast, there was no evidence of a significant increase in the risk of AMI or the severity of coronary atherosclerosis among individuals with the a/a genotype of the eNOS4a/b polymorphism. Our results imply that patients who are homozygous for the Glu Asp298 polymorphism may be genetically predisposed to AMI; however, this mutation apparently is not related to the severity of coronary atherosclerosis. Further studies are needed to confirm our results and characterize the molecular mechanisms by which eNOS is involved in susceptibility to AMI. PMID- 9740622 TI - Nitric oxide synthesis inhibition retards surgical reversal of one-kidney Goldblatt hypertension in rats. AB - Surgical correction of renal artery stenosis in Goldblatt hypertension rapidly normalizes blood pressure and increases renal function. This study was conducted in 1-kidney, 1 clip (1K1C) Goldblatt hypertensive rats to examine whether the unclipping-induced reversal of blood pressure and renal function is mediated by nitric oxide (NO). The 1K1C rats were prepared and given tap water with or without supplementation of NG-nitro-L-arginine methyl ester (L-NAME). Systolic blood pressure (SBP) before and after renal artery clipping was measured with the tail-cuff method. Four weeks later, surgical unclipping was performed while blood pressure and renal function responses were determined. The results show that clipping the renal artery for 4 weeks increased SBP from 140+/-5 to 183+/-6 mm Hg (P<0.05). Concurrent L-NAME treatment accelerated and aggravated the clipping induced increases in SBP from 138+/-6 to 219+/-8 mm Hg (P<0.05). Surgical unclipping reduced blood pressure to normotensive levels within 2 hours in all hypertensive rats with and without chronic or acute L-NAME treatment. However, the magnitude of reductions in blood pressure in the initial 1 hour after unclipping was significantly less in L-NAME-treated rats than in nontreated rats (9+/-2% versus 16+/-1%, P<0.05). Despite reducing blood pressure, unclipping significantly increased glomerular filtration rate, urine flow, and sodium and potassium excretions, but the extent of the increases in these renal functions was significantly attenuated in L-NAME-treated rats. These data suggest that NO production partly contributes to the hypotensive and renal responses to unclipping but does not mediate the reversal of renovascular hypertension of this model. PMID- 9740621 TI - Regulation of angiotensin II receptor expression by nitric oxide in rat adrenal gland. AB - We recently reported that administration of Nomega-nitro-L-arginine methyl ester (L-NAME), an inhibitor of nitric oxide (NO) production, activates the vascular and cardiac renin-angiotensin systems and causes vascular thickening and myocardial hypertrophy in rats with perivascular and myocardial fibrosis. It has been reported that aldosterone may contribute to the development of cardiac fibrosis, but it is not known whether inhibition of NO synthesis affects angiotensin II (Ang II) receptor gene expression and aldosterone secretion. The aim of this study was to investigate the effect of NO inhibition on the expression of Ang II receptors in the adrenal gland and on aldosterone secretion in rats. Wistar King A rats received normal water, L-NAME alone (1 mg/mL in the drinking water), or L-NAME and the alpha1-adrenergic receptor blocker bunazosin (0.1 mg/mL in the drinking water) for 1 week. After 1 week of treatment with L NAME, systolic blood pressure, plasma aldosterone concentration (PAC), and mRNA level and number of Ang II type 1 receptor (AT1-R) were increased. Plasma renin activity, serum angiotensin-converting enzyme activity, and the number of AT2-R were unchanged. Although addition of bunazosin to L-NAME restored systolic blood pressure to the control level, PAC and AT1-R numbers remained significantly higher than those of control level. These results suggest that the increased AT1 R number and PAC induced by the inhibition of NO synthesis were independent of blood pressure and systemic renin-angiotensin system. Therefore, hypertension and myocardial fibrosis induced by NO blockade may be due in part to an elevation of PAC caused by increased AT1-R in the adrenal gland. PMID- 9740623 TI - Acute endothelium-mediated vasodilation is not impaired at the onset of diabetes. AB - Vascular injury and impaired vascular function are central to the increased mortality associated with diabetes. Hyperglycemia in diabetes has been suggested to play a role in this process, in part by impairing the function of the vascular endothelium. It has been difficult, however, to isolate the direct effect of glucose in both humans and in animal models of diabetes. This was evaluated in the present study in 7 rats that were chronically instrumented with a Transonic flow probe at the iliac bifurcation of the abdominal aorta, a nonoccluding catheter inserted immediately anterior to the flow probe, and a femoral vein catheter. Acute infusions of acetylcholine and sodium nitroprusside (1 and 10 microg/min IA) increased hindquarter blood flow significantly by approximately 27 and 10 mL/min over baseline, respectively, at the high dose. Streptozotocin (70 mg/kg IV) was administered, but normoglycemia was maintained with continuous intravenous insulin infusion to control for potential streptozotocin side effects. Diabetes was induced 5 to 7 days later by stopping the insulin infusion. Hindlimb blood flow (measured 24 hours per day) decreased during the diabetic period and was accompanied by an increase in mean arterial pressure, suggesting a vasoconstrictor response. However, the responses to acetylcholine and sodium nitroprusside were not altered significantly on either day 2 or day 6 of the diabetic period. This suggests that neither endothelium-mediated vasorelaxation nor responsiveness to nitric oxide is impaired during the initial phase of diabetes and that diabetic hyperglycemia does not have a significant, direct effect to impair endothelium-mediated relaxation in insulin-dependent diabetes mellitus. The mechanism for the change in baseline blood flow and its potential influence on endothelial function, however, are not known. PMID- 9740625 TI - Determinants of pulse pressure. AB - We have searched to define the major arterial parameters that determine aortic systolic (Ps) and diastolic (Pd) pressure in the dog. Measured aortic flows were used as input to the 2-element windkessel model of the arterial system, with peripheral resistance calculated as mean pressure divided by mean flow and total arterial compliance calculated from the decay time in diastole. The windkessel model yielded an aortic pressure wave from which we obtained the predicted systolic (Ps,wk) and diastolic (Pd,wk) pressures. These predicted pressures were compared with the measured systolic and diastolic pressures. The measurements and calculations were performed for 7 dogs under control conditions during aortic occlusion at 4 locations (the trifurcation, between the trifurcation and diaphragm, the diaphragm, and the proximal descending thoracic aorta) and during occlusion of both carotid arteries. Under all conditions studied, the predicted systolic and diastolic pressures matched the experimental ones very well: Ps,wk=(1.000+/-0.0055) Ps with r=0.958 and Pd,wk=(1.024+/-0.0035) Pd with r=0.995. Linear regression for pulse pressure (PP) resulted in PPwk=(0.99+/ 0.016) PP with r=0.911. We found the accuracy of prediction equally good under control conditions and in the presence of aortic or carotid artery occlusion. Multiple regression between pulse pressure and arterial resistance and total arterial compliance yielded a poor regression constant (R2=0.19), suggesting that the 2 arterial parameters alone cannot explain pulse pressure and that flow is an important determinant as well. We conclude that for a given ejection pattern (aortic flow), 2 arterial parameters, total arterial resistance and total arterial compliance, are sufficient to accurately describe systolic and diastolic aortic pressure. PMID- 9740624 TI - ANP and bradycardic reflexes in hypertensive rats: influence of cardiac hypertrophy. AB - In previous studies we demonstrated that in normotensive rats, but not in spontaneously hypertensive rats (SHR), atrial natriuretic peptide (ANP) enhances bradycardic reflexes through an action on cardiac vagal afferent pathways. The present study aimed to determine whether cardiac hypertrophy, hypertension, or a nonreversible genetic factor accounted for the insensitivity of SHR to ANP action on cardiac reflex pathways. SHR were treated with the angiotensin-converting enzyme (ACE) inhibitor perindopril (3 mg/kg per day) for 6 weeks from 4 to 9 weeks of age (SHR-S, n=10) or for 9 weeks from 4 to 12 weeks of age (SHR-L, n=10) or were untreated (SHR, n=10) to produce differential effects on blood pressure and left ventricle/body weight ratio (LV/BW). Untreated normotensive Wistar-Kyoto rats (WKY, n=10) were also studied. At 13 weeks of age, all rats were instrumented with aortic and jugular catheters, and at 14 weeks we measured heart rate reflexes to rapid intravenous infusions of methoxamine (100 microg/kg, cardiac baroreflex) and serotonin (5 to 60 microg/kg, von Bezold-Jarisch cardiac chemosensitive reflex), with either alpha-rat ANP (150 ng/kg per minute IV) or saline vehicle (270 microL/h IV) infusion. Perindopril treatment for 6-week (SHR S) and 9-week (SHR-L) durations maintained blood pressure at normotensive levels in both groups. SHR-S exhibited a small degree of cardiac hypertrophy (LV/BW was 8% higher than in WKY but 11% less than in untreated SHR), but LV/BW was normalized in SHR-L (to within 1% of WKY LV/BW). In WKY, ANP significantly (P<0.05) enhanced bradycardic responses to both the cardiac baroreflex (by 42+/ 10%) and von Bezold-Jarisch chemosensitive reflex (by 17+/-5%) activation but had no effect in SHR. The cardiac reflex action of ANP was restored in SHR-L (ANP enhanced reflex bradycardia by 28+/-12% and 36+/-8%, baroreflex and von Bezold Jarisch reflex, respectively; P<0.05), but SHR-S, which developed some cardiac hypertrophy, remained unresponsive to ANP. Our results suggest that the inability of ANP to sensitize cardiac vagal (nonarterial) afferents in SHR was not due to an inherited irreversible component, or the hypertension per se, but was associated with the presence of cardiac hypertrophy. A functional consequence of hypertension-induced cardiac hypertrophy may be the inhibition of the cardioprotective action of ANP through cardiac vagal reflexes. PMID- 9740626 TI - Pulse pressure and cardiovascular mortality in normotensive and hypertensive subjects. AB - There is now increasing evidence that high pulse pressure, which is an indicator of large artery stiffness, is an independent risk factor for cardiovascular mortality, especially coronary mortality, in different populations. We have recently shown in a large French population that in male subjects aged 40 to 69 years, increased pulse pressure was a strong predictor of cardiovascular mortality, especially coronary mortality. In the present report, we analyzed the effect of pulse pressure in men and women of the same cohort after classifying them as normotensive (systolic blood pressure [SBP] <140 mm Hg and DBP <90 mm Hg) or hypertensive (SBP >/=160 mm Hg or DBP >/=95 mm Hg). After adjustment for age, mean blood pressure, and other risk factors, the relative risk (95% confidence limits) for cardiovascular mortality for an increase of 10 mm Hg of pulse pressure was 1.20 (1.01 to 1.44) in normotensives and 1.09 (1.03 to 1.14) in hypertensives. Cardiovascular and coronary death rates were similar in the group of normotensive men with a pulse pressure >50 mm Hg and in the group of hypertensive men with a pulse pressure <45 mm Hg. No association between cardiovascular mortality and pulse pressure was observed in either normotensive or hypertensive women (0.85 [0.60 to 1.21] and 1.0 [0. 91 to 1.11], respectively). Low mortality rates could explain this observation in normotensive but not in hypertensive women, in whom cardiovascular mortality rates were relatively high. Because a high pulse pressure in men is an independent predictor of cardiovascular mortality in both hypertensives and in those considered as having normal blood pressure, this parameter could aid in evaluating cardiovascular risk. PMID- 9740627 TI - Relation between number of cardiovascular risk factors/events and noninvasive Doppler ultrasound assessments of aortic compliance. AB - The aim of this study was to establish the relation between noninvasive Doppler ultrasound assessments of aortic compliance, based on "foot-to-foot" aortic pulse wave velocity measurements, and presumed atherosclerotic load in patients with vascular disease and/or diabetes mellitus. One hundred ten patients with vascular disease and/or diabetes mellitus (arteriopaths) underwent measurement of in vivo aortic compliance using Doppler ultrasound. Demographic data on these subjects were recorded along with details of cardiovascular risk factors and events. Aortic compliance values were compared with data from 51 age-matched healthy, asymptomatic subjects putatively free of vascular disease (controls). Data are expressed as mean+/-SD. Arteriopaths were aged 64.1+/-8.4 years and had total cholesterol levels of 5.9+/-1.1 mmol/L and aortic compliance of 0.78+/-0.42%/10 mm Hg [1.33 kPa]. Most arteriopaths had 2 or more cardiovascular risk factors and events: diabetes (n=41), hypertension (n=45), smoking (n=86), cerebrovascular/transient ischemic event (n=13), myocardial infarction (n=44), angina (n=51), and/or peripheral vascular disease (n=33). Controls were aged 64.3+/-12.1 years with total cholesterol of 6.1+/-1.1 mmol/L and aortic compliance of 1.14+/-0.46%/10 mm Hg [1.33 kPa] (P<0.002 versus arteriopaths). Subset analysis revealed that patients with the greatest number of cardiovascular risk factors and events (n=5) had the stiffest aortas (aortic compliance, 0.58+/ 0.15%/10 mm Hg [1.33 kPa]) compared with those patients with the median and mean (n=2) number of risk factors and events (aortic compliance, 0.80+/-0.50%/10 mm Hg [1.33 kPa]; P<0.02). The data suggest that a significant inverse relation exists between presumed atherosclerotic load (as assessed by the number of cardiovascular risk factors and events) and aortic compliance determined noninvasively based on aortic pulse wave velocity measurements. If these findings are confirmed by prospective, longitudinal follow-up studies, such measurements may prove useful as a noninvasive marker of vascular risk. PMID- 9740628 TI - Carotid arterial stiffness as a predictor of cardiovascular and all-cause mortality in end-stage renal disease. AB - Damage of large arteries is a major contributory factor to the high pulse pressure observed in patients with end-stage renal disease. Whether incremental modulus of elasticity (Einc), a classic marker of arterial stiffness, can predict cardiovascular mortality has never been investigated. A cohort of 79 patients with end-stage renal disease undergoing hemodialysis was studied between September 1995 and January 1998. Mean age at entry was 58+/-15 years. The duration of follow-up was 25+/-7 months, during which 10 cardiovascular and 8 noncardiovascular fatal events occurred. At entry, carotid Einc was calculated from measurements of diameter, thickness (echo-tracking technique), and pulse pressure (tonometry). Based on Cox analyses, 2 dominant factors emerged as predictors of all-cause and cardiovascular mortality: increased Einc and decreased diastolic blood pressure. Lipid abnormalities and the presence of previous cardiovascular events interfered to a smaller extent. After adjustment for confounding variables, the odds ratio for Einc >/=1 kPa-3 was 9.2 (95% confidence interval, 2.4 to 35.0) for all-cause mortality. These results provide the first direct evidence that in patients with end-stage renal disease undergoing hemodialysis, arterial alterations, as determined from carotid Einc, are strong independent predictors of all-cause and cardiovascular mortality. PMID- 9740629 TI - Relation between coronary artery disease, aortic stiffness, and left ventricular structure in a population sample. AB - To elucidate the relationship between coronary artery disease (CAD), aortic stiffness, and left ventricular structure, we recruited 55 subjects (33 men; average age, 63+/-1 years) with previously unknown CAD from a healthy general population sample, as well as 55 control subjects matched for gender, age, and serum cholesterol level. We measured arterial blood pressure and the systolic expansion of the transverse aorta and left ventricular structure by echocardiography. Aortic stiffness was higher in CAD patients than in controls, with a brachial pulse pressure of 59+/-3 versus 52+/-2 mm Hg and stiffness indices of Ep=212+/-26 versus 123+/-13 kN/m2 and beat=16+/-2 versus 9+/-1 (all P<0.01). Mean arterial pressure was similar in both groups during the measurements (95+/-2 versus 93+/-2 mm Hg, P=NS). Most CAD patients (61%) were in the highest stiffness quartile defined by the normal control values (P<0.05 versus control). Left ventricular mass index was also higher in CAD patients than in matched controls (139+/-5 versus 123+/-4 g/m2, P<0.05). We conclude that aortic stiffness and left ventricular mass are increased in subjects newly diagnosed as having CAD. This might explain previously reported associations of an increased mortality, particularly from CAD, found among subjects with elevated pulse pressures. PMID- 9740630 TI - Lack of association between renin-angiotensin system, gene polymorphisms, and wall thickness of the radial and carotid arteries. AB - To investigate the relationship between polymorphisms of the angiotensin converting enzyme (ACE) and the angiotensin II type 1 receptor (AT1R) genes and structural phenotypes of arteries, we studied a cohort of 340 subjects (aged 49+/ 12 years) without evidence of cardiovascular disease and who had never been treated previously with any cardiovascular treatments. Structural phenotypes (wall thickness and internal diameter) were evaluated for the common carotid and the radial arteries using high-resolution echo-tracking devices (NIUS-02 and Wall Track System). The influence of ACE insertion/deletion (I/D) and AT1R A/C1166 polymorphism genotypes on structural parameters was tested by ANOVA and logistic regression analysis. For the radial artery, mean wall thickness among subjects according to the ACE I/D or AT1R A/C1166 genotypes was not different. This lack of association persisted in a logistic regression analysis or when the comparison was restricted to a subgroup of subjects potentially at high genetic risk (DD and CC or AC) compared with subjects at low genetic risk (AA and II or ID). Also, no association was observed between the carotid artery intima-media thickness and the 2 polymorphisms. In conclusion, the ACE I/D and the AT1R A/C1166 gene polymorphisms are not markers of vascular hypertrophy in subjects with no evidence of cardiovascular disease. These results suggest that these gene polymorphisms have an undetectable role in the geometry of the radial and carotid arteries compared with usual determinants such as blood pressure and age. PMID- 9740632 TI - AHA journals at the forefront of cardiology : A report on excellence PMID- 9740631 TI - Effects of prolonged immobilization of the limb on radial artery mechanical properties. AB - Physical training is associated with an increase in arterial distensibility. Whether the effect of training on this variable is evident also for ordinary levels of exercise or no exercise is unknown, however. We have addressed this issue by investigating the effect on radial artery distensibility of prolonged monolateral immobilization of the ipsilateral limb versus the following resumption of normal mobility. We studied 7 normotensive subjects (age, 25.4+/ 3.0 years; systolic/diastolic blood pressure, 119+/-9/68+/-6 mm Hg, mean+/-SE) in whom 1 limb had been immobilized for 30 days in plaster because of a fracture of the elbow. At both the day after plaster removal and after 45 days of rehabilitation, radial artery distensibility was evaluated by an echo-tracking device (NIUS-02), which allows arterial diameter to be measured noninvasively and continuously over all pressures from diastole to systole (finger monitoring), with the distensibility values being continuously derived from the Langewouters formula. In both instances, the contralateral arm was used as control. Immediately after removal of the plaster, radial artery distensibility was markedly less in the previously immobilized and fractured limb compared with the contralateral limb (0.4+/-0.1 versus 0.8+/-0.1, 1/mm Hg 10(-3), P<0.05). After rehabilitation, the distensibility of the radial artery was markedly increased in the previously fractured limb (0.65+/-0.1 1/mm Hg 10(-3), P<0.05), whereas no change was seen in the contralateral limb. Thus, complete interruption of physical activity is associated with a marked reduction of arterial distensibility, indicating that even an ordinary level of activity plays a major role in modulation of arterial mechanical properties. PMID- 9740633 TI - New priorities for health sector reform in Central and Eastern Europe. AB - After the breakdown of the state socialism, a number of changes have occurred in the legal framework, as well as governmental policy, ownership, production, financing, and reimbursement of health care in Central and Eastern Europe (CEE). However, the policy context in CEE makes priority setting a necessary step to ensure the efficient use of public funds for health. The problems with prioritizing of health services in the Central and Eastern Europe are, in essence, related to the general position of health care within broad national priorities. The percentage of gross domestic product spent on health is insufficient and many cost-effective interventions are currently neglected, under funded or provided with low quality standards. If the health status is to be improved, such interventions should be granted a greater priority. The experience from the established market economies indicate that: (a) overall future system of priorities setting in health care in the CEE should be driven by new democratic values; (b) new systems must be people-centered, more oriented to the needs of individual patient and specific groups, and sensitive to inequalities, unemployment, and social poverty; (c) they should be health-focused; and (d) they should be evidence-based and oriented towards primary health care. PMID- 9740634 TI - Managing physician resources: East and West. AB - AIM: To examine the current physician supply in western countries, Central and Eastern Europe (CEE), and the Newly Independent States (NIS) of the former Soviet Union. To assess the current management of physician resources in these regions and the potential for their future management. METHODS: Description and analysis of current physician to population ratios, graduating physician to population ratios, and specialty distribution of physicians. RESULTS: Maldistribution of physician resources, both by specialty and geographically, exists in the East and the West. The management of physician resources varies widely in the West, from virtually no attempts to manage the resources in the United States to increasingly regulatory methods in Canada. The CEE and the NIS face problems in managing physician resources as the movement to primary care continues. At the same time, changes in payment mechanisms create new and often perverse incentives for physicians, in addition to the problems inherited from the centrally managed systems. CONCLUSIONS: Countries in the East and West face challenges in managing physician resources to overcome the current maldistribution of those resources. Efforts must be made to make the medical education system responsive to the future physician supply needs. This is especially true in the East where health systems continue the transition from centrally managed to more market based systems. PMID- 9740635 TI - Is public health between East and West? Analysis of wealth, health and mortality in Austria, Central and Eastern European Countries and Croatia relative to the European Union. AB - AIM: To provide a conceptual framework for health planning activities in the "middle income" transition countries. METHOD: Economic, demographic, and disease related data in Central and Eastern European (CEE) countries, including Croatia and Austria, were compared to the Europen Union (EU) average. Data were selected from the databases provided by the World Health Organization, Organization for Economic Cooperation and Development, World Bank, United Nations, and the European Bank of Reconstruction and Development. Life expectancy and mortality were extrapolated until the year 2000 by using an exponential growth model for the WHO time series data, starting in 1994. Death rates due to ischemic heart diseases (18%) and cerebrovascular diseases (13%) were selected to show frequent causes of death. RESULTS: Relative to the EU average, the gross domestic product (GDP) share of health expenditures in transition countries was disproportionate to wealth and premature death. The population in CEE-countries was younger and the share of people aged >65 was predicted to remain about 15% below the EU average and Austria. For Croatia, the share of people aged 65 would be on the increase, similar to the share predicted for Austria (slightly above the EU average). Mortality of selected non-communicable, chronic diseases is predicted to increase and remain relatively high. Mortality rates due to infectious diseases have been declining but remained comparatively on a high level. CONCLUSIONS: Coexistence of demographic and epidemiological transition along with high mortality rates due to infectious diseases creates a "double burden". Economic transition has the potential to comprise both the increase in wealth, and life and health expectancy. PMID- 9740636 TI - Organization of health care in Croatia: needs and priorities. AB - AIM: Description and analysis of the present situation of health care system in Croatia, and its characteristics in the transitional process of restructuring. METHODS: A descriptive method was used. The data from the regular statistical publications were used for the analysis. RESULTS: Croatia is faced with problems similar to those in other countries of the Central and Eastern Europe (CEE), such as control of health expenditure, balancing the development of different segments of health care services, stabilization of effectiveness and quality of care, transition from one-party system to pluralistic democracy, introduction of a free market economy, war devastations, etc. On the other hand, the Croatian experience in the development of a decentralized and integrated primary health care, decentralized health insurance system, education of general/family practitioners, and a tradition in the implementation and development of public health measures, have facilitated and contributed positively to the whole process of transition. CONCLUSION: In contrast to the economic difficulties, war devastations, and changing the social system, the Croatian health care system proved its stability and sustainability. The highest priority and needs are now related to coping with unhealthy behavior of the population, such as smoking, accidents, physical inactivity, and nutritional problems, which should be solved and controlled by the implementation of preventive programs, organization and management of public health services, and further focusing onto the integrated type of primary health care in the organization of services. Hospital services need more intensive and skilled management, as well as support measures for better quality of work. PMID- 9740637 TI - Health care reform in Croatia: the consumers' perspective. AB - AIM: Assessment of the Croatian health care system (under the reform) from the perspective of the users of health care services. We analyzed the consumers' satisfaction with health care system, health care expenses and access, and described the consumers' attitudes toward health reform, examining the differences among sociodemographic groups. METHODS: The study is based on a data set collected in 1994 through the interviews with randomly selected adults in two major cities of Croatia: Zagreb and Split. RESULTS: A great proportion of respondents were dissatisfied with the current health care services, quality of health care facilities and equipment, and encountered difficulties in access. The elderly, women, and those with lower socioeconomic status were more likely to be dissatisfied and to consider out-of-pocket payments for health services as a problem. A great number of the respondents believed that the reform would either fail or would not achieve significant results. Compared to the younger and higher socio-economic group, the older and lower socioeconomic groups were more likely to evaluate the health care reform negatively. CONCLUSION: Croatian government decided to rationalize the health care system without taking much account of the impact of health reform on the consumers. Revealed dissatisfaction with the health care services might be linked with the expressed doubts in health care reform and concern that changes could worsen the consumers' position as patients. PMID- 9740638 TI - Application of the modified method of "rapid appraisal to assess community health needs" for making rapid city health profiles and city action plans for health. AB - AIM: To develop a method that Croatian cities could use in the development of the City Health Profile and City Health Plan. The assessment concerned cities that have recently experienced the war and thus the method had to be rapid, cheap, scientifically based, sensitive, participative (involving politicians, experts, and citizens), able to produce immediate action, and to sustain the gained benefits. METHOD: A utilization-focused strategy was selected. Through ongoing interactions with intended information users, research questions were focused and the method of Rapid Appraisal to assess community health needs was selected as appropriate. This method was modified to: 1. assess the health of each city and serve as the basis for creating the City Health Profile; 2. select (Healthy City Project) priority areas; 3. establish the working groups on priority areas; and 4. build on the three previous steps to develop the City Action Plan for Health. RESULTS: During 1996, the Rapid Appraisal was applied in three Croatian cities (Pula, Metkovic, Rijeka). The work resulted in the completion of the City Health Profile, selection of the Project Priority Areas, formation of thematic working groups on priority areas, and acceptance of the agreed City Action Plan for Health. The method provided a scientifically based account of health in each of the three cities and identified targets for the future by using health-related measures and citizens' observations about the community, its problems, and potentials. CONCLUSION: The method proved to be credible and sensitive to the social and cultural differences it encompassed. PMID- 9740639 TI - Priority setting and scarce resources: case of the Federation of Bosnia and Herzegovina. AB - The priority setting within the context of scarce resources in the Federation of Bosnia and Herzegovina (BH) can be divided into priorities within the health care services provision and priorities within the reconstruction process. Facing the resource scarcity, the Federation of BH has chosen to increase health insurance contribution rate to establish cost-sharing arrangements (co-payments) and priority setting to ration access to certain services funded by the compulsory health insurance ("basic package" of the health care services). Reconstruction process of the health care facilities is conducted on the federal level through the effective managerial infrastructure ("project implementation units"). This study reports on the consequences of the war in BH as a peculiar context of the overall reform objectives and priority setting. PMID- 9740640 TI - Rebuilding the healthcare system in Mostar: challenge and opportunity. AB - A basic premise of this article is that the political and ethnic division of the city of Mostar imperils community health planning efforts beyond those normally encountered in the aftermath of war. Bosnia and Herzegovina is divided among ethnic groups, and the city of Mostar is further subdivided between the Croatian and Muslim interest groups. In that respect, Mostar presents a very special challenge to those planning the reconstruction of the health care delivery infrastructure. A new healthcare delivery plan was organized by the Federation of Bosnia and Herzegovina in 1996. This paper examines obstacles involved in implementing that plan in light of some epidemiological indicators of current health conditions. PMID- 9740641 TI - Priorities for medium-term development in the health sector of Bosnia and Herzegovina. AB - In the medium term, the World Bank will work with the Government of Bosnia and Herzegovina, non-government organizations, and international development agencies to complete the reconstruction work already begun. There will be an increased emphasis on developing sustainable health finance regime and establishing a basic health system. PMID- 9740642 TI - Privatization of health care in Slovenia. AB - This article analyzes the results of the hitherto privatization of health care in Slovenia based on statistical indices and public opinion data. The new legislation offered much more room for quasiprivatization than for real privatization. A greater room was left for privatization of financing than for the status of health care providers and infrastructure. The statistical data on private practitioners and the share of private resources in health care costs show that the privatization is relatively slow but irreversible process. Public opinion data revealed that the population of Slovenia accepted this process but some signs of distrust in private practice and new forms of health insurance were noticed. The new legislation leaves a high degree of regulation to the state in protecting citizens' rights but there are some risks of unrestrained development of the market due to insufficient implementation of legislation. PMID- 9740643 TI - Priorities of the Russian health care reform. AB - The introduction of health insurance system has been the core of the Russian health care reform. It has coincided with the decentralization of the state administration. The reform has thus been decentralized, and the transition has been fragmentary and incomplete. As a result, the existing health financing system is eclectic and contradictory. Meanwhile, the reform has had a positive stabilizing influence on financing of health care under conditions of continued economic crisis. The new priorities of the reform should be to balance the financial flows and the state's obligations, and to increase the efficiency of the use of resources through encouragement of competition, assurance of transparency of public funding, development of health care planning, and shift from inpatient to outpatient care. PMID- 9740644 TI - Old priorities and new agenda of public health in India: is there a mismatch? AB - AIM: Structural adjustment programs and health sector reforms in India create new agenda in health services. The aim of this study is the analysis of the mismatch between the new health services agenda and epidemiological priorities, and exploration of the possible alternatives, such as decentralization. METHOD: Analysis of the disease pattern, health, and demographic indicators versus the pattern of investments and policy changes was performed. The possible alternatives were explored by the study of decentralization process attempted in Kerala, by anthropological field work and examination of primary documents. RESULTS: The post-reform period has witnessed a rise in the cases of communicable diseases and an increase in the frequency of epidemics in different parts of the country. The infant mortality rate has risen or decline has slowed down in many states. The new agenda, which includes gradual withdrawal of the State in the provision of health services and the process of privatization, has already been initiated in the country as a part of health sector reforms. The process of decentralization in Kerala was examined to highlight the role of people's planning in health under growing conditions of mismatch between the epidemiological priorities and new agenda. CONCLUSIONS: The new agenda could pose a serious threat to effective implementation of public health programs including control of communicable diseases. Public health programs and allocation of resources need to be rooted in the epidemiological and social context. The process of decentralization attempted in Kerala could be used as a tool to achieve this aim. PMID- 9740645 TI - Health care system of the United States and its priorities: history and implications for other countries. AB - The health care system of the United States is examined from the end of the 19th century to the present, using secondary sources on labor and health care. During that period, several actors, each with its own priorities, exercised control over the United States (US) health services: physicians (from the 1900s on), hospitals and not-for-profit insurance (from the 1930s on), governmental regulators (from the 1960s on), and, lastly, for-profit managed care enterprises (from the 1980s on). A class contest between corporations and labor was involved at two critical points. In the 1870s to 1890s (with further steps in the 1920s and, with the Taft Hartley law, in 1947), it weakened the labor movement that was unable to mount a successful effort for a national health program in 1972 and 1992. In the 1980s and 1990s, as health services developed into a major industry, two contending business groups (health plans and payers) took commanding positions over consumers and employees. Market-oriented, for-profit managed care organizations came to play a dominant role. During that period, access to, and, by some measures, quality of care has declined. The rise in health care costs has been interrupted, but it is not clear how long this will last. European nations that are reforming their health care system, should be wary of such profit-oriented market approaches to bring costs down. PMID- 9740646 TI - Health targets as an instrument for improving the rational framework of healthcare decisions. AB - There is currently a trend gaining acceptance that explicitly recognizes the need to set priorities for making rational decisions in health policy, respecting at the same time the underlying purpose of health care - to improve people's health. This trend in health policy is referred to as "health targets" although definitions vary considerably. Having been initiated by the World Health Organization in the late 1970s, the international policy on health targets is in the process of renewal to become Health for All in the Twenty-First Century. The new program highlights 10 global targets, from the reduction of worldwide burden of diseases to improvement of access to comprehensive, essential quality care. Countries such as the United Kingdom and Australia have adopted and implemented such programs which basically include cardiovascular, cancer, accidents, and mental health targets. For many countries, however, there are several difficulties in establishing similar programs. One of them is the unavailability of reliable data, although political factors and structure of health systems also play a key role in the creation of health targets. Overall, health targets appear to be a key element in building a strong public health policy, taking into consideration not only the cost of healthcare, but also the outcomes in improving health which is the ultimate goal of health care systems. PMID- 9740647 TI - Rationing of hospital services in the Australian health system. AB - This article reports on the rationing in the Australian hospital sector and explains why it has been undertaken. It also briefly overviews the Australian health system in order to provide a necessary background for the issue of rationing itself. Rationing of hospital services has occurred because governments in Australia have limited hospital sector resources trying to ensure the containment of their health budgets. The resources available to hospitals have been insufficient to ensure that the supply of services meets the demand for such services. Therefore, in order to contain hospital budgets rationing has been required. Medicare, the universal health insurance system, assures that access to public hospital services is on the basis of clinical needs. However, due to the federal nature of government in Australia, the available services are determined by health system structural interrelationships and direct government regulation. For example, services provided in the community sector, and funded by the Commonwealth government, are prime candidates for being removed from the hospital sector by State/Territory governments. Similarly, expensive services with a wide range of usage are candidates for regulation to contain costs. PMID- 9740648 TI - Priorities and priority-setting in health care in the Netherlands. AB - Since 1990, the priority-setting has become one of the key issues in making choices in health care. In 1991, the now famous Dunning Report was presented to the Dutch Cabinet. One of its main conclusions was that health services should satisfy four criteria: necessary care, effectiveness, efficiency, and individual responsibility. Priority-setting can be done either by excluding medical treatments from compulsory health insurance coverage and/or by the use of both protocols and guidelines, and the individual selection of patients by health professionals. The discussion on the introduction of in vitro fertilization into the basic health insurance package and the exclusion of dental care for adults have shown that, on the basis of the Dunning criteria, it is not easy to leave complete or parts of services out of the basic health insurance package. The second strategy - the application of the Committee's criteria by the use of protocols, guidelines, and budget restrictions - is even more difficult to realize. More patients assert their right to health care benefits before courts. The courts' decisions have shown that it is difficult for the patient's counsellor to prove that government is responsible for non-delivery due to force majeur. Courts attach much importance to the Dunning criteria; in particular the criterion of necessity. PMID- 9740649 TI - Health status of refugees from former Yugoslavia: descriptive study of the refugees in the Netherlands. AB - AIM: To describe the health status of formally recognized refugees from the former Yugoslavia in the Netherlands, and analyze the relationship between experienced traumatic events and health status. METHODS: A random sample of refugees was taken from randomly selected municipalities. One hundred and two persons received a postal questionnaire. Forty percent filled out the questionnaire completely. Well-validated self-assessment scales were used to measure six dimensions of health status. Traumatic experiences were assessed by the Harvard Trauma Questionnaire. Physical aspects of health status were measured by the three dimensions of the Medical Outcome Study (MOS) Short-form General Health Survey (SF-20) which consisted of the dimensions "physical functioning", "subjective health", and "pain". Social health was measured by the two dimensions of the SF-20, "role fulfilling" and "social functioning". RESULTS: The refugees showed vulnerable health. A significant group had an accumulation of health problems. These were mostly unrelated to sociodemographic variables. Many of the refugees experienced several traumatic events. These experiences were clearly related to all health problems. CONCLUSIONS: The refugees from the former Yugoslavia scored low on standardized health measurement scales. This was primarily due to their traumatic experiences in combination with their refugee status. There is a need for health services to prevent the accumulation of (future) health problems. PMID- 9740650 TI - Five-year review of diarrheal disease cases admitted to a busy referral hospital in Ghana. AB - AIM: To evaluate successes in the clinical management of severe diarrheal diseases in a busy referral hospital in Ghana, four years after the introduction of the World Health Organization's protocol for the clinical management of diarrhea and the establishment of an oral rehydration therapy Corner. METHOD: Data on the cases of diarrheal diseases recorded in the hospital from 1992 to 1996 were collected and analyzed. RESULTS: The average overall diarrheal disease mortality over the period was around 20% with twice as much deaths among adults than among children. There was a tendency of decline in childhood mortality, whereas it was much less evident among the adults. The high mortality caused by diarrheal diseases in the hospital and the differences in adult and childhood mortality were related to the problems in case management that stemmed from diarrhea case management training of clinical staff with a bias towards the pediatric staff, and also from the loss of several trained staff members through transfers and other staff replacements within the hospital. There were similarities in the trend of admissions for adults and children over the period, which suggested a possible common etiology for severe diarrheal diseases recorded in the hospital. CONCLUSIONS: Diarrheal mortality in a busy referral hospital should be investigated regularly for lapses in management because some of these deaths may be prevented by simple interventions. PMID- 9740651 TI - Care dependency and survival among female patients with Alzheimer's disease: a two-year follow-up. AB - AIM: To investigate the relationship between the severity of the patient's care dependency on the one hand, and causes of death, co-morbidity, and survival on the other, and to find out which of these factors might be longitudinal predictors of survival. METHODS: A two-year follow-up study started in 1994 in the Netherlands. Subjects were 130 female nursing-home patients with Alzheimer's disease (91 severely dependent, and 39 mildly dependent). Features analyzed in 1994 included the scale of care dependency, demographic data, and clinical information. RESULTS: In 1994, both subsamples differed significantly with respect to the duration of Alzheimer's disease, duration of stay, and all nursing care dependency features, but they did not differ significantly in age, age of disease onset, and co-morbidity. In both groups, the main causes of death were cachexia and/or dehydration. Survival prognosis varied with the level of care dependency. Compared to mildly dependent patients, patients with severe dependency had a 20% higher mortality than expected for the general population of the same age. Marital status, education, cardiovascular disease, and four care dependency features: body posture, day/night pattern, communication, and contact with others, are factors that significantly predict survival. CONCLUSION: Survival prognosis of patients with Alzheimer's disease varies with the level of care dependency. PMID- 9740652 TI - Doctor-patient relationship in oncological illness: the "talking medicine". AB - For any physician, sympathetic interaction with his/her patients should remain a central concern. It is his/her task to understand the patient's hopes, fears, anxieties and social situation, as well as to understand himself and his own motives and attitudes, which can often be identified as helplessness. Continually advancing technology, rationalization, time, and the pressure for success leave less and less room for such considerations in medical practice. While patients often consider their medical care as very good, they, however, complain that the emotional support is often insufficient. Thus, for the benefit of both the doctor and the patient, every physician has to assure compassion in his relationship with the patients. Medical doctors taking care of cancer patients do not need only thorough medical training but also additional training in psychosomatic medicine. PMID- 9740653 TI - FGF-10 is a chemotactic factor for distal epithelial buds during lung development. AB - Fibroblast growth factor (FGF) signaling is required for normal epithelial branching in the respiratory system of several species. Recent studies have shown that FGF-10 may be a key regulator of lung branching morphogenesis, based on its pattern of expression in the early lung and its ability to induce epithelial budding in vitro. In this study we investigate whether FGF-10 is able to direct lung epithelial buds to proper positions during development . We maintained localized high levels of FGF-10 in cultured lungs using FGF-10-soaked heparin beads. FGF-10 exerts a powerful chemoattractant effect on the distal but not on proximal lung epithelium. Epithelial buds grow toward an FGF-10 source within 24 h, and subsequently form concentric layers of epithelium around the bead. BrdU incorporation analysis suggests that FGF-10, in contrast to FGF-7, is a modest proliferation factor for the lung epithelium. In the absence of mesenchyme FGF-10 requires an associated proliferative signal to induce bud migration. This can be provided by extract from lung mesenchyme, or by FGF-7, a growth factor also present in the early embryonic lung. FGF-10 does not seem to interfere with early epithelial cell differentiation. The chemoattractant effect of FGF-10 in the lung epithelium is reminiscent of the patterning effect of the Drosophila FGF ortholog branchless in the developing tracheal epithelium, suggesting that the function of these genes has been conserved during evolution. PMID- 9740654 TI - Centromeric protein B null mice are viable with no apparent abnormalities. AB - The centromere protein B (CENP-B) is a centromeric DNA/binding protein. It recognizes a 17-bp sequence motif called the CENP-B box, which is found in the centromeric region of most chromosomes. It binds DNA through its amino terminus and dimerizes through its carboxy terminus. CENP-B protein has been proposed to perform a vital role in organizing chromatin structures at centromeres. However, other evidence does not agree with this view. For example, CENP-B is found at inactive centromeres on stable dicentric chromosomes, and also mitotically stable chromosomes lacking alpha-satellite DNA have been reported. To address the biological function of CENP-B, we generated mouse null mutants of CENP-B by homologous recombination. Mice lacking CENP-B were viable and fertile, indicating that mice without CENP-B undergo normal somatic and germline development. Thus, both mitosis and meiosis are able to proceed normally in the absence of CENP-B. PMID- 9740656 TI - Mannose-specific recognition mediates two aspects of synaptic growth of leech sensory afferents: collateral branching and proliferation of synaptic vesicle clusters. AB - The developmental role of carbohydrate markers in the genesis of neuronal networks was studied using leech sensory afferents as a model. Leech sensory afferents express a mannose-containing epitope on their cell surface that is recognized by monoclonal antibody Lan3-2. Previously, the elaboration of sensory arbors in the synaptic neuropil of CNS ganglia was experimentally shown to depend on this mannose marker. Sensory arbors were abolished by perturbing sensory afferents in the intact nervous system with Lan3-2 Fab fragments, a glycosidase, or mannose-BSA. To understand the cytological mechanisms underlying mannose specific recognition for synaptogenesis, we have now studied the effects of antibody perturbation at the ultrastructural level in the sensory afferent target region. A characteristic signature of a normal sensory afferent is its profuse collateral branching, which, with ongoing development, is replaced by a single widened process, the sensory trunk, which possesses numerous synaptic vesicle clusters. The inhibition of mannose-specific recognition leads to a rapid, major reorganization of different stages of sensory afferent growth. Collateral branches at the distal growing region are reduced three- to fourfold. The pruned axons grow at an accelerated rate. Developmentally older sensory trunks experience a threefold reduction in synaptic vesicle clusters. These responses suggest that depriving sensory afferents of mannose-specific recognition aborts their synaptogenesis and causes them to resume behavior typical of tracking through axonal tracts. The current findings also suggest that the mannose marker, by promoting both collateral branching andthe proliferation of synaptic vesicle clusters, plays a critical role in two stages of sensory afferent synaptogenesis. PMID- 9740655 TI - Distinct roles of oncostatin M and leukemia inhibitory factor in the development of primordial germ cells and sertoli cells in mice. AB - Leukemia inhibitory factor (LIF) stimulates the growth of primordial germ cells (PGCs) in mouse embryo. However, as neither mice lacking LIF nor mice lacking the LIF receptor show defects in PGC growth, an alternate cytokine for PGC growth has been postulated. We investigated the role of mouse oncostatin M (mOSM), which is structurally and functionally related to LIF, in germ cell development. While LIF enhanced the survival of migratory as well as postmigratory PGCs, mOSM acted only on the postmigratory PGCs. Consistent with its biological activity, mOSM was found to be expressed in developing gonads. In the male, Sertoli cells in neonatal testis express mOSM; however, its expression is downregulated in adult testes. Moreover, mOSM enhanced the proliferation of Sertoli cells derived from neonatal testes in vitro more than human OSM or LIF. In contrast, postnatal ovaries do not express mOSM. These results indicate that mOSM is a stage- and sex specific autocrine growth factor for Sertoli cells. PMID- 9740657 TI - Loss of beta1 integrin function results in a retardation of myogenic, but an acceleration of neuronal, differentiation of embryonic stem cells in vitro. AB - Integrin cell surface receptors play an important role for cell adhesion, migration, and differentiation during embryonic development by mediating cell cell and cell-matrix interactions. Less is known about the function of integrins during commitment and lineage determination of early embryogenesis. Homozygous inactivation of the beta1 integrin gene results in embryonal death in mice around the time of implantation. In vitro, differentiation of embryonic stem (ES) cells which lack beta1 integrin (beta1-/-) into the cardiogenic lineage is delayed and results in a disordered cellular specification (Fassler et al., J. Cell Sci. 109, 2989-2999, 1996). To analyze beta1 integrin function during myogenesis and neurogenesis we studied differentiation of beta1-/- ES cells via embryoid bodies into skeletal muscle and neuronal cells in vitro. beta1-/- cells showed delayed and reduced myogenic differentiation compared to wildtype and heterozygous (beta1+/-) ES cells. RT-PCR analysis demonstrated delayed expression of skeletal muscle-specific genes in the absence of beta1 integrin. Immunofluorescence studies with antibodies against the sarcomeric proteins myosin heavy chain, titin, nebulin, and slow C-protein showed that myotubes formed, but their number was reduced and the assembly of sarcomeric structures was retarded. In contrast, neuronal cells differentiating from beta1-/- ES cells appeared earlier than wildtype and heterozygous (beta1+/-) ES cells. This was shown by the accelerated expression of neuron-specific genes and an increased number of neuronal cells in beta1-/- embryoid bodies. However, neuronal outgrowth was retarded in the absence of beta1 integrin. No functional difference between wildtype and beta1-/- cells was found with respect to secretion of gamma-aminobutyric acid, the main neurotransmitter of ES cell-derived neuronal cells. The lineage-specific effects of loss of beta1 integrin function, that is the inhibition of mesodermal and acceleration of neuroectodermal differentiation, were supported by differential expression of genes encoding lineage-specific transcription factors (Brachyury, Pax-6, Mash1) and signaling molecules (BMP-4 and Wnt-1). Because of the reduced and delayed expression of the BMP-4 encoding gene in beta1-/- cells, we analyzed in wildtype and beta1-/- cells the regulatory role of exogenously added BMP-4 on the expression of the mesodermal and neuronal marker genes, Brachyury and wnt-1, respectively. The data suggest that BMP-4 plays a regulatory role during differentiation of wildtype and beta1-/- cells by modifying mesodermal and neuronal pathways. The reduced expression of BMP-4 in beta1-/- cells may account for the accelerated neuronal differentiation in beta1-/- ES cells. PMID- 9740658 TI - Ethanol impairs migration of the prechordal plate in the zebrafish embryo. AB - Exposure of vertebrate embryos to ethanol causes cyclopia, but little is known about the underlying mechanisms of this effect. Here we show that cyclopia can be induced in the zebrafish by a short ethanol treatment during early gastrula stages and is accompanied by loss of gene expression characteristic of the ventral aspects of the fore- and midbrain. Interestingly, defects in the expression of ventral brain markers are linked to impaired migration of the prechordal plate mesoderm indicating that the correct position of the prechordal plate mesoderm under the anterior neural plate in the zebrafish embryo is required for specification of the anterior neural midline. Ethanol-induced cyclopia does not, however, impair the induction of anterior neuroectodermal structures in general. Finally, as genes like goosecoid and islet-1 are expressed in prechordal plate cells in a temporal pattern similar to control embryos despite the ectopic position of expressing cells, it appears that regulation of prechordal plate-specific gene expression is largely independent of the final position of the prechordal plate. PMID- 9740659 TI - Dredd, a novel effector of the apoptosis activators reaper, grim, and hid in Drosophila. AB - Caspases are widely conserved proteases considered to be essential effectors of apoptosis. We identified a novel Drosophila gene, dredd, which shares extensive homology to all members of the caspase gene family. Cells specified for programmed death in development exhibit a striking accumulation of dredd RNA that requires signaling by the death activators REAPER, GRIM, and HID. Furthermore, directed misexpression of each activator was sufficient to drive ectopic accumulation of dredd RNA. Heterozygosity at the dredd locus suppressed apoptosis in transgenic models of reaper- and grim-induced cell killing, demonstrating that levels of dredd product can modulate signaling triggered by these death activators. Finally, expression of REAPER, GRIM, and HID was found to trigger processing of DREDD protein precursor through a mechanism that is insensitive to, and upstream of, known caspase inhibitors. Taken together, these observations establish mechanistic connections between activators of apoptosis and a new downstream death effector in Drosophila. PMID- 9740660 TI - Expression of DjY1, a protein containing a cold shock domain and RG repeat motifs, is targeted to sites of regeneration in planarians. AB - Planarians are well-known for their exceptional regenerative abilities. This investigation focuses on the involvement of a Y-box protein, defined by the presence of a cold-shock domain, in regeneration-specific processes. Previous studies have shown that developmentally expressed Y-box proteins bind to mRNA molecules and regulate the timing of their translation. We have isolated and characterized a planarian Y-box gene, DjY1, which is specifically expressed at the site of regeneration, the blastema. DjY1 transcripts appear rapidly at the site of cutting and increase in number as the blastema grows. The timing and level of expression is similar irrespective of the orientation of the cut: in anterior, posterior, and lateral regenerative tissue. As regeneration nears completion, there is a general decrease in transcript level except in structures which are still differentiating, specifically in the auricles where new DjY1 transcripts are produced. A similarly modulated temporal pattern of expression throughout regeneration is seen in assaying the DjY1 protein. Within the population of blastemal cells, a subset of differentiating cells is specifically immunostained using antibodies to DjY1. The DjY1 protein contains a cold-shock domain and RG-repeat motifs, both of which are associated with RNA-binding properties: in vitro binding studies using recombinant DjY1 show that the preferred template is single-stranded RNA of heterogeneous sequence. These data provide the first direct evidence that a Y-box protein is involved in the regeneration process in planarians and implicate DjY1 in the translational regulation of differentiation-specific mRNAs. PMID- 9740661 TI - Disruption of primary mesenchyme cell patterning by misregulated ectodermal expression of SpMsx in sea urchin embryos. AB - The patterning of the mesoderm of the sea urchin embryo is a classical paradigm of epithelial mesenchymal interactions in organogenesis, yet little is known of its molecular basis. Here we address the role of the homeobox gene, SpMsx, a member of the highly conserved Msx gene family, in this process. Msx genes have been shown to function in the dorsoventral patterning of the central nervous system in Drosophila and in a variety epithelial-mesenchymal interactions in vertebrates. We showed previously that the SpMsx gene is expressed during embryogenesis in a complex and dynamic pattern consistent with roles in the development of subpopulations of endoderm, mesoderm, and oral ectoderm. To perturb this pattern of expression and thus probe the function of SpMsx, we injected SpMsx mRNA into single-cell zygotes and monitored development morphologically and with a series of territory-specific molecular markers. RT-PCR analysis revealed that injected SpMsx transcripts persisted at least until the gastrula stage in amounts comparable to endogenous levels. Injected embryos exhibited deficiencies in the organization of primary and secondary mesenchyme cells within the blastocoelic cavity, as well as abnormalities in spicule number and shape. Defects in the endoderm were also common, including reduced or absent archenterons. Micromere transplantation experiments revealed that the defects in skeletogenic mesenchyme patterning were non-cell autonomous, consistent with findings that cell-cell interactions between ectoderm and the progenitors of the skeletogenic mesenchyme, the primary mesenchyme cells (PMCs), are important both for PMC guidance and spicule morphogenesis. Our data, taken together with observations in other organisms on the role of Msx genes in embryonic signaling processes, particularly involving the BMP pathway, suggest that SpMsx may be a part of the mechanism by which the ectoderm influences both the arrangement of primary mesenchyme cells within the blastocoel and the shapes of the skeletal rods. PMID- 9740662 TI - Notochord regulates cardiac lineage in zebrafish embryos. AB - We focus here upon regulation by the notochord of myocardial cell fate in zebrafish. Myocardial precursors, defined by lineage tracing in the living embryo, are in the lateral plate mesoderm adjacent to the notochord-prechordal plate junction. Interestingly, the anterior end of the notochord corresponds to the posterior extent of the heart progenitor field, defined by this lineage analysis. This suggested that the notochord might suppress, or the prechordal plate might enhance, the cardiogenic fate. Nkx2.5 expression is, in the zebrafish embryo, closely correlated with the position of myocardial precursors, which reside adjacent to the notochord-prechordal plate junction. This expression, however, is extinguished in the region posterior to this junction, a region normally not contributing cells to the heart. Laser ablation of the notochord tip between the 4-somite and 12-somite stage causes posterior expansion of the Nkx2. 5-expressing region. The ntl mutation of the notochord is associated with posterior extension of Nkx2.5 expression. Lineage tracking, by laser activation of caged fluoresceinated dextran, confirms that, normally, lateral plate cells next to the notochord do not contribute progeny to the heart. After anterior notochord ablation, these cells are redirected to a heart cell fate. These data suggest that the anterior notochord delimits the posterior extent of the heart field by suppressing the heart cell fate. PMID- 9740663 TI - Conservation of the spiralian developmental program: cell lineage of the nemertean, Cerebratulus lacteus. AB - Lineage tracers were injected into individual blastomeres in embryos of the indirect-developing nemertean Cerebratulus lacteus through the formation of the fourth quartet of micromeres. Subsequent development was followed to the formation of feeding pilidium larvae to establish their ultimate fates. Results showed that these blastomeres have unique fates, and their clones give rise to highly predictable regions of the larval body. As in other spiralians, four discrete cell quadrants can be identified. For the most part, their identities are homologous to the typical spiralian A, B, C, and D cell quadrants. In some respects their fates differ from the typical spiralian fate map; however, these can be understood in terms of simple modifications of the early cleavage program. Unlike most spiralians, the first quartet micromeres in the eight-celled embryo are larger than the corresponding vegetal macromeres, and generate most of the larval ectoderm. All four of these micromeres contribute to the apical organ and generate four bilaterally situated domains of ectoderm, where the progeny of the 1a and 1d micromeres lie to the left of the median plane while those of 1b and 1c lie to the right. Unlike the progeny of the first quartet, those of the second quartet are situated in left (2a), ventral (2b), right (2c), and dorsal (2d) positions. The third quartet micromeres generate clones situated in a bilaterally symmetrical fashion similar to those of the first quartet. The alternating axial relationships exhibited by successive micromere quartets are a characteristic of spiralian development. Unlike other spiralian larvae possessing a ciliary band, the pilidium larval ciliary band is formed by all blastomeres of the first and second micromere quartets, as well as 3c and 3d. Ectomesoderm is derived from two blastomeres (3a and 3b), which give rise to the extensive array of the larval muscle cells. C. lacteus also possesses a true mesentoblast (4d) which gives rise to a pair of mesodermal bandlets, and scattered mesenchymal cells. The dual origin of the mesoderm, as both ectomesoderm and endomesoderm, appears to be a condition present in all spiralians. The gut is formed by all the fourth quartet micromeres as well as the vegetal macromeres (4A, 4B, 4C, 4D). Despite differences in the determination of axial properties and some modifications in quadrant fates, nemerteans appear to be constructed on the typical spiralian developmental platform. PMID- 9740664 TI - Sperm-egg binding in the sea urchin: a high level of intracellular ATP stabilizes sperm attachment to the egg receptor. AB - Previous studies have established that a recombinant protein fragment (45A) of the egg receptor for sperm of the sea urchin Strongylocentrotus purpuratus exhibits several characteristics that are consistent with that expected of a receptor. Using a quantitative sperm binding assay with glutathione S-transferase fused to a recombinant protein containing the C-terminal half of the 45A construct immobilized on glutathione beads, it was found that the interaction between sperm and this protein is a kinetically transient event. Sperm binding to the receptor fragment reached a maximum at 20 s after adding sperm in the presence of egg jelly to beads coated with recombinant receptor. In the next 20 120 s, approximately 50-70% of the sperm detached from the beads. Similar phenomena were observed when the kinetics of sperm binding to dejellied, glutaraldehyde-fixed eggs were studied. Because the acrosome reaction, a prelude to binding, is known to be accompanied by a decrease in the ATP level of sperm, we studied the effect of various inhibitors on both sperm detachment and the level of ATP. It was found that the detachment rate increased slightly when respiration inhibitors that blocked ATP production in mitochondria were added. In contrast, the dynein ATPase inhibitor, erythro-9-[3-hydroxynonyl]adenine, which is known to inhibit flagellum motility by blocking ATP utilization, stabilized the binding of sperm to the receptor and allowed maintenance of a high internal ATP level. Immotile, tailless sperm that physically lacked dynein ATPase, and therefore sustained their internal ATP levels, also exhibited stable binding provided that the sperm and beads were physically mixed. These results suggest that the internal ATP level of the sperm controls the stability of its binding to the receptor. The possible mechanism of the detachment and its significance with respect to the overall process of fertilization are discussed. PMID- 9740665 TI - An improved approach for construction of bacterial artificial chromosome libraries. AB - Presented here are improved methodologies that enable the generation of highly redundant bacterial artificial chromosome/P1-derived artificial chromosome libraries, with larger and relatively uniform insert sizes. Improvements in vector preparation and enhanced ligation conditions reduce the number of background nonrecombinant clones. Preelectrophoresis of immobilized high molecular-weight DNA removes inhibitors of the cloning process, while sizing DNA fragments twice within a single gel effectively eliminates small restriction fragments, thus increasing the average insert size of the clones. The size fractionated DNA fragments are recovered by electroelution rather than the more common melting of gel slices with subsequent beta-agarase treatment. Concentration of the ligation products yields a 6- to 12-fold reduction in the number of electroporations required in preparing a library of desirable size. These improved methods have been applied to prepare PAC and BAC libraries from the human, murine, rat, canine, and baboon genomes with average insert sizes ranging between 160 and 235 kb. PMID- 9740666 TI - Mapping an endometrial cancer tumor suppressor gene at 10q25 and development of a bacterial clone contig for the consensus deletion interval. AB - Frequent loss of chromosome 10q sequences in endometrial cancers suggests the involvement of a tumor suppressor gene. Previous loss-of-heterozygosity (LOH)studies have pointed to the 10q25-q26 region as the likely site of a tumor suppressor involved in endometrial tumorigenesis (S. L. Peiffer et al., 1995, Cancer Res. 55: 1922-1926; S. Nagase et al., 1996, Br. J. Cancer 74: 1979-1983; S. Nagase et al.,1997, Cancer Res. 57: 1630-1633). In an attempt to define further the localization of a tumor suppressor gene at 10q25, we screened a panel of 123 endometrioid adenocarcinomas for loss of heterozygosity of 10q25.3 sequences. Forty-three (35%) revealed LOH at one or more loci. The observed patterns of allelic loss define a minimum consensus region of deletion between D10S221 and D10S610. A sequence-ready bacterial clone contig and a long-range restriction map for a 1-Mb interval spanning the deletion region were developed as the first step in experiments directed toward the discovery the 10q25 tumor suppressor. PMID- 9740667 TI - Genomic and functional map of the chromosome 14 t(12;14) breakpoint cluster region in uterine leiomyoma. AB - A translocation involving chromosomes 12 and 14 [t(12;14)(q15;24.1)] is commonly seen in benign smooth muscle tumor as uterine leiomyoma (UL). A contig of P1 derived artificial chromosome and bacterial artificial chromosome clones on chromosome 14, encompassing a t(12;14) breakpoint cluster region (BCR) in UL, was generated principally using the recently developed HAPPY map of chromosome 14 as a framework (P. H. Dear et al., 1998, Genomics 48: 232-241). Three UL t(12;14) breakpoints have been localized within this contig, showing that a BCR of at least 400 kb exists on chromosome 14. Other studies of tumors with t(12;14) rearrangements similarly show breakpoints within a 475-kb multiple aberration region on chromosome 12. Thus t(12;14) is an example of a translocation in which the breakpoints are located within a BCR on both chromosome 12 and chromosome 14, justifying the identification of expressed sequences that are altered in these BCR regions. A total of four expressed sequences were identified in the BCR on chromosome 14. Two of these were novel cDNAs (D14S1460E and D14S1461E). The chromosome 14 cDNAs were expressed in multiple adult tissues. The identification of a large breakpoint cluster region on chromosome 14 suggests that translocations in this region mediate their effects at a distance and also that elements that predispose this region to recurrent chromosomal translocation may be widely distributed. PMID- 9740668 TI - Mutational scanning of mitochondrial DNA by two-dimensional electrophoresis. AB - An expedient, accurate, and cost-efficient test was developed to scan critical regions of the mitochondrial genome for all possible mutations by two-dimensional DNA electrophoresis. The test involves a two-step multiplex PCR amplification: a long-distance PCR to amplify almost the entire mitochondrial genome, which then serves as template for the amplification of 25 short PCR fragments in two multiplex groups corresponding to regions implicated in human diseases. The mixture of fragments was subsequently subjected to two-dimensional electrophoretic separation, first by size in a nondenaturant polyacrylamide gel and then on the basis of basepair sequence in a denaturing gradient polyacrylamide gel. This latter process of denaturing gradient gel electrophoresis is a most accurate form of mutation detection on the basis of differences in melting behavior of mutant and wildtype fragments. Evaluation of the method using samples with known homoplasmic and heteroplasmic mutations, as well as CEPH pedigrees to study segregation of polymorphic variants, indicated a very high accuracy; none of the previously identified mutations and polymorphisms escaped detection, and no erroneous segregation patterns of polymorphic variants were observed. In addition, two variants were found to be novel mutations when analyzed by sequence analysis. One of these novel mutations was a heteroplasmic mutation in the COXIII gene that was found to segregate to homoplasmy in the next generation. Heteroplasmic mutations as low as 1% of mtDNA could still be detected. PMID- 9740669 TI - Cloning and comparative mapping of the DiGeorge syndrome critical region in the mouse. AB - Chromosome deletions leading to the hemizygous loss of groups of contiguous genes are a major cause of human congenital defects. In some syndromes haploinsufficiency of a single gene causes the majority of the syndromal features, whereas other diseases are thought to be the consequences of a combined haploinsufficiency. In the case of the DiGeorge and velocardiofacial syndromes, caused by deletions within 22q11, the genetic analyses have so far failed to implicate a single gene. By virtue of FISH analysis and the creation of a BAC/P1 genomic clone contig we have mapped 19 murine homologues of genes and nine EST groups from the region deleted in DiGeorge syndrome and found them to be linked on mouse chromosome 16. Rearrangements during the divergence of mouse and human have led to differing gene orders in the two species, with implications for the most appropriate means of mimicking particular human deletions. The map confirms and extends previous analyses and the contig resources toward the generation of targeted deletions in the mouse. PMID- 9740670 TI - Blind analysis of denaturing high-performance liquid chromatography as a tool for mutation detection. AB - Denaturing high-performance liquid chromatography (DHPLC) is a novel high capacity technique for detecting new mutations. We have evaluated the sensitivity and specificity of this method in a blind analysis of exon H of the factor IX gene and exon 16 of the neurofibromatosis type 1 gene. Under a single set of conditions for each exon, 55/55 individuals carrying 48 unique mutations were correctly identified as were 55/55 individuals with wildtype alleles. We conclude that DHPLC is a highly sensitive and specific method for mutation detection. PMID- 9740671 TI - Cloning, structural characterization, and chromosomal localization of the human orthologue of Saccharomyces cerevisiae MSH5 gene. AB - We have cloned and characterized the human orthologue of the Saccharomyces cerevisiae MutS homologue 5 (MSH5) cDNA, as well as the human gene that encodes the MSH5 cDNA, as a step toward understanding the molecular genetic mechanisms involved in the biological function of this novel human protein. The identified cDNA contains a 2505-bp open reading frame (ORF) that encodes an 834-amino-acid polypeptide with a predicted molecular mass of 92.9 kDa. The amino acid sequence encoded by this cDNA includes sequence motifs that are conserved in all known MutS homologues existing in bacteria to humans. The cDNA appears, on the basis of amino acid sequence analysis, to be a member of the MutS family and shares 30% sequence identity with that of S. cerevisiae MSH5, a yeast gene that plays a critical role in facilitating crossover during meiosis. Northern blot analysis demonstrated the presence of a 2.9-kb human MSH5 mRNA species in all human tissues tested, but the highest expression was in human testis, an organ containing cells that undergo constant DNA synthesis and meiosis. The expression pattern of human MSH5 resembled that of the previously identified human MutS homologues MSH2, MSH3, and MSH6-genes that are involved in the pathogenesis of hereditary nonpolyposis colorectal cancer (HNPCC). In an effort to expedite the search for potential disease association with this new human MutS homologue, we have also determined the chromosomal location and structure of the human MSH5 locus. Sequence and structural characterization demonstrated that MSH5 spans approximately 25 kb and contains 26 exons that range in length from 36 bp for exon 8 to 254 bp for exon 25. MSH5 has been mapped to human chromosome band 6p21.3 by fluorescence in situ hybridization. Knowledge of the sequence and gene structure of MSH5 will now enable studies of the possible roles MSH5 may play in meiosis and/or DNA replicative mismatch repair. PMID- 9740672 TI - Analysis of distribution in the human, pig, and rat genomes points toward a general subtelomeric origin of minisatellite structures. AB - We have developed approaches for the cloning of minisatellites from total genomic libraries and applied these approaches to the human, rat, and pig genomes. The chromosomal distribution of minisatellites in the three genomes is strikingly different, with clustering at chromosome ends in human, a seemingly almost even distribution in rat, and an intermediate situation in pig. A closer analysis, however, reveals that interstitial sites in pig and rat often correspond to terminal cytogenetic bands in human. This observation suggests that minisatellites are created toward chromosome ends and their internalization represents secondary events resulting from rearrangements involving chromosome ends. PMID- 9740673 TI - The human Surfeit locus. AB - The organization of the human Surfeit locus containing the six sequence-unrelated housekeeping genes Surf-1 to Surf-6 (HGMW-approved symbols SURF1-SURF6) has been determined. The human surfeit locus occupies about 60 kb of DNA, and the tightly clustered gene organization and the juxtaposition of the human genes are similar to the mouse and chicken surfeit loci with the 5' end of each gene associated with a CpG-rich island. Whereas in the mouse the Surf-2 and Surf-4 genes overlap at their 3' ends, the human Surf-2 and Surf-4 genes have been found to be separated by 302 bp due to a much shorter 3' untranslated region in the human Surf-2 gene. The distance between the 3' ends of the human Surf-1 and Surf-3 genes is 374 bp, and the distance between the 5' ends of the human Surf-3 and Surf-5 genes is only 112 bp. Unusually the human Surf-5 gene contains an intron in its 5' untranslated region not found in the mouse or rat Surf-5 genes. This additional intron is also found in the Surf-5 gene of both Old and New World monkeys, being generated before the divergence of human and prosimians but after the divergence of primates and rodents. A contig of 200 kb containing the human Surfeit locus has been constructed from overlapping cosmid, P1, and PAC clones. Approximately 40 kb proximal to the 3' end of the Surf-6 gene, the 5' region of the ABO glycosyltransferase gene has been detected. This allows us to determine the orientation of the Surfeit and ABO loci with respect to each other and to the telomere and centromere of human chromosome 9. PMID- 9740674 TI - The mouse tumor necrosis factor receptor 2 gene: genomic structure and characterization of the two transcripts. AB - The mouse TNFR2 gene has been cloned, sequenced, and characterized as a gene spanning >44 kb of the genome. By alignment of five genomic clones we have established that TNFR2 consists of 10 exons and 9 introns with exons ranging in size from 35 bp to 2.6 kb and introns ranging from 322 bp to >16 kb. All splice acceptor and donor sites conform to the canonical AG/GT rule. The translation initiation and termination sites are located in exon 1 and 10, respectively. Although TNFR2 lacks a canonical TATA box, the gene is transcribed from a unique start site located 70 bp upstream of the ATG initiation codon that conforms to the consensus Inr motif. Several cis-elements for transcription factors were identified in the 5' flanking region, including NF-1, Sp-1, AP2, gamma-IRE, and NF-kappaBeta motifs. Functional analysis indicates that the region -705/-412 contains a negative cis-acting element and that the minimal promoter contains motifs that confer LPS inducibility. Two mouse TNFR2 mRNAs of 3.2 and 4.1 kb are detected by Northern blot analysis, but until now their origin has not been explained. No evidence of alternative splicing of the coding exons was found. However, hybridization studies and amplification of cDNA ends suggest the use of a noncanonical polyadenylation signal in the untranslated region of exon 10. A comparative analysis of the 3' untranslated regions of the human and mouse TNFR2 genes shows highly divergent 3' ends. The possibility of an ancestral mouse TNFR2 mRNA similar to the short transcript is discussed. PMID- 9740675 TI - Isolation and characterization of human SGT and identification of homologues in Saccharomyces cerevisiae and Caenorhabditis elegans. AB - We have recently isolated a rat cDNA encoding a novel cellular protein able to interact with the major nonstructural protein NS1 of parvovirus H-1 and have termed this protein SGT, for small glutamine-rich tetratricopeptide repeat (TPR) containing protein. Here we report the isolation of a cDNA from human placenta encoding the human homologue, human SGT. SGT from rat and human contain 314 and 313 amino acids, respectively, and share 91% sequence identity at the protein level. The highest degree of similarity is present within the central region containing three TPR motifs in tandem array. The similarities, however, also extend beyond this region. Human SGTtranscript was found to be ubiquitously present in all human tissues tested. By fluorescence in situ hybridization analysis we have mapped the human gene to chromosome 19p13. The SGT-coding sequences are evolutionarily conserved, since we could identify genes encoding proteins of similar size and structure in the genomes of Saccharomyces cerevisiae and Caenorhabditis elegans. PMID- 9740676 TI - Mapping of a novel human carbonyl reductase, CBR3, and ribosomal pseudogenes to human chromosome 21q22.2. AB - To find the genes contributing to Down syndrome, we constructed a 4-Mb sequence ready map spanning chromosome 21q22.2 with megabase-sized cosmid/P1-derived artificial chromosome (PAC) contigs. The restriction map with rare cutting enzymes, followed by sequencing from the clustering sites, has defined CpG islands and revealed the genes associated with CpG islands (Accession No. D85771). Of these, two human carbonyl reductases (CBR; EC1.1.1.184) were found in a PAC 25P16 clone. CBR catalyzes the reduction of a large number of biologically and pharmacologically active carbonyl compounds to their corresponding alcohols and has been mapped in 21q22.1. To confirm these results, we sequenced the PAC clone in shotgun strategies and identified a novel carbonyl reductase, designated CBR3, 62 kb downstream from the original CBR. In addition, three ribosomal pseudogenes, L23a, S9, and L3, and some cDNAs with ESTs were mapped in the sequence. In conclusion, the sequence analysis for CpG islands predicted from the megabase-sized contigs will reveal and identify the genes involved in Down syndrome. PMID- 9740679 TI - Keratocan (Kera), a corneal keratan sulfate proteoglycan, maps to the distal end of mouse chromosome 10 PMID- 9740680 TI - Localization of p27 beta4 binding protein gene (ITGB4BP) to human chromosome region 20q11.2 PMID- 9740681 TI - Developmental genomics and its relation to aging PMID- 9740677 TI - Isolation and characterization of two novel metalloproteinase genes linked to the Cdc2L locus on human chromosome 1p36.3. AB - The terminal end of the short arm of human chromosome 1, 1p36.3, is frequently deleted in a number of tumors and is believed to be the location of multiple tumor suppressor genes. Thus far, a bona fide tumor suppressor gene from this region has not been identified. The isolation and characterization of new 1p36 genes is, therefore, of some interest. Two novel matrix metalloproteinase genes, MMP21 and MMP22, have been identified in the Cdc2L1-2 locus, which spans approximately 120 kb on 1p36.3. These genes encode novel metalloproteinases that contain prepro, catalytic, cysteine-rich, interleukin-1 receptor-related, and proline-rich domains. Their catalytic domains are most closely related to stromelysin-3 and contain the consensus HEXXH zinc-binding region required for enzyme activation, while their cysteine-rich domains appear to be related to a number of human, mouse, and Caenorhabditis elegans metalloproteinase sequences. Of some possible interest is the absence of a highly conserved cysteine residue in the proenzyme domain, the so-called "cysteine switch," which has been shown to be involved in the autocatalytic activation of many metalloproteinases. The MMP genes are located less than 1 kb from the 3' regions of Cdc2L1 and Cdc2L2, suggesting that the MMP and Cdc2L genes are part of a larger region that has been duplicated. Finally, the MMP21/22 genes express multiple mRNAs, some of which are derived by alternative splicing, in a tissue-specific manner. PMID- 9740682 TI - SGO: a continually evolving society. PMID- 9740683 TI - Staging and therapeutic value of lymphadenectomy in endometrial cancer. PMID- 9740685 TI - Radicality in gynecologic cancer surgery: a historical perspective. PMID- 9740684 TI - Long-term outcomes of therapeutic pelvic lymphadenectomy for stage I endometrial adenocarcinoma. AB - OBJECTIVE: The treatment of patients with stage I endometrial adenocarcinoma is often shorter and less expensive if total abdominal hysterectomy (TAH), bilateral salpingo-oophorectomy (BSO), and therapeutic lymphadenectomy are used rather than TAH, BSO, pelvic lymph node sampling, and pelvic external beam radiation. We studied whether the survival and morbidity of patients treated with therapeutic lymphadenectomy are equal to or better than with these alternative treatments. METHODS: We reviewed the medical records of patients with stage I endometrial adenocarcinoma who were enrolled in the MetroHealth Medical Center tumor registry between 1970 and 1993 after undergoing full pelvic lymph node dissection, in addition to total abdominal hysterectomy, bilateral salpingo-oophorectomy, and vaginal brachytherapy. The mean number of resected nodes was 33 (median, 31; interquartile range, 19). Patients were followed for 1. 6-20 years (median, 8 years; interquartile range, 5.8 years). Morbidity and survival rates were compared to published series using similar treatment strategies and to those from studies using pelvic external beam radiation and pelvic lymph node sampling rather than lymphadenectomy. RESULTS: Of 192 patients with pathologic stage I (FIGO 1988) endometrial adenocarcinoma, 178 patients had full pelvic lymph node dissection; 159 patients were evaluable. The 15-year overall survival was 98%; 10 and 15- year disease-free survivals were 96 and 94%, respectively. Overall morbidity was 18% (29/159), and moderate-to-severe morbidity was 13% (21/159). Recurrences were seen in 4.4% (7/159) of patients. Grade and myometrial invasion were not significant predictors of disease-free survival after full pelvic lymph node dissection (grade, P = 0.42; stage, P = 0.67). The results compare favorably with those of similar studies and with studies of pelvic external beam radiation. CONCLUSIONS: Primary surgical management with total abdominal hysterectomy, bilateral salpingo-oophorectomy, therapeutic pelvic lymphadenectomy, and vaginal brachytherapy is a viable and possibly preferable option for patients with stage I endometrial adenocarcinoma. PMID- 9740687 TI - Ovarian metastasis in endometrial carcinoma. AB - A retrospective study was conducted to investigate the clinical significance of ovarian metastasis in 439 patients with clinical stage I endometrial cancer surgically treated by performing total abdominal hysterectomy, bilateral salpingo oophorectomy, and pelvic lymphadenectomy. Histologic examination revealed that 22 patients (5%) had ovarian metastasis. The maximum diameter of the ovarian metastases ranged from 1 to 100 mm. In 18.2% (4/22) of patients with ovarian metastasis, the maximum diameter was less than 2 mm. Patients with metastasis limited to the ovarian surface showed 100% positive peritoneal cytology, 0% lymph node metastases, and 50% recurrence, while patients with metastasis inside the ovary showed 10% positive peritoneal cytology, 36% lymph node metastases, and 53% recurrence. The prognosis of patients with ovarian metastasis alone was situated midway between that of patients with cancer limited to the uterus and that of patients with lymph node metastasis alone. The lymph node status was of importance to determine the prognosis of patients with ovarian metastasis. The series also suggests that there may be two routes for ovarian metastasis; one is a route via the fallopian tube to the ovarian surface and the other is a route via the lymphatics to the inside of the ovary. PMID- 9740686 TI - Multiple drug resistance parameter expression in ovarian cancer. AB - OBJECTIVE: Drug resistance represents a complex problem for the treatment of ovarian cancer. This study was undertaken to assess several putative resistance parameters (DRP) in parallel in cancer tissue from newly diagnosed patients with ovarian cancer in order to establish possible correlations to known clinical factors and prognosis. MATERIAL AND METHODS: Tumor and adjacent tumor free ovarian tissue samples from 39 consecutive, untreated female patients with ovarian cancer were obtained and as potential DRPs, the level of glutathione (GSH), the activities of glutathione S-transferase (GST), glutathione-peroxidase (GPx), O6-alkylguanine-DNA alkyltransferase (Atase), and topoisomerase II (TOPO) were assessed biochemically, and P-glycoprotein (Pgp) was assessed by Western blotting. RESULTS: Interindividual variations were high and each patient exhibited an individual profile of resistance factor expression. Levels of GSH were increased with stage (linear trend: P < 0.002), and GST, GPx, and Atase showed a similar tendency. With few exceptions no correlation was found between the DRPs and other prognostic characteristics. All tested DRPs except Pgp showed significantly higher levels/activities in tumor tissues than in the surrounding tumor free tissues (P < 0.05). The tested DRPs were found not to influence response to treatment. CONCLUSIONS: It is concluded that elevated DRPs reflect an intrinsic pattern of components of the detoxifying system in the tumor tissue. This pattern differs between patients and may partly explain the difficulty to assess the clinical importance of individual DRPs in order to translate them into recommendations for specific therapies. PMID- 9740688 TI - Steroid receptor expression in endometria from women treated with tamoxifen. AB - Breast cancer patients receiving tamoxifen (Tam) are at an increased risk for developing endometrial carcinomas, possibly due to the partial estrogenic effect of Tam on endometrial cells. Progestational therapy has not routinely been included in Tam regimens. It was our aim to determine the presence of estrogen receptors (ERs) and progesterone receptors (PRs) in normal and abnormal endometria from postmenopausal women with breast cancer who were treated with Tam. Standard immunohistochemical staining of ERs and PRs was performed on paraffin sections from formalin-fixed uterine curettings or hysterectomy specimens from 40 patients who had received 20-40 mg of Tam daily for a minimum of 3 months. For comparison, normal endometria from 20 women who had not received Tam (11 premenopausal, 9 postmenopausal) were also studied for ER and PR expression. Staining was evaluated using semiquantitative immunoreactivity scores (IRS) ranging from 0 (negative) to 12 (strongly positive). In the group of patients receiving Tam, ERs and PRs were detected in the nuclei of glandular cells in 24/24 cases of endometrial atrophy (ER/PR-IRS, 2-12), in 8/8 endometrial polyps (ER-IRS, 6-12; PR-IRS, 4-12), in 4/4 adenomatous endometrial hyperplasias (ER-IRS, 3-8; PR-IRS, 1-12), and in 4/4 well-differentiated endometrioid adenocarcinomas (ER-IRS, 2-12; PR-IRS, 6-8). Of the 11 endometria from premenopausal patients who had not received Tam, 8 were ER+/PR+ (ER-IRS, 1-12; PR IRS, 1-12), 1 was ER+/PR- (ER-IRS, 3; PR-IRS, 0), 1 was ER-/PR+ (ER-IRS, 0; PR IRS, 2), and 1 was ER-/PR- (ER/PR-IRS, 0). Among 9 atrophic endometria from women not treated with Tam, 6 were ER+/PR+ (ER-IRS, 4-12; PR-IRS, 3-6), 1 was ER+/PR- (ER-IRS, 4; PR-IRS, 0), and 2 were ER-/PR- (ER/PR-IRS, 0). The consistent finding of ER and PR expression in endometria from postmenopausal women receiving Tam further supports the suspected estrogenic effect exerted by Tam on endometrial cells. Progestational therapy could be beneficial in the prevention of Tam induced abnormal endometrial proliferations. PMID- 9740689 TI - Clinical outcome of micrometastasis in the lung in stage IA persistent gestational trophoblastic disease. AB - BACKGROUND: Computed tomography (CT) of the thorax can be used in the staging of persistent gestational trophoblastic disease (PGTD). However, the prognostic significance of micrometastasis in the lung detected by CT of the thorax has not been well documented. The aim of the study is to define the effect of micrometastasis on the clinical course of the disease. METHODS: Thirty-five patients who had nonmetastatic GTD underwent CT thorax examination before treatment in the Department of Obstetrics and Gynaecology, University of Hong Kong. All patients had workups which showed no evidence of metastasis and were diagnosed as FIGO stage IA. They all received methotrexate (MTX) infusion therapy. RESULTS: Three groups of patients were identified based on the thorax CT findings. Sixteen patients (45.7%) showed no evidence of micrometastasis on CT thorax. Two of them (12.5%) had poor response to MTX with unsatisfactory fall in serum hCG levels requiring change of chemotherapy to actinomycin D. Nine patients had suspicious micrometastasis and one (11.1%) of them needed change of MTX. Ten patients had micrometastasis and one (10%) of them needed change of MTX. There was only one recurrence and it was in the suspicious micrometastasis group (11.1%). There was no statistically significant difference in the rate of poor drug response or recurrence among the three groups of patients. CONCLUSIONS: Micrometastases in the lung do not affect the clinical outcome of patients with FIGO stage IA disease. CT thorax is not essential in the staging of GTD. PMID- 9740690 TI - Effects of retinoic acid combined with irradiation on the expression of major histocompatibility complex molecules and adhesion/costimulation molecules ICAM-1 in human cervical cancer. AB - OBJECTIVES: Radiation treatment is one of the most standardized and effective modalities for contemporary cervical cancer therapy. In addition, the radiation potentiating effects of retinoic acid have been recently described. In order to investigate whether enhanced immunogenicity might be responsible for such potentiation, we have evaluated the effects of retinoic acid combined with high doses of gamma-irradiation on the expression of major histocompatibility complex (MHC) Class I and II and intercellular adhesion molecule-1 (ICAM-1) in human cervical carcinoma cell lines. METHODS: The expression of surface antigens (MHC Class I and II and ICAM-1) was evaluated by FACS analysis in untreated control cells and in cells following their exposure to retinoic acid, high doses of gamma irradiation (i.e., 5000 and 10,000 cGy), or the combination of the two procedures. RESULTS: HT-3 and SiHa cervical cancer cells expressed variable levels of MHC Class I and ICAM-1 antigens while Class II surface antigens were not detectable. Exposure to either 5000 or 10,000 cGy completely inhibited cell replication in both cell lines and significantly and consistently increased the expression of all surface antigens present on the cells prior to irradiation. Irradiation was unable to induce neoexpression of antigens previously not expressed by these cells (i.e., MHC Class II). In a similar fashion, retinoic acid was also able to significantly increase the expression of MHC Class I and ICAM-1 antigens when compared to untreated tumor cells but was not able to induce the expression of HLA Class II surface antigens. Exposure to the combination of radiation plus retinoic acid significantly upregulated HLA Class I and ICAM-1 molecules in an additive manner when compared to the levels obtainable with the exposure to radiation or retinoic acid alone. CONCLUSIONS: These data indicate that the combination of these two treatments could induce an additive effect on the expression of immunologically important surface antigens in human cervical cancer cells. These findings, together with the powerful antiproliferative effect of retinoids and irradiation on tumor cells, suggest that the combined regimen may be a promising and more effective combination for the treatment of cervical cancer. PMID- 9740691 TI - Transforming growth factor-alpha and insulin-like growth factor-I, but not epidermal growth factor, elicit autocrine stimulation of mitogenesis in endometrial cancer cell lines. AB - OBJECTIVES: Endometrial carcinoma cell lines were evaluated for epidermal growth factor (EGF), transforming growth factor-alpha (TGF-alpha), and insulin-like growth factor I (IGF-1) production and for autocrine stimulation. METHODS: Conditioned, serum-free media (CM) from cell lines RL95-2, KLE, HEC, and Ishikawa (ISH) were concentrated radioimmunoassayed (RIA). Samples for the IGF-1 assay were extracted with acid-ethanol to remove IGF-1 binding protein. Polymerase chain reaction (PCR) was used to validate the presence of mRNA for growth factors and receptors. Cells were incubated with Ab528, an antibody blocking EGF receptors, and alphaIR3, an antibody blocking IGF-1 receptors. Proliferation was quantified using [3H]thymidine incorporation. RESULTS: TGF-alpha was detected in CM: RL95-2 (0.4 +/- 0.001 ng/ml), KLE (0.7 +/- 0.003 ng/ml), HEC (0.8 +/- 0.01 ng/ml), ISH (1.2 +/- 0.05 ng/ml). No EGF was detected in CM. In extracted samples, IGF-1 was detected in CM: RL95-2 (0.8 +/- 0. 03 ng/ml), KLE (1.25 +/- 0.02 ng/ml), HEC (1.6 +/- 0.01 ng/ml), ISH (1.6 +/- 0.08 ng/ml). Unconditioned media served as the control. EGF, TGF-alpha, and IGF-1 mRNA was identified in all cell lines, as was the mRNA for EGF and IGF-1 receptors. Incubation with Ab528 or alphaIR3 resulted in significant inhibition of DNA synthesis in HEC 1A, KLE, and ISH. No inhibition was detected in the RL95-2 cell line. A control antibody did not inhibit the cell lines. CONCLUSION: Autocrine production and stimulation of endometrial carcinoma cell lines by TGF-alpha and IGF-1 are demonstrated in three of four endometrial cancer cell lines. No measurable EGF was produced by any of the cell lines. PMID- 9740692 TI - Thrombocytosis as a prognostic factor in endometrial carcinoma. AB - We reviewed the prevalence of thrombocytosis (platelet count >/=400, 000/microL) and its association with outcome in 135 consecutive endometrial carcinoma patients and compared the platelet count with other prognostic factors. Nineteen of 135 patients (14%) had thrombocytosis. Thrombocytosis was significantly more frequent in advanced disease (stage II-IV), unfavorable grade (G2 and G3), deep myometrial invasion, and lymph-vascular space invasion. The overall 5-year survival rate was 92%. The 5-year survival rate of the 19 patients with thrombocytosis was significantly worse than that of the patients without thrombocytosis (61 vs 96%, P < 0.0001). The recurrence rate was significantly higher in patients with thrombocytosis than in those with a platelet count <400,000/microL (7 vs 32%, P < 0.005). In a multivariate analysis, thrombocytosis continued to be a predictor of worse prognosis. In conclusion, we found thrombocytosis to be a prognostic factor for survival in patients with endometrial carcinoma. PMID- 9740693 TI - A phase II study of prolonged oral etoposide in advanced or recurrent carcinoma of the cervix. AB - OBJECTIVE: To evaluate the efficacy and toxicity of prolonged oral etoposide as single agent chemotherapy in patients with advanced or recurrent carcinoma of the cervix. METHODS: Between May 1991 and February 1993, 44 patients with advanced or recurrent carcinoma of the cervix were entered onto this study. Patients were eligible if they had received no more than two prior cytotoxic regimens. The initial dose of etoposide was 37.5 mg/m2 administered orally on a daily basis on days 1-21 of a 28-day cycle. Subsequent doses were unchanged, reduced, escalated, or omitted according to toxicity. Patients were evaluated for response and toxicity using standard Gynecologic Oncology Group criteria. RESULTS: Forty-four patients were evaluable for response and toxicity. The overall response rate was 9.1% (2 CR, 2 PR). In patients with no prior chemotherapy the response rate was 4/25 compared to 0/19 for those who had prior therapy. The mean response duration was 2.7 months and the median survival from treatment for all patients was 7.7 months. The major toxicity was granulocytopenia, with 11% of patients having grade 3 or 4 toxicity. Gastrointestinal toxicity of some degree occurred in 11% of patients, and alopecia was universal. CONCLUSION: Prolonged oral etoposide has limited activity in advanced or recurrent carcinoma of the cervix. Its use as palliative therapy for this disease is not indicated. PMID- 9740694 TI - A clinicocytopathologic study of adenoma malignum of the uterine cervix. AB - OBJECTIVES: To investigate the nature and the clinical course of adenoma malignum (the so-called minimal deviation adenocarcinoma) of the uterine cervix by conducting a retrospective study of 6 cases consecutively treated in a single institute. METHODS: The pathologic classification of adenoma malignum was performed according to the WHO classification (1994). RESULTS: These tumors accounted for only 1.32% (6/453) of invasive cervical adenocarcinomas. All the cases showed either a watery discharge or atypical genital bleeding, or both, at the time of diagnosis. The preoperative cytologic diagnosis of adenoma malignum was made in 83.3% (5/6) of cases. The preoperative punch biopsy, on the other hand, failed to confirm the diagnosis of adenoma malignum in all cases, although the presence of the disease was suspected in 2 cases (33%). The 5-year survival rate and 5-year disease-free survival rate were 100 and 83.3%, respectively. CONCLUSIONS: This series demonstrates that cytologic examination is a potent screening method to detect this rare disease. When the presence of this disease is suspected by the cytologic examination, a deep biopsy is necessary to make an accurate diagnosis. An ordinary cervical biopsy is usually insufficient to detect deeply positioned tumor glands. The prognosis of the disease may be better than that for ordinary cervical adenocarcinoma. PMID- 9740695 TI - Factors predictive of survival after first relapse or progression in advanced epithelial ovarian carcinoma: a prediction tree analysis-derived model with test and validation groups. AB - OBJECTIVE: To identify factors predictive of overall survival after first relapse or primary progression in patients with advanced epithelial ovarian cancer. METHODS: "Tree-structured prediction of survival for censored survival data" was used to identify the independent prognostic factors in the test group (n = 352) who were the patients from the previously reported Canadian OV.8 trial. A prognostic model was developed using these factors and subjected to validation in the Canadian OV.4 trial cohort (n = 282). RESULTS: Based upon three factors, time from diagnosis to first recurrence or progression, tumor grade at diagnosis, and ECOG performance status at original diagnosis, three groups of patients were identified. These were labeled as good, intermediate, and poor prognosis with median survivals post relapse of 18 (12), 6 (5), and 1 (2) months, respectively. The figure in parentheses is the survival in the validation cohort. CONCLUSIONS: These prognostic groupings enable us to recommend second-line treatment more logically. The patients in the poor prognosis group have such a limited survival that cancer shrinking therapy should not routinely be offered. In addition the use of the individual predictive factors as stratification factors will help to avoid erroneous conclusions about treatment efficacy. PMID- 9740696 TI - Relationship between peritoneal washing cytology through implantable port system (IPS-cytology) and second-look laparotomy in ovarian cancer patients with unmeasurable residual diseases. AB - OBJECTIVE: Intraperitoneal chemotherapy for minimal residual ovarian cancer has been shown to be effective. However, evaluation of the response without a second look laparotomy (SLL) is impossible because such disease is unmeasurable by radiographic studies. In this prospective pilot study, we evaluated the relationship between implantable port system (IPS)-cytology and findings by SLL in patients with unmeasurable diseases. METHODS: Patients eligible for this study were those who had either unmeasurable residual disease at the time of initial surgery or measurable residual disease which became unmeasurable before second look laparotomy. At the time of the initial surgery, the IPS was placed and intraperitoneal chemotherapy was administered. To obtain IPS-cytology, approximately 500 ml of saline was infused, with the patients changing their position to wash the peritoneal surface as thoroughly as possible. More than 20 ml of saline was recovered for cytological evaluation. SLL was performed after 4 to 10 courses of chemotherapy. Results of IPS-cytology obtained immediately before SLL and the SLL findings were compared. RESULTS: Forty-four patients were entered into this study. Twenty-nine patients had unmeasurable residual disease at the time of initial surgery and 15 had measurable residual disease which became unmeasurable before SLL. Only 40 patients were eligible for evaluation because catheter failure occurred in 4 patients. Three of the 26 patients who had negative IPS-cytology results were found to have positive SLL results. All 14 patients who had positive IPS-cytology results also had positive SLL results. CONCLUSIONS: IPS-cytology can detect intraperitoneal persistent disease in patients with unmeasurable residual ovarian cancer. Persistent positive IPS cytology can indicate a negative response to chemotherapy, thus making it possible to avoid SLL. Further study with a larger number of patients is required to determine the role of negative IPS-cytology. PMID- 9740697 TI - Neoadjuvant chemotherapy with mitomycin C, etoposide, and cisplatin for adenocarcinoma of the cervix. AB - Between May 1990 and February 1995, 16 patients with adenocarcinoma or adenosquamous carcinoma of the uterine cervix were prescribed neoadjuvant chemotherapy consisting of cisplatin (50 mg/m2) on day 1, mitomycin C (10 mg/m2) on day 1, and etoposide (100 mg/m2) on days 1, 3, and 5 (MEP). In 2 patients stage was IB1, 5 were in stage IB2, 1 was in stage IIA, 5 were in stage IIB, 2 were in stage IIIB, and one was in stage IVB. A median of three courses of chemotherapy was given (range two to five). Of the 16 patients, 3 had a complete response and 5 had a partial response (response rate, 50%). Following termination of this chemotherapy, 12 patients with stage I or stage II carcinoma underwent radical hysterectomy. Three were given adjuvant radiotherapy because of positive pelvic nodes. One stage IIB patient, 1 stage IIIB patient and 1 stage IVB patient underwent standard radiotherapy and 1 stage IIIB patient underwent chemotherapy with another regimen because MEP therapy was without effect. Histopathological examinations revealed that changes as a result of the chemotherapy correlated well with clinical responses. Moderate or marked pathological changes occurred in 3 with a clinically complete response. The mean survival period of responders was 47.5 months while that of nonresponders was 28.3 months. Side effects of chemotherapy with MEP were within acceptable limits. The dose-limiting toxicity was myelosuppression and for only 1 patient the dose was reduced because of thrombocytopenia. Our preliminary study indicates that this chemotherapy regimen is effective for subjects with adenocarcinoma of the cervix. A prospective cooperative group trial on this regimen is ongoing. PMID- 9740698 TI - Radiation therapy of pelvic recurrence after radical hysterectomy for cervical carcinoma. AB - OBJECTIVES: To evaluate the efficacy of radiation therapy and potential prognostic factors in patients treated for pelvic recurrence of cervical carcinoma after radical hysterectomy. MATERIALS: The records of 50 patients treated between 1964 and 1994 for an isolated pelvic recurrence of cervical carcinoma a median of 10.5 months after initial radical hysterectomy were retrospectively reviewed. Patients were categorized according to the extent of disease on clinical examination as group 1, mucosal involvement only (5); group 2, paravaginal extension (11); group 3, central recurrence with pelvic wall extension (13); and group 4, recurrences limited to the pelvic sidewall (21). Seven patients with group 3 or 4 disease who had a poor performance status were treated with palliative intent using hypofractionated radiotherapy. The remaining 43 patients were treated with curative intent, 33 with radiotherapy only and 10 with a combination of cisplatin-based chemotherapy and radiotherapy. Survival rates were calculated from the date of initial recurrence. Median follow-up of surviving patients was 109 months. RESULTS: The overall 5-year survival rate was 33% for all 50 patients (median survival, 18 months), 39% for the 43 patients treated with curative intent, and 25% for patients with isolated sidewall recurrences treated with curative intent. The survival rate was 69% for patients with group 1 and 2 disease and 18% for those treated with curative intent for group 3 disease (P = 0.07). The survival rate was better for patients with recurrent squamous carcinomas (51%) than for those with adenocarcinomas (14%) (P = 0. 05). Three group 4 patients who survived more than 5 years were treated with external-beam radiation alone. Eight-one percent of patients who had a second recurrence had evidence of disease progression. Three patients experienced late treatment complications. CONCLUSIONS: Patients who experience an isolated recurrence of cervical cancer after initial radical hysterectomy have an excellent prognosis if disease does not involve the pelvic wall. Occasional long term survivors of recurrent disease involving the pelvic wall justify an aggressive treatment approach. PMID- 9740699 TI - The role of adjuvant radiotherapy in carcinoma of the endometrium-results in 550 patients with pathologic stage I disease. AB - OBJECTIVES: A retrospective analysis of 550 women with pathological stage I carcinoma of the endometrium who were seen between January 1984 and December 1988 was performed in order to assess the value of adjuvant radiation therapy. METHODS: Two-hundred twenty-eight patients were treated with surgery alone (S); 97 received adjuvant external beam radiotherapy (S + EXT); 217 received external beam radiotherapy and colpostats (S + EXT + IC); and 8 patients received only colpostats (S + IC). Pelvic radiation therapy, usually 40 Gy in 20 fractions, was administered to 94% of patients whose tumors showed greater than 50% myometrial invasion and to 89% of patients with FIGO grade 3 tumors. Colpostats were used in 40% of patients, the majority of whom had lower uterine segment involvement. RESULTS: The overall survival rate for the whole group using Kaplan-Meier estimates was 84% at 5 years. The 5-year overall survival rates for each treatment group, excluding the S + IC group, were 90% for S alone, 79% for S + EXT, and 82% for S + EXT + IC. The 5-year disease-free survival rates were 84, 77, and 77%, respectively. Local control rates at 5 years were 93, 94, and 95% in the three treatment groups, but the patterns of relapse were different. Distant metastases occurred more frequently among the patients who received adjuvant radiation therapy (36/49, 73%) than among those who did not (4/19, 21%). Late toxicity was documented in 66 patients. Twelve patients had EORTC/RTOG grade 3 and 4 complications; all had been treated with S + EXT + IC. FIGO grade (P = 0.009), lower uterine segment involvement (P = 0.009), and age (P = 0.03) were significant predictors of worse disease-free survival in a multiple regression analysis. CONCLUSIONS: The addition of vaginal vault brachytherapy to external beam radiotherapy did not appear to improve local cure rates nor survival, but increased the incidence of late radiation toxicity. PMID- 9740700 TI - Clinical characteristics of clear cell carcinoma of the ovary. AB - OBJECTIVE: The aim of this study is to evaluate the clinical characteristics of clear cell carcinoma of the ovary. METHODS: Between 1986 and 1996, 45 patients with clear cell carcinoma of the ovary were identified by scanning the medical records department and the tumor registry at our institution. RESULTS: Median age was 55 years (range 31-80 years). Tumors were 60% (27/45) stage I, 11% (5/45) stage II, 20% (9/45) stage III, and 9% (4/45) stage IV. All patients presented with a pelvic mass ranging in size from 2 x 3 to 20 x 30 cm and all except 1 had optimal cytoreduction. All patients received postoperative platinum-based chemotherapy, 47% (21/45) in combination with paclitaxel. One stage Ia patient refused therapy. Of the 6 stage III/IV patients with measurable residual tumor, 67% (4/6) partially responded to first line chemotherapy by CT scan or second look laparotomy. Recurrences occurred in 37% (10/27) stage I patients, including 18% (2/11) stage Ia, 33% (1/3) stage Ib, and 54% (7/13) stage Ic. Time to recurrence was 16 and 38 months for the two stage Ia patients and 35 months (median, range 18-56 months) for the stage Ic patients. Survival after recurrence was significantly related to disease-free interval after primary chemotherapy. With a median follow-up of 40 months (range 4-145 months), 93% (25/27) of stage I patients are alive, 20% (5/25) with disease, while 46% (6/13) of stage III/IV patients are alive. Median survival for the stage III/IV patients was 22 months (range 4-70 months). CONCLUSIONS: Clear cell tumors of ovary frequently present at early stages. However, these tumors have a propensity for recurrence even after primary chemotherapy in early stage tumors. PMID- 9740701 TI - Risk of venous access device wound complications in patients undergoing paclitaxel chemotherapy for gynecologic malignancies. AB - OBJECTIVE: To determine if wound complications after placement of a central venous catheter access device are related to the type of postsurgical cytotoxic chemotherapy administered. METHODS: All patients in a 10-year period undergoing placement of central venous access device followed by postsurgical chemotherapy for gynecologic malignancies were included in this retrospective case-control study. RESULTS: Sixty-eight patients underwent 78 placement procedures followed by chemotherapy. Six catheters (7.7%) in five patients developed wound complications. Variables evaluated included the type of gynecologic malignancy, previous use of chemotherapy, patient age and weight, preoperative white blood cell count, type of access device and insertion site, use of prophylactic antibiotics, type of chemotherapy and interval to administration, development of wound complication, and catheter removal. Univariate analysis shows an association between subsequent catheter site wound complication and paclitaxel use (P = 0.02) as well as wound complication and combined paclitaxel and cisplatin use (P = 0.005). Multivariate analysis with stepwise linear regression confirms that a paclitaxel containing regimen is associated with an increase in wound breakdown (P = 0.04). CONCLUSION: The use of a paclitaxel containing chemotherapeutic regimen administered after placement of an indwelling central venous access device in gynecologic oncology patients is associated with wound complications of the catheter site. PMID- 9740702 TI - Prolonged oral etoposide in recurrent or advanced squamous cell carcinoma of the cervix: a gynecologic oncology group study. AB - OBJECTIVE: Previous studies by the Gynecologic Oncology Group have demonstrated no activity with bolus etoposide in squamous cell carcinoma of the cervix. Prolonged oral etoposide, which exploits the schedule dependency of this agent, has demonstrated activity in non-small cell carcinoma of the lung and has been studied in combination therapy with cisplatin. To evaluate prolonged oral etoposide in previously treated squamous cell carcinoma of the cervix, the current Phase II trial was conducted. METHODS: Eligibility included squamous cell cancer of the cervix, measurable disease, allowed no more than one prior chemotherapy regimen which did not include etoposide, WBC >/=3000/microliter, platelet count >/=100, 000/microliter, serum creatinine /= 3000/microliter, platelet count >/=100, 000/microliter, serum creatinine 15 years) with a positive HIV serology (confirmed by Western blot) that had been revealed by a skin disease seen at the Marchoux Institute at Bamako between June 1991 and September 1994 were included in the study. RESULTS: Two hundred sixty-three skin diseases revealed 233 cases of HIV infection. Diseases observed were: zoster (n = 71), seborrheic dermatitis (n = 43), Kaposi's sarcoma (n = 34), prurigo (n = 31), sexually transmitted diseases (n = 27), extensive dermatophytosis (n = 12), psoriasis (n = 12), molluscum contagiosum (n = 8), acquired ichthyosis (n = 3), cutaneous leishmaniasis (n = 2) and other skin diseases (n = 10). More than one disease were associated in 28 patients. Certain particular features were noted (superinfection of zoster, papular margin in dermatophytosis). DISCUSSION: In Africa, certain skin diseases often reveal HIV infection and some diseases have a high positive predictive value for HIV infection (zoster, seborrheic dermatitis, prurigo, Kaposi's sarcoma, extensive dermatophytotis). For prognosis, frequently associated diseases are signs of AIDS (Kaposi's disease, prurigo, molluscum contagiosum). PMID- 9740825 TI - [Treatment of discoid lupus erythematosus with sulfasalazine: 11 cases]. AB - INTRODUCTION: Antimalaria agents and thalidomide are two reference drugs for discoid lupus erythematosus. In non-responders or after secondary resistance or contraindications, there are a number of alternative therapeutics which are less effective and more toxic. We therefore conducted an open study in patients with discoid lupus erythematosus treated with sulfasalazine. PATIENTS AND METHODS: Seven men and four women (mean age 40 years) with severe discoid lupus erythematosus (mean duration of disease 14 years) were treated with sulfasalazine (2 g/d). This treatment was initiated after a previous failure or contraindication of antimalarial drugs or thalidomide. The acetylation phenotype was predicted in all patients with N-acetyltransferase 2 genotyping. Genome DNA was tested for mutations causing an N-acetyltransferase deficiency. Homozygous individuals or those with heterozygous composites for the tested mutations were predicted slow acetylators and those with a homozygous or heterozygous genotype for an allele carrying a normal sequence at the mutation sites were predicted rapid acetylators. RESULTS: We had 7 complete responses, 1 partial response and 3 failures. Mean delay to efficacy was 7 weeks, longer for lesions involving the scalp (4 to 5 months). Six of the 8 responders were given sulfasalazine exclusively. The effect was suspensive and dose-dependent; the minimal effective dose was 1.5 g/d. Excepting light sensitization requiring discontinuation, there were no clinically significant side effects. Neutropenia occurred in one patient and moderate and transient live enzyme movements did not require treatment withdrawal. The only immunoallergic side effect (light sensitization) observed occurred in a slow acetylator. All responders except one were rapid acetylators. DISCUSSION: Salazosulfapyridine, or sulfasalazine, is composed of a derivative of 5-aminosalicylic acid and a sulfamide fraction, sulfapyridine. It is only marginally used in dermatology except for psoriasis. Its efficacy in chronic lupus erythematosus has been reported in one case. We confirmed the role of this compound in the treatment of chronic lupus erythematosus. The rare observations of induced lupus and development of antinuclear antibodies are not a contraindication, but require close regular clinical and biological surveillance. The potential risk is that possible hypersensitivity could lead to reserving sulfasalazine for severe resistant chronic lupus erythematosus after failure with antimalarials and thalidomide. Nevertheless, our study demonstrates that the slow acetylator phenotype predicts immunoallergic events, as observed by other authors, and would be a factor predicting nonresponse. If these results are confirmed by other studies, it would be possible to propose sulfasalazine as a treatment for discoid lupus erythematosus in rapid acetylators. PMID- 9740826 TI - [Pustular candidiasis in heroin addicts]. AB - INTRODUCTION: Pustular candidiasis in heroin addicts is a rare entity in dermatology. We report a case. CASE REPORT: A 29-year-old female heroin addict developed a painful pustular growth on the scalp. There was no fever. Multiple follicular pustulae measuring 2 to 3 mm were associated with hyperesthesia of the scalp and painful cervical nodes. Biopsy showed acute ostiofolliculitis with a few blastospores and mycelial filaments. Candida albicans was isolated from the pustulae and the buccal cavity. Candida serology was positive (indirect immunofluorescence 1/100, coelectrosyneresis: 4 archs). Search for other localizations and HIV serology were negative. The last injection of brown heroin had been taken 15 days earlier; lemon had been added. Treatment with flucanazole (400 mg/d) led to improvement within 48 hours. DISCUSSION: Sudden development of pustulae or nodules in pilous zones in a heroin addict should suggest the diagnosis. Outcome depends on early treatment after diagnosis and search for other localizations. Our case presented two particular aspects: ostiofollicular localization of the pustulae and a long delay (15 days) between the (presumably) last injection and the development of the lesion. Folliculitis develops almost exclusively in addicts who use brown heroin. Contamination by Candida albicans results from the lemon used to improve solubility at injection. PMID- 9740827 TI - [Perforating milia-like idiopathic calcinosis of the extremities in Down syndrome]. AB - INTRODUCTION: Several skin diseases can be seen in patients with trisomy 21. We report a case of miliary calcinosis of the extremities. CASE REPORT: A 15-year old adolescent with Down's syndrome presented small papular miliary lesions which had developed over 18 months and tended to discharge a chalk-like substance via the epidermis. Approximately 15 lesions were present on the hands and feet. Histologically, there was a well-delimited calcium deposit in the superficial dermis. There was no alteration in phosphorus/calcium metabolism. Brain CT-scan and cardiac echography did not reveal any calcifications. DISCUSSION: Miliary calcinosis cutis may not be exceptional in Down's syndrome, although only 9 observations have been reported. Preferential localizations include the hands, wrists and feet. Association with syringoma has been noted but would appear to be fortuitous. Transepidermal elimination of the calcium deposits is frequent. Pathogenic hypotheses include precipitation of calcium salts in sudation products and/or increased synthesis by fibroblasts. The association with trisomy 21 appears to be significant since only three cases have been reported in patients with normal karyotypes. This entity should be individualized as perforating milia like idiopathic calcinosis cutis of the extremities. PMID- 9740828 TI - [Perforating milia-like idiopathic calcinosis of the extremities in a patient with Down syndrome]. AB - INTRODUCTION: In 1989 a new type of calcinosis cutis has been described in association with Down's syndrome. This is the milia-like idiopathic calcinosis cutis, which is characterized by milia-like papulae generally located on the limbs (especially hands and feet) and sometimes associated with syringomas around lesions or on the eyelids. OBSERVATION: A 6 year old trisomic girl had about ten round shaped hard white-yellowish papules with a diameter of 2-3 mm on both palms of her hands. The biological balance and immunologic tests gave normal values. The histopathologic pattern was compatible with calcinosis cutis circumscripta associated with the transepidermal elimination phenomenon. Calcified sweat ducts were not observed at the von Kossa staining. Moreover, histology did not evidence any syringomas around the lesions. DISCUSSION: Our observation does not sustain the presently more spread pathogenetic interpretation, according to which eccrine ductal structures could have an active role in the formation of calcium deposits, since histology did not show any calcified eccrine ducts. Therefore, in our opinion, milia-like calcinosis associated with Down's syndrome should be classified among the idiopathic forms. PMID- 9740829 TI - [Epithelioid hemangioendothelioma: disease course in 3 cases]. AB - INTRODUCTION: Epithelioid hemangioendothelioma is an uncommon vascular neoplasm, one of intermediate-grade malignancy. Cutaneous epithelioid hemangioendothelioma is rare and often associated with multiple site involvement. CASES: Three patients with cutaneous EHE without systemic involvement in one case, with skin, liver and lung tumoral lesions in two others cases are reported. One patient is in complete remission after one year. The second patient is stable without treatment after ten years. In the third patient, alpha interferon given for one year don't produce effective results and the lesions do not progress without treatment after eight years of follow-up. DISCUSSION: Cutaneous presentation of EHE is quite variable. After the diagnosis is done, systemic involvement must be detected specially in bone, liver or lung. Metastatic spread or mulitcentric origin of the tumor are a matter of controversy. The pathobiologic behavior of EHE is not clearly recognized. Little data are available regarding the results of treatment with alpha interferon. PMID- 9740830 TI - [Acquired regressive cutaneous non-Langerhans-cell histiocytosis in an infant]. AB - INTRODUCTION: Cutaneous histiocytosis of childhood often regresses spontaneously without treatment. In some cases however, it is difficult to differentiate aggressive forms and electron microscopy and immunohistochemistry can be a valuable help. We report a case of cutaneous histiocytosis in a child which illustrates the difficulties encountered in the classification of histiocytosis. CASE REPORT: An 18-month old girl was brought to consultation with a cutaneous nodule which had developed at the age of 15 months on the labia majora. A second nodule on the chin had regressed spontaneously. Histology showed evidence of dermal histiocyte proliferation. Immunohistochemistry demonstrated is non Langerhans nature which was confirmed by electron microscopy. The clinical course was benign after surgical exeresis of the lesion on the labia majora. DISCUSSION: Different forms of histiocytosis can be classed on the basis of 4 criteria: Langerhans origin or not, acquired or congenital forms, cutaneous or visceral involvement, benign or malignant course. Four diagnosis were possible in our case: histiocytosis X, self-healing congenital histiocytosis, benign cephalic histiocytosis, juvenile xanthogranuloma. We preferred to use the descriptive term of acquired regressive cutaneous non-X histiocytosis of childhood. PMID- 9740831 TI - [Cutaneous hyalinosis: 3 cases]. AB - INTRODUCTION: The authors report three cases of "Hyalinosis cutis et mucosae"; the first and the second case concern two siblings children (brother and sister) while the third is reported on a 42-year-old woman. PRESENTATION OF CASES: The case history and the personal anamnesis, the morphology of the skin lesions, the symptoms indicating laryngeal mucous affection as well as the histopathological picture, are compatible with the "Urbach-Wiethe disease". Furthermore, in the third case, the electron microscopic findings confirm the diagnosis. DISCUSSION: The particularities of the present cases are discussed in connection with the corresponding bibliographical data. PMID- 9740832 TI - [Melanoma of soft tissues]. AB - INTRODUCTION: Soft tissue melanoma was described in 1965 by Enzinger who used the term clear-cell sarcoma. In 1983, Chung and Enzinger coined the term soft tissue melanoma due to the immunohistochemical similarity with melanoma. We report a case of this rare type of melanoma. CASE REPORT: A 59-year-old woman had pain between the first two toes for 3 years. A subcutaneous tumor was found at examination. Histologically, the tumor was composed of weakly eosinophilic cell proliferation. Protein S100 and HMB45 were positive. The cells were organized in theques. Pathology diagnosis was soft tissue melanoma. Complete remission was obtained for 3 years when several local recurrences required surgery and chemotherapy then surgery and radiotherapy. Complete remission has been achieved for 9 months. DISCUSSION: This case presented the main characteristics of soft tissue melanoma as described in a review of 209 analyzable cases reported in the literature. This tumor occurs in young subjects with no sex or race predominance. It is an ubiquitous tumor which develops in close relation with tendons and aponevroses, usually in limbs (especially feet). Pain is sometimes the revealing manifestation, but the tumor is often asymptomatic, so the volume is often important at diagnosis. Pathology examination shows rather monomorphic proliferation of cells with a clear or weakly eosinophilic cytoplasm grouped in clusters or theques separated by fibrous septa. Intracytoplasmic melanin is sometimes observed, indicating interest of protein S100 and HMB45 immunohistochemistry which is almost always positive. The principle differential diagnoses are metastasic melanoma and epithelioid sarcoma. Prognosis of soft tissue melanoma is similar to that in sarcomas with a high rate of local recurrence and metastases (lymph nodes, lungs). Mortality reaches 56 p. 100. Treatment is wide surgical exeresis. CONCLUSION: Soft tissue melanoma is a rare tumor of the melanocyte. It differs from melanoma by the population involved, its clinical expression and its prognosis which is similar to that in sarcoma. PMID- 9740833 TI - [Epilating radiotherapy: a filing document]. PMID- 9740834 TI - [Treatment of Sezary disease by leukapheresis]. PMID- 9740835 TI - [Plexiform schwannoma]. PMID- 9740836 TI - [A case for diagnosis: pyoderma vegetans]. PMID- 9740837 TI - [Lichen striatus in children and "blaschkitis" in adults]. PMID- 9740838 TI - [Cutaneous adverse effects of mesotherapy]. PMID- 9740839 TI - [Transgenic mice: a model for the study of hereditary cutaneous bullous diseases]. PMID- 9740840 TI - [Question of the month: should coal-tar be forbidden?]. PMID- 9740841 TI - [Modern dermatologic treatments: a few comments]. PMID- 9740842 TI - [Isotretinoin (Roaccutane) in women of childbearing age: failure of following prescription guidelines]. AB - BACKGROUND: Despite prominent warnings, pregnancies continue to be reported in women exposed to isotretinoin. PATIENTS AND METHODS: We report results of the analysis of 318 questions asked to pharacovigilance structures in France from 1987 to 1995 because of an exposition to isotretinoin during the risk period and of a prospective inquiry concerning isotretinoin prescription in women conducted among pharmacists. RESULTS: These 318 pregnancies began during the month after Roaccutane withdrawal (n = 104, 33 p. 100), during Roaccutane treatment (n = 163, 51 p. 100) or before Roaccutane treatment (n = 51, 16 p. 100). Of the 267 women with pregnancies conceived during treatment with isotretinoin (n = 104) or during the month after its discontinuation (n = 163), contraception was not prescribed in 28 (15 p. 100) or prescribed but with poor compliance in 109 (60 p. 100). Pregnancy was terminated voluntarily in 199 women (81 p. 100). In the 173 women who were interviewed in pharmacies, 49 (28 p. 100) did not use contraception and among them contraception was prescribed in only 59 p. 100. Only 14 p. 100 had received full information about isotretinoin and pregnancy. The teratogenic effects of isotretinoin were known by 98 p. 100 of the women and the need of contraception during treatment and for one month after discontinuation by 70 p. 100. DISCUSSION: Insufficient compliance with warnings is the main reason for pregnancies in women receiving isotretinoin therapy. A pregnancy prevention program is needed before prescription to ensure comprehension and to obtain informed consent of patients. PMID- 9740843 TI - [Ketoprofen-induced contact photosensitivity disorders: 5 cases]. AB - BACKGROUND: Nonsteroidal antiinflammatory drugs (NSAID) are widely used in topical applications for benign diseases. Adverse skin reactions include contact eczema and photocontact dermatitis. Among the NSAID used in topical applications, arylpropionic derivatives, notably ketoprofen, are frequently implicated. CASE REPORTS: We observed 5 patients who developed eczema lesions after application of Ketum, a gel containing ketoprofen used on healthy skin after exposure to sunlight. Photoallergy explorations evidenced positive photopatch-tests for ketoprofen with UVA and total light. The anamnesis suggested a photoallergic mechanism which was confirmed by histological examination of the biopsy of a UVA positive photopatch-test and by negative photopatch-tests in 10 healthy controls. DISCUSSION: The photosensitizing potential of ketoprofen in the UVA spectrum is well known. Although the number of adverse reactions is quite small compared with widespread use, physicians should be aware of this photosensitivity and report all cases to the pharmacovigilance center. PMID- 9740844 TI - [Unilesional plaque-type mycosis fungoides: 3 cases]. AB - BACKGROUND: The clinical course of unilesional plaques with a histological diagnosis of mycosis fungoides is generally benign. This uncommon situation raises the question of nosology among epidermotropic T-cell proliferations. CASE REPORT: Two women (age 65 and 70 years, cases 1 and 2) and one man (age 66 years, case 3) each had a well-limited unique oval-shaped plaque measuring 1 to 3 cm in diameter for several months. The localization of the erythematous, squamous plaques with minimal infiltration and little or no pruritus remained unchanged in the dorsal area. Histology reported mycosis fungoides: dense superficial dermal infiltration in band attached to the epidermis with a clear lower limit, formed by small and medium-sized atypical lymphocytes, sometimes with cerebriform nuclei. Epidermotropism on isolated cells or theques was noted. There was no spongiosis. Immunolabeling demonstrated predominance of CD4+ cells and tumoral infiltration uptake of anti-V beta 8 monoclonal antibodies in the two cases examined. The lesion totally regressed in case 1 after local application of carmustine with no recurrence after 5 years. Patient 2 declined treatment and no change was observed 5 years later. Surgical exeresis was performed in case 3 without recurrence 3 years later. DISCUSSION: No histological feature or T-cell immunophenotype distinguishes this clinical entity from classical mycosis fungoides. As no other long-term lesions developed, our three cases would argue for a benign course of what is probably a variety of mycosis fungoides. Unilesional plaque mycosis fungoides must be lymphomatoid contact dermatitis. Woringer-Kolopp disease differs by its nearly exclusive epidermotropic distribution. PMID- 9740845 TI - [Collagenolysis above eccrine spiradenoma]. AB - BACKGROUND: The casual observation of a peritumoral edema with collagenolysis in a case of eccrine spiradenoma led us to search for similar lesions in a retrospective series of 50 spiradenomas. MATERIAL AND METHODS: Among 50 cases of eccrine spiradenomas, 36 excised with the overlying epidermis and the surrounding dermis were finally available for a conventional histologie study. RESULTS: In 14 cases, i.e. 39p. 100, we observed an edema above the tumour with thin and sparse collagen fibers and less altered elastic fibers. The clinical presentation of these 14 cases was not different from the general presentation of eccrine spiradenomas, but no precise photographical or clinical data were available. DISCUSSION: The cause of this epitumoral collagenolysis is unknown. It seems unrelated to the clear perivascular spaces often observed within the tumour lobules. It is of no help for diagnosis and may be considered as an unexplained curiosity like the elastolysis and the anetoderma seen above some pilomatricomas. PMID- 9740846 TI - [Coronary involvement in systemic lupus erythematosus]. AB - BACKGROUND: Coronary artery disease is an uncommon event in lupus erythematosus. The mechanisms responsible for coronary occlusion are probably complex and intermixed. We report three patients with lupus erythematosus and antiphospholipid antibodies who had coronary artery disease diagnosed with coronary angiogram. OBSERVATION: Coronary artery disease occurred in three young patients aged from 21 to 35 years 3 to 11 years after the onset of lupus. They all had antiphospholipid antibodies. They had been treated with corticosteroids for 6 to 36 months. Two of them were smokers. Angiograms showed coronary occlusion two patients while the third one had probable myocardial microvasculopathy. The lupus was quiescent in all cases when coronary artery disease occurred. DISCUSSION: Antiphospholipid antibodies associated with smoking may be involved in the pathogenesis of coronary artery disease in these 3 patients. PMID- 9740847 TI - [Scleredema, acanthosis nigricans and IgA/Kappa multiple myeloma]. AB - BACKGROUND: Scleredema is an uncommon disease of unknown origin. Characteristic thick skin with symmetrical diffuse induration develops. The infiltration begins on the face and neck then extends to the root of the upper limbs and trunk. There are three clinical types of scleredema. The first is preceded by an upper airway infection and progresses rapidly before regressing spontaneously within a few months. The second type is associated with chronic diabetes. The third type is associated with monoclonal gammapathy, rarely of myelomatous type, and develops insidiously. Acanthosis nigricans can be a paraneoplastic syndrome, often associated with a gastrointestinal cancer. Few cases associating scleredema and acanathosis nigricans have been reported. CASE REPORT: A 56-year old woman had developed scleredema over the last 6 years when acanthosis nigricans appeared together with IgA kappa multiple myeloma. Treatment with melphalan and prednisolone was effective against the myeloma as well as the scleredema and acanthosis nigricans. DISCUSSION: Only five cases of associated scleredema and multiple myeloma have been reported, four with kappa IgG myeloma and one with IgA myeloma. An association between acanthosis nigricans and sclerederma could be coincidental although the fact that the different manifestations regressed together after the myeloma treatment would suggest some relationship between these three diseases. PMID- 9740848 TI - [Ofuji's eosinophilic pustular folliculitis. Efficacy of acitretin]. AB - BACKGROUND: We report the first case of eosinophilic pustular folliculitis (Ofuji's disease) which was successfully treated with acitretin. CASE REPORT: A 50-year old women (HIV negative) had developed over 3 months an erythematopapulous plaque under the left orbit. The clinical and histological diagnosis was eosinophilic pustular folliculitis. Successive treatment with cetirizine then indomethacin was ineffective. Acitretin (0.5 mg/kg/d) was then started and led to cure within 1 month. Six weeks after the patient spontaneously stopped the treatment, the lesion recurred at the same localization. Further treatment with isotretinoin (0.5 mg/kg/d) was then given but did not alter progression of the lesion. Acitretin was then reintroduced at the same dose and again produced rapid cure. Acitretin was then tapered off to 10 mg/d then maintained at this dose as lesions reappeared with further dose reduction. DISCUSSION: It is difficult to treat eosinophilic pustular folliculitis because of the random nature of response to different drugs. General corticosteroids, dapsone and indomethacin are classically proposed but with variable success. Isotretinoin is proposed on the hypothesis of a link with sebaceous secretion, but results have been contradictory. This drug was ineffective in our case. Acitretin did however provide very rapid improvement with an efficacy confirmed at reintroduction. This retinoid which does not have the specific action of isotretinoin could affect follicular keratinocytes which have been shown to be activated in this chronic skin disease. PMID- 9740849 TI - [Macrophage activation syndrome disclosing leukemic transformation of mycosis fungoides]. AB - BACKGROUND: Mycosis fungoides can mimic pigmented purpuric dermatitis. We report such a case which progressed to peripheral T-cell lymphoma; progression was revealed by reactive hemophagocytic syndrome (RHS). CASE REPORT: A 65-year old male patient was hospitalized for a pigmented and purpuric eruption. The skin lesions appeared 2 years earlier and at that time biopsy had shown pigmented and purpuric dermatitis. One month before hospitalization, general signs appeared. On admission, he had papular and purpuric rash, mainly on the trunk, hepatosplenomegaly, enlarged axillar and inguinal lymph nodes, and fever at 38.2 degrees. A skin biopsy showed histologic changes typical of mycosis fungoides. He also had bicytopenia, hepatitis, and increased triglyceride and ferritin levels suggesting RHS which was proved by means of bone marrow biopsy. These tests also evidenced peripheral T-cell lymphoma. The patient was treated with two courses of chemotherapy (CHOP) but the disease progressed and he deceased. DISCUSSION: Mycosis fungoides can occasionally begin with an eruption very closely resembling pigmented purpuric dermatitis. Therefore, repeated biopsies should be done in case of widespread permanent pigmented purpuric dermatitis of no apparent origin. RHS is a life-threatening disease. The diagnosis should be suspected in any cytopenic patient with fever, increased triglyceride levels and abnormal liver tests. A search for an etiology must then be undertaken a prompt treatment is needed. PMID- 9740850 TI - [Acute generalized exanthematic pustulosis after intake of clozapine (leponex). First case] case]. AB - BACKGROUND: Acute generalized exanthematic pustulosis is a severe adverse drug reaction which occurs after taking antibiotics. Rare cases implicating psychotrops have been observed. CASE REPORT: A 71-year old women with schizophrenia was given closapine for six weeks when she developed an erythematopustular skin reaction and fever typical of acute generalized exanthematic pustulosis. The skin disease regressed one week after withdrawing clozapine. DISCUSSION: This is the first case of acute generalized exanthematic pustulosis observed after taking the neuroleptic drug, clozapine, used in severe schizophrenia. PMID- 9740851 TI - [Encephalocraniocutaneous lipomatosis]. AB - BACKGROUND: Encephalocraniocutaneous lipomatosis (ECL) involving the scalp and cerebellum was observed without asymptomatic expression. CASE REPORT: A seven month-old infant presented with two soft subcutaneous hairless tumors of the scalp without any associated clinical anomaly. Neuroradiology explorations (radiography, CT-scan and MRI) showed a lipoma in the cerebellum linked with a occipital cutaneous lipoma through a bone defect. At the age of 3 years, the child remains healthy. DISCUSSION: ECL is a rare neurocutaneous disorder that consists of skin lipomas associated with various cerebral anomalies. ECL may occur as a circumscribed form of the Proteus syndrome, since a few ECL patients have associated manifestations of proteus syndrome as well as localized hypertrophy. However, minor forms of ECL are possible and may be compatible with normal life. PMID- 9740852 TI - [Chloroquine-induced achromotrichia. An ultrastructural study]. AB - BACKGROUND: Electron microscopy examination of scalp biopsies from a patient with chloroquine achromotrichia gave elements concerning the pathogenesis of chloroquine-induced achromotrichia. CASE REPORT: A 21-year old light brown-haired patient developed achromotrichia after four months of treatment with chloroquine for subacute lupus erythematosus. Hair bleaching completely regressed 5 months after discontinuing chloroquine despite replacement with hydroxychloroquine. During the achromatrichia phase, many ultrastructural anomalies were observed in the hair root melanocytes: the nuclei were small and densified, and there was an accumulation of immature melanosomes in the cytoplasm; these melanosomes, mainly in stage II, were rarely transferred to keratinocytes. After recovery from the achromotrichia, melanocytes displayed a normal aspect. DISCUSSION: Pathophysiological disturbances leading to chloroquine induced achromotrichia are still unclear. The ultrastructural study of hair follicles in our patient show that under chloroquine action melanocytes become unable to perform complete melanin synthesis and to produce normally melanized melanosomes which may be transferred to keratinocytes. Non-melanized or poorly melanized melanosomes accumulate in the melanocytes which finally become inactive cells. These findings suggest that achromotrichia is induced by a toxic effect of chloroquine on the melanocyte. PMID- 9740853 TI - [Risk of viral infection and CO2 laser]. PMID- 9740854 TI - [Melanoma in Maghreb]. PMID- 9740855 TI - [A case for diagnosis: Fox-Fordyce disease]. PMID- 9740856 TI - [A case for diagnosis: Hallopeau-type pemphigus vegetans]. PMID- 9740857 TI - [A look on research: gene p53 and cutaneous carcinoma]. PMID- 9740858 TI - [Treatment of superficial skin infections caused by Staphylococcus pyogenes]. PMID- 9740859 TI - [New allergens in cosmetics. Cosmetovigilance]. PMID- 9740860 TI - [Question of the month: could methodological steps and legal procedure of new therapeutic approval be bypassed?]. PMID- 9740861 TI - [Hypereosinophilic syndrome: the dermatologist's point of view]. PMID- 9740862 TI - [Should tinea capitis in schools be reported?]. PMID- 9740863 TI - [Staphylococcus aureus septicemia producing Panton-Valentine leukocidin. 3 cases]. AB - BACKGROUND: A strong association has been observed between furuncles and Panton Valentine leukocidin-producing Staphylococcus aureus. CASE REPORTS: Within one year, we cared for three men at the Cayenne hospital who had Staphylococcus aureus septicemia with severe pleuropulmonary involvement originating from furuncular lesions. The Staphylococcus aureus strains isolated from the skin lesions and from blood cultures produced Panton-Valentine leukocidin. CONCLUSION: These cases demonstrate the gravity of S. aureus septicemia in young patients with furunculosis. These cases are the first reported with severe S. aureus infections associated with Panton-Valentine leukocidin producing strains. PMID- 9740864 TI - [Rilmenidine in rosacea: a double-blind study versus placebo]. AB - INTRODUCTION: The usual treatments of rosacea (cyclines and metronidazole) are mainly effective on reducing the number of papules and pustules. Clonidine only was employed in order to treat flushes observed in rosacea. Rilmenidine is a central hypotensive drug which acts more specifically than clonidine on imidazoline receptors and which has no sedative side effects. The purpose of this study was to evaluate the efficacy of rilmenidine 1 mg/d in the treatment of rosacea. PATIENTS AND METHODS: A total of 41 patients suffering from typical rosacea were selected in this randomised double blind study rilmenidine versus placebo. The study comprised an 1-month observation period without treatment followed by 3 months of treatment. The major assessment criteria was the proportion of responders at the end of the treatment period. Responders were defined as patients showing a decrease of more than 50 p. 100 in the count of papules and pustules. Minor criterias were the variation of the number of flushes, self-evaluated by the patient and the variation of the redness of the face, noted by the investigator on a scale from 0 to 5. RESULTS: Fifteen patients treated by rilmenidine (R) and 19 patients receiving the placebo (P) were evaluated. The proportion of responders was 69.2 p. 100 in group R and 57.1 p. 100 in group P (p = 0.69). The variations of the number of papules and pustules and of the redness of the face were not significantly different in the two groups. The decrease in the number of flushes was higher in group R (around -13) than in group P (around -5), but the difference was not really significant (p = 0.076). Arterial pressure decreased in 3 patients in group R and in 2 patients in group P. Minor side effects were noted in a similar proportion of patients in the two groups. DISCUSSION: Rilmenidine is not efficient in reducing the number of papules and pustules but could decrease the number of flushes. Nevertheless, many patients were lost for follow-up and because of a major placebo effect, the conclusions of this study are not strong enough. Another study including more patients and using evaluation criteria based on the vascular components of rosacea could perhaps confirm this hypothesis. PMID- 9740865 TI - [Acne, hyperandrogenism and oral isotretinoin resistance. 23 cases. Therapeutic implications]. AB - BACKGROUND: We earlier demonstrated that oral isotretinoin can be associated with hyperandrogenism in women with acne. The aim of this study was to evaluate the causal relationships of the different etiologies in case of unsuccessful treatment. PATIENTS AND METHODS: The study group included 120 patients with late onset acne resistant to different treatment and signs of hyperandrogenism. A complete hormone work-up was obtained in all patients. There was a group of 23 patients who failed to respond to isotretinoin and 97 patients in the control group. Unsuccessful treatment was defined as persistance of grade 2 lesions after a mean cumulative dose of 166 mg/kg isotretinoin. RESULTS: In the non-responders to isotretinoin, hyperandrogenism was observed in 22 out of 23 cases: pituitary (n = 2), adrenal (n = 5), ovarian (n = 13), peripheral (n = 2). In the control group, hyperandrogenism was found in 89 out of 97 patients: pituitary (n = 6), adrenal (n = 45), ovarian (n = 33), peripheral (n = 5). The distribution of two etiologies, ovary and adrenal, demonstrated a significant difference between isotretinoin non-responders and controls, the former having a higher frequency of ovarian hyperandrogenism. DISCUSSION: These findings confirm that untreated hyperandrogenism, particularly ovarian hyperandrogenism, is a source of unsuccessful treatment with oral isotretinoin. PMID- 9740866 TI - [Epidemiology of Microsporum langeronii tinea capitis in the Paris suburban area. Results of 2 school and familial surveys]. AB - INTRODUCTION: One epidemiologic survey was carried out in two children communities, following detection of several cases of Microsporum langeronii tinea capitis. PATIENTS AND METHODS: In one case, 3 year-old children from a day nursery were contaminated by a child originating from France, who had been previously infected by contact with a friend originating from Ivory Coast. In the second case, lesions were diagnosed in a nursery-school in African children born in France. These were treated before epidemic progression into the school. RESULTS: Our study confirms data from the literature concerning the risk of contamination by Microsporum langeronii, with a familial contamination being more frequent than scholar one. DISCUSSION: The interest of our study was the rapid starting of the epidemiologic survey after first case diagnosis (one to two months) and the treatment of all the patients without scholar eviction. Treatment of all affected patients as well as "asymptomatic carriers" leaded to the arrest to the epidemy. No case of relapse was noted during the following year. PMID- 9740867 TI - [Spironolactone-induced pemphigoid]. AB - BACKGROUND: Sporadic observations would suggest that certain drugs play a role in the development of pemphigoid. A recent case-control study on long-term drug use associated with pemphigoid was unable to confirm the suspected role of these drugs, but did demonstrate a significant association between the development of pemphigoid and use of spironolactone. CASE REPORT: An 82-year-old patient had bullous erythematous lesions on the left thigh suggestive of pemphigoid. Histology and direct cutaneous immunofluorescence confirmed the diagnosis. The patient had been taking alimemazine, adrafinil, flunarizine, spironolactone and furosemide as long-term treatment. Spironolactone alone was withdrawn. The cutaneous lesions regressed and have not recurred during the 18-month follow-up. DISCUSSION: The causal effect of spironolactone in this case of pemphigoid is quite plausible. This hypothesis is favored by the agreement with epidemiological data. Other observations are however required before the causal role of spironolactone in development of pemphigoid can be confirmed. PMID- 9740868 TI - [Erosive lichen of the scalp]. AB - BACKGROUND: We report an exceptional clinical presentation of erosive lichen planus of the scalp associated with hepatitis C and idiopathic CD4 lymphocytopenia. CASE REPORT: A 90-year-old woman was hospitalized in May 1995 for erosive dermatosis of the scalp and alopecia, associated with ungual lesions. Histology of the scalp lesions demonstrated lichen and the serology tests were positive for hepatitis C. The patient also had severe CD4 lymphocytopenia (290/mm3). The lesions regressed with clobetasol and occlusion and growth of scalp hair turned. DISCUSSION: Erosive lichen planus of the scalp has, to our knowledge, not been reported previously. The causal effect of drugs taken by this patient (tetrazepam and clorazepam) was not retained. Seropositivity for hepatitis C (with no evidence of active disease) was however probably not fortuitous. The association with CD4 lymphocytopenia merits discussion. PMID- 9740869 TI - [Palmoplantar filiform parakeratotic hyperkeratosis and digestive adenocarcinoma]. AB - INTRODUCTION: There are very few observations of filiform palmo-plantar hyperkeratosis reported. Nevertheless it's worth knowing this entity for his potential association with a visceral neoplasia. CASE REPORT: We report the first case of filiform palmo-plantar hyperkeratosis associated with a digestive adenocarcinoma and a polycystic kidney disease. DISCUSSION: After a review of palmar and plantar filiform hyperkeratosis in the literature, we will discuss the possible association with neoplasia or other pathologies. This pathology requires a strict clinical and paraclinical follow-up. PMID- 9740870 TI - [BCG-induced mycobacterium infection induced by alternative medicine]. AB - INTRODUCTION: Mesotherapy is an alternative medical process defined by the intradermal injection of small amounts of pharmacological substances. It has been associated with the development of mycobacterial infections. CASE REPORT: A 80 year-old woman presented with a Mycobacterium bovis infection of the face following mesotherapy. Genome analysis of the mycobacterial strain isolated from a skin lesion using pulse-field gel electrophoresis demonstrated the presence of a vaccinal strain identical to the one employed by the same physician to vaccinate a child two hours before. DISCUSSION: Given the severity of mycobacterial infections following mesotherapy and given the lack of evidence about its efficacy, mesotherapy should not be performed on the face. Standardisation of aseptic measures in the daily medical practice could prevent such infectious complications. PMID- 9740871 TI - [Epidermal hamartoma and blaschkitis in an adult]. AB - INTRODUCTION: Many inherited skin diseases and nevus malformations are distributed according to the Blaschko's lines. Some inflammatory skin diseases are also distributed according to these lines. OBSERVATION: In 1989 a 49-year-old woman presented with an epidermal nevus affecting the left hand and leg from the infancy. In 1994, inflammatory, linear papules occurred on the left leg, obscuring and continuing the previous lesions, on the left thigh and on the left clavicular region. On light microscopy, spongiosis of the epidermis and an infiltrate of lymphocytes and histiocytes in the superficial dermis were shown. Subintrant crops of new inflammatory lesions occurred for a period of two years, whereas the nevus lesions of the left hand persisted unchanged. These findings led to the final diagnosis of blaschkitis associated to epidermal nevus on the same Blaschko's lines. DISCUSSION: The distribution of a skin disease according to the Blaschko's lines underlines the presence in the affected skin of a mutant clone, with a different genetic material as compared with the normal skin. An early mutation giving raise to a mutant clone and affecting the left hemibody could be hypothesized in our case. The mutation probably involved a pleiotropic gene. The latter initially was responsible for thickening of the skin. Moreover, the mutation was also responsible for a particular proneness towards an exogenous factors, able to induce a persistent inflammatory skin eruption following the same lines. PMID- 9740872 TI - [Chronic edema, monoclonal dysglobulinemia and profuse telangiectasia: a distinct entity?]. AB - INTRODUCTION: Monoclonal gammopathy and edema are features encountered during several diseases, especially systemic capillary leak syndrome. The diagnoses of POEMS syndrome, edematous systemic scleroderma and a fortuitous association may be also discussed. We report the cases of two patients which did not fulfill the criteria for such diagnoses. CASE REPORT: Although the 2 cases share some discrepancies, they have also similar and particular features: association of chronic edema, monoclonal gammopathy and profuse and acquired telangiectasias. DISCUSSION: The meaning of these cases remains to be clarified. It might be an entity close to the systemic capillary leak syndrome but characterized by the chronicity of edema and by a distinct cutaneous sign, the occurrence of numerous telangiectasias. These cases emphasize that the acquired and profuse telangiectasias belong to the wide range of cutaneous abnormalities which may be associated with monoclonal paraproteinemia. PMID- 9740873 TI - [Topical neomycin: risks and benefits. Plea for withdrawal]. PMID- 9740874 TI - [Diffusion of minocycline in comedons of patients with acne vulgaris]. PMID- 9740875 TI - [Autochtonous leprosy]. PMID- 9740876 TI - [Treatment of rosacea. Clonidine (0.075 mg per day) versus placebo (initial results)]. PMID- 9740877 TI - [A case for diagnosis: eruptive collagenoma]. PMID- 9740878 TI - [Epidemiology of nevus]. PMID- 9740879 TI - [Staphylococcal leukotoxins]. PMID- 9740880 TI - [Question of the month: should AIDS serodiagnosis be requested in every patient with a common form of dermatosis?]. PMID- 9740881 TI - Communicating the evidence. PMID- 9740882 TI - Pattern recognition: a complex process. PMID- 9740883 TI - Weaning from mechanical ventilation: patterns in young children recovering from acute hypoxemic respiratory failure. AB - OBJECTIVE: The purpose of the study was to describe the patterns of weaning from mechanical ventilation in young children recovering from acute hypoxemic respiratory failure. METHODS: Decision-making rules on progressive weaning were developed and applied to existing data on 82 patients 2 weeks to 6 years old in the Pediatric Acute Respiratory Distress Syndrome Data Set. RESULTS: Three patterns of weaning progress were detected: sprint, consistent, and inconsistent. Length of ventilation and weaning progressively increased from the sprint, to the consistent, to the inconsistent subset. Patients in the inconsistent subset were most likely to have a systemic (sepsis or shock) trigger of acute respiratory distress syndrome and to be rated as having at least moderate disability at discharge. Hypothesis-generating univariate and then multivariate logistic regression analyses indicated that patients who experienced more days of mechanical ventilation before the start of weaning and who had a higher oxygenation index during the weaning process were most likely to have an inconsistent pattern of weaning. CONCLUSION: Patterns of weaning are discernible in a population of young children and indicate a subset at risk for inconsistent weaning. Knowing the patterns of weaning may help clinicians anticipate, perhaps plot, and then modulate a patient's weaning trajectory. PMID- 9740884 TI - Comparison of 2 methods of measuring the QT interval. AB - BACKGROUND: Prolonged cardiac repolarization is associated with ventricular tachycardia and sudden cardiac death. Repolarization, represented by the QT interval, is usually measured on a 12-lead ECG recording. Measurements of the interval on bedside monitor ECG recordings have not been compared quantitatively with measurements on 12-lead ECG recordings. OBJECTIVE: To determine if QT intervals and QTc values obtained by using monitor recordings are as accurate as those obtained by using 12-lead ECG recordings. METHODS: For each of 50 subjects, 2 ECG recordings were obtained, 1 with a 12-lead ECG and 1 with the bedside monitor, and QT intervals were measured manually. The QT intervals on each type of recording were compared on a lead-by-lead basis, the maximum QT interval and the QTc maximum determined with each method were compared, and the "best single leads" for determining the QTc were ascertained for each method. RESULTS: QT intervals, on a lead-by-lead basis; maximum QT intervals; and QTc maximum values measured on the monitor recordings were consistently longer than those measured on the 12-lead ECG recordings. When the monitor ECG leads I or II and the 12-lead ECG QTc maximum were examined for simple agreement by using 460 milliseconds as a cutoff, agreement was found in 82% to 84% of the sample, and false negatives were 12% and 8%, respectively. CONCLUSION: Recordings from leads I or II on the bedside ECG monitor should be used to measure the QT interval. Once prolonged QT values are detected, recordings obtained with a 12-lead ECG can be used to confirm the analysis. PMID- 9740885 TI - 12-lead ST-segment monitoring vs single-lead maximum ST-segment monitoring for detecting ongoing ischemia in patients with unstable coronary syndromes. AB - BACKGROUND: 12-lead ECG monitoring of the ST segment is more sensitive than patients' symptoms for detecting ischemia after thrombolytic therapy or catheter based interventions, but it is unclear whether monitoring of the single lead showing maximum ST deviation would be as efficacious. OBJECTIVE: To determine whether monitoring all 12 ECG leads for changes in the ST segment is necessary to detect ongoing ischemia in patients with unstable coronary syndromes. METHODS: Continuous 12-lead ST segment monitoring was performed in 422 patients from the onset of myocardial infarction or during balloon inflation in catheter-based interventions until the patient's discharge from the cardiac care unit. Computer assisted techniques were used to determine (1) which lead showed the maximum ST deviation at the onset of myocardial infarction or during balloon inflation and (2) what proportion of later ischemic events were associated with ST deviation in this lead. RESULTS: The lead with the maximum ST deviation could be determined in 312 patients (74%). The remaining 110 (26%) had non-Q wave infarction without ST deviation or no ST changes during balloon inflation. During 18,394 hours of 12 lead ST monitoring, 118 (28%) of the 312 patients had a total of 463 ischemic events, 80% of which were silent. Of 377 ischemic events in which a maximum ST lead was detected, 159 (42%) did not show ST deviation in this lead (sensitivity, 58%; 95% CI, 53%-63%). Routine monitoring of leads V1 and II showed ST deviation in only 152 of the 463 events (sensitivity, 33%; 95% CI, 29%-37%). CONCLUSIONS: Monitoring of all 12 ECG leads for changes in the ST segment is necessary to detect ongoing ischemia in patients with unstable coronary syndromes. PMID- 9740886 TI - Opioid withdrawal in neonates after continuous infusions of morphine or fentanyl during extracorporeal membrane oxygenation. AB - BACKGROUND: Complications of opioid analgesia include tolerance and withdrawal. OBJECTIVES: To determine the effects of morphine and fentanyl on the prevalence of withdrawal after extracorporeal membrane oxygenation. METHODS: Two groups of neonates were compared during and after extracorporeal membrane oxygenation: a prospective group receiving a continuous infusion of morphine for analgesia and sedation and a retrospective group who had received a continuous infusion of fentanyl. RESULTS: Neonates receiving morphine required significantly less supplemental analgesia (P < .001) than did neonates who had received fentanyl and had a significantly lower prevalence of withdrawal after the therapy (P = .01). Neonates receiving morphine were discharged from the hospital a mean of 9.6 days sooner (P = .01) than neonates who had received fentanyl. CONCLUSIONS: Morphine may offer marked advantages over fentanyl for providing continuous analgesia and sedation in neonates. PMID- 9740887 TI - Clinical evaluation of noninvasive monitoring of oxygen saturation in critically ill patients. AB - OBJECTIVE: To examine the effect of abnormal cardiac index on the accuracy of measurement of oxygen saturation by pulse oximetry. METHODS: Forty-six patients (mean age, 49 years) in a 9-bed medical ICU were studied. Measurements of oxygen saturation obtained with pulse oximeters and with a functional cooximeter were collected at baseline and 4, 8, 16, 24, 32, 40, and 48 hours later. Hemodynamic and cardiopulmonary parameters were recorded. RESULTS: The Bland-Altman technique yielded upper and lower limits of agreement of 2.53% and -7.11%. Most (95.7%) of the differences between the measurements of oxygen saturation obtained with the 2 methods were within these limits, although some of these differences may be clinically unacceptable. The bias was -2.29%, and the precision was 2.41%. The clinical conditions associated with inaccurate tracking of saturation by pulse oximetry across the range of actual arterial oxygen saturation values were abnormal cardiac index, partial pressure of carbon dioxide, heart rate, and pulmonary capillary wedge pressure. CONCLUSIONS: In patients with abnormal cardiac index, the pulse oximeter measurements exceeded the actual oxygen saturation by up to 7%. Pending prospective studies, clinicians should be aware that when certain cardiopulmonary parameters are abnormal, the margin of error in measurements of oxygen saturation obtained with a pulse oximeter may be greater than when those parameters are normal. PMID- 9740888 TI - Can measurement of mixed venous oxygen saturation replace measurement of cardiac output in patients with advanced heart failure? AB - BACKGROUND: Nursing care of patients with advanced heart failure with low ejection fraction requires strict management of IV fluids. Measurement of mixed venous oxygen saturation offers advantages over measurement of cardiac output because no administration of fluid is required and data are obtained continuously. OBJECTIVES: To determine the relationship between mixed venous oxygen saturation and cardiac output in patients with advanced heart failure who have low ejection fraction and to determine if use of vasoactive medications alters the relationship between mixed venous oxygen saturation and cardiac output. METHODS: Simultaneously obtained measurements of mixed venous oxygen saturation and cardiac output were compared in 42 patients with advanced heart failure with ejection fractions of 30% or less (mean, 19.5%). RESULTS: Correlation between mixed venous oxygen saturation and cardiac output was r = 0.54 (P < .001). For subjects not receiving vasoactive medications (n = 28), r = 0.52 (P = .004); for those receiving vasoactive medications (n = 14), r = 0.57 (P = .03). CONCLUSIONS: Similar correlations in the groups receiving and not receiving vasoactive medications suggest that even with pharmacological support, changes in mixed venous oxygen saturation may not be reflected by concomitant changes in cardiac output. Measurement of mixed venous oxygen saturation should not replace measurement of cardiac output for clinical decision making in patients with advanced heart failure with low ejection fraction. PMID- 9740889 TI - Impact of family demands and family strengths and capabilities on family well being and adaptation after critical injury. AB - BACKGROUND: Increases in demands on patients' family members that are not reduced by family strengths may contribute to decreases in family adaptation and complicate patients' recovery after trauma. The purpose of this study was to examine family demands (prior stressors and severity of patients' injuries) and family strengths and capabilities (hardiness, resources, coping, and problem solving communication) associated with outcomes of family well-being and adaptation. METHODS: A multivariate, descriptive design based on the Resiliency Model of Family Stress was used. A convenience sample of family members (N = 51) of adult patients participated within the first 2 days of critical injury. Family demands were measured with the Family Inventory of Life Events and Changes and the Acute Physiology, Age, and Chronic Health Evaluation III. Family strengths were measured with the Family Hardiness Index, Family Inventory of Resources for Management, Family Crisis Oriented Personal Evaluation Scale, and Family Problem Solving Communication Index. Family adaptation outcomes were measured with the Family Well Being Index and Family Adaptation Scale. RESULTS: Increases in family demands were significantly related to decreases in family strengths and family adaptation. Family demands scores accounted for 40% of the variance in family well-being scores. The only significant family strength variable influencing family adaptation was problem-solving communication. CONCLUSIONS: Increases in family demands seem to be an important indicator of the amount of assistance a family may need. Interventions that help mobilize family strengths, such as problem-solving communication, may be effective in promoting the adaptation of families of critically injured patients. PMID- 9740890 TI - Successful medical management of a patient with an anomalous right coronary artery who declined surgery. PMID- 9740891 TI - Therapy for hyperlipidemia when it is the only risk factor--fact or fiction? PMID- 9740892 TI - Nurses' knowledge of advance directives. PMID- 9740893 TI - Prospective pricing system for nursing facilities begins. PMID- 9740894 TI - Panel cites policies for preventing medication misuse in the elderly. PMID- 9740895 TI - Monoclonal antibody approved for RSV disease prevention. PMID- 9740896 TI - Oral migraine drug approved for marketing. PMID- 9740897 TI - Overhaul of military pharmacy system recommended. PMID- 9740898 TI - Core formulary issued for military treatment facilities. PMID- 9740899 TI - Survey of pharmacy benefit trends released. PMID- 9740900 TI - Obesity guidelines prescribe limited role for weight-loss drugs. PMID- 9740901 TI - The quest for quality measurement. PMID- 9740902 TI - Harvey A. K. Whitney Lecture. The patient is waiting. PMID- 9740904 TI - Rededicating ourselves to service: becoming the 'visible' ingredient in health care. PMID- 9740905 TI - Bold in our C.A.R.E. (commitment, advocacy, relationships, education). PMID- 9740906 TI - Diversification continues to be key to success. PMID- 9740907 TI - Leading health-system pharmacy into the next century. PMID- 9740908 TI - ASHP Board of Directors, councils, committees, and staff. PMID- 9740910 TI - ASHP strategic-planning report for 1998-99. PMID- 9740909 TI - Officers of ASHP affiliated state societies. PMID- 9740911 TI - ASHP statement on the pharmacist's role in substance abuse prevention, education, and assistance. PMID- 9740912 TI - ASHP statement on the pharmacist's role in infection control. PMID- 9740913 TI - ASHP statement on the pharmacist's role in clinical pharmacokinetic monitoring. PMID- 9740914 TI - Two antiretroviral drugs likely to be confused. PMID- 9740915 TI - Endoscopically guided cultures in chronic sinusitis. AB - In chronic sinusitis, culture-directed antibiotics are often recommended as a cornerstone of treatment. The significance of Gram-negative rods (GNRs), coagulase-negative Staphylococci (SCN), and Staphylococcus aureus has been controversial. In an effort to determine host factors which correlate with culture results, 507 endoscopically-guided cultures are reviewed from 265 patients. A history of asthma, allergic rhinitis, prior sinus surgery, and the concurrent use of antibiotics, steroids, and irrigations were some of the host factors compared by X2. The results were compared to a control group of 50 cultures from healthy volunteers. SCN, S. aureus, P. aeruginosa, and Streptococcus were the most common isolates. GNRs were present in 27% of cultures and were more common in patients who had prior sinus surgery or were using irrigations. P. aeruginosa was more common in patients taking systemic steroids. SCN occurred with the same incidence in patients and control subjects but was more prevalent in cultures obtained intraoperatively and in patients taking systemic steroids. No identifiable host factor was associated with S. aureus. S. aureus occurred at similar rates in patients and control subjects but grew heavily in patients and exhibited only light growth in controls. Topical nasal steroids appear to have no statistically significant effect on bacterial cultures. Findings from this study further our understanding of chronic sinusitis and may help guide practitioners in the treatment of this disease. PMID- 9740916 TI - Antimicrobial resistance in bacterial chronic sinusitis. AB - Recent reports describe the emergence of antimicrobial resistant bacteria in acute sinusitis and an increased incidence of enteric gram negative bacilli in chronic sinusitis. The objective of this cross sectional study is to identify the emergent resistance patterns in bacterial chronic sinusitis. Specifically, this article seeks to characterize the bacteriology of outpatient chronic sinusitis, then to compare the antimicrobial susceptibilities of the bacterial isolates with standard culture data from a tertiary care center. Between March and August, 1994, 113 new outpatients presented with chronic sinusitis at a major teaching institution. Of these patients 34 underwent endoscopically guided aerobic culture of the paranasal sinuses and nasal cavities. Of the 48 total cultures, there were 43 positive cultures yielding 72 isolates. Thirty-eight cultures had two or fewer isolates; four cultures had three plus isolates, and one culture grew out normal flora. The most frequently isolated organisms were coagulase negative Staphylococcus (SCN), 20 (28%); Pseudomonas aeruginosa, 12 (17%); and Staphylococcus aureus, 9 (13%). Within the limited sample size for each isolate, Staphylococcus coagulase negative, Pseudomonas, and Pneumococcus demonstrated higher antimicrobial resistance compared to the medical center's corresponding nonurinary isolates. Additionally, three of six patients with Pseudomonal aeruginosa (50%) had a quinolone resistant strain. These preliminary data suggest that both an increased incidence of antimicrobial resistance and of enteric gram negative bacilli may exist in these outpatient, tertiary care center patients with chronic bacterial sinusitis. PMID- 9740917 TI - Functional endoscopic sinus surgery: do ratings of appropriateness predict patient outcomes? AB - Sinus surgery appropriateness ratings were recently developed by Value Health Sciences in cooperation with AAO-HNS. The goal of this study was to assess the relationships among three ratings of sinus surgery appropriateness (Appropriate, Inappropriate, or Equivocal) and symptom response. The enrolled population included 49 patients who completed the Smo-Nasal Outcome Test-20 (SNOT-20) presurgery and 6 months postsurgery. The SNOT-20 is a patient-based measure of sinusitis-related health status and quality of life. Overall, the mean percent difference (delta %) between pre- and postsurgery SNOT scores was 38%, a statistically and clinically significant improvement. However, there was no relationship between the appropriateness rating for the surgery and the delta % SNOT score (i = 1.83, p-value = 0.171); 20 patients with an Equivocal rating demonstrated the greatest delta % (49%), 20 patients with an Appropriate rating showed the least delta % (26%), and 9 patients with an Inappropriate rating had an intermediate delta % (39%). Furthermore, those patients having an Appropriate rating at the time of surgery reported a greater persistence of bothersome symptoms at 6 months (p-value = 0.02) then patients in either the Equivocal or Inappropriate rating. These results suggest that appropriateness ratings may not predict which patients will obtain the greatest symptom improvement from sinus surgery. PMID- 9740918 TI - Chronic sinusitis: a sequela of inferior turbinectomy. AB - Inferior turbinectomy has generated a great deal of controversy among rhinologic surgeons. Proponents of partial and total inferior turbinectomy cite numerous studies of large numbers of patients with subjective relief of nasal obstruction after turbinectomy. Clinical studies critical of turbinectomy have focused on complications such as hemorrhage, crusting, adhesions, and atrophic rhinitis. Our study was undertaken to evaluate the incidence of chronic sinusitis post inferior turbinectomy. Postoperative evaluation by history, physical examination, and computerized tomography of the paranasal sinuses revealed that a significant number of patients who underwent inferior turbinectomy developed sinusitis. Patients evaluated in our clinic for nasal obstruction underwent a detailed history, physical examination along with nasal endoscopy and coronal computerized tomography of the paranasal sinuses. Those patients with nasal obstruction not responsive to medical treatment and without evidence of sinusitis underwent submucous resection and inferior turbinectomy. The incidence, cause, and possible prevention of post inferior turbinectomy sinusitis is discussed in this article. PMID- 9740919 TI - Comprehensive management of allergic fungal sinusitis. AB - In little more than a decade, allergic fungal sinusitis has gone from a medical curiosity to one of the more perplexing problems to challenge the otorhinolaryngologist. These patients are typically immunocompetent adolescents or young adults with pansinusitis (unilateral and bilateral) and polyposis, atopy, and characteristic radiographic findings. Allergic mucin contained within the sinuses demonstrates numerous eosinophils and Charcot-Leyden crystals, and fungal stains show the presence of noninvasive hyphae. Fungal cultures may or may not be positive. We have found the following approach to allergic fungal sinusitis to be most effective: 1) Adequate preoperative evaluation and medical preparation; 2) Meticulous exenterative surgery; 3) Closely supervised immunotherapy with relevant fungal and non-fungal antigens; 4) Medical management including topical and systemic corticosteroids as needed; 5) Irrigation and self cleansing by the patient; and 6) Close clinical follow-up with endoscopically guided debridement when necessary. PMID- 9740920 TI - A study of the association between epistaxis and the severity of hypertension. AB - Hypertension (HTN) has frequently been cited as a general risk factor for epistaxis. However, studies dealing with this association have yielded equivocal results. In this study, a sample of 121 hypertensives (blood pressure > or = 140/90 mmHg) was selected to evaluate the association between the severity of HTN and a previous history of epistaxis. Patients with an average blood pressure > or = 160/100 mmHg were classified as suffering from a more severe form of HTN and were compared with those with a less severe form of the disease (160/100 mm Hg < or = blood pressure > or = 140/90 mm Hg). The frequency of epistaxis did not differ among patients categorized by the severity of HTN. Users of aspirin were found to be twice as likely to have a history of epistaxis. In addition, there was a statistical tendency for an association between a history of epistaxis and the duration of hypertension. We conclude that the severity of HTN and a history of epistaxis were not associated in a cohort of hypertensive patients. The identification of other risk factors for epistaxis, including the duration of HTN, deserves further study. PMID- 9740921 TI - Epiphora: the role of rhinitis. AB - Inflammation in the nasal mucosa may lead to epiphora by causing edema around the orifice of the nasolacrimal duct. We present and discuss three cases where simple treatment of rhinitis led to the resolution of the presenting symptom of epiphora, avoiding the need for surgery. A randomized prospective trial of topical nasal corticosteroids in the management of epiphora associated with rhinitis is in progress. PMID- 9740922 TI - Nonspecific nasal mucosal reactivity, expressed as changes in nasal airway resistance after bilateral saline provocation. AB - When using intranasal provocation tests in diagnosing nasal allergy or other hyperreactivity, it is essential to know which part of the reaction is caused by some nonspecific stimulus. The purpose of this study is to evaluate nonspecific nasal reactions of patients with chronic nonallergic perennial rhinitis as well as normal controls. The provocations were made bilaterally with normal saline solution (0.9% sodium chloride), and the changes in the nasal resistance were recorded by active anterior rhinomanometry. There was no significant difference between the chronic rhinitis patients and the normal controls. However, the individual variations were large. We conclude that a change of +/- 100-150% in the unilateral nasal resistance can be caused by a nonspecific reactivity. PMID- 9740923 TI - Effects of IL-1 beta, TNF-alpha, and TGF-beta on proliferation of human nasal epithelial cells in vitro. AB - Previous reports suggest that cytokines may be involved in proliferation of the epithelium. The aim of this study was to determine the effects of cytokines, IL-1 beta, TNF-alpha, and TGF-beta on proliferation of human nasal epithelial cells (HNECs) in vitro. Primary cells were cultured from HNECs on collagen gel matrix. Subcultured HNECs were incubated in a medium with recombinant human (rh) cytokines, rhIL-1 beta, rhTNF-alpha, and rhTGF-beta at different concentrations of 0.01 ng/mL, 0.1 ng/mL, 1 ng/mL, 10 ng/mL, and 100 ng/mL. After 2-day incubation with these cytokines, daily cell proliferation was measured by MTT assay for 6 days. While rhIL-1 beta inhibited proliferation of HNECs in concentration-dependent and time-dependent manners, rhTNF-a stimulated HNEC growth at concentrations ranging from 0.01 ng/mL to 10 ng/mL in concentration dependent and time-dependent manner. In contrast, rhTGF-b inhibited HNEC growth irrespective of concentration and incubation time. This study suggests that IL-1 beta, TNF-alpha, and TGF-beta may have an important role in the repair of the nasal mucosa by regulating proliferation of the nasal epithelium. PMID- 9740924 TI - Transnasal endoscopic approach to the sella turcica. AB - The transseptal/transsphenoidal approach to the pituitary gland has been the most commonly used approach for resection of pituitary adenomas for the last 50 years. This procedure has a low morbidity and provides direct midline access to the sella and pituitary gland. Recent advancements in endoscopic surgery, however, suggest that a lower morbidity approach to the sella would be possible via transnasal endoscopic route. Prior reports have confirmed effectiveness of this approach to the pituitary gland and we report here an early series of endoscopic transnasal pituitary surgery from our institution. We report seven cases of transnasal endoscopic pituitary surgery. Our technique consists of endoscopic exposure of the sphenoid ostium unilaterally, excision of the posterior septum anterior to the rostrum of the sphenoid sinus with resection of the sphenoid rostrum for bilateral exposure of the sphenoid sinus. A specially designed nasal speculum is positioned to displace the posterior septum and lateralize the middle turbinates, permitting direct midline exposure of the sphenoid sinus and sella. We have progressively modified the technique over the seven cases that we present and will discuss our specific instrumentation, indications, and technique for this procedure. We have encountered one cerebrospinal fluid leak in this series. Patient satisfaction has been high and hospitalization is less than with the conventional transseptal approach, averaging 1 day. Our impression is that the transnasal endoscopic approach to pituitary adenomas is a safe technique with reduced morbidity permitting shortened hospital stay. PMID- 9740925 TI - Rhinocerebral mucormycosis caused by Apophysomyces elegans. AB - Mucormycosis is an uncommon fungal disease and one of the most fulminant infections known. This is the second report of rhinocerebral mucormycosis caused by Apophysomyces elegans, a newly recognized genus and species classified in the family Mucoraceae. The patient was a 54-year-old man being treated for a severe sinus infection with antibiotics and oral steroids. Recovery occurred in our patient after prompt surgical debridement and drainage of his maxillary sinuses. This case fits the reported characteristics of other A. elegans infections including warm climate, intimate contact with the soil, and an incubation period measured in days. Several reported cases indicate A. elegans can cause mucormycosis in immunocompetent individuals with no underlying medical problems. PMID- 9740926 TI - Brompheniramine maleate: a double-blind, placebo-controlled comparison with terfenadine for symptoms of allergic rhinitis. AB - This was a double-blind, randomized, placebo-controlled, multicenter, parallel study comparing the effectiveness, at recommended doses, of an extended-release formulation of brompheniramine maleate and terfenadine in the treatment of allergic rhinitis. Subjects with symptoms of seasonal and/or perennial allergic rhinitis received brompheniramine 12 mg (n = 106), 8 mg (n = 105), terfenadine 60 mg (n = 106), or placebo (n = 53) twice daily for 14 days. On treatment days 3, 7, and 14, symptom severity ratings (i.e., rhinorrhea, sneezing, nasal congestion, itchy nose, eyes or throat, excessive tearing, postnasal drip) were completed by the physician; subjects and physicians each completed a global efficacy evaluation. Brompheniramine 12 mg and 8 mg and terfenadine were more effective than placebo (p < or = 0.05) on the physicians' global: brompheniramine 12 mg was more effective than terfenadine (p < or = 0.05) on days 7 and 14 and brompheniramine 8 mg on day 3. On the subjects' global evaluation, brompheniramine 12 mg and 8 mg and terfenadine were more effective than placebo (p < or = 0.05); brompheniramine 12 mg was more effective than terfenadine (p < or = 0.05) on days 7 and 14 and brompheniramine 8 mg on day 3. In general, brompheniramine 8 mg was comparable to terfenadine. On days 3 and 7, the total symptom and total nasal symptom severity scores for subjects receiving brompheniramine 12 mg were significantly more improved than for placebo (p < 0.05); terfenadine was not different from placebo; brompheniramine 12 mg was significantly better than terfenadine on day 7 (p < 0.05) for reducing total symptom severity and on days 3, 7, and 14 for reducing total nasal symptom severity. Adverse experiences were reported by 155 (41.9%) of the 370 subjects enrolled in the study. The overall rate of adverse experiences in the brompheniramine 12 mg treatment group (57.5%) was significantly greater (p < 0.05) than for brompheniramine 8 mg (38.1%), terfenadine (31.1%), and placebo (39.6%). In conclusion, an extended-release formulation of brompheniramine 12 mg or 8 mg bid alleviates allergic rhinitis symptoms and brompheniramine 12 mg provides significantly better relief of these symptoms than terfenadine 60 mg bid. PMID- 9740927 TI - [Cytokinins, helper cells and killer cells. Reduced immune function by anesthetics]. PMID- 9740928 TI - [Anesthesia and perioperative immune function]. AB - Innate and acquired immunity plays a pivotal role in the host defense response. Pain, stress, necrotic tissue and invading microorganisms are known modulators of the complex immune response of patients undergoing major surgery. Anaesthesia itself or perioperative interventions of the anaesthesiologist may substantially alter the immune function with potential impact on the postoperative course. For instance, transfusion of allogenic blood and administration of dopamine or metoclopramide may interfere with immunity. Stress and pain are associated with immune tolerance, increased susceptibility to infection and tumor spreading in animal models. Thus, anaesthesia may--through modulation of the neurohumoral stress response--indirectly affect immunity of the surgical patient. In particular epidural anaesthesia and/or administration of epidural or spinal opioids seem to attenuate the stress response with beneficial effects on cellular and humoral immunity. In addition, anaesthetics, such as etomidate, propofol, or thiopentone and opioid analgesics may directly affect function of immune competent cells. However, these actions may only be apparent with high or supraclinical concentrations and/or long-term exposure. Regarding the latter, evidence suggests that long-term sedation using thiopentone in neurosurgical patients is paralleled by infectious complications in a dose-dependent manner. At present, no data are available regarding the significance of the observed alterations associated with various anaesthetic procedures of the incidence of postoperative complications associated with impaired immunity, such as infection or metastatic spreading in oncological surgery. PMID- 9740929 TI - [Anesthesia for cesarean section in Germany. A survey]. AB - AIM OF STUDY: Goal of this survey is to give an overview of anaesthesia for caesarean section in Germany. METHOD: In 1994 and 1995, we sent a questionnaire to the chief-anaesthetists of all German hospitals with departments of gynaecology/obstetrics to find out the routine anaesthetic procedures for caesarean section. RESULTS: We obtained data from 409 hospitals (response rate 46.4%) with 321,816 births--50,123 of which were sections (mean caesarean section rate 16.6%). The mean general anaesthesia rate for elective caesarean sections was 66.5%, for non-elective sections 90.8%. The mean epidural anaesthesia rate for caesarean section was 22.6% and the mean spinal anaesthesia rate was 9.8%. For general anaesthesia most hospitals used antacids and/or histamine2-receptor antagonists (64.6% of responding hospitals). Anaesthesia was induced with intravenous barbiturates (82%), succinylcholine for intubation (98.2%) and no opioids before clamping of the cord (94.8%). For regional anaesthesia bupivacaine was the most common local anaesthetic (spinal 84.0%, epidural 96.8%). Opioids were added to local anaesthetics for epidural anaesthesia at 21.4% of the hospitals. CONCLUSIONS: General anaesthesia is the commonest practice for caesarean sections at German hospitals. Nowadays regional anaesthesia gains more importance compared to previous German surveys and in agreement with foreign data. PMID- 9740930 TI - [Dose-response relationship of clonidine with epidural administration of ropivacaine in orthopedic procedures of the lower extremities]. AB - OBJECTIVE: The aim of this study was to investigate preliminarydose-range effects of clonidine added to ropivacaine for epidural analgesia in elective orthopedic surgery of the lower limbs with doses, causing a minimum of cardiovascular side effects. METHODS: 60 patients were randomly assigned to receive in a double-blind fashion a mixture of 1 mg/cm height ropivacaine plus saline or 1 mg/cm ropivacaine plus 25 micrograms, 50 micrograms, 75 micrograms, 100 micrograms or 150 micrograms clonidine for epidural analgesia. The sensory and motor function were determined at defined time intervals for 30 minutes. Heart rate and blood pressure were controlled and sedation score was judged. The postoperative 2 segment-regression of pin-prick and the onset of pain were recorded. RESULTS: The six groups were comparable in demographic data and in term of onset time. The prolongation of analgesia reached 513 +/- 92 min (p = 0.002) for 150 micrograms clonidine, 460 +/- 148 min (p = 0.073) for 100 micrograms clonidine, 440 +/- 86 min (p = 0.057) for 75 micrograms clonidine compared with 347 +/- 114 min for saline. In an equal manner, 2-segment-regression for pin-prick was extended to 251 +/- 47 min (p = 0.018) for 150 micrograms clonidine, 238 +/- 33 min (p = 0.034) for 100 micrograms clonidine, 229 +/- 29 min (p = 0.027) for 75 micrograms clonidine and 178 +/- 43 min for saline. Heart rate dropped down in all groups. Mean arterial pressure decreased significantly in the groups with 75, 100 and 150 micrograms clonidine. Sedation score increased continuously from 0.6 +/- 0.5 (saline) to 1.8 +/- 0.8 (150 micrograms clonidine). CONCLUSION: We conclude that 150 micrograms clonidine significantly enhances the duration of analgesia of epidurally administered ropivacaine in a mean of 171 mg. This time interval is longer than the one with 200 mg ropivacaine alone. But, there are side effects in form of decrease of arterial pressure. Cardiovascular monitoring seems to be essential. Because of the enhanced analgesia duration, the time interval for reloading epidural anaesthesia are increased. PMID- 9740931 TI - [Reduced muscular oxygen tension and nerve impulse transmission from antishock hose. Reduction of oxygen tension in the tibial muscle and impulse transmission in the peroneal nerve from pneumatic -1 pressure from antishock hose]. AB - OBJECT OF THE STUDY: The aim of the study was to assess, whether the pneumatic pressure of an antishock-trouser (AST) of 20-40 mm Hg induces a decreased oxygenation of the anterior tibial muscle and attenuates muscular response potential (MRP) of n. peronaeus profundus? METHODS: Among 22 normotensive, healthy volunteers the AST were tested by applying pressure values between 0 and 100 mm Hg and measuring the intracompartmental pressure, the muscular oxygen pressure as well as the MRP by electroneurographic means within a period of 6 hours. RESULTS: The median initial intracompartmental pressure value of the m. tibialis anterior was 12.0 mm Hg (Q25%/Q75%: 8.9/17.3), the muscular oxygen pressure 14.8 mm Hg (Q25%/Q75%: 11.5/22.0). Transmission of the pneumatic AST-leg segment pressure to the muscle: 97.7% (Q25%/Q75%: 89.2/99.8). Already in the low AST pressure field (20-40 mm Hg) a severe hypoxia occurred in one case. A reduction of MRP was noticed at an AST pressure rate of 10 mm Hg. In 5 of 6 cases AST pressure values of 60 mm Hg led to pathological pO2-values within 5-20 minutes. Almost without exception AST-pressure rates < 60 mm Hg resulted in an anoxia of the muscle and loss of the MRP. CONCLUSIONS: We should demand that the AST are only applied with models where the pressure generated within the single segments can be controlled by pressure gauge. The application of the AST seems to be justified for polytraumatised in severe haemorrhagic shock where the risk of a local tissue ischemia with systemical consequences must deliberately be accepted. PMID- 9740932 TI - [Procalcitonin. A new diagnostic parameter for severe infections and sepsis]. AB - Procalcitonin (PCT), a glycoprotein consisting of 116 amino acids, has been proposed as a new marker of severe infection. The site of production under this condition remains unknown. The serum PCT concentration is determined by an immunoluminometric assay of 40 microliters serum or plasma requiring approximately two hours. Elevations of PCT are for instance associated with levels of lipopolysaccharide and the cytokines TNF-alpha and IL-6. Bacterial, parasitic or fungal infections developing septic complications in contrast to local infections, often show values exceeding 2 ng/ml. The specificity of the parameter in this context increases with its concentrations. Therapeutic actions that confine the infection locally are reflected by a decrease of the PCT value. PCT may be elevated within the first days after extended surgery or polytrauma, in some malignancies, heat-stroke and during treatment of some hematologic diseases without an existing sepsis or severe infection. Previous studies indicate certain benefits of PCT compared to traditional markers of inflammation or sepsis, where the ability to indicate a generalized infection is the primary advantage. PMID- 9740933 TI - [Psychiatric emergencies from the viewpoint of the emergency physician]. AB - OBJECTIVE: In the German physician-based emergency medical system (EMS) psychiatric emergency situations (PES) rank on third place contradictory to it's importance during emergency physician training program. The aim of our study was to examine the relevance of PES and the stress which PES imposes upon EMS physicians. Further, the interest of training programs on that issue was determined. Knowledge about PES was investigated by a short test. METHODS: 952 emergency physicians were sent a questionnaire about following: demographic data, frequency of PES, strain by PES, own knowledge, interest about training programs. Further five typical PES were presented for diagnostic and therapeutic judgement. RESULTS: 222 responded (183 men/37 women/2 without gender data, average age: 40.1 +/- 6.7, qualification as emergency physician: 9.6 +/- 5.1 years, most frequent subspeciality in-hospital physicians: anaesthesiology 67.5%, in-practice physicians: general medicine 72.1%). PES frequence was estimated at 9.4%, personal knowledge judged only by 13% as sufficient, 14.2 felt incapable by PES. 73% saw importance of training, especially expressed by the more experienced (P < 0.05). Test presented 65% correct diagnoses, 33% correct therapy, 26% incorrect decision of hospital admission. CONCLUSION: PES are a frequent problem of pre hospital patient care for emergency physicians. As personal knowledge was estimated to be insufficient, the interest for courses concerning PES issues is high. PMID- 9740934 TI - [Vertical infraclavicular brachial-plexus blockade. A clinical study of reliability of a new method for plexus anesthesia of the upper extremity]. AB - We examined the efficacy of the vertical infraclavicular block for plexus brachialis anaesthesia using a nerve stimulator after introducing the method (VIP1) and after three years of clinical experience (VIP2). In two prospective studies we compared the results with each other as well as with the efficacy of the axillary block (AX). At VIP1, we found a complete analgesia in 88% of the patients, whereas in 9% a supplementation was needed. In group AX the results were significantly worse (complete: 70%, supplementation: 24%; p < 0.001). No increase of the rate of efficacy could be found when having some clinical experience with the VIP (VIP2: complete 87%, supplement: 11%). In general, the results of the VIP depended on the motoric answer to the nerve stimulation. There were no complications of the VIP such as nerve lesions or pneumothorax. The VIP using a nerve stimulator is a simple, reliable and uncomplicated method for plexus-brachialis-anaesthesia, which is easy to learn. PMID- 9740935 TI - Temperature of the cerebrovenous blood in a model of increased intracranial pressure. AB - Hypothermia has a considerable protective effect during brain ischemia. On the other hand small increases of brain temperature have a remarkable effect on the exacerbation of neurological damage following an ischemic event. Hyperthermia of the brain tissue after severe head injury is described. The effect of acutely increased intracranial pressure on cerebrovenous blood temperature is not described yet. The aim of this study was to investigate the relationship between temperature in the cerebrovenous compartment (Tcv) and changes of the CPP in an animal model of raised intracranial pressure. METHODS: A thermocouple was inserted in the sagittal sinus in 9 pigs under general anesthesia. By stepwise inflating a supracerebral and infratentorial placed balloon catheter intracranial pressure (ICP) was increased and CPP concomitantly decreased. The central body temperature was measured simultaneously in the abdominal aorta (Ta) with a second thermocouple. RESULTS: In our model th Tcv was lower than Ta at the beginning of the ICP increase. The mean difference between Ta and Tcv, (delta Ta-cv) was 0.86 degree C (+/- 0.44) prior to ICP increase and 1.19 degrees C (0.58) at the maximum ICP increase. Thus, delta Tav increased during CPP reduction. This relation was represented by an adjusted R(square) of r2 = 0.89 (p < 0.001). CONCLUSIONS: The CPP decrease, caused by an increasing ICP, results in changes of the cerebrovenous blood temperature. Interpreting the present results the experimental situation of a relative colder cerebral compartment in comparison to the central body temperature has to be considered. However, the results imply, that simultaneous temperature monitoring of the central body temperature and the cerebrovenous blood temperature is an additional source of information about relative changes of the CBF. PMID- 9740936 TI - [Epidural fentanyl: legal, illegal, does it matter? Remarks on the work of J. Jage and H. Hartje. Postoperative pain therapy, Part II. Anaesthesist (1997) 46:161-173]. PMID- 9740937 TI - [Reader's questions--expert answers]. PMID- 9740938 TI - [Weaning from artificial respiration. 1]. PMID- 9740939 TI - Elements of a trusting relationship. PMID- 9740940 TI - A pilot study of esthetic perceptions of dental fluorosis vs. selected other dental conditions. AB - The prevalence of fluorosis has increased over the past fifty years, and with this increase, esthetic concerns pertaining to fluorosis should also be taken into consideration. Canadian, Australian, and British studies have explored perceptions concerning enamel fluorosis, but no studies in this area have been published from the United States. In the previous studies, esthetic concerns resulting from fluorosis generally were not compared with the esthetic perceptions of other conditions such as isolated opacities, tetracycline staining, or various types of malocclusion. In the present investigation, respondents answered written questions about paired photographs, one of fluorotic teeth and the other with one of the other conditions. Results show that not only is fluorosis noticeable, but it may be more of an esthetic concern than the other conditions. PMID- 9740941 TI - Comparative in vitro microradiographic effects of resin-modified and autopolymerizing glass ionomers on demineralization of primary and permanent enamel. AB - The purpose of this study was to compare in vitro effects of resin-modified and autopolymerizing glass ionomer restorative materials on demineralization of primary and permanent human enamel. Thirty primary and permanent enamel specimens measuring approximately 3 x 4 x 4 mm were sectioned and plano-paralleled before random placement of materials: Photac-Fil, a resin-modified glass ionomer; Ketac Fil, an autopolymerizing glass ionomer; and Tytin, a silver amalgam. After incubation for twenty-four hours, the samples were pH cycled for eight hours at pH 5.0 and sixteen hours at pH 7.2 for a total of two weeks, all at 37 degrees C. The specimens were then subjected to an artificial caries challenge at pH 5.0 for 196 hours. The specimens were embeded in Epon 812 and incubated at 55 degrees C for thirty-six hours. Microsections were produced from each sample and subjected to microradiography and quantitative microdensitometry. Data on lesion depth and mineral content were analyzed by Two Way ANOVA and Student Newman-Keuls Pairwise Multiple Comparison tests. There were significant differences in lesion depth and mineral content between groups (p < 0.05) and between permanent and primary enamel. This study demonstrates that Photac-Fil and Ketac-Fil prevent in vitro demineralization at varying levels in primary and permanent enamel. PMID- 9740942 TI - Antibiotic prophylaxis in dental patients with ventriculo-peritoneal shunts: a pilot study. AB - Fourteen hydrocephalic children with ventriculo-peritoneal shunts received routine dental prophylaxis and topical fluoride application. No antibiotics were administered to these children for any reason during the three months before treatment or during the twelve months after treatment. None of these children presented with any signs of shunt infection during the twelve-month posttreatment period. In spite of the small sample size, this prospective pilot study suggests that patients with ventriculo-peritoneal shunts are not susceptible to shunt infection following a bacteremia induced by a dental prophylaxis and topical fluoride treatment. Dental prophylaxis without antibiotic coverage in patients with V-P shunts, therefore, does appear safe. We recommend that further study with a larger population, or a collaborative study by several medical centers, be performed to establish more conclusively that prophylactic antibiotics are not necessary for patients with ventriculo-peritoneal shunts who receive dental procedures. In addition, other investigations are needed to determine the risk of shunt infection with more invasive dental procedures, such as periodontal surgery or tooth extraction. PMID- 9740943 TI - Assessment of the caries activity test (Cariostat) based on the infection levels of mutans streptococci and lactobacilli in 2- to 13-year-old children's dental plaque. AB - It is generally agreed that mutans streptococci and lactobacilli are associated etiologically with dental caries. The caries activity test, Cariostat, was designed to measure the pH decrease caused by microorganisms in the plaque sample obtained from the buccal surfaces. Researchers found the test to be a reliable, diagnostic, and predictive device. Incubation was done on MS and MSB plates in an atmosphere of 95 percent N and 5 percent CO at 37 degrees C and for 48 hours. The relationship of the Cariostat scores and the pH values are shown in a table. The test scores are shown for two age-groups: Ages two-to-six years with primary dentitions, and ages five-to-thirteen years with mixed dentitions. The advantages of the Cariostat test are: the sampling method is simple and the time of analysis is short; the test can be used for the very young and for patients difficult to manage; and it requires no specialized knowledge or equipment. PMID- 9740944 TI - Self-report measurements of dental anxiety and fear in children: a critical assessment. AB - This article reviews self-report measurements frequently used to assess dental anxiety in children. The main focus is on their reliability and validity. For this purpose correlations between the reviewed measurements and other measurements of dental fear in children are considered, as well as their possible ambiguity with respect to scoring procedures and their ability to discriminate between fearful and non-fearful children. Results show that all three questionnaires discussed are open to criticism. It is concluded that of the self report measurements, the Children's Fear Survey Schedule-Dental Subscale (CFSS DS) is to be preferred to both Corah's Dental Anxiety Scale (DAS) and the Venham Picture Test (VPT). The reasons for this are the following: the CFSS-DS covers more aspects of the dental situation; it measures dental fear more precisely than the other scales; normative data are available on this scale; and it has slightly superior psychometric properties. PMID- 9740945 TI - How's your Spanish, Vietnamese, Hmong and ...? Or, can your pediatric patients understand you? AB - While there are areas of concentration in particular states, throughout the nation the number of young children (and their parents/guardians) who have limited communication abilities in the English language is continuing to increase. Almost three-quarters of school-age children with limited English language proficiency speak Spanish at home. PMID- 9740946 TI - Are we maintaining the ratio of private practicing pediatric dentists to the number of children? AB - Using recent ADA and Bureau of the Census reports, national, region and state ratios of pediatric dentists per 100,000 children are provided for the mid-1990s. While national averages continue to improve, there are wide variations at more local levels. PMID- 9740947 TI - Bilateral twinning: report of case. AB - Twinning is an anomaly in which one tooth has combined with another or enlarged itself to the point of doubling its substance. A discussion of the nomenclature and a report of bilateral twinning are presented. A possible cause of the twinning events in this case include gemination of the maxillary right and mandibular left canines and fusion of the maxillary left and mandibular right canines with supernumerary teeth. PMID- 9740948 TI - Infraclusion of lower primary molar with other familial dental anomalies: report of case. AB - Impaction of a primary tooth is a rare occurrence compared to impaction of a permanent tooth. Impaction should be differentiated from reimpaction, a condition in which an erupted tooth loses its apical movement and impaired development of the alveolar bone surrounding the tooth occurs. In dental ankylosis, the height of the alveolar process stops in the affected area, the adjoining teeth continue to move occlusally, and neighboring teeth drift. The author discusses the uncertain etiology of reimpaction. The history of a three-year-old patient is presented. PMID- 9740949 TI - An introspective qualitative report on dietary patterns and elevated levels of dental decay in a deprived urban population in northern Mexico. AB - Disorganized urbanization in Latin America has led to masses of impoverished people to become squatters in the larger urban areas. Using a community development network in the outskirts of Tijuana, in Northern Mexico, this investigation assessed the dental health situation, aiming to establish the underlying behavioral causes of poor oral health in these slums. Using quantitative and qualitative tools, fifty-six mothers (mean age 30.1 +/- 7.2) with their accompanying children (n = 56; mean age 6.1 +/- 3.3; 46.4 percent female) were interviewed and examined. Dental health was poor and characterized by vast unmet treatment needs in adults and children. 22.2 percent of children under three years of age suffered from Early Childhood Caries, strongly linked to inappropriate patterns of bottle use. Dietary patterns for the overall child population included many cariogenic snacks and beverages. A straightforward model to explain behavioral structures incorporates these findings against the background of living in a highly-deprived environment, whereby the allure of more affordable gratifications for self and family is often translated in the form of tokens such as junk food. PMID- 9740950 TI - The other Calgary declarations. PMID- 9740951 TI - Update in glaucoma: the new pharmacotherapies. Dorzolamide hydrochloride. PMID- 9740952 TI - Update in glaucoma: the new pharmacotherapies. Brimonidine versus apraclonidine. PMID- 9740953 TI - Update in glaucoma: the new pharmacotherapies. Latanoprost. PMID- 9740954 TI - Catecholaminergic nerve fibres in normal and alkali-burned rabbit cornea. AB - BACKGROUND: In recent years anatomic research has demonstrated the presence of various types of nerve fibre in the cornea. The purpose of this study was to investigate the distribution of catecholaminergic fibres in various corneal layers and to study the effects of an experimental superficial corneal lesion on the pattern of catecholaminergic nerve fibre distribution in the various corneal layers. METHODS: Three weeks after the creation of an alkali burn in the centre of the right cornea of five albino rabbits, the animals were killed, and histologic sections from the cornea of both eyes were stained for observation of catecholaminergic nerve fibres and photographed on black-and-white film. The photographs were examined using the Quantimet image analyser (Leica). RESULTS: Catecholaminergic nerve fibres were observed in the corneal epithelium and the deep stromal layers. Sections from injured cornea showed a drastic reduction in epithelial and superficial stromal catecholaminergic nerve fibres, whereas the nerve fibres in the endothelium and deep stroma were not damaged. INTERPRETATION: Although catecholaminergic nerve fibres were observed in all corneal layers, the pattern of catecholaminergic fibres following the creation of a superficial lesion of the cornea seems to suggest that the superficial and deep nerve fibres may have a different distribution. PMID- 9740955 TI - Effect of cataract surgery with intraocular lens implantation on inflammation in chronic uveitis: a longitudinal laser flare photometry study. AB - BACKGROUND: The precise effect of cataract surgery with implantation of an intraocular lens (IOL) on the course of uveitis is not well known. Laser flare photometry allows quantitative assessment of intraocular inflammation. The aim of this study was to determine the effect of cataract surgery with IOL implantation on the disease course and level of inflammation in chronic uveitis, using laser flare photometry monitoring. METHODS: The charts of all patients who underwent surgery for secondary uveitic cataract between 1990 and 1994 (24/558 [4.3%]) were reviewed. Only eyes that had received standardized perioperative steroid treatment and had systematic laser flare photometry follow-up were included. Cataracts due to Fuchs' heterochromic cyclitis were excluded. Visual acuity, flare values and recurrence of flare episodes were compared before and after cataract surgery, and postoperative data were compared between eyes that received heparin-coated IOLs and those that received uncoated IOLs. RESULTS: Nineteen eyes of 16 patients met the inclusion criteria. The mean length of the pre- and- postoperative follow-up periods was 779 and 444 days respectively. The mean visual acuity increased from 0.2 (standard error of the mean [SEM] 0.2) preoperatively to 0.8 (SEM 0.3) postoperatively (p < or = 0.001). The mean flare value decreased from 58.6 (SEM 18.6) photons/ms during preoperative follow-up to 29.7 (SEM 7.8) photons/ms during postoperative follow-up (p < or = 0.006). The mean number of recurrences per 6 months decreased from 0.27 (SEM 0.03) preoperatively to 0.12 (SEM 0.01) postoperatively (p < or = 0.05). The difference in the postoperative recurrence rate between the eyes that received coated IOLs (0.0) and those that received uncoated IOLs (0.18 [SEM 0.02]) approached statistical significance (p < or = 0.054). INTERPRETATION: Quantitative assessment of inflammation by laser flare photometry in patients undergoing surgery for uveitic cataract showed that there was significantly less inflammation and fewer recurrences postoperatively and that recurrences were less severe. PMID- 9740956 TI - Neuro-ophthalmic findings in progressive multifocal leukoencephalopathy. AB - BACKGROUND: Progressive multifocal leukoencephalopathy (PML) is a demyelinating disorder of the central nervous system found in immunodeficient patients, most frequently now in those infected with HIV. It may represent the initial manifestation of HIV infection. Since the central visual pathways may be affected, a variety of neuro-ophthalmic signs and symptoms can manifest. We studied the clinical, radiographic and histopathological characteristics of patients with PML. METHODS: The charts of 13 patients in whom PML was diagnosed in the Neuro-AIDS clinic at the Montreal Neurological Institute between November 1987 and March 1995 were reviewed. The diagnosis of PML was established by characteristic clinical features together with typical computed tomographic or magnetic resonance imaging findings, such as nonenhancing low-density (on computed tomography) or hyperintense (on T2-weighted magnetic resonance imaging) white-matter lesions, without mass effect. Neuro-ophthalmic findings were based on clinical examination by an ophthalmologist, neuro-ophthalmologist or neurologist. Tissue for pathological examination was obtained by biopsy in one case and at postmortem study in a second case. RESULTS: The most common finding was homonymous hemianopia, in five patients (38%). Other features included nystagmus (in two patients), diplopia with cranial nerve palsy (in one) and cortical blindness (in one). One of the patients exhibited involvement of the brain stem, a site not usually affected by this demyelinating process. INTERPRETATION: The diagnosis of PML should be considered in immunocompromised patients with neuro-ophthalmic findings, particularly those with homonymous hemianopia. PMID- 9740957 TI - Prostate adenocarcinoma presenting as a solitary choroidal metastasis. PMID- 9740958 TI - Angle-closure glaucoma secondary to ciliochoroidal detachment. PMID- 9740959 TI - Scleritis associated with vitamin B12 deficiency. PMID- 9740960 TI - Role of the ophthalmologist as a patient educator. PMID- 9740961 TI - An epidemiologic investigation of unexpected refractive errors following cataract surgery. PMID- 9740962 TI - Reference materials: general aspects and IFCC strategy. International Federation of Clinical Chemistry. PMID- 9740963 TI - Criteria for the certification of internationally acceptable reference materials. PMID- 9740964 TI - Secondary reference materials. PMID- 9740965 TI - Selection of protocols for value assignment. PMID- 9740966 TI - Impact of reference materials on accuracy in clinical chemistry. AB - The analytical accuracy of the results of routine clinical chemistry measurements is contributed by a two-steps mechanism, involving transferring trueness from a higher metrological and monitoring the time-stability of trueness itself. In both operations, different materials are used: however, accuracy in the routine assay of genuine patient samples has to be the end product of this overall process. To such an aim, the materials must show an intermethod behavior similar to that of patient sera, i.e., they have to show commutability. Definitions of commutability and methods for assessing such a property are mentioned. The following aspects of lack of commutability of materials are then discussed: frequency; effects on the measured interlaboratory variability; and effects on the recalibration of analytical systems. The causes giving rise to lack of commutability are neither clear or easy to be shown. Matrix effect is one of the main causes; also, differences in the characteristics of the component being measured are often responsible for noncommutability of materials for enzyme activity measurements. Examples of these two different situations are given. It is concluded that, for an efficacious overall quality assurance process, either a set of minimally processed patient sera or commutable reference materials are to be used in the operations concerned with the control of trueness. An additional alternative approach is based on the use of materials with system-specific assigned values. PMID- 9740967 TI - BCR/IFCC reference material for plasma proteins (CRM 470). Community Bureau of Reference. International Federation of Clinical Chemistry. PMID- 9740968 TI - First International Reference Preparation for Individual Proteins in Urine. IFCC Working Group on Urine Proteins. International Federation of Clinical Chemistry. PMID- 9740969 TI - Reference material for PSA: the IFCC standardization study. International Federation of Clinical Chemistry. PMID- 9740970 TI - A reference system for cortisol. AB - OBJECTIVES: The International Federation of Clinical Chemistry (IFCC) initiated a pilot study, with cortisol as an example, that aims to implement the concept of standardization of hapten immunoprocedures on the basis of metrological traceability. In fact, this standardization concept comes down to correct calibration of measurement procedures, so that measurement results for patient samples can be traced back to the metrologically highest reference of a result, i.e., an SI unit as embodied in the primary reference material. In consequence, demonstration of standardization of a test system on the basis of traceability requires evaluation of measurement results pertaining to patient samples for accuracy. Such an evaluation shall be done by a correlation study of the routine test system with an accuracy-based reference measurement procedure. DESIGN AND METHODS: The IFCC project will select a panel of single donation human serum samples, and assign them with values for cortisol by measurement with at least two isotope dilution-gas chromatography/mass spectrometry reference methods. Limited amounts of the panel will be distributed to manufacturers, with the object to measure the samples with their calibrated immunoprocedures. The patient correlation studies between the routine and the reference methods will then be interpreted in terms of specificity and accuracy. It will also be investigated whether the performed comparison can be used as a basis for re-calibration. From the experience gained through this pilot study for cortisol, IFCC will formulate recommendations and guidance to manufacturers on how to perform and reliably interpret patient correlation studies. In this way, it is to be expected that the project might form a basis for general implementation of the concept of standardization of hapten immunoassays on the basis of metrological traceability. PMID- 9740971 TI - Interassay calibration as a major contribution to the comparability of results in clinical enzymology. AB - OBJECTIVE: Factors contributing to the applicability of interassay calibration of methods measuring enzyme catalytic activities are described. Also discussed are the properties essential for such a material. Similarity of specificity for the methods to be calibrated as well as commutability between the material(s) intended to be used as calibrator are the main criteria to be satisfied. RESULT: Several examples demonstrated that interassay calibration is feasible but a multi enzyme calibrator with a wide commutability for the most popular methods remains to be developed. This is the project of the IFCC Working Group on Calibrators in Clinical Enzymology (WG-CCE). Several experimental data are also presented that indicate that the temperature at which the reaction is carried out is not a limiting factor in the implementation of interassay calibration in clinical enzymology. PMID- 9740972 TI - Validation of an enzyme calibrator--an IFCC guideline. International Federation of Clinical Chemistry. AB - OBJECTIVES: The objective of this guideline is to improve standardization in clinical enzymology in order to improve intermethod comparability of patients' results. DESIGN AND METHODS: The reference system, combination of the reference method and the reference material, is used to produce a reference value for a given catalytic activity. Sets of methods are formed of methods exhibiting the same analytical specificity. Materials intended to be used as enzyme calibrators are experimentally checked for their commutability. RESULTS: The transfer of accuracy from the reference value to patients' results is dependent on methods (analytical specificity) and on materials (experimentally assessed commutability). The feasibility of this approach was demonstrated with materials of high level for several enzymes and for each of them for several routine methods. CONCLUSION: Expected advantages of this approach in clinical enzymology are presented. PMID- 9740973 TI - Prostate-specific antigen utilization in Ontario: extent of testing in patients with and without cancer. AB - OBJECTIVES: To ascertain the extent of prostate-specific antigen (PSA) testing in patients with prostate cancer (PC), with other cancers (OC), and with no cancer (NC) in two clinical laboratory databases. DESIGN AND METHODS: PSA test records were obtained from a tertiary care hospital, Sunnybrook Health Science Centre (SHSC) and from a private laboratory, Gamma-Dynacare Medical Laboratories (GDL), during the period 1988 to 1995. These records were linked with the Ontario Cancer Registry (OCR) to establish a diagnosis of PC, OC, or NC. Trends in PSA testing according to diagnostic category, testing laboratory, patient age (by decade), and PSA value (in microgram/L) were determined. RESULTS: Major cancer sites identified in the patients tested for PSA were prostate (60%), bladder and colon (7% each), lung (5%), kidney (3%), and rectum (3%). There were 11,867 patients (8.5%) with PC, 8,002 (5.9%) with OC, and 118,954 (86%) with NC. The total number of PSA tests performed on these patients was 230,756, of which 21% were on PC, 5% on OC, and 74% on NC; of these tests, 64% were performed through GDL and 36% through SHSC. The mean (median) number of tests per patient was: PC, 4.0 (2); OC, 1.4 (1); and NC, 1.5 (1). For PC 89% and for OC 72% of all tests occurred after diagnosis. Between 1990 and 1995 the number of PSA tests increased two-fold in PC and OC, and 20-fold in NC. We estimate that about one-half of the PSA tests in the NC group were for screening purposes. The proportion of PSA tests occurring in PC, OC, and NC for patients 50 to 70 years of age was 41%, 50%, and 63%, respectively; for patients over 70 years of age, this proportion was 58%, 46%, and 22% respectively; and for patients under 50 it was 1%, 4%, and 15%, respectively. Between 1990 and 1995, the largest increase in testing frequency was in the NC group, particularly in patients 50 to 70 years of age, which was accompanied by a decrease in patients over 70. Less than 10% of testing occurred in patients under 50 in all diagnostic groups. We estimate that about 26% of PSA screening tests in NC occurred outside the guidelines for patient age. Between 1988 and 1995, the proportion of PSA results below our detection limit (< 0.2 micrograms/L) showed a steady rise in the PC group, as did the proportion between 0.2 and 3.9 micrograms/L; these were accompanied by a fall in the proportion > 20.0 micrograms/L. However, the proportion of PSA results within these ranges did not change much during the same time period for the OC and NC groups. At cutoffs of PSA = 4.0 micrograms/L (or PSA = 10.0 micrograms/L), estimates of clinical specificity were 84.0% (or 96.3%), and of clinical sensitivity were 83.4% (or 47.1%). CONCLUSIONS: Most (86%) PSA testing occurred in men with NC, consistent with diagnosis or screening. There were more PSA tests per patient in PC than in OC, and most testing occurred after diagnosis. PSA testing in the NC group continues to increase rapidly. The proportion of PSA tests in patients over age 70 decreased in the order of PC > NC > OC. Between 1990 and 1995, there was an increase in the proportion of patients tested who were between 50 and 70 in the NC group, which may suggest more screening in this group. Over this same time period, there was an increase in the proportion of undetectable PSA values, possibly suggesting increased use of radical therapy; there was also a decrease in the proportion of PSA > 20 micrograms/L, possibly suggesting a decrease in the prevalence of advanced stage PC. PMID- 9740974 TI - Reference interval for human plasma nitric oxide end products. PMID- 9740975 TI - Depressive personality disorder: theoretical issues, clinical findings, and future research questions. AB - This article reviews the theoretical construct of depressive personality disorder and its related research. The history of depressive personality disorder is reviewed. It is concluded that differing theories converge on similar descriptions and mechanisms of development for the depressive personality disorder. Substantial empirical work supports the diagnostic distinctiveness of depressive personality disorder in clinical populations. Past and current assessment devices for assessing depressive personality disorder are also described along with their psychometric properties and clinical value. Suggestions are made for future research on the etiology and validity of the depressive personality disorder construct in order to facilitate deciding whether or not to include depressive personality disorder in future editions of the Diagnostic and Statistical Manual of Mental Disorders. PMID- 9740976 TI - Culture and classification: the cross-cultural application of the DSM-IV. AB - Changes incorporated into the latest edition of the Diagnostic and Statistical Manual of Mental Disorders (DSM-IV; American Psychiatric Association, 1994) include a number of features designed to enhance its cross-cultural applicability. However, the overt move toward a culture-sensitive nosology is undermined by an implicit assumption of the universality of its primary syndromes. In this review we argue that the DSM-IV's underlying thesis of universality based on Western-delineated mental disorders is problematic and has limited cross-cultural applicability. Research on the cross-cultural manifestation of schizophrenia and depression shows that presentation of these disorders varies significantly across cultures. We conclude by discussing the research and clinical implications of these findings. PMID- 9740977 TI - The functions of self-mutilation. AB - While pathological self-mutilating behavior has been clinically examined for over 65 years, and much of the literature hypothesizes some function for the behavior, there has been little attempt to integrate or differentiate between different functional ideas. This review uses six functional models extracted from the literature to organize a discussion of the multiple functions of self-mutilation, acknowledging the overdetermined nature of the behavior and attempting to understand how self-mutilation can serve multiple functions simultaneously. Contextual information about the definition, prevalence, phenomenology, patient characteristics, associated diagnoses, and associated symptoms of self-mutilation is first presented. Six functional models are then presented: the environmental model, the antisuicide model, the sexual model, the affect regulation model, the dissociation model, and the boundaries model. Support for these models in the empirical and theoretical literature is presented and treatment implications are explored. PMID- 9740978 TI - A review of methods and instruments for assessing externalizing disorders: theoretical and practical considerations in rendering a diagnosis. AB - This review addresses the most current and widely used methods of assessing childhood and adolescent externalizing disorders. Interviews, rating scales, and self-report instruments are described, and their strengths and weaknesses are discussed. Direct observational techniques in naturalistic and analogue settings are also reviewed. Throughout the article, commentary is offered regarding the psychometric adequacy and clinical validity of these instruments. It is suggested that, although the instruments presently used to assist in diagnosing externalizing disorders generally possess adequate reliability and representational validity, evidence of elaborative validity is lacking. Clinicians and researchers are encouraged to adopt a broader conceptualization of the diagnostic process, to question existing standards for establishing validity, and to consider alternative means of demonstrating diagnostic utility. PMID- 9740979 TI - Evolution of personality disorder diagnosis in the Diagnostic and Statistical Manual of Mental Disorders. AB - This article reviews the history and evolution of the diagnosis of personality disorders according to the Diagnostic and Statistical Manual of Mental Disorders (DSM) from its first edition in 1952 through its fourth edition in 1994. The article also traces the earliest origins of personality disorders (e.g., Hippocrates) through the modern foundational works of Pritchard, Schneider, and Horney. Analysis of the changes across the editions of the DSM suggest slow but steady progress in the clarification and classification of personality disorders, although formidable challenges remain. A call for future research as to reliability and validity of personality disorders is made, and suggestions for research are offered. PMID- 9740980 TI - Chronic fatigue syndrome: Its cause and a strategy for management. AB - This article describes the features of chronic fatigue syndrome and, by analysis of the many clinical paradoxes which it manifests, attempts to give a unifying explanation of the cause of the disorder and a strategy for management. PMID- 9740981 TI - Answers to 10 key questions on diverticular disease of the colon. AB - Diverticular disease is common in industrialized countries, and will become more prevalent in the future. Although it is usually a benign condition, treatment with high-fiber diet may prevent complications such as infection, stricture, or bleeding. PMID- 9740982 TI - Clinical evaluation of low back pain. AB - To properly diagnose and treat low back pain, a thorough history and physical examination are the cornerstones. The most important diagnoses for the physician to be aware of are cauda equina syndrome, back strain, herniated disc, stenosis, and spondylolisthesis. PMID- 9740983 TI - Inhaled steroids in asthma. AB - Inhaled steroids are the mainstay of anti-inflammatory treatment of asthma. Topical application of steroids to the asthmatic airway has many benefits as well as potential side effects; however, the side effects are much less prevalent than those caused by the use of chronic oral prednisone. PMID- 9740984 TI - Operative arthroscopy into the next century. AB - Until 10 years ago, operative arthroscopy was largely restricted to the knee. However, shoulder arthroscopy can now stabilize instability and repair rotator cuff tears. Operative procedures are seven being extended into the wrist and ankle. PMID- 9740985 TI - National Diabetes Education Program releases guiding principles for diabetes care for health care providers. PMID- 9740986 TI - A rational approach to the outpatient management of lacerations in pediatric patients. AB - Lacerations are a frequent reason for pediatric health care visits. Many are referred to EDs or to surgical specialists but may be treated by the pediatrician who has the time and interest in maintaining wound care skills. Although skin closure is often viewed as the primary event in wound care, local anesthesia and wound toilet are equally important aspects in which expertise is often undervalued. On occasion, patient anxiety and resistance complicates wound care, and a variety of sedative techniques facilitates completion of procedures that otherwise would require general anesthesia. Adherence to basic principles and the occasional use of innovations in wound care enable the clinician to bring about optimal outcomes. PMID- 9740987 TI - Endometrial cytology in postmenopausal hormone replacement therapy. AB - Endometrial changes is postmenopausal hormone replacement therapy (HRT) were studied by comparing cytological and histological findings. Cytological and histological examinations were conducted on 138 benign cases and 26 abnormal cases, including 24 cases with disordered proliferative phase (DOP) and 2 cases with simple endometrial hyperplasia (SEH), for a total of 164 cases. Hormones were administered as follows: 1) single cyclic administration of estrogen only (Single-HRT) for 31 cases, 2) cyclic administration of estrogen and progestin (Cyclic-HRT) for 105 cases, and 3) continuous administration of estrogen and progestin (Continuous-HRT) for 28 cases. All of the 164 cases were studied cytologically as to shape, appearance, nuclear number on maximum diameter, and so on. The benign cases in each mode of administration as described above revealed the following: 1) Single-HRT, atrophy or the proliferative phase was noted histologically, and the copresence of the endometrial epithelium and the ciliated cell metaplasia was observed cytologically; 2) Cyclic-HRT, the first half of the administration term was of the proliferative phase histologically, and the linear and long glands were seen cytologically. In the latter half of the administration term the secretory phase was noted histologically and the curved/linear glands with subnuclear vacuolization were observed cytologically; and 3) Continuous-HRT, atrophy was noted histologically, and fewer glands and atrophic cells on the endometrial epithelium with wrinkles mixed therein were seen cytologically. On the other hand, cytological examinations of the abnormal cases revealed a mean average of 35 nuclei on the maximum diameter of the gland, protrusion and/or ramification of the glands, densely clustered glands, and back-to-back glands without fusion, as well as irregularly dilated tortuous glands in SEH. These abnormal findings were considered useful for early detection of endometrial disorders in the hormone replacement therapy by cytodiagnosis. PMID- 9740988 TI - Fine-needle aspiration cytology of metastatic nasopharyngeal carcinoma. AB - Over a 32-month period at the University Hospital, Kuala Lumpur, we were able to study the cytological appearance of metastatic nasopharyngeal carcinoma (NPC) in 17 cases. This comprised 14 males and three females of which 13 were Chinese, three were Malay, and one was Indian. Their ages ranged from 27 to 64 years. Histological correlation was available in all the patients in the form of nasopharyngeal biopsies, and they were classified as per the World Health Organization classification into types I, II, and III NPC. Smears from type II NPC showed good cellularity with mainly clustered and occasionally dissociated cells, with focal columnar appearance, vesicular nuclei, prominent nucleoli, and variable amounts of cytoplasm. Clusters of malignant cell closely associated with lymphoid cells and dissociation of malignant cells were more characteristic of type III NPC. FNA cytology is now applied extensively to the diagnosis of head and neck tumours and knowledge of the cytomorphology of NPC would greatly aid in pinpointing the primary of this tumour which is notorious for presenting with early nodal metastasis. PMID- 9740989 TI - Cytologic features of mycobacterial pleuritis: logistic regression and statistical analysis of a blinded, case-controlled study. AB - Tuberculous pleural effusions are characterized by lymphocytosis; the significance of mesothelial cells is uncertain, as are the cytologic features in concurrent human immunodeficiency virus (HIV) infection. This blinded study compared 38 culture-positive pleural fluids (6 HIV+) with 38 controls from benign exudative processes. Logistic regression analysis selected mature lymphocytes as most predictive of positive culture, and mesothelial cells and eosinophils as negative predictors. Mesothelial cells were scant (< 10% of nucleated cells) in 36/38 cases with mycobacteria (sensitivity 95%); if these cell were > 10%, tuberculosis was virtually ruled out in HIV- patients. Specificity was maximized (82%) when mesothelial cells < 10% were combined with lymphocytes > 50%; positive predictive value with this combination was 76%, but was raised to 96% if moderate/marked cellularity was also identified. Among tuberculosis cases, reactive mesothelial cells differentiated HIV+ from HIV- patients; there was no other significant difference. PMID- 9740990 TI - Fine-needle aspiration biopsy of metastatic small cell carcinoma from extrapulmonary sites. AB - Like a pulmonary counterpart, extrapulmonary small cell carcinoma (SCC) is an aggressive tumor with a high rate of metastasis. Forty-nine fine-needle aspiration biopsies (FNABs) (36 patients) of various primary sites other than the lung diagnosed as metastatic SCC (including Merkel cell carcinoma) were reviewed. FNABs were derived from lymph nodes (20), liver (7), bone (2), breast (1), pancreas (1), and skin/soft tissue (18). Primary tumor sites included the prostate (14), skin (11; Merkel cell carcinoma), cervix (5), urinary bladder (3), urethra (1), ovary (1), and parotid (1). Aspirates revealed predominantly dispersed single tumor cells with occasional clustering. Tumor cells were small with scant cytoplasm, fine powdery chromatin, and inconspicuous nucleoli. Nuclear molding, mitotic figures, and apoptotic bodies were frequently observed. In four cases, findings from the FNABs were used to render the initial diagnosis of SCC. FNAB is useful for determining whether metastases contain a SCC component, a finding that may alter clinical management. Cytologically, SCC from different primary sites cannot be differentiated, and its distinction requires clinical and radiographic correlation. PMID- 9740991 TI - Cytologic features of "dry-type" cutaneous leishmaniasis. AB - Exfoliative cytology smears from the lesions of 179 patients with cutaneous leishmaniasis due to Leishmania tropica were studied with specific reference to cellular reactions and their effect on the parasite. Aggregates of the parasite (so-called Leishmania Donovan bodies) were present within macrophages and in some fibroblasts. The nature of the inflammatory reaction to the disease was studied by performing differential counts of the inflammatory cells present in the smears. These were correlated with the number of Leishman Donovan bodies. There was an inverse relationship between the number of Leishman Donovan bodies and the percentage of small lymphocytes, neutrophils, and type I macrophages. It is postulated that aggregates of activated macrophages (designated types II and III) and the Leishmanian milieu (sticky matrix) protect the amastigote Leishmania parasites from being eradicated by the inflammatory and immune reaction. The cytoplasmic blebbing of the parasitophorous vacuoles and cell to cell connection of the activated histiocytes could be shown by the CD-68 immunostaining of the tissue biopsy. PMID- 9740992 TI - Fine-needle aspiration cytology of an oncocytic carcinoma of the submandibular gland. AB - An oncocytic carcinoma of the submandibular gland diagnosed by fine-needle aspiration is reported. Cytologically, the tumor cells occurred singly, in three dimensional clusters and in multilayered sheets. The cells had round-to-oval, centrally or eccentrically located nuclei with fine chromatin and prominent nucleoli. Many cells contained abundant granular cytoplasm and well-defined cell borders; however, several stripped nuclei with prominent nucleoli were also noted in the background. Follow-up histologic examination showed tumor cells arising from right submandibular gland and metastasizing to six of 14 cervical lymph nodes. Ultrastructural study demonstrated tumor cells packed with mitochondria in the cytoplasm. The patient was alive with no evidence of disease 6 months after the surgery. PMID- 9740993 TI - Cytological aspiration of intraocular retinoblastoma in an 11-year-old boy. AB - Only a few cases of retinoblastomas in older children have been reported and the clinical diagnosis may be difficult. In case, fine-needle aspiration from an atypical retinal mass of an 11-yr-old boy was performed. The vitreous fluid was stained with Diff-Quik for an immediate cytological examination and the diagnosis of retinoblastoma was suggested. The rest of the specimen was separated into two parts. One was stained with May-Grunwald-Giemsa and the other was centrifuged, embedded in paraffin, and finally stained with hematoxylin-eosin-safran. The undifferentiated blue cells were associated with abundant necrotic debris and portions of capillaries with perivascular tumor cells around. The cytoplasm of the tumor cells was strongly stained with neuron-specific enolase antibody. The diagnosis of retinoblastoma was confirmed. The specimen of enucleation confirmed the diagnosis. In conclusion, cytological aspiration can categorically diagnose suspected intraocular tumors of older children in whom clinical and noninvasive investigations have failed to establish the diagnosis. PMID- 9740994 TI - Fine-needle aspiration cytology of metastatic clear-cell renal carcinoma presenting as a solitary mass in the head of the pancreas. AB - In nearly 25% of patients with primary renal-cell carcinoma, metastasis is not uncommon and usually does not give rise to difficulties in diagnosis. However, its presentation as a mass in the head of the pancreas following an elapse of several years after the initial diagnosis of renal carcinoma is not only uncommon but may be confused clinicoradiologically with a primary pancreatic adenocarcinoma. The case presented here illustrates such an example with an emphasis on the usefulness of fine-needle aspiration cytology (FNAC) in the diagnosis. PMID- 9740995 TI - Neuritic Hansen's disease: an unusual presentation and a diagnostic challenge on fine-needle aspirate. AB - Fine-needle aspiration cytology is now routinely used in the diagnosis of cervical lumps. We report on a case of leprous neuritis which presented as a cervical swelling. A diagnosis of leprosy was suggested on the basis of globi within histiocytes. These histiocytes differed from those classically described in leprosy. Retrospective review demonstrated the presence of occasional nerve fragments, and some of these closely resembled granulomas. The possible close resemblance of this lesion to tuberculosis and a histiocytic proliferative disorder is highlighted. Recognition of nerve fragments along with histiocytes should suggest the diagnosis. PMID- 9740996 TI - Salivary duct carcinoma ex pleomorphic adenoma: analysis of the findings in fine needle aspiration cytology and histology. AB - Salivary duct carcinoma (SDC) is an uncommon and highly aggressive neoplasm that reveals histopathologic features resembling infiltrating duct carcinoma of the breast and prostate. SDC arising on the basis of preexisting pleomorphic adenoma (PA) is very rare. This report describes the fine-needle aspiration (FNA) cytology of a case of SDC ex PA. FNA smears were cellular with cells with large, pleomorphic nuclei, some prominent nucleoli, and finely vacuolated cytoplasm. The neoplastic cells were forming groups, sheets, and papillary structures and a cribriform pattern was present in some sheets. Mitotic figures were frequently seen. Necrosis was prominent in the background. Histologic sections of the tumor revealed areas of comedo carcinoma, papillary carcinoma, and infiltrative carcinoma as well as multiple foci of PA. The presence of a cribriform pattern, tumor cells resembling breast ductal carcinoma cells, and a necrotic background are helpful features for an accurate cytodiagnosis of SDC. PMID- 9740997 TI - Fine-needle aspiration of large cell lymphoma in a patient with agnogenic myeloid metaplasia: a case report and review of the literature. AB - We describe a patient with agnogenic myeloid metaplasia and Sjogren's disease who developed a large B-cell lymphoma. We discuss the differential diagnoses of fine needle aspirations of a localized mass from such patients and review the literature of the association between agnogenic myeloid metaplasia and lymphoma. PMID- 9740998 TI - Fine-needle aspiration of basaloid squamous carcinoma: a case report with review of differential diagnostic considerations. AB - Basaloid squamous carcinoma is a distinct variant of squamous carcinoma with a particularly poor prognosis. To our knowledge, there are only two papers in the cytopathology literature which describe this entity. We report the fine-needle aspiration findings of an additional case of metastatic basaloid squamous carcinoma in a cervical lymph node and compare its cytomorphologic features to those observed on touch imprints of the subsequent surgical specimen. Smears of the aspirate showed a mixed lymphoid background with interspersed cohesive clusters of small cells roughly 3 times the size of small mature lymphocytes. Some cells were angulated and others exhibited irregular nuclear contours. The cells were generally hyperchromatic with evenly staining dense chromatin or irregularly distributed coarse chromatin. Focally there was evidence of nuclear molding. On Diff-Quik staining, irregular globules of magenta-stained extracellular dense material were noted within or adherent to the periphery of some clusters or as somewhat linear formations with small epithelial cells clinging to the edges. Abundant mitotic figures and clumps of necrotic tumor were more apparent on touch preps of the subsequent surgical specimen. The differential diagnosis by fine-needle aspiration includes adenoid cystic carcinoma, basal-cell adenocarcinoma, adenosquamous carcinoma, and small-cell carcinoma. If a fine-needle aspirate of a cervical lymph node shows the features described above and the primary tumor is unknown, suggesting the possibility of metastatic basaloid squamous carcinoma may aid clinicians in the search for a primary site, as basaloid squamous carcinoma occurs most frequently at the base of the tongue, hypopharynx, and supraglottic larynx. PMID- 9740999 TI - Transthoracic fine-needle aspiration cytology of inflammatory pseudotumor, fibrohistiocytic type: a case report with immunohistochemical studies. AB - Inflammatory pseudotumor, fibrohistiocytic type, also called benign fibrous histiocytoma, is a rare reactive entity usually found incidentally on routine chest roentgenography. We present a case of inflammatory pseudotumor, fibrohistiocytic type, initially diagnosed by fine-needle aspiration (FNA) cytology in a 39-yr-old woman with a history of breast carcinoma. Cytomorphologic characteristics were confirmed by a cell block examination and immunohistochemical findings. The differential diagnoses of the fine-needle aspiration cytology of this type of inflammatory pseudotumor are discussed. PMID- 9741000 TI - PSA immunoreactivity in a parotid oncocytoma: a diagnostic pitfall in discriminating primary parotid neoplasms from metastatic prostate cancer. AB - Prostate-specific antigen (PSA) is secreted by both normal and neoplastic acinar cells of the prostate gland, and the immunohistochemical detection of PSA is widely accepted as an excellent method for confirming the prostatic origin of metastatic tumor implants in men with prostate cancer. Less recognized is the observation that certain nonprostatic tissues and their neoplastic counterparts also secrete PSA. As one example, salivary gland ducts and certain salivary gland neoplasms have been reported to be immunoreactive for PSA. Potentially, this nonspecificity could be a diagnostic pitfall when using immunoperoxidase on fine needle aspiration (FNA) biopsy specimens to differentiate metastatic prostate cancer from primary salivary gland tumors. We report on a case where strong PSA immunoreactivity of a parotid oncocytoma led to its confusion with metastatic prostate cancer. PMID- 9741001 TI - Positive stain for mucin in nipple discharge fluid--a mucin-producing lesion or not? PMID- 9741002 TI - Some functional observations related to the N18 component of the median nerve SEP. AB - This work describes the results of two experimental procedures related to the short latency median nerve somatosensory evoked potentials. In the first one, it was possible to show that there is a different rate of increase in the amplitude of the different components related to different intensities of stimulation. In the second one, the simultaneous stimulation of the median and the ulnar nerves disclosed an occlusive interaction which was larger for the N18 than for the other components. These results may be a support to the hypothesis that the N18 component generator is related to the mechanism of presynaptic inhibition within the cuneiform nucleus. PMID- 9741003 TI - Initial motor unit recruitment in patients with spastic hemiplegia. AB - Lesions of the first motor neuron provoke abnormal voluntary movements. To clarify the central nervous system mechanisms underlying these changes we analyzed the behavior of the first motor unit recruited during a minimal effort tonic contraction of the deltoid and abductor digiti minimi manus muscles in patients with hemiplegia due to cerebrovascular lesions in the distribution of the middle cerebral artery. We compared data from paretic and healthy muscles in the same subject. The onset and recruitment intervals determined for the single motor unit yielded the range of control. The first recruited motor unit had a lower baseline firing rate and the second recruited motor unit potential appeared significantly earlier (p < 0.01) in plegic than in healthy muscles. Both changes affected distal more than proximal locations. Recordings from plegic muscles, particularly at distal locations, also disclosed a lowered range of control. These findings suggest that in hemiplegic patients the central nervous system loses its ability to modulate the frequency of firing during minimal effort voluntary movements so that distal muscles tend to behave like proximal muscles. PMID- 9741004 TI - Tonic stretch reflexes in older able-bodied people. AB - It is a general assumption that, in able-bodied persons, tonic stretch reflex (TSR) activity is not elicited during stretching of relaxed muscles and that the presence of TSR activity following brain damage is, therefore, indicative of spasticity. However, a variety of studies have reported age-related changes in reflex activity, raising the question of whether this assumption is justified in older subjects. The aim of this study was to determine if TSRs were activated in the relaxed elbow flexors of able-bodied people in an age-group at risk of stroke. Electromyographic (EMG) activity was recorded in 30 able-bodied subjects aged 46 to 78 years when their relaxed elbow flexors were subjected to ramp and sinusoidal stretches of different amplitudes and velocities. It was found that these subjects did not exhibit TSR activity under these conditions. Therefore, the practice of measuring TSR activity as a means of quantifying spasticity in stroke patients appears justified. PMID- 9741005 TI - Optimal electrode placement in facial nerve conduction study. AB - This study was designed to determine the optimal sites for the active electrode in a nerve conduction study of each branch of the facial nerve. Twenty healthy male and female volunteers between 20 and 40 years old were investigated. Our criteria for the optimal site of the active electrode were initial negative deflection and maximal amplitude of the response and the most synchronized response. Optimal sites were found to be as follows: 1. Frontalis (temporal branch): a point midway between the hairline and the eyebrow along a line passing vertically through the pupil. 2. Orbicularis oculi (zygomatic branch): the medial quarter between the medial and lateral canthus. 3. Nasalis (buccal branch): muscle belly. 4. Triangularis (mandibular branch): 15 mm lateral and 25 mm below the corner of the mouth. 5. Orbicularis oris (zygomatic, mandibular and buccal branches): 2 mm below the lower lip midway between the midline and the corner of the mouth. PMID- 9741006 TI - Effect of voluntary muscle contraction on the startle response to auditory stimuli. AB - Startle reflex responses were studied in 15 normal human subjects using weak (88 dB) and strong (114 dB) auditory stimuli in the orbicularis oculi, masseter, sternocleidomastoid, trapezius, deltoid, biceps, forearm flexors and quadriceps muscles. With the subjects in the relaxed state, no consistent responses were seen with the weak stimuli, and with the strong stimuli responses were only present in orbicularis oculi muscles. When the above muscles were in a state of voluntary contraction, the strong stimuli produced complex responses which were not always excitatory in nature, with muscle relaxation being noted in a number of stimulation sequences. Repetitive weak and strong stimuli were used to study habituation effects in the orbicularis oculi muscles. The repetitive strong stimuli produced a wide range of response patterns, indicating a high inter individual variability in habituation. In two subjects, no habituation effects were present. Our study supports the high intra-individual variability of the startle response, and suggests that this response is affected by the state of muscle contraction at the time of stimulation. Startle response is more easily elicited in a state of muscular contraction. Future studies of startle reflex should take this into consideration. PMID- 9741007 TI - Reproducibility of different methods for diagnosing and monitoring diabetic neuropathy. AB - Conflicting results have been published concerning the reliability of methods to assess diabetic polyneuropathy. Therefore we compared the reproducibility of nerve conduction studies and quantitative measurements of vibratory sensation in 23 diabetic patients with clinically and/or neurophysiologically established neuropathy. Motor nerve conduction of peroneal and median nerves, sensory nerve conduction of sural and median nerves and vibratory perception thresholds of index fingers and big toes (forced-choice method) were assessed on three occasions with an interval of one to fourteen days, within a period of three weeks. We conclude that both nerve conduction studies and vibratory perception threshold measurements are reproducible methods to assess diabetic neuropathy. Determination of vibration perception thresholds has the advantage of being a simple and unobtrusive method. Nerve conduction studies are less variable and therefore more suited for monitoring diabetic neuropathy. PMID- 9741008 TI - An experimental study about the incorrect electrophysiological evaluation following peripheral nerve injury and repair. AB - OBJECTIVE: This experiment was designed to explore the reasons of incorrect electrophysiological evaluation following peripheral nerve repair. METHODS: Twenty-four New Zealand white rabbits were used and divided into 2 groups. The first group of animals underwent severance of gastrocnemius muscle nerve and only the medial branch of the nerve was anastomosed. The second group of rabbits underwent severance of the muscle nerve as well as sural nerve, then the proximal end of the sural nerve was anastomosed to the distal end of the muscle nerve. Electrophysiological and morphological methods were used to study these models 3 months after the operations. RESULTS: Electrophysiologic comparison of muscle action potential for group A and B showed a significant difference in the values (p < 0.05). Morphologic studies showed that the muscle weights of group B were significantly lower than those of group A, but the myelin thickness failed to show any statistical difference between the two groups. CONCLUSION: Results suggested that one of the sources causing incorrect electrophysiological evaluation could be misdirected regeneration, resulting from a sensory fascicle anastomosing to a motor one or vice versa. The unfunctional regeneration showed low values of muscle action potentials (p < 0.05) as well as excessive fibrillation potentials, and marked atrophy of the muscles (p < 0.05). PMID- 9741009 TI - Normal values of F wave in upper extremities of 50 healthy individuals in Iran. AB - F wave latency has been shown to be a valuable method in evaluation of a variety of neurologic disorders. We measured F wave values in 50 healthy individuals in Shiraz. Maximum normal F wave latency for median nerve was 25.7 ms for women and 28.5 ms for men with stimulation at the wrist. It was 23 ms for women and 25 ms for men with stimulation at the elbow. Maximum normal F wave latency for ulnar nerve was 26.45 ms for women and 28.9 ms for men with stimulation at the wrist. It was 23.1 ms for women and 25.3 ms for men with stimulation at the elbow. Maximum normal difference in F wave latency between right and left upper extremities with stimulation at the wrist for total group was 2.2 ms for median nerve and 2.4 ms for ulnar nerve. Maximum normal difference in F wave latency between median and ulnar nerve in an extremity with stimulation at the wrist for total group was 2.7 ms. There was statistically significant difference in F wave latency between women and men. PMID- 9741010 TI - Magnetic stimulation of the peripheral nervous system: local versus generalized disorders. AB - Magnetic stimulation was used to excite lumbar and cervical roots, enabling the measurement of proximal motor conduction besides the routine measurement of distal conduction velocity (CV). Latencies to abductor pollicis brevis (APB) and tibial anterior (TA) on both sides were used to estimate peripheral conduction velocity using body height; in addition, the uniformity of conduction was determined using the ratio of lumbar and cervical latencies (L/C ratio). In 124 controls the CV showed a clear inverse relation with age. After correction for this factor a high correlation between left and right CV remained. The L/C ratio- indicating the uniformity of conduction--was plotted against the CV, which made it possible to view both generalized and focal slowing in one diagram. An application of this approach is shown in patients with generalized slowing (demyelinating polyneuropathy), focal slowing (lumbar stenosis) and multifocal slowing (acute and chronic inflammatory demyelinating polyneuropathy). Most disorders could be accurately deduced from their place in the diagram. A special variable (distance between left and right data) was introduced to characterize the special case of multifocal slowing. It is concluded that this method easily provides an idea of global and/or focal peripheral conduction slowing and that it might be useful besides routine EMG as an additional tool. PMID- 9741011 TI - Compound muscle action potential amplitude and area changes in normal subjects and patients with carpal tunnel syndrome. AB - Lesions of mixed peripheral nerves are often diagnosed by means of electrodiagnostic tests, especially by motor and sensory conduction studies. In order to clarify the varying degrees of motor nerve involvement in patients with carpal tunnel syndrome (CTS), we designed this study for comparing the results of compound muscle action potential (CMAP) amplitude and area under the negative phase between normal persons (n = 662) and patients having CTS (n = 250). The CTS cases were categorized into two distinct groups, i.e., CTS-D (n = 120) having prolonged median sensory latency and normal needle examination, CTS-AD having prolonged median sensory latency and abnormal needle findings (n = 130). Subjects in the CTS-AD group showed significant reduction, both in CMAP amplitude and area from 3rd to 7th decades; however, in the CTS-D group there were significantly smaller CMAP amplitude from 5th to 7th decades but CMAP area demonstrated wide variations. These data indicate that CMAP amplitude and area are helpful in documenting motor nerve axonal loss, but care must be exercised when attempt is made to define axonal loss based either on area or amplitude because of the temporal dispersive effects. PMID- 9741013 TI - [Hypnogogic hallucinations of macrosomatognosia and microsomatognosia]. AB - Macrosomatognosia and microsomatognosia, in which the entire body or parts of the body are perceived as abnormally large or small, can occur as hypnagogic hallucinations in sane and healthy subjects. A review of the literature and five original observations are presented. The mouth and the hands are anatomic regions which are frequently affected, paralleling the dominance of their representation in the sensory maps of the human brain. The clinical differential diagnosis of the said phenomena includes narcolepsy, epileptic and migraine aura, drug-induced body schema disturbances and basic symptoms of functional psychoses. PMID- 9741014 TI - [Classical German brain pathology especially aphasiology--The Berlin Charity hospital]. AB - The history of the classical period of German clinical brain pathology, especially aphasiology, is described. The present article covers authors, neuropsychiatrists and one phoniatrist, all of whom worked in Berlin before the First World War. The study aims at providing an insight into that most fruitful period to those who are presently concerned with assessment and treatment of aphasia and other neuropsychological disorders. PMID- 9741015 TI - [Elecroencephalographic charactistics of Creutzfeldt-Jakob disease and its differential diagnosis]. AB - Although clinical electroencephalography is no longer as important as it used to be in differential diagnosis of a fair number of neurological and psychiatric diseases ever since imaging techniques have been making enormous strides, EEG is still an important diagnostic tool in dementias where specific morphological lesions are not immediately or not at all apparent which would otherwise be visible by imaging. Sporadic Creutzfeldt-Jakob disease is an important case in point. Although this is associated with some unspecific EEG findings, typical periodical sharp wave complexes (PSWC) become conspicuous in the course of the disease. If these are meticulously studied and particular attention is paid to their periodicity, a sensitivity of 67% and a specificity of 86% are attained. With the exception of one familial variant of Creutzfeldt-Jakob disease PSWC ar usually absent all other human prion diseases. Hence, it is not likely that they are linked to the aetiology of sporadic Creutzfeldt-Jakob disease. We present a patho-physiological hypothesis on the development of PSWC basing on the assumption that the specific periodicity of PSWC results from a still functionally active but greatly impaired subcortical-cortical circuit of neuronal excitability. This specific pattern of neuronal degeneration may obviously arise- albeit very rarely--also in other diseases independent of their aetiology, so that the EEG patterns appear identical. For this reason it is imperative to make complementary use of EEG and of recent clinical and laboratory data of Creutzfeldt-Jakob disease before PSWC and be considered a relevant diagnostic criterion. Conversely, clinical diagnosis of Creutzfeldt-Jakob disease should be reconsidered if repeated EEG recordings fail to reveal PSWC even under technically adequate conditions. PMID- 9741016 TI - [From fighting to preventing disease. Is such a paradigm possible for schizophrenic disorders?]. AB - Research on early detection and early treatment of schizophrenia is not only based on findings of studies of course, outcome and therapy, but is the most obvious consequence of the nowadays dominating conceptualisation of schizophrenia. In the last decades, results of birth cohort studies as well as of genetic and psychometric "high-risk" research, of epidemiological studies and of studies of the clinical course have given starting points for the development of early detection and early intervention programmes. Such programmes will be applicable most likely at the begin of the psychotic first manifestation, but it seems possible that they can be extended to the prodromal phase or the premorbide phase. Therefore, it is important to build up early detection networks based on regional early detection centres interacting between each other via an international network. The main aim of these centres--once sensitive screening instruments are existent--should be the identification of not yet psychotic at risk persons in the general population and, after the application of specific prediction instruments, their preventive treatment. PMID- 9741017 TI - [Self-experienced vulnerability, prodromic symptoms and coping strategies before schizophrenic and affective episodes]. AB - For the first time, the present study explores self-experienced vulnerability, prodromal symptoms and coping strategies preceding schizophrenic and affective episodes. 33 schizophrenic and 29 depressive patients were assessed retrospectively for preepisodic alterations by means of the "Bonn Scale for the Assessment of Basic Symptoms- BSABS" after complete recovery from the acute episode. 97% of the schizophrenic and 93% of the depressive patients showed preepisodic alterations. In the schizophrenic group the first alteration occurred with a median of 10 weeks and in the depressive group with a median of 18 weeks before the onset of the acute episode. With regard to self-experienced vulnerability depressive cases were significantly less tolerant to stress, i.e work under time pressure or unusual, unexpected requirements. With regard to prodromal symptoms schizophrenics showed significantly more often interpersonal irritation and certain perception and thought disturbances, whereas depressive patients reported more often adynamia and certain disturbances of proprioception. 73% of the schizophrenic patients and 90% of the depressive patients reacted to early symptoms with coping strategies. The preepisodic alterations in schizophrenic patients could be described in terms of mild psychotic productivity, early symptoms of depressive patients could be described as a mild depressive syndrome. Prospective studies are necessary to show if assessment of mild psychotic productivity could be used for early diagnosis and early intervention in schizophrenia. PMID- 9741018 TI - Intravenous corticotrophin vs. hydrocortisone in the treatment of hospitalized patients with Crohn's disease: a randomized double-blind study and follow-up. AB - Adrenocorticotrophic hormone (ACTH) and corticosteroids have no maintenance values for inflammatory bowel disease but serve to reduce the severity of disease. The effectiveness of intravenous corticotrophin versus hydrocortisone in ulcerative colitis has been determined including whether previous steroid therapy influenced the better response to one rather than the other, but no such studies have ever been done in Crohn's disease. Eighty-eight patients hospitalized with moderate-to-severe Crohn's disease (Present-Korelitz [P-K] Index -3 to -2 and the International Organisation for the Study of Inflammatory Bowel Disease-Crohn's & Colitis Foundation of America [IOIBD-CCFA] Index, mean 14, range 5-23) were treated in a prospective, randomized, double-blind clinical trial to receive either continuous intravenous infusion of 120 U/day of ACTH (44 patients) or hydrocortisone 300 mg/day (44 patients). Patients were also subdivided into those who received oral steroids during the 30 days prior to intravenous therapy and those who had not. Response was followed on a daily basis and tabulated at 3, 5, and 10 days. Patients were followed from 1-3 years to determine the later status. After 10 days of intravenous therapy 36 of 44 patients (82%) who received ACTH and 41 of 44 patients (93%) who received hydrocortisone fully responded (P-K index +3 and IOIBD-CCFA Index mean of 3). At the end of the study, response to intravenous ACTH and hydrocortisone was not statistically different whether or not patients received oral steroids during the 30 days prior to admission, although the response to IV ACTH tended to be faster at 3 days in those who had received previous steroid therapy. Intravenous ACTH and hydrocortisone are equally effective in achieving therapeutic goals in patients with Crohn's disease who have not achieved results with oral medications. Moreover the response rate was high (mean 88%), serving to buy time for establishment of successful maintenance programs of treatment with oral 5-ASA and immunosuppressive drugs for 69% of patients at 1-3 years. PMID- 9741019 TI - Inflammatory bowel disease: is there any relation between smoking status and disease presentation? European Collaborative IBD Study Group. AB - Smoking is associated with Crohn's disease and nonsmoking with ulcerative colitis. The aim of this study was to compare the clinical features at diagnosis and during the first year of follow-up in smokers and nonsmokers with inflammatory bowel disease (IBD). In 19 centers across Europe, a prospective study was performed of 457 newly diagnosed patients with Crohn's disease and 930 with ulcerative colitis. The characteristics of the disease were recorded by the treating physician by using a standard protocol at the time of diagnosis. Treatment characteristics were assessed after 1 year of follow-up. Weight loss occurred significantly more often in smoking patients with Crohn's disease, as well as in smokers with ulcerative colitis (p < 0.02), and diarrhea was more frequent in smoking patients with Crohn's disease compared with non-smoking individuals (p < 0.01). Patients with Crohn's disease who smoke were less likely to have colonic involvement (p < 0.01) and were more often prescribed immunosuppressive medication (p < 0.02). The study suggests that (a) smoking protects the colon from inflammation and (b) is associated with more active disease in Crohn's disease. The association between weight loss and smoking in both diseases is probably due to a general effect of smoking. The reported relation between smoking and the course of Crohn's disease is a strong argument for encouraging patients to give up smoking. PMID- 9741020 TI - A rat model of ileal pouch-rectal anastomosis. AB - After colectomy and ileal pouch-rectal anastomosis, pouchitis may occur. Pouchitis is a poorly defined condition with unknown etiology. The aim of this study was to develop an animal model of pouchitis. Ileal pouch-rectal anastomosis was created in Lewis and Sprague-Dawley rats. Rats were studied 4 and 8 weeks after surgery, and pouchitis was assessed by stool output, histology, and tissue myeloperoxidase (MPO) levels. Some rats were treated with allopurinol or metronidazole beginning the day of surgery. Rats with pouches demonstrated inflammation with a monocytic infiltration, luminal exudate, mucosal ulcerations, and serosal inflammation. Rats with pouches had increased anaerobic bacterial flora compared with normal ileum. After creation of pouches, Lewis rats (histology score = 8.4 +/- 1.6; MPO = 17.3 +/- 3.6, mean +/- SD) developed more severe inflammation than Sprague-Dawley rats did (histology score = 4.3 +/- 1.8; MPO = 5.5 +/- 3.6) within 4 weeks, p < 0.001 and 8 weeks after surgery, p < 0.05. Stool output was also greater in Lewis (55 +/- 7 g/kg/day) compared with Sprague Dawley rats with pouches (43 +/- 5 g/kg/day), p < 0.05. Metronidazole treatment reduced histology score (6.0 +/- 0.5) p < 0.05 and MPO (5.9 +/- 1.6) p < 0.001 in rats with pouches compared with rats with pouches that had no treatment. Allopurinol treatment in rats with pouches reduced histology score (4.0 +/- 1.7) and MPO (3.9 +/- 1.6), p < 0.001, compared with rats with pouches that had no treatment. Ileal pouch-rectal anastomosis in rats induced inflammation within 4 weeks, demonstrated differential host genetic susceptibility, and was associated with increased number of pouch bacteria. Anaerobes, especially bacteroides sp. and free radical, may mediate inflammation. Ileal pouch-rectal anastomosis surgery in rats may be a useful animal model for the study pouchitis. PMID- 9741021 TI - Development of colonic adenocarcinomas in a mouse model of ulcerative colitis. AB - Mice deficient in both interleukin-2 and beta 2-microglobulin expression (Beta 2mullnull x IL-2null mice) develop an inflammatory disease of the colon resembling ulcerative colitis. To examine long-term complications of disease in these mice, a group of 34 Beta 2mnull x IL-2null mice was monitored for 6-12 months. Development of clinical disease was assessed by wasting, general appearance, and diarrhea. Further analysis included histologic examination of the distal colon for colitis, staining of CD4+ T cells for surface activation markers, and cytoplasmic staining of CD4+ T cells for IFN-gamma and TNF-alpha. These older Beta 2mnull x IL-2null mice had activated CD4+ T cells as assessed by surface markers on flow cytometry. Cytoplasmic staining revealed IFN-gamma production, but not TNF-alpha production by CD4+ T cells. The majority of these older Beta 2mnull x IL-2null mice continued to have colitis on histology. However, they lived much longer and had less wasting in comparison to IL-2null mice. At necropsy, 11 (32%) of 34 of the Beta 2mnull x IL-2null mice had tumors in the proximal half of the colon. Histologic examination confirmed these tumors to be adenocarcinomas. These mice may be useful as a model for studying carcinogenesis in chronic colitis. PMID- 9741022 TI - Etiopathogenesis of inflammatory bowel disease: a pediatric perspective. PMID- 9741023 TI - Initial maintenance therapy (relapse prevention) and therapy of steroid resistance and dependence. PMID- 9741024 TI - Perianal disease, growth failure, and quality of life. PMID- 9741025 TI - Malignancy and aneuploidy: prevention and early detection. PMID- 9741026 TI - Hepatobiliary complications of inflammatory bowel disease: overview of the issues. PMID- 9741027 TI - Management of severe colitis/ileocolitis. PMID- 9741028 TI - Reliability of a Crohn's disease clinical classification scheme based on disease behavior. AB - Classification of Crohn's disease (CD) by disease behavior--either inflammatory (INF), fibrostenotic (FS), or fistulizing/perforating (FP)--has been proposed as a means of assisting management decisions and predicting outcomes for subgroup analysis in clinical trials and for making phenotype/genotype associations in molecular genetic studies. Accurate and reproducible classification of CD patient subgroups is of paramount importance in such studies but to be useful, the classification scheme must have good interrater agreement. We sought to assess the interrater agreement associated with the disease-behavior classification scheme of CD. Twelve patients with CD were randomly selected from a database of 964 patients with CD undergoing medical or surgical treatment or both. Clinical details of the 12 cases, along with their radiographs and surgical and pathological reports, were presented to a panel of 20 experts who were asked to classify each case based on the patient's overall disease course (scenario A) and as if the patient were being entered into a clinical trial on that day (scenario B). Calculations of strength of interrater agreement were made and were expressed as the kappa statistic (kappa), with kappa < 0.2 = poor strength of agreement; kappa 0.21 - 0.4 = fair; kappa 0.41 - 0.6 = moderate; kappa 0.61 - 0.8 = good; and kappa 0.81 - 1.0 = very good. Five panel participants did not complete the study, and three clinical vignettes were excluded because of incomplete scoring, leaving a total of 15 panel experts assessing nine cases. Overall interrater agreement was only fair with kappa = 0.353 for scenario A and kappa = 0.291 for scenario B. Interrater agreement was less when only the most straightforward case in each disease category was evaluated. Classification of CD by pattern of disease behavior yields only fair interrater agreement. This raises concerns regarding its applicability, particularly in ongoing studies of genotype/phenotype associations. Further refinement of disease subtypes and clear operational definitions are required. PMID- 9741029 TI - Risk factors and distinguishing features of cancer in IBD. PMID- 9741030 TI - Which immunosuppressors do you use to treat Crohn's disease and ulcerative colitis? In which order of priority and how worried are you about toxicity? PMID- 9741031 TI - Which immunosuppressors do you use to treat Crohn's disease and ulcerative colitis? In which order of priority and how worried are you about toxicity? AB - In severe and steroid-refractory UC, intravenous cyclosporin A is an effective tool to induce rapid remission. Short-term side effects can be dealt with, and decisions on further treatment can be made without emergency. AZA/6-MP can be used in steroid-dependent or chronically active disease and probably to maintain cyclosporin A-induced remission if surgery is not possible or not accepted. The latter should be discussed with the patient and an experienced surgeon. Other immunosuppressors do not play a role in my approach to this disease. In CD, AZA/6 MP is useful for steroid-refractory and -dependent patients. Again toxicity is limited and can be handled. This treatment should be used more frequently and is my approach of choice if fibrous stenoses, as cause of chronic activity, are excluded. MTX can be used as second-line treatment. I am worried about its long term toxicity, in particular to the liver, as described in rheumatic disorders. Both intravenous cyclosporin A and AZA/6-MP should be used by those having experience with these drugs (and IBD). They are much needed and helpful. PMID- 9741032 TI - Coding for screening and surveillance of colorectal cancer in the Medicare population. PMID- 9741033 TI - Pouch dysplasia: a new challenge. PMID- 9741034 TI - Structured Interview for Anorexic and Bulimic disorders for DSM-IV and ICD-10: updated (third) revision. AB - OBJECTIVES: Earlier versions of the Structured Interview for Anorexic and Bulimic Disorders (SIAB) were modified in order to include new research findings and to update the expert rating interview to the diagnostic criteria of DSM-IV and ICD 10. The semistandardized interview was developed for reliable and valid assessment of the specific as well as the general psychopathology of eating disorders. METHOD: Data from SIAB-EX interviews (current and past/lifetime symptom expression) were available from three samples: (a) 330 eating-disordered patients assessed at the start of treatment, (b) 148 former eating-disordered patients with anorexia nervosa (AN) or bulimia nervosa (BN) assessed at follow up, and (c) 111 community controls. Sixty-one of the 87 items of the SIAB-EX with a 5-point scale were factor analyzed. RESULTS: Principal components analyses with varimax rotation produced the following six components of the SIAB-EX (lifetime): (I) Body Image and Slimness Ideal; (II) General Psychopathology; (III) Sexuality and Social Integration; (IV) Bulimic Symptoms; (V) Measures to Counteract Weight Gain, Fasting, and Substance Abuse; and (VI) Atypical Binges. The factor solution for the current symptom expression was very similar to that based on lifetime symptom expression. Average item and factor scores are given for six groups of eating-disordered patients and controls. High interrater reliability was established for both current and the past symptom expression. Cronbach's alpha coefficients indicated good internal consistency for five of the six components of the SIAB-EX. DSM-IV and ICD-10 diagnoses for eating disorders can be derived directly or by using a computer algorithm from the SIAB-EX. A detailed 90-page manual facilitates the training of interviewers. CONCLUSION: The 87-item SIAB-EX was originally developed for detailed assessment of eating disorders cross sectionally and longitudinally. The updated version which allows for diagnosis according to DSM-IV and ICD-10 is described here. PMID- 9741035 TI - Binge eating disorder with and without a history of purging symptoms. AB - OBJECTIVE: The purpose of this study was to evaluate whether a history of purging behaviors in individuals with binge eating disorder (BED) is associated with increased comorbid psychopathology, dietary restraint, severity of eating pathology, and attitudinal disturbance in self-esteem and body image. METHOD: Sixty-three women meeting DSM-IV criteria for BED who were participating in a psychotherapy treatment study were subclassified according to whether they reported a history of purging behavior using self-induced vomiting or laxatives (HP; N = 24) or no such history (NHP; N = 39). The two groups were compared on the following variables: DSM-IV Axis I Lifetime diagnoses, Hamilton Depression Rating Scale, Body Shape Questionnaire, Three Factor Eating Questionnaire, Binge Eating Scale, and the Rosenberg Self-Esteem Scale. RESULTS: Data analyses revealed no significant differences between the two BED subgroups on any of the measures. DISCUSSION: These findings indicate that a history of purging behavior in BED is not associated with increased rates of comorbid psychopathology, severity of eating problems, dietary restraint, or attitudinal disturbance. Purging history does not appear to be a clinically meaningful variable with which to subclassify individuals with BED. PMID- 9741036 TI - Do unsuccessful dieters intentionally underreport food intake? AB - OBJECTIVE: A bogus pipeline paradigm was utilized to assess whether food intake underreporting by unsuccessful dieters is intentional. METHOD: Twenty-eight subjects completed 1-week food diaries. Then, 17 subjects in the experimental condition kept 2-week food diaries while being told the researcher was verifying their report. Eleven subjects in the control group were asked merely to self monitor for two more weeks. RESULTS: Results indicate that subjects in the experimental group reported significantly greater intake than control subjects, when controlling for reported intake during the screening phase and weight change. DISCUSSION: Thus, the belief that the researcher could verify their report improved the accuracy of patients' self-report. However, all subjects continued to underreport their dietary intake. In summary, underreporting may be an intentional attempt to manage presentation to others in a society that is increasingly critical of overweight persons. PMID- 9741037 TI - Heightened accuracy of self-reported weight in bulimia nervosa: a useful cognitive "distortion". AB - OBJECTIVE: This study was designed to test two competing hypotheses regarding bias in self-report of weight and height in bulimia nervosa. METHOD: General population samples of 102 young women with bulimia nervosa and 204 age and social class-matched healthy control women were recruited. Subjective and measured values of height and weight were obtained and compared within and between the groups. RESULTS: The healthy control subjects reported that they were lighter and taller than they actually were, and the degree of this discrepancy was related both to their actual weight and, more strongly, to the difference between their actual and desired weight. In contrast, the bulimia nervosa subjects showed little reporting bias. DISCUSSION: The results support the hypothesis that the intense interest of bulimia nervosa subjects in their weight is expressed by heightened precision of their self-report. Epidemiological studies of bulimia nervosa can capitalize on this cognitive "distortion." PMID- 9741038 TI - Measurement challenges and other practical concerns when studying massively obese individuals. AB - OBJECTIVE: To describe the measurement challenges faced and to evaluate the measurement quality obtained with massively obese individuals. METHOD: A cross sectional analysis of 107 individuals with body mass indices (kg/m2) > or = 50 was conducted. Individuals had their body fat measured via bioimpedance analysis (BIA), their serum leptin levels measured via radioimmunoassay (RIA), and height and weight measured via both laboratory scales and self-report. RESULTS: Serum leptin appeared to be measured accurately, provided the serum was diluted prior to conducting the RIA. Difficulties remained, however, in evaluating what was an unusual or expected value of leptin among individuals this large. Measures of impedance appeared to provide reasonable ordinal indications of body fatness. However, currently available equations for converting measures of impedance to estimates of percent body fat were highly inaccurate. Self-reported height and weight were reasonably good proxies of measured height and weight among individuals who reported their height and weight. However, a substantial proportion were unable to provide estimates. DISCUSSION: The above results suggest there are substantial challenges when trying to obtain meaningful measurements regarding obesity-related variables among massively obese individuals. Other logistic challenges also are discussed. It is hoped future research is directed at overcoming some of these challenges. PMID- 9741039 TI - Shape- and weight-based self-esteem and the eating disorders. AB - OBJECTIVES: To determine the psychometric properties of the Shape- and Weight Based Self-Esteem (SAWBS) Inventory in women with eating disorders, and to compare SAWBS scores in women who have eating disorders with women from psychiatric and normal control groups. METHOD: Women with eating disorders (n = 48), women with other psychiatric disorders (n = 44), and undergraduate control women (n = 82) completed the SAWBS Inventory and measures of depression, self esteem, and eating disorder symptomatology. Twenty women from the eating disorder group completed the SAWBS Inventory a second time 1 week later. RESULTS: Similar to previous work in undergraduate samples, SAWBS scores were stable over 1 week, and demonstrated concurrent and discriminant validity in women with eating disorders. In between-group comparisons, SAWBS scores were higher among women with eating disorders than in either control group, even after controlling for age, socioeconomic status, body mass index, and self-esteem. A differing relationship between depression and SAWBS emerged as a function of group; SAWBS scores differed significantly among depressed, but not nondepressed women from the three groups. CONCLUSION: The psychometric properties of the SAWBS Inventory were established in women with eating disorders. As expected, SAWBS scores were higher in women with eating disorders than in the control groups. Clinical implications of these findings are discussed. PMID- 9741040 TI - Body image and body weight in obese patients. AB - OBJECTIVE: To evaluate the influence of body weight on body image. METHODS: The study was carried out in severely obese patients and in postobese subjects, having attained and maintaining a normal or nearly normal weight following biliopancreatic diversion; body image was assessed by self-report questionnaires. RESULTS: The obese patients' scores were different from those of postobese subjects. In postobese individuals with adult-onset obesity, body image was very similar to that of controls, whilst in those with early-onset obesity it was abnormal. DISCUSSION: In the adult-onset obese patients, since the weight normalization causes a sharp improvement of body image, its alterations could be accounted for by a body shape far different from that socially acceptable. In the early-onset obese patients, being the postoperative findings similar to those of the obese patients and different from those of never-obese controls, the body image disparagement might reflect inner feelings, independent of body weight. PMID- 9741041 TI - Does the size of a binge matter? AB - OBJECTIVE: The aim of this study was to examine whether objective and subjective binges differ significantly from each other in relation to measures of psychopathology in a sample of women who meet DSM-IV diagnostic criteria for bulimia nervosa. METHOD: Baseline data from the Eating Disorder Examination (EDE) were analyzed and the average of the sum of and the difference between objective and subjective binge episodes were converted to z scores. Regressions were run with other baseline measures including the Structured Clinical Interview for Diagnosis of DSM-III-R (SCID) I and II disorders, EDE subscales, and psychological measures. RESULTS: We found no significant difference between the two types of binges on all but one measure, the "Can Do" subscale of the Self Efficacy Questionnaire, in a regression with the z score of total binges. DISCUSSION: The lack of significant findings questions the diagnostic validity of the "large amount of food" criterion used to define binge eating in the DSM-IV. PMID- 9741042 TI - Obese women with binge eating disorder define the term binge. AB - OBJECTIVE: The purpose of this study was to provide information regarding the criteria used by women with binge eating disorder (BED) to classify an eating episode a binge. METHOD: Sixty women who met DSM-IV research criteria for BED were interviewed and asked to define binge eating in their own words. Two independent raters classified subjects' responses according to a structured classification scheme. RESULTS: Loss of control over eating was the only criterion used to define binge eating by a majority (82%) of our subjects. Large amount of food and eating to relieve negative affect were reported less frequently, but appeared to be important criteria. DISCUSSION: The findings from this study are important to consider in an evaluation of the proposed DSM-IV research criteria for BED. PMID- 9741043 TI - Body image dissatisfaction and eating attitudes in visually impaired women. AB - OBJECTIVE: The high levels of body dissatisfaction and abnormal eating attitudes currently prevalent in Western societies have been attributed by many authors to the promotion of an unrealistically thin ideal for women. We investigated the role of the visual media by examining the relationship between body image dissatisfaction and eating attitudes in visually impaired women. METHOD: Questionnaires were administered to 60 women, 20 congenitally blind, 20 blinded later in life, and 20 sighted. RESULTS: Congenitally blind women had lower body dissatisfaction scores and more positive eating attitudes compared to women blinded later in life and sighted women, the latter having the highest body dissatisfaction scores and the most negative eating attitudes. Scores from sighted women were positively correlated with each other. DISCUSSION: The results suggest the importance of the visual media in promoting unrealistic images of thinness and beauty and are discussed from a sociocultural perspective. PMID- 9741044 TI - Very early-onset bulimia nervosa: report of two cases. AB - Bulimia nervosa is very rare in children below the age of 14 years, and no reliable reports of prepubertal bulimia nervosa have been published. We describe two cases of early-onset bulimia nervosa who presented before the age of 14 years, and with premenarchal onset in one patient. Both girls demonstrated high levels of the risk factors known to play a part in the etiology of bulimia nervosa. Implications of these cases regarding the etiology and occurrence of bulimia nervosa in younger adolescents are discussed. PMID- 9741045 TI - Disordered eating: a defense against psychosis? AB - The authors present four cases suffering with either bulimia nervosa or anorexia nervosa in conjunction with a psychotic illness. In all cases there appeared to be a reciprocal relationship between the eating disorder and psychosis such that improvement in eating precipitated or exacerbated the psychotic symptoms. We suggest that disordered eating serves as a defense against psychosis. Difficulties in treating such patients are discussed. PMID- 9741046 TI - Dexfenfluramine in psychotic patients. AB - Dexfenfluramine (DF) is contraindicated in severe psychiatric disorders and in depression. We used DF in 3 patients with chronic psychosis and severe overeating without changes in psychiatric pharmacotherapy. Two patients had paranoid schizophrenic psychosis with hallucinations, one patient mixed psychosis, beginning with lactation psychosis, and several attacks of hallucinations and depression later. Overeating was removed in all 3 patients without any negative effect on the psychotic state. All patients were able to maintain their body weight. Two patients with poorly controlled diabetes improved markedly their metabolic status. Doses up to 75 mg per day of DF were necessary during binge eating episodes in one patient. We conclude that DF can be used with care under close psychiatric supervision in psychotic patients with severe overeating. PMID- 9741047 TI - Rat testis mitochondrial aldehyde dehydrogenase: kinetic evidence for a direct reaction between capronaldehyde and enzyme-bound NAD+ in the presence of Mg2+ ions. AB - In the presence of Mg2+ saturation curves of aldehyde dehydrogenase show a sharp maximum at capronaldehyde concentrations lower than 1 microM. Since the native enzyme is a dimer, kinetic data have been analyzed with a general rate equation (given as a ratio of two polynomials) that takes into account the presence of two binding sites for both substrates and two for Mg2+. Simulation of the saturation curves was only successful after allowing the formation of the stable complexes ES, ES2, ES2M, ES2M2, EM and EM2. Since ESM and ESM2 are highly reactive but very unstable, activity at low aldehyde concentration can be explained by assuming a direct reaction mediated by Mg2+. At concentrations higher than 1 microM, capronaldehyde effectively binds to the enzyme in a highly cooperative process, but the formation of ES2M and ES2M2 results in slower reaction rates. Since ES2M2 is inactive, increase of the Mg2+ concentration eventually leads to strong inhibition. Experiments at different NAD+ concentrations show that the enzyme binds two NAD+, but reaction takes place at one binding site. PMID- 9741048 TI - Activation of transcription from the human immunodeficiency virus type 1 (HIV-1) long terminal repeat by the vasoactive intestinal peptide (VIP). AB - The ability of the human immunodeficiency virus type 1 (HIV-1) to persist and replicate in human CD4+ cells is under the control of both virally encoded proteins and a variety of host-related factors. Transcription of human immunodeficienty virus type 1 (HIV-1) is regulated by multiple cis-acting regulatory elements located in the viral long terminal repeats (LTRs). Here we report that the human vasoactive intestinal peptide (VIP) can activate the HIV-1 LTR. The activity of VIP is dose-dependent and specific. The effective concentration of VIP is within the physiological range suggesting that VIP release may modulate the activity of HIV-1 in vivo. A sequence in U3 region could be involved in VIP and in various VIP-related peptides' activation of HIV-1-LTR. This cis-acting element conferring VIP responsiveness is indistinguishable from HIV NF-kappa B tandem repeat binding sites (HIV-1 kappa B). PMID- 9741049 TI - Comparison of immune responses of two Salmonella gallinarum strains viewed as possible vaccines for fowl typhoid in Kenya. AB - The immune responses of two S. gallinarum strains, L46 and CN 180, were compared in 15-week-old cockerels. The humoral and cell-mediated immune responses were assayed by means of the indirect haemagglutination test (IHA) and the macrophage migration inhibition test (MIT), respectively. Birds were vaccinated with the two vaccines, respectively, and bled for sera (for IHA) and cells (for MIT) every week up to the seventh week, post vaccination, then every alternate week, three times, and later once every month, for a total period of 37 weeks. Strain L46 was found to induce an immune response that was very similar to that of CN 180. Both gave good humoral and cellular responses. PMID- 9741050 TI - Isolation and characterization of a Babesia species from Rhipicephalus evertsi evertsi ticks picked off a sable antelope (Hippotragus niger) which died of acute babesiosis. AB - Transmission of a Babesia species to susceptible cattle by Rhipicephalus evertsi evertsi ticks picked off a sable which died of acute babesiosis is described. The parasite was initially isolated by feeding R. e. evertsi ticks on a susceptible, splenectomised bovine which developed parasitaemia. Blood stabilate from the parasitaemic bovine produced a fatal babesiosis in a spleen--intact bovine. Clinical signs shown by the affected animals corresponded with those of acute babesiosis. Parasitological examination, the immunofluorescence antibody test and the polymerase chain reaction test revealed that the parasite transmitted by the ticks initially and the blood stabilates prepared from affected animals was Babesia bigemina. This parasite was morphologically identical to that observed in Giemsa-stained blood smears prepared from the dead sable. PMID- 9741051 TI - Observations on the use of Anaplasma centrale for immunization of cattle against anaplasmosis in Zimbabwe. AB - A total of 93 Bos taurus cattle was used in pen trials to compare vaccine stocks of Anaplasma centrale from South Africa and Australia (which stock came from South Africa in 1934) in protecting against three virulent field isolates from clinical Anaplasma marginale infections. In addition, field observations were made on the use of a vaccine, prepared from the Australian stock, in over 9553 cattle of mixed age and breeds on 16 co-operator farms and at one communal dip. The results of the pen trials indicated that the two vaccine stocks were comparable and that neither provided adequate protection against two of the three isolates of A. marginale. The field observations indicated that the vaccine was highly infective and produced mild reactions in most recipient cattle, and that users were generally satisfied with the vaccine. These somewhat conflicting results are discussed in the context of observations in Australia and future vaccination against anaplasmosis in Zimbabwe. PMID- 9741052 TI - Three new species of ciliated Protozoa from the hindgut of both white and black wild African rhinoceroses. AB - This report deals with the effect of the mode of feeding of the hindgut fermenting herbivorous rhinoceros on the species of Protozoa fermenting the ingesta, as demonstrated by the proposed three new species of ciliated Protozoa: Didesmis synciliata differing from D. ovalis in having syncilia in place of simple cilia, Blepharoconus dicerotos being twice the size of B. cervicalis, and Blepharosphaera ceratotherii being one third the size of B. intestinalis. The findings are in line with the biological tenet that in herbivores the composition of the diet is the major factor determining the composition of the digestive organisms. PMID- 9741053 TI - Immunization of rabbits with Amblyomma hebraeum nymphal homogenates and implications for the host amplification system. AB - Immunochemical mechanisms involved in tick rejection by a host are not well documented. The role of serum globulins, and that played by the amplification system's humoral products (thrombin from the coagulation, plasmin from the fibrinolytic, and kallikrein from the kinin systems) in tick-resistant animal hosts have not yet been demonstrated. It is known, however, that factors C1, C3 and C5 of the complement system play a role in tick rejection, and that factors C3a and C5a are anaphylatoxins capable of degranulating leukocytes, thereby releasing pharmacologically active vasoamines which are involved in tick rejection. In this study, levels of kininogen increased by 56% and those of fibrinogen by 19% in rabbits immunized with nymphal antigens. A highly significant (P < 0.001) number of nymphs that fed on the immunized rabbits failed to moult into adult stages. It is reported for the first time, that increased levels of two glycoproteins, fibrinogen and kininogen occurred in rabbits immunized with homogenates of Amblyomma hebraeum ticks. The role played by the amplification system in tick rejection in resistant animals is clarified. PMID- 9741054 TI - The clinical efficacy of enrofloxacin in the treatment of experimental bovine pneumonic pasteurellosis. AB - The clinical efficacy of enrofloxacin was tested in calves with experimentally induced pneumonic pasteurellosis. A strain of Pasteurella haemolytica, biotype A, serotype 1 (P. haemolytica A1), which had been isolated from an outbreak of pneumonic pasteurellosis in feedlot calves, was used to induce the disease in 24 eight-month-old calves. Each animal received, by intratracheal injection, 6 x 10(11) colony forming units of P. haemolytica A1 in a four-hour log phase culture. Twelve similar animals were kept as non-infected controls (Negative Control group). Treatment of the infected animals commenced 40 h after infection and was as follows: 12 animals each received 2.5 mg/kg enrofloxacin subcutaneously and 12 animals each received 5 ml sterile saline intramuscularly (Positive Control group). All treatments were given once daily for three consecutive days. Clinical examinations were performed on all animals once daily, starting prior to infection and continuing until 12 d post-infection. The parameters evaluated were rectal temperature, habitus (attitude), ocular mucous membrane congestion and abnormal sounds on lung auscultation. On day 14 post infection, all animals were killed and their lung lesions (if any) estimated as the percentage involvement of each pair of lungs. The only statistically significant (P > or = 0.05) differences observed were between the Negative Control group and the Positive Control group. Noticeable differences were seen between the enrofloxacin-treated group and the Positive Control group, but they were not statistically significant (P > 0.05). The average lung lesion score (pneumonic lesions as a percentage of total lung volume) for the Positive Control group was 12.1% and that of the enrofloxacin-treated group, 8.4%. This difference was not statistically significant (P > 0.05). PMID- 9741055 TI - Ixodid tick infestations of wild birds and mammals on a game ranch in central province, Zambia. AB - Ticks were collected at irregular intervals from December 1995 to November 1996 from wildlife on Mtendere Game Ranch in the Chisamba District of Central Province, Zambia. Total collections were made from two species of ground-nesting birds and 20 species of small and large mammals. Thirteen species/subspecies of ixodid ticks were recovered. Rhipicephalus appendiculatus was the most abundant, followed by Boophilus decoloratus and Rhipicephalus evertsi evertsi. Small numbers of immature ticks of only a few species were collected from the birds and rodents. The lagomorphs carried large numbers of predominantly immature R. appendiculatus. Most of the ungulates harboured several tick species and had high infestations of R. appendiculatus. The seasonal abundances of Amblyomma variegatum, B. decoloratus, R. appendiculatus and R. evertsi evertsi were determined. PMID- 9741057 TI - An epidemic of besnoitiosis in cattle in Kenya. AB - A total of 17 head of cattle presenting with typical "Elephant skin disease" were isolated from the rest of the herd within a beef ranch for further clinical observation. On physical examination, all the animals had characteristic sclero conjunctival cysts of Besnoitia besnoiti. In addition, some of the animals had characteristic skin nodules on the legs, ears and back. Histological examination of skin sections revealed typical large Besnoitia cysts. Microscopic examination of crush preparations of skin scrapings revealed crescent-shaped organisms with a more pointed anterior than posterior end (banana-shaped morphology) confirming that the cysts belong to the genus Besnoitia. PMID- 9741056 TI - Selection of an scFv phage antibody that recognizes bluetongue virus from a large synthetic library and its use in ELISAs to detect viral antigen and antibodies. AB - A filamentous phage library displaying a vast repertoire of synthetic single chain fragment variable (scFv) antibody fragments was subjected to affinity selection on purified bluetongue virus (BTV) particles. After four rounds of selection and amplification, 73 out of a total of 90 fusion phage clones tested were found to bind to purified BTV in ELISA. One of these, the clone producing the highest ELISA signal, was selected for an investigation of its potential as an immunodiagnostic reagent. The binding of this phage antibody (designated A12) could be inhibited by free virus and by antibodies in immune serum. Inhibition with antibodies in guinea-pig sera suggested that it recognized an antigenic region on BTV that was similar on at least 10 different BTV serotypes. A sandwich ELISA utilizing antibody A12 was capable of detecting approximately 60 ng of purified BTV. PMID- 9741058 TI - Helminths and bot fly larvae of wild ungulates on a game ranch in central province, Zambia. AB - Helminths and bot fly larvae were collected from 11 wild ungulate species on a game ranch in the Central Province of Zambia. New host-parasite records are: Calicophoron sp. from defassa waterbuck Kobus ellipsiprymnus defassa and Kafue lechwe Kobus leche kafuensis; Avitellina centripunctata, Gaigeria pachyscelis and Gedoelstia cristata from tsessebe Damaliscus lunatus lunatus; Cooperia rotundispiculum from common reedbuck Redunca arundinum; Dictyocaulus filaria from greater kudu Tragelaphus strepsiceros; Dictyocaulus sp. from tsessebe and defassa waterbuck and Strobiloestrus sp. from sable antelope Hippotragus niger. Most of the other parasites collected are first records for Zambia and thus extend the distribution ranges of several species. PMID- 9741059 TI - A system for the brain-enhanced delivery of estradiol: an assessment of its potential for the treatment of Alzheimer's disease and stroke. PMID- 9741060 TI - Synthesis and antiviral activity of 7-O-(omega-substituted)-alkyl-3-O methylquercetin derivatives. AB - 3',4'-Di-O-benzyl-3-O-methylquercetin (2), the precursor in the synthesis of an important antivirally active flavone 3-O-methylquercetin (1), was regioselectively alkylated at the 7-OH position by a series of 1,omega dihaloalkanes and omega-bromoalkanols. Dimerization of the flavone had to be avoided by applying strict reaction conditions. Subsequent debenzylation was carried out by catalytic transfer hydrogenolysis, affording quantitatively the 7 O-(omega-haloalkyl)-3-O-methylquercetin (11-14) and 7-O-(omega-hydroxyalkyl)-3-O methylquercetin derivatives (15, 16). All compounds were tested for their antiviral activity against poliomyelitis- and HIV-viruses. PMID- 9741061 TI - Theophylline derivatives as potential histamine H3-receptor antagonists. AB - Previous results of histamine H3-receptors investigations allowed to formulate a general structure of H3-receptor antagonists. According to this model a series of compounds were obtained. As heterocycles they contained a theophylline moiety connected with a polar group (amine, ester, amide, and thiourea function) via an alkyl chain linked by a spacer to a lipophilic residue. The common distance between xanthine moiety and lipophilic rest was a six-link-chain. Selected compounds did not show significant H3-receptor antagonist activity and were weak antagonists at histamine H1-receptors. PMID- 9741062 TI - Non-steroidal anti-inflammatory agents: synthesis of novel benzopyrazolyl, benzoxazolyl and quinazolinyl derivatives of 4(3 H)-quinazolinones. AB - Four novel series of 4(3 H)-quinazolinone derivatives have been synthesized by cyclization of the intermediate 3-aryl-2-(6-aryl-2-cyclohexen-1-on-5-yl)-4(3 H) quinazolinones 3a-f with hydrazine, phenylhydrazine, hydroxylamine and thiourea. The products are 3-aryl-2-(6-aryl-3-methyl-1 H-4,5-dihydrobenzo[d]pyrazol-4-yl) 4(3 H)-quinazolinones 4a-f; 3-aryl-2-(6-aryl-3-methyl-1-phenyl-1 H-4,5 dihydrobenzo[d]pyrazol-4-yl)-4(3 H)-quinazolinones 4g-1; 3-aryl-2-(6-aryl-3 methyl-4,5-dihydrobenzo[d]-1,2-oxazol-4-yl)-4(3 H)-quinazolinones 5a-f, and 3 aryl-2-(7-aryl-4-methyl-5,6-dihydro-2(1 H)thioxoquinazolin-5-yl)-4(3 H) quinazolinones 6a-f. Some of these compounds showed anti-inflammatory activity comparable to or higher than that of the reference compound proquazone. PMID- 9741063 TI - Pyridines and pyrazolines from salicylic acid derivatives with propenone residue and their antimicrobial properties. AB - Reaction of the propenones 1c, d with chlorosulfonyl isocyanate followed by hydrolysis gave the corresponding carbamoyloxybenzoates 2a, b. While their reaction with ethyl isocyanate afforded the 1,3-benzoxazine-2,4-diones 3a, b. Reaction of 1a, b with aryl hydrazines gave the pyrazolines 4a, d, whereas, with hydrazine hydrate in acetic acid, the acetyl derivatives 4e, f were produced. 1c, d reacted with malononitrile and ethyl cyanoacetate affording the cyanopyridines 5 and cyanopyridones 6 respectively. The products show antimicrobial activities. PMID- 9741064 TI - Cyclodextrins in acetazolamide eye drop formulations. AB - The interaction of acetazolamide with beta-cyclodextrin, (beta-CD), dimethyl-beta cyclodextrin (DM-beta-CD) and trimethyl-beta-cyclodextrin (TM-beta-CD) was monitored spectrophotometrically. The results revealed formation of equimolar complexes. The apparent solubility of acetazolamide in water was found to increase linearly with increasing CD concentration. The effect of CDs on the permeation of acetazolamide through semi-permeable membranes and the topical delivery of acetazolamide was investigated. Maximum acetazolamide penetration was obtained when just enough CD was used to keep all acetazolamide in solution. For an acetazolamide concentration of 10 mg/ml, the optimum CD concentration appeared to be 3.5 mmol/l for beta-CD, 2.8 mmol/l for TM-beta-CD and 6.0 mmol/l for DM beta-CD. The effect of CDs on the bioavailability of acetazolamide was assessed by measuring the intraocular pressure in rabbits. The results indicated that CDs have a significant influence on the biological performance of the drug leading to augmentation in its intensity of action and bioavailability as well as prolongation in its duration of action. PMID- 9741065 TI - Inhibition of corticotropin releasing factor (CRF)-induced adrenocorticotropin (ACTH) secretion in pituitary corticotropic cells by substance P. AB - Substance P (SP) is one of the three distinct peptides of tachykinin system which possess a common spectrum of biological activities including a modulation of stress. It is assumed that the anterior pituitary is one possible target of SP in attenuation the stress response. Therefore the interaction between the hypothalamic stress hormone corticotropin releasing factor (CRF) and SP was investigated in AtT20/D16v-cells, a cellular model derived from a pituitary tumor. CRF stimulates the release of ACTH from AtT20/D16v cells in a concentration dependent manner. SP (1 microM) was able to abolish the CRF (100 nM)-induced ACTH release. In the same way SP inhibited the CRF-induced accumulation of cyclic adenosine monophosphate (cAMP), indicating that SP influenced the signal transduction pathway of CRF receptor activation. Thus, a direct inhibition of the CRF-mediated stress response by SP at the level of anterior pituitary seems to be likely. PMID- 9741066 TI - Corollosporine, a new phthalide derivative from the marine fungus Corollospora maritima Werderm. 1069. AB - Extracts of the culture medium from the marine fungus Corollospora maritima exhibited concentration dependent antibacterial activity against Staphylococcus aureus and other microorganisms. Bioactivity-guided fractionation and purification afforded the new isobenzofuranone or phthalid type compound corollosporine. PMID- 9741067 TI - Concurrent psychotherapy and pharmacotherapy in the treatment of depression and anxiety in cancer patients. AB - Over the past 30 years, numerous studies have compared the relative effectiveness of psychotherapy, pharmacotherapy, and concurrent therapeutic approaches in treatment of common psychiatric disorders, such as depression and anxiety. Generally, these studies have demonstrated that the combined approach is somewhat more effective in treating the disorder in question, as well as in preventing relapse. A number of theories regarding the reasons for this finding have been proposed. The application of this therapeutic approach to the specific population of cancer patients has not been studied in systematic research. This paper briefly reviews the studies comparing the efficacies of various treatment approaches for depression, discusses reasons for the efficacy of such an approach, and outlines a series of reasons why this may be the preferred model for intervening with cancer patients. PMID- 9741068 TI - A controlled trial of fluoxetine and desipramine in depressed women with advanced cancer. AB - BACKGROUND: This study was conducted to determine the efficacy and tolerability of fluoxetine and desipramine in treating depressive symptoms in women with cancer. METHOD: In this prospective, 6-week, double-blind, placebo-controlled trial, we compared fluoxetine with desipramine in treating depressive symptoms in 40 women diagnosed with cancer. Scales used to measure efficacy and tolerability were the Hamilton Depression Rating Scale (HAM-D), the Hamilton Anxiety Rating Scale (HAM-A), the Clinical and Patient's Global Impression (CGI and PGI) scales, the Functional Living Index for Cancer (FLIC), the Memorial Pain Assessment Card (MPAC), and the SF-36 Health Survey. RESULTS: Fluoxetine and desipramine treatments improved depression and anxiety symptoms. There was a trend towards significance in improvement of FLIC scores (as evidenced by greater numerical improvements with fluoxetine treatment). Fluoxetine treatment alone was associated with statistically significant improvements in MPAC Mood scale scores. Both treatments showed statistically significant improvements in the quality of life SF-36 scores in Role Emotional, Social Functioning, Mental Health, and Vitality. CONCLUSIONS: Both fluoxetine and desipramine were effective and well tolerated in improving depressive symptoms and quality of life in women with advanced cancer. Fluoxetine may offer greater benefit to these patients, as evidenced by greater improvements in fluoxetine-treated patients on several quality of life measures. Our results, while meaningful, should be confirmed in a larger patients sample. However, experience from studies of antidepressant use in patients with advanced cancer has shown that intercurrent disease and treatment variables make it difficult to conduct large studies. PMID- 9741069 TI - The use of psychotropic medication in patients referred to a psycho-oncology service. AB - The purpose of this study was to investigate the use of psychotropic medication in patients referred to a psycho-oncology service. While depressive disorders and psychological difficulties are being increasingly recognised in oncology patients, the use of psychotropic medication has not been frequently studied, nor has it been studied in patients who are subsequently referred for psychiatric assessment. The use of psychotropic medication in all patients referred to a psycho-oncology service over a 6-month period was examined prospectively. Details recorded included class of psychotropic medication used and by whom it was prescribed. Demographic details and clinical diagnoses were also recorded. Sixty three patients were referred over the initial 6 months of the service and the majority had advanced or metastatic disease (62%). Clinically, 44.5% had some form of major psychiatric disorder, and 40% had an adjustment disorder. Over half (55.5%) were already on psychotropic medication at referral; mainly minor tranquillisers (51%) and antidepressants (24%); 22% were on more than one drug. Of those medications prescribed pre-referral, 46% had been prescribed by the oncology team and 31% were from the GP. Following psychiatric review, further medication was prescribed in 30% of the subjects, leaving a total of 79% on some form of psychotropic drug. While the overall psychotropic prescribing and the use of minor tranquillisers appears to be similar to those found in earlier studies, the high rate of use of antidepressants suggests psychological distress is being increasingly recognised, and pharmacotherapy is a commonly used strategy in this group. PMID- 9741070 TI - Psychotropic drug metabolism in the cancer patient: clinical aspects of management of potential drug interactions. AB - An overview of drug metabolism, with particular focus on the cytochrome p450 system, is provided in this review. To date, there has been a growing body of literature concerning the cytochrome p450 enzyme, drug-drug interactions and the role of psychotropic medications when co-administered with medications prescribed in the medically ill population. The article provides an ability to cross reference commonly prescribed medications to their known involvement as substrates, inhibitors or inducers of p450 enzymes. This information will permit the clinician working in an oncologic setting to better predict potential interactions based on available in vitro and in vivo data and choose psychotropics analytically when confronted with a situation of polypharmacy. A knowledge of drug interactions will decrease the uncertainty in prescription of multidrug therapies and minimize the likelihood of diminished drug efficacy or toxic reactions. PMID- 9741071 TI - Using computer databases to predict and avoid drug-drug interactions in the cancer patient requiring psychotropics. AB - Psychotropic drug interactions with medical prescriptions is an essential knowledge base for the practicing oncologist as well as the psychiatrist. This paper describes a method to identify, understand, and develop a conceptual framework to codify psychotropic-medical drug interactions; a systemic approach for updating this key database, and a software system to assist in the management of this ever unfolding sphere of knowledge; and, a set of instruments to provide ongoing developments in this important area. Since many of the trials either for oncological medications or psychotropic drugs did not include studies comparing these two domains, their interactions often only become known with their utilization and subsequent reporting. PMID- 9741072 TI - Psychotropic adjuvant analgesics for pain in cancer and AIDS. AB - The 'WHO Analgesic Ladder' is a well validated approach for the selection of appropriate analgesic therapy for cancer pain as well as pain in AIDS. The mainstay of analgesic intervention for cancer and AIDS pain of moderate to severe intensity continues to be the appropriate use of opioid analgesics. There is, however, a growing appreciation for the role of adjuvant analgesics, such as antidepressants and other psychotropic medications, at each step of the WHO Analgesic Ladder, particularly in the treatment of neuropathic pain. Knowledge of the indications and usefulness of psychotropic analgesic drugs in cancer and AIDS pain populations will be most important to clinicians practicing in psycho oncology/AIDS settings, particularly since these drugs are useful not only in the treatment of psychiatric complications of cancer and AIDS, but also as adjuvant analgesic agents in the management of pain. This paper reviews the literature on the use of antidepressants, psychostimulants, neuroleptics, anticonvulsants and other psychotropic analgesics in the management of cancer and AIDS pain. Mechanisms of analgesia, drug selection, and recommendations for clinical usage are discussed. The appropriate and timely use of psychotropic adjuvant analgesic drugs represents an opportunity for active psychiatric contribution to the multidisciplinary management of cancer and AIDS pain. PMID- 9741073 TI - The uses of psychotropics in symptom management in advanced cancer. AB - Approximately 50% of patients diagnosed with cancer die because of progressive disease. Psychotropic drugs are frequently used for the management of physical and psychosocial symptoms in these patients. Thalidomide, cannabinoids and melatonin are emerging agents for the management of cachexia. Psychostimulants have a defined role in the management of opioid-induced sedation. Haloperidol, tricyclic anti-depressants and newer anti-depressants also have an established role in the management of neuropsychiatric symptoms such as delirium or depression. Cancer patients present unique challenges for successful psychotropic therapy including older age, malnutrition, autonomic failure, borderline cognition, opioid and psychotropic therapy. A practical clinical approach which defines a specific target symptom, an outcome latency period, expected side effects, and reviews possible drug interactions, and frequent monitoring is outlined. Continued research is needed to further define the role of psychotropics in the management of the different physical and psychosocial symptoms in advanced cancer patients. PMID- 9741074 TI - Conformational study of synthetic delta 4-uronate monosaccharides and glycosaminoglycan-derived disaccharides. AB - Sixteen delta 4-uronate monosaccharides were chemically synthesized. Their carboxy group was protected as a methyl or benzyl ester, the anomeric hydroxyl group as a benzyl glycoside and the 2 and 3 hydroxyl groups were protected with different substitution patterns as both ester and ether derivatives. Disaccharides containing delta 4-uronates were prepared from heparin layses. Their carboxy group was unprotected or protected as a benzyl ester and the two hydroxyls in the uronate moiety were free, as O-sulfo derivates or acylated. The conformation of these unsaturated uronate monosaccharide and disaccharide residues was studied using 1H NMR by examining interproton vicinal coupling constants. The delta 4-uronate residue adopted either the 2H1 or the 1H2 conformations. The equilibrium between these two conformers was shown to be controlled by substitution pattern. PMID- 9741075 TI - Synthesis of hyaluronic-acid-related oligosaccharides and analogues, as their 4 methoxyphenyl glycosides, having N-acetyl-beta-D-glucosamine at the reducing end. AB - To contribute to the possibilities to study the ability of oligosaccharide fragments of hyaluronic acid to induce angiogenesis, several hyaluronic-acid related oligosaccharides and their 6-O-sulfated analogues were synthesised as their 4-methoxyphenyl glycosides having 2-acetamido-2-deoxy-D-glucopyranose at the reducing end. In all syntheses described, the D-glucopyranosyluronic acid residue was obtained by oxidation at C-6 of a corresponding D-glucopyranosyl residue after construction of the oligosaccharide backbone, using pyridinium dichromate and acetic anhydride. PMID- 9741076 TI - Preparation of spacer-containing di-, tri-, and tetrasaccharide fragments of the circulating anodic antigen of Schistosoma mansoni for diagnostic purposes. AB - The chemical synthesis of beta-D-GlcpA-(1-->3)-beta-D-GalpNAc-(1-->O)CH2CH = CH2, beta-D-Galp-NAc-(1-->6)-[beta-D-GlcpA-(1-->3)]-beta-D-GalpNAc-(1-- >O)CH2CH = CH2, and beta-D-GlcpA-(1-->3)-beta-D-GalpNAc-(1-->6)-[beta-D-GlcpA-(1 -->3)] beta-D-GalpNAc-(1-->O)CH2CH = CH2 is described. These oligosaccharides represent fragments of th circulating anodic antigen, secreted by the parasite Schistosoma mansoni in the circulatory system of the host. The applied synthesis strategy includes the preparation of a non-oxidised backbone oligosaccharide, with a levulinoyl group at O-6 of the beta-D-glucose residue. After the selective removal of the levulinoyl group, the obtained hydroxyl functions were converted into carboxyl groups, using pyridinium dichromate and acetic anhydride in dichloromethane, to afford the desired glucuronic-acid-containing oligosaccharides. Subsequently, the allyl glycosides have been elongated with cysteamine to give the corresponding amine-spacer-containing oligosaccharides. PMID- 9741077 TI - Synthesis of a tri- and tetradeoxy analogue of methyl 3,6-di-O-alpha-D mannopyranosyl-alpha-D-mannopyranoside for investigation of the binding site of various plant lectins. AB - The synthesis of a the 2,4,3'-trideoxy and 2,4,3',4'-tetradeoxy analogues of the trimannoside part of the core structure of N-linked glycoproteins, methyl 3,6-di O-alpha-D-mannopyranosyl-alpha-D-mannopyranoside, is described. A 2,4-dideoxy (1- >6) linked disaccharide was used as a common intermediate acceptor, which was coupled with a 3-deoxy and a 3,4-dideoxy benzochlorosugar donor, the latter prepared from methyl alpha-D-mannopyranoside in five steps. Despite the acid sensitive donors and acceptor, acceptable glycosylation yields were obtained of both the trideoxy- and the tetradeoxy trisaccharide using silver triflate as a promoter (65 and 51%, respectively). Deprotection in one step then gave the target products. PMID- 9741078 TI - Construction of insertion and deletion mxa mutants of Methylobacterium extorquens AM1 by electroporation. AB - Methylobacterium extorquens AM1 is a pink-pigmented facultative methylotroph which is widely used for analyzing pathways of C1 metabolism with biochemical and molecular biological techniques. To facilitate this approach, we have applied a new method to construct insertion or disruption mutants with drug resistance genes by electroporation. By using this method, mutants were obtained in four genes present in the mxa methylotrophy gene cluster for which the functions were unknown, mxaR, mxaS, mxaC and mxaD. These mutants were unable to grow on methanol except the mutant of mxaD, which showed reduced growth on methanol. PMID- 9741079 TI - Analysis of sisomicin binding sites in Micromonospora inyoensis cell wall. AB - Sisomicin binding sites are located in the cell wall. They are the carboxyl groups of peptidoglycans, which are major components of the cell wall. The carboxyl groups have negative charges which can bind the positively charged amino groups of sisomicin. When the negative charges of the carboxyl groups of the peptidoglycans are changed to neutral or positive charges, sisomicin does not bind to the cell wall. Magnesium ions bind to the cell wall in competition with sisomicin, and have a weak affinity for the cell wall binding sites compared with sisomicin. The binding molar ratio of sisomicin to magnesium ions was approximately 1 to 10. PMID- 9741080 TI - Two methods for the genetic differentiation of Lactococcus lactis ssp. lactis and cremoris based on differences in the 16S rRNA gene sequence. AB - The 16S ribosomal RNA gene sequences of Lactococcus lactis ssp. lactis and ssp. cremoris differ by 9-10 bp (depending on strain), within the first 200 bp of the sequence. These differences were used to develop two methods of genetically differentiating lactis and cremoris strains. Primers to conserved sequences in the 16S rRNA gene were used in a PCR reaction to amplify fragments of the 16S rRNA gene. A single base difference at position 180 of the sequence was utilised to develop a ligase chain reaction to differentiate lactis and cremoris sequences. The second method involved digestion of the amplified fragments with restriction endonucleases specific for either the lactis or cremoris sequence. Resolution of the digested fragments on an agarose gel allowed the strains to be identified as genetically lactis or cremoris. This method was used to examine lactococci isolated from raw milk. Of 31 raw milk strains examined, 21 contained the cremoris 16S rRNA sequence, however, all 31 strains exhibited the phenotypic characteristics of the lactis subspecies. PMID- 9741081 TI - PCR amplification and characterization of the intergenic spacer region of the ribosomal DNA in Pyrenophora graminea. AB - Successful amplification of the whole intergenic spacer region of the nuclear ribosomal repeat (IGS) in Pyrenophora graminea was obtained with a PCR-based assay. Single amplification products showed length differences. Depending on the length of the IGS-PCR product, ca. 3.8 or 4.4 kb, two groups of isolates could be identified. The RFLP patterns of isolates obtained with the 6-base cutting enzymes Apal, BglII, DraI, EcoRV, HindIII and SacI were similar within each group and different between the two groups. Restriction patterns of IGS-PCR products digested with the 4-base cutting enzyme AluI were polymorphic among isolates in spite of their IGS-PCR product length. In order to characterize the long and short IGS-PCR products the restriction map is shown. The long product shows an additional HindIII site and a BglII site that is lacking in the short product. However, the latter shows a SacI site that is not present in the long IGS-PCR product. Therefore, the described PCR-RFLP analysis of the IGS appears to be a useful tool to resolve genetic variation between P. graminea isolates. PMID- 9741082 TI - A two-component system involved in regulation of anaerobic toluene metabolism in Thauera aromatica. AB - The genes for a two-component regulatory system of the denitrifying toluene degrading bacterium Thauera aromatica were identified immediately upstream of the genes for benzylsuccinate synthase (bssDCAB), the first enzyme involved in anaerobic toluene metabolism. The genes apparently encode the regulators of toluene catabolic enzymes and were therefore termed tdiSR (for toluene degradation including sensor and regulator). The tdiR gene product was overproduced in Escherichia coli and assayed for binding to a DNA fragment containing the 5' region of the bss operon. We observed specific DNA binding with cell extracts containing overproduced TdiR, but not with control extracts. The tdiSR genes are almost identical to two genes of Thauera strain T1, which have not been assigned a function so far. In addition, the derived gene products share similarity with regulators of toluene and styrene catabolic pathways in aerobic Pseudomonas species, and with the tutCB gene products of Thauera strain T1. The latter have previously been implicated in regulating anaerobic toluene metabolism. Our data suggest that toluene catabolism under aerobic and anaerobic conditions is regulated by similar, but distinct two-component systems. PMID- 9741083 TI - Isolation of an Escherichia coli O157:H7 strain producing Shiga toxin 1 but not Shiga toxin 2 from a patient with hemolytic uremic syndrome in Korea. AB - Escherichia coli strains isolated from patients with diarrhea or hemolytic uremic syndrome (HUS) at Pusan University Hospital, South Korea, between 1990 and 1996 were examined for traits of the O157:H7 serogroup. One strain isolated from a patient with HUS belonged to the O157:H7 serotype, possessed a 60-MDa plasmid, the eae gene, and ability to produce Shiga toxin 1 but not Shiga toxin 2. Arbitrarily primed PCR analysis suggested that this strain is genetically very close to a O157:H7 strain isolated in Japan. PMID- 9741084 TI - The O7 antigen of Stenotrophomonas maltophilia is a linear D-rhamnan with a trisaccharide repeating unit that is also present in polymers for some Pseudomonas and Burkholderia species. AB - The O antigen polymer recovered from the reference strain for Stenotrophomonas (Xanthomonas or Pseudomonas) maltophilia serogroup O7, after mild acid hydrolysis of the lipopolysaccharide, was constructed from D-rhamnose. By means of chemical degradations and NMR studies, the repeating unit of the polymer was shown to be a linear trisaccharide with the structure -->2)-alpha-D-Rhap-(1-->3)-alpha-D-Rhap (1-->3)-alpha-D-R hap-(1-->. The same repeating unit is present in the common antigen of Pseudomonas aeruginosa and in O antigens from some pathovars of Pseudomonas syringae and a strain of Burkholderia (Pseudomonas) cepacia. PMID- 9741085 TI - A novel proline-rich protein, EspF, is secreted from enteropathogenic Escherichia coli via the type III export pathway. AB - Enteropathogenic Escherichia coli (EPEC) cause a characteristic attaching and effacing (A/E) lesion in intestinal epithelial cells that is associated with the expression and export of specific bacterial proteins via a type III secretion pathway. These effector proteins and components of the type III export apparatus are encoded on a pathogenicity island known as the locus of enterocyte effacement (LEE). In this study, we describe a proline-rich protein, EspF, encoded by the LEE that is secreted by the EPEC type III secretion apparatus. Whereas an espF deletion mutant does not synthesize or secrete EspF, surprisingly it retains the ability to induce host signaling events, perform A/E activities, and invade host epithelial cells. Although these results do not indicate on obvious role for EspF in the formation of A/E lesions nor in the invasion of epithelial cells, they do not preclude a role played by EspF in other aspects of EPEC pathogenesis. PMID- 9741086 TI - Serotype conversion of a Shigella flexneri candidate vaccine strain via a novel site-specific chromosome-integration system. AB - Shigella flexneri SFL124 (serotype Y) is a promising live oral vaccine candidate, which has been shown to be safe and immunogenic in human volunteers. To change the serotype of this vaccine strain, we inserted a serotype conversion gene cluster into the chromosome of SFL124 by using a bacteriophage-based site specific integration system. By cloning an integrase gene (int), an attachment site (attP) and a glucosyl transfer gene cluster from bacteriophage SfX into a suicide vector, and subsequently introducing this construct into S. flexneri SFL124, we obtained a S. flexneri strain (designated SFL1213) expressing the serotype X somatic antigen specificity. The strain retained other characteristics of the parent strain, such as colony shape, growth rate, and Congo red binding property. Stability test showed that the serotype X O-antigen specificity in SFL1213 was 100% stable after being cultured approximately 72 successive hours under non-selective condition. In a mouse pulmonary model, the recombinant strain elicited a significant level of humoral antibodies which recognized the lipopolysaccharide (LPS) of a wild-type S. flexneri serotype X strain. The site specific insertion system will be useful when stable expression of a cloned single copy gene is desired in the chromosome of S. flexneri vaccine candidate, SFL124. PMID- 9741087 TI - Unique morphogenesis in Saccharomyces cerevisiae strain GS1731. AB - During the lag and early exponential phase of growth, 50-60% of budded cells of Saccharomyces cerevisiae strain GS1731 were multiply budded. During subsequent culture growth, the frequency of multiply budded cells decreased until by stationary phase multiply budded cells were rare. Data from renewed growth of a culture after hydroxyurea treatment indicated that GS1731 mother cells could assemble up to three pre-bud sites and begin bud growth and development in each. Light and scanning electron microscopy showed two or three very small buds emerging simultaneously on a mother cell and either reaching full size at the same time or enlarging sequentially. Immunofluorescence studies revealed that these multiply budded cells had multiple bundles of cytoplasmic microtubules. DAPI staining of nuclei revealed that some of the unbudded mother cells were multinucleate and completed cytokinesis giving rise to normal daughter cells. PMID- 9741088 TI - Conditions that induce Staphylococcus aureus heat shock proteins also inhibit autolysis. AB - When Staphylococcus aureus strain 8325 was grown at 30 degrees C and heat shocked at 40 degrees C the rate of cell autolysis in buffer with or without Triton X-100 was reduced. Treatment of growing cells with other agents (CdCl2, ethanol, NaCl) known to induce heat shock proteins also resulted in cells that showed a decreased rate of autolysis. Heat shocked cells showed lower rates of freeze-thaw autolysin activity on purified cell walls, and isolated crude cell walls from heat shocked cells had lower rates of autolytic activity compared to controls. No differences in the peptidoglycan hydrolase activity profiles of control and heat shocked cells were detected by renaturing sodium dodecyl sulfate polyacrylamide gel electrophoresis. It is proposed that autolysins are damaged by heat shock and their targeting to the cell wall is impaired, possibly by complexing with heat shock proteins, which may also inhibit autolysin activity. Heat shock also inhibited the autolytic activity of methicillin-resistant and related-susceptible strains, and the possible relationship of this to the expression of methicillin resistance is discussed. PMID- 9741089 TI - Identification of a gene sequence encoding a putative pyruvate oxidoreductase in Serpulina pilosicoli. AB - Serpulina pilosicoli is a recently described species of intestinal spirochaete which can be identified using a species-specific monoclonal antibody BJL/AC1 reactive with a 29-kDa protein located in the cell envelope. A genomic library of the type strain of S. pilosicoli P43/6/78T was created in lambda zap express and screened using BJL/AC1. Single positive clones were isolated and excised into the phagemid vector pBK-CMV. Phagemid DNA was purified and a single clone was selected for sequencing. The size of spirochaetal DNA insert was determined by digestion with restriction endonucleases EcoRI and PstI as being approximately 2.6 kb. The nucleotide sequence of the gene encoding the protein with which the antibody reacted was determined by cycle sequencing. The insert contained an open reading frame of 285 nucleotides. Translation of the nucleotide sequence into amino acid (aa) residues showed a sequence of 275 aa. Comparison of this sequence with databases revealed homology to pyruvate oxidoreductases from various organisms found in the gastroinestinal tract. These included the pyruvate ferredoxin oxidoreductase (POR) alpha submit of Helicobacter pylori (38.8% identity in 250 aa), pyruvate-flavodoxin oxidoreductase of Escherichia coli (28.7% identify in 258 aa) and Giardia intestinalis (25.1% identity in 251 aa). A significant level of homology was also observed with hyperthermophilic bacteria such as the POR of Thermatoga maritima (38.6% in 254 aa) and the 2-ketovalerate ferredoxin oxidoreductase of Pyrococcus furiosus (34% in 262 aa). PMID- 9741090 TI - Human transferrin as a source of iron for Streptococcus intermedius. AB - Streptococcus intermedius is well known to produce severe infections in various areas of the body. In this study, we evaluated the ability of S. intermedius to utilise human transferrin as a source of iron and investigated the mechanism by which iron can be obtained from this plasma protein. Adding either ferrous sulfate or holotransferrin to an iron-deficient culture medium allowed growth of S. intermedius. Cultivation of S. intermedius under an iron-poor condition was associated with the over expression of a 58 kDa cell surface protein. Neither siderophore activity nor reductase activity could be detected. Moreover, cells of S. intermedius did not show transferrin-binding activity or proteolytic activity toward transferrin. It was found that S. intermedius could rapidly decrease the pH of the medium during cell growth, resulting in a release of iron from holotransferrin. When the buffering capacity of the culture medium was significantly increased, the holotransferrin could not support growth of S. intermedius. It is suggested that under certain circumstances, S. intermedius may migrate from its normal niche (oral cavity), reach a particular site and create a localised environment where the pH can be lowered with the subsequent release of iron from transferrin. This would allow bacterial growth and initiation of the infectious process. PMID- 9741091 TI - Mitogen-activated protein kinase-defective Candida albicans is avirulent in a novel model of localized murine candidiasis. AB - Candida albicans strains with a deletion of the mitogen-activated protein kinase CEK1 gene are defective in the yeast to hyphal transition on solid surfaces in vitro. The virulence of a cek1 delta/cek1 delta null mutant strain was compared with its wild-type parent strain (WT) in a novel model of localized candidiasis. The mammary glands of lactating mice (at day 5 postpartum) were infected for 2, 4 and 6 days with 50 microliter suspension containing 1 x 10(5), 1 x 10(6) and 1 x 10(7) blastopores before death. Infected and non-infected control glands were evaluated pathologically. All animals infected with cek1 delta/cek1 delta null mutant strains showed no lesions while 65% of animals infected with the WT strain had severe lesions characterized by widespread heterophilic infiltration, necrosis, and abscess formation. As an additional control, animals infected with the disrupted strain complemented with the WT CEK1, on a replicating plasmid, also showed severe pathological changes similar to the WT strain. These results clearly demonstrate that the CEK1 gene codes for a virulence determinant of C. albicans and that the mouse mastitis model is well suited for the discriminative study of the pathogenicity of different C. albicans strains. PMID- 9741092 TI - TcDJ1, a putative mitochondrial DnaJ protein in Trypanosoma cruzi. AB - A full length cDNA encoding a novel Trypanosoma cruzi DnaJ protein was cloned and characterized. The 324 amino acid protein encoded by the cDNA (TcDJ1) displays a characteristics J-domain, but lacks the Gly-Phe and zinc finger regions present in some other DnaJ proteins. Relative to four other T. cruzi DnaJ proteins, TcDJ1 has an amino terminal extension containing basic and hydroxylated resides characteristic of mitochondrial import peptides. A T. cruzi transfectant expressing epitope-tagged TcDJ1 was generated and subcellular fractions were produced. Western blot analysis revealed that the protein has a molecular mass of 29 kDa and is found in the mitochondrial fraction. The expression of TcDJ1 is developmentally regulated since the levels of both mRNA and protein are much higher in epimastigotes (replicative form) than in metacyclic trypomastigotes (infective form). Thus it may participate in mitochondrial biosynthetic processes in this organism. PMID- 9741094 TI - Enterococcus faecalis glutathione reductase: purification, characterization and expression under normal and hyperbaric O2 conditions. AB - Glutathione reductase is found ubiquitously in eukaryotes and Gram-negative bacteria, and plays a significant role in bacterial defense against oxidative stress. Glutathione reductase from the Gram-positive bacterium Enterococcus faecalis was purified to homogeneity using anion exchange, hydrophobic interaction, and affinity chromatography. The homogeneous 49-kDa enzyme contained 1 mol bound FAD per subunit. The determined N-terminal amino acid sequence of the E. faecalis enzyme displays significant identity with glutathione reductases from other Gram-negative and Gram-positive bacteria, as well as yeast and human erythrocyte reductases. The kinetic mechanism is ping-pong, and the determined kinetic parameters exhibited by the E. faecalis glutathione reductase are similar to those found for glutathione reductases from yeast, Escherichia coli, and human erythrocyte. A two-fold increased expression of glutathione reductase activity and a three-fold induction of glutathione peroxidase activity were observed under hyperbaric O2 growth conditions without a corresponding change in the total glutathione and soluble thiol content. The difference in the expression of the enzyme, and its cognate substrate's intracellular concentration, under these conditions suggest that the gene encoding glutathione reductase is responsive to oxygen concentration, but that the genes encoding the glutathione synthesizing enzymes are not linked to an oxygen-sensitive promoter. PMID- 9741093 TI - Phylogenetic position and codon usage of two centrin genes from the rumen ciliate protozoan, Entodinium caudatum. AB - A lambda phage cDNA expression library was constructed from washed suspensions of the rumen ciliate protozoan, Entodinium caudatum, which had been maintained in an isolated, monofaunated sheep. The library was screened using an anti-E. caudatum antiserum raised in rabbits against sonically disrupted protozoa, DNA sequences for two centrins or caltractins, a subfamily of the EF-hand Ca(2+)-modulated proteins which are closely related, highly conserved cytoskeletal proteins, were identified and characterised. Their phylogenetic position was established relative to other centrin gene sequences. The two proteins showed homology to Paramecium tetraurelia centrins: 50 and 52% identities and 71 and 75% similarities in the protein sequence, over 99 and 110 amino acids fragments. Codon usage and indices revealed the E. caudatum follows universal codon usage, but with a restricted number of codons, and has a low G&C content. PMID- 9741095 TI - Biochemical separations by continuous-bed chromatography. AB - Innovations in column-packing media for biomolecule purification have progressed from large spherical, porous polysaccharide beads to advanced polymeric supports. Continuous-bed technology is a radical new technology for chromatography based on the polymerization of advanced monomers and ionomers directly in the chromatographic column. The polymer chains form aggregates which coalesce into a dense, homogeneous network of interconnected nodules consisting of microparticles with an average diameter of 3000 A. The voids or channels between the nodules are large enough to permit a high hydrodynamic flow. Due to the high cross-linking of the polymer matrix, the surface of each nodule is nonporous yet the polymeric microparticles provide a very large surface area for high binding capacity. This paper will demonstrate the properties and advantages of using a continuous bed support for high resolution biomolecule separations at high flow-rates without sacrificing capacity. PMID- 9741096 TI - Synthesis and purification of hydrophobic peptides for use in biomimetic ion channels. AB - The synthesis and subsequent purification of several hydrophobic peptides is described. These peptides include the 24-residue M3 transmembrane domain of the rat connexin 32 protein, a peptide sequence that contains only seven amino acids with hydrophilic side-chains (71% hydrophobic). Moreover, for comparison, a much smaller hydrophobic octapeptide, designed to exist with alpha-helical secondary structure, was also studied. Optimum conditions for the RP-HPLC purification of these peptides was dependent on peptide length and solubility properties. PMID- 9741097 TI - Reversed-phase high-performance liquid chromatography of virus-like particles. AB - A quantitative reversed-phase high-performance liquid chromatography (RP-HPLC) method has been developed to detect the L1 subunit protein from virus-like particles (VLPs) of human papillomavirus (HPV). The method utilizes heat treatment with a buffer consisting of 50 mM Tris, pH 8.0 containing 8 M guanidine HCl and 10% 2-mercaptoethanol to dissociate the VLPs into monomeric L1. Following dissociation, the sample is injected onto a C4 or C8 column. The L1 protein is eluted as a single, clearly resolved peak. Elution conditions have been optimized to enhance the separation of L1 from other contaminants. Based on spike recovery studies and sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis this method is suitable for quantitation of various partially purified in-process samples and can be used to facilitate purification process development. PMID- 9741098 TI - Detergent extraction of herpes simplex virus type 1 glycoprotein D by zwitterionic and non-ionic detergents and purification by ion-exchange high performance liquid chromatography. AB - Detergents (surfactants) are the key reagents in the extraction and purification of integral membrane proteins. Zwitterionic and non-ionic detergents were used for the extraction of recombinant glycoprotein D (gD-1) of herpes simplex virus type 1 (HSV-1) from insect cells infected with recombinant baculovirus. The highest yield was obtained with the two alkyl carboxybetaine detergents (N dodecyl-N,N-dimethylammonio)undecanoate [DDMAU, critical micelle concentration (CMC) = 0.13 mM] and (N-dodecyl-N,N-dimethylammonio)butyrate (DDMAB, CMC = 4.3 mM). Therefore these zwitterionic detergents were used as additives to the elution buffers in ion-exchange high-performance liquid chromatography (HPIEC) to purify gD-1 of HSV-1 from the extracts. The non-ionic detergent pentaethyleneglycol monodecyl ether (C10E5) that was used in earlier studies [R.A. Damhof, M. Feijlbrief, S. Welling-Wester, G.W. Welling, J. Chromatogr. A, 676 (1994) 43] was used for comparison. Two columns were used, Mono Q and Resource Q, at 1 and 5 ml/min flow-rates, respectively. The results show that the detergents DDMAU and C10E5 are superior to DDMAB, when the detergents were used as additives to the elution buffers at 0.2% (w/v). With 0.2% DDMAB in the eluent, purification of HSV gD-1 was not possible. Detergents with a high CMC may be less suitable as additives in elution buffers. HPIEC at flow-rates of 1 and at 5 ml/min showed satisfactory results. At 5 ml/min HSV gD-1 was mainly concentrated in two eluent fractions. The highest recovery of gD-1 was obtained either by chromatography of a C10E5 extract using a Mono Q column at a flow-rate of 1 ml/min or by chromatography of a DDMAU extract using a Resource Q column at a flow-rate of 5 ml/min. PMID- 9741099 TI - Real-time isoform analysis by two-dimensional chromatography of a monoclonal antibody during bioreactor fermentations. AB - A humanized monoclonal antibody specific for L-selectin exhibits two distinct isoforms that are distinguishable by a charged group modification on one of the antibody light chains. The added charge allows baseline separation of the isoforms by anion-exchange chromatography. Since this modification most likely results from specific enzymatic activity within the Golgi complex, it is possible that fermentation conditions may affect the relative amounts of the isoforms produced. Herein is described a two-dimensional chromatographic method for quantifying the relative amounts of the isoforms from fermentation broths, in real time. PMID- 9741100 TI - Characterization of the F(ab')2 fragment of a murine monoclonal antibody using capillary isoelectric focusing and electrospray ionization mass spectrometry. AB - We characterized a F(ab')2 fragment obtained by pepsin cleavage from a murine monoclonal IgG3 by means of electrospray ionization (ESI) mass spectrometry (MS), capillary isoelectric focusing (cIEF), high-performance anion-exchange chromatography-pulsed amperometric detection (HPAEC-PAD) and LC-MS peptide mapping. Separation of the fragment by cIEF under nonreducing conditions resulted in a number of distinct peaks. Using reducing conditions the heavy chain and the light chain were separated into two peaks each. Analysis by ESI-MS revealed a high mass heterogeneity of the molecule. Digestion with neuraminidase simplified both the cIEF pattern and the mass spectrum. The cIEF of the reduced molecule showed that the sialic acids were located only on the heavy chain of the F(ab')2 fragment. By incubation with O-glycosidase a further reduction of the complexity of the mass spectrum was achieved showing 8 different isoforms. By LC-MS peptide mapping these isoforms could be attributed to the heterogeneity of the pepsin cleavage site in the hinge region of the antibody. The sugars of the O-linked carbohydrate chain were identified by HPAEC-PAD as galactosyl-N-acetyl galactosamine (GalNAcGal) with terminal N-glycolylneuraminic acid. The glycosylation site was identified by peptide mapping and amino acid sequence analysis as Ser222. PMID- 9741101 TI - Hydrophilic interaction/cation-exchange chromatography for separation of amphipathic alpha-helical peptides. AB - Mixed-mode hydrophilic interaction/cation-exchange chromatography (HILIC/CEX) is a novel high-performance technique which has excellent potential for peptide separations. Separations by HILIC/CEX are carried out by subjecting peptides to linear increasing salt gradients in the presence of high levels of acetonitrile, which promotes hydrophilic interactions overlaid on ionic interactions with the cation-exchange matrix. In the present study, HILIC/CEX has been applied to the separation of synthetic amphipathic alpha-helical peptides, varying in amphipathicity and the nature of side-chain substitutions in the centre of the hydrophobic or hydrophilic face. Observation of the retention behaviour of these amphipathic alpha-helical peptide analogues during HILIC/CEX and reversed-phase chromatography (RPLC) enabled the establishment of general rules concerning the applicability of these complementary HPLC techniques to peptides displaying a secondary structural motif of common occurrence. PMID- 9741102 TI - High-performance ion-exchange chromatography and adsorption of plasma proteins. AB - Resolution and recovery are primary concerns in protein chromatography. Separations are often by size, ion exchange, or hydrophobic-hydrophilic properties of the support, eluent and protein. Adsorption of a protein to a synthetic surface plays an essential role in this complex process. In this study, we examined the adsorption properties of three representative plasma proteins (albumin, fibrinogen, and immunoglobulin G) on nonporous column materials containing either quaternary amine or sulfopropyl functional groups. The adsorption properties were studied at 37 degrees C and pH 7.4. Salt gradients were used to examine the adsorption/desorption properties of each of the proteins on each type of surface. The salt concentrations at desorption were measured and compared to the protein isoelectric points. In addition, we examined protein recoveries as a function of desorption time. Our results suggest that protein recoveries depend not only on the protein, eluent and surface, but also the residence time and overall charge concentration during the initial adsorption process. Finally, we correlated the number of charge sites on a molecule with the width of a chromatographic band at half height. The data produced as a result of this study may be used to determine the actual unfolding time of a protein, given a certain set of conditions. The data may also help in understanding the chemistry and dynamics of the protein adsorption processes in ion-exchange chromatography as well as provide key structural information about the proteins. PMID- 9741103 TI - Thin-layer ion-exchange chromatography of proteins. AB - Thin-layer chromatography (TLC) is one of the simplest and most convenient techniques to separate small molecules. Of a variety of TLC separation modes, only size-exclusion was successfully used to separate proteins. In this paper, adsorption-TLC was used to separate proteins. The net charges were calculated for four model proteins, albumin, transferrin, lactoferrin and lysozyme, under different pH values. The suitable pH values for separation were determined according to the results from such calculations. Then, the adsorption isotherms of the four proteins were measured to deduce the ionic strength for appropriate elution conditions. Optimal conditions, 0.01 M bicine and pH 8.50, and a three step elution process (1st step 0.01 M NaCl, 2nd 0.025 M NaCl, and 3rd 0.10 M NaCl), were obtained. Finally, the four model proteins were successfully separated under these elution condition. PMID- 9741104 TI - Recent developments of magnetic beads for use in nucleic acid purification. AB - The performance of Magarose, an agarose-based bead containing a paramagnetic component has been evaluated. The anion exchanger DEAE-Magarose is effective at binding DNA from a crude cell lysate. The plasmid pBluescript was isolated from 1.5 ml Escherichia coli JM109 cell culture, following alkaline lysis yielding 8.2 micrograms high-quality DNA. Under similar binding conditions 21 micrograms of salmon sperm DNA bound to the ion exchangers. The affinity medium oligo-dT Magarose was demonstrated to bind 75 mumol of an oligo-dA probe/g of medium by hybridization. Under similar conditions mRNA could be isolated from a preparation of baby hamster cell total RNA. The magnetic susceptibility of Magarose is very high, facilitating the use of this separation technique for rapid batch chromatographic processes. PMID- 9741105 TI - Properties of an alpha-galactosidase, and structure of its gene galA, within an alpha-and beta-galactoside utilization gene cluster of the hyperthermophilic bacterium Thermotoga maritima. AB - Thermotoga maritima represents one of the few hyperthermophilic bacteria currently known. The chromosomal alpha-galactosidase gene of T. maritima strain MSB8 has been cloned and its nucleotide sequence was determined. The gene, designated galA, has coding capacity for a 552 residue polypeptide with a calculated molecular mass of 63,653 Da. GalA was found to be flanked by other genes probably involved in galactoside breakdown and utilization. The previously sequenced beta-galactosidase gene, lacZ, is localized immediately upstream of galA while two open reading frames that putatively encode enzymes of galactose catabolism, i.e. galactose-1-phosphate uridylytransferase (galT) and galactokinase (galK), were found downstream of galA. The identified genes are extremely close together or even overlap and have the same orientation, so they could all be part of one galactoside utilization operon of T. maritima MSB8. GalA displayed low-level amino acid sequence similarity with alpha-galactoside of glycosyl hydrolase family 36. However, GalA is smaller than the other members of this enzyme family. The galA gene was expressed in Escherichia coli and the recombinant alpha-galactosidase was purified and characterized. The molecular mass of the recombinant enzyme was estimated at about 62 kDa by denaturting gel electrophoresis. Maximal hydrolysis of the chromogenic substrate p-nitrophenyl alpha-D-galactopyranoside was measured at pH 5.0-5.5 and 90-95 degrees C (5 min assay). Divalent cations were not required for activity. The enzyme released galactose from raffinose, melibiose and the synthetic substrates p-nitrophenyl and omicron-nitrophenyl-alpha-D-galactopyranoside. The T. maritima alpha galactosidase thus was highly specific for the galactose moiety and the alpha anomeric configuration of the glycosidic linkage. Its extreme thermal stability (t 1/2 = 6.5 h at 85 degrees C) makes this enzyme an interesting candidate for biotechnological applications. PMID- 9741106 TI - F-and V-ATPases in the genus Thermus and related species. AB - The discovery of a V-type ATPase in the gram-negative bacterium Thermus thermophilus HB8 (YOKOYAMA et al., J. Biol. Chem. 265, 21946, 1990) was unexpected, since only eukaryotic endomembranes and archaea were thought to contain this enzyme complex, and horizontal gene transfer was suggested to explain the finding. We examined membrane-associated ATPases from representatives of several groups of the genus Thermus. The enzymes were extracted with chloroform and purified by ion exchange chromatography or native gel electrophoresis. One novel Islandic isolate, T. scotoductus SE-1, as well as strain T. filiformis from New Zealand, possessed F-ATPases, as judged by the typical five subunit composition of the F1-moiety, sensitivity to azide, insensitivity to nitrate and a strong crossreaction with antibodies against the F1-ATPase from E. coli. In addition, N-terminal amino acid sequencing of the beta subunit from T. scotoductus SE-1 confirmed its homology with beta subunits from known F-ATPases. In contrast, the same extraction procedure released a V-ATPase from the membranes of T. thermophilus HB27 and T. aquaticus YT-1. The related species Meiothermus (formerly Thermus) chliarophilus ALT-8 also possessed a V ATPase. All V-ATPases examined in this study contained larger major subunits than F-ATPases, crossreacted with antiserum against subunit A of the V-ATPase from the archaeon Halobacterium saccharovorum, and the N-terminal sequences of their major subunits were homologous to those of other V-ATPases. Sequences of the 16S rRNA gene clearly placed T. scotoductus SE-1, along with other non-pigmented Thermus strains, as a distinct species close to T. aquaticus. Our results suggested that at least two members of the genus, T. scotoductus SE-1 and T. filiformis, contain an F-ATPase, whereas several others possess a V-ATPase. These data could indicate a greater diversity of the genus Thermus than was previously thought. Alternatively, the genus may consist of species where horizontal gene transfer has occurred and others, where it has not. PMID- 9741107 TI - First report on the identification of microcystin in a water bloom collected in Belgium. AB - A toxic cyanobacterial bloom dominated by Microcystis aeruginosa occurred in 1995 in three adjacent ponds near Liege (Belgium) where at the same time conspicuous bird deaths were observed. The toxicity assay using primary rat hepatocytes indicated a high hepatoxicity. A 4 h incubation yielded a LD50 of 0.23 mg bloom material (dry weight)/ml cell culture medium. Toxicity was due to hepatotoxins of the microcystin class, microcystin-LR and-RR being the major microcystins present as determined by RP-HPLC absorption spectra, 1H NMR, and ESMS spectra. Additionally, the bloom sample contained small amounts of microcystin-YR. The microcystin content of the dry bloom biomass was 870 micrograms/g (on the basis of the hepatotoxicity assay) and 556 micrograms/g (on the basis of the RP-HPLC peak area). A higher yield of microcystins was obtained by acetic acid extraction instead of methanol extraction, whereas different extraction temperatures (20 degrees C, 40 degrees C) had no effect on the yield. PMID- 9741108 TI - Bacillus subtilis develops competence for uptake of plasmid DNA when growing in milk products. AB - Transformation with plasmid DNA of naturally competent cells of Bacillus subtilis 168 in milk products was studied. Plasmid pMG36enpr, a broad host-range lactococcal vector carrying an erythromycin resistance and the B. subtilis npr gene encoding neutral protease, was taken up by B. subtilis cells grown in UHT chocolate milk. Under these conditions competence was optimal during transition from exponential to stationary growth phase, resulting in 9 x 10(1) transformants per 0.01 microgram DNA. No manipulation of the cells was necessary for competence to develop. When cells were pregrown in synthetic medium, higher transformation rates were obtained in assays, where the subsequent transformation experiments were either done in chocolate milk diluted 1:1 (v/v) with synthetic growth medium (up to 8 x 10(2) transformants) or in undiluted chocolate milk (1 x 10(2) transformants). The number of transformants was reduced to 4 x 10 (1), when diluted milk or flavored milks were used. No transformants were obtained in diluted yoghurt. Controls, in which both the preculturing and the transformation assays were done in synthetic medium, gave the maximum number of transformants (4 x 10(3) transformants per 0.01 microgram DNA). PMID- 9741109 TI - Isolation and characterization of Thermococcus barossii, sp. nov., a hyperthermophilic archaeon isolated from a hydrothermal vent flange formation. AB - A new hyperthermophilic microorganism, Thermococcus barossii, was isolated from rock fragments of a hydrothermal vent flange formation, located along the East Pacific Rise of the Juan de Fuca Ridge. This organism is obligately anaerobic and grows over a temperature range of at least 60-92 degrees C in artificial seawater based media, containing elemental sulfur, tryptone and yeast extract. The addition of a maltooligosaccharide mixture and tungsten to this medium improved growth to some extent. At the Topt for growth (82.5 degrees C), cell densities as high as 4 x 10(8) cells/ml could be obtained in 18-liter batch fermentations, with a doubling time of approximately 40 minutes, if culture access to elemental sulfur was sufficient. In continuous culture at the same temperature, comparable cell densities could be obtained but only at slower growth rates. Morphologically, T. barossii is coccoid-shaped, forming irregularly-shaped spheres; under optimal conditions, these coccoids become more regular and smaller, a characteristic of other hyperthermophilic archaea. Negatively-stained preparations showed no pili or flagella associated with the cell surface. 16S rRNA sequencing reveals that T. barossii is most similar to Thermococcus celer (99.7%). Yet, further comparisons with T. celer showed that T. barossii is a new Thermococcus species: different growth temperature optimum (82.5 degrees C vs. 88 degrees C), obligate requirement for sulfur, higher G + C content (60% vs. 56.7%) and 47.7% DNA-DNA hybridization. The nucleotide and translated amino acid sequence for the gene encoding a DNA polymerase from T. barossii was compared to sequences of related genes from other Thermacoccales. The polymerase phylogenies were congruent with those obtained from the 16S rRNA phylogenetic analyses. Based on the high degree of similarity among members of the genus Termococcus for the criteria used thus far, aspects of enzymology may be an important mechanism of differenting one species from another. PMID- 9741110 TI - Characterization of atypical Aeromonas salmonicida by different methods. AB - Fifty two isolates of atypical Aeromonas salmonicida, recovered from a wide range of hosts and geographical locations, were heterogeneous in terms of molecular and phenotypic characteristics, and represented taxa which could not be accommodated by the current classification of four subspecies. Generally, there was incongruence between the molecular (PCR, RAPD and ribotyping) and phenotypic methods in terms of cluster membership. By PCR, 6 groups were described of which Group 1 encompassed 12 isolates including the type strain of A. salmonicida subsp. smithia. Group 2 accommodated 23 isolates including the reference cultures of subspecies achromogenes and masoucida. The named culture of Haemophilus piscium was recovered in Group 6. By ribotyping and RAPD, the reference cultures were recovered in separate groups. All methods pointed to the uniqueness of subspecies smithia. Most isolates contained 2-6 plasmids, of 2.3 to 150 kb in length. Nevertheless, all isolates possessed certain key characteristics, including Gram-negativity, and the absence of motility. PMID- 9741111 TI - Design and evaluation of a 16S rRNA-targeted oligonucleotide probe for specific detection and quantitation of human faecal Bacteroides populations. AB - Colonic Bacteroides include several species which, by their population level and activities, are significant contributers to the metabolic activity and health of man and animals. Yet, the understanding of their ecology has been hampered by the lack of highly specific and reliable enumeration techniques. Based on 16S rRNA sequence comparisons within the available database, we have designed an 18-mer oligonucleotide that targets a region common to-and specific for the Bacteroides Porphyromonas-Prevotella group. We have tested the specificity of the probe and its usefulness for studies of human faecal samples. Under experimentally optimized hybridization conditions, the probe was shown to similarly recognize the rDNA obtained from 40 strains representing 8 species of the Bacteroides Porphyromonas-Prevotella group. Importantly, it did not recognize 31 strains of microorganisms representing 8 genera of the dominant human faecal microbiota. Among selected colonies of dominant microorganisms of the faecal flora of two human individuals, strains identified as B. vulgatus by immunoblots using a species-specific monoclonal antibody were all detected by the probe. Colony hybridization was used to enumerate total Bacteroides-group microorganisms in faecal specimen from children and adults. The probe described therein was further used in quantitative RNA blots to monitor fluctuations of the Bacteroides-group versus Bifidobacterium genus in frozen faecal samples from a child between 85 and 125 days of age. It will be applicable to similar investigations of other anaerobic environments. PMID- 9741112 TI - A qualitative evaluation of the published oligonucleotides specific for the 16S rRNA gene sequences of the ammonia-oxidizing bacteria. AB - Over the past few years, there has been an increasing interest in making oligonucleotides specific for ammonia-oxidizing bacteria (AOB), in order to detect and monitor these slow growing bacteria in environmental samples, in enrichment cultures and in wastewater treatment plants. Based on 16S rDNA sequences, a broad selection of oligonucleotides have been designed, either encompassing all known AOB in the beta-subgroup of the Proteobacteria (beta AOB), or subclasses within beta AOB. Thirty different oligonucleotides have so far been published, with varying specificity. The first AOB-specific oligonucleotides published were obtained as a result of an alignment of only eleven 16S rDNA sequences from AOB. Including the present study, there are now forty nearly full length 16S rDNA sequences available from these bacteria, in addition to a number of partial sequences, so that an improved evaluation of the published oligonucleotides can be done. Two new 16S rRNA gene sequences from Nitrosospira are presented here, in a phylogenetic analysis containing every 16S rRNA gene sequences (> 1 kb) available from AOB. On the basis of an alignment of all these sequences, combined with searches in the nucleotide sequence databases, an evaluation of the thirty published oligonucleotides is presented. The analysis expose the strength and weakness of each oligonucleotide and discuss the use of oligonucleotides specific for 16S rRNA genes in future studies of AOB. The present work also identifies one new, broad range primer, specific for the AOB in the beta-subgroup of the Proteobacteria. PMID- 9741113 TI - Identification of Leptospira inadai by continuous monitoring of fluorescence during rapid cycle PCR. AB - Seven new Leptospira isolates from rats, a buffalo, and contaminated media showed either reactive serology against more than 1 serogroup or no reactive serology against a reference panel of 22 serovars in the microscopic agglutination test (MAT). Because of these inconclusive results, the 16S rDNA sequences of these isolates were determined and found to resemble that of the type strain of Leptospira inadai (L. inadai), serovar lyme strain 10, which is considered to be nonpathogenic for humans. Comparative analyses of other Leptospira 16S rDNA sequences from databases revealed a L. inadai-specific signature sequence, against which an amplification primer was designed. This primer when used in conjunction with an universal primer enabled the trial of a rapid PCR protocol in which fluorescence emissions due to binding of SYBR Green I dye to PCR products were continuously monitored during rapid thermal cycling. A melting curve acquired immediately after PCR was used to distinguish the intended product. The thermal cycling and continuous monitoring of fluorescence emission were accomplished by the LightCycler; the whole procedure of 30 PCR cycles and melting curve acquisition required only 20 minutes. The primer achieved the required specificity, as the intended PCR product resulted only from 6 confirmed L. inadai reference strains and 7 field isolates that had been verified as L. inadai by the 16S rDNA sequencing, but not from 16 reference strains of Leptospira belonging to 7 other genospecies. Furthermore, these experiments showed that the PCR protocol was robust because target DNA of different conditions, which were extracted by either 1 of the 4 methods used, could be detected. PMID- 9741114 TI - Environmental distribution and diversity of Bacillus thuringiensis in Spain. AB - Bacillus thuringiensis was isolated from 301 out of 1,005 samples collected in Spain from agricultural and non-cultivated soils, dust from stored products, and dead insects. Based on the production of parasporal crystals, 1,401 isolates were identified as B. thuringiensis after examining 11,982 B. thuringiensis-like colonies. We found a greater presence of B. thuringiensis in dust from grain storages than in other habitats. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the spore-crystal mixtures revealed diverse populations of B. thuringiensis which were differentiated in at least 92 distinct protein profiles. Serological identification also showed great diversity among the Spanish isolates which were distributed among 38 of the 58 known serovars. The most frequently found serovars were aizawai, kurstaki, konkukian, morrisoni, and thuringiensis, which together represented more than 50% of the serotyped isolates. In preliminary toxicity assays, a number of isolates were found to show significant insecticidal activity against the lepidopterans Heliothis armigera (76.1% of the assayed isolates), Spodoptera exigua (50.5%), and Plutella xylostella (19.7%). Thirty five isolates were toxic to both H. armigera and S. exigua, and eight were toxic to S. exigua and P. xylostella. Four and one isolates were toxic to the coleopterans Leptinotarsa decemlineata and Colaspidema atrum, respectively, and three to the dipteran Tipula oleracea. The electrophoretic pattern and serovar of most of the isolates with toxic activity were consistent with those reported in the literature, although other isolates revealed unusual protein profiles, were assigned to new H serovars, or were included in H serovars not previously reported within such pathotypes. PMID- 9741115 TI - Antibiotic resistance genes in coagulase-negative staphylococci isolated from food. AB - Coagulase-negative staphylococci were isolated from different raw milk cheeses and raw meat products and screened for their antibiotic resistances. They were identified as Staphylococcus xylosus, S. lentus, S. caprae, S. epidemidis and S. haemolyticus. The most frequent resistances found were those to chloramphenicol, tetracycline, erythromycin and lincomycin. They have been characterized on the molecular level. The chloramphenicol resistance genes were localized in several S. xylosus and S. caprae on plasmids with sizes ranging from 3.8-kb to 4.3-kb and were identified as chloramphenicol acetyltransferase (cat). All the tetracycline resistant strains were identified as S. xylosus and harboured a 4.4-kb plasmid carrying the tetracycline efflux resistance gene (tetK). The two erythromycin/lincomycin resistant S. caprae and S. epidermidis strains did not hybridize with the MLSB resistance genes ermAM, ermA, ermB and ermC. Three erythromycin resistant Staphylococcus sp. strains harboured an erythromycin efflux resistance gene (msr) localized twice on a 18-kb plasmid and once on the chromosome. A S. haemolyticus strain showing resistance to both lincomycin and clindamycin harboured a linA gene-carrying 2.2-kb plasmid. Further resistances to gentamicin, penicillin and kanamycin were less frequently observed and yet not characterized on a molecular level. PMID- 9741117 TI - A comparison of strategies for the detection and recovery of Vibrio vulnificus from marine samples of the western Mediterranean coast. AB - We have compared the effectiveness of culture-based methods and a DNA-based method for the detection, of Vibrio vulnificus from a seawater and three types of shellfish collected from the costal waters of Valencia, Spain. For culture-based method, we used two selective media, thiosulphate-citrate-salts-sucrose (TCBS), and cellobiose-polymyxin B-colistin (CPC) agars with and without previous enrichment in alkaline-saline-peptone-water (APWS). Presumptive colonies were confirmed as V. vulnificus by the polymerase chain reaction (PCR) using previously described 23S rRNA V. vulficus-specific sequences as primers (Dvu 9V and Dvu 45R). Direct detection was accomplished by a nested-PCR procedure developed for environmental samples, with the above mentioned primers for the second amplification. Of 32 seawater samples, only one yielded positive results by direct detection by PCR, whereas five were positive by culture methods. Of the 32 bivalve samples, two were positive by PCR and five by culture methods. From a total of 675 presumptive colonies selected on the two media, only 48 (20 from seawater and 28 from bivalves) were confirmed as V. vulnificus by PCR. Forty-six V. vulnificus isolates were obtained after enrichment and only two after direct inoculation of CPC. Except for one sampling, positive results by direct detection did not correlate with confirmed strains obtained from culture media. API 20E profiles were recorded for all isolates previously identified as V. vulnificus, revealing that around 20% of the strains were sucrose-positive. For our samples, the best strategy consisted in the combination of culture based methods (3 h enrichment in APWs at 40 degrees C followed by CPC at the same temperature) and DNA-based procedures (specific PCR amplification of the presumptive colonies with primers Dvu 9V and Dvu 45 R), which allowed the detection and accurate identification of V. vulnificus in less than 48h. This is the first report on the detection of cells of V. vulnificus naturally present in seawater and edible shellfish in the Spanish Mediterranean coast. PMID- 9741118 TI - Isolation and characterisation of obligately anaerobic, lipolytic bacteria from the rumen of red deer. AB - Two Gram-positive, obligately anaerobic, lipolytic bacteria, isolates LIP4 and LIP5, were obtained from the rumen contents of juvenile red deer. These mesophilic bacterial strains were capable of hydrolysing the neutral lipids, tallow, tripalmitin and oliver oil, into their constituent free long-chain fatty acid and glycerol moieties. The latter compound was dissimilated by both isolates, with isolate LIP4 producing propionate as the predominant product, while isolate LIP5 produced acetate, ethanol and succinate. The lactate-utilising isolate LIP4 grew on a limited range of saccharide substrates including glucose, fructose and ribose, and exhibited an unusual cell wall structure and morphology. The isolate LIP5 grew upon a wider range of saccharides, but was unable to use lactate as a substrate. Based upon phenotypic and 16S rRNA gene sequence analyses, isolate LIP4 clusters with species in the genus Propionibacterium, while isolate LIP5 is a member of clostridial cluster XIVa. PMID- 9741119 TI - A novel isolate of Bacillus thuringiensis serovar leesis that specifically exhibits larvicidal activity against the moth-fly, Telmatoscopus albipunctatus. AB - A soil isolate designated 88-KO-14-45, belonging to Bacillus thuringiensis serovar leesis (H33), exhibited larvicidal activity against the moth-fly, Telmatoscopus albipunctatus (Diptera: Psychodidae), but not for larvae of the culicine and aedine mosquitoes and Lepidoptera. Purified parasporal inclusions had an LC50 value of 5.78 micrograms/ml for the larval moth-fly, but gave no mortality against larvae of Culex pipiens molestus (Diptera: Culicidae) at protein concentrations up to 10 mg/ml. Electron microscopic observations revealed that the parasporal inclusions are homogeneous round-shaped bodies enclosed with thick, electron dense envelopes. Haemolytic activity against sheep erythrocytes was not detected in the solubilized inclusions. SDS-PAGE showed that the inclusions are composed of 72, 68, 56 and 30 kDa proteins. Immunologically, these proteins were unrelated to the inclusion proteins of B. thuringiensis serovar israelensis, while a 70 kDa protein of the strain 73-E-10-2 (B. thuringiensis serovar darmstadiensis) was seroactive to antibodies against proteins of 88-KO-14 45. PMID- 9741120 TI - Characterization of bacteriocins produced by strains from traditional Bulgarian dairy products. AB - A result of extensive screening of over 300 strains from the Collection of ELBY Bulgaricum, PLC, thirty six strains were selected as producers of bacteriocins, active closely related lactic acid bacterial species and some food spoilage bacteria. The selected strains belong to L. helveticus, L. bulgaricus and S. thermophilus, which are rare bacteriocin producers. Nineteen nonidentified producers were characterized by molecular taxonomic approaches--M13 fingerprinting, repetitive PCR, ribotyping and hybridization with species specific probes, which allowed to affiliate them to the species L. delbrueckii. Several strains were found to harbour plasmids of different size. The estimated activity against food borne pathogens makes the isolated substances perspective as safe food preservatives and the producing strains could be used as components of starters with improved quality. PMID- 9741121 TI - Serotype H5a5b is a major clone within mosquito-pathogenic strains of Bacillus sphaericus. AB - Seventy six mosquito pathogenic strains of Bacillus sphaericus and 10 non pathogens were examined by pulsed field gel electrophoresis (PFGE) of SmaI digested chromosomal DNA. Non-pathogenic strains were clearly distinguished from the entomopathogenic types which were assigned to 21 groups (SmaI restriction patterns; SRPs). Some agreement between SRP based on PFGE and serotyping was noted, in particular all 39 strains of serotype 5a5b examined revealed identical SRPs indicating total conservation of the SmaI restriction site in these bacteria. Serotype 5a5b (SRP 12) strains comprise a widely distributed and abundant clonal lineage. Most serotypes, however, were divided into several SRPs. Seven strains from serotype 2a2b were covered in five SRPs in which toxin synthesis was correlated with chromosomal structure. Similarly, toxicity correlated with SRP in strains from serotypes 3 and 6. PMID- 9741122 TI - [Current concepts in maxillo-facial surgery]. AB - The authors make a brief and concise overview of the evolution of maxillofacial surgery over the last decades namely the concepts and methods used. Some case reports are presented which illustrate the application of the current methodology with the purpose of covering various fields of application (traumatologic, othognatic and tumoural). The need for an interdisciplinary assessment of these patients is discussed. PMID- 9741123 TI - [Facial paralysis. Neuro-muscular reconstruction techniques]. AB - Several surgical procedures have been proposed through the years for the treatment of facial paralysis. The multiplicity and diversity of techniques portray the complexity and challenge represented by this pathology. Two basic dynamic options are available: -Reconstruction of nerve continuity through direct micro suture, with interposition grafts or nerve transpositions. -Regional muscular transposition, most often using the temporalis. Facial reanimation with the temporalis transfer has withstood the test of time and still is a reference technique. In a few weeks, good results can be obtained with a single and rather simple surgical procedure. Functional free flaps have been used with increasing frequency in the last two decades, most often combining a cross-facial nerve graft followed by a gracilis free flap nine months later. With this method there is a potential for restoration of spontaneous facial mimetic function. Apparently there is a limit in microsurgical technique and expertise beyond which there is no clear improvement in nerve regeneration. Current research is now actively studying and identifying nerve growth factors and pharmacological agents that might have an important and complementary role in the near future. PMID- 9741124 TI - [Plastic and reconstructive surgery in oncology]. AB - The authors are members of the surgical staff of a central hospital specialized in the treatment of cancer (Portuguese Institute of Oncology-Francisco Gentil, Lisboa). They present their experience as plastic and reconstructive surgeons, mostly in the use of current reconstructive possibilities, after excision surgery or in a later phase. The aim is to offer the best quality available, which is evaluated by morbidity, mortality and quality of life. Most of the patients are subjected to different series of oncologic therapy, which is submitted to a multidisciplinary decision. PMID- 9741125 TI - [Microsurgical reconstruction of the upper extremity. Experience with 24 free flaps]. AB - The authors present the experience of 24 clinical cases of microsurgical reconstruction of the upper limb. These reconstructions were accomplished by different microvascular free flaps which were selected according to the type, size and location of the defects, having applied fasciocutaneous, adipofascial, myo, myocutaneous, tendofasciocutaneous, omentum flaps and 2nd toe transfers. The etiology was traumatic in 23 cases and tumoral in 1 case. A careful analysis was made for surgical times, selection and survival rate of the free flaps and the morphofunctional quality of the reconstructions. Microvascular free flaps are considered the choice technique for upper limb reconstruction, particularly for the hand in certain defects, considering the better anatomic and functional quality of the reconstructions. PMID- 9741126 TI - [Revascularization and reimplantation of the upper extremity. 15-year-experience with 114 cases]. AB - Progress in micro or macro replantation has resulted in higher survival rates of formerly amputated parts. More amputated digits or limbs survive because the time of ischemia can be exceeded by using cold storage or perfusion. Homo or heterodigital vessel transposition, expanded indications for vein graft interposition, as well as heterotopic transplantation allow for extremity preservation even in crush injuries, and in disastrous multiple amputations combined with contusion or avulsion. Secondary reconstruction with regard to bone defects, tendon repair, and eventual nerve grafting have to be aspired, finally leading to an improvement of functional results in daily and leisure activities as well as in early professional readaptation. A total of 114 microvascular extremity replantations/revascularizations with a survival rate of 77.2% were followed for an average of 15 years. PMID- 9741127 TI - [Rheumatoid pannus of the wrist]. AB - Presentation of the destructive mechanisms provoked by rheumatoid pannus at the level of ligaments, joints and bones of the wrist. Evaluation of the wrist problems requiring surgical solution, in particular at the radio-carpal and distal radio-ulnar joints. Review of the most useful surgical techniques for soft tissue and bone surgery. PMID- 9741128 TI - [Microsurgery in breast reconstruction. Myocutaneous free flap from the anterior rectus abdominis]. AB - The advantages of the free TRAM flap over the conventional Tram flap are known. The use of its main pedicle--the deep inferior epigastric system--improves the blood supply, decreasing the risk of skin and fat necrosis. The harvesting of 5-7 cm of muscle, and the preservation of its lateral border decreases the risk of abdominal wall bulge or hernias. Delayed breast reconstructions in patients submitted to radiotherapy were performed by end to side anastomosis between flap vessels and axillary vessels, avoiding the thoracodorsal irradiated vessels, and improving the blood flow. Ten patients were submitted to breast reconstruction by free TRAM flaps. There was one total flap necrosis, and one delayed healing around the periumbilical suture. Neither skin nor fat necrosis were seen. One patient developed an abdominal wall bulge. Two patients presenting tumor metastasis abandoned the plastic surgery outpatient clinic. Two patients refused the nipple-areolar complex (NAC) reconstruction. The outcome of five NAC reconstructions was very good, breasts being symmetrical without an opposite breast operation. PMID- 9741129 TI - [Agenesis of the vagina]. AB - In the last 10 years we treated ten patients with the diagnosis of vaginal agenesia. The reconstruction was performed according to the McIndoe procedure. The medical records of these patients were retrospectively reviewed in what concerns the diagnosis (Mayer-Rokitansky S.-7, Androgenital S.-1, Testicular feminization S.-1), classification, treatment, complications, and outcome. It is the authors' opinion that the McIndoe method is the most appropriate for the treatment of vaginal agenesia, because of its simplicity, low morbidity and absence of mortality. PMID- 9741130 TI - [Epidemiologic study of 1768 burned patients]. AB - A retrospective epidemiological study was made based on 1,768 burn patients admitted either to the Burns Unit or the general wards of the Plastic Surgery Department of Prelada Hospital in Oporto, from October 1988 until November 1997, corresponding to almost 8.5% of the overall patients admitted. The average age of the patients was 31.7 years and nearly 42% were children under 15 years old. The majority were male (60.8%), both in adults and children. The most frequent etiological agent was caused by fire (44.2%) and scalds (39%). About 74% of the patients required surgical treatment, with an average of nearly 2.3 operations per patient, of which 41.6% were surgical debridement or escarectomies and 40.8% skin grafting. The average hospital stay was 27.2 days and the overall mortality rate was nearly 6.8%, being much higher in adults (10.8%). PMID- 9741131 TI - [Contributions to the history of burn treatment]. AB - The authors make a brief review of burn treatment in Portugal as well as describing the Burn Unit of the Coimbra University Hospitals. The scientific advances in this area and new perspectives to the future are also mentioned, emphasizing the importance of burn prevention and a complete social reintegration of the burn patient. PMID- 9741132 TI - [Esthetic surgery of the facial bones]. AB - Aesthetic surgery of the facial skeleton deals with the developmental facial deformities, but does not include the treatment of congenital malformations or trauma. This paper reviews several aspects of nose and jaw aesthetic surgery and is illustrated with some clinical cases. PMID- 9741133 TI - [Hair transplantation. Past, present, and future]. AB - This paper briefly reflects the author's ideas regarding hair transplantation based on 25 years of experience in this field. Hair transplantation is a technique that moves hair from the donor dominant scalp to the recipient scalp with a variety of methods and protocols. The first written report on this surgical art form was made by Sasgawa in 1930, who described an insertion technique. Okuda later wrote about punch autografts and homografts in humans and animals in 1939; after which time Tamura and Fujita demonstrated reconstructive techniques with small grafts. However, it was not until 1959 that modern-day hair transplantation began, when Orentreich published a paper describing the theory of donor dominance. Recently, the advent of mini- and micrografts has indeed refined the technique, especially in those with early alopecia, in women and in facial scar alopecias. PMID- 9741134 TI - [Torsade de pointes: a new way to kill drugs]. PMID- 9741135 TI - [How to take care of your health. Education and promotion]. PMID- 9741136 TI - [Insulin resistance (IR) and obesity: role of leptin]. PMID- 9741137 TI - [The Madrid Declaration. A new ethics code for psychiatry in the 21th Century. Psychiatry World Association]. PMID- 9741138 TI - [Reproduction: role of the gametic structures in the success of fertilization. Simplification of the process]. PMID- 9741139 TI - [The role of positron tomography in suspected tumor recurrence based on the increase of serum markers]. PMID- 9741140 TI - [Natural medicine--balneologic therapies]. PMID- 9741141 TI - [Identification of the conductor of the neuronal orchestra who is responsible of the paradoxical sleep]. PMID- 9741142 TI - Morphological investigations of the surface epithelium of ductuli efferentes of black isogenic mice (Mus musculus). AB - Morphological investigations of the epithelial cell types that line the ductuli efferentes (DE) of black isogenic mice confirm absorption of the luminal fluid phase by endocytosis as the main function of ductuli efferentes (DE) in this species. Furthermore, all the histochemical and ultrastructural observations on the DE epithelial histoarchitecture indicate other cellular functions such as exocytosis and probably secretion, including an aprocrine secretory process. PMID- 9741143 TI - [The development of the horse testis]. AB - The aim of the study was to answer the open questions concerning the development of the horse's testis. This study revealed that the seminiferous tubules originate from the sex cords of the coelomic epithelium and Leydig cells from the proximal part of mesonephric nephrons, whereas the rete and the ductuli efferentes derive from intermediate and distal parts of the mesonephric tubules. During the development the Leydig cells undergo an enormous proliferation due to the PMSG secretion in the mare. The proliferation of these cells prevent the deep penetration of the rete into the medulla and is therefore the reason for the reduced extension of the rete and mediastinum testis in the stallion, although 80% of these cells degenerate in the last third of pregnancy. The growth of the seminiferous tubules during sexual maturity reduces the rete to the extremitas capitata of the testis. PMID- 9741144 TI - Muscle development in gilthead sea bream (Sparus aurata, L.) and sea bass (Dicentrarchus labrax, L.): further histochemical and ultrastructural aspects. AB - The histochemical profiles--mATPase and NADH-TR reactions--of the red and white muscle fibres of gilthead sea bream and sea bass were determined from the first week after hatching. Modifications of the mATPase technique by combinations of pH/time/molarity were carried out in order to compare the sensitivity of the myosin ATPase of each muscle fibre type of the lateral muscle. Results showed that the staining of muscle fibres was independent of small modifications in the technique. The intermediate 'pink' muscle was histochemically defined towards the end of the larval life and is considered to be implicated in the growth of the myotome. A layer of external cells was observed, by electron microscopical examination, between the connective tissue of the skin and the superficial red muscle fibres of larvae and postlarvae. It is suggested that the external cells are unlikely to be a source of red muscle fibres and implicated on the growth of the myotome, but rather a part of the dermatome. The timing, areas and mechanisms of hyperplastic growth of the myotome were defined and discussed. PMID- 9741145 TI - [Morphometric study of the wall of the caudal vena cava after the implantation of a filter in the sheep]. AB - The long stabilizers of the VT-LGM filter rest on large areas of the vascular wall. The morphometric study of the layers of the vascular wall, after implantation of filter in 15 ewes, divided into 3 groups of follow-up (2, 4 or 8 weeks) of 5 animals, was made at 3 levels opposite the filter and 1 level outside of it. Changes are seen for all the layers. Filter produces intimal hyperplasia. Stabilizers are quickly isolated from the blood flow. The thickness of the intimal hyperplasia grows for 4 weeks. It is more important at the base of the filter than at its head. There is also hyperplasia of the media with no change according to the follow-up or the level opposite the filter. The adventitia becomes thinner without variation of time or level of the filter. Smooth muscle cells of the adventitia become less frequent and smaller. Their density in front of the stabilizers, is the smallest by 4 week follow-up and remains the same along the stabilizer. The full thickness of the wall is bigger opposite the stabilizers than between them. The filter produces changes that are limited in time and in space. The most important changes are seen at 4 weeks after insertion and opposite the stabilizers. PMID- 9741146 TI - The morphology of the ureter in the duck (Anas platyrhynchos). AB - The morphology of the ureter of the duck was investigated, using histological, SEM and TEM techniques. The inner perimeter, the total thickness of the ureteral wall and the thickness of each uretral layer were measured. The epithelium was tall columnar and pseudostratified along all the tracts of the ureter and showed a high muco-secretive activity. The lamina propria contained numerous capillaries and aggregates of leucocytes and macrophages. Throughout the lamina propria there was a dense plexus of nerves. Some denuded single nerve fibres were observed between the epithelial basal cells. A plexus of nerves was also observed in the tunica muscularis. The ostium cloacale ureteris opened on a well-developed papilla in the dorsal region of the urodeum. The total thickness of the ureteral wall, the thickness of the lamina propria and the tunica muscularis, and the inner perimeter progressively increased towards the ostium. The above observations suggest that the avian ureter plays an important role in the modification of the urine coming from medullary cones, and in emission of the urine into the cloaca. PMID- 9741147 TI - Anatomicohistological characteristics of female genital tubular organs of the South American nutria (Myocastor coypus). AB - Morphohistological features of the tubular organs of the Myocastor coypus (coypu) female reproductive tract were studied. Specimens came from breeding farms with yard breeding systems. The analysis of the organs was made by histological methods and by macro and microscopic measurement techniques. The animals showed oviducts with macro and microscopically differentiable regions. Their inucosa showed primary branched folds in the ampullar sector. In the rest of the oviduct tract these folds were only of the primary type. The double uterus showed regional variations in the lumen, endometrial glands along the whole surface and a wide fibromuscular cervix with pseudoglands. The endocervical mucosa made clear a complex system of folds covered by a mucus-secreting epithelium. In the long vagina of the coypu a folded, rugose and irregular mucosa was observed. PMID- 9741148 TI - Detection of the Ki-67 proliferation associated nuclear epitope in normal canine tissues using the monoclonal antibody MIB-1. AB - This report describes the immunohistochemical detection of the Ki-67 proliferation associated nuclear epitope by means of the MIB-1 monoclonal antibody (mAb) in paraffin tissue sections from nine samples of 16 tissues obtained from nine dogs. Three parameters were considered: the localization of the cells expressing the Ki-67 epitope, the cytological characteristics of the Ki 67 expression, and the proliferative activity of some of the tissues measured by means of the proliferative index (percentage of Ki-67 positive cells measured on 500 and 1000 cells). The MIB-1 mAb reacts with the nuclei of proliferating cells, as in humans. The proliferative index was far from representative, but we were able to give a range of values concerning the proliferative activity of normal tissues. This study serves as a basis for the expression of the Ki-67 antigen by normal canine tissues in order to visualize the proliferative compartments and, thus, allows its application as a proliferative marker in routine veterinarian histopathology. PMID- 9741149 TI - Morphologic peculiarities of the renal cortical vasculature connected with blood redistribution in the kidney of the domestic pig. AB - The cortical blood vessels from 2 to 8 month old Camborow hybrid pigs and Belgium landrace pigs were studied using histologic and semithin sections as well as corrosion casts and cleared preparations of intravascular injection. It was established that the cortical blood vessels of the porcine kidney possess peculiarities that may regulate the redistribution of blood in the cortex. These peculiarities were found in the interlobular arteries, in the efferent arterioles of midcortical nephrons, in afferent arterioles of juxtamedullar glomeruli, and in the glomerular capillary networks. It was established that the number of renal corpuscles is different in the various zones of the cortex. PMID- 9741150 TI - Magnetic resonance imaging of the normal canine larynx. AB - The purpose of this study was to define the normal anatomic structures in the canine larynx with magnetic resonance images. TI-weighted images were taken in the sagittal and transverse planes. The MR images were obtained comparing MR images to dissection planes. Magnetic resonance imaging provides excellent anatomic detail of laryngeal structures. Therefore, it is of value of diagnostic imaging of some respiratory diseases in the dog. PMID- 9741151 TI - [The morphology of the thyroid gland in poultry with special regard to seasonal variations]. AB - The thyroid glands of 31 chickens at the age of 17 to 24 months were investigated. Different methods of anatomical preparation, casts of vessels and scanning electron microscopy were used. The thyroid gland of birds is a paired organ. It is located on the ventral surface of the base of the neck within the thoracic inlet. The left thyroid gland is placed more cranially than the right one. Each thyroid gland is closely connected to the common carotid artery on the medial side, from which it is supplied and to the jugular vein on the lateral side. It is a reddish-brown organ and of lenticular profile. The gland measures on average 10 mm in length, 6 mm in width and 2 mm in thickness, and is covered by a thin connective tissue capsule which holds adipose cells. It seems that each thyroid follicle is surrounded by a net of capillaries. The investigation by scanning electron microscopy proved that the follicles are oval with a pyramidal top on each end. The cuboidal epithelium cells leave impressions in the colloid. Epithelium cells carry microvilli on the follicle side surface. Described seasonal changes of the thyroid gland in size and activity were able to be confirmed by the examination of the organ in July and December. In winter the follicular cells were higher and the follicles had a greater volume. PMID- 9741152 TI - Histomorphometric and progesterone receptor immunohistochemical analysis in the oviduct of newly hatched chicks treated with follicle-stimulating hormone during embryonic development. AB - In this study we evaluated the histomorphology and ultrastructure of the oviduct of newly hatched chicks, as well as the immunohistochemical expression of progesterone receptor (PR) in this tissue after follicle-stimulating hormone (FSH) treatment on days 13, 15 and 17 of embryonic development. Results indicated a marked difference in the histology of the oviduct of newly hatched chicks treated with FSH. Magnum mucosa from these animals presented a pseudostratified epithelium with evaginations from the lumen into the epithelium and from the latter into the stroma beneath where tubular glands are formed. In contrast magnum mucosa from control animals presented columnar epithelium with no evaginations. In magnum epithelium FSH also induced the formation of cilia and microvilli projections into the lumen as well as an increase in the wall and lumen areas and in the density of nuclei per unitarea. PR immunoreactivity was only observed in the oviduct of FSH treated animals. PR was located in the nucleus of epithelial luminal cells, mucosal stromal cells and smooth muscle cells. These findings suggest that FSH induces an adequate hormonal milieu for the cytodifferentiation and PR gene expression in the chick oviduct. PMID- 9741153 TI - [The lists of non-subsidized medicines: an inevitable restriction?]. PMID- 9741154 TI - [HIV: preventive load and primary care intervention]. PMID- 9741155 TI - [Peripheral arteriopathy in type 2 diabetes mellitus]. AB - OBJECTIVES: To find the prevalence of peripheral arteriopathy (PA) in type 2 diabetics registered at a Health Centre, to detect their main risk factors and examine the usefulness of the portable Doppler in Primary Care consultations. DESIGN: A crossover descriptive study. SETTING: Urban Health Centre. PATIENTS: All the type 2 diabetics registered. MEASUREMENTS AND RESULTS: Anamnesis, physical examination, analysis and base ECG were performed. A portable Doppler determined systolic blood pressure in the lower extremities (foot and posterior tibial arteries) and brachial arteries in order to calculate the ankle/arm index (AAI). PA well recorded in the clinical history and AAI < or = 0.90 were considered criteria of PA. Diabetics with AAI > or = 1.25 were analysed separately. 289 patients with an average age of 65.3 (+/- 10.8 SD) were studied. 45.7% were men and 67% had Diabetes Mellitus for less than 10 years. 37.4% followed a dietary treatment and 21.1% were treated with insulin. Multivariate analysis showed that risk factors were: tobacco dependency, age, Hypertension and the type of treatment for the DM. CONCLUSIONS: The prevalence of PA found (21.4%) was very much higher than what had been previously diagnosed (6.9%). Identified risk factors were tobacco dependency, age, Hypertension and the type of DM treatment. A portable Doppler is easy to handle and allows peripheral arteriopathy to be diagnosed at an early stage. PMID- 9741156 TI - [Vaccination for the hepatitis B virus in primary care health staff: prevalence, affecting factors and need]. AB - OBJECTIVE: To find out the extent of HB vaccination in Primary Care health workers. DESIGN: An observational crossover study. SETTING: The Santiago de Compostela Health Area. PARTICIPANTS: All the Primary Care health staff in the area (N = 475). INTERVENTION: Data collection by means of a questionnaire sent by post to the entire target population of the study. MEASUREMENTS: Data analysis by means of logistic regression, and comparison of means and proportions. RESULTS: 65.6% took part. 74.0% of these stated that they had been exposed to risk at work during the month prior to the survey. 47.1% had received at least three doses of vaccine, 5.7% started their vaccination without completing it and 47.1% had never been vaccinated. Of this last group 4.8% suffered or had suffered the disease, leaving 42.3% who could be vaccinated. CONCLUSIONS: The present system of vaccination, in which the worker has to take the initiative, is not very efficient, even in highly motivated groups who are well aware of the risk. Vaccination programmes should be designed which offer the vaccine to all those healthworkers who can be vaccinated in their place of work. PMID- 9741157 TI - [HIV infection in the alternative centers for voluntary detection of anti-HIV antibodies in Cataluna, Spain (1995-1996). VIHDEVO collaborative group]. AB - OBJECTIVE: The knowledge of HIV serostatus may help the treatment and follow up of those infected people, and change the risky behaviours in those not infected. Epidemiological information from people tested can better address the activities of control and prevention of HIV infection. DESIGN: Collection of demographic and epidemiological information. PARTICIPANTS: People voluntary tested in four alternative test settings in Catalonia. MEASUREMENTS AND MAIN RESULTS: Of 1,733 petitions of voluntary testing, 63 (3.7%) were HIV positive. Overall prevalence in men were two fold than in women (4.6% vs 2.3%). In both years of study, the mean age for women HIV positive were higher than the mean age for women with aids. CONCLUSIONS: The results of this study confirm the age and sex pattern found for the HIV infection in other sentinel populations in Catalonia. Some measures should be taken in order to increase the accessibility of young women to the test. PMID- 9741158 TI - [Differences between knowledge and attitudes to AIDS in adolescents]. AB - OBJECTIVES: To relate knowledge about AIDS to the worry and fear caused by normal everyday situations, in which someone seropositive or with AIDS is involved, but when the virus cannot be transmitted. To find the discrimination that exists towards HIV carriers and people with AIDS. DESIGN: A basically descriptive crossover observation study. SETTING: Educational centres in the city of Castellon. PARTICIPANTS: 628 adolescents found by multi-stage conglomerate sampling. MEASUREMENTS AND RESULTS: A self-filled questionnaire was used. The items involving behaviour through which the virus was not transmitted, and for which the knowledge, worry and fear generated were evaluated, were described analytically. Items concerning discriminatory attitudes were analysed. Although we detected a lack of knowledge, the worry and fear generated by each situation was greater. In over 75% of those polled we observed discriminatory attitudes. CONCLUSIONS: We detected important divergences between knowledge about paths of non-transmission, and the anxiety and fear generated. Perhaps the preventive campaigns have contributed knowledge about the paths of contagion, but it is clear that they have not helped to modify discriminatory attitudes towards people with HIV or AIDS. PMID- 9741159 TI - [Level of knowledge about nourishment/nutrition among school adolescents in Cadiz]. AB - OBJECTIVE: To determine the level of knowledge about nourishment/nutrition and how this knowledge is distributed among school adolescent population in the town of Cadiz. DESIGN: Descriptive and transversal study. SETTING: Schools. PARTICIPANTS: Sample of 630 subjects from the school adolescent population in the town of Cadiz. MEASUREMENTS AND MAIN RESULTS: The average level of knowledge about nourishment/nutrition is 6.63 (in a 0-13 scale). No significant differences were found according to the perception of proportion or disproportion in height and weight, level of concern about body fats and getting fat, diets, avoiding some food or taking some medication, dietary fibers and infusions or any other weight-reducing products and physical exercise and fitting. The relationship between the level of knowledge and the BMI is very close to statistical significance. CONCLUSIONS: We found a middle level of knowledge about nourishment/nutrition. We think this level of knowledge should be raised and other factors determining healthy habits should be considered. It is necessary to go on with research and contextualize nourishment habits. PMID- 9741160 TI - [The health situation of the non-institutionalized elderly urban population of Castro Uridiales and the differentiating characteristics of those attending the health center frequently]. AB - OBJECTIVES: Main: to describe the health situation of the urban non institutionalised population of 75 or over in Castro Urdiales. Secondary. To analyse the differences in the over-user group. DESIGN: Descriptive, crossover. SETTING: Primary Care Health Centre. PATIENTS: People in the urban area aged 75 or over and with the following inclusion criteria: not institutionalised, who had been at least 6 months in the town, and for whom there was data for locating them. Over-users: people in the upper third of attendance (9 or more consultations per year). MEASUREMENTS AND MAIN RESULTS: A "Comprehensive Geriatric Assessment" was performed, cognitive state (Pfeiffer) and social assessment. The over-user group functioned better, had a better cognitive state and consumed more medication. CONCLUSIONS: The results coincided with other studies in most of the items analysed. It is important to be aware of the worse functional and cognitive condition of the group which attended the Health Centre least. Sub-groups of the elderly still need to be studied. PMID- 9741161 TI - [Passive smoking and other risk factors associated to the lower respiratory illnesses in sucking infants]. AB - OBJECTIVES: We try to evaluate the frequency of lower respiratory illnesses (LRIs), during the first year of life and it's relation to other risk factors, overall passive smoking and modes of feeding. DESIGN: This is a retrospective study in our city. SETTING: Primary Health Care. PATIENTS AND OTHER PARTICIPANTS: The study population is 240 children during the first year of life, born in 1993 and we have collected clinical information about risk factors and LRIs number. MEASUREMENTS AND MAIN RESULTS: A 37% have suffered from at least one episode of LRIs, during the first year of life. The parental smoking appears in 46.6%. The incidence of LRIs was strongly associated with passive smoking (OR = 1.86), exclusive breast-feeding and at least for 5 months (OR = 2.1) and older brothers (OR = 3.12). The number of episodes of LRIs was statistically significative higher in males (males 0.82 +/- 1.67 episodes; females 0.48 +/- 0.67 episodes) (p = 0.0489). CONCLUSIONS: In our report, the risk factors more strongly related with LRIs in suckling infants are passive smoking, older brothers, male sex and less than 5 months of breast feeding. PMID- 9741162 TI - [Prevalence of mixed anxiety-depression syndrome in those attending a care unit at a health center]. PMID- 9741163 TI - [The child behavior questionnaire: a useless tool]. PMID- 9741164 TI - [S-aminocaproic acid and hindrance to ejaculation]. PMID- 9741165 TI - [Feasibility of the COOP/WONCA vignettes and other instruments for Primary Care evaluation of health-related quality of life]. PMID- 9741166 TI - [Writing manuscripts]. PMID- 9741167 TI - [The evaluation of the hypertensive patient via self-measured blood pressure at home]. AB - OBJECTIVE: Evidence currently exist that treatment, and therefore prognosis, of the hypertensive patient can be improved by complementing the clinic blood pressure (BP) measurements with measurements taken out of the health care setting. The goal of the study was to validate a standard procedure of home BP self-measurement in contrast with a non invasive ambulatory BP monitoring (ABPM). METHOD: This was a transversal and descriptive study set at the primary care level. 58 hypertensive patients with poor office control were instructed to self measure their BP, with a previously verified digital sphygmomanometer, at home, 30 times for 10 days, and in the close 15 days an ABPM was taken for 24 hours. Reliability was determined comparing the degree of agreement between the different methods via the intraclass correlation coefficient (ICC) and the analysis of the individual differences. RESULTS: The self-measured BP and the ABPM do not differ significantly in systolic BP and only by 1.88 mmHg for the diastolic BP. The results taken in the office are significantly superior to those obtained by other methods -23 mmHg for the systolic BP and from 9 to 11 mmHg for the diastolic BP-. The ICC obtained comparing the results of the self-measured BP and ABPM are of 0.73 for systolic and 0.76 for the diastolic, whereas in the comparisons between office/self-measured, and office/ABPM the ICCs fluctuate from 0.23 to 0.50. The graphic analysis of individual differences allows the evaluation of the degree of reliability and the independence to the "white coat phenomenon". CONCLUSIONS: The BP self-measurement protocol that we present is a valid instrument, capable of confirming hypertension as well a suspicion of "white coat phenomenon" particularly when the availability of ABPM is scarce. PMID- 9741168 TI - [The effect of pregnancy on the consumption of tobacco and alcohol]. AB - OBJECTIVES: To determine if the knowledge of pregnancy modifies smoking and drinking habits and if some socio-demographic variables exert any influence in these changes. DESIGN: Descriptive observational study with a sectional design (prevalence study). SETTING: Obstetric service of a Spanish general hospital, and also three private centres in the same area. PARTICIPANTS: 271 women (mean age of 29 years) under 14 weeks of gestation (mean of 9 weeks) who attended to their first gynaecology visit and accepted to participate in the study. INTERVENTIONS: By means of direct interview to women, investigators filled a questionnaire with demographic characteristics, obstetric data, smoking and drinking habits and changes in these habits as a response to their pregnancy were also recorded. MEASUREMENTS AND MAIN RESULTS: Almost eighty percent of them planned their pregnancy, and 14.8% were using some contraceptive method when they became pregnant. Before learning of their pregnancy most of the women smoked cigarettes (53.5%), and a low proportion consumed alcohol at meals (20.3%) or between meals (10.7%). After knowledge of their pregnancy, 46.9% of smokers, 56.4% of drinkers during meals, and 86.2% of drinkers between meals, stopped these habits. Those women who smoked fewer cigarettes showed a higher capacity to stop consumption. It was also observed that those who gave up drinking at meals were those who drank less alcohol and who had no university degree. CONCLUSIONS: A high prevalence of smoking among fertile women was observed, although an important proportion of them stopped this consumption after learning about their pregnancy. This change was less frequent among the heavy smokers. Consequently, the establishment of specific programs for encouraging abstinence of alcohol and cigarette consumption in several population groups as described, could be recommended. PMID- 9741169 TI - [Indicators of drug provision and cost in the Avila health sector (1995)]. AB - OBJECTIVES: To analyse the prescription patterns in rural, urban and semi-urban Primary Care teams (PCT) and to study the variables which are correlated with the pharmaceutical cost, with special reference to medicines of low therapeutic value (LTV). DESIGN: Descriptive, crossover study. SETTING: Avila Health Area, 1995. PATIENTS AND OTHER PARTICIPANTS: Prescription profiles of 199 Primary Care doctors from Avila's 21 Health Districts, using the number of holders of the Individual Health Card (IHC) to determine population size. MAIN RESULTS: There were notable differences in prescription patterns between urban, rural and semi urban PCTs, with the "Drug cost per inhabitant per year" higher in rural than in urban PCTs (24,082 pesetas vs. 14,804), and the "Cost per prescription" less (1,417 vs. 1,577). In the simple regression analysis, there was a significant correlation between the variables "Number of LTV packages per inhabitant per year" and "Drug cost per inhabitant per year" (r = 0.68, 0.51 and 0.64 for urban, semi-urban and rural PCTs); between "Number of LTV packages per inhabitant per year" and "Number of prescriptions per inhabitant per year" (r = 0.79, 0.80 and 0.82); between "Drug cost per inhabitant per year" and "Number of prescriptions per inhabitant per year" (r = 0.81, 0.85 and 0.88); between "Number of IHCs per doctors" and "Drug cost per inhabitant per year" (r = -0.62, -0.57 and -0.31) and between "Number of IHCs per doctor" and "Number of prescriptions per inhabitant per year" (r = -0.51, -0.62 and -0.36). In the multiple regression analysis, if "Drug cost per inhabitant per year" was used as a dependent variable, the variable "Number of LTV packages per inhabitant per year" entered the model in urban and rural PCTs (Adjusted r2 = 0.55 and 0.60). If "Number of prescriptions per inhabitant per year" was used as dependent variable, the variable "Number of LTV packages per inhabitant per year" again explained, for all kinds of PCT, most of the variability (Adjusted r2 = 0.65, 0.65 and 0.76). CONCLUSIONS: The drug cost per inhabitant per year is less in urban than in rural Centres, while the cost per prescription is greater. The cost per inhabitant per year and the number of prescriptions per inhabitant per year have an inverse relationship to the population size. LTV medicines behave as pharmacy cost predictors, since a consistent relationship between their prescription, the cost per inhabitant per year and the number of prescriptions per inhabitant per year can be shown. PMID- 9741170 TI - [Consumption of medications in people over age of 65 years: potential problems and associated factors]. AB - OBJECTIVE: To determine the percentage of patients over 65 years of age with potential problems derived from the consumption of medications, and identify associated variables. DESIGN: A descriptive cross-sectional study. SETTING: Otero y Paulino Prieto Health Centers (Oviedo). PATIENTS: A randomly selected group of 298 patients over 65 years. MEASUREMENTS AND RESULTS: A questionnaire using both interview and clinical case notes provided data about sociodemographic details, perceived state of health, and functional status of patients, and qualitative and quantitative aspects of pharmacological treatment. 222 valid questionnaires were obtained. Once having identified patients with potential treatment problems, the associated variables were studied using bivariant and multivariant analysis. The mean consumption of medications was 3.1. 32.9% of patients had treatment problems (95% CI, 26.7-39.1), identified as drug interactions (54.8%), inappropriate use (36.9%) or both (8.3%). The bivariant analysis showed an association between the presence of problems and number of medications consumed (p < 0.001), prescribers (p < 0.001), diagnostic (p < 0.001), number of visit/year (p < 0.004), patient's own perception of health (p = 0.006), age (p = 0.018), socioeconomic status (p = 0.024) and cultural level (p = 0.039); the associations was not significative for functional status (p = 0.150), sex (p = 0.246), and number of UTB (Use Therapeutic Low) drug used (p = 0.751). Logistic regression showed an association between the presence of problems and patient's perceived state of health (OR, 1.56; 95% CI, 1.03-2.36) and the number of medications consumed (OR, 1.67; 95% CI, 1.41-2). CONCLUSIONS: A third of the population studied showed potential problems derived from the consumption of medications, through drug interactions and/or inappropriate use. Variables associated with this problems are the patient's perceived ill-health, and the highest consumption of medications. PMID- 9741171 TI - [Evaluation of anti-thrombosis treatment in patients with chronic non-valvular atrial fibrillation]. AB - OBJECTIVES: To assess both the indications of anti-thrombosis treatment in patients in our ambit with chronic non-valvular atrial fibrillation, and its observance in Primary Care. DESIGN: A descriptive, crossover, observational study of consecutive cases. SETTING: Third-level referral hospital in our Health District. PATIENTS: 132 adults first diagnosed with chronic atrial fibrillation between July 1st and December 31st 1996. MEASUREMENTS AND MAIN RESULTS: Patients' clinical records were used to assemble data on risk factors of embolism and counter-indications to prescribing antithrombosis treatment. A logistic regression model was performed to analyse the variables affecting the treatment at the time it was first given. 65 men (mean age 68.3) and 67 women (mean age 74.6) were included in the study. 87.9% of the patients had embolism risk factors; and 30.3% had at least one absolute or relative counter-indication to anti-coagulation. 79 patients had risk factors but no counter-indication, of whom 28% took anticoagulants, 39% had anti-aggregates prescribed and the remaining 33% received no anti-thrombosis treatment at all. Only 3 patients taking anticoagulants were referred to the Primary Care doctor. The regression model worked out signalled age under 75 and a previous embolism as factors associated with the indication and anticoagulants: We found no coherent regression model for the indication of anti-aggregates. CONCLUSIONS: Anti-thrombosis treatment is underused in Primary Care. An antecedent of an embolism is the most weight criterion for giving anticoagulants to patients. Age is shown to be the main reason for therapeutic reluctance to give anticoagulants to patients without counter-indications. There should be more patients being treated with anticoagulants in Primary Care. PMID- 9741173 TI - [Naturalistic studies to evaluate the effectiveness of medicines after being put on the market: why, when and how?]. PMID- 9741172 TI - [Rational use of non-steroidal anti-inflammatory drugs in primary care]. AB - OBJECTIVE: To quantify the consumption of non-steroidal anti-inflammatory drugs (NSAIDs) charged to the Andalusian Health Service and other health bodies, with a view to finding prescription quality and the pharmaceutical cost generated in the province of Sevilla in 1996. SETTING: The 759 pharmacies in the province of Seville. PARTICIPANTS: Beneficiaries of the Andalusian Health Service (pensioners and non-pensioners) and other health bodies. MEASUREMENTS AND RESULTS: The overall quantitative consumption indicators of the entire therapeutic group, and the dose per inhabitant per day (DID) of the systemic active principles which were significantly most prescribed, were determined. 3967487 NSAID prescriptions were dispensed, at a pharmaceutical cost of 2369, 483, 257 pesetas. Naproxen and Piroxicam were the active principles most used in terms of their DIDs. Of the population groups analysed, it is essentially the pensioners group which consumes these drugs. CONCLUSIONS: Seville province's NSAID use profile is inadequate, since the main drugs prescribed are those associated with high incidences of undesirable side-effects. PMID- 9741174 TI - [Scientific communication: author, editor, reviewer and reader]. PMID- 9741175 TI - [Commentaries on ecstasy]. PMID- 9741176 TI - [Transmission of the hepatitis C virus between cohabitants]. PMID- 9741177 TI - [Doppler examination in primary care]. PMID- 9741178 TI - [Oropharyngeal tumors of dogs--a clinical study of 79 cases]. AB - This study presents the data on incidence, TNM-classification and therapy outcome of 79 dogs with oropharyngeal tumors, which were admitted to the Clinic of Small Animals, Hannover School of Veterinary Medicine. 52 neoplasms were examined histologically. The most common tumors were malignant melanoma (n = 17), fibrosarcoma (n = 5), squamous cell carcinoma and peripheral odontogenic fibroma (n = 4). It could be determined that dogs treated by surgery, regardless of tumor type and type of surgery, had longer survival times than untreated dogs. With regard to survival time and the rate of local tumor recurrence, radical surgery (partial mandibulectomy/maxillectomy) led to good results in squamous cell carcinomas and invasive odontogenic tumors, but, keeping in mind the small number of cases, showed no advantage over conservative surgery in malignant melanomas, fibrosarcomas, neurofibrosarcomas and non invasive odontogenic tumors. It could be shown that the clinical staging of the patients was of prognostic relevance. PMID- 9741179 TI - [Continuing studies on the stability of sex steroids in the feces of cows over 12 weeks]. AB - In recent years an increasing number of reports were published indicating disturbances in the reproduction and sexual development in human beings and animals. These findings are seen in correlation with the increasing contamination of the environment with xenoestrogens. The importance of natural sexual steroids as a cause for this situation was poorly taken into account in the past. Therefore it was aimed by the study presented to contribute to knowledge in these questions. Samples of faeces were collected from 18 non pregnant and pregnant cows and their contents of estrogens and progesterone were analysed over a 12 weeks period as equivalent values of the standard substances estrone (E1) respectively 4-pregnene-20 beta-ol-3-one (20 beta P4) by using an enzyme immunoassay. The values of the substances of concern are decreasing gradually during the period of time examined. The median of the E1-equivalent dropped below the starting value in the ninth week, but the 20 beta P4-equivalent reached this level just in the second week. After the 12 weeks of observation under ambient temperature of 20 to 23 degrees C the examined equivalents of E1 and 20 beta P4 showed about 20% respectively 8% of the values measured at the beginning of the study. PMID- 9741180 TI - [Review of respiratory mechanics in animals. 3. Methodical and physiologic aspects of the use of the impulse oscilloresistometry system (OIS)]. AB - The impulse oscillometry system (IOS) which was originally developed for human medicine was found to be suitable for analysing respiratory mechanics in spontaneously breathing animals. This technique is non-invasive. METHODOLOGICAL ASPECTS: In order to use the IOS-technique in animals, a tightly fitting face mask is necessary. Furthermore, a flexible tube needs to be inserted into the measuring system. While the tube does not influence the measured results significantly, the face mask may affect the measurements. Therefore, its influence on the measured respiratory impedance must be taken into account. To prevent methodological errors, the head and body position of the animal should be standardised during the IOS-measurement. Since the methodological variability of the system is very small, the measuring results are highly reproducible. PHYSIOLOGICAL ASPECTS: In growing subjects, all parameters of respiratory mechanics depend on body weight. Since respiratory physiology is influenced by circadian rhythms, measurements of different days are only comparable when they have been done each day at the same hour. Comparing the respiratory impedance of different animals of similar age and body weight, a considerable inter-individual variability was observed. Due to the low level of physiological intra-individual variability, the method seems to be especially useful for studying influences on the respiratory system (i.e. pharmaceutics, therapeutics) in long-term studies using the same group of subjects. PMID- 9741181 TI - [Review of respiratory mechanics in animals. 4. The diagnostic affirmation ability of research using the impulse oscilloresistometry system (IOS) in calves]. AB - Taking methodological and physiological aspects into account (see Part 3), the impulse oscillometry system (IOS) was found to be sensitive to detecting and to quantifying clinically relevant changes in respiratory mechanics in calves. Therefore, the complex respiratory impedance needs to be measured in terms of resistance and reactance within the frequency range between 5 Hz and 20 Hz. The behaviour of resistance and reactance in dependence of frequency allows to differentiate and to localise airway obstructions. Obstructions of upper (extrathoracic) airways were mainly characterised by a frequency independent increase in the resistance. Within the reactance curve, no change in the resonant frequency could be observed. In a peripheral airway obstruction both resistance and reactance changed. The most typical finding concerning resistance was that a negative frequency dependence occurred. The reactance became more negative. Following this, the resonant frequency increased. With progressive obstruction of the peripheral airways, the reactance became more informative than resistance. PMID- 9741182 TI - [Improvement and acceleration of the diagnosis of contagious bovine pleuropneumonia by direct detection of the microbe using polymerase chain reaction (PCR)]. AB - A diagnostic procedure is described for the detection of Mycoplasma mycoides subsp. mycoides SC, the causative agent of contagious bovine pleuropneumonia. DNA extracted from clinical samples was investigated by the polymerase chain reaction (PCR) using at least 2 different primer pairs, one species-specific and another one specific for the class of Mollicutes. Using this method, the time required for detection of the pathogen was reduced to 2 days, whereas with traditional diagnostic methods (cultivation in broth, biochemical tests or immunofluorescence) the same finding would be available only within approximately 20 days. Although contagious bovine pleuropneumonia does not occur in Central Europe, there are occasional identifications of cattle having positive titres in the complement fixation test (CFT). Immunoblotting analysis of such sera confirmed that the reason for this phenomenon were cross-reactions with taxonomically related mycoplasma species. The present PCR assay proved to be suitable because of its rapidity, as well as high specificity and sensitivity. In the case of positive serological findings it enables diagnosticians to provide evidence on the presence or absence of the agent at short notice. PMID- 9741183 TI - [Are zoo Przewalski horses domesticated horses?]. AB - Analysed were the brain case capacities and brain weights of wild przewalski horses, przewalski horses from zoological gardens and domesticated horses. Domesticated horses have about 14% less brain case capacity and 16% less brain weight than wild przewalski horses. Przewalski horses from zoological gardens also have about 14% less brain capacity than wild przewalski horses. The brain weight of przewalski horses from zoological gardens shows no difference to the brain weight of domesticated horses. If we look at the brain size, przewalski horses from zoological gardens are domesticated horses. PMID- 9741184 TI - [Evaluation of occupational radiation exposure in veterinary radiographic diagnosis (a review)]. AB - A short review is given about fundamental dose units, the carcinogenic risk coefficients of ionizing radiation, the radiation exposure and potential radiation effects in the veterinary practice. The fundamentals of radiation protection comprise, inter alia, a modern and efficient X-ray and protection equipment, radiation safety training, radiation detection devices and, especially, restricting the size of the primary beam. PMID- 9741185 TI - [Pain therapy in dogs and cats--clinical experiences with buprenorphine (Temgesic)]. AB - Buprenorphine was used as a post-operative analgesic drug in 40 dogs and 30 cats. The analgesic properties of buprenorphine failed in 20% (dogs) respectively 23.3% (cats) of the cases even after high doses of 0.1 mg/kg in dogs and 0.01 mg/kg in cats. The unsatisfactory results were seen in fracture-patients with a concurrent severe tissue damage. Side effects even after high doses were of no clinical relevance. PMID- 9741186 TI - [The effectiveness of different disinfectants based on p-chloro-m-cresol against Ascaris suum eggs under laboratory conditions]. AB - The devitalizing effectiveness of 5 patterns of the disinfectant Neopredisan (active principle: p-chlorine-m-cresol) was tested to eggs of Ascaris suum in coincidence with the "guidelines for the testing of chemical disinfectants" of the German Veterinary Society (GVS). In the suspension-test all examined patterns indicated a 100% inhibition of development Ascaris suum-eggs. In the germ carrier test all patterns were in conformity with the necessities of the GVS-guidelines. Also in the modified testing method with a temperature in the refrigerator of +10 degrees C Neopredisan showed a reliable (100%) efficacy. The inhibition of development was clearly delayed with an admixture of 20% faeces to the suspension of eggs. In a suspension-test with Neopredisan 140 in 3% solution after 15 minutes with embryonated eggs a nearly 100% ovicide and larvicide respectively efficacy was proved. The investigations show, Neopredisan is of value as a suitable disinfectant in the complex of the combat against ascariasis in the breeding of pigs. PMID- 9741187 TI - [Blood chemical parameters for wild raptor patients and their changes after liver biopsy]. AB - The present paper tried to find relations between specific anamnesis of wild raptors and blood chemistry values at their day of presentation. 60 (88%) of 68 presented birds of prey showed changes in their blood values. In most birds an increase of GOT, GPT and AP was seen. Some birds showed increases of uric acid, urea and changes in the relation of Ca and P as well. A comparison between Eurasian buzzards with fractures and some without clinical signs showed a significant increase of uric acid, urea, potassium and inorganic phosphorus in the group of fractured birds. Changes of blood chemistry values after liver biopsy are investigated in the second part of the present study. Liver- and kidney values showed an increase after the biopsy. Kestrels (Falco tinnunculus) showed the maximum of the increase at the first day after biopsy while Eurasian buzzards (Buteo buteo) had the maximum at the third and Black kites (Milvus migrans) at the fifth day after biopsy. PMID- 9741188 TI - [Sensitivity of resonance thrombograms in thrombocytopenia of dogs]. AB - Based on results of 84 blood samples, taken from 28 dogs suffering from thrombocytopenia, the resonance thrombography turned out as a method with low sensitivity (S) to detect thrombocytopenia in dogs. It was insignificant which one of the two tested resonance thrombographs was taken for measurement and which parameter of the resonance thrombogram (RTG) was used for the thrombocytic potential of haemostasis, the amplitude (RTG-P) or descending time of the platelet side. Only samples containing < or = 25,000 platelets/microliter were reliably measured (S > or = 0.90), whereas thrombocytopenias with > 50,000 platelets/microliter usually resulted in false negative results. The correlation between the platelet count and RTG-P could be almost expressed by a geometrical regression (rs = -0.709). The low sensitivity of RTG mirrors the multifactorial influences and contrasts to the exclusive use of RTG in the screening of thrombocytopenia. PMID- 9741189 TI - [Some clinical symptoms and allergens on asthma-prurigo syndrome]. AB - The group of 146 patients suffering from asthma-prurigo syndrome (85 adults and 64 children) have been inquired in many various clinical centers. It was established that in 79.6% of the patients the first symptoms of illness appeared already in infancy and only 28.2% of the patients had negative familiar anamnesis on the allergy. In 73.2% of the patients with asthma-prurigo syndrome the symptoms of atopic dermatitis persisted longer than asthma symptoms and in 89.9% of them asthma-prurigo symptoms accompanied other form of allergic diseases. The most important causal allergens provoking asthma-prurigo symptoms were: house dust (in 64.4% of the patients), chocolate (in 42.2%), cat epithelia (in 40.2%) and cow milk proteins (in 29.5% of the patients). PMID- 9741190 TI - [Changeability of respiratory tract and skin symptoms in patients with asthma prurigo syndrome under the influence of emotional and environmental factors]. AB - In a group of 228 patients suffering from asthma-prurigo syndrome the influence of emotional state, permanent residence and season time on dynamic of the clinical symptoms were studied. The evaluation of emotional state in 80 adult patients was carried-out by means of Eysenck Personality Inventory. The emotional state had less negative influence on the exacerbation of the symptoms upon children then in adults (especially on their skin symptoms). It was confirmed that the patients with high level of neurotic symptoms revealed easier the exacerbation of asthma and skin disorders under the influence of emotional stress. The climatic treatment on the sea-side was more efficient for them than any mountain climatic cure, especially in the treatment of airways symptoms. Similarly, the summer season brought the relief of their symptoms from both the respiratory and skin symptoms. PMID- 9741191 TI - [Evaluation of thermovision images in pain syndrome associated with instability of the cervical segment of the spine]. AB - The aim of this work is to attempt to evaluate the characteristics of the thermovision image in pain syndrome associated with instability of the cervical segment of the spine (CSPS), to identify the variables and the impact of the characteristics of the thermovision image in the process of rehabilitation, and to specify the suitability of thermovision testing in the evaluation of rehabilitation. The results from tests performed in 71 patients indicate that patients with CSPS, in comparison to healthy subjects, are characterized by high asymmetry of the neck and severe cervical hyperthermia. When the characteristics of the thermovision image in the process of rehabilitation are traced, one can see a high dynamics of changes in these parameters, towards temperature symmetry in the neck and a reduction of cervical hyperthermia. The results of the tests point to the suitability of the thermovision testing in the evaluation of the effectiveness of rehabilitation procedures in patients with CSPS. PMID- 9741192 TI - [Mastopathy and simple goiter--mutual relationships]. AB - There have been assessed physical and ultrasonographic examination in patients with mastopathy and concentration of prolactin, thyrotropin, thyroxine in blood serum of these patients. The examination was carried out in 65 patients with mastopathy (group M), at the age ranging from 18 to 53 years, and in 30 healthy women (group K), at the age from 18 to 55 years as a reference group. The group of the patients with mastopathy and the control group were divided into three subgroups (taking into account the age and function of ovaries). There were excluded from the examined group patients whose general health state, particularly endocrinological disease, and applied drugs might condition occurrence of pathological changes in mammary glands. Particular attention was paid to exclusion from examination of patients with both primary and secondary hyperprolactinemia. Non-toxic goitre was found in 80% patients with mastopathy, and the results of palpation examination of thyroid were confirmed by thyroid ultrasonographic examination. Non-toxic goitre was significantly more often in patients with mastopathy in comparison with healthy women, and there was found significantly higher thyroid volume in these patients. The hormonal evaluation showed first of all significantly higher mean concentration of prolactin in blood serum of the patients with mastopathy than in the blood serum of healthy women in comparison with both the whole examined group and subgroups. There were not significant differences between the mean concentration of thyrotropic hormone, triiodothyronine and thyroxine in blood serum in premenopausal patients with mastopathy and mean concentration of these hormones in healthy women. Only postmenopausal patients were characterized by significantly lower mean concentration of triiodothyronine in comparison with the healthy subgroup. CONCLUSIONS: 1. In patients with mastopathy, there often coexists enlargement of thyroid gland, and prolactin may be also considered as an agent which influences genesis of a goitre. 2. It should be thus admitted that hormonal examinations with particular consideration to prolactin and thyroid hormones are appropriate management in diagnosing and treatment of patients with mastopathy. PMID- 9741193 TI - [Hypolipemic treatment in prevention of secondary ischemic heart disease]. AB - Available evidence indicating a possibility to inhibit the development of atherosclerosis and to reduce the mortality rate due to associated diseases in humans forms the rationale behind prevention of cardiovascular diseases. It consists of simultaneous modification of all concomitant risk factors including treatment of lipid disorders. Reduction of increased LDL cholesterol and TG levels is associated with a number of clinical benefits. It improves arterial endothelial function and stabilizes atheromatous plaque, which is reflected in the improved clinical, angiographic and hemostatic picture. Multiple studies have revealed a decreased rate of cardiac events and cardiovascular mortality in patients treated for lipid disorders. Furthermore, progression of atherosclerotic lesions has been shown to be inhibited. Before initiation of the therapy for lipid disorders taget LDL cholesterol level should be determined depending on the patient's risk group. In patients with known coronary artery disease or peripheral atherosclerosis (high risk group) it is desirable to reach LDL cholesterol level below 100 mg/dl and TG below 200 mg/dl. Basic approach to the therapy of lipid disorders both in primary and secondary prevention includes, modifications in the life style, especially dietary habits which may result in a mean of 10 to 20% decrease of LDL cholesterol and TG. When dietary modifications are ineffective it is recommended to use combined therapy but with a high degree of caution. PMID- 9741194 TI - [The role of estrogens in hormonal regulation of lipid metabolism in women]. AB - In the view of lipid metabolism, adipose tissue and liver are the most important tissues for 17-beta-estradiol, the main estrogen in women's body. The lack of estrogens in women after menopause may cause coronary heart disease. It is considered, that 25 to 50% of positive effect of estrogens which are given to postmenopausal women is connected with their action on lipid metabolism. Blood plasma parameters which characterize lipid metabolism return to their physiological values during estrogens therapy. Estrogens are transferred to adipose tissue cells and liver cells by endocrine and paracrine way. They are also produced in these cells from androgens. In adipocytes 17-beta-estradiol can be stored as its esters with long-chain fatty acids. It was proved that estrogens receptors are present in adipocytes and hepatocytes but their density is much lower than in gonads. On the cellular level estrogens regulate mRNA production for particular proteins among which there are proteins involved in lipid metabolism. In adipose tissue 17-beta-estradiol has a direct effect on lipoprotein lipase (LPL) and hormone-sensitive lipase (HSL). In the case of the first enzyme its synthesis is faster, while the synthesis of the latter is slower. On the other hand, indirect action of estrogens on adipose tissue is connected with the stimulation of the releasing of other hormones which increase HSL activity. To this group of hormones there belong catecholamines, growth hormone (GH) and glucagon. In liver 17-beta-estradiol regulates the rate of synthesis of structural apolipoproteins for VLDL and HDL. 17-beta-estradiol reduces the rate of apoB-100 synthesis, while stimulates apoA-I and apoA-II synthesis. HDL fraction containing apoA-I and apoA-II is necessary for chylomicrons and VLDL degradation as well as direct and indirect cholesterol transport to liver. Moreover, in hepatocytes estrogens stimulate the synthesis of apoC-III, while they decrease the synthesis of hepatic lipase (HL). In conclusion, 17-beta-estradiol by regulating lipid metabolism in adipocytes and hepatocytes modulates the concentration of lipid substances in plasma. The lack of 17-beta-estradiol leads likely to various lipid metabolism disorders in women after menopause. Estrogens therapy in these postmenopausal women may result in the improvement of lipid metabolism. PMID- 9741195 TI - [The influence of thyroid diseases and their treatment on the development of osteoporosis]. AB - Thyroid diseases and their treatment may influence osseous system. This work deals the problem of hyperthyreosis, hypothyreosis and thyroid hormones therapy with osteoporosis. There is reduced bone mass in hyperthyroid patients. Process of restoring lost bone matrix is not reached with euthyroid state, but lasts longer. Changes in bone mass in hypothyreosis are not of such clinical significance as in hyperthyreosis. Other problem is an influence of thyroid hormones therapy on mineral bone density. Data from literature dealing with this problem are divergent. Control thyroxin therapy with TSH level in normal range may diminish unprofitable influence on bone. Early diagnosis of osteoporosis has also substantial importance. Non-invasive methods play an important role. PMID- 9741196 TI - [Rhabdomyolysis: clinical features, causes, complications and treatment]. AB - Rhabdomyolysis is a condition affecting body homeostasis that results from impaired supply of muscles with energy, nutritional factors and blood. Complex pathophysiological mechanism causes that extended myolysis may complicate different clinical conditions, such as: crush syndrome, excessive physical effort (work, seizures), toxic effect of drugs and toxins, water-electrolyte disturbances, congenital enzymatic deficiencies etc. It seems that on the cellular level, essential role is played by excessively high intracytoplasmatic calcium level, which affects metabolic processes. So high calcium level is a consequence of muscular cell injury irrespective to its reason. It manifests clinically as muscular weakness, pal and oedema and laboratory tests reveal elevated CK, GOT, GPT, aldolase and LDH levels as well as dark brown urine colour. Demonstration of elevated serum myoglobin level or its presence in urine directly confirms development of rhabdomyolysis. In unfavorable conditions, rhabdomyolysis may result in acute renal failure. Appropriately early and adequate water supply and alkalization plays an essential role in prevention of impairment in renal function. In advanced phase of renal failure, hemodialysis is a standard treatment. PMID- 9741197 TI - [Causes of increased carbon dioxide partial pressure in arterial blood during general anesthesia]. AB - In healthy individuals, every physiologic and metabolic process is controlled by more or less complex manners. One of the best control mechanism is the way of excreting excess of CO2. The maintenance of artificial breathing compromises or completely disturbs the above process which exposes the body to an adverse effect of high paCO2. The possible causes leading to the increase in paCO2 during general anesthesia with the use of endotracheal intubation were discussed in the work. PMID- 9741198 TI - [Use of clonidine for perioperative therapy]. AB - Clonidine is a selective alpha 2 adrenergic receptors agonist with a wide spectrum of activity. Except well known hypotensive effect, clonidine stabilizes circulatory system and has sedative, anxiolytic, analgesic, diuretic etc. activities. Clonidine has some appliance during perioperative period. When used in premedication it has a lot of advantages: causes sedation, has anxiolytic properties, reduces secretion of saliva, stabilizes circulatory system, diminishes stress reaction, augments action of anaesthetic and analgesic drugs. When used during the operation, regulates circulatory system, prolongs and amplifies central and peripheral blocks. Clonidine diminishes patients requirement for opioids and local anaesthetics during postoperative and long-term pain therapy. PMID- 9741199 TI - [Hodgkin's disease in the thyroid gland]. AB - The paper presents one case with diagnosed the primary extra-nodular Hodgkin's disease in the thyroid gland. Patient was operated on. Hodgkin's disease was diagnosed postoperatively in the paraffin-embedded tissues. In authors' opinion progress in the imaging technics with fine-needle aspiration biopsy allows to formulate diagnosis, and treatment which includes chemo- and radiotherapy, improves survival. PMID- 9741200 TI - [Chromosome aberrations in patients with papillary thyroid cancer and other neoplasms]. AB - Cancer is essentially a genetic disease resulting from congenital or acquired alterations in some cells of the patient. Such changes may occur in particular oncogens and are responsible for the tumour phenotype of the affected population of cells. In contrast, unaltered tumour-suppressor genes are responsible for suppressing the neoplastic phenotype, and their inactivation by deletion or mutation permits cancerous development in the affected cells. The genetic model of carcinogenesis is based on the idea mutations at the DNA level, what creates a functional imbalance between the oncogenes and the tumour-suppressor genes, resulting in uncontrolled clonal proliferation. The ret/PTC oncogene is unique to papillary thyroid cancer. The paper presents a correlation analysis between chromosomal changes in papillary thyroid cancer and abnormalities of chromosomes in patients with breast cancer and chronic lymphocytic leukemia. PMID- 9741201 TI - [Sneddon syndrome--case report]. AB - We report a case of Sneddon syndrome in a 39-year-old woman who developed recurrent cerebral ischaemic events associated with livedo racemosa. We describe clinical and radiological features of this rare vasculopathy and laboratory findings essential for differential diagnosis. PMID- 9741202 TI - [Primary sclerosing cholangitis--tests with ubiquitin treatment. Case report]. AB - Treatment of chronic liver disease, including primary sclerosing cholangitis (CSP) is a difficult and still not fully solved problem. Both monotherapy and combined pharmacological therapy have shown little effect in inhibiting the disease process and preventing complications. The objectives of ubiquitin biotherapy were restitution of the immune system and inhibition of the disease process along with the stimulation of regenerative processes of the liver. In 1994 in Gastroenterological Clinic attempts at ubiquitin biotherapy were made with the use of thymus extract (TFX-JELFA ini.), which proved to be the ubiquitin preparation that was not only active in the immune system but also played a significant role in regenerative and adjuvant processes. It has been shown that ubiquitins play an essential role in proteolysis of proteins, their intracellular elimination and in the apoptosis. The early results of the clinical observations and laboratory tests indicate a gradual improvement and inhibition of the disease process the patient with CSP. PMID- 9741203 TI - [Aims and tasks of the Chambers of Physicians before the year 1939]. AB - The main aims of the Chambers of Physicians, which were called into being at the time when Poland was partitioned, were to protect the interest of physicians employed at health service after the introduction of compulsory insurance against sickness. They played a function of quasi-trade unions (in former Prussian sector). After Poland regained independence, based on the Chamber of Physicians Act of 1921 the Chief Chamber and regional chambers were created. Their aim was to assemble all doctors practicing in the country on obligatory basis. They performed a role of trade union, carried out social activity, ran saving and loan banks as well as funeral funds, controlled even distribution of physicians all over Poland, worked on ethical and deonthological codes, carried out arbitration by colleagues, performed the function of an organ of professional supervision. PMID- 9741204 TI - [Health personnel and viral hepatitis]. PMID- 9741205 TI - Serological markers of hepatitis A, B and C in first year student nurses. AB - OBJECTIVE: To know the prevalence of serological markers of hepatitis A, B and C virus in first year student nurses. SETTING: A transversal study of prevalence. SUBJECTS AND METHODS: 81 first year student nurses, mean age 20.6 years (18-37, S.D. 3.8), with demographic, epidemiologic and clinical variables, performing liver enzymes, anti-HAV IgG, anti-HBcore and anti-HCV. RESULTS: The anti-HAV IgG was positive in 9 students (11.1%), with a prevalence of 6.7% between 17 and 19 years (C.I.95% 1.7 to 19.3%), 8.7% between 20 and 22 years (C.I.95% 1.5 to 29.5%), 20% between 23 and 25 years (C.I.95% 10.5 to 70.1%), and 37.5% (C.I.95% 10.2 to 74.1) in those over 25 years (p < 0.001). No other significative variables existed according to age-group. Regarding hepatitis B virus, of the 65 not previously vaccinated, only 1 (1.5%, C.I.95% 0.08 to 9.4%) was immunized, and there were no cases of HBsAg positive. The anti-VHC was positive in one case (1.2%, C.I.95% 0.06 to 7.6%), RIBA indeterminate and with normal ALT. Only one student (1.2%) showed increased transaminase values, attributed to liver steatosis. None of the students had suffered any episode of acute clinic hepatitis. CONCLUSIONS: The anti-HAV IgG prevalence in first year student nurses in our area is very low, and it is not necessary to carry out prevacunal screening. The low prevalence of anti-Hbcore also rejects a similar screening with respect to HBV. There were no HBsAg or anti-HCV positive cases, but it should not be the cause of forgetting to take the universal precautions or giving a false sensation of security. PMID- 9741206 TI - Laparoscopic reconstruction of intestinal continuity following Hartmann's procedure. AB - Laparoscopy-guided reversal of Hartmann's procedure was performed in eleven patients who had been treated surgically for inflammatory disease or cancer of the colon. Restoration of intestinal continuity was achieved in ten of them. There were no postoperative complications. The mean surgical time was 144 minutes and the mean duration of postoperative ileus was 48 hours (range: 30 to 60 hours). The mean hospital stay was 7 days. Our results suggest that laparoscopic reversal of Hartmann's procedure is safer than and as effective as open surgery. PMID- 9741207 TI - Risk factors in the surgical management of perforated duodeno-pyloric ulcer. AB - The election of the treatment in the pyloric-duodenal perforations pursues a double objective: to reduce the operative mortality and to reduce the risk of ulcerous relapse in the long run. The authors carry out a retrospective study of 100 pyloric-duodenal perforations treated in 12 years. Three risk factors of immediate mortality are demonstrated: the age, 70 years or older; elapsed time, equal or superior to 24 hours and the existence of a situation of preoperative hemodynamic shock. The global mortality of the series was of 12% and the index of relapses of 12%. The authors outline a therapeutic management in which the election of the treatment was modulated for the factors of mortality risk. Thus, the presence of one of these factors must lead to the accomplishment of single treatment in treating only the perforation. The absence of risk factors must lead to accomplish a definitive treatment through the resection or the suture of the perforation followed by any type of vagotomy and eventually a drainage operation. The results obtained with Taylor's vagotomy, as well as with laparoscopic methods have not yet been validated although, probably, in a near future they will be integrated in the first management of pyloric-duodenal perforations. PMID- 9741208 TI - [Hepatitis B virus and the inflammatory/immunologic response (II): a new opportunity for the treatment of chronic hepatitis B and a suggestion for the treatment of other persistent viral diseases]. AB - With the immunologic rationale exposed in the first part of this paper, the authors analyze a new experimental treatment which includes the combination of an antiviral (ribavirin) and an immunomodulator (AM3) in a model of hepatotoxic viral-infection in mice. Rationale for this associated treatment is based on the ability of AM3 to restore the natural immunity through the induction of IL-12 and IFN-gamma. Furthermore, the treatment with AM3 decreases factor C3 of the complement system which appears to be implied in IL-12 downregulation. Experimental and clinical results showed herein suggest a new approach to the treatment of viral hepatitis which combines the use of antivirals in combination with immunomodulators able to restore the "immunologic chaos" induced by some viruses. PMID- 9741209 TI - [Cholestatic acute hepatitis induced by amoxycillin-clavulanic acid combination. Role of ursodeoxycholic acid in drug-induced cholestasis]. AB - We report two cases of acute hepatotoxicity after treatment with amoxicillin clavulanic. Viral hepatitis serology and autoantibodies were negative. Biliary tree obstruction and other etiologies were excluded. After discontinuation of the drug the evolution was favorable with clinical improvement and normalization of liver tests. Liver biopsy made in one patient showed cholestasic hepatitis with hepatocellular necrosis and other patient was treated with ursodeoxycholic. Also, we analyse potential utility of ursodeoxycholic acid administration in toxic cholestasis. PMID- 9741210 TI - [Epidermoid carcinoma in an anal fistula of long evolution]. PMID- 9741211 TI - [Salivary secretion in gastroesophageal reflux]. PMID- 9741212 TI - [Intense cholestasis associated with medicinal herbs]. PMID- 9741213 TI - [Acute ischemic proctocolitis]. PMID- 9741214 TI - Postmenopausal women: what is special. PMID- 9741215 TI - [The diagnostic and prognostic value of the 12-lead electrocardiogram in assessing the severity of coronary disease in the acute phase of an acute myocardial infarct]. AB - The conventional twelve-lead electrocardiogram (ECG) still is the cheapest, most used and absolutely essential diagnostic method for the acute phase of myocardial infarction (MI) allowing risk stratification and coronary prognostic evaluation in this phase mainly by the localization of the ST segment depression and/or T wave inversion (ST/T changes) not related to the infarct area in Q-Wave MI or at any localization in case of non-Q wave MI. The etiology and pathophysiology of these ST/T changes in the setting of MI has been controversial. With the objective of determining ECG prognostic and diagnostic value, 70 patients (Pts) (59 men and 11 women, mean age 58 + 13) admitted in the acute phase of MI were studied with revision of acute phase ECG ST/T changes. All patients underwent coronary angiography and ventriculography at the moment of hospital discharge. Patients were divided into two classifications: A) MI localization: A1--Q-wave MI (anterior--20 pts, inferior--29 pts, lateral--1 pt); A2--non-Q wave--20 pts. B) Evidence of ST/T changes outside the infarct area in Q wave MI or at any localization in non-Q wave MI (group B1--with ST/T changes, group B2--without ST/T changes). We correlated the angiographically documented coronary artery disease in groups with ST/T changes and their localization. RESULTS: A1) Anterior MI group: in the 6 pts (30%) with "opposite" (inferior) ST/T changes, right coronary artery (RCA) disease was documented in 5 and in the other 14 patients the RCA did not show significant lesions. Inferior MI group: in the 24 Pts (83%) with "opposite" (precordial) ST/T changes. 23 of them had angiographic correlation (left anterior descending (LAD) and/or circumflex (CX) artery disease). Lateral MI group: one Pt with anterior wall ST/T changes and LAD and CX disease. A2) Non-Q wave group: in 13 pts (87%) the diseased vessels were correlated with the site of ST/T changes. B1) Q-Wave AMI: left main and 3-vessel disease in 2 pts, 3-vessel disease in 17 pts, 2-vessel disease in 9 pts, 1-vessel disease in 2 pts and non-significant disease in one pt. Non-Q wave MI: left main and 3-vessel disease in 1 pt, 3-vessel disease in 7 patients, 2-vessel disease in 3 pts and 1-vessel disease in 4 pts. B2) non-Q Wave MI: 3-vessel disease in 5 pts, 2-vessel disease in 7 pts, 1-vessel disease in 6 pts and non-significant disease in 1 pt. Non-Q wave MI: 2-vessel disease in 2 pts and non-significant disease in 1 pt. IN CONCLUSION: When pts were divided according to MI localization, a correlation was found between the ST/T changes outside the infarct area with CAD in 91% of Pts in the Q-Wave infarction group, with more significance in inferior and lateral MI. In the non-Q wave group, we found correlation between the a coronary lesions and the localization of ST/T changes in 87% of the pts. The pt group with ST/T changes presented, when compared with the pt group without these changes, evidence of more severe coronary artery disease (CAD): 3 vessels or left main with 3 vessel disease. However, only in the Q-Wave infarction group was a statistically significant difference found between the group with ST/T changes compared to the group without these changes, concerning to the existence of more severe coronary disease. PMID- 9741216 TI - [Diastolic dysfunction in patients with chronic kidney failure on a hemodialysis program]. AB - OBJECTIVE: The aim of this study was to analyse different ultrasound parameters for the assessment of isolated left ventricular diastolic dysfunction (LVDD) in patients with chronic renal failure (CRF) on periodic hemodialysis (HD), comparing pulsed wave Doppler with pulsed tissue Doppler. MATERIALS AND METHODS: Forty-seven patients with CRF on HD (61% were male; mean age was 51.0 +/- 16.5 years, mean HD time--3.7 +/- 3.8 years, 38% had hypertension, 17% had diabetes) were studied by echocardiography (bidimensional, M-Mode, flow pulsed Doppler and tissue Doppler imaging). All patients had symptoms of left heart failure-class II NYHA, were in sinus rhythm and had no symptoms of ischemic heart disease. The presence of abnormal LV regional contractility was the exclusion criteria. According to their mitral inflow profile Doppler characteristics, patients were included in two groups: Group A (E/A > 1; n = 21) and B (E/A < 1; n = 26). We compared: LV dimensions and function, left atrial (LA) dimension. Gaasch index, LV mass index. E and A wave velocities (in flow pulsatile Doppler and tissue Doppler). E/N ratio in tissue Doppler, isovolumetric relaxation time (IVRT) and deceleration time (DT). RESULTS: There were no significant differences in the prevalence of age > or = 65 years male sex, hypertension or diabetes between group A and B patients, and almost all patients were on hemodialytic treatment for more than one year (81% vs 85%: NS). LV hypertrophy was present in almost all group A and B patients (A--95% vs B--85.5%; NS). Group A, compared with group B, had a difference in the Gaasch index (2.45 +/- 0.3 vs 2.08 +/- 0.4; p < 0.05), E wave velocity in flow pulsatile Doppler and tissue Doppler (cm/sec) (110 +/- 27 vs 62 +/- 20; p < 0.001 and 41 +/- 15 vs 28.5 +/- 16; p < 0.05), E/A ratio in tissue Doppler (1.3 +/- 0.4 vs 0.8 +/- 0.3; p < 0.001). IVRT (msec) (80.7 +/- 15.2 vs 113.5 +/- 28.3; p < 0.001) and DT (msec) (189.7 +/- 24 vs 278.2 +/- 17.9; p < 0.001). According to the E'/A' ratio in tissue Doppler, group A patients were divided in another two groups: E'/A' > 1 (13/21--62%) and < 1 (8/21--38%) and a significantly longer IVRT (75.8 +/- 9.3 vs 100.9 +/- 3.2; p < 0.001) and DT (178 +/- 15 vs 240 +/- 20; p < 0.001) and a greater LA dimension (37.6 +/- 6.9 vs 44.6 +/- 6.9; p < 0.05) were found. CONCLUSIONS: Pulsed wave Doppler is the most useful non invasive method for assessment of global diastolic dysfunction. In our study, 17% of the patients had E/A < 1 only in the tissue Doppler study. These patients probably had a pseudonormal mitral pattern. PMID- 9741217 TI - [Severe thrombocytopenia associated with Abciximab therapy--apropos a clinical case]. AB - The authors present the case of a 52-year-old male patient with severe coronary artery disease, submitted to percutaneous transluminal coronary angioplasty on a type C right coronary artery lesion. The use of a platelet glycoprotein receptor IIb-IIIa inhibitor, Abciximab, led to severe thrombocytopenia, treated only with platelet concentrate transfusions and with complete recovery. We also review the principal characteristics of the drug, its main indications and side effects. Severe thrombocytopenia has rarely been described in the literature. PMID- 9741218 TI - [Magnetic resonance in cardiology. The current clinical outlook]. AB - Magnetic resonance imaging (MRI) is a diagnostic technique used clinically in cardiology over the last ten years. It offers great advantages over other methods as it is noninvasive, does not use radiation, is tomographic and multiplanar. MRI is a very flexible technique based on the interaction between atomic nuclei, usually hydrogen, and external magnetic fields. This leads to image formation and to other important functional diagnostic information. MRI has undergone great technical developments and has proven to be most useful in several areas: cardiovascular structure; global and regional cardiac function; cardiovascular flow; coronary anatomy; myocardial perfusion and metabolism. The initial clinical diagnostic goals of MRI were mainly the anatomic characterization of aortic pathology, congenital heart disease, pericardium diseases and cardiac masses. With the development of cine MRI and, more recently, with rapid imaging and flow evaluation techniques, it has been increasingly used in the functional assessment of cardiovascular diseases. The functional study of the above mentioned clinical pathologies is now possible and information can also be obtained in other important areas; cardiac volumes and function; cardiovascular shunts, valvular heart disease; ischemic heart disease (ischemia detection, infarction quantification, coronary anatomy and flow). It also has an important role in myocardial characterization and is promising for myocardial metabolism, evaluated by spectroscopy. The goal of this review is the presentation of the main MRI techniques and the present clinical applications of this imaging method in cardiovascular diagnosis. PMID- 9741219 TI - [The renal dopaminergic system in chronic diseases of the renal parenchyma]. PMID- 9741220 TI - [Hodgkin's disease. An old disease with a new face]. PMID- 9741221 TI - [Persistent polyclonal B-cell lymphocytosis. A Western pattern versus and Oriental model?]. PMID- 9741222 TI - [Outcome of 70 patients diagnosed with Hodgkin's disease after first-line and salvage treatment: experience at one center]. AB - BACKGROUND: Retrospective analysis of 70 patients with Hodgkin's disease (HD) and treated consecutively in our center between 1979 and 1993. PATIENTS AND METHODS: All ganglionar biopsies were reviewed and finally 60 patients were selected: 43 males and 17 females with a median age of 39 years (12-79) and the following features: 55% III-IV stages, 25% extranodal involvement, 32% bulky mediastinum and 57% B-symptoms. Scheme of treatment: Only radiotherapy in 9 patients with localized stages and no adverse factors; only chemotherapy in 25 with advanced stages and combined therapy in 15 with localized stages and any adverse factor and in 7 with advanced stage and bulky disease. Chemotherapy was mainly based on MOPP until 1987 and MOPP/ABVD afterwards. Only four patients did not follow this scheme: 2 due to progression, 1 due to toxicity and 1 due to medical decision. RESULTS: Fifty patients achieved complete remission (CR), 2 partial and 8 minimal response or progression under therapy. Eleven patients have died: 6 due to HD, 4 of second neoplasias and 1 of an opportunistic infection. With a median follow-up for surviving patients of 4.8 years (1-2), the estimated overall survival at 8 years is 67.7% (SD 9%), tumor mortality 82.6% (SD 7%) and progression free survival 66.7% (SD 7%). Disease free survival for CR patients is 78.4% (SD 9%). Response to treatment was the only factor significantly associated with survival (p = 0.0001). The estimated survival for CR patients who relapsed is 92.3% versus 28% for those who did not obtain a CR (p = 0.0001). CONCLUSION: In our experience, first line therapy adjusted to clinical features at diagnosis has good results. Response to treatment has been the determinant factor for survival, mostly because salvage treatments, including high-dose chemotherapy approaches, have been of little effectiveness. PMID- 9741223 TI - [Prognostic factors in low-grade lymphoma]. AB - PURPOSE: Prognostic factors in low grade non-Hodgkin's lymphoma (LGL) are not well established. The aim of this study is to investigate prognostic factors on LGL treated in our institution during the last decade. PATIENTS AND METHODS: The study was carried out on 70 cases of newly diagnosed LGL, most treated with CVP or clorambucil and prednisone. The median follow-up was 37 months (1-132). Variables reported as prognostic factors in previous series were subjected to bivariate and multivariate analysis. RESULTS: Relevant clinical features were: Ann Arbor III-IV stage 74%, ECOG > or = 2-17%, bone marrow involvement 60% and large tumor burden according to MD Anderson criteria 21%. Complete response (CR) was achieved in 50% and partial response in 29%. In bivariate analysis factors related with poor CR were B symptoms, large tumor burden, high LDH and more than one extranodal site involvement. Logistic regression showed that large tumor burden (p = 0.02; OR = 0.07) and B symptoms (p = 0.07; OR = 0.14) were the best prognostic factors of poor CR. Five year global survival (GS) was 55%, with a median of 76 months. In univariate analysis factors related with GS were ECOG > or = 2, B symptoms, bulky, large tumor burden, retroperitoneo involvement and absence of CR. In multivariate analysis the only factor related with poor GS was large tumor burden (p < 0.00001; RR = 5.93). When therapeutic response was included in the model, absence of CR (p = 0.008; RR = 3.40) and large tumour burden (p = 0.005; RR = 3.86) were the factors selected. CONCLUSIONS: In LGL tumor burden was the most important prognostic variable. Tumor response showed less importance than in high grade lymphomas. PMID- 9741224 TI - [Efficacy of early erythropoietin use in critically ill, very-low-birthweight premature newborn infants: controlled clinical trial]. AB - OBJECTIVE: To determine the efficacy of erythropoietin in very low birth weight (VLBW) newborns less than 72 h of age. PATIENTS AND METHODS: We randomly assigned 40 critically ill newborn VLBW infants to receive either recombinant human erythropoietin (EPO) 150 units/kg per day (21 patients) or placebo (19 patients) during their first six weeks of life. The observers were unaware of the treatment assignments. Frequency of erythrocyte transfusion, adverse effects and haematologic measures were evaluated and compared. RESULTS: Before treatment gestational age, weight, haemoglobin, and pathology were similar in both groups. During the subsequent 6 weeks, haemoglobin and haematocrit of the placebo group fell significantly below those of the EPO recipients. More transfusions were received by the placebo recipients (7/21) than by the EPO recipients (2/21; p = 0.04). No adverse effects of EPO were observed. CONCLUSIONS: We recommend the administration of recombinant human erythropoietin since the first 72 h of age, because of the high frequency of anaemia, the efficacy of EPO and lack of side effects. PMID- 9741225 TI - [Report on the activity of the blood bank accreditation program (1987-1995)]. AB - PURPOSE: To show the incidence of the deficiencies detected in the Blood Banks for the accreditation by the Transfusion Accreditation Committee (CAT), previously named PABAS. MATERIALS AND METHODS: Analysis of the reports of the accreditation of 85 Blood Banks made by the PABAS during the period 1987-1995. RESULTS: Eighty-five (20.8%) of the 407 Community Blood Centers, Hospital-Based Blood Banks and Transfusional Services of Spain had been surveyed. There were 244 deficiencies, of which 31 (12.7%) were of the equipment, 114 (46.7%) of the procedures used, and 99 (40.6%) of the documentation. The activities with more incidence of faults were: Control of the temperatures of the storage of units 53 (21.7%), label of the components 38 (15.5%), quality system of the institution surveyed 32 (13.1%), transfusional procedures 30 (12.3%), and on the procedure of the selection of donors 29 (11.9%). By contrary, the areas of work with fewer incidences of faults were those related with the collection of the blood and components 10 (4.1%) and the laboratory 14 (5.7%). CONCLUSIONS: Low percentage of the Community Blood Centers, Hospital-Based Blood Banks and Transfusional Services, which ask to be accredited by the Transfusion Accreditation Committee. The 83.7% of the errors detected are of the procedures and documentation, which could be easily corrected by the training and continuous improving of the quality, and without need of new inversions in equipment. PMID- 9741226 TI - [Use of elements from the ISO 90000 system in Spanish hemotherapy]. AB - PURPOSE: To evaluate the prevalence of all twenty elements of ISO 9000 in the practice of the Blood Banks of Spain independently of being certified or not to ISO system. MATERIALS AND METHODS: By a survey sent in november 1996 to the Hospital-Based Blood Bank and Transfusional Services of 225 hospitals with more than 100 beds, and to 25 Community Blood Centers. The survey had 38 questions on the all elements of the ISO system and on other aspects of quality no directly related with it, as to be accredited by transfusional accreditation Committee, to have an hospital transfusion committee, informed consent of the transfusion and guidelines for using hemoderivatives. RESULTS: The survey was answered by 53 (21%) of hospitals and Community Blood Centers. None of the participant were certified to ISO system. The elements more used were the documentation of adverse reactions, procedures manual, validation of blood components, control of nonconforming products, and product identification and traceability, all of them used for more than 80% of participants. On the contrary, the elements less used were to have a quality unit, equipment manual, to validate the computer system, internal quality audits, criteria on purchasing, training and quality manual, all of them used by less than 30% of the participants. CONCLUSIONS: Generally speaking the elements of the ISO system more commonly used are those related to the basic ones of daily work and the least used are those related to the organisation aspects of quality. The Community Blood Centers score higher than hospital blood banks and among these, the best results are in the blood banks of hospitals more than 500 beds and the hospital accredited for teaching. PMID- 9741227 TI - [Changes of the complement system in myelodysplastic syndromes]. AB - PURPOSE: To search complement system activity and detection of circulating immune complexes in a group of patients with myelodysplastic syndromes (MDS) classified as with relatively favourable and unfavourable prognosis. PATIENTS AND METHODS: 28 patients with MDS were examined, 12 with refractory anaemia and 4 with refractory ringed sideroblastic anaemia, both subgroups of relatively favourable prognosis; 9 patients with refractory anaemia with blastic excess and 3 with refractory anaemia with blastic excess in transformation, subgroups of unfavourable prognosis. We determined factor B, alternate and classical hemolytic activity, as well as C3 and C4 quantification of the complement system. Circulating immune complexes detection was performed by the C1q deviation test and polyethylene glycol 6000 precipitation method at 3.75% final concentration. RESULTS: A significant decrease in C3 level, alternate haemolytic activity and factor B was demonstrated in the unfavourable prognosis subgroups in comparison with patients of favourable prognosis and normal controls. CONCLUSIONS: Our results suggest the possibility of subclinical infections in patients with unfavourable prognosis. PMID- 9741228 TI - [Immunosuppression induced by homologous blood transfusion]. PMID- 9741229 TI - [Treatment of Kasabach Merritt syndrome]. PMID- 9741230 TI - [Upper airway obstruction as a complication of acenocoumarol treatment]. AB - The case-histories of three patients treated with acenocumarol attended urgently because of sudden onset dyspnea and dysphonia an presented. Endoscopic and radiologic studies revealed in these patients a laryngeal obstruction by haematomas in a different grade, due to an excess in the effect of anticoagulant therapy, since prothrombine times measured as INR were too high. In one of the cases, the sudden and infrequent evolution associating initialy difficulty of breathing without any inspiratory or phonetic disorder, moved to a late diagnosis with a tragic ending, because it delayed an urgent tracheostomy, which bled profusely. It was a patient with a defective anticoagulant therapy control and numerous previous haemorrhagic incidents. The not so fulminant pattern in the other two cases allowed its early diagnostic and treatment planning, which included the admission to hospital and withdrawal of the anticoagulant drug and the vigilance of symptoms, up to confirm healing. Haemorrhagic laryngeal complications due to acenocoumarol, though unusual, imply an obvious vital trouble that moves to an urgent therapeutic attitude more or less aggressive. Its clinical suspicion obliges to its early discard. PMID- 9741231 TI - [Inflammatory pseudotumor of the spleen. An old concept with many questions]. AB - Inflammatory pseudotumor of spleen is an infrequent benign condition. It is difficult to differentiate, on a clinical and radiological basis, from haematologic neoplasms, granulomatous diseases as sarcoidosis and splenic hamartoma. Sometimes can be an incidental finding. Two women, aged 72 years, are presented. On the first case the sympthons mi micked a malignant disease. The second one was an incidental finding in a routine study for cholecystitis. Histological and immunohistochemical study showed a polymorphic cellular population including plasma cell, lymphoid cells, histiocytes, eosinophils and spindle cells, showing a reactive benign character. Plasma cells presented light chains polyclonality. Lymphoid cells were mature and with T inmunophenotype. Spindle cells were focally positive for muscle spe-cific actin and vimentine. In the first case, ultraestructural study showed myofibroblast morphology on the stromal spindle cells. Like many other authors have already postulated, immunohistochemical and ultraestructural findings would corroborate the mesenchymal reactive and benign nature of this type of lesions. PMID- 9741232 TI - [Streptokinase: correlation between different methods of biological evaluation]. AB - A study was carried out to establish an appropriate method for streptokinase (SK) potency determination (biological assay) in order to fulfil the main function of the Instituto Nacional de Medicamentos respecting products marketed in Argentina. The potency of different commercial samples of SK was determined against the International Standard, and three internationally accepted methods were used for this purpose: fibrin plate, clot lysis and chromogenic method. The analysis of results suggests that the fibrin plate method is the least precise and reproducible. The clot lysis and chromogenic methods demonstrated great precision and reproducibility, giving a correlation coefficient of 0.99. It is concluded that both of these methods are best suited to determine potency of SK commercial products. PMID- 9741233 TI - [Alveolar rhabdomyosarcoma with massive infiltration of the bone marrow as its initial manifestation]. AB - We report a case of alveolar rhabdomyosarcoma (AR) with massive infiltration of bone marrow at presentation, and initial diagnosis in bone marrow aspirate. A 35 year old man presented with a submandibular mass, and hematomas after mild traumatisms. Peripheral blood showed thrombocytopenia and a normocytic anaemia. Bone marrow film showed diffuse involvement by undifferentiated blasts with rhabdomyoblastic features. Subsequent biopsy of submandibular lymph node confirmed the diagnosis with positivity for specific muscle actin and desmin, and negativity for lymphoid markers. Initial presentation of AR with extensive bone marrow involvement is extremely rare, and it could lead to wrong diagnosis and treatment of acute leukaemia, with the serious consequences that this would have. Immunohistochemical study and morphologic differential features can be of great diagnostic help. PMID- 9741234 TI - [Ki-1+ large-cell anaplastic lymphoma with a leukemic appearance. Study of a case]. AB - Ki-1 anaplastic large cell lymphoma is a well-described subtype of non-Hodgkin's lymphoma with distinctive characteristics from the cytological, immunohistochemical and clinical points of view. One of the clinical behavior characteristic is that it rarely evolves into a leukaemic phase. We report the case of a 72-year-old man in which the appearance of tumor cells in peripheral blood was one of the most revealing information. The patient showed B-symptoms, bicytopenia and bone marrow involvement, together with hepatosplenomegaly and right axilar adenopathy, which after biopsied lead to Ki-1 anaplastic large cell lymphoma's diagnosis (stage IV-B). As far as the treatment and evolution are concerned, we choose a polychemotherapy (ACOP-B) because of the patient's age. Up to now clinical and analitical course is excellent and the patient is now in remission. PMID- 9741236 TI - [Testicular granulocytic sarcoma as a form of relapse in a patient with acute megakaryoblastic leukemia]. AB - Granulocytic sarcoma (GS) is a rare extramedullary tumor composed of myeloblasts and other granulocytic precursors. GS is mostly associated with myeloproliferative disorders, myelodysplastic syndromes and acute myeloid leukaemia. These tumors arise in the absence of leukaemia, at its initial diagnosis or at the time of recurrence. The most common sites of involvement are bone, skin, soft tissue and lymph node. Reports of GS in testis are very rare. We report an unusual case of GS in a patient with megakaryoblastic leukaemia arising in the left testis after four months in complete remission attained with low doses of Ara-C and granulomonocytic stimulating factor. PMID- 9741237 TI - [Genes, genomes and beyond]. PMID- 9741235 TI - [Primary MALT-type lymphoma of the breast]. AB - A 47 year old woman with a MALT lymphoma affecting the breast exclusively is reported. Complete response was achieved after CHOP chemotherapy but three years later a relapse was observed. Second-line chemotherapy (CNOP) and local radiotherapy were administered, and a second remission was obtained lasting for 1 year at last follow-up. A bibliographic search of MALT lymphomas of the breast showed that most such cases correspond to localized forms (stage I-II) and that from the radiologic point of view nodular lesions are almost the rule, although in the present case diffuse involvement was observed. MALT lymphomas of the breast tend to remain localized, and to relapse locally, and these facts appear as independent of the treatment applied (surgery, radiotherapy, chemotherapy or any combination). PMID- 9741238 TI - [Clinical impact of standardization in hematology. A vision through time]. PMID- 9741239 TI - [Sodium heparin and calcium heparin: units in which their potency should be expressed in Argentina]. PMID- 9741240 TI - [Quarantine and plasmapheresis]. PMID- 9741241 TI - [The Sebastian platelet syndrome: description of 2 cases and review of the literature]. PMID- 9741242 TI - [Budd-Chiari syndrome and primary myelofibrosis]. PMID- 9741243 TI - [Non-Hodgkin's lymphoma and von Recklinghausen disease in adults]. PMID- 9741244 TI - [Association of diabetes insipidus with a myelodysplastic syndrome prior to its transformation to acute leukemia]. PMID- 9741245 TI - [Diagnosis of postoperative deep venous thrombosis in hip replacement surgery: limitations of d-dimer]. PMID- 9741246 TI - [Refrigerated samples stored as whole blood can be valid for a deferred measurement of the INR]. PMID- 9741247 TI - [Amalgam--a question of belief? A review and assessment of the current literature]. AB - Several hundred written sources have been researched regarding old and new findings. Forgotten and new facts are covered in four chapters. One of the authors is a retired industrial chemist, and the other a dental surgeon. In the first section the natural and synthetic sources of mercury are explained. In every scientific study the ubiquity of mercury for billions of years must be taken into consideration. The second chapter looks at the toxicology of mercury and its compounds. Well known catastrophies in Japan and Iraq are depicted. Individual intoxications are hardly mentioned in literature. The third chapter is devoted to the dental amalgam, a silver alloy with both physical-chemical properties, and corrosive and thermal behaviour. An attempt is made to define the mercury balance. The forth paragraph deals with the disposal and recycling of mercury. Many dental associations are currently discussing the possibility of recycling amalgam, which is rather a stable silver alloy than a volatile mercury compound. A controlled landfill disposal of dental amalgam will absorb a certain quantity of mercury during its life cycle, and is positive. Recycling is expensive, energy consuming and causes additional exposure. Unless silver and mercury become rare metals, recycling should not be considered. As yet no worldwide regulatory procedures exist. PMID- 9741248 TI - [The rubber dam--the change in indications and technics]. PMID- 9741249 TI - [Complications when using bone cements]. PMID- 9741251 TI - [Bacterial infections as complications of dog bites]. AB - Dog bites may result in serious bacterial infections with e.g. the gram-negative rods Capnocytophaga canimorsus and Pasteurella multocida. Human disease caused by these microorganisms can be complicated by acute development of septicaemia and/or meningitis followed by disseminated intravascular coagulation syndrome, peripheral gangrene and renal failure. The mortality of C. canimorsus septicaemia is about 23-31%. These severe infections are most often reported in immunocompromised patients and occur a few days after the bite. By reviewing the literature it is concluded that the broadest prophylactic coverage is obtained by amoxicillin/clavulanic acid and that antibiotic prophylaxis should be given to all immunocompromised patients experiencing a dog bite. Moreover, prophylactic treatment should be initiated for all patients with greater penetrating wounds and those involving the hands. PMID- 9741250 TI - [Diagnostic value of C-reactive protein in bacterial infections. Review of the literature]. AB - Conflicting data for predictive values for C-reactive protein (CRP) in its ability to distinguish between viral and bacterial diseases are reviewed. Study designs regarding setting, patient-mix, severity of disease and prevalence seem to determine the magnitude of predictive values. We have calculated predictive values for patients suspected of septicaemia, meningitis, appendicitis, cholecystitis, upper- and lower respiratory disease, acute sinusitis and acute otitis media, and revealed the highest predictive values among patients suspected for severe and generalized infections. More localized diseases have lower predictive values. We emphasize the importance of a study design where the circumstances resemble the real situations in which the test is supposed to be used. This will ensure the clinical applicability of predictive values for a diagnostic test. PMID- 9741252 TI - [Occupational medicine in general practice. A study of the extent and nature of occupational injuries in the county of Ringkjobing]. AB - The purpose of this investigation was to estimate the occurrence and type of work related incidents in primary health care in the county of Ringkoebing, Denmark, with 270,000 inhabitants. Symptom-related general practitioner (GP)-consultations among 17-66 year-old residents were registered among 114 GP's during four days over one year. In each occasion the GP evaluated the question of work-relatedness versus none-work-relatedness. Of 3017 registered consultations, 479 were work related (15.9%). There were 376 disease cases and 103 injury cases. Musculoskeletal diseases constituted the dominating diagnostic category, as well as being the category with the highest proportion of work-relatedness, corresponding to 35% of all musculoskeletal consultations. Notification as a work injury case was performed for 19% of all work-related cases. This investigation represents the hitherto most extensive Danish material of the distribution of work-related incidents within the broad spectrum of the GP's practice. PMID- 9741253 TI - [Differences in the use of C-reactive protein analysis and erythrocyte sedimentation in general practice and hospitals. Development from 1986 to 1995]. AB - Analysis for C-reactive protein (CRP) was introduced in the Vejle Hospital catchment area in 1987. During the next ten year period the use of CRP has increased to reach a stable level both on hospital wards, out-patient clinics and in general practice. While the use of erythrocyte sedimentation rate on wards has decreased correspondingly, this is not seen in out-patient clinics or in general practice. There are medical as well as practical reasons for this discrepancy. It is believed that the possibility of performing a CRP as a point-of-care test in the GPs office in the future will lead to a reduction in the number of analyses of erythrocyte sedimentation rate performed in general practice as well. PMID- 9741254 TI - [Does a computerized price comparison module reduce prescribing costs in general practice?]. AB - The aim was to assess the trends in prescribed defined daily doses (DDD) and drug expenses before and after the introduction of a computerized cost containment module. On January 1993 the module was introduced in 20 practices in Aarhus County that used the computer record system APEX. Two control groups were made. From the Public Health Insurance in Aarhus County (600,000 inhabitants) data were collected during 1992 and 1993. Compared with te controls there were no changes in prescribed DDD, reimbursement for prescribed drugs, and reimbursement per prescribed DDD in the intervention group after the introduction of the module. Cost containment procedures should be more intensive than just giving the doctors a computer-assisted decision aid. PMID- 9741255 TI - [What do patients expect from the general practitioner? Danish results from a European study]. AB - This study aims to identify and explain which aspects of medical care patients valued and expected from the general practitioner. A questionnaire was given to a sample of 774 patients (86% response rate) from 15 practices. Patients rated the importance and ranked 40 different questions relating to their medical care. Aspects related to technical care were ranked highest. Patients gave low priority to aspects of care related to practice organisation. Enough time during consultations and quick services in case of emergencies got top rank in the study. The study provides information on patients' priorities regarding their general practitioner. PMID- 9741256 TI - [Osteoporosis in unselected patients in an ambulatory clinic for pulmonary medicine. An inexpensive diagnostic possibility not requiring extra time]. AB - Patients with pulmonary and allergy-related diseases often suffer from osteoporosis. Deformities of the thoracic spine in the lateral view of a routine chest X-ray were evaluated in 1558 patients admitted to a chest clinic in a prospective, blinded and non-selected manner with various cardio-respiratory complaints. Signs of osteoporosis were found in 108 of 811 males and in 79 of 747 females. In 187 patients with thoracic spine compression a total of 106 had received corticosteroid medication. Only one received oestrogen and no one took calcium or D-vitamin supplementation. Conclusively we found that more attention should be paid to osteoporosis in patients with pulmonary disorders. An easy and cheap way is to look for fractures on a simple chest X-ray in a population of patients with cardiorespiratory complaints. We found a very high prevalence of thoracic spine deformities, especially in men. PMID- 9741257 TI - [The significance of deletion polymorphism in the ACE gene for progression of diabetic nephropathies treated with ACE inhibitors]. AB - The aim of the study was to evaluate the effect of an insertion/deletion polymorphism of the angiotensin converting enzyme (ACE) gene on progression of diabetic nephropathy. We performed an observational follow-up study of 35 patients with insulin-dependent diabetes and diabetic nephropathy. Patients were investigated during captopril treatment for a median of seven (range three to nine) years. Eleven patients were homozygous for the deletion allele (DD) and 24 were hetero- or homozygous for the insertion allele (ID + II). The two groups had comparable glomerular filtration rate, albuminuria and blood pressure at baseline and captopril induced nearly the same mean reduction in blood pressure--to 103 (SD 5) mm Hg in the DD-group and 102 (8) mm Hg in the ID + II-group. The rate of decline in glomerular filtration rate was significantly steeper in the DD group than in the other group (mean 5.7 (SD 3.7) versus 2.6 (2.8) ml/min/year, p = 0.01). In conclusion, the deletion polymorphism in the ACE gene reduces the long term beneficial effect of ACE inhibition on the progression of diabetic nephropathy in patients with insulin dependent diabetes. PMID- 9741258 TI - [Urinary incontinence among patients admitted to a geriatric department. Prevalence and therapeutic needs]. AB - Urinary incontinence is a common condition in elderly people. It is well established that urinary incontinence in most cases can be significantly improved or cured by simple investigations and treatments. During a three month period 227 newly admitted geriatric patients were examined with the purpose of investigating 1) the prevalence of urinary incontinence; 2) the history of seeking medical treatment; 3) the present motivation for treatment; 4) the inconvenience the patients experienced from their incontinence. Fifty-seven percent of the women and 46% of the men suffered from urinary incontinence (mean age 81.8 years). Thirty-eight percent of the patients with present or previous incontinence had sought medical treatment. In 28% of these cases the treatment had been successful. The main reason for patients not seeking treatment was lack of expectation of treatment opportunities. Forty-four percent of the incontinent patients requested treatment in our geriatric department. Patients reported increasing inconvenience in line with increasing severity of the incontinence problem. Despite good treatment opportunities urinary incontinence in the elderly is still underdiagnosed and poorly treated. PMID- 9741259 TI - [Effect of an influenza vaccination campaign in the municipality of Copenhagen during the 1996-1997 season]. AB - In Denmark influenza vaccinations are usually paid for by the patients. In the autumn of 1996 Copenhagen City Council offered free influenza vaccinations to all residents aged 70 years or older. The impact of the campaign was evaluated in a questionnaire study of a random sample of the Danish population aged 70 years or older. In Copenhagen 81% (95% Confidence interval: 67-95%) of the elderly at risk were vaccinated compared to 51% (45-56%) outside Copenhagen. Offering free influenza vaccinations in a mass campaign is an effective way of improving coverage rate. However, no substantial difference was found in cost between the mass campaign and a targeted campaign with free vaccination in general practice. PMID- 9741260 TI - [Cardiac revascularization using both mammary arteries]. AB - Both internal mammary arteries in combination with veins were used for revascularization of the hearth in fifty Danish patients undergoing coronary artery bypass grafting (CABG) at Gentofte Hospital during the period 1994-1996. Patients with insulin-dependent diabetes mellitus, obesity, and age over 75 years were excluded. The patients were followed for at least one month after the operation. No patients died, and the complication rate was low and comparable to standard CABG using the left mammary artery and vein grafts. It is known from the literature that 10 years after CABG only 50% of vein grafts remain patent, and half of these have severe atherosclerosis. The mammary artery is far more resistant to atherosclerosis and 15 years after the procedure fewer patients have recurrent angina when both mammary arteries have been used. Bilateral mammary artery grafts can be used in half of CABG-procedures, and are especially indicated in younger patients. PMID- 9741261 TI - [Coronary bypass surgery using the radial artery. An alternative in patients with varicose veins]. AB - Arterial revascularization of the heart with the radial artery was performed in twenty patients with varicosities of the lower legs. The patients all had a good functional result and were free of angina pectoris after the operation. None of the patients had complications from harvest of the radial artery. The patients were mobilized early, as no veins had been harvested from the legs. PMID- 9741262 TI - [Unnecessary medical interventions. A questionnaire study among physicians and laymen]. AB - The aim of the study was to survey attitudes among medical doctors and lay people to unsolicited medical intervention. The design of the study was an anonymous questionnaire study including three scenarios implying ethical considerations. A total of 445 medical doctors working within different areas received the questionnaire, as did 75 medical students and 600 lay people. The results showed a response rate of 67%, highest among medical doctors and students. The lay people had significantly higher expectations concerning the medical intervention in two out of three scenarios compared to medical doctors and students. The participants were offered the opportunity to comment on the questionnaire. The conclusion of the Danish survey is that there is a significantly different approach to unsolicited medical intervention among lay people compared to medical doctors and students. Medical doctors are less disposed to perform unsolicited intervention compared with the wishes/expectations of the lay people. More open attitudes and information as well as better communication is recommended. PMID- 9741263 TI - [Heat of polymerization of bone cement can induce cardiac arrest]. AB - Total hip arthroplasty is associated with cardiopulmonary complications including cardiac arrest. We present one of four cases of cardiac arrest, two of the cases were fatal. The pathogenesis suggested to explain these complications is venous air embolism, generated by the methylmethacrylate bone cement polymerization causing thermal blood damage. To prevent this happening cortical bone allotransplantation around the prosthesis and bone cement with a low temperature of polymerization may be used. PMID- 9741264 TI - [Multiorgan failure and peripheral gangrene following a superficial dog bite]. AB - One case of life-threatening disease caused by the gram-negative rod Capnocytophaga canimorsus and a similar case without verification are presented. The severe diseases developed two to three days after a superficial dog bite and were characterized by acute development of septicaemia and fulminant disseminated intravascular coagulation syndrome. Moreover, the cases were complicated by renal failure, respiratory insufficiency and peripheral gangrene. PMID- 9741265 TI - [Primary pyogenic liver abscess in a child]. AB - A case report describing a child with a primary pyogenic liver abscess is presented. Primary pyogenic liver abscess is a rare disease at this age. Furthermore the clinical picture only rarely reveals indications of liver disease. To establish the correct diagnosis within the proper time, one must remember this diagnostic possibility. The recommended treatment is shortly discussed. PMID- 9741266 TI - [Brain natriuretic peptide. An aid in differential diagnosis of apnea]. PMID- 9741267 TI - [Crazy again!]. PMID- 9741268 TI - [Coenzyme Q10 and anticoagulants]. PMID- 9741269 TI - [Temperature measurement--how?]. PMID- 9741270 TI - [Mechanical ventilation in acute respiratory insufficiency in patients with chronic obstructive lung diseases]. AB - Chronic obstructive lung disease (COLD) is a common disease and cause of death. At an advanced stage, acute respiratory failure will repeatedly arise and mechanical ventilation may be the only solution. The best result of treatment is a return to the habitual condition. In this article a survey is presented regarding the course and survival following mechanical ventilation of acute exacerbation of COLD. Survival rate after mechanical ventilation was 50-70% compared to a survival rate of 75-95% in patients not mechanically ventilated. Mechanical ventilation influenced factors were taken into consideration. The severity of baseline COLD and comorbidity had significant influence on survival. These factors together with the quality of nursing were predictors of successful ventilator weaning. Knowledge of the patient before the need for mechanical ventilation arises is of decisive importance. PMID- 9741271 TI - [PNA--peptide nucleic acids. Towards gene therapeutic drugs]. AB - The principles of gene therapeutic "antigen" and "antisense" drugs are briefly presented. The target of such drugs is the genetic material of the cell--either the DNA itself, or its messenger molecules, mRNA. By exploiting base complementary principles, "antigen" and especially "antisense" drugs can easily be designed to target specific genes, the activity of which it is medically advantageous to inhibit. These could be virus- or oncogenes. In particular, the prospects of using peptide nucleic acids (PNA) as gene therapeutic drugs are discussed. PNA is a synthetic structural DNA mimic, which chemically is more closely related to peptides and proteins and which has so far shown very promising chemical and (molecular) biological properties in terms of its development into gene therapeutic drugs as well as diagnostic tools. PMID- 9741272 TI - [Evaluation of ear temperature measurements in a geriatric department]. AB - We evaluated an infrared tympanic thermometer (Genius 3000A) by comparing it with parallel measurements with an electronic rectal thermometer (Philips HP 5316) on 121 patients admitted to a geriatric department. Rectal temperature was on average 0.14 degree C +/- (ISD) above the ear temperature. 95% of the differences are within the interval from -1.18 degrees C to 1.46 degrees C. The coefficient of determination was only 0.30. The tympanic thermometer, Genius 3000A, cannot be recommended for daily use on a geriatric ward. PMID- 9741273 TI - [Induced abortion and risk of breast cancer]. AB - It has been hypothesized that an interrupted pregnancy might increase the risk of breast cancer, because proliferation of breast cells will take place without the protective effect of subsequent differentiation. In a cohort of 1.5 million women (28.5 million person-years) we identified 370,715 induced abortions in 280,965 women (2.7 million person-years) and 10,246 women with breast cancer. After adjustment for other risk factors, induced abortion was not associated with the risk of breast cancer (relative risk: 1.00; 95 percent confidence interval 0.94 to 1.06). However, the relative risk of breast cancer increased with increasing gestational age of the most recently induced abortion: < 7 weeks: 0.81; 7 to 8 weeks; 1.01; 9 to 10 weeks: 1 (reference); 11 to 12 weeks: 1.12; 13 to 14 weeks: 1.13; 15 to 18 weeks: 1.23; > 18 weeks: 1.89; P(trend) = 0.016. On a population basis, induced abortion was not associated with an increased risk of breast cancer. An increase was only seen for the special group of late second trimester abortions, but this finding was based on small numbers. PMID- 9741274 TI - [Peroperative radiologic findings of axillary lymph nodes in breast cancer]. AB - The series consists of 49 operable breast cancers, prospectively registered over a five month period. The removed axillary fat was peroperatively radiologically examined by trained radiologist, and the result was reported to the surgeon. Afterwards it was sent to the pathologist for a thorough histopathological examination. Peroperative radiographic examination showed that 30.6% of the resected axillary fat contained less than ten lymph nodes. Subsequent pathological examination found that only 8.2% actually contained less than ten lymph nodes. We conclude that peroperative radiological examination of removed axillary fat is not a reliable method to assess the number of lymph nodes removed during the surgical procedure. In our hands, a careful anatomical dissection removing all axillary fatty tissue and lymphatics including level I and II seems to be the method of choice. PMID- 9741275 TI - [Percutaneous fixation of sacral fractures. Primary experiences with a new technique]. AB - A number of techniques have been introduced to obtain reduction and fixation of rotational and vertical unstable sacral fractures and sacro-iliac (SI) joint disruptions. The purpose of this study is to report our primary experience with percutaneous cannulated screw fixation across the joint or fracture line. Fifteen consecutive patients, seven males and eight females, were operated. Six patients had isolated sacral fractures or SI joint disruption. All patients had a type C pelvic fracture according to Tile's classification. Percutaneous fixation of sacral fractures and sacro-iliac joint disruptions allows a short operating time, minimal bleeding and soft tissue damage, and immediate non-weight bearing mobilisation. No non-unions were seen and there were no cases of infection. In two patients the material had to be removed. The method gives adequate fixation of unstable posterior pelvic-ring fractures, but is technically difficult as malposition of the screws might cause serious perioperative complications. PMID- 9741276 TI - [Lumbar disk prolapse. Alcohol, tobacco and prognosis]. AB - The aim of the study was to examine whether smoking or the intake of different alcoholic beverages are associated with the outcome after first time lumbar disc surgery. One hundred and forty-eight patients consecutively operated upon for lumbar disk herniation over a one-year period were classified according to various social and demographic variables. Two and a half years later they were asked about their drinking and smoking habits and the outcome of the operation was assessed using a rating scale. Results showed that intake of wine, but not other alcoholic drinks, was associated with a good prognosis. Logistic regression analysis calculated that intake of wine was associated with a fourfold increase in success rate. This odds ratio was not significantly reduced by the following variables: Age, sex, smoking habits, employment status, social class, household income or marital status. In conclusion, intake of wine was associated with a good prognosis after lumbar discectomy. PMID- 9741277 TI - [Anterior interosseus nerve syndrome]. AB - A prolonged period of illness in a patient showing symptoms of reflexdystrophy is described. After going through several examinations and finally being referred to a pain clinic the patient demanded a second opinion from an orthopaedic surgeon. Her symptoms turned out to have been caused by a compression syndrome of the anterior interosseus nerve. PMID- 9741278 TI - [Glomus tumor of the temporal bone]. AB - A case story describing the typical symptoms and course of a glomus tumour of the temporal bone is presented. The most frequent symptoms are pulsatile tinnitus, unilateral hearing loss, aural fullness and paresis of the vagal nerve or other lower cranial nerves. The tumour is frequently visible by otoscopy. Despite being histologically benign, the tumour is infiltrative and may affect the surrounding cranial nerves or spread into the cranial cavity. The early signs and findings are vague. Since the sequelae are fewer when the tumour is treated while it is small, an increased awareness will be of benefit to the patients. PMID- 9741279 TI - [Multiple fractures following seizures in a pregnant woman]. AB - A 27-year-old woman, immigrated from Libanon, pregnant and at term was admitted with pain in the lower abdomen. The history was badly recognized because of the patient's inability to speak Danish. X-ray revealed a fracture of the left acetabulum with a minor central dislocation of the femoral head. During a new seizure she had another fracture of the right acetabulum also with a minor central dislocation of the femoral head and an undisplaced fracture of the proximal humerus on the left side. She was then delivered by a caesarean section. The seizures were later found to be due to grand mal type epileptic attacks. This case underlines the importance of a careful clinical and radiological investigation of patients with complaints of pain related to the central joints after seizures. PMID- 9741280 TI - [Traumatic knee luxation]. AB - This report is based on two case stories. Traumatic knee dislocation is a rare but serious event. There is extensive damage to the ligaments of the knee, but the vascular lesions, with an incidence of approximately 29% are of primary concern. Signs of ischaemia indicate arteriography or exploration, and if the vascular lesion is repaired within eight hours, the majority can avoid amputation. Complicating fractures, ligament lesions and nerve lesions have secondary priority to arterial lesions. A combination of surgical ligament repair/reconstruction and intensive rehabilitation seems the most promising. PMID- 9741281 TI - [Medial migration of a Hansson hook-pin after femoral neck fracture]. AB - Medial migration of a LIH (Lars Ingvar Hansson) hookpin is a very rare complication to internal fixation of hip fractures. We report a case of medial migration of an LIH hook-pin into the acetabulum. The importance of using the correct length hook-pin and ensuring that the base of the hook-pin is outside the lateral cortex of the femur is stressed. PMID- 9741282 TI - [Reflux symptoms--are trials with acid pump inhibitors useful in the diagnosis?]. PMID- 9741283 TI - [Imaging of the mouth]. PMID- 9741284 TI - [Melatonin, jet lag and the immune system]. PMID- 9741285 TI - [Neurosciences, psychiatry and psychoanalysis]. PMID- 9741286 TI - [On "prevention of hip fractures"--and common sense]. PMID- 9741287 TI - Contrast echocardiography as a new non-invasive diagnostic method for assessment of coronary artery disease: from endocardial border detection (EBD) to myocardial perfusion (MCE). PMID- 9741288 TI - [Local treatment of anal fissure with nitroglycerin ointment]. PMID- 9741289 TI - [Protect your patient against latex allergy]. PMID- 9741290 TI - [Tiaprofenic acid (Surgamyl) can cause severe chronic cystitis]. PMID- 9741291 TI - [Recall of slimming agents fenfluramine and dexfenfluramine--for what reason?]. PMID- 9741292 TI - [Grapefruit juice and drug interactions]. PMID- 9741293 TI - In vitro antigenotoxic activity of novel ginseng saponin metabolites formed by intestinal bacteria. AB - Ginseng saponin metabolites produced by human intestinal bacteria were evaluated for antigenotoxic properties by testing their effects on benzo[a]pyrene (B[a]P) induced mutagenicity and clastogenicity. They include 20-O-(beta-D glucopyranosyl)-20(S)-protopanaxadiol (IH-901), 20-O-(alpha-D-arabinopyranosyl(1- >6)-beta-D-glucopyranosyl]- 20(S)-protopanaxadiol (IH-902) and 20-O-[alpha-D arabinofuranosyl(1-->6)-beta-D-glucopyranosyl]-20(S)- protopanaxadiol (IH-903). IH-901, IH-902 and IH-903 inhibited the mutagenicity of B[a]P in a dose-dependent manner. In the chromosome aberration assay, IH-901 and IH-903 reduced the frequency of chromosome aberration induced by B[a]P. These results suggest that the ginseng saponin metabolites tested in the present study have potential as chemopreventive agents. PMID- 9741294 TI - In vivo effects of the kavapyrones (+)-dihydromethysticin and (+/-)-kavain on dopamine, 3,4-dihydroxyphenylacetic acid, serotonin and 5-hydroxyindoleacetic acid levels in striatal and cortical brain regions. AB - The in vivo effect of a single oral dose of 100 mg (+)-dihydromethysticin/kg body weight on striatal and cortical tissue concentrations of dopamine, serotonin, 3,4 dihydroxyphenylacetic acid and 5-hydroxyindoleacetic acid, as well as the dopamine and serotonin turnover, was tested in rats. Additionally, other rats were fed with a (+/-)-kavain containing food over a period of 78 days in order to calculate the influence of a chronic treatment with kavapyrones on the neurotransmitters. The results of the present in vivo study clearly demonstrate that neither (+)-dihydromethysticin in a high single dose, nor (+/-)-kavain chronically administered, altered the dopaminergic or serotonergic tissue levels in rats significantly. PMID- 9741295 TI - Prenylflavonoids: a new class of non-steroidal phytoestrogen (Part 1). Isolation of 8-isopentenylnaringenin and an initial study on its structure-activity relationship. AB - Bioassay-guided fractionation of a methanolic extract of a Thai crude drug, derived from heartwood of Anaxagorea luzonensis A. Gray (Annonaceae), resulted in the isolation of 8-isopentenylnaringenin (1) as an estrogen agonist with a activity of about an order of magnitude greater than genistein. Various flavonoids possessing isopentenyl side chains in the A-ring have been prepared and evaluated for their ability to bind estrogen receptor. In addition, enantiomers of 1 were separated and the respective enantiomers were assayed. These studies have demonstrated that the presence of an 8-isopentenyl group is an important factor for binding. Flavones, flavanones and flavonols having an isopentenyl substituent at C-8 exhibited an appreciable affinity for estrogen receptor. Conversely, isoflavones possessing an 8-isopentenyl substituent at C-8 did not show this activity. Movement of the isopentenyl group from position 8 to 6 resulted in loss of the activity. No significant difference was observed between 2(S)- and 2(R)-enantiomers of 1 in their binding affinity. Prenylflavonoids are reported to possess a wide range of biological activities; however, estrogenic activity has not been described. PMID- 9741296 TI - Prenylflavonoids: a new class of non-steroidal phytoestrogen (Part 2). Estrogenic effects of 8-isopentenylnaringenin on bone metabolism. AB - In order to examine whether 8-isopentenylnaringenin (1), which has been proven to possess estrogen agonist activity in in vitro tests, also produces in vivo estrogenic properties, the effects of 1 on uterus and on bone metabolism were determined in ovariectomized rats. Rats were ovariectomized and treated with 1 at 30 mg/kg/day subcutaneously for two weeks or 17 beta-estradiol at 0.01 mg/kg/day subcutaneously for two weeks. Ovariectomy resulted in an increase in urinary excretion of bone resorption markers (hydroxyproline, pyridinoline and deoxypyridinoline) and a decrease in bone mineral density of the proximal tibia as well as reduced uterine weight. Treatment with 1 or 17 beta-estradiol completely suppressed these ovariectomy-induced bone and uterine changes in a qualitatively similar manner. These results demonstrate that 1 acts as an estrogen agonist in the uterus as well as in bone in vivo. PMID- 9741297 TI - Flavan-3-ols isolated from some medicinal plants inhibiting COX-1 and COX-2 catalysed prostaglandin biosynthesis. AB - Extracts from the four plant species Atuna racemosa Raf. ssp. racemosa, Syzygium corynocarpum (A. Gray) C. Muell., Syzygium malaccense (L.) Merr. & Perry and Vantanea peruviana Macbr., traditionally used for inflammatory conditions, were fractionated using a cyclooxygenase-1 catalysed prostaglandin biosynthesis in vitro assay. The flavan-3-ol derivatives (+)-catechin, (+)-gallocatechin, 4'-O-Me ent-gallocatechin, ouratea-catechin and ouratea-proanthocynidin A were isolated as active principles. The IC50 values ranged from 3.3 microM to 138 microM whilst indomethacin under the same test conditions had an IC50 value of 1.1 microM. The flavonol rhamnosides mearnsitrin, myricitrin and quercitrin were also isolated. When further tested for inhibitory effect on cyclooxygenase-2 catalysed prostaglandin biosynthesis, the five flavan-3-ol derivatives exhibited from equal to weaker inhibitory potencies, as compared to their cyclooxygenase-1 inhibitory effects. The flavonol rhamnosides were inactive towards both enzymes. PMID- 9741298 TI - Inhibitory effects of Angelica pubescens f. biserrata on 5-lipoxygenase and cyclooxygenase. AB - Linoleic acid, osthol, osthenol and two polyacetylenes, falcarindiol and 11(S),16(R)-dihydroxyoctadeca-9Z,17-diene-12,14-diyn-1 -yl acetate were found to be the most active compounds responsible for the inhibitory activity of the dichloromethane extract of the roots of Angelica pubescens f. biserrata on 5 lipoxygenase (5-LO) and cyclooxygenase (COX-1) in vitro. They showed prominent inhibitory effect on 5-LO with IC50 values of 27.9 microM, 36.2 microM, 43.1 microM, 9.4 microM and 24.0 microM, respectively. Linoleic acid, osthenol, falcarindiol and 11(S), 16(R)-dihydroxyoctadeca-9Z,17-diene-12,14-diyn-1-yl acetate exhibited inhibitory activity on COX-1 with IC50 values of 13.3 microM, 64.3 microM, 66.0 microM and 73.3 microM. PMID- 9741299 TI - Anti-inflammatory activity of the alkaloid bukittinggine from Sapium baccatum. AB - The anti-inflammatory and related activities of bukittinggine were evaluated in comparison with reference drugs. The results obtained revealed that bukittinggine exhibited a significant inhibitory effect in carrageenin-induced hind paw edema and adjuvant-induced arthritis in rats. Its anti-inflammatory activity in both test models was comparable to that of acetylsalicylic acid. Bukittinggine also showed an inhibitory effect on the late proliferative phase of the inflammatory process in cotton pellet-induced granuloma formation in rats. In carrageenin induced rat pleurisy, bukittinggine exhibited marked inhibitory activity on exudative formation, accumulation of leukocytes and on PGE2-like activity in the exudate. Furthermore bukittinggine could significantly reduce fever in yeast induced hyperthermic rats and possessed analgesic activity comparable to that of acetylsalicylic acid when tested in acetic acid-induced writhing response in mice. However, bukittinggine exhibited only a weak effect in the tail-flick test when compared with morphine. It is likely that bukittinggine possesses a mechanism of anti-inflammatory, analgesic and antipyretic action similar to that of acetylsalicylic acid. PMID- 9741300 TI - Ilex aquifolium: protection against enzymatic and non-enzymatic lipid peroxidation. AB - The ethanolic extract of Ilex aquifolium L. (Aquifoliaceae) concentration dependently inhibited leukotriene B4 biosynthesis in isolated bovine PMNL with an IC50 value of about 60 micrograms/ml, whereas the effect on epidermal 12(S)-HETE biosynthesis was much less pronounced. The extract also inhibited the non enzymatic, peroxyl radical-stimulated lipid peroxidation in model membranes and was further a scavenger of the iron-dependent generation of hydroxyl radicals from hydrogen peroxide as determined by protection against deoxyribose degradation. While inhibition of leukotriene biosynthesis was not mediated by its known phenolic constituents such as hyperoside, rutoside, and chlorogenic acid, these compounds were responsible for the inhibitory effects of I. aquifolium against non-enzymatic lipid peroxidation and deoxyribose degradation. PMID- 9741301 TI - Inhibitory effect of Perilla frutescens and its phenolic constituents on cultured murine mesangial cell proliferation. AB - The inhibitory effects of Perilla frutescens and its phenolic constituents on cytokine-induced proliferation of murine cultured mesangial cells were investigated. DNA synthesis of mesangial cells stimulated by platelet derived growth factor (PDGF, 10 ng/ml) or tumor necrosis factor (TNF)-alpha (100 U/ml) was inhibited by the extract of P. frutescens (IC50 values, 3.3 and 1.4 micrograms/ml, respectively). The strength of the anti-proliferative activity was nearly equal in various chemotypes of P. frutescens. Caffeic acid, methyl caffeate, rosmarinic acid, and luteolin 7-O-glucuronide-6"-methyl ester were isolated as active constituents from the extract of the typical strain of P. frutescens, and their IC50 values for PDGF-induced mesangial cell proliferation were estimated as 26 microM, 2.6 microM, 1.8 microM, and 4.1 microM, respectively. We also compared the activities of related flavonoids previously isolated from P. frutescens, and luteolin had the highest anti-proliferative activity. PMID- 9741302 TI - Vitexicarpin, a flavonoid from the fruits of Vitex rotundifolia, inhibits mouse lymphocyte proliferation and growth of cell lines in vitro. AB - Certain flavonoids having a C-2,3-double bond were reported to show an inhibitory activity against T-lymphocyte proliferation, but not against B-lymphocyte proliferation in vitro. In the course of these studies, vitexicarpin (3',5 dihydroxy-3,4',6,7-tetramethoxyflavone) isolated from the fruits of Vitex rotundifolia was found to show potent inhibition against lymphocyte proliferation. Vitexicarpin inhibited T-lymphocyte proliferation as well as B lymphocyte proliferation at > 0.1 microM. IC50's were approximately 0.7 microM both for T- and B-cell proliferation. The inhibitory activity of vitexicarpin was reversible. Vitexicarpin also inhibited the growth of certain cancer cell lines, EL-4 and P815.9 (IC50 = 0.25-0.3 microM). These results suggest that vitexicarpin may be a potential therapeutic agent involved in inflammatory/immunoregulatory disorders such as rheumatoid arthritis and lymphomas. PMID- 9741303 TI - Conjugates of Tremella polysaccharides with microbeads and their TNF-stimulating activity. AB - A mannan (A) and a heteroglycan (D) were prepared by partial acidic hydrolysis of T3, a major polysaccharide fraction of the fungus Tremella fuciformis Berk. Methylation analysis of A and D showed that they contained a 1-->3 linked mannosyl main chain, to which were linked different side chains at position 2, 4, or 6 of the mannosyl residues. A and D were conjugated to albumin microbeads (AM) by reductive amination. The conjugates showed significant cytokine-stimulating activity in vitro whereas unconjugated AM had no activity. Not conjugated A and D showed cytokine-stimulating activity only in about 100 times higher concentrations. PMID- 9741304 TI - Antiprotozoal properties of 16,17-dihydrobrachycalyxolide from Vernonia brachycalyx. AB - Extracts of the leaves from Vernonia brachycalyx showed in vitro activity against Plasmodium falciparum and promastigotes of Leishmania major. The germacrane dilactone 16,17-dihydrobrachycalyxolide (1) which was previously isolated from the aerial parts of the plant was shown to be the major antiplasmodial principle. An X-ray crystallographic analysis established the absolute configuration and some signals in the NMR spectra were reassigned. 16,17-Dihydrobrachycalyxolide (1) elicited a strong antiplasmodial and antileishmanial activity but also a high toxicity against human lymphocytes. PMID- 9741305 TI - Desacetylmatricarin, an anti-allergic component from Taraxacum platycarpum. AB - The bioassay-guided fractionation of Taraxacum platycarpum (Compositae) extract led to the isolation of a desacetylmatricarin (1) as an active principle responsible for the anti-allergic property. It showed a potent inhibitory activity upon the beta-hexosaminidase release from RBL-2H3 cells in a dose dependent manner and the IC50 was 7.5 microM. Two structurally related guaianolide sesquiterpenes, achillin and leucodin, were also examined and their IC50 values were determined as 100 microM and 80 microM, respectively. PMID- 9741306 TI - The cytotoxic activity of a Salacia liana species from Monteverde, Costa Rica, is due to a high concentration of tingenone. PMID- 9741307 TI - Will the increasing prevalence of atopy have a favourable impact on rheumatoid arthritis? PMID- 9741308 TI - Ultrasonography in rheumatology: an evolving technique. PMID- 9741309 TI - Unusual and memorable. Erosive nodal osteoarthritis after frostbite. PMID- 9741310 TI - Mutual antagonism of rheumatoid arthritis and hay fever; a role for type 1/type 2 T cell balance. AB - OBJECTIVES: The balance between interferon gamma (IFN gamma) and interleukin 4 (IL4) producing T cells (T1 and T2 cells) seems to be of importance in many (auto)immune disorders. In general, T1 cell activity is important in cellular immunity whereas T2 cell activity plays a part in humoral responses. T1 cell activity predominates in joints of patients with rheumatoid arthritis (RA) whereas T2 cell activity is characteristic of atopic syndromes. This study investigated whether the prevalence of hay fever in RA is low and if severity of RA (T1 cell activity) can be influenced by the concomitant occurrence of a T2 cell mediated disease (hay fever). METHODS: The prevalence of hay fever was assessed in 643 consecutive (RA and non-RA) patients seen in our outpatient clinic and confirmed by skin test and specific IgE. Of this group the 12 RA patients with hay fever were compared with RA patients without hay fever (matched for age, sex, and disease duration). RESULTS: The prevalence of hay fever in RA patients is lower than in non-RA patients (4% versus 8%), and yields a relative risk for RA patients to develop hay fever of 0.48. RA patients with hay fever showed a lower disease activity (erythrocyte sedimentation rate, C reactive protein, Thompson joint score, and radiographic joint damage (Sharp) score) than RA patients without hay fever. The clinical data were related to peripheral blood T1/T2 cell balance: a lower IFN gamma/IL4 ratio was observed for RA patients with hay fever, indicating a comparatively increased T2 cell activity in RA patients with hay fever. CONCLUSION: These results argue in favour of the exploration of treatments aimed at regulation of a possible imbalance in T1/T2 cell activity in RA. PMID- 9741311 TI - Serum androgen-anabolic hormones and the risk of rheumatoid arthritis. AB - OBJECTIVE: It has been hypothesised, mainly on the basis of indirect evidence, that low serum concentrations of androgen-anabolic hormones would play a causal part in the aetiology of rheumatoid arthritis (RA). METHODS: A case-control study was nested with a Finnish cohort of 19,072 adults who had neither arthritis nor a history of it at the baseline examination during 1973-1977. Pre-illness serum specimens for the assay of testosterone and dehydroepiandrosterone sulphate (DHEAS) were available from 116 cases who had developed RA by late 1989. Three controls per each incident case were individually matched for sex, age, and municipality. RESULTS: The mean testosterone concentration was 1.4 nmol/l in those 84 women who developed RA and 1.4 nmol/l in their controls; the corresponding figures for DHEAS were 5.2 mumol/l and 5.5 mumol/l, respectively. Mean testosterone concentration in the 32 male cases was 26.1 nmol/l and 26.4 nmol/l in their controls; the corresponding figures for DHEAS were 11.2 mumol/l and 10.1 mumol/l, respectively. Analysis by subgroups (rheumatoid factor positive and negative disease, pre-menopausal and postmenopausal women) and by hormone distributions showed no differences. CONCLUSION: The findings are not in line with the contention that low concentrations of testosterone and DHEAS play a part in the aetiology of RA. PMID- 9741312 TI - Influence of HLA-class II incompatibility between mother and fetus on the development and course of rheumatoid arthritis of the mother. AB - OBJECTIVE: To assess the relation between the course of rheumatoid arthritis (RA) during pregnancy or the onset of RA postpartum and DRB1, DQA1, and DQB1 incompatibilities between mother and child. METHODS: In 45 pregnancies of 33 RA patients the course of RA was related to the number of class II incompatibilities. Furthermore class II incompatibilities in 16 pregnancies followed by RA onset were compared with those in 87 control pregnancies. RESULTS: The risk of a favourable compared with an unfavourable course was 0.95, 2.67, and 2.38 in case of DRB1, DQA1, and DQB1 incompatibility respectively. DQA1 and DQB1 incompatibilities were seen more often in the 10 pregnancies followed by RA onset within three months than in control pregnancies (OR 8.02, 95% CI 0.97, 66.06 and OR 8.79 95% CI 1.07, 72.46 respectively). CONCLUSIONS: DQA1 and DQB1 incompatibility between mother and child seems to have a favourable effect on the course of RA and may postpone the risk of RA onset during pregnancy. PMID- 9741313 TI - Fatigue in primary Sjogren's syndrome. AB - OBJECTIVE: To assess fatigue in relation to depression, blood pressure, and plasma catecholamines in patients with primary Sjogren's syndrome (SS), in comparison with healthy controls and patients with rheumatoid arthritis. METHODS: For the assessment of fatigue the Multidimensional Fatigue Inventory (MFI) was used, a 20 item questionnaire, covering the following dimensions: general fatigue, physical fatigue, mental fatigue, reduced motivation, and reduced activity. Furthermore, the Zung depression scale was used to quantify aspects of depression. Forty nine female primary SS patients, 44 female patients with rheumatoid arthritis (RA), and 32 healthy women filled in both questionnaires. In addition, supine values of blood pressure and plasma catecholamines were measured in the patients with primary SS. RESULTS: Primary SS patients were more fatigued compared with the healthy controls on all the five dimensions of the MFI. When the analyses were repeated using depression as a covariate, group differences disappeared for the dimensions of reduced motivation and mental fatigue. In the primary SS patients, significant positive correlations between depression and the dimensions of reduced motivation and mental fatigue were found. Comparing patients with primary SS with those with RA, using depression as covariate, no statistically significant differences were found between these groups. No relation between fatigue and blood pressure was found, but a negative correlation was observed between the general fatigue subscale of the MFI and plasma noradrenaline. CONCLUSION: Patients with primary SS report more fatigue than healthy controls on all the dimensions of the MFI and when controlling for depression significant differences remain on the dimensions of general fatigue, physical fatigue, and reduced activity. The negative correlations between levels of noradrenaline and general fatigue in patients with primary SS may imply the involvement of the autonomic nervous system in chronic fatigue. PMID- 9741314 TI - Heart involvement in systemic sclerosis: an ultrasonic tissue characterisation study. AB - BACKGROUND: Clinicoepidemiological findings indicate that symptomatic heart involvement in patients with systemic sclerosis (SSc) predicts a very poor prognosis. At necropsy studies, SSc heart involvement without significant coronary lesions is characterised by patchy myocyte necrosis and contraction band necrosis with collagen replacement leading to myocardial fibrosis. There is a discrepancy between the frequency of clinically evident myocardial disease (25%) and autoptical myocardial fibrosis (81%). OBJECTIVE: The aim of this study was to detect preclinical myocardial alterations in SSc patients by ultrasonic videodensitometric analysis. METHODS: Thirty five SSc patients (three male, aged 48.6 (11) SD years, range 22-65) with normal ventricular function and 25 age and sex matched healthy controls were studied. All patients had a negative maximal exercise stress; in all cases arterial hypertension, renal involvement, and diabetes were excluded. Echocardiographic images were digitised by a real time videodigitiser (Tomtec Imaging Systems). Quantitative texture analysis was performed on data from the septum and the posterior wall, obtaining mean gray level histogram (MGL) at both end-diastole (d) and end-systole (s). The cyclic variation index (CVI), was calculated according to the formula ((MGLd-MGLs)/MGLd) x 100. Left ventricular mass (LVM), body surface corrected, was calculated according to Penn convention. RESULTS: Comparable systolic and diastolic blood pressure, LVM, diastolic and systolic function were recorded in both SSc patients and controls. In contrast, in SSc patients the CVI, which is the expression of the intrinsic myocardial structural function, was significantly lower than in controls (septum: -18 (28)% v 35 (10)%, p < 0.0001; and posterior wall: -13 (32)% v 50 (20)%, p < 0.0001). Changes in cyclic echo amplitude, probably related to myocardial fibrosis, were detected in the large majority of SSc patients (88%). CONCLUSIONS: Ultrasonic videodensitometric analysis represents a non-invasive, feasible method that can detect early myocardial changes in SSc patients, which could be related to both fibrosis and microcirculatory abnormalities. Their potential evolution towards ventricular dysfunction and their link with cardiac sudden death, because of severe conduction system or rhythm disturbances, should be further investigated. PMID- 9741315 TI - Expression of alpha and beta subunits of the integrin superfamily in articular cartilage from macroscopically normal and osteoarthritic human femoral heads. AB - OBJECTIVE: The objective of this study was to detail the topographical and zonal distribution of alpha and beta subunits of the integrin superfamily in normal and osteoarthritic cartilage. METHODS: Immunohistochemistry utilising antibodies towards alpha and beta subunits was performed on cryostat sections of human articular cartilage from macroscopically normal (n = 6) and osteoarthritic (n = 6) femoral heads. Samples of articular cartilage were obtained from 12 topographically distinct sites from each femoral head. Each section was divided into zones (superficial, middle, deep) and staining scores were recorded. RESULTS: Normal cartilage stained for integrin subunits alpha 1, alpha 5, alpha V, beta 1, beta 4, and beta 5, but not for alpha 2, alpha 3, alpha 4, alpha 6, beta 2, beta 3, and beta 6. Intact and non-intact residual cartilage from osteoarthritic femoral heads stained for alpha 1, alpha 2, alpha 5, alpha V, beta 1, beta 4, and beta 5. Staining was occasionally seen for alpha 4 and beta 2, but not for alpha 3, alpha 6, beta 3, and beta 6. There was no topographical variation in the staining for any of the subunits in either normal or osteoarthritic cartilage. The only subunit that displayed a zonal variation was alpha V; staining for this subunit was most pronounced in the superficial zone compared with the middle and deep zones. CONCLUSION: Chondrocytes in normal and osteoarthritic cartilage express the integrin subunits alpha 1, alpha 5, alpha V, beta 1, beta 4, and beta 5. Chondrocytes in osteoarthritic cartilage, in addition, express the alpha 2, alpha 4, and beta 2 subunits. The alpha v subunit is expressed by more chondrocytes in the superficial zone in comparison with cells in the deeper zones. None of the subunits display topographical variation in expression. PMID- 9741316 TI - Iron, lactoferrin and iron regulatory protein activity in the synovium; relative importance of iron loading and the inflammatory response. AB - OBJECTIVES: To determine the ability of lactoferrin in rheumatoid arthritis (RA) synovial fluid to bind "free" iron, and to study the regulatory mechanisms therein that control iron homeostasis. METHODS: "Free" iron was determined by the bleomycin assay and lactoferrin concentrations by enzyme linked immunosorbent assay. The activities of iron regulatory protein (IRP) and NF-kappa B in synovial fluid cells were assayed by mobility shift assay. RESULTS: 30% of synovial fluids contained "free" iron and in these, lactoferrin concentrations were significantly lower than in those with no "free" iron (p < 0.01). Addition of exogenous lactoferrin consistently reduced the amount of "free" iron in positive synovial fluids. IRP activity in synovial cells did not correlate with synovial fluid iron concentrations but did correlate with NF-kappa B activation and with serum C reactive protein. CONCLUSION: Lactoferrin may prevent iron mediated tissue damage in RA by reducing "free" synovial iron concentration when inflammatory stimuli have disregulated IRP mediated iron homeostasis. PMID- 9741317 TI - Prevalence of rheumatoid arthritis in Italy: the Chiavari Study. AB - OBJECTIVE: To ascertain the prevalence of rheumatoid arthritis (RA) in an Italian general population. METHODS: The study was performed in the years 1991-92 in Chiavari, a small town located on the Ligurian coast, and involved 4456 subjects aged 16 years or more from four general practices. The subjects received a postal questionnaire developed to detect patients with current or past inflammatory joint diseases. The age and sex distribution of the sample were similar to those of the Italian population from the 1992 census. Patients reporting a history of joint swelling in at least a pair of symmetrical joints were reviewed by a rheumatologist. The clinical records of non-responders and responders who failed to attend the clinic were also reviewed. RESULTS: 3294 of 4456 (73.9%) subjects answered to the questionnaire. The mean (SD) age of the 3294 responders was 48.3 (19.3) years; 53.7% of them were female. Swelling in at least two symmetrical joints was reported by 230 subjects (7%). Among them, 11 patients fulfilling the 1987 ARA criteria for RA were identified. The prevalence of RA was 0.33% (95% CI 0.13, 0.53) in the general population, 0.13% (95% CI 0, 0.31) in men, and 0.51% (95% CI 0.18, 0.84) in women. CONCLUSIONS: These data are consistent with the results of three earlier studies published in the fifties in the Italian literature and confirm that the prevalence of RA is low in Italy and has remained unchanged in the last 40 years. PMID- 9741318 TI - TCR beta spectratyping in RA: evidence of clonal expansions in peripheral blood lymphocytes. AB - OBJECTIVE: To compare the TCR beta repertoire of peripheral blood CD8 enriched (CD8+) and depleted (CD8-) T cells in rheumatoid arthritis (RA) patients and controls using CDR3 length analysis (spectratyping). METHODS: CD8+ and CD8- T cells were separated from 14 RA patients and 12 controls, using magnetic beads coated with anti-CD8 monoclonal antibodies. cDNA was prepared as the template for amplification with 22 V beta-C beta primer pairs. The products were resolved by electrophoresis in an ABI373 sequencer using GENESCAN software. Expansions were identified as dominant CDR3 lengths, where the area underlying the corresponding peak exceeded the sum of the areas of the two adjacent peaks. This method was validated by sequencing 10 samples displaying dominant peaks. The expansion frequencies in RA patients and controls were compared using the chi 2 test statistic. RESULTS: Dominant peaks were evident in several V beta families. They were more frequent in RA patients in both the CD8+ subset (RA normalised frequency 10.6; control normalised frequency 8.0; p = 0.03) and the CD8- subset (RA normalised frequency 2.9; control normalised frequency 1.5; p = 0.02). Sequencing of 10 samples exhibiting dominant peaks revealed an unequivocal clonal expansion in nine (90%). CONCLUSIONS: RA patients exhibited a significantly increased frequency of T cell expansions both in the CD8+ and CD8- subsets. This phenomenon may reflect the proliferation of autoreactive cells, a nonspecific expansion of memory T cells in response to pro-inflammatory cytokines or a defect of T cell regulation that predates the onset of RA and may itself predipose to disease. PMID- 9741319 TI - Protective effect of gold rings and rheumatoid arthritis. PMID- 9741320 TI - Gold and ring finger. PMID- 9741321 TI - Rheumatology outpatient training. PMID- 9741322 TI - Cancer-induced bone diseases. PMID- 9741323 TI - Bone metastases--the clinical problem. PMID- 9741324 TI - Mechanisms of tumour metastasis. PMID- 9741325 TI - Parathyroid hormone-related protein (PTHrP) and hypercalcaemia. PMID- 9741326 TI - Morphology of bone metastasis. PMID- 9741327 TI - Cellular and molecular mechanisms of breast and prostate cancer metastasis to bone. PMID- 9741328 TI - Myeloma bone disease. PMID- 9741329 TI - Monitoring of bone metastases. PMID- 9741330 TI - Bone loss induced by cancer treatment and its management. PMID- 9741331 TI - Bisphosphonates. PMID- 9741332 TI - Modulation of virus-induced delayed-type hypersensitivity by plasmid DNA encoding the cytokine interleukin-10. AB - This report evaluates the efficacy of eukaryotic expression plasmids encoding cytokines at modulating the induction and expression of cutaneous delayed-type hypersensitivity (DTH) responses to virus infections. Mice given a single intramuscular administration of cytokine DNA were subsequently infected with either herpes simplex virus (HSV) or vaccinia virus, then tested for DTH. Responses in animals given interleukin-10 DNA were markedly suppressed for at least 5 weeks after pretreatment. Animals also expressed diminished T-cell proliferative responses and modest changes in the balance of T helper type 1 and 2 T-cell reactions. Treatment of animals already sensitized to express DTH, also showed inhibited responses, these taking 6-7 days after treatment to become apparent. Our results show the potency and convenience of plasmid DNA encoding cytokines to modulate inflammatory reactions. Advantages and risks of the cytokine DNA approach are briefly discussed. PMID- 9741333 TI - Potentiation of antigen-specific, Th1 immune responses by multiple DNA vaccination with an ovalbumin/interferon-gamma hybrid construct. AB - The preferential differentiation of T helper (Th) cells to Th1 or Th2 subsets is important with respect to susceptibility or resistance to particular infections, or to autoimmune diseases and allergic diseases. To more effectively drive immune responses toward antigen-specific Th1 responses, we constructed a mammalian expression vector (pOVA/IFN-gamma) carrying a hybrid gene in which the ovalbumin (OVA) (a model antigen) cDNA was covalently linked to murine interferon-gamma (IFN-gamma) cDNA. Intramuscular injection of BALB/c mice with the pOVA/IFN-gamma DNA increased both the production of OVA-specific IFN-gamma by CD4+ T cells and the ratio of anti-OVA immunoglobulin G (IgG) 2a to IgG1 isotypes, while the injection with the pOVA alone, or with the mixture of the pOVA and pIFN-gamma, caused no or little increase. Furthermore, the OVA-specific, Th1 immune responses were dramatically augmented by multiple injections with the pOVA/IFN-gamma DNA. These studies indicate that the direct linkage of an OVA gene to an IFN-gamma gene in the expression plasmid is required for efficiently confining the Th1 effects of IFN-gamma to the OVA-specific cells, and the linkage effect of the OVA/IFN-gamma DNA can be potentiated by multiple vaccination. PMID- 9741334 TI - TCR-alpha chain-like molecule is involved in the mechanism of antigen-non specific suppression of a ubiquitin-like protein. AB - Although existence of suppressor T cells is a controversial issue in cellular immunology, several lines of evidence indicate that T-cell-receptor alpha-chain (TCR-alpha) is a critical component of suppressor factors produced by these cells. Monoclonal non-specific suppressor factor (MNSF), a lymphokine produced by murine T-cell hybridoma, possesses pleiotrophic antigen-non-specific suppressive functions. Recently, we have shown that the 70,000-MW MNSF comprises an 8000-MW ubiquitin-like polypeptide and other subunit(s). Here we report that the 8000-MW ubiquitin homologue is associated with an intracellular TCR-alpha (but not TCR beta)-like molecule and released from the cells. The affinity eluates obtained from the culture supernatants of E17 cells and concanavalin A (Con A)-activated splenocytes with anti-TCR-alpha monoclonal antibody (mAb) showed an antigen-non specific, major histocompatibility complex (MHC)-non-restricted suppression. Immunoblot analysis demonstrated that anti-TCR-alpha, but not anti-TCR-beta, mAb recognizes native 70,000-MW MNSF. In addition, we found the dissociation of the 8000-MW polypeptide from the 62,000-MW TCR-alpha cross-reactive protein by hydrolase which cleaves isopeptide bonds. Thus the covalent attachment of ubiquitin-like protein(s) may be involved in the underlying mechanism of suppressor T-cells and TCR-alpha-like molecule(s) might be a main link between antigen-specific and non-specific suppression. PMID- 9741336 TI - Molecular characterization of the putative T-cell receptor cavity of the superantigen staphylococcal enterotoxin B. AB - A number of investigators have utilized a variety of methods to identify the structural basis for the interaction of superantigens with the T-cell receptor beta-chain. The previous studies strongly suggest that a region of the toxin near residues N23, Y61, Y91 and D209 is important for this binding activity. Examination of crystal structure data shows that these residues line the rim of one side of a shallow cavity in the toxin. In an attempt further to define the face of the staphylococcal enterotoxin B (SEB) molecule involved in the interaction with the beta-chain, we have employed a polymerase chain reaction (PCR)-based, site-specific mutagenesis method to generate amino acid substitutions of residues on the opposite side of this putative T-cell receptor interaction cavity. Our results show that Y175 and N179 appear to be involved in the function of this superantigen, since each of several substitutions at this position exhibits a significantly reduced ability to induce T-cell proliferation. At the same time, mutation of the proximal Y186 does not alter the superantigen activity of SEB. Binding analysis of these mutants shows that class II binding activity is not significantly altered. Analysis of the responding T cells shows that the mutant toxins maintain T-cell receptor V beta selectivity. However, responses of T cells bearing the V beta 8.1 allele appear to be particularly diminished. When viewed in the context of other results reported in the literature, our results suggest that the T-cell receptor interaction site involves SEB residues which ring both the Y175/N179-side and the N23-side of a cavity on one side of the toxin molecule. PMID- 9741335 TI - An allogeneic microenvironment influences the phenotype of intermediate T-cell receptor cells expanding in MRL-lpr/lpr mice. AB - MRL-lpr/lpr (lpr) mice fall victim to autoimmune disease owing to a lymphoproliferative disorder mainly of double-negative (DN) CD4- CD8- alpha beta T cells expressing a low density of interleukin-2 receptor beta-chain (IL-2R beta). It was previously revealed that the lpr gene is a defective Fas gene, into which an early transposon (ETn) of retrovirus is transfected. As a result of the failure of apoptosis, intermediate T-cell receptor (TCR) cells (i.e. TCRint cells) with DN phenotype abnormally accumulate in the periphery of lpr mice. We investigated herein how these TCRint cells are selected in terms of CD4, CD8 and TCR in lpr mice. When a whole fraction of mononuclear cells (MNC) in various immune organs of lpr mice was injected into scid mice (allogeneic circumstance), CD8+ TCRint cells mainly expanded. They had a high density of IL-2R beta. This was true when bone marrow cells of lpr mice were injected into scid mice. On the other hand, when MNC of the spleen and bone marrow in lpr mice were injected into irradiated (9 Gy) lpr mice (syngeneic circumstance), the major expanding cells were DN TCRint cells expressing a low density of IL-2R beta. A cell-sorting experiment for purified fractions demonstrated that only CD8- cells reconstituted TCRint cells in scid mice. Namely, DN CD4- CD8- cells as well as CD4+ cells which once acquired the mature phenotype, no longer switched their phenotype. These results suggest that the phenotype of TCRint cells is influenced by the surrounding microenvironment. PMID- 9741337 TI - Low CD3+CD28-induced interleukin-2 production correlates with decreased reactive oxygen intermediate formation in neonatal T cells. AB - The capacity of neonatal T cells to secrete interleukin-2 (IL-2) has been reported to be variable. We analysed IL-2 production in purified neonatal and adult T cells using polyclonal activator phorbol ester + calcium ionophore (PDBu + iono) or receptor-mediated anti-CD3/anti-CD3+ anti-CD28 stimulation. PDBu + iono induced equally high IL-2 levels in both groups and, when stimulated with plate-bound anti-CD3 monoclonal antibody (mAb), the IL-2 secretion by neonatal cells was undetectable and adult cells produced low amounts of IL-2 (mean 331 +/- 86 pg/ml). The addition of anti-CD28 mAb to anti-CD3-stimulated cells markedly increased IL-2 production in both cell types, but levels of IL-2 in neonatal T cells remained clearly lower than those of adult T cells (respective mean values: 385 +/- 109 pg/ml and 4494 +/- 1199 pg/ml). As NF-kappa B is a critical transcription factor in the control of IL-2 expression, we next analysed its nuclear translocation in neonatal and adult T cells using the electrophoretic mobility shift assay and, because induction of reactive oxygen intermediates (ROI) is required for the activation of NF-kappa B, we also analysed levels of intracellular ROI in these cells using the ROI-reactive fluorochrome DCFH-DA and flow cytometry. In neonatal T cells NF-kappa B activation and ROI formation after anti-CD3 stimulation were low compared with adult T cells and, although addition of anti-CD28 mAb increased induction of NF-kappa B and ROI formation, levels similar to those of adults were not achieved. After PDBu + iono stimulation, the cells showed similar ROI formation and IL-2 secretion. Our results suggest that reduced IL-2 production by neonatal T cells is specific for anti-CD3 and anti CD3+ anti-CD28-mediated stimulation and that these activators cannot effectively activate the ROI-NF-kappa B signalling pathway in neonatal T cells. PMID- 9741338 TI - Activation-dependent expression of the insulin-like growth factor binding protein 2 in human lymphocytes. AB - The expression of the insulin-like growth factor binding protein-2 (IGFBP-2) was assayed in mononuclear cells originating from different organs of the immune system. All mononuclear cells studied did express IGFBP-2, but the expression level was found to be dependent on the cell type and origin of the cell. T cells showed a higher expression of IGFBP-2 mRNA than did B cells, and CD34+ stem cells expressed IGFBP-2 mRNA at a high level. Expression was highest in bone marrow and thymus. Stimulation of peripheral mononuclear cells resulted in a marked increase of IGFBP-2 mRNA and also intracellular IGFBP-2, as analysed by fluorescence staining. This increase parallels the increase of other known T-cell activation markers. Furthermore, the increase of intracellular IGFBP-2 seems to precede T cell blast formation and all T cells in active phases of the cell cycle have high levels of IGFBP-2. Our results provide a basis for further investigations on the contribution of the IGF-system to the regulation of T-cell proliferation and differentiation. IGFBP-2, in particular, may have an important influence in the regulation of T-cell activation and proliferation. PMID- 9741339 TI - Intestinal IgA plasma cells of the B1 lineage are IL-5 dependent. AB - Two lineages of B cells, designated B1 and B2 cells, have been identified based upon their origins, anatomical distribution, cell surface markers, antibody repertoire and self-replenishing potential. B1 cells are maintained by self renewal of cells resident in the peritoneal cavity (PerC) and they utilize a limited repertoire of germline V-region genes, mostly directed against ubiquitous bacterial antigens such as phosphoryl choline (PC). B2 cells are replenished from bone marrow precursors and use a larger repertoire of immunoglobulin V-region genes. Whereas most immunoglobulin A (IgA) plasma cells in the intestine derive from B2 lineage precursors in the Peyer's patch, a subpopulation of Per C-derived B1 cells populate the intestinal lamina propria where they mature into IgA plasma cells. In previous in vivo studies we have shown that whereas IgA+ B2 cells are interleukin (IL)-6 dependent, B1 cells are IL-6 independent. In view of the in vitro evidence that IL-5 is also involved in IgA expression, in the studies reported here we have used IL-5-deficient mice to evaluate the role of IL-5 in vivo in IgA expression in the gut. The results demonstrate that although total IgA cell numbers are only marginally depressed in IL-5-deficient mice, there is a marked selective depletion of IgA+ cells of the B1 lineage in the gut and a corresponding depression in the capacity of these mice to mount an intestinal response to a B1 antigen (PC) but not to a B2 antigen (oralbumin; OVA), reflecting intact B2-derived IgA cell function but a defect in the B1 cell contribution to IgA responses in IL-5 deficient mice. Collectively these data demonstrate differential cytokine regulation of subsets of IgA+ cells in the gut in that IgA+ cells of the B2 lineage are IL-6 dependent but IL-5 independent, but B1-derived IgA+ cells are IL-5 dependent and IL-6 independent. PMID- 9741340 TI - A 24,000 MW Trypanosoma cruzi antigen is a B-cell activator. AB - Trypanosoma cruzi, the causative agent of Chagas' disease, is a protozoan parasite that infects humans and other mammals in Central and Latin America. Several alterations of the immune response after infection have been described, such as severe immunosuppression of both cellular and humoral responses and massive polyclonal B- and T-cell activation, including the expansion of self reactive clones. We have investigated the effects of the intraperitoneal injection of a recombinant 24,000 MW T. cruzi-specific antigen (rTc24) on the immune response of normal and deficient strains of mice. We analysed the in vivo and ex vivo levels of lymphocyte activation and the proliferative responses to rTc24 by determining the expression of CD69 activation marker and the levels of thymidine incorporation by spleen cells. The numbers of antibody-producing cells were determined by ELISPOT and the levels of immunoglobulin in the sera by isotype-specific enzyme-linked immunosorbent assay. We observed an increased [3H]thymidine ([3H]TdR) incorporation by spleen cells after rTc24 stimulation in vivo and in vitro. This proliferative activity induced by rTc24 was independent of the mouse strain used in the experiments (including C3H/HeJ mice) and ruled out the possibility that rTc24 preparations were contaminated by lipopolysaccharide. The injection of rTc24 protein induced preferentially the activation of B cells, as determined by the increased expression of CD69 molecules on IgM+ spleen cells. Considerable increases of IgM-secreting B cells were determined in both athymic and euthymic BALB/c mice. Mice that are deficient in B cells (BALB.Xid) responded to rTc24 but to a lesser extent. These increases in IgM B-cell numbers were accompanied by elevated levels of IgM immunoglobulins in the sera of injected animals. Our results suggest a role for rTc24 in B-cell activation. PMID- 9741341 TI - Changes in cellular response to mycobacterial antigens and cytokine production patterns in leprosy patients during multiple drug therapy. AB - Changes in Mycobacterium leprae-induced lymphoproliferative responses and mediator release by leprosy patients' lymphocytes were followed during multiple drug therapy (MDT). At the time of diagnosis, multibacillary (MB) patients who did not develop reactions responded to both sonicated M. leprae and synthetic disaccharide coupled to bovine serum albumin (ND-BSA) antigens, but those who would later develop reactions did not respond, even in the presence of added cytokines. The paucibacillary (PB) group initially had high responses to sonicated M. leprae but no response to ND-BSA, even in the presence of added cytokines. In the first year of treatment, the supernatants of PB patients' cell cultures contained factors that enhanced the phytohaemagglutinin (PHA) response of normal cells. In contrast, those MB patients who did not develop reactions at a later stage produced culture supernatants that were inhibitory. Interestingly, the MB patients who later developed reactions during treatment, and did not initially respond to M. leprae, produced supernatants containing enhancing factors, like those of the PB group. Later on in the treatment, all patients had the same patterns: when response to M. leprae decreased from its highest level, inhibitory factors were produced. Further studies revealed that the supernatants which inhibited the PHA response of normal cells contained the active form of transforming growth factor-beta 1, (TGF-beta 1), whatever the disease type or treatment status of the donor. These TGF-beta 1 levels correlated directly with the degree of inhibition. Similarly supernatants that neither inhibited nor enhanced PHA responses contained the highest levels of interleukin-10 (IL-10), while those from treated patients that enhanced contained the lowest levels of interleukin-4 (IL-4) and interferon-gamma (IFN-gamma). These cytokine correlations transcended the conventional disease classification, and imply that all patients pass through a sequence of patterns of immune response during treatment. These treatment-induced changes may explain occasional reports of response patterns at variance with the 'immunological spectrum' of leprosy. PMID- 9741342 TI - Characterization of a monoclonal antibody that recognizes a lymphocyte surface antigen for the cetacean homologue to CD45R. AB - As part of our current efforts to develop assays and reagents to study the immune system of marine mammals, and in view of the effort currently made to develop monoclonal antibodies to cell surface proteins of lymphocyte subsets in different species, the present paper reports on the characterization of a monoclonal antibody against the homologue of CD45R on cetacean lymphocytes. The specificity of this antibody has been characterized on the basis of immunoprecipitation of the antigen it recognized, immunoperoxidase staining on cetacean lymph node and thymus sections, as well as one and two-colour flow cytometric analysis of cetacean peripheral blood mononuclear cells and single-cell suspensions of thymus, lymph node and spleen. Anticetacean CD45R (F21.H) immunoprecipitated proteins of 180, 200 and 220 x 10(3) MW, with the 180 x 10(3) MW from being predominantly expressed on T cells and the 220 x 10(3) MW form expressed predominantly on B cells and thymocytes F21.H labelled all B cells and a proportion of T cells on single-cell suspensions of spleen cells. CD45R- killer whale peripheral blood lymphocytes expressed a higher density of CD2 than CD45R+, a characteristic of memory T cells. Killer whale T lymphocytes also lost the expression of CD45R upon activation with concanavalin A (Con A) and phytohaemagglutinin (PHA). This is the first report of a monoclonal antibody to CD45R in cetaceans, and this antibody is foreseen as a possible valuable diagnostic and research tool to assess immune functions of captive and wild cetaceans as part of the evaluation of their health status. PMID- 9741343 TI - Novel mode of action of the calcium antagonist mibefradil (Ro 40-5967): potent immunosuppression by inhibition of T-cell infiltration through allogeneic endothelium. AB - Cyclosporin A reduces the mitotic activity of allosensitized lymphocytes, but fails to limit emigration of these cells into the donor organ. However, the modulation of both lymphocyte proliferation and infiltration are desirable characteristics of immunosuppressive therapy. The calcium-channel blocker, verapamil, has recently been shown to effectively prevent the transmigration of CD4+ and CD8+ T cells through allogeneic endothelium. Mibefradil (Ro 40-5967) represents a new generation of calcium antagonists with high potency and long term activity. To evaluate the immunosuppressive potential of this drug, the influence of mibefradil on lymphocyte adhesion to, horizontal locomotion along, and penetration through allogeneic endothelium (HUVEC) was performed. When lymphocytes were prestimulated for 24 hr with mibefradil, adhesion and penetration were dose-dependently reduced. The adhesion ID50 values were 3.4 microM (CD4+ T cells) versus 9.2 microM (CD8+ T cells) and 2.1 microM (CD4+ T cells) versus 3.9 microM (CD8+ T cells) with regard to penetration. Mibefradil also effectively blocked horizontal locomotion. Specific down-regulation of T cell binding to the P-selection receptor (ID50: CD4+ T cells, 0.8 microM: CD8+ T cells, 1.2 microM) and to the intracellular adhesion molecule-1 (ICAM-1) receptor (ID50: CD4+ T cells, 1.9 microM; CD8+ T cells, 1.5 microM) by mibefradil seems to be responsible for the decreased adhesion and penetration rates. Reduction of intracellular F-actin in T lymphocytes could diminish cell locomotion. In conclusion, the potent suppressive properties of mibefradil support its use as a co-medication in cyclosporin A-based immunosuppressive therapy. PMID- 9741344 TI - The effects of monocytes on the transendothelial migration of T lymphocytes. AB - In vivo cell-mediated immune reactions are characterized by mixtures of monocytes and T cells. The purpose of this study was to investigate the role of monocytes on T-cell migration and induction of endothelial adhesion molecules. The in vitro model consisted of adding peripheral blood mononuclear cells (PBMC), T cells or mixtures of monocytes and T cells, to endothelial cells on a porous membrane and using flow cytometry to distinguish between the monocyte and lymphocyte components. PBMC and PBMC supernatants were highly potent at upregulating intercellular adhesion molecule-1 (ICAM-1) and inducing expression of vascular cell adhesion molecule-1 (VCAM-1) and E-selectin. Induction by supernatants was inhibited by antibodies to tumour necrosis factor-alpha (TNF-alpha) and interleukin-1 (IL)-1 beta. Using monocyte-enriched populations, as few as one monocyte to 100 endothelial cells was sufficient to upregulate adhesion molecules. Fixed monocytes also induced adhesion molecules and expressed surface bound cytokines. In contrast, highly purified unstimulated T cells were not found to induce adhesion molecules at 4, 6, 24 or 48 hr of coculture. Purified T cells showed low-level migration through resting (VCAM-1 negative) endothelium, which was approximately doubled by addition of small numbers of monocytes or TNF-alpha. In conclusion, monocytes, via cell surface or released cytokines play an essential role in allowing large-scale recruitment of T cells to inflammatory sites in vivo. PMID- 9741345 TI - Prostaglandin and fatty acid modulation of Escherichia coli O157 phagocytosis by human monocytic cells. AB - Phagocytosis by human monocytes is an important primary survival mechanism particularly during bacterial infection. However, the processes that control the events and mediators involved in the activation of monocytes and their impact on the phagocytosis of bacteria are poorly understood. The effect of bacterial endotoxin, interleukin-1 beta (IL-1 beta), fatty acids and prostaglandin E2 (PGE2) on the phagocytosis of fluoroscein isothiocyanate (FITC)-labelled Escherichia coli (O157) by human blood monocytes and U937 cells was studied by flow cytometry. Endotoxin increased the phagocytosis of labelled bacteria by both monocytes and U937 cells. IL-1 beta and the polyunsaturated fatty acids; dihomo gamma-linolenic and arachidonic acids also increased the phagocytic activity of both monocytes and U937 cells. In contrast, PGE2 suppressed phagocytosis in a concentration-dependent manner. The cyclo-oxygenase inhibitor, ketoprofen, further enhanced the increased phagocytic activity in the presence of endotoxin and interleukin-1 (IL-1) indicating suppression by endogenous prostaglandins. This was confirmed by the data which showed that lipopolysaccharide (LPS) and IL 1 increased PGE2 release and ketoprofen inhibited release. Endotoxin and fatty acids increased IL-1 beta release also, whereas PGE2 inhibited release. The data suggest that phagocytic activity may be linked to changes in IL-1 levels. The data presented in this study also suggest that monocyte phagocytosis in the course of bacterial infection would be altered during pathophysiological events which result in elevation of extracellular fatty acids. PMID- 9741346 TI - Expression of the TrkB neurotrophin receptor by thymic macrophages. AB - Increasing evidence suggests that some members of the neurotrophic factor family of neurotrophins could be implicated in the regulation of immune responses. Neurotrophins, as well as their tyrosine kinase signal-transducing receptors (the so-called Trk neurotrophin receptors), have been detected in different lymphoid tissues, although their cellular localization is not well known. In this study we used single and double immunohistochemistry to localize TrkB in situ in the rat thymus (in animals from 0 days to 2 years of age), in cytospin preparations of rat thymic cells, and in two mouse monocyte-macrophage cell lines (RAW 264.7 and J774A.1). We found TrkB protein expression in a subpopulation of cells in the corticomedullary junction, which simultaneously expressed the rat macrophage marker ED1. The density of TrkB-expressing cells increased with age, reaching maximal values at 2 years. Conversely, no evidence of TrkB protein expression could be found in dendritic cells, epithelial cells or thymocytes. Thymic macrophages in cytospin preparations, as well as in the mouse monocyte macrophage cell lines, also expressed TrkB protein. Although the possible function of TrkB in the thymic macrophage remains to be clarified, present findings add further evidence to the proposed role of neurotrophins in the immune system. PMID- 9741347 TI - Morphological alterations in rat peritoneal mast cells by stem cell factor. AB - Stem cell factor (SCF) stimulates mast cell adhesion and, because SCF is produced normally in tissues, it may be a major factor responsible for the adhesion of mast cells to connective tissue matrix. We found that the morphology of rat peritoneal mast cells (RPMC) altered after the addition of recombinant murine SCF (rmSCF) in vitro. The ability of rmSCF to enhance morphological alteration was dose dependent and completely abolished by anti-c-kit ACK2 monoclonal antibody. Exposure of RPMC to transforming growth factor-beta 1, wortmannin, genistein, herbimycin A, staurosporine, indomethacin and cytochalasin D before the addition of rmSCF antagonized rmSCF-induced morphological alteration. However, nordihydroguiaretic acid had no effect. Many RPMC appeared to respond also to nerve growth factor (NGF) but the total number of cells with altered morphology was much greater when the culture was stimulated by rmSCF than by NGF. We suggest that morphological alterations of mast cells by rmSCF is an important step for the participation in adhesion to tissue under resident physiological conditions. PMID- 9741348 TI - Staphylococcal enterotoxin B inhibits the production of interleukin-4 in a human mast-cell line HMC-1. AB - Staphylococcal enterotoxins belong to the recently characterized group of immunocytotropic bacterial superantigens that are potent mitogens for human T cells. Superantigens are presented, but without intracellular processing, to T cells by monocyte/macrophages, Langerhans' cells and keratinocytes via the class II major histocompatibility complex (MHC) molecules. Superantigens have been demonstrated to act as potent inducers of several proinflammatory cytokines in the antigen-presenting cells such as interleukin-1 (IL-1) and tumour necrosis factor-alpha (TNF-alpha). As mast cells participate in the pathogenesis of several inflammatory skin disorders such as atopic dermatitis (AD), which is often aggravated by staphylococcal infections, we studied the effect of staphylococcal enterotoxin B (SEB) superantigen on the histamine release and IL-4 expression in a human mast-cell line (HMC-1). Incubation of SEB (50 micrograms/ml) with HMC-1 cells for 45 min, could not induce any histamine release. The HMC-1 cells were also stimulated with various concentrations of SEB (0, 1, 10, 20, 50 micrograms/ml) for 1, 2, 3 and 4 days. Clear dose-dependent inhibition of IL-4 protein production and release was observed on day 4 without any observed effect on cell viability. Compared with unstimulated HMC-1 cells, after 50 micrograms/ml SEB stimulation, the IL-4 mRNA levels decreased steadily in the 2, 6, 18 and 24 hr samples in repeated experiments as measured with the reverse transcription polymerase chain reaction (RT-PCR) method. In comparison, a biphasic decrease in TNF-alpha expression was found. Our results show that an human leukaemic mast cells, superantigen stimulation downregulates the production of IL-4. PMID- 9741349 TI - Biological activities of C3 beta c, a novel neutrophil chemoattractant derived from the beta-chain of rat complement C3. AB - Biological activities of C3 beta c, which is a C-terminal fragment of the beta chain of rat complement C3, have been studied by in vivo and in vitro experiments. C3 beta c was purified as a novel neutrophil chemoattractant from the exudate of the chronic phase of rat carrageenin-induced inflammation. The purified C3 beta c induced neutrophil chemotaxis in vivo when C3 beta c was injected into the preformed air-pouch on the back of rats. C3 beta c transiently increased the intracellular free Ca2+ concentration of neutrophils and enhanced the adhesion of neutrophils to fibrinogen in vitro, suggesting that C3 beta c has the ability to express an adhesion molecule of rat neutrophils. In addition, C3 beta c at low concentrations (10(-10)-10(-11) M) stimulated rat macrophages to produce cytokine-induced neutrophil chemoattractant-2, a member of the interleukin-8 family. Furthermore, C3 beta c enhanced vascular permeability in vivo, which is suppressed by cyproheptadine, suggesting that C3 beta c may have the characteristics of an anaphylatoxin. Our results suggest that C3 beta c contributes to oedema formation and neutrophil accumulation at inflammatory sites in rats. PMID- 9741350 TI - Soluble Fas mRNA is dominantly expressed in cases with silicosis. AB - Although it is well known that cases with silicosis exhibit various immunological abnormalities, the mechanisms involved in the occurrence of immuno-dysfunction or dysregulation induced by silica compounds have not yet been determined. Fas is a well-known cell surface molecule that is involved in the apoptosis pathway that belongs to the tumour necrosis factor-receptor family. Soluble Fas (sFas) is produced as an alternatively spliced product of the Fas gene and protects cells from apoptosis due to antagonization of the binding between membrane form of the Fas gene (mFas) and the Fas ligand. To determine the role of the Fas/Fas ligand system in silica-induced immunological abnormalities, we investigated Fas and Fas ligand message expression levels using the multiplex reverse transcription polymerase chain reaction (RT-PCR) method with peripheral blood mononuclear cells from silicosis cases with no clinical symptoms of autoimmune diseases. Although the relative expression levels of the Fas or Fas-ligand genes were not remarkably altered in these cases, we observed the sFas message was dominantly expressed compared with mFas expression. These results suggest that self-recognizing clones in cases with silicosis survive for decades, escaping the exclusion mechanisms induced by apoptosis. Then they cause the appearance of autoantibodies and the acquisition of autoimmune diseases sequentially. PMID- 9741351 TI - Wild isolates of murine cytomegalovirus induce myocarditis and antibodies that cross-react with virus and cardiac myosin. AB - The laboratory-adapted K181 strain of murine cytomegalovirus (MCMV) induces both acute and chronic myocarditis, associated with autoantibodies to cardiac myosin, in susceptible BALB/c mice. However, the K181 MCMV strain has been maintained in the laboratory for many years and may not resemble naturally occurring strains of MCMV in its ability to induce myocarditis. Accordingly, six different isolates of MCMV from wild Mus domesticus were compared with K181 MCMV for their ability to induce myocarditis and autoantibodies to cardiac myosin in BALB/c mice. These isolates were shown to induce acute myocarditis similar to K181 MCMV, with associated focal and diffuse myocardial inflammation. However, the levels of myocarditis induced by the wild isolates during the chronic phase of the disease (days 32-56 post-infection) were low in contrast to the K181 strain. Interestingly, 30% of wild-trapped mice showed histological evidence of myocarditis and all were sero-positive to MCMV. Sera from BALB/c mice infected with wild MCMV isolates and from wild-trapped mice contained antibodies that cross-reacted with MCMV and cardiac myosin (S2 region). The cross-reactive region of MCMV was found to be a 50,000-55,000 MW viral polypeptide. These findings suggest that molecular mimicry may be involved in the pathogenesis of autoimmune myocarditis following infection with both laboratory and wild MCMV strains. PMID- 9741352 TI - Uveitogenic epitopes of retinal S-antigen are generated in vivo via an alternative antigen-presentation pathway. AB - We have found that different antigen-processing pathways are involved in the induction of experimental autoimmune uveoretinitis (EAU) by the retinal autoantigens S-antigen and interphotoreceptor retinoid-binding protein (IRBP). Although in vitro T-cell proliferative responses to IRBP were completely inhibited in the presence of irreversible cysteine protease inhibitors, no significant reduction of S-antigen proliferative responses was found. Furthermore, acidic proteolysis of S-antigen by the cysteine protease cathepsin B prior to immunization radically reduced pathogenicity (disease severity). In addition, in vitro processing of S-antigen, but not IRBP, was also found to be resistant to the action of cycloheximide and lysosomotropic agents, inhibition of proliferation only occurring after extended exposure of antigen-presenting cells to methyl amine or high concentrations of chloroquine. These data indicate that an alternative pathway of antigen processing exists for S-antigen, which is independent of processing within the normal endolysosomal pathway and that uveitogenic peptides of naturally processed S-antigen bind to major histocompatibility complex class II antigens either at the cell surface or within very early endosomes where cathepsin B is inactive. PMID- 9741353 TI - Modulation of alloimmune response in vitro by an IgM-enriched immunoglobulin preparation (Pentaglobin). AB - The mode of action of intravenous immunoglobulins (IVIG) in autoimmune and immunoregulatory disorders is still poorly understood. In vitro, direct effects of IVIG on cytokine release and on cytokine receptors have been described, as well as naturally occurring, neutralizing anticytokine antibodies. The aim of our study was to investigate whether the enrichment in IgM and IgA would have any impact on the in vitro immunomodilatory capacity of IVIG. The preparation tested (Pentaglobin) contains 76% IgG and 12% IgM and IgA, respectively. We could demonstrate a significant inhibition of alloantigen-induced proliferation in the mixed lymphocyte reaction (MLR) even at a Pentaglobin concentration of 1.0 mg/ml. About 10-fold higher concentrations of standard 7S IVIG containing only trace amounts of IgM and IgA were necessary to achieve equivalent suppression of the alloimmune response. Similarly, phytohaemagglutinin (PHA)-induced lymphocyte proliferation was more effectively inhibited by Pentaglobin than by standard 7S IVIG. Cytokine analyses in culture supernatants of MLR provide evidence that Pentaglobin not only modulates interleukin-2 (IL-2), which has already been observed with standard 7S IVIG, but, moreover, modulates interferon-gamma production with a subsequent impact on monocyte-derived tumour necrosis factor alpha and IL-6 release. Based on these results we conclude that in vitro the IgM- and IgA-enriched Pentaglobin has a more potent immunomodulatory capacity than conventionally used standard 7S IVIG. PMID- 9741354 TI - Expression of the heat shock protein 47 in gentamicin-treated rat kidneys. AB - Heat-shock proteins (HSPs) are rapidly synthesized in cells in response to various cytotoxic agents. Although several stress proteins are actively involved in the gentamicin-induced renal damages, the possible role of HSP47 in this condition is not yet clear. In this study, the expression of HSP47 in the gentamicin nephrotoxicity was examined by immunohistochemistry. Twenty male Wistar rats were sacrificed at day 0, 3, 7, 14 and 28 after subcutaneous injection of gentamicin. Gentamicin treatment causes tubular necrosis at day 3, followed by tubular regenerative changes and interstitial fibrosis, which was most prominent at day 14. The renal structures returned to almost normal architectures at day 28. By immunohistochemistry, HSP47 was weakly expressed in most of the glomeruli and occasionally in interstitial cells in the control rat kidneys. In contrast, strong immunostaining for HSP47 was noted in the tubular epithelial cells and interstitial cells in gentamicin treated rat kidneys, and strongest staining was observed at day 7. The immunostaining for HSP47 then gradually decreased, and returned to the normal level at day 28. In the whole experimental period staining pattern of HSP47 in the glomeruli was not changed. In addition, phenotypically altered tubulointerstitial cells including regenerative tubular epithelial cells (immuno-positive for vimentin) and interstitial cells (immuno-positive for alpha-smooth muscle actin) were found in gentamicin nephrotoxicity. Expression of type III collagen increased in the areas of interstitial fibrosis. By double immunostaining, the regenerated and phenotypically altered tubulointerstitial cells were found to express HSP47 in and around interstitial fibrosis. It is concluded that overexpression of HSP47 by phenotypically altered renal cells might play a significant role in the development of gentamicin nephrotoxicity. PMID- 9741355 TI - Expression of heat shock protein 47 is increased in remnant kidney and correlates with disease progression. AB - Glomerulosclerosis is characterized by accumulation of the mesangial extracellular matrix, including type I and V collagen. The processing for the collagens in the glomeruli may play a critical role for development of glomerulosclerosis. We examined the expression of heat shock protein 47 (HSP47), a collagen-binding molecular chaperone in the progressive glomerulosclerosis model. Subtotally nephrectomized rats, unlike sham-operated rats, developed focal and segmental glomerulosclerosis. Immunological staining demonstrated an increased expression of HSP47 which paralleled the expression of type I and IV collagen in the glomeruli of the nephrectomized rats as the glomerulosclerosis developed. The mRNA levels encoding type I and type IV collagen and HSP47 were increased 3.4 fold, 3.6 fold and 2.8 fold, respectively, at week 7 after nephrectomy. By in situ hybridization, the expression of HSP47 mRNA was determined to be localized to the glomeruli with segmental sclerosis. These results suggest that HSP47 may play a central role in the process of extracellular matrix accumulation during the development of glomerulosclerosis. PMID- 9741356 TI - Glycated plasma proteins in experimentally induced acute toxic renal failure by dichromate injection: evidence for loss with urine and decreased plasma levels. AB - In the rat, a single subcutaneous injection of sodium dichromate (20 mg/kg) causes acute renal injury and significant polyuria, proteinuria, and glycosuria (peaking 2-3 days after treatment, and returning to normal by day 5) without any changes in the plasma levels of protein, glucose, and glycated haemoglobin. Surprisingly, the percentage levels of glycated plasma total proteins and albumin (assayed by boronate affinity chromatography) transiently and significantly decrease during recovery from proteinuria (days 4 and 10 after treatment) and were found in the normal range of values by day 18. These changes are concomitant with a significant increase in the percentage level of glycated albumin in urine. Constancy of total plasma protein and the temporal pattern of levels of glycation suggest that changes in the percentage values of glycated proteins are secondary to a transient selective loss of glycated plasma proteins in urine. PMID- 9741357 TI - Gadolinium chloride removes pulmonary intravascular macrophages and curtails the degree of ovine lentivirus-induced lymphoid interstitial pneumonia. AB - Ovine lentivirus (OvLV), a retrovirus, causes lymphoid interstitial pneumonia (LIP) in sheep. Pulmonary alveolar macrophages are believed to play a central role in lung inflammation caused by this virus. The pulmonary intravascular macrophage (PIM), a recently identified inflammatory cell, is under active investigation for its role in lung pathology. Gadolinium chloride (GC), a rare earth lanthanide, has been used in in vivo studies to abrogate macrophage function to understand their role in tissue pathology. We treated mock- and OvLV infected lambs with GC once a week for 34 weeks to investigate the implications of PIMs in the pathogenesis of LIP using light and electron microscopy. Repeated treatments with GC did not cause any apparent physiological abnormalities in the lambs. Light microscopy on tissue sections showed that GC reduced the number of PIMs in OvLV-infected and mock-infected lambs. Ultrastructural examination of lung tissues from GC-treated lambs revealed electron-dense deposits of GC-like material in endosomes and cytoplasm of PIMs. The majority of the PIMs in GC treated lambs appeared damaged. Semi-quantitative histological evaluation showed amelioration of the extent of LIP in GC-treated OvLV-infected lambs compared to placebo-treated OvLV-infected lambs. This study stresses the effectiveness of GC in eliminating PIMs and suggests their involvement in the pathogenesis of OvLV induced LIP. The results also strengthen the usefulness of GC as a tool to study in vivo function of cells such as PIMs that are difficult to isolate for in vitro studies. PMID- 9741358 TI - Regional activation of the immediate-early response gene c-fos in infarcted rat hearts. AB - Regional infarction of the left ventricle is followed by hypertrophy of the viable myocardium. This compensatory growth of cardiac myocytes requires induction of gene transcription and synthesis of proteins. In this study, we examined the expression of the immediate-early response gene c-fos following ligation of the left coronary artery in rat hearts. RNase protection assay demonstrated a rapid increase in the c-fos mRNA level in the ventricular myocardium. After two days of infarction, the c-fos expression was attenuated and was comparable to that observed in sham-operated control hearts. In situ tissue distribution of Fos protein-like immunoreactivity revealed the appearance of positively stained cells adjacent to the lateral border of the ischaemic myocardium, in the left ventricular subendocardial areas, in the papillary muscles of the left ventricle, in the proximity of great transmural vessels, and focally in the normo-perfused subepicardial myocardium. Double staining using antibodies recognizing the Fos protein and alpha-actinin, confirmed that the accumulation of nuclear Fos protein-like immunoreactivity was mainly seen in the cardiac myocytes. However, double staining of the Fos protein and Hoechst DNA labelling showed that detectable immunoreactivity occurred only in a limited proportion of the total nuclei present in these myocardial regions. Moreover, the regions showing c-fos activation correspond to the areas in which the appearance of subsequent growth responses are most pronounced following myocardial infarction. The present results therefore indicate that an early and regional c fos activation is taking place in viable cardiac myocytes following left coronary artery ligation, and that c-fos is a possible regulating factor of sequential events leading to altered pattern of gene expression and protein synthesis in the hypertrophying heart. PMID- 9741359 TI - Vascular deprivation-induced necrosis of the femoral head of the rat. An experimental model of avascular osteonecrosis in the skeletally immature individual or Legg-Perthes disease. AB - The blood supply of rats' femoral heads was severed by cutting the ligamentum teres and stripping the periostium. Histologically, necrosis of the marrow was apparent on the 2nd postoperative day, necrosis of the bone on the 5th postoperative day and fibrous ingrowth on the 7th postoperative day. During the following 5 weeks, progressive resorption of the intertrabecular necrotic debris and necrotic bony trabeculae and subchondral bone plate and, concurrently, appositional and intramembranous new bone formation resulted in remodeling of the femoral heads. In 2 of 7 femoral heads, replacement of the necrotic bone by viable bone was complete at the 42-day postoperative interval. Also, the articular cartilage of the deformed and flattened femoral heads was undergoing degenerative changes. Reduplicating the pathogenically inferred clinical settings of blood supply deprivation, it is proposed that this model, in a small laboratory animal, satisfies the requirements sought for preclinical studies of treatment modalities of avascular osteonecrosis in man. PMID- 9741361 TI - Brittle asthma: fiend or phantom? PMID- 9741360 TI - Comparative histopathology in mouse typhoid among genetically diverse mice. AB - Genetically resistant CBA and A/J mice and susceptible BALB/c and C57BL/6 mice were challenged with either an identical infective dose or a minimal lethal dose of Salmonella typhimurium. The histopathological progression of the disease was examined in tissue sections prepared by the JB-4 Plus resin embedding method and compared between the resistant and susceptible mice. In a fatal disease, the lesions in both animal hosts began with focal abscesses within the first three days post infection. Mononuclear cell infiltration started by day 4 and transformed the lesions into granulomata. Well-formed granulomata were evident by day 7 and persisted in sublethally infected resistant mice. Massive bacterial proliferation and extensive tissue degeneration marked the terminal stage of a lethal challenge. There were no distinguishable features that would identify the tissue response to infection in a resistant host from a susceptible one, except that the lesions in the sublethally infected resistant mice advanced slower and were discrete and self-limiting. PMID- 9741362 TI - Improved survival in ARDS: chance, technology or experience? PMID- 9741363 TI - Lung research funding in the UK: the British Lung Foundation perspective. PMID- 9741364 TI - Rapid onset asthma: a severe but uncommon manifestation. AB - BACKGROUND: Studies of asthma death and severe life threatening asthma (SLTA) include reports of patients who had rapid onset asthma. A study was undertaken to determine the relative frequency of rapid (< 6 hours duration) and slow (> or = 6 hours) onset attacks in patients admitted to hospital with acute severe asthma, and to establish whether those with rapid onset asthma differ in terms of risk factors for asthma morbidity and mortality such as indices of asthma severity/control, socioeconomic factors, health care, and psychological factors. METHODS: A cross sectional study was performed on 316 patients aged 15-49 years admitted with acute severe asthma and interviewed within 24-48 hours of admission. RESULTS: Patients underestimated the duration of the index attack. Only 27 (8.5%) were classified as rapid onset. There were more men in the rapid onset group than in the slow onset group (52% versus 26%), and there was evidence of socioeconomic advantage in the patients with rapid onset attacks. The rapid onset group had more previous episodes of SLTA and were more likely to present with SLTA, but there was no difference in length of stay in hospital. The rapid onset group were less likely to have presented to a GP during the index attack and were more likely to have used ambulance services. There was no difference between the groups in any psychological or health care measure. CONCLUSIONS: Rapid onset attacks are an important but uncommon manifestation of asthma that are more likely to present with SLTA in patients who are more likely to have had previous SLTA. Male subjects are at increased risk of rapid onset attacks, and socioeconomic disadvantage, deficiencies in health care (ongoing and acute), and psychological factors are no more common in these patients than in those with attacks of slow onset. These data are consistent with the hypothesis that there is a small proportion of patients with rapid onset severe asthma who do not have the usual risk factors associated with asthma morbidity or mortality, and thus require different management strategies. PMID- 9741365 TI - Airway wall thickness in patients with near fatal asthma and control groups: assessment with high resolution computed tomographic scanning. AB - BACKGROUND: Airway wall thickening has been observed in post mortem studies of patients with asthma. Assessment of airway wall thickening by high resolution computed tomographic (HRCT) scanning has been reported in experimental studies. We have used HRCT scanning to measure airway wall thickness at the segmental and subsegmental levels in 40 patients with asthma and 14 normal controls. METHODS: The subjects were prospectively divided into four age and sex matched groups: 14 patients with a history of near fatal attack of asthma (NFA; group 1), 12 patients with moderate asthma (group 2), 13 patients with mild asthma (group 3), and 14 normal controls (group 4). All subjects were non-smokers. High resolution (1 mm collimation) CT scans of the chest were done at five different levels. RESULTS: The mean (SD) forced expiratory volume in one second (FEV1) was 68 (20)% of predicted for group 1, 73 (12)% for group 2, 102 (12)% for group 3, and 103 (12)% for group 4. The ratio of airway wall thickness to outer diameter (T/D) and the percentage wall area (WA%) defined as (wall area/total airway area) x 100 were used to compare airway wall thickness between the groups. The mean (SD) T/D and WA% were 0.27 (0.05) and 78.0 (9.2)% for group 1, 0.27 (0.05) and 78.8 (9.2)% for group 2, 0.25 (0.04) and 74.2 (7.5)% for group 3, and 0.23 (0.04) and 70.9 (8.2)% for group 4. T/D and WA% were not significantly different between groups 1 and 2. However, both groups 1 and 2 had higher T/D and WA% than either group 3 or 4 (p < 0.001) and group 3 had a higher T/D and WA% than group 4 (p < 0.03). The differences (95% CI) between the groups in WA% were 7.1% (0 to 14.4) for groups 1 and 4, 3.8% (-3.4 to 10) for groups 1 and 3, and 3.3% (-4.4 to 10) for groups 3 and 4. The differences between the groups in T/D and WA% were noted both for those with airways with a luminal diameter of > 2 mm and those with a luminal diameter of < or = 2 mm. CONCLUSIONS: All the patient groups had greater airway wall thickening than the normal subjects as assessed by HRCT scanning, but patients with more severe asthma had greater airway wall thickening than those with mild asthma. The methodology described in this study may be useful in assessing airway calibre in early intervention studies with anti-inflammatory therapy. PMID- 9741366 TI - Chlamydia pneumoniae and asthma. AB - BACKGROUND: This study was designed to test the association of Chlamydia pneumoniae infection with asthma in a multi-racial population, after adjustments for several potential confounding variables. METHODS: Antibodies to C pneumoniae were measured by microimmunofluorescence in 123 patients with acute asthma, 1518 control subjects admitted to the same hospital with various non-cardiovascular, non-pulmonary disorders, and 46 patients with severe chronic asthma, including some with "brittle" asthma. Acute infection or reinfection was defined by titres of IgG of > or = 512 or IgM > or = 8 or a fourfold rise in IgG, and previous infection by IgG 64-256 or IgA > or = 8. Logistic regression was used to control for likely confounders, including ethnic origin, age, sex, smoking habit, steroid medication, diabetes mellitus and social deprivation, on antibody levels. RESULTS: Antibody titres consistent with acute C pneumoniae infection were found in 5.7% of patients with acute asthma and 5.7% of control patients, while 14.6% of patients with acute asthma and 12.7% of control patients had titres suggesting previous infection. These two groups did not differ significantly. However, titres suggesting previous infection were found in 34.8% of patients with severe chronic asthma: the difference between this group and the control group was statistically significant with an adjusted odds ratio of 3.99 (95% confidence interval 1.60 to 9.97). CONCLUSIONS: These data raise important questions about the previously demonstrated association of C pneumoniae infection with asthma, and suggest that future studies of this association should give particular attention to the presence or absence of a history of severe chronic asthma. PMID- 9741367 TI - Induction of IL-8 production in human alveolar macrophages and human bronchial epithelial cells in vitro by swine dust. AB - BACKGROUND: Exposure to swine dust causes an intense airway inflammation with increased levels of interleukin 8 (IL-8) and predominantly neutrophils in the nasal and bronchoalveolar lavage fluids of healthy human subjects. It is not clear which components in the swine house environment are responsible for the airway reaction. The aim of the present study was to evaluate and compare the effect in vitro of swine dust components on human alveolar macrophages and bronchial epithelial cells. METHODS: Normal human bronchial epithelial cells (NHBE), human pulmonary epithelial carcinoma cell line (A549), and human alveolar macrophages were stimulated with swine dust, lipopolysaccharides (LPS; present in Gram negative bacteria), grain dust (swine feed components), and glucans (a structural component of fungi) in a dose response manner (1-100 micrograms/ml). RESULTS: Swine dust at a concentration of 100 micrograms/ml increased IL-8 production 20 fold in NHBE cells, 28 fold in A549 cells, and 15 fold in macrophages. LPS (100 micrograms/ml) stimulated all three cell types significantly, in macrophages to the same extent as swine dust, but in NHBE and A549 cells swine dust was 5-8 times as potent. Grain dust (100 micrograms/ml) had no effect in A549 cells and macrophages but not NHBE cells. Both glucans and grain dust were weaker stimuli than swine dust and LPS. The LPS content of swine dust solution was 2.16 (0.2) ng/100 micrograms and of grain dust was 0.53 (0.04) ng/100 micrograms. CONCLUSIONS: Swine dust is a strong stimulus for IL-8 production in both bronchial epithelial cells and human alveolar macrophages, whereas LPS has different potency in these cells. PMID- 9741368 TI - Clinical features and prognosis of life time non-smokers with severe alpha 1 antitrypsin deficiency. AB - BACKGROUND: The hereditary disorder alpha 1-antitrypsin deficiency is characterised by development of severe emphysema at an early age with smoking being the most significant additional risk factor. The purpose of the present paper was to analyse potential risk factors other than smoking for emphysema and to estimate the prognosis of life time non-smokers. METHODS: Patients were identified through the files of the Danish alpha 1-antitrypsin deficiency register which contains information on more than 700 persons with the condition. Many of the patients, the non-index cases, were identified from family studies. RESULTS: There were 75 life time non-smokers with PiZ (27 index cases and 48 non index cases) aged 20 years or more at entry. Twenty one subjects died during the follow up period. The Standardised Mortality Ratio (SMR) was 3.0 (95% confidence intervals (CI) 1.9 to 4.6). There was no significant difference in SMR between males and females. The SMR was 8.8 (95% CI 5.0 to 14) for the index cases and 0.96 (95% CI 0.3 to 2.3) for the non-index cases based on five deaths. The overall mean % predicted forced expiratory volume in one second (FEV1) at entry was 83% with a significant difference between index cases (54%) and non-index cases (100%) (p < 0.001). The difference in the ratio of FEV1 to forced vital capacity (FVC) was also highly significant with values of 0.57 and 0.79 for index and non-index cases, respectively (p < 0.001). In the non-index group only three had an FEV1% predicted of less than 70%. CONCLUSIONS: Occupational exposure to airway irritants did not have any significant influence on the development of emphysema. Only a few life time non-smokers develop severe emphysema; most never develop pulmonary symptoms and thus remain undetected unless family members of index cases are screened. PMID- 9741369 TI - Effects of theophylline and ipratropium bromide on exercise performance in patients with stable chronic obstructive pulmonary disease. AB - BACKGROUND: The effects of theophylline or anticholinergic agents on exercise capacity in patients with chronic obstructive pulmonary disease (COPD) remain controversial. The aim of the present study was to compare the effect of an oral theophylline with an inhaled anticholinergic agent and to examine the effects of combined therapy on exercise performance using progressive cycle ergometry. METHODS: Twenty one men with stable COPD and a mean (SD) forced expiratory volume in one second (FEV1) of 1.00 (0.40) 1 were studied. Theophylline (600 or 800 mg daily), ipratropium bromide (160 micrograms), a combination of both drugs, and placebo were given in a randomised, double blind, four period crossover design study. Spirometric data, pulse rate, and blood pressure were assessed before and at 90 and 120 minutes after inhalation. Symptom limited progressive cycle ergometer exercise tests (20 watts/min) were performed 90 minutes after each inhalation, and dyspnoea was measured during exercise using the Borg scale. RESULTS: The mean (SD) serum theophylline concentration was 18.3 (6.3) micrograms/ml, and seven patients had side effects during treatment with theophylline. Theophylline and ipratropium bromide produced greater increases in FEV1, maximal oxygen consumption, maximal minute ventilation, and several dyspnoea ratios than placebo. There were no differences between theophylline and ipratropium bromide except in maximal heart rate. A combination of both drugs produced greater improvements in pulmonary function and exercise capacity than either drug alone. CONCLUSIONS: Both high dose theophylline and high dose ipratropium bromide improved exercise capacity in patients with stable COPD. Although data based on short term effects cannot be directly applied to long term therapy, theophylline added to an inhaled anticholinergic agent may have beneficial effects on exercise capacity in patients with COPD. PMID- 9741371 TI - Medium term results of lung transplantation for end stage pulmonary sarcoidosis. AB - BACKGROUND: Lung transplantation is an accepted therapeutic option for patients with end stage pulmonary sarcoidosis. However, the medium term outcome of transplantation in this patient group is unknown. METHODS: This study was performed to evaluate our experience with lung transplantation for end stage pulmonary sarcoidosis. Between July 1988 and July 1997 12 patients (nine men) underwent lung transplantation for sarcoidosis at our institution. Ten underwent single lung transplantation and two double lung transplantation. RESULTS: Survival at three and five years was 70% and 56%, respectively. Three patients developed obliterative bronchiolitis at six, 18, and 45 months. One died at the time of retransplantation. Sarcoid granulomas have recurred in the donor organ in three patients. In one the development of granulomas has been associated with clinical deterioration, necessitating retransplantation. Mean (SD) forced expiratory volumes in one second at three and five years were 1.37 (0.67) 1 and 1.34 (0.13) 1, respectively. CONCLUSIONS: Lung transplantation is a viable option for patients with end stage pulmonary sarcoidosis. The medium term results are comparable with patients undergoing lung transplantation for other indications. Despite histological recurrence of sarcoidosis, the risk of clinically important recurrence is low. PMID- 9741370 TI - Clinical profiles of Chinese patients with diffuse panbronchiolitis. AB - BACKGROUND: Diffuse panbronchiolitis (DPB), characterised by progressive sinobronchial sepsis, is well characterised in Japanese subjects but not in other ethnic groups. The experience with DPB in seven Chinese patients is described and the clinical profiles compared with those of Japanese subjects. METHODS: Seven Chinese patients (three women; mean (SD) age 48(18.6) years, all never smokers) who attended a teaching hospital centre and fulfilled the diagnostic criteria for DPB were assessed prospectively for clinical, radiological, lung function, microbiological, and other "characteristic" laboratory parameters. RESULTS: Lung function assessment showed a typical obstructive pattern (n = 5) and air trapping (n = 7). Typical bronchiolar infiltration by lymphocytes and plasma cells and accumulation of foamy macrophages in the intraluminal tissue were detected in open lung biopsy specimens (n = 2). Chest radiographs and high resolution computed tomographic scans revealed hyperinflation, diffuse nodules, bronchial thickening and dilatation, peripheral hypoattenuation, and bronchiolectasis. Radiological improvement, manifest as a reduction in nodular density and bronchial thickening, and persistence of other abnormalities such as air trapping were not accurately depicted by the classical Nakata or Akira classifications. The other "characteristic" features such as HLA-B54, IgG subclass deficiency, raised CD4/CD8 T lymphocyte ratio, cold haemagglutinaemia, raised IgA, IgG, and rheumatoid factor were not present. Treatment with erythromycin led to excellent responses in symptoms, lung function indices, and the radiological picture. A review of the non-Japanese cases in the literature reveals that this absence of typical "additional features" in DPB might also be applicable to non-Japanese patients. CONCLUSIONS: We report the only series of non-Japanese Mongoloid patients with well characterised DPB who had uncharacteristic investigation profiles. This experience should help other clinicians in the investigation and management of DPB in non-Japanese patients. PMID- 9741372 TI - Totally implantable venous access devices in children with cystic fibrosis: incidence and type of complications. AB - BACKGROUND: Totally implantable vascular access devices (TIVADs) are accepted as a safe and effective method of facilitating long term intravenous therapy. We report our experience of the use of these devices in children with cystic fibrosis with a particular focus on the incidence and type of complications. METHODS: The medical records of patients with cystic fibrosis who underwent placement of a TIVAD at the Royal Children's Hospital, Melbourne, Australia from January 1987 to October 1996 were reviewed. Venous ultrasonography with Doppler was performed in surviving patients with a TIVAD in situ from November 1996 to April 1997 to detect occult thrombotic complications. RESULTS: A total of 57 TIVADs were implanted in 44 children with a median functional duration of 700 days (range 27-3347 days). Twenty one children had devices inserted without complications. Forty eight complications (30 mechanical, 18 infectious) occurred in 36 devices in 23 children during a total functional duration of 53,057 catheter days. Mechanical complications occurred in 53% of devices (one per 1712 catheter days). Symptomatic venous thrombosis occurred five times in four patients (9%). Infectious complications occurred in 32% (one per 2948 catheter days) while sepsis occurred in five devices (9%). Doppler ultrasonography detected unsuspected thrombosis in two of 10 patients examined. CONCLUSIONS: While TIVADs provided effective long term intravenous access, septic and thrombotic complications caused significant morbidity in this population. Careful patient selection, adherence to aseptic technique for access and blood sampling, and periodic ultrasonography with Doppler to detect early thrombosis may help reduce these risks. PMID- 9741373 TI - Comparison of nose and face mask CPAP therapy for sleep apnoea. AB - BACKGROUND: Many patients with sleep apnoea/hypopnoea syndrome (SAHS) find nasal continuous positive airway pressure (CPAP) treatment unsatisfactory due to side effects related to mouth air leakage. A study was performed to compare side effects with face mask and nose mask CPAP therapy in patients with SAHS, with and without uvulopalatopharyngoplasty (U3P). METHODS: Twenty newly diagnosed patients with SAHS took part in a randomised double limb trial of face or nose mask CPAP therapy (four weeks per limb) in which CPAP compliance in terms of machine run time was measured and patients answered a symptom questionnaire on side effects resulting from the mask. Ten patients with SAHS with U3P (SAHS/U3P) who were already regular users of nasal CPAP were also given a four week trial of face mask CPAP to compare compliance and symptoms. Ten patients with SAHS were matched with the 10 SAHS/U3P patients for body mass index, age, apnoea/hypopnoea index, and CPAP pressure. Long term compliance was estimated one year after the mask comparison studies. RESULTS: For patients with SAHS nightly compliance was higher with a nose mask (mean (SE) 5.3 (0.4) hours/night CPAP) than with a face mask (4.3 (0.5) hours/night CPAP), p = 0.01 (mean difference 1.0 hour/night, 95% CI 1.8 to 0.3). Nose masks were rated more comfortable by 19 of 20 patients (p < 0.001) despite more mouth leak related symptoms. For SAHS/U3P patients compliance was marginally higher with nose masks (5.1 (0.7) hours/night CPAP) than with face masks (4.0 (0.8) hours/night CPAP), p = 0.07 (mean difference 1.1 hour/night, 95% CI 2.1 to 0.1). Nose masks were rated more comfortable by seven of 10 patients. There were no significant differences in side effect scores with face and nose masks. At one year nine of 10 SAHS patients and nine of 10 SAHS/U3P patients were still using CPAP. Compliance was 5.4 (0.6) hours/night for the SAHS patients and 3.5 (0.4) hours/night for the SAHS/U3P patients, p = 0.02 (mean difference 1.9 hour/night, 95% CI 3.6 to 0.3). CONCLUSIONS: Compliance is greater with nose mask CPAP than with face mask CPAP because the overall comfort is better and compensates for increased symptoms associated with mouth leakage. Improved face mask design is needed. PMID- 9741374 TI - Reduced mortality in association with the acute respiratory distress syndrome (ARDS). AB - BACKGROUND: A study was undertaken to investigate possible reductions in mortality and/or changes in outcome predictive factors in patients with the acute respiratory distress syndrome (ARDS) managed in a single centre. METHODS: The study was a prospective observational cohort study of two patient populations with ARDS. Group 1 comprised 41 patients enrolled between May 1990 and April 1993, and group 2 consisted of 78 patients enrolled between June 1993 and March 1997. The end points of the study were mortality and various factors predictive of death. RESULTS: There was a marked reduction in mortality between groups 1 and 2 (66% versus 34%; relative risk 1.77; CI 1.23 to 2.55). There were no significant differences between the groups in terms of age (40.6 (3.3) versus 45.5 (2.2) years), APACHE score (14.5 (0.72) versus 13.6 (0.1)), lung injury score (2.95 (0.07) versus 2.8 (0.1)), incidence of multi-organ failure (29% versus 32%), incidence of sepsis (31% versus 39%), or PaO2/FIO2 (kPa) ratio (11.8 (0.67) versus 12.0 (0.6)). There was a significantly lower proportion of men in group 1 (51% versus 74%). The case mix of the two groups was closely matched: following elective surgery 48% versus 48%, trauma 17% versus 16%, primary lung injury 12% versus 24%. Patients in group 1 were supported using several ventilatory and other modes (volume preset, non-inverse ratio ventilation, n = 15; pressure controlled inverse ratio ventilation (PC-IRV), n = 11; ultra high frequency jet ventilation (UHFJV), n = 13; an intravascular oxygenation device (IVOX) and extracorporeal gas exchange (ECGE), n = 2). Within group 1 no significant difference in mortality was observed between the patients on volume controlled ventilation and the remainder. In group 2 all patients received PC-IRV (n = 78) but, in addition, some received other support techniques (UHFJV n = 4, ECGE n = 2). In group 1 only sepsis on admission (21% (survivors) versus 56% (non survivors)) predicted death. In group 2 age of survivors and non-survivors (41.2 (2.6) versus 52.6 (3.5)), APACHE score (12.2 (0.6) versus 15.8 (0.9)), and PaO2/FIO2 (12.8 (0.86) versus 10.5 (0.72)) predicted survival, but not the incidence of sepsis or multi-organ failure. CONCLUSIONS: In recent years a highly significant reduction in mortality associated with ARDS has been observed between two groups of patients well matched for disease severity and case mix. Changes in ICU organisation rather than specific interventions may account for this reduction, although different ventilatory and other management strategies used in the two groups may also be relevant. PMID- 9741375 TI - Parental smoking, bronchial reactivity and peak flow variability in children. AB - BACKGROUND: A systematic quantitative review was conducted of the evidence relating environmental tobacco smoke to bronchial hyperresponsiveness (BHR) during childhood. METHODS: Twenty-nine relevant studies were identified after consideration of 1593 articles selected by electronic search of the Embase and Medline databases using keywords relevant to passive smoking in children. The search was completed in April 1997. RESULTS: Of 19 studies using challenge tests in children of school age, 10 (5759 children) could be summarised as the odds ratio of being bronchial hyperreactive in children exposed to environmental tobacco smoke compared with those not exposed. The pooled odds ratio for maternal smoking was 1.29 (95% confidence limits 1.10 to 1.50) with no evidence of heterogeneity between studies. However, in five further studies of 3531 children providing some evidence, but not odds ratios, none were statistically significant. A further four studies on 5233 children have collected data but are not published. In contrast, all four studies of circadian variation in peak expiratory flow found increased variation in children exposed to environmental tobacco smoke. CONCLUSIONS: A clear effect of exposure to environmental tobacco smoke on BHR in the general population has not been established. While the meta analysis suggests a small but real increase in BHR in school aged children, it seems likely that this estimate is biased upwards due to publication bias. In contrast, limited evidence suggests greater variation in peak expiratory flow in children of smoking parents. PMID- 9741376 TI - Pathogenesis of lower respiratory tract infections due to Chlamydia, Mycoplasma, Legionella and viruses. PMID- 9741377 TI - High resolution computed tomography in asthma. PMID- 9741379 TI - Commentary: "histiocytosis X". PMID- 9741378 TI - Brittle asthma. AB - We believe that the asthma phenotypes we have defined as types 1 and 2 brittle asthma appear to be defined subgroups of asthma. For example, we have characterised patients with type 1 brittle asthma, as defined in this review, on the basis of peak flow variability and treatment and these patients remain a separate group when assessed by other means such as psychosocial factors, immunoglobulin levels, and atopy. The question remains as to whether they are truly separate groups with entirely different pathogenetic influences or whether they simply represent the severe end of the spectrum. Our suggested classification into types 1 and 2 forms a useful start for studies of this condition, although prospective evaluation of patients with severe asthma is the only way of substantiating the validity of these definitions which will then enable investigation of possible mechanisms. However, these patients are rare and in order to study them as a group a national register would need to be set up along the lines of the West Midlands Brittle Asthma Register, perhaps recruiting all "at risk" patients and then using this resource as a means of exploring the different asthma phenotypes within this broad grouping, including brittle asthma. PMID- 9741380 TI - Recurrence of recipient Langerhans' cell histiocytosis following bilateral lung transplantation. AB - Langerhans' cell histiocytosis may cause irreversible respiratory failure due to progressive destruction of lung parenchyma and widespread cystic change. Transplantation offers a therapeutic option. A case is described of recurrence of Langerhans' cell histiocytosis which was associated with deterioration in lung function four years following bilateral lung transplantation. Patients transplanted for Langerhans' cell histiocytosis should be followed up with this complication in mind. PMID- 9741381 TI - Recurrence of Langerhans' cell granulomatosis following lung transplantation. AB - A case is presented of pulmonary Langerhans' cell granulomatosis which recurred following lung transplantation and responded to cyclophosphamide. This suggests that the primary abnormality in this condition lies in the Langerhans' cell or precursor dendritic cell. PMID- 9741382 TI - Should steroids be first choice for asthma? PMID- 9741383 TI - The tobacco epidemic in Japan. PMID- 9741384 TI - Tobacco advertising ban in Europe. PMID- 9741385 TI - Nitrate and nitrite anion concentration in the intact cerebral cortex of preterm and nearterm fetal sheep: indirect index of in vivo nitric oxide formation. AB - Pregnant sheep with a microdialysis probe implanted in the fetal cerebral cortex were used to determine if nitrate and nitrite anions (nitrate/nitrite) could be quantitated in the microdialysate as an indirect index of in vivo nitric oxide formation. Pregnant ewes (term, about 147 days) were surgically instrumented at gestational day (GD) 90 (n = 3; preterm) and GD 121 (n = 3; nearterm). Three days later, following an overnight probe equilibration period, five dialysate samples were collected continuously on ice at 1-h intervals (infusion rate of 1 (microl/min). The nitrate/nitrite concentration was determined by reducing a 10 microl aliquot of each dialysate fraction with hot acidic vanadium followed by chemiluminescence quantitation of the nitric oxide product. The lower limit of quantitative sensitivity of the method is 25 picomoles. Nitrate/nitrite concentration was 16.6+/-7.3 microM for the preterm fetus and 19.7+/-1.9 microM for the nearterm fetus. The data demonstrate that nitrate/nitrite, as an index of in vivo nitric oxide formation, can be quantitated in microdialysate samples collected from the intact fetal sheep cerebral cortex. PMID- 9741386 TI - An alternative method to evaluate the nature of an antagonist and its potency: a theoretical approach. AB - The Schild analysis is certainly the most reliable method for antagonism studies. The Schild regression allows one to determine the parameter of the Schild-slope, which represents a powerful diagnostic tool when investigating the nature of an antagonist and, consequently, to evaluate its potency. Nevertheless, in functional pharmacology, often practical reasons lead the experimenter to obtain an inhibition curve for the antagonist and calculate its potency by means of equations, which can be considered as a derivation of the Cheng-Prusoff analysis. This approach is considered theoretically invalid, because it does not allow to know the exact nature of the antagonism, and thus the evaluation of the antagonist dissociation constant can be meaningless. In this paper, a new method is proposed, which, by means of an equation closely similar to the Schild one, permits one to obtain a linear regression analysis, giving a slope value absolutely equivalent to the Schild-slope. This method allows us to determine both the nature and the potency of an antagonist, and requires an experimental procedure substantially analogous to the one performed to obtain an inhibition curve. PMID- 9741387 TI - A novel method for chronic measurement of pleural pressure in conscious rats. AB - Pleural pressures are used to evaluate lung function and are generally measured acutely in anesthetized animals. Previous attempts to measure pleural pressure chronically in conscious animals have involved surgical implantation of pressure sensitive catheters directly into the pleural cavity. The success of these techniques has been limited by lung damage and/or tissue growth and encapsulation of the pressure-sensitive catheter with damping or loss of the signal. These problems have been eliminated by developing a novel surgical procedure for placement of a pressure-sensitive catheter beneath the pleural surface. The catheter (attached to a radiotelemetry transmitter) is surgically implanted beneath the serosal layer of the esophagus within the thoracic cavity. This is accomplished by making a small incision in the serosal layer of the esophagus caudal to the diaphragm and advancing the catheter cranially into the thoracic cavity until pressure changes are maximal. The accuracy of these measurements was verified by comparison with direct pleural pressure measurements over the range of -3 to -34 cm H2O. The pleural pressure changes remained constant for at least 14 weeks following surgery, and there was no evidence of tissue damage or growth around the catheter. This novel method for measuring pleural pressure chronically in conscious rats will facilitate evaluation of the effects of drugs, environmental agents, or disease on respiratory function by allowing repeated and simultaneous measurements of both ventilatory (breathing) patterns and lung function in conscious animals. PMID- 9741388 TI - In vitro correlation between two colorimetric assays and the pyruvic acid consumption by fibroblasts cultured to determine the sodium laurylsulfate cytotoxicity. AB - The target of this research was to determine the cytotoxicity of sodium laurylsulfate on single-layer cultures of human fibroblasts, using two colorimetric methods (neutral red and MTT tests) and the evaluation of the pyruvic acid consumption by the cells. For the determination of the cytotoxicity by colorimetric tests, we have determined the absorbance at 540 nm using a spectrophotometer. Pyruvic acid, present in the culture medium, is the mitochondria's C3 energetic metabolite. So, a measure of the cell's consumption of pyruvic acid was developed. The reaction is as follows: Pyruvic acid + NADH - > Lactic acid + NAD+ and the enzyme employed is the LDH (lactate dehydrogenase). This method can be used to measure cytotoxicity, proliferation, and the cell's activation. The method is rapid, precise, and lacks any toxic byproduct. The absorbance was measured using a spectrophotometer at 340 nm. The consumption of pyruvic acid follows upon the fibroblast's growth. Sodium laurylsulfate cytotoxicity test after 24 h shows that the NR colorimetric test and the pyruvic acid consumption are correctly correlated (r = 0.91, alpha = 0.05). This dosage can be used to study the barrier properties of the corneocyte layer without destroying the artificial skin. PMID- 9741389 TI - Lipoteichoic acid induces preterm delivery in mice. AB - The purpose of this study was to determine whether or not lipoteichoic acid (LTA) could induce preterm delivery in mice. On days 15 and 17 of pregnancy, female C3H/HeN mice impregnated by male B6D2F1 mice were given intraperitoneal injections of LTA (12.5-75 mg/kg, single dose or repeated doses at a 3-h interval). We examined the changes in cervix, placental trophoblasts, and plasma and amniotic fluid concentrations of interleukin-1alpha (IL-1alpha), interleukin 6 (IL-6), and tumor necrosis factor-alpha (TNF-alpha) after dosing with LTA. In addition, the effect of LTA on the contraction of isolated uterine muscle from pregnant mice was also measured. The incidence of preterm delivery was highest (100%), when the pregnant animals were treated with 75 mg/kg LTA twice on day 15 of pregnancy or with 25 mg/kg LTA twice on day 17 of pregnancy. LTA-accelerated cervical ripening and placental abruption preceding the onset of preterm delivery, as well as increased plasma and amniotic fluid concentrations of IL 1alpha, IL-6, and TNF-alpha. Also, LTA increased contraction of uterine muscle strips. In conclusion, LTA induced preterm delivery in mice in the same manner as lipopolysaccharide (LPS), but the effective dose of LTA was larger than that of LPS. PMID- 9741390 TI - A reliable procedure for comparison of antioxidants in rat brain homogenates. AB - Lipid peroxidation is a major consequence of oxidative stress and an important cause of neuronal damage in ischaemic injuries and neurodegenerative disorders such as Parkinson's disease. Recent research has focused on the development of antioxidant drugs which may delay or minimize neurodegeneration. Rapid and reliable assays are therefore necessary in order to evaluate novel antioxidant compounds. A widely adopted method for measurement of lipid peroxidation is the thiobarbituric acid reacting substances (TBARS) assay. Several variations of this method have appeared in the literature, some of which have been tested by us without success. We have therefore established a reliable procedure which takes into account the most important factors previously found to influence the TBARS method. Briefly, various concentrations of drug were added to rat brain homogenates (10% w/v in 20 mM Tris-HCl buffer, pH 7.4) and incubated at 37 degrees C for 10 min before addition of ammonium ferric sulphate (100 or 1000 microM) and a further incubation at 37 degrees C for 30 min. Proteins were then precipitated with 8.1% sodium dodecyl sulphate, the reaction stopped with 20% acetic acid, and the samples were then centrifuged for 15 min. Aliquots of supernatant were added to an equal volume of thiobarbituric acid (0.8%), samples were heated at 95 degrees C for 30 min, and then cooled on ice before reading at 532 nm. The present adaptation represents a simple and highly reproducible assay which does not require difficult extraction procedures with hazardous chemicals and results in a stable chromagen. The method has been evaluated using a number of structurally distinct antioxidants and iron chelators. IC50 values (microM) for percentage inhibition of TBARS formation were as follows: desferroxamine (1.1), U83836E (1.7), butylated hydroxytoluene (13), U74500A (20), LY231617 (22), idebenone (89), and Trolox (110). This order of potency was comparable to that found with a commercially available, but expensive kit designed to specifically measure malondialdehyde (Spearman's rank correlation coefficient, p < 0.01). PMID- 9741391 TI - Measurement of bronchoconstriction using whole-body plethysmograph: comparison of freely moving versus restrained guinea pigs. AB - We have previously measured pulmonary function in guinea pigs using a double chambered plethysmograph, however, the question remains regarding the accuracy of the double-chamber to gauge the long-term pulmonary function of late asthmatic response. This may be affected by confounding factors, such as stress on the animal and differences in size of the collar around the neck. Therefore, in this study we compared histamine-induced bronchoconstriction in the same guinea pigs using a single- versus a double-chambered body box. In the double-chambered body box, the specific airway resistance is proportional to time delay between thoracic and nasal flows and measured in cmH2O x s. Whereas, in the single chambered body box, PenH units (Enhanced Pause) reflect "effort of breathing." This is measured as the pause between inspiration and expiration. Doubling concentrations of histamine (12.5-200 microg/ml dissolved in normal saline) were administered by DeVilbiss nebulizer for 1 min, followed by 1 min suction of residual drug in the chamber, and then the airway resistance was recorded by the computer for the following 3 min. There was a 15-min wash-out period between two doses of histamine. There was no statistically significant difference (p > 0.05) in the PC100 values for histamine between the two methods, however, it was much easier to work with the single-chambered body box in terms of handling the animal and eliminating the possible influence of collar placement on the bronchoconstriction. In conclusion, the data suggests histamine challenges produce equivalent PC100 data in both the double-chambered plethysmograph with sRAW units and single-chambered plethysmograph using the PenH units. PMID- 9741392 TI - Improved myeloperoxidase assay for quantitation of neutrophil influx in a rat model of endotoxin-induced uveitis. AB - Previously described models of endotoxin-induced uveitis quantify neutrophil influx into the eye using biochemical or direct cell count methods that result in an underestimation of ocular leukocyte accumulation following the inflammatory stimulus. We have optimized the rat model of endotoxin-induced uveitis by first overcoming interference in the biochemical assay of myeloperoxidase due to endogenous ocular reductants and cellular constituents containing free thiol functional groups. This was accomplished by simultaneously 1) extensively diluting soluble, interfering substances and 2) blocking tissue sulfhydril functional groups during tissue homogenization. Uveitis was induced in rats by subplantar injection of endotoxin. Twenty-four hours later, eyes were enucleated, homogenized, fractionated, and myeloperoxidase activity of neutrophils sedimenting with the membranous pellet was extracted. Previously published extraction procedures yielded only 40% of total assayable myeloperoxidase activity. Optimal recovery of myeloperoxidase activity (>twofold increase) was achieved only with two sequential extractions using 50 mM phosphate buffer (pH 7.4) containing 10 mM N-ethylmaleimide, and subsequent solubilization of myeloperoxidase activity by extraction with 0.5% hexadecyltrimethylammonium bromide in 50 mM phosphate buffer (pH 6.0). This modified extraction procedure and optimized myeloperoxidase assay conditions (300 microM hydrogen peroxide and 1.5 mM o-dianisidine) were then used to enhance the uveitis model. Maximum ocular neutrophil accumulation was observed at endotoxin doses of 100-200 microg. Total ocular neutrophil infiltrations ranged from 250,000 to 800,000 cells/globe. This leukocyte influx was inhibited dose-dependently by topical ocular administration of dexamethasone, with half-maximal inhibition observed at a concentration of 0.01%, w/v. Further validated by the correlation of biochemical results with histological evaluation, the refined methodology described in this report has application in assessing the ophthalmic therapeutic potential of antiinflammatory agents. PMID- 9741393 TI - Ferrous iron-induced luminol chemiluminescence: a method for hydroxyl radical study. AB - We have investigated the chemiluminescence signal of the ferrous iron in the presence of the luminol and lucigenin. Ferrous, but not ferric, iron produced a transient signal in the presence of luminol, but not lucigenin. Ferrous iron induced luminol chemiluminescence was significantly inhibited in a concentration dependent manner by superoxide dismutase (SOD) and catalase. Specific hydroxyl radical scavengers, mannitol and dimethyl sulfoxide (DMSO), also markedly attenuated the ferrous iron-induced chemiluminescence. Additionally, antioxidants, urate, ascorbate, and methionine produced concentration-dependent significant inhibitions in this chemiluminescence. These results show that the hydroxyl radical generation is dependent on simultaneous formation of superoxide and hydrogen peroxide (H2O2). Ferrous iron does not generate a chemiluminescence signal in the presence of lucigenin suggesting that the formation of a hydroxyl radical is responsible for the luminol chemiluminescence. Thus, the present study has established a simple and inexpensive cell-free screening method for monitoring the scavenging effects of drugs on the hydroxyl radical. PMID- 9741394 TI - Oxidative damage to nucleic acids in motor neurons containing mercury. AB - Heavy metals have been implicated in the pathogenesis of sporadic motor neuron disease (MND). We were interested to see if inorganic mercury leads to oxidative damage in motor neurons since free radicals have been suspected to be involved in MND, so a method to examine oxidatively-damaged DNA in situ was used to examine individual motor neurons. Mice were exposed to 500 microg/m3 of mercury vapour for 2 h. Two, five, or ten days later sections from formalin-fixed, paraffin embedded blocks of cervical spinal cord were incubated in avidin-FITC. Sections were examined under a fluorescence microscope and photographs of pairs of mercury exposed and control spinal motor neurons were analysed semi-quantitatively for the amount of fluorescence using an image analysis program. Avidin fluorescence was seen in the perikaryon of both control and mercury-exposed motor neurons. In each control-mercury pair (four pairs per group) significantly more perikaryal fluorescence was seen in mercury-containing than in control motor neurons (Mann Whitney testing). Mercury within the motor neuron perikaryon therefore leads to increased avidin binding, an indicator of oxidative damage to DNA. The findings support the hypothesis that an environmental toxin such as mercury can enter and damage motor neurons. PMID- 9741395 TI - Role of the leukocyte-adhesion molecule L-selectin in experimental autoimmune encephalomyelitis. AB - L-selectin is an adhesion molecule expressed on T cells and monocytes. It mediates rolling--the initial step of transendothelial migration. In this study, we investigated the role of L-selectin in the pathogenesis of experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis. EAE was induced in Lewis rats by active sensitization with myelin basic protein (MBP EAE), or by adoptive transfer using MBP specific T cells (AT-EAE). Treatment with HRL3, a monoclonal antibody to L-selectin, and its F(ab')2 fragments efficiently suppressed MBP-EAE, and had a mild inhibitory effect on AT-EAE. Histological examination revealed a marked reduction of inflammatory infiltrates after treatment with HRL3. Administration of the control antibody HRL4 did not significantly alter the course of the disease. HRL3 caused T-cell depletion in the draining lymph nodes and spleen and a downregulation of L-selectin expression on T cells. We conclude that L-selectin-dependent mechanisms are involved in the pathogenesis of EAE. Modulation of L-selectin in vivo by antibodies or by competitive antagonists could be a novel therapeutic approach to autoimmune diseases of the central nervous system. PMID- 9741396 TI - Sustained release dosage of thyrotropin-releasing hormone improves experimental Japanese encephalitis virus-induced parkinsonism in rats. AB - Thyrotropin-releasing hormone (TRH) has been reported to have some possibilities toward the treatment of affective CNS disorders. However, long term treatments with daily injections are often required. Effects of TRH-SR (sustained release microspheres of TRH) which is encapsulated in copoly (dl-lactic/glycolic acid) using an in-water drying method were investigated in experimental Japanese encephalitis virus (JEV)-induced post-encephalitic parkinsonism rats by a pole test and high performance liquid chromatography (HPLC) with an electrochemical detector (ECD). We have already reported that in adult Fischer rats killed 12 weeks after infection with JEV at the age of 13 days a marked decrease of tyrosine hydroxylase-positive neurons was found in the bilateral substantia nigra. TRH-SR (3 mg/kg per 2 weeks, 4 times injections, subcutaneous [s.c.]) improved bradykinesia observed in the JEV-induced parkinsonism rats. Dopamine (DA) concentrations in the JEV-infected rats were profoundly reduced in the striatum as compared with controls. TRH-SR (3 mg/kg, once, s.c.) increased DA in the striatum 7 days after the injection. Although the pathomechanism of post encephalitic parkinsonism is different from that of Parkinson's disease and TRH possesses a variety of CNS effects as well, these results suggest that TRH-SR play a possible role in the treatment of Parkinson's disease in addition to post encephalitic parkinsonism as a supportive drug of L-DOPA. PMID- 9741397 TI - The value of cerebrospinal fluid antiviral antibody in the diagnosis of neurologic disease produced by varicella zoster virus. AB - We studied four patients with subacute to chronic varicella zoster virus (VZV) infection of the central nervous system (CNS). VZV infection was verified by detecting antibody to VZV in the cerebrospinal fluid (CSF). VZV caused myelitis in two patients and encephalitis in two patients. In one of the patients with VZV encephalitis, in addition to VZV IgM antibody, VZV DNA was found in the CSF. Among the four patients with VZV infection of the CNS, CSF antibody to VZV was the crucial diagnostic laboratory test which corroborated the clinical features, and indicated that VZV caused neurologic disease. In addition to looking for amplifiable VZV DNA in the CSF of patients with neurologic disease whose clinical and radiologic features suggest VZV infection, we also recommend a search for CSF antibody to VZV, particularly in patients with intervals of weeks to months between zoster and the onset of neurologic disease, or in those patients without rash in whom the tempo of illness is unknown. PMID- 9741398 TI - Consensus opinion on diagnosis of cerebral circulatory arrest using Doppler sonography: Task Force Group on cerebral death of the Neurosonology Research Group of the World Federation of Neurology. AB - BACKGROUND AND PURPOSE: Oscillating flow or systolic spikes are typical Doppler sonographic flow signals found in the presence of cerebral circulatory arrest, which if irreversible, results in brain death. The Neurosonology Research Group (NSRG) of the World Federation of Neurology (WFN) created a Task Force Group in order to evaluate the role of Doppler-sonography as a confirmatory test for determining brain death. METHODS: The available evidence from the literature has been reviewed and discussed by a group of experts, the members of the Task Force Group on cerebral death of the NSRG. RESULTS AND CONCLUSIONS: Extra- and intracranial Doppler-sonography is a useful confirmatory test to establish irreversibility of cerebral circulatory arrest as optional part of a brain death protocol. Doppler-sonography is of special value when the therapeutic use of sedative drugs renders electroencephalography unreliable. Doppler-sonographic criteria are defined and guidelines for the use of Doppler-sonography in this setting are presented. PMID- 9741399 TI - Prognosis of epilepsy associated with single CT enhancing lesion: a long term follow up study. AB - This is a retrospective analysis to study the long term prognosis of epilepsy associated with single CT enhancing lesion (SCTEL). Follow up CT scan showed resolution of the lesion in all of the 102 patients. Seizures did not recur in 64 (63%) patients after starting antiepileptic drugs. Twenty eight (27.5%) patients had recurrence of seizures for a median period of 2 months before remission was achieved. In the remaining ten (10%) patients seizures remitted only after albendazole therapy and the median period of seizure recurrence was 8 months. Sixteen (42%) of the 38 patients who had recurrence of seizures had type B CT lesion (ring lesion with central enhancing area, probably scolex) (P<0.02 (95% CI 3.2-40.3)). Patients with type B CT lesion had more numbers of seizures and also longer intervals between first and last seizure. Antiepileptic drugs were withdrawn in all the 102 patients. The mean period of follow up was 45 months (range 19-101). Only one patient had a relapse and his follow up CT showed gliotic scar at the site of the previous lesion. We conclude that epilepsy associated with SCTEL is a benign form of epilepsy and seizures recur as long as the lesion persists. Antiepileptic drugs can safely be withdrawn once the follow up CT shows resolution of the lesion. PMID- 9741400 TI - Progressive aphemia in a patient with Pick's disease: a neuropsychological and anatomic study. AB - We describe a patient with progressive aphemia with agrammatism that was later overlaid with buccofacial apraxia and pseudobulbar palsy. Pathological findings were consistent with those of classic Pick's disease with argyrophilic inclusions and neuronal achromasia, except for restricted cortical atrophy in the frontal operculum posterior to the pars opercularis (Brodmann Area 44). In addition, major neuronal loss was confined to the premotor cortex and the anterior half of the precentral gyrus (Area 6), which apparently explained the aphemia. The present case demonstrated that classic Pick's disease can show quite limited cortical atrophy in a patient who clinically presents with progressive aphemia. Also, our patient differed from the progressive non-fluent aphasia patients reported as having Pick's disease, who were all Pick variants, revealing that classic Pick's disease can be included in the spectrum of progressive aphasia syndrome. PMID- 9741401 TI - Magnetization transfer ratios of multiple sclerosis lesions with variable durations of enhancement. AB - We evaluated whether new multiple sclerosis (MS) lesions with variable durations of enhancement have different magnetization transfer ratios (MTR) at the time of their appearance. We scanned ten patients with relapsing-remitting MS every four weeks on four occasions. During each of the monthly sessions, we obtained dual echo, MT and T1-weighted scans. Five minutes after gadolinium-DTPA (Gd) injection, another T1-weighted scan was obtained. We measured the MTR of new enhancing lesions on co-registered quantitative MTR images. During the three month follow up, 54 new enhancing lesions were seen with a mean MTR value of 33.6% (SD=7.8%). The mean MTR values were 35.3% (SD=7.0%) for lesions enhancing on only one scan and 29.3% (SD=8.6%) for those enhancing on at least two consecutive scans (P=0.01). These results suggest that enhancing lesions in MS have heterogeneous pathological substrates, which may be associated with different durations of the enhancing phase. PMID- 9741402 TI - The first component of polyphasic motor evoked potentials is resistant to suppression by paired transcranial magnetic stimulation in humans. AB - The suppressive effect of paired transcranial magnetic stimulation on the different components of motor evoked potentials (MEPs) from the left first dorsal interosseous muscle (FDI) was investigated in ten healthy human subjects. Conditioning stimuli were delivered via a round coil centered on the vertex, and test stimuli via a figure-eight coil over the right motor area. The threshold stimulus intensity of the minimally contracted FDI was determined with each coil. The stimulus intensities of conditioning and test stimuli were 80% and 130-140% of the threshold intensity, respectively. The coil current direction at the intersection of the figure-eight coil was medio-lateral. The coil position for test stimuli was adjusted to evoke discretely polyphasic MEPs in the contracted FDI. Conditioning stimuli were followed by test stimuli at intervals of 1-10, and 15 ms. When the interstimulus intervals were less than 5 ms, the second negative phase of MEPs was suppressed compared to that of the unconditioned responses, whereas the first phase was not suppressed. The present results imply that the first phase reflects direct corticospinal volleys, which are resistant to cortico cortical inhibition. PMID- 9741403 TI - Single muscle fiber analysis in patients with 3243 mutation in mitochondrial DNA: comparison with the phenotype and the proportion of mutant genome. AB - An A-to-G point mutation at nucleotide pair (np) 3243 (3243 mutation) in mitochondrial DNA (mtDNA) is a well-known pathogenic mutation, which has been found in approximately 80% of patients with mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS). It has been reported that the 3243 mutation also occurs in individuals with non-MELAS phenotypes. The reasons for the phenotypic heterogeneity of the 3243 mutation have not been clarified, although it may be closely related with mtDNA heteroplasmy and their differing proportions in different tissues. We examined the proportion of mutant DNA in muscle specimens at the cellular level using single fiber analysis in five patients with the 3243 mutation: three were diagnosed clinically as having MELAS and two had mitochondrial diabetes mellitus (MDM). In both phenotypes, ragged-red fibers (RRF) contained a higher percentage of mutant DNA (average 89.8%) than non-RRF (average 42.4%). On the other hand, the proportion of mutant DNA in non-RRF revealed a wider range than in RRF and the average was higher in MELAS patients (58.5+/-27.3%) than that in MDM patients (26.3+/-27.9%), which correlated with biochemical and morphological mitochondrial abnormalities in muscle. These findings may reflect the underlying mechanisms of tissue specificity in each phenotype. PMID- 9741404 TI - Lactate stress test in the diagnosis of mitochondrial myopathy. AB - The aim of the study was to determine the sensitivity and specificity of the lactate stress test in the detection of mitochondrial myopathies. Thirty one healthy subjects, 10 patients with non-mitochondrial myopathy and 26 patients with mitochondrial myopathy underwent lactate stress testing at a standardized workload of 30 W during 15 min on a bicycle ergometer. Lactate was determined before the exercise (R1), 5, 10, 15 min after starting the exercise (S5, S10, S15) and 15 min after finishing the exercise (R2). A result was interpreted as pathologic if more than two of the five lactate values were above the corresponding upper reference limits. The upper reference limits for the venous lactate at R1, S5, S10, S15 and R2 were 1.9, 2.0, 2.1, 2.0 and 1.7 mmol/l respectively. The lactate stress test was pathologic in 1/10 of the non mitochondrial myopathies and in 18/26 of the mitochondrial myopathies. The sensitivity of the lactate stress test was 69%. The specificity of the test was 90%. In conclusion, the lactate stress test proved to be helpful for evaluating the integrity of the oxidative metabolism in the majority of patients with mitochondrial myopathy. PMID- 9741405 TI - Low-dose weekly methotrexate for progressive neuropsychiatric manifestations in Behcet's disease. AB - The most serious central nervous system (CNS) manifestation in Behcet's disease is a slowly progressive dementia (progressive NB), which may ultimately lead to the deterioration of the personality of patients. An open trial was designed to investigate the efficacy of low dose weekly methotrexate (MTX) therapy for progressive NB. Six patients with Behcet's disease, whose neuropsychiatric manifestations were judged to be progressive (4 females and 2 males, aged 55.0+/ 8.2 years), were given oral MTX (7.5-12.5 mg/week) until the end of the 12-month trial. The clinical responses of the patients to MTX were judged by neuropsychiatric findings, intelligence test, brain MRI scans and cerebrospinal fluid (CSF) IL-6 levels. After the 12-month trial, CSF IL-6 levels were found to be significantly decreased. Accordingly, the neuropsychological manifestations as well as the findings on MRI scans and intelligence quotients were not significantly worsened after the trial. Three patients presented with mild liver dysfunction, which returned to normal by decreasing the dose of MTX. However, 6 months after discontinuation of MTX, all the six patients showed significant exacerbation of the manifestations as evidenced by a decrease in verbal intelligence quotients along with the marked elevation of CSF IL-6. These results suggest that low dose weekly MTX therapy might have a beneficial effect in the treatment of progressive NB, although a trial for a longer period would be necessary. PMID- 9741407 TI - Ubiquitin-immunohistochemical investigation of atypical Pick's disease without Pick bodies. AB - Six cases of atypical Pick's disease (PD) without Pick bodies (PB) were examined immunohistochemically. These cases showed severe neuronal loss with gliosis predominantly in the temporal cortices. Ubiquitin immunohistochemistry revealed ubiquitin-positive intraneuronal inclusions in the dentate gyrus and ubiquitin positive neurites in the cerebral cortex. In the dentate gyrus, the dendrites in the stratum moleculare as well as the intraneuronal inclusions in the granular cells were positively stained. Both structures were composed of ubiquitin positive ribosome-like granular components and a few filamentous components immunoelectron-microscopically. In the cerebral cortex, ubiquitin-positive neurites were distributed in layers II-IIIab and layers V-VI, and were considered to be the distal dendrites from the small neurons. The dendrites and perikarya of these neurons contained ubiquitin-positive components similar to those in the dentate gyrus. Some ubiquitin-positive neurites were also found in the hippocampal subiculum, amygdala and striatum. The results of this study suggest that the granular cells in the dentate gyrus and the small neurons in the cerebral cortex share common ubiquitin-related and ribosome-associated abnormalities in both the perikarya and dendrites, that the degeneration of the perforant pathway caused by the parahippocampal lesion participates in the ubiquitin related abnormalities in the granular cells, and that PD cases with and without PB have common affected neurons, as shown immunohistochemically. PMID- 9741406 TI - Clinical and electrophysiological findings in critical illness polyneuropathy. AB - Sixty two patients with critical illness polyneuropathy (CIP) were studied prospectively to determine the clinical and electrophysiological profile, to assess the prognostic value of respiratory electrophysiology in determining the duration of ventilation and to analyze the role of neuromuscular blocking agents (NMBA) and steroids. Limb motor and sensory nerve conductions, bilateral phrenic nerve onset latencies, bilateral diaphragmatic compound muscle action potentials (CMAP), unilateral diaphragmatic needle electromyography (EMG), limb muscle EMG, time on the ventilator, time in the intensive care unit (ICU), dosage of NMBA and steroids were analyzed in 62 patients. The diagnosis of CIP was made by clinical criteria, electrophysiological criteria and exclusion of any other condition suspicious of an axonal neuropathy. The results of phrenic nerve conduction studies and diaphragmatic EMG were compared to normal mean values in 25 healthy subjects. The most common finding in our study were reduced CMAPs and abnormal spontaneous activity in muscle, occuring in 100%. Forty per cent had reduced CMAPs but normal sensory nerve action potentials (SNAP). These patients had normal CK-levels and normal findings, unspecific changes, type 2 fibre atrophy or denervation atrophy on muscle biopsy. Seventy seven per cent of patients had abnormal diaphragmatic CMAPs and spontaneous activity in the diaphragm indicating denervation of the diaphragm is common in CIP. There was no statistically significant relationship to the dosage of NMBA and steroids, and the respiratory electrophysiological studies, duration of ventilation and stay in the ICU. PMID- 9741408 TI - Phenotype variation correlates with CAG repeat length in SCA2--a study of 28 Japanese patients. AB - Spinocerebellar ataxia-2 (SCA2) is an autosomal dominant ataxia caused by an abnormal CAG repeat expansion in a novel gene on chromosome 12q24.1. The size of the mutant allele is unstable during transmission, and correlates inversely with age at onset. We studied eight Japanese SCA2 families, including 28 patients, to assess the effect of repeat length on the phenotype features of SCA2. Frequencies of slow eye movements (SEM), reflex activity, dementia, choreiform movements, and axial tremor correlated significantly with CAG repeat size. Parkinsonism was seen in a man homozygote for SCA2 mutation. The clinical variety of SCA2 is apparently influenced by the size of the mutant allele, as is the case in other CAG repeat disorders. PMID- 9741409 TI - Multiple hemostatic abnormalities in young adults with activated protein C resistance and cerebral ischemia. AB - BACKGROUND: The relationship between activated protein C resistance (APCR) and arterial stroke is uncertain. It has been speculated that multiple inherited or acquired prothrombotic conditions may alter the hemostatic balance towards thrombotic events. METHODS: Case series from a University medical center. RESULTS: In a series of 28 Caucasian patients under age 55 with cerebral ischemic events, nine were found to have activated protein C resistance. Five of nine patients (56%) had additional hematologic abnormalities. Four patients had elevated anticardiolipin IgG antibodies. Other abnormalities identified included Type I protein S deficiency, sticky platelet syndrome, and a positive lupus anticoagulant. CONCLUSIONS: Activated protein C resistance is relatively common in young adults with cerebral ischemic events and may be accompanied by other hematologic abnormalities. The constellation of hemostatic abnormalities may impact the type of and intensity of the antithrombotic regimen. There are also implications for family members of affected patients. Finding evidence of APCR should not preclude a complete hemostatic evaluation in the young stroke patient. PMID- 9741410 TI - Profound cerebrospinal fluid pleocytosis and Froin's Syndrome secondary to widespread necrotizing vasculitis in an HIV-positive patient with varicella zoster virus encephalomyelitis. AB - Demonstration of the direct involvement of cranial blood vessels by varicella zoster virus (VZV) is facilitated by immunohistochemistry (IHC), in situ hybridization (ISH) and polymerase chain reaction (PCR) techniques. The extent to which an inflammatory vasculitis serves as the pathogenic mechanism for VZV encephalomyelitis (VZVE) is still, however, debated. Most VZVE patients are immunocompromised and show little inflammation, either pre-mortem in cerebrospinal fluid (CSF) or at autopsy. We describe an HIV-positive patient with a moderately depressed CD4 count (304) who presented with massively elevated CSF protein (1800 mg/dl), bloody CSF and pleocytosis (1300 white blood cells (WBC)/mm3). His CSF was positive for VZV DNA by PCR. He was treated with acyclovir and foscarnet, but died. At autopsy, an unusually widespread, inflammatory, transmural vasculitis caused by VZV affected meningeal vessels at virtually all brain stem and spinal cord levels, causing multiple subpial hemorrhages and necrosis. Virus DNA in multiple areas of brain, brainstem and spinal cord was readily revealed by PCR, but not by the presence of viral inclusions, IHC or ISH. This case, with a clinically confusing presentation for VZVE, illustrates the extensive, albeit infrequent, degree of necrotizing vasculitis and CSF abnormalities that VZV is capable of producing. Antiviral therapy may have inhibited VZV genome replication and subsequent antigen production, resulting in negative ISH and IHC studies, but generated increased VZV genomic fragments that were detectable by the more sensitive PCR technique. PMID- 9741411 TI - Normative values of vibratory perception in 530 children, juveniles and adults aged 3-79 years. AB - Impaired vibratory perception is an early and frequent finding in various neuropathies. Quantitative vibratory threshold assessment refines the diagnosis of neuropathies but is based on psychophysical techniques requiring patient cooperation. Large, age and sex matched normative data bases are needed to better identify abnormal vibratory perception. In this study vibratory perception was tested at the second metacarpal bone and above the first metatarsal bone of 530 children, juveniles and adults aged 3.3-79.2 years. Thresholds assessed with a 128 Hz graded Rydel-Seiffer tuning fork, TF, were compared to three Vibrameter values, the vibration perception thresholds, VPT, determined with increasing vibration stimuli, the vibration disappearance threshold, VDT, determined with decreasing supraliminal stimuli, and the vibration threshold VT which equals the mean of VPT and VDT. The influence of gender, age, body height, weight and skin temperature at the tested site on thresholds was studied. Retest reliability was tested in 73 children aged 3.3-6.9 years and in 20 volunteers aged 5.2-66.1 years who were also tested for the influence of pretest skin warming on thresholds and for differences between results of the left and right body side. TF, VPT, VDT, VT were closely correlated with each other (Spearman: -0.67 .05). For these 104 patients, 50% (52 of 104) had perineural invasion, of which 38.5% (20 of 52) had proven extracapsular extension. In our hands, transrectal ultrasound-directed and staging biopsies afford more substantive results than sextant biopsies for detecting extracapsular extension. For our cohort of sextant T1-T2 diagnosed cancer (n = 100), 27% were upstaged to T3-T4 by transrectal ultrasound-directed staging biopsy. Thus, transrectal ultrasound directed staging biopsy has the ability to diagnose unsuspected extracapsular extension and objectifies prognosis and choice of definitive treatment. PMID- 9741418 TI - The role of tumor biomarkers as predictors of serum PSA recurrence after radical prostatectomy. AB - Tumor biomarkers (p53, bcl-2, Ki-67, and RT-PCR) were reviewed in the literature for their ability to predict prostate-specific antigen (PSA) biological recurrence after radical prostatectomy. All of them are strongly associated with PSA recurrence on univariate analysis. p53 seems to be better than current predictors, such as stage, grade, and positive surgical margins on multivariable analysis; further studies need to confirm bcl-2 and Ki-67 as better predictors of PSA recurrence. However, most of these studies were performed on radical prostatectomy specimens and will need to be confirmed on the preoperative prostate biopsy. The RT-PCR assay was strongly correlated with PSA recurrence and was one of the two best preoperative PSA recurrence predictors with serum PSA. PMID- 9741419 TI - The role of computerized tomography, magnetic resonance imaging, bone scan, and monoclonal antibody nuclear scan for prognosis prediction in prostate cancer. AB - The single most important issue in determination of treatment options for prostate cancer is accurate assessment of disease extent. Some prediction of probability is afforded by algorithms of patient and tumor characteristics, but definitive detection of disease extension before this decision often remains difficult. This is the critical issue in the healthy 58-year-old man depicted with relatively high-grade, high-volume prostate cancer and a moderately low serum PSA relative to these characteristics. Any combination of choices for evaluation and treatment of this patient is likely to generate some controversy. This article discusses both the changing trends in treatment patterns, which place more emphasis on noninvasive staging and the limited value of conventional radiographic evaluation to detect small volume or microscopic disease. Recent advances in imaging techniques with magnetic resonance and radiolabeled monoclonal antibodies may provide more precise localization of prostate cancer in these clinical circumstances. The relative merits and limitations of the current and selected emerging imaging technology for prostate cancer detection are provided in this article. PMID- 9741420 TI - Assessment of outcome prediction models for localized prostate cancer in patients managed with external beam radiation therapy. AB - A comparison of the ability of all published proposed clinical staging systems to predict time to prostate-specific antigen (PSA) failure after external beam radiation therapy for clinically localized prostate cancer was performed using an independent radiation database. Cox regression multivariable analyses were used to assess the significance of the proposed staging systems to predict time to post-radiation PSA failure in 465 radiation managed patients. Significant staging systems identified using Cox regression were further tested using established comparative estimates to define the most predictive system. Both the Risk Score staging system and the staging system based on the calculated volume of prostate cancer (cV(Ca)) and PSA optimized the prediction of time to post-treatment PSA failure. The cV(Ca)-PSA system, however, provided a more clinically useful stratification of outcome. Many clinical staging systems for prostate cancer have been proposed. A single clinical staging system for patients with localized prostate cancer based on parameters obtained during the routine workup provided a statistically and clinically significant stratification of outcome after external beam radiation therapy. PMID- 9741421 TI - Multivariate models as predictors of pathological stage using Gleason score, clinical stage, and serum prostate-specific antigen. AB - The patient presented is a 58-year-old man with newly diagnosed prostate cancer who likely has at least a 10- to 15-year life expectancy. In staging this man's extent of disease, several preoperative clinical variables are provided to assess whether the patient has local disease before offering definitive surgical therapy. It has been demonstrated that the combination of several of these variables in a multivariate analysis has greater predictive power than any of these variables do alone. Multivariate analysis using clinical stage, prostate specific antigen (PSA), and Gleason score will provide both the physician and patient with 95% confidence intervals for determining the probability of having organ-confined disease, extracapsular penetration, seminal vesicle involvement, and lymphatic metastases. Several of the clinical variables given indicate that this patient has advanced disease, such as a PSA of 12 ng/mL, a stage T2b lesion, Gleason sum 7 disease in 2 of 3 cores from the right side associated with perineural invasion, and an additional Gleason sum 7 biopsy from the contralateral apex. For a man with these preoperative variables, there is a 13% chance of organ-confined disease, an 18% probability of seminal vesicle invasion, and a 17% chance of positive pelvic lymph nodes based on a nomogram constructed from multivariate analysis. Using this information, this man should be counseled that he has a high probability (87%) of extracapsular disease and a significant risk (15% to 20%) of having either seminal vesicle or lymph node involvement. Recognizing the risks and benefits of various forms of definitive therapy for prostate cancer, the patient has additional information to make an informed decision. PMID- 9741422 TI - Systematic biopsy-based staging of prostate cancer: scientific background, individual variables, combination of parameters, and current integrative models. AB - Staging of the prostate cancer remains the mainstay of treatment decisions and prognostication. Multiple staging tests, which are currently available, fail to provide optimal staging accuracy in the management of prostate cancer. Systemic biopsy can provide useful data regarding bilaterality, Gleason grade, cancer map, perineural infiltration, tumor volume, percentage cancer and microvessel density. We have reviewed existing literature on systematic biopsy in this review. We have further discussed the role of neural network technology in integrating this information. PMID- 9741423 TI - DNA vaccination for leishmaniasis. PMID- 9741424 TI - Development of human granulocyte-macrophage colony-stimulating factor-transfected tumor cell vaccines for the treatment of spontaneous canine cancer. AB - Cytokine gene-engineered tumor vaccines are currently an area of intense investigation in both basic research and clinical medicine. Our efforts to utilize tumor vaccines in an immunotherapeutic manner involve canines with spontaneous tumors. We hypothesized that canine tumor cells, transfected with human granulocyte-macrophage colony-stimulating factor (hGM-CSF) cDNA in a plasmid vector, would prove nontoxic following intradermal administration, generate biologically relevant levels of protein, effect local histological changes at the sites of vaccination, and create a systemic antitumor response. Sixteen tumor-bearing dogs were admitted to a study of ex vivo gene therapy. Tumor tissue was surgically removed, enzymatically and mechanically dissociated, irradiated, transfected, and intradermally injected back into the patients. The dogs were vaccinated with primary autologous tumor cells transfected with hGM-CSF or a reporter control gene. hGM-CSF protein was detected (0.07 to 14.15 ng/vaccination site) at 24 hr postinjection and dramatic histological changes were observed, characterized by neutrophil and macrophage infiltration at the sites of injection of hGM-CSF-transfected tumor cells. This was in stark contrast to the lesser neutrophilic and eosinophilic infiltrates found at control vaccination sites. Objective evidence of an antitumor response was observed in three animals. These data, in a large animal translational model of spontaneous tumors, demonstrate in vivo biological activity of hGM-CSF-transfected autologous tumor cell vaccines. PMID- 9741425 TI - A system for shuttling 200-kb BAC/PAC clones into human cells: stable extrachromosomal persistence and long-term ectopic gene activation. AB - A novel shuttle vector, pBH140, has been constructed that allows stable maintenance of large genomic inserts as human artificial episomal chromosomes (HAECs) in mammalian cells. The vector, essentially a hybrid BAC-HAEC, contains an F-based replication system as in a bacterial artificial chromosome (BAC) and the Epstein-Barr virus (EBV) latent origin of replication system, oriP, for replication in human cells. A 185-kb DNA insert containing the entire human beta globin locus, including its locus control region (LCR), was retrofitted into this vector. The resulting beta-globin BAC-HAEC clone, p148BH, was transfected into human cells and analyzed for episomal maintenance and expression of the beta globin gene. FISH revealed an association of the vector with different human chromosomes but no integration. The beta-globin BAC-HAECs were present at an average copy number of 11-15 per nucleus in the stably transformed human cells. After 1 year of continuous in vitro cultivation, the HAECs persisted as structurally intact 200-kb episomes. While no beta-globin transcription could be detected in the parental D98/Raji cells, correctly spliced RT-PCR products were produced at significant levels in long-term cultures of the BAC-HAEC-transduced cells. The wide availability of BAC and PAC libraries, the ease in manipulating cloned DNA in bacteria, and the episomal stability of the pBH140 vector make this system ideal for studies on gene expression and other genomic functions in human cells. The potential significance of large, functionally active episomes for gene therapy is discussed. PMID- 9741426 TI - Inhibition of tumor necrosis factor alpha decreases inflammation and prolongs adenovirus gene expression in lung and liver. AB - The clinical application of adenoviral gene therapy currently is impeded by the potent host immune response to the virus, which limits the duration of its effects. In these studies, we investigated the role of TNF-alpha and of a soluble TNF receptor (TNF-bp) in the inflammatory response and expression of a lacZ expressing adenovirus (AdCMVlacZ) in the liver and lung of mice. The expression of the recombinant adenovirus was studied in mouse liver and lung by determining the activity of the lacZ gene product of the adenovirus. The mononuclear cell inflammatory response was determined histologically at different times after intravenous or intranasal administration of AdCMVlacZ. The cytotoxic T cell and antibody response to the adenovirus was determined. Treatment with TNF-bp reduced circulating levels of TNF-alpha, greatly reduced the inflammatory response, and resulted in prolonged expression of lacZ for up to 30 days in the liver and lung after either intravenous or intranasal administration of adenovirus. Treatment with TNF-bp had no effect on anti-adenovirus antibodies and induction of cytotoxic T cells 30 days after administration of AdCMVlacZ. These results indicate that TNF-alpha is the primary factor driving the early inflammatory response leading to elimination of adenovirus-infected cells in the liver and lung and that TNF-bp is capable of inhibiting these effects. PMID- 9741427 TI - Lung-specific expression of adenovirus E3-14.7K in transgenic mice attenuates adenoviral vector-mediated lung inflammation and enhances transgene expression. AB - Herein, we report that the adenovirus E3-14.7K protein inhibits the inflammatory response to adenovirus in transgenic mice in which the E3-14.7K gene was selectively expressed in the respiratory epithelium, using the human surfactant protein C (SP-C) promoter. E3-14.7K mRNA and protein were detected specifically in the lungs of SPC/E3-14.7K transgenic mice. Responses of the transgenic mice to Av1Luc1, an E1-E3-deleted Ad vector encoding the luciferase reporter gene, were examined, including vector transgene expression and lung inflammation. In wild type mice, luciferase activity declined rapidly and was lost 14 days following Av1Luc1 administration. The loss of luciferase activity was associated with pulmonary infiltration by macrophages and lymphocytes. In heterozygous SPC/E3 14.7K mice, luciferase activity was increased by 7 days compared with control littermates, and pulmonary infiltration by macrophages was decreased. In homozygous (+/+) SPC/E3-14.7K mice, luciferase activity was increased 7, 14, and 21 days following administration compared with wild-type mice, and lung inflammation was markedly reduced. After Av1Luc1 administration, PCNA staining of bronchiolar and alveolar respiratory epithelial cells was decreased in SPC/E3 14.7K transgenic mice, indicating decreased epithelial cell proliferation, a finding consistent with the observed reduction in inflammation. CD4 and CD8 lymphocyte populations were only mildly altered, while humoral responses to adenoviral vectors were unchanged in the SPC/E3-14.7K mice. The E3-14.7K protein expressed selectively in respiratory epithelial cells suppresses Ad-induced pulmonary epithelial cell cytotoxicity and lung inflammation in vivo and prolongs reporter gene expression. PMID- 9741428 TI - Genetic immunization with glycoprotein 63 cDNA results in a helper T cell type 1 immune response and protection in a murine model of leishmaniasis. AB - Genetic immunization is a promising gene therapy approach for the prevention and treatment of infectious disease. Plasmid DNA expressing genes of pathogens is directly introduced into host cells and specific cell-mediated and/or humoral immune responses are elicited against the encoded protein. Leishmaniasis is a significant world-wide health problem for which no vaccine exists. In susceptible animals, such as BALB/c mice, protection from leishmaniasis requires induction of a Thl immune response. In this study, cell-mediated immunity to Leishmania major (L. major) was induced by injecting BALB/c mice intradermally with plasmid DNA expressing the conserved L. major cell surface glycoprotein gp63 (gp63-pcDNA-3). CD4 T lymphocytes from gp63-pcDNA-3-immunized mice proliferated and produced IFN gamma (but not IL-4) when stimulated in vitro with freeze-thawed parasites, consistent with a Th1 immune response. In contrast, lymphocyte proliferation in animals immunized with freeze-thawed parasites was associated with IL-4 (but not IFN-gamma) production, suggesting a nonprotective Th2 response. Challenge studies revealed that gp63-pcDNA-3 vaccination protected 30% of susceptible mice (21 of 70) from Leishmania infection while neither gp63 protein (0 of 20) nor freeze thawed parasite vaccines (0 of 50) were efficacious. Dendritic cells derived from skin of gp63-pcDNA-3-injected mice also immunized naive recipients and protected them from leishmaniasis. We conclude that gp63-pcDNA-3 genetic vaccination results in a CD4-dependent Th1 immune response that correlates with protection from disease, and suggest that skin-derived dendritic cells are involved in priming this response. PMID- 9741429 TI - New helper cells and matched early region 1-deleted adenovirus vectors prevent generation of replication-competent adenoviruses. AB - The presence of replication-competent adenoviruses (RCAs) in batches of replication-defective adenovirus (Ad) vectors is a major problem for the application of these vectors in gene therapy. RCAs are generated by recombination between sequences in the Ad vector and homologous Ad sequences in the helper cells, resulting in the acquisition by the vector of early region 1. To prevent the formation of RCAs, we have developed helper cell lines, which we named PER, and matched Ad vectors that do not have sequence overlap. PER cells contain the Ad serotype 5 (Ad5) E1A- and E1B-encoding sequences (Ad5 nucleotides 459-3510) under the control of the human phosphoglycerate kinase (PGK) promoter. We demonstrate that PER cells synthesize high levels of the Ad5 E1A and E1B proteins. The yields from PER cells of E1-deleted Ads are similar to those obtained from earlier helper cells, such as 911 and 293 cells. Propagation of matched Ad vectors, which lack Ad5 nucleotides 459-3510, in one of the PER clones, PER.C6, does not result in the generation of RCAs, in contrast to propagation in 293 cells. We conclude that the combination of PER.C6 cells and nonoverlapping E1-deleted adenoviral vectors eliminates the problem of RCA generation by homologous recombination, and allows cost-effective production of safe, clinical-grade batches of recombinant Ad vectors. PMID- 9741430 TI - Heart-specific targeting of beta-galactosidase by the ventricle-specific cardiac myosin light chain 2 promoter using adenovirus vectors. AB - Adenoviruses are attractive vectors for gene transfer into cardiac muscle. However, their promiscuous tissue tropism, which leads to an ectopic expression of the transgene, is a considerable limitation. To restrict expression to cardiomyocytes, we have constructed two recombinant adenoviruses (Ad-MLC2 250betagal and Ad-MLC2-2100betagal) containing the beta-galactosidase reporter gene under the control of the 250- or 2100-bp rat ventricle-specific cardiac myosin light chain-2v promoter (MLC-2v). Our in vitro and in vivo data have evidenced that the 2100-bp promoter allows stronger beta-galactosidase activity than the 250-bp promoter and that the deleted promoter allows a weak beta galactosidase expression in skeletal muscle-derived cells in vitro. In contrast to the in vitro results, the highly deleted MLC-2v promoter of 250 pb conserved its heart specificity in in ovo and in vivo when introduced into the adenovirus genome, indicating that the specificity of this promoter is neither altered by the inverted terminal repeat nor by the enhancer of the Ela promoter, both of which located in the 5' flanking region of the promoter. Systemic injections of both recombinant adenoviruses into chicken embryos showed beta-galactosidase expression mainly in the right ventricle of the heart. We have confirmed the cardiac specificity of both promoters in mammalian species after injection of both recombinant adenoviruses into the heart of adult rats in vivo. The comparison of both promoters in vitro and in vivo has shown that the 250-bp MLC 2v promoter is 80% less active than the 2100-bp MLC-2v promoter and has enabled us to conclude that the MLC-2v promoter of 2100 bp is the most appropriate for efficient expression of a reporter gene or a therapeutic cardiac gene (e.g., SERCA2a or minidystrophin gene). PMID- 9741431 TI - Development of novel cell surface CD34-targeted recombinant adenoassociated virus vectors for gene therapy. AB - Recombinant adenoassociated virus (rAAV) type 2 vectors have been used to transduce a wide variety of cell types, including hematopoietic progenitor cells. For in vivo gene transfer, it is desirable to have an rAAV vector that specifically transduces selected target cells. As a first step toward generating an rAAV vector capable of targeting delivery in vivo, we have engineered a chimeric protein combining the AAV capsid protein and the variable region of a single-chain antibody against human CD34 molecules, a cell surface marker for hematopoietic stem/progenitor cells. Inclusion of the chimeric CD34 single-chain antibody-AAV capsid proteins within an rAAV virion significantly increased the preferential infectivity of rAAV for the CD34+ human myoleukemia cell line KG-1, which is normally refractory to rAAV transduction. Antibodies against the single chain antibody and the CD34 protein blocked this transduction. This chimeric vector represents a significant improvement in the host range of rAAV and the first step toward specific gene delivery by rAAV vectors to cells of choice, in this case, hematopoietic progenitor cells, for the treatment of human disease. PMID- 9741432 TI - Tetracycline repressor, tetR, rather than the tetR-mammalian cell transcription factor fusion derivatives, regulates inducible gene expression in mammalian cells. AB - This article describes the first (to our knowledge) tetracycline-inducible regulatory system that demonstrates that the tetracycline repressor (tetR) alone, rather than tetR-mammalian cell transcription factor fusion derivatives, can function as a potent trans-modulator to regulate gene expression in mammalian cells. With proper positioning of tetracycline operators downstream of the TATA element and of human epidermal growth factor (hEGF) as a reporter, we show that gene expression from the tetracycline operator-bearing hCMV major immediate-early enhancer-promoter (pcmvtetO) can be regulated by tetR over three orders of magnitude in response to tetracycline when (1) the reporter was cotransfected with tetR-expressing plasmid in transient expression assays, and (2) the reporter unit was stably integrated into the chromosome of a tetR-expressing cell line. This level of tetR-mediated inducible gene regulation is significantly higher than that of other repression-based mammalian cell transcription switch systems. In an in vivo porcine wound model, close to 60-fold tetR-mediated regulatory effects were detected and it was reversed when tetracycline was administered. Collectively, this study provides a direct implementation of this tetracycline inducible regulatory switch for controlling gene expression in vitro, in vivo, and in gene therapy. PMID- 9741433 TI - A phase I clinical trial of lethally irradiated allogeneic pancreatic tumor cells transfected with the GM-CSF gene for the treatment of pancreatic adenocarcinoma. PMID- 9741434 TI - The incidence of modular tibial polyethylene insert exchange in total knee arthroplasty when polyethylene failure occurs. AB - One of the primary reasons for utilizing modular tibial polyethylene inserts (MTPI) at the time of total knee arthroplasty is to have the ability to simply exchange the polyethylene at the time of revision surgery when polyethylene failure has occurred. During a 2-year period from January 1993 to December 1994, 62 revision total knee arthroplasties were reviewed from five different institutions in North America, which were performed secondary to modular tibial insert failure. In 55 cases (88.7%), significant scoring and/or damage to the femoral and/or tibial components occurred necessitating revision of one or both components. This series does not support the premise that polyethylene exchange is common at the time of revision surgery for MTPI failure. Of the patients 88.7% had MTPI failure resulted in femoral and/or tibial component revision secondary to surface damage to the femoral component and/or tibial component baseplate. In order to avoid femoral and tibial component wear secondary to MTPI failure, early recognition is necessary to allow the much simpler operation consisting of polyethylene liner exchange only. PMID- 9741435 TI - Effect of femoral head surface roughness on the wear of ultrahigh molecular weight polyethylene acetabular cups. AB - We studied the effect of femoral head surface roughness on the wear of ultrahigh molecular weight polyethylene (UHMWPE) acetabular cups using a hip joint simulator and a reciprocating wear tester. Compared with the hip simulator, the reciprocating wear tester severely exaggerates the effect of counterface roughness on UHMWPE wear and drastically underestimates the wear rate of the UHMWPE against smooth undamaged counterfaces. According to the hip simulator test results, the wear rate of the UHMWPE cups is approximately proportional to the square root of the femoral head roughness Ra (center-line-average roughness) rather than to Ra raised to a power greater than one as predicted by pin-on-disk studies. Roughening of the femoral heads by an order of magnitude results in a 2- to 3-fold increase in the wear rate. Therefore, the much wider clinical variations of wear cannot be fully explained by variations in surface roughness of the femoral heads. PMID- 9741436 TI - Effects of acetabular abduction on cup wear rates in total hip arthroplasty. AB - This study was designed to determine the effect of acetabular abduction on the polyethylene wear rates of the acetabular component. The hypothesis of this study is that acetabular placement, in particular abduction, effects contact forces and therefore polyethylene wear. A total of 364 total hip arthroplasties that were performed between 1974 and 1976 were included in this study. The cemented all polyethylene acetabular components were the same for each case and came from a single supplier. The polyethylene wear rates were calculated by measuring the shortest radius from the center of the prosthetic femoral head to a point on the outer surface of the acetabular cup. An immediate postoperative radiograph was compared with a follow-up radiograph at least 9.5 years later. A standardized radiograph was used to prevent differences due to magnification. The abduction or inclination of the acetabular cups was measured in all of the patients. The amount of acetabular cup abduction was measured relative to the ischial tuberosity line. The mean abduction was 44.1 degrees with a standard deviation of 9.2 degrees and a median of 44 degrees. The range of abduction was from 0 degrees to 85 degrees. This analysis failed to show a correlation between the amount of acetabular abduction and polyethylene wear rates (Pearson's correlation coefficient = 0.0679; P = .20). With a sample size of 364, there is over a 95% change (statistical power) of detecting an underlying true correlation between wear and abduction that is greater than or equal to 0.20. We were unable to demonstrate a difference in wear that would correlate with the differing degrees of abduction of the acetabular socket. We felt that the radiographic measurements of wear were quite accurate. This method of determining acetabular abduction has also been documented and supported in previous literature and has allowed us to accurately determine acetabular abduction. The results of our study demonstrate that within the normal ranges described by our study, polyethylene wear did not significantly increase with increased abduction of the acetabular component. PMID- 9741437 TI - Conversion of hemiarthroplasty to total hip arthroplasty: can groin pain be eliminated? AB - In this study 45 patients with groin or buttock pain after hemiarthroplasty were evaluated 2 to 7 years after conversion to total hip arthroplasty. Groin or buttock pain was completely relieved in 36 patients (80%) and partially relieved in an additional 4 patients (9%). After conversion surgery, 9 patients continued to have groin or buttock pain, but no factor could be identified that would predict an unsuccessful result. The hypothesis tested in this study was whether conversion of a hemiarthroplasty to a total hip arthroplasty eliminates groin pain. Because a significant number of patients (up to 20%) will continue to have some groin or buttock pain after conversion surgery, patients should be warned of this contingency before surgery. PMID- 9741438 TI - Bipolar shoulder arthroplasty for painful conditions of the shoulder. AB - Between April 1991 and March 1997, 182 bipolar shoulder replacements were implanted in 174 patients (8 bilateral) for painful conditions of the shoulder. The study group comprises 108 patients who were followed for an average of 2.9 years (range, 2-6 years). Diagnoses included osteoarthritis (51), rotator cuff arthropathy (27), avascular necrosis (3), revisions (8), rheumatoid arthritis (2), and fractures--both old and recent (17). A satisfactory rating (University of California at Los Angeles shoulder rating score greater than or equal to 28 points of 35) was achieved by 72% of the patients (including rotator cuff arthropathy patients). Patients with osteoarthritis obtained 90.2% of satisfactory results (46 of 51). The overall pain score after surgery was 8.8 points (of 10), meaning that none or occasional pain was present. Five patients required prosthetic revision, and 102 (94.4%) were satisfied with the surgical procedure. PMID- 9741439 TI - Migration and failure of the Mecron screw-in acetabular prosthesis. AB - Early results with the Mecron prosthesis have been variable. We report our experience with this prosthesis at medium- to long-term follow-up. At 5 to 9 years after surgery, 43 patients (49 implants) were reviewed. Radiographic measurements of superior migration, alteration in the opening angle, and thread engagement were made. The failure rate was very high, 33% having been revised or awaiting revision. Of the remainder, 80% had migrated, 86% had changes in the opening angle, and 65% had fewer threads engaged. Many have low-grade symptoms insufficient to merit revision, but the extent of loosening makes eventual revision likely. We feel this represents an example of a new design gaining widespread usage before adequate follow-up studies. PMID- 9741440 TI - Rapidly progressive osteoarthrosis of the hip. AB - Rapidly progressive cases of primary idiopathic hip osteoarthrosis are well known and recognized. Their prevalence and significance are, however, often poorly appreciated. This article aims to assess the prevalence of the condition in a United Kingdom district general hospital. Several points arise: 1) Patients suffering rapid deterioration do not always alert their surgeon to the worsening symptoms; 2) Waiting lists need regular administration and the patients regular review while on such lists; 3) Decisions about surgery and case selection should be made purely on clinical grounds with financial, political, and administrative considerations accorded little or no relevance. PMID- 9741441 TI - Hip arthroplasty in chronic dialysis patients. AB - We retrospectively reviewed 15 patients (24 hips) on chronic renal dialysis who underwent hip arthroplasty between 1970 and 1990. The average age at surgery was 39 years; the average follow-up was 8 years (range, 1-19 years). All follow-up of less than 5 years relates to those patients who died. Of these 24 hips, 14 (58%) failed or were failing due to loosening; the average time to revision was 7 years (range, 1.5-14 years). A complicated course was experienced in 16 hips (66%), primarily related to medical difficulties. There was one perioperative death. The following orthopedic complications afflicted 5 hips (21%): one femur fracture during revision; one femur fracture at 2 months after revision; one dislocation during seizure; one displacement of acetabular cup requiring recementing; and late generalized septic death of one patient (with both hips involved). Within an average of 3 years (range, 15 months to 7 years) after their index surgery, 6 of the 15 patients (40%) died. The patients who lived were chronically ill, and all but three remained on long-term dialysis. The functional level of all those remaining on dialysis steadily declined, and none reached a quality of life comparable to an osteoarthritic patient. This study confirms a previously reported high mortality and morbidity rate in this population. Despite their difficulties, 22/24 primary hips were relieved of pain and increased in function; six patients returned to work. We see no better alternative for pain relief in total hip arthroplasty, particularly in view of contemporary surgical techniques and improved medical management. PMID- 9741442 TI - Intraoperative type 1 proximal femoral fractures: influence on the stability of hydroxyapatite-coated femoral components. AB - We reviewed a series of 120 uncemented total hip replacements using the Omniflex stem with hydroxyapatite coating. Twenty minor intraoperative proximal fractures occurred. All fractures were treated with cerclage wiring after removal of the stem. Radiographic and clinical results of these 20 patients were compared with the remaining 100 implants in which this complication did not occur. In 20% of the cases of both groups, a migration of less than 2 mm was observed. No differences were detected in Harris Hip Scores, subsidence of the stem, and radiographic behavior. We concluded that a properly stabilized proximal femoral fracture above the lesser trochanter did not influence the clinical and radiographic results at more than 3 years follow-up. PMID- 9741443 TI - Rheumatoid arthritis and hydroxyapatite-coated hip prostheses: five-year results. International ABG Study Group. AB - Medium- and long-term results from cemented total hip arthroplasty (THA) in patients with rheumatoid arthritis (RA) show a higher incidence of infection and aseptic loosening when compared to other diagnoses. Early results using uncemented prostheses are variable. Hydroxyapatite (HA) coatings are thought to improve early osseointegration in uncemented THA. In a prospective, international, multicenter clinical study, 32 RA patients who received 33 uncemented HA-coated prostheses were followed up for a minimum of 5 years. Medium term clinical results are excellent. No infection or aseptic loosening has been recorded. Excellent osseointegration was observed radiographically. Bone remodeling was consistent with mainly proximal to midstem stress transfer. We conclude that uncemented, proximally HA-coated prostheses are a promising alternative to cemented prostheses for RA patients. PMID- 9741444 TI - Wear of bipolar hip prostheses. AB - A method was developed to take radiographs showing the inner articulation of bipolar hip prostheses. By this method, wear was measured in 68 hips whose inner head diameter was 22 mm. Average annual wear rate was 0.17 mm. Osteolysis was observed in 25 hips (37%) and there was no difference between the annual wear rate of hips with and without osteolysis. Studying 19 retrieved prostheses, abrasion of the rim was deeper in hips with osteolysis than those without it. Wear rate of the inner articulation in bipolar hip prosthesis is much larger than that in Charnley's prosthesis, as linear penetration into the articulation surface reduces the motion range of the inner articulation and this increases impingement and advances rim abrasion. PMID- 9741445 TI - Reconstruction of the deficient acetabulum with an oblong prosthesis: three- to seven-year results. AB - A retrospective review was conducted to determine the intermediate-term results of a noncustomized, oblong, porous-coated, cementless, acetabular component designed to obtain stability on host bone while maintaining an anatomic hip center. The clinical and radiographic results were reviewed in 18 patients (3 primary, 15 revision). All patients had substantial acetabular rim defects before reconstruction with the Johnson and Johnson E-15 or E-25 oblong components. The average follow-up was 4.5 years (range, 3.4-6.9 years), with an average postoperative Harris Hip Score of 91 points. Radiographic analysis revealed no prosthetic migration and near anatomic restoration of the hip center. PMID- 9741446 TI - Revision total knee arthroplasty using the porous-coated anatomic revision prosthesis: six- to twelve-year results. AB - The porous-coated anatomic (PCA) revision prosthesis was one of the earliest revision total knee systems to feature more constraint for stability, stems for fixation, and thicker femoral components to address bone loss. From 1981 to 1989, 36 revision total knee arthroplasties were performed using the PCA prosthesis. Patients were evaluated by clinical examination, radiographs, and the Hospital for Special Surgery Knee Rating Scale. Six patients died and three were lost to follow-up; 27 knees remained for follow-up. The average age at surgery was 66 years. Follow-up averaged 9.8 years (range, 6-12.4 years). Range of motion averaged 4 degrees to 91 degrees before surgery and 1 degrees to 92 degrees after surgery. Knee scores averaged 48 before surgery and 86 after surgery. Three femoral and 10 tibial components showed lucent lines, with 3 tibial components showing progressive lucency. Six patients required reoperation: 3 for tibial component loosening, 1 for wound infection, 1 for deep infection, and 1 for persistent pain. These results suggest that the use of an implant specifically designed for revision can yield successful long-term results. PMID- 9741447 TI - Increased urinary crosslink levels in aseptic loosening of total hip arthroplasty. AB - The diagnosis of aseptic loosening in total hip arthroplasty is predominantly based on clinical and radiographic evaluation. Loosening is usually associated with increased bone resorption at the interface. In this study we wanted to evaluate the diagnostic value of bone markers in aseptic loosening. We compared 50 patients with proven component loosening during surgery with 50 age-, sex-, and implant-matched patients without clinical or radiological signs of loosening. We measured serum markers of bone formation (bone-specific alkaline phosphatase, osteocalcin [OC], procollagen type I propeptides) and bone resorption (collagen n telopeptide [NTX], deoxypyridinoline [DPYD], pyridinoline [PYD]). We found significantly increased levels of NTX, DPYD, PYD, and OC in the loosening group. The other markers showed no significant difference between both groups. We conclude that determination of urinary crosslinks may offer a new and valuable diagnostic method in the detection of aseptic loosening in total hip arthroplasty. PMID- 9741448 TI - Contact areas and pressures between native patellas and prosthetic femoral components. AB - Contact areas and pressures between native patellas and a prosthetic condylar design femoral component were measured at flexion angles of 30 degrees, 60 degrees, and 90 degrees. These were compared to measurements obtained with a domed all-polyethylene patellar component. Mean native patellar contact areas were found to be fourfold greater than seen with the prosthetic patellar component. Contact stresses in the native patellas were below the yield strength of articular cartilage in 80% of the contact area. By contrast, stresses measured in the prosthetic patella exceeded the yield strength of ultrahigh molecular weight polyethylene in 64% of the measured contact area. Contact areas and stresses were not significantly effected by flexion angle. Although contact areas and stresses reflect only a part of the dynamics of the patellofemoral articulation this information would support the selective retention of the native patella in total knee arthroplasty. PMID- 9741449 TI - Contact stress analysis in meniscal bearing total knee arthroplasty. AB - The effect of a mobile meniscal bearing on tibiofemoral contact stress was tested with a standard fixed tibial component and with movable tibial components (anteroposterior sliding, rotationally sliding, and anteroposterior and rotationally sliding). A digital electronic sensor was used to detect tibiofemoral contact location in five cadaver knees, then the location was reproduced while peak and mean stresses were measured under compressive load at 0 degrees, 30 degrees, 60 degrees, and 90 degrees of flexion. Stresses were measured when the tibial component was normally aligned and at 15 degrees internal and 15 degrees external rotation. To evaluate the effect of excessive overhang of the polyethylene articular surface, undersurface stress of the rotationally sliding component was also measured with a 30 degrees and a 45 degrees malrotated tibial tray. Uppersurface stresses of the fixed-bearing components were significantly higher at full extension than those recorded in components with rotational mobility. Undersurface stresses were always lower than uppersurface stresses, but correlated with uppersurface stresses. Undersurface stresses of the rotationally sliding component gradually increased as the malrotation angle of the tray increased. A mobile meniscal bearing surface appears to offer an advantage over a standard fixed component when rotational malalignment of the tibial component occurs. However, with severe rotational malalignment, edge contact markedly increases undersurface stresses, which could cause deformity and subluxation. PMID- 9741450 TI - Successful salvage of a primary total knee arthroplasty infected with Candida parapsilosis. AB - Fungal infections of total joint arthroplasties are extremely rare with only 21 previous reported cases in the literature. In 19 of these cases, the offending organism has been a member of the candida species. In all of these cases, the patients had no clinical evidence of disseminated fungal infection. All previously reported cases of total joint fungal infections required removal of the primary prosthesis to eradicate the infection. There has also been a great reluctance to reimplant these patients. In fact, reimplantation has been successful in only one reported case. We report the first case of successful salvage of an arthroplasty infected with candida. PMID- 9741451 TI - Drain entrapment and titanium to ceramic head deposition: two unique complications following closed reduction of a dislocated total hip arthroplasty. AB - Postoperative dislocation remains one of the most frequent complications following total hip replacement. In this report, a case is presented that illustrates two potential concerns with postoperative dislocation and subsequent closed reduction. The first complication presented in this report is entrapment of a closed drainage system tube in the joint space following closed reduction. The second complication, transfer of metallic debris to a ceramic femoral head from contact with an acetabular shell during closed reduction, was documented by analysis of a femoral head using scanning electron microscopy and energy dispersive x-ray spectrometry. This report emphasizes the need for the surgeon to express caution when relocating a dislocated hip, particularly when a closed drainage system is used postoperatively. PMID- 9741452 TI - Prophylactic intramedullary femoral rodding during total knee arthroplasty with simultaneous femoral plate removal. AB - Removal of a plate from the distal femur creates a risk of fracture through the screw holes. This is a particular concern when a total knee arthroplasty is present because supracondylar fracture may occur with minimal trauma. A patient who presents after prior plating of a distal femur fracture with osteoporosis, retained hardware associated with pain, and gonarthrosis severe enough to warrant total knee arthroplasty is often difficult to manage. Prophylactic intramedullary rodding is a well-accepted method of treating pathologic stress risers in the femur. An intramedullary rod can be inserted into the femur at the time of total knee arthroplasty. This method permits simultaneous plate removal and total knee arthroplasty while protecting the femur from postoperative fracture. PMID- 9741453 TI - Effect of patellar meniscus on patellofemoral contact stress in total knee arthroplasty. AB - The effect of a meniscus of fibrous tissue (patellar meniscus), on patellofemoral contact area and stresses was evaluated. Two knees with total knee arthroplasty and dome-shaped patellar components were retrieved at autopsy. Both had substantial fibrous menisci surrounding the dome of the patellar component. Contact area and contact stresses were measured with a digital electronic sensor, first with the patellar meniscus intact, then again after the patellar meniscus was removed. No dramatic difference was detected in patellofemoral contact area and contact stresses between the patellas with an intact or removed patellar meniscus, and there was no detectable load under the patellar meniscus. The results of this case study suggest that fibrous tissue surrounding the dome shaped patellar component does not share compressive loads with the patellar component. PMID- 9741454 TI - Acute vascular rejection. PMID- 9741455 TI - Extracorporeal ("ex vivo") connection of pig kidneys to humans. III. Studies of plasma complement activation and complement deposition in the kidney tissue. AB - The complement system is one of the important factors involved in the hyperacute rejection of xenografts. This report deals with the activation of the complement system in a clinical trial where pig kidneys were extracorporeally connected to two volunteer dialysis patients who were pretreated with plasmapheresis in order to substantially reduce anti-pig xenoantibodies. The clinical data of the perfusion experiments and the patients humoral immune response to pig xenoantigens have been reported in detail (Xenotransplantation 1996; 3:328-339, 340-353). Three consecutive daily plasmapheresis treatments of the patients reduced the plasma complement protein (C3, C4, and C5) concentrations to 8-27% of the baseline values. The perfusion of the pig kidney connected to patient 1 was terminated at 65 min due to graft rejection and this patient was not hemodynamically affected by the experiment. The second experiment was terminated at 15 min due to an anaphylactic like reaction of the patient. In patient 1 a slight reduction of plasma C3, C4, and C5 and an increase of C5a and SC5b-9 occurred, while C3a decreased during the perfusion. Patient 2 had an increase of all complement parameters, most prominent for C4d and SC5b-9, which occurred concomitant with the appearance of the anaphylactic like side effects. In general, plasma levels of PMN elastase, IL6 and IL8 increased in both patients during the perfusion. Immunohistochemical investigation of the kidney tissues revealed deposition of human complement factors C1q, C4c, and C3c in a congruent pattern with the vasculature of the kidney in patient 1. In kidney 2 only trace amounts of C1q and C3c were found. Both kidneys were negative for properdin. Therefore, in this experimental set up with extracorporeal connection of pig kidneys to the human circulation the human complement cascade is activated mainly through the classical pathway. PMID- 9741457 TI - Adult and neonatal anti-Gal response in knock-out mice for alpha1,3galactosyltransferase. AB - The knockout mouse to alpha1,3galactosyltransferase (alpha1,3GT KO) lacks the ability to synthesize alpha-gal epitopes (Galalpha1,3Galbeta1,4GlcNAc-R) and is capable of producing low amounts of the natural anti-Gal antibody. The present study indicates that repeated immunization of these mice with rabbit red blood cell (RRBC) membranes results in production of anti-Gal in titers and specificity similar to those in humans. In contrast, immunized wild-type mice completely lack anti-Gal. Anti-Gal in the alpha1,3GT KO mice is produced in the circulation as the various IgG subclasses and as IgM isotype, but not IgA. In view of previous reports on the possible induction of T cell tolerance by immunization of mice with large amounts of antigen up to 24 days of age, we assayed possible induction of neonatal B cell tolerance toward the alpha-gal epitope. Eight-day-old neonates were subjected to immunization with 1 x 10(8) RRBC membranes, or 30 x 10(6) wild type mouse splenocytes, both of which express an abundance of alpha-gal epitopes. These neonatal exposures to alpha-gal epitopes did not prevent subsequent production of anti-Gal. Thus, the tolerance induction to this carbohydrate epitope is likely to be mediated by mechanisms other than those inducing neonatal T cell tolerance. PMID- 9741456 TI - High-level co-expression of complement regulators on vascular endothelium in transgenic mice: CD55 and CD59 provide greater protection from human complement mediated injury than CD59 alone. AB - High-level endothelial expression of the human complement regulatory factor CD59 has been shown to protect transgenic mouse hearts from human complement-mediated injury in an ex vivo perfusion model. In this study we examine whether co expression of CD55 provides additional protection. CD55/CD59 double-transgenic mice were generated by co-injection of CD55 and CD59 expression constructs driven by the human intercellular adhesion molecule 2 (ICAM-2) promoter. A line was established from one mouse that exhibited strong expression of CD55 and CD59 on vascular endothelium in the heart and other transplantable organs. An ex vivo perfusion model was used to compare hearts from these CD55/CD59 mice with hearts from a previously established line, which expressed CD59 at a similar level to the double transgenic line. CD59 hearts displayed prolonged survival compared to wild-type hearts during perfusion with 40% human plasma and maintained approximately 20% maximum work after 60 min. CD55/CD59 hearts were further protected, with work maintained at 35% of the maximum level after 60 min. The data demonstrate that high-level endothelial co-expression of CD55 and CD59 provides greater protection from human complement-mediated injury in this model than expression of CD59 alone. PMID- 9741458 TI - Comparative histopathology of hepatic allografts and xenografts in the nonhuman primate. AB - Liver transplantation was performed in the following groups: Group 1, baboon-to baboon allografting (n=8) (control group); Group 2, ABO-compatible vervet monkey to-baboon xenografting (n=8); Group 3, ABO-incompatible vervet monkey-to-baboon xenografting (n=6); Group 4, pig-to-baboon xenografting (n=2); and Group 5, pig to-rhesus monkey xenografting (n=6). Immunosuppressive therapy (cyclosporine, cyclophosphamide, and methylprednisolone) was begun 2-7 days before liver transplantation (LTx) and continued indefinitely after LTx. The liver grafts were biopsied pre-LTx and subsequently post-LTx at approximately 1 hr, 2-3 hr, 7-10 days, 20-30 days, 60 days, 120 days, and at euthanasia or spontaneous death. There were 19 successful LTxs with grafts functioning from one hour to 123 days. No pig liver (Groups 4 and 5) survived more than 5.5 hr, as there was an immediate severe vascular response after reperfusion, typical of hyperacute rejection (congestion and hemorrhage). Vascular rejection was not seen in allografts (Group 1), but early mild-to-moderate congestion and neutrophil infiltration were present in concordant xenografts (Groups 2 and 3), which were associated with moderate deposition of immunoglobulin, C3, and fibrinogen. Lymphoid cell infiltration, bile duct damage, and portal vein endothelialitis in the portal zones occurred later in both allografts (Group 1) and concordant xenografts (Groups 2 and 3), developing earlier in the presence of ABO incompatibility (Group 3). In concordant xenografts it was usually followed by fibrosis. PMID- 9741459 TI - Requirement of CD4 cells for induction and maintenance of unresponsiveness in islet xenografted mice. AB - Long-term survival of islet xenografts in the hamster to mouse model can be induced by a short-course treatment with a nondepleting anti-CD4 mAb but not with a depleting anti-CD4 mAb [Lu et al. Xenotransplantation 1998; 5:154-163]. Although CD4 cells are known to play a key role in the rejection of islet xenografts, it remains unclear whether CD4 cells are also required for the induction and/or maintenance of specific unresponsiveness to xenografts. To investigate this problem, islets were isolated from golden hamsters and transplanted into streptozotocin-induced diabetic CBA/J mice. Nondepleting mAb YTS 177.9 was used to block CD4 cells for the induction of islet xenograft unresponsiveness and subsequently depleting mAb GK1.5 to deplete CD4 cells in the unresponsive recipients. First, we now confirm that second donor-strain xenografts were permanently accepted in recipients that had been unresponsive to the first grafts, whereas Lewis rat islet xenografts, used as third-party grafts, were rejected like a primary graft within 7-8 days. Second, we depleted CD4 cells in recipient mice, which had been treated perioperatively with the nondepleting mAb YTS 177.9 and became unresponsive to their primary hamster islet graft, by using a depleting anti-CD4 mAb at different time points post-transplant. Depletion of CD4 cells in the unresponsive recipients by the depleting anti-CD4 mAb GK1.5 did abrogate this unresponsive state, since the grafts were always rejected within an average of 25.5 days after the mAb GK1.5 injections. Therefore, our results strongly suggest that CD4 positive cells play an active suppressive role and that their presence in the recipients appears essential for both induction and maintenance of long-term islet xenograft survival or specific unresponsiveness. PMID- 9741461 TI - Survival of fetal porcine pancreatic islet tissue transplanted to a diabetic patient: findings by ultrastructural immunocytochemistry. AB - Porcine fetal endocrine pancreatic tissue was placed under the kidney capsule in a diabetic renal transplant patient. In a core-needle kidney biopsy specimen obtained 3 weeks after transplantation, clusters of epithelial cells were identified in the subcapsular space. The ultrastructural and immunocytochemical features of these cells were typical of pancreatic islet cells. Some cells stained positively for insulin; others stained positively for glucagon, somatostatin or chromagranin A. There were well-defined cytoplastic storage and transport granulae that indicated hormone synthesis. The ultrastructural findings provide further evidence that porcine cells can survive after transplantation to humans. PMID- 9741460 TI - Cyclophosphamide, but not CTLA4Ig, prolongs survival of fetal pig islet grafts in anti-T cell monoclonal antibody-treated NOD mice. AB - Fetal pig islets, xenografted after organ culture into non-immunosuppressed prediabetic NOD mice, are rejected within 10 days. Immunosuppression with anti-T cell (anti-CD4 and anti-CD3) monoclonal antibodies alone is highly effective in delaying graft rejection in this discordant model, but rejection eventually occurs, usually within 80 days, despite marked depletion of T cells. In an attempt to prevent rejection, we used cyclophosphamide (CP), a powerful anti-B cell agent, or CTLA4Ig, an inhibitor of T-cell co-stimulation [via B7-1 (CD80) and B7-2 (CD86)], either given in combination with anti-CD4 (GK1.5) or anti-CD3 (KT3) MAb to the recipient mice. The addition of cyclophosphamide in a dose that significantly depleted B cells in peripheral blood was highly effective in preventing rejection, with xenografts surviving for at least 112 days, when the experiment was terminated. CTLA4Ig, administered alone, did not prevent delayed rejection (rejection occurred in <60 days) and, in contrast to CP, did not prevent delayed rejection when used in combination with GK1.5 and KT3 treatment. Thus, immunosuppressive agents found to be highly effective in other strains, e.g., CTLA4Ig and anti-T cell MAbs, had a lesser effect in NOD mice but the addition of an anti-B cell drug, CP, was useful. This finding may be applicable to patients with IDDM. PMID- 9741462 TI - Literature update 1998, Part 1. PMID- 9741463 TI - Permanent Committee of the International Congress of Human Genetics. AB - For over three decades, the Permanent Committee of the International Congresses of Human Genetics has served the purpose of selecting a host and site for the quadrennial Congress, which is due next in 2001. The Committee has statutes and consists of one voting representative from 38 nations and other ex officio members, much like the General Assembly of the United Nations. It meets twice during each International Congress, otherwise conducting its business by mail and telecommunication. Its structure could be the beginnings of a truly global democratic body of human geneticists to address other issues and to serve other purposes, as human genetics becomes increasingly international, as inevitably it must. PMID- 9741464 TI - Interstitial duplications of chromosome region 15q11q13: clinical and molecular characterization. AB - Duplications of chromosome region 15q11q13 often occur as a supernumerary chromosome 15. Less frequently they occur as interstitial duplications [dup(15)]. We describe the clinical and molecular characteristics of three patients with de novo dup(15). The patients, two males and one female (ages 3-21 years), had nonspecific findings that included autistic behavior, hypotonia, and variable degrees of mental retardation. The extent, orientation, and parental origin of the duplications were assessed by fluorescent in situ hybridization, microsatellite analyses, and methylation status at D15S63. Two patients had large direct duplications of 15q11q13 [dir dup(15)(q11q13)] that extended through the entire Angelman syndrome/Prader-Willi syndrome (AS/PWS) chromosomal region. Their proximal and distal breaks, at D15S541 or D15S9 and between D15S12 and D15S24, respectively, were comparable to those found in the common AS/PWS deletions. This suggests that duplications and deletions may be the reciprocal product of an unequal recombination event. These two duplications were maternally derived, but the origin of the chromatids involved in the unequal crossing over in meiosis differs. In one patient, the duplication originated from two different maternal chromosomes, while in the other patient it arose from the same maternal chromosome. The third patient had a much smaller duplication that involved only D15S11 and parental origin could not be determined. There was no obvious correlation between phenotype and extent of the duplication in these patients. PMID- 9741465 TI - Genetic epidemiology study of idiopathic talipes equinovarus. AB - Previous genetic studies of idiopathic talipes equinovarus (ITEV) suggest an environmental and genetic component to the etiology of ITEV. The present study was undertaken to assess the role of causal factors in the development of ITEV. A total of 285 propositi were ascertained, with detailed family history information available in 173 cases and medical records on the remaining 112 propositi. Information was collected on specific prenatal, parental, and demographic factors. No racial heterogeneity was noted among any of the factors. The overall ratio of affected males to females was 2.5:1. The incidence of twinning among all propositi was significantly increased (P=0.006) above the expected population frequency. A family history of ITEV was noted in 24.4% of all propositi studied. These findings, in addition to the detailed analysis of 53 pedigrees with ITEV history, suggest the potential role of a gene or genes operating in high-risk families to produce this foot deformity. PMID- 9741466 TI - Segregation analysis of idiopathic talipes equinovarus in a Texan population. AB - "Idiopathic" talipes equinovarus (ITEV) is a nonsyndromal congenital anomaly of one or both feet. Casting and surgery are often necessary to obtain correct foot alignment. In spite of treatment, residual deformities of the feet occur and calf muscles may be hypoplastic. The cause of ITEV is unknown but genetic factors have been postulated. Complex segregation analysis was performed on 173 ITEV families including 93 Caucasian and 48 Hispanic families. The recessive mixed model was the best fitting model and no differences were found based on ethnicity. These results confirm previous studies suggesting that there is a genetic component to the development of ITEV. PMID- 9741467 TI - Partial tetrasomy with triplication of chromosome (5) (p14-p15.33) in a patient with severe multiple congenital anomalies. AB - We report on a newborn infant with a de novo triplication of the distal segment of 5p: 46,XX,trp(5) (pter-->p14::p14-->p15.33::p15.33--> qter) and multiple congenital anomalies consistent with triplication of 5p. Partial triplication was documented by fluorescence in situ hybridization with a cosmid probe specific for 5p15.2 and microdissected probes obtained from "5pter." Partial duplication of the short arm of chromosome 5 is associated with a specific phenotype that appears to be dependent on the chromosomal region duplicated. Duplication of 5p with breakpoints proximal to band p14 is generally associated with distinct craniofacial malformations, cardiac, renal, intestinal, and limb defects, and mental retardation, whereas duplications with breakpoints distal to 5p14 result in a milder phenotype characterized by minor facial anomalies, developmental delay, and seizures. The most proximal breakpoints of the partial triplication in this patient was estimated to be 5p14, suggesting that a more severe phenotype can occur with triplication of the more distal segment. PMID- 9741468 TI - Survival of children with Down syndrome in South America. ECLAMC-Downsurv Group. Latin American Collaborative Study of Congenital Malformations. AB - The first step of all healthcare actions aimed at promoting an appropriate quality of life for infants affected by Down syndrome (DS) is to ensure their survival. This investigation was aimed at estimating the infant mortality rate of infants affected with DS in urban populations of South America. Thirty-three hospitals included in the Latin American Collaborative Study of Congenital Malformations (ECLAMC) distributed in 23 cities of 5 South American countries followed 360 liveborn DS cases born during the 1988-1992 period. Families were recontacted after the infant should have reached the age of one year. The collected data included information about health status; i.e., frequency and dates of diagnosed illnesses and hospital admissions, and, in case of death, information on date, place and cause of death, and illness immediately before death. Information about the interviews included place, date, and name of the interviewer. A closed questionnaire was employed by the interviewers, mostly physicians, nurses, and social workers. Life table analysis up to the age of one year was performed by the actuarial survival method. The overall mean survival at age one year was 0.736 (SE=0.023). Thirty-three (9.2%) of the 360 cases died neonatally, and 62 (17.2%) within the remaining 2-to-12-month interval. The probability of survival at one year of age did not differ between public (209 cases; mean 0.718; SE=0.031) and private (151 cases; mean: 0.762; SE=0.035) (chi2:0.87; df:1; P >0.05) health systems. The 150 DS cases with a congenital heart defect (CHD) had a significantly lower P robability of survival at the age of one year (mean: 0.660; SE: 0.039) than did the 210 cases without CHD (mean: 0.790; SE: 0.028) (chi2:6.67; df:1; P <0.01). The death rate in the first year of life for DS cases without a detected cardiac defect (21%) is significantly higher than that reported in developed countries; namely, 16% from Italy, 11% from Canada, 10% from England, and 7% from Denmark. PMID- 9741469 TI - Thirteen cases of Niikawa-Kuroki syndrome: report and review with emphasis on medical complications and preventive management. AB - Eight new and five previously illustrated patients with Niikawa-Kuroki syndrome (NKS) are compared to those in the literature, providing data on 183 cases. Eight patients had disproportionate microcephaly and in one autopsied patient there was frontal lobe atrophy, focal polymicrogyria, and a hypoplastic fourth ventricle. The metacarpophalangeal pattern profiles of three Caucasian patients with NKS were similar to that of a prior case report, but those of two Hispanic patients were more variable. NKS was eliminated by follow-up in nine suspect cases, highlighting the diagnostic value of findings such as arched eyebrows, long palpebral fissures, flat nasal tip, and prominent finger pads. One patient suspected of having NKS had a very different metacarpophalangeal pattern profile, supporting its diagnostic utility in selected cases. Higher frequencies of neonatal complications, abnormal dentition, hypotonia, and microcephaly were noted in non-Asian patients with NKS, while a higher frequency of skeletal anomalies was seen in Japanese patients. Complications affecting cognitive, visual, hearing, cardiac, renal, skeletal, immune, and endocrinologic functions are translated into a program for preventive management. X chromosome anomalies are the most compelling of diverse genetic changes seen in NKS, and this report adds another case to several possible instances of vertical transmission. The 108 non-Asian patients now reported emphasize the worldwide significance of NKS recognition. PMID- 9741470 TI - Trisomy 16 and trisomy 16 Mosaicism: a review. AB - A review of all prenatal and postnatal diagnoses of trisomy 16 and trisomy 16 mosaicism was carried out in the context of the current understanding of confined placental mosaicism and uniparental disomy (UPD). The prenatal detection of trisomy 16 cells is associated with a high probability of fetal death, preterm delivery, intrauterine growth retardation, and fetal anomalies. Birth defects were typical of those seen in nonmosaic partial duplications of chromosome 16. Surprisingly, anomalies were sometimes limited to a single organ and included some relatively common isolated defects such as a ventricular septal defect, hypospadias, imperforate anus, inguinal hernia, and clubfoot. The risk for abnormality appeared to be higher in those pregnancies in which trisomy 16 cells were identified in amniotic fluid compared to the detection in chorionic villi samples. Contrary to nonmosaic trisomy 16 with an excess of males, mosaic trisomy 16 shows an excess of female karyotypes. Following the prenatal detection of trisomy 16 cells, aneuploid cells are almost never found in fetal or neonatal lymphocytes. Studies on fibroblasts also often fail to confirm the presence of the abnormal cell line even in cases in which multiple anomalies are present. It is likely that trisomy 16 cells are sometimes present in the early developing embryo even though subsequent cytogenetic studies on fetal or neonatal tissues may not detect any aneuploid cells. UPD can be excluded as a mechanism for those anomalies that are common to mosaic trisomy 16 and nonmosaic partial duplications. The term "occult mosaicism" is suggested to describe the situation in which the presence of an abnormal cell line is suspected on the basis of clinical data but unproven by laboratory analysis. PMID- 9741471 TI - Sublocalization of the Papillon-Lefevre syndrome locus on 11q14-q21. AB - Papillon-Lefevre syndrome (PLS) is an autosomal recessive form of palmoplantar ectodermal dysplasia, characterized by palmoplantar hyperkeratosis and severe early-onset periodontitis. The presence of severe periodontitis distinguishes PLS from other palmoplantar keratodermas. As part of our efforts to study the genetic basis of periodontitis susceptibility, we performed a genome-wide search to identify major loci for PLS in 44 individuals (14 affected) from 10 consanguineous PLS families. We have identified evidence for linkage of a PLS gene on 11q14-q21. A maximum two-point logarithm of the odds (LOD) score of 8.24 was obtained for D11S1367 at a recombination fraction of theta=0.00. Multipoint analysis resulted in a LOD score of 10.45 and placed the gene for PLS within a 4 5 cM genetic interval. This genetic interval, flanked by D11S4197 and D11S931, contains more than 50 cDNAs and 200 expressed sequence tags (ESTs). This refinement of the candidate region for a PLS gene is in agreement with other recent reports of linkage for PLS to chromosome 11q14-q21 and should help in identification of the gene for PLS. PMID- 9741472 TI - Transition to young adulthood in Ullrich-Turner syndrome: neurodevelopmental changes. AB - Studies describing the neurocognitive profile of Ullrich-Turner syndrome (UTS) have focused primarily on neurodevelopmental changes in childhood and adolescence or in adults with UTS. The objective of the present study was to describe neurodevelopmental changes that occur in UTS females during the transition from adolescence to young-adulthood. The subjects included 99 females with UTS and 89 normal female controls matched for age and socioeconomic status. Subjects were between the ages of 13 and 21 years. All subjects received a battery of neurocognitive tests designed to assess general cognitive ability, academic achievement, memory, language, executive function, visual-spatial/perceptual and motor skills, affect recognition, attention, and motor skills. Results from our study indicated that females with UTS performed significantly less well than controls on measures of spatial/perceptual skills, visual-motor integration, affect recognition, visual memory, attentional abilities, and executive function, consistent with previous reports of cognitive abilities in adolescent UTS females. Moreover, our results indicate that decreased performance in some of these areas persists through late adolescence and into early adulthood while improvement occurs in other areas. It is possible that catch-up in certain cognitive deficiencies in UTS females represents a maturational/developmental lag. Alternatively, the neurodevelopmental changes that were observed in UTS females may result from the cumulative effects of estrogen replacement therapy during adolescence. Therapeutic interventions specific to the demands of young adulthood are also discussed. PMID- 9741473 TI - Familial migraine with vertigo: no mutations found in CACNA1A. AB - We searched for mutations in the voltage-gated calcium channel gene, CACNA1A, in nine propositi of families with migraine headaches and episodic vertigo inherited in an autosomal dominant pattern. All 47 exons and flanking introns in CACNA1A were subjected to single-strand conformation polymorphism analysis of polymerase chain reaction-amplified genomic DNA. Exons with aberrantly migrating fragments were sequenced using standard techniques. We also determined the CAG repeat length at the 3' end of CACNA1A. Several polymorphisms were found but no mutations identified in any of the 47 exons of the 9 patients. No index-case had a CAG repeat length greater than 13 (normal <17). Mutations in CACNA1A are not common in families with migraine headaches and episodic vertigo. Other ion channel genes expressed in the brain and inner ear remain candidate genes. PMID- 9741474 TI - Anterior laryngeal webs and 22q11 deletions. PMID- 9741475 TI - Noonan syndrome: genotype analysis of the Noonan syndrome critical region at chromosome 12q in a three-generation family. PMID- 9741476 TI - Peripheral target-specific influences on embryonic neurite growth vigor and patterns. AB - We examined axon-target interactions in cocultures of embryonic rat trigeminal, dorsal root, nodose, superior cervical ganglia or retina with a variety of native or foreign peripheral targets such as the whisker pad, forepaw, and heart explants. Axon growth into these peripheral target tissues was analyzed by the use of lipophilic tracer DiI. Embryonic day 15 dorsal root and trigeminal axons grew into isochronic normal and foreign cutaneous targets. Both axon populations avoided the same age heart tissue, but grew profusely into younger (embryonic day 13) or older (postnatal) heart explants. In contrast, embryonic day 15 superior cervical or nodose ganglion axons grew heavily into the same age heart and forepaw explants and to a lesser extent into the whisker pad explants. Embryonic day 15 retinal axons grew into all three peripheral targets used in this study. Primary sensory and sympathetic axons, but not retinal axons, formed target specific patterns in the whisker pad and forepaw explants. DiI-labeling and immunostaining of primary sensory neurons in coculture revealed that these neurons retain their bipolar characteristics, and express class-specific markers such as parvalbumin, calcitonin gene-related peptide and TrkA receptors. In the whisker pad explants, axons positive for all three markers were seen to form patterns around the follicles. Our results indicate that developing peripheral targets can attract and support axon growth from a variety of sources. Whereas neurotrophins play a major role in attracting and supporting survival of subpopulations of sensory neurons, other substrate-bound or locally released molecules must regulate sensory neurite growth into specific peripheral and central targets. PMID- 9741477 TI - Cortical, thalamic, and amygdaloid connections of the anterior and posterior insular cortices. AB - Cortical, thalamic, and amygdaloid projections of the rat anterior and posterior insular cortices were examined using the anterograde transport of biocytin. Granular and dysgranular posterior insular areas between bregma and 2 mm anterior to bregma projected to the gustatory thalamic nucleus. Granular cortex projected to the subjacent dysgranular cortex which in turn projected to the agranular (all layers) and granular cortices (layers I and VI). Both granular and dysgranular posterior areas projected heavily to the dysgranular anterior insular cortex. Agranular posterior insular cortex projected to medial mediodorsal nucleus, agranular anterior insular and infralimbic cortices as well as granular and dysgranular posterior insula. No projections to the amygdala were observed from posterior granular cortex, although dysgranular cortex projected to the lateral central nucleus, dorsolateral lateral nucleus, and posterior basolateral nucleus. Agranular projections were similar, although they included medial and lateral central nucleus and the ventral lateral nucleus. Dysgranular anterior insular cortex projected to lateral agranular frontal cortex and granular and dysgranular posterior insular regions. Agranular anterior insular cortex projected to the dysgranular anterior and prelimbic cortices. Anterior insuloamygdaloid projections targeted the rostral lateral and anterior basolateral nuclei with sparse projections to the rostral central nucleus. The data suggest that the anterior insula is an interface between the posterior insular cortex and motor cortex and is connected with motor-related amygdala regions. Amygdaloid projections from the posterior insular cortex appear to be organized in a feedforward parallel fashion targeting all levels of the intraamygdaloid connections linking the lateral, basolateral, and central nuclei. PMID- 9741478 TI - Cascade projections from somatosensory cortex to the rat basolateral amygdala via the parietal insular cortex. AB - The pathways by which somatosensory information could be relayed from the cortex to the amygdaloid complex were investigated by using the anterograde axonal transport of biocytin following cortical microinjections. Injections of biocytin into head and limb areas of secondary somatosensory cortex (S2) produced heavy labeling of fibers and terminals in granular and dysgranular parietal insular cortex from bregma to 3.8 mm behind bregma but only extremely sparse labeling in the lateral and basolateral amygdaloid nuclei. Biocytin injections into granular parietal insular cortex produced a heavy labeling of the subjacent dysgranular parietal insular cortex, but only sparse labeling in the basolateral amygdala. Biocytin injections into dysgranular parietal insular cortex resulted in heavy labeling of the subjacent agranular parietal insular cortex and strong labeling of fibers and terminals in the dorsal part of lateral nucleus, with moderate labeling of fibers in the anterior and posterior basolateral nuclei, and the central nucleus. Injections into S2 labeled the ventroposterior medial, ventroposterior lateral and posterior thalamic nuclei; injections in rostral granular and dysgranular parietal insular cortex labeled the ventral posterior and parvicellular part of ventroposterior lateral thalamic nuclei; and injections in middle to caudal dysgranular parietal insular cortex labeled only the posterior nucleus. These results suggest that whereas somatosensory cortex projects only very sparsely to the amygdala, somatosensory-related inputs to the amygdala arise in the dysgranular parietal insular cortex. The association of dysgranular parietal insular cortex with the posterior thalamus suggests it may relay nociceptive information to the amygdala. PMID- 9741480 TI - Input-output connections of the "hindlimb" region of the inferior olive in cats. AB - The aim of the present study was to establish whether gracile afferents to the inferior olive are topographically organized and whether such inputs coincide with the location of cells that output to the hindlimb C1 zone in the cerebellar posterior lobe. Small injections (n=15) of the anterograde tracers Fluoro-Ruby or biotinylated dextran amine were made into gracile, resulting in anterograde labelling often distributed in partially separate, rostrocaudally directed columns within the lateral half of the contralateral rostral dorsal accessory olive (rDAO). In 12 cases, anterograde labelling was also located within the caudolateral medial accessory olive. Evidence for a topographical organization was obtained, suggesting that medial gracile targets only the most lateral part of rDAO, whereas lateral gracile also targets more medial parts. Gracile injections centred dorsally also resulted in more extensive terminal fields in rDAO than similar sized injections centred ventrally which may relate to the representation of the distal hindlimb in dorsal gracile (e.g., Millar and Basbaum, 1975). Injections of retrograde tracer (green beads) into the C1 zone in the ipsilateral caudal paramedian lobule (7 cases), resulted in retrograde cell labelling in the middle one-third of contralateral DAO in two columns that fused caudally. The proportion of labelled cells that overlapped with anterograde labelled terminals was related to the dorsoventral locus of the gracile injection site: gracile injections centred dorsally produced the largest degree of overlap and therefore, potentially, the greatest functional convergence. In summary, the results suggest that a topographical organization is present within the gracilo olivo-cerebellar projection which is discussed in relation to the functional organization of the olivocerebellar system. PMID- 9741479 TI - Immunocytochemical localization of the insulin-responsive glucose transporter 4 (Glut4) in the rat central nervous system. AB - We have previously reported that the insulin-responsive glucose transporter GLUT4 is strongly expressed by discrete areas of the rat brain (Leloup et al. [1996] Molec. Brain Res. 38:45-53). In the present study, a sensitive immunocytochemical technique has been used to analyze extensively the anatomical and ultrastructural localizations of GLUT4 in the rat central nervous system in order to gain insight into the physiological role of this transporter. We confirm that GLUT4 is expressed by numerous neurons of the brain and spinal cord, whereas glial cells are more scarcely labeled. In both light and electron microscopy, we observe that the immunoreactivity for GLUT4 is localized mainly in the somatodendritic portion of neurons, where some cisterns of rough endoplasmic reticulum, ribosomal rosettes, certain Golgi saccules, and some intracytoplasmic vesicles are labeled. In contrast, axons and nerve terminals are only occasionally immunostained in certain brain regions such as the neocortex and the ventricular surfaces for example. The GLUT4-immunoreactive structures appear concentrated and most prominently immunostained in motor areas, such as the sensorimotor cortex, most basal ganglia and related nuclei, the cerebellum and deep cerebellar nuclei, a number of reticular fields, motor nuclei of cranial nerves, and motor neurons of the ventral horn of the spinal cord. The labeled regions, which also include some sensory nuclei, are often those in which Vissing et al. ([1996] J. Cerebral Blood Flow Metab. 16:729-736) have shown that exercise stimulates local cerebral glucose utilization, so that GLUT4 might be involved in this effect. On the other hand, the fact that the anatomical localizations of GLUT4 reported here generally agree with the distribution of insulin- or insulin-receptor- related receptors is important since it indicates that the translocation of GLUT4 might also be regulated by insulin in the central nervous system. PMID- 9741481 TI - Co-grafted embryonic striatum increases the survival of grafted embryonic dopamine neurons. AB - To enhance the current therapeutic benefit of dopamine (DA) neuron grafts in Parkinson's disease, strategies must be developed that increase both DA neuron survival and fiber outgrowth into the denervated striatum. Previous work in our laboratory has demonstrated that dopaminergic neurons grow to greater size when co-grafted with striatal cell suspensions and display extensive tyrosine hydroxylase-positive (TH+) projections, but no conclusion could be reached concerning enhancement of survival of grafted DA neurons. The aim of the present study was to characterize further the potential trophic effects of striatal co grafts on grafted mesencephalic DA neuron survival. Unilaterally lesioned male Fischer 344 rats were grafted with either a suspension of mesencephalic cells or with both mesencephalic and striatal cell suspensions. Co-grafts were either mixed together or placed separately into the striatum. Lesioned rats receiving no graft served as controls. Rotational behavior was assessed following amphetamine challenge at 2 weeks prior to grafting and at 4 and 8 weeks following grafting. Only rats receiving co-grafts of nigral and striatal suspensions separated by a distance of 1 mm showed significant behavioral recovery from baseline rotational asymmetry. Both mixed and separate striatal co-grafts were associated with a doubling of DA neuron survival compared with solo mesencephalic grafts. In the mixed co-graft experiment, DA neurite branching appeared enhanced and TH-rich patches were observed, whereas with co-grafts that were separated, TH+ innervation of the intervening host striatum was increased significantly. These results provide the first evidence suggesting that nigral-striatal co-grafts, particularly those placed separately and in proximity to each other, increase both DA neuron survival and neurite extension from the mesencephalic component of the grafts. PMID- 9741482 TI - Early synaptogenesis in vitro: role of axon target distance. AB - In contrast to some previous reports suggesting a delay in synapse formation in vitro, we found that under ideal conditions, most hippocampal and hypothalamic rat neurons were synaptically coupled after 3 or 4 days in vitro. Synaptophysin immunocytochemistry revealed strongly stained presynaptic boutons by 3 days in vitro. Studies with time-lapse laser confocal imaging of FM1-43 revealed that axonal boutons were recycling their synaptic vesicles, an indication of synapse formation, as early as 3 days after plating. To test the hypothesis that neurite outgrowth was enhanced in high-density cultures, thereby increasing the probability of synapse formation, neurons were transfected with the jellyfish green fluorescent protein (GFP) gene. After 2 days in high-density cultures, green fluorescent neurites were about three times longer than in sister neurons plated in low-density cultures. Even in single dishes, GFP-transfected cells in contact with other neurons had neurites that were at least three times longer and grew faster than more isolated cells. Neurons grew longer neurites (+51%) when growing on surface membranes of heat-killed neurons than on polylysine, underlining the importance of plasma membrane contact. Calcium imaging with fura 2 and whole cell recording showed that both GABA and glutamate presynaptic release occurred after 3 or 4 days in vitro in high-density cultures but was absent in low-density cultures at this time. Together, these morphological, cytochemical, and physiological data suggest that the distance an axon must grow to find a postsynaptic partner plays a substantial role in the timing of synapse formation. Although other factors in vitro may also play a role, the distance to a postsynaptic target, which defines the interval during which an axon grows to its target, can probably account for much of the difference in timing of synapse formation previously reported in vitro. A short intercell distance may increase the concentration of limited amounts of trophic factors available to a nearby cell, and once contact is made, a neuronal membrane provides a superior substrate for neuritic elongation. PMID- 9741483 TI - Restricted expression of the neuronal intermediate filament protein plasticin during zebrafish development. AB - In the adult goldfish visual pathway, expression of the neuronal intermediate filament (nIF) protein plasticin is restricted to differentiating retinal ganglion cells (RGCs) at the margin of the retina. Following optic nerve injury, plasticin expression is elevated transiently in all RGCs coincident with the early stages of axon regeneration. These results suggest that plasticin may be expressed throughout the nervous system during the early stages of axonogenesis. To test this hypothesis, we analyzed plasticin expression during zebrafish (Danio rerio) neuronal development. By using immunocytochemistry and in situ hybridization, we found that plasticin is expressed in restricted subsets of early zebrafish neurons. Expression coincides with axon outgrowth in projection neurons that pioneer distinct axon tracts in the embryo. Plasticin is expressed first in trigeminal, Rohon-Beard, and posterior lateral line ganglia neurons, which are among the earliest neurons to initiate axonogenesis in zebrafish. Plasticin is expressed also in reticulospinal neurons and in caudal primary motoneurons. Together, these neurons establish the first behavioral responses in the embryo. Plasticin expression also coincides with initial RGC axonogenesis and progressively decreases after RGC axons reach the tectum. At later developmental stages, plasticin is expressed in a subset of the cranial nerves. The majority of plasticin-positive neurons are within or project axons to the peripheral nervous system. Our results suggest that plasticin subserves the changing requirements for plasticity and stability during axonal outgrowth in neurons that project long axons. PMID- 9741484 TI - Factor analysis and somatotyping, are these two physique classification methods comparable? AB - The aim of this study was to investigate the correspondence of physique structures estimated by the Heath-Carter anthropometric somatotyping method and a factor analysis based on the same set of 10 variables used by Heath-Carter. The investigation was carried out on a group of 200 healthy young adults of 20 years of age who were students of physical education. The mean somatotype was 2.7-4.6 3.0 for the males and 3.3-3.4-3.1 for the females. The 73% of the total variance in males and 75% in females were represented by three factors. They were identified as muscular, fatness and skeletal factors in the males, and in the females as muscular-trunk fatness, skeletal and limb fatness factors. A PCA gives different results depending on the measurements used for the calculation. The same set of variables as for the somatotyping method were used intentionally to extract the PCA factors and to evaluate the possible correspondence between these factors and the Heath-Carter components. On the basis of the correlation between the factors and the somatotype components, one can conclude that there is: (1) a high correspondence between endomorphy and fatness factors in both sexes; (2) that mesomorphy correlated positively with the muscular factor in males and negatively with the skeletal factor in both sexes; and (3) that ectomorphy was highly positively correlated with the skeletal factor and negatively with the other two factors in both sexes. Factors and somatotype components do not correspond exactly which leads to the following conclusions: (1) The three somatotype components cannot be identified as orthogonal factors in a factorial analysis based on the same measurements as for the somatotype, e.g. the ectomorphy component is not an independent factor in males or in females; (2) The muscle measurements and bone width used to estimate mesomorphy in somatotyping scored in two independent factors; and (3) The factor structure of the 10 measurements was sex dependent. PMID- 9741485 TI - The adolescent spurt and sexual maturation in girls active and not active in sport. AB - Girls actively training in sport (n=23) and girls not active in sport (n=26) were compared in terms of ages at peak height velocity (PHV) and menarche, the interval between ages at PHV and menarche, and ages at attaining stages of pubic hair and breast and the estimated duration of the stages. Subjects were longitudinally followed from about 11-18 years of age. Stature and weight were measured and stages of pubic hair and breast development were rated at approximately quarterly intervals between the initial observation and 14 years of age, at semiannual intervals until 16 years, and at irregular intervals subsequently. Age at menarche was obtained prospectively. The active girls trained 12 hours per week in rowing, track and swimming for an average of 3.9+/ 1.2 years during puberty and the growth spurt. Longitudinal stature records for individual girls were fitted with kernel regression to estimate age at PHV (years). The interval between age at PHV and age at menarche was calculated. Ages at appearance of pubic hair and breast stages 3, 4 and 5 were calculated by back interpolation, while intervals between stages 3 and 4 were calculated after log 10 transformation. Peak height velocity and menarche occur, on average, slightly later in girls active in sport, but the differences are not significant. The interval between PHV and menarche, PHV (cm/year), ages at attaining pubic hair and breast stages 3, 4 and 5, and estimated intervals between adjacent stages also do not significantly differ between girls actively training in sport and those not active in sport. Thus, regular training in sport during puberty and the adolescent spurt does not apparently influence the timing and progression of somatic and sexual maturation in girls. PMID- 9741486 TI - Changes in growth patterns in Zagreb school children related to socio-economic background over the period 1973-1991. AB - The purpose of this paper is to present the changes in growth patterns in different socio-economic classes of Zagreb school children over the period 1973 1991. Classes are defined by parental occupation. Surveys were performed in 1973, 1982 and 1991 covering 8938, 10700, and 7400 examinees aged 7 to 19 respectively. In all three observed generations boys and girls belonging to social group I (nonmanual workers' families) were taller than their peers in group III (manual workers' families). Differences were most pronounced in 1973. Mean height of children from 'mixed' families (class II) were mostly between two other groups. Positive secular changes in both genders were most pronounced in children belonging to manual workers' families--girls observed in 1991 being 2-4cm and boys 2-6cm taller than their peers in 1973. In children from nonmanual workers' families the secular increase was small in younger age groups--in boys up to 11 hardly noticeable, while in both genders from the age of 13 on, the mean height increase reached or even surpassed that observed in manual workers' children. Positive changes in all observed groups were more pronounced in the period 1973 1982 than in 1982-1991. Mean weight changes, in general over period 1973-1991 corresponded to changes in height. However, the average weight gain in girls in class I was somewhat lower compared to the gain in height, particularly in older age groups. In the same period, 19731991 the mean menarcheal age in girls showed the reversed trend i.e. a shift towards the older age. In class I the reversal was noticed in the first period, parallel to intensive height increase, while in class III positive changes in height were accompanied by significant lowering of menarcheal age. In this group the reversal was observed in the second period 1982 1991. PMID- 9741487 TI - Periodic growth in rats. AB - Microknemometry, a novel non-invasive technique, allows the accurate measurements of the lower leg length in the conscious rat, not only daily but even in periods smaller than 24 hours. Its use revealed the presence of nonlinear growth increments (mini-growth spurts) with periods between 4 and 5 days, that presented a gradual decline in amplitude when the animals were getting older, and a maximal growth rate between 0600h and 0900h. A sexual dimorphic growth pattern could be established with females growing less and presenting spurts of lower amplitude and smaller duration than males. High doses of recombinant human Growth Hormone (rhGH) stimulated growth velocity in female rates, but did not show any effect on males. Neonatal Monosodium Glutamate (MSG) treatment reduced growth both in males and females. Growth hormone (GH) replacement therapy in MSG treated animals was capable of increasing growth velocity, from day 30 onwards. The recovery was partial in males and complete in females. In intact male rats growth blockade induced by fasting was not followed by a catch up effect after refeeding, although growth velocity tended to increase and a clear catch up effect on weight was detected. Male rats seemed to grow at a maximal speed over at least the first 60 days of life, that cannot be accelerated with GH treatment, whereas female rats did respond to exogenous GH. PMID- 9741488 TI - GM and KM immunoglobulin allotypes in a Spanish Pyrenean population: Val d'Aran. AB - Four hundred and thirteen unrelated individuals (202 autochthonous and 211 non autochthonous) of Val d'Aran (Catalan Pyrenees) have been analysed for the GM and KM immunoglobulin genetic system using the inhibition haemagglutination method. This population was defined by eight GM haplotypes (GM*3 23 5*, GM*3 5*, GM*1,17 21,28, GM*1,2,17 21,28, GM*1,17 5*, GM*1,17 5,6,11,24, GM*1,17 10,11,13,15 and GM*1,17 10,11,13,15,16) inferred from the 17 observed phenotypes. The Val d'Aran population frequencies conform to Hardy-Weinberg expectations. The frequencies of phenotypes and haplotypes show a definite homogeneity between the autochthonous and non-autochthonous people of Val d'Aran and 11 other Pyrenean populations (Mauleon, Macaye, St. Jean Pied de Port, Vallee de L'Ouzom, Gavarnie, Bareges, Luz St. Sauveur, Esparros, Camurac, Capcir and Pays de Sault) that have already been studied for the same allotypes. A factorial correspondence analysis was performed for the 12 autochthonous Pyrenean populations, showing a high frequency of the GM*3 23 5* haplotype in the three Pyrenean regions (Western, Central and Eastern), while the GM*1,17 21,28 haplotype is mainly found in the Central region, GM*3 5* in the Eastern and Western zones, and the GM*1,2,17 21,28 is mainly present in the Central and Eastern populations. The results show a relative regional homogeneity, so there is no evidence of a frequency gradient in the Pyrenean populations for the GM and KM genetic systems. It may, however, be noticed that the Central Pyrenean populations form a group, with one population (Vallee de l'Ouzom) isolated from the rest, probably because of its particular model of inheritance by which the heritage is passed to the first born without sex consideration. It has been possible to point out some differences in the genetic structure of the autochthonous and non-autochthonous Val d'Aran population and to place the autochthonous Aranese group among its Pyrenean neighbours. PMID- 9741489 TI - Comparison of growth patterns of subcutaneous fat tissue in Mexican and Polish with US and Peruvian populations. AB - The development of subcutaneous fat tissue in the US was found to follow the following stages: preschool loss, prepubertal gain, adolescent loss, stabilization, adult gain, and after top fatness, age loss. 1208 subjects 2-80 years old were studied in Maya a mixed population from Merida and Progreso (Yucatan, Mexico) measuring two fatfolds. A population of 7924 from Polish towns and villages was chosen measuring 10 fatfolds. The same pattern of fatfolds growth was found in different ethnic groups, except level of thickness and age at turning points. Adolescent loss is characteristic only for boys, and stabilization for girls. The pattern was clearly expressed in females but was only slightly marked in Yucatan males and in Poles. Data was compared for triceps fatfold, and also for the summation of several fatfolds, sometimes different in the studied populations (from 2 to 10 sites was measured). Preschool loss was mainly found at 2-5 years of age, prepubertal gain at 7-10 years, adolescent loss in boys at 11-15 years, stabilization in girls at 15-20 years, adult gain at 17 45 years, top fatness in various populations occurred between 45 and 55 years of age and next age loss was observed. Most scanty fat tissue occurred in the Peru Nunoa population, next in the Poles and white and black Americans, fat tissues became more abundant in the Mexican American and Puerto Rican, and was most abundant in the Yucatecan population. US black females have thicker fatfold than white, but white males thicker than black. Differences in amount of fat tissue are probably related to nutritional levels (quantity) and habits (quality). PMID- 9741490 TI - Is the reduction of birth intervals an efficient reproductive strategy in traditional Morocco? AB - Birth interval lengths are analysed from reproductive life histories of 517 Berber peasant women of the region of Marrakesh (Southern Morocco), whose fertility developed in a full traditional context. The high mortality rates associated with short birth intervals indicate that a rapid succession of births is detrimental to the progeny. The reproductive efficiency of the traditional propensity to a large family size is therefore examined by means of two different evaluations of reproductive success: the 'absolute' reproductive success (the absolute number of offspring surviving to maturity) and the 'relative' reproductive success (the proportion of live born surviving to maturity). The first shows that close pregnancies increase the fertility rate to such an extent that the associate higher number of deaths is more than compensated for, so that the women practising short birth intervals produce more surviving offspring than the others by the end of their reproductive life. The second shows that the probability of survival is directly associated with birth interval length, the efficiency of the reproductive process being therefore greater as birth intervals grow. It is suggested that these two behaviours are not contradictory, and that they represent two successive steps of the same reproductive adjustment to evolving environmental conditions. PMID- 9741491 TI - VNTR DNA variation in the population of the island of Hvar, Croatia. AB - The aim of this study was to investigate hypervariable DNA polymorphisms in the Croatian population. Two VNTR loci, D7S22 and D12S11, were studied in a sample of 68 inhabitants of the western and eastern regions of the island of Hvar. Binned allele frequencies and heterozygosity were calculated for the two regions and measures of genetic kinship and genetic distance were computed between the two regions and between each of these regions and nine world populations. The relatively large genetic distance between the two regions of the island of Hvar (0.0353) may reflect historical immigrations from the mainland and interesting peculiarities of the populations that arrived from the Balkan peninsula. PMID- 9741492 TI - Calculating components of the coefficient of relationship. AB - The coefficient of relationship by isonymy is Ri=sigma(n(n-1)/2(N(N-1)) in which n is the number of persons of each surname and N=sigma n. Dividing Ri into two components, one for the contribution of co-residence (family size) and the other for diversity of surnames among residences is achieved by letting a1 represent sigma n(n-1) for all residents, a2 represent sigma n(n-1) after eliminating all but one individual of any name at any residence, and b1 represent 2N(N-1) for all residents. Then the component for inter-residence diversity is a2/b1 and the component for relationship by co-residence (including the interaction) is (a1 a2)/b1. By the same logic it is possible to calculate separately the interaction component, but the additional information seems of limited importance. PMID- 9741493 TI - Left ventricular pseudoaneurysm. AB - Left ventricular (LV) pseudoaneurysms form when cardiac rupture is contained by adherent pericardium or scar tissue. Although LV pseudoaneurysms are not common, the diagnosis is difficult and they are prone to rupture. We evaluated the clinical presentation, diagnostic accuracy of imaging modalities, results of therapy and prognosis of 290 patients with LV pseudoaneurysms. Most cases of LV pseudoaneurysm were related to myocardial infarction (particularly inferior wall myocardial infarction) and cardiac surgery. Congestive heart failure, chest pain and dyspnea were the most frequently reported symptoms, but >10% of patients were asymptomatic. Physical examination revealed a murmur in 70% of patients. Almost all patients had electrocardiographic abnormalities, but these were usually nonspecific ST segment changes; only 20% of patients had ST segment elevation. Although radiographic findings were also usually nonspecific, the appearance of a mass was present in more than one half of patients and may be an important clue to the correct diagnosis. Left ventricular angiography was the most definitive test and can be useful in planning surgery since concomitant coronary angiography can be performed. Regardless of treatment, patients with LV pseudoaneurysms had a high mortality rate, especially those who did not undergo surgery. Because the symptoms, signs, electrocardiographic abnormalities and radiographic findings seen in patients with LV pseudoaneurysms can be indistinguishable from those in patients with coronary disease alone, a high clinical index of suspicion is needed to avoid missing the diagnosis. PMID- 9741494 TI - Biocompatibility aspects of new stent technology. AB - Stent implantation represents a major step forward since the introduction of coronary angioplasty. As indications continue to expand, better understanding of the early and late biocompatibility issues appears critical. Persisting challenges to the use of intracoronary stents include the prevention of early thrombus formation and late neointima development. Different metals and designs have been evaluated in animal models and subsequently in patients. Polymer coatings have been proposed to improve the biocompatibility of metallic stents or to serve as matrix for drug delivery and they are currently undergoing clinical studies. The promises of a biodegradable stent have not yet been fulfilled although encouraging results have recently been reported. Continuous low dose rate brachytherapy combining the scaffolding effect of the stent with localized radiation therapy has witnessed the development and early clinical testing of radioactive stents. The combined efforts of basic scientists and clinicians will undoubtedly contribute to the improvement of stent biocompatibility in the future. PMID- 9741495 TI - Stenting in acute coronary syndromes: a comparison of radial versus femoral access sites. AB - OBJECTIVES: The purpose of the present study was to compare the radial approach with the femoral approach for coronary stenting in patients with acute coronary syndromes. BACKGROUND: Aggressive anticoagulation in patients with acute coronary syndromes increases the risk of femoral vascular complications. The transradial approach has the potential to significantly reduce the incidence of access site bleeding complications in this group of patients. METHODS: One hundred forty-two patients with acute coronary syndromes undergoing coronary stenting were prospectively randomized to have their procedure performed from either the radial or femoral access site and the results compared. RESULTS: Nine of 74 patients randomized to the radial group crossed over to the femoral group (6 negative Allen tests, 3 access failures). Patient demographics were the same in both groups. Primary success was identical: 96% radial, 96% femoral, ns. There were no procedural myocardial infarctions or deaths, and no patient was referred for emergency bypass surgery. There were no access site bleeding complications in the radial group as opposed to 3 (4%) in the femoral group, p < 0.01. Postprocedure length of stay, days (1.4+/-0.2 radial vs. 2.3+/-0.4 femoral, p < 0.01) as well as total hospital length of stay (3.0+/-0.3 radial vs. 4.5+/-0.5 femoral, p < 0.01) were significantly reduced in the radial group. Total hospital charge was also significantly lower in the radial group ($20,476+/-811 radial versus $23,389+/-1,180 femoral, p < 0.01). CONCLUSION: Coronary stenting from the radial approach is efficacious in patients with acute coronary syndromes. Access site bleeding complications are less, and early ambulation results in a shorter hospital length of stay. There was a 15% reduction in total hospital charge in the radial group. PMID- 9741496 TI - Coronary artery stenting in the elderly: short-term outcome and long-term angiographic and clinical follow-up. AB - OBJECTIVES: This study sought to compare the short- and long-term outcomes of elderly patients undergoing coronary artery stenting with those of younger patients and to determine the long-term clinical outcome and survival of elderly patients post stent implantation. BACKGROUND: Elderly patients undergoing coronary revascularization are considered a high-risk group. Few data exist that relate the results of stenting in treating coronary artery disease in the elderly population. METHODS: All elderly patients >75 years of age who underwent coronary artery stenting between March 1993 and July 1997 (n=137) at our center were compared to the patients <75 who underwent coronary artery stenting during the same time period (n=2,551). Long-term clinical follow-up and survival were determined for the elderly group. RESULTS: Elderly patients presented with lower ejection fractions (54% vs. 58%, p=0.0001), more unstable angina (47% vs. 28%, p=0.0001), and more multivessel disease (78% vs. 62%, p= 0.0001) than younger patients. These older patients had higher rates of procedure related complications including procedural myocardial infarction (MI) (2.9% vs. 1.7%, p=0.2), emergency CABG (3.7% vs. 1.4%, p=0.04), and death (2.2% vs. 0.12%, p=0.0001). Angiographic follow-up, obtained in both groups, demonstrated significantly higher restenosis rates in the elderly versus younger patients (47% vs. 28%, p=0.0007). Longer term clinical follow-up, which was obtained only in the elderly group, showed that at a mean follow-up period of 12 months post coronary stenting, elderly survival free from death, MI, revascularization and angina was 54% and that their overall survival was 91%. Subanalysis of the elderly patients who died showed much higher incidence of combined unstable angina (80%), prior MI (60%), lower ejection fraction (46%), multivessel disease (100%) and complex lesions (100%) than the overall group. CONCLUSIONS: Elderly patients who undergo coronary artery stenting have significantly higher rates of procedural complications and worse six month outcomes than younger patients, especially those who present with combined unstable angina, history of MI, EF < 50%, multivessel disease and complex lesions. Overall survival in the elderly population at 12 months postcoronary artery stenting was 91% and event-free survival was 54%. PMID- 9741497 TI - The influence of diabetes mellitus on acute and late clinical outcomes following coronary stent implantation. AB - OBJECTIVES: We compared the clinical outcomes following coronary stent implantation in insulin-treated diabetes mellitus (IDDM), non-IDDM patients, and nondiabetic patients. BACKGROUND: Diabetic patients have increased restenosis and late morbidity following balloon angioplasty. The impact of diabetes mellitus (DM), especially IDDM, on in-stent restenosis is not known. METHODS: We studied 954 consecutive patients with native coronary artery lesions treated with elective Palmaz-Schatz stents implantation using conventional coronary angiographic and intravascular ultrasound methodology. Procedural success, major in-hospital complications, and 1-year clinical outcome were compared according to the diabetic status. RESULTS. In-hospital mortality was 2% in IDDM, significantly higher (p <0.02) compared with non-IDDM (0%) and nondiabetics (0.3%). Stent thrombosis did not differ among groups (0.9% in IDDM vs. 0% in non-IDDM and 0% in nondiabetics, p >0.1). During follow-up, target lesion revascularization (TLR) was 28% in IDDM, significantly higher (p <0.05) compared with non-IDDM (17.6%) and nondiabetics (16.3%). Late cardiac event-free survival (including death, myocardial infarction [MI], and any coronary revascularization procedure) was significantly lower (p=0.0004) in IDDM (60%) compared with non-IDDM (70%) and nondiabetic patients (76%). By multivariate analysis, IDDM was an independent predictor for any late cardiac event (OR=2.05, p=0.0002) in general and TLR (odds ratio=2.51, p=0.0001) in particular. CONCLUSIONS. In a large consecutive series of patients treated by elective stent implantation, IDDM patients were at higher risk for in-hospital mortality and subsequent TLR and, as a result, had a significantly lower cardiac event-free survival rate. On the other hand, acute and long-term procedural outcome was found to be similar for non-IDDM compared with nondiabetic patients. PMID- 9741498 TI - Long-term analysis of conventional coronary balloon angioplasty and an initial "stent-like" result. The NHLBI PTCA Registry. AB - OBJECTIVES: We examined the influence of an initial "stent-like" result on long term outcome in patients in the 1985-86 NHLBI PTCA Registry. BACKGROUND: Stent use in selected patients is associated with improved angiographic and short-term clinical outcome; however, due to potential for in-stent restenosis and high costs of stents, there is interest in a strategy of more optimal dilatation to achieve a "stent-like" result without a stent. The long-term outcome of patients with a "stent-like" percutaneous transluminal coronary angioplasty (PTCA) remains unknown. METHODS: Ten-year outcome was compared between 225 successfully treated patients with and 1,764 successfully treated patients without an initial "stent like" result ( > or = 1 lesion dilated to < or = 10% stenosis). The sample had 75% and 80% power, respectively, to detect an absolute difference of 8% in the 10 year rate of death and myocardial infarction (MI) between the two groups. RESULTS: Ten-year rates of death and MI were similar between the stent-like and non-stent-like groups (22.3% vs. 22.2%, 17.6% vs. 17.9%), however, there was less target lesion revascularization in the stent-like group (30.2% vs. 36.8%). In subgroup analysis of patients with multivessel disease, those with a stent-like result had less follow-up bypass surgery (25.2% vs. 32.7%), yet more repeat PTCA (53.8% vs. 42.7%). These findings were unaffected by adjustment for differences in baseline characteristics between the two patient groups. CONCLUSIONS: Achievement of an initial stent-like result via balloon angioplasty alone may not appreciably reduce the long-term risk of death or MI, nor confer equivalent clinical benefit as achieving a stent-like result with a stent. PMID- 9741499 TI - A prospective randomized trial of triage angiography in acute coronary syndromes ineligible for thrombolytic therapy. Results of the medicine versus angiography in thrombolytic exclusion (MATE) trial. AB - OBJECTIVES: The purpose of this study was to determine if early triage angiography with revascularization, if indicated, favorably affects clinical outcomes in patients with suspected acute myocardial infarction who are ineligible for thrombolysis. BACKGROUND: The majority of patients with acute myocardial infarction and other acute coronary syndromes are considered ineligible for thrombolysis and therefore are not afforded the opportunity for early reperfusion. METHODS: This multicenter, prospective, randomized trial evaluated in a controlled fashion the outcomes following triage angiography in acute coronary syndromes ineligible for thrombolytic therapy. Eligible patients (n=201) with <24 h of symptoms were randomized to early triage angiography and subsequent therapies based on the angiogram versus conventional medical therapy consisting of aspirin, intravenous heparin, nitroglycerin, beta-blockers, and analgesics. RESULTS: In the triage angiography group, 109 patients underwent early angiography and 64 (58%) received revascularization, whereas in the conservative group, 54 (60%) subsequently underwent nonprotocol angiography in response to recurrent ischemia and 33 (37%) received revascularization (p=0.004). The mean time to revascularization was 27+/-32 versus 88+/-98 h (p=0.0001) and the primary endpoint of recurrent ischemic events or death occurred in 14 (13%) versus 31 (34%) of the triage angiography and conservative groups, respectively (45% risk reduction, 95% CI 27-59%, p=0.0002). There were no differences between the groups with respect to initial hospital costs or length of stay. Long-term follow-up at a median of 21 months revealed no significant differences in the endpoints of late revascularization, recurrent myocardial infarction, or all cause mortality. CONCLUSIONS: Early triage angiography in patients with acute coronary syndromes who are not eligible for thrombolytics reduced the composite of recurrent ischemic events or death and shortened the time to definitive revascularization during the index hospitalization. Despite more frequent early revascularization after triage angiography, we found no long-term benefit in cardiac outcomes compared with conservative medical therapy with revascularization prompted by recurrent ischemia. PMID- 9741501 TI - Role of different determinants of psychological distress in acute coronary syndromes. AB - OBJECTIVES: The aim of this study was to examine the prevalence of psychological distress and of its major determinants in acute coronary patients and in a control group. BACKGROUND: The prevalence and major determinants of psychological distress in acute coronary patients are not clear. METHODS: One hundred and thirty cardiac patients (110 men, age 56+/-9; 85 with acute myocardial infarction and 45 with unstable angina) and 102 controls hospitalized for acute trauma (70 men, age 55+/-9 years) were studied and the level of psychological distress estimated by a Modified Maastricht Questionnaire, self-ratings and ratings by a close relative. Major determinants of psychological distress were assessed by the Life Events Assessment, the Social Support Questionnaire and the Ways of Coping Checklist. RESULTS: The average level of psychological distress was significantly higher (p < 0.001) in coronary patients than in controls in all tests (self evaluation=7.1+/-2.3 vs 4.3+/-2.4; relative-evaluation = 7.4+/-2.4 vs 4.2+/- 2.5; Modified Maastricht Questionnaire=91+/-32 vs 59+/-30). Cardiac patients reported significantly higher (p < 0.05) levels of social isolation (28.9+/-11.1 vs 23.4+/ 8.8), self-blame (7.2+/-1.9 vs 5.8+/- 1.6) and avoidance (21.1+/-3.5 vs 18.9+/-3) and more painful life events (3.9+/-3.8 vs 2.6+/-2.2), than controls. However, not all patients had evidence of distress; indeed, cluster analysis identified a subgroup that comprised 75% of controls and 25% of cardiac patients with no determinants eliciting distress, while the other four subgroups, with one or more determinants of distress, comprised about 75% of patients and only 25% of controls. CONCLUSIONS: These results show that a high level of psychological distress is detectable in about 75% of patients with acute myocardial infarction or unstable angina and is related to one or more major determinants. PMID- 9741500 TI - Mast cell infiltration in acute coronary syndromes: implications for plaque rupture. AB - OBJECTIVES: To define the role of mast cells in plaque destabilization. BACKGROUND: Inflammation is an essential feature of human coronary plaques. Macrophages and T lymphocytes are considered to contribute to destabilization of the plaques. The role of mast cells in this setting is less well studied. We therefore counted the mast cells in coronary atherectomy specimens from patients with chronic stable angina, unstable angina and severe unstable angina. METHODS: Patients studied had chronic stable angina (group 1, n=11), "stabilized" unstable angina (group 2; Braunwald's class I and II, n=11) and "refractory" unstable angina (group 3; Braunwald's class III, n=7). Samples of culprit lesions (n=29) were obtained by directional atherectomy, snap-frozen and analyzed immunohistochemically. The numbers of mast cells and T lymphocytes per square millimeter squared were counted and the areas containing macrophages and smooth muscle cells were measured by computed planimetry. RESULTS: We found that the numbers of mast cells and T lymphocytes increased from group 1 through groups 2 to 3. Specimens from group 3 also contained the largest numbers of tumor necrosis factor alpha (TNF-alpha)-positive mast cells and of matrix metalloproteinase (MMP)-9 (92 kD gelatinase)-positive macrophages. CONCLUSIONS: Unstable coronary syndromes are associated with increased numbers of mast cells in culprit lesions. Activated mast cells secrete neutral proteases capable of degrading the extracellular matrix and TNF-alpha, capable of stimulating macrophages to synthesize MMP-9. Our observations suggest that mast cells, in addition to macrophages, contribute to matrix degradation and, hence, to progression of coronary syndromes. PMID- 9741502 TI - Intravenous diltiazem in acute myocardial infarction. Diltiazem as adjunctive therapy to activase (DATA) trial. AB - OBJECTIVES: This study was defined as a pilot investigation of the usefulness and safety of intravenous diltiazem as adjunctive therapy to tissue plasminogen activator in acute myocardial infarction, followed by oral therapy for 4 weeks. BACKGROUND: Experimental studies have documented that calcium antagonists protect the myocardial cell against the damage caused by coronary artery occlusion and reperfusion, yet no benefits have been conclusively demonstrated in acute myocardial infarction (AMI) in humans. METHODS: In this pilot study, 59 patients with an AMI treated with tissue-type plasminogen activator (t-PA) were randomized, double blinded, to intravenous diltiazem or placebo for 48 h, followed by oral therapy for 4 weeks. The primary objective was to detect an effect on indices of regional left ventricular function and perfusion. Patients were also closely monitored for clinical events, coronary artery patency and indices of infarct size and of left ventricular function. RESULTS: Creatine kinase elevation, Q wave score, global and regional left ventricular function and coronary artery patency at 48 h were not significantly different between the diltiazem and placebo groups. A greater improvement observed in regional perfusion and function with diltiazem was likely explained by initial larger defects. Diltiazem, compared to placebo, reduced the rate of death, reinfarction or recurrent ischemia at 35 days from 41% to 13% (p=0.027) and prevented the need for an urgent intervention. The rate of death or myocardial infarction was reduced by 65% (p=0.15). These benefits could not be explained by differences in baseline characteristics such as age, site and extent of infarction, time of inclusion or concomitant therapy. Heart rate and blood pressure were reduced throughout the study with active diltiazem treatment. Side effects of diltiazem were bradycardia and hypotension that required transient or permanent discontinuation of the study drug in 27% of patients, vs. 17% of patients with placebo. CONCLUSIONS: A protective effect for clinical events related to early postinfarction ischemia and reinfarction was suggested in this study, with diltiazem administered intravenously with t-PA followed by oral therapy for 1 month, with no effect on coronary artery patency and left ventricular function and perfusion. PMID- 9741503 TI - Influence of treatment delay on infarct size and clinical outcome in patients with acute myocardial infarction treated with primary angioplasty. AB - OBJECTIVES: The purpose of this analysis was to determine the influence of an additional treatment delay inherent in transfer to an angioplasty center for primary angioplasty of patients with acute myocardial infarction who are first admitted to hospitals without angioplasty facilities. BACKGROUND: Several randomized trials have demonstrated the benefits of primary angioplasty in acute myocardial infarction. In recent years, increasing numbers of patients with myocardial infarction, initially admitted to hospitals without angioplasty facilities are transported to our hospital for primary angioplasty. However, the additional delay due to the transport may have a deleterious effect on infarct size and clinical outcome. METHODS: In a three-year period (December 1993 to November 1996), 207 consecutive patients who were transferred for primary angioplasty were analyzed in a matched comparison with non-transferred patients. Matching criteria were age, sex, infarct location, presentation delay and Killip class. RESULTS: Patients who were transferred had an additional median delay of 43 min. This resulted in a more extensive enzymatic infarct size and a lower ejection fraction measured at 6 months. The rate of angioplasty success defined as TIMI grade 3 flow, and the 6-month mortality rate (7%) were comparable in both groups. CONCLUSIONS: The additional delay had a deleterious effect on myocardial salvage, reflected by a larger infarct size and a lower left ventricular function. However, the patency rate and 6-month clinical outcome were not affected by this delay. PMID- 9741504 TI - Atenolol use and clinical outcomes after thrombolysis for acute myocardial infarction: the GUSTO-I experience. Global Utilization of Streptokinase and TPA (alteplase) for Occluded Coronary Arteries. AB - OBJECTIVES: We assessed the use and effects of acute intravenous and later oral atenolol treatment in a prospectively planned post hoc analysis of the GUSTO-I dataset. BACKGROUND: Early intravenous beta blockade is generally recommended after myocardial infarction, especially for patients with tachycardia and/or hypertension and those without heart failure. METHODS: Besides one of four thrombolytic strategies, patients without hypotension, bradycardia or signs of heart failure were to receive atenolol 5 mg intravenously as soon as possible, another 5 mg intravenously 10 min later and 50 to 100 mg orally daily during hospitalization. We compared the 30-day mortality of patients given no atenolol (n=10,073), any atenolol (n=30,771), any intravenous atenolol (n=18,200), only oral atenolol (n=12,545) and both intravenous and oral drug (n=16,406), after controlling for baseline differences and for early deaths (before oral atenolol could be given). RESULTS: Patients given any atenolol had a lower baseline risk than those not given atenolol. Adjusted 30-day mortality was significantly lower in atenolol-treated patients, but patients treated with intravenous and oral atenolol treatment vs. oral treatment alone were more likely to die (odds ratio, 1.3; 95% confidence interval, 1.0 to 1.5; p=0.02). Subgroups had similar rates of stroke, intracranial hemorrhage and reinfarction, but intravenous atenolol use was associated with more heart failure, shock, recurrent ischemia and pacemaker use than oral atenolol use. CONCLUSIONS: Although atenolol appears to improve outcomes after thrombolysis for myocardial infarction, early intravenous atenolol seems of limited value. The best approach for most patients may be to begin oral atenolol once stable. PMID- 9741505 TI - Clinical predictors of early infarct-related artery patency following thrombolytic therapy: importance of body weight, smoking history, infarct-related artery and choice of thrombolytic regimen: the GUSTO-I experience. Global Utilization of Streptokinase and t-PA for Occluded Coronary Arteries. AB - OBJECTIVES: The purpose of this study was to determine patient characteristics that are a priori predictors of early infarct related artery patency following thrombolytic therapy, and to provide a paradigm which may identify patients who would be most likely to achieve restoration of normal (TIMI 3) coronary flow in response to thrombolytic therapy. BACKGROUND: Restoration of infarct-related artery perfusion in acute myocardial infarction is necessary for preservation of ventricular function and mortality reduction. Clinical variables that are a priori predictors of early patency with currently available thrombolytic regimens have not been fully characterized. METHODS: The probability of early infarct related artery patency (TIMI 3 flow) was determined by multivariable logistic regression. We determined a reduced (parsimonious) model for predicting early (90 min) infarct-related artery patency (TIMI grade 3) based on data from 1,030 patients in the GUSTO-I Angiographic study. RESULTS: Predictors of 90 min TIMI 3 flow are use of an accelerated t-PA regimen (vs. streptokinase containing regimens) (chi2=39.1; p < or = 0.0001), infarct related artery (RCA/Lcx vs. LAD) (chi2=12.7; p=0.0004), body weight (chi2=10.3; p=0.001) and history of smoking (chi2=7.4; p=0.007). Time from symptom onset to treatment was not significant (p=0.71). CONCLUSIONS: The efficacy of currently available thrombolytic regimens is chiefly dependent on choice of thrombolytic regimen, body weight, infarct related coronary artery and smoking history. Clinical variables alone correctly predict a priori TIMI 3 flow in the infarct-related artery 64% of the time. Patients with body weights greater than 85 kg are at a significant disadvantage with regard to achieving successful thrombolysis compared to those with lesser body weights. PMID- 9741506 TI - Significance of rest technetium-99m sestamibi imaging for the prediction of improvement of left ventricular dysfunction after Q wave myocardial infarction: importance of infarct location adjusted thresholds. AB - OBJECTIVE: The value of rest technetium-99m (Tc-99m) sestamibi scintigraphy under oral nitrate medication to predict myocardial viability was examined in patients with chronic infarcts. BACKGROUND: The value of rest Tc-99m sestamibi to predict viability in infarct regions has not been fully established because significant underestimation of viability, especially in the inferior myocardial wall, has been reported. METHODS: Forty patients with Q wave myocardial infarction underwent Tc-99m sestamibi single-photon emission computed tomography under nitrate medication before revascularization of the infarct-related artery. Wall motion was quantified from paired angiograms before and 4 months after revascularization. Tracer uptake was quantified in the central infarct region identified on the angiogram. RESULTS: The average Tc-99m sestamibi uptake in the central infarct region of patients with anterior infarcts and improvement of left ventricular function was significantly higher (68+/-12%, mean+/-SD) than in patients without improvement of function (40+/-14%, p < 0.02). The average Tc-99m sestamibi uptake in the central infarct region of patients with improvement of function and inferior infarcts was significantly lower (43+/-7%) than in patients with anterior infarcts (68+/-12%, p < 0.003), but was significantly higher than in patients with inferior infarction and no improvement of function (31+/-7%, p < 0.02). Using an infarct location adjusted optimal threshold (50% for anterior infarcts, 35% for inferior infarcts), Tc-99m sestamibi had a positive predictive value of 90% and a negative predictive value of 91% for improvement of left ventricular function. CONCLUSION: Quantitative rest Tc-99m sestamibi scintigraphy after oral nitrates reliably predicts improvement of left ventricular function after revascularization if infarct location adjusted thresholds are used. PMID- 9741508 TI - Plaque size, vessel size and plaque vulnerability: bigger may not be better. PMID- 9741507 TI - Relation of arterial geometry to luminal narrowing and histologic markers for plaque vulnerability: the remodeling paradox. AB - OBJECTIVE: To relate local arterial geometry with markers that are thought to be related to plaque rupture. BACKGROUND: Plaque rupture often occurs at sites with minor luminal stenosis and has retrospectively been characterized by colocalization of inflammatory cells. Recent studies have demonstrated that luminal narrowing is related with the mode of atherosclerotic arterial remodeling. METHODS: We obtained 1,521 cross section slices at regular intervals from 50 atherosclerotic femoral arteries. Per artery, the slices with the largest and smallest lumen area, vessel area and plaque area were selected for staining on the presence of macrophages (CD68), T-lymphocytes (CD45RO), smooth muscle cells (alpha-actin) and collagen. RESULTS: Inflammation of the cap or shoulder of the plaque was observed in 33% of all cross sections. Significantly more CD68 and CD45RO positive cells, more atheroma, less collagen and less alpha-actin positive staining was observed in cross sections with the largest plaque area and largest vessel area vs. cross sections with the smallest plaque area and smallest vessel area, respectively. No difference in the number of inflammatory cells was observed between cross sections with the largest and smallest lumen area. CONCLUSION: Intraindividually, pathohistologic markers previously reported to be related to plaque vulnerability were associated with a larger plaque area and vessel area. In addition, inflammation of the cap and shoulder of the plaque was a common finding in the atherosclerotic femoral artery. PMID- 9741510 TI - Comparison of electron beam computed tomography scanning and conventional risk factor assessment for the prediction of angiographic coronary artery disease. AB - OBJECTIVE: To determine whether electron beam computed tomography (CT) adds to conventional risk factor assessment in the prediction of angiographic coronary artery disease. BACKGROUND: Electron beam CT scanning can be used to predict the severity of coronary atherosclerosis, but whether it does so independently of conventional risk factors is unclear. METHODS: Electron beam CT scans were performed and conventional risk factors were measured in 290 men and women undergoing coronary arteriography for clinical indications. The association of the electron beam CT-derived coronary artery calcium score and conventional risk factors with the presence and severity of angiographically defined coronary atherosclerosis was analyzed by logistic regression and receiver-operator characteristics analysis. RESULTS: Age, the ratio of total cholesterol to high density lipoprotein (HDL) cholesterol and the coronary calcium score were significantly and independently associated with the presence of any coronary disease and obstructive coronary disease. In association with any coronary disease, odds ratios for age, the ratio of total cholesterol to HDL cholesterol and calcium score, highest quartile vs. lowest quartile, were 6.01 (95% confidence interval 2.87 to 12.56), 3.14 (1.56 to 6.31) and 94.08 (21.06 to 420.12), respectively. For obstructive coronary disease, highest quartile vs. lowest quartile, the respective odds ratios for age, the ratio of total cholesterol to HDL and calcium score were 3.86 (1.86 to 8.00), 4.11 (1.98 to 8.52) and 34.12 (12.67 to 91.86). Male gender was also significantly associated with any coronary disease (odds ratio 2.19, p=0.04) and obstructive coronary disease (odds ratio 2.07, p=0.04). Cigarette smoking was significantly associated with any coronary disease (odds ratio=2.74, p=0.004), and diabetes was significantly associated with obstructive disease (odds ratio 3.16, p=0.01). After adjustment for the coronary calcium score and other risk factors, it was determined that triglycerides, family history and hypertension were not significantly associated with any disease state. A coronary calcium score >80 (Agatston method) was associated with an increased likelihood of any coronary disease regardless of the number of risk factors, and a coronary calcium score > or = 170 was associated with an increased likelihood of obstructive coronary disease regardless of the number of risk factors (p < 0.001). CONCLUSIONS: Electron beam CT scanning offers improved discrimination over conventional risk factors in the identification of persons with any angiographic coronary disease or angiographic obstructive coronary disease. PMID- 9741509 TI - Treating patients with documented atherosclerosis to National Cholesterol Education Program-recommended low-density-lipoprotein cholesterol goals with atorvastatin, fluvastatin, lovastatin and simvastatin. AB - OBJECTIVES: This study compared the efficacy and safety of atorvastatin, fluvastatin, lovastatin, and simvastatin in patients with documented atherosclerosis treated to U.S. National Cholesterol Education Program (NCEP) recommended low-density-lipoprotein (LDL) cholesterol concentration (< or = 100 mg/dl [2.59 mmol/liter]). BACKGROUND: For patients with advanced atherosclerosis, NCEP recommends lipid-lowering drug therapy if LDL cholesterol remains > or = 130 mg/dl (3.36 mmol/liter). METHODS: A total of 318 men or women with documented atherosclerosis and LDL cholesterol > or = 130 mg/dl (3.36 mmol/liter) and < or = 250 mg/dl (6.5 mmol/liter), and triglycerides < or = 400 mg/dl (4.5 mmol/liter) participated in this 54-week, multicenter, open-label, randomized, parallel group, active-controlled, treat-to-target study. Patients were titrated at 12 week intervals until the LDL cholesterol goal was reached. Number of patients reaching target LDL cholesterol levels and dose to reach target were evaluated. RESULTS: At the starting doses, atorvastatin 10 mg produced significantly greater decreases (p < 0.05) in plasma LDL cholesterol than the other treatments. Subsequently, the percentage of patients reaching goal at the starting dose was 32% for atorvastatin, 1% for fluvastatin, 10% for lovastatin and 22% for simvastatin. Atorvastatin-treated patients required a lower median dose than other treatments. Median doses at week 54 with the last available visit carried forward were atorvastatin 20 mg/day, fluvastatin 40 mg/day + colestipol 20 g/day, lovastatin 80 mg/day, simvastatin 40 mg/day. CONCLUSIONS: A significantly greater number (p < 0.05) of patients with confirmed atherosclerosis treated with atorvastatin reached the target LDL cholesterol concentration at the starting dose than patients treated with fluvastatin or lovastatin, and significantly fewer (p < 0.05) patients treated with atorvastatin required combination therapy with colestipol to achieve target LDL cholesterol concentrations than all other statins tested. PMID- 9741511 TI - Prognostic value of ischemic electrocardiographic findings for cardiovascular mortality in men and women. AB - OBJECTIVES: The aim of this study was to investigate the independent prognostic value of ischemic electrocardiographic (ECG) findings for cardiovascular mortality and to evaluate a possible sex-differential in this regard. BACKGROUND: In previous reports, ST segment and T wave changes on the resting ECG were described as independent risk factors for development of coronary heart disease. Although more prevalent in women, they are often given less clinical importance than in men. METHODS: Ten-year follow-up data from the Belgian Interuniversity Research on Nutrition and Health study were used. The results presented here are based on ECGs of the 4,797 men and 4,320 women, aged 25 to 74 years, who were free of angina pectoris at the start of follow-up, had no history of myocardial infarction (MI) and showed no Q wave evidence of an old MI on their ECG. RESULTS: At baseline, the age-standardized prevalence of an "ischemic ECG" (Minnesota codes I3, IV1-3, V1-3 or VII1) was 8.4% in men and 10.6% in women. Cardiovascular mortality rates in men and women with an ischemic ECG were respectively 7.7 and 2.6 per 1,000 person-years, compared with 2.3 and 1.0 in those with no such ECG findings. After correction for the potential confounding effects of established cardiovascular disease (CVD) risk factors, the multivariately adjusted risk ratios were 2.45 (95% confidence interval [CI]: 1.70 to 3.53) for men and 2.16 (95% CI: 1.30 to 3.58) for women. Testing the interaction between an ischemic ECG and sex on CVD mortality revealed that the risk ratios were not significantly changed (p=0.95). The etiologic fraction of CVD deaths attributable to an ischemic ECG was estimated as 19.3% for men and 22.4% for women. Both men and women with major ischemic findings in their baseline electrocardiogram (Minnesota codes IV1,2, V1,2 or VII1) had a fourfold increased risk of CVD death. CONCLUSION: These results support the hypothesis that women with ischemic ECG findings are at the same increased risk for CVD mortality as men. PMID- 9741512 TI - Superiority of "triple" drug therapy in heart failure: insights from the PROVED and RADIANCE trials. Prospective Randomized Study of Ventricular Function and Efficacy of Digoxin. Randomized Assessment of Digoxin and Inhibitors of Angiotensin-Converting Enzyme. AB - OBJECTIVES: We sought to study the efficacy of "triple" therapy with digoxin, diuretic and angiotensin-converting enzyme inhibitor (ACEI) compared to other combinations of these drugs in patients with symptomatic left ventricular systolic dysfunction. BACKGROUND: Controversy continues concerning the role of combining digoxin with diuretic and ACEI in the initial management of patients with heart failure. METHODS: The study utilized data from two studies of digoxin efficacy: Prospective Randomized Study of Ventricular Function and Efficacy of Digoxin (PROVED) and Randomized Assessment of Digoxin and Inhibitors of Angiotensin-Converting Enzyme (RADIANCE). Worsening heart failure defined as augmentation of heart failure therapy or an emergency room visit or hospitalization for increased heart failure was the main outcome measure. RESULTS: A total of 266 patients comprising the four treatment groups of the combined PROVED (diuretic alone or digoxin and diuretic) and RADIANCE (ACEI and diuretic, or digoxin, diuretic and ACEI) trials were analyzed. Worsening heart failure occurred in only 4 of the 85 patients who continued digoxin, diuretic and ACEI therapy (4.7%) compared to 18 of the 42 patients (19%) on digoxin and diuretic therapy (p=0.009), to 23 of the 93 patients (25%) on ACEI and diuretic therapy (p=0.001) and to 18 of the 46 patients (39%) on diuretic alone (p < 0.001). Life table and multivariate analysis also demonstrated that worsening heart failure was least likely in patients treated with triple therapy (p < 0.01 vs. all other groups). CONCLUSION: Pending definitive, prospective clinical trials, our results argue for triple therapy as the initial management of patients with symptomatic heart failure due to systolic dysfunction. PMID- 9741513 TI - Triple drug therapy--what is next? PMID- 9741514 TI - Atrial fibrillation is associated with an increased risk for mortality and heart failure progression in patients with asymptomatic and symptomatic left ventricular systolic dysfunction: a retrospective analysis of the SOLVD trials. Studies of Left Ventricular Dysfunction. AB - OBJECTIVE: This study undertook to determine if the presence of atrial fibrillation in patients with asymptomatic and symptomatic left ventricular dysfunction was associated with increased mortality and, if so, whether the increase could be attributed to progressive heart failure or arrhythmic death. BACKGROUND: Atrial fibrillation is a common condition in heart failure with the potential to impact hemodynamics and progression of left ventricular systolic dysfunction as well as the electrophysiologic substrate for arrhythmias. The available data do not conclusively define the effect of atrial fibrillation on prognosis in heart failure. METHODS: A retrospective analysis of the Studies of Left Ventricular Dysfunction Prevention and Treatment Trials was conducted that compared patients with atrial fibrillation to those in sinus rhythm at baseline for the risk of all-cause mortality, progressive pump-failure death and arrhythmic death. RESULTS: The patients with atrial fibrillation at baseline, compared to those in sinus rhythm, had greater all-cause mortality (34% vs. 23%, p < 0.001), death attributed to pump-failure (16.7% vs. 9.4%, p < 0.001) and were more likely to reach the composite end point of death or hospitalization for heart failure (45% vs. 33%, p < 0.001), but there was no significant difference between the groups in arrhythmic deaths. After multivariate analysis, atrial fibrillation remained significantly associated with all-cause mortality (relative risk [RR] 1.34, 95% confidence interval [CI] 1.12 to 1.62, p=0.002), progressive pump-failure death (RR 1.42, 95% CI 1.09 to 1.85, p=0.01), the composite end point of death or hospitalization for heart failure (RR 1.26, 95% CI 1.03 to 1.42, p=0.02), but not arrhythmic death (RR 1.13; 95% CI 0.75 to 1.71; p=0.55). CONCLUSIONS: The presence of atrial fibrillation in patients with asymptomatic and symptomatic left ventricular systolic dysfunction is associated with an increased risk for all-cause mortality, largely explained by an increased risk for pump-failure death. These data suggest that atrial fibrillation is associated with progression of left ventricular systolic dysfunction. PMID- 9741515 TI - Quantitative investigation of cardiomyocyte hypertrophy and myocardial fibrosis over 6 years after cardiac transplantation. AB - OBJECTIVES: This study was performed to determine the degree and time course over 6 years of cardiomyocyte hypertrophy and myocardial fibrosis of the cardiac allograft in transplanted patients. BACKGROUND: Diastolic dysfunction and to a certain extent systolic dysfunction are common cardiac findings after heart transplantation. The development of posttransplant cardiomyocyte hypertrophy and myocardial fibrosis likely contributes to these derangements. METHODS: Cardiomyocyte diameter and percent fibrosis were determined in serial endomyocardial biopsy specimens obtained from 1 month up to 6 years following heart transplantation in 50 patients. Endomyocardial biopsy specimens from 40 patients with primary dilated cardiomyopathy and 11 normal subjects were similarly analyzed for control data. Analyses were performed in a blinded format using a validated computerized image analysis system (Optimas 5.2). RESULTS: Early (1 month) cardiomyocyte enlargement decreased to the smallest diameter 6 months posttransplant, but thereafter progressively increased by 10% to 20% over the subsequent 5- to 6-year period. Although not statistically established, principal stimuli may include a discrepancy in body size (recipient > donor), coronary allograft vasculopathy and posttransplant systemic hypertension. Percent myocardial fibrosis rose early (1 to 2 months) posttransplant and thereafter remained at the same modest level of severity. CONCLUSIONS: Cardiomyocyte diameter of the transplanted heart gradually increases over time, while percent myocardial fibrosis rises early and remains in a modestly elevated plateau after 2 months posttransplant. These histostructural changes likely contribute to the hemodynamic and cardiac functional alterations commonly observed posttransplant. PMID- 9741516 TI - Additional benefit of vitamin E supplementation to simvastatin therapy on vasoreactivity of the brachial artery of hypercholesterolemic men. AB - OBJECTIVES: The aim of this study was to determine whether the combination of lipid-lowering therapy and vitamin E supplementation improves peripheral endothelial function and whether it is more effective than lipid-lowering therapy alone. BACKGROUND: Endothelium-dependent vasodilation is impaired in coronary and peripheral arteries of patients with hypercholesterolemia. Coronary endothelial function has been shown to improve under lipid-lowering and antioxidant therapy, but the effect of additive vitamin E supplementation in the brachial artery is unknown. METHODS: Seven patients with hypercholesterolemia (mean+/-SD; age 51+/ 10 yr) were studied. Endothelium-dependent, flow-mediated dilation (FMD) and endothelium-independent nitroglycerin-induced dilation (NMD) were assessed in the brachial artery using high resolution ultrasound 1) at baseline (BL I), 2) after 8 weeks of simvastatin (20 mg) and vitamin E (300 IU) therapy (Comb I), 3) after withdrawal of vitamin E for 4 weeks (Statin), 4) after therapy as in #2 for 4 weeks (Comb II) and 5) after withdrawal of both drugs for 4 weeks (BL II). RESULTS: Combined simvastatin and vitamin E therapy reduced total cholesterol (Comb I vs. BL I: 276+/-22 vs. 190+/-14 mg/dl, p < 0.0001) and low-density lipoprotein (LDL)-C (197+/-22 vs. 106+/-22 mg/dl, p < 0.00001), augmented alpha tocopherol levels normalized to LDL (12.2+/-4.1 vs. 4.9+/-0.9 microg alpha-T/100 mg% LDL-C, p < 0.01) and resulted in significant improvements in FMD (16.4+/-4.7 vs. 4.9+/-2.5%, p < 0.001) as well as NMD (17.9+/-4.3 vs. 11.2+/-2.8%, p < 0.01). The ratio of FMD to NMD (0.92+/-0.17 vs. 0.46+/-0.24%, p < 0.05) also increased under combination therapy, indicating a greater improvement of FMD than that of NMD. After withdrawal of vitamin E, both FMD (Comb I vs. Statin: 16.4+/-4.7 vs. 7.9+/-4.7%, p < 0.01) and NMD (17.9+/-4.3 vs. 10.9+/-4.5%, p < 0.05) decreased significantly such that simvastatin alone only tended to improve FMD and did not change NMD. Results under combination therapy (Comb II vs. BL II) were reproducible. CONCLUSIONS: Combined vitamin E and simvastatin therapy leads to an improvement of FMD and NMD in the brachial artery of patients with hypercholesterolemia. The improvement of FMD is more pronounced after combination therapy than after lipid-lowering therapy alone, similar to previous findings in the coronary circulation. PMID- 9741517 TI - Mechanisms of hemolysis after mitral valve repair: assessment by serial echocardiography. AB - OBJECTIVES: We sought to determine, using serial echocardiography, the hydrodynamic mechanisms involved in the occurrence of hemolysis after mitral valve repair. BACKGROUND: Recently, fluid dynamic simulation models have identified distinct patterns of mitral regurgitant flow disturbances in patients with mitral prosthetic hemolysis that were associated with high shear stress and may therefore produce clinical hemolysis. Rapid acceleration, fragmentation, and collision jets were associated with high shear stress and hemolysis whereas slow deceleration and free jets were not. METHODS: We reviewed serial echocardiographic studies of 13 consecutive patients with hemolytic anemia after mitral valve repair who were referred for mitral reoperation between January 1985 and December 1996 (group 1). Thirteen patients undergoing reoperation for mitral regurgitation after mitral valve repair but without hemolysis served as controls (group 2). RESULTS: The mitral regurgitant jet was central in origin in 12 group 1 patients and 9 group 2 patients (Fisher exact test, p= 0.3). The other patients had para-ring regurgitation. Group 1 patients had collision (n=11), rapid acceleration (n=2) or fragmentation (n=1) jets whereas group 2 patients had slow deceleration (n=11) or free jets (n=2) (Fisher exact test, p < 0.0001). One patient with hemolysis had both collision and rapid acceleration jets. The "culprit" jet could be identified on the postbypass transesophageal echocardiography (TEE) study in only 1 patient at the time of initial mitral repair. Twelve group 1 patients underwent reoperation, with subsequent resolution of hemolysis in all patients. At reoperation, the initial repair was found to be intact in 8 (67%) patients. CONCLUSION: Distinct patterns of flow disturbance associated with high shear stress were identified by color Doppler imaging in patients with hemolysis after mitral valve repair. The majority (92%) of these color flow disturbances were not present during intraoperative postbypass TEE study after initial mitral repair and subsequently developed in the early postoperative period. PMID- 9741518 TI - Entrainment mapping and radiofrequency catheter ablation of ventricular tachycardia in right ventricular dysplasia. AB - OBJECTIVES: The purpose of this study was to determine if entrainment mapping techniques and predictors of successful ablation sites previously tested in coronary artery disease can be applied to ventricular tachycardia (VT) in arrhythmogenic right ventricular dysplasia (ARVD). BACKGROUND: VT in ARVD has not been well characterized. Reentry circuits in areas of abnormal myocardium are the likely cause, but these circuits have not been well defined. METHODS: Mapping of 19 VTs in 5 patients with ARVD was performed. At 58 sites pacing entrained VT and radiofrequency current (RF) was applied to assess acute termination of VT. RESULTS: Sites classified as exits, central/proximal, inner loop, outer loop, remote bystander and adjacent bystander were identified by entrainment criteria. The reentrant circuit sites were clustered predominantly around the tricuspid annulus and in the right ventricular outflow tract (RVOT). RF ablation acutely terminated VT at 13 sites or 22% of the applications. Of the 19 VTs, eight were rendered noninducible and three were modified to a longer cycle length. In 2 patients ablation at a single site abolished two VTs. CONCLUSION: VT in ARVD shows many of the characteristics of VT due to myocardial infarction. Entrainment mapping techniques can be used to characterize reentry circuits in ARVD. The use of entrainment mapping to guide ablation is feasible. PMID- 9741519 TI - Expanding indications for radiofrequency catheter ablation: ventricular tachycardia in association with right ventricular dysplasia? PMID- 9741520 TI - Role of atrial electrophysiology and autonomic nervous system in patients with supraventricular tachycardia and paroxysmal atrial fibrillation. AB - OBJECTIVES: The purposes of this study were to evaluate the atrial electrophysiology and autonomic nervous system in patients who had paroxysmal supraventricular tachycardia (PSVT) associated with paroxysmal atrial fibrillation (PAF). BACKGROUND: PAF frequently appeared in patients with PSVT. However, the critical determinants for the occurrence of PAF were not clear. METHODS: This study population consisted of 50 patients who had PSVT with (n=23) and without (n=27) PAF. Atrial pressure, atrial size, atrial effective refractory periods (AERPs), and AERP dispersion were evaluated during baseline and PSVT, respectively. Twenty-four hour heart rate variability and baroreflex sensitivity (BRS) were also examined. RESULTS: There was greater baseline AERP dispersion in patients with PAF than in those without PAF. The atrial pressure, atrial size, AERPs in the right posterolateral atrium and distal coronary sinus, and AERP dispersion were increased during PSVT as compared with those during baseline. Patients with PAF had greater AERP dispersion than those without PAF during PSVT. The differences of atrial size, right posterolateral AERP, and AERP dispersion between baseline and PSVT were greater in patients with PAF than in those without PAF. BRS, but not heart rate variability, was higher in patients with PAF than in those without PAF. Univariate analysis showed that higher BRS (>4.5 ms/mm Hg, p=0.0002, odds ratio=16.1), AERP dispersion during PSVT (>40 ms, p=0.0008, odds ratio=9.7), and increase of right atrial area during PSVT (>2 cm2, p=0.016, odds ratio=10.7) were significantly correlated with the occurrence of PAF in patients with PSVT. CONCLUSIONS: Disturbed atrial electrophysiology and higher vagal reflex could play important roles in the genesis of PAF in patients with PSVT. PMID- 9741521 TI - Programmed ventricular stimulation for arrhythmia risk prediction in patients with idiopathic dilated cardiomyopathy and nonsustained ventricular tachycardia. AB - OBJECTIVES: This study investigated the role of programmed ventricular stimulation (PVS) for arrhythmia risk prediction in patients with idiopathic dilated cardiomyopathy (IDC) and spontaneous nonsustained ventricular tachycardia (VT). BACKGROUND: Nonsustained VT in patients with IDC has been associated with a high incidence of sudden cardiac death. METHODS: Over the course of 4 years, 34 patients with IDC, a left ventricular (LV) ejection fraction < or = 35%, and spontaneous nonsustained VT underwent PVS. All patients were prospectively followed for 24+/-13 months. RESULTS: Sustained ventricular arrhythmias were induced in 13 patients (38%). Sustained monomorphic VT was induced in three patients (9%), and polymorphic VT or ventricular fibrillation (VF) in another 10 patients (29%). No sustained ventricular arrhythmia could be induced in 21 study patients (62%). Prophylactic implantation of third-generation defibrillators (ICDs) with electrogram storage capability was performed in all 13 patients with inducible sustained VT or VF, and in nine of 21 patients (43%) without inducible sustained VT or VF. There were no significant differences between the additional use of amiodarone, d,I-sotalol, and beta-blocker therapy during follow-up in patients with and without inducible VT or VF. During 24+/-13 months of follow-up, arrhythmic events were observed in nine patients (26%) including sudden cardiac deaths in two patients and ICD shocks for rapid VT or VF in seven patients. Arrhythmic events during follow-up occurred in four of 13 patients with inducible ventricular arrhythmias compared with five of 21 patients without inducible ventricular arrhythmias at PVS (31% vs. 24%, p=NS). CONCLUSION: PVS does not appear to be helpful for arrhythmia risk stratification in patients with IDC, a left ventricular ejection fraction < or =35%, and spontaneous nonsustained VT. Due to the limited number of patients, however, the power of this study is too small to exclude moderately large differences in outcome between patients with IDC with and without inducible VT or VF. PMID- 9741522 TI - Improved left ventricular endocardial border delineation and opacification with OPTISON (FS069), a new echocardiographic contrast agent. Results of a phase III Multicenter Trial. AB - OBJECTIVES: The echocardiographic contrast-enhancing effects and safety profile of ALBUNEX (a suspension of air-filled albumin microspheres) were compared with the new contrast agent OPTISON (formerly FS069: a suspension of albumin microspheres containing the gas perfluoropropane) in 203 patients with inadequate noncontrast echocardiograms. BACKGROUND: The efficacy of ALBUNEX has been limited by its short duration of action. By using perfluoropropane instead of air within the microsphere, its duration of action is increased. METHODS: Each patient received ALBUNEX (0.8 and 0.22 mL/kg) and OPTISON (0.2, 0.5, 3.0, and 5.0 mL) on separate days a minimum of 48 hours apart. Echocardiograms were evaluated for increase in left ventricular (LV) endocardial border length, degree of LV opacification, number of LV endocardial border segments visualized, conversion from a nondiagnostic to a diagnostic echocardiogram, and duration of contrast enhancement. A thorough safety evaluation was conducted. RESULTS: Compared with ALBUNEX, OPTISON more significantly improved every measure of contrast enhancement. OPTISON increased well-visualized LV endocardial border length by 6.0+/-5.1, 6.9+/-5.4, 7.5+/-4.7, and 7.6+/-4.8 cm, respectively, for each of the four doses, compared with only 2.2+/-4.5 and 3.4+/-4.6 cm, respectively, for the two ALBUNEX doses (p < 0.001). 100% LV opacification was achieved in 61%, 73%, 87%, and 87% of the patients with the four doses of OPTISON, but in only 16% and 36% of the patients with the two ALBUNEX doses (p < 0.001). Conversion of nondiagnostic to diagnostic echocardiograms with contrast occurred in 74% of patients with the optimal dose of OPTISON (3.0 mL) compared with only 26% with the optimal dose of ALBUNEX (0.22 mL/kg) (p < 0.001). The duration of contrast effect was also significantly greater with OPTISON than with ALBUNEX. In a subset of patients with potentially poor transpulmonary transit of contrast (patients with chronic lung disease or dilated cardiomyopathy), OPTISON more significantly improved the same measures of contrast enhancement compared with ALBUNEX and did so to the same extent as in the overall population. Side effects were similar and transient with the two agents. CONCLUSION: OPTISON appears to be a safe, well tolerated echocardiographic contrast agent that is superior to ALBUNEX. PMID- 9741523 TI - Mortality in potential arterial switch candidates with transposition of the great arteries. AB - OBJECTIVES: We reviewed the factors contributing to or causing death before surgery in neonates with d-transposition of the great arteries (TGA) despite anatomy suitable for the arterial switch operation (ASO) to develop strategies to minimize preoperative attrition. BACKGROUND: Currently the ASO for neonates with TGA carries a low operative mortality. However, there is a paucity of information regarding the patients who die before the ASO. Strategies to ensure survival to operation are of importance to improve overall outcome. METHODS: We reviewed all neonates with TGA and patent forearm ovale (PFO) < or = 2 mm, a birthweight <2 kg, or who died before surgery, between 1988 and 1996. RESULTS: We identified 12 out of 295 neonates with TGA (4.1%) with anatomy suitable for the ASO who died prior to surgery. All had TGA/intact ventricular septum (IVS) and presented with a severely restrictive PFO. In 11 of 12 cases the cause of death was attributed to the sequelae of profound hypoxemia from inadequate mixing. Contributing factors were prematurity, 41.7%; severe respiratory distress syndrome, 25%; and persistent pulmonary hypertension of the newborn (PPHN), 16.7%. All patients received prostaglandin E1 (PGE1) infusion. Urgent balloon atrial sepstostomy (BAS) was performed in 66.7% with improved oxygenation. No cases were diagnosed prenatally. In contrast, all patients with a PFO < or = 2 mm who survived to ASO had a significantly better response to PGE1 infusion (p=0.03) than nonsurvivors. The ASO was accomplished without mortality in four of nine with a weight <2 kg. CONCLUSIONS: Of those neonates admitted with TGA, 4.1% died before surgery. Eleven of 12 (3.7%) died due to consequences of inadequate interatrial mixing despite PGE1 infusion. Earlier diagnosis and BAS are critically important in determining survival. Early ASO may improve survival in patients weighing <2 kg. Prenatal diagnosis with delivery in a high-risk obstetrical unit with facilities for immediate BAS and supportive therapy for pulmonary hypertension and ventricular failure may be necessary to salvage this group of patients. PMID- 9741524 TI - Long-term outcome after the mustard repair for simple transposition of the great arteries. 28-year follow-up. AB - OBJECTIVES: This study examines the late outcome in patients with simple transposition of the great arteries (TGA) after a Mustard operation. BACKGROUND: Continuing medical follow-up for patients after the Mustard procedure, now extending to three decades, is required. The quality of life of adult survivors has not been well documented. METHODS: Survival and quality of life among 113 hospital survivors of the Mustard operation performed for simple TGA between 1964 and 1982 were assessed by medical review and a lifestyle questionnaire. The incidence of right ventricular failure and echocardiographic right ventricular dysfunction (RVD) were determined. A measure of lifestyle, the ability index, was determined. RESULTS: Actuarial survival was 90%, 80%, and 80% at 10, 20, and 28 years, respectively, with 76% of survivors being New York Heart Association class 1. Sudden death, with an incidence of 7% without identifiable risk factors, was the most common cause of late demise. RVD was identified in 18% of patients who had echocardiography, but there was right ventricular failure in only two patients. Seventy-five percent of current survivors lead a normal life, 20% have some symptoms or lifestyle modification, and 5% are unable to work. CONCLUSIONS: The survival of patients to 28 years with the Mustard repair has been good. Late sudden death is the most worrisome feature. There is a 97% freedom from right ventricular failure to date. The quality of life of late survivors is good, most achieving a normal level of education and employment. PMID- 9741525 TI - Effect of veno-venous ultrafiltration on myocardial performance immediately after cardiac surgery in children. A prospective randomized study. AB - OBJECTIVES: This study sought to evaluate the effects of veno-venous ultrafiltration on myocardial contractility in children undergoing cardiopulmonary bypass (CPB) for repair of congenital heart defects. BACKGROUND: Ultrafiltration (UF) is currently used to diminish postoperative fluid accumulation following CPB in children. Previous reports indicate improvement in hemodynamics immediately after UF, but the mechanism of its action is unknown. METHODS: Twenty-three patients (ages 2 months to 9.1 years; 13 males, 10 females) underwent UF for 10 min after CPB. Twelve patients underwent UF immediately after CPB (Group A). They were studied: (1) before and (2) after CPB, (3) after UF, and (4) 10 min after UF. Eleven patients underwent UF 10 min after CPB (Group B). They were studied: (1) before and (2) after CPB, (3) after a 10-min delay before UF, and (4) after UF. Contractility was determined by the difference in the observed and predicted velocity of circumferential fiber shortening for the measured wall stress, using transesophageal echocardiography. Left ventricular wall thickness was also measured. RESULTS: There was significant improvement in contractility after UF in both groups (mean+/-SD, Group A: -0.28+/-0.13 to 0.01+/-0.21 circ/s, p < 0.05; Group B: -0.26+/-0.16 to -0.11+/-0.17 circ/s, p < 0.05). Myocardial thickness to cavity dimension decreased in both groups following UF (Group A: 0.19+/-0.04 to 0.14+/-0.03, p < 0.05; Group B: 0.18+/-0.04 to 0.14+/-0.03, p < 0.05). CONCLUSIONS: UF improves hemodynamics by improving contractility and possibly by reducing myocardial edema in children following cardiac surgery. Enhanced patient outcome after ultrafiltration may in part be due to these changes. PMID- 9741526 TI - Supraventricular tachycardia in patients with right atrial isomerism. AB - OBJECTIVES: To clarify the prevalence and mechanism of supraventricular tachycardia in patients with right atrial isomerism. BACKGROUND: Paired SA and dual atrioventricular (AV) nodes have been described in patients with right atrial isomerism. However, the clinical significance remains unclear. METHODS: From 1987 to 1996, a total of 101 patients (61 male, 40 female) and four fetuses were identified with right atrial isomerism. The diagnosis of supraventricular tachycardia exclude the tachycardia with prolonged QRS duration or AV dissociation, and primary atrial tachycardia. RESULTS: The median follow-up duration was 38 months (range 0.2-270 months). Supraventricular tachycardia was documented in 25 patients (24.8%) and one fetus (25%) (onset age ranged from prenatal to 14 years old; median 4 years old). Actuarial Kaplan-Meier analysis revealed that the probability of being free from tachycardia was 67% and 50% at 6 and 10 years of age, respectively. These tachycardias could be converted by vagal maneuvers in one, verapamil in seven, propranolol in four, digoxin in two, procainamide in one, and rapid pacing in five. Spontaneous conversion was noted in six (including the fetus). Seven cases had received electrophysiological studies. Reciprocating AV tachycardia could be induced in five and echo beats in one. The tachycardia in three patients was documented as incorporating a posterior AV node (antegrade) and an anterior or a lateral AV node (retrograde). Two of them received radiofrequency ablation. Successful ablation in both was obtained by delivering energy during tachycardia, aimed at the earliest retrograde atrial activity and accompanied by junctional ectopic rhythm. The patient with echo beats developed tachycardia soon after operation. CONCLUSIONS: Supraventricular tachycardia is common in patients with right atrial isomerism and can occur during the prenatal stage. Drugs to slow conduction through the AV node may help to terminate the tachycardia. Radiofrequency ablation is a safe and effective treatment alternative to eliminate tachycardia. PMID- 9741527 TI - Intramural delivery of a specific tyrosine kinase inhibitor with biodegradable stent suppresses the restenotic changes of the coronary artery in pigs in vivo. AB - OBJECTIVES: This study was designed to examine whether or not intramural delivery of ST638 (a specific tyrosine kinase inhibitor) with biodegradable stent can suppress the restenotic changes of the coronary artery in vivo. BACKGROUND: Clinical and animal studies demonstrated that restenosis after coronary intervention results from a combined effect of neointimal formation and geometric remodeling (decrease in total cross-sectional area). Thus, the most effective strategy to prevent the restenosis appears to inhibit both the neointimal formation and geometric remodeling by antiproliferative agent and stent, respectively. We have previously shown that ST638 markedly suppresses the restenotic changes of the porcine coronary artery when applied from the adventitial site. METHODS: A poly-L-lactic acid biodegradable stent was coated with either ST638 (0.8 mg) or equimolar of its inactive metabolite, ST494. A pair of these stents were implanted alternatively in the left anterior descending or circumflex coronary artery in pigs (n=6). Three weeks after the procedure, coronary stenosis was assessed by angiography followed by histological examination. RESULTS: Coronary stenosis was significantly less at the ST638 stent site than at the ST494 stent site (47+/-5% vs. 25+/-4%, p < 0.01). Histological examination also showed that the extent of neointimal formation and that of geometric remodeling were significantly less at the ST638 stent site than at the ST494 stent site (p < 0.05). CONCLUSIONS: These results indicate that intramural delivery of a specific tyrosine kinase inhibitor with biodegradable stent overcomes the proliferative stimuli caused by balloon injury, the stent itself, and the drug coating on the stent, resulting in the suppression of the restenotic changes of the coronary artery in vivo. This strategy might also be useful in the clinical setting in humans. PMID- 9741528 TI - Microvascular injury in reperfused infarcted myocardium: noninvasive assessment with contrast-enhanced echoplanar magnetic resonance imaging. AB - OBJECTIVES: The purpose of this study was to measure the accumulation of labeled albumin and to visualize its distribution pattern in reperfused infarcted myocardium as a function of time between onset of reperfusion and administration of the tracer. BACKGROUND: Myocardial microvascular injury leads to leakage of albumin from the intravascular space. Quantitative measurements of GdDTPA-albumin with inversion recovery echoplanar imaging (IR-EPI) may allow noninvasive monitoring of microvascular injury. METHODS: After 1 h of coronary artery occlusion, 56 rats were injected with GdDTPA-albumin or 123I-GdDTPA-albumin either immediately before reperfusion or 1/2, 1 or 24 h after reperfusion. GdDTPA albumin in blood, normal myocardium and reperfused infarction was dynamically measured with IR-EPI during 1 h postinjection (PI). Autoradiograms were obtained at 15 min PI. Accumulation of labeled albumin in myocardium was expressed as the ratio of myocardial to blood content. RESULTS: In normal myocardium, the ratio of changes of relaxation rate-ratio (deltaR1-ratio) was 0.12+/-0.01 and did not change over 1 h. In reperfused infarction, however, the deltaR1-ratio increased after administration. Animals given GdDTPA-albumin before reperfusion exhibited fastest accumulation (deltaR1-ratio 15 min PI: 0.56+/-0.03) and essentially homogeneous distribution. The accumulation was slower when administered at 1/2, 1 and 24 h after reperfusion (deltaR1-ratios 15 min PI: 0.39+/-0.03; 0.31+/-0.04; 0.16+/-0.01; p < 0.001 compared to administration before reperfusion). Moreover, the tracer accumulated predominantly in the periphery of the injury zone. CONCLUSIONS: Amount and distribution pattern of labeled albumin in reperfused infarction are modulated by duration of reperfusion. The accumulation of GdDTPA albumin can be quantified by IR-EPI. Thus, IR-EPI may be useful to noninvasively monitor myocardial microvascular injury in reperfused infarction. PMID- 9741529 TI - Validation of the in vivo intravascular ultrasound measurement of in-stent neointimal hyperplasia volumes. AB - OBJECTIVES: This study was undertaken to validate the in vivo intravascular ultrasound (IVUS) measurement of in-stent neointimal hyperplasia (IH) volumes. BACKGROUND: Because stents reduce restenosis compared to balloon angioplasty, stent use has increased significantly. As a result, in-stent restenosis is now an important clinical problem. Serial IVUS studies have shown that in-stent restenosis is secondary to intimal hyperplasia. To evaluate strategies to reduce in-stent restenosis, accurate measurement of in-stent neointimal tissue is important. METHODS: Using a porcine coronary artery model of in-stent restenosis, single Palmaz-Schatz stents were implanted into 16 animals with a stent:artery ratio of 1.3:1. Intravascular ultrasound imaging was performed at 1 month, immediately prior to animal sacrifice. In vivo IVUS and ex vivo histomorphometric measurements included stent, lumen and IH areas; IH volumes were calculated with Simpson's rule. RESULTS: Intravascular ultrasound measurements of IH (30.4+/-11.0 mm3) volumes correlated strongly with histomorphometric measurements (26.7+/-8.5 mm3, r=0.965, p < 0.0001). The difference between the IVUS and the histomorphometric measurements of IVUS volume was 4.1+/-2.7 mm3 or 15.8+/-11% (standard error of the estimate=0.7). Both histomorphometry and IVUS showed that IH was concentric and uniformly distributed over the length of the stent. Intravascular ultrasound detected neointimal thickening of < or =0.2 mm in 5 of 16 stents. Sample size calculations based on the IVUS measurement of IH volumes showed that 12 stented lesions/arm would be required to show a 50% reduction in IVUS-measured IH volume and 44 stented lesions/arm would be required to show a 25% reduction in IH volume. CONCLUSION: In vivo IVUS measurement of IH volumes correlated strongly with ex vivo histomorphometry. Using volumetric IVUS end points, small sample sizes should be necessary to demonstrate effectiveness of strategies to reduce in-stent restenosis. PMID- 9741530 TI - Effects of angiotensin II on expression of the gap junction channel protein connexin43 in neonatal rat ventricular myocytes. AB - OBJECTIVES: To elucidate signal transduction pathways regulating expression of myocardial gap junction channel proteins (connexins) and to determine whether mediators of cardiac hypertrophy might promote remodeling of gap junctions, we characterized the effects of angiotensin II on expression of the major cardiac gap junction protein connexin43 (Cx43) in cultured neonatal rat ventricular myocytes. BACKGROUND: Remodeling of the distribution of myocardial gap junctions appears to be an important feature of anatomic substrates of ventricular arrhythmias in patients with heart disease. Remodeling of intercellular connections may be initiated by changes in connexin expression caused by chemical mediators of the hypertrophic response. METHODS: Cultures were exposed to 0.1 micromol/liter angiotensin II for 6 or 24 h, and Cx43 expression was characterized by immunoblotting, confocal microscopy and electron microscopy. RESULTS: Immunoblot analysis revealed a twofold increase in Cx43 content in cells treated for 24 h with angiotensin II (n=4, p < 0.05). This response was inhibited by the presence of 1.0 micromol/liter losartan, an AT1-receptor blocker. Confocal and electron microscopy demonstrated enhanced Cx43 immunoreactivity and increases in the number and size of gap junction profiles in cells exposed to angiotensin II for 24 h. These effects were also blocked by losartan. Immunoprecipitation of Cx43 from cells metabolically labeled with [35S]methionine demonstrated 2.4- and 2.9-fold increases in Cx43 radioactivity after 6 and 24 h exposure to angiotensin II, respectively (p < 0.03 at each time point). CONCLUSIONS: Angiotensin II up regulates gap junctions in cultured neonatal rat ventricular myocytes by increasing Cx43 synthesis. Signal transduction pathways activated by angiotensin II under pathophysiologic conditions could initiate remodeling of conduction pathways, leading to the development of anatomic substrates of arrhythmias. PMID- 9741531 TI - Denopamine, a beta1-adrenergic agonist, prolongs survival in a murine model of congestive heart failure induced by viral myocarditis: suppression of tumor necrosis factor-alpha production in the heart. AB - OBJECTIVES: This study was designed to examine the effects of denopamine, a selective beta1-adrenergic agonist, in a murine model of congestive heart failure (CHF) due to viral myocarditis. BACKGROUND: Positive inotropic agents are used to treat severe heart failure due to myocarditis. However, sympathomimetic agents have not been found beneficial in animal models of myocarditis. METHODS: In vitro: The effects of denopamine on lipopolysaccharide-induced tumor necrosis factor-alpha (TNF-alpha) production was studied in murine spleen cells. In vivo: Four-week-old DBA/2 mice were inoculated with the encephalomyocarditis virus (day 0). Denopamine (14 micromol/kg), denopamine (14 micromol/kg) with a selective beta1-blocker metoprolol (42 micromol/kg), or denopamine (14 micromol/kg) with metoprolol (84 micromol/kg) was given daily, and control mice received the vehicle only. Survival and myocardial histology on day 14 and TNF-alpha levels in the heart on day 6 were examined. RESULTS: In the in vitro study, TNF-alpha levels in treated cells were significantly lower than in controls (p < 0.05). In the in vivo study treatment with denopamine significantly improved the survival of the animals (14 of 25 (56%) treated, vs 5 of 25 (20%) control mice), attenuated myocardial lesions, and suppressed TNF-alpha production (66.5+/-7.5 pg/mg of heart in treated mice vs 113.5+/-15.1 pg/mg of heart in control mice, mean+/-SE). There was a strong linear relationship between mortality and TNF alpha levels (r=0.98, n=4, p < 0.05). These in vitro and in vivo effects of denopamine were significantly inhibited by metoprolol. CONCLUSIONS: These results suggest that denopamine may exert its beneficial effects, in part, by suppressing the production of TNF-alpha via beta1-adrenoceptors. PMID- 9741532 TI - Effects of critical coronary stenosis on global systolic left ventricular function quantified by pressure-volume relations during dobutamine stress in the canine heart. AB - OBJECTIVES: In this study we quantified the effects of a critical coronary stenosis on global systolic function using pressure-volume relations at baseline and during incremental dobutamine stress. BACKGROUND: The effects of coronary stenosis have previously been analyzed mainly in terms of regional (dys)function. Global hemodynamics are generally considered normal until coronary flow is substantially reduced. However, pressure-volume analysis might reveal mechanisms not fully exposed by potentially load-dependent single-beat parameters. Moreover, no systematic analysis by pressure-volume relations of the effects of dobutamine over a wide dose range has previously been presented. METHODS: In 14 dogs left ventricular volume and pressure were measured by conductance and micromanometer catheters, and left circumflex coronary flow by Doppler probes. Measurements in control and with left circumflex stenosis were performed at baseline and at five levels of dobutamine (2.5 to 20 microg/kg/min). The end-systolic pressure-volume relation (ESPVR) dP/dtMAX vs. end-diastolic volume (dP/dtMAX - V(ED)) and the relation between stroke work and end-diastolic volume (preload recruitable stroke work [PRSW]) were derived from data obtained during gradual caval occlusion. RESULTS: In control, dobutamine gradually increased heart rate up to 20 microg/kg/min, the inotropic effect blunted at 15 microg/kg/min. With stenosis, the chronotropic effect was similar, however, contractile state was optimal at approximately 10 microg/kg/min and tended to go down at higher levels. At baseline, the positions of ESPVR and PRSW, but not of dP/dtMAX - V(ED), showed a significant decrease in function with stenosis. No differences between control and stenosis were present at 2.5 microg/kg/min; the differences were largest at 15 microg/kg/min. CONCLUSIONS: Pressure-volume relations and incremental dobutamine may be used to quantify the effects of critical coronary stenosis. The positions of these relations are more consistent and more useful indices than the slopes. The positions of the ESPVR and PRSW show a reduced systolic function at baseline, normalization at 2.5 microg/kg/min and a consistent significant difference between control and stenosis at dobutamine levels of 5 microg/kg/min and higher. PMID- 9741533 TI - The ACC professional life survey: career decisions of women and men in cardiology. A report of the Committee on Women in Cardiology. American College of Cardiology. AB - OBJECTIVES: This survey was conducted to learn how the career decisions of women and men in cardiology influenced their professional and personal lives. BACKGROUND: Women represent only 5% of practicing adult cardiologists and 10% of trainees. Yet, women and men now enter medical school at nearly equal numbers. The factors that contribute to career satisfaction in cardiology should be identified to permit the development of future strategies to ensure that the best possible candidates are attracted to the profession. METHODS: A questionnaire developed by the Ad Hoc Committee on Women in Cardiology of the American College of Cardiology (ACC) was mailed in March 1996 to all 964 female ACC members and an age-matched sample of 1,199 male members who had completed cardiovascular training. RESULTS: Women were more likely to describe their primary or secondary role as a clinical/noninvasive than invasive cardiologist (p < 0.0001 women vs. men). Men and women both reported a high level of satisfaction with family life, but women were less satisfied with their work as cardiologists (88% vs. 92%, p < 0.01) and with their level of financial compensation. Compared with men, women expressed less overall satisfaction (69% vs. 84%) and more dissatisfaction with their ability to achieve professional goals (21% vs. 9%). These differences were most pronounced for women in academic practice. Women reported greater family responsibilities, which may limit their opportunities for career advancement. Women were more likely to alter training or practice focus to avoid radiation. A majority of women (71%) reported gender discrimination, whereas only 21% of men reported any discrimination, largely due to race, religion or foreign origin. CONCLUSIONS: Women cardiologists report overall lower satisfaction with work and advancement, particularly within academic practice. They report more discrimination, more concerns about radiation and more limitations due to family responsibilities, which may ultimately explain the low percentage of women in cardiology. Attention to these issues may result in programs to improve professional satisfaction and attract the best candidates into cardiology in the future. PMID- 9741534 TI - The Olympics, NBA Playoffs, U.S. Open, the World Cup, and Wimbledon--points to ponder. PMID- 9741535 TI - ACC expert consensus document. Present use of bedside right heart catheterization in patients with cardiac disease. American College of Cardiology. PMID- 9741536 TI - Genetic events underlying morphological complexity of gastric carcinoma. AB - Cancer is a genetic disorder in which gene alterations are selected to provide growth advantage by oncogene activation and/or tumor suppressor gene inactivation. Even marked intra-tumor variation in the histologic pattern, which is common in gastric carcinoma, is considered a result of distinct oncogenic pathways coexisting together. The present review describes that most gastric carcinomas arise through two distinct genetic pathways: microsatellite instability targeting the mononucleotide tracts within coding regions of cancer related genes and chromosomal deletion involving tumor suppressor genes. With regard to malignant phenotypes, microsatellite instability is associated with the intestinal histological type and chromosomal deletion is correlated with the growth pattern of gastric carcinoma. Moreover, the genetic instability would in turn lead to an increase in alterations of cancer-related genes. The corresponding cells gradually manifest diverse neoplastic properties, thus bringing about consecutive subclonal evolution of more malignant cells. We now have some dues leading to the characterization of phenotypic complexity of gastric carcinoma based on gene-inactivation mechanisms. PMID- 9741537 TI - Regeneration of graft liver in adult-to-adult living donor liver transplantation using a left lobe graft. AB - Graft size-matching is one of the critical concerns in adult-to-adult living donor liver transplantation (ATALDLT). In this study, we evaluated regeneration of a small-for-size graft less than 50% of the standard liver volume (SLV). We reviewed nine patients of united network of organ sharing (UNOS) status 2 or 3 who had undergone ATALDLT with a left lobe graft. For the comparison of liver regeneration, 20 hepatectomized patients for biliary malignancy were selected as non-transplant control group. In the ATALDLT group, graft size ranged from 30 to 49% of the SLV of recipients and their regeneration rates were 158%, 182%, 200% and 185% after 1,2, 3 and 4 weeks following transplantation, respectively. In the control group, preoperative volume of left lobe to whole liver volume ranged between 40 and 54% and their regeneration rates were 145%, 156%, 163% and 177% after 1,2, 3 and 4 weeks following extended right lobectomy, respectively. There was no statistical difference in regeneration rates between two groups. In the ATALDLT group, serum aspartate aminotransferase showed the median peak level of 198 IU/L on the first postoperative day and it was normalized within one week. Recovery of bilirubin clearance lagged behind rapid volume regeneration by about one week. Two patients died of sepsis. We postulate that the regenerative power of small-for-size grafts from living donors is preserved, although time-lag between volume regeneration and metabolic capability occurs in small-for-size grafts, when the initial graft volume meets metabolic demands during the early postoperative days. PMID- 9741538 TI - Phylogenetically interrelated ETS genes, ETV1, ERM and E1A-F locate on different chromosomes. AB - ETV1, ERM and E1A-F are members of the multigene ETS domain containing a class of transcription factors, first identified in the genome of the avian retrovirus E26. Based upon extensive homology between these genes within their ETS domain (96% identity each other), these three genes comprise a distinct sub-family of ETS genes as a human PEA3 sub-family. By analyzing somatic cell hybrid segregating human chromosomes, the genes encoding ETV1, ERM and E1A-F were localized to human chromosome 7, 3 and 17, respectively. Fluorescence in situ hybridization confirmed these assignments and allowed mapping of the genes to 7p22 (ETV1), 3q29 (ERM) and 17q12 (E1A-F). These results suggest the ancestral PEA3 gene may have duplicated first, then dispersed to other chromosomal locations. PMID- 9741539 TI - Immunohistochemical and SSCP analysis of p53 in malignant lymphomas. AB - We immunohistochemically investigated Epstein-Barr virus (EBV)-positive and negative 31 malignant lymphomas (MLs) for p53 protein using a monoclonal antibody which is expressed on a wild type and mutant human p53 protein. We evaluated the presence of mutations in exons 5 to 8 of the p53 gene using single-strand confirmation polymorphism analysis. Overexpression of p53 was detected in 13 out of 31 cases (41.9%) of MLs. However, we have documented the presence of structural alterations of the p53 gene in six cases of MLs. The presence of EBV infection in MLs was statistically unrelated to p53 protein overexpression. Excellent correlation was found between p53 immunoreactivity and histologic types of MLs. Even though the reason for discrepancy between p53 gene mutation and p53 protein overexpression remains unclear, p53 protein overexpression may be involved in the process of malignant transformation regardless of EBV infection in MLs. PMID- 9741540 TI - Expression of cyclin dependent kinase inhibitor p21WAF1 alone and in combination with p27KIP1 shows prognostic value in gastric carcinoma. AB - Evidence suggests that multiple molecular events, including alteration of cell cycle regulators are involved in the development and progression of gastric carcinoma. Recently, it has been reported that the expression of p21 and p27, integrating the effects of one or more cell cycle regulators, consequently, acting as a single indicator of several possible cell cycle gene alterations is associated with tumor suppression. In the present study, we studied the immunohistochemical expression of p21 and p27 in gastrectomy specimens from 84 patients with gastric adenocarcinoma, and analysed its correlation to clinicopathologic data, including patients survival. Loss of p21 and p27 expression was noted in 45 (53.6%) and 44 (52.4%) of the 84 gastric carcinoma tissues, respectively. The expression of p21 was significantly correlated with histological type (p= 0.005), recurrence (p=0.002) and death (p=0.002) after surgery, and p27 expression (p=0.001). Kaplan-Meier survival plots showed p21 negative group (p= 0.0014) or both p21 and p27 negative group (p=0.0048) was significantly poorer in overall survival than both p21 and p27 positive or one of both positive group. Our results suggest that the status of p21 and p27 expression in immunohistochemical stain may be a useful prognostic marker of gastric carcinoma. PMID- 9741541 TI - Localization of hepatitis B virus DNA in hepatocellular carcinoma by polymerase chain reaction in situ hybridization. AB - The polymerase chain reaction in situ hybridization (PCR-ISH) is a new technique that combines the sensitivity of PCR with the localizing ability of ISH. To investigate the expression pattern of hepatitis B virus (HBV) in the tissue of hepatocellular carcinoma (HCC), we detected HBV-DNA with PCR-ISH in paraffin embedded tumor and corresponding non-tumor tissues from 11 HCC patients. HBV-DNA was detected in 4 of 11 tumor tissues and in 7 of 10 non-tumor tissues. In tumor tissues, positive signals were scattered in the tissue with occasional clustering, and were found mainly in the cytoplasm of HCC cells rather than in the nucleus. In non-tumor tissues, the number of positive signals was higher than in tumor tissues and they were found in regenerating nodules with differing patterns and intensities. When we compared the detection rate of PCR-ISH with nested PCR among 10 tissue samples, HBV-DNA was detected in 5 tissue samples by PCR-ISH, but the S gene was detected in 10, precore gene in 9 and X gene in 8 by nested PCR. The findings suggest that PCR-ISH is a sensitive technique for localizing HBV in tissue sections and that the low level of HBV replication persists in HCC cells. PMID- 9741542 TI - HSP70 and ER expression in cervical intraepithelial neoplasia and cervical cancer. AB - Heat shock protein (HSP) is thought to play important roles in the cell cycle and various process of carcinogenesis. This study was performed to evaluate the expression of heat shock protein (HSP70) and estrogen receptor (ER) and Ki-67 and to assess relationship between them in cervical squamous cell neoplasia. The materials were 50 cervical squamous cell lesions, consisted of 30 cervical intraepithelial neoplasia (CIN) (6 moderate dysplasia, 11 severe dysplasia, 13 carcinoma in situ), and 20 invasive squamous cell carcinoma (ISCC) cases. These specimens were immunohistochemically stained for HSP70, ER and Ki-67. The score of HSP70 was significantly higher in ISCC than CIN. Expression rate of the ER was not significantly higher in CIN than in ISCC. Ki-67 labelling index was significantly higher in the ISCC and high HSP70 positive staining group. These results suggested that HSP70 may play an important role in tumor cell proliferation and is related with ISCC than CIN, but ER may be not related with tumor cell proliferation and differentiation. HSP70 may be a useful prognostic factor in cervical dysplasia and cancer. PMID- 9741543 TI - Metabolic acidosis and urinary acidification defect during the course of hemorrhagic fever with renal syndrome. AB - To evaluate urinary acidification defect and its contribution to metabolic acidosis (MA) during hemorrhagic fever with renal syndrome (HFRS), we serially analyzed acid-base balance and urinary acidification indices in 10 HFRS patients. Data of the patients were compared with those of 8 normal volunteers (NC). MA was observed in 6 of 8 patients in the oliguric phase, 5 of 7 in the early diuretic phase, 8 of 10 in the late diuretic phase and 2 of 9 in the convalescent phase. HFRS patients with MA had a higher plasma anion gap in the oliguric and early diuretic phases than NC and a higher plasma Cl/Na ratio in the late diuretic phase than NC. As compared with acid-loaded NC, HFRS patients had a higher urine pH in the oliguric, early diuretic and late diuretic phases, a higher urine anion gap (UAG) in the oliguric and early diuretic phases and a lower urinary NH4+ excretory rate in the oliguric, early diuretic and late diuretic phases. Overt distal acidification defect was observed in 6 of 8 patients in the oliguric phase, 3 of 7 in the early diuretic phase, 5 of 10 in the late diuretic phase and none of 9 in the convalescent phase. None of the convalescent patients had latent acidification defect. In conclusion, urinary acidification defect is marked in the oliguric and diuretic phases of severe HFRS and may play a role in the development of a high anion gap (AG) metabolic acidosis in the earlier phase and hyperchloremic MA in the later phase, but rapidly recovers in the convalescent phase. PMID- 9741544 TI - Detection of t(11;22)(q24;q12) translocation of Ewing's sarcoma in paraffin embedded tissue by nested reverse transcription-polymerase chain reaction. AB - Ewing's sarcoma is a poorly characterized malignant tumor with a relatively uniform histologic appearance, made up of densely packed small cells with round to oval nuclei, without distinct cell borders and without any structural differentiation. Often the diagnosis has to be made by exclusion. Recently, it has been made possible to identify characteristic chromosomal rearrangements associated with certain solid tumors. More than 85% of Ewing's sarcoma and peripheral neuroectodermal tumor present a specific t(11;22)(q24;q12) balanced translocation, resulting in the production of a novel chimerical EWS/FLI-1 message. Using oligonucleotide primers derived from EWS and FLI-1 complementary DNAs, we were able to use reverse transcription-polymerase chain reaction (RT PCR) as a diagnostic tool. The described nested RT-PCR method as another supportive diagnostic method enables pathologists to differentiate small blue cell tumors not only to make correct diagnosis but also to investigate retrospective archival tumor samples, using formalin fixed paraffin embedded tissue as a source of RNA. PMID- 9741545 TI - Anticardiolipin and anti-beta2-glycoprotein I antibodies in Behcet's disease. AB - To investigate prevalence of anticardiolipin antibodies (aCL) in patients with Behcet's disease (BD) and to determine whether they are related to anti-beta2 glycoprotein I antibodies (aGPI), we measured aCL and aGPI in 47 patients of BD and 14 patients of systemic lupus erythematosus (SLE). The levels of aCL and aGPI were determined by conventional enzyme immunoassay for both IgG and IgM classes. Twelve (25.5%) patients with BD were positive for IgG or IgM aCL and no patient was positive for aGPI. Eleven (78.6%) patients with SLE were also positive for aCL and among them, 8 (72.7%) patients were positive for aGPI. Positive IgG aCL patients with BD showed lower level of IgG aCL than those with SLE (15.7+/-7.3 vs 34.1+/-16.0 GPL, p<0.05). There was no relation between the presence of aCL in BD and either dinical activity or clinical features. In the patients with BD, aCL are found but it would not be associated with aGPI as they are in patients with SLE. In patients with BD, aCL seem to be authentic aCL unlike those in patients with SLE and may not be related with vascular complications in BD. PMID- 9741546 TI - Cytokinetic pattern in the thoracic spinal cord of chick embryos (incubation day 5-13) using PCNA staining and TUNEL method. AB - For the cytokinetic studies using spinal cords of chick embryos, chronological patterns of cell proliferation and programmed cell death (apoptosis) should be known. Information in the early stages of chick embryos is available while data on later stages are seldom available. To investigate the chronological patterns of cell proliferation and apoptosis in the thoracic spinal cord of normal chick embryos on incubation day 5, 6, 8, 10 and 13 (Hamburger and Hamilton stage 26 40), proliferating cell nuclear antigen (PCNA) staining and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) method were used. Cell proliferation was active at the germinal layer on days 5 and 6. It markedly declined on day 8 and became negligible on day 13. TUNEL-positive cells were mainly found in the germinal layer, the ventrolateral part of the mantle layer and the dorsal root ganglion. Compared to PCNA-positive cells, TUNEL positive cells were sparse, especially after day 10, when only a few positive cells were scattered. These results will be used as a control data for the studies such as an experimental research for neural tube defects. PMID- 9741547 TI - Indoleamine-accumulating cell death and endogenous glial cell reaction induced by 5,7-dihydroxytryptamine in the cat retina. AB - We investigated the patterns of degenerative changes of indoleamine-accumulating cells (IACs) induced by 5,7-dihydroxytryptamine (5,7-DHT, 100 microg), and the glial reaction to the neurodegenerative changes of IACs in the cat retina by using light-and electron-microscopy. The neurons accumulating 5,7-DHT in the cat retina were a few ganglion cells and displaced amacrine cells located in the ganglion cell layer (GCL), and some amacrine cells in the inner nuclear layer (INL). The cell density (per unit area, 1 mm2) of the 5,7-DHT accumulating cells in the GCL and INL was 910 and 134 cells, respectively. Most 5,7-DHT accumulating cells showed dark degeneration characterized by widening of the cellular organelles at early stage, and by darkening of the cytoplasm at a late stage. In addition, amacrine cells, showing a typical filamentous degeneration, were observed in a few cases. The degenerated neurons were phagocytosed by microglial cells and astrocytes. The immunoreactivity for glial fibrillary acidic protein (GFAP) in Muller cells was increased at early stage, but thereafter abruptly decreased. In a few cases, severe degenerative changes were observed in Miller cells. These results indicate that 5,7-DHT induces severe dark degeneration of IACs, and most degenerated cells could be eliminated by microglial cells and astrocytes in the cat retina. PMID- 9741548 TI - Sinus histiocytosis with massive lymphadenopathy. Multiple skull involvements. AB - Sinus histiocytosis with massive lymphadenopathy is a non-neoplastic self limiting disease of the bone marrow stem cell origin. It is characterized by painless, bilateral cervical lymphadenopathy accompanied by fever, leukocytosis, elevated erythrocyte sedimentation rate and hypergammaglobulinemia. Extranodal involvement including bone is rare. The patient is a 45-year -old female with multiple punch out lesions on her skull. MRI findings included iso-signal intensity mass at the diploid space on T1 weighted image and on T2 weighted image, mild high signal intensity was obtained. Histologically, the lesion showed proliferation of histiocytes in the fibroblastic background with formation of reactive germinal centers and many plasma cells. The histiocytes show round nuclei and occasional nucleoli and abundant cytoplasms. In area, there is lymphocytophagocytosis. Immunohistochemically, the histiocytes were positive for S-100 protein and lysozyme. PMID- 9741549 TI - Well-differentiated osteosarcoma of the rib. AB - Well-differentiated osteosarcoma is extremely rare, there is no sex predominance and the mean age is in the third decade. The tumor has a strong predilection for the long bones of the extremities, especially the metaphysis but may also occur in the diaphysis. Radiologically, the lesion shows no distinctive features, often simulating fibrous dyplasia or desmoplastic fibroma. We report a case of well differentiated osteosarcoma involving the rib of a 45-year-old female. This is a peculiar case of well differentiated osteosarcoma involving an unusual site and older patient. We also discuss histological differential diagnosis as well as clinical features of this rare disease. PMID- 9741550 TI - Fracture originated at the tip of heterotopic ossification of femur by minor external force: report of two cases. AB - Fracture of femur without evidence of severe osteoporosis rarely occurs as a result of minor external force such as a gentle passive or an active range of motion exercises. We report two cases of femoral shaft fracture occurred at the tip of heterotopic ossification of femur by a minor external force, in which the involved femur shows no evidence of remarkable osteoporosis. The possible mechanism inferred by authors is as follows: 1) heterotopic ossification limits range of motion of the hip joint; 2) a new lever arm is formed at the tip of heterotopic ossification where energy can be concentrated; 3) therefore, fractures develop even by a minor external force. Search for similar cases and further discussions on possible mechanisms and prevention of femoral fractures in patients with heterotopic ossification will be necessary. PMID- 9741551 TI - Granulocytic sarcoma as isolated extramedullary relapse after donor lymphocyte infusion in a patient with CML who relapsed after allogeneic bone marrow transplantation: a case report. AB - Isolated granulocytic sarcoma (GS) has rarely been reported in a patient who underwent allogeneic bone marrow transplantation (BMT) for chronic myelogenous leukemia (CML). We report here a patient who developed an isolated GS after achieving hematologic and cytogenetic remission by donor lymphocyte infusion for the relapse of CML following BMT. The size of GS was slightly decreased after local irradiation of 1,500 cGy without further systemic chemotherapy or immunotherapy. He remained in hematologic and cytogenetic remission without systemic relapse of CML for 8 months. Thereafter, he died of sepsis. The appropriate treatment of GS and impact of its occurrence on prognosis following allogeneic BMT has yet to be determined. PMID- 9741553 TI - Desmoplastic cerebral astrocytoma of infancy: a case report. AB - We present a case of desmoplastic cerebral astrocytoma of infancy (DCAI) in a 9 month-old boy including immunohistochemical and proliferative activity studies. It was mainly composed of glial fibrillary acidic protein (GFAP)-positive astrocytes and desmoplastic stroma. Studies with Ki-67 and synthetic phase fraction disclosed a low proliferative activity. Flow cytometric study revealed diploidy pattern. These findings suggest a positive correlation with the favorable prognosis. PMID- 9741552 TI - Manganese induced parkinsonism: a case report. AB - Manganese (Mn) intoxication is known to induce parkinsonism. Mn-induced parkinsonism preferentially affect the globus pallidus in contrast to idiopathic parkinsonism where degeneration predominantly involves the nigral pars compacta. We describe a 51-year-old man who had been occupationally exposed to Mn. He had parkinsonian features including masked face, resting tremor, and bradykinesia. He also had a cock walk and a particular propensity to fall in a backward gait. There was no sustained therapeutic response to levodopa. A fluorodopa PET scan was normal. This case indicates that Mn-induced parkinsonism can be differentiated from idiopathic parkinsonism in that the former has unique clinical features and a normal fluorodopa PET scan. PMID- 9741554 TI - Temporal lobe epilepsy caused by intrahippocampal calcified cysticercus: a case report. AB - The major differential diagnoses of epilepsy associated with small solitary lesion are tuberculous or cysticercus granuloma which are enhanced in CT and/or MRI study. We report of a 47-year-old man with intractable temporal lobe epilepsy as the presenting feature of a solitary intrahippocampal calcified mass without enhancement, which turned out to be a Taenia solium cysticercus. There was no apparent evidence of systemic cysticercosis. Imaging studies revealed a small solitary intrahippocampal calcification without perilesional enhancement, and atrophy of the hippocampal head portion. Cysticercosis should be considered as an etiology in the differential diagnosis in lesional medial temporal lobe epilepsy even without perilesional enhancement. PMID- 9741555 TI - Acute transverse myelitis caused by Coxsackie virus B4 infection: a case report. AB - Acute transverse myelitis is a rare clinical manifestation of Coxsackie virus infection which cause acute and progressive debilitating illness associated with loss of spinal cord function in the affected patients. A 62 year-old female developed symptoms of rapidly progressive paraplegia with sensory loss. On spinal MRI, T2 sagittal image showed increased signal intensity with cord swelling at T11-L2 level and 8 folds or greater rise of Coxsackie virus B4 neutralizing antibody titers was observed in the CSF. There is only one previous report of acute transverse myelitis caused by Coxsackie virus B4 infection to our knowledge. The presence of specific viral antibody titers change in the CSF and a corresponding spinal cord lesion are sufficient to suggest a causal relationship between the virus and the illness. This article is a case report of an unusual acute transverse myelitis caused by Coxsackie virus B4 infection. PMID- 9741556 TI - Religious coping and health status in medically ill hospitalized older adults. AB - Associations between specific religious coping (RC) behaviors and health status in medically ill hospitalized older patients were examined and compared with associations between nonreligious coping (NRC) behaviors and health status. The sample consisted of 577 patients age 55 or over consecutively admitted to the general medical inpatient services of Duke University Medical Center (78%) or the Durham VA Medical Center (22%). Information was gathered on 21 types of RC, 11 types of NRC, and 3 global indicators of religious activity (GIRA). Health measures included multiple domains of physical health, depressive symptoms, quality of life, stress-related growth, cooperativeness, and spiritual growth. Demographic factors, education, and admitting hospital were control variables. "Negative" and "positive" types of religious coping were identified. Negative RC behaviors related to poorer physical health, worse quality of life, and greater depression were reappraisals of God as punishing, reappraisals involving demonic forces, pleading for direct intercession, and expression of spiritual discontent. Coping that was self-directed (excluding God's help) or involved expressions reflecting negative attitudes toward God, clergy, or church members were also related to greater depression and poorer quality of life. Positive RC behaviors related to better mental health were reappraisal of God as benevolent, collaboration with God, seeking a connection with God, seeking support from clergy/church members, and giving religious help to others. Of 21 RC behaviors, 16 were positively related to stress-related growth, 15 were related to greater cooperativeness, and 16 were related to greater spiritual growth. These relationships were both more frequent and stronger than those found for NRC behaviors. Certain types of RC are more strongly related to better health status than other RC types. Associations between RC behaviors and mental health status are at least as strong, if not stronger, than those observed with NRC behaviors. PMID- 9741557 TI - Physical symptom trajectories following trauma exposure: longitudinal findings from the normative aging study. AB - This study modeled physical symptom trajectories from ages 30 to 75 in 1079 older male military veterans who were assessed every 3 to 5 years since the 1960s. Combat exposure and noncombat trauma were used to define four groups: no trauma (N = 249), noncombat trauma only (N = 333), combat only (N = 152), and both combat and noncombat trauma (N = 345). Number of symptoms on the Cornell Medical Index physical symptom scale increased 29% per decade. Men who had experienced either combat or noncombat trauma did not differ from nonexposed men, but those who had experienced both combat and noncombat trauma had 16% more symptoms across all ages. There were no differences in age-related trajectories as a function of trauma history. In cross-sectional analysis, men with combat and noncombat trauma had more posttraumatic stress disorder symptoms, but not more depression symptoms, than men with either no trauma or noncombat trauma only. Discussion focuses on the importance of considering physical as well as psychological outcomes of exposure to traumatic events. PMID- 9741558 TI - Convergent and discriminant validity of overall defensive functioning. AB - We examined the validity of the construct of overall defensive functioning and its discrimination from standard diagnostic assessments. Within a multisite field trial, patients received intake diagnostic interviews by clinicians who made standard axis I through V diagnoses, then rated defense mechanisms using the Defense Mechanism Rating Scales (DMRS). Patients filled out self-report measures of distress and defenses, the SCL-90-R, and Defense Style Questionnaire (DSQ). Overall defensive functioning (ODF) scales were derived from both the DMRS and the DSQ. Overlap between clinical and self-report ratings of defenses was modest. By two different methods of factor analysis, followed by confirmatory factor analysis, clinical ratings of ODF were clearly discriminable from axis I, axis II personality disorders, current and usual global functioning, and subjective distress. ODF measured by the DSQ was not clearly discriminated from subjective distress ratings, consistent with the hypothesis that subjective distress may distort conscious derivatives of actual defensive processes. The DSQ alone probably should not be considered as a substitute for observer-rated assessment of defensive functioning, although further study of the issue is warranted. PMID- 9741559 TI - The detection and measurement of depersonalization disorder. AB - Depersonalization disorder comprises one of the four major dissociative disorders and yet remains poorly studied. There are no reports describing the application of dissociation scales to this population. Our goal was to investigate the applicability of four such scales to depersonalization disorder and to establish screening criteria for the disorder. Two general dissociation scales and two depersonalization scales were administered to 50 subjects with DSM-III-R depersonalization disorder and 20 healthy control subjects. The depersonalization disorder group scored significantly higher than the normal control group in all scales and subscales. Factor analysis of the Dissociative Experiences Scale (DES) yielded three factors as proposed previously, absorption, amnesia, and depersonalization/derealization. A DES cutoff score of 12, markedly lower than those previously proposed for the screening of other dissociative disorders, is required for the sensitive detection of depersonalization disorder. Alternatively, the DES pathological dissociation taxon (DES-taxon) score recently generated in the literature appears more sensitive to the detection of depersonalization disorder and is better recommended for screening purposes. The other three scales were fairly strongly correlated to the DES, suggesting that they may measure similar but not identical concepts, and cutoff scores are proposed for these scales also. General implications for the screening and quantification of depersonalization pathology are discussed. PMID- 9741560 TI - The dose-effect relationships between torture and psychiatric symptoms in Vietnamese ex-political detainees and a comparison group. AB - The purpose of this study was to determine in Vietnamese ex-political detainees newly arrived into the United States a) the prevalence of torture and psychiatric symptoms and b) the dose-effect relationships between cumulative torture experience and the psychiatric symptoms of posttraumatic stress disorder (PTSD) and major depression. The study population included Vietnamese ex-political detainees (N = 51) and a comparison group (N = 22). All respondents received culturally validated instruments with known psychometric properties including Vietnamese versions of the Hopkins Symptom Checklist-25 and the Harvard Trauma Questionnaire. The ex-political detainees, in contrast to the comparison group, had experienced more torture events (12.2 SD = 4.2 vs. 2.6 SD = 3.1) and had higher rates of PTSD (90% vs. 79%) and depression (49% vs. 15%). Dose-effect relationships between cumulative torture experience and psychiatric symptoms were positive with the PTSD subcategory of "increased arousal" revealing the strongest association. These findings provide evidence that torture is associated with psychiatric morbidity in Vietnamese refugees. The demonstration of significant dose-effect responses supports the hypothesis that torture is a major risk factor in the etiology of major depression and PTSD. The generalizability of these results to other torture survivor groups is unknown. The interaction between torture and other pre- and post-migration risk factors over time in different cultural settings still needs to be examined. PMID- 9741561 TI - Three forms of somatization presenting in primary care settings in Spain. AB - The objective of this paper is to study the prevalence and clinical characteristics of functional, hypochondriacal, and presenting somatization (FSTS, HSTS, and PSTS, respectively) defined by standardized criteria, as well as the validity of their distinction in primary care in Spain. A two-stage epidemiological study of a representative sample (N = 1559) of primary care patients was carried out. In the first phase, the validated Spanish versions of General Health Questionnaire, Mini-Mental State Examination, and CAGE were used. In the second phase, the Standardized Polyvalent Psychiatric Interview, an interview for the multiaxial assessment of medical patients, was employed. The prevalence of any form of somatization in Spain was 21.3% (FSTS: 16.2%, PSTS: 9.4%, HSTS: 6.7%). Overlap of any of the three clinical forms was very frequent (42.7%). FSTS patients tended to be more chronic and showed higher scores in fatigue but lower scores in both depression and anxiety. Chronicity was frequent among somatizers, particularly in those who fulfilled more than one kind of somatization. Differences in diagnostic distribution among the three groups were also observed. In conclusion, this is the first study giving support to the validity of the distinction among three types of somatization in Spain, but overlap was more frequent than reported in North American studies. PMID- 9741562 TI - Right hemisphere, white-matter learning disabilities associated with depression in an adolescent and young adult psychiatric population. AB - Four hundred and eighty-four adolescents and young adults at an inpatient psychiatric facility were diagnosed as nonverbal learning disabled, verbal learning disabled, general learning disabled, or normal psychiatric controls. The nonverbal learning disabled group had the highest incidence of depression and was clearly different from the reading disabled group (66.3% vs. 33.3%). Nondisabled subjects were significantly more likely than the other subjects to be diagnosed with adjustment problems. Depressed subjects were significantly younger and more likely to be female. This study supports the contention that right-hemisphere, white-matter, arithmetic-disabled adolescents and young adults in a psychiatric population are at greater risk for depression than are psychiatric patients not showing this pattern. PMID- 9741563 TI - The influence of marital quality and attachment styles on traumatic grief and depressive symptoms. AB - Few studies have examined the relationship between marital quality and adjustment to the impending loss of a terminally ill spouse. Most studies of marital quality and grief have been based on retrospective reports of the marriage rather than pre-loss assessments. Here, we tested the pre-loss cross-sectional effects of having a security-enhancing marriage on traumatic grief and depressive symptoms among 59 caregivers aged 50 and over of terminally ill spouses. We also examined whether insecure attachment styles were associated with traumatic grief and depressive symptoms. Findings suggest that security-increasing marriages and insecure attachment styles put spouses at risk for elevated traumatic grief symptoms. Results also indicate that marital quality and attachment style did not interact and that neither was significantly associated with depressive symptoms. The differences in the relationship of marital quality and attachment styles to the two outcomes suggest that the etiology of traumatic grief and depressive symptoms may be distinct. PMID- 9741564 TI - Posttraumatic stress symptoms among U.S. navy divers recovering TWA flight 800. PMID- 9741565 TI - Suicidal ideation in panic disorder patients. PMID- 9741566 TI - Inpatient versus day treatment for substance abusing adolescents. PMID- 9741567 TI - Primary pulmonary hypertension: insights into pathogenesis from epidemiology. AB - Primary pulmonary hypertension (PPH) is a rare disease that affects young people predominantly of female gender. Early epidemiologic studies have shown that the diagnosis is usually made 1 to 2 years after symptoms onset, and the mean survival is reduced to 2 to 3 years thereafter. New insights into the pathogenesis of PPH by epidemiologic studies may be obtained through the utilization of informatic technologies coupled to a clear definition of the disease. Early stages of precapillary pulmonary hypertension could be identified through screening tests like echocardiography in populations with higher incidence, such as familial PPH and the conditions associated with pulmonary hypertension. These latter conditions are hemodynamically and pathologically similar to the primary form, and they can give insight into several possible aspects of the pathogenesis of PPH. Prospective registries are very useful in coordinating the collection of epidemiologic data, and new technologies, such as informatics, may improve the management and the continuous updating of the databases. PMID- 9741569 TI - A role for potassium channels in smooth muscle cells and platelets in the etiology of primary pulmonary hypertension. AB - Plasma serotonin levels are markedly elevated in patients with primary pulmonary hypertension (PPH) and platelet levels of serotonin are low. Furthermore, plasma serotonin levels remain elevated after bilateral lung transplantation, in the absence of any pulmonary hypertension. Dexfenfluramine can cause the anorexigen induced form of PPH that is clinically and histologically indistinguishable from PPH. We find that dexfenfluramine releases serotonin from platelets and inhibits its reuptake. These observations suggest that serotonin might be involved in, or be a marker for, the mechanism responsible for both forms of PPH. Dexfenfluramine causes inhibition of voltage-sensitive potassium (Kv) channels, membrane depolarization, and calcium entry in pulmonary artery smooth muscle cells and vasoconstriction in isolated perfused rat lungs. We have recently found that dexfenfluramine also inhibits Kv channels in megakaryocytes, the stem cell for platelets. In smooth muscle cells, taken from the pulmonary arteries of PPH patients, Kv channels appear to be dysfunctional. The underlying defect in PPH is likely to be an abnormality of one or more Kv channels in both pulmonary artery smooth muscle cells and platelets. Relatively few patients exposed to dexfenfluramine develop PPH. The factors responsible for susceptibility might be a difference in expression of potassium channels and/or a decrease in the endogenous production of nitric oxide. PMID- 9741568 TI - Primary pulmonary hypertension associated with the use of fenfluramine derivatives. AB - Fenfluramine derivatives (Fds) are a well-established risk factor for primary pulmonary hypertension (PPH). We compared 62 Fd-PPH patients (61 women) evaluated in our center between 1986 and 1997 with 125 sex-matched PPH patients nonexposed to Fd referred during the same period (control PPH). In the Fd-PPH group, 33 patients (53%) used dexfenfluramine alone, 7 patients (11%) used fenfluramine alone, and 5 patients (8%) used both drugs. In 17 cases (27%), Fd use was associated with that of amphetamines. Most of the exposed patients used Fd for at least 3 months (81%). The interval between the onset of dyspnea and that of drug intake was 49+/-68 months (27 days to 23 years). At the time of diagnosis, Fd-PPH and control PPH were similar in terms of New York Heart Association functional class and symptoms. The two groups significantly differed only in terms of age (50+/-12 vs 40+/-14 years) and body mass index (28+/-6 vs 23+/-4). The two groups displayed similar severe baseline hemodynamics (total pulmonary vascular resistance: 32+/-12 vs 31+/-12 IU/m2), but the percentage of responders to acute vasodilator testing was higher in control PPH (27% vs 10%, p < 0.01). As a result, more patients were treated with oral vasodilators in the control PPH group (36% vs 16%, p < 0.01) and long-term epoprostenol infusion was more frequently used in the Fd-PPH group (52% vs 31%, p < 0.01). Overall survival was similar in the two groups with a 3-year survival rate of 50%. PMID- 9741570 TI - Lipid mediator dysregulation in primary pulmonary hypertension. AB - The characteristic arteriopathy of primary pulmonary hypertension (PPH) with attendant endothelial dysfunction provides an opportunity for enhanced cellular activation in the lung. Data from many laboratories support the concept of altered eicosanoid metabolism in PPH. Rigorously quantitative measurements of the excretion of metabolites of thromboxane A2 and prostacyclin support persistent platelet activation and inadequate endothelial response in patients with PPH. Recent studies measuring excretion of prostaglandin D2 metabolites suggest that additional cell sources, such as activated tissue macrophages, may also play a role in the observed elevation in thromboxane excretion and possibly in the pathogenesis of the vascular remodeling. Additional research examining in vivo cell activation in patients receiving therapy with long-term infusion of prostacyclin may further our understanding of the pathogenesis of PPH. PMID- 9741571 TI - Nitric oxide and endothelin-1 in pulmonary hypertension. AB - BACKGROUND: We have shown previously increased expression of the potent vasoconstrictor peptide endothelin-1 (ET-1) in the pulmonary arteries of patients with pulmonary hypertension. We also demonstrated diminished expression of endothelial nitric oxide synthase, the enzyme responsible for generating nitric oxide (NO), in patients with the same disease. STUDY OBJECTIVE: To determine the expression of neuronal nitric oxide synthase (NOS-I) and endothelin-converting enzyme-1 (ECE-1) in lungs of patients with pulmonary hypertension. METHODS: Immunohistochemistry with avidin-biotin-peroxidase method. RESULTS: There was little immunostaining for NOS-I in the pulmonary arteries of normal control or diseased lungs. Moderate diffuse staining was seen in the airway epithelium and nerve bundles. Immunoreactivity for ECE-1 was seen in the airway epithelium, smooth muscle cells, and scattered macrophages of both normal and diseased lungs. Strong immunoreactivity for ECE-1 was seen in the endothelium of diseased pulmonary arteries of patients with pulmonary hypertension. CONCLUSION: We conclude that expression of NOS-I appears to be similar in normal and diseased lungs, while abundant expression of ECE-1 is present in diseased vessels, which may contribute to the pathogenesis of arteriopathy in pulmonary hypertension. PMID- 9741572 TI - Elastase and the pathobiology of unexplained pulmonary hypertension. AB - Our laboratory has focused on the increased activity of an endogenous vascular elastase in the pathobiology of pulmonary hypertension and on the mechanisms by which it is upregulated and by which it orchestrates abnormal remodeling of the vessel wall, specifically the induction of growth factors, the induction of the glycoprotein tenascin, which amplifies the proliferative response, and fibronectin, which is critical to the process of smooth muscle migration in the context of neointimal formation. We explore strategies by which targetting these processes might arrest progression or induce regression of pulmonary vascular disease associated with unexplained pulmonary hypertension. PMID- 9741573 TI - Primary pulmonary hypertension between inflammation and cancer. AB - We believe that the monoclonal cell expansion in primary pulmonary hypertension is the result of autonomous growth of stem cell-like endothelial cells, whereas the polyclonal proliferation in secondary pulmonary hypertension occurs as a response of endothelial cells to exogenous stimuli (like viral infection or high shear stress). In this context, we propose that different transcriptional and translational events govern the growth and expansion of monoclonal when compared with polyclonal pulmonary endothelial cells. The availability of antibodies directed against specific tyrosine kinase proteins involved in vasculogenesis/angiogenesis now permits the identification and localization of the components of such a misguided angiogenesis cell proliferation program in the pulmonary hypertensive vascular lesions. PMID- 9741574 TI - Genetics and immunogenetic aspects of primary pulmonary hypertension. AB - Primary pulmonary hypertension (PPH), also referred to as unexplained or idiopathic pulmonary hypertension, is the clinical term used to describe a condition in patients for which we can find no underlying cause. Patients with PPH not uncommonly also have evidence of immune dysregulation: autoimmune disorders, drug therapy, or HIV infections. We will review these associations and possible relevant abnormalities in immune regulation with regard to how they may play a role in the pathogenesis of PPH. Autoantibody-HLA correlations have been observed in several subsets of PPH patients. In addition, a familial form of PPH has been described and characterized with linkage to chromosome 2q31-q32. The identification of a specific gene for PPH and the subsequent understanding of its effects will help us identify the basic cause of PPH. Furthering our understanding regarding the role(s) and significance of immunogenetic as well as genetic aspects of the pathogenesis and pathophysiology of PPH should also lead to improved therapeutic modalities for PPH. PMID- 9741575 TI - Clinical insights into the pathogenesis of primary pulmonary hypertension. AB - Because of the lack of adequate animal models, much of our knowledge of the pathogenesis of primary pulmonary hypertension has come from clinical experiences. The clinical response to vasodilators, prostenoids, and anticoagulants as treatments appear to correlate with the pathologic changes of medial hypertrophy, intimal proliferation, and thrombosis. Endothelial dysfunction, as a primary abnormality in primary pulmonary hypertension, provides an explanation for the pathologic and clinical expression of the disease in its various forms. Other clinical features of the disease, such as age of onset and rapidity of progression, may be influenced by triggers of the disease process and underlying individual genetic susceptibility. As we have been able to correlate the spectrum of clinical observations with advances in vascular biology, newer, more focused and effective therapies should begin to emerge. PMID- 9741576 TI - Etiology and pathogenesis of primary pulmonary hypertension: a perspective. AB - In recent years, considerable advances have been made in treating primary pulmonary hypertension (PPH). These have provided a series of therapeutic options, ranging from the oral administration of calcium channel blockers to the continuous infusion of prostacyclin and/or lung transplantation. These therapeutic advances have highlighted the need for the better understanding of etiology and pathogenesis. Among the key uncertainties, the following are defined as leading uncertainties: (1) the nature of the initiating lesion; (2) the shared pathogenetic mechanisms that culminate in the pathologic lesions of PPH; (3) the molecular genetic bases for familial PPH and for susceptibility to PPH; (4) understanding of the obliterative-proliferative occlusive process in the small muscular pulmonary arteries; and (5) redefinition of "primary" and "secondary," ie, a revised nomenclature of pulmonary hypertension. A revised classification based on etiology is presented. PMID- 9741577 TI - The basic chemistry of nitrogen monoxide and peroxynitrite. AB - After a discussion of the physical chemistry of nitrogen monoxide, such as solubility (1.55 mM at 37 degrees C and an ionic strength of 0.15 M) and diffusion constant (4.8 x 10(-5) cm2s(-1)), several reactions that can acts as sinks are discussed, namely the reaction with dioxygen, with thiols and with superoxide. Of these, the latter reaction leads to a powerful oxidant, peroxynitrite. The thermodynamic and kinetic properties of this molecule are also reviewed. PMID- 9741578 TI - Oxidative chemistry of nitric oxide: the roles of superoxide, peroxynitrite, and carbon dioxide. AB - The roles of superoxide (O2.-), peroxynitrite, and carbon dioxide in the oxidative chemistry of nitric oxide (.NO) are reviewed. The formation of peroxynitrite from .NO and O2.- is controlled by superoxide dismutase (SOD), which can lower the concentration of superoxide ions. The concentration of CO2 in vivo is high (ca. 1 mM), and the rate constant for reaction of CO2 with -OONO is large (pH-independent k = 5.8 x 10(4) M(-l)s(-1)). Consequently, the rate of reaction of peroxynitrite with CO2 is so fast that most commonly used scavengers would need to be present at very high, near toxic levels in order to compete with peroxynitrite for CO2. Therefore, in the presence of physiological levels of bicarbonate, only a limited number of biotargets react directly with peroxynitrite. These include heme-containing proteins such as hemoglobin, peroxidases such as myeloperoxidase, seleno-proteins such as glutathione peroxidase, proteins containing zinc-thiolate centers such as the DNA-binding transcription factors, and the synthetic antioxidant ebselen. The mechanism of the reaction of CO2 with OONO produces metastable nitrating, nitrosating, and oxidizing species as intermediates. An analysis of the lifetimes of the possible intermediates and of the catalysis of peroxynitrite decompositions suggests that the reactive intermediates responsible for reactions with a variety of substrates may be the free radicals .NO2 and CO3.-. Biologically important reactions of these free radicals are, for example, the nitration of tyrosine residues. These nitrations can be pathological, but they also may play a signal transduction role, because nitration of tyrosine can modulate phosphorylation and thus control enzymatic activity. In principle, it might be possible to block the biological effects of peroxynitrite by scavenging the free radicals .NO2 and CO3.-. Because it is difficult to directly scavenge peroxynitrite because of its fast reaction with CO2, scavenging of intermediates from the peroxynitrite/CO2 reaction would provide an additional way of preventing peroxynitrite-mediated cellular effects. The biological effects of peroxynitrite also can be prevented by limiting the formation of peroxynitrite from .NO by lowering the concentration of O2.- using SOD or SOD mimics. Increased formation of peroxynitrite has been linked to Alzheimer's disease, rheumatoid arthritis, atherosclerosis, lung injury, amyotrophic lateral sclerosis, and other diseases. PMID- 9741579 TI - Modulation of leukocyte-endothelial interactions by reactive metabolites of oxygen and nitrogen: relevance to ischemic heart disease. AB - Ischemia and reperfusion (I/R) are thought to play an important role in the pathophysiology of ischemic diseases of the heart. It is now well appreciated that leukocyte-endothelial cell interactions are important determinants for I/R induced microvascular injury and dysfunction. There is a growing body of experimental data to suggest that reactive metabolites of oxygen and nitrogen are important physiological modulators of leukocyte-endothelial cell interactions. A number of investigators have demonstrated that I/R enhances oxidant production within the microcirculation resulting in increases in leukocyte adhesion and transendothelial cell migration. Several other studies have shown that exogenous nitric oxide (NO) donors may attenuate leukocyte and platelet adhesion and/or aggregation in a number of different inflammatory conditions including I/R. The objective of this review is to discuss the physiological chemistry of reactive metabolites of oxygen and nitrogen with special attention given to those interactions that may modulate leukocyte-endothelial cell interactions, provide an overview of the evidence implicating reactive metabolites of oxygen and nitrogen as modulators of leukocyte-endothelial cell interactions in vivo, and discuss how these mechanisms may be involved in the pathophysiology of ischemic heart disease. PMID- 9741580 TI - Chemical biology of nitric oxide: Insights into regulatory, cytotoxic, and cytoprotective mechanisms of nitric oxide. AB - There has been confusion as to what role(s) nitric oxide (NO) has in different physiological and pathophysiological mechanisms. Some studies imply that NO has cytotoxic properties and is the genesis of numerous diseases and degenerative states, whereas other reports suggest that NO prevents injurious conditions from developing and promotes events which return tissue to homeostasis. The primary determinant(s) of how NO affects biological systems centers on its chemistry. The chemistry of NO in biological systems is extensive and complex. To simplify this discussion, we have formulated the "chemical biology of NO" to describe the pertinent chemical reactions under specific biological conditions. The chemical biology of NO is divided into two major categories, direct and indirect. Direct effects are defined as those reactions fast enough to occur between NO and specific biological molecules. Indirect effects do not involve NO, but rather are mediated by reactive nitrogen oxide species (RNOS) formed from the reaction of NO either with oxygen or superoxide. RNOS formed from NO can mediate either nitrosative or oxidative stress. This report discusses various aspects of the chemical biology of NO relating to biological molecules such as guanylate cyclase, cytochrome P450, nitric oxide synthase, catalase, and DNA and explores the potential roles of NO in different biological events. Also, the implications of different chemical reactions of NO with cellular processes such as mitochondrial respiration, metal homeostasis, and lipid metabolism are discussed. Finally, a discussion of the chemical biology of NO in different cytotoxic mechanisms is presented. PMID- 9741581 TI - Retinal abnormalities in experimental vitamin E deficiency. AB - Physiological and biochemical studies have been carried out longitudinally over a period of 12 months in vitamin E deficient and control rats to gain an understanding of the mechanism whereby vitamin E conserves normal retinal function. Electroretinographic studies indicated that the primary effect of vitamin E deficiency was on the photoreceptors. Ultrastructural studies, however, did not show any morphological changes to the photoreceptors which could explain receptor dysfunction. A 30-40% loss of vitamin A (retinol) was found to be associated with vitamin E deficiency. This could be corrected by repletion with vitamin E, but there was no associated improvement in visual function. An irreversible loss of the long-chain polyunsaturated fatty acids from the retina, increased lipid peroxidation and alterations in membrane fluidity were also detected during vitamin E deficiency. We suggest that a deficiency of vitamin E leads to changes in the membrane microenvironment, which could affect photo transduction by either impairing the ability of rhodopsin to undergo conformational changes to the active form, or by disrupting the hyperpolarising and depolarising processes of the photoreceptors. PMID- 9741582 TI - Peroxynitrite modulates MnSOD gene expression in lung epithelial cells. AB - Peroxynitrite (ONOO-) is a strong oxidant derived from nitric oxide ('NO) and superoxide (O2.-), reactive nitrogen (RNS) and oxygen species (ROS) present in inflamed tissue. Other oxidant stresses, e.g., TNF-alpha and hyperoxia, induce mitochondrial, manganese-containing superoxide dismutase (MnSOD) gene expression. These experiments tested whether ONOO regulated MnSOD gene expression in human lung epithelial (A549) cells. 3-morpholinosydnonimine HCI (SIN-1) (10 or 1000 microM) increased MnSOD mRNA, but did not change hypoxanthine guanine phosphoribosyl transferase (HPRT) mRNA. Authentic peroxynitrite (ONOO ) (100-500 microM) also increased MnSOD mRNA but did not change constitutive HPRT mRNA expression. ONOO stimulated luciferase gene expression driven by a 2.5 kb fragment of the rat MnSOD gene 5' promoter region. MnSOD gene induction due to ONOO- was inhibited effectively by L-cysteine (10 mM) and partially inhibited by N-acetyl cysteine (50 mM) or pyrrole dithiocarbamate (10 mM). .NO from 1 propanamine, 3-(2-hydroxy-2-nitroso-1-propylhydrazine) (PAPA NONOate) (100 or 1000 microM) did not change MnSOD or HPRT mRNA. Neither H202 nor NO2-, breakdown products of SIN-1 and ONOO , had any effect on MnSOD mRNA expression; however, ONOO- and SIN-1 did not increase MnSOD protein content detectable by western blots, nor did they increase MnSOD enzymatic activity. Increased steady state [O2.-] in the presence of .NO yields ONOO , and ONOO has direct, stimulatory effects on MnSOD transcript expression. PMID- 9741583 TI - A practical assay of lipoate in biologic fluids and liver in health and disease. AB - A procedure for assaying lipoic acid concentration in biologic fluids and tissues was devised using a eukaryotic protozoan Tetrahymena thermophila. T.thermophila has a specific and sensitive (30 pg/ml) requirement for lipoic acid. Unlike humans and other microorganisms, T.thermophila can not synthesize lipoic acid; hence, its requirement for exogenous lipoic acid is specific. The lipoic acid supplied to T. thermophila by the processing of biologic fluids and tissues during the assay procedure, permits the derivation of a practical assay for lipoate concentration as described here. Lipoate concentration in biologic fluids and tissue obtained from healthy humans, compared to those obtained from patients with renal and liver disease, indicate deviations from normal during disease. Absorption chartings of 200 mg of DL-alpha-lipoic acid in humans indicate a peak concentration of lipoate in plasma 2 h after ingestion and then a steady descent of lipoate to a baseline level after 24 h. With this practical assay, it is now possible to chart lipoate's antioxidant activity and therapeutic action during health and disease. PMID- 9741584 TI - The effect of idebenone, a coenzyme Q analogue, on hydrophobic bile acid toxicity to isolated rat hepatocytes and hepatic mitochondria. AB - Oxidant stress induced by hydrophobic bile acids has been implicated in the pathogenesis of liver injury in cholestatic liver disorders. We evaluated the effect of idebenone, a coenzyme Q analogue, on taurochenodeoxycholic acid (TCDC) induced cell injury and oxidant stress in isolated rat hepatocytes and on glycochenodeoxycholic acid (GCDC)-induced generation of hydroperoxides in fresh hepatic mitochondria. Isolated rat hepatocytes in suspension under 9% oxygen atmosphere were preincubated with 0, 50, and 100 micromol/l idebenone for 30 min and then exposed to 1000 micromol/l TCDC for 4 h. LDH release (cell injury) and thiobarbituric acid reactive substances (measure of lipid peroxidation) increased after TCDC exposure but were markedly suppressed by idebenone pretreatment. In a second set of experiments, the addition of 100 micromol/l idebenone up to 3 h after hepatocytes were exposed to 1000 micromol/l TCDC resulted in abrogation of subsequent cell injury and markedly reduced oxidant damage to hepatocytes. Chenodeoxycholic acid concentrations increased to 5.15 nmol/10(6) cells after 2 h and to 7.05 after 4 h of incubation of hepatocytes with 1000 micromol/l TCDC, and did not differ in the presence of idebenone. In freshly isolated rat hepatic mitochondria, when respiration was stimulated by succinate, 10 micromol/l idebenone abrogated the generation of hydroperoxides during a 90-minute exposure to 400 micromol/l GCDC. These data demonstrate that idebenone functions as a potent protective hepatocyte antioxidant during hydrophobic bile acid toxicity, perhaps by reducing generation of oxygen free radicals in mitochondria. PMID- 9741585 TI - Calcium vs. iron-mediated processes in hydrogen peroxide toxicity to L929 cells: effects of glucose. AB - H2O2 toxicity was studied in L929 cells in the presence and absence of glucose. The data obtained in the absence of glucose suggest a Ca2+-dependent mechanism of cell injury. No evidence was found for any involvement of iron in the process. In particular, cell injury was unaffected by the intracellular iron chelators 2,2' dipyridyl and deferoxamine or by the hydroxyl radical scavengers DMSO and DMPO. On the other hand, the intracellular Ca2+ chelator BAPTA/AM provided significant protection. The cytosolic Ca2+ level rapidly and consistently increased after H2O2 addition, prior to visible bleb formation and loss of cell viability. Additionally, GSH not only prevented cell death but also significantly decreased cytosolic calcium accumulation. In the presence of glucose, however, Ca2+ does not seem to play any role in H2O2 toxicity. Cell death is now mainly mediated by iron: the iron chelators and hydroxyl radical scavengers prevented cell injury, the increase in cytosolic Ca2+ was significantly less pronounced, and BAPTA/AM did not exert any protection under these conditions. Hence, the metabolic state of the L929 cells, as given by the availability of glucose, decisively determines the biochemical mechanism of H2O2 cell injury. PMID- 9741586 TI - Effect of selenium deficiency on cellular and extracellular glutathione peroxidases: immunochemical detection and mRNA analysis in rat kidney and serum. AB - To determine the effect of selenium (Se) deficiency on expression of glutathione peroxidase (GSH-Px) 1 and 2, we measured GSH-Px activity in rat serum, liver and kidneys, serum immunoreactive GSH-Px 2, and the mRNAs of kidney GSH-Px 1 and 2. We purified rat GSH-Px 2 and raised polyclonal antibodies. Immunoreactive GSH-Px 2 was measured by rocket immunoelectrophoresis. GSH-Px 2 was purified 1470-fold with a specific activity of 250 units/mg. Immunoblotting detected only GSH-Px 2 in rat serum, and much less GSH-Px 2 than GSH-Px 1 in kidney. Immunoblot signal of kidney GSH-Px 1 and 2 decreased progressively in Se deficient rats. Serum GSH Px activity in Se deficient rats at 1, 2, 3, and 4 weeks declined to 33, 20, 10, and 9% of the control, while the serum level of immunoreactive GSH-Px 2 was 58, 24, 15, and 10% of the control, suggesting the presence of an inactive protein at week 1. GSH-Px activity declined to 4 and 11% of the control in the liver and kidney at 4 weeks. The mRNAs of kidney GSH-Px 1 and 2 showed similar decreases, and were 24 and 23% of the control at 4 weeks. GSH-Px mRNA levels were better preserved than GSH-Px activity, suggesting that GSH-Px expression was regulated at both pre-translational and translational levels. PMID- 9741587 TI - Protection of PC12 cells glutathione peroxidase in L-DOPA induced cytotoxicity. AB - L-DOPA may cause side-effects during the treatment of Parkinson's disease. We investigated the role of glutathione peroxidase (GSHPx) in cellular defense against L-DOPA cytotoxicity. A line of PC12 cells overexpressing GSHPx with plasmid pRc/CMV-GSHPx was established and stable transfectants overexpressing GSHPx were used for this study. GSHPx activity was found to be 1.5-fold higher in GSHPx-transfectants than in mock-controlled transfectants. Transfectants over expressing GSHPx were also significantly more resistant to exposure to either L DOPA or t-butyl hydroperoxide than mock-transfected cells. Results suggested that L-DOPA may cause neuronal cell death by an oxidative pathway and GSHPx may play an important role in cellular defense against oxidative stress. PMID- 9741588 TI - Different mechanisms are progressively recruited to promote Cu(II) reduction by isolated human low-density lipoprotein undergoing oxidation. AB - The kinetics of Cu(II) reduction and its relationship to the process of low density lipoprotein (LDL) oxidation were investigated in isolated human LDL incubated with CuSO4 by using the Cu(I) chelator and indicator dye bathocuproine disulfonate (BC). The inclusion of BC in the incubation medium containing isolated LDL and different concentrations of CuSO4 revealed a biphasic kinetics of Cu(II) reduction consisting of an early phase followed by a plateau phase and a subsequent extensive reduction phase. The amount of Cu(I) formed during the early phase, as well as the rate of its generation, were strictly dependent on both the level of Cu(II) available (saturation was observed at 20 and 50 microM CuSO4) and the concentration of alpha-tocopherol within native LDL particles. Artificial enrichment of LDL with different concentrations of alpha-tocopherol led to a parallel increase of both the amount of Cu(II) reduced and the rate of reduction. The late phase of Cu(II) reduction was strictly related to the availability of copper but was largely independent from alpha-tocopherol. Neither the amount of Cu(I) generated nor the rate of generation were saturated at concentrations of copper up to 100 microM. Comparable results were obtained by adding BC at different time-points to the LDL-copper mixture, in order to measure at the same time-points both the true rate of Cu(II) reduction and the generation of TBARS during the dynamic process of LDL oxidation. The rate of Cu(II) reduction was already high during the lag-phase of the LDL oxidation profile and progressively decreased as alpha-tocopherol concentration decreased. The subsequent increase in the rate of Cu(II) reduction paralleled the formation of TBARS during the extensive LDL oxidation phase. These results suggest that different mechanisms of Cu(II) reduction, namely alpha-tocopherol-dependent and independent (likely lipid peroxide-dependent), are progressively recruited during copper-promoted LDL oxidation. PMID- 9741589 TI - Mechanisms of inactivation of hepatocyte protein kinase C isoforms following acute ethanol treatment. AB - Acute ethanol exposure of rat isolated hepatocytes leads to a significant decrease (-30%) in cytosolic enzymatic activity of classic protein kinase C (PKC) isoforms, while immunoreactive protein level measured by Western Blot remains unaffected. The inactivation of classic cytosolic isoforms appears dependent on the modification of the enzyme function, probably due to ethanol metabolism. In fact, pretreatment with 4-methylpyrazole (4MP), an inhibitor of alcohol dehydrogenase, fully prevented such damage. After ethanol treatment, a decrease of about 40% in both enzymatic activity and immunoreactive protein level of novel PKC isoforms was evident both in the soluble and particulate fractions. Even if 4MP cell pre-treatment afforded protection in this case too, the inhibitory action of ethanol on novel PKC hepatocyte isoforms involves a proteolytic mechanism as shown by Western Blot analysis. The reproduction of PKC inactivation by ethanol in hepatocyte lysate excluded a role of peroxisomal hydrogen peroxide in the pathogenesis of the damage investigated. This damage was not reduced by addition of catalase to the lysate model system. PMID- 9741590 TI - ESR and HPLC-EC analysis of ethanol oxidation to 1-hydroxyethyl radical: rapid reduction and quantification of POBN and PBN nitroxides. AB - Extensive ESR spin-trapping studies have shown that ethanol is oxidized to 1 hydroxyethyl radical (HER) by rat and deer mice liver microsomal systems. The ESR detection of POBN/HER nitroxide in bile, and formation of antibodies, which recognize HER adducts in alcoholics, suggest that HER is produced in vivo. In liver, where ethanol is primarily metabolized, only traces of PBN/HER nitroxide are documented. One limitation of the ESR spin-trapping technique, however, is that the nitroxides formed in the presence of cellular reductants can be metabolized to the corresponding ESR "silent" hydroxylamines. Ascorbate and NADPH plus liver microsomes were found to reduce rapidly both POBN/HER and PBN/HER nitroxides to their ESR "silent" hydroxylamine derivatives. An HPLC method with electrochemical detection was developed for the detection and quantification of both POBN/HER and PBN/HER nitroxides, as well as their hydroxylamines. Both the diastereomers of the POBN/HER nitroxide and hydroxylamine can be detected, as can both isomers of the PBN/HER nitroxide, and it is estimated that the sensitivity of the HPLC procedure is in the nM range when using EC detection. The hydroxylamines are stable in ethanol, while pH-dependent auto-oxidation occurs in aqueous buffers. Some of the characteristics associated with HER formation by microsomes as detected with ESR (e.g., sensitivity to SOD and catalase, increase after induction of CYP2E1) are reproduced with the HPLC method. By quantification of the POBN/HER hydroxylamine, the NADPH-dependent rates of HER formation by microsomes from pyrazole-treated rats are estimated to be about 1-1.5 nmol HER per min per mg microsomal protein. This rate is less, as compared to the two electron-dependent rate of acetaldehyde formation by these microsomes, about 10 15 nmol per min per mg protein. Thus, at first approximation, the one electron dependent rate of ethanol oxidation is about 10% the two electron-dependent rate by isolated microsomes. The HPLC procedure can readily detect the POBN/HER and PBN/HER nitroxides and their hydroxylamine derivatives in the same sample and may be of value in detecting HER spin-trapped adducts under biological reducing conditions. PMID- 9741591 TI - Liposome effect on the cytochrome c-catalyzed peroxidation of carbonyl substrates to triplet species. AB - Cytochrome c exhibits peroxidase activity on diphenylacetaldehyde (DPAA) and 3 methylacetoacetone (MAA), which is greatly affected by the presence and nature of charged liposome or micelle interfaces interacting with the enzyme. The ferricytochrome c reaction with DPAA is accelerated when the enzyme is attached to negatively charged interfaces. Whatever the medium, bulk solution or negatively charged dicetylphosphate (DCP), phosphatidylcholine/phosphatidylethanolamine/cardiolipin (PC/PE/CL) liposomes, this chemiluminescent reaction is accompanied by reduction of cytochrome c to its ferrous form. In turn, MAA is oxidized by cytochrome c exclusively when bound to DCP liposomes. Contrary to DPAA oxidation, the MAA reaction is followed by bleaching of cytochrome c, reflecting damage to the hemeprotein chromophore. The cytochrome-c-catalyzed oxidation of either DPAA or MAA leads to concomitant disappearance of the enzyme charge transfer absorption band at 695 nm. This suggests that the peroxidase activity of cytochrome c involves substrate-induced loss of the methionine ligand at the iron sixth coordination position, which is favored by interaction of cytochrome c with negatively charged interfaces. Accordingly, a decrease and blue shift of the charge transfer band could be observed in cytochrome-c-containing negatively charged DCP, PC/PE/CL liposomes or lysophosphatidylethanolamine micelles in the presence of DPAA or MAA. PMID- 9741592 TI - Oxidative damage to sarcoplasmic reticulum Ca2+-ATPase AT submicromolar iron concentrations: evidence for metal-catalyzed oxidation. AB - The sarcoplasmic reticulum (SR) calcium ATPase carries out active Ca2+ pumping at the expense of ATP hydrolysis. We have previously described the inhibition of SR ATPase by oxidative stress induced by the Fenton reaction (Fe2+ + H2O2 --> HO. + HO- + Fe3+). Inhibition was not related to peroxidation of the SR membrane nor to oxidation of ATPase thiols, and involved fragmentation of the ATPase polypeptide chain. The present study aims at further characterizing the mechanism of inhibition of the Ca2+-ATPase by oxygen reactive species at Fe2+ concentrations possibly found in pathological conditions of iron overload. ATP hydrolysis by SR vesicles was inhibited in a dose-dependent manner by micromolar concentrations of Fe2+, H2O2, and ascorbate. Measuring the rate constants of inactivation (k inact) at different Fe2+ concentrations in the presence of saturating concentrations of H2O2 and ascorbate (100 microM each) revealed a saturation profile with half maximal inactivation rate at ca. 2 microM Fe2+. Inhibition was not affected by addition of 200 microM Ca2+ to the medium, indicating that it was not related to iron binding to the high affinity Ca2+ binding sites in the ATPase. Furthermore, inhibition was not prevented by the water-soluble hydroxyl radical scavengers mannitol or dimethylsulfoxide, nor by butylated hydroxytoluene (a lipid peroxidation blocker) or dithiothreitol (DTT). However, when Cu2+ was used instead of Fe2+ in the Fenton reaction, ATPase inhibition could be prevented by DTT. We propose that functional impairment of the Ca2+-pump may be related to oxidative protein fragmentation mediated by site-specific Fe2+ binding at submicromolar or low micromolar concentrations, which may occur in pathological conditions of iron overload. PMID- 9741593 TI - Increased plasma levels of lipid hydroperoxides in patients with ischemic stroke. AB - A large body of experimental research indicates that the generation of free radicals leading to oxidative stress plays a role in the pathogenesis of ischemic brain injury, but evidence in humans is limited. We examined plasma levels of lipid hydroperoxides (measured as cholesteryl ester hydroperoxides, CEOOH) and ascorbic acid in 32 patients with cortical stroke, as compared with 13 patients with lacunar infarct. Patients with cortical stroke had significantly increased levels of CEOOH, which peaked on Day 5 after the ictus. Small decreases in ascorbic acid concentrations were not significant. There was a significant positive correlation of CEOOH with the NIH stroke scale, and a significant negative correlation with the Glasgow coma scale. Concentrations of CEOOH were significantly higher in patients with total anterior cerebral syndrome as compared with patients with partial anterior cerebral syndrome or posterior cerebral syndrome. Stroke volumes computed from CT or MRI scans were significantly correlated with plasma CEOOH levels. These findings implicate oxidative stress in ischemic brain injury in humans and suggest that measurements of CEOOH in plasma may be useful both prognostically as well as in monitoring therapeutic interventions. PMID- 9741594 TI - Cupric nitrilotriacetate induces oxidative DNA damage and apoptosis in human leukemia HL-60 cells. AB - Recent reports have implicated a possible role of reactive oxygen species (ROS) in the induction and mediation of apoptosis and DNA damage. Oxidative DNA base modification induced by cupric nitrilotriacetate (Cu-NTA) and the following apoptosis were observed in human promyelocytic leukemia HL-60 cells. We measured the level of ROS in the cells by using a fluorescence probe, 2',7' dichlorofluorescein diacetate (DCFH-DA), and the amount of a modified DNA base, 8 hydroxydeoxyguanosine (8-OHdG) by HPLC-ECD. It was found that Cu-NTA exposure significantly enhanced ROS and 8-OHdG formation in the cells. Meanwhile, we observed both DNA fragmentation and morphological changes characteristic of apoptosis, which was also determined quantitatively by flow cytometry and showed dose- and time-dependent manners. Furthermore, several antioxidants such as dimethyl sulfoxide (DMSO), superoxide dismutase (SOD), and catalase were used to detect whether the apoptosis could be blocked. Only DMSO protected against this form of cell death. To elucidate molecular events in the apoptosis, expressions of Bcl-2 protein family members, such as Bcl-2, Bcl-X and Bax, and heat shock protein 70 (HSP-70) were measured by western blotting using specific antibodies. The levels of Bax and Bcl-Xs remained largely unchanged, but the Bcl-2 and Bcl-XL expression showed down-regulation. After 24 h incubation in the presence of copper, the levels of Bcl-2 and Bcl-XL reduced about 33.8% and 51.1% compared with untreated cells, respectively. Furthermore, after 16 h incubation, the level of HSP-70 expression was about 3.4-fold greater than that in untreated cells, suggesting that HSP-70 is important in increasing resistance to oxidative stress induced by Cu-NTA. But overexpression of HSP-70 failed to protect HL-60 cells from apoptosis induced by Cu-NTA. We inferred that Cu-NTA may induce oxidative DNA damage through free radical injuries, which may turn on the apoptosis in HL 60 cells. PMID- 9741595 TI - Modulation of antioxidant responses by arsenic in maize. AB - The effects of arsenic on the expression of the antioxidant genes encoding superoxide dismutase, catalase, and glutathione S-transferase, as well as the activity of SOD and CAT enzymes, were examined at different developmental stages and in different tissues. Both CAT and SOD activities increased in response to low concentrations (0.01-0.1 mM) of arsenic in developing maize embryos. In germinating embryos both CAT and SOD activities increased in response to a wide range of arsenic concentrations (0.01-10 mM). Cat1 transcript increased in response to arsenic in developing and germinating embryos and in young leaves. Conversely, Cat2 increased at low concentrations of arsenic only in germinating embryos. Cat3 transcript levels increased in response to low concentrations of arsenic only in developing embryos. Sod3 transcript increased at low concentrations of arsenic in developing, germinating embryos and in leaves. The cytosolic Sod4 and Sod4A increased in response to arsenic in germinating embryos, while only Sod4 transcript increased in response to arsenic in leaves. Expression of Gst1 was similar to that of Cat1 in all tissues examined. These results indicate that arsenic triggers tissue and developmental stage specific defense responses of antioxidant and detoxification related genes in maize. PMID- 9741596 TI - Metal-mediated DNA damage induced by diabetogenic alloxan in the presence of NADH. AB - Alloxan is known to induce diabetes in experimental animals through destruction of insulin-producing 3-cells of pancreas. The mechanism of DNA damage induced by alloxan was investigated using 32P-labeled human DNA fragments. Cu(II)-dependent DNA damage increased with the concentration of alloxan and NADH. Alloxan induced DNA cleavage frequently at thymine and cytosine residues in the presence of NADH and Cu(II). Catalase and bathocuproine, a Cu(I)-specific chelator, almost completely inhibited DNA damage, suggesting the involvement of H2O2 and Cu(I). Alloxan induced Cu(II)-dependent production of 8-oxodG in calf thymus DNA in the presence of NADH. UV-visible and electron spin resonance (ESR) spectroscopic studies showed that superoxide anion radical and alloxan radical were generated by the reduction of alloxan by NADH, and also by the autoxidation of dialuric acid, the reduced form of alloxan. These results suggest that the copper-oxygen complex derived from the reaction of H2O2 with Cu(I) participates in Cu(II) dependent DNA damage by alloxan plus NADH and dialuric acid. The mechanism of DNA damage is discussed in relation to diabetogenic action of alloxan. PMID- 9741597 TI - Leukotoxin (9, 10-epoxy-12-octadecenoate) impairs energy and redox state of isolated perfused rat lung. AB - We investigated the perturbation of energy balance and redox state in leukotoxin (9, 10-epoxy-12octadecenoate) (Lx)- and endothelin-1 (ET-1)-induced lung injury, using isolated perfused rat lungs. To examine any relationship between these parameters, intracellular levels of adenine nucleotides, pyridine coenzymes and glutathione were determined by reversed-phase high-performance liquid chromatography (HPLC) in the freeze-dried tissues of isolated rat lungs. The tissue samples were perfused with a physiological salt solution containing either Lx only, Lx plus NG-monomethyl-L-arginine (L-NMMA), Lx plus NG-monomethyl-D arginine (D-NMMA), Lx plus superoxide dismutase (SOD) or ET-1 only. In isolated perfused lung tissue, 10 mol of Lx caused permeability-increased lung injury, and 10 nM of ET-1, which caused a comparable increase in wet lung weight, evoked pulmonary capillary hypertensive lung injury. Lx-injured lungs showed decreases in the contents of ATP, NADPH, NADH, reduced glutathione (GSH), (2ATP + ADP)/2(ATP + ADP + AMP) ratio (energy charge) and NADH/NAD+ ratio, and increased the contents of ADP and AMP compared with the vehicle control and ET-1-injured lungs. Such effects of Lx were significantly attenuated by pretreatment with 0.4 mM L-NMMA or 500 units/ml of SOD, but not with 0.4 mM D-NMMA. On the other hand, the ET-1-injured lung evidenced decreased tissue GSH. These findings indicate that Lx shifted the lung redox state toward oxidation and that Lx-induced lung injury was involved in the imbalance of the energy and redox state via production of nitric oxide and/or superoxide anion. PMID- 9741598 TI - Effect of gas-containing microspheres and echo contrast agents on free radical formation by ultrasound. AB - Stabilized microbubbles (microspheres) are widely used to enhance the contrast of ultrasound imaging. Our data provide direct evidence that the contrast agents, Levovist, PVC-AN (polyvinylidene chloride-acrylonitryl copolymer), and Albunex (compared to 5% human albumin), at concentrations comparable to those used for ultrasound imaging, enhance H2O2 production (through the superoxide-dependent pathway) in air-saturated aqueous solutions exposed to 47 kHz ultrasound above the cavitation threshold. These agents also act as scavengers of .H atoms and .OH radicals, thus lowering H2O2 formation (by recombination of .OH radicals) in argon-saturated solutions. EPR spin trapping also reveals that secondary radicals derived from the contrast agents are produced by reactions with .H and .OH which are formed by pyrolysis of water inside cavitation bubbles. In addition, the contrast agents themselves undergo pyrolysis reactions in the cavitation bubbles as demonstrated by formation of methyl radicals. Possible deleterious consequences of the formation of sonochemical intermediates may have to be assessed, particularly since some of the echo contrast agents have been shown to lower the cavitation threshold of diagnostic ultrasound. Unlike the microspheres formed from organic molecules, inorganic microspheres, Eccospheres, because of their stability and inert nature with respect to participation in free radical processes, appear to be suitable tools for enhancing the yields of aqueous sonochemical reactions. PMID- 9741599 TI - Direct observation of lipid hydroperoxides in phospholipid vesicles by electrospray mass spectrometry. AB - Positive ion electrospray ionization mass spectrometry was used to obtain a lipid profile of vesicles prepared from egg yolk lethicin and enriched with arachidonylstearoyl phosphatidylcholine and dipalmitoyl phosphatidylcholine. The vesicles were oxidized by treatment with tert-butylhydroperoxide and iron (II) sulfate, and the formation of hydroperoxides of the polyunsaturated lipid arachidonylstearoyl phosphatidylcholine was observed. The native lipid signal at 832 a.m.u. decreased and new signals appeared at 864, 896, and 928 a.m.u., corresponding to the addition of one (+32), two (+64), and three (+96) molecules of dioxygen. The dihydroperoxide was found to be the most favourable peroxide product, but it appeared that a degradation of the hydroperoxides was occurring concomitant with their formation, and only their net formation was observed. The rate of depletion of the polyunsaturated lipid and the rate of accumulation of the hydroperoxides was found to increase with the Fe2+ concentration between 10 microM and 2 mM, and was also dependent on the tert-butylhydroperoxide concentration. This is the first report of analysis of lipid hydroperoxides by electrospray mass spectrometry, showing that technique offers a sensitive, direct, and informative approach to the study of oxidative damage to biological membranes. PMID- 9741600 TI - Specific S-nitrosothiol (thionitrite) quantification as solution nitrite after vanadium(III) reduction and ozone-chemiluminescent detection. AB - Increasing evidence suggests that S-nitrosothiols (thionitrites) might represent naturally occurring nitric oxide surrogates and function as intermediates in nitrogen monoxide metabolism. A facile, sensitive, and selective micromethod has been developed and validated for quantification of S-nitrosothiols as their mercury-displaceable nitrogen monoxide content. In this method, brief (5-min), room-temperature pretreatment of S-nitrosothiol with a molar excess of aqueous mercuric chloride was used to liberate into solution, quantitatively, the nitrogen monoxide moiety, which rapidly and quantitatively converted to its stable solution end-product, nitrite. Solution nitrite was reduced back to nitric oxide with vanadium(III), and the nitric oxide was detected by gas-phase chemiluminescence after reaction with ozone in a commercial nitric oxide analyzer. A linear relationship was observed between S-nitrosothiol-bound nitrogen monoxide and ozone-chemiluminescent detector response over a wide range (16.3-3500 pmol) of nitric oxide, as generated by reaction of vanadium(III) with either nitrite standard or mercury-treated S-nitrosothiol. Assay response was quantitatively identical for equivalent amounts of nitrite and S-nitrosothiol bound nitrogen monoxide. The method displayed 96% selectivity for nitrite vs. nitrate and negligible (<2%) interference by nitrosated compounds bearing nitrogen monoxide moieties bound to either nitrogen or carbon. The lower limits of quantitative sensitivity and qualitative detection were below 50 and 20 pmol S nitrosothiol-bound nitrogen monoxide-equivalents, respectively. The intraday and interday coefficients of variation did not exceed 8%. This technique has been applied to quantify structurally diverse natural and synthetic S-nitrosothiols with quantitative recovery from complex biological samples such as culture media and plasma at levels of nitrogen monoxide-equivalents undetectable by the popular Saville colorimetric method. PMID- 9741601 TI - The case history of an elite ultra-endurance cyclist who developed chronic fatigue syndrome. AB - An elite ultra-endurance athlete, who had previously undergone physiological and performance testing, developed chronic fatigue syndrome (CFS). An incremental cycling exercise test conducted while he was suffering from CFS indicated decreases in maximum workload achieved (Wmax; -11.3%), the maximum oxygen uptake (VO2max; -12.5%), and the anaerobic threshold (AT; -14.3%) compared to pre-CFS data. A third test conducted after the athlete had shown indications of significant improvement in his clinical condition revealed further decreases in Wmax (-7.9%), VO2max (-10.2%) and AT (-8.3%). These data, along with submaximal exercise data and muscle biopsy electron microscopic analyses, suggest that the performance decrements were the result of detraining, rather than an impairment of aerobic metabolism due to CFS per se. These data may be indicative of central, possibly neurological, factors influencing fatigue perception in CFS sufferers. PMID- 9741602 TI - Phonophoresis versus ultrasound in the treatment of common musculoskeletal conditions. AB - PURPOSE: The purpose of this study was to determine whether the pain response after phonophoresis (PH) differs from the pain response after ultrasound (US) alone. METHODS: Forty-nine subjects with soft tissue injuries including epicondylitis, tendinitis, and tenosynovitis were randomly assigned (double blinded technique) to PH or US treatment groups. Both groups received 8 min of continuous US at 1.5 w x cm(-2), three times per week for 3 wk. For the PH group a gel containing 0.05% fluocinonide was used as a coupling agent. An identical gel absent the steroid was used for the US group. Subjects indicated their pain level by marking on a visual analog scale (VAS) at the start of treatment and at the end of weeks 1, 2, and 3. Pressure algometry was used to note tolerance to direct pressure over the target tissue. ANOVA for repeated measures was used to analyze data. RESULTS: At the end of 3 wk of treatment, both groups combined showed a significant decrease in pain level and an increase in pressure tolerance (P < 0.05), but there were no differences between groups from the onset of treatment to the end of week 3 (VAS: US 5.5-1.9, PH 5.0-2.0; algometry (involved limb): US 4.7 lb-7.1 lb, PH 5.1 lb-6.6 lb). CONCLUSIONS: We conclude that US results in decreased pain and increased pressure tolerance in these selected soft tissue injuries. The addition of PH with fluocinonide does not augment the benefits of US used alone. PMID- 9741603 TI - Spectral analysis of electroencephalography changes after choking in judo (juji jime). AB - PURPOSE: The present study was carried out to investigate possible electroencephalographic changes induced by choking in judo (shime-waza) by means of spectral analysis and brain mapping. METHODS: Power spectral changes in Electroencephalography (EEG) were recorded in six experienced judoka who underwent a choking trial with a "shime-waza choking" technique called juji-jime. RESULTS: A significant increase of global field power in the delta- and theta range occurred, while physiological alpha-power decreased. These changes in the low-frequency range reached a statistically significant level within a time span up to 20 s after choking, which was performed at an average choking time of 8 s. In no case did choking provoke neuropsychological symptoms. Yet, spectral EEG analysis revealed subclinical changes of brain function. CONCLUSIONS: Choking in judo may induce subclinical electroencephalographic perturbations. The extent and duration can be objectified by means of spectral analysis of EEG data, global field power computation, and brain-mapping representation. PMID- 9741604 TI - Ground reaction forces and EMG activity with ankle bracing during inversion stress. AB - PURPOSE: The purpose of this investigation was to evaluate the effects of external ankle support on ground reaction forces and myoelectrical activity of selected lower extremity muscles during dynamic inversion stress. METHODS: Twenty four healthy males performed five trials of a lateral dynamic movement at a rate between 80-90% of their maximal speed under three ankle brace conditions (no brace--control, Aircast Sport-Stirrup, Active Ankle). Ground reaction forces along the mediolateral axis and EMG activity of the peroneus longus, tibialis anterior, and medial gastrocnemius were simultaneously recorded during force plate contact. RESULTS: Ankle bracing did not affect peak impact force (P > 0.05), maximum loading force (P > 0.05), or peak propulsion force (P > 0.05) in the lateral direction compared with the control condition. Ankle bracing reduced the EMG activity of the peroneus longus during peak impact force compared with the control condition (P < 0.05), although no differences were noted between the two braces. Furthermore, peroneous longus activity during maximum loading force and peak propulsion remained unaffected (P < 0.05). Ankle bracing did not affect the EMG activity of the tibialis anterior and medial gastrocnemius at the point of peak impact force, maximum loading force (P > 0.05), and peak propulsion force (P > 0.05). CONCLUSIONS: These data suggest that ankle bracing may not affect the forces experienced at the foot and ankle, but helps reduce the strain placed on the peroneus longus during peak impact force. Furthermore, ankle bracing does not alter the function of the tibialis anterior and medial gastrocnemius during dynamic inversion stress. PMID- 9741605 TI - Comparison of injury patterns in elite hockey players using ice versus in-line skates. AB - PURPOSE: The purpose of this study was to assess the variation of injury patterns between hockey players who use in-line roller skates versus those who use ice skates. METHODS: Injury surveillance was undertaken on three professional hockey teams. Two performed on in-line skates and one performed on ice skates. Injury patterns including mechanism of injury, anatomic location, time-loss from sport, and injury type were evaluated with respect to the use of type of skate. The number of athletic exposures (AE) was calculated for each athlete to establish a relative risk. All athletes were elite professional athletes, and injuries were recorded and categorized by a certified athletic trainer or physician. RESULTS: Of the 215 games and 1122 athletic exposures evaluated, 142 injuries were recorded that required an evaluation by a physician and 46 of those required a time loss from sport. The total injury rate was similar between the two sports (in-line: 139 per 1000 AE; ice: 119 per 1000 AE) although injuries tended to be more severe in ice hockey (average time loss from sport: ice, 8.3 games; in-line, 6.5 games). CONCLUSIONS: Comparison of injury patterns by anatomic location, mechanism of injury, and injury type were similar between players using the two types of skates except that ice skates were associated with an increase in the number of lacerations, in-line skates were associated with an increased number of injuries secondary to checking and a decreased number of injuries relative to skate equipment, and ice hockey had an increased risk of head and neck injuries compared with hockey on in-line skates. PMID- 9741606 TI - Isometric and dynamic exercise studied with echo planar magnetic resonance imaging (MRI). AB - PURPOSE: The effect of different types of exercise upon echo planar (EP) magnetic resonance (MR) images was examined during and after both dynamic and isometric dorsi-flexion exercises at matched workloads and durations. METHODS: Healthy untrained subjects performed either dynamic exercise through a full range of motion and against a constant resistance or isometric exercise at the center of the range of motion and against a constant resistance at 25 or 70% their measured maximum voluntary contraction (MVC). EP MR images were acquired at 1.5 T every 4 s before (4 images), during (27 images), and after (29-65 images) exercise. A spin echo EP sequence was employed with TE = 30 ms, TR = 4000 ms, FOV = 20 x 40 cm, 64 x 128 matrix. The changes in proton transverse relaxation rate (deltaR2, [s(-1)]) relative to values obtained before exercise were calculated from individual images at different times during and after exercise. RESULTS: At both 70 and 25% of MVC, the maximum deltaR2 after dynamic exercise (-8.38+/-0.32 s(-1) (70%), -6.47+/-1.23s(-1) (25%)) was significantly greater (P < or = 0.05) than after isometric exercise (-5.91+/-0.67 s(-1) (70%), -3.80+/-0.87s(-1) (25%)). Throughout the period that recovery was monitored, the recovery patterns of deltaR2 following isometric and dynamic exercise at both workloads remained parallel. CONCLUSIONS: We conclude that exercise-induced changes in MR images are influenced not only by workload and exercise duration but also by the type of exercise, and we postulate that these differences result from the different physiological responses elicited by the different types of exercise. PMID- 9741607 TI - Maximal oxygen uptake: "classical" versus "contemporary" viewpoints: a rebuttal. AB - Bassett and Howley contend that the 1996 J. B. Wolffe lecture is erroneous because: 1) A. V. Hill did establish the existence of the "plateau phenomenon," 2) the maximum oxygen consumption (VO2max) is limited by the development of anaerobiosis in the active muscle, and 3) endurance performance is also determined by skeletal muscle anaerobiosis because the VO2max is the best predictor of athletic ability. As a result, 4) cardiovascular and not skeletal muscle factors determine endurance performance. They further contend that Hill's "scientific hunches were correct," requiring "only relatively minor refinements" in the past 70 yr. But the evidence presented in this rebuttal shows that Hill neither sought nor believed in either the "plateau phenomenon" or the concept of the individual maximum oxygen consumption. These twin concepts were created by Taylor et al. (97) in 1955 and erroneously attributed to Hill. Rather Hill believed that there was a universal human VO2max of 4 L x min(-1). His error resulted from his incorrect belief that the real VO2 unmeasurable because it includes a large "anaerobic component," rose exponentially at running speeds greater than 13.2 km x h(-1). But Hill and his colleagues were indeed the first to realize the danger that a plateau in cardiac output (CO) and hence in VO2 would pose for the heart itself. For unlike skeletal muscle, the pumping capacity of the heart is both dependent on, but also the determinant of, its own blood supply. Thus, if the CO reaches a peak causing the "plateau phenomenon," the immediate cause of that peak will have been a plateau in myocardial oxygen delivery, causing a developing myocardial ischemia. The ischemia must worsen as exercise continues beyond the supposed VO2 "plateau." To accommodate this dilemma, Hill and his colleagues proposed a governor "either in the heart muscle or in the nervous system" necessary to prevent myocardial ischemia developing during maximal exercise. This governor would cause maximal exercise to terminate before the development of a plateau in either coronary flow, CO, or VO2, or the onset of skeletal muscle anaerobiosis. Accordingly, a new physiological model is proposed in which skeletal muscle recruitment is regulated by a central "governor" specifically to prevent the development of a progressive myocardial ischemia that would precede the development of skeletal muscle anaerobiosis during maximum exercise. As a result cardiovascular function "limits" maximum exercise capacity, probably as a result of a limiting myocardial oxygen delivery. The model is compatible with all the published findings of cardiovascular function during exercise in hypobaric hypoxia, in which there is a greater likelihood that myocardial hypoxia will develop. PMID- 9741608 TI - Biorhythmic influences on functional capacity of human muscle and physiological responses. AB - Previously, this laboratory has demonstrated that exhaustive aerobic exercise performance is not subject to significant chronobiological variation between 0800 and 2000 h, but certain physiological responses to maximal aerobic effort do fluctuate significantly within that time frame. PURPOSE: The purpose of the present investigation was to determine whether muscle performance, and selected physiological responses to resistance exercise, was significantly influenced by time of day effects. METHODS: Ten healthy, but untrained, men (21.1+/-0.6 yr, mean +/- SE) volunteered to participate in the study. In a balanced and randomized study design, each subject performed resistance exercise protocols on an isokinetic dynamometer with maximal effort at 0800 h, 1200 h, 1600 h, and 2000 h. Selected physiological and hormonal data were recorded before and immediately following the exercise stimulus. RESULTS: The data demonstrated significant chronobiological oscillation in peak torque, average power, maximal work in a single repetition, and total work per set. Interestingly, this oscillation was manifested only at the fastest velocities of limb movement utilized. Pre- and postexercise rectal temperature exhibited significant time of day fluctuation, as did postexercise blood pressure. Plasma levels of testosterone and cortisol also displayed significant biorhythmicity under both pre- and postexercise conditions. However, exercise-induced responses (pre- to postexercise differences) of those steroid hormones did not exhibit significant biorythmic variation. Although plasma concentrations of both testosterone and cortisol were highest at 0800 h, testosterone to cortisol ratios were greatest at 2000 h. CONCLUSIONS: In summary, these data suggest that maximal muscle performance does vary within the segment of the day when exercise typically occurs (0800-2000 h) but that this variation is specific to speed of movement. PMID- 9741609 TI - Lower intensity physical activity is advantageous for fat distribution and blood glucose among viscerally obese older adults. AB - OBJECTIVE: The influence of daily accumulated physical activity on blood glucose among older adults with varying obesity patterns is unknown. The purpose of this investigation was to determine if the blood glucose lowering effect of daily movement is modulated by distribution of adiposity in a community-based sample of older persons. METHODS: The study sample (N = 743) was mostly women (79.4%) with an average age of 74.5+/-0.3 yr. A question from the Yale Physical Activity Survey was the indicator of lower intensity physical activity. The response, answered in h x d(-1) spent in motion, was divided into tertiles (<3, 3 to <5, and > or = 5 h x d(-1)). Random blood glucose and total blood cholesterol were assessed via finger stick. The body mass index (BMI) and waist circumference (WC) delineated the categories of adiposity patterning as follows: nonobese (N = 354), BMI = 23.8+/-0.1 kg x m(-2) and WC = 80.3+/-0.4 cm; noncentral obese (N = 79), BMI = 30.8+/-0.1 kg x m(-2) and WC = 87.5+/-0.4 cm; and central obese (N = 310), BMI = 32.7+/-0.3 kg x m(-2) and WC = 103.3+/-0.5 cm. RESULTS: After adjusting for age, gender, race, medication use, and postprandial state, blood glucose levels were lower with greater amounts of reported daily movement in the centrally obese, 8.6+/-0.4 mmol x L(-1), 6.6+/-0.4 mmol x L(-1), and 6.3+/-0.4 mmol x L(-1) for <3 h x d(-1), 3 to <5 h x d(-1), and > or = 5 h x d(-1), respectively (P < 0.001). As the centrally obese increased their hours of moving about, their WC was observed to be less, 105.7+/-0.8 cm, 103.4+/-0.8 cm, and 102.9+/-1.0 cm, respectively, independent of age, gender, race, and medication use (P < 0.05). Neither blood glucose nor WC differed between categories of daily movement in the noncentral obese or nonobese. CONCLUSIONS: Our findings suggest that daily accumulated, lower intensity physical activity is advantageous for abdominal fat distribution and blood glucose among viscerally obese older adults. PMID- 9741610 TI - Exercise, smoking cessation, and short-term changes in serum lipids in women: a preliminary investigation. AB - PURPOSE: This study investigated the combined effects of exercise and smoking cessation on serum lipids. METHODS: Eighteen female smokers quit smoking using standard behavioral methods combined with exercise (N = 9) or with a nonexercise contact time control (N = 9). The smoking cessation program for both groups consisted of 12 weekly 1-h behavioral modification sessions held over 12 wk. Exercise training consisted of three supervised 45-min sessions per week for 12 wk. Contact control consisted of three health education lectures/discussions per week for 12 wk. Fitness (estimated VO2 peak), dietary variables, and fasting serum lipids and lipoproteins were assessed before and at the end of treatment. VO2 peak increased in the exercise subjects compared with the controls. RESULTS: Total caloric intake as well as total fat and carbohydrate increased significantly after smoking cessation in the controls, but there were no dietary changes in the exercise group. high density lipoprotein (HDL)-C2 increased (7.6 mg x dL(-1), P < 0.01) in the exercise group, whereas the increases in HDL and its subfractions did not attain statistical significance in the contact control group. Total cholesterol, low density lipoprotein (LDL)-C, and triglycerides did not change in either group. CONCLUSIONS: We conclude that exercise training magnifies the increase in HDL-C that usually occurs with smoking cessation. PMID- 9741611 TI - Cardiovascular and metabolic costs of forward, backward, and lateral motion. AB - PURPOSE: The purpose of this study was to compare the metabolic and cardiovascular responses of movement in forward (FM), backward (BM), and lateral (LM) directions. METHODS: Thirteen athletes with the following characteristics (mean +/- SD) were evaluated: age 21+/-3 yr, height 172.0+/-9.0 cm, weight 62.92+/-9.05 kg, and VO2max 54.13+/-7.42 mL x kg(-1) x min(-1). Subjects were evaluated at 80.45 and 134.08 m x min(-1). A repeated measures ANOVA was used for statistical analysis (P < 0.05). RESULTS: At 80.45 m x min(-1), the following respective VO2 mL x kg(-1) x min(-1) and heart rate (HR) beats x min(-1) responses were: FM = 12.42+/-2.29, 113+/-10; BM = 15.95+/-2.45, 132+/-16; and LM = 22.10+/-4.76, 140+/-15. Both VO2 and HR were significantly different between conditions: LM > BM > FM. At 134.08 m x min(-1), the following respective VO2 and HR responses were: FM = 27.15+/-2.51, 146+/-7; BM = 31.33+/-5.77, 168+/-11; and LM = 32.58+/-5.74, 169+/-10. At 134.08 m x min(-1) neither HR or VO2 were significantly different between LM or BM (LM, BM, > FM). Stride length and stride frequency were also significantly different between conditions. These results indicate the variation in the energy cost of FM, BM, and LM. PMID- 9741612 TI - Lactate distribution in the blood during steady-state exercise. AB - PURPOSE: The purpose of this investigation was to examine the plasma to red blood cell (RBC) lactate concentration ([La]) gradient and RBC:plasma [La] ratio during 30 min of steady-state cycle ergometer exercise at work rates below lactate threshold ( LT. Blood samples were taken from a heated forearm vein, immediately cooled to 4 degrees C in a dry-ice ethanol slurry, and centrifuged at 4 degrees C to separate plasma and RBCs. RESULTS: During >LT, plasma [La] rose to 8.8+/-1.1 mM after 10 min and remained above 6 mM. RBC [La] (4.9+/-0.7 mM) was significantly lower than plasma [La] at 10 min and remained lower throughout exercise. As a result, there was a sizable [La] gradient (approximately 3.5 mM) from plasma to RBC during most of >LT. In LT, the ratio of RBC [La]:plasma [La] was the same for both (0.58+/-0.02) and not significantly different from rest. CONCLUSIONS: These results refuted our hypothesis that the RBC:plasma [La] ratio would decrease at the onset of >LT exercise because of muscle lactate release exceeding the ability of RBCs to take up the lactate. Instead, there appears to be an equilibrium between plasma [La] and RBC [La] in arterialized venous blood from a resting muscle group as evidenced by the constant RBC [La]:plasma [La] ratio. PMID- 9741613 TI - Physical fitness, physical activity, and functional limitation in adults aged 40 and older. AB - PURPOSE: A cohort of middle-aged and older men and women were followed for an average of 5.5 yr to examine the association between physical fitness, physical activity, and the prevalence of functional limitation. METHODS: The participants received medical assessments between 1980 and 1988 and responded to a mail-back survey regarding functional status in 1990. RESULTS: Among 3495 men and 1175 women over 40 yr of age at baseline, 350 (7.5%) reported at least one functional limitation in daily or household activities at follow-up. The prevalence of functional limitation was higher among women than men. Physically fit and physically active participants reported less functional limitation than unfit or sedentary participants. After controlling for age and other risk factors, the prevalence of functional limitation was lower for both moderately fit (odds ratio = 0.4, 95% CI = 0.2-0.6) and high fit men (odds ratio = 0.3, 95% CI = 0.2-0.4), compared with low fit men. Corresponding figures for women were 0.5 (0.3-0.7) and 0.3 (0.2-0.5) for moderately fit and high fit women. The association between physical activity and functional limitation was similar to the data for physical fitness. CONCLUSIONS: These data support a protective effect of physical fitness and physical activity on functional limitation among older adults and extend this protective effect to middle-aged men and women. PMID- 9741614 TI - Longitudinal analysis of scaling VO2 for differences in body size during puberty: the Muscatine Study. AB - PURPOSE: The purpose of this study was to determine an appropriate method to "normalize" oxygen uptake (VO2) for body size in children and adolescents. METHODS: We examined allometric scaling factors for a cohort of 126 children (mean age at baseline = 10.3 yr) participating in a 5-yr follow-up study. Each year for 5 yr we measured peak VO2, submaximal VO2, body mass, height, body composition, and sexual maturation. We sorted the 5-yr data set by sexual maturation and gender and then used the generalized estimating equation method to estimate regression parameters that described the influence of log transformed body mass on log transformed VO2. All analyses were repeated using log transformed fat-free body mass (FFM) in lieu of log transformed body mass. RESULTS: Models using FFM appeared better at eliminating the effect of body size on VO2. In boys a univariate model with a FFM exponent of 0.91 and in girls a univariate model with a FFM exponent of 0.87 satisfactorily normalized peak VO2. However, we could not identify a common body size exponent for both boys and girls. CONCLUSIONS: Results support the use of allometric scaling of VO2 as a function of FFM for maturing boys and girls but indicate that the effects of maturation on the relationship between VO2 and body size differ between boys and girls. PMID- 9741615 TI - The effect of lifelong exercise on psychomotor reaction time: a study of 38 pairs of male monozygotic twins. AB - PURPOSE: The aim was to study the effect of lifetime physical activity on psychomotor speed. METHODS: Foot and dominant hand visual simple and choice psychomotor reaction times were studied among monozygotic twins (38 pairs) aged 35-69, discordant for lifetime exercise histories. RESULTS: There was a trend that some components of psychomotor reaction time were faster for frequent than for occasional exercisers, but the findings were not consistent for the hand and feet. After controlling for occupational physical activity, only choice decision time for the hand (26 ms, P < 0.01) and choice reaction time for the contralateral foot (51 ms, P < 0.05) both remained 7% faster. There was no trend for systematic differences in reaction times between twins engaged in regular exercise versus their siblings exercising infrequently. CONCLUSIONS: Results suggest a somewhat smaller effect of exercise than reported in previous studies. Reaction time may be significantly affected only by vigorous, frequent exercise. Thus, health promotion through exercise may be unlikely to have notable effects on reaction time. PMID- 9741616 TI - Variability in energy cost and walking gait during race walking in competitive race walkers. AB - PURPOSE: The aim of this study was to examine the variability of energy cost (Cw) and race walking gait after a 3-h walk at the competition pace in race walkers of the same performance level. METHODS: Nine competitive race walkers were studied. In the same week, after a first test of VO2max determination, each subject completed two submaximal treadmill walks (6 min length, 0% grade, 12 km X h(-1) speed) before and after a 3-h overground test completed at the individual competition speed of the race walker. During the two submaximal tests, subjects were filmed between the 2nd and the 4th min, and physiological parameters were recorded between the 4th and the 6th min. RESULTS: Results showed two trends. On the one hand, we observed a significant and systematic increase in energy cost of walking (mean deltaCw = 8.4%), whereas no variation in the gait kinematics prescribed by the rules of race walking was recorded. On the other hand, this increase in metabolic energy demand was accompanied by variations of different magnitude and direction of stride length, of the excursion of the heel and of the maximal ankle flexion at toe-off among the race walkers. CONCLUSION: These results indicated that competitive race walkers are able to maintain their walking gait with exercise duration apart from a systematic increase in energy cost. Moreover, in this form of locomotion the effect of fatigue on the gait variability seems to be an individual function of the race walk constraints and the constraints of the performer. PMID- 9741617 TI - Reliability of peak-lactate, heart rate, and plasma volume following the Wingate test. AB - PURPOSE: The 30-s Wingate Anaerobic Test (WAnT) has been used to assess anaerobic performance capacity and to evaluate physiological responses to supramaximal exercise. Blood lactate concentration ([La]) following supramaximal exercise is often used in the field and in the laboratory to assess the glycolytic contribution to exercise. Although the reliability of the performance in the WAnT has been established, this has not been the case with the WAnT's [La] response. Thus, the main purpose of this research was to study the test-retest reliability of peak [La] following the WAnT. Additionally, the test-retest reliability of the heart rate (HR) and plasma volume changes (deltaPV) response was also evaluated. METHODS: Twenty-nine subjects (15 male, 14 female) of diverse training levels as well as physical characteristics (mean +/- SD: 23.3+/-7.0 yr, 62.5+/-12.0 kg, 170.8+/-9.7 cm, and 16.3+/-6.2% fat) performed two WAnTs within 1 wk. Capillary blood was sampled from a prewarmed fingertip at rest, just before the WAnT and at 3, 5, 7, and 9 min following it. HR was also measured during these times. RESULTS: Mean-power (MP) (+/-SE) in test 1 and test 2 was 8.4+/-0.2 and 8.3+/-0.2 W X kg(-1) body mass, respectively. Peak [La] was attained 5-7 min following the WAnTs and was not significantly different between test 1 and test 2 (9.7+/-0.3 vs 9.8+/-0.3 mM, respectively). Peak HR occurred within 5 s post-WAnT and was not different between tests (170.8+/-2.2 and 171.3+/-2.2 beats X min(-1), in test 1 and test 2, respectively). Peak deltaPV was not different between tests (-12.0+/ 3.4 and -11.1+/-3.2%, in test 1 and test 2, respectively). The intraclass reliability coefficients for peak [La]. peak HR and deltaPV were 0.926, 0.941, and 0.878, respectively, whereas the corresponding value for MP was 0.982. CONCLUSIONS: We conclude that peak [La], peak HR, and deltaPV following the WAnT are reliable measures. PMID- 9741618 TI - Social life and the single nucleotide: foraging behavior in C. elegans. PMID- 9741619 TI - TATA box mimicry by TFIID: autoinhibition of pol II transcription. PMID- 9741620 TI - DNA damage and checkpoint pathways: molecular anatomy and interactions with repair. PMID- 9741621 TI - Crystal structure of a vertebrate smooth muscle myosin motor domain and its complex with the essential light chain: visualization of the pre-power stroke state. AB - The crystal structures of an expressed vertebrate smooth muscle myosin motor domain (MD) and a motor domain-essential light chain (ELC) complex (MDE), both with a transition state analog (MgADP x AIF4-) in the active site, have been determined to 2.9 A and 3.5 A resolution, respectively. The MDE structure with an ATP analog (MgADP x BeFx) was also determined to 3.6 A resolution. In all three structures, a domain of the C-terminal region, the "converter," is rotated approximately 70 degrees from that in nucleotide-free skeletal subfragment 1 (S1). We have found that the MDE-BeFx and MDE-AIF4- structures are almost identical, consistent with the fact that they both bind weakly to actin. A comparison of the lever arm positions in MDE-AIF4- and in nucleotide-free skeletal S1 shows that a potential displacement of approximately 10 nm can be achieved during the power stroke. PMID- 9741622 TI - Solution structure of a TBP-TAF(II)230 complex: protein mimicry of the minor groove surface of the TATA box unwound by TBP. AB - General transcription factor TFIID consists of TATA box-binding protein (TBP) and TBP-associated factors (TAF(II)s), which together play a central role in both positive and negative regulation of transcription. The N-terminal region of the 230 kDa Drosophila TAF(II) (dTAF(II)230) binds directly to TBP and inhibits TBP binding to the TATA box. We report here the solution structure of the complex formed by dTAF(II)230 N-terminal region (residues 11-77) and TBP. dTAF(II)230(11 77) comprises three alpha helices and a beta hairpin, forming a core that occupies the concave DNA-binding surface of TBP. The TBP-binding surface of dTAF(II)230 markedly resembles the minor groove surface of the partially unwound TATA box in the TBP-TATA complex. This protein mimicry of the TATA element surface provides the structural basis of the mechanism by which dTAF(II)230 negatively controls the TATA box-binding activity within the TFIID complex. PMID- 9741623 TI - Crystal structure of a Smad MH1 domain bound to DNA: insights on DNA binding in TGF-beta signaling. AB - The Smad family of proteins, which are frequently targeted by tumorigenic mutations in cancer, mediate TGF-beta signaling from cell membrane to nucleus. The crystal structure of a Smad3 MH1 domain bound to an optimal DNA sequence determined at 2.8 A resolution reveals a novel DNA-binding motif. In the crystals, base-specific DNA recognition is provided exclusively by a conserved 11 residue beta hairpin that is embedded in the major groove of DNA. A surface loop region, to which tumorigenic mutations map, has been identified as a functional surface important for Smad activity. This structure establishes a framework for understanding how Smad proteins may act in concert with other transcription factors in the regulation of TGF-beta-responsive genes. PMID- 9741624 TI - The DNA replication and damage checkpoint pathways induce transcription by inhibition of the Crt1 repressor. AB - We have identified the yeast CRT1 gene as an effector of the DNA damage and replication checkpoint pathway. CRT1 encodes a DNA-binding protein that recruits the general repressors Ssn6 and Tup1 to the promoters of damage-inducible genes. Derepression of the Crt1 regulon suppresses the lethality of mec1 and rad53 null alleles and is essential for cell viability during replicative stress. In response to DNA damage and replication blocks, Crt1 becomes hyperphosphorylated and no longer binds DNA, resulting in transcriptional induction. CRT1 is autoregulated and is itself induced by DNA damage, indicating the existence of a negative feedback pathway that facilitates return to the repressed state after elimination of damage. The inhibition of an autoregulatory repressor in response to DNA damage is a strategy conserved throughout prokaryotic and eukaryotic evolution. PMID- 9741625 TI - Cse4p is a component of the core centromere of Saccharomyces cerevisiae. AB - Histones are fundamental structural components of chromatin and are expected to play important roles in chromosome dynamics. Here, we present direct evidence that Cse4p, a histone H3 variant, is a structural component of the core centromere of S. cerevisiae. In histone H4 and Cse4p mutants, the core centromere chromatin structure is disrupted at restrictive temperature. Overexpression of Cse4p suppresses this defect in the H4 mutant, implying that the two proteins act together in centromere structure. We show by chromatin immunoprecipitation experiments that Cse4p is specifically cross-linked to centromeric DNA. Furthermore, by immunofluorescence microscopy, Cse4p is found in discrete foci consistent with that expected for centromeres. These results suggest the kinetochore is assembled on a specialized centromeric nucleosome containing Cse4p. PMID- 9741626 TI - A subcomplex of the proteasome regulatory particle required for ubiquitin conjugate degradation and related to the COP9-signalosome and eIF3. AB - The proteasome consists of a 20S proteolytic core particle (CP) and a 19S regulatory particle (RP), which selects ubiquitinated substrates for translocation into the CP. An eight-subunit subcomplex of the RP, the lid, can be dissociated from proteasomes prepared from a deletion mutant for Rpn10, an RP subunit. A second subcomplex, the base, contains all six proteasomal ATPases and links the RP to the CP. The base is sufficient to activate the CP for degradation of peptides or a nonubiquitinated protein, whereas the lid is required for ubiquitin-dependent degradation. By electron microscopy, the base and the lid correspond to the proximal and distal masses of the RP, respectively. The lid subunits share sequence motifs with components of the COP9/signalosome complex and eIF3, suggesting that these functionally diverse particles have a common evolutionary ancestry. PMID- 9741627 TI - A requirement for caveolin-1 and associated kinase Fyn in integrin signaling and anchorage-dependent cell growth. AB - Caveolin-1 functions as a membrane adaptor to link the integrin alpha subunit to the tyrosine kinase Fyn. Upon integrin ligation, Fyn is activated and binds, via its SH3 domain, to Shc. Shc is subsequently phosphorylated at tyrosine 317 and recruits Grb2. This sequence of events is necessary to couple integrins to the Ras-ERK pathway and promote cell cycle progression. These findings reveal an unexpected function of caveolin-1 and Fyn in integrin signaling and anchorage dependent cell growth. PMID- 9741628 TI - PGL-1, a predicted RNA-binding component of germ granules, is essential for fertility in C. elegans. AB - Germ cells are distinct from somatic cells in their immortality, totipotency, and ability to undergo meiosis. Candidates for components that guide the unique germline program are the distinctive granules observed in germ cells of many species. We show that a component of germ granules is essential for fertility in C. elegans and that its primary function is in germline proliferation. This role has been revealed by molecular and genetic analyses of pgl-1. PGL-1 is a predicted RNA-binding protein that is present on germ granules at all stages of development. Elimination of PGL-1 results in defective germ granules and sterility. Interestingly, PGL-1 function is required for fertility only at elevated temperatures, suggesting that germline development is inherently sensitive to temperature. PMID- 9741629 TI - Identification and disruption of a plant shaker-like outward channel involved in K+ release into the xylem sap. AB - SKOR, a K+ channel identified in Arabidopsis, displays the typical hydrophobic core of the Shaker channel superfamily, a cyclic nucleotide-binding domain, and an ankyrin domain. Expression in Xenopus oocytes identified SKOR as the first member of the Shaker family in plants to be endowed with outwardly rectifying properties. SKOR expression is localized in root stelar tissues. A knockout mutant shows both lower shoot K+ content and lower xylem sap K+ concentration, indicating that SKOR is involved in K+ release into the xylem sap toward the shoots. SKOR expression is strongly inhibited by the stress phytohormone abscisic acid, supporting the hypothesis that control of K+ translocation toward the shoots is part of the plant response to water stress. PMID- 9741630 TI - Specific synergy of multiple substrate-receptor interactions in platelet thrombus formation under flow. AB - We have used confocal videomicroscopy in real time to delineate the adhesive interactions supporting platelet thrombus formation on biologically relevant surfaces. Type I collagen fibrils exposed to flowing blood adsorb von Willebrand factor (vWF), to which platelets become initially tethered with continuous surface translocation mediated by the membrane glycoprotein Ib alpha. This step is essential at high wall shear rates to allow subsequent irreversible adhesion and thrombus growth mediated by the integrins alpha2beta1 and alpha(IIb)beta3. On subendothelial matrix, endogenous vWF and adsorbed plasma vWF synergistically initiate platelet recruitment, and alpha2beta1 remains key along with alpha(IIb)beta3 for normal thrombus development at all but low shear rates. Thus, hemodynamic forces and substrate characteristics define the platelet adhesion pathways leading to thrombogenesis. PMID- 9741631 TI - A novel adaptor protein orchestrates receptor patterning and cytoskeletal polarity in T-cell contacts. AB - Recognition of antigen by T cells requires the formation of a specialized junction between the T cell and the antigen-presenting cell. This junction is generated by the recruitment and the exclusion of specific proteins from the contact area. The mechanisms that regulate these events are unknown. Here we demonstrate that ligand engagement of the adhesion molecule, CD2, initiates a process of protein segregation, CD2 clustering, and cytoskeletal polarization. Although protein segregation was not dependent on the cytoplasmic domain of CD2, CD2 clustering and cytoskeletal polarization required an interaction of the CD2 cytoplasmic domain with a novel SH3-containing protein. This novel protein, called CD2AP, is likely to facilitate receptor patterning in the contact area by linking specific adhesion receptors to the cytoskeleton. PMID- 9741633 TI - Proceedings of the 1st Flaviviridae Symposium. PMID- 9741632 TI - Natural variation in a neuropeptide Y receptor homolog modifies social behavior and food response in C. elegans. AB - Natural isolates of C. elegans exhibit either solitary or social feeding behavior. Solitary foragers move slowly on a bacterial lawn and disperse across it, while social foragers move rapidly on bacteria and aggregate together. A loss of-function mutation in the npr-1 gene, which encodes a predicted G protein coupled receptor similar to neuropeptide Y receptors, causes a solitary strain to take on social behavior. Two isoforms of NPR-1 that differ at a single residue occur in the wild. One isoform, NPR-1 215F, is found exclusively in social strains, while the other isoform, NPR-1 215V, is found exclusively in solitary strains. An NPR-1 215V transgene can induce solitary feeding behavior in a wild social strain. Thus, isoforms of a putative neuropeptide receptor generate natural variation in C. elegans feeding behavior. PMID- 9741634 TI - Quasispecies and the implications for virus persistence and escape. AB - BACKGROUND: In the 1970s Manfred Eigen and colleagues proposed a new model of molecular evolution to explain adaptability and rapid evolution of simple replicons, as those that probably populated the earth at the onset of life. This model of evolution placed emphasis on mutant generation, to the point of invalidating the concept of wild-type genomes as a defined sequence of nucleotides. In striking similarity with the proposals for such early replicons, present-day RNA viruses consist of complex distributions of nonidentical but closely related genomes termed quasispecies. OBJECTIVES: To discuss indeterminations inherent to a quasispecies structure and to the analytical procedures to define it, biological implications of quasispecies, and the need to take into account this type of population structure, in order to design effective strategies to prevent and control diseases caused by highly variable viruses. RESULTS: Quasispecies have many biological implications, extending from viral pathogenesis to the emergence of new pathogens, rapid antigenic variation, and alterations in cell tropism, virulence, host range and viral gene expression. CONCLUSIONS: Diseases caused by highly variable RNA viruses prove very difficult to control and vaccine development against such viruses are largely unsuccessful. It is important to understand quasispecies composition and dynamics, as quasispecies are an important step in the natural history of RNA viruses. PMID- 9741635 TI - Natural history and molecular biology of hepatitis G virus/GB virus C. AB - BACKGROUND: The hepatitis G virus (HGV) or GB virus C (GBV-C) is a new member of the Flaviviridae family. The virus is transmitted by transfusion of blood, infusion of some blood products, and by parenteral exposure to blood during intravenous drug use (IVDU) and haemodialysis. Transmission from mother to infant and by sexual contact has also been documented. Although the virus has been found in patients with acute and chronic hepatitis, evidence of disease association has not been forthcoming. The majority of patients carry the virus in the absence of liver enzyme abnormalities. OBJECTIVES: To review what is currently known about HGV/GBV-C in order to evaluate its similarity with other members of the Flaviviridae and the association of the virus with disease. RESULTS: The genomic organisation of the virus is typical for Flaviviridae, with long 5' and 3' untranslated regions (UTR). However, a clearly identifiable nucleocapsid encoding region is lacking. Polyprotein synthesis is mediated through an internal ribosome entry site (IRES) contained within the 5' UTR. Phylogenetic tree analysis of sequences derived from this region has demonstrated the existence of at least three genotypes. Apart from serum, HGV-RNA has been detected in lymphocytes also, but the quasispecies present in the two compartments appear to be different. The envelope glycoprotein E2 lacks a hypervariable region and is potentially the target of a neutralising antibody response. CONCLUSION: Molecular analysis of HGV reveals close similarity of the virus with HCV. However, an association of the virus with liver disease remains unresolved and no association of the virus with hepatocellular carcinoma has been reported. PMID- 9741636 TI - Apoptotic cell death in response to dengue virus infection: the pathogenesis of dengue haemorrhagic fever revisited. AB - BACKGROUND: Dengue virus infection may be asymptomatic or lead to undifferentiated febrile illness or dengue haemorrhagic fever and dengue shock syndrome (DHF/DSS). The major clinical manifestations of DHF/DSS are high fever, haemorrhage, hepatomegaly and circulatory failure. OBJECTIVES: The relatively high level of viraemia only a few days after infection may reflect a large number of replication sites. However, the degree of cell injury in fatal cases of DHF/DSS is not sufficient to explain death and suggests metabolic disturbance rather than tissue destruction. This theory was investigated in this study. RESULTS: We demonstrated that replication of dengue virus in infected cells induces stress leading to apoptotic cell death in vitro and in vivo. CONCLUSIONS: The elimination of apoptotic bodies by phagocytic cells is a previously unsuspected pathway of dengue virus clearance from infected tissues. However, the mechanisms of host defence involving apoptosis and phagocytic cell activation may cause local tissue injury or transient homeostasis imbalance and may trigger further deleterious events. PMID- 9741637 TI - Pathogenesis of mucosal disease, a deadly disease of cattle caused by a pestivirus. AB - BACKGROUND: Two biotypes of pestiviruses, cytopathogenic (cp) and non cytopathogenic (noncp) viruses, are distinguished by their effects on tissue culture cells. In contrast to the bovine viral diarrhoea virus (BVDV) system, only a few cp border disease virus (BDV) and cp classical swine fever virus (CSFV) strains have been described. Antigenically closely related noncp and cp BVDV can be isolated from cattle with fatal mucosal disease (MD) and are called a virus pair. The generation of cp BVDV in an animal persistently infected with noncp BVDV is regarded as causative for the development of MD. OBJECTIVES: To analyse viral pairs of BVDV at the molecular level and thereby identify differences between the viruses of each pair. STUDY DESIGN: BVDV pairs were isolated from several animals coming down with MD; the genomes of the respective BVD viruses were sequenced on cDNA level. Studies concerning the polyprotein processing of each strain were carried out. RESULTS: Molecular analysis of BVDV pairs demonstrated a linkage between RNA recombination, generation of NS3 and the onset of fatal MD. CONCLUSION: The molecular analysis of BVDV pairs revealed that the respective cp strains arise by RNA recombination from noncp viruses. PMID- 9741638 TI - Immune mediated and inherited defences against flaviviruses. AB - BACKGROUND: Flavivirus infection elicits an abundant immune response in the host which is directed against a number of the viral proteins. Resistance to flavivirus-induced disease can also be controlled via a non-immune mechanism involving the product of a naturally occurring murine gene, Flv. OBJECTIVES: To review studies that have reported the mapping of epitopes on flavivirus proteins that elicit T- or B-cell immune responses in mice or humans and to discuss a possible mechanism for flavivirus-specific genetic resistance. STUDY DESIGN: Purified viral proteins and synthetic peptides were used to map B-cell epitopes. Purified proteins, vaccinia-expressed viral protein fragments and synthetic peptides were used to map T-cell epitopes. Congenic-resistant, C3H/RV and congenic susceptible, C3H/He mice and cell cultures were used to study the mechanism of genetic resistance to flavivirus infection. RESULTS: T- and B-cell epitopes have been mapped to the E, NS1 and NS3 proteins of several flaviviruses. Immune responses to the C, PreM, NS2a, NS4a, and NS5 proteins have also been documented. Data suggest that the Flv gene product acts intracellularly to suppress the synthesis of viral genomic RNA. CONCLUSIONS: Although flavivirus infection elicits an abundant immune response, this response is not always rapid enough to protect the host from developing encephalitis. During secondary infections both the humoral and cellular flavivirus-specific responses can confer protection. Dengue haemorrhagic fever (DHF) and dengue shock syndrome (DSS) appear to be caused by an overly vigorous immune response. In genetically resistant animals reduced production of virus results in a slower spread of the infection, which in turn allows time for the immune response to develop and to clear the infection before disease symptoms appear. PMID- 9741639 TI - Maternal recognition of foetal infection with bovine virus diarrhoea virus (BVDV) -the bovine pestivirus. AB - BACKGROUND: Pestiviruses are the veterinary viruses with genome homology to human hepatitis C virus (HCV). This group includes classical swine fever virus (CSFV), border disease virus of sheep (BDV) and bovine virus diarrhoea virus (BVDV). There are some similarities in the pathology of all three virus infections; in utero transmission to the foetus can cause early embryonic losses, severe congenital abnormalities and, particularly with BVDV, lifelong persistent infections. In situ hybridisation studies have demonstrated virus within reproductive tissues and the germinal centres of lymphoid tissue. OBJECTIVES: To examine the immune response of cattle throughout their pregnancy following infection with bovine pestivirus (BVDV) during the first trimester (before 110 days). STUDY DESIGN: In two experimental studies, heifers were infected with BVDV before 98 days gestation. Their antibody response was monitored during the remainder of the pregnancy. In another study, the antibody response of pregnant cattle was monitored following a natural infection of BVDV on a farm. Calves of the dams from all these three studies were examined, following birth, for persistent BVDV infection. RESULTS: It was observed that in dams carrying persistently infected foetuses, the immune response was markedly higher (13811 + 1273 U ELISA antibody) than in those dams carrying uninfected foetuses (2542+/ 588 U ELISA antibody). These results were used to establish an antibody threshold (10000 U ELISA antibody) to predict the virus status of unborn calves during a farm outbreak of BVDV infection. The combined results of experimental and farm studies showed that in dams with low antibodies, 5/15 calves were infected whereas in dams with high antibodies, 17/19 calves were infected. CONCLUSIONS: The predictive reliability of the assay appeared valuable but not secure. The ability of BVDV to infect the foetus with consequent maternal recognition, whilst remaining inaccessible to maternal immune exclusion, is a novel finding. PMID- 9741640 TI - The conformation of hepatitis C virus NS3 proteinase with and without NS4A: a structural basis for the activation of the enzyme by its cofactor. AB - BACKGROUND: Hepatitis C virus (HCV) NS3 proteinase activity is required for the release of HCV nonstructural proteins and is thus a potential antiviral target. The enzyme requires a protein cofactor NS4A, located downstream of NS3 on the polyprotein, for activation and efficient processing. OBJECTIVES: Comparison of the proteinase three-dimensional structure before and after NS4A binding should help to elucidate the mechanism of NS4A-dependent enzyme activation. STUDY DESIGN: We determined the crystal structure of NS3 proteinase of HCV BK isolate (genotype 1b; residues 1-189) and also the crystal structure of this proteinase complexed with HCV BK-NS4A (residues 21-34). RESULTS: The core region (residues 30-178) of the enzyme without cofactor (NS3P) or with bound cofactor (NS3P/4A) is folded into a trypsin-like conformation and the substrate P1 specificity pocket is essentially unchanged. However, the D1-E1 beta-loop shifts away from the cofactor binding site in NS3P/4A relative to NS3P, thereby accommodating NS4A. One result is that catalytic residues His-57 and Asp-81 move closer to Ser-139 and their sidechains adopt more 'traditional' (trypsin-like) orientation. The N terminus (residues 1-30), while extended in NS3P, is folded into an alpha-helix and beta-strand that cover the bound cofactor of NS3P/4A. A new substrate-binding surface is formed from both the refolded N-terminus and NS4A, potentially affecting substrate residues immediately downstream of the cleavage site. CONCLUSIONS: Direct comparison of the crystal structures of NS3P and NS3P/4A shows that the binding of NS4A improves the anchoring and orientation of the enzyme's catalytic triad. This is consistent with the enhancement of NS3P's weak residual activity upon NS4A binding. There is also significant refolding of the enzyme's N-terminus which provides new interactions with P'-side substrate residues. The binding surface for P'-side substrate residues, including the P1 specificity pocket, changes little after NS4A binding. In summary, we observe a structural basis for improved substrate turnover and affinity that follows complexation of NS3P with its NS4A cofactor. PMID- 9741641 TI - Repression of the PKR protein kinase by the hepatitis C virus NS5A protein: a potential mechanism of interferon resistance. AB - BACKGROUND: Chronic infection with hepatitis C virus (HCV) is associated with progressive liver damage, including the development of cirrhosis and hepatocellular carcinoma, and HCV is a leading cause of liver dysfunction worldwide. The current therapy for chronic HCV infection, interferon-alpha (IFN), is effective in a minority of HCV-infected patients. Several studies have demonstrated a correlation between therapeutic outcome and the amino acid sequence of a small region of the HCV non-structural 5A (NS5A) gene product. It has been suggested that this region, termed the interferon sensitivity determining region (ISDR), may mediate IFN resistance by directly interacting with one or more cellular proteins associated with the IFN-mediated antiviral response. OBJECTIVES: In an attempt to define the molecular mechanism by which the NS5A protein and the ISDR might contribute to HCV resistance to IFN, we examined whether NS5A could regulate the IFN-induced protein kinase, PKR, a primary mediator of the IFN-induced antiviral response. STUDY DESIGN: Multiple approaches, including in vitro assays using recombinant proteins, the transfection of recombinant clones into cultured cells, and in vivo studies in yeast, were used to examine the interaction of NS5A with PKR, as well as the functional significance of the interaction. An ISDR deletion mutant was prepared to evaluate the importance of the ISDR in mediating the NS5A-PKR interaction and the requirement of this region for PKR inhibition. RESULTS: NS5A repressed PKR activity through a direct interaction with the protein kinase catalytic domain. Both PKR repression and interaction required the presence of the ISDR. CONCLUSIONS: Inactivation of PKR may be one mechanism by which HCV avoids the antiviral effects of IFN. Thus,therapeutic strategies designed to block the NS5A PKR interaction may increase the efficacy of IFN therapy in HCV-infected individuals. PMID- 9741642 TI - Ribozyme gene therapy for hepatitis C virus infection. AB - BACKGROUND: The development of antiviral drugs for hepatitis C virus (HCV) infection represents a substantial challenge. Similar to human immunodeficiency virus (HIV), HCV is highly prone to mutation. It is, therefore, expected that potential HCV therapeutics currently under development, such as protease inhibitors, will suffer from the same shortcomings of HIV therapeutic drugs; the emergence of drug resistant viral mutants. Ribozymes (Rz) are enzymatic RNA molecules that can be engineered to specifically target any given RNA molecule. A therapeutic Rz can be manufactured and administered as a drug, or a Rz gene can be delivered and expressed intracellularly by gene therapy. For HCV therapeutics, we favour the gene therapy approach as delivery and in vivo expression of Rz genes will result in a constant and continuous supply of multiple intracellular Rz, offering less opportunity for the development of drug-resistant viral variants. OBJECTIVES: To utilise direct intravenous injection of hepatotropic viral vectors to transfer Rz genes directly into the hepatocytes of HCV-infected patients, resulting in degradation of the HCV positive strand RNA genome, the viral mRNAs, and even the negative strand RNA replication intermediate. We plan to circumvent the emergence of drug-resistant viral mutants by targeting multiple, highly conserved HCV RNA sequences simultaneously with multiple Rz genes expressed from a single vector. STUDY DESIGN: Rzs targeting conserved regions of the HCV positive and negative RNAs were transcribed in vitro and used to cleave HCV target RNAs. The most effective Rzs identified were then incorporated into adeno associated viral (AAV) vectors and adenoviral (AV) vectors and tested for their ability to inhibit HCV core expression in a tissue culture model. RESULTS: Several Rzs targeting highly conserved HCV sequences effectively degraded positive and negative strands of HCV RNA in vitro. Furthermore, substantial inhibition of HCV gene expression was observed in tissue culture using viral vectors to deliver and express Rz genes. CONCLUSIONS: Rz gene therapy has potential for the production of anti-viral drugs directed against HCV. Initial studies employing Rz gene therapy to produced anti-viral drugs against HCV have proved successful. Rz gene therapy may be a useful approach to overcome problems associated with anti-HCV drug design, such as the emergence of drug-resistant mutants. PMID- 9741643 TI - Evaluation of molecular strategies to develop a live dengue vaccine. AB - BACKGROUND: Millions of individuals are estimated to become infected with dengue virus each year, particularly in tropical and subtropical regions. Mortality is low but infection can lead to a severe form of dengue, characterised by haemorrhage and shock. A safe and effective vaccine against dengue is still not available. OBJECTIVE: To use the successful construction of dengue type 4 virus (DEN4) cDNA, which yields infectious RNA transcripts, to provide a new approach to the development of safe and effective dengue vaccines. STUDY DESIGN: The 3' and 5' noncoding (NC) regions of the genome were targeted to construct DEN4 deletion mutants, because the sequences in these regions are thought to play an important role in the regulation of viral replication. DEN4 cDNA was also employed to construct a viable chimeric virus with dengue type 1, 2 or 3 antigenicity, by substitution of heterotypic structural protein genes. RESULTS: Most viable mutants, recovered from the cDNA constructs, were partially restricted for growth in simian cells as analysed by plaque morphology assay and viral yield analysis. Several 3' NC deletion mutants which exhibited a range of growth restriction in cell culture were further evaluated for infectivity and immunogenicity in rhesus monkeys. Occurrence and duration of viraemia were reduced for these deletion mutants, compared to the wild type DEN4. Analysis of antibody response to infection in rhesus monkeys also indicated that some of these mutants were attenuated. These DEN4 deletion mutants represent promising live dengue vaccine candidates that merit further clinical evaluation. Chimera DEN1/DEN4 or DEN2/DEN4 which expresses DEN1 or DEN2 antigenicity were also used to infect monkeys. Most monkeys immunised with these chimeric viruses, singly or in combination, developed high titres of neutralising antibodies and were protected against homotypic wild type DEN1 or DEN2 challenge. CONCLUSIONS: DEN4 and its derived chimeric viruses of other three dengue serotype specificity, that contain appropriate attenuating mutations, have a potential use in a tetravalent live vaccine against dengue. PMID- 9741644 TI - Perspectives for a hepatitis C virus vaccine. AB - BACKGROUND: Natural hepatitis C virus (HCV) infection elicits poor immunity. Although HCV proteins elicit immune responses in virtually all cases of infection, the great majority of HCV infections become chronic. Currently, no vaccine is available for HCV despite an estimated incidence of approximately 50000 new cases per year in the USA alone. OBJECTIVES: To discuss how the problems associated with developing a vaccine against HCV infection may be overcome and describe recent progress made towards overcoming these problems and developing a vaccine. STUDY DESIGN: A cytofluorimetric assay that can assess the ability of a serum sample to neutralise the binding of the HCV-envelope glycoprotein E2 to human cells (neutralisation of binding or NOB assay) was developed. The assay was used to assess the levels of antibodies capable of neutralising E2 binding in the sera of vaccinated and carrier chimpanzees. RESULTS: Low titres of NOB antibodies were found in the majority of chimpanzees challenged with HCV infection. Chimpanzees immunised with the E1/E2 heterodimer developed NOB antibodies and high levels of neutralising antibodies. These chimpanzees were not protected from challenge with heterologous virus but were protected from subsequent chronic infection. CONCLUSIONS: A subunit vaccine composed of recombinant HCV proteins may protect from infection or chronic infection by different HCV genotypes. PMID- 9741646 TI - Francesco Bonaventura Cavalieri (1598-1647) PMID- 9741645 TI - Defective adenoviruses as novel vaccines for the Flaviviridae. AB - BACKGROUND: Vaccines against many flaviviruses, such as Japanese encephalitis virus (JEV), yellow fever virus (YFV) and tick-borne encephalitis virus (TBEV), have been successfully used for many years. Other diseases such as dengue fever (DF) and hepatitis C are still major public health problems as no licensed vaccines are in use. OBJECTIVES: To review studies on the use of defective recombinant adenoviruses (Rads) as experimental flavivirus vaccines and comment on their use to prevent infections with other members of the Flaviviridae such as hepatitis C virus. STUDY DESIGN: Recombinant adenoviruses, defective in their replication strategy, contain deletions in the E1 and E3 regions of the genome to increase the amount of foreign genetic material that can be inserted. The expression of foreign genes, inserted into these regions, can be driven by the adenovirus's own promoter, or by an additional viral promoters. CONCLUSIONS: Rads have been successfully used to raise protective immunity in experimental models of infection with several viruses. They can elicit both humoral and cell-mediated immunity and can be given parenterally or by oral administration. In addition, their hepatotropism makes them suitable for tackling diseases such as hepatitis C. Careful design of the vaccine vectors is advised to ensure their efficacy and safety, and as hepatitis C is a persistent infection, it may be advisable to design Rads containing genes encoding for non-structural proteins in preference to structural proteins. PMID- 9741647 TI - Artificial neural network and medicine. AB - The introduction of human brain functions such as perception and cognition into the computer has been made possible by the use of Artificial Neural Network (ANN). ANN are computer models inspired by the structure and behavior of neurons. Like the brain, ANN can recognize patterns, manage data and most significantly, learn. This learning ability, not seen in other computer models simulating human intelligence, constantly improves its functional accuracy as it keeps on performing. Experience is as important for an ANN as it is for man. It is being increasingly used to supplement and even (may be) replace experts, in medicine. However, there is still scope for improvement in some areas. Its ability to classify and interpret various forms of medical data comes as a helping hand to clinical decision making in both diagnosis and treatment. Treatment planning in medicine, radiotherapy, rehabilitation, etc. is being done using ANN. Morbidity and mortality prediction by ANN in different medical situations can be very helpful for hospital management. ANN has a promising future in fundamental research, medical education and surgical robotics. PMID- 9741648 TI - Anti-ulcer potential of flavonoids. PMID- 9741649 TI - Anti-implantation effect of a bone-marrow cytokine-BIM. AB - Successful implantation of blastocyst is dependent upon a cytokine induced localized immunosuppression at uterus. BIM, a bone marrow secreted bio immunomodulator (BIM) has been observed to have positive immunomodulatory activity in immunosuppressed cases. As pregnancy is associated with immunosuppression upregulation of the suppressed immune system by injection of BIM (conc. 0.08 microg/g b.wt once in rats b.wt. <150 gm or 0.2 microg/g b.wt thrice in rats weighing > 160 gm) is believed to prevent implantation. The anti implantation action of BIM is probably mediated via mononuclear cells at site of uterus, the effect is reversible and a single dose did not affect the estrous cycle. Multiple dose of BIM however, produce prolonged diestrous and this is probably an autonomic phenomena. PMID- 9741650 TI - Development of self-sustaining limbic status epilepticus by continuous ventral hippocampal stimulation followed by low dose pilocarpine in rats. AB - Sequential treatment of rats with low doses of lithium and pilocarpine, a high dose of pilocarpine, or continuous hippocampal stimulation [CHS] (9 epochs, 10 min each) is reported to result in status epilepticus (SE). We report a novel method to establish SE based on continuous ventral hippocampal stimulation (5 epochs) followed by low dose pilocarpine (40 mg/kg) challenge. Motor limbic seizures occured in all the control rats. The latency to spike activity was 15 +/ 1 min after pilocarpine administration. Ventral hippocampal [VHc] and cortical EEG recordings were used to monitor the protective effect of diazepam (5 mg/kg). Except phenobarbital, all the three drugs completely prevented all the phases of seizure activity. Initiation of spikes was significantly prolonged by phenobarbital pretreatment. Further study on the characteristics of these convulsions offers a unique possibility for the recognition of brain regions, pathways, and neurotransmitters engaged in the spread of seizures in this model. PMID- 9741651 TI - Effect of acute exercise on skin potential in sedentaries and trained athletes. AB - Endosomatic electrodermal activity (skin potential level and skin potential response) as an indirect indicator of sympathetic nervous system activity was measured in 35 sedentary male students and 22 trained athletes of two groups during resting and after an acute exercise. The aim of this study was to investigate the difference of skin potential parameters between sedentaries and trained athletes before and after the acute exercise in bicycle ergometer. In sedentaries' group while skin potential level (SPL) and latency showed no significant variations, skin potential response (SPR) decreased significantly after the exercise (P < 0.001). In athletes' group SPL increased (P < 0.01) and SPR decreased (P < 0.05) after the exercise but latency had no significant difference. In addition, athletes had significantly higher SPL and lower SPR values before and after the exercise comparing with the sedentaries. The increase of SPL in athletes' group was thought to depend on sweat duct pores which have been more active and open than sedentaries. Also the decrease in SPR in athletes' group was thought to depend on the lower sweating threshold in athletes. PMID- 9741653 TI - Additive anticonvulsant effect of flunarizine and sodium valproate on electroshock and chemoshock induced seizures in mice. AB - The efficacy of Flunarizine (FLU), a calcium channel blocker, in combination with conventional antiepileptic drugs, phenytoin (PHT), carbamazepine (CBZ), sodium valproate (VPA), and ethosuximide (ESM), at ED50 doses, were examined for protective effects against maximal electroshock seizures (MES) and pentylenetetrazol (PTZ) induced seizures in mice. In both models, only VPA and FLU showed significantly enhanced protection, which was additive ie. 100% protection. In the MES test, though FLU combined with PHT did show a slightly enhanced protection (66.6%), with CBZ there was no enhancement as compared to either drug alone. In the PTZ test, FLU with ESM showed 83% protection this however was not statistically significant. The findings of this study in mice suggest that FLU would be a suitable candidate for add-on therapy with VPA for clinical epilepsy. PMID- 9741652 TI - Preferential blockade by clozapine of hyperlocomotion induced by non-competitive NMDA antagonist MK-801. AB - Effect of clozapine on MK-801-induced hyperlocomotion and stereotypy as well as open field behavior was studied. Clozapine (0.1-7.5 mg/kg) dose-dependently blocked MK-801(0.5 mg/kg)-induced stereotypy. Both total and ambulatory responses were blocked by even the lower doses (0.1-0.5 mg/kg) of clozapine. In open field test, clozapine selectively blocked hyperambulation induced by MK-801 (0.1 mg/kg) whereas it potentiated MK-801 (0.1 mg/kg)-induced stereotypy at all the doses used. Haloperidol (0.25 and 0.5 mg/kg) and SCH 23390 (0.5 and 1 mg/kg) showed a dose-dependent effect on MK-801-induced behaviors while sulpiride (25 and 50 mg/kg) failed to modify MK-801-induced open field behavior. This study supports the preferential effect of clozapine on dopamine receptors located in mesolimbic area and further suggests the possibility of using open field behavior induced by MK-801 as a model for studying atypical antipsychotics. PMID- 9741654 TI - Dose response of sulphur mustard: behavioral and toxic signs in rats. AB - The present study elucidates the behavioral and toxic signs in rats following dermal application of sulphur mustard (SM). Graded doses of SM (0.10, 0.25, 0.50, 0.75 and 1.0 LD50) were topically applied to male Wister rats. The body weight as well as behavioral/toxic signs and symptoms were recorded at 1, 2, 3, and 4th day after application of SM. Sulphur mustard consistently decreased body weights of rats in a dose and time dependent manner with maximum decrease on 3rd day post treatment. Sedation and diarrhea were significant in response to doses of SM intoxication in rats. It is concluded that the body weight, sedation and diarrhea may be used as a reliable parameter in evaluating SM intoxication. It is also suggested that hydration and hypertonic saline must be used as a rescue agent within 1-3 days after exposure to SM. PMID- 9741655 TI - Comparative effects of hyoscine butylbromide and atropine sulphate on sleep architecture in healthy human volunteers. AB - The changes in sleep architecture, heart rate and respiratory rate to hyoscine butylbromide (HBB), a peripherally acting anticholinergic was studied. These effects were compared with that of atropine sulphate, a drug known to cross the blood brain barrier. The study followed a single blind cross over design with a one week washout period. Atropine sulphate (0.4 mg) and HBB (10 mg) were given intravenously to ten adult healthy male volunteers before sleep onset. Normal saline was used as control. All night sleep polysomnography was done with the standard montage for sleep staging. Respiration and airflow were also monitored. Rapid eye movement (REM) latency was significantly increased with both the drugs whereas the duration of REM sleep was decreased only with atropine. Slow wave sleep (SWS) was also increased significantly by atropine. There was no change in heart rate, or respiratory rate during any of the sleep stages. HBB affects the initiation of REM sleep whereas atropine affects both its initiation and maintenance. PMID- 9741656 TI - Hypoglycemic effect of Biophytum sensitivum in the alloxan diabetic rabbits. AB - Hypoglycaemic effect of a Neapalese plant Biophytum sensitivum was investigated in the alloxan diabetic male rabbits of different severities: subdiabetic (Alloxan recovered; AR), mild diabetic (MD) and severely diabetic (SD). Assessment of activity of the extract, prepared from the plant leaves, was done by fall in fasting plasma glucose (FPG) and improvement in the oral glucose tolerance test (OGTT), following single dose and prolonged administrations. Following single dose administration there was fall in 1 hour and 2.5 hour glucose values by 25.9% and 27.4% respectively in the subdiabetic rabbits, and by 36.9% and 37.7% in the mild diabetic rabbits. Improved GTT response is shown by fall in area under curve (AUC) from 16138 mg/dl to 12355 mg/dl (23.4%) in the subdiabetic rabbits, and from 19258 to 12238 mg/dl in the MD rabbits. More significant improvements occurred following one week of above treatment. The results prove that the plant material has significant hypoglycaemic effect, which is possibly due to pancreatic beta-cell stimulating action. To investigate its possible role in correction of other metabolic abnormalities of diabetes further long term studies are required. PMID- 9741657 TI - Tramadol, a centrally acting opioid: anticonvulsant effect against maximal electroshock seizure in mice. AB - The present study was designed to investigate the pro- or anticonvulsant effect of tramadol using maximal electroshock (MES) test. An attempt was also made to determine the possible opioid receptor mechanism involved. MES seizures were induced through transauricular electrodes (60 mA, 0.2s) and the seizure severity was assessed by the duration of tonic hindlimb extensor phase. Intraperitoneal (i.p.) administration of tramadol resulted in a dose-dependent anticonvulsant action; the ED50 for the effect was 33 mg/kg. The anti-MES effect of tramadol was antagonized by the low doses (0.05 and 0.1 mg/kg, s.c.) of MR 2266, a selective kappa receptor antagonist and also by the high doses (1.0 and 5.0 mg/kg, i.p.) but not the low doses (0.1 and 0.25 mg/kg) of naloxone. The results suggest that the anti-MES effect of tramadol is mediated by kappa receptors, since MR 2266 and naloxone (in high doses) are known to block these receptors. PMID- 9741658 TI - Pulmonary functions in Indian sportsmen playing different sports. AB - Regular exercise has proved to be beneficial for the human body and the lungs are no exception. The present study was undertaken to assess the relation between the quality of exercise performed and the quantitative effect of these exercises on the lungs. Pulmonary function tests of sportsmen engaged in various sports were compared with each other and with that of the controls. Players playing football (n=18), hockey (n=19), volleyball (n=20), swimming (n=20) and basketball (n=18) were chosen for this study. Medical students (n=20) were chosen as controls. The parameters taken into account in this study were forced vital capacity (FVC), forced expiratory volume (FEV-1), and peak expiratory flow rate (PEFR). The results indicate that all the sportspersons had a higher values of lung functions compared to the controls. Among the various groups of players chosen for this study, the swimmers showed the maximum increase in their lung functions. PMID- 9741659 TI - Effect of exercise on acid-base status and ventilatory kinetics. AB - The normal respiratory responses and changes in acid base status in twenty normal height, weight and age matched subjects were studied; using Auto Spiro AS 300 spirometer for ventilatory parameters and NOVA stat profile 3 analyser for gas analysis. Each subject performed a progressive incremental treadmill exercise by Bruce protocol to their symptom limited maximum. Minute ventilation (VE), tidal volume (VT) and frequency of respiration (f) increased significantly (P<0.001). Acidosis occured following exercise as pH of arterialized venous blood declined significantly (P<0.05). Gas analysis of arterialized venous blood showed a rise in pO2 (P<0.001) and a fall in pCO2 (P<0.001). Recovery of acid base status as well as gaseous pressure in blood did not occur after 10 min. Expired gas pCO2 declined significantly (P<0.05) and pO2 increase significantly (P<0.05). These pressures returned to resting levels 10 min after exercise. Thus in normal young adults heavy exercise caused an increment in ventilatory kinetics producing hyperpnoea which recovers after a rest of 10 min. Acidosis stimulates the respiratory centre to cause hyperventilation which tries to meet the added metabolic demands of strenuous exercise. PMID- 9741660 TI - Dicrotic notch in females. PMID- 9741662 TI - Haemodynamic responses to sciatic nerve stimulation during acute experimental anaemia in cats. PMID- 9741661 TI - Therapeutic potentials of fenugreek. PMID- 9741664 TI - Hemorheological parameters in coronary artery disease. AB - Hemorheological parameters are primary risk factors in ischemic heart disease (IHD). In the present study the relation of these parameters to the severity of coronary artery disease (CAD) was examined. The data of 109 patients (mean age: 55+/-9 years) undergoing coronary angiography and 59 healthy controls (mean age:35+/-10 years) were analyzed. Hemorheological parameters (hematocrit, fibrinogen level, plasma viscosity (PV) and apparent whole blood viscosity (WBV)) were determined and the circulatory index (CRI) was calculated. Patients were classified into three groups according to their coronary vessel state based on the coronary angiogram: Group 1 (n = 19, mean age: 53+/-8 years) without significant CAD, Group 2 (n = 19, mean age: 51+/-11 years) with single vessel disease, Group 3 (n = 71, mean age: 57+/-8 years) with multivessel disease. All the measured hemorheological parameters of IHD patients were significantly higher than those of controls. Fibrinogen and PV were significantly elevated in Groups 2 and 3 compared with Group 1 (p < 0.05 and 0.01). Hematocrit and WBV were significantly increased in Group 3 compared with Groups 1 and 2 (p < 0.05). CRI was significantly decreased in IHD patients, and it was also lower in Group 3 than in Group 2 (p < 0.05). These results indicate that hemorheological parameters may play a role in the pathogenesis and development of CAD. PMID- 9741663 TI - Erythrocyte hyperaggregation and thrombogenic dysfibrinogenemia. AB - Erythrocyte aggregation was measured in 12 patients with congenital dysfibrinogenemia. The results showed hyperaggregation in four patients who had presented a thrombotic disorder, while aggregation was entirely normal in patients with asymptomatic dysfibrinogenemia. None of the four symptomatic patients had any other anomaly of hemostasis, in particular no coagulation inhibitor deficit or anti-phospholipid antibodies. The possible involvement of erythrocyte hyperaggregation in the thrombotic process is discussed. PMID- 9741665 TI - Alterations in erythrocyte aggregability in diabetics: the influence of plasmatic fibrinogen and phospholipids of the red blood cell membrane. AB - In order to ascertain whether the increased aggregability observed in the red blood cells of diabetic patients is induced exclusively by plasma factors or is also influenced by membrane lipids, we examined the phospholipids of the erythrocyte membrane, the plasma fibrinogen concentration and erythrocyte aggregation in 86 insulin and non insulin-dependent diabetic patients. The data obtained show that the erythrocyte aggregability of the diabetic patients is higher than that of the control group (10.0+/-2.4 vs. 7.8+/-1.6%). This increased aggregability correlates not only with a higher fibrinogen concentration but also with changes observed in the membrane phospholipids. The percentage of sphingomyelin (SP) in the patients is higher than in the controls (22.6+/-6.8 vs. 18.4+/-5.4%) and that of phosphatidylserine (PS) is lower (9.5+/-6.1 vs. 12.1+/ 5.1%). No differences in the percentages of the other two phospholipids identified (phosphatidylcholine, PC, and phosphatidylethanolamine, PE) were observed. The increase in the saturated nature of the phospholipids of the erythrocyte membrane, which can be measured by the (SP + PC)/(PE + PS) ratio, is statistically related (r = 0.39; p < 0.05) to the increased red blood cell aggregability observed in these patients. PMID- 9741666 TI - The cholesterol/phospholipid ratio of the erythrocyte membrane in children with familial hypercholesterolemia. Its relationship with plasma lipids and red blood cell aggregability. AB - The cholesterol/phospholipid ratio (C/PL) of the red blood cell membrane, plasma lipids and erythrocyte aggregability were evaluated in 20 children with familial hypercholesterolemia (age: 10.4+/-4.6 years) but without detectable vascular injury. The results indicate that hypercholesterolemic children have a higher erythrocyte membrane C/PL ratio than the control group (0.81+/-0.23 vs. 0.65+/ 0.08, p < 0.01). This membrane lipid alteration correlates inversely with the plasma concentration of HDL-cholesterol (r = -0.558, p < 0.010). The patients also showed greater erythrocyte aggregability than the control group (8.21+/-1.11 vs. 6.25+/-1.24, p < 0.001), but this does not seem to correlate with the changes observed in the lipid composition of the cell membrane. These results suggest that from childhood, people with familial hyper-cholesterolemia show alterations in the lipid composition of the red blood cell membrane that are related to the changes observed in plasma lipids and appear prior to atherosclerotic vascular symptoms. PMID- 9741667 TI - Hemorheological, coagulative and fibrinolytic changes during autologous blood donation. AB - BACKGROUND: Clinical data suggest that autologous blood donation may prevent postsurgical venous thrombosis. If confirmed, this is probably due to beneficial effects in rheologic and hematologic variables which may be changed in patients as a result of repeated bleeding. STUDY DESIGN AND METHODS: To ascertain this point, we studied variations in hematological, hemorheological, coagulative and fibrinolytic parameters in 30 patients undergoing autologous blood donation. RESULTS: Whole blood viscosity (WBV), plasma viscosity and blood viscosity adjusted to 40% hematocrit, progressively and substantially decreased throughout the successive bleeding at all the shear rates considered. WBV was further reduced by presurgical hemodilution with autologous plasma which decreased the platelet and leukocyte count. The hemostasis and fibrinolysis variables, however, underwent no clinically significative changes. CONCLUSION: Repeated bleedings change most hemorheological variables. By decreasing cytocrit and viscosity, reducing aggregability and increasing blood cell deformability an optimal milieu to help prevent thrombosis is artificially created. PMID- 9741668 TI - Sequential analysis of the influence of blood storage on aggregation, deformability and shape parameters of erythrocytes. AB - The hemorheological and morphological changes in blood during storage under standardised conditions for 35 days were analysed by sequential determination of the variability of aggregation, deformability and shape of erythrocytes. The shape analysis was carried out by shape descriptors based on the projected area and perimeter, as measured by processing of erythrocyte images obtained from blood smears. The aggregation of erythrocytes was analysed from the data on the sequential variation of transmitted laser intensity after passing through the erythrocyte suspension. Similarly, the deformability of erythrocytes was determined in terms of passage time as obtained by analysis of flow of erythrocyte suspension through the cellulose membrane. The results show that the erythrocyte aggregation parameters and shape descriptors show significant variation, whereas the deformability is reduced (up to 5%) compared with that of the fresh samples. These variations may explain the observed changes in blood viscosity and viability of erythrocytes after their infusion in to the cardiovascular system. PMID- 9741669 TI - Kinetics of beneficial effect of pentoxifylline on persistent forms of arterial hypertension. AB - In some hypertensive patients a high level of arterial pressure proved resistant to the effect of Ca-antagonists. However, the addition of therapeutic doses of Pentoxifylline caused a significant decrease of blood pressure. The arterial pressure changes were found to be correlated with the index of erythrocyte aggregability (investigated using a highly sensitive "Georgian technique") in these patients. Thus, a pathogenetic link between blood pressure and hemorheological disorders could be conjectured. For a better understanding of the mechanism of these events the direct effect of Pentoxifylline on erythrocyte aggregability was investigated in vitro by using the blood samples of hypertensive patients possessing hemorheological disorders. The obtained results showed that the effect of Pentoxifylline (in therapeutic doses) was direct and that the dose-effect dependence was linear. From the obtained results we concluded that the beneficial effect of Pentoxifylline in hypertensive patients resistant to Ca-blockers is attained by eliminating the immediate cause of blood rheological disorders, the enhanced erythrocyte aggregability. PMID- 9741670 TI - A comparative study of the haemorheology of various breeds of dog. AB - Haemorheological parameters in nine breeds of dog were examined. Whole blood viscosity at both high and low shear rates differed significantly between the breeds with a 50% difference between the highest and lowest viscosity at high shear rate and a 140% difference at low shear rate. Athletic breeds (Greyhounds, Deerhounds) had the highest whole blood viscosities. Differences in viscosity correlated well with differences in haematocrit between breeds. When the blood samples were adjusted to a standard haematocrit (45%), there were no significant differences in viscosity. This implied that other rheological factors such as cellular deformability and plasma viscosity did not vary significantly between breeds, and direct measurement showed this to be the case. PMID- 9741671 TI - Photodecontamination of blood components: advantages and drawbacks. AB - Photochemical methods using photosensitizing photoactive drugs are very promising for blood product decontamination. Depending on the nature of virus - or parasite - bound photosensitizers, direct photochemical addition to virus components (DNA, proteins and lipids) occurs or the photosensitizer produces singlet oxygen inactivating viruses or parasites. The main advantage of this method is the lack of dark toxicity of presently used photosensitizers (psoralens, methylene blue, merocyanine 540, porphyrins/chlorins, phthalocyanines). In blood, the uptake of photosensitizers is not fully specific for infected cells. Therefore, normal cells and plasma proteins may suffer from the photodynamic action. Consequently, cell metabolism, rheological properties and surface markers may be altered and a slow loss of functionality occurs during storage. Data regarding pathogen inactivation in plasma and blood proteins, platelets and RBC concentrates are presented in comparison with the effect of photosensitization on normal blood components. Means for protecting normal components from photosensitization are also evaluated. PMID- 9741672 TI - Increased blood viscosity in iron-depleted elite athletes. AB - Since iron deficiency is associated with abnormal erythrocyte rheology, we investigated relationships between plasma ferritin and blood rheology in 36 male elite sportsmen (age: 22.38+/-0.9 years). On the whole, ferritin was negatively correlated with blood viscosity (r = -0.36, p < 0.05). When 23 subjects with low ferritin levels suggesting mild iron deficiency were compared with 13 matched sportsmen with normal ferritin levels, iron-deficient sportsmen were shown to have a higher blood viscosity at 1000 s(-l) (3.17+/-0.09 vs. 2.85+/-0.06 mPas, p < 0.05), explained by a higher plasma viscosity (1.38+/-0.02 vs. 1.31+/-0.02 mPa s, p < 0.05), while hematocrit and RBC rigidity index Tk were similar in the two groups. RBC aggregability index M (4.59+/-0.58 vs. 2.95+/-0.43 mPas, p < 0.05) and M1 (8.46+/-0.58 vs. 6.07+/-0.55, p < 0.01) were higher in iron-deficient subjects. Serum zinc was lower in iron-deficient sportsmen (0.73+/-0.02 vs. 0.83+/-0.02 mg/l, p < 0.01), but the score of early signs of overtraining was higher in this group (10.84+/-1.61 vs. 4.08+/-1.11, p < 0.01). These data suggest that mild iron deficiency as commonly seen in athletes, before anemia occurs, is associated with an increase in plasma viscosity and RBC aggregation, together with an increased subjective feeling of exercise overload. PMID- 9741673 TI - Xth European Conference on Clinical Hemorheology. Workshop Red Cell Aggregation, Lisbon, July 1, 1997. PMID- 9741674 TI - Negative correlation between plasma fibrinogen and insulin sensitivity measured with the minimal model technique. AB - We aimed at investigating relationship between plasma fibrinogen and insulin sensitivity, which are two major determinants of metabolic Syndrome X (insulin resistance syndrome). We designed a prospective study of 27 non-diabetic, non hypertensive subjects, presenting a wide range of body mass index BMI (10 men, 17 women; mean age+/-SEM: 35.9+/-2.2 years; BMI ranging from 21.1-45.2 kg/m2). Insulin sensitivity was assessed with the minimal model procedure, over a 180 min intravenous glucose tolerance test with iterative sampling. Fibrinogen levels were determined by the method of Clauss. The insulin sensitivity index SI (i.e., the slope of the dose-response relationship between insulin increased above baseline and glucose disposal) ranged from 0.0009 to 16 x 10(-4) min( 1)/(microU/ml), with a mean value of 4.76+/-0.73 x 10(-4). Mean values of plasma fibrinogen were 3.33+/-0.13 g/l, ranging from 2.21 to 5.07 g/l. There were highly significant negative correlations between SI and the level of plasma fibrinogen (r = -0.61, p = 0.0007) and between the basal effect of insulin BIE and plasma fibrinogen (r = -0.521, p = 0.005). Basal insulin was positively correlated to fibrinogen (r = 0.386, p = 0.046). When we analysed the data using partial correlation analysis, the negative relation between SI and fibrinogen was maintained independently from BMI (r = -0.45, p < 0.05). These data establish a strong negative association between insulin sensitivity and fibrinogen, involved in the increased cardiovascular risk of metabolic Syndrome X. PMID- 9741675 TI - Prenatal stress alters brain biogenic amine levels in primates. AB - In this study, we assessed behavioral responses to social separation at 8 months of age and cerebrospinal fluid (CSF) concentrations of biogenic amines and metabolites at 8 and 18 months of age in 12 rhesus monkeys derived from either stressed or undisturbed pregnancies. Compared to controls from undisturbed pregnancies, prenatal stress-derived monkeys had higher concentrations of 3 methoxy-4-hydroxyphenylglycol (MHPG), and 3,4-dihydroxyphenylacetic acid in CSF than controls. Norepinephrine and MHPG response to stress were both correlated between 8 and 18 months of age. There were few group differences in behavior during social separation; however, several behavioral differences between groups were found when monkeys were reunited with cage mates. Prenatally stressed monkeys spent more time clinging to their surrogates and exploring (including eating and drinking), while controls showed more locomotion and social play with their cage mates. Collectively, our findings suggest that chronic unpredictable psychological stress during pregnancy has long-lasting effects on noradrenergic and dopaminergic activity and behavior in the offspring of gestationally stressed primate mothers. PMID- 9741676 TI - Immediate imitation and joint attention in young children with autism. AB - There is growing scientific interest in the precursors to the ability of conceiving other people's minds. The present study investigates two candidate precursors, imitation and joint attention, in young children with autism and a control group of nonautistic children with a developmental delay. Children with autism were found to be impaired or delayed in both abilities. Gestural and procedural imitation were significantly related to mental age and chronological age in subjects with autism. Although the evidence for an autism-specific deficit appears to be stronger in the domain of joint-attention behaviors than it is in the domain of imitation, it seems premature to reject imitation as a possible precursor to the development of mindreading skills. Systematic investigations of the imitation deficit in autism are urgently needed. PMID- 9741677 TI - The implications of different developmental patterns of disruptive behavior problems for school adjustment. Conduct Problems Prevention Research Group. AB - Based upon developmental models of disruptive behavior problems, this study examined the hypothesis that the nature of a child's externalizing problems at home may be important in predicting the probability of and nature of school adjustment problems at school entry. Parent ratings were collected for a sample of 631 behaviorally disruptive children using the Child Behavior Checklist. Confirmatory factor analyses revealed differentiated ratings of oppositional, aggressive, and hyperactive/inattentive behaviors at home. Teacher and peer nominations assessed school adjustment at the end of first grade. As expected from a developmental perspective, aggressive behaviors indicated more severe dysfunction and were more likely to generalize to the school setting than were oppositional behaviors. Hyperactive/inattentive behaviors at home led to more classroom disruption than did aggressive or oppositional behaviors. Co-occurring patterns of oppositional/aggressive and hyperactive/inattentive behaviors were more common than were single-problem patterns, and were associated with broad dysfunction in the social and classroom contexts. The results were interpreted within a developmental framework, in which oppositional, aggressive, and hyperactive/inattentive behaviors may reflect distinct (as well as shared) developmental processes that have implications for the home-to-school generalization of behavior problems and subsequent school adjustment. PMID- 9741679 TI - Distorted perceptions in dyadic interactions of aggressive and nonaggressive boys: effects of prior expectations, context, and boys' age. AB - This study examined distorted self- and peer perceptions in aggressive and nonaggressive boys at preadolescent and early adolescent age levels. Subjects completed semantic differential ratings of themselves and of their peer partners following two brief dyadic discussion tasks with competitive inductions and a game-playing task with a cooperative induction. Subjects also rated their expectations for self- and peer behavior prior to the two competitive interaction tasks. Research assistants later rated videotapes of the interactions. Aggressive boys had more distorted perceptions of dyadic behavior as they overperceived aggression in their partners and underperceived their own aggressiveness. These distorted perceptions of aggression carried over for aggressive boys into the third interaction task with a cooperative induction, indicating these boys' difficulty in modulating these perceptions when the overt demand for conflict is no longer present in the situation. Results also indicated that aggressive boys' perceptions of their own behavior after the first interaction task is substantially affected by their prior expectations, in comparison to nonaggressive boys who rely more on their actual behavior to form their perceptions. PMID- 9741678 TI - Multiple risk factors in the development of externalizing behavior problems: group and individual differences. AB - The aim of this study was to test whether individual risk factors as well as the number of risk factors (cumulative risk) predicted children's externalizing behaviors over middle childhood. A sample of 466 European American and 100 African American boys and girls from a broad range of socioeconomic levels was followed from age 5 to 10 years. Twenty risk variables from four domains (child, sociocultural, parenting, and peer-related) were measured using in-home interviews at the beginning of the study, and annual assessments of externalizing behaviors were conducted. Consistent with past research, individual differences in externalizing behavior problems were stable over time and were related to individual risk factors as well as the number of risk factors present. Particular risks accounted for 36% to 45% of the variance, and the number of risks present (cumulative risk status) accounted for 19% to 32% of the variance, in externalizing outcomes. Cumulative risk was related to subsequent externalizing even after initial levels of externalizing had been statistically controlled. All four domains of risk variables made significant unique contributions to this statistical prediction, and there were multiple clusters of risks that led to similar outcomes. There was also evidence that this prediction was moderated by ethnic group status, most of the prediction of externalizing being found for European American children. However, this moderation effect varied depending on the predictor and outcome variables included in the model. PMID- 9741680 TI - Prenatal and perinatal influences on risk for psychopathology in childhood and adolescence. AB - The relationship between a range of prenatal and perinatal events and risk for psychopathology in offspring was examined. Prenatal and perinatal events investigated included maternal experiences, health, and substance use during pregnancy, obstetric complications, feeding practices, and infant health during the first year of life. Offspring diagnosis was based on structured interviews conducted with 579 adolescents on two occasions. Risk for later psychopathology was associated with a number of prenatal and perinatal factors. Major depression was associated with not being breast fed and maternal emotional problems during the pregnancy. Anxiety was chiefly associated with fever and illness during the first year of life and maternal history of miscarriage and stillbirth. Disruptive behavior disorder was associated with poor maternal emotional health during the pregnancy and birth complications. Risk for substance use disorder was associated with maternal use of substances during the pregnancy. Mediating effects of maternal depression, maternal-child conflict, and physical symptoms in the child, and moderating effects of gender of child and parental education were also evaluated. The limitations of this study are discussed and future research directions are suggested. PMID- 9741681 TI - Variables that initiate and maintain an early-onset trajectory for juvenile offending. AB - A trajectory defined by three time-ordered events was offered as a useful adjunct to building a development theory about antisocial behaviors. A sequence was defined with significant linkages between antisocial childhood behavior and early arrest and between early arrest and chronic offending. The majority of chronic offenders traveled through all three events in the sequence. Each event in the sequence shared a common process of disrupted family process plus frequent family transitions and marked social disadvantage. The findings support the hypothesis that the process that leads to antisocial behaviors at grade four may also maintain the entire sequence. The level of disrupted process at initiation and a time-based measure of involvement with deviant peers predicted which individuals moved forward in the sequence and which did not. The findings are consistent with the idea that the majority of chronic offending juveniles follow a trajectory that can be explained by a single theory. PMID- 9741682 TI - The co-occurrence of depression and substance abuse in late adolescence. AB - This article examines the role of adolescent social relationships in fostering the occurrence and co-occurrence of depression and substance abuse, using two waves of data from a community sample of adolescents (N = 900). Multinomial logistic response models were estimated to identify the extent to which risk and protective features of youths' family and peer relations were differentially linked with depressive symptoms, substance abuse, and their co-occurrence. Taking a within-person, configurational approach to adolescent adaptation, contrasts involved four subgroups of adolescents: those high on both depressed mood and substance abuse, those who experience neither problem, those evidencing high levels of depressive symptoms only, and those high on substance abuse only. Risk for depressive symptoms was differentiated by its association with conflict and lack of support in the friendship domain. Substance abuse was associated with negative peer pressure, but these youth were otherwise little different from youths with no problems. Whereas co-occurrence of depression and substance use was associated with more difficulties in both the family and peer environments, the most distinctive risk was that of low family support. Discussion centers on the developmental antecedents of co-occurring problems and family relations during adolescence. PMID- 9741683 TI - The role of child maltreatment and attachment style in adolescent relationship violence. AB - Utilizing attachment theory as a basis for conceptualizing close relationships among adolescents, this study investigated two important relationship risk factors (child maltreatment, and adolescent self-perceived insecure attachment style) as predictors of "offender" and "victim" experiences in youth relationships. In addition to considering the influence of these risk factors, we further considered their interaction in predicting conflict in close relationships. Of interest was the extent to which attachment styles may function as a moderator of the relationship between childhood abuse and current abuse in teen close relationships. High school students (N = 321) in grades 9 and 10 completed questionnaires tapping their histories of maltreatment, currently viewed styles of attachment, and conflict in close relationships over the past 6 months. Maltreatment alone emerged as the most consistent predictor, accounting for 13-18% of the variance in male's physically, sexually, and verbally abusive behaviors; in contrast, it was not highly predictive of female's abusive behaviors. Maltreatment was predictive of victimization experiences for both males and females. Attachment style did not substantially add to the prediction of relationship conflict beyond maltreatment; however, avoidant attachment style emerged repeatedly as a significant predictor of female abusiveness and victimization. Attachment self-ratings were found to function as a moderator of child maltreatment in predicting primarily male coercive behavior towards a relationship partner as well as predicting male's experience of coercion from a partner. Thus, the presence of childhood maltreatment and adolescent self perceived insecure attachment style applies predominantly to male youth. The implication of these gender differences for understanding relationship violence is discussed. PMID- 9741685 TI - Apolipoprotein E-deficient mice have impaired innate immune responses to Listeria monocytogenes in vivo. AB - Apolipoprotein E (apoE) influences both innate and acquired immunity in cultured cells. To determine whether apoE affects the immune system in vivo, Listeria monocytogenes (LM) was administered intraperitoneally (10(4) c.f.u.) to congenic C57BL/6 apoE-/- and +/+ mice (n = 12 in each group). Survival was assessed daily for 5 days. Deficiency of apoE significantly increased death by day 5 (P = 0.03). The majority of deaths occurred at day 4. Extent of infection after LM administration was assessed at day 3 by determining colony counts in hepatic and splenic extracts. ApoE+/+ mice had very low colony counts in both spleen and liver [mean +/- SE: 2.0 +/- 0.5 and 0.7 +/- 0.2 (x 10(4)), respectively, n = 8 in each group]; while apoE-/- mice had significantly increased counts in both spleen and liver [64 +/- 51 and 98 +/- 93 (x 10(4)), P = 0.05 and 0.03]. Serum concentrations of TNF-alpha were significantly increased in apoE-/- mice at day 3 compared to apoE+/+ mice (127 +/- 43 pg/ml versus 20 +/- 17, P = 0.003). LM induced more hepatic damage in apoE-/- mice compared to apoE+/+ mice as judged by increased serum concentrations of alanine aminotransferase at day 1 (apoE-/- 301 +/- 45 U/ml, apoE+/+ 101 +/- 9 U/ml, P = 0.01). The increased proliferation and mortality from LM in apoE-/- mice occurred prior to the initiation of acquired immune responses. Therefore, apoE-deficient mice have an impaired innate response to infection by LM. PMID- 9741684 TI - Human geranylgeranyl diphosphate synthase: isolation of the cDNA, chromosomal mapping and tissue expression. AB - We report the nucleotide sequence of human geranylgeranyl diphosphate (GGPP) synthase cDNA isolated from a fetal heart library. The 2.5 kb cDNA encodes a protein of 34 kDa. The protein contains six domains that have been identified previously in many other prenyltransferases. Recombinant, purified histidine tagged protein exhibited the enzymatic properties associated with GGPP synthase, namely the synthesis of GGPP from farnesyl diphosphate and isopentenyl diphosphate. Transient transfection of mammalian cells with a plasmid encoding the putative GGPP synthase resulted in a 55-fold increase in GGPP synthase activity. Taken together, these results establish that the cDNA encodes the mammalian GGPP synthase protein. The mRNA for GGPP synthase was expressed ubiquitously. Of the 16 human tissues examined, the highest expression of the mRNA was in testis. The mRNA levels in cultured HeLa cells were unaffected by alterations in cellular sterol levels and contrasted with the significant regulation of isopentenyl diphosphate synthase mRNA under these same conditions. Fluorescent in situ hybridization was used to map the single gene encoding human GGPP synthase to chromosome 1q43. PMID- 9741686 TI - Mucins and calcium phosphate precipitates additively stimulate cholesterol crystallization. AB - Human biliary mucin and calcium binding protein (CBP) influence formation of both calcium salt precipitates and cholesterol crystals and colocalize in the center of cholesterol gallstones. We investigated how physiological concentrations of these proteins regulate cholesterol crystallization in model biles, supersaturated with cholesterol and calcium salts, mimicking pathological human bile. Using polarizing light microscopy and nephelometry to assess cholesterol crystallization, the influence of calcium ions and calcium phosphate precipitates in the absence and presence of mucin, CBP, and human serum albumin was determined. Calcium phosphate precipitates stimulated cholesterol crystallization more strongly than soluble calcium. Mucin also stimulated, and with soluble calcium or calcium phosphate precipitates additively increased, the cholesterol crystal mass. In the absence of mucin, only human serum albumin plus CBP, not these proteins individually, decreased the stimulating effect of calcium phosphate precipitates but not of soluble calcium. However, seeding of calcium phosphate precipitates in biles with mucins resulted in near complete cholesterol crystallization within one day whether CBP and HSA were or were not also present. In conclusion, calcium salt precipitates plus human biliary mucins induce rapid and complete crystallization of cholesterol from model biles, little influenced by human biliary calcium binding proteins. PMID- 9741687 TI - Apolipoprotein B signal peptide and apolipoprotein E genotypes as determinants of the hepatic secretion of VLDL apoB in obese men. AB - We aimed to examine the effect of genetic polymorphisms of apolipoprotein B-100 (apoB) signal peptide and apolipoprotein E (apoE) on the hepatic secretion of very low density lipoprotein (VLDL) apoB in 29 men with visceral obesity. We studied apoB secretion using a primed (1 mg/kg), constant (1 mg/kg/h) intravenous infusion of [1-(13)C]leucine. The isotopic enrichment of VLDL apoB was determined using gas chromatography-mass spectrometry (GCMS). A multi-compartmental model was used to estimate the fractional turnover rate of VLDL apoB. Genotypes for the apoB signal peptide length polymorphism, 27 amino acid (SP27) and 24 amino acid (SP24), and apoE genotypes were determined using polymerase chain reaction. In subjects who were not apoE2 carriers and were homozygous for the SP27 of the apoB signal peptide, the hepatic secretion of VLDL apoB was significantly higher than in subjects who were not apoE2 carriers and were either heterozygous or homozygous for the SP24 allele (31.3 +/- 11.8 mg/kg fat-free mass/day, n = 8 vs. 16.9 +/- 12.2 mg/kg fat-free mass/day, n = 13, P = 0.01). In subjects who were not apoE4 carriers and were either heterozygous or homozygous for the apoB SP24 allele, the hepatic secretion of VLDL apoB was significantly lower than in subjects who were not apoE4 carriers and were homozygous for the SP27 allele (15.8 +/- 12.9 mg/kg fat-free mass/day, n = 13 vs. 27.4 +/- 11.5 mg/kg fat-free mass/day, n = 7, P = 0.03). The data suggest that in men with visceral obesity, the apoB signal peptide and apoE genotypes appear to be involved in the hepatic secretion of apoB. PMID- 9741688 TI - The hydrophobic surface of colipase influences lipase activity at an oil-water interface. AB - The interaction of pancreatic triglyceride lipase and colipase at an oil-water interface is required for efficient digestion of dietary fats and provides a model system for the interaction of proteins at biological membranes. Colipase has two important surfaces, a hydrophilic surface that interacts with lipase and a hydrophobic surface that presumably interacts with substrate. To begin our investigations into the role of the hydrophobic surface in the function of colipase, we replaced three neighboring tyrosine residues at positions 55, 58, and 59 in the hydrophobic surface with aspartic acid. Two of the three residues, Tyr55 and Tyr59, influenced the activity of colipase. Introducing aspartic acid at either position decreased the activity with long-chain triglycerides, but not with a short-chain triglyceride. Decreased ability of the mutants to anchor lipase to long-chain triglycerides did not explain the altered activity of the mutants. A mutant containing aspartic acid at positions 55 and 59 had no activity with any substrate and did not anchor lipase to either short- or long-chain triglycerides. These results identify the two tyrosine residues that interact with substrate and suggest that the hydrophobicity of the surface containing these tyrosines influences colipase function and the substrate selectivity of pancreatic triglyceride lipase. PMID- 9741689 TI - Degradation of blood group A glycolipid A-6-2 by normal and mutant human skin fibroblasts. AB - The degradation of blood group glycolipid A-6-2 (GalNAc(alpha1-->3)[Fuc alpha1- >2]Gal(beta1-->4)GlcNAc(beta1-->3)Gal(beta1-->4)Glc(beta1-->1')C er, IV2-alpha fucosyl-IV3-alpha-N-acetylgalactosaminylneolact otetraosylceramide), tritium labeled in its ceramide moiety, was studied in situ, in skin fibroblast cultures from normal controls, from patients with defects of lysosomal alpha-N acetylgalactosaminidase, and from patients with other lysosomal storage diseases. Uptake of the glycolipid with apolipoprotein E-coated liposomes was linear with time and with the amount of glycolipid added. In normal cells, the expected array of less polar products and some lipids resulting from re-using the liberated sphingosine, mainly sphingomyelin and phosphatidylcholine, were formed. In alpha N-acetylgalactosaminidase-deficient cells, the glycolipid was virtually not degraded; product formation was less than 2% of the normal control rate, suggesting that blood group A-active glycolipids contribute as storage compounds to the pathogenesis of this disease. The expected accumulation of degradation intermediates was seen in fucosidosis, and in Sandhoff, Gaucher, and Farber disease cells, whereas normal turnover rates were found in Tay-Sachs disease cells, G(M2) activator-deficient (variant AB of G(M2) gangliosidosis) and in sulfatide activator- (sap-B-) deficient cells. In G(M1) gangliosidosis and in sap precursor-deficient cells, the lysosomal glycolipid catabolism was found to be strongly retarded; accumulation of individual products could not be seen. Skin fibroblasts from patients with alpha-N-acetylgalactosaminidase deficiency (Schindler disease) cannot degrade the major blood group A glycolipid. PMID- 9741690 TI - Analysis of isoprenoid biosynthesis in peroxisomal-deficient Pex2 CHO cell lines. AB - ZR-78 and ZR-82 cells are two peroxisomal-deficient Chinese hamster ovary (CHO) cell mutants. These cells lack normal peroxisomes and show reduced levels of plasmalogen synthesis and other peroxisomal functions attributed to the deficiency of peroxisomal matrix enzymes. As we have recently identified two HMG CoA reductase proteins in CHO cells, a 97 kDa reductase localized in the ER and a 90 kDa reductase protein localized in peroxisomes, this enabled us to study the two reductase proteins for the first time in peroxisomal-deficient CHO cells. In this study we report the results of a detailed analysis of the isoprenoid biosynthetic pathway in the peroxisomal-deficient CHO cell lines ZR-78 and ZR-82. We demonstrate that total HMG-CoA reductase activity is significantly reduced in the peroxisomal-deficient cells as compared to the wild type cells. Analysis of the two reductase proteins in permeabilized cells indicated that in the ZR-78 and ZR-82 cells the 90 kDa peroxisomal reductase protein was mainly localized to the cytosol. We further show that the rates of both sterol (cholesterol) and non sterol (dolichols) biosynthesis were significantly lower in the peroxisomal deficient cells, when either [3H] acetate or [3H] mevalonate was used as substrate. In contrast, the rate of dolichol biosynthesis in the peroxisomal deficient cells was similar to that of the wild type cells when incubated with [3H] farnesol. In addition, we demonstrate that the peroxisomal-deficient cells exhibited increased rates of lanosterol biosynthesis as compared to wild type cells. The results of this study provide further evidence for the essential requirement of peroxisomes for cholesterol biosynthesis as well as for dolichol production. PMID- 9741691 TI - Transport and biotransformation of the new cytostatic complex cis diammineplatinum(II)-chlorocholylglycinate (Bamet-R2) by the rat liver. AB - Rat liver uptake and bile output of the cytostatic complex cis diammineplatinum(II)-chlorocholylglycinate (Bamet-R2) were studied. Up to 100 microM, Bamet-R2 uptake by rat hepatocytes in primary culture followed saturation kinetics (Vmax = 0.65 +/- 0.12 nmol/5 min per mg protein; K(M) = 45.2 +/- 10.7 microM). Bamet-R2 uptake was lower than that of cholylglycinate (CG) but higher than that of cisplatin. Replacement of 116 mM NaCl by 116 mM choline chloride did not significantly reduce Bamet-R2 uptake. Addition of 500 microM CG, cholic acid, estrone sulfate, or ouabain to 50 microM Bamet-R2-containing incubation media inhibited Bamet-R2 uptake. No liver biotransformation of Bamet-R2 occurred, as indicated by HPLC analysis of bile collected from anesthetized rats after intravenous administration of the drug. Bamet-R2 uptake and secretion into bile by isolated rat livers exceeded those of cisplatin but were lower than those of CG. Differences between Bamet-R2 and CG were more marked for bile output than for liver uptake. Thus, higher Bamet-R2 than CG or cisplatin liver content was found. Co-administration of Bamet-R2 and CG revealed that CG induced a slight reduction in Bamet-R2 uptake and a marked inhibition in Bamet-R2 bile output. By contrast, Bamet-R2 had no effect on CG on either liver uptake or bile output. In sum, the present data indicate that Bamet-R2 is efficiently taken up and secreted into bile by the rat liver by mechanisms shared in part by natural bile acids. PMID- 9741692 TI - LDL particle size and LDL and HDL cholesterol changes with dietary fat and cholesterol in healthy subjects. AB - We have conducted a dietary trial in 54 men and 51 women with a wide range of fasting cholesterol values to examine the use of low density lipoprotein (LDL) particle size to predict the lipoprotein response to dietary fat and cholesterol. After a 2-week low fat period, subjects were given two liquid supplements in addition to their low fat diet for 3 weeks each, one containing 31-40 g of fat and 650-845 mg of cholesterol, the other fat free. LDL particle type was determined by 3-15% gradient gel electrophoresis. On multiple regression, LDL type was independently related to plasma triglyceride (P < 0.001), waist circumference (P < 0.01), and high density lipoprotein (HDL) (P < 0.001) accounting for 56% of the variance in LDL type in the whole group. Change in LDL cholesterol with dietary fat and cholesterol was unrelated to LDL particle size in either men or women. However, change in HDL cholesterol in men was strongly related to LDL particle type (r = -0.52, P = 0.001) and change in HDL2 cholesterol in women was related to LDL particle type (r = -0.40, P < 0.01). In conclusion, we are unable to confirm the finding that LDL particle type can predict changes in LDL cholesterol following changes in dietary fat intake. However, LDL particle type can independently predict changes in HDL cholesterol in men and accounts for 27% of the variance. PMID- 9741693 TI - Accelerated lipoprotein uptake by transplantable hepatomas that express hepatic lipase. AB - To test the hypothesis that hepatic lipase plays a key role in lipoprotein removal in vivo, a novel system was used. Hepatoma cells (HTC 7288c) were transfected with a cDNA encoding hepatic lipase in culture and grown as solid tumors in vivo. In culture, transfected cells degraded chylomicron remnants and low density lipoprotein (LDL) somewhat more efficiently than untransfected cells. Tumors from the transplanted cells produced hepatic lipase localized to the surface of tumors from transfected cells but not tumors from non-transfected cells, grown in the same rat. The tumors from transfected cells removed, per gm of tissue, 34% (P < 0.001) more 125I-labeled LDL than tumors from non-transfected cells in the same animal. The uptake of chylomicron remnants (by tumors from transfected cells) was also modestly enhanced (15 +/- 6%, P < 0.005). There were no differences in the uptake of 125I-labeled albumin or 125I-labeled asialoglycoprotein. Compared to the liver, the untransfected tumors took up 12%, and the transfected tumors took up about 18% as much LDL per gram of tissue. The uptake of chylomicron remnants compared to liver was far lower. Both types of tumors had about twice as much LDL receptor related protein as the liver. Wild type tumors had the highest level of LDL receptor, twice hepatic lipase-secreting tumors, and six times that of the liver. Using the novel approach of transfecting transplantable tumor cells with hepatic lipase, the ability of hepatic lipase to facilitate the removal of apoB-containing lipoproteins was demonstrated. The liver still removes low density lipoprotein and especially chylomicron remnants more rapidly than the tumors, suggesting factors in addition to hepatic lipase and LDL receptor level play a major role in hepatic lipoprotein removal. PMID- 9741694 TI - Astrocytes are mainly responsible for the polyunsaturated fatty acid enrichment in blood-brain barrier endothelial cells in vitro. AB - To determine the respective roles of endothelial cells from brain capillaries and astrocytes in the conversion of circulating 18:2n-6 and 18:3n-3 into 20:4n-6 and 22:6n-3, respectively, a coculture of the two cell types mimicking the in vivo blood-brain barrier was used. During the culture period, endothelial cells cultured on an insert were set above the medium of a Petri dish containing or not a stabilized culture of astrocytes. Five days after confluence, labeled 18:2n-6 and 18:3n-3 (10 microM each) were added to the endothelial cells and incubated for 48 h. Analogous experiments were also performed by using each cell type cultured alone in the culture device. The distribution of radioactivity in lipids and fatty acids was studied in all the compartments of the culture device. Endothelial cells cultured alone weakly converted the precursor fatty acids into 20:4n-6 and 22:6n-3. When endothelial cells were cocultured with astrocytes, their content of polyunsaturated fatty acids increased dramatically. This effect was associated with the uptake of polyunsaturated fatty acids from the lower medium (astrocyte medium). These fatty acids were released by astrocytes after they were synthesized from the precursor fatty acids that passed through the endothelial cell monolayer into the lower medium. Polyunsaturated fatty acids were released by astrocytes as unesterified fatty acids and as phospholipids (mainly phosphatidylcholine and lysophosphatidylcholine) even when the medium was devoid of serum. These results suggest that astrocytes could play a major role in the delivery of essential polyunsaturated fatty acids to the barrier itself and to the brain. PMID- 9741695 TI - Delta6- and delta5-desaturase activities in the human fetal liver: kinetic aspects. AB - Delta6- and delta5-desaturase activities were studied in human fetal liver microsomes obtained after legally approved therapeutic abortion. Enzyme activities were measured by a radiochemical method using reverse-phase high performance liquid chromatography (HPLC). Free and phospholipid fatty acids were assessed in each liver sample by a combination of thin-layer chromatography (TLC) and gas-liquid chromatography (GLC) procedures. The kinetic measurements showed higher delta6-desaturase activity for the n-3 series than for the n-6 series. Apparent Km of 6.5, 3.9, and 24.5 microM and Vm of 7.5, 9.1, and 24.4 pmol x min( 1) x mg(-1) were obtained, respectively, for 18:2n-6 delta6-, 20:3n-6 delta5-, and 18:3n-3 delta6-desaturases. Beyond 30, 20, and 60 microM of 18:2n-6, 20:3n-6, and 18:3n-3 concentration, respectively, the enzyme activity deviated from Michaelis-Menten kinetics, suggesting an inhibition by excess substrate which is unlikely to occur in vivo as endogenous substrate concentration is much lower. We observed a breakdown in linearity between desaturase activity and microsomal protein concentration beyond 4-5 mg microsomal protein, whatever the enzyme or substrate. Both this phenomenon and the inhibition due to excess substrate should be taken into account in the determination of delta6- and delta5-desaturase activities. Comparison of concentrations of the respective endogenous substrates and the kinetic constants of each enzyme suggested that the higher delta6 desaturase activity observed for the n-3 series than for the n-6 series is not physiologically relevant in human fetal liver. PMID- 9741696 TI - Marked reduction in bile acid synthesis in cholesterol 7alpha-hydroxylase deficient mice does not lead to diminished tissue cholesterol turnover or to hypercholesterolemia. AB - These studies used mice that were deficient in cholesterol 7alpha-hydroxylase to determine the effects of reduced bile acid synthesis on cholesterol homeostasis. In mice lacking this enzyme, bile acid synthesis was reduced from 8.3 to 3.4 micromol/day per 100 g body weight, the intestinal bile acid pool was decreased from 62.5 to 13.2 micromol/100 g bw, and the proportion of hyodeoxycholate, relative to cholate, in this pool was significantly increased. Associated with these changes, intestinal cholesterol absorption decreased from 37% to <1% while triacylglycerol absorption and animal weight gain remained essentially unaffected. The very low rate of cholesterol absorption could be corrected by feeding the mutant mice cholate, but not hyodeoxycholate. The reduction in sterol uptake across the intestine was associated with a 2-fold increase in cholesterol synthesis in the small bowel and liver and an increase in fecal neutral sterol excretion from 15.2 to 35.7 micromol/day per 100 g bw. The size of the cholesterol pools in the plasma, various organs and whole animal remained constant. Thus, under circumstances where the excretion of sterol as bile acids was markedly reduced, total cholesterol turnover actually increased from 164 to 239 mg/day per kg bw. This study demonstrates the complex interactions between bile acid and cholesterol metabolism and the dramatic effects of eliminating a single gene product; however, even though a major catabolic pathway was deleted, cholesterol balance across the animal was maintained. PMID- 9741697 TI - Loss of phosphoserine polar group asymmetry and inhibition of cholesterol transport in Jurkat cells treated with cholesterylphosphoserine. AB - Cholesterylphosphoserine (CPHS) is a synthetic ester of cholesterol showing immunosuppressive activity. In the present study, we have used the T cell line Jurkat to investigate its mechanism of action. CPHS incorporates into cells reaching a molar ratio of 0.23 and 3.9 with the total phospholipid and cholesterol content, without inducing necrosis or apoptosis. CPHS incorporation elicits a dose-dependent binding of fluorescein isothiocyanate-labeled annexin V, suggesting that the steroid distributes in the external leaflet of plasma membrane exposing the phosphoserine group to the external cell environment and inserting the steroid ring into the phospholipid bilayer. In agreement with a preferential steroid association with sphingolipids, CPHS is included in a Triton X-100-insoluble complex when mixed with sphingomyelin and cholesterol. CPHS incorporation inhibits the esterification of low density lipoprotein (LDL) derived cholesterol, producing a minor influence on the endogenous synthesis of cholesterol and on the acyl-CoA:cholesterol acyltransferase activity. In this effect, CPHS is as potent as progesterone (IC50 of 3.5 microM). It is concluded that the insertion of cholesterylphosphoserine (CPHS) in the Jurkat plasma membrane neutralizes the asymmetric distribution of the phosphoserine group and inhibits the movement of cholesterol to the endoplasmic reticulum. As CPHS is a negatively charged steroid, this last effect may be linked to the perturbation of sphingolipid/cholesterol-based microdomains, proposed to play a role in cholesterol trafficking. PMID- 9741698 TI - Resistance of chylomicron and VLDL remnants to post-heparin lipolysis in ApoE deficient mice: the role of apoE in lipoprotein lipase-mediated lipolysis in vivo and in vitro. AB - The interaction of lipoprotein lipase (LPL) with triglyceride-rich lipoproteins is governed by a number of factors, such as apolipoprotein (apo) C-II. The role of apoE in lipolysis is controversial. We made the unexpected observation that apoE-deficient mice were resistant to heparin-induced lipolysis; this study aims at examining the underlying mechanism for this observation. Compared to wild-type mice, apoE-deficient mice had significantly higher very low density lipoprotein (VLDL) and chylomicron remnant (VLDL/CMR) concentrations and moderately lower lipase activity (15.5 +/- 1.3 mU/ml vs. 22.9 +/- 2.5 mU/ml). Unlike in wild-type mice where the injection of heparin reduced total plasma triglycerides by 50% and VLDL/CMR triglycerides by over 95%, the injection of heparin into apoE-deficient mice did not significantly affect plasma lipids. Similarly, in vitro, purified human LPL (hLPL) almost completely hydrolyzed VLDL/CMR isolated from wild-type mice, but had no effect on VLDL/CMR from apoE-deficient mice. However, when the amount of apoE-deficient VLDL/CMR was reduced to an equivalent level as in wild type mice, LPL hydrolyzed 94% of VLDL/CMR triglycerides. In order to increase the ratio of LPL to VLDL/CMR in vivo, we injected an adenovirus containing the human LPL cDNA into apoE-deficient mice, which produced marked liver-specific overexpression of LPL and significant reduction of VLDL/CMR (93%) and total plasma triglyceride concentrations (87%). Thus, apoE is not required for LPL activity in vivo or in vitro. Under certain pathological conditions, such as severe hyperlipidemia, the LPL pathway may be saturated and efficient lipolysis can proceed only if the ratio of substrate particles to LPL is adjusted to a more normal range. PMID- 9741699 TI - Baculovirus-mediated expression of recombinant rat phosphatidylcholine transfer protein. AB - Phosphatidylcholine transfer protein catalyzes intermembrane transfer of phosphatidylcholines exclusive of all other phospholipid classes. Although postulated to participate in phosphatidylcholine biosynthesis and biliary trafficking in liver, the molecular basis underlying the substrate specificity of phosphatidylcholine transfer protein remains to be elucidated. Having demonstrated the inability of Escherichia coli to express recombinant phosphatidylcholine transfer protein, we infected Spodoptera frugiperda (Sf9) cells with recombinant baculovirus. When assayed in vitro, cytosol of recombinant but not control infected cells demonstrated high levels of intermembrane phosphatidylcholine transfer activity and no transfer activity for phosphatidylethanolamine. A two-step purification protocol in which 10 mg of cytosolic protein was subjected to anion exchange chromatography followed by hydroxylapatite chromatography yielded 0.1 mg active protein which was >92% pure. The identity of purified protein was confirmed by matrix-assisted laser desorption-ionization mass spectrometry and by amino acid sequencing. Based on the recovery of 30% of PC transfer activity after purification, we estimate that recombinant rat phosphatidylcholine transfer protein accounted for approximately 3-6% of cytosolic protein mass of infected cells. These results demonstrate the utility of baculovirus for expressing recombinant phosphatidylcholine transfer protein and should facilitate studies designed to elucidate the structural biology and physiological functions of this uniquely specific phospholipid transfer protein. PMID- 9741701 TI - Multiple interactions between insect lipoproteins and fat body cells: extracellular trapping and endocytic trafficking. AB - The binding and internalization of a circulating insect lipoprotein, high density lipophorin (HDLp), by insect fat body cells was studied at the electron microscopic level using ultrasmall gold-labeled HDLp and DiI-labeled HDLp, which were visualized by silver enhancement and diaminobenzidine photoconversion, respectively. Internalization of HDLp seems to conflict with the selective process by which the lipids are transported between HDLp and fat body cells. The pathway followed by the internalized lipoproteins was investigated. In addition, the localizations of HDLp in fat body cells of young and older adult locusts were compared because of the previously reported age-related differences in distribution of cell-associated and internalized HDLp. In the present study, internalized labeled HDLp was observed in early endosomes, late endosomes, and putative lysosomes. In older adults, these labeled structures were much less abundant than in young adults. Moreover, in these animals, the labeled endosomal/lysosomal vesicles were located close to the plasma membranes. A more intense labeling was observed in the extracellular matrix in older adults compared to young adults. In both developmental stages, an apparent accumulation of labeled HDLp was found in extracellular spaces. We propose that this entrapment of HDLp may be essential for selective lipid transport between HDLp and fat body cells. PMID- 9741700 TI - Transmission of two novel mutations in a pedigree with familial lecithin:cholesterol acyltransferase deficiency: structure-function relationships and studies in a compound heterozygous proband. AB - Two novel mutations were identified in a compound heterozygous male with lecithin:cholesterol acyltransferase (LCAT) deficiency. Exon sequence determination of the LCAT gene of the proband revealed two novel heterozygous mutations in exons one (C110T) and six (C991T) that predict non-conservative amino acid substitutions (Thr13Met and Pro307Ser, respectively). To assess the distinct functional impact of the separate mutant alleles, studies were conducted in the proband's 3-generation pedigree. The compound heterozygous proband had negligible HDL and severely reduced apolipoprotein A-I, LCAT mass, LCAT activity, and cholesterol esterification rate (CER). The proband's mother and two sisters were heterozygous for the Pro307Ser mutation and had low HDL, markedly reduced LCAT activity and CER, and the propensity for significant reductions in LCAT protein mass. The proband's father and two daughters were heterozygous for the Thr13Met mutation and also displayed low HDL, reduced LCAT activity and CER, and more modest decrements in LCAT mass. Mean LCAT specific activity was severely impaired in the compound heterozygous proband and was reduced by 50% in individuals heterozygous for either mutation, compared to wild type family members. It is also shown that the two mutations impair both catalytic activity and expression of the circulating protein. PMID- 9741702 TI - Use of cytosolic triacylglycerol hydrolysis products and of exogenous fatty acid for the synthesis of triacylglycerol secreted by cultured rat hepatocytes. AB - The fatty acyl moieties incorporated into triacylglycerol (TAG) secreted by rat hepatocytes are derived from diacylglycerol (DAG) that is synthesized de novo through the phosphatidate pathway or derived from endogenous, cytosolic TAG after hydrolysis to DAG and re-esterification. We have used a dual-labeling technique (overnight labeling of cell TAG with [3H]oleate, followed by 3 h incubation with [14C]oleate) to quantify the contributions of the two sources towards TAG secretion by cultured rat hepatocytes. A wide range of TAG secretion rates was achieved by short-term incubation of the cells under a variety of conditions. There was no correlation between the overall amount of exogenous 14C-labeled fatty acid metabolized and the rate of either [14C]- or [3H]TAG secretion. By contrast, there was a strong positive correlation between the fraction of newly synthesized [14C]TAG that was secreted (the fractional secretion rate, FSR) and the absolute rate of TAG secretion. This suggests that the partitioning of DAG between (re)synthesis of cytosolic TAG and synthesis of secreted TAG is an important locus for the control of the rate of TAG secretion. Comparison of the ratio: oxidation/TAG secretion for 3H- and 14C-labeled acyl moieties indicated that, for all conditions studied, approximately half the acyl moieties already esterified to the glyceroyl backbone within cytosolic TAG remain unavailable for oxidation when this pool of TAG is mobilized for the synthesis of secreted TAG. The data provide evidence that hydrolysis of cytosolic triacylglycerol (TAG) does not proceed fully to the constituent fatty acids and glycerol, but only to the level of diacylglycerol, followed by re-modelling of approximately half of its acyl chains, before re-esterification to form secretory TAG. PMID- 9741703 TI - Surface enhanced Raman spectroscopic monitor of P. acnes lipid hydrolysis in vitro. AB - Surface enhanced Raman spectroscopy (SERS) at a silver microelectrode was used to monitor bacterial hydrolysis of triglycerides in lipid mixtures that model sebaceous gland secretions. Mixtures of wax esters, squalene, triolein, and triisostearin were used as model skin secretions. The transformation was followed in vitro as changes in the SERS caused by hydrolysis of triglyceride to fatty acid. The fatty acid was adsorbed as its carboxylate, which is readily identified by the characteristic band at ca. 1395 cm(-1). Co-adsorption of propionate was also observed. The technique can also confirm the presence of bacteria by detection of short chain carboxylic acids released as products of fermentation during the growth of these cells. PMID- 9741704 TI - Three polymorphisms associated with low hepatic lipase activity are common in African Americans. AB - We have shown previously that a hepatic lipase allele (designated -514T) is common among African Americans and contributes to low hepatic lipase activity in this population. To identify other hepatic lipase alleles associated with low hepatic lipase activity in this population, the coding region and intron-exon boundaries of the hepatic lipase gene were sequenced in 20 African American men with low hepatic lipase activity. Two polymorphisms (N193S and L334F) were associated with low post-heparin plasma hepatic lipase activity and were much more common in African Americans than in whites. This finding, together with our previous data on the -514T allele, indicates that at least three different hepatic lipase polymorphisms associated with low hepatic lipase activity are common among African Americans. Analysis of hepatic lipase haplotypes revealed that 97% of African Americans have at least one hepatic lipase allele that is associated with low hepatic lipase activity. PMID- 9741706 TI - Can we justify spermatid microinjection for severe male factor infertility? AB - During 1995 and 1996 the first spermatid pregnancies were announced with both round spermatid (ROSI) and elongated spermatid (ELSI) injections. These publications were flanked by live births from ROSI in a number of animal species, with resulting offspring appearing normal, healthy and fertile. However, the live births in humans heralded a scientific and ethical debate on the clinical use of this technology; and in a number of countries nationwide moratoria prohibiting spermatid microinjection were enjoined. Concerns surrounded the biological condition of spermatids and clinical implications of utilizing an immature spermatozoon for conception. Nevertheless, case reports and a few scientific studies on human spermatid conception have been published in recent years, and further polemic on testicular histopathology and prognosis has ensued. This paper reviews the current arguments on the clinical use of ROSI and ELSI, and evaluates the biology of the main contributory components of a spermatozoon to the subsequent embryo, namely the genetic material, the microtubular organizing complex and the putative oocyte activating factor. We also consider the relevant testicular histopathology and likely outcome in the context of the current birth rate from ROSI and ICSI. We conclude by considering the way forward for infertile men who require this technology to become genetic fathers, and whether the time is now appropriate to consider clinical trials. PMID- 9741705 TI - Micro- and macro-consequences of ooplasmic injections of early haploid male gametes. AB - This review refers to the evolution of ooplasmic injections of round spermatid nuclei (ROSNI) or intact round spermatids (ROSI). Conclusions from their preliminary application in the hamster, rabbit, mouse and human are discussed. Criteria for identification of round spermatids and guidelines/quality control for application of ROSNI/ROSI techniques are emphasized. Although all the animal offspring and the human newborns delivered after ROSNI/ROSI are healthy, additional research efforts are necessary to confirm the safety of these procedures and improve their outcome. PMID- 9741707 TI - Oocyte maturation and embryonic failure. AB - Embryonic development is readily compromised by imperfections introduced during the process of oocyte maturation. We discuss the nature and causes of these imperfections, particularly in oocytes exposed to inappropriate hormonal regimes in vivo or to culture systems designed to induce the maturation of oocytes in vitro. The acquisition of developmental competence involves the synthesis and storage of a wide range of molecules during oocyte growth followed by the reprogramming and ordered utilization of these stored products during maturation, fertilization and early embryogenesis. The regulatory signals for these molecular changes are produced by the follicle cells in response to circulating levels of gonadotrophins; we report that some ovarian stimulation protocols distort these signals thereby disrupting molecular reprogramming of the oocyte and reducing subsequent developmental competence. The aspiration of immature oocytes from antral follicles followed by their maturation in vitro is a potential alternative to hormonal stimulation of patients in IVF treatment. Although relatively successful in a variety of animals, the production of fully viable human embryos by in-vitro maturation is still unsatisfactory despite the use of a wide variety of culture protocols. Our data suggests that the key to maturation and embryo viability in vitro resides in the follicle cell compartment rather than the oocyte. Because of rapid luteinization changes, follicle cells in culture probably fail to provide the maturing oocyte with the necessary ordered set of instructive signals and nutrients needed for the acquisition of developmental competence. Although much remains to be discovered about the nature, concentration and transmission of signals, nevertheless it is already clear that different steroids, matrix metalloproteinases and growth factors are involved in conferring viability on the maturing oocyte. Major improvements in the yield of viable embryos from in-vitro matured oocytes can be anticipated from a systematic analysis of somatic signals from the pre-ovulatory follicle. PMID- 9741708 TI - The cryopreservation of ovarian tissue as a strategy for preserving the fertility of cancer patients. AB - The cryopreservation of ovarian tissue is a promising new method for conserving the fecundity of young cancer patients from the sterilizing effects of chemotherapy and/or radiotherapy. In murine and ovine studies the orthotopic insertion of frozen-thawed ovarian grafts into sterilized hosts has resulted in the birth of healthy offspring. Examination of human ovarian tissue after cryopreservation has shown that substantial numbers of morphologically normal and viable primordial follicles survive freeze-thawing. To date however, there is no efficient procedure for using the frozen-banked tissue to restore fertility to patients with ovarian failure. Autografting at an orthotopic or heterotopic site has the greatest potential for success but there is concern that the technique may reintroduce malignant cells to patients in remission from disease. Follicle isolation from the thawed tissue and growth to maturity in vitro is a preferable option but at present the technique is in its infancy. This review presents past and present research in the field of ovarian tissue cryopreservation and explores the possible strategies by which frozen-banked tissue from cancer patients could be used to restore fertility. PMID- 9741709 TI - Indications for cryopreservation of ovarian tissue. AB - For patients who are planning to have chemotherapy, radiotherapy or to undergo bilateral oophorectomy, loss of ovarian function will result in premature ovarian menopause and loss of fertility. For these women, although there is no successful method for the cryopreservation of human oocytes, ovarian tissue cryobanking is proposed with a view to its autotransplantation at a later date or the isolation and in-vitro maturation of oocytes. Embryo preservation is indeed not an option for single women and even for married women because delaying treatment for at least 2 months of in-vitro fertilization cycles is inappropriate and life threatening. Following the success of animal experiments, there have been reports of ovarian cryopreservation for women having to receive chemotherapy and/or radiotherapy. We present four case reports of ovarian tissue cryobanking and review the consequences of chemotherapy and radiotherapy on gonadal function, as well as the indications for freezing ovarian tissue. PMID- 9741710 TI - Structure-function relationship of follicle-stimulating hormone and its receptor. AB - Follicle stimulating hormone (FSH) is one of the two pituitary gonadotrophins involved in the regulation of gonadal function. Structurally, this gonadotrophin is a heterodimer composed of two non-covalently associated subunits containing several heterogenous oligosaccharide residues which play an important role in both the in-vivo and in-vitro bioactivity of the hormone. Its cognate receptor, which belongs to the superfamily of the G protein-linked cell surface receptors, also displays a high degree of functional and molecular complexity. Studies employing monoclonal antibodies, synthetic peptides and/or site directed mutagenesis, have unveiled some of the multiple structural determinants involved in FSH and FSH receptor function and interaction. Despite their structural complexity, both molecules exhibit a high degree of plasticity and diversity that allows formation of distinct ligand-receptor complexes capable of selectively activating or deactivating a variety of signalling pathways. Knowledge and mapping of the structural determinants and functional epitopes for intra- and extracellular hormone action are of paramount importance not only for a better and more detailed understanding of the molecular basis of FSH action and FSH receptor function but also for the rational design of analogues with predicted properties and effects. PMID- 9741711 TI - Inhibins and activins in human ovulation, conception and pregnancy. AB - The activins and inhibins are glycoproteins that belong to the transforming growth factor-beta superfamily and, as such, have diverse effects at many stages during growth and development. Originally identified by their effects on follicle stimulating hormone in males and females, the recent development of specific and sensitive assays for this group of polypeptides has permitted the elucidation of their role in 'fine-tuning' the hypothalamic-pituitary-gonadal axis. This review article focuses on the roles of inhibin and activin in female reproductive physiology with reference to possible future clinical applications in the investigation of infertility and abnormal pregnancy. PMID- 9741712 TI - Hyperandrogenic anovulation (the polycystic ovary syndrome)--back to the ovary? AB - Hyperandrogenic anovulation is characterized by polycystic appearance of the ovaries, elevated free serum testosterone with decreased concentrations of serum sex hormone binding globulin, an increased ratio of luteinizing hormone to follicle stimulating hormone and varying degrees of insulin resistance. We hypothesize that this is the result of variably increased 'ovarian androgenic insulin responsiveness' acting in combination with body mass. Women who develop hyperandrogenism and anovulation when lean (having normal serum insulin concentrations) represent the more severely affected individuals, whereas those who can resume ovulation by losing weight (and lowering their elevated serum insulin) represent a milder form. The tendency of these women to develop ovarian hyperstimulation syndrome despite hypophyseal desensitization suggests a primary ovarian defect which is apparently more pronounced in lean subjects. The unique ability of surgical damage to the ovary to induce ovulation, raises the possibility that inflammatory-like tissue remodelling has a major role in rescuing follicles from androgen-induced atresia. Approaches that may facilitate the study of this possible mechanism may include examination of in-vitro perfused, post-surgery mammalian ovaries and the elucidation of signal transduction mechanism(s) of insulin in the ovary, with special reference to cells emanating from affected women. PMID- 9741713 TI - Impact of heavy metals on hormonal and immunological factors in women with repeated miscarriages. AB - In 111 women with repeated miscarriages, the urinary excretion of heavy metals was determined in a challenge test with the chelating agent 2,3-dimercaptopropane 1-sulphonic acid in addition to hormonal, chromosomal, immunological and uterine investigations. The heavy metal excretion was correlated to different immunological (natural killer cells, T cell subpopulations) and hormonal (progesterone, oestradiol, prolactin, thyroid stimulating hormone) parameters. We conclude that heavy metals seem to have a negative impact on ovarian as well as on pituitary function. The heavy metal-induced immunological changes may interfere with the physiological adaptation of the immune system to the state of pregnancy with the result of a miscarriage. The observed heavy metal-induced hormonal and immunological changes may be important factors in the pathogenesis of repeated miscarriages. PMID- 9741715 TI - Microchips: an overview. PMID- 9741716 TI - AVA awaits release of new standard for universal readers. PMID- 9741717 TI - Endocarditis caused by Pasteurella caballi in a horse. PMID- 9741718 TI - Cellophane banding for the gradual attenuation of single extrahepatic portosystemic shunts in eleven dogs. AB - OBJECTIVE: To evaluate the efficacy and short term effects of a cellophane banding technique for progressive attenuation of canine single extrahepatic portosystemic shunts. DESIGN: A prospective trial of 11 dogs with single congenital extrahepatic shunts. PROCEDURE: Rectal ammonia tolerance testing and routine biochemical tests were performed preoperatively on all dogs. In seven dogs, preoperative abdominal Doppler ultrasonography was also performed. Exploratory laparotomy revealed a single extrahepatic portocaval shunt in each animal, which was attenuated using a cellophane band with an internal diameter of 2 to 3 mm. The abdomen was closed routinely. Follow-up biochemical analysis and abdominal Doppler ultrasonography or splenoportography were performed postoperatively. RESULTS: The shunt was not amenable to total ligation in 11 dogs, based upon reported criteria. All dogs recovered uneventfully from surgery without evidence of portal hypertension, and showed clinical improvement thereafter. Shunt occlusion was deemed to have occurred in 10 dogs based on resolution of biochemical and/or sonographic abnormalities. One dog continued to have sonographic evidence of portosystemic shunting when evaluated 3 weeks after surgery, despite normal ammonia tolerance, but was lost to subsequent follow-up. Two dogs, in which 3 mm cellophane bands were placed, experienced delayed shunt occlusion. CONCLUSION: Cellophane banding is simple to perform, and causes progressive attenuation of single extrahepatic shunts in dogs. Further work is needed to determine the maximum diameter of a cellophane band which will produce total attenuation, and the long-term safety and reliability of the treatment. PMID- 9741719 TI - Oesophageal impaction in a Canada goose (Branta canadensis). AB - An oesophageal impaction, consisting of plant material and nylon fishing line, and alimentary parasitism were diagnosed and treated in a Canada goose. At presentation the bird was non-ambulatory with flaccid neck muscles, lethargic, emaciated, dehydrated and had watery brown to green faeces. Palpation of the neck revealed a solid tubular mass ventrally in the mid-cervical region with gritty material cranial to it. Radiographs disclosed an oesophageal mass containing seed or grit-like radio-opaque material, and dilated cranial oesophagus containing radio-opaque material. Laboratory investigations revealed non-regenerative anaemia, heterophilia, lymphopenia, hypoproteinaemia, and many strongyle eggs in faeces. Treatment included supportive therapy, oesophageal gavage, oesophagotomy and drug therapy. The bird recovered and was released 27 days after initial presentation. PMID- 9741720 TI - Successful medical treatment of splenic abscesses in a horse. PMID- 9741721 TI - Enterotoxaemia in goats in Australia. PMID- 9741722 TI - What constitutes freedom from disease in livestock? PMID- 9741723 TI - Ovine Johne's disease: the risk associated with sheep imported into Western Australia. AB - OBJECTIVE: To assess the risk of Johne's disease not being detected in sheep imported from New South Wales into Western Australia. DESIGN: A stochastic simulation model. PROCEDURE: The process of importing sheep was broken down into steps and numbers or probabilities assigned to each. Controls on the movement of sheep included surveillance tests in source flocks and serological tests on sheep in consignments before and after transportation to Western Australia. The model calculated the risk of occurrence of Johne's disease in Western Australia and the success of the agar gel immunodiffusion test in identifying consignments with infected sheep. RESULTS: Negative surveillance tests in source flocks reduced the risk to about one twentieth of that when no surveillance tests were required. On average, Johne's disease was predicted to be introduced once in every 3 to 7 years when no testing of either the source flock or the sheep in consignments was required. When negative surveillance tests only were required the interval increased to once in every 63 to 111 years and, with the additional requirement that all sheep in each consignment must have a negative test before and after transport, the interval further increased to once in every 125 to 333 years. When only sheep in consignments were tested, the interval was calculated to be 8 to 14 years. CONCLUSION: A requirement that imports be derived from flocks which had negative surveillance tests to Johne's disease would provide significantly greater protection for the sheep industry in Western Australia. PMID- 9741724 TI - Survey of predation by domestic cats. AB - OBJECTIVES: To calculate the proportion of house cats which were observed by their owners to have caught prey and to describe the characteristics of these cats. DESIGN AND PROCEDURE: A telephone questionnaire was administered to a randomly selected population of 458 cat owners in metropolitan Perth. Specific questions were asked about demographic characteristics, habits and diets of the cats, and whether the owners had observed their cats catch prey in the 12 month period preceding the survey. RESULTS: The owners of 36% of 644 cats had observed their cats with prey in the 12 month period preceding the survey. Cats which spent more time outside, were neutered, cross-bred, originated from households with only one or two cats or were not fed meat were significantly more likely to be observed to predate. The body condition and diet (other than feeding meat) of cats did not influence the reported frequency of predation. CONCLUSION: Although cats are only one factor involved in the reduction in the numbers and diversity of Australian wildlife, restriction of the outside activities of cats is likely to diminish predation, particularly in areas close to native vegetation. PMID- 9741725 TI - Effects of delmadinone acetate on pituitary-adrenal function, glucose tolerance and growth hormone in male dogs. AB - OBJECTIVE: To characterise the effects of delmadinone acetate on the pituitary adrenal axis, glucose tolerance and growth hormone concentration in normal male dogs and dogs with benign prostatic hyperplasia. DESIGN: A prospective study involving nine normal male dogs and seven with prostatic hyperplasia. PROCEDURE: Delmadinone acetate was administered to six normal male dogs and seven dogs with benign prostatic hyperplasia at recommended dose rates (1.5 mg/kg subcutaneously at 0, 1 and 4 weeks). Three normal controls received saline at the same intervals. Blood concentrations of ACTH, cortisol, glucose, insulin and growth hormone were measured over 50 days. Intravenous glucose tolerance and ACTH response tests were performed before and after treatment in the nine normal animals. RESULTS: A substantial suppression of basal and 2 h post-ACTH plasma cortisol secretion was demonstrated after one dose in all dogs given delmadinone acetate. Individual responses after the second and third administration varied between recovery in adrenal responsiveness to continued suppression. Plasma ACTH concentration was also diminished after one treatment. No effects were evident on glucose tolerance or serum growth hormone concentrations. CONCLUSION: Delmadinone acetate causes adrenal suppression from inhibition of release of ACTH from the pituitary gland. Treated dogs may be at risk of developing signs of glucocorticoid insufficiency if subjected to stressful events during or after therapy. Neither glucose intolerance nor hypersomatotropism seems likely in male dogs given delmadinone acetate at the recommended dose rate, but the potential for excessive growth hormone secretion in treated bitches remains undetermined. PMID- 9741726 TI - A bone tumour resembling bizarre parosteal osteochondromatous proliferation in a wallaby. PMID- 9741728 TI - RSPCA responds to research claims. PMID- 9741727 TI - Chronic gypsum fertiliser ingestion as a significant contributor to a multifactorial cattle mortality. AB - OBJECTIVE: To assess the validity of claims that heavy metal contamination from an open-cut mine caused the death of 226 cattle on a nearby farm over a period of 18 months, and to investigate other possible contributing factors. PROCEDURE: A retrospective assessment of previous investigations combined with additional chemical analyses. RESULTS: Extensive chemical analyses produced no evidence of heavy metal contamination associated with the mine. Analysis of bones indicated exposure to fluoride in greater than normal amounts. The main source of fluoride seems to have been gypsum that was included in a feed supplement and also ingested from fertiliser dumps on paddocks. The gypsum itself may have contributed significantly to the ill health. Other factors probably affected some classes of animals, notably the young calves. CONCLUSIONS: What originally seemed to be a disease problem of single aetiology probably was an expression of interacting multifactorial causes. This investigation has highlighted the potential toxicity of gypsum to livestock and the need for further studies to establish its basis. PMID- 9741730 TI - Effect of nasal continuous positive airway pressure on pulmonary edema complicating acute myocardial infarction. AB - Cardiogenic pulmonary edema is a frequent cause of reparatory failure. We investigated the effects of nasal continuous positive airway pressure (CPAP) in patients with severe pulmonary edema associated with acute myocardial infarction. Twenty-nine consecutive patients were divided into 3 groups: firstly, 7 intubated patients who received mechanical ventilation at study entry comprised the intubation group. The rest of the patients were randomly assigned to either of the following 2 groups: 11 patients who received oxygen plus CPAP delivered by a nasal mask (CPAP group), and 11 patients who received oxygen only via face mask (oxygen group). All patients in the intubation group had cardiogenic shock. Two patients (18%) in the CPAP group and 8 patients (73%) in the oxygen group required mechanical ventilation with endotracheal intubation (p=0.03). The hospital mortality rate in the CPAP group (9%) was significantly lower than the oxygen group (64%, p=0.02). The pulmonary artery wedge pressure and heart rate were significantly lower in the CPAP group than in the oxygen group 24 h after study entry (p<0.05 and p<0.01). The mean pulmonary artery pressure 48 h after study entry was 18+/-5 mmHg in the CPAP group and 25+/-8 mmHg in the oxygen group (p<0.05). The PaO2/FiO2 ratio increased in the intubation group (168+/-69 to 240+/-57, p<0.05) and the CPAP group (137+/-17 to 253+/-67, p<0.01) 24 h after study entry. Arterial plasma endothelin-1 concentrations decreased significantly earlier in the CPAP group than in the oxygen group (p<0.05). In patients without cardiogenic shock, nasal CPAP lead to an early improvement in oxygenation and hemodynamics, and decreased the mortality rate. Early and active respiratory management is recommended in patients with pulmonary edema associated with acute myocardial infarction. PMID- 9741731 TI - Monitoring the local electrogram at the ablation site during radiofrequency application for common atrial flutter. AB - Recent studies have suggested that the attenuation of the local electrogram amplitude recorded from the ablation electrode during radiofrequency (RF) application predicts lesion growth. This study examined the time course of local electrogram amplitude during ongoing RF delivery in patients with common atrial flutter (AFl). In 71 patients with AFl. RF energy was applied to the anatomical isthmus. Termination of AFl was noted during 68 of 625 applications of RF energy. The changes in local atrial electrogram amplitude observed at all successful sites were analyzed. With increasing duration of the RF delivery, the electrogram amplitude decreased exponentially to reach a steady state within a mean duration of 17+/-3 sec, which was significantly longer than that of the steady-state temperature. The average decrease in the amplitude was 67+/-13%. In 16 patients in whom an increase in the power of RF energy had resulted in AFl termination, there was a dose-response relationship between the power and the amplitude decrease. The decrease in local electrogram amplitude appears to be a reliable marker for the efficacy of tissue heating and may be useful as an endpoint for individual applications. Local electrogram monitoring may offer an optimal energy strategy in AFl ablation. PMID- 9741732 TI - Efficacy of coronary artery bypass grafting in patients with a dilated left ventricle due to myocardial infarction. AB - This study was designed to clarify the efficacy of coronary artery bypass grafting (CABG) on left ventricular (LV) function in 16 patients with a dilated LV due to myocardial infarction (LV end-systolic volume index: LVESVI >60 ml/m2). All had attained complete revascularization. To estimate the LV wall motion quantitatively using echocardiography, a wall motion score (WMS) was used (LV was divided into 17 segments with a four-point scale: akinesis=3, severe hypokinesis=2, hypokinesis=1, normal=0 and then summed). Exercise stress tests were performed after surgery, revealing that anginal symptoms had vanished in all the patients. In 5 patients with a preoperative end-systolic volume index (ESVI) >100 ml/m2, the ejection fraction (EF) did not change, and both were under 30% (before to after: 26+/-4 to 26+/-4%). Neither the ESVI (148+/-50 to 133+/-39 ml/m2) nor the end-diastolic volume index (end-diastolic volume index (EDVI): 198+/-62 to 180+/-37 ml/m2) changed; the WMS did not change (33+/-2 to 33+/-3). During exercise, in spite of the increase in heart rate (HR) (at rest, 81+/-20; HR during exercise, 111+/-21 beats/min, p<0.005) and LV end-diastolic pressure (EDP) (22+/-9; 35+/-13 mmHg, p<0.02), both cardiac index (CI) (2.4+/-0.3; 2.6+/ 0.4 L/min x m2) and minute work (MW: 4.0+/-1.1; 4.1+/-0.4 kg x M/min) did not increase. In 11 patients with a preoperative ESVI <100 ml/m2, EF was extremely increased in 5 patients (more than 10%, 35+/-4 to 60+/-6%, p<0.005=improved subgroup) in whom the EDVI (130+/-16 to 120+/-13 ml/m2) did not change whereas the ESVI (82+/-14 to 48+/-7 ml/m2) was reduced. However, in the 6 remaining patients (ie nonimproved subgroup), neither ESVI (78+/-8 to 74+/-12 ml/m2), EDVI (115+/-10 to 115+/-20 ml/m2) nor EF (31+/-7 to 35+/-3%) changed. During exercise, HR (at rest, 88+/-13; during exercise, 108+/-11 beats/min, p<0.005), LVEDP (20+/ 6; 29+/-7 mmHg, p<0.01), CI (2.5+/-0.6; 3.3+/-0.5 L/min x m2, p<0.05), MW (4.6+/ 1.0; 6.5+/-1.5 kg x M/min, p<0.05) increased. The WMS in the nonimproved subgroup did not change (29+/-6 to 27+/-2), but in the improved subgroup it reduced after surgery (27+/-3 to 19+/-4, p<0.01). These data suggested that CABG in patients with a dilated LV was effective against anginal symptoms, but was restricted to left ventricular function. It may be possible to estimate postoperative LV function, including exercise tolerance, from the preoperative LVESVI. PMID- 9741733 TI - Efficacy and rebound phenomenon related to intermittent nitroglycerin therapy for the prevention of nitrate tolerance. AB - Intermittent transdermal therapy of nitroglycerin (NTG) has been recommended for the prevention of nitrate tolerance, but a rebound phenomenon has been reported to occur following removal of the NTG tape. The present study investigated the effects of intermittent NTG therapy on vasodilatory response and the intracellular production of cyclic GMP (cGMP). The study group comprised 12 healthy adults and measurements were taken of the platelet cGMP level, the venous volume (VV) (by forearm plethysmography) and the plasma levels of neurohormonal factors before and 5 min after administration of 0.3 mg of sublingual nitroglycerin (NTG) during the following 4 phases: (i) the control phase (8.00 h); (ii) the continuous phase (8.00 h; 7 days after continuous application of a 10 mg/24 h NTG tape); (iii) the intermittent application phase (8.00 h; 7 days after intermittent application of NTG tape, applied at 21.00 h and removed at 9.00 h); and (iv) the intermittent removal phase (13.00 h; 4 h after removal of the NTG tape in the intermittent phase). The percentage increase in cGMP (%cGMP) and venous volume (%VV) were significantly lower in the continuous phase than the control phase, but there was no difference between the control and the intermittent application phases. However, in the intermittent removal phase, the cGMP level before sublingual NTG, the %cGMP and the %VV were unchanged, but the VV before sublingual NTG was significantly lower than in the control phase. Plasma renin activity and the plasma level of angiotensin II were significantly increased in the continuous phase, the intermittent application phase, and the intermittent removal phase. In conclusion, intermittent transdermal NTG therapy prevented nitrate tolerance in the production of cGMP and vasodilation, but induced a rebound phenomenon after removal of the NTG tape. The rebound phenomenon following the tape removal may be related to some other mechanism, such as activation of neurohormonal factors. PMID- 9741734 TI - Venous response after lidocaine administration in arm veins. AB - We previously reported that cubital venous pressure (Pv) tended to increase initially, but this was followed by a drop in a dose-dependent response after intravenous lidocaine administration in subjects with various diseases. In this study we examined whether Pv responses after small-dose intravenous lidocaine administrations are related to the stimulating effect of lidocaine on vascular smooth muscle (VSM). In 5 subjects free of cardiovascular disease, Pv increased slightly with decreased pulsations after a 10 mg dose (p<0.01) with no change in central venous pressure. In the cinephlebographic test performed on 2 healthy volunteers, Pv increased during recovery from proximal venoconstriction caused by an injection of contrast medium mixed with 10 mg lidocaine. In 9 subjects with cardiovascular disease, deltaPv spread in the same directions (+ or -) after 5 and 10 mg drug administrations. In 6 of those tested with both drug doses, deltaPv had positive means and no significant difference was observed. Thus, Pv responses after small-dose lidocaine administrations are consistent with neither the stimulating effect of lidocaine nor with a dose-dependent response. They could be attributed to the spasmolytic effect of lidocaine on the basal tone of VSM, which could be modulated by disease conditions. PMID- 9741735 TI - The influence of changes in loading patterns on left ventricular relaxation in humans. AB - This clinical investigation was designed to determine the effect of changes in loading patterns on left ventricular (LV) relaxation when heart rate was maintained constant. Not only were changes noted in total load or time in which load is changed, but also the contour of the ascending aortic systolic pressure wave. Twenty patients were studied. LV and ascending aortic pressure were measured by a multisensor catheter under baseline conditions (C) and after an intravenous injection of 2.5 microg angiotensin (A) and sublingual administration of 0.3 mg nitroglycerin (N). A bipolar pacing catheter was placed in the right atrium to maintain a constant heart rate throughout the protocol. The augmentation index (AI), which characterizes the contour of the ascending aortic systolic pressure wave, was defined as the ratio of the height of the late systolic shoulder/peak to that of the early systolic shoulder/peak in the pulse. The rate of isovolumic LV pressure decline was calculated as a time constant (Tau). Ascending aortic systolic pressures (mmHg) were 127+/-29 (C), 158+/-20 (A) and 109+/-15 (N). AI were 1.61+/-1.14 (C), 2.08+/-1.11 (A) and 1.27+/-1.14 (N). Tau values (msec) were 49+/-4 (C), 54+/-4 (A) and 45+/-5 (N). Tau was prolonged proportionally with increasing AI (p<0.001, r=0.64). It was concluded that late systolic pressure augmentation in the ascending aorta is one important factor that influences the rate of isovolumic left ventricular pressure decline in humans. PMID- 9741736 TI - Detection of myocardial ischemia with a computer-assisted 12-lead 24-hour ECG monitoring system (EAGLE) in patients with suspected unstable angina. AB - This study was undertaken to evaluate the diagnostic value of a new device, the 'EAGLE' computer-assisted multiple-lead long-term electrocardiography (ECG) monitoring and analyzing system, in patients with suspected unstable angina, and to compare the results with the Holter monitor. A total of 101 patients with a history of suspected unstable angina underwent a simultaneous 24-h examination with the EAGLE and 2-channel Holter monitors. The diagnosis of unstable angina was established in 70 patients: 41 had significant organic stenosis, and 29 had coronary spasm. Ischemic ST deviations were detected 229 times in 44 patients (62.9%) with the EAGLE system and 101 times in 20 patients (28.6%) with the Holter monitor. The sensitivity of myocardial ischemia in unstable angina with the EAGLE system was significantly higher than that with Holter monitor (62.9 vs 28.6%, p<0.05). The difference of sensitivity was due mainly to the low detection rate of the Holter monitor for asymptomatic myocardial ischemia (EAGLE vs Holter; 187 times vs 72 times) and myocardial ischemia in the infero-posterior area in patients with organic stenosis (30 times vs none). It is concluded that the EAGLE system is a sensitive tool for the diagnosis of unstable angina in patients without significant ECG changes, and an excellent tool for evaluating silent myocardial ischemia or myocardial ischemia of the infero-posterior area. PMID- 9741737 TI - Quantitative assessment of myocardial 99mTc-sestamibi uptake during exercise: usefulness of response rate for assessing severity of coronary artery disease. AB - An increase of 99mTc-sestamibi uptake in the myocardium during exercise was defined as a response rate, and the feasibility of a response rate for detecting coronary artery disease (CAD) was tested. Eighty-seven patients with suspected CAD had myocardial perfusion imaging with 99mTc-sestamibi during exercise and at rest. A dose of 370 MBq of 99mTc-sestamibi was injected at the maximal level of exercise, and a myocardial image was obtained 90 min later (exercise image). Then, 740 MBq of 99mTc-sestamibi was administered at rest, and myocardial imaging was repeated (rest image). The exercise and rest images were corrected for physical decay and injected doses, and the exercise image was subtracted from the rest image to obtain the corrected rest image. A response rate was calculated as follows: (exercise image-corrected rest image)x100/corrected rest image (%). The global response rates of 20 patients without significant coronary stenosis (< or =50%) were higher than those of 67 patients with significant coronary stenosis (81+/-33% and 50+/-28%, p<0.01). Global response rates were correlated with the maximal rate pressure products during exercise (r=0.56, p<0.01) and delta rate pressure products (r=0.53, p<0.01). Regional response rates in myocardial areas perfused by stenotic coronary arteries of < or =50%, 75%, 90% and 99-100% were 60+/-24%,* 56+/-33%,* 40+/-23%* and 30+/-23%,* respectively, (*p<0.01 vs without significant coronary stenosis). The response rates decreased as the severity of coronary artery stenosis advanced, and distinguished between coronary stenoses of graded severity. Accordingly, the response rate from myocardial perfusion imaging with 99mTc-sestamibi may provide complementary information to the conventional inspection with myocardial tomography regarding the severity of CAD. PMID- 9741738 TI - Inhibitory effects of a subdepressor dose of L-158,809, an angiotensin II type 1 receptor antagonist, on cardiac hypertrophy and nephropathy via the activated human renin-angiotensin system in double transgenic mice with hypertension. AB - The effects of L-158,809, an angiotensin II type 1 receptor antagonist, on cardiac hypertrophy and nephropathy were examined using Tsukuba hypertensive mice (THM) carrying both human renin and angiotensinogen genes. Nine male THM aged 20 weeks were assigned to each of a no-dosage group and an L-158,809 dosage group, and L-158,809 was administered for 8 weeks. Nine age-matched male C57BL/6 mice were used as normal control animals. At 28 weeks of age, all of the mice were euthanized. Systolic blood pressure, urinary volume, water intake volume, urinary albumin excretion, heart weight and kidney weight to body weight ratios and a glomerulosclerosis index were measured. In the no-dosage group, the values of all of these parameters were larger than those in the control mice. In the L-158,809 group, all of the parameters showed significant improvement, except for blood pressure, which was not significantly different from that in the no-dosage group. These results suggest that the renin-angiotensin system played a crucial role in the cardiac hypertrophy and nephropathy in THM, and that L-158,809 exhibited strong curative effects on cardiac hypertrophy and nephropathy by blocking the angiotensin II type 1 receptor. PMID- 9741739 TI - Short-term and long-term inhibition of endogenous atrial natriuretic peptide in dogs with early-stage heart failure. AB - Early-stage heart failure (HF) is characterized by an increase in circulating atrial natriuretic peptide (ANP) without activation of the renin-angiotensin aldosterone system (RAAS) or body fluid retention. To test the hypothesis that elevated endogenous ANP suppresses the RAAS, maintains body fluid balance, and regulates vascular tone in early-stage HF, we assessed the effects of short-term and long-term inhibition of ANP on cardiorenal and neurohormonal functions. Short term antagonism was produced by bolus administration (3 mg/kg) of HS-142-1, an antagonist of guanylate-cyclase coupled ANP receptors, and long-term antagonism was produced by continuous infusion (1 mg/kg per h) of HS-142-1 for 8 h to dogs with early-stage HF induced by rapid ventricular pacing (270 beats/min, 8 days). In this experimentally produced HF, plasma ANP was significantly increased relative to the pre-pacing value, but not plasma renin activity (PRA) or plasma aldosterone level. HS-142-1 significantly suppressed plasma and urinary guanosine 3',5'-cyclic monophosphate (cGMP) levels, markers of endogenous ANP activity, in both experiments. Although mean arterial pressure and cardiac output did not change significantly, pulmonary capillary wedge pressure and right atrial pressure were elevated in both experiments. While short-term inhibition of ANP did not change PRA and aldosterone levels, long-term inhibition significantly increased these hormonal levels, resulting in decreases in urine flow rate, urinary sodium excretion rate, glomerular filtration rate, and renal plasma flow. These findings suggest that endogenous ANP plays a critical role in regulating venovascular tone, inhibiting activation of RAAS, and maintaining renal functions in early-stage HF. PMID- 9741741 TI - Goldenhar syndrome associated with various cardiovascular malformations. AB - This report presents a 54-year-old woman with Goldenhar syndrome featuring an epibulbar dermoid, left microtia and a left preauricular appendage, and synostosis of the vertebrae. Multiple cardiovascular malformations including Wolff-Parkinson-White syndrome, a partial anomalous pulmonary venous connection, patent ductus arteriosus, an anomalous origin of the coronary arteries, and a right-sided descending aorta were revealed by electrocardiography, echocardiography and cardiac catheterization. Goldenhar syndrome is very rare, but the frequency of cardiovascular malformations in this syndrome is 5-58%. It is necessary to perform a careful evaluation of general malformations, especially cardiovascular malformations. PMID- 9741740 TI - Mechanism for the cardioprotective effects of the calcium channel blocker clentiazem during ischemia and reperfusion. AB - To elucidate whether or not Ca channel blockers have an intrinsic benefit that cannot be attributed to the reduction of Ca2+ entry by pretreatment, time averaged intracellular Ca2+ concentration ([Ca2+]i) and energy-related phosphates were measured in isolated ferret hearts using nuclear magnetic resonance. In the drug-free ischemic group, [Ca2+]i increased significantly during 30 min of global ischemia at 30 degrees C and during 0-5 min of reperfusion. After 30 min of reperfusion, isovolumic left ventricular developed pressure recovered only to 63+/-7% of the pre-ischemic level (mean+/-SEM; N=5). Pretreatment with the Ca channel blocker clentiazem (10(-7) mol/L) itself depressed developed pressure by 53+/-9%. In the clentiazem group, [Ca2+]i showed no significant changes during ischemia or reperfusion. Recovery of developed pressure (87+/-8% of untreated level) was significantly higher than in the non-treated group (p<0.05). Nevertheless, when the negative inotropism of clentiazem was offset by increasing [Ca]o from 2 to 3 mmol/L, no beneficial effects of clentiazem were observed; [Ca2+]i increased significantly during 0-5 min of reperfusion, and developed pressure recovered only 60+/-7% of untreated level. These results indicate that reduction of Ca2+ entry from the extracellular space to the myocyte, as reflected by negative inotropism during pretreatment, is required for clentiazem to protect myocardium in a model of global ischemia and reperfusion. PMID- 9741742 TI - A case of hypertrophic obstructive cardiomyopathy with localized upper septal hypertrophy: reduction of left ventricular outflow obstruction by dual-chamber (DDD) pacing. AB - A patient with localized upper septal hypertrophy and medically uncontrolled severe outflow obstruction is described. His outflow obstruction was controlled by the implantation of a dual-chamber (DDD) permanent pacemaker. PMID- 9741743 TI - Mitochondrial encephalomyopathy (Kearns-Sayre syndrome) with complete atrioventricular block: a case report. AB - A pacemaker was implanted into a 17-year-old man with cardiac failure due to complete atrioventricular block complicated by mitochondrial encephalomyopathy (Kearns-Sayre syndrome). Due to the possible complication of latent myocardial dysfunction, it was decided to implant the dual chamber pacemaker (DDD) and the operation mode was set to DDD 70 ppm 1 year after implantation; this alleviated the cardiac failure. In this case, the necessity of preventive pacemaker implantation in the early stage of cardiac failure was recognized. PMID- 9741744 TI - Prosthetic ball valve endocarditis due to Gemella species. AB - A case is presented of endocarditis that was affecting a prosthetic ball valve (Starr-Edwards) and which was caused by Gemella species. A 57-year-old man was admitted with a 3-day history of abdominal pain with fever. At the time of admission, his temperature was 37.7 degrees C and laboratory tests showed elevated inflammatory parameters and an increased neutrophil count. However, transthoracic echocardiogram showed no vegetation. During hospitalization, Gemella spp. were detected by blood culture, and a transesophageal echocardiogram showed vegetation on the prosthetic valve. He was treated with intravenous ampicillin and astromycin, and also underwent valve replacement. This is the first case in Japan of infective endocarditis of a prosthetic valve due to Gemella spp. PMID- 9741746 TI - Kugelberg lecture: principles and pitfalls of nerve conduction studies. AB - Optimal application of the nerve conduction study depends on an understanding of the principles and a recognition of the pitfalls of the technique. The conventional methods deal primarily with distal nerve segments in an extremity. Other techniques allow one to assess nerve segments in less accessible anatomical regions, to improve the accuracy in precisely localizing a focal lesion, and to increase the sensitivity in detecting subclinical abnormalities. Despite certain limitations, nerve conduction studies can provide diagnostically pertinent information if they are used judiciously in the appropriate clinical contexts. PMID- 9741745 TI - Exercise-induced monomorphic ventricular tachycardia from the left ventricle outflow tract. AB - A case of exercise-induced idiopathic ventricular tachycardia (VT) arose from the left ventricular outflow tract. The QRS morphology of the VT was Rs pattern in V1 and R pattern in the lateral leads with inferior axis. The pacing at the superior interventricular septum near the mitral anulus produced the best pace mapping. Radiofrequency application to this site suppressed the VT. PMID- 9741747 TI - Dynamic functional coupling of high resolution EEG potentials related to unilateral internally triggered one-digit movements. AB - Between-electrode cross-covariances of delta (0-3 Hz)- and theta (4-7 Hz) filtered high resolution EEG potentials related to preparation, initiation. and execution of human unilateral internally triggered one-digit movements were computed to investigate statistical dynamic coupling between these potentials. Significant (P < 0.05, Bonferroni-corrected) cross-covariances were calculated between electrodes of lateral and median scalp regions. For both delta- and theta bandpassed potentials, covariance modeling indicated a shifting functional coupling between contralateral and ipsilateral frontal-central-parietal scalp regions and between these two regions and the median frontal-central scalp region from the preparation to the execution of the movement (P < 0.05). A maximum inward functional coupling of the contralateral with the ipsilateral frontal central-parietal scalp region was modeled during the preparation and initiation of the movement, and a maximum outward functional coupling during the movement execution. Furthermore, for theta-bandpassed potentials, rapidly oscillating inward and outward relationships were modeled between the contralateral frontal central-parietal scalp region and the median frontal-central scalp region across the preparation, initiation, and execution of the movement. We speculate that these cross-covariance relationships might reflect an oscillating dynamic functional coupling of primary sensorimotor and supplementary motor areas during the planning, starting, and performance of unilateral movement. The involvement of these cortical areas is supported by the observation that averaged spatially enhanced delta- and theta-bandpassed potentials were computed from the scalp regions where task-related electrical activation of primary sensorimotor areas and supplementary motor area was roughly represented. PMID- 9741748 TI - An expert system for EEG monitoring in the pediatric intensive care unit. AB - OBJECTIVES: was to design a warning system for the pediatric intensive care unit (PICU). The system should be able to make statements at regular intervals about the level of abnormality of the EEG. The warnings are aimed at alerting an expert that the EEG may be abnormal and needs to be examined. METHODS: A total of 188 EEG sections lasting 6 h each were obtained from 74 patients in the PICU. Features were extracted from these EEGs, and with the use of fuzzy logic and neural networks, we designed an expert system capable of imitating a trained EEGer in providing an overall judgment of abnormality about the EEG. The 188 sections were used in training and testing the system using the rotation method, thus separating training and testing data. RESULTS: The EEGer and the expert system classified the EEGs in 7 levels of abnormality. There was concordance between the two in 45% of cases. The expert system was within one abnormality level of the EEGer in 91% of cases and within two levels in 97%. CONCLUSIONS: We were therefore able to design a system capable of providing reliably an assessment of the level of abnormality of a 6 h section of EEG. This system was validated with a large data set, and could prove useful as a warning device during long-term ICU monitoring to alert a neurophysiologist that an EEG requires attention. PMID- 9741749 TI - Generalized background qEEG abnormalities in localized symptomatic epilepsy. AB - Spectral features of EEG background activity were studied in patients with localized symptomatic epilepsy (LSE), with origin in the frontal or temporal lobes. Z-values of high resolution spectra and measures of the parametric (xi alpha) model of the EEG were obtained for all 10/20 System leads and were compared with those obtained in a control group. Comparisons were performed between syndromic variants of LSE and between subgroups of patients with or without paroxysmal activity in their digital EEGs (dEEG). Marked reduction of the energy in the alpha range and a mild increase in the theta range were found in the patients, unrelated to the syndromic variant of the epilepsy. These deviations from normality were widespread on the scalp and were not related to antiepileptic medication. Non-parametric testing showed a positive correlation between the magnitude of the quantitative EEG abnormalities and the amount of paroxysmal activity in the dEEG. Slowing of the mean frequency of alpha components of the spectra, an actual decrease of power in the alpha range and an increase in the theta range explained the results. The most striking finding of this paper is that focal epileptogenesis may have a generalized impact in the frequency composition of EEG. PMID- 9741750 TI - Non-invasive epileptic focus localization using EEG-triggered functional MRI and electromagnetic tomography. AB - We present a new approach for non-invasive localization of focal epileptogenic discharges in patients considered for surgical treatment. EEG-triggered functional MR imaging (fMRI) and 3D EEG source localization were combined to map the primary electrical source with high spatial resolution. The method is illustrated by the case of a patient with medically intractable frontal lobe epilepsy. EEG obtained in the MRI system allowed triggering of the fMRI acquisition by the patient's habitual epileptogenic discharges. fMRI revealed multiple areas of signal enhancement. Three-dimensional EEG source localization identified the same active areas and provided evidence of onset in the left frontal lobe. Subsequent electrocorticography from subdural electrodes confirmed spike and seizure onset over this region. This approach, i.e. the combination of EEG-triggered fMRI and 3D EEG source analysis, represents a promising additional tool for presurgical epilepsy evaluation allowing precise non-invasive identification of the epileptic foci. PMID- 9741751 TI - Time-frequency analysis using the matching pursuit algorithm applied to seizures originating from the mesial temporal lobe. AB - OBJECTIVES: The ability to analyze patterns of recorded seizure activity is important in the localization and classification of seizures. Ictal evolution is typically a dynamic process with signals composed of multiple frequencies; this can limit or complicate methods of analysis. The recently-developed matching pursuit algorithm permits continuous time-frequency analyses, making it particularly appealing for application to these signals. The studies here represent the initial applications of this method to intracranial ictal recordings. METHODS: Mesial temporal onset partial seizures were recorded from 9 patients. The data were analyzed by the matching pursuit algorithm were continuous digitized single channel recordings from the depth electrode contact nearest the region of seizure onset. Tine frequency energy distributions were plotted for each seizure and correlated with the intracranial EEG recordings. RESULTS: Periods of seizure initiation, transitional rhythmic bursting activity, organized rhythmic bursting activity and intermittent bursting activity were identified. During periods of organized rhythmic bursting activity, all mesial temporal onset seizures analyzed had a maximum predominant frequency of 5.3-8.4 Hz with a monotonic decline in frequency over a period of less than 60 s. The matching pursuit method allowed for time-frequency decomposition of entire seizures. CONCLUSIONS: The matching pursuit method is a valuable tool for time frequency analyses of dynamic seizure activity. It is well suited for application to the non-stationary activity that typically characterizes seizure evolution. Time-frequency patterns of seizures originating from different brain regions can be compared using the matching pursuit method. PMID- 9741752 TI - Volume conduction effects in EEG and MEG. AB - Volume conductor models that are commonly used to describe the EEG and MEG neglect holes in the skull, lesions, the ventricles, and anisotropic conductivity of the skull. To determine the influence of these features, simulations were carried out using the finite element method. The simulations showed that a hole in the skull will have a large effect on the EEG, and as one of the consequences localisation errors up to 15 mm may occur. The effect on the MEG is negligible. The presence of a lesion may cause the shape and magnitude of the EEG and MEG to change. Hence, a lesion has to be taken into account, if the active neurones are close to it. Moreover, a localisation procedure may fail if the lesion is not included in the volume conductor model. Inclusion of the ventricles in the volume conductor model is necessary only if sources are in their vicinity or if their sizes are unusually large. Anisotropic conductivity of the skull has a smearing effect on the EEG but does not influence the MEG. PMID- 9741753 TI - Event-related electric microstates of the brain differ between words with visual and abstract meaning. AB - The present study shows that different neural activity during mental imagery and abstract mentation can be assigned to well-defined steps of the brain's information-processing. During randomized visual presentation of single, imagery type and abstract-type words, 27 channel event-related potential (ERP) field maps were obtained from 25 subjects (sequence-divided into a first and second group for statistics). The brain field map series showed a sequence of typical map configurations that were quasi-stable for brief time periods (microstates). The microstates were concatenated by rapid map changes. As different map configurations must result from different spatial patterns of neural activity, each microstate represents different active neural networks. Accordingly, microstates are assumed to correspond to discrete steps of information processing. Comparing microstate topographies (using centroids) between imagery- and abstract-type words, significantly different microstates were found in both subject groups at 286-354 ms where imagery-type words were more right-lateralized than abstract-type words, and at 550-606 ms and 606-666 ms where anterior posterior differences occurred. We conclude that language-processing consists of several, well-defined steps and that the brain-states incorporating those steps are altered by the stimuli's capacities to generate mental imagery or abstract mentation in a state-dependent manner. PMID- 9741754 TI - Digital archival and exchange of events in a simple format for polygraphic recordings with application in event related potential studies. AB - This paper describes a simple method of event encoding as an extension to a previously defined standard format, the European Data Format (EDF). The specification ensures full backward compatibility with the existing definition. By using this extension, the format can be used to store both continuous recordings and selected epochs of recordings. The encoding is performed in a channel of event-codes or in a pseudo-channel for annotations. Standardisation of event encoding is discussed. Decoding of events or annotations from the extended format is implemented at the application level. Existing programs that do not support the new encoding scheme still operate correctly and can simply ignore the new channels in processing 'extended' data files. The event encoding is also compatible with EDF's capability to encode channels of different sampling frequency. PMID- 9741755 TI - Non-invasive piezoelectric transducer for recording of respiration at the level of diaphragm. AB - In this study, piezoelectric polyvinylidenefluoride (PVDF) wire was used in construction of a transducer for recording of respiration. The recordings have been performed in 202 subjects, mainly during normal routine electroencephalography (EEG) examinations. Comparison of the PVDF wire transducer with the piezoelectric Siemens Movement Sensor 230 was performed. The recordings of respiration showed highly similar curves with both methods. The PVDF wire transducer seems to record the breathing movements at the level of the diaphragm with great reliability in EEG-polygraphy and it is inexpensive, easy to use and to clean. PMID- 9741756 TI - A rapid method for determining standard 10/10 electrode positions for high resolution EEG studies. AB - This report describes the basic principle and examines the comparative accuracy of a novel method for locating 3-D coordinates of electrode positions on the head. The method involves calculation of the 3-D coordinates for any array of 10/10 electrode positions from 14 straight-line distances between 11 10/10 electrodes. In 11 subjects the 3-D coordinates of 64 scalp electrodes embedded in an electrode cap were identified with the novel method, and also with a standard commercial magnetic field digitizer. The outcomes from the two methods were compared with directly measured coordinates of all 64 positions (cf. De Munck, J.C., Vijn, P.C.M. and Spekreijse, H. A practical method for determining electrode positions on the head. Electroenceph. clin. Neurophysiol., 1991, 89: 85 87). Coordinates in 3 dimensions obtained using the new method were significantly closer to the directly measured values than those from the magnetic field digitizer. The new method was also quicker and requires less specialized instrumentation than the magnetic field digitization method. The novel method appears to be a valid and convenient tool for use with EEG analysis techniques that require specific information about 10/10 electrode positions. PMID- 9741757 TI - The organization of the cortical motor system: new concepts. AB - A series of recent anatomical and functional data has radically changed our view on the organization of the motor cortex in primates. In the present article we present this view and discuss its fundamental principles. The basic principles are the following: (a) the motor cortex, defined as the agranular frontal cortex, is formed by a mosaic of separate areas, each of which contains an independent body movement representation, (b) each motor area plays a specific role in motor control, based on the specificity of its cortical afferents and descending projections, (c) in analogy to the motor cortex, the posterior parietal cortex is formed by a multiplicity of areas, each of which is involved in the analysis of particular aspects of sensory information. There are no such things as multipurpose areas for space or body schema and (d) the parieto-frontal connections form a series of segregated anatomical circuits devoted to specific sensorimotor transformations. These circuits transform sensory information into action. They represent the basic functional units of the motor system. Although these conclusions mostly derive from monkey experiments, anatomical and brain imaging evidence suggest that the organization of human motor cortex is based on the same principles. Possible homologies between the motor cortices of humans and non-human primates are discussed. PMID- 9741758 TI - Localisation of epileptic foci with electric, magnetic and combined electromagnetic models. AB - We compare the localisation of epileptic foci by means of (1) EEG, (2) magnetoencephalography (MEG) and (3) combined EEG/MEG data in a group of patients suffering from pharmaco-resistant focal epilepsy. Individual epileptic events were localised by means of a moving dipole model in a 4-shell spherical head approximation. A patient's epileptic activity was summarised by calculating the spatial density distribution (DD) of all localised events, and the centre of gravity of DD was considered the most likely locus of seizure generation. To verify these loci a subgroup of 6 patients was selected, in which seizures could be related to a clearly identifiable lesion in MRI. On average, the combined EEG/MEG approach resulted in the smallest error (1.8 cm distance between calculated locus and the nearest lesion border); using only MEG yielded the largest error (2.4 cm), while EEG resulted in an intermediate value (2.2 cm). In the individual patients, EEG/MEG would also rank intermediate, but never worst. In summary, combining EEG/MEG appears to be a more robust approach to localisation than using only EEG or only MEG. Finally, we also report on the use of the barbiturate methohexital as a safe method of increasing the number of spike events during an EEG/MEG recording session. PMID- 9741759 TI - Dipole source localization by means of maximum likelihood estimation I. Theory and simulations. AB - By analyzing simulated neuromagnetic recordings with a 37 channel magnetometer system, it was shown that the accuracy of a dipole source localization is considerably improved if the standard least-squares fit procedure is replaced by a maximum likelihood estimation accounting for the covariances of the noise in the measurement channels. Spatially correlated noise was generated by random dipoles homogeneously distributed in a sphere representing the brain. The study suggests that a maximum likelihood estimation reduces the standard deviations of the estimated dipole parameters by roughly a factor of two. PMID- 9741760 TI - Dipole source localization by means of maximum likelihood estimation. II. Experimental evaluation. AB - A dipole source analysis of repeated measurements of the auditory evoked field (AEF) elicited by 60 dB SL tonebursts confirmed the previous theoretical prediction that a maximum likelihood estimation technique reduces localization errors resulting from noise in the measured data by about a factor of two as compared to the common least-squares fit procedure. To achieve a comparable improvement by averaging independent epochs (Gaussian noise assumed), the number of averages would have to be increased by a factor of 4. Conversely, the measurement time required to achieve a given localization accuracy could be reduced by that factor. PMID- 9741761 TI - In the footsteps of Beck: the desynchronization of the electroencephalogram. PMID- 9741762 TI - Improved realistic Laplacian estimate of highly-sampled EEG potentials by regularization techniques. AB - In this study we investigated the effects of lambda correction, generalized cross validation (GCV), and Tikhonov regularization techniques on the realistic Laplacian (RL) estimate of highly-sampled (128 channels) simulated and actual EEG potential distributions. The simulated EEG potential distributions were mathematically generated over a 3-shell spherical head model (analytic potential distributions). Noise was added to the analytic potential distributions to mimic EEG noise. The magnitude of the noise was 20, 40 and 80% that of the analytic potential distributions. Performance of the regularization techniques was evaluated by computing the root mean square error (RMSE) between regularized RL estimates and analytic surface Laplacian solutions. The actual EEG data were human movement-related and short-latency somatosensory-evoked potentials. The RL of these potentials was estimated over a realistically-shaped, magnetic resonance constructed model of the subject's scalp surface. The RL estimate of the simulated potential distributions was improved with all the regularization techniques. However, the lambda correction and Tikhonov regularization techniques provided more precise Laplacian solutions than the GCV computation (P < 0.05); they also improved better than the GCV computation the spatial detail of the movement-related and short-latency somatosensory-evoked potential distributions. For both simulated and actual EEG potential distributions the Tikhonov and lambda correction techniques provided nearly equal Laplacian solutions, but the former offered the advantage that no preliminary simulation was required to regularize the RL estimate of the actual EEG data. PMID- 9741763 TI - Segmentation and classification of EEG during epileptic seizures. AB - We present a method for the automatic comparison of epileptic seizures in EEG, allowing the grouping of seizures having similar overall patterns. Each channel of the EEG is first broken down into segments having relatively stationary characteristics. Features are then calculated for each segment and all segments of all channels of the seizures of one patient are grouped into clusters of similar morphology. This clustering allows labeling of every EEG segment. Methods derived from string matching procedures are then used to obtain an overall edit distance between two seizures, a distance that represents how the two seizures, taken in their entirety and including the channels not actually involved in the discharge, resemble each other. Examples from 5 patients, 3 with intracerebral electrodes and two with scalp electrodes, illustrate the ability of the method to group seizures of similar morphology. PMID- 9741764 TI - Electrocorticography and outcome in frontal lobe epilepsy. AB - The prognostic significance of epileptiform activity (EA) recorded at electrocorticography (ECOG) was examined in a group of 60 consecutive non-tumoral patients with intractable frontal lobe epilepsy (FLE). Pre-excision EA was documented as absent, focal (one gyrus), regional (two gyri), lobar (3 gyri) or multilobar (frontal + temporal gyri). Post-excision EA was documented as absent, restricted to the resection border, or recorded distant to the resection border, and was quantitated by spike frequency. Pre-excision EA from < or = 2 gyri and absence of post-resection EA correlated with Class I or II (Engel classification) outcome while pre-excision EA from > or = 3 gyri and persistent post-resection EA, especially distant to the resection border, correlated with Class III or IV outcome (P < 0.001). A significant correlation between poorer outcomes and increased abundance of distant post-resection EA was observed (P < 0.001). EA restricted to the resection border was not significantly correlated with outcome. Presence of a circumscribed lesion correlated with Class I outcome (P < 0.01) and absence of pathological abnormality correlated with Class IV outcome (P < 0.05). Neither side nor extent of surgical excision correlated with outcome. EA recorded at ECOG is of prognostic significance in FLE. A lobar or multilobar distribution of pre-excision EA and persistent post-excision EA distant to the resection border, especially when abundant, are highly unfavorable prognostic indicators. In contrast, a restricted distribution of pre-excision EA and absence of post resection EA both herald a favorable outcome. PMID- 9741765 TI - Interictal epileptiform activity in elderly patients with epilepsy. AB - OBJECTIVE: To examine the frequency of interictal epileptiform activity (IEA) in elderly patients with epilepsy. DESIGN AND METHODS: From a consecutive 13,905 EEGs recorded over 5 years at a university hospital EEG laboratory, 558 studies were performed on outpatients aged 60 years or more. Medical record review identified 125 patients in whom a confident clinical diagnosis of epilepsy was made by a board-certified neurologist. The EEG findings in these patients were reviewed. The effects of various variables on the likelihood of detecting IEA were calculated using Fisher's test and chi-square analysis. RESULTS: IEA were present on the first EEG in 35% of 55 patients (mean age 65 years) with pre existing epilepsy, and 26% of 70 patients (mean age 70 years) with seizure onset after 60 years. There were no significant differences in the frequency of IEA in patients with late onset epilepsy in the 7th or in the 8th decades of life. Most IEA were focal. Activation procedures added little additional information. Patients with more than one seizure per month were significantly more likely to have IEA (P = 0.016). There were no major differences in likelihood of IEA detection depending on the underlying cause of the seizures. CONCLUSIONS: The frequency of IEA in elderly patients with epilepsy is substantially lower than that reported in epileptic populations as a whole. This low rate of IEA in routine EEG studies must be recognized when considering the diagnosis of an epileptic syndrome for episodic events happening in the elderly. PMID- 9741767 TI - Quantitative electroencephalography in amyotrophic lateral sclerosis. AB - OBJECTIVES: The authors evaluated quantitative EEG (QEEG) in patients with amyotrophic lateral sclerosis (ALS), in order to see if the confined cortical degeneration found in anatomical and functional examinations of central (rolandic) regions could give rise to abnormalities of cortical electrical activity. MATERIAL AND METHODS: Eighteen patients with ALS were compared with 14 control subjects comparable for age and sex. The QEEG analysis was blind to the clinical data. Frequency analysis of 30 EEG epochs, each of 2.6 s duration, was performed. RESULTS: QEEG in ALS patients showed a significant well-localized decrease of alpha activity only in the central regions, while theta and delta relative power did not show any difference from controls. CONCLUSION: We suggest that QEEG is able to show evidence of subtle EEG changes probably due to loss of cells in the somatomotor cortex selectively affecting the generator sites. PMID- 9741766 TI - Does pattern electroretinogram spatial tuning alteration in Parkinson's disease depend on motor disturbances or retinal dopaminergic loss? AB - Systemic decrease of dopaminergic cells, such as in Parkinson's disease may produce visual alterations in humans. In order to show possible pattern electroretinogram (PERG) spatial tuning function (STF) alterations due to impaired dopaminergic transmission in humans, we studied a group of Parkinson's disease patients before and during treatment with the dopamine precursor, levodopa, and compared their performances with those of an age-matched control group. Moreover, in order to exclude the possible involvement of motor disabilities to produce PERG alterations, we also investigated PERG responses in post-traumatic parkinsonian patients who exhibited motor abnormalities as a consequence of focal lesions of basal ganglia, in the absence of systemic dopaminergic degeneration. Our results showed a clear decrease of PERG responses in Parkinson's disease patients particularly at medium spatial frequency range (2.7-4.0 cycles/degree) with a substantial preservation of responses at low frequencies. Levodopa therapy reversed these alterations in Parkinson's disease patients, resulting in the recovery of a normal tuning function shape. In contrast to Parkinson's disease, the tuning function appeared to be preserved in post-traumatic parkinsonian patients. Our results clearly establish a relationship between retinal alteration in PD patients and dopaminergic retinal function. PMID- 9741768 TI - Animal electricity from Bologna to Boston. AB - This is an appreciation of 3 scientists who made historic contributions toward understanding bio-electrical activity. The discoveries of Galvani and Volta, who were contemporaries two hundred years ago, continue as basic supports in advancing the strength and health of all mankind. They, nevertheless, had political and scientific disagreements that still linger. The third scientist was our contemporary, Alexander Forbes who, throughout most of the 20th century, continued to increase our understanding of electrical activity in the nervous system. PMID- 9741769 TI - The fundamental neural mechanisms of electroencephalography. AB - We are at an interesting time in the evolution of the EEG. Studies are opening the door to understanding the intrinsic neuronal properties and network operations responsible for the generation of EEG oscillations. I will review some of our knowledge regarding the physiology of the normal and abnormal EEG. Both epileptic and non-epileptic activity will be discussed. Less is known about the latter, because of difficulties in developing appropriate models. The major dichotomy for both types of EEG phenomenon will be focal and generalized (or widespread. Certain distinctive abnormal EEG patterns including burst suppression, periodic phenomena and intermittent rhythmic delta will also be addressed. PMID- 9741770 TI - Common applications of electrophysiology (EEG) in the past and today: the technologist's view. AB - The field of electroencephalography (EEG) has witnessed a dramatic development during the last decade. The electroencephalogram that had been principally used as a 'post-hoc' diagnostic procedure is now fully used as an 'on-line' monitor of neural function with its excellent temporal resolution. Neurophysiological monitoring in the operating room, neurological intensive care unit (ICU) and during endovascular procedures allows early identification of impending neurological deficits before irreversible neurological impairment. Long-term monitoring of scalp and synchronized video-EEG recordings and invasive (depth and subdural) electrode studies are well-established tools in the localization of the epileptogenic region for identification of potential candidates for surgery. The advent of digital EEG with digital storage and the ability to manipulate data with digital reformatting, filter and sensitivity changes has allowed us to maximize the information and reduce artifacts. These changes have revolutionized the way in which EEG is performed and interpreted. PMID- 9741771 TI - The advantages of digital over analog recording techniques. AB - In the past few years digital technology has brought EEG and evoked potentials into emergency rooms, intensive care units and operating rooms with a variety of automatic data trending. Networking of these systems makes access to clinical neurophysiologists nearly immediate. Digital EEG has made montage reformatting and quantitation of parameters readily available. Increased spatial and temporal resolution is available with routine EEG, and combined topographic and frequency mapping of a given potential, spike or seizure focus is possible. In the future, these and other features such as dipole mapping and cognitive EP analysis will be available on a routine basis. PMID- 9741772 TI - Computer-based electroencephalography: technical basics, basis for new applications, and potential pitfalls. AB - EEG has been recorded on paper-based analog systems for over 50 years. In the past 5 years, computer-based digital systems have become more widely used. Digital systems eliminate some artifacts that plagued analog recordings but introduce subtle new problems including aliasing and dynamic range. Digital systems allow reformatting of the same EEG segment using different gain, filter and montage settings. The digital signal allows for measurement and computations on the EEG, leading to applications such as power spectrum, topographic mapping, and spike or seizure detection. PMID- 9741773 TI - Trends in EEG source localization. AB - The concepts underlying the quantitative localization of the sources of the EEG inside the brain are reviewed along with the current and emerging approaches to the problem. The concepts mentioned include monopolar and dipolar source models and head models ranging from the spherical to the more realistic based on boundary and finite elements. The forward and inverse problems in electroencephalography are discussed, including the non-uniqueness of the inverse problem. The approaches to the solution of the inverse problem described include single and multiple time-slice localization, equivalent dipole localization and the weighted minimum norm. The multiple time-slice localization approach is highlighted as probably the best available at this time and is discussed in terms of the spatiotemporal model of the EEG. The effect of noise corruption, artifacts and the number of recording electrodes on the accuracy of source localization is also mentioned. It is suggested that the main appeal of the minimum norm is that it does not assume a model for the sources and provides an estimate of the current density everywhere in the three dimensional volume of the head. PMID- 9741774 TI - Potential fields, EEG maps, and cortical spike generators. AB - From analysis of the interictal discharge, one can deduce the location of the underlying generator or source. Despite the geometric complications of cortical convolutions and major fissures, clinical neurophysiologists fearlessly make statements about localization of the source. In fact, this is usually nothing more than a qualitative dipole localization, which is seldom discussed with the concern of non-unique mathematical solutions. However, further information can be gleaned from the spike topography if one applies some simple constraints or assumptions (discrete point source, columnar organization). An explanation of the tangential and radial fields of centrotemporal spikes is offered, together with an attempt to rationalize why the tangential topography is associated with a more benign clinical picture than the radial topography. It is suggested that there is differential epileptogenicity between gyral and fissural cortex. Extension of this is made to multiple subpial transection. PMID- 9741775 TI - Fundamentals of evoked potentials and common clinical applications today. AB - Visual, auditory and somatosensory evoked potentials are commonly used in neurology today to confirm and localize sensory abnormalities, to identify silent lesions and to monitor changes. Methods have become standardized. Normal limits are now well described. Published reports have described well how these evoked potentials are different in various types of neurologic disorder. Intensive care unit applications and surgical monitoring have also developed appropriate medical uses of these tests. Evoked potentials have become useful as they are relatively objective, reproducible, very sensitive to impairment and relatively easy to use in many clinical settings. PMID- 9741776 TI - Evaluation and prognostication in coma. AB - Electroencephalography (EEG) and evoked potential (EP) studies are neurophysiologic techniques which provide information on physiological state and response to therapy, and may aid diagnosis and prognosis. Serial studies or continuous monitoring may enable changes to be detected prior to irreversible deterioration in the patient's condition. Current computer technology allows simultaneous display and correlation of electrophysiologic parameters, cardiovascular state and ICP. Continuous EEG monitoring in the ICU has been shown to have a decisive or contributing impact on medical decision making in more than three-quarters of patients. In addition, continuous EEG monitoring has revealed previously unsuspected non-convulsive seizures in two-thirds of patients. Somatosensory and auditory EPs can provide useful prognostic information in coma patients, however, these tests are etiologically non-specific and must be carefully integrated into the clinical situation. Motor EPs offer a potentially useful tool for evaluating motor system abnormalities in the ICU. Thus, neurophysiologic tests are established monitoring tools in the neurological intensive care unit. PMID- 9741777 TI - Generators of the late cognitive potentials in auditory and visual oddball tasks. AB - Recordings directly within the brain can establish local evoked potential generation without the ambiguities always associated with extracranial electromagnetic measures. Depth recordings have found that sensory stimuli activate primary cortex and then material-specific encoders. Sensory-specific areas remain active for long periods, but by about 200 ms are joined by activation in widespread brain systems. One system is related to the orientation of attention. It is centered in paralimbic and attentional frontoparietocingular cortex, and associated with the P3a. A second system associated with P3b envelopes cognitive contextual integration. It engages the ventral temporofrontal event-encoding cortices (inferotemporal, perirhinal, and ventrolateral prefrontal), association cortices (superior temporal sulcal and posterior parietal), and the hippocampus. Thus, even in simple tasks, activation is widespread but concentrated in particular multilobar systems. With this information, the late cognitive potentials can be used to monitor the probable location, timing and intensity of brain activation during cognitive tasks. PMID- 9741779 TI - Timeline of the history of EEG and associated fields. PMID- 9741778 TI - The future of electroencephalography in assessing neurocognitive functioning. AB - High temporal resolution is necessary to resolve the rapidly changing patterns of brain activity underlying mental function. Additionally, simple, non-intrusive equipment is needed to routinely measure such functions in doctors' offices, at home and work and in other naturalistic contexts as people perform normal everyday activities. When compared with all other modalities for measuring higher brain functions, EEG is unique in that it has both these attributes. Two factors are limiting the further development and application of EEG for measuring cognitive functioning: a technical one that is easy to overcome and a sociological one that is more problematic. The technical limitation is that traditional EEG technology and practice provides insufficient spatial detail to identify relationships between brain electrical events and structures and functions visualized by magnetic resonance imaging (MRI) or other modalities. Recent advances overcome this problem by recording EEGs from more electrodes, by registering EEG data with anatomical information from each subject's MRI, by correcting the distortion caused by volume conduction of EEG signals through the skull and scalp, and by computing hypotheses about the sources of signals recorded at the scalp. The sociological limitation is that clinical EEGs are mostly performed by neurologists with no particular special interest in cognitive brain function, while cognitive research using EEG is largely done by psychology professors and their graduate students with no clinical ambitions. The diminishing clinical role of traditional EEGs in localizing lesions in the brain, and the obvious and insistent medical need for inexpensive and accessible tests of cognitive brain functioning may serve to soon dissipate this sociological obstruction. This will lead to a golden age of EEG in which Hans Berger's vision of the EEG as a window on the mind will be realized. Rather than slowly fading into obsolescence, EEG will retain its role as the primary means of measuring higher brain function when the purpose is not 3D localization per se, and will serve as an invaluable complement to functional MRI in those instances when both high temporal and high spatial resolution are required. PMID- 9741780 TI - Tamoxifen--the treatment of choice. Why look for alternatives? AB - Tamoxifen is currently established as the endocrine treatment of choice in breast cancer. In advanced breast cancer, response rates of up to 60% in women with oestrogen receptor (ER)-positive tumours have been reported. In early breast cancer, tamoxifen can produce significant benefits, both statistically and clinically, in terms of reduction in relative risk of relapse or death in all patient subgroups (i.e. ER status, aged < or > 50 years) except premenopausal women with ER-negative tumours. The major benefit, however, is seen in women over 50 years old with ER-positive tumours. The results of randomized trials suggest that the optimum duration of tamoxifen therapy is at least 5 years. Two large pragmatic trials (aTTom and ATLAS) are under way to determine whether additional benefit can be gained from continuing tamoxifen treatment beyond 5 years. Recent data also suggest possible synergism between tamoxifen and chemotherapy in the treatment of early breast cancer in post-menopausal women. Other benefits of tamoxifen treatment include reduction in the risk of developing contralateral breast cancer. Included among the non-breast cancer benefits of tamoxifen are reduced risk of cardiovascular disease and protection against bone loss in post menopausal women. These benefits must be weighed against the possible increased incidence of endometrial cancer. Notwithstanding its undoubted success, there is a need for agents to improve upon tamoxifen. Newer agents, such as the luteinizing hormone-releasing hormone analogue goserelin and the new-generation aromatase inhibitors, such as anastrozole, will add new life to the search for an improved endocrine therapy for early breast cancer. PMID- 9741781 TI - Luteinizing hormone-releasing hormone analogues--the rationale for adjuvant use in premenopausal women with early breast cancer. AB - Current standard adjuvant therapies for early breast cancer include tamoxifen and chemotherapy, depending on the disease prognosis and menopausal status. Luteinizing hormone-releasing hormone (LHRH) analogues offer a different approach to the management of early breast cancer in pre- and perimenopausal women. The most widely studied LHRH analogue is goserelin. It acts on the hypothalamic pituitary axis to suppress ovarian function, decreasing luteinizing hormone and oestradiol levels to post-menopausal values. Pooled data from 228 premenopausal and perimenopausal patients with advanced breast cancer enrolled in 29 studies worldwide demonstrated an objective response rate for goserelin, 3.6 mg, of 36.4%, with a median duration of response of 44 weeks. These results fall well within the ranges of reported response rates for ovarian ablation and for tamoxifen in similar patient populations. By virtue of its mode of action, goserelin does not stimulate the ovaries and is unlikely to have detrimental effects on the endometrium. In addition, given that goserelin has no oestrogen agonist-like effects, unlike tamoxifen, there is no potential for tumour stimulation in those patients becoming resistant to treatment. Goserelin is generally well tolerated, and the main side-effects are related to ovarian suppression, which is potentially reversible. Preliminary results in premenopausal women with early breast cancer indicate that endocrine treatment with goserelin plus tamoxifen may be as effective as standard combination chemotherapy (cyclophosphamide-methotrexate-5-fluorouracil), but has significantly less acute toxicity. A number of large, randomized trials are now in progress to assess the potential role of goserelin as adjuvant therapy for early breast cancer. PMID- 9741782 TI - Luteinizing hormone-releasing hormone analogues in early breast cancer: updated status of ongoing clinical trials. AB - In the year 2000, the ongoing meta-analysis of the Early Breast Cancer Trialists' Collaborative Group will be updated to include additional data from over 4000 patients treated with luteinizing hormone-releasing hormone analogues, principally goserelin. Four major international trials are currently in progress to evaluate the safety and efficacy of goserelin in comparison with the current standard treatments in early breast cancer, which are chemotherapy or tamoxifen. This paper provides an outline of the protocols and main objectives of the Zoladex Early Breast Cancer Research Association (ZEBRA) trial (goserelin versus cyclophosphamide-methotrexate-5-fluorouracil [CMF]), the Cancer Research Campaign (CRC) trial (goserelin versus tamoxifen versus the combination of goserelin and tamoxifen versus no further treatment), the International Breast Cancer Study Group (IBCSG) VIII trial (goserelin versus CMF versus CMF followed by goserelin) and the Eastern Cooperative Oncology Group (ECOG)/South Western Oncology Group (SWOG) trial (cyclophosphamide-doxorubicin-5-fluorouracil [CAF] versus CAF followed by goserelin versus CAF followed by goserelin plus tamoxifen). Preliminary results are expected from the CRC trial in 1998 and from the ZEBRA and ECOG/SWOG trials in 1999. Results from the wide range of comparator regimens, treatment durations and patient subgroups investigated in these trials will greatly increase the clinical database and should help to define the optimum role for goserelin in the treatment of early breast cancer in premenopausal women. PMID- 9741783 TI - Aromatase inhibitors and their future role in post-menopausal women with early breast cancer. AB - Anastrozole is the first aromatase inhibitor to show a significant survival advantage over megestrol acetate in post-menopausal women with advanced breast cancer. The rationale for extending the use of aromatase inhibitors to the treatment of early breast cancer is based on the efficacy observed in the advanced setting, combined with good tolerability and a convenient dosing regimen. Furthermore, oestrogen deprivation by ovarian ablation (similar to oestrogen antagonism with tamoxifen) is already established as an effective adjuvant treatment in premenopausal women with modality breast cancer. Anastrozole produces a profound suppression of plasma oestrogen levels which is greater than that obtained with earlier aromatase inhibitors (formestane, aminoglutethimide) or megestrol acetate. This could account for the differences in clinical efficacy seen between anastrozole and megestrol acetate. In terms of benefits over other endocrine agents, anastrozole causes significantly less weight gain than megestrol acetate; it does not have the partial agonist activity of tamoxifen, and is unlikely to lead to tumour stimulation in patients resistant to tamoxifen or to exert proliferative effects on the endometrium. The lack of oestrogen agonist activity, however, may possibly have detrimental effects on bone mineral density and blood lipid profile. Current clinical trials are investigating the efficacy and safety of anastrozole in the early breast cancer setting. The results of these trials will help to determine whether anastrozole has any benefits over tamoxifen, the current treatment of choice in post menopausal women with early breast cancer. PMID- 9741784 TI - The place of chemotherapy in the treatment of early breast cancer. AB - The choice of systemic treatment for breast cancer depends on the tumour characteristics and stage of disease, and the patient's age, general state of health, menopausal status and oestrogen receptor (ER) status. Traditionally, endocrine therapy has been reserved for post-menopausal women, combination chemotherapy being more commonly used in premenopausal women. Chemotherapy remains the only option for patients with ER-negative breast cancer. The 1992 EBCTCG overview showed that, overall, polychemotherapy as adjuvant treatment for early breast cancer produced significant reductions in annual odds of recurrence and mortality, with a statistically significant trend towards greater benefits in patients aged under 50 years. Several trials have shown combination chemotherapy with cyclophosphamide, methotrexate and 5-fluorouracil (CMF) to be more effective than single-agent chemotherapy in premenopausal women with node-positive tumours. However, although CMF chemotherapy seems to be effective irrespective of menopausal status, this benefit appears greatest in premenopausal women. The addition of anthracyclines to combination chemotherapy regimens has extended disease-free and overall survival rates in both premenopausal and post-menopausal women, including those with ER-positive tumours. The use of high-dose chemotherapy with stem cell support in early breast cancer is unjustified outside the clinical trial setting--current data indicate that such treatment may result in increased morbidity without a reduction in disease recurrence. Tamoxifen is effective in ER-positive disease; however, as yet few large comparative trials have compared endocrine treatment with chemotherapy in early breast cancer. Combination chemoendocrine therapy may provide a greater benefit than tamoxifen alone in early breast cancer, but this requires further study. PMID- 9741785 TI - Evident trans-synaptic degeneration of motor neurons after stroke: a study of neuromuscular jitter by axonal microstimulation. AB - Neuronal degradation accompanied by axonal degeneration has been known to occur in lower motor neurons following a stroke. In the present study, the functional integrity of neuromuscular transmission was assessed, utilizing a sensitive electrodiagnostic method consisting of stimulated single-fiber electromyography (SFEMG), along with axonal microstimulation, in paralytic muscles of stroke patients. Neuromuscular jitter was measured in the hemiplegic side extensor digitorum communis (EDC) as well as in anterior tibial (AT) muscles for 28 stroke patients and also for 13 age-matched controls. The disease duration, i.e. from the onset of stroke until the stimulated SFEMG examination, extended from 2 months to 8 years. Mean jitters obtained in EDC and AT muscles of stroke patients were found to be significantly greater than those in normal controls. Mean jitters obtained in severely weak muscles of stroke patients were greater than those in moderately weak muscles. Positive correlations were noted between the increased jitter and the disease duration from the onset of stroke until the time of the stimulated SFEMG test. These findings demonstrate a dysfunction of neuromuscular transmission in the paralytic muscles of stroke patients and suggests that trans-synaptic degeneration of motor neurons may occur in stroke. Furthermore, the neuronal degradation in stroke was positively correlated with the course duration of the disease. PMID- 9741786 TI - Force regulation is deficient in patients with parietal lesions: a system analytic approach. AB - By means of a quantitative system-analytic investigation strategy, the postural motor control of the fingers was evaluated, to characterise the possible deficit of force regulation in patients with parietal lesions. In spite of a normal response to short torque pulses, the parietal-lesion patients had difficulties in returning to the preload level after the application of an additional step torque load to fingers II-IV of their left or right hands. The control offset (measured 500 ms after step torque application) was significantly larger in the patient group. This deficit in the investigated patients with parietal lesions to compensate for step torque loads was not due to a paresis, but rather resulted from a disturbance in the generation of a sufficient counterforce against the applied step torque within an adequate time window and motor pattern. This distinct force-regulation deficit was found in patients with left- and right sided parietal lesions. PMID- 9741787 TI - Stumbling reactions in man: influence of corticospinal input. AB - The aim of this study was to evaluate the degree of contribution of supraspinal input to the generation of the compensatory leg muscle activation following stance perturbation. Therefore, evoked motor response (EMR) input-output relations of two different motor tasks were compared at 3 distinct periods: (1) the basic period of muscular activity during standing, i.e. when no additional cortical or spinal activity due to the different tasks is to be expected, (2) the pre-movement period with low background activity, when different spinal and cortical inputs to the motoneuronal pool can be assumed and (3) the period of plateau EMG activity of compensatory and voluntary motor task. Transcranial magnetic stimulation (TMS) just below the motor threshold was applied randomly at 19 different time-intervals before and during the onset of stance perturbation and for comparison during an equivalent voluntary foot-dorsiflexion task. Recordings of electromyographic (EMG) activity from the tibialis anterior (TA) and corresponding ankle-joint movements were made from both legs. Forward directed displacements were induced by randomly-timed ramp impulses of constant acceleration upon a moveable platform. For comparison, leg muscle EMG was recorded during isometric foot dorsiflexion during stance while leaning back against a support. The stance perturbations were followed by a compensatory response (CR) in the TA with a mean onset time of 81 ms. During the basic period of muscular activity and the period of plateau EMG activity there was no significant difference of the input-output relation between stance perturbation and the voluntary motor task. However, in the voluntary task compared with the CR, there was significantly greater input-output relation (facilitation) of the EMR in the TA following TMS, which may be related to an increased cortical influence. In contrast to this result of the CR following stance perturbation, a facilitation of the EMR was described for hand muscles under corresponding conditions of automatic compensation for muscle stretch, suggesting a transcortical reflex loop. This difference in the results from upper and lower extremity muscles favors the assumption of a predominantly spinal generation of the TA-CR following stance perturbation. PMID- 9741788 TI - Cortical silent period evoked by transcranial magnetic stimulation in ischemic stroke. AB - OBJECTIVES: Transcranial magnetic stimulation (TMS) of the motor cortex produces motor evoked potentials (MEPs). Besides this excitatory response, TMS has inhibitory effects. When TMS is performed during voluntary muscle contraction, the MEP is followed by a pause in electromyographic activity (cortical silent period, SP). The aim of this study was to evaluate the clinical usefulness of the SP. METHODS: We studied SP changes in 50 patients with acute hemispheric brain infarction. A stimulator with a round coil and a fixed intensity of 90% of maximum was used to evoke MEPs. RESULTS: SP was elicited on the affected side in 29 of the 50 patients. The mean SP duration was markedly longer on the affected side in the patient group. There were no significant differences between left and right sides in the means of the MEP amplitude ratio (amplitude related to corresponding amplitudes to peripheral electric stimulation) and MEP latencies in the patient group. Prolonged SP was found in 25 of the 29 patients (86%) whereas only 4 (14%) had abnormalities in MEP latency or amplitude ratio. The mean SP duration was significantly prolonged also in a subgroup of 14 patients with normal hand function. CONCLUSIONS: The SP measurement is an easily performed and sensitive method to detect even subclinical disturbances in motor system function in ischemic stroke. PMID- 9741789 TI - Interhemispheric inhibition in patients with multiple sclerosis. AB - OBJECTIVES: A single focal magnetic stimulus applied to the motor cortex of normal subjects can suppress ongoing voluntary electromyographic activity in ipsilateral small hand muscles. This inhibition is mediated from one motor cortex to the contralateral side via a transcallosal pathway. METHODS: We have investigated transcallosal inhibition in 24 patients with definite multiple sclerosis (MS) and in 24 healthy volunteers. A focal magnetic stimulus was applied to the hand area of the motor cortex and the onset latency of the inhibition of the ongoing EMG activity of the ipsilateral first dorsal interosseus muscle was evaluated. Cortico-motor conduction time to the same muscle was revealed, using a magnetic stimulus over the contralateral motor cortex. The difference between these values was calculated as transcallosal conduction time. Cerebral magnetic resonance imaging (MRI) scans including sagittal T2-weighted images were performed in 18 patients. RESULTS: The depth of inhibition (maximal inhibition as percentage of the baseline EMG) in the MS patients was comparable to normal values, but the transcallosal conduction time was significantly delayed (patients 17.2 +/- 6.4 ms; normal subjects 12.2 +/- 2.6 ms; P < 0.001). The duration of the inhibition was significantly prolonged in MS patients (patients 47.9 +/- 20.9 ms; normal subjects 38.9 +/- 10.1 ms; P = 0.02). Transcallosal conduction time was delayed in 11 (46%) of 24 patients, compared with normal subjects. It exceeded the normal range (mean +/- 2.5 SD) in one normal subject (specificity 96%). No correlation could be found between the size or extent of the lesions obtained from the MRI scan and the onset latency or the depth of the inhibition. CONCLUSIONS: We conclude that conduction over transcallosal connections is significantly slower in patients with MS. PMID- 9741790 TI - Modeling direct activation of corticospinal axons using transcranial electrical stimulation. AB - Corticospinal axons can be directly activated using anodal transcranial electrical stimulation. The purpose of this work was to find the location of the direct activation. The response to stimulation was modeled with a spherical head model and an active model of a corticospinal nerve. The nerve model had a deep bend at a location corresponding to a corticospinal fiber entering the midbrain. The threshold activation initiated close to brain surface; the exact location depended on whether the cell body located in the surface layers of the brain or in the bank of the central sulcus. The stimulation time constant was 44 micros. When the stimulus amplitude was increased, the site of activation shifted gradually to deeper level, until the activation initiated directly at the bend causing a half millisecond latency jump at spinal level. These results support the theory that the corticospinal axons can be directly activated at deep locations using anodal transcranial electrical stimulation. However, the high amplitude needed for the direct activation suggests that not only the bends on the fibers, but also the shape of surrounding volume conductor (intracranial cavity) favor activation at this location. PMID- 9741791 TI - Discharge pattern of human motor units during dynamic concentric and eccentric contractions. AB - OBJECTIVES: A total of 45 motor units (MUs) from the human biceps brachii muscle were investigated during isovelocity concentric and eccentric movements performed by means of a device implementing an external torque in the direction of the extension proportionally to the elbow angle changes. The effects of movement velocity on the recruitment and decruitment thresholds (RT and DT) and the corresponding discharge patterns were determined. METHODS: A wire branched electrode placed subcutaneously was used to discriminate the potentials from a single MV. RESULTS: The majority of MUs (91%) were recruited at lower torque values with the increase of movement velocity. The decrease of RT was statistically significant for 47% of the investigated MUs. A typical discharge pattern of short first interspike interval (ISI) followed by a longer one was observed for 93% of all MUs. After the first 2-3 spikes the rate of the MU discharge was approximately constant regardless of the fact that the muscle force gradually increased until the end of the concentric movement. CONCLUSIONS: There are differences in the muscle force control during shortening and lengthening contractions. For 82% of the investigated MUs DT was smaller at faster movements and for 21 MUs (47%) the decrease of DT was significant. The gradually declined MU discharge rate throughout the entire movement with a very long last ISI was demonstrated for 93% of the investigated MUs. PMID- 9741792 TI - EMG fatigue patterns accompanying isometric fatiguing knee-extensions are different in mono- and bi-articular muscles. AB - OBJECTIVES AND METHODS: Isometric, fatiguing knee-extensions at 30%, 50% and 70% maximum voluntary contraction (MVC) were performed by 18 healthy human subjects. Surface electromyographic (SEMG) activity was recorded from the mono-articular vastus medialis (VM) and vastus lateralis (VL) muscles, and the bi-articular rectus femoris muscle (RF). To make the bi-articular muscle work under (1) constant and (2) similar working conditions as the two mono-articulars do, the hip was fixed in a flexed position. The root mean square (RMS) SEMG recorded during fatigue was standardized to the respective values of MVC. The mean coefficients of regression of the RMS and median frequency (MF) changes were then analyzed by multivariate analysis of variance. RESULTS: The load effect upon the muscle fatigue changes, as measured by increase in RMS EMG, differed between the bi-articular muscle and the two mono-articulars, in that the parameter dropped with maximum load for the bi-articular, whilst it remained stable or even increased for the mono-articulars. This might suggest that the mono- and bi articular muscles have different roles in fatigue tasks where the bi-articulars function purely as mono-articulars. By contrast, such a clear dichotomy between the bi-articular RF and the two mono-articulars, VM and VL, was lacking for the fatigue parameter of MF. CONCLUSIONS: As these findings were confined to the changes in RMS EMG, different neuronal coding mechanisms for the mono- and bi articular muscles in the central nervous system may be inferred. PMID- 9741793 TI - Increased excitability of the human corticospinal system with hyperventilation. AB - OBJECTIVES: Hyperventilation is effective in inducing generalized spike-wave discharges in patients with absence seizures and improves visual function and normalizes visual function in patients with multiple sclerosis. Hyperventilation increases the excitability of cutaneous and motor axons. In experimental animals, hyperventilation increases excitability of hippocampal neurons. There is however no direct evidence of a hyperventilation-induced increase in neuronal excitability within the central nervous system in humans. In this study we determined the effects of hyperventilation on the human corticospinal system. METHODS: We studied the effects of hyperventilation on (1) motor evoked potentials (MEPs) induced by transcranial magnetic pulse stimulation of the motor cortex and (2) F-wave responses. Six subjects were studied. RESULTS: Hyperventilation resulting in an end-tidal pCO2 of 15 mm Hg or less enhanced the amplitude of the MEP and resulted in a shortened onset latency. F-wave amplitudes were enhanced without any change in onset latency. CONCLUSIONS: These findings indicate that hyperventilation increases the excitability of the human corticospinal system. A hyperventilation-induced increase in excitability within the central nervous system may account for clinical phenomena such as facilitation of spike-wave discharges. PMID- 9741794 TI - Median mixed and sensory nerve conduction studies in carpal tunnel syndrome. AB - OBJECTIVE: To assess the sensitivities and specificities of velocity differences between median mixed nerve conduction across the wrist (Medmxpw) and (I) median mixed nerve conduction in the forearm (Medmxf) and (II) palm to D2 sensory conduction (MedpD2). DESIGN AND METHODS: We prospectively studied 67 limbs of patients with clinically definite carpal tunnel syndrome (CTS). Medmxf and Medmxpw were performed by stimulating the median nerve at the elbow and palm respectively and recording at the proximal wrist crease. We also compared conventional median sensory (D2-wrist) and mixed (palm-wrist) tests in all patients. Thirty limbs of asymptomatic subjects served as normal controls and 21 limbs of subjects with other neuropathies served as diseased controls; control data was collected prospectively. RESULTS: The sensitivity of the MedpD2-Medmxpw difference (0.87) was significantly greater than that of the Medmxf-Medmxpw difference (0.61, P < 0.001). Both tests were similar and highly specific (0.98 and 0.96, respectively). CONCLUSIONS: The MedpD2-Medmxpw study is among the most sensitive and specific electrophysiologic tests for CTS. PMID- 9741796 TI - Magnetic stimulation mapping of motor cortex: factors contributing to map area. AB - Transcranial magnetic stimulation (TMS) can evoke an electromyographic response in muscles of the hand with stimulation at a large number of scalp sites widely spaced over the contralateral primary motor area of the brain. To determine the extent to which this is due to current spread from the stimulating coil to a smaller region of excited cortex, excitability curves of motor evoked potential amplitude vs. stimulus intensity were measured at multiple scalp sites. It was found that these curves were of very similar shape, but with different offsets along the stimulus intensity axis. This could be explained on the basis of current flow to a small excitable region of cortex located at some depth in the brain. It is concluded that the surface area of TMS maps is primarily determined by current spread and by the relationship between the position of the coil on the scalp and the depth of the motor output region in the cortex, and does not necessarily provide a true indication of the spatial extent of the motor output region projecting to the target muscle. PMID- 9741795 TI - Subclinical neuropathy in type I diabetic children. AB - OBJECTIVES: Small and large, somatic and autonomic nerve fibre functions were neurophysiologically evaluated in 33 asymptomatic neurologically free type I diabetic children and 69 age-matched healthy controls. METHODS: The evaluation of large and small somatic nerve fibre function was performed by conventional nerve conduction studies, thermal specific and thermal pain sensitivity tests, as well as autonomic nerve fibre functions by sympathetic skin response and R-R interval variation assessment. RESULTS: A significant difference was established between the healthy and the diabetic group. Neurophysiologically determined subclinical neuropathy was found in 87% of type I diabetic children. The majority of abnormal recordings were found on the lower limbs. The dysfunction of the somatic motor large nerve fibre type in the lower limbs was altered in 57% of patients, somatic sensory large in 39%, somatic sensory small in 45%, and sympathetic in 45%. The leading abnormal measure was a delayed sympathetic skin response on the foot (42% of diabetic children) followed by a reduced amplitude of sural nerve action potential (36%). The whole spectrum of recordings showed scattered involvement of nerve functions. There was no selective susceptibility of nerve fibre types exposed to a noxious factor. CONCLUSION: A complex neurophysiological assessment, including standard nerve conduction studies as well as psychophysical examination and autonomic nerve function tests, evaluating the function of small and large nerve fibres, is recommended for evaluating the subclinical neuropathy in asymptomatic type I diabetic children. PMID- 9741797 TI - Representation of cortical motor function as revealed by stereotactic transcranial magnetic stimulation. AB - Cortical motor representation of 12 muscles of the trunk and the upper and lower extremity was investigated in 18 healthy subjects using focal transcranial magnetic stimulation (TMS) in conjunction with a frameless stereotactic system (FSS). This combination allowed us to orientate stimulation sites to the individual central sulcus rather than to bony landmarks. Distinct but overlapping areas of muscle representation were identified and the 3-dimensional representation of those 12 muscles along the course of the central sulcus was obtained. With increasing stimulus intensity, the cortical output maps changed in that more muscles became excitable, motor evoked potential (MEP) amplitude and size of the responsive area increased and latency of the MEP decreased. These effects were more pronounced for proximal than for distal muscles, indicating a more widespread organization of corticospinal motor projection related to proximal muscles. The combination of TMS and FSS represents a method with which functional information can be directly related to underlying cortical anatomy. This correlation will be useful in the assessment of higher brain functions with TMS. PMID- 9741798 TI - Magnetic stimulation over different brain regions: no differential effects on the elicited sympathetic skin responses. AB - Peak latencies and amplitudes of sympathetic skin responses (SSRs) of the hand following magnetic stimulation at different sites with two or five consecutive 10 Hz stimuli were investigated with regard to safety aspects of repetitive transcranial magnetic cortex stimulation (rTMS). The amount of sympathetic activation as assessed by the amplitudes of the SSRs depended on the stimulation site and decreased in the following order: brachial plexus stimulation > nerve root stimulation > stimulation over the brain > activation by acoustic coil artefact. When stimulating over six different regions of the cortex (frontal, central, parieto-occipital, and both hemispheres), the elicited SSRs had similar amplitudes and peak latencies. The SSRs elicited by rTMS over the motor cortex were not related to the sum of the amplitudes of excitatory muscle compound responses. Currents with opposite directions over the motor cortex markedly influenced the size of the motor responses but not of the SSRs. The number of consecutive 10-Hz stimuli did not influence the latencies or amplitudes of the SSRs. It can be concluded that SSRs after magnetic stimulation over peripheral nerves or the brain are a correlate of an unspecific arousal reaction. A therapeutic application of short series of rTMS should not be limited by the amount of sympathetic activation. PMID- 9741799 TI - Characteristic response to transcranial magnetic stimulation in Rett syndrome. AB - To pathophysiologically evaluate the corticospinal tracts (CSTs) in Rett syndrome (RS), transcranial magnetic stimulation (TMS) was performed in 3 patients aged 4, 6 and 13 years. The two younger cases exhibited the clinical characteristics of the pseudostationary stage (stage III), while ambulation was lost in the oldest case at the age of 11 years (stage IV). The motor cortex and cervical spinal roots were magnetically stimulated to obtain motor evoked potentials (MEPs) from the relaxed first dorsal interosseous muscle. Compared with the central motor conduction time (CMCT) in age-matched normal children, CMCT in the stage III cases was significantly short (6.9-7.1 ms, P < 0.05). In the stage IV case, CMCT was markedly short but not significantly so (6.6 ms, P = 0.06), which was partly due to a significant increase in the threshold intensity of TMS (100%, P < 0.05). Thus, the CMCT shortening, which implied unique cortical hyperexcitability, was considered consistent and characteristic of RS. The CSTs in the stage IV case were certainly impaired, corresponding well to the progressive spastic paresis. PMID- 9741800 TI - Variability in the amplitude of skeletal muscle responses to magnetic stimulation of the motor cortex in man. AB - We have investigated variability in the amplitude of compound motor evoked potentials (cMEPs) in right and left thenar and wrist extensor muscles in response to synchronous bilateral transcranial magnetic stimulation (TMS) of the motor cortices using two figure-of-eight stimulating coils. Trials of 50 stimuli revealed a wide range of variability in cMEP amplitudes in relaxed muscles (coefficient of variation, range 0.22-1.12). The amplitudes of the cMEPs in one muscle correlated positively with those in the others. The r2 values (mean +/- SEM) were 0.27 +/- 0.06 for muscles on the same side of the body and 0.19 +/- 0.04 for muscles on opposite sides. Employing the ECG to trigger TMS, clamping the coil relative to the head or altering the orientation of the coil all failed to affect the variability of cMEPs. We conclude that fluctuations in excitability of the corticospinal pathway give rise to the variability in the response to TMS, that they are wide-ranging with respect to the muscles affected, and include a bilateral component. We argue that the variability reveals fluctuations in excitability mainly at the cortical rather than the spinal level. We suggest that measures of variability might provide an indication of cortical activity in conditions where consciousness and voluntary movement are compromised. PMID- 9741801 TI - A simple method for recording motor evoked potentials of lingual muscles to transcranial magnetic and peripheral electrical stimulation. AB - Motor evoked potentials were recorded from lingual muscles by means of clip electrodes applied on the lateral side of the tongue, following transcranial magnetic stimulation and peripheral electrical stimulation of the 12th cranial nerve at the mandible jaw. Using a circular coil, the stimulation of the cerebral cortex elicited a response of about 8 ms: its amplitude was higher in the right tongue placing the coil over the contralateral hemisphere, 4 cm from the vertex, with coil currents flowing counterclockwise. Coil position and current flow direction did not significantly modify the characteristics of responses recorded from the left side. The separate stimulation of either hemisphere was better obtained using an 8-shaped coil. The latency of the motor response measured 7.7 8.0 ms, the amplitude was greater on stimulation of the contralateral than the ipsilateral hemisphere and was larger recording from the right (3.3 +/- 1.1 mV) than from the left (1.2 +/- 0.7 mV) side. Positioning the circular coil over the parieto-occipital skull, a response of 4.1 +/- 0.3 ms was obtained, reflecting the intracranial activation of the hypoglossal nerve. The peripheral stimulation at the mandible elicited a response of 3.2 +/- 0.5 ms. The method described appears simple and reliable, potentially helpful in clinical practice. PMID- 9741802 TI - A data dependent computer algorithm for the detection of muscle activity onset and offset from EMG recordings. AB - This paper describes modifications to an algorithm presented by Marple-Horvat and Gilbey (1992) for identifying bursts of muscle activity in electromyographical (EMG) recordings. Our efforts to apply their algorithm to spontaneously moving infants and toddlers resulted in limited success. The modified algorithm makes several parameters dependent on the data being analyzed; these changes enabled it to analyze a variety of EMG recordings more effectively. The original algorithm had a success rate (correctly identified bursts) of 62.9% and combined error rate (number of insertions and deletions) of 73.0% when applied to an independent test data set. The modified algorithm displayed a success rate of 85.4% and combined error rate of 23.6%. PMID- 9741803 TI - Concentric needle recording characteristics related to depth of tissue penetration. AB - This study investigates the influence of tissue penetration depth as it relates to a concentric needle electrode, particularly delineating regions where the cannula potential predominates over the core potential. The regions of cannula predominance is studied by means of a standard and 20 times enlarged physical model of an electromyographic concentric needle electrode in a homogeneous volume conductor by delineating the zero isopotential which partitions where the core potential predominates versus where the cannula potential predominates. Clinical studies in muscle tissue are used to test and confirm results from the enlarged physical model. At shallow electrode insertions equivalent to 4 mm, the concentric needle model records a net negative potential, which is a region where the cannula predominates, from a distant positive dipole at the same depth compared with a net positive potential for penetration depths exceeding 4 mm. The clinical portion of this study verifies the bipolar nature of the concentric needle electrode in detecting motor unit action potentials (MUAPs) with primarily an initial positive onset irrespective of recording depth. Refinements to the conceptualization of the nature and detection of MUAPs are discussed which are consistent with all the findings of the clinical and model study. PMID- 9741804 TI - Locomotor training in paraplegic patients: a new approach to assess changes in leg muscle EMG patterns. AB - This study describes an amplitude independent assessment of changes in leg muscle EMG patterns in both complete and incomplete paraplegic patients during the course of locomotor training. The approach expresses the change as an approximation of the patients' gait EMG pattern compared with that of healthy subjects. The variation ratio (VR), coefficient of variation (CV) and Pearson's correlation coefficient (R), are used as measures of the dissimilarity/similarity of a set of wave forms. These parameters were evaluated for their ability to assess changes in the EMG pattern of the patients with respect to that of healthy subjects. The VR showed the best correlation to our data and was therefore considered to represent the optimum variable in the assessment of changes in EMG patterns. PMID- 9741805 TI - Locomotor capacity and recovery of spinal cord function in paraplegic patients: a clinical and electrophysiological evaluation. AB - Recent studies have shown that a locomotor pattern can be induced and trained into paraplegic patients under conditions of body unloading using a moving treadmill. The present study investigated the behaviour of the locomotor pattern and also the relationship of its development to the spontaneous recovery of spinal cord function assessed by clinical and electrophysiological (tibial nerve somatosensory evoked potentials and motor evoked potentials) examinations. The earliest time that spinal locomotor activity could be induced was when signs of spinal shock had disappeared. This activity was distinct from spinal stretch reflex activity. In complete and incomplete paraplegic patients an increase of gastrocnemius electromyographic activity occurred during the stance phase of a step cycle with daily locomotor training over the whole training period of 12 weeks. This was coincident with a significant decrease in body unloading. In contrast to this, neither clinical nor electrophysiological examination scores improved after the onset of training in both patient groups. Only in incomplete paraplegic patients was there an insignificant increase in sensory and motor scores obtained in the neurological examination during the time period before onset of training. An improvement of locomotor function by training was also seen in patients with paraplegia due to a cauda lesion. Therefore, in patients with a spinal cord lesion training effects on muscles and tendons are present in addition to those on the spinal locomotor centres. The findings of this study may be relevant for future clinical treatment of paraplegic patients. PMID- 9741806 TI - Event-related beta synchronization after wrist, finger and thumb movement. AB - Pre-movement event-related desynchronization (ERD) and post-movement event related synchronization (ERS) were studied in a group of normal subjects during voluntary thumb, index finger and wrist movement. The band power time courses were computed for the upper alpha band (10-12 Hz) and for two frequency bands in the range of beta (16-20 Hz and 20-24 Hz). While a similar mu ERD was found during motor preparation for the 3 movement tasks, significant differences concerning beta synchronization were observed after movement off set. The contralateral percentage beta increase (ERS) was significantly larger in gross movements of the wrist as compared to index finger and thumb movements, which is discussed under the assumption of a cumulative effect. Summarizing, pre-movement desynchronization seems relatively independent of the forthcoming type of movement, whereas the post-movement beta synchronization might depend on the activated muscle mass. PMID- 9741807 TI - Influence of chronic administration of L-DOPA on event-related desynchronization of mu rhythm preceding voluntary movement in Parkinson's disease. AB - The spatiotemporal pattern of event-related desynchronization (ERD) during the motor preparation period preceding a self-paced voluntary wrist-flexion was compared in two groups of 10 right and 10 left hemiparkinsonian patients, before and after chronic administration of L-DOPA. ERD was computed in the 9-11 Hz frequency band from 11 source derivations covering the medial frontocentral, central and parietocentral areas, during two successive left and right experimental conditions (100 self-paced wrist flexions). In the two groups ERD appeared with a shorter latency over the contralateral primary sensorimotor area, when the movements were performed with the akinetic hand. After L-DOPA administration, earlier ERD onset before the movement was observed in both groups over the contralateral and ipsilateral central and parietocentral areas. A medial frontocentral ERD distribution was also observed before the onset of movement, especially in the right hemiparkinsonian group. Delayed ERD onset, which shows that programming of movement is affected in Parkinson's disease, may be partially corrected by L-DOPA therapy. PMID- 9741808 TI - Effects of tactile interference stimulation on somatosensory evoked magnetic fields following tibial nerve stimulation. AB - We studied the effects of interfering tactile stimulation applied to the foot ipsilateral and contralateral to the stimulation on somatosensory evoked magnetic fields (SEFs) following tibial nerve stimulation at the ankle. Equivalent current dipoles (ECDs) of all 4 components, 1M-4M, in all sessions were estimated to be very close each other, around the foot area of the primary sensory cortex (SI). The 1M, 2M and 4M components were significantly reduced in amplitude by the ipsilateral-foot interference, and we consider that this phenomenon is due mainly to 'saturation' of the neurons in area 3b of the SI. In contrast, the 3M component was significantly enhanced in amplitude by the contralateral-foot interference. We suspect that this result was due to the effects of neuronal activities in areas 2, 5 and/or 7, which receive inputs from both sides of the body, i.e. to 'bilateral function'. Considering the various types of interference effects on SEFs and somatosensory evoked potentials (SEPs) observed in not only the present, but also in the previous studies, we conclude that both SEFs and SEPs following tibial nerve stimulation are generated mainly by ascending signals mediated by cutaneous fibers of the peripheral nerves rather than the muscle afferents. PMID- 9741809 TI - Waveform and habituation of sympathetic skin response. AB - The aim of this study was to investigate the relation of sympathetic skin response (SSR) waveform variation to latency, amplitude, and habituation. Twenty SSRs were recorded from the palm skins of 50 normal subjects by electrical stimulations. Reproducibility of SSR waveforms was also evaluated (n = 35). Waveforms were classified as one of two types, namely, the P type, in which the positive component was larger than the negative, and the N type, in which the negative component was larger than the positive. During successive stimulations, 14 of the 50 subjects had only P type waveforms (P pattern), 9 others only the N type (N pattern), and the remaining 27 both the P and N types (M pattern). The P pattern had a larger amplitude and shorter latency than the N pattern. Twenty seven of the 35 subjects had the same SSR pattern in two tests conducted on different days. Habituation with time was more pronounced for the M pattern than for the other two patterns. It was suggested that changes in SSR waveform during successive stimulations reflected habituation. Most subjects with the M pattern had a distinct attribute in the development of habituation. PMID- 9741810 TI - Eye movement abnormalities in myotonic dystrophy. AB - We studied saccade and smooth pursuit eye movements in 31 patients suffering from myotonic dystrophy (MD). On the basis of mean value comparisons, saccades were slower and hypometric and smooth pursuit eye movements performed worse in MD patients than in controls. On an individual basis, saccade duration was prolonged in 67.7%, saccades were hypometric in 19.4%, saccade latency was delayed in 9.7%, and the smooth pursuit performance index was decreased in 9.7% of patients. Eye movement abnormalities did not correlate with those detectable by visual, brain stem auditory and somatosensory evoked potentials. We attempted to classify eye movement abnormalities as myogenic or neurogenic on the basis of differences in combination of eye movement abnormalities and the occurrence of D5/D35 dissociation; the latter consists of a prolonged duration for large (35 degrees) but not for small (5 degrees) saccades. Since D5/D35 dissociation occurred in 26/33 multiple sclerosis patients with increased saccade duration, we considered it to be a neurogenic pattern attributable to a central nervous system (CNS) dysfunction. A prolonged duration without dissociation especially in combination with saccade hypometria, is interpreted as a myogenic pattern, although the lack of dissociation may also occur with CNS impairment in case of a marked increase in saccade duration. Accordingly we classified the oculomotor abnormalities detected as neurogenic in 11 MD patients and as myogenic in another 10, but in some subjects belonging to the second group concomitant CNS impairment is not to be excluded. PMID- 9741811 TI - A case of Guyon syndrome with neuroapraxic block resolved after surgical decompression. AB - Guyon syndrome is the well-known ulnar entrapment at the wrist; usually surgical decompression improves the symptoms. Neurophysiological studies are essential to demonstrate the abnormality of ulnar nerve conduction at the wrist. We report a case of Guyon syndrome onset after 6 h of cycling. Neurophysiological study revealed ulnar neuroapraxic block at the wrist, and axonal impairment. Post operative clinical and neurophysiological follow-up showed marked clinical improvement and neurophysiological resolution of abnormalities. To our knowledge, no cases of Guyon syndrome with neuroapraxic block are reported in the literature. PMID- 9741812 TI - A custom designed system to measure corticospinal tract jitter. AB - Typical latency of an individual limb muscle response to magnetic or electric stimulation of the human cortex is in the range of 10-50 ms. For the latency variability, i.e., jitter studies, a resolution of at least 20 micros is needed. Commercially available EMG equipment needs custom-designed upgrading to allow for such studies. Two solutions were designed: (i) a hardware unit allowing an adjustable delay of data acquisition after the delivered stimuli; and (ii) diverting of the amplified biological signal and the EMG equipment trigger to an external computer equipped with an analogue-to-digital conversion (ADC) module. Custom-designed software made fast ADC possible during the whole period of data acquisition. Both concepts were applied to a Vickers Medical Mystro electromyograph, and have been successfully used in the Ljubljana (Slovenia) Institute of Clinical Neurophysiology for the last 2 years. PMID- 9741813 TI - Is it possible to create a perfect external control system? PMID- 9741814 TI - Connective tissue markers of rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is a common systemic autoimmune disorder of unknown aetiology. The most common outcome of RA is a progressive development of joint destruction and deformity. Early introduction of disease-modifying antirheumatic drugs seems important for prevention of the long term injuries of articular cartilage and bone. Early diagnosis and selection of patients with rapidly progressive disease therefore is of clinical significance. Routine laboratory tests are valuable in monitoring for renal, hepatic and haematological side effects of medical treatment. Determination of rheumatoid factor contributes to the classification of arthritis as RA, and acute phase reactants are useful for quantifying and comparing the level of inflammatory activity in the course of a given patient. There is, however, a lack of sensitive and specific biochemical markers for RA, and frontline biochemical research is devoted to characterizing molecules which are of diagnostic and prognostic value, as well as molecules which are indicators of the degree of joint cartilage and bone destruction. The present survey summarizes current knowledge concerning possible tissue-specific marker molecules of RA. PMID- 9741815 TI - Platelet adhesion in patients prone to arterial and venous thrombosis: the impact of gender, smoking and heredity. AB - Platelet adhesion was measured in 271 consecutive subjects (151F, 120M) referred to the department for investigation of their propensity to develop thrombosis. Arterial thrombosis was the cause in 27% of the subjects and pulmonary embolism in 23%, whereas venous thrombosis was the cause in 50%. Ninety-three patients were using nicotine in the form of smoking or snuffing, 45 were ex-users and 130 patients never-users. Adhesion was measured as the retention of platelets in a commercially available column of glass beads. After strict standardization of the method the reproducibilities within-day and between-day were good. Platelet retention was increased in thrombosis-prone patients as compared to references (p=0.016). This increase was seen irrespective of type of thrombosis. Multifactor ANOVA analysis revealed a strong dependence of gender and smoking habits with higher platelet retention in men and in ex-smokers and current smokers. The highest levels were found in ex-smokers with arterial thrombosis and in current smokers with pulmonary embolism. In the control population we found high platelet retention in smokers (p=0.001) and in those with a family history of thrombosis (p=0.0025). It is concluded that the measurement of platelet retention may form a basis for the selection of patients to antiplatelet therapy and that the activity of platelets is affected by smoking and related to sex and family history of thrombosis. It is also concluded that thrombus formation in men and women may be governed partly by different mechanisms. PMID- 9741816 TI - Pig epidermal growth factor precursor contains segments that are highly conserved among species. AB - The 53-aa polypeptide epidermal growth factor (EGF) is synthesized as a 1200-aa precursor. The non-EGF part of the precursor is very long compared with EGF, and can therefore be expected to have a biological role of its own. We have sequenced cDNA of the pig EGF precursor and compared a 668-aa segment with that of the human, the rat and the mouse EGF precursors, in order to identify highly conserved domains. The examined part of the precursor contains EGF itself and six so-called EGF-like modules. The overall amino acid identity among the four species is 64%. However, the amino acid identity differed from around 30% in some segments to around 70% in others. The highest amino acid identity, 71%, was observed for a 345-aa segment that contains three EGF-like modules and which is homologous to a part of the low-density lipoprotein receptor (LDL receptor). The amino acid identities are 64% for EGF itself, and 50-67% for the remaining three EGF-like modules. The segment of the LDL receptor that is homologous to a part of the EGF precursor is important for the function of the LDL receptor, and EGF-like modules seem to be involved in protein-protein interactions in a number of proteins. In conclusion, some segments of the EGF precursor are remarkably well conserved among species and it is tempting to speculate that they have a biological function. PMID- 9741817 TI - Quantification of mRNA levels of endothelin receptor subtypes and preproEndothelin-1 in renal needle biopsies by competitive reverse transcriptase polymerase chain reaction. AB - The vasoconstrictive peptide endothelin-1 (ET-1) is an autocrine/paracrine peptide of putative pathophysiological importance in renal transplant medicine. The aim of the present study was to develop a method for analysis of gene expression of the renal endothelin system in humans. Only small amounts of tissue are available from renal cortical needle biopsies. Thus, in the present study we developed a quantitative assay based on the competitive reverse transcriptase polymerase chain reaction (RT-PCR) technology. We quantified endothelin A (ET(A)) and B (ET(B)) receptor subtype mRNAs and preproET-1 mRNA levels in renal cortex biopsies obtained before nephrectomy of healthy kidney donors. Mean (+/- SEM) mRNA levels of the ET(A) and ET(B) receptor subtypes in 26 living donors were 212 +/- 23 and 368 +/- 56 amol/microg total RNA, respectively. The preproET-1 mRNA level in 19 living donors was 213 +/- 28 amol/microg total RNA. The inter-assay coefficient of variation (CV) for the assay was 10%; the intra-assay CV was 6 13%. The competitive RT-PCR assay described provides an accurate tool for gene expression investigation of the human endothelin system in renal cortical needle biopsies. PMID- 9741818 TI - Clinical evaluation of a diagnostic strategy for deep venous thrombosis with exclusion by low plasma levels of fibrin degradation product D-dimer. AB - Clinical research studies have indicated the possibility of diagnostic strategies for deep venous thrombosis (DVT), strategies which include a step where the diagnosis is excluded by low or undetectable plasma levels of fibrin degradation product D-dimer. In collaboration with two local hospitals in Sweden, three implementations of such a strategy are evaluated in this study. Procedures 1, 2 and 3 differed in the method for D-dimer determination, i.e. latex agglutination, immunofiltration and both, respectively. The evaluated procedures were performed in parallel and compared with the current procedure in the different hospitals. At both hospitals, the current procedure stipulated mandatory phlebography and laboratory analysis of acute coagulation status and routine haematology with report-back time of 2 h. Within the 2 h the hospitals' clinical chemistry laboratories also determined plasma D-dimer by the two methods. Of 180 patients enrolled in the study, phlebography was successful in 155 and unsuccessful in 25. The phlebographies revealed 47 proximal DVT, 13 distal DVT and 95 no DVT. With Procedure 1, 53 patients (29%) were excluded in the D-dimer step. For these patients, 47 successful phlebographies revealed one proximal DVT and two distal DVT. With Procedure 2, 71 patients (39%) were excluded. For these patients, 65 successful phlebographies revealed two proximal DVT and four distal DVT. With Procedure 3, 44 patients (24%) were excluded. For these patients, 41 successful phlebographies revealed two distal DVT. The negative predictive values of the D dimer exclusion step, with 95% confidence intervals given within parentheses, were 96% (88-100%), 91% (84-98%) and 95% (89-100%) for Procedures 1, 2 and 3, respectively. The evaluation demonstrated that the diagnostic potential of D dimer revealed in research studies can be achieved in clinical practice. The study also indicated that the positive diagnostic value of high levels of D-dimer may be of use in finalizing the diagnosis in the 14% of patients for whom phlebography is unsuccessful. PMID- 9741819 TI - Monitoring erythrocyte free radical resistance in neonatal blood microsamples using a peroxyl radical-mediated haemolysis test. AB - Inadequate resistance to oxidative stress has been implicated in several diseases of premature children. Antioxidative defences at the membrane level can be studied by measuring haemolysis induced through exposure of erythrocytes to the free radical generator AAPH (2,2'-azobis (2-amidinopropane)dihydrochloride). We developed a micromodification of this haemolysis test requiring only 15 microl of erythrocytes derived from capillary blood samples. The time needed for 50% haemolysis (T50%) was used to characterize radical resistance of erythrocytes. T50% results in adult samples were highly reproducible. T50% values in healthy term infants on the first 2 d of life were lower than in adults (p < 0.001), but increased to the same level thereafter. A correlation was found between T50% values and plasma tocopherol levels as determined in plasma of each of the capillary blood samples (p < 0.001). On the first day of life T50% results in preterm infants (n = 20) were higher than in term infants (p < 0.001). It was easy to monitor T50% results and plasma tocopherol levels in preterm infants that were not at all burdened by the sampling method, almost daily over several weeks. The micromodification presented simplifies monitoring of antioxidative defences in sick preterm infants. PMID- 9741820 TI - Association of factor VII protein concentration with lifestyle factors. AB - Several studies have demonstrated that changes in fasting levels of factor VII coagulant activity (FVII:C), a thrombotic risk marker, are due to changes in FVII protein concentrations (FVII:Ag). Consequently, studies on FVII now often include measurements of FVII:Ag. The present cross-sectional study examined the association between several behavioural variables (body mass index, physical activity, tobacco or alcohol consumption) or physiological variables (total cholesterol, HDL-cholesterol, triglycerides, glucose, insulin, fibrinogen, resting pulse, systolic blood pressure, bleeding time) and FVII:Ag in 439 51-y old Danish men. In the multivariate analyses, body mass index (BMI), low physical activity, total cholesterol, short bleeding time, and insulin showed an independent positive association with FVII:Ag. The strongest independent association with FVII:Ag was found for total cholesterol. These results suggest that blood lipids are major determinants of FVII:Ag, but that other lifestyle factors such as insulin, BMI and physical activity can also influence FVII:Ag. Furthermore, the association between FVII:Ag and bleeding time suggests an effect of FVII or the FVII pathway on primary haemostasis. PMID- 9741821 TI - Serial measurements of cardiac markers to rule in or out acute myocardial damage less than 3 h after admission in acute chest pain patients without ECG-signs of acute myocardial infarction. AB - Acute chest pain patients without ECG-signs of acute myocardial infarction (AMI) on admission need to be earlier and better diagnosed to reduce use of expensive intensive care beds and to treat more patients with acute recirculation therapy. We investigated whether total CK-activity, CK-MB mass, CK-MB2, myoglobin, cardiac troponin I (cTnI) and T (cTnT) measured in venous blood on admission and after 1 and 2 h could be used to identify or exclude acute myocardial damage (AMD) in 22 acute chest pain patients without ECG-signs of AMI admitted to hospital within 6 h after onset of pain. Increases in CK-MB mass, CK-MB2, myoglobin and cTnI identified AMD in three patients classified retrospectively as AMI. Likewise, CK MB mass, CK-MB2, cTnI and cTnT increased with time in three of seven patients classified as having unstable angina pectoris. CK-MB2 and cTnI increased with time in two patients with tachycardia belonging to the other heart disease group. The remaining seven patients of the non-heart disease group showed no change in any of the cardiac markers. Thus, early serial measurements of selected cardiac markers appear useful in identifying or excluding AMD 3 h after admission in these acute chest pain patients. PMID- 9741822 TI - Urinary excretion of catecholamines in hospitalized and non-hospitalized healthy children and adolescents. AB - Non-conjugated catecholamines were measured in morning urine samples from 111 healthy, non-hospitalized subjects aged 8-18 y and in 16 hospitalized, healthy subjects aged 12 16 y. The catecholamines were extracted by cation exchange columns and alumina adsorption and quantitated with HPLC with electrochemical detection. The concentration of catecholamines was related both to the urinary creatinine concentration and to the collecting period and body surface area. Linear regression analysis was used to estimate continuous age-related reference centiles based upon the measurements from the 111 non-hospitalized subjects. The upper limits for the adrenaline/creatinine and noradrenaline/creatinine ratios were lower than in previous studies. The excretion of adrenaline and noradrenaline per hour and m2 body surface area was higher in the 16 hospitalized than in the 74 age-matched non-hospitalized subjects. The excretion of the catecholamines expressed per hour and m2 body surface area and expressed relative to creatinine excretion, decreased with increasing age, and the excretion of adrenaline and noradrenaline per hour and m2 body surface area was higher in boys than in girls. In conclusion, standardization of urine sampling leads to more narrow ranges for urinary adrenaline and noradrenaline excretion in healthy children and adolescents. PMID- 9741823 TI - Thrombin signal transduction of the fibrinolytic system in human adult venous endothelium in vitro. AB - Thrombin can regulate the-fibrinolytic system by increasing the endothelial production of both tissue plasminogen activator (t-PA) and plasminogen activator inhibitor type-1 (PAI-1). The thrombin receptor transducts signals through the GTP-binding protein system, the classical pathway being the Galpha q-protein. The purpose of the present study was to examine the roles of Galpha i-protein and tyrosine kinases in the thrombin signal transduction of t-PA and PAI-1 production from human adult vein endothelial cells (HAVEC). t-PA and PAI-1 antigen were analysed in conditioned medium from cultured HAVEC after 16 h incubation. Data are expressed as percentages of basal release (100%), means +/- 95% confidence intervals. Thrombin increased t-PA and PAI-1 production (234 +/- 42% and 211 +/- 42%, respectively). Pertussis toxin (PTX) (inhibiting Galpha i-pathway) reduced basal PAI-1 (66 +/- 8%), but had only a weak influence on basal t-PA production. Pertussis toxin and genistein (inhibiting tyrosine kinase) significantly reduced the thrombin induction of both t-PA and PAI-1 (PTX: 142 +/- 23% and 146 +/- 19%, respectively, genistein: 156 +/- 42% and 76 +/- 24%, respectively). The present study demonstrated that thrombin can increase the production of t-PA and PAI-1 by transducting signals through the Galpha i and tyrosine kinase pathway, in addition to the Galpha q/protein kinase C pathway as has been found previously. PMID- 9741824 TI - Insulin-like growth factor-I and insulin-like growth factor binding protein-1 in a representative population of type 2 diabetic patients in Sweden. AB - OBJECTIVE: To study the influence of type 2 diabetes on the insulin-like growth factor-I (IGF-I) and insulin-like growth factor binding protein-1 (IGFBP-1) serum levels in an area-based population of type 2 patients previously described. RESULTS: The patients (n = 151) were elderly (70.6 +/- 0.7 years of age) and moderately overweight (BMI 27.0 +/- 0.4 kg/m2). Most patients (83%) were treated with either diet alone or diet in combination with sulphonylurea. Metabolic control measured as HbAlc deteriorated with duration (p < 0.001) and between groups treated with diet (HbA1c 5.8 +/- 0.6), sulphonylurea (7.5 +/- 0.2) and insulin (7.7 +/- 0.4). Mean levels of IGF-I were within reported normal range, but were lower in the insulin-treated as compared to the non-insulin-treated patients. Levels of IGF-I decreased with diabetes duration and with increased blood glucose. There was a positive correlation between IGF-I and insulin levels and also an inverse correlation between IGF-I and IGFBP-1 levels. The IGFBP-1 levels were twofold higher than reported in non-diabetic individuals. In multiple stepwise correlation analysis, 37% of the variability in IGFBP-1 could be explained by BMI, IGF-I SD score, age, IGF-I, and fasting blood glucose. CONCLUSION: Our study indicates that influence of type 2 diabetes on IGF-I bioavailability in individual patients is modulated by insulin, body weight (presumably reflecting insulin sensitivity) and metabolic control. Furthermore, increased levels of IGFBP-1 are strongly associated with decreased b-cell function in type 2 diabetes mellitus. PMID- 9741825 TI - Neurotransmitters and their receptors in the islets of Langerhans of the pancreas: what messages do acetylcholine, glutamate, and GABA transmit? AB - Although neurotransmitters are present in pancreatic islets of Langerhans and can be shown to alter hormone secretion, their precise physiological roles in islet function and their cellular mechanisms of action are unclear. Recent research has identified specific neurotransmitter receptor isoforms in islets that may be important physiologically, because selective receptor agonists activate islet ion channels, modify intracellular [Ca2+], and affect secretion. This article focuses on the putative roles of acetylcholine, glutamate, and GABA in islet function. It has been hypothesized that acetylcholine potentiates insulin secretion by either promoting Ca release from cellular stores, activating a store depletion-activated channel, or activating a novel Na channel. GABA and glutamate, in contrast, have been proposed to mediate a novel paracrine signaling pathway whereby alpha- and beta-cells communicate within the islet. The evidence supporting these hypotheses will be critically evaluated. PMID- 9741826 TI - Estradiol and gonadotropin-releasing hormone (GnRH) interact to increase GnRH receptor expression in ovariectomized ewes after hypothalamic-pituitary disconnection. AB - Gonadotropin-releasing hormone (GnRH) receptor expression is regulated by estradiol and GnRH itself. The objective of this experiment was to determine the extent to which low levels of estradiol, similar to those observed during the transition from the luteal to the follicular phase of the estrous cycle, and GnRH interact to regulate expression of GnRH receptors and GnRH receptor mRNA. Ewes were ovariectomized (OVX) at least 2 wk prior to initiation of the experiment, and the pituitary gland was surgically disconnected from the hypothalamus to remove ovarian and hypothalamic inputs to the pituitary. Within 24 h after hypothalamic-pituitary disconnection, ewes received pulses of GnRH (250 ng/pulse) every 2 h for 6 d. At the end of 6 d, ewes were randomly assigned to treatments in a 2 x 2 factorial arrangement as follows: half of the animals received a single estradiol implant and half received an empty implant (placebo). At the same time, animals also received one of the following treatments: (1) saline or (2) GnRH (100 ng/pulse/2 h). Additionally, one group of ewes was ovariectomized, but not subjected to hypothalamic-pituitary disconnection (OVX controls). Blood samples were collected 15 min prior to each pulse of GnRH or saline and at 15-min intervals for 1 h after each pulse until tissues were collected and concentrations of luteinizing hormone (LH) were determined. Anterior pituitaries were collected 24 h after implant insertion to quantitate steady-state amounts of GnRH receptor mRNA and numbers of GnRH receptors. Mean LH was greatest in ovariectomized control ewes compared to all other treatments (p < 0.05). Mean LH and LH pulse amplitude in the placebo and GnRH-treated group most closely mimicked LH secretion in ovariectomized control animals. Mean LH and LH pulse amplitude were similar between both GnRH-treated groups (p < 0.05). Mean LH and LH pulse amplitude were significantly lower in all animals treated with saline compared to OVX controls (p < 0.05). Treatment with an estradiol implant and pulsatile GnRH increased (p < 0.05) relative amounts of GnRH receptor mRNA and the number of GnRH receptors compared to all other treatments. There were no differences in GnRH receptor expression between the remaining treatment groups (p > 0.05). Therefore, in OVX ewes after hypothalamic-pituitary disconnection, low levels of estradiol and GnRH are required to increase GnRH receptor mRNA and GnRH receptor numbers. Since we only observed an increase in GnRH receptor expression in the presence of both estradiol and GnRH, we conclude that there is a synergistic interaction between these two hormones in the regulation of GnRH receptor expression. PMID- 9741827 TI - Elevated proenkephalin-derived peptide levels in ACTH-producing adenomas: nucleus and cytoplasm localization. AB - The biosynthesis of met-enkephalin in human pituitary and human pituitary adenomas is still not well known. In this work, we studied the processing of proenkephalin-derived peptides in postmortem human pituitary (PMHP), ACTH producing adenomas (ACTH-PA), nonfunctioning adenomas (NFA), and GH-producing adenomas (GH-PA). ACTH-PA contained at least 10 times more proenkephalin-derived peptides than PMHP, NFA,and GH-PA. Proenkephalin processing was different in the four tested tissues. In ACTH-PA, proenkephalin was processed to high-, intermediate-, and low-mol-wt products. The highest met-enkephalin-containing peptides levels corresponded to intermediate and low-mol-wt materials, although met-enkephalinArg-Phe and synenkephalin immunoreactivity appeared only in high mol-wt peptides. In PMHP and NFA, met-enkephalin-Arg-Phe immunoreactivity was detected in intermediate- and low-mol-wt materials, and it was absent in GH-PA. Immunoblotting of ACTH-PA showed that met-enkephalin-Arg-Phe immunoreactivity corresponded to peptides of 44, 32-30, 27, and 17 kDa. The 32-30 and 17-kDa molecules were localized in the nuclear fraction where they were extracted after enzymatic digestion with DNase I. Plasmatic met-enkephalin levels did not increase in patients with Cushing's disease, suggesting that the pentapeptide stored in ACTH-PA was not released to the general circulation. In conclusion, we demonstrated that only ACTH-PA contained high levels of proenkephalin peptides, which were stored in cytoplasm organelles and in the nucleus, probably bound to chromatin. These results suggest an adenoma-specific physiological role of proenkephalin products. PMID- 9741828 TI - Effect of lipopolysaccharide on tumor necrosis factor and prolactin release from rat anterior pituitary cells. AB - TNF-alpha plays a critical role in the cascade of neuroendocrine events during inflammation and septic shock. It also affects the release of pituitary hormones and acts as a growth factor in immune and nonimmune cells. The aim of the present study was to investigate the release of TNF-alpha from rat anterior pituitary cells and the effect of the steroid medium on its release. Cultured anterior pituitary cells from lactating rats spontaneously released TNF-alpha. The presence of lipopolysaccharide (LPS, 0.1 microg/mL) in the culture medium significantly increased TNF-alpha release and inhibited prolactin release. Chronic estrogenization of ovariectomized rats or the presence of 17 beta estradiol in the culture medium also increased TNF-alpha release. LPS significantly stimulated TNF-alpha release in all groups and abrogated the estrogen-induced prolactin release. We also investigated the effect of TNF-alpha on prolactin release. The presence of TNF-alpha (50 ng/mL) in the culture medium inhibited prolactin release from anterior pituitary cells. These data show that anterior pituitary cells in culture release TNF-alpha and that this release is stimulated by estrogens. Our results also indicate that LPS inhibits prolactin release in an estrogenic environment, suggesting that TNF-alpha could affect pituitary hormone release during endotoxemia. PMID- 9741829 TI - Effect of repaglinide on insulin secretion in islets from rats infused for two days with a hypertonic solution of D-glucose. AB - This study investigates the insulin secretory responsiveness of pancreatic islets to repaglinide in an experimental model of B-cell glucose incompetence. Rats were infused for 2 d with a 1.67 M solution of D-glucose administered at a rate close to 2.8 mL/h. This resulted in a modest rise in glycemia, a severe increase in plasma insulin concentration, an increased sensitivity of B-cells to adrenergic stress, an abnormally high insulin output from isolated islets perifused in the presence of 16.7 mM D-glucose, and a paradoxical transient increase in insulin release from the islets in response to a fall in hexose concentration. The early increment in insulin output evoked by repaglinide, in the presence of 16.7 mM D glucose, was not lower in the islets from glucose-infused rats than in those from control rats. Moreover, when the meglitinide analog was administered concomitantly with the removal of D-glucose from the perifusion medium, the early response to repaglinide was further increased. Even after 24 min of glucose deprivation, the output of insulin by the islets from glucose-infused rats was higher in the presence of repaglinide than in its absence. These findings indicate that, in this model of B-cell dysfunction, the secretory responsiveness to repaglinide, as distinct from that to glucose, is fully preserved. Therefore, when taken into consideration together with prior observations, these findings argue in support of the use of this insulinotropic agent in the treatment of noninsulin-dependent diabetes. PMID- 9741831 TI - Regulation of prostaglandin F2alpha-receptor mRNA in human granulosa-luteal cells by human chorionic gonadotrophin and prostaglandin F2alpha. AB - This study examined the effects of prostaglandin F2alpha (PGF2alpha) and human chorionic gonadotropin (hCG) on the levels of PGF2alpha-receptor (PGF2alpha-R) mRNA and steroidogenesis, in the human granulosa luteal cell (hGLC). Human GLCs collected from patients undergoing in vitro fertilization, were cultured for 24 h, after which cells were exposed to culture media containing either vehicle, hCG (1IU/mL), or hCG plus PGF2alpha (10(-11)-10(-6) M), for a further 24 h. Following the treatment period, media were collected and stored (-20 degrees C) until assayed for progesterone and 17beta-estradiol (estradiol). Immediately following the treatment period, cells were extracted for total RNA. Transcripts for PGF2alpha-R were detected by PCR with two different sets of oligonucleotide primers based on the published human and rat PGF2alpha-R sequences. PCR products were confirmed to be those of PGF2alpha-R by size and by Southern blot hybridization with an internal oligo nucleotide probe. All experiments were performed a minimum of three times, on cells from a minimum of three separate patients. Prostaglandin F2alpha-R mRNA was significantly downregulated, whereas progesterone and estradiol production were significantly stimulated by hCG. Conversely, both low (10(-11)M) and high concentrations (10(-6) M) of PGF2alpha restored PGF2alpha-R mRNA levels to those of the controls, whereas steroidogenesis was significantly inhibited by these conditions. At a concentration of 10(-9)M PGF2alpha-R mRNA was barely detectable. Progesterone and estradiol production were inversely related to PGF2alpha-R levels, since hCG stimulated progesterone and estradiol production were completely restored in the presence of 10(-9) M PGF2alpha. Messenger RNA levels for the housekeeping gene beta-actin were unaltered by the above treatments. In conclusion, in the human granulosa luteal cell, PGF2alpha-R mRNA levels are inversely related to hCG stimulated steroidogenesis (which was biphasic in nature). Moreover, in the presence of hCG, PGF2alpha downregulates its receptor mRNA, thus providing a potential form of negative feedback on its own actions, which may be important in rescuing the corpus luteum from PGF2alpha-mediated luteolysis should pregnancy occur. PMID- 9741830 TI - Expression of ovine insulin-like growth factor-1 (IGF-1) stimulates alveolar bud development in mammary glands of transgenic mice. AB - To determine whether murine mammary growth is modulated by local insulin-like growth factor-1 (IGF-1) production, expression of recombinant IGF-1 was directed to the mammary glands of transgenic mice using an ovine prepro IGF-1 cDNA under control of the mouse mammary tumor virus-long terminal repeat (MMTV-LTR) promoter. Bioactivity of recombinant IGF-1 in transgenic mouse milk extracts was demonstrated by a concentration-dependent increase in [3H]thymidine incorporation in clonal bovine mammary epithelial cells (MAC-T) compared with control mouse milk extracts; moreover, addition of excess recombinant human insulin-like growth factor binding protein-3 (rhlGFBP-3) abolished the increase in [3H]thymidine incorporation attributed to recombinant IGF-1 in transgenic mouse milk. Recombinant IGF-1 was produced in mammary tissue of virgin and pregnant transgenic mice, and secreted into milk of lactating mice. However, recombinant IGF-1 was not detected in serum from transgenic mice; and ligand blot analysis of serum insulin-like growth factor binding proteins (IGFBPs) indicated no differences owing to transgene presence. In peripubertal virgin mice at 49 d of age, the frequency of appearance of mammary alveolar buds was significantly higher in MMTV-IGF-1 than in CD-1 mice, and was unaffected by ovariectomy or estradiol treatment. In conclusion, mammary synthesis of recombinant IGF-1 enhances the rate of development of alveolar buds in mammary glands of virgin transgenic mice. PMID- 9741832 TI - Gap junctional proteins, connexin 26, 32, and 43 in sheep ovaries throughout the estrous cycle. AB - Ovarian follicles from days 13, 14, 15, and 16 and corpora lutea (CL) from days 2, 4, 8, 12, and 15 of the estrous cycle were evaluated for the presence of connexins by immunohistochemistry. In addition, CL from days 5, 10, and 15 of the estrous cycle were used for immunofluorescent detection of Cx43 followed by image analysis, and for Western immunoblot. In all tissues, staining for all connexins appeared punctate, indicating the presence of assembled gap junctions. Cx26 was present in the ovarian surface epithelium, stroma, and blood vessels within the stroma and hilus, and in the CL. In healthy antral follicles, Cx26 was present only in the theca layer, whereas Cx43 was present in granulosa and theca layers. In the majority of atretic follicles, connexins were not detected, but in 13% of the atretic follicles, Cx43 was present in the theca layer. Cx32 was detected in the blood vessels of ovarian stroma and in the CL, and Cx43 was detected in the CL. Localization and/or expression of connexins depended on stage of luteal development. Western analysis demonstrated that expression of Cx32 in luteal tissues was similar across the estrous cycle. The area of positive staining for Cx43 and expression of Cx43 in luteal tissues decreased (p < 0.05) as the estrous cycle progressed. The pattern of expression of connexins indicates that gap junctional proteins may be important in the regulation of folliculogenesis and follicular atresia, as well as growth, differentiation, and regression of the CL. PMID- 9741833 TI - A distal regulatory region of the insulin-like growth factor binding protein-2 (IGFBP-2) gene interacts with the basic helix-loop-helix transcription factor, AP 4. AB - Insulin-like growth factor binding protein-2 (IGFBP-2), the predominant IGFBP in the fetal circulation and an induced protein during several types of malignancies, belongs to a family of structurally related proteins that bind the mitogens, IGF-1 and IGF-2. The present study focused on functional analysis of the 5 '-flanking region (approximately 1.3 kb) of the IGFBP-2 gene to identify nuclear factors that mediate hepatic transcription of this gene. Luciferase (LUC) reporter constructs containing progressive deletions of 5'-flanking DNA and the intact promoter of the porcine IGFBP-2 gene were examined for functional activity by transient transfection of human HepG2 liver cells. LUC activity of the transfected reporter gene driven by the IGFBP-2 promoter and flanking sequences to -1397 (numbering relative to initiation codon at +1) was 22-fold higher than that of promoterless parent LUC vector. This activity was decreased by 60% with deletion of sequences to -874 bp, and dropped to basal levels with further truncation to -764 bp. The region between -874 and -765 bp (110 bp) functioned as a potent stimulator of heterologous SV40 promoter activity (110 bp/SV40-LUC construct) and was found to contain two noncontiguous basic helix-loop-helix (bHLH) transcription factor binding motifs (E-boxes [CAN NTG]: CACCTG and CAAATG). In electrophoretic mobility shift assays, nuclear proteins prepared from HepG2 cells formed two complexes (C1, C2) with double-stranded oligonucleotides containing either HLH sequence, mutations of which resulted in loss of complex formation. Southwestern blot analysis identified an HepG2 nuclear protein with molecular mass of 48 kDa, similar to that of the bHLH transcription factor AP-4, which bound the CACCTG motif. Cotransfection of HepG2 cells with the 110-bp/SV40 LUC construct and an expression vector encoding human AP-4 increased IGFBP-2 fragment-dependent SV40 promoter activity by 16-fold. This AP-4-mediated stimulation was lost following block mutation of both bHLH motifs within the IGFBP-2 110-bp fragment. Results demonstrate the functional importance of sequences upstream of the promoter in IGFBP-2 gene transcription and identify a novel mechanism by which bHLH proteins potentially may affect cell proliferation and differentiation via induction of IGFBP-2 synthesis. PMID- 9741834 TI - Effect of transforming growth factor-beta1 on parathyroid hormone-related protein secretion and mRNA expression by normal human keratinocytes in vitro. AB - Parathyroid hormone-related protein (PTHrP) is produced by a wide range of neoplastic and normal cells, including keratinocytes where it may regulate growth and differentiation. Transforming growth factor-beta (TGF-beta) is a growth factor produced by many cells, including keratinocytes where it regulates epidermal homeostasis. TGF-beta has been reported to be cosecreted with PTHrP in some neoplasms and to stimulate PTHrP production by neoplastic keratinocytes. However, the effects of TGF-beta on PTHrP production by normal keratinocytes are not well characterized. In this study, we investigated the effects of endogenous and exogenous TGF-beta on PTHrP production by normal human foreskin keratinocytes. PTHrP secretion, mRNA expression, and mRNA transcription in vitro were determined by N-terminal radioimmunoassay, ribonuclease protection assay, and transient transfections. PTHrP production and secretion of latent TGF-beta activity were greatest in proliferating keratinocytes prior to and at confluence of monolayer cultures. TGF-beta1 increased PTHrP mRNA expression by normal keratinocytes in a dose-dependent manner with maximal stimulation at 6-1 2 h after treatment. In addition, keratinocytes treated with a monoclonal anti-TGF beta antibody expressed decreased levels of PTHrP mRNA. The increased levels of PTHrP mRNA following TGF-beta1 treatment were owing, at least partly, to an increase in PTHrP mRNA stability. TGF-beta1 failed to activate transcription of the luciferase reporter gene driven by either the human or mouse PTHrP promoters. In conclusion, TGF-beta1 functions as a paracrine or autocrine regulator of PTHrP production in normal human keratinocytes, and this may play a role in the regulation of keratinocyte proliferation or differentiation. PMID- 9741835 TI - Stepwise activation of the gonadotropic signal transduction pathway, and the ability of prostaglandin F2alpha to inhibit this activated pathway. AB - Through selective activation of the gonadotropic signal transduction pathway, we have determined the probable site of the antigonadotropic effects of prostaglandin F2alpha (PGF2alpha) in the human granulosa-luteal cell (hGLC). The gonadotropic signal transduction pathway was activated at the level of the receptor (luteinizing hormone and beta-adrenergic), stimulatory G protein (Gs), adenylate cyclase (AC), and protein kinase A (PKA) by human chorionic gonadotropin (hCG) and isoproterenol (Iso), cholera toxin (CTX), forskolin, and dibutryl cAMP (Db cAMP), respectively. Concomitantly, the ability of PGF2alpha to inhibit progesterone production in response to the activation of this cascade at these different levels was examined. hGLCs were obtained from in vitro fertilization patients and were precultured for 8 d in Medium 199 supplemented with fetal bovine serum (M199; 10% FBS). Following the preculture period, cells were treated with either vehicle or one of the above activators of the gonadotropic pathway, either in the absence or presence of PGF2alpha (in M199; No FBS). Following the treatment period, media were collected and assayed for progesterone by RIA. Prostaglandin F2alpha (10(-6) M) significantly inhibited hCG (1 IU/mL), Iso (10(-5) M), CTX (1 microg/mL), and forskolin- (10(-5) M) stimulated progesterone production. Conversely, PGF2alpha did not inhibit progesterone production stimulated by a saturating concentration of Db cAMP (10( 6) M). The ability of PGF2alpha to inhibit hCG- or CTX-stimulated progesterone production was attenuated by pertussis toxin (PTX; 50 ng/mL). In conclusion, through a pertussis toxin-sensitive G protein, PGF2alpha inhibits progesterone production at a level below AC, and above the activation of PKA by cAMP. PMID- 9741837 TI - The cardiovascular effects of porcine relaxin in Brattleboro rats. AB - The effects of porcine relaxin on arterial blood pressure, heart rate, and the release of vasopressin and oxytocin were investigated in homozygous diabetes insipidus (di/di) Brattleboro and Long-Evans rats. Acute iv injection of relaxin (5 microg) caused a significant increase in mean arterial, systolic and diastolic blood pressures in Long-Evans rats compared with control injections of saline, but had no pressor effect in Brattleboro rats. Circulating concentrations of vasopressin were also significantly elevated above baseline in the Long-Evans rats 1 min after relaxin treatment, but remained undetectable in the relaxin treated Brattleboro rats. Relaxin increased heart rate in both groups of animals 4 min after injection. The chronotropic effect of relaxin was, however, attenuated in the Brattleboro rats. Intravenous relaxin injection also caused a significant increase in plasma oxytocin concentrations 5 min posttreatment in both the Long-Evans and Brattleboro rats. The change in plasma oxytocin above basal concentrations was significantly greater in Brattleboro rats compared with Long-Evans controls. The data in this study demonstrate that iv relaxin increases heart rate, but not arterial blood pressure in Brattleboro rats. Furthermore, the relaxin-induced release of oxytocin in Brattleboro rats does not result in an acute pressor response. PMID- 9741836 TI - Stress promotes development of ovarian cysts in rats: the possible role of sympathetic nerve activation. AB - Activation of the sympathetic innervation precedes the induction of polycystic ovaries in rats given estradiol valerate (EV). The mechanism of induction by EV may thus involve both direct and neurogenic components. We tested this hypothesis using a combined cold and restraint stress to induce an increase in sympathetic tone, including that of the ovarian sympathetic nerves. Three weeks after the start of stress we found: 1. An increase in the content of norepinephrine (NE) in the celiac ganglion. 2. An increase in the release of NE from the ovary. 3. An unchanged NE uptake by the ovary. 4. An unchanged content of NE in the ovary. The ovarian content of neuropeptide Y (NPY) (colocalized with NE) was significantly decreased. These results suggest that NE synthesis and its secretion are increased during this period and correlate with the increase in secretion of androgens and estradiol, the development of precystic follicles, and a decrease in the ovulatory rate. After 11 wk, NE release had returned to control values, whereas the ovarian NE content had risen significantly, suggesting a maintained high rate of NE synthesis. In the ovary, NPY contents, steroid secretion, morphology, and ovulation had returned to the control state. These results suggest the participation of an extraovarian factor that might act locally to control the release of NE from the ovary, and further support the hypothesis that increased sympathetic activity plays a role in the development and maintenance of ovarian cysts. PMID- 9741839 TI - Plasma disappearance of exogenous erythropoietin in mice under different experimental conditions. AB - Erythropoietin (EPO) is a glycoprotein hormone produced primarily in the kidneys and to a lesser extent in the liver that regulates red cell production. Most of the studies conducted in experimental animals to assess the role of EPO in the regulation of erythropoiesis were performed in mouse models. However, little is known about the in vivo metabolism of the hormone in this species. The present study was thus undertaken to measure the plasma tl/2 of radiolabeled recombinant human EPO (rh-EPO) in normal mice as well as in mice with altered erythrocyte production rates (EPR), plasma EPO (pEPO) titer, marrow responsiveness, red cell volume, or liver function. Adult CF-1 mice of both sexes were used throughout. For the EPO life-span studies, 30 mice in each experiment were intravenously injected with 600,000 cpm of 125l-rh-EPO and bled by cardiac puncture in groups of five every hour for 6 h. Trichloroacetic acid (TCA) was added to each plasma sample and the radioactivity in the precipitate measured in a gamma-counter. EPO, pEPO, marrow responsiveness, or red cell volume were altered by either injections of rh-EPO, 5-fluorouracil, or phenylhydrazine, or by bleeding, or red cell transfusion. Liver function was altered by CI4C administration. In the normal groups of mice, the estimated tl/2 was 182.75+/-14.4 (SEM) min. The estimated tl/2 of the other experimental groups was not significantly different from normal. These results, therefore, strongly suggest that the clearance rate of EPO in mice is not subjected to physiologic regulation and that pEPO titer can be really taken as the reflection of the EPO production rate, at least in the experimental conditions reported here. PMID- 9741838 TI - GLP-1 receptors in golden Syrian hamster islets: identification and functional characterization. AB - This study aims at the identification and functional characterization of glucagon like peptide 1 (7-36) amide (GLP-1) receptor in islets from Golden Syrian hamsters. Using a polyclonal antibody against rat GLP-1 receptors, Western blotting of the islet proteins revealed two major bands of 44 and 70 kDa, similar to those found in rat islets, RINm5F cells, and HIT-T15 cells. In Northern blots, transcripts of 2.7, 3.6 and 3.7 kb were observed in rat islets and RINm5F cells after hybridization with rat GLP-1 receptor cDNA probes of either 21 9 bp or 1.5 kb. Such was not the case in either hamster islets or HIT-T15 cells. However, a single 3.6-kb transcript was observed in the latter two cases when a human GLP-1 receptor cDNA probe of 1.6 kb was used for hybridization. In the isolated perfused pancreas of Golden Syrian hamsters, a rise in D-glucose concentration from 3.3 to 8.3 mM caused a biphasic stimulation of insulin release, which was further increased by either GLP-1 or glucagon (10(-9)M each). The enhancing action of GLP-1 on glucose-stimulated insulin secretion was much more marked than that of glucagon. The rise in D-glucose concentration decreased by 46+/-4% the release of glucagon, but GLP-1 failed to exert any obvious effect on glucagon secretion in the presence of 8.3 mM D-glucose. These results indicate that GLP-1 receptors are expressed in islets of Golden Syrian hamsters with an extracellular part possessing the same immunoreactivity as the rat islet GLP-1 receptors. The expression of the mRNA for the GLP-1 receptor differs, however, from that found in rat or human islets. PMID- 9741840 TI - Analytical shape computation of macromolecules: I. Molecular area and volume through alpha shape. AB - The size and shape of macromolecules such as proteins and nucleic acids play an important role in their functions. Prior efforts to quantify these properties have been based on various discretization or tessellation procedures involving analytical or numerical computations. In this article, we present an analytically exact method for computing the metric properties of macromolecules based on the alpha shape theory. This method uses the duality between alpha complex and the weighted Voronoi decomposition of a molecule. We describe the intuitive ideas and concepts behind the alpha shape theory and the algorithm for computing areas and volumes of macromolecules. We apply our method to compute areas and volumes of a number of protein systems. We also discuss several difficulties commonly encountered in molecular shape computations and outline methods to overcome these problems. PMID- 9741841 TI - Analytical shape computation of macromolecules: II. Inaccessible cavities in proteins. AB - The structures of proteins are well-packed, yet they contain numerous cavities which play key roles in accommodating small molecules, or enabling conformational changes. From high-resolution structures it is possible to identify these cavities. We have developed a precise algorithm based on alpha shapes for measuring space-filling-based molecular models (such as van der Waals, solvent accessible, and molecular surface descriptions). We applied this method for accurate computation of the surface area and volume of cavities in several proteins. In addition, all of the atoms/residues lining the cavities are identified. We use this method to study the structure and the stability of proteins, as well as to locate cavities that could contain structural water molecules in the proton transport pathway in the membrane protein bacteriorhodopsin. PMID- 9741842 TI - Crystal structure of a complex formed between proteolytically-generated lactoferrin fragment and proteinase K. AB - Lactoferrin is an iron binding glycoprotein with a molecular weight of 80 kDa. The molecule is divided into two lobes representing the N-terminal and C-terminal halves of the polypeptide chain, each containing an iron binding site. The serine proteinases such as trypsin, chymotrypsin, and pepsin hydrolyze lactoferrin into two unequal halves while proteinase K divides this protein into two equal halves. In the first step of hydrolysis by proteinase K, the C- and N-lobes, each having a molecular weight of approximately 40 kDa, are generated. In the next step, the lobes are further hydrolyzed into small molecular weight peptides. The proteinase K isolated from the hydrolyzed product does not show enzymatic activity suggesting that the enzyme is inhibited. Furthermore, the hydrolysis experiments on N-lobe and C-lobe showed that the inhibitory fragment came from the C-lobe. The purified lactoferrin fragment was found to be a decapeptide with an amino acid sequence of H2N-Val-Ala-Gln-Gly-Ala-Ala-Gly-Leu-Ala-COOH. The complex formed between proteinase K and lactoferrin fragment was crystallized by microdialysis. The crystals belonged to the monoclinic space group P2(1) with cell dimensions a = 44.4 A, b = 38.6 A, c = 79.2 A, beta = 105.8 degrees and Z = 2. The crystal structure has been determined at 2.4 A resolution. It has been refined to an R factor of 0.163 for 9044 reflections. The Lf-fragment forms several intermolecular interactions with proteinase K. The Ser-224 Ogamma and His-57 N epsilon2 move away to a distance of 3.68 A in the complex. In the crystal structure, Gln-3I (I indicates inhibitor i.e., lactoferrin fragment) is involved in a direct intermolecular interaction with a symmetry related proteinase K molecule through a strong hydrogen bond with Asp-254. The mode of intermolecular interactions in the complex conformational features of the enzyme and placement of the fragment with respect to the enzyme resemble with the molecular complex of proteinase K with its natural inhibitor PKI3 from wheat. PMID- 9741843 TI - Calculation of HyHel10-lysozyme binding free energy changes: effect of ten point mutations. AB - The change in free energy of binding of hen egg white lysozyme (HEL) to the antibody HyHel-10 arising from ten point mutations in HEL (D101K, D101G, K96M, K97D, K97G, K97G, R21E, R21K, W62Y, and W63Y) was calculated using a combination of the finite difference Poisson-Boltzmann method for the electrostatic contribution, a solvent accessible surface area term for the non-polar contribution, and rotamer counting for the sidechain entropy contribution. Comparison of experimental and calculated results indicate that because of pKa shifts in some of the mutated residues, primarily those involving Aspartate or Glutamate, proton uptake or release occurs in binding. When this effect was incorporated into the binding free energy calculations, the agreement with experiment improved significantly, and resulted in a mean error of about 1.9 kcal/mole. Thus these calculations predict that there should be a significant pH dependence to the change in binding caused by these mutations. The other major contributions to binding energy changes comes from solvation and charge charge interactions, which tend to oppose each other. Smaller contributions come from nonpolar interactions and sidechain entropy changes. The structures of the HyHel 10-HEL complexes with mutant HEL were obtained by modeling, and the effect of the modeled structure on the calculations was also examined. "Knowledge based" modeling and automatic generation of models using molecular mechanics produced comparable results. PMID- 9741844 TI - Biphasic denaturation of human placental alkaline phosphatase in guanidinium chloride. AB - Human placental alkaline phosphatase is a membrane-anchored dimeric protein. Unfolding of the enzyme by guanidinium chloride (GdmCl) caused a decrease of the fluorescence intensity and a large red-shifting of the protein fluorescence maximum wavelength from 332 to 346 nm. The fluorescence changes were completely reversible upon dilution. GdmCl induced a clear biphasic fluorescence spectrum change, suggesting that a three-state unfolding mechanism with an intermediate state was involved in the denaturation process. The half unfolding GdmCl concentrations, [GdmCl]0.5, corresponding to the two phases were 1.45 M and 2.50 M, respectively. NaCl did not cause the same effect as GdmCl, indicating that the GdmCl-induced biphasic denaturation is not a salt effect. The decrease in fluorescence intensity was monophasic, corresponding to the first phase of the denaturation process with [GdmCl]0.5 = 1.37 M and reached a minimum at 1.5 M GdmCl, where the enzyme remained completely active. The enzymatic activity lost started at 2.0 M GdmCl and was monophasic but coincided with the second-phase denaturation with [GdmCl]0.5 = 2.46 M. Inorganic phosphate provides substantial protection of the enzyme against GdmCl inactivation. Determining the molecular weight by sucrose-density gradient ultracentrifugation revealed that the enzyme gradually dissociates in both phases. Complete dissociation occurred at [GdmCl] > 3 M. The dissociated monomers reassociated to dimers after dilution of the GdmCl concentration. Refolding kinetics for the first-phase denaturation is first-order but not second-order. The biphasic phenomenon thereby was a mixed dissociation denaturation process. A completely folded monomer never existed during the GdmCl denaturation. The biphasic denaturation curve thereby clearly demonstrates an enzymatically fully active intermediate state, which could represent an active site structure intact and other structure domains partially melted intermediate state. PMID- 9741845 TI - A new method for predicting binding free energy between receptor and ligand. AB - A practical method to estimate binding free energy, deltaG(bind), of a given ligand structure to the target receptor has been developed. The method assumes that deltaG(bind) is given by the summation of intermolecular interaction energy, deltaG(inter), and partial desolvation energy, deltaG(desolv). DeltaG(desolv) is calculated from the buried surface area in the complex between the ligand and receptor, based on solvation energy, deltaG(solv), formulated by an equation which can be calibrated with observed values. Then, the method was applied to arabinose-binding protein (ABP) and dihydrofolate reductase (DHFR), after recalibrating the weights for deltaG(inter) and each term of deltaG(desolv) using observed deltaG(bind) data for 29 known ligands to avidin (AV). The usefulness of our method was confirmed by the fact that correlation coefficients between the calculated and observed deltaG(bind)'s in AV, ABP and DHFR were 0.92, 0.77, and 0.88, whereas the corresponding values obtained by simple force field calculation were 0.79, 0.30, and 0.79, respectively. Further investigations to improve the method and validate the parameters are in progress. PMID- 9741846 TI - Screening a peptidyl database for potential ligands to proteins with side-chain flexibility. AB - The three key challenges addressed in our development of SPECITOPE, a tool for screening large structural databases for potential ligands to a protein, are to eliminate infeasible candidates early in the search, incorporate ligand and protein side-chain flexibility upon docking, and provide an appropriate rank for potential new ligands. The protein ligand-binding site is modeled by a shell of surface atoms and by hydrogen-bonding template points for the ligand to match, conferring specificity to the interaction. SPECITOPE combinatorially matches all hydrogen-bond donors and acceptors of the screened molecules to the template points. By eliminating molecules that cannot match distance or hydrogen-bond constraints, the transformation of potential docking candidates into the ligand binding site and the shape and hydrophobic complementarity evaluations are only required for a small subset of the database. SPECITOPE screens 140,000 peptide fragments in about an hour and has identified and docked known inhibitors and potential new ligands to the free structures of four distinct targets: a serine protease, a DNA repair enzyme, an aspartic proteinase, and a glycosyltransferase. For all four, protein side-chain rotations were critical for successful docking, emphasizing the importance of inducible complementarity for accurately modeling ligand interactions. SPECITOPE has a range of potential applications for understanding and engineering protein recognition, from inhibitor and linker design to protein docking and macromolecular assembly. PMID- 9741847 TI - Dictionary of recurrent domains in protein structures. AB - The rapid growth in the number of experimentally determined three-dimensional protein structures has sharpened the need for comprehensive and up-to-date surveys of known structures. Classic work on protein structure classification has made it clear that a structural survey is best carried out at the level of domains, i.e., substructures that recur in evolution as functional units in different protein contexts. We present a method for automated domain identification from protein structure atomic coordinates based on quantitative measures of compactness and, as the new element, recurrence. Compactness criteria are used to recursively divide a protein into a series of successively smaller and smaller substructures. Recurrence criteria are used to select an optimal size level of these substructures, so that many of the chosen substructures are common to different proteins at a high level of statistical significance. The joint application of these criteria automatically yields consistent domain definitions between remote homologs, a result difficult to achieve using compactness criteria alone. The method is applied to a representative set of 1,137 sequence-unique protein families covering 6,500 known structures. Clustering of the resulting set of domains (substructures) yields 594 distinct fold classes (types of substructures). The Dali Domain Dictionary (http://www.embl-ebi.ac.uk/dali/) not only provides a global structural classification, but also a comprehensive description of families of protein sequences grouped around representative proteins of known structure. The classification will be continuously updated and can serve as a basis for improving our understanding of protein evolution and function and for evolving optimal strategies to complete the map of all natural protein structures. PMID- 9741848 TI - Crystal structures of HLA-A*0201 complexed with antigenic peptides with either the amino- or carboxyl-terminal group substituted by a methyl group. AB - The crystal structures of class I major histocompatibility complex (MHC) molecules complexed with antigenic peptides revealed a network of hydrogen bonds between the charged amino- and carboxyl-termini of the peptides and conserved MHC residues at both ends of the peptide binding site. These interactions were shown to contribute substantially to the stability of class I MHC/peptide complexes by thermal denaturation studies using synthetic peptides in which either the amino- or carboxyl-terminal group is substituted by a methyl group. Here we report crystal structures of HLA-A*0201 complexed with these terminally modified synthetic peptides showing that they adopt the same bound conformation as antigenic peptides. A number of variations in peptide conformation were observed for the terminally modified peptides, including in one case, a large conformational difference in four central peptide residues that is apparently caused by the lattice contact. This is reminiscent of the way binding a T-cell receptor changed the conformation of central residues of an MHC-bound peptide. The structures determined identify which conserved hydrogen bonds are eliminated in terminally substituted peptides and suggest an increased energetic importance of the interactions at the peptide termini for MHC-peptide stability. PMID- 9741849 TI - Folding mechanism of three structurally similar beta-sheet proteins. AB - The folding mechanism of cellular retinoic acid binding protein I (CRABP I), cellular retinol binding protein II (CRBP II), and intestinal fatty acid binding protein (IFABP) were investigated to determine if proteins with similar native structures have similar folding mechanisms. These mostly beta-sheet proteins have very similar structures, despite having as little as 33% sequence similarity. The reversible urea denaturation of these proteins was characterized at equilibrium by circular dichroism and fluorescence. The data were best fit by a two-state model for each of these proteins, suggesting that no significant population of folding intermediates were present at equilibrium. The native states were of similar stability with free energies (linearly extrapolated to 0 M urea, deltaGH2O) of 6.5, 8.3, and 5.5 kcal/mole for CRABP I, CRBP II, and IFABP, respectively. The kinetics of the folding and unfolding processes for these proteins was monitored by stopped-flow CD and fluorescence. Intermediates were observed during both the folding and unfolding of all of these proteins. However, the overall rates of folding and unfolding differed by nearly three orders of magnitude. Further, the spectroscopic properties of the intermediate states were different for each protein, suggesting that different amounts of secondary and/or tertiary structure were associated with each intermediate state for each protein. These data show that the folding path for proteins in the same structural family can be quite different, and provide evidence for different folding landscapes for these sequences. PMID- 9741850 TI - Molecular dynamics simulations of human carbonic anhydrase II: insight into experimental results and the role of solvation. AB - In this paper, the carbonic anhydrase II (CA II) enzyme active site is modeled using ab initio calculations and molecular dynamics simulations to examine a number of important issues for the enzyme function. It is found that the Zn2+ ion is dominantly tetrahedrally coordinated, which agrees with X-ray crystallographic studies. However, a transient five-fold coordination with an extra water molecule is also found. Studies of His64 conformations upon a change in the protonation states of the Zn-bound water and the His64 residue also confirm the results of an X-ray study which suggest that the His64 conformation is quite flexible. However, the degree of water solvation is found to affect this behavior. Water bridge formation between the Zn-bound water and the His64 residue was found to involve a free energy barrier of 2-3 kcal/mol and an average lifetime of several picoseconds, which supports the concept of a proton transfer mechanism through such a bridge. Mutations of various residues around the active site provide further insight into the corresponding experimental results and, in fact, suggest an important role for the solvent water molecules in the CA II catalytic mechanism. PMID- 9741851 TI - Folding the ribonuclease H domain of Moloney murine leukemia virus reverse transcriptase requires metal binding or a short N-terminal extension. AB - Reverse transcriptase (RT) is a modular enzyme carrying polymerase and ribonuclease H (RNase H) activities in separable domains. Retroviral replication requires both of these activities. The RNase H domain is responsible for hydrolysis of the RNA portion of RNA x DNA hybrids, and this activity requires the presence of divalent cations (Mg2+ or Mn2+) that bind its active site. This domain is a part of a large family of homologous RNase H enzymes of which the RNase HI protein from Escherichia coli is the best characterized. Although the isolated RNase H domain from human immunodeficiency virus RT is inactive, the Moloney murine leukemia virus (MMLV) domain is active in the absence of the polymerase domain, making functional studies more accessible. Using circular dichroism spectroscopy, we characterized the stability and folding of two different fragments of MMLV RT that retain RNase H activity. The smaller fragment corresponding to the 157 C-terminal residues of RT is predominantly unfolded in the absence of divalent cations, but folding can be induced by the addition of metal. The larger fragment corresponding to the 175 C-terminal residues, however, is stably folded in the absence of metal. Thus, an 18 residue N-terminal extension outside the region homologous to E. coli RNase HI is important for the structural stability of the RNase H domain of MMLV RT. Therefore, this region should be considered part of the RNase H domain. PMID- 9741852 TI - Some new multiple-test procedures for dose finding. AB - In Tamhane, Hochberg, and Dunnett (1) we focused primarily on step-down test procedures based on contrasts among the sample means to find the minimum effective dose in a dose-response study. In the present article we use the global tests of Bartholomew (2,3) and Hayter (4) in these step-down procedures. We also propose a new step-down procedure that permits tests based on a class of contrasts [step and basin contrasts of Ruberg (5) are examples of such contrasts] that could not be used with the step-down procedures studied in our previous paper because of lack of control of the familywise error rate. A simulation study to compare the four procedures proposed in the present paper with the top four procedures from the previous article is carried out. It is found that the step down procedure based on Bartholomew's test and the new step-down procedure based on step and modified basin contrasts generally perform better than the other procedures for a wide range of dose-response profiles. PMID- 9741853 TI - Sample size in clinical trials with dichotomous endpoints: use of covariables. AB - In many clinical trials, the primary endpoint is dichotomous. In this article, we examine the possibility of reducing the required sample size by removing variation associated with baseline covariables. Three measures are used to study the size of the reduction. Simulation studies based on a database of head trauma and of stroke patients suggested that a substantial reduction in the sample size can be achieved when the correlation between the endpoint and covariables is strong. A simple ad hoc formula for approximating the required sample size is proposed. PMID- 9741854 TI - Joint equivalence of means and variances of two populations. AB - Clinical equivalence has almost exclusively been treated from the view of average equivalence. However, as in bioequivalence, there has been recent interest in more general definitions of clinical equivalence. In particular, Bauer and Bauer (1) have investigated the possibility of the use of a pair of tests for the equality of means and variances taking account of the multiplicity issues involved. In bioequivalence, a number of authors have considered a Bayesian approach; in this article we generalize these previous Bayesian approaches to this more general problem. We investigate the operating characteristics of a decision procedure based on the posterior probability that the parameters lie within a prespecified region of equivalence. PMID- 9741855 TI - Sensitivity of parametric link functions in generalized linear models. AB - A common method of choosing the link function in generalized linear models is to specify a parametric link family indexed by unknown parameters. The maximum likelihood estimates of such link parameters, however, may often depend on one or several extreme observations. Diagnostics are derived to assess the sensitivity of the parametric link analysis. Two examples demonstrate that the proposed diagnostics can identify jointly influential observations on the link even when masking is present. PMID- 9741856 TI - Optimal selection procedures for abbreviated area under the curve (AAUC) of blood concentration versus time for drug blood concentration levels. AB - It has become common to collect blood samples from patients for phase II/III clinical trials to investigate pharmacokinetic/pharmacodynamic relationships. However, the frequency of blood samples drawn from patients is limited due to clinical or pharmacoeconomic reasons. We discuss some sampling schemes for an immunosuppressive drug in phase III studies. Selection criteria and statistical approaches are discussed to select blood-sampling time points in four scenarios from a single study and from combined multiple studies: (1) Area under the curve (AUC) only without constraint, (2) AUC and Cmax simultaneously without constraint, (3) AUC only with constraint(s), and (4) AUC and Cmax simultaneously with constraint(s). PMID- 9741857 TI - Regression spline models and model calibration in the identification of protein storage conditions. AB - The study of protein-construct activity provides valuable insight in the early stages of drug discovery. Identification of optimal storage conditions that maintain activity of the constructs is an important issue. In the study reported herein, a space-filling design was used to assess the effects of eight design variables--buffer, pH, NaCl, protein concentration, reducing agent, detergent, MgCl2, and temperature--on protein activity. A regression spline analysis is presented, and settings of the explanatory factors resulting in the best predicted protein activity over the explored space are identified. The models are selected initially based on the Akaike information criterion and later assessed via root mean square error and cross-validation root mean square error. Detergent, buffer, temperature, and smooth functions of pH and protein concentration were found to have large effects on protein activity. The models were calibrated via calculation of the empirical distribution of the cross validation root mean square error. In this framework several models provide a similar fit, and further experimentation is required for more definitive conclusions regarding protein storage conditions to be made. The cross-validation root mean square error calibration of models is recommended for applications involving comparisons of models with respect to their predictive ability. PMID- 9741858 TI - Protein construct storage: Bayesian variable selection and prediction with mixtures. AB - Determining optimal conditions for protein storage while maintaining a high level of protein activity is an important question in pharmaceutical research. A designed experiment based on a space-filling design was conducted to understand the effects of factors affecting protein storage and to establish optimal storage conditions. Different model-selection strategies to identify important factors may lead to very different answers about optimal conditions. Uncertainty about which factors are important, or model uncertainty, can be a critical issue in decision-making. We use Bayesian variable selection methods for linear models to identify important variables in the protein storage data, while accounting for model uncertainty. We also use the Bayesian framework to build predictions based on a large family of models, rather than an individual model, and to evaluate the probability that certain candidate storage conditions are optimal. PMID- 9741859 TI - Bayesian decision procedures based on logistic regression models for dose-finding studies. AB - Early-phase clinical trials, conducted to determine the appropriate dose of an experimental drug to take forward to later trials, are considered. The objective is to find the dose associated with some low probability of an adverse event. A Bayesian model is presented, and a decision-theoretic procedure for finding the optimal doses for each of a series of cohorts of subjects is derived. The procedure is flexible and can easily be conducted using standard statistical software. The results of simulations investigating the properties of the procedure are presented. PMID- 9741860 TI - Approximately optimal designs for phase II clinical studies. AB - There is no consensus on determination of sample size in phase II clinical trials. The use of Bayesian decision theory has been proposed by Stallard (1), among others. In this article, optimal three-stage designs are obtained using decision theory. These are compared with procedures proposed by Schoenfeld (2), Ensign et al. (3), and Chen et al. (4) and the sequential probability ratio test of Wald (5) and Barnard (6). The three-stage procedures are shown to be close to the true optimal test; the sequential probability ratio test is easier to obtain and only marginally inferior. Because optimality of the decision-theory approach depends on accurate specification of costs and profits, an assessment is made of the sensitivity of the proposed procedures to a range of gain function parameter values. PMID- 9741861 TI - Sample size determination for controlling the upper confidence limit of incidence rate of a binomial endpoint. AB - Assume that in a comparative clinical study the primary endpoint is a binary event, such as life or death. A new treatment or therapy is tested for a significant reduction of the incidence of the binary event compared with a control group. Another objective is to ensure that the incidence in the new treatment group is below some clinically acceptable value. This is done by calculating the exact upper 95% confidence limit for the probability of the event. The study is considered successful if the upper confidence limit is lower than a historical threshold, as well as if there is a significant reduction in the incidence of the event by the new treatment. In this article, we provide an exact method for calculating the sample size so that there will be adequate power to ensure that the exact upper confidence limit is below the threshold. Based on this we can design a study to achieve both objectives. PMID- 9741863 TI - Efficient clinical research. PMID- 9741862 TI - Three years of experience with prospective randomized effectiveness studies. AB - We developed methodology for prospective randomized effectiveness studies using a demonstration project at a multispecialty practice, health maintenance organization, and hospital in academic medical center. An operational unit called the effectiveness registry was developed to design and support comparisons of potential practice improvements with standard care. The studies differ from observational effectiveness studies in that they provide long-term follow-up of randomized comparison groups. Physician involvement in data collection is limited. No tests or observations are made other than those required for clinical care. Follow-up and data collection are modeled after tumor registry procedures. Patients who refuse randomization enter the study in whichever treatment arm they choose. The protocol for each study is approved by the institutional review board (IRB) before recruitment begins, and all patients, randomized and nonrandomized, sign an informed consent document. Between its beginning on October 7, 1993 and April 7, 1997, the IRB approved 14 trials. Four were terminated after entering at most a few patients. Recruitment is complete in four trials and continues in six. Randomization was accepted by 74% (596/804) of the patients. Over 800 patients in 10 studies are being followed at least annually. Major peer-reviewed journals have accepted reports of initial findings for two studies. Prospective randomized effectiveness studies are feasible in the multipractice setting and have potential to provide useful and reliable assessment of treatment outcomes. Collaborative arrangements between several institutions are needed to provide larger sample sizes. PMID- 9741864 TI - Recurrent miscarriage (REMIS) study: how should data from women who do not become pregnant be handled? AB - The Recurrent Miscarriage (REMIS) study is a double-blind, multicenter, randomized clinical trial designed to evaluate the efficacy of immunization with paternal leukocytes in the prevention of miscarriages in women who have had three or more unexplained pregnancy losses. Women entering the study are immunized with their husband's leukocytes or with a saline control before they become pregnant. After becoming pregnant, they receive weekly ultrasound examinations and psychological support during the first trimester and are followed until a successful delivery or a miscarriage occurs. The primary analysis for the study will be an intent-to-treat analysis in which we shall compare the proportion of successes in the two groups, defining a "success" as a pregnancy achieved within 12 months of randomization that results in a viable offspring. We shall count both miscarriages and nonpregnancies as failures, owing to the possibility of very early losses prior to the detection of pregnancy. In a secondary analysis, we shall exclude women who do not become pregnant within the alloted 12 month period. We compared the test size and power of these two approaches under various configurations for the true rates of nonpregnancy, miscarriage, and delivery in the two groups. Although the analysis excluding nonpregnant women achieves greater power for alternatives in which pregnancy rates are equal and live birth rates higher in the treated group, the rejection rate is not adequately controlled when pregnancy rates differ but live birth rates are unaffected by treatment. It can also lead to a reduction in power if the treatment prevents early as well as later losses. We conclude that the intent-to-treat analysis should remain the primary analysis for the trial. PMID- 9741865 TI - A design alternative for two-stage, phase II, multicenter cancer clinical trials. AB - Data are reviewed upon completion of patient accrual to the first stage of a two stage phase II cancer clinical trial and prior to initiating accrual on the second stage. Often accrual must be suspended while the data from the first stage are collected and evaluated. In an effort to maintain patient accrual momentum and avoid the logistical problems of suspending and reactivating a multicenter study, an alternative approach to conducting two-stage phase II studies, which allows patient accrual to continue while the results of the first stage are reviewed, is proposed. The properties of the proposed design are examined. PMID- 9741866 TI - Comparability of absolute/percent CD4+ T-lymphocytes completed locally and centrally. Hemophilia Growth and Development Study. AB - The Hemophilia Growth and Development Study (HGDS) is a multicenter longitudinal study of 333 male children and adolescents with moderate or severe hemophilia, ranging in age from 6 to 19 at entry. Sixty-two percent of the cohort was infected with human immunodeficiency virus (HIV) in the late 1970s and early 1980s through exposure to contaminated clotting factor concentrates. The HGDS has followed this cohort since 1989. HGDS subjects have blood drawn twice each year for t-lymphocyte subsets, with fresh blood shipped overnight to a central laboratory. T-lymphocyte subsets from the same blood draw are often determined locally as well. To evaluate interlaboratory variation, we examined the comparability of pairs of local and central results for CD4+ absolute counts and percents. Ninety-four pairs of absolute counts and 73 pairs of percent CD4 + results were available. We calculated concordance correlation coefficients, which evaluate the agreement between two readings from the sample by measuring the variation from the 45 degrees line through the origin. Absolute counts were square root transformed. Comparability of the pairs was high for both absolute counts and percents (0.93 and 0.92, respectively). Agreement was high whether we determined the CD4+ counts and percents centrally, using fresh samples received the day after the examination (0.95, 0.95), or from specimens that were frozen upon receipt and batched for later testing (0.90, 0.87). We conclude that when a centrally processed CD4+ result is unavailable because of shipping problems or loss of specimens, a study may reasonably accept a CD4+ result completed locally, if validity checks indicate good comparability. In the HGDS, the data provided by the local laboratories were of comparable quality to those provided by the central laboratories. PMID- 9741867 TI - Recruitment strategies in the women's health trial: feasibility study in minority populations. WHT:FSMP Investigators Group. Women's Health Trial:Feasibility Study in Minority Populations. AB - The Women's Health Trial:Feasibility Study in Minority Populations (WHT:FSMP) examined the feasibility of recruiting postmenopausal women from a broad range of racial and socioeconomic backgrounds into a primary prevention trial requiring marked reductions in dietary fat. Postmenopausal women aged 50-79 yr who had no history of cardiovascular disease or cancer and who consumed 36% or more total energy from fat qualified to participate. We randomized the women into dietary intervention (60%) or control (40%) groups; we aimed to randomize 750 women in 18 months in each of the three clinical centers. All centers achieved goals for randomization based on ethnicity, and two centers exceeded overall recruitment goals. The greatest source of randomized participants was mass mailing, followed by items in the media, referrals, and community outreach. Recruitment yields were generally similar for the ethnic groups but lower for less-educated participants. The experience of WHT:FSMP indicates that postmenopausal women from the African American, Hispanic, and non-Hispanic white communities can be recruited into dietary intervention studies for the prevention of disease. PMID- 9741868 TI - Guidelines for quality assurance in multicenter trials: a position paper. AB - In the wake of reports of falsified data in one of the trials of the National Surgical Adjuvant Project for Breast and Bowel Cancer supported by the National Cancer Institute, clinical trials came under close scrutiny by the public, the press, and Congress. Questions were asked about the quality and integrity of the collected data and the analyses and conclusions of trials. In 1995, the leaders of the Society for Clinical Trials (the Chair of the Policy Committee, Dr. David DeMets, and the President of the Society, Dr. Sylvan Green) asked two members of the Society (Dr. Genell Knatterud and Dr. Frank Rockhold) to act as co-chairs of a newly formed subcommittee to discuss the issues of data integrity and auditing. In consultation with Drs. DeMets and Green, the co-chairs selected other members (Ms. Franca Barton, Dr. C.E. Davis, Dr. Bill Fairweather, Dr. Stephen George, Mr. Tom Honohan, Dr. Richard Mowery, and Dr. Robert O'Neill) to serve on the subcommittee. The subcommittee considered "how clean clinical trial data should be, to what extent auditing procedures are required, and who should conduct audits and how often." During the initial discussions, the subcommittee concluded that data auditing was insufficient to achieve data integrity. Accordingly, the subcommittee prepared this set of guidelines for standards of quality assurance for multicenter clinical trials. We include recommendations for appropriate action if problems are detected. PMID- 9741870 TI - What is your diagnosis? Sagittal and a small dorsally displaced slab fracture of the radial carpal bone with secondary degenerative joint disease. PMID- 9741871 TI - Myasthenia gravis: lessons from the past 10 years. AB - Over the past 10 years, significant advances have been made in our understanding of acquired myasthenia gravis (MG) in companion animals. The broad spectrum of presenting clinical signs has been defined and an accurate and sensitive diagnostic test is available. Even with these advances, the mortality rate in dogs with acquired MG remains unacceptably high. While an understanding of the genetic basis for susceptibility to autoimmune disease has started to be developed, the trigger for the initiation of this disease is not known and a mechanism for specific suppression of the aberrant immune response against the acetylcholine receptor remains a mystery. PMID- 9741869 TI - Design of the PID Evaluation and Clinical Health (PEACH) Study. AB - This paper describes the PID Evaluation and Clinical Health Study (PEACH), a multicenter, randomized clinical trial designed to compare treatment with outpatient and inpatient antimicrobial regimens among women with pelvic inflammatory disease (PID). PEACH is the first trial to evaluate the effectiveness and cost-effectiveness of currently recommended antibiotic combinations in preventing infertility, ectopic pregnancy, chronic pelvic pain, recurrent PID, and other health outcomes. It is also the largest prospective study of PID ever conducted in North America. We describe the PEACH study's specific aims, study organization, patient selection criteria, conditions for exclusion, data collected upon entry, randomization and treatment, adherence measures, follow-up activities, quality-of-life measures, outcomes, and statistical analyses. In the first 11 months of enrollment (March 1996-January 1997), 312 women were randomized. Of eligible women, 59% consented to enroll. Participating women are primarily black (72%) and young (mean age 24 years). After a median of 5.5 months of follow-up, we were in contact with 95% of study participants. The PEACH study will provide a rationale for selecting between inpatient and outpatient antibiotic treatment, the two most common treatment strategies, for PID. PMID- 9741872 TI - Can owners assess outcome following treatment of canine cruciate ligament deficiency? AB - Forty-seven owners of 48 dogs surgically treated for cranial cruciate ligament deficiency were asked to assess the outcome of treatment at one year postsurgery. A standard visual analogue scale assessment tool was used in all cases and owners were asked to score a variety of generic and disease-specific features. The reliability and responsiveness of the assessment tool were estimated and found to be acceptable; generic features, such as 'level of disability', showed the greatest reliability and responsiveness. The results of assessments by owners were compared to assessments by a veterinary surgeon but few significant correlations were found. Assessments by owners appear to be a useful outcome measure in this disease but are unlikely to be directly comparable to assessments by a veterinary surgeon. PMID- 9741873 TI - Ultrasonographic evaluation of adhesions induced by incisional gastropexy in 16 dogs. AB - A study was undertaken to evaluate the use of ultrasonography to assess the gastropexy site for permanent adhesion in clinical cases. Two groups, each comprising eight dogs, were studied, all 16 cases undergoing decompression, anatomical repositioning of the stomach and an incisional gastropexy after gastric dilatation-volvulus (GDV). Group 1 was set up as a prospective group in which ultrasonographic examinations were performed three times (mean three, 12 and 67 days) after surgery to evaluate the gastropexy region. The gastropexy site was assessed ultrasonographically at only one stage (mean 449 days after surgery) in the group 2 dogs. Criteria used to assess the usefulness of the ultrasonographic evaluation included the ability to identify the gastropexy site, to obtain measurements of the length and thickness of the site and to assess the ultrasonographic appearance of the different gastric wall layers. The average number of peristaltic contractions and degree of gastric filling were also evaluated. The fixation between the stomach and the abdominal wall was easily detected in all 16 cases. Ultrasonography proved to be a simple and non-invasive technique to assess the permanency of the gastropexy. The incisional gastropexy was relatively easy to perform and induced permanent adhesions in all 16 dogs, without recurrence of GDV. PMID- 9741874 TI - Use of computed tomography to investigate cheek tooth abnormalities in chinchillas (Chinchilla laniger) AB - Computerised tomographic scanning was used to investigate tooth structure in chinchillas (Chinchilla laniger), both cheek tooth crown and root abnormalities being common in this species. This paper describes a common form of dental disease affecting species with continuously growing teeth, with particular reference to the chinchilla, and confirms the potential role of computed tomography (CT) in its early diagnosis. CT imaging is compared with previously available methods of investigation which frequently fail to detect early pathological changes. PMID- 9741875 TI - Using the Unger system to classify 386 long bone fractures in dogs. AB - A system already described by Unger and others was used to classify long bone fractures in dogs. The present paper reports experiences using the fracture classification system regarding its ease of use and the ability to analyse the data generated. Three hundred and eighty-six canine long bone fractures were classified from radiographs. Results were assessed by reviewing the medical records or by sending questionnaires to referring veterinarians. There were a few inconsistencies, particularly in classifying proximal ulnar fractures, but the system was easy to use and data retrieval was readily accomplished. Data from the system were used to compare the results of repairs of diaphyseal fractures of the radius/ulna, femur and tibia/fibula. A chi square analysis was used to determine significant differences between the outcome scores of the three fracture types. Fractures of the femoral diaphysis had a statistically poorer outcome than did diaphyseal fractures of the radius/ulna or tibia/fibula. PMID- 9741876 TI - A case of feline paraneoplastic alopecia with secondary Malassezia-associated dermatitis. AB - A 13-year-old neutered female domestic shorthaired cat had progressive ventral abdominal alopecia attributed initially to hyperthyroidism. Corrective treatment by unilateral thyroidectomy did not, however, resolve the dermatosis and the alopecia progressed to involve the whole ventral trunk, the lower limbs and the head. Pruritus of the lower limbs was a prominent feature and was associated with the finding of Malassezia on cytology; Malassezia-associated dermatitis was diagnosed. Resolution of pruritus was seen after treatment with oral ketoconazole and a cleansing shampoo to eliminate the yeast, but severe polyphagia, small intestinal diarrhoea and polydipsia developed subsequently and the cat was euthanased. Necropsy revealed an exocrine pancreatic adenocarcinoma with hepatic metastases. The pancreatic, hepatic and dermatological lesions were found to be typical of feline paraneoplastic alopecia (FPA). Malassezia-associated dermatitis can be associated with pruritus in cats with FPA. PMID- 9741877 TI - Arthrodesis of the shoulder for synovial osteochondromatosis. AB - A case of synovial osteochondromatosis (SOC) in a young deerhound's shoulder is reviewed. The age of onset and initially unaffected articular surfaces suggested primary SOC. Histological criteria for primary versus secondary osteochondromatosis are contradictory and unclear. Initial loose body removal and partial synovectomy resulted in several months of improvement but loose bodies returned. Severe degenerative changes were found one year after the initial exploratory surgery. Arthrodesis resulted in a functional, non-painful joint. PMID- 9741878 TI - MRI and electrophysiological abnormalities in a case of canine globoid cell leucodystrophy. AB - A six-month-old West Highland white terrier with progressive, multifocal neurological disease was diagnosed with canine globoid cell leucodystrophy (GCL). Magnetic resonance imaging (MRI) of the brain was performed, as well as electrophysiological testing (including brainstem auditory evoked response, peripheral nerve conduction velocity, repetitive stimulation, F wave analysis and electromyography). MRI findings were consistent with diffuse, symmetrical white matter disease. Electrodiagnostic testing revealed evidence of peripheral neuropathy and an abnormal brainstem auditory evoked response. These observations were consistent with the pathological changes in central and peripheral white matter described for canine GCL, and resembled what has been described in human patients. It is believed that the tests may raise the suspicion of GCL in dogs and may aid in monitoring disease progression. PMID- 9741879 TI - Heartworm disease. PMID- 9741880 TI - Protective role of Anacardium occidentale extract against streptozotocin-induced diabetes in rats. AB - The protective effect of Anacardium occidentale aqueous extract against streptozotocin-induced diabetes was evaluated in rats. The rats were treated with 175 mg/kg of the extract per os, twice daily; beginning 2 days before streptozotocin (STZ) injection. A total of 3 days after STZ administration, there was a 48% increase in blood glucose level in pretreated rats, compared with a 208% increase in diabetic control rats treated with STZ alone. Furthermore, these pretreated animals presented no glycosuria, a normal weight gain and a non significant increase in food and fluid intake at the end of the treatment compared with the normal control. Diabetic control animals showed a positive glycosuria, body weight loss, a real polyphagia and polydypsia. These results indicate the protective role of Anacardium occidentale extract against the diabetogenic action of STZ. PMID- 9741881 TI - Toxic effects of Erycibe obtusifolia, a Chinese medicinal herb, in mice. AB - Extract of stem of Erycibe obtusifolia (EO) at doses of 10, 20 and 30 mg/kg was experimentally tested through oral and intraperitoneal administration. Toxic effects of EO were assessed through functional changes of the liver and kidneys. Mice died immediately following the i.p. injection at the dose of 10 mg/kg. However, no death occurred after the oral administration at the dose of 10, 20 or 30 mg/kg under close observations for at least 2 weeks. Changes of several functional parameters in both the liver and kidney appeared simultaneously after the oral administration. Although the higher dose increased the levels of serum glutamate-oxalate-transaminase (sGOT), serum glutamate-pyruvate-transaminase (sGPT), and blood urea nitrogen (BUN), and decreased the levels of hematocrit at 6 h after the treatment, no distinct dose-dependent relationship existed between the administered doses and the changes in functional parameters observed. PMID- 9741882 TI - Plants used in Guatemala for the treatment of protozoal infections: II. Activity of extracts and fractions of five Guatemalan plants against Trypanosoma cruzi. AB - The activities of crude plant extracts of five plants popularly used in Guatemala against bacterial and protozoal infections and some of their fractions have been evaluated against the trypomastigote and epimastigote forms of Trypanosoma cruzi in vitro. The most active fraction of Neurolaena lobata has also been screened in vivo. Main in vitro activities against trypomastigotes have been observed for the hexane and ethanol extracts of N. lobata (Asteraceae). Both extracts were also active against epimastigotes, whereas all other extracts tested had no effect on epimastigotes. For the hexane extracts of Petiveria alliacea (Phytolaccaceae) and Tridax procumbens (Asteraceae) a marked inhibition of trypomastigotes has been found. Also the ethanol extracts of Byrsonima crassifolia (Malpighiaceae) leafs and Gliricidia sepium (Papilionaceae) bark showed some trypanocidal activity. Fraction 2 of the ethanol extract of N. lobata was highly active against T. cruzi as well in vitro as in vivo. The chloroforme fraction of P. alliacea showed a high inhibition of trypomastigotes in vitro. Also three fractions of the active extract of B. crassifolia inhibited T. cruzi trypomastigotes. No fraction of G. sepium bark extract showed a marked trypanocidal activity. PMID- 9741883 TI - Contraceptive and non-estrogenic effects of methanolic extract of Asparagus pubescens root in experimental animals. AB - The methanolic extract of Asparagus pubescens Bak root was investigated for its contraceptive activity in mice, rats and rabbits. The extract dose-dependently (0.5-1.5 g/kg) protected the animals from conception for 4-14 gestational periods in rabbits, rats and mice. It inhibited fetal implantation, as was confirmed by laparotomy on day 10 of pregnancy. The pups showed significant change in weight and length (P < 0.05-0.001) with 1.5 g/kg compared to the control fetal defects. In ovariectomized immature young rats and mice, there was a dose-dependent decrease in uterine wet weight (P < 0.001). The extract did not induce any uterotrophic effects or immature vaginal opening when compared to estrogen treated groups. Its contraceptive effect may in part be due to its anti implantation and/or a direct effect on the uterus. PMID- 9741884 TI - Evaluation of Nigerian traditional medicines: II. Effects of some Nigerian folk remedies on peptic ulcer. AB - Antiulcer activity of four medicinal plants, Diodia sarmentosa (whole plant), Cassia nigricans (leaves), Ficus exasperata (leaves) and Synclisia scabrida (leaves), which are commonly used by the Nigerian traditional healers for the treatment of peptic ulcer were investigated. Acute toxicity tests were also carried out. The results revealed that the four extracts possess significant anti ulcerogenic properties in a dose-dependent way. They protected rats from aspirin induced ulcerogenesis, delayed intestinal transit, increased the pH, and decreased both the volume and acidity of gastric secretion. These results correlate with local use of the plants. PMID- 9741885 TI - Inhibition of nitric oxide synthesis by butanol fraction of the methanol extract of Ulmus davidiana in murine macrophages. AB - Since there is increasing evidence that nitric oxide (NO) plays a crucial role in the pathogenesis of inflammatory diseases, this study was undertaken to address whether the methanol (MeOH) extract and its fractions of the bark of Ulmus davidiana Planch (Ulmaceae) could modulate the expression of inducible NO synthase (iNOS) in thioglycollate-elicited murine peritoneal macrophages and murine macrophage cell line, RAW264.7 cells. Stimulation of the peritoneal macrophages and RAW264.7 cells with interferon-gamma (IFN-gamma) and lipopolysaccharide (LPS) resulted in increased production of NO in the medium. However, the butanol (BuOH) fraction of the MeOH extract of U. davidiana barks showed marked inhibition of NO synthesis in a dose-dependent manner. The inhibition of NO synthesis was reflected in the decreased amount of iNOS protein, as determined by Western blotting. The BuOH fraction did not affect the viability of RAW264.7 cells, as assessed by methylthiazol-2-yl-2, 5-diphenyl tetrazolium bromide (MTT) assay; rather, it reduced endogenous NO-induced apoptotic cell death via inhibition of NO synthesis in RAW264.7 cells. On the other hand, the BuOH fraction showed no inhibitory effect on the synthesis of NO by RAW264.7 cells, when iNOS was already expressed by the stimulation with IFN-gamma and LPS. Collectively, these results demonstrate that the BuOH fraction inhibits NO synthesis by inhibition of the induction of iNOS in murine macrophages. PMID- 9741886 TI - Pharmacological, electrophysiological and toxicity studies of Limacia scanden Lour. (Menispermaceae). AB - Pharmacological studies showed that Limacia scanden Lour. extracts have sympathomimetic activities similar to noradrenaline (NA). A crude extract of Limacia scanden injected intravenously as a single bolus induced a dose-dependent increase in arterial blood pressure in anaesthetized rats and cats. Pretreatment with a non-specific alpha blocker phentolamine (10(-5) M) blocked this effect, whereas the beta blocker propanolol (10(-5) M) did not. The extract also reduced intestinal motility and this response could be blocked by pretreatment with phentolamine (10(-5) M) and specific alpha1-blocker, prazosin (10(-5) M). In superfused rabbit aorta preparations, it induced an increase in contractions. This effect was blocked by pretreatment with prazosin (10(-5) M), whereas the alpha2-blocker yohimbine (10(-5) M) had only a slight effect. The effects of NA on superfused aorta strip contraction were similar to extract. Toxic symptoms were manifested in less than 5 min when the mice were given 465 mg/kg of extract intraperitoneally. Physiological and behavioural changes observed in dying mice implicated serious malfunctioning of the autonomic nervous system and motor activity. Electrophysiological studies on the tonically autoactive neuron (TAN) of the snail Achantina fulica Ferussac revealed that crude extract of Limacia scanden induced excitatory responses which were similar to those of serotonin (5 HT) stimulation. Studies with different ionic compositions of the bathing saline revealed that this excitatory effect of Limacia scanden could be attributed either to release of endogenous serotonin or inhibition of 5-HT reuptake in the CNS. This observation could tentatively be used to provide the framework towards elucidating the mechanism and rationale for the use of this plant in traditional medicine in the treatment of depression and affective disorders. PMID- 9741887 TI - Medical ethnobotany of the Zapotecs of the Isthmus-Sierra (Oaxaca, Mexico): documentation and assessment of indigenous uses. AB - The Zapotec inhabitants of the Sierra de Oaxaca foothills (Mexico) live in an area of great botanic diversity. In daily subsistence and in response to illness, plants play a major role. An inventory of the Zapotec medicinal ethnobotany was carried out during 17 months of fieldwork. A total of 3611 individual responses concerning medicinal and non-medicinal uses for 445 different species of plants were documented. For the subsequent semi-quantitative analysis of data, the uses were grouped into ten categories and the responses for each species were summed up in each of these ten groups to yield rank-ordered lists. For the high rank ordered and, hence, culturally important species, an assessment of the therapeutic potential was conducted using ethnobotanical, phytochemical and pharmacological data in the literature. Studies confirming the attributed properties or a scientific explanation of therapeutic use, as well as toxicological data, are still lacking for many of these species. The quantitative approach described will be the basis for future studies on the pharmacology and phytochemistry of Zapotec medicinal species. Finally, these data should also serve as a basis for biodiversity conservation and community development. PMID- 9741888 TI - The inhibitory effect of common traditional anti-rheumatic herb formulas on prostaglandin E and interleukin 2 in vitro: a comparative study with Tripterygium wilfordii. AB - To understand the clinical efficacy of traditional anti-rheumatic herbal medicines on acute and severe arthritis or immune diseases, four herbal formulas and one herb were tested in vitro to determine their effects on prostaglandin E2 (PGE2) and interleukin 2 (IL2). Peripheral blood mononuclear cells from healthy subjects were incubated with different concentrations of four herbal formulas including Shaur Yau Gan Tsao Tang (SYGTT), Shang Jong Shiah Tong Yong Tong Feng Wan (SJSTY), Shu Jin Lih An Saan (SJLAS), Ma Shing Yih Gan Tang (MSYGT) and one herb, Tripterygium wilfordii (T2) with and without mitogen stimulation. PGE2 and IL2 from culture supernatant were measured by enzyme immunoassay. The results showed that SYGTT, SJSTY, SJLAS at concentration of 100 microg and MSYGT at 500 microg/ml can significantly inhibit PGE2 release (P < 0.05) from mononuclear cells. However, T2 at 2 microg/ml expressed the same response. For the inhibition of IL2, the concentration of SYGTT, SJSTY and SJLAS must exceed 100 symbol microg/ml. MSYGT failed to inhibit IL2 at even concentrations of 500 microg/ml but T2 at a very low concentration (0.6 microg/ml) could strongly inhibit it. The findings suggest that the majority of traditional anti-rheumatic herbal formulas or herbs, except for T2, should not be used to treat acute and critical arthritis or immune diseases. PMID- 9741889 TI - Screening of 34 Indian medicinal plants for antibacterial properties. AB - A total of 34 plant species belonging to 18 different families, selected on the basis of folklore medicinal reports practised by the tribal people of Western Ghats, India, were assayed for antibacterial activity against Escherichia coli, Klebsiella aerogenes, Proteus vulgaris, and Pseudomonas aerogenes (gram-negative bacteria) at 1000-5000 ppm using the disc diffusion method. Of these 16 plants showed activity; among them Cassia fistula, Terminalia arjuna and Vitex negundo showed significant antibacterial activity against the tested bacteria. Our findings confirm the traditional therapeutic claims for these herbs. PMID- 9741890 TI - Screening of some Indian medicinal plants for their antimicrobial properties. AB - A total of 82 Indian medicinal plants traditionally used in medicines were subjected to preliminary antibacterial screening against several pathogenic and opportunistic microorganisms. Aqueous, hexane and alcoholic extracts of each plant were tested for their antibacterial activity using agar well diffusion method at sample concentration of 200 mg/ml. The results indicated that out of 82 plants, 56 exhibited antibacterial activity against one or more test pathogens. Interestingly, extracts of five plants showed strong and broad spectrum activity as compared to rest of 51 plant extracts which demonstrated moderate activity. On the whole the alcoholic extracts showed greater activity than their corresponding aqueous and hexane extracts. Among various extracts, only alcoholic extracts of Emblica officinalis, Terminalia chebula, Terminalia belerica, Plumbago zeylanica and Holarrhena antidysenterica were found to show potentially interesting activity against test bacteria. These active crude alcoholic extracts were also assayed for cellular toxicity to fresh sheep erythrocytes and found to have no cellular toxicity. PMID- 9741891 TI - Computerised controllers for safety critical medical applications. AB - A novel design for a computer control system to be employed in safety critical applications, as found in medical environments, is presented. It features a low complexity, fault detecting hardware architecturally supporting a strictly cyclic operating mode, as known from programmable logic controllers, and a specification level, graphical programming technique based on the interconnection of application oriented standard software function modules. By design, there is no semantic gap between the programming and machine execution levels. Thus enabling the safety licensing of application software by an extremely simple but rigorous method, the software verification problem is satisfactorily solved for a large application area, where failures may cause hazards to, or even loss of, human safety and lives. PMID- 9741892 TI - Enhancement and associative restoration of electronic portal images in radiotherapy. AB - Electronic portal imaging devices use the high energy treatment beam to project the body interior of the patient during radiation onto a fluorescent screen that is scanned by a camera. Because of the imaging physics, the unprocessed images of very poor quality, but they are the only available information during treatment for observation of the patient's organs. This paper presents an approach that combines an associative restoration algorithm with a fuzzy image enhancement technique. By fusion of the electronic portal image (EPI) with a pre-treatment captured simulator image (SI) a higher image quality than by conventional techniques is achieved. PMID- 9741893 TI - The Creation of a global telemedical information society. AB - Healthcare is a major candidate for improvement in any vision of the kinds of 'information highways' and 'information societies' that are now being visualized. The medical information management market is one of the largest and fastest growing segments of the healthcare device industry. The expected revenue by the year 2000 is US$21 billion. Telemedicine currently accounts for only a small segment but is expanding rapidly. In the USA more than 60% of federal telemedicine projects were initiated in the last 2 years. The concept of telemedicine captures much of what is developing in terms of technology implementations, especially if it is combined with the growth of the Internet and World Wide Web (WWW). It is foreseen that the World Wide Web (WWW) will become the most important communication medium of any future information society. If the development of such a society is to be on a global scale it should not be allowed to develop in an ad hoc manner. For this reason, the Euromed Project has identified 20 building blocks resulting in 39 steps requiring multi-disciplinary collaborations. Since, the organization of information is therefore critical especially when concerning healthcare the Euromed Project has also introduced a new (global) standard called 'Virtual Medical Worlds' which provides the potential to organize existing medical information and provide the foundations for its integration into future forms of medical information systems. Virtual Medical Worlds, based on 3D reconstructed medical models, utilizes the WWW as a navigational medium to remotely access multi-media medical information systems. The visualization and manipulation of hyper-graphical 3D 'body/organ' templates and patient-specific 3D/4D/and VR models is an attempt to define an information infrastructure in an emerging WWW-based telemedical information society. PMID- 9741894 TI - Mechanical imaging: a new technology for medical diagnostics. AB - Mechanical imaging (MI) is a newly developed modality of medical diagnostics based on reconstruction of tissue structure and viscoelastic properties using mechanical sensors. The essence of MI is the solution to an inverse problem using the data of stress patterns on the surface of tissue compressed by a pressure sensor array. Imaged tissue structures are presented in terms of their viscoelastic properties. Evaluation of tissue 'hardness' (shear elasticity modulus) provides a means for characterizing the tissue, differentiating normal and diseased conditions and detecting tumors and other lesions. In contrast to the other existing methods of medical imaging which use sophisticated hardware such as superconductive magnets, expensive X-ray equipment and complex ultrasonic phased arrays, MI hardware consists of inexpensive mechanical sensors and a positioning system connected to a PC. A key feature of MI is 'knowledge-based imaging'. To produce a three-dimensional image, the computer uses both the measured parameters of an individual examined object and a general database on anatomy and pathology of the object. Two applications of MI are currently being developed: MI for mass screening and detection of breast cancer and MI for imaging the prostate and diagnosing prostate diseases. A prototype of the device for mechanical imaging of the prostate has been developed and is being tested clinically at the Robert Wood Johnson Medical School, New Jersey. The device is comprised of a transrectal probe with a position sensor and a pressure sensor array mounted on the articulated tip, an electronic unit and a PC. Results of extensive laboratory studies with rubber prostate models and initial data obtained in clinical trials strongly suggests that for certain applications the MI technology, as a new modality of imaging, has a diagnostic potential comparable to that of conventional diagnostic technologies. Mechanical imaging of the prostate appeared to be an efficient means of objectively evaluating and imaging the prostate and detecting prostate cancer. PMID- 9741895 TI - The decision support system for telemedicine based on multiple expertise. AB - This paper discusses the application of artificial intelligence in telemedicine and some of our research results in this area. The main goal of our research is to develop methods and systems to collect, analyse, distribute and use medical diagnostics knowledge from multiple knowledge sources and areas of expertise. Use of modern communication tools enable a physician to collect and analyse information obtained from experts worldwide with the help of a decision support medical system. In this paper we discuss a multilevel representation and processing of medical data using a system which evaluates and exploits knowledge about the behaviour of statistical diagnostics methods. The presented technique is able to acquire semantically-essential information from the complex dynamics of quasi-periodical medical signals by applying recursively-ordinary statistical tools. A method and an algorithm are elaborated to select automatically the most appropriate diagnostics method for each case under consideration. We suggest the use of a voting-type technique to search for consensus among the different opinions of medical experts. Research results can be applied in the development of a telediagnostics expert medical system and medical teleconsulting support system. PMID- 9741896 TI - Cellular automata and follicle recognition problem and possibilities of using cellular automata for image recognition purposes. AB - Cellular automata are discrete dynamical systems whose behaviour is completely specified in terms of a local relation. Guided by a suitable recipe, they can simulate a whole hierarchy of structures and phenomena. While investigating the problem of follicle recognition in ultrasonic images of women's ovaries, we became increasingly interested in using cellular automata for this purpose. We were very successful, which encouraged us to further investigate the use of cellular automata for image recognition purposes in general. This paper presents the results of our research in this area, along with the details of how we solved the follicle recognition problem. PMID- 9741897 TI - Relating clinical and neurophysiological assessment of spasticity by machine learning. AB - Spasticity following spinal cord injury (SCI) is most often assessed clinically using a five-point Ashworth score (AS). A more objective assessment of altered motor control may be achieved by using a comprehensive protocol based on a surface electromyographic (sEMG) activity recorded from thigh and leg muscles. However, the relationship between the clinical and neurophysiological assessments is still unknown. In this paper we employ three different classification methods to investigate this relationship. The experimental results indicate that, if the appropriate set of sEMG features is used, the neurophysiological assessment is related to clinical findings and can be used to predict the AS. A comprehensive sEMG assessment may be proven useful as an objective method of evaluating the effectiveness of various interventions and for follow-up of SCI patients. PMID- 9741898 TI - Mass transport enhancement by ultrasound in non-degradable polymeric controlled release systems. AB - In this work, an attempt was made to characterize mass transport enhancement in non-erodible polymeric matrices, caused by ultrasound. It was found that drug release rates from polymeric matrices exposed to ultrasound, can be controlled by modifying parameters like: ultrasound frequency, molecular weight of the incorporated drug and structure of the polymeric matrix (size of pores in the network). It is suggested that the enhancing effect of ultrasound on drug release from non-erodible polymers is due to the contribution of a convective term, generated by cavitation, without any destructive effect on morphology of the polymer. This phenomenon was found to be more pronounced in systems which are mass-transport limited. PMID- 9741899 TI - Controlled iontophoretic release of glucocorticoids through epithelial cell monolayers. AB - In the present study the iontophoretic transdermal delivery of three different glucocorticoids through a confluent monolayer of MDCK cells, mimicking biological barriers, was studied. For this experiment an in vitro model with platinum electrodes for iontophoresis and MDCK cells was developed. With this model investigations concerning the biocompatibility of the cells depending on different current densities and the iontophoretic permeation of the three glucocorticoids through the cell monolayer were carried out. The permeation behavior of this living biological barrier should be very similar to the non living barrier, human stratum corneum. Different current densities (12.74-38.22 microA/cm2) and a pulsatile application of continuous current to the cell monolayer were investigated. The pulsatile application of the current resulted in a step-like permeation profile. In all cases the electric current induced an reversible increase of the porosity of the cell monolayer demonstrated by transepithelial electrical resistance measurements and by sodium fluorescein assay. PMID- 9741900 TI - Polymeric nanoparticles as delivery system for influenza virus glycoproteins. AB - The objective of this work was to develop a new delivery system which could enhance the mucosal immune response to influenza virus antigens. Poly(D,L-lactide co-glycolide) nanoparticles of about 200 nm containing hemagglutinin were chosen as the delivery system. Due to the amphiphilic nature of hemagglutinin (hydrophilic HA1 and hydrophobic HA2), nanoparticles were prepared by both classical oil in water solvent evaporation technique as well as by a [(water-in oil) in water] solvent evaporation technique. Hemagglutinin was well encapsulated in nanoparticles prepared by both techniques. Molecular weight and antigenicity of entrapped hemagglutinin were not affected by the entrapment procedure. PMID- 9741901 TI - Pharmacodynamic and pharmacokinetic rationales for the development of an oral controlled-release amoxicillin dosage form. AB - The goal of this investigation was to develop an oral sustained-release formulation for amoxicillin that would maximize the duration of active drug concentration in the extracellular fluid, thus increasing the dosing interval while assuring antimicrobial activity. This rationale is based on the pharmacodynamic properties of the drug which is non- concentration dependent on the one hand, while requiring long exposure of the pathogen to the drug with minimal post-antibiotic effect on the other. Due to pharmacokinetic constraints, including short biological half-life and limited 'absorption window' (confined to the small intestine) with poor colonic absorption, the new matrix tablet formulation, composed of hydrophilic (hydroxypropyl methyl-cellulose) polymer, was designed to release 50% of its contents within the first 3 h and to complete the drug release process over 8 h (under in vitro conditions). The pharmacokinetics of the new formulation was evaluated in 12 healthy volunteers and compared to a conventional gelatin capsule with both formulations containing 500 mg amoxicillin. The plasma concentrations of active amoxicillin and penicilloic acid were determined by an HPLC method with a fluorometric detector. It was found that the area under the concentration-time curve and maximal serum amoxicillin concentrations following the sustained release preparation were lower than the immediate release formulation. However, the time over the required threshold concentrations, i.e. the minimal inhibitory concentration (MIC) as well as the more clinically relevant parameter--four times MIC of the drug against susceptible pathogens, was found to be maintained for significantly longer periods. The results suggest that in order to achieve a twice daily dosing regimen that will provide therapeutic concentrations for the whole 12 h dosing intervals, a larger dose of the new formulation should be given (e.g. 750 mg or even 1 g twice daily). This recommendation is based on the large interindividual differences of the extent of amoxicillin absorption found in this investigation, and is intended to assure that the 'poor' absorbers will also benefit from full antibiotic efficacy. This dosing regimen will lead to increased patient compliance and improved therapeutic outcome. PMID- 9741902 TI - Polyethylene glycol-grafted poly-L-lysine as polymeric gene carrier. AB - A new series of gene carriers, polyethylene glycol (PEG)-grafted poly-L-lysine (PLL, mol. wt. = 25000) with three different PEG-grafted ratios (5, 10 and 25 mole%, which means 5, 10 and 25% of epsilon-amino group of PLL was modified by PEG), was synthesized. These new gene carriers, named comb-shaped PEG-g-PLL copolymer, showed a 5- to 30-fold increase in transfection efficiency compared to PLL alone on a human carcinoma cell line. It is likely that Hep G2 cells were transfected by plasmid DNA/PEG-g-PLL complexes through an endocytosis mechanism due to the fact that chloroquine increased transfection efficiency. Although Lipofectin, a cationic lipid formulation, showed slightly higher transfection efficiency than PEG-g-PLL in Hep G2 cells, our designed PEG-g-PLL demonstrated lower cytotoxicity, early gene expression and maintenance of gene expression for up to 96 h. PMID- 9741903 TI - Ovalbumin peptide encapsulated in poly(d,l lactic-co-glycolic acid) microspheres is capable of inducing a T helper type 1 immune response. AB - An ovalbumin (OVA) peptide, consisting of residues 323-339, was incorporated into poly(d,l lactic-co-glycolic acid) (PLGA) microspheres and administered to mice. It was hypothesized that microencapsulation of the peptide in PLGA microspheres would avoid the need for traditional adjuvants and bias the immune response towards a type 1 T helper (Th1) response. An immunomodulator, monophosphoryl lipid A (MPLA), was incorporated into the microspheres to determine its efficacy in enhancing a Th1 response. The specificity of the immune response was determined using a T cell proliferation assay. The type of T helper response was determined by analysis of the cytokine secretion profiles of the proliferating T cells. Following s.c. immunization, the results revealed a T cell-specific immune response for the encapsulated OVA peptide both with and without MPLA. The cytokine profiles revealed high levels of IFN-gamma with very low levels of IL-4 and IL-10, suggesting a Th1 response. Furthermore, incorporation of MPLA in the peptide loaded PLGA microspheres resulted in an increase in the production of IFN gamma. Hence, peptide-loaded PLGA microspheres are capable of eliciting a specific Th1 immune response, which may be further enhanced in the presence MPLA. PMID- 9741904 TI - Gene transfer vectors based on Sendai virus. AB - A gene delivery system is a fundamental technology used in human gene therapy. In order to treat patients suffering from incurable metabolic diseases, we must be able to deliver genes efficiently in situ and induce stable gene expression in non-dividing tissue cells. However, none of the current gene transfer systems (both viral and non-viral) satisfies this goal. In order to develop a novel gene delivery system that is free from the defects of existing gene transfer vectors, we analyzed natural biological phenomena that involve gene transfer and expression, and made artificial components that mimic the functioning of these systems. Our recent results shed light on three major aspects of gene transfer and expression: (1) the direct delivery of DNA into cytoplasm using fusogenic liposomes, (2) the transfer of DNA from cytoplasm to nucleus with a nuclear localization signal, and (3) the stabilization of DNA in the nucleus as an independent replicon. The possible development of a hybrid vector by combining these components is discussed. PMID- 9741905 TI - Crystalline properties of carbamazepine in sustained release hydrophilic matrix tablets based on hydroxypropyl methylcellulose. AB - The influence of hydroxypropyl methylcellulose (HPMC) on the crystal habit properties of carbamazepine in sustained release matrix tablets and in aqueous solutions was investigated using differential scanning calorimetry (DSC), X-ray powder diffraction and scanning electron microscopy (SEM). The results suggest that HPMC inhibits the transformation of carbamazepine to carbamazepine dihydrate in the gel layer of hydrated tablets and in aqueous solutions (depending on HPMC concentration), participates in its crystallization process and induces amorphism of carbamazepine crystals. The mechanism which explains these effects envisages the polymer serving as a template or microsubstrate for nucleation in the crystallization process. We assume that the interaction between the drug and polymer occurs by hydrogen bonding. The hydroxyl groups of the polymer may attach to the drug at the site of water binding, and thus its transformation to the dihydrate form, is inhibited. A more specific interaction involves structural matching (similar bond spacing distances) between inter-atomic distances in the crystal lattice of carbamazepine dimer and intra-atomic distances along the polymer chain. PMID- 9741906 TI - Solid-state stability assessment of controlled release tablets containing Carbopol 971P. AB - A tablet formulation containing dyphylline, Carbomer, magnesium stearate, talc and lactose was prepared by the direct compression method. The objective of the study was to assess the stability of these tablets after subjecting them to exaggerated conditions of temperature (4, 25, 37, 45 and 55 degrees C) and humidity (37 degrees/11%RH, 37 degrees/51%RH and 37 degrees C/91%RH). The samples were analyzed for crystalline, thermal and dissolution changes with time for a period of 12 months using X-ray diffractometry (XRD), differential scanning calorimetry (DSC) and U.S.P. Dissolution Apparatus 2. Moisture sorption studies of all tablets indicated sorption of large amounts of moisture at 37 degrees C/91%RH. The fit factors, f1 andf2, were calculated and used to compare the dissolution profiles. The results showed increased f1 and decreased f2 values at higher humidity, while the samples were fairly stable at lower humidity and at all temperatures studied. Powder XRD patterns of tablets showed a change in crystallinity at higher humidity. The thermal and crystalline data indicated that the change in dissolution behavior at higher humidity may be due to the conversion of dyphylline to its hydrate form. PMID- 9741907 TI - Azocrosslinked poly(acrylic acid) for colonic delivery and adhesion specificity: in vitro degradation and preliminary ex vivo bioadhesion studies. AB - This study reports on the performance of a novel polymeric material that is capable of providing site specificity in active agent delivery and the development of mucoadhesive interactions. Azo-networks, based on an acrylic backbone crosslinked with 4,4'-divinylazobenzene, were subjected to in vitro degradation and mucoadhesion (before and after degradation) testing in order to model their performance in the gastrointestinal tract. Advanced surface characterisation techniques (SEM, AFM, FTIR microscopy) were used to examine the network morphology prior to, and after degradation. The data obtained from these studies indicate that there is an optimum crosslinking density to allow non adhesive particles to reach the colon. Within the colonic environment, the azo network degrades to produce a structure capable of developing mucoadhesive interactions with the colonic mucosa. PMID- 9741908 TI - Nerve growth factor delivery systems. AB - Growth factors encourage tissue regeneration and differentiation, accelerate wound healing, and modulate neural repair. Thus, growth factor administration may become a useful treatment for neurodegenerative diseases, such as Alzheimer's disease or Parkinson's disease, which are characterized by the degeneration of neuronal cell populations. Controlled-release polymer delivery systems may be an important technology in enabling the prevention of neuronal degeneration, or even the stimulation of neuronal regeneration, by providing a sustained release of growth factors to promote the long-term survival of endogenous or transplanted cells. PMID- 9741909 TI - Crosslinked dextran--a new capsule material for colon targeting of drugs. AB - In the work presented here we have studied the application of glutaraldehyde crosslinked dextran as a capsule material for colon-specific drug delivery. A reaction mixture containing dextran, MgCl2, glutaraldehyde and polyethyleneglycol 400 in water was applied onto molding pins of nylon producing capsule caps and bodies. The capsule materials were characterized by measuring of the mechanical strength in compression and equilibrium degree of swelling. Based on these results an optimal composition for the capsule material was selected. The dextran capsules were loaded with hydrocortisone and subsequently, drug release was studied. The release was found to be about 10% during the initial 3 h in a buffer solution. Over a period of 24 h the release was about 35%. However, when the dextran capsules were challenged with a dextranase solution, simulating the arrival of the drug delivery system to the colon, the capsules quickly broke and the drug was released as a dose dump. The study shows that the dextran capsules are promising candidates for providing a colon-specific drug delivery. PMID- 9741910 TI - Antibody transport in cultured tumor cell layers. AB - This review summarizes our recent in vitro studies of the factors affecting the tumor penetration of immunoconjugates. The studies were designed to probe the mechanisms of diffusion and convection, using a cultured layer of mouse melanoma cells as a model tumor cell layer and an antibody to the murine transferrin receptor as a model ligand. Transport of the binding antibody was observed to be slower than that of a non-binding control, a result that is consistent with the "binding site barrier" hypothesis (Fujimori et al., J. Nucl. Med., 31: 1191-1198, 1990). Internalization of the antibody/receptor complex was necessary for this effect to be observed, implying that intracellular trafficking is a determinant of net tumor transport rates. Convective fluid flow exhibited a dependence on cell density that is consistent with a Poiseuille flow model, suggesting that convective transport occurs as laminar flow in tortuous channels. Implications for immunoconjugate therapy, limitations of the approach, and future directions of the research program are discussed. PMID- 9741911 TI - Targetable HPMA copolymer-adriamycin conjugates. Recognition, internalization, and subcellular fate. AB - Recognition, internalization, and subcellular trafficking of N-(2 hydroxypropyl)methacrylamide (HPMA) copolymer conjugates containing N-acylated galactosamine (GalN) or monoclonal OV-TL16 antibodies (Ab) have been investigated in human hepatocarcinoma HepG2 and ovarian carcinoma OVCAR-3 cells, respectively. The intrinsic fluorescence of fluorescein or adriamycin (ADR) attached to HPMA copolymers permitted us to follow the subcellular fate of HPMA copolymer conjugates by confocal fluorescence microscopy and fluorescence spectroscopy. The pattern of fluorescence during incubation of HPMA copolymer-ADR-GalN conjugate containing lysosomally degradable tetrapeptide (GFLG) side-chains with HepG2 cells was consistent with conjugate recognition, internalization, localization in lysosomes, followed by the release of ADR from the polymer chains and ultimately diffusion via the cytoplasm into the cell nuclei. A similar pattern was observed in OVCAR-3 cells for Ab targeted HPMA copolymer conjugates. To test our hypothesis that HPMA-copolymer-bound anticancer drugs will be inaccessible to the energy-driven P-glycoprotein efflux pump in multidrug resistant (MDR) cells, we have compared the internalization of the HPMA copolymer-ADR conjugates by sensitive (A2780) and ADR-resistant (A2780/AD) ovarian carcinoma cell lines. Preliminary data on relative retention of ADR in MDR (A2780/AD) cells indicate a higher intracellular ADR concentration after incubation with HPMA copolymer-ADR conjugate when compared to incubation with free (unbound) ADR. PMID- 9741912 TI - Folate-mediated targeting of antineoplastic drugs, imaging agents, and nucleic acids to cancer cells. AB - The receptor for the vitamin, folic acid, is overexpressed on a number of human tumors, including cancers of the ovary, kidney, uterus, testis, brain, colon, lung, and myelocytic blood cells. Conjugates of folic acid linked via its gamma carboxyl to either a single drug molecule or assembly of molecules can bind to and enter receptor-expressing cancer cells via folate receptor-mediated endocytosis. Because the affinity of folate conjugates for cell surface folate receptors is high (KD approximately 10(-10) M), folic acid derivatization allows the selective delivery of diagnostic and therapeutic agents to cancer cells in the presence of normal cells. This review will summarize studies aimed at folate mediated targeting of protein toxins, imaging agents, antisense oligodeoxynucleotides, genes, and liposomes specifically to cancer cells in vitro and in vivo. PMID- 9741913 TI - Delivery of molecular and cellular medicine to solid tumors. AB - To reach cancer cells in a tumor, a blood-borne therapeutic molecule or cell must make its way into the blood vessels of the tumor and across the vessel wall into the interstitium, and finally migrate through the interstitium. Unfortunately, tumors often develop in ways that hinder each of these steps. Our research goals are to analyze each of these steps experimentally and theoretically, and then integrate the resulting information in a unified theoretical framework. This paradigm of analysis and synthesis has allowed us to obtain a better understanding of physiological barriers in solid tumors, and to develop novel strategies to exploit and/or to overcome these barriers for improved cancer detection and treatment. PMID- 9741914 TI - Tissue-level transport mechanisms of intraperitoneally-administered monoclonal antibodies. AB - Monoclonal antibodies (MAbs), produced for specific tumor antigens, can be linked with radioisotopes or metabolic toxins and administered intraperitoneally (i.p.) to treat metastatic cancer located on the peritoneum. Despite their specific binding properties, these proteins distribute to the serosal surface of all tissues surrounding the cavity in the same manner as other serum proteins. Recent data have raised a problem of access of the solution containing the MAb to significant portions of the peritoneal surface. If the MAb does arrive at the surface of the tumor, it penetrates via diffusion and convection. The rapidity and depth of penetration of the MAb are very dependent on the binding characteristics of the MAb to the tumor cells. Current data indicate that tumors often have a large interstitial space relative to normal muscle, and this can accelerate both diffusion and convection. However, a highly permeable tumor vasculature in the absence of lymphatic drainage has also been shown to produce interstitial pressure gradients from the center toward the periphery of the tumor, setting up a potential outward flow which may be a significant barrier to the movement of MAbs into the nodule. While theoretical mechanisms of diffusion, convection, and binding are well established, there is still a great need for in vivo data. PMID- 9741915 TI - Targeting tumor cells with bispecific antibodies and T cells. AB - It has been known for some time that mammalian immune systems are capable of eliminating large tumor burdens. Redirecting the immune response of a patient to an established tumor has now become the focus of various therapeutic strategies. In this report, two projects toward this goal are described. The first project involves the development of a transgenic mouse model for T cell directed therapeutics. These mice express specific T cell receptor alpha and beta transgenes on a background in which the recombinational-activating-gene-1 (RAG) has been knocked out. The mice express cytotoxic T cells but not either T helper cells or B cells. Despite these deficiencies, the animals are capable of eliminating tumors that express the appropriate peptide/major histocompatibility complex ligand that is recognized by the alphabeta transgenic T cell receptor. Human tumors grow as transplants in these mice, thereby allowing various agents that redirect the endogenous T cells against human tumors to be tested. The second project involves a description of such agents: bispecific antibodies that simultaneously bind to an immune effector cell and a tumor cell. The bispecific antibody described here consists of folate attached to anti-T cell receptor antibodies, or their fragments. A single-chain Fv coupled with folate can redirect the lysis of human tumor cells that bear the high affinity folate receptor. Preliminary in vivo data showed that the folate/antibody conjugates were also capable of mediating rejection of the human tumor. This transgenic mouse model should now allow the evaluation and optimization of bispecific agents that can redirect a patient's own T cell response. PMID- 9741916 TI - Something new in the field of PLA/GA bioresorbable polymers? AB - Polymers issued from glycolic acid and lactic acids (PLAGA) are now used worldwide as bioresorbable devices in surgery and in pharmacology. Their abiotic hydrolytic degradation has been shown to depend on diffusion-reaction phenomena and to proceed homogeneously or heterogeneously, depending on many factors. Two initiators are presently used industrially to make PLAGA polymers by ring opening polymerisation of lactide and/or glycolide in the bulk, namely Sn octanoate and zinc metal. In this contribution, attention is paid to the differences generated by the use of these two initiator systems in the case of the polymerisation of DL lactide. Various poly(DL-lactide)s were prepared and characterised by size exclusion chromatography (SEC), differential scanning calorimetry (DSC) and nuclear magnetic resonance spectroscopy (NMR). These polymers were allowed to age in pH=7.4 isoosmolar phosphate buffer at 37 degrees C. Under these conditions, polymers prepared by the two initiator systems showed dramatic differences when the fates of parallel sided specimens of rather large dimensions were considered. These differences were related to the esterification of some of the OH chain ends by octanoic acid and to the presence of rather hydrophobic low molecular weight by-products which were insoluble in the solvent generally used to purify the crude PLAGA polymers. These new findings should be of great interest in the case of PLAGA based matrices aimed at drug delivery. PMID- 9741917 TI - Controlled chemical modification of hyaluronic acid: synthesis, applications, and biodegradation of hydrazide derivatives. AB - Controlled modification of the carboxylic acid moieties of hyaluronic acid with mono- and polyfunctional hydrazides leads to biochemical probes, biopolymers with altered physical and chemical properties, tethered drugs for controlled release, and crosslinked hydrogels as biocompatible scaffoldings for tissue engineering. Methods for polyhydrazide synthesis, for prodrug preparation, for hydrogel crosslinking, and for monitoring biodegradation are described. PMID- 9741918 TI - In-situ self-assembling protein polymer gel systems for administration, delivery, and release of drugs. AB - Sequential block copolymers consisting of tandem repetition of amino acids have been constructed and genetically produced based on the natural repeating structures of silk and elastin protein. Combinations of silklike and elastinlike amino acid sequence blocks in a high molecular weight protein polymer are used to confer properties similar to those observed with hard block and soft block segmented polyurethanes. A certain subset of these silk-elastinlike protein compositions, termed ProLastins, will undergo an irreversible solution to gel transition in physiological, aqueous solution. The transition occurs over time and can be controlled by temperature, solution conditions, and additives which either prevent or promote hydrogen bond-mediated chain crystallization. The process involves no covalent crosslinking. Characterization of the gelling properties of various ProLastin compositions and their ability to release compounds which are incorporated directly into the gels are presented. PMID- 9741919 TI - Effect of molecular architecture of hydrophobically modified poly(N isopropylacrylamide) on the formation of thermoresponsive core-shell micellar drug carriers. AB - Terminal-incorporation of hydrophilic or hydrophobic groups dramatically influences the phase transition of poly(N-isopropylacrylamide) (PIPAAm) because of a critical role of the polymer chain ends in initiation of the phase transition. Incorporation of an amino or hydroxyl group to one end of PIPAAm remarkably raised the LCST (lower critical solution temperature) and slowed down the rate of the phase transition, and these effects were more pronounced as the mole fraction of hydrophilic groups increased compared to the random copolymers of PIPAAm and hydrophilic co-monomers, such as acrylic acid (AAc) or dimethylacrylamide (DMAAm). Hydrophilic effects were more remarkable for hydroxyl groups, due to stronger hydrogen bonding with water. Terminal-modification (hydrophobization) was also more effective in producing hydrophobic effects on the PIPAAm phase transition in comparison with PIPAAm copolymers that were randomly modified along the main chain with hydrophobic co-monomers. Moreover, terminal-located hydrophobic groups were able to form hydrophobic microdomains that were clearly isolated from PIPAAm chains in aqueous media by the aggregation of hydrophobic segments. As a result, the obtained micellar aqueous solution showed the same LCST as pure PIPAAm, while the PIPAAm random copolymer with hydrophobic co-monomers formed incompletely separated microdomains. The LCST for this random copolymer was reduced with increasing hydrophobic co-monomer mole fraction. Hydrophobically terminal-modified PIPAAm produced thermo-responsive core-shell structures that exhibited the same LCST and the same thermal response rate as those of free linear PIPAAm chains. Such polymeric micellar structures show reversible thermoresponsive aggregation/dispersion and deformation/reformation in heating/cooling cycles through the LCST for pure PIPAAm. These properties indicate the possibility of using such a system as a thermoresponsive drug carrier with double targeting mechanisms, in both passive and active manners. PMID- 9741920 TI - Polymeric micelles for drug delivery: solubilization and haemolytic activity of amphotericin B. AB - Polymeric micelles may serve as nanoscopic, long-circulating carriers of hydrophobic drugs. In this study, we have researched the solubilization of amphotericin B (AmB), an antifungal drug, by micelles of poly(ethylene oxide) block-poly(beta benzyl-L-aspartate) (PEO-PBLA), the properties of the AmB-loaded PEO-PBLA micelles and the resultant haemolytic activity of AmB. AmB loading takes place during self assembly of PEO-PBLA micelles, and this occurs through a dialysis procedure as an alkaline aqueous solution replaces the selective solvent for the polymer and the drug. In this way, AmB reaches levels of 57 to 141 microg/ml, corresponding to a loading efficiency of 27-30% (loaded AmB/initial amount of AmB). The molar ratio of AmB to PEO-PBLA is 0.40 to 1.0. Pictures by transmission electron microscopy reveal spherical AmB-loaded PEO-PBLA micelles with a mean diameter of 25.8+/-4.2 nm. AmB-loaded PEO-PBLA micelles are nonhaemolytic at an AmB level of 10 microg/ml as assessed by release of haemoglobin, measured by UV-Vis spectroscopy. AmB as Fungizone, its standard formulation, completely lyses red blood cells at a level of 3.0 microg/ml in 30 min. In contrast, there is no haemolysis at 5.5 h for AmB-loaded PEO-PBLA micelles at 3.0 microg/ml of AmB, indicating the gradual release of AmB from PEO PBLA micelles. PEO-PBLA itself is nonhaemolytic even at a level of 0.70 mg/ml. Most amphiphiles, e.g. sodium deoxycholate, present in Fungizone, are haemolytic. Finally, AmB-loaded PEO-PBLA micelles can be freeze-dried and easily reconstituted in water. Afterwards, AmB is present in the intact PEO-PBLA micelles and remains nonhaemolytic. PMID- 9741921 TI - Biodegradable polyalkylcyanoacrylate nanoparticles for the delivery of oligonucleotides. AB - Antisense oligonucleotides with base sequences complementary to a specific RNA can, after binding to intracellular mRNA, selectively modulate the expression of a gene. However, these molecules are poorly stable in biological fluids and are characterized by a low intracellular penetration. In view of using oligonucleotides as active molecules, the development of polymeric particulate carriers was considered. Oligonucleotides were associated with biodegradable polyalkylcyanoacrylate nanoparticles through the formation of ion pairs between the negatively charged oligonucleotides and hydrophobic cations. Oligonucleotides bound to these nanoparticles were found to be protected from nuclease attack in cell culture media and their cellular uptake was increased as the result of the capture of nanoparticles by an endocytotic/phagocytotic pathway. The in vivo pharmacokinetic profile of oligonucleotides free or associated with nanoparticles has been investigated after intravenous administration to mice and the stability of these molecules has been evaluated by original methodology based on the use of polyacrylamide gel electrophoresis (PAGE) followed by multichannel radioactivity counting. Stability in vivo in the plasma and in the liver was shown to be improved when the oligonucleotides were adsorbed onto the nanoparticles. These results obtained both in vitro and in vivo open exciting perspectives for the specific delivery of oligonucleotides to the liver, thus considering this approach for the treatment of liver diseases (e.g. liver metastasis or hepatitis). PMID- 9741922 TI - 2-(Dimethylamino)ethyl methacrylate based (co)polymers as gene transfer agents. AB - Poly(2-(dimethylamino)ethyl methacrylate) (PDMAEMA) is a water-soluble cationic polymer, which is able to bind to DNA by electrostatic interactions. At a polymer/plasmid ratio above 2 (w/w) positively charged complexes were formed with a size around 0.2 microm. The transfection efficiency of polymer/plasmid complexes was evaluated in cell culture (COS-7 and OVCAR-3 cells) using a pCMV lacZ plasmid, encoding for beta-galactosidase, as a reporter gene. The optimal transfection efficiency was found at a PDMAEMA/plasmid ratio of 3-5 (w/w). Under these conditions 3-6% of the cells were actually transfected. Like other cationic polymers, PDMAEMA is slightly cytotoxic. This activity was partially masked by complexing the polymer with DNA. A pronounced effect of the molecular weight of the polymer on the transfection efficiency was observed. An increasing molecular weight resulted in an increasing number of transfected cells. Dynamic light scattering experiments showed that high molecular weight polymers (Mw>300 kDa) were able to condense DNA effectively (particle size 0.15-0.20 microm). In contrast, when plasmid was incubated with low molecular weight PDMAEMA, large complexes were formed (size 0.5-1.0 microm). Copolymers of DMAEMA with methyl methacrylate (MMA), ethoxytriethylene glycol methacrylate (triEGMA) or N-vinyl pyrrolidone (NVP) also acted as transfection agents. A copolymer with 20 mol % of MMA showed a reduced transfection efficiency and a substantial increased cytotoxicity compared with a homopolymer of the same molecular weight. A copolymer with triEGMA (48 mol %) showed both a reduced transfection efficiency and a reduced cytotoxicity, whereas a copolymer with NVP (54 mol %) showed an increased transfection efficiency and a decreased cytotoxicity as compared to a DMAEMA homopolymer. PMID- 9741923 TI - Effects of membrane-active agents in gene delivery. AB - A large variety of membrane-modifying agents have been used for the enhancement of DNA(lipo)polycation complex based gene transfer. The magnitude of improvement depends on the nature of the membrane-modifying agent and the (poly)cationic carrier. Within the lipid-free polymer-based systems (polyfection), ligand polylysine mediated gene transfer can be improved up to more than 1000-fold by pH specific endosomolytic peptides, glycerol, bacterial proteins or adenovirus particles. Ligand-polyethylenimine or dendrimer-based systems with per se higher efficiency are only slightly (about ten-fold) enhanced by endosomolytic agents. Membrane-active agents show only minor effects when applied to cationic lipid based gene transfer (lipofection) with DNA complexes formed under optimized conditions using an three- to four-fold excess of positive charges. Less positively charged lipofection complexes can be strongly improved by the addition of membrane-active peptides. PMID- 9741924 TI - A simple assay system for examination of the inhibitory potential in vivo of decoy RNAs, ribozymes and other drugs by measuring the Tat-mediated transcription of a fusion gene composed of the long terminal repeat of HIV-1 and a gene for luciferase. AB - Nucleic acid-based drugs, including antisense RNA and DNA, ribozymes and decoys appear to have potential for the suppression of the expression of specific genes. To allow the examination of the potential of such agents in vivo as anti-HIV drugs in standard laboratories, where facilities for handling live virions are not available, we constructed a simple assay system (HIV-1 model) that allows measurement of the extent of inhibition of Tat-mediated transcription of HIV-1 by nucleic acid-based drugs and other agents. In cells that harbor a stable chimeric long terminal repeat (LTR)-Luc construct (a fusion gene consisting of the LTR of HIV-1 and the gene for luciferase), total luciferase activity in an aliquot of cell lysate is dose- and promoter-dependent on transfection with a Tat expression plasmid, reflecting the character of the LTR promoter of HIV. When HeLa cells were co-transfected with the Tat expression plasmid and another plasmid that encoded the U6 promoter or the promoter of the gene for tRNA(Val) linked to the trans-activating response (TAR) sequence, total luciferase activity was inhibited by 60 or 40%, respectively. The inhibition was also dependent on the dose of the TAR expression plasmid. These results demonstrate the usefulness of this simple assay system for detection of the efficacy of a decoy RNA or a ribozyme in vivo, without a requirement for HIV-infected cells, by measurement of luciferase activity in vitro. PMID- 9741925 TI - A new non-viral DNA delivery vector: the terplex system. AB - A new DNA delivery vector (the terplex system) based on a balanced hydrophobicity and net surface charge between stearyl-poly(L-lysine), low density lipoprotein (LDL), and genetic material (i.e. plasmid DNA or antisense oligonucleotide) was developed. The pSV-beta-gal plasmid in terplex system showed a 2-5-fold increase in beta-galactosidase expression on murine smooth muscle cells (A7R5) compared to Lipofectin. Delivery of unmodified c-myb antisense oligonucleotide to A7R5 cells was also facilitated significantly by the terplex system, requiring as little as 5.4 nM of antisense oligonucleotide to achieve a 50% antiproliferative effect. Similar antiproliferative effect was observed when the c-myb antisense/terplex formulation was tested on CCD-32 Lu human lung fibroblasts. Characterization of the physical properties of the terplex system was performed using various techniques. Plasmid DNA was condensed by addition of stearyl-PLL and LDL, resulting in the terplex system of about 100 nm in diameter as shown by atomic force microscopy. A strong hydrophobic interaction between stearyl-poly(L-lysine) and LDL was registered by 1H-NMR spectrometry, showing a significant decrease in the epsilon-methylene signal of poly(L-lysine) backbone when stearyl-poly(L lysine) was mixed with LDL; however, this phenomenon was not observed with unmodified poly(L-lysine). Agarose gel electrophoresis revealed that electrophoretic mobility of the terplex system decreased with increasing amounts of stearyl-poly(L-lysine), indicating that the surface charge of the terplex system became more positive by addition of stearyl-poly(L-lysine). Zeta-potential measurement showed that the terplex system exerted a slightly positive charge (+2 mV) at a 1:1:1 weight ratio of plasmid DNA:LDL:stearyl-poly(L-lysine). The obtained results will be utilized in the design of more efficient and safer DNA delivery vectors for in vivo gene therapy. PMID- 9741926 TI - DNA-polycation nanospheres as non-viral gene delivery vehicles. AB - Nanospheres synthesized by salt-induced complex coacervation of cDNA and polycations such as gelatin and chitosan were evaluated as gene delivery vehicles. DNA-nanospheres in the size range of 200-750 nm could transfect a variety of cell lines. Although the transfection efficiency of the nanospheres was typically lower than that of lipofectamine and calcium phosphate controls in cell culture, the beta-gal expression in muscle of BALB/c mice was higher and more sustained than that achieved by naked DNA and lipofectamine complexes. This gene delivery system has several attractive features: (1) ligands can be conjugated to the nanosphere for targeting or stimulating receptor-mediated endocytosis; (2) lysosomolytic agents can be incorporated to reduce degradation of the DNA in the endosomal and lysosomal compartments; (3) other bioactive agents or multiple plasmids can be co-encapsulated; (4) bioavailability of the DNA can be improved because of protection from serum nuclease degradation by the polymeric matrix; (5) the nanosphere can be lyophilized for storage without loss of bioactivity. PMID- 9741927 TI - Nasal delivery of peptides: an in vitro cell culture model for the investigation of transport and metabolism in human nasal epithelium. AB - We investigated the transport- and metabolism properties of three peptides in monolayers of human nasal epithelial cells. The effective permeability coefficients of thyrotropin-releasing hormone, met-enkephalin and human recombinant insulin were found to be 4.5, 4.4 and 0.4 x 10(-7) cm/s, respectively. The permeability was inversely proportional to the molecular weight and one order of magnitude lower than in excised nasal mucosa of rabbits. The metabolic cleavage of thyrotropin-releasing hormone (TRH) to the free acid by cytosolic prolyl-endopeptidase was also detected in human nasal cell monolayers, suggesting that ca. 10% of the total amount of TRH is transported via a transcellular pathway. Met-enkephalin is a substrate for aminopeptidases, located on the apical membrane of nasal epithelial cells. Metabolites and enzyme activity are comparable with literature data. Our studies demonstrate that not only morphological, but also functional properties of human nasal epithelial cells are preserved under in vitro conditions. Such a cell culture model based on human nasal cells could be beneficial for the characterization of peptide transport on a cellular level and for investigation of the absorption enhancer mechanism. Further studies are necessary, however, to establish correlations between in vitro permeabilities in cell cultures and nasal drug absorption in animals and humans. PMID- 9741928 TI - Biological response to a synthetic cornea. AB - We have designed a synthetic cornea that has a transparent hydrogel optic and a porous skirt. The device has been implanted in rabbit corneas. We have shown that keratocytes migrate into the device and deposit a complex extracellular matrix. The immediate response is detected in the surrounding stroma, and the secondary response is seen after cells have deposited a matrix in the disc. After implantation, a decrease in keratan sulfate accompanied by an increase in dermatan sulfate was detected in the surrounding tissue compared to the unwounded corneal stroma. The glycosaminoglycans in the disc resemble that of an injured stroma. The appearance of heparan sulfate and growth factors, bFGF and TGFbeta, was not detected until 6 weeks after implantation. The growth factors were detected at the interface between the device and the tissue and become more diffuse over time. Methods of controlled release in vivo were used to enhance the rate of fibroplasia and wound repair. While these were successful in the cornea itself, when combined with the synthetic cornea the response was magnified and the initial attempts yielded neovascularization and edema. Currently, efforts are being directed at controlling the release within the porous haptic so that fibroplasia is enhanced while minimizing an inflammatory response. PMID- 9741929 TI - Novel mucosal immunization with polysaccharide-protein conjugates entrapped in alginate microspheres. AB - A novel mucosal immunization was examined using biocompatible and biodegradable alginate microspheres containing a conjugate of polysaccharide antigen and cholera toxin B subunit (CTB). In order to prepare the alginate microspheres with diameters of less than 5 microm, a new diffusion-controlled interfacial gelation technique was developed. Also, in order to improve the mucosal immune response, a pneumococcal capsular polysaccharide type 19 (PS19) was conjugated to the CTB (PS19-CTB). This conjugate was subsequently encapsulated into the alginate microspheres. The loading content of PS19-CTB to the alginate microspheres was 60%. An in vitro sustained release pattern was observed with the antigen-loaded microspheres, showing 80% antigen release within one day. Mucosal and systemic immunities following oral immunization with the alginate microspheres were studied. Balb/c mice were immunized perorally three times at intervals of two weeks. Peroral immunization with 25 microg of PS19-CTB entrapped in the alginate microspheres evoked both the mucosal IgA and systemic IgM responses to PS19 in small intestine and in sera, respectively. The results suggest that both the mucosal and systemic antibody responses could be induced by oral administration of the PS19-CTB antigen entrapped in alginate microspheres. PMID- 9741930 TI - Cross-linked high amylose starch for controlled release of drugs: recent advances. AB - Cross-linked high amylose starches have been developed as excipients for the formulation of controlled-release solid dosage forms for the oral delivery of drugs. Advantages of this new class of excipients include cost-effectiveness, readily accessible industrial manufacturing technology, high active ingredient core loading and the possibility of achieving a quasi zero-order release for most drugs. In addition to the latter, other features distinguish cross-linked high amylose starches from other excipients used to prepare hydrophilic matrices. Among these are the absence of erosion, the limited swelling and the fact that increasing cross-linking degrees results in increased water uptake rate, drug release rate and equilibrium swelling. Thus the goal of the present study was to gain some insights into the mechanism of drug release control by matrices of cross-linked high amylose starch. Water transport kinetics and dimensional changes were studied in matrices placed in water at 37 degrees C by an image analysis technique. The results show that in the first 5 min, a gel layer is formed at the surface of the tablet, after which the gel front seems to halt its progression toward the center of the tablet. Water continues to diffuse through the front and to invade the core. As a consequence, this latter swells, with a predominance for radial swelling. Equilibrium swelling is reached over 3 days, when the water concentration in the tablet becomes homogeneous and the whole tablet gelifies. Solid-state 13C-NMR were acquired on cross-linked high amylose starch powders, tablets and hydrated tablets with varying cross-linking degrees. They show a predominance of the V-type single helix arrangement of amylose in the dry state irrespective of the cross-linking degree. Upon hydration, the homologues with a low cross-linking degrees show a transition from the V to the B type double helix arrangement. It is therefore hypothesized that the capacity of amylose to undergo the V to B transition is an important factor in controlling water transport and drug release rate. Finally applications to different drugs are reviewed briefly. They illustrate the versatility of this technology as generic versions of zero order OROS drug (Efidac) and Fickian release conventional matrices (Voltaren SR) were developed and successfully tested in pilot clinical studies to be bioequivalent to the references. These studies further showed that cross-linked high amylose starch matrices have the lowest inter-subject variability among the systems tested and show a total absence of food effect. PMID- 9741931 TI - Controlled delivery of antibacterial proteins from biodegradable matrices. AB - Prosthetic valve endocarditis is an infrequent, but serious complication of cardiac valve replacement. The infection is caused by the adherence of bacteria to the prosthetic valve or to tissue at the site of implantation. Recently it was shown that antibacterial peptides from blood platelets are involved in clearance and killing of bacteria adhering to vegetations induced in a model for prosthetic valve endocarditis using rabbits. The application of these antibacterial proteins in a release system, incorporated in the Dacron sewing ring of the prosthetic heart valve would diminish the incidence of endocarditis. In this study a release system for small cationic proteins based on cross-linked gelatin was developed and characterised. Furthermore, the system was evaluated with respect to the uptake and in vitro release of lysozyme, a small cationic protein that was chosen as a model protein for small cationic antibacterial proteins. Variation of gelatin type (A and B), and cross-link density resulted in differences in swelling, thermal behaviour, and number of charged groups. Lysozyme uptake was proportional to swelling, but was governed by the number of anionic groups. The latter was also observed for the release profiles: when the amount of free carboxylic acids is higher (gelatin B compared to gelatin A), the lysozyme release lasts for a longer time period. The release into solidified agarose medium, as a model for heart muscle tissue, was measured. After 50 h, 40-100% of the lysozyme was released, which is in accordance with the aimed release period of 24-48 h. The adsorption experiments in vitro suggest an influence of the electrostatic interactions between lysozyme and gelatin. This hypothesis was validated with a mathematical model which takes both diffusion and adsorption interactions into account. PMID- 9741932 TI - Extracellular matrix for a rechargeable cell delivery system. AB - Above a critical concentration, aqueous polymer solutions of N isopropylacrylamide copolymers with small amounts of acrylic acid, synthesized in benzene by radical polymerization, exhibited four distinct phases as the temperature increased; clear solution, opaque solution, gel and shrunken gel. The transition between the opaque solution phase and the gel phase was in the range of 30-34 degrees C and was reversible without syneresis and noticeable hysteresis under the experimental conditions used in this study. Islets of Langerhans, isolated from Sprague-Dawley rat pancreata and entrapped in the gel matrix, remained viable, with no significant decrease in insulin secretion function in vitro for one month. When islets were encapsulated with the gel matrix in hollow fibers [molecular weight cut-off (MWCO)= approximately 400000] and were exposed to dynamic changes in glucose and theophylline concentrations, their insulin secretion patterns demonstrated a smaller lag time and higher amplitude in insulin release than islets entrapped in a conventional alginate matrix under the same experimental conditions. From these two observations, i.e. gel reversibility and islet functionality in the matrix observed in in vitro experiments, the N isopropylacrylamide copolymers with acrylic acid synthesized in this study are optimum candidates for the extracellular matrix in a diffusion chamber-type cell delivery system in order to recharge the entrapped cells when cell functionality in the system decreases. PMID- 9741933 TI - Partially unfolded proteins efficiently penetrate cell membranes--implications for oral drug delivery. AB - We have previously reported on the biological activity of members of a library of low molecular weight compounds (carriers) that enable the oral delivery of proteins (Milstein, Proceedings of the 1995 Miami Bio/Technology Winter Symposium on Protein Engineering and Structural Biology, IRL Press at Oxford University Press, 1995, p. 13; Leone-Bay et al., J. Med. Chem. 38 (1995) 4263-4269; Leone Bay et al., J. Med. Chem. 39 (1996) 2571-2578; [1-3]). When rats or primates are orally administered a solution of carrier and either recombinant human alpha interferon (rhIFN), insulin or recombinant human growth hormone (rhGH) significant serum concentrations of the proteins are detectable. The transport activity of these compounds is positively correlated with their structural effects on the protein molecules. Direct measurement of the interaction of these carrier molecules with the proteins indicates that they reversibly destabilize the native state of the molecule favoring a partially unfolded conformation. Apparently these intermediate protein conformations are transport competent and are able to be absorbed through the intestinal tissue and into the bloodstream. Since the measured binding of the carriers to the partially unfolded proteins is relatively weak (Kb = 100 M(-1)) and the systemic activity of the proteins appears to be unaffected, the changes in the structure of the proteins are manifestly reversible. PMID- 9741934 TI - Inhalation delivery systems with compliance and disease management capabilities. AB - Non-compliance with prescribed medication is a major reason for poor therapeutic outcomes, leading to unnecessary contributions to healthcare costs. Poor technique in self-administration of inhalation therapy is a special type of non compliance associated with this route of administration. However, pulmonary drug delivery has fundamental advantages for therapy of diseases of the respiratory tract because it is site-directed. The lung is also a promising portal for drug delivery into the systemic circulation. Incorporation of microprocessors into pulmonary drug delivery systems facilitates sophisticated compliance management of chronic diseases such as asthma and diabetes. Microprocessor-assisted systems afford control of patients' administration technique during the therapeutic inhalation event, thus leading to efficient and reproducible regional deposition of the inhaled drug or diagnostic agent. SmartMist is a hand-held asthma disease management device that aids patients to use optimally metered dose inhalers. It also measures pulmonary lung function and provides a long term downloadable electronic record of the therapeutic and diagnostic events. The AERx pulmonary delivery system utilizes similar microprocessor capabilities; however, it employs a novel means of generating aqueous aerosols from unit dose packages, thus providing a broad inhalation technology base for delivery of a wide variety of therapeutic and diagnostic agents into the respiratory tract, and via the lung into the systemic circulation. PMID- 9741935 TI - Development of liposomal anthracyclines: from basics to clinical applications. AB - The pharmacokinetics of liposome-encapsulated drugs are controlled by the interplay of two variables: the rate of plasma clearance of the liposome carrier, and the stability of the liposome-drug association in the blood stream. The pharmacokinetic properties of the liposomal drug, the vesicle size of the liposome carrier and the vascular permeability of individual tissues will determine the extravasation and biodistribution profile. The pharmacokinetics of polyethylene-glycol-(PEG)-liposomal doxorubicin are characterized by an extremely long circulating half-life, slow plasma clearance and reduced volume of distribution compared to free doxorubicin. These carrier systems show an improved extravasation profile with enhanced localization in tumors and superior therapeutic efficacy in comparison to doxorubicin in free form. These properties are the result of an optimized liposome composition and of a special drug-loading method which produces stable and long-circulating carriers. In clinical studies, doxorubicin encapsulated in PEG-coated liposomes shows a unique pharmacokinetic toxicity profile and promising antitumor activity. PMID- 9741936 TI - Boundary layer drug delivery using a helical catheter. AB - BACKGROUND: A catheter-based approach for local endovascular drug delivery has been developed. The catheter is deployed percutaneously, while the end of the catheter is in the form of a helix that is placed just proximal to the vascular site to be treated. The helices are in contact with the vessel wall. A number of small holes is drilled in the coils of the catheter through which drug is infused, so that the infused drug remains within the blood fluid 'boundary layer' adjacent to the vessel wall. This approach is expected to be highly efficient for administration of antithrombotic and antiproliferative agents that target processes leading to vascular occlusion, heart attacks, and strokes. METHODS: The helical catheter was qualitatively evaluated using optical dye density measurements of Evans blue dye infused using an in vitro steady flow system under a physiologic range of conditions. To further demonstrate the efficiency of the technique, its capacity to inhibit thrombosis was evaluated in a baboon thrombosis model. The catheter was inserted into a femoral arteriovenous shunt (blood flow rate = 100 ml/min) and placed proximal to a segment of highly thrombogenic Dacron vascular graft (4.0 mm i.d.). Integrelin (an inhibitor of platelet glycoprotein IIb/IIIa; doses: 0.25-1.0 microg/min) and hirudin (an antithrombin; doses: 10-100 microg/min) were used to inhibit thrombus formation. RESULTS: Experimental flow visualization studies demonstrated that high concentrations of the infused Evans blue dye were retained near the vessel wall. In the animal experiments, platelet deposition on the Dacron graft surface was reduced by 82-97% (Integrelin) and 68-92% (hirudin) over 1-2 h of blood exposure. The local antithrombotic effects produced were found to be 200-fold and 30-fold more efficient than systemic administration of the same agents. CONCLUSIONS: Local drug infusion using the helical catheter approach can achieve high drug concentration levels at target sites, may avoid systemic effects, and can reduce cost of therapy by reducing total drug requirements. PMID- 9741937 TI - Polycation-DNA complexes for gene delivery: a comparison of the biopharmaceutical properties of cationic polypeptides and cationic lipids. AB - DNA plasmids formed particulate complexes with a variety of cationic polyamino acids and cationic lipids, which were used to transfect mammalian cells in culture. Complexation was studied by assaying for exclusion of ethidium using a fluorometric assay, which indicated that complexation with cationic polyamino acids took place with utilisation of the majority of charged functional groups. The particle sizes and zeta potentials of a range of complexes were determined. Generally polyamino acids formed uniform particles 80-120 nm in diameter in water, but their particle size increased on dilution of the particles in electrolytes or cell culture media. The efficiency of transfection was compared using complexes of pRSVlacZ, a reporter construct which expressed beta galactosidase under the control of the Rous sarcoma virus promoter. Positively charged DNA/polyamino acid complexes were taken up by cells but required an endosomolytic agent, such as chloroquine, to facilitate transfection. Polyornithine complexes resulted in the highest levels of expression, in comparison with other homopolyamino acids (polyornithine>poly-L-lysine=poly-D lysine>polyarginine). Copolyamino acids of lysine and alanine condensed DNA but were less active in transfection experiments. Copoly(L-Lys, L-Ala 1:1) was inactive even in the presence of chloroquine. In contrast DNA/cationic lipid complexes transfected cells spontaneously, and chloroquine did not improve the extent of expression, rather it usually reduced efficiency. There was little correlation between comparative efficiencies of lipid complexes between cell lines suggesting that the nature of the cell membrane and differences in mechanisms of internalisation were determinants of efficiency. In an effort to explore better cell culture models for gene delivery, monolayers of Caco-2 cells were transfected in filter culture. As the cells differentiated and formed a polarized monolayer, expression of beta-galactosidase was reduced until at day 27 expression was not significantly different from basal activity. The Caco-2 filter culture model merits further attention as a model of gene delivery to epithelial surfaces, such as would be encountered in the lung after inhalation. PMID- 9741938 TI - Targeted delivery of plasmid DNA complexed with galactosylated poly(L-lysine). AB - Galactose was introduced to poly(L-lysine) (PLL) with an average molecular weight of 13,000 to develop a hepatocyte-specific carrier for gene drugs. The pharmacokinetic characteristics of a model plasmid, pCAT (plasmid DNA encoding chloramphenicol acetyltransferase reporter gene), complexed with galactosylated PLL (Gal-PLL) was studied in mice in relation to its physicochemical properties. pCAT/Gal-PLL complex at a ratio of 1:0.6 (w/w) has a zeta potential of -20 mV and a mean particle size of about 180 nm. After intravenous injection, [32P]pCAT/Gal PLL was rapidly eliminated from the circulation and preferentially taken up by the liver's parenchymal cells. The hepatic uptake of [32P]pCAT/Gal-PLL was significantly inhibited by prior administration of Gal-bovine serum albumin, suggesting that the uptake was mediated by the asialoglycoprotein receptors on hepatocytes. In vitro transfection experiments using a hepatoma cell line expressing the asialoglycoprotein receptor revealed that pCAT/Gal-PLL gave a high CAT gene expression whereas pCAT complexed with unmodified PLL failed to transfect the cells. PMID- 9741939 TI - Flupenthixol treatment for cocaine abusers with schizophrenia: a pilot study. AB - Cocaine use is common among individuals with schizophrenia and schizoaffective illness, with a prevalence ranging from 15-60% of patient samples. It is hypothesized that some schizophrenic cocaine abusers may use cocaine as an attempt to improve anhedonic symptoms or combat neuroleptic side-effects. Flupenthixol (FLX) has the distinct advantage of being both a neuroleptic medication and a potential treatment for cocaine abuse. We evaluated the efficacy of FLX in this dually diagnosed population in an open pilot study consisting of a 4-week inpatient phase and a 6-week outpatient phase. Eight individuals were initially cross-tapered off their neuroleptic medication and were given FLX in a dose of 40 mg of the decanoate every 2 weeks. Psychiatric symptomatology was assessed weekly, using the Positive and Negative Symptom Scale (PANSS) and the Beck Depression Inventory (BDI). Medication side-effects were monitored weekly, using the Simpson Neurological Rating Scale and the Abnormal Involuntary Movement Scale (AIMS). Substantial improvement in psychiatric symptomatology was noted when preadmission scores were compared to scores obtained during the last week of study enrollment. On the PANSS, positive symptom scores and negative symptom scores decreased by 31% and 29%, respectively. Similarly, BDI scores decreased by 57%. Comparing preadmission urine results to those for the last 6 weeks of enrollment in the study showed that cocaine-positive urines decreased by 28%, although most of the patients had a reduction of >75%. Missed clinic visits decreased by 26%. Thus, FLX was well-tolerated by schizophrenic cocaine abusers, suggesting that FLX may be useful for the treatment of this dually diagnosed population. PMID- 9741940 TI - Antisocial tendency among drug-addicted adults: potential long-term effects of parental absence, support, and conflict during childhood. AB - This study examined the relationship between perceptions of parent-child relations in the family of origin and antisocial tendency in a sample of drug addicted adults. Data included retrospective accounts of childhood family factors, adolescent antisocial tendency, and self-reported hostility and risk taking prior to treatment entry. A developmental model was tested that included adolescent antisocial tendency as a mediator of the relationship between childhood parenting factors and adulthood antisocial tendency. The effects of parental support and conflict were found to operate primarily through adolescent measures. Specifically, lower levels of parental support and higher levels of conflict with parents predicted greater adolescent antisocial tendency, which in turn predicted more hostility and risk-taking in adulthood. Thus, parental support appears to serve as a buffer against deviant behavior and drug use. PMID- 9741941 TI - Correlates of HIV seropositivity and HIV testing among out-of-treatment drug users. AB - This study: 1) examined the rate and correlates of human immunodeficiency virus (HIV) seropositivity; and 2) assessed whether self-selection in HIV testing influenced the rate and correlates of HIV seropositivity in a group of out-of treatment drug users. Data were collected from 856 out-of-treatment drug users in Philadelphia between January 1993 and August 1994. Seventy-four percent of the sample elected to take an HIV test that was included in the project in which the drug users were enrolled, and of these, 11% were HIV positive. Multivariate analyses indicated that those who were younger, those who had an injection-drug using sex partner, and those who reported no recent sexual activity were more likely to be HIV positive. An examination of the multivariate correlates of HIV testing indicated that subjects who took an HIV test had higher rates of participation in some risk behaviors than did subjects who did not take an HIV test, but lower rates for other risk behaviors. None of the correlates of HIV seropositivity were correlates of HIV testing. Although the generalizability of the HIV seroprevalence rate is unclear, it is probable that the correlates of HIV seropositivity are generalizable to the total sample. The results of this study indicate the importance of interventions that target sexual risk behavior among out-of-treatment drug users, and of assessing the impact of self-selection bias whenever the rate and correlates of HIV seropositivity are examined. PMID- 9741942 TI - Substance-use situations and abstinence predictions in substance abusers with and without personality disorders. AB - Rates of personality disorders (PDs) in substance abusers are higher than in the general population. Comorbid PDs are believed to complicate the treatment of addicted patients: in addition to having more severe substance-use disorders and life problems, personality-disordered patients may use substances differently than their peers without Axis II diagnoses. In a sample of 339 adults receiving inpatient treatment for alcohol or drug abuse/dependence, 71.7% received Axis II diagnoses, and they presented a more severe clinical picture. They also had more self-reported "impulsive" substance use and use of drugs or alcohol in positive situations. Different groups of personality-disordered patients had different patterns of self-efficacy for abstinence for hypothetical future situations. PMID- 9741943 TI - Illicit drug use by women with disabilities. AB - As in the general female population, women with disabilities face a number of situations that encourage illicit drug use, such as low self-esteem, peer pressure, and family history of substance abuse. In addition, women with disabilities more frequently encounter problems of personal adjustment, unusual developmental experiences, easy access to prescription drugs, unemployment, and medical or health-related difficulties. Using a random sample of 900 women with disabilities, we conducted a study of the patterns of illicit drug use and risk factors relating to illicit drug use among women with various disabilities. Multiple regression analyses revealed that age, illicit drug use by a best friend, being a victim of substance-abuse-related violence, and attitudes toward substance use by people with disabilities were significantly related to illicit drug use by the study population. The implications of these findings are discussed. PMID- 9741944 TI - Women, violence with intimates, and substance abuse: relevant theory, empirical findings, and recommendations for future research. AB - Evidence from the disparate domains of anthropology, criminology, psychology, and sociology indicates that women are involved in many of the same acts of aggression and violence as men, and that substance use may play an important role in these acts. Yet little is known of the pathways between violence and drugs for women. The aims of this paper are threefold: 1) To review and integrate existing literature addressing female violence and substance abuse, presenting available epidemiology, theories, and research applicable to the study of this problem. 2) To examine the political and methodological obstacles to conducting systematic research on female aggressiveness. 3) To offer recommendations for future epidemiological, preventive, and therapeutic research efforts in this vital yet grossly understudied area. PMID- 9741945 TI - Drug abuse treatment and risky sex: evidence for a cumulative treatment effect? AB - This paper presents evidence regarding the possibility of a cumulative effect of drug-abuse treatment on reducing risky sexual behavior among individuals entering drug-abuse treatment in the United States from 1991-1993 and participating in the Drug Abuse Treatment Outcome Study (DATOS). Analyses were done of the relationship between lifetime treatment exposure and risky sex by drug users during the year before intake into DATOS. Analyses controlled for age, drug-use severity, criminal history, antisocial conduct disorder, and other factors that might have confounded the relationship between treatment exposure and risky sex. Results indicated that users with more lifetime treatment exposure had lower scores for risky sex. This finding is consistent with the hypothesis that successive episodes of treatment may have long-term cumulative effects on drug users' HIV-risk behavior. PMID- 9741946 TI - Anxiety and alcohol abuse in patients in treatment for depression. AB - Anxiety may be an important factor in explaining the prevalence of alcohol abuse among depressive patients. However, it is unclear whether anxiety has effects that are independent of other core symptoms in depression, and whether it is linked to alcoholic problems in both sexes. The present study of hospitalized depressive patients found a strong association between anxiety and alcohol abuse for women, and a weaker association for men. These effects were independent of severity of depression and global pathology. Whereas the correlation appeared to be linear for men, with each higher level of anxiety being associated with more alcohol problems, for women depressives those in a moderate-anxiety subgroup had the most difficulty with alcohol. PMID- 9741947 TI - The effects of drinking on health of older adults. AB - As part of a larger health-promotion project focused on safer use of alcohol and medications by older people, 826 persons living in a small community in Eastern Ontario participated in a survey conducted by nurses in the participant's home. The survey interview varied in length from 30 to 210 minutes and covered alcohol use, medication use, health status, and other aspects of life. This report examines the relationship between drinking practices and self-reported health (overall rating of health, hospital admissions, etc.) as identified by the survey. Among survey participants who abstained from alcohol during the 12 months prior to the survey, former drinkers reported significantly poorer health than did lifetime abstainers and previous drinkers who happened to abstain in the previous year but considered themselves to be infrequent drinkers rather than former drinkers. Among current drinkers, poorer health was associated with drinking more on each occasion of drinking and with a greater total overall volume of alcoholic beverages consumed (as estimated from the drinker's usual weekly consumption), but not with more frequent drinking. Even when controlling for sex, age, education, and depressant medication use, quantity of alcoholic beverage consumption per occasion and overall volume consumed were found to contribute significantly to predicting perceived health. The implications of the findings are discussed in terms of the best ways to assess the risk level of alcohol use among older people, and with recommendations for safer alcohol use. PMID- 9741948 TI - Gender, health beliefs, health behaviors, and alcohol consumption. AB - We conducted a study of the relationship between health beliefs, health practices, and alcohol consumption among women and men using the Health Belief Model (HBM). The study sample (N = 41,104) was drawn from the 1990 National Health Interview Survey. A 15% random sample was selected from the total data set for the purpose of selecting variables. Alcohol consumption was defined as a health-related behavior and was regressed on demographic characteristics, health beliefs, and health practices. Quantity and frequency of alcohol consumption were assessed for each gender, using the HBM. The HBM explained similar amounts of the variance in alcohol use for women and men. Quantity of alcohol consumed accounted for more variance than did frequency of consumption. We conclude that health beliefs and behaviors are related to alcohol consumption after adjustment for demographic characteristics, and that differences exist between perceived risks and behaviors for both women and men. PMID- 9741950 TI - Irritable bowel syndrome (IBS) and alcohol abuse or dependence. AB - Irritable bowel syndrome (IBS) has been reported in 10-22% of adults. Using a semistructured clinical interview to study the prevalence of IBS, we compared 31 patients seeking treatment for alcohol abuse or dependence in an outpatient setting with an age- and sex-matched control group of 40 patients who were seeking treatment in a general physician's office for other medical illnesses. The control group did not have any Axis I disorders. IBS was diagnosed according to the criteria of Drossman et al. Thirteen (41.9%) patients with alcohol abuse or dependence met the criteria for IBS, in contrast to 1 (2.5%) patient in the control group. We conclude that IBS is common and frequently underdiagnosed in patients with alcohol abuse or dependence. PMID- 9741949 TI - Isometric muscle strength in alcoholic and nonalcoholic liver-transplantation candidates. AB - Intensity of isometric muscle contractions was measured in alcoholic subjects with cirrhosis (N = 42), nonalcoholic subjects with cirrhosis (N = 33), and normal controls (N = 31). Muscle strength and endurance were comparable in the alcoholic and nonalcoholic cirrhotic subjects for all variables. Both cirrhotic groups were inferior to normal controls for all variables. The quantity x frequency (Q x F) index reported for the period during peak alcohol consumption correlated with 6 muscle-force variables, accounting for 9-20% of the variance. Alcoholic and nonalcoholic cirrhotic subjects did not differ in Quality of Life Inventory (QOLI) scales. Significant correlations, however, were found for the alcoholic cirrhotic subjects but not for the nonalcoholic cirrhotic subjects between quality-of-life indices and muscle strength and endurance. Muscle weakness is thus differentially associated with quality of life in alcoholic cirrhotic subjects as compared with nonalcoholic subjects with cirrhosis even though level of strength and endurance in the two groups is comparable. PMID- 9741951 TI - Metabolic fate of S-(-)-pulegone in rat. AB - 1. S-(-)-pulegone was administered orally to rat (250 mg/kg) and the nature of the urinary metabolites was investigated. Eleven metabolites, namely S-(-) menthofuran, piperitone, piperitenone, p-cresol, 5-hydroxypulegone, 4 methylcyclohexenone, 3-methylcyclohexanone, isopulegone, pulegol, 7 hydroxypiperitone and benzoic acid, have been isolated from rat urine. It is assumed that menthofuran, isopulegone and 4-methylcyclohexenone retain the stereochemistry of the parent compound, whereas in other metabolites the stereochemistry at the asymmetric centres is not known. 2. The relative amounts of various major metabolites present in the total urine extracts from the R-(+) and S-(-)-pulegone-treated rat were established by glc analyses. Urine samples of rats treated with R-(+)-pulegone contained higher levels of p-cresol and piperitenone than in similar experiment carried out with S-(-)-pulegone, whereas the levels of unmetabolized pulegone, piperitone and benzoic acid were considerably higher in the urine of rat treated with S-(-)-pulegone than in a corresponding experiment with R-(+)-pulegone. 3. Phenobarbital-induced rat liver microsomes converted S-(-)-pulegone to S-(-)-menthofuran (VII) and piperitenone (III) in the presence of NADPH and O2. The levels of VII and III were significantly higher in similar experiments carried out with R-(+)-pulegone. 4. Based on these studies, metabolic pathways for the biotransformation of S-(-) pulegone in rat have been proposed and possible reasons for the observed difference in the toxicity mediated by these two enantiomers are discussed. PMID- 9741952 TI - Inhibition of murine hepatic cytochrome P450 activities by natural and synthetic phenolic compounds. AB - 1. The effect of the phenolic compounds protocatechuic acid, chlorogenic acid, tannic acid, gallates and silybin on ethoxyresorufin O-dealkylase (CYP1A1), methoxyresorufin O-dealkylase (CYP1A2) and pentoxy-O-dealkylase (CYP2B) was examined in mouse liver microsomes from induced animals. 2. All compounds tested could inhibit cytochrome P450-mediated enzyme activities, but to different extents. Tannic acid was the most potent inhibitor, especially toward EROD activity with an IC50=2.6 microM. Synthetic dodecyl gallate was also relatively selective toward this enzyme activity with an IC50=120 microM. 3. Protocatechuic acid, chlorogenic and silybin were more selective towards PROD and MROD activities. Their relative inhibitory potency for PROD activity was as follows: chlorogenic acid > protocatechuic acid > silybin > dodecyl gallate > propyl gallate. Protocatechuic acid was a more effective inhibitor of MROD activity than chlorogenic acid, and propyl gallate more effective than dodecyl gallate. Thus, no clear structure-activity or selectivity relationship was observed. 4. Analysis of the kinetics of inhibition revealed that the inhibition in most cases was non competitive in nature. PMID- 9741953 TI - Characterization of biliary metabolites of 4-n-nonylphenol in rainbow trout (Oncorhynchus mykiss). AB - 1. [R-2,6-3H]-4-n-nonylphenol was synthesized and a single dose (5 mg, 1850 KBq) orally administered to rainbow trout. After 48 h, the radioactivity present in the bile amounted 5.5%. More than ten biliary metabolites were separated by hplc and collected for subsequent mass spectrometry analysis. The metabolic profile was totally modified by beta-glucuronidase hydrolysis, showing that most of the metabolites were glucuronic acid conjugates. 2. Conjugated metabolites were identified by lc-ms analysis and their aglycones were analysed by gc-ms analysis as TMS and acetyl derivatives. 3. The major metabolite accounted for 52+/-11% of the biliary radioactivity and was identified as nonylphenol-glucuronide. 4. Nonylphenol was hydroxylated at both omega and omega-1 positions of the alkyl chain, giving 9-hydroxynonylphenol and 8-hydroxynonylphenol. 5. 9 Hydroxynonylphenol was oxidized to the corresponding acid, and subsequently beta oxidized, yielding 7-(4-hydroxyphenyl)heptanoic acid, 5-(4 hydroxyphenyl)pentanoic acid, 3-(4-hydroxyphenyl)propionic acid and 3-(4 hydroxyphenyl)-2-propenoic acid. PMID- 9741954 TI - Effect of a new rifamycin derivative, rifalazil, on liver microsomal enzyme induction in rat and dog. AB - 1. The effect of a new rifamycin derivative, rifalazil (KRM-1648), on liver microsomal enzyme induction was studied in rat and dog with repeated oral administration of the compound. Relative liver weight, cytochrome b5 and P450 contents, enzyme activities of NADPH-cytochrome c reductase, aniline hydroxylase, p-nitroanisole O-demethylase, aminopyrine N-demethylase, and erythromycin N demethylase were measured. 2. In rat, rifalazil treatment at 300 mg/kg/day for 10 days increased cytochrome b5 content but it did not affect liver weight, P450 content or enzyme activities. In contrast, rifampicin and rifabutin increased relative liver weights, cytochrome contents and enzyme activities under similar conditions. 3. In dog, rifalazil did not affect any parameters at 30 or 300 mg/kg/day for 13 weeks. 4. These findings indicate that rifalazil is not an enzyme inducer in rat and dog. This property differs from other rifamycin derivatives such as rifampicin and rifabutin. PMID- 9741955 TI - Disposition of butanal oxime in rat following oral, intravenous and dermal administration. AB - 1. The disposition of [1-14C]butanal oxime (BOX) was determined in the rat after oral, i.v. and dermal administration. 2. Oral doses of [14C]BOX (2 and 20 mg/kg) were predominantly excreted in the urine (> 42%) and converted to 14CO2 (> 30%) and about 10% of the dose remained in the tissues 72 h post-dosing. 3. Eight and 16% of a 2 and 20 mg/kg dermal dose of BOX, respectively, were absorbed, due in part to rapid volatilization from the surface of the skin. 4. Oral doses of BOX were transformed into several polar and/or anionic metabolites that include sulphate conjugates and a significant amount of thiocyanate. 5. The effect of inhibitors on the metabolism of BOX was investigated using 1-aminobenzotriazole (ABT; an inhibitor of diverse cytochrome P450s) and trans-1,2-dichloroethylene (DCE; an inhibitor of CYP2E1). No thiocyanate anion was detected in the urine of rat treated with DCE or ABT. ABT markedly increased the production of 14CO2 and excretion as volatile metabolites. DCE had no effect on 14CO2 excretion, but increased exhalation of radiolabel. ABT also effectively blocked the expression of toxic effects attributable to cyanide in rat given near-lethal doses of BOX. 6. The data are consistent with two distinct pathways of metabolism for BOX, (1) reduction to an imine, hydrolysis and subsequent conversion of butyraldehyde to 14CO2 and (2) CYP3A-catalysed dehydration of BOX to butyronitrile followed by CYP2E1-catalysed release of cyanide. PMID- 9741956 TI - Differential in vitro hepatic and intestinal metabolism of ifosfamide in the rat. AB - 1. Metabolism of ifosfamide (IF) in intestinal and hepatic microsomes has been investigated in the rat. The generation of three primary metabolites, 4 hydroxyifosfamide (HOIF), N2-dechloroethylifosfamide (N2D) and N3 dechloroethylifosfamide (N3D), was followed. 2. Microflora in rat small intestine showed no metabolic activity towards IF. The overall metabolic activity was higher in the hepatic microsomes than in the intestinal microsomes, and the hydroxylation pathway accounted for 53.6% of total IF metabolism in the hepatic microsomes. In contrast, hydroxylation of IF in intestinal microsomes was only 9.8% of the total monitored metabolic activity and N3-dechloroethylation of IF was a major pathway, constituting 73.0% of the monitored activity. 3. In summary, the intestinal metabolism of IF was demonstrated for the first time and these in vitro data indicate that the intestinal metabolism of IF could contribute significantly to the overall first-pass metabolism. PMID- 9741957 TI - In vivo metabolism of a novel cholecystokinin B (CCK-B) antagonist in rat and dog plasma and rat faeces. AB - 1. The in vivo metabolism of a novel CCK-B antagonist ((+)-N-[1-(adamantane-1 methyl)-2,4-dioxo-5-phenyl-2,3,4,5-tetrahydro-1H -1,5-benzodiazepin-3-yl]-N' phenylurea, GV150013X) was investigated using rat and dog plasma (male and female) and rat faeces samples after administration of GV150013X. 2. Four monohydroxy and four dihydroxy metabolites of GV150013X were identified by comparison with authentic standards using hplc and results from previous in vitro studies. 3. In both rat and dog plasma, GV150013X was converted to one major and other minor metabolites. 4. Qualitatively there is no species or sex differences in the formation of metabolites except that minor metabolite M1 was not detected in dog plasma. 5. Traces of GV150013X and the major metabolite were seen in rat plasma sample 24 h after administration. 6. Hplc with UV and radiochemical detection was used to identify metabolites. Major, non-labelled GV150013X metabolites from rat faeces were collected for characterization by nmr. PMID- 9741958 TI - Changes in the enzymatic activities of beagle liver during maturation as assessed both in vitro and in vivo. AB - 1. We have examined changes in caffeine and trimethadione (TMO) metabolism in vivo, agents which are used as probe drugs. In this study the total body clearance (Cl) of caffeine and TMO was low 1 week after birth (week 1), increased rapidly from week 3, peaked and then decreased gradually until reaching the level for the mature, adult dog. The elimination half-life (t1/2) of caffeine and TMO was prolonged during week 1; however, it then gradually became shorter. Gradually it became longer and reached the level for the adult dog. The apparent volume of distribution (Vd) of caffeine did not change throughout the study. However, the Vd of TMO was only high during week 1. 2. The in vitro changes in a variety of typical substrates for seven different cytochrome P450 (CYP) isozymes were examined. In this study three different patterns of metabolism can be identified: (1) activity is low immediately after birth, increases, peaks and then decreases to the adult dog level (p-nitroanisole; CYP1A1, caffeine; CYP1A2, benzphetamine; CYP3A/2B(?), aniline; 2E1 and TMO; CYP2C9/2E1/3A4); (2) activity generally increases rapidly soon after birth, continues to increase, peaks and then gradually decreases to the adult level (phenytoin; CYP2C9); and (3) activity is high (about the same level as the adult) immediately after birth, decreases and then gradually increases to the adult level (erythromycin; CYP3A4/5). 3. The results of these in vivo and in vitro studies suggest that changes in enzyme activity are due to differences in P450 isoenzymes during development. PMID- 9741959 TI - 3,3'-Diindolylmethane induces CYP1A2 in cultured precision-cut human liver slices. AB - 1. The effect of 3,3'-diindolylmethane (DIM), an indole derivative derived from cruciferous vegetables, on cytochrome P450 (CYP) isoforms in the CYP1A and CYP3A subfamilies has been studied in 72-h cultured human liver slices. 2. In cultured human liver slices 50 microM DIM induced 7-ethoxyresorufin O-deethylase and to a lesser extent 7-methoxyresorufin O-demethylase activities. 3. Western immunoblotting of liver slice microsomes was performed with antibodies to rat CYP1A2 and human CYP3A4. Compared with control liver slice microsomes (dimethyl sulphoxide-only treated), DIM induced levels of CYP1A2 but had little effect on levels of CYP3A4. The treatment of human liver slices with 2 microg/ml of the polycholorinated biphenyl mixture Aroclor 1254 also resulted in an induction of levels of CYP1A2, but had no effect on CYP3A4. 4. These results demonstrate that DIM induces CYP1A isoforms in cultured human liver slices. Some variability in the magnitude of induction of enzyme activities by DIM was observed in four human liver samples examined. For 7-ethoxyresorufin O-deethylase, the magnitude of induction by 50 microM DIM ranged from 2.3- to 19.3-fold. 5. These results demonstrate that cultured human liver slices can be used to evaluate the effect of chemicals derived from cruciferous and other vegetables on CYP isoforms. PMID- 9741960 TI - Lithium: a molecular transducer of mood-stabilization in the treatment of bipolar disorder. PMID- 9741962 TI - Proceedings of the 1st European Union Conference on Glass-Ionomers. 14-16 May 1997. PMID- 9741961 TI - National Institutes of Health Consensus Conference on Breast Cancer Screening for Women Ages 40-49. Proceedings. Bethesda, Maryland, USA. January 21-23, 1997. PMID- 9741963 TI - Spectroscopic studies of 9-hydroxyellipticine binding to DNA. AB - The binding of 9-hydroxyellipticine to calf thymus DNA, poly[d(A-T)]2, and poly[d(G-C)]2 has been studied in detail by means of CD, linear dichroism, resonance light scattering, and molecular dynamics. The transition moment polarizations of 9-hydroxyellipticine were determined in polyvinyl alcohol stretched film. Spectroscopic solution studies of the DNA/drug complex are combined with theoretical CD calculations using the final 50 ps of a series of molecular dynamics simulations as input. The spectroscopic data shows 9 hydroxyellipticine to adopt two main binding modes, one intercalative and the other a stacked binding mode involving the formation of drug oligomers in the DNA major groove. Analysis of the intercalated binding mode in poly[d(A-T)]2 suggests the 9-hydroxyellipticine hydroxyl group lies in the minor groove and hydrogen bonds to water with the pyridine ring protruding into the major groove. The stacked binding mode was examined using resonance light scattering and it was concluded that the drug was forming small oligomer stacks rather than extended aggregates. Reduced linear dichroism measurements suggested a binding geometry that precluded a minor groove binding mode where the plane of the drug makes a 45 degrees angle with the plane of the bases. Thus it was concluded that the drug stacks in the major groove. No obvious differences in the mode of binding of 9 hydroxyellipticine were observed between different DNA sequences; however, the stacked binding mode appeared to be more favorable for calf thymus DNA and poly[d(G-C)]2 than for poly[d(A-T)]2, an observation that could be explained by the slightly greater steric hindrance of the poly[d(A-T)]2 major groove. A strong concentration dependence was observed for the two binding modes where intercalation is favored at very low drug load, with stacking interactions becoming more prominent as the drug concentration is increased. Even at DNA: drug mixing ratios of 70:1 the stacked binding mode was still important for GC-rich DNAs. PMID- 9741964 TI - Proceedings of the 2nd Conference on Foundations of Information Science: The Quest for a Unified Theory of Information. Vienna, Austria, June 11-15, 1996. PMID- 9741965 TI - Protein engineering: Web alert. PMID- 9741966 TI - What should you expect from the study of clinical outcome? PMID- 9741967 TI - Proceedings of The Cape Cod Conference: Sexual Function Assessment in Clinical Trials. Hyannis, Massachusetts, USA. 30-31 May 1997. PMID- 9741968 TI - 1998 winner of Lemelson-MIT $500,000 prize for American innovation. PMID- 9741969 TI - The Genetics of Asthma: Methodological Approaches. UCB Institute of Allergy workshop proceedings. Brussels, Belgium. PMID- 9741970 TI - ISPAD Workshop on Hypoglycemia. Stockholm, Sweden, June 1997. PMID- 9741971 TI - Microbial utilization of the neurotoxin domoic acid: blue mussels (Mytilus edulis) and soft shell clams (Mya arenaria) as sources of the microorganisms. AB - The neurotoxin domoic acid is produced in quantity by the diatom Pseudo-nitzschia multiseries and is released to the environment directly and indirectly via food chains. Presumably there is a mechanism for the biodegradation and disposal of domoic acid and as bacteria are logical candidates for such an activity, a search for bacteria competent to carry out biodegradation of domoic acid was initiated. Extensive trials with a wide variety of bacteria isolated mainly from muds and waters taken from the marine environment showed that the ability to grow on or degrade domoic acid was rare; in fact, domoic acid was inhibitory to resting cells or growing cultures of most of these bacteria. In contrast, using enrichment techniques, it was possible to isolate from molluscan species that eliminate domoic acid readily, i.e., blue mussels (Mytilus edulis) and soft-shell clams (Mya arenaria), bacteria that exhibited growth with and biodegradation of domoic acid when supplemented with low concentrations of growth factors. The species that retain domoic acid for lengthy periods, such as sea scallops (Placopecten magellanicus) and red mussels (Modiolus modiolus), only occasionally yielded bacteria with this capability. The differences may be a result of the mechanisms used by the different shellfish in dealing with domoic acid, i.e., freely available in the blue mussels and soft-shell clams but likely sequestered in the digestive glands of sea scallops and red mussels and thus, largely unavailable for bacterial utilization. The results show that Mytilus edulis and Mya arenaria, almost uniquely, are prime and reliable sources of domoic acid utilizing bacteria. These findings suggest a strong possibility that autochthonous bacteria may be significant factors in the elimination of the neurotoxin in these two species of shellfish. PMID- 9741972 TI - Inactivation of enzymes within spores of Bacillus megaterium ATCC 19213 by hydroperoxides. AB - The organic hydroperoxides t-butyl hydroperoxide, cumene hydroperoxide, and peracetic acid were found to act similarly to hydrogen peroxide in causing inactivation of enzymes within intact spores of bacillus megaterium ATCC 19213 concomitant with mortality. Spores treated with lethal levels of the agents were germinated and permeabilized for enzyme assays. The hierarchy of sensitivities among enolase, glucose-6-phosphate dehydrogenase (G6Pdh), and pyruvate kinase to inactivation varied somewhat with the specific hydroperoxide used, possibly because of the differences in the types of radicals generated. However, each agent inactivated each of the enzymes, albeit at different rates. Comparative assessments of enzyme inactivation by lethal levels of H2O2 or by moist heat showed that some enzymes, such as G6Pdh, are highly sensitive to inactivation, while others, such as ATPases, are much more resistant. The enzymes G6Pdh and aldolase were highly sensitive to hydroperoxide inactivation and also to moist heat, while pyruvate kinase was much more sensitive to hydroperoxides than to moist heat. Our overall interpretation of the findings is that hydroperoxides and moist heat can produce cumulative damage to sensitive enzymes within spores, which progressively diminishes the capacities of the cells to undergo the outgrowth required for return to vegetative life. PMID- 9741973 TI - [Isolation and chemical characterization of type R lipopolysaccharides of a hypovirulent strain of Yersinia pestis]. AB - The lipopolysaccharides LPS I and LPS II, isolated from the hypovirulent EV40 strain of Yersinia pestis, are composed only of type R lipopolysaccharides. This type consists of two forms a and b, depending on their solubility pattern in a solvent mixture containing varying proportions of chloroform, methanol, hexane, and hydrochloric acid. LPS I consists of one subtype, RIb, while LPS II consists of two subtypes, RIIa and RIIb. Analysis by gel electrophoresis shows that the mass of these lipopolysaccharide forms are in the vicinity of 2000-3000 Da. The RIb and RIIb subtypes, which are found in the majority of lipopolysaccharide I and II fractions, are composed of ketoses and amines that are similar to those occurring in LPS I and LPS II. In contrast, the two subtypes RIIa and RIIb are different both in terms of the composition of lipid A and the extent of its substitution. Certain fractions of RIIa contain only lipid A and 3-deoxy-D-manno octulosonic acid (KDO), while other fractions of RIIb possess a lipid A, which is not substituted by arabinose. The whole set of these R-type lipopolysaccharide forms are excellent models for the study of the role of the primary structure of the polysaccharide region, and for the effect of lipid A substitution on the biological activity of bacterial lipopolysaccharides. PMID- 9741975 TI - Association of Clinical Pathologists National Scientific Meeting and 2nd Congress of the European Society of Clinical Pathology. London, United Kingdom, 1-4 October 1997. Abstracts. PMID- 9741974 TI - Recent advances in cyclic nucleotide phosphodiesterases. Part 1 of 2. Proceedings of a symposium. Glasgow, Scotland, July 18-25, 1996. PMID- 9741976 TI - The 10th Scientific Meeting of the European Society for Psychosocial Oncology (ESPO). Stockholm, Sweden, June 14-17, 1998. Abstracts. PMID- 9741977 TI - Vaginoscopical approach to outpatient hysteroscopy. PMID- 9741978 TI - Hepatic abscess. PMID- 9741980 TI - A proactive, data-based determination of the standard of medical care in pediatrics. AB - A 3-week-old infant awoke with a fever. He was taken to the doctor who noted that the child was irritable. The doctor took him to the hospital where a resident performed a spinal tap, started an intravenous (IV) line, and ordered antibiotics. The entire drama, from entering the doctor's office to infusion of ampicillin, took 2 hours. The doctor was sued for malpractice. Expert witnesses for the plaintiff testified that he had deviated from the standard of medical care by taking too long to administer antibiotics, which, in their view, ought to have been given within 30 minutes. Expert witnesses for the defense testified that 2 hours to administer antibiotics in this case was within the standard of care. What ought to be the response of the pediatric community to discrepant expert testimony such as this? One possible response is nothing. Lawyers from both sides will find expert medical witnesses who articulate positions favorable to their clients (as they did in this case), and the truth will emerge after vigorous cross-examination. This, we suggest, is inadequate. We believe that some expert opinions can be viewed as better than others. That is, some opinions describe the standard of medical care correctly while other expert opinions are (to put it charitably) idiosyncratic, failing to depict accurately the skill and care ordinarily administered in comparable situations. Currently, jurors are informed about the standard of care by expert witnesses, who rely on their own medical knowledge and experience. However, a huge body of literature demonstrates that recollections of individual experience are inevitably flawed, and flawed in a nonrandom direction (the Monday morning quarterback phenomenon). Consider the infant with meningitis. When experts in pediatric emergency medicine and pediatric infectious diseases (ID) were asked about the median time from emergency room (ER) presentation to administration of antibiotics in a child with suspected meningitis, their opinions were wrong and slanted toward the outcome known to be desired (namely, a shorter elapsed time). ER physicians (median estimated time to antibiotic administration [AB-TIME] = 46 minutes) and ID physicians (median estimated AB-TIME = 80 minutes) consistently underestimated the actual median value of AB-TIME determined by chart review (120 minutes). From the judicial perspective such potential flaws in expert testimony are assumed to be equally distributed among experts. All admissible evidence is a priori of equal weight until a jury decides otherwise. The standard of medical care is created anew by expert testimony in each individual case, disappearing, like Brigadoon, upon resolution of the dispute. However, to anyone but a lawyer, the standard of medical care must exist as something outside the courtroom testimony of experts, and if it does exist, it should be easily described so that expert testimony can be judged more (or less) accurate in depicting it. We contend that medical care is not a single behavior that conforms to or deviates from an idiosyncratic and retrospectively determined standard, but rather a distribution of behaviors in response to a variety of medical circumstances. For a given scenario, each of several possible responses can be ascribed a relative frequency based on empirical data, and the consequent normal curve depicts the totality of medical care. Substandard care then falls out neatly as behaviors lying outside the large majority of cases. Juries would be empowered (as they are currently) to determine exactly where on this curve substandard care lies, but at least the debate would share the same description of reality. Recent US Supreme Court guidelines regarding expert testimony provide an opportunity to expand the use of databases in medical negligence cases. The Court restricted expert testimony to "scientific knowledge ... based on generating hypotheses and testing them to see if they can be falsified ... (ABSTRACT TRUNCATED) PMID- 9741979 TI - Cyclosporine. PMID- 9741981 TI - A profile of family planning audit in England 1994-1997. AB - Audit has been entered into enthusiastically by most of the multidisciplinary teams giving family planning services but the nature of the subject makes audit particularly difficult in this field. In many cases it has been used as a tool for determining standards rather than a tool for determining compliance to them. If family planning audit and evaluation is to substantially improve in quality, a great deal of work needs to be done, particularly in the formation of appropriate guidelines and in the increased utilisation of audit/research specialists to advise on methodology. It is hoped that the new clinical effectiveness committee of the faculty will take a lead in this; if so it will undoubtedly gain support from faculty members. PMID- 9741982 TI - FAmily planning in Taiwan. PMID- 9741983 TI - Survey of subfertility patients attending a community clinic over a two year span, before and after national folate campaigns. PMID- 9741984 TI - Uptake of family planning services among an ethnically mixed population in a general practice setting. AB - A descriptive study of the uptake of family planning services within an urban general practice which has a mixed ethnic population, was undertaken with the aim of identifying areas where uptake could be improved. The target population was 572 female patients aged between 16 and 45 years during the period August 1995 to July 1996. Of this target population, about 20 per cent had consulted the practice about contraception and four per cent were pregnant. These were excluded. The remaining majority had not consulted the practice about their contraceptive needs. The study looked in more detail at 194 patients who were available for interview, who had not attended the practice for family planning advice during three months from August 1, 1996 to October 31, 1996. Of these, a group could be identified who would have gained benefit from GP involvement. There were 22 'at risk' women who were not using contraception and 11 who had not returned for IUD checks. There was a definite preference for condom use among Asian patients. There was poor uptake of family planning services among 16 to 25 year old Asian patients. Some personal observations and suggestions, for improving uptake and compliance are given. PMID- 9741985 TI - Intimate examinations. PMID- 9741986 TI - Ectopic pregnancy with levonorgestrel releasing intrauterine system. AB - The levonorgestrel releasing intrauterine system has been received with enthusiasm by all involved in women's health for both its contraceptive and non contraceptive uses. This case illustrates a rare event and a possible note of caution. PMID- 9741987 TI - Special issue in memory of Professor Tsujiaki Hata. PMID- 9741989 TI - [Membership list]. PMID- 9741988 TI - Diaphragmatic rupture: suspicion holds the key to prehospital diagnosis. PMID- 9741990 TI - [79th Congress of the Association of Anatomists. Abstracts]. PMID- 9741991 TI - Bone Tumors of the Spine. Proceedings of an international symposium. Bologna, Italy, 3-4 May 1996. PMID- 9741992 TI - 12th International Symposium on Regulatory Peptides. Mackinac Island, Michigan, USA. September 16-20, 1998. Abstracts. PMID- 9741993 TI - Statistical parametric mapping in functional neuroimaging: beyond PET and fMRI activation studies. PMID- 9741994 TI - Tetrofosmin thyroid scintigraphy. PMID- 9741995 TI - Chemotherapy and uptake of technetium-99m sestamibi in breast cancer: reply. PMID- 9741996 TI - Failure to label red blood cells adequately in daily practice using an in vivo method: methodological and clinical considerations [1]. PMID- 9741997 TI - Atypical antipsychotic-like effects of the dopamine D3 receptor agonist, (+)-PD 128,907. AB - Anti-schizophrenia agents with improved efficacy and side-effect profiles are required. A dopamine D3 receptor agonist, R-(+)-trans-3,4a,10b-tetrahydro-4 propyl-2H,5H-[1]benzopyrano[4,3- b]-1,4-oxazin-9-ol HCl ((+)-PD 128,907), displayed an atypical antipsychotic profile comparable to that of clozapine. (+) PD 128,907 blocked stereotypy produced by dizocilpine (MK-801) at 12-fold lower doses than those affecting apomorphine-induced stereotypes in mice and did not produce catalepsy. These effects of (+)-PD 128,907 were stereospecific and were blocked by a D3 antagonist. These data suggest a role for D3 receptors in antipsychotic drug action. PMID- 9741998 TI - Rethinking the approach to cancer: the power of prevention. PMID- 9741999 TI - Image of the month. Pyogenic liver abscess. PMID- 9742000 TI - Triage by flexible sigmoidoscopy: inevitably "short-sighted". PMID- 9742001 TI - Helicobacter pylori and gastric cancer: Koch's postulates fulfilled? PMID- 9742002 TI - The mitochondrial permeability transition: from biochemical curiosity to pathophysiological mechanism. PMID- 9742003 TI - Mutations in the NS5A region of HCV and interferon efficacy. PMID- 9742004 TI - Premature trypsin activation in hereditary pancreatitis. PMID- 9742006 TI - Fasting hyperbilirubinemia. PMID- 9742007 TI - Correcting PT with factor 7 may not improve clotting. PMID- 9742005 TI - Placebo in biliary colic study? PMID- 9742008 TI - Facilitation of intestinal fructose transport. PMID- 9742009 TI - Cysteine protease inhibitor, E-64d, prevents in vitro cerulein- induced trypsinogen activation. PMID- 9742011 TI - Quality Control of Diabetes Care and Chronic Complications in Young People after St. Vincent and Kos. Proceedings of the 4th International Workshop Diabetic Angiopathy in Children. Berlin, Germany, September 4-6, 1997. PMID- 9742010 TI - Allosamidin inhibits the fragmentation of Acremonium chrysogenum but does not influence the cephalosporin-C production of the fungus. AB - The pseudotrisaccharide allosamidin, a potent inhibitor of chitinases, retarded the fragmentation of hyphae but did not affect the fungal growth and cephalosporin-C production in Acremonium chrysogenum. In vitro inhibition of A. chrysogenum cell-bound chitinase(s) by allosamidin revealed that about 47% of the soluble intracellular chitinase activity was resistant to the inhibitory effect of allosamidin. On the other hand, about 76% of the total chitinase activity localised in both the soluble and insoluble enzyme fractions was effectively inhibited by allosamidin. All the chitinase activities were measured using a new procedure based on purified A. chrysogenum chitin as substrate. PMID- 9742012 TI - Human retrovirus implicated in the pathogenesis of IDDM. PMID- 9742013 TI - Proceedings of the 1st European Conference on Immunopharmacology. Berlin, Germany, 26-29 May 1997. PMID- 9742014 TI - Mycobacterium microti: more widespread than previously thought. PMID- 9742015 TI - Serological investigation of a febrile outbreak in Delhi, India, using a rapid immunochromatographic test. PMID- 9742017 TI - Cat scratch disease due to Bartonella henselae serotype Marseille (Swiss cat) in a seronegative patient. PMID- 9742016 TI - Strain from a novel subfamily of hepatitis G virus/hepatitis GB virus C isolated from a Japanese patient: sequence analysis of the envelope 1 region. PMID- 9742018 TI - Jean Alfred Fournier (1832-1914). PMID- 9742019 TI - Issue dedicated to Dr. Robert Traub. PMID- 9742020 TI - 1998 North American Congress of Clinical Toxicology annual meeting. Orlando, Florida, USA. September 9-15, 1998. Abstracts. PMID- 9742021 TI - Medical schools in the United States. PMID- 9742022 TI - Medical schools in Canada. PMID- 9742023 TI - Nephrology's opportunity realized. PMID- 9742024 TI - 1997 Mutation Research Award winner. PMID- 9742025 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 28-1998. A 64-year-old man with cranial-nerve palsies and a positive test for antinuclear cytoplasmic antibodies. PMID- 9742026 TI - Regression of multivalvular regurgitation after the cessation of fenfluramine and phentermine treatment. PMID- 9742027 TI - Fetal DNA and cells in women with systemic sclerosis. PMID- 9742028 TI - Fetal DNA and cells in women with systemic sclerosis. PMID- 9742029 TI - The nephrotic syndrome. PMID- 9742030 TI - The nephrotic syndrome. PMID- 9742031 TI - HIV-protease inhibitors. PMID- 9742032 TI - HIV-protease inhibitors. PMID- 9742033 TI - Role of fecal occult-blood testing. PMID- 9742034 TI - Physician-assisted suicide and euthanasia in the United States. PMID- 9742035 TI - Physician-assisted suicide and euthanasia in the United States. PMID- 9742036 TI - Global distribution of transfusion-transmitted virus. PMID- 9742037 TI - Reforming Medicare: the Gramm plan. PMID- 9742038 TI - Reforming Medicare: the Gramm plan. PMID- 9742039 TI - Telemedicine made easy--the British way. PMID- 9742040 TI - [The Medical Association of Debrecen]. PMID- 9742041 TI - Discoidal niosome based controlled ocular delivery of timolol maleate. AB - Non-ionic surface active agents based discoidal vesicles (discomes) bearing timolol maleate were prepared. Niosomes were incorporated with Solulan C24 in order to effect vesicle to discome transition. The discomes were relatively large in size, 12-60 microm. They were found to entrap a relatively high quantity of timolol maleate. The prepared system characterized for size, shape and drug release profile in vitro. They were found to release the contents following biphasic profile particularly in the case where the drug was loaded using a pH gradient technique. The prepared system could produce or sustain a suitable activity profile upon administration into the ocular cavity; however, systemic absorption was minimized to a negliable level. The discomes were found to be promising and of potential for controlled ocular administration of water-soluble drugs. PMID- 9742043 TI - [Analysis of drinking water: quality control, data collection and report. October 10-11, 1996]. PMID- 9742044 TI - Healing cancer. PMID- 9742045 TI - Proceedings of the NRSP-7/FDA Workshop: Drug Availability for Minor Species in the 21st Century. PMID- 9742042 TI - Characterization of a red beet protein homologous to the essential 36-kilodalton subunit of the yeast V-type ATPase. AB - V-type proton-translocating ATPases (V-ATPases) (EC 3.6.1.3) are electrogenic proton pumps involved in acidification of endomembrane compartments in all eukaryotic cells. V-ATPases from various species consist of 8 to 12 polypeptide subunits arranged into an integral membrane proton pore sector (Vo) and a peripherally associated catalytic sector (V1). Several V-ATPase subunits are functionally and structurally conserved among all species examined. In yeast, a 36-kD peripheral subunit encoded by the yeast (Saccharomyces cerevisiae) VMA6 gene (Vma6p) is required for stable assembly of the Vo sector as well as for V1 attachment. Vma6p has been characterized as a nonintegrally associated Vo subunit. A high degree of sequence similarity among Vma6p homologs from animal and fungal species suggest that this subunit has a conserved role in V-ATPase function. We have characterized a novel Vma6p homolog from red beet (Beta vulgaris) tonoplast membranes. A 44-kD polypeptide cofractionated with V-ATPases upon gel-filtration chromatography of detergent-solubilized tonoplast membranes and was specifically cross-reactive with anti-Vma6p polyclonal antibodies. The 44 kD polypeptide was dissociated from isolated tonoplast preparations by mild chaotropic agents and thus appeared to be nonintegrally associated with the membrane. The putative 44-kD homolog appears to be structurally similar to yeast Vma6p and occupies a similar position within the holoenzyme complex. PMID- 9742046 TI - [Spring conference of the Austrian Cardiological Association. Gmunden, 11-13 June 1998. Abstracts]. PMID- 9742047 TI - Proceedings of the 2nd International Conference on Lacrimal Gland, Tear Film, and Dry Eye Syndromes. Bermuda, November 16-19, 1996. PMID- 9742048 TI - Excerpts from the United States Renal Data System 1998 Annual Data Report. PMID- 9742049 TI - Do kinetics of ADP stimulation of mitochondria really change during myocardial maturation? PMID- 9742050 TI - Why Oedipus and not Christ? PMID- 9742051 TI - Virus taxonomy--San Diego 1998. PMID- 9742052 TI - AHA journals lead with definitive new online site. PMID- 9742053 TI - Familial hypertrophic cardiomyopathy: from mutations to functional defects. AB - Hypertrophic cardiomyopathy is characterized by left and/or right ventricular hypertrophy, which is usually asymmetric and involves the interventricular septum. Typical morphological changes include myocyte hypertrophy and disarray surrounding the areas of increased loose connective tissue. Arrhythmias and premature sudden deaths are common. Hypertrophic cardiomyopathy is familial in the majority of cases and is transmitted as an autosomal-dominant trait. The results of molecular genetics studies have shown that familial hypertrophic cardiomyopathy is a disease of the sarcomere involving mutations in 7 different genes encoding proteins of the myofibrillar apparatus: ss-myosin heavy chain, ventricular myosin essential light chain, ventricular myosin regulatory light chain, cardiac troponin T, cardiac troponin I, alpha-tropomyosin, and cardiac myosin binding protein C. In addition to this locus heterogeneity, there is a wide allelic heterogeneity, since numerous mutations have been found in all these genes. The recent development of animal models and of in vitro analyses have allowed a better understanding of the pathophysiological mechanisms associated with familial hypertrophic cardiomyopathy. One can thus tentatively draw the following cascade of events: The mutation leads to a poison polypeptide that would be incorporated into the sarcomere. This would alter the sarcomeric function that would result (1) in an altered cardiac function and then (2) in the alteration of the sarcomeric and myocyte structure. Some mutations induce functional impairment and support the pathogenesis hypothesis of a "hypocontractile" state followed by compensatory hypertrophy. Other mutations induce cardiac hyperfunction and determine a "hypercontractile" state that would directly induce cardiac hypertrophy. The development of other animal models and of other mechanistic studies linking the genetic mutation to functional defects are now key issues in understanding how alterations in the basic contractile unit of the cardiomyocyte alter the phenotype and the function of the heart. PMID- 9742054 TI - Expression of proto-oncogenes and gene mutation of sarcomeric proteins in patients with hypertrophic cardiomyopathy. AB - Several mutations of cardiac beta-myosin heavy chain (beta-MHC) gene were reported in patients with hypertrophic cardiomyopathy (HCM). Involvement of proto oncogenes has been shown in the mechanism of experimental cardiac hypertrophy. This study sought to examine the effects of c-H-ras and c-myc expression in the steady-state myocardium on hypertrophic changes and to evaluate the possible interaction between beta-MHC mutation and proto-oncogene expression in HCM. Endomyocardial biopsy was performed in 17 HCM patients (5 beta-MHC mutations and 1 troponin T mutation) and 7 control subjects (no mutation). Reverse transcription-polymerase chain reaction analysis revealed c-H-ras expression in all members of both groups. Cardiomyocyte size was correlated with the expression level of c-H-ras (P<0.001), and c-H-ras expression was upregulated in HCM patients (P<0.01). HCM patients with a beta-MHC mutation had the higher c-H-ras expression than did control subjects or patients without a mutation (P<0.01). c myc mRNA was expressed in 7 of 17 HCM patients but not in control subjects. Myocyte size was greater in c-myc-positive HCM patients than in control subjects and c-myc-negative HCM patients (P<0.001 and P<0.05, respectively). The proto oncogene expression did not affect clinical findings, myocardial fibrosis, or disarray. In conclusion, c-H-ras and c-myc expression in the steady-state myocardium may play a role in the hypertrophic mechanism in HCM. It is possible that ss-MHC gene mutation has some effect on the regulation of proto-oncogene expression in HCM. PMID- 9742055 TI - Strong binding of myosin modulates length-dependent Ca2+ activation of rat ventricular myocytes. AB - Reductions in sarcomere length (SL) and concomitant increases in interfilament lattice spacing have been shown to decrease the Ca2+ sensitivity of tension in myocardium. We tested the idea that increased lattice spacing influences the SL dependence of isometric tension by reducing the probability of strong interactions of myosin crossbridges with actin, thereby decreasing cooperative activation of the thin filament. Single ventricular myocytes were isolated by enzymatic digestion of rat hearts and were subsequently rapidly skinned. Maximal tension and Ca2+ sensitivity of tension (ie, pCa50) were measured in the absence and presence of N-ethylmaleimide-modified myosin subfragment 1 (NEM-S1) at both short and long SLs. NEM-S1, a strong-binding non-tension-generating derivative of the myosin head, was applied to single skinned myocytes to cooperatively promote strong binding of endogenous myosin crossbridges. Compared with control myocytes at SL of approximately 1.90 microm, application of NEM-S1 markedly increased submaximal Ca2+-activated tensions and thereby increased Ca2+ sensitivity; ie, pCa50 increased from 5.40+/-0.02 to 5.52+/-0.02 pCa units in the presence of NEM S1. Furthermore, NEM-S1 treatment reversibly eliminated the SL dependence of the Ca2+ sensitivity of tension, in that the DeltapCa50 between short and long lengths was 0. 02+/-0.01 pCa units in the presence of NEM-S1 compared with a DeltapCa50 of 0.10+/-0.01 pCa units in control myocytes. From these results we conclude that the decrease in the Ca2+ sensitivity of tension at short SL results predominantly from decreased cooperative activation of the thin filament due to reductions in the number of strong-binding crossbridges. PMID- 9742056 TI - Thyroid hormone-induced alterations in phospholamban-deficient mouse hearts. AB - Alterations in the expression levels of the sarcoplasmic reticulum (SR) Ca2+ ATPase and its regulator, phospholamban, have been implicated in the effects of thyroxine hormone on cardiac function. To determine the role of phospholamban in these effects, hypothyroidism and hyperthyroidism were induced in phospholamban deficient mice and their isogenic wild types. Hypothyroidism resulted in significant decreases of left ventricular contractility, which could be moderately stimulated by increases in preload or afterload, in both phospholamban deficient and wild-type mice. However, the basal contractile parameters in hypothyroid phospholamban-deficient hearts were at least as high as those exhibited by hyperthyroid wild-type hearts. In hyperthyroidism, there was no further enhancement of the hyperdynamic contractile parameters in phospholamban deficient hearts, although the wild-type hearts exhibited significantly increased contractile function compared with their respective euthyroid groups. Furthermore, increases in preload or afterload did not enhance contractility in either phospholamban-deficient or wild-type hyperthyroid hearts. Examination of the relative tissue levels of cardiac SR Ca2+-ATPase revealed increases in hyperthyroidism and decreases in hypothyroidism compared with euthyroidism, and these changes were similar between phospholamban-deficient and wild-type hearts. An opposite trend was observed for phospholamban expression levels in the wild type group, which were depressed in hyperthyroid hearts but increased in hypothyroid hearts. These findings indicate that (1) thyroid hormones induce similar changes in the cardiac SR Ca2+-ATPase levels in either the presence or absence of phospholamban, (2) the thyroxine-induced increases in SR Ca2+-ATPase levels are not associated with any further stimulation of the hyperdynamic cardiac function in phospholamban-deficient mice, and (3) the decreased contractile parameters in hypothyroid phospholamban-deficient hearts associated with decreases in SR Ca2+-ATPase levels and myosin heavy chain isoform switches are at least as high as those of the stimulated hyperthyroid wild-type hearts. Thus, alterations in the phospholamban level or its activity may be a critical determinant of the contractile responses to altered thyroid states in the mammalian heart. PMID- 9742057 TI - Electrophysiological basis of arrhythmogenicity of QT/T alternans in the long-QT syndrome: tridimensional analysis of the kinetics of cardiac repolarization. AB - Tachycardia-dependent QT/T alternans occurs in patients with the congenital or idiopathic form of long-QT syndrome (LQTS) and may presage the onset of polymorphic ventricular tachyarrhythmias. To examine the electrophysiological basis of arrhythmogenicity of QT/T alternans in LQTS, the tridimensional repolarization pattern of QT/T alternans was studied in the anthopleurin-A model of LQTS, a surrogate for LQT3. In 11 anesthetized mongrel puppies, tridimensional repolarization and activation patterns were analyzed from 256 to 384 unipolar electrograms. Cardiac repolarization was evaluated as the activation-recovery interval (ARI) of local electrograms. To induce QT/T alternans, the pacing cycle length (CL) was abruptly shortened in steps of 50 ms from a basic drive of 1000 ms. ARIs were calculated at epicardial (Epi), midmyocardial (Mid), and endocardial (End) sites. ARI restitution at each site was assessed by using a single premature stimulation delivered after the basic drive. ARI alternans occurred at longer CLs at Mid sites compared with End and Epi sites, and the magnitude of alternans at Mid sites was greater. Two factors contributed to the modulation of ARI during QT/T alternans: (1) differences in restitution kinetics at Mid sites, characterized by larger DeltaARI and a slower time constant (tau), and (2) differences in diastolic intervals resulting in different input to restitution at the same constant CL. These 2 factors could explain not only the onset of alternans at Mid sites at longer CLs but also the critical observation that ARI dispersion between Epi and Mid sites during alternans was greater than during the slower basic CL. Marked ARI alternans could be present in local electrograms without manifest alternation of the QT/T segment in the surface ECG. The latter was seen at critically short CLs associated with reversal of the gradient of ARI between Epi and Mid sites, with a consequent reversal of polarity of the intramyocardial QT wave in alternate cycles. The arrhythmogenicity of QT/T alternans was primarily due to the greater degree of spatial dispersion of repolarization during alternans than during slower rates not associated with alternans. This could result in functional conduction block and reentrant ventricular tachyarrhythmias during the fixed drive associated with alternans. PMID- 9742058 TI - Rapid turnover of connexin43 in the adult rat heart. AB - Remodeling of the distribution of gap junctions is an important feature of anatomic substrates of arrhythmias in patients with healed myocardial infarcts. Mechanisms underlying this process are poorly understood but probably involve changes in gap junction protein (connexin) synthesis, assembly into channels, and degradation. The half-life of the principal cardiac gap junction protein, connexin43 (Cx43), is only 1.5 to 2 hours in primary cultures of neonatal myocytes, but it is unknown whether rapid turnover of Cx43 occurs in the adult heart or is unique to disaggregated neonatal myocytes that are actively reestablishing connections in vitro. To characterize connexin turnover dynamics in the adult heart and to elucidate its potential role in remodeling of gap junctions, we measured Cx43 turnover kinetics and characterized the proteolytic pathways involved in Cx43 degradation in isolated perfused adult rat hearts. Hearts were labeled for 40 minutes with Krebs-Henseleit buffer containing [35S]methionine, and then chase perfusions were performed with nonradioactive buffer for 0, 60, 120, and 240 minutes. Quantitative immunoprecipitation assays of Cx43 radioactivity in 4 hearts at each time point yielded a monoexponential decay curve indicating a Cx43 half-life of 1.3 hours. Proteolytic pathways responsible for Cx43 degradation were elucidated by perfusing isolated rat hearts for 4 hours with specific inhibitors of either lysosomal or proteasomal proteolysis. Immunoblot analysis demonstrated significant increases ( approximately 30%) in Cx43 content in hearts perfused with either lysosomal or proteasomal pathway inhibitors. Most of the Cx43 in hearts perfused with lysosomal inhibitors consisted of phosphorylated isoforms, whereas nonphosphorylated Cx43 accumulated selectively in hearts perfused with a specific proteasomal inhibitor. These results indicate that Cx43 turns over rapidly in the adult heart and is degraded by multiple proteolytic pathways. Regulation of Cx43 degradation could play an important role in gap junction remodeling in response to cardiac injury. PMID- 9742059 TI - Connexin43 is highly localized to sites of disturbed flow in rat aortic endothelium but connexin37 and connexin40 are more uniformly distributed. AB - Vascular endothelial cells are linked by gap junctions, which facilitate the propagation of electrical and chemical signals along the vessel wall. The aim of this study was to determine the distribution and identity of the gap junction structural proteins (connexins) expressed by endothelial cells in situ. Connexin expression in different regions of the rat aortic endothelium was analyzed with the use of indirect immunofluorescence microscopy and Western blotting. Connexin40 and connexin37 were present in most, if not all, of the thoracic and abdominal aortic endothelia in the form of maculae at cell-cell appositions. In contrast, connexin43 was undetectable in most endothelia but extremely abundant in small numbers of cells localized at the downstream edge of the ostia of branching vessels and at flow dividers, regions that experience turbulent shear stress from disturbed blood flow. To examine the relationship of shear stress and connexin43 expression, localized stress was induced by surgical coarctation of the aorta, which was sufficient to cause striking local upregulation of connexin43 within 8 days. Thus, increases in connexin43 levels are an endothelial response to mechanical stress. PMID- 9742060 TI - Hydrogen peroxide decreases pHi in human aortic endothelial cells by inhibiting Na+/H+ exchange. AB - Postischemic endothelial dysfunction may occur as a result of the effects of endogenous oxidants like hydrogen peroxide. Since endothelium-dependent vasodilator function may be affected by pHi, the effect of hydrogen peroxide on endothelial pHi was examined. Hydrogen peroxide (100 micromol/L for 10 minutes) decreased pHi from 7.24+/-0.01 to 7.02+/-0.02 and inhibited recovery from an ammonium chloride-induced intracellular acid load in carboxy SNARF 1 (c-SNARF 1) loaded human aortic endothelial cells in bicarbonate-free solution. Prior inhibition of Na+/H+ exchange with 5-(N-ethyl-N-isopropyl)amiloride (10 micromol/L), by removal of extracellular Na+, or by glycolytic inhibition with iodoacetic acid blocked the subsequent effect of hydrogen peroxide on pHi. A 2 minute exposure to 100 micromol/L H2O2 decreased intracellular ATP levels by approximately 40%; this was prevented by 3-aminobenzamide and nicotinamide (1 mmol/L each), inhibitors of the DNA repair enzyme poly(ADP-ribose) polymerase. Both 3-aminobenzamide and nicotinamide significantly inhibited the hydrogen peroxide-induced intracellular acidification and the effect of hydrogen peroxide on recovery from an intracellular acid load. Hydrogen peroxide decreases pHi in human endothelial cells by inhibiting Na+/H+ exchange. This appears to be mediated by activation of the DNA repair enzyme poly(ADP-ribose) polymerase and subsequent depletion of intracellular ATP. Since a decrease in pHi in this range may alter the activity of NO synthase or affect the synthesis of vasodilator prostaglandins, the effect of hydrogen peroxide on the endothelial Na+/H+ exchanger may be important in the pathogenesis of postischemic endothelial dysfunction. PMID- 9742061 TI - Immune sources of transforming growth factor-beta1 reduce transplant arteriosclerosis: insight derived from a knockout mouse model. AB - Activated CD4-positive T cells are essential in the early stages of arteriosclerotic lesion development after cardiac transplantation. Besides its parenchymal effects, transforming growth factor-beta1 (TGF-beta1) mediates immunosuppressive effects on proliferation and activation of CD4 cells. This study was designed to assess immune contributions of TGF-beta1 to arteriosclerosis by comparing the effect of TGF-beta1-deficient and -competent infiltrating inflammatory cells on the development of intimal thickening in a heterotopic mouse transplant model (CBA to C57B6). Transplant arteriosclerosis was evaluated in cardiac grafts placed into knockout recipients heterozygous for TGF-beta1 (n=7) and was compared with those placed into wild-type recipients (n=11). At 55 days, allografts in TGF-beta1-deficient recipients had increased concentric intimal thickening. Computer-assisted analysis of all elastin-positive vessels (n=173) showed significantly increased luminal occlusion (67.8+/-5.6%) in grafts from TGF-beta1-deficient recipients compared with wild-type recipients (47.4+/-4.1%, P=0.003). To determine whether TGF-beta1 deficiency altered CD4 activation patterns, we studied intragraft cytokine expression. Using 32P-reverse transcriptase polymerase chain reaction assays, we show that TGF-beta1-deficient recipients had an increased expression of the transcription factor STAT 4, interferon gamma, and interleukin-2 (Th1-type response) and unaltered or reduced expression of the transcription factor STAT 6, interleukin-4, and interleukin-10 (Th2-type response). Hence, when present, immune sources of TGF-beta1 attenuate transplant arteriosclerosis. This effect is associated with attenuation of Th1 forces. PMID- 9742062 TI - Caldesmon inhibits active crossbridges in unstimulated vascular smooth muscle: an antisense oligodeoxynucleotide approach. AB - Caldesmon is a thin-filament-associated protein believed to be important in the regulation of smooth muscle contraction, although the precise mechanism is unknown. We used antisense oligodeoxynucleotides to produce intact swine carotid smooth muscle tissue deficient in h-caldesmon. Caldesmon content was decreased by 78% after 7 days in culture with antisense oligodeoxynucleotides but was unchanged in tissues in the presence of sense oligodeoxynucleotides or vehicle. Antisense oligodeoxynucleotides produced a significant decrease in the caldesmon/actin ratio, but no change was measured in the calponin/actin ratio, suggesting that the effect was specific to caldesmon and not other thin-filament associated proteins. Basal and KCl-stimulated levels of myosin light chain phosphorylation were not different among tissues from all 3 groups. In contrast, h-caldesmon-deficient tissues produced 62% less KCl-induced force than controls. Unstimulated h-caldesmon-deficient smooth muscle tissues stretched and then released, redeveloped force, demonstrating active crossbridge cycling; strips containing normal h-caldesmon content did not redevelop force on release. We suggest that in resting vascular smooth muscle, active crossbridges are inhibited by caldesmon. Therefore, regulation of smooth muscle includes a thin-filament based disinhibition component. PMID- 9742064 TI - Gap junction proteins: where they live and how they die. PMID- 9742063 TI - Inhibition of cardiac delayed rectifier K+ current by overexpression of the long QT syndrome HERG G628S mutation in transgenic mice. AB - Mutations in the HERG gene are linked to the LQT2 form of the inherited long-QT syndrome. Transgenic mice were generated expressing high myocardial levels of a particularly severe form of LQT2-associated HERG mutation (G628S). Hearts from G628S mice appeared normal except for a modest enlargement seen only in females. Ventricular myocytes isolated from adult wild-type hearts consistently exhibited an inwardly rectifying E-4031-sensitive K+ current resembling the rapidly activating cardiac delayed rectifier K+ current (Ikr) in its time and voltage dependence; this current was not found in cells isolated from G628S mice. Action potential duration was significantly prolonged in single myocytes from G628S ventricle (cycle length=1 second, 26 degrees C) but not in recordings from intact ventricular strips studied at more physiological rates and temperature (200 to 400 bpm, 37 degrees C). ECG intervals, including QT duration, were unchanged, although minor aberrancies were noted in 20% (16/80) of the G628S mice studied, primarily involving the QRS complex and, more rarely, T-wave morphology. The aberrations were more commonly observed in females than males but could not be correlated with sex-based differences in action potential duration. These results establish the presence of IKr in the adult mouse ventricle and demonstrate the ability of the G628S mutation to exert a dominant negative effect on endogenous IKr in vivo, leading to the expected LQT2 phenotype of prolonged repolarization at the single cell level but not QT prolongation in the intact animal. The model may be useful in dissecting repolarization currents in the mouse heart and as a means of examining the mechanism(s) by which the G628S mutation exerts its dominant negative effect on native cardiac cells in vivo. PMID- 9742065 TI - Correlative microscopy using FluoroNanogold on ultrathin cryosections. Proof of principle. AB - We demonstrate a fluorescent ultrasmall immunogold probe, FluoroNanogold (FNG), to be a versatile reporter system for immunocytochemical labeling of ultrathin cryosections. FNG-labeled molecules in the same ultrathin cryosections can be resolved by two imaging techniques (i.e., fluorescence and electron microscopy). Lactoferrin, a marker protein for the specific granules in human neutrophils, was employed as the target for FNG immunolabeling. The spatial resolution of the fluorescence signal from FNG-labeled specific granules was compatible with that of silver-enhanced gold signal from the same granules in electron microscopy. Our results confirm that FNG can be used as a probe for high-resolution correlation between immunofluorescence and electron microscopy. PMID- 9742066 TI - Nuclear detection of cellular retinoic acid binding proteins I and II with new antibodies. AB - Apart from the retinoic acid nuclear receptor family, there are two low molecular weight (15 kD) cellular retinoic acid binding proteins, named CRABPI and II. Mouse monoclonal and rabbit polyclonal antibodies were raised against these proteins by using as antigens either synthetic peptides corresponding to amino acid sequences unique to CRABPI or CRABPII, or purified CRABP proteins expressed in E. coli. Antibodies specific for mouse and/or human CRABPI and CRABPII were obtained and characterized by immunocytochemistry and immunoblotting. They allowed the detection not only of CRABPI but also of CRABPII in both nuclear and cytosolic extracts from transfected COS-1 cells, mouse embryos, and various cell lines. PMID- 9742067 TI - Spectral morphometric characterization of B-CLL cells versus normal small lymphocytes. AB - Spectral morphometric characterization of typical chronic lymphocytic leukemia (B CLL) cells vs normal small lymphocytes stained by May-Grunwald-Giemsa was carried out by multipixel spectral imaging. The light intensity (450-850 nm of 10(4) pixels) from nuclear domains of each stained cell was recorded and represented as light transmittance spectra and optical density. Transmitted light spectra of two nuclear domains were determined, one with low-intensity light transmittance (LIT) and the other with high-intensity light transmittance (HIT). A spectral library was constructed using the four transmitted light spectra representing the HIT and LIT domains of the normal human lymphocytes and the LIT and HIT domains of the CLL cells. The spectral library served to scan CLL lymphocytes from 10 cases of CLL and the lymphocytes of 10 healthy individuals. Each spectrally similar domain in the nuclei of the lymphocytes was assigned an arbitrary color. The morphometric analysis of the spectrally classified nuclei showed specific spectral patterns for B-CLL in 92% of the cells. The specific spectral characteristics of each of the two cell populations were also observed by their optical density light absorbance spectra. We propose that spectral morphometric analysis may serve as an additional diagnostic tool for detection of CLL lymphocytes in a hematological specimen. PMID- 9742068 TI - Ultrastructural immunolocalization of basic fibroblast growth factor in mast cell secretory granules. Morphological evidence for bfgf release through degranulation. AB - We previously reported that mast cells (MCs) serve as a source of basic fibroblast growth factor (bFGF), a potent angiogenic and mitogenic polypeptide, suggesting that bFGF may mediate MC-related neovascularization and fibroproliferation. Unlike many other growth factors, bFGF lacks a classic peptide sequence for its secretion, and the mechanism(s) for its release remains controversial. Because MCs release a wide spectrum of bioactive products via degranulation, we hypothesized that MC degranulation may be a mechanism of bFGF release and used ultrastructural immunohistochemistry to test the hypothesis. We reasoned that if bFGF is released through degranulation, it should be localized to MC secretory granules. Human tissues with chronic inflammation and rat/mouse tissues with anaphylaxis were studied. In all tissue samples examined, positive staining (or immunogold particle localization) for bFGF in MCs was predominantly in the cytoplasmic granules. Moderate bFGF immunoreactivity was also found in the nucleus, whereas the cytosol and other subcellular organelles exhibited minimal immunogold particle localization. In contrast, no immunogold particle localization for bFGF was observed in lymphocytes or plasma cells. In rat/mouse lingual tissue undergoing anaphylaxis, immunogold particle localization for bFGF was found not only in swollen cytoplasmic granules but also in the extruded granules of MCs. Three different anti-bFGF antibodies gave similar immunogold particle localization patterns, whereas all controls were negative. These results provide morphological evidence suggesting that, despite the lack of a classic secretory peptide in its structure, bFGF is localized to the secretory granules in MCs and may be released through degranulation. PMID- 9742069 TI - A highly sensitive immunofluorescence procedure for analyzing the subcellular distribution of GABAA receptor subunits in the human brain. AB - We designed a protocol to improve the immunohistochemical analysis of human brain structures, which overcomes the limited detection sensitivity, high background, and intense autofluorescence commonly associated with human tissue. This procedure was evaluated by using antibodies against major GABAA receptor subunits (alpha1, alpha2, alpha3, gamma2) in autopsy and surgical specimens. Tissue blocks were briefly fixed by immersion and pretreated with microwave irradiation in sodium citrate buffer. Immunoperoxidase staining revealed a marked enhancement of cell surface immunoreactivity and reduction of background in microwave-irradiated tissue, irrespective of its origin. For confocal laser scanning microscopy, immunofluorescence staining was optimized with the tyramide signal amplification (TSA) technique. This procedure not only dramatically increased the sensitivity for antigen detection but also totally suppressed autofluorescence, thus revealing the cellular and subcellular distribution of GABAA receptor subunits. A distinct neuron-specific expression pattern of the alpha-subunit variants was observed in cerebral cortex and hippocampal formation, along with widespread expression of the gamma2-subunit. Of particular interest was the prominent alpha2 and alpha3-subunit staining on the initial axon segment of pyramidal neurons. This protocol represents a major improvement for high-resolution studies of human brain tissue aimed at investigating morphological alterations underlying neurological diseases. PMID- 9742070 TI - A new quantitative film autoradiographic method of quantifying mRNA transcripts for in situ hybridization. AB - We developed and tested a novel quantitative method for the quantification of film autoradiographs, involving a mathematical model and a dot-blot-based membrane standard scale. The exponential model introduced here, ROD = p1(1 - exp[p2x]), appropriately (r2>0. 999), describes the relation between relative optical density (ROD) and radioactivity (x) in the range between 0 and 240 gray scale values (using a 256-gray scale level digitizer). By means of this model, standard curves with distinct quenching properties can be exactly interconverted, permitting the tissue-equivalent calibration of different standard scales. The membrane standard scale employed here has several advantages, including the flexible radioactivity range, the facile and rapid preparation technique, and the compact size. The feasibility of the quantification procedure is exemplified by the comparative quantification of multiple calmodulin mRNAs in the rat brain by in situ hybridization with [35S]-cRNA probes. The procedure for quantification provides a significant improvement in that the direct and exact comparison of radiolabeled species, even from different experiments, can be reliably performed. Further, the procedure can be adapted to the quantification of autoradiographs produced by other methods. PMID- 9742071 TI - Effect of 3-methylcholanthrene administration on expression of cytochrome P-450 isoforms induced by phenobarbital in rat hepatocytes. AB - The effects of an inducer on expression of cytochrome P-450 (P-450) isoforms induced antecedently by another inducer are unknown. Thus, we examined the amount of phenobarbital (PB)-inducible P-450 isoforms (P-450 2B1/2B2) in hepatocytes from rats injected first with PB and then with 3-methylcholanthrene (MC) (PB+MC treated animals) by quantitative immunohistochemistry. In addition, expression of P-450 2B2 mRNA was examined by in situ hybridization. In PB-treated animals, P 450 2B1/2B2 content increased in perivenular and midzonal hepatocytes. In PB+MC treated animals, however, the PB-induced increase in 2B1/2B2 content was suppressed in perivenular hepatocytes but promoted in midzonal hepatocytes. The hybridization signal for P-450 2B2 mRNA appeared almost exclusively in perivenular hepatocytes after 24 hr of PB injection and disappeared after 48 hr of injection. In PB+MC-treated animals, however, strong hybridization signal was observed in midzonal and perivenular hepatocytes after 48 hr of PB injection. The promotion of the increase in P-450 2B1/2B2 content in midzonal hepatocytes in PB+MC-treated animals probably corresponds to the strong hybridization signal, whereas there appeared to be a divergence between the intensity of the signal and the content in perivenular hepatocytes. The results indicate that MC administration drastically influences the pattern of expression of P-450 isoforms induced by PB in perivenular and midzonal hepatocytes. PMID- 9742072 TI - Immunohistochemical localization of peroxisomal enzymes in developing rat kidney tissues. AB - We studied the developmental changes in the localization of peroxisome-specific enzymes in rat kidney tissues from embryonic Day 16 to postnatal Week 10 by immunoblot analysis and immunohistochemistry, using antibodies for the peroxisomal enzymes catalase, d-amino acid oxidase, l-alpha-hydroxyacid oxidase (isozyme B), and enoyl-CoA hydratase/3-hydroxyacyl-CoA dehydrogenase bifunctional protein. Peroxisomal enzymes were detected in the neonatal kidney by immunoblot analysis and their amount increased with kidney development. By light microscopic immunohistochemistry, they were first localized in a few proximal tubules in the juxtamedullary cortex of 18-day embryos. The distribution of proximal tubules positive for them expanded towards the superficial cortex with development. The full thickness of the cortex became positive for the staining by 14 days after birth. Peroxisomes could be detected by electron microscopy in structurally immature proximal tubules in 18-day embryos. Their size increased and the ultrastructure of subcompartments became clear with continuing development of proximal tubules. These results show that peroxisomal enzymes appear in the immature proximal tubules in the kidney of embryos and that the ultrastructure of the peroxisomes and localization of the peroxisomal enzymes develop along with the maturation of proximal tubules and kidney tissues. PMID- 9742073 TI - Pre-apoptotic alterations in hepatocytes of TNFalpha-treated galactosamine sensitized mice. AB - Tumor necrosis factor (TNF) induces apoptotic death of hepatocytes in the galactosamine (GalN)-sensitized mouse liver after 5 hr. In our study, the most remarkable sign of the early stage of apoptosis was the focal rupture of the outer mitochondrial membrane. Parts of the inner membrane extended through the gap of the outer membrane, whereas the rest of the inner membrane still formed the cristae. This feature appeared in hepatocytes before chromatin condensation. With the diaminobenzidine technique for localization of cytochrome oxidase activity, the reaction product was detectable by light and electron microscopy. Ten percent of the hepatocytes were apoptotic, with condensed chromatin and high enzyme activity, 37% were pre-apoptotic, without chromatin condensation but high enzyme activity, and 53% had neither condensed chromatin nor a remarkable reaction product of cytochrome oxidase activity. Fas (APO-1, CD95) molecules on the plasma membrane of hepatocytes increased and were represented immunohistochemically in cells without chromatin condensation. DNA strand breaks were also detectable before chromatin aggregation. The results of this study indicate that mitochondria play a pivotal role in pre-apoptotic hepatocytes, together with an increase of the Fas molecule on the plasma membrane and with the occurrence of DNA strand breaks in the nucleus. PMID- 9742074 TI - Identification of N- and O-linked oligosaccharides in the human epididymis. AB - The oligosaccharide sequences of glycoconjugates in the human normal epididymis and the nature of linkages were studied with lectin histochemistry. The usual terminal sequences of oligosaccharide side chains in epithelial cell secretions were Neu5Ac2,3Galbeta1,3GalNAc; SO4Galbeta1,3GalNAc; and Galbeta1,4GlcNAc, and they were mainly found in O-linked glycoproteins. The lectin pattern of mitochondria-rich cells differed from that of principal cells. PMID- 9742075 TI - Cortactin localization in actin-containing adult and fetal tissues. AB - Cortactin is a tyrosine kinase substrate that binds to filamentous actin. It represents a highly conserved family of perimembrane signaling proteins. The human homologue of cortactin is encoded by gene EMS1, which is amplified in some human breast, head, and neck carcinomas. This study shows that cortactin is preferentially localized to the apical surfaces of the polarized epithelium, specifically, to the terminal web of small bowel epithelium and to apical surfaces of the proximal renal tubules, thyroid follicles, and bronchiolar epithelium. Cortactin is also seen in cell and tissue types with actin-based contractile capacities, including smooth and striated muscle and myoepithelium. PMID- 9742076 TI - Intracellular retention of the corticotrophin-releasing hormone (CRH) precursor within COS-7 cells. AB - We investigated the intracellular localization of CRH in transiently transfected COS-7 cells expressing the full-length rat corticotropin-releasing hormone (CRH) precursor cDNA. CRH synthesized by transfected COS-7 cells is mainly stored intracellularly. In contrast, CHO-K1 cells expressing the same CRH precursor stored and released equal amounts of immunoreactive (IR)-CRH. Ultrastructural analysis revealed that CRH is stored in electron-dense aggregates in the RER of transiently transfected COS-7 cells and does not migrate into the Golgi apparatus. On the basis of the different intracellular localization, storage, and release of CRH in COS-7 and CHO-K1 cells, we hypothesize that the intracellular trafficking of CRH within the constitutive secretory pathway for protein secretion not only depends on its primary amino acid sequence but might also be influenced by intracellular conditions or factors. PMID- 9742077 TI - A rapid procedure suitable to assess quantitatively the endocytosis of colloidal gold and its conjugates in cultured cells. AB - We measured the endocytic uptake of low-density lipoproteins (LDLs) conjugated to colloidal gold in cultured cells, either by counting gold particles on electron micrographs or by inductively coupled plasma (ICP) mass spectrometry (MS). Both procedures are comparable but the latter requires a considerably shorter time and allows analysis of a much larger sample. In addition, ICP MS, compared to alternative radioactive or fluorescent procedures, offers the major advantage of using the same probe to quantify the endocytic uptake and to follow it by electron microscopy. Therefore, ICP MS analysis provides an easy, rapid, and sensitive quantification of endocytosis that complements the electron microscopic studies. PMID- 9742078 TI - A new immunocytochemical technique for ultrastructural analysis of DNA replication in proliferating cells after application of two halogenated deoxyuridines. AB - We describe a colloidal gold immunolabeling technique for electron microscopy which allows one to differentially visualize portions of DNA replicated during different periods of S-phase. This was performed by incorporating two halogenated deoxyuridines (IdUrd and CldUrd) into Chinese hamster cells and, after cell processing, by detecting them with selected antibodies. This technique, using in particular appropriate blocking solutions and also Tris buffer with a high salt concentration and 1% Tween-20, prevents nonspecific background and crossreaction of both antibodies. Controls such as digestion with DNase and specific staining of DNA with osmium ammine show that labeling corresponds well to replicated DNA. Different patterns of labeling distribution, reflecting different periods of DNA replication during S-phase, were characterized. Cells in early S-phase display a diffuse pattern of labeling with many spots, whereas cells in late S-phase show labeling confined to larger domains, often at the periphery of the nucleus or associated with the nucleolus. The good correlation between our observations and previous double labeling results in immunofluorescence also proved the technique to be reliable. PMID- 9742079 TI - Novel regulatory factors interacting with the promoter of the gene encoding the mRNA cap binding protein (eIF4E) and their function in growth regulation. AB - Regulation of the mRNA cap binding protein (eIF4E) is critical to the control of cellular proliferation since this protein is the rate-limiting factor in translation initiation and transforms fibroblasts and since eIF4E mutants arrest budding yeast in the G1 phase of the cell cycle (cdc33). We previously demonstrated regulation of eIF4E by altered transcription of its mRNA in serum stimulated fibroblasts and in response to c-myc. To identify additional factors regulating eIF4E transcription, we used linker-scanning constructs to characterize sites in the promoter of the eIF4E gene required for its expression. Promoter activity was dependent on sites at -5, -25, -45, and -75; the site at 75 included a previously described myc box. Electrophoretic mobility shift assays identified DNA-protein interactions at -25 and revealed a binding site (TTACCCCCCCTT) that is unique to the eIF4E promoter. Proteins of 68 and 97 kDa bound this site in UV cross-linking and Southwestern experiments. Levels of 4E regulatory factor activities correlated with c-Myc levels, eIF4E expression levels, and protein synthesis in differentiating U937 and HL60 cells, suggesting that these activities may function to regulate protein synthesis rates during differentiation. Since the eIF4E promoter lacked typical TATA and initiator elements, further studies of this novel initiator-homologous element should provide insights into mechanisms of transcription initiation and growth regulation. PMID- 9742080 TI - Point mutations in the WD40 domain of Eed block its interaction with Ezh2. AB - The Polycomb group proteins are involved in maintenance of the silenced state of several developmentally regulated genes. These proteins form large aggregates with different subunit compositions. To explore the nature of these complexes and their function, we used the full-length Eed (embryonic ectoderm development) protein, a mammalian homolog of the Drosophila Polycomb group protein Esc, as a bait in the yeast two-hybrid screen. Several strongly interacting cDNA clones were isolated. The cloned cDNAs all encoded the 150- to 200-amino-acid N-terminal fragment of the mammalian homolog of the Drosophila Enhancer of zeste [E(z)] protein, Ezh2. The full-length Ezh2 bound strongly to Eed in vitro, and Eed coimmunoprecipitated with Ezh2 from murine 70Z/3 cell extracts, confirming the interaction between these proteins observed in yeast. Mutations T1031A and T1040C in one of the WD40 repeats of Eed, which account for the hypomorphic and lethal phenotype of eed in mouse development, blocked binding of Ezh2 to Eed in a two hybrid interaction in yeast and in mammalian cells. These mutations also blocked the interaction between these proteins in vitro. In mammalian cells, the Gal4-Eed fusion protein represses the activity of a promoter bearing Gal4 DNA elements. The N-terminal fragment of the Ezh2 protein abolished the transcriptional repressor activity of Gal4-Eed protein when they were coexpressed in mammalian cells. Eed and Ezh2 were also found to bind RNA in vitro, and RNA altered the interaction between these proteins. These findings suggest that Polycomb group proteins Eed and Ezh2 functionally interact in mammalian cells, an interaction that is mediated by the WD40-containing domain of Eed protein. PMID- 9742081 TI - Biosynthesis and function of the modified DNA base beta-D-glucosyl hydroxymethyluracil in Trypanosoma brucei. AB - beta-D-Glucosyl-hydroxymethyluracil, also called J, is a modified DNA base conserved among kinetoplastid flagellates. In Trypanosoma brucei, the majority of J is present in repetitive DNA but the partial replacement of thymine by J also correlates with transcriptional repression of the variant surface glycoprotein (VSG) genes in the telomeric VSG gene expression sites. To gain a better understanding of the function of J, we studied its biosynthesis in T. brucei and found that it is made in two steps. In the first step, thymine in DNA is converted into hydroxymethyluracil by an enzyme that recognizes specific DNA sequences and/or structures. In the second step, hydroxymethyluracil is glucosylated by an enzyme that shows no obvious sequence specificity. We identified analogs of thymidine that affect the J content of the T. brucei genome upon incorporation into DNA. These analogs were used to study the function of J in the control of VSG gene expression sites. We found that incorporation of bromodeoxyuridine resulted in a 12-fold decrease in J content and caused a partial derepression of silent VSG gene expression site promoters, suggesting that J might strengthen transcriptional repression. Incorporation of hydroxymethyldeoxyuridine, resulting in a 15-fold increase in the J content, caused a reduction in the occurrence of chromosome breakage events sometimes associated with transcriptional switching between VSG gene expression sites in vitro. We speculate that these effects are mediated by the packaging of J containing DNA into a condensed chromatin structure. PMID- 9742082 TI - The nuclear orphan receptor CAR-retinoid X receptor heterodimer activates the phenobarbital-responsive enhancer module of the CYP2B gene. AB - PBREM, the phenobarbital-responsive enhancer module of the cytochrome P-450 Cyp2b10 gene, contains two potential nuclear receptor binding sites, NR1 and NR2. Consistent with the finding that anti-retinoid X receptor (RXR) could supershift the NR1-nuclear protein complex, DNA affinity chromatography with NR1 oligonucleotides enriched the nuclear orphan receptor RXR from the hepatic nuclear extracts of phenobarbital-treated mice. In addition to RXR, the nuclear orphan receptor CAR was present in the same enriched fraction. In the phenobarbital-treated mice, the binding of both CAR and RXR was rapidly increased before the induction of CYP2B10 mRNA. In vitro-translated CAR bound to NR1, but only in the presence of similarly prepared RXR. PBREM was synergistically activated by transfection of CAR and RXR in HepG2 and HEK293 cells when the NR1 site was functional. A CAR-RXR heterodimer has thus been characterized as a trans acting factor for the phenobarbital-inducible Cyp2b10 gene. PMID- 9742083 TI - Mammalian GCN5 and P/CAF acetyltransferases have homologous amino-terminal domains important for recognition of nucleosomal substrates. AB - The yeast transcriptional adapter Gcn5p serves as a histone acetyltransferase, directly linking chromatin modification to transcriptional regulation. Two human homologs of Gcn5p have been reported previously, hsGCN5 and hsP/CAF (p300/CREB binding protein [CBP]-associated factor). While hsGCN5 was predicted to be close to the size of the yeast acetyltransferase, hsP/CAF contained an additional 356 amino-terminal residues of unknown function. Surprisingly, we have found that in mouse, both the GCN5 and the P/CAF genes encode proteins containing this extended amino-terminal domain. Moreover, while a shorter version of GCN5 might be generated upon alternative or incomplete splicing of a longer transcript, mRNAs encoding the longer protein are much more prevalent in both mouse and human cells, and larger proteins are detected by GCN5-specific antisera in both mouse and human cell extracts. Mouse GCN5 (mmGCN5) and mmP/CAF genes are ubiquitously expressed, but maximum expression levels are found in different, complementary sets of tissues. Both mmP/CAF and mmGCN5 interact with CBP/p300. Interestingly, mmGCN5 maps to chromosome 11 and cosegregates with BRCA1, and mmP/CAF maps to a central region of chromosome 17. As expected, recombinant mmGCN5 and mmP/CAF both exhibit histone acetyltransferase activity in vitro with similar substrate specificities. However, in contrast to yeast Gcn5p and the previously reported shorter form of hsGCN5, mmGCN5 readily acetylates nucleosomal substrates as well as free core histones. Thus, the unique amino-terminal domains of mammalian P/CAF and GCN5 may provide additional functions important to recognition of chromatin substrates and the regulation of gene expression. PMID- 9742084 TI - Degradation of myogenic transcription factor MyoD by the ubiquitin pathway in vivo and in vitro: regulation by specific DNA binding. AB - MyoD is a tissue-specific transcriptional activator that acts as a master switch for skeletal muscle differentiation. Its activity is induced during the transition from proliferating, nondifferentiated myoblasts to resting, well differentiated myotubes. Like many other transcriptional regulators, it is a short-lived protein; however, the targeting proteolytic pathway and the underlying regulatory mechanisms involved in the process have remained obscure. It has recently been shown that many short-lived regulatory proteins are degraded by the ubiquitin system. Degradation of a protein by the ubiquitin system proceeds via two distinct and successive steps, conjugation of multiple molecules of ubiquitin to the target protein and degradation of the tagged substrate by the 26S proteasome. Here we show that MyoD is degraded by the ubiquitin system both in vivo and in vitro. In intact cells, the degradation is inhibited by lactacystin, a specific inhibitor of the 26S proteasome. Inhibition is accompanied by accumulation of high-molecular-mass MyoD-ubiquitin conjugates. In a cell-free system, the proteolytic process requires both ATP and ubiquitin and, like the in vivo process, is preceded by formation of ubiquitin conjugates of the transcription factor. Interestingly, the process is inhibited by the specific DNA sequence to which MyoD binds: conjugation and degradation of a MyoD mutant protein which lacks the DNA-binding domain are not inhibited. The inhibitory effect of the DNA requires the formation of a complex between the DNA and the MyoD protein. Id1, which inhibits the binding of MyoD complexes to DNA, abrogates the effect of DNA on stabilization of the protein. PMID- 9742085 TI - Tumor necrosis factor alpha transcription in macrophages is attenuated by an autocrine factor that preferentially induces NF-kappaB p50. AB - Macrophages are a major source of proinflammatory cytokines such as tumor necrosis factor alpha (TNF-alpha), which are expressed during conditions of inflammation, infection, or injury. We identified an activity secreted by a macrophage tumor cell line that negatively regulates bacterial lipopolysaccharide (LPS)-induced expression of TNF-alpha. This activity, termed TNF-alpha-inhibiting factor (TIF), suppressed the induction of TNF-alpha expression in macrophages, whereas induction of three other proinflammatory cytokines (interleukin-1beta [IL 1beta], IL-6, and monocyte chemoattractant protein 1) was accelerated or enhanced. A similar or identical inhibitory activity was secreted by IC-21 macrophages following LPS stimulation. Inhibition of TNF-alpha expression by macrophage conditioned medium was associated with selective induction of the NF kappaB p50 subunit. Hyperinduction of p50 occurred with delayed kinetics in LPS stimulated macrophages but not in fibroblasts. Overexpression of p50 blocked LPS induced transcription from a TNF-alpha promoter reporter construct, showing that this transcription factor is an inhibitor of the TNF-alpha gene. Repression of the TNF-alpha promoter by TIF required a distal region that includes three NF kappaB binding sites with preferential affinity for p50 homodimers. Thus, the selective repression of the TNF-alpha promoter by TIF may be explained by the specific binding of inhibitory p50 homodimers. We propose that TIF serves as a negative autocrine signal to attenuate TNF-alpha expression in activated macrophages. TIF is distinct from the known TNF-alpha-inhibiting factors IL-4, IL 10, and transforming growth factor beta and may represent a novel cytokine. PMID- 9742086 TI - Regulation of Mdm2-directed degradation by the C terminus of p53. AB - The stability of the p53 tumor suppressor protein is regulated by interaction with Mdm2, the product of a p53-inducible gene. Mdm2-targeted degradation of p53 depends on the interaction between the two proteins and is mediated by the proteasome. We show here that in addition to the N-terminal Mdm2 binding domain, the C terminus of p53 participates in the ability of p53 to be degraded by Mdm2. In contrast, alterations in the central DNA binding domain of p53, which change the conformation of the p53 protein, do not abrogate the sensitivity of the protein to Mdm2-mediated degradation. The importance of the C-terminal oligomerization domain to Mdm2-targeted degradation of p53 is likely to reflect the importance of oligomerization of the full-length p53 protein for interaction with Mdm2, as previously shown in vitro. Interestingly, the extreme C-terminal region of p53, outside the oligomerization domain, was also shown to be necessary for efficient degradation, and deletion of this region stabilized the protein without abrogating its ability to bind to Mdm2. Mdm2-resistant p53 mutants were not further stabilized following DNA damage, supporting a role for Mdm2 as the principal regulator of p53 stability in cells. The extreme C terminus of the p53 protein has previously been shown to contain several regulatory elements, raising the possibility that either allosteric regulation of p53 by this domain or interaction between this region and a third protein plays a role in determining the sensitivity of p53 to Mdm2-directed degradation. PMID- 9742088 TI - A novel DNA-binding protein bound to the mitochondrial inner membrane restores the null mutation of mitochondrial histone Abf2p in Saccharomyces cerevisiae. AB - The yeast mitochondrial HMG-box protein, Abf2p, is essential for maintenance of the mitochondrial genome. To better understand the role of Abf2p in the maintenance of the mitochondrial chromosome, we have isolated a multicopy suppressor (YHM2) of the temperature-sensitive defect associated with an abf2 null mutation. The function of Yhm2p was characterized at the molecular level. Yhm2p has 314 amino acid residues, and the deduced amino acid sequence is similar to that of a family of mitochondrial carrier proteins. Yhm2p is localized in the mitochondrial inner membrane and is also associated with mitochondrial DNA in vivo. Yhm2p exhibits general DNA-binding activity in vitro. Thus, Yhm2p appears to be novel in that it is a membrane-bound DNA-binding protein. A sequence that is similar to the HMG DNA-binding domain is important for the DNA-binding activity of Yhm2p, and a mutation in this region abolishes the ability of YHM2 to suppress the temperature-sensitive defect of respiration of the abf2 null mutant. Disruption of YHM2 causes a significant growth defect in the presence of nonfermentable carbon sources such as glycerol and ethanol, and the cells have defects in respiration as determined by 2,3,5,-triphenyltetrazolium chloride staining. Yhm2p may function as a member of the protein machinery for the mitochondrial inner membrane attachment site of mitochondrial DNA during replication and segregation of mitochondrial genomes. PMID- 9742087 TI - Activation of phosphatidylinositol 3-kinase is sufficient for cell cycle entry and promotes cellular changes characteristic of oncogenic transformation. AB - Using a new inducible form of phosphatidylinositol 3-kinase (PI 3-kinase) we have found that PI 3-kinase activation has the following effects on cell growth and proliferation. (i) Activation of PI 3-kinase was sufficient to promote entry into S phase of the cell cycle within several hours. This was shown by activation of cyclin-dependent kinase 4 (Cdk4) and Cdk2 and by the induction of DNA synthesis. (ii) PI 3-kinase activation alone was not, however, sufficient to provide for progression through the entire cell cycle. Instead, prolonged activation of PI 3 kinase in the absence of serum stimulation resulted in apoptosis. It is possible that the cells undergo apoptosis because the PI 3-kinase-induced entry into the cell cycle is abnormal. For example, we found that the cyclin E-Cdk2 complex, which normally disappears after entry into S phase of the cell cycle, fails to be downregulated following induction by PI 3-kinase. (iii) Finally, we found that prolonged activation of PI 3-kinase in the presence of serum resulted in cellular changes that resemble those associated with oncogenic transformation. The cells reached high densities, were irregular and refractile in appearance, and formed colonies in soft agar. In contrast, neither PI 3-kinase nor serum stimulation alone could induce these changes. Our results suggest that activation of PI 3 kinase promotes anchorage-independent cell growth and entry into the cell cycle but does not abrogate the growth factor requirement for cell proliferation. PMID- 9742089 TI - Transcriptional silencing is defined by isoform- and heterodimer-specific interactions between nuclear hormone receptors and corepressors. AB - Nuclear hormone receptors are ligand-regulated transcription factors that play critical roles in metazoan homeostasis, development, and reproduction. Many nuclear hormone receptors exhibit bimodal transcriptional properties and can either repress or activate the expression of a given target gene. Repression appears to require a physical interaction between a receptor and a corepressor complex containing the SMRT/TRAC or N-CoR/RIP13 polypeptides. We wished to better elucidate the rules governing the association of receptors with corepressors. We report here that different receptors interact with different domains in the SMRT and N-CoR corepressors and that these divergent interactions may therefore contribute to distinct repression phenotypes. Intriguingly, different isoforms of a single nuclear hormone receptor class also differ markedly in their interactions with corepressors, indicative of their nonidentical actions in cellular regulation. Finally, we present evidence that combinatorial interactions between different receptors can, through the formation of heterodimeric receptors, result in novel receptor-corepressor interactions not observed for homomeric receptors. PMID- 9742090 TI - Distinct subdomains of human TAFII130 are required for interactions with glutamine-rich transcriptional activators. AB - TFIID is a multiprotein complex consisting of the TATA box binding protein and multiple tightly associated proteins (TAFIIs) that are required for transcription by selected activators. We previously reported cloning and partial characterization of human TAFII130 (hTAFII130). The central domain of hTAFII130 contains four glutamine-rich regions, designated Q1 to Q4, that are involved in interactions with the transcriptional activator Sp1. Mutational analysis has revealed specific regions within the glutamine-rich (Q1 to Q4) central region of hTAFII130 that are required for interaction with distinct activation domains. We tested amino- and carboxyl-terminal deletions of hTAFII130 for interaction with Sp1 activation domains A and B (Sp1A and Sp1B) and the N-terminal activation domain of CREB (CREB-N) by using the yeast two-hybrid system. Our results indicate that Sp1B interacts almost exclusively with the Q1 region of hTAFII130. In contrast, Sp1A makes multiple contacts with Q1 to Q4 of hTAFII130, while CREB N interacts primarily with the Q1-Q2 hTAFII130 subdomain. Consistent with these interaction studies, overexpression of the Q1-to-Q4 region in HeLa cells inhibits Sp1- but not VP16-mediated transcriptional activation. These findings indicate that the Q1-to-Q4 region of hTAFII130 is required for Sp1-mediated transcriptional enhancement in mammalian cells and that different activation domains target distinct subdomains of hTAFII130. PMID- 9742091 TI - Functional promiscuity of gene regulation by serpentine receptors in Dictyostelium discoideum. AB - Serpentine receptors such as smoothened and frizzled play important roles in cell fate determination during animal development. In Dictyostelium discoideum, four serpentine cyclic AMP (cAMP) receptors (cARs) regulate expression of multiple classes of developmental genes. To understand their function, it is essential to know whether each cAR is coupled to a specific gene regulatory pathway or whether specificity results from the different developmental regulation of individual cARs. To distinguish between these possibilities, we measured gene induction in car1 car3 double mutant cell lines that express equal levels of either cAR1, cAR2, or cAR3 under a constitutive promoter. We found that all cARs efficiently mediate both aggregative gene induction by cAMP pulses and induction of postaggregative and prespore genes by persistent cAMP stimulation. Two exceptions to this functional promiscuity were observed. (i) Only cAR1 can mediate adenosine inhibition of cAMP-induced prespore gene expression, a phenomenon that was found earlier in wild-type cells. cAR1's mediation of adenosine inhibition suggests that cAR1 normally mediates prespore gene induction. (ii) Only cAR2 allows entry into the prestalk pathway. Prestalk gene expression is induced by differentiation inducing factor (DIF) but only after cells have been prestimulated with cAMP. We found that DIF-induced prestalk gene expression is 10 times higher in constitutive cAR2 expressors than in constitutive cAR1 or cAR3 expressors (which still have endogenous cAR2), suggesting that cAR2 mediates induction of DIF competence. Since in wild-type slugs cAR2 is expressed only in anterior cells, this could explain the so far puzzling observations that prestalk cells differentiate at the anterior region but that DIF levels are actually higher at the posterior region. After the initial induction of DIF competence, cAMP becomes a repressor of prestalk gene expression. This function can again be mediated by cAR1, cAR2, and cAR3. PMID- 9742092 TI - NDT80 and the meiotic recombination checkpoint regulate expression of middle sporulation-specific genes in Saccharomyces cerevisiae. AB - Distinct classes of sporulation-specific genes are sequentially expressed during the process of spore formation in Saccharomyces cerevisiae. The transition from expression of early meiotic genes to expression of middle sporulation-specific genes occurs at about the time that cells exit from pachytene and form the meiosis I spindle. To identify genes encoding potential regulators of middle sporulation-specific gene expression, we screened for mutants that expressed early meiotic genes but failed to express middle sporulation-specific genes. We identified mutant alleles of RPD3, SIN3, and NDT80 in this screen. Rpd3p, a histone deacetylase, and Sin3p are global modulators of gene expression. Ndt80p promotes entry into the meiotic divisions. We found that entry into the meiotic divisions was not required for activation of middle sporulation genes; these genes were efficiently expressed in a clb1 clb3 clb4 strain, which fails to enter the meiotic divisions due to reduced Clb-dependent activation of Cdc28p kinase. In contrast, middle sporulation genes were not expressed in a dmc1 strain, which fails to enter the meiotic divisions because a defect in meiotic recombination leads to a RAD17-dependent checkpoint arrest. Expression of middle sporulation genes, as well as entry into the meiotic divisions, was restored to a dmc1 strain by mutation of RAD17. Our studies also revealed that NDT80 was a temporally distinct, pre-middle sporulation gene and that its expression was reduced, but not abolished, on mutation of DMC1, RPD3, SIN3, or NDT80 itself. In summary, our data indicate that Ndt80p is required for expression of middle sporulation genes and that the activity of Ndt80p is controlled by the meiotic recombination checkpoint. Thus, middle genes are expressed only on completion of meiotic recombination and subsequent generation of an active form of Ndt80p. PMID- 9742093 TI - Involvement of the tyrosine kinase fer in cell adhesion. AB - The Fer protein belongs to the fes/fps family of nontransmembrane receptor tyrosine kinases. Lack of success in attempts to establish a permanent cell line overexpressing it at significant levels suggested a strong negative selection against too much Fer protein and pointed to a critical cellular function for Fer. Using a tetracycline-regulatable expression system, overexpression of Fer in embryonic fibroblasts was shown to evoke a massive rounding up, and the subsequent detachment of the cells from the substratum, which eventually led to cell death. Induction of Fer expression coincided with increased complex formation between Fer and the cadherin/src-associated substrate p120(cas) and elevated tyrosine phosphorylation of p120(cas). beta-Catenin also exhibited clearly increased phosphotyrosine levels, and Fer and beta-catenin were found to be in complex. Significantly, although the levels of alpha-catenin, beta-catenin, and E-cadherin were unaffected by Fer overexpression, decreased amounts of alpha catenin and beta-catenin were coimmunoprecipitated with E-cadherin, demonstrating a dissolution of adherens junction complexes. A concomitant decrease in levels of phosphotyrosine in the focal adhesion-associated protein p130 was also observed. Together, these results provide a mechanism for explaining the phenotype of cells overexpressing Fer and indicate that the Fer tyrosine kinase has a function in the regulation of cell-cell adhesion. PMID- 9742094 TI - Mutations in RNA polymerase II and elongation factor SII severely reduce mRNA levels in Saccharomyces cerevisiae. AB - Elongation factor SII interacts with RNA polymerase II and enables it to transcribe through arrest sites in vitro. The set of genes dependent upon SII function in vivo and the effects on RNA levels of mutations in different components of the elongation machinery are poorly understood. Using yeast lacking SII and bearing a conditional allele of RPB2, the gene encoding the second largest subunit of RNA polymerase II, we describe a genetic interaction between SII and RPB2. An SII gene disruption or the rpb2-10 mutation, which yields an arrest-prone enzyme in vitro, confers sensitivity to 6-azauracil (6AU), a drug that depresses cellular nucleoside triphosphates. Cells with both mutations had reduced levels of total poly(A)+ RNA and specific mRNAs and displayed a synergistic level of drug hypersensitivity. In cells in which the SII gene was inactivated, rpb2-10 became dominant, as if template-associated mutant RNA polymerase II hindered the ability of wild-type polymerase to transcribe. Interestingly, while 6AU depressed RNA levels in both wild-type and mutant cells, wild-type cells reestablished normal RNA levels, whereas double-mutant cells could not. This work shows the importance of an optimally functioning elongation machinery for in vivo RNA synthesis and identifies an initial set of candidate genes with which SII-dependent transcription can be studied. PMID- 9742096 TI - Requirement of STE50 for osmostress-induced activation of the STE11 mitogen activated protein kinase kinase kinase in the high-osmolarity glycerol response pathway. AB - Exposure of yeast cells to increases in extracellular osmolarity activates the HOG1 mitogen-activated protein (MAP) kinase cascade, which is composed of three tiers of protein kinases: (i) the SSK2, SSK22, and STE11 MAP kinase kinase kinases (MAPKKKs), (ii) the PBS2 MAPKK, and (iii) the HOG1 MAP kinase. Activation of the MAP kinase cascade is mediated by two upstream mechanisms. The SLN1-YPD1 SSK1 two-component osmosensor activates the SSK2 and SSK22 MAPKKKs by direct interaction of the SSK1 response regulator with these MAPKKKs. The second mechanism of HOG1 MAP kinase activation is independent of the two-component osmosensor and involves the SHO1 transmembrane protein and the STE11 MAPKKK. Only PBS2 and HOG1 are common to the two mechanisms. We conducted an exhaustive mutant screening to identify additional elements required for activation of STE11 by osmotic stress. We found that strains with mutations in the STE50 gene, in combination with ssk2Delta ssk22Delta mutations, were unable to induce HOG1 phosphorylation after osmotic stress. Both two-hybrid analyses and coprecipitation assays demonstrated that the N-terminal domain of STE50 binds strongly to the N-terminal domain of STE11. The binding of STE50 to STE11 is constitutive and is not affected by osmotic stress. Furthermore, the two proteins relocalize similarly after osmotic shock. It was concluded that STE50 fulfills an essential role in the activation of the high-osmolarity glycerol response pathway by acting as an integral subunit of the STE11 MAPKKK. PMID- 9742095 TI - Regulation of myogenesis by fibroblast growth factors requires beta-gamma subunits of pertussis toxin-sensitive G proteins. AB - Terminal differentiation of skeletal muscle cells in culture is inhibited by a number of different growth factors whose subsequent intracellular signaling events are poorly understood. In this study, we have investigated the role of heterotrimeric G proteins in mediating fibroblast growth factor (FGF)-dependent signals that regulate myogenic differentiation. Pertussis toxin, which ADP ribosylates and inactivates susceptible G proteins, promotes terminal differentiation in the presence of FGF-2, suggesting that Galpha or Gbeta gamma subunits or both are involved in transducing the FGF-dependent signal(s) that inhibits myogenesis. We found that Gbetagamma subunits are likely to be involved since the expression of the C terminus of beta-adrenergic receptor kinase 1, a Gbetagamma subunit-sequestering agent, promotes differentiation in the presence of FGF-2, and expression of the free Gbeta gamma dimer can replace FGF-2, rescuing cells from pertussis toxin-induced differentiation. Addition of pertussis toxin also blocked FGF-2-mediated activation of mitogen-activated protein kinases (MAPKs). Ectopic expression of dominant active mutants in the Ras/MAPK pathway rescued cells from pertussis toxin-induced terminal differentiation, suggesting that the Gbeta gamma subunits act upstream of the Ras/MAPK pathway. It is unlikely that the pertussis toxin-sensitive pathway is activated by other, as yet unidentified FGF receptors since PDGF (platelet derived growth factor)-stimulated MM14 cells expressing a chimeric receptor containing the FGF receptor-1 intracellular domain and the PDGF receptor extracellular domain were sensitive to pertussis toxin. Our data suggest that FGF mediated signals involved in repression of myogenic differentiation are transduced by a pertussis toxin-sensitive G-protein-coupled mechanism. This signaling pathway requires the action of Gbeta gamma subunits and activation of MAPKs to repress skeletal muscle differentiation. PMID- 9742098 TI - I-PpoI, the endonuclease encoded by the group I intron PpLSU3, is expressed from an RNA polymerase I transcript. AB - PpLSU3, a mobile group I intron in the rRNA genes of Physarum polycephalum, also can home into yeast chromosomal ribosomal DNA (rDNA) (D. E. Muscarella and V. M. Vogt, Mol. Cell. Biol. 13:1023-1033, 1993). By integrating PpLSU3 into the rDNA copies of a yeast strain temperature sensitive for RNA polymerase I, we have shown that the I-PpoI homing endonuclease encoded by PpLSU3 is expressed from an RNA polymerase I transcript. We have also developed a method to integrate mutant forms of PpLSU3 as well as the Tetrahymena intron TtLSU1 into rDNA, by expressing I-PpoI in trans. Analysis of I-PpoI expression levels in these mutants, along with subcellular fractionation of intron RNA, strongly suggests that the full length excised intron RNA, but not RNAs that are further cleaved, serves as or gives rise to the mRNA. PMID- 9742097 TI - Hypersensitivity of Ku-deficient cells toward the DNA topoisomerase II inhibitor ICRF-193 suggests a novel role for Ku antigen during the G2 and M phases of the cell cycle. AB - Ku antigen is a heterodimer, comprised of 86- and 70-kDa subunits, which binds preferentially to free DNA ends. Ku is associated with a catalytic subunit of 450 kDa in the DNA-dependent protein kinase (DNA-PK), which plays a crucial role in DNA double-strand break (DSB) repair and V(D)J recombination of immunoglobulin and T-cell receptor genes. We now demonstrate that Ku86 (86-kDa subunit) deficient Chinese hamster cell lines are hypersensitive to ICRF-193, a DNA topoisomerase II inhibitor that does not produce DSB in DNA. Mutant cells were blocked in G2 at drug doses which had no effect on wild-type cells. Moreover, bypass of this G2 block by caffeine revealed defective chromosome condensation in Ku86-deficient cells. The hypersensitivity of Ku86-deficient cells toward ICRF 193 was not due to impaired in vitro decatenation activity or altered levels of DNA topoisomerase IIalpha or -beta. Rather, wild-type sensitivity was restored by transfection of a Ku86 expression plasmid into mutant cells. In contrast to cells deficient in the Ku86 subunit of DNA-PK, cells deficient in the catalytic subunit of the enzyme neither accumulated in G2/M nor displayed defective chromosome condensation at lower doses of ICRF-193 compared to wild-type cells. Our data suggests a novel role for Ku antigen in the G2 and M phases of the cell cycle, a role that is not related to its role in DNA-PK-dependent DNA repair. PMID- 9742099 TI - Requirements for chromatin modulation and transcription activation by the Pho4 acidic activation domain. AB - Perhaps the best characterized example of an activator-induced chromatin transition is found in the activation of the Saccharomyces cerevisiae acid phosphatase gene PHO5 by the basic helix-loop-helix (bHLH) transcription factor Pho4. Transcription activation of the PHO5 promoter by Pho4 is accompanied by the remodeling of four positioned nucleosomes which is dependent on the Pho4 activation domain but independent of transcription initiation. Whether the requirements for transcription activation through the TATA sequence are different from those necessary for the chromatin transition remains a major outstanding question. In an attempt to understand better the ability of Pho4 to activate transcription and to remodel chromatin, we have initiated a detailed characterization of the Pho4 activation domain. Using both deletion and point mutational analysis, we have defined residues between positions 75 and 99 as being both essential and sufficient to mediate transcription activation. Significantly, there is a marked concordance between the ability of mutations in the Pho4 activation domain to induce chromatin opening and transcription activation. Interestingly, the requirements for transcription activation within the Pho4 activation domain differ significantly if fused to a heterologous bHLH leucine zipper DNA-binding domain. The implications for transcription activation by Pho4 are discussed. PMID- 9742100 TI - Alkylpurine-DNA-N-glycosylase knockout mice show increased susceptibility to induction of mutations by methyl methanesulfonate. AB - Alkylpurine-DNA-N-glycosylase (APNG) null mice have been generated by homologous recombination in embryonic stem cells. The null status of the animals was confirmed at the mRNA level by reverse transcription-PCR and by the inability of cell extracts of tissues from the knockout (ko) animals to release 3 methyladenine (3-meA) or 7-methylguanine (7-meG) from 3H-methylated calf thymus DNA in vitro. Following treatment with DNA-methylating agents, increased persistence of 7-meG was found in liver sections of APNG ko mice in comparison with wild-type (wt) mice, demonstrating an in vivo phenotype for the APNG null animals. Unlike other null mutants of the base excision repair pathway, the APNG ko mice exhibit a very mild phenotype, show no outward abnormalities, are fertile, and have an apparently normal life span. Neither a difference in the number of leukocytes in peripheral blood nor a difference in the number of bone marrow polychromatic erythrocytes was found when ko and wt mice were exposed to methylating or chloroethylating agents. These agents also showed similar growth inhibitory effects in primary embryonic fibroblasts isolated from ko and wt mice. However, treatment with methyl methanesulfonate resulted in three- to fourfold more hprt mutations in splenic T lymphocytes from APNG ko mice than in those from wt mice. These mutations were predominantly single-base-pair changes; in the ko mice, they consisted primarily of AT-->TA and GC-->TA transversions, which most likely are caused by 3-meA and 3- or 7-meG, respectively. These results clearly show an important role for APNG in attenuating the mutagenic effects of N alkylpurines in vivo. PMID- 9742102 TI - A sequence of the CIS gene promoter interacts preferentially with two associated STAT5A dimers: a distinct biochemical difference between STAT5A and STAT5B. AB - Two distinct genes encode the closely related signal transducer and activator of transcription proteins STAT5A and STAT5B. The molecular mechanisms of gene regulation by STAT5 and, particularly, the requirement for both STAT5 isoforms are still undetermined. Only a few STAT5 target genes, among them the CIS (cytokine-inducible SH2-containing protein) gene, have been identified. We cloned the human CIS gene and studied the human CIS gene promoter. This promoter contains four STAT binding elements organized in two pairs. By electrophoretic mobility shift assay studies using nuclear extracts of UT7 cells stimulated with erythropoietin, we showed that these four sequences bound to STAT5-containing complexes that exhibited different patterns and affinities: the three upstream STAT binding sequences bound to two distinct STAT5-containing complexes (C0 and C1) and the downstream STAT box bound only to the slower-migrating C1 band. Using nuclear extracts from COS-7 cells transfected with expression vectors for the prolactin receptor, STAT5A, and/or STAT5B, we showed that the C1 complex was composed of a STAT5 tetramer and was dependent on the presence of STAT5A. STAT5B lacked this property and bound with a stronger affinity than did STAT5A to the four STAT sequences as a homodimer (C0 complex). This distinct biochemical difference between STAT5A and STAT5B was confirmed with purified activated STAT5 recombinant proteins. Moreover, we showed that the presence on the same side of the DNA helix of a second STAT sequence increased STAT5 binding and that only half of the palindromic STAT binding sequence was sufficient for the formation of a STAT5 tetramer. Again, STAT5A was essential for this cooperative tetrameric association. This property distinguishes STAT5A from STAT5B and could be essential to explain the transcriptional regulation diversity of STAT5. PMID- 9742101 TI - Identification of a novel cortactin SH3 domain-binding protein and its localization to growth cones of cultured neurons. AB - Cortactin is an actin-binding protein that contains several potential signaling motifs including a Src homology 3 (SH3) domain at the distal C terminus. Translocation of cortactin to specific cortical actin structures and hyperphosphorylation of cortactin on tyrosine have been associated with the cortical cytoskeleton reorganization induced by a variety of cellular stimuli. The function of cortactin in these processes is largely unknown in part due to the lack of information about cellular binding partners for cortactin. Here we report the identification of a novel cortactin-binding protein of approximately 180 kDa by yeast two-hybrid interaction screening. The interaction of cortactin with this 180-kDa protein was confirmed by both in vitro and in vivo methods, and the SH3 domain of cortactin was found to direct this interaction. Since this protein represents the first reported natural ligand for the cortactin SH3 domain, we designated it CortBP1 for cortactin-binding protein 1. CortBP1 contains two recognizable sequence motifs within its C-terminal region, including a consensus sequence for cortactin SH3 domain-binding peptides and a sterile alpha motif. Northern and Western blot analysis indicated that CortBP1 is expressed predominately in brain tissue. Immunofluorescence studies revealed colocalization of CortBP1 with cortactin and cortical actin filaments in lamellipodia and membrane ruffles in fibroblasts expressing CortBP1. Colocalization of endogenous CortBP1 and cortactin was also observed in growth cones of developing hippocampal neurons, implicating CortBP1 and cortactin in cytoskeleton reorganization during neurite outgrowth. PMID- 9742103 TI - ADR1-mediated transcriptional activation requires the presence of an intact TFIID complex. AB - The yeast transcriptional activator ADR1, which is required for ADH2 and other genes' expression, contains four transactivation domains (TADs). While previous studies have shown that these TADs act through GCN5 and ADA2, and presumably TFIIB, other factors are likely to be involved in ADR1 function. In this study, we addressed the question of whether TFIID is also required for ADR1 action. In vitro binding studies indicated that TADI of ADR1 was able to retain TAFII90 from yeast extracts and TADII could retain TBP and TAFII130/145. TADIV, however, was capable of retaining multiple TAFIIs, suggesting that TADIV was binding TFIID from yeast whole-cell extracts. The ability of TADIV truncation derivatives to interact with TFIID correlated with their transcription activation potential in vivo. In addition, the ability of LexA-ADR1-TADIV to activate transcription in vivo was compromised by a mutation in TAFII130/145. ADR1 was found to associate in vivo with TFIID in that immunoprecipitation of either TAFII90 or TBP from yeast whole-cell extracts specifically coimmunoprecipitated ADR1. Most importantly, depletion of TAFII90 from yeast cells dramatically reduced ADH2 derepression. These results indicate that ADR1 physically associates with TFIID and that its ability to activate transcription requires an intact TFIID complex. PMID- 9742104 TI - The deafness-associated mitochondrial DNA mutation at position 7445, which affects tRNASer(UCN) precursor processing, has long-range effects on NADH dehydrogenase subunit ND6 gene expression. AB - The pathogenetic mechanism of the deafness-associated mitochondrial DNA (mtDNA) T7445C mutation has been investigated in several lymphoblastoid cell lines from members of a New Zealand pedigree exhibiting the mutation in homoplasmic form and from control individuals. We show here that the mutation flanks the 3' end of the tRNASer(UCN) gene sequence and affects the rate but not the sites of processing of the tRNA precursor. This causes an average reduction of approximately 70% in the tRNASer(UCN) level and a decrease of approximately 45% in protein synthesis rate in the cell lines analyzed. The data show a sharp threshold in the capacity of tRNASer(UCN) to support the wild-type protein synthesis rate, which corresponds to approximately 40% of the control level of this tRNA. Strikingly, a 7445 mutation-associated marked reduction has been observed in the level of the mRNA for the NADH dehydrogenase (complex I) ND6 subunit gene, which is located approximately 7 kbp upstream and is cotranscribed with the tRNASer(UCN) gene, with strong evidence pointing to a mechanistic link with the tRNA precursor processing defect. Such reduction significantly affects the rate of synthesis of the ND6 subunit and plays a determinant role in the deafness-associated respiratory phenotype of the mutant cell lines. In particular, it accounts for their specific, very significant decrease in glutamate- or malate-dependent O2 consumption. Furthermore, several homoplasmic mtDNA mutations affecting subunits of NADH dehydrogenase may play a synergistic role in the establishment of the respiratory phenotype of the mutant cells. PMID- 9742105 TI - Coactivator TIF1beta interacts with transcription factor C/EBPbeta and glucocorticoid receptor to induce alpha1-acid glycoprotein gene expression. AB - The transcription of the alpha1-acid glycoprotein gene is induced by inflammatory cytokines and glucocorticoids. C/EBPbeta is a major transcription factor involved in the induction of the agp gene by some cytokines. In this report, we have identified a novel transcriptional intermediary factor, TIF1beta, which could enhance the transcription of the agp gene by the glucocorticoid receptor (GR) and C/EBPbeta. TIF1beta belongs to a subgroup of RING (really interesting new gene) finger proteins that contain a RING finger preceding two B box-type fingers and a putative coiled-coil domain (RBCC domain). Immunoprecipitation experiments showed that the interaction between GR and TIF1beta is ligand independent. The overexpression of the TIF1beta gene enhances GR-regulated expression in a ligand- and glucocorticoid-responsive element (GRE)-dependent manner. TIF1beta can also augment C/EBPbeta-mediated activity on wild-type and GRE-mutated agp genes, but this augmentation is diminished when all three C/EBPbeta-binding elements are mutated. Functional and biochemical characterizations indicated that the bZIP domain of C/EBPbeta and the RBCC domain, plant homeodomain finger, and bromodomain of TIF1beta are crucial for the interactions of these proteins. Taken together, these results suggest that TIF1beta serves as a converging mediator of signal transduction pathways of glucocorticoids and some inflammatory cytokines. PMID- 9742107 TI - Molecular determinants of NF-kappaB-inducing kinase action. AB - NF-kappaB corresponds to an inducible eukaryotic transcription factor complex that is negatively regulated in resting cells by its physical assembly with a family of cytoplasmic ankyrin-rich inhibitors termed IkappaB. Stimulation of cells with various proinflammatory cytokines, including tumor necrosis factor alpha (TNF-alpha), induces nuclear NF-kappaB expression. TNF-alpha signaling involves the recruitment of at least three proteins (TRADD, RIP, and TRAF2) to the type 1 TNF-alpha receptor tail, leading to the sequential activation of the downstream NF-kappaB-inducing kinase (NIK) and IkappaB-specific kinases (IKKalpha and IKKbeta). When activated, IKKalpha and IKKbeta directly phosphorylate the two N-terminal regulatory serines within IkappaB alpha, triggering ubiquitination and rapid degradation of this inhibitor in the 26S proteasome. This process liberates the NF-kappaB complex, allowing it to translocate to the nucleus. In studies of NIK, we found that Thr-559 located within the activation loop of its kinase domain regulates NIK action. Alanine substitution of Thr-559 but not other serine or threonine residues within the activation loop abolishes its activity and its ability to phosphorylate and activate IKKalpha. Such a NIK-T559A mutant also dominantly interferes with TNF-alpha induction of NF-kappaB. We also found that ectopically expressed NIK both spontaneously forms oligomers and displays a high level of constitutive activity. Analysis of a series of NIK deletion mutants indicates that multiple subregions of the kinase participate in the formation of these NIK-NIK oligomers. NIK also physically assembles with downstream IKKalpha; however, this interaction is mediated through a discrete C-terminal domain within NIK located between amino acids 735 and 947. When expressed alone, this C terminal NIK fragment functions as a potent inhibitor of TNF-alpha-mediated induction of NF-kappaB and alone is sufficient to disrupt the physical association of NIK and IKKalpha. Together, these findings provide new insights into the molecular basis for TNF-alpha signaling, suggesting an important role for heterotypic and possibly homotypic interactions of NIK in this response. PMID- 9742106 TI - Protein kinase C-delta is an important signaling molecule in insulin-like growth factor I receptor-mediated cell transformation. AB - To investigate the potential role of protein kinase C-delta (PKC-delta) in insulin-like growth factor I receptor (IGF-IR)-mediated cell transformation, an oncogenic gag-IGF-IR beta-fusion receptor lacking the entire extracellular domain, which was designated NM1, and a full-length IGF-IR were coexpressed with either wild-type PKC-delta (PKC-deltaWT) or an ATP-binding mutant of PKC-delta (PKC-deltaK376R) in NIH 3T3 fibroblasts. While overexpression of PKC-deltaWT did not affect NM1- and IGF-IR-induced focus and colony formation of NIH 3T3 cells, expression of PKC-deltaK376R severely impaired these events. In contrast, NM1 mediated cell growth in monolayer was not affected by coexpressing PKC deltaK376R. PKC-deltaWT and PKC-deltaK376R were constitutively phosphorylated on a tyrosine residue(s) in the NM1- and IGF-IR-expressing cells and were associated with them in an IGF-I-independent manner. Activated IGF-IR was able to phosphorylate purified PKC-delta in vitro and stimulated its kinase activity. Furthermore, the level of endogenous PKC-delta protein was up-regulated through transcriptional activation in response to long-term IGF-IR activation. Taken together, our results demonstrate that PKC-delta plays an important role in IGF IR-mediated cell transformation, probably via association of the receptor with PKC-delta and its activation through protein up-regulation and tyrosine phosphorylation. Competition with endogenous PKC-delta for NM1 and IGF-IR association by PKC-deltaK376R is probably an important mechanism underlying the PKC-deltaK376R-mediated inhibition of cell transformation by NM1 and IGF-IR. PMID- 9742108 TI - Productive and nonproductive complexes of Ku and DNA-dependent protein kinase at DNA termini. AB - DNA-dependent protein kinase (DNA-PK) is the only eukaryotic protein kinase known to be specifically activated by double-stranded DNA (dsDNA) termini, accounting for its importance in repair of dsDNA breaks and its role in physiologic processes involving dsDNA breaks, such as V(D)J recombination. In this study we conducted kinase and binding analyses using DNA-PK on DNA termini of various lengths in the presence and absence of Ku. We confirmed our previous observations that DNA-PK can bind DNA termini in the absence of Ku, and we determined rate constants for binding. However, in the presence of Ku, DNA-PK can assume either a productive or a nonproductive configuration, depending on the length of the DNA terminus. For dsDNA greater than 26 bp, the productive mode is achieved and Ku increases the affinity of the DNA-PK for the Ku:DNA complex. The change in affinity is achieved by increases in both the kinetic association rate and reduction in the kinetic dissociation rate. For dsDNA smaller than 26 bp, the nonproductive mode, in which DNA-PK is bound to Ku:DNA but is inactive as a kinase, is assumed. Both the productive and nonproductive configurations are likely to be of physiologic importance, depending on the distance of the dsDNA break site to other protein complexes, such as nucleosomes. PMID- 9742109 TI - Inhibition of PrKX, a novel protein kinase, and the cyclic AMP-dependent protein kinase PKA by the regulatory proteins of adeno-associated virus type 2. AB - Adeno-associated virus encodes four nonstructural proteins, which are known as Rep78, Rep68, Rep52, and Rep40. Expression of these nonstructural proteins affects cell growth and gene expression through processes that have not yet been characterized. Using a yeast two-hybrid screen, we have demonstrated that a stable interaction occurs between the viral proteins Rep78 and Rep52 and the putative protein kinase PrKX, which is encoded on the X chromosome. The stability and specificity of the Rep-PrKX interaction were confirmed by coimmunoprecipitation of complexes assembled in vitro and in vivo. Overexpressed PrKX, which was purified from cos cells, was shown to phosphorylate a synthetic protein kinase A (PKA) substrate. However, this activity was dramatically inhibited by stoichiometric amounts of Rep52 and weakly inhibited with Rep68, which lacks the carboxy-terminal sequence contained in Rep52. Similarly, a stable interaction was observed with Rep78, which also contains the carboxy-terminal sequence of Rep52. A stable interaction and inhibition were also observed between Rep52 and the catalytic subunit of PKA. By using surface plasmon resonance and kinetic studies, Kis of approximately 300 and 167 nM were calculated for Rep52 with PKA and with PrKX, respectively. Thus, Rep52 but not Rep68 can significantly inhibit the trans- and autophosphorylation activities of these kinases. The biological effects of Rep78-specific inhibition of PKA-responsive genes are illustrated by the reduction of steady-state levels of cyclic AMP-responsive element-binding protein and cyclin A protein. PMID- 9742110 TI - Influence of intron length on alternative splicing of CD44. AB - Although the splicing of transcripts from most eukaryotic genes occurs in a constitutive fashion, some genes can undergo a process of alternative splicing. This is a genetically economical process which allows a single gene to give rise to several protein isoforms by the inclusion or exclusion of sequences into or from the mature mRNA. CD44 provides a unique example; more than 1,000 possible isoforms can be produced by the inclusion or exclusion of a central tandem array of 10 alternatively spliced exons. Certain alternatively spliced exons have been ascribed specific functions; however, independent regulation of the inclusion or skipping of each of these exons would clearly demand an extremely complex regulatory network. Such a network would involve the interaction of many exon specific trans-acting factors with the pre-mRNA. Therefore, to assess whether the exons are indeed independently regulated, we have examined the alternative exon content of a large number of individual CD44 cDNA isoforms. This analysis shows that the downstream alternatively spliced exons are favored over those lying upstream and that alternative exons are often included in blocks rather than singly. Using a novel in vivo alternative splicing assay, we show that intron length has a major influence upon the alternative splicing of CD44. We propose a kinetic model in which short introns may overcome the poor recognition of alternatively spliced exons. These observations suggest that for CD44, intron length has been exploited in the evolution of the genomic structure to enable tissue-specific patterns of splicing to be maintained. PMID- 9742111 TI - Processivity of the Saccharomyces cerevisiae poly(A) polymerase requires interactions at the carboxyl-terminal RNA binding domain. AB - The interaction of the Fip1 subunit of polyadenylation factor I with the Saccharomyces cerevisiae poly(A) polymerase (PAP) was assayed in vivo by two hybrid analysis and was found to involve two separate regions on PAP, located at opposite ends of the protein sequence. In vitro, Fip1 blocks access of the RNA primer to an RNA binding site (RBS) that overlaps the Fip1 carboxy-terminal interaction region and, in doing so, shifts PAP to a distributive mode of action. Partial truncation of this RBS has the same effect, indicating that this site is required for processivity. A comparison of the utilization of ribo- and deoxyribonucleotides as substrates indicates the existence on PAP of a second RBS which recognizes the last three nucleotides at the 3' end of the primer. This site discriminates against deoxyribonucleotides at the 3' end, and interactions at this site are not affected by Fip1. Further analysis revealed that the specificity of PAP for adenosine is not simply a function of the ATP binding site but also reflects interactions with bases at the 3' end of the primer and at another contact site 14 nucleotides upstream of the 3' end. These results suggest that the unique specificity of PAP for ribose and base, and thus the extent and type of activity with different substrates, depends on interactions at multiple nucleotide binding sites. PMID- 9742112 TI - Functional activity of the fanconi anemia protein FAA requires FAC binding and nuclear localization. AB - Fanconi anemia (FA) is an autosomal recessive disease characterized by genomic instability, cancer susceptibility, and cellular hypersensitivity to DNA-cross linking agents. Eight complementation groups of FA (FA-A through FA-H) have been identified. Two FA genes, corresponding to complementation groups FA-A and FA-C, have been cloned, but the functions of the encoded FAA and FAC proteins remain unknown. We have recently demonstrated that FAA and FAC interact to form a nuclear complex. In this study, we have analyzed a series of mutant forms of the FAA protein with respect to functional activity, FAC binding, and nuclear localization. Mutation or deletion of the amino-terminal nuclear localization signal (NLS) of FAA results in loss of functional activity, loss of FAC binding, and cytoplasmic retention of FAA. Replacement of the NLS sequence with a heterologous NLS sequence, derived from the simian virus 40 T antigen, results in nuclear localization but does not rescue functional activity or FAC binding. Nuclear localization of the FAA protein is therefore necessary but not sufficient for FAA function. Mutant forms of FAA which fail to bind to FAC also fail to promote the nuclear accumulation of FAC. In addition, wild-type FAC promotes the accumulation of wild-type FAA in the nucleus. Our results suggest that FAA and FAC perform a concerted function in the cell nucleus, required for the maintenance of chromosomal stability. PMID- 9742113 TI - myc boxes, which are conserved in myc family proteins, are signals for protein degradation via the proteasome. AB - Cellular levels of the rapidly degraded c-myc protein play an important role in determining the proliferation status of cells. Increased levels of c-myc are frequently associated with rapidly proliferating tumor cells. We show here that myc boxes I and II, found in the N termini of all members of the myc protein family, function to direct the degradation of the c-myc protein. Both myc boxes I and II contain sufficient information to independently direct the degradation of otherwise stably expressed proteins to which they are fused. At least part of the myc box-directed degradation occurs via the proteasome. The mechanism of myc box directed degradation appears to be conserved between yeast and mammalian cells. Our results suggest that the myc boxes may play an important role in regulating the level and activity of the c-myc protein. PMID- 9742114 TI - Transcriptional regulation of the SMK1 mitogen-activated protein kinase gene during meiotic development in Saccharomyces cerevisiae. AB - Meiotic development (sporulation) in Saccharomyces cerevisiae is characterized by an ordered pattern of gene expression, with sporulation-specific genes classified as early, middle, mid-late, or late depending on when they are expressed. SMK1 encodes a mitogen-activated protein kinase required for spore morphogenesis that is expressed as a middle sporulation-specific gene. Here, we identify the cis acting DNA elements that regulate SMK1 transcription and characterize the phenotypes of mutants with altered expression patterns. The SMK1 promoter contains an upstream activating sequence (UASS) that specifically interacts with the transcriptional activator Abf1p. The Abf1p-binding sites from the early HOP1 and the middle SMK1 promoters are functionally interchangeable, demonstrating that these elements do not play a direct role in their differential transcriptional timing. Timing of SMK1 expression is determined by another cis acting DNA sequence termed MSE (for middle sporulation element). The MSE is required not only for activation of SMK1 transcription during middle sporulation but also for its repression during vegetative growth and early meiosis. In addition, the SMK1 MSE can repress vegetative expression in the context of the HOP1 promoter and convert HOP1 from an early to a middle gene. SMK1 function is not contingent on its tight transcriptional regulation as a middle sporulation specific gene. However, promoter mutants with different quantitative defects in SMK1 transcript levels during middle sporulation show distinct sporulation phenotypes. PMID- 9742115 TI - Dominant-negative mutations in the G-protein-coupled alpha-factor receptor map to the extracellular ends of the transmembrane segments. AB - G-protein-coupled receptors (GPCRs) transduce the signals for a wide range of hormonal and sensory stimuli by activating a heterotrimeric guanine nucleotide binding protein (G protein). The analysis of loss-of-function and constitutively active receptor mutants has helped to reveal the functional properties of GPCRs and their role in human diseases. Here we describe the identification of a new class of mutants, dominant-negative mutants, for the yeast G-protein-coupled alpha-factor receptor (Ste2p). Sixteen dominant-negative receptor mutants were isolated based on their ability to inhibit the response to mating pheromone in cells that also express wild-type receptors. Detailed analysis of two of the strongest mutant receptors showed that, unlike other GPCR interfering mutants, they were properly localized at the plasma membrane and did not alter the stability or localization of wild-type receptors. Furthermore, their dominant negative effect was inversely proportional to the relative amount of wild-type receptors and was reversed by overexpressing the G-protein subunits, suggesting that these mutants compete with the wild-type receptors for the G protein. Interestingly, the dominant-negative mutations are all located at the extracellular ends of the transmembrane segments, defining a novel region of the receptor that is important for receptor signaling. Altogether, our results identify residues of the alpha-factor receptor specifically involved in ligand binding and receptor activation and define a new mechanism by which GPCRs can be inactivated that has important implications for the evaluation of receptor mutations in other G-protein-coupled receptors. PMID- 9742116 TI - The locus control region is necessary for gene expression in the human beta globin locus but not the maintenance of an open chromatin structure in erythroid cells. AB - Studies in many systems have led to the model that the human beta-globin locus control region (LCR) regulates the transcription, chromatin structure, and replication properties of the beta-globin locus. However the precise mechanisms of this regulation are unknown. We have developed strategies to use homologous recombination in a tissue culture system to examine how the LCR regulates the locus in its natural chromosomal environment. Our results show that when the functional components of the LCR, as defined by transfection and transgenic studies, are deleted from the endogenous beta-globin locus in an erythroid background, transcription of all beta-globin genes is abolished in every cell. However, formation of the remaining hypersensitive site(s) of the LCR and the presence of a DNase I-sensitive structure of the beta-globin locus are not affected by the deletion. In contrast, deletion of 5'HS5 of the LCR, which has been suggested to serve as an insulator, has only a minor effect on beta-globin transcription and does not influence the chromatin structure of the locus. These results show that the LCR as currently defined is not necessary to keep the locus in an "open" conformation in erythroid cells and that even in an erythroid environment an open locus is not sufficient to permit transcription of the beta like globin genes. PMID- 9742118 TI - Association of guide RNA binding protein gBP21 with active RNA editing complexes in Trypanosoma brucei. AB - RNA editing in Trypanosoma brucei mitochondria produces mature mRNAs by a series of enzyme-catalyzed reactions that specifically insert or delete uridylates in association with a macromolecular complex. Using a mitochondrial fraction enriched for in vitro RNA editing activity, we produced several monoclonal antibodies that are specific for a 21-kDa guide RNA (gRNA) binding protein initially identified by UV cross-linking. Immunofluorescence studies localize the protein to the mitochondrion, with a preference for the kinetoplast. The antibodies cause a supershift of previously identified gRNA-specific ribonucleoprotein complexes and immunoprecipitate in vitro RNA editing activities that insert and delete uridylates. The immunoprecipitated material also contains gRNA-specific endoribonuclease, terminal uridylyltransferase, and RNA ligase activities as well as gRNA and both edited and unedited mRNA. The immunoprecipitate contains numerous proteins, of which the 21-kDa protein, a 90 kDa protein, and novel 55- and 16-kDa proteins can be UV cross-linked to gRNA. These studies indicate that the 21-kDa protein associates with the ribonucleoprotein complex (or complexes) that catalyze RNA editing. PMID- 9742117 TI - Mechanistic principles in NR box-dependent interaction between nuclear hormone receptors and the coactivator TIF2. AB - Nuclear hormone receptors exert transcriptional activation of target genes upon hormone induction via interactions with the basal transcription machinery. This interaction is mediated by cofactors which physically bind to receptors, thereby acting as coactivators or corepressors leading to activation or repression, respectively. Here we report the screening for and cloning of a peroxisome proliferator receptor-interacting protein, the rat homolog of TIF2. By sequence comparison with the related coactivator SRC-1, we identified three short conserved motifs (NR boxes) in both proteins which are the putative binding sites of TIF2 to nuclear hormone receptors. We demonstrate here by generation of amino acid exchanges within the NR boxes that all three boxes located in the receptor interaction domain of TIF2 are necessary and sufficient for interaction. The three boxes individually can bind to hormone receptors but display preferences in binding for certain receptors. In addition, we show that the interaction domain of TIF2 can compete with other AF-2-dependent cofactors for binding to receptors. Finally, we demonstrate cooperative binding of two TIF2 molecules to a heterodimeric nuclear receptor complex even in the presence of only one cognate ligand, indicating an allosteric effect on the heterodimeric partner upon coactivator binding. PMID- 9742119 TI - A positive GATA element and a negative vitamin D receptor-like element control atrial chamber-specific expression of a slow myosin heavy-chain gene during cardiac morphogenesis. AB - We have used the slow myosin heavy chain (MyHC) 3 gene to study the molecular mechanisms that control atrial chamber-specific gene expression. Initially, slow MyHC 3 is uniformly expressed throughout the tubular heart of the quail embryo. As cardiac development proceeds, an anterior-posterior gradient of slow MyHC 3 expression develops, culminating in atrial chamber-restricted expression of this gene following chamberization. Two cis elements within the slow MyHC 3 gene promoter, a GATA-binding motif and a vitamin D receptor (VDR)-like binding motif, control chamber-specific expression. The GATA element of the slow MyHC 3 is sufficient for expression of a heterologous reporter gene in both atrial and ventricular cardiomyocytes, and expression of GATA-4, but not Nkx2-5 or myocyte enhancer factor 2C, activates reporter gene expression in fibroblasts. Equivalent levels of GATA-binding activity were found in extracts of atrial and ventricular cardiomyocytes from embryonic chamberized hearts. These observations suggest that GATA factors positively regulate slow MyHC 3 gene expression throughout the tubular heart and subsequently in the atria. In contrast, an inhibitory activity, operating through the VDR-like element, increased in ventricular cardiomyocytes during the transition of the heart from a tubular to a chambered structure. Overexpression of the VDR, acting via the VDR-like element, duplicates the inhibitory activity in ventricular but not in atrial cardiomyocytes. These data suggest that atrial chamber-specific expression of the slow MyHC 3 gene is achieved through the VDR-like inhibitory element in ventricular cardiomyocytes at the time distinct atrial and ventricular chambers form. PMID- 9742121 TI - Haploinsufficiency of MSX1: a mechanism for selective tooth agenesis. AB - Previously, we found that the cause of autosomal dominant selective tooth agenesis in one family is a missense mutation resulting in an arginine-to-proline substitution in the homeodomain of MSX1. To determine whether the tooth agenesis phenotype may result from haploinsufficiency or a dominant-negative mechanism, we have performed biochemical and functional analyses of the mutant protein Msx1(R31P). We show that Msx1(R31P) has perturbed structure and reduced thermostability compared with wild-type Msx1. As a consequence, the biochemical activities of Msx1(R31P) are severely impaired, since it exhibits little or no ability to interact with DNA or other protein factors or to function in transcriptional repression. We also show that Msx1(R31P) is inactive in vivo, since it does not display the activities of wild-type Msx1 in assays of ectopic expression in the limb. Furthermore, Msx1(R31P) does not antagonize the activity of wild-type Msx1 in any of these assays. Because Msx1(R31P) appears to be inactive and does not affect the action of wild-type Msx1, we propose that the phenotype of affected individuals with selective tooth agenesis is due to haploinsufficiency. PMID- 9742120 TI - The AD1 and AD2 transactivation domains of E2A are essential for the antiapoptotic activity of the chimeric oncoprotein E2A-HLF. AB - The chimeric oncoprotein E2A-HLF, generated by the t(17;19) chromosomal translocation in pro-B-cell acute lymphoblastic leukemia, incorporates the transactivation domains of E2A and the basic leucine zipper (bZIP) DNA-binding and protein dimerization domain of HLF (hepatic leukemic factor). The ability of E2A-HLF to prolong the survival of interleukin-3 (IL-3)-dependent murine pro-B cells after IL-3 withdrawal suggests that it disrupts signaling pathways normally responsible for cell suicide, allowing the cells to accumulate as transformed lymphoblasts. To determine the structural motifs that contribute to this antiapoptotic effect, we constructed a panel of E2A-HLF mutants and programmed their expression in IL-3-dependent murine pro-B cells (FL5.12 line), using a zinc inducible vector. Neither the E12 nor the E47 product of the E2A gene nor the wild-type HLF protein was able to protect the cells from apoptosis induced by IL 3 deprivation. Surprisingly, different combinations of disabling mutations within the HLF bZIP domain had little effect on the antiapoptotic property of the chimeric protein, so long as the amino-terminal portion of E2A remained intact. In the context of a bZIP domain defective in DNA binding, mutants retaining either of the two transactivation domains of E2A were able to extend cell survival after growth factor deprivation. Thus, the block of apoptosis imposed by E2A-HLF in pro-B lymphocytes depends critically on the transactivating regions of E2A. Since neither DNA binding nor protein dimerization through the bZIP domain of HLF is required for this effect, we propose mechanisms whereby protein-protein interactions with the amino-terminal region of E2A allow the chimera to act as a transcriptional cofactor to alter the expression of genes regulating the apoptotic machinery in pro-B cells. PMID- 9742122 TI - E1B 19,000-molecular-weight protein interacts with and inhibits CED-4-dependent, FLICE-mediated apoptosis. AB - Genetic studies of the nematode Caenorhabditis elegans (C. elegans) have identified several important components of the cell death pathway, most notably CED-3, CED-4, and CED-9. CED-4 directly interacts with the Bcl-2 homologue CED-9 (or the mammalian Bcl-2 family member Bcl-xL) and the caspase CED-3 (or the mammalian caspases ICE and FLICE). This trimolecular complex of CED-4, CED-3, and CED-9 is functional in that CED-9 inhibits CED-4 from activating CED-3 and thereby inhibits apoptosis in heterologous systems. The E1B 19,000-molecular weight protein (E1B 19K) is a potent apoptosis inhibitor and the adenovirus homologue of Bcl-2-related apoptosis inhibitors. Since E1B 19K and Bcl-xL have functional similarity, we determined if E1B 19K interacts with CED-4 and regulates CED-4-dependent caspase activation. Binding analysis indicated that E1B 19K interacts with CED-4 in a Saccharomyces cerevisiae two-hybrid assay, in vitro, and in mammalian cell lysates. The subcellular localization pattern of CED 4 was dramatically changed by E1B 19K, supporting the theory of a functional interaction between CED-4 and E1B 19K. Whereas expression of CED-4 alone could not induce cell death, coexpression of CED-4 and FLICE augmented cell death induction by FLICE, which was blocked by expression of E1B 19K. Even though E1B 19K did not prevent FLICE-induced apoptosis, it did inhibit CED-4-dependent, FLICE-mediated apoptosis, which suggested that CED-4 was required for E1B 19K to block FLICE activation. Thus, E1B 19K functions through interacting with CED-4, and presumably a mammalian homologue of CED-4, to inhibit caspase activation and apoptosis. PMID- 9742123 TI - FGF-18, a novel member of the fibroblast growth factor family, stimulates hepatic and intestinal proliferation. AB - The fibroblast growth factors (FGFs) play key roles in controlling tissue growth, morphogenesis, and repair in animals. We have cloned a novel member of the FGF family, designated FGF-18, that is expressed primarily in the lungs and kidneys and at lower levels in the heart, testes, spleen, skeletal muscle, and brain. Sequence comparison indicates that FGF-18 is highly conserved between humans and mice and is most homologous to FGF-8 among the FGF family members. FGF-18 has a typical signal sequence and was glycosylated and secreted when it was transfected into 293-EBNA cells. Recombinant murine FGF-18 protein (rMuFGF-18) stimulated proliferation in the fibroblast cell line NIH 3T3 in vitro in a heparan sulfate dependent manner. To examine its biological activity in vivo, rMuFGF-18 was injected into normal mice and ectopically overexpressed in transgenic mice by using a liver-specific promoter. Injection of rMuFGF-18 induced proliferation in a wide variety of tissues, including tissues of both epithelial and mesenchymal origin. The two tissues which appeared to be the primary targets of FGF-18 were the liver and small intestine, both of which exhibited histologic evidence of proliferation and showed significant gains in organ weight following 7 (sometimes 3) days of FGF-18 treatment. Transgenic mice that overexpressed FGF-18 in the liver also exhibited an increase in liver weight and hepatocellular proliferation. These results suggest that FGF-18 is a pleiotropic growth factor that stimulates proliferation in a number of tissues, most notably the liver and small intestine. PMID- 9742124 TI - Biased suppression of hematopoiesis and multiple developmental defects in chimeric mice containing Shp-2 mutant cells. AB - Shp-2 is a cytoplasmic tyrosine phosphatase that contains two Src homology 2 (SH2) domains at the N terminus. Biochemical data suggests that Shp-2 acts downstream of a variety of receptor and cytoplasmic tyrosine kinases. A targeted deletion mutation in the N-terminal SH2 (SH2-N) domain results in embryonic lethality of homozygous mutant mice at midgestation. In vitro embryonic stem (ES) cell differentiation assays suggest that Shp-2 might play an important role in hematopoiesis. By aggregating homozygous mutant (Shp-2(-/-)) ES cells and wild type (WT) embryos, we created Shp-2(-/-)-WT chimeric animals. We report here an essential role of Shp-2 in the control of blood cell development. Despite the widespread contribution of mutant cells to various tissues, no Shp-2(-/-) progenitors for erythroid or myeloid cells were detected in the fetal liver and bone marrow of chimeric animals by using the in vitro CFU assay. Furthermore, hematopoiesis was defective in Shp-2(-/-) yolk sacs. In addition, the Shp-2 mutation caused multiple developmental defects in chimeric mice, characterized by short hind legs, aberrant limb features, split lumbar vertebrae, abnormal rib patterning, and pathological changes in the lungs, intestines, and skin. These results demonstrate a functional involvement of Shp-2 in the differentiation of multiple tissue-specific cells and in body organization. More importantly, the requirement for Shp-2 is more stringent in hematopoiesis than in other systems. PMID- 9742125 TI - Mutagenesis of the BH3 domain of BAX identifies residues critical for dimerization and killing. AB - The BCL-2 family of proteins is comprised of proapoptotic as well as antiapoptotic members (S. N. Farrow and R. Brown, Curr. Opin. Genet. Dev. 6:45 49, 1996). A prominent death agonist, BAX, forms homodimers and heterodimerizes with multiple antiapoptotic members. Death agonists have an amphipathic alpha helix, called BH3; however, the initial assessment of BH3 in BAX has yielded conflicting results. Our BAX deletion constructs and minimal domain constructs indicated that the BH3 domain was required for BAX homodimerization and heterodimerization with BCL-2, BCL-XL, and MCL-1. An extensive site-directed mutagenesis of BH3 revealed that substitutions along the hydrophobic face of BH3, especially charged substitutions, had the greatest affects on dimerization patterns and death agonist activity. Particularly instructive was the BAX mutant mIII-1 (L63A, G67A, L70A, and M74A), which replaced the hydrophobic face of BH3 with alanines, preserving its amphipathic nature. BAXmIII-1 failed to form heterodimers or homodimers by yeast two-hybrid or immunoprecipitation analysis yet retained proapoptotic activity. This suggests that BAX's killing function reflects mechanisms beyond its binding to BCL-2 or BCL-XL to inhibit them or simply displace other protein partners. Notably, BAXmIII-1 was found predominantly in mitochondrial membranes, where it was homodimerized as assessed by homobifunctional cross-linkers. This characteristic of BAXmIII-1 correlates with its capacity to induce mitochondrial dysfunction, caspase activation, and apoptosis. These data are consistent with a model in which BAX death agonist activity may require an intramembranous conformation of this molecule that is not assessed accurately by classic binding assays. PMID- 9742127 TI - Sld2, which interacts with Dpb11 in Saccharomyces cerevisiae, is required for chromosomal DNA replication. AB - The DPB11 gene, which genetically interacts with DNA polymerase II (epsilon), one of three replicative DNA polymerases, is required for DNA replication and the S phase checkpoint in Saccharomyces cerevisiae. To identify factors interacting with Dbp11, we have isolated sld (synthetically lethal with dpb11-1) mutations which fall into six complementation groups (sld1 to -6). In this study, we characterized SLD2, encoding an essential 52-kDa protein. High-copy SLD2 suppressed the thermosensitive growth defect caused by dpb11-1. Conversely, high copy DPB11 suppressed the temperature-sensitive growth defect caused by sld2-6. The interaction between Sld2 and Dpb11 was detected in a two-hybrid assay. This interaction was evident at 25 degreesC but not at 34 degreesC when Sld2-6 or Dpb11-1 replaced its wild-type protein. No interaction between Sld2-6 and Dpb11-1 could be detected even at 25 degreesC. Immunoprecipitation experiments confirmed that Dpb11 physically interacts with Sld2. sld2-6 cells were defective in DNA replication at the restrictive temperature, as were dpb11-1 cells. Further, in dpb11-1 and sld2-6 cells, the bubble-shaped replication intermediates formed in the region of the autonomously replicating sequence reduced quickly after a temperature shift. These results strongly suggest the involvement of the Dpb11 Sld2 complex in a step close to the initiation of DNA replication. PMID- 9742126 TI - ErbB tyrosine kinases and the two neuregulin families constitute a ligand receptor network. AB - The recently isolated second family of neuregulins, NRG2, shares its primary receptors, ErbB-3 and ErbB-4, and induction of mammary cell differentiation with NRG1 isoforms, suggesting functional redundancy of the two growth factor families. To address this possibility, we analyzed receptor specificity of NRGs by using an engineered cellular system. The activity of isoform-specific but partly overlapping patterns of specificities that collectively activate all eight ligand-stimulatable ErbB dimers was revealed. Specifically, NRG2-alpha [corrected], like NRG1-beta [corrected], emerges as a narrow-specificity ligand, whereas NRG2-beta [corrected] is a pan-ErbB ligand that binds with different affinities to all receptor combinations, including those containing ErbB-1, but excluding homodimers of ErbB-2. The latter protein, however, displayed cooperativity with the direct NRG receptors. Apparently, signaling by all NRGs is funneled through the mitogen-activated protein kinase (MAPK). However, the duration and potency of MAPK activation depend on the identity of the stimulatory ligand-receptor ternary complex. We conclude that the NRG-ErbB network represents a complex and nonredundant machinery developed for fine-tuning of signal transduction. PMID- 9742128 TI - Functional characterization of the N terminus of Sir3p. AB - Silent information regulator 3 is an essential component of the Saccharomyces cerevisiae silencing complex that functions at telomeres and the silent mating type loci, HMR and HML. We show that expression of the N- and C-terminal-encoding halves of SIR3 in trans partially complements the mating defect of the sir3 null allele, suggesting that the two domains have distinct functions. We present here a functional characterization of these domains. The N-terminal domain (Sir3N) increases both the frequency and extent of telomere-proximal silencing when expressed ectopically in SIR+ yeast strains, although we are unable to detect interaction between this domain and any known components of the silencing machinery. In contrast to its effect at telomeres, Sir3N overexpression derepresses transcription of reporter genes inserted in the ribosomal DNA (rDNA) array. Immunolocalization of Sir3N-GFP and Sir2p suggests that Sir3N directly antagonizes nucleolar Sir2p, releasing an rDNA-bound population of Sir2p so that it can enhance repression at telomeres. Overexpression of the C-terminal domain of either Sir3p or Sir4p has a dominant-negative effect on telomeric silencing. In strains overexpressing the C-terminal domain of Sir4p, elevated expression of either full-length Sir3p or Sir3N restores repression and the punctate pattern of Sir3p and Rap1p immunostaining. The similarity of Sir3N and Sir3p overexpression phenotypes suggests that Sir3N acts as an allosteric effector of Sir3p, either enhancing its interactions with other silencing components or liberating the full length protein from nonfunctional complexes. PMID- 9742129 TI - Telomere length regulation and telomeric chromatin require the nonsense-mediated mRNA decay pathway. AB - Rap1p localization factor 4 (RLF4) is a Saccharomyces cerevisiae gene that was identified in a screen for mutants that affect telomere function and alter the localization of the telomere binding protein Rap1p. In rlf4 mutants, telomeric silencing is reduced and telomere DNA tracts are shorter, indicating that RLF4 is required for both the establishment and/or maintenance of telomeric chromatin and for the control of telomere length. In this paper, we demonstrate that RLF4 is allelic to NMD2/UPF2, a gene required for the nonsense-mediated mRNA decay (NMD) pathway (Y. Cui, K. W. Hagan, S. Zhang, and S. W. Peltz, Mol. Cell. Biol. 9:423 436, 1995, and F. He and A. Jacobson, Genes Dev. 9:437-454, 1995). The NMD pathway, which requires Nmd2p/Rlf4p together with two other proteins, (Upf1p and Upf3p), targets nonsense messages for degradation in the cytoplasm by the exoribonuclease Xrn1p. Deletion of UPF1 and UPF3 caused telomere-associated defects like those caused by rlf4 mutations, implying that the NMD pathway, rather than an NMD-independent function of Nmd2p/Rlf4p, is required for telomere functions. In addition, telomere length regulation required Xrn1p but not Rat1p, a nuclear exoribonuclease with functional similarity to Xrn1p (A. W. Johnson, Mol. Cell. Biol. 17:6122-6130, 1997). In contrast, telomere-associated defects were not observed in pan2, pan3, or pan2 pan3 strains, which are defective in the intrinsic deadenylation-dependent decay of normal (as opposed to nonsense) mRNAs. Thus, loss of the NMD pathway specifically causes defects at telomeres, demonstrating a physiological requirement for the NMD pathway in normal cell functions. We propose a model in which the NMD pathway regulates the levels of specific mRNAs that are important for telomere functions. PMID- 9742130 TI - Identification of rCop-1, a new member of the CCN protein family, as a negative regulator for cell transformation. AB - By using a model system for cell transformation mediated by the cooperation of the activated H-ras oncogene and the inactivated p53 tumor suppressor gene, rCop 1 was identified by mRNA differential display as a gene whose expression became lost after cell transformation. Homology analysis indicates that rCop-1 belongs to an emerging cysteine-rich growth regulator family called CCN, which includes connective-tissue growth factor, CYR61, CEF10 (v-src inducible), and the product of the nov proto-oncogene. Unlike the other members of the CCN gene family, rCop 1 is not an immediate-early gene, it lacks the conserved C-terminal domain which was shown to confer both growth-stimulating and heparin-binding activities, and its expression is lost in cells transformed by a variety of mechanisms. Ectopic expression of rCop-1 by retroviral gene transfers led to cell death in a transformation-specific manner. These results suggest that rCop-1 represents a new class of CCN family proteins that have functions opposing those of the previously identified members. PMID- 9742131 TI - Circadian regulation of a Drosophila homolog of the mammalian Clock gene: PER and TIM function as positive regulators. AB - The Clock gene plays an essential role in the manifestation of circadian rhythms (approximately 24 h) in mice and is a member of the basic helix-loop-helix (bHLH) PER-ARNT-SIM (PAS) superfamily of transcription factors. Here we report the characterization of a novel Drosophila bHLH-PAS protein that is highly homologous to mammalian CLOCK. (Similar findings were recently described by Allada et al. Cell 93:791-804, 1998, and Darlington et al., Science 280:1599-1603, 1998.) Transcripts from this putative Clock ortholog (designated dClock) undergo daily rhythms in abundance that are antiphase to the cycling observed for the RNA products from the Drosophila melanogaster circadian clock genes period (per) and timeless (tim). Furthermore, dClock RNA cycling is abolished and the levels are at trough values in the absence of either PER or TIM, suggesting that these two proteins can function as transcriptional activators, a possibility which is in stark contrast to their previously characterized role in transcriptional autoinhibition. Finally, the temporal regulation of dClock expression is quickly perturbed by shifts in light-dark cycles, indicating that this molecular rhythm is closely connected to the photic entrainment pathway. The isolation of a Drosophila homolog of Clock together with the recent discovery of mammalian homologs of per indicate that there is high structural conservation in the integral components underlying circadian oscillators in Drosophila and mammals. Nevertheless, because mammalian Clock mRNA is constitutively expressed, our findings are a further example of striking differences in the regulation of putative circadian clock orthologs in different species. PMID- 9742132 TI - Modifications of the 5' cap of mRNAs during Xenopus oocyte maturation: independence from changes in poly(A) length and impact on translation. AB - The translation of specific maternal mRNAs is regulated during early development. For some mRNAs, an increase in translational activity is correlated with cytoplasmic extension of their poly(A) tails; for others, translational inactivation is correlated with removal of their poly(A) tails. Recent results in several systems suggest that events at the 3' end of the mRNA can affect the state of the 5' cap structure, m7G(5')ppp(5')G. We focus here on the potential role of cap modifications on translation during early development and on the question of whether any such modifications are dependent on cytoplasmic poly(A) addition or removal. To do so, we injected synthetic RNAs into Xenopus oocytes and examined their cap structures and translational activities during meiotic maturation. We draw four main conclusions. First, the activity of a cytoplasmic guanine-7-methyltransferase increases during oocyte maturation and stimulates translation of an injected mRNA bearing a nonmethylated GpppG cap. The importance of the cap for translation in oocytes is corroborated by the sensitivity of protein synthesis to cap analogs and by the inefficient translation of mRNAs bearing nonphysiologically capped 5' termini. Second, deadenylation during oocyte maturation does not cause decapping, in contrast to deadenylation-triggered decapping in Saccharomyces cerevisiae. Third, the poly(A) tail and the N-7 methyl group of the cap stimulate translation synergistically during oocyte maturation. Fourth, cap ribose methylation of certain mRNAs is very inefficient and is not required for their translational recruitment by poly(A). These results demonstrate that polyadenylation can cause translational recruitment independent of ribose methylation. We propose that polyadenylation enhances translation through at least two mechanisms that are distinguished by their dependence on ribose modification. PMID- 9742133 TI - Dendritic hyperpolarization-activated currents modify the integrative properties of hippocampal CA1 pyramidal neurons. AB - Step hyperpolarizations evoked slowly activating, noninactivating, and slowly deactivating inward currents from membrane patches recorded in the cell-attached patch configuration from the soma and apical dendrites of hippocampal CA1 pyramidal neurons. The density of these hyperpolarization-activated currents (Ih) increased over sixfold from soma to distal dendrites. Activation curves demonstrate that a significant fraction of Ih channels is active near rest and that the range is hyperpolarized relatively more in the distal dendrites. Ih activation and deactivation kinetics are voltage-and temperature-dependent, with time constants of activation and deactivation decreasing with hyperpolarization and depolarization, respectively. Ih demonstrated a mixed Na+-K+ conductance and was sensitive to low concentrations of external CsCl. Dual whole-cell recordings revealed regional differences in input resistance (Rin) and membrane polarization rates (taumem) across the somatodendritic axis that are attributable to the spatial gradient of Ih channels. As a result of these membrane effects the propagation of subthreshold voltage transients is directionally specific. The elevated dendritic Ih density decreases EPSP amplitude and duration and reduces the time window over which temporal summation takes place. The backpropagation of action potentials into the dendritic arborization was impacted only slightly by dendritic Ih, with the most consistent effect being a decrease in dendritic action potential duration and an increase in afterhyperpolarization. Overall, Ih acts to dampen dendritic excitability, but its largest impact is on the subthreshold range of membrane potentials where the integration of inhibitory and excitatory synaptic inputs takes place. PMID- 9742134 TI - Cloning and expression of two related connexins from the perch retina define a distinct subgroup of the connexin family. AB - We have cloned cDNAs for two closely related connexins (Cx), Cx35 and Cx34.7, from a perch retinal cDNA library. Sequencing of PCR products from genomic DNA revealed that both connexins have an intron 71 bp after the translation initiation site; in Cx35, the intron is 900 bp in length, whereas in Cx34.7 it is approximately 20 kb. Southern blots of genomic DNA suggest that the two connexins represent independent single copy genes. In Northern blots, Cx35 and Cx34.7 transcripts were detected in retina and brain; Cx34.7 also showed a weak signal in smooth muscle (gut) RNA. Antibodies against Cx35 labeled a 30 kDa band on a Western blot of retinal membranes, and in histological sections, the pattern of antibody recognition was consistent with labeling of bipolar cells and unidentified processes in the inner plexiform and nerve fiber layers. When expressed in Xenopus oocytes, Cx35 and Cx34.7 formed homotypic gap junctions, but the junctional conductance between paired oocytes expressing Cx35 was 10-fold greater than that recorded for gap junctional channels formed by Cx34.7. The homotypic gap-junctional channels were closed in a voltage-dependent manner but with relatively weak voltage sensitivity. Heterotypic gap junctions formed by Cx35 and Cx34.7 displayed junctional conductances similar to those of Cx34.7 homotypic pairs and showed a slightly asymmetric current-voltage relationship; the side expressing Cx35 exhibited a higher sensitivity to transjunctional potentials. An analysis of the sequence and gene structure of the connexin family revealed that perch Cx35 and Cx34.7, skate Cx35, and mouse Cx36 constitute a novel gamma subgroup. PMID- 9742135 TI - Tyrosine hydroxylase expression in primary cultures of olfactory bulb: role of L type calcium channels. AB - Sensory activity mediates regulation of tyrosine hydroxylase (TH), the first enzyme in the dopamine biosynthetic pathway, in the rodent olfactory bulb. The current studies established for the first time primary cultures of neonatal mouse olfactory bulb expressing TH and tested whether L-type calcium channels mediate the activity-dependent regulation of the dopamine phenotype. After 1 d in vitro (DIV), a small population of TH-immunostained neurons that lacked extensive processes could be demonstrated. After an additional 2 DIV in serum-free medium, the number of TH neurons had doubled, and they exhibited long interdigitating processes. Membrane depolarization for 48 hr with 50 mM KCl produced a further 2.4-fold increase in the number of TH-immunoreactive neurons compared with control cultures. Increased TH neuron number required at least 36 hr of exposure to KCl. Forskolin, which increases intracellular cAMP levels, induced a 1.5- to 1.6-fold increase in the number of TH-immunostained neurons. Combined treatment with KCl and forskolin was not additive. Nifedipine, an L-type calcium channel blocker, completely prevented the depolarization-mediated increase in TH expression but did not block the response to forskolin. Treatment with Bay K8644, an L-type calcium channel agonist, also significantly increased the number of TH expressing neurons. Depolarization also induced alterations in neuritic outgrowth, resulting in a stellate versus an elongate morphology that, in contrast, was not prevented by nifedipine. These results are the first demonstration that in vitro, as in vivo, depolarization increases TH expression in olfactory bulb and that L-type calcium channels mediate this activity dependent regulation of the dopamine phenotype. PMID- 9742136 TI - Macroscopic and microscopic properties of a cloned glutamate transporter/chloride channel. AB - The behavior of a Cl- channel associated with a glutamate transporter was studied using intracellular and patch recording techniques in Xenopus oocytes injected with human EAAT1 cRNA. Channels could be activated by application of glutamate to either face of excised membrane patches. The channel exhibited strong selectivity for amphipathic anions and had a minimum pore diameter of approximately 5A. Glutamate flux exhibited a much greater temperature dependence than Cl- flux. Stationary and nonstationary noise analysis was consistent with a sub-femtosiemen Cl- conductance and a maximum channel Po << 1. The glutamate binding rate was similar to estimates for receptor binding. After glutamate binding, channels activated rapidly followed by a relaxation phase. Differences in the macroscopic kinetics of channels activated by concentration jumps of L-glutamate or D aspartate were correlated with differences in uptake kinetics, indicating a close correspondence of channel gating to state transitions in the transporter cycle. PMID- 9742137 TI - Morphologically docked synaptic vesicles are reduced in synaptotagmin mutants of Drosophila. AB - Nerve terminal specializations include mechanisms for maintaining a subpopulation of vesicles in a docked, fusion-ready state. We have investigated the relationship between synaptotagmin and the number of morphologically docked vesicles by an electron microscopic analysis of Drosophila synaptotagmin (syt) mutants. The overall number of synaptic vesicles in a terminal was reduced, although each active zone continued to have a cluster of vesicles in its vicinity. In addition, there was an increase in the number of large vesicles near synapses. Examining the clusters, we found that the pool of synaptic vesicles immediately adjacent to the presynaptic membrane, the pool that includes the docked population, was reduced to 24 +/- 5% (means +/- SEM) of control in the sytnull mutation. To separate contributions of overall vesicle depletion and increased spontaneous release from direct effects of synaptotagmin on morphological docking, we examined syt mutants in an altered genetic background. Recombining syt alleles onto a second chromosome bearing an as yet uncharacterized mutation resulted in the expected decrease in evoked release but suppressed the increase in spontaneous release frequency. Motor nerve terminals in this genotype contained more synaptic vesicles than control, yet the number of vesicles immediately adjacent to the presynaptic membrane near active zones was still reduced (33 +/- 4% of control). Our findings demonstrate that there is a decrease in the number of morphologically docked vesicles seen in syt mutants. The decreases in docking and evoked release are independent of the increase in spontaneous release. These results support the hypothesis that synaptotagmin stabilizes the docked state. PMID- 9742139 TI - Whole-cell and single-channel analysis of P-type calcium currents in cerebellar Purkinje cells of leaner mutant mice. AB - The leaner (tgla) mutation in mice results in severe ataxia and an overt neurodegeneration of the cerebellum. Positional cloning has revealed that the tgla mutation occurs in a gene encoding the voltage-activated calcium channel alpha1A subunit. The alpha1A subunit is highly expressed in the cerebellum and is thought to be the pore-forming subunit of P- and Q-type calcium channels. In this study we used both whole-cell and single-channel patch-clamp recordings to examine the functional consequences of the tgla mutation on P-type calcium currents. High-voltage-activated (HVA) calcium currents were recorded from acutely dissociated cerebellar Purkinje cells of homozygous leaner (tgla/tgla) and age-matched wild-type (+/+) mice. In whole cell recordings, we observed a marked reduction of peak current density in tgla/tgla Purkinje cells (-35.0 +/- 1.8 pA/pF) relative to that in +/+ (-103.1 +/- 5.9 pA/pF). The reduced whole-cell current in tgla/tgla cells was accompanied by little to no alteration in the voltage dependence of channel gating. In both genotypes, HVA currents were predominantly of the omega-agatoxin-IVA-sensitive P-type. Cell-attached patch clamp recordings revealed no differences in single-channel conductance between the two genotypes and confirmed the presence of three distinct conductance levels (9, 13-14, and 17-18 pS) in cerebellar Purkinje cells. Analysis of patch open probability (NPo) revealed a threefold reduction in the open-probability of channels in tgla/tgla patches (0.04 +/- 0.01) relative to that in +/+ (0.13 +/- 0.02), which may account for the reduced whole-cell current in tgla/tgla Purkinje cells. These results suggest that the tgla mutation can alter native P-type calcium channels at the single-channel level and that these alterations may contribute to the neuropathology of the leaner phenotype. PMID- 9742138 TI - The cellular and subcellular localization of huntingtin-associated protein 1 (HAP1): comparison with huntingtin in rat and human. AB - The cellular and subcellular distribution of HAP1 was examined in rat brain by light and electron microscopic immunocytochemistry and subcellular fractionation. HAP1 localization was also determined in human postmortem tissue from control and Huntington's disease (HD) cases by light microscopic immunocytochemistry. At the cellular level, the heterogeneity of HAP1 expression was similar to that of huntingtin; however, HAP1 immunoreactivity was more widespread. The subcellular distribution of HAP1 was examined using immunogold electron microscopy. Like huntingtin, HAP1 is a cytoplasmic protein that associates with microtubules and many types of membranous organelles, including mitochondria, endoplasmic reticulum, tubulovesicles, endosomal and lysosomal organelles, and synaptic vesicles. A quantitative comparison of the organelle associations of HAP1 and huntingtin showed them to be almost identical. Within HAP1-immunoreactive neurons in rat and human brain, populations of large and small immunoreactive puncta were visible by light microscopy. The large puncta, which were especially evident in the ventral forebrain, were intensely HAP1 immunoreactive. Electron microscopic analysis revealed them to be a type of nucleolus-like body, which has been named a stigmoid body, that may play a role in protein synthesis. The small puncta, less intensely labeled, were primarily mitochondria. These results indicate that the localization of HAP1 and huntingtin is more similar than previously appreciated and provide further evidence that HAP1 and huntingtin have localizations consistent with roles in intracellular transport. Our data also suggest, however, that HAP1 is not present in the abnormal intranuclear and neuritic aggregates containing the N-terminal fragment of mutant huntingtin that are found in HD brains. PMID- 9742140 TI - Targeting of IgMkappa antibodies to oligodendrocytes promotes CNS remyelination. AB - We previously identified the remyelinating activity of a natural IgMkappa oligodendrocyte-reactive autoantibody (SCH94.03), using a virus-induced murine model of multiple sclerosis. We now describe a second mouse IgMkappa monoclonal antibody (mAb) (SCH79.08) raised against normal mouse spinal cord homogenate, which reacts with myelin basic protein and also promotes remyelination. Because these two mAbs recognize different oligodendrocyte antigens, several previously identified oligodendrocyte-reactive IgMkappa mAbs (O1, O4, A2B5, and HNK-1), each with distinct antigen specificities, were evaluated and found to promote remyelination. In contrast, IgMkappa mAbs that did not bind to oligodendrocytes showed no remyelination. One of these, CH12 IgMkappa mAb, which shares variable region cDNA sequences with SCH94.03 except for amino acid differences in the complementarity-determining region 3 in both heavy and light chains, did not bind to oligodendrocytes and did not promote remyelination. The fact that multiple oligodendrocyte-reactive antibodies with distinct antigen reactivities induce remyelination argues against direct activation by a unique cell surface receptor. These findings are most consistent with the hypothesis that the binding of mAbs to oligodendrocytes in the lesions induces myelin repair via indirect immune effector mechanisms initiated by the mu-chain. Importantly, these studies indicate that oligodendrocyte-reactive natural autoantibodies may provide a powerful and novel therapeutic means to induce remyelination in multiple sclerosis patients. PMID- 9742141 TI - Glial contribution to glutamate uptake at Schaffer collateral-commissural synapses in the hippocampus. AB - Astrocytes in the hippocampus express high-affinity glutamate transporters that are important for lowering the concentration of extracellular glutamate after release at excitatory synapses. These transporters exhibit a permeability to chaotropic anions that is associated with transport, allowing their activity to be monitored in cell-fee patches when highly permeant anions are present. Astrocyte glutamate transporters are highly temperature sensitive, because L glutamate-activated, anion-potentiated transporter currents in outside-out patches from these cells exhibited larger amplitudes and faster kinetics at 36 degreesC than at 24 degreesC. The cycling rate of these transporters was estimated by using paired applications of either L-glutamate or D-aspartate to measure the time necessary for the peak of the transporter current to recover from the steady-state level. Transporter currents in patches recovered with a time constant of 11.6 msec at 36 degreesC, suggesting that either the turnover rate of native transporters is much faster than previously reported for expressed EAAT2 transporters or the efficiency of these transporters is very low. Synaptically activated transporter currents persisted in astrocytes at physiological temperatures, although no evidence of these currents was found in CA1 pyramidal neurons in response to afferent stimulation. L-glutamate-gated transporter currents were also not detected in outside-out patches from pyramidal neurons. These results are consistent with the hypothesis that astrocyte transporters are responsible for taking up the majority of glutamate released at Schaffer collateral-commissural synapses in the hippocampus. PMID- 9742142 TI - A role for cyclin-dependent kinase(s) in the modulation of fast anterograde axonal transport: effects defined by olomoucine and the APC tumor suppressor protein. AB - Proteins that interact with both cytoskeletal and membrane components are candidates to modulate membrane trafficking. The tumor suppressor proteins neurofibromin (NF1) and adenomatous polyposis coli (APC) both bind to microtubules and interact with membrane-associated proteins. The effects of recombinant NF1 and APC fragments on vesicle motility were evaluated by measuring fast axonal transport along microtubules in axoplasm from squid giant axons. APC4 (amino acids 1034-2844) reduced only anterograde movements, whereas APC2 (aa 1034 2130) or APC3 (aa 2130-2844) reduced both anterograde and retrograde transport. NF1 had no effect on organelle movement in either direction. Because APC contains multiple cyclin-dependent kinase (CDK) consensus phosphorylation motifs, the kinase inhibitor olomoucine was examined. At concentrations in which olomoucine is specific for cyclin-dependent kinases (5 microM), it reduced only anterograde transport, whereas anterograde and retrograde movement were both affected at concentrations at which other kinases are inhibited as well (50 microM). Both anterograde and retrograde transport also were inhibited by histone H1 and KSPXK peptides, substrates for proline-directed kinases, including CDKs. Our data suggest that CDK-like axonal kinases modulate fast anterograde transport and that other axonal kinases may be involved in modulating retrograde transport. The specific effect of APC4 on anterograde transport suggests a model in which the binding of APC to microtubules may limit the activity of axonal CDK kinase or kinases in restricted domains, thereby affecting organelle transport. PMID- 9742143 TI - Rapid Ca2+ entry through Ca2+-permeable AMPA/Kainate channels triggers marked intracellular Ca2+ rises and consequent oxygen radical production. AB - The widespread neuronal injury that results after brief activation of highly Ca2+ permeable NMDA channels may, in large part, reflect mitochondrial Ca2+ overload and the consequent production of injurious oxygen radicals. In contrast, AMPA/kainate receptor activation generally causes slower toxicity, and most studies have not found evidence of comparable oxygen radical production. Subsets of central neurons, composed mainly of GABAergic inhibitory interneurons, express AMPA/kainate channels that are directly permeable to Ca2+ ions. Microfluorometric techniques were performed by using the oxidation-sensitive dye hydroethidine (HEt) to determine whether the relatively rapid Ca2+ flux through AMPA/kainate channels expressed on GABAergic neurons results in oxygen radical production comparable to that triggered by NMDA. Consistent with previous studies, NMDA exposures triggered increases in fluorescence in most cultured cortical neurons, whereas high K+ (50 mM) exposures (causing depolarization-induced Ca2+ influx through voltage-sensitive Ca2+ channels) caused little fluorescence change. In contrast, kainate exposure caused fluorescence increases in a distinct subpopulation of neurons; immunostaining for glutamate decarboxylase revealed the responding neurons to constitute mainly the GABAergic population. The effect of NMDA, kainate, and high K+ exposures on oxygen radical production paralleled the effect of these exposures on intracellular Ca2+ levels when they were monitored with the low-affinity Ca2+-sensitive dye fura-2FF, but not with the high-affinity dye fura-2. Inhibition of mitochondrial electron transport with CN- or rotenone almost completely blocked kainate-triggered oxygen radical production. Furthermore, antioxidants attenuated neuronal injury resulting from brief exposures of NMDA or kainate. Thus, as with NMDA receptor activation, rapid Ca2+ influx through Ca2+-permeable AMPA/kainate channels also may result in mitochondrial Ca2+ overload and consequent injurious oxygen radical production. PMID- 9742145 TI - Expression and endocytosis of lysosomal aspartylglucosaminidase in mouse primary neurons. AB - Aspartylglucosaminuria (AGU) is a neurodegenerative lysosomal storage disease that is caused by mutations in the gene encoding for a soluble hydrolase, aspartylglucosaminidase (AGA). In this study, we have used our recently developed mouse model for AGU and analyzed processing, intracellular localization, and endocytosis of recombinant AGA in telencephalic AGU mouse neurons in vitro. The processing steps of AGA were found to be similar to the peripheral cells, but both the accumulation of the inactive precursor molecule and delayed lysosomal processing of the enzyme were detected. AGA was distributed to the cell soma and neuronal processes but was not found in the nerve terminals. Endocytotic capability of cultured telencephalic neurons was comparable to that of fibroblasts, and endocytosis of AGA was blocked by free mannose-6-phosphate (M6P), indicating that uptake of the enzyme was mediated by M6P receptors (M6PRs). Uptake of extracellular AGA was also studied in the tumor-derived cell lines rat pheochromocytoma (PC12) and mouse neuroblastoma cells (N18), which both endocytosed AGA poorly as compared with cultured primary neurons. Expression of cation-independent M6PRs (CI-M6PRs) in different cell lines correlated well with the endocytotic capability of these cells. Although a punctate expression pattern of CI-M6PRs was found in fibroblasts and cultured primary neurons, the expression was beyond the detection limit in PC12 and N18 cells. This indicates that PC12 and N18 are not feasible cell lines to describe neuronal uptake of mannose-6 phosphate-tagged proteins. This in vitro data will form an important basis for the brain-targeted therapy of AGU. PMID- 9742144 TI - Amino acid residues that control pH modulation of transport-associated current in mammalian serotonin transporters. AB - The rat and human serotonin transporters (rSERT and hSERT, respectively) were expressed in Xenopus oocytes and studied using site-directed mutagenesis, electrophysiological recordings, and [3H]5-HT uptake measurements. rSERT, but not hSERT, displayed increased transport-associated current at low pH. Chimeras and point mutations showed that, of the 52 nonidentical residues, a single residue at position 490 (threonine in rSERT and lysine in hSERT) governs this difference. Furthermore, potentiation required the glutamate residue at position 493. Cysteine substitution and alkylation experiments showed that residue 493 is extracellular. Cysteine at 493 increased, whereas aspartate decreased, the net charge movement per transported 5-HT molecule. The mutations at this region did not significantly affect other aspects of SERT function, including agonist independent leakage current, voltage-dependent transient current, and H+ current. This region may therefore be part of an external gate required for rSERT function. The data and analyses show that, in the absence of detailed structural information, a gate-lumen-gate scheme is useful for interpreting results from mutations that alter functional properties of neurotransmitter transporters. PMID- 9742146 TI - The neuronal growth-associated protein GAP-43 interacts with rabaptin-5 and participates in endocytosis. AB - Structural plasticity of nerve cells is a requirement for activity-dependent changes in the brain. The growth-associated protein GAP-43 is thought to be one determinant of such plasticity, although the molecular mechanism by which it mediates dynamic structural alterations at the synapse is not known. GAP-43 is bound by calmodulin when Ca2+ levels are low, and releases the calmodulin when Ca2+ levels rise, suggesting that calmodulin may act as a negative regulator of GAP-43 during periods of low activity in the neurons. To identify the function of GAP-43 during activity-dependent increases in Ca2+ levels, when it is not bound to calmodulin, we sought proteins with which GAP-43 interacts in the presence of Ca2+. We show here that rabaptin-5, an effector of the small GTPase Rab5 that mediates membrane fusion in endocytosis, is one such protein. We demonstrate that GAP-43 regulates endocytosis and synaptic vesicle recycling. Modulation of endocytosis by GAP-43, in association with rabaptin-5, may constitute a common molecular mechanism by which GAP-43 regulates membrane dynamics during its known roles in activity-dependent neurotransmitter release and neurite outgrowth. PMID- 9742147 TI - Increased neurogenesis in the dentate gyrus after transient global ischemia in gerbils. AB - Neurogenesis in the dentate gyrus of adult rodents is regulated by NMDA receptors, adrenal steroids, environmental stimuli, and seizures. To determine whether ischemia affects neurogenesis, newly divided cells in the dentate gyrus were examined after transient global ischemia in adult gerbils. 5-Bromo-2' deoxyuridine-5'-monophosphate (BrdU) immunohistochemistry demonstrated a 12-fold increase in cell birth in the dentate subgranular zone 1-2 weeks after 10 min bilateral common carotid artery occlusions. Two minutes of ischemia did not significantly increase BrdU incorporation. Confocal microscopy demonstrated that BrdU immunoreactive cells in the granule cell layer colocalized with neuron specific markers for neuronal nuclear antigen, microtubule-associated protein-2, and calbindin D28k, indicating that the newly divided cells migrated from the subgranular zone into the granule cell layer and matured into neurons. Newborn cells with a neuronal phenotype were first seen 26 d after ischemia, survived for at least 7 months, were located only in the granule cell layer, and comprised approximately 60% of BrdU-labeled cells in the granule cell layer 6 weeks after ischemia. The increased neurogenesis was not attributable to entorhinal cortical lesions, because no cell loss was detected in this region. Ischemic preconditioning for 2 min, which protects CA1 neurons against subsequent ischemic damage, did not prevent increased neurogenesis in the granule cell layer after a subsequent severe ischemic challenge. Thus, ischemia-induced dentate neurogenesis is not attributable to CA1 neuronal loss. Enhanced neurogenesis in the dentate gyrus may be a compensatory adaptive response to ischemia-associated injury and could promote functional recovery after ischemic hippocampal injury. PMID- 9742148 TI - Regional and cellular patterns of reelin mRNA expression in the forebrain of the developing and adult mouse. AB - The reelin gene encodes an extracellular protein that is crucial for neuronal migration in laminated brain regions. To gain insights into the functions of Reelin, we performed high-resolution in situ hybridization analyses to determine the pattern of reelin expression in the developing forebrain of the mouse. We also performed double-labeling studies with several markers, including calcium binding proteins, GAD65/67, and neuropeptides, to characterize the neuronal subsets that express reelin transcripts. reelin expression was detected at embryonic day 10 and later in the forebrain, with a distribution that is consistent with the prosomeric model of forebrain regionalization. In the diencephalon, expression was restricted to transverse and longitudinal domains that delineated boundaries between neuromeres. During embryogenesis, reelin was detected in the cerebral cortex in Cajal-Retzius cells but not in the GABAergic neurons of layer I. At prenatal stages, reelin was also expressed in the olfactory bulb, and striatum and in restricted nuclei in the ventral telencephalon, hypothalamus, thalamus, and pretectum. At postnatal stages, reelin transcripts gradually disappeared from Cajal-Retzius cells, at the same time as they appeared in subsets of GABAergic neurons distributed throughout neocortical and hippocampal layers. In other telencephalic and diencephalic regions, reelin expression decreased steadily during the postnatal period. In the adult, there was prominent expression in the olfactory bulb and cerebral cortex, where it was restricted to subsets of GABAergic interneurons that co-expressed calbindin, calretinin, neuropeptide Y, and somatostatin. This complex pattern of cellular and regional expression is consistent with Reelin having multiple roles in brain development and adult brain function. PMID- 9742149 TI - Requirement for early-generated neurons recognized by monoclonal antibody lot1 in the formation of lateral olfactory tract. AB - During development, mitral cells, the main output neurons of the olfactory bulb, project axons into a very narrow part of the telencephalon and form an axonal bundle called the lateral olfactory tract (LOT). The present study shows that before the first mitral cell axons elongate, the LOT position is already marked with a subset of early-generated neurons that are recognized by monoclonal antibody lot1 (lot cells). Mitral cell axons choose the lot cell position for their growth pathway and maintain a close contact with the cells until LOT formation is completed. Ablation of lot cells prevented LOT formation in organotypic culture. These results suggest that lot cells are "guidepost cells" for mitral cell axons. PMID- 9742150 TI - Hair cells and supporting cells share a common progenitor in the avian inner ear. AB - Sensory organs of the vertebrate inner ear contain two major cell types: hair cells (HCs) and supporting cells (SCs). To study the lineage relationships between these two populations, replication-defective retroviral vectors encoding marker genes were delivered to the otic vesicle of the chicken embryo. The resulting labeled clones were analyzed in the hearing organ of the chicken, called the basilar papilla (BP), after cellular differentiation. BPs were allowed to develop for 2 weeks after delivery of the retrovirus, were removed, and were processed histochemically as whole mounts to identify clones of cells. Clusters of labeled cells were evident in the sensory epithelium, the nonsensory epithelium, and in adjacent tissues. Labeled cell types included HCs, two morphologically distinct types of SCs, homogene cells, border cells, hyaline cells, ganglion cells, and connective tissue cells. Each clone was sectioned and cell-type identification was performed on sensory clones expressing retrovirally transduced beta-galactosidase. Cell composition was determined for 41 sensory clones, most of which contained both HCs and SCs. Clones containing one HC and one SC were observed, suggesting that a common progenitor exists that can remain bipotential up to its final mitotic division. The possibility that these two cell types may also arise from a mitotic precursor during HC regeneration in the mature basilar papilla is consistent with their developmental history. PMID- 9742152 TI - Glutamate receptor activity is required for normal development of tectal cell dendrites in vivo. AB - Glutamatergic retinotectal inputs mediated principally by NMDA receptors can be recorded from optic tectal neurons early during their morphological development in Xenopus tadpoles. As tectal cell dendrites elaborate, retinotectal synaptic responses acquire an AMPA receptor-mediated synaptic component, in addition to the NMDA component. Here, we tested whether glutamatergic activity was required for the elaboration of dendritic arbors in Xenopus optic tectal neurons. In vivo time-lapse imaging of single DiI-labeled neurons shows that the NMDA receptor antagonist APV (100 microM) blocked the early development of the tectal cell dendritic arbor, whereas the AMPA receptor antagonist CNQX (20 microM) or the sodium channel blocker TTX (1 microM) did not. The decreased dendritic development is attributable to failure to add new branches and extend preexisting branches. These observations indicate that NMDA-type glutamatergic activity promotes the initial development of the dendritic arbor. At later stages of tectal neuron development when AMPA receptor-mediated synaptic transmission is strong, both APV and CNQX decrease dendritic arbor branch length, consistent with a role for glutamatergic synaptic transmission in maintaining dendritic arbor structure. These results indicate that AMPA and NMDA receptors can differentially influence dendritic growth at different stages of neuronal development, in correlation with changes in the relative contribution of the receptor subtype to synaptic transmission. PMID- 9742151 TI - Expression of the mitotic motor protein Eg5 in postmitotic neurons: implications for neuronal development. AB - It is well established that the microtubules of the mitotic spindle are organized by a variety of motor proteins, and it appears that the same motors or closely related variants organize microtubules in the postmitotic neuron. Specifically, cytoplasmic dynein and the kinesin-related motor known as CHO1/MKLP1 are used within the mitotic spindle, and recent studies suggest that they are also essential for the establishment of the axonal and dendritic microtubule arrays of the neuron. Other motors are required to tightly regulate microtubule behaviors in the mitotic spindle, and it is attractive to speculate that these motors might also help to regulate microtubule behaviors in the neuron. Here we show that a homolog of the mitotic kinesin-related motor known as Eg5 continues to be expressed in rodent neurons well after their terminal mitotic division. In neurons, Eg5 is directly associated with the microtubule array and is enriched within the distal regions of developing processes. This distal enrichment is transient, and typically lost after a process has been clearly defined as an axon or a dendrite. Strong expression can resume later in development, and if so, the protein concentrates within newly forming sprouts at the distal tips of dendrites. We suggest that Eg5 generates forces that help to regulate microtubule behaviors within the distal tips of developing axons and dendrites. PMID- 9742153 TI - Genetic analysis on the role of integrin during axon guidance in Drosophila. AB - Heterodimeric cell surface receptor integrin is widely expressed in the nervous system, but its specific role during axon development has not been directly tested in vivo. We show that the Drosophila nervous system expresses low levels of positron-specific (PS) integrin subunits alphaPS1, alphaPS2, and betaPS during embryonic axogenesis. Furthermore, certain subsets of neurons express higher levels of integrin mRNAs than do the rest. Null mutations in either the alphaPS1 or alphaPS2 subunit gene cause widespread axon pathfinding errors that can be rescued by supplying the wild-type integrin subunit to the mutant nervous system. In contrast, misexpressing either the alphaPS1 or alphaPS2 integrin subunit in all neurons leads to no obvious axon pathfinding errors. We propose that integrin does not itself serve as either a "clutch" constituting molecule or a specific growth cone "receptor," as proposed previously, but rather as part of a molecular network that cooperatively guarantees accurate axon guidance. PMID- 9742155 TI - Fibroblast growth factor 2 (FGF-2) promotes acquisition of epidermal growth factor (EGF) responsiveness in mouse striatal precursor cells: identification of neural precursors responding to both EGF and FGF-2. AB - Epidermal growth factor (EGF) and fibroblast growth factor 2 (FGF-2) induce the proliferation of neural precursor cells isolated from specific regions of the embryonic and adult brain. However, the lineage relationship between the EGF- and FGF-2-responsive cells is unknown. In this study we used phosphorylation of the transcription factor cAMP response element-binding protein as a functional readout to identify cells responding to EGF and FGF-2. In primary cultures of mouse embryonic day 14 (E14) striatum, maintained in vitro for 24 hr, 12% of the cells responded to FGF-2, whereas no response to EGF could be detected. Seventy five percent of these FGF-2-responsive cells were beta tubulin III (TuJ1) positive neurons, and 25% expressed nestin, a marker for neuroepithelial precursors. After growth factor treatment for 6 d, a population of nestin positive cells responding to both EGF and FGF-2 were identified. The 6-d-old cultures also contained a small number of TuJ1-positive cells that responded to FGF-2 only. Priming of striatal cells for 24 hr with FGF-2 but not with EGF was sufficient to induce the appearance of EGF- and FGF-2 responsive cells after only 2 d in vitro. Thus, neural precursor cells from the mouse E14 striatum initially responding to FGF-2 only acquire EGF responsiveness later during in vitro development. At this stage EGF and FGF-2 act on the same cells. The acquisition of EGF responsiveness is promoted by FGF-2. PMID- 9742154 TI - Spinal cord neuronal precursors generate multiple neuronal phenotypes in culture. AB - Neuronal restricted precursors (NRPs) () can generate multiple neurotransmitter phenotypes during maturation in culture. Undifferentiated E-NCAM+ (embryonic neural cell adhesion molecule) immunoreactive NRPs are mitotically active and electrically immature, and they express only a subset of neuronal markers. Fully mature cells are postmitotic, process-bearing cells that are neurofilament-M and synaptophysin immunoreactive, and they synthesize and respond to different subsets of neurotransmitter molecules. Mature neurons that synthesize and respond to glycine, glutamate, GABA, dopamine, and acetylcholine can be identified by immunocytochemistry, RT-PCR, and calcium imaging in mass cultures. Individual NRPs also generate heterogeneous progeny as assessed by neurotransmitter response and synthesis, demonstrating the multipotent nature of the precursor cells. Differentiation can be modulated by sonic hedgehog (Shh) and bone morphogenetic protein (BMP)-2/4 molecules. Shh acts as a mitogen and inhibits differentiation (including cholinergic differentiation). BMP-2 and BMP-4, in contrast, inhibit cell division and promote differentiation (including cholinergic differentiation). Thus, a single neuronal precursor cell can differentiate into multiple classes of neurons, and this differentiation can be modulated by environmental signals. PMID- 9742156 TI - Basic FGF increases communication between cells of the developing neocortex. AB - We have found that basic fibroblast growth factor (bFGF), applied to cortical progenitor cells in vitro, produces an increase in the expression of the gap junction protein connexin (Cx) 43 and in the mRNA encoding Cx 43. This effect was evident in both proliferating and nonproliferating cells. The elevated levels of mRNA suggest that bFGF is likely to exert its effect by upregulating the rate of transcription of the Cx 43 gene. We have further shown that the increase in Cx 43 expression is mediated through the receptor tyrosine kinase pathway and is associated with enhanced intercellular dye-coupling mediated by gap junctions. These results suggest that gap junction channels provide a direct conduit for mitogens released in response to bFGF to effectively regulate proliferation during corticogenesis. PMID- 9742157 TI - Promyelinating Schwann cells express Tst-1/SCIP/Oct-6. AB - Tst-1/SCIP/Oct-6, a POU domain transcription factor, is transiently expressed by developing Schwann cells and is required for their normal development into a myelinating phenotype. In tst-1/scip/oct-6-null sciatic nerves, Schwann cells are transiently arrested at the "promyelinating" stage, when they have a one-to-one relationship with an axon but before they have elaborated a myelin sheath. To determine when Schwann cells express Tst-1/SCIP/Oct-6, we examined beta galactosidase (beta-gal) expression in heterozygous tst-1/scip/oct-6 mice, in which one copy of the tst-1/scip/oct-6 gene has been replaced with the LacZ gene. beta-Gal expression from the LacZ gene seems to parallel Tst-1/SCIP/Oct-6 expression from the endogenous tst-1/scip/oct-6 gene in developing and regenerating sciatic nerves. Furthermore, electron microscopic examination of 5bromo-4-chloro-3-indolyl-beta-D-galactopyranoside- (X-gal) and halogenated indolyl-beta-D-galactoside- (Bluo-gal) stained nerves showed that promyelinating Schwann cells express the highest levels of beta-gal, both in developing and in regenerating nerves. Thus, the expression of beta-gal, a surrogate marker of Tst 1/SCIP/Oct-6, peaks at the same stage of Schwann cell development at which development is arrested in tst-1/scip/oct-6-null mice, indicating that Tst 1/SCIP/Oct-6 has a critical role in promyelinating Schwann cells. PMID- 9742158 TI - Development of survival responsiveness to brain-derived neurotrophic factor, neurotrophin 3 and neurotrophin 4/5, but not to nerve growth factor, in cultured motoneurons from chick embryo spinal cord. AB - During embryonic development, most neuronal populations undergo a process usually referred to as naturally occurring neuronal death. For motoneurons (MTNs) of the lumbar spinal cord of chick embryos, this process takes place in a well defined period of time, between embryonic days 6 and 10 (E6-E10). Neurotrophins (NTs) are the best characterized family of neurotrophic factors and exert their effects through activation of their specific Trk receptors. In vitro and in vivo studies have demonstrated that rodent motoneurons survive in response to BDNF, NT3, and NT4/5. In contrast, the trophic dependencies of chicken motoneurons have been difficult to elucidate, and various apparently conflicting reports have been published. In the present study, we describe how freshly isolated motoneurons from E5.5 chick embryos did not respond to any neurotrophin in vitro. Yet, because motoneurons were maintained alive in culture in the presence of muscle extract, they developed a delayed specific survival response to BDNF, NT3, and NT4/5 that is clearly dose-dependent, reaching saturation at doses of 100 pg/ml. This trophic response correlated with increasing expression of the corresponding functional receptors TrkB and TrkC. Moreover, TrkB receptor is able to become autophosphorylated and to activate classical intracellular signaling pathways such as the extracellular signal-regulated protein kinase when it is stimulated with its cognate ligand BDNF. Therefore, our results reconcile the reported differences between in vivo and in vitro studies on the ability of chicken MTNs to respond to some members of the neurotrophin family of trophic factors. PMID- 9742159 TI - Retrograde regulation of growth-associated gene expression in adult rat Purkinje cells by myelin-associated neurite growth inhibitory proteins. AB - Axon regeneration requires that injured neurons reinitiate long-distance growth and upregulate specific genes. To address the question of whether inhibitory environmental cues along the axon could exert a negative, tonic downregulation of growth-associated genes, we have examined adult rat Purkinje cells, which are endowed with poor regenerative capabilities. First we have compared their response to axotomy with that of neurons of the inferior olive, lateral reticular nucleus, and deep cerebellar nuclei, all of which vigorously regenerate into growth-permissive transplants. These injured neurons upregulate the transcription factors c-Jun and JunD, GAP-43, and NADPH diaphorase. In contrast, most axotomized Purkinje cells fail to express any of these markers, showing that the strength of this response parallels the regenerative potential of the examined neuron populations. However, strong upregulation of the same genes can be induced in Purkinje cells after colchicine injection into the uninjured adult cerebellum, indicating that their expression could be controlled by retrograde signals. To assess whether myelin-associated neurite growth inhibitory proteins contribute to this regulation, we applied the neutralizing antibodies IN-1 against one of the main inhibitory components of central myelin (NI-250) either in vivo or in vitro to organotypic cerebellar cultures. Application of IN-1 antibodies induces the upregulation of c-Jun, JunD, and NADPH diaphorase in Purkinje cells, showing that their expression is suppressed constitutively by myelin-associated neurite growth inhibitors. Thus, the inhibitory activity of the IN-1 antigen on axon growth is not restricted to the control of growth cone motility but also involves a retrograde regulation of gene expression in adult central neurons. PMID- 9742160 TI - Interstitial branches develop from active regions of the axon demarcated by the primary growth cone during pausing behaviors. AB - Interstitial branches arise from the axon shaft, sometimes at great distances behind the primary growth cone. After a waiting period that can last for days after extension of the primary growth cone past the target, branches elongate toward their targets. Delayed interstitial branching is an important but little understood mechanism for target innervation in the developing CNS of vertebrates. One possible mechanism of collateral branch formation is that the axon shaft responds to target-derived signals independent of the primary growth cone. Another possibility is that the primary growth cone recognizes the target and demarcates specific regions of the axon for future branching. To address whether behaviors of the primary growth cone and development of interstitial branches are related, we performed high-resolution time-lapse imaging on dissociated sensorimotor cortical neurons that branch interstitially in vivo. Imaging of entire cortical neurons for periods of days revealed that the primary growth cone pauses in regions in which axon branches later develop. Pausing behaviors involve repeated cycles of collapse, retraction, and extension during which growth cones enlarge and reorganize. Remnants of reorganized growth cones are left behind on the axon shaft as active filopodial or lamellar protrusions, and axon branches subsequently emerge from these active regions of the axon shaft. In this study we propose a new model to account for target innervation in vivo by interstitial branching. Our model suggests that delayed interstitial branching results directly from target recognition by the primary growth cone. PMID- 9742161 TI - Functional change of NMDA receptors related to enhancement of susceptibility to neurotoxicity in the developing pontine nucleus. AB - The developing neurons have been reported to be extremely susceptible to toxicity of NMDA during a restricted developmental period. Pontosubicular neuronal necrosis is a typical type of perinatal human brain lesion and often coexists with other forms of cerebral hypoxic and ischemic injuries. To determine whether functional changes of NMDA receptors related to the susceptibility to NMDA toxicity are involved in developing neurons in the pontine nucleus, we have examined the lesion produced by in vivo direct injection of NMDA into the pontine nucleus of rats at postnatal days 1-30, recorded NMDA-induced whole-cell currents from neurons in the pontine nucleus in the developing rat brainstem slices, and performed in situ hybridization for NMDA receptor subunit mRNAs in the pontine nucleus. The susceptibility to NMDA neurotoxicity peaked near postnatal day 15, and the NMDA-induced currents showed prominent reduction of the voltage-dependent block by Mg2+ near postnatal day 15. The pontine nucleus near postnatal day 15 showed distinct expression of the NMDA receptor subunit NR2C mRNA. These results suggest that the susceptibility to NMDA neurotoxicity that is enhanced in the rat pontine nucleus near postnatal day 15 is mediated by the NMDA receptor channels that are relatively insensitive to Mg2+ and that the reduction in the sensitivity of NMDA receptors to Mg2+ correlates with the expression of the NR2C. We present the possibility that functional changes in the NMDA receptor channels play a crucial role in the occurrence of developmentally specific neuronal injury. PMID- 9742162 TI - Brain-derived neurotrophic factor and basic fibroblast growth factor downregulate NMDA receptor function in cerebellar granule cells. AB - Evidence has accumulated to suggest that the NMDA glutamate receptor subtype plays an important role in neuronal degeneration evoked by hypoxia, ischemia, or trauma. Cerebellar granule cells in culture are vulnerable to NMDA-induced neuronal excitotoxicity. In these cells, brain-derived neurotrophic factor (BDNF) and basic fibroblast growth factor (FGF2) prevent the excitotoxic effect of NMDA. However, little is known about the molecular mechanisms underlying the protective properties of these trophic factors. Using cultured rat cerebellar granule cells, we investigated whether BDNF and FGF2 prevent NMDA toxicity by downregulating NMDA receptor function. Western blot and RNase protection analyses were used to determine the expression of the various NMDA receptor subunits (NR1, NR2A, NR2B, and NR2C) after BDNF or FGF2 treatment. FGF2 and BDNF elicited a time-dependent decrease in the expression of NR2A and NR2C subunits. Because NMDA receptor activation leads to increased intracellular Ca2+ concentration ([Ca2+]i), we studied the effect of the BDNF- and FGF2-induced reduction in NR2A and NR2C synthesis on the NMDA-evoked Ca2+ responses by single-cell fura-2 fluorescence ratio imaging. BDNF and FGF2 reduced the NMDA-mediated [Ca2+]i increase with a time dependency that correlates with their ability to decrease NR2A and NR2C subunit expression, suggesting that these trophic factors also induce a functional downregulation of the NMDA receptor. Because sustained [Ca2+]i is believed to be causally related to neuronal injury, we suggest that BDNF and FGF2 may protect cerebellar granule cells against excitotoxicity by altering the NMDA receptor-Ca2+ signaling via a downregulation of NMDA receptor subunit expression. PMID- 9742164 TI - Active membrane properties and signal encoding in graded potential neurons. AB - We investigated the influence of active membrane properties on the precision by which the stimulus velocity is encoded in the membrane potential of a motion sensitive interneuron in the blowfly. The so-called HS-cells respond to visual motion stimuli with a graded shift in membrane potential. Superimposed on this graded response are small spike-like events. This "mixed" visual response mode can be modified by current injection in two different ways. (1) By ongoing injection of hyperpolarizing current, the spike-like events are turned into full blown action potentials, and (2) by injection of depolarizing current, the spike like events become completely suppressed. The visual response then consists of a graded shift of membrane potential only. As a measure of the fidelity, we calculated the coherence between the motion stimulus and the response of the cell elicited with different electrical manipulations of the cell. We found that the coherence was highest for the cell at rest. Any electrical manipulation resulted in a reduced coherence. This was attributable partly to a lower signal-to-noise ratio and partly to an increased nonlinearity in the response. By applying a threshold operation we transformed the analog membrane response into an all-or none spike train. A comparison between these two ways of signal representation revealed that more information about the stimulus velocity is inherent in the analog membrane potential than in the spike train. PMID- 9742163 TI - Presynaptic and postsynaptic actions and modulation of neuroendocrine neurons by a new hypothalamic peptide, hypocretin/orexin. AB - A new orexigenic peptide called hypocretin (orexin) has recently been described in neurons of the lateral hypothalamus and perifornical area. The medial and lateral hypothalamus have been loosely called satiety and feeding centers of the brain, respectively. Approximately one-third of all medial and lateral hypothalamic neurons tested, but not hippocampal neurons, show a striking nanomolar sensitivity to hypocretin. As studied with calcium digital imaging with fura-2, hypocretin raises cytoplasmic calcium via a mechanism based on G-protein enhancement of calcium influx through plasma membrane channels. The peptide has a potent effect at both presynaptic and postsynaptic receptors. Most synaptic activity in hypothalamic circuits is attributable to axonal release of GABA or glutamate. With whole-cell patch-clamp recording, we show that hypocretin, acting directly at axon terminals, can increase the release of each of these amino acid transmitters. Two hypocretin peptides, hypocretin-1 and hypocretin-2, are coded by a single gene; neurons that respond to one peptide also respond to the other. In addition to its effect on feeding, we find that this peptide also regulates the synaptic activity of physiologically identified neuroendocrine neurons studied in hypothalamic slices containing the arcuate nucleus, suggesting a second function of hypocretin in hormone regulation. The widespread distribution of hypocretin axons, coupled with the strong response to the peptide at both presynaptic and postsynaptic sites, suggests that the peptide probably modulates a variety of hypothalamic regulatory systems and could regulate the axonal input to these regions presynaptically. PMID- 9742165 TI - Specific targeting of ganglion cell sprouts provides an additional mechanism for restoring peripheral motor circuits in pelvic ganglia after spinal nerve damage. AB - The pelvic ganglia contain both sympathetic and parasympathetic neurons and provide an interesting model in which to study the effects of a distributed spinal nerve lesion. Previous animal studies have suggested that after either lumbar or sacral nerve injury, some functional connections are restored between preganglionic and postganglionic neurons. It has been proposed that this is because of intact preganglionic axons sprouting collaterals to supply denervated ganglion cells. However, this has never been demonstrated, and our study has investigated whether the ganglion cells themselves contribute to axogenesis and restoration of peripheral circuitry. We have monitored the growth of axons from pelvic ganglion cells after lumbar or sacral nerve injury (partial decentralization), or a combination of the two (total decentralization). These new processes were distinguished from intact preganglionic terminals by their immunoreactivity for substances present only in pelvic ganglion neurons (vasoactive intestinal peptide, neuropeptide Y, and tyrosine hydroxylase). The proportion of pelvic neurons surrounded by these immunostained fibers was then assessed. Complete removal of preganglionic terminals provides the biggest stimulus for growth of new axon processes (sprouts), which grow profusely within just a few days. These arise from each of the main chemical classes of pelvic neurons but grow at different rates and have different distributions. Importantly, some chemical classes of sprouts preferentially supply neurons of dissimilar histochemistry, suggesting the presence of very specific targeting mechanisms rather than random growth. These sprouts are transient, however, those formed after partial decentralization appear to be maintained. Moreover, after lesion of either lumbar or sacral spinal nerves, many sprouts arise from neurons with intact spinal connections and innervate neurons that have lost their preganglionic inputs. This provides a very different alternative mechanism to reestablish communication between preganglionic and postganglionic neurons. In conclusion, we have demonstrated a rapid and selective axogenesis within the pelvic ganglion after spinal nerve injury. This may allow the development of novel strategies by which autonomic nerve pathways can be experimentally manipulated, to facilitate more rapid return of appropriate peripheral reflex control. PMID- 9742166 TI - Decreased presynaptic sensitivity to adenosine after cocaine withdrawal. AB - The nucleus accumbens (NAc) is a site mediating the rewarding properties of drugs of abuse, such as cocaine, amphetamine, opiates, nicotine, and alcohol (Wise and Bozarth, 1987; Koob, 1992; Samson andHarris, 1992; Woolverton and Johnson, 1992; Self and Nestler, 1995; Pontieri et al., 1996). Acute cocaine has been shown to decrease excitatory synaptic transmission mediated by the cortical afferents to the NAc (Nicola et al., 1996), but the effects of long-term cocaine treatment and withdrawal have not been explored. Here, we report that long-term (1 week) withdrawal from chronic cocaine reduced the potency of adenosine to presynaptically inhibit glutamate (Glu) release by activating adenosine A1 receptors. Adenosine A1 receptors were not desensitized, because the potency of the metabolically stable adenosine analog N6-cyclopentyl-adenosine was unchanged after chronic cocaine withdrawal. When adenosine transporters were blocked, the potency of adenosine to inhibit Glu release from naive and cocaine-withdrawn NAc slices was similar. These results suggest that one of the long-term consequences of cocaine withdrawal is an augmented uptake of adenosine. This long-lasting change expressed at the presynaptic excitatory inputs to the medium spiny output neurons in the NAc may help identify new therapeutic targets for the treatment of drug abuse. PMID- 9742167 TI - Electrophysiological characterization of GABAergic neurons in the ventral tegmental area. AB - GABAergic neurons in the ventral tegmental area (VTA) play a primary role in local inhibition of mesocorticolimbic dopamine (DA) neurons but are not physiologically or anatomically well characterized. We used in vivo extracellular and intracellular recordings in the rat VTA to identify a homogeneous population of neurons that were distinguished from DA neurons by their rapid-firing, nonbursting activity (19.1 +/- 1.4 Hz), short-duration action potentials (310 +/- 10 microseconds), EPSP-dependent spontaneous spikes, and lack of spike accommodation to depolarizing current pulses. These non-DA neurons were activated both antidromically and orthodromically by stimulation of the internal capsule (IC; conduction velocity, 2.4 +/- 0.2 m/sec; refractory period, 0.6 +/- 0.1 msec) and were inhibited by stimulation of the nucleus accumbens septi (NAcc). Their firing rate was moderately reduced, and their IC-driven activity was suppressed by microelectrophoretic application or systemic administration of NMDA receptor antagonists. VTA non-DA neurons were recorded intracellularly and showed relatively depolarized resting membrane potentials (-61.9 +/- 1.8 mV) and small action potentials (68.3 +/- 2.1 mV). They were injected with neurobiotin and shown by light microscopic immunocytochemistry to be multipolar cells and by electron microscopy to contain GABA but not the catecholamine-synthesizing enzyme tyrosine hydroxylase (TH). Neurobiotin-filled dendrites containing GABA received asymmetric excitatory-type synapses from unlabeled terminals and symmetric synapses from terminals that also contained GABA. These findings indicate that VTA non-DA neurons are GABAergic, project to the cortex, and are controlled, in part, by a physiologically relevant NMDA receptor-mediated input from cortical structures and by GABAergic inhibition. PMID- 9742168 TI - Proprioceptive input to feeding motor programs in Aplysia. AB - Although central pattern generators (CPGs) can produce rhythmic activity in isolation, it is now generally accepted that under physiological conditions information from the external and internal environment is incorporated into CPG induced motor programs. Experimentally advantageous invertebrate preparations may be particularly useful for studies that seek to characterize the cellular mechanisms that make this possible. In these experiments, we study sensorimotor integration in the feeding circuitry of the mollusc Aplysia. We show that a premotor neuron with plateau properties, B51, is important for generating the radula closing/retraction phase of ingestive motor programs. When B51 is depolarized in semi-intact preparations, radula closing/retractions are enhanced. When B51 is hyperpolarized, radula closing/retractions are reduced in size. In addition to being important as a premotor interneuron, B51 is also a sensory neuron that is activated when the feeding apparatus, the radula, rotates backward. The number of centripetal spikes in B51 is increased if the resistance to backward rotation is increased. Thus, B51 is a proprioceptor that is likely to be part of a feedback loop that insures that food will be moved into the buccal cavity when difficulty is encountered. Our data suggest, therefore, that Aplysia are able to adjust feeding motor programs to accommodate the specific qualities of the food ingested because at least one of the neurons that generates the basic ingestive motor program also serves as an on-line monitor of the success of radula movements. PMID- 9742169 TI - Melatonin entrains the restored circadian activity rhythms of syrian hamsters bearing fetal suprachiasmatic nucleus grafts. AB - A circadian pacemaker consists of at least three essential features: the ability to generate circadian oscillations, an output signal, and the ability to be entrained by external signals. In rodents, ablation of the suprachiasmatic nucleus (SCN) results in the loss of circadian rhythms in activity. Rhythmicity can be restored by transplanting fetal SCN into the brain of the lesioned animal, demonstrating the first two of the essential pacemaker features within the grafts. External signals, such as the light/dark cycle, have not, however, been shown to entrain the restored rhythms. Melatonin injections are an effective entraining stimulus in fetal and neonatal Syrian hamsters of the same developmental ages used to provide donor tissue for transplantation. Therefore, melatonin was used to test the hypothesis that SCN grafts contain an entrainable pacemaker. Daily injections of melatonin were given to SCN-lesioned hosts beginning on the day after transplantation of fetal SCN. Two groups that received melatonin at different times of day 12 hr apart each showed significantly clustered phases but with average phases that differed by 8.67 hr. Thus melatonin was able to entrain the restored circadian activity rhythms. In contrast to these initial injections, injections given 6 weeks after transplantation were unable to entrain or phase shift the rhythms. The results demonstrate that SCN grafts contain an entrainable circadian pacemaker. In addition, the results also indicate that the fetal SCN is directly sensitive to melatonin and, as with intact hamsters, sensitivity to melatonin is lost during SCN development. PMID- 9742170 TI - Removal of cholinergic input to rat posterior parietal cortex disrupts incremental processing of conditioned stimuli. AB - Recent research suggests that the basal forebrain cholinergic neurons innervating the cortex play a role in attentional functions in both primates and rodents. Among the cortical targets of these projections in primates is the posterior parietal cortex (PPC), a region shown to be critically involved in the regulation of attention. Recent anatomical studies have defined a cortical region in the rat that may be homologous to the PPC of primates. In the present study, cholinergic innervation of the PPC was depleted by intracortical infusion of the immunotoxin 192 IgG-saporin. Control and lesioned rats were then tested in two associative learning paradigms designed to increase attentional processing of conditioned stimuli (CSs). In one experiment, attention was manipulated by shifting a predictive relation between a light CS and another CS to a less predictive relation. Unlike control rats, lesioned rats failed to increase attention when the predictive relation was modified. In a second experiment, attentional processing of a tone CS was increased when its introduction during training coincided with a change in the value of the unconditioned stimulus, a phenomenon referred to as unblocking. Unlike control rats, lesioned rats failed to exhibit unblocking. In both paradigms, lesioned rats conditioned normally when the training procedures did not encourage increased attentional processing. These findings, across different behavioral paradigms and stimulus modalities, provide converging evidence that intact cholinergic innervation of the PPC is important for changes in attention that can increase the processing of certain cues. PMID- 9742171 TI - Long-term dietary strawberry, spinach, or vitamin E supplementation retards the onset of age-related neuronal signal-transduction and cognitive behavioral deficits. AB - Recent research has indicated that increased vulnerability to oxidative stress may be the major factor involved in CNS functional declines in aging and age related neurodegenerative diseases, and that antioxidants, e.g., vitamin E, may ameliorate or prevent these declines. Present studies examined whether long-term feeding of Fischer 344 rats, beginning when the rats were 6 months of age and continuing for 8 months, with diets supplemented with a fruit or vegetable extract identified as being high in antioxidant activity, could prevent the age related induction of receptor-mediated signal transduction deficits that might have a behavioral component. Thus, the following parameters were examined: (1) oxotremorine-enhanced striatal dopamine release (OX-K+-ERDA), (2) cerebellar beta receptor augmentation of GABA responding, (3) striatal synaptosomal 45Ca2+ clearance, (4) carbachol-stimulated GTPase activity, and (5) Morris water maze performance. The rats were given control diets or those supplemented with strawberry extracts (SE), 9.5 gm/kg dried aqueous extract (DAE), spinach (SPN 6.4 gm/kg DAE), or vitamin E (500 IU/kg). Results indicated that SPN-fed rats demonstrated the greatest retardation of age-effects on all parameters except GTPase activity, on which SE had the greatest effect, whereas SE and vitamin E showed significant but equal protection against these age-induced deficits on the other parameters. For example, OX-K+-ERDA enhancement was four times greater in the SPN group than in controls. Thus, phytochemicals present in antioxidant-rich foods such as spinach may be beneficial in retarding functional age-related CNS and cognitive behavioral deficits and, perhaps, may have some benefit in neurodegenerative disease. PMID- 9742173 TI - Duodenal sensory neurons project to sphincter of Oddi ganglia in guinea pig. AB - Retrograde labeling of duodenum-sphincter of Oddi (SO) preparations in vitro with the carbocyanine dye DiI revealed that duodenal neurons project to the SO. The duodenum-SO-projecting neurons were immunoreactive (IR) for choline acetyltransferase but not nitric oxide synthase or calretinin, indicating that this is a cholinergic projection and that this pathway is distinct from the circuitry involved in the ascending limb of the peristaltic reflex. Approximately 20% of the duodenum-SO projection neurons were IR for calbindin. Calbindin-IR nerves within SO ganglia degenerated when the SO was maintained in organ culture alone, but persisted when the SO was cultured with the duodenum intact. Therefore, SO ganglia are a target of the calbindin-positive duodenum-SO projection. Because calbindin is a marker of intrinsic sensory neurons that have processes that pass to the mucosa, these neurons are in position to detect the release of a compound from the mucosa and signal its release to SO ganglia. When applied to retrogradely labeled neurons, cholecystokinin (CCK) elicited a prolonged depolarization, indicating that duodenum-SO-projecting neurons could be capable of detecting CCK released from the mucosa. It is proposed that the role of the intrinsic sensory neurons that project to the SO may be to signal the postprandial release of CCK, thus providing an instruction to decrease SO resistance and facilitate the flow of bile into the duodenum. PMID- 9742172 TI - Evidence that excitatory amino acid receptors within the temporomandibular joint region are involved in the reflex activation of the jaw muscles. AB - We have previously shown that injection of the inflammatory irritant and small fiber excitant mustard oil (MO) into the temporomandibular joint (TMJ) region can reflexively induce a prolonged increase in the activity of both digastric and masseter muscles in rats. It is possible that peripheral excitatory amino acid (EAA) receptors play a role in this effect, because MO-evoked increases in jaw muscle activity are attenuated by preapplication of the noncompetitive NMDA receptor antagonist MK-801 into the TMJ region. In the present study the EAA receptor agonists glutamate, NMDA, kainate, and AMPA were applied locally to the TMJ region. Jaw muscle responses similar to those evoked by MO application to the TMJ region were achieved with glutamate, NMDA, AMPA, and kainate. Repeated application of glutamate, NMDA, or AMPA at intervals of 30 min evoked responses in the ipsilateral jaw muscles that were of comparable magnitude. Co-application of the NMDA receptor antagonist DL-2-amino-5-phosphonovalerate (0.5 micromol) significantly reduced the magnitude of the glutamate- and NMDA-evoked ipsilateral jaw muscle responses without affecting responses evoked by AMPA. In contrast, co application of the non-NMDA receptor antagonist 6-cyano-7-nitroquinoxaline-2,3 dione (1 nmol) significantly reduced the magnitude of the glutamate- and AMPA evoked ipsilateral jaw muscle responses without affecting responses evoked by NMDA. This evidence suggests that both NMDA and non-NMDA EAA receptor types are located within the TMJ region and may contribute to jaw muscle activity that can be reflexively evoked from the TMJ region. PMID- 9742174 TI - Dopamine depletion reorganizes projections from the nucleus accumbens and ventral pallidum that mediate opioid-induced motor activity. AB - Motor activity elicited pharmacologically from the nucleus accumbens by the mu opioid receptor agonist D-Ala-Tyr-Gly-NMePhe-Gly-OH (DAMGO) is augmented in rats sustaining dopamine depletions. GABAergic projections from the nucleus accumbens to ventral pallidum and ventral tegmental area (VTA) are involved because stimulation of GABAB receptors in the VTA (by baclofen) or GABAA receptors in the ventral pallidum (by muscimol) inhibit the motor response induced by the microinjection of DAMGO into the nucleus accumbens. The present study was done to determine which of these projections is mediating the augmented DAMGO-induced motor activity that follows 6-hydroxydopamine lesions of the nucleus accumbens. The inhibition of DAMGO-induced activation by pallidal injections of muscimol was markedly attenuated in lesioned animals, whereas the inhibition by VTA injections with baclofen was greatly enhanced. A similar switch in emphasis from pallidal to mesencephalic efferents was not observed for dopamine-induced motor activity, because muscimol microinjections inhibited the response elicited by dopamine microinjection into the nucleus accumbens in all subjects. The stimulation of mu opioid receptors in the ventral pallidum also elicits motor activation, and this is blocked by baclofen microinjection into the VTA. However, after dopamine depletion in the nucleus accumbens, baclofen in the VTA was ineffective in blocking the motor response by DAMGO in the ventral pallidum. These data reveal that dopamine depletion in the nucleus accumbens produces a lesion-induced plasticity that alters the effect of mu-opioid receptor stimulation on efferent projections from the nucleus accumbens and ventral pallidum. PMID- 9742175 TI - Ascending projections of simple and complex cells in layer 6 of the cat striate cortex. AB - Receptive field properties vary systematically across the different layers of the cat striate cortex. Understanding how these functional differences emerge requires a precise description of the interlaminar connections and the quality of information that they transmit. This study examines the contribution of the two physiological types of neuron in layer 6, simple and complex, to the cortical microcircuit. The approach was to make whole-cell recordings with dye-filled electrodes in vivo to correlate visual response property with intracortical projection pattern. The two simple cells we stained projected to layer 4, as previously reported (Gilbert and Wiesel, 1979; Martin and Whitteridge, 1984). Six of the eight complex cells that we labeled projected to the superficial layers, a pathway not previously described in the cat. The remaining two cells targeted the infragranular layers. Layer 4 is dominated by simple cells, whereas layers 5 and 2+3 are mainly composed of complex cells (Hubel and Wiesel, 1962; Gilbert, 1977). Hence, our results indicate that the ascending projections of simple cells in layer 6 target other simple cells. In parallel, the ascending projections of a population of complex cells in layer 6 favor other complex cells. Anatomical experiments in several species (Lund and Boothe,1975; Burkhalter,1989; Usrey and Fitzpatrick, 1996; Wiser and Callaway, 1996) had also demonstrated that layer 6 gives rise to two separate intracortical pathways. Pooling the results of these anatomical studies with our own suggests a common feature of the laminar organization: cells that project to different intracortical targets have distinct functional characteristics. PMID- 9742177 TI - Mental-attentional capacity: does cognitive style make a difference? AB - There is currently no consensus on whether the difference between field-dependent and field-independent subjects on tasks of cognitive abilities result from different mental processing strategies, from true group differences in cognitive ability, or from both. School-age children (N = 239) were tested for field dependence/independence using the Children's Embedded Figures Test and for mental attentional capacity using the Figural Intersection Task. Multigroup scaling models were used to separate the contributions of style from ability in children's performance on Figural Intersection items. Results show that field dependent children have greater odds of success than field-independent children in Figural Intersection items when the task's mental-attentional demand is above the child's mental attentional capacity, as assessed in the same task. The contrary is true when the task's mental-attentional demand is below or equal to the mental-attentional capacity of the child. Overall, field-dependent children obtain lower estimates of mental-attentional capacity than field-independent children in this task. We discuss the implications of these results for the measurement of mental-attentional capacity and the conceptualization of field dependence/independence. PMID- 9742176 TI - Cellular and synaptic modulation underlying substance P-mediated plasticity of the lamprey locomotor network. AB - The tachykinin substance P modulates the lamprey locomotor network by increasing the frequency of NMDA-evoked ventral root bursts and by making the burst activity more regular. These effects can last in excess of 24 hr. In this paper, the effects of substance P on the synaptic and cellular properties of motor neurons and identified network interneurons have been examined. Substance P potentiated the amplitude of monosynaptic glutamatergic inputs from excitatory interneurons and reticulospinal axons. The amplitude and frequency of miniature EPSPs was increased, suggesting that the synaptic modulation was mediated presynaptically and postsynaptically. The postsynaptic modulation was caused by a specific effect of substance P on the NMDA component of the synaptic input, whereas the presynaptic component was calcium-independent. Substance P did not affect monosynaptic glycinergic inputs from lateral interneurons, crossed inhibitory interneurons, or ipsilateral segmental interneurons or postsynaptic GABAA or GABAB responses, suggesting that it has little effect on inhibitory synaptic transmission. At the cellular level, substance P increased synaptic inputs, resulting in membrane potential oscillations in motor neurons, crossed caudal interneurons, lateral interneurons, and excitatory interneurons. The spiking in response to depolarizing current pulses was increased in motor neurons, lateral interneurons, and excitatory interneurons, but usually was reduced in crossed inhibitory interneurons. Substance P reduced the calcium-dependent afterhyperpolarization after an action potential in motor neurons and lateral interneurons, but did not affect this conductance in excitatory or crossed inhibitory interneurons. The relevance of these cellular and synaptic changes to the modulation of the locomotor network is discussed. PMID- 9742178 TI - Developmental differences in implicit and explicit memory performance. AB - Performance of preschool, elementary school, and college students was compared on a series of perceptual and conceptual implicit and explicit memory tasks that followed perceptual or conceptual processing during study. As expected, performance on the conceptual explicit memory task improved across age groups. In contrast, performance on the perceptual explicit memory task as well as that on both types of implicit memory tasks showed no developmental change. Also, perceptual processing during study led to better memory performance than conceptual processing for both the perceptual implicit and perceptual explicit tasks and conceptual processing during study led to better memory performance on the conceptual explicit memory task. Performance on the conceptual implicit memory task, in contrast, was affected equally by both types of study processing. The results are discussed in terms of transfer-appropriate processing (Roediger & Blaxton, 1987b) and unitization and grouping processes (Graf & Schacter, 1989). PMID- 9742179 TI - Temporal organization in children's strategy formation. AB - It is well established that children use study behaviors such as card sorting, category naming, and item-by-item rehearsal to assist subsequent word recall. In this article, we provide evidence that these behaviors are organized into coherent temporal patterns. Fourier analyses of individual behaviors over a sequence of five consecutive study/recall trials indicated that sorting was synchronized with the start of each trial, whereas rehearsal tended to occur later in each trial. Fourier analyses of pairs of behaviors indicated that sorting and category naming, both concerned with categorization of the to-be remembered words, co-occurred early in each trial at a greater rate than expected based on their individual frequencies of occurrence (i.e., they were used cooperatively). In contrast, verbal rehearsal of individual words co-occurred with both sorting and category naming at a lesser rate than would be expected based on their individual frequencies of occurrence. The results thus point to a global strategy in which children learn the items' categories before they learn them individually. There was little apparent qualitative difference in temporal organization for second- and fourth-grade children. However, sorting early in each trial was more pronounced for children with better word recall (regardless of grade), and the suppressed co-occurrence of rehearsal with sorting and category naming (i.e., keeping category learning temporally separate from item learning) was more pronounced for the fourth-grade children. PMID- 9742180 TI - Perception of three-dimensional cues in early infancy. AB - Three experiments examined infants' processing of 3D information in static images. Three-month-olds familiarized with micropatterns that appeared to adults to be 3D blocks illuminated from the top subsequently preferred a test array that contained a micropattern that appeared to be a block illuminated from the bottom; however, they failed to detect comparable discrepancies in 2D images. Thus, 3 month-olds are sensitive to 3D cues in static images. In another experiment, infants failed to exhibit a preference between an array containing a single novel element among familiar elements and another array containing a single familiar element among novel elements. Thus, unlike discrepancies in fundamental features such as line-crossings, discrepancies in 3D cues failed to pop-out and engage infants' attention. These results reveal that 3-month-olds are sensitive to 3D cues in static images. However, discrepancies based on these cues may not engage infants' attention like those based on fundamental features. PMID- 9742182 TI - Segmentation, not rhyming, predicts early progress in learning to read. AB - We present a longitudinal study of children in the first 2 years of learning to read. A battery of tests of phonological skills administered when the children were prereaders identified two distinct and relatively independent factors, Rhyming (defined by measures of rhyme detection and rhyme production) and Segmentation (defined by measures of phoneme identification and phoneme deletion). Segmentation was strongly correlated with attainment in reading and spelling at the end of the first year at school, while Rhyming was not. In addition, letter name knowledge predicted both reading and spelling skill and showed an interactive effect with children's segmentation skills. By the end of the second year of school, however, rhyming had started to exert a predictive effect on spelling, but not on reading. The results are discussed in the context of current theories of the role of phonological skills in learning to read. PMID- 9742183 TI - Sensitivity to onset and rhyme does predict young Children's reading: a comment on Muter, Hulme, Snowling, and Taylor (1997) AB - Muter, Hulme, Snowling, and Taylor (1997) claimed that measures of phoneme segmentation, and not measures of rhyme, predict young children's reading. They base this claim on the relative predictive success of two rhyme and two phoneme segmentation tasks. However, there is a problem with one of their two rhyme measures, the Rhyme Detection measure. The children were asked to select a choice word which "rhymes with or sounds like" a target word, but the authors only scored rhyme choices ("boat"-"coat") as correct. Choices of words with the same onset as the target ("train"-"tractor") were counted as mistakes, even though these latter choices also shared a common sound with the target. A better way to score the task is to count onset as well as rhyme choices as correct. The new score predicts reading and spelling as well as the phoneme tasks. This result is consistent with the hypothesis of Goswami and Bryant (1990) that sensitivity to onset and rhyme, as well as awareness of phonemes, plays a part in children's success in reading and to spelling. PMID- 9742184 TI - Segmentation does predict early progress in learning to read better than rhyme: A reply to bryant AB - In our recent paper (Muter, Hulme, Snowling, & Taylor, 1997) we argued that measures of segmentation were better predictors of early progress in learning to read than were measures of rhyme. Bryant (1998, this issue), in his comment on our paper, has argued that this conclusion is flawed because the instructions used in our rhyme detection measure included the phrase "rhymes with or sounds like." We present new data showing that the instructions used do not have the effect Bryant claims: asking children which word "rhymes with" or which word "rhymes with or sounds like" a target word produces identical patterns of responses. We argue that Bryant's new measure derived from our data simply reflects children's global sensitivity to the similarity in sound between different words and that this measure provides no convincing support for his conclusion that sensitivity to onset and rime is a predictor of children's success in learning to read. We conclude that the data in our paper, as well as other recent evidence, support the view that measures of phonemic segmentation are better predictors of early reading skills than are measures of onset-rime sensitivity. Copyright 1998 Academic Press. PMID- 9742185 TI - Transfer effects across contextual and linguistic boundaries: evidence from poor readers. AB - We report two experiments that are consistent with two hypotheses about poor, nonfluent readers: (1) fluency gains in text reading skill transfer across contextual and linguistic boundaries and (2) these fluency gains enable higher order comprehension operations to function in the processing of text. We conclude that unlike the fluent reader, the nonfluent reader does not completely integrate the surface characteristics (words) of the text and the message of the text. Word level representations remain free to support transfer across various processing episodes. Thus, a variety of reading experiences aimed at promoting word recognition fluency will provide benefits to the developing reader. PMID- 9742186 TI - Long- and short-looking infants' recognition of symmetrical and asymmetrical forms. AB - Adults process symmetrical visual forms more rapidly than asymmetrical visual forms, presumably because symmetrical forms are amenable to a global visual encoding strategy. Individual differences in look duration during infancy have been hypothesized to covary with different modes of visual intake and encoding, with longer look durations reflecting encoding based on prolonged inspection of local visual properties, and briefer look durations reflecting encoding based on more of a global, or global-to-local processing sequence. This hypothesis predicts that short-looking infants would process symmetrical stimuli faster than asymmetrical stimuli, but that long-looking infants would not. Three experiments are described here in which this prediction is tested. Results were in general accord with the prediction, and provide further support for the hypothesis that individual differences in look duration may reflect different modes of visual encoding or inspection. PMID- 9742187 TI - Fungal genetics and biology. PMID- 9742188 TI - An improved method for affinity probe localization in whole cells of filamentous fungi. AB - The fungal cell wall, though phylogenetically variable, acts universally as a potent barrier to probing intracellular structures. Thus, the use of high molecular-weight probes such as antibodies and lectins has proven a formidable challenge. We have devised a preparative method for use with various affinity probes that can be applied to a broad spectrum of filamentous fungal species and used for imaging whole cells. In this study, confocal imaging of whole-mount fungal hyphae after freeze substitution, methacrylate embedment/de-embedment, and infiltration with affinity probes has yielded remarkably improved renderings of the three-dimensional distribution of both microtubules (using antibodies against both alpha- and beta-tubulin) and concanavalin A binding sites. Using this protocol we have been able to document: (1) the three-dimensional distribution of microtubules in all regions of hyphae, (2) the presence of apparent foci for cytoplasmic microtubules, (3) persistent cytoplasmic microtubules during mitosis, and (4) a three-dimensional view of many compartments of the endomembrane system including Golgi-equivalent organelles and apical vesicles. The last result represents the first direct confirmation of apical vesicles comprising the Spitzenkorper. PMID- 9742189 TI - Calcium imaging: a primer for mycologists. AB - This article aims to encourage more fungal biologists to consider the imaging of cytoplasmic Ca2+ fluxes. Compared to other organisms, for fungi there have been remarkably few attempts to characterize the role of Ca2+ fluxes in signal transduction and general cellular activities, even though other approaches indicate that fungal growth and development are highly dependent upon Ca2+. The methodologies for imaging Ca2+ fluxes continue to develop rapidly. These methodologies are explained here in a style that should be accessible to a newcomer to the field, hopefully forming a bridge to the more complex methodological literature. PMID- 9742190 TI - Dynamic rearrangement of the filamentous actin network occurs during zoosporogenesis and encystment in the oomycete phytophthora cinnamomi AB - The organization of filamentous actin (F-actin) in living cells of the oomycete Phytophthora cinnamomi was determined during zoosporogenesis and zoospore encystment by microinjecting sporangia with fluorescently labeled phalloidin and observing resultant fluorescence by confocal microscopy. In multinucleate sporangia prior to the induction of cleavage, phalloidin labeling took the form of plaques which occurred mainly in the periphery of the sporangia. After induction of cleavage, phalloidin labeling showed that the plaques disappeared and that F-actin began to accumulate along the developing cleavage planes and around nuclei and water expulsion vacuoles. F-actin labeling was also observed near the plasma membrane in zoospores and young cysts but reverted to the plaque form in older cysts. Localization of F-actin close to the developing cleavage planes is consistent with the idea that actin microfilaments function in the positioning and expansion of the cleavage membranes. Observations of plaques of actin in living sporangia provide evidence that plaques are not aldehyde-induced fixation artifacts. Copyright 1998 Academic Press. PMID- 9742191 TI - Establishment and maintenance of nuclear position during zoospore formation in allomyces macrogynus: roles of the cytoskeleton AB - The involvement of the microtubule (MT) and actin microfilament (MF) cytoskeletons in establishing nuclear positions during zoosporogenesis in Allomyces macrogynus was assessed using selective cytoskeletal disrupting treatments and documented with light microscopy. These experiments were coupled with low-speed centrifugation studies to determine the degree to which cytoskeletal elements anchor nuclear position. At the onset of zoospore formation, nuclei were positioned only in cortical cytoplasmic regions of the zoosporangia (ZS). Immunofluorescence microscopy revealed that MTs primarily emanated from centrosomal regions into the surrounding cytoplasm at this stage. During delimitation of the cytoplasm into individual uninucleate zoospores, nuclei migrated from cortical regions to become distributed throughout the cytoplasm. Coincident with nuclear migrations, MTs were primarily organized at and emanated from nuclear surfaces, forming extensive perinuclear arrays. Nuclear migrations were suppressed in ZS induced to sporulate in the presence of cytochalasin D, an actin MF inhibiting compound. Disruption of MTs with nocodazole did not block nuclear migrations, although resultant nuclear spacing was irregular. Centrifugation treatments of control and drug-treated ZS demonstrated that nuclear positions were stabilized by perinuclear MT arrays. The results indicate that nuclear motility in ZS of A. macrogynus is the result of an actin-based system while perinuclear MTs arrays function to establish and fix nuclear position during zoospore formation. Copyright 1998 Academic Press. PMID- 9742192 TI - Injection tube differentiation in gun cells of a haptoglossa species which infects nematodes AB - The gun cells which develop from germinating cysts in Haptoglossa produce a specialized infection apparatus, the injection tube. Upon eversion this tube fires a missile-like projectile which penetrates the host cuticle and then forms an infective sporidium within the body cavity of the nematode host. The temporal assembly of this complex cell organelle has been determined by serial-section reconstructions of maturing gun cells in a previously undescribed Haptoglossa species. The differentiation of the partially walled inverted injection tube is an unusual example of internal tube growth, in which membrane and wall assembly are temporally separated. There is no evidence that the shape of this inverted tube, which coils around the nucleus until it doubles back on itself, is dictated by the disposition of cytoplasmic microtubules. However, actin-like material was associated with the delimiting membrane of the differentiating tube, particularly in the regions of extension. From these studies it seems likely that the "head and buttress" structures previously depicted as the barbed tip of the "harpoon like" penetration missile are part of a separate, structurally complex system which we suggest locks the "missile" into position in the invaginated injection tube. From this detailed account of cell architecture, models for the likely mechanism of infection cell firing are discussed, and unresolved questions relating to the cell biology and biochemistry of these complex organelles are highlighted. Copyright 1998 Academic Press. PMID- 9742193 TI - Mapping fungal ion channel locations AB - Ion channel mapping techniques are described and the results for two fungal organisms, Saprolegnia ferax and Neurospora crassa, are presented. In these species, two channel types have been characterized, stretch-activated channels exhibiting significant calcium permeability and spontaneous channels having significant potassium permeability. Two distinct analyses of patch clamp data, analysis of channel self-clustering and association between different channel types, and localization along the hyphae, reveal significant differences between the two organisms. S. ferax maintains a tip-high gradient of both channel types which is lost after disruption of the actin cytoskeleton. There is significant self-clustering of the channels, as well as interactions between channel types. N. crassa on the other hand does not maintain tip-high gradients, and clustered distributions are observed only for the stretch-activated channels. In terms of physiological roles, evidence is quite strong that the stretch-activated channels function as a growth sensor in S. ferax, but have an unknown function in N. crassa. In both organisms, the potassium permeable channels presumably function in potassium uptake. The differences between these two organisms may be due, in part, to differences in their normal environment: aquatic versus terrestrial. Copyright 1998 Academic Press. PMID- 9742194 TI - Endocytosis and membrane turnover in the germ tube of uromyces fabae AB - We have used the fluorescent dye FM4-64 as a tracer to demonstrate bulk membrane internalization (endocytosis) and redistribution of the dye within the cytoplasm of the germ tube of the rust fungus Uromyces fabae. Staining of the hyphal membrane was detected 4 s after application of FM4-64 and reached a maximum after 1 min. The highest fluorescence intensity occurred in the apex. Subsequently, staining of the plasma membrane decreased and a subapical region of the fungal protoplast (5-20 &mgr;m from the tip) displayed increasing fluorescence with a maximum after 5 min. Fluorescence in the subapical region was redistributed to an area in the hyphal tip, which corresponds to the accumulation of apical vesicles, after 10-15 min and subsequently to a cytoplasmic region in front of the two nuclei (35-45 &mgr;m from the tip). We conclude from our measurements of membrane fluorescence that the turnover time from endocytosis to secretion of the dye amounts to 15 min. The uptake of the dye into the cytoplasm, but not membrane loading, could be inhibited completely with 5 mM NaN3 or by a temperature shift to 4 degreesC. This is the first evidence for endocytosis in a fungal germ tube. Copyright 1998 Academic Press. PMID- 9742195 TI - Structure, function, and motility of vacuoles in filamentous fungi AB - Current information on the structure and function of motile tubular vacuoles in Pisolithus tinctorius and other fungi is reviewed. The use of fluorochromes to label the vacuole lumen is evaluated and observations on the structure and motility of vacuoles in P. tinctorius are differentiated from possible artifacts. The styryl dyes FM4-64 and MDY-64, used in yeast to demonstrate endocytosis, show little or no labeling of internal membranes in undamaged P. tinctorius cells. This agrees with our data showing that other probes for endocytosis such as Lucifer yellow CH are not taken up by hyphal tip cells. Overall, the observations do not support endocytosis in hyphal tips. It has been suggested that tubular vacuole systems carry out longitudinal transport, and evidence in favor of this hypothesis is evaluated. New data are presented to show that many of the large vacuoles in subapical cells are attached to the plasma membrane and are relatively immobile, while video sequences show movement of fluorochrome in pulses along a series of several large vacuoles, all interconnected via tubules. Tubular vacuoles from thick sections of hyphae processed under anhydrous conditions are shown by X-ray microanalysis to contain relatively high levels of P and K, as seen previously in the larger vacuoles. These results provide further evidence for a role of the tubular vacuoles in longitudinal transport of P. Copyright 1998 Academic Press. PMID- 9742196 TI - What determines growth direction in fungal hyphae? AB - We used high-resolution video microscopy and image analysis to map the trajectory of the Spitzenkorper in growing hyphae of Neurospora crassa and to correlate it with growth directionality. The Spitzenkorper followed a tortuous trajectory produced by a dominant forward motion accompanied by frequent, transverse oscillations. In hyphae with a fixed growth direction, the regression line of the Spitzenkorper trajectory corresponded to the longitudinal axis of the hypha. A permanent change in growth direction, i.e., the establishment of a new growth axis, was correlated with a sustained shift in Spitzenkorper trajectory away from the existing cell axis. In meandering hyphae, changes in growth directionality occurred somewhat erratically but there was a strong compensatory tendency reversing directional shifts and maintaining an overall fixed direction of growth. Although external factors greatly affect hyphal growth direction (tropisms), they are probably not the primary determinants of growth directionality. Inhibitors of microtubules, but not of actin microfilaments, caused hyphae to lose their growth directionality-providing support for the idea that Spitzenkorper trajectory is determined internally by a growing scaffolding of cytoplasmic microtubules. The meandering morphology of N. crassa hyphae was duplicated by computer simulation in support of the idea that hyphal morphogenesis is controlled by the position of the Spitzenkorper functioning as a vesicle supply center. PMID- 9742197 TI - The yeast cytoskeleton: the closer We look, the more We See AB - May, K. M., and Hyams, J. S. 1998. The yeast cytoskeleton: The closer we look, the more we see. Copyright 1998 Academic Press. PMID- 9742198 TI - The actomyosin cytoskeleton of amoebae of the cellular slime molds acrasis rosea and protostelium mycophaga: structure, biochemical properties, and function AB - In amoebae of the cellular slime molds (mycetozoans) Acrasis rosea and Protostelium mycophaga, bundles of F-actin radiate from the endoplasm-ectoplasm interface into the pseudopodia, where G-actin is also located. We conclude that these actin bundles form a core scaffold driving pseudopod extension which is subsequently completed by filling with a more loosely organized meshwork of F actin. Some bipolar, elongate amoebae of A. rosea also contained long bundles of F-actin that traverse the cells lengthwise and remotely resemble stress fibers. Rodlets of F-actin were scattered in the body of amoebae of A. rosea or formed star-shaped or polygonal complexes near or around contractile vacuoles, where they may play a role in contraction. In total protein extracts analyzed by SDS PAGE and immunoblots the actins migrated like the rabbit skeletal muscle control. The relative proportion of actin in total protein extracts was 7.9% for A. rosea and 34.5% for P. mycophaga. We detected four or five isoactins in extracts of both species and we determined that the genome of each species contains approximately six actin genes. Whether they are all expressed or if posttranslational modifications occur remains to be determined. Myosin II was enriched in actomyosin extracts; its Mr was 187.8 kDa for A. rosea and 220.7 kDa for P. mycophaga. Cell models ("ghosts") contracted upon the addition of ATP. We conclude that amoebae of A. rosea and P. mycophaga, although behaving differently from those of Dictyostelium discoideum, contain the basic repertoire of molecules that enable pseudopod extension by actin polymerization and ATP-induced contraction of the cell cortex. Copyright 1998 Academic Press. PMID- 9742200 TI - Organelle transport and molecular motors in fungi AB - Polarized growth, secretion of exoenzymes, organelle inheritance, and organelle positioning require vectorial transport along cytoskeletal elements. The discovery of molecular motors and intensive studies on their biological function during the past 3 years confirmed a central role of these mechanoenzymes in morphogenesis and development of yeasts and filamentous fungi. Saccharomyces cerevisiae proved to be an excellent model system, in which the complete set of molecular motors is presumed to be known. Genetic studies combined with cell biological methods revealed unexpected functional relationships between these motors and has greatly improved our understanding of nuclear migration, exocytosis, and endocytosis in yeasts. Tip growth of elongated hyphae, compared to budding, however, does require vectorial transport over long distances. The identification of ubiquitous motors that are not present in yeast indicates that studies on filamentous fungi might be helpful to elucidate the role of motors in long-distance organelle transport within higher eukaryotic cells. Copyright 1998 Academic Press. PMID- 9742199 TI - Mitosis in wild-type and beta-tubulin mutant strains of Aspergillus nidulans. AB - We review and illustrate the wild-type mitotic cycle of Aspergillus nidulans and report the sequence alterations in six mutant alleles of the A. nidulans benA, beta-tubulin, gene. These alleles confer heat sensitivity and resistance to the antifungal, antimicrotubule compound benomyl, and they have been very important in the study of mitosis and microtubule function in A. nidulans. The mutations are novel and fall at amino acids 50, 134, and 257. We have examined the phenotypes conferred by the mutations at restrictive temperatures. None blocks the assembly of microtubules. One allele, benA33, blocks anaphase A and partially inhibits the disassembly of cytoplasmic microtubules in mitosis. We also often observe abnormal spindle morphologies in strains carrying benA33. Another allele, benA31, causes arrest in mitosis with short mitotic spindles and, thus, appears to inhibit spindle elongation. PMID- 9742202 TI - The relationship between B-mating-type genes and nuclear migration in schizophyllum commune AB - Raudaskoski, M. 1998. The relationship between B-mating-type genes and nuclear migration in Schizophyllum commune. Copyright 1998 Academic Press. PMID- 9742203 TI - Acropetal: a genetic locus required for conidiophore architecture and pathogenicity in the rice blast fungus. AB - Fungal spores are a primary means of dissemination and are the major sources of inoculum in pathogenic species. Sporulation in the rice blast fungus Magnaporthe grisea involves the production of three-celled conidia, borne sympodially on an aerial conidiophore. A disease cycle initiates when spores are dispersed and attach to the rice plant surface. Using insertional mutagenesis we have identified a major regulator of conidiophore morphogenesis in M. grisea. A null mutation in the acropetal (ACR1) locus causes a hypermorphic conidiation phenotype where indeterminate growth of the conidial tip cell results in the production of head-to-tail (acropetal) arrays of spores. acropetal mutants are nonpathogenic and fail to undergo infection-related morphogenesis. The ACR1 locus encodes a spore-specific transcript and acr1(-) mutants fail to turn off the expression of the hydrophobin encoding gene MPG1 in dormant spores. We propose that ACR1 is a stage-specific negative regulator of conidiation that is required to establish a sympodial pattern of spore formation. Interestingly a failure to establish the correct pattern of sporulation in M. grisea results in the production of spores that cannot progress through the disease cycle. Studies of Acropetal suggest that the diverse patterns of spore ontogeny in conidial fungi arose through alterations in major genes controlling spore-specific gene expression. PMID- 9742201 TI - Dynamics of cell wall formation in fission yeast, Schizosaccharomyces pombe. AB - Studies on the dynamics of surface and intracellular structures during cell wall formation from the reverting protoplast of Schizosaccharomyces pombe were reviewed, and the correlation between cell wall formation and actin cytoskeleton, which is the most important conductor of the mechanism, is described in this paper. A close spatial and temporal relationship between actin cytoskeleton and cell wall formation was found by using wild type and actin point-mutant cps8 of S. pombe. Concomitant with the cell wall formation, dynamic behavior of the intracellular secretion machinery, especially the Golgi apparatus and secretory vesicles, was analyzed by three-dimensional reconstruction of 40 to 80 serial sections at five reverting stages. Total reverting protoplast volume increased by 3.8 and 4.3 times at 3 and 5 h, respectively, and the volume of the Golgi apparatus in the corresponding stages increased 2.3- and 2. 5-fold over the same periods. The number of secretory vesicles also markedly increased by 3.4 and 5.8 times over that of the corresponding reverting protoplasts. Actin point-mutant cps8 cells have abnormal structure in the cell wall and septum, and the distribution pattern of the actin cytoskeleton during the reversion process was different from wild-type protoplasts. The profiles of actin showed one or two thick cables and patches in the cytoplasm which remained throughout reversion. The development of crosslinkage of the glucan fibrils which are beta-1,3-glucan in nature on the reverting protoplast surface was defective; the glucan networks consisted of thin, rope-shaped fibrils up to 30 nm in width which formed a ribbon shape 200 nm wide in wild-type reverting protoplasts. The intrafibrillar space is not filled with amorphous particles of alpha-galactomannan in nature. The secretion machinery was seen to have a similar profile as the wild type. The above results suggest that actin cytoskeleton may control secretion of beta-1,6 glucan and other cell wall substances such as alpha-glucan and alpha galactomannan rather than beta-1,3-glucan. Study of the role of actin cytoskeleton in the cell wall formation is contributing to the development of antifungal agents together with basic cell biology. PMID- 9742204 TI - Melanin synthesis is associated with changes in hyphopodial turgor, permeability, and wall rigidity in gaeumannomyces graminis var. graminis. AB - Mycelia of Gaeumannomyces graminis var. graminis form large cells called hyphopodia with deeply lobed, melanized walls. Like appressoria produced by other pathogens, hyphopodia develop on hydrophobic surfaces, but it is not clear that hyphopodia function as platforms for host penetration. In appressoria, melanin synthesis is linked to the generation of enormous turgor pressures that provide the necessary force for plant penetration. In the present study, hyphopodial turgor was measured in a wild-type strain of G. graminis var. graminis, a mutant exhibiting constitutive synthesis of melanin (referred to as the dark mutant), and a melanin-deficient strain (thr). These experiments demonstrate that hyphopodia of the wild-type strain generate higher pressures than the dark mutant and that nonmelanized thr hyphopodia generate minuscule internal pressures. Melanization of the wall is also associated with an increase in its rigidity. These data correlate with differences in wall permeability consistent with a recent model for turgor generation by appressoria. PMID- 9742205 TI - Use of molecular cytology to study the structure and biology of phytopathogenic and mycorrhizal fungi. AB - Molecular cytology, that is, the in situ localization of selected molecules by labeling with lectins, enzymes, and antibodies, has made a major contribution to our understanding of the structure and biology of fungi and is increasingly becoming an integral part of molecular, genetic, and biochemical studies. The review presented in this article concentrates on recent advances in the application of molecular cytology in investigations of the structure and biology of phytopathogenic and mycorrhizal fungi and of the molecular basis of their infection of host plants. The review examines details of the structure and molecular composition of fungal cell walls revealed by lectin, enzyme, and antibody labeling. Molecular composition is shown to vary according to taxonomic relationships and as a reflection of differences in cell type, location within the cell, and within thickness of the wall. Sites of synthesis and secretion of wall components are also detected through the labeling of selected molecules. In situ labeling of cytoskeletal elements, microtubules and actin microfilaments, has provided much information on the role of these elements in tip growth, organelle distribution, and spore development. Molecular cytology, particularly through the generation of monoclonal antibodies, has also revealed new and exciting information on specialized infection structures formed by fungi in order to infect host plants. The sites of storage and secretion of adhesives and degradative enzymes have been documented, as have surface specializations that may be associated with avoidance of detection by the host. In addition, in situ labeling with enzymes and antibodies has aided studies of the host defense response, including mechanisms of detection of fungal elicitor molecules, changes in wall composition, and the secretion of antifungal compounds. With the increasing production of monoclonal antibodies to fungal molecules, molecular cytology promises to continue to make an important contribution to our understanding of fungal cell structure and function in the future. PMID- 9742207 TI - Signalling through the leukotriene B4 receptor involves both alphai and alpha16, but not alphaq or alpha11 G-protein subunits. AB - COS-7 cells transfected with the leukotriene (LT) B4 receptor (BLTR) cDNA were unable to produce LTB4-induced inositol phosphates (IPs) in spite of the presence of endogenous Galphai, Galphaq and Galpha11 proteins. Co-transfection of BLTR with Galpha16, however, resulted in high levels of IP production, which were 17-, 10- and 6-fold higher than with co-transfected Galpha11, Galphaq and Galpha14, respectively. Co-transfection of BLTR with phospholipase C (PLC) beta2, on the other hand, resulted in efficient IP production and co-transfection of BLTR with both Galpha16 and PLCbeta2 resulted in a greater than additive response. PMID- 9742208 TI - Insulin-stimulated expression of c-fos, fra1 and c-jun accompanies the activation of the activator protein-1 (AP-1) transcriptional complex. AB - The activator protein-1 (AP-1) transcriptional complex is made up of members of the Fos (c-Fos, FosB, Fra1, Fra2) and Jun (c-Jun, JunB, JunD) families and is stimulated by insulin in several cell types. The mechanism by which insulin activates this complex is not well understood but it is dependent on the activation of the Erk1 and Erk2 isoforms of mitogen-activated protein kinases. In the current study we show that the AP-1 complex isolated from insulin-stimulated cells contained c-Fos, Fra1, c-Jun and JunB. The activation of the AP-1 complex by insulin was accompanied by (i) a transient increase in c-fos expression, and the transactivation of the ternary complex factors Elk1 and Sap1a, in an Erk1/Erk2-dependent fashion; (ii) a substantial increase in the expression of Fra1 protein and mRNA, which was preceded by a transient decrease in its electrophoretic mobility upon SDS/PAGE, indicative of phosphorylation; and (iii) a sustained increase in c-jun expression without increasing c-Jun phosphorylation on serines 63 and 73 or activation of the stress-activated kinase JNK/SAPK. In conclusion, insulin appears to stimulate the activity of the AP-1 complex primarily through a change in the abundance of the components of this complex, although there may be an additional role for Fra1 phosphorylation. PMID- 9742206 TI - Protein kinase B (c-Akt): a multifunctional mediator of phosphatidylinositol 3 kinase activation. AB - While a plethora of extracellular molecules exist that modulate cellular functions via binding to membrane receptors inside the cell, their actions are mediated by relatively few signalling mechanisms. One of these is activation of phosphatidylinositol 3-kinase (PI-3K), which results in the generation of a membrane-restricted second messenger, polyphosphatidylinositides containing a 3' phosphate. How these molecules transduced the effects of agonists of PI-3K was unclear until the recent discovery that several protein kinases become activated upon exposure to 3'-phosphorylated inositol lipids. These enzymes include protein kinase B (PKB)/AKT and PtdIns(3,4, 5)P3-dependent kinases 1 and 2, the first two of which interact with 3'-phosphorylated phosphoinositides via pleckstrin homology domains. Once targeted to the membrane by this motif, PKB becomes phosphorylated at two residues, which relieves intermolecular inhibition, allowing the activated complex to dissociate and modify its targets. Identification of these substrates is the subject of intensive research, since at least one must play a key role in suppressing apoptosis, as demonstrated by expression of activated alleles of PKB. The generation of effective transdominant mutants, coupled with genetic analysis of the protein kinase in simpler organisms, should help in elucidating outstanding questions in the functions, targets and regulation of this important mediator of PI-3K signalling. PMID- 9742209 TI - Sulphoxidation reaction catalysed by myeloperoxidase from human leucocytes. AB - The oxidation of alkyl aryl sulphides by myeloperoxidase (MPO) at the expense of hydrogen peroxide was investigated under steady-state conditions. The sulphide concentration effect was studied under saturating H2O2 concentrations at pH 5.0 and 20 degreesC. The kinetic constants, kcat and Km, of the different substrates were determined and the values were in the 1-10 s-1 range and around 43+/-26 microM respectively, whatever the sulphide considered. In the case of p substituted thioanisoles, the oxidation rate was dependent upon the substituent effect. The correlation of log(kcat) with the substituent constants (sigma+ values) (Hammett equation) could be explained by a reaction mechanism involving the enzyme compound II and a sulphenium radical cation. This conclusion was also supported by spectrophotometric analysis of catalytic intermediates of the enzyme, showing the accumulation of compound II. Moreover, chiral HPLC analyses showed that MPO oxidation of alkyl aryl sulphides produced the corresponding (R) sulphoxides with a low enantioselectivity (4-8%). Chloride ion effects on the MPO catalysed oxygenation of sulphides were also studied. Chloride acted as a substrate for MPO and as an activator in MPO-catalysed sulphoxidation. Inhibition occurred at chloride concentrations above 120 mM, whereas below 120 mM, chloride increased the reaction rate when using p-tolyl methyl sulphide as the substrate. In the presence of 100 mM chloride the catalytic efficiency (kcat/Km) of MPO increased 3-4-fold, whatever the sulphide considered, but racemic products were obtained. These data have been interpreted in the light of known structural information on the accessibility of the distal haem cavity. PMID- 9742210 TI - Localization of a cyclopentenone prostaglandin to the endoplasmic reticulum and induction of BiP mRNA. AB - Cyclopentenone prostaglandins (PGs) are transported into cells and stimulate the expression of various stress genes, such as that coding for BiP (an ER luminal protein). To reveal the site of action of the PGs for the induction of stress gene expression, we introduced a fluorescent probe, pyrene, into two types of PG analogue, GIF0010 (a cyclopentenone type) and GIF0037 (a cyclopentanone type) and examined their intracellular localization in normal rat kidney cells and their ability to induce the BiP gene expression. GIF0010 accumulated around the nuclei and coincided with BiP, a resident protein in the endoplasmic reticulum (ER) and markedly induced BiP gene expression. By contrast, GIF0037 and pyrene neither accumulated in the cell nor induced BiP gene expression. Thus the ER localization of GIF0010 and the induction of gene expression by GIF0010 are ascribed to the cyclopentenone structure. Treatment with cycloheximide inhibited both the accumulation of GIF0010 and the induction of the BiP mRNA, suggesting that the ER localization of the PG and subsequent gene expression require the nascent protein synthesis. These results demonstrate that the cyclopentenone PG is specifically accumulated in the ER, transducing a signal for BiP gene expression in the nuclei. PMID- 9742211 TI - Characterization of p96h2bk: immunoreaction with an anti-Erk(extracellular-signal regulated kinase) peptide antibody and activity in Xenopus oocytes and eggs. AB - We have shown previously that oncogenic Ras induces cell cycle arrest in activated Xenopus egg extracts [Pan, Chen and Lin (1994) J. Biol. Chem. 269, 5968 5975]. The cell cycle arrest correlates with the stimulation of a protein kinase activity that phosphorylates histone H2b in vitro (designated p96(h2bk)) [Chen and Pan (1994) J. Biol. Chem. 269, 28034-28043]. We report here that p96(h2bk) is likely to be p96(ram), a protein of approx. 96 kDa that immunoreacts with a monoclonal antibody (Mk-1) raised against a synthetic peptide derived from a sequence highly conserved in Erk1/Erk2 (where Erk is extracellular-signal regulated kinase). This is supported by two lines of evidence. First, activation/inactivation of p96(h2bk) correlates with upward/downward bandshifts of p96(ram) in polyacrylamide gels. Secondly, both p96(h2bk) and p96(ram) can be immunoprecipitated by antibody Mk-1. We also studied the activity of p96(h2bk)/p96(ram) in Xenopus oocytes and eggs. p96(h2bk)/p96(ram) was inactive in stage 6 oocytes, was active in unfertilized eggs, and became inactive again in eggs after fertilization. Since stage 6 oocytes are at G2-phase of the cell cycle, unfertilized eggs arrest at M-phase and eggs exit M-phase arrest after fertilization, the results thus indicate that p96(h2bk)/p96(ram) activity is cell cycle dependent. Moreover, microinjection of oncogenic Ras into fertilized eggs at the one-cell stage arrests the embryos at the two-cell stage, and this induced arrest is correlated with an inappropriate activation of p96(h2bk)/p96(ram). The data are consistent with the concept that inappropriate activation of p96(h2bk)/p96(ram) plays a role in the cell cycle arrest induced by oncogenic Ras. PMID- 9742213 TI - Chondrocyte-mediated catabolism of aggrecan: aggrecanase-dependent cleavage induced by interleukin-1 or retinoic acid can be inhibited by glucosamine. AB - A rat chondrosarcoma cell line and bovine cartilage explants have been used to study the control of aggrecan degradation by chondrocytes treated with interleukin-1 (IL-1) or retinoic acid (RA). Aggrecan fragment analysis with anti neo-epitope antibodies suggests that aggrecanase (an as yet unidentified enzyme) is the only aggrecan-degrading proteinase active in these cultures. With rat cells, aggrecanase converts the aggrecan core protein into two major G1-domain bearing products (60 kDa with a C-terminal Glu-373, and 220 kDa with a C-terminal Glu-1459). Both products were quantified on a standardized Western analysis system with a G1-specific antibody. Immunoblots were analysed by scanning densitometry and the sensitivity, linearity and reproducibility of the assay were established. With rat cells the aggrecanase response to IL-1 was optimal at about 2 mM glutamine, but was progressively inhibited at higher concentrations, with about 90% inhibition at 10 mM glutamine. Such inhibition by glutamine was not, however, observed with bovine explants. On the other hand, marked inhibition of aggrecanase-dependent cleavage was observed with both rat cells and bovine explants when d(+)-glucosamine was included at concentrations above 2 mM. Inhibition was apparently not due to cytotoxicity or interference with IL-1 signalling, since biosynthetic activity was not inhibited and inhibition of the aggrecanase response was also obtained when RA was used as the catabolic stimulator. Possible mechanisms for the inhibition of the aggrecanase response by glucosamine in chondrocytes treated with IL-1 or RA are discussed. PMID- 9742212 TI - Human soluble guanylate cyclase: functional expression and revised isoenzyme family. AB - Soluble guanylate cyclase (sGC), a heterodimeric (alpha/beta) haem protein that converts GTP to the second messenger cGMP, functions as the receptor for nitric oxide (NO) and nitrovasodilator drugs. Three distinct cDNA species of each subunit (alpha1-alpha3, beta1-beta3) have been reported from various species. From human sources, none of these have been expressed as functionally active enzyme. Here we describe the expression of human alpha/beta heterodimeric sGC in Sf9 cells yielding active recombinant enzyme that was stimulated by the nitrovasodilator sodium nitroprusside or the NO-independent activator 3-(5' hydroxymethyl-2'-furyl)-1-benzylindazole (YC-1). At the protein level, both alpha and beta subunits were detected in human tissues, suggesting co-expression also in vivo. Moreover, resequencing of the human cDNA clones [originally termed alpha3 and beta3; Giuili, Scholl, Bulle and Guellaen (1992) FEBS Lett. 304, 83 88] revealed several sequencing errors in human alpha3; correction of these eliminated major regions of divergence from rat and bovine alpha1. As human beta3 also displays more than 98% similarity to rat and bovine beta1 at the amino acid level, alpha3 and beta3 represent the human homologues of rat and bovine alpha1 and beta1, and the isoenzyme family is decreased to two isoforms for each subunit (alpha1, alpha2; beta1, beta2). Having access to the human key enzyme of NO signalling will now permit the study of novel sGC-modulating compounds with therapeutic potential. PMID- 9742214 TI - Synthesis in Escherichia coli of two smaller enzymically active analogues of Coxiella burnetii macrophage infectivity potentiator (CbMip) protein utilizing a single open reading frame from the cbmip gene. AB - FK506-binding proteins (FKBPs) have been identified in a variety of eukaryotic and prokaryotic organisms. Macrophage infectivity potentiator (CbMip, 23.5 kDa) protein of the obligate intracellular bacterium, Coxiella burnetii, was shown previously to belong to the family of FKBPs based on sequence homology and peptidyl-prolyl cis/trans isomerase (PPIase) activity. Further characterization of the cbmip gene has identified two additional proteins with molecular masses of 15.5 and 15.0 kDa that are synthesized, in addition to the 23.5 kDa CbMip, when expressed in Escherichia coli. Amino acid sequencing at the N-terminus combined with transcription and translation fusion expression revealed that the two proteins were synthesized from the same open reading frame of the cbmip gene, but starting at different internal translation start codons, probably by translational reinitiation. When the internal methionines serving as start sites were replaced with lysine by site-directed mutagenesis, the synthesis of 15.5 and 15.0 kDa proteins was abolished even though the synthesis of 23.5 kDa CbMip was intact. This confirmed that the 15.5 and 15.0 kDa proteins are indeed generated by translational reinitiation and are not degradation products of the 23.5 kDa protein. Like other FKBPs, both 15.5 and 15.0 kDa proteins exhibit PPIase activity. Because they share significant sequence homology with FKBPs and have a similar PPIase activity, 15.5 and 15. 0 kDa proteins are designated as C. burnetii FKBP (Cb-FKBP) analogues I and II, respectively. TnphoA mutagenesis demonstrated that whereas the large protein (CbMip) is secreted, Cb-FKBP analogues I and II are cytoplasmic, indicating that structural variations could allow for different subcellular compartmentalization of similar proteins. Western blot analysis of lysates of purified C. burnetii using a CbMip-specific monoclonal antibody revealed the presence of a protein migrating at approximately 15 kDa, indicating the presence of smaller Cb-FKBP analogue(s) in C. burnetii, although at much lower levels compared with 23.5 kDa CbMip. This unique gene organization seen with cbmip may provide the organism with a mechanism of efficient use of its limited genetic information to synthesize proteins that are structurally different yet functionally similar. PMID- 9742215 TI - Transcription of the juvenile hormone esterase gene under the control of both an initiator and AT-rich motif. AB - The binding of transcription factors to the core promoter of the juvenile hormone esterase gene was functionally characterized using both a cell-free in vitro transcription functional assay and a cell transfection assay. A core JHE promoter (-61 to +28 bp relative to transcription start site) supported faithful transcription from the in vivo transcription start site. The nuclear extracts from the Sf9 insect cell line that provided transcription from that template also bound to that template as a probe in gel-mobility shift assays. Deletion or transversion of the initiator-binding motif (-1 to +4 bp) abolished detectable transcription either in vitro or in transfected cells. An AT-rich motif (ATATAT; 28 to -23 bp) serves another transcription factor-binding site. Mutation of the AT-rich motif to a canonical TATA-box preserved transcription, while either its deletion or complete transversion abolished or significantly reduced detectable transcriptional activity. These results indicate that, under these conditions, the functional operation of this core promoter approaches that of a composite promoter in which both the TATA- and initiator-binding protein complexes are necessary, even for basal transcription. On the other hand, these debilitating mutations to either the TATA box or initiator motif did not prevent the ability of the corresponding gel-shift competitive probes to compete with the wild-type promoter for binding by the transcription factors. Even a double transversion of both the AT-rich motif and the initiator-binding motif was able to competitively displace the protein complex that bound to the labelled wild-type probe. These data strongly indicate the presence of (an) additional core-promoter-associated transcription factor(s) (that is not the 'downstream element') that contact(s) the AT-binding complex and/or initiator-binding factor with sufficient avidity to remove them from binding to the competing wild-type promoter sequence. PMID- 9742216 TI - Hydrogen peroxide-induced DNA damage is independent of nuclear calcium but dependent on redox-active ions. AB - In cells undergoing oxidative stress, DNA damage may result from attack by .OH radicals produced by the Fenton reaction, and/or by nucleases activated by nuclear calcium. In the present study, the participation of these two mechanisms was investigated in HeLa cells. Nuclear-targeted aequorin was used for selectively monitoring Ca2+ concentrations within the nuclei ([Ca2+]n), in conjunction with the cell-permeant calcium chelator bis-(o-aminophenoxy)ethane N,N,N', N'-tetraacetic acid acetoxymethyl ester (BAPTA/AM), the lipid-soluble broad-spectrum metal chelator with low affinity for Ca2+ and Mg2+ N,N,N',N' tetrakis(2-pyridylmethyl)ethylenediamine (TPEN), and the high-affinity iron/copper chelator 1, 10-phenanthroline (PHE). In Ca2+-containing medium, H2O2 induced extensive DNA strand breaks and an increase in [Ca2+]n that was almost identical to that observed in the cytosol ([Ca2+]c). In cells bathed in Ca2+ free/EGTA medium, in which the increases in [Ca2+]n and [Ca2+]c due to H2O2 were significantly reduced, similar levels of DNA fragmentation also occurred. In cells preloaded with BAPTA/AM or TPEN, the small increase of [Ca2+]n normally elicited by H2O2 in Ca2+-free medium was completely buffered, and DNA damage was largely prevented. On the other hand, pretreatment with PHE did not affect the calcium response in the nuclei, but completely prevented DNA strand breakage induced by H2O2. Re-addition of 100 microM CuSO4 and 100 microM FeSO4 to TPEN- and PHE-treated cells prior to H2O2 challenge reversed the effect of TPEN and PHE, whereas 1 mM was necessary to negate the effect of BAPTA/AM. The levels of DNA strand breakage observed, however, did not correlate with the amounts of 8 hydroxy 2'-deoxyguanosine (8-OHdG): H2O2 did not produce 8-OHdG, whereas PHE alone slightly increased 8-OHdG levels. CuSO4 and FeSO4 enhanced the effects of PHE, particularly in the presence of H2O2. Exposure of cells to a mixture of CuSO4/FeSO4 also resulted in a significant increase in 8-OHdG levels, which was prevented in cells preloaded with BAPTA/AM. Similar results were obtained in a cell-free system using isolated calf thymus DNA exposed to CuSO4/FeSO4, regardless of whether H2O2 was present or not. These results suggest that BAPTA/AM prevents H2O2-induced DNA damage by acting as an iron/copper chelator. These data also indicate that caution must be exercised in using Ca2+ chelating agents as evidence for a role in cellular Ca2+ levels in experimental conditions in which transition-metal-ion-mediated oxidant production is also occurring. PMID- 9742217 TI - The 'aromatic patch' of three proximal residues in the human acetylcholinesterase active centre allows for versatile interaction modes with inhibitors. AB - The role of the functional architecture of the human acetylcholinesterase (HuAChE) active centre in accommodating the non-covalent inhibitors tacrine and huperzine A, or the carbamates pyridostigmine and physostigmine, was analysed using 16 mutants of residues lining the active-centre gorge. Despite the structural diversity of the ligands, certain common properties of the complexes could be observed: (a) replacement of aromatic residues Tyr133, Tyr337 and especially Trp86, resulted in pronounced changes in stability of all the complexes examined; (b) effects due to replacements of the five other aromatic residues along the active-centre gorge, such as the acyl pocket (Phe295, Phe297) or at the peripheral anionic site (Tyr124, Trp286, Tyr341) were relatively small; (c) effects due to substitution of the carboxylic residues in the gorge (Glu202, Glu450) were moderate. These results and molecular modelling indicate that the aromatic side chains of residues Trp86, Tyr133 and Tyr337 form together a continuous 'aromatic patch' lining the wall of the active-centre gorge, allowing for the accommodation of the different ligands via multiple modes of interaction. Studies with HuAChE mutants carrying replacements at positions 86, 133 and 337 indicate that the orientations of huperzine A and tacrine in the HuAChE complexes in solution are significantly different from those observed in X-ray structures of the corresponding complexes with Torpedo californica AChE (TcAChE). These discrepancies may be explained in terms of structural differences between the complexes of HuAChE and TcAChE or, more likely, by the enhanced flexibility of the AChE active-centre gorge in solution as compared with the crystalline state. PMID- 9742218 TI - SH2-Balpha is an insulin-receptor adapter protein and substrate that interacts with the activation loop of the insulin-receptor kinase. AB - We identified SH2-Balpha as an insulin-receptor-binding protein based on interaction screening in yeast hybrid systems and co-precipitation in cells. SH2 Balpha contains pleckstrin-homology ('PH') and Src homology 2 (SH2) domains and is closely related to APS (adapter protein with a PH domain and an SH2 domain) and lnk, adapter proteins first identified in lymphocytes. SH2-Balpha is ubiquitously expressed and is present in rat epididymal adipose tissue, liver and skeletal muscle, physiological sites of insulin action. On SDS/PAGE, SH2-Balpha migrates at a molecular mass of 98 kDa, although the predicted size of SH2-Balpha is 79.6 kDa. Insulin causes an electrophoretic mobility shift. SH2-Balpha can be immunoprecipitated using anti-(insulin receptor) antibody from insulin-stimulated cells. Anti-phosphotyrosine antibody or the growth factor receptor-binding protein 2 (Grb2) SH2 domain precipitate SH2-Balpha after insulin stimulation, suggesting that SH2-Balpha is tyrosine-phosphorylated and may be a substrate for the insulin receptor. The SH2-Balpha SH2 domain did not interact with insulin receptor substrate (IRS) proteins or epidermal-growth-factor receptor. Mutation of the juxtamembrane and C-terminus of the insulin receptor did not abolish the interaction with the SH2 domain. This was further confirmed using a panel of activation-loop single point mutants where mutation of Tyr1158, Tyr1162 and Tyr1163 abolished interaction. Thus SH2-Balpha is a likely component in the insulin-signalling pathway and may function as an alternative signalling protein by interacting with the activation loop of the insulin-receptor cytoplasmic domain. PMID- 9742219 TI - Cloning and expression of mouse legumain, a lysosomal endopeptidase. AB - Legumain, a recently discovered mammalian cysteine endopeptidase, was found in all mouse tissues examined, but was particularly abundant in kidney and placenta. The distribution in subcellular fractions of mouse and rat kidney showed a lysosomal localization, and activity was detectable only after the organelles were disrupted. Nevertheless, ratios of legumain activity to that of cathepsin B differed considerably between mouse tissues. cDNA encoding mouse legumain was cloned and sequenced, the deduced amino acid sequence proving to be 83% identical to that of the human protein [Chen, Dando, Rawlings, Brown, Young, Stevens, Hewitt, Watts and Barrett (1997) J. Biol. Chem. 272, 8090-8098]. Recombinant mouse legumain was expressed in human embryonic kidney 293 cells by use of a vector containing a cytomegalovirus promoter. The recombinant enzyme was partially purified and found to be an asparagine-specific endopeptidase closely similar to naturally occurring pig kidney legumain. PMID- 9742220 TI - Mixed-lineage kinase 2-SH3 domain binds dynamin and greatly enhances activation of GTPase by phospholipid. AB - Mixed-lineage kinase 2 (MLK2) is a cytoplasmic protein kinase expressed at high levels in mammalian brain. The MLK2 structure is composed of a Src homology 3 (SH3) domain, two leucine zippers, a basic motif, a Cdc42/Rac interactive binding motif and a large C-terminal domain rich in proline, serine and threonine residues. To begin to define the role of MLK2 in mammalian brain, we used an MLK2 SH3 domain-glutathione S-transferase fusion protein (GST-MLK2-SH3) to isolate MLK2-binding proteins from rat brain extract. This analysis revealed that the major MLK2-SH3-domain-binding protein in rat brain is the GTPase dynamin. By using two different forms of the dynamin proline-rich domain as affinity ligands, the binding site for MLK2-SH3 was mapped to the C-terminal region of dynamin between residues 832 and 864. In GTPase assays, the addition of MLK2-SH3 stimulated the activity of purified dynamin I by 3-fold over the basal level, whereas the addition of a known dynamin activator, phosphatidylserine (PtdSer), stimulated a 6-fold increase. When MLK2-SH3 was added to the assay together with PtdSer, however, dynamin GTPase activity accelerated by more than 23-fold over basal level. An MLK2 mutant (MLK2-W59A-SH3), with alanine replacing a conserved tryptophan residue in the SH3 domain consensus motif, had no effect on dynamin activity, either alone or in the presence of PtdSer. In the same assay the SH3 domain from the regulatory subunit of phosphatidylinositol 3'-kinase stimulated a similar synergistic acceleration of dynamin GTPase activity in the presence of PtdSer. These results suggest that synergy between phospholipid and SH3 domain binding might be a general mechanism for the regulation of GTP hydrolysis by dynamin. PMID- 9742221 TI - Functional properties of a naturally occurring isoform of soluble guanylyl cyclase. AB - Soluble guanylyl cyclase (sGC), the target enzyme of the signalling molecule NO, contains one prosthetic haem group and consists of an alpha and a beta subunit. So far, only the alpha1beta1 heterodimer has been shown to exist in different cells and tissues, and most biochemical studies of sGC have been performed with the alpha1 beta1 heterodimer. Here we demonstrate for the first time the natural occurrence of the alpha2 subunit on the protein level. The alpha2 subunit co precipitated with the beta1 subunit from human placenta, showing the existence of the alpha2 beta1 isoform in vivo. The new enzyme was expressed in and purified from cells from the Spodoptera frugiperda ovary cell line Sf 9. Spectral analysis showed that the alpha2 beta1 heterodimer contains a prosthetic haem group revealing the same characteristics as the haem in the alpha1 beta1 form. The kinetic properties of both isoforms and sensitivity towards NO were indistinguishable. 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), a selective inhibitor of sGC, abolished NO-stimulated activity of both heterodimers. The new NO-independent activator, 3-(5'-hydroxymethyl-2'-furyl)-1-benzyl indazole (YC-1), increased the maximal NO-stimulated activity of the new isoform, caused a leftward-shift in the NO concentration-response curve and turned CO into an effective activator, as it did for the alpha1 beta1 heterodimer (200-fold activation). In summary, the differences in primary structure of both alpha subunits are contrasted by their functional similarity. Further studies will be needed to elucidate the physiological purpose of the new isoform. PMID- 9742222 TI - Mutational analysis of trans-membrane helices M3, M4, M5 and M7 of the fast twitch Ca2+-ATPase. AB - Mutational analysis of trans-membrane helices M3, M4, M5 and M7 of the Ca2+ ATPase revealed a novel phenotypic variant, M4 [Y295A (the one-letter symbols are used for amino acid residues throughout)], displaying an increased affinity for Pi and decreased affinity for MgATP, while retaining the ability to translocate Ca2+ ions across the endoplasmic reticulum membrane. The properties of this mutant suggest that the E1-E2 equilibrium is shifted towards E2, and indicate a key role for this aromatic residue (Y295) at the end of trans-membrane helix M4. A mutant containing three amino acid residue substitutions at the end of the seventh trans-membrane helix, M7 (F834A, F835A, T837F), showed a complete loss of ATPase activity and a reduced ability to phosphorylate with Pi, although MgATP initiated phosphorylation was unaffected. The observation that single mutations in this cluster of residues had no effect on Ca2+ transport suggests that correct anchoring of the helix at the lipid-water interface by these aromatic residues is important in the functioning of the ATPase. Mutation of polar residues in helix M3 did not affect inhibition of the ATPase by thapsigargin, thapsivillosin A or t butyl hydroquinone, suggesting that hydrogen-bonding partners for the essential OH groups on these inhibitors lie elsewhere in the ATPase. PMID- 9742223 TI - Nerve growth factor- and epidermal growth factor-stimulated translocation of the ADP-ribosylation factor-exchange factor GRP1 to the plasma membrane of PC12 cells requires activation of phosphatidylinositol 3-kinase and the GRP1 pleckstrin homology domain. AB - ADP-ribosylation factors (ARFs) are small GTP-binding proteins that are regulators of vesicle trafficking in eukaryotic cells. GRP1 is a member of a family of ARF guanine-nucleotide-exchange factors that binds in vitro the lipid second messenger phosphatidylinositol 3,4, 5-trisphosphate [PtdIns(3,4,5)P3]. In order to study the effects of PtdIns(3,4,5)P3 on the function of GRP1, we have cloned the human homologue of GRP1, encoding for a protein which is 98.8% identical to mouse brain GRP1. Human GRP1 binds, via its pleckstrin homology (PH) domain, the inositol head group of PtdIns(3,4,5)P3, inositol 1, 3,4,5 tetrakisphosphate [Ins(1,3,4,5)P4], with high affinity (Kd 32. 2+/-5.2 nM) and inositol phosphate specificity [Kd values for Ins(1, 3,4,5,6)P5, InsP6, Ins(1,3,4)P3 and Ins(1,4,5)P3: 283+/-32, >10000, >10000 and >10000 nM, respectively). Furthermore, GRP1 can accommodate addition of glycerol or diacetylglycerol to the 1-phosphate of Ins(1,3,4,5)P4, data that are consistent with its proposed role as a putative PtdIns(3,4,5)P3 receptor. To address whether GRP1 binds PtdIns(3,4,5)P3 in vivo, we have expressed a chimaera of green fluorescent protein (GFP) fused to the N-terminus of GRP1 in PC12 cells and, using confocal microscopy, examined its resultant localization in live cells. Stimulation with either nerve growth factor or epidermal growth factor (both at 100 ng/ml) results in a rapid, PH-domain dependent, translocation of GFP-GRP1 from the cytosol to the plasma membrane, which occurs with a time course that parallels the production of PtdIns(3,4,5)P3. This translocation is dependent on the activation of phosphatidylinositol 3-kinase, since it is inhibited by wortmannin (100 nM), LY294002 (50 microM) and by the co-expression with dominant negative p85. Taken together these data strongly suggest that GRP1 interacts in vivo with plasma membrane-located PtdIns(3,4,5)P3 and hence constitutes a true PtdIns(3,4,5)P3 receptor. PMID- 9742225 TI - Cobalamin (vitamin B12) biosynthesis: identification and characterization of a Bacillus megaterium cobI operon. AB - A 16 kb DNA fragment has been isolated from a Bacillus megaterium genomic library and fully sequenced. The fragment contains 15 open reading frames, 14 of which are thought to constitute a B. megaterium cobalamin biosynthetic (cob) operon. Within the operon, 11 genes display similarity to previously identified Salmonella typhimurium cobalamin biosynthetic genes (cbiH60, -J, -C, -D, -ET, -L, -F, -G, -A, cysGA and btuR), whereas three do not (cbiW, -X and -Y). The genes of the B. megaterium cob operon were compared with the cobalamin biosynthetic genes of Pseudomonas denitrificans, Methanococcus jannaschii and Synechocystis sp. Taking into account the presence of cbiD and cbiG, the absence of a cobF, cobG and cobN, -S and -T, it was concluded that B. megaterium, M. jannaschii and Synechocystis sp., like S. typhimurium, synthesize cobalamin by an anaerobic pathway, in which cobalt is added at an early stage and molecular oxygen is not required. PMID- 9742224 TI - Functional domains of interferon regulatory factor I (IRF-1). AB - Interferon (IFN) regulatory factors (IRFs) are a family of transcription factors among which are IRF-1, IRF-2, and IFN consensus sequence binding protein (ICSBP). These factors share sequence homology in the N-terminal DNA-binding domain. IRF-1 and IRF-2 are further related and have additional homologous sequences within their C-termini. Whereas IRF-2 and ICSBP are identified as transcriptional repressors, IRF-1 is an activator. In the present work, the identification of functional domains in murine IRF-1 with regard to DNA-binding, nuclear translocation, heterodimerization with ICSBP and transcriptional activation are demonstrated. The minimal DNA-binding domain requires the N-terminal 124 amino acids plus an arbitrary C-terminal extension. By using mutants of IRF-1 fusion proteins with green fluorescent protein and monitoring their distribution in living cells, a nuclear location signal (NLS) was identified and found to be sufficient for nuclear translocation. Heterodimerization was confirmed by a two hybrid system adapted to mammalian cells. The heterodimerization domain in IRF-1 was defined by studies in vitro and was shown to be homologous with a sequence in IRF-2, suggesting that IRF-2 also heterodimerizes with ICSBP through this sequence. An acidic domain in IRF-1 was found to be required and to be sufficient for transactivation. Epitope mapping of IRF-1 showed that regions within the NLS, the heterodimerization domain and the transcriptional activation domain are exposed for possible contacts with interacting proteins. PMID- 9742226 TI - Cobalamin (vitamin B12) biosynthesis: functional characterization of the Bacillus megaterium cbi genes required to convert uroporphyrinogen III into cobyrinic acid a,c-diamide. AB - The function of individual genes of the Bacillus megaterium cobI operon genes in cobalamin (vitamin B12) biosynthesis was investigated by their ability to complement defined Salmonella typhimurium cob mutants. This strategy confirmed the role of cbiA, -D, -F, -J, -L and cysGA. Furthermore the operon as a whole was used to restore corrin biosynthesis in Escherichia coli, which, although closely related to S. typhimurium, does not possess the CobI pathway. When the B. megaterium cob operon was cloned into a plasmid and transformed into an E. coli strain containing the S. typhimurium cbiP, it conferred upon the host strain the ability to make the cobyric acid de novo. However, cobyric acid synthesis was observed only when the strain was grown anaerobically. Derivatives of the corrin producing E. coli strain were constructed in which genes of the B. megaterium cob operon had been inactivated. These strains were used to demonstrate that, whereas B. megaterium cbiD, -G and -X are essential for cobyric acid synthesis, the cbiW and -Y genes could be deleted without detriment to cobyric acid production in E. coli. PMID- 9742227 TI - Evidence that mammalian phosphatidylinositol transfer protein regulates phosphatidylcholine metabolism. AB - Phosphatidylinositol transfer proteins (PITPs) and their yeast counterpart (SEC14p) possess the ability to bind phosphatidylinositol (PtdIns) and transfer it between membranes in vitro. However, the biochemical function of these proteins in vivo is unclear. In the present study, the physiological role of PITP was investigated by determining the biochemical consequences of lowering the cellular content of this protein. WRK-1 rat mammary tumour cells were transfected with a plasmid containing a full-length rat PITPalpha cDNA inserted in the antisense orientation and the resultant cell clones were analysed. Three clones expressing antisense mRNA for PITPalpha were compared with three clones transfected with the expression vector lacking the insert. The three antisense clones had an average of 25% less PITPalpha protein than control clones. Two of the three antisense clones also exhibited a decreased rate of growth. All three antisense clones exhibited a significant decrease in the incorporation of labelled precursors into PtdCho during a 90-min incubation period. Under the same conditions, however, there was no change in precursor incorporation into PtdIns. Further experimentation indicated that the decrease in precursor incorporation seen in antisense clones was not due to an increased rate of turnover. When choline metabolism was analysed more extensively in one control (2-5) and one antisense (4-B) clone using equilibrium-labelling conditions (48 h of incubation), the following were observed: (1) the decrease in radioactive labelling of PtdCho seen in short-term experiments was also observed in long-term experiments, suggesting that the total amount of PtdCho was lower in antisense transfected clones (this was confirmed by mass measurements); (2) a similar decrease was seen in cellular sphingomyelin, lysoPtdCho and glycerophosphorylcholine; (3) an average two-fold increase in cellular phosphorylcholine was observed in the antisense-transfected clone; (4) cellular choline was, on average, decreased; and (5) cellular CDPcholine was not significantly altered. PMID- 9742228 TI - Activation mechanism and modification kinetics of Chinese hamster dihydrofolate reductase by p-chloromercuribenzoate. AB - Substrate effects on the activation kinetics of Chinese hamster dihydrofolate reductase by p-chloromercuribenzoate (pCMB) have been studied. On the basis of the kinetic equation of substrate reaction in the presence of pCMB, all modification kinetic constants for the free enzyme and enzyme-substrate binary and ternary complexes have been determined. The results of the present study indicate that the modification of Chinese hamster dihydrofolate reductase by pCMB shows single-phase kinetics, and that changes in the enzyme activity and tertiary structure proceed simultaneously during the modification process. Both substrates, NADPH and 7,8-dihydrofolate, protect dihydrofolate reductase against modification by pCMB. In the presence of a saturating concentration of NADPH, the value of kcat for 7,8-dihydrofolate in the enzyme-catalysed reaction increased four-fold on modification of Cys-6, accompanied by a two-fold increase in Km for the modified enzyme. The utilization of the binding energy of a group to increase kcat rather than reduce Km implies that the full binding energy of the group is not realized in the formation of the enzyme-substrate complex, but is used to stabilize the enzyme-transition-state complex. PMID- 9742230 TI - Synthesis and pairing properties of oligoribonucleotide analogues containing a metal-binding site attached to beta-D-allofuranosyl cytosine. AB - A method for the facile preparation of oligoribonucleotide analogues containing beta-d-allofuranosyl nucleosides with additional functional groups tethered to the 6'-O positions is presented. It is based on the synthesis of two protected nucleosides carrying a 6'-O -bromopentyl and a 6'-O -methylaminopentyl substituent. By a simple two-step procedure, these key intermediates were transformed into two phosphoramidites carrying a 1-aza-18-crown-6 and a triethyleneglycol group, respectively, each capable of complexing metal ions. By automated synthesis, these functionalized nucleoside analogues were efficiently incorporated into short oligoribonucleotides. Under physiological conditions (150 mM NaCl, 2 mM MgCl2, pH 7.4), incorporation of a single allofuranosyl cytosine substituted with a triethyleneglycol moiety led to a significant enthalpic stabilization of an A-type RNA duplex. This observation is in agreement with a metal ion-mediated stabilizing interaction between the two pairing strands. PMID- 9742229 TI - The use of diaminopurine to investigate structural properties of nucleic acids and molecular recognition between ligands and DNA. AB - 2,6-Diaminopurine (DAP) is an analogue of adenine which can be converted to nucleotides that serve as substrates for incorporation into nucleic acids by polymerases in place of (d)AMP. It pairs with thymidine (or uracil), engaging in three hydrogen bonds of the Watson-Crick type. The result of DAP incorporation is to add considerable stability to the double helix and to impart other structural features, such as an altered groove width and disruption of the normal spine of hydration. DNA containing DAP may or may not be recognized by restriction endonucleases; RNA containing DAP may not engage in normal splicing. The DAP.T pair affects the local flexibility of DNA and impedes the interaction with helix bending proteins. By providing a non-canonical hydrogen bond donor in the minor groove and/or blocking access to the floor of that groove it strongly affects interactions with small molecules such as antibiotics and anticancer drugs. Examples which illustrate altered recognition of nucleotide sequences in DAP containing DNA are presented: changed sites of cutting by bleomycin, photocleavage by uranyl nitrate and footprinting with mithramycin. Using DNA in which both A-->DAP and G-->Inosine substitutions have been made it is possible to assess precisely the role of the purine 2-amino group in ligand-DNA recognition. PMID- 9742231 TI - The activity binding to the termination region of several pol III genes represents a separate entity and is distinct from a novel component enhancing U6 snRNA transcription. AB - Human TFIIIC1, a basal transcription factor essentially required for expression of all pol III genes, exerts its function without primarily binding to DNA. We report here the purification of a termination site binding activity (TBA) which was initially described to be contained in fractions designated as TFIIIC0. TBA specifically and strongly binds to the termination region of pol III genes with internal promoters and can be completely separated from TFIIIC1and a TFIIIC1related activity (TFIIIC1-like), proving that DNA-binding of TBA is independent of these latter activites. Although TBA is not essentially required for, it strongly stimulates pol III transcription from intragenic promoters. This stimulation strictly depends on the presence of TFIIIC1and is not observed in conjunction with TFIIIC1-like. We further present the identification of a novel activity, TFIIIU, which is also contained in crude fractions of TFIIIC0. TFIIIU can be separated from TBA by further purification and is essentially involved in transcription of the mammalian U6 gene. TFIIIU cannot be substituted for by any of the established U6 transcription factors and thus represents a novel U6 transcription factor. PMID- 9742232 TI - Molecular and biochemical characterization of new X-ray-sensitive hamster cell mutants defective in Ku80. AB - Ku, a heterodimer of approximately 70 and approximately 80 kDa subunits, is a nuclear protein that binds to double-stranded DNA ends and is a component of the DNA-dependent protein kinase (DNA-PK). Cell lines defective in Ku80 belong to group XRCC5 of ionizing radiation-sensitive mutants. Five new independent Chinese hamster cell mutants, XR-V10B, XR-V11B, XR-V12B, XR-V13B and XR-V16B, that belong to this group were isolated. To shed light on the nature of the defect in Ku80, the molecular and biochemical characteristics of these mutants were examined. All mutants, except XR-V12B, express Ku80 mRNA, but no Ku80 protein could clearly be detected by immunoblot analysis in any of them. DNA sequence analysis of the Ku80 cDNA from these mutants showed a deletion of 252 bp in XR-V10B; a 6 bp deletion that results in a new amino acid residue at position 107 and the loss of two amino acid residues at positions 108 and 109 in XR-V11B; a missense mutation resulting in a substitution of Cys for Tyr at position 114 in XR-V13B; and two missense mutations in XR-V16B, resulting in a substitution of Met for Val at position 331 and Arg for Gly at position 354. All these mutations cause a similar, 5-7-fold, increase in X-ray sensitivity in comparison to wild-type cells, and a complete lack of DNA-end binding and DNA-PK activities. This indicates that all these mutations lead to loss of the Ku80 function due to instability of the defective protein. PMID- 9742233 TI - In vitro method for the generation of protein libraries using PCR amplification of a single DNA molecule and coupled transcription/translation. AB - A novel in vitro method for the generation of a protein library has been developed using the polymerase chain reaction (PCR) amplification of a single DNA molecule followed by in vitro coupled transcription/translation. DNA template encoding green fluorescent protein of a jellyfish Aequorea victoria was extensively diluted to one molecule per well, and then amplified by a total of 80 cycles of PCR with nested primers. The exact number of origins in the amplified DNA fragment was then estimated by directly sequencing a part of the fragment, at which an individual template molecule was marked by PCR with a primer containing three randomized bases. Since the sequences obtained in 91 independent amplifications were diversified statistically, each amplified fragment was likely originated from a single DNA molecule. In addition, the amplified fragments served as a template for in vitro coupled transcription/translation using T7 RNA polymerase and Escherichia coli S30 extract. These results suggest that the library obtained by the PCR amplification of a single DNA molecule diluted from a variety of DNA pools is potentially useful in high-throughput generation of protein libraries. PMID- 9742234 TI - DNA complexes obtained with the integron integrase IntI1 at the attI1 site. AB - Integrons are genetic elements that are able to capture genes by a site-specific recombination mechanism. Integrons contain a gene coding for a lambda-like integrase that carries out site-specific recombination by interacting with two different target sites; the attI site and the palindromic sequence attC (59 base element). Cassette integrations usually involve the attI site, while cassette excisions use attC . Therefore, the integrase should bind both sites to cleave DNA and perform site-specific recombination reactions. We have used purified maltose-binding protein fused with the integrase (MBP-IntI1) and native IntI1 protein and gel retardation assays with fragments containing the complete and partial attI1 site to show formation of four complexes in this region. Chemical modification of specific nucleotides within the attI1 site was used to investigate their interference with binding of the integrase protein. We attribute IntI1 specific binding to four regions in the attI1 site and a GTTA consensus sequence is found in three of the four regions. Interference by modified guanine and thymine residues in the DNA major groove and adenine residues in the minor groove were observed, indicating that the integrase interacts with both sides of the helix. Binding of IntI1 to attC is also discussed. PMID- 9742235 TI - Transfer RNA mimicry among tymoviral genomic RNAs ranges from highly efficient to vestigial. AB - Three tRNA-associated properties of a representative set of tymoviral RNAs have been quantitatively assessed using higher plant (wheat germ) proteins: aminoacylation, EF-1alpha*GTP binding, and 3'-adenylation of 3'-CC forms of the RNAs by CTP, ATP:tRNA nucleotidyltransferase. The RNAs fall into three classes differing in the extent of tRNA mimicry. Turnip yellow mosaic (TYMV) and kennedya yellow mosaic virus RNAs had activities in all three properties similar to those of a higher plant tRNAValtranscript, and thus are remarkable tRNA mimics. Although the isolated approximately 83 nt long tRNA-like structures showed high activity in these assays, in the case of TYMV, the 6318 nt long TYMV RNA was an even better substrate for valylation. Eggplant mosaic virus RNA, which has a differently constructed acceptor stem pseudoknot, differed from the above tymoviral RNAs in binding more weakly to EF-1alpha*GTP. Erysimum latent virus RNA, which lacks an identifiable anticodon domain, could not be valylated and had very low 3'-adenylation activity. The range of tRNA mimicry within the tymovirus genus thus ranges from extremely highly developed to minimal. The implications on the role of the tRNA mimicry in viral biology are discussed. PMID- 9742236 TI - mtDNA replicative potential remains constant during ageing: polymerase gamma activity does not correlate with age related cytochrome oxidase activity decline in platelets. AB - Progressive age-related oxidative phosphorylation (OxPhos) decline is well known in human tissues. Depletion of mitochondrial DNA (mtDNA) causes OxPhos defects in patients with myopathic syndromes and deficient mtDNA replication has been observed in cells cultured from patients with mitochondrial disease. Patients undergoing treatment for AIDS develop OxPhos defects via mtDNA depletion resulting from inhibition of mtDNA polymerase gamma (Polgamma) by 2'-deoxy 3' azido thymidine. These findings by others give rise to a possible link between mtDNA replication and bioenergetic decline in disease and during ageing. We have designed an in vitro assay for Polgamma function in small tissue samples to explore this possible link. Platelet homogenate Polgamma showed an activity with a K m of 150 microM (dTTP), a V max of 11.8 pmol/min/mg, inhibited (41% inhibition; 50 microM) by ethidium bromide. Determination of several storage characteristics showed that platelets were a convenient source of Polgamma for assay. Polgamma activity in 45 subjects did not coincide with significant age related decline (P<0.002; P) observed in cytochrome oxidase (CytOx) activity or with citrate synthase activity. Of the activities studied, the only significant age-wise variation was a 24% CytOx deficiency in elderly (>50; n = 19) compared to young (<51; n = 24) individuals (P<0.01; t). These results suggest a maintenance of total cellular mtDNA Polgamma processive levels during ageing, largely independent of total cellular bioenergetic status or mitochondrial number/density. The processive component of Polgamma is therefore unlikely to make a major contribution to age-related bioenergetic activity decline. This does not, however, preclude the possibility that transient periods of inhibition at crucial points of the cell cycle or development may augment existing intracellular deficiencies. The assay described here greatly facilitates study of Polgamma activity in patients with conditions involving mtDNA depletion or rearrangement. PMID- 9742237 TI - Cross-species aminoacylation of tRNA with a long variable arm between Escherichia coli and Saccharomyces cerevisiae. AB - Prokaryotes have three amino acid-specific class II tRNAs that possess a characteristic long variable arm, tRNASer, tRNALeuand tRNATyr, while eukaryotes have only two, tRNASerand tRNALeu. Because of such a phylogenetic divergence in the composition of tRNA, the class II tRNA system is a good candidate for studying how the tRNA recognition manner has evolved in association with the evolution of tRNA. We report here a cross-species aminoacylation study of the class II tRNAs, showing the unilateral aminoacylation specificity between Escherichia coli and a yeast, Saccharomyces cerevisiae. Both SerRS and LeuRS from E.coli were unable to aminoacylate yeast class II tRNAs; in contrast, the yeast counterparts were able to aminoacylate E.coli class II tRNAs. Yeast seryl-tRNA synthetase was able to aminoacylate not only E.coli tRNASerbut also tRNALeuand tRNATyr, and yeast LeuRS was able to aminoacylate not only E.coli tRNALeubut also tRNATyr. These results indicate that the recognition manner of class II tRNA, especially the discrimination strategy of each aminoacyl-tRNA synthetase against noncognate class II tRNAs, is significantly divergent between E.coli and yeast. This difference is thought to be due mainly to the different composition of class II tRNAs in E.coli and yeast. PMID- 9742238 TI - The double-stranded RNA-binding protein X1rbpa promotes RNA strand annealing. AB - RNA-annealing activity is a common feature of several RNA-binding proteins. The Xenopus RNA-binding protein X1rbpa is composed of three tandemly arranged double stranded RNA-binding domains (dsRBDs) but lacks any other catalytic or functional domains, therefore making the assessment of biological functions of this protein rather difficult. Here we show that full-length X1rbpa but also isolated dsRBDs from this protein can facilitate RNA strand annealing. RNA annealing can be efficiently inhibited by heparin. However, dsRBDs with a neutral pI still promote strand annealing, suggesting that charged residues within the dsRBD are important for strand annealing. Additionally, mutant versions of the dsRBD, unable to bind dsRNA in northwestern assays, were tested. Of these, some show RNA-annealing activity while others fail to do so, indicating that RNA annealing and dsRNA binding are separable functions. Our data, together with the previously reported association of the protein with most cellular RNAs, suggests an RNA chaperone like function of X1rbpa. PMID- 9742240 TI - Repair of DNA strand gaps and nicks containing 3'-phosphate and 5'-hydroxyl termini by purified mammalian enzymes. AB - A putative role for mammalian polynucleotide kinases that possess both 5' phosphotransferase and 3'-phosphatase activity is the restoration of DNA strand breaks with 5'-hydroxyl termini or 3'-phosphate termini, or both, to a form that supports the subsequent action of DNA repair polymerases and DNA ligases, i.e. 5' phosphate and 3'-hydroxyl termini. To further assess this possibility, we compared the activity of the 3'-phosphatase of purified calf thymus polynucleotide kinase towards a variety of substrates. The rate of removal of 3' phosphate groups from nicked or short (1 nt) gapped sites in double-stranded DNA was observed to be similar to that of 3'-phosphate groups from single-stranded substrates. Thus this activity of polynucleotide kinase does not appear to be influenced by steric accessibility of the phosphate group. We subsequently demonstrated that the concerted reactions of polynucleotide kinase and purified human DNA ligase I could efficiently repair DNA nicks possessing 3'-phosphate and 5'-hydroxyl termini, and similarly the combination of these two enzymes together with purified rat DNA polymerase beta could seal a strand break with a 1 nt gap. With a substrate containing a nick bounded by 3'- and 5'-OH termini, the rate of gap filling by polymerase beta was significantly enhanced in the presence of polynucleotide kinase and ATP, indicating the positive influence of 5' phosphorylation. The reaction was further enhanced by addition of DNA ligase I to the reaction mixture. This is due, at least in part, to an enhancement by DNA ligase I of the rate of 5'-phosphorylation catalyzed by polynucleotide kinase. PMID- 9742241 TI - Single-tube nested competitive PCR with homologous competitor for quantitation of DNA target sequences: theoretical description of heteroduplex formation, evaluation of sensitivity, precision and linear range of the method. AB - Competitive PCR is a frequently used technique for quantitation of DNA and mRNA. However, the application of the most favourable homologous mutated competitors is impeded by the formation of heteroduplex molecules which complicates the data evaluation and may lead to quantitation errors. Moreover, in most cases a single quantitation of an unknown sample requires multiple competitive reactions for identification of the equivalence point. In the present study, a highly efficient and reliable method as well as the underlying theoretical model is described. The mathematical solutions of this model provide the basis for single-tube quantitation using a homologous mutated competitor. For quantitation of Human Papilloma Virus 16-DNA, it is shown that single tube quantitations using simple PAGE separation and video evaluation for signal analysis permit linear detection within more than two orders of magnitude. In addition, repeated single-tube competitive PCRs exhibited good precision (average standard deviation 5%), even if carried out as nested high cycle PCR for quantitation of low abundant sequences (intraassay sensitivity <2 x 10(2) copies). This evaluation method can be applied to any DNA separation and detection method which is capable of resolving the heteroduplex fraction from both homoduplex molecules. PMID- 9742239 TI - The influence of 3TC-resistance mutations E89G and M184V in the human immunodeficiency virus reverse transcriptase on mispair extension efficiency. AB - Two nucleoside analog resistance mutations in HIV-1 reverse transcriptase (RT), E89G and M184V, were previously shown to increase the dNTP insertion fidelity of HIV-1 RT. However, forward mutation assays using a lacZ alpha reporter gene have revealed a lack of impact on the overall error rate of these variants. In an effort to investigate the basis for this discrepancy, we have examined whether the increases in misinsertion fidelity observed for E89G and M184V RTs are accompanied by an increase in mispair extension fidelity. The relative efficiencies with which the wild type, E89G, M184V and M184V/E89G HIV-1 RTs extend model template-primer duplexes containing 3'-OH terminal mismatches were measured. The calculated efficiencies of mispair extension ( f ext) were, in general, not significantly decreased from the wild type HIV-1 RT. In fact, the efficiency of extension from one of the mispaired primer-template duplexes was significantly increased for two of the mutants tested. These results suggest that amino acid substitutions that increase the fidelity of dNTP insertion do not necessarily increase misextension fidelity, and that the decreased misextension fidelity may counterbalance the increases in misinsertion fidelity observed for E89G and M184V RTs. PMID- 9742243 TI - AT-hook motifs identified in a wide variety of DNA-binding proteins. AB - The AT-hook is a small DNA-binding protein motif which was first described in the high mobility group non-histone chromosomal protein HMG-I(Y). Since its discovery, this motif has been observed in other DNA-binding proteins from a wide range of organisms. Using pattern searches and position-dependent matrices, we have extracted the AT-hook motifs present in a non-redundant protein sequence database. We have classified these motifs into three types according to their sequence similarity and have found that they are prevalent in many eukaryotic nuclear proteins in single or multiple copies. Furthermore, AT-hook motifs are frequently associated with known functional domains seen in chromatin proteins and in DNA-binding proteins (e.g. histone folds, homeodomains and zinc fingers). In general, it appears that the AT-hook motif is an auxiliary protein motif cooperating with other DNA-binding activities and facilitating changes in the structure of the DNA either as a polypeptide on its own [e.g. HMG-I(Y)] or as part of a multidomain protein [e.g. Swi2p in Saccharomyces cerevisiae or HRX (ALL 1) in Homo sapiens]. It is most interesting that this motif seems to be quite specific to known or predicted chromosomal/DNA-binding proteins, suggesting that it may act as a versatile minor groove tether. PMID- 9742242 TI - Characterization of two intein homing endonucleases encoded in the DNA polymerase gene of Pyrococcus kodakaraensis strain KOD1. AB - Two intein endonucleases, denoted PI- Pko I and PI- Pko II, in the DNA polymerase gene of the hyperthermophilic archaeon Pyrococcus kodakaraensis KOD1 were expressed in Escherichia coli and the recombinant endonucleases were characterized. Both endonucleases were thermostable and cleaved their intein-less DNA sequences leaving four base 3'-hydroxyl overhangs. PI-Pko I exhibited 22 times higher specific activity than PI-Pko II and the activity of PI-Pko II was enhanced at higher potassium ion concentrations (1 M). Recognition sequences were also determined using synthetic oligonucleotides inserted into plasmid pUC19. It was shown that DNA sequences of 19 and 16 bp are needed for cleavage by PI-Pko I and PI-Pko II, respectively. PI-Pko II could cleave the downstream junction region between intein-encoding and mature DNA polymerase regions and cleavage by PI-Pko II could be detected even when chromosomal DNA of P.kodakaraensis KOD1 was used as substrate. Therefore, it is suggested that these endonucleases are switching endonucleases whose function lies in the rearrangement of chromosomal DNA. PMID- 9742244 TI - Homogeneous rate of degradation of nuclear DNA during apoptosis. AB - DNA fragmentation during apoptosis is characterized by endonucleolytic cleavage of chromosomal DNA into an oligonucleosomal ladder. To determine if actively transcribed genes are more susceptible to cleavage during apoptosis than non transcribed genes, the rate of fragmentation of differentially expressed genes was measured in B-lymphocyte hybridoma cells. Five genes were studied based on their transcriptional activity and/or nuclear localization, and mitochondrial DNA was assayed as a negative control for apoptotic fragmentation. Apoptosis was induced in the hybridoma cells by ultraviolet light, and DNA was prepared at multiple time points after ultraviolet irradiation. Degradation into an oligonucleosomal ladder appeared as early as 2 h after treatment, showing that fragmentation is rapidly activated in hybridoma cells. The DNA was then digested with restriction enzymes, separated by gel electrophoresis and hybridized with the gene-specific probes for Southern blot analyses. Loss of gene-specific signals was measured by quantitation of autoradiographs. The results show all of the nuclear genes were degraded at the same rate regardless of their transcriptional status or nuclear localization. The data suggest that once the cell activates its destruction program, nuclear DNA is rapidly degraded in a homogeneous manner. PMID- 9742245 TI - Expression profiling of single cells using 3 prime end amplification (TPEA) PCR. AB - The ability to relate the physiological status of individual cells to the complement of genes they express is limited by current methodological approaches for performing these analyses. We report here the development of a robust and reproducible method for amplifying 3' sequences of mRNA derived from single cells and demonstrate that the amplified cDNA, derived from individual human lymphoblastoma cells, can be used for the expression profiling of up to 40 different genes per cell. In addition, we show that 3 prime end amplification (TPEA) PCR can be used to enable the detection of both high and low abundance mRNA species in samples harvested from live neurons in rat brain slices. This procedure will facilitate the study of complex tissue function at the cellular level. PMID- 9742246 TI - An apyrimidinic site kinks DNA and triggers incision by endonuclease VII of phage T4. AB - Apurinic/apyrimidinic lesions (AP-sites) occur frequently in DNA, generated by physically and chemically induced or spontaneous loss of bases. Repair mechanisms have evolved in organisms to deal efficiently with AP-sites by first incising the DNA at the lesion, followed by excision and resynthesis of the damaged strand. Here we report that endonuclease VII (endo VII) of phage T4, which was originally classified as a debranching and Holliday structure resolving enzyme, also recognizes AP-sites with high efficiency. The enzyme cleaves both strands of double-stranded DNA in a stepwise fashion a few nucleotides 3' of the lesion. In a search for a recognition signal shared by all known endo VII substrates, kinking of DNA has earlier been suggested as such a signal. In support of this hypothesis, we demonstrate here that AP-sites induce distinct kinks in synthetic oligonucleotides allowing efficient intramolecular ring closure by ligation. PMID- 9742247 TI - Analysis of the subunit assembly of the typeIC restriction-modification enzyme EcoR124I. AB - Type I restriction-modification (R-M) enzymes are composed of three different subunits, of which HsdS determines DNA specificity, HsdM is responsible for DNA methylation and HsdR is required for restriction. The HsdM and HsdS subunits can also form an independent DNA methyltransferase with a subunit stoichiometry of M2S1. We found that the purified Eco R124I R-M enzyme was a mixture of two species as detected by the presence of two differently migrating specific DNA protein complexes in a gel retardation assay. An analysis of protein subunits isolated from the complexes indicated that the larger species had a stoichiometry of R2M2S1and the smaller species had a stoichiometry of R1M2S1. In vitro analysis of subunit assembly revealed that while binding of the first HsdR subunit to the M2S1complex was very tight, the second HsdR subunit was bound weakly and it dissociated from the R1M2S1complex with an apparent K d of approximately 2.4 x 10(-7) M. Functional assays have shown that only the R2M2S1complex is capable of DNA cleavage, however, the R1M2S1complex retains ATPase activity. The relevance of this situation is discussed in terms of the regulation of restriction activity in vivo upon conjugative transfer of a plasmid-born R-M system into an unmodified host cell. PMID- 9742248 TI - Different cleavage specificities of RNases III from Rhodobacter capsulatus and Escherichia coli. AB - 23S rRNA in Rhodobacter capsulatus shows endoribonuclease III (RNase III) dependent fragmentation in vivo at a unique extra stem-loop extending from position 1271 to 1331. RNase III is a double strand (ds)-specific endoribonuclease. This substrate preference is mediated by a double-stranded RNA binding domain (dsRBD) within the protein. Although a certain degree of double strandedness is a prerequisite, the question arises what structural features exactly make this extra stem-loop an RNase III cleavage site, distinguishing it from the plethora of stem-loops in 23S rRNA? We used RNase III purified from R.capsulatus and Escherichia coli, respectively, together with well known substrates for E.coli RNase III and RNA substrates derived from the special cleavage site in R.capsulatus 23S rRNA to study the interaction between the Rhodobacter enzyme and the fragmentation site. Although both enzymes are very similar in their amino acid sequence, they exhibit significant differences in binding and cleavage of these in vitro substrates. PMID- 9742250 TI - Mechanistic studies on the DNA linking activity of Epstein-Barr nuclear antigen 1. AB - The DNA replication, plasmid segregation and transactivation functions of Epstein Barr nuclear antigen 1 (EBNA1) require the binding of EBNA1 to specific DNA recognition sites in the two non-contiguous functional elements of the Epstein Barr virus latent origin of replication, oriP . EBNA1 molecules bound to these elements interact with each other resulting in the formation of looped individual DNA molecules and multiply linked DNA molecules. We have developed a glycerol gradient sedimentation assay suitable for quantitative analysis of the DNA linking activity of EBNA1 and used it to investigate the contribution of EBNA1 residues to the linking interaction and the mechanism of the interaction. Using overlapping internal deletion mutants, we found that two regions of EBNA1 can cause DNA linking, amino acids 40-100 and 327-377, but that the stabilities of the linked complexes formed by the two regions differ dramatically; only complexes formed through the latter region are stable to glycerol gradient sedimentation analysis. Mechanistic studies using EBNA1 in combination with GAL4 EBNA1 fusion proteins showed that linking interactions mediated by residues 327 377 are homotypic. Our results also suggest that only the DNA-bound form of EBNA1 participates in the protein-protein interactions seen in DNA linking. PMID- 9742249 TI - Modulation of plasmid DNA methylation and expression in zebrafish embryos. AB - Gene expression is under the influence of DNA methylation and assembly of chromatin structure. This paper reports the modulation of transgene expression in zebrafish embryos by altering DNA methylation with 5-azacytidine and heterochromatin formation with sodium butyrate, an inhibitor of histone deacetylation. A CMV promoter-luciferase fusion gene construct (pCMVL) microinjected into zebrafish eggs becomes gradually methylated during development, starting at approximately 12 h post-injection. When methylated in vitro by Hpa II methylase prior to injection, the construct is rapidly demethylated in vivo before being de novo methylated. Demethylation is independent of DNA replication, indicating that it is an active DNA repair process. Demethylating activity has been characterized in zebrafish embryo nuclear extracts, in which this activity is heat-labile, sensitive to protease and RNase and requires ATP hydrolysis. Demethylating activity in vitro is dependent on the developmental stage of the embryo from which extracts are prepared. In vivo , luciferase transcripts are detected prior to de novo plasmid methylation. Furthermore, incubation of pCMVL-injected embryos with 5-azacytidine or butyrate immediately after injection inhibits plasmid methylation and extends the period of luciferase expression. When applied after de novo methylation has occurred, both inhibitors prevent methylation of newly replicated DNA and promote transgene expression. These data suggest that methylation of the injected construct during early development induces repression of the transgene, perhaps by converting the construct to a repressive chromatin structure. PMID- 9742251 TI - Drosophila RpS3a, a novel Minute gene situated between the segment polarity genescubitus interruptus and dTCF. AB - Genetic analysis of the small chromosome 4 of Drosophila has been hampered by the virtual lack of recombination. The segment polarity gene cubitus interruptus (ci) maps to the most intensively studied locus on this chromosome. Up to four complementation groups have been found to be associated with ci. We and others have recently characterized a second segment polarity gene, dTCF or pan, 12 kb upstream of ci, in a head-to-head configuration. During the course of these studies we identified a transcription unit in the intergenic region. We report here the cloning of cDNAs from this transcription unit, which encode the Drosophila homologue of the human ribosomal protein S3a (RpS3a). The RpS3a gene is expressed ubiquitously and throughout development. A Minute allele, M(4)101, linked tightly to ci, was found to harbour an integration of a Doc retroposon in the promotor region of RpS3a. Thus, like other Minute loci, M(4)101 encodes a component of the protein synthesis machinery. These data further unravel the complex genetics surrounding the ci and dTCF loci. PMID- 9742252 TI - Identification of fragments of human transcripts froma defined chromosomal region: representational difference analysis of somatic cell hybrids. AB - We have tested representational difference analysis of cDNAs from somatic cell hybrids as a means to directly isolate expressed sequences derived from a defined chromosomal region. To this end, the hamster-human somatic cell hybrid Q1Z, which carries Xq28 as the only human chromosomal fragment, was used as Tester and the parental hamster cell line Y21 as Driver. After two rounds of subtraction, two major products of 510 and 307 bp were obtained, derived from the highly expressed human Xq28-derived QM gene and from a hamster repeat sequence strongly up regulated in Q1Z, respectively. In a second subtraction experiment these fragments were added to the driver, to prevent their reappearance. After three rounds of subtraction a more complex difference product was obtained. Of 26 different fragments analysed, 12 fragments were derived from Xq28-derived genes, 10 of which were from known genes. One fragment was derived from a hamster gene strongly up-regulated in Q1Z. These results demonstrate that cDNA RDA can be used to isolate gene fragments from defined chromosomal regions and that suppression with major products, derived from highly expressed genes, is advantageous to isolate larger number of fragments, presumably derived from rarer transcripts. PMID- 9742253 TI - Concordance analysis of microbial genomes. AB - The set of proteins which are conserved across families of microbes contain important targets of new anti-microbial agents. We have developed a simple and efficient computational tool which determines concordances of putative gene products that show sets of proteins conserved across one set of user specified genomes and not present in another set of user specified genomes. The thresholds and the homology scoring criterion are selectable to allow the user to decide the stringency of the homologies. The system uses a relational database to store protein coding regions from different genomes, and to store the results of a complete comparison of all sequences against all sequences using the FASTA program. Using Web technology, the display of all the related proteins for a given sequence and calculation of multiple sequence alignments (using CLUSTALW) can be performed with the click of a button. The current database holds 97 365 sequences from 19 complete or partial genomes and 8798905 FASTA comparison results. A example concordance is presented which demonstrates that the target of the quinolone antibiotics could have been identified using this tool. PMID- 9742254 TI - Mutational analysis of a transcriptional activation region of the VP16 protein of herpes simplex virus. AB - The VP16 protein of herpes simplex virus is a potent transcriptional activator of the viral immediate early genes. The transcriptional activation region of VP16 can be divided into two functional subregions, here designated VP16N (comprising amino acids 413-456) and VP16C (amino acids 450-490). Assays of VP16C mutants resulting from both random and alanine-scanning mutagenesis indicated that the sidechains of three phenylalanines (at positions 473, 475 and 479) and one acidic residue (glutamate 476) are important for transcriptional activation. Aromatic and bulky hydrophobic amino acids were effective substitutes for each of the three Phe residues, whereas replacement with smaller or polar amino acids resulted in loss of transcriptional function. In contrast, many changes were tolerated for Glu476, including bulky hydrophobic and basic amino acids, indicating that the negative charge at this position contributes little to the function of this subregion. Similar relative activities for most of the mutants were observed in yeast and in mammalian cells, indicating that the structural requirements for this activation region are comparable in these two species. These results reinforce the hypothesis that bulky hydrophobic residues, not acidic residues, are most critical for the activity of this 'acidic' transcriptional activation region. PMID- 9742255 TI - Isolation of genes negatively or positively co-expressed with human recombination activating gene 1 (RAG1) by differential display PCR (DD RT-PCR). AB - Differential display PCR (DD RT-PCR) has been extensively used for analysis of differential gene expression, but continues to be hampered by technical limitations that impair its effectiveness. In order to isolate novel genes co expressing with human RAG1, we have developed an effective, multi-tiered screening/purification approach which effectively complements the standard DD RT PCR methodology. In 'primary' screens, standard DD RT-PCR was used, detecting 22 reproducible differentially expressed amplicons between clonally related cell variants with differential constitutive expression of RAG mRNAs. 'Secondary' screens used differential display (DD) amplicons as probes in low and high stringency northern blotting. Eight of 22 independent DD amplicons detected nine independent differentially expressed transcripts. 'Tertiary' screens used reconfirmed amplicons as probes in northern analysis of multiple RAG-and RAG+sources. Reconfirmed DD amplicons detected six independent RAG co-expressing transcripts. All DD amplicons reconfirmed by northern blot were a heterogeneous mixture of cDNAs, necessitating further purification to isolate single cDNAs prior to subcloning and sequencing. To effectively select the appropriate cDNAs from DD amplicons, we excised and eluted the cDNA(s) directly from regions of prior northern blots in which differentially expressed transcripts were detected. Sequences of six purified cDNA clones specifically detecting RAG co-expressing transcripts included matches to portions of the human RAG2 and BSAP regions and to four novel partial cDNAs (three with homologies to human ESTs). Overall, our results also suggest that even when using clonally related variants from the same cell line in addition to all appropriate internal controls previously reported, further screening and purification steps are still required in order to efficiently and specifically isolate differentially expressed genes by DD RT-PCR. PMID- 9742256 TI - Preferential cleavage in pre-ribosomal RNA byprotein B23 endoribonuclease. AB - Protein B23 is an abundant nucleolar protein and a putative ribosome assembly factor which possesses an intrinsic ribonuclease activity. In the current work, the effects of RNA sequence and secondary structure on the cleavage preference by protein B23 were studied. Protein B23 ribonuclease preferentially cleaved the single-stranded homopolymers poly(A), poly(U) and poly(C). However, double stranded co-polymers and poly(G) were resistant to cleavage. No base specificity was observed with an oligoribonucleotide substrate. The action of protein B23 ribonuclease on different regions of pre-rRNA was studied using transcripts synthesized in vitro from cloned rDNA segments. Although no specific cleavages were detected in transcripts containing sequences from the 5' external transcribed spacer or the first internal transcribed spacer, the enzyme preferentially cleaved the second internal transcribed spacer (ITS2) approximately 250 nt downstream from the 3'-end of 5.8S rRNA. Preferential cleavage was retained when the transcript was extended by 100 nt at the 3'-end, but abolished in a transcript lacking this cleavage site. Furthermore, this site was not susceptible to cleavage by RNase A or RNase T1. These results, in conjunction with the sub-nucleolar localization of the protein with elements of the processing machinery, suggest that the protein B23 endoribonuclease could play a role in pre-rRNA processing in ITS2. PMID- 9742258 TI - Purification of plasmids by triplex affinity interaction. AB - Production of pharmaceutical grade plasmid DNA is an important issue in gene therapy. We developed a method for affinity purification of plasmids by triple helix interaction. This method is based on sequence-specific binding of an oligonucleotide immobilized on a large pore chromatography support to a target sequence on the plasmid. Using design criteria derived from thermodynamic data, we produced a 15mer target sequence which binds strongly to the affinity support under mildly acidic conditions. Plasmid DNA was purified from clarified Escherichia coli lysate by incubation with the affinity beads at pH 5.0 and high NaCl concentration. After extensive washing of the beads, purified plasmid DNA was eluted with alkaline buffer. The purified plasmid showed no RNA or cell DNA contamination in HPLC analysis and total protein concentration was reduced considerably. Due to its mechanical stability and porosity this support can be used in a continuous affinity purification process, which has a high potential for scale up. PMID- 9742257 TI - In vivo selection for intronic splicing signals from a randomized pool. AB - Retroviruses utilize balanced splicing to express multiple proteins from a single primary transcript. A number of cis -acting signals help maintain this balance, including the branch point sequence (BPS), polypyrimidine tract (PPyT) and sequences within the downstream exon. In general, regulated splicing requires weak splicing signals and we have previously shown the same requirement for the simple retrovirus, avian sarcoma virus (ASV). Here we take advantage of the requirement for balanced splicing in retroviral replication to examine the sequence constraints of an intronic splicing element. Selection for replication competence makes it possible to amplify and identify functional sequences from a pool of all possible sequences. In this report we examine the role of pyrimidines within the PPyT. Our results provide in vivo confirmation that the functional strength of a PPyT is related to its length and uridine content and that the PPyT plays a role in the second step of the splicing reaction. We also show that the minimal distance between the 3'-splice site and the BPS in this system is 16 nt. With modification, the selection system described here can be used to examine the sequence constraints of other exonic or intronic splicing elements in vivo . PMID- 9742259 TI - Gene amplification and transcriptional upregulation of the sarco/endoplasmic reticulum Ca2+ transport ATPase in thapsigargin-resistant hamster smooth muscle cells. AB - We have selected a series of cell lines from the parental Syrian hamster smooth muscle cell line DDT1-MF2that are resistant to thapsigargin (TG), a specific inhibitor of the sarcoplasmic/endoplasmic reticulum Ca2+transport ATPases (SERCAs). Cells were selected for resistance to TG in the presence or absence of cyclosporin (CSA), which is a competitive inhibitor of the multidrug transporter p-glycoprotein (pgp). Since TG is a known substrate for pgp, selection for TG resistance was carried out in the presence of CSA in an attempt to minimize the contribution of pgp, and to identify the potential range of adaptive responses of the SERCA pump itself, during the development of the TG-resistant phenotype. Irrespective of whether the selection is carried out in the presence or absence of CSA, pgp is overexpressed in the TG-resistant DDT1-MF2cells. SERCA protein is also overproduced in the TG-resistant cell lines, which occurs through one of several mechanisms. Included among these, is amplification of the SERCA gene and enhanced transcription of the gene. Enhanced transcription is observed only upon long-term selection and occurs through the SERCA gene proximal promoter elements. Although SERCA transcription in wild-type cells is dependent upon the -284 to -72 bp region of the SERCA promoter, the TG-resistant cells utilize both the -284 to 72 bp and the -72 to +80 bp promoter regions for enhanced SERCA transcription. That is, additional elements within the -72 to +80 bp region are recruited in the TG-resistant cells to allow for increased SERCA expression. A post transcriptional step may also be recruited by the TG-resistant cells in their overall strategy to produce increased amounts of the SERCA protein. These studies demonstrate that the DDT1-MF2cells can utilize different mechanisms which lead to increased levels of SERCA protein as the cells adapt to inhibition of the ATPase by TG. PMID- 9742260 TI - Isolation of developmentally regulated genes by differential display screening of cDNA libraries. AB - mRNA differential display RT-PCR has been extensively used for the isolation of genes differentially expressed between RNA populations. We have assessed its utility for the identification of developmentally regulated genes in plasmid cDNA libraries derived from individual tissues dissected from early mouse embryos. Using plasmid Southern blot hybridisation as a secondary screen, we are able to identify such genes and show by whole-mount in situ hybridisation that their expression pattern is that expected from the differential display profile. PMID- 9742261 TI - Congenital diaphragmatic hernia. A cause of persistent pulmonary hypertension of the newborn which lacks an effective therapy. AB - Congenital diaphragmatic hernia (CDH) is still an unsolved problem. A disease which was, for a long time, thought to be merely a hole in the diaphragm appears today to be an intriguing malformation with a poorly understood pathogenesis and a complex pathophysiology. CDH results in various degrees of pulmonary hypoplasia and severe persistent pulmonary hypertension of the newborn. Despite antenatal ultrasound diagnosis and continuous improvement in neonatal intensive care, these features could not be overcome, and the overall mortality rate in CDH is still reaching 50%. Experimental works during the past 20 years suggest that CDH is a disease of impaired lung development associated with, but not caused by, a structural defect of the diaphragm. Furthermore, there is increasing evidence that the lung in CDH is not only small but that there are numerous disorders (e.g. surfactant deficiency, decreased anti-oxidant activity, increased vascular reactivity with decreased nitric oxide and increased endothelin 1 activity, and left heart hypoplasia) which may be associated with impaired lung development. Although antenatal diagnosis of CDH is feasible by ultrasound, there is no reliable predictor of pulmonary hypoplasia, the main prognostic factor in CDH. Whilst modern therapeutic strategies such as high-frequency oscillatory ventilation, exogenous surfactant or inhaled nitric oxide may be beneficial in selected subjects, most newborns with hypoplastic lungs will not survive despite these postnatal therapies. Perhaps these newborns would benefit from antenatal treatment directed at altering lung growth early in utero to minimize pulmonary hypoplasia. Therefore, research is needed to elucidate the aetiology and pathogenesis of CDH. Knowledge about the cellular control of proliferation, differentiation and programmed cell death (apoptosis) in the organogenesis should clarify our understanding of these processes and allow us to develop drugs that stimulate lung growth or even correct the anatomical defect. Furthermore, early and reliable assessment of prognosis for fetuses with CDH at risk of death will become increasingly important in the identification of fetuses most likely to benefit from antenatal therapies and may eventually lead to decreased mortality. PMID- 9742263 TI - Improved oxygenation following adenosine infusion in persistent pulmonary hypertension of the newborn. AB - Six consecutive cases of persistent pulmonary hypertension of the newborn (PPHN) were treated with adenosine following failure of conventional therapy, excluding inhaled nitric oxide. A rise in arterial PO2 >20 mm Hg occurred in 5 of 6 cases within 30 min of commencing adenosine infusion. Individual maximal increases in PaO2 ranged from 31 to 131 mm Hg. Three neonates survived and 3 died. Amongst deaths, intensive support was withdrawn in a preterm neonate due to severe arthrogryposis/pulmonary hypoplasia. Of the remaining 2, the improvement in oxygenation persisted until death occurred from causes unrelated to adenosine. Side effects related to adenosine (bradycardia, hypotension, prolonged bleeding time) did not occur. Due to its ease of availability, administration and extremely short half-life, adenosine may be an important therapeutic option in PPHN. PMID- 9742262 TI - Indications of coagulation and/or fibrinolytic system activation in healthy and sick very-low-birth-weight neonates. AB - A combined hemostatic defect consisting of a reduction in certain procoagulants, anticoagulants (antithrombin III-ATIII-, protein C-PC-) and components of the fibrinolytic system (plasminogen-Plg-) was demonstrated in very-low-birth-weight infants (VLBW <1,500 g) with gestational age 26-32 weeks. Sixteen of them were healthy, 28 were suffering from RDS and 24 from septicemia. The hemostatic defect was more profound in the RDS group, nevertheless increased TAT (thrombin + ATIII complex) and/or PAP values (plasmin + a2-antiplasmin complex) was a more frequent finding in the septicemic group of infants (91.8 vs. 17.9%). Moderate-to-severe thrombocytopenia was detected in a higher percentage in the septicemic (70.8%) than in the RDS group (50%), and increased D-dimers were demonstrated in 34.8 and 28.6% of the infants, respectively. Elevated TAT or PAP values were not always associated with gross coagulation abnormalities, and advanced disseminated intravascular coagulation (DIC) was only documented in 16.7% of the septicemic and 7.1% of the RDS infants. None of the VLBW neonates presented with clinical evidence of thrombosis, although hemorrhagic manifestations were apparent in 34.8 and 14.3% of the neonates with septicemia or RDS, respectively, mainly due to DIC or severe thrombocytopenia. In conclusion, increased TAT and/or PAP values are good indicators of the in vivo activation of the hemostatic system, but still their impact on sick neonates morbidity and mortality remains unknown. PMID- 9742264 TI - Adaptation in neonatology of the once-daily concept of aminoglycoside administration: evaluation of a dosing chart for amikacin in an intensive care unit. AB - BACKGROUND: The bactericidal efficacy of aminoglycosides is directly related to peak serum concentration (Cmax), particularly the first one. Transitory high concentrations of aminoglycosides do not result in such a high drug uptake by renal and cochlear tissues because of the saturation of cell binding sites. These observations have led to the concept that less frequent administration of relatively larger doses of aminoglycosides would be of interest in treating infectious diseases. OBJECTIVE: Prospective evaluation of a dosing chart of amikacin (Ak) in high-risk neonates suspected of infection within the first 2 days of life. This dosing chart was based on a previous pharmacokinetic population study published elsewhere, treated accordingly to the new once-daily concept of aminoglycoside administration. STUDY DESIGN: One hundred and seventy seven neonates (69 females and 108 males; mean gestational age (GA +/-SD: 33.6 +/ 4.1 weeks (W) received Ak regimen dosage according to the following dosing chart: Group (Gr) 1a GA <28 W: 20 mg/kg/42 h; Gr 1b GA 28 /= 37 W: 15.5 mg/kg/24 h. In case of asphyxia, hypoxic episode and intercourse treatment with indomethacin, the interval was systemically increased by 6 h whatever the GA groups. The mean duration time of Ak treatment (+/- 1 SD) was 5.00 +/- 2.01 days (range 2-13). Ak serum concentrations 1 h after completion of 30 min infusion (C1h), and successive Ak serum concentrations just before next administration depending on the difference of interval between each group (so defined minimum serum concentration (Cmin)), were determined in each neonate. Creatininemia during the fist postnatal weeks was used as an index of glomerular filtration rate; brainstem auditory evoked potentials (BEAPs) were used in 139 babies when reaching a postconceptional age of >/= 36 weeks to assess possible ototoxicity, and were compared to values from a group of term and a group of preterm babies, previously defined as our reference control groups. RESULTS: At day 1 of treatment, there was no correlation between the Ak C1hS and the GA at birth (mean 27.8 +/- 5.21 microgram/ml (+/- 1 SD); median 28; r = -0.003; range 10-40). In the same way, there was no correlation between the first Ak CminS and the GA at birth (mean 3.7 +/- 2.0 microgram/ml (+/- 1 SD); median 3.0; r = -0.33; range 0-10). The lack of correlation between these first observed C1hS and CminS and the GA at birth suggests the validity of our previous established dose regimen recommendations. Analyzing the data between groups, the mean value +/- 1 SD of Ak C1hS at day 1 of treatment was not significantly different (p > 0.05). Concerning the first Ak CminS, a significant difference (p < 0.01) was only observed when comparing groups 1a, 1b and 2 to group 4. However, this significant difference disappeared when comparing the successive next Ak CminS between groups while each interval remained the same, suggesting a positive postnatal maturation of the renal clearance. In the same way, creatininemia showed a significant and normal decrease (p < 0.01) in each group during the first postnatal weeks. Threshold values of BEAPs at 30 dB showed no significant difference (p > 0.05) between the treated groups (preterm group and term group) and the corresponding control groups. While the primary aim of the study was not to test the bactericidal efficacy of this new regimen, the recovery was excellent in 37 babies with proven or highly suspected infectious disease, except in 1 of them who died from septic shock (group B Streptococcus). After 5 years of using this kind of Ak administration in the unit, minimal inhibitory concentration profiles tested in 43 successive bacterial strains collected from inborn patients remained adequate. (ABSTRACT TRUNCATED) PMID- 9742265 TI - The effect of recombinant TGFalpha, human milk, and human milk macrophage media on gut epithelial proliferation is decreased in the presence of a neutralizing TGFalpha antibody. AB - OBJECTIVE: An in vitro model was devised to compare the relative effects of recombinant transforming growth factor-alpha (TGFalpha), aqueous human milk, and human milk macrophage (HMM) medium on human fetal small intestinal cell (FHs-74) proliferation. METHODS: Recombinant TGFalpha at increasing concentrations (range 0.01-1,000 ng/ml media), the aqueous fraction of human milk (AHM), or HMM medium was added to FHs-74 cells in the presence or absence of a neutralizing TGFalpha antibody (1 microgram/ml medium). At 24 h, cell proliferation was measured and expressed as percent control. The experimental variables were (1) activators of cell growth (TGFalpha, AHM, and HMM medium); (2) increasing concentrations of TGFalpha, and (3) neutralizing antibody to TGFalpha. The dependent variable for all experiments was cell proliferation. RESULTS: Significant effects for growth stimulators and TGFalpha concentration as measured by cell proliferation were found. Specifically, there was a dose-dependent effect of TGFalpha on cell proliferation to the 5-ng/ml concentration, with a plateau reached in cell proliferation at higher concentrations. The stimulatory effect of TGFalpha was decreased in the presence of TGFalpha antibody (mean +/- SD 22 +/- 7. 1% decline, p < 0.001). In the presence of TGFalpha antibody, there was a 25 +/- 3.1% decline in HM-stimulated growth (p < 0.004), and a 27.6 +/- 3.2% decline in HMM medium stimulated growth (p < 0.001). CONCLUSIONS: Neutralization of recombinant TGFalpha and that present in human milk and HMM medium by TGFalpha antibody led to a consistent decrease in in vitro human fetal small intestine epithelial proliferation without affecting cell viability. These results support the hypothesis that TGFalpha, whether derived from human recombinant sources, human milk or HMM medium has a measurable, trophic effect on in vitro human gut epithelial cells. PMID- 9742266 TI - The fetal glycoprotein, fetuin, counteracts ill-effects of the bacterial endotoxin, lipopolysaccharide, in pregnancy. AB - We show that fetuin, a fetal glycoprotein present in fetal plasma in concentrations substantially higher than in the adult, can exert a protective function against ill-effects of the bacterial endotoxin, lipopolysaccharide (LPS), in pregnancy. The concentration of fetuin in plasma of pregnant rats (E17 gestation) was artificially increased by repeated intraperitoneal administration of this protein. This was followed by an injection of a standard dose of LPS, a dose that produced in control animals (no additional fetuin) nearly 50% abortions and 40% maternal deaths. None of the females exposed to increased fetuin died following LPS injection. Nine out of 10 multigravida animals carried their pregnancy to full term but all primagravidas aborted. PMID- 9742267 TI - Correlation of near infrared spectroscopy cerebral blood flow estimations and microsphere quantitations in newborn piglets. AB - We compared cerebral blood flow (CBF) estimated using transmission mode near infrared spectroscopy (NIRS) and a modification of the Fick principle with CBF quantitations by radioactive microspheres (MSs) in newborn piglets. Thirteen piglets were studied during steady state, ischemia, and during two reflow periods. NIRS and MS flows were not significantly different during any measurement period. NIRS flows were compared to total brain blood flows and to regional brain blood flows quantitated with MSs and correlated best with temporal cortical flows. Linear regression analysis of the NIRS flows plotted against MS quantitated temporal cortical flows showed r = 0.71. Thus, CBFs obtained with NIRS were not significantly different from, showed the same directional changes, and correlated acceptably with flows quantitated by MSs. PMID- 9742268 TI - Increase in the renal damage induced by paracetamol in rats exposed to ethanol translactationally. AB - Administration of ethanol (8%) or acetone (1%) to nursing dams in the drinking water, for 10 days, increased the nephrotoxicity of paracetamol (APAP) in the 14 day-old lactating offspring. The percentage of proximal tubular cells with evidence of necrotic damage in male rats was higher in those animals that received APAP (500 mg/kg, i.p.) and whose nursing rats were exposed to ethanol (25. 0 +/- 8.4%) or acetone (17.2 +/- 1.2%), than in the group treated with APAP alone (10.6 +/- 1.6%). The activity of urinary N-acetylglucuronidase was also significantly higher in the rats exposed translactationally to ethanol or acetone than in animals treated with the APAP alone. Nephrotoxicity showed a sexual dimorphic pattern with a higher toxicity in male than in female rats. The percentage of necrotic tubules in the male rats not exposed to inductor was 10.6 +/- 1.6%, and in female rats 5.0 +/- 1.4% (p < 0. 05). Animals exposed to ethanol or acetone and treated with APAP showed less weight gain than the group treated only with APAP. Our results suggest that renal toxicity is enhanced in the nursing animals that were exposed, via maternal milk, to ethanol or acetone (inductors of cytochrome P4502E1), than in the control animals. This circumstance may be relevant in alcoholic women while they are lactating. PMID- 9742269 TI - Addiction, intoxication, criminal law and criminal justice: an introduction. PMID- 9742270 TI - Intoxication, the law and criminal responsibility--a sparkling cocktail at times: the case studies of Canada and Germany. AB - The combination of intoxication and criminal responsibility has been a problem field for legal theory and practice for quite some time. While it has been argued in certain contexts that intoxication reduces or denies criminal responsibility, elsewhere it has been reasoned that intoxicated offenders should be held as (or even more so) legally responsible as sober ones. But even in legal systems where the criminal responsibility of intoxicated offenders is emphasized, legal theory and practice are confronted with the challenge of converting such values into workable jurisprudence, since many intoxicated offenders naturally lack one of the key premises for responsibility for a criminal act, namely mens rea. This article compares the very different legal philosophies and practices that have evolved around the issue of intoxication and criminal responsibility in Canada and Germany. While the Canadian system has long and in a variety of ways tried to reconcile the inherent tensions between the principles of legal culpability and the intent to punish intoxicated offenders in material law, the German system has produced a set of legal tools that allow for a pragmatic and ends-oriented approach. This article concludes that the evolution and profile of these legal schemes is likely linked to the cultural status of alcohol and drinking in the respective system context. PMID- 9742271 TI - Intoxication and criminal responsibility in Dutch criminal Law. AB - This article deals with the question in how far an offence committed in the Netherlands under the influence of alcohol or other drugs can be imputed to the offender. Unlike many other countries the Dutch Penal Code does not contain specific provisions with respect to the criminal liability of addicted or intoxicated offenders. In principle, they are held responsible for their offences, even when the dolus or culpa is absent at the moment they commit their offence. Doctrine and jurisprudence found this liability on the principle of 'culpa/dolus in causa', by accepting an anterior dolus or culpa, which is situated at the moment the offender takes alcohol or other drugs. As is shown in this article, the - nondogmatic - interpretation of this culpa in causa doctrine leaves hardly any space for a claim to impunity. PMID- 9742272 TI - Psychiatric criteria of legal responsibility after the consumption of alcohol: the German situation. AB - Psychiatry deals with addiction but it is not concerned very much about the symptomatology of acute alcohol intoxication. This causes problems in connection with the legal system: intoxication is the most frequent cause of diminished legal responsibility in Germany, and judges want to have a system to assess the grade of intoxication as simply and reliably as possible. So they found the blood alcohol concentration (BAC) to be a safe indication. But the BAC has various effects not only on different people but also on the same person in different situations: other factors are influential, i.e. alcohol habituation and physical condition, personality, mood and situation. Therefore it is necessary to describe the typical syndromes of intoxication and their dependency on BAC. PMID- 9742273 TI - Recent changes in Danish law on drugs and drug offences. AB - The article recounts changes in Danish Drug Law and Enforcement since the beginning of the 1990s and relates them to general trends in Danish criminal policy during the period. In addition to the implementation of EU directives, e.g. on money laundering and growth hormones, legislation has been passed to curb conspicuous dealing of drugs in the streets of Copenhagen. This part of the legislation is seen as a reaction to public fears and reactions to visible aliens dealing in drugs in a conspicuous way, albeit in minor quantities. The legal changes imply a considerable rise in penalties for repeated dealings in minor quantities and easier access to deportation of aliens. The latter has been criticized as potential violation of the human rights of aliens. This and other recent changes in criminal law and related legislation is seen as an indication of politicians' concerns with voters' anxieties, possibly at the edge of moral panics. PMID- 9742274 TI - Promotion of evaluation as a strategy towards an effective substance prevention. AB - A growing number of European initiatives focusing on the evaluation and monitoring of substance abuse prevention is observed. At the beginning of the 90s a team from the Institute of Psychiatry and Neurology in Warsaw, Poland, began a project designed for the promotion of evaluation in the field of substance abuse prevention. The project included outcome studies on popular school-based programs and development of teaching materials and publications to popularize the concept and methods of evaluation. As results of the project several outcome studies were completed, several scales for evaluating alcohol prevention programs were developed, a number of articles and a handbook on evaluation were published. PMID- 9742275 TI - L-Methadone and D,L-methadone in methadone maintenance treatment: a comparison of therapeutic effectiveness and plasma concentrations. AB - The clinical effectiveness of l-methadone maintenance treatment (LMMT) carried out using d,l-methadone or l-methadone have been compared with ambulatory heroin dependent subjects. A total of 40 heroin-dependent subjects, previously maintained on l-methadone in Frankfurt am Main, were divided into two groups under randomised double-blind conditions and received either an equivalent dose of l-methadone as d,l-methadone or remained on the previous l-methadone treatment. Requests for a change in the dose of d,l-methadone and l-methadone were recorded, urine samples for determination of illicit drug use were collected and the individual level of opiate craving was determined over a 22-day observation period. There was no significant difference between the two groups in the number requests for a dose change (dose increase <10%). However, there was a significant increase in heroin use in the group which continued to receive l methadone. Although there was less variability in opiate craving in the group receiving d,l-methadone, the mean intensity of opiate craving did not differ between the two groups. The mean l-methadone dose:l-methadone plasma concentration ratio, an index of the bioavailability of l-methadone in individual subjects, showed no significant change when the treatment was changed to d,l methadone. The mean d-methadone:l-methadone plasma concentration ratio was 1.17. There was no significant difference between these ratios for day 15 and day 22. The mean l-methadone:EDDP plasma concentration ratio in the l-methadone group was 22.2 and the d,l-methadone:EDDP plasma concentration ratio was 18.4 . The plasma EDDP concentration in the d,l-methadone group increased 3-fold after starting treatment with d, l-methadone. These findings suggest that d,l-methadone can be used in methadone maintenance treatment of heroin-dependent subjects but that further studies are required to evaluate pharmacokinetic interactions between methadone enantiomers. PMID- 9742276 TI - Diagnosis of cochlear Meniere's disease with electrocochleography. AB - The existence of cochlear Meniere's disease, once considered a variant of classic Meniere's disease but without vertigo, has been questioned due to lack of objective evidence that endolymphatic hydrops is involved with the disease process. Transtympanic electrocochleography (TT ECoG) has emerged as a useful tool for electrophysiologic monitoring of the inner ear, and is especially valuable in assessing endolymphatic hydrops. A retrospective chart review was performed to identify those patients with a diagnosis consistent with cochlear Meniere's disease in order to determine the presence or absence of endolymphatic hydrops using TT ECoG. A total of 7 patients were identified with at least a 2 year follow-up. Using established norms for the summating potential to action potential ratio with click stimulus, 67% of the ears examined demonstrated values consistent with endolymphatic hydrops. Fluctuating aural pressure and tinnitus were present in all patients and medical therapy of diuretics and salt restriction seemed to stabilize or improve the condition in about 80% of the patients. Theoretical considerations are discussed, and a case history of 1 of the study patients is presented to illustrate a typical example of this variant of Meniere's disease. PMID- 9742277 TI - The effect of hemodialysis on hearing using pure-tone audiometry and distortion product otoacoustic emissions. AB - Patients under treatment of hemodialysis (HD) frequently exhibit some degree of sensorineural hearing loss. Fifteen subjects and 10 controls were tested by using pure-tone audiometry (PTA) and distortion-product otoacoustic emissions (DPOAEs) before and after a HD treatment. Other parameters (blood pressure, body weight, blood chemistries) were also evaluated before and after HD: The purpose of this study was to determine the acute effect of HD on hearing level by measuring PTA and DPOAE before and after one HD session. The results from PTA and DPOAE testings showed that hearing was unaffected by HD. However, all 15 subjects revealed significantly poorer hearing, especially in the higher frequencies, compared to that of the controls (p < 0.0001). It was concluded then that HD is a safe treatment, and that the sensorineural hearing loss in these patients may be attributed to the preexisting renal disease. PMID- 9742278 TI - Effect of acoustic trauma on cytochrome oxidase activity in stria vascularis. AB - The present study was undertaken to determine the role of metabolic disturbance in noise-induced hearing loss by histochemical studies of cytochrome oxidase activity. Adult normal albino guinea pigs were used. The experimental animals were exposed to broad-band noise at 105 dB SPL for 24 h. The control animals were not exposed to the noise. The thresholds of the auditory brainstem response (ABR) of all guinea pigs were measured 3 times: before noise exposure, 1 day and 1 month later. The difference between the ABR thresholds before and after noise exposure was statistically significant. Vibratome sections of decalcified cochleae of the noise-exposed (n = 8) and control groups (n = 4) were incubated with Spector's medium and embedded with Epon. Thin sections (2 microm) and ultrathin sections (100 nm) were cut to observe cytochrome oxidase activity in the stria vascularis under light and electron microscopes, respectively. A decreased activity of cytochrome oxidase was consistently shown in the normal appearing stria vascularis of most noise-exposed ears. Acoustic trauma has an adverse effect on cytochrome oxidase activity in the stria vascularis as well as on hearing. A decrease in the activity of cytochrome oxidase implicates that metabolic damage may play a role in noise-induced hearing loss. PMID- 9742279 TI - Bilateral cholesterol granulomas of the temporal bone. AB - Cholesterol granulomas of the temporal bone without clinical symptoms of chronic otitis media may result from an indolent inflammatory process caused by a congenitally blocked group of air cells. A unique case of giant bilateral cholesterol granulomas of the temporal bone is presented to support this theory. Management allowed bilateral surgical removal with hearing preservation. The differentiation between cholesterol granuloma, giant cholesterol cyst and other lesions of the temporal bone is discussed. The presumed pathogenesis of this condition is reviewed. PMID- 9742280 TI - Titanium and glass-ionomer cement as ossicular replacement materials: biocompatibility results after implantation in the rabbit. AB - The middle ear poses unique challenges when finding suitable materials for ossicular reconstruction, primarily because of its link to the external environment via the eustachian tube, which leads to a greater potential for exposure to infectious agents. In this animal study, the biocompatibilities of titanium and glass-ionomer cement were assessed in the middle ear of the rabbit after being implanted as total ossicular replacement prostheses (TORPs) or as free pins. Animals were sacrificed after 28, 84, 168, 336, or 504 days or 2 years, and a cutting saw technique was used to prepare slides for light microscopy. Slides were examined for mucosal coverage and any sign of foreign body reaction. Both materials showed good acceptance in the middle ear. After 28 days, the TORPs were covered by middle ear mucosa. As expected, it took a longer time (up to 504 days) to cover the free implants. An interesting finding was the growth of new bone on both the surface of the titanium implants and the glass ionomer prostheses. The results of this animal study indicate that both titanium and glass-ionomer cement are favorable materials for ossicular replacement prostheses. PMID- 9742281 TI - The effect of Epstein-Barr virus gene BHRF1 expression on radioresistance of nasopharyngeal carcinoma cells. AB - In order to investigate the effect of the expression of Epstein-Barr virus gene BHRF1 on the apoptotic resistance of nasopharyngeal carcinoma cells to radiation, a highly expressing vector for BHRF1 was constructed and transfected into the nasopharyngeal carcinoma cell line CNE2. Then, the biologic alterations of the cells were tested after 60Co radiation. The results showed that, in the BHRF1 expressing cells, the apoptotic index was far lower than in the control groups after 60Co radiation, and cells recovered faster from the radiation, with a higher cell-proliferative rate, stronger ability of colony formation and tumor development in nude mice than that in the control groups. Given the functional homology of BHRF1 with bcl-2, our data indicate that BHRF1 expression could prohibit nasopharyngeal carcinoma cellular apoptosis caused by radiation and in this way contribute to oncogenic transformation. PMID- 9742282 TI - Changes in nasal reactivity in patients with rhinitis medicamentosa after treatment with fluticasone propionate and placebo nasal spray. AB - AIM OF THE STUDY: To study the changes in nasal reactivity in patients with rhinitis medicamentosa during treatment with placebo or fluticasone propionate, in order to better understand the mechanisms of nasal congestion in such patients. STUDY DESIGN: A parallel, double-blind study. Twenty patients with rhinitis medicamentosa were randomized to either placebo or fluticasone treatment during 14 days. MATERIAL AND METHODS: Nasal mucosa reactivity was studied with a histamine challenge model using three concentrations of histamine to challenge the nasal mucosa (1, 2 and 4 mg histamine/ml). Recordings of the nasal mucosa response were made 5 min after each challenge, using rhinostereometry and acoustic rhinometry, before and after the period of treatment. RESULTS: The fluticasone group had a significantly increased histamine sensitivity after treatment, unlike the placebo group who had an unchanged or slightly decreased histamine sensitivity after treatment. CONCLUSIONS: The results of this study support the theory that the nasal obstruction in rhinitis medicamentosa is due to interstitial oedema rather than to vasodilatation. On the first day of vasoconstrictor withdrawal, the inferior concha was congested and oedematous with a limited capacity to respond to histamine challenge. However, after 14 days of treatment with a corticosteroid nasal spray, the oedema was reduced and the increase in histamine sensitivity reflected the persistence of nasal hyperrreactivity. In the placebo group, histamine sensitivity remains unchanged with the measuring technique we used. This probably indicates that oedema was still present after treatment. PMID- 9742283 TI - Lectin expression during wound healing of the rabbit sinus mucosa: a study of regenerating epithelium and early polyp formation. AB - Lectin expression during wound healing of the rabbit sinus mucosa was examined. Positive UEA-I staining was evident on squamous or cuboidal as well as columnar regenerating epithelial cells (RE cells). PNA staining of columnar RE cells first became evident after neuraminidase treatment, while squamous or cuboidal RE cells stained positively with PNA alone. Fucosylation within RE cells thus occurred from a relatively early period, and sialylation followed at later stages. Ingrowing epithelial cells of early polyp formation stained negatively with UEA I, indicating that unfucosylated RE cells may represent aberrant cellular behavior. We concluded that these patterns of lectin staining indicate a functional maturation as well as an integration of regenerating mucosa. PMID- 9742284 TI - Frey's syndrome following submandibular gland excision: an unusual postoperative complication. AB - Gustatory sweating and flushing, or Frey's syndrome, is a fairly common complication following surgery or injury to the parotid gland and is thought to be caused by aberrant nerve regeneration. A similar condition has been reported in the literature following surgery to the submandibular region. Since this was first described in 1934, only 7 subsequent cases of submandibular sweating and flushing have been reported. We present a case of a 52-year-old female who underwent excision of the left submandibular gland as a result of chronic sialadenitis. Twelve months following surgery, symptoms indicative of Frey's syndrome were experienced in the operative region. A review of the aetiology and treatment of the condition is described. PMID- 9742286 TI - Reply PMID- 9742285 TI - Far-advanced otosclerosis. PMID- 9742287 TI - Enhancement of morphine antinociception by ibogaine and noribogaine in morphine tolerant mice. AB - The effects of ibogaine, an alkaloid isolated form the bark of the African shrub, Tabernathe iboga, and noribogaine, a metabolite of ibogaine, on morphine antinociception were determined in male Swiss-Webster mice. Mice were rendered tolerant to morphine by implanting them with a pellet containing 25 mg of morphine base for 3 days. Placebo pellet-implanted mice served as controls. The antinociception of morphine (10 mg/kg, s.c.) was determined alone or in combination with an appropriate dose of ibogaine or noribogaine. Tolerance to morphine developed as a result of morphine pellet implantation as evidenced by decreased antinociceptive response to morphine. Both ibogaine and noribogaine dose-dependently enhanced morphine antinociception in morphine-tolerant but not in morphine-naive mice. It is concluded that ibogaine and noribogaine enhance morphine antinociception in morphine-tolerant mice. PMID- 9742288 TI - Anticonvulsant properties of 1,4-benzodiazepine derivatives in amygdaloid-kindled seizures and their chemical structure-related anticonvulsant action. AB - The effects of 14 different 1,4-benzodiazepines on amygdaloid-kindled seizures and their chemical structure-related anticonvulsive actions were studied. The prophylactic effects of 1, 4-benzodiazepines on amygdaloid-kindled seizures were also examined. Male Wistar strain rats were used in this study. Rats were anesthetized with pentobarbital sodium (35 mg/kg i.p.) and bipolar electrodes were implanted into the right amygdala. The stimulating parameters were 1 ms pulse duration, 60 Hz frequency and a 1 s duration at an intensity just sufficient to induce afterdischarge (AD). All the 1,4-benzodiazepines depressed both seizure stage and AD duration of amygdaloid-kindled seizures. Of the 1, 4 benzodiazepines, prazepam, flutoprazepam and flurazepam with a long alkyl chain at position 1 were less effective than the drugs having a hydrogen or methyl group at the same position. Nitrazepam, nimetazepam, flunitrazepam and clonazepam which have a nitro group at position 7 showed more potent antiepileptic activity than the drugs with a chloro group. Certain 1,4-benzodiazepines caused inhibition of the development of amygdaloid-kindled seizures. The existence of a hydrogen or methyl group at position 1 and a nitro group at position 7 is important for exhibiting potent anticonvulsant activity in amygdaloid-kindled seizures. Introduction of an oxygen group at position 2 is also necessary for high activity. 1,4-benzodiazepines had not only therapeutic but also prophylactic effects on amygdaloid-kindled seizures. PMID- 9742289 TI - Thyroid accumulation and adverse effects of imipramine and desipramine in rats after long-term administration. AB - The respective adverse effects of imipramine and desipramine on serum thyroid hormone levels and their accumulation in thyroid were investigated in male Wistar rats. Two groups of 30 rats were gavaged for 4 weeks with 30 mg/kg/day imipramine hydrochloride (IMI) or desipramine hydrochloride (DESI), while the control group (12 rats) received the arabic gum vehicle only. In the IMI-treated group, the serum thyroxine (T4) level significantly decreased (by 13%) and IMI and its metabolite DESI were accumulated in the thyroid, as pointed out by mean thyroid to-serum concentration ratios close to 12 and 8, respectively. In the DESI treated group, the mean thyroid-to-serum concentration ratio of the drug was close to 14, and significant decreases in both serum T4 (-20%) and triiodothyronine serum levels (-14%) were found. The accumulation of antidepressant drugs in the thyroid was more pronounced and the thyroid serum levels were even lower after DESI administration than after IMI administration. These results are in favour of an antithyroid action of IMI and DESI due to the formation of a complex in the thyroid between molecular iodine and the drugs or metabolites. PMID- 9742290 TI - Lisuride acts at multiple sites to induce ocular hypotension and mydriasis. AB - Topically unilaterally applied lisuride caused dose-related lowering of intraocular pressure in ipsilateral (treated) but not in contralateral eyes of normal rabbits. The ocular hypotensive response induced by lisuride was antagonized by pretreatment with metoclopramide, a dopamine receptor antagonist, and was partially reduced by local sympathetic denervation. In contrast to the unilateral effect on intraocular pressure, lisuride caused mydriasis in both eyes. Mydriasis was of greater magnitude and more sustained in normal eyes compared to sympathetically denervated eyes. Additional in vivo experiments demonstrated that lisuride caused dose-related suppression of neuronally initiated contractions of cat nictitating membrane. In in vitro experiments lisuride caused dose-related inhibition of norepinephrine release from isolated rabbit iris-ciliary bodies. Pretreatment with Bay K 8644, a calcium channel activator, did not attenuate lisuride-induced inhibition of norepinephrine release in isolated rabbit iris-ciliary bodies. Because lisuride pretreatment caused no change in isoproterenol-stimulated cAMP accumulation in isolated iris ciliary bodies, suppression of adenylate cyclase was unlikely. It is concluded that the ocular hypotensive effect of lisuride results, in part, from activation of prejunctional dopaminergic receptors on peripheral sympathetic nerves in the anterior segment of the eye but may also involve antagonism on peripheral postjunctional alpha1 adrenoceptors as well. Bilateral increases in pupil diameter antagonized by metoclopramide suggest a stimulatory action of lisuride on dopamine receptors in the central nervous system. PMID- 9742291 TI - Effects of morin on an experimental model of acute colitis in rats. AB - The flavonoid morin was tested for anti-inflammatory activity in trinitrobenzenesulfonic acid (TNBS)-induced rat colitis. Rats were pretreated orally with several doses of the flavonoid (5, 10, 25, 100 and 200 mg/kg) 48, 24 and 1 h before and 24 h after colitis induction and examined for colonic damage 48 h after colitis induction. Colonic inflammation was characterized by diffuse hemorrhagic necrosis of the mucosa, bowel wall thickening, impairment of fluid absorption, increase in myeloperoxidase (MPO) activity, enhanced leukotriene B4 (LTB4) synthesis, glutathione depletion and increased levels of malonyldialdehyde (MDA). Morin treatment, at doses ranging from 10 to 200 mg/kg, significantly reduced colonic macroscopic damage. This beneficial effect was also confirmed by inhibition of colonic MPO activity. Several mechanisms may contribute to the protective effect exerted by morin. First, inhibition of colonic LTB4 synthesis is a common feature for all the active doses of the flavonoid. Second, the antioxidant properties of morin, which partially prevented colonic glutathione depletion (at doses of 10 and 25 mg/kg) or inhibited colonic MDA production (at doses of 100 and 200 mg/kg), can collaborate in preventing TNBS-induced inflammation. PMID- 9742292 TI - Inhibitory effects of isradipine on spontaneous and oxytocin- and carbachol stimulated contractions of rat myometrium. AB - Our aim was to investigate the effect of isradipine, a second-generation calcium channel blocker, on spontaneous and oxytocin- and carbachol-stimulated contractions of myometrium isolated from nonpregnant and pregnant rats. Amplitude, frequency, duration and integrated area of spontaneous and oxytocin- and carbachol-stimulated contractions of nonpregnant and pregnant rat myometrium were compared before and after the treatment with isradipine (10(-6) to 10(-4) mol/l). Isradipine inhibited contractions of myometrial strips isolated from nonpregnant and pregnant rats in a concentration-dependent manner, with a significant effect on the amplitude (10(-5) to 10(-4) mol/l) and integrated area (3 x 10(-6) to 10(-4) mol/l) of spontaneous and oxytocin- and carbachol stimulated contractions. Isradipine induced similar concentration-dependent effects on the frequency and duration of spontaneous and carbachol-stimulated contractions at higher concentrations (3 x 10(-5) to 10(-4) mol/l) but had no effect on frequency and duration of oxytocin-stimulated contractions. Isradipine appears to be an effective relaxant in rat myometrium. This effect of isradipine may become therapeutically advantageous in clinical application for preterm labor. PMID- 9742293 TI - Possible contribution of leukotrienes in the arrhythmogenic effects of digoxin on isolated guinea-pig hearts. AB - The effect of a leukotriene D4 (LTD4) receptor antagonist, L-648,051, was investigated in digoxin-induced cardiac toxicity in isolated guinea-pig hearts (Langendorff preparation). Digoxin infusion (25 microg.ml-1, 0.5 ml.min-1) increased perfusion pressure and contractile force initially, but decreased them later. The onset of first ventricular premature beats (VPBs) matched the increase phase, but the decrease phase was accompanied by ventricular tachycardia (VT) and fibrillation (VF). In the presence of L-648,051 (5 micromol. l-1), the initial phase was similar to that observed with digoxin alone, but the marked reduction was inhibited. This drug increased the concentration of digoxin required for VBSs and cardiac arrest, but it could not prevent the formation of VT and VF. The duration of VT was significantly decreased by L-648,051. It is concluded that the leukotriene receptor antagonist might have beneficial effects on digoxin-induced arrhythmias. Whether this effect depends on direct or indirect actions is uncertain. PMID- 9742294 TI - The Chirality Medal Award 1997 bestowed upon Professor Ryoji Noyori, Nagoya University, Japan. PMID- 9742295 TI - On the helical paths traced by rising bubbles. PMID- 9742296 TI - Folate and its various ramifications. PMID- 9742297 TI - Prevention of injuries to children and adolescents. AB - Injury prevention is one of the most important preventive health challenges for pediatricians worldwide. A science of injury control has developed. Matching a child's skill and development age is needed for anticipatory guidance. Poor children living in rural areas are at greatest risk and require continuous reinforcement. Family function relates closely to injuries and recovery from injury. Prevention involves education, legislation, environmental modification, and engineering techniques. PMID- 9742298 TI - The development and disorders of sleep. AB - Sleep-related problems are extremely frequent sources of parental concern and question. Many childhood sleep difficulties are simply minor behavioral problems, whereas other sleep changes represent serious disorders. Increased knowledge about the development and disorders of sleep has enormous value in making accurate diagnoses and effective interventions. Overviews are presented on common and important sleep disorders in infancy, childhood, and adolescence, including night walking, partial arousal, sleep-disordered breathing, narcolepsy, and sleep loss secondary to late and erratic schedules. This chapter also draws attention to the close links between the regulation of sleep and the control of attention, emotion, and behavior. The evidence that many children and adolescents obtain less than optimal amounts of sleep raises a number of important questions about rising rates of behavioral and emotional problems. PMID- 9742299 TI - Melatonin: the dark force. AB - Although the pineal gland was described 2,300 years ago, its functions remained obscure and productive research was limited until 1958, when Lerner and associates defined melatonin. In 1965 Wurtman and Axelrod advanced the "melatonin hypothesis," according to which the pineal gland acts as a transducer responding to changes in circumambient light by changing its rates of melatonin output. Sites and mechanisms of melatonin action are still poorly understood. Two consistent effects are the induction of sleep and an antigonadotropic influence on reproductive structure and behavior. The former is demonstrable and clinically useful in human subjects; the latter has been shown in birds, rodents, and sheep. Alteration of skin color by the contraction of melanophores was effected by pineal extracts before the discovery of melatonin. This phenomenon, seen in reptiles, amphibians, and fish, has received little recent attention. Areas of greater interest and potential importance include the antimitotic effects of melatonin on some types of tumor cells in culture and the apparent in vivo protection of immunocompetent lymphocytes during chronic stress, which reduces the functional capacity of lymphocytes in control rodents. Clinical application of the antimitotic and immunosupportive properties of melatonin seems likely in the near future. Unfortunately, this innocent molecule has been touted in two recent books and many advertisements as an aphrodisiac, rejuvenator, protector against disease, and general wonder-worker. Because interest in melatonin is high, all physicians can expect questions and may have use for the information provided in this review. PMID- 9742300 TI - Adolescent homosexuality. AB - Homosexuality has existed in all civilizations, but societal disapproval and cultural taboos have negatively influenced its recognition. A significant percentage of youths identify themselves as homosexual, and even more experience sex with the same sex or are confused about sexual feelings. A unifying etiological theory attributes the expression of sexual orientation to genes that shape the central nervous system's development, organization, and structure via prenatal sex steroids. Environmental factors may influence the expression of genetic potential. Several models of psychosocial development describe initial stages of awareness and confusion about same-sex attractions, followed by acknowledgement of homosexuality, disclosure to others, and eventual integration of sexual identity into a comprehensive sense of self. Stressors related to isolation, stigma, and violence may predispose homosexual adolescents to impaired social, emotional, and physical health, resulting in depression and suicide, school problems, substance abuse, running away eating disorders, risky sexual behavior, and illegal conduct. As with all adolescents, the overall goals in the care of homosexual youth are to promote normal adolescent development, social and emotional well-being, and physical health. A comprehensive, multidisciplinary approach is required to address medical, mental health, and psychosocial issues within the context of the adolescents' community and culture. PMID- 9742301 TI - Food allergy: a clinical approach. AB - Adverse reactions to foods are common and may be life threatening. Many of the reactions are immunologic (allergic), and the types of these reactions are discussed. The most common type is IgA mediated. Clinical manifestations are multiple. Systematic investigation is usually rapid and cost-effective and may be life saving. PMID- 9742302 TI - An updated view of the value of taurine in infant nutrition. PMID- 9742303 TI - Immaturity of gastrointestinal host defense in newborns and gastrointestinal disease states. AB - Not only is the gastrointestinal tract the largest immune organ in the body, but it also contains one of the most important and interesting immunologic compartments. Host protection against pathogens and injurious agents by the gastrointestinal tract is essential for an individual's survival. The intestinal mucosal immune system, which is linked with other mucosal surfaces and together represents the common mucosal immune system, prevents the passage of potentially harmful antigens and pathogens into the systemic circulation of the host. In a healthy host, antigens crossing the mucosal barrier in physiologic quantities evoke the appropriate immune response, which includes polymeric IgA antibody production to the antigen and systemic tolerance. PMID- 9742304 TI - The placenta and its significance in neonatal outcome. AB - The placenta should not be overlooked as a source of much valuable diagnostic information. Close evaluation of the placenta and its attached membranes may reveal further information. Essential data may be obtained from pathologic examination, and if questions exist, specimens should be retained with proper care. PMID- 9742305 TI - Genetics of congenital heart disease: strategies. AB - Congenital malformations of the heart are the most common of all birth defects. Traditionally, a multifactorial model combining genetic predisposition with environmental influence has been cited as the cause of greater than 90% of heart disease. This may be too broad inasmuch as linkage analysis combined with the explosion of information derived from the Human Genome Project has allowed the identification of genetic defects for many types of acquired and congenital diseases of the heart. This review provides a summary of cardiac conditions for which genetic etiologies are apparent, as well as an introduction to some basic clinical genetic concepts. Despite significant advances, it is important to remember that we are still very early in our understanding of the relationship of genotype to phenotype and that the clinical implications of the genetic defects identified are incompletely understood and have only begun to be studied. PMID- 9742306 TI - Human glucose transporters. AB - Concentrative and facilitative glucose transporters are responsible for the movement of glucose across the plasma membrane of human cells. Defects in concentrative glucose transporters cause renal glycosuria and glucose-galactose malabsorption. Alterations in facilitative glucose transporters explain the newly discovered syndrome of low CNS glucose in the presence of normal blood sugar, causing seizures and developmental delay. Defects in other facilitate glucose transporters also help explain Fanconi-Bickel syndrome, glycogen storage disease type, Id, and non-insulin-dependent diabetes mellitus. PMID- 9742307 TI - Pathobiology and clinical significance of molecular genetic findings in childhood tumors. AB - Molecular genetic determinants of malignant diseases in children are providing insights into the mechanisms of malignancies. These findings have improved diagnosis, treatment, and prognosis. PMID- 9742308 TI - Growth disorders caused by genetic defects in the growth hormone pathway. AB - The growth hormone (GH) pathway is composed of a series of interdependent genes whose products are required for normal growth (Fig 1). The GH pathway genes include ligands (GH and insulin-like growth factor 1 [lGF-1]), transcription factors (prophet of pit 1, or prop 1 and pit 1), agonists and antagonists (growth hormone-releasing hormone [GHRH] and somatostatin), and receptors (GHRH receptor [GHRHR] and the GH receptor [GHR]). These genes are expressed in different organs and tissues, including the hypothalamus, pituitary, liver, and bone. Effective and regulated expression of the growth hormone pathway is essential for growth in stature as well as homeostasis of carbohydrate, protein, and fat metabolism. PMID- 9742309 TI - Investigating stem cells in the lung. AB - Disruption of the lung architecture by genetic events, environmental insults, or transformation can lead to respiratory diseases such as acute respiratory distress syndrome and lung cancer. Identification of the stem cells of the lung and the processes by which they regulate homeostasis may lead to better targets for treatment of these diseases. There are a number of approaches to study stem cell biology. Development of the lung and the major pulmonary cells of the bronchioles and alveolar regions of the lung is discussed in this review. Likewise, identifying the proteins that are critical for cell-specific expression and differentiation may identify approaches for manipulation of gene expression for use as therapy or treatment of lung diseases. Furthermore, strategies for studying stem cells in the lung are addressed by using the mouse as a model system. Gaining a more detailed understanding of the stem cells of the lung may provide new insight into the processes that govern lung biology and may lead to better treatments for lung diseases. Enthusiasm for the use of exogenous stem cells to replace tissue, organs, or other defective or deficient cells is boundless. Before stem cells can be used indiscriminately for these purposes, understanding tissue genetics and immunology is essential. Progress has been made in these areas for pulmonary disease. Attention to these models will be applicable to other organs and diseases. PMID- 9742311 TI - Psychodynamic psychotherapy, religious beliefs, and self-disclosure. AB - The intersection of psychodynamic psychotherapy and religious beliefs may present technical challenges for the psychotherapists; particularly if patients request to know more about the therapist's religious beliefs. Contrary to a recent technical recommendation for therapists to self-disclose personal religious beliefs when asked to do so, I suggest that such a request is complex and requires a thoughtful grounding in psychotherapeutic theory. Disclosing personal beliefs to patients runs the risk of being off-task as well as holding oneself out as an exemplar for the patient. Rather than adopt a formulaic response to requests for information, to deepen the understanding of the patient and the work of therapy, the therapist needs a complex understanding based on a careful diagnostic assessment of the patient, as well as an assessment of the current status of the psychotherapeutic venture. The workings of patients' particular transferences are often evident in requests for personal information and require careful evaluation and consideration. Likewise, countertransference elements may influence the type of response offered by the therapist. Using ethical principles as a guide is different from using them as a rule. The nexus of religious belief, psychosocial context, psychotherapy, and self-disclosure provides a potentially rich source of understanding when explored in the psychotherapeutic situation. PMID- 9742310 TI - The relationship between psychotherapist and prescribing psychiatrist. Some considerations. AB - We have reviewed the vagaries and vicissitudes of an increasingly common arrangement in contemporary mental health treatment, namely, the collaboration between psychotherapist and prescribing psychiatrist in the treatment of the patient who requires both psychotherapy and pharmacotherapy. Such a collaboration raises practical, legal, transference, counter-transference, economic, and training issues. It is our hope that this review of these issues will lead to further discussion as, in our experience, they have until now received scant attention in the mental health literature, in the day-to-day work of practitioners, and in supervision and training. We believe that successful collaboration can be achieved when sufficient awareness and planning are present at the outset. PMID- 9742312 TI - Characteristics of optimal clinical case formulations. The linchpin concept. AB - Being able to discern the presence of a central organizing linchpin in a given clinical case represents a highly advantageous state of affairs. One can, by virtue of this, proceed in a very efficient and economical, as opposed to piecemeal, fashion. Further, one can achieve this economy and efficiency without paying the price of superficiality, since one is getting to what might be termed "the heart of the matter" in the client's case. Finally, one has in a linchpin formulation a central blueprint that provides (a) a clear, constant goal for therapist and client; (b) a clarification for clients of both their power and of where and how they would best target their efforts; and (c) a vast heuristic suggestiveness as to how one might proceed therapeutically to bring about important change. PMID- 9742313 TI - Countertransference and life-and-death issues in group psychotherapy with child Holocaust survivors. AB - People, now in their fifties and sixties, who were children during the Nazi Holocaust in WWII, endured persecution, massive traumatization, the constant risk of being killed, as well as the violent loss of (most of) their family members. They have internalized the resulting ongoing confusion and conflicts as to whether they should be alive or dead. This is maintained as an integral part of the child component of their compound personality, described in this paper. During the three years of the psychotherapy group, on which we focused here, these issues were expressed in different ways, such as suicide threats, occasional intolerance to physically remaining in the group, and outbursts of annihilating rage at the therapists. The confusion and conflicts about the legitimacy and risks of their survival came to a head during the termination process we insisted upon. Much attention has also been given to the intricacies of our countertransference--further complicated by our own connection to the Holocaust. We learned, and described, just how essential it is to acknowledge and process this countertransference in order to both contain the intense affects of anxiety, rage, and mourning in the groups, and enable them to be safely expressed. We imagine similar dynamics can be expected in group therapy with other populations who suffered massive, man-made traumatization. PMID- 9742314 TI - Child abuse and neglect. Influences of qualitative research and clinical practice on child-care legislation and policy. AB - In an early paper, the authors presented the findings of a qualitative research study which applied the self-psychological and object relations theories to the social and interpersonal dynamics surrounding adolescent sexual offending. One of the findings of the study was that informal and formal social responses to detected offenders encouraged the rapid foreclosure of deviant, bad and dangerous social, interpersonal and sexual identity and therefore militated against therapeutic personality reconstruction. The current paper widens the scope of such observations to include the victim, as well as the offender, and examines the role of the therapist in mediating between intrapsychic and interpersonal priorities carried within the offense dynamics and socially and legally defined exigencies surrounding child abuse. The authors suggest that appropriate devolution of therapeutic agency can be devolved to patients through the concept of the twinship transference while at the same time attending to necessary psychiatric, medical, social and legal processes. PMID- 9742316 TI - Narrative lessons for the psychotherapist. Kafka's The Metamorphosis. AB - Literature has much to offer the psychotherapist. This paper has discussed some lessons for the psychotherapist contained in Franz Kafka's short story, The Metamorphosis. The therapist, like the therapist-reader of this story, can empathize with Gregor's monstrous change but still must hold him personally accountable. At the same time, the therapist-reader becomes increasingly impressed with the malignant nature of the Samsa household, and its role in generating Gregor's capacity for self-deception. The story also instructs about the paradox of catastrophe: Gregor is treated no less respectfully after his metamorphosis than he was before it. The therapist is thereby reminded of the centrality of feelings in human affairs. The constriction of Gregor's space does not cut him off from human feeling; rather, Gregor's inability to access, know, and take responsibility for his own feelings, especially his destructive ones, results in his constrictedness and detachment. In thinking about the story as dream, or in imagining a patient's account of a reality situation as if it were a dream, unseen mental process and content become more apparent. The disgusting, loathsome arrangements that people make with each other can evoke, be it in the therapist-reader or the therapist, reactions of aversion or hate. Such arrangements become more understandable when the importance, sometimes the necessity, of human attachment is appreciated. And finally, Kafka's The Metamorphosis alerts us to a sometimes but powerful preference and countertransference pitfall: we don't want to be bugged. PMID- 9742315 TI - Psycotherapy, managed care, and the economy of interaction. AB - This paper examines the disclosure over the value of long-term psychotherapy in a managed-care system. Many managed-care companies define extended psychotherapy as superfluous. Those who defend psychotherapy respond that the restrictions imposed by managed care are misguided and potentially harmful. After briefly discussing the relevant literature, the points of contention between psychotherapy and managed care are examined from the perspective of narrative literary theory. The analysis highlights the contrasting narrative assumptions implied about the importance of the clinical interaction. Pointing out each side's use of point-of view, narrative structure, and informational exchange, it is posited that beneath arguments that often focus on the commodities of time and money lie larger, conceptual differences. These stealthily serve to undermine the possibility of a rational debate. The paper concludes by asserting that psychotherapy and managed care assume incommensurate narratives of interaction when discussing the value of therapy. The terms of discourse must be expanded in order to account for the philosophical differences described. Several ways this might be accomplished are proposed. PMID- 9742317 TI - Severe childhood sexual abuse and nonverbal learning disability. AB - This article offers a review of, and case report on, the treatment of a young adult with a history of severe childhood abuse, dissociative symptoms, and right hemisphere dysfunction, or nonverbal learning disabilities (NLD). The core of nonverbal learning disabilities is the inability to synthesize information and create meaning from complex information. Learning is a form of adaptation and disruptions in an individual's meaning-making process. There are major implications for the person's overall adjustment. Trauma is itself complex and often damaging to the survivor's well being. Clinical assessment must take into account a person's cognitive style and possible learning deficits in order to adequately address traumatic material. Therapy must be modified in order to respond to the unique learning style of the NLD client. Finally, and important issue for therapists remains their willingness to broaden their awareness and knowledge base, and shift the treatment paradigm to meet the needs of the client with neurocognitive vulnerabilities. Treating clients with difficult trauma histories' alone, can elicit negative reactions in the therapist. Repeated experiences with a client's mistrust, anger, noncompliance or self-defeating habits are particularly stressful. The neuropsychological perspective can provide a valuable tool in the mastery of those reactions, and in building a context for empathy and a joint narrative. PMID- 9742318 TI - The case for evidence-based psychotherapy treatment guidelines. PMID- 9742319 TI - Aortic stenosis. Clinical evaluation and optimal timing of surgery. AB - Aortic valve disease is common in the elderly with recent data suggesting that aortic sclerosis and stenosis are the end-stage of an active disease process. Aortic atenosis may be diagnosed at symptom onset (angina, heart failure or syncope) but often the diagnosis is suspected in an asymptomatic patient with a systolic murmur. The diagnosis can be confirmed and disease severity evaluated reliably using Doppler echocardiography. Symptomatic severe aortic stenosis is treated with valve replacement, even in the elderly, due to the extremely poor prognosis without relief of outflow obstruction. Management is controversial when there is coexisting moderate aortic stenosis and left ventricular systolic dysfunction. PMID- 9742320 TI - Clinical assessment and management of mitral stenosis. AB - There have been significant advances in the diagnosis and treatment of the patient with mitral stenosis over the past two decades. Two-dimensional and Doppler echocardiography have supplanted the cardiac catheterization laboratory in the diagnosis and determination of the hemodynamic severity of the stenotic mitral valve. The development of a catheter-based approach for splitting fused commissures has led to earlier indications for intervention. It is likely that with the resurgence of rheumatic fever as well as influx of immigrant populations, the incidence of mitral stenosis may increase in the twenty-first century. It is thus important for the clinician to have a complete understanding of the evaluation and treatment options for the patient with mitral stenosis in the modern-day era. PMID- 9742321 TI - Assessment of valvular regurgitation with Doppler echocardiography. AB - Echocardiography is routinely performed for the evaluation of valvular regurgitation. Different applications of Doppler echocardiography have been successfully applied to detect and quantify valvular regurgitation. Recent advances in color Doppler made possible the study of the dynamic behavior of the regurgitant orifice and, along with continuous wave Doppler, can provide data on the regurgitant volume and fraction. Doppler echocardiography can also be used to follow serial changes in these hemodynamically important parameters after medical or surgical therapy. PMID- 9742322 TI - Management of mitral regurgitation. Optimal timing for surgery. AB - Chronic mitral regurgitation is a progressive disorder that can produce myocardial dysfunction in the absence of symptoms. Improvements in surgical techniques have resulted in earlier intervention, at times in asymptomatic patients. This article discusses the factors that influence prognosis, reviews the evidence supporting earlier intervention and provides guidelines for the management of patients with this lesion. PMID- 9742323 TI - Mitral valve repair vs replacement. Current recommendations and long-term results. AB - Techniques now exist to correct abnormalities of all components of the mitral valvular apparatus except extensive loss of pliable leaflet area. Thus, paradoxically, myxomatous valves with redundant leaflets represent the ideal candidates for mitral valve repair. Repair for mitral insufficiency can be performed for some rheumatic valves, but patient selection is critical. Loss of leaflet area, leaflet thickening, and extensive calcification of the leaflets or commissures are contraindications to repair. The abnormalities of the subvalvular apparatus are less important because a complete set of new chordae can be reconstructed using PTFE suture material. Some cases of endocarditis are ideal for repair using localized debridement and pericardial patch repair with or without PTFE chordal replacement. True ischemic mitral regurgitation of the Carpentier type I category is still something of a surgical enigma. Because it is a restrictive leaflet motion problem, annuloplasty alone is not always effective, and the outcome of any given repair attempt is less predictable. Repairs in patients with small annuli and multiple leaflet defects requiring complex series of maneuvers have a low probability of success. Furthermore, such patients with small left ventricular cavities are more prone to experience SAM. Several factors contributing to which therapy is chosen for mitral valve disease are summarized in Table 1. Patient selection, accurate evaluation of the cause or causes of mitral regurgitation, and well-executed application of the appropriate techniques for repair are all critical factors in the early and late success of mitral valve repair. PMID- 9742324 TI - Chronic aortic regurgitation. Role of medical therapy and optimal timing for surgery. AB - Aortic valve replacement should be performed once significant symptoms develop. Lacking important symptoms, operation should also be performed in patients with aortic regurgitation who manifest consistent and reproducible evidence of either LV contractile dysfunction at rest or extreme LV dilation. Noninvasive imaging techniques should play a major role in this evaluation. An important clinical decision, such as recommending aortic valve replacement in the asymptomatic patient, should not be based on a single echocardiographic or radionuclide angiographic measurement alone. When these data consistently indicate impaired contractile function at rest or extreme LV dilation on repeat measurements, however, operation is indicated in the asymptomatic patient. This strategy should reduce the likelihood of irreversible LV dysfunction in these patients and enhance long-term postoperative survival. PMID- 9742325 TI - Management of the patient with aortic root disease and aortic insufficiency. AB - There is no single standardized method of repair for the anatomic variations in aortic root pathology, which may include dissection, aneurysmal dilation, and valve disease and can occur at the annulus, sinuses of Valsalva, or the sinotubular junction. Composite valve/graft replacement, valve resuspension, and allograft each play a significant role in aortic root therapy, but none is applicable in all cases. Patient age, Marfan's syndrome, endocarditis, and previous valve replacement are examples of some of the wide variations in delineating factors. PMID- 9742326 TI - The role of intraoperative echocardiography in valve surgery. AB - The widespread use and popularity of intraoperative echocardiography (IOE) has resulted from advances in cardiac surgery, reparative procedures for valvular heart disease and, most specifically, mitral valve repair. IOE has grown exponentially and is becoming an integral part of the planning and evaluation of many types of surgical procedures such that it is now considered standard of care especially for the perioperative management of patients undergoing mitral and aortic valve repair. This article discusses the application of intraoperative echocardiography and focus specifically on valvular heart disease as this represents the most widely accepted indication for the procedure in current clinical practice. PMID- 9742327 TI - Choosing a prosthetic heart valve. AB - Although most of the available prosthetic heart valves function remarkably well, the variety of available choices attests to the inability of any single one to fulfill the requirements of the ideal valve substitute. The mechanical prostheses include the caged-ball, tilting-disc, and bileaflet valves. Tissue valves available in the United States are the Carpentier-Edwards and Hancock porcine heterograft valves and the Carpentier-Edwards pericardial valve. Review of several large comparative studies on valve performance reveals that the overall results with tissue and mechanical valves are about equal at the end of 10 years. The characteristics of each type of valve substitute dictate the selection of one prosthesis in preference to others for a particular patient. Mechanical prostheses are recommended for patients without contraindications for anticoagulants. Tissue valves are reserved for patients over 65 years of age or for patients in whom anticoagulation is contraindicated. Multiple other patient related factors need to be considered in selecting the appropriate valve, including the psychosocial situation and patient preference. PMID- 9742328 TI - Evaluation of prosthetic valve function and associated complications. AB - Significant advances in imaging modalities have occurred to evaluate prosthetic valve function and associated complications. These developments involve predominantly the introduction of Doppler technology for the non-invasive determination of gradients and valve areas and TEE for an improved assessment of valve structure, function, and associated complications. The current role of cinefluoroscopy is mostly to complement TEE in the evaluation of motion of mechanical prosthetic valves in the aortic position. Cardiac catheterization is now rarely needed to assess valve function. Diagnosis of prosthetic valve obstruction can be performed in the majority of cases with transthoracic Doppler echocardiography. Differentiation of valve obstruction from normal valve function in small valves with high flow conditions, however, may be difficult. Because of this and the variability in normal valves among different prostheses, knowledge of the type and size of the implanted valve is essential. Patients and ultrasound laboratories are encouraged to seek and provide this information on a routine basis. Although transthoracic echocardiography is the main diagnostic modality for the serial evaluation of prosthetic valve function, it is important to recognize its limitations in assessing prosthetic mitral regurgitation and evaluating structural abnormalities of prosthetic valves. These are the situations in which TEE has the most impact. A summary of general indications of TEE in prosthetic valves is provided in Table 6. Finally, a baseline transthoracic Doppler study is essential in the overall follow-up and serial evaluation of valve function. For future comparisons, the best indices of valve functions are those obtained for patients as their own control, from a baseline Doppler echocardiographic study performed early after the operation. PMID- 9742329 TI - Valvular disease associated with systemic illness. AB - The connective tissue diseases are immune-mediated inflammatory diseases that manifest predominantly with symptoms and signs of musculoskeletal and mucocutaneous inflammation. They frequently affect the heart valves, pericardium, and myocardium. In patients with AKS, the aortic root and conduction system are also frequently involved. Echocardiographic series in these patients have demonstrated that valvular disease is highly prevalent and associated with substantial morbidity and mortality (Table 1). The prevalence rates of clinically detected valvular disease, however, are either unknown or low. This discrepancy is related to lack of awareness, overshadowing of the cardiovascular manifestations by the inflammatory symptoms and signs of the musculoskeletal system, lack of systematic application of the history and cardiovascular physical examination, and high sensitivity of echocardiography for detecting subclinical abnormalities. Several valvular abnormalities have been identified as unique to a specific disease. Libman-Sacks vegetations, valve nodules, and subaortic bump are characteristic of SLE, RA, and AKS (see Table 1). The valvular complications and respective therapy are similar to those of other causes of valvular disease; however, the associated morbidity and mortality of these complications in these patients are high. The worse prognosis of valvular disease in these patients is related to the chronicity and debilitating nature of their illness, their high prevalence of multisystem disease, and immunosuppression. These factors underscore the importance of early recognition, prevention of complications, and proper clinical or echocardiographic follow-up. The distinctive echocardiographic characteristics of the valve abnormalities associated with the connective tissue diseases may allow their differentiation from other common valvulopathies, such as infective endocarditis, rheumatic valvular disease, and degenerative valvular disease (Table 2). Despite the clinical and prognostic implications of valvular disease associated with the connective tissue diseases, incomplete data are available about pathogenesis, relation to clinical features of the primary disease, evolution, and effect of steroid or cytotoxic therapy. Echocardiography, especially TEE, has the potential to redefine the prevalence rates and to characterize better the valve abnormalities associated with these conditions. Finally, future large cross-sectional and longitudinal studies using clinical and echocardiographic data may help to define better the presence, evolution, and therapy of the valvular disease associated with the connective tissue diseases. PMID- 9742330 TI - Tricuspid valve disease. Clinical evaluation, physiopathology, and management. AB - The tricuspid and mitral valves are homologous whose function depends on coordination among components. Isolated tricuspid valve abnormalities are relatively uncommon. Rheumatic disease, chemicals, immunologic and degenerative disorders alter leaflet anatomy and may result in either stenosis, insufficiency or a combination. More often, tricuspid disorders present as a component of congenital syndromes or secondary to pulmonary vascular or let heart disease which alter geometry and function of nonleaflet components. PMID- 9742331 TI - Valvular heart disease in pregnancy. A contemporary perspective. AB - Valvular heart disease may have a significant impact on the course and outcome of pregnancy with implications for fetal as well as maternal health. Optimally, serious symptomatic valvular heart disease should be detected and treated before pregnancy. Whether a pregnant woman is known to have valvular heart disease or is diagnosed during pregnancy, it is imperative that she is managed by an experienced multidisciplinary team. Although medical therapy may alleviate symptoms of heart failure in some patients, definitive intervention either with percutaneous balloon valvuloplasty or with surgical valve replacement may be necessary. PMID- 9742332 TI - Cardiac arrhythmias. AB - Supraventricular dysrhythmias (SVDs) present the most frequent medical complication after thoracic surgery and have been associated with prolonged hospital stays. The reported incidence of SVDs in the thoracic surgery patient population ranges from 10% to 40%, with factors such as age and extent of surgery markedly influencing the incidence. This article focuses on new issues leading to improved understanding of the pathophysiology and mechanisms of SVDs after surgery. New approaches directed at prophylaxis and acute therapy of SVDs are discussed, as are recommendations to prevent thromboembolic events due to atrial dysrhythmias following thoracic surgery. PMID- 9742333 TI - Myocardial ischemia and infarction postthoracotomy. AB - The best long-term survival for any given lung cancer patient is provided by surgical resection. However, pneumonectomy still has the highest mortality rates, often due to cardiac complications. Risk assessment can be aided by preoperative evaluation of thoracic surgery patients. The role of right heart function, intraoperative management, and postoperative conditions in myocardial ischemia and infraction are analyzed, and the benefits of different kinds of resection are weighed in light of possible cardiac complications. PMID- 9742334 TI - Postoperative atelectasis. AB - Postoperative atelectasis is a common problem following any surgery. Limited atelectasis is usually well-tolerated and easily reversible. However, complete atelectasis of the remaining lung following partial lung resection may be poorly tolerated. Thoracic surgical procedures increase the risk because pain, thoracic muscle injury, chest wall instability, and diaphragmatic dysfunction impair clearance of secretions by cough. In addition, patients with lung diseases are prone to increased bronchial secretions. Prophylaxis includes preoperative and postoperative physiotherapy and medications, which should be graded in accordance to the individual patient's risk factors. Large atelectasis requires bronchoscopy to remove mucous plugs. Tracheostomy should be considered in patients with relapsing atelectasis or swallow disorders. PMID- 9742335 TI - Postoperative pneumonia. AB - Thoracic surgical patients are susceptible to pneumonia because of impaired systemic and lung host defenses. The incidence of pneumonia is higher with more extensive lung resections. Current prophylactic antibiotic therapy is based primarily on general surgical experience with emphasis on wound infection, not pneumonia. With expansion of indications for lung resection to include higher risk patients, there is a need to render antibiotic prophylaxis more specific to bacteria causative of pneumonia. PMID- 9742336 TI - Preparation of high-risk patients for major thoracic surgery. AB - For a least a decade, many patients have benefitted from new indications of major thoracic surgery owing to improvements in the surgical and anesthetic procedures of thoracic surgery. Identification of risk factors of perioperative morbidity and mortality becomes of paramount importance when trying to lesson the postoperative mortality rate to 1% or less. The careful assessment of the candidates for thoracic surgery with a multidisciplinary approach is the cornerstone of such an objective. The lower mortality rate should be achievable with a preoperative preparation of the patients of a rehabilitation and nutritional program and a pharmacologic treatment optimization. PMID- 9742337 TI - Adult respiratory failure. AB - Pulmonary complications following thoracic surgery are common and associated with significant morbidity and mortality. In particular, acute respiratory distress syndrome (ARDS) can occur postoperatively or after trauma. This syndrome, when complicated by multisystem organ failure, often leads to a poor outcome. This article describes the etiology and pathophysiology of ARDS and evaluates recent advances in pharmacological and nonpharmacological therapies. In addition, newer modalities of mechanical ventilatory support are reviewed. PMID- 9742338 TI - Mechanical ventilation for respiratory failure postthoracotomy. AB - Although the exact incidence of postthoracotomy respiratory failure is unknown, it can be estimated to lie between 5% and 15%, given that many of these patients suffer from comorbid cardiopulmonary disease. Preoperative assessment is essential to more accurately predict those patients at increased risk for the development of respiratory failure. Efforts to minimize these risks include the aggressive use of analgesics. In the event of respiratory failure, the clinician must have a clear understanding of the underlying cardiopulmonary pathology, if any, and of the impact of the anesthetic and surgical procedures on cardiopulmonary performance. The twin goals of mechanical ventilation should be to provide inspiratory muscle assistance and rest while preventing the onset of muscle atrophy. To that extent, the limitations of the various modes of mechanical ventilation must be appreciated as to their impact on patient ventilatory performance. Weaning, which should be regarded simply as an extension of mechanical ventilation, should be approached with an appreciation of the pathophysiologic basis underlying ventilatory failure, the factors responsible, and a rational approach to their repair. PMID- 9742339 TI - Postpneumonectomy pulmonary edema. AB - The adult respiratory distress syndrome seen after pneumonectomy is an uncommon but usually lethal complication. Its etiology remains unknown, although several factors such as fluid overload, endothelial damage, lymphatic interruption, and hyperinflation are thought to be involved in its pathogenesis. PMID- 9742340 TI - Neurologic complications in thoracic surgery. AB - In this article, a significant number of neurologic conditions have been presented that have importance to the thoracic surgeon. The most important point, however, is that most of the neurologic complications are avoidable by careful surgical technique and preoperative and postoperative care. PMID- 9742341 TI - Gastrointestinal complications postthoracotomy and postvagotomy. AB - Postthoracotomy gastrointestinal complications, although relatively uncommon, can be associated with significant morbidity and mortality. It is necessary to identify patients who are at high risk for gastrointestinal complications during the preoperative evaluation. Appropriate stress ulcer prophylaxis should be provided to high-risk patients, and enteral feeds should be initiated as early in the postoperative course as possible. Postoperative hypotension and massive blood transfusions can be avoided with early reexploration in the case of postoperative hemorrhage. Finally, unexplained abdominal pain must not be ignored; a high index of suspicion should be maintained, with early and liberal use of diagnostic tools such as standard radiography, CT, endoscopy, and angiography. Consultation should be requested from a surgeon experienced in abdominal catastrophes. Early laparotomy with aggressive operative management can be lifesaving therapy but must be not applied in a cavalier fashion, as many of these disorders can and should be managed conservatively. PMID- 9742342 TI - Complications of multimodality therapy. AB - Multimodality therapy may further increase treatment-related complications. However, such therapy may be necessary for advanced or biologically aggressive tumors, and information gained from patient entry into prospective trials may attenuate current and future treatment-related complications. Ad hoc use of multimodality therapies is generally not recommended. PMID- 9742343 TI - Adverse effects of medications commonly administered to thoracic surgical patients. AB - A wide variety of medications is commonly used following thoracic surgical procedures. All of these medications have associated side effects that may adversely affect the recovery of patients. A complete understanding of the important adverse effects of all the medications used postoperatively can limit or eliminate unwanted medication effects and lead to a more successful outcome. This article will review the important actions and side effects of the most commonly administered medications following thoracic surgical procedures. PMID- 9742344 TI - Pathogenesis and management of persistent postthoracotomy pain. AB - Persistent chest wall pain is common after thoracotomy and is usually caused by recurrence or progression of malignancy. It should prompt efforts to identify and treat the causative disease. A minority of patients experience persistent pain not related to neoplasm. This pain may last for years, but is usually not severe. A small subset of these patients experience persistent severe pain, which may be debilitating. The pain may be owing to various causes. Diagnosis and treatment should be individualized and directed toward the causes believed to be present. First-line pharmacologic therapies include NSAIDs, tricyclic antidepressants, antiepileptics, and low-dose opioids. Some patients require more sophisticated treatment from multidisciplinary pain-management clinics. This treatment may include nerve blocks, physical therapy, sympathectomy, cryoneurolysis, or long term neuromodulation with epidural analgesia or spinal cord stimulation. Because of the severe pain these patients may experience and the difficulty and expense associated with treatment, prevention may be the best strategy for dealing with this problem. Recent laboratory and clinical studies indicate that minimizing perioperative pain can suppress certain alterations in the nervous system that may prevent the genesis and maintenance of chronically painful conditions. This suggests that strategies for avoiding PTPS may begin with aggressive perioperative anesthetic and analgesic techniques. More effective application of knowledge already available from laboratory studies awaits further clinical trials. New drugs such as NMDA inhibitors hold promise for more effective treatment in the future. PMID- 9742346 TI - Ambulatory obstetric management. PMID- 9742345 TI - The dying thoracic patient. AB - Health care providers should understand that the practice of good medicine includes not only diagnosing and curing diseases, but also effectively communicating with patients and families and helping terminally ill patients die a peaceful and dignified death. Patients in America come from varied backgrounds, and it is important for physicians to consider cultural and religious issues. Physicians should combine their clinical judgment with objective outcome data to provide optimal care for patients. Informed consent should be obtained from patients after offering a detailed plan of care that would include appropriate interventions and the consequences of no intervention. The physician should then assist the patient in making a decision that would provide the best possible future for that individual. The four fundamental principles of biomedical ethics, namely beneficence, nonmaleficence, autonomy, and justice, should be considered when analyzing an ethical problem. Voluntary active euthanasia, which means performing a deliberate act (e.g., administering a lethal injection) to end a patient's life, should not be performed by a physician. Withholding and withdrawing basic and advanced life support constitutes passive euthanasia. Good communication with patients early in the clinical course whenever possible results in an ethically correct decision. A nonconfrontational, sympathetic, and compassionate approach to family members and legal surrogates facing the immediate death of their loved ones leads to the best possible outcome. It is the duty of the physician to assure the patient and the family that he or she will not abandon the patient. Effective communication is the key to solving almost all ethical dilemmas when caring for the dying thoracic patient. PMID- 9742347 TI - Ambulatory management of preterm labor. AB - The care of women with preterm labor has focused predominantly on inpatient therapy: tocolysis, antibiotics, and steroid administration. The emphasis is slowly but surely shifting to secondary prevention and outpatient therapy. Our goal should be toward primary prevention of preterm labor in all women. Then and only then will a true reduction in spontaneous prematurity rates be seen. PMID- 9742348 TI - The home management of preterm premature ruptured membranes. PMID- 9742349 TI - Pyelonephritis. PMID- 9742350 TI - Placenta previa, placenta abruptio. PMID- 9742351 TI - Hypertension. PMID- 9742352 TI - Anticoagulation. AB - Outpatient management of women requiring treatment and prophylaxis against thromboembolic conditions during pregnancy and the postpartum period requires a coordinated effort between the patient, her obstetrician and, in certain cases, a hematologic consultant. The anticoagulation regimen should be tailored to the clinical situation, with patient compliance and cost taken into consideration. PMID- 9742353 TI - Outpatient management of asthma during pregnancy. PMID- 9742354 TI - Ambulatory management of multiple gestation. AB - Ambulatory management of multiple gestation requires careful and continuing care by the obstetrician. The initial evaluation should include a comprehensive history, including use of fertility enhancing drugs and ART, family history, social history; a general physical examination, including a pelvic examination; laboratory evaluation, including complete blood cell count, dipstick urinalysis for protein and glucose, urine culture, blood type, Rh factor and irregular blood antibody determination, serology for rubella, syphilis, hepatitis B surface antigen and varicella (if there is no history). A Papanicolaou smear should be done at the time of the pelvic examination, as should evaluation for bacterial vaginosis. Ultrasound assessment of placentation should be done at 14 weeks' gestation, but vaginal or perineal ultrasound of cervical length should be done at the initial visit. Other testing procedures should include repeat ultrasound evaluation for fetal growth every 4 weeks in a dichorionic placentation and every 3 weeks if monochorionic placentation is present. Triple screen MSAFP at 16-18 weeks' gestation and blood sugar screening at 22-26 weeks should be performed. After the first trimester, the patient should schedule physician visits every 2 weeks or less. Routine medications should include one prenatal vitamin per day, additional folic acid supplementation of 1.0 mg per fetus, supplemental iron preparation, and additional calcium to equal 1500 mg/day. The use of low-dose aspirin to prevent preeclampsia in twin gestations has not been adequately studied. Continuing vigilance by the knowledgeable obstetrician should occur. Multiple gestations should not be cared for by non-physician providers or by family physicians. Referral to a maternal-fetal medicine unit is recommended. PMID- 9742355 TI - Ambulatory care of the pregnant woman with diabetes. PMID- 9742357 TI - Cervical ripening. AB - In conclusion, no pharmacologic method presently used in the hospital to ripen the cervix has attained the level of standard of care for application in most office or clinic outpatient settings. Published data are accumulating that support the safety of the prostaglandin preparations for this purpose. Sweeping the membranes in the office and clinic does have sufficient data to support its continued use for this purpose. PMID- 9742356 TI - Hyperemesis gravidarum. PMID- 9742358 TI - Early postpartum discharge. PMID- 9742359 TI - Psychosocial impact of high-risk pregnancy: hospital and home care. AB - This article has reviewed research on the psychosocial impact of high-risk pregnancy on women undergoing antepartum hospitalization or home care, and discussed implications for health care professionals arising from the results of these studies. It is evident that high-risk pregnancy is very stressful for pregnant women and their families, irrespective of the setting of care. However, hospitalization seems to be associated with a greater number of stressors, the most predominant being separation from home and family. Wherever possible, health care professionals should strive to develop safe and acceptable alternatives to antepartum hospitalization. PMID- 9742361 TI - Antepartum testing. AB - Among the methods of antepartum testing in use today, the nonprovocative tests (NST, BPP, MBPP) are safe and effective for use in ambulatory settings. Outpatient or office utilization of the CST is limited by the need for intravenous access (when oxytocin is used) and by the potential for uterine hyperstimulation and resultant acute FHR abnormalities. Regardless of the method of testing used, large studies have confirmed that the fetal death rate among patients undergoing antepartum testing is significantly lower than that in the general, untested population. This is a particularly encouraging observation in view of the fact that antepartum testing is used almost exclusively in complicated pregnancies at highest risk for poor outcome. In the future, protocols using adjunctive testing methods (fetal movement counting, fetal movement profile, Doppler velocimetry) in combination with standard methods (CST, NST, BPP, MBPP) may further reduce the incidence of fetal death in high-risk populations. At present, the beneficial effects of antepartum testing have created a situation in which the likelihood of fetal death in high-risk, tested populations is lower than that in low-risk, untested populations. This paradox will force us to consider the option of routine antepartum testing in all pregnancies. PMID- 9742362 TI - Initial assessment: the history in women with pelvic floor problems. AB - History-taking from women with pelvic floor problems becomes easier with practice. In the time it has taken you to read this chapter, the histories of several patients could have been ascertained. Efficiency during the focused history will assume increasing importance as time constraints tighten. Combined use of written and verbal assessments provides maximum information in a concise fashion. This technique allows the treating physician to educate the patient while being sensitive to her individual concerns, such as her specific preferences for treatment and her social/lifestyle considerations. Efficient medical and/or surgical recommendations can be given within this context. These recommendations can reflect the preferred treatment given the milieu of the entire pelvic floor anatomy and function after physical examination and any appropriate testing. PMID- 9742360 TI - Ambulatory obstetrical care: strategies to reduce telephone liability. AB - The telephone will become the centerpiece of ambulatory care services. As such, a pertinent aspect of office procedures will necessarily include a protocol to manage and document telephone calls. Encourage your office staff to use good telephone manners, as listed in Table 5. The net result should be a reduction in telephone liability risks and an enhanced reputation for your office. PMID- 9742363 TI - Physical examination and pretreatment testing of the incontinent woman. PMID- 9742364 TI - Urodynamic evaluation for female urinary incontinence. PMID- 9742365 TI - Pharmacologic therapy for urinary incontinence and voiding dysfunctions. PMID- 9742366 TI - Nonsurgical therapies for urinary incontinence. AB - Many nonsurgical treatments for incontinence are available and all offer benefit to some patients. Patients with mild to moderate incontinence usually benefit the most from these modalities and, for many, it may obviate the need for surgery. Several factors influence the success of nonsurgical treatments for incontinence. Most important is the patient's motivation and ability to comply with the therapy. Selection of therapy appropriate for a given patient is, therefore, important. Compliance is better when the patient has a better understanding of the therapy and expected results. When appropriate, combinations of therapy may be very useful. PMID- 9742367 TI - Common operations for stress incontinence: selecting the correct operation. PMID- 9742368 TI - Why anti-incontinence surgery succeeds or fails. PMID- 9742369 TI - Postoperative urinary drainage. PMID- 9742370 TI - Urinary tract infections. AB - Urinary tract infection in women has its origin, predominantly, via ascending bacteria from the periurethral microflora. Asymptomatic bacteriuria, except for the pregnant patient, need not be treated. E. coli is the most common bacterium to cause UTIs, and is usually susceptible to oral antibiotics. Patients who are hospitalized with an indwelling Foley catheter or who have undergone instrumentation, tend to be infected with a bacterium other than E. coli. Patients with uncomplicated cystitis can effectively be treated with an oral antibiotic (Table 1) for 3 days. Patients who do not respond to empiric therapy have a recurrence within 2 weeks of treatment, or who have a recurrence within the first week after treatment, should have a pretreatment. PMID- 9742371 TI - Bladder and ureteral injury: prevention and management. PMID- 9742372 TI - Cystourethroscopy for the practicing gynecologist. PMID- 9742373 TI - Urinary incontinence and concomitant prolapse. PMID- 9742374 TI - Surgical management of pelvic organ prolapse and stress urinary incontinence. AB - There is no single operative approach to correct pelvic organ prolapse in conjunction with urinary and/or fecal incontinence or rectal prolapse. Each case needs to be individualized and dealt with surgically following the principles outlined in Table 1. In postmenopausal women, it is not only important to pretreat patients with estrogen prior to reconstructive pelvic surgery, but also maintain patients on long-term treatment after surgery. The genitourinary and reconstructive pelvic surgeon should have the skills to offer patients alternative approaches tailored to their individual symptomatology, and anatomic and pelvic pathology. Long-term follow-up of all patients is imperative to ascertain the clinical and cost effectiveness of these procedures. PMID- 9742375 TI - The surgical pathologist's approach to fine needle aspiration. AB - The diagnostic process begins with triage of the FNAB, depending on the patient's problem. Portions of the specimen can then be set aside for appropriate immunocytochemistry, hormone receptors analysis, electron microscopy, flow cytometry, or molecular studies. Microscopic evaluation, as in surgical pathology, begins with scanning objective examination of tissue architecture, followed by study of cellular characteristics, and finally the nuclear features. Cytopathology and surgical pathology are no longer distinct entities. PMID- 9742376 TI - Fine needle aspiration cytology of papillary neoplasms. AB - This article covers the fine needle aspiration biopsy cytomorphology of papillary carcinomas of different organs, differential diagnoses, and clinical correlation. Diagnostic problems and helpful cytologic features are emphasized. The purpose is to have a concise source of information that helps the pathologist to evaluate these neoplasms. PMID- 9742377 TI - Fine needle aspiration biopsy of the pancreas. AB - This article covers basic topics such as indications, contraindications, techniques and complications. Individual sections focus on the differential diagnosis between adenocarcinoma and benign or reactive processes, diagnosis of pancreatic endocrine tumors, and the diagnosis of cystic lesions using pancreatic cyst fluid analysis including cytopathology. PMID- 9742378 TI - Fine needle aspiration cytology of the kidney, renal pelvis, and adrenal. AB - Fine needle aspiration biopsy for cytologic evaluation of mass lesions of the kidney, renal pelvis and adrenal is a safe economic and accurate diagnostic procedure. Tumors and inflammatory or degenerative lesions arising from these organs have characteristic cytologic features permitting their correct identification in the majority of cases. PMID- 9742379 TI - Fine needle aspiration cytology of the thyroid. AB - This article is an up-to-date review of thyroid fine needle aspiration. Special emphasis is placed on pitfalls and differential diagnoses of common and uncommon thyroid lesions. PMID- 9742380 TI - Fine needle aspiration biopsy of the liver. Principal diagnostic challenges. AB - Fine needle aspiration biopsy (FNAB) is the diagnostic procedure of choice for evaluation of liver lesions. Although primarily applied to malignant disease, it is also used in the evaluation of benign conditions. Improvements in imaging techniques and advances in cytologic interpretations, as well as production of new biopsy needles, have all contributed to the rapid increase in radiologically guided liver FNAB. PMID- 9742381 TI - Fine needle aspiration of soft-tissue lesions. AB - The heterogeneity of soft-tissue neoplasms constantly challenges the diagnostic skills of pathologists. Fine needle aspiration biopsy (FNAB) cytology can provide a quick and relatively safe assessment of soft-tissue masses and can significantly contribute to patient management. This article illustrates the FNAB cytologic features of the most commonly encountered soft tissue lesions and discusses their differential diagnoses. PMID- 9742382 TI - Fine needle aspiration cytopathology of malignant lymphoma. AB - Morphologic features allowing the cytopathologist to distinguish a reactive lymph node from a malignant lymphoproliferative disorder and to distinguish Hodgkin lymphoma from non-Hodgkin lymphoma are presented in concert with pertinent immunophenotypic profiles of various lymphomas. The limitations and diagnostic pitfalls of aspiration cytopathology in the diagnosis of lymphoma and lymphoid aspirates are also discussed. PMID- 9742383 TI - If cells could talk. The application of new techniques to cytopathology. AB - Cytopathology is no longer simply a screening modality limited to the "Pap mills" of yore. The news in cervicovaginal cytology is automation. The news in FNA cytology is the application of molecular techniques. Whether it is the detection of specific proteins/antigens for definitive diagnoses/treatment guidance in immunotherapy, or it is "reading nucleic acids," the cytopathologist of the future will be called upon to gather and report more detailed and precise information. As we develop methods for extrapolating the secrets previously locked within the individual cells, it becomes evident that the cells were talking all along, we just did not know how to listen. PMID- 9742384 TI - Electronic imaging in cytopathology. AB - Electronic imaging is becoming important for conveying instructional and diagnostic information. This article presents an overview of the acquisition and display of microscopic images for day-to-day cytologic and histologic practice. The devices, software, and methods for image capture and manipulation are described. PMID- 9742385 TI - Medicolegal issues and fine needle aspiration of the breast. What you can do to decrease your risk of being sued. AB - Carcinoma of the breast is the most litigated cancer in women. This article discusses steps that can be taken to reduce the risk of being sued. Topics covered include laboratory practices, patient contact, informed consent, documentation, record keeping, interpretive problems, triple test, diligence, and reporting results as well as what to do if sued. PMID- 9742386 TI - Innate immunity. PMID- 9742387 TI - Retinal processing: amacrine cells keep it short and sweet. AB - A recent study suggests a neuronal circuit in the retina by which amacrine cells contribute to the generation of transient responses in ganglion cells, thereby enabling the visual system to detect changes in light intensity. PMID- 9742388 TI - Cholesterol homeostasis: a role for oxysterols. PMID- 9742389 TI - Antigen presentation: coming out gracefully. AB - Efficient assembly of antigen-presenting class I MHC molecules requires the formation of a complex between the class I molecule and the TAP peptide transporter. The complex has been found to contain an additional four proteins, which help to ensure optimal peptide loading onto the class I molecules. PMID- 9742390 TI - Transport vesicles: coats of many colours. AB - Clathrin-coated vesicles transport proteins and membranes between intracellular compartments. Adaptor molecules determine vesicle specificity; recently, a third type of adaptor protein, AP3, has been identified and implicated in the biogenesis of endosomal and lysosome-related organelles. PMID- 9742391 TI - Synaptic transmission: spillover at central synapses. AB - Recent studies suggest that transmitter molecules released at central synapses sometimes diffuse long enough distances to activate receptors located outside the synaptic cleft or even in neighboring synapses. This transmitter 'spillover' may have important physiological consequences. PMID- 9742392 TI - Transcription: activation by cooperating conformations. AB - The cooperative formation of a higher-order complex between transcription factors NFAT and Fos-Jun is accompanied by conformational changes in both DNA and protein. This allows formation of an extended interface between the proteins, while conserving recognition of the core DNA binding sequences. PMID- 9742393 TI - Organelle division: from coli to chloroplasts. AB - FtsZ, an ancestral homolog of eukaryotic tubulin, assembles into the cytokinetic Z ring that directs cell division in bacteria. Recent results indicate that FtsZ is also used for division by chloroplasts, though not by mitochondria. PMID- 9742394 TI - DNA repair: the Nijmegen breakage syndrome protein. AB - The gene mutated in Nijmegen breakage syndrome, a chromosome instability disorder, has been identified and sequenced. The protein product of this gene forms a complex with hMre11 and hRad50--proteins that are involved in repairing double-strand breaks in DNA. PMID- 9742395 TI - Byr4 and Cdc16 form a two-component GTPase-activating protein for the Spg1 GTPase that controls septation in fission yeast. AB - BACKGROUND: Spatial and temporal control of cytokinesis ensures the accurate transmission of genetic material and the correct development of multicellular organisms. An excellent model system in which to study cytokinesis is Schizosaccharomyces pombe because there are similarities between cytokinesis in S. pombe and mammals and because genes involved in S. pombe cytokinesis have been characterized. In particular, formation of the septum is positively regulated by the Spg1 GTPase and its effector, the Cdc7 kinase. Septation is negatively regulated by Cdc16, a protein similar to GTPase-activating proteins (GAPs) for Ypt GTPases, and by Byr4, a protein of unknown biochemical function. This study investigates the relationship between Byr4, Cdc16, and Spg1. RESULTS: Genetic interactions were observed between byr4, cdc16, and spg1 mutants. Byr4 bound to Cdc16 and Spg1 in yeast two-hybrid assays and in coprecipitations in vitro and in yeast. Byr4 inhibited the dissociation and hydrolysis of GTP bound to Spg1, but when Byr4 and Cdc16 were combined together they displayed Spg1GAP activity in vitro; Cdc16 alone had no detectable GAP activity. The binding of Byr4 to Spg1 and the Byr4-Cdc16 Spg1GAP activity were specific because Byr4 and Cdc16 did not bind to or affect the GTPase activities of the seven known S pombe Ypt family GTPase. CONCLUSIONS: Byr4 and Cdc16 form a two-component GAP for the Spg1 GTPase. Byr4 and Cdc16 appear to negatively regulate septation in S. pombe by modulating the nucleotide state of Spg1 possibly in a spatially or temporally controlled manner. PMID- 9742396 TI - Apparent caspase independence of programmed cell death in Dictyostelium. AB - During normal development, cell elimination [1,2] occurs by programmed cell death (PCD) [3], of which apoptosis [4] is the best known morphological type. Activation of cysteine proteases termed caspases [5] is required in many instances of animal PCD [6-9], but its role outside the animal kingdom is as yet unknown. PCD occurs during developmental stages in the slime mold Dictyostelium discoideum [10,11]. Under favorable conditions, Dictyostelium multiplies as a unicellular organism. Upon starvation, a pathway involving aggregation, differentiation and morphogenesis induces the formation of a multicellular fungus like structure called a sorocarp [12], consisting mainly of spores and stalk cells, the latter being a result of cell death. Dictyostelium cell death is similar to classical apoptosis in that some cytoplasmic and chromatin condensation occurs but differs from apoptosis because it involves massive vacuolisation and, interestingly, lacks DNA fragmentation [11]. We examined whether caspase activity is required for Dictyostelium cell death. We found that caspase inhibitors did not affect cell death, although some caspase inhibitors that did not inhibit cell death impaired other stages in development and could block affinity-labelling of soluble extracts of Dictyostelium cells with an activated caspase-specific reagent. The simplest interpretation of these results is that in Dictyostelium, whether or not caspase-like molecules exist and are required for some developmental steps, caspase activation is not required for cell death itself. PMID- 9742397 TI - The WASp homologue Las17p functions with the WIP homologue End5p/verprolin and is essential for endocytosis in yeast. AB - Several end mutations that block the internalisation step of endocytosis in Saccharomyces cerevisiae also affect the cortical actin cytoskeleton [1]. END5 encodes a proline-rich protein (End5p or verprolin) required for a polarised cortical actin cytoskeleton and endocytosis [2,3]. End5p interacts with actin [4], but its exact function is not yet known. To help elucidate End5p function, we sought other End5p-interacting proteins and identified the LAS17/BEE1 gene (encoding the yeast homologue of the human Wiskott-Aldrich Syndrome protein, WASp) as a high-copy-number suppressor of the temperature-sensitive growth and endocytic defects of end5-1 cells (carrying a frameshift mutation affecting the last 213 residues of End5p). LAS17 is unable to suppress a full deletion of END5 (end5 delta), however, suggesting that the defective End5-1p in end5-1 mutants may be stabilised by Las17p. The amino terminus of Las17p interacts with the carboxyl terminus of End5p in the yeast two-hybrid system and similar interactions have been shown between WASp and a mammalian End5p homologue, WASp interacting protein (WIP) [5]. As las17 delta deletion mutants are blocked in endocytosis, we conclude that Las17p and End5p interact and are essential for endocytosis. PMID- 9742398 TI - Conjugation in S. pombe: identification of a microtubule-organising centre, a requirement for microtubules and a role for Mad2. AB - During the G1 phase of the cell cycle, cells of the fission yeast Schizosaccharomyces pombe can be induced to mate by nitrogen starvation and the presence of mating pheromones. Polarised growth towards cells of the opposite mating type (P or M) leads to the formation of a projection tip and, upon contact, localised cell wall degradation results in conjugation and cell fusion [1]. Here, we have investigated the role of microtubules in this process. We describe a previously unidentified microtubule-organising centre (MTOC) that forms at projection tips upon cell-to-cell contact, before cells fuse. Treatment of mating cells with the microtubule-destabilising drug thiabendazole (TBZ) showed that microtubule integrity was required for mating at two distinct stages: during projection tip formation and cell fusion. Projection tip formation requires filamentous (F) actin function [2] and microtubules are required for the localisation of F actin to the projection tip. We also identify a role during mating for Mad2--a mitotic checkpoint protein that is required in all eukaryotes to maintain the mitotic state in response to microtubule depolymerisation [3]. S. pombe mad2 mutant cells were compromised in their ability to mate upon removal of TBZ, indicating that in fission yeast, in the absence of microtubules, Mad2 is also required to maintain mating competence. PMID- 9742399 TI - Actin cytoskeleton organization regulated by the PAK family of protein kinases. AB - Cdc42, Rac1 and other Rho-type GTPases regulate gene expression, cell proliferation and cytoskeletal architecture [1,2]. A challenge is to identify the effectors of Cdc42 and Rac1 that mediate these biological responses. Protein kinases of the p21-activated kinase (PAK) family bind activated Rac1 and Cdc42, and switch on mitogen-activated protein (MAP) kinase pathways; however, their roles in regulating actin cytoskeleton organization have not been clearly established [3-5]. Here, we show that mutants of the budding yeast Saccharomyces cerevisiae lacking the PAK homologs Ste20 and Cla4 exhibit actin cytoskeletal defects, in vivo and in vitro, that resemble those of cdc42-1 mutants. Moreover, STE20 overexpression suppresses cdc42-1 growth defects and cytoskeletal defects in vivo, and Ste20 kinase corrects the actin-assembly defects of permeabilized cdc42-1 cells in vitro. Thus, PAKs are effectors of Cdc42 in pathways that regulate the organization of the cortical actin cytoskeleton. PMID- 9742400 TI - Generation of purified neural precursors from embryonic stem cells by lineage selection. AB - Mouse embryonic stem (ES) cells are non-transformed cell lines derived directly from the pluripotent founder tissue in the mouse embryo, the epiblast [1-3]. Aggregation of ES cells triggers the generation of a diverse array of cell types, including neuronal cells [4-7]. This capacity for multilineage differentiation is retained during genetic manipulation and clonal expansion [8]. In principle, therefore, ES cells provide an attractive system for the molecular and genetic dissection of developmental pathways in vitro. They are also a potential source of cells for transplantation studies. These prospects have been frustrated, however, by the disorganised and heterogeneous nature of development in culture. We have therefore developed a strategy for genetic selection of lineage restricted precursors from differentiating populations. Here, we report that application of such lineage selection enables efficient purification of neuroepithelial progenitor cells that subsequently differentiate efficiently into neuronal networks in the absence of other cell types. PMID- 9742401 TI - Src-like adaptor protein (Slap) is a negative regulator of mitogenesis. AB - The Src-like adaptor protein (Slap) is a recently identified adaptor protein containing Src homology 3 (SH3) and SH2 domains. Slap is found in a wide range of cell types and was shown to interact with the Eck receptor tyrosine kinase in a yeast two-hybrid interaction screen [1]. Here, we found that Slap is expressed in NIH3T3 cells and could associate with the activated platelet-derived growth factor (PDGF) receptor. Using mutated versions of the PDGF receptor and phosphopeptide competition experiments, we determined that Slap has the highest affinity for the Src-binding site of the PDGF receptor. Our inability to produce cell lines that stably expressed Slap suggested that Slap inhibited cell growth. We further investigated this issue by transiently expressing Slap by microinjection. Overexpression of Slap by this method inhibited DNA synthesis induced by PDGF and serum, whereas overexpression of the adaptor proteins Grb2 and Shc did not. Finally, microinjection of a Slap antibody into NIH3T3 cells that had been stimulated with suboptimal doses of growth factors potentiated the effects of the growth factors. These data suggest that, unlike other adaptor proteins, Slap is a negative regulator of signalling initiated by growth factors. PMID- 9742402 TI - The lunatic fringe gene is a target of the molecular clock linked to somite segmentation in avian embryos. AB - The most obvious segments of the vertebrate embryo are the trunk mesodermal somites which give rise to the segmented vertebral column and the skeletal muscles of the body. Mechanistic insights into vertebrate somitogenesis have recently been gained from observations of rhythmic expression of the avian hairy related gene (c-hairy1) in chick presomitic mesoderm (PSM), suggesting the existence of a molecular clock linked to somite segmentation ([1]; reviewed in [2]). Here, we show that lunatic Fringe (IFng), a vertebrate homolog of the Drosophila Fringe gene, is also expressed rhythmically in PSM. The PSM expression of IFng was observed as coordinated pulses of mRNA resembling the expression of c hairy1. We show that c-hairy1 and IFng expression in the PSM are coincident, indicating that both genes are responding to the same segmentation clock. The genes were found to differ in their regulation, however; in contrast to c-hairy1, IFng mRNA oscillations required continued protein synthesis, suggesting that IFng could be acting downstream of c-hairy1 in the clock mechanism. In Drosophila, Fringe has been shown to play a role in modulating Notch-Delta signalling [3,4], a pathway which in vertebrates has been implicated in defining somite boundaries [5-9]. These observations place the segmentation clock upstream of the Notch Delta pathway during vertebrate somitogenesis. PMID- 9742403 TI - Essential role of alpha 6 integrins in cortical and retinal lamination. AB - Extracellular matrix (ECM) is believed to play important roles in many aspects of nervous system development [1]. The laminins are ECM glycoproteins expressed in neural tissues and are potent stimulators of neurite outgrowth in vitro [1-3]. Genetic approaches using Drosophila and Caenorhabditis elegans have demonstrated a role for laminin and a laminin receptor in vivo in axon pathfinding and fasciculation, respectively [4,5]. In higher organisms, however, the role of laminins in the development of the nervous system is poorly understood. Integrins alpha 6 beta 1 and alpha 6 beta 4 are major laminin receptors. A role for the alpha 6 integrin in neurulation has been reported in amphibians [6]. We previously described mice lacking integrin alpha 6; these mice died at birth with severe skin blistering [7]. Detailed analyses of integrin alpha 6-/- mice reported here revealed abnormalities in the laminar organization of the developing cerebral cortex and retina. Ectopic neuroblastic outgrowths were found on the brain surface and in the vitreous body in the eye. Alterations of laminin deposition were found in mutant brains. Thus, this study provides evidence for an essential role of integrin-laminin interactions in the proper development of the nervous system. These observations are particularly significant given the recent report that human patients suffering from epidermolysis bullosa can carry mutations in ITGA6, the gene encoding the alpha 6 integrin chain [8,9]. PMID- 9742404 TI - Telencephalic progenitors maintain anteroposterior identities cell autonomously. AB - Grafting experiments have demonstrated that determination of anteroposterior (AP) identity is an early step in neural patterning that precedes dorsoventral (DV) specification [1,2]. These studies used pieces of tissue, however, rather than individual cells to address this question. It thus remains unclear whether the maintenance of AP identity is a cell-autonomous property or a result of signaling between cells within the grafted tissue. Previously, we and others [3-5] have used transplants of dissociated brain cells to show that individual telencephalic precursor cells can adopt host-specific DV identities when they integrate within novel regions of the telencephalon. We have now undertaken a set of transplantations during the same mid-neurogenic period used in the previous studies to assess the ability of telencephalic progenitors to integrate and differentiate into more posterior regions of the neuraxis. We observed that telencephalic progenitors were capable of integrating and migrating within different AP levels of the central nervous system (CNS). Despite this, we found that telencephalic progenitors that integrated within the diencephalon and the mesencephalon continued to express a telencephalic marker until adulthood. We speculate that during neurogenesis individual progenitors are determined in terms of their AP but not their DV identity. Hence, AP identity is maintained cell autonomously within individual progenitors. PMID- 9742405 TI - APA Distinguished Early Career Award to Mark Blumberg. PMID- 9742406 TI - Thermoregulatory competence and behavioral expression in the young of altricial species--revisited. AB - The behavioral and physiological thermoregulatory capabilities of newborn and infant mammals have been studied for over half a century. Psychobiologists have noted that the infants of altricial species (e.g., rats) have physical and physiological limitations such that heat loss overwhelms heat production, thus forcing a reliance on behavioral thermoregulation for the maintenance of body temperature. Recent evidence, however, suggests that a modification of this view is justified. Specifically, throughout a range of moderately cold air temperatures, nonshivering thermogenesis by brown adipose tissue contributes significantly to the infant rat's behavioral and physiological adaptations to cold challenge. Given the prominent use of altricial species for the study of infant behavior, increased understanding of the infant's physiological responses to cold and the effect of thermal factors on behavior is warranted. PMID- 9742408 TI - Bottle-fed neonates prefer an odor experienced in utero to an odor experienced postnatally in the feeding context. AB - The head-orientation response of 2- and 4-day-old bottle-feeding neonates was studied in paired-choice odor tests. Three tests were conducted at Days 2 and 4 after birth to assess the development of the relative response between two salient odors from the prenatal and postnatal environments: (a) amniotic fluid (AF) versus formula milk (FM), (b) FM versus control stimulus (distilled water), and (c) AF versus control stimulus. At both ages, AF and FM elicited positive orientation when presented simultaneously with the control stimulus, indicating that both odors were detectable and attractive to the infants. However, when AF and FM were presented concurrently, the infants expressed significantly longer orientation response toward AF odor than toward FM odor at the age of 2 and 4 days. Within the first 4 days of life, bottle-feeders thus display olfactory preference for a prenatal substrate over a postnatal substrate to which they were recurrently exposed in the feeding situation. PMID- 9742407 TI - Variation in motor activity on different time scales and responsiveness to oral stimulation in the rat fetus. AB - The near-term rat fetus exhibits brief oral grasp responses to discrete presentations of an artificial nipple. In the present experiment, an artificial nipple was presented to individual fetal subjects 10 times. Five of the presentations were timed to occur when spontaneous fetal motor activity was low and five while activity was high, as determined by the baseline activity for the individual fetus. The likelihood of responding to the artificial nipple was increased when the fetus was relatively inactive at the moment of stimulus presentation. Furthermore, stimulus presentations that resulted in oral grasping of the artificial nipple were associated with greater point-to-point variability (2-s intervals) in motor activity during the 30-s period preceding the presentation of the artificial nipple. This pattern of results indicates that the recent history of general motor activity as well as the level of activity at the instant of stimulus presentation may contribute to variation in responding to biologically relevant stimuli early in development. PMID- 9742409 TI - Mechanisms underlying the absence of the pubertal shift in the playful defense of female rats. AB - Due to the action of testicular hormones in the perinatal period, juvenile male rats engage in more play fighting than juvenile females. Also, following puberty, males, but not females, switch to using adultlike defensive tactics more frequently during play. This change in play is also due to the action of testicular hormones perinatally. In this study, two experiments were conducted to determine if the pubertal transition in defense could be induced in females. For Experiment 1, male and female cagemates were tested before and after puberty with familiar and unfamiliar partners. Even when playfully interacting with subadult males, females did not increase the use of the adultlike defensive tactics. For Experiment 2, neonatal females were either injected with testosterone propionate (TP) or ovariectomized (OVX), and again tested before and after puberty. While the TP-treated females had higher frequencies of play fighting, they did not change their pattern of defense following puberty. The OVX females exhibited the lower frequency of play fighting typical of females, but changed their pattern of defense with increased age. Thus, it appears that the pattern of pubertal change in playful defense typical of males is inhibited by ovarian hormones. The mechanisms by which ovarian hormones could exert this effect on developing females are discussed. PMID- 9742410 TI - Odor preferences in neonatal and weanling rats. AB - In order to establish odors which can be used in appetitive and aversive conditioning paradigms, naive rat pups, postnatal day 7 (i.e., PND 7) and weanlings (PND 25), were placed in a rectangular open field with an odorant at one or both ends. Time spent over each odor was measured for 3 min. At both ages subjects avoided peppermint, orange, and lemon odors in favor of fresh home-cage bedding. Comparing any of these three odorants with each other resulted in no significant differences in preferences. In experiments using banana odorant, equal time was spent between banana and no odorant. However, in a two-odorant choice between banana and peppermint, weanlings preferred banana whereas pups showed no preference. The results of this study indicate that in an appetitive learning paradigm, peppermint, orange, or lemon odors may be used, while in aversive learning paradigms banana odor may be more appropriate for weanlings. PMID- 9742411 TI - Negative emotionality and cortisol during adolescent pregnancy and its effects on infant health and autonomic nervous system reactivity. AB - This research examined the relations among maternal emotionality, biology, and infant outcome and autonomic nervous system reactivity (cardiac vagal tone). The sample consisted of 27 pregnant adolescents and their 3-week-old infants. Measures of anxiety, depression, anger, and saliva cortisol were obtained from the adolescents both pre- and postnatally. Infant outcome measures consisted of gestational age at delivery, birth weight, number of risk factors at birth and at 24 hr, Apgar score at 1 and 5 min, abnormalities on newborn physical exam, number of resuscitation measures used on the infant, and cardiac vagal tone. Significant relations were found among the adolescent's emotionality, infant physical outcomes, and cardiac vagal tone. Higher concentrations of adolescent cortisol were associated with lower infant Apgar scores and an increased need for resuscitation measures performed on the infant. The positive association between negative emotions and better infant outcomes also was found and may reflect the sensitivity of the adolescents to their feelings and needs during pregnancy. Social support during pregnancy mediated the effects of maternal negative emotionality and infant cardiac vagal tone. PMID- 9742413 TI - Definitions and terminology of compartment syndrome and Volkmann's ischemic contracture of the upper extremity. AB - Increased tissue pressure within the confines of a nondistensible anatomic compartment increases venous pressure, causes vascular compression, decreases the arteriovenous gradient, and results in a compartment syndrome. The decreased blood flow and hypoxia result in cellular damage of muscles, nerves, and vascular endothelium. Richard von Volkmann's legacy has taught us that a patient's overall status must be monitored to follow the systemic determinants of peripheral blood flow and oxygen transport. Limbs must also be monitored with vigilance. A high index of suspicion must always be present, and compartment pressures can be measured directly to aid in clinical decision making regarding the status of a compartment. The remaining articles in this issue describe and explore von Volkmann's syndrome using the terms and concepts introduced here. PMID- 9742412 TI - Endogenous opioids and the first suckling episode in the rat. AB - Endogenous activity at opioid receptors affects the appetitive behavior of Caesarean-delivered rat pups during presentation of a surrogate nipple that provides milk. Blockade of opioid receptors by peripheral injection of naloxone has no effect on responses evoked by the surrogate nipple. Similarly, blockade of caudal brain opioid receptors by injection of naloxone into the cisterna magna has no effect on the pup's behavior in response to the surrogate nipple. However, blockade of rostral opioid receptors by injection of naloxone into the cerebral ventricles increases the latency to the first oral grasp response, decreases total time on the nipple, and virtually eliminates ingestion of milk from the surrogate nipple (Experiment 1). Blockade of endogenous opioid activity does not affect responses to a nipple that provides distilled water (Experiment 2) or to an empty surrogate nipple (Experiment 3). These data indicate that during the initial suckling episode endogenous opioids in rostral brain regions affect the pup's behavioral responses to the nipple. The results are consistent with the hypothesis that milk engages opioid systems during the first suckling and that endogenous opioids play a role in early suckling. PMID- 9742415 TI - Anatomy of the upper extremity muscle compartments. AB - Multiple and anatomically distinct compartments are present in the upper extremity. A compartment is defined as an enclosed space formed by fascia or by a combination of fascia and bone that contains one or more muscles. Knowledge of these anatomic compartments and their contents will facilitate early diagnosis and treatment. PMID- 9742414 TI - A historical review of compartment syndrome and Volkmann's ischemic contracture. AB - Compartment syndrome has been a recognized disease entity since the mid nineteenth century. Extensive research and clinical observation have allowed for a better understanding of this uniquely complex disease process. Outstanding contributions by clinicians over the past century have provided the basis for our current perspective on its pathogenesis, diagnosis, and treatment. Although Volkmann's early reference to increased "pressure" as the cause of this syndrome was made over 100 years ago, today's modern concept of treatment to prevent contracture based on increased intracompartment pressure has evolved only through the combined efforts of various committed clinicians during the past century. PMID- 9742416 TI - Causes of upper extremity compartment syndrome. AB - Multiple causes, including various types of trauma, prolonged compression, muscle avulsions, burn, snake bites, high-pressure injection injuries, exercise, infection, bleeding, and intravenous drug infiltration, have been reported to lead to the development of a compartment syndrome in the upper extremity. Awareness of the different causes and risks of compartment syndrome should facilitate early diagnosis and prompt treatment to help avoid the development of serious sequelae such as Volkmann's ischemic contracture. A review of the various reports of upper extremity compartment syndromes and classification systems is presented to assist in the understanding of this condition's development. PMID- 9742417 TI - Current concepts in the pathophysiology, evaluation, and diagnosis of compartment syndrome. AB - This article reviews present knowledge of the pathophysiology and diagnosis of acute compartment syndromes. Recent results using compression of legs in normal volunteers provide objective data concerning local pressure thresholds for neuromuscular dysfunction in the anterior compartment. Results with this model indicate that a progression of neuromuscular deficits occurs when IMP increases to within 35 to 40 mm Hg of diastolic blood pressure. These findings provide useful information on the diagnosis and compression thresholds for acute compartment syndromes. Time factors are also important, however, and usually are incompletely known in most cases of acute compartment syndrome. Although the slit catheter is a very good technique for monitoring IMP during rest, these catheters and their associated extracorporeal transducer systems are not ideal. Recently developed miniature transducer-tipped catheters and, perhaps, future development of noninvasive techniques may provide accurate recordings of IMP in patients with acute compartment syndromes. PMID- 9742418 TI - Acute compartment syndrome of the arm. AB - Compartment syndrome of the arm, although uncommon, may result in significant disability if the diagnosis is missed. A high index of suspicion is needed, particularly in those patients at high risk, especially intoxicated or comatose patients. This article discusses the relevant anatomy, pathogenesis, diagnosis, prognosis, and complications of this syndrome. PMID- 9742419 TI - Acute compartment syndrome of the forearm. AB - The forearm is the most common site for compartment syndrome in the upper extremity. The three compartments of the forearm include the volar (anterior or flexor), the dorsal (posterior or extensor), and the mobile wad. Both-bone forearm fractures and distal radius fractures are common initial injuries in adults that lead to acute forearm compartment syndrome. Supracondylar fractures, especially those with associated vascular injuries, are frequent causes of compartment syndrome in children. The flexor digitorum profundus and flexor pollicis longus are among the most severely affected muscles because of their deep location, adjacent to bone. Initial treatment consists of removal of occlusive dressings or splitting or removal of casts. If symptoms do not resolve rapidly, fasciotomy is indicated. Decompression fasciotomy of the forearm is performed through volar or dorsal approaches. The medial nerve is decompressed throughout its course, including high-risk areas deep to the lacertus fibrosus; between the humeral and ulnar heads of the pronator teres, the proximal arch, and deep fascial surface of the flexor digitorum superficialis; and the carpal tunnel. PMID- 9742420 TI - Compartment syndrome of the hand and wrist. AB - High clinical suspicion is of paramount importance in evaluating the hand or wrist for an evolving compartment syndrome. A detailed history coupled with a thorough physical examination form the basis for the diagnosis. The use of techniques to measure compartment pressures forms the objective foundation to assist in formulating the correct treatment plan. The particular technique used to measure the compartments is not critical as long as the information is evaluated in the context of the history and physical examination. No absolute threshold pressure exists over which a fasciotomy is indicated. The need for immediate fasciotomy once the diagnosis is made is clear, however. No one can be faulted for proceeding with a fasciotomy on clinical grounds alone, even when the appearance of findings typically associated with compartment syndrome at surgery (herniating muscle bellies, edema, etc.) are less than convincing. At their worst, the wounds from a fasciotomy present a cosmetic challenge. Great fault can be assigned, however, to the clinician who chooses to ignore an evolving compartment syndrome that unnecessarily places the patient at risk of permanent disability. Here, the cosmetic benefit of avoiding the fasciotomy is overwhelmed by the often-devastating dysfunction created by ischemic damage to the contents of the affected compartments. Once the damage is done, it is permanent. A thorough understanding of the pertinent anatomy is critical to safe, efficacious treatment. Handled promptly and judiciously, compartment syndrome of the hand and wrist can be managed effectively, decreasing the morbidity associated with this potentially devastating and debilitating process. PMID- 9742421 TI - Acute carpal tunnel syndrome. AB - Although the carpal tunnel is open at both ends, it has the physiologic properties of a closed compartment bounded by synovium proximally and distally. When the intracarpal canal interstitial pressure rises above a critical threshold pressure, capillary blood flow is reduced below the level required for median nerve viability. Acute carpal tunnel syndrome is recognized frequently as occurring secondary to wrist trauma and infrequently due to a variety of infectious, rheumatologic, and hematologic disorders. This condition warrants prompt recognition and the treatment is early carpal tunnel release. PMID- 9742422 TI - Compartment syndromes in obtunded patients. AB - A high index of suspicion for a compartment syndrome in the upper extremity should be maintained in all obtunded patients who are at risk for the condition. Obtunded patients are those with a dulled or altered physical or mental status secondary to injury, illness, or anesthesia; those with diminished or absent sensation in the upper extremity because of nerve injury or anesthesia; and those whose ability to communicate is impeded, such as infants and young children and the mentally ill or disabled. These patients represent a vulnerable group whose inability to demonstrate the hallmark symptoms and signs of the syndrome puts them in jeopardy of a late diagnosis of a compartment syndrome and its potentially devastating sequelae. The most likely causes of a compartment syndrome in this population are skeletal or soft-tissue trauma, prolonged limb compression, thrombolytic therapy after myocardial infarction, arterial or intravenous fluid administration, and upper extremity Surgery. Whenever a compartment syndrome of the hand, forearm, or upper arm is suspected, the obtunded patient should be examined closely and frequently, and any changes over time should be documented carefully. Intracompartmental pressure measurement provides a useful adjunct to the physical examination and history in these patients and may be diagnostic if other symptoms and signs are obscured. Once the compartment syndrome is diagnosed, emergent fasciotomy is indicated. To avoid a loss of function in the obtunded patient, special care must be taken postoperatively to assure that early motion exercises are carried out. PMID- 9742423 TI - Crush syndrome of the upper extremity. AB - Crush syndrome is the severe systemic manifestation of prolonged muscle compression and compartment syndrome. Careful patient assessment, early diagnosis, and aggressive treatment are vital to prevent multiorgan failure and death. Medical management of systemic complications, along with operative procedures of fasciotomy and debridement, are indicated with accompanying compartment syndrome. Debridement of necrotic and nonviable tissue is necessary; significant risks of infection and hemorrhage remain until the wounds can be subsequently closed or covered with skin graft. Crush syndrome and muscle necrosis in a closed injury without compartment syndrome may be followed clinically until healing or demarcation of a gangrenous part occurs, providing the patient's general medical condition, including renal function, can be maintained. Fasciotomy and hyperbaric oxygen will not reverse necrosis of muscle in the absence of compartment syndrome and therefore do not affect outcome of the extremity. Overall, prognosis is improved by early diagnosis and treatment, but outcome of the crushed extremity is poor and Volkmann's contracture often results. PMID- 9742425 TI - Upper extremity pediatric compartment syndromes. AB - Fractures are associated with the majority of compartment syndromes in children. Respect for associated soft-tissue injuries and recognition of specific fractures that can put a limb at risk for compartment syndrome are essential for prevention or successful treatment with early decompressive fasciotomies. In those limbs at risk for compartment syndrome, percutaneous pinning or intramedullary fixation provides fracture stabilization and prevents problems noted with standard cast treatment. Any condition that causes increased tissue pressure within a limited space can lead to compartment syndrome, however. It therefore is important to identify the injured, ill, or hospitalized child with unexplained changes in pain status or soft tissues. In particular, the agitated child with increasing analgesia requirements requires a thorough evaluation. The child's behavior should not be attributed to young age, fear, or fracture pain. This is a trap that must be avoided to prevent the disastrous outcomes of a missed compartment syndrome. PMID- 9742426 TI - Exertional compartment syndrome of the upper extremity. AB - Exertional compartment syndrome is characterized by intracompartmental pressures that rise transiently following repetitive motion or exercise, thereby producing temporary, reversible ischemia, pain, weakness, and, occasionally, neurologic deficits. The exact cause or pathogenesis remains unclear; a disturbance of microvascular flow caused by elevated intramuscular pressure leads to tissue ischemia, depletion of high-energy phosphate stores, and cellular acidosis. Anatomic contributing factors may include a limited compartment size, increased intracompartmental volume, constricted fascia, loss of compartment elasticity, poor venous return, or increased muscle bulk. The diagnosis is suspected based on history and confirmed with physical examination and intramuscular pressure evaluation before and after exercise (stress test). Differential diagnosis includes claudication or other vascular abnormalities, myositis, tendinitis, periostitis, chronic strains or sprains, stress fracture, other compression or systemic neuropathies, and cardiac abnormalities with angina or referred extremity pain. Initial treatment includes activity modification; refractory symptoms can be managed with elective fasciotomy. PMID- 9742424 TI - The role of prophylactic fasciotomy and medical treatment in limb ischemia and revascularization. AB - Multiple studies have demonstrated that muscle poorly tolerates ischemia. When the ischemic state is unduly prolonged, the successfully replanted or revascularized limb undergoes deleterious biochemical reactions that cascade to vessel intimal damage, increased vessel permeability, and lowering of pH. The resultant tissue edema leads to increasing compartment pressures, which not only impede the recovery of function, but also can lead to irreversible muscle necrosis, increased risk of infection, and sepsis if not reversed in a timely fashion. The development of compartment syndrome jeopardizes not only the injured limb, but life itself secondary to the biochemical toxins produced by the ischemic extremity. A thorough understanding of the biochemistry of ischemia and reperfusion provides insight into the role of fasciotomy in the replanted or revascularized extremity. The scientific basis for fasciotomy in the revascularized or replanted limb is discussed as well as the potential "protective" role of pharmacologic agents in ischemic and reperfusion injury. PMID- 9742427 TI - Volkmann's ischemic contracture of the upper extremity. AB - Upper extremity deformity of ischemic contracture usually includes elbow flexion, forearm pronation, wrist flexion, thumb flexion and adduction, digital metacarpophalangeal joint extension, and interphalangeal joint flexion. Treatment of mild contractures consists of either nonoperative management with a comprehensive rehabilitation program (to increase range of motion and strenght) or operative management consisting of infarct excision or tendon lengthening. Treatment of moderate-to-severe contractures consists of release of secondary nerve compression, treatment of contractures (with tendon lengthening or recession), tendon or free-tissue transfers to restore lost function, and/or salvage procedures for the severely contracted or neglected extremity. PMID- 9742428 TI - Reconstruction of intrinsic hand deformities. AB - Intrinsic muscle contractures are a frequently overlooked cause of hand dysfunction. Tightness of these muscles may occur despite appropriate management. This article addresses the evaluation and treatment of these contractures. PMID- 9742429 TI - Overview of alternatives for clinical integration. PMID- 9742430 TI - Physician practice management companies: selling your practice. PMID- 9742431 TI - Alternatives for clinical integration: antitrust law considerations. PMID- 9742432 TI - Physician practice integration and consolidation: an antitrust enforcement perspective. PMID- 9742433 TI - Overview of quality measurement and improvement. PMID- 9742434 TI - Cataract outcomes assessment in the general ophthalmology practice. PMID- 9742435 TI - Clinical practice guidelines. PMID- 9742436 TI - Understanding the American Academy of Ophthalmology's preferred practice pattern for cataract. PMID- 9742437 TI - Understanding the new primary open-angle glaucoma preferred practice pattern. PMID- 9742438 TI - Introduction to capitation. PMID- 9742439 TI - Developing capitation rates. PMID- 9742440 TI - Negotiating strategies for capitation. AB - The appropriate strategy to employ in contract negotiations will vary, depending on a number of important factors. These include the relative size and power of the network, the conditions of the local market, the strength of the managed-care payer in the market, and a host of other issues. The negotiating strategy ultimately adopted will be in accordance with the organizations overall preference, style, and needs. In approaching any contract negotiation, two key points should be kept in mind. First, networks must recognize they can become more prepared and empowered for contract negotiations through the acquisition of additional information. Second, form contracts can be changed; nothing is set in stone. Despite the frequent statements of payer organizations that the form cannot be modified to meet the provider's unique desires and needs, in most cases a contract can be modified in one way or another to meet the parties' mutual needs and desires. Everything is negotiable: Any party involved in such negotiations should be positive, avoiding conflict; use a problem-solving approach; agree whenever possible--strive to become "we"; acknowledge the payer's interest and position, expressing a desire to compromise; and acknowledge the authority and ability of the assigned negotiator. Once the managed-care contract is consummated, the network should attempt to ensure that the parties get maximum value from the agreement. One step to achieving such a desired end is to identify a leader who will be responsible for overseeing contract implementation and performance and responsible for knowing the requirements of the agreement inside out and for focusing on all other requirements of the managed-care program. PMID- 9742441 TI - Non-capitation contract evaluation: a primer. PMID- 9742442 TI - Fighting fire with fire: using expert warriors on the managed-care battlefield. PMID- 9742443 TI - Professional liability in the managed-care setting. PMID- 9742445 TI - Accessibility of T-tubule vacuoles to extracellular dextran and DNA: mechanism and potential application of vacuolation. AB - In addition to its function in excitation-contraction coupling, the ability of the T-system of skeletal muscle fibres to undergo reversible vacuolation indicates that it may play a role in volume regulation. The mechanism of reversible vacuolation has been investigated by confocal microscopy using fluorescein dextran to probe the accessibility of T-tubules to the extracellular environment. Vacuolation was induced by loading the fibres with 60-100 nM glycerol for 30 minutes and then returning them to glycerol-free medium. Devacuolation was subsequently induced by the reentry of glycerol. During their formation from T-tubules, the vacuoles filled with fluorescent dextran from the extracellular medium. The inaccessibility of the vacuoles to extracellular ferritin observed in a previous study raised the possibility that the vacuoles may be detached from the surface membrane after their formation. However, it is apparent from the present work that, although the tubules of the treated fibres are constricted, the vacuoles maintain a open connection with the external medium for smaller macromolecules. In the light of these experiments, it is proposed that vacuolation is caused by water moving into T-tubules from the cytoplasm faster than it can exit to the extracellular space during a decrease in fibre volume. Since T-tubules have been implicated in the transfection of skeletal muscle by direct injection, the accessibility of plasmid DNA to T-tubules has also been investigated. DNA penetrated into the vacuoles from the extracellular medium less well than dextran, but many vacuoles containing fluorescent DNA were observed in the superficial layers of vacuolated fibres, and it is suggested that T-tubule vacuolation might be used to improve the efficiency of the transfection of skeletal muscle by direct injection. PMID- 9742446 TI - The tubular vacuolation process in amphibian skeletal muscle. AB - The exposure of amphibian muscle to osmotic shock through the introduction and subsequent withdrawal of extracellular glycerol causes 'vacuolation' in the transverse tubules. Such manoeuvres can also electrically isolate the transverse tubules from the surface ('detubulation'), particular if followed by exposures to high extracellular [Ca2+] and/or gradual cooling. This study explored factors influencing vacuolation in Rana temporaria sartorius muscle. Vacuole formation was detected using phase contrast microscopy and through the trapping or otherwise of lissamine rhodamine dye fluorescence within such vacuoles. The preparations were also examined using electron microscopy, for penetration into the transverse tubules and tubular vacuoles of extracellular horseradish peroxidase introduced following the osmotic procedures. These comparisons distinguished for he first time two types of vacuole, 'open' and 'closed', whose lumina were respectively continuous with or detached from the remaining extracellular space. The vacuoles formed closed to and between the Z-lines, but subsequently elongated along the longitudinal axis of the muscle fibres. This suggested an involvement of tubular membrane material; the latter appeared particularly concentrated around such Z-lines in the electron-micrograph stereopairs of thick longitudinal sections. 'Open' vacuoles formed following osmotic shock produced by extracellular glycerol withdrawal from a glycerol loaded fibre at a stage when one would expect a net water entry to the intracellular space. This suggests that vacuole formation requires active fluid transport into the tubular lumina in response to fibre swelling. 'Closed' vacuoles only formed when the muscle was subsequently exposed to high extracellular [Ca/+] and/or gradual cooling following the initial osmotic shock. Their densities were similar to those shown by 'open' vacuoles in preparations not so treated, suggesting that both vacuole types resulted from a single process initiated by glycerol withdrawal. However, vacuole 'closure' took place well after formation of 'open' vacuoles, over 25 min after glycerol withdrawal. Its time course closely paralleled the development of detubulation reported recently. It was irreversible, in contrast to the reversibility of 'open' vacuole formation. These findings identify electrophysiological 'detubulation' of striated muscle with 'closure' of initially 'open' vacuoles. The reversible formation of open vacuoles is compatible with some normal membrane responses to some physiological stresses such as fatigue, whereas irreversible formation of closed vacuoles might only be expected in pathological situations as in dystrophic muscle. PMID- 9742444 TI - Troponin T: genetics, properties and function. AB - Troponin T (TnT) is present in striated muscle of vertebrates and invertebrates as a group of homologous proteins with molecular weights usually in the 31-36 kDa range. It occupies a unique role in the regulatory protein system in that it interacts with TnC and TnI of the troponin complex and the proteins of the myofibrillar thin filament, tropomyosin and actin. In the myofibril the molecule is about 18 nm long and for much its length interacts with tropomyosin. The ability of TnT to form a complex with tropomyosin is responsible for locating the troponin complex with a periodicity of 38.5 nm along the thin filament of the myofibril. In addition to it structural role, TnT has the important function of transforming the TnI-TnC complex into a system, the inhibitory activity of which, on the tropomyosin-actomyosin MgATPase of the myofibril, becomes sensitive to calcium ions. Different genes control the expression of TnT in fast skeletal, slow skeletal and cardiac muscles. In all muscles, and particularly in fast skeletal, alternative splicing of mRNA produces a series of isoforms in a developmentally regulated manner. In consequence TnT exists in many more isoforms than any of the other thin filament proteins, the TnT superfamily. Despite the general homology of TnT isoforms, this alternative splicing leads to variable regions close to the N- and C-termini. As the isoforms have slightly different effects on the calcium sensitivity of the actomyosin MgATPase, modulation of the contractile response to calcium can occur during development and in different muscle types. TnT has recently aroused clinical interest in its potential for detecting myocardial damage and the association of mutations in the cardiac isoform with hypertrophic cardiomyopathy. PMID- 9742447 TI - Microfluorometric analyses of glycogen in freshly dissected, single skeletal muscle fibres of the cane toad using a mechanically skinned fibre preparation. AB - The main objective of this study was to analyse glycogen in single muscle fibres, using a recently developed microfluorometric method which detects subpicomol amounts of NADPH, glucose and glycogen (as glucosyl units) (detection limit 0.16 0.17 pmol in a 25 nl sample) without fluorochrome amplification. The fibres were freshly dissected from the twitch region of the iliofibularis muscle of the cane toad (Bufo marinus), and were mechanically skinned under paraffin oil to gain access to the intracellular compartments. The results show that (1) glycogen concentrations in toad skeletal muscle fibres range between 25.8 and 369 mmol glucosyl units/litre fibre volume; (2) there is a large variation in glycogen content between individual fibres from the iliofibularis muscle of one animal; (3) there are seasonal differences in the glycogen content of toad single muscle fibres; (4) the total amount of glycogen in single muscle fibres of the toad does not decrease significantly when storing the tissue, under paraffin oil, at 20-25 degree C for up to 6 h or at 4 degree C for up to 24 h; and (5) 15-26% of fibre glycogen can be washed in an aqueous solution at pH 5-7, within 5 min, while 74 85% of fibre glycogen remains associated with the washed skinned fibre, even after 40 min exposure of the skinned fibre preparation to the aqueous environment. The retention of most glycogen in the fibre preparation after mechanical removal of the plasma membrane and extensive washing indicates that in toad skeletal muscle fibres the largest proportion of glycogen is tightly bound to intracellular structures. The results also show that the skinned muscle fibre preparation is well suited for microfluorometric glycogen determination, since low molecular weight non-glycogen contributors to the fluorescence signal can be removed from the myoplasmic space prior to the glycogen hydrolysis step. PMID- 9742448 TI - Generation of functional beta-actinin (CapZ) in an E. coli expression system. AB - beta-actinin (CapZ) is a heterodimeric actin-binding protein which caps the barbed end of action filaments and nucleates actin-polymerization in a Ca2+ independent manner. In myofibrils it is localized in the Z-lines. As judged by these properties of b-actinin, it is conceivable that beta-actinin is involved in the regulation of actin assembly, especially in the formation of I-Z-I complex during myofribrillogenesis. In this study, we devised a system to produce functional beta-actinin in E. Coli. The cDNAs of beta I' and beta II subunits of beta-actinin were obtained by RT-PCR methods using the published sequence as references, and subcloned in a pET vector. When the proteins were produced with the cDNA of either beta I' and beta II in E. coli, the proteins were insoluble and non-functional. However, when the cDNAs encoding the two subunits were cloned into a single vector and both proteins were expressed simultaneously, the proteins became soluble and purified as a functional heterodimer The activity of the purified proteins was not distinguishable from that of beta-actinin purified from skeletal muscle. PMID- 9742449 TI - A model for the function of the bisphosphorylated heart-specific troponin-I N terminus. AB - Bisphosphorylation of two adjacently located serine residues in the heart specific N-terminus of the cTnl subunit reduces calcium affinity of the cTnC subunit. An interaction of the phosphorylation region of cTnI with acidic residues of another cTn subunit has been proposed formerly based on 31P nuclear magnetic resonance (NMR) data. A possible candidate is cTnC. Thus, an interaction model of cTnC with the bisphosphorylated cTnI N-terminus has been built using a homology model of hcTnC based on the crystal structure of tusTnC and the structure of the phosphorylation region of cTnI determined by 2D NMR. By computational search, five cluster of acidic residues of cTnC might interact with the cTnI phosphorylation region. Three sites could be excluded by 31P-NMR experiments. The two remaining sites are located in the N-terminal helix of cTnC and between calcium binding sites III and IV. Reorientation of the arginine and phosphoserine sidechains within the phosphorylation region as proposed by refined docking could explain the formerly measured changes in pKaapp values. Thus, local pKa changes might lead to the reduction of calcium affinity observed upon cTnI bisphosphorylation. PMID- 9742450 TI - Comparison of sarcoplasmic reticulum capabilities in toadfish (Opsanus tau) sonic muscle and rat fast twitch muscle. AB - The sonic muscle of the oyster toadfish, Opsanus tau, can produce unfused contractions at 300 Hz. Electron microscopy shows a great abundance of the Sarcoplasmic reticulum (SR) in this muscle, but no functional characterization of the capabilities of the SR has been reported. We measured the oxalate-supported Ca2+ uptake rate and capacities of homogenates of toadfish sonic muscle and rat extensor digitorum longus (EDL) muscle, and estimated the number of pump units by titration with thapsigargin, a high-affinity, specific inhibitor of the SR Ca ATPase. The Ca2+ uptake rate averaged 70.9 +/- 9.5 mumol min -1 per g tissue for the toad fish sonic muscle, and 73.5 +/- 3.7 mumol min -1 g-1 for rat EDL. The capacity for Ca2+ -oxalate uptake was 161 +/- 20 mumol g -1 and 33 +/- 2 mumol g 1 for toadfish sonic muscle and rat EDL, respectively. Thus, the rates of Ca2+ uptake were similar in the two muscles, but the toadfish sonic muscle had about five times the capacity of the rat EDL. The number of pumps as estimated by thapsigargin titration was 68 +/- 4 nmol of Ca-ATPase per g tissue in the toadfish, and 42 +/- 5 nmol Ca-ATPase per g tissue in the rat EDL. The turnover number, defined as the Ca2+ uptake divided by the number of pumps, was 1065 +/- 150 min -1 for toadfish and 1786 +/- 230 min -1 for rat EDL (p < 0.05) at 37 degrees C. The Ca2+ uptake rate of toadfish sonic muscle at 22 degree C, a typical temperature for calling toadfish, averaged 42 +/- 1% of its rate at 37 degree C. At these operating temperatures, the toadfish SR is likely to be slower than the rat fast-twitch SR, yet the toadfish sonic muscle supports more rapid contractions. One explanation for this is that the voluminous SR provides activator Ca2+ for contraction, but the abundant parvalbumin plays a major role in relaxation. PMID- 9742451 TI - Calcium handling by the sarcoplasmic reticulum during oscillatory contractions of skinned skeletal muscle fibres. AB - Isometric ATP consumption and force were investigated in mechanically skinned fibres from iliofibularis muscle of Xenopus laevis. Measurements were performed at different [Ca2+], in the presence and absence of caffeine (5 nM). In weakly Ca2+-buffered solutions without caffeine, spontaneous oscillations in force and ATPase activity occurred. The repetition frequency was [Ca2+]-and temperature dependent. The Ca2+ threshold (+/- SEM) for the oscillations corresponded to a pCa of 6.5 +/- 0.1. The maximum ATP consumption associated with calcium uptake by the sarcoplasmic reticulum (SR) reached during the oscillations was similar to the activity under steady-state conditions at saturating calcium concentrations in the presence of caffeine. Maximum activity was reached when the force relaxation was almost complete. The calculated amount of Ca2+ taken up by the SR during a complete cycle corresponded to 5.4 +/ 0.4 mmol per litre cell volume. In strongly Ca2+-buffered solutions, caffeine enhanced the calcium sensitivity of the contractile apparatus and, at low calcium concentrations, SR Ca uptake. These results suggest that when the SR is heavily loaded by net Ca uptake, there is a massive calcium-induced calcium release. Subsequent net Ca uptake by the SR then gives rise to the periodic nature of the calcium transient. PMID- 9742452 TI - Effects of pre- and perinatal exposure to hypergravity on muscular structure development in rat. AB - This study evaluated the influence of precocious exposure to hypergravity on the expression of myosin heavy chain (MHC) protein isoforms in nape, masticatory and respiratory developmental rat muscles. Pregnant females were maintained at 1.8 g from the 11th day of gestation to the 7th day after birth. The 7-day-old rats were used for muscle sampling. Hypergravity induced a marked decrease in the weight and protein content of all six muscles. Three MHC isoforms were detected in the young rats' muscles: embryonic (E), perinatal (P) and slow type 1 MHC. In centrifuged nape and masticatory muscles, there was a decrease in MHC E and an increase in P without reduction (indeed, even an increase) in MHC 1, whereas in the respiratory muscle MHC E was increased and MHC 1 decreased. These results indicate that hypergravity produces important changes in the contractile properties not only of antigravity muscles but also masticatory and respiratory muscles. MHC P has a higher shortening velocity than MHC E, which has a higher one than MHC 1. The hypergravity-induced transformations of MHC isoforms would thus lead to increased velocity of all muscles studied. In spite of the observation of a hypergravity-induced muscle hypotrophy, the results of this study reflect the adaptational properties of developing muscles to increased gravitational forces. PMID- 9742453 TI - h1- and h2-calponins are not essential for norepinephrine- or sodium fluoride induced contraction of rat aortic smooth muscle. AB - To investigate the controversial issue concerning the role of calponin in smooth muscle contraction, this study examined the relationship between smooth muscle calponin and the contraction of aortic rings from different strains of rats: Sprague-Dawley (SD), Wistar, and Wistar Kyoto (WKY). Western blot analysis demonstrated that h1- and h2-calponins are present in aortic smooth muscle from adult SD rats but not Wistar or WKY rats. Nevertheless, h1-calponin is detectable in stomach from Wistar rats, although at a much lower level compared with that in the SD rat stomach. This suggests that a repressed expression of the gene, instead of a simple null mutation, may have caused its absence from the aortic smooth muscle. Despite the presence or absence of calponin, the aortic smooth muscles from the different strains of rats all develop contractions in response to the physiological agonist norepinephrine (NE) and following activation with the plasma membrane receptor-independent NaF induction. The data indicate that h1 and h2-calponins are not essential for NE- and NaF-induced contractions in aortic smooth muscle. The calponin-positive adult SD rat aorta was found to be more sensitive in contractile response to NE and NaF inductions compared with the calponin-negative rat aortae. This may imply a potential modulator function of calponin in the contraction of smooth muscle, whereas other contractile protein isoform differences between these rat strains may also play a role. PMID- 9742454 TI - Amino-acid sequence of squid myosin heavy chain. AB - This work describes the determination of the cDNA sequence encoding the myosin heavy chain (MHC) of the squid, Loligo pealei. To date, the amino-acid sequence of the MHC of calcium-regulated myosins is known only for two closely related species of scallops. We have determined the sequence of the entire coding region of the muscle MHC of squid, a cephalopod, and compared it with the MHC of scallops, which are pelecypods, and to other regulated and non-regulated myosins. Residues present in the MHC of only regulated myosins have been identified. The 6504 base pair (bp) sequence contains an open reading frame of 5805 nucleotides, which encodes 1935 amino acids. The sequence includes 697 bps of 3' untranslated sequence and 2 bps of 5' untranslated sequence. The deduced amino-acid sequence shows the squid MHC to be 72-73% identical and 86-87% similar to the calcium regulated scallop MHCs cloned previously. In contrast, the squid MHC sequence is only 54-55% identical and 74% similar to skeletal MHCs of non-regulated myosins such as human fast skeletal embryonic and human perinatal skeletal muscle, and 39 40% identical and 60-62% similar to smooth muscle MHC of rabbit uterus muscle and chicken gizzard muscle, respectively. We have also detected two isoforms of the MHC in squid that appear to be spliced variants of a single myosin gene. These isoforms differ in the sequence encoding the surface loop at the nucleotide binding site. Taken together, our data may help to identify more precisely the residues that are crucial in regulated myosins. PMID- 9742455 TI - Comments on the paper by Dr. David Smith entitled "A strain-dependent ratchet model for [phosphate]- and [ATP]-dependent muscle contraction". PMID- 9742456 TI - In vitro and in vivo prolonged biological activities of novel vitamin C derivative, 2-O-alpha-D-glucopyranosyl-L-ascorbic acid (AA-2G), in cosmetic fields. AB - The biological activity of the novel vitamin C derivative, 2-O-alpha-D glucopyranosyl-L-ascorbic acid (AA-2G), was evaluated in vitro and in vivo. The percutaneous absorption of AA-2G was determined in five Japanese males. The excretion of ascorbic acid (AA) in the subjects administered AA-2G was sustained for a longer period than in the subjects administered ascorbic acid 2-phosphate (AA-2P), which is a conventional vitamin C derivative. An analysis of the distribution of AA in the skin showed that small black specks assumed to be AA were observed in the epidermis even 3 d after applying AA-2G. The melanin synthesis in B16 melanoma cells was inhibited more by AA-2G than by AA-2P, and AA 2G also prevented more UV-induced damage of human skin keratinocytes and fibroblasts than AA-2P did. From these in vivo and in vitro results, it is supposed that the conversion of AA-2G to AA is sustained for a long time compared with that of AA-2P, and that AA-2G is an effective and available compound having vitamin C activity in human subjects. PMID- 9742457 TI - Trienzyme treatment for food folate analysis: optimal pH and incubation time for alpha-amylase and protease treatment. AB - Recent reports have indicated that trienzyme treatment before folate determination is essential to obtain the proper folate content in foods. Trienzyme treatment is performed by using alpha-amylase and protease for folate extraction from carbohydrate and protein matrices, and folate conjugase for the hydrolysis of polyglutamyl folates. We evaluated the conditions of pH and incubation time for the treatment with alpha-amylase and protease. Four food items, including fresh beef, white bread, cow's milk, and fresh spinach, were selected for this investigation. We found that optimal pHs for alpha-amylase treatment of beef and cow's milk were 7.0 and 5.0, respectively, whereas those for white bread and spinach were not distinctive at pHs from 2.0 to 7.0. The optimal incubation time for alpha-amylase was 4 h for fresh beef and cow's milk, whereas no distinctive optimal incubation period was found for white bread and fresh spinach. Our data indicate that the conditions for enzyme treatments vary depending on food items. Trienzyme treatment resulted in an increase of more than 50% in the mean folate content over folate conjugase treatment alone. It is necessary to treat food samples with not only traditional folate conjugase, but also with alpha-amylase and protease before folate determination to obtain the actual folate content. PMID- 9742458 TI - Effect of acarbose (alpha-glucosidase inhibitor) on disaccharase activity in small intestine in KK-Ay and ddY mice. AB - The hypoglycemic effect and the alpha-glucosidase activity inhibition of acarbose (AC:alpha-glucosidase inhibitor) were investigated in normal and KK-Ay mice, an animal model of noninsulin-dependent diabetes mellitus (NIDDM). AC improved hyperglycemia after an oral administration of maltose or sucrose, dose dependently in normal mice (1, 10, and 50mg/kg body weight) and in KK-Ay mice (50mg/kg). Furthermore, AC (50mg/kg) significantly inhibited maltase and sucrase activities in the small intestines of normal and KK-Ay mice (inhibitory efficacy: sucrase > maltase). The enzymatic inhibition in KK-Ay mice is stronger than in normal mice. However, AC (50 mg/kg) did not suppress the blood glucose in oral lactose tolerance and did not inhibit the lactase activity in either normal or KK Ay mice. These findings indicate that the AC effect on the inhibition of alpha glucosidase activity is selective for sucrase and maltase in normal and NIDDM mice. PMID- 9742459 TI - Microencapsulated ferrous sulfate to fortify cow milk: absorption and distribution in mice. AB - To determine the absorption and biodistribution of iron from microencapsulated ferrous sulfate (SFE-171), used to fortify dairy products with iron, a comparative study in four groups of 30 mice each was carried out. In two of the groups, the absorption of iron from ferrous ascorbate in water (13.3 +/- 4.3%) and from ferrous sulfate in water (12.7 +/- 3.9%) was determined and taken as reference standards. In the third group the iron absorption from SFE-171 in milk was determined, giving a value of 12.1 +/- 4.2%, which statistically does not differ from the data obtained with either reference standard. In the fourth group, the absorption of iron from ferrous sulfate in milk showed a value of 7.7 +/- 3.4%, which statistically differs with a p < 0.01 from the data corresponding to the other three groups. The biodistribution studies showed that the iron from the SFE-171 follows the same metabolic pathway as the iron from the reference standards thus, giving a higher radioactivity percentage and radioactivity concentration in organs or systems, principally blood, that are closely related to iron metabolism. Our studies allow us to conclude that the iron from SFE-171 in milk follows the same behavior as the nonhemic iron, with a higher absorption than that of ferrous sulfate in milk. PMID- 9742460 TI - Risk factors for the prevalence of malnutrition among urban children in Ghana. AB - A case-control study was completed at the Princess Marie Louise Hospital in Accra, Ghana, to identify risk factors for the prevalence of underweight and severe malnutrition in urban African children. A total of 170 children, aged 8 to 36 mo, with normally nutritional status (> or = 80% W/A of NCHS reference), underweight (moderate malnutrition) (60-80% W/A), or severe malnutrition (< 80% W/A and presence of edema, or < 60% of W/A) were recruited at the clinical ward and at the public health service section of the hospital. Anthropometric measurements and physical examinations were completed, and the guardians were interviewed about their children's health status, birth weight, child care, and household conditions. The severely malnourished children were more likely to have young mothers (p < 0.05) and low weight at birth (p < 0.05). The underweight children were also observed to have low birth weight (p < 0.05). The severely malnourished group showed the tendencies of less feeding frequency (p < 0.01), less access to breast-feeding (p < 0.01), and less support by both parents (p < 0.05). Moreover, the parents of the severely malnourished children had lower educational levels and lower income jobs, compared with those of the normal children (mother's education, p < 0.001; father's education, p < 0.001; mother's occupation, p < 0.05; father's occupation, p < 0.001). No significant differences in most variables existed between the normal and underweight groups. Multivariable analysis resulted in the conclusion that the Z-score of weight-for age, birth weight, and mother's educational level were highly associated with one another. We conclude that low birth weight is one of the important risk factors for the prevalence of underweight and severe malnutrition and that the lack of a mother's education is also a risk factor for the prevalence of severe malnutrition in the urban children in Ghana. PMID- 9742461 TI - Modulation of bone mass and turnover in growing rats by voluntary weight-bearing exercise and glucose supplementation. AB - Female Sprague-Dawley rats, 9 weeks of age, were assigned to four groups: Group 0 (n = 8) was dissected for base-line control, and the other three groups were fed for 3 mo: Group 1 (n = 9), sedentary controls; Group 2 (n = 6), running rats housed in a cage with a treadmill and pair-fed with Group 1; and Group 3 (n = 7), running rats, pair-fed and allowed free access to additional glucose. The distances of voluntary running did not significantly differ between Groups 2 and 3. Menstrual cycles in these rats were apparently maintained as observed from daily running distances. The amount of glucose taken by rats in Group 3 was 3.5 +/- 0.4 (mean and SE) g/d. Body weight (BW) at the end of the experiment for Groups 1, 2, and 3 were 295.0 +/- 7.9, 211.7 +/- 5.4 (p < 0.001 vs. Group 1), and 259.0 +/- 3.5 g (p < 0.01 vs. Group 2), respectively. The parameters of bone mass such as ash weights of the femur and bone mineral content of the lumbar spine and the tibia in Groups 1 and 2 did not differ, but the values were significantly greater in Group 3 than in Group 2. However, these parameter values corrected for BW were significantly greater in Group 2 than in Group 1 and did not significantly differ between Groups 2 and 3. The parameters of bone formation, such as serum bone alkaline phosphatase activity levels and trabecular bone formation rates corrected for BW, were significantly greater in Group 2 than in Group 1 but did not differ between Group 2 and 3. However, the parameters of bone resorption, such as serum tartrate resistant acid-phosphatase levels, were significantly less in Group 3 than in Group 2. These results suggest that voluntary running augments the age-dependent increase in bone mass by modulating the bone turnover when an adequate energy source is supplied under conditions of normal menstruation, and an adequate supply of energy could be necessary to enhance the age-dependent increase in bone mass. PMID- 9742462 TI - Degradation of konjac glucomannan by enzymes in human feces and formation of short-chain fatty acids by intestinal anaerobic bacteria. AB - Konjac (konnyaku) glucomannan was examined for its degradation in human intestines and fermentation products. The konjac glucomannan was degraded almost 100% by soluble enzymes in human feces to give 4-O-beta-D-mannopyranosyl-D mannopyranose (beta-1,4-D-mannobiose), 4-O-beta-D-glucopyranosyl-D-glucopyranose (cellobiose), 4-O-beta-D-glucopyranosyl-D-mannopyranose, and small amounts of glucose and mannose. These three disaccharides were further degraded by a cell associated enzyme(s) to glucose or mannose, or to both. Konjac glucomannan underwent fermentation by intestinal anaerobic bacteria and produced formic acid, acetic acid, propionic acid, and 1-butyric acid. These fatty acids were different in their proportions among test subjects, their total amounts ranging from 17.1% to 48.8% of the initial konjac glucomannan. PMID- 9742463 TI - Bioavailability of magnesium contained in roasted and ground soybean (kinako) as evaluated by serum and bone magnesium contents, kidney calcification, and magnesium absorption. AB - The bioavailability of roasted and ground soybean (kinako) magnesium (Mg) in Fischer 344 strain male rats with respect to serum Mg level, bone Mg contents, kidney calcification, and Mg absorption was evaluated. Four-week-old male rats were divided into four groups of six rats each. The four groups were the control (20SC), Mg-deficient diet (1/3 Mg20SC), 20SCK diet, and 20SCDK diet. The 20SCK and 20SCDK diets were prepared to contain amounts of Mg equal to that in the 20SC diet with kinako or defatted kinako as the Mg source, respectively. After a 4 week experimental period, rats were decapitated and blood serum, right femur, and right kidney were collected, and Mg, calcium (Ca), and phosphorus (P) concentration in those tissues were determined. The Mg balance was also investigated. The serum Mg concentration in the 1/3 Mg20SC group was half the level in the 20SC group, and the serum Ca concentration was higher than in the 20SC group, indicating apparent hypercalcemia. Serum Mg and Ca concentrations in the 20SCK and 20SCDK groups did not significantly differ from those in the 20SC group. Femur Mg concentration in the 1/3 Mg20SC group was lower than in the 20SC group. Femur Mg concentrations in the 20SCK and 20SCDK groups were lower than in the 20SC group, but significantly higher than in the 1/3 Mg20SC group. The kidney Ca concentrations in the 20SCK and 20SCDK groups were significantly higher than those in the 20SC and 1/3 Mg20SC groups, and there was also kidney calcification. These results indicated that kinako and defatted kinako Mg were used effectively as a serum and femur Mg source, but that kinako and defatted kinako carry a risk of kidney calcification when used as the only Mg source. PMID- 9742464 TI - Preparation and characterization of micellar calcium phosphate-casein phosphopeptide complex. AB - Micellar calcium phosphate (MCP) in bovine milk was separated as the complex with casein phosphopeptide (CPP) by the following procedures. Rennet curd obtained from skim milk was suspended in water, the pH was adjusted to 4.6, and the suspension was centrifuged at 1,000 x g. CPP was separated from the precipitated casein by tryptic hydrolysis and ethanol precipitation. The supernatant, which contained calcium and inorganic phosphate liberated from casein micelles by acidification, and CPP were mixed; the pH was adjusted to 6.7; and then the solution was lyophilized. From 1 L of skim milk, 3.16 g of the MCP-CPP complex was obtained. The dried MCP-CPP complex was easily dissolved in water and contained 12.7% calcium, 0.3% magnesium, 3.4% inorganic phosphorous, and 2.2% organic phosphorous. No crystal structure of hydroxyapatite was shown in the MCP CPP complex by the X-ray diffraction analysis, although the pattern of NaCl crystal was observed. The X-ray diffraction pattern of commercial whey mineral, which was prepared by precipitation at alkaline pH from rennet whey, was similar to that of hydroxyapatite. It was confirmed by high-performance gel chromatographic analysis that the form of calcium phosphate in the MCP-CPP complex was similar to that of casein micelles. The MCP-CPP complex was also separated from commercial rennet casein. The method for the separation of MCP-CPP complex described above can be applied to the large-scale preparation. PMID- 9742465 TI - Ability of endogenous folate from soy protein isolate to maintain plasma homocysteine and hepatic DNA methylation during methyl group depletion in rats. AB - We investigated different means of achieving methyl depletion by feeding weanling rats modified AIN diets depleted of folate (FD), folate + choline (FCD), and folate + choline + methionine (FCMD), and examined the consequent effects on folate status, homocysteine levels, and methylation status. Control rats were fed a 12% protein diet consisting of either casein or soy protein isolate (SPI) and containing 2 mg/kg folate, 0.2% choline, and 0.4% methionine. After the rats had been on the diets for 4 and 8 weeks, whole blood folate concentration was measured and found to be significantly depleted in the folate deficient treatments compared with controls at 4 weeks (p < 0.001), this reduction being significantly greater (p < 0.03) in casein-fed rats (60%) than in SPI-fed rats (32%). The omission of choline and methionine from the diet had no further influence on whole blood folate. A significant inverse correlation was observed in the casein-fed rats after 8 weeks between mean plasma homocysteine concentration and decreasing methyl content of the diet (r2 = 0.978, p < 0.002), an effect not seen in the corresponding SPI-fed rats. Hypomethylation of hepatic DNA evidenced by a reduction in 5-methylcytosine content was present in the casein rats fed FCD and FCMD relative to control (p < 0.05). No hepatic DNA methylation changes were observed in the SPI-fed rats. The results obtained in the present work demonstrate that a soy-based diet can compensate against methyl group depletion by maintaining plasma homocysteine levels and an adequate level of DNA methylation, a result we attribute to endogenous folate content. PMID- 9742466 TI - Increased fecal frequency and gastrointestinal symptoms following ingestion of galacto-oligosaccharide-containing yogurt. AB - Gastrointestinal symptoms and fecal frequency following ingestion of yogurt containing 15 g of galacto-oligosaccharides (GOS) per d were observed in 12 healthy volunteers. The effect of GOS on intestinal microflora was also studied in six volunteers. Defecation frequency increased during the administration period, but gastrointestinal symptoms, especially flatulence, also increased. The level of fecal bifidobacteria did not increase by the yogurt intake, but a significant increase was observed in the fecal bacteria growing on MRS media. The results indicate that dietary GOS increase gastrointestinal symptoms and fecal frequency in normal adults and have an effect on intestinal microecosystem. PMID- 9742467 TI - Effect of L-lactic acid on calcium absorption in rats fed omeprazole. AB - We examined the effect of L-lactic acid on calcium absorption in male Wistar rats made achlorhydric by dietary omeprazole, a proton pump inhibitor. The dietary omeprazole intake (0.03 g/100 g of diet) increased the gastric pH and decreased the apparent calcium absorption ratio. Dietary famotidine (0.03 g/100 g of diet), an H2-receptor antagonist, and lower doses of omeprazole (0.005 or 0.01 g/100 g of diet) did not affect the gastric pH or the calcium absorption. In a second experiment, dietary lactic acid (0.5, 1.0, or 2.5 g/100 g of diet) increased the intestinal calcium absorption dose dependently in rats fed omeprazole (0.03 g/100 g of diet). The gastric pH was significantly decreased only in the rats fed higher doses of lactic acid (1.0, or 2.5 g/100 g of diet). In a third experiment, a dietary sour milk beverage containing lactic acid (0.5 g/100 g of diet) increased the intestinal calcium absorption, but did not affect the gastric pH in rats fed omeprazole (0.03 g/100 g of diet). Although the significance of gastric acid in terms of overall calcium absorption is not known, under the present experimental conditions, the inhibition of gastric acid secretion by dietary omeprazole decreased the apparent calcium absorption, and the dietary lactic acid prevented the calcium absorption in rats fed omeprazole. PMID- 9742468 TI - The conservative radical. PMID- 9742469 TI - An analysis of posture and back pain in the first and third trimesters of pregnancy. AB - While the incidence of back pain during pregnancy has been shown to be high, few studies have investigated postural changes that occur during pregnancy and their relationship to back pain. The purpose of this study was to determine if posture and back pain changed from the first to the third trimester of pregnancy and whether there was a relationship between the two. Twelve healthy women who were having uncomplicated pregnancies participated in the study. During the first and third trimesters, each subject had their standing posture and back pain assessed by a Metrecom Skeletal Analysis System and a 0- to 10-cm line pain scale, respectively. Repeated measures analysis of variance and Pearson correlation coefficients were calculated on or between back pain and nine posture variables and revealed significant increases in third trimester back pain and postures compared with first trimester back pain (p < .05) and postures for lumbar angle (p < .01), posterior head position (p < .01), right pelvic sagittal tilt (p < .01), and left pelvic sagittal tilt (p < .01). No significant relationships were found between magnitude of or change in posture and back pain. These results suggest that in the standing position the lumbar lordosis and sagittal pelvic tilt increased and head position become more posterior as women progressed from the first trimester to the last trimester of pregnancy. These postural changes, however, were not related to back pain. This suggests that many of the posture correcting clinical exercise regimens given to pregnant women need to be investigated. PMID- 9742470 TI - A "step-to" gait decreases pressures on the forefoot. AB - Physical therapists use various gait training strategies to reduce stress on the lower extremities, but we could find no description or evaluation of the step-to gait using a cane. The purpose of this study was to evaluate the effect of a step to gait pattern and a cane on peak plantar pressures on the forefoot and the heel. Ten healthy subjects were evaluated (five females, five males, mean age = 24.6 +/- 4.9 years). In addition, one subject with peripheral neuropathy was tested to determine if a patient could be trained to use the step-to walking pattern and show similar results. All subjects were instructed in four walking conditions; step-to with and without a cane and step-through with and without a cane. Walking speed during the step-through pattern (normal walking) was matched to the speed of the step-to pattern. For the 10 healthy subjects, peak plantar pressures and walking speed of each of the four conditions were compared using a 2 x 2 repeated measures analysis of variance. One factor was gait pattern and one factor was use of a cane. Peak plantar pressures decreased an average of 53% on the forefoot but increased an average of 14% on the heel when subjects walked using step-to gait compared with a step-through gait. There was no effect due to use of a cane or walking speed between the conditions. The patient with peripheral neuropathy demonstrated a similar pattern but greater magnitude of changes compared with the healthy subjects. The foot initiating the step-to pattern showed a reduction in peak plantar pressures on the forefoot, probably because the foot remained flat during stance phase and a large push-off was not required. The step-to pattern, however, results in a slower and less symmetrical gait. The use of a step-to gait may be beneficial for patient populations that need to reduce plantar pressures on the forefoot. PMID- 9742471 TI - An electromyographical analysis of the scapular stabilizing synergists during a push-up progression. AB - Current literature recommends incorporating push-up progressions into upper extremity rehabilitation for advanced training of the scapular stabilizers. No documentation exists to demonstrate changes in the level of muscle activation when push-up progressions are performed. The purpose of this study was to determine the effect of difficulty level for push-ups on electrical activity of the scapular stabilizing synergists. Sixteen subjects performed five repetitions for each of the three conditions in a push-up progression. Electromyographic data collected on the serratus anterior, upper trapezius, and lower trapezius revealed a statistically significant interaction effect between the serratus anterior and upper trapezius and push-up condition when the feet were elevated. No significant interaction was found between push-up condition and the lower trapezius. This study supports the clinical use of push-up progressions to facilitate activation of the serratus anterior and the upper trapezius during upper extremity rehabilitation. PMID- 9742472 TI - Influence of hip position and gender on active hip internal and external rotation. AB - A general lack of descriptive details exists for measurements of hip rotation range of motion. This study was designed to establish the influence of gender and hip flexion position on active range of motion of the hip in external and internal rotation. Sixty (39 females and 21 males) healthy college-age (21.8 +/- 1.7 years) subjects were studied. Hip rotation of the dominant leg of each subject was measured in the prone (hip near 0 degree of flexion) and seated (hip near 90 degrees of flexion) positions using a standard goniometer. Data were analyzed using an analysis of variance model. Pearson's r statistics were used to determine the degree of association between measurements of hip rotation made seated vs. prone. A statistically significant difference (p < 0.05) was found between mean hip external rotation (ER) measured seated (36 +/- 7 degrees) and mean hip ER measured prone (45 +/- 10 degrees). Conversely, mean hip internal rotation (IR) measured seated (33 +/- 7 degrees) was not statistically different than mean hip IR measured prone (36 +/- 9 degrees). Females had statistically more active hip internal and external rotation than males (p < 0.05). A moderate degree of association existed between measurements of hip ER taken in the prone vs. seated position (r = 0.57, p < 0.05). For IR, the degree of association between the two measurement positions was slightly higher (r = 0.72, p < 0.05). Unlike the amount of active hip internal rotation which showed little difference between measurements made prone vs. seated, our data indicate that measurement position had a significant effect on the amount of active range of motion of the hip in ER. These findings are clinically significant for they stress the importance of documenting measurement position. They also stress the need for representative norms to be established for each hip position and gender. PMID- 9742473 TI - Motor skill acquisition strategies for rehabilitation of low back pain. AB - Evidence supporting the early use of exercise for the treatment of low back pain continues to grow. We must keep in mind, however, that motor skill learning and exercise are not synonymous. If rehabilitation goals are limited to the improvement of physical parameters (ie., strength, flexibility, endurance), the opportunity to help patients improve the performance of functional activities will be missed. The motor learning literature suggests several strategies for facilitating the acquisition of a motor skill: transfer-appropriate processing, the contextual interference effect, and repetitive self-evaluation. These techniques will cognitively challenge patients, helping them gain skills more quickly and retain them longer. By incorporating these methods into the rehabilitation program, patients will better transfer what they have learned from the rehabilitation environment to their everyday functional activities. PMID- 9742474 TI - A ballet dancer with chronic hip pain due to a lesser trochanter bony avulsion: the challenge of a differential diagnosis. AB - Physical therapy assessment involves confirmation of a medical diagnosis. To help discuss this process, this case report is presented regarding a ballet dancer who experienced 6 years of chronic hip pain and dysfunction. Many diagnostic tests and surgical procedures were performed by various physicians in search of a diagnosis. Physical therapy assessments did not support the working diagnoses, and treatment given according to evaluation findings was not effective. Initial hip radiographs revealed a bony ossicle at the lesser trochanter, which was overlooked. Hip radiographs taken 5 years later revealed the same bony ossicle. Ultimately, surgical removal of the ossicle eliminated the hip pain, and the patient returned to full activity and dance again. With the attempt to confirm the patient's diagnosis, the physical therapy approach to problem solving is discussed. This case gives an example where it is important to question the physician's diagnosis when the physical therapy assessment and treatment response do not support it. it is also evident that an in-depth physical therapy assessment may be self-limiting if pathology has not been ruled out properly by the physician. PMID- 9742475 TI - Stanley Paris Award Lecture. Reflections on the history and future of orthopaedic physical therapy. AB - Carolyn Wadsworth, MS, PT, CHT, OCS, deliver the 1998 Paris Distinguished Service Award Lecture at the Combined Sections Meeting in Boston in February. Wadsworth is the fifth recipient of the Paris Award, which is the Orthopaedic Section's highest honor given to commemorate a member's exceptional and enduring service. The Paris Distinguished Service Award was established in 1990 and is named after Stanley V. Paris, PhD, PT, founder and first president of the Orthopaedic Section. Born in Dunedin, New Zealand, Paris immigrated to the U.S. in 1966. He developed physical therapy practices in Boston, Hamilton, Bermuda, and Atlanta; established the Institute of Graduate Health Sciences in Atlanta, GA; and is currently president of the University of St. Augustine, St. Augustine, FL. Paris is a strong advocate of assertive professional practice, clinical specialization, and strengthening leadership in physical therapy. He also champions wellness, exemplified by his personal achievements in sailing the Atlantic Ocean, swimming the English Channel, and completing the Ironman Triathlon. Carolyn Wadsworth, recipient of the 1998 Paris Award, has served as secretary and president of the Orthopaedic Section and is currently editor of the Orthopaedic Section's Home Study Course. She owns a private practice, teaches nationally, and has written two books, Examination and Mobilization of the Spine and Extremities (1988) and Orthopedic Review for Physical Therapists (1998). Major components of the speech she presented at the Orthopaedic Section Awards Ceremony are highlighted in this article. PMID- 9742476 TI - Bridging the gap: reflections on the 12th World Aids Conference. PMID- 9742477 TI - Revision of HIV center medical staging scale. AB - HIV-related symptomatology represents both an indirect measure of immune functioning and a clinically significant indicator of disease progression. The most widely used classification system for HIV-related symptomatology is the Center for Disease Control (CDC) system. Although the CDC scale has widely recognized clinical utility, it provides only nominal-level measurement, which may be problematic for both clinicians and researchers. This article describes the revision and validation of the HIV Center Medical Staging Scale (rHCMSS), and ordinal-level physical illness scale that provides a means of independently measuring the progression of HIV-disease symptomatology and immunological decline. Concurrent use of the rHCMSS, the CDC classification system, and T lymphocyte and viral load data will provide a more comprehensive indication of HIV-disease progression for clinical and research purposes. PMID- 9742478 TI - Improved gastrointestinal tolerance and palatability of didanosine in adults by the use of pediatric power formulation. AB - Administration of didanosine (ddI) as buffered tablets is complicated by its poor palatability. The tolerance and palatability of pediatric powder ddI in an adult population of subjects with HIV infection as compared with buffered ddI tablets was compared. Twenty HIV-infected, were enrolled in a randomized crossover trial. Subjects took one of the two randomly assigned formulations for 4 weeks and then were crossed over to the alternative treatment. Twenty subjects were enrolled, and all of them completed the study. Of the 16 subjects, 10 reported that taking the ddI tablets affected the quality of their lives negatively. Of the participants, 18 (905) rated the pediatric formulation as 6 (better) or 2 (much better) than the buffered tablets. Preference was based on better taste and ease to swallow, although none of these 18 patients reported difficulties taking the pediatric formulation. In contrast, 19 of the 20 subjects reported difficulties taking the buffered tablets during the study period, including poor taste, trouble swallowing them, and diarrhea. Tolerance and palatability of ddI demonstrate marked improvement by the use of the pediatric powder, which suggests that the administration of ddI can markedly enhanced by the use of pediatric formulation. PMID- 9742479 TI - Triazole therapy for mucocutaneous candidiasis in HIV-infected patients. AB - Mucocutaneous candidiasis, such as oropharyngeal candidiasis, esophageal candidiasis, and vulvovaginal candidiasis, are common problems in patients with HIV infection. These conditions adversely affect patient quality of life and morbidity status. New oral triazole agents provide improved treatment options for patients with these and other opportunistic fungal infections; however, the development of resistance in some Candida species poses new challenges. This article provides an overview of the diagnosis of mucocutaneous candidiasis, current treatment modalities, concomitant drug interactions, common adverse drug reactions, and the emergence of fungal resistance, and it suggests nursing interventions to maximize patient benefits from antifungal therapy. PMID- 9742480 TI - Factors associated with vaginal yeast infections in HIV-positive women. AB - To better understand factors associated with symptomatic and asymptomatic vulvovaginal candidiasis, including the role of immune compromise and patient self-report, a cross-sectional analysis of factors associated with the isolation of yeast from vaginal swabs and clinical diagnosis of Candida vaginitis (CV) among 184 HIV-infected women was conducted. Sixty-four (35%) of the women had vaginal swabs positive for yeast. Nineteen (10%) women met the case definition for CV. In a logistic regression model, only CD4 < or = 100 cells/mm3 was predictive of CV (adds ratio = 4.5; 95% confidence interval = 1.0, 20; p = .05). The predictive value of patient self-report of CV was only 12%. This study demonstrates that all HIV-infected women should receive a regular and thorough gynecologic evaluation, regardless of self-reported symptoms. HIV-infected women will benefit from education about prevention and treatment of CV, and women whose CD4 counts are low may wish to consider prophylaxis for CV. PMID- 9742481 TI - Nursing practice for HIV/AIDS fever care: a descriptive study. AB - Nurses often perceive fever in relation to the impact it has on specific patient populations. Fever in HIV/AIDS patients is a common symptom. This descriptive study explored how nurses in AIDS care defined and described fever in HIV/AIDS patients and used nursing interventions for fever management. Seventy-five registered nurses, nurse practitioners, and LPNs employed in AIDS care throughout Florida were surveyed concerning interventions for AIDS-related fever. The study revealed that nurses in AIDS care treat fever by providing patient comfort, initiating nursing interventions for low-grade to moderate fever, and notifying the primary care provider if nursing interventions do not keep the temperature below 100.9 degrees F. The sample provided a variety of nursing interventions to make the patient more comfortable. In addition, the study also revealed a unique perspective for fever care, which influences the practice of nurses in AIDS care and might differentiate nursing practice in AIDS care from other nursing specialties. However, some interventions differed from fever nursing management strategies published by the Association of Nurses in AIDS Care. Further research needs to explore the efficacy of these interventions. PMID- 9742482 TI - Practice what we preach? HIV knowledge, beliefs, and behaviors of adolescents and adolescent peer educators. AB - The purpose of this article is to (a) describe the knowledge, beliefs, and sexual behaviors of urban adolescents and adolescent peer educators, and (b) identify elements needed to design effective HIV/AIDS prevention programs for out-of school youth. Thirty-three predominantly African American adolescents (female = 14; male = 19) between the ages of 14 and 24 in a large urban city including adolescent (n = 18) and adolescent peer educators (n = 15) participated. Paper and-pencil questionnaire and focus-group interviewing methods were used. Adolescents and adolescent peer educators had a moderately high level of HIV knowledge, confidence in their ability to use condoms, and beliefs that condom use would not decrease sexual pleasure or imply infidelity. Both groups reported low perceptions of susceptibility of HIV infection. Engagement in sexual risk behavior was low, but was significantly higher among males. Although adolescent male peer educators engaged in a higher frequency of risk behaviors over time, they had a lower frequency of sexual risk behaviors in the past 2 months compared with male adolescents. Study findings showed that HIV prevention interventions need to include information about specific risk behaviors, such as using condoms for oral sex, and cleaning drug paraphernalia. Community-based and church programs, visible HIV prevention messages, specifically those aimed at increasing perceptions of HIV risk, and the development of condom-use skills were identified by adolescents and adolescent peer educators as relevant approaches to reduce HIV infection among this population. PMID- 9742483 TI - HIV/AIDS in migrant and seasonal farm workers. PMID- 9742484 TI - Clinicians' antiretroviral medications guide. PMID- 9742485 TI - Reviewing the literature. PMID- 9742486 TI - Group sex in gay men: its meaning and HIV prevention implications. PMID- 9742487 TI - Cholestatic liver disease: an overview. PMID- 9742488 TI - Liver transplantation for cholestatic liver disease: screening and assessment of risk factors. AB - Orthotopic liver transplantation for primary biliary cirrhosis and primary sclerosing cholangitis is a well-accepted therapy for complications of end-stage liver disease and is associated with an excellent outcome in the majority of cases. However, transplant centers are striving to improve on these outcomes by studying ways to optimize the timing of transplantation. Several natural history and prognostic models for both primary biliary cirrhosis and primary sclerosing cholangitis have been derived from the study of large populations of patients in an attempt to predict long-term rates of survival. In addition, models exist to predict resource utilization after liver transplantation. Other factors besides complications of end-stage liver disease may also be indications for transplantation, including refractory pruritus, recurrent bacterial cholangitis in patients with primary sclerosing cholangitis, hepatic osteodystrophy, and a poor quality of life. PMID- 9742490 TI - Transplantation for childhood liver disease: an overview. AB - The advances made in surgical technique, postoperative care, and immunosuppression during the 1980s have permitted orthotopic liver transplantation (OLT) to evolve into an effective and widely accepted therapy for infants and children with end-stage liver disease. Biliary atresia, a progressive, obliterative disease of the bile ducts, is the most common indication for OLT in children, accounting for approximately 50% of cases. Metabolic liver disease (MLD) accounts for 20% to 25%; other common indications for OLT include fulminant hepatic failure (FHF) and forms of intrahepatic cholestasis. The principal problem associated with the increasing application of OLT is the burden placed on resources, particularly the availability of donor organs. The limited pool of size-matched donor organs has led to the application of a variety of alternatives to address the needs of the pediatric recipient; these include (1) reduced-size liver transplantation, (2) "split-liver" transplantation, and (3) use of living-related organ donors. In view of the impact on overall organ availability, the use of nontransplant options, including liver cell transplantation, especially for FHF and MLD, deserves broader application. Despite the success of transplantation, major challenges in childhood liver transplantation remain, including (1) improved preoperative management to ensure adequate growth, (2) more precise posttransplant management of immunosuppression to ensure graft viability and avoidance of lymphoproliferative disease, (3) earlier recognition of cytomegalovirus and Epstein-Barr virus infection, and (4) provision of services in a more cost effective manner. The ultimate solution is to prevent liver disease through vaccination and research. PMID- 9742489 TI - Liver transplantation for primary biliary cirrhosis and primary sclerosing cholangitis: does medical treatment alter timing and selection? AB - Liver transplantation is a highly effective treatment for patients with advanced primary biliary cirrhosis and primary sclerosing cholangitis. Transplantation is indicated when the patient's survival with transplantation is better than without or, earlier than this, if the patient's quality of life is intolerable from intractable fatigue or pruritus. Medical therapies for chronic cholestatic liver diseases are very limited. Ursodeoxycholic acid therapy in primary biliary cirrhosis reduces cholestasis and prolongs transplant-free survival; no other drugs are of proven efficacy in primary biliary cirrhosis, and none have any benefit on the disease progression of primary sclerosing cholangitis. Aggressive endoscopic therapy may produce symptomatic and biochemical improvement in primary sclerosing cholangitis but should be done without the expectation of retarding disease progression. Bilirubin is one of five criteria of the Child-Turcotte-Pugh score, which is necessary for the United Network for Organ Sharing listing for orthotopic liver transplantation. In addition, it is a major prognostic indicator in all the predictive models for primary biliary cirrhosis. Bilirubin reduction with ursodeoxycholic acid therapy in primary biliary cirrhosis appears to parallel disease severity, and prognostic models utilizing bilirubin retain their predictive power for survival even in treated patients. In summary, medical therapies for chronic cholestatic liver disease have very little effect on disease progression and, subsequently, on the timing or selection for transplantation. Liver transplantation is the only definitive therapy for primary biliary cirrhosis and primary sclerosing cholangitis. PMID- 9742491 TI - Biliary atresia--surgical management and treatment options as they relate to outcome. AB - Results show that the use of sequential surgical treatment, employing Kasai portoenterostomy in infancy, followed by selective liver transplantation for children with progressive hepatic deterioration yields improved overall survival. All children with successful Kasai portoenterostomy procedures who do not require OLT are survivors. Using newer transplant techniques, the 5-year survival rate for children who receive transplants with a primary diagnosis of biliary atresia was 82%. This yields an overall survival rate of 86% in this entire study population. Limited donor availability and increased complications after liver transplantation in infants less than 1 year of age mitigate against the use of primary liver transplantation without prior portoenterostomy for infants with biliary atresia. At present, these two operative procedures should be used as sequential and complementary modes of treatment rather than as competitive procedures. When biliary atresia is not recognized in infancy and established cirrhosis has resulted, primary transplantation should be offered as the initial surgical treatment. PMID- 9742492 TI - The liver and metabolic diseases of childhood. PMID- 9742494 TI - One center's experience with liver transplantation: alcohol use relapse over the long-term. PMID- 9742493 TI - Transplantation for alcoholic liver disease: a perspective from Europe. AB - It is now accepted that patients who receive a liver transplant for alcohol related liver disease have a rate of survival similar to those who receive grafts for other indications. Abstinence from alcohol before liver transplantation is important in ensuring that the liver will not recover, but the period of abstinence required before transplantation is undertaken is uncertain. Prognostic models for assessing patients with alcoholic liver disease have been developed but correlate poorly with each other. A return to alcohol consumption after transplantation is not uncommon, although graft failure or damage is uncommon. However, alcohol-related liver disease is becoming an increasing indication for liver transplantation. As the number of potential candidates exceeds the supply of donors, some form of rationing will be required. The general public places a lower priority on transplantation for alcoholic liver disease than for other indications, and this will need to be considered by those who allocate the donor livers. PMID- 9742495 TI - Hepatitis B and C: an overview. PMID- 9742497 TI - Management of posttransplantation viral hepatitis--hepatitis B. PMID- 9742496 TI - Hepatitis after transplantation. PMID- 9742498 TI - Management of posttransplantation viral hepatitis C. PMID- 9742499 TI - Hepatocellular carcinoma: risk factors and natural history. PMID- 9742500 TI - Treatment of hepatocellular carcinoma: medical options. AB - Nonsurgical therapy provides some benefit to patients with advanced hepatocellular carcinoma, although surgical options, including transplantation, remain the only chance for cure. Careful patient selection is required; patients with small nodular tumors may be considered for PEI therapy, whereas patients with larger tumors may be considered for TACE. Regardless of the treatment modality, the likelihood of survival is usually directly associated with the degree of hepatic dysfunction. Randomized, controlled trials of these treatment modalities are limited in number and design; therefore, it is difficult to assess their true impact on patient survival and quality of life. Secondary chemoprophylaxis against recurrent disease with vitamin A analogues is a promising adjunctive measure to both surgical and nonsurgical treatments for hepatocellular carcinoma. PMID- 9742502 TI - Shunt versus transplantation. PMID- 9742501 TI - Surgical options for hepatocellular carcinoma: resection and transplantation. AB - Surgical resection remains the best option for potential cure and long-term survival in patients with HCC. The question of to what extent transplantation for HCC should be performed remains controversial. There appears to be a definite role for OLT in the treatment of HCC, with many series showing improved survival over resection, especially with "favorable" tumors. What remains to be determined are the best patients and the best protocol. There is little question that patients with small unifocal tumors do well after OLT. It is the patient who falls outside of these narrow guidelines that poses a problem in clinical decision making and organ allocation. The ability to determine relative risk of recurrence of HCC would perhaps allow a more equitable allocation of a scarce resource. Currently, we evaluate each patient with HCC on an individual basis, making the best decision possible based on the patient's clinical status, our most advanced current imaging studies, and known clinical prognostic factors (Table 6). Adequate staging is essential to determine suitable candidates. Advances in multimodal adjuvant therapy are needed for patients with poor prognostic factors to achieve results similar to what is seen in those who receive transplants for nonmalignant diseases. Attempts at resection should be performed for those patients presenting with Child's class A cirrhosis, because these are the patients who would tolerate a resection with acceptable morbidity and mortality. Limited resections based on segmental anatomy may be consider in "good risk" Child's class B cirrhotics, considering the current organ shortage. Child's class C and decompensated Child's class B patients without significant risk factors should be evaluated for transplantation, and preoperative chemoembolization should be considered to prevent spread while the patient is on the waiting list. These patients should be monitored with imaging studies and by AFP levels on a regular basis while they await their transplant. After transplantation, chemotherapy should be considered for those patients with moderate to high risk of recurrence, within the guidelines of an institutional or multicenter protocol. In patients with multiple poor prognostic factors, or those who are too ill to undergo resection or transplantation, palliative measures may be used. As the need for organs increases, and the wait continues to grow, it becomes increasingly difficult to justify the use of a scarce resource for patients with a known less desirable outcome. On the other hand, we must be careful not to exclude an entire group of patients from a potentially curative procedure. We now have evidence that survival after transplantation for HCC in carefully chosen patients can equal that of benign disease. We need to be selective and cautious in our choice of recipients, but not exclusive, using prior experience and the knowledge we now possess regarding a set of fairly well delineated risk factors. PMID- 9742503 TI - Summary of guidelines on organ allocation and patient listing for liver transplantation. AB - In summary, a Child-Pugh class A cirrhotic patient without decompensation has a relatively long and stable natural history, but poor survival once decompensation with ascites or variceal bleeding has occurred. Thus, reasonable criteria for listing a patient for liver transplantation should be clinical decompensation of cirrhosis, particularly ascites or variceal bleeding, or combined clinical decompensation and biochemical deterioration of hepatic synthetic function that meets criteria for Child-Pugh class B or C status. On the basis of analysis of data regarding the natural history of compensated and decompensated cirrhosis, general principles of a minimal listing criteria were proposed (Table 5). These principles were used to establish non-disease-specific minimal listing criteria that are broadly applicable to all types of chronic liver diseases (Table 6). Some examples of proposed disease-specific minimal listing criteria are shown in Table 7. PMID- 9742504 TI - Matching donors and recipients. AB - This study identifies the major risk factors associated with outcome after liver transplantation, showing that candidates for this surgery can be stratified into differential risk categories at the time of the actual surgery. All the livers used were flushed with University of Wisconsin solution. The study is a retrospective multivariate analysis of 2376 consecutive transplantations performed on 2019 recipients between November 1, 1987, and December 31, 1993. Donor variables studied were age, sex, blood type, cause of death, intensive care unit length of stay, body mass index, use of pressors (dopamine infusion > 10 micrograms/kg/min or continuous infusion of epinephrine or norepinephrine), use of pitressin, cardiopulmonary resuscitation, terminal transaminase levels, serum sodium level at procurement, and total ischemia time. Recipient variables studied were age; sex; blood type; indication for liver transplantation; history of liver transplantation or upper abdominal surgery; United Network for Organ Sharing urgency status; need for mechanical ventilation; primary immunosuppression; and preoperative bilirubin level, prothrombin time, and creatinine level. The variables independently associated with outcome were donor age, female donor sex, ischemia time, recipient age, prior liver transplant, preoperative mechanical ventilation, preoperative bilirubin level, preoperative creatine level, indication for transplantation and primary immunosuppression used. The results of this study not only give us insight into the probable outcomes of individual patients, but also show that this stratification can be useful when comparing results across different groups or in helping to choose the best donor-recipient combination based on the calculated probability of a favorable outcome. PMID- 9742505 TI - Beta-cell apoptosis in an accelerated model of autoimmune diabetes. AB - BACKGROUND: The non-obese diabetic (NOD) mouse is a model of human type 1 diabetes in which autoreactive T cells mediate destruction of pancreatic islet beta cells. Although known to be triggered by cytotoxic T cells, apoptosis has not been unequivocally localized to beta cells in spontaneously diabetic NOD mice. We created a model of accelerated beta-cell destruction mediated by T cells from spontaneously diabetic NOD mice to facilitate the direct detection of apoptosis in beta cells. MATERIALS AND METHODS: NOD.scid (severe combined immunodeficiency) mice were crossed with bm1 mice transgenically expressing the costimulatory molecule B7-1 (CD80) in their beta cells, to generate B7-1 NOD.scid mice. Apoptosis in islet cells was measured as DNA strand breakage by the TdT mediated-dUTP-nick end labeling (TUNEL) technique. RESULTS: Adoptive transfer of splenocytes from spontaneously diabetic NOD mice into B7-1 NOD.scid mice caused diabetes in recipients within 12-16 days. Mononuclear cell infiltration and apoptosis were significantly greater in the islets of B7-1 NOD.scid mice than in nontransgenic NOD.scid mice. Dual immunolabeling for TUNEL and either B-7 or insulin, or the T cell markers CD4 and CD8, and colocalization by confocal microscopy clearly demonstrated apoptosis in beta cells as well in a relatively larger number of infiltrating T cells. The clearance time of apoptotic beta cells was estimated to be less than 6 min. CONCLUSIONS: B7-1 transgenic beta cells undergo apoptosis during their accelerated destruction in response to NOD mouse effector T cells. Rapid clearance implies that beta cells undergoing apoptosis would be detected only rarely during more protracted disease in spontaneously diabetic NOD mice. PMID- 9742506 TI - The 12 kD FK 506 binding protein FKBP12 is released in the male reproductive tract and stimulates sperm motility. AB - BACKGROUND: The 12 kD FK506 binding protein FKBP12 is a cytosolic receptor for the immunosuppressant drugs FK506 and rapamycin. In addition to its critical role in drug-induced T-cell immunosuppression, FKBP12 associates physiologically with ryanodine and inositol 1,4,5-trisphosphate (IP3) receptors, regulating their ability to flux calcium. We investigated a role for FKBP12 in male reproductive physiology on the basis of our identification of extremely high levels of [3H]FK506 binding in male reproductive tissues. MATERIALS AND METHODS: [3H]FK506 binding studies were performed to identify tissues enriched with FK506 binding sites. The abundant [3H]FK506 binding sites identified in the male reproductive tract were localized by [3H]FK506 autoradiography. FK506 affinity chromatography was employed to purify FKBP from epididymal fluid. Anti-FKBP12 Western analysis was used to confirm the identity of the purified FKBP. The binding of exogenous FKBP12 to sperm was evaluated by [32P]FKBP12 binding studies and [33P]FKBP12 autoradiography. The effect of recombinant FKBP12 on sperm motility was investigated using a Hamilton Thorne motility analyzer. RESULTS: Male reproductive tissues contained high levels of [3H]FK506 binding. The localization of [3H]FK506 binding sites to the tubular epithelium of the caput epididymis and the lumen of the cauda and vas deferens suggested that FKBP is released in the male reproductive tract. FKBP12 was purified from epididymal plasma by FK506 affinity chromatography. Radiolabeled FKBP12 specifically bound to immature but not mature sperm. In sperm motility studies, FKBP12-treated caput sperm exhibited double the curvilinear velocity of untreated controls. CONCLUSIONS: High levels of FKBP12 are released in the male reproductive tract and specifically associate with maturing sperm. Recombinant FKBP12 enhances the curvilinear velocity of immature sperm, suggesting a role for FKBP12 in motility initiation. The highest concentrations of soluble FKBP12 in the male reproductive tract occur in the lumen of the vas deferens, a site of sperm storage and the conduit for ejaculated sperm. Preservation of mammalian sperm for reproductive technologies may be optimized by supplementing incubation or storage media with FKBP12. PMID- 9742507 TI - High concentration of L-arginine suppresses nitric oxide synthase activity and produces reactive oxygen species in NB9 human neuroblastoma cells. AB - Hereditary argininemia manifests as neurological disturbance and mental retardation, features not observed in other amino acidemias. The cytotoxic effect of a high concentration of L-arginine (L-Arg) was investigated using NB9 human neuroblastoma cells (NB9), which express neuronal nitric oxide synthase (nNOS). When the concentration of L-Arg in the medium increased from 50 microM to 2 mM after incubation for 48 hr, the intracellular concentration of L-Arg increased from 68.0 +/- 1 pmol/10(6) cells to 1310.0 +/- 5 pmol/10(6) cells and that of L citrulline (L-Cit) from undetectable levels to 47.1 +/- 0.2 pmol/10(6) cells (mean +/- SD of three independent analyses). This increase in intracellular L-Arg levels caused a decrease in NOS activity by approximately 71%. Flow cytometric analysis showed that reactive oxygen species (ROS) are produced in NB9 exposed to 2 mM L-Arg. The production of ROS was abolished by a NOS inhibitor, NG-nitro-L arginine-methylester. Production of ROS was also observed when NB9 were treated with L-Cit for 48 hr. To investigate the effect of L-Cit on the activity of NOS, a kinetic study on nNOS was conducted using cellular extracts from NB9. The apparent Km value of nNOS for L-Arg was 8.4 microM, with a Vmax value of 8.2 pmol/min/mg protein. L-Cit competitively inhibited NOS activity, as indicated by an apparent Ki value of 65 nM. These results suggest that L-Cit formed by nNOS in L-Arg-loaded neuronal cells inhibits NOS activity and nNOS in these L-Arg-loaded cells functions as a NADPH oxidase to produce ROS, which may cause neurotoxicity in argininemia. PMID- 9742509 TI - On the nature and pattern of neurocognitive function in major depressive disorder. AB - An effect size analysis of neurocognitive function in patients with major depressive disorder using meta-analytic principles was conducted. The results from 726 patients with depression and 795 healthy normal controls revealed that depression had the largest effect on measures of encoding and retrieval from episodic memory. Intermediate effect sizes were recorded on tests of psychomotor speed and tests that require sustained attention. Minimal effect sizes were found on tests of semantic memory, primary memory, and working memory. Moreover, major depressive disorder is accompanied by dysfunction of effortful encoding of information along with an accompanying inefficiency of retrieving poorly encoded information from declarative memory. PMID- 9742510 TI - Cognitive deficits, psychopathology, and psychosocial functioning in bipolar mood disorder. AB - The objective of this study was to study the relationship of poor functioning, cognition, and psychopathology in bipolar mood disorder. The authors assessed 36 patients with bipolar mood disorder (23 VA, 13 community) for the presence of psychopathology, cognitive deficits, and psychosocial impairment. The authors assessed psychopathology using screening and follow-up questions based on the schedule for affective disorder and schizophrenia, lifetime version (SADS-L), schedule for the assessment for negative symptoms (SANS), and schedule for the assessment of positive symptoms (SAPS), and psychosensory features using the "Profile of Psychomotor Symptoms." They tested cognitive functioning in the following domains: 1) general intelligence and language, 2) verbal and visual memory, and 3) visuospatial functioning. They also assessed psychosocial functioning using a structured scale to assess maladjustment and an impairment rating scale. Patients with bipolar disorder showed significant impairment compared to age equivalent normals in several cognitive domains. Anhedonia was related to memory deficits. Memory deficits were also associated with poor psychosocial functioning. This study demonstrates that nondemented, asymptomatic patients with bipolar disorder exhibit substantial cognitive deficits that are associated with poor functioning, and anhedonia and avolition best predict this outcome. PMID- 9742508 TI - Frequent N addition and clonal relatedness among immunoglobulin lambda light chains expressed in rheumatoid arthritis synovia and PBL, and the influence of V lambda gene segment utilization on CDR3 length. AB - BACKGROUND: In rheumatoid arthritis (RA), B-lineage cells in the synovial membrane secrete large amounts of immunoglobulin that contribute to tissue destruction. The CDR3 of an immunoglobulin light chain is formed by rearrangements of VL and JL gene segments. Addition of non-germline-encoded (N) nucleotides at V(D)J joins by the enzyme terminal deoxynucleotidyl transferase (TdT) enhances antibody diversity. TdT was previously thought to be active in B cells only during heavy chain rearrangement, but we and others reported unexpectedly high levels of N addition in kappa light chains. We also found clonally related kappa chains bearing unusually long CDR3 intervals in RA synovium, suggesting oligoclonal expansion of a set of atypical B lymphocytes. In this study, we analyzed lambda light chain expression to determine if N addition occurs throughout immunoglobulin gene rearrangement and to compare CDR3 lengths of lambda and kappa light chains in RA patients and normal individuals. MATERIALS AND METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) amplification of V lambda III transcripts was performed on RA synovia and peripheral blood lymphocytes (PBL) and normal PBL for which kappa repertoires were previously analyzed. Representative lambda + PCR products were cloned and sequenced. RESULTS: Analysis of 161 cDNA clones revealed that N addition occurs in lambda light chains of RA patients and normal controls. The lambda light chain repertoires in RA were enriched for long CDR3 intervals. In both RA and controls, CDR3 lengths were strongly influenced by which V lambda gene segment was present in the rearrangement. Five sets of clonally related sequences were found in RA synovia and PBL; one set was found in normal PBL. CONCLUSIONS: In humans, unlike mice, N addition enhances antibody diversity at all stages of immunoglobulin assembly, and the structural diversity of lambda CDR3 intervals is greater than that of kappa light chains. Clonally related V lambda gene segments in RA support an antigen-driven B-cell response. PMID- 9742511 TI - Neurocognitive deficit in fronto-temporal dementia. AB - Meta-analytic principles were used to formulate a neurocognitive profile of fronto-temporal dementia (FTD). The neurocognitive test results from a total of 88 patients with FTD and 100 health controls were synthesized using effect size analyses. The results indicate that patients with FTD are most deficient on tests of cognitive flexibility and abstraction, followed by performance on measures of performance skill, memory acquisition, attention/concentration, delayed recall, and, finally, verbal skill. A rank-order profile of specific neurocognitive tasks and test variables in order of sensitivity of FTD is also provided to help interpret the quantitative results. On the basis of the profile, neurocognitive markers meeting heuristic benchmark criteria were identified that can aid in the differentiation of FTD from other disorders with prominent features of dementia. PMID- 9742512 TI - Neuropsychological and motor functioning after unilateral anatomically guided posterior ventral pallidotomy. Preoperative performance and three-month follow up. AB - This study presents baseline and 3-month follow-up motor and neuropsychological data for 22 patients with Parkinson's disease (PD) who underwent anatomically guided unilateral posterior ventral pallidotomy (PVP). Postsurgical improvements were seen in psychomotor speed, fine motor accuracy, and dyskinesia, whereas grip strength decreased on the side contralateral to the surgery. No change was detected in overall level of cognitive functioning, nor were changes demonstrated in memory, language, or working memory when the entire sample of patients was evaluated. When the group was divided on the basis of side of surgery, patients with left-sided pallidotomies showed a decline in verbal fluency. Patients and caregivers reported improvement in psychosocial functioning. These initial findings of improved motor performance and largely unaffected cognitive functions are consistent with results obtained with functional PVP and provide support for the use of anatomically guided posterior ventral pallidotomy in the treatment of motor symptoms of PD. PMID- 9742513 TI - Predictors of involvement in P300 latency in solvent-exposed adults. AB - Persons with a history of exposure to organic solvents have been shown to have cognitive and personality changes, as well as abnormalities on measures of neurophysiology (e.g., delays in P300 latency). Studies assessing long-term sequelae in exposed persons have been limited, especially those using neurophysiologic measures. This study assessed cognitive event-related potentials (ERPs) in 16 persons with a history of organic solvent exposure at two testings, separated, on average, by 1.5 years. The sample was divided into persons who showed improvement on P300 latency (e.g., reduction in latency of 1.5 SD of control group) and those who did not. Sixty-three percent showed no improvement, whereas 37% showed significant improvement. Recency of exposure and the interaction of exposure duration and history of peak exposure significantly predicted group membership. That is, persons with shorter duration of exposure coupled with no peak exposures and longer time from exposure to test were more likely to fall in the improved group. Substituting age for duration of exposure in the interaction term improved classification of the two groups. The results support previous findings that most exposed persons do not show significant improvements over time. The results further suggest that there is a need to assess factors, such as aging, which may make one more vulnerable to the neurotoxic effects of solvents. PMID- 9742514 TI - Different psychological status in the two hemispheres of two split-brain patients. AB - Questions of a psychological nature were presented to two split-brain patients from the California series encouraging each hemisphere to respond simultaneously and independently. The responses of both patients indicated that their hemispheres were responding independently. For the first patient, his right hemisphere appeared to be more disturbed than his left by childhood memories of being bullied. The right hemisphere of the second patient seemed to regard himself more positively, but it also seemed to feel more negative emotions such as loneliness and sadness. We discuss the possible significance of the findings. PMID- 9742515 TI - Catatonia-like events after valproic acid with risperidone and sertraline. AB - A patient with schizoaffective disorder developed signs of catatonia while on a regimen of valproic acid, sertraline, and risperidone. The catatonic features evolved for the first time after a single dose of valproate and were alleviated by lorazepam. The same catatonic signs recurred after a second dose of valproate and again remitted after lorazepam. Catatonia did not recur subsequent to discontinuing valproate, and the patient had tolerated the combination of valproate and risperidone in the past without developing catatonia. Although the catatonia in this case initially appears to be paradoxical, the phenomenon is actually consistent with current models of catatonia. A unique drug interaction may account for this phenomenon. Possible pharmacokinetic and pharmacodynamic explanations are discussed, including the relation of catatonia to increased activity of the medial globus pallidus and current models of catatonia. PMID- 9742516 TI - Pseudoakathisia from a acquired lesion. AB - Akathisia, "inability to remain seated", is a common condition in patients medicated with neuroleptics, and also occurs in other medication regimens. Only a few cases of akathisia resulting from focal lesions have been published. These cases suggest that differing lesion sites are sufficient for producing objective or subjective akathisia. The authors present a patient with akathisia who had no subjective complaints, suggesting that a combined basal ganglia and frontal lobe lesion may lead to objective akathisia (pseudoakathisia). PMID- 9742517 TI - The concept of pseudoschizophrenia. PMID- 9742518 TI - [Dysfunction of E-cadherin-catenin system in invasion and metastasis of colorectal cancer]. AB - The E-cadherin-mediated cell adhesion system is now considered to be an "invasion suppressor system" in cancer cells. Dysfunction of the E-cadherin system due to mutations of the genes of E-cadherin and catenins has not been reported in colorectal cancer. Histologically, well-differentiated colorectal cancer cells are found to be scattered at the invasive front in primary lesions and form glands again in metastatic sites. We have reported the association and presence of signal transduction between c-erbB-2/epidermal growth factor receptor (EGF-R) and beta-catenin in human cancer cells. This temporal dysfunction of the E cadherin system observed in colon cancers may be caused by tyrosine phosphorylation of beta-catenin through activated receptor-type tyrosine kinases. Overexpression of EGF-R and tyrosine phosphorylation of beta-catenin are often observed in "focal dedifferentiated cells" at the invasive front of colorectal cancers. In addition, beta-catenin expression is regulated by the APC tumor suppressor gene product. Thus the E-cadherin-catenin system may play important roles not only in invasion and metastasis but also in the carcinogenesis of colorectal cancer. PMID- 9742519 TI - [Expression of variant CD44 in colorectal cancer and its relationship to liver metastasis]. AB - A number of different isoforms of CD44 generated by alternative splicing have been isolated and sequenced. There have been several reports that CD44v plays a role in the steps of the metastatic process. We examined the role of the variant CD44v8-10 in metastases of human colon cancer cell line HT29m using a monoclonal antibody reactive with the v9 product (mAb 44-1V). Pretreatment with mAb 44-1V prevented the formation of liver metastases. In addition, we found that the attachment of HT29m cells to the basement membrane matrix was inhibited by mAb 44 1V. Several reports have shown correlations between metastatic potential and expression of CD44v in human colorectal cancer. We demonstrated that CD44v8-10 and CD44v6 RNA expression was higher in carcinomas associated with liver metastases than in those without by Northern blotting. We analyzed the expression of the CD44v8-10 product in colorectal cancer immunohistochemically using mAb 44 1V, and evaluated its prognostic significance. There were significant correlations between CD 44v8-10 immunoreactivity and both lymph node and liver metastases. Patients with CD44v8-10-positive tumors had a greater relative risk of death compared with those whose tumors were CD44v8-10 negative. These results suggest that CD44v8-10 may play an important role in the adhesion of tumor cells to the capillaries of distant organs in the metastatic process, and that immunohistochemical detection of CD44v8-10 may be a biologic marker of prognostic significance. PMID- 9742520 TI - [Regulation of integrin function in the metastasis of colorectal cancer]. AB - Alterations in several classes of adhesion molecule have been implicated in the progression of colorectal cancer. Cell adhesion regulator (CAR) has been identified as a regulator molecule of integrin-dependent cell adhesion. We have explored the possible involvement of the CAR gene in colorectal cancer. Reverse transcription-PCR revealed that CAR expression was detected in normal colonic cells, whereas it was decreased or undetectable in 6 of 13 (46.2%) human colon cancer cell lines. Adhesion of HT-29 cells to extracellular matrix components was up-regulated by the introduction of CAR. CAR-transfected HT-29 cells showed a significantly reduced spontaneous metastatic potential in nude mice. In 14 of 30 cases (46.7%), CAR expression in cancer was less than one-tenth of that in matched noncancerous tissue. The tumor: normal ratio of CAR expression was significantly lower in patients with lymph node metastases than in those without (p < 0.01) and in patients with distant metastases than in those without (p < 0.05). CAR expression was significantly lower in more advanced Dukes' stage tumors (p < 0.05). Our results suggest that down-regulation of CAR expression may play an important role in the progression and metastasis of colorectal cancer. PMID- 9742521 TI - [Carbohydrate chains and hematogenous metastasis: a review]. AB - It is well known that the expression of carbohydrate chains on cell changes during the development and progression of carcinoma. Some carbohydrate chains have been used in clinical practice as tumor markers. Some, such as sialyl Lewis a (CA 19.9), sialyl Lewis X, and Tn, are known to function as adhesion molecules. However, their role in the development of hematogenous metastasis is still controversial. In this paper, many investigations concerning this issue are reviewed. Clinical trials for treatment of patients with advanced cancer using carbohydrate chains are also mentioned. PMID- 9742522 TI - [Involvement of cytokines in invasion and metastasis of colon cancer]. AB - Despite improvement in early diagnosis, surgical techniques, and general patient care, most deaths of cancer patients result from metastases. Recent studies have revealed that cytokines produced by cancer cells or by stromal cells play an important role into development the cancer metastasis. The formation of a cancer metastasis involves several major steps: 1) extensive vascularization; 2) local invasion; 3) adherence either to capillary endothelial cells or to subendothelial basement membrane; 4) extravasation; and 5) proliferation. In colon cancer, several cytokines such as growth factors, inflammatory cytokines, and angiogenic factors have been confirmed to be involved in each step of metastasis. This paper summarizes the involvement of cytokines in the development of invasion and metastasis in colon cancer. PMID- 9742523 TI - [Multidisciplinary treatment for colorectal liver metastases]. AB - The liver is an large immunologic organ with liver-associated macrophages (Kupffer cells) and natural killer-like primitive T cells. As these effectors are activated by interleukin-2 (IL-2), we have administered IL-2-based hepatic arterial infusion therapy in the treatment of patients with liver metastases of colorectal cancer. Patients with unresectable liver metastases were administered IL-2 7 x 10(5) U and 5-fluorouracil (5-FU) 250 mg/day as a continuous infusion, with a bolus injection of mitomycin C (MMC) 4 mg once weekly. Of 25 patients treated with this regimen, 19 achieved complete or partial responses (response rate: 76%). A multi-institutional randomized trial following the pilot study showed reproducible favorable results. For patients with resectable metastases, we have administered this infusion therapy for the prevention of cancer recurrence in the liver. Patients who had undergone curative hepatectomy received IL-2 1.4 to 2.1 x 10(6) U, 5-FU 250 mg and MMC 2 to 4 mg weekly for 6 months. Of 18 patients, 12 are alive disease-free, and the 5-year overall survival rate is 75%. Recurrent cancer has developed in 6 of the 18 patients; however, no patients had recurrence in the residual liver. We believe that liver metastases of colorectal can be controlled by this multimodal treatment. PMID- 9742524 TI - [Implication of VEGF and MMPs in hepatic metastasis of human colon cancer]. AB - This article describes the significance of mRNA expression of VEGF, MMP-2, MMP-9, and MT1-MMP in human colorectal cancer metastases, particularly hepatic metastases. The levels of gene expression were quantified by Northern blot hybridization in tumor and nontumor tissues obtained from 66 primary cases. Significantly higher levels of expression of VEGF mRNA were observed in patients with synchronous hepatic metastases (n = 15) and/or lymph node metastases than in those without. Patients with synchronous hepatic metastases had significantly higher levels of mRNA expression of all MMP genes than in those without, and no apparent correlation was seen between MMP mRNA expression and other clinicopathologic variables. Also in a study including 4 cases of metachronous hepatic metastases after surgery. VEGF, MMP-9, and MT1-MMP mRNA expression were significantly higher in patients with hepatic metastases than in those without, indicating that these are predictable markers for hepatic metastases. Immunohistochemical examination revealed that VEGF and MT1-MMP were localized mainly in cancer cells, whereas MMP-2 and MMP-9 were distributed throughout stromal cells such as fibroblasts and leukocytes in tumor tissues. PMID- 9742525 TI - [Therapeutic effect of angiogenesis inhibitors on liver metastases of human colorectal carcinoma]. AB - Angiogenesis is essential for the growth of solid tumors. Antiangiogenic therapy has therefore attracted considerable interest as a novel therapy for various tumors including colorectal carcinoma. We experimentally investigated the therapeutic effect of TNP-470, a nonspecific inhibitor of angiogenic factors, and vascular endothelial growth factor (VEGF)-neutralizing antibody (VEGFAb), was a VEGF-specific inhibitor, on liver metastases of colon carcinoma using a murine orthotopic transplantation model. TK-4 and TK-13 are moderately differentiated adenocarcinoma strains established in our department which express VEGF mRNA and protein. Administration of TNP-470 30 mg/kg significantly inhibited the liver metastases of both strains, as did administration of VEGFAb 100 micrograms/mouse. The therapeutic effect on liver metastases was more dominant with antiangiogenic therapy than with chemotherapy (mitomycin C). Furthermore, the sustained effect of TNP-470 induced tumor dormancy and consequently improved the survival of the animals. These results suggest that antiangiogenic treatment will be a potent therapy for liver metastases of human colorectal carcinoma in the future. PMID- 9742527 TI - [Host reactions affect cancer progression]. AB - The tumor-host relationship is one of the factors determining the biological behavior of malignant tumors. In colon cancer, the costimulatory molecules B7 1/B7-2 are expressed by macrophages distributed along the invasive margin (tumor host interface). T-lymphocytes are also distributed in the same area with direct cell-to-cell contact with these B7+ macrophages. The distribution of these macrophages is lower in colon cancer patients with liver metastasis. CD8+ T cells are distributed within cancer cell nests of colon cancer. The survival rate of patients with higher levels of these T-cells is favorable. Our data suggest the presence of a host immune reaction by macrophages and T-lymphocytes, which diminishes the aggressiveness of cancer cells. As shown in the present paper, comparative analysis is required to assess the biological behavior of cancer. PMID- 9742526 TI - [PyNPase expression and cancer progression in the colorectum]. AB - We analyzed PyNPase expression discriminating between cancer and tumor stroma of the colorectum by Western blotting using a newly developed extraction method from microdissected tissue sections fixed with buffered formalin. Analysis of 98 colorectal cancers revealed that PyNPase was as high as 70.2 +/- 18.5 unit/mg protein in the stroma fraction (SF), whereas it was 45.1 +/- 10.5 in the cancer fraction (CF) (p < 0.0001). Vessel density was correlated with PyNPase in the SF but not in the CF. In stage IIIb, 11 cases expressing a high level of PyNPase in the CF showed poorer prognosis than 10 cases with low-level PyNPase expression (p < 0.05), although the level of PyNPase expression in the SF did not affected the patients prognosis. Immunohistochemical examination indicated that PyNPase in the SF was mainly produced by macrophages (M phi), and therefore we investigated the profile of PyNPase production by M phi. In in vitro experiments PyNPase production by M phi was greatly enhanced by stimulation with OK-432, and the culture supernatant had the ability to convert 5'DFUR to 5-FU. PMID- 9742528 TI - [Messenger RNA expression of p21WAF1/CIP1 in colorectal carcinoma tissues]. AB - p21WAF1/CIP1 is a potent, tight-binding inhibitor of Cdks which is induced by the wild-type p53 gene. We examined p21WAF1/CIP1 mRNA expression in 16 surgically excised human colorectal tumor and nontumor tissues using Northern blot analysis with reference to the identification of p53 gene mutation. p53 gene mutation was detected in six tumor tissues but not in the other 10 using the PCR-SSCP method. The p21WAF1/CIP1 mRNA level was lower in tumor tissues than in the corresponding nontumor tissues. The mean expression level of p21WAF1/CIP1 mRNA was lower in tissues in which the p53 gene mutation was detected than in those in which it was not, although the difference was not significant. The relative p21WAF1/CIP1 mRNA expression level was significantly higher in the tumors with liver metastases than in those without. These results suggest that the p21WAF1/CIP1 mRNA expression level, which might be a indicator of colorectal cancer malignancy, is suppressed in human colorectal tumor tissues compared with the corresponding nontumor tissues and that the wild-type p53 gene is one of the factors inducing p21WAF1/CIP1 mRNA expression. PMID- 9742529 TI - [Molecular surgery for human colorectal cancer with tumor suppressor p53 gene transfer]. AB - Recent advances in molecular biology have demonstrated that multistep genetic alterations are involved in the carcinogenesis of human colorectal cancer and that alteration of the p53 gene by mutation, deletion, or rearrangement is a major factor in this process. Human gene therapy has become a reality with the development of effective techniques for delivering the gene to the target cells. The efficacy of gene therapy for various types of genetic disease now being evaluated in clinical trials. These findings led us to develop a novel gene therapeutic strategy for human colorectal cancer that could replace the abnormal p53 gene using a recombinant, replication-defective adenoviral vector (termed Adp53). Infection with Adp53 induced rapid apoptotic cell death in DLD-1 and LoVo human colorectal cancer cell lines differing in their p53 status. Treatment with cisplatin following infection with Adp53 significantly suppressed the growth of WiDr colorectal cancer cells compared to single treatments alone. Thus restoration of wild-type p53 function exhibited an antitumor effect by inducing apoptosis as well as by markedly enhancing the effect of common chemotherapeutic agents in human colorectal cancer cells. In addition, Adp53 infection was antiangiogenic in SW620 human colorectal cancer cells. The application of this technology to human cancer therapy is now in progress. The article reviews recent highlights in this rapidly evolving field. PMID- 9742530 TI - Paediatric patients: SIDS, safety and positioning. PMID- 9742531 TI - Intraoperative ventilation. PMID- 9742532 TI - Distress at induction of anaesthesia in children. A survey of incidence, associated factors and recovery characteristics. AB - This study analysed the frequency of distress at induction (DAI) in 2122 paediatric patients. The data were analysed to assess predictors of DAI and to examine associations between predictors of DAI and recovery characteristics. Patient age, preoperative behaviour, premedication (oral midazolam, n = 480) and venue for anaesthesia induction were associated with changes in the incidence of DAI. Distressed preoperative behaviour was a good predictor of DAI in all age groups. Premedication reduced the incidence of DAI in children aged 0.5-2 years old, and in older children who were distressed preoperatively. Induction in the Day Surgery Unit was associated with a reduction of the incidence of DAI in younger children. Children with DAI were more likely to suffer from distress at arousal (P = 0.001). Average early recovery time was prolonged 4.4 minutes and average discharge time in day patients was delayed 36 minutes by the use of oral midazolam premedication. Premedication was not significantly associated with arousal distress. We conclude that a policy of optimizing nonpharmacological approaches for minimizing induction distress, combined with selective premedication with oral midazolam, can produce a low incidence of induction distress and adverse effects. PMID- 9742533 TI - Absence or presence of a leak around tracheal tube may not affect postoperative croup in children. AB - To prevent postoperative croup in children, many anaesthesiologists use a tracheal tube that allows a leak when tested with 20 to 25 cm of water pressure. We studied the correlation of postoperative croup with leak, duration of anaesthesia, and a recent cold in 159 healthy outpatient children who had strabismus correction by the same surgeon and the same anaesthesiologist. We found no correlation between the presence or absence of a leak and the incidence or severity of postoperative croup. There was a strong trend toward significance when postoperative croup and duration of anaesthesia were compared (P = 0.056) and a significant, positive correlation between severe croup (requiring racemic epinephrine) and duration of anaesthesia (P = 0.005). Patients having a recent cold did not have an increased incidence of postoperative croup. A leak around the tracheal tube at 20 to 25 cm of water pressure may not be required for a healthy child who undergoes surgery lasting less than 2 h if the child has no history of croup. PMID- 9742534 TI - Convective warming blankets improve peroperative heat preservation in congenital heart surgery. AB - Peroperative heat preservation, following hypothermic cardiopulmonary bypass (CPB) in children, has always been a challenge to the anaesthetist. We studied the efficiency of a convective heating system on peroperative heat preservation in 50 children undergoing congenital heart surgery. Twenty-five children, rewarmed by CPB and heating mattress, were randomly selected (Group 1). Another 25 children, rewarmed by CPB, heating mattress and convective warming blankets in addition (Group 2), were selected so the two groups were comparable regarding age, weight and anaesthetic management. The central and peripheral temperatures were measured during bypass, at the end of bypass and at the end of operation. A retrospective evaluation showed that during bypass the peripheral temperature was significantly lower in Group 2 than in Group 1, with no significant difference in central temperature. At the end of bypass there was no significant difference between the two groups. At the end of operation the central and peripheral temperatures were significantly higher in Group 2. In conclusion convective warming blankets are effective in keeping or even raising the temperature following congenital heart surgery. PMID- 9742535 TI - A comparison of three methods of analgesia in children having day case circumcision. AB - Sixty-one children were randomized to receive one of three methods of analgesia for day case circumcision. Group 1 received a penile block, group 2 received a penile block plus diclofenac suppository and group 3 received a diclofenac suppository alone. CHEOPS pain scoring was performed in the recovery area, one h after awakening and two h after awakening. There was no difference between the groups except in the recovery area when group 3 cried more and had a higher pain score than group 2. Parental follow-up questionnaires for the subsequent two days showed no difference in measured parameters between the groups. PMID- 9742536 TI - A timed reexpansion inspiratory manoeuvre (TRIM) for treating oxyhaemoglobin desaturation in children following a period of apnoea--studies in an animal model. AB - Our clinical experience has shown that the use of a constant distending airway pressure of 30 cm water for 10 s, termed a timed reexpansion inspiratory manoeuvre (TRIM), is often successful in correcting oxyhaemoglobin desaturation in anaesthetized children. The aim of this study was to assess the efficacy of TRIM in lambs. Following a standard relaxant anaesthetic, ventilation was stopped and oxyhaemoglobin saturation allowed to fall to 70% and the time taken to return to baseline was compared between three groups. The median time was 42.5 s when ventilation was restarted with 33% oxygen in nitrous oxide (33% group), 30 s when ventilation was restarted with 100% oxygen (100% group) and 22.5 s with a TRIM before restarting ventilation with 33% oxygen in nitrous oxide (TRIM group). The correction of desaturation was more rapid in the TRIM group compared with the 33% group (P < 0.004) and the 100% group (P < 0.003). Oxyhaemoglobin desaturation due to apnoea in anaesthetized lambs is more effectively treated with a TRIM than by increasing the inspired oxygen fraction. PMID- 9742537 TI - Oral transmucosal fentanyl citrate as an anaesthetic premedication when dosed to an opioid effect vs total opioid consumption. AB - Thirty min prior to anaesthetic induction for surgery, children aged 4-12 years old were given a 10 micrograms.kg-1 oral transmucosal fentanyl citrate (OTFC) and were instructed to suck the OTFC until pruritus appeared (Group 2) or until the entire dose was consumed (Group 1). Sedation, apprehension and cooperation scores were rated, and vital signs including oxygen saturation were monitored until anaesthetic induction. The results showed that pruritus was present in 76% of children; however; in all but one child, it occurred after the OTFC had been completely consumed. There were no significant changes in oxygen saturation, but respiratory rate decreased from 19.6 +/- 1.7 to 18.4 +/- 1.3. Activity decreased significantly; however, cooperation and apprehension did not change. The conclusion was that pruritus cannot be used as an endpoint for OTFC effectiveness; however, OTFC dosed at 10 micrograms.kg-1 is effective in providing sedation without causing clinically significant changes in vital signs or oxygen saturation. PMID- 9742538 TI - Haemodynamic improvement in a child with meningococcal sepsis following methylprednisolone: restoration of beta-adrenergic receptor responsiveness? PMID- 9742539 TI - Salicylate toxicity masquerading as malignant hyperthermia. AB - We report a case of hyperpyrexia presumed due to topical salicylate toxicity occurring immediately following general anaesthesia for appendicectomy in an eleven year old boy. Some of the features strongly suggested the diagnosis of malignant hyperpyrexia. PMID- 9742540 TI - Intraoperative QRS-interval changes caused by hyperkalaemia in an infant with Arima syndrome. AB - A one-year-and-ten-months-old male infant with Arima syndrome, a very rare genetic disorder, underwent urgent insertion of a catheter for continuous ambulatory peritoneal dialysis (CAPD) under general anaesthesia. During the procedure he showed QRS-interval changes caused by hyperkalaemia which was successfully treated with calcium gluconate. The management and intraoperative complications of this syndrome are reported and available literature reviewed. PMID- 9742541 TI - Difficult airway in a patient with Marshall-Smith syndrome. AB - Marshall-Smith syndrome is a rare clinical disorder characterized by accelerated bone maturation, dysmorphic facial features, airway abnormalities and death in early infancy because of respiratory complications. Although patients with Marshall-Smith syndrome have several features with potential anaesthetic problems, previous reports about anaesthetic management of these patients do not exist. We present a case, in which severe hypoxia developed rapidly after routine anaesthesia induction in an eight-month-old male infant with this syndrome. After several unsuccessful attempts the airway was finally secured by blind oral intubation. After 2 weeks, laryngeal anatomy was examined with fibreoptic laryngoscopy which revealed significant laryngomalacia. Laryngoscopy was performed without problems with ketamine anaesthesia and spontaneous breathing. The possibility of a compromised airway should always be borne in mind when anaesthetizing patients with Marshall-Smith syndrome. Anaesthesia maintaining spontaneous breathing is safest for children with this syndrome. If tracheal intubation or muscle relaxation is required, precautions are needed to maintain a patent airway. Muscle relaxants should possibly be avoided before intubation. PMID- 9742542 TI - Prolonged mivacurium neuromuscular block in children. AB - The authors report two cases of prolonged neuromuscular block after administration of mivacurium in children with previously undiagnosed plasma cholinesterase deficiency related to homozygous atypical genotype. Their anaesthetic management is described as well as determination of the phenotype of both children and their family. PMID- 9742544 TI - Anaesthesia in a child with deletion 13q syndrome. PMID- 9742543 TI - Use of remifentanil in infants. AB - We describe the use of remifentanil in three infants with complex medical issues (hepatic failure, cyanotic heart disease and renal compromise). The short duration of opioid effect even after a long period of drug infusion (18 h) suggests this drug may be useful in some infants. Continued study is warranted. PMID- 9742545 TI - Edward's syndrome (trisomy 18) PMID- 9742546 TI - Paracetamol pharmacokinetics in the premature neonate; the problem with limited data. PMID- 9742547 TI - Difficult airway management in a baby with Axenfeld-Rieger syndrome. PMID- 9742548 TI - Challenge of pharmacodynamic variability for drug delivery and pharmaceutical technology. PMID- 9742549 TI - Transdermal permeation modulation by cyclodextrins: a mechanistic study. AB - The purpose of this study was to investigate permeation modulation by beta- and 2 hydroxypropyl-beta-cyclodextrins (beta-CD and HP-beta-CD, respectively) alone and complexed with penetration enhancers for the test drugs 5-fluorouracil and estradiol through human skin, and to probe the value of the CDs in a barrier cream against toluene exposure. Methods include phase solubility studies, permeation experiments, and thermal analysis of stratum corneum; inclusion complexes were characterized by Karl Fischer titrimetry, infrared spectroscopy, and thermal analysis. Results show that complexes of terpenes or toluene with beta-CD were insoluble, whereas those with HP-beta-CD were soluble. The CDs did not enhance flux of either the polar or lipophilic drugs through skin; estradiol permeation was reduced following membrane pretreatment with either CD. Complexation of the lipophilic terpenes with the CDs reduced enhancer efficacy. When formulated into a barrier ointment both CDs, but particularly beta-CD, retarded toluene permeation through the skin and delayed the onset of maximum flux. It is concluded that the CDs themselves are not penetration enhancers for 5 fluorouracil or estradiol in human skin, and that they may be usefully incorporated into a barrier formulation to reduce percutaneous absorption of toxic materials on occupational exposure. PMID- 9742550 TI - Production of microparticles by high-pressure homogenization. AB - A high-pressure homogenization method for the production of aqueous suspensions of poly(D,L-lactide) and poly(D,L-lactide-co-glycolide) was investigated. Depending on the production conditions it was possible to produce micro--as well as nanoparticulate systems without the use of organic solvents. The influence of different homogenization temperatures and different dispersion media on particle size and charge was investigated. Additionally, various polymer/surfactant ratios were investigated. Homogenization in phosphate buffer at temperatures above the glass transition temperature (Tg) of the polymers resulted in wide particle size distributions with a high percentage of particles in the nanometer range. The effect of a second homogenization step at a temperature below Tg was examined at two different homogenization pressures. Additional homogenization cycles at 10 degrees C led to smaller particle size distributions and the average particle sizes were smaller. The stability of the particles was affected by the concentration of surfactant as well as by the zeta potential of the particles. Phosphate buffer kept the pH of the suspension in a range that provided a high surface charge of the particles because of deprotonization of the carboxylic functions of the polymer. PMID- 9742551 TI - Binding, molecular mechanics, and thermodynamics of cyclodextrin inclusion complexes with ketoprofen in aqueous medium. AB - The purpose of this work was to study the interaction forces involved in the inclusion processes of ketoprofen with several cyclodextrins and to assess the best cyclodextrin for complexing this anti-inflammatory drug. The behavior of the inclusion complexes of ketoprofen with alpha-, beta-, and gamma-cyclodextrins was studied by UV-VIS direct spectroscopy, 1H NMR, and molecular mechanics. Thermodynamic parameters for the binding processes were obtained from the temperature variations in binding constants, which manifest that "nonclassical" hydrophobic interactions are the main forces involved in these inclusion processes. Binding constants show that beta- and gamma-cyclodextrins form more stable 1:1 complexes with ketoprofen than does alpha-cyclodextrin. 1H NMR spectra show that the inclusion degree depends on the size of the internal diameter of cyclodextrin. The geometries calculated on the bases of molecular mechanics for these three-dimensional models indicate high stability. PMID- 9742552 TI - Purified guar galactomannan as an improved pharmaceutical excipient. AB - The purpose of this study was to assess certain pharmaceutical attributes of guar galactomannan, a hydrocolloid polysaccharide obtained from the endosperm of the leguminous plant Cyamopsis tetragonolobus (L.), following purification using both literature procedures and new processes. Experiments were performed to measure viscosity, hydration rate, tablet hardness, and dissolution profiles of guar galactomannan both before and after purification. The viscosity of an aqueous 1% purified galactomannan solution is typically 40-50% higher than its unpurified guar galactomannan precursor. The hydration rate of an aqueous 1% purified galactomannan solution increases by 100% after purification. These physicochemical changes resulted in improvements in pharmaceutical properties such as better stir speed independence in both tablet and capsule dissolution profiles and improved tablet hardness. For instance, time to 50% dissolution of ranitidine HCl from capsules containing unpurified guar gum was 0.4 and 1.8 hr at 20 and 40 rpm, respectively, using USP Apparatus II. Using the same amount of purified guar gum and the same conditions (20 and 40 rpm), these values were increased to 2.9 and 3.8 hr, respectively. These data demonstrate a reduced effect of changing agitation conditions and the need for less guar gum to sustain the release of a water-soluble drug. Tablet hardness of purified guar gum (particle size < 75 microns) was about 7 kP and the same unpurified guar gum of equal particle size and hydration gave a hardness of less than 1 kP. PMID- 9742553 TI - Formulations of sugars with amino acids or mannitol--influence of concentration ratio on the properties of the freeze-concentrate and the lyophilizate. AB - Formulations consisting of either sucrose or trehalose with glycine, lysine-HCl, or mannitol were studied to determine how the ratio of the excipients affects the design of the lyophilization program and the properties of the final cake. Glass transitions (Tg', Tg), crystallization temperatures, and eutectic melting temperatures were measured by differential scanning calorimetry, the physical state of the excipients was determined by x-ray powder diffraction, and residual moisture was measured by Karl Fischer titration. The addition of increasing amounts of glycine, lysine-HCl, or mannitol to a sucrose solution caused a progressive depression of the Tg', an effect that was more pronounced with the amino acids. In equivalent ratios with sucrose, the two amino acids induced a comparable Tg' shift due to their low Tg' values. For lysine-HCl, two apparent Tg' with midpoint temperatures of -69 and -56 degrees C were measured. For mannitol, the Tg' depression was unexpected because mannitol exhibits a higher Tg' than that of sucrose. During lyophilization, the ratio of the amorphous amino acids or mannitol to the sugar determined whether crystallization could be induced by an annealing step performed after freezing. Crystallization could be verified by a shift of the formerly depressed Tg' back to the value of the sugar and by the detection of the eutectic melting peak of the crystallized compound. The crystallized excipients served as excellent bulking agents. In the freeze dried cake, amorphous glycine and even more amorphous mannitol lowered the Tg value. If the cake was stored above Tg, subsequent crystallization of mannitol occurred. The results emphasize that the qualitative and quantitative composition of a formulation has profound implications on the design of a lyophilization program and on the characteristics of the freeze-dried cake. PMID- 9742554 TI - Effects of formulation and process variables on the aggregation of freeze-dried interleukin-6 (IL-6) after lyophilization and on storage. AB - This study assessed the impact of residual moisture, Tg, and excipient physical state of different formulations on the "in-process" and shelf-life stability of freeze-dried interleukin-6 (IL-6). The effect of an annealing procedure was also evaluated. Characterization of the lyophilizates was done by Karl Fischer titration, differential scanning calorimetry (DSC), and x-ray measurements. Analysis of protein stability was carried out by size exclusion chromatography (SEC), sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and turbidity measurements. During freeze-drying, the most effective protection against aggregation was provided by completely amorphous formulations consisting of trehalose or sucrose either alone or in combination with glycine or mannitol. Other amorphous formulations like those of sucrose with lysine-HCl or dextran could not provide comparable stabilization. In lyophilizates containing a crystallized excipient such as glycine or mannitol, IL-6 suffered destabilization, which was less pronounced if an additional amorphous excipient was present. For the completely amorphous formulations, aggregation was prevented during a 9-month storage at 25 and 40 degrees C as long as the storage temperature did not exceed the Tg value of the lyophilizate, otherwise severe damage occurred. Formulations containing amorphous dextran or lysine-HCl could not effectively stabilize IL-6 even when stored below Tg. Annealing helped to improve cake robustness and appearance, but for lyophilizates containing an excipient crystallized by annealing an increase of IL-6 aggregation was observed despite a storage below Tg. Thus, the amorphous state of the excipients and a high Tg can be considered necessary conditions for preventing aggregation of freeze-dried IL-6. Whether the conditions are also sufficient depends on the choice of excipients. Destabilization can occur with some excipients despite their amorphous state as well as in the presence of crystallized excipients despite a storage below Tg. Compared to sucrose, trehalose is a more favorable excipient for protein lyophilization because it exhibits a higher Tg, possesses better stabilizing properties, and can reduce protein aggregation which may have been caused by annealing. PMID- 9742555 TI - Polysaccharide film-coating process for freely swellable hydrogels. AB - In order to control the drug release from coated hydrogels by preventing membrane fractures, an intramembrane freely swellable matrix device was designed by enclosing a void space between a crosslinked poly(vinyl alcohol) (PVA) matrix and a calcium alginate membrane. The highly swellable PVA matrix loaded with diltiazem hydrochloride was obtained by means of a simplified procedure of the polymer crosslinking reaction using glutaraldehyde in solution with ammonium persulfate. The undried swollen matrix was coated with a calcium alginate membrane employing an ionotropic gelation of sodium alginate induced by calcium ions. The subsequent drying process generated a void space separating the inner core from the membrane. The resulting calcium alginate membrane, which was uniform and compact in the structure, increased in thickness according to the coating time. Coating times exceeding 5 min allowed modification of the drug release profile providing, after a short burst period, sustained and constant rate phases in both simulated gastric fluid and simulated intestinal fluid. Because the inner hydrogel expanded freely inside the device, the unstressed and intact membrane could act as the rate-controlling factor in the drug release process. Owing to the pH-dependent behavior of the membrane, most of the drug was delivered in intestinal fluid. Therefore, the device proposed could be advantageously used for drug targeting to the small intestine. PMID- 9742556 TI - Preparation and evaluation of beads made of different calcium alginate compositions for oral sustained release of tiaramide. AB - This study was undertaken to develop a sustained-release formulation of tiaramide (TAM), a non-steroidal anti-inflammatory drug with a short half-life, using alginate of different chemical compositions. Alginate gel beads containing TAM were prepared using a gelation of alginate with calcium cations. Bead performance was evaluated in vitro for different dissolution media and beads were also subjected to coating. TAM release was dependent both on its solubility in dissolution medium and the guluronate residue content of the alginate used. The release rate was in the following order: in pH 1.2 > pH 6.8 > water. The fast release rate in pH 1.2 is the result of the high solubility of TAM in acidic medium. Beads based on alginate rich in guluronate residue had the lowest release rate, which can be attributed to the compact structure formed by guluronate residues through cooperative interaction with calcium ions. Alginate beads were administered to beagle dogs, and pharmacokinetic parameters (mean residence time [MRT], tmax, Cmax, and AUC) were calculated. In vivo results were in good agreement with in vitro dissolution characteristics. Beads with high guluronate content gave the best controlled results. In addition, coated beads showed a more satisfactory sustained-release pattern. Calcium alginate appears to be a potential carrier for controlling drug release rate, even for water-soluble drugs such as TAM. PMID- 9742557 TI - Development of a highly efficient purification process for recombinant adenoviral vectors for oral gene delivery. AB - Recently, replication-deficient adenoviruses have received increasing attention as vector for gene delivery and as potential vaccine carriers. With the increased use of the vector in vivo and in clinical trails, the demand for a safe, rapid, and cost effective purification process has been heightened. In this report, a simple and efficient method for the purification of large quantities of live adenoviral vectors was developed. The process involved the replacement of cesium chloride (CsCl) gradients with sucrose gradients. Ultracentrifugation times were reduced and the desalting step eliminated, decreasing total preparation time by 15 hr. A 20-80% linear sucrose gradient provided optimal recovery of infectious viral particles and positioning of the viral band in the gradient. Purification with this gradient system produced a preparation containing 1.39 x 10(14) lac forming units (lfu)/ml. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis revealed that the process also removed all associated cellular proteins from the preparation. Studies have shown that direct lyophilization of the vector in sucrose after purification produces a product containing 1.4 x 10(12) lfu/ml. Minimal degradation was seen in the lyophilized preparation. A viral concentration of 6 x 10(11) lfu/ml was detected in the product after 150 days in storage at -20 degrees C. This approach will not only simplify the preparation of adenoviral vectors for in vivo studies and clinical trials, but will facilitate production of stable adenoviral formulations for oral gene delivery. PMID- 9742558 TI - Factors that influence stability of recombinant adenoviral preparations for human gene therapy. AB - This report identifies formulation and processing factors that influence stability of viral preparations such as selection of appropriate buffer systems, cryoprotectants, and cooling rates. Adenovirus type 5 containing the lacZ marker gene was suspended in combinations of trehalose, sorbitol, sucrose, mannitol, glycine, CaCl2, and gelatin. X-gal stains of 293 cells were used to determine the lac-forming units (lfu)/ml of each preparation before and after treatments. Phosphate-buffered solutions (except those containing sucrose or trehalose) demonstrated a drop of 3 pH units upon freezing regardless of cryoprotectant used. Tris-buffered solutions demonstrated a variation in pH which was dependent upon chosen cryoprotectant, with 1 M trehalose exhibiting no change and a 5% mannitol/10 mM CaCl2 combination showing a 3-unit drop in pH. 4-[2-Hydroxyethyl] 1-piperazine ethanesulfonic acid (HEPES)-buffered solutions showed little change in initial pH when frozen regardless of cryoprotectant chosen. In solution, adenovirus was not affected by incubation for 24 hr in buffers ranging from pH 4 to 8. However, when the solutions were frozen, the number of remaining infectious virions was dependent upon the final pH of the suspending medium. Cryoprotectant solutions that significantly maintained viral stability during a single freeze- thaw cycle were 0.5 M sucrose, 0.5 M trehalose, and 10% sorbitol/0.4% gelatin. Long-term stability studies were performed at 4 degrees C with lyophilized sorbital/gelatin and sucrose preparations. Both formulations provided adequate stability for the adenovirus, with 2.6 and 5.6 x 10(11) lfu/ml detected 150 days after drying, respectively. PMID- 9742559 TI - Intranasal vaccination: pharmaceutical evaluation of the vaccine delivery system and immunokinetic characteristics of the immune responses. AB - The purpose of this study was to analyze the effect of some pharmaceutical excipients when used for mucosal vaccine formulations and to characterize the achieved immune response. After conducting various pharmaceutical evaluations of the formulations, immunokinetic studies were performed in mice, guinea pigs, and rabbits. The kinetics and the characteristics (antibody isotypes, etc.) of the immune response were studied, as well as the induced level of toxin neutralizing IgG antibodies, which are usually used as the only measures of the potency of vaccines. Results in mice show that intranasal vaccination results in a potent and rapid immune response, similar to that seen after subcutaneous immunization. In guinea pigs and rabbits, however, the subcutaneous immunization produced significantly stronger response than did intranasal vaccination. The most promising excipients were found to be either Polysorbate 20 or Cremophor EL in an aqueous mixture together with caprylic/capric glyceride. The results indicate that nontoxic and pharmaceutically acceptable excipients can be used for mucosal vaccination, providing an interesting alternative to parenteral vaccination. PMID- 9742560 TI - Complexation and solubility behavior of albendazole with some cyclodextrins. AB - The main purpose of this work was to study the albendazole-cyclodextrins complexation equilibrium and to propose a suitable excipient to improve the solubility behavior and dissolution rate of albendazole. The complexation of albendazole with four cyclodextrins, alpha-CD, beta-CD, gamma-CD, and hydroxypropylated (HP)-beta-CD, has been studied by using electronic absorption spectroscopy and molecular mechanics. The equilibrium was studied at pH 7.5 under various temperature conditions, and at pH 1.8 with HP-beta-CD at 298 K. The albendazole binding constant was the greatest for the HP-beta-CD. Both the un ionized (Alb) and the ionized species (AlbH+) were shown to interact with HP-beta CD. The studies at different temperatures suggest that the hydrophobic effect is the most important driving force in these systems. Moreover, the dissolution rate studies with beta- and HP-beta-CDs in the buffered aqueous solution at pH 7.5 have been accomplished and the dissolution rate was observed to increase with the cyclodextrin concentration. The solubility behavior was studied with the Higuchi and Connors method. The phase solubility and direct spectroscopy methods reveal a 1:1 inclusion complex in all of the studied cases. Molecular mechanics data show the most probable structure of the complexes. PMID- 9742561 TI - Comparative dissolution studies for mefenamic acid-polyethylene glycol solid dispersion systems and tablets. AB - The purpose of this study was to enhance the dissolution of mefenamic acid (MFA) through the formation of solid dispersion systems, and to compare the dissolution of the unformulated dispersions with those of formulated dispersions in tablets. Solid dispersions of MFA were prepared in polyethylene glycol 3350 (PEG) as a binary system, and PEG and Tween 20 (TW) as a ternary system by the melt method. The dispersions were characterized by dissolution, scanning electron microscopy, and powder x-ray diffraction studies. A decrease in MFA composition in the binary dispersion systems from 50 to 5% w/w resulted in a 50% increase in the dissolution rate during the period of study, and this was threefold higher than that of pure MFA. Incorporation of TW in the preparation of ternary dispersion systems resulted in a further increase in MFA dissolution. A sevenfold increase in MFA dissolution was observed when the ternary system composition was MFA/PEG/TW 4.7:93:2.3 (% w/w). Scanning electron microscopy and x-ray diffraction pictures showed an increase in size and decrease in crystallinity of the dispersions, respectively. Compression of the dispersions into tablets did not have any effect on the dissolution of the drug from the dispersions. Compression of pure MFA and Avicel PH 101, which was used as a diluent and disintegrant, resulted in a threefold increase in dissolution. However, the dissolution of the uncompressed mixture was identical to that of pure MFA. Thus, further processing of the solid dispersions into tablets did not decrease the rate of dissolution of the drug in the dispersions. This may be very important in the formulation of solid dispersions as tablets, which could lead to a reduction in the dose of practically water-insoluble drugs. PMID- 9742562 TI - A methodology for the optimization of wet granulation in a model planetary mixer. AB - This paper investigates a methodology for the optimization of wet granulation processes in planetary mixers. A model formulation was granulated in a planetary mixer (two different bowl sizes). The wet masses were characterized by their bulk density and consistency (as measured by mixer torque rheometry), and the feasibility of scale-up from one mixer bowl to the other was studied using a dimensionless numbers approach for the estimation of the power consumption at the granulation end point. Both bowls gave the same dimensionless power relationships (a relationship between the power number, Reynolds number, Froude number, and bowl fill ratio), which could therefore be used for calculating the power consumption level when the wet mass achieves its target values of density and consistency, i.e., the point at which granulation should be stopped. It was also shown that batches granulated in different conditions (batch size, blade speed) in two planetary mixers, but presenting similar wet mass characteristics (bulk density and consistency) led to dry granules of similar properties: granule size distribution, density, friability, and flow. This work suggests that it is possible to characterize the wet mass by only two parameters which describe the quality of the downstream granules. The scale-up procedure based on the use of dimensionless numbers was found to be applicable to planetary mixers, provided they give one common dimensionless power relationship. PMID- 9742563 TI - Fetal outcome and maternal morbidity after early amniocentesis. AB - We have studied a large population-based cohort of women who had amniocentesis at 14 weeks' gestation (early amniocentesis) in Victoria, a state of Australia, to determine fetal loss rates and maternal morbidity. This was done by linking two registers--one containing information on all prenatal diagnostic tests in the state, and the other a register of all births at or after 20 weeks' gestation. Almost complete follow-up was achieved. The spontaneous fetal loss rate was significantly higher for women of age 37 years and over having early amniocentesis (2.5 per cent), as compared with the fetal loss rate found previously on the same geographically defined population who had amniocentesis at about 16 weeks' gestational age (1.1 per cent). Three classes of maternal morbidity reported to the birth register (post 20 weeks' gestation) were also analysed. The most significant finding was a reduced rate of premature rupture of membranes with early amniocentesis when compared with a group having later amniocentesis, or the background population not having any amniocentesis. There was no significant increase in the occurrence of antepartum bleeding or genito urinary tract infection for women having early amniocentesis. These data agree with other studies in showing that early amniocentesis is associated with a significant increased risk of fetal loss, as compared with later amniocentesis. In addition we have shown no significant increase in the occurrence of three indicators of maternal morbidity, reported at or after 20 weeks' gestation. PMID- 9742564 TI - Early amniocentesis: effect of removing a reduced volume of amniotic fluid on pregnancy outcome. AB - In mid-trimester amniocentesis (MTA), 12-15 ml of amniotic fluid is aspirated for cytogenetic analysis. When a similar volume of amniotic fluid is removed by early amniocentesis (EA), it represents a significant proportion of the total amniotic fluid volume in the first trimester. The fluid depletion, which may persist for 7 to 10 days, is considered to impair development of fetal lungs and extremities and, possibly, contribute towards procedure-related congenital abnormalities and miscarriages. By only removing 7 ml of amniotic fluid, we have demonstrated a total miscarriage rate (3.8 per cent) comparable with previous large studies (Table V), a low incidence of respiratory difficulties at birth (2.7 per cent) and a low incidence of fixed flexion deformities (1.6 per cent), at the expense of a small increase in the incidence of culture failure (2.2 per cent). PMID- 9742565 TI - Prenatal diagnosis of isolated hypospadias. AB - Ten cases of prenatal diagnosis of isolated hypospadias are presented, six of them in the second trimester. The echographical basis for the suspected diagnosis of hypospadias are: anomalous distal morphology of the penis, small lateral folds (dermal remains of the prepuce), small penis with ventral incurving and anomalous urinary stream. The embryogenesis and the clinical utility of prenatal study of the genitals of the fetus, not only to determine the sex, but also to detect anomalies, are discussed. PMID- 9742567 TI - Evaluation of prenatal diagnosis of congenital heart disease. AB - Prenatal diagnosis performed by fetal ultrasound scan is now a routine part of antenatal care in many countries. That an increasing number of fetal anomalies may be detected on prenatal ultrasound is beyond doubt. What is possible is not, however, always practical, especially where congenital heart diseases (CHDs) are concerned and when whole antenatal populations are screened rather than high risk groups. Thanks to our registries of congenital anomalies, a retrospective study was undertaken to evaluate the prenatal detection of CHDs by ultrasound scan in 92,021 consecutive pregnancies of known outcome from 1990 to 1993. Only 107 out of 779 malformed fetuses with CHDs without chromosomal anomalies were detected (13.7 per cent). The sensitivity of detection varied from 50 per cent for malformations, such as hypoplastic left heart and single ventricle, to around 5 per cent for ventricular and atrial septal defects. The effectiveness of the detection of some forms of major congenital heart disease has increased since 1987 by including routine examination of the four-chamber view and of the inflow and outflow tracts of the fetal heart. Our results stress the need to obtain a definite, clear, four-chamber view, to perform scans at > 18 weeks' gestation and to train sonographers in order to improve prenatal detection of CHDs. PMID- 9742566 TI - Steroid sulphatase deficiency is the major cause of extremely low oestriol production at mid-pregnancy: a urinary steroid assay for the discrimination of steroid sulphatase deficiency from other causes. AB - A method for determining whether a pregnant woman with an extremely low serum oestriol (ELSE) measurement of mid-trimester is carrying a fetus with steroid sulphatase deficiency or another more serious disorder is described. We undertook GC/MS analysis of steroids in random maternal urine samples and quantified oestriol, oestriol precursors (dehydroepiandrosterone (DHEA), 5-androstene-3 beta, 17 beta-diol, 16 alpha-hydroxy-dehydroepiandrosterone and 5-androstene-3 beta, 16 alpha, 17 beta-triol), pregnanediol, and five other steroids largely unaffected by pregnancy (androsterone, etiocholanolone, tetrahydrocortisol, 5 alpha-tetrahydrocortisol and tetrahydrocortisone). Thirty-two samples collected from seven normal pregnant women between the 7th and 27th week of pregnancy and 22 from individuals with ELSE were analysed. Diagnostic ratios of excreted products were developed. These included ratios of oestriol and oestriol precursors to the cumulative value for the five non-pregnancy-related steroids and ratios of oestriol and oestriol precursors to pregnanediol and to each other. Our data demonstrated high 3 beta-hydroxy-5-ene steroid excretion in all ELSE patients together with low urinary oestriol excretion, a situation only consistent with deficiency of steroid sulphatase. The normal individuals had high oestriol and low excretion of oestriol precursors. No patient in our series showed the low oestriol levels and low oestriol precursor values that would indicate a fetal adrenal abnormality as the underlying defect. PMID- 9742569 TI - Coping with serum screening for Down syndrome when the results is given as a numeric value. AB - Forty-six pregnant women undergoing second-trimester biochemical screening for Down syndrome were asked to fill in the State-Trait Anxiety Inventory (STAI) questionnaire to assess their anxiety level at two different moments: when recruited to the study (at 11-13 weeks' gestation), and after the test result was communicated. The test result was given as a numeric value of risk (1/x), rather than as positive/negative. There were 10 women in whom the risk after biochemical screening increased (median delta risk = +1/535, range = 1/69 to 1/1083), whereas in the remainder the risk decreased (median delta risk = -1/1576; range = -1/142 to -1/4947) compared with the baseline value calculated on maternal age alone. Although only in a minority of women the STAI score after biochemical screening exceeded the reference range, the change in the STAI score was significantly higher when the risk increased, and the change in the risk estimate correlated significantly with the change in this index of anxiety. Three out of seven women with a 'negative' test, but increased risk estimate and increased anxiety after biochemical screening chose to undergo amniocentesis. A policy of providing the result of biochemical screening for Down syndrome as a numeric value, even for 'negative' tests, may cause some women to experience anxiety and request amniocentesis. PMID- 9742568 TI - Detection of fetal congenital heart disease in a low-risk population. AB - Our purpose was to evaluate the efficacy of level two ultrasound screening for the detection of congenital heart defects (CHD) in a low-risk population by using three standardized cuts. Within a period of four years a total of 6727 pregnant women of a low-risk population undertook several ultrasound examinations on the basis of screening for fetal malformations. All ultrasound examinations were performed by three experienced doctors. At every single scan three standardized cuts (apical and lateral four-chamber view, crossing over of the great arteries) were obtained in order to detect congenital heart defects. Of 87 CHDs (1.33 per cent of the examined women) 39 (43.8 per cent) were diagnosed prenatally. The detection rate was 10/48 (20.8 per cent) in the presence of VSD, ASD2 or combined ASD2 + VSD, the detection rate was 29/39 (74.3 per cent) in the presence of other forms of congenital heart disease. None of the 38 missed cases in the first group but three of the ten missed CHDs in the second group had emergency neonatological problems. Aneuploidy and/or other malformations existed in 22/87 cases of CHD. The obstetrical management was changed in nearly all cases after the diagnosis of a CHD. Twenty-two women opted for termination of pregnancy because of additional fetal malformations or chromosomal defects. Five women were transferred prenatally to a tertiary centre for neonatal cardiac surgery. Ten deliveries were performed in the presence of a neonatologist. Good detection rates for CHD can be achieved in a low-risk population on the basis of level two ultrasound screening by using the above mentioned three cuts and thus, the perinatal mortality and morbidity can be improved. PMID- 9742570 TI - Beta-glucuronidase P408S, P415L allele in a Mexican population: population screening in Guadalajara and prenatal diagnosis. AB - Recently we identified a P408S, P415L allele of beta-glucuronidase in several Mexican patients with mucopolysaccharidosis type VII (Sly syndrome) and presented evidence that both mutations are required to produce the MPS VII allele (Islam et al., 1996). In an attempt to determine whether either of these mutations exists as a benign polymorphism among Mexicans, we developed a PCR method to screen simultaneously for both mutations and used it to screen a population sample of 187 Mexican individuals in the Guadalajara area, all from the north-western states of Mexico. Neither mutation was present in 374 alleles studied. In addition, we had the opportunity to carry out prenatal diagnosis in a fetus at risk for homozygosity for this MPS VII allele at the 15th week of gestation using enzymatic assays as well as by analysis of genomic DNA isolated from cultured amniotic fluid cells. The fetus was found to be heterozygous for the P408S, P415L allele. The newborn's heterozygosity was confirmed after birth by enzyme assays on plasma and leukocytes, and by analysis of DNA from leukocytes. PMID- 9742571 TI - Prenatal diagnosis for keratin mutations to exclude transmission of epidermolytic hyperkeratosis. AB - Epidermolytic hyperkeratosis (bullous congenital ichthyosiform erythroderma) is an autosomal dominant skin disorder caused by mutations in keratins 1 and 10. We have used direct gene sequencing to ascertain the status of a 15 week fetus of parents whose first child was affected with this disorder. The parents show no clinical signs of epidermolytic hyperkeratosis but were concerned about the possibility of transmitting the disorder due to germline mosaicism. Molecular analysis of the affected son revealed a G to A mutation in codon 156 of keratin 10, resulting in an arginine to histidine substitution within the highly conserved 1A region. Codon 156 has been previously identified as a mutational hot spot and substitutions of this arginine residue are very common in epidermolytic hyperkeratosis patients. Analysis of genomic DNA isolated from amniotic cells showed that the fetus did not harbour this mutation and a healthy infant was eventually born that was unaffected by this disorder. PMID- 9742572 TI - Prenatal diagnosis of Friedreich ataxia. PMID- 9742573 TI - Prenatal diagnosis of genetic syndromes may be facilitated by serendipitous findings at fetal blood sampling. AB - Two women without a specific risk had fetuses with multiple malformations diagnosed by ultrasound; extensive biochemical investigations on fetal blood revealed clues which would have allowed the correct diagnosis of a genetic condition: Pallister-Killian syndrome in one with increased fetal LDH, and Smith Lemli-Opitz type II syndrome in the other with low fetal cholesterolaemia. When compared with chorionic villus sampling and amniocentesis, rapid karyotyping in women with multiple fetal malformations by fetal blood sampling allows the collection of additional data which may lead to the diagnosis of specific genetic syndromes. PMID- 9742574 TI - In utero exchange transfusion in homozygous alpha-thalassaemia: a case report. AB - We report a case of homozygous alpha-thalassaemia with hydrops fetalis presenting at 22 weeks of gestation. In utero exchange transfusion was performed with maternal blood at 23 weeks. 25 weeks and 29 weeks of gestation. The fetus was delivered at 29 weeks of gestation without significant neonatal complication. Post-transfusion haemoglobin pattern after transfusion suggested that a total haematocrit of 0.52 may be the desired post-exchange transfusion haematocrit to aim for and the total haematocrit of haemoglobin A and haemoglobin Portland dropped approximately one percent per day. PMID- 9742575 TI - Alternative methods of maternal weight adjustment in maternal serum screening for Down syndrome and neural tube defects. AB - Serum markers used in screening for Down syndrome and neural tube defects are often adjusted to take account of the effect of maternal weight on the marker levels. The standard adjustment procedure is based on a linear relationship between the marker concentration, expressed as the log of the multiple of the median (MOM), and maternal weight on a linear scale. It has been proposed that maternal weight adjustment may be better performed using a linear relationship between marker concentration expressed in MOM and the reciprocal of maternal weight. In a dataset of 8905 singleton pregnancies in white women without Down syndrome or neural tube defects we compared the two methods of weight adjustment and found that both were satisfactory and neither had an obvious advantage over the other. In the analysis it was noticed that hCG levels in very heavy women (> 120 kg) were higher than expected from the decreasing linear trend with maternal weight--a result that was statistically highly significant (p < 0.01) but for which we have no explanation. In screening it will have virtually no effect because the finding was restricted to only the 0.3 per cent of the heaviest women. PMID- 9742576 TI - The pattern of maternal serum inhibin-A concentrations in the second trimester of pregnancy. AB - Maternal serum inhibin-A concentration is a useful marker in prenatal screening for Down syndrome in the second trimester. We measured inhibin-A concentrations in 4304 pregnancies without Down syndrome between 14 and 22 weeks of pregnancy to determine the median values according to gestational age. There was a U-shaped pattern of inhibin-A concentration against gestational age with a minimum concentration at 17 weeks and 1 day (120 days). We suggest that screening centres use a quadratic equation when estimating their normal median inhibin-A level, constraining the shape of the curve and fixing the lower point of the curve to occur at 120 days, but allowing its position in relation to the vertical inhibin A axis to be set according to the local data. This approach will systematically allow for the changing inhibin-A concentration without introducing the instability of deriving a fresh quadratic equation for each screening centre and each time a centre's medians are revised. PMID- 9742577 TI - Region-specific FISH probes used to identify and characterize an interstitial paracentric inv(21)(q22.1q22.3). AB - Region-specific probes developed for the diagnosis of specific syndrome, can be adapted to elucidate the exact nature of certain chromosomal structural anomalies. We describe the use of FISH probes in characterizing a prenatally diagnosed chromosome rearrangement. An abnormal chromosome 21 was detected during amniocentesis for maternal age indication, and a similar appearing chromosome 21 was found in the mother. The exact nature of the rearrangement was not immediately evident from G-banded karyotypes. FISH was performed using a whole chromosome painting probe, as well as the region-specific probes D21S65 (21q21 22.1), D21S55 (21q22.3) and D21S1219/D21S1220 (21q22.3-qter) (Oncor). Results showed an interstitial paracentric inversion, with breakpoints in bands 21q22.1 and 21q22.3, which was identical in the mother and the fetus: 46,XX,?inv(21)(q).ish inv(21)(q22.1q22.3)(wcp+.D21S65 mv, D21S55 mv, D21S1219/D21S1220 st). In this case, FISH using region-specific probes was helpful in characterizing the inversion and aided in the genetic counselling of risk assessment for the family. PMID- 9742578 TI - Ultrasound prenatal diagnosis of the Nail-Patella syndrome. AB - The Nail-Patella syndrome (NPS) is an autosomal dominant connective-tissue disorder characterized by the absence or hypoplasia of the nails and patella, posterior illiac horns, elbow deformities, congenital nephropathy cervical ribs and eye problems. The presence of the posterior iliac horns is pathognomonic and have been observed in more than 80 per cent of cases. In this report, we present the first case of prenatal diagnosis of NPS by ultrasound. The possible kidney involvement, combined with other, milder, complications make the prenatal diagnosis of this syndrome worthwhile. PMID- 9742579 TI - Prenatal diagnosis and outcome of mosaicism for a de novo unbalanced translocation identified in amniocytes. AB - Mosaicism for an unbalanced reciprocal translocation was identified in cultured amniocytes of a 16-week-old fetus; mos46,XX,der(4)t(4;5)(q34;q12)/46,XX. Parental karyotypes were normal, indicating a de novo origin of the unbalanced translocation in the fetus. The additional chromosomal material on the der(4) was derived from chromosome 5 as demonstrated by both GTG banding and fluorescence in situ hybridization with a chromosome 5 paint. Two subsequent amniocenteses, at 18 and 20 weeks, confirmed the presence of the abnormal cell line. A percutaneous umbilical blood sample (PUBS) contained only normal cells, 46,XX, and a high resolution ultrasound revealed no fetal abnormalities or growth retardation. The pregnancy was continued and a normal female was born at term. No evidence of the unbalanced translocation cell line was found in cord blood or placental samples at birth. The finding of mosaicism for an unbalanced translocation at amniocentesis is rare, and is associated with a high risk of fetal abnormality. This case illustrates the importance of follow-up studies by PUBS and high resolution ultrasound for further assessing the risk of phenotypic abnormality. PMID- 9742580 TI - Fluconazole teratogenicity. PMID- 9742581 TI - Polyhydramnios as a first prenatal symptom of non-ketotic hyperglycinaemia. PMID- 9742582 TI - Tetraploidy in a growth-retarded fetus with a thick placenta. PMID- 9742584 TI - Anencephaly in monozygotic twins and recurrence risk. PMID- 9742583 TI - False-negative results of trisomy 21 on direct analysis on chorionic villus sampling. PMID- 9742585 TI - Reaching the fetal environment: a tribute to DR Hermogenes Alvarez. PMID- 9742586 TI - Current awareness in prenatal diagnosis. PMID- 9742587 TI - [Dependence of learning characteristics on visual object properties in Rhesus macaca by bilateral removal of the 7th field of the parietal cortex]. AB - Removal of the rhesus monkey parietal cortex 7th field exerted no effect on learning processes involving visual discrimination of images united in their colour and geometrical form, but the learning of differentiating the spatial information did suffer. The data obtained suggests that, in the process of learning visual differentiation, spatial differentiating signs are formed, the process involving neuronal structures of the 7th field of the inferior cortex. Removal of the 7th field disrupts mechanisms of the body scheme assession and egocentric orientation resulting from visual-vestibular interrelationships. PMID- 9742588 TI - [Modulation of the long-term memory by delayed administration of the nootropic agent L-pyroglutamyl-D-alaninamide during spaced and massed training of rats]. AB - L-pyroglutamyl-D-alanine amide significantly enhanced freezing response in the group of rats with massed training and reduced it in the group with spaced training. The control animals revealed a higher freezing response with the spaced training. No differences occurred between the groups in a cue test. PMID- 9742589 TI - [Neuron mechanisms of the regulation of bulbar vagosolitary activity by structures of basolateral amygdala nuclei in cats]. AB - Neuronal reactivity to single stimuli applied to both the peripheral nerves and the cortex, was the same in the cat solitary tract nucleus. The 1-20 Hz stimulation frequency rendered the reactivity either tonic in character or with reduced firing rate. A high degree of convergence (80.2%) on baso-lateral nuclei neurons was established, as well as a significant cortico-fugal effect (71%) on the viscero-sensory units of these nuclei. Single shocks and tetanic stimulation of the baso-lateral amygdala evoked mostly phasic responses of the primary as well as secondary vagal neurons. Feed-back mechanisms of the amygdaloid control of the bulbar viscero-sensory units are discussed. PMID- 9742590 TI - [Effect of the intermittent hypoxic training on the functioning of peptidergic neurons of the paraventricular hypothalamic nucleus and brain stem neurons in rats]. AB - Internittent hypoxic training (IHT) increased the quantity and secretory activity of peptidergic neurons of the paraventricular hypothalamic nucleus (PHN) and activated neurons of the dorsal motor nucleus of n.vagus. These structures seem to take part in realisation of the IHT activating effect on condition of the pancreatic delta-cells. The effect involves insulin-stimulating and insuloprotective effects realised via hypothalamic and neuro-conducting ways of regulation of the endocrine pancreas with a direct participation of hypothalamic neuropeptides. PMID- 9742591 TI - [The activity of neurons of the thalamus and brain cortex hemispheres during EEG complexes "slow wave-spindle wave" in rabbits]. AB - Neurons of the medial thalamus and some cortical neurons revealed the same activity upon the spindle wave as upon the slow wave. The membrane potential was changing in a similar way. The data obtained suggests that the slow wave mechanism is analogous to the mechanism of the spindle waves, whereas the rhythm of complexes occurrence or their common with the "spontaneous" sleep spindles rhythm, are determined by a different mechanism. PMID- 9742592 TI - [Participation of rostral ventral medullary neuron structures in the regulation of the respiratory rhythm mechanism in rats]. AB - The role of neuronal structures of the rostral ventromedullary area in regulation of central inspiratory activity, was studied. The data obtained suggests that the structures of subretrofacial area are important for the respiratory rhythm generation due to regulation of excitability of the inspiratory neurons. PMID- 9742593 TI - [Recovery of the gas exchange and oxygen transport after long-term hypothermia in rats]. AB - After a deep prolonged hypothermia, fast warming up of experimental rats (0.26 degree C/min) to the body temperature 35-36 degrees C entailed recovery of the heart rate, blood pressure, 1-min blood volume, oxygen consumption in rats. However, in the process of warming up, cardiac output, function of the left ventricle, oxygen consumption and the CO2 production were decreased. Acidosis and haemoconcentration following a 3 hr prolonged hypothermia were reversible. PMID- 9742594 TI - [Recovery of the oxygen consumption, oxygen transport, and cardiac activity after hypothermia followed by respiration and cardiac arrest in rats]. AB - After a 1 hr deep prolonged immersion hypothermia, warming up of experimental rats in water (the temperature increase 0.25 degree C/min) entailed a complete recovery of all the parameters of gas exchange, haemodynamics, external respiration, and the blood. After a 3 hr hypothermia, under the same conditions of warming up, oxygen consumption, CO2 production, and 1 min blood volume were obviously decreased, and the animals died within 0.5-1.0 hr. PMID- 9742595 TI - [Effects of diazepam and piracetam on the rat behavior responses after combined exposure to the low doses of ionizing radiation and heat]. AB - Effects of low doses of ionizing radiation and heat on behavioural responses of white rats and effects of diazepam and pyracetam long after separate and combined exposure to the two factors were studied. These changes were not present in a month after cessation of the exposure. Certain features of the modulating effect of pyracetam on behavioural responses, were revealed. PMID- 9742596 TI - [The role of thyroid hormones in prevention of disorders of myocardial contractile function and antioxidant activity during heat stress]. AB - The stress of heat under conditions of immobilisation induced an obvious depression of the cardiodynamic parameters. This correlated well with intensification of lipoperoxydation and a drop in the myocardial antioxydant activity. Small doses of thyroid hormones prevented the decline of the parameters, normalisied myocardial free-radical homeostasis in result of activation of superoxyddysmutase, catalase, and general antioxydant activity. PMID- 9742597 TI - [Changes in the blastogenic lymphocyte transformation during kindling induced by picrotoxin in rats]. AB - Picrotoxin-induced kindling was shown to suppress the blastogenic response to bacterial lipopolysaccharide and phytogemagglutinin in male Wistar rats. The delta-sleep-inducing peptide as well as carbamazepine prevented the epileptogenic effects of picrotoxin. Carbamazepine was also effective against decreasing of phytogemagglutinin-induced blastogenic response. PMID- 9742598 TI - [The role of mesotocin in the regulation of osmotic permeability of the urinary bladder epithelium in the frog]. AB - The addition of 0.01 nM to 0.1 nM mesotocin to serosal Ringer solution of the frog urinary bladder did not change the osmotic water permeability, but increased hydroosmotic effect of 0.01 nM arginine vasotocin or 10 nM desmopressin. The data obtained allow one to conclude that mesotocin is involved in the modulation of the osmotic water permebility in frog urinary bladder. PMID- 9742599 TI - [Mechanisms of the reproductive system aging and hormonal carcinogenesis: modification caused tobacco smoke exposure]. PMID- 9742600 TI - [Changes in the number of corticosterone receptors in the rat brain after neonatal hydrocortisone administration induced by unavoidable stress]. PMID- 9742602 TI - [Mechanisms of the microcirculation during hypoxia induced by reduced local pressure]. PMID- 9742601 TI - [Determination of the temperature of thermosensitive spots of the human skin]. PMID- 9742603 TI - [Analysis of the electroencephalogram based on a modified amplitude-interval algorithm]. PMID- 9742604 TI - [Digital image processing in physiological studies]. PMID- 9742605 TI - [Pre-Botzinger complex participates in respiratory effects of thyroliberin]. AB - Responses of the TRH microinjections into the pre-Botzinger complex of adult anesthetised rats involved a dose-dependent increase in respiratory rate as well as shortening of inspiratory and expiratory duration. The tidal volume and inspiratory muscles' activity decreased following high concentrations of the TRH. The heart rate did not change. The findings suggest that the pre-Botzinger complex is at least partly responsible for the TRH tachypnoic effect. PMID- 9742606 TI - [Participation of reticular neurons of the cat medulla oblongata in the integration of the respiration center activity]. AB - Reticular neurons of the respiratory centre are divided into three groups. The 1st one is located in the medial areas and accepts afferents from chemoreceptors. The 2nd one is located in inspiratory and expiratory areas of the centre and take part in integration of afferent signals and activation of the effector mechanism. The 3rd group is present in both areas and organizes the effector activity of the centre. Electrical stimulation of the brain stem nuclei can modify reticular neurons in lateral areas. PMID- 9742607 TI - [Comparative analysis of changes in respiration and systemic hemodynamics during activation of GABA receptors in cats and rats]. AB - Effects of alpha-hydroxybutyrate (GHB) involved a decrease in the respiration rate and subsequent periodic breathing in anesthetised cats and rats. No considerable changes occurred in systemic circulation. There were, however, some differences between the species in the patterns of respiratory arrest. Possible mechanisms of the similarity and difference are discussed. PMID- 9742608 TI - [The role of pharyngeal muscles in the regulation of airway resistance during inspiration resistive load in rabbits]. AB - In anesthetised spontaneously breathing rabbits, an increase in the inspiratoryactivity m.genioglossus was greater than a simultaneous increase in the respiratory activity of the diaphragm cluring loaded breathing. A bypass of the air flow via tracheostoma did not affect the response of the m.genioglossus to a resistive load. The difference seems to be due to different functional inputs from the lung mechanoreceptors to phrenic and hypoglossal neuronal pools. PMID- 9742609 TI - [Effects of the local stimulation of the insular cortex on respiration in rats]. AB - Two distinct patterns of respiratory responses to electrical microstimulation were elicited from the rat insular cortex: a decrease in respiratory airflow and tidal volume with no alteration of the respiratory rate (the inhibition response) and an increase in respiratory rate and inspiratory airflow (the excitation response). Gastric motility changes could also be elicited from the anterior insular cortex simultaneously with the inhibition response. The data suggests overlapping of the inhibition response area in the anterior insular cortex with a gastrointestinal representation there, and of the excitation response area in the posterior insular cortex with a cardiovascular representation. PMID- 9742610 TI - [Comparative analysis of responses of the thoracic and abdominal respiration components to hypercapnia and muscular work]. AB - The ratio of thoracic and abdominal contribution to ventilatory responses to hypercapnia in humans was found to depend on the initial ratio of thoracic and abdominal breathing reserves as determined by the body position in space. In muscular work, participation of the respiratory muscles in locomotor loads becomes a factor affecting the ratio of thoracic and abdominal components of ventilatory response. PMID- 9742611 TI - [Mediators of the local reflex arc in airways of the rat and guinea pig]. AB - Transmission of neural pulses from afferents to functional module's neurons in rats and guinea pigs can be carried out with the aid of the P substance. Further transmission to a smooth muscle of the trachea occurs through a neuro-muscular junction with acetylcholine as a transmitter. Transmission of inhibitory influences from the tracheo-bronchial receptors to the smooth muscle is carried out via nicotinic cholinoreceptors. Adrenergic mechanisms can modulate neural transmission at the level of intramural ganglia. PMID- 9742613 TI - [Characteristics of the response area of mouse inferior colliculus neurons within the critical band]. AB - The inferior colliculus neurons which characteristic frequencies are in the range of 17-25 kHz, corresponding in mice to the width of psychophysical critical band with central frequency of 20 kHz, revealed all types of rate/level functions traditional to the auditory system neurons: monotonic, plateau and non-monotonic. Simultaneous presentation of two tones provide the following effects in the neuron response area: threshold sensitivity shift to high intensities; the reduction of intensity range within the neuron frequency response area; the increase of neurons selectivity to the stimulus frequency parameters, cause not only restriction of response exhibitory area, but threshold decrease on the CF stimulus. The finding suggest a coexistence of two inhibitory systems, one of which provides the stability of auditory system's frequency selectivity in the wide intensity range and the other regulating perception of signal intensity within neuron frequency area. PMID- 9742612 TI - [Pain sensitivity during chronic psychoemotional stress in humans]. AB - In a natural "model" of a chronic psychoemotional stress in humans, (neurasthenia) a decrease in the nociceptive threshold was found. The decrease resulting from weakening of supraspinal tonic descending inhibition which, in its turn, evidently was due to a diminished activity (or dysfunction) of the brain opioid system occurring in a long-lasting psychoemotional stress. PMID- 9742614 TI - [Development of the vascular system in brain hemispheres in developing rats]. AB - The blood passage from a major artery to a major vein was digitally reconstructed as well as hydraulic pressure, linear blood flow velocity, wall shear stress, and vascular resistance, following postmortem studies of 2-week old, 1-month old, and adult Wistar rats. The findings suggest that cerebro-cortical vascular bed acquires its functional specifics by the 3rd-4th week after birth. PMID- 9742615 TI - [Modulation of the antiarrhythmic effect by endogenous opioids during adaptation to hypoxia in rats]. AB - Adaptation of rats to hypoxia increased the heart resistance against arrhythmogenic effects. The Met-enkephaline-Arg6-Phe7 contant increased in the hypothalamus of the adapted animals. The blockers of mu- and delta-opioid receptors reduced the antiarrhythmic effect of the adaptation, whereas chi receptor inhibitor, nor-binaltorphimine, completely abolished it. The findings suggest the antiarrhythmic effect depends on an increase of the endogenous opioids level and modulated effect of these peptides upon autonomous nervous system. PMID- 9742616 TI - [Effect of the unilateral ovariectomy in rats on the brain development in one month old offspring]. AB - Litter of unilaterally ovariectomised rats had a greater mass of the brain and a thicker cortex of the anterior parietal lobe as well as a lower density of neurons distribution in this lobe. The brain changes seem to be due to changes in the hormones level in operated rats. PMID- 9742617 TI - [Changes in the number of CD4+ T-helpers in the bone marrow of aggressive mice of C57BL/6J and CBA strains]. AB - Aggressive behaviour led to immunostimulation with a subsequent rise in the number of T-helpers in C57B1 and CBA mice. The number of CD4+ T-helpers in their bone marrow correlated with duration of successive experience of victories. The effect of such a behaviour on immunity seems to be due to an activation of the dopaminergic system. PMID- 9742618 TI - [Effect of opioid peptides on immunomodulation]. AB - Activation of the opioid receptors by delta-agonist DSLET and by kappa-agonist rimorphin led to a significant inhibition of plaque-forming and rosette-forming cells in the CBA mice. On the other hand, mu-agonist DAGO stimulated the immune response on the 4th and 5th days after immunization with SRBC (5 x 10(8)). Lesion of the hypothalamo-hypophyseal connection prevented immuno-stimulating as well as immuno-depressive effects. The latter seems to be due to an interaction with the serotoninergic system, whereas immuno-stimulating effect involves the dopaminergic system. PMID- 9742619 TI - [Analysis of the body heat condition during muscular work based on the heat physical model of the rabbit body]. AB - Quantitative data on body weight, heat absorption and heat conduction, value of relative body surface, and other values reflecting the main heat-physical parameters of the rabbit body, were obtained for a heat-physical model. The model "body" revealed a comparatively great heat inertia capacity which explains a passive rise of the "body" temperature in light or moderate physical work. Conditions of increasing the heat-physical efficacy of thermoregulatory vascular responses in animals, were revealed. PMID- 9742620 TI - [Effect of enkephalins on the liver secretory function]. AB - A decrease in the bile flow and bile acids, proteins and lipids secretion in enkephalines infusion into the portal vein was revealed. The bile secretion changes under the effect of enkephaline seem to result from the latter effect upon metabolic processes in hepatocytes. PMID- 9742621 TI - [Bile secretion induced by L-thyroxine and substance P in dogs]. AB - I.M. administration of L-thyroxin activated hepatic secretory function and changed the bile chemical contents. The substance P intensified production of bile and increased the absolute contents of free and conjugated bile acids, cholesterine, bilirubin, and total protein in the bile. PMID- 9742622 TI - Co-culture update: creating an embryotrophic environment in vitro. AB - As infertility treatments evolve and techniques are developed to improve the fertilization and pregnancy rates, it is clear that the in vitro environment in which the gametes/embryos are cultured is less than perfect when compared to the in vivo counterpart. This becomes a serious problem for couples who seek to have children but are limited by unsuccessful attempts to become pregnant and a technology that is behind in mimicking the in vivo environment that would enhance gamete interaction and embryo development. In an attempt to begin to correlate the in vivo and in vitro microenvironment of the gametes/embryos and to recognize the potential of the fallopian tube as more then a highway for oocytes to transverse in their pathway to the uterus, coculturing of tubal cells along with gametes and embryos was introduced to the in vitro fertilization community. This report is an attempt to compile the research and results of those attempting to improve pregnancy rates by improving in vitro culture conditions via co-culture of cells with gametes and/or embryos. PMID- 9742623 TI - Embryo cryopreservation. AB - Cryopreservation stands as an ongoing evolution in the field of assisted reproductive technologies. Face with increasing numbers of fertilized oocytes and early embryos, cryopreservation avails the ART program of a useful means to preserve embryos for future use without exposing patients to the risks of multiple pregnancies. This article examines some of the clinical and laboratory issues critical to a successful cryopreservation program. PMID- 9742624 TI - Optimization of culture conditions for human in vitro fertilization and embryo transfer. AB - Approximately 20 years ago the first child conceived with in vitro fertilization (IVF) and embryo transfer was born in England. Although overall pregnancy rates and delivery rates after IVF have improved over the years, success between IVF clinics can vary as much as tenfold. The factors that influence the success rate include type of patient, ovarian stimulation protocol, quality of oocytes, culture conditions, handling of gametes and embryos, quality of embryos, embryo transfer technique, and endometrial receptivity. Culture conditions and handling of gametes and embryos will be reviewed with the hope of improving success of human IVF. The ongoing clinical pregnancy rates in our program increased from about 30% in 1995 and 1996 to about 50% in 1997. Improved embryo culture conditions and embryo cryopreservation technology should not only increase fresh but also frozen-thawed embryo transfer pregnancy. This will make IVF efficient and cost-effective while achieving high overall pregnancy rates with lower multiple pregnancy rates. PMID- 9742625 TI - An update on human fertilization. AB - The process of fertilization and the role that each gamete plays in that process have been the subject of investigation in a large number of species and for many years. However, while much is now known for some species relatively little is known for others. Indeed, the specific events that are required to occur in the human male and female gametes and that facilitate fertilization are still somewhat ill-defined. For example, as of today, there have been numerous biomolecular processes that have been put forth as playing an important role in the sequence of events during which sperm acquire fertilizing ability, yet the actual significance of many of these remains suspect. This article will summarize what is presently best known about prefertilization processes occurring in human spermatozoa. For those interested in nonhuman mammalian and nonmammalian species, articles addressing this and other topics can be found in the many review articles cited herein. PMID- 9742626 TI - Integrins, endometrial maturation, & human embryo implantation. AB - Cell-cell and cell-extracellular matrix interactions are fundamental processes involved in cell migration and tissue remodeling. Both the cyclic regeneration of the human endometrium during the menstrual cycle as well as the process of embryo implantation involve such dynamic interactions. It has become quite clear that integrin adhesion molecules expressed on the surface of cells play critical roles in the transmission of signals from the extracellular milieu to the cells. It is these signals that presumably regulate the behavior of these cells during major morphogenetic processes. In recent years, work in human endometrium and trophoblasts has uncovered both the regulated and constitutive expression of integrin subunits and their extracellular matrix ligands in these tissues. In addition, attempts have been made to correlate pathological states related to either infertility or abnormal pregnancy to the aberrant expression of several of these integrins. The purpose of the present review is to describe briefly our present state of knowledge of the expression of integrins in human endometrium and trophoblasts and provide the reader with the necessary background needed to understand, at the cellular and molecular levels, processes in reproduction such as embryo implantation. PMID- 9742627 TI - [The biological characteristics and treatment of acute promyelocytic leukemia]. PMID- 9742628 TI - [The clinical significance of MDR-1 gene expression in the hemopoietic cells of patients with acute leukemias in different phases of the disease]. AB - AIM: Analysis of cytostatic therapy effects on expression of gene MDR-1 in hemopoietic cells of patients with acute leukemia (AL) in complete clinicohematological remission (CCHR). MATERIALS AND METHODS: The study included 48 AL patients. 27 of them were untreated, 25 were resistant to or had recurrent AL. 4 patients were followed up. Bone marrow mononuclear fraction was investigated. Expression of MDR-1 gene in the cells was evaluated at hybridization. RESULTS: High expression of MDR-1 gene occurred in leukemic blast cells either upon achievement of CCHR or at least 6 months after its onset. When using schemes of chemotherapy containing two potential inductors of gene MDR-1, expression of this gene was registered significantly more frequently than in using schemes based on one inducing drug (p < 0.05). Frequency of occurrence of enhanced expression of gene MDR-1 in leukemic blasts significantly correlated with frequency of CCHR (p < 0.05). Correlation between occurrence of the gene's expression in normal hemopoietic cells in CCHR and occurrence of early AL recurrences was not found. CONCLUSION: The findings may facilitate design of new AL treatment programs. PMID- 9742629 TI - [The empirical antibiotic therapy of patients with acute leukemias: the results of a multicenter study]. AB - AIM: To evaluate efficacy of ampicilline/sulbactame and fluconasole in the regimen of empirical antibiotic therapy in patients with acute leukemia. MATERIALS AND METHODS: The trial covered 14 hematological departments of Russia and 1 of Ukraine. Acute myeloid leukemia patients were included. 92 cases of fever in 56 patients with analysis of efficacy in 66 cases were considered. At the first stage of empirical antibiotic therapy, cefoperason (4 g/day) and gentamycin (240 mg/day) were administered. If no response was reached, ampicilline/sulbactam (7.5 g/day) was added. This was the second stage. If no response occurred for 5 days the three drugs were joined by fluconasol (400 mg followed by 200 mg). RESULTS: Fever of unclear genesis was cured in 82% (28 of 34), clinical infection--in 80% (20 of 25), microbiologically confirmed infection -in 4 of 7 cases. A complete response to the empirical antibiotic therapy was registered in 52 of 66 cases (79%). 7(10.5%) patients died of infectious complications. 7(10.5%) received other antibiotics. PMID- 9742630 TI - [An evaluation of the prognostic significance of antigen CD95(Fas/APO-1) expression on the cells of patients with a myelodysplastic syndrome, acute myeloid leukemia and chronic myeloleukemia]. AB - AIM: The expression of CD95(Fas/APO-1) antigen was studied on bone marrow cells of 19 MDS patients, peripheral blood blast cells of 15 acute myeloid leukemia (AML) patients, blast cells and granulocytes of 68 patients with chronic myeloid leukemia (CML)--24 in chronic, 9 in accelerated phase and 35 in blastic crisis (BC)--by indirect surface immunofluorescence assay using flow cytometry (FACScan, Becton Dickinson, USA). RESULTS: CD95(Fas/APO-1) antigen was revealed on bone marrow cells of 8 out of 19 (36.8%) MDS patients; the percentage of antigen positive cells was 38.1 +/- 19.2%; on 45.5 +/- 22.8% of cells in 6(45%) of 15 AML patients. Fas/APO-1 antigen was totally absent in CML chronic stage; its expression was found in 34% (12 of 35) of our patients with CML BC on peripheral blood blasts and in 56% (5 of 9) on peripheral blast cells of CML patients in acceleration phase. CONCLUSION: The data on overall survival of CD95-positive MDS patients suggest that the presence of Fas antigen is a favorable prognostic sign for patients with MDS. The patients from CD95-negative group represent a risk group both for survival and AML transformation. In CML BC group the survival does not depend upon Fas-antigen expression. PMID- 9742631 TI - [Hyperexpression of the multiple drug resistance gene (MDR-1) in chronic myeloleukemia patients]. AB - AIM: To elucidate prognostic value of MDR-1 gene expression in patients with chronic myeloid leukemia (CML). MATERIALS AND METHODS: The MDR-1 gene expression was studied by in situ hybridization in hemopoietic cells of 63 Ph-positive CML patients in different phases of the disease. The survival of the patients and duration of the chronic phase (CP) were evaluated using the Caplan-Meyer method. RESULTS: MDR-1-positive patients had a shorter survival (p < 0.01) and CP (p < 0.05) than negative ones. MDR-1 gene overexpression has no impact either on the survival or duration of AP and BP (p < 0.05). Moreover, the MDR-1 gene overexpression is not dependent either on the previous treatment or other prognostic markers. CONCLUSION: Overexpression of MDR-1 gene is an independent prognostic factor and an additional parameter to Sokal's scores. PMID- 9742633 TI - [Lymphocyte morphology]. PMID- 9742632 TI - [The hypereosinophilic variant of Ph'-positive chronic myeloleukemia]. AB - AIM: To confirm clonal nature of idiopathic hypereosinophilic syndrome (IHES), its relevance to Ph'-positive chronic myeloid leukemia. MATERIALS AND METHODS: 3 cases of idiopathic hypereosinophilic syndrome are reported with morphologic analysis of bone marrow cells and cytogenetic examinations. In one patient the presence of Ph'-chromosome was confirmed at fluorescent in situ hybridization (FISH) and molecular-genetic analysis (bcr/abl). Samples of bone marrow, spleen and liver were examined pathohistologically. RESULTS: The presence of chromosome anomaly t(9;22), i.e. Ph'-chromosome, associated with chronic myeloid leukemia (CML) was identified in all the 3 cases. There was also myeloid hyperplasia in the bone marrow (with primarily mature, eosinophilic granulocytes), spleen and liver, depression of megakaryocyto- and erythropoiesis. 2 patients had similar clinical symptoms which was not typical for CML in chronic phase: fever, elevated ESR, clear-cut anemia and thrombocytopenia. In the absence of hyperleukocytosis, blood and bone marrow eosinophils remained high (42.5, 21.5, 42.5% and 21.4, 7.1, 6.5%, respectively) due to "mature" forms. The number of blasts in the bone marrow was maximum 2.4%. CONCLUSION: The literature and the obtained data suggest closeness of idiopathic hypereosinophilic syndrome and Ph'-positive CML within myeloproliferative diseases. PMID- 9742634 TI - [Treatment results in children with B-cell lymphomas using a modified BFM protocol]. AB - AIM: Trial of the modified protocol BFM in the treatment of children with B-cell lymphomas. MATERIALS AND METHODS: 26 children with B-cell lymphoma were treated. Of them 2 children were treated according to the program for risk 2 group and 24 children were treated as risk 3 group. RESULTS: Complete remission was achieved in 23 patients. Two children were resistant. There were 2 cases of early recurrence, 1 case of early death, 1 case of death in remission. 20 patients are in complete remission. The 4-year survival is 78%. CONCLUSION: The modified protocol BFM proved to be highly effective against B-cell lymphoma in children. Its drawback is high toxicity. PMID- 9742635 TI - [The role of HLA antigens in predicting the active course of multiple myeloma]. AB - AIM: Detection of associations between carrying some HLA-antigens class I in patients with multiple myeloma (MM) and activity of the malignant process. MATERIALS AND METHODS: 76 MM patients received polychemotherapy. Its efficacy was assessed after one, three, six and twelve courses by reduced blood and/or urine levels of monoclonal protein, signs of bone healing, reestablishment of normal number of plasma cells in the bone marrow. Identification of HLA-antigens was made in two-stage lymphocytotoxic complement-mediated test using the standard panel of the anti-HLA sera. Data on HLA-typing of 865 blood donors served control. The findings were statistically processed. RESULTS: All the patients were divided into 3 groups: with indolent (n = 18), active (n = 25) and aggressive (n = 37) MM course. In patients with aggressive MM course high chi square values were estimated for three HLA specificities: HLA-B13, HLA-B40, HLA B5. Only HLA-B13 proved significant. No significant differences in carrying HLA antigens were revealed for patients with active MM course. CONCLUSION: The survival of MM patients depends on the degree of the malignant process activity. Patients with aggressive MM course significantly more frequently carry HLA-B13, therefore it can be considered a genetic marker of MM. Its detection can serve a criterion for determination of adequate polychemotherapy. PMID- 9742636 TI - [The efficacy of polychemotherapy programs in treating multiple myeloma patients]. AB - AIM: To compare efficiency of the programs MCVP, VCAP and ARA-COP in the treatment of multiple myeloma (MM) as regards completeness of the response, duration of the remission and toxicity. MATERIALS AND METHODS: A total of 41 MM patients entered the study (27 females, 14 males, age from 41 to 72 years, MM duration from 1 month to 8 years). 16, 10 and 15 patients were treated according to MCVP, VCAP and ARA-COP programs. RESULTS: Both in the resistant and primary patients the response was the highest to ARA-COP treatment. The remission or stabilization was achieved in 93.4% of patients. VCAP program was less effective. However, clinicohematological remission was achieved in 50% of patients. This program is rather heart toxic. MCVP program was the least effective. Survival was followed up in 16 patients (10 MCVP, 3 VCAP and 3 ARA-COP patients). The survival was 20-62, 16-36, 23.6-64.8 months for ARA-COP, VCAP and MCVP, respectively. CONCLUSION: ARA-COP program proved most effective of the three programs both in primary and drug-resistant patients. VCAP and MCVP programs are less effective but can be used in primary management of MM patients. PMID- 9742637 TI - [The late results of the combined therapy of patients in 2nd-stage lymphogranulomatosis]. AB - AIM: To define the scope of combined therapy according to prognostic factors in patients with Hodgkin's disease state II. MATERIALS AND METHODS: 98 patients with favorable and unfavorable Hodgkin's disease (HD) prognosis according to EORTC criteria (41 and 57 of group 1 and 2, respectively) entered the study. Unfavorable factors were: mixed HD variant and lymphoid depletion, ESR above 50 mm/h in stage A and 30 mm/h in stage B, involvement of more than 3 groups of lymph nodes, age over 40. Patients of group 1 received CVPP program: 2 courses before and after radiation of the primary disease zones in the total dose 40 Gy. Therapy of group 2 patients consisted of 3 CVPP courses before and 3 courses after irradiation of all the lymphatic collectors above the diaphragm in the total dose 35 Gy or radiation according to the extended program. Efficacy of therapy was assessed by EORTC criteria. The survival curves were calculated by Caplan and Meyer methods. RESULTS: In groups 1 and 2 a complete a complete remission was achieved in 98 and 93%, 6-year survival was 100 and 91%, recurrence free survival--94 and 87%, respectively. Survival free of the treatment failure reached in group 1--88%, in group 2--81%. CONCLUSION: Reduced treatment in HD stage II in favourable prognosis did not worsen the results of treatment. PMID- 9742639 TI - [Total irradiation of the liver in treating patients with lymphogranulomatosis]. AB - AIM: The study of efficiency of radiation in specific affection of the liver in lymphogranulomatosis (LGM). MATERIALS AND METHODS: 212 LGM patients were examined for hepatic lesions using x-ray, radiological, cytochemical and histological investigations of the biopsies. Coagulation, alkaline phosphatase, copper and ceruloplasmin in the serum were measured to evaluation the disease activity. All the patients received polychemotherapy (PCT). Irradiation of the liver in the total focal dose 36-44 Gy was performed in confirmed involvement of the liver as adjuvant to PCT. RESULTS: Specific liver lesions were detected in 6.1% of patients at primary examination and in 15.6%--at follow-up. Liver lesions occurred most frequently in mixed-cell variant of LGM and lymphoid depletion. Eradication of liver lesion after PCT was achieved in 15.4%. Subsequent radiation produced a complete remission in 100% of patients with diffuse and 80% of patients with large-focal liver lesion. In detection of liver involvement at the time of LGM progression eradication and remission were achieved in 70 and 50% of patients, respectively. From 30% of patients with liver lesions resistant to PCT and radiation, 15% had stable disease. The survival did not depend on eradication of liver lesion. CONCLUSION: In involvement of the liver its radiation is indicated for all the patients with new-onset LGM. In the recurrence, liver radiation is recommended for those in whom the liver is the only extranodal lesion. PMID- 9742638 TI - [The factors of hereditary predisposition to lymphogranulomatosis]. AB - AIM: To determine genetic factors of predisposition marked HLA with reference to serological and molecular characteristics. MATERIALS AND METHODS: Four groups of patients were included in the study: 51 patients with lymphogranulomatosis (LGM), 33 healthy relatives of these patients, 37 patients with chronic myeloid leukemia (CML), 24 healthy relatives of these CML patients. 224 donors served control. HLA antigens were identified with the lymphocytoxic test and PSR-MSSR. Results of typing of class II antigens were taken into consideration in coincidence of serological and DNA typing. The significance of the differences was estimated according to the chi-square criterion. RESULTS: Differences in frequency of distribution of HLA-antigens (subloci A and B) were not found. Antigen CW7 was present in 70, DR5 in 60, DR6 in 50% of LGM patients. This frequency was much higher than in control groups. Carriers of CW7 are at 7 times higher risk to develop LGM. Among LGM patients the number of homozygous individuals is higher than in healthy ones (50 and 15.6%) while the number of individuals with a complete set of HLA-A.B antigens is significantly less. CONCLUSION: Genetic predisposition to LGM is predetermined by HLA antigens CW7, DR5, DR6. Genes HLA DR1 and HLA-DR7 protect carriers from factors provoking LGM. Common HLA genes in the parents predispose their children to LGM. Insufficiency of the phenotype is a factor predisposing to LGM. PMID- 9742640 TI - [A trial of using allogeneic bone marrow transplantation in children with different hematologic neoplastic diseases]. AB - AIM: To define optimal time for transplantation of bone marrow (TBM) in children with hematological malignancies. MATERIALS AND METHODS: 20 allogenic TBMs were performed in children with acute myeloblastic leukemia (6 patients, 2 of them in recurrence), acute lymphoblastic leukemia (7 patients, 4 of them in recurrence), chronic myeloid leukemia (CML) in a chronic stage (3 patients), severe aplastic anemia (3 patients), generalized neuroblastoma (1 patient). Pretransplantation preparation included cyclophosphamide and busulphane or cyclophosphamide, busulphane and vepezide. The graft-versus-host reaction (GVHR) was prevented with cyclosporin A plus methotrexate or cyclosporin A plus urbazone. Engrafting was recognized by change of karyotype and blood group. RESULTS: From 13 children with acute leukemia subjected to TBM in a complete remission 4(33%) are alive, 5 died within 100 days after TBM (TBM was made in recurrence in 4 children), 3 patients died of recurrence 12 months after TBM. One patient with CML and one with severe aplastic anemia remain in remission. The main complications and causes of death in early posttransplantation period were hemorrhagic syndrome, infectious complications, GVHR. According to a one-year follow-up, the recurrent disease caused death most frequently. CONCLUSION: Positive result of TBM is related to the disease stage at transplantation. PMID- 9742641 TI - [The transplantation of hemopoietic cells in patients with solid tumors]. AB - AIM: Evaluation of efficiency of bone marrow and peripheral stem cells transplantation in patients with solid tumors. MATERIALS AND METHODS: A total of 38 patients aged 30-40 years with stage III or IV solid tumors (15 cases of breast cancer, 5 cases of Ewing's sarcoma, 4 cases of osteogenic sarcoma, ovarian or testicular tumors, 3 cases of soft tissue sarcoma) underwent high-dose polychemotherapy with autologous hematopoietic rescue. All the patients were resistant to the first line chemotherapy or were in relapse of the disease. 21, 11 and 6 patients were transplanted bone marrow, peripheral blood stem cells and both bone marrow and peripheral blood stem cells, respectively. RESULTS: The only factor which influenced overall survival of the patients was the stage of the disease at the moment of the transplantation. 66% of patients in complete or partial remission survived 3 years versus 16.6% of those in progression or relapse of the disease. Other parameters such as the patients' age, the source of hemopoietic cells, number of chemotherapy courses were not essential for the survival. 2 patients died after transplantation. The main cause of death was relapse or progression of the disease after the transplantation. CONCLUSION: Transplantation of hemopoietic cells is a promising therapy of patients with solid tumors sensitive to chemotherapy. PMID- 9742642 TI - [The transfusion aspects of allogeneic bone marrow transplantation]. AB - AIM: Analysis of transplantations of allogenic bone marrow (BM) for determination of transfusiological problems arising in various types of donor's and recipient's blood ABO incompatibility. MATERIALS AND METHODS: 46 allogenic BM transplantations from relative donors are analysed. The following types of ABO incompatibility were identified: significant (12 donor-recipient pairs), insignificant (10 pairs), combined (2 pairs). RESULTS: The proposed methods of BM fractionation provided the yield of nucleated cells in minimal admixture of red cells. This allowed to minimize loss of stem cells at fractionation and to prevent acute hemolysis at infusion of BM suspension in cases of ABO group different BM allotransplantations. CONCLUSION: The proposed modification of the "shelf" method enabled effective fractionation of small BM volumes obtained from children-BM donors with high yield of nucleated cells and small admixture of incompatible red cells. Reinfusion of erythrocytic mass obtained at fractionation of donor BM fractionation used as replacement therapy makes allogenic blood transfusion unnecessary in 66% of BM donors. PMID- 9742643 TI - [Comparative data on the use of a cryosupernatant and fresh-frozen plasma in the therapy of a protracted infectious-septic syndrome of disseminated intravascular coagulation]. AB - AIM: To compare therapeutic effectiveness of cryosupernatant plasma fraction (CSP) and fresh-frozen plasma (FFP) in long-term infectious-septic DIC syndrome arising in acute abscess and gangrene of the lung. MATERIALS AND METHODS: 106 and 131 patients with infectious-septic DIC syndrome were treated with CSP and FFP, respectively. The results of the treatment were compared clinically and hemotopogically. RESULTS: Clinical response to both treatments was evident from positive changes in XIIa-dependent fibrinolysis, lowering of fibrinogen level and enhanced activity of antithrombin III. CSP treatment brought about a decrease in the number of thromboses, lethal cases, unfavorable outcomes. CONCLUSION: The cryosupernatant can be used instead of fresh-frozen plasma in combined treatment of long-standing infectious-septic DIC syndrome. PMID- 9742644 TI - [The blood serum albumin transport function in patients with acute and chronic leukemias]. AB - AIM: The study of changes in functional activity of albumin ligand centers in the course of accumulation of metabolic products in the blood of hematological patients. MATERIALS AND METHODS: Blood serum albumin transport function (TF) was investigated in 22 and 24 patients with acute and chronic leukemia, respectively. Lipid analyser AKL-01 and the kit "Zond-Albumin" ("Zond", Moscow) were employed to assess total albumin concentration (TAC), effective albumin concentration (EAC), albumin binding reserve (ABR), the toxicity index (TI). The control group consisted of 38 blood donors. The findings were processed with statistical techniques. RESULTS: In patients with manifest acute leukemia, ABR and TI decreased while TAC remained unchanged. Chronic leukemia patients had high EAC and close to control levels of ABR and TI. In terminal chronic leukemia ABR lowered, TI increased, TAC was high but EAC fell. In patients with chronic leukemia and acute leukemia remission changes in albumin TF was accompanied with growing TAC. CONCLUSION: A decline of albumin TF was found in patients with manifest acute leukemia and terminal chronic leukemia. In non-terminal chronic leukemia and in remission of acute leukemia albumin TF was normal. PMID- 9742645 TI - [An unusual effect of cyclosporine A in a patient with chronic lympholeukemia complicated by autoimmune hemolysis and thrombocytolysis]. PMID- 9742646 TI - [The adult respiratory distress syndrome during the recovery of the neutrophil level after autologous bone marrow transplantation]. PMID- 9742647 TI - [Extramedullary lesions in acute myeloblastic leukemia]. PMID- 9742649 TI - [Current antihistamine preparations]. PMID- 9742648 TI - [The treatment of infections in patients with neutropenia (a review of the literature with the inclusion of the authors' own data)]. PMID- 9742650 TI - Effect of oral probenecid coadministration on the chronic toxicity and pharmacokinetics of intravenous cidofovir in cynomolgus monkeys. AB - In animals and humans, intravenous administration of the antiviral nucleotide analogue cidofovir results in a dose-limiting nephrotoxicity characterized by damage to the proximal tubular epithelial cells. Probenecid, a competitive inhibitor of organic anion transport in the proximal tubular epithelial cells, was evaluated for its effect on the chronic toxicity and pharmacokinetics of cidofovir. Cynomolgus monkeys (5/sex/group) received cidofovir for 52 consecutive weeks as a once weekly intravenous bolus injection at 0 (saline), 0.1, 0.5, or 2.5 mg/kg/dose alone or at 2.5 mg/kg/dose in combination with probenecid (30 mg/kg/dose via oral gavage 1 h prior to cidofovir administration). Cidofovir associated histopathological changes were seen only in the kidneys, testes, and epididymides. Nephrotoxicity (mild to moderate cortical tubular epithelial cell karyomegaly, tubular dilation, basement membrane thickening) was present only in monkeys receiving 2.5 mg/kg/dose cidofovir without probenecid. The incidence and severity of testicular (hypo- and aspermatogenesis) and epididymal (severe oligo- and aspermia) changes were increased in monkeys administered cidofovir at 2.5 mg/kg/dose, either alone or in combination with oral probenecid. Renal drug clearance was decreased between Weeks 1 and 52 in the 2.5 mg/kg/dose groups and resulted in an increased systemic exposure to cidofovir (as measured by AUC) that was significantly greater in monkeys administered cidofovir alone (312% increase in males, 98% in females) than in those coadministered probenecid (32% increase in males, 3% in females). These results demonstrate that oral probenecid coadministration protects against the morphological evidence of nephrotoxicity and the accompanying decrease in renal clearance in monkeys receiving chronic intravenous cidofovir treatment. PMID- 9742651 TI - In vitro 2,3,7,8-tetrachlorodibenzo-p-dioxin interference with the anterior pituitary hormone adrenocorticortropin. AB - Treatment of male Sprague-Dawley rats with a single oral dose of 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) has been shown to increase serum adrenocorticotropin (ACTH) and decrease serum corticosterone. The present in vitro study was designed to assess whether TCDD has a direct effect on the anterior pituitary under basal and stimulated conditions. Primary anterior pituitary cell cultures were prepared from normal 180- to 220-g male Sprague Dawley rats and the cultures treated with 10(-9)-10(-19) M TCDD. Maximal secretion of ACTH occurred between 10(-11) and 10(-15) M TCDD for both medium (2 fold) and intracellular (1.5-fold) concentrations after 24 h TCDD exposure. TCDD treatment also caused an early (6 h) and persistent (10 days) increase in basal medium (1.4- to 2.8-fold) and intracellular (1.1- to 1.7-fold) ACTH concentrations. However, while stimulation with corticotropin-releasing hormone (CRH) increased intracellular ACTH 1.5- to 1.7-fold in pituitary cells treated for 24 h with 10(-9)-10(-13) M TCDD, ACTH secreted into the media was decreased by 30-50% compared with controls. Lastly, the secretagogue arginine-8-vaso pressin (AVP), did not increase the amount of ACTH secreted above levels observed with basal TCDD exposure. From this study, it appears that TCDD stimulates in vitro synthesis and secretion of ACTH by the anterior pituitary under basal conditions, but decreases the pituitary's responsiveness to CRH and AVP stimulation. PMID- 9742652 TI - 90-day feeding and one-generation reproduction study in Crl:CD BR rats with 17 beta-estradiol. AB - Over the past several years, there has been increasing concern that chemicals and pesticides found in the environment may mimic endogenous estrogens, potentially producing adverse effects in wildlife and human populations. Because estrogenicity is one of the primary concerns, a 90-day/one-generation reproduction study with 17 beta-estradiol was designed to set dose levels for future multigenerational reproduction and combined chronic toxicity/oncogenicity studies. The purpose of these studies is to evaluate the significance of a range of responses as well as to provide benchmark data for a risk assessment for chemicals with estrogen-like activities. This 90-day/one-generation reproduction study was conducted in male and female Crl:CD BR rats using dietary concentrations of 0, 0.05, 2.5, 10, and 50 ppm 17 beta-estradiol. Endpoints were chosen in order to evaluate both subchronic and reproductive toxicity. In addition, several mechanistic/biochemical endpoints were evaluated for their usefulness in follow-up studies. In the P1 generation, dietary administration of 2.5, 10, and 50 ppm 17 beta-estradiol produced dose-dependent decreases in body weight, body weight gain, food consumption, and food efficiency. At 10 and 50 ppm 17 beta-estradiol, minimal to mild nonregenerative anemia, lymphopenia, decreased serum cholesterol (50 ppm only), and altered splenic lymphocyte subtypes were also observed in the P1 generation. Additionally, at these concentrations, there were changes in the weights of several organs. Evidence of ovarian malfunction, characterized by reduced numbers of corpora lutea and large antral follicles, was observed at 2.5 ppm 17 beta-estradiol and above. Other pathologic changes in males and females fed 10 and 50 ppm 17 beta-estradiol included centrilobular hepatocellular hypertrophy; diffuse hyperplasia of the pituitary gland; feminization of the male mammary glands; mammary gland hyperplasia in females; increased number of cystic follicles in the ovary; hypertrophy of the endometrium and endometrial glands in the uterus; degeneration of seminiferous epithelium; and atrophy of the testes and the accessory sex glands. In the reproduction portion of this study, rats fed 10 or 50 ppm 17 beta-estradiol did not produce litters. While there was no evidence that the 50 ppm treated rats mated, 33.3% of the rats fed 10 ppm mated but did not produce litters. No effects on mating and fertility indices were observed in rats fed 0.05 and 2.5 ppm 17 beta-estradiol. Pup weights at birth were statistically decreased relative to control in the groups fed 0.05 and 2.5 ppm 17 beta-estradiol. Weights of the rats in the 0.05 ppm group recovered by postnatal day 4 and remained similar to control throughout the remainder of the study. The mean gestation length of the 0.05 ppm group was slightly, albeit not statistically significantly, shorter (0.5 days) than that of the control group, which may have contributed to the decrease in birth weight of the 0.05 ppm group. In contrast, the weights of the F1 generation rats fed 2.5 ppm 17 beta-estradiol remained decreased relative to the control group throughout the study. Parental administration of 17 beta-estradiol did not alter anogenital distance in male or female pups. The onset of sexual maturation, as measured by day of preputial separation in males and day of vaginal opening in females, was delayed in male rats fed 2.5 ppm (by 8.2 days) and was hastened in female rats fed 0.05 and 2.5 ppm (by 1.6 and 8.8 days, respectively). The age at vaginal opening ranged from 26 to 37, 26 to 35, and 21 to 25 days for rats fed 0, 0.05, and 2.5 ppm 17 beta-estradiol, respectively. Hence, the range of age at vaginal opening was similar between the control and 0.05 ppm group. The organ weight and pathologic alterations observed in the adult F1 generation rats were similar to those observed in the P1 generation rats. (ABSTRACT TRUNCATED) PMID- 9742653 TI - Effects of 17 beta-estradiol on serum hormone concentrations and estrous cycle in female Crl:CD BR rats: effects on parental and first generation rats. AB - The recently passed Food Quality Protection Act of 1996 requires the U.S. EPA to implement screening strategies for endocrine active compounds (EACs) within the next 2 years. Interpreting results from screening tests is complicated by the absence of traditional dietary rodent bioassay data with model estrogenic compounds such as 17 beta-estradiol. Thus, a 90-day/one-generation reproduction study with 17 beta-estradiol was designed to: (1) provide such baseline data; (2) set dose levels for multigeneration reproduction and combined chronic toxicity/oncogenicity studies; and (3) evaluate various mechanistic/biochemical endpoints for inclusion in these follow-up studies. The current article describes the effects of dietary administration of 0, 0.05, 2.5, 10, and 50 ppm 17 beta estradiol on the serum hormone concentrations and estrous cyclicity of female Crl:CD BR rats and evaluates a sampling strategy for measuring serum hormone levels in cycling female rats. Serum hormones were measured at three time points during a 90-day dietary exposure (1 week, 28 days, and 90 days) and in the F1 generation rats on postnatal day 98. Over the course of the 90-day feeding study for the P1 generation and from postnatal days 21 to 98 for the F1 generation, the estrous cycle was monitored daily in 10 rats/group. In P1 generation rats, dietary administration of 2.5, 10, and 50 ppm 17 beta-estradiol produced a dose dependent increase in serum estradiol (E2) concentrations at all time points. In contrast, administration of 0.05, 2.5, 10, and 50 ppm 17 beta-estradiol produced a dose-dependent decrease in serum progesterone (P4) concentrations on test day 90, which correlated with an absence of corpora lutea and ovarian atrophy. At 10 and 50 ppm 17 beta-estradiol, serum luteinizing hormone (LH) concentrations were consistently decreased at all time points and were decreased at 2.5 ppm on test day 90. Serum prolactin (PRL) concentrations were increased at 50 ppm 17 beta estradiol on test day 90. Serum follicle stimulating hormone (FSH) concentrations were either similar to the control levels or minimally changed at all time points. No F1 generation rats were produced at 10 or 50 ppm 17 beta-estradiol. In F1 generation rats, serum E2 concentrations were increased and P4 concentrations were decreased at a dietary concentration of 2.5 ppm 17 beta-estradiol, while serum concentrations of LH, FSH, and PRL were similar to the control. Dietary administration of 17 beta-estradiol at concentrations of 2.5 (both generations) and 10 and 50 ppm (P1 generation only) produced marked effects on the estrous cycle: decreased number of cycles, increased mean cycle length, and decreased number of normally cycling rats. The estrous cyclicity of rats fed 2.5 ppm 17 beta-estradiol appeared more severely affected in rats of the F1 generation than in rats of the P1 generation. Whether this increase in severity is related to an in utero exposure and/or greater mean daily intake of 17 beta-estradiol in the F1 generation rats in the postnatal period is unclear. Another goal of this study was to evaluate whether a single time point sampling strategy using cycling female rats could be used to detect compound-related changes in serum hormone concentrations. In evaluating a sampling strategy for measuring serum hormone levels, it appears that detection of compound-related alterations in serum hormone concentrations can be best detected by sampling during diestrus. Since the stage of the cycle dramatically influences hormone concentrations, large sample sizes (n = 50) are needed if serum hormone measurements are not matched with the stage of the cycle. The data indicate that this strategy of measuring serum hormone concentrations has utility in detecting compound-related effects within the confines of a traditional guideline study (subchronic, chronic, or multigenerational reproduction study). PMID- 9742654 TI - Effects of dietary 17 beta-estradiol exposure on serum hormone concentrations and testicular parameters in male Crl:CD BR rats. AB - A 90-day/one-generation reproduction study was conducted in male and female Crl:CD BR rats using dietary levels of 0, 0.05, 2.5, 10, and 50 ppm 17 beta estradiol. The goals of this study were to set dose levels and evaluate several mechanistic endpoints for inclusion in multigeneration reproduction and combined chronic toxicity/oncogenicity studies with 17 beta-estradiol. In this report we discuss the effects of dietary 17 beta-estradiol exposure on serum hormonal levels and sperm parameters from P1 and F1 male rats. Sperm parameters were also evaluated in recovery P1 and F1 male rats that were fed control diets for 105 and 103 days, respectively, following 97 and 86-94 days of estradiol exposure, respectively. Measurement of Sertoli cell number from F1 male rats was performed to test the hypothesis that in utero exposure to estrogens will decrease Sertoli cell number and sperm production. Other findings from this 90-day/one-generation reproduction study are summarized elsewhere. 17 beta-Estradiol produced a dose dependent decrease in body weight in P1 male rats at > or = 2.5 ppm and in the F1 male rats at 2.5 ppm. This decrease in body weight was due to a combination or reduced food consumption and food efficiency. In the recovery P1 males, body weight increased in the affected groups, albiet not to control levels, due to food consumption returning to control levels accompanied by an increase in food efficiency. However, in F1 males there was no corresponding rebound in body weight. In the P1 rats, exposure to 17 beta-estradiol decreased testis and epididymis weights in the 10 and 50 ppm groups, while no effects were seen in the P1 2.5 ppm group. In contrast, epididymis weights in the F1 and F1 recovery 2.5 ppm groups were statistically decreased; however, there were no histopathological effects observed. The decreases in testis weights in the P1 generation correlated with histopathologic evidence of interstitial cell atrophy and seminiferous tubule degeneration and reduced sperm production. Correlative changes in the epididymides of P1 rats were characterized by oligospermia or aspermia, the presence of germ cell debris in the lumen of tubules, and atrophy of epididymal tubules. 17 beta-Estradiol decreased testicular spermatid numbers, epididymal sperm numbers, and sperm motility in the P1 males in the 10 and 50 ppm groups, but not in the 2.5 ppm group. Following a 105-day recovery period in the P1 males, all sperm parameters and reproductive organ weights returned to control values except for the epididymal sperm count. Overall, the decline in testicular spermatid and epididymal sperm numbers in the P1 rats correlated with the reduced organ weights and the observed histopathological changes and appeared primarily related to the decrease in serum testosterone levels. In the F1 rats, no significant decreases were noted in the testicular spermatid number but a slight decrease in epididymal sperm number was seen in the 2.5 ppm group, which showed no evidence of recovery. Using morphometric analysis, no change was seen in the number of Sertoli cell nuclei per testis in F1 males. The pattern of hormonal responses seen in this study was characteristic of an estrogen receptor agonist such as 17 beta-estradiol: increased serum prolactin and decreased testosterone, luteinizing hormone, and follicle stimulating hormone levels. The data demonstrate that in utero and postnatal dietary administration of 17 beta estradiol at levels which increased serum estradiol levels to approximately 400% of control and decreased testosterone levels to 33% of control did not reduce the number of Sertoli cell nuclei per testis. PMID- 9742655 TI - Sensitivity of a Tier I screening battery compared to an in utero exposure for detecting the estrogen receptor agonist 17 beta-estradiol. AB - A Tier I screening battery for detecting endocrine active compounds (EACs) has been evaluated for its ability to identify 17 beta-estradiol, a pure estrogen receptor agonist. In addition, the responses obtained with the Tier I battery were compared to the responses obtained from F1 generation rats from a 90-day/one generation reproduction study with 17 beta-estradiol in order to characterize the sensitivity of the Tier I battery against the sensitivity of an in utero exposure for detecting EACs. The Tier I battery incorporates two short-term in vivo tests (5-day ovariectomized female battery; 15-day intact male battery) and an in vitro yeast transactivation system (YTS) for identifying compounds that alter endocrine homeostasis. The Tier I female battery consists of traditional uterotrophic endpoints coupled with biochemical and hormonal endpoints. It is designed to identify compounds that are estrogenic/antiestrogenic or modulate dopamine levels. The Tier I male battery consists of organ weights coupled with microscopic evaluations and a comprehensive hormonal assessment. It is designed to identify compounds that have the potential to act as agonists or antagonists to the estrogen, androgen, progesterone, or dopamine receptor; steroid biosynthesis inhibitors (aromatase, 5 alpha-reductase, and testosterone biosynthesis); or compounds that alter thyroid function. The YTS is designed to identify compounds that bind to steroid hormone receptors (estrogen, androgen, and progesterone) and activate gene transcription. The profile generated for 17 beta-estradiol was characteristic of the responses expected with a pure estrogen receptor agonist. In the female battery, responses to 17 beta-estradiol included increases in uterine fluid imbibition, uterine weight, estrus conversion, uterine stromal cell proliferation, uterine epithelial cell height, uterine progesterone receptor content, serum prolactin and estradiol levels, and decreases in uterine estrogen receptor content and follicle stimulating hormone and luteinizing hormone levels. In the male battery, responses to 17 beta-estradiol included decreases in absolute testis and epididymides weights, decreases in relative weights for androgen-dependent tissues (prostate, seminal vesicles, and accessory sex gland unit), hormonal alterations (decreased serum testosterone, dihydrotestosterone, and LH and increased serum prolactin levels), and microscopic alterations of the testis and epididymides. In the YTS for the estrogen receptor, 17 beta-estradiol had an EC50 value of 7.2 x 10(-9) M, while DHT and progesterone had little cross-activation. The androgen and progesterone receptor systems were less selective in that 17 beta-estradiol activated these systems within 3 orders of magnitude of the primary ligand. In the 90-day/one generation reproduction study, responses to dietary administration of 17 beta estradiol included alterations in organ weights, developmental landmarks, and hormonal levels. Comparison of the responses obtained with our Tier I battery and an in utero exposure demonstrates that the Tier I screening battery is as sensitive as an in utero exposure for detecting 17 beta-estradiol-induced alterations in hormonal homeostasis. PMID- 9742656 TI - Exposure to inorganic arsenic metabolites during early human development. AB - Because of the lack of data on the exposure to and toxic effects of inorganic arsenic during early human development, the transfer of arsenic to the fetus and suckling infant was studied in a native Andean population, living in the village San Antonio de los Cobres in the North west of Argentina, where the drinking water contains about 200 micrograms/liter. The concentration of arsenic in cord blood (median, 9 micrograms/liter) was almost as high as in maternal blood (median, 11 micrograms/liter), and there was a significant correlation between the two. Thus, at least in late gestation, arsenic is easily transferred to the fetus. The median concentration of arsenic in the placenta was 34 micrograms/kg, compared with 7 micrograms/kg previously reported for nonexposed women. Interestingly, essentially all arsenic in the blood plasma of both the newborns and their mothers was in the form of dimethylarsinic acid (DMA), the end product of inorganic arsenic metabolism. Similarly, about 90% of the arsenic in the urine of both the newborns and mothers in late gestation was present as DMA, compared with about 70% in nonpregnant women (p < 0.001). This may indicate that methylation of arsenic is increased during pregnancy and that DMA is the major form of arsenic transferred to the fetus. The increased methylation in late gestation was associated with lower arsenic concentrations in blood and higher concentrations in urine, compared with a few months postpartum. The arsenic concentrations in the urine of the infants decreased from about 80 micrograms/liter during the first 2 days of life to less than 30 micrograms/liter at 4.4 months (p = 0.025). This could be explained by the low concentrations of arsenic in the breast milk, about 3 micrograms/kg. PMID- 9742657 TI - Age-related increase in auditory impairment in monkeys exposed in utero plus postnatally to methylmercury. AB - Hearing impairment and deafness have been reported as a result of developmental and adult exposure to methylmercury; however, objective assessment of auditory function is generally lacking. This study extends previous research in our laboratory in which monkeys exposed to methylmercury from birth to adulthood exhibited high-frequency hearing impairment. Monkeys (Macaca fascicularis) were exposed throughout gestation and postnatally until 4 years of age to 0, 10, 25, or 50 micrograms/kg/day mercury as methylmercuric chloride. When they were 11 and 19 years of age, pure-tone detection thresholds for six frequencies between 0.125 and 31.5 kHz were determined by means of a psychophysical (behavioral) procedure. At 19 years of age, all five methylmercury-exposed monkeys exhibited elevated pure-tone thresholds compared with controls. Impairment was generally observed across the full range of frequencies. Comparisons of performance at 11 and 19 years revealed relatively greater deterioration in function in treated compared with control monkeys. These results extend previously reported evidence of deficits in auditory function produced by postnatal methylmercury exposure, and describe a pattern of deficit across frequencies different than that observed in the previous study. This study also provides evidence for development or acceleration of impairment of auditory function during aging as a consequence of developmental methylmercury exposure. PMID- 9742658 TI - Subchronic inhalation toxicity study of caprolactam (with a 4-week recovery) in the rat via whole-body exposures. AB - This study was designed to assess the potential subchronic inhalation toxicity of caprolactam when administered as a 3-micron aerosol from an aqueous solution to Sprague-Dawley CD rats (10/sex/group) via whole-body exposure. The study was enhanced with the inclusion of motor activity measurements and a functional observational battery to assess the neurotoxic potential of caprolactam. The rats were exposed at least 65 times over a 13-week period for 6 h per day, 5 days per week, to target concentrations (3 microns, mass median aerodynamic diameter) of 0, 25, 75, and 250 milligrams per cubic meter (mg/m3). An additional 10 animals/sex/group were similarly exposed and then held for a 4-week recovery period. Exposure levels were determined gravimetrically six times daily; one daily sample was analyzed by high-pressure liquid chromatography. No deaths were observed in the study during the exposure or recovery periods. Treatment-related responses such as labored breathing and nasal discharge were seen during many of the exposures. Similar responses as well as moist rales were seen during the nonexposure periods during the 13 weeks of exposure. However, these responses abated during the 4-week recovery period. There were no clearly treatment-related responses observed with ophthalmoscopic examinations, body weight measurements, food consumption measurements, neurobehavioral evaluations, clinical pathology evaluations, organ weight measurements, or macroscopic pathology examinations. Microscopic findings that were considered related to exposure to the test material were seen in the nasoturbinal tissues (hypertrophy/hyperplasia of goblet cells in the respiratory mucosa and intracytoplasmic eosinophilic material in epithelial cells of the olfactory mucosa) of the two higher-exposure group animals and in the laryngeal tissues (squamous/squamoid metaplasia/hyperplasia of the pseudostratified columnar epithelium covering the ventral seromucous gland) of all three exposure group animals. These changes were considered to be adaptive responses to an irritant (caprolactam). The keratinization of the metaplastic epithelium in the larynx was considered to be an adverse effect. By the end of the 4-week recovery period, there was complete regression of the keratinization in the larynx, but recovery of the adaptive nasoturbinal effects had not completely resolved. In conclusion, the whole-body exposure of Sprague-Dawley rats to caprolactam as a respirable aerosol for 6 h/day, 5 days/week, for 13 weeks at gravimetrically determined levels of 24, 70, and 243 mg/m3 resulted in respiratory tract effects (laryngeal) at the highest exposure level with complete recovery within 4 weeks postexposure. The results indicate that the no-observed adverse-effect level for caprolactam is 70 mg/m3, based on upper respiratory effects, with 243 mg/m3 representing a no-observed-effect level for systemic toxicity, neurotoxicity, and lower respiratory tract effects. PMID- 9742659 TI - Quantitative exposure of humans to an octamethylcyclotetrasiloxane (D4) vapor. AB - There is potential for human exposure to cyclic siloxanes by the respiratory route. To determine the pharmacokinetics of octamethylcyclotetrasiloxane (D4), a material commonly found in personal care products, the respiratory intake and uptake of D4 were measured in 12 healthy volunteers (25-49 years) on two occasions. Subjects inhaled 10 ppm D4 (122 micrograms/liter) or air (control) during a 1-h exposure via a mouthpiece in a double-blind, randomized fashion. Inspiratory and expiratory D4 concentrations were continuously measured. Exhaled air and plasma D4 levels were measured before, during, and after exposures. Individual D4 uptakes were measured under steady-state conditions during three rest periods (10, 20, and 10 min, respectively) alternating with two 10-min exercise periods. Mean D4 intake was 137 +/- 25 mg (SD) and the mean deposition efficiency was equivalent to 0.74/(1 + 0.45 VE), where VE is the minute ventilation. No changes in lung function were induced by the D4 vapor. Plasma measurements of D4 gave a mean peak value of 79 +/- 5 ng/g (SEM) and indicated a rapid nonlinear blood clearance. Using lung volume and respiratory surface area estimates based on functional residual capacity measurements, we developed a model and determined that the effective mass transfer coefficient for D4 was 5.7 x 10(-5) cm/s from lung air to blood. In an additional eight subjects, we compared D4 deposition with mouthpiece and nasal breathing at resting ventilations. For these individuals, mean deposition was similar for the two exposure protocols, averaging 12% after correction for exposure system losses. These are the first data describing the intake and absorption of D4 and they should contribute to a meaningful safety assessment of the compound. PMID- 9742661 TI - Dental embryology, anatomy, development, and aging. AB - Equine practitioners should be knowledgeable of dental anatomy and development to enhance their skill of age estimation of horses. The permanent teeth of horses are continually undergoing changes in shape and appearance. These changes may be used to suggest a reasonable age range for a horse. PMID- 9742660 TI - Acute respiratory exposure of human volunteers to octamethylcyclotetrasiloxane (D4): absence of immunological effects. AB - Humans are exposed to silicones in a number of commercial and consumer products. Some of these silicones, including octamethylcyclotetrasiloxane (D4), are volatile. Therefore, there is a potential for respiratory exposure. A pharmacokinetic analysis of respiratory exposure to D4 is presented in the accompanying paper (M. J. Utell et al., 1998, Toxicol. Sci. 44, 206-213). Possible immune effects of respiratory exposure to D4 are investigated in this paper. Normal volunteers were exposed to 10 ppm D4 or air for 1 h via a mouthpiece using a double-blind, crossover study design. Assays were chosen to screen for immunotoxicity or a systemic inflammatory response. Assessment of immunotoxicity included enumeration of peripheral lymphocyte subsets and functional assays using peripheral blood mononuclear cells. Because in humans there is no direct test for adjuvant effect of respiratory exposure, we analyzed proinflammatory cytokines and acute-phase reactants in peripheral blood, markers for a systemic inflammatory response, as surrogate markers for adjuvancy. These tests were repeated when the volunteers were reexposed to D4 approximately 3 months after this initial exposure. Blood was obtained prior to exposure, immediately postexposure, and 6 and 24 h postexposure. In these short-term, controlled human exposures, no immunotoxic or proinflammatory effects of respiratory exposure to D4 were found. PMID- 9742662 TI - Dental physical examination. AB - The objectives of the equine dental physical examination are to detect and quantify oral and dental disorders, to propose and carry out their treatment, and to implement management programs. The veterinarian should be able to offer a prognosis and to detail any future treatment or management plans that may be required. These objectives should take into account the cost of these procedures, and the veterinarian should be prepared to offer a cost-benefit analysis of the problem and the proposed cures. PMID- 9742663 TI - Dental imaging. AB - Radiography continues to be the initial test of choice in equine dental imaging for reasons of availability and ability to detect bone and tooth changes. Contrast radiography may be useful to characterize dental involvement in cases with draining tracts. For radiographically occult lesions, other modalities are useful. CT is better than plain radiography due to the inherent avoidance of superimposition of the opposite dental arcade, excellent bone density characterization, and good spatial resolution. Nuclear medicine may be useful to verify bone involvement in the dental region in cases in which the signs are particularly vague or not readily localized. Ultrasonography is an excellent test for soft tissue characterization and may assist with the characterization of suspected bone lysis, pathologic fractures, and abscesses. PMID- 9742664 TI - Congenital dental disease of horses. AB - Equine congenital dental deformities are not limited merely to those presented here; however, the examples discussed offer the reader an appreciation for the range of severity and complexity that may be found in affected horses. The veterinarian is obligated to provide the best possible care for the patient and to relieve animal suffering. The lack of definitive evidence for heritability of many of these defects can place the veterinarian in an untenable position, particularly when presented with literature that proclaims or suggests without evidence that a particular condition is inherited. In such cases, the veterinarian is encouraged to counsel owners, citing substantiated medical information, and to recommend that owners make the decision to eliminate the affected animals' ability to reproduce. PMID- 9742666 TI - Dental care and instrumentation. AB - Equine dentistry is not just carpentry work that involves floating the sharp enamel points off cheek teeth. Although floating is the most common and essential part of equine dentistry, every horse deserves a complete veterinary dental examination on a regular basis. Without such an examination, the equine practitioner can not determine the corrective procedures needed inside the horse's mouth. Dentistry for all ages is covered. Necessary instruments for a complete oral examination are also discussed. PMID- 9742665 TI - Pathophysiology of acquired dental diseases of the horse. AB - Periodontitis, infundibular necrosis, and periapical infection are dental diseases commonly affecting adult horses. Routine dental examinations and care may help to prevent these diseases. Further investigation of the treatment of horses with these diseases using local antimicrobial therapy, restorative dentistry, and endodontic therapy is needed. An understanding of the pathogenesis of these diseases aids in diagnosis and treatment. Gingival hyperplasia and odontogenic tumors are uncommon but should remain in a list of differential diagnoses when examining a horse with pertinent clinical signs. Recognition of odontogenic tumors as early as possible may facilitate surgery. Examination of the oral cavity of foals beyond the neonatal period should allow identification of brachygnathia and timely treatment when indicated. PMID- 9742667 TI - Traumatic dental disease and soft tissue injuries of the oral cavity. AB - Trauma to the oral cavity can result in an array of injuries affecting teeth, bone, and soft tissue. A thorough examination of the oral cavity is often facilitated by employing a full-mouth speculum after the horse has been tranquilized. Identification of broken, loose, or split teeth; fractures of the premaxilla or mandible; and avulsion or laceration of soft tissue structures such as the lips or tongue is usually straightforward. Treatment options vary depending on the structure involved; however, appropriate treatment generally results in a functional and cosmetically acceptable end result. PMID- 9742668 TI - Dental sepsis. AB - Dental sepsis or periapical abscess formation constitutes a large percentage of dental conditions that afflict horses. Dental sepsis occurs when the pulp chamber of the tooth is exposed to the oral cavity or external environment, allowing bacterial localization with resulting infection. Although acute, primary, septic pulpitis in horses is rare, dental sepsis often results from colonization of the pulp chamber with pathogenic bacteria secondary to maleruption or impaction of teeth with secondary alveolar bone lysis, primary fractures of the tooth, mandible, or maxilla, periodontal disease, or infundibular necrosis. The sequela to pulpal infection are extensions into the periradicular tissues and mandibular or maxillary periapical abscess formation. PMID- 9742669 TI - Dental disease in geriatric horses. AB - The dental management of geriatric horses can be a rewarding challenge to the practitioner. Owners become dissatisfied when their expectations are unrealistic. Consequently, communication between the owner and the practitioner is essential prior to the start of any dental procedure in a geriatric horse. Owners often expect the practitioner to correct what has been neglected for years. It is critical that the owner understand the possible complications associated with dental procedures and that some procedures (e.g., trephination) may necessitate protracted care. Often, when a tooth has been removed, there is a need for more frequent masticatory examinations to curtail any potential problems (i.e., development of step mouth). The owner needs to be aware of the extra dental maintenance costs that must be included in the upkeep of the horse. PMID- 9742670 TI - Dental surgery in horses. AB - Dental surgery is most often directed at removal of diseased or injured teeth by the least invasive method possible. Some procedures available can preserve traumatized or infected teeth. Complications of dental surgery are well documented and often encountered by veterinary surgeons. Principles of debridement, curettage, lavage and ventral drainage combined with appropriate medicinal management can reduce complications and lead to successful outcome after dental surgery. PMID- 9742671 TI - Complications of dental surgery. AB - Both retrospective data and clinical experience indicate that complications of dental surgery are occasionally encountered and, to some extent, are inevitable. Many of the reported complications related to dental surgery such as incomplete removal of diseased teeth or removal of the wrong tooth can be avoided with sound preoperative planning and intraoperative technique. Diseased teeth should be properly identified prior to and during surgery. In addition, complete removal of the diseased tooth must be performed. Use of intraoperative radiographic examination to confirm the location of the diseased tooth and to document its removal cannot be overemphasized. Iatrogenic fracture of the maxillary or mandibular alveolar walls or palatine bone can be avoided by proper placement of the dental punch. The chances of developing incisional drainage or secondary sinusitis can be reduced by use of appropriate systemic antibiotics. These factors should guide the surgical approach to dental surgery to reduce the likelihood of developing common complications. PMID- 9742672 TI - Dental corrective procedures. AB - This article explains what is needed for successful extraction of diseased cheek teeth and how to realign the occlusal surface. Incisor teeth procedures and correcting abnormalities of cheek tooth crown wear are also discussed along with wolf and floating teeth. PMID- 9742673 TI - Activity-dependent modulation of adaptation produces a constant burst proportion in a model of the lamprey spinal locomotor generator. AB - The neuronal network underlying lamprey swimming has stimulated extensive modelling on different levels of abstraction. The lamprey swims with a burst frequency ranging from 0.3 to 8-10 Hz with a rostrocaudal lag between bursts in each segment along the spinal cord. The swimming motor pattern is characterized by a burst proportion that is independent of burst frequency and lasts around 30% 40% of the cycle duration. This also applies in preparations in which the reciprocal inhibition in the spinal cord between the left and right side is blocked. A network of coupled excitatory neurons producing hemisegmental oscillations may form the basis of the lamprey central pattern generator (CPG). Here we explored how such networks, in principle, could produce a large frequency range with a constant burst proportion. The computer simulations of the lamprey CPG use simplified, graded output units that could represent populations of neurons and that exhibit adaptation. We investigated the effect of an active modulation of the degree of adaptation of the CPG units to accomplish a constant burst proportion over the whole frequency range when, in addition, each hemisegment is assumed to be self-oscillatory. The degree of adaptation is increased with the degree of stimulation of the network. This will make the bursts terminate earlier at higher burst rates, allowing for a constant burst proportion. Without modulated adaptation the network operates in a limited range of swimming frequencies due to a progressive increase of burst duration with increasing background stimulation. By introducing a modulation of the adaptation, a broad burst frequency range can be produced. The reciprocal inhibition is thus not the primary burst terminating factor, as in many CPG models, and it is mainly responsible for producing alternation between the left and right sides. The results are compared with the Morris-Lecar oscillator model with parameters set to produce a type A and type B oscillator, in which the burst durations stay constant or increase, respectively, when the background stimulation is increased. Here as well, burst duration can be controlled by modulation of the slow variable in a similar way as above. When oscillatory hemisegmental networks are coupled together in a chain a phase lag is produced. The production of a phase lag in chains of such oscillators is compared with chains of Morris-Lecar relaxation oscillators. Models relating to the intact versus isolated spinal cord preparation are discussed, as well as the role of descending inhibition. PMID- 9742674 TI - Dynamics extraction in multivariate biomedical time series. AB - A nonlinear analysis of the underlying dynamics of a biomedical time series is proposed by means of a multi-dimensional testing of nonlinear Markovian hypotheses in the observed time series. The observed dynamics of the original N dimensional biomedical time series is tested against a hierarchy of null hypotheses corresponding to N-dimensional nonlinear Markov processes of increasing order, whose conditional probability densities are estimated using neural networks. For each of the N time series, a measure based on higher order cumulants quantifies the independence between the past of the N-dimensional time series, and its value r steps ahead. This cumulant-based measure is used as a discriminating statistic for testing the null hypotheses. Experiments performed on artificial and real world examples, including autoregressive models, noisy chaos, and nonchaotic nonlinear processes, show the effectiveness of the proposed approach in modeling multivariate systems, predicting multidimensional time series, and characterizing the structure of biological systems. Electroencephalogram (EEG) time series and heart rate variability trends are tested as biomedical signal examples. PMID- 9742675 TI - Sensitivity analysis of a model of mammalian neural membrane. AB - The sensitivity of the strength-duration (S-D) relationship to changes in the parameters describing the sodium channel of mammalian neuronal membrane was determined by computer simulation. A space-clamped patch of neuronal membrane was modeled by a parallel nonlinear sodium conductance, linear leakage conductance, and membrane capacitance. Each parameter that governs the activation (m) and inactivation (h) variables of the sodium channel was varied from -50% to +50% of its default value, and for each variation a S-D relationship was generated. Individual changes in six of the eleven parameters (alpha mA, alpha mD, alpha hA, beta mA, beta mB, and beta hB) generated substantial changes in the rheobase current and chronaxie time (Tch) of the model. Changing the parameter values individually did not correct for the model's failure to generate excitation after the release from a long duration hyperpolarization (anode break excitation). Scaling a combination of five parameters (alpha mA, alpha mB, alpha hA, beta mA, and beta hB) by an equal amount produced a model that generated anode break excitation and increased Tch, but also decreased the amplitude of the action potential. To reproduce the amplitude of the action potential, the maximum sodium conductance and sodium Nernst potential were increased. These modifications generated a model that had S-D properties closer to experimental results, could produce anode break excitation, and reproduced the action potential amplitude. PMID- 9742676 TI - Timing of secondary vestibular neuron responses to a range of rotational head movements. AB - Secondary vestibular neurons exhibit a wide variety of responses to a head movement, with the response of each secondary neuron depending upon the particular primary afferents converging onto it. A single head movement is thereby registered in a distributed manner. This paper focuses on implications of afferent convergence to the relative timing of secondary neuron response modulation during rotational movements about a combination of horizontal axes. In particular, the neurons of interest are those that receive input from afferents innervating the vertical semicircular canals, and the movements of interest are those that have a sinusoidal component about one vertical canal axis and a sinusoidal component about another, approximately orthogonal, vertical canal axis. Under these conditions, the present research shows that it is possible for two or more secondary neurons to have a different relative timing of response (i.e., different relative phase of the periodic modulation in firing rate) for different head movements, and for the neurons to switch their order of response for different movements. For particular head movements, those same neurons will respond in phase. From the point of view of the nervous system, the relative timing of neuron responses may tell which movement is taking place, but with certain restrictions as discussed in the present paper. Shown here is that, among those head movements for which the two components of rotation may be at any phase relative to one another and have any relative amplitude, an in-phase response of just two neurons cannot identify a single motion. Two neurons that respond in phase for one motion must respond in phase for an entire range of motions; all motions in that range are thus response-equivalent, in the sense that the pair of neurons cannot distinguish between the two motions. On the other hand, an in phase response of three neurons can identify a single motion, for certain patterns of primary afferent convergence. PMID- 9742677 TI - Individual differences in brain dynamics: important implications for the calculation of event-related band power. AB - Measures of event-related band power such as event-related desynchronization (ERD) are conventionally analyzed within fixed frequency bands, although it is known that EEG frequency varies as a function of a variety of factors. The question of how to determine these frequency bands for ERD analyses is discussed and a new method is proposed. The rationale of this new method is to adjust the frequency bands to the individual alpha frequency (IAF) for each subject and to determine the bandwidth for the alpha and theta bands as a percentage of IAF. As an example, if IAF equals 12 Hz, the widths of the alpha and theta bands are larger as compared to a subject with an IAF of, e.g., only 8 Hz. The results of an oddball paradigm show that the proposed method is superior to methods that are based on fixed frequencies and fixed bandwidths. PMID- 9742678 TI - Role of temperature in quanta mechanisms of facilitation in the frog neuromuscular junction. AB - The results of computer simulations on the Double Barrier Synapse (DBS) model are presented which quantify the relationship between the synapse parameters and the quanta transfer process. The DBS model is applicable to a variety of states of synaptic activity, and by changing the synapse parameters it is possible to simulate various conditions of quanta transmission. The influence of the bathing solution temperature change on the synaptic parameters under different conditions of transmitter release in the frog neuromuscular junction is investigated. Simulations demonstrate that several synaptic parameters, including the parameters of the presynaptic membrane, are not affected by the temperature change. It is shown that a stimulation frequency exists at which the steady-state level of facilitation during a long train of stimuli is the same for a wide range of temperatures. PMID- 9742679 TI - A single pacemaker cell model based on the phase response curve. AB - A single pacemaker cell model and its response to repetitive external depolarization stimulations is described in this paper. This model is a simple model based on the two most important functional properties of the cardiac pacemaker cells. The first property is the intrinsic pacemaker cycle length, which is an 'internal' parameter of the cell, describing the most important feature of a pacemaker cell. The second functional property is the phase response curve (PRC), which is an 'overall collective' function: it contains all the 'information' about the possible interactions of the pacemaker cell with the outside world (external stimulus, interaction with surrounding cells, etc.). This study demonstrates that by representing the pacemaker cell only by two fundamental features, and by applying a simple physical-mathematical model, a global picture of the system can be achieved, allowing us to explore qualitatively various physiological phenomena related to the pacemaker function. For example, we demonstrated that the PRC is a crucial parameter in the prediction of the entrainment phenomena of a single pacemaker cell in response to a periodic train of depolarization pulses. Specifically, the PRC permits a quantitative determination of the 1:1 synchronization range for a single pacemaker cell and an external depolarization pulse. Moreover, we show that the PRC can be used to represent the type of external stimulus applied to the pacemaker (e.g. depolarization pulse) and its intensity. Therefore, the PRC emerges as an important determinant and a useful 'tool' for the understanding of the dynamic interaction of pacemaker cells with the outside world. As a result of our simulations, we unveil a new important parameter: the 'degree of influence', which determines the range of 1:1 synchronization between an external depolarization pulse and a pacemaker cell. This interaction parameter is a direct function of the PRC parameters. It appears to be a helpful 'tool' for the understanding of synchronization and mutual entrainment mechanisms between the pacemaker cell and an external stimulus, and therefore it supports the basic importance of the PRC in the description and determination of these mechanisms. PMID- 9742680 TI - A pacemaker cell pair model based on the phase response curve. AB - A pacemaker cell pair model and the dynamic interaction between the two pacemaker cells is described in this paper. It is an extension of our single pacemaker cell model, in which we studied its response to repetitive external depolarization stimulations. This model is a simple model based on the two most important functional properties of the cardiac pacemaker cells: its intrinsic pacemaker cycle length, which is an 'internal' parameter of the cell, and the phase response curve (PRC), which is an 'overall collective' function. The PRC contains all the 'information' about the possible interactions of the pacemaker cell with the outside world (interaction with surrounding cells, external stimulus, etc.). First, we examined the properties and solutions of 1:1 synchronization between two pacemaker cells. We found that in order to achieve synchronization between two pacemaker cells, there should be limitations on the PRC parameters, which depend on the cells intrinsic cycle lengths. Next, we investigated the 2:1 entrainment state between two interacting pacemaker cells. We found that there is not necessarily a unique solution for this state as there was for the 1:1 state. Finally, we ran our computer model to investigate the properties of more complex patterns of entrainment between two pacemaker cells. As a result of our analytical study, we unveil two new important parameters, which are fully defined as a function of the PRC parameters: (1) the 'accelerator factor' which describes the tendency of a pair of interacting pacemaker cells to synchronize at a common cycle length, which is closer to the faster cycle of the pair; (2) the 'degree of coupling', which describes the range of the 1:1 synchronization and the 'strength' of the interaction between a pair of interacting pacemaker cells. Those two interaction parameters arise as helpful 'tools' for the understanding of synchronization and mutual entrainment mechanisms between pacemaker cells. Therefore, this study establishes the PRC as an important determinant and a useful approach for the understanding of the dynamic interaction of pacemaker cells among themselves and with the outside world. PMID- 9742681 TI - Thermal stability and reversibility of secondary conformation of alpha-crystallin membrane during repeated heating processes. AB - Reflectance FT-IR/DSC microspectroscopy was first used to study the structural conformation of alpha-crystallin membranes in the heating-cooling-reheating cycle. The thermotropic transition and the changes in secondary structure of alpha-crystallin membrane during heating and reheating processes were investigated. A thermal transition ranging between 50 and 70 degrees C with a midpoint at 60 degrees C for the alpha-crystallin membrane was easily obtained from the three-dimensional plots of the reflectance FT-IR spectra as a function of temperature. The secondary structural components of the alpha-crystallin membrane were modified step-by-step with the increase of temperature from 25 to 120 degrees C, but restored to original values after cooling to 25 degrees C. During the heating process, the compositions of the alpha-helix, random coil and beta-sheet structure decreased with temperature, but the content of the beta-turn structure increased, however, all of them were restored after cooling. The absence of significant alteration in the secondary structures for the alpha crystallin membrane before and after the first-heating process strongly suggests the high thermal stability and reversibility of alpha-crystallin. Interestingly, the thermal behavior of the first-heated alpha-crystallin membrane during the reheating process exhibited a unique thermal behavior with two transitional temperatures at 35-50 and 55-70 degrees C. The reflectance FT-IR/DSC microscopic data indicated that alpha-crystallin in the membrane state had higher thermal stability and reversibility. PMID- 9742682 TI - Solvent accessibility studies on glycosaminoglycans. AB - Theoretical calculations on the solvent accessible surface area of the commonly occurring glycosaminoglycans (GAGs) have been carried out by employing the solvent accessibility technique of Lee and Richards. The results indicate that the average variation of solvent accessible surface area (ASA) for the different atoms is in the range of 2-28 A2. The average ASA for carbon (9.02 A2) and oxygen (19.3 A2) are relatively very high compared to that for nitrogen and sulfur. The total contribution of ASA by the buried atoms in all the molecules has been found to be small. Heparin complexed with the protein is less accessible to the solvent than the uncomplexed. The iduronic acid is also found to have a varied ASA due to its conformational flexibility. The sulfate groups in heparin have 50% of the total ASA of the molecule. Residue analysis indicates that in general D configuration residues have higher ASA than L-configuration residues. PMID- 9742683 TI - Study of alpha-helix to beta-strand to beta-sheet transitions in amyloid: the role of segregated hydrophobic beta-strands. AB - A conformational analysis including three polypeptide chains known to be amyloidogenic and neurotoxic has shown the occurrence of low probability hydrophobic beta-strands stabilized by intramolecular interactions. It is argued that by engaging in non-bonded and hydrophobic interactions these beta-strands seed the assembly of beta-sheets in amyloid fibrils following a non-cooperative mechanism dissimilar to beta-sheet folding in proteins. Molecular models of amyloid fibrils formed by such beta-strand templates were built. It is shown that the parallel alignment of beta-strands creates an extensive hydrophobic surface whereas the antiparallel alignment causes the formation of hydrophobic and hydrophilic aggregates. A comparison with experimental data and previous calculations is established. PMID- 9742684 TI - Carboxyl groups at the membrane interface as molecular targets for local anesthetics. AB - The interaction of the tertiary amine drugs chlorpromazine and dibucaine in their cationic form with carboxyl groups at the membrane surface is studied at concentrations relevant to anesthesia. Spin-labeled stearic acid is used both to provide the carboxyl groups and to monitor binding and ionization behavior in egg lecithin liposomes. Membrane anesthetic concentrations are spectrophotometrically obtained. They are shown to determine the drug influence on carboxyl groups at the membrane surface, independently of aqueous concentrations. The intramembrane association constants (related to the usual aqueous phase ones through the partition coefficient) of the drugs with fatty acids are determined. The same value (10(2) M-1) is obtained for both drugs, suggesting that it is approximately the same for all tertiary amine local anesthetics. pH titrations of anesthetic treated spin-labeled membranes are performed. The observed shifts in the fatty acid pK are higher than can be produced assuming uniform distribution of the drug in the membrane surface, implying that there is an increased affinity of local anesthetics for superficial carboxyl. This affinity could account for the resting block of voltage-gated Na+ channels. Under these considerations, local anesthetic binding sites at voltage-gated Na+ channels and at sarcoplasmic reticulum Ca(2+) ATPase are proposed. PMID- 9742685 TI - Retinol and retinoic acid bind to a surface cleft in bovine beta-lactoglobulin: a method of binding site determination using fluorescence resonance energy transfer. AB - Two potential ligand binding sites in the lipocalin beta-lactoglobulin have been postulated for small hydrophobic molecules such as retinol or retinoic acid. An agreement on one of the two alternatives, an interior cavity or a surface cleft, however, has not been achieved. In order to discriminate between these two possibilities, we measured the efficiency of fluorescence resonance energy transfer between the two intrinsic Trp-residues of beta-lactoglobulin and the ligands retinol, retinoic acid and bis-ANS. Using the crystallographic coordinates of beta-lactoglobulin, this efficiency could be accurately computed for both the interior cavity and the surface cleft as ligand binding sites. For the surface cleft, the theoretical value was found to be in excellent agreement with the measured value, whereas for the interior cavity any reasonable agreement would require a dramatic ligand-induced conformational change that can be ruled out due to the protein's known structural stability. Our conclusion that these ligands bind to the surface pocket rather than the interior cavity was further confirmed by competitive binding studies. PMID- 9742686 TI - The entropy generation in visual-pigment system by the absorption of light. AB - We first showed a general theory that reception of information from the outside in a receptor system is accompanied both by an inflow of entropy and by a generation of entropy depending on the reliability of the actual reception mechanism. Then, considering a case for the absorption of light by the visual pigment system of visual cell, we calculated the time(t)-dependent change in the number N2(t) of excited visual-pigments to obtain the entropy increase. We thus arrived at the following four conclusions: (1) One or two photons can be detected with a reliability of at least 54%; (2) In compensation for this detection, entropy > 1.12 x log2 is generated; (3) An incident photon of frequency v from a ligh source of temperature Ts yields an entropy of hv/Ts; and (4) Depending on the characteristics of the visual-pigment system, another entropy being different from (2) is generated in proportion to N2(t). PMID- 9742687 TI - X-ray diffraction studies on the structural changes of rigor muscles induced by binding of phosphate analogs in the presence of MgADP. AB - To clarify the structure of the ATP hydrolysis intermediates (ADP.Pi bound state) formed by actomyosin crossbridges, the effects of various phosphate analogs in the presence of MgADP on the structures of the thin and thick filaments in glycerinated rabbit psoas muscle fibers in the rigor state have been investigated by X-ray diffraction with a short exposure time using synchrotron radiation. When MgADP and phosphate analogs such as metallofluorides (BeFx = 3,4 and AlF4) and vanadate (VO4(Vi)) were added to rigor fibers in the presence of the ATP depletion backup system, the intensities of the actin-based layer lines were markedly weakened. The greatest effect (approximately 50% decrease in intensity) was observed in the presence of BeFx among the analogs examined. The intensity distribution of the 5.9 nm actin-based layer line shifted towards that observed in the Ca(2+)-activated fibers, while the first actin layer line at approximately 1/36.7 nm-1 retained a rigor-like profile with an intensity weakened by approximately 50%. The intensity of the equatorial 10 reflection increased while that of the 11 reflection changed little, resulting in only a small increase (approximately 1.7 fold) in the intensity ratio of the 10 to the 11 reflection. No resting-like pattern appeared upon the addition of MgADP and BeFx. These results indicate that a substantial fraction (approximately 40%) of the myosin heads dissociate from actin but the detached heads remain in the vicinity of the actin filaments when MgADP and BeFx bind. The states produced by binding phosphate analogs to a rigor muscle differ from the resting-like state produced by adding them to a contracting muscle (Takemori et al., J. Biochem. (Tokyo) 117 (1995) 603-608). Our conclusion put forward to explain the data is that one of the two heads of a crossbridge is detached and the other retains a rigor-like attachment. PMID- 9742688 TI - Binding of modified fragments of the Shigella dysenteriae type 1 O-specific polysaccharide to monoclonal IgM 3707 E9 and docking of the immunodeterminant to its modeled Fv. AB - The O-specific polysaccharide (O-SP) of Shigella dysenteriae type 1 has been shown by others to have the structure-->3)-alpha-L-Rhap-(1-->3)-alpha-L-Rhap-(1- >2)-alp ha-D- Galp-(1-->3)-alpha-D-GlcpNAc-(1-->. We have shown in the past that IgM 3707 E9, an anti S. dysenteriae type 1 O-SP monoclonal antibody, binds specifically to the -alpha-L-Rhap-(1-->2)-alpha-D-Galp-determinant of the polysaccharide. In this report we show that determinant to have hydrogen bonds, necessary for binding to the antibody, involving positions 3, 4 and 6 of the galactopyranosyl residue. The hydroxyl groups of the rhamnopyranosyl moiety of the immunodeterminant appear not to partake in hydrogen-bond interactions with the antibody. A model is presented of the Fv of IgM 3707 E9 based on our previously established cDNA-sequence and two known, highly homologous immunoglobulin crystal structures. The methyl glycoside of the immunodeterminant alpha-L-rhamnopyranosyl-(1-->2)-alpha-D-galactopyranose is docked to the combining area of the Fv. PMID- 9742689 TI - Conversion of N-sulfated glucosamine to N-sulfated mannosamine in an unsaturated heparin disaccharide by non-enzymatic, base-catalyzed C-2 epimerization during enzymatic oligosaccharide preparation. AB - A novel disaccharide was isolated beside the predominant trisulfated disaccharide, delta HexA(2-O-sulfate)(alpha 1-4)GlcN(2-N-,6-O-disulfate) (delta HexA and GlcN represent 4-deoxy-alpha-L-threo-hex-4-enepyranosyluronic acid and D glucosamine, respectively) after treatment of porcine intestinal heparin with Flavobacterium heparinase. It accounted for 18% of total disaccharides. The structure was characterized by secondary ion mass spectrometry, enzymatic digestions, amino sugar analysis, and 500 MHz one- and two-dimensional 1H NMR spectroscopy as delta HexA(2-O-sulfate) (alpha 1-4)ManN(2-N-,6-O-disulfate), where ManN represents D-mannosamine. The C-2 epimerization from delta HexA(2-O sulfate) (alpha 1-4)GlcN(2-N-,6-O-disulfate) to delta HexA(2-O-sulfate) (alpha 1 4)ManN(2-N-,6-O-disulfate) was also demonstrated to take place in vitro under very mild alkaline conditions. Hence, the latter compound is not a biosynthetic product, but is most likely an artifact generated by non-enzymatic, base catalyzed C-2 epimerization during enzymatic preparation of heparin oligosaccharides. The present results warn that the formation of the C-2 epimerized compound has to be circumvented in the structural analysis of heparin/heparan sulfate. PMID- 9742690 TI - Solid-phase synthesis of the B-chain of human alpha 2HS glycoprotein. AB - The B-chain of human alpha 2HS glycoprotein 1, a heptacosapeptide carrying a trisaccharide (sialyl T) side chain, was synthesized. Prior to the Fmoc-based solid-phase synthesis of the glycopeptide, the benzyl-protected glycosyl serine building block 6 was prepared via beta-stereoselective glycosylation of the 2 azido-2-deoxygalactosyl serine 11 with the sialyl galactosyl trichloroacetimidate 9. An automated peptide synthesizer was efficiently used for the elongation of the entire peptide chain except for the coupling with 6. The synthesized glycopeptide was cleaved from the resin by the TFA method. The resultant mixture of the benzylated glycopeptides was treated with TMSOTf-thioanisole in TFA and then with aq NaHCO3 and 1,4-dithiothreitol to give 1. PMID- 9742691 TI - Oligosaccharide fragments of the type III group B streptococcal polysaccharide derived from S. pneumoniae type 14 capsular polysaccharide by a chemoenzymatic method. AB - Partial N-deacetylation fo the GlcNAc residues in S. pneumoniae type 14 capsular polysaccharide (Pn14-PS) backbone was achieved by treatment with base, and the product was subsequently enzymatically sialylated at the 3-O-positions of the terminal galactose residues. The resultant, partially N-deacetylated type III Group B streptococcus capsular polysaccharide (GBSIII-PS) was subjected to nitrous acid deamination, which resulted in the degradation of GBSIII-PS polysaccharide into oligosaccharides containing increasing numbers of the identical repeating units. The oligosaccharides were then separated by passage through a Superdex 30 column and characterized by ESIMS and NMR spectroscopic analysis. PMID- 9742692 TI - Extracellular polysaccharides and polysaccharide-containing biopolymers from Azospirillum species: properties and the possible role in interaction with plant roots. AB - This paper reviews the results obtained in studies of the extracellular polysaccharides, lipopolysaccharide-protein complexes, polysaccharide-lipid complexes, lipopolysaccharides, and O-specific polysaccharides from bacteria of the genus Azospirillum. On the basis of present knowledge, the possible roles of the extracellular polysaccharides and polysaccharide-containing complexes of azospirilla in interaction with the roots of plants are discussed. Some pieces of evidence are considered in light of the lectin hypothesis originally proposed for the legume-Rhizobium symbiosis. In the context of these views of Azospirillumcereal associative pairs, a key process at the early stages of the interaction is the specific reaction of cereal root lectins with the extracellular polysaccharide components, containing N-acetyl-D-glucosamine as part of their structure. PMID- 9742693 TI - A second European collaborative study on polymerase chain reaction for Toxoplasma gondii, involving 15 teams. AB - In order to investigate the accuracy and practicability of the polymerase chain reaction (PCR) in the antenatal diagnosis of congenital toxoplasmosis, a collaborative study involving 15 European laboratories was performed under the auspices of the Biomed 2 Programme of the European Community. Each team received 12 aliquots (four negative, eight positive) of 'artificial samples' made of amniotic fluid spiked with tachyzoites of the RH strain of Toxoplasma gondii. Each team performed its own PCR protocol (all were different). Nine of the 15 laboratories were able to detect a single parasite, but two of the 15 found all samples negative. Four of the 15 laboratories found one or more control samples to be falsely positive. This study highlights the lack of homogeneity between PCR protocols and performance and underlines the need for an external quality assurance scheme which could provide 'reference' samples that could be used by any laboratory wanting to establish and maintain an accurate diagnostic test based on PCR. PMID- 9742694 TI - Cloning, sequencing and expression of the flagellin core protein and other genes encoding structural proteins of the Vibrio cholerae flagellum. AB - Vibrio cholerae is a Gram-negative bacterium with a single polar flagellum. Motility is an important virulence factor for this non-invasive pathogen. We cloned and sequenced a locus in V. cholerae V86 (El Tor, Inaba) that contained five different structural genes of the flagellum. The cloned genes and their products were assigned names and functions based on homology with sequences of similar genes and their products from other related bacteria. All of these genes of V. cholerae V86, namely, flgI, J, M, L and flaA, were transcribed in the same direction. These genes respectively encoded the P- and L-ring proteins, the hook associated proteins 1 and 3 and the flagellin core protein of the flagellum. Our data indicated the presence of more than one flagellar locus in V. cholerae which could provide a means of immunoavoidance during infection. When compared with homologs in other bacteria, the flagellin core protein of V. cholerae exhibited conservation in the N- and C-termini, but had diverged in the central region. PMID- 9742695 TI - Homologous transformation of Trichoderma hamatum with an endochitinase encoding gene, resulting in increased levels of chitinase activity. AB - A 42-kDa endochitinase encoding gene, Tham-ch, was cloned by screening the genomic library of Trichoderma hamatum strain Tam-61 with a PCR-amplified chitinase sequence from the same fungus. Tham-ch with its own regulatory sequences was reintroduced into the host strain. The integration of the transforming construct was stable only in one copy. Homologous integration occurred in nine transformants, while non-homologous integration was detected in one transformant. All but one transformant expressed higher levels of chitinase activity in comparison to the wild-type recipient strain; the maximum level of increase was 5-fold. Duplicating the copy number of the highly conserved approximately 42-kDa endochitinase encoding gene appears to be one potential means by which the biocontrol capability of the Trichoderma species might be improved. PMID- 9742696 TI - Tn5563, a transposon encoding putative mercuric ion transport proteins located on plasmid pRA2 of Pseudomonas alcaligenes. AB - Sequence analysis of pRA2, an endogenous 33-kb plasmid from Pseudomonas alcaligenes NCIB 9867 (strain P25X), revealed the presence of a 6256-bp transposon of the Tn3 family, designated Tn5563. Tn5563, which is flanked by two 39-bp inverted repeats, encodes a transposase, a resolvase, and two open reading frames which share amino acid sequence similarities with the mercuric ion transport proteins MerT and MerP encoded by several mer operons. However, no other mer operon genes were found on Tn5563. Sequencing of a RP4::XIn hybrid plasmid indicates possible interactions between pRA2 and the P25X chromosome mediated by Tn5563. PMID- 9742698 TI - Isolation and characterization of a chitinase cDNA from the entomopathogenic fungus, Metarhizium anisopliae. AB - A novel chitinase gene was isolated from Metarhizium anisopliae grown in a medium containing chitin as the sole carbon source. Comparisons of nucleotide sequence of the isolated gene with those of other fungal chitinase genes showed low sequence identity (24.4-36.4%) except for the active site of chitinase. In addition, molecular mass determination of the fused gene product separated on a gel showed that the fused chitinase seems to be about 70 kDa, while the molecular mass calculated from the deduced amino acid sequence can be about 58 kDa. These molecular masses were different from values of 33 kDa for an endochitinase and 110 kDa for an exochitinase (N-acetylglucosaminidase) from M. anisopliae published previously. PMID- 9742697 TI - Antibiotic stress induces a large amount of outer membrane protein in Pseudomonas aeruginosa. AB - To investigate the bacterial response to antibiotic stress, we analyzed the outer membrane proteins of Pseudomonas aeruginosa grown in the presence of a sub minimum inhibitory concentration of antibiotics. Among the antibiotics tested, fluoroquinolones and streptonigrin induced a large amount of outer membrane protein with a molecular mass of 43 kDa. This protein is most likely the stress responsive protein, since the quinolone-resistant mutants with a higher minimum inhibitory concentration of antibiotic than the wild-type strain produced a large amount of 43-kDa protein only in the presence of sub-minimum inhibitory concentration of the mutants itself, but not that of the antibiotic-susceptible wild-type strain. The sequence of N-terminal 15 amino acids of the 43-kDa protein was identical to that of pyocin R1. However, purified pyocin R1 failed to accumulate in the outer membrane. Thus, we concluded that the 43-kDa protein (pyocin R1) is the antibiotic-stress-induced outer membrane protein. PMID- 9742699 TI - PKC1, encoding a protein kinase C, and FAT1, encoding a fatty acid transporter protein, are neighbors in Cochliobolus heterostrophus. AB - A protein kinase C gene (PKC1) and adjacent DNA of the filamentous ascomycete Cochliobolus heterostrophus was cloned and sequenced. The deduced amino acid sequence of PKC1 shows high homology to PKCs of other filamentous fungi and all define a new subgroup of PKCs. All attempts to disrupt PKC1 failed, suggesting, but not proving, that disruption of PKC1 function is lethal. About 1 kb 3' of PKC1 is FAT1 encoding a putative bifunctional fatty acid transporter/very-long chain acyl-CoA synthetase. PMID- 9742700 TI - Molecular characterization of the erythromycin resistance plasmid pPV142 from Staphylococcus simulans. AB - The 2.5-kb erythromycin resistance (EmR) plasmid pPV142 of Staphylococcus simulans 13044 was isolated and characterized. Sequence analysis identified ORF1 and ORF2 encoding a 158-residue replication protein (Rep142) and a 244-residue erythromycin resistance protein (Erm, rRNA adenine N-6-methyltransferase), respectively. Structural analysis and Southern hybridization showed that the rep and ermM genes in pPV142 shared homology with the EmR plasmid pPV141 (2.4 kb) of S. chromogenes 3688 and other EmR plasmids known to exist in staphylococci and bacilli. Based on the presence of a 61-bp repeat upstream of the ermM gene, pPV142 is apparently a unique member of the pSN2 family of EmR plasmid able to express erythromycin resistance constitutively. PMID- 9742701 TI - Two novel plasmid-mediated cefotaxime-hydrolyzing beta-lactamases (CTX-M-5 and CTX-M-6) from Salmonella typhimurium. AB - Two novel plasmid-mediated beta-lactamases (CTX-M-5 and CTX-M-6) produced by Salmonella typhimurium clinical strains were characterized. The enzymes exhibited a pI of 8.4, hydrolyzed oxyimino-beta-lactams and were susceptible to mechanism based beta-lactamase inhibitors. The respective bla genes were cloned and sequenced. The deduced amino acid sequences showed a high degree of homology with those of the previously described plasmid class A CTX-M-type enzymes and appeared related to the chromosomal beta-lactamases of Klebsiella oxytoca. PMID- 9742702 TI - Distribution of the antiseptic-resistance gene qacE delta 1 in gram-positive bacteria. AB - The distribution of the antiseptic-resistance genes qacE and qacE delta 1, originally isolated from Gram-negative bacteria, was studied in a large number of Gram-positive bacteria by a method that included the polymerase chain reaction. A total of 151 strains of Staphylococcus and Enterococcus, isolated from clinical sources and obtained from the Japanese Collection of Microorganisms, was used in this analysis. We found the qacE delta 1 gene in 36 of 103 strains of Staphylococcus and in nine of 48 strains of Enterococcus. All of the strains in which we detected the qacE delta 1 gene were clinical isolates. The qacE gene was not detected in any of the strains examined in this study. The nucleotide sequences of the qacE delta 1 genes from the strains of Staphylococcus and Enterococcus were identical to that of the gene located on integron InC in Pseudomonas aeruginosa. These results indicate that the antiseptic-resistance gene qacE delta 1 is present in Gram-positive, as well as Gram-negative, bacteria. PMID- 9742703 TI - Methionine formation from alpha-ketomethiobutyrate in the trypanosomatid Crithidia fasciculata. AB - Methionine consumed during the synthesis of polyamines can be recycled in most organisms by a unique pathway wherein the final step is the transaminative conversion of alpha-ketomethiobutyrate to methionine (KMAT activity). In the trypanosomatid Crithidia fasciculata, three separate aminotransferases (KMAT-A, B, -T) were found to catalyse this activity. All three aminotransferases were found to utilise aromatic amino acids as the amino donor for the KMAT reaction, but KMAT-A functioned optimally with histidine and KMAT-B with arginine as amino donors. KMAT-T was found to operate best with aromatic amino acids and glutamate as amino donors, and was also found to catalyse aspartate aminotransferase and tyrosine aminotransferase activities. Amino acid sequencing of internal peptides from KMAT-T yielded a sequence with very high identity to vertebrate, cytosolic aspartate aminotransferase. As pig heart cytosolic aspartate and alanine aminotransferases were found to be unable to catalyse KMAT activity, the crithidial enzyme appears to be an aspartate aminotransferase with unusual catalytic properties. Inhibition studies on C. fasciculata homogenates showed that carboxymethoxylamine, canaline, and nitrophenylalanine were effective inhibitors of total KMAT activity (63-100% inhibition at 1 mM in the presence of 1 mM alpha-ketomethiobutyrate and 30 mM total amino acid as substrates) and the individual, isolated enzymes. At 1 mg ml-1, canaline was found to inhibit cell growth in vitro by 62%, and carboxymethoxylamine caused cell death within 24 h. PMID- 9742704 TI - Plasmid content and localization of the genes encoding the denitrification enzymes in two strains of Rhodobacter sphaeroides. AB - Plasmid content and localization of the genes encoding the reductases of the denitrification pathway were determined in the photosynthetic bacterium Rhodobacter sphaeroides forma sp. denitrificans by transverse alternating-field electrophoresis (TAFE) and hybridization with digoxigenin-labeled homologous probes. Two large plasmids of 102 and 115 kb were found. The genes encoding the various reductases are not clustered on a single genetic unit. The nap locus (localized with a napA probe), the nirK gene and the norCB genes encoding the nitrate, nitrite and nitric oxide reductases, respectively, were found on different AseI and SnaBI digested chromosomal DNA fragments, whereas the nos locus (localized with a nosZ probe), encoding the nitrous oxide reductase, was identified on the 115-kb plasmid. Furthermore, the genes encoding two proteins of unknown function, one periplasmic and the other cytoplasmic, but whose synthesis is highly induced by nitrate, were found on a different chromosomal fragment. For comparison, the same experiments were carried out on the well-characterized strain Rhodobacter sphaeroides 2.4.1. PMID- 9742705 TI - Cloning and expression of the Campylobacter jejuni lon gene detected by RNA arbitrarily primed PCR. AB - Fingerprinting of RNA by arbitrarily primed PCR was used to identify a heat inducible gene in Campylobacter jejuni. Comparing RNA fingerprints from C. jejuni cells before and after 20 min of heat shock at 48 degrees C, a differentially amplified PCR product was identified which displayed a high degree of homology to bacterial lon genes. By screening C. jejuni genomic libraries, the entire lon gene was cloned and sequenced. It encodes a protein of 791 amino acids with a calculated molecular mass of 90.2 kDa. Alignment of the Lon amino acid sequence with that of other bacterial species revealed an overall identity of up to 56.6% (Helicobacter pylori). Northern and RNA dot blot experiments confirmed heat induction of the C. jejuni lon gene, revealing a maximum 6-8-fold increase in the level of specific mRNA. PMID- 9742706 TI - Development of an integrative DNA transformation system for the yeast Candida utilis. AB - We report here the development of an auxotrophic transformation system for the food yeast Candida utilis. To facilitate molecular studies in Candida utilis, we isolated auxotrophic strains for uracil biosynthesis by the combination of NTG mutagenesis and 5-fluorotic acid (FOA) selection. The ura-mutation could be functionally complemented by the homologous URA3 gene. We used both, LiAc and electroporation methods to direct insertions at the ura3 locus through homologous recombination. PMID- 9742707 TI - In vitro genotypic variation of Campylobacter coli documented by pulsed-field gel electrophoretic DNA profiling: implications for epidemiological studies. AB - Six isolates of Campylobacter coli from different pig herds were subcultured up to 50 times over a 6-month period and DNA samples suitable for pulsed-field gel electrophoretic (PFGE) profiling prepared at regular (1, 20, 40 and 50 passages) intervals. In 5/6 strains, changes in the banding patterns of Sma1, Sal1 and/or BamH1 digests were observed. In one such strain the differences were considered artifactual. However, significant alterations in PFGE profiles between subcultures of four strains were seen, irrespective of the restriction enzyme used. Spontaneous intramolecular genomic rearrangements were considered the most likely mechanism for the changes observed. A numerical analysis based upon the combined distribution of Sma1- and sal1-derived fragments clustered most strain subcultures together, with the exception of those from one isolate which were divided into two clusters. The effect of spontaneous genetic change on PFGE profiles must be considered when evaluating strain relationships. Numerical techniques may aid data interpretation but results must be evaluated cautiously. PMID- 9742709 TI - Comparison of ARDRA and recA-RFLP analysis for genomic species identification of Acinetobacter spp. AB - The genus Acinetobacter is subdivided into genospecies on the basis of DNA relatedness of strains. Phenotypic tests are insufficient to identify the genospecies to which an isolate belongs. The effectiveness of two previously described PCR-based methods for genospeciating Acinetobacter spp. was compared using a group of 32 well-characterised strains representing six genospecies. Amplified ribosomal DNA restriction analysis (ARDRA) correctly identified all 32 strains. Using restriction fragment length polymorphism (RFLP) of recA PCR amplimers, only six of the 32 strains were correctly identified. Heterogeneity in the recA gene sequence was demonstrated within five of the genospecies. ARDRA proved to be a reliable method whereas analysis of recA RFLP profiles did not enable the genospecies of most of the isolates of Acinetobacter spp. to be determined. PMID- 9742710 TI - Fine structure of hepatitis B virus surface antigen produced by recombinant yeast: comparison with HBsAg of human origin. AB - The ultrastructure of hepatitis B virus surface antigen (HBsAg) particles produced by recombinant yeast cells was examined using high-resolution negative staining, and ice embedding, electron microscopy. With negative staining, the HBsAg particles were spherical to slightly ovoid with a mean diameter of 27.5 nm and consisted of many subunits each 4 nm in diameter. Subunits were marked with a minute central pore. With ice embedding, particles were mostly spherical to ovoid, with a mean diameter of 23.7 nm and a 7-8 nm thick cortex surrounding an electron translucent core. Human HBsAg particles, examined using the same methods, were smaller, apparently because of molecular differences in polypeptide structure. PMID- 9742711 TI - Co-detection of Helicobacter pylori and of its resistance to clarithromycin by PCR. AB - Our aim was to develop a rapid molecular test based on polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) and making it possible to detect Helicobacter pylori directly from gastric biopsy samples, and to test its susceptibility to clarithromycin. A 629-bp fragment of the 23S rRNA gene of H. pylori was amplified by PCR and the mutations responsible for clarithromycin resistance were detected with Bsa1 and Bbs1 restriction endonucleases. Thirty five gastric samples were tested in parallel by standard microbiologic methods (culture and clarithromycin susceptibility testing with E-test strips) and by PCR RFLP. The 10 culture-negative samples were also PCR-negative. Sixteen out of the 25 culture-positive samples (64%) were PCR-positive. RFLP analysis could be done in 12 cases and the results were in agreement with those of the E-test: susceptibility in five cases, resistance in seven (six A2144G mutations and one A2143G mutation). PMID- 9742713 TI - 4-(5,6-dimethoxy-2-phthalimidinyl)phenylsulfonyl semipiperazide as a fluorescent labelling reagent for determination of carboxylic acids in precolumn high performance liquid chromatography. AB - A fluorescent labelling reagent, 4-(5,6-dimethoxy-2-phthalimidinyl)phenylsulfonyl semipiperazide, was designed for the determination of carboxylic acids by precolumn HPLC. The reagent was reacted with carboxylic acids at 100 degrees C for 15 min in the presence of an activating reagent (a mixture of triphenylphosphine, 2,2'-dipyridyldisulfide and pyridine) and produced highly fluorescent derivatives, which were separated on a reversed-phase column by fluorescence measurement at 317 nm (excitation) and 380 nm (emission). The detection limits were 4-12 fmol/injection. The reagent was used for HPLC assays of long chain fatty acids in human serum by isocratic elution. PMID- 9742712 TI - A proposal to transfer Vibrio marinus (Russell 1891) to a new genus Moritella gen. nov. as Moritella marina comb. nov. AB - The taxonomic positions of Vibrio marinus and 11 related natural isolates were examined. Their phylogenetic positions on the basis of almost complete 16S rRNA sequences were determined by maximum likelihood, maximum parsimony and neighbor joining analyses. V. marinus and the 11 isolates fell into a single cluster that was clearly distinct from other genera. There is now strong evidence that V. marinus should be reclassified as Moritella marina gen. nov., comb. nov. PMID- 9742714 TI - Comparison of supercritical fluid extraction and Soxhlet extraction for the determination of polychlorinated biphenyls in environmental matrix standard reference materials. AB - Supercritical fluid extraction (SFE) was compared to traditional Soxhlet extraction for the determination of polychlorinated biphenyl congeners in three standard reference materials: SRM 1941a (Organics in Marine Sediment), SRM 1944 (New York/New Jersey Waterway Sediment) and SRM 2974 [Organics in Mussel Tissue (Mytilus edulis) (Freeze-Dried)]. The concentrations determined using SFE compared well with the certified concentrations for the majority of the polychlorinated biphenyl congeners. PMID- 9742715 TI - Memory monitoring by animals and humans. AB - The authors asked whether animals and humans would use similarly an uncertain response to escape indeterminate memories. Monkeys and humans performed serial probe recognition tasks that produced differential memory difficulty across serial positions (e.g., primacy and recency effects). Participants were given an escape option that let them avoid any trials they wished and receive a hint to the trial's answer. Across species, across tasks, and even across conspecifics with sharper or duller memories, monkeys and humans used the escape option selectively when more indeterminate memory traces were probed. Their pattern of escaping always mirrored the pattern of their primary memory performance across serial positions. Signal-detection analyses confirm the similarity of the animals' and humans' performances. Optimality analyses assess their efficiency. Several aspects of monkeys' performance suggest the cognitive sophistication of their decisions to escape. PMID- 9742716 TI - Aging and source monitoring: cognitive processes and neuropsychological correlates. AB - This study shows that relative to younger adults, older adults are more adversely influenced by similar items when judging a memory's source, and the phenomenal features of their correctly and incorrectly attributed memories have greater overlap. The authors argue in accordance with the source monitoring framework that this age-related impairment in source accuracy is related to processes involved in binding features into complex memories and those involved in accessing and evaluating contextual features of memories. These processes are linked to medial temporal and frontal brain regions, respectively, as evidenced by correlations in older adults between source accuracy and neuropsychological tests often used to assess medial temporal and frontal function. The results suggest that adequate feature binding is particularly important when items from different source share similar features and access-evaluation processes are particularly important after a delay. PMID- 9742717 TI - Replicable unconscious semantic priming. AB - In 4 experiments, subjects classified visually presented target words as pleasant unpleasant words or male-female first names. Prime words were similar (congruent) or dissimilar (incongruent) in meaning to targets. Brief duration of prime words (17, 33, or 50 ms), along with pre- and postmasking, prevented most subjects from perceiving their physical and semantic properties. By constraining response latencies to fall within a response window--a narrow time band that occurred earlier than subjects would ordinarily respond--these experiments consistently produced subliminal priming effects, indicated by greater error rates for incongruent than congruent priming trials. This conclusion was confirmed by analyzing magnitude of priming as a regression function of prime perceptibility using the method of A. G. Greenwald, M. R. Klinger, and E. S. Schuh (1995). The data of each experiment passed their significant-intercept criterion for demonstrating unconscious cognition. PMID- 9742718 TI - On demonstrating unconscious perception: comment on Draine and Greenwald (1998) AB - S. C. Draine and A. G. Greenwald (1998) have described a methodology based on regression analysis for demonstrating unconscious perception. They have suggested that their methodology represents a major improvement over existing methodologies. An analysis of their methodology reveals that it is closely related to the classic dissociation paradigm. As such, interpretation of their results is compromised by the same issues concerning the measurement of awareness that have plagued all previous attempts to use the dissociation paradigm to demonstrate unconscious perception in the complete absence of conscious perception. PMID- 9742719 TI - Correcting for measurement error in detecting unconscious cognition: comment on Draine and Greenwald (1998) AB - A. G. Greenwald, M. R. Klinger, and E. S. Schuh (1995) have proposed a regression method for detecting unconscious cognition in experiments that obtain measures of indirect and direct effects of stimuli with suspected unconscious effects. Their indirect-on-direct-measure regression approach can produce misleading evidence for indirect effects in the absence of direct effects when the direct-effect measure has typical measurement error. This article describes an errors-in variables variant of the regression method that corrects for error in the direct effect measure. Applied to the uses of the regression method by S. C. Draine and A. G. Greenwald (1998) in this issue, the errors-in-variables method affirms substantial evidence for indirect effects in the absence of direct effects. PMID- 9742720 TI - Distinguishing unconscious from conscious cognition--reasonable assumptions and replicable findings: reply to Merikle and Reingold (1998) and Dosher (1998) AB - S. C. Draine and A. G. Greenwald (1998) demonstrated replicable unconscious semantic priming by combining a response window procedure, which increases priming effects by requiring rapid responding, and a regression analysis in which the regression intercept is a marker for unconscious cognition. The commentaries by B. A. Dosher (1998) and by P. M. Merikle and E. M. Reingold (1998) raise two questions about conclusions based on these methods: (a) Did Draine and Greenwald (1998) demonstrate an indirect effect (subliminal priming) in the absence of a direct effect (i.e., visibility of the subliminal priming words)? and (b) Did Draine and Greenwald (1998) demonstrate dissociation of conscious from unconscious cognition? The first question has reassuring responses that are reviewed here. The second question is answered by pointing out that although Draine and Greenwald (1998) did not claim to have established such dissociation, they provided data that advance the plausibility of that conclusion. PMID- 9742721 TI - The diffuse interstellar bands: a tracer for organics in the diffuse interstellar medium? AB - The diffuse interstellar bands (DIBs) are absorption bands seen in the spectra of stars obscured by interstellar dust. DIBs are recognized as a tracer for free, organic molecules in the diffuse interstellar medium (ISM). The potential molecular carriers for the DIBs are discussed with an emphasis on neutral and ionized polycyclic aromatic hydrocarbons (PAHs) for which the most focused effort has been made to date. From the combined astronomical, laboratory and theoretical study, it is concluded that a distribution of free neutral and ionized complex organics (PAHs, fullerenes, unsaturated hydrocarbons) represents the most promising class of candidates to account for the DIBs. The case for aromatic hydrocarbons appears particularly strong. The implied widespread distribution of complex organics in the diffuse ISM bears profound implications for our understanding of the chemical complexity of the ISM, the evolution of prebiotic molecules and its impact on the origin and the evolution of life on early Earth through the exogenous delivery (cometary encounters and metoritic bombardments) of prebiotic organics. PMID- 9742722 TI - Extraterrestrial organic matter: a review. AB - We review the nature of the widespread organic material present in the Milky Way Galaxy and in the Solar System. Attention is given to the links between these environments and between primitive Solar System objects and the early Earth, indicating the preservation of organic material as an interstellar cloud collapsed to form the Solar System and as the Earth accreted such material from asteroids, comets and interplanetary dust particles. In the interstellar medium of the Milky Way Galaxy more than 100 molecular species, the bulk of them organic, have been securely identified, primarily through spectroscopy at the highest radio frequencies. There is considerable evidence for significantly heavier organic molecules, particularly polycyclic aromatics, although precise identification of individual species has not yet been obtained. The so-called diffuse interstellar bands are probably important in this context. The low temperature kinetics in interstellar clouds leads to very large isotopic fractionation, particularly for hydrogen, and this signature is present in organic components preserved in carbonaceous chondritic meteorites. Outer belt asteroids are the probable parent bodies of the carbonaceous chondrites, which may contain as much as 5% organic material, including a rich variety of amino acids, purines, pyrimidines, and other species of potential prebiotic interest. Richer in volatiles and hence less thermally processed are the comets, whose organic matter is abundant and poorly characterized. Cometary volatiles, observed after sublimation into the coma, include many species also present in the interstellar medium. There is evidence that most of the Earth's volatiles may have been supplied by a 'late' bombardment of comets and carbonaceous meteorites, scattered into the inner Solar System following the formation of the giant planets. How much in the way of intact organic molecules of potential prebiotic interest survived delivery to the Earth has become an increasingly debated topic over the last several years. The principal source for such intact organics was probably accretion of interplanetary dust particles of cometary origin. PMID- 9742723 TI - A search for extraterrestrial amino acids in carbonaceous Antarctic micrometeorites. AB - Antarctic micrometeorites (AMMs) in the 100-400 microns size range are the dominant mass fraction of extraterrestrial material accreted by the Earth today. A high performance liquid chromatography (HPLC) based technique exploited at the limits of sensitivity has been used to search for the extraterrestrial amino acids alpha-aminoisobutyric acid (AIB) and isovaline in AMMs. Five samples, each containing about 30 to 35 grains, were analyzed. All the samples possess a terrestrial amino acid component, indicated by the excess of the L-enantiomers of common protein amino acids. In only one sample (A91) was AIB found to be present at a level significantly above the background blanks. The concentration of AIB (approximately 280 ppm), and the AIB/isovaline ratio (> or = 10), in this sample are both much higher than in CM chondrites. The apparently large variation in the AIB concentrations of the samples suggests that AIB may be concentrated in rare subset of micrometeorites. Because the AIB/isovaline ratio in sample A91 is much larger than in CM chondrites, the synthesis of amino acids in the micrometeorite parent bodies might have involved a different process requiring an HCN-rich environment, such as that found in comets. If the present day characteristics of the meteorite and micrometeorite fluxes can be extrapolated back in time, then the flux of large carbonaceous micrometeorites could have contributed to the inventory of prebiotic molecules on the early Earth. PMID- 9742724 TI - Observation of indigenous polycyclic aromatic hydrocarbons in 'giant' carbonaceous antarctic micrometeorites. AB - Two-step laser desorption/laser ionization mass spectrometry (microL2 MS) was used to establish the nature and mass distribution of polycyclic aromatic hydrocarbons (PAHs) in fragments of fifteen 'giant' (approximately 200 microns) carbonaceous Antarctic micrometeorites (AMMs). Detectable concentrations of PAHs were observed in all AMMs showing a fine-grained matrix. The range of integrated PAH signal intensities varied between samples by over two orders of magnitude. No evidence of contamination whilst in the Antarctic environment could be found. The dramatic variation of both PAH signal intensities and mass distributions between AMMs along with comprehensive contamination checks demonstrates that particles are not exposed to terrestrial PAHs at or above detection limits, either subsequent, during or prior to collection. Comparison of the observed PAH distributions with those measured in three carbonaceous chondrites [Orgueil (CI1), Murchison (CM2) and Allende (CV3)] under identical conditions demonstrated that marked differences exist in the trace organic composition of these two sources of extraterrestrial matter. In general, AMMs show a far richer distribution of unalkylated 'parent' PAHs with more extended alkylation series (replacement of -H with -(CH2)n-H; n = 1, 2, 3 ...). The degree of alkylation loosely correlates with a metamorphic index that represents the extent of frictional heating incurred during atmospheric entry. A search for possible effects of the chemical composition of the fine-grain matrix of host particles on the observed PAH distributions reveals that high degrees of alkylation are associated with high Na/Si ratios. These results, in addition to other observations by Maurette, indicate that 'giant' micrometeorites survive hypervelocity (> or = 11 km s-1) atmospheric entry unexpectedly well. Because such micrometeorites are believed to represent the dominant mass fraction of extraterrestrial material accreted by the Earth, they may have played a significant role in the prebiotic chemical evolution of the early Earth through the delivery of complex organic matter to the surface of the planet. PMID- 9742725 TI - Biogenic catalysis of soil formation on Mars? AB - The high iron abundance and the weak ferric iron spectral features of martian surface material are consistent with nanophase (nm-sized) iron oxide minerals as a major source of iron in the bright region soil on Mars. Nanophase iron oxide minerals, such as ferrihydrite and schwertmannite, and nanophase forms of hematite and goethite are formed by both biotic and abiotic processes on Earth. The presence of these minerals on Mars does not indicate biological activity on Mars, but it does raise the possibility. This work includes speculation regarding the possibility of biogenic soils on Mars based on previous observations and analyses. A remote sensing goal of upcoming missions should be to determine if nanophase iron oxide minerals, clay silicates and carbonates are present in the martian surface material. These minerals are important indicators for exobiology and their presence on Mars would invoke a need for further investigation and sample return from these sites. PMID- 9742726 TI - Formation of RNA oligomers on montmorillonite: site of catalysis. AB - Certain montmorillonites catalyze the self condensation of the 5' phosphorimidazolide of nucleosides in pH 8 aqueous electrolyte solutions at ambient temperatures leading to formation of RNA oligomers. In order to establish the nature of the sites on montmorillonite responsible for this catalytic activity, oligomerization reactions were run with montmorillonites which had been selectively modified (I) at the edges by (a) fluoride treatment, (b) silylation, (c) metaphosphate treatment of the anion exchange sites (II) in the interlayer by (a) saturation with quaternary alkylammonium ions of increasing size, (b) aluminum polyoxo cations. High pressure liquid chromatography, HPLC, analysis of condensation products for their chain lengths and yields indicated that modification at the edges did not affect the catalytic activity to a significant extent, while blocking the interlayer strongly inhibited product formation. PMID- 9742727 TI - A fission-fusion origin for life. AB - To develop a comprehensive 'cells-first' approach to the origin of life, we propose that protocells form spontaneously and that the fission and fusion of these protocells drives the dynamics of their evolution. The fitness criterion for this evolution is taken to be the the stability (conservation) of domains in the protocellular membrane as determined by non-covalent molecular associations between the amphiphiles of the membrane and a subset of the macromolecules in the protocell. In the presence of a source of free energy the macromolecular content of the protocell (co-)evolves as the result of (domain-dependent) membrane catalysed polymerisation of the prebiotic constituents delivered to the protocell by fusion. The metabolism of the cell therefore (co-)evolves on a rugged fitness landscape. We indicate how domain evolution with the same fitness criterion can potentially give rise to coding. Membrane domains may therefore provide the link between protocells and the RNA/DNA-world. PMID- 9742728 TI - The role of gene duplication in the evolution of purine nucleotide salvage pathways. AB - Purine nucleotides are formed de novo by a widespread biochemical route that may be of monophyletic origin, or are synthesized from preformed purine bases and nucleosides through different salvage pathways. Three monophyletic sets of purine salvage enzymes, each of which catalyzes mechanistically similar reactions, can be identified: (a) adenine-, xanthine-, hypoxanthine- and guanine phosphoribosyltransferases, which are all homologous among themselves, as well as to nucleoside phosphorylases; (b) adenine deaminase, adenosine deaminase, and adenosine monophophate deaminase; and (c) guanine reductase and inosine monophosphate dehydrogenase. These homologies support the idea that substrate specificity is the outcome of gene duplication, and that the purine nucleotide salvage pathways were assembled by a patchwork process that probably took place before the divergence of the three cell domains (Bacteria, Archaea, and Eucarya). Based on the ability of adenine PRTase to catalyze the condensation of PRPP with 4-aminoimidazole-5-carboxamide (AICA), a simpler scheme of purine nucleotide biosynthesis is presented. This hypothetical route requires the prior evolution of PRPP biosynthesis. Since it has been argued that PRPP, nucleosides, and nucleotides are susceptible to hydrolysis, they are very unlikely prebiotic compounds. If this is the case, it implies that many purine salvage pathways appeared only after the evolution of phosphorylated sugar biosynthetic pathways made ribosides available. PMID- 9742731 TI - About various definitions of life. AB - The old question of a definition of minimal life is taken up again at the aim of providing a forum for an updated discussion. Briefly discussed are the reasons why such an attempt has previously encountered scepticism, and why such an attempt should be renewed at this stage of the inquiry on the origin of life. Then some of the definitions of life presently used are cited and briefly discussed, starting with the definition adopted by NASA as a general working definition. It is shown that this is too limited if one wishes to provide a broad encompassing definition, and some extensions of it are presented and discussed. Finally it is shown how the different definitions of life reflect the main schools of thought that presently dominate the field on the origin of life. PMID- 9742729 TI - Evolution of the structure and chromosomal distribution of histidine biosynthetic genes. AB - A database of more than 100 histidine biosynthetic genes from different organisms belonging to the three primary domains has been analyzed, including those found in the now completely sequenced genomes of Haemophilus influenzae, Mycoplasma genitalium, Synechocystis sp., Methanococcus jannaschii, and Saccharomyces cerevisiae. The ubiquity of his genes suggests that it is a highly conserved pathway that was probably already present in the last common ancestor of all extant life. The chromosomal distribution of the his genes shows that the enterobacterial histidine operon structure is not the only possible organization, and that there is a diversity of gene arrays for the his pathway. Analysis of the available sequences shows that gene fusions (like those involved in the origin of the Escherichia coli and Salmonella typhimurium hisIE and hisB gene structures) are not universal. In contrast, the elongation event that led to the extant hisA gene from two homologous ancestral modules, as well as the subsequent paralogous duplication that originated hisF, appear to be irreversible and are conserved in all known organisms. The available evidence supports the hypothesis that histidine biosynthesis was assembled by a gene recruitment process. PMID- 9742730 TI - A search for extraterrestrial eukaryotes: physical and paleontological aspects. AB - Physical and biochemical aspects of a proposed search for extraterrestrial eukaryotes (SETE) are considered. Such a program should approach the distinction between a primitive eukaryote and an archaebacteria. The emphasis on gene silencing suggests a possible assay suitable for a robotic investigation of eukaryoticity, so as to be able to decide whether the first steps towards eukaryogenesis have been taken in an extraterrestrial planet, or satellite. The experiment would consist of searching for cellular division and the systematic related delay in replication of heterochromatic chromosome segments. It should be noticed that the direct search for a membrane-bounded set of chromosomes does not necessarily determine eukaryotic identity, as there are prokaryotes that have membrane-bounded nucleoids. A closer look at the protein fraction of chromatin (mainly histones) does not help either, as there are some eukaryotes that may lack histones; there are also some bacteria as well as archaebacteria with histone-like proteins in their nucleoids. Comments on the recent suggestion of possible environments for a SETE program are discussed: the deep crust of Mars, and the Jovian satellite Europa, provided the existence of an ocean under its ice covered surface is confirmed by the current Galileo mission. PMID- 9742732 TI - [Reproducible EEG alpha-rhythm patterns in solving psychological tasks]. PMID- 9742734 TI - [Formation of concrete visual thinking in infants with visual defects in the first year of life]. PMID- 9742733 TI - [Origin of inverse spatial distribution of alpha rhythm frequencies]. PMID- 9742736 TI - [Central mechanisms of regulation of voluntary movements in 6-10-year-old children. II. Electrophysiologic analysis of movement performance in right-handed children]. PMID- 9742735 TI - [The psychophysiologic characteristics of individual temperamental features of children with early visual deprivation in the first year of life]. PMID- 9742737 TI - [Is there a locomotor generator in man?]. PMID- 9742738 TI - [Gradation of hypoxic states in man during intense muscular activity]. PMID- 9742739 TI - [Factor analysis of manual preference in various actions in children 7-10 years old]. PMID- 9742741 TI - [Arterial blood flow in leg vessels in healthy men of various age groups]. PMID- 9742740 TI - [Clinical-physiologic characteristics of carpal tunnel neuropathies]. PMID- 9742742 TI - [The diagnostic effectiveness of methods for quantitative assessment of individual health]. PMID- 9742743 TI - [Morpho-functional evaluation of health status in adolescents]. PMID- 9742744 TI - [Physiological reactions of the female respiratory system to additional breathing resistance]. PMID- 9742745 TI - [Nutritional risk factors in the development of cardiovascular diseases under the natural climatic conditions of Turkmenistan]. PMID- 9742746 TI - [Parameters of the cardiorespiratory system in athletes of various ages]. PMID- 9742747 TI - [Development in human ontogenesis of the mechanisms of microcirculatory adaptation to dynamic local loads]. PMID- 9742748 TI - [Inhibition of platelet function by biogenic chloramines]. PMID- 9742749 TI - [Content of lipid peroxidation products in the urine of patients with ischemic heart disease]. PMID- 9742750 TI - [Capnogram and cyclic organization of sleep in full-term and premature newborns]. PMID- 9742751 TI - [Reserve capacities of the kallikrein-kinin system of the blood in the normal course of pregnancy]. PMID- 9742752 TI - [Evaluation of erythrocyte beta-adrenoreactivity in parturient women]. PMID- 9742754 TI - [Iron metabolic indices im man studied under conditions of head-down tilt hypokinesia]. PMID- 9742755 TI - [The Chediak-Higashi syndrome]. AB - The Chediak-Higashi syndrome is described. The morphology of the blood and bone marrow in a girl aged 4.5 months [correction of years] is described in detail. Hematological, immunological, and karyological characteristics of her parents are presented. PMID- 9742756 TI - [General recommendations for laboratory work in studying blood coagulability]. PMID- 9742757 TI - [The interrelationship of the intensity of free-radical oxidation to the level of serum bilirubin in lesions of the hepatobiliary system]. AB - The level of lipid peroxidation (LPO) products increases in the blood of patients with chronic diseases of the liver and acute cholecystitis with mechanical jaundice. The values normalize after surgery for acute cholecystitis, whereas therapy for chronic diseases of the liver virtually does not correct them. These data demonstrate the relationship between intensity of free-radical oxidation and severity of hepatobiliary process and between plasma content of LPO products and level of serum bilirubin. PMID- 9742758 TI - [The use of noninvasive biological means in assessing lipids in children]. AB - Gas chromatographic analysis of the lipid fatty acid spectrum of blood (serum and plasma), sweat, and exhaled air condensate in children with acute pneumonia and dermatitis caused by alimentary allergies showed good correlation of the results. Therefore, biological objects obtained by noninvasive methods can be used for studies of lipid metabolism in children. PMID- 9742759 TI - [The use of electrophoretic methods for determining modified proteins in diabetic patients]. AB - Modified proteins were determined by isoelectric focusing in borate-polyol system with subsequent colorimetry (micromethod) and electrophoresis of blood serum on paper with subsequent TCA-ethanol treatment. Increased levels of glycated hemoglobin and modified albumin and changed light absorbance of glycated albumin were detected. The levels of glycated hemoglobin assessed by the micromethod and colorimetry without calibration did not correlate. PMID- 9742760 TI - [An improvement in the methods for the microbiological study of the blood from patients with suppurative-septic infections]. AB - The new method for microbiological analysis of blood in patients with pyoseptic infections is based on separate inoculations of plasma, erythrocyte mass, and leukocyte layer in nutrient medium providing conditions for growth of a wide spectrum of aerobes and obligate anaerobes. The method in general ensures a more rapid result and permits differentiation between true and false-positive bacteremia and between stages of septic process (bacteremia and septicemia). PMID- 9742761 TI - [A replaceable nonfrictional membrane filter for the prevention of cell culture contamination]. AB - A changeable membrane filter preventing the friction between moving parts and the membrane during tight screwing of a culture flask is proposed. PMID- 9742762 TI - [The laboratory diagnosis of spring-summer infections in Yekaterinburg in the epidemic season for tick-borne encephalitis]. AB - The authors propose a comprehensive approach to laboratory diagnosis of seasonal transmissible infections, based on modern methods permitting etiological deciphering of disease. A universal diagnostic algorithm notably accelerated the laboratory diagnosis due to cutting the period between collection of material from a patient and consecutive screening for antibodies to agents of tick-borne encephalitis, Lyme disease, and California encephalitis. PMID- 9742763 TI - [The use of a species-specific erythrocytic diagnostic agent for detecting pseudotuberculosis and yersiniosis]. PMID- 9742764 TI - [The myelodysplastic syndrome (a lecture)]. PMID- 9742765 TI - [The determination of the volume (weight) of an inoculate applied using the Semenikhina-designed replicator]. AB - The weight (volume) of inoculate applied with Semenikhina's replicator stamp with 1, 2, and 3-mm needles is determined. The method is sufficiently precise for microbiological screenings. The volume of inoculate depends on the diameter and geometrical shape of the needle and characteristics of inoculated substance and surface onto which it is applied. PMID- 9742766 TI - [Ways to decrease inhibition of the polymerase chain reaction by the components of the clinical specimens]. AB - Inhibition of the polymerase chain reaction (PCR) by components of the analyzed specimen is an important problem in analysis of clinical material by this method. The inhibitory activity of clinical material can be decreased by additional purification of DNA by gel filtration on microcolumns. The method was developed using blood-containing specimens and tried on many clinical specimens. Commercial PCR kit for detecting Mycobacterium tuberculosis was used. The proposed method for DNA purification notably improves the efficacy of detecting the agents by PCR. PMID- 9742767 TI - [Laboratory service: problems and their solution]. PMID- 9742768 TI - [Standards bases for the activities of clinical diagnostic laboratories today]. PMID- 9742769 TI - [Current concepts on the etiology and pathogenesis of systemic vasculitis: the role of infection and genetic predisposition (I)]. PMID- 9742770 TI - [Losartan--a nonpeptide antagonist of angiotensin-II receptors in the treatment of heart failure]. PMID- 9742771 TI - [Rheumatoid arthritis and osteoporosis]. PMID- 9742772 TI - [Ventricular late potentials and ventricular arrhythmias in noncoronarogenic myocardial diseases]. AB - Hemodynamic parameters and survival were studied in 48 patients with noncoronarogenic myocardial diseases (NCMD). The examination included high resolution electrocardiography and Holter monitoring of cardiac rhythm. Deterioration of pump capacity of the heart was associated with high-grade arrhythmias (4B and higher) while ventricular late potentials were more characteristic for patients with severe hemodynamic disorders and, in particular, with dilated cardiomyopathy. The prognosis in NCMD is based on clinical evidence on functional class of the heart failure, parameters of central and intrapulmonary hemodynamics. PMID- 9742773 TI - [The use of the antioxidant coenzyme Q10 as a cytoprotection variant in ischemic heart disease]. PMID- 9742774 TI - [The characteristics of the clinical manifestations and treatment of chronic obstructive bronchitis in patients with hepatobiliary system diseases and pseudoallergy]. AB - Routine clinical, laboratory, instrumental and immunological examinations in 105 patients have established that hepatobiliary disorders occur 2.2 times more frequently in patients with chronic obstructive bronchitis (COB) and pseudoallergy than in those without pseudoallergy. To correct negative effects of pseudoallergy, conventional therapy was combined with enterosorbent polyphepan, spasmolytic nospa and cholagogic allochol used alone or in different combinations. The highest benefit for COB patients was achieved in using combination of polyphepan + nospa + allochol. PMID- 9742775 TI - [The rheological properties of the blood in patients with tick-borne typhus]. AB - Rheological characteristics of blood were studied in 50 patients with north Asian tick typhoid varying in severity and duration. In addition to standard diagnosis- verification tests, blood macrorheological and microrheological examinations were made (blood viscosity, hematocrit and red cell deformability and aggregation, respectively). It was found that both in the acute disease and recovery there were changes in blood and plasma viscosity red cell hardness. This suggests introduction of rheoprotectors and desaggregants into combined therapy of the above typhoid. PMID- 9742776 TI - [The conservative therapy of acute esophagogastric hemorrhages in portal hypertension patients]. PMID- 9742777 TI - [The therapeutic potentials of Berotec-100 in bronchial asthma patients]. PMID- 9742779 TI - [The characteristics of the onset and course of bronchopulmonary pathology in persons who abuse alcohol]. PMID- 9742778 TI - [Sulodexide in the treatment of diabetic nephropathy]. AB - Sulodexide (Vessel 2F, Italy) was given to 20 patients with diabetes mellitus type I with initial nephropathy. Urine excretion of albumin significantly fell in 18 patients and remained so in 15 patients 6 weeks after the drug discontinuation. Plasma total cholesterol tended to lowering. No changes were registered in hemostasis and carbohydrate metabolism. The authors propose combined administration of sulodexide and inhibitors of angiotensin-converting enzyme for treatment and prevention of diabetic nephropathy progression. PMID- 9742780 TI - [Stereotypes in prescribing antiasthmatic treatment]. PMID- 9742782 TI - [Cardiac arrhythmias]. PMID- 9742781 TI - [An apathetic form of thyrotoxicosis]. PMID- 9742783 TI - [The hepatotoxicity of flutamide]. PMID- 9742784 TI - [The Williams-Campbell syndrome diagnosed in an adult female patient]. PMID- 9742785 TI - [A rare case of pronounced disorders of atrioventricular conduction combined with a permanent form of atrial flutter]. PMID- 9742786 TI - [The art of the clinician lecturer. 3. The planning of a lecture course]. PMID- 9742787 TI - [G. A. Zakhar'in: at the crossroads of clinical medicine (1)]. PMID- 9742788 TI - [N. N. Kirikov--the first professor and therapist of Saratov]. PMID- 9742790 TI - [Late outcome of operated acute aortic dissection: risk factors analyzed from autopsy findings]. AB - Based on analysis of 35 autopsy cases of aortic dissection, risk factors related to late outcomes, especially to aortic rupture after operation, are discussed. Pathological findings related to the entry into a persistent distal false lumen (PDFL) after surgical repair of aortic dissection are: 1) unresected initial intimal tear; 2) a new intimal tear, mainly related to the surgical suture; 3) disrupted aortic branches, and 4) penetrating atherosclerotic aortic ulcers (PAU). These also are common pathological findings in ulcer-like projections and may cause re-dissection. After operation, the pseudoneointima develops and covers the PDFL during 1 to 2 months, becoming stable thereafter. In the remaining thrombosed-type of dissection after operation, the hematomas become completely organized only when they are small. Late rupture of an aneurysm in both thrombosed and non-thrombosed types of dissection may be caused by new dissection or by re-dissection from the sites of entry listed above. The PAU is an especially important factor related to the aging of patients after the operation. The findings of adventitial accumulations of macrophages, which are known to express matrix metalloproteinases, at the sites of rupture suggested that these enzymes also contribute to the rupture of chronic aortic dissection. PMID- 9742791 TI - [Late results of acute aortic dissection: analysis of the patients longer than five years after the operation]. AB - Between 1985 and 1992, 28 patients of acute type A aortic dissection were operated on at our department. Our surgical strategy for this disease is "limited aortic resection", that is to avoid replacement of the entire arch except for the patients with arch tear that cannot be resected without total arch replacement. There were one operative mortality due to post transfusion GVHD, and ten late mortality (rupture of the residual dissecting aneurysm 3; complication of the late reoperation 3; cerebrovascular disease 2; pulmonary infection 2). Actuarial survival rate of all cases is 92.9%, 62.9%, and 58.4% at 1, 5, and 10 years, respectively. Comparing the patients whose primary tear was resected or not resected, there was no difference in the rate of residual dissection (12/16, 75% vs 5/6, 83.3%; primary tear resected vs not resected), the rate of late reoperation (3/16, 18.8% vs 1/6, 16.7%), nor actuarial survival rate (90.5% vs 100%, 66.7% vs %, 53.6% vs 71.4%, at 1, 5, 10 years, respectively). There were three cases with Marfan's syndrome, and all three cases died of the rupture of the residual dissection. We will follow the policy of the "limited aortic resection" unless the operative mortality of the entire arch replacement is proved as good as that of the ascending or hemiarch replacement. Because of the poor late results of the patients with Marfan's syndrome, entire arch replacement at the initial surgery and aggressive reoperation for the residual dissection is necessary. PMID- 9742792 TI - [Operative results and long-term prognosis of type A acute aortic dissection]. AB - Between 1983 and 1997, we operated upon 91 patients with type A acute aortic dissection. The dissection was localized in 22 patients and extensive in 69 patients. All patients underwent graft replacement and 61 (67%) patients underwent simultaneous replacement of ascending aorta and total aortic arch. The hospital mortality rate were 9% for the localized dissection and 21% for the extensive dissection. The actuarial survival rates in patients with localized dissection at 5 and at 10 years was 91% and 76%, whereas those in patients with extensive dissection at 5 and at 10 years was 68% and 62%. The freedom from dissection related death or reoperation in operative survivors with localized dissection at 5 and at 10 years was 100% and 83%, whereas those in patients with extensive dissection at 5 and at 10 years was 78% and 56%. The simultaneous replacement of ascending aorta and total aortic arch in patients with extensive dissection was effective to obliteration of the distal false channel, although this extended procedure has to be carefully adopted in high risk patients with associated complications such as acute dissection organ ischemia. PMID- 9742793 TI - [Late results of extended Stanford type A acute aortic dissection]. AB - From 1978 to July 1997, 140 patients with extended Stanford type A aortic dissection underwent surgical treatment. There were 77 acute and 63 chronic aortic dissections. The follow-up period of the 61 surviving patients in acute aortic dissection ranged from 2 to 164 months (mean, 56 months). During follow-up periods, 6 patients died of no dissection-related causes. The actuarial survival ratios in 77 patients were 77.2% at 3 years, 72.2% at 5 years and 62.6% at 10 years, respectively. There was no significant difference between acute aortic dissection and chronic aortic dissection in actuarial survival ratio. The ratio of the clotted false lumen in the distal aorta was lower and subsequent surgery was more frequent in patients with Marfan syndrome than those with non-Marfan syndrome. In extended type A aortic dissection, the observation of the residual false lumen using body CT scan was very important after surgery, especially in Marfan syndrome. If the false lumen is dilated, a subsequent surgery is needed to get good late results. PMID- 9742795 TI - [Early and late surgical mortality of acute type A aortic dissections by open distal anastomosis with deep hypothermic retrograde cerebral perfusion]. AB - We have analyzed the operative results and the long-term prognosis of Stanford type A acute aortic dissections. Between 1981 and 1997, 57 patients underwent surgical repairs of acute type A aortic dissection. In the earlier period (1981 1990; n = 21), almost all the operations were performed under aortic cross clamping with conventional cardiopulmonary bypass, while in the later period (1991-1997; n = 36), radical resections and graft replacements under open distal anastomosis with deep hypothermic retrograde cerebral perfusion (RCP) were performed in 24 patients (66.7%) and with selective cerebral perfusion in 3 (8.3%). The hospital mortality rates were 33.3% in the earlier period and 16.7% in the later period. Two (8.3%) of 24 patients employed RCP failed by preoperative rupture to pulmonary artery and myocardial infarction. Reoperations for enlargement of the remained false lumen were performed in 4 in the earlier period, of whom 2 patients were dead. In the later period, 7 patients were reoperated on, and all patients survived. Over all actuarial survival rates were 57.1% at 5 year and 44.4% at 10 year in the earlier period, while it was 70.7% at 5 year in the later group. We concluded that retrograde cerebral perfusion allows resection of the dissected aorta including primary entry as well as open distal anastomosis, which contributes to the improvement of early and long-term results for acute type A aortic dissection. PMID- 9742794 TI - [Early and long-term outcome for surgical treatment of acute aortic dissection]. AB - This study was undertaken to determine the factors that influence the final outcome after the operation of acute aortic dissection. Twenty-one patients, the median age was 59 years (range 44 to 81), were operated at acute phase in 92 admitted into our hospital during the 13-year period between May 1985 and Jan. 1998. Preoperative complications included cardiac in 5 and ruptured with shock in 7, myocardial ischemia in 3 and stroke in 4. The ascending aortic reconstruction (9, 43%), ascending aorta and arch reconstruction (7, 33%) and other procedures (4, 19%) were performed using cardiopulmonary bypass with deep hypothermia and circulatory arrest or selective cerebral perfusion. The 30-day operative deaths were 6 (29%) and 5 (24%) late death occurred. Three out of 5 were aneurysm related deaths. The cause specific survival rates were 61% at 5 years and 51% at 8 years. The multivariably determined risk factors for death were as follows (p < 0.05): preoperative FDP; bleeding volume; postoperative renal complications; postoperative stroke. PMID- 9742796 TI - [Determinants of long-term outcome for operative survivors of acute aortic dissection]. AB - Predictability of aorta-related complications and survival was examined in 79 operative survivors of acute aortic dissection. Follow-up was 94.9% complete and totaled 458 patient-years. Actuarial survival was 93 +/- 3% (+/- S.E.) (n = 43) at 5 years, and 74 +/- 8% (n = 13) at 10 years. Survival was significantly lower in patients having neurological complication. Freedom from aorta-related complications was 82 +/- 5% (n = 37) at 5 years, and 67 +/- 8% (n = 11) at 10 years. Multivariate Cox regression analysis identified residual entry and leak on anastomotic site as independent predictors of aorta-related complications. We conclude that in the treatment of acute aortic dissections, reducing the incidence of residual entry and leak on anastomotic site improves long-term outcome. PMID- 9742797 TI - [Influence of acute aortic dissection on long-term results of aortic root reconstruction in patients with Marfan's syndrome]. AB - Objective of this study is to evaluate influence of acute aortic dissection on long-term results of aortic root reconstruction in patients with Marfan's syndrome. 19 patients who underwent consecutive aortic root reconstruction between 1985 May to 1998 February were retrospectively reviewed. Patients who associated acute aortic dissection at the time of operation (group D, n = 7) were compared long-term results with those who did not (group non-D, n = 12). Mean follow-up period was 5.1 +/- 3.2 years and longest follow-up term was 12.5 years. In each group early postoperative death was found in one patient. In-hospital mortality in each group were respectively; 14.2% and 8.3%. Late deaths were found in four patients who all belonged to group D, respectively caused by; rupture of thoracoabdominal aneurysm, DIC after subsequent thoracoabdominal surgery, sepsis due to prosthetic valve endocarditis, and sudden death. Actuarial overall survival rate including operative death in D and non-D group were respectively; 0.0 +/- 0% at 6.6 years and 91.1 +/- 8.0% at 12 years. Overall cumulative survival rate was 56.6 +/- 14.0%. Freedom from cardiovascular events, in D and non-D group were respectively; 0.0 +/- 0% at 6.6 years and 60.0 +/- 25.3%, and freedom free form dilatation of residual aorta were respectively; 0.0 +/- 0% at 6.6 years and 100.0 +/- 0% at 12 years. Freedom from subsequent cardiovascular surgery in group D, group non-D and over-all patients were respectively; 0.0 +/- 0% at 6.6 years, 60.0 +/- 25.3% at 12 years and 42.6 +/- 20.2%. In this study, acute aortic dissection in Marfan's syndrome significantly increased late cardiovascular events including dilatation of residual aorta, subsequent aortic surgery and late mortality. On the other hand, excellent long-term results after aortic root reconstruction were found in non-dissection Marfan's syndrome. Considering high incidence of late dilatation of residual aorta, simultaneous total arch replacement with aortic root reconstruction is recommended in acute dissecting Marfan's syndrome. Whereas, preventive simultaneous arch replacement is not required in non-dissecting Marfan's syndrome because of less postoperative vascular events. PMID- 9742798 TI - [Type A acute aortic dissection: late reoperations for dilatation of the distal false lumen and aortic regurgitation]. AB - From January 1990 to December 1996, 71 patients aged 33 to 79 years (mean 60 +/- 11) underwent an emergency operation for type A acute aortic dissection. Fifty three (74.6%) survived, and were followed 7 to 94 months (mean follow up 2.9 +/- 1.8 years) after the first operation. Five patients underwent reoperation for dilatation of the distal false lumen 7 to 52 months (mean period, 25 months) after primary repair. One patient underwent replacement of the arch and descending aorta, three patients underwent replacement of the descending aorta, and one patient underwent the stented graft implantation, resulting in closure of the entry site. Three patients underwent reoperation for severe aortic valve regurgitation 12 to 31 months (mean period, 24 months) after primary repair. Two patients underwent aortic valve replacement, and one patient underwent aortic root replacement. The actuarial freedom from reoperations was 81 +/- 6.9% at 3 years, and 73 +/- 9.9% at 5 years. PMID- 9742800 TI - [Long-term surgical results of acute aortic dissection]. AB - Between 1988 and 1997, 69 patients underwent surgery for acute aortic dissection: 65 patients had an acute type A (AcA) and 8 an acute type B (AcB) aortic dissection. The hospital mortalities were 16.9% for AcA and 25% for AcB. Actuarial survival rates including hospital deaths after 5 and 8 years were 70% and 58% for AcA patients and no late death or cardiovascular event occurred in AcB patients during follow-up periods. Freedom from cardiovascular events was 88% and 70% at 5 and 8 years, and freedom from reoperation was 94% at 5 and 8 years for operative survivors of AcA. There were no differences on actuarial survival rates between ascending aortic repair and arch repair, ringed-graft and suture anastomosis, and postoperatively patent false lumen and closed false lumen. There were no differences on freedom from cardiovascular events between ascending aortic repair and arch repair, and ringed-graft and suture anastomosis, however, patients with postoperatively patent false lumen showed significantly lower freedom from cardiovascular events registering 85% and 59% at 5 and 8 years compared with 94% at 5 and 8 years in patients with closed false lumen. PMID- 9742799 TI - [Postoperative follow-up of acute aortic dissection and the result of second operations]. AB - Between January 1990 and March 1998, surgical treatment was performed in 75 patients with acute aortic dissection at Omiya Medical Center. Seventy-three patients (97%) of them were classified type A aortic dissection. Thirteen of them died due to hemorrhage (5 cases), cardiac failure (3 cases), visceral ischemia (3 cases) and others (2 cases) after the operation. Hospital mortality rate was 17.3%. Sixty-two survivors were followed up to 8 years. During the follow up period, five patients died due to pneumonia, gastric cancer, cardiac failure, brain hemorrhage and unknown event. Five-year survival rate including hospital death was 77% evaluated by Kaplan-Meier method. Second operations for the enlargement of residual false lumen with the entry were performed in 4 patients (6.5%) of 62 survivors. Second operations were also performed in other 6 patients referred to us from other hospitals because of the enlargement of false lumen. Nine of them survived (90%) and returned to their daily life. PMID- 9742801 TI - [Long-term results of operation for acute type A aortic dissection]. AB - We report results of surgical treatment in 30 patients with acute type A aortic dissection. The average age of the 25 patients without the Marfan syndrome was 59.2 (range 51-76), and male/female ratio was 11/14. The average age of the five patients with the Marfan syndrome was 36.8 (range 27-48), and male/female ratio was 2/3. As an adjunct, we used deep hypothermic circulatory arrest during ascending aortic replacement, while selective cerebral perfusion was employed during aortic arch replacement. Operative procedures for the non-Marfan patients included 14 ascending aortic replacement and 11 ascending and aortic arch replacement, while the Marfan patients underwent extensive aortic replacement that included three aortic arch replacement combined with the Bentall operation, one extensive replacement from the ascending aorta to the descending thoracic aorta and one ascending and aortic arch replacement. One patient died early after the operation and the early mortality rate was 3.3%. No patient developed new brain complication related to the operation. During the follow-up period, three patients died and two patients required a total of three subsequent distal operations. Cumulative survival rate was 89% at one year, 85% at three years, 85% at five years. Cumulative cardiovascular event-free rate was 89% at one year, 85% at three years, 77% at five years. Early and long-term results of surgical treatment for acute type A aortic dissection was satisfactory. This seems to result from the use of deep hypothermic circulatory arrest and selective cerebral perfusion as an adjunct and application of aortic arch replacement when the aortic arch is dilated or intimal tear is located in the aortic arch or more distally. PMID- 9742803 TI - [The long-term results of surgical treatment for acute type A aortic dissection]. AB - Between 1992 and 1996, 31 patients with acute type A aortic dissection underwent the surgical treatment on an emergency basis. The overall early mortality rate was 12.3% (4 patients), and the overall 5-year survival rate (calculated by Kaplan-Meier method) was 76.9% respectively. Although there is controversy about the operative procedure whether the aortic arch should be replaced or not in aortic arch dissection in which the primary intimal tear is located at or extends into the arch, the resection of the aortic arch tear and simultaneous arch repair with graft replacement of the ascending aorta would be the acceptable procedure in the long term period with the recent improvement of adjunct technique for the prevention of cerebral ischemia and heart protection. The composite graft replacement of the aortic root is also indicated in patients with acute dissection associated with annuloaortic ectasia or Marfan's syndrome and severely destroyed aortic root with the extension of dissection to the aortic root with acceptable mortality and survival rate in the early and late period. PMID- 9742802 TI - [Long-term results of patients undergone surgical repair for acute type A dissection]. AB - We evaluated the long-term results of 28 operative survivors who underwent surgical repair for acute type A dissection. Residual dissection of the thoracic aorta was found in 21 patients (75.0%) postoperatively, and 6 of them underwent reoperation with no operative death. There were 2 late death; one died in the third operation for thoracoabdominal aorta and the other died of cancer. Two of 4 patients with Marfan's syndrome required total aortic replacement ultimately. Since the elective reoperation can be performed safely, careful follow-up and management is essential for the improved survival of the patients with residual aortic dissection. PMID- 9742804 TI - [Late results of emergency operation in acute type A dissection]. AB - 77 cases of acute type A dissection underwent emergency operation during 5 years were studied. There were 37 men and 40 women, and 58 cases in type I, 19 cases in type II on DeBakey type. The grafting for the aorta including the tear was basically performed under selective brain perfusion and deep hypothermic circulatory arrest. The overall hospital mortality rate was 24.7%. However, it was 14% after using GRF glue. The overall actuarial survival rate for 58 patients who survived operation was 87% at 5 years after surgery. Late remnant false lumen related complications were recognized in four patients (6.9%). For two of them, subsequent operations were performed. Another two patients died of rupture of remnant false lumen during observation. Aortic valve regurgitation was recognized preoperatively in 13 patients. None of them required subsequent operation in late phase. Results and countermeasure for late remnant false lumen-related and aortic valve-related complications were investigated. PMID- 9742805 TI - [Emergent operation for acute Stanford type A aortic dissection]. AB - Results of emergent surgery for acute Stanford type A aortic dissection are still poor. We reviewed our cases to find out how we could ameliorate surgical results. In the past five years, 36 patients with type A aortic dissection underwent emergent surgery. The operation was started within six hours from the onset of symptoms in 16 patients. The extent of aortic replacement was just beyond the entry-bearing portion. The operation was performed with the aid of open distal anastomosis and selective cerebral perfusion. Operative death occurred in one patient. Postoperative complications developed frequently. Respiratory failure, renal dysfunction and low cardiac output syndrome were most frequent. Their co risk factor was perioperative bleeding which was related to young age, long operation time, arch replacement, additional procedure and failure of the entry site resection. Some modification of the suturing method might reduce hemorrhage. Prompt surgery following the diagnosis and appropriate selection of surgical tactics and anastomosis technique in order to reduce bleeding is essential to improve surgical results of acute type A aortic dissection. PMID- 9742806 TI - [Snare injury in coronary artery bypass grafting on the beating heart: report of 2 cases]. AB - From June to November 1997, 8 patients underwent coronary artery bypass grafting on the beating heart. The first 2 patients had snare injury. Postoperative angiography showed coronary stenosis distal to the LITA-LAD anastomosis in the first case and a pseudo-aneurysm at the septal branch of the LAD in the second case, most likely due to snaring maneuver with 3-0 prolene or 3-0 GORE-TEX suture with a sharp needle. Since then, we have been satisfactorily utilizing the RETRACT-O-TAPE (a silicone tape with blunt needle, QUEST Medical, Inc.) for coronary occlusion to avoid injury to the native vessels and their branches. PMID- 9742807 TI - [An operative case of spontaneous rupture of the esophagus 10 days after the onset]. AB - This paper presents a rare case of spontaneous rupture of the esophagus operated on 10 days subsequent to its onset. A 69-year-old male, who was diagnosed as acute pancreatitis, came to this department after 10 days of conservative therapy. Emergency examination including computed tomography, esophagoscopy and esophagography indicated spontaneous rupture of the esophagus. At operation, despite severe inflammation of the pleural cavity, a 2 cm horizontal tear at the left wall of the lower esophagus could be directly closed, and reinforced with fibrin glue. Postoperative decompression therapy prevented the rupture of the closure. The early symptoms resemble other emergency diseases, thus making correct diagnosis difficult. Early management is required for lifesaving, and preoperative aggressive exploration must thus be conducted. Postoperative management including through decompression of the gastrointestinal tract is also essential, regardless of the mode of operation. PMID- 9742809 TI - [A case of surgically resected mediastinal mass histologically proven to be a mass-like tissue presenting extramedullary hematopoiesis accompanied by thalassemia]. AB - We experienced with a case showing a mediastinal mass on chest X-ray accompanied by thalassemia. Preoperative diagnosis for this mass was a neulogenic tumor based on the findings of roentgenogram. Surgical resection of the mass was performed, and histology revealed that it was a mass-like tissue presenting extramedullary hematopoiesis. Generally speaking, any tissues functioning as extramedullary hematopoiesis should not be resected in patients with anemia or showing bone marrow suppression. And it is also said that 111In-chloride scintigram is highly diagnostic for patients with a tissue functioning as extramedullary hematopoiesis. We had not performed 111In-chloride scintigram for this case preoperatively and performed a resection of the mediastinal mass carelessly. Care should be taken not to perform surgical resection carelessly in cases with a mediastinal mass accompanied by anemia or bone marrow suppression. PMID- 9742808 TI - [A case report of asymptomatical large carinal bronchial cyst]. AB - A 36-year-old man with a mediastinal bronchial cyst was reported. CT showed a mass with water density in the posterior mediastinum. MRI revealed that this mediastinal mass had high signal intensity on both T1-weighted and T2-weighted images. MRI was considered to be useful for the qualitative diagnosis of mediastinal tumors as cystic or solid. An operation was performed and histological diagnosis was bronchial cyst. Intracystic fluid study revealed high titer of total protein, amylase and tumor markers, low titer of glucose and LDH. PMID- 9742810 TI - [A case report of pulmonary actinomycosis presenting as a mass shadow on chest X ray]. AB - A 58-year-old man admitted to hospital because of hemoptysis. Chest X-ray showed a large mass in the right middle lobe. A tumor marker CYFRA was slightly elevated. Despite a detailed examination after admission, no definite diagnosis was made. Lung cancer was suspected and a middle lobectomy was performed. Histopathological specimen of resected lung showed typical "sulfur granule" of actinomycosis. Pulmonary actinomycosis should be included in the differential diagnosis of a pulmonary mass lesion. PMID- 9742811 TI - [A case of severe cytomegalovirus infection after the repair of coarctation of aorta]. AB - We report a 2-month-old boy without any immuno-compromised diseases, who suffered from the severe cytomegalovirus (CMV) infection after the subclavian flap aortoplasty and pulmonary artery banding for coarctation complex. He underwent the operation at 2 months old and received 2 units of irradiated packed red blood cells before and after the surgery. His postoperative course was uneventful but the interstitial pneumonitis, until he developed watery diarrhea 10 days after the surgery following hepatitis with the marked hepatomegaly 3 weeks after. Since CMV infection was confirmed as the cause of the pneumonitis, enterocolitis and hepatitis, he was initially treated by gamma-globulin with the high CMV titer at a dose of 200 mg/kg/day for 2 days and ganciclovir at a dose of 10 mg/kg/day for 14 days. Because of the persistent CMV infection, he needed two more treatments of ganciclovir at the same dosage and gamma-globulin once a week for 2 months. He finally recovered from severe CMV infection 5 months after the above treatments. In conclusion, the severe CMV infection can occur by blood transfusion even in the surgical case with normal immune system. If one finds pneumonitis, hepatitis or enterocolitis after any type of surgery with history of blood transfusion, CMV infection should be suspected as the cause of these diseases. PMID- 9742812 TI - [A surgical case of angina pectoris with a severe stenosis of Lt. mid-cerebral artery: the usefulness of the monitoring of cerebral blood flow]. AB - A 64-year-old male patient had two episodes of transient ischemic attack and a cerebral infarction. Cerebral angiography showed 50% stenosis at the junction of left internal carotid artery and 90% stenosis at left mid-cerebral artery (MCA). Coronary angiography showed two vessel disease with arteriosclerotic change and underwent coronary artery bypass grafting. To prevent intraoperative cerebral infarction, we used brain protect solution just before starting ECC, set perfusion flow around 3 l/min/m2, monitored the flow of left MCA using Transcranial Doppler (TCD) and the saturation of left internal jugular vein (SjO2) continuously. PaCO2 was controlled around 45 mmHg. TCD showed good pulsatile flow, and SjO2 was kept over 60%. The patient recovered consciousness 2 hours after operation in the intensive care unit without paresthesia. We thought the number of open-heart cases with cerebrovascular disease increased, and pulsatile low of ECC by intraaortic balloon pumping and the monitoring of SjO2 are useful for the cases. PMID- 9742813 TI - [A voice-controlled robot assisted thoracoscopic surgery for pulmonary tumor]. AB - We herein report a case of voice-controlled robot assisted thoracoscopic surgery for a right apical tumor in a 55-year-old male. The patient was referred to our hospital for further examination of an abnormal shadow on chest X-ray. A chest computed tomographic scan indicated a suspicion of malignancy. Thoracoscopic surgery using a voice-controlled robot was conducted. The thoracoscope connected with the robotic arm was inserted through the trocar. In all maneuvers (forward and backward, right and left, up and down, insertion and extraction), the robotic arm moved appropriately immediately after the voice commands were issued, and favorable operative fields were obtained. Thoracoscopic partial resection of the involved lung was successfully performed with no complications. The operative time was 140 min. The postoperative course was uneventful. Voice-controlled robotic technology innovations may become the operating surgeon's third arm in the future. They will be further used not only in laparoscopic but also in thoracoscopic surgical procedures. Regular use will lead to greater skill in thoracoscopic application with the advent of new robotically assisted endoscopic surgery. PMID- 9742814 TI - [Arrhythmia and hemodynamic management during coronary artery bypass grafting without cardiopulmonary bypass]. AB - We have presented a method by which perfusion can be maintained to the coronary artery while surgeon sutures anastomosis in undergoing coronary artery bypass grafting without cardiopulmonary bypass. The shunt from femoral artery to right coronary artery (RCA) worked well in preventing the onset of severe arrhythmia and hypotension secondary to occlusion of the RCA. PMID- 9742815 TI - [Autologous blood donation with recombinant human erythropoietin prior to elective cardiac surgery in anemic patients]. AB - Despite the reducing exposure to allogeneic blood in cardiac surgery, most of patients with anemia still require allogeneic blood. In this study, we have attempted to harvest the blood from cardiac patients with baseline hemoglobin levels below 11.0 g/dl using recombinant human erythropoietin (rHuEPO). 29 anemic patients undergoing cardiac surgery at our hospital between January 1994 and March 1997 were divided into two groups: 3 weeks' treatment with recombinant human erythropoietin (rHuEPO) and blood donation (group 1, n = 15) and iron supplementation alone (group 2, n = 14). There were no statistically significant differences among the two groups in patients characteristic and surgical data. No serious adverse events after phlebotomy were apparent in patients donating autologous blood. Patients in group 1 had significantly higher hemoglobin levels than patients in group 2 at 7 days before operation. The number of reticulocytes were increased at just before operation in group 1, whereas group 2 showed no significant increase. The estimated hemoglobin increase in group 1 were higher at 7 days and just before operation. In 75% of group 1, allogeneic blood transfusion could be avoided, while all patients in group 2 received allogeneic blood transfusion. This study suggests that the combination of rHuEPO administration and autologous blood donation would be beneficial for anemic patients in elective cardiac surgery. The use of rHuEPO should not be restricted to anemic patients. PMID- 9742816 TI - [Management for the postoperative mediastinitis in infancy]. AB - Postoperative mediastinitis is a rare but life-threatening complication after cardiac surgery. We successfully managed three infants with postoperative mediastinitis. When the postoperative mediastinitis was suspected, intravenous infusion of antibiotics (Vancomycin) and local irrigation were performed. The reoperation for closure was planned when the value of c-reactive protein decreased to 1.0-2.0. An application of a pectoral musculocutaneous flap was effective when the sternum was destroyed by infection. PMID- 9742817 TI - [Long-term follow-up of patients with small St. Jude aortic valve prostheses]. AB - Thirty-five patients received small sized 19 or 21 mm valves (group I) and 53 patients received 23 or 25 mm valves (group II). At follow-up a significant reduction in the left ventricular mass was found for both patient groups (p < 0.0001). The patients in group I had higher peak pressure gradients than did those in group II. However, there was no significant difference between the two groups, or any significant correlation between peak pressure gradients and body surface area. Actuarial survival was 84.7% at 15 years for group I and 85.9% at 17 years for group II. Actuarial freedom from valve-related events was 91.4% at 15 years for group I and 82.7% at 17 years for group II. There was no significant difference in survival or valve-related event free curves between the two groups. After implantation of small SIM valves, significant left ventricular mass regression was obtained and the results were comparable to those for valves of other sizes. The long-term performance of aortic valve replacement with small valves was satisfactory as judged by improvement in the functional class of patients, survival statistics, the durability of the prosthesis, and valve related morbidity comparable to that of valves of other sizes. PMID- 9742819 TI - [Fenestration of diaphragm for management of persistent postoperative pleural effusion: clinical experience and experiment in rabbits]. AB - The management of persistent postoperative pleural effusion is still considered difficult. We report here our experience with such a case, successfully managed with fenestration of the diaphragm-the first of it's kind. Clinical Experience: A two-year-old boy with double outlet right ventricle, underwent right heart bypass procedure. Due to low output and high venous pressure, he was on ventilator until the 26th postoperative day. The pericardial and right pleural effusion persisted till the 60th postoperative day. Right phrenic nerve palsy and atelectasis of right lower lobe were suspected to contribute to it. We performed a plication of the right diaphragm and fenestration of the pericardium and right diaphragm. A T shaped incision was made on the right diaphragm and the edges were trimmed and strengthened with non-absorbable suture into a circular shaped defect of 1.5 cm diameter. The defect was closed with a Dacron mesh allowing passage of fluid across. Pleural effusion decreased immediately and he was discharged a month after the procedure. Experimental Study: The above procedure was experimented in rabbits in whom a contrast medium injected into the pleural cavity could easily drain into the peritoneum through the fenestation, proved by fluoroscopy. CONCLUSION: Fenestration of the diaphragm is an effective procedure to manage persistent pleural effusion. PMID- 9742818 TI - [A case of pulmonary epithelioid hemangioendothelioma diagnosed by video-assisted thoracoscopic biopsy]. AB - A regular annual physical checkup for a 46-year-old man revealed multiple small nodules, which were less than 1 cm in diameter, in both lungs. Metastatic lung tumors were suspected, but no primary lesion was found in the general examination. Video-assisted thoracoscopic surgery (VATS) diagnosed multiple pulmonary epithelioid hemangioendothelioma (PEH), so no treatment was performed. Careful follow-up showed no significant change during 10 months. VATS is thus a certain, noninvasive procedure for the diagnosis of PEH. PMID- 9742820 TI - [Successful removal of an infected pacemaker electrode adhered to the tricuspid valve]. AB - A 77-year-old male suffered from pacemaker lead infection with pacing failure. Because he had a severe fever, we performed an interventional removal procedure with the help of a lead removal kit. However, the tip of the lead could not be withdrawn via the right internal jugular vein because it adhered tightly to the tricuspid valve. Two days later, we proceeded with an open removal procedure under cardiopulmonary bypass. The lead could be removed without any complication, and no inflammation was observed postoperatively. We report the case and discuss the indications and limitations of both the interventional and open methods. PMID- 9742821 TI - [Successful surgical treatment for severe mitral valve annulus enlargement and mitral regurgitation 5 years after aortic root replacement in Marfan syndrome: a case report]. AB - Major cardiovascular lesions associated with Marfan syndrome include aortic diseases such as aortic root dilatation and mitral disease. If untreated, cardiovascular manifestations of the Marfan syndrome cause death in one half of patients during the first four decades of life. Although aortic diseases in the Marfan syndrome are responsible for most of serious morbidity and mortality, 60 80% of patients with Marfan syndrome have mitral valve dysfunction. The results of surgical treatment for aortic diseases in Marfan patients have recently shown significant improvement. However, there is no acceptable consensus about the timing and the technique of surgical intervention for mitral diseases. Especially mitral valve diseases is the most common cause of morbidity and mortality in infants with the Marfan syndrome. We report herein a case of Marfan syndrome (21 year-old female) who had undergone two operations for aortic root dilatation five years before she underwent the surgical treatment for progressive heart failure due to severe mitral annulus enlargement and mitral regurgitation. PMID- 9742822 TI - [A case report of aortic regurgitation associated with rheumatic arthritis]. AB - We report a rare case of aortic regurgitation (AR) associated with rheumatic arthritis (RA). A 61-year-old female was admitted to our hospital with severe heart failure due to AR. She had a 8-year history of RA and had been treated with steroid therapy. Trans-esophageal echocardiography showed thickness and shortening of non coronary cusp (NCC) of aortic valve. After treated for heart failure, aortic valve replacement with SJM-HP 17 mm was done. At operation, right and left coronary cusps were almost normal, but NCC was thickened and shortened. The valve of NCC histology showed granuloma. She recovered uneventfully. Two cases of cardiac valvular disease, associated with granuloma of RA, were reported in Japan. We discussed these cases in this paper. PMID- 9742823 TI - [A surgical case of cardiac malignant lymphoma in right atrium]. AB - We reported a case of malignant lymphoma originating from the right atrium. The patient was a 71-year-old man who had no symptoms associated with heart failure and arrhythmias. At the time of admission the patient was suggested the presence of a tumor in the right atrium by an echocardiogram accidentally. Coronary angiography revealed a feeding artery to a tumor. Although cytological confirmation was not obtained, diagnostic extirpation of tumor was performed under cardiopulmonary bypass. The histological diagnosis was malignant lymphoma of B-cell origin. His postoperative course was uneventful and no recurrence had been observed one year postoperatively without chemotherapy. PMID- 9742824 TI - [An experience of video-assisted thoracoscopic surgery for an intrapulmonary needle under CT-guided marking]. AB - A 46-year-old female visited a near hospital, complaining of repeating skin eruption with spontaneous remission. A chest X-ray and CT examinations revealed a needle in the left S6, about 5 mm in length, which was suspected of causing the eruption. She was admitted to our hospital for operation. After CT-guided marking, video-assisted thoracoscopic surgery was performed. Because the needle was too small to be palpable, we decided to respect of it with the lung surrounding the marker and clamped the lung, followed by finding the needle finally. The analysis of the removed needle showed that it contained lead and tin besides iron, nickel and chromium, which are components of stainless steel. Although dermatological examination could not reveal the relation between the needle and the eruption, it has never recurred since the operation. It is thought that CT-guided marking is very useful for resection of an intrapulmonary needle and that a foreign body should be removed for the possibility of being harmful. PMID- 9742825 TI - [A case report of dumbbell neurogenic tumor of the superior mediastinum]. AB - A 15-year-old female was admitted because of the superior mediastinum mass on chest X-rays and sensory loss of ulnar side of the left arm. Computed tomographic scanning and magnetic resonance imaging revealed that the tumor was dumbbell shaped and invaded the vertebral canal through the intervertebral foramen between C 7 and Th 1. At first laminectomy of vertebrae (C 6-Th 1) was made in a prone position and intra-spinal portion of the tumor was resected. Then the patient was placed in a supine position and the chest was opened by left hemicollar incision and sternotomy to the 2nd intercostal space. The tumor was divided into two parts at the level of 1st rib and completely removed. The pathological diagnosis was schwannoma. This procedure is safe and useful for dumbbell type tumor located in superior mediastinum, especially in case of large tumor from neck to the thoracic cavity. PMID- 9742826 TI - [Successful video-assisted thoracoscopic bullectomy and Nd-YAG laser ablation]. AB - We report here two cases of giant/multiple emphysematous bullae treated with video-assisted thoracoscopy. The first case was a 35-year-old male who was referred to our hospital because an abnormal shadow was casually pointed out in a chest roentgenogram. Chest computed tomographic scan showed giant bulla in the left upper lobe. The second patient was a 37-year-old male who had a symptom of shortness of breath on exertion. Chest roentgenogram and computed tomographic scan revealed multiple bullae at bilateral apical regions. They were performed resection of bulla with stapler and Nd-YAG laser ablation to the emphysematous surroundings using video-assisted thoracoscopy. They had recovered sufficiently to be discharged from our hospital. We think Nd-YAG laser ablation is effective to prevent postoperative air leakage and recurrence of bullae because it makes surrounding tissue tight and continuous. PMID- 9742827 TI - [A case of panpleuropneumonectomy for diffuse pleural mesothelioma]. AB - A 73-year-old male was admitted with dyspnea and cough. The chest X-ray showed left massive pleural effusion and diffuse pleural tumor in the left thorax. It was diagnosed as epithelial-cell type mesothelioma by pleural needle biopsy. After conforming the regression of the tumor from conducting two courses of combined treatment with cisplatin and doxorubicin, panpleuropneumonectomy was performed. He died from sepsis on the thirty second day after operation due to complication of postoperative diaphragmatic hernia and gastric perforation. When conducting a panpleuropneumonectomy to diffuse pleural mesothelioma, the most appropriate approach must be taken to the combined with resection and reconstruction of the diaphragma. PMID- 9742828 TI - [A case of treatment with Dumon stent for tracheal stenosis due to lung cancer with hypercalcemia]. AB - A 65-year-old man was facing to tracheal stenosis due to proximally developed lung cancer with hypercalcemia mediated by production of PTHrP from the cancer cells. We treated the stenosis use of Dumon stent and hypercalcemia with drugs such as biphosphonate and so on. The treatment was effectively keep the patient under better performance status for about 3.5 months. PMID- 9742829 TI - [Three resected cases of pulmonary cryptococcosis]. AB - Case 1 was a 53-year-old female who had a small nodule in the right S3 segment on chest CT. As she was not diagnosed by transbronchial lung biopsy (TBLB), open thoracotomy was performed. Case 2 was a 65-year-old female who had a nodule with pleural indentation in the right S6 segment. As this nodule showed difficulty to differentiate from small lung carcinoma, thoracoscopic surgery was performed. Case 3 was a 63-year-old female who had multiple lesions with cavity in the left S4 and S5 segments, which was preoperatively diagnosed by TBLB. She was performed thoracoscopic partial resection of the lingular segment because of poor response to antimycotic agents. All cases received preventive antimycotic agents for one or two months after the operation. There was no recurrence or postoperative meningitis. Thoracoscopic surgery is the effective procedure for the diagnosis and treatment of the localized pulmonary cryptococcosis. PMID- 9742830 TI - [Reduced breast cancer mortality. Early treatment, as endocrine and cytostatic adjuvant therapy is effective]. PMID- 9742832 TI - [Single chromosome deviations can determine the effect of drugs]. PMID- 9742831 TI - [Age itself is not a contraindication to surgery. New minimally invasive surgical procedures reduce the risks]. PMID- 9742833 TI - [Stroke after thoracic surgery. An unusual hypothermic method reduces the number of cases]. PMID- 9742834 TI - [New arguments against euthanasia?]. PMID- 9742835 TI - [Does careful documentation solve problems with certificates?]. PMID- 9742836 TI - [Health insurance authorities should take the responsibility!]. PMID- 9742837 TI - [How to update drug information]. PMID- 9742838 TI - [The employer should pay for personnel with burnout symptoms]. PMID- 9742839 TI - [Freedom for and against circumcision!]. PMID- 9742840 TI - [Laryngospasm--a rough consideration?]. PMID- 9742841 TI - [Is the registration of drug prescriptions of some benefit?]. PMID- 9742842 TI - [Not only "feelings" behind syndromes connected to amalgam and electricity]. PMID- 9742843 TI - [Syphilis--500-year anniversary of a dreaded plague]. PMID- 9742844 TI - [Eros's illness. A complex net of abortions, STD and contraceptive agents]. PMID- 9742845 TI - [Which patients need gallstone surgery?]. AB - Although gallstone disease is common, few patients develop serious complications. Recent decades have witnessed manifest changes in the frequency of gallstone surgery in Sweden. Although variation in the prevalence of gallstone disease may be one explanation, uncertain indications for gallstone surgery may also be a contributory determinant of the varying cholecystectomy rates. Findings in epidemiological and clinical studies suggest that cholecystectomy should be performed in cases of frequent gallstone symptoms or acute gallstone complications, whereas expectant management is to be recommended in cases of infrequent symptoms or where their link with gallstone problems is in doubt. PMID- 9742846 TI - [A severe adverse effect of penicillin on the liver. Drug insurance covered the compensation]. PMID- 9742848 TI - [A two-year old girl lost her hair--gluten intolerance was the cause]. PMID- 9742849 TI - [Echocardiography in acute pulmonary embolism. Not a routine method but useful in the diagnosis of simultaneous hemodynamic disorders]. AB - Echocardiographic diagnosis of acute pulmonary embolism as illustrated by three case reports is discussed in the article. Acute pulmonary embolism was diagnosed by demonstration of right heart strain in one case, of long vermiform thrombi floating in the right atrium in another, and in the third case by demonstration of a long thrombus lodged in the foramen ovale, astride the atrial septum, and with its ends floating in either atrium. Thus, as echocardiography enables pulmonary embolism to be diagnosed by demonstration either of right heart strain or of intracardial thrombi, it is a useful diagnostic tool in cases of haemodynamic compromise, though it does not detect minor pulmonary embolism. PMID- 9742847 TI - [Hyponatremia as a side effect of serotonin uptake inhibitors]. AB - Hyponatraemia is a possible, potentially serious adverse reaction to treatment with selected serotonin re-uptake inhibitors (SSRIs). The article consists in a review of the 27 cases of such reactions that have been reported to the Swedish Medical Products Agency. The data from these reports suggest the risk of hyponatraemia to be particularly manifest during the first few weeks of treatment, and to be greater in women, the elderly, and patients concomitantly treated with diuretics. In the event of vague, non-specific symptoms occurring in conjunction with SSRI treatment, measurement of the serum sodium concentration is recommended. PMID- 9742850 TI - [Fragile X chromosome--a male weakness]. PMID- 9742851 TI - [Early diagnosis of parenchymal lung diseases is now possible. Reduced risk of fibrosis and irreversibly reduced lung function]. AB - Although parenchymal, or interstitial lung diseases constitute a heterogeneous category, a common feature is the occurrence of an alveolar and interstitial inflammatory reaction. The alveolitis may regress, but may also heal with scarring and fibrosis, resulting in chronic impairment of lung function. Early diagnosis is important in order to identify patients at risk of lung function impairment. Diagnosis has been facilitated by the advent of sophisticated, particularly radiological and bronchoscopic methods in recent years, thus enabling intervention at an earlier stage in the disease course. The various diagnostic alternatives available for the investigation of parenchymal lung diseases are reviewed in the article. PMID- 9742852 TI - [Significant regional differences in the frequency of vascular surgery]. AB - The six health care regions of Sweden were compared with regard to the frequency of vascular surgery for three diagnoses: chronic lower extremity ischaemia, abdominal aorta aneurysm, and carotid stenosis. In 1995, the frequency of intervention for chronic lower extremity ischaemia varied from 26/100,000 of the population in northern Sweden to 68/100,000 in the southern region, the variation being greater for critical limb ischaemia than for intermittent claudication. In the country as a whole, the frequency of abdominal aorta aneurysm surgery increased five-fold from 1987-89 to 1993-95. During 1995, regional figures varied from 4.7 to 8.4 per 100,000 for elective procedures, and from 3.8 to 5.5 per 100,000 for emergency procedures. Overall surgical mortality varied regionally, and emergency surgery mortality differed between regional and county hospitals. Carotid surgery manifested the greatest regional difference in frequency, which was 7-fold greater in the southern than in the northern region, while its overall mean frequency was 6/100,000. PMID- 9742853 TI - [Colonic-cutaneous fistula after colon perforation. Complication after percutaneous endoscopic gastrostomy]. PMID- 9742854 TI - [A proposal from the EU makes the development of "orphan drugs" easier]. PMID- 9742855 TI - [Doctor Orfila from Menorca as Strindberg's secret patron saint]. PMID- 9742856 TI - [Growth factors in the process of inflammation and fibrosis in the lung]. AB - Growth factors are known not only to cause a mitogenic response and alter differentiated characteristics of the target cells, but also to play important roles in intercellular signaling. Many growth factors are expressed in the embryonic and regulate embryogenesis. Pulmonary fibrosis is characterized by a complex process involving chronic inflammatory reaction, fibroblast proliferation, and abnormal deposition of interstitial collagen as a result of excess healing reaction. In the early phases, TNF-alpha, IL-beta and GM-CSF secreted by alveolar macrophages regulate and enhance pulmonary inflammation. On the contrary, TGF-alpha, KGF and HGF have been reported to enhance repair of alveolar epithelium and vascular endothelium in the injured lung. Furthermore, growth factors produced by alveolar macrophages and epithelium, such as PDGF, TGF beta and activin A and belongs to the TGF-beta supergene family are known to play cardinal roles in fibroblast proliferation and pulmonary fibrosis. Further works concerning this complex growth factors (cytokines) network are required to provide a basis of the pathophysiology of pulmonary fibrosis. PMID- 9742857 TI - [Effectiveness of continuous intravenous prostaglandin E 1 against pulmonary hypertension]. AB - Pulmonary hypertension is a progressive disease for which no effective therapy has been found. Short-team treatment with alprostadil alfadex (PGE 1) can improve hemodynamics but the effects of long-term treatment have not been reported. We administered intravenous PGE 1 to 16 patients with pulmonary hypertension for 4 weeks in addition to conventional therapy. Hemodynamic and symptomatic effects were evaluated. The infusion was interrupted in 7 patients (44%) because of fever and an increase in levels of C-reactive protein in serum. In the remaining 9 patients, the drug lowered pulmonary artery pressure and resistance, and imareased cardiac output significantly in 7 patients. PGE 1 can improve hemodynamics and relieve symptoms in patients with pulmonary hypertension, barring severe side effects. PMID- 9742858 TI - [Effect of salbutamol on proliferation of human bronchial epithelial cells: role of MAP kinase]. AB - To determine whether stimulation of beta-adrenoceptors affects proliferation of airway epithelial cells and, if so, whether activation of mitogen-activated protein kinase (MAPK) is involved, we studied cultured human bronchial epithelial (16-HBE) cells in vitro. The 16-HBE cells were grown to subconfluence in 96-well plates, and their growth was inhibited by incubation in serum-free medium for 72 h. The cells were ten incubated in the presence of saltbutamol (SAL, 10(-7) M), a specific beta(2)-adrenoceptor agonist. Proliferation of the cells was evaluated by MTT assay and total DNA content, and activation of MAPK was assessed by immunocytochemistry and Western blotting for phosphorylated MAPK (phospho-MAPK). Immunocytochemistry and immunoblots demonstrated that phospho-MAPK was expressed within minutes of SAL exposure. This effect of SAL was as potent as that of 10% serum, and was greatly inhibited by treatment with propranolol. These results suggest that SAL is a potent mitogen of airway epithelial cells and that its effect may be exerted by beta(2)-adrenocepter-mediated activation of MAPK. PMID- 9742859 TI - [Relaxation of canine pulmonary arteries caused by stimulation of atypical beta adrenergic receptors]. AB - To examine possible contributions of beta(3)-adrenoceptors to catecholamine induced pulmonary vasodilation, we studied isolated canine pulmonary arterial segments under isometric conditions. Addition of beta-adrenoceptor agonists produced a concentration-dependent relaxation of tissues precontracted with 50 mM KCl; the rank order of potency was isoproterenol (ISO, 1) > salbutamol (SAL, 0.97) > selective beta(3)-adrenoceptor agonists CL 316243 (CL, 0.87) and BRL 37344 (BRL, 0.86). Relaxant responses to SAL were competitively antagonized by the beta(2)-adrenoceptor antagonist ICI 118551 and the pA2 value was 6.67 +/- 0.21 (mean +/- SE), whereas the response to CL was weekly antagonized only by a high concentration of ICI 118551 (10(-5) M) and the apparent pA2 value was 5.24 when alpha-and beta(1)-adrenergic receptors were blocked. By contrast, the atypical beta-adrenoceptor antagonist cyanopindolol antagonized CL-induced relaxation in a competitive manner; the pA2 value was 6.71 +/- 0.12, which was lower than that with salbutamol (p < 0.05). Intracellular cyclic AMP levels were increased in a contraction-dependent manner by CL. These results suggest that beta(3)-adrenoceptors may exist in canine pulmonary arterial smooth muscle and that stimulation of this atypical receptor causes vasodilation through a cyclic AMP-dependent pathway. PMID- 9742860 TI - [Emergency room visits by patients with exacerbations of asthma]. AB - We retrospectively analyzed patterns of emergency room visits by patients with exacerbations of asthma from December 1995 through November 1996. A total of 591 episodes in 198 patients were reviewed. The average age was 35.8 years, ranging from 15 to 71. The largest number of visits occurred in September. The number of visits per year ranged from 1 to 22; the mean was 2.9 per year. In patients who were followed on a regular basis at our institution, serve attacks accounted for 7.1% of the total, compared with 21.6 percent at other hospitals or outpatient clinics. We suspect that this difference was related to differences in the use of inhaled steroids. At our institution, 89% of patients were taking inhaled steroids; at other hospitals or clinics, only 21% were taking inhaled steroids. Of the 198 patients, 33 fulfilled one of the following criteria: (1) endotracheal intubation for respiratory failure or respiratory arrest, (2) respiratory acidosis (pH < 7.35) without endotracheal intubation; 27% of those patients had been given a diagnosis of mild asthma before the acute exacerbation. We conclude that patient education and standard guidelines for treatment of asthma, are very important for appropriate management of asthma, to prevent exacerbations and asthma-related deaths. PMID- 9742861 TI - [Diffuse aspiration bronchiolitis in an elderly woman]. AB - A 74-year-old woman was admitted to the hospital because of persistent nocturnal coughing and abnormal shadows on chest x-ray films. The films showed cavities in the right upper lobe and small nodular shadows in the right upper, lower, and left upper lung fields. Examination of a specimen obtained by transbronchial lung biopsy showed nonspecific inflammatory changes. An open-lung biopsy was done. Histopathological examination showed evidence of diffuse aspiration bronchiolitis and actinomyces. Actinomyces is a member of the endogenous flora of the oral mucous membranes. Our diagnosis was diffuse aspiration bronchiolitis caused by micro-aspiration of oro-pharyngeal secretions during sleep. PMID- 9742862 TI - [Isoniazid-induced pneumonitis]. AB - An 82-year-old man was treated with isoniazid (INH) because of a low-grade fever. On the 9th day of treatment, dry coughing and general malaise developed. On the 30th day, he was admitted to our hospital. A chest-X ray film showed infiltrative shadows in the right middle and lower lung fields, but a chest CT scan showed an abnormal lung density in the right lower lobe. Abnormal laboratory findings included leucocytosis, liver dysfunction, hypoxemia, low vital capacity, low diffusing capacity and a high level of C-reactive protein. A differential cell count of the bronchoalveolar lavage fluid (BALF) showed many neutrophils and lymphocytes; examination of a specimen obtained by transbronchial lung biopsy (TBLB) revealed edema of alveolar walls, lymphocyte infiltration, and proliferation of type II alveolar epithelial cells. A drug lymphocyte stimulation test (DLST) against INH was positive. After discontinuation of INH, symptoms resolved, laboratory findings became normal, and the infiltrative shadows in the right middle and lower lung fields disappeared. The clinical course and the findings of BALF, TBLB, and DLST suggested the diagnosis of pneumonitis caused by INH. PMID- 9742863 TI - [Diffuse panbronchiolitis with myeloperoxidase-specific antineutrophil cytoplasmic antibody-related vasculitis]. AB - A 46-year-old woman was referred to our department in July 1996 with complaints of fever and myalgia in her calves. She had a 20-year history of purulent sputum; diffuse panbronchiolitis had been diagnosed in 1983. Physical examination revealed low-pithed rhonchi over the lung fieldis and hypesthesia of the right leg. She had a white blood cell count of 16,100/mm3, including 4% eosinophils, and a platelet count of 80.0 x 10(4)/mm3. The serum IgE level was 2,200 U/ml, and the cold hemagglutinin titer was high. Pulmonary-function tests showed mixed ventilatory dysfunction, and arterial blood gas analysis revealed a PaO2 of 55.8 Torr on room air. Pseudomonas aeruginosa was cultured from her sputum. A chest X ray film and CT scan showed diffuse nodular shadows and bronchiectatic changes with mild hyperinflation. An infiltrative lesion in right S6 area could also be seen. Administration of broad-spectrum antibiotics did not alleviate her symptoms. The level of myeloperoxidase-specific antineutrophil cytoplasmic antibody (MPO-ANCA) in serum was 245 EU/ml, and 67Ga scintigraphy showed marked accumulation in the abdomen. Abdominal angiography demonstrated a bead-like appearance and irregularities in the peripheral branches of the hapatic artery, the splenic artery, the cystic artery, and the superior mesenteric artery. Because of the high MPO-ANCA level and the angiographic abnormalities, MPO-ANCA related vasculitis was diagnosed. She was treated with 1 g of methylprednisolone daily for 3 days, followed by 60 mg of prednisolone and 50 mg of cyclophosphamide daily. Her condition improved dramatically, and the MPO-ANCA level became almost normal. During treatment, her blood pressure rose markedly with a normal serum creatinine level and normal urinalysis. Plasma renin activity was 13.3 ng/ml/hr. Renal angiography showed stenoses and irregularities in the peripheral branches of renal arteries bilaterally. These findings led to a diagnosis of renovascular hypertension due to vasculitis. Her blood pressure was controlled with an angiotensin-converting enzyme inhibitor and a calcium antagonist. Vasculitis associated with chronic supportive lung disease has occasionally been reported, which suggests a casual relation between chronic respiratory infection and ANCA related vasculitis. Systemic vasculitis should be taken into account as a potential complication of chronic suppurative lung disease. PMID- 9742864 TI - [Idiopathic interstitial pneumonia exacerbated by alveolar hemorrhage]. AB - A 60-year-old man, in who idiopathic interstitial pneumonia (IIP) was diagnosed in 1990, was admitted to our hospital in March 1995 with exertional dyspnea, hemosputum and fever. On chest X ray, new infiltrates in the bilateral middle lung fields were seen, in addition to reticulonodular shadows in the bilateral lower lung fields, which had been noted in 1990. Intubation was reformed because of hypoxemia, and bronchoalveolar lavage (BAL) was performed in right B6. The BAL fluid was bloody, and a diagnosis of alveolar hemorrhage was made based on the presence of many hemosiderin-laden macrophages in the fluid. After pulse therapy with methylprednisolone, the hypoxemia and chest X ray findings improved, and he was extubated. Alveolar hemorrhage may be one possible complication leading to an exacerbation of IIP. PMID- 9742865 TI - [A pulmonary tumorlet with caseous granuloma associated with atypical mycobacterium]. AB - We encountered a case of pulmonary tumorlet with caseous granuloma associated with atypical mycobacterium. A 73-year-old woman was admitted to the hospital because a chest x-ray film showed enlargement of an abnormal shadow in the middle lobe of the right lung. Primary lung cancer was suspected and right middle lobectomy was performed. Acid-fast bacilli (Gaffky 1) were found in a caseous lesion and examination of intraoperatively obtained frozen specimens showed caseous granulomas. The bacilli were later identified as Mycobacterium avium complex. The permanent specimen showed a minute lesion consisting of small clusters of epithelial cells resembling carcinoid tumor in contact with granulomatous tissue. Histopathological examination revealed argyrophilia on Grimelius stain and immunoreactivity to chromogranin-A in the clusters of epithelial cells. Although these results are consistent with small cell carcinoma or peripheral carcinoid tumor, pulmonary tumorlet was diagnosed because of the lesion's small and minimal cytologic atypia, and because of chronic pulmonary damage around the lesion. Pulmonary tumorlets are minute, usually microscopic, tumor-like lesions mostly found in damaged lung tissue obtained at autopsy or during surgery. Morphological diagnosis is sometimes very difficult, but recently these lesions have been regarded as hyperplastic lesions arising in pulmonary neuroendocrine cells (Kultschitzky cells) and caused by chronic pulmonary damage, such as hypoxia and inflammation. Pulmonary tumorlets must be considered in the differential diagnosis of minute lesions suspected to be small cell carcinoma or peripheral carcinoid tumor. PMID- 9742866 TI - [Total collapse of the right lung in a patient with allergic bronchopulmonary aspergillosis]. AB - A 29-year-old man was admitted to the hospital because of a high fever and dyspnea. He had a history of bronchial asthma and had had a bullectomy of the right lung at 15 years of age. He had visited a family physician because of fever and non productive coughing. Medications had no effect on his symptoms, and dyspnea developed. A chest X-ray film showed total collapse of the right lung, and he was referred to our hospital. Laboratory data showed eosinophilia and a high titer of IgE. Total obstruction of the right main stem bronchus by mucous plug was found during fiberoptic bronchoscopy. Aspergillus was detected by pathological examination of bronchial lavage fluid. Tests for aspergillus specific IgE and IgG antibody were positive, as was immediate skin reactivity to Aspergillus. Allergic bronchopulmonary aspergillosis (ABPA) was diagnosed. Infusion and inhalation of a corticosteroid and fluconazole were effective; the symptoms resolved and X-ray findings improved. While migratory infiltration, proximal bronchiectasis and segmental or subsegmental atelectasis caused by a mucous plug are common X-ray findings in allergic bronchopulmonary aspergillosis, total collapse is rare. PMID- 9742867 TI - [Chylothorax resulting from malignant non Hodgkin's lymphoma]. AB - We report a case of chylothorax with malignant non-Hodgkin's lymphoma. A 51-year old man was admitted to our hospital with a chief complaint of dyspnea on March 18, 1995. An x-ray film of the chest revealed a large right pleural effusion. Biochemical tests of the pleural fluid revealed chyle. CT scans of abdomen and chest showed retroperitoneal and mediastinal tumors. Examination of biopsy specimens of the retroperitoneal tumor showed non-Hodgkin's lymphoma (diffuse small cell type, B cell). Treatment with chemotherapy (CHOP, ProMACE-CytaBOM) resulted in complete remission. The chylothorax disappeared after chemotherapy. Only 21 cases of malignant lymphoma with chylothorax have been reported in Japan to date, but their numbers are gradually increasing. We must consider malignant lymphoma in patients with chylothorax. PMID- 9742868 TI - [Primary Sjogren's syndrome with pulmonary hypertension]. AB - A 50-year-old woman was admitted to our hospital because of dyspnea on exertion. A chest X-ray film showed cardiomegaly and echocardiography revealed right-heart overload. Right heart catheterization studies revealed pulmonary hypertension and no signs of pulmonary embolism or vasculitis. The patient had complained of Raynaud's phenomenon since 6 years before admission. She did not notice eye or mouth dryness, but examination of a lip-biopsy specimen showed infiltration of lymphocytes into the salivary glands. Laboratory findings showed hypergammaglobulinemia and hemolytic anemia. Primary Sjogren's syndrome with pulmonary hypertension was diagnosed. Her general condition improved with steroid pulse therapy followed by oral administration of corticosteroids and cyclophosphamide. PMID- 9742869 TI - [Severe bronchorrhea accompanying alveolar cell carcinoma: treatment with clarithromycin and inhaled beclomethasone]. AB - We report the efficacy of oral clarithromycin and inhaled beclomethasone against severe bronchorrhea in a patient with alveolar cell carcinoma. A 54-year-old man produced about 500 to 900 ml of clear and egg-white-like sputum each day. Anti cancer chemotherapy and erythromycin therapy did not reduce the volume of sputum. After administration of clarithromycin and inhaled beclomethasone, sputum volume decreased to about 300 nl each day and the patient's ability to perform daily activities improved. Two months later, clarithromycin was stopped and the patient was treated with inhaled beclomethasone alone. Sputum volume did not increase for 6 months, although the chestroentgenographic findings gradually worsened. Then the sputum volume gradually increased. Five months after the sputum volume began to increase, he was producing about 2 liters of sputum each day and died of respiratory failure. Although the levels of CA 19-9, SLX, and CEA in serum were all within the normal range, the sputum contained high levels of CA 19-9 (1,133,620 U/ml), SLX (3,000 U/ml), and CEA (283 ng/ml). In patients with bronchorrhea, measurement of tumor markers in sputum may be useful for the diagnosis of alveolar cell carcinoma. PMID- 9742870 TI - [Familial pulmonary arteriovenous malformation diagnostic of the Osler-Weber Rendu disease]. AB - A 53-year-old woman was admitted to the hospital for investigation of an abnormal shadow in the middle lobe of the right lung on a chest X-ray film. The filling of most of the shadowed area with contrast media on a chest CT scan suggested a vascular lesion. Further investigation by intravenous digital subtraction angiography and color doppler sonography led to the diagnosis of pulmonary arteriovenous malformation. An abdominal CT scan suggested the presence of an arteriovenous malformation in the liver, and a bronchoscopic study revealed telangiectatic lesions on the laryngeal mucosa. The patient's father had died at the age of 61, due to intracranial bleeding, which suggested he might have had an arteriovenous malformation in the brain. The patient's daughter has a pulmonary arteriovenous malformation. This family of vascular malformation indicates that the patient had Osler-Weber-Rendu disease. Osler-Weber-Rendu disease is an autosomal-dominant inherited disorder that may give rise to arteriovenous malformations, mainly in the lungs and the brain. We found that color doppler sonography can be useful for the diagnosis of pleural-based pulmonary arteriovenous malformation. PMID- 9742871 TI - [Heerfordt's syndrome remitting without corticosteroid therapy]. AB - A 25-year-old woman presented with low-grade fever, uveitis, and bilateral swelling of the parotid glands. Her older sister had a history of sarcoidosis. A chest X-ray film showed bilateral hilar lymphadenopathy. The low-grade fever persisted after admission. Ga scintigraphy showed abnormal uptake in the parotid glands and hilar lymph nodes bilaterally. Sarcoidosis was diagnosed histologically after epithelioid cell granulomas without caseous necrosis were found in a specimen obtained by transbronchial biopsy. Heerfordt's syndrome was the final diagnosis. Six days after admission, left facial and left trigeminal nerve paralysis developed. The symptoms remitted without steroid therapy and, as of eight months after discharge there had been no evidence of recurrence. PMID- 9742872 TI - [Pulmonary pseudallescheriasis in a patient with diabetes mellitus and alcoholic liver cirrhosis]. AB - A 62-year-old man with diabetes mellitus and alcoholic liver cirrhosis was admitted to the hospital because of hemoptysis. Chest X-ray films and computed tomograms showed a dense infiltrative lesion and a healed tuberculous cavity with a possible fungus ball in the upper lobe of the right lung. Bronchoscopy revealed that the hemoptysis originated from the right upper-lobe bronchus. The bleeding stopped after thrombin was applied into the bronchus. Filamentous fungi were seen in lavage fluid from the right upper-lobe bronchus. The fungi were identified as Pseudallescheria boydii, and pulmonary pseudallescheriasis was diagnosed. the patient was treated successfully with miconazole (400 mg/day) for 2 months. Pseudallescheriasis should be taken into account in the differential diagnosis of aspergilloma-like lesions. PMID- 9742873 TI - [Proton magnetic resonance spectroscopy and single photon emission CT in patients with olivopontocerebellar atrophy]. AB - Using proton magnetic resonance spectroscopy (1H-MRS) and single photon emission CT (SPECT), the cerebellum of patients with olivopontocerebellar atrophy (OPCA) and of age-matched control subjects was studied. A spectrum was collected from a 27 cm3 (3 x 3 x 3 cm) voxel in the cerebellum containing white and gray matters in order to measure the distribution and relative signal intensities of N acetylaspartate (NAA), creatine (Cre) and choline (Cho). In the cerebellum of the patients with OPCA, mean NAA/Cre ratios for OPCA patients were significantly decreased compared with normal control subjects (OPCA, 1.01 +/- 0.247; controls, 1.526 +/- 0.144: p < 0.001). Mean NAA/Cho ratios for OPCA patients were slightly decreased (OPCA, 1.285 +/- 0.228; controls 1.702 +/- 0.469: p < 0.06). Cho/Cre ratios valued in the cerebellum of OPCA patients were not significantly different from those in normal controls (OPCA, 0.793 +/- 0.186; controls, 0.946 +/- 0.219). The ratio of RI count in the cerebellum to that in the occipital lobe was significantly decreased in OPCA patients (OPCA, 0.947 +/ 0.096; controls, 1.06 +/ 0.063: p < 0.01). Cerebellar signs were assessed including gait ataxia, limb ataxia, dysarthria, saccadic pursuit, and nystagmus separately or in combination. In patients with more severe ataxic gait and dysarthria. MRS revealed slightly lowered NAA/Cre ratio. There was no significant correlation between NAA/Cre ratio and severity of other clinical signs. The MRS and SPECT findings give a confirmative evidence of hypofunction in cerebellum of patients with OPCA. PMID- 9742874 TI - Problem in prolonged levodopa administration of Parkinson disease patients--from the standpoint of the autonomic nervous function. AB - We studied 50 patients with Parkinson's disease to determine any relation of epidemiologic factors (such as the duration of illness), the duration of levodopa therapy, the total amount of administered levodopa to degree of autonomic dysfunction. Autonomic dysfunction was evaluated using the composite autonomic scoring scale (CASS). The CASS scores were significantly higher among patients with greater administration amount of levodopa regardless of the duration of illness (p < 0.05). There was a definite positive correlation between cardiovascular heart rate index, which was one of the parameters of CASS and reflected parasympathetic neuron system function of cardiac origin, and the duration of levodopa therapy (r = 0.44, p < 0.01) or the total administered amount of levodopa (r = 0.43, p < 0.01). We conclude that long-term levodopa therapy negatively affects autonomic nervous function. PMID- 9742875 TI - [A case of severe hypernatremia complicated with rhabdomyolysis]. AB - We report a 45-year-old male patient with severe hypernatremia followed by rhabdomyolysis and acute renal failure. He had developed hypernatremia for two years after a surgery for an intraventricular AVM involving the hypothalamic area, which was fed by the anterior cerebral artery. He was admitted to our hospital because of progressive muscle weakness. Because of loss of water intake due to impaired thirst sensation, he developed severe hypernatremia (191mEq/l) and marked rhabdomyolysis (CK 17,772 IU/l). He was treated with fluid supplement and hemodialysis for acute renal insufficiency. He had no thirst feeling. Blood studies revealed hyporeactivity to ADH in spite of marked hypernatremia and hyperosmorality. Therefore his condition was considered as adipsic hypernatremia. We concluded that rhabdomyolysis of this case was caused by severe hypernatremia. On reviewing the literature, only a few cases of rhabdomyolysis due to hypernatremia have been reported. PMID- 9742876 TI - [A case of hypothalamic hamartoma manifesting gelastic seizure and multifocal independent seizure foci]. AB - A 25-year-old man manifesting gelastic seizure in association with hypothalamic hamartoma was reported. The epileptogenic focus was evaluated by long-term video EEG monitoring, single photon emission computed tomography (SPECT) and positron emission tomography (PET). Multiple independent ictal foci were found in the ictal EEG recording, and glucose metabolism and cerebral blood flow were decreased in scattered cortical areas. Facial asymmetry seen during gelastic seizure in this case may be caused by contraction of mimetic contralateral to the ictal focus in the temporal lobe. PMID- 9742877 TI - [An adult case of Reye syndrome induced by diclofenac sodium, and recovered by plasma exchange]. AB - Reye syndrome (RS) is an acute encephalopathy in childhood, and is very rare in adulthood. Here we report a 21-year-old woman with RS. Because of her dysmenorrhea, she took 3 tablets of diclofenac sodium (25 mg) per day in 3 divided doses for two days. Two days after the last intake of the medicine, she developed high fever, nausea, vomiting, and disturbance of consciousness with delirium, i.e., acute encephalopathy. She did not have seizure, hemiplegia, or other focal neurological manifestations. The serum GOT level was normal at onset, but in 12 hours dramatically increased up to 8,632 IU/L. The serum bilirubin level was normal. The cerebrospinal fluid revealed normal cell count, and protein. Although the liver biopsy was not performed because of thrombocytopenia, we diagnosed her as an adult case of RS according to the clinical criteria of the Center for Disease Control. In addition to treatment for the brain edema, plasma exchange was performed once treat the encephalopathy at the onset. The next day, her consciousness level and serum GOT level markedly improved. She completely recovered from acute encephalopathy in a week after her admission. In conclusion, diclofenac sodium, as well as aspirin, should be considered as a possible causal agent for RS, and early plasma exchange may be beneficial. PMID- 9742878 TI - [Chronic Cryptococcal meningitis with CSF oligoclonal IgG band in a patient with Claude syndrome]. AB - We described a 61-year-old man who was diagnosed as having chronic cryptococcal meningitis, while he was hospitalized with Claude syndrome. The patient was admitted because of acute onset of gait disturbance. He had a tendency to fall down to his left side since he awoke in the morning of August 12, 1995. On admission, he was mentally alert, showing a right oculomotor nerve palsy, gaze evoked horizontal nystagmus in the left eye on the left lateral gaze, and incoordination of the left upper and lower extremities. In addition, he fell to the left side on standing up with feet together and with eyes closed. He had mild wild-based gait with a tendency to fall down to the left on tandem gait. Babinski sign was present on the left side. He did not have fever, nor meningeal signs, nor sensory abnormalities. X-ray films of the chest showed multiple nodular shadows consistent with pneumoconiosis. Cranial X-ray computed tomography and magnetic resonance imaging revealed a small lesion in the paramedian area of the midbrain on the right, consistent with an infarct. Cerebral arteriography revealed a stenosis in the proximal portion of the right posterior cerebral artery. Cerebrospinal fluid (CSF) showed a moderate mononuclear cell predominant pleocytosis, a moderate elevation of total protein, slightly reduced glucose values. Although the culture and India ink preparation of CSF were negative for cryptococcus in repeated studies, its antigen was positive both in the serum and CSF. In addition, the CSF showed an oligoclonal IgG band which was predominantly K type. After the antigen of Cryptococcus neoformans was added to the CSF in vitro, the oligoclonal IgG band was absorbed completely. The patient was treated with fluconazole (FLCZ), which did not cause any improvement of the CSF abnormalities, so that FLCZ was replaced by 5-flucytosine (5-FC). Since the CSF abnormalities moderately improved with 5-FC, he was discharged on December 21, 1995. After the 5-FC was discontinued, the CSF results slowly worsened over several months without any signs and symptoms of meningitis. He was hospitalized again on October 28, 1996 for treatment with both 5-FC and amphotericin B. Although the CSF abnormalities improved markedly, the meningitis was not cured. After he was discharged on February 1, 1997, he was treated with both 5-FC and FLCZ. Although his CSF abnormalities worsened mildly, he remained afebrile without meningeal signs and symptoms and led an ordinary life. In our patient it remained undetermined whether the Claude syndrome was caused by arteriosclerotic infarction, or vasculitis due to cryptococcal meningitis, or both. Asymptomatic chronic cryptococcal meningitis as observed in our patients is unusual. In addition, this is the second case after Porter et al (1977) that the oligoclonal IgG band in CSF proved to be related to cryptococcal infection. PMID- 9742879 TI - [Bickerstaff's brainstem encephalitis associated with nystagmus]. AB - A 29-year-old man developed drowsiness, ophthalmoplegia and cerebellar ataxia following upper respiratory tract infection. We diagnosed the patient as having a Bickerstaff's brainstem encephalitis. There was upbeating nystagmus which appeared with upward gaze, and bilateral horizontal gaze-evoked nystagmus in both directions. On electronystagmography, eye-tracking test showed saccadic pattern with impaired smooth pursuit eye movement. The horizontal optokinetic nystagmus test showed diminution of response. These results suggested that brainstem and cerebellum were widely involved. Presence of nystagmus may help to speculate the lesion of Bickerstaff's brainstem encephalitis. PMID- 9742880 TI - [Giant cell arteritis with bilateral obstruction of the internal carotid artery- report of an autopsy case]. AB - Internal carotid artery involvement and dementia occur infrequently in patients with giant cell (temporal) arteritis. A 75-year-old woman admitted with progressive cognitive decline, drowsiness and headache was diagnosed as having giant cell arteritis by temporal artery biopsy (TAB). High dose corticosteroid improved inflammatory reaction but did not improve his cognitive function. Cerebral angiograms showed obstruction of both internal carotid arteries at the siphon. Brain MRI showed only small cerebral infarcts in the basal ganglia and corona radiata bilaterally. However, brain SPECT disclosed reduced cerebral blood flow in the frontal lobe bilaterally. A postmortem examination revealed bilateral parietal infarcts and isolated giant cell arteritis involving the both internal carotid arteries at the siphon. We speculated that perfusion insufficiency and multiple cerebral infarction due to bilateral internal carotid artery occlusion had caused this neurologic deterioration. PMID- 9742881 TI - [Disappearance of embolic signals on transcranial Doppler sonography following antiplatelet therapy in a patient with transient ischemic attacks]. AB - Transcranial Doppler sonography (TCD) has been increasingly used to detect embolic signals. A 49-year-old woman had repeated episodes of weakness and numbness of the right limbs. She was diagnosed as having transient ischemic attacks (TIAs). A cranial Gadolinium-enhanced MRI revealed multiple small lesions in the cortical-subcortical areas of the left middle cerebral artery territory. A TCD examination on admission detected 4 embolic signals on the left MCA trunk in a 30-minute examination period. TIAs completely ceased after the initiation of an antiplatelet therapy with aspirin. Embolic signals were not detected in later TCD studies. The embolic signals on the TCD strongly indicated that TIAs are caused by an embolic mechanism. TCD is potentially useful for assessing the efficacy of antiplatelet and anticoagulant therapies in patients with TIAs. PMID- 9742882 TI - [A case of pharyngeal-cervical-branchial variant of Guillain-Barre syndrome with IgG anti-GT1a antibody]. AB - A 36-year-old man with pharyngeal-cervical-brachial variant of Guillain-Barre syndrome (PCB) was described. Neurologic examination revealed total ophthalmoplegia, pharyngeal-cervical-brachial weakness and hyporeflexia in the upper limbs, sparing power and tendon reflexes in the lower limbs. Enzyme-linked immunosorbent assay showed that he had high titer of IgG antibody to GT1a (1:32,000), which did not cross-react with GQ1 b or GD1a. Thin-layer chromatography immunostaining confirmed that his serum IgG reacted with GT1a. These findings show that IgG anti-GT1a antibody without cross-reactivity with GQ1b plays a role in the development of PCB. PMID- 9742883 TI - [Hypouricemia in patients with meningitis]. AB - Serum urate and sodium concentrations were measured in 23 patients with acute viral and bacterial meningitis. Serum urate level was 3.0 +/- 0.2 mg/dl (mean +/- S.D.) (3.6 +/- 1.2 mg/dl in male and 2.5 +/- 0.9 mg /dl in female) on admission, but gradually elevated with improvements of meningitis. It turned to 4.8 +/- 0.2 mg/dl after recovery, and the value on admission was significantly lower than that after recovery (p < 0.0001). Serum sodium level was 137.6 +/- 2.9 mEq/l on admission and 139.7 +/- 2.7 mEq/l after recovery; also lower in the former (p < 0.01). These results show that patients develop transient hypouricemia, which may be explained by SIADH (syndrome of inappropriate secretion of ADH), although SIADH is subclinical in most cases of meningitis. PMID- 9742884 TI - [A case of paradoxical brain embolism in a 17-year-old boy]. AB - A 17-year-old healthy high school boy suddenly suffered from consciousness disturbance immediately after a karate match. His neurological findings on admission were consciousness disturbance, motor aphasia, right hemiparesis, and right Babinski reflex. Cerebral angiography showed occlusion of the precentral branch of the left middle cerebral artery, and X-ray CT scan on the second day revealed low density area in its territory. No abnormality was found on transthoracic echocardiography or Holter ECG. Contrast transesophageal echocardiography with Valsalva maneuver was performed twice (the 2nd and the 14th day), and patent foramen ovale (PFO) was found only on the second trial. It was supposed that paradoxical embolism had occurred. Transesophageal echocardiography is necessary for the embolic patients of unknown origin, especially young adults. To find PFO, it is important to repeat contrast transesophageal echocardiography carefully. if it is negative on the first trial. PMID- 9742885 TI - [Unilateral gustatory disturbance by pontine infarction]. AB - We reported a case of pontine infarction presenting as diminished taste on the contralateral side. A 67-year-old man was hospitalized with a sudden onset of right hypogeusia. No neurological abnormalities were found except diminished taste in the areas innervated by the chorda tympani, greater petrosal and glossopharyngeal nerves on the right side. Brain MRI demonstrated a lesion with low-intensity on T1-weighted images and high-intensity on T2-weighted images in the left suprapontine tegmentum. Cases of pontine disease presenting as contralateral dysgeusia have rarely been reported. In the present case, we considered that infarction occurred superior to the pontine taste are (PTA). It is suggested that the gustatory pathway superior to PTA takes a chiasmal tract ascending in the brainstem. PMID- 9742887 TI - [Molecular mechanisms of fat-soluble vitamins: vitamin receptor knockout mice]. PMID- 9742886 TI - [Persistent lateral gaze palsy and abducens nerve palsy due to pontine infarction]. AB - A 76-year-old hypertensive man noticed sudden dizziness and diplopia. On examination, right-sided gaze deviation was present, but the remainder of the neurological examination was normal. Five days later, MRI revealed a small lesion in the left paramedian pontine tegmentum, which was consistent with pontine infarction. The conjugate deviation disappeared about two weeks after the onset, but a left lateral gaze and abducens nerve palsy have persisted at ten months after the infarct. MRI findings are unchanged. The site of infarction is presumed to be located in the caudal portion of paramedian reticular formation and the abducens nerve fascicle. Persistent lateral gaze palsy is rare in pontine infarction, and this case is important in illustrating the anatomical location of paramedian pontine reticular formation. PMID- 9742888 TI - [The essential character of the Spemann's organizer]. PMID- 9742889 TI - [Brachyury genes and notochord differentiation in chordate embryos]. PMID- 9742890 TI - [A functional role for PDZ proteins in synapse organization and synaptic plasticity]. PMID- 9742891 TI - [Single molecule imaging of biological functions]. PMID- 9742892 TI - [Making of guideline for gene diagnosis in familial tumors]. PMID- 9742893 TI - Bladder cancer: twenty years of progress and the challenges that remain. AB - The most obvious improvements in the care of patients with bladder cancer over the past 20 years are bacille Calmette-Guerin immunotherapy for superficial bladder cancer and cisplatin and methotrexate-based combination chemotherapy for advanced disease. The challenges that remain are prevention, early detection, and improved treatment of both superficial and advanced disease. Meeting these challenges requires research, financial investment, and public education. PMID- 9742894 TI - Bladder cancer: state-of-the-art care. AB - The challenge in bladder cancer is to control superficial disease and prevent its recurrence or progression. Patients with invasive disease need to be identified earlier, when disease may be less advanced and more amenable to cure. An important area for further investigation is the biology of the various forms of bladder cancer and the various pathways of development they may follow. PMID- 9742895 TI - Perspective: National Cancer Institute summary report about estimated exposures and thyroid doses received from iodine 131 in fallout after Nevada atmospheric nuclear bomb tests. AB - Environmental 131I contamination from atmospheric nuclear bomb tests conducted at the NTS from 1951 to 1958 exposed Americans nationwide to a cumulative average dose of 1 to 4 rad to the thyroid gland. By comparison, 10 years of exposure to natural background sources of thyroid radiation results in a cumulative dose of 1 rad. Americans living in certain high-deposition areas received an average cumulative thyroid dose of as much as 16 rad. Individual dose rates vary considerably as a function of age at the time of exposure, site of residence, and dietary habits with respect to milk consumption. The individual cumulative thyroid dose for persons born between 1945 and 1958 may be significantly higher than the reported averages for their locale. The NCI report contains voluminous data tables permitting detailed calculations of individual dose. Additionally, color-coded dose maps allow one to approximate individual dose conveniently. Translation of cumulative thyroid dose attributable to 131I to predictions of increased rates of thyroid cancer appears problematic and is the subject of further study. In contrast to studies of patients receiving external thyroid irradiation, existing studies of patients treated with 131I for diagnostic and therapeutic medical purposes do not document increased rates of thyroid cancer. An Institute of Medicine task force is expected to issue a report on this subject in September 1998. This review also briefly summarizes the evaluation, diagnosis, and treatment of patients with papillary and follicular thyroid cancers. Data from 53,856 patients with thyroid cancer accessioned to the NCDB from 1985 to 1995 document extremely high survival rates for patients in the United States with papillary and follicular thyroid cancer. PMID- 9742896 TI - Contemporary issues in the management of endometrial cancer. AB - Several important issues exist in the management of endometrial cancer, including the possible induction of endometrial cancer by tamoxifen: the newly developed surgical staging system; the increasing use of minimal-access surgery, including laparoscopy; the role of postoperative adjuvant therapy; the frequency and extent of follow-up visits and surveillance; and the use of estrogen replacement therapy in women with a history of endometrial cancer. PMID- 9742898 TI - Restoring a physician-led healthcare system. PMID- 9742897 TI - Status report on prostate cancer in African Americans: a national blueprint for action. AB - "A leadership Conference--Prostate Cancer in the African-AMerican Community: an Agenda for Action" was held November 20 to 22, 1997, in Houston, Texas. The meeting--a collaborative effort of the American Cancer Society, The Centers for Disease Control and Prevention, and the National Cancer Institute--examined the unique challenge that prostate cancer poses to the African-American community. This article presents the National Blueprint for Action developed at this historic conference. PMID- 9742899 TI - Do caregivers still care? PMID- 9742900 TI - Risks for renal involvement in diabetes. PMID- 9742901 TI - Relief from perioral dermatitis. PMID- 9742902 TI - A baffling case of bulging belly. Protease paunch. PMID- 9742903 TI - Which culprit is causing your patient's otorrhea? AB - Otorrhea varies in appearance and can be accompanied by a multitude of symptoms. Recognition of what various combinations indicate--whether otitis externa, chronic otitis media, Langerhans' cell histiocytosis, or basal or squamous cell carcinoma--is essential. Physicians must also be familiar with the tests appropriate for confirming otorrhea's cause. Once proper diagnosis is made, effective treatment can follow. PMID- 9742904 TI - Is this lump in the neck anything to worry about? . AB - Neck masses are common and often the result of self-limited inflammation or infection. Congenital masses and neoplasms are additional benign causes. However, a neck mass can signal a malignant tumor, so it is important to understand what represents an abnormal mass and to recognize warning signs and symptoms. Patients at risk of serious clinical problems may be identified through findings on history taking (e.g., progressive enlargement of the mass, prolonged hoarseness) and physical examination (e.g., unilateral nasal obstruction, intraoral induration). Suspicion of a malignant process calls for referral to a subspecialist in head and neck disorders. PMID- 9742905 TI - Hearing loss: the invisible disability. AB - Loss of hearing is a national health problem with significant physical and psychological repercussions. Although there is no cure for certain forms of hearing loss, many patients can be helped, especially when the problem is recognized early. The authors discuss the important role of primary care physicians in early identification, management, and counseling of hearing impaired patients. PMID- 9742906 TI - What role for antibiotics in otitis media and sinusitis? AB - In patients with otitis media or sinusitis, antibiotics must be used judiciously. First-line treatment for both uncomplicated acute otitis media and acute sinusitis is amoxicillin. Erythromycin ethylsuccinate and sulfisoxazole or TMP SMZ may be used in patients who are allergic to penicillin. Beta-lactamase-stable agents should be given when no response occurs within 48 to 72 hours. In cases in which penicillin-resistant pneumococcus is suspected, high-dose amoxicillin, with or without clavulanate, or clindamycin should be considered. Antibiotics are not indicated for initial treatment of otitis media with effusion but may be considered for effusions lasting longer than 3 months. Prophylactic antibiotics should be considered only for recurrent acute infections occurring three or more times within 6 months or four or more times within a year. The common cold should not be treated with antibiotics, and antimicrobial therapy should be initiated only when there is reasonable clinical certainty about the presence of acute sinusitis. PMID- 9742907 TI - When to suspect lactose intolerance. Symptomatic, ethnic, and laboratory clues. AB - Lactose intolerance is widespread, with adult-type hypolactasia being the predominant cause of lactose malabsorption. Daily ingestion of less than 240 mL of milk is well tolerated by most lactose-intolerant adults. Some persons with normal lactase activity may become symptomatic on consumption of products containing lactose. Lactose maldigestion may coexist in adults with irritable bowel syndrome and in children with recurrent abdominal pain. Management consists primarily of dietary changes. People who avoid dairy products should receive calcium supplementation and should be advised to read ingredient labels carefully. Several lactase replacement products are available, but their efficacy varies. PMID- 9742908 TI - Anti-inflammatory drugs for controlling asthma. AB - Anti-inflammatory agents are the first-line treatment for controlling mild persistent, moderate persistent, and severe persistent asthma. The choice of drug and dosage must be individualized to the patient. In general, the glucocorticoids are widely accepted as the most potent and preferred asthma treatment in most adults and some children. Cromolyn, because of its safety and availability in a nebulized form, is the first-line treatment in most young children. The leukotriene inhibitors appear to be effective in mild asthma, but further clinical studies are needed to determine their role more precisely. As the mechanisms of inflammation in asthma are further defined, new pharmaceutical products will be developed to aid in arresting this process. PMID- 9742909 TI - Depression and aging too often do mix. AB - Depression in elderly persons is prevalent, often undiagnosed, and usually untreated. Because there is no reliable diagnostic test, careful clinical evaluation is essential. With aggressive antidepressant treatment, the overall long-term outcome in elderly patients is optimistic. In this article, Dr Birrer discusses the scope of the problem, important diagnostic considerations, and the current recommended treatments in this population. PMID- 9742910 TI - Alpha 1-blocker combination therapy for hypertension. AB - Combination therapy is a cost-effective and rational approach to treatment of severe hypertension and of mild to moderate hypertension that is refractory to monotherapy. The method has several advantages, most notably improved tolerability and enhanced antihypertensive efficacy. Long-term prospective studies are needed to confirm that such agents as calcium channel blockers, ACE inhibitors, and alpha 1 blockers reduce end-organ damage more effectively than do older antihypertensive drugs. However, scientific evidence strongly suggests that reducing risk factors for end-organ damage reduces heart, brain, kidney, and large-artery injury. Alpha 1 blockers appear to be a particularly suitable choice for use in combination regimens. The only class of agents that should be avoided in combination with alpha 1 blockers is central alpha agonists; all other agents act in an additive or synergistic fashion. Unlike diuretics and beta blockers, alpha 1 blockers do not adversely affect serum lipid, glucose, or insulin levels. In fact, alpha 1 blockers may improve these measurements and also counteract the adverse effects of other antihypertensive agents on them. Alpha1-blocker therapy may bring about regression of LVH, and it does not have deleterious effects on disorders that often coexist with hypertension (e.g., gout, chronic obstructive lung disease, peripheral ischemia). PMID- 9742911 TI - The American Surgical Association: past, present, and future. PMID- 9742912 TI - Experience with 500 simultaneous pancreas-kidney transplants. AB - METHODS: From December 1985 to October 1997, 500 simultaneous pancreas-kidney transplants (SPKs) were performed at the University of Wisconsin. Bladder drainage (BD) was used in 388 and enteric drainage (ED) in 112. All pancreas transplants were preserved in UW solution. RESULTS: Patient survival at 1, 5, and 10 years was 96.4%, 88.6%, and 76.3%; kidney function, 88.6%, 80.3%, and 66.6%; and pancreas function, 87.5%, 78.1%, and 67.2%. Thrombosis of the pancreas occurred in three to four (0.6% to 0.8%) and primary nonfunction in one (0.2%). There was a 4.2% acute tubular necrosis rate for the kidney. Conversion from BD to ED was required in 24% of cases. Primary indications for enteric conversion (EC) were leak (14%), urethritis and extravasation (7%), and chronic hematuria (3%). No graft was lost as a result of EC. There was no difference in 1-year graft survival between ED and BD. Leading causes of pancreas loss were rejection in 45 patients and death with a functioning graft in 27 patients. Since June 1995, mycophenolate mofetil was used for immunosuppression (n = 109). One-year survival rates with mycophenolate mofetil are patient, 98.1 %; kidney, 94.2%; and pancreas, 93.1%. Steroid-resistant rejections decreased from 48% to 15%. CONCLUSIONS: This series represents the world's largest experience with SPK, including the longest follow-up for BD pancreatic transplants. Ten-year graft survival rates exceed those of all other transplants, with the exception of HLA identical living-related grafts. This series confirms that SPK is a highly successful procedure for selected diabetic patients with renal failure. PMID- 9742913 TI - Cost-effectiveness of coronary artery bypass surgery in octogenarians. AB - OBJECTIVE: The objective of this retrospective cohort study was to determine whether coronary artery bypass graft (CABG) surgery is effective and cost effective relative to medical management of coronary artery disease (CAD) in the elderly. SUMMARY BACKGROUND DATA: The aging of the U.S population and the improvements in surgical techniques have resulted in increasing numbers of elderly patients who undergo this surgery. The three randomized, controlled trials (RCTs) that established the efficacy of CABG surgery completed patient enrollment from 19 to 24 years ago excluded patients older than 65 years. Although information regarding outcomes of CABG in this population is mainly available in case series, a major lacuna exists with respect to information on quality of life and cost effectiveness of surgery as compared with medical management. METHODS: The authors retrospectively formed surgical and medically managed cohorts of octogenarians with significant multivessel CAD. More than 600 medical records of patients older than 80 years who underwent angiography at our institution were reviewed to identify 48 patients who were considered reasonable surgical candidates but had not undergone surgery. This cohort was compared with 176 patients who underwent surgery. RESULTS: The cost per quality-adjusted life year saved was $10,424. At 3 years, survival in the surgical group was 80% as compared with 64% in the entire medical cohort and 50% in a smaller subset of the medical cohort. Quality of life in patients who underwent surgery was measurably better than that of the medical cohort with utility index scores, as measured by the EuroQoL, (a seven-item quality of life questionnaire) of 0.84, 0.61, and 0.74, respectively. CONCLUSIONS: Performing CABG surgery in octogenarians is highly cost-effective. The quality of life of the elderly who elect to undergo CABG surgery is greater than that of their cohorts and equal to that of an average 55-year-old person in the general population. PMID- 9742914 TI - Durability of complete responses in patients with metastatic cancer treated with high-dose interleukin-2: identification of the antigens mediating response. AB - OBJECTIVE: To determine the durability of complete responses in patients with metastatic melanoma or renal cancer treated with high-dose bolus interleukin-2 (IL-2) as well as the factors associated with the development of a complete response and the antigens mediating clinical responses. METHODS: A consecutive series of 409 patients with either metastatic melanoma or renal cancer who were treated with high-dose bolus IL-2 in the Surgery Branch, National Cancer Institute, between September 1985 and November 1996 have been analyzed with a median potential follow-up of 7.1 years. All patients were treated with 720,000 IU/kg administered by 15-minute intravenous infusions every 8 hours for up to 5 days as clinically tolerated per cycle. Two cycles constituted a treatment course. Tumor-infiltrating lymphocytes (TIL) from melanoma patients were used to clone the genes encoding the tumor antigens responsible for clinical responsiveness. RESULTS: Thirty-three of 409 (8.1%) patients treated with high dose bolus IL-2 achieved a complete response and 37 (9%) achieved a partial response. Complete regression was seen in 6.6% and 9.3% of patients with metastatic melanoma and renal cancer, respectively. Twenty-seven of these 33 completely responding patients (82%) remain in ongoing continuous complete response from 39 to more than 148 months from the onset of treatment. Tumor regressions were seen at virtually all organ sites. The absence of prior treatment with immunotherapy, the total dose of IL-2 administered, and the maximal rebound lymphocytosis after cessation of IL-2 correlated with achieving a complete response. Expression cloning techniques have identified a series of tumor antigens that are recognized by TIL grown from resected melanomas. These antigens are mainly melanoma/ melanocyte differentiation antigens, although mutated intracellular proteins can also serve as antigens. CONCLUSIONS: Treatment with high-dose bolus IL-2 mediates complete cancer regression in approximately 8% of patients with metastatic renal cancer and melanoma. The great majority of these patients will enter durable complete regressions and appear to be cured of their metastatic cancer. Thus, immunotherapy with high-dose bolus IL-2 should be considered as initial therapy for appropriately selected patients with metastatic melanoma and renal cell cancer. Identification of the tumor antigens mediating clinical response is opening new therapeutic possibilities for cancer treatment. PMID- 9742915 TI - The importance of surgeon experience for clinical and economic outcomes from thyroidectomy. AB - OBJECTIVE: To determine whether individual surgeon experience is associated with improved short-term clinical and economic outcomes for patients with benign and malignant thyroid disease who underwent thyroid procedures in Maryland between 1991 and 1996. SUMMARY BACKGROUND DATA: There is a prevailing belief that surgeon experience affects patient outcomes in endocrine surgery, but there is a paucity of objective evidence outside of clinical series published by experienced surgeons that supports this view. METHODS: A cross-sectional analysis of all patients who underwent thyroidectomy in Maryland between 1991 and 1996 was conducted using a computerized statewide hospital discharge data base. Surgeons were categorized by volume of thyroidectomies over the 6-year study period: A (1 to 9 cases), B (10 to 29 cases), C (30 to 100 cases), and D (>100 cases). Multivariate regression was used to assess the relation between surgeon caseload and in-hospital complications, length of stay, and total hospital charges, adjusting for case mix and hospital volume. RESULTS: The highest-volume surgeons (group D) performed the greatest proportion of total thyroidectomies among the 5860 discharges, and they were more likely to operate on patients with cancer. After adjusting for case mix and hospital volume, highest-volume surgeons had the shortest length of stay (1.4 days vs. 1.7 days for groups B and C and 1.9 days for group A) and the lowest complication rate (5.1 % vs. 6.1% for groups B and C and 8.6% for group A). Length of stay and complications were more determined by surgeon experience than hospital volume, which had no consistent association with outcomes. CONCLUSIONS: Individual surgeon experience is significantly associated with complication rates and length of stay for thyroidectomy. PMID- 9742916 TI - Cloaca, the most severe degree of imperforate anus: experience with 195 cases. AB - OBJECTIVE: To provide a follow-up of 195 patients with cloacal malformations seen by the author from 1959 to 1998. SUMMARY BACKGROUND DATA: Cloaca, which occurs in approximately 1 of 50,000 births, is the most complex type of imperforate anus with confluence of the rectum, vagina, and bladder in a urogenital sinus. Functional results for the bowel, the genital tract, and the urinary tract were formerly poor. Cloacal exstrophy, which is an even more complex spectrum of malformations, was uniformly fatal until 1960. In addition to imperforate anus, these babies have an omphalocele, two exstrophic bladders, between which there is an open cecum, and a blindly ending colon hanging down in the pelvis from the cecum. Although both of these diagnoses contain the word "cloaca," which is Latin for sewer, they are really two separate entities in terms of surgical management. Cloaca and cloacal exstrophy in most cases are very different anatomic problems. However, there are variants that are like a hybrid, which is the rationale for reporting together an experience with both entities. METHODS: Records were reviewed of 154 patients with cloaca and 41 patients with cloacal exstrophy to assess anorectal function, urinary continence, and sexual function where available. RESULTS: Follow-up was available in 141 cloaca patients: 82 have spontaneous bowel movements and satisfactory control, 38 use enemas to evacuate, 9 have a colostomy, 7 have fecal soiling, and 5 are too recently operated to evaluate. Regarding urinary control, 83 void spontaneously, 40 catheterize to empty, 4 have urinary diversion, 1 has a continent diversion, 5 patients are wet, and 8 are too recently operated to judge. Twenty-four patients are now adults, 17 of who have experienced coitus and 7 have not. Seven have had babies, all except one by cesarean section. Results of surgery for cloacal exstrophy are not as good, but are encouraging nonetheless for an anomaly that was uniformly fatal before 1960. Of the 41 cloacal exstrophy patients being followed, 7 have not undergone surgery. Fifteen have a colostomy; 19 had pull-through of the colon, but 3 were subsequently reversed for fecal incontinence. Most depend on enemas to evacuate. Urinary dryness was attained in 30 patients, usually by intermittent catheterization of the bladder, which was augmented with small bowel or stomach or both. Only three void voluntarily. Fifteen of the completed long-term patients wear no bag. Only three of the completed patients wear two bags. The rest have one bag. CONCLUSIONS: Imperforate anus and associated malformations in cloaca and cloacal exstrophy are not hopeless problems. A reasonable lifestyle can be achieved for most of these children with comprehensive surgical planning. PMID- 9742917 TI - Prospective randomized trial of early postoperative intraperitoneal chemotherapy as an adjuvant to resectable gastric cancer. AB - OBJECTIVE: Surgeons have postulated on numerous occasions that cancer resection may participate in the dissemination of a malignancy. This randomized trial sought to determine whether a large volume of chemotherapy solution used perioperatively to flood the peritoneal cavity could eliminate microscopic residual disease and thereby improve survival of patients with gastric cancer. SUMMARY BACKGROUND DATA: Surgical treatment failures in patients with gastric cancer are confined to the abdomen in most patients. Resection site and peritoneal surface spread, along with liver metastases, are the most common areas of recurrence. Survival and quality of life of patients with gastric cancer would be improved if disease progression at these anatomic sites was reduced. METHODS: In a prospective randomized trial of 248 patients, intraperitoneal mitomycin C on day 1 and intraperitoneal 5-fluorouracil on days 2 through 5 were administered after gastric cancer resection. Patients who were thought to have stage II or stage III disease were randomized after resection to surgery alone versus surgery plus early postoperative intraperitoneal chemotherapy. After final pathologic examinations, there were 39 patients with stage I, 50 with stage II 95 with stage III, and 64 with resected stage IV cancer. RESULTS: The 5-year survival of the surgery-only group was 29.3%, and the surgery-plus-intraperitoneal chemotherapy group was 38.7% (p = 0.219). In a subset analysis, the patients with stage I, stage II, and stage IV disease showed no statistically significant difference in survival. The 5-year survival rate of patients with stage III disease who underwent surgery only was 18.4% versus a survival rate of 49.1% for patients who underwent surgery plus intraperitoneal chemotherapy (p = 0.011). CONCLUSIONS: In a subset analysis, patients with stage III gastric cancer have shown a statistically significant improvement in survival when treated with perioperative intraperitoneal chemotherapy. Further studies in patients with gastric cancer with surgically directed chemotherapy are suggested. PMID- 9742919 TI - Cancer gene therapy using a replication-competent herpes simplex virus type 1 vector. AB - OBJECTIVE: The authors investigate the efficacy of hrR3, a viral vector derived from herpes simplex virus type 1 (HSV 1), in destroying colon carcinoma cells in vitro and in vivo. The effect of adding the prodrug ganciclovir in combination with hrR3 infection also is assessed. SUMMARY BACKGROUND DATA: Most cancer gene therapy strategies use viral vectors that are incapable of replication. The HSV 1 vector hrR3 is capable of replication, and its replication is cytotoxic to cells. hrR3 also possesses the HSV-thymidine kinase gene, which converts ganciclovir into a toxic metabolite. Thus, the addition of ganciclovir to hrR3-infected cells may enhance the ability of hrR3 to destroy tumor cells. To increase specificity for tumor cells, hrR3 has a mutated ribonucleotide reductase gene and replicates selectively in cells with high levels of endogenous rbonucleotide reductase. Actively dividing cells such as tumor cells have high levels of endogenous ribonucleotide reductase for synthesis of DNA precursors. The authors are interested in the use of HSV 1 vectors to treat liver metastases from colorectal cancer. METHODS: Ribonucleotide reductase expression in several colon carcinoma cell lines and in primary cultures of human hepatocytes was determined by Western blot analysis. hrR3-mediated cytotoxicity in the colon carcinoma cell lines was determined using an in vitro assay. The human colon carcinoma cell line HT29 was injected into the flanks of nude mice followed by intratumoral injection of hrR3. Tumor growth rate was assessed with and without the addition of intraperitoneal ganciclovir. RESULTS: Ribonucleotide reductase levels in colon carcinoma cell lines are much higher than in primary cultures of human hepatocytes. hrR3 efficiently destroys colon carcinoma cell lines in vitro. A single intratumoral injection of hrR3 into HT29 flank tumors significantly reduces tumor growth rate, and the administration of ganciclovir has no additive effect. CONCLUSIONS: The inherent cytotoxicity of hrR3 replication effectively destroys colon carcinoma cells in vitro and in vivo. This cytotoxicity is not enhanced in vivo by the addition of ganciclovir. In the future, more efficacious and selective HSV 1 vectors may be useful in the treatment of cancer. PMID- 9742918 TI - Retroperitoneal soft-tissue sarcoma: analysis of 500 patients treated and followed at a single institution. AB - OBJECTIVE: To analyze treatment and survival of a large cohort of patients with retroperitoneal soft-tissue sarcomas (STS) treated and prospectively followed at a single institution. SUMMARY BACKGROUND DATA: Retroperitoneal STS are relatively uncommon and constitute a difficult management problem. Although surgical resection is often difficult or impossible, current chemotherapy is not effective and radiation is limited by toxicity to adjacent structures. Thus, complete surgical resection remains the most effective modality for selected primary and recurrent disease. METHODS: Five hundred patients with retroperitoneal STS were admitted and treated between July 1, 1982, and September 30, 1997, and prospectively followed. Patient, tumor, and treatment variables were analyzed for disease-specific and disease-free survival. Survival was determined with the Kaplan-Meier method. Statistical significance was evaluated using the logrank test for univariate influence and Cox model stepwise regression for multivariate influence. RESULTS: Two hundred seventy-eight patients (56%) had primary disease and 222 (44%) recurrent disease. Median follow-up was 28 months (range 1 to 172 months), 40 months for survivors. Median survival was 72 months for patients with primary disease, 28 months for those with local recurrence, and 10 months for those with metastasis. For patients with primary or locally recurrent tumors, unresectable disease, incomplete resection, and high-grade tumors significantly reduced survival time. CONCLUSIONS: In this study of patients with retroperitoneal STS, stage at presentation, high histologic grade, unresectable primary tumor, and positive gross margin are strongly associated with the tumor mortality rate. Patients approached with curative intent should undergo aggressive attempts at complete surgical resection. Incomplete resection should be undertaken only for symptom relief. PMID- 9742920 TI - Overestimation of hereditary breast cancer risk. AB - OBJECTIVE: To find out how women with breast or ovarian cancer rate their chances of carrying hereditary factors for these cancers and to determine the extent to which they overestimate their risk. SUMMARY BACKGROUND DATA: BRCA1 and BRCA2 are genes that cause breast and ovarian cancer when they are inherited in families. Testing for disease-associated mutations in these genes is now available commercially. Previous studies have shown that women overestimate their chances of carrying mutations. However, women's perceptions of risk have not been compared to objective estimates or to actual BRCA1 and BRCA2 testing results. METHODS: This study examines estimates of carrying BRCA1 and BRCA2 mutations among women participating in a randomized trial comparing alternative precounseling educational materials. Estimates were provided by participants in a baseline mailed survey. Estimates given by participants were compared to those given by an expert panel and by a statistical model. Testing was offered free of charge and was done in an academic laboratory using standard techniques. Baseline estimates of participating women were compared to the estimates of the expert panel, to the carrier probability provided by the statistical model, and to actual testing results. RESULTS: Women who have a personal history of breast or ovarian cancer significantly overestimate their risk of carrying hereditary factors for breast and ovarian cancer. Self-estimates exceeded the estimates of experts and a statistical model. One hundred women completed testing, and 21 mutations in BRCA1 or BRCA2 were found. Many test-negative women also overestimated their hereditary risk. Some women with a high carrier probability were negative for BRCA1 and BRCA2 mutations. CONCLUSIONS: Overestimation of hereditary factors is common among affected women with a family history of cancer. Pretest education and counseling should reduce these high-risk perceptions. Better estimates of carrier probability will direct more intensive clinical services and research. PMID- 9742921 TI - Hilar Cholangiocarcinoma: patterns of spread, the importance of hepatic resection for curative operation, and a presurgical clinical staging system. AB - OBJECTIVES: To determine the resectability rate for hilar cholangiocarcinoma, to analyze reasons for unresectability, and to devise a presurgical clinical T staging system. METHODS: Ninety patients with hilar cholangiocarcinomas seen between March 1, 1991, and April 1, 1997, were evaluated. Accurate patterns of disease progression and therapy were evaluable. Disease was staged in 87 patients using extent of ductal tumor involvement, portal vein compromise, and liver atrophy. RESULTS: In 21 patients, disease was deemed unresectable for cure at presentation. In 39 patients, disease was found to be unresectable at laparotomy, 23 secondary to nodal (N2) or distant metastases. Unresectability was the result of metastases in 52% and of locally advanced disease in 28%. Thirty patients (33%) had resection of all gross disease, and 25 of these (83%) had negative histologic margins. Twenty-two patients underwent partial hepatectomy. The 30-day mortality rate was 7%. Projected survival is greater than 60 months in those with a negative histologic margin, with a median follow-up of 26 months. A presurgical T-staging system allows presurgical selection for therapy, predicts partial hepatectomy, and offers an index of prognosis. CONCLUSIONS: In half the patients, unresectability is mainly the result of intraabdominal metastases. Presurgical imaging predicts unresectability based on local extension but is poor for assessing nodal metastases. In one third of patients, disease can be resected for cure with a long median survival. Curative resection depends on negative margins, and hepatic resection is necessary to achieve this. The T-staging system correlates with resectability, the need for hepatectomy, and overall survival. PMID- 9742922 TI - Conservative management of late rejection after heart transplantation: a 10-year analysis. AB - OBJECTIVE: Immunosuppressive regimens for rejection after heart transplantation have been modified to reduce infectious complications without diminishing rejection treatment efficacy. A review of a single institutional series was performed to evaluate the influence of conservative management of grade 2 rejection on long-term outcomes after heart transplantation. METHODS: Before 1990, patients with late (>3 months after transplant) grade 2 rejection were treated with supplemental immunosuppressive drugs. Beginning in 1990, patients with late grade 2 rejection were treated conservatively by maintaining the current immunosuppressive regimen without additional therapy. The groups were compared for survival, incidence of subsequent rejection, and incidence of subsequent infection. RESULTS: One hundred twelve patients had one or more episodes of isolated, late grade 2 rejection; 39 (35%) were treated with supplemental immunosuppression (treated group) and 73 (65%) received no additional therapy (nontreated group). The mean time from transplantation to the first episode of isolated grade 2 rejection was 15.6 months in the treated group and 17.8 months in the nontreated group. Graft survival at 5 and 10 years was 69% and 51 %, respectively, in the treated group and 67% and 41 %, respectively, in the nontreated group (p = 0.77). The rates for overall subsequent rejection were 0.031 episodes/patient-month in the treated group and 0.029 episodes/patient month in the nontreated group (p = 0.64). The rates for early rejection within 6 months of initial grade 2 rejection were 0.044 episodes/patient-month in the treated group and 0.035 episodes/patient-month in the nontreated group (p = 0.56). The rates for overall subsequent infection were 0.018 episodes/patient month in the treated group and 0.012 episodes/patient-month in the nontreated group (p = 0.05). The rates for early infection within 6 months of initial grade 2 rejection were 0.070 episodes/patient-month in the treated group and 0.032 episodes/patient-month in the nontreated group (p = 0.04). Group comparisons demonstrated a significantly lower incidence of infection in the nontreated group. CONCLUSIONS: Conservative management of late grade 2 rejection neither adversely affects survival nor increases the incidence of subsequent short-term or long-term rejection. This approach lowers the early and late incidence of infection after rejection and may reduce other complications from aggressive supplemental immunosuppression. PMID- 9742923 TI - Innominate artery reconstruction: over 3 decades of experience. AB - SUMMARY BACKGROUND DATA: Symptomatic atherosclerotic occlusive disease of the innominate artery is a threatening disease pattern that offers a major challenge in achieving definitive surgical repair. To assess the evolution of treatment strategies and their outcomes for this disease, the authors undertook a review of the cumulative experience for more than 3 decades at one institution. METHODS: Between 1960 and 1997, 94 patients (mean age, 62 years) underwent direct innominate artery revascularization for occlusive atherosclerotic disease to relieve neurologic (n = 85) and/or right upper extremity (n = 26) symptoms or asymptomatic critical stenosis (n = 3). The pattern of atherosclerotic involvement revealed by angiography included critical stenosis (n = 77), complete occlusion (n = 10), and moderate stenosis with plaque ulceration (n = 7). A common brachiocephalic trunk was present in five patients. Transsternal (n = 68) or transcervical (n = 4) innominate endarterectomy was performed in 72 patients and bypass grafting in 22. Forty-one patients underwent concomitant endarterectomy or bypass of innominate branches or adjacent arch vessels, and 3 had coronary bypass grafting. RESULTS: There were three perioperative deaths (3%), all due to cardiac causes. Postoperative morbidity included four strokes (three resolved), two myocardial infarctions, two transient ischemic attacks, and one sternal dehiscence. Follow-up ranged from 8 months to 20 years. Postoperative actuarial survival rate was 96% at 1 year, 85% at 5 years, and 67% at 10 years. Freedom from recurrence requiring reoperation was 100% at 1 year, 99% at 5 years, and 97% at 10 years. CONCLUSIONS: Innominate artery reconstruction is safe and durable when either endarterectomy or prosthetic bypass is used. The anatomic variation and disease distribution permit endarterectomy for most patients. The technique of innominate endarterectomy can be extended safely to outflow and adjacent vessels. PMID- 9742924 TI - Long-term results of pediatric liver transplantation: an analysis of 569 transplants. AB - OBJECTIVE: To analyze a single center's 13-year experience with 569 pediatric orthotopic liver transplants for end-stage liver disease. SUMMARY BACKGROUND DATA: Despite advances in medical therapy, liver replacement continues to be the only definitive mode of therapy for children with end-stage liver disease. Innovative surgical techniques and improved immunosuppression have broadened the application of liver replacement for affected children. However, liver transplantation in the child remains challenging because of the scarcity of donor organs, complex surgical technical demands, and the necessity to prevent long term complications. METHODS: The medical records of 440 consecutive patients younger than 18 years of age undergoing orthotopic liver transplantation for end stage liver disease from March 20, 1984, to November 15, 1997, were reviewed. Results were analyzed using Cox multivariate regression analysis to determine the statistical strength of independent associations between pretransplant covariates and patient and graft survival. Actuarial patient and graft survival rates were determined at 1, 3, 5, and 10 years. The type and incidence of posttransplant complications were determined, as was the quality of long-term allograft function. The median follow-up period was 4.1 years. RESULTS: Biliary atresia was the most common cause (50.4%) of endstage liver disease in this patient population. The median recipient age was 2.4 years; 239 patients (54%) were younger than 3 years of age and 1 11 patients (25%) were younger than 1 year of age. There were 471 whole organs, 29 were ex vivo reduced size, 33 were living related donor, and 36 were in situ split-liver allografts. Three hundred forty three (78%) patients underwent a single allograft, whereas 97 patients required retransplantation; hepatic artery thrombosis was the most common indication for retransplantation (55 patients). The 1-, 3-, 5-, and 10-year actuarial patient survival rates were 82%, 80%, 78%, and 76%, respectively; allograft survival rates were 68%, 63%, 60%, and 54%. Long-term liver function remains excellent: current median follow-up values for total bilirubin and aspartate aminotransferase were 0.5 mg/dl and 54 IU/L, respectively. Cox multivariate regression analysis demonstrated that pretransplant patient age, the era of transplantation, and the number of allografts performed significantly and independently predicted patient survival rates, whereas allograft type and pretransplant diagnosis did not. CONCLUSIONS: Liver transplantation in the pediatric patient is a durable procedure that provides excellent long-term survival. Although there have been overall improvements in patient outcome with increased experience, the effect is most pronounced for patients younger than 1 year of age. Retransplantation, although effective in a meaningful number of patients, continues to carry a progressive decrement in survival with the number of allografts performed. Use of living-related and in situ split-liver allografts has dramatically reduced waiting times for small children and has improved patient survival. PMID- 9742925 TI - Kidney transplantation in children younger than 1 year using cyclosporine immunosuppression. AB - OBJECTIVE: The optimal age for transplantation in children with end-stage renal disease remains controversial. Supported by national data, many centers recommend dialysis until the child reaches a certain minimum age. The authors' policy, however, has been to encourage living donor (LD) transplants for young children, with no minimum age restriction. METHODS: Between January 1, 1984, and December 31, 1996, the authors performed 248 kidney transplants in children younger than age 13 years, using cyclosporine as the primary immunosuppressive agent. Recipients were analyzed in three age groups: group 1, younger than age 1 year (n = 26); group 2, age 1 through 4 (n = 92); and group 3, age 5 through 13 (n = 130). Almost all recipients in group 1 underwent a primary LD transplant. Therefore, to compare results more meaningfully among the three age groups, only primary LD transplants were analyzed (group 1, n = 25; group 2, n = 59; group 3, n = 58). RESULTS: In primary LD transplants, no significant difference was noted among the age groups in 1-and 5-year patient or graft survival rates. To date, all 25 recipients from group 1 are alive and well; 19 still have a functional original graft. Causes of graft loss in the remaining six recipients were chronic rejection (n = 3), vascular thrombosis (n = 2), and recurrent disease (n = 1). The incidence of acute rejection in group 1 recipients was lower than in the two older groups. However, the incidence of delayed graft function was slightly higher in the youngest group than in the two older groups. For recipients in group 1, growth (as measured by weight) improved significantly posttransplant: the mean standard deviation score rose from -2.8 pretransplant to -0.2 by age 5 and to +1.8 by age 10. The improvement in height was not as dramatic: the mean standard deviation score rose from -3.2 pretransplant to -1.6 by age 5 and to 1.4 by age 10. CONCLUSIONS: Kidney transplantation in young children, including those younger than 1 year old, can achieve results comparable to those in older children. As long as an adult LD is available, the timing of the transplant should be based on renal function rather than age. PMID- 9742927 TI - Distribution and functional diversification of the ras superfamily in Saccharomyces cerevisiae. AB - The recent availability of the full Saccharomyces cerevisiae genome sequence offers a first opportunity to analyze the composition, function and evolution of GTPases in the ras-p21 superfamily. This superfamily in yeast is composed of 29 proteins divided into five families: ras with four sequences implicated in cell signalling; rho, six genes related to the cell shape machinery; ypt-rab, ten proteins with different roles in intracellular trafficking; arf-sar, seven proteins related to vesicular trafficking in secretory pathways; and ran, two proteins acting as components of the nuclear transport system. The superfamily covers a wide range of cellular functions from signalling to intracellular trafficking, while conserving the structural framework and a common mechanism of GTP hydrolysis. PMID- 9742926 TI - Importance of hospital volume in the overall management of pancreatic cancer. AB - OBJECTIVE: To determine whether hospital volume is associated with clinical and economic outcomes for patients with pancreatic cancer who underwent pancreatic resection, palliative bypass, or endoscopic or percutaneous stent procedures in Maryland between 1990 and 1995. SUMMARY BACKGROUND DATA: Previous studies have demonstrated that outcomes for patients undergoing a Whipple procedure improve with higher surgical volume, but only 20% to 35% of patients with pancreatic cancer qualify for curative resection. Most patients undergo palliative procedures instead with a surgical bypass or biliary stent. METHODS: Analysis of hospital discharge data from all nonfederal acute care hospitals in Maryland identified all patients with pancreatic cancer who underwent a pancreatic resection, palliative bypass, or stent procedure between 1990 and 1995. Hospitals (n = 48) were categorized as high-, medium-, and low-volume providers according to their average annual volume of these procedures. Multivariate regression was used to examine the association between hospital volume and in-hospital mortality rate, length of stay, and hospital charges, after adjusting for differences in case mix and surgeon volume. RESULTS: Increased hospital volume is associated with markedly decreased in-hospital mortality rates and a decreased or similar length of stay for all three types of procedures and with decreased or similar hospital charges for resections and stents. After adjustment for case mix differences, the relative risk (RR) of in-hospital death after pancreatic resection was 19.3 and 8 at the low- and medium-volume hospitals, respectively, versus the high-volume hospital; after bypasses, the RR of death was 2.7 and 1.9, respectively; and after stents, the RR was 4.3 and 4.8, respectively. CONCLUSIONS: Patients with pancreatic cancer who are to be treated with curative or palliative procedures appear to benefit from referral to a high-volume provider. PMID- 9742928 TI - Intracellular localization and release of eotaxin from normal eosinophils. AB - Eotaxin is a potent and selective CC chemokine for eosinophils and basophils. We established several monoclonal antibodies (Mabs) allowing the neutralization and measurement of human eotaxin. Using the Mabs as probes, we demonstrated that normal eosinophils contained intracellular granule-associated eotaxin. Quantification of cell-associated eotaxin in different leukocyte subsets revealed that it was principally expressed in eosinophils. Finally, we showed that normal eosinophils released eotaxin upon stimulation with either of two secretagogues, C5a or ionomycin. These findings raise the possibility that eosinophil-derived eotaxin contributes to the local accumulation of eosinophils at the site of inflammation. PMID- 9742929 TI - Biosynthesis of D-aspartate in mammalian cells. AB - In this communication, we demonstrate that D-aspartate (D-Asp) is synthesized in pheochromocytoma cells (PC12). To our knowledge this is the first report of biosynthesis of D-Asp in mammalian cells. Synthesis of D-Asp was demonstrated by its time-dependent accumulation in the cell culture, and by the fact that this accumulation was proportional to the number of inoculated cells. D-Asp in PC12 cells was identified by (i) co-elution with authentic D-Asp on two different HPLC columns, an octadesyl silica column and a Pirkle-type chiral column, (ii) reversed elution order of D-Asp and L-Asp on another Pirkle-type chiral column with an opposite configuration, and (iii) sensitivity to D-Asp oxidase. In the cells the amount of D-Asp was approx. 12-14% of total Asp and no other investigated D-amino acid was detected. The amount of D-Asp did not increase during the culture of mouse 3T3 fibroblasts and human neuroblastoma NB-1 cells. Immunocytochemical staining with anti-D-Asp antiserum demonstrated that D-Asp synthesized is present in the cytoplasm of the cells. PMID- 9742930 TI - Inhibition of voltage-gated cationic channels in rat embryonic hypothalamic neurones and C1300 neuroblastoma cells by triphenylethylene antioestrogens. AB - The effect of the non-steroidal antioestrogens tamoxifen and toremifene on voltage-gated cationic currents was examined in primary cultures of rat hypothalamic neurones and the C1300 mouse neuroblastoma cell line using the whole cell patch clamp technique. When applied to the external bathing solution both tamoxifen and toremifene were able to inhibit TTX-sensitive sodium currents with IC50 values of 1-2 microM and delayed rectifier type potassium currents (IC50, 2 3 microM). However, only toremifene showed a significant inhibition of the I(A) current (IC50 3 microM). Inhibition of voltage-gated cationic currents was significantly impaired when tamoxifen was applied in a serum-containing solution. The steroidal antioestrogen ICI 182,780 did not inhibit any of the currents at 10 microM. PMID- 9742931 TI - Pervanadate-triggered MAP kinase activation and cell proliferation are not sensitive to PD 98059. Evidence for stimulus-dependent differential PD 98059 inhibition mechanism. AB - A tight and stable complex with corresponding protein kinases and phosphatases establishes coupling between activators and inactivators. One such example is emerging from the studies of the Ras-dependent MAP kinase cascade signaling pathway. Pervanadate, a potent inhibitor of protein tyrosine phosphatase, stimulates MAP kinase and elicits cell proliferation in cultured mouse fibroblasts which is insensitive to PD 98059, the major inhibitor of upstream MEK, whereas serum- or TPA-triggered proliferation is sensitive to PD 98059. It is suggested that imbalanced coordination between protein kinase and protein phosphatase determines the cellular responses such as cell proliferation. The PD 98059-insensitive cell proliferation upon protein tyrosine phosphatase inhibition is attributed to a MEK bypass pathway. PMID- 9742932 TI - Stimulation by anaphylatoxin C5a of glycogen phosphorylase in rat hepatocytes via prostanoid release from hepatic stellate cells but not sinusoidal endothelial cells. AB - In the perfused rat liver, the anaphylatoxin C5a has been shown to enhance glucose output. Since hepatocytes lack C5a receptor mRNA, the metabolic effect of C5a must be elicited indirectly via C5a receptor expressing non-parenchymal liver cells. Kupffer cells were found to be able to mediate the C5a action via release of prostanoids. However, elimination of the Kupffer cells by pretreatment of the animals with gadolinium chloride reduced the metabolic effect of C5a to only about 40%. Therefore, it was investigated whether not only Kupffer cells but in addition also hepatic stellate cells or sinusoidal endothelial cells released prostanoids in response to C5a. In isolated hepatic stellate cells but not in sinusoidal endothelial cells, recombinant rat C5a induced a time- and dose dependent release of thromboxane B2 and prostaglandins D2, E2 and F2alpha. The rate of prostanoid release was maximal within the first two minutes and then declined again. C5a-induced prostanoid release from hepatic stellate cells was smaller than that from Kupffer cells and it differed in the prostanoid ratios (PGE2/PGD2/PGF2alpha/TXB2 = 1:1:0.1:0.6 and 1:4:1:3, respectively). RrC5a activated hepatocellular glycogen phosphorylase via prostanoid release in cocultures of hepatocytes with hepatic stellate cells but not with sinusoidal endothelial cells. Thus, the part of the rrC5a-induced glucose output in the perfused rat liver, which was not abrogated by elimination of the Kupffer cells with gadolinium chloride, most likely was mediated by prostanoids released from hepatic stellate cells. PMID- 9742933 TI - An AP-1 site in the promoter of the human IL-5R alpha gene is necessary for promoter activity in eosinophilic HL60 cells. AB - Interleukin-5 (IL-5) plays a crucial role in the proliferation, differentiation and activation of eosinophils. The IL-5 receptor is composed of an IL-5-specific alpha subunit, which is expressed by eosinophils and basophils, and a beta c subunit shared with the receptors for IL-3 and GM-CSF. We identified an AP-1 element which is important for IL-5R alpha promoter activity in eosinophilic HL60 cells. The AP-1 site and the previously identified EOS1 site cooperate, since single mutation of either of the sites decreased promoter activity. We show that the AP-1 site of the IL-5R alpha promoter binds multiple proteins, including cJun, CREB, and CREM. PMID- 9742934 TI - Engineering of hypoallergenic mutants of the Brassica pollen allergen, Bra r 1, for immunotherapy. AB - The Brassica pollen allergen Bra r 1 belongs to a new family of Ca2+-binding proteins, characterized by the presence of two potential EF-hand calcium-binding domains. Disruption of these EF-hand motifs by amino acid substitutions demonstrated that both domains of Bra r 1 constitute functional Ca2+-binding sites. Calcium-binding deficient mutants displayed significantly reduced IgE binding activity. Injection of these mutated Bra r 1 variants into a murine model system showed that mouse IgG raised against the mutants recognized native Bra r 1 in Brassica pollen extracts suggesting the potential use of the engineered allergens for effective immunotherapy. PMID- 9742935 TI - Tetrahydrobiopterin-dependent stabilization of neuronal nitric oxide synthase dimer reduces susceptibility to phosphorylation by protein kinase C in vitro. AB - Binding of (6R)-5,6,7,8-tetrahydro-L-biopterin (H4B) stabilizes the homodimeric structure of neuronal nitric oxide synthase (nNOS). In the present study, low temperature sodium dodecylsulfate-polyacrylamide gel electrophoresis revealed differential susceptibility of stabilized and non-stabilized dimers to in vitro phosphorylation by protein kinase C. Protein kinase C preferentially phosphorylated the non-stabilized dimer. Although a low extent of phosphorylation was detected in the stabilized dimer, most of it was estimated to be due to phosphorylation of the dimer before its stabilization. Phosphorylation did not affect the stabilizing effect of H4B. These results indicate that H4B-dependent dimer stabilization prevents nNOS from protein kinase C-dependent phosphorylation in vitro. PMID- 9742936 TI - The M2 channel of influenza A virus: a molecular dynamics study. AB - Molecular dynamics simulations have been performed on a tetramer of the 25 residue (SSDPLVVAASIIGILHLILWILDRL) synthetic peptide [1] which contains the transmembrane domain of the influenza A virus M2 coat protein. The peptide bundle was initially assembled as a parallel alpha-helix bundle in the octane portion of a phase separated water/octane system, which provided a membrane-mimetic environment. A 4-ns dynamics trajectory identified a left-handed coiled coil state of the neutral bundle, with a water filled funnel-like structural motif at the N-terminus involving the long hydrophobic sequence. The neck of the funnel begins at V27 and terminates at H37, which blocks the channel. The C-terminus is held together by inter-helix hydrogen bonds and contains water below H37. Solvation of the S23 and D24 residues, located at the rim of the funnel, appears to be important for stability of the structure. The calculated average tilt of the helices in the neutral bundle is 27 +/- 5 degrees, which agrees well with recent NMR data. PMID- 9742937 TI - Isolation of human intestinal defensins from ileal neobladder urine. AB - We describe the isolation of naturally occurring human intestinal defensins HD-5 and HD-6 from ileal neobladder urine and ileal mucosa. Using an antibody-based detection assay, we found multiple N-terminally processed forms of HD-5. The predominant HD-5 forms in tissue were longer than those in neobladder urine (amino acid (aa) 23-94 and 29-94 versus aa 36-94, 56-94 and 63-94) suggesting that Paneth cells store prodefensin that is processed to mature defensin during or after degranulation. Search for mature HD-6 yielded aa 69-100 as the predominant form in both sources. The ileal neobladder is a promising model to study human Paneth cell secretion. PMID- 9742938 TI - The GalR2 galanin receptor mediates galanin-induced jejunal contraction, but not feeding behavior, in the rat: differentiation of central and peripheral effects of receptor subtype activation. AB - The neuropeptide galanin mediates a diverse array of physiological functions through activation of specific receptors. Roles of the three recently cloned galanin receptors (GalRs) in rat intestinal contraction and food intake were examined using GalR-selective ligands and the results were compared with the pharmacological profiles of defined GalRs. The action profile of these ligands in jejunal contraction resembled only that of GalR2 and only a high level of GalR2 mRNA was detected in the tissue, supporting GalR2 as the receptor mediating jejunal contraction. The action profile for food intake in rats excluded GalR2, GalR3 and the putative pituitary galanin receptor as the 'feeding receptor', suggesting that either GalR1 or an unidentified GalR is responsible for mediating this function. PMID- 9742939 TI - Soluble IL-1 receptor type I binds to human dermal fibroblasts and induces calcium flux. AB - Soluble cytokine receptors appear to modify ligand concentrations by stabilizing ligands or by specifically inhibiting interactions of ligands with their membrane bound receptors. Here we describe a new function of the soluble interleukin-1 receptor type I (IL-1sR I). This receptor induced a transient rise of intracellular free calcium concentration in human dermal fibroblasts in a dose dependent fashion. Mobilization of calcium by IL-1sR I was abolished in the presence of an equimolar concentration of IL-1 receptor antagonist (IL-1ra). Neutralizing antibodies against IL-1beta also abolished calcium mobilization stimulated with IL-1sR I indicating that IL-1beta is involved. IL-1sR I bound with high affinity (Kd 1-2 nM) to the fibroblasts. In addition, IL-1sR I enhanced expression of IL-6 and IL-8 mRNA. The observation that IL-1sR I can act as a ligand and agonist for membrane IL-1 extends the concept of the ligand-receptor functions of both IL-1 and IL-1sR I and adds a new dimension to the cytokine network. PMID- 9742940 TI - A new human nm23 homologue (nm23-H5) specifically expressed in testis germinal cells. AB - We have identified a cDNA encoding a 212 amino acid protein (Nm23-H5) with 27-31% identity to the human members of the nm23/nucleoside diphosphate (NDP) kinase gene family. The nm23-H5 gene is located on chromosome 5q23-31 and is transcribed as one main transcript of 1.1 kb which is highly and specifically expressed in testis, in the spermatogonia and early spermatocytes. Nm23-H5 possesses most of the residues conserved among NDP kinases plus an additional COOH-terminus end of 55 amino acids unique to this protein. However, under usual assay conditions, Nm23-H5 expressed in Escherichia coli did not exhibit NDP kinase activity. These results suggest that Nm23-H5 is specifically involved in early stages of spermatogenesis. PMID- 9742942 TI - Expression profile and structural divergence of novel human annexin 31. AB - Systematic analysis of expressed sequence tags in dbEST yielded an expression profile of the ten known human annexins and led to the discovery of a novel subfamily expressed mainly in differentiating tissues. Full-length cDNAs encoded a 338-amino acid protein with less than 40% identity to other annexins, an atypical amino acid composition, and an insertion and deletion in internal repeat 3. The most striking feature was a complete ablation of all four type II calcium binding sites in the conserved tetrad core. Annexin 31 thus constitutes a unique, natural probe for investigating the role of membrane binding in annexin function. PMID- 9742941 TI - On the expression of cytosolic calcium-independent phospholipase A2 (88kDa) in immature and mature myeloid cells and its role in leukotriene synthesis in human granulocytes. AB - The human calcium-independent phospholipase A2 (iPLA2; 88 kDa) has recently been cloned (Larsson, P.K.A., Claesson, H.-E. and Kennedy, B.P. (1998) J. Biol. Chem. 272, 207-214). Here we demonstrate the expression of the human iPLA2 mRNA and its splice variants in blood progenitor cells, immature leukemic cells and mature granulocytes. Chromatographical resolvable iPLA2 activity was found in the cytosolic fraction of granulocytes and the activity was inhibited by the iPLA2 inhibitor bromoenol lactone. This drug also inhibited leukotriene synthesis in human granulocytes, induced by low concentration of calcium ionophore A23187 (0.10-0.15 microM) or opsonized zymosan. These results suggest that iPLA2 is involved in the regulation of the pool of arachidonic acid destined for leukotriene synthesis in human granulocytes. PMID- 9742943 TI - Activation of mitogen-activated protein kinases (p38-MAPKs, SAPKs/JNKs and ERKs) by adenosine in the perfused rat heart. AB - Adenosine and mitogen-activated protein kinases (MAPKs) have been separately implicated in cardiac ischaemic preconditioning. We investigated the activation of MAPK subfamilies by adenosine in perfused rat hearts. p38-MAPK was rapidly phosphorylated and activated (10-fold activation, maximal at 5 min) by 10 mM adenosine, as was the p38-MAPK substrate, MAPKAPK2 (4.5-fold). SAPKs/JNKs were activated (5-fold) and ERKs were phosphorylated (both maximal at 5 min). The concentration dependences of activation of p38-MAPK and ERKs were biphasic with a 'high affinity' component (maximal at 10-100 microM adenosine) and a 'low affinity' component that had not saturated at 10 mM. SAPKs/JNKs were activated only by 10 mM adenosine. These results are consistent with MAPK involvement in adenosine-mediated ischaemic preconditioning. PMID- 9742944 TI - Observing the deoxy myoglobin and hemoglobin signals from rat myocardium in situ. AB - 1H NMR proximal histidyl NdeltaH signals of deoxy hemoglobin (Hb) and myoglobin (Mb) are distinguishable in the rat myocardium in situ. In the normoxic resting state, the blood and tissue pO2 is sufficient to saturate both Mb and Hb. No deoxy Mb or Hb signals are detected. Under 12% inspired O2, the erythrocyte Hb is partially desaturated and yields the alpha and beta proximal histidyl NdeltaH signals of deoxy Hb. The detection of the Hb signals clarifies the debate about the NMR visibility of erythrocyte Hb in vivo and augments the strategy to observe tissue pO2. PMID- 9742945 TI - The permeability transition pore induced under anaerobic conditions in mitochondria energized with ATP. AB - The role of oxygen in the induction of mitochondrial permeability transitions was studied. Oxygen consumption, swelling, membrane potential and calcium transport were recorded simultaneously in isolated rat liver mitochondria. Oxygen depletion was accomplished by saturating the medium with N2 and allowing either mitochondrial respiration or glucose/glucose oxidase to consume the residual oxygen. Upon anaerobiosis, mitochondria were supplemented with 500 microM ATP to support succinate-driven membrane potential. Under these conditions, 100 microM Ca2+ induced cyclosporin A-sensitive permeability transitions. To eliminate the possible inhibition of permeability transition by high concentrations of adenine nucleotides, anaerobic mitochondria were also energized by the combination of 20 microM ADP and phosphoenolpyruvate/pyruvate kinase. These mitochondria also underwent Ca2+-induced permeability transition. Under both of these conditions, namely the addition of ATP as a single or through actions of pyruvate kinase, the respiratory components were totally reduced. Thus, oxygen is not a necessary factor for mitochondria to undergo permeability transitions. PMID- 9742947 TI - Electron entry in a CuA mutant of cytochrome c oxidase from Paracoccus denitrificans. Conclusive evidence on the initial electron entry metal center. AB - A cytochrome c oxidase subunit II C216S mutant from Paracoccus denitrificans in which the CuA site was changed by site-directed mutagenesis to a mononuclear copper site [Zickermann, V., Wittershagen, A., Kolbesen, B.O. and Ludwig, B. Biochemistry 36 (1997) 3232-3236] was investigated by stopped-flow spectroscopy. Contrary to the behavior of the wild type enzyme, in this mutant cytochrome a cannot be reduced by excess cytochrome c in the millisecond time scale in which cytochrome c oxidation is observed. The results conclusively identify and establish CuA as the initial electron entry site in cytochrome c oxidase. Partial rapid reduction (ca. 20%) of the modified CuA site suggests that the mononuclear copper ion has a redox potential ca. 100 mV lower than the wild type, and that internal electron transfer to cytochrome a is > or = 10(3)-fold slower than with the wild type enzyme. PMID- 9742946 TI - Human serum, cysteine and histidine inhibit the oxidation of low density lipoprotein less at acidic pH. AB - Low concentrations of serum or interstitial fluid have been shown to inhibit the oxidation of low density lipoprotein (LDL) catalysed by copper or iron, and may therefore protect against the development of atherosclerosis. As atherosclerotic lesions may have an acidic extracellular pH, we have investigated the effect of pH on the inhibition of LDL oxidation by serum and certain components of serum. Human serum (0.5%, v/v), lipoprotein-deficient human serum at an equivalent concentration and the amino acids L-cysteine (25 microM) and L-histidine (25 microM), but not L-alanine (25 microM), inhibited effectively the oxidation of LDL by copper at pH 7.4, as measured by the formation of conjugated dienes. The antioxidant protection was reduced considerably at pH 6.5, and was decreased further at pH 6.0. These observations may help to explain why LDL becomes oxidised locally in atherosclerotic lesions in the presence of the strong antioxidant protection offered by extracellular fluid. PMID- 9742948 TI - Structure of the Na+-driven flagellum from the homoacetogenic bacterium Acetobacterium woodii. AB - The Na+-dependent flagellum of Acetobacterium woodii was characterised. Flagellin and whole flagella were purified and analysed by SDS-PAGE and electron microscopy. The structure and dimensions of the filament and the hook-basal body, as revealed by electron microscopy, resemble those of H+-dependent flagella from gram-positive bacteria. Intramembrane particle rings were present at the cell pole in freeze-fractured A. woodii cells, which might correspond to the mot complex. PMID- 9742949 TI - Insulin-like growth factor-binding protein-2 inhibits proliferation of human embryonic kidney fibroblasts and of IGF-responsive colon carcinoma cell lines. AB - So far, the physiological role of insulin-like growth factor binding protein-2 (IGFBP-2) has not been demonstrated directly. Therefore, we transfected 293 cells with an expression vector containing the CMV promoter and the complete cDNA of mouse IGFBP-2. Secretion of bioactive IGFBP-2 into conditioned medium was demonstrated by Western ligand and Western immunoblotting and quantified by specific RIA. For the analysis of cell proliferation three clones exhibiting either high or low/no IGFBP-2 expression were selected and compared to non transfected parental 293 cells. IGFBP-2 secreting clones displayed reduced conversion of thiazolyl blue when compared to negative clones or non-transfected parental 293 cells (P < 0.01). The lower growth activity measured in the IGFBP-2 secreting clones was compensated in great part by the administration of exogenous IGF-I or -II. Conditioned media of IGFBP-2 secreting clones inhibited growth of IGF-responsive colon tumor cell lines (LS513, HT-29) while those of negative clones did not. In addition, conditioned medium from a clone expressing high levels of IGFBP-2 inhibited anchorage-independent growth of LS513 and HT-29 cells. In contrast, growth of an IGF-unresponsive tumor cell line (Co-115) was not affected by the conditioned media. We hypothesize that IGFBP-2 might sequester the IGFs and thus prevent them from transferring their mitogenic signals. PMID- 9742950 TI - Evidence for the archaebacterial-type conformation about the bond between the beta-ionone ring and the polyene chain of the chromophore retinal in chlamyrhodopsin. AB - Previous studies using retinal analogs (Wada, A., Sakai, M., Kinumi, T., Tsujimoto, K., Yamauchi, M. and Ito, M. (1994) J. Org. Chem. 59, 6992-6997; Wada, A., Sakai, M., Imamoto, Y., Shichida., Y., Yoshizawa, T. and Ito, M. (1993) Chem. Pharm. Bull. 41, 793-795) revealed that both retinochrome and visual pigments share a common chromophoric conformation in which the ring region of retinal is twisted ca. 50 degrees with respect to the plane of the polyene chain, suggesting a highly conserved 6-s-cis conformation throughout rhodopsin-like pigments in animals. By contrast, 6-s-trans conformation was commonly observed or suggested in archaebacterial rhodopsins examined thus far. Here we have reconstituted in vivo both the photoreceptor for photobehavioral responses of the unicellular alga Chlamydomonas reinhardtii and the second archaeal sensory photoreceptor phoborhodopsin from a series of retinal analogs. Results exclusively point to the conclusion that in both photoreceptors retinal has the coplanar 6-s-trans conformation, though recent molecular cloning studies revealed no homology between Chlamydomonas photoreceptor (chlamyrhodopsin) and archaeal rhodopsins. PMID- 9742951 TI - Tyrosine phosphorylation of the TATA element modulatory factor by the FER nuclear tyrosine kinases. AB - The FER locus in the mouse encodes two tyrosine kinases, p94fer and p51ferT. While p94fer accumulates in the cytoplasm and nucleus of most mammalian cells the expression of p51ferT is restricted to the nucleus of meiotic primary spermatocytes. The cellular function of the FER kinases is not understood, nor has a substrate for these enzymes been characterized. To identify putative substrates of p94fer and p51ferT, the two enzymes were used as 'baits' in the yeast two-hybrid screening system. cDNAs encoding the mouse TATA element modulatory factor (TMF) were repeatedly isolated in this assay. TMF was previously shown to bind the TATA element in RNA polymerase II promoters and impaired their functioning in a cell free transcription system. Both p94fer and p51ferT phosphorylated the TMF protein in in vitro and in vivo kinase assays. Sequential deletions showed that the carboxy-terminal region of TMF was essential for phosphorylation. In situ hybridization analysis revealed the preferential accumulation of TMF transcripts in meiotic spermatogenic and oogenic cells. p94fer and p51ferT may thus modulate the suppressive activity of TMF during cellular growth and in defined differentiation processes. PMID- 9742952 TI - Caloric restriction prevents age-related decline in skeletal muscle dihydropyridine receptor and ryanodine receptor expression. AB - The dihydropyridine receptor (DHPR), a voltage-gated L-type Ca2+ channel, and the Ca2+ release channel/ryanodine receptor isoform-1 (RyR1) are key molecules involved in skeletal muscle excitation-contraction coupling. We have reported age related decreases in the level of DHPR expression in fast- and slow-twitch muscles from Fisher 344 cross Brown Norway (F344BNX) rats (Renganathan, Messi and Delbono, J. Membr. Biol. 157 (1997) 247-253). Based on these studies we postulate that excitation-contraction uncoupling is a basic mechanism for the decline in muscle force with aging (Delbono, Renganathan and Messi, Muscle Nerve Suppl. 5 (1997) S88-92). In the present study, we extended our studies to older ages and we intended to prevent or retard excitation-contraction uncoupling by restricting the caloric intake of the F344BNX rats from 16 weeks of age. Three age groups, 8 , 18-, and 33-month old caloric restricted rats, were compared with ad libitum fed animals. The number of DHPR and RyR1 and DHPR/RyR1 ratio (an index of the level of receptors uncoupling) in skeletal muscles of 8-month and 18-month rats was not significantly different in either ad libitum fed or caloric restricted rats. However, the age-related decrease in the number of DHPR, RyR1 and DHPR/RyR1 ratio observed in 33-month old ad libitum fed rats was absent in 33-month old caloric restricted rats. These results suggest that caloric restriction prevents age-related decreases in the number of DHPR, RyR1 and DHPR/RyR1 ratio observed in fast- and slow-twitch rat skeletal muscles. PMID- 9742953 TI - pH-sensitive liposomes for receptor-mediated delivery to chicken hepatoma (LMH) cells. AB - pH-sensitive liposomes composed of dioleoylphosphatidylethanolamine and cholesterol hemisuccinate (3:2 mol/mol) bearing the N-acetylglucosamine derivative of bovine serum albumin (N-Ac-BSA) were applied for receptor-mediated delivery in chicken hepatoma (LMH) cells expressing the N-Ac-BSA-binding asialoglycoprotein receptor. Fluorescently labeled dextran was entrapped in liposomes by a modified freeze-thawing method (encapsulation efficiency of 23%). A novel method of coupling proteins onto the surface of preformed liposomes yielded a coupling efficiency of 60-70%. The association of pH-sensitive and lecithin liposomes with LMH cells was monitored by fluorescence-activated cell sorting and confocal microscopy. Prerequisites for receptor-mediated delivery to LMH cells were both the pH sensitivity of liposomes and the presence of N-Ac-BSA on the liposomal surface. PMID- 9742954 TI - Decelerated degradation of short peptides by the 20S proteasome. AB - Based on a twelve residue master peptide comprising all five specific cleavage sites defined for the proteasome, a set of variant peptides was generated in order to probe specificity and to elucidate the mechanism which determines product size. It is shown that the rate of degradation by the 20S proteasome from Thermoplasma acidophilum depends critically on the length of the peptide substrate. Peptides of 14 residues and longer are degraded much faster than shorter peptides although the sites of cleavage remain unchanged. The decelerated degradation of peptides shorter than 14 residues explains the accumulation of products with an average length of seven to nine residues. PMID- 9742955 TI - Oxidative decomposition of malonic acid as basis for the action of manganese peroxidase in the absence of hydrogen peroxide. AB - Manganese peroxidase (MnP) from the ligninolytic basidiomycetes Phlebia radiata and Nematoloma frowardii was found to decompose malonate oxidatively in the absence of H2O2 in a reaction system consisting of the enzyme, sodium malonate and MnCl2. The enzymatic oxidation resulted in a substantial decrease in malonate concentration and the formation of CO2, oxalate, glyoxylate and formate. Simultaneously with the decomposition of malonate, Mn(II) was oxidized to Mn(III) leading to high transient concentrations of the latter. MnP action in the absence of H2O2 started slowly after a lag period of 3 h. The lag period was considerably shortened after a single addition of Mn(III). Superoxide dismutase and catalase inhibited the enzymatic reaction partly, ascorbate completely. ESR studies demonstrated the formation of a carbon-centered radical during the course of the reaction. We propose that the latter generates peroxides that can be used by MnP to oxidize Mn(II) to Mn(III). PMID- 9742956 TI - The extended packaging sequence of MoMLV contains a constitutive mRNA nuclear export function. AB - The present report shows that incorporation of defined sequences from the Moloney murine leukaemia virus (MoMLV) into Rex dependent expression vectors based on the human T-cell leukaemia virus (HTLV-1) allows Rex independent gene expression. Deletion mutagenesis of the MoMLV derived sequences allowed this function to be localised to a 312 nt length sequence overlapping the MoMLV gag p15/p12 open reading frame. This 'extended packaging sequence' has been reported to markedly increase the titre of in vitro packaged retroviral vectors. Using fluorescent in situ hybridisation combined with confocal microscopy we show that the 312 nt element can replace Rex mediated nuclear export and expression of transcripts containing HTLV-1 cis acting repressive elements. Our observations are consistent with the extended packaging sequence of MoMLV exerting a constitutive mRNA nuclear export function. PMID- 9742957 TI - In vitro heat effect on heterooligomeric subunit assembly of thermostable indolepyruvate ferredoxin oxidoreductase. AB - Indolepyruvate ferredoxin oxidoreductase (IOR) from hyperthermophilic archaeon Pyrococcus kodakaraensis KOD1 catalyzes the oxidative decarboxylation of arylpyruvates by forming a heterooligomeric complex (alpha2beta2). The genes iorA and iorB which encode respective alpha and beta subunits, were coexpressed heterologously in Escherichia coli cells under anaerobic conditions. IOR activity was detected from the cell extract containing both subunits and its activity was enhanced by in vitro heat treatment prior to the assay. The iorA and iorB were expressed individually and each subunit was examined for enzymatic activity with and without heat treatment. IOR activity was detected neither from the extract of alpha subunit nor beta subunit. The alpha and beta subunits were mixed and then IOR activity was examined. Weak IOR activity was detected without heat treatment, however, upon heat treatment its activity was enhanced. The mixture of individually heat treated alpha and beta subunits did not possess any IOR activity even though the mixed sample was heat treated again. IOR alpha and beta subunits were individually purified to homogeneity, mixed with or without heat treatment and subunit assembly was examined by determining molecular mass. Upon heat treatment, inactive alpha and beta were converted to an active high molecular weight complex (195 kDa) which corresponds to the alpha2beta2 structure. However, the active complex was not formed without heat treatment, suggesting that high temperature environments are important for the heterooligomerization of IOR subunits. PMID- 9742958 TI - The peptide HDEF as a new retention signal is necessary and sufficient to direct proteins to the endoplasmic reticulum. AB - The key feature of tomato RNase LX localised solely outside the vacuole is the C terminal peptide HDEF which is very similar to known endoplasmic reticulum (ER) retention signals. For functional testing of the ER-targeting ability of HDEF, different constructs including the complete RNase LX, two truncated forms without HDEF and the truncated chitinase FB7-1deltaVTP C-terminally flanked by HDEF, were expressed in Saccharomyces cerevisiae. The majority of RNase and chitinase, both containing HDEF, accumulates within the ER. However, the truncated constructs without the peptide are released into the medium. We provide compelling evidence that peptide HDEF at the C-terminus of secretory plant proteins is a new ER retention signal in yeast and most likely in plants. PMID- 9742959 TI - Isolation and characterization of a rice cDNA encoding the gamma-subunit of translation elongation factor 1B (eEF1Bgamma). AB - We isolated a rice cDNA clone (refg) encoding the gamma-subunit of translation elongation factor 1B (eEF-1B gamma; the old designation was EF-1 gamma). The refg encodes an open reading frame of 419 amino acids which shows a similarity to the equivalent sequences from animals and yeast. Complex formation analysis, which showed the recombinant protein of refg (His-eEF1B gamma) and formed a complex with GST-eEF-1Bbeta, indicated that the refg encodes rice eEF1B gamma of the eEF1B alphabeta gamma complex. Expression analysis showed that refg mRNA is very abundant in suspension-cultured cells during the exponential phase of growth. A DNA blot analysis indicated that refg is located at a single locus in the rice genome. PMID- 9742960 TI - Characterization of early induced genes in Arabidopsis thaliana responding to bacterial inoculation: identification of centrin and of a novel protein with two regions related to kinase domains. AB - The early response to bacterial inoculation has been investigated and two Arabidopsis genes, ap3.3a and ap4.3a have been characterized. The AP3.3A protein showed high identity to centrin, a ubiquitous cytoskeletal protein first identified in unicellular green alga. Amino-acid sequence analyses of the AP4.3A protein indicates that the second gene characterized encodes an unusual protein with two putative kinase domains. Expression of ap3.3a and ap4.3a was rapidly induced after pathogen inoculation. A role of ap3.3a in plant defense could be postulated based on its preferential induction during the incompatible interactions analyzed. In contrast, activation of ap4.3a was not specific and could be related to a more general stress response. PMID- 9742961 TI - Peroxisome proliferator-activated receptor gamma1 (PPAR-gamma1) as a major PPAR in a tissue in which estrogen induces peroxisome proliferation. AB - Estradiol administration induces peroxisome proliferation and the production of 3 hydroxy fatty acid pheromones in the uropygial glands of the duck, but not in the goose gland, which does not produce such pheromones. We isolated a peroxisome proliferator-activated receptor (PPAR)gamma1 cDNA from a duck uropygial gland cDNA library. Northern blots revealed two transcripts, PPAR gamma1 and gamma2, and showed that PPAR gamma was expressed at higher levels than PPAR alpha in the uropygial gland of the duck. Although PPAR gamma2 was expressed in both duck and goose uropygial gland, PPAR gamma1 was expressed only in the duck gland, which responds to estrogen by peroxisome proliferation. In NIH 3T3 transfected cells, PPAR gamma1 was activated by peroxisome proliferators such as Wy-14643, clofibric acid and Ly-171883 causing induction of the target marker gene. By cotransfection with a plasmid containing alpha-cis-retinoic acid receptor RXR alpha, the induction increased up to 9-fold. These results suggest that PPAR gamma1 may be involved in peroxisome proliferation while PPAR gamma2 may be involved in lipid metabolism. PMID- 9742962 TI - The proregion of papaya proteinase IV inhibits Colorado potato beetle digestive cysteine proteinases. AB - Three distinct digestive protease systems were induced in larvae of the herbivorous pest, Colorado potato beetle (CPB; Leptinotarsa decemlineata Say), and used as a model to assess the ability of the proregion of papaya proteinase IV (PPIV; glycyl endopeptidase, EC 3.4.22.25) to act as an inhibitor of insect digestive cysteine proteinases. As shown by gelatin/PAGE and complementary inhibition assays, a recombinant form of the proregion produced in Escherichia coli inhibited a fraction of the insect proteases also inhibited by the well characterized inhibitor of cysteine proteinases, oryzacystatin I (OCI). In contrast with OCI, the inhibitory potency of the proregion was affected by an increase of the temperature, suggesting a certain alteration of its structural integrity by the insect non-target proteases. This apparent susceptibility to proteolysis was confirmed by SDS-PAGE, after challenging the proregion with the different insect extracts. As seen on gel, selective inhibition of the insect aspartate proteinase, cathepsin D, with the inhibitor pepstatin A preserved the activity of the proregion against cysteine proteinases by preventing its hydrolysis. Taken together, these observations suggest the potential of plant protease proregions as regulators of cysteine proteinases in biotechnological systems, and show the ability of protease inhibitors to preserve the integrity of 'companion' defense-related proteins from the action of insensitive proteases in target pests. PMID- 9742963 TI - Effect of squalene synthase gene disruption on synthesis of polyprenols in Saccharomyces cerevisiae. AB - Biosynthesis of polyprenols was investigated in a wild-type strain of Saccharomyces cerevisiae and a squalene synthase deficient strain auxotrophic for ergosterol. The quantitative data showed that disruption of squalene synthase gene caused a 6-fold increase in the synthesis of polyprenols in vitro in comparison with the wild-type strain. Microsomal preparation from the deleted strain only slightly reacted to the additional exogenous FPP, while that from the wild-type strain presented a 4-fold increase of polyprenol synthesis. Restoration of ergosterol synthesis, by introducing ERG9 functional allele into the deleted strain resulted in a significant lowering of polyprenol synthesis, indicating the immediate shift of the common substrate (FPP) to the sterol pathway. The role of squalene synthase in the regulation of polyprenol synthesis and 'flow diversion hypothesis' is discussed. PMID- 9742964 TI - Telomerase activity in 'immortal' fish. AB - Eukaryotic chromosome termini consist of telomeres, short sequence repeats. According to the telomere hypothesis, DNA replication leads to telomere shortening, resulting in a cellular mitotic clock. Telomerase resets it by telomere synthesis. In mammals with a limited growth phase, telomerase activity in somatic tissues is restricted to stem cell derivatives with high proliferation potential. But other animals, like some fish, grow throughout their life with little senescence. All somatic cells require a high proliferation capacity and telomerase should be active in all cells, irrespective of fish age. Indeed, we detected high telomerase activities in all analyzed organs of rainbow trout (Oncorhynchus mykiss). PMID- 9742965 TI - The iridoid glucoside secologanin is derived from the novel triose phosphate/pyruvate pathway in a Catharanthus roseus cell culture. AB - Secologanin is the iridoid building block of the majority of the terpenoid indole alkaloids. In the biosynthesis of secologanin, mevalonate was considered to be the exclusive precursor of isopentenyl diphosphate. After [1-(13)C]glucose feeding to a cell culture of Catharanthus roseus, its incorporation into secologanin was studied by 13C NMR spectroscopy. The data showed that the novel triose phosphate/pyruvate and not the mevalonate pathway was the major route for the biosynthesis of secologanin. PMID- 9742966 TI - The vascular endothelial growth factor mRNA contains an internal ribosome entry site. AB - Vascular endothelial growth factor (VEGF), an essential regulator of angiogenesis during early development as well as during the growth of solid tumours, bears an unusually large 5' untranslated region (5'-UTR) in the mRNA of over 1000 nucleotides. We found that the VEGF 5'-UTR, despite being GC-rich and containing an upstream short open reading frame, promotes efficient translation of a luciferase reporter. The VEGF 5'-UTR also allowed translation of luciferase from a dicistronic mRNA when placed between the two cistrons, demonstrating that it contains an internal ribosome entry site. Deletion analysis indicated that the IRES resides towards the 3' end of the 5'-UTR. PMID- 9742967 TI - Activation of nitric oxide synthase is involved in tamoxifen-induced apoptosis of human erythroleukemia K562 cells. AB - Tamoxifen induces apoptosis (programmed cell death) in human erythroleukemia K562 cells. Nitric oxide synthase activity and expression increased in apoptotic cells by 315% and 280%, respectively, compared to controls. The specific inhibitor of nitric oxide synthase, L-NAME, protected K562 cells from tamoxifen-induced apoptosis, whereas the nitric oxide donor, sodium nitroprusside (SNP), potentiated the apoptotic effect of the drug. In addition, 5-lipoxygenase was activated by tamoxifen and the specific enzyme inhibitor, MK886, protected K562 cells against the drug. Conversely, the 5-lipoxygenase product, 5 hydroperoxyeicosatetraenoic acid, enhanced the tamoxifen-induced apoptosis. Finally, tamoxifen altered also membrane properties of K562 cells. PMID- 9742968 TI - Targeting of thylakoid proteins by the delta pH-driven twin-arginine translocation pathway requires a specific signal in the hydrophobic domain in conjunction with the twin-arginine motif. AB - Superficially similar cleavable targeting signals specify whether lumenal proteins are transported across the thylakoid membrane by a Sec- or delta pH dependent pathway. A twin-arginine motif is essential but not sufficient to direct delta pH-dependent targeting; here we show that a second determinant is located in the hydrophobic region. A highly hydrophobic amino acid is found either two or three residues C-terminal to the twin-arginine in all known transfer peptides for the delta pH-dependent system, and substitution of this residue in the 23-kDa (23K) peptide markedly inhibits translocation. Further, whereas the insertion of twin-arginine in a Sec-dependent precursor does not permit efficient delta pH-dependent targeting, the simultaneous presence of a leucine at the +3 position (relative to the RR) enables the peptide to function as efficiently as an authentic transfer peptide. RRNVL, RRAAL and RRALA within a Sec targeting signal all support efficient delta pH-dependent targeting, RRNVA is less effective and RRNAA/RRNAG are totally ineffective. We conclude that the core signal for this pathway is a twin-arginine together with an adjacent hydrophobic determinant. PMID- 9742969 TI - Tyrosine phosphorylation of the Wiskott-Aldrich syndrome protein by Lyn and Btk is regulated by CDC42. AB - The Wiskott-Aldrich syndrome (WAS) is a rare immunodeficiency disease affecting mainly platelets and lymphocytes. Here, we show that the WAS gene product, WASp, is tyrosine phosphorylated upon aggregation of the high affinity IgE receptor (Fc epsilonRI) at the surface of RBL-2H3 rat tumor mast cells. Lyn and the Bruton's tyrosine kinase (Btk), two protein tyrosine kinases involved in Fc epsilonRI signaling phosphorylate WASp and interact with WASp in vivo. Interestingly, expression of a GTPase defective mutant form of CDC42, that interacts with WASp, is accompanied by a substantial increase in WASp tyrosine phosphorylation. This study suggests that activated CDC42 recruits WASp to the plasma membrane where it becomes phosphorylated by Lyn and Btk. We conclude that WASp represents a connection between protein tyrosine kinase signaling pathways and CDC42 function in cytoskeleton and cell growth regulation in hematopoietic cells. PMID- 9742970 TI - 15-Lipoxygenation of phospholipids may precede the sn-2 cleavage by phospholipases A2: reaction specificities of secretory and cytosolic phospholipases A2 towards native and 15-lipoxygenated arachidonoyl phospholipids. AB - Reticulocyte-type 15-lipoxygenase is known to dioxygenate phospholipids without preceding action of phospholipases A2 (PLA2). Therefore we studied the reaction of the secretory PLA2s (sPLA2) from pancreas and snake venom, and of the human cytosolic PLA2 (cPLA2) with 1-palmitoyl-2-arachidonoyl phosphatidylcholine (PAPC) and their 15-lipoxygenated species (PAPC-OOH and PAPC-OH) either alone or as equimolar mixtures. These PLA2s cleaved PAPC-O(O)H with higher (sPLA2) or similar rates (cPLA2) as compared with native PAPC. In mixtures, however, PAPC proved to be the preferred, albeit not exclusive substrate for all three PLA2s. Thus, partial 15-lipoxygenation of phospholipids may also trigger liberation of arachidonic acid. PMID- 9742971 TI - Paying for family planning. PMID- 9742972 TI - Some answers, more controversy, from UKPDS. United Kingdom Prospective Diabetes Study. PMID- 9742973 TI - Radiation therapy for benign disease: an old area revisited. PMID- 9742974 TI - Gains and losses from dexamethasone for neonatal chronic lung disease. PMID- 9742975 TI - Open debate on hormone-replacement therapy. PMID- 9742976 TI - Intensive blood-glucose control with sulphonylureas or insulin compared with conventional treatment and risk of complications in patients with type 2 diabetes (UKPDS 33). UK Prospective Diabetes Study (UKPDS) Group. AB - BACKGROUND: Improved blood-glucose control decreases the progression of diabetic microvascular disease, but the effect on macrovascular complications is unknown. There is concern that sulphonylureas may increase cardiovascular mortality in patients with type 2 diabetes and that high insulin concentrations may enhance atheroma formation. We compared the effects of intensive blood-glucose control with either sulphonylurea or insulin and conventional treatment on the risk of microvascular and macrovascular complications in patients with type 2 diabetes in a randomised controlled trial. METHODS: 3867 newly diagnosed patients with type 2 diabetes, median age 54 years (IQR 48-60 years), who after 3 months' diet treatment had a mean of two fasting plasma glucose (FPG) concentrations of 6.1 15.0 mmol/L were randomly assigned intensive policy with a sulphonylurea (chlorpropamide, glibenclamide, or glipizide) or with insulin, or conventional policy with diet. The aim in the intensive group was FPG less than 6 mmol/L. In the conventional group, the aim was the best achievable FPG with diet alone; drugs were added only if there were hyperglycaemic symptoms or FPG greater than 15 mmol/L. Three aggregate endpoints were used to assess differences between conventional and intensive treatment: any diabetes-related endpoint (sudden death, death from hyperglycaemia or hypoglycaemia, fatal or non-fatal myocardial infarction, angina, heart failure, stroke, renal failure, amputation [of at least one digit], vitreous haemorrhage, retinopathy requiring photocoagulation, blindness in one eye, or cataract extraction); diabetes-related death (death from myocardial infarction, stroke, peripheral vascular disease, renal disease, hyperglycaemia or hypoglycaemia, and sudden death); all-cause mortality. Single clinical endpoints and surrogate subclinical endpoints were also assessed. All analyses were by intention to treat and frequency of hypoglycaemia was also analysed by actual therapy. FINDINGS: Over 10 years, haemoglobin A1c (HbA1c) was 7.0% (6.2-8.2) in the intensive group compared with 7.9% (6.9-8.8) in the conventional group--an 11% reduction. There was no difference in HbA1c among agents in the intensive group. Compared with the conventional group, the risk in the intensive group was 12% lower (95% CI 1-21, p=0.029) for any diabetes-related endpoint; 10% lower (-11 to 27, p=0.34) for any diabetes-related death; and 6% lower (-10 to 20, p=0.44) for all-cause mortality. Most of the risk reduction in the any diabetes-related aggregate endpoint was due to a 25% risk reduction (7 40, p=0.0099) in microvascular endpoints, including the need for retinal photocoagulation. There was no difference for any of the three aggregate endpoints between the three intensive agents (chlorpropamide, glibenclamide, or insulin). Patients in the intensive group had more hypoglycaemic episodes than those in the conventional group on both types of analysis (both p<0.0001). The rates of major hypoglycaemic episodes per year were 0.7% with conventional treatment, 1.0% with chlorpropamide, 1.4% with glibenclamide, and 1.8% with insulin. Weight gain was significantly higher in the intensive group (mean 2.9 kg) than in the conventional group (p<0.001), and patients assigned insulin had a greater gain in weight (4.0 kg) than those assigned chlorpropamide (2.6 kg) or glibenclamide (1.7 kg). INTERPRETATION: Intensive blood-glucose control by either sulphonylureas or insulin substantially decreases the risk of microvascular complications, but not macrovascular disease, in patients with type 2 diabetes.(ABSTRACT TRUNCATED) PMID- 9742977 TI - Effect of intensive blood-glucose control with metformin on complications in overweight patients with type 2 diabetes (UKPDS 34). UK Prospective Diabetes Study (UKPDS) Group. AB - BACKGROUND: In patients with type 2 diabetes, intensive blood-glucose control with insulin or sulphonylurea therapy decreases progression of microvascular disease and may also reduce the risk of heart attacks. This study investigated whether intensive glucose control with metformin has any specific advantage or disadvantage. METHODS: Of 4075 patients recruited to UKPDS in 15 centres, 1704 overweight (>120% ideal bodyweight) patients with newly diagnosed type 2 diabetes, mean age 53 years, had raised fasting plasma glucose (FPG; 6.1-15.0 mmol/L) without hyperglycaemic symptoms after 3 months' initial diet. 753 were included in a randomised controlled trial, median duration 10.7 years, of conventional policy, primarily with diet alone (n=411) versus intensive blood glucose control policy with metformin, aiming for FPG below 6 mmol/L (n=342). A secondary analysis compared the 342 patients allocated metformin with 951 overweight patients allocated intensive blood-glucose control with chlorpropamide (n=265), glibenclamide (n=277), or insulin (n=409). The primary outcome measures were aggregates of any diabetes-related clinical endpoint, diabetes-related death, and all-cause mortality. In a supplementary randomised controlled trial, 537 non-overweight and overweight patients, mean age 59 years, who were already on maximum sulphonylurea therapy but had raised FPG (6.1-15.0 mmol/L) were allocated continuing sulphonylurea therapy alone (n=269) or addition of metformin (n=268). FINDINGS: Median glycated haemoglobin (HbA1c) was 7.4% in the metformin group compared with 8.0% in the conventional group. Patients allocated metformin, compared with the conventional group, had risk reductions of 32% (95% CI 13-47, p=0.002) for any diabetes-related endpoint, 42% for diabetes-related death (9-63, p=0.017), and 36% for all-cause mortality (9-55, p=0.011). Among patients allocated intensive blood-glucose control, metformin showed a greater effect than chlorpropamide, glibenclamide, or insulin for any diabetes-related endpoint (p=0.0034), all-cause mortality (p=0.021), and stroke (p=0.032). Early addition of metformin in sulphonylurea-treated patients was associated with an increased risk of diabetes-related death (96% increased risk [95% CI 2-275], p=0.039) compared with continued sulphonylurea alone. A combined analysis of the main and supplementary studies showed fewer metformin-allocated patients having diabetes related endpoints (risk reduction 19% [2-33], p=0.033). Epidemiological assessment of the possible association of death from diabetes-related causes with the concurrent therapy of diabetes in 4416 patients did not show an increased risk in diabetes-related death in patients treated with a combination of sulphonylurea and metformin (risk reduction 5% [-33 to 32], p=0.78). INTERPRETATION: Since intensive glucose control with metformin appears to decrease the risk of diabetes-related endpoints in overweight diabetic patients, and is associated with less weight gain and fewer hypoglycaemic attacks than are insulin and sulphonylureas, it may be the first-line pharmacological therapy of choice in these patients. PMID- 9742978 TI - CCR5 promoter polymorphism and HIV-1 disease progression. Multicenter AIDS Cohort Study (MACS). AB - BACKGROUND: The rate of progression to AIDS varies among individuals infected with HIV-1. Factors responsible include two inherited human alleles, CCR5 delta32 and CCR2-641, which alter the protein-coding regions for the HIV-1 coreceptors/chemokine receptors CCR5 and CCR2b. We tested the hypothesis that polymorphisms of the CCR5 promoter might affect the rate of progression of HIV-1 infected people to AIDS. METHODS: We used directed heteroduplex analysis to identify polymorphism in the CCR5 promoter. Promoter-variants were compared in vitro with a chloramphenicol acetyltransferase reporter gene, and in vivo by genotyping HIV-1 seroconvertors discordant at polymorphous loci. FINDINGS: An A/G polymorphism was identified at basepair 59029 (Genbank U95626) in the CCR5 promoter. Both promoter alleles were common (43-68% allelic frequency for 59029-A depending on race). When in-vitro promoter activity was measured, 59029-G had 45% lower activity than 59029-A (p=0.05). In a cohort of HIV-1 seroconvertors lacking both CCR5 delta32 and CCR2-641, 59029-G/G individuals progressed to AIDS on average 3.8 years more slowly than 59029-A/A individuals (p=0.004). 59029-G/A discordance did not correlate with discordant rates of infection. INTERPRETATION: Our results are consistent with the hypothesis that CCR5 is important in HIV-1 pathogenesis. CCR5 59029-G/G appears to be protective relative to CCR5 59029-A/A, and about twice as protective relative to CCR5 delta32 or CCR2-641. This effect may be the result of reduced CCR5 mRNA production. These results identify the first site in the CCR5 promoter that may be a useful target for treatment of HIV 1 infection. PMID- 9742979 TI - p53 codon 72 polymorphism and risk of cervical cancer in UK. AB - BACKGROUND: A polymorphism at codon 72 of the human tumour-suppressor gene, p53, results in translation to either arginine or proline. A recent report suggested that the risk of human-papillomavirus-associated cervical cancer in white women is higher for those homozygous for the arginine allele than for those who are heterozygous. We examined a similar number of cervical cancers and a larger control group for their p53 codon 72 polymorphism status to see if we could confirm this result. METHODS: Three different groups of UK white women were studied: 96 who had volunteered to take part in a trial of ovarian-cancer screening; 150 attending for routine antenatal care in the Oxford region; and 50 women with cervical cancer. DNA from peripheral blood samples and from archival tissue samples was examined by PCR with allele-specific primers. FINDINGS: The proportions of individuals homozygous for the arginine allele, homozygous for the proline allele, and heterozygous for the two alleles were 59%, 4%, and 36% among women screened for ovarian cancer; 65%, 8%, and 27% among the antenatal-care group; and 54%, 6%, and 40% in women with cervical cancer. Chi2 analysis showed no significant differences in these proportions. INTERPRETATION: In the population studied, individuals homozygous for the arginine variant of codon 72 of the p53 gene were not at increased risk of cervical cancer. PMID- 9742980 TI - PCR on cerebrospinal fluid to show influenza-associated acute encephalopathy or encephalitis. AB - BACKGROUND: Except for Reye's syndrome, influenza-associated acute encephalopathy or encephalitis is not universally recognised. We did a multicentre study of laboratory and clinical data for patients with influenza-associated acute encephalopathy or encephalitis. METHODS: In Nagoya, Japan, ten patients with acute encephalopathy or encephalitis associated with influenza-like illness were admitted to our hospitals between April, 1996, and March, 1997. We collected clinical, laboratory and serological data and assessed cerebrospinal fluid samples by PCR for influenza A and B. FINDINGS: Seven patients, aged 22 months to 4 years, had evidence of recent influenza infection, six with type-A/Hong Kong (H3N2) and one with type B. The first sign in the central nervous system appeared within 2 days of fever in all but one patient. The first sign of involvement of the central nervous system was generalised convulsions in all patients. Two patients died, one had sequelae, and four survived without sequelae. PCR for influenza type A was positive for five patients. INTERPRETATION: The results of PCR suggest that at least part of the influenza type A genome existed in the central nervous system. Influenza-associated acute encephalopathy or encephalitis in young children deserves wider recognition. PMID- 9742981 TI - Intestinal strictures. PMID- 9742982 TI - U-shaped relation for alcohol consumption and health in early adulthood and implications for mortality. PMID- 9742983 TI - Regression of autoimmune thrombocytopenia after eradication of Helicobacter pylori. PMID- 9742984 TI - Severe Raynaud's phenomenon associated with interferon-beta treatment for multiple sclerosis. PMID- 9742985 TI - Hepatocytes as a source of collagen type XVIII endostatin. PMID- 9742986 TI - Risk of aspirin use plus thrombolysis after acute ischaemic stroke: a further MAST-I analysis. MAST-I Collaborative Group. Multicentre Acute Stroke Trial- Italy. PMID- 9742987 TI - Remission and immune reconstitution after T-cell-depleted stem-cell transplantation for rheumatoid arthritis. PMID- 9742988 TI - Decreased neuronal inhibition in cerebral cortex in obsessive-compulsive disorder on transcranial magnetic stimulation. PMID- 9742989 TI - Suicide with "non-lethal" firearm. PMID- 9742990 TI - Tamoxifen prevention claim will not be allowed in USA. PMID- 9742991 TI - Hantaviruses: an emerging threat to human health? PMID- 9742992 TI - Investigating the reasons for Spain's falling birth rate. PMID- 9742993 TI - Failure to inform sex partner of HIV status assault in Canada. PMID- 9742994 TI - South African government urged to show more in tackling AIDS. PMID- 9742995 TI - Publication and promotion. A fair reward. PMID- 9742996 TI - Publication and promotion. The performance of science. PMID- 9742997 TI - Publication and promotion. The public interest. PMID- 9742998 TI - Publication and promotion. Is the system really broken? PMID- 9742999 TI - Publication and promotion. Intelligence work. PMID- 9743000 TI - Publication and promotion. The value of discrimination. PMID- 9743001 TI - Publication and promotion. Drafter and draftees. PMID- 9743003 TI - Publication and promotion. Open your eyes. PMID- 9743002 TI - Publication and promotion. Writing is all. PMID- 9743004 TI - Publication and promotion. Long live the deans! PMID- 9743005 TI - Treatment of systemic lupus erythematosus: from cod-liver oil to cyclosporin. PMID- 9743006 TI - The new new public health. PMID- 9743007 TI - The new new public health. PMID- 9743008 TI - The new new public health. PMID- 9743009 TI - Use of Pro-Gest cream in postmenopausal women. PMID- 9743010 TI - Use of Pro-Gest cream in postmenopausal women. PMID- 9743011 TI - Use of Pro-Gest cream in postmenopausal women. PMID- 9743012 TI - Spectrum of AIDS-associated malignant disorders. PMID- 9743013 TI - Spectrum of AIDS-associated malignant disorders. PMID- 9743014 TI - Predictions of liver histology from laboratory tests and demographic data [corrected]. PMID- 9743015 TI - Overlooking orthostatic hypotension. PMID- 9743016 TI - Combined interferon-alpha and interleukin-2 as adjuvant treatment for melanoma. PMID- 9743017 TI - Statins for prevention of stroke. PMID- 9743018 TI - Statins for prevention of stroke. PMID- 9743019 TI - Does biopsy before resection compromise prognosis in colorectal cancer? PMID- 9743020 TI - Siblings of centenarians live longer: a computer simulation. PMID- 9743021 TI - Trauma, post-traumatic stress disorder, and war. PMID- 9743022 TI - Obesity: a time bomb to be defused. PMID- 9743023 TI - The Doctor. PMID- 9743024 TI - An artist in hospital: a sketchbook of the Dutch impressionist Isaac Israels. PMID- 9743025 TI - Interactive pregnancy, parenting, and ob/gyn websites. PMID- 9743026 TI - The role of the primary pediatrician when a child dies. PMID- 9743027 TI - National Institutes of Health support for research and training: future of pediatrician scientists. PMID- 9743028 TI - Variation among neonatal intensive care units in narcotic administration. AB - OBJECTIVES: To compare rates of narcotic administration for medically treated neonates in different neonatal intensive care units (NICUs) and to compare treated and untreated neonates to assess whether narcotics provided advantages or disadvantages for short-term outcomes, such as cardiovascular stability (ie, blood pressure and heart rate), hyperbilirubinemia, duration of respiratory support, growth, and the incidence of intraventricular hemorrhage. STUDY DESIGN: The medical charts of neonates weighing less than 1500 g, admitted to 6 NICUs (A F), were abstracted. Neonates who had a chest tube or who had undergone surgery were excluded from the study, leaving the records of 1171 neonates. We modeled outcomes by linear or logistic regression, controlling for birth weight (<750, 750-999, and 1000-1499 g) and illness severity (low, 0-9; medium, 10-19; high, > or =20) using the Score for Neonatal Acute Physiology (SNAP), and adjusted for NICU. RESULTS: Narcotic use varied by birth weight (<750 g, 21%; 750-999 g, 13%; and 1000-1499 g, 8%), illness severity (low, 9%; medium, 19%; and high, 37%), day (1, 11%; 3, 6%; and 14, 2%), and NICU. We restricted analyses to the 1018 neonates who received mechanical ventilation on day 1. Logistic regression, adjusting for birth weight and SNAP, confirmed a 28.6-fold variation in narcotic administration (odds ratios, 4.1-28.6 vs NICU A). Several short-term outcomes also were associated with narcotic use, including more than 33 g of fluid retention on day 3 and a higher direct bilirubin level (6.8 micromol/L higher [0.4 mg/dL higher], P = .03). There were no differences in weight gain at 14 and 28 days or mechanical ventilatory support on days 14 and 28. Narcotic use was not associated with differences in worst blood pressure or heart rate or with increased length of hospital stay. CONCLUSIONS: Our study found a 28.6-fold variation among NICUs in narcotic administration in very low-birth-weight neonates. We were unable to detect any major advantages or disadvantages of narcotic use. We did not assess iatrogenic abstinence syndrome or long-term outcomes. These results indicate the need for randomized trials to rationalize these widely differing practices. PMID- 9743029 TI - Pediatric death certification. AB - OBJECTIVE: To determine location, manner, and physician certifier of pediatric deaths. DESIGN: A descriptive study of death certificate information for all child deaths (aged birth through 17 years) for the years 1995 and 1996. SETTING: Urban county of more than 780,000 population. MAIN OUTCOME MEASURES: Field of specialty of physician certifiers, location of death, and category of deaths certified by the medical examiner. RESULTS: Of 361 child deaths, 42.6% were certified by the medical examiner, 24.1% by neonatologists, 10.0% by obstetricians, 8.0% by pediatric critical care specialists, and 5.3% by general pediatricians. The remaining deaths were certified by pediatric subspecialists, surgeons, family practitioners, emergency medicine specialists, hospital pathologists, and law enforcement officials. The medical examiner certified deaths due to trauma (64.5%), sudden infant death syndrome (13.5%), unexplained or suspicious causes (9.7%), medical or surgical complications (3.9%), or because no other physician certifier was available (5.8%). Most children were pronounced dead at hospitals, but 10.0% died at home, 4.4% on roads, and 2.5% on public or private lands. CONCLUSIONS: General pediatricians are unlikely to be directly involved in the care of most children who die and are therefore unlikely to sign the death certificate. Education about death and dying issues should be available for all pediatricians but should be directed at those specialists most likely to provide care during critical events. Support services for families need to be community based and accessible to survivors. PMID- 9743031 TI - Strategies to promote breast-feeding among adolescent mothers. AB - OBJECTIVE: To identify characteristics of adolescent mothers who bottle-feed who considered breast-feeding their infants and strategies to promote breast-feeding within this special group. DESIGN: Adolescents completed an hour-long interview within 48 hours of delivery that elicited factors considered important to the mother's feeding decision and indices of mental health. SETTING: Postpartum ward of university hospital. SUBJECTS: A total of 693 adolescents 18 years old or younger (mean age, 16.7 years) from African American, Mexican American, or white race or ethnicity; 27% of Mexican American participants spoke little or no English. MAIN OUTCOME MEASURES: Factors associated with breast-feeding decision. RESULTS: Those who chose bottle-feeding (hereafter, bottle-feeders) who had considered breast-feeding were first compared with bottle-feeders who had not considered breast-feeding and then with adolescents who breast-fed. After controlling for ethnicity, bottle-feeders who had considered breast-feeding were more likely than those who had not considered breast-feeding to be impoverished (adjusted odds ratio [AOR] = 4.8), to have delayed their feeding decision until the later stages of pregnancy (AOR = 4.6), to have been encouraged to breast-feed (AOR = 4.5), to have friends who breast-fed (AOR = 2.3), and to have experienced low financial, tangible, emotional, or informational support from their families (AOR = 1.6). They were more likely to cite barriers associated with breast feeding while returning to school or work (AOR = 2.0) and less likely to state that bottle-feeding was healthier (AOR = 0.3) as reasons for bottle-feeding. Compared with those who chose breast-feeding (hereafter, breast-feeders), this group was more likely to have made the feeding decision alone rather than relying on advice (AOR = 4.6), to have made this decision in the later stages of pregnancy (AOR = 4.4), to report fewer breast-feeding role models (AOR = 1.8) and fewer significant others who encouraged breast-feeding (AOR = 2.8), and to report at least 2 significant others who encouraged bottle-feeding (AOR= 3.2). They were also less likely to have attempted to breast-feed a previous child (AOR = 3.3). CONCLUSIONS: A subgroup of adolescent mothers who had considered breast-feeding but ultimately chose to bottle-feed may be identified in the late stages of gestation by collecting information on financial status, family support, perceived barriers to breast-feeding and attending school or working, timing of the feeding decision, prior breast-feeding experience, breast-feeding role models, and encouragement to breast-feed. We speculate that strategies to promote breast-feeding should focus on role modeling and facilitation. PMID- 9743030 TI - Predictors of child psychological changes during family-based treatment for obesity. AB - OBJECTIVE: To explore the influence of 1-year changes in child obesity and maternal psychopathology on changes in child psychological problems. DESIGN: Hierarchical regression models were used to predict child psychological change, with demographic variables, maternal psychological change, and child percentage overweight change as predictors. SETTING: Pediatric obesity research clinic. PARTICIPANTS: Clinic sample of 116 obese 8- to 12-year-old children and their mothers. INTERVENTIONS: Family-based behavioral weight-control program. MAIN OUTCOME MEASURES: Child psychopathology was assessed via mother-reported Child Behavior Checklists and maternal psychopathology was determined by standardizing scores on the Cornell Medical Index and the Symptoms Checklist-90-Revised. RESULTS: Significant improvements were observed in child percentage overweight ( 20.1% overweight), and child and maternal psychopathology. Improved maternal psychopathology accounted for a significant amount of variance in improvements in the Child Behavior Checklist total Problems Scale and internalizing and externalizing problems subscales. Decreased obesity accounted for a significant amount of variance in improvements in the Total Competence scale and, somatic complaints, social problems and social competence subscales of the Child Behavior Checklist. Significant interactions of child obesity change by sex were found for Total Problems and externalizing scores. The interactions were due to girls with greater obesity reduction showing greater improvement in Total Problems, whereas boys with greater obesity reduction showed less improvement in externalizing problems. CONCLUSIONS: These results highlight the multidimensional nature of psychosocial functioning in obese children and call attention to multiple avenues for intervention to improve their psychosocial functioning. PMID- 9743032 TI - Disordered eating among adolescents with chronic illness and disability: the role of family and other social factors. AB - OBJECTIVES: To compare prevalence rates of weight-control behaviors among adolescents with and without chronic illness and to explore the role of familial and other social factors on associations between disordered eating and chronic illness. DESIGN AND SETTING: Survey conducted in public schools in Connecticut. PARTICIPANTS: A representative statewide population-based sample of 9343 7th-, 9th-, and 11th-grade public school students, of whom 1021 reported a chronic illness. MAIN OUTCOME MEASURES: Disordered eating (vomiting, diet pills, and laxatives), dieting, and exercise for weight control; chronic illness status; family structure, family communication, parental caring, parental monitoring, parental expectations, peer support, and sexual and physical abuse. RESULTS: Adolescents with chronic illness were at greater risk for disordered eating than youth without chronic illness, after controlling for sociodemographic variables (girls: odds ratio, 1.59 [95% confidence interval, 1.19-2.14]; boys: odds ratio, 2.22 [95% confidence interval, 1.49-3.32]). Adolescents with chronic illness were less likely to come from 2-parent families; reported lower levels of family communication, parental caring, and parental expectations; and reported more sexual and physical abuse than youth without chronic illness. Male adolescents with chronic illness were more likely to report low peer support and low parental monitoring. Most of these familial-social factors were also associated with an increased prevalence of disordered eating. After familial-social factors were controlled for, however, associations between disordered eating and chronic illness remained statistically significant. CONCLUSIONS: Adolescents with chronic illness are at greater risk for disordered eating behaviors than youth without chronic illness. Factors other than the familial-social factors assessed in this study may be contributing to this increased risk. In the clinical setting, youth with chronic illness need to be screened for disordered eating and familial and other social concerns. PMID- 9743033 TI - Disappearance of vesicoureteral reflux in children. AB - OBJECTIVE: To describe the disappearance of reflux in children with vesicoureteral reflux, in whom there are presently no population-based long-term studies. DESIGN: An unselected cohort of children with reflux detected after their first known symptomatic urinary tract infection was followed up prospectively for up to 15 years. SETTING: A single children's hospital in a distinct geographical area at which most children with symptomatic urinary tract infection were treated. PATIENTS: Two hundred thirty children--173 girls and 57 boys--with unilateral (n=130) and bilateral (n=100) reflux. Dilated reflux (grades III-V) was found in 54 patients (23.5%). The frequency of reflux was 34% in girls and 31% in boys who were examined after urinary tract infection. MAIN OUTCOME MEASURE: Disappearance of reflux. RESULTS: The probability of spontaneous disappearance of reflux was estimated using Kaplan-Meier survival curves based on 164 children who underwent multiple voiding cystourethrographies. There was a marked tendency for disappearance of reflux, with 73% of children with dilated reflux having no or only grade I reflux after 10 years. Shorter persistence of reflux was found in children with undilated reflux at the initial investigation and in boys compared with girls. However, age at first investigation was not related to the rate of disappearance, and there was no difference between children with bilateral compared with unilateral reflux. CONCLUSIONS: This study of an unselected group of children with urinary tract infection shows a favorable long-term outcome concerning disappearance of reflux. In children with dilated reflux, this tendency was more pronounced than previously reported. PMID- 9743034 TI - Problem dieting behaviors among young adolescents. AB - OBJECTIVE: To examine dieting, eating and exercise behaviors, use of diet pills, and vomiting or use of laxatives to lose weight among younger adolescents. DESIGN: Analysis of data from a modified version of the Youth Risk Behavior Survey administered to middle school students in North Carolina in 1995. SETTING: Fifty-three randomly selected middle schools in North Carolina. SUBJECTS: Two thousand three hundred thirty-one students in the sixth, seventh, and eighth grades. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Responses to questions regarding weight control practices, including vomiting or laxative use, dieting, exercise, or diet pill use. RESULTS: Of the students surveyed, 110 (9.7%) of the girls and 46 (4.0%) of the boys reported vomiting or using laxatives to lose weight. Among the girls, vomiting or laxative use was associated with feeling overweight, other weight loss practices, older age, being a poor student, smoking, eating more salads or vegetables, and eating more candy or other sweets (P< or =.01). A logistic regression model consisting of diet pill use, dieting to lose weight, lower academic achievement, and currently trying to lose weight correctly classified 92% of female students who had or had not vomited or used laxatives. Among boys, vomiting or laxative use was associated with feeling overweight, other weight loss practices, minority racial status, smoking, frequency of eating hamburgers or other high-fat meats, and frequency of eating french fries or potato chips (P< or =.01). A model consisting of diet pill use, minority race, dieting to lose weight, smoking, feeling overweight, and number of servings of hamburgers, hot dogs, or barbecue correctly classified 97% of the boys who had or had not vomited or used laxatives. CONCLUSION: Younger adolescents trying to lose weight engage in a variety of problem dieting and weight loss behaviors that can compromise health and may be associated with eating disorders. PMID- 9743035 TI - Parental barriers to weaning infants from the bottle. AB - BACKGROUND: Optimal bottle weaning should occur between 12 and 15 months of age. We hypothesized that high-risk populations have different parental attitudes, learned behaviors, and knowledge of weaning practices. OBJECTIVE: To determine whether high-risk populations are less likely to wean their children by 15 months of age than low-risk populations. METHODS: A cross-sectional survey using a convenience sample of parents was conducted at 3 community-based pediatric clinics. Spanish- and English-speaking parents with weaned and unweaned children 12 to 36 months of age were included in the study. A self-administered questionnaire was completed at a clinic visit. The questionnaire addressed aspects of parents' sociodemographic characteristics and included feeding history; weaning practices; sources of information about weaning; and parental behaviors, attitudes, and knowledge of age at which the child should be weaned. RESULTS: One hundred eighty questionnaires were completed. Marital status was related to weaning behavior. Seventy-six percent of single mothers had weaned their children in a timely manner, whereas 48% of married mothers had done so (chi2 = 7.70; P = .008). Parental education, race, and income were not significantly related to the timeliness of weaning. When respondents rated the helpfulness of multiple sources, only the health clinic was found to be significantly more important for the timely weaning group (t = -2.13; P = .04). Parents with timely weaned children stated that the mean +/- SD optimal age for weaning is 13.6 +/- 3.2 months. Parents with unweaned and late-weaned children stated that the mean +/- SD optimal age is 19.9 +/- 6.6 months. Bedtime bottle feedings were reported in more than 87% of the unweaned group. Sixty-nine percent reported poor dental development associated with delayed weaning. CONCLUSIONS: Married parents are at risk of late weaning. Parents continue to allow their children to sleep with milk bottles in their mouths in bed at night. Parents are not aware of the medical problems associated with late weaning. Late-weaning parents are not knowledgeable about current weaning recommendations. Current approaches are not effective in altering set patterns of inappropriate weaning habits. Additional interventions and innovative parental education methods are needed to improve age-appropriate weaning practices. PMID- 9743036 TI - Correlates and consequences of early removal of levonorgestrel implants among teenaged mothers. AB - OBJECTIVE: To determine if adolescent mothers who request early removal of levonorgestrel implants differ from those who do not in ways that might predispose them to repeated conceptions and in their concerns about adverse effects. We hypothesized that adolescent mothers who request removal of levonorgestrel implants within 2 years of insertion have more risk factors for repeated pregnancy than those who do not. METHODS: We studied the prevalence of 21 characteristics associated with repeated adolescent pregnancy and 16 adverse effects of levonorgestrel implants in 181 postpartum, adolescent levonorgestrel implant recipients, 66 (36%) of whom had the levonorgestrel implants removed within 20 months of insertion (hereafter, removers). RESULTS: Removers (n = 66) had significantly more risk factors for repeated pregnancy and reported significantly more adverse effects than did those who continued to use levonorgestrel implants (hereafter, users) (n = 115). Concerns about adverse effects rose in tandem with risk factors for repeated pregnancy (r = 0.26; P = .001) and were the most important determinant of levonorgestrel implant removal (relative risk, 9.72; 95% confidence interval, 4.62-19.49). However, the number of risk factors for repeated pregnancy was also a significant, independent predictor of levonorgestrel implant removal (relative risk, 2.34; 95% confidence interval, 1.10-4.66). Following removal, contraceptive use was poor and conception occurred rapidly; 24 (37%) of the removers conceived again within 2 years of the index delivery. CONCLUSIONS: The study hypothesis was supported. Our findings that concerns about the adverse effects of levonorgestrel implants rise in tandem with risk factors for repeated pregnancy suggest that the efficacy of counseling before and after levonorgestrel implant insertion could be improved by addressing those aspects of the user's life that undermine the motivation to use contraception. PMID- 9743037 TI - The relationship of school breakfast to psychosocial and academic functioning: cross-sectional and longitudinal observations in an inner-city school sample. AB - OBJECTIVE: To determine if a relationship exists between participation in a school breakfast program and measures of psychosocial and academic functioning in school-aged children. METHODS: Information on participation in a school breakfast program, school record data, and in-depth interviews with parents and children were collected in 1 public school in Philadelphia, Pa, and 2 public schools in Baltimore, Md, prior to the implementation of a universally free (UF) breakfast program and again after the program had been in place for 4 months. One hundred thirty-three low-income students had complete data before and after the UF breakfast program on school breakfast participation and school-recorded measures, and 85 of these students had complete psychosocial interview data before and after the UF breakfast program. Teacher ratings of behavior before and after the UF breakfast program were available for 76 of these students. RESULTS: Schoolwide data showed that prior to the UF breakfast program, 240 (15%) of the 1627 students in the 3 schools were eating a school-supplied breakfast each day. Of the 133 students in the interview sample, 24 (18%) of the students ate a school supplied breakfast often, 26 (20%) ate a school-supplied breakfast sometimes, and 83 (62%) ate a school-supplied breakfast rarely or never. Prior to the UF breakfast program, students who ate a school-supplied breakfast often or sometimes had significantly higher math scores and significantly lower scores on child-, parent-, and teacher-reported symptom questionnaires than children who ate a school-supplied breakfast rarely or never. At the end of the school term 4 months after the implementation of the UF breakfast program, school-supplied breakfast participation had nearly doubled and 429 (27%) of the 1612 children in the 3 schools were participating in the school breakfast program each day. In the interview sample, almost half of the children had increased their participation. Students who increased their participation in the school breakfast program had significantly greater increases in their math grades and significantly greater decreases in the rates of school absence and tardiness than children whose participation remained the same or decreased. Child and teacher ratings of psychosocial problems also decreased to a significantly greater degree for children with increased participation in the school breakfast program. CONCLUSION: Both cross-sectional and longitudinal data from this study provide strong evidence that higher rates of participation in school breakfast programs are associated in the short-term with improved student functioning on a broad range of psychosocial and academic measures. PMID- 9743038 TI - Educational needs among pediatricians regarding caring for terminally ill children. AB - BACKGROUND: According to the Accreditation Council on Graduate Medical Education and the Ambulatory Pediatrics Association, pediatricians need to be instructed in caring for terminally ill children as a part of residency training. However, a systematic approach to education in end-of-life care is lacking in most residency programs. OBJECTIVES: To assess pediatricians' self-perception of their coping skills regarding death and dying at The New York Hospital-Cornell Medical Center, New York City, NY, if they felt further support and education were needed in this area, and which modes of instruction respondents thought would be most useful. METHODS: Data based on a questionnaire distributed to pediatricians at The New York Hospital are presented. RESULTS: The following topics are discussed: (1) existing level of education and support in caring for terminally ill patients; (2) attitudes of pediatricians regarding discussions of diagnosis and prognosis with dying children and their families; (3) pediatricians' perceived need to limit emotional involvement with terminally ill children; (4) attendance of funeral or memorial services of patients by pediatricians; (5) experience of patient death as a failure; and (6) expressed need for support and instruction in death and dying based on level of training. CONCLUSION: Educational interventions and support in this area are needed. PMID- 9743039 TI - Guides for reading and interpreting systematic reviews: III. How did the authors synthesize the data and make their conclusions? PMID- 9743040 TI - Radiological case of the month. Orange seed aspiration. PMID- 9743041 TI - Picture of the month. Half-and-half fingernails. PMID- 9743042 TI - Pathological case of the month. Pertussis pneumonia. PMID- 9743043 TI - Pharyngeal findings of group A streptococcal pharyngitis. PMID- 9743044 TI - Facial palsy in Lyme disease. PMID- 9743045 TI - Hospital infant formula discharge packages. PMID- 9743046 TI - "Bet you I will" risk. PMID- 9743047 TI - Hyponatremia in children treated with desmopressin. PMID- 9743048 TI - Neonatal circumcision. PMID- 9743049 TI - Escherichia coli and the hemolytic-uremic syndrome. AB - BACKGROUND: The hemolytic-uremic syndrome (HUS) comprises hemolytic anemia, acute renal failure, and thrombocytopenia. It is the most frequent cause of acute renal failure in children. METHODS: This review is based on an extensive overview of the literature dealing with the HUS in children. RESULTS: HUS is the most common cause of acute renal failure in infants and young children and follows a diarrheal prodrome approximately 90% of the time. Nearly all postdiarrheal cases are caused by enterohemorrhagic E coli infections, in particular serotype O157:H7. Mortality is around 5%, and approximately 50% of survivors manifest some types of sequelae. CONCLUSION: Surveillance and contact investigation are important to control outbreaks, as well as early and aggressive treatment of symptomatic subjects to prevent mortality and severe complications, such as chronic renal disease. PMID- 9743050 TI - Review of cutaneous lasers and their applications. AB - BACKGROUND: The use of lasers has assumed an increasingly important role in the treatment of a variety of cutaneous lesions over the past few decades. Because of their effectiveness, physicians from a variety of specialties have incorporated lasers into their practices. Unfortunately, widespread availability of lasers and the public's fascination with their potential uses have created extraordinary, often unrealistic, expectations. METHODS: We review the laser systems most frequently used to treat skin conditions. RESULTS: We discuss lasers with specificity for vascular malformations and pigmentary disorders as well as for tattoos and scars. Also, we review the latest techniques for cutaneous laser resurfacing with carbon dioxide and erbium:YAG lasers. Last, we briefly outline future uses of lasers and ongoing investigations, including laser treatment of leg veins and laser-assisted hair removal. CONCLUSIONS: Lasers, when properly used, offer clear advantages when compared with older, traditional approaches. Laser technology is clearly at its best when the characteristics of selectivity and specificity apply. Significant improvement and even elimination of many cutaneous lesions can now be accomplished with reduced risks to the patient when proper patient selection and laser treatment parameters are chosen. PMID- 9743051 TI - An interdisciplinary effort to help patients with limited prescription drug benefits afford their medication. AB - BACKGROUND: Patients without prescription benefits present a significant challenge to health care providers. The inability of patients to afford medication may serve as a barrier to adherence and may ultimately result in poor patient outcomes. In this report, we describe the system used at a university based adult internal medicine center to assist patients in affording medication. METHODS: Patients in need of prescription assistance are identified by any member of the interdisciplinary team, which consists of physicians, clinical pharmacists, nurses, a social worker, a financial counselor, and a dietitian. Medication profiles are reviewed to identify less expensive alternatives. State, federal, institutional, and pharmaceutical company assistance program eligibility is determined, and the appropriate program is accessed. RESULTS: The described approach is effective in assisting patients without prescription benefits with the procurement of medication. CONCLUSION: Use of this interdisciplinary approach is an innovative solution to a common problem encountered in the outpatient setting. PMID- 9743052 TI - Umbilical cord blood transplantation: providing a donor for everyone needing a bone marrow transplant? AB - BACKGROUND: Bone marrow transplantation (BMT) has been limited in the past by the availability of matched donors for patients. Over the past decade, the use of umbilical cord blood (UCB) as a source of hematopoietic stem cells has revolutionized the field of BMT, providing a source of hematopoietic stem cells for an increasing number of patients in need of a transplant. RESULTS: Umbilical cord blood transplantation (UCBT) appears to result in sustained engraftment of donor hematopoiesis similar to results achieved with marrow and peripheral blood hematopoietic stem cells. Early results indicate that UCBT is associated with a lower incidence and less severity of graft-versus-host disease than other sources of stem cells, potentially decreasing the morbidity and mortality of BMT. As the potential of UCBT has been realized, cord blood storage facilities have been established to provide UCB. The rapid emergence of UCBT has transformed a waste product of birth into a life-saving resource. Its use, however; has raised numerous ethical and medical concerns unique to this alternative source of stem cells. CONCLUSIONS: Umbilical cord blood transplantation represents a major advance in providing another stem cell source to patients in need of allogeneic hematopoietic stem cell transplantation. As with all new technologies, UCBT will have to be carefully studied over the next several years to determine its safety, efficacy, and precise indications in comparison with other sources of hematopoietic stem cells. The ethics of UCBT must properly respect the rights and needs of both donors and recipients. PMID- 9743053 TI - Hepatic amebiasis among patients in a public teaching hospital. AB - The increase in immigration to the United States is associated with diseases, such as amebiasis, that are endemic to developing countries. We retrospectively reviewed 49 public-hospital patients with hepatic amebiasis occurring between 1985 and 1995. Most patients were immigrants (47) from Latin America (43), male (43), and young (mean age, 39.8 years). Symptoms noted by more than half were abdominal pain and fever. Ultrasonography showed single lesions in 70% and right sided involvement in 85%. Serologies against Entamoeba histolytica were noted in 86%. After treatment, the median interval from admission to defervescence was 2 days, to normalization of white cell count 3 days, and to resolution of abdominal pain 4 days. Morbidity (one case of pericarditis) and mortality (one death in a cirrhotic man) were low. Hepatic amebiasis continues to be diagnosed in the United States, primarily among Hispanic and Asian immigrants. When appropriately considered, current diagnostic and therapeutic modalities result in rapid improvement and excellent outcome. PMID- 9743054 TI - Experience with clozapine in a community mental health care setting. AB - BACKGROUND: Clozapine has been heralded as the first major breakthrough in antipsychotic drug therapy for treatment-resistant schizophrenia in 40 years. This study reports on the experience with clozapine in an outpatient, community mental health care setting. METHODS: All clinic patients receiving clozapine during the 4-year period 1992 to 1996 were retrospectively studied. Measures of improvement were changes in the Clinical Global Improvement (CGI) Scale and reduction in the number of hospital days after clozapine therapy. RESULTS: Testing with the CGI scale showed moderate or marked improvement in 63% of patients. Hospital days dropped from 7,919 to 1,833 for comparable time periods. Clozapine therapy had to be discontinued in only 21% of patients, and no serious side-effects occurred. CONCLUSION: Clozapine is an effective medication for treatment-resistant schizophrenia and can be safely used in chronic mental illness. Although the drug is expensive, the cost is offset by a remarkable reduction in hospital days. PMID- 9743055 TI - Rubella serology in mentally retarded adults. AB - BACKGROUND: Rubella and congenital rubella syndrome remain a problem in the United States; institutionalized individuals are at higher risk. We assessed demographic variables and rubella serology in a population of institutionalized adults with mental retardation. METHODS: Subjects were 181 institutionalized adults with mental retardation. We reviewed charts to determine patient's age, sex, race, and fertility status (if female), cause of mental retardation, and rubella history. Serologic testing to determine rubella immunity was done. RESULTS: We found that 26.4% of subjects were nonimmune. Sex, cause of mental retardation, and history of rubella vaccination were not related to serologic immunity status. Of the 29 fertile female subjects, 8 (27.6%) were nonimmune to rubella. CONCLUSION: Subjects with unknown immunization status had a similar serologic profile to those whose medical records indicated immunization. Neither history of infection nor immunization was predictive of serology status. PMID- 9743057 TI - Admission serum albumin level and length of hospitalization in elderly patients. AB - BACKGROUND: We sought to determine whether elderly patients with a serum albumin level <3.4 g/dL on admission to community hospitals have a longer duration of hospitalization than those with an admission serum albumin level > or =3.4 g/dL. METHODS: A total of 144 patients, 60 years of age or older, were consecutively admitted to our medical service for a variety of health problems. Within 4 hours of their admission, we measured serum albumin level and recorded subsequent length of hospital stay. RESULTS: Length of hospital stay was inversely related to admission serum albumin level. The mean length of hospitalization was 3.85 +/- 2.55 days (+/- SD) for patients with an admission serum albumin level > or =3.4 g/dL and 6.74 +/- 4.79 days for those with an admission serum albumin level <3.4 g/dL. Three patients (6%) with admission serum albumin levels <3.4 g/dL died, but no deaths occurred in patients with an albumin level > or =3.4 g/dL. CONCLUSION: A serum albumin level <3.4 g/dL obtained within 4 hours of hospital admission is a reliable predictor of prolonged hospital stay and death in patients 60 years of age or older. PMID- 9743056 TI - Prostate-specific antigen values and their clinical significance in renal transplant recipients. AB - BACKGROUND: In the diagnosis of prostate cancer, prostate-specific antigen (PSA) is the most clinically useful tumor marker. Its utility in renal transplant patients is unclear. We hypothesized that PSA values might be affected by the renal function, immunosuppression, or proteinuria. METHODS: Three hundred four PSA values were measured in 166 patients >40 years (53.5 +/- 7.2 years, mean +/- SD, range 44.4 to 76.2). Charts were reviewed for 24-hour creatinine clearance, 24-hour urinary protein, cyclosporine use, age at the time of each PSA test, and evaluation done in response to the PSA result. Analyses used the Mann-Whitney U test and simple regression model. RESULTS: Twenty-five values in 13 patients were >4.0 ng/mL. Of these, 6 patients (6 values) had normal repeat PSA values; 3 patients (11 values) had negative evaluations for prostate cancer; and 4 patients (8 values) were diagnosed with prostate cancer. The latter 4 patients were excluded from the denoted normal group leaving 294 PSA values in 162 patients. The mean normal PSA was 1.3 +/- 1.8 ng/mL, range 0.1 to 20.2. The pretreatment mean PSA for recipients with prostate cancer was 302 +/- 800, range 8.0 to 2,281. An elevated PSA value was associated with a 31% incidence of prostate cancer. There was no association of PSA levels with cyclosporine use, 24-hour creatinine clearance, or 24-hour urinary protein, but there was with age. CONCLUSIONS: Prostate-specific antigen values in renal transplant recipients are similar to those in the general population and are not influenced by cyclosporine or by renal function. We recommend routine measurement of PSA in male transplant recipients. PMID- 9743058 TI - Prevalence of hot flushes during and after neoadjuvant hormonal therapy for localized prostate cancer. AB - OBJECTIVES: We sought to determine the prevalence of hot flushes after neoadjuvant hormonal therapy. METHODS: Forty-three patients who received neoadjuvant hormonal therapy before radical prostatectomy were asked to complete a questionnaire regarding hot flushes. RESULTS: Complete information was available for 35 of the 43 patients. No hot flushes were noted in 20%; in 69%, hot flushes were noted during treatment but resolved after termination of treatment; and in 11%, hot flushes continued for at least 3 months after cessation of hormonal therapy. Analyzing the data with respect to duration of hormonal therapy showed that patients receiving neoadjuvant hormonal therapy for more than 4 months had the highest incidence of persistent hot flushes. CONCLUSIONS: Hot flushes will be noted in 80% of patients who receive neoadjuvant hormonal therapy. In approximately 10%, hot flushes will continue for a significant period after hormonal therapy is terminated. Patients should be apprised of this potential side effect. PMID- 9743059 TI - Illicit alcohol (moonshine) consumption in West Alabama revisited. AB - BACKGROUND: Lacy and Wintemitz in 1984 and Pegues in 1991 showed the presence of moonshine-related lead poisoning in Alabama. METHODS: This study was a 10-year follow-up to the Lacy and Wintemitz study and used a similar inpatient retrospective chart review methodology. We looked at cases occurring between 1989 and 1992, which were positive for either a history of moonshine consumption or lead intoxication and cases suspicious for the diagnoses, based on the Lacy and Wintemitz epidemiologic parameters. RESULTS: A declining, yet continuing, presence of moonshine-related lead intoxication still exists in west Alabama. CONCLUSIONS: The findings suggest the current at-risk patients may continue to be middle-aged to elderly men from rural settings. Furthermore, appropriate screening tests were not always ordered, which suggests a possible lack of awareness of the disorder by care givers. PMID- 9743060 TI - Blastomycosis as an etiology of acute lung injury. AB - Blastomyces dermatitidis, a dimorphic broad-based budding yeast endemic to the Mississippi River Valley region, is responsible for morbidity in humans via inhalation and dissemination. The response of acute lung injury, which produces an illness with serious morbidity and an approximately 50% mortality, uncommonly occurs. Diagnosis can be difficult, and a high index of suspicion should be maintained in endemic regions for patients with acute lung injury of uncertain etiology, especially if their condition deteriorates on broad-spectrum antimicrobial and antitubercular therapy and they have a previous insidious respiratory complaint and constitutional symptoms. Diagnosis should be aggressively pursued and treatment with amphotericin B (0.6 to 0.8 mg/kg/day) initiated as early as possible. PMID- 9743061 TI - Evidence of acute stress disorder after diagnosis of cancer. AB - Cancer patients frequently have symptoms of anxiety and depression after diagnosis. Often these symptoms are apparent to the physician. We report the case of a cancer patient who appeared to her physician to be coping relatively well but was actually having psychologic symptoms that met criteria for acute stress disorder (ASD). Cancer patients who have psychologic trauma at diagnosis and meet criteria for ASD may appear to be coping better than they are. Mental health interventions for cancer patients are recommended. PMID- 9743062 TI - Symptomatic pulmonary hyalinizing granuloma. AB - An otherwise healthy 37-year-old man came to the emergency room with left-sided dull chest pain of 4 weeks' duration. Physical examination, laboratory studies, and electrocardiogram were all unremarkable. A chest x-ray film revealed calcified pulmonary nodules. Computed tomography of the chest confirmed bilateral parenchymal cavitary lesions. Via limited thoracotomy, a tan nodule measuring 2.5 to 3.0 cm in diameter was excised from the left upper lobe. Histopathologic examination revealed a well circumscribed lesion and extensive lamellar hyalinization. A few foci of finely granular calcification were present within the hyalinizing areas. After surgery and short-term use of nonnarcotic analgesics, the chest pain resolved. Although pulmonary hyalinizing granuloma (PHG) is known to produce cavitating lesions, calcification at multiple sites is also consistent with this diagnosis. Clinicians should remember to include PHG in the differential diagnosis of multiple pulmonary nodules. PMID- 9743063 TI - Leg weakness in neurosarcoidosis. AB - A 22-year-old man had leg weakness and decreased hearing. A diagnosis of neurosarcoidosis was made, based on neuroimaging studies and lymph node biopsy. Post-treatment neuroimaging, in particular magnetic resonance imaging along with clinical course, is useful in monitoring response to therapy. PMID- 9743064 TI - Simvastatin-induced lupus erythematosus. AB - We report the case of a 79-year-old man who had onset of fatigue, myalgia, and pleuritic chest pain 3 months after initiation of therapy with simvastatin. He had signs of pleuropericarditis due to simvastatin-induced lupus erythematosus. This should alert clinicians to this possible adverse effect of simvastatin and other statins. PMID- 9743065 TI - Ulnar neuropathy associated with subdermal contraceptive implant. AB - Side effects are a common occurrence in the use of subdermal contraceptive implants (Norplant); approximately 70% to 80% of women using the device report abnormal uterine bleeding, headaches, acne, mastalgia, nervousness, appetite changes, and weight gain. Local implant site reactions range from 0.4% to 4.7%, with pain being the most common. Other insertion site complications include infection and implant expulsion. Only three cases have been described in the literature concerning implant site-related neuropathy, involving the sensory branch of the musculocutaneous nerve (lateral cutaneous nerve) in two cases and the antebrachial cutaneous nerve in the third case. We believe our report is the first case of an axonal loosing motor and sensory ulnar neuropathy associated with the removal of a subdermal contraceptive implant (Norplant). We review insertion site complications and their most likely causes. Also, we discuss alternative removal techniques for difficult-to-remove implants. PMID- 9743066 TI - HIV seronegativity in an infant with the acquired immunodeficiency syndrome. AB - We report the case of an infant with progressive human immunodeficiency virus (HIV) infection and persistent seronegativity. The child had Pneumocystis carinii pneumonia at 4 months of age and was documented to be HIV-infected by HIV-1 deoxyribonucleic acid (DNA) polymerase chain reaction (PCR), but enzyme-linked immunosorbent assay (ELISA) and Western blot tests for HIV-1 and HIV-2 specific antibodies remained negative until the infant was 10 months old. This case should increase awareness about the possibility of seronegative HIV infection in infants and stress the fact that in questionable cases, even if the screening serology is negative, additional methods of diagnosis (ie, PCR, viral culture, and p24 antigen) should be considered. PMID- 9743068 TI - Creatine supplementation: safe as steak? PMID- 9743067 TI - Visceral larva migrans and the hypereosinophilia syndrome. AB - This is a case of visceral larva migrans and hypereosinophilia syndrome with persistently elevated white blood cell count despite adequate medical therapy in a 4-year-old boy with leukocytosis and splenomegaly. Medical history included reactive airway disease and geophagia (pica). Serology for Toxocara canis revealed elevated IgG and IgM titers. Ophthalmologic evaluation ruled out ocular larva migrans. After 5 days of thiabendazole therapy, leukocytosis persisted, and a second course of anthelmintics was prescribed. Two weeks later, a decrease in leukocytosis was noted. Thiabendazole therapy was continued for 15 more days. Repeated serology for T canis revealed a decreased IgM titer and a further elevated IgG titer. Follow-up showed increased physical activity, improved respiratory status, and resolution of splenomegaly. PMID- 9743069 TI - Cigar smoking. PMID- 9743070 TI - Thermoprotective mechanisms of irrigation during bipolar cautery. AB - Bipolar cautery is routinely used in operations of the head and neck, as well as in other specialties, both for dissection and for achieving hemostasis. Whereas simultaneous irrigation is frequently used to minimize neuronal injury, its effectiveness has not been tested under controlled conditions. Our objectives in this study were to test the hypothesis that including irrigation during bipolar cautery is thermoprotective and to identify the mechanisms underlying the thermoprotective effect. The thermoprotective role of irrigation with bipolar cautery was tested in a rat model in which the sciatic nerve was exposed and a 1 second stimulus at 40 or 20 watts was applied with bipolar cautery forceps placed directly on the nerve in the presence or absence of simultaneous irrigation. We used the Sciatic Functional Index as used to quantitate the degree of paresis induced. The results showed that simultaneous irrigation reduced the percentage of animals showing paresis. This effect was significant for animals exposed to 40 and 20-watt cautery. The mechanism for the reduction in the degree of paresis by irrigation could not be attributed to a lowering of the maximal temperature achieved after bipolar cautery. Instead, the thermoprotective mechanism of the irrigation involved an enhanced recovery to basal temperatures when measured at 15 seconds after nerve stimulation with 40 or 20 watts. Reducing the power from 40 watts to 20 watts did not significantly lessen the tissue temperature. The results of this study suggest that irrigation done simultaneously with bipolar cautery enhances temperature recovery to basal levels and plays a role in thermoprotection against the effects of cautery. PMID- 9743071 TI - Inhibition of nitric oxide synthase causes elevation of hearing thresholds. AB - OBJECTIVE: Nitric oxide mediates the effects of excitatory amino acids in the central nervous system. The excitatory amino acids are thought to be the neurotransmitters at the cochlear hair cell-afferent nerve synapse. Nitric oxide synthase is present in spiral ganglion cells. This study investigated the role of nitric oxide in cochlear neurotransmission. METHODS: In gerbils, cochlear compound action potential thresholds were recorded before and after cochlear perfusions with control solutions of artificial perilymph solution and test solutions of S-methyl-L-thiocitrulline (MTC), a competitive inhibitor of nitric oxide synthase. Cochleas were also preperfused with L-arginine before perfusion with a mixture of MTC/L-arginine (to overcome competitive inhibition by MTC with L-arginine, the natural substrate of nitric oxide synthase). RESULTS: Cochlear perfusion with MTC caused significant elevations of compound action potential threshold of 51 dB as opposed to insignificant elevations of only 10 dB in control animals. An insignificant threshold shift of 9 dB was observed when L arginine was coperfused with MTC. CONCLUSIONS: Nitric oxide is involved in neurotransmission/neuromodulation in the cochlea. Because nitric oxide is both a mediator of neurotoxicity and an initiator of apoptosis in the central nervous system, nitric oxide may play a role in these processes in the cochlea. PMID- 9743072 TI - Evolutionary trends in the organization of the vertebrate crista ampullaris. AB - Intraaxonal labeling studies in the toadfish, frog, turtle, and chinchilla suggest broad evolutionary trends in the vertebrate crista ampullaris. The crista of anamniotes (fish, amphibians) contains type II hair cells innervated by bouton afferents and is longitudinally organized. Type I hair cells are first seen in reptiles and birds, where they are confined to a central zone and are innervated by calyx and dimorphic afferents. The central zone is surrounded by a peripheral zone containing only type II hair cells innervated by bouton afferents. Results in the turtle suggest that the peripheral zone in reptiles and birds is organized similarly to the entire anamniote crista. The turtle central zone finds no parallel in anamniotes but resembles the mammalian central zone in its structure and afferent physiology. With the advent of a central zone in reptiles, a concentric organization is superimposed on a linearly organized peripheral zone. The mammalian crista, in contrast, has an entirely concentric organization. This may be related to the extension of the neuroepithelium further down the slopes of the crista in mammals than in other vertebrates and to the distribution of type I hair cells throughout the mammalian neuroepithelium. PMID- 9743073 TI - Hair cells in mammalian utricles. AB - Two morphological classes of mechanosensory cells have been described in the vestibular organs of mammals, birds, and reptiles: type I and type II hair cells. Type II hair cells resemble hair cells in other organs in that they receive bouton terminals from primary afferent neurons. In contrast, type I hair cells are enveloped by large cuplike afferent terminals called calyces. Type I and II cells differ in other morphological respects: cell shape, hair bundle properties, and more subtle ultrastructural features. Understanding the functional significance of these strikingly different morphological features has proved to be a challenge. Experiments that correlated the response properties of primary vestibular afferents with the morphologies of their afferent terminals suggested that the synapse between the type I hair cell and calyx ending is lower gain than that between a type II hair cell and a bouton ending. Recently, patch-clamp experiments on isolated hair cells have revealed that type I hair cells from diverse species have a large potassium conductance that is activated at the resting potential. As a consequence, the voltage responses generated by the type I hair cells in response to injected currents are smaller than those generated by type II hair cells. This may contribute to the lower gain of type I inputs to primary afferent neurons. Studies of neonatal mouse utricles show that the type I specific potassium conductance is not present at birth but emerges during the first postnatal week, a period of morphological differentiation of type I and type II hair cells. PMID- 9743074 TI - Evolution of the vestibulo-ocular system. AB - The evolutionary and developmental changes in the eye muscle innervation, the inner ear, and the vestibulo-ocular reflex are examined. Three eye muscle patterns, based on the innervation by distinct ocular motoneurons populations, can be identified: a lamprey, an elasmobranch, and a bony fish/tetrapod pattern. Four distinct patterns of variation in the vestibular system are described: a hagfish pattern, a lamprey pattern, an elasmobranch pattern, and a bony fish/tetrapod pattern. Developmental data suggest an influence of the hindbrain on ear pattern formation, thus potentially allowing a concomitant change of eye muscle innervation and ear variation. The connections between the ear and the vestibular nuclei and between the vestibular nuclei and ocular motoneurons are reviewed, and the role of neurotrophins for pattern specification is discussed. Three patterns are recognized in central projections: a hagfish pattern, a lamprey pattern, and a pattern for jawed vertebrates. Second-order connections show both similarities and differences between distantly related species such as lampreys and mammals. For example, elasmobranchs lack an internuclear system, which is at best poorly developed in lampreys. It is suggested that the vestibulo ocular system shows only a limited degree of variation because of the pronounced functional constraints imposed on it. PMID- 9743075 TI - Recording eye movements in mice: a new approach to investigate the molecular basis of cerebellar control of motor learning and motor timing. AB - The vestibulocerebellum is involved in the control of compensatory eye movements. To investigate its role in learning and timing of motor behavior, we investigated compensatory eye movements in mice with the use of search coils. Wild-type mice showed the ability to increase the gain of their vestibulo-ocular reflex by visuovestibular training. This adaptation did not occur in lurcher mice, a natural mouse mutant that completely lacks Purkinje cells. During the optokinetic reflex the phase of the eye movements of lurcher mice in reference to the stimulus lagged behind that of wild-type littermates, whereas during the vestibulo-ocular reflex it led that of the wild-type mice. During combined optokinetic and vestibular stimulation, the phase of the lurcher mice lagged behind that of the wild-type mice at the low stimulus frequencies, whereas it led the phase of the wild-type mice at the high frequencies. In addition, the optokinetic response of the lurcher mice showed a significantly longer latency during constant-velocity step stimulation than that of the wild-type mice. We conclude that Purkinje cells are necessary for both learning and timing of compensatory eye movements in mice. The present description of gain adaptation and phase dynamics provides the basis for studies in which the molecular mechanisms of cerebellar control of compensatory eye movements are investigated with the use of genetically manipulated mice. PMID- 9743076 TI - Evolution of adaptive neural networks: the role of voltage-dependent K+ channels. AB - The vestibular pathway of the mollusk Hermissenda crassicornis mediates a reflexive, unconditioned response to disorientation, clinging, that has been conserved during evolution even to the emergence of our own species. This response becomes associated with a visual stimulus (mediated by a precisely ordered visual-vestibular synaptic network) according to principles of Pavlovian conditioning that are also followed in human learning. It is not entirely surprising therefore that molecular and biophysical cascades responsible for this associative learning appear to function in both mollusks and mammals. In brief, combinational elevation of (Ca2+)i, diacylglycerol, and arachidonic acid activates protein kinase C to phosphorylate the Ca2+ and guanosine triphosphate binding protein, cp20 (now called calexcitin (Nelson T, et al. Proc Natl Acad Sci USA 1996;93:13808-13)), which potently inactivates postsynaptic voltage-dependent K+ currents and thereby increases synaptic weight. Longer term changes included rearrangement of synaptic terminals and modified protein synthesis. This cascade has also been implicated in other associative-learning paradigms (e.g., spatial maze, olfactory discrimination) and as a pathophysiologic target in early Alzheimer's disease. Recent molecular biologic experiments also demonstrate the dependence of associative memory (but not long-term potentiation) on voltage dependent K+ currents. Theoretic learning models based on these findings focus on dendritic spine clusters and yield computer implementations with powerful pattern recognition capabilities. PMID- 9743077 TI - Role of the flocculus of the cerebellum in motor learning of the vestibulo-ocular reflex. AB - Structure-function studies at the systems level are an effective method for understanding the relationship of the central nervous system to behavior. Motor learning or adaptation of the vestibulo-ocular reflex is a clear example wherein this approach has been productive. During a vestibulo-ocular reflex the brain converts a head velocity signal, transduced through the vestibular semicircular canals, into an eye movement command delivered to the extraocular muscles. If the viewed target remains on the fovea of the retina, the reflex is compensatory, and its gain, eye velocity/head velocity, is one. When the image of the viewed object slips across the retina, visual acuity decreases, and the gain of the reflex, which is no longer one, is plastically adapted or adjusted until retinal stability is restored. The anatomic substrate for this plasticity thus involves brain structures in which visual-vestibular interaction can potentially occur, as well as vestibular and visual sensory and oculomotor motor structures. Further, it has been known for many years that removal of the flocculus of the cerebellum permanently precludes further vestibulo-ocular reflex adaptation, demonstrating the involvement of the cerebellum in this behavior. Maekawa and Simpson (J Neurophysiol 1973;36: 649-66) discovered that one visual input to the flocculus involved the accessory optic system and the inferior olive. Ensuing work has demonstrated that the visual signals used to adapt the vestibulo-ocular reflex are transmitted by this accessory optic system to the flocculus and subsequently to brain stem structures involved in vestibulo-ocular reflex plasticity. Presently the inclusive list of anatomic sites involved in vestibulo-ocular reflex circuitry and its adaptive plasticity is small. Our laboratory continues to believe that this behavior should be caused by interactions within this small class of neurons. By studying each class of identified neuron and its interactions with others within the list, we hope to ultimately understand the mechanisms used by the brain in the expression of this behavior. PMID- 9743078 TI - Output-to-input approach to neural plasticity in vestibular pathways. AB - Some thoughts on current interpretations of available data regarding vestibular compensation at functional, network, and neural levels are presented. Basic concepts related to neural plasticity (or elasticity) underlying motor learning and regeneration also are discussed briefly. Modifiability in vestibular pathways, at both the functional and structural levels, after peripheral and central axotomy, and subsequent to transient or permanent chemical target removal, is presented as an experimental ground to explain similarities and differences between regenerative, compensatory, and adaptive mechanisms in the mammal central nervous system. PMID- 9743079 TI - Vestibular adaptation: how models can affect data interpretations. AB - Vestibular adaptation can be induced optically or by chemical or physical injury to the vestibular apparatus or the brain stem. In searching for the sites or mechanisms of vestibular adaptation, neurophysiologists often rely on comparing central resting (background) activities and central modulations (sensitivity) during vestibular stimulation, before and after motor learning or vestibular compensation. It is assumed that adapted central sites must exhibit modulation changes that parallel vestibulo-ocular reflex changes. Using model simulations and analysis, we will show that such presumptions may be misleading. First, using a simple schematic of interconnected cells or nuclei, one can show that modulation depth and background "tone" can be modified (or fixed) independently, using weightings on direct or indirect afferent projections. That is, if synaptic weights along all stimulus pathways are altered, one may fix or strongly modify central premotor characteristics in a manner apparently unrelated to global reflex changes. In the vestibulo-ocular reflex, the dominant premotor pathways contain position-vestibular-pause cells and eye-head-velocity cells (which are behaviorally similar to floccular-target neurons). Several experiments have reported negligible changes in the velocity sensitivity of position-vestibular pause cells, despite large gain changes in the vestibulo-ocular reflex induced by training with visual-vestibular conflict. On the other hand, the modulation changes on floccular-target neurons (position-vestibular-pause) can be much larger than the changes in reflex gain. Using a bilateral vestibulo-ocular reflex model, we show that overall increases or decreases in reflex gain can be expressed (even overexpressed) in one particular subgroup of premotor neurons. Nevertheless, such observations are theoretically compatible with synaptic changes on all primary projections in a widely interconnected central network. Hence, stable neural responses during reflex adaptation are not sufficient to exclude a potential site of sensory-motor adaptation. Similarly, modified neural responses (as in cerebellum) need not necessarily imply a direct role in supporting the adapted state. Model predictions should help to design additional experimental protocols, to test hypotheses, and to refine diagnostic measures of recovery after vestibular lesions. PMID- 9743080 TI - Vestibular nerve regeneration in the bullfrog, Rana catesbeiana: peripheral dendrites. AB - Three experiments were conducted on healthy adult bullfrogs (Rana catesbeiana) for the purpose of investigating three characteristics of centrifugal vestibular afferent regeneration after complete transection of the anterior division of the vestibular nerve (AVN). In experiment 1 total fiber count and axon diameter measurements were obtained from the anterior canal nerve at three different time periods and compared with normal. The normal group (n = 3) demonstrated a total fiber count of 1001 +/- 76 (SEM). The early time period (1 to 2 weeks, n = 3) did not completely regenerate as demonstrated by a total fiber count of 282 +/- 23. The intermediate (4 to 6 weeks, n = 3) and late (8 to 16 weeks, n = 3) groups exhibited total fiber counts of 907 +/- 29 and 946 +/- 50, respectively, which were not different from normal (Mann-Whitney U, p > 0.2). Evaluation of fiber diameter distribution of the intermediate and late regenerated nerves revealed a reduction in axon diameter caliber compared with normal (analysis of variance, p < 0.0001). Thus transection of the AVN results in regeneration of all afferents that exhibit a reduction in axon diameter. In experiment 2 fibers innervating the anterior canal crista (ACC) were prelabeled before nerve transection. After the labeling procedure the AVN (n = 3) was sectioned at a location that resulted in denervation of three vestibular receptors: the ACC, horizontal canal cristae (HCC), and utricular macula. After 4 weeks of regeneration the ACC fibers that were prelabeled were observed innervating all three denervated vestibular receptors. This result demonstrated that reinnervation of the peripheral vestibular end organs after AVN transection is a nonspecific process. In experiment 3, 167 regenerated canal afferents were evaluated for functional recovery 16 weeks after transection. Both spontaneous and rotation-induced discharge characteristics were obtained and compared with those obtained from a sample of 254 normal afferents in a previous study (Hoffman LF. Factors affecting the response dynamics of canalicular primary afferent neurons in the bullfrog. St. Petersburg (FL): Association for Research in Otolaryngology; 1989). The mean spontaneous discharge coefficient of variation (CV) +/- standard deviation was 0.60 +/- 0.32 and 0.49 +/- 0.33 for ACC and HCC regenerated afferents, respectively, which did not differ from the normal means of 0.63 +/- 0.33 and 0.54 +/- 0.36 (Mann-Whitney, p > 0.2). Response gains and phases obtained during 0.05 Hz sinusoid rotations at 15 degrees/second maximum horizontal table velocity also demonstrated normal discharge characteristics. The mean phases were -28.2 +/ 25.2 degrees and -55.9 +/- 21.5 degrees for regenerated ACC and HCC afferents, respectively, which were not different from the normal means of -33.77 +/- 24.31 degrees and -58.0 +/- 23.3 degrees (Mann-Whitney U). Furthermore, regenerated afferents exhibited a positive association between phase and CV, which was also true for normal afferents (correlation analysis, p > 0.001). Although the mean gains for regenerated ACC and HCC (7.13 +/- 5.5 and 3.3 +/- 2.4 spikes x sec( 1)/degrees x sec(-2), respectively) afferents were reduced from normal ACC and HCC (14.8 +/- 12.52 and 7.76 +/- 6.58 spikes x sec(-1)/degrees x sec(-2), respectively) afferents (Mann-Whitney U, p > 0.0001), a positive association between gain and CV was also demonstrated by regenerated afferents, as was the case for normal afferents (correlation analysis, p < 0.001). Thus the overall response discharges of regenerated afferents were comparable with normal afferents. Normally, large fibers innervate central regions of the receptor, and smaller fibers innervate the peripheral regions. However, the data from experiments 1 and 2 demonstrate that vestibular nerve regeneration results in a dissociation between the normal topographic organization of fiber size and regional innervation of the receptor epithelium. (ABSTRACT TRUNCATED) PMID- 9743081 TI - Hair cell recovery in the chinchilla crista ampullaris after gentamicin treatment: a quantitative approach. AB - The mechanisms for hair cell recovery were investigated after intraortic application of 50 microg gentamicin into the perilymphatic space of the superior semicircular canal of the chinchilla. Histologic evaluation of one normal group and four posttreatment groups (7, 14, 28, and 56 days) was made with light and transmission electron microscopic techniques. The numeric changes of hair cells and supporting cells was quantified with the dissector technique. At 7 and 14 days after treatment, no type I hair cells were present, and 85% and 88% of type II hair cells were lost. Supporting cells decreased to 76% at 7 days, but they recovered to 91% at 14 days. Recovery of the epithelia was evident 28 days after treatment; 83% were type II hair cells, and 3% were type I hair cells. The supporting cell number remained close to normal (86%). Between 14 and 28 days after treatment, there was an increase of 1758 of type II hair cells, representing approximately 125 new hair cells per day. At the same time interval the number of supporting cells remained near normal. These results suggest that new hair cells might be the result of supporting cell mitotic division and differentiation. PMID- 9743082 TI - From genes to behavior in the vestibular system. AB - The central nervous system of all vertebrate embryos is derived from a series of conspicuous segments, called neuromeres, that are particularly visible in the midbrain and hindbrain areas, giving rise to the brain stem sensory and motor nuclei. This article focuses on a series of eight embryonic rhombomeric segments whose progeny can be identified in adults by the locations of iteratively homologous reticulospinal neurons and cranial motor nuclei IV through XII. Evidence shows that these rhombomeric units represent domains of gene expression, lineage restriction, and accordingly, individual vestibular neuronal phenotypes with unique oculomotor and spinal projections. Preliminary electrophysiologic and behavioral correlates of a few vestibulo-oculomotor subgroups are used as examples to illustrate the hypothesis that homologous vestibular phenotypes likely exist in all taxa because the genetic prepattern is already well established in primitive vertebrates. Finally, the segmented hindbrain arrangement responsible for the longitudinally arranged column of vestibular subnuclei is placed in perspective with genetic and molecular approaches that will eventually permit a causal reconstruction of the signaling mechanisms responsible for the development of unique vestibular subgroups. PMID- 9743083 TI - Laryngeal hematoma after thrombolytic therapy for myocardial infarction. PMID- 9743084 TI - Nitric oxide is a regulator of mucociliary activity in the upper respiratory tract. AB - The in vitro effects of the nitric oxide (NO) substrate L-arginine on ciliary beat frequency and the in vivo effects of the NO donor sodium nitroprusside (SNP) on mucociliary activity were investigated in the rabbit maxillary sinus mucosa with photoelectric techniques. L-Arginine increased ciliary beat frequency in vitro with a maximum response of 27.1% +/- 6.4% at 10(-3) mol/L, and this effect was reversibly blocked by pretreatment with the NO synthase (NOS) inhibitor N(G) nitro-L-arginine, whereas D-arginine had no such effect. SNP increased mucociliary activity in vivo, the peak response of 36.8% +/- 4.2% being obtained at the dose of 30.0 microg/kg. No tachyphylaxis was observed after repeat challenge with SNP. The increase in mucociliary activity caused by SNP was largely unaffected by pretreatment with the calcium channel blocker nifedipine, the cyclooxygenase inhibitor diclofenac, and the cholinergic antagonist atropine. The nonselective beta-blocker propranolol delayed the peak response of SNP to 7 to 8 minutes after challenge, compared with 1 to 2 minutes after challenge in animals without pretreatment. The results show the NO substrate L-arginine and the NO donor SNP to have ciliostimulatory effects in vitro and in vivo, respectively. The occurrence of NOS production in the sphenopalatine ganglion and sinus mucosa of the rabbit was studied by immunohistochemistry for NOS activity or nicotinamide adenine dinucleotide phosphate-diaphorase histochemistry. The latter is an indirect sign of neuronal NOS activity. Numerous NOS-containing cell bodies were seen in the sphenopalatine ganglion; in the sinus mucosa a moderate supply of thin NOS-immunoreactive nerve fibers was seen. Taken together, the morphologic findings and the functional results indicate NO to be a regulator of mucociliary activity in upper airways. PMID- 9743085 TI - Palatoplasty for snoring: a randomized controlled trial of three surgical methods. AB - In a randomized, controlled trial, 62 patients (47 men and 15 women) with severe antisocial snoring, but no sleep apnea, were allocated to one of three surgical treatments. These were uvulopalatopharyngoplasty, laser palatoplasty, and diathermy palatoplasty. Postoperative morbidity was measured on a visual analogue scale of severity of pain, dysphagia, and nasal regurgitation at 1, 2, and 7 days after the operation. Efficacy of each procedure was measured by asking the sleeping partner to record the severity of snoring before and after the operation, again on a visual analogue scale. Measurements were taken at 1, 3, and 6 months. There were no significant differences in early postoperative morbidity among the treatment groups. Diathermy palatoplasty is a new technique for the relief of snoring that is associated with low morbidity and requires little in the way of expensive equipment. PMID- 9743086 TI - Hereditary hemorrhagic telangiectasia. PMID- 9743087 TI - Atypical tuberculosis (nontuberculous mycobacterium) of the parotid region in children. PMID- 9743088 TI - Testicular cancer. AB - The following article provides a comprehensive review of male germ cell tumors; the pathology and the clinical manifestations of the tumors are discussed, as are the modern concepts of clinical staging. Patients with bulky stage II and stage III non-seminomatous germ cell tumors are treated with chemotherapy. The new international classification system has provided a very useful way to categorize these patients by prognosis. Patients with good- or intermediate-risk tumors may be treated with 3 courses of cisplatin, etoposide, and bleomycin (BEP) or 4 courses of etoposide and cisplatin (EP), and more than 90% of these patients will survive. Randomized trials have shown that, if only 3 courses of chemotherapy are to be given, the substitution of carboplatin for cisplatin and the omission of bleomycin are deleterious to outcome. Patients who still have a significant residual mass and normal markers after treatment should undergo a surgical resection of the residual tumor. Patients who are classified by the international classification system as having poor-risk tumors have about a 50% likelihood of survival, and many of these patients will require surgical resection of a residual tumor after chemotherapy. No randomized trial has proved a regimen to be superior to that of 4 courses of BEP. Currently, an ongoing trial is evaluating the effect of the early use of high-dose therapy in combination with hematopoietic rescue in patients with these types of tumors. Patients with small volume stage II tumors are generally treated with retroperitoneal lymph node dissection (RPLND). About 25% of the patients selected for this procedure will actually have pathologically negative nodes. Those with positive nodes may elect to receive adjuvant chemotherapy (2 courses of BEP), which will almost always prevent relapse. An alternate approach for patients willing to comply with monthly follow-up is surveillance, with chemotherapy deferred until relapse is noted. About 50% of these patients will be cured with surgery (as many as 75% have microscopic disease only). With careful follow-up, those destined to relapse can be treated promptly and at a time when they have small-volume tumors and an excellent prognosis if they go on to receive chemotherapy. Patients with clinical stage I nonseminomatous germ cell tumors may also undergo RPLND, although an acceptable alternative for these patients is surveillance. The advantages and the disadvantages of each approach are discussed. The overall risk of recurrence is about 30%, but there have been patient groups defined that may vary in risk from 10% to 15% up to 50% or more. Patients with advanced seminoma are treated with chemotherapy. When this procedure is used, outcomes are favorable and all patients are either in good- or intermediate-risk groups, according to the international classification system. Patients with small-volume stage II tumors are treated with radiotherapy. Radiation is also generally used for the treatment of clinical stage I patients, although surveillance is growing in prominence as a means to treat these patients. Late effects of treatment are also discussed in this article. Ejaculatory function can be preserved in most patients who have early stage tumors and who undergo RPLND and in some patients who undergo surgery after chemotherapy. The most troubling effect of chemotherapy is the development of etoposide-induced leukemia, a unique--and fortunately rare--clinical entity. PMID- 9743090 TI - Beat bioterror with batch science. PMID- 9743091 TI - CYTED-BT: an international biotechnology network that works. Program of Science and Technology for Development. PMID- 9743089 TI - A second Human Genome Project? PMID- 9743093 TI - Big business is high on the Honolulu method. PMID- 9743092 TI - ONSA, the Sao Paulo Virtual Genomics Institute. Organization for Nucleotide Sequencing and Analysis. PMID- 9743094 TI - On human cloning. PMID- 9743095 TI - No prokaryotic GPI anchoring. PMID- 9743097 TI - Pharming seeks public support. PMID- 9743096 TI - Centaur abandons US for European IPO. Initial Public Offerings. PMID- 9743098 TI - Cloning improvements suggested. PMID- 9743099 TI - Competition drives agriculture's genomics deals. PMID- 9743100 TI - Mystery plea for review of seed and biotech mergers. PMID- 9743101 TI - Generous German funding helps Ribozyme. PMID- 9743102 TI - United Nations to help Cuba sell biotechnology. PMID- 9743103 TI - Europe proposes orphan drug legislation. PMID- 9743104 TI - Roche integrates HIV therapeutics and diagnostics. PMID- 9743105 TI - Xenophobia quelled in the UK. PMID- 9743106 TI - New "deCODE bill" restarts controversy. PMID- 9743107 TI - Agrobacterium T-DNA: a silver bullet for filamentous fungi? PMID- 9743108 TI - Naked DNA dons new clothes. PMID- 9743109 TI - Peptide nucleic acid smugglers. PMID- 9743110 TI - Genomic microbiology: right on target? PMID- 9743111 TI - A convert to fructans in sugar beet. PMID- 9743112 TI - Array of hope for biosensors in Europe. PMID- 9743113 TI - Laboratory firepower for infectious disease research. PMID- 9743114 TI - From bits to bases: computing with DNA. PMID- 9743115 TI - Stress-inducible responses and heat shock proteins: new pharmacologic targets for cytoprotection. AB - Molecular chaperones protect proteins against environmental and physiologic stress and from the deleterious consequences of an imbalance in protein homeostasis. Many of these stresses, if prolonged, result in defective development and pathologies associated with a diverse array of diseases due to tissue injury and repair including stroke, myocardial reperfusion damage, ischemia, cancer, amyloidosis, and other neurodegenerative diseases. We discuss the molecular nature of the stress signals, the mechanisms that underlie activation of the heat shock response, the role of heat shock proteins as cytoprotective molecules, and strategies for pharmacologically active molecules as regulators of the heat shock response. PMID- 9743116 TI - Agrobacterium tumefaciens-mediated transformation of filamentous fungi. AB - Agrobacterium tumefaciens transfers part of its Ti plasmid, the T-DNA, to plant cells during tumorigenesis. It is routinely used for the genetic modification of a wide range of plant species. We report that A. tumefaciens can also transfer its T-DNA efficiently to the filamentous fungus Aspergillus awamori, demonstrating DNA transfer between a prokaryote and a filamentous fungus. We transformed both protoplasts and conidia with frequencies that were improved up to 600-fold as compared with conventional techniques for transformation of A. awamori protoplasts. The majority of the A. awamori transformants contained a single T-DNA copy randomly integrated at a chromosomal locus. The T-DNA integrated into the A. awamori genome in a manner similar to that described for plants. We also transformed a variety of other filamentous fungi, including Aspergillus niger, Fusarium venenatum, Trichoderma reesei, Colletotrichum gloeosporioides, Neurospora crassa, and the mushroom Agaricus bisporus, demonstrating that transformation using A. tumefaciens is generally applicable to filamentous fungi. PMID- 9743117 TI - High level fructan accumulation in a transgenic sugar beet. AB - We have transformed sugar beet into a crop that produces fructans. The gene encoding 1-sucrose:sucrose fructosyl transferase (1-SST), which was isolated from Helianthus tuberosus, was introduced into sugar beet. In H. tuberosus, 1-SST mediates the first steps in fructan synthesis through the conversion of sucrose (GF) into low molecular weight fructans GF2, GF3, and GF4. In the taproot of sugar beet transformed with the 1-sst gene, the stored sucrose is almost totally converted into low molecular weight fructans. In contrast, 1-sst expression in the leaves resulted in only low levels of fructans. Despite the storage carbohydrate having been altered, the expression of the 1-sst gene did not have any visible effect on phenotype and did not affect the growth rate of the taproot as observed under greenhouse conditions. PMID- 9743118 TI - Large-scale production of UDP-galactose and globotriose by coupling metabolically engineered bacteria. AB - A large-scale production system of uridine 5'-diphospho-galactose (UDP-Gal) has been established by the combination of recombinant Escherichia coli and Corynebacterium ammoniagenes. Recombinant E. coli that overexpress the UDP-Gal biosynthetic genes galT, galK, and galU were generated. C. ammoniagenes contribute the production of uridine triphosphate (UTP), a substrate for UDP-Gal biosynthesis, from orotic acid, an inexpensive precursor of UTP. UDP-Gal accumulated to 72 mM (44 g/L) after a 21 h reaction starting with orotic acid and galactose. When E. coli cells that expressed the alpha1,4-galactosyltransferase gene of Neisseria gonorrhoeae were coupled with this UDP-Gal production system, 372 mM (188 g/L) globotriose (Galalpha1-4Galbeta1-4Glc), a trisaccharide portion of verotoxin receptor, was produced after a 36 h reaction starting with orotic acid, galactose, and lactose. No oligosaccharide by-products were observed in the reaction mixture. The production of globotriose was several times higher than that of UDP-Gal. The strategy of producing sugar nucleotides by combining metabolically engineered recombinant E. coli with a nucleoside 5'-triphosphate producing microorganism, and the concept of producing oligosaccharides by coupling sugar nucleotide production systems with glycosyltransferases, can be applied to the manufacture of other sugar nucleotides and oligosaccharides. PMID- 9743119 TI - A genome-based approach for the identification of essential bacterial genes. AB - We have used comparative genomics to identify 26 Escherichia coli open reading frames that are both of unknown function (hypothetical open reading frames or y genes) and conserved in the compact genome of Mycoplasma genitalium. Not surprisingly, these genes are broadly conserved in the bacterial world. We used a markerless knockout strategy to screen for essential E. coli genes. To verify this phenotype, we constructed conditional mutants in genes for which no null mutants could be obtained. In total we identified six genes that are essential for E. coli (yhbZ, ygjD, ycfB, yfil, yihA, and yjeQ). The respective orthologs of the genes yhbZ, ygjD, ycfB, yjeQ, and yihA are also essential in Bacillus subtilis. This low number of essential genes was unexpected and might be due to a characteristic of the versatile genomes of E. coli and B. subtilis that is comparable to the phenomenon of nonorthologous gene displacement. The gene ygjD, encoding a sialoglycoprotease, was eliminated from a minimal genome computationally derived from a comparison of the Haemophilus influenzae and M. genitalium genomes. We show that ygjD and its ortholog ydiE are essential in E. coli and B. subtilis, respectively. Thus, we include this gene in a minimal genome. This study systematically integrates comparative genomics and targeted gene disruptions to identify broadly conserved bacterial genes of unknown function required for survival on complex media. PMID- 9743120 TI - Cell penetrating PNA constructs regulate galanin receptor levels and modify pain transmission in vivo. AB - Peptide nucleic acids (PNAs) form stable and tight complexes with complementary DNA and/or RNA and would be promising antisense reagents if their cellular delivery could be improved. We show that a 21-mer PNA, complementary to the human galanin receptor type 1 mRNA, coupled to the cellular transporter peptides, transportan or pAntennapedia(43-58), is efficiently taken up into Bowes cells where they block the expression of galanin receptors. In rat, the intrathecal administration of the peptide-PNA construct results in a decrease in galanin binding in the dorsal horn. The decrease in binding results in the inability of galanin to inhibit the C fibers stimulation-induced facilitation of the rat flexor reflex, demonstrating that peptide-PNA constructs act in vivo to suppress expression of functional galanin receptors. PMID- 9743121 TI - Functional gene transfer from intracellular bacteria to mammalian cells. AB - We provide evidence of direct transfer of functional DNA from bacteria to mammalian cells. An Escherichia coli K12 diaminopimelate auxotroph made invasive by cloning the invasin gene from Yersinia pseudotuberculosis transfers DNA after simple co-incubation, into a variety of mammalian cell lines. Transfer efficiency was enhanced in some cells by coexpression of the gene for listeriolysin from Listeria monocytogenes. Expression of the acquired genes occurs in both dividing and quiescent cells. The only requirement for bacteria to transfer genetic material into nonprofessional phagocytic cells and macrophages is the ability to invade the host cell. PMID- 9743122 TI - Gene transfer into muscle by electroporation in vivo. AB - Among the nonviral techniques for gene transfer in vivo, the direct injection of plasmid DNA into muscle is simple, inexpensive, and safe. Applications of this method have been limited by the relatively low expression levels of the transferred gene. We investigated the applicability of in vivo electroporation for gene transfer into muscle, using plasmid DNA expressing interleukin-5 (IL-5) as the vector. The tibialis anterior muscles of mice were injected with the plasmid DNA, and then a pair of electrode needles were inserted into the DNA injection site to deliver electric pulses. Five days later, the serum IL-5 levels were assayed. Mice that did not receive electroporation had serum levels of 0.2 ng/ml. Electroporation enhanced the levels to over 20 ng/ml. Histochemical analysis of muscles injected with a lacZ expression plasmid showed that in vivo electroporation increased both the number of muscle fibers taking up plasmid DNA and the copy number of plasmids introduced into the cells. These results demonstrate that gene transfer into muscle by electroporation in vivo is more efficient than simple intramuscular DNA injection. PMID- 9743123 TI - A rational design strategy for protein hormone superagonists. AB - By combining evolutionary considerations, sequence comparisons and homology modeling we have designed recombinant human thyroid-stimulating hormone (hTSH) analogs with increased receptor binding and activity. The introduction of seven basic residues into the peripheral loops of hTSH resulted in up to a 50,000-fold increase in receptor binding affinity and 1300-fold increase in intrinsic activity. Such analogs are not only of potential clinical interest but can be tools to explore molecular aspects of conventional as well as nonclassical actions of glycoprotein hormones. These design strategies should be applicable to the development of novel analogs of other related hormones and growth factors with a variety of therapeutic and basic science applications, particularly for proteins that have undergone evolutionary decrease in bioactivity. PMID- 9743124 TI - Patenting DNA sequences. PMID- 9743125 TI - Resources for molecular biology. PMID- 9743127 TI - Have we forgotten our basics? PMID- 9743126 TI - Complement inhibitors. PMID- 9743128 TI - Effect of an acidic primer on shear bond strength of orthodontic brackets. AB - A unique characteristic of some new etching systems is that they combine the conditioning and priming agents into a single acidic primer solution. The purpose of this study was to determine the effects on the shear bond strength and the bracket/adhesive failure mode when an acidic primer and other enamel etchants were used to condition the enamel surface before bonding. The brackets were bonded to extracted human teeth according to one of four protocols following the manufacturers instructions. Group I, teeth were etched with 37% phosphoric acid, the brackets were then bonded with System 1+ adhesive (Ormco Corporation. Orange, Calif.); group II, teeth were etched with 10% maleic acid, the brackets were also bonded with System 1+ adhesive; group III, an acidic primer that contains both the acid (phenyl-P) and the primer (hema and dimethacrylate) were placed on the enamel for 30 seconds. The adhesive used on this group was a lightly filled resin that contains Bis-GMA and HEMA. (Clearfil Liner Bond 2. J.C. Moritta, Kuraway, Japan); Group IV, the same acidic primer was used as in group II, the adhesive used was highly filled (Panavia 21. J.C. Moritta) and contains Bis-GMA. The present in vitro findings indicated that the use of acidic primers to bond orthodontic brackets to the enamel surface could provide clinically acceptable shear bond forces (x = 10.4 +/- 4.4 MPa) when used with a highly (77%) filled adhesive (Panavia 21). These debonding forces were comparable to those obtained when the enamel was conditioned with either Phosphoric (x = 11.8 +/- 4.1 MPa) or Maleic (x = 10.9 +/- 4.4 MPa) acids. With the use of a lightly (10%) filled adhesive (Clearfil Liner Bond 2), the shear bond strength was significantly lower (x = 5.9 +/- 5.6 MPa). It is of interest to note that there was a tendency to have less residual adhesive remaining on the tooth when an acid primer was used than when phosphoric and maleic acids were used. This might be of advantage to the clinician because it will require less time to clean the teeth after debonding. PMID- 9743130 TI - Ideal arch force systems: a center-of-resistance perspective. AB - The analysis of force systems from an ideal arch has shown that the ratio of the moments produced by a straight wire connecting two malaligned brackets depends on the ratio of the angulations of the brackets to the interbracket axis. Although this result permits assessment of the relative forces and moments, prediction of future tooth movement requires knowledge of the position of center of resistance as well. In this study, the forces and moments produced by a straight portion of an arch wire were transferred from the brackets to the center of resistance. The purpose was to compare the force system at the brackets to the force system at the center of resistance and to assess whether bracket geometry can be applied to predict initial tooth movement. A computer model was used to simulate two teeth connected by a straight portion of wire. Forces and moments were calculated with the use of equations derived from elementary beam theory. The results show that the force system at the center of resistance may be of an entirely different "geometry" type than that at the bracket. Factors that influence the force system include the interbracket distance, the angulation of the teeth, the length of the tooth root, and the width of the bracket. PMID- 9743129 TI - Lateral cephalometric analysis of asymptomatic volunteers and symptomatic patients with and without bilateral temporomandibular joint disk displacement. AB - Few studies of dentofacial and orthodontic structural relationships relative to temporomandibular joint (TMJ) dysfunction have been reported. We undertook this investigation to determine any correlation of orthodontic and dentofacial characteristics with TMJ bilateral disc displacement. The population of patients was selected from a TMJ clinic where a control group of asymptomatic volunteers had been previously established and standardized. Differences in skeletal structural features were determined among three study groups: (1) asymptomatic volunteers with no TMJ disk displacement, (2) symptomatic patients with no TMJ disc displacement, and (3) symptomatic patients with bilateral TMJ disk displacement. Thirty-two asymptomatic volunteers without disk displacement (25 female, 7 male) were compared with the same number each of symptomatic patients without TMJ disk displacement and symptomatic patients with bilateral TMJ disk displacement. All subjects had undergone a standardized clinical examination, bilateral TMJ magnetic resonance imaging, and lateral cephalometric radiographic analysis. The groups were matched according to sex, TMJ status, age, and Angle classification of malocclusion. Seventeen lateral cephalometric radiographic cranial base, maxillomandibular, and vertical dimension variables were evaluated and compared among the study groups. The mean angle of SNB, or the intersection of the sella-nasion plane and the nasion-point B line (indicating mandibular retrognathism relative to cranial base), of the symptomatic patients-with displacement group was significantly smaller than that in the asymptomatic volunteers and symptomatic patients without bilateral disk displacement (p < 0.05). Female subjects showed smaller linear measurements of mandibular length, lower facial height, and total anterior facial height than male subjects in all three groups (p < 0.05). The mean angle of ANB, or the intersection of the nasion point A and nasion-point B planes (indicating retrognathism of mandible relative to maxilla), was significantly greater in female than in male subjects, in all groups (p < 0.05). Symptomatic patients with bilateral disk displacement had a retropositioned mandible, indicated by a smaller mean SNB angle compared with that in asymptomatic volunteers and symptomatic patients with no disk displacement on either side. Lateral cephalometric radiographic assessment may improve predictability of TMJ disk displacement in orthodontic patients but is not diagnostic; nor does the assessment explain any cause-and-effect relationship. PMID- 9743131 TI - A comparative study of facial profiles in extraction and nonextraction treatment. AB - Evaluation of facial profiles and facial balance is a constant, continuous study and learning process for orthodontists. Tooth movement and proper positioning of the teeth to ensure favorable facial changes and to avoid unfavorable changes should be in the orthodontist's "diagnostic" mind from the very first examination. The continuous controversy concerning extraction of premolar teeth and its supposed effects on the facial profile led to the development of this study, which compares the pretreatment and posttreatment facial profiles of patients who underwent premolar extraction with those of patients who did not undergo extraction. The study comprised 170 consecutively treated patients. One hundred-eight of the patients were treated with premolar extraction, and 62 were treated without premolar extraction. The diagnostic extraction and treatment decisions were based on a total space analysis with differential extraction protocols. The Z-angle (an angular measurement) and the E-value (a linear measurement) were used to quantify and compare the pretreatment and posttreatment profiles of the two groups. Both groups completed treatment within the normal ranges of the profile measurement values with the lip profile position slightly more retrusive in the nonextraction group. The nonextraction group's posttreatment Z-angle was 5.27 degrees greater and the posttreatment E-value was 1.47 mm more negative than that of the extraction group. The extraction group began treatment with greater facial imbalance and had the greatest improvement in facial esthetics. PMID- 9743132 TI - Force distribution of the temporomandibular joint and temporal bone surface subjected to the head-chincup force. AB - The present study is an in vitro study of chincup therapy. The purpose of the study was to investigate the localization and distribution of stress induced by the head-chincup appliance. Thin single and three-dimensional strain gauges were affixed on a young dry human skull capped by the head-chincup appliance. Three kilograms of force was then applied in the direction of the condyle. In a slight opening (5 mm at the incisor position), compressive force was observed to be constant at the anterior part of mandibular neck and tensile force at the posterior part. The joint cavity showed both tensile and compressive forces, whereas its posterior site showed only compressive force. Stress distribution at the lateral surface of the temporal bone indicated that long-term use of the chincup appliance affects the craniofacial structures. The longitudinal laminagraph records of a clinical case were also presented to support the current biomechanical findings. PMID- 9743133 TI - Multivariate prediction of skeletal Class II growth. AB - Prediction of craniofacial growth is one of the keys to successful orthodontic treatment and stability. Despite numerous attempts at growth forecasting, our ability to accurately predict growth is limited. The present study outlines a possible new approach to prediction of craniofacial growth that differs from any previous attempt because of both the methods used and type of patients studied. The purpose of this study is to create and test prediction equations for forecasting favorable or unfavorable patterns of growth in skeletal Class II preadolescents. The subjects for this study include 19 females and 12 males from the Bolton growth center in Cleveland, Ohio. The patients were all untreated orthodontically, had lateral cephalometric headfilms taken biannually from the ages of 6 through 18 and had a Class II skeletal relationship at age 8. Twenty six skeletal and dental landmarks were identified and digitized, and 48 linear, angular, and proportional measurements were calculated. The subjects were divided into two groups based on anterior-posterior changes in the relationship between the maxilla and mandible. Eleven patients were in the favorable growth group, with an average improvement of 4.13 degrees in the ANB angle; 20 patients were in the unfavorable growth group with an average increase of 0.16 degrees in the ANB angle. The following prediction formula was created with Bayes theorem and assuming a multivariate Gaussian distribution: P(Good?Fn) = ke (-(0.5)) ?Fn - mu(ng)?sigma(g)(-1)?Fn - mu(ng)?T. The equation's sensitivity and specificity was calculated from serial cephalometric data from ages 6, 8, 10, and 12. The results obtained with this equation indicate 82.2% sensitivity, 95% specificity with a overall positive predictive value of 91%. This corresponds to 17.8% of patients being incorrectly identified as Poor Growers and only 5% of our patients were incorrectly identified as Good Growers. We conclude that this prediction formula improves the ability to predict favorable or unfavorable patterns of growth in this sample of skeletal Class II preadolescents. PMID- 9743134 TI - Reactions of peri-implant tissues to continuous loading of osseointegrated implants. AB - Functional and morphologic reactions of peri-implant bone surrounding screw implants (Bonefit) were studied in three dogs by loading the implants with continuous forces of 2 (about 204 gm) and 5 N (about 510 gm). Eight implants were inserted to an endosseous length of 12 mm and placed about 10 mm apart in the region of the lower premolars. The fixtures healed in a closed environment for 12 weeks, after which they were uncovered and loaded with abutments and orthodontic devices to produce horizontal distraction with a force of 2 N (about 204 gm) for 12 weeks. Subsequently they were loaded with 5 N (about 510 gm) for another 24 weeks. The distance between and the mobility of the implants were determined before and after each phase of experimental loading. Fixtures of the same type that were osseointegrated and not exposed, or osseointegrated and loaded by mastication, were used as a control. Animals were euthanized and specimens sectioned. The result was that continuously loaded implants showed no significant displacement for any force level. The mobility of the fixtures increased slightly by about 1 Periotest-value (PTV) at the end of the experiment. No significant peri-implant pocket could be seen in implants loaded by continuous or masticatory forces. Histologic and morphometric evaluation indicated compaction of bone as a result of loading. Osseointegrated implants have potential as a firm osseous anchorage for orthodontic treatment and can resist continuous horizontal forces of at least 5 N (about 510 gm) during a period of several months. PMID- 9743135 TI - Retention and stability: a review of the literature. AB - Long-term posttreatment stability is an issue of great concern to all orthodontists. This article highlights the factors reported to play a role in posttreatment crowding and reviews the long-term retention studies evaluating the stability of various treatment modalities. Recommendations, based on well documented basic principles, are made to try to insure greater posttreatment stability of our orthodontically treated cases. PMID- 9743136 TI - A significant transverse discrepancy: a case with a high mandibular plane angle, a severe maxillary arch length deficiency, and significant transverse discrepancy. AB - A case report presented to the American Board of Orthodontics in partial fulfillment of the requirements for certification. PMID- 9743137 TI - Correlation between cortical plate proximity and apical root resorption. AB - Root resorption is one of the most common iatrogenic sequelae of orthodontic treatment. Recently, root contact with the labial or palatal cortical plate at root apex level during orthodontic tooth movement was reported to be related to root resorption, and dentofacial morphology was suggested to predispose certain persons to root contact with the cortical plate. In this study, we constructed a best-fit straight line for the maxillary palatal cortical plate and set a line for the labial cortical plate from A point to Prosthion point in order to obtain measurements of proximity of root apices with the cortical plates of the maxillary alveolus. We investigated the correlation between apical root resorption and the measured variables. Our findings suggest that root approximation to the palatal cortical plate during orthodontic treatment could explain approximately 12% of the variance observed in the level of root resorption and the maxillary alveolar bone width about 2%. Tooth extrusion and crown lingualization also contributed to root resorption. We concluded that maxillary central incisor apical root resorption is influenced by root approximation to the palatal cortical plate during orthodontic treatment. PMID- 9743138 TI - Dental and skeletal changes after intraoral molar distalization with sectional jig assembly. AB - The present study was conducted on 10 subjects to evaluate dental and skeletal changes after intraoral molar distalization. The maxillary molars were distalized with a sectional jig assembly. Sentalloy open coil springs were used to exert 150 gm of force for a period of 12 weeks. A modified Nance appliance was the main source of anchorage. The pre- and postdistalization records included dental study casts, clinical photographs, and cephalograms. A total of 665 readings recorded from lateral cephalograms and dental casts were subjected to statistical analysis. The mean distal movement of the first molar was 2.78 mm, which was highly significant (o < 0.001). It moved distally at the rate of 0.86 mm/month. There was clinically some distal tipping (3.50 degrees) and distopalatal rotation (2.40 degrees). These changes were statistically significant (p < 0.001). The second molars accompanied the first molars and moved distally by nearly the same amount. There was 1.00 mm increase in the overjet and 2.60 degrees mesial tip of second premolar. The changes in the facial skeleton and dentition bases were minimal and statistically not significant. However, there was clockwise rotation of the mandible of 1.30 degrees that was statistically significant. This was the result of molar extrusion (1.60 mm). PMID- 9743139 TI - Correlations between condylar characteristics and facial morphology in Class II preadolescent patients. AB - The aim of this retrospective study was to determine correlations between condylar characteristics measured from preorthodontic tomograms of preadolescents and their facial morphologic characteristics. The sample consisted of 136 patients displaying a Class II malocclusion, a vertical or horizontal skeletal growth tendency, and ranging in age between 10 years 0 months and 12 years 6 months for males and 9 years 0 months and 11 years 6 months for females. Two groups were established: the vertical group had 68 patients, 36 males and 32 females, (average pretreatment age, 11 years 0 months); the horizontal group also had 68 patients, 29 males and 39 females, their average pretreatment age was 10 years 9 months. The central cut of axially corrected lateral tomograms of the left and right temporomandibular joints for each group was randomized, blinded, and traced for condyle/fossa measurements including: anterior, superior and posterior joint space; condylar head and posterior condylar ramus inclination; condylar neck width; and condylar shape and condylar surface area. A logistic discriminant analysis with significance values set at p < 0.05 was used to determine the most reliable condylar characteristics to predict facial morphology. A cluster analysis was completed on the significant variables to form three clusters. Numeric ranges separating these clusters were then calculated. Chi-square tests measures of association were computed for significant variables and tested for associations between facial morphologic characteristics. Condylar head inclination and superior joint space proved to be significantly correlated to facial morphology (p values ranged from 0.010 to 0.018). Patients with vertical facial morphologic characteristics displayed decreased superior joint spaces and posteriorly angled condyles. Increased superior joint spaces and anteriorly angled condyles were significantly correlated to patients with a horizontal facial morphology. No significant correlations between the other condylar characteristics and facial morphology were determined. PMID- 9743140 TI - Individual growth in Class III malocclusions and its relationship to the chin cap effects. AB - Individual growth characteristics of the maxilla and the mandible in Class III malocclusions were investigated in terms of growth amount, growth direction, and timing of growth; chin cap effects were considered in the context of growth characteristics of the jaws. Longitudinal cephalograms of six untreated Japanese Class III subjects were used for the analysis of jaw growth from 8 to 14 years of age. The facial patterns were classified into the five groups with different effects of therapy by discriminant functions derived from our previous chin cap study. The results obtained were as follows: (1) Among the six subjects, inhibition of maxillary forward growth was found in four subjects at ages before the maximum pubertal growth in body height occurred, resulting from occlusal interference of anterior cross bite. The growth potential of the jaws appeared genetically normal, and the facial patterns were classified into groups in which chin caps have been found to be effective to some extent. (2) A subject with growth characteristics of strong mandibular downward growth was classified in a group showing backward growth of the mandible by chin caps. (3) One of the remaining sample showing the strongest mandibular forward growth was classified in a group where chin caps are not effective. (4) It was concluded that effects of chin cap therapy are closely related to growth characteristics of the mandible in Class III treatment. PMID- 9743141 TI - How to make Internet searches easier: tips for effective use of Web search engines. PMID- 9743142 TI - Litigation, legislation, and ethics. An ethical dilemma. PMID- 9743143 TI - What constitutes success in cancer surgery? Measuring the value of specialist care. PMID- 9743144 TI - Tuberculosis control is a team sport. PMID- 9743145 TI - Diagnosing malignant pleural mesothelioma. PMID- 9743146 TI - Tissue capnometry as a monitoring strategy for critically ill patients: just about ready for prime time. PMID- 9743147 TI - Methodologic standards for the evaluation of diagnostic tests: the need to evaluate the standards. PMID- 9743148 TI - Weaning from mechanical ventilation: what have we learned and what do we still need to know? PMID- 9743149 TI - Specialists achieve better outcomes than generalists for lung cancer surgery. AB - OBJECTIVE: A push toward care provided by generalists as opposed to specialists has occurred in the health-care marketplace despite a lack of provider specific outcome data. The objective of this study was to determine whether the outcome of patients undergoing lung cancer surgery is different between general surgeons (GSs) and thoracic surgeons (TSs). DESIGN: Examination of data from a state-wide severity-adjusted administrative hospital discharge database. SETTING/PARTICIPANTS: Patients undergoing lung cancer resection in all nonfederal acute care hospitals within South Carolina. MAIN OUTCOME MEASURES: Mortality by specialty adjusted for case mix. RESULTS: From 1991 to 1995, 1,720 resections for lung cancer were performed in South Carolina. One hundred thirty-seven cases were excluded because surgeons did not meet the predefined criteria for board certification, leaving 1,583 resections for analysis. One-half of lobectomies and nearly 60% of pneumonectomies were performed by GSs. Patients were similar in age, sex, gender, race, and the proportion in each severity of illness subclass. Mortality was significantly higher in patients who underwent lobectomy by GSs vs TSs (5.3% vs 3.0%; p<0.05) and in patients with extreme comorbidities (43.6% vs 25.4%; p=0.03) or age >65 years (7.4% vs 3.5%; p<0.05). Seventy percent of TSs performed > 10 cases in the series, whereas 75% of GSs performed <10 (p=0.05). Logistic regression analysis failed to identify any significant variable that might explain the mortality differences between TSs and GSs. CONCLUSION: Mortality is lower for lung cancer resection when the surgery is performed by a TS. PMID- 9743150 TI - Screening by specialists to reduce unnecessary test ordering in patients evaluated for tuberculosis. AB - STUDY OBJECTIVE: To determine if screening by specialists could reduce unnecessary test ordering and reduce costs related to diagnostic workup in patients undergoing evaluation for tuberculosis. DESIGN: Prospective evaluation of expert opinion in consecutive patients suspected of having tuberculosis. SETTING: A large municipal hospital. PATIENTS: Patients for whom sputum acid-fast smears were ordered. INTERVENTION: For patients from whom sputum acid-fast bacilli smears and cultures were requested, the chest radiograph and a brief clinical history were presented separately to two pulmonologists with considerable experience in tuberculosis. Each expert reviewed each case independently (and was blinded to the opinion of the other) and indicated if he thought sputum smear examination and culture was, in fact, necessary. Final clinical diagnosis and microbiological information were correlated with the experts' opinion. MEASUREMENTS AND MAIN RESULTS: Ninety-seven patients had sputum smears ordered and had chest radiographs available for review. The two experts believed that sputum examination (smear and culture) was indicated in only 51.5% and 52.6% of cases, respectively. Interobserver agreement was 84.4%. Ultimately, six cases of active tuberculosis were diagnosed. Each expert detected all proven cases of tuberculosis, although one case occurred in a patient with a poor quality radiograph about which the experts offered no opinion. CONCLUSIONS: Screening by experienced clinicians may be effective in reducing unnecessary test ordering and reducing costs related to diagnostic workup in patients evaluated for tuberculosis. PMID- 9743151 TI - Relative occurrence of flow limitation and snoring during continuous positive airway pressure titration. AB - OBJECTIVES: To examine the relative temporal appearance of flow limitation and snoring during continuous positive airway pressure (CPAP) titration, compare their sensitivity as indicators of airway obstruction, and assess their relative utility as feedback variables for automatic titration of CPAP. DESIGN: Retrospective review of data. SETTING: University teaching hospital. PATIENTS: Fifty-three patients diagnosed as having obstructive sleep apnea or upper airway resistance syndrome undergoing CPAP titration. MEASUREMENTS AND RESULTS: We used a prototype automatic CPAP device that adjusts pressure in response to apnea, snoring, and/or flow limitation. The present study takes advantage of the frequent automatic decreases in pressure from a therapeutic level, as well as any technician-initiated decreases in pressure. We tabulated, for each pressure decrease of >0.4 cm H2O, the occurrences of snoring alone, flow limitation alone, or simultaneous appearance of both. Of 2,177 automatic pressure decreases, 64% resulted in flow limitation alone, 8% in snoring alone, and 22% in the simultaneous occurrence of both. Overall, 86% of decreases resulted in flow limitation alone or were simultaneous with snoring, whereas 30% of decreases resulted in snoring alone or were simultaneous with flow limitation. In 10 of 35 patients, snoring alone occurred in > 10% of the pressure decreases. In all but 5 of 133 manual pressure decreases, flow limitation developed at or above the pressure at which snoring developed. CONCLUSIONS: While detection of snoring occasionally provided additional information, overall flow limitation was the earliest indicator of obstruction during decreases in CPAP. PMID- 9743152 TI - Effects of nasal continuous positive airway pressure therapy on respiratory parameters of upper airway patency in patients with obstructive sleep apnea syndrome. AB - OBJECTIVES: To assess whether an initial treatment with nasal continuous positive airway pressure (NCPAP) therapy, applied for one night, had any effect on airway patency. METHODS: In 18 patients with obstructive sleep apnea syndrome (OSAS), we measured the total resistance of the respiratory system (Rrs) and their relevant lung functions before and after polysomnography, with and without NCPAP therapy. The Rrs was measured at 3 Hz with the forced oscillation technique. The overnight changes in the specific respiratory conductance (SGrs=reciprocal of the Rrs per unit lung volume) was also calculated in the sitting position. Since many reports have suggested that obesity, through fat deposits around the pharynx, can affect the mechanical and neuromuscular properties of the upper airway, we also investigated if the degree of obesity was related to the magnitude of improvement in these parameters. RESULTS: After the first night of NCPAP therapy, the Rrs decreased (sitting: 4.8+/-0.4 vs 4.3+/-0.4 cm H20/L/s, p < 0.05; lying: 6.5+/-0.4 vs 5.6+/-0.4 cm H20/L/s, p < 0.05) and the maximal voluntary ventilation increased in the morning (sitting: 101.6+/-5.8% vs 106.4+/-4.5%, p < 0.05; lying: 91.2+/-5.4% vs 97.9+/-4.7%, p < 0.05). The overnight difference in the SGrs showed a significant improvement after the initial treatment with NCPAP therapy (p < 0.05). However, the lung volume, flow volume loop, and closing volume in the morning did not change significantly after the therapy. An overnight decrease in the Rrs following NCPAP therapy is significantly correlated with the body mass index (sitting: r=0.54, p < 0.05; lying: r=0.61, p < 0.01). CONCLUSION: The improvements in Rrs without changes in spirometry may reflect improved upper airway patency after NCPAP therapy. The degree of obesity is suggested to be associated with the treatment effect on upper airway in patients with OSAS. PMID- 9743153 TI - Psychiatric symptoms in sleep apnea syndrome: effects of gender and respiratory disturbance index. AB - BACKGROUND: Previous studies have suggested an association between Sleep Apnea Syndrome (SAS) and several psychiatric disorders such as depression and anxiety. STUDY OBJECTIVE: To evaluate the association of SAS with psychiatric symptoms as determined by the SCL-90 psychiatric questionnaire. METHODS: The study comprised 2,271 patients (1,977 men, 294 women) referred to the Technion Sleep Laboratories with suspected SAS. They completed the SCL-90 Symptom Self-Report Inventory and then underwent a whole-night polysomnographic examination. The study population was stratified into subgroups according to gender, age, and respiratory disturbance index (RDI). RESULTS: Among men, there were no body mass index, RDI, or age-related differences in anxiety, depression, or in any other SCL-90 dimension. The depression and anxiety scores were significantly higher in women than in men for all age groups and for all levels of RDI. The depression score was higher in women with severe SAS than in women with mild SAS, for all ages. Surprisingly, in women who were only simple snorers, the depression and anxiety scores were higher than in mild SAS sufferers, for all age groups. CONCLUSIONS: In our large male population, neither the existence nor the severity of SAS was associated with depression or anxiety. Women had higher anxiety and depression scores, independent of other factors, than men. Women with severe SAS had higher depression scores than women with mild SAS. PMID- 9743154 TI - Percutaneous transthoracic needle aspiration biopsy: a comprehensive review of its current role in the diagnosis and treatment of lung tumors. AB - OBJECTIVE: The purpose of this study is to examine the accuracy and complications of transthoracic needle aspiration biopsy (TTNA) to determine its optimal role in the evaluation of patients with lung tumors. MATERIALS AND METHODS: The charts of 130 consecutive patients who had undergone CT-guided TTNA were reviewed retrospectively. Thirty-two (25%) of these patients had subsequent surgery and 5 had subsequent transbronchial biopsy (TBB). Using the final surgical and TBB diagnosis as a reference, the accuracy, sensitivity, specificity, and prevalence of malignancy were calculated. Each case was also examined to determine the presence or absence of complications. RESULTS: Of the 130 biopsy results, 95 (73%) were malignant, 33 (25%) were nonspecific, and only 2 (2%) had a specific benign diagnosis. Thirty-two patients subsequently underwent surgical resection. The overall prevalence of malignancy after surgical diagnosis was 91%. The overall diagnostic accuracy of TTNA was 76%. The sensitivity of TTNA for the detection of malignancy was 74% and its specificity was 100%. When comparing TTNA results of small (<3 cm) and large (> or = 3 cm) tumors, the occurrence of nonspecific results was 36% and 16%, respectively. Fifty-six (43%) patients had a pneumothorax subsequent to TTNA. Twenty-four (43%) of these patients required a chest tube and remained hospitalized for a mean of 6 days. CONCLUSION: Patients who are surgical candidates and have a high clinical suspicion for malignancy should undergo surgical biopsy and resection of their lung tumors if indicated. Information gained from TTNAs performed on this patient population will rarely result in a change in their clinical management. PMID- 9743155 TI - Primary peripheral lung carcinoma smaller than 1 cm in diameter. AB - BACKGROUND: Several investigators have reported on the risk of limited resection in patients with small peripheral lung cancer. Primary peripheral lung carcinomas 1 cm or less in maximum dimension were reviewed to study the feasibility of limited surgery. METHODS: Among 1,051 lung cancer patients who underwent surgical resection of the lung in the National Cancer Center Hospital East, Kashiwa, Chiba, Japan, from January 1986 through March 1997, there were 13 patients who had untreated peripheral cN0M0 tumors 1 cm or less in maximum dimension on the resected specimens who underwent systematic mediastinal dissection. Their specimens were histopathologically reviewed. RESULTS: There were ten adenocarcinomas, one small cell carcinoma, one poorly differentiated squamous cell carcinoma, and one typical carcinoid tumor. One adenocarcinoma showed lymphatic vessel invasion, venous invasion, and a subcarinal node metastasis. The small cell carcinoma was accompanied by a lymph node metastasis in a segmental node. The small cell carcinoma and another adenocarcinoma showed lymphatic vessel invasion. Of the ten adenocarcinomas, six were Noguchi's type B and four were type C. CONCLUSION: Even among the pulmonary peripheral cancers smaller than 1 cm in diameter, more than one third showed an invasive nature. This fact must be considered in selecting limited resection in these patients. It is evident that tumor size alone cannot be an indicator for limited resection in lung cancer patients. PMID- 9743156 TI - Metabolic imaging of malignant pleural mesothelioma with fluorodeoxyglucose positron emission tomography. AB - BACKGROUND: The diagnosis of malignant mesothelioma is a challenging medical problem. CT often cannot differentiate between benign diffuse pleural thickening and malignant mesothelioma, while thoracentesis and CT-guided biopsies are insensitive. We have assessed the value of positron emission tomography (PET) with 2-fluoro-2-deoxy-D-glucose (FDG) in the evaluation of malignant mesothelioma. METHODS: Twenty-eight consecutive patients referred for the evaluation of suspected malignant mesothelioma were evaluated by FDG-PET imaging. Measured attenuation correction was performed in 26 of 28 cases for quantitation with the standardized uptake value (SUV) method. The results of PET imaging were compared with those of video-assisted thoracoscopy or surgical biopsies. RESULTS: Surgical biopsy specimens confirmed the presence of malignant disease in 24 patients and demonstrated benign processes in the remaining four. The uptake of FDG was significantly higher in malignant than in benign lesions (SUV=4.9+/-2.9 and SUV=1.4+/-0.6, respectively; p<0.0001). With a SUV cutoff of 2.0 to differentiate between malignant and benign disease, a sensitivity of 91% and a specificity of 100% could be achieved, although the activity in some epithelial mesotheliomas tended to be close to this threshold. FDG-PET images provided excellent delineation of the active tumor sites. Hypermetabolic lymph node involvement was noted on FDG-PET images in 12 patients, 9 of which appeared normal on CT scans. Histologic examination in six patients confirmed malignant nodal disease in five cases and indicated granulomatous lymphadenitis in one. CONCLUSION: In this highly selected population, FDG-PET imaging was a sensitive method to identify malignant mesothelioma and determine the extent of the disease process. PMID- 9743157 TI - Complement components and their activation products in pleural fluid. AB - STUDY OBJECTIVES: The aim of this study was to determine the role of complement components in pleural effusion measured with novel markers of complement activation, to assess which pathway of activation is predominant in different diseases, and to find out whether the analysis of complement components and their activation products could help in diagnostic procedure differentiating the etiologies of pleural effusion. PATIENTS: The study population consisted of 71 patients who had pleural effusion secondary to tuberculosis (n=23), rheumatic disease (n=10), or malignancy (n=38). MEASUREMENTS: Complement components and their activation products, including the soluble terminal complex SC5b-9, were measured in plasma and pleural fluid. RESULTS: In all patients with rheumatic pleurisy, pleural fluid SC5b-9 was higher than 2 AU/mL and in all patients with malignant pleural fluid it was lower than 2 AU/mL. The mean level of SC5b-9 in rheumatic pleural effusion was also significantly higher than in tuberculosis. In addition, the concentrations of pleural fluid C3 and C4 were significantly lower and the ratio C4d/C4 was significantly higher in rheumatic compared with tuberculous or malignant pleurisy. In plasma, both SC5b-9 and C1s-C1r-C1INH complexes were significantly higher in rheumatic subjects than in other patients. In stepwise multinominal logistic regression analyses, the most significant predictors for rheumatic pleural fluid were high pleural fluid SC5b-9 and low C4. CONCLUSIONS: These observations indicate that the complement cascade is activated through both the classic and the alternative pathways in rheumatic pleurisy. Determinations of SC5b-9 and C4d/C4 in pleural fluid were the best variables differentiating rheumatic, tuberculous, and malignant effusions. PMID- 9743158 TI - Medical thoracoscopic talc pleurodesis for chylothorax due to lymphoma: a case series. AB - STUDY OBJECTIVES: Recurrent chylothorax as a complication of lymphoma has had unsatisfactory outcomes. Serial thoracentesis, tube thoracostomy, and pleurodesis via chest tube have been ineffective and compromise the nutritional and immune status of the patient. Medical thoracoscopic talc pleurodesis has been safe and effective in the treatment of some other varieties of recurrent pleural effusions. Our objective was to investigate the safety and efficacy of medical thoracoscopic talc pleurodesis in the palliation of chylothorax related to lymphoma. DESIGN: This is a report of 24 hemithoraces treated in 19 consecutive patients with lymphoma-related chylothorax, failing chemotherapy or radiation therapy. The average patient age was 55 years. INTERVENTIONS: Medical thoracoscopy was performed under local anesthesia and conscious sedation in a bronchoscopy suite. Sedation included midazolam (mean dose, 6 mg; range, 2-14 mg) with either meperidine (mean dose, 94 mg; range 25-140 mg), or morphine (mean dose, 18 mg; range 4-40 mg). Pleurodesis was performed with insufflation of sterile asbestos-free talc, (4-8 g). After pleurodesis, chest tubes were placed, with the mean duration of chest tube placement being 4 days, range 3 to 10 days. RESULTS: One patient died a few days after the procedure due to causes related to the primary disease process. Follow-up was for at least 90 days following the procedure. Patients were assessed at 30, 60, and 90 days following the procedure. At each of these endpoints, all patients remaining alive were without recurrence of pleural effusions, which was confirmed by chest radiography. Eight patients in the series died of the effects of their malignancy during the 90-day evaluation interval. Complications included medication reactions in two patients (8.3%) and ARDS in one patient (4.1%). CONCLUSION: Many patients with lymphoma-related chylothorax are refractory to chemotherapy and/or radiation therapy. In this group, medical thoracoscopic talc pleurodesis has an acceptable complication rate and a 100% success rate in the prevention of recurrence of pleural effusions at 30, 60, and 90 days following the procedure. PMID- 9743159 TI - Response to symptom-limited exercise in patients with the hepatopulmonary syndrome. AB - OBJECTIVE: To study the response to symptom-limited exercise in patients with the hepatopulmonary syndrome (HPS). DESIGN: The response to maximal cardiopulmonary exercise (CPX) was studied in 5 patients with HPS and compared with 10 case control (normoxemic, NC) cirrhotics (matched for age, gender, etiology and severity of liver disease, tobacco use, and beta-blocker therapy) and 9 hypoxemic control cirrhotics (HC) without clinical evidence of HPS. SETTING: Cardiopulmonary exercise physiology laboratory in a tertiary care referral center. PATIENTS: Cirrhotics referred for CPX as part of their preliver transplantation evaluation. MEASUREMENTS: Standard pulmonary function tests and echocardiography were performed to assess resting pulmonary and cardiac function. Peak oxygen consumption (VO2), minute ventilation, arterial blood gases, and dead space (VD/VT) were determined during symptom-limited maximal CPX. RESULTS: Resting spirometry and lung volumes were similar between HPS and NC subjects, while HC subjects had restrictive physiology. Differences existed in diffusing capacity corrected for hemoglobin and alveolar volume percent predicted (HPS, 45+/-2 vs NC, 68+/-3, p<0.05; vs HC, 70+/-4, p<0.05), PaO2 (HPS, 70+/-5 mm Hg; HC, 79+/-3 mm Hg, vs NC, 102+/-3 mm Hg, p<0.05) and alveolar-arterial (A-a) O2 gradient (HPS, 42+/-8 mm Hg vs HC, 27+/-2 mm Hg, p<0.05; vs NC, 6+/-2 mm Hg, p<0.05). During CPX, HPS patients achieved a lower peak VO2 percent predicted (HPS, 55+/-6 vs NC, 73+/-3, p<0.05; vs HC, 71+/-5, p<0.05) and VO2 at the ventilatory threshold as percent predicted peak VO2 (HPS, 36+/-2 vs NC, 55+/-4, p<0.05; vs HC 55+/-5, p<0.05). While no differences existed in heart rate and breathing reserve, HPS patients had significantly lower PaO2 (HPS, 50+/-5 mm Hg vs NC, 97+/-4 mm Hg, p<0.05; vs HC, 87+/-6 mm Hg, p<0.05), wider A-a O2 gradient (HPS, 73+/-5 mm Hg vs NC, 13+/-3 mm Hg, p<0.05; vs HC, 31+/-5 mm Hg, p<0.05) and higher VD/VT (HPS, 0.36+/-.03 vs NC, 0.18+/-.02, p<0.05; vs HC, 0.28+/-.02, p<0.05) at peak exercise. For HPS patients, VO2 was negatively correlated with VD/VT (r2=0.9) and positively correlated with PaO2 (r2=0.41) at peak exercise. CONCLUSIONS: Patients with HPS demonstrate a severe reduction in aerobic capacity, beyond that found in cirrhotics without syndrome. The significant hypoxemia and elevated VD/VT at peak exercise suggest that an abnormal pulmonary circulation contributes to further exercise limitation in patients with HPS. PMID- 9743160 TI - Effects of long-term aerosol pentamidine for Pneumocystis carinii prophylaxis on pulmonary function. AB - OBJECTIVE: The purpose of the study was to evaluate the long-term effects of aerosolized pentamidine (AP) on the pulmonary function of HIV-positive individuals receiving AP for Pneumocystis carinii pneumonia (PCP) prophylaxis. DESIGN: A retrospective analysis of serial pulmonary function tests (PFTs) performed on a cohort of HIV-positive patients who had received AP for >2 years. METHODS: Of the 1,787 HIV-positive patients receiving AP prophylaxis in our database, 380 patients had been receiving AP for at least 24 months. Of these 380 patients, the PFTs at baseline and at 24 months after starting AP therapy were documented in 179. We compared the baseline PFTs of these 179 patients with results obtained 24 months later to evaluate if there was any change in pulmonary function parameters. RESULTS: Baseline and 24-month PFT parameters (total lung capacity [TLC], FVC, residual volume [RV], FEV1, FEV1/FVC, maximum midexpiratory flow rate at 50% of vital capacity [MEF50], diffusion of carbon monoxide [DLCO]) were all within the normal ranges. However, AP therapy over 24 months was associated with a modest decline in lung volume parameters (TLC, FVC, RV), a slight reduction in flow rates (FEV1, FEV1/FVC, and MEF50), but no change in DLCO. The mean treatment duration was 23.8 months (range, 21 to 27 months). CONCLUSIONS: Long-term AP therapy is associated with a mild combined restrictive and obstructive pulmonary defect. Since 24-month PFT parameters remained within the normal ranges, this level of decline is of unclear clinical significance. Overall, AP has good pulmonary tolerance when used up to 24 months for PCP prophylaxis in patients with normal baseline pulmonary function. PMID- 9743161 TI - The safety of brachial artery puncture for arterial blood sampling. AB - OBJECTIVE: This study was designed to determine the incidence of complications in a sample of 6,185 brachial artery punctures for arterial blood gas analysis. METHODS: The study sample was comprised of adult patients who had arterial blood gas analysis ordered in the course of their clinical evaluations in a multispecialty clinic and hospital affiliated with a university school of medicine. Subjects were entered prospectively at the time the procedure was done. RESULTS: The overall incidence of all complications was 2.0%. Immediate limb pain or parenthesias occurred in 1.1%, while the onset of symptoms was delayed up to 24 h in 0.9%. Hematoma formation occurred in only 0.06%. None of the complications was considered to be of major impact, in that none was associated with limb ischemia or other objective abnormalities. Only one subject required analgesic medication to control pain that ultimately subsided spontaneously without deficit. CONCLUSION: We believe that brachial artery puncture, when properly performed, is a safe and reliable alternative route for obtaining arterial blood for gas analysis. PMID- 9743162 TI - Effect of exposure to low levels of ozone on the response to inhaled allergen in allergic asthmatic patients. AB - STUDY OBJECTIVES: In a previous study published by our group, six out of nine subjects with mild allergic asthma were shown to have an enhanced response to allergen challenge following a 1-h exposure in an 0.8-m3 exposure chamber (modified from a body plethysmograph) to an average of 120 parts per billion (ppb) ozone at rest. Other studies failed to confirm this effect. In the present study, using a similar design, we reexamined this effect using a larger group of asthmatics and a larger chamber allowing minimal fluctuations in ozone levels during exposures. DESIGN: Prospective, randomized single-blinded crossover study. SETTING: Pulmonary function laboratory equipped with an exposure chamber. SUBJECTS: Fifteen subjects had mild allergic asthma; 9 men and 6 women; the mean (SD) age was 32.5 (10) years; FEV1 was 3.4 (0.8) L; baseline methacholine provocation concentration causing a 20% fall in FEV1 was (PC20) 3.28 (4.1) mg/mL. INTERVENTIONS: Each participant was exposed, at rest, on 1 day to filtered air and on another day to ozone (mean level=120 ppb) in a larger exposure chamber than the one used in our first study with less variability in ozone level (110 to 130 vs 85 to 175 ppb) using a random, single-blinded design. After each exposure, the subject was challenged with allergen (nine with grass pollen extract and six with ragweed extract) and allergen PC15 was measured. RESULTS: Ozone preexposure did not affect allergen PC15 when compared with clean air preexposure (allergen PC15 dilution 1/114 vs 1/119, respectively). Ozone vs air preexposure resulted in an allergen PC15 that was lower in five subjects, higher in six, and unchanged (within one doubling dose) in four. CONCLUSIONS: At this low level with less variability and lower peaks than our previous study, ozone had no significant effect on airway allergen responsiveness. PMID- 9743163 TI - A comparison of triamcinolone acetonide MDI with a built-in tube extender and beclomethasone dipropionate MDI in adult asthmatics. AB - STUDY OBJECTIVE: In this study, the efficacy and safety of triamcinolone acetonide (TA) metered-dose inhaler with a built-in tube extender and beclomethasone dipropionate (BDP) metered-dose inhaler without a spacer device were compared. Both treatments were dosed at their most commonly used daily doses (within labeling). DESIGN: A 56-day, randomized, double-blind, double-dummy, placebo-controlled trial. SETTING: Seventeen asthma/allergy centers. PATIENTS: We enrolled 339 patients 18 to 65 years of age, with a documented history of bronchial asthma (FEV1, 50 to 90% of predicted value) for > or = 2 years who required inhaled corticosteroid therapy. INTERVENTIONS: Patients were randomized to receive BDP 336 microg/d (4 puffs bid) plus TA placebo (4 puffs bid), TA 800 microg/d (4 puffs bid) plus BDP placebo (4 puffs bid), or TA and BDP placebos (4 puffs of each bid). The only other asthma medication permitted was inhaled albuterol that was used as a rescue medication. All medications were administered via the closed-mouth inhalation technique. MEASUREMENTS AND RESULTS: At 8 weeks and at study end point, both active treatment groups had statistically significant and comparable improvements in FEV1 relative to baseline, and statistically significant increases relative to placebo. At study end point, improvements in forced expiratory flow (FEF25.75%), clinic peak expiratory flow (PEFR), and FVC were statistically significant for the active treatment groups compared with placebo. At end point, the mean difference between BDP and TA for mean change in FEV1 from baseline in the efficacy population was 0.02 and the 95% confidence interval was -0.11, 0.15. Asthma symptoms recorded at clinic visits showed statistically significant improvements for the BDP and TA groups compared with the placebo group. Treatment-related adverse events occurred with similar frequency in all patient groups-25.5% of placebo-treated patients, 22.3% of BDP patients, and 20.4% of TA patients. The incidence of oropharyngeal adverse events, including cough, thrush, and dysphonia, was not statistically different between the two active treatment groups. CONCLUSION: In this randomized, double blind, placebo-controlled study of adult asthmatics treated with either BDP without a spacer or TA with its built-in tube extender, BDP and TA were comparable in efficacy as measured by FEV1 and other pulmonary function tests, and by improvement in asthma symptoms. Both active treatments were significantly more effective than placebo. All treatment groups were comparable in safety as measured by the incidence of adverse events. PMID- 9743164 TI - Comparison of the Maxair Autohaler to wet nebulizer in patients with acute asthma. AB - STUDY OBJECTIVE: Patients with acute asthma often have difficulty using a conventional metered-dose inhaler. The Maxair Autohaler (3M Pharmaceuticals; St. Paul, MN) is a hand-held breath-actuated device developed to help patients coordinate drug administration. The study objective is to compare the efficacy of the Autohaler with inhaled beta-agonist administered by wet nebulizer in treating acute asthma exacerbations. DESIGN: Parallel, randomized, placebo-controlled clinical trial. SETTING: Emergency department (ED) of a university-affiliated hospital. PARTICIPANTS: Patients aged 18 to 55 years presenting to the ED with acute asthma and an FEV1 40 to 70% on hospital arrival. INTERVENTIONS AND MEASUREMENTS: Patients were given either six puffs of the Maxair Autohaler (1,200 microg pirbuterol) plus saline solution by nebulizer (active Autohaler group) or six puffs placebo by Autohaler plus 2,500 microg albuterol via nebulizer (active nebulizer group). Treatment was repeated at 30 and 60 min, with clinical evaluation performed before each treatment and again at 120 min. At 120 min, the protocol was completed. RESULTS: Twenty-four patients were enrolled, with 5 excluded because of protocol violations. Baseline FEV1 percent predicted values for the active Autohaler group were 53% vs 51% for the active nebulizer group (p=not significant [NS]). At time 120 min, values were 66% for Autohaler and 64% for nebulizer (p=NS). The average time to administer Autohaler was 2.9 min, vs 9.1 min for nebulizer. No patient was excluded because of the inability to use the Autohaler device adequately. CONCLUSION: In patients with moderate asthma exacerbations, similar improvements in pulmonary function are obtained when beta agonists are given by either the Maxair Autohaler or a wet nebulizer device. PMID- 9743165 TI - Pulmonary and aortic blood flow measurements in normal subjects and patients after single lung transplantation at 0.5 T using velocity encoded cine MRI. AB - PURPOSE: It is the purpose of this study to compare pulmonary and aortic blood flow measurements obtained in patients after single lung transplantation (SLTX) with those in volunteers. METHODS/MATERIAL: In nine patients after SLTX (three male, six female) and nine volunteers (seven male, two female), double oblique phase contrast cine-MRI sequences perpendicular to the direction of blood flow were obtained in the ascending aorta, main, right, and left pulmonary artery on a 0.5-T unit (Philips Gyroscan; Best, the Netherlands) (repetition time, 600 to 800 ms; echo time, 8 ms; alpha=30; field of view=280 mm matrix, 128x256, ECG gating, temporal resolution 16 time frames/RR interval). An initial in vitro study using the same sequence on a nonpulsatile flow phantom showed excellent correlation (r=0.99) between MRI measurements of flow velocity and flow volume and true velocity and flow volume. Measurements of blood flow volume (mL/min), peak mean systolic velocity, resistive index, and distensibility index were obtained in each vessel. RESULTS: We found excellent correlations between left and right cardiac output as measured by velocity encoded cine-MRI (VEC-MRI) in the ascending aorta and main pulmonary artery both in normal volunteers (r=0.95) and in patients (r=0.91). Differential pulmonary blood flow measurements in volunteers showed that 55% of the right cardiac output was directed to the right and 45% to the left lung. Differential pulmonary blood flow in patients showed that most of the blood flow (81%) reaches the transplanted lung and only 19% reaches the patient's own lung (SLTX: 4.5+/-1.8 L/min, patient's own lung: 1.2+/ 0.8 L/min). There were significant differences (p<0.05) in peak mean systolic velocity and resistive index obtained in the pulmonary arteries, both between normal volunteers and patients and between measurements obtained in the patient's own lung and the transplanted lung. CONCLUSION: VEC-MRI blood flow measurements are a promising noninvasive tool to monitor the hemodynamic changes of pulmonary blood flow after SLTX. PMID- 9743166 TI - Comparison of inhaled nitric oxide and inhaled aerosolized prostacyclin in the evaluation of heart transplant candidates with elevated pulmonary vascular resistance. AB - STUDY OBJECTIVE: Elevated pulmonary vascular resistance is a risk factor in heart transplantation and reversibility of high pulmonary vascular resistance is evaluated preoperatively in potential recipients using i.v. vasodilators or inhaled nitric oxide. Prostacyclin is a potent vasodilator, which when inhaled, has selective pulmonary vasodilatory properties. The aim of this study was to compare the central hemodynamic effects of inhaled prostacyclin with those of inhaled nitric oxide in heart transplant candidates. DESIGN: A pharmacodynamic comparative study. SETTING: Cardiothoracic ICU or laboratory for diagnostic heart catheterization at a university hospital. PATIENTS: Ten heart transplant candidates with elevated pulmonary vascular resistance (>200 dynes x s x cm(-5) and/or a transpulmonary pressure gradient > 10 mm Hg) were included in the study. INTERVENTIONS: Nitric oxide (40 ppm) and aerosolized prostacyclin (10 microg/mL) were administered by inhalation in two subsequent 10-min periods. Hemodynamic measurements preceded and followed inhalation of each agent. MEASUREMENTS AND RESULTS: Both inhaled nitric oxide and inhaled prostacyclin reduced mean pulmonary artery pressure (-7% vs -7%), pulmonary vascular resistance (-43% vs 49%), and the transpulmonary gradient (-44% vs -38%). With inhaled prostacyclin, an 11% increase in cardiac output was observed. Other hemodynamic variables, including the systemic BP, remained unaffected by each of the agents. CONCLUSIONS: Inhaled prostacyclin induces a selective pulmonary vasodilation that is comparable to the effect of inhaled nitric oxide. Major advantages with inhaled prostacyclin are its lack of toxic reactions and easy administration as compared with the potentially toxic nitric oxide requiring more complicated delivery systems. PMID- 9743167 TI - The short-term effects of digoxin in patients with right ventricular dysfunction from pulmonary hypertension. AB - OBJECTIVE: Studies on the effects of digoxin in patients with right ventricular failure and normal left ventricular function have not been performed. We evaluated the short-term effects of digoxin administration in patients with primary pulmonary hypertension on hemodynamics, neurohormones, and baroreceptor responsiveness. DESIGN: This was a prospective study with patients serving as their own controls. SETTING: University Hospital Intensive Care Unit with central monitoring. PATIENTS: Seventeen patients with primary pulmonary hypertension and symptomatic heart failure were enrolled. INTERVENTIONS: Following baseline hemodynamics, neurohormonal samples were drawn and the heart rate response to change in blood pressure following a challenge of phenylephrine and nitroprusside were recorded. One mg of intravenous digoxin was given and the measurements repeated after 2 hours. RESULTS: Following digoxin there was a significant increase in cardiac output (3.49+/-1.2 to 3.81+/-1.2 L/min., p=0.028), a significant fall in norepinephrine (680+/-89 to 580+/-85 pg/ml, p=.013), and a significant increase in atrial natriuretic peptide (311+/-44 to 421+/-9 pg/ml, p=0.01). All of the patients had changes in heart rate and blood pressure following phenylephrine and nitroprusside challenge, but there was no significant difference in the change in heart rate response to change in blood pressure when rechallenged after digoxin treatment. CONCLUSION: Digoxin produces a modest increase in cardiac output in patients with pulmonary hypertension and right ventricular failure, as well as a significant reduction in circulating norepinephrine. No detectable effects of digoxin on baroreceptor responsiveness were apparent. The use of digoxin in pulmonary hypertension is warranted. PMID- 9743168 TI - Quality of history taking in patients with aortic dissection. AB - STUDY OBJECTIVES: Aortic dissection generally is an acute catastrophe. Rapid diagnosis is critical. We hypothesized that the quality of history taking contributes to the accuracy of diagnosis in patients with dissection. DESIGN: Retrospective chart review of 83 patients, whose diagnosis of aortic dissection was confirmed by autopsy, surgery, CT scan, echocardiogram, or angiogram. The quality of the initial history was reviewed using predetermined criteria. The physicians' initial clinical impressions were recorded. RESULTS: The examining physician correctly suspected aortic dissection after the initial clinical evaluation in 54 of 83 patients (65%). Only 33 of 78 patients with symptoms (42%) were asked about the quality, location, and onset of their pain, the three descriptors identified a priori as important. In 19 patients (24%), only zero or one descriptor was recorded. When all three questions were asked, dissection was suspected in 30 of 33 patients (91%); when zero, one, or two questions were asked, dissection was suspected in 22 of 45 patients (49%). CONCLUSION: Despite important advances in diagnostic imaging, accurate diagnosis of aortic dissection requires an accurate history. In our series, the quality of initial history was associated with the accuracy of the initial clinical impression in patients with aortic dissection. PMID- 9743169 TI - Long-term results of fiberoptic bronchoscopic balloon dilation in the management of benign tracheobronchial stenosis. AB - STUDY OBJECTIVES: To assess the short- and long-term effects of balloon dilatation using a fiberoptic bronchoscope in the management of benign tracheobronchial stenosis (TBS). Treatment strategies have included open surgical resection or endoscopic techniques. The endoscopic techniques have included Nd:Yag laser, cryotherapy, stent placement, rigid bronchoscopy, and balloon dilation (BD). DESIGN: Prospective sequentiality study. SETTING: Tertiary care academic hospital. PATIENTS: This study consisted of 14 patients, ages ranging from 35 to 72 years, whose symptoms of dyspnea, cough, or wheeze were attributable to a TBS. INTERVENTION: This study describes the use of flexible fiberoptic bronchoscopy (FFB) with a balloon catheter for airway dilation. Each patient underwent FFB, in which a balloon catheter was threaded over the guidewire and positioned across the stenosis. Under direct visualization, the balloon was inflated for 30 to 120 s. Repeat inflation-deflation cycles were done if airway narrowing remained after the initial attempt. RESULTS: Seven patients had TBS following lung transplantation, three after sleeve resection, two following irradiation therapy, and two due to fibrosing mediastinitis. All patients had initial success: increased airway dimensions and symptom relief. Ten patients had successful airway dilation after one session. Of the four patients who required multiple sessions, one had cryotherapy and in each of the others an airway stent was placed. CONCLUSIONS: BD offers immediate symptom relief and can be used in conjunction with Nd:Yag laser, cryotherapy, stent placement, or mechanical dilation with a rigid bronchoscope. The advantages of FFB with BD include operator familiarity, patient safety, and avoidance of general anesthesia. PMID- 9743170 TI - African-American race and antibodies to topoisomerase I are associated with increased severity of scleroderma lung disease. AB - STUDY OBJECTIVES: To determine whether African-American race is independently associated with lung disease in scleroderma. DESIGN: Retrospective review. SETTING: University medical center in Baltimore. PATIENTS: One hundred one patients with diffuse cutaneous scleroderma with available serum samples. MEASUREMENTS: Patients underwent lung function testing as part of their routine clinical care. Percent predicted values adjusted for race were calculated for FVC, single-breath carbon monoxide diffusing capacity (Dco), and FEV1. Serum samples were assayed for the presence of antibodies to topoisomerase I and RNA polymerase II. RESULTS: Scleroderma patients of African-American race had lower percent predicted values than white patients for FVC (p<0.002), Dco (p<0.0001), and FEV1 (p<0.0001). Antibodies to topoisomerase I but not antibodies to RNA polymerase II were also associated with lung function. African-American scleroderma patients were distinct from white patients in having younger age of onset and higher prevalence of antibodies to topoisomerase I. In multivariate analyses accounting for sex, age, smoking history, years of scleroderma symptoms, and RNA polymerase II antibody status, African-American race and topoisomerase I antibody status independently predicted lower lung function. CONCLUSION: African American race and antibodies to topoisomerase I are independent risk factors for scleroderma lung disease. PMID- 9743171 TI - Acute Q fever pneumonia: a review of 80 hospitalized patients. AB - STUDY OBJECTIVES: To emphasize epidemiologic, clinical, or radiologic characteristics whose detection could lead to an early diagnosis and to enhance therapeutic efficacy. PATIENTS: Eighty hospitalized patients from 1982 to 1996. DESIGN: The diagnosis of Q fever infection was serologically confirmed in all the patients (phase II Coxiella burnetii antibody) using the complement fixation test and/or the indirect immunofluorescence antibody test. RESULTS: Patients from rural and urban areas were noted in the same proportion; however, the usual epidemiologic factors such as contact with cats or farm animals were found in 40% of the patients. Mean age+/-SD was 49+/-20 years, and there was a higher sex ratio of male to female patients (1:3.44). We found a specific seasonal distribution since 80% of the cases occurred between February and May. Delay before referring to hospital was 8.2+/-7.8 days, while 69.3% of the patients received an antibiotic treatment that was mainly penicillin or cephalosporin. The dominant clinical features were dry cough and high fever, as the maximal temperature reached more then 40 degrees C in 58% of the patients. Digestive symptoms were rare. WBC count remained within normal range in 80% of the cases with a low proportion of lymphocytes in half of the patients, and the sedimentation rate was usually elevated (55+/-34 mm). Altered liver function consisted more frequently in an elevated level of alkaline phosphatase (70% of the cases) than transaminases, while hyponatremia was frequently mentioned (28.2% of the patients). We found radiologic evidence of unique lobar or segmental alveolar opacity involving more likely the lower lobes in 55 patients, and multiple or interstitial opacities in the others. Chest radiographs were considered normal in eight patients. The clinical response was favorable in all the patients with a reduction in fever 4.8+/-3.9 days after the start of treatment with the second antibiotic that included mainly erythromycin or quinolones, and chest radiographs returned to normal in 81% of the patients within the first month. PMID- 9743172 TI - Increased iron and ferritin content of sputum from patients with cystic fibrosis or chronic bronchitis. AB - PURPOSE: Extracellular free iron, or iron bound to ferritin, may promote oxidative injury and bacterial growth in airways of patients with chronic airway inflammation due to cystic fibrosis (CF) or chronic bronchitis (CB). In this study, we assessed sputum content of total iron, ferritin, and transferrin in patients with CF or CB as well as sputum from normal subjects with acute airway inflammation caused by viral upper respiratory tract infections (URTIs). METHODS: Spontaneously produced sputum was obtained from 33 subjects, including 10 subjects with CF, 18 subjects with CB (10 acute exacerbations, 8 with stable CB), and 5 subjects with URTIs (control subjects). After lysing and dilution, total iron concentrations were determined by controlled coulometry, ferritin was measured by radioimmunoassay, and transferrin was measured by enzyme-linked immunosorbent assay. RESULTS: Iron was not present in detectable amounts in control sputums, but ferritin was present (6+/-2 ng/mg protein, mean+/-SE), as was transferrin (2.37+/-0.44 microg/mg). Compared with control subjects, concentrations of iron in sputum were increased in patient groups with higher amounts in CF patients (242+/-47 ng/mg, p<0.01) than CB patients with acute exacerbations or patients with stable CB (98+/-50 and 42+/-12 ng/mg, p<0.05 for both). Ferritin content of sputum was also increased in each group, with CF patients (113+/-22 ng/mg, p<0.001) higher than CB patients (acute, 45+/-10 ng/mg; stable, 87+/-24 ng/mg; p<0.01 for both). Compared with control subjects, sputum transferrin was decreased in CF patients (1.09+/-0.40 microg/mg, p<0.05), but not CB patients. CONCLUSIONS: These findings indicate there are increased airway concentrations of total iron and ferritin-bound iron in patients with CB and, to a greater extent, in patients with CF. Particularly in CF patients who also demonstrated decreased airway concentrations of transferrin, ferritin-bound iron in airways may promote oxidative injury and enhance bacterial growth. PMID- 9743173 TI - MRI of central venous anatomy: implications for central venous catheter insertion. AB - STUDY OBJECTIVES: To determine normative values for superior vena cava (SVC) length and the utility of radiographic landmarks for identifying the boundaries of the SVC for assisting central line placement. DESIGN: Cross-sectional study. SETTING: Urban tertiary care medical centers. PATIENTS: Patients undergoing thoracic MRI scanning for various indications. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: The SVC dimensions and relationship to radiographic landmarks were determined from MRI scans of 42 patients (22 men, 20 women; median age, 57 years). The median length of the SVC was 6.8 cm (range, 4.4 to 10.0 cm) and did not correlate with gender or other measured cardiovascular dimensions. The right tracheobronchial angle was the best radiographic landmark for determining the cephalad origin of the SVC being always caudad and within a median of 1.5 cm (range, 0.1 to 3.8 cm) of the upper SVC. It was always at least 2.9 cm above the atriocaval junction. The right superior heart border was formed by the left atrium in 38% (95% confidence interval, 23 to 53%) of patients and did not reliably identify the atriocaval junction. CONCLUSIONS: The right tracheobronchial angle is the most reliable landmark for the upper margin of the SVC. Venous catheters placed caudad to this landmark and cephalad to the right superior cardiac silhouette or no more than 2.9 cm caudad to the tracheobronchial angle result in catheter tips within the SVC. PMID- 9743174 TI - Efficacy of inhaled nitric oxide in children with ARDS. AB - STUDY OBJECTIVE: Data concerning inhaled nitric oxide (iNO) on pediatric ARDS is rare. We investigated the effects of iNO on pediatric ARDS in order to examine the ability to predict a response to iNO, the optimal concentration of iNO, the effects of < or = 1 ppm nitric oxide (NO), and the effect of iNO on PaCO2. SETTING: ICU at Kumamoto (Japan) University Hospital. PATIENTS AND INTERVENTIONS: Seven children with ARDS. The initial responses to 16 ppm NO and the dose response effects of 0.13 to 16 ppm NO were assessed. MEASUREMENTS AND RESULTS: Sixteen ppm of iNO improved oxygenation in all seven children. The use of iNO significantly increased the ratio of arterial oxygen tension to the fraction of inspired oxygen (PaO2/FIO2). A correlation between the NO-induced increase in PaO2/FIO2 and the baseline PaO2/FIO2 was observed (r=0.93, p<0.01). Dose-response tests showed that the optimal concentration of iNO was < or = 4 ppm, improvements in PaO2/FIO2 could be observed with concentrations of < or = 1 ppm NO, and iNO induced a slight decrease in PaCO2. CONCLUSIONS: In children with ARDS, iNO frequently improves oxygenation and induces a slight decrease in PaCO2, with the baseline PaO2/FIO2 functioning as a predictor of all NO response. Improvements of PaO2 and PaCO2 were observed with concentrations of iNO of < or = 1 ppm, a level in which the risk of a toxic reaction in children is minimal. Effects on outcome need verification in larger controlled trials. PMID- 9743175 TI - The physiologic effects of inverse ratio ventilation. AB - STUDY OBJECTIVES: The efficacy of inverse ratio ventilation in ARDS is not clear. Furthermore, the mechanism responsible has not been determined. We designed an animal study to determine if inverse ratio ventilation improves gas exchange and by what mechanism. DESIGN: Prospective randomized, controlled design was used. SETTING: University of Missouri Pulmonary/Critical Care Animal Laboratory. PARTICIPANTS: Nine dogs with oleic acid-induced lung injury as control animals to assess stability of the model, nine in the experimental model. INTERVENTIONS: Conventional ventilation with full recruitment extrinsic positive end-expiratory pressure (PEEP) was compared with two other modes of ventilation. One was inverse ratio with extrinsic PEEP and the second was inverse ratio with intrinsic PEEP equal to full recruitment PEEP. Full recruitment levels of PEEP were defined by optimizing compliance, then increasing PEEP by 2.5 cm/H2O. Each type of ventilation was maintained for 45 min after the edema had stabilized. Comparison of lung injury over time requires stability of the model over time. Therefore, we also assessed the stability of the preparation over time by examining compliance, extravascular lung water, and venous admixture in nine control dogs with equivalent lung injury over the same time span. MEASUREMENTS AND RESULTS: Mean airway pressure was increased by both types of inverse ratio ventilation, while compliance remained stable. Venous admixture was reduced (conv=0.32+/-0.12, inverse ratio with extrinsic PEEP=0.24+/-0.10, inverse ratio with intrinsic PEEP=0.28+/-0.11) with inverse ratio with extrinsic PEEP, but the improvement was less with inverse ratio with intrinsic PEEP, even though the mean airway pressure was higher. CONCLUSIONS: We conclude that increasing mean airway pressure by prolongation of the inspiratory time improves gas exchange in our model of ARDS, but when mean airway pressure is increased further, allowing the development of intrinsic PEEP, the beneficial effects on gas exchange are less. Increasing mean airway pressure with intrinsic PEEP is not equivalent to other methods of increasing mean airway pressure. PMID- 9743176 TI - Regulation of integrin-mediated adhesion by muscarinic acetylcholine receptors and protein kinase C in small cell lung carcinoma. AB - STUDY OBJECTIVES: Improved understanding of the phenotypic characteristics of small cell lung cancer (SCLC) cells may facilitate the development of new therapies for this bronchogenic malignancy with early metastases. Herein we investigate whether activation of the M3 subtype of muscarinic acetylcholine receptor (mAChR) expressed on SCLC cells affects beta1-integrin-mediated adhesion of these cells. DESIGN: Adhesion of the SCLC cell lines SCC-9 and NCI-H345 to extracellular matrix (ECM) proteins was investigated. Cell adhesion was quantified by labeling the cells with either toluidine blue dye and measuring optical density or 3H-thymidine and measuring beta-activity. Fluorescence activated cell sorting was used to quantify the SCLC cell surface expression of beta1-integrins. SETTING: Experiments were conducted in the Molecular Pharmacology Laboratory, Guthrie Research Institute. MEASUREMENTS AND RESULTS: Activation of mAChR with the agonist carbachol (10 microM, 1.5 h) significantly increases adhesion of the SCC-9 SCLC cell line to the ECM proteins laminin and collagen types I and IV. In contrast, mAChR activation does not alter the adhesion of SCC-9 cells to vitronectin, fibronectin, poly-L-lysine, or bovine serum albumin. Carbachol also does not alter the adhesion of NCI-H345 SCLC cells that lack functional mAChR. Preincubation of SCC-9 cells with the AIIB2 blocking antibody to beta1-integrin inhibits mAChR-induced adhesion to ECM proteins. Immunofluorescence analysis indicates that mAChR activation does not alter the surface expression of beta1-integrins by SCC-9 cells. Direct stimulation of protein kinase C (PKC) by treatment with phorbol 12-myristate 13-acetate (PMA) (10 nM, 1.5 h) increases the adhesion of both the SCC-9 and NCI-H345 cell lines to ECM proteins. These results indicate that direct activation of PKC or stimulation of M3 mAChR (which results in increased PKC activity) increases the binding activity of beta1-integrins, resulting in increased adhesion of SCLC cells to ECM proteins. CONCLUSIONS: The ability of mAChR to regulate SCLC proliferation and adhesion suggests that activation of these receptors may be used to alter SCLC tumorigenesis and metastasis. PMID- 9743177 TI - Four hours of continuous albuterol nebulization. AB - BACKGROUND AND OBJECTIVES: Continuous albuterol nebulization (CAN) is a therapeutic modality available to treat status asthmaticus. Currently, CAN may be administered using a large-volume nebulizer (LVN) or a small-volume nebulizer attached to an infusion pump or refilled as needed. Few data are available regarding the reproducibility of aerosol characteristics during CAN. In this study, we determined the aerodynamic profile, drug output (DO), DO in respirable range (RD), solution output (SO), and changes in reservoir's albuterol concentration (AR) hourly during 4 hours of CAN. DESIGN: A modified Puritan Bennett 1600 jet nebulizer was tested with a large reservoir (LR; 250 mL), medium reservoir (MR; 45 mL), and small reservoir with infusion pump (SRP; 18 mL). We used 100-, 40-, and 4-mL initial fill volumes (with 10-mL/h infusion for SRP) of 1 mg/mL albuterol solution for the LR, MR, and SRP, respectively. Particle size distribution and DO consistency were determined by impaction and spectrophotometric analysis (275 nm). We also determined albuterol mass output. The SO was determined by gravimetric technique. RESULTS: The PBsj produced a heterodisperse aerosol with a median mass aerodynamic diameter range of 1.8 to 2.2 microm. DO and RD paralleled SO. The LR had the highest SO, DO, and RD (8.03+/-2.36 vs 5.73+/-2.48 and 5.85+/-0.51 mg/h for MR and SRP, respectively). The AR showed no statistically significant changes. CONCLUSIONS: The PBsj demonstrated consistent and adequate aerosol production during 4 hours of CAN. These bench data support the widespread use of a LVN for CAN. PMID- 9743178 TI - The hemodynamic derangements in sepsis: implications for treatment strategies. AB - The incidence of the sepsis syndrome has increased dramatically in the last few decades. During this time, we have gained new insights into the pathophysiologic mechanisms leading to organ dysfunction in this syndrome. Yet, despite this increased knowledge and the use of novel therapeutic approaches, the mortality associated with the sepsis syndrome has remained between 30% and 40%. Appropriate antibiotic selection and hemodynamic support remain the cornerstone of treatment of patients with sepsis. Recent studies have failed to demonstrate a global oxygen debt in patients with sepsis. Furthermore, therapy aimed at increasing systemic oxygen delivery has failed to consistently improve patient outcome. The primary aim of the initial phase of resuscitation is to restore an adequate tissue perfusion pressure. Aggressive volume resuscitation is considered the best initial therapy for the cardiovascular instability of sepsis. Vasoactive agents are required in patients who remain hemodynamically unstable or have evidence of tissue hypoxia after adequate volume resuscitation. PMID- 9743179 TI - Utility of peak expiratory flow monitoring. PMID- 9743180 TI - Methodologic standards for diagnostic test research in pulmonary medicine. AB - STUDY OBJECTIVES: To determine conformance with methodologic standards in the evaluation of diagnostic tests. DATA SOURCES: MEDLINE database search (1992 to 1997) of nine prominent general medicine and six subspecialty journals for articles that report discriminative properties of diagnostic tests in pulmonary medicine. STUDY SELECTION: Articles were eligible if they reported discriminative properties of diagnostic tests in humans, diagnostic tests were intended for the detection of existing conditions, and the target disorder was relevant to pulmonary medicine. DATA EXTRACTION: Each study was critically reviewed independently by two observers. DATA SYNTHESIS: Of the 1,029 retrieved articles, 41 met study inclusion criteria. The median number of the 12 major standards for design fulfilled by study articles was 6 (range, 1 to 12, 25th to 75th percentile, 5.0 to 8.5) and only 2 articles fulfilled all 12 standards. Seven (17%) articles did not report any standard measures of diagnostic accuracy and 7 (17%) provided data only for sensitivity and specificity. Only 4 of 17 articles (24%) that compared different tests used standard statistical methods. CONCLUSION: These results indicate that greater methodologic rigor is needed for studies that evaluate diagnostic tests in pulmonary medicine. Existing deficiencies in methodology risk the introduction of invalid tests into clinical practice. PMID- 9743181 TI - Liberation from mechanical ventilation: a decade of progress. AB - Multiple complications associated with mechanical ventilation mandate that clinicians expeditiously define and reverse the pathophysiologic processes that precipitate respiratory failure and then, detect the earliest point that a patient can breathe without the ventilator. Over the past decade, numerous laboratory and clinical studies have been reported that may inform transformation of the "art of weaning" to the science of liberation. We review these studies and use them to formulate a systematic approach to assure early, safe, and successful liberation of patients from mechanical ventilation. PMID- 9743182 TI - Safety of medically supervised outpatient cardiac rehabilitation exercise therapy: a 16-year follow-up. PMID- 9743183 TI - Correction of single-breath helium lung volumes in patients with airflow obstruction. AB - STUDY OBJECTIVE: To determine whether alveolar volume (V(A)) measured during the single-breath diffusing capacity for carbon monoxide (DCO) can be used as a substitute measure for the multiple-breath total lung capacity (TLC) in subjects with and without airways obstruction. DESIGN: Retrospective review of pulmonary function test (PFT) results. SETTING: Pulmonary function laboratories at the Johns Hopkins Hospital (JHH) and the Johns Hopkins Asthma and Allergy Center (HAAC). PARTICIPANTS: Patients referred for spirometry, helium lung volumes, and Dco during a single visit between November 1993 and November 1996. RESULTS: JHAAC patients (n=2,477) were used to assess the relationship between V(A) and TLC. In patients with an FEV1/FVC > or = 0.70, there was good agreement between V(A) and TLC (V(A)/TLC=0.97 to 0.99). However, in patients with an FEV1/FVC <0.70, V(A) systematically underestimated TLC (V(A)/TLC=0.67 to 0.94). The degree of underestimation was related to the severity of airflow obstruction. To predict TLC using V(A) a regression equation was used to "correct" V(A) for the severity of obstruction. This equation was used to predict the multiple-breath TLC for JHH patients (n=2,892). Patients with an FEV1/FVC > or = 0.70 showed a high degree of correlation between V(A) and TLC (Pearson's correlation coefficient [r]=0.96 to 0.99; p<0.05). After adjusting for the severity of airflow obstruction, patients with an FEV1/FVC in the range of 0.40 to 0.70 also had a strong correlation between the corrected V(A) and the multiple-breath TLC (r=0.83 to 0.94; p<0.05). CONCLUSIONS: V(A) accurately predicts TLC in patients with mild or no airflow obstruction. For patients with moderate to severe obstruction, correcting V(A) for the severity of obstruction improves the accuracy of this relatively simple and rapid technique for measuring TLC. PMID- 9743184 TI - Recurrent ARDS in a 39-year-old woman with migraine headaches. PMID- 9743185 TI - Nonresolving pneumonia. PMID- 9743186 TI - Postpneumonectomy pulmonary edema: can it be predicted preoperatively? AB - Postpneumonectomy pulmonary edema (PPE) is a rarely reported form of acute lung injury which occurs in up to 4% of all pneumonectomies. The details of two well documented cases of PPE are described with special emphasis paid to the preoperative lung functions. Both cases illustrated a striking disparity between preserved spirometric lung function and advanced emphysema as detected by quantitative CT emphysema scores and single-breath diffusion of carbon monoxide measurements. Though retrospective in nature, these results suggest a restricted capillary volume plays a critical role in the development of PPE. PMID- 9743187 TI - Hypersensitivity pneumonitis due to humidifier disease: seek and ye shall find. AB - STUDY OBJECTIVES: This study reports a classic case of hypersensitivity pneumonitis (HP) with classic histologic changes in lung tissue and the research used to identify the causative antigens. DESIGN: A patient with clinical, radiographic, pulmonary function abnormalities and a lung biopsy consistent with HP had no identifiable antigen exposure. SETTING: Evaluation of the patient's activities provided no suggestion of antigen exposure. Her home was evaluated. It was found that her humidifier ran continually without being cleaned but water was added periodically. MEASUREMENTS: Serologic analysis demonstrated precipitating antibodies against her humidifier water and ten antigens in the hypersensitivity lung disease serologic panel. CONCLUSION: Removal of the humidifier, cleaning of the house, and a course of prednisone resulted in the return of the patient to a normal state. PMID- 9743188 TI - Paradoxical reactions in HIV and pulmonary TB. AB - We present a case of paradoxical clinical deterioration during antituberculosis therapy in an HIV-infected adult with pulmonary TB. The clinical course was characterized by marked cervical and mediastinal adenopathy accompanied by fever and weight loss during simultaneous treatment of TB and HIV disease. After extensive investigation for causes of therapeutic failure, the paradoxical reaction was attributed to partial immune reconstitution related to highly effective antiretroviral therapy. Due to the high prevalence of TB in HIV infected patients, it is important to recognize this phenomenon and understand that it is usually self-limited. PMID- 9743189 TI - Intralobar pulmonary sequestration with three aberrant arteries in a 75-year-old patient. AB - A rare case of intralobular pulmonary sequestration (ILS) with three aberrant arteries occurred in a 75-year-old woman. A contrast-enhanced chest CT scan demonstrated a paraaortic, partially enhanced mass shadow and two small liner enhancements in the upper portion of the mass. A definitive diagnosis could not be rendered with a CT scan alone, but the findings suggested bronchopulmonary sequestration with multiple aberrant arteries. Surgery confirmed three fine aberrant arteries arising from the thoracic aorta and entering the left lower lobe basal segment. Judging from the patient's age and multiple aberrant arteries, the sequestrated lung appeared as if it were acquired. However, all aberrant arteries were of the elastic type histologically. This finding suggested that ILS was not an acquired condition but a congenital malformation. PMID- 9743191 TI - Scoring organ dysfunction. PMID- 9743190 TI - Metastatic squamous cell carcinoma of the mediastinum with unknown primary tumor. AB - Although metastatic carcinoma from an unknown primary tumor is known to occur, the combination of squamous cell carcinoma histologic findings and a mediastinal location is quite unusual. The evaluation of a case of a patient with a posterior mediastinal mass, eventually shown to be metastatic squamous cell carcinoma of the mediastinum with unknown primary tumor, is described herein. Resection of the lymph node mass was performed and was followed by chemoradiation for presumed lung cancer. PMID- 9743192 TI - Theoretical basis for usefulness of dyspnea ratings for prescription of exercise in COPD patients. PMID- 9743193 TI - MIDCAB vs conventional surgery: is it worth the risk? PMID- 9743194 TI - Extrapulmonary manifestations of pneumonia. PMID- 9743195 TI - Improper use of MDIs: education is the key. PMID- 9743196 TI - Using the kappa coefficient as a measure of reliability or reproducibility. PMID- 9743197 TI - Share the research, not taxpayers' income. PMID- 9743198 TI - Tuberculin responsiveness in hemodialysis patients. PMID- 9743199 TI - Lack of HHV-8 DNA sequences in sarcoid tissues of French patients. PMID- 9743200 TI - Anticoagulating elderly patients in atrial fibrillation. PMID- 9743201 TI - Respiratory effects of the Dead Sea: a historical note. PMID- 9743202 TI - Levels of soluble L-selectin and E-selectin in heatstroke and heatstress. PMID- 9743203 TI - Effective implementation of practice guidelines. PMID- 9743204 TI - Polymorphism of the IL-1 gene complex in Epstein-Barr virus seronegative and seropositive adult blood donors. AB - Epstein-Barr virus (EBV) seronegativity is rare in adults. To examine whether genetic differences would explain this, we studied the genetic polymorphisms of the genes of the interleukin-1 (IL-1) complex in seronegative adults. These cytokines (i.e. IL-1alpha, IL-1beta and IL-1 receptor antagonist, IL-1RA) regulate, in several ways, the inflammatory reactions of the body. In each of these genes there are polymorphic sites and the various alleles differ in their frequency in several diseases of inflammatory nature. In 400 healthy blood donors (from 18 to 60 years of age) there were 20 (5%) seronegative persons. The frequency of allele 2 of the IL-1beta gene (base exchange polymorphism at position -511 from the transcriptional start site) was decreased in the seronegative patients (0.20 versus 0.42 in the seropositive patients, P < 0.05, chi2-test). Moreover, the frequency of allele 2 of the IL-1RA (polymorphism defined by variable numbers of 86-bp repeats in intron 2) was slightly, but not significantly, decreased in the seronegative patients. Alleles of these two loci are known to be associated, but in the seronegative patients this association was abnormal: 11 out of 20 (55%) were of the IL-1RA-2 negative/IL-1beta-2 negative type, while of the seropositive patients, 25% were of this type (P < 0.01, chi2 test). These data suggest that immunological differences, depending on cytokine gene polymorphisms, regulate the resistance to EBV infection. PMID- 9743205 TI - Comparison of the frequencies of arginines in heavy chain CDR3 of antibodies expressed in the primary B-cell repertoires of autoimmune-prone and normal mice. AB - Because the pathogenesis of anti-DNA Ab in SLE is correlated to Ab specificity for native DNA (dsDNA), understanding how such specificity is generated is important. The VH structures of most autoimmune anti-DNA antibodies include at least one arginine in VH-CDR3; moreover, antibody specificity for dsDNA can be correlated to the relative position of arginines in VH-CDR3. The coding sequences for most VH-CDR3 arginines among the anti-DNA MoAbs we have studied to date appeared to have been encoded by sequences generated during V-D-J recombination and would have been expressed in the primary B-cell repertoire. The frequency at which arginine codons are generated during V-D-J recombination therefore could potentially influence the frequency at which DNA-specific B cells are generated in the primary B-cell repertoire. The present study was undertaken to determine whether a higher percentage of B cells in the primary, preautoimmune repertoire of autoimmune-prone (NZB x NZW)F1 mice have immunoglobulin heavy chains with at least one VH-CDR3 arginine compared to B cells in the primary, preimmune repertoire of non-autoimmune-prone BALB/c mice. The present results indicate that mature B cells in preautoimmune (NZB x NZW)F1 mice, whether specific for DNA or not, are no more likely to have heavy chains with VH-CDR3 arginines than are B cells in BALB/c mice. The high frequency of recurrence of VH-CDR3 arginines among autoimmune anti-DNA in (NZB x NZW)F1 mice would appear to derive from the selective oligoclonal expansion of selected B cells that express such structures. PMID- 9743206 TI - Infection of B-cell-deficient mice by the parasite Schistosoma mansoni: demonstration of the participation of B cells in granuloma modulation. AB - The contribution of B lymphocytes to immunity towards the parasite Schistosoma mansoni has been investigated in a mouse strain rendered genetically B-cell deficient (the muMT mouse). These studies demonstrated that T cells primed in vivo in B-cell-deficient mice proliferate less efficiently in vitro in response to parasite antigenic extracts except at 10 weeks of infection. In addition, analysis of the cytokine profiles (IL-2, IL-4, IL-5 and IFN-gamma), investigated using RT-PCR, showed that spleens of muMT animals displayed a predominant Th1 like profile compared to control, B-cell-intact infected mice. This showed that B cells, either per se or through their secretions, are involved in the in vivo generation and/or maximal expansion of Th2-type T lymphocytes during the course of murine S. mansoni infection. Interestingly, the data showed that B-cell deficient mice display an increased hepatic fibrosis at 10 weeks postinfection (p.i.), whereas they behaved like infected controls, with regard to the other assessed parasitological parameters (e.g. worm burden estimation). This demonstrated that even if B lymphocytes are not essential for the development of the general immune response towards S. mansoni in the mouse, they may nevertheless be involved in the correct immunoregulation of the granulomatous reaction around the eggs. PMID- 9743207 TI - Catabolism of lipid-free recombinant apolipoprotein serum amyloid A by mouse macrophages in vitro results in removal of the amyloid fibril-forming amino terminus. AB - Serum amyloid A fibrils are formed when the normally rapid catabolism of the acute-phase reactant apolipoprotein serum amyloid A (apoSAA) is incomplete; thus amyloidosis may be viewed as a condition of dysregulated proteolysis. There is evidence that apoSAA is dissociated from plasma high-density lipoprotein (HDL) prior to fibril formation. The objective of this study was to investigate degradation of lipid-free apoSAA by tissue macrophages derived from amyloid susceptible CBA/J mice in vitro. Peritoneal macrophages derived from untreated (normal) mice converted apoSAA (12 kDa) to a single 4 kDa C-terminal peptide while splenic macrophages converted apoSAA to 10, 7 and 4 kDa C-terminal peptides and a 4 kDa peptide that lacked the C- and N-terminal regions. Similar patterns of proteolysis occurred when peritoneal and splenic macrophages from amyloidotic CBA/J mice were used. Conditioned medium prepared from peritoneal, but not splenic macrophages, degraded apoSAA. Specific sites of cleavage indicated activity of cathepsin G- and elastase-like neutral proteases. The data indicate that lipid-free apoSAA can be degraded by secreted or cell-associated neutral proteases that are generated by macrophages to yield peptides that lack fibrillogenic potential. PMID- 9743209 TI - Antioestrogens enhance tumour necrosis factor receptor 2 (TNF-R2) expression and TNF-R2-mediated proliferation in activated T cells. AB - In the present study we demonstrate that the non-steroidal antioestrogens toremifene and tamoxifen inhibit mitogen-induced proliferation and up-regulation of tumour necrosis factor (TNF) receptor family molecules on peripheral blood T cells. In activated T cells, however, toremifene and tamoxifen increase the surface expression of tumour necrosis factor receptor 2 (TNF-R2). This up regulation is functionally important as TNF-R2-mediated proliferation is significantly enhanced in antioestrogen-treated activated T cells. The regulation of TNF-R2 expression in activated T cells seems to involve the c-Jun amino terminal kinase (JNK) pathway, as activation of JNK with anisomycin down regulates TNF-R2. In activated T cells toremifene clearly inhibits phorbol 12 myristate 13-acetate (PMA)-induced JNK activity, suggesting that the JNK pathway may also be involved in the up-regulation of TNF-R2 expression by antioestrogens. Taken together, the enhancement of TNF-R2 expression and TNF-R2-mediated proliferation in activated T cells represents a novel feature for the effects of antioestrogens. The inhibitory effects of toremifene on the JNK pathway demonstrates that antioestrogens can influence not only cell growth, but also a variety of other cellular responses by inhibiting protein kinase C (PKC). PMID- 9743208 TI - Increased PGE2 production mediates the in vitro inhibitory effect of the human immunodeficiency virus P24 immunosuppressive heptapeptide Ch7. AB - Previous work from our laboratory demonstrated that a synthetic heptapeptide (Ch7), corresponding to a conserved sequence of human immunodeficiency virus (HIV) core protein p24 (amino acids 232-238), was able to specifically abrogate antigen-induced responses in cultures of normal human peripheral blood lymphocytes (PBL). Addition of recombinant human interferon-gamma (IFN-gamma) to Ch7-suppressed cultures restored the capacity to mount an antigen-specific antibody response, suggesting that a cytokine imbalance may be at the basis of the Ch7 immunosuppressive activity. In the present paper we show that the Ch7 dependent in vitro immunosuppression was accompanied by a significant up regulation of prostaglandin E2 (PGE2) production and induction of interleukin-10 (IL-10)-secreting cells. In the presence of the PGE2 inhibitor indomethacin, IL 10 up-regulation was prevented and the induction of a specific antibody response was partially restored. PGE2 is indeed an important regulator of immune responses with the ability to differentially affect cytokine production. Thus, our results demonstrate that the Ch7 immunosuppressive epitope may primarily act by up regulating PGE2 production and, through this mediator, by causing a cytokine dysregulation, finally responsible for immune response suppression. PMID- 9743210 TI - Characterization of soluble terminal complement complex assembled in C8beta deficient plasma and serum. AB - Sera genetically deficient in either the alpha-gamma or the beta-subunit of complement component C8 virtually lack haemolytic activity. We have studied the formation and the structural organization of the soluble terminal complement complex (TCC) assembled in these sera following activation with cobra venom factor (CVF). The TCC concentration in the activated C8alpha-gamma and C8beta deficient samples was 0.2% and 4%, respectively, when compared with zymosan activated normal serum. TCC was purified from the activated C8beta-deficient samples by affinity chromatography and analysed by immunoblotting and enzyme immunoassay. No C8beta was detected in one TCC preparation, while 7% of the normal level was present in the other. The level of the other terminal components, including that of C8alpha-gamma, was normal. The ability of C8alpha gamma to promote the assembly of TCC in the presence of a limited amount of C8beta or in the apparent absence of this subunit was confirmed using purified components, by mixing C5b6 and either of the purified C8 subunits together with C7 and C9. These data show that soluble TCC can be formed in C8beta-deficient sera that contain little or no C8beta. PMID- 9743211 TI - Prostaglandin-independent stimulation of interleukin-6 production by fibrinogen degradation product D in perfused murine liver. AB - Bacterial endotoxin (LPS) and fibrinogen degradation product D (FDP-D) are both potent stimulators of interleukin-6 (IL-6) production in liver, however, there are differences in their metabolic effects. The aim of the present study was to compare the role of prostaglandins in the enhancement of IL-6 production by LPS or FDP-D in perfused mouse livers. Indomethacin inhibited the effect of LPS significantly but was ineffective in the case of FDP-D. Accordingly, production of prostaglandins D2 and E2 was not elevated following the addition of FDP-D, while their formation was increased several fold by LPS. At the same time interleukin-1 (IL-1) production in perfused liver rose markedly upon the addition of FDP-D. It is suggested that prostaglandins are not involved in the effects of FDP-D on the liver. The stimulatory effect of FDP-P on IL-6 production might be the consequence of elevated IL-1 levels. PMID- 9743212 TI - Acute, lethal graft-versus-host disease in an F1-hybrid model using grafts from parental-strain, T-cell receptor-delta gene knockout donors. AB - Gammadelta T cells have been implicated in the pathogenesis of acute graft-versus host disease (GVHD). We therefore performed experiments to determine whether mortality from GVHD is reduced in C57BL/6 x DBA/2 F1-hybrid (BDF1-hybrid) mice when parental strain, T-cell receptor-delta (TCRdelta) knockout (KO) donors are used. We compared mortality, weight loss, interferon-gamma (IFN-gamma) production and cytotoxic activity in recipients of either wild-type or TCRdelta KO grafts. In both groups there was significant weight loss and an identical level of mortality. Elevated IFN-gamma levels were present in both groups, but recipients of TCRdelta KO grafts produced twice as much as recipients of wild-type grafts. Elevated natural killer (NK) and NK-like activity was also seen in both. These results demonstrate that TCRdelta KO grafts can induce GVHD as severe as that seen in recipients of wild-type grafts, a finding that is at odds with studies demonstrating reduced mortality when gammadelta T cells are purged from donor mice. We suggest that the inconsistency may lie in the higher levels of IFN-gamma seen with TCRdelta KO grafts and that the protection afforded by the absence of gammadelta T cells in the graft is overwhelmed by the higher levels of IFN-gamma. PMID- 9743213 TI - Increased antibody production against gut-colonizing Escherichia coli in the presence of the anaerobic bacterium Peptostreptococcus. AB - Germ-free rats were colonized with E. coli alone, or with E. coli plus Lactobacillus acidophilus and a strain of the obligate anaerobic gram-positive species, Peptostreptococcus. The presence of Peptostreptococcus reduced translocation of E. coli, but increased the serum antibody response to E. coli antigen. Whereas the immunoglobulin G (IgG) anti-E. coli antibodies largely represented cross-reactive antibodies, those of the immunoglobulin M (IgM) isotype represented true anti-E. coli antibodies because they could not be absorbed by L. acidophilus or Peptostreptococcus but could with E. coli. We suggest that peptostreptococci prime the gut immune system to other bacterial antigens and that this could be a mechanism behind the reduced translocation of facultative anaerobes in the presence of obligate anaerobes. PMID- 9743214 TI - Neonatal tolerance revisited by mathematical modelling. AB - The classical view of a neonatal tolerance window has recently been challenged by several studies showing that neonates can evoke normal immune responses. For example, neonatal immunity against the male antigen H-Y can be induced in female recipients by inoculation of male donor spleen cells enriched for professional antigen-presenting cells (APCs). In the same set-up, adult female recipients become tolerant by giving large doses of spleen cells. Using a probabilistic model, we here show how the number of T cells and endogenous APCs in the recipient, and the dose and quality of donor APCs can explain all observed phenomena. We thus reconcile the classical neonatal tolerance window with the recent data on neonatal immunity and adult tolerance. PMID- 9743215 TI - Synovial fluid cytokines in patients with rheumatoid arthritis or other arthritic lesions. AB - The synovial fluid (SF) of rheumatoid arthritis (RA) patients contains a mixture of inflammatory mediators. In order to determine whether certain cytokine patterns locally in the joint are specifically related to the chronic inflammation in RA, the concentrations of interleukin (IL)-1alpha, IL-1beta, IL 6, IL-10, transforming growth factor-beta (TGF-beta), tumour necrosis factor alpha (TNF-alpha) and IgG2b-inducing factor (IgG2bIF) were measured in SF from 22 patients with RA and 22 patients with other types of arthritic lesions. High levels of IL-10, latent and active TGF-beta and the presence of IgG2bIF are significantly correlated with RA when corrected for age. As these factors have the capacity to promote antibody production, they might contribute to the maintenance of local antibody production in RA synovial tissues. All RA-SF samples contained detectable levels of IL-10 and all except one contained IL 1beta, while concentrations in several non-RA-SF samples were below detection limits. IL-6 and TGF-beta were present in all SF samples from both RA and non-RA patients. The presence of IgG2bIF was strongly correlated with high levels of IL 10 and IL-1beta in SF. However, no distinct cytokine profile specific for the chronic inflammation characteristic of RA was found. PMID- 9743216 TI - Autoantibody patterns in synovial fluids from patients with rheumatoid arthritis or other arthritic lesions. AB - Patients with rheumatoid arthritis (RA) produce a variety of autoantibodies, not only demonstrable in the circulation, but also locally in the inflamed joint. We investigated the local production of several autoantibodies in the synovial fluid (SF) of 24 patients with RA and of 26 patients with other arthritic lesions. RA patients had higher titres of immunoglobulin M (IgM) and immunoglobulin G (IgG) rheumatoid factors (RFs) and of collagen type II antibodies in SF, whereas there were no demonstrable differences between groups with regard to antibodies against double-stranded (ds) DNA, C1q or the hapten 2,4,6-trinitrobenzene sulfonic acid (TNP). No differences were observed for total synovial levels of IgM or IgG. There was no autoantibody pattern that was typical of RA patients, except for the local presence of RF, primarily in seropositive RA patients. Our findings therefore support the notion that RF and collagen type II antibodies are induced by immunogenic material present in the local inflamed environment. In the accompanying paper we studied various synovial fluid cytokines in the same patient groups. Here we correlated the level of these cytokines with autoantibody titres in SF, but no specific cytokine associated with the production of RF was found. Hence, we conclude that several different inflammatory mediators might contribute to the chronic inflammation and autoantibody production in the joint of RA patients. An inverse correlation was established between concentrations of tumour necrosis factor-alpha (TNF-alpha) and levels of total IgG. PMID- 9743217 TI - T-cell clonal change after allo-kidney transplantation in humans. AB - Whether T cells circulating peripherally express changes at a clonal level after renal transplantation is uncertain. To clarify this issue, we analyzed T-cell clonality of peripheral blood lymphocytes (PBLs) in 12 renal transplant recipients by a novel polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) method that can discriminate T-cell clones with different T-cell receptor (TCR) Vbeta motifs. The PCR-SSCP study showed that after transplantation, only a few distinct T-cell clonotypes accumulated in the absence of clinical episodes, irrespective of the compatibility of HLA antigens. In contrast, various T-cell clones appeared in cases of acute rejection (AR) and infection. These subsided immediately after the AR was resolved; however, they remained long after the resolution of the infection. In a case of AR followed by an infectious episode, distinct T-cell clones appeared concomitantly with each episode. Several of them disappeared or remained thereafter. In one case, significant numbers of accumulating bands were observed by in-vitro stimulation by mixed lymphocyte reaction (MLR); several were identical to those found in vivo. However, some of those that did not appear in vitro were apparent in vivo. In conclusion, the appearance of T-cell clonotypes at a peripheral level indicates the existence of immunologically activated T-cell clones, which were significantly affected by immunosuppressive therapy. It was also determined that the T-cell immune system is much more complicated in vivo than in vitro. PMID- 9743218 TI - Correlation of Fc gamma receptor expression of monocytes with clearance function by macrophages in systemic lupus erythematosus. AB - The expression of Fc gamma receptor II (Fc gammaRII) and Fc gammaRIII on monocytes in peripheral blood and the clearance of immunoglobulin (Ig)G sensitized erythrocytes (EA) by tissue macrophages were investigated in parallel in patients with systemic lupus erythematosus (SLE). The relationship between receptor expression and the rate of clearance of EA (half-time) was analyzed. The detected decrease in mean fluorescence intensity of both FcR gammaII and Fc gammaRIII of patients' monocytes stained with specific monoclonal antibodies (IV.3 and 3G8) was inversely correlated with the prolonged clearance half-time of 51Cr-labelled and anti-D IgG-sensitized autologous erythrocytes in these patients. A correlation was found between the impaired clearance function and the severity of the disease manifestation expressed by either clinical activity or renal involvement in our SLE patients. From these results it can be concluded that the in-vitro determination of monocyte Fc gammaRII and Fc gammaRIII expression may predict the in-vivo macrophage function via the same Fc receptors. PMID- 9743219 TI - High-dose intravenous immunoglobulin treatment activates complement in vivo. AB - Several complement modulating effects of high-dose intravenous immunoglobulins (IVIG) have been proposed from in vitro studies and experimental animal models. However, the in vivo effects of IVIG on plasma complement in humans are yet not known. We have investigated the in vivo effects of IVIG on complement in seven women with unexplained recurrent spontaneous abortion who were without evidence of autoimmune disease. Samples were obtained before and after the very first infusion of IVIG. There was a marked increase in immunoglobulin G (IgG) from (median and range) 12.4 (9.4-15.9) to 26.8 (22.4-30.0) g/l but no change in immunoglobulin A (IgA) or immunoglobulin M (IgM). A significantly increased complement activation was demonstrated using neoepitope-specific enzyme immunoassays to the activation products C3bc (median increased from 9.8 to 31.2 AU/ml), Bb (0.66-1.66 g/ml), C5a (10.5-12.7 ng/ml), and TCC (0.81-2.19 AU/ml) (P = 0.015 for all). There were no changes in antigenic concentrations of individual complement components or regulators (C1q, C4, C3, C1-inhibitor, C4b-binding protein) and no decrease in complement haemolytic activity (classical and alternative CH50), which were all within the normal range. The classical pathway activation products C1rs/C1-inhibitor complexes, C4bc and C4d were elevated in all patients before IVIG treatment and did not change significantly during treatment. In conclusion, IVIG induced a significant activation of complement in vivo. PMID- 9743220 TI - Variation in gut-homing CD27-negative lymphocytes in inflammatory colon disease. AB - Prolonged antigenic stimulation results in lymphocyte shedding of CD27, a member of the tumour necrosis factor receptor (TNFR) family, and transformation to a stable phenotype capable of synthesizing interleukin-4 (IL-4). Co-expression of alpha4beta7 identifies those cells with gut-homing potential. We have investigated these cell populations in patients with inflammatory colonic disease. Circulating and lamina propria mononuclear cells were isolated from patients with Crohn's disease (CD), ulcerative colitis (UC), non-inflammatory bowel disease (non-IBD) colonic inflammation and healthy controls. Double and triple colour flow cytometry for CD3, CD4, CD27, alpha4beta7 and intracellular cytokines was performed. Circulating CD4+ CD27- populations were increased in patients with CD (8.8 +/- 0.8%, P < 0.001), UC (12.2 +/- 1.9%, P < 0.001) and non IBD colitis (10.5 +/- 1.3%, P < 0.01) as compared with controls (6.1 +/- 0.5%). CD4+ CD27- alpha4beta7+ cells were increased in CD (P < 0.01). Lamina propria CD4+ CD27- populations were depressed significantly in CD (P < 0.05), UC (P < 0.02) and non-IBD colitis (P < 0.03). Mucosal CD4+ CD27- cells synthesized IL-4 in preference to interferon-gamma. Thus, colonic inflammation is associated with alterations in gut-tropic circulating and mucosal populations of differentiated memory T cells with the phenotype of predominantly IL-4-synthesizing cells. PMID- 9743221 TI - Immunophenotypic characteristics of monocytes in elderly subjects. AB - Since ageing is accompanied by various alterations of the immune system, the aim of the present study was to investigate the effect of age on the expression of surface markers in peripheral blood monocytes. We studied 28 healthy young subjects and 28 elderly subjects fulfilling the criteria of the SENIEUR protocol. Peripheral blood mononuclear cells were isolated and the expression of various surface markers was analysed by double colour flow cytometry. The mean fluorescence intensity of the intercellular adhesion molecule- (CD54), CD29, CD45RO and CD32 was increased significantly in CD14+ cells of elderly people when compared with the young subjects. No significant differences were found in the expression of CD11a, CD11b, CD15, CD26, CD27, CD33, CD45RA, CD45RB, CD49d, HLA-DR and CD65. In summary, we demonstrated significant increase in four monocyte surface markers in subjects of old age; this finding may be related to certain immune phenomena observed in ageing subjects such as auto-immunity. PMID- 9743222 TI - von Willebrand factor: a marker of endothelial damage? PMID- 9743223 TI - Fibronectin: structure, assembly, and cardiovascular implications. PMID- 9743224 TI - Plasma levels of activated protein C in healthy subjects and patients with previous venous thromboembolism: relationships with plasma homocysteine levels. AB - The proteolytic enzyme activated protein C (APC) is a normal plasma component, indicating that protein C (PC) is continuously activated in vivo. High concentrations of homocysteine (Hcy) inhibit the activation of PC in vitro; this effect may account for the high risk for thrombosis in patients with hyperhomocysteinemia (HyperHcy). We measured the plasma levels of APC in 128 patients with previous venous thromboembolism (VTE) and in 98 age- and sex matched healthy controls and correlated them with the plasma levels of total Hcy (tHcy) measured before and after an oral methionine loading (PML). Forty-eight patients had HyperHcy and 80 had normal levels of tHcy. No subject was known to have any of the congenital or acquired thrombophilic states at the time of the study. Because the plasma levels of APC and PC were correlated in healthy controls, the APC/PC ratios were also analyzed. Plasma APC levels and APC/PC ratios were significantly higher in VTE patients than in controls (P=0.03 and 0.0004, respectively). Most of the increase in APC levels and APC/PC ratios were attributable to patients with HyperHcy. Patients with normal tHcy had intermediate values, which did not differ significantly from those of healthy controls. There was no correlation between the plasma levels of tHcy or its PML increments and APC or APC/PC ratios in controls. The fasting plasma levels of APC and APC/PC ratios of 10 controls did not increase 4 hours PML, despite a 2-fold increase in tHcy. This study indicates that APC plasma levels are sensitive markers of activation of the hemostatic system in vivo and that Hcy does not interfere with the activation of PC in vivo. PMID- 9743225 TI - Atherogenic properties of enzymatically degraded LDL: selective induction of MCP 1 and cytotoxic effects on human macrophages. AB - The mechanisms underlying the selective accumulation of macrophages in early atherosclerotic lesions are poorly understood but are likely to be related to specific properties of altered low density lipoprotein (LDL) deposited in the subendothelium. Enzymatic, nonoxidative degradation of LDL converts the lipoprotein to a potentially atherogenic moiety, enzymatically altered LDL (E LDL), which activates complement and is rapidly taken up by human macrophages via a scavenger receptor-dependent pathway. Immunohistological evidence indicates that E-LDL is present in an extracellular location in the early lesion. We report that E-LDL causes massive release of monocyte chemotactic protein 1 (MCP-1) from macrophages and that expression of interleukin 8 or RANTES remains unchanged. Release of MCP-1 was preceded by a rapid expression of MCP-1 mRNA, which was detectable after 15 minutes, reached maximum levels after 1 hour, and remained detectable for 12 hours after exposure to concentrations as low as 10 microg/mL E LDL. MCP-1 mRNA induction and protein release by E-LDL exceeded that evoked by oxidized LDL. Release of MCP-1 was dependent on de novo protein synthesis and on the activity of tyrosine kinases. At higher concentrations, E-LDL, but not oxidized LDL, exerted toxic effects on macrophages that in part appeared to be due to apoptosis. The results show that E-LDL possesses major properties of an atherogenic lipoprotein. PMID- 9743226 TI - C-reactive protein frequently colocalizes with the terminal complement complex in the intima of early atherosclerotic lesions of human coronary arteries. AB - There is increasing evidence that complement activation may play a role in atherogenesis. Complement proteins have been demonstrated to be present in early atherosclerotic lesions of animals and humans, and cholesterol-induced atherosclerotic lesion formation is reduced in complement-deficient animals. Potential complement activators in atherosclerotic lesions are now a subject matter of debate. C-reactive protein (CRP) is an acute-phase protein that is involved in inflammatory processes in numerous ways. It binds to lipoproteins and activates the complement system via the classic pathway. In this study we have investigated early atherosclerotic lesions of human coronary arteries by means of immunohistochemical staining. We demonstrate here that CRP deposits in the arterial wall in early atherosclerotic lesions with 2 predominant manifestations. First, there is a diffuse rather than a focal deposition in the deep fibroelastic layer and in the fibromuscular layer of the intima adjacent to the media. In this location, CRP frequently colocalizes with the terminal complement complex. Second, the majority of foam cells below the endothelium show positive staining for CRP. In this location, no colocalization with the terminal complement proteins can be observed. Our data suggest that CRP may promote atherosclerotic lesion formation by activating the complement system and being involved in foam cell formation. PMID- 9743227 TI - Apolipoprotein(a) enhances platelet responses to the thrombin receptor-activating peptide SFLLRN. AB - Elevated levels of lipoprotein(a) [Lp(a)] are correlated with an increased risk of atherosclerotic disease. We examined the effect of recombinant apolipoprotein(a) [r-apo(a)] and Lp(a) on responses of washed human platelets, prelabeled in the dense granules with [14C]serotonin and suspended in Tyrode's solution, to ADP and the thrombin receptor-activating peptide SFLLRN. No effect of the 17 kringle (K), 12K, or 6K r-apo(a) derivatives (at concentrations of 0.35 and 0.7 micromol/L) or Lp(a) (up to 0.1 micromol/L) on primary ADP-induced platelet aggregation was observed. In contrast, weak platelet responses stimulated by 7.5 micromol/L SFLLRN were significantly enhanced by the r-apo(a) derivatives; eg, 0.7 micromol/L 17K r-apo(a) increased aggregation from 15+/-4% to 58+/-6%, release of [14C]serotonin from 9+/-3% to 36+/-6%, and formation of thromboxane A2, measured as its stable metabolite thromboxane B2, from 7+/-1 to 29+/-5 ng/10(9) platelets (n=3; P<0.04 to 0.015). Significant enhancement of aggregation and release of granule contents was observed at a concentration of 17K r-apo(a) as low as 0.175 micromol/L. Purified Lp(a) (0.25 to 0.1 micromol/L) also enhanced SFLLRN-induced aggregation and release in a dose-dependent manner. Although plasminogen (0.7 and 1.5 micromol/L) and low density lipoprotein (0.025 to 0.1 micromol/L) both exhibited potentiating effects on SFLLRN-mediated platelet aggregation, the magnitude of the responses was less than that observed with either the r-apo(a) derivatives or Lp(a). The enhanced responses of platelets via the protease-activated receptor- thrombin receptor in the presence of Lp(a) may contribute to the increased risk of thromboembolic complications of atherosclerosis associated with this lipoprotein. PMID- 9743228 TI - Warfarin causes rapid calcification of the elastic lamellae in rat arteries and heart valves. AB - High doses of warfarin cause focal calcification of the elastic lamellae in the media of major arteries and in aortic heart valves in the rat. Aortic calcification was first seen after 2 weeks of warfarin treatment and progressively increased in density at 3, 4, and 5 weeks of treatment. By 5 weeks, the highly focal calcification of major arteries could be seen on radiographs and by visual inspection of the artery. The calcification of arteries induced by warfarin is similar to that seen in the matrix Gla protein (MGP)-deficient mouse, which suggests that warfarin induces artery calcification by inhibiting gamma carboxylation of MGP and thereby inactivating the putative calcification inhibitory activity of the protein. Warfarin treatment markedly increased the levels of MGP mRNA and protein in calcifying arteries and decreased the level of MGP in serum. Warfarin treatment did not affect bone growth, overall weight gain, or serum calcium and phosphorus levels, and, because of the concurrent administration of vitamin K, prothrombin times and hematocrits were normal. The results indicate that the improved warfarin plus vitamin K treatment protocol developed in this study should provide a useful model to investigate the role of MGP in preventing calcification of arteries and heart valves. PMID- 9743229 TI - Chronic blockade of NO synthase activity induces a proinflammatory phenotype in the arterial wall: prevention by angiotensin II antagonism. AB - Chronic blockade of NO production induces hypertension and early occlusive and fibrotic end-stage organ damage owing to vascular lesions in the brain, kidney, and heart. In this study, we evaluated the inflammatory phenotypic changes induced in the arterial wall by chronic N(G)-nitro-L-arginine methyl ester (L NAME) administration and the effect of an angiotensin II receptor (AT1) antagonist, irbesartan, on these changes. For this purpose, 2 groups of rats received L-NAME in the drinking water (50 mg x kg(-1) x d(-1)) for 2 months. One group received no other treatment and the other was treated with irbesartan (10 mg x kg(-1) x d(-1)). A third group (controls) received neither L-NAME nor irbesartan. After 8 weeks, plasma, aortas, and left ventricles were sampled from all 3 groups. Expression of inducible NO synthase (iNOS) was evaluated at both the mRNA (quantitative reverse transcription-polymerase chain reaction) and the protein (Western blot and immunohistochemistry) level in the aorta. Expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) was evaluated by reverse transcription-polymerase chain reaction, Western immunoblotting, and immunohistochemistry; inflammatory cell infiltration by immunohistochemistry; and fibrosis by Sirius red staining. Chronic L-NAME administration induced the expression of iNOS in the aorta, which was localized in smooth muscle cells as shown by immunohistochemistry and NADPH diaphorase activity. ICAM-1 and VCAM-1 expression was also increased in aortas of L-NAME treated rats. These phenotypic changes of the vascular wall were associated with inflammatory cell infiltration and fibrosis in the heart. All of these pathological phenomena were prevented by the angiotensin II antagonist irbesartan. The proinflammatory phenotypic changes of the vascular wall induced by blockade of NOS activity could be involved in the interaction between endothelial dysfunction and the development of arteriosclerosis. PMID- 9743230 TI - Human apolipoproteins A-I and A-II in cell cholesterol efflux: studies with transgenic mice. AB - The first step in reverse cholesterol transport is the movement of cholesterol out of cells onto lipoprotein acceptors in the interstitial fluid. The contribution of specific lipoprotein components to this process remains to be established. In this study, the role of human apolipoproteins (apo) A-I and A-II in the efflux of cellular cholesterol was investigated in transgenic mouse models in which the expression of murine apoA-I was abolished due to gene targeting (A IKO). Serum from A-IKO mice and from mice expressing human apoA-I and/or human apoA-II was incubated with [3H]cholesterol-labeled Fu5AH rat hepatoma cells for 4 hours at 37 degrees C. The cholesterol efflux to the serum of A-IKO mice was markedly lower than that to the serum of mice transgenic for human apoA-I (5.0 +/ 1.5% versus 25.0 +/- 4.0%). Expression of human apoA-II alone did not modify the cholesterol efflux capacity of A-IKO mouse serum. Cholesterol efflux to serum of mice expressing human apoA-II together with human apoA-I was significantly lower than that to human apoA-I mouse serum (20.0 +/- 2.3% versus 25.0 +/- 4.0%). Regression analysis of cholesterol efflux versus the lipid/apolipoprotein concentrations of mouse serum suggested that 3 independent factors contribute to determine the cholesterol efflux potential of serum: the apolipoprotein composition of HDL, the serum concentration of HDL phospholipids, and the presence of a small fraction of particles containing apoA-I. PMID- 9743231 TI - Enhanced upregulation of the Fc gamma receptor IIIa (CD16a) during in vitro differentiation of ApoE4/4 monocytes. AB - We recently reported a positive correlation of the pool size of lipopolysaccharide receptor (CD14)dim and Fc gamma receptor IIIa (CD16a)+ monocytes in peripheral blood to the apolipoprotein E4 (apoE4) phenotype and a negative correlation to high density lipoprotein (HDL) cholesterol levels (Arterioscler Thromb Vasc Biol. 1996;16:1437-1447). In this study, the in vitro differentiation of mononuclear phagocytes derived from healthy blood donors homozygous for the E3/3 or the E4/4 phenotype was analyzed during 7 days of culture in serum-free medium supplemented with macrophage colony-stimulating factor (M-CSF). The CD16a expression, which indicates Fc receptor-dependent phagocytic activity, increased to a significantly higher level in apoE4/4 monocytes than in apoE3/3 cells. The costimulatory molecule CD40, which indicates antigen-presenting capacity, was upregulated more strongly in apoE3/3 monocytes compared with E4/4 cells, but the difference did not reach a significant level. The expression of differentiation-associated surface proteins (CD14, CD33, CD45) and adhesion molecules (CD11a, CD11b, CD11c, CD49d) was not significantly different between apoE3/3 and apoE4/4 monocytes. However, a significantly decreased intracellular apoE concentration and a reduced amount of secreted apoE were found in apoE4/4 monocytes during in vitro differentiation. No differences were found in the surface expression of the low density lipoprotein receptor related protein (CD91) and the uptake of fluorescence labeled low density lipoprotein between apoE3/3 and apoE4/4 monocytes. These data indicate that the apoE4/4 phenotype significantly influences the M-CSF-dependent differentiation of monocytes toward a more CD16a-positive phagocytic phenotype. PMID- 9743232 TI - Increased angiotensin II type 1 receptor expression in hypercholesterolemic atherosclerosis in rabbits. AB - Angiotensin II (Ang II) promotes vascular smooth muscle growth and may be involved in the initiation and progression of atherosclerosis. To examine whether Ang II receptor expression in vascular tissues is altered in atherosclerosis, male New Zealand White rabbits were fed a high-cholesterol diet (1% cholesterol + 4% coconut oil mixed with regular chow; hypercholesterolemic group, n=12) or regular chow (control group, n=8) for 10 weeks. At the end of this period, the serum cholesterol level in the rabbits fed the high-cholesterol diet was higher than that in the control group (3616 +/- 144 versus 30 +/- 1 mg/dL, P<0.001). There was no atherosclerosis in the aortas of the control group, whereas 51 +/- 6% of the aorta was covered with atherosclerosis in the hypercholesterolemic group. Total Ang II receptor expression in the atherosclerotic aortic tissues was increased 5-fold in the hypercholesterolemic rabbits (292 +/- 28 versus 51 +/- 32 fmol/mg tissue, mean +/- SE, P<0.001), and the increased Ang II receptor expression was entirely due to enhanced Ang II type 1 (AT1) receptor expression (289 +/- 38 versus 38 +/- 18 fmol/mg, P<0.001), as Ang II type 2 receptor expression was unaltered (7 +/- 5 versus 3 +/- 2 fmol/mg, P=NS). AT1 receptors were localized primarily in the media and to some extent in the intima of the atherosclerotic aorta, as determined by immunohistochemistry with specific monoclonal and polyclonal AT1 receptor antibodies. Increased synthesis of AT1 receptor mRNA in atherosclerotic tissues was confirmed by reverse transcription polymerase chain reaction. To evaluate the functional significance of increased AT1 receptor expression, the constrictor response of aortic rings to Ang II was examined and found to be markedly enhanced in atherosclerotic aortic rings (P<0.01 versus control aortic rings). The endothelium-dependent relaxation of aortic rings from hypercholesterolemic rabbits was markedly attenuated (P<0.001). This study shows that hypercholesterolemia in rabbits results in atherosclerosis, loss of endothelium-dependent relaxation, and increased Ang II receptor (entirely AT1 receptor) expression in aortic tissues, which may result in altered vasoreactivity. PMID- 9743233 TI - Transcriptional activation of scavenger receptor expression in human smooth muscle cells requires AP-1/c-Jun and C/EBPbeta: both AP-1 binding and JNK activation are induced by phorbol esters and oxidative stress. AB - Reactive oxygen species generated by treatment of smooth muscle cells (SMCs) with either phorbol 12-myristate 13-acetate or with the combination of H2O2 and vanadate strongly induce expression of the class A scavenger receptor (SR-A) gene. In the current studies, cis-acting elements in the proximal 245 bp of the SR-A promoter were shown to direct luciferase reporter expression in response to oxidative stress in both SMCs and macrophages. A composite activating protein-1 (AP-1)/ets binding element located between -67 and -50 bp relative to the transcriptional start site is critical for macrophage SR-A activity. Mutation of either the AP-1 or the ets component of this site also prevented promoter activity in SMCs. Mutation of a second site located between -44 and -21 bp, which we have identified as a CCAAT/enhancer binding protein (C/EBP) element, reduced the inducible activity of the promoter in SMCs by 50%, suggesting that combinatorial interactions between these sites are necessary for optimal gene induction. Interactions between SMC nuclear extracts and the SR-A promoter were analyzed by electrophoretic mobility shift assay. c-Jun/AP-1 binding activity, specific for the -67- to -50-bp site, was induced in SMCs by the same conditions that increased SR-A expression. Moreover, phorbol 12-myristate 13-acetate, H2O2, or the combination of H2O2 and sodium orthovanadate (vanadate) activated c-Jun activating kinase. The binding activity within SMC extracts specific for the C/EBP site was shown to be C/EBPbeta in SMCs. Taken together, these findings demonstrate that reactive oxygen species regulate the interactions between c Jun/AP-1 and C/EBPbeta in the SR-A promoter. Furthermore, induction of oxidative stress in THP-1 cells, with a combination of 10 micromol/L vanadate and 100 micromol/L H2O2, induced macrophage differentiation, adhesion, and SR activity. These data suggest that vascular oxidative stress may contribute to the induction of SR-A expression and thereby promote the uptake of oxidatively modified low density lipoprotein by both macrophage and SMCs to produce foam cells in atherosclerotic lesions. PMID- 9743234 TI - Factors influencing the ability of HDL to inhibit expression of vascular cell adhesion molecule-1 in endothelial cells. AB - We have previously reported that high density lipoproteins (HDLs) inhibit the cytokine-induced expression of adhesion molecules in endothelial cells. Here we investigate whether different preparations of HDLs vary in their ability to inhibit the expression of vascular cell adhesion molecule-1 (VCAM-1) in human umbilical vein endothelial cells (HUVECs) activated by tumor necrosis factor alpha (TNF-alpha). HDLs collected from a number of different human subjects all inhibited VCAM-1 expression in a concentration-dependent manner, although the extent of inhibition varied widely between subjects. The inhibitory activities of the HDL2 and HDL3 subfractions isolated from individual subjects also differed. Whether equated for concentrations of apolipoprotein (apo) A-I or cholesterol, the inhibitory activity of HDL3 was superior to that of HDL2. This difference remained apparent even when the HDL subfractions were present only during preincubations with the HUVECs and were removed before activation by TNF-alpha. To determine whether the inhibitory effect of HDL3 was influenced by apolipoprotein composition, preparations of HDL3 were modified by replacing all of their apo A-I with apo A-II. This change in apolipoprotein composition had no effect on the ability of the HDL3 to inhibit endothelial VCAM-1 expression. Thus, it has been shown that different preparations of HDLs differ markedly in their abilities to inhibit VCAM-1 expression in cytokine-activated HUVECs. The mechanism underlying the differences remains to be determined. PMID- 9743235 TI - Inhibition of NO synthesis induces inflammatory changes and monocyte chemoattractant protein-1 expression in rat hearts and vessels. AB - We recently showed that chronic inhibition of NO synthesis by N(omega)-nitro-L arginine methyl ester (L-NAME) causes coronary vascular remodeling (ie, vascular fibrosis and medial thickening) in rats. To test the hypothesis that the inhibition of NO synthesis induces inflammatory changes in the heart, we characterized the inflammatory lesions that occurred during L-NAME administration and determined whether inflammation involved the induction of monocyte chemoattractant protein-1 (MCP-1) in vivo. During the first week of L-NAME administration to Wistar-Kyoto rats, we observed a marked infiltration of mononuclear leukocytes (ED1-positive macrophages) and fibroblast-like cells (alpha-smooth muscle actin-positive myofibroblasts) into the coronary vessels and myocardial interstitial areas. These inflammatory changes were associated with the expression of proliferating cell nuclear antigen and MCP-1 (both mRNA and protein). The areas affected by inflammatory changes, as well as the expression of MCP-1 mRNA, declined after longer (28 days) treatment with L-NAME and were replaced by vascular and myocardial remodeling. Our results support the hypothesis that the inhibition of NO synthesis induces inflammatory changes in coronary vascular and myocardial tissues and involves MCP-1 expression. Results also suggest that the early stages of inflammatory changes are important in the development of later-stage structural changes observed in rat hearts. PMID- 9743236 TI - Methylenetetrahydrofolate reductase gene polymorphism and ischemic stroke in Japanese. AB - Hyperhomocyst(e)inemia has been identified as an independent risk factor for atherosclerotic and thromboembolic diseases such as coronary artery disease, cerebral artery disease, and venous thrombosis. Recently, the alanine/valine (A/V) gene polymorphism of 5,10-methylenetetrahydrofolate reductase (MTHFR), one of the key enzymes that catalyzes the remethylation of homocysteine, was reported. The VV genotype is correlated with increased plasma homocyst(e)ine levels as a result of the reduced activity and increased thermolability of this enzyme. In this study, we examined the association between the V allele of the MTHFR gene and ischemic stroke in an elderly Japanese population. The diagnosis of cerebral infarction of all study patients was confirmed by CT of the brain. The MTHFR genotype was analyzed by polymerase chain reaction followed by HinfI digestion. In 256 stroke patients and 325 control subjects, the frequencies of the V allele were 0.45 and 0.32, respectively. The odds ratios and 95% confidence intervals adjusted for the other risk factors were, respectively, 1.51 (1.02 to 2.23) for the AV genotype and 3.35 (1.94 to 5.77) for the VV genotype compared with the AA genotype. Both of these effects were statistically significant (P=0.041 and P<0.001, respectively). In patients with multiple infarcts in particular, the allele frequency of the V mutation was 0.56, and the association between the V allele and stroke was highly significant. Plasma homocyst(e)ine levels were significantly higher in patients with the VV genotype than in patients with the AA or AV genotype, especially those with low plasma folate levels. The V allele of the MTHFR gene was significantly associated with cerebral infarction in an elderly Japanese population in a codominant manner. The VV genotype may contribute to risk for ischemic stroke through a predisposition to increased plasma homocyst(e)ine levels, and dietary folate supplementation may be of benefit, particularly to patients with this genotype. PMID- 9743237 TI - Effects of superoxide anions on endothelial Ca2+ signaling pathways. AB - Although the involvement of free radicals in the development of endothelial dysfunction under pathological conditions, like diabetes and hypercholesterolemia, has been proposed frequently, there is limited knowledge as to how superoxide anions (O2-) might affect endothelial signal transduction. In this study, we investigated the effects of preincubation with the O2(-) generating system xanthine oxidase/hypoxanthine (XO/HX) on mechanisms for Ca2+ signaling in cultured porcine aortic endothelial cells. Incubation of cells with XO/HX yielded increased intracellular Ca2+ release and capacitative Ca2+ entry in response to bradykinin and ATP in a time- and concentration-dependent manner. This effect was prevented by superoxide dismutase but not by the tyrosine kinase inhibitor tyrphostin A48. In addition, capacitative Ca2+ entry induced by the receptor-independent stimulus 2,5-di-(tert-butyl)-1,4-benzohydroquinone or thapsigargin was enhanced in O2(-)-exposed cells (+38% and +32%, respectively). Increased Ca2+ release in response to bradykinin in XO/HX-pretreated cells might be due to enhanced formation of inositol-1,4,5-trisphosphate (+140%). Exposure to XO/HX also affected other signal transduction mechanisms involved in endothelial Ca2+ signaling, such as microsomal cytochrome P450 epoxygenase and membrane hyperpolarization to Ca2+ store depletion with thapsigargin (+103% and +48%, respectively) and tyrosine kinase activity (+97%). A comparison of bradykinin initiated intracellular Ca2+ release and thapsigargin-induced hyperpolarization with membrane viscosity modulated by XO/HX (decrease in viscosity) or cholesterol (increase in viscosity) reflected a negative correlation between bradykinin initiated Ca2+ release and membrane viscosity. Because intracellular Ca2+ is a main regulator of endothelial vascular function, our data suggest that O2- anions are involved in regulation of the vascular endothelium. PMID- 9743239 TI - Dietary fish oils modify the assembly of VLDL and expression of the LDL receptor in rabbit liver. AB - Supplementation of the diet of rabbits with fish oil or sunflower oil resulted in significant changes in the lipoproteins and lipids in serum. Compared with chow fed rabbits, dietary fish oils decreased very low density lipoprotein (VLDL), increased low density lipoprotein (LDL), and shifted the peak of the LDL to denser fractions, whereas sunflower oil increased high density lipoprotein and shifted LDL to the lighter fractions. The amount of LDL receptors in fish oil-fed rabbit liver decreased by > 70% while there was only a small fall in these levels in sunflower oil-fed rabbit liver. The concentrations of apolipoprotein (apo) B in the subcellular organelles of the secretory compartment (rough and smooth endoplasmic reticula and Golgi fractions) were also changed by dietary lipids. In both sunflower oil- and fish oil-fed liver, apo B was increased in the lumen of the rough endoplasmic reticulum compared with fractions from chow-fed rabbit liver. The apo B in the trans-Golgi lumen from fish oil-fed livers was reduced and occurred in particles of d approximately 1.21 g/mL. In contrast, apo B in the trans-Golgi lumen from livers of sunflower oil-fed rabbits was increased and occurred in particles of d < 1.21 g/mL. These results suggests that feeding of fish oils causes an interruption in the intracellular transfer of apo B and hence assembly of VLDL. This leads to an enrichment of the rough endoplasmic reticulum membranes with cholesterol, thus downregulating the expression of the LDL receptor. PMID- 9743238 TI - Hypocoagulant and lipid-lowering effects of dietary n-3 polyunsaturated fatty acids with unchanged platelet activation in rats. AB - We investigated the effects of dietary polyunsaturated fatty acids (PUFAs) on blood lipids and processes that determine hemostatic potential: platelet activation, coagulation, and fibrinolysis. For 8 to 10 weeks, Wistar rats were fed a high-fat diet containing various amounts (2% to 16%) of n-3 PUFAs derived from fish oil (FO) or a diet enriched in n-6 PUFAs from sunflower seed oil (SO). Only the FO diets caused a reduction in mean platelet volume, platelet arachidonate level, and formation of thromboxane B2 by activated platelets, but neither of the diets had a measurable effect on platelet activation. The FO-rich diets decreased the plasma concentrations of triglycerides and cholesterol, whereas the SO diet reduced triglycerides only. Parameters of fibrinolysis and standard coagulation times, ie, activated partial thromboplastin time and prothrombin time, were only marginally influenced by these diets. In contrast, dietary FO, but not SO, led to decreased levels of the vitamin K-dependent coagulation factors prothrombin and factor VII, while the level of antithrombin III was unchanged. The endogenous thrombin potential (ETP) was measured with an assay developed to detect the hypocoagulable state of plasma. After activation with tissue factor and phospholipids, the ETP was reduced by 23% or more in plasma from animals fed a diet with >4% FO. No significant effect of the SO diet on ETP was observed. Control experiments with plasma from warfarin-treated rats indicated that the ETP was more sensitive to changes in prothrombin concentration than in factor VII concentration. Taken together, these results indicate that in rats, prolonged administration of n-3 but not n-6 PUFAs can lead to a hypocoagulable state of plasma through a reduced capacity of vitamin K-dependent thrombin generation, with unchanged thrombin inactivation by antithrombin III. PMID- 9743240 TI - Expression of interleukin-6 in atherosclerotic lesions of male ApoE-knockout mice: inhibition by 17beta-estradiol. AB - Increased levels of interleukin-6 (IL-6) have been proposed to contribute to a number of pathological disorders, including osteoporosis and Alzheimer's disease. In human atherosclerotic lesions, IL-6 protein and mRNA have been detected, although the role of IL-6 in plaque formation is unknown. We have examined the expression pattern of IL-6 mRNA and secreted protein in male apolipoprotein E knockout (apoE-KO) mice aortas. Furthermore, we have evaluated the effects of 17beta-estradiol (E2), a vasculoprotective sex steroid hormone, on the secretion of this inflammatory cytokine from isolated male apoE-KO mice aortas. The expression of IL-6 mRNA was detected by reverse transcription-polymerase chain reaction in the apoE-KO mouse aortas but not in the aortas of age-matched control mice. Similarly, the secretion of IL-6 protein from isolated apoE-KO aortic segments was significantly greater than that from aortas of age-matched control animals. The secretion of IL-6 from isolated aortic rings of apoE-KO mice ranging in age from 6 to 48 weeks showed a significant, positive correlation with percent lesion area measured in the same tissue. Immunohistochemical staining of apoE-KO mouse aortic tissue sections demonstrated colocalization of IL-6 expression with macrophages. Treatment of male apoE-KO mice with E2 for 3 weeks resulted in a statistically significant 50% reduction in IL-6 secretion from ex vivo aortic tissue segments. There was no significant change in total serum cholesterol and triglyceride levels in the E2-treated group compared with placebo-treated controls. These data demonstrate that (1) IL-6 mRNA and protein are expressed in the atherosclerotic plaques of apoE-KO mice aortas and (2) IL-6 production is suppressed by E2 treatment, which may contribute to the antiatherosclerotic effects of E2 in the apoE-KO mouse model of atherosclerosis. PMID- 9743241 TI - Dietary antioxidants inhibit development of fatty streak lesions in the LDL receptor-deficient mouse. AB - Oxidized low density lipoprotein (LDL) promotes atherogenesis. Although pharmacological antioxidants such as probucol inhibit both LDL oxidation and atherosclerosis in hyperlipidemic animals, the effects of natural antioxidants such as vitamin E are inconclusive. To further determine the effects of supplemental dietary antioxidants in vivo, we evaluated whether combined dietary antioxidants (0.1% vitamin E, 0.5% beta-carotene, and 0.05% vitamin C) inhibit LDL oxidation and fatty streak lesion development in homozygous LDL receptor-null (LDLR-/-) mice fed a high-fat, high-cholesterol diet. An additional group of mice were fed black tea, which has been shown to inhibit LDL oxidation in vitro. After receiving a high-fat, high-cholesterol diet for 8 weeks, the combined antioxidant supplemented (antioxidant) group (n=18), tea group (n=19), and control group (n=17) had equivalent plasma cholesterol levels. LDL oxidation, as measured by the lag phase of conjugated diene formation, was markedly inhibited in the antioxidant group compared with the tea or control groups [mean lag phases=143+/ 7 (antioxidant), 100+/-5 (tea), and 84+/-4 (control) minutes; P<0.0001 antioxidant versus tea or control]. The cross-sectional surface area of fatty streak lesions in the aortic sinus was reduced by 60% in the antioxidant group compared with both the tea and control groups (P<0.0001 antioxidant versus tea or control). There was no difference in lesion area between tea and control groups. Although both LDL oxidation and atherosclerosis were significantly inhibited in the antioxidant group, no correlation between lag phase values and lesion size was observed among individual animals. Furthermore, black tea did not inhibit fatty streak development in LDLR-/- mice. These data suggest that combined natural dietary antioxidants inhibit both LDL oxidation and atherogenesis in animals with elevated LDL but that inhibition of LDL oxidation alone may not prevent the development of atherosclerosis. PMID- 9743242 TI - Changes in plasma free and sulfoconjugated catecholamines during the perioperative period of cardiac surgery: effect of continuous infusion of dopamine. AB - In order to elucidate the pharmacological properties of the formation of sulfoconjugated catecholamines (CAs) in human plasma, we investigated the changes in the plasma levels of free and sulfoconjugated CA during the continuous infusion of dopamine (DA; 4-6 microg/kg/min for 24 h, followed by 3-4 microg/kg/min for 48 h) in patients who had undergone cardiac surgery. The plasma level of free DA increased immediately after the start of the infusion and reached a plateau within 1 h at a level of about 2000 times the basal value. In the control patients who had received non-cardiac surgery without DA infusion, plasma-free DA increased only 5-fold after their operation. The plasma level of DA sulfate increased linearly for 24 h, to 48-fold of the basal value by DA infusion, whereas it showed only a 2-fold increase in the control patients. After 24 h, due to reduction of the infused DA dose, the level of free DA gradually decreased, whereas the level of DA sulfate remained elevated. The plasma levels of free adrenaline (Ad) and noradrenaline (NA) also increased during the DA infusion, but their levels reached a plateau within 1-2 h. Sulfoconjugated Ad and NA increased progressively until the tapering off of DA infusion. In the control patients, both free and conjugated Ad and NA showed transient increases over 12 h after surgery. These results suggest that sulfoconjugation plays a role in regulating the plasma levels of excess free CA, thereby modifying the cardiovascular effects of circulating CA. Measurement of the increase in plasma conjugated CA may be useful as an index of the increase in free CA in plasma due to the administration of an exogenous form or release of endogenous CA from the tissues. PMID- 9743243 TI - Endothelium-dependent potentiation of prostaglandin F2alpha-induced contractions by (+/-)-[6]-gingerol is inhibited by cyclooxygenase- but not lipoxygenase inhibitors in mouse mesenteric veins. AB - The mechanism of potentiation of prostaglandin (PG) F2alpha-induced contraction of mouse mesenteric veins by (+/-)-[6-gingerol was investigated in vitro. (+/-) [6]-Gingerol (0.3mM) potentiated the maximal contraction response elicited by PGF2alpha (0.28 mm) in the presence of intact vascular endothelium, but not in its absence (de-endothelialized preparations). The potentiating effect was completely inhibited by cyclooxygenase inhibitors (0.2 mm aspirin and 0.2 mm indomethacin) and partly by calcium antagonists (2 microM verapamil, 8 nM nitrendipine and 1 microM ryanodine), but not inhibited by nordihydroguaiaretic acid (NDGA), a lipoxygenase inhibitor and ONO-3708, a thromboxane (TX) A2 antagonist. The potentiation by (+/-)-[6]-gingerol is also observed in mesenteric veins of streptozotocin-diabetic mice where the enhancement of PGF2alpha-induced contraction is caused mainly by activation of lipoxygenase. The potentiation of PGF2alpha-induced contraction by (+/-)-[6]-gingerol may be caused by a cyclooxygenase-dependent release of vasoconstrictors, other than PGF2alpha and TXA2, or by inhibiting vasorelaxants released from endothelial cells of mouse mesenteric veins. PMID- 9743244 TI - Changes in adrenocorticotropic hormone (ACTH) release from the cultured anterior pituitary cells of streptozotocin-induced diabetic rats. AB - We examined ACTH release from cultured anterior pituitary cells of streptozotocin (STZ)-induced diabetic rats. Rats 1 week after the injection of STZ (55 mg/kg, i.v.) were used as a short-term diabetic model, while rats 8 weeks after the injection of STZ were used as a long-term diabetic model. ACTH release induced by corticotropin-releasing factors (CRF) was significantly increased in the short term diabetic rats, whereas ACTH release decreased in the long-term diabetic rats. A stimulator of adenylate cyclase, forskolin, caused marked increases in ACTH release in short-term diabetic rats, whereas it did not affect the long-term diabetic rats. An L-type Ca2+ channel agonist, BAY K 8644, did not affect ACTH release in the short-term diabetic rats, although it decreased ACTH release in long-term diabetic rats. In addition, CRF-induced ACTH release also increased in normal anterior pituitary cells cultured under high glucose conditions (25 mmol/L) for 5 d. These results suggest that the increase in the CRF-induced ACTH release from cultured anterior pituitary cells of short-term diabetic rats appears to involve the cAMP system in part. In contrast, the decrease in the CRF induced ACTH release from the cultured anterior pituitary cells of long-term diabetic rats may indicate a change in the properties of the L-type Ca2+ channel coupled with the CRF receptor, or in the CRF receptor itself. PMID- 9743245 TI - Endothelin ET(B) receptor-mediated action on systemic and renal hemodynamics and urine formation in deoxycorticosterone acetate-salt-induced hypertensive rats. AB - The pathophysiological role of endothelin ET(B) receptor-mediated action on systemic and renal hemodynamics and urine formation in deoxYcorticosterone acetate (DOCA)-salt hypertensive rats was investigated. An intravenous bolus injection of a selective ET(B) receptor antagonist, BQ788 (1 mg/kg), produced a significant increase in mean arterial pressure (MAP) of DOCA-salt treated rats, whereas the agent-induced increase in MAP was less marked in normotensive sham rats. Administration of BQ788 caused a significant and sustained reduction in renal blood flow both in DOCA-salt and sham rats. No marked effects were observed on urine formation in both groups. Alternatively, a selective ET(A) receptor antagonist, FR139317 (10 mg/kg), produced a potent hypotensive effect, accompanied by significant renal vasodilation in DOCA-salt hypertensive rats, but these effects were partially reversed by the subsequent administration of BQ788. When renal perfusion pressure was protected from FR139317-induced hypotension by an aortic clamp, significant diuresis and natriuresis were observed, events partially reversed by the subsequent administration of BQ788. Our results indicate that the ET(B) receptor-mediated action tonically functions as a hypotensive and a renal vasodilatory factor and that these effects seem to be up regulated in DOCA-salt hypertension. We also suggest that the ET(A) receptor blockade in DOCA-salt hypertensive rats unmasks the ET(B) receptor-mediated action which partially contributes to the antihypertensive effect induced by FR139317. PMID- 9743246 TI - Heparin-selenocystamine conjugate with selenol groups. AB - Heparin-selenocystamine conjugate, which was intended to mimic the heparin selenoprotein P complex, was prepared. The conjugate had glutathione peroxidase like activity and activity was observed toward hydrogen peroxide, tert-butyl hydroperoxide, and cumene hydroperoxide. The ultraviolet spectrum of an aqueous solution of the conjugate was stable and had a similar shape to that observed transiently when selenocystamine was reduced by sodium cyanoborohydride; this suggests that the diselenide bond of selenocystamine introduced into heparin was cleaved during conjugate preparation and the selenol group is preserved. The conjugate reacted to the same degree as cysteine with 5,5'-dithiobis (2 nitrobenzoic acid) (DTNB) releasing thionitro-benzoic acid, which indicated that the selenium in the conjugate is present as selenol. However, the reaction rate of the conjugate was slower than cysteine which may be due to partially restricted access of DTNB to the selenol group in the conjugate. This conjugate had 1,1-diphenyl-2-picryl-hydrazyl(DPPH) radical scavenging activity as well as superoxide anion scavenging activity. These results indicate that the conjugate serves as a useful model compound with a stable selenol group having a range of biological activities, and suggest a possible antioxidant defensive role for the complex of endogenous heparin-like substance and selenoprotein P. PMID- 9743247 TI - Development of bioactive functions in Hydrangeae Dulcis Folium. VII. Immunomodulatory activities of thunberginol A and related compounds on lymphocyte proliferation. AB - Immunomodulatory activities of constituents isolated from Hydrangeae Dulcis Folium and their related compounds on lymphocyte proliferation were investigated using splenocytes and lymph node cells. Thunberginol A (TA) significantly suppressed B lymphocyte proliferation stimulated by lipopolysaccharide (LPS) at 10(-5) M. However TA at a lower concentration (10(-6) M) and the other compounds, except hydrangenol and 3'-hydroxyhydrangeaic acid, tended to potentiate B lymphocytes proliferation (10(-5) M). On the other hand, thunberginol A significantly suppressed T lymphocyte proliferation stimulated by concanavalin A (Con A), but did not suppress phytohemagglutinin (PHA). Hydrocortisone and cyclosporin A strongly suppressed T lymphocyte proliferation induced by both Con A and PHA. These results suggest that thunberginol A acts on both B and T lymphocytes and may have a suppressive mechanism different from the known immunosuppressants. The cytotoxicity of TA for splenocytes seemed weaker than known immunosuppressants resulting from viability tests using MTT assay and the measurement of lactate dehydrogenase (LDH) released in the medium. Moreover, TA suppressed antigen-specific T lymphocyte proliferation in mice lymph node cells immunized with keyhole limpet hemocyanin (KLH). Thus, these findings show that TA has a suppressive effect on lymphocyte activation, and this inhibitory effect of TA seems to contribute to its suppressive effect on type IV allergies. PMID- 9743248 TI - Inhibitory effect of Coptidis Rhizoma and Scutellariae Radix on azoxymethane induced aberrant crypt foci formation in rat colon. AB - This study was conducted to obtain effective cancer chemopreventive agents with low toxicity from medicinal herbs. The effect of aqueous extracts from 9 medicinal herbs with antiinflammatory effect were examined on the formation of azoxymethane (AOM)-induced aberrant crypt foci (ACF), putative preneoplastic lesions of the colon. Male F344 rats were treated with 15 mg/kg body weight of AOM once a week for two weeks. Herbal extract consisting of 2% of the diet was administered from 1 d prior to the first carcinogen treatment. The number of AOM induced ACF per colon was counted at 4 week. Extracts of Coptidis Rhizoma and Scutellariae Radix significantly inhibited AOM-induced ACF formation. The number of ACF was decreased to 54% and 78% of that of the control by 2% Coptidis Rhizoma and Scutellariae Radix extract in the diet, respectively. Berberine and Baicalin, major ingredients of Coptidis Rhizoma and Scutellariae Radix, inhibited ACF formation at a dose equivalent to the amount in each herbal extract. Therefore, to investigate the mechanisms of action of berberine and baicalein which is the active substances of orally administered baicalin, their effects on cyclooxygenase 1 and 2 activities were studied. Berberine was found to inhibit cyclooxygenase 2 activity without inhibition of cyclooxygenase 1 activity, and baicalein inhibited cyclooxygenase 1 activity. Thus, Coptidis Rhizoma and Scutellariae Radix suppressed experimental colon carcinogenesis, and their chemopreventive effects were explained from the inhibition of berberine on cyclooxygenase 2 activity and baicalein on cyclooxygenase 1 activity. PMID- 9743249 TI - Association of liposomes containing a soybean-derived sterylglucoside mixture with rat primary cultured hepatocytes. AB - Dipalmitoylphosphatidylcholine (DPPC) and cholesterol (Ch) liposomes containing a soybean-derived sterylglucoside mixture (SG) (SG-liposomes) are effective for targeting hepatocytes in mice. We investigated uptake of SG-liposomes to hepatocytes compared with that via the galactose receptor. The association of SG liposomes entrapping calcein was examined at 4 degrees C or 37 degrees C, changing incubation time and the SG concentration of the liposomes, and using the inhibitor on the galactose receptor in rat primary cultured hepatocytes. The amount of liposomes recovered with hepatocytes was determined by measuring the concentration of calcein and imaged with confocal laser scanning microscopy in the cultured cells. The association of SG-liposomes at 4 degrees C was significantly decreased compared with that at 37 degrees C as that of liposomes containing lactosylceramide (LC) (LC-liposomes) that were already known to be taken up by hepatocytes via the asialoglycoprotein receptor. While the association of SG-liposomes at both 4 degrees C and 37 degrees C increased with the increase of incubation time, the association of SG-liposomes at 37 degrees C was almost saturated after 1 h. The association of SG-liposomes pretreated with concanavalin A was significantly decreased (to about 40% of that without pretreatment at 37 degrees C) and was at the same level as the association of SG liposomes and non-SG-liposomes at 4 degrees C. The association of the SG liposomes by hepatocytes incubated for I h at 37 degrees C was almost saturated at about 2.0 nmol SG/mg protein. The association of the SG-liposomes with hepatocytes was not inhibited in the presence of LC-liposomes. The affinity of the galactose residue for hepatocytes appeared to be similar to that of the glucose residue in the liposomes, because the amount of sugar residue and the association of SG-liposomes and LC-liposomes were almost the same values. These results suggest that SG-liposomes may be associated with hepatocytes not by a galactose receptor-mediated endocytosis. PMID- 9743250 TI - Antimalarial activity and nucleoside transport inhibitory activity of the triterpenic constituents of Cimicifuga spp. AB - The in vitro antimalarial activity against human malaria parasite (Plasmodium falciparum, FCR-3 strain) was examined using 59 triterpenoids obtained during studies on the triterpenic constituents of Cimicifuga spp. The 50% effective concentration values (EC50) of 25 active triterpenoids were 1.0-3.0 microM, and 19 of the compounds had a common 16, 23:23, 26:24, 25-triepoxy group in the side chain moieties. Among the active triterpenoids, 9 also showed significant inhibition of nucleoside transport in mouse splenocytes. A relationship between the antimalarial activity and the inhibition of nucleoside transport involving these triterpenoids is discussed. PMID- 9743251 TI - Anti-HIV-1 activity of herbs in Labiatae. AB - The anti-HIV-1 activity of aromatic herbs in Labiatae was evaluated in vitro. Forty five extract from among 51 samples obtained from 46 herb species showed significant inhibitory effects against HIV-1 induced cytopathogenicity in MT-4 cells. In particular, the aqueous extracts of Melissa officinalis, a family of Mentha x piperita "grapefruit mint," Mentha x piperita var. crispa, Ocimum basilicum cv "cinnamon," Perilla frutescens var. crispa f. viridis, Prunella vulgaris subsp. asiatica and Satureja montana showed potent anti-HIV-1 activity (with an ED of 16 microg/ml). The active components in the extract samples were found to be water-soluble polar substances, not nonpolar compounds such as essential oils. In addition, these aqueous extracts inhibited giant cell formation in co-culture of Molt-4 cells with and without HIV-1 infection and showed inhibitory activity against HIV-1 reverse transcriptase. PMID- 9743252 TI - Anti-tumor-promoting activity of majonoside-R2 from Vietnamese ginseng, Panax vietnamensis Ha et Grushv. (I). AB - Seven saponins (1-7) isolated from the rhizomes and roots of Panax vietnamensis were tested for their inhibitory effects on Epstein-Barr virus early antigen (EBV EA) induced by the tumor promoter, 12-O-tetradecanoylphorbol-13-acetate (TPA), in Raji cells as a primary screening test for anti-tumor-promoters (cancer chemopreventive agents). The ocotillol-type saponin, majonoside-R2 (2), which is the major and characteristic constituent of this plant, exhibited a significant inhibitory effect on EBV-EA activation. Furthermore, the cell cycle analysis of 2 on Raji cells was also examined and strong inhibition was observed on the effect of the cell cycle induced by TPA. Compound 2 showed potent anti-tumor-promoting activity in two-stage carcinogenesis tests of mouse skin using 7,12 dimethylbenz[a]anthracene (DMBA) as an initiator and TPA or fumonisin B1 as a promoter. Consequently, these results suggest that majonoside-R2 (2) could be a valuable chemopreventive agent against chemical carcinogenesis. PMID- 9743253 TI - Influence of verapamil and digoxin on the in vitro binding of doxorubicin to the rat heart. AB - A study has been carried out to characterize the binding of doxorubicin to heart homogenates and subcellular fractions. The technique chosen to perform the study was equilibrium dialysis and the levels of anthracycline in the samples obtained from the dialysis were assessed using HPLC. Doxorubicin has high affinity to heart homogenates and subcellular fractions such as nuclear, mitochondrial and microsomal. The binding was saturable and dose-dependent. Doxorubicin binding is decreased in the presence of digoxin and especially verapamil. PMID- 9743254 TI - Population pharmacokinetic analysis of mexiletine in adult arrhythmic patients in Japanese population. AB - We analyzed mexiletine in Japanese patients using population pharmacokinetics. 139 serum concentration data, which were collected for therapeutic drug monitoring from 121 patients, were used for this analysis. We also investigated influence of age and gender on pharmacokinetics, and age was found to be an influential factor on clearance. The final pharmacokinetic parameters are, CL/F=0.580-0.00369 x AGE (l/h/kg) and Vd/F=6.63 (l/kg). These results should be useful for adjusting the dosage to a patient's age for the prevention of an adverse reaction caused by overexposure. PMID- 9743255 TI - Development of a computer program, MDGP, for population pharmacokinetic analysis written using ANSI C language on wide platforms. AB - MDGP, a computer program for population pharmacokinetics, has been developed. This program is based on maximum likelihood estimation with poly-dimensional normal distribution errors, and variable metric methods, direction set methods, conjugate gradient methods and the polytope method are provided as minimizations of objective function. The source codes of MDGP are described using ANSI C language. MDGP is able to run on wide platforms equipped with an ANSI C compiler. Users can use algebraic, ordinary differential and Laplace-transformed equations as descriptions of a pharmacokinetic model. Partial differential equations for each model parameter are not needed. Comparison of the estimated parameters, the iteration counts until convergence and objective function value, using MDGP and NONMEM, revealed that the calculation performance of the two programs are comparable. The MDGP is thought to be useful for the analysis of various pharmacokinetic models, including population pharmacokinetic analysis. PMID- 9743256 TI - Enhanced rectal absorption of amphotericin B lyophilized with glycyrrhizinate in rabbits. AB - The influence of bases and additives in the formulation for rectal absorption of amphotericin B (AMB) lyophilized with dipotassium glycyrrhizinate (GLYK) was investigated using rabbits in relation to an in vitro release test. The release of AMB from the fatty base of Witepsol or a medium chain triglyceride (MCT) was markedly faster than that from the hydrophilic base of macrogol. The addition of polyoxyethylene (2) lauryl ether (POE(2)LE) into the fatty bases led to a marked increase in the release rate, whereas POE(9)LE or sodium lauryl sulfate resulted in a significantly lower release rate. Animals received rectally each of seven AMB formulations of Witepsol H-15, macrogol, MCT with surfactants and aqueous solution. The absorption of the AMB lyophilized mixture with GLYK at a 1:9 molar ratio from a MCT base was significantly superior to that from macrogol. The addition of POE(2)LE into the MCT base resulted in a marked increase in bioavailability, showing the highest bioavailability of 4.9%. High serum levels of over 100 ng/ml of serum were maintained for 24 h following administration. The lowest bioavailability was 0.32% for the macrogol suppository. There was a good correlation between the release rate of AMB from the formulations and bioavailability. These results suggest that an AMB rectal formulation may provide a promising therapeutic alternative to infusion, taking into account the serum level of AMB exceeding the minimal inhibitory concentration of the infecting organism. PMID- 9743257 TI - Uptake of basic drugs into rat lung granule fraction in vitro. AB - Although basic drugs are distributed widely in various tissues, they are characteristically concentrated in the lung granule fraction. We examined the uptake of seven lipophilic basic drugs into rat lung granule fraction (P2) in vitro and investigated the contributions of drug lipophilicity and lysosomal trapping to the characteristic lung P2 distribution. The uptake of each drug into P2 was examined at various pH values. The drug concentration in P2 was determined by gas chromatography. Biperiden (BP) was rapidly taken up into P2, reaching a maximal concentration within 1 min at pH 7.4 at both 4 degrees C and 37 degrees C. Both BP and chlorpromazine uptake into P2 was biphasic. Though the uptake rates of the seven drugs into P2 increased with rising pH, the rate of increase varied for each drug. There was a good correlation between the octanol-water partition coefficient of the non-ionized form (P(oct)) of each drug and the uptake into P2 in the presence or absence of NH4Cl, which inhibits lysosomal trapping. However, uptake into P2 in the presence of NH4Cl showed a stronger P(oct)-dependency. We conclude that the distribution of basic drugs into lung P2 is dependent on both drug lipophilicity and lysosomal uptake. PMID- 9743258 TI - Nasal insulin delivery in rabbits using soybean-derived sterylglucoside and sterol mixtures as novel enhancers in suspension dosage forms. AB - The effect of a soybean-derived sterol mixture (SS) and a steryl glucoside mixture (SG) as enhancers of the nasal absorption of insulin in rabbits was investigated. SS consists of beta-sitosterol (Sit), campesterol (Camp), stigmasterol (Stig) and brassicasterol (Bras), and SG is a mixture of their monoglucosides. For each component of SS tested for efficacy in promoting the systemic absorption of nasally administered insulin, the following order was observed: Sit> or =Camp>Stig. This finding was in agreement with the order of the enhancers' lipophilicity. In the case of SG, the effect of beta-sitosterol beta-D glucoside (Sit-G) was significantly greater than that of SG. The pharmacological bioavailability was 6.7% for SG and 11.3% for Sit-G in the suspension dosage forms. SG showed a greater degree of enhancement of insulin permeation through the nasal mucosa than SS. To elucidate the contribution of SG to the enhanced absorption, insulin permeation through an artificial membrane and the nasal mucosa was investigated in vitro, and the results were compared with those for SS. The findings suggest that SG and SS have some effect on nasal mucosa lipids. PMID- 9743259 TI - High absorbency and subchronic morphologic effects on the nasal epithelium of a nasal insulin powder dosage form with a soybean-derived sterylglucoside mixture in rabbits. AB - A soybean-derived sterylglucoside mixture (SG) is a potential enhancer of the nasal absorption of insulin. The aim of the study was to examine the absorption of insulin given as the powder dosage form with SG and excipient and to determine the subchronic effects of SG on the morphology of rabbit nasal epithelium. The insulin powder dosage form with SG was administered to the rabbit nasal cavity for five successive days. The average bioavailability and the average pharmacological bioavailability of insulin were about 25.0 and 61.6%, respectively. The nasal mucosa was taken from the nasal cavity and side-effects were investigated using an optiphoto light microscope. The insulin powder dosage form with SG produced no signs of inflammation, erosion or squamous metaplasia. These findings indicate that SG can be considered as a safe and effective enhancer and excipient in the nasal insulin powder dosage form. PMID- 9743260 TI - First-pass metabolism of 5-fluorouracil in the perfused rat small intestine. AB - The first-pass intestinal metabolism of 5-fluorouracil (5-FU) was investigated by single-pass perfusion of the rat small intestine. At the low concentration of 0.06 mg/ml, the fraction of 5-FU absorbed into (i.e., appeared in) the mesenteric venous blood (Fa,b) was about 50% smaller than the fraction absorbed (disappeared) from the intestinal lumen (Fa), indicating the first-pass extraction of 5-FU in the intestinal mucosa. By addition of uracil (6 mg/ml), the Fa of 5-FU was reduced presumably by competition for the pyrimidine carrier at the process of intestinal uptake (entry into the mucosa). The Fa,b was also reduced, but to a lesser extent, resulting in insignificant first-pass extraction. These results suggest that the extraction of 5-FU in the absence of uracil is caused by metabolism and that uracil is a competitor for this pathway. When 5-FU concentration was raised from 0.06 to 0.6 mg/ml in the absence of uracil, the Fa was reduced by about 50%, consistent with the suggestion of the involvement of saturable uptake by the pyrimidine carrier, and thereafter remained unchanged at 6 mg/ml. However, since Fa,b was also reduced by a similar extent, the intestinal availability (FI=Fa,b/Fa) was unchanged at about 0.5, indicating that the intestinal first-pass extraction of 5-FU is independent of concentration with the extraction ratio (difference between unity and FI) of about 0.5 over the wide range of concentration from 0.06 to 6 mg/ml. Thus, the present study demonstrates that the significant first-pass metabolic extraction of 5-FU occurs in the mucosa of the small intestine, supporting our previous suggestion that 5-FU undergoes first-pass metabolism not only in the liver but also in the small intestine after oral administration. Considering that the oral bioavailability of 5-FU in the human (28%) is reportedly comparable with that in the rat (28%), it is likely that intestinal first-pass metabolism may be significant also in the human. Intestinal first-pass metabolism should be taken into account to explore more efficient and controlled oral 5-FU therapy. PMID- 9743261 TI - Characterization of the transport of a cationic octapeptide, octreotide, in rat bile canalicular membrane: possible involvement of P-glycoprotein. AB - The cyclic cationic octapeptide octreotide is known to be taken up by hepatocytes, with more than 70% of an intravenous dose being excreted into bile in unchanged form. We have already reported that a transporter other than the canalicular multispecific organic anion transporter (cMOAT) is responsible for the biliary excretion of octreotide in vivo. The aim of this study is to obtain an insight into the transporter of octreotide in bile canalicular membrane. The effect of various compounds on the ATP-dependent uptake of octreotide by rat bile canalicular membrane vesicles (CMV) was investigated. The ATP-dependent uptake of [14C]octreotide by CMV was inhibited by verapamil, vincristine and PSC833 in a concentration-dependent manner, the maximum inhibitory effects of these compounds being almost equal to that of excess unlabeled octreotide, while taurocholic acid (TCA) caused no inhibition at concentrations much higher than the Km of TCA uptake by CMV. Mutual inhibition between octreotide and dinitrophenylglutathione (DNP-SG), a representative substrate for cMOAT was only minor and could only be observed at concentrations much higher than the Km for each ligand uptake. To examine the contribution of P-glycoprotein to the biliary excretion of octreotide in vivo, biliary excretion of octreotide was compared between P-glycoprotein induced rats by phenothiazine (PTZ) treatment and normal rats. A significant increase in the biliary excretion rate was observed in PTZ-treated rats. Only a slight decrease in biliary excretion was observed in mdr1a knock-out mice compared with normal mice, which may be explained by the associated induction of mdr1b. These results demonstrate that the transporter for octreotide is different from cMOAT and the bile acid transporter. The involvement of P-glycoprotein in the biliary excretion of octreotide is suggested. PMID- 9743262 TI - Properties and tissue distribution of mouse monomeric carbonyl reductase. AB - We previously cloned a cDNA for mouse cerebellum carbonyl reductase which shows more than 81% homology to the cDNAs for monomeric carbonyl reductases of the rat, rabbit and human, and for pig 20beta-hydroxysteroid dehydrogenase. In the present study, we expressed the recombinant monomeric enzyme (34 kDa and pI 8.3) from the cDNA and compared its properties with the recombinant human enzyme. The mouse and human enzymes showed similar functional properties, although they differed in kinetic constants for carbonyl substrates and in inhibitor sensitivity. Both enzymes lacked glutathione S-transferase activity. Western blot and reverse transcription-polymerase chain reaction analyses showed that the enzyme protein and its mRNA are expressed in various mouse tissues. PMID- 9743263 TI - Evaluation of the topical delivery of a prednisolone derivative based upon percutaneous penetration kinetic analysis. AB - Prednisolone (PN) and an esterified derivative (PND) were evaluated in pharmacological and pharmacokinetic studies. The pharmacological study was performed using a rat croton-oil induced ear edema model. The results for the topical effect in skin and the systemic effect through multiple topical applications showed that PN and PND were equally potent in suppressing edema, and that PN caused a reduction in thymus weight, whereas PND had little effect. The concentration of these steroids in hairless mouse skin was estimated from an in vitro percutaneous absorption study using the computer simulation program MULTI(FILT). PND was found to be poorly absorbed. In fact, the PND concentration in the viable skin remained low (0.79 microg/cm2), even after 7 d. However, the estimated concentration of PND in the viable skin appears to be in excess of the threshold for effective topical effect during the pharmacological evaluation. In contrast, in the case of PN, the estimated PN concentration increased gradually after application and reached a level of 10.22 microg/cm2 at day 7, suggesting that this increase in PN concentration in the viable skin could result in a systemic effect. The difference between PN and PND concentration in the skin during the time course could be due to the metabolism of PND to PN in the viable skin. Consequently, the difference between the pharmacological study is reflected from the results of the pharmacokinetics of PN and PND in the skin. PMID- 9743264 TI - Trapping of secreted molecules at single-cell level using an HPLC resin particle under a videomicroscope. AB - Particles of HPLC resins are used for the trapping of secreted molecules from a single cell. The basic molecules, e.g., histamine, are secreted from a single RBL 2H3 cell by granule exocytosis and are trapped by cation-exchange HPLC resins outside the cell. Since quinacrine is concentrated into the exocytotic and acidic microgranules in RBL-2H3 cells, which are used as a model cell line of mast cells, we measured the change in the fluorescence intensity of the quinacrine released from the cells and that of molecules trapped on the resin using a videomicroscope. By measuring the increase in the fluorescence intensity of the resins, we can estimate the real time course of molecular secretion from a single cell. PMID- 9743265 TI - Interictal inhibitory mechanisms in patients with cryptogenic motor cortex epilepsy: a study of the silent period following transcranial magnetic stimulation. AB - The silent period (SP) following transcranial magnetic stimulation (TMS) of the motor cortex is mainly due to cortical inhibitory mechanisms. The aim of the present study was to investigate these inhibitory phenomena in primary motor cortex epilepsy. We studied the TMS-induced SP in both the first dorsal interosseous (FDI) muscles in 8 patients who suffered from cryptogenic partial epilepsy with seizures starting with clonic movements of the right upper limb. All patients were on chronic medication with antiepileptic drugs. Therefore, besides contrasting the results with 16 age-matched normal controls, we also studied 10 patients receiving similar antiepileptic treatments who suffered from cryptogenic partial epilepsy with seizures characterised by the absence of clonic manifestations. The duration of the SP was bilaterally increased in the patients with clonic seizures when compared with the two other groups of subjects. The SP was longer in the left FDI muscle (contralateral to the side of the clonic manifestation in all the patients). Our findings likely indicate enhanced interictal inhibitory mechanisms in patients with partial epilepsy involving the primary motor cortex. The resulting inhibitory effect could be greater in the intact hemisphere rather than in the affected one, in which the hyperexcitability of the epileptic focus had to be counterbalanced. PMID- 9743266 TI - The photic driving EEG response and photoreactive cerebral blood flow in the posterior cerebral artery in controls and in patients with epilepsy. AB - OBJECTIVES: Instantaneous changes in blood flow velocities during visual stimulation can be assessed by transcranial Doppler sonography (TCD). METHODS: We investigated the possible relationship between the characteristics of photic driving in the EEG elicited by repetitive intermittent photic stimulation and the photoreactive flow changes in the posterior and middle cerebral artery (PCA, MCA) of 25 normal controls and 25 patients with focal epilepsy. Cerebral blood flow velocities (CBFV) of the right PCA (P2 segment) and the left middle cerebral artery (MCA) were measured using a 2 Hz transcranial Doppler device. Simultaneously, scalp EEGs were recorded. RESULTS: During photic stimulation the mean CBFV increase was 20.4 +/- 9.5% in the PCA of the controls (n = 132 stimulations) and 16.0 +/- 10.8% in epileptic patients (n = 150 stimulations, P < 0.01). During those stimulation series with a good EEG driving response (n = 203), the mean increase of CBFV in the PCA was 19.7 +/- 10.0%, as opposed to 14.4 +/- 10.5% during the stimulations with a poor EEG response (n = 79, P < 0.01). A good photic driving response was associated with a higher increase of CBFV in the PCA than a poor one. The increase in CBFV of the PCA in normal controls was higher than in patients with focal epilepsy. CONCLUSIONS: This may indicate that epileptic patients have a reduced coupling between neuronal activation and blood flow. PMID- 9743267 TI - Responses to photic stimulation in patients with occipital spikes. AB - OBJECTIVE: To determine the effect of intermittent photic stimulation (IPS) and frequency of asymmetric driving responses in patients with occipital spikes. METHODS: The amplitude of the driving response at 4 flash frequencies was measured from a referential montage in 60 patients with occipital spikes and in 60 normal EEG records from age-matched patients. Responses were classified as asymmetric if the amplitude at one occipital area was less than 50% of the amplitude at the other. RESULTS: A measurable photic response occurred significantly less frequently in patients with occipital spikes (48%) compared to the control group (70%; Fisher's test P < 0.05). The driving response was asymmetric in 7/36 patients (37%) with unilateral spike foci versus none in the control group (Fisher's test, P < 0.001). The amplitude was suppressed ipsilateral to the focus in 5 patients, all of whom had an ipsilateral structural lesion or focal slowing. In two cases the amplitude was higher ipsilateral to the focus, neither having slowing or a structural lesion. CONCLUSIONS: Patients with occipital spikes have an increased frequency of asymmetric driving response. An attenuated response ipsilateral to the focus seems to be related to an underlying lesion while the presence of an epileptiform focus in some cases with no slowing on EEG and normal imaging studies may lead to an accentuation of this response. PMID- 9743268 TI - Scalp topography and source analysis of interictal spontaneous spikes and evoked spikes by digital stimulation in benign rolandic epilepsy. AB - OBJECTIVES: We report the analysis of scalp topography and dipole modeling of the rolandic spikes in 6 patients suffering of benign rolandic epilepsy of childhood with extremely high amplitude SEP by tapping stimulation of the finger of the hand. METHODS: EEG and BESA analysis were performed for both rolandic spontaneous interictal spikes and high amplitude scalp activity evoked by tapping and electrical stimulation of the first finger of the right hand. RESULTS: The evoked responses showed a morphology characterized by a rapid phase (spike) followed by a slow phase (slow wave). The spike presented an early small positive component followed by a main negative component. Similar morphology, dipole configuration and source localization were observed for both rolandic spikes and evoked high amplitude scalp responses. Dipole localization showed an overlap of spatial coordinates between rolandic and evoked spikes. CONCLUSIONS: These findings suggest that the extremely high amplitude SEPs could be evoked spikes which probably had the same cortical generators of the spontaneous rolandic spikes. PMID- 9743269 TI - Seizure detection using a self-organizing neural network: validation and comparison with other detection strategies. AB - OBJECTIVE: A previously described seizure detection algorithm (CNET) (Gabor, A.J., Leach, R.R. and Dowla, F.U. Automated seizure detection using a self organizing neural network. Electroenceph. clin. Neurophysiol., 1996, 99: 257-266) was validated with 200 records from 65 patients (4553.8 h of recording) containing 181 seizures. DESIGN AND METHODS: Performance of the algorithm was manifest by its sensitivity ((seizures detected/total seizures) x 100) and selectivity (false-positive errors/Hr-FPH). Comparisons with the Monitor detection algorithm (Version 8.0c, Stellate Systems) and audio-transformation (Oxford Medilog) were performed. RESULTS: CNET detected 92.8% of the seizures and had a mean FPH of 1.35 +/- 1.35. Monitor detected 74.4% of the seizures and had a mean FPH of 3.02 +/- 2.78. Audio-transformation detected all but 3 (98.3%) of the seizures. Selectivity for this detection strategy was not defined. CONCLUSIONS: This study not only validates the CNET algorithm, but also the notion that seizures have frequency-amplitude features that are localized in signal space and can be selectively identified as being distinct from other types of EEG patterns. The ear is a specialized frequency-amplitude detector and when the signal is transformed into audio frequency range (audio-transformation), seizures can be detected with better sensitivity as compared to the other strategies examined. PMID- 9743270 TI - Sleep latency on the maintenance of wakefulness test (MWT) for 530 patients with narcolepsy while free of psychoactive drugs. AB - OBJECTIVES: To compare maintenance of wakefulness test (MWT) data gathered at baseline in the course of two, multicenter studies on the therapeutic efficacy of modafinil with published MWT norms. METHODS: The MWT is a procedure that uses electrophysiological measures to determine the ability to remain awake while sitting in a quiet, darkened room. The test consists of 4 20 min trials conducted 4 times at 2 h intervals commencing 2 h after awakening from a night of sleep. MWT data were gathered at baseline in the course of two, multicenter studies on the therapeutic efficacy of modafinil. Subjects were 17-68 year old men (n = 239) and women (n = 291) diagnosed with narcolepsy according to the International Classification of Sleep Disorders (ICSD). All patients were free of psychoactive medication for a minimum of 14 days. RESULTS: Mean MWT sleep latency was 6.0 +/- 4.8 min. However, the mean for the first MWT trial was 7.0 min which was longer that the means for the following 3 trials (5.8, 5.6 and 5.7 min, respectively). The 4 distributions of the individual MWT trials were similar and adequately summarized by the distribution of the average MWT sleep latency. As a group, patients with narcolepsy were less able to remain awake than normals; only 8 of 530 (1.5%) patients were able to remain awake on 4 20 min MWT trials compared with 35 of 64 (54.7%) normals in another study. However, using a mean MWT sleep latency of 12 min (the 5th percentile for normals) as the lowest cut-point for normalcy, 15% of patients with narcolepsy appeared to have an unimpaired ability to remain awake. CONCLUSIONS: The diagnosis of narcolepsy did not always predict inability to remain awake on the MWT. Age, gender and the duration of illness did not predict ability to remain awake. Patients with severe cataplexy and other ancillary symptoms were least able to remain awake on MWT trials. Patients who used tobacco and caffeine moderately had the lowest MWT sleep latencies relative to patients with heavy and light use. PMID- 9743271 TI - EEG changes in tuberculous meningitis: a clinicoradiological correlation. AB - OBJECTIVES: The present study is aimed at describing electroencephalographic (EEG) changes in the tubercular meningitis (TBM) and correlating these with clinical and radiological findings. METHODS: All the patients underwent a detailed neurological evaluation, CSF analysis, EEG and CT scan studies. Outcome was assessed by the Barthel index (BI) score at the end of 3 months, into good (BI > or = 12) and poor (BI < 12). Thirty-two patients with TBM have been included of which 3 were definite and the remaining highly probable. Their mean age was 28 (range 8-62) years and 8 of whom were females. The majority of these patients were in stage III. RESULTS: Clinical signs of raised intracranial tension were present in 9 and history of seizure in 11 patients. Cranial CT scan was abnormal in 22 patients. The CT scan abnormalities included hydrocephalus in 20, infarction in 11, exudates in 7 and tuberculoma in 4 patients. The EEG was abnormal in 24 patients. The EEG abnormalities included diffuse theta to delta slowing in 22 patients, intermittent rhythmic delta activity in frontal region in 15, right to left asymmetry in 5 and epileptiform discharges in 4 patients. At the end of 3 months, 5 patients died, 13 had poor, 3 partial and 11 complete recovery. The EEG findings correlated with the severity of meningitis, the degree of coma and outcome at 3 months as assessed by Barthel index score. PMID- 9743272 TI - Automatic EEG analysis during long-term monitoring in the ICU. AB - To assist in the reviewing of prolonged EEGs, we have developed an automatic EEG analysis method that can be used to compress the prolonged EEG into two pages. The proposed approach of Automatic Analysis of Segmented-EEG (AAS-EEG) consists of 4 basic steps: (1) segmentation; (2) feature extraction; (3) classification; and (4) presentation. The idea is to break down the EEG into stationary segments and extract features that can be used to classify the segments into groups of like patterns. The final step involves the presentation of the processed data in a compressed form. This is done by providing the EEGer with a representative sample from each group of EEG patterns and a compressed time profile of the complete EEG. To verify the above approach, 41 6 h EEG records were assessed for normality via the AAS-EEG and conventional EEG approaches. The difference between the overall assessment via compressed and conventional EEG was within one abnormality level 100% of the time, and within one-half level for 73.6% of the records. We demonstrated the feasibility and reliability of automatically segmenting and clustering the EEG, thus allowing the reduction of a 6 h tracing to a few representative segments and their time sequence. This should facilitate review of long recordings during monitoring in the ICU. PMID- 9743273 TI - Reappraisal of the effect of electrode property on recording slow potentials. AB - Subdural electrodes made of stainless steel, which were believed to be unsuitable for recording slow potentials, can still record Bereitschaftspotential (BP) (Neshige, R., Luders, H. and Shibasaki, H. Recording of movement-related potentials from scalp and cortex in man. Brain, 1988, 11: 719-736) and ictal DC shifts (Ikeda, A., Terada, K., Mikuni, N., Burgess, R.C., Comair, Y., Taki, W., Hamano, T., Kimura, J., Luders, H.O. and Shibasaki, H. Subdural recording of ictal DC shifts in neocortical seizures in humans. Epilepsia, 1996b, 37: 662-674) sufficiently. In this study, therefore, the effects of different kinds of metals on slow potential recordings were reevaluated. First, slow electro-oculograms (EOGs) were recorded with 3 different levels of input impedance (200 M omega, 470 k omega and 10 k omega) of a DC amplifier by using surface electrodes made of silver (Ag), silver/silver chloride (Ag/AgCl) and stainless steel. Secondly, BP was recorded by using the above electrodes with a long time constant of 3 s and with a fixed input impedance of 100 M omega. As a result: (1) slow EOGs were equally recorded with the input impedance of 200 M omega and 470 k omega regardless of the kind of metals used, although stainless steel electrodes caused baseline fluctuation, (2) low input impedance of 10 k omega allowed only the Ag/AgCl electrode to record slow EOGs without any decay, and (3) electrodes made of stainless steel could record BP as efficiently as the other two types of electrode with high input impedance. In conclusion, electrodes with a large surface area contact such as cup electrodes and an amplifier with a large input impedance, electrodes made of Ag, and even of stainless steel, can record slow potentials reasonably well. PMID- 9743274 TI - Effect of subthreshold target stimuli on event-related potentials. AB - OBJECTIVES: Event-related potentials (ERPs) elicited by subthreshold visual stimuli were studied to assess the relationship between unconscious cognitive processing and the electrical activity of the brain. METHODS: A new method of modified visual oddball paradigm with supraliminal and subliminal stimuli was applied. Prior to the experiment, the individual 'subjective' threshold for the conscious discrimination between frequent and target stimuli was established for each subject. Supraliminal and subliminal, frequent and target visual stimuli were then alternatively presented in random order to each subject. RESULTS: Both the individual and the grand average ERPs revealed a typical response (P3) in the parietal region after supraliminally presented target stimuli. In subliminal conditions an analogous positive deflection in the central-parietal region was observed, which was elicited by the target stimulus, but not the frequent stimulus. Its latency could be clearly distinguished from the latency of the classical P3, the time difference between the two waveforms was approximately 100 ms. RESULTS: The results of this study revealed the impact of unconscious processing to target stimuli on the configuration of event-related responses. PMID- 9743275 TI - Clinical applications of motor evoked potentials. AB - Magnetic stimulation of brain and spinal roots provides a non-invasive evaluation of nervous propagation as well as of motor cortex excitability in healthy subjects and in patients affected by neurological diseases (i.e. multiple sclerosis, stroke, Parkinson's disease, myelopathies etc.). Motor areas can be reliably mapped and short- and long-term 'plastic' changes of neural connections can be studied and monitored over time. By evaluating excitatory and inhibitory phenomena following transcranial stimuli, the mechanisms of action of different drugs, including antiepileptics, can be studied. Moreover, transcranial stimulation of non-motor brain areas represents a probe for the evaluation of lateralized hemispheric properties connected with higher cortical functions. Recent studies suggest a therapeutic role of repetitive magnetic stimulation in psychiatric disorders. PMID- 9743276 TI - Post-movement beta oscillations studied with linear estimation. AB - The application of surface laplacian and linear estimation methods to single trial EEG data was studied. EEG was recorded in 3 subjects during voluntary, self paced extensions and flexions of the index finger. In each subject a post movement beta synchronisation was found in specific frequency bands. The surface laplacian estimates were calculated using spherical splines and cortical current distributions were constructed using the linear estimation method. Both methods yield similar results and reveal a maximal event-related synchronisation over the left sensorimotor area approximately 500-750 ms after termination of movement. PMID- 9743277 TI - The Pittsburgh study of normal sleep in young adults: focus on the relationship between waking and sleeping EEG spectral patterns. AB - The effects of age and gender on spectral characteristics of the waking EEG were investigated in a large sample of young adult men and women. In addition, relationships between spectral characteristics of the waking and sleeping EEG within an individual were explored. The sample included 28 females and 33 males in two age groups: 20-29 years (n = 32), and 30-40 years (n = 29). Spectral analysis was used to quantify EEG frequency characteristics for waking EEG just prior to sleep onset, as well as for the entire sleep recording. Significant effects of age were seen in the waking EEG but only in the delta frequency range (0.5-4.5 Hz) with lower delta activity in the older group (F = 11.6, P = 0.001). No significant gender effects were found in the waking EEG. Independent of age and gender, spectral profiles in the delta, theta, alpha and beta frequency bands of a subject's waking EEG were found to be highly correlated with their sleep EEG. In addition, subjects with high voltage alpha profiles during waking were found to sleep significantly longer and deeper than those with low voltage records. Significant correlations between waking and sleep EEG suggest that the spectral signature of an individual's EEG may be found across sleep/wake states. PMID- 9743278 TI - Scoring of sleep and wakefulness by behavioral analysis from video recordings in rhesus monkeys: comparison with conventional EEG analysis. AB - Extensive work on sleep-wake cycles in non-human primates has been carried out using conventional EEG scoring. In this study, simultaneous somnopolygrams and video recordings at 1 frame/s were performed on 6 adult rhesus monkeys (Macaca mulatta) during a 24 h period. Wakefulness, NREM sleep and REM sleep were scored by analysis of animal behavior from video data, using characteristic criteria for each state of vigilance. Results were then compared with those of conventional EEG scoring. Values of the total amount for each state obtained by the two scoring methods during the light and the dark periods were significantly closely related (P < 0.001) with a high correlation coefficient for wakefulness (r1 = 0.99956), for NREM sleep (r1 = 0.99641) and for REM sleep (r1 = 0.98708). Moreover, the epoch by epoch analysis between both methods showed a high concordance with percent agreement values of 95.68% for wakefulness, 93.52% for NREM sleep and 94.02% for REM sleep. The number of REM sleep episodes was similarly defined. The patterns of successive sleep-wake cycles determined from both scorings were superimposable, as were the frequent state changes for the same time segments. The video method's main limitation was that the 4 stages of NREM sleep could not be differentiated. Reliability and advantages of sleep-wake scoring by behavioral analysis are discussed. These results suggest that the video methodology is relevant as a non-invasive technique complementary to conventional EEG analysis for sleep studies in rhesus monkeys. PMID- 9743279 TI - Quantitative electro-oculography and electroencephalography as indices of alertness. AB - Even though both electro-oculography (EOG) and EEG have been widely used for assessing alertness, the relationship between these two measures has not yet been clarified at various alertness levels. We estimated the frequencies of eye movements faster than 15 degrees/s (NoEM15) and EEG power at every 0.5 Hz step frequency point, quantitatively as well as continuously from alert to very light sleep (Stage 1) in 14 healthy adults. We devised a new EOG derivation for a computerized analysis, because conventional EOG recordings suffer from EEG contamination and have been analyzed manually. EOG electrodes were attached to the left orbitale in this study. The eye movements were detected as peaks in differentiated EOG signals. Significant correlations were found between NoEM15 and EEG powers at several frequency points, mainly in the alpha and beta bands. Though fluctuating EEG components in close correspondence to the frequency of the eye movements varied depending on the individual, EEG power at the peak frequency of the awake state was most closely associated with the eye movements. A broad band power that centers at the peak frequency may be a better measure for alertness assessment than powers in fixed bands. PMID- 9743280 TI - Non-linear dynamical analysis of the EEG in Alzheimer's disease with optimal embedding dimension. AB - We used non-linear analysis to investigate the dynamical properties underlying the EEG in patients with Alzheimer's disease. We calculated the correlation dimension D2 and the first positive Lyapunov exponent L1. We employed a new method, which was proposed by Kennel et al., to calculate the non-linear invariant measures. That method determined the proper minimum embedding dimension by looking at the behavior of nearest neighbors under a change in the embedding dimension d from d to d + 1. We demonstrated that for limited noisy data, our algorithm was strikingly faster and more accurate than previous ones. Also, we found that, in almost all channels, patients with Alzheimer's disease had significantly lower D2 and L1 values than those for age-approximated healthy controls. These results suggest that brains afflicted by Alzheimer's disease show behaviors which are less chaotic than those of normal healthy brains. In this paper, we show that non-linear analysis can provide a fruitful tool for detecting relative changes, which cannot be detected by conventional linear analysis, in the complexity of brain dynamics. We propose that non-linear dynamical analyses of the EEGs from patients with Alzheimer's disease will be a diagnostic modality in the appropriate clinical setting. PMID- 9743281 TI - EEG coherence in Alzheimer's disease. AB - EEG coherence can be used to evaluate the functionality of cortical connections and to get information about the synchronization of the regional cortical activity. We studied EEG coherence in patients affected by clinically probable Alzheimer's disease (AD) in order to quantify the modifications in the cortico cortical or cortico-subcortical connections. The EEGs were recorded in 10 AD patients (with mild or moderate degrees of dementia) and in 10 normal age-matched subjects, at rest and eye-closed, from 16 electrodes with linked-ears reference. Spectral parameters and coherence were calculated by a multichannel autoregressive model using 50 artifact-free epochs, 1 s duration each. Alpha coherence was significantly decreased in 6 patients, the decrease being more accentuated in the area near the electrode taken into account; a significant delta coherence increase was found in a few patients between frontal and posterior regions. The AD group showed a significant decrease of alpha band coherence, in particular in temporo-parieto-occipital areas, more evident in patients with a more severe cognitive impairment. These abnormalities could reflect two different pathophysiological changes: the alpha coherence decrease could be related to alterations in cortico-cortical connections, whereas the delta coherence increase could be related to the lack of influence of subcortical cholinergic structures on cortical electrical activity. PMID- 9743282 TI - EEG in Japanese encephalitis: a clinico-radiological correlation. AB - This study was undertaken due to the paucity of studies on electroencephalographic (EEG) changes in Japanese encephalitis (JE) and their clinical and radiological correlation. Twenty seven patients with JE were included whose age ranged between 2 and 54 years, 8 of whom were females and 10 aged 12 or less. On admission, Glasgow coma scale (GCS) ranged between 4 and 9. Seizures were present in 9 patients which were generalised tonic clonic in all except one who had partial motor seizure. Behavioural abnormalities were present in 3 patients. Three patterns of EEG were noted which included diffuse continuous delta in 21, diffuse delta with spikes in 3; and nonmodulating non responsive alpha activity ('alpha pattern' coma) in 3 patients. The background EEG activity became normal in all at 3 months although seizure activity was noted in 3 patients. MRI or/and CT scan revealed bilateral thalamic involvement in all, pons in 2, midbrain in 7, basal ganglia in 5, cerebral cortex in 4 and white matter oedema in 5 patients. Five patients died in the acute stage and 3 patients lost from follow-up. At 3 months, 7 patients had complete, 6 partial and 6 poor recovery. The EEG pattern did not correlate with the GCS and outcome. In JE, EEG reveals non-specific delta showing in acute stage and 'alpha pattern' coma may be a more common presentation than realised and does not always predict a poor outcome. PMID- 9743283 TI - Quantitative electroencephalographic evaluation of non-fatal and fatal traumatic coma. AB - Diffuse axonal injury (DAI) is an important cause of morbidity and mortality after traumatic brain injury (TBI), and its severity is therefore a major determinant of outcome. There have been suggestions that the extent of DAI may be reflected in quantitative measures of cerebral function, including the electroencephalogram (EEG) and brain-stem auditory evoked potentials (BAEPs). It has therefore been proposed that these quantitative methods of analysis may provide objective predictors of outcome following TBI. We prospectively investigated the relationship between quantitative EEG and BAEP measures and outcome, in 60 comatose patients (47 male and 13 female; age range 1-80 years, mean 36.4) after severe, closed head injury (post-resuscitation Glasgow Coma Scale (GCS) of 8). The Spearman correlation coefficients (rs) have been calculated for quantitative EEG measures (mean regional power and interhemispheric coherence) and BAEPs with patient outcome on the Glasgow Outcome and Disability Rating Scales at 6 months and 1 year. The measures most significantly correlated with outcome (P < 0.001) are over the left hemisphere, beta activity power (amplitude squared) in the fronto-central and centro-temporal regions, and alpha activity power in the centro-temporal region. We found no correlation between interhemispheric coherence (a statistical measure of cross correlation in the frequency domain) and outcome at either 6 months or 1 year post-injury. In 10 fatalities, we examined the relationship between interhemispheric EEG coherence prior to deaths and the histopathological severity of DAI, in concordant regions. The only significant correlation between DAI and interhemispheric coherence is seen in the alpha band at the temporo-occipital site (rs = -0.79, P = 0.007). Our data indicate that there is regional information in EEG power spectra over the left hemisphere, which could be used in prognostic predictions for patients in coma after severe TBI. We were unable to demonstrate a correlation between interhemispheric coherence and outcome, or any clear and consistent evidence of a relationship between interhemispheric coherence and the severity of DAI. PMID- 9743284 TI - Neuromagnetic sequelae of herpes simplex encephalitis. AB - Spontaneous cortical activity and auditory evoked responses were recorded with a whole-scalp 122-channel neuromagnetometer from 4 patients after left-hemisphere dominant herpes simplex encephalitis and associated memory disorders. Spontaneous activity of one patient contained periodic sharp waves over the left hemisphere; the background activity was attenuated. The sources of periodic sharp waves clustered close to the sources of auditory evoked fields in the temporal lobe. In controls, dominant rhythmic activity over the parieto-occipital region had spectral maximum at 10.6 +/- 0.6 Hz; in patients the dominant rhythmic activity peaked at 8.6 +/- 1.8 Hz. The suppression of the parieto-occipital activity in eyes-open versus eyes-closed condition was smaller in patients than in controls. The patients' peak spectral frequency was correlated with neuropsychological tests reflecting deficient attentional capacity. The observed changes probably reflect decreased subcortical control of the cortical electric activity. PMID- 9743285 TI - IFCN standards for digital recording of clinical EEG. International Federation of Clinical Neurophysiology. PMID- 9743286 TI - 14 and 6 Hz positive spikes preceding 3 Hz generalized spike and wave in a 15 year old patient with absence: a case report. AB - A 15 year old girl with a history of childhood absence seizures underwent a prolonged EEG recording. Twelve bursts of 3 Hz generalized spike and wave were recorded during drowsiness and light sleep, all of which were preceded by 14 and 6 Hz positive spikes with a latency of under 1 s. Nine bursts of polyspike-wave, not preceded by 14 and 6 Hz positive spikes, were recorded during deeper sleep (stage 3). The bursts of polyspike-wave were significantly longer when preceded by 14 and 6 Hz spikes than when observed in isolation. The close association between these paroxysmal events in our patient is intriguing, may be coincidental, and has never before been reported. PMID- 9743287 TI - Overexpression of the stathmin gene in a subset of human breast cancer. AB - Stathmin is a highly conserved cytosolic phosphoprotein that destabilizes microtubules. Stathmin, which has been proposed as a relay protein integrating diverse cell signalling pathways, acts in vitro as a tubulin-sequestering protein, and its activity is dramatically reduced by phosphorylation. Interestingly, stathmin expression and phosphorylation are regulated during the control of cell growth and differentiation, and there is much evidence suggesting that in vivo stathmin plays a role in the control of microtubule dynamics during mitosis. Stathmin may thus be considered as one of the key regulators of cell division. We examined 50 human primary breast tumours for stathmin mRNA and protein expression and screened for abnormalities in the chromosome region harbouring the stathmin gene. Overexpression of stathmin was found in 15 tumours (30%). At the present stage, no clear correlation emerged between stathmin expression and several prognosis markers. Interestingly, perfect matching was observed between stathmin mRNA overexpression, protein overexpression and strong staining for stathmin on paraffin-embedded tumour sections when specimens were available. Furthermore, a tentative link between loss of heterozygosity (LOH) in the 1p32-1pter region and stathmin overexpression was observed. Our results suggest that stathmin might play a role in breast carcinogenesis and that stathmin-overexpressing tumours may represent a new subtype of breast cancer. PMID- 9743289 TI - Enhanced expression of the urokinase-type plasminogen activator gene and reduced colony formation in soft agar by ectopic expression of PU.1 in HT1080 human fibrosarcoma cells. AB - To investigate the cell biological function of PU.1, a member of the Ets family of transcription factors, a vector capable of expressing the protein was transfected into HT1080 human fibrosarcoma cells. Exogenous expression of PU.1 in HT1080 cells reduced colony-forming efficiency but stimulated cell migration in soft agar, although it did not affect cell growth in adherent culture. Expression of the urokinase-type plasminogen activator (uPA) mRNA, which is known to be correlated with cell migration and invasion, was enhanced in PU.1 transfectants compared with mock transfectants. Run-on analysis demonstrated that uPA transcription was unaffected by PU.1, suggesting that this enhancement mainly occurs at a post-transcriptional level. On the other hand, treatment of HT1080 cells with the synthetic glucocorticoid dexamethasone (DEX; 10(-7) M) significantly reduced uPA gene expression at a transcriptional level. Furthermore, DEX inhibited cell migration in soft agar without affecting cell growth. These negative effects of DEX on uPA expression and cell migration were alleviated by the expression of PU.1 in HT1080 cells, whereas expression of the N ras oncogene, which is responsible for maintenance of the transformed phenotypes in HT1080 cells, was unaffected by PU.1 expression or DEX treatment in the cells. Our results suggest that expression of PU.1 can stimulate uPA gene expression at the post-transcriptional level, which may subsequently lead to activation of cell motility and/or reduced cell-cell adhesion, but reduces anchorage-independent growth of HT1080 cells. PMID- 9743288 TI - Relationship of nm23 to proteolytic factors, proliferation and motility in breast cancer tissues and cell lines. AB - Low expression of the antimetastatic gene nm23 has been associated with shorter overall survival in breast cancer. To better understand the mechanism(s) of action of this protein, we compared the levels of the nm23 protein in 152 breast cancer samples with other factors known to be involved in metastasis or related to prognosis. There was no significant relationship between either of the nm23 isoforms and cathepsin D (Cat-D), urokinase plasminogen activator (uPA), its inhibitor (PAI-1), steroid hormone receptors or ploidy status. A marginal inverse correlation was observed between per cent S-phase and nm23-H1 expression (r = 0.193, P = 0.047) and a positive correlation was observed between uPA receptor (uPAR) and both nm23-H1 (r = 0.263, P = 0.0018) and nm23-H2 (r = 0.230, P = 0.0064). The nm23-H1 gene was transfected into MDA-MB-231 human breast cancer cells and 12 clones were selected, of which two were characterized extensively. We found no significant differences in Cat-D, uPA, PAI-1 or uPAR, as a function of nm23 expression in either the MDA-MB-231 cells or the transfected clones. Compared with the parent cell line, we did observe a dose-dependent decrease in growth factor-stimulated motility and a decrease in metastatic potential in two clones with four- and eightfold elevated nm23-H1 expression, whereas the proliferative activities were similar. We conclude that the decreased metastatic potential might be related to down-regulation of growth factor-stimulated motility. PMID- 9743290 TI - Regulation of monocyte MMP-9 production by TNF-alpha and a tumour-derived soluble factor (MMPSF). AB - The matrix metalloprotease MMP-9 localizes to tumour-associated macrophages in human ovarian cancer but little is known of its regulation. Co-culture of human ovarian cancer cells (PEO-1) and a monocytic cell line (THP-1) led to production of 92-kDa proMMP-9. PEO-1-conditioned medium (CM) also stimulated THP-1 cells or isolated peripheral blood monocytes to produce proMMP-9. Expression of TIMP-1, however, remained unaffected. There was evidence that tumour necrosis factor alpha (TNF-alpha) was involved in tumour-stimulated monocytic proMMP-9 production. Antibody to TNF-alpha inhibited proMMP-9 production, and synthesis of TNF-alpha mRNA and protein preceded proMMP-9 release. In addition, the synthetic matrix metalloprotease inhibitor (MMPI) BB-2116, which blocks TNF-alpha shedding, inhibited proMMP-9 release in the co-cultures and from CM-stimulated monocytic cells. Further experiments suggested that the stimulating factor present in CM was not TNF-alpha, but acted synergistically with autocrine monocyte-derived TNF alpha to release monocytic proMMP-9. Thus, ovarian cancer cells can stimulate monocytic cells in vitro to make proMMP-9 without affecting the expression of its inhibitor TIMP-1. This induction is mediated via a soluble factor (provisionally named MMPSF) that requires synergistic action of autocrine or paracrine TNF alpha. PMID- 9743291 TI - Uptake and kinetics of 14C-labelled meta-tetrahydroxyphenylchlorin and 5 aminolaevulinic acid in the C6 rat glioma model. AB - Meta-tetrahydroxyphenylchlorin (m-THPC) and 5-aminolaevulinic acid (5-ALA) are two second-generation photosensitizers which are currently under investigation for photodynamic therapy (PDT) and photodynamic diagnosis (PDD). So far, the experience with these photosensitizers for use within brain tumours is limited. We examined the distribution and retention of 14C-labelled m-THPC and [14C]5-ALA in the rat C6 glioma brain tumour model. After intraperitoneal injection of m THPC (71,909 d.p.m. microl(-1); 0.16 mg ml(-1) m-THPC; 0.3 mg kg(-1)), the following activities were found after 36 h: brain tumour 223,664 d.p.m. g(-1), brain contralateral to the tumour side 2567 d.p.m. g(-1), liver 369,959 d.p.m. g( 1) and skin 55,197 d.p.m. g(-1); 100,000 d.p.m. corresponding to 0.22 microg of m THPC. After 7 days, the concentration of m-THPC decreased to 76,277 d.p.m. g(-1) in tumour and 635 d.p.m. g(-1) in brain. The radioactivity after intravenous administration of [14C]5-ALA (23,079 d.p.m. microl(-1); 40 mg ml(-1); 120 mg kg( 1)) increased within 15 min (59,634 d.p.m. g(-1) in tumour, 17,427 d.p.m. g(-1) in brain); after 8 h only a small amount (3653 d.p.m. g(-1) in tumour) remained. Brain adjacent to the tumour was also found to have a higher uptake of 5-ALA. This study provides basic information for the use of m-THPC and 5-ALA in brain tumours. Because of the different pharmacokinetic and toxicological profile, we recommend m-THPC for PDT and 5-ALA for PDD. Clinical trials now have to prove the superior phototoxic properties of these second-generation photosensitizers. PMID- 9743292 TI - In vitro activity of aplidine, a new marine-derived anti-cancer compound, on freshly explanted clonogenic human tumour cells and haematopoietic precursor cells. AB - Aplidine is a new marine anti-cancer depsipeptide isolated from the Mediterranean tunicate Aplidium albicans. We have evaluated its antiproliferative action against a variety of freshly explanted human tumour specimens. Concentration ranges of 0.01-1.0 microM and 0.0001-1.0 microM were used in short- and long-term exposure schedules respectively. After exposure for 1 h in 49 evaluable specimens, aplidine showed a clear concentration-dependent anti-tumour effect. At 0.05 microM, 85% of the specimens were markedly inhibited. Continuous exposure for 21-28 days in 54 tumour specimens also led to a concentration-dependent activity relationship. Fifty per cent and 100% tumour inhibitions were achieved with 0.001 microM and 0.05 microM respectively. A head to head evaluation assessing short vs continuous exposure was carried out, resulting in evidence of an activity-time of exposure relationship. Breast, melanoma and non-small-cell lung cancer appear to be sensitive to low concentrations of aplidine. In addition the evaluation of the effects of aplidine on haematopoietic cells showed a concentration-dependent toxicity. However, under continuous exposure, active concentrations induced mild bone marrow toxicity, indicating that a therapeutic window at marginally myelotoxic concentrations might exist. PMID- 9743293 TI - Induction of p53-dependent and p53-independent cellular responses by topoisomerase 1 inhibitors. AB - We have previously shown that loss of p53 function in A2780 human ovarian adenocarcinoma cells confers increased clonogenic resistance to several DNA damaging agents, but not to taxol or camptothecin. We have now extended these studies, comparing wild-type p53-expressing A2780 cells with isogenic derivatives transfected with a dominant negative mutant (143; val to ala) p53. We show that, as well as retaining equivalent clonogenic sensitivity to camptothecin, mutant p53 transfectants of A2780 cells do not acquire significantly increased resistance to the camptothecin analogues topotecan and SN-38, the active metabolite of CPT-11. Compared with vector-alone transfectants they are, however, relatively (2.2-fold) resistant to GI 147211, a further camptothecin analogue undergoing clinical trial. Treatment of A2780 with camptothecin and each analogue produces an increase, maximal at 24-48 h after drug exposure, of cells in the G2/M phase of the cell cycle and a decrease in both G1 and S-phase cells. The G2 arrest is independent of p53 function for camptothecin and the three analogues. All four compounds can induce apoptosis in A2780, which is reduced in mutant p53 transfectants, as measured using the terminal DNA transferase-mediated b-d UTP nick end labelling (TUNEL) assay. Thus, although p53-dependent apoptosis is induced by camptothecin, topotecan and SN-38 in this human ovarian carcinoma cell line, these drugs induce p53-independent death, as measured by clonogenic assay. PMID- 9743294 TI - Enhanced radiosensitivity in experimental tumours following erythropoietin treatment of chemotherapy-induced anaemia. AB - The radiosensitivity of solid tumours in anaemic rats treated with recombinant human erythropoietin (rhEPO, epoetin beta) was studied. Anaemia was induced by a single dose of carboplatin (45 mg kg(-1) i.v.), resulting in a reduction in the haemoglobin concentration by 30%. In a second group, the development of anaemia was prevented by rhEPO (1000 IU kg(-1)) administered s.c. three times per week starting 6 days before the carboplatin application. Three days after carboplatin treatment, DS-sarcomas were implanted subcutaneously onto the hind foot dorsum. Neither carboplatin nor rhEPO treatment influenced tumour growth rate. Five days after implantation, tumours were irradiated with a single non-curative dose (10 Gy), resulting in a growth delay with a subsequent regrowth of the tumours. In the rhEPO-treated group, the growth delay lasted significantly longer (9.5 days vs. 4.5 days) and the regrowth was slower (6.0 days vs. 4.1 days) compared with the anaemic group. These data suggest that the correction of chemotherapy-induced anaemia by rhEPO (epoetin beta) treatment increases tumour radiosensitivity, presumably as a result of an improved oxygen supply to tumour tissue. PMID- 9743295 TI - P-glycoprotein and metallothionein expression and resistance to chemotherapy in osteosarcoma. AB - The expression of the drug resistance (DR) mediators P-glycoprotein (P-gp) and the metallothioneins (MT) was assessed immunohistochemically in biopsy material from patients with high-grade malignant osteosarcoma (OS). No significant difference was found in survival rate between expressors of both P-gp and MT and non-expressors. Thus, it was concluded that lack of expression of these two drug resistance-related proteins does not appear to confer any advantage in terms of patient survival in osteosarcoma. PMID- 9743296 TI - Reinduction therapy with the same cytostatic regimen in patients with advanced colorectal cancer. AB - The aim of the study was to investigate the therapeutic value of reinduction therapy with the same cytostatic treatment that had been used for induction treatment in patients with metastatic colorectal cancer. A total of 71 patients, all of whom had responded or achieved stable disease lasting > or = 12 weeks after six monthly courses of first-line treatment with 5-fluorouracil + racemic leucovorin (5-FU/LV; n = 35) or 5-FU plus the l-isomer of LV (LLV; n = 34) were entered in this study. At the time of relapse, the same treatment was used for initial therapy: racemic LV or LLV was administered at 100 mg m(-2) day(-1) by i.v. bolus injection, immediately followed by 5-FU 400 mg m(-2) day(-1) given as a 2-h infusion. Chemotherapeutic drugs were given on 5 consecutive days at 4-week intervals x 6 or until there was evidence of tumour progression. Among 49 evaluable patients, reinduction therapy that was initiated after a median treatment-free interval of 5.4 months (range 3-14.5) resulted in nine partial response (PR) (18%) and 26 stable disease (SD) (53%), yielding an overall tumour control rate of 69% (95% confidence interval, 54.6-81.7%). The median time to treatment failure from reinduction was 6.4 months, and the median survival duration from reinduction was 8.9 months (20.1 months as judged from the beginning of induction therapy). The toxicity associated with retreatment was generally mild to moderate; compared with initial treatment, there was no significant difference in terms of the overall rate (P = 0.33) or severity (P = 0.19) of adverse reactions. Our data suggest that in patients with advanced colorectal cancer an interrupted treatment strategy, i.e. retreatment with the same regimen in case of relapse > or = 3 months after discontinuation of 6 months of successful treatment with 5-FU/LV or 5-FU/LLV is an acceptable therapeutic concept. PMID- 9743297 TI - Quantitative analysis of basic fibroblast growth factor and vascular endothelial growth factor in human colorectal cancer. AB - Tumour growth is angiogenesis dependent. Some authors suggest a prognostic role of microvessel count in colorectal cancer. We tested the role of basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) in the switch to the angiogenic phenotype in 35 patients with colorectal cancer at different stages of disease. We evaluated the two angiogenic factors, by enzyme-linked immunosorbent assay (ELISA), in tumour, peritumoral mucosa, pathological mesenteric and peripheral blood. We used ten endoscopic intestinal biopsies and ten peripheral blood samples from healthy subjects as control. bFGF was significantly lower in tumour tissues and in peritumoral mucosas than in healthy mucosas, whereas VEGF was up-regulated in tumours but not in peritumoral mucosa. Both angiogenic factors were greatly increased in mesenteric blood. VEGF tumour and serum levels were significantly correlated with the stage of disease. bFGF tumour and serum concentration were not correlated with the stage of disease. The high levels of bFGF in mesenteric blood suggest that this growth factor might be abnormally released from tumour tissue and peritumoral mucosa and could function as an early effector in the switch to the angiogenic phenotype. In contrast, VEGF, whose levels show a significant correlation with the stage of disease, could act in a following step, supporting tumour progression. PMID- 9743298 TI - Absence of 185delAG mutation of the BRCA1 gene and 6174delT mutation of the BRCA2 gene in Ashkenazi Jewish men with prostate cancer. AB - Epidemiological studies have demonstrated a clustering of breast and prostate cancers in some families. Moreover, there is an increase in the number of cases of prostate cancer in families with inherited mutations of the breast cancer susceptibility gene BRCA1. We assessed the role of BRCA1 and BRCA2 in prostate cancer. We tested for the BRCA1 185delAG frameshift mutation, found in 0.9% of Ashkenazi Jews, and the BRCA2 6174delT mutation, found in 1% of Ashkenazi Jews, in Ashkenazi Jewish men with prostate cancer. We studied 60 Ashkenazi men with prostate cancer. A family history was obtained by interview or a self-report questionnaire. Histological confirmation of diagnosis was obtained for all subjects. Ethnic background was confirmed for all subjects by self-report or interview. Mutations of BRCA1 and BRCA2 were detected by amplification of lymphocyte DNA from peripheral blood according to standard polymerase chain reaction (PCR) and dot blot procedures. Patients' ages ranged from 55 to 80 years (mean +/- s.d. 70 +/- 5.25). There were six men with a family history of prostate cancer; three of these had a father with prostate cancer. Five of the men had a family history of breast cancer, in a mother, a sister or an aunt. None of the men had a family history of both breast and prostate cancer. None of the 60 men carried the 185delAG BRCA1 or 6174delT BRCA2 mutations. Of 268 Ashkenazi Jewish women with sporadic breast cancer, tested in an unrelated study, 16 carried either the 185delAG mutation of BRCA1 or the 6174delT mutation of BRCA2. There was a significant difference in the incidence of the BRCA1 and BRCA2 mutations in the breast and prostate cancer cases (P = 0.05, two-tailed Fisher's exact test). The contribution of germline BRCA1 and BRCA2 mutations to prostate cancer incidence is probably small and could be limited to specific subgroups. PMID- 9743299 TI - The treatment of nephrotic syndrome caused by primary (light chain) amyloid with vincristine, doxorubicin and dexamethasone. AB - Three out of four patients with primary (light chain) amyloid nephrotic syndrome treated with vincristine, doxorubicin and dexamethasone (VAD) induction obtained a partial response and are alive in continuing remission at 4.1, 6.5 and 9.3 years. These preliminary results are of considerable interest and suggest that prospective evaluation of this regimen is warranted in patients with this condition. PMID- 9743300 TI - High-dose intensity cyclophosphamide, epidoxorubicin, vincristine and prednisone by shortened intervals and granulocyte colony-stimulating factor in non-Hodgkin's lymphoma: a phase II study. AB - Twenty patients with non-Hodgkin's lymphoma were treated with a combination of cyclophosphamide (750 mg m(-2), day 1), epidoxorubicin (60 mg m(-2), day 1), vincristine (1.4 mg m(-2), day 1) and prednisone (100 mg m(-2), days 1-5) every 14 days. Shortening of intervals was associated with the prophylactic employment of granulocyte colony-stimulating factor (G-CSF; specifically, filgrastim) administered at a dose of 300 microg subcutaneously from day 6 to day 11. The ratio between actually delivered dose intensity and planned dose intensity was 1.0 in 18 out the 20 patients. Toxicity was acceptable; response rate and survival are in the expected range. The present study demonstrated the feasibility of acceleration of chemotherapy cycles to obtain dose intensification in non-Hodgkin's lymphoma. PMID- 9743301 TI - Cost-effectiveness of recombinant human erythropoietin in the prevention of chemotherapy-induced anaemia. AB - Recombinant human erythropoietin (rHuEPO) has been advocated for the treatment of anaemia in patients submitted to cancer chemotherapy. We used decision analysis to compare the cost-effectiveness of rHuEPO supplemented with red blood cell (RBC) transfusions with conventional treatment with RBC transfusions alone. At baseline, we analysed the use of rHuEPO as secondary prophylaxis according to effectiveness estimates from a community-based oncology study. In order to reduce the probability of transfusions from 21.9% to 10.4%, and the number of RBC units per patient per month from 0.55 to 0.29, 150 units kg(-1) s.c. rHuEPO three times per week for 4 months resulted in an incremental cost of $189,652 per quality adjusted life year (QALY). In patients treated with cisplatin-containing chemotherapy, rHuEPO added $190,142 per QALY. In a hypothetical scenario of a transfusion pattern that maintained the same haemoglobin level of rHuEPO responsive patients, the marginal cost of rHuEPO was always greater than $100,000 per QALY. Results were stable with regard to variations in the probability of blood-borne infections, quality of life of responding patients and cancer-related mortality. The additional cost could be lowered to less than $100,000 per QALY by saving 4.5 RBC units over 4 months for any patient treated. In conclusion, according to current use, rHuEPO is not cost-effective in the treatment of chemotherapy-induced anaemia. More tailored utilization of the drug and better consideration of predictive response indicators may lead to an effective, blood sparing alternative. PMID- 9743302 TI - Evidence of significant apoptosis in poorly differentiated ductal carcinoma in situ of the breast. AB - Following breast-conserving surgery for ductal carcinoma in situ (DCIS), the presence of comedo necrosis reportedly predicts for higher rates of post operative recurrence. To examine the role of programmed cell death (apoptosis) in the aetiology of the cell death described as comedo necrosis, we studied 58 DCIS samples, using light microscopy, for morphological evidence of apoptotic cell death. The percentage of apoptotic cells (apoptotic index, AI) was compared between DCIS with and without evidence of 'comedo necrosis' and related to the immunohistochemical expression of the anti-apoptosis gene bcl-2, mitotic index (MI), the cellular proliferation antigen Ki67, nuclear grade and oestrogen receptor (ER) status. AI was significantly higher in DCIS samples displaying high grade comedo necrosis than in low-grade non-comedo samples: median AI = 1.60% (range 0.84-2.89%) and 0.45% (0.1-1.31%) respectively (P < 0.001). Increasing nuclear grade correlated positively with AI (P < 0.001) and negatively with bcl-2 expression (P = 0.003). Bcl-2 correlated negatively with AI (P = 0.019) and strongly with ER immunoreactivity (P < 0.001). Cellular proliferation markers (MI and Ki67 immunostaining) correlated strongly with AI and were higher in comedo lesions and tumours of high nuclear grade (P < 0.001 in all cases). Thus, apoptosis contributes significantly to the cell death described in ER-negative, high-grade DCIS in which a high proliferative rate is associated with a high apoptotic rate. It is likely that dysregulation of proliferation/apoptosis control mechanisms accounts for the more malignant features typical of ER negative comedo DCIS. PMID- 9743303 TI - Long-term survival in node-positive breast cancer treated by locoregional therapy alone. AB - To investigate the long-term survival rate of node-positive (pN+) breast cancer treated by locoregional therapy alone, we made an attempt to identify all such patients followed up for at least 15 years after treatment in a defined geographical area (city of Turku, Southwestern Finland) and time period (1945-79) using the files of the local hospitals and the Finnish Cancer Registry. The clinical and autopsy records and histological slides of 1172 women diagnosed with breast cancer in the city were reviewed. From this cohort we identified 339 women with unilateral node-positive breast cancer treated with locoregional therapy without systemic adjuvant therapy. The relative survival rate of the cohort compared with the general female population matched for age and year of follow-up was calculated. The 15- and 30-year survival rates corrected for known intercurrent deaths were 26% (95% CI, 21-31%) and 21% (16-26%) respectively, and the relative survival rates 23% and 21% respectively. None of the patients with pN2 disease survived for 15 years, whereas the 30-year corrected survival rate in pN1 disease was 24% (18-30%). Women with pT1N1M0 cancer had as high as 59% (43 75%) 15-year survival rate corrected for intercurrent deaths. A trend for improving survival was found by the decade of diagnosis. The results indicate that a considerable proportion of women with pN1 breast carcinoma treated with locoregional therapy alone become 30-year survivors and are probably cured. Adequate locoregional treatment is mandatory in the care of node-positive breast cancer. PMID- 9743304 TI - Morphometric grading of invasive ductal breast cancer. I. Thresholds for nuclear grade. AB - We analysed 170 histological samples of invasive ductal breast cancer from years 1988-91 by computerized nuclear morphometry, to find objective and quantitative thresholds for nuclear grade. Based on Kaplan-Meier curves reflecting survival and recurrence of disease and univariate analysis by Cox's regression, optimal thresholds were determined for features related to nuclear size and size variation. In our material, with mean follow-up time of 5 years 9 months, the determined thresholds for nuclear profile area (32 microm2 and 47 microm2), nuclear diameter (6.4 microm and 7.4 microm) and mean shortest nuclear axis (4.8 microm and 6.4 microm) best separated the cases with favourable, intermediate and unfavourable course of disease. In this material from the era of mammography and adjuvant therapy, the mean shortest nuclear axis was found to be a significant prognostic factor, with a risk ratio (RR) exceeded only by that of tumour size (RRs 2.9- and 3.5-fold respectively). The results suggest that morphometric grading criteria can be developed for application in Bloom-Richardson grading and in the Nottingham Prognostic Index. PMID- 9743305 TI - Chromosome alterations in breast carcinomas: frequent involvement of DNA losses including chromosomes 4q and 21q. AB - Comparative genomic hybridization was applied to map DNA gains and losses in 39 invasive ductal breast carcinomas. Frequent abnormalities included gains on chromosomal regions 1q, 8q, 11q12-13, 16p, 19, 20q and X as well as frequent losses on 1p, 5q, 6q, 9p, 11q, 13q and 16q. Furthermore, frequent losses on 4q (20 cases) and 21q (14 cases) were found for the first time in this tumour type. High copy number amplifications were observed at 8q12-24, 11q11-13 and 20q13-ter. Highly differentiated tumours were associated with gains on 1q and 11q12-13 along with losses on 1p21-22, 4q, 13q, 11q21-ter. Undifferentiated breast carcinomas were characterized by additional DNA imbalances, i.e. deletions of 5q13-23, all of chromosome 9, the centromeric part of chromosome 13 including band 13q14 and the overrepresentation of chromosome X. We speculate that these changes are associated with tumour progression of invasive ductal breast cancer. PMID- 9743306 TI - A multivariate analysis of tumour biological factors predicting response to cytotoxic treatment in advanced breast cancer. AB - The study was designed to identify factors that could predict response to chemotherapy in breast cancer. A total of 173 patients with measurable or evaluable metastatic breast cancer were enrolled in a randomized trial between November 1987 and January 1991 to receive a monthly dose of 5-fluorouracil (500 mg m(-2)), epirubicin (60 mg m(-2)) and cyclophosphamide (500 mg m(-2)) either administered in four weekly doses or in an every-4-week dose as first-line cytotoxic treatment. In 103 evaluable patients we performed a multivariate analysis of the tumour biological factors, i.e. histological grade, oestrogen receptor (ER), progesterone receptor (PR), S-phase fraction (SPF), ploidy, p53, c erbB-2, Bcl-2 and Bax expression, which showed significance in the univariate analysis according to treatment response, time to progression (TTP) or overall survival (OS). In the univariate analysis only SPF, grade and the proapoptotic protein Bax correlated with the response to cytotoxic treatment. In the multivariate analysis SPF had the strongest correlation, followed by grade and Bax. In the univariate analysis grade, PR, Bax and Bcl-2 correlated significantly with TTP, whereas in the multivariate analysis only PR showed a statistically significant correlation. In the univariate analysis PR and Bax correlated with OS and both retained its significance in the multivariate analysis. The factors that correlated significantly with the response to cytotoxic treatment in the univariate analysis, i.e. grade, SPF and Bax, seemed to predict independently the response to treatment in the multivariate analysis also. TTP and OS could be predicted partly by the same factors, although the association was quite weak. More studies and new tumour biological factors are needed to identify the group of breast cancer patients who get the most benefit from chemotherapy. PMID- 9743307 TI - MAGE, BAGE and GAGE: tumour antigen expression in benign and malignant ovarian tissue. AB - To determine if ovarian cancer patients would be suitable for MAGE-peptide vaccine-based immunotherapy, the frequency of expression of the MAGE-1-4 genes in ovarian tumours was assessed using reverse transcription polymerase chain reaction (RT-PCR) and product verification with digoxigenin-labelled oligonucleotide probes specific for each MAGE gene. In addition, the frequency of expression of more recently discovered tumour antigens (BAGE, GAGE -1, -2 and GAGE -3, -6) was established using RT-PCR and ethidium bromide staining. In this study 1/16 normal ovarian tissue specimens and 11/25 benign lesions expressed MAGE-1. In non-malignant tissue there was preferential expression of MAGE-1 in premenopausal women. A total of 15/27 malignant specimens expressed MAGE-1, including 10/14 serous cystadenocarcinomas. Expression of other tumour antigens was infrequent. The finding of MAGE-1 expression in both benign and malignant tissue questions previous assumptions regarding the role of MAGE genes in carcinogenesis. In addition, preferential MAGE-1 gene expression in non-malignant premenopausal tissue suggests that the MAGE genes may be involved in cellular proliferation as opposed to carcinogenesis or possibly that MAGE gene expression is under cyclical hormonal control. Finally, this study indicates that serous cystadenocarcinomas may be suitable tumours for MAGE-1 peptide immunotherapy. PMID- 9743308 TI - Tumour hypoxia and vascular density as predictors of metastasis in squamous cell carcinoma of the uterine cervix. AB - Some clinical studies involving several histological types of cancer have suggested that high vascular density in the primary tumour promotes metastasis. Other studies have suggested that a high incidence of metastases is associated with low oxygen tension in the primary tumour. The purpose of the study reported here was to search for correlations between incidence of metastases and oxygen tension or vascular density in the same population of patients. Thirty-eight consecutive patients with squamous cell carcinoma of the uterine cervix were included in a prospective study. Pelvic, iliac and retroperitoneal lymph node metastases were detected by magnetic resonance imaging at the time of initial diagnosis. Oxygen tension was measured polarographically using the Eppendorf pO2 Histograph 6650. Vascular density was determined by histological examination of tumour biopsies. The primary tumours of the patients with metastases (n = 19) were more poorly oxygenated than those of the patients without metastases (n = 19). Thus, the fractions of the pO2 readings resulting in values below 5 mmHg and 10 mmHg were significantly higher for the former group of patients than for the latter (P = 0.03 and 0.02 respectively). In contrast, the vascular density of the primary tumour was not significantly different for the two groups of patients. The present study suggests that a high incidence of metastases in squamous cell carcinoma of the uterine cervix is associated with poor oxygenation of the primary tumour and not with a high vascular density. PMID- 9743309 TI - Four cycles of BEP vs four cycles of VIP in patients with intermediate-prognosis metastatic testicular non-seminoma: a randomized study of the EORTC Genitourinary Tract Cancer Cooperative Group. European Organization for Research and Treatment of Cancer. AB - We investigated the efficacy and toxicity of induction chemotherapy with cisplatin and etoposide with either bleomycin or ifosfamide in patients with intermediate-prognosis testicular non-seminoma. A total of 84 eligible patients were randomized to receive four cycles of etoposide, ifosfamide, cisplatin (VIP), or four cycles of bleomycin, etoposide, cisplatin (BEP). Intermediate prognosis was defined as any of the following: lymph node metastases 5-10 cm in diameter, lung metastases more than four in number or > 3 cm, HCG 5000-50,000 IU l(-1), AFP > 1000 IU l(-1). The complete response (CR) rates to VIP and BEP were similar, 74% and 79% respectively (P = 0.62). Including the cases in whom viable cancer was completely resected with post-chemotherapy debulking surgery, the percentages of patients who achieved a no-evidence-of-disease status were 80% on VIP and 82% on BEP (P = 0.99). In addition, there were no differences in relapse rate, disease-free and overall survival after a median follow-up of 7.7 years. The 5 year progression-free survival was 85% (95% CI 74-96%) in the VIP arm and 83% (95% CI 71-96%) in the BEP arm, hazard ratio (VIP/BEP) 0.83 (95% CI 0.30-2.28). The VIP regimen was more toxic with regard to bone marrow function; the frequency of leucocytes below 2000 microl(-1) throughout four cycles was 89% on VIP and 37% on BEP (P < 0.001). Our study does not indicate that ifosfamide is superior to bleomycin in combination with cisplatin and etoposide. The sample size in this study is small as the study was prematurely discontinued when data became available from a competing study that showed no improved effectiveness of VIP compared with BEP. Taken together with these data, bleomycin should not be replaced by conventional-dose ifosfamide. PMID- 9743310 TI - Significance of plasminogen activator inhibitor 2 as a prognostic marker in primary lung cancer: association of decreased plasminogen activator inhibitor 2 with lymph node metastasis. AB - The expression of urokinase-type plasminogen activator (u-PA), u-PA receptor (u PAR) and plasminogen activator inhibitor (PAI) 1 and 2 was examined in 105 cases of primary lung cancer tissue using immunohistochemical staining and reverse transcriptase polymerase chain reaction (RT-PCR) techniques. The expression of u PA, u-PAR and PAI-1 was detected in approximately 80% of primary lung cancers, whereas detectable PAI-2 expression was observed only in half of the overall cases. We assessed the relationships between the expression pattern and clinicopathological findings and found that a diminished expression level of PAI 2 was significantly correlated with lymph node metastasis and a poor prognosis. These results indicate that PAI-2 may play a critical role in the regulation of extracellular matrix degradation during tumour cell invasion and metastasis, and the expression of PAI-2 may be useful as a marker for evaluating the prognosis of lung cancer. PMID- 9743311 TI - Presentation: Epidemiology and public health: is a new paradigm needed or a new ethic? AB - Public health militancy has been increasingly frustrated by what many perceive as the marginally fertile studies of risk factors operating at the individual level, whose causal underpinnings are often and inevitably weakened in multifactorial situations. As a remedy, leading advocates propose a refocusing of epidemiology and public health on socioeconomic, cultural, and political studies, and on broad interventions at population level. This new "paradigm" would be aided by a relaxation of evidentiary standards of causality, away from scientific criteria and more toward dialectic (rhetorical) precepts derived in a humanistic and sociologic tradition. It is countered here that such proposals would further reduce the objectivity and thus likely weaken rather than strengthen epidemiology and the justification of public health action. Instead, a realistic appraisal finds that multifactorial epidemiology raises warning signals of varying influence, and that the usefulness of epidemiology and public health could be enhanced by conceiving of methods to score the relative strength and priority of such warnings. PMID- 9743312 TI - Dissent: Back to the future in epidemiology and public health: response to Dr. Gori. PMID- 9743313 TI - More history of medical statistics. PMID- 9743314 TI - Differences in valuation of functional status components among consumers and professionals in Europe and the United States. AB - The ratings of the importance of functional status items among geriatric experts and consumers in Europe and the United States differed in many cases between experts and consumers in both countries; the differences were more frequent among the U.S. samples. The overall correlation between consumer and expert rankings was .82 for both groups. In general consumers, rated instrumental activities of daily living (IADL) items more highly, whereas the experts rated the most dysfunctional activities of daily living (ADL) items higher than did consumers. This study suggests the gap in doctor-patient communication. As function is increasingly used as a clinical outcome, agreement is needed on how to weight the components. The differences uncovered in this study suggest a need for more dialogue about what ends are truly sought by various parties. PMID- 9743315 TI - Valuing outcomes in health care: a comparison of willingness to pay and quality adjusted life-years. AB - Quality-adjusted life-years (QALYs) and willingness to pay (WTP) are two preference-based measures of health-related outcomes. In this article, we compare these two measures in eliciting individuals' preferences for health outcomes associated with shingles. To collect the necessary preference data, we administered computer-interactive interviews to a sample of 65- to 70-year-olds. We found no significant correlation between QALYs and WTP across individuals. We discuss our findings and argue that our results raise questions about whether QALYs and WTP are equivalent preference-based measures of health outcomes. PMID- 9743316 TI - Sexual dysfunction in men with lower urinary tract symptoms. AB - The conventional view that sexual function is not adversely affected by lower urinary tract symptoms (LUTS), assumed to be caused by enlargement secondary to benign prostatic hyperplasia (BPH), was investigated in this study of 423 men aged 40 years and over in a community population in the UK and 1271 urology clinic attenders aged 45 years and over in 12 countries, using the ICSmale and ICSsex questionnaires. Sexual dysfunction was found to be common: in the community, age standardized prevalences of reduced rigidity of erections were 53%, reduced ejaculation 47%, and pain on ejaculation 5%; in clinic men, age standardized prevalences of reduced rigidity of erections were 60%, reduced ejaculation 62%, and pain on ejaculation 17%. Sex lives were reported to be spoiled by LUTS in 8% of community men and 46% in the clinic. There were negative trends for age in the extent to which clinic men were bothered by these symptoms, although older men were still very concerned. Significantly raised odds ratios of sexual dysfunction were found in those with LUTS, especially storage symptoms associated with incontinence. Urinary flow rates were not associated with sexual symptoms. Sexual dysfunction is, therefore, strongly associated with LUTS, is a matter of concern to the men affected, and should be taken into account when managing patients with LUTS. PMID- 9743318 TI - Chronic symmetric symptomatic polyneuropathy in the elderly: a field screening investigation of risk factors for polyneuropathy in two Italian communities. Italian General Practitioner Study Group (IGPST). AB - A total of 4191 patients 55 years and older were screened for clinical features and risk factors of polyneuropathy by 27 general practitioners (GPs) in two areas of Italy (Varese and San Giovanni Rotondo). Polyneuropathy was diagnosed in the presence of two or more symptoms associated with bilateral impairment of at least two of the following: strength, sensation, tendon reflexes. A risk factor for polyneuropathy (associated disease or neurotoxic agent) was recorded based on its presence in the GP's records, the presence of specific treatments, or an affirmative answer to the interviewer's question. The prevalence of polyneuropathy among patients with no recognized exposure to diseases or neurotoxic agents was 1.6% (Varese 1.6%; San Giovanni Rotondo 1.8%). The corresponding values were 11.8% (Varese 11.8%; San Giovanni Rotondo 11.9%) for patients with one risk factor, and 17.3% (Varese 19.2%; San Giovanni Rotondo 13.0%) for patients with two risk factors. Combining the two populations, the prevalence of polyneuropathy was highest in diabetics (18.3%), followed by patients with a diagnosis of alcoholism (12.5%), non-alcoholic liver disease (10.9%), and tumor (7.1%). Diabetes was the commonest independent risk factor for polyneuropathy (odds ratio 11.3), followed by alcoholism (7.4), non-alcoholic liver disease (4.9), and tumor (2.6). PMID- 9743317 TI - Childhood poisoning: a population study in Trieste, Italy, 1975-1994. AB - We describe the epidemiology of 1918 cases of childhood poisoning referred to the emergency room in Trieste, Italy, from 1975 to 1994. The incidence rate of emergency room referral and subsequent hospitalization was calculated on the basis of the distribution of children resident in Trieste by calendar year. The occurrence of childhood poisoning was described according to time trends, age and gender of the child, route of exposure, symptoms at presentation to the emergency room, role of the child or others, intention, and substance involved in the poisoning. The association between presence of symptoms and characteristics of referral, host factors and substances involved was evaluated by estimating the odds ratio in multivariate models. Possible determinants of the clinical decision to treat certain cases were evaluated using logistic regression. Despite an increasing incidence of referral (from 155 per 100,000 persons per year in 1975 79 to 352 per 100,000 in 1990-94), hospital admission rates showed a two-fold decrease. Younger children (age 0-4 years) were more likely to be asymptomatic and required treatment and hospitalization less often than older children (age > or = 10 years). Trends show a decrease in pharmaceutical poisonings due probably to the introduction of child-resistant containers and an increase in domestic poisons. We also observed a steady increase in carbon monoxide inhalation and alcohol poisonings, mostly among teenagers. PMID- 9743319 TI - Skin reactions to antibacterial agents in general practice. AB - OBJECTIVES: To quantify the risk of skin reactions to antibacterial drugs under everyday circumstances in a large population with automated data from general practitioners (GP). DESIGN: A retrospective cohort study in a dynamic population. SETTING: Data came from the Integrated Primary Care Information (IPCI) database. The IPCI database consists of all data on consultations, morbidity, and prescriptions and other interventions, as registered by the GP in a source population of approximately 150,000 persons. METHODS: The study period started on April 1, 1994, and ended on September 30, 1995. All patients who were treated with an antibacterial drug were enrolled on the first day of starting treatment until the end of the study period or until the occurrence of one or more of the following diagnoses within the risk period: allergic reaction, rash, erythema, pruritus and urticaria, or a notification of a skin reaction in the free text. Subsequently, patient profiles were assessed by two authors who were blinded to exposure. The risk period was defined as the legend duration of the antibacterial drug plus 14 days to control for carry-over of drug effects and delay in patient presentation. Age, gender, and comedication were examined as potential confounders. RESULTS: In the study period 13,679 patients received 19,961 prescriptions of an antibacterial drug. It concerned 5330 men (39.0%) and 8349 (61.0%) women with a mean age of 41 and 42 years, respectively. One hundred thirty-five patients developed a skin reaction in the risk period. Rash, pruritus, urticaria, and miscellaneous skin reactions were encountered in 76 (56.3%), 18 (13.3%), 19 (14.1%), and 22 (16.3%) patients, respectively. The three most frequently reported causes of skin reactions were combinations of trimethoprim with sulfonamides (2.1% of users; incidence density [ID]: 2.1/1000 exposed days), fluoroquinolones (1.6% of users; ID: 1.5/1000 person days), and penicillins (1.1% of users; ID: 1.3/1000 person days). Compared to tetracyclines, the broad-spectrum penicillins showed an incidence density ratio (IDR) of 3.7, the combination of amoxicillin with clavulanic acid of 3.3, the fluoroquinolones of 2.8, and the combination of trimethoprim with sulphonamides of 4.4. The presence of infectious mononucleosis increased the risk of rash in amoxicillin users with a factor of 58. CONCLUSIONS: We found that the frequency of skin reactions to antibacterial drugs in general practice is around 1% and highest for the combination of trimethoprim with sulphonamides, penicillins, and fluoroquinolones. The outpatient incidence for skin reactions is probably lower than the incidence in hospitalized patients. Although this may be partly explained by negative misclassification, it is also likely that the actual incidence is lower as some hospitalized patient groups may be more prone to develop a skin reaction. PMID- 9743320 TI - Uncontrolled pearls, controlled evidence, meta-analysis and the individual patient. AB - Medicine has been dominated by uncontrolled data, often of unproven validity and insufficient to answer clinically important questions pertaining to individual patients. Controlled clinical trials, when designed and conducted rigorously, offer advantages over uncontrolled data, but they cannot be done for everything and often cater to the interests of sponsors rather than medical knowledge. With such sparse evidence, clinical research is doomed to look at main effects across populations rather than diversity of effects among individuals. By accumulating data from a large number of studies, meta-analysis provides a unique opportunity to address individual- and study-level heterogeneity. Diversity may be due to biases or may be real. Both sources must be scrutinized and meta-analysis may find a prime role in dissecting these components of diversity. Concurrent progress in basic sciences revolutionizing our predictive power for disease outcomes will heighten the importance of considering individual heterogeneity. PMID- 9743321 TI - Chemokine regulation of experimental autoimmune encephalomyelitis: temporal and spatial expression patterns govern disease pathogenesis. AB - Experimental autoimmune encephalomyelitis (EAE) is a CD4+ Th1-mediated demyelinating disease of the central nervous system that serves as a model for multiple sclerosis (MS). There are several considerations that suggest a role for chemokines in the disease process. First, chemokines are highly expressed in the central nervous system with a tight temporal relationship to disease activity. Second, in vivo neutralization studies showed a distinct role for specific chemokines in the evolution of the process. Third, the selective and differential expression of chemokines in differing models of EAE bears a close relationship to the patterns of inflammatory pathology. Fourth, the spatial distribution of chemokine expression could plausibly contribute to lesion architecture. Finally, preliminary observations in MS material suggest that chemokine expression observed in EAE may provide useful information regarding the pathogenesis of inflammation in MS. We propose that temporal and spatial expression of chemokines are crucial factors, complementing adhesion molecule up-regulation, that regulate EAE disease activity. PMID- 9743322 TI - Enhanced anti-HIV-1 activity and altered chemotactic potency of NH2-terminally processed macrophage-derived chemokine (MDC) imply an additional MDC receptor. AB - Posttranslational processing of chemokines increases (IL-8) or decreases (monocyte chemotactic protein-1) their chemotactic potency. Macrophage-derived chemokine (MDC) attracts monocytes, dendritic cells, activated lymphocytes, and NK cells and has reportedly anti-HIV-1 activity. Here we report that truncation of MDC by deletion of two NH2-terminal residues resulted in impaired binding to CC chemokine receptor (CCR)4, the only identified MDC receptor so far. Truncated MDC(3-69) failed to desensitize calcium mobilization by MDC(1-69) or thymus- and activation-regulated chemokine (TARC), another CCR4 ligand. MDC(3-69) lacked HUT 78 T cell chemotactic activity but retained its capacity to attract monocytes and to desensitize chemotaxis. Compared with MDC(1-69), MDC(3-69) had weak but enhanced antiviral activity against M- and T-tropic HIV-1 strains. Furthermore, both MDC forms failed to signal through the orphan receptors Bonzo/STRL33 and BOB/GPR15 and to desensitize RANTES and stromal cell-derived factor (SDF)-1 responses in CCR5-transfected and CXC chemokine receptor (CXCR)4-transfected cells, respectively. These findings suggest that MDC recognizes another, yet unidentified, receptor. We conclude that minimal NH2-terminal truncation of MDC differentially affects its various immunologic functions. PMID- 9743323 TI - Maternal B lymphocytes specific for paternal histocompatibility antigens are partially deleted during pregnancy. AB - Although genetically different from its mother, a mammalian fetus bearing paternal alloantigens is normally not rejected. To investigate one of the many possible mechanisms involved in this important biologic phenomenon, we analyzed the consequences of fetal alloantigen recognition on maternal B lymphocytes. We used transgenic mice expressing a unique B cell receptor with a relatively high affinity for the MHC class I molecule H-2Kk on most B lymphocytes. We provide the first evidence for an alloantigen-specific B cell deletion in the spleens and bone marrow of transgenic mothers bearing H-2Kk-positive fetuses. This highly reproducible deletion affects < or =80% of Id-bearing B cells, starts at midpregnancy, and is only observed until term. Such a specific maternal B cell deletion could contribute to the success of the fetal allograft. PMID- 9743324 TI - HindIII liposomes suppress delayed-type hypersensitivity responses in vivo and induce epidermal IL-10 in vitro. AB - Considerable evidence suggests that ultraviolet-B (UV-B) radiation suppresses certain immune responses through the induction of cyclobutane pyrimidine dimers in DNA. To determine whether induction of other forms of DNA damage in the skin mimicked the immunosuppressive effects of UV-B radiation, we produced double strand breaks in the DNA of epidermal cells with HindIII restriction endonuclease encapsulated in liposomes. Application of these liposomes, but not liposomes containing inactive HindIII or an irrelevant endonuclease, to the skin of C3H mice suppressed the induction of delayed-type hypersensitivity responses to Candida albicans and alloantigen and induced IL-10 production in the epidermis. Treatment of the Pam212 murine keratinocyte cell line with these liposomes in vitro induced immunosuppressive activity and IL-10 in culture supernatants. Unlike UV-B irradiation, however, HindIII in liposomes failed to induce suppressor T cell activity in vivo or in vitro. We conclude that double-strand breaks in DNA of epidermal cells can induce immunosuppression and up-regulate the production of immunomodulatory cytokines; however, either DNA damage alone does not account for all the immunosuppressive properties of UV-B irradiation, or cyclobutane pyrimidine dimers differ qualitatively from double-strand breaks in their biologic consequences. These studies raise the possibility that drugs causing DNA damage may induce cytokine dysregulation and immune suppression in addition to cytotoxicity. PMID- 9743325 TI - IFN-alpha is a survival factor for human myeloma cells and reduces dexamethasone induced apoptosis. AB - IFN-alpha is used as a maintenance therapy in patients with multiple myeloma, but its benefit is a matter of controversy. In vitro studies show that IFN-alpha can both stimulate and inhibit myeloma cell proliferation. We have tested the effect of IFN-alpha on the survival of myeloma cell lines and primary plasma cells. IFN alpha significantly reduced the apoptosis induced by removal of IL-6 in four IL-6 dependent myeloma cell lines. It also reduced the level of apoptosis induced by dexamethasone in these cell lines as well as in purified primary myeloma cells from seven patients. IFN-alpha promoted the survival of myeloma cells, which, following removal of IL-6, were blocked in G1 and died. However, unlike IL-6, IFN alpha-treated cells remained mainly blocked in the G1 phase of the cycle. While the effects of IL-6 are mediated through stimulation of its gp130 receptor subunit, the IFN-alpha-induced survival of myeloma cells was independent of gp130 transducer activation (as demonstrated using a neutralizing anti-gp130 Ab). However, the signal transduction cascades activated by these two cytokines share at least some common elements, since stimulation with either IFN-alpha or IL-6 resulted in STAT3 phosphorylation. These results indicate that IFN-alpha promotes the survival, but not the proliferation, of myeloma cells, preventing the apoptosis induced by removal of IL-6 or addition of dexamethasone. This survival factor activity may explain the conflicting reports on the effects of IFN-alpha on myeloma cell proliferation. PMID- 9743326 TI - Two regions in the CD80 cytoplasmic tail regulate CD80 redistribution and T cell costimulation. AB - CD28 is a major T cell costimulatory molecule, delivering signals distinct from those of the CD3/TCR complex, which regulate cytokine and cytokine receptor expression, cell proliferation, and cell viability. CD28 needs to be cross-linked to initiate signals, yet both of its ligands, CD80 and CD86, are expressed as monomers. Previously, we determined the cytoplasmic tail of CD80 is required for CD28-mediated costimulation and subcellular relocalization of CD80 in lymphocytes. In this study, we report that Reh B cell transfectants expressing CD80 with mutations in the cytoplasmic tail region either at 275-278 (RRNE- >AAAA, CD80/4A) or serine 284 (S-->A, CD80/SA) can bind ligand similar to transfectants expressing wild-type CD80, yet are unable to costimulate T cell proliferation. These mutant CD80 molecules are expressed on the surface of the Reh cells in small clusters or foci indistinguishable from those of wild-type CD80 molecules. However, mutant CD80 molecules unlike wild-type CD80 cannot be readily induced by ligand into caps. Thus, small clusters of CD80 found on APC are insufficient to initiate CD28-mediated signals, and the formation of CD80 caps appears to be a critical factor regulating the initiation of T cell costimulation. A 30-kDa phosphoprotein that associates with the cytoplasmic tail of CD80 in activated cells may play a role in CD80 redistribution and thus CD28 mediated costimulation. These results indicate two distinct regions of the CD80 cytoplasmic tail regulate its costimulatory function, and both regions are required for CD80 function. PMID- 9743327 TI - The BB1 monoclonal antibody recognizes both cell surface CD74 (MHC class II associated invariant chain) as well as B7-1 (CD80), resolving the question regarding a third CD28/CTLA-4 counterreceptor. AB - The identification of all CD28/CTLA-4 counterreceptors is critical to our understanding of this pivotal pathway of T cell activation. Clouding our understanding has been the reported discrepancies in expression and function of the B7-1 (CD80) molecule based upon the use of the BB1 vs other anti-B7-1 mAbs. To resolve this issue, we have cloned a BB1-binding molecule from the BB1+B7-1(-) NALM-6 pre-B cell line. Here, we demonstrate that this BB1-binding molecule is identical to the cell surface form of CD74 (MHC class II-associated invariant chain). CD74-transfected cells bound the BB1 mAb but not other anti-CD80 mAbs, CD28-Ig, or CTLA4Ig. Absorption and blocking experiments confirmed the reactivity of BB1 mAb with CD74. A region of weak homology was identified between CD74 and the region of B7-1 encoding the BB1 epitope. Therefore, the BB1 mAb binds to a protein distinct from B7-1, and this epitope is also present on the B7-1 protein. Many of the puzzling observations in the literature concerning the expression of human B7-1 are resolved by an understanding that BB1 staining is the summation of CD74 plus B7-1 expression. This observation requires the field to reconsider studies using BB1 mAb in the analysis of CD80 expression and function. PMID- 9743328 TI - Superclustering of B cell receptor and Fc gamma RIIB1 activates Src homology 2 containing protein tyrosine phosphatase-1. AB - Fc gamma RIIB1 (CD32) is a receptor that binds the Fc domain of Ag-complexed IgG. Coaggregation of B cell receptor (BCR) and Fc gamma RIIB1 generates a dominant negative signal that inhibits B cell activation. In Ag-specific Id-positive B cells, the co-cross-linking of BCR and Fc gamma RIIB1 by anti-Id Ab resulted in the association of both Src homology 2-containing protein tyrosine phosphatase (SHP-1) and Src homology 2-containing inositol phosphatase (SHIP) with the Fc gamma RIIB1; however, only SHIP activity was detected. "Superclustering" of the BCR and Fc gamma RIIB1 complex induced by stimulation with anti-Id Ab plus polyvalent Ag synergistically activated SHP-1. The degree of co-cross-linking between BCR and Fc gamma RIIB1 may determine the activation status of SHP-1 and SHIP. PMID- 9743329 TI - Prostaglandin E2 and dexamethasone inhibit IL-12 receptor expression and IL-12 responsiveness. AB - Regulation of the factors governing IL-12R expression and IL-12 responsiveness has been shown to be important in the generation and stability of Th1- and Th2 type responses. In this regard, cytokines have been shown to have a prominent role in regulating IL-12R expression. In this study, the role that PGE2 and dexamethasone (DXM) have in regulating IL-12R expression was evaluated. Addition of PGE2 or DXM to human PBMCs stimulated with immobilized anti-CD3 plus IL-12 inhibited the production of IFN-gamma in a dose-responsive manner. Moreover, PBMCs stimulated with immobilized anti-CD3 in the presence of PGE2 or DXM for 3 days, washed extensively, and restimulated in the presence of IL-12 still did not produce IFN-gamma. This lack of IL-12 responsiveness from cells cultured in either PGE2 or DXM was correlated with diminished surface expression of IL 12Rbeta1, IL-12Rbeta2 mRNA expression, and IL-12 binding. Finally, the PGE2- and DXM-mediated inhibition of IL-12R expression was not affected significantly by addition of neutralizing Abs against either IL-4, IL-10, or TGF-beta. By contrast, addition of dibutyryl cAMP, 8-bromoadenosine 3:5 cAMP (8-Br-cAMP), or cholera toxin substantially reduced IL-12R expression, suggesting that PGE2 may be mediating its effects through enhancement of cAMP. PMID- 9743330 TI - Fc epsilon receptor I on dendritic cells delivers IgE-bound multivalent antigens into a cathepsin S-dependent pathway of MHC class II presentation. AB - In this study, we elucidate the Fc epsilon RI-mediated Ag uptake and presentation mechanisms of dendritic cells (DC). We found that Fc epsilon RI-bound IgE, after polyvalent but not after monovalent ligation, is efficiently internalized into acidic, proteolytic compartments, degraded, and delivered into organelles containing MHC class II, HLA-DM, and lysosomal proteins. To follow the fate of the fragmented ligand, we sought to interfere with invariant chain (Ii) degradation, a process critical for peptide loading of nascent MHC class II molecules. We found DC to express cathepsin (Cat) S, a cysteine protease involved in Ii processing by B cells. Exposure of DC to a specific, active-site inhibitor of Cat S resulted in the loss of anti-Cat S immunoreactivity, led to the appearance of an N-terminal Ii remnant, and decreased the export of newly synthesized MHC class II to the DC surface. Furthermore, inactivation of Cat S as well as blockade of protein neosynthesis by cycloheximide strongly reduced IgE/Fc epsilon RI-mediated Ag presentation by DC. Thus, multimeric ligands of Fc epsilon RI, instead of being delivered into a recycling MHC class H pathway, are channeled efficiently into MIIC (MHC class II compartment)-like organelles of DC, in which Cat S-dependent Ii processing and peptide loading of newly synthesized MHC class II molecules occur. This IgE/Fc epsilon RI-dependent signaling pathway in DC may be a particularly effective route for immunization and a promising target for interfering with the early steps of allergen presentation. PMID- 9743331 TI - The role of B7-1 and B7-2 costimulation for the generation of CTL responses in vivo. AB - The role of B7-1 and B7-2 costimulatory molecules in the generation of Ag specific CD8+ CTLs is not well understood. In this paper, we analyze the role of both B7-1 and B7-2 in the generation of CTLs to nonliving, exogenous Ag and to live virus. To analyze the role of B7 costimulation in the induction of CTLs, we blocked B7-1 and/or B7-2 in vivo by injecting C57BL/6 mice with anti-B7-1 and/or anti-B7-2 mAbs; the mice were subsequently immunized with either chicken OVA that had been cross-linked to beads as a model of exogenous Ags or with wild-type and recombinant vaccinia virus expressing different forms of chicken OVA as models of viral Ags. Our results indicate that B7 costimulation is necessary in the generation of CTLs for all of these Ags. Since the B7 molecules could be costimulating CD8+ and/or CD4+ T cells in wild-type animals, we also examined the role of costimulation in the generation of CTLs to exogenous and viral Ag in MHC class II-deficient mice lacking most CD4+ T cells. In these animals, a combination of both mAbs also blocked all CTL responses, indicating that the Th cell-independent activation of CTLs is dependent upon the B7-costimulatory signals supplied to the CD8+ cell. These findings contribute to the understanding of the role of costimulation for the generation of CTLs. We also discuss the implications of these findings on the role of professional APCs in the initiation of CTL responses. PMID- 9743332 TI - The susceptibility of mice to immune-mediated neurologic disease correlates with the degree to which their lymphocytes resist the effects of brain-derived gangliosides. AB - SJL mice develop immune-mediated disorders of the central nervous system (CNS) when infected with certain neurotropic viruses or when immunized with myelin Ags. Other strains including BALB/c are more resistant to these diseases. During Sindbis virus-induced encephalitis, both mice are easily infected and elicit rapid mononuclear cell inflammation in the brain. However, only SJL mice develop immune-mediated paralysis; BALB/c mice remain asymptomatic. To understand how the same stimulus produces such divergent immunologic effects on the host, the present study investigated lymphocytes that were isolated from the brains of Sindbis virus-infected animals. Cells from the brains of SJL mice exhibited more proliferation, produced more IL-2, maintained a higher viability, and expressed less bax mRNA (a proapoptotic mediator) than did lymphocytes from the brains of BALB/c mice. Since the central nervous system is enriched in gangliosides that regulate T cell proliferation and IL-2 production in vitro, purified brain derived gangliosides were tested on peripheral lymphocytes from both strains. These lipids had less of an effect on the mitogen-induced proliferation, IL-2 production, activation-induced cell death, and up-regulation of bax mRNA in lymphocytes from SJL mice compared with those from BALB/c mice. Thus, gangliosides may inhibit various T cell effector functions and induce T cell apoptosis to a greater degree in the brains of BALB/c mice compared with the brains of SJL mice. This relative deficiency in local lymphocyte regulation may enhance the susceptibility of SJL mice to immune-mediated neurologic disease. PMID- 9743333 TI - Multiple lupus susceptibility loci map to chromosome 1 in BXSB mice. AB - BXSB mice spontaneously develop a lupus-like syndrome that is accelerated by the Yaa gene (Y-linked autoimmune accelerator). We studied the phenotype of disease in (B10 x BXSB)F1 and (BXSB x (B10 x BXSB)F1) backcross mice and genotyped 224 backcross animals to allow a microsatellite-based genome-wide linkage analysis to be conducted. In the backcross population, three intervals on chromosome 1 showed significant linkage to disease, suggesting that multiple loci contribute to the production of autoimmune disease. D1Mit5 at 32.8 cM was linked to development of nephritis (chi(2) = 15.68, p = 7.5 x 10(-5)), as was D1Mit12 at 63.1 cM (chi(2) = 20.17, p = 7.1 x 10(-6)). D1Mit403 at 100 cM was linked to anti-dsDNA Ab production (chi(2) = 17.28, p = 3.2 x 10(-5)). Suggestive linkages to antinuclear Abs and nephritis were identified on chromosome 3, to splenomegaly on chromosome 4, and to anti-ssDNA Ab production on chromosome 10. Chromosome 4 and the telomeric region of chromosome 1 have previously been linked to disease in other mouse models of systemic lupus erythematosus; however, the centromeric regions of chromosome 1 and chromosomes 3 and 10 are unique to BXSB. This implies that, though some loci may be common to a number of mouse models of lupus, different clusters of disease genes confer disease susceptibility in different strains of mice. PMID- 9743334 TI - Studies using antigen-presenting cells lacking expression of both B7-1 (CD80) and B7-2 (CD86) show distinct requirements for B7 molecules during priming versus restimulation of Th2 but not Th1 cytokine production. AB - The differentiation of CD4+ T cells into a Th1 vs Th2 phenotype profoundly influences the outcome of autoimmune and infectious diseases. B7 costimulation has been shown to affect the production of both Th1 and Th2 cytokines, depending on the system studied. There is, consequently, great interest in manipulating the B7 costimulatory signal for therapeutic purposes. To optimally manipulate this key immunoregulatory pathway, the contribution of B7 costimulation to cytokine production requires further clarification. We have compared the B7 requirement for cytokine production by naive vs previously activated T cells using DO11.10 TCR transgenic CD4+ T cells and splenic APCs from mice lacking B7 expression. Our data indicate that induction of IL-4 production and Th2 differentiation by naive T cells is highly dependent on B7 molecules, whereas IL-4 production by previously activated T cells is B7 independent. The predominant contribution of B7-mediated signals to Th1 cytokine production by both naive and primed T cells is upon IL-2 production (and expansion) rather than IFN-gamma (effector cytokine) production. Thus, our studies demonstrate that the antigenic experience of a T cell at the time of B7 blockade may determine whether blockade predominantly affects T cell expansion, differentiation, or effector cytokine production. These differential effects of B7 costimulation on IL-2 vs IFN-gamma production and on IL-4 production by naive vs primed T cells have important implications for understanding how B7:CD28/CTLA4 blockade can be effectively used to manipulate cytokine production in vivo. PMID- 9743335 TI - Granzymes D, E, F, and G are regulated through pregnancy and by IL-2 and IL-15 in granulated metrial gland cells. AB - Granulated metrial gland (GMG) cells are NK cells that proliferate and differentiate within the murine uterus during pregnancy. They have been predicted to play important roles in nurturing the embryo, normal placentation, and uterine tissue remodeling. GMG cell differentiation is manifested by the accumulation of the cytolytic mediators, perforin, granzyme A, and granzyme B, within cytoplasmic granules. The signaling mechanisms required for GMG cell differentiation are largely unknown, although recent in vitro assays have implicated IL-15 in these events. In this report, we demonstrate that granzymes D, E, F, and G (granzymes D G) are also expressed in GMG cells but at a later stage in pregnancy when compared with granzyme A expression. Whereas granzyme A is expressed in early to mid-gestation, the expression of granzymes D-G peak in mid- to late gestation. In addition, we show that the expression patterns of IL-2Rbeta and the IL-2Rgamma mRNAs overlap with that of granzyme D-G mRNAs in the pregnant uterus. Finally, we demonstrate that granzymes D-G are up-regulated by IL-2 and IL-15 in primary cultures containing GMG cells. Taken together, these results suggest that IL-2 and/or IL-15 may regulate GMG cell differentiation in vivo, and that granzymes D G may have different functions than granzyme A during pregnancy. PMID- 9743337 TI - Chimeric receptors providing both primary and costimulatory signaling in T cells from a single gene product. AB - Single chain Fv chimeric receptors, or T-bodies, are described with intracellular sequences comprising the costimulatory signaling domain of CD28 in series with the zeta-chain from the TCR complex. Using an engineered human single chain Fv derived from P67, an mAb with specificity for human CD33, and a spacer comprising an Ab hinge region with either Fcgamma or part of the CD28 extracellular region, fusion molecules were constructed to test the ability of single chain designs to mediate both primary signaling and costimulation from one extracellular binding event. Constructs with the CD28 signaling domain proximal and the zeta-chain distal to the membrane were found to express more efficiently in Jurkat than constructs with the opposite orientation and were capable of mediating up to 20 times more IL-2 production on stimulation with solid phase Ag when compared with transfectants expressing chimeric receptors with zeta-chain intracellular signaling domains only. IL-2 production was specific to Ag challenge and was completely inhibited by incubation with free Ab of the same specificity as the extracellular binding site of the construct, but not by an isotype-matched control Ab. The CD28 intracellular domain of these fusion proteins was shown to be capable of binding the p85 subunit of phosphatidylinositol 3'-kinase. These constructs represent the first of a new generation of single gene multidomain chimeric receptors capable of mediating both primary and costimulatory signaling specifically from a single extracellular recognition event. PMID- 9743336 TI - Cloning of BY55, a novel Ig superfamily member expressed on NK cells, CTL, and intestinal intraepithelial lymphocytes. AB - Expression of the BY55 protein has been shown to be tightly associated with NK and CD8+ T lymphocytes with cytolytic effector activity. To determine the function of this protein, we molecularly cloned BY55 cDNA. The cDNA sequence predicts a cysteine-rich, glycosylphosphatidylinositol-anchored protein of 181 amino acids with a single Ig-like domain weakly homologous to killer inhibitory receptors. Reduction and carboxyamidomethylation of immunoprecipitated BY55 gave a band of 27 kDa, whereas reduction alone led to an 80-kDa species, suggesting that BY55 is a tightly disulfide-linked multimer. RNA blot analysis revealed BY55 mRNAs of 1.5 and 1.6 kb whose expression was highly restricted to NK and T cells. BY55 was expressed on the CD56dim, CD16+ subset of NK cells, which have high cytolytic activity, but was not expressed and was not induced on the CD56bright, CD16-subset of NK cells, a subset with high proliferative, but low cytolytic, capacity. In human tissues, BY55 mRNA was expressed only in spleen, PBL, and small intestine (in gut lymphocytes). BY55 was expressed on all intestinal intraepithelial lymphocytes, which were predominantly CD3+TCRalpha/beta+CD4 CD8+CD11b+CD28-CD45RO+C D56-CD101+CD103+ (alphaEbeta7 integrin). In addition, BY55 was expressed on most CD8+CD28- peripheral blood T cells. These phenotypic relationships suggest that CD8+CD28+ precursor CTL may terminally differentiate into CD8+CD28-BY55+ effector CTL and that some of the peripheral blood CD8+CD28- subset may represent recirculation from mucosal epithelial immune sites. PMID- 9743338 TI - Sam68 association with p120GAP in CD4+ T cells is dependent on CD4 molecule expression. AB - p120 GTPase-activating protein (p120GAP) is a major negative regulator of p21ras activity in several cell types including T cells. Catalytic activity of this enzyme is regulated in part by its interaction with several associated tyrosine phosphorylated proteins. Sam68 was initially described as associated with p120GAP. It has been further established that Sam68 is a substrate of src kinases in mitosis and that it is not associated with p120GAP in transformed fibroblasts. We describe herein that Sam68 associates with p120GAP and PLC gamma 1 in human mature T cells and in a T cell line expressing the CD4 molecule HUT78 CD4+. This association is present in nonactivated cells and increases after anti-CD3 activation. It is dependent on CD4 expression and, in part, on the association of CD4 with p56lck, as shown by the strongly decreased association of Sam68 with p120GAP in the CD4- mutants, HUT78 CD4-, and by the reduced association of Sam68 with both p120GAP and p56lck in the HUT78 T cell line expressing a CD4 mutant unable to interact with p56lck, HUT78 C420/22. We propose that recruitment of Sam68, via CD4/p56lck, to the inner face of the plasma membrane may permit, via its docking properties, the correct association of key signaling molecules including PLC gamma 1 and p120GAP. This formation of transduction modules will enable the activation of different signaling cascades including the p21ras pathway and an array of downstream events, ultimately leading to T cell activation. PMID- 9743339 TI - Prostaglandin E2 induces the final maturation of IL-12-deficient CD1a+CD83+ dendritic cells: the levels of IL-12 are determined during the final dendritic cell maturation and are resistant to further modulation. AB - Activation of immature dendritic cells (DC) in peripheral tissues induces their migration to lymph nodes and their maturation into CD83+ DC, which are able to prime naive T cells. The inflammatory cytokines IL-1beta and TNF-alpha induce mature DC, which can secrete IL-12 and promote the development of Th0/Th1-biased cells. DC maturation factors with a Th2-promoting function have not been described. Here we show that PGE2, although it does not induce final DC maturation by itself, synergizes with IL-1beta and TNF-alpha, and allows their effectiveness at 100-fold lower concentrations. While being phenotypically identical with the DC matured in the presence of high concentrations of IL-1beta and TNF-alpha alone, DC matured in the additional presence of PGE2 show impaired IL-12 production and bias naive Th cell development toward the Th2. The ability of DC to produce IL-12 is also suppressed by IL-10, which in contrast to PGE2, inhibits their maturation. The differences in the ability to produce IL-12, established during the final DC maturation, are stable after the removal of modulatory factors. Importantly, fully mature DC become unsusceptible to PGE2 and IL-10. This indicates that the levels of IL-12 production in vivo, in mature DC interacting with Th cells within the lymph nodes, are mainly predetermined at the stage of immature DC in peripheral tissues. These data imply that the character of pathogen-induced local inflammatory reaction can "instruct" local DC to initiate Th1 or Th2-biased responses. PMID- 9743340 TI - Caspase dependence of target cell damage induced by cytotoxic lymphocytes. AB - Since the CTL secreted granule protease granzyme B can activate multiple target caspases, it has been proposed that this pathway is responsible for CTL-induced cytolysis of Fas-negative targets. However, target lysis via the granule exocytosis pathway is completely resistant to caspase inhibitors. To test the possibility that granzymes trigger a postcaspase cytoplasmic apoptotic pathway leading to lysis, we have examined the caspase dependence of several cytoplasmic changes associated with apoptotic death. Rapid prelytic phosphatidylserine externalization was induced in Jurkat target cells by both the Fas ligand (FasL)/Fas and the granule exocytosis effector pathways. This was specifically blocked by peptide ketone caspase inhibitors when induced by the former, but not by the latter, pathway. A rapid prelytic loss of target mitochondrial psi was also induced by both CTL effector pathways, and this was also specifically blocked by caspase inhibitors when induced by the FasL/Fas, but not by the granule exocytosis, pathway. Similarly, target membrane blebbing induced by CTL via the FasL/Fas, but not via the granule exocytosis, effector pathway was specifically blocked by caspase inhibitors. In contrast to the above nonnuclear damage, CTL-induced target staining by the lipid probe FM1-43 reflecting plasma membrane endocytosis was blocked by caspase inhibitors. Thus, when caspase activation is blocked, the granule exocytosis pathway triggers several parameters of target apoptotic damage in addition to lysis, suggesting that granzymes directly trigger a postcaspase cytoplasmic apoptotic death pathway. PMID- 9743341 TI - Expansion of functional NK cells in multiple tissue compartments of mice treated with Flt3-ligand: implications for anti-cancer and anti-viral therapy. AB - The generation and activity of NK cells appear to be regulated by a particular set of cytokines. We examined the in vivo effects of recombinant human Flt3 ligand (Flt3-L), a recently cloned potent hemopoietic cytokine, on NK cell development in mice. Daily i.p. administration of Flt3-L consistently induced striking increases in both the absolute number and the total cytotoxic activity of mature nonactivated NK cells within various tissues. Dose- and time-dependent increases were observed in the bone marrow (approximately 2- and approximately 11 fold, respectively), thymus (approximately 2.8- and approximately 2.0-fold), blood (approximately 11- and approximately 15-fold), spleen (approximately 10- and approximately 9-fold), and liver (approximately 15- and approximately 39 fold). In addition, IL-2 induced a rapid increase in NK activity, NK cell proliferative responses, generation of lymphokine-activated killer activity, and development of activated adherent NK cells, which were all significantly increased by Flt3-L treatment. Thus, in addition to its recently reported capacity to stimulate dendritic cell production, Flt3-L has a prominent biologic role in NK cell generation in vivo. This is probably a result of selectively induced expansion of NK cell progenitors (pro-NK cells), because Flt3-L stimulates in vitro proliferation of pro-NK cells without affecting the cytotoxicity of mature NK cells. The results also indicate that either alone or in combination with a potent activator of NK cells, such as IL-2, Flt3-L could be used to markedly augment the number and activity of NK cells, especially in the liver. Flt3-L appears to have considerable potential for therapy of both cancer and viral infection. PMID- 9743342 TI - Commitment of individual Th1-like lymphocytes to expression of IFN-gamma versus IL-4 and IL-10: selective induction of IL-10 by sequential stimulation of naive Th cells with IL-12 and IL-4. AB - Commitment of Th lymphocytes to the Th1 phenotype, as characterized by the expression of the major proinflammatory cytokine IFN-gamma, may be critically involved in the establishment of chronic inflammation and inflammatory autoimmune disease. To date, it has been shown that in IL-12-stimulated murine Th cell lines containing a major fraction of Th1 cells, Th2 cells can be induced by IL-4 until about 2 wk after initial activation, but not later. Here we analyze, based on the magnetic isolation of viable Th1 cells according to their specific expression of IFN-gamma, the cytokine commitment of individual Th1 cells. After activation of naive Th cells with Ag and IL-12 for up to 5 wk, isolated IFN-gamma-producing cells were restimulated with Ag and IL-4. Within the first 3 to 4 wk of IL-12 stimulation, some IFN-gamma+ cells stopped expression of IFN-gamma when restimulated with IL-4. However, within only 1 to 2 wk of IL-12 stimulation, few IFN-gamma+ cells could be converted to produce IL-4. Others continued to express IFN-gamma and thus were already committed to a proinflammatory, Th1-like phenotype. Surprisingly, within 3 wk of IL-12 stimulation, many of the IFN-gamma producing cells responded to IL-4 restimulation by expression of IL-10, but neither IFN-gamma nor IL-4, i.e., by conversion to a suppressive, anti inflammatory phenotype. PMID- 9743343 TI - Intracellular regulation of TRAIL-induced apoptosis in human melanoma cells. AB - The observation that TNF-related apoptosis-inducing ligand (TRAIL), a member of the TNF cytokine family, induces apoptosis in a number of different tumor cell types led us to compare the tumoricidal effects of TRAIL to those of other TNF family molecules on human melanoma cells. We found that a high proportion of the melanoma cell lines tested were killed by TRAIL, whereas all the melanoma lines were resistant to the other TNF family cytokines tested. TRAIL-induced death was characterized by caspase activation and cellular protein cleavage within minutes of TRAIL addition, and death could be completely inhibited by the caspase inhibitors Ile-Glu-Thr-Asp (IETD) and Val-Ala-Asp (VAD), indicating the presence of a TRAIL receptor signaling pathway similar to that identified for Fas and TNF receptors. Specific TRAIL receptor expression was determined by RT-PCR, and the presence of mRNA encoding the "protective" TRAIL receptors did not correspond to resistance or sensitivity to TRAIL-induced apoptosis. Addition of protein synthesis inhibitors to TRAIL-resistant melanomas rendered them sensitive to TRAIL, indicating that the presence or the absence of intracellular apoptosis inhibitors may mediate resistance or sensitivity to TRAIL-mediated apoptosis. Expression of one such inhibitor, FLICE-inhibitory protein (FLIP), was highest in the TRAIL-resistant melanomas, while being low or undetectable in the TRAIL sensitive melanomas. Furthermore, addition of actinomycin D to TRAIL-resistant melanomas resulted in decreased intracellular concentrations of FLIP, which correlated with their acquisition of TRAIL sensitivity. Collectively, our results indicate that TRAIL-induced apoptosis occurs through a caspase signaling cascade and that resistance is controlled by intracellular regulators of apoptosis. PMID- 9743344 TI - B cell tolerance to a minor, but not to a major, antigenic surface of the self antigen, cytochrome c. AB - To study B cell tolerance to the mitochondrial protein cytochrome c (CYT), the B cell response to pigeon CYT (PCC) was examined in mice transgenic for PCC. PCC was coupled to OVA to provide T cell help, since PCC-specific T cells in PCC transgenic mice are deleted in the thymus. The frequency of secondary B cells responding to the minor antigenic surface around residue 44 on PCC was decreased about 10-fold in native PCC-transgenic mice compared with that in control mice or in transgenic mice expressing an altered form of PCC that lacked the heme and had a different amino acid sequence at the N-terminus. A similar decrease has been observed in the frequency of B cells in normal mice recognizing the site around residue 44 on mouse CYT compared with the frequency of B cells recognizing the corresponding site on foreign CYT. There were no major decreases but apparently were compensatory increases in the frequencies of B cells recognizing other sites on PCC in the native PCC-transgenic mice compared with those in other mice. These results indicate that B cells in mice are only partially tolerant to self CYT. A possible basis for this partial tolerance relating to the fate of CYT in cell death is discussed. This may be the first example of the use of a transgenic system to study B cell tolerance to a homologous self Ag. PMID- 9743345 TI - Differential expression of Fas and Fas ligand in acute and chronic graft-versus host disease: up-regulation of Fas and Fas ligand requires CD8+ T cell activation and IFN-gamma production. AB - The parent-into-F1 model of acute and chronic graft-vs-host disease (GVHD) was used as an example of in vivo cell-mediated or Ab-mediated responses, respectively, and the roles of Fas and Fas ligand (FasL) were investigated. Using both flow cytometry and PCR methodologies, we found that acute GVHD mice exhibited significant up-regulation of Fas and FasL, whereas Fas/FasL up regulation in chronic GVHD mice was equal to or marginally greater than that in uninjected mice. Functional studies confirmed that Fas/FasL contributed to the anti-host CTL activity of splenocytes from acute GVHD mice, although a perforin dependent pathway was also identified. Despite the presence of FasL on both donor CD4+ and CD8+ T cells in acute GVHD mice, depletion studies demonstrated that all the in vitro anti-host CTL activity resided in the CD8+ population. Furthermore, injection of CD8-depleted B6 spleen cells into F1 mice blocked Fas/FasL up regulation and IFN-gamma production, resulting in chronic GVHD. Lastly, up regulation of Fas/FasL in acute GVHD mice could be blocked by anti-IFN-gamma mAb in vivo. Thus, in this in vivo model of alloantigen immune responsiveness, Fas/FasL up-regulation is critically dependent on Ag-specific (donor) CD8+ T cell activation and IFN-gamma production. Donor CD4+ T cell activation in the absence of CD8+ T cell activation results in an autoantibody-mediated response, no significant Fas/FasL up-regulation, impaired elimination of autoreactive B cells, and persistent humoral autoimmunity. PMID- 9743346 TI - The Th2 cytokine IL-4 is not required for the progression of antibody-dependent autoimmune myasthenia gravis. AB - Experimental autoimmune myasthenia gravis (EAMG), a disorder of the neuromuscular junction, is mediated by autoantibodies against muscle nicotinic acetylcholine receptor (AChR). The roles of IFN-gamma (Th1) and IL-4 (Th2) cytokines in the initiation and progression of this disease are not fully understood. Recently, we have demonstrated that IFN-gamma is necessary for the initiation of tAChR-induced EAMG in mice. However, the role of IL-4 in the progression of clinical EAMG remained undetermined. In this study we have addressed the contribution of IL-4 in the disease progression in IL-4(-/-) C57BL/6j mice whose IL-4 gene has been disrupted. Following immunization with Torpedo (t) AChR, the IL-4(-/-) mice readily developed signs of muscle weakness and succumbed to clinical EAMG with kinetics similar to the susceptibility of IL-4(+/+) mice. The tAChR-primed lymph node cells from IL-4(-/-) mice vigorously proliferated to tAChR and to its dominant alpha146-162 sequence associated with disease pathogenesis. However, these T cells secreted higher levels of IFN-gamma and IL-2, suggesting the development of a Th1 default pathway in these mice. Nevertheless, the IL-4 mutation had no effect on the recruitment of CD4+ Vbeta6+ T cells specific to the dominant tAChR alpha146-162 sequence in vivo. Immune sera from IL-4(-/-) mice showed a dramatic increase in mouse AChR-specific IgG2a levels followed by a concomitant decrease in IgG1 levels, but these mice did not exhibit an accelerated disease. In conclusion, we have demonstrated for the first time that IL-4 is not required either for the generation of a pathogenic anti-AChR humoral immune response or for progression of clinical EAMG in mice. PMID- 9743347 TI - IL-13 suppresses TNF-induced activation of nuclear factor-kappa B, activation protein-1, and apoptosis. AB - IL-13 is known to suppress the production of inflammatory cytokines such as TNF. Whether IL-13 also modulates the biologic effects of TNF is not known. In the present report we examined the effect of IL-13 on TNF-induced activation of nuclear transcription factors NF-kappa B and activation protein-1 (AP-1) and apoptosis. Pretreatment of cells with IL-13 blocked TNF-induced NF-kappa B activation, nuclear translocation of p65 subunit, and degradation of I kappa B alpha. IL-13 also inhibited NF-kappa B activation by LPS, okadaic acid, H2O2, and ceramide. TNF-induced NF-kappa B-dependent gene transcription was also blocked by IL-13. TNF-induced activation of another nuclear transcription factor, AP-1, was suppressed by IL-13. The activation of N-terminal c-Jun kinase and mitogen activated protein kinase kinase, implicated in the regulation of AP-1 and NF kappa B, was also down-regulated by IL-13. TNF-mediated cytotoxicity and activation of caspase-3 were abolished by IL-13. The inhibitory effects of IL-13 on TNF were sensitive to H-7, neomycin, and wortmannin, suggesting that the pathway consisting of protein kinase C, phosphatidylinositol 3-kinase, and phospholipase C must be involved in IL-13 signaling. Thus, overall, these results demonstrate that IL-13 is a potent inhibitor of TNF-mediated activation of NF kappa B, AP-1, and apoptosis, which may contribute to its previously described immunosuppressive and anti-inflammatory effects. PMID- 9743348 TI - Alpha-melanocyte-stimulating hormone inhibits the nuclear transcription factor NF kappa B activation induced by various inflammatory agents. AB - Alpha-melanocyte-stimulating hormone (alpha-MSH) is a tridecapeptide found mainly in the brain, pituitary, and circulation. It inhibits most forms of inflammation by a mechanism that is not known. As most types of inflammation require activation of NF-kappa B, we investigated the effect of alpha-MSH on the activation of this transcription factor by a wide variety of inflammatory stimuli. Electrophoretic mobility shift assay showed that alpha-MSH completely abolished TNF-mediated NF-kappa B activation in a dose- and time-dependent manner. It also suppressed NF-kappa B activation induced by LPS, okadaic acid, and ceramide. The effect was specific, as the activation of the transcription factor activating protein-1 by TNF was unaffected. Western blot analysis revealed that TNF-dependent degradation of the inhibitory subunit of NF-kappa B, I kappa B alpha, and nuclear translocation of the p65 subunit of NF-kappa B were also inhibited. This correlated with suppression of NF-kappa B-dependent reporter gene expression induced by TNF. The inhibitory effect of alpha-MSH appeared to be mediated through generation of cAMP, as inhibitors of adenylate cyclase and of protein kinase A reversed its inhibitory effect. Similarly, addition of membrane permeable dibutyryl cAMP, like alpha-MSH, suppressed TNF-induced NF-kappa B activation. Overall, our results suggest that alpha-MSH suppresses NF-kappa B activated by various inflammatory agents and that this mechanism probably contributes to its anti-inflammatory effects. PMID- 9743349 TI - Nuclear translocation of upstream stimulating factor 2 (USF2) in activated mast cells: a possible role in their survival. AB - Multiple transcription factors are activated in the cytoplasm and translocated to the nucleus where they exert positive or negative control over cellular genes. Such subcellular traffic of transcription factors usually requires the presence of a positively charged nuclear localization sequence (NLS). Upstream stimulating factor 2 (USF2) is one of the few transcription factors that contain two potential domains for nuclear localization. In addition to the conventional basic NLS, USF2 contains a highly conserved USF-specific region that is involved in its nuclear translocation. In the present work, the induction of translocation of USF2 into the mast cell nucleus was observed and found to be dependent on activation of the cells either by IL-3 or IgE-Ag. It was also observed that the prevention of the translocation of USF2 to the nucleus, using a peptide derived from the specific USF-NLS region, significantly inhibited their IL-3-mediated survival. Thus, our findings show a direct connection between mast cell surface receptor-mediated USF2 nuclear translocation and cell viability. PMID- 9743350 TI - An improved retroviral gene transfer technique demonstrates inhibition of CD4-CD8 thymocyte development by kinase-inactive ZAP-70. AB - ZAP-70 is a Syk family tyrosine kinase that plays an essential role in initiating TCR signals. Deficiency in ZAP-70 causes a defect in the development at CD4+CD8+ thymocytes due to defective TCR-mediated positive and negative selection. Using a newly devised retrovirus gene transfer and an efficient green fluorescence protein detection technique in fetal thymus organ cultures, the present study shows that forced expression in developing thymocytes of a catalytically inactive mutant of ZAP-70, but not wild-type ZAP-70, inhibits T cell development at the earlier CD4-CD8- stage. The ZAP-70 mutant blocked the generation of CD4+CD8+ thymocytes even in the absence of endogenous ZAP-70. Thus, the present results demonstrate a novel technique for gene transfer into developing T cells and suggest that ZAP-70/Syk family tyrosine kinases are involved in the signals inducing the generation of CD4+CD8+ thymocytes. PMID- 9743351 TI - Clonal analysis of a human antibody response. III. Nucleotide sequences of monoclonal IgM, IgG, and IgA to rabies virus reveal restricted V kappa gene utilization, junctional V kappa J kappa and V lambda J lambda diversity, and somatic hypermutation. AB - In previous work, we generated four IgM, five IgG1, and one IgA1 mAbs to rabies virus using B cells from four subjects vaccinated with inactivated rabies virus, a thymus-dependent (TD) mosaic Ag, and sequenced the mAb V(H)DJ(H) genes. Here, we have cloned the V kappa J kappa and V lambda J lambda genes to complete the primary structure of the Ag-binding site of these mAbs. While the anti-rabies virus mAb selection of VA genes (2e.2.2 twice, DPL11, and DPL23) reflected the representation of the V lambda genes in the human haploid genome (stochastic utilization), that of V kappa genes (O2/O12 twice, O8/O18, A3/A19, A27, and L2) did not (p = 0.0018) (nonstochastic utilization). Furthermore, the selection of both V kappa and V lambda genes by the anti-rabies virus mAbs vastly overlapped with that of 557 assorted V kappa J kappa rearrangements, that of 253 V lambda J lambda rearrangements in lambda-type gammopathies, and that of other Abs to thymus-dependent Ags, including 23 anti-HIV mAbs and 51 rheumatoid factors, but differed from that of 43 Abs to Haemophilus influenzae type b polysaccharide, a prototypic thymus-independent (TI) Ag. The anti-rabies virus mAb V kappa J kappa and V lambda J lambda segments displayed variable numbers of somatic mutations, which, in mAb58 and the virus-neutralizing mAb57, entailed a significant concentration of amino acid replacements in the complementarity-determining regions (p = 0.0028 and p = 0.0023, respectively), suggesting a selection by Ag. This Ag-dependent somatic selection process was superimposed on a somatic diversification process that occurred at the stage of B cell receptor for Ag rearrangement, and that entailed V gene 3' truncation and N nucleotide additions to yield heterogeneous CDR3s. PMID- 9743352 TI - Overexpression of BSAP/Pax-5 inhibits switching to IgA and enhances switching to IgE in the I.29 mu B cell line. AB - B cell-specific activator protein (BSAP)/Pax-5 is a paired domain DNA-binding protein expressed in the developing nervous system, testis, and in all B lineage cells, except terminally differentiated plasma cells. BSAP regulates transcription of several genes expressed in B cells and also the activity of the 3' IgH enhancer. As it has binding sites within or 5' to the switch regions of nearly all Ig heavy chain C region genes and also is known to increase transcription of the germline epsilon RNA, BSAP has been hypothesized to be involved in regulation of Ab class switch recombination. To directly examine the effects of BSAP on isotype switching, we use a tetracycline-regulated expression system to overexpress BSAP in the surface IgM+ I.29 mu B cell line, a mouse cell line that can be induced to undergo class switch recombination. We find that overexpression of BSAP inhibits switching to IgA in I.29 mu cells stimulated with LPS + TGF-beta 1 + nicotinamide, but enhances switching to IgE in cells stimulated with LPS + IL-4 + nicotinamide. Parallel to its effects on switching, overexpression of BSAP inhibits germline alpha RNA expression and the transcriptional activity of the germline alpha promoter, while enhancing activity of the germline epsilon promoter. Proliferation of I.29 mu cells is not affected in this system. The possible mechanisms and significance of the effect of BSAP on isotype switching are discussed. PMID- 9743353 TI - Sustained TCR signaling is required for mitogen-activated protein kinase activation and degranulation by cytotoxic T lymphocytes. AB - Requirements for T cell activation are not fully established. One model is that receptor occupancy and down-regulation are essential for activation, and another, not necessarily mutually exclusive, model is that sustained signals are important. Here we examine the importance of signal duration in T cell activation. First, we demonstrate that immobilized, but not soluble cross-linked, Abs to CD3 stimulate degranulation by CTL. The cross-linked Abs are not deficient in their ability to signal since they stimulate the same tyrosine phosphorylation pattern as immobilized Ab, but it is very transient relative to that stimulated by immobilized Ab. Furthermore, novel decreased migratory forms of Lck occur to a significant extent only after stimulation with immobilized Abs. A dramatic difference in the duration of signals is very evident when mitogen-activated protein kinase (MAPK) activity is examined. Immobilized anti-CD3 stimulates very high levels of MAPK activation that is still detectable 1 h after stimulation. In contrast, cross-linked Ab stimulates only transient and incomplete activation of MAPK. Taken together, these results suggest that TCR engagement and induction of tyrosine phosphorylation alone are not sufficient for T cell activation and that the duration of TCR-stimulated signals is critical to attain a functional response. PMID- 9743354 TI - HLA-DR4 (DRB1*0401) transgenic mice expressing an altered CD4-binding site: specificity and magnitude of DR4-restricted T cell response. AB - Optimum function of HLA-DR molecules in transgenic mice requires efficient interaction between the class II molecules on APCs and CD4 on T cells. Residues 110 and 139 of the second domain of class II molecules are considered to be critical for recognition of CD4. We generated an HLA-DR4beta(NT) transgene construct in which positions 110 and 139 were altered to resemble endogenous mouse H2 Abeta molecules. This construct was introduced into (B10 x SWR) embryos, and DR4beta(NT) transgenic mice were produced. The transgene was transferred into B10.RFB3 (Ebeta0 EalphaP) mice. The transgene-encoded DR4beta molecules paired with endogenous Ealpha chains to form stable DR4beta/Ealpha dimers expressed on the cell surface. The hybrid dimers showed similar Ag-binding specificity to HLA DR4 molecules and positively selected CD4+ T cells in vivo. Immunization of HLA DR4beta(NT) transgenic mice with DR4-restricted peptides induced T cell proliferation in vitro. While the purified T cells from DR4beta(NT) transgenic mice responded strongly to the HA(307-319) presented by M12C3 transfectants expressing altered DR4beta/Ealpha heterodimers, the response to the same peptides presented by transfectants expressing wild-type DR4beta/Ealpha molecules was substantially reduced. Taken together, these data confirmed in vitro studies on the importance of these residues in CD4-MHC class II interaction. The altered HLA DR4beta transgenic mice were able to overcome the species barrier and generate efficient HLA-DR4-restricted CD4-specific immune responses. Thus, residues 110 and 139 were critical for the interaction of class II with CD4 T cells during thymic selection as well as peripheral immune responses. PMID- 9743355 TI - TCR alpha beta chains associate with the plasma membrane independently of CD3 and TCR zeta chains in murine primary T cells. AB - The TCR is a multisubunit complex composed of the clonotypic alpha/beta disulfide linked heterodimer and noncovalently linked invariant CD3 gamma epsilon and CD3 delta epsilon and TCR zeta chains. Recent studies demonstrate that the surface expression of CD3 components can occur independently of the clonotypic TCR complexes in both thymocytes and splenic T cells. In this study, we report that free noncovalently associated TCR alpha beta heterodimers that exist independently of CD3 and TCR zeta chains are expressed on the cell surface of immature thymocytes and peripheral T cells, but not of T cell lines and T cell hybridomas. This suggests that the regulation of surface expression of TCR alpha beta heterodimers differs between primary T cells and T cell lines or T cell hybridomas. The isolation and biochemical characterization of surface clonotype independent CD3 complexes and free membrane-associated TCR alpha beta complexes may provide a structural basis for the quantitative difference in amount of T cell proliferation stimulated by anti-CD3 epsilon and anti-TCR beta. PMID- 9743356 TI - HIV-1 Tat inhibits human natural killer cell function by blocking L-type calcium channels. AB - Herein we show that functional phenylalkylamine-sensitive L-type calcium channels are expressed by human NK cells and are involved in the killing of tumor targets. Blocking of these channels by phenylalkylamine drugs does not affect effector/target cell binding but inhibits the release of serine esterases responsible for cytotoxicity. Interestingly, treatment of NK cells with HIV-1 Tat, which is known to affect several calcium-mediated events in immune cells, impairs their cytotoxic activity. In addition, Tat inhibits the rise in intracellular free calcium concentration upon cross-linking of the adhesion molecule CD11a, engaged during effector/target cell interaction, and the activation molecule CD16. Exogenous Tat does not influence NK-target cell binding but prevents NK cell degranulation. We propose that the molecular structure(s) on NK cells mediating the inhibitory effects HIV-1 Tat belong to L-type calcium channels, based on three lines of evidence: 1) binding of phenylalkylamine derivatives to these channels is cross-inhibited by Tat; 2) L-type calcium channels from NK cell lysates bind to Tat linked to Sepharose columns; 3) the inhibitory effect of HIV-1 Tat on NK cell function is prevented by the agonist of L-type calcium channels, Bay K 8644. Altogether, these results suggest that exogenous Tat is deeply involved in the impairment of NK cell function during HIV 1 infection. PMID- 9743358 TI - Isolation, structural characterization, and chromosomal mapping of the mouse vascular adhesion protein-1 gene and promoter. AB - Vascular adhesion protein-1 (VAP-1) is an endothelial cell adhesion molecule which mediates lymphocyte binding to endothelial cells. The cloning of a mouse VAP-1 (mVAP-1) cDNA revealed that mVAP-1 is a novel 110/220 kDa transmembrane molecule with significant identity to copper-containing amine oxidases. In this work the nucleotide sequence and primary structure of the mVAP-1 gene was determined and the promoter region was structurally characterized. The isolated approximately 14.4-kb mVAP-1 gene consists of 4 exons and 3 introns. Primer extension analysis and 5' rapid amplification of cDNA ends revealed multiple transcription initiation sites in different tissues suggesting that the mVAP-1 transcription is differently regulated in different tissues. Analysis of the sequence immediately upstream of the detected transcription initiation sites showed no canonical TATA or CCAAT elements, but putative regulatory elements were found close to the detected transcription start sites. The cloning of the mVAP-1 gene reveals the first insight into the genomic organization of murine amine oxidases and will, by targeted disruption of the gene, allow us to understand better the importance of VAP-1 in leukocyte trafficking and monoamine oxidase activity for the function of the immune system. PMID- 9743357 TI - Molecular modeling of an anti-DNA autoantibody (V-88) and mapping of its V region epitopes recognized by heterologous and autoimmune antibodies. AB - Anti-DNA autoantibodies are a characteristic feature of human systemic lupus erythematosus (SLE) and lupus diseases in the mouse. V-88 is an IgG1/kappa ssDNA binding Ab, derived from a lupus mouse, that bears a cross-species, cross reactive Id (CRI) that has been implicated in the pathogenesis of both human and murine disease. A linear epitope map of V-88 has been determined with anti idiotypic antisera obtained from rabbits, and candidate sequences for the idiotopes of the CRI have been proposed. We now report the modeling of the three dimensional structure of the V regions of Ab V-88, to map the location of these idiotopes. The V region framework structure was derived from those of crystallographically determined Ab structures, and the complementarity determining region (CDR) structures were based upon the set of canonical structures adopted by these loop regions in Abs of known structure. One of the idiotopes is an extensive, highly accessible epitope consisting of framework regions spatially adjacent to CDR2 in the heavy chain. Epitopes recognized by an anti-idiotypic rabbit antiserum were compared with those recognized by autoimmune sera from SLE-prone mice, and common features were identified. By analogy with the crystal structure of an anti-DNA Ab BV04-01 complexed with a trinucleotide, the modeled structure also suggests a mode of binding of ssDNA to V-88. The location of the candidate CRI, although within the framework region of VH, is such that it could influence Ag specificity. PMID- 9743359 TI - The inability of the nonobese diabetic class II molecule to form stable peptide complexes does not reflect a failure to interact productively with DM. AB - Sequence variability in MHC class II molecules plays a major role in genetically determined susceptibility to insulin-dependent diabetes mellitus (IDDM). It is not yet clear whether MHC class II polymorphism allows selective binding of diabetogenic peptides or regulates some key intracellular events associated with class II-restricted Ag presentation. In this study, we have employed gene transfer techniques to analyze the intracellular events that control peptide acquisition by the unique class II molecule expressed by nonobese diabetic mice (I-Ag7). This structurally unique class II molecule fails to demonstrate stable binding to antigenic peptides and fails to undergo the conformational change associated with stable peptide binding to class II molecules. The experiments reported here demonstrate that I-Ag7 can productively associate with two protein cofactors important in class II-restricted Ag presentation, invariant chain (Ii) and DM. DM participates in the removal of the Ii-derived class II-associated Ii chain peptide and the p12 degradation product from the I-Ag7 molecule. In addition, I-Ag7 undergoes a conformational change when DM is expressed within the APC. Finally, DM can mediate accumulation of peptide/class II complexes on the surface of APCs. Collectively, our experiments indicate that the failure of the I Ag7 molecule to stably bind peptide cannot be attributed to a failure to interact with the DM or Ii glycoproteins. PMID- 9743360 TI - Limited diversity of human scFv fragments isolated by panning a synthetic phage display scFv library with cultured human melanoma cells. AB - To broaden the specificity of the Abs recognizing human melanoma-associated Ags (MAAs), we have isolated human single-chain fragment of the V region (scFv) fragments by panning the synthetic phage Ab library (#1) with the human melanoma cell lines S5 and SK-MEL-28. All of the isolated scFv fragments reacted with the mouse mAb defined high molecular weight melanoma-associated Ag (HMW-MAA). scFv #70 immunoprecipitates the two characteristic subunits of HMW-MAA, while scFv #28 only immunoprecipitates its large subunit. These results challenge the current view regarding the structure of HMW-MAA and indicate that it consists of two independent subunits. The human scFv fragments share some similarities with the mouse anti-HMW-MAA mAb. Like mAb 149.53 and 225.28, scFv #28 reacts with rat B49 neural cells that express a homologue of HMW-MAA. scFv #70 reacts with a determinant that is spatially close to the one identified by mAbs 149.53, VT68.2, and VT86. Besides suggesting similarities in the recognition of human melanoma cells by the mouse and human Ab repertoire, these results indicate that the Abs isolated from synthetic Ab libraries resemble those that are found in natural Ab repertoires. The restricted diversity of the scFv fragments that were isolated by panning synthetic Ab libraries with different melanoma cell lines suggests that certain Ags, like HMW-MAA, are immunodominant in vitro. This phenomenon, which parallels the in vivo immunodominance of certain Ags, implies that the antigenic profile of the cells used for panning determines the specificity of the preponderant population of isolated Abs. PMID- 9743361 TI - Molecular cloning of two isoforms of the guinea pig C3a anaphylatoxin receptor: alternative splicing in the large extracellular loop. AB - The anaphylatoxin C3a is released from C3 during complement activation. C3a is a potent spasmogen and has recently been described as an eosinophil and mast cell chemotactic factor that mediates a number of inflammatory reactions. Previously, we demonstrated the presence of a specific C3a receptor (C3aR) on guinea pig platelets. We report here the isolation of cDNA clones encoding for two isoforms of guinea pig C3aR (gpC3aR). Hydropathy analysis of the deduced amino acid sequence of both gpC3aR clones indicated seven transmembrane domains with a large extracellular (EC) loop between the fourth and fifth transmembrane domains, which is a known characteristic of the human C3aR. Northern blot analysis revealed that the gpC3aR was abundantly expressed on macrophages and in the spleen. A comparison of the deduced amino acid sequence of the larger gpC3aR (gpC3aR-L) with the recently cloned human C3aR indicated a 59.5% identity. The deduced amino acid sequence of the second, smaller cDNA clone was identical with gpC3aR-L, except that it lacked 35 amino acids in the large EC loop. Our evidence indicates that alternative splicing occurred in the large EC loop that accounts for these two isoforms. L cells separately expressing one of these two isoforms of the gpC3aR showed similar high-affinity C3a binding. An RT-PCR analysis documented that both forms of the C3aR were expressed in a variety of guinea pig tissues. The cloning and expression of these two natural forms of gpC3aR cDNA indicated that the deletion of the 35-residue portion of the large EC loop of gpC3aR-L did not alter C3a binding. PMID- 9743362 TI - The MHC class I-restricted immune response to HIV-gag in BALB/c mice selects a single epitope that does not have a predictable MHC-binding motif and binds to Kd through interactions between a glutamine at P3 and pocket D. AB - Using a strain of Listeria monocytogenes that stably expresses and secretes HIV gag to deliver this Ag to the MHC class I pathway of Ag processing, we have identified the immunodominant CTL epitope to gag in the BALB/c mouse and shown that it is Kd restricted. The specific motif for the peptides that bind the MHC class I molecule H-2 Kd is believed to be a nonamer with residues tyrosine or phenylalanine in the second amino acid position and leucine or isoleucine in the carboxyl-terminal or ninth amino acid position as dominant anchoring positions. Surprisingly, the identified gag peptide, AMQMLKETI, does not contain an anchoring aromatic residue in position two although competition assays with other Kd-restricted epitopes indicated that it binds to Kd with comparable affinity. Using a theoretical molecular dynamics approach to probe the stability of peptide binding to MHC class I molecules, we show that the absence of an appropriate anchor residue at P2 in AMQMLKETI is compensated by favorable interactions of the glutamine at P3 with pocket D of Kd. These findings were verified experimentally, demonstrating the predictive power of this theoretical approach in analyzing MHC class I/peptide interactions. These studies also indicate that CTL epitope prediction that relies on dominant peptide motifs may not always identify the correct epitope. PMID- 9743363 TI - Inducible nitric oxide synthase is not required for long-term vaccine-based immunity against Toxoplasma gondii. AB - Induction of reactive nitrogen intermediates by IFN-gamma is presumed an important mechanism of host resistance against acute and chronic infection with Toxoplasma gondii. Although nitric oxide (NO) has been shown to be important in the control of parasite replication in vivo, the role of this molecule in vaccine based immunity against T. gondii is unknown. Mice with a targeted disruption of inducible NO synthase (iNOS) were immunized with an avirulent temperature sensitive strain of this parasite (ts-4). Both the parental C57BL/6 and the iNOS( /-) mice survived infection with the ts-4 mutant. Oral challenge of the vaccinated mice with a lethal dose of cysts containing bradyzoites resulted in reduced parasite burden and increased survival compared with nonvaccinated control mice. Host immunity in the iNOS(-/-) mice, similar to that observed in the parental strain, appears dependent upon both IFN-gamma and CD8+ T cells. These findings suggest that although vaccine-based long-term immunity against T. gondii is dependent upon the induction of IFN-gamma, it does not rely upon the anti-microbial effect of NO. PMID- 9743364 TI - Construction of a lipopolysaccharide reporter cell line and its use in identifying mutants defective in endotoxin, but not TNF-alpha, signal transduction. AB - Gram-negative bacterial LPS is a potent activator of inflammatory responses. The binding of LPS to CD14 initiates signal transduction; however, the molecular processes immediately following this event remain unclear. We engineered an LPS inducible fibroblast reporter cell line to facilitate the use of molecular genetic techniques to study the LPS signaling pathway. A plasmid containing the human Tac Ag cDNA under transcriptional control of the human E selectin promoter was cotransfected into Chinese hamster ovary (CHO)-K1 cells together with a CD14 expression plasmid. A cell line was obtained, 3E10, which upregulated expression of Tac following stimulation with LPS. Pools of mutagenized cells were exposed to LPS and then labeled with anti-Tac mAb. Cells that failed to up-regulate Tac expression were enriched by flow cytometry. Thirty clonal mutant cell lines were identified that continued to express CD14 and bind LPS, but failed to express Tac or translocate nuclear factor-kappaB (NF-kappaB) following LPS exposure. TNF alpha-treated mutant cells continued to express Tac and translocate NF-kappaB. An analysis of LPS-induced NF-kappaB activity in heterokaryons derived from polyethylene glycol-fused cell lines indicated that recessive mutations in genes encoding components of the LPS signaling pathway accounted for the signaling defects. To date, two complementation groups have been identified from 11 cell lines analyzed. These data demonstrate that the TNF-alpha signaling pathway diverges from the LPS pathway early in the signal-transduction cascade despite similarities in LPS- and TNF-alpha-induced responses. Identification of the genes affected in these mutant reporter cells should identify heretofore-elusive components of the LPS signaling cascade. PMID- 9743365 TI - Two distinct phospholipases C of Listeria monocytogenes induce ceramide generation, nuclear factor-kappa B activation, and E-selectin expression in human endothelial cells. AB - Infection of endothelial cells by Listeria monocytogenes is an essential step in the pathogenesis of listeriosis. We recently reported that L. monocytogenes induces up-regulation of E-selectin and other endothelial adhesion molecules and subsequent polymorphonuclear leukocyte (PMN) adhesion into cultured human endothelial cells. In the present study, we characterized the mechanisms of enhanced E-selectin expression using L. monocytogenes wild type (EGD), the isogenic in-frame deletion mutants for phosphatidylcholine (PC)- and phosphatidylinositol (PI)-specific phospholipases EGD delta plcA and EGD delta plcB, as well as the nonvirulent control strain Listeria innocua. Infection of endothelial cells with EGD delta plcA or EGD delta plcB for 6 h induced, as compared with EGD wild type, intermediate levels of E-selectin mRNA and protein as well as PMN rolling and adhesion at a shear rate of 1 dyne/cm2, indicating that both bacterial phospholipases are required for a maximal effect. Similarly, ceramide content and NF-kappa B activity were increased in L. monocytogenes exposed endothelial cells, but only to intermediate levels for PC- or PI phospholipase C (PLC)-deficient listerial mutants. Phospholipase effects could be mimicked by exogenously added ceramides or bacterial sphingomyelinase. The data presented indicate that PI-PLC and PC-PLC are important virulence factors for L. monocytogenes infections that induce accumulation of ceramides that in turn may act as second messengers to control host cell signal-transduction pathways leading to persistent NF-kappa B activation, increased E-selectin expression, and enhanced PMN rolling/adhesion. The ability of L. monocytogenes to stimulate PMN adhesion to endothelial cells may be an important mechanism in the pathogenesis of severe listeriosis. PMID- 9743366 TI - Prostaglandin E2 protects against liver injury after Escherichia coli infection but hampers the resolution of the infection in mice. AB - cAMP-increasing agents such as prostaglandin E2 (PGE2) are known to protect against LPS-induced liver injury by downregulating the production of inflammatory cytokines such as TNF-alpha. However, the effects of such reagents on host defense against bacterial infection remain unknown. We show here that in vivo administration of PGE2 significantly protected mice against liver injury after Escherichia coli infection but hampered the resolution of the infection. PGE2 significantly suppressed circulating TNF-alpha and IL-12 levels but increased the IL-10 production after E. coli challenge. PGE2 inhibited the emergence of gammadelta T cells in the peritoneal cavity, which are important for host defense against E. coli, and deteriorated bacterial exclusion in the peritoneal cavity after E. coli challenge. These results suggested that PGE2 affects host defense mechanisms against E. coli infection through modulation of cytokine production and gammadelta T cell accumulation. PMID- 9743367 TI - Murine IgG1 complexes trigger immune effector functions predominantly via Fc gamma RIII (CD16). AB - Previously, we have demonstrated that phagocytosis of IgG1-coated particles by macrophages in vitro is impaired by deletion of Fc gamma RIII in mice, suggesting that IgG1 may interact preferentially with Fc gamma RIII. In the present study, the biologic relevance of this observation was addressed by triggering various effector functions of the immune system in Fc gamma RIII(-/-) mice, using panels of mAbs of different IgG subclasses. Both binding and phagocytosis of IgG1-coated sheep or human erythrocytes by Fc gamma RIII(-/-) macrophages in vitro were strongly impaired, indicating that the impaired ingestion of complexed IgG1 by Fc gamma RIII(-/-) macrophages is due to a defect in binding. An in vivo consequence of the defective phagocytosis was observed by resistance of Fc gamma RIII deficient mice to experimental autoimmune hemolytic anemia, as shown by a lack of IgG1-mediated erythrophagocytosis in vivo by liver macrophages. Furthermore, trapping of soluble IgG1-containing immune complexes by follicular dendritic cells in mesenteric lymph nodes from Fc gamma RIII(-/-) mice was abolished. Whole blood from Fc gamma RIII(-/-) mice was unable to induce lysis of tumor cells in the presence of IgG1 antitumor Abs. Finally, IgG1 mAbs proved unable to mount a passive cutaneous anaphylaxis in Fc gamma RIII(-/-) mice. Together, these results demonstrate that IgG1 complexes, either in particulate or in soluble form, trigger in vitro and in vivo immune effector functions in mice predominantly via Fc gamma RIII. PMID- 9743368 TI - Gene-modified tumor vaccine with therapeutic potential shifts tumor-specific T cell response from a type 2 to a type 1 cytokine profile. AB - Vaccination with a poorly immunogenic/nonimmunogenic tumor fails to protect the host from a subsequent challenge with the same tumor. The mechanisms underlying the failure of these tumors to sensitize therapeutic T cells are not clearly understood, but the inability of host T cells to recognize tumor has been implicated. In this study, vaccination with the poorly immunogenic B16BL6-D5 (D5 H-2b) tumor fails to generate therapeutic T cells from the tumor vaccine-draining lymph nodes (TVDLN) in our adoptive immunotherapy model. However, if vaccination is performed with an allogeneic MHC class I gene (H-2 Kd)-modified tumor, the T cells obtained from the TVDLN are therapeutic after activation with anti-CD3 and IL-2. Lymph nodes (LN) draining both D5 and D5-Kd tumor vaccines contained increased numbers of cells with reduced expression of L-selectin (L-selectin(low/ )) compared with naive LN. This implies that vaccination led to sensitization of T cells even in LN draining the unmodified D5 tumor. L-selectin(low/-) cells from D5-Kd, but not D5, TVDLN were therapeutic in our animal model. No antitumor activity was seen in the high level L-selectin T cells. L-selectin(low/-) T cells exhibited tumor-specific cytokine release that was type 2 (IL-4, IL-10) following vaccination with native D5 and type 1 (IFN-gamma) following vaccination with gene modified D5-Kd. Our data suggest that the failure of unmodified D5 to generate therapeutic T cells is not due to an inability to recognize tumor Ags, but, rather, to the induction of an immune response that is ineffective in mediating tumor regression, i.e., immune deviation. PMID- 9743369 TI - Activation of cutaneous dendritic cells by CpG-containing oligodeoxynucleotides: a role for dendritic cells in the augmentation of Th1 responses by immunostimulatory DNA. AB - Genetic vaccination depends at least in part on the adjuvant properties of plasmids, properties that have been ascribed to unmethylated CpG dinucleotides in bacterial DNA. Because dendritic cells (DC) participate in the T cell priming that occurs during genetic vaccination, we reasoned that CpG-containing DNA might activate DC. Thus, we assessed the effects of CpG oligodeoxynucleotides (CpG ODN) on Langerhans cell (LC)-like murine fetal skin-derived DC (FSDDC) in vitro and on LC in vivo. Treatment with CpG ODN as well as LPS induced FSDDC maturation, manifested by decreased E-cadherin-mediated adhesion, up-regulation of MHC class II and costimulator molecule expression, and acquisition of enhanced accessory cell activity. In contrast to LPS, CpG ODN stimulated FSDDC to produce large amounts of IL-12 but only small amounts of IL-6 and TNF-alpha. Injection of CpG ODN into murine dermis also led to enhanced expression of MHC class II and CD86 Ag by LC in overlying epidermis and intracytoplasmic IL-12 accumulation in a subpopulation of activated LC. We conclude that immunostimulatory CpG ODN stimulate DC in vitro and in vivo. Bacterial DNA-based vaccines may preferentially elicit Th1-predominant immune responses because they activate and mobilize DC and induce them to produce large amounts of IL-12. PMID- 9743370 TI - Human T cell leukemia virus-I (HTLV-I) Tax-mediated apoptosis in activated T cells requires an enhanced intracellular prooxidant state. AB - We have shown that an estradiol-dependent activation of human T cell leukemia virus-I Tax leads to the inhibition of cell proliferation and to the induction of apoptosis. The present study demonstrates that a hormone-dependent activation of Tax promotes an enhanced prooxidant state in stably transfected Jurkat cells as measured by changes in the intracellular levels of glutathione and H2O2; these changes are followed by apoptotic cell death. Additional stimulation of the CD3/TCR pathway enhances the oxidative and apoptotic effects. Both Tax-mediated apoptosis and oxidative stress can be potently suppressed by antioxidants, as is seen with the administration of recombinant thioredoxin (adult T cell leukemia derived factor) or pyrrolidine dithiocarbamate. Hormone-induced Tax activation induces a long-lasting activation of NF-kappaB, which is a major target of reactive oxygen intermediates. The long-term exposure of Jurkat cells to hormone eventually results in a selection of cell clones that have lost Tax activity. A subsequent transfection of these apparently "nonresponsive" clones allows the recovery of Tax responses in these cells. Our observations indicate that changes in the intracellular redox status may be a determining factor in Tax-mediated DNA damage, apoptosis, and selection against the long-term expression of Tax function. PMID- 9743371 TI - Treatment of Paracoccidioides brasiliensis-infected mice with a nitric oxide inhibitor prevents the failure of cell-mediated immune response. AB - The activation of the nitric oxide (NO) production system and its involvement in the control of the lung fungal burden and in immunosuppression mechanisms were studied during the course of Paracoccidioides brasiliensis-infected mice. Mice that had been infected with the fungus were treated daily with a specific inhibitor of NO synthesis, N omega-nitro-L-arginine, or with buffered saline (control); NO production was assessed on the basis of spontaneous NO2- production by bronchoalveolar and peritoneal macrophages (Mphi) and of serum NO3- levels. The infection coursed with an elevation of NO3- levels. The Mphi produced NO2- and released TNF-alpha only after stimulation with LPS. In addition, the immunoproliferative responses of spleen cells that had been stimulated with the fungus Ag or with Con A were depressed. An examination of the lungs of infected animals showed a progressive increase in the size of the lesions. Treatment of the animals, which resulted in an inhibition of NO2- production by Mphi and a reduction of serum NO3- levels, caused the spontaneous release of TNF-alpha from infected animals and prevented the failure of the lymphoproliferative capacity of spleen cells. Furthermore, the treatment resulted in less pulmonary damage despite the fact that the lung fungal burden increased. It was also demonstrated that the NO donors S-nitroso-acetyl penicillamine and 3-morpholino-sydnonimine hydrochloride were able to inhibit the growth of P. brasiliensis in vitro. These results suggest that although NO is important for the killing of the fungi, the activation of NO production in P. brasiliensis infection contributes to the occurrence of the immunosuppression observed during the course of the infection. PMID- 9743372 TI - Triple role of platelet-activating factor in eosinophil migration across monolayers of lung epithelial cells: eosinophil chemoattractant and priming agent and epithelial cell activator. AB - Infiltration of eosinophils into the lung lumen is a hallmark of allergic asthmatic inflammation. To reach the lung lumen, eosinophils must migrate across the vascular endothelium, through the interstitial matrix, and across the lung epithelium. The regulation of this process is obscure. In this study, we investigated the migration of human eosinophils across confluent monolayers of either human lung H292 epithelial cells or primary human bronchial epithelial cells. Established eosinophil chemoattractants (IL-8, RANTES, platelet-activating factor (PAF), leukotriene B4, and complement fragment 5a (C5a)) or activation of the epithelial cells with IL-1beta induced little eosinophil transmigration (<7% in 2 h). In contrast, addition of PAF in combination with C5a induced extensive (>20%) transepithelial migration of unprimed and IL-5-primed eosinophils. Eosinophil migration assessed in a Boyden chamber assay, i.e., without an epithelial monolayer, was only slightly increased upon addition of PAF and C5a. Preincubation of eosinophils with the PAF receptor antagonist WEB 2086 only inhibited migration of unprimed eosinophils toward PAF and C5a, whereas preincubation of epithelial cells with WEB 2086 abolished migration of both IL-5 primed and unprimed eosinophils. This latter result indicated the presence of PAF receptors on epithelial cells. Indeed, addition of PAF to epithelial cells induced an increase in cytosolic free Ca2+, which was blocked by the PAF receptor antagonists WEB 2086 and TCV-309. Our results show that PAF induces permissive changes in epithelial cells, and that PAF acts as a chemoattractant and priming agent for the eosinophils. PMID- 9743373 TI - Differential regulation of monocyte matrix metalloproteinase and TIMP-1 production by TNF-alpha, granulocyte-macrophage CSF, and IL-1 beta through prostaglandin-dependent and -independent mechanisms. AB - Matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs (TIMPs) produced by monocytes are believed to be involved in the migration of these cells through the basement membrane and the ensuing destruction of connective tissue in chronic inflammatory lesions. Because monocytes encounter a variety of cytokines at these sites, we examined the effect of cytokines either alone or in combination on the production of monocyte MMPs and TIMP-1. TNF-alpha, granulocyte-macrophage-CSF (GM CSF), or IL-1 beta when added individually enhanced the endogenous levels of 92 kDa gelatinase (MMP-9) and TIMP-1 but failed to induce interstitial collagenase (MMP-1). However, GM-CSF, when added with either TNF-alpha or IL-1 beta, induced MMP-1 and synergistically enhanced MMP-9 and TIMP-1. Th2 cytokines, such as IL-4, inhibited the induction of MMPs and TIMP-1 by TNF-alpha, GM-CSF, and IL-1. Cytokine stimulation of MMP-1 was due, at least in part, to an increase in the release of arachidonic acid and PG E2 (PGE2), because inhibition of MMP-1 by indomethacin could be reversed by exogenous PGE2. In contrast to MMP-1, cytokine stimulation of MMP-9 and TIMP-1 was unaffected by indomethacin. The PGE2 independent induction of monocyte MMP-9 and TIMP-1 by these cytokines differed from stimulation of MMP-9 and TIMP-1 by LPS, which is in large part PG-dependent. In addition, LPS stimulated higher levels of MMP-1 whereas cytokines induced higher levels of MMP-9 and TIMP-1. This is the first demonstration that monocyte MMP-1 can be induced by cytokines and that MMP-1, MMP-9, and TIMP-1 are differentially regulated by cytokines through PG-dependent and -independent mechanisms. PMID- 9743374 TI - Evidence for the involvement of Fas ligand and perforin in the induction of vascular leak syndrome. AB - Endothelial cell injury resulting in vascular leak syndrome (VLS) is one of the most widely noted phenomenons in a variety of clinical diseases. In the current study we used IL-2-induced VLS as a model to investigate the role of cytolytic lymphocytes in the cytotoxicity of endothelial cells. Administration of IL-2 (75,000 U/mouse, three times a day for 3 days) into BL/6 wild-type mice triggered significant VLS in the lungs, liver, and spleen. Interestingly, perforin-knockout (KO) mice exhibited a marked decrease in IL-2-induced VLS in all three organs tested. Also, Fas ligand-defective (gld) mice and Fas-deficient (lpr) mice exhibited decreased VLS in the liver and spleen, but not in the lungs. The decreased VLS seen in perforin-KO, gld, and lpr mice was not due to any defect in lymphocyte migration or homing to various organs because histopathologic studies in these mice demonstrated significant and often greater perivascular infiltration of lymphocytes compared with the IL-2-treated wild-type mice. Ultrastructural studies of the lungs demonstrated significant damage to the endothelial cells in IL-2-treated wild-type mice and decreased damage in perforin KO mice. IL-2 administration caused up-regulation of CD44 in all strains of mice tested and triggered increased LAK activity against an endothelial cell line in wild-type and gld mice, but not in perforin-KO mice. The current study demonstrates for the first time that perforin and Fas ligand may actively participate in endothelial cell injury and induction of VLS in a variety of organs. PMID- 9743375 TI - SFA-1/PETA-3 (CD151), a member of the transmembrane 4 superfamily, associates preferentially with alpha 5 beta 1 integrin and regulates adhesion of human T cell leukemia virus type 1-infected T cells to fibronectin. AB - In this study we have analyzed the adhesion molecules associated with and the biologic function of SFA-1/PETA-3 (CD151) in human T cell leukemia virus type 1 (HTLV-1)-infected T cells and in freshly isolated adult T cell leukemia (ATL) cells using an anti-CD151 mAb. The anti-CD151 mAb coprecipitated alpha 5 beta 1 integrin from HTLV-1-infected T cells. Conversely, an anti-alpha 5 integrin mAb coprecipitated CD151. The anti-CD151 mAb inhibited the adhesion of HTLV-1 infected T cells to fibronectin but did not have any effect on their adhesion to laminin, collagen type I, or collagen type IV. Moreover, antisense CD151 oligonucleotide-treated HTLV-1-infected T cells showed significant inhibition of adhesion to fibronectin. These findings showed that the CD151 molecule was associated with the alpha 5 beta 1 integrin molecule and that it enhanced alpha 5 beta 1 integrin-mediated adhesion to fibronectin. In addition, the expression levels of CD151, alpha 4 beta 1 integrin, and alpha 5 beta 1 integrin on ATL cells from lymph nodes of lymphoma-type ATL patients were significantly higher than those on circulating ATL cells from leukemia-type ATL patients. This suggests that the increased expression of these integrins may contribute to lymphoma formation through the adhesion of ATL cells to the extracellular matrix and dendritic cells, rather than contributing to transmigration. PMID- 9743376 TI - EBI1/CCR7 is a new member of dendritic cell chemokine receptor that is up regulated upon maturation. AB - Dendritic cells (DC) that are stimulated with inflammatory mediators can maturate and migrate from nonlymphoid tissues to lymphoid organs to initiate T cell mediated immune responses. This migratory step is closely related to the maturation of the DC. In an attempt to identify chemokine receptors that might influence migration and are selectively expressed in mature DC, we have discovered that the chemokine receptor, EBI1/CCR7, is strikingly up-regulated upon maturation in three distinct culture systems: 1) mouse bone marrow-derived DC, 2) mouse epidermal Langerhans cells, and 3) human monocyte-derived DC. The EBI1/CCR7 expressed in mature DC is functional because ELC/MIP-3beta, recently identified as a ligand of EBI1/CCR7, induces a rise in intracellular free calcium concentrations and directional migration of human monocyte-derived mature DC (HLA DRhigh, CD1a(low), CD14-, CD25+, CD83+, and CD86high) in a dose-dependent manner, but not of immature DC (HLA-DRlow, CD1a(high), CD14-, CD25-, CD83-, and CD86-). In contrast, macrophage inflammatory protein-1alpha (MIP-1alpha), monocyte chemotactic protein-3 (MCP-3), and RANTES are active on immature DC but not on mature DC. Thus, it seems likely that MIP-1alpha, MCP-3, and RANTES can mediate the migration of immature DC located in peripheral sites, whereas ELC/MIP-3beta can direct the migration of Ag-carrying DC from peripheral inflammatory sites, where DC are stimulated to up-regulate the expression of EBI1/CCR7, to lymphoid organs. It is postulated that different chemokines and chemokine receptors are involved in DC migration in vivo, depending on the maturation state of DC. PMID- 9743377 TI - CXCR4 and CCR5 on human thymocytes: biological function and role in HIV-1 infection. AB - Thymocyte infection with HIV-1 is associated with thymic involution and impaired thymopoiesis, particularly in pediatric patients. To define mechanisms of thymocyte infection, we examined human thymocytes for expression and function of CXCR4 and CCR5, the major cell entry coreceptors for T cell line-tropic (T tropic) and macrophage-tropic (M-tropic) strains of HIV-1, respectively. CXCR4 was detected on the surface of all thymocytes. CXCR4 expression on mature, high level TCR thymocytes was similar to that on peripheral blood T cells, but was much lower than that on immature thymocytes, including CD34+ thymic progenitors. Consistent with this, stroma-derived factor-1 (SDF-1) induced calcium flux primarily in immature thymocytes, with CD34+ progenitors giving the strongest response. In addition, SDF-1 mRNA was detected in thymic-derived stromal cells, and SDF-1 induced chemotaxis of thymocytes, suggesting that CXCR4 may play a role in thymocyte migration. Infection of immature thymocytes by the T-tropic HIV-1 strain LAI was 10-fold more efficient than that in mature thymocytes, consistent with their relative CXCR4 surface expression. Anti-CXCR4 antiserum or SDF-1 blocked fusion of thymocytes with cells expressing the LAI envelope. In contrast to CXCR4, CCR5 was detected at low levels on thymocytes, and CCR5 agonists did not induce calcium flux or chemotaxis in thymocytes. However, CD4+ mature thymocytes were productively infected with the CCR5-tropic strain Ba-L, and this infection was specifically inhibited with the CCR5 agonist, macrophage inflammatory protein-1beta. Our data provide strong evidence that CXCR4 and CCR5 function as coreceptors for HIV-1 infection of human thymocytes. PMID- 9743378 TI - Tissue and T cell distribution of precursor and mature IL-16. AB - IL-16 is a novel cytokine, which is chemoattractant for CD4+ T cells, macrophages, and eosinophils. Recently, it was reported that IL-16 is synthesized as an approximately 80-kDa precursor molecule, pro-IL-16. Since little is known about the processing and tissue distribution of IL-16 and pro-IL-16, we investigated the distribution of IL-16 mRNA and protein in human lymphoid tissue. Northern blotting identified IL-16 mRNA predominantly in normal lymphoid organs, including PBMC, spleen, and thymus. Immunohistochemistry of human lymph node localized IL-16 protein to lymphocyte cytoplasm within T cell zones and occasionally in lymphocytes in B cell zones. Flow cytometric detection of intracellular IL-16 showed that >70% of CD4+ and CD8+ T cells constitutively expressed IL-16 protein. Western blot analysis of PBMC revealed nearly all of this protein to be approximately 80-kDa pro-IL-16 in unstimulated PBMC, and upon cell activation, the amino terminus of pro-IL-16 is processed into multiple fragments. These results show that pro-IL-16 is widely and constitutively expressed and suggest that the amino terminus of the protein can be processed upon cell activation. PMID- 9743379 TI - CD40-mediated signal transduction in human airway smooth muscle. AB - CD40 is a member of the TNF receptor family that was initially described on the surface of B cells. Recently, CD40 has also been described on mesenchymal cells, such as endothelial cells and fibroblasts, where engagement by its ligand CD40 ligand can lead to up-regulation of costimulatory and cell adhesion molecules, as well as secretion of proinflammatory cytokines. Since airway inflammation potentially involves cell-cell interactions of T cells and eosinophils (which express CD40 ligand) with airway smooth muscle (ASM) cells, we postulated that ASM may express CD40 and that engagement of ASM CD40 may modulate smooth muscle cell function. We demonstrate that CD40 is expressed on cultured human ASM and that expression can be increased by treatment with TNF-alpha or IFN-gamma. Cross linking CD40 on ASM resulted in enhanced IL-6 secretion and an increase in intracellular calcium concentrations, which were dependent on calcium influx. We show that CD40-mediated signaling events include protein tyrosine phosphorylation and activation of NF-kappaB. Pretreatment of ASM with the tyrosine kinase inhibitors genistein or herbimycin inhibited the rapid mobilization of calcium induced via CD40, suggesting that calcium mobilization was coupled to activation of protein tyrosine kinases. In addition, inhibition of calcium influx inhibited both CD40-mediated NF-kappaB activation and enhancement of IL-6 secretion. These results delineate a potentially important CD40-mediated signal-transduction pathway in ASM, involving protein tyrosine kinase-dependent calcium mobilization, NF-kappaB activation, and IL-6 production. Together, these results suggest a mechanism whereby T cell/smooth muscle cell interactions may potentiate airway inflammation. PMID- 9743380 TI - Th2-induced eotaxin expression and eosinophilia coexist with Th1 responses at the effector stage of lung inflammation. AB - The T cell-mediated lung inflammation that is associated with allergic asthma is characterized mainly by massive eosinophil infiltration, which induces airway injury and the subsequent late-phase reactivity. Because Th2 cells are often isolated from asthmatic subjects, these cells are postulated to play a role in asthma pathogenesis. We report that adoptively transferred, influenza hemagglutinin-specific Th1 and Th2 cells induced different patterns of chemokines leading to different types of cellular infiltration. Th2 cells were sufficient to induce dramatic Ag-dependent lung eosinophilia and eotaxin expression; by contrast, Th1 transfer primarily induced neutrophil recruitment with little eotaxin production. To determine whether Th1 cells show inhibitory effects on Th2 cell-mediated responses, Th1 and Th2 cells were cotransferred. Hemagglutinin specific Th1 cells did not inhibit Ag-induced lung eosinophilia, nor did they inhibit eotaxin expression. Furthermore, influenza virus infection of the lung in mice receiving hemagglutinin-specific Th2 cells also induced eotaxin expression and eosinophilia that could not be inhibited by the cotransfer of Th1 cells. Our results show that Th2-mediated allergic lung inflammation coexists with the Th1 mediated responses that are stimulated by diverse forms of Ags. PMID- 9743381 TI - TNFR80-dependent enhancement of TNFR60-induced cell death is mediated by TNFR associated factor 2 and is specific for TNFR60. AB - Costimulation of TNFR80 can strongly enhance TNFR60-induced cell death. In this study, we show that this enhancement is TNFR60 selective, as neither TNF-related apoptosis-inducing ligand/Apo2 ligand-, Apo1/Fas-, ceramide-, nor daunorubicin mediated cell death was affected by costimulation of TNFR80. We further demonstrate that TNFR-associated factor 2 (TRAF2) is critically involved in both negative and positive regulation of TNF-induced cell death. Overexpression of TRAF2 and of a TRAF2 mutant, deficient in nuclear factor-kappaB activation, selectively desensitized and enhanced, respectively, TNFR60-induced cell death in HeLa cells. However, upon costimulation of TNFR80, which mediates activation of nuclear factor-kappaB and the c-Jun amino-terminal kinase via TRAF2, TNF-induced cell death is drastically enhanced in parental and TRAF2-transfected, but not in TRAF2 (87-501)-transfected cells. These data point to a critical role of TRAF2 in the apoptotic TNFR cross talk, whereby the TNFR80-dependent enhancement of TNFR60 induced cell death is due to TNFR80-mediated negative regulation of TRAF2 function(s). An interference with TRAF2 function was confirmed independently by analysis of c-Jun amino-terminal kinase activation via TNFR60 upon prestimulation of TNFR80. We propose that the apoptotic TNFR cross talk is based on TNFR80 mediated abrogation of antiapoptotic TRAF2-dependent signaling pathways initiated by TNFR60, but not Apo1/Fas or the apoptotic TNF-related apoptosis-inducing ligand receptors. PMID- 9743382 TI - IL-12, but not IFN-gamma, plays a major role in sustaining the chronic phase of colitis in IL-10-deficient mice. AB - IL-10-deficient (IL-10(-/-)) mice develop chronic enterocolitis mediated by CD4+ Th1 cells producing IFN-gamma. Because IL-12 can promote Th1 development and IFN gamma production, the ability of neutralizing anti-IL-12 mAb to modulate colitis in IL-10(-/-) mice was investigated. Anti-IL-12 mAb treatment completely prevented disease development in young IL-10(-/-) mice. Treatment of adult mice resulted in significant amelioration of established disease accompanied by reduced numbers of mesenteric lymph node and colonic CD4+ T cells and of mesenteric lymph node T cells spontaneously producing IFN-gamma. In contrast, anti-IFN-gamma mAb had minimal effect on disease reversal, despite a significant preventative effect in young mice. These findings suggested that IL-12 sustains colitis by supporting the expansion of differentiated Th1 cells that mediate disease independently of their IFN-gamma production. This conclusion was supported by the finding that anti-IL-12 mAb greatly diminished the ability of a limited number of CD4+ T cells expressing high levels of CD45RB from diseased IL 10(-/-) mice to expand and cause colitis in recombination-activating gene-2(-/-) recipients, while anti-IFN-gamma mAb had no effect. Furthermore, IL-12 could support pathogenic IL-10(-/-) T cells stimulated in vitro in the absence of IL-2. While these studies show that IL-12 plays an important role in sustaining activated Th1 cells during the chronic phase of disease, the inability of anti-IL 12 mAb to abolish established colitis or completely prevent disease transfer by Thl cells suggests that additional factors contribute to disease maintenance. PMID- 9743383 TI - Signaling through a CD3 gamma-deficient TCR/CD3 complex in immortalized mature CD4+ and CD8+ T lymphocytes. AB - The biologic role of each CD3 chain and their relative contribution to the signals transduced through the TCR/CD3 complex and to downstream activation events are still controversial: they may be specialized or redundant. We have immortalized peripheral blood CD4+ and CD8+ T lymphocytes from a human selective CD3 gamma deficiency using Herpesvirus saimiri. The accessibility of the mutant TCR/CD3 complex to different Abs was consistently lower in immortalized CD8+ cells when compared with CD4+ cells, relative to their corresponding CD3 gamma sufficient controls. Several TCR/CD3-induced downstream activation events, immediate (calcium flux), early (cytotoxicity and induction of surface CD69 or CD40L activation markers or intracellular TNF-alpha) and late (proliferation and secretion of TNF-alpha), were normal in gamma-deficient cells, despite the fact that their TCR/CD3 complexes were significantly less accessible than those of controls. In contrast, the accumulation of intracellular IL-2 or its secretion after CD3 triggering was severely impaired in gamma-deficient cells. The defect was upstream of protein kinase C activation because addition of transmembrane stimuli (PMA plus calcium ionophore) completely restored IL-2 secretion in gamma deficient cells. These results suggest that the propagation of signals initiated at the TCR itself can result in a modified downstream signaling cascade with distinct functional consequences when gamma is absent. They also provide evidence for the specific participation of the CD3 gamma chain in the induction of certain cytokine genes in both CD4+ and CD8+ human mature T cells. These immortalized mutant cells may prove to be useful in isolating cytosolic signaling pathways emanating from the TCR/CD3 complex. PMID- 9743384 TI - High resolution mapping of the B cell epitopes of staphylokinase in humans using negative selection of a phage-displayed antigen library. AB - Staphylokinase (Sak), a 16-kDa protein secreted by Staphylococcus aureus, induces fibrin-specific thrombolysis in patients with thrombotic disorders. However, Sak also elicits high titers of neutralizing Abs that persist for several months and preclude its repeated use in humans. To identify the antigenic determinants of Sak recognized by humans, a phage-displayed library of Sak variants was selected for mutants that escape binding to an affinity matrix derivatized with patient specific polyclonal anti-Sak Abs. Fifty-six escape Sak variants were identified after three selection cycles using human polyclonal anti-Sak IgGs obtained from four different patients. DNA sequencing revealed 213 amino acid substitutions, of which 73% were found at 25 positions clustered in eight discontinuous Sak antigenic segments. Although each antigenic segment was recognized to a variable extent by each patient antiserum, the main epitopes of Sak in all patients were roughly targeted to two large discontinuous areas covering 35% of the solvent accessible surface of Sak. The antigenic area I comprises three segments centered on residues 66, 73, and 135, while the antigenic area II consists of four segments centered on positions 20, 95, 102, and 121. These results suggest that a secondary immune response against Sak can occur in patients, and confirm an initial site-directed mutagenesis study wherein amino acid Lys74 was shown to play a prominent antigenic role. Comprehensive mapping of the most relevant sites of Sak that are antigenic for humans will guide efforts to modulate the immunogenicity of this therapeutically important molecule. PMID- 9743385 TI - Different molecular events result in low protein levels of mannan-binding lectin in populations from southeast Africa and South America. AB - Previous studies have shown that three point mutations in exon 1 and a particular promoter haplotype of the mannan-binding lectin (MBL) gene lead to a dramatic decrease in the serum concentration of MBL. In this study, MBL genotypes and serum concentrations were determined in unrelated individuals in a population from Mozambique (n = 154) and in two native Indian tribes from Argentina (i.e., the Chiriguanos (n = 43) and the Mapuches (n = 25)). In both populations, the MBL concentrations were low compared with those found in Eskimo, Asian, and European populations. In Africans, the low serum concentrations were due to a high allele frequency (0.24) of the codon 57 (C) variant, which resulted in a high frequency of individuals with MBL deficiency (0.06), and were also due to the effect of a relatively high frequency (0.13) of low-producing promoter haplotypes. The low concentrations in the South American populations were primarily due to an extremely high allele frequency of the codon 54 (B) variant in both the Chiriguanos (0.42) and the Mapuches (0.46), resulting in high frequencies of individuals with MBL deficiency (0.14 and 0.16, respectively). In the search for additional genetic variants, we found five new promoter mutations that might help to elucidate the evolution of the MBL gene. Taken together, the results of this study show that different molecular mechanisms are the basis for low MBL levels on the two continents. PMID- 9743386 TI - Monoclonal antibody therapy of B cell lymphoma: signaling activity on tumor cells appears more important than recruitment of effectors. AB - Despite the recent success of mAb in the treatment of certain malignancies, there is still considerable uncertainty about the mechanism of action of anti-cancer Abs. Here, a panel of rat anti-mouse B cell mAb, including Ab directed at surface IgM Id, CD19, CD22, CD40, CD74, and MHC class II, has been investigated in the treatment of two syngeneic mouse B cell lymphomas, BCL1 and A31. Only three mAb were therapeutically active in vivo, anti-Id, anti-CD19, and anti-CD40. mAb to the other Ags showed little or no therapeutic activity in either model despite giving good levels of surface binding and activity in Ag-dependent cellular cytotoxicity and complement assays, and in some cases inhibiting cell growth in vitro. We conclude that the activity of mAb in vitro does not predict therapeutic performance in vivo. Furthermore, in vivo tracking experiments using fluorescently tagged cells showed that anti-Id and anti-CD40 mAb probably operate via different mechanisms: the anti-Id mAb cause growth arrest that is almost immediate and does not eliminate cells over a period of 5 or 6 days, and the anti CD40 mAb have a delayed effect that allows tumor to grow normally for 3 days, but then abruptly eradicates lymphoma cells. This work supports the belief that mAb specificity is critical to therapeutic success in lymphoma and that, in addition to any effector-recruiting activity they may possess, in vivo mAb operate via mechanisms that involve cross-linking and signaling of key cellular receptors. PMID- 9743387 TI - Generation of human cytolytic T lymphocyte lines directed against prostate specific antigen (PSA) employing a PSA oligoepitope peptide. AB - Prostate-specific Ag (PSA), which is expressed in a majority of prostate cancers, is a potential target for specific immunotherapy. Previous studies have shown that two 10-mer PSA peptides (designated PSA-1 and PSA-3) selected to conform to human HLA class I-A2 motifs can elicit CTL responses in vitro. A longer PSA peptide (30-mer) designated PSA-OP (oligoepitope peptide), which contains both the PSA-1 and PSA-3 HLA-A2 epitopes and an additional potential CTL epitope (designated PSA-9) for the HLA-class I-A3 allele, was investigated for the ability to induce cytotoxic T cell activity. T cell lines from different HLA-A2 and HLA-A3 donors were established by in vitro stimulation with PSA-OP; the CTL lines lysed PSA-OP as well as PSA-1- or PSA-3-pulsed C1R-A2 cells, and PSA-OP and PSA-9-pulsed C1R-A3 cells, respectively. The CTL lines derived from the PSA-OP peptide also lysed PSA-positive prostate cancer cells. PSA-OP-derived T cell lines also lysed recombinant vaccinia-PSA-infected targets but not targets infected with wild-type vaccinia. PSA-OP did not bind HLA-A2 and HLA-A3 molecules. The decrease in cytotoxicity in the presence of protease inhibitors suggests that the PSA-OP is cleaved into shorter peptides, which in turn can interact with HLA-class I molecules and, as a consequence, induce CTL-mediated lysis. We have also demonstrated that it is possible to induce CTL responses in HLA-A2.1/Kb transgenic mice by immunization with PSA-OP with adjuvant. These studies thus provide evidence that oligopeptides such as PSA-OP may be useful candidates for peptide-based cancer vaccines. PMID- 9743388 TI - Expression of chemokine receptors CXCR4 and CCR5 in HIV-1-infected and uninfected individuals. AB - The chemokine receptors CXCR4 and CCR5 have been identified as major coreceptors for HIV-1 entry into CD4+ T cells. The majority of primary HIV-1 isolates in early disease use CCR5 as a coreceptor, whereas during disease progression with the emergence of syncytium-inducing viruses, CXCR4 is also used. We performed a cross-sectional study in which we evaluated the expression of two HIV-1 coreceptors, CCR5 and CXCR4, in whole blood samples taken from HIV-1-infected and uninfected individuals. We demonstrate that CXCR4 on CD4+ and CD8+ T cells, and CD14+ monocytes is significantly down-regulated, and CCR5 expression on CD4+ T cells is up-regulated in HIV-infected individuals compared with uninfected controls. Coreceptor expression correlated with the level of cellular activation in vivo in both HIV-infected and uninfected individuals, with CXCR4 being expressed predominantly on quiescent (HLA-DR-) T cells and CCR5 being expressed predominantly on activated (HLA-DR+) T cells. Lower expression of CXCR4 and higher expression of CCR5 on CD4+ T cells correlated with advancing disease. In addition, a tendency for greater activation of CXCR4+CD4+ T cells in patients with advanced disease was observed. Patients who harbored syncytium-inducing viruses, however, could not be distinguished from those who harbored nonsyncytium inducing viruses based on the level of CD4+ T cell activation or chemokine receptor expression. PMID- 9743389 TI - Does size matter? PMID- 9743390 TI - Receptor-specific reversible sedation: beginning of new era of anesthesia? PMID- 9743391 TI - Remifentanil pharmacokinetics in obese versus lean patients. AB - BACKGROUND: Remifentanil is a short-acting opioid whose pharmacokinetics have been characterized in detail. However, the impact of obesity on remifentanil pharmacokinetics has not been specifically examined. The goal of this study was to investigate the influence of body weight on remifentanil pharmacokinetics. METHODS: Twelve obese and 12 matched lean subjects undergoing elective surgery received a 1-min remifentanil infusion after induction of anesthesia. Arterial blood samples were collected for determination of remifentanil blood concentrations. Each subject's pharmacokinetic parameters were estimated by fitting a two-compartment model to the concentration versus time curves. Nonlinear mixed-effects population models examining the influence of lean body mass (LBM) and total body weight (TBW) were also constructed. Clinical simulations using the final population model were performed. RESULTS: The obese patient cohort reached substantially higher remifentanil concentrations. The individual pharmacokinetic parameters of a two-compartment model were not significantly different between the obese versus lean cohorts (unless normalized to TBW). The final population model scaled central clearance and the central and peripheral distribution volumes to LBM. The simulations illustrated that remifentanil pharmacokinetics are not grossly different in obese versus lean subjects and that TBW based dosing in obese patients can result in excessively high remifentanil concentrations. CONCLUSIONS: The essential findings of the study are that remifentanil's pharmacokinetics are not appreciably different in obese versus lean subjects and that remifentanil pharmacokinetic parameters are therefore more closely related to LBM than to TBW. Clinically this means that remifentanil dosing regimens should be based on ideal body weight (or LBM) and not TBW. PMID- 9743392 TI - Reversal of the sedative and sympatholytic effects of dexmedetomidine with a specific alpha2-adrenoceptor antagonist atipamezole: a pharmacodynamic and kinetic study in healthy volunteers. AB - BACKGROUND: Specific and selective alpha2-adrenergic drugs are widely exploited in veterinary anesthesiology. Because alpha2-agonists are also being introduced to human practice, the authors studied reversal of a clinically relevant dexmedetomidine dose with atipamezole, an alpha2-antagonist, in healthy persons. METHODS: The study consisted of two parts. In an open dose-finding study (part 1), the intravenous dose of atipamezole to reverse the sedative effects of 2.5 microg/kg of dexmedetomidine given intramuscularly was determined (n = 6). Part 2 was a placebo-controlled, double-blinded, randomized cross-over study in which three doses of atipamezole (15, 50, and 150 microg/kg given intravenously in 2 min) or saline were administered 1 h after dexmedetomidine at 1-week intervals (n = 8). Subjective vigilance and anxiety, psychomotor performance, hemodynamics, and saliva secretion were determined, and plasma catecholamines and serum drug concentrations were measured for 7 h. RESULTS: The mean +/- SD atipamezole dose needed in part 1 was 104+/-44 microg/kg. In part 2, dexmedetomidine induced clear impairments of vigilance and psychomotor performance that were dose dependently reversed by atipamezole (P < 0.001). Complete resolution of sedation was evident after the highest (150 microg/kg) dose, and the degree of vigilance remained high for 7 h. Atipamezole dose dependently reversed the reductions in blood pressure (P < 0.001) and heart rate (P = 0.009). Changes in saliva secretion and plasma catecholamines were similarly biphasic (i.e., they decreased after dexmedetomidine followed by dose-dependent restoration after atipamezole). Plasma norepinephrine levels were, however, increased considerably after the 150 microg/kg dose of atipamezole. The pharmacokinetics of atipamezole were linear, and elimination half-lives for both drugs were approximately 2 h. Atipamezole did not affect the disposition of dexmedetomidine. One person had symptomatic sinus arrest, and another had transient bradycardia approximately 3 h after receiving dexmedetomidine. CONCLUSIONS: The sedative and sympatholytic effects of intramuscular dexmedetomidine were dose dependently antagonized by intravenous atipamezole. The applied infusion rate (75 microg x kg(-1) x min(-1)) for the highest atipamezole dose was, however, too fast, as evident by transient sympathoactivation. Similar elimination half-lives of these two drugs are a clear advantage considering the possible clinical applications. PMID- 9743393 TI - Epidural bupivacaine-morphine analgesia versus patient-controlled analgesia following abdominal aortic surgery: analgesic, respiratory, and myocardial effects. AB - BACKGROUND: The efficacy and effects of epidural analgesia compared with patient controlled analgesia (PCA) have not been reported in patients undergoing major vascular surgery. We compared the effects of epidural bupivacaine-morphine with those of intravenous PCA morphine after elective infrarenal aortic surgery. METHODS: Forty patients classified as American Society of Anesthesiologists physical status 2 or 3 received general anesthesia plus postoperative PCA using morphine sulfate (group PCA; n = 21) or general anesthesia plus perioperative epidural morphine-bupivacaine (group EPI; n = 19) during a period of 48 h. During operation, EPI patients received 0.05 mg/kg epidural morphine and 5 ml 0.25% bupivacaine followed by an infusion of 0.125% bupivacaine with 0.1% morphine (0.1 mg/ ml); group PCA received 0.1 mg/kg intravenous morphine sulfate. Continuous electrocardiographic monitoring (V4 and V5 leads) was performed from the night before surgery until 48 h afterward. Respiratory inductive plethysmographic data were recorded after tracheal extubation. Visual analog pain scores at rest and after movement were performed every 4 h after extubation. RESULTS: Nurse administered intravenous morphine and time to tracheal extubation were less in group EPI, as were visual analog pain scores at rest and after movement from 20 to 48 h. Complications and the duration of intensive care unit and hospital stay were comparable. There was a similar, low incidence of postoperative apneas, slow respiratory rates, desaturation, and S-T segment depression. CONCLUSIONS: Epidural morphine-bupivacaine is associated with reduced early postoperative intravenous opioid requirements, more rapid tracheal extubation, and superior analgesia after abdominal aortic surgery, with comparable respiratory effects. PMID- 9743394 TI - The effect of adding a minidose of clonidine to intrathecal sufentanil for labor analgesia. AB - BACKGROUND: Preliminary studies have suggested that the addition of clonidine to intrathecal sufentanil prolongs analgesia without producing motor blockade. METHODS: Fifty-three nulliparous women in painful labor were included in this prospective, randomized, double-blinded study. Parturients at 2- to 5-cm cervical dilation received either 5 microg sufentanil plus 30 microg clonidine or 5 microg sufentanil intrathecally, followed by 5 mg bupivacaine epidurally. The primary outcome was time until first request for additional analgesia. Visual analog pain scores, sensory changes, blood pressure, heart rate, ephedrine requirements, motor blockade, sedation, pruritus, and nausea were also recorded. RESULTS: All parturients but one had effective analgesia in both groups, with similar sensory levels never exceeding T2. The duration (mean +/- SD) of analgesia was longer in the sufentanil-clonidine group: 125+/-46 versus 97+/-30 min (P = 0.007). The incidence of hypotension and the ephedrine requirements (median with range) were higher in the sufentanil-clonidine group: 63% versus 12% (P < 0.001) and 7.5 mg [range, 0-25.5 mg] versus 0 mg [range, 0-6 mg] (P < 0.0001). The incidence of fetal heart rate abnormalities during the first 30 min after intrathecal injection was similar in both groups (17% vs. 19%). No parturient had motor blockade. CONCLUSIONS: The addition of 30 microg clonidine to 5 microg intrathecal sufentanil extended the duration of labor analgesia without producing motor blockade. However, as previously reported with 100-200 microg clonidine, the incidence of hypotension and the ephedrine requirements were also increased, even when 30 microg clonidine only was added. PMID- 9743395 TI - Acute depression of myocardial beta-adrenergic receptor signaling during cardiopulmonary bypass: impairment of the adenylyl cyclase moiety. Duke Heart Center Perioperative Desensitization Group. AB - BACKGROUND: Previously the authors showed that myocardial beta-adrenergic (betaAR) function is reduced after cardiopulmonary bypass (CPB) in a canine model Whether CPB results in similar effects on betaAR function in adult humans is not known. Therefore the current study tested two hypotheses: (1) That myocardial betaAR signaling is reduced in adult humans after CPB, and (2) that administration of long-term preoperative betaAR antagonists prevents this process. METHODS: After they gave informed consent, 52 patients undergoing aortocoronary surgery were enrolled. Atrial biopsies were obtained before CPB and immediately before discontinuation of CPB. Plasma catecholamine concentrations, myocardial betaAR density, and functional responsiveness (basal, isoproterenol, zinterol, sodium fluoride, and manganese-stimulated adenylyl cyclase activity) were assessed. RESULTS: Catecholamine levels increased significantly during CPB (P < 0.005). Myocardial betaAR adenylyl cyclase coupling decreased during CPB, as evidenced by a 21% decrease in isoproterenol-stimulated adenylyl cyclase activity (750 [430] pmol cyclic adenosine monophosphate per milligram total protein 15 min before CPB compared with 540 [390] at the end of CPB, P = 0.0062, medians [interquartile range]) despite constant betaAR density. Differential activation along the betaAR signal transduction cascade localized the defect to the adenylyl cyclase moiety. Administration of long-term preoperative betaAR antagonists did not prevent acute CPB-induced myocardial betaAR dysfunction. CONCLUSIONS: These data indicate that the myocardial adenylyl cyclase response to betaAR agonists decreases acutely in adults during aortocoronary surgery requiring CPB, regardless of whether long-term preoperative betaAR antagonists are administered. The mechanism underlying acute betaAR dysfunction appears to be direct impairment of the adenylyl cyclase moiety. Similar increases in manganese-stimulated activity before and at the end of CPB show preserved adenylyl cyclase catalytic activity, suggesting that other mechanisms (such as decreased protein levels or altered isoform expression or function) may be responsible for decreased adenylyl cyclase function. PMID- 9743396 TI - Effects of alfentanil on the ventilatory response to sustained hypoxia. AB - BACKGROUND: The ventilatory response to acute hypoxia is biphasic, with an initial rapid increase followed by a slower decline. In humans, there is evidence that the magnitude of the decline in ventilation is proportional to the size of the initial increase. This study was done to define the role of exogenous opioids in the ventilatory decline seen with prolonged hypoxia. METHODS: Ten healthy persons were exposed to isocapnic hypoxia for 25 min, followed by 5 min of isocapnic normoxia and 5 min of isocapnic hypoxia. These conditions were repeated during a computer-controlled alfentanil infusion. RESULTS: Serum alfentanil levels were constant among the volunteers (38+/-12 ng/ml). Alfentanil decreased both the initial and second acute hypoxic responses (from 1.27+/-0.73 to 0.99+/ 0.39 l x min(-1) x %(-1), P < 0.05; and from 0.99+/-0.70 to 0.41+/-0.29 l x min( 1) x %(-1), P < 0.05, respectively). The magnitude of the decrease in ventilation during the 25 min of hypoxia was not changed (10+/-3.3 l/min for control; 12.3+/ 7.5 l/min for alfentanil). CONCLUSIONS: Alfentanil reduced the acute ventilatory response to hypoxia. The absolute value of hypoxic ventilatory decline was not increased, but a measure of residual hypoxic ventilatory decline (the ratio of ventilation between the second and first steps into hypoxia) was decreased, which supports the hypothesis that opioids potentiate centrally mediated ventilatory decline. PMID- 9743397 TI - Tryptase levels are not increased during vancomycin-induced anaphylactoid reactions. AB - BACKGROUND: Anaphylaxis, mediated by immunoglobulin E, may be clinically indistinguishable but is mechanistically different than chemically mediated anaphylactoid reactions induced by drugs such as morphine, curare, and vancomycin. A test to distinguish anaphylactic from anaphylactoid reactions would clarify therapeutic and medicolegal issues. Tryptase levels identify anaphylactic reactions but have not been evaluated in vivo during anaphylactoid reactions. A prospective, randomized, double-blinded, placebo-controlled trial of antihistamine chemoprophylaxis for rapid vancomycin infusion was performed, and plasma tryptase was measured using a new immunoassay. Histamine release was established by measurement of plasma histamine and the ability of prophylactic H1 and H2 antagonists to prevent common histamine-associated side effects. Tryptase levels were compared with histamine levels and clinical symptoms. METHODS: Before elective arthroplasty, 40 patients received vancomycin infusion (1 g over 10 min) and pretreatment with either antihistamines (1 mg/kg diphenhydramine and 4 mg/kg cimetidine) or placebo. Changes in tryptase (at peak histamine and 10 min after vancomycin infusion), histamine levels, and histamine-mediated symptoms were assessed using Fisher's exact test, the Student's t test, or the paired t test, as appropriate. Logistic regression models were used to quantify the association of clinical symptoms with antihistamine treatment and serum levels. RESULTS: Plasma tryptase levels were unchanged (99% CI, -0.5 to 1.6) independent of increased histamine levels, antihistamine pretreatment, clinical symptoms, or all of these. Histamine levels >1 ng/ml were significantly associated with hypotension, moderate-to-severe rash, and stopped infusion. Antihistamine pretreatment significantly decreased the incidence and severity of the reactions. CONCLUSION: Plasma tryptase levels were not significantly elevated in confirmed anaphylactoid reactions, so they can be used to distinguish chemical from immunologic reactions. PMID- 9743398 TI - Dose-dependent reduction of the minimum local analgesic concentration of bupivacaine by sufentanil for epidural analgesia in labor. AB - BACKGROUND: The minimum local analgesic concentration (MLAC) has been defined as the median effective local analgesic concentration in a 20-ml volume for epidural analgesia in the first stage of labor. The aim of this study was to determine the local anesthetic-sparing efficacy of epidural sufentanil by its effect on the MLAC of bupivacaine. METHODS: In this double-blind, randomized, prospective study, 147 parturients at < or = 7 cm cervical dilation who requested epidural analgesia were allocated to one of four study groups. After a lumbar epidural catheter was placed, study participants received 20 ml bupivacaine (n = 38), bupivacaine with sufentanil 0.5 microg/ml (n = 38), bupivacaine with sufentanil 1 microg/ml (n = 33), or bupivacaine with sufentanil 1.5 microg/ml (n = 38). The concentration of bupivacaine was determined by the response of the previous patient using up-down sequential allocation. The analgesic efficacy was assessed using 100-mm visual analog pain scores, with < or = 10 mm within 30 min defined as effective. RESULTS: The MLAC of bupivacaine alone was 0.104% wt/vol (95% CI, 0.090-0.117). The addition of sufentanil at doses of 0.5 microg/ml, 1 microg/ml, and 1.5 microg/ml resulted in significant reductions (P < 0.0001) in the MLAC of bupivacaine to 0.048% wt/vol (95% CI, 0.030- 0.065), 0.021% wt/vol (95% CI, 0 0.055), and 0.009% wt/vol (95% CI, 0-0.023), respectively. CONCLUSIONS: This study showed a significant (P < 0.0001) dose-dependent reduction in the MLAC ofbupivacaine by sufentanil. PMID- 9743399 TI - Transient neurologic symptoms after spinal anesthesia: an epidemiologic study of 1,863 patients. AB - BACKGROUND: Recent evidence suggests that transient neurologic symptoms commonly follow lidocaine spinal anesthesia. However, information concerning factors that affect their occurrence is limited. Accordingly, to evaluate many potential risk factors, the authors undertook a prospective, multicenter, epidemiologic study. METHODS: On a voluntary basis, anesthetists at 15 participating centers forwarded a data sheet on patients who had spinal anesthesia to a research nurse blinded to the details of anesthesia and surgery. A subset was randomly selected for follow up. The pressure [corrected] of transient neurologic symptoms, defined as leg or buttock pain, was the principal outcome variable. Logistic regression was used to control for potential confounders, and adjusted odds ratios and confidence intervals were used to estimate relative risk. RESULTS: During a 14-month period, 1,863 patients were studied, of whom 47% received lidocaine, 40% bupivacaine, and 13% tetracaine. Patients given lidocaine were at higher risk for symptoms compared with those receiving bupivacaine (relative risk, 5.1; 95% CI, 2.5 to 10.2) or tetracaine (relative risk, 3.2; 95% CI, 1.04 to 9.84). For patients who received lidocaine, the relative risk of transient neurologic symptoms was 2.6 (95% CI, 1.5 to 4.5) with the lithotomy position compared with other positions, 3.6 (95% CI, 1.9 to 6.8), for outpatients compared with inpatients, and 1.6 (95% CI, 1 to 2.5) for obese (body mass index >30) compared with nonobese patients. CONCLUSIONS: These results indicate that transient neurologic symptoms commonly follow lidocaine spinal anesthesia but are relatively uncommon with bupivacaine or tetracaine. The data identify lithotomy position and outpatient status as important risk factors in patients who receive lidocaine. Among other factors postulated to increase risk, obesity had an effect of borderline statistical significance, whereas age, sex, history of back pain, needle type, and lidocaine dose and concentration failed to affect risk. PMID- 9743400 TI - Diphenylhydramine increases ventilatory drive during alfentanil infusion. AB - BACKGROUND: Diphenhydramine is used as an antipruritic and antiemetic in patients receiving opioids. Whether it might exacerbate opioid-induced ventilatory depression has not been determined. METHODS: The ventilatory response to carbon dioxide during hyperoxia and the ventilatory response to hypoxia during hypercapnia (end-tidal pressure of carbon dioxide [PETCO2] is approximately equal to 54 mmHg) were determined in eight healthy volunteers. Ventilatory responses to carbon dioxide and hypoxia were calculated at baseline and during an alfentanil infusion (estimated blood levels approximately equal to 10 ng/ml) before and after diphenhydramine 0.7 mg/kg. RESULTS: The slope of the ventilatory response to carbon dioxide decreased from 1.08+/-0.38 to 0.79+/-0.36 l x min(-1) x mmHg( 1) (x +/- SD, P < 0.05) during alfentanil infusion; after diphenhydramine, the slope increased to 1.17+/-0.28 l x min(-1) x mmHg(-1) (P < 0.05). The minute ventilation (VE) at PETCO2 approximately equal to 46 mmHg (VE46) decreased from 12.1+/-3.7 to 9.7+/-3.6 l/min (P < 0.05) and the VE at 54 mmHg (VE54) decreased from 21.3+/-4.8 to 16.6+/-4.7 l/min during alfentanil (P < 0.05). After diphenhydramine, (VE46 did not change significantly, remaining lower than baseline at 9.9+/-2.9 l/min (P < 0.05), whereas VE54 increased significantly to 20.5+/-3.0 l/min. During hypoxia, VE at SpO2 = 90% (VE90) decreased from 30.5+/ 9.7 to 23.1+/-6.9 l/min during alfentanil (P < 0.05). After diphenhydramine, the increase in VE90 to 27.2+/-9.2 l/min was not significant (P = 0.06). CONCLUSIONS: Diphenhydramine counteracts the alfentanil-induced decrease in the slope of the ventilatory response to carbon dioxide. However, at PETCO2 = 46 mmHg, it does not significantly alter the alfentanil-induced shift in the carbon dioxide response curve. In addition, diphenhydramine does not exacerbate the opioid-induced depression of the hypoxic ventilatory response during moderate hypercarbia. PMID- 9743401 TI - Paralysis only slightly reduces the febrile response to interleukin-2 during isoflurane anesthesia. AB - BACKGROUND: Fever sometimes occurs during anesthesia. However, it is rare considering how often pyrogenic causes are likely to be present and how common fever is after surgery. This low incidence results in part from dose-dependent inhibition of fever by volatile anesthetics. Paralysis, however, may contribute by preventing shivering and the associated increase in metabolic heat production. Therefore the authors tested the hypothesis that paralysis during anesthesia decreases the febrile response to pyrogen administration. METHODS: Seven volunteers each participated on two study days. They were given 30 IU/g intravenous interleukin-2, followed 90 min later by an additional 70 IU/g dose. Anesthesia was induced 30 min after the second dose and maintained for 6 h with 0.6 minimum alveolar concentration isoflurane. The volunteers were randomly assigned to (1) paralysis with vecuronium or (2) no muscle relaxants. Body heat content and distribution were determined from measured tissue and skin temperatures. Data are presented as mean +/- SD; P < 0.05 was considered significant. RESULTS: There was no clinically important difference in peak core (tympanic membrane) temperatures on the unparalyzed (37.6+/-0.9 degrees C) and paralyzed (37.2+/-0.6 degrees C) days. Core heat content increased 1.2+/-0.7 kcal/kg over the last 5 h of anesthesia on the unparalyzed day, but only by 0.9+/ 0.4 kcal/kg when the volunteers were paralyzed. Peripheral tissue heat content increased 0.1+/-1.1 kcal/kg on the unparalyzed day but decreased 1.1+/-0.7 kcal/kg when the volunteers were paralyzed. Consequently, body heat content increased 1.3+/-1.3 kcal/kg on the unparalyzed day but decreased significantly by 0.2+/-0.8 kcal/kg when the volunteers were paralyzed. CONCLUSIONS: Paralysis prevented shivering from increasing the metabolic rate. Consequently, body heat content decreased during paralysis, whereas otherwise it increased. Thermoregulatory vasoconstriction was nonetheless able to maintain similar peak and integrated core temperatures on each study day. Administration of muscle relaxants thus is not the primary explanation for the relative paucity of intraoperative fever. PMID- 9743402 TI - Experimental determination of heat flow parameters during induction of general anesthesia. AB - BACKGROUND: Alterations in body temperature result from changes in tissue heat content. Heat flow is a complex function of vasomotor status and core, peripheral, and ambient temperatures. Consequently it is difficult to quantify specific mechanisms responsible for observed changes in body heat distribution. Therefore the authors developed two mathematical models that independently express regional tissue heat production and the motion of heat through tissues in terms of measurable quantities. METHODS: The equilibrium model expresses the effective regional heat transfer coefficient in terms of cutaneous heat flux, skin temperature, and temperature at the center of the extremity. It applies at steady states and provides a ratio of the heat transfer coefficients before and after an intervention. In contrast, the heat flow model provides a time-dependent estimate of the heat transfer coefficient in terms of ambient temperature, skin temperature, and temperature at the center of the extremity. RESULTS: Each model was applied to data acquired in a previous evaluation of heat balance during anesthesia induction. The relation between the ratio of steady state regional heat transfer coefficients calculated using each model was linear. The effective heat transfer coefficient for the forehead (a core site) decreased approximately 20% after induction of anesthesia. In contrast, heat transfer coefficients in the six tested extremity sites more than doubled. CONCLUSIONS: Effective heat transfer coefficients can be used to evaluate the thermal effects of various clinical interventions, such as induction of regional anesthesia or administration of vasodilating drugs. The heat transfer coefficient for the forehead presumably decreased because general anesthesia reduces brain perfusion. In contrast, increased heat transfer coefficients in the extremity sites indicate that thermoregulatory and anesthetic-induced vasodilation more than doubles the core-to-peripheral flow of heat. This flow of heat causes redistribution hypothermia, which is usually the major cause of core hypothermia during anesthesia. PMID- 9743403 TI - Heart rate variability during chemical thoracic sympathectomy. AB - BACKGROUND: Chemical thoracic sympathectomy (CTS) resulted in profound bradycardia in a patient with severe post-therapeutic neuralgia. To clarify the cause of this bradycardia, the authors evaluated heart rate variability using a Poincare plot, which is a scatter diagram of the current R-R interval plotted against the R-R interval immediately preceding it, in this patient and in others scheduled for CTS or mandibular block (MB). METHODS: Twenty-three patients were scheduled for CTS (n = 13, CTS group) and for MB (n = 10, MB group). Heart rate and the SD of the R-R interval variabilities spreading along the x axis (SDRR) and perpendicularly along the diagonal line of the Poincare plot (SDdeltaRR) were evaluated before, just after, and 1 h after the block. RESULTS: Neither group had significant changes in heart rate. The MB group showed no significant change in the SD(RR):SDdeltaRR ratio. In the CTS group, however, the SD(RR):SDdeltaRR ratio decreased significantly from 1.72+/-0.20 to 1.23+/-0.11 just after CTS. The previous patient, who had a high SD(RR):SDdeltaRR ratio of 3.45 before CTS, exhibited severe bradycardia (22 beats/min). CONCLUSIONS: The SD(RR):SDdeltaRR ratio decreased after CTS without any significant concomitant change in heart rate. The decrease in the SD(RR):SDdeltaRR. ratio indicates a reduction of cardiac sympathetic activity. However, CTS in patients having high SD(RR):SDdeltaRR ratios can result in profound bradycardia. PMID- 9743404 TI - Bispectral EEG index during nitrous oxide administration. AB - BACKGROUND: Nitrous oxide (N2O) is a commonly used sedative for painful diagnostic procedures and dental work. The authors sought to characterize the effects of N2O on quantitative electroencephalographic (EEG) variables including the bispectral index (BIS), a quantitative parameter developed to correlate with the level of sedation induced by a variety of agents. METHODS: Healthy young adult volunteers (n = 13) were given a randomized sequence of N2O/O2 combinations via face mask. Five concentrations of N2O (10, 20, 30, 40, and 50% atm) were administered for 15 min (20 min for the first step). EEG was recorded from bilateral frontal poles continuously. At the end of each exposure, level of sedation was assessed using primarily the Observer Assessment of Alertness/Sedation (OAA/S) scale. RESULTS: One subject withdrew from the study because of emesis at 50% N2O. N2O (50%) increased theta, beta, 40-50 Hz, and 70 110 Hz band powers. BIS and spectral edge frequency during 50% N2O/O2 did not differ significantly from baseline values. Abrupt decreases from higher to lower concentrations frequently evoked a profound, transient slowing of activity. No significant change in OAA/S was detected during the study. CONCLUSIONS: Although the spectral content of the EEG changed during N2O administration, reflecting some pharmacologic effect, the subjects remained cooperative and responsive throughout, and therefore N2O can only be considered a weak sedative at the tested concentrations. Despite changes in the lower and higher frequency ranges of EEG activity, the BIS did not change, which is consistent with its design objective as a specific measure of hypnosis. PMID- 9743405 TI - Formulation-dependent brain and lung distribution kinetics of propofol in rats. AB - BACKGROUND: Propofol when administered by brief infusion in a lipid-free formulation has a slower onset, prolonged offset and greater potency compared with an emulsion formulation. To understand these findings the authors examined propofol brain and lung distribution kinetics in rats. METHODS: Rats were infused with equieffective doses of propofol in emulsion (n = 21) or lipid-free formulation (n = 21). Animals were sacrificed at various times to harvest brain and lung. Arterial blood was sampled repeatedly from each animal until sacrifice. Deconvolution and moment analysis were used to calculate the half-life for propofol brain turnover (BT) and brain:plasma partition coefficient (Kp). Lung concentration-time profiles were compared for the two formulations. RESULTS: Peak propofol plasma concentrations for the lipid-free formulation were 50% of that observed for emulsion formulation, whereas peak lung concentrations for lipid free formulation were 300-fold higher than emulsion formulation. Brain Kp calculated from tissue disposition curve and ratio of brain:plasma area under the curves were 8.8 and 13, and 7.2 and 9.1 for emulsion and lipid-free formulations, respectively. BT were 2.4 and 2.5 min for emulsion and lipid-free formulations, respectively. CONCLUSIONS: Significant pulmonary sequestration and slow release of propofol into arterial circulation when administered in lipid-free vehicle accounts for the lower peak arterial concentration and sluggish arterial kinetics relative to that observed with the emulsion formulation. Higher Kp for the lipid free formulation could explain the higher potency associated with this formulation. BT were independent of formulation and correlated with values reported for effect-site equilibration half-time consistent with a distribution mechanism for pharmacologic hysteresis. PMID- 9743406 TI - Treatment of pulmonary hypertension and hypoxia due to oleic acid induced lung injury with intratracheal prostaglandin E1 during partial liquid ventilation. AB - BACKGROUND: Partial liquid ventilation using perfluorocarbon liquids may be of therapeutic benefit in patients with acute respiratory failure. This study investigated the effects of prostaglandin E1 (PGE1) delivered intratracheally during partial liquid ventilation on lung function and pulmonary circulation in rabbits with acute respiratory distress syndrome. METHODS: Lung injury was induced by intravenous oleic acid in adult Japanese white rabbits, 1 h after which they were divided into four groups of 10 animals. Group 1 received mechanical ventilation alone, group 2 received aerosolized PGE1 (5 microg followed by 0.1 microg x kg(-1) x min(-1)) under mechanical ventilation combined with 5 cm H2O positive end-expiratory pressure, and groups 3 and 4 received partial liquid ventilation with 15 ml/kg perflubron. Group 4 received a 5-microg bolus followed by 0.1 microg x kg(-1) x min(-1) PGE1 instilled intratracheally (not by aerosol) in combination with partial liquid ventilation. Measurements were performed at 30-min intervals for 120 min after lung injury. RESULTS: After lung injury, hypoxemia, hypercapnia, acidosis, and pulmonary hypertension developed in all animals and were sustained in groups 1 and 2 throughout the experiment. The partial pressure of oxygen in arterial blood of animals in group 3 improved with initiation of treatment, with statistical significance achieved at the 30 and 60 min time points as compared with controls. Group 4 animals had immediate and sustained increases in the partial pressure of oxygen in arterial blood that were significant compared with all other groups during the experiment. Statistically significant reductions in mean pulmonary artery pressure were seen only in group 4 animals compared with all other groups. CONCLUSIONS: These results suggest that PGE1 delivered intratracheally during partial liquid ventilation may be a useful therapeutic strategy for patients with the acute respiratory distress syndrome. PMID- 9743407 TI - Treatment of normal skeletal muscle with FK506 or rapamycin results in halothane induced muscle contracture. AB - BACKGROUND: FKBP12 is a protein that is closely associated with the ryanodine receptor type 1 of skeletal muscle and modulates Ca2+ release by the channel. The immunosuppressants FK506 and rapamycin both bind to FKBP12 and in turn dissociate the protein from the ryanodine receptor. By treating healthy human skeletal muscle strips with FK506 or rapamycin and then subjecting the strips to the caffeine-halothane contracture test, this study determined that FK506 and rapamycin alter the sensitivity of the muscle strip to halothane, caffeine, or both. METHODS: Skeletal muscle strips from 10 healthy persons were incubated in Krebs medium equilibrated with a 95% oxygen and 5% carbon dioxide mixture, which contained either 12 microM FK506 (n = 8) or 12 microM rapamycin (n = 6), for 15 min at 37 degrees C. The strips were subjected to the caffeine-halothane contracture test for malignant hyperthermia according to the European Malignant Hyperthermia Group protocol. RESULTS: Treatment of normal skeletal muscle strips with FK506 and rapamycin resulted in halothane-induced contractures of 0.44+/ 0.16 g and 0.6+/-0.49 g, respectively, at 2% halothane. CONCLUSIONS: The results obtained show that pre-exposure of healthy skeletal muscle strips to either FK506 or rapamycin is sufficient to give rise to halothane-induced contractures. This is most likely caused by destabilization of Ca2+ release by the ryanodine receptor as a result of the dissociation of FKBP12. This finding suggests that a mutation in FKBP12 or changes in its capacity to bind to the ryanodine receptor could alter the halothane sensitivity of the skeletal muscle ryanodine receptor and thereby predispose the person to malignant hyperthermia. PMID- 9743408 TI - Nefiracetam prevents propofol-induced anterograde and retrograde amnesia in the rodent without compromising quality of anesthesia. AB - BACKGROUND: Propofol is a short-acting intravenous anesthetic agent. However, cognitive function remains depressed for several hours thereafter. We have evaluated the ability of nefiracetam, a novel cognition-enhancing agent, to alleviate propofol-induced amnesia in a rodent model of learning. METHODS: Rats were trained in a one-trial, step-through, light-dark passive avoidance paradigm. Propofol (10 and 75 mg/kg) was administered by the intraperitoneal route at 15 min before training and separately at increasing times in the immediate 0-6 h post-training period (100 and 150 mg/kg). Nefiracetam, 9 mg/kg, was administered by the intraperitoneal route 1 h before training. Animals were tested for recall at the 12 h post-training time, and after their killing, immunocytochemistry was used to determine the increase in hippocampal neuronal polysialylation, an event associated with memory consolidation. Induction and duration of anesthesia induced by propofol was determined using tail pinch and pedal withdrawal reflexes. RESULTS: Propofol-induced anterograde amnesia occurred in a dose dependent manner. Induction of retrograde amnesia required a higher dose of propofol, which anesthetized the animals and was effective only in the immediate 3-h post-training period. In the absence of any evident effect on the onset or duration of anesthesia, nefiracetam prevented both forms of propofol-induced amnesia and preserved the learning-associated changes of neuronal polysialylation state. CONCLUSIONS: The ability of nefiracetam to prevent propofol-induced anterograde and retrograde amnesia is proposed to be indirect and to result from modulation of gene transcription in a manner that initiates a cascade of events involving protein synthesis leading to synaptic growth associated with the formation of the long-term memory trace. PMID- 9743409 TI - Inhibition of the enzymic degradation of suxamethonium and mivacurium increases the onset time of submaximal neuromuscular block. AB - BACKGROUND: The factors that influence the onset time of submaximal (<100%) neuromuscular block are not fully known. The authors hypothesized that differences in the rate of decrease in the plasma concentration result in differences in the rate of equilibration between plasma and biophase and thus in different onset times. If this hypothesis is valid, inhibition of the enzymic degradation of muscle relaxants should increase the onset time of neuromuscular block. METHODS: Twenty pigs received either suxamethonium or mivacurium. Dose finding (70% block) was done for each pig. The enzymic degradation of the muscle relaxant was randomly inhibited by selective inhibition of plasma cholinesterase activity by tetraisopropyl pyrophosphoramide (10 pigs) or was not inhibited (10 pigs). Plasma cholinesterase activities and the mechanomyographic muscle response after peroneal nerve stimulation (0.1 Hz) were measured. RESULTS: Inhibition of plasma cholinesterase activity (by 93% and 89%, respectively) increased the onset time of suxamethonium from a median of 40 s (range, 20-45 s) to 131 s (range, 114 166 s; P = 0.009) and of mivacurium from a median of 52 s (range, 40-59 s) to 105 s (range, 90-125 s; P = 0.009). Inhibition of degradation decreased the effective dose of suxamethonium that resulted in 70% depression of the initial twitch height from 900 microg/kg (range, 400-1,000 microg/kg) to 150 microg/kg (range, 135-150 microg/kg) and of mivacurium from 100 microg/kg (range, 80-150 microg/kg) to 35 microg/kg (range, 20-50 microg/kg). CONCLUSIONS: Inhibition of the enzymic degradation of suxamethonium and mivacurium increases the onset time of submaximal neuromuscular block. Therefore, pharmacokinetics influence the onset time of submaximal neuromuscular block. These results imply that to obtain an ultrashort onset time, muscle relaxants should be developed that not only have a low affinity for the receptor but also rapidly disappear from plasma. PMID- 9743410 TI - Effects of intrathecal NMDA and non-NMDA antagonists on acute thermal nociception and their interaction with morphine. AB - BACKGROUND: N-methyl-D-aspartate (NDMA) antagonists have minimal effects on acute nociception but block facilitated states of processing. In contrast, the alpha amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) antagonists decrease acute noxious responses. Morphine (a mu-opioid agonist) can also decrease acute nociceptive processing. The authors hypothesized that the interaction between morphine and AMPA receptor antagonists would be synergistic, whereas morphine and NMDA antagonists show no such interaction in acute nociception. METHODS: Sprague Dawley rats (weight, 250-300 g) were implanted with chronic lumbar intrathecal catheters and were assigned to receive one of several doses of morphine--ACEA 1021 (NMDA glycine site antagonist), ACEA 2085 (AMPA antagonist), AP-5 (NMDA antagonist), saline or vehicle--and were tested for their effect on the response latency using a 52.5 degrees C hot plate. The combinations of morphine and other agents also were tested. RESULTS: Intrathecal morphine (ED50:2 microg/95% confidence interval, 1-4 microg) and ACEA 2085 (6 ng/2-15 ng), but not AP-5 or ACEA 1021, yielded a dose-dependent increase in the thermal escape latency. A systematic isobolographic analysis was carried out between intrathecal morphine and ACEA 2085 using the ED50 dose ratio of 357:1. A potent synergy was observed with decreased side effects. Morphine dose-response curves were carried out for morphine and fixed doses of ACEA 1021 (12 microg) or AP-5 (10 microg). No synergistic interactions were noted. CONCLUSIONS: Spinal mu-receptor activation and AMPA receptor antagonism showed a synergistic antinociception in response to an acute thermal stimulus. NMDA or NMDA glycine site antagonism had no effect alone nor did they display synergy with morphine. These results suggest an important direction for development of acute pain strategies may focus on the AMPA receptor. PMID- 9743411 TI - Cerebellar nitric oxide is increased during isoflurane anesthesia compared to halothane anesthesia: a microdialysis study in rats. AB - BACKGROUND: This study examined the influences of isoflurane versus halothane anesthesia on basal and agonist-stimulated nitric oxide in the cerebellum of intact rats. Nitric oxide was measured using the hemoglobin-trapping method in an in vivo microdialysis technique. This method uses the stoichiometric reaction of nitric oxide with oxyhemoglobin to produce methemoglobin and nitrate; the change in methemoglobin concentration is measured spectrophotometrically to estimate nitric oxide concentration. METHODS: Male Wistar rats were anesthetized with isoflurane (1.4%) or halothane (1.2%), mechanically ventilated and paralyzed (intravenous pancuronium, 1 mg/kg). Microdialysis probes were implanted into the cerebellum. Bovine oxyhemoglobin dissolved in artificial cerebrospinal fluid was pumped through the probe (2 microl/min) and assayed at 15-min intervals. The glutamatergic agonist, kainic acid (KA, 5 mg/kg, intraarterially), was used to stimulate nitric oxide production. NG-nitro L-arginine methyl ester (L-NAME, 40 mg/kg, intravenously) was used to inhibit nitric oxide synthase. RESULTS: Unstimulated cerebellar nitric oxide concentrations were stable and greater during anesthesia with isoflurane (532+/-31 nM; mean +/- SEM) than with halothane (303+/-23 nM). L-NAME pretreatment reduced nitric oxide concentrations during isoflurane, but not halothane, anesthesia. Infusion of KA increased nitric oxide in both groups; however, the increase in nitric oxide was significantly greater during isoflurane anesthesia. Pretreatment with L-NAME inhibited the response to KA in both groups. CONCLUSIONS: Nitric oxide production in the cerebellum, monitored by microdialysis, was greater during isoflurane anesthesia than during halothane anesthesia. Increased nitric oxide production during isoflurane anesthesia would be expected to impact central neuronal function and cerebral blood flow and vascular resistance. PMID- 9743412 TI - Mechanisms of isoflurane-increased submaximum Ca2+-activated force in rabbit skinned femoral arterial strips. AB - BACKGROUND: Isoflurane enhances contraction in isolated intact arterial rings by a protein kinase C (PKC) activator and also causes contracture in skinned arterial strips. This study investigated the mechanisms of this isoflurane activation of the contractile proteins of skinned strips. METHODS: The skinned strips, mounted on photodiode force transducers, were prepared from rabbit femoral arteries treated with saponin. The strips were activated by 1 microM Ca2+ (buffered with 7 mM EGTA) with or without inhibitors for PKC and calmodulin dependent protein kinase II (CaM kinase II). When force reached steady state, isoflurane was administered and changes in force were observed. Another group of the strips was frozen to assay myosin light chain phosphorylation (MLC-p) using two-dimensional electrophoresis and immunoblotting. Analysis of variance was used to compare the results from test and control groups. Probability values <0.05 were significant. RESULTS: Isoflurane (1-5%) dose dependently increased (24-81%) the Ca2+-activated force. At 1% and 5% isoflurane, MLC-p did not change either as the force increased or reached a new steady state level. However, with 3% isoflurane, MLC-p transiently decreased (29.1% and 17.1% of total MLC for 0% and 3% isoflurane, respectively). The 3% isoflurane-increased force was blocked by 10 microM bisindolymaleidmide, an inhibitor of PKC, but not by 10 microM Go-6976, an inhibitor of Ca2+-dependent PKC, and was enhanced 50% by 0.1 mM KN-62, an inhibitor of CaM kinase II. CONCLUSIONS: Isoflurane increased submaximum Ca2+ activated force in skinned femoral arterial strips by activating Ca2+-independent PKC, possibly epsilon isoezyme. The isoflurane-decreased MLC-p may be caused by activation of CaM kinase II. PMID- 9743413 TI - Dexmedetomidine produces similar alterations in the determinants of left ventricular afterload in conscious dogs before and after the development of pacing-induced cardiomyopathy. AB - BACKGROUND: The authors tested the hypothesis that intravenous dexmedetomidine produces alterations in left ventricular (LV) afterload that are deleterious to cardiac performance in conscious dogs with pacing-induced cardiomyopathy. METHODS: Dogs (n = 8) were fitted with instruments for long-term measurement of LV and aortic blood pressure, aortic blood flow, and subendocardial segment length and received dexmedetomidine (1.25, 2.5, and 5 microg/kg) in a cumulative manner before and after 19+/-3 (mean +/- SEM) days of rapid LV pacing. LV afterload was measured with aortic input impedance [Zin(omega)] and quantified with a three-element Windkessel model. Hemodynamics and Zin(omega)) were assessed under control conditions and 5 and 60 min after administration of each dose. RESULTS: Dexmedetomidine caused early and late decreases in heart rate, the maximum rate of increase of LV pressure, mean aortic blood flow, and stroke volume in dogs before and after pacing. Dexmedetomidine caused similar early increases in total arterial resistance and decreases in total arterial compliance in dogs before and after pacing. Early dexmedetomidine-induced increases in resistance and decreases in compliance caused similar reductions in mean aortic blood flow in cardiomyopathic compared with healthy dogs. Resistance and compliance returned to control values, and characteristic aortic impedance decreased late after dexmedetomidine in healthy dogs. In contrast, resistance remained elevated late after dexmedetomidine in dogs with dilated cardiomyopathy. CONCLUSIONS: Dexmedetomidine causes similar alterations in hemodynamics and LV afterload in conscious dogs with and without pacing-induced cardiomyopathy. PMID- 9743414 TI - Anesthesia for major thermal injury. PMID- 9743415 TI - Cardiac massage: a method rescued from oblivion. PMID- 9743416 TI - Emergent cardiopulmonary bypass in five patients with heparin-induced thrombocytopenia type II employing recombinant hirudin. PMID- 9743417 TI - Severe hypercapnia induced by acute dissecting aortic aneurysm. PMID- 9743418 TI - Early fat embolism after liposuction. PMID- 9743419 TI - Vocal cordal bowing as a cause of long-lasting hoarseness after a few hours of tracheal intubation. PMID- 9743420 TI - There is no golden yardstick. PMID- 9743421 TI - Management of chronic nonmalignant pain. PMID- 9743422 TI - Anesthesia safety, outcomes, and prospective study design. PMID- 9743423 TI - Henry K. Beecher, M.D.: an historical perspective? PMID- 9743424 TI - Effects of atenolol on postoperative myocardial ischemia. PMID- 9743426 TI - Predicting malignant hyperthermia susceptibility. PMID- 9743425 TI - Beta blockade in non-cardiac surgical patients. PMID- 9743427 TI - Is lack of statistical power always evidence of lack of effect? PMID- 9743429 TI - Potential cause for medication administration error. PMID- 9743428 TI - A method for measuring carbon dioxide at the tracheal stoma. PMID- 9743430 TI - Another cause of a prolonged downstroke on the capnograph. PMID- 9743431 TI - Create a party mood for a smooth anesthetic induction of children. PMID- 9743432 TI - Bronchospasm and "reflex right upper lobe atelectasis". PMID- 9743433 TI - Complex regional pain syndrome type II as a complication of subclavian catheter insertion. PMID- 9743434 TI - Use of a guide wire to facilitate tracheal intubation with the Bullard laryngoscope. PMID- 9743435 TI - Air embolism during anesthesia for shoulder arthroscopy. PMID- 9743436 TI - Online Mendelian Inheritance in Man. PMID- 9743437 TI - The calcium channel antagonist debate: recent developments. AB - Recent contributions to the calcium channel antagonist (CCA) debate concern the recommendations in the Sixth Report of the U.S.A. Joint National Committee on Prevention. Detection, Evaluation and Treatment of High Blood Pressure and raise questions about evidence-based medicine. Also, several studies have recently been published. A 4695-patient placebo-controlled trial showed that the CCA nitrendipine significantly reduces cardiovascular complications in older people with systolic hypertension. A 470-patient trial showed a significantly higher rate of myocardial infarction in hypertensive non-insulin-dependent diabetics treated with the CCA nisoldipine than in those treated with the ACE inhibitor enalapril. Non-experimental data (on 84 093 person-years in total) comparing antihypertensive agents in hypertensive women showed mostly non-significant covariate-adjusted total and cardiovascular mortality differences that are difficult to interpret because of potential residual confounding. One report suggested a relationship between CCAs and suicide. A case-control study (9513 cases, 6492 controls) showed that CCAs are unlikely to cause cancer. The current recommendation remains to start treatment of hypertension with diuretics and beta blockers, but recent studies support the use of sustained release CCAs in cases in which blood pressure cannot be controlled with other agents. PMID- 9743438 TI - Sustained-release calcium channel antagonists in cardiovascular disease: pharmacology and current therapeutic use. AB - Calcium channel antagonists are a heterogeneous group of drugs with differing cardiovascular effects, and are effective in the treatment of hypertension and angina pectoris. A number of these agents are available in a sustained-release formulation. These formulations produce a gradual and sustained drop in peripheral vascular resistance, thereby avoiding reflex sympathetic stimulation, and thus may avoid the deleterious effects that have been reported with short acting preparations. The results of a number of trials with sustained-release calcium channel antagonists have been reported recently, and the results are promising. Currently they can be recommended for use in patients with hypertension or stable angina which cannot be controlled with other agents. They have been shown to be at least safe when used in patients with left ventricular systolic dysfunction, and may be useful in certain subsets of patients with heart failure. The empirical use of calcium channel antagonists in unstable angina and acute myocardial infarction is not supported by the currently available data. Several large trials with sustained-release formulations are ongoing, which may alter treatment recommendations in the future. PMID- 9743439 TI - Clinical outcome studies of anti-anginal drug therapy for patients with stable coronary disease: an indication for clinical trials. AB - Despite recent advances in prevention and treatment, the number of individuals who suffer from chronic symptomatic coronary artery disease is rising. Past experience with clinical trials in cardiovascular disease has shown that the results are often unanticipated and at odds with the scientific rationale that existed beforehand. Also, important lessons have been learned concerning sample size and the choice of end-points. One of these has been the unreliability of surrogates (such as blood pressure, exercise capacity and number of anginal attacks) in predicting the effect of treatment on morbidity and mortality. The current drug therapy for staple angina in patients without a history of myocardial infarction has not been tested in large-scale clinical trials. No data exist for organic nitrates. The rationale for using beta-blockers comes from trials in patients who had a history of myocardial infarction. Recent experience with sustained release calcium channel antagonists suggests that they appear to be safe in patients with stable angina. Their effect on overall clinical outcome remains to be established, however. It is for this reason that trials such as ACTION are both justifiable and needed. PMID- 9743440 TI - Design and current status of ACTION: A Coronary disease Trial Investigating Outcome with Nifedipine GITS. Gastro-Intestinal Therapeutic System. AB - AIMS: To present the design of ACTION (A Coronary disease Trial Investigating Outcome with Nifedipine GITS), an ongoing multicentre clinical outcome trial with nifedipine GITS (Gastro-Intestinal Therapeutic System) in patients with stable angina pectoris. METHODS: At least 6000 patients with optimally treated stable angina without depressed left ventricular function are randomized in equal proportions to either nifedipine GITS or matching placebo (starting dose 30 mg, maintenance dose 60 mg once daily). Patients are followed for at least four years. The primary end-point, to be analyzed by assigned treatment, includes all cause mortality, acute myocardial infarction, emergency coronary angiography for refractory angina, overt heart failure, debilitating stroke and peripheral revascularization. For this end-point, the trial has a power of 95% to detect a relative risk reduction of 18% at the 5%, level of significance, and is large enough to exclude an excess mortality caused by nifedipine GITS of 3.1 deaths per 1000 years of treatment or greater. The pre-specified early termination rule is more conservative in the case of a beneficial effect than in the case of an adverse effect of nifedipine GITS. The first patient was randomized on 29 November, 1996. By the end of April 1998, about 5200 patients had been started on study medication. CONCLUSIONS: Results will be available in the autumn of 2003. PMID- 9743441 TI - H-NS controls the transcription of three promoters of Vibrio fischeri lux cloned in Escherichia coli. AB - We have recently proposed that the expression of V. fischeri right lux operon is controlled by two promoters; the first one located upstream of the luxl gene, while the second one seems to be located upstream of the luxC gene. The transcription from both promoters is negatively controlled by H-NS protein. Escherichia coli MC4100 rpoS hns mutant that carried the V. fischeri lux system with a deletion in either the luxl or luxR gene showed a constitutive mode and more than 10,000-fold higher luminescence than the control cells. The present study shows that neither luxR nor luxl are required for the transcription of the luxCDABE genes in an H-NS deficient strain of E. coli. The MC4100 rpoS hns mutant harbouring the luxCDABE-carrying plasmid showed constitutive mode and 70,000-fold higher luminescence than the wild-type cells. The question whether both the left and the right operons of V. fischeri lux system are controlled by H-NS was addressed with the aid of plasmids harbouring the lacZ gene fused with luxR or luxl. In MC4100 hns rpoS background, luxR and luxl genes were very early and actively transcribed, as judged by the strong beta-galactosidase activity that was developed at early stage of growth. The beta-galactosidase activity in the wild-type cells was 20-40 times lower and occurred mainly during the second half of the growth cycle. It thus appears that H-NS inhibits the transcription of three promoters of the lux system of V. fischeri; the left operon that codes for LuxR protein and two promoters located upstream and downstream to luxl gene. PMID- 9743442 TI - Prospects for chemiluminescent and bioluminescent immunoassay and nucleic acid assays in food testing and the pharmaceutical industry. AB - The sensitivity, speed and convenience of chemiluminescent (CL) and bioluminescent (BL) immunoassays and probe assays have led to a diverse range of applications for these technologies, mainly in the clinical laboratory. These methods are now being explored by the food and pharmaceutical industries. Demanding detection limits and the complexity of sample preparation for food and pharmaceutical analyses present daunting challenges for the analyst. Immunoassay and nucleic acid amplification technologies have been applied to food testing, but these have mostly favoured non-luminescent endpoints. Food assays with CL or BL endpoints are now emerging, e.g., Clostridium botulinum type A detection using a CL immunosorbent assay; Salmonella and Zygosaccharomyces detection using a combination of PCR and CL detection. The analytical challenges posed by the pharmaceutical industry include testing for contaminants in raw materials and drug products, and drug discovery. The sensitivity and rapid signal acquisition characteristics of CL and BL are advantageous for the high throughput, massively parallel testing of micro-sized samples demanded in drug discovery. Current progress and the prospects for CL and BL immunoassay and nucleic acid technologies in this and other pharmaceutical and food applications is reviewed. PMID- 9743444 TI - Polyorganophosphazene microspheres for drug release: polymer synthesis, microsphere preparation, in vitro and in vivo naproxen release. AB - Microsphere preparation for naproxen slow release was investigated using two newly prepared biodegradable polyorganophosphazenes, derivatized at the phosphorus atoms with phenylalanine ethyl ester and imidazole at molar ratios of 71/29 and 80/20. The polymers were prepared by substitution of the chloride atoms of polydichlorophosphazene with a phenylalanine ethyl ester-imidazole mixture followed, after 7 or 48 h reaction, by the addition of excess imidazole. Three methods of microsphere preparation have been considered: spray-drying, emulsion/solvent evaporation and emulsion/solvent evaporation-extraction. Microparticles obtained by spray-drying were found to possess a narrow distribution size with a mean diameter of 2-5 microm. Their internal structure consisted of a porous or empty core depending upon the solvent used for the preparation. Furthermore the microspheres prepared with this technique rapidly released the entrapped naproxen independently of the used polymer, the drug loading or the preparation process. On the other hand microspheres prepared by solvent evaporation or solvent evaporation-extraction showed a distribution size ranging between 10 and 100 microm. By the appropriate choice of pH and solvent composition of the external phase, naproxen could be entrapped, in these microspheres, with a yield higher of 80%. The polymer composition dictates the in vitro release rate of naproxen from the particles, which was faster when the microspheres were prepared with the polymer at higher imidazole content. In vivo experiments, carried out by subcutaneous implantation in rats of microspheres prepared by solvent evaporation, demonstrated that a constant level of naproxen in plasma could be maintained up to 400 h at a suitable concentration for antinflammatory activity. PMID- 9743443 TI - Chemiluminescent determination of esterases in monocytes. AB - Esterase from monocytes promotes the hydrolysis of 2-methyl-1-propenylbenzoate (MPB) yielding 2-methyl-1-propenol, which is oxidized by horseradish peroxidase/H2O2 producing triplet acetone. The chemiluminescence of this reaction can be enhanced by the addition of 9,10-dibromoanthracene-2-sulphonate. The non specific esterase present in monocytes is responsible for MPB hydrolysis, since (a) the chemiluminescence of the reaction was inhibited by fluoride, and (b) cells that do not contain a significant amount of non-specific esterases, e.g. lymphocytes and neutrophils, did not trigger light emission. The analytical application of this reaction is considered. PMID- 9743445 TI - Pharmacokinetic characteristics and therapeutic effects of mitomycin C-dextran conjugates after intratumoural injection. AB - The pharmacokinetics and therapeutic effects of macromolecular prodrugs of mitomycin C (MMC), MMC-dextran conjugates (MMC-D) were studied after intratumoural injection in rats bearing Walker 256 carcinosarcoma. As the first step, the intratumoural disposition characteristics of these drugs were delineated in perfusion experiments employing a tissue-isolated tumour preparation. While MMC immediately disappeared from the tumour preparation following direct intratumoural injection, cationic and anionic MMC-D were retained in the tumour longer, demonstrating that the intratumoural clearance of MMC can be greatly retarded by dextran conjugation. The effect was more pronounced in the case of the cationic conjugate. Venous outflow data in the perfusion experiments were analyzed based on a compartment model in which the tumour tissue was assumed to consist of two compartments, one well- and the other poorly-perfused. The pharmacokinetic analysis revealed that macromolecular conjugation reduced elimination of MMC from the poorly-perfused region rather than well-perfused region. Simulation of conjugated and free MMC levels in the tissue using the calculated parameters clearly showed that intratumoural injection of MMC-D, especially the cationic form, can maintain a certain level of active free MMC in the tissue for a much longer time period. The long retention of cationic MMC-D in tumour after intratumoural injection was also confirmed by an in vivo pharmacokinetic study and whole body autoradiography in rats bearing subcutaneous Walker 256 carcinosarcoma. In addition, superior antitumour activity of cationic MMC-D was observed against subcutaneous tumours after intratumoural injection. Together with the finding that MMC is selectively toxic to hypoxic tumour cells at low concentrations, these pharmacokinetic studies strongly support the therapeutic efficacy of the macromolecular prodrugs. PMID- 9743446 TI - Targeting of human and mouse T-lymphocytes by monoclonal antibody-HPMA copolymer doxorubicin conjugates directed against different T-cell surface antigens. AB - Binding of HPMA copolymer-conjugated doxorubicin targeted with monoclonal antibodies directed against various T-cell surface receptors, i.e. Thy1.2 (CDw90), I-A (MHC class II. glycoprotein), L3T4 (CD4), IL-2R (CD25) and CD3, is considerably increased in Con A stimulated T-lymphocytes. FACS analysis showed that the binding is most intensive with anti-Thy1.2 and anti-L3T4 targeted derivatives and it is proportional to the antiproliferative effect of the antibody-targeted drug. No binding and no antiproliferative capacity was observed after in vitro incubation of mouse T-cells with a nonspecific mouse IgG-HPMA-DOX conjugate. [3H]-TdR incorporation was inhibited considerably more in Con A stimulated T-cell culture and in EL4 mouse T-cell lymphoma as compared with the culture of nonactivated T-lymphocytes. This proves that intensively proliferating cells are more susceptible to the inhibitory action of an antibody-targeted drug. The cytotoxic efficacy of HPMA copolymer with GlyPheLeuGly or GlyLeuPheGly side chains to which the drug is conjugated was superior to HPMA copolymer with GlyPheGly or GlyLeuGly side-chains. However, there is no direct correlation between the rate of in vitro drug release and the in vitro cytotoxicity of the respective conjugates. This suggests that the rate of drug release from the conjugate is only one factor responsible for the pharmacological efficacy of the preparation. Furthermore, we detected substantial and prolonged inhibition of proliferation of Con A activated T-cells only if doxorubicin was injected in vivo in the form of an anti-Thy1.2-targeted conjugate. PMID- 9743447 TI - Phase coexistence in cholesterol-fatty acid mixtures and the effect of the penetration enhancer Azone. AB - Small and wide-angle X-ray diffraction was used to study the phase behaviour of cholesterol-fatty acid mixtures in an attempt to understand lipid interaction occurring in the stratum corneum, the outermost layer of skin. The effect of the penetration enhancer Azone was investigated as well. It was found that equimolar mixtures of cholesterol, palmitic acid and oleic acid (with the acids neutralised to 41 mol%) in 25% (wt/wt) water typically showed three phases at room temperature, two crystalline and one gel phase. The crystalline phases consisted mainly of palmitic acid:soap and cholesterol, respectively. The water present was unevenly distributed and was associated with the gel phase. Both cholesterol and palmitic acid seemed to be depleted from their crystalline phases by Azone. The electrostatic effects on titration of fatty acids in lamellar aggregates were calculated in view of the present results, and the effects of phase separation were discussed. PMID- 9743448 TI - Effect of geometrical cement size on in vitro and in vivo indomethacin release from self-setting apatite cement. AB - The relationship between in vitro and in vivo indomethacin (IMC) release from a self-setting bioactive apatite cement and cement size were investigated. Differently sized apatite cements (total weight, 500 mg); either 64 of the small size (2 mm diameter x 2 mm thickness), sixteen of the medium size (4 mm x 2 mm) or one of the large size (15 mm x 2 mm) were obtained from cement bulk powder containing tetracalcium phosphate, dicalcium phosphate dihydrate and hydroxyapatite. In vitro IMC release from the 1, 2 and 5% drug-loaded apatite cement systems in simulated body fluid (SBF) (pH 7.25) at 37 degrees C increased with increasing concentrations of IMC and with decreasing geometrical size of the cement. The plots of in vitro IMC release per unit area against the square root of time increased with increasing IMC concentrations, but not with decreasing geometrical size of the cement. After subcutaneous (s.c.) implantation of differently sized 1% IMC-loaded cements in male Wistar rats, the plasma IMC concentration and the area under the curve increased with decreasing cement diameter. The in vivo IMC release profiles of the cement were deconvoluted from the plasma IMC profiles after s.c. administration of IMC solution. The plots of in vivo IMC release per unit area against the square root of time suggested that the initial release from all 1% drug-loaded cements was very rapid, slowed after one day, but continued for over two weeks. The relationship between the in vitro release in SBF and the in vivo release in rats of IMC-loaded cements was linear. PMID- 9743449 TI - Azocross-linked poly(acrylic acid) for colonic delivery and adhesion specificity: synthesis and characterisation. AB - A new synthetic pathway to 4,4'-divinylazobenzene is presented together with a procedure for the copolymerisation of this compound with acrylic acid. The chemical structure of the synthesised series of copolymers is examined in the light of infrared spectroscopy results and nuclear magnetic resonance data. The thermal properties of the materials are assessed using a combination of thermal analysis techniques and their swelling behaviour is evaluated at physiologically relevant buffers designed to mimic the gastrointestinal environment. PMID- 9743450 TI - Drug absorption from nifedipine hydrophilic matrix extended-release (ER) tablet comparison with an osmotic pump tablet and effect of food. AB - The aim of the present study was to compare the bioavailability of nifedipine when administered as a hydrophilic matrix tablet (ER) and a push-pull osmotic pump tablet (XL) administrated after fasting, and to evaluate the effect of food for the hydrophilic matrix tablet. For this purpose, three separate studies were performed on healthy volunteers (n = 58) including gammascintigraphic monitoring of tablet erosion and localisation in the gastrointestinal tract for ER in one study. Both ER and XL provided almost constant drug delivery over 24 h, after administration under fasting conditions, and bioequivalence was obtained according to 90% confidence intervals of the difference between formulations within 80-125% for Cmax and AUC. Food significantly increased AUC for ER but no significant difference was obtained between ER and XL with food with respect to extent of bioavailability. The rate of absorption was increased to a higher degree for ER than for XL, as indicated by a Cmax which was almost twice as high for ER compared with XL. This finding was shown to be related to an increased tablet-erosion rate for ER, leading to more rapid drug release. PMID- 9743451 TI - The control of protein release from poly(DL-lactide co-glycolide) microparticles by variation of the external aqueous phase surfactant in the water-in oil-in water method. AB - Poly(DL-lactide co-glycolide) microparticles below 5 microm in size and containing ovalbumin (OVA), were prepared using the water-in oil-in water (w/o/w) technique with either polyvinyl alcohol (PVA) or polyvinylpyrrolidone (PVP) as stabilisers in the external aqueous phase. PVP-stabilised microparticles exhibited higher protein loading (8.2%, w/w relative to 4.0% for PVA stabilised microparticles) and increased core loading (encapsulation) of protein (70% vs. 30% for the PVA system). The use of PVP instead of PVA to prepare microparticles also resulted in reduction in the initial burst release of OVA, together with sustained protein release over 28 days and an increase in the protein delivery capacity from 35 to 45 microg/mg particles. The changes in protein loading and delivery characteristics are considered to arise in part from an increase in the viscosity of the droplets of polymer solution, constituting the primary water-in oil emulsion, by diffusion of PVP from the external aqueous phase. Variation of the external aqueous phase surfactant provides a promising approach for improving the loading of therapeutic proteins and vaccine antigens within biodegradable microparticles and for modulating their release pattern. PMID- 9743452 TI - Radiation and thyroid cancer: what more can be learned? PMID- 9743453 TI - Radiation-associated thyroid cancer--facts and fiction. AB - Thyroid cancer is a rare and complex disease and the thyroid contains various cell types from which specific diseases arise. The malignancies range from indolent to extremely aggressive. Several risk factors for thyroid cancers have been suggested, but only ionizing radiation has been found to have a causative effect. With the exception of tobacco, ionizing radiation is probably the most studied carcinogen there is. It is not yet possible to determine whether or not a specific cancer was induced by radiation and epidemiological studies therefore provide the primary data on cancer risk in man after exposure to ionizing radiation. The current epidemiological knowledge on radiation-associated thyroid cancer is reviewed, focusing on the sharply increased risks recently found among children in the Chernobyl area. PMID- 9743454 TI - Interferon and malignant disease--how does it work and why doesn't it always? AB - Since their first use at the clinic almost 30 years ago. interferons (IFNS) have become an accepted therapy in a range of malignancies. Although IFN All induce remissions in some patients, they are of no benefit, or at best, lead only to minor improvements in the great majority of patients. This review considers possible mechanisms underlying the antitumour effects of IFN, and discusses possible reasons for resistance to IFN therapy in patients with malignant disease. PMID- 9743455 TI - A critical review of papers from clinical cancer research. AB - A review of 75 original articles from clinical cancer research in Norway is presented. Articles published in 1993 and with at least one Norwegian author were included in the review. Sixty papers were observational, whereas 15 were experimental. Of the observational studies 44 were retrospective. Most of the papers concerned prognostic factors. Prior hypotheses were explicitly defined in 16 papers only, and less than half of the articles described inclusion and exclusion criteria. Sample size calculations were performed in four papers only. The choice of statistical method was considered to be suitable in 22 of the 58 articles presenting statistical inferences. Problems related to multiple significance testing were rarely addressed, although the median number of p values reported was as high as 8. Confidence intervals for main findings were presented in 14 papers. For proper planning of studies, as well as for analysis and interpretation of study results, statistical advice is indeed required. PMID- 9743456 TI - Histamine and cytokine therapy. AB - Interleukin-2 (IL-2) and interferon-alpha (IFN-alpha) are potent activators of natural killer (NK) cells and other anti-tumor effector cells, but the results obtained in clinical trials with these cytokines have proved disappointing in many forms of cancer. It may be that IL-2 and IFN-alpha are often not sufficiently effective because intratumoral monocytes/macrophages (MO) inhibit the cytokine-induced activation of cytotoxic effector lymphocytes such as NK cells at the site of tumor growth. An essential part of this inhibitory signal is conveyed by MO-derived reactive oxygen species (ROS), which potently inhibit NK cell-related functions, including the constitutive and cytokine-induced cytotoxicity against tumor cells. Histamine, a biogenic amine, inhibits ROS formation in MO; thereby, histamine synergizes with IL-2 and with IFN-alpha to induce killing of NK-cell-sensitive human tumor cells in vitro. Furthermore, treatment of tumor-bearing mice with histamine potentiates cytokine-induced killing of NK-cell-sensitive murine tumor cells in vivo. In ongoing clinical trials, histamine has been added to IL-2 or IFN-alpha in immunotherapy of human neoplastic disease. The results of two pilot trials in metastatic melanoma suggest that the addition of histamine to IL-2/IFN-alpha prolongs survival time and induces regression of tumors, such as liver melanoma, which are considered refractory to immunotherapy with IL-2 or IFN-alpha. In acute myelogenous leukemia (AML), histamine and IL-2 have been given in order to protect patients in remission against relapse of leukemic disease. The potential benefit of histamine therapy in melanoma and AML will be evaluated in randomized trials. PMID- 9743457 TI - Cisplatin-induced inhibition of p34cdc2 is abolished by 5-fluorouracil. AB - Clinical (Dimery and Hong, J Nat Cancer Inst 1993; 85: 95- 111) and experimental studies (Scanlon et al., Proc Natl Acad Sci USA 1986; 83: 8923-5; Lewin et al., In Vivo 1990; 4: 277-82) have indicated an increased cytotoxic effect, when cisplatin (CDDP) is combined with 5-fluorouracil (5-FU). Addition of 5-FU abolishes the G2-arrest induced by CDDP (Lewin et al., In Vivo 1990; 4: 277-82; Nylen et al., Acta Oncol 1996; 35: 229 35). The mechanism for the synergy is unclear. Activation of p34cdc2 is necessary for progression from G2 to mitosis (Lewin et al., Anti-Cancer Drugs 1995; 6: 465-70). The aim was to study p34cdc2, cdc25C and weel after treatment of mammalian tumour cells in vivo with CDDP as single agent or in combination with 5-FU. CDDP prevented activation of p34cdc2 by keeping cdc25C inactive and weel active. Addition of 5-FU to CDDP decreased the expression of weel and promoted cdc25C-activation. p34cdc2 was dephosphorylated by cdc25C and activated. Alterations in activity of cdc25C and weel after drug combination were due to changes in the protein amount, rather than to changes in the phosphorylation degree. PMID- 9743458 TI - Influence of droloxifene on metastatic breast cancer as first-line endocrine treatment. AB - The effect of droloxifene (3-hydroxytamoxifen) given as first-line endocrine treatment was evaluated in 39 postmenopausal women with advanced receptor positive or receptor-unknown breast cancer. The patients had not received any previous anticancer therapy apart from adjuvant treatment. The overall response rate (CR + PR) was 51% (8% CR, 43% PR), 95% confidence interval+/-15.7%. Median time to progression (all patients) was 8 months, the median time to response 2 months, while the median duration of response was 10 months. The drug was well tolerated with no major side effects recorded; 16% of the patients experienced hot flushes. The response to droloxifene recorded in the present study is in accordance with the response rates to tamoxifen as first-line treatment in identical groups of patients. PMID- 9743459 TI - Experimental studies on the possible influence of invasive oxygen measurements on tumour radiosensitivity. AB - The effects of tissue damage associated with invasive pO2 measurements on radiation sensitivity were investigated using a xenografted squamous cell carcinoma model. For the tumour cure experiments, single dose irradiations were given following different regimens of polarographic pO2 measurements associated with different degrees of mechanical tissue damage. With a dose of 32 Gy, 57% of animals were cured. Following 3 tracks of needle measurements, 73% of tumours were locally controlled, and 75% were cured after 8 needle tracks. The polarographic measurements gave virtually identical oxygenation data for recurrent or cured tumours (both median pO2 1.0 mmHg), respectively. There was thus no evidence of decreased radiosensitivity associated with tissue damage after invasive pO2 measurements. The pre-therapeutic oxygenation status gave no evidence for a prediction of radiation response on an individual basis. PMID- 9743461 TI - Bilateral ovarian metastases from renal clear cell carcinoma. PMID- 9743460 TI - Ewing's sarcoma treatment in Scandinavia 1984-1990--ten-year results of the Scandinavian Sarcoma Group Protocol SSGIV. AB - A report on the long-term follow up of the first cooperative Scandinavian Sarcoma Group study in Ewing's sarcoma of bone is presented. Fifty-two previously untreated patients entered the study between 1984 and 1990. Half of the tumors were located in the extremities and one quarter in the pelvis. The combined modality treatment consisted of 5 cycles of chemotherapy--including vincristine, methotrexate, doxorubicin, cyclophosphamide, bleomycin and dactinomycin. The first two cycles were followed by local resection or amputation in 35 patients and by radiotherapy alone in 17 patients. When surgery was not performed, was incomplete or yielded poor margins radiotherapy was given at a dose of 40-60 Gy. Local tumor relapses developed in 10 patients and in all but one patient were accompanied by metastatic disease. Five patients had metastasis at diagnosis and distant metastases developed after primary treatment in 27 patients after a median of 14 months. The median follow-up time for the 20 surviving patients is 10 years. At 5 years the tumor-related survival was 46% and the metastasis-free survival 43%. Late tumor relapses occurred in 4 patients, which reduced the 10 year tumor related survival to 41% and the metastasis-free survival to 38%. Histopathological tumour response correlated with survival with 5-year metastasis free survival rates of 73% for the good responders and 35% for the poor responders. PMID- 9743463 TI - Vitravene--another piece in the mosaic. PMID- 9743462 TI - Primary malignant melanoma arising in an ovarian cystic teratoma. PMID- 9743464 TI - Limited use of cationic liposomes as tools to enhance the antiherpetic activities of oligonucleotides in vero cells infected with herpes simplex virus type 1. AB - We used commercially available cationic liposomes, lipofectin, DOTAP, and transfectam, to enhance the antiherpetic activities of phosphodiester oligonucleotides (D-oligos) or phosphorothioate oligonucleotides (S-oligos) targeted against immediate-early pre-mRNA4/5 of herpes simplex virus type 1 (HSV 1). With a 5-fold excess of S-oligos/D-oligos, formation of complexes with some of the S-oligos/D-oligos and the cationic liposomes could be visualized on agarose gel. A >5-fold excess of cationic liposomes enhanced the antiherpetic activities of Doligos, whereas there was not enhancement of the antiherpetic activities of S-oligos. As nuclear localization of D-oligos in the presence of cationic liposomes was not clear, we could not clarify the relation between antiherpetic activities of D-oligos and nuclear distribution of oligos. Subcellular distribution of S-oligos in the presence of lipofectin or DOTAP showed nuclear localization by confocal laser scanning microscopy. Transfectam had no effect on the nuclear distribution of S-oligos. These data showed that cationic liposomes would not be appropriate carriers to enhance the antiherpetic activities of S-oligos. Also, distribution of S-oligos into the nucleus does not necessarily enhance their biologic activity. Questions remain about the effectiveness of cationic liposomes in the enhancement of the antivirus activity of S-oligos. PMID- 9743465 TI - Oligonucleotide inhibitors of P-selectin-dependent neutrophil-platelet adhesion. AB - P-selectin, an inducible cell adhesion molecule, mediates rolling of neutrophils on activated vascular endothelium. Because rolling is an early event of the inflammatory response, therapeutic applications of selectin antagonists have been of broad interest. There are, however, no truly satisfactory therapeutic candidates among known inhibitors. Consequently, we have used Systematic Evolution of Ligands by Exponential Enrichment (SELEX) technology, a process based on oligonucleotide combinatorial chemistry and in vitro selection, to develop aptamer antagonists of P-selectin. Equilibrium dissociation constants for aptamer/P-selectin binding range from 16 to 710 pM, a 10(5)-10(6)-fold improvement compared with the minimal carbohydrate ligand, sialyl Lewis X (sLeX). Aptamer binding is divalent cation dependent and, unlike sLeX, is specific for P selectin. The selectivity for human P-selectin relative to human E-selectin or human L-selectin is 10(4)-10(5). In vitro, aptamers bind with subnanomolar affinities to P-selectin expressed on thrombin-activated platelets, inhibit the binding of P-selectin-IgG chimera to sLeX and to neutrophils, and block the binding activated platelets to neutrophils in flow cytometry and in hydrodynamic assays. Extrapolating from their in vitro characteristics, these novel P-selectin specific antagonists may be suitable candidates for therapeutic development. PMID- 9743466 TI - The influence of target protein half-life on the effectiveness of antisense oligonucleotide analog-mediated biologic responses. AB - During the course of a study aimed at improving antisense oligodeoxynucleotide mediated ex vivo bone marrow purging of patients suffering from chronic myeloid leukemia (CML), the properties of a number of antisense structures intended to reduce the expression of c-myc, mutant p53, and bcr-abl mRNAs and proteins were examined. The majority of the antisense oligodeoxynucleotides were designed to be capable of directing ribonuclease H (RNase H) cleavage of their target mRNAs. Streptolysin O (SLO) reversible permeabilization was used to deliver the oligodeoxynucleotides into the CML line KYO-1. We found that the efficiency and specificity of antisense oligonucleotide-induced reductions of target protein expression depended on target protein half-life, the oligonucleotide structure, and the specific sequence within the target mRNA. Transient reductions of c-myc mRNA and protein were achieved with a chimeric methylphosphonate-phosphodiester oligodeoxynucleotide antisense to the initiation codon, but cell proliferation was unaffected. In contrast, a chimeric oligodeoxynucleotide of similar structure targeted to an alternative site in the coding region of c-myc mRNA reduced target mRNA and protein levels for over 24 hours and halted cell proliferation. Chimeric methylphosphonate-phosphodiester oligodeoxynucleotide antisense to a point mutation in KYO-1 p53 mRNA efficiently reduced target mRNA expression, but only small, transient reductions in p53 protein expression were observed. However, a chimeric methylphosphonate-phosphorothioate oligodeoxynucleotide targeted to the same site reduced p53 protein to 30% of control levels over a 48-hour period. BCR ABL protein expression was unaffected by chimeric oligodeoxynucleotides targeted to the breakpoint in bcr-abl mRNA, even when mRNA levels at early times were substantially reduced. PMID- 9743467 TI - Effect of antisense oligodeoxynucleotides directed to individual calmodulin gene transcripts on the proliferation and differentiation of PC12 cells. AB - Calmodulin (CaM) is encoded by three different genes that collectively give rise to five transcripts. In the present study, we used antisense oligodeoxynucleotides targeted to unique sequences in the transcripts from the individual CaM genes to selectively block the expression of the different genes and to investigate the roles these individual genes play in the proliferation and nerve growth factor (NGF)-induced differentiation of PC12 cells. Culturing PC12 cells in the presence of oligodeoxynucleotide antisense to the transcripts from CaM genes I and II caused a significant decrease in the proliferation and a significant delay in the NGF-induced differentiation of PC12 cells when compared with untreated cells and with cells treated with the corresponding randomized oligodeoxynucleotides. However, an oligodeoxynucleotide antisense to CaM gene III did not significantly alter the proliferation or the NGF-induced differentiation of PC12 cells. The inhibition of cell proliferation could be reversed by washing out the antisense oligodeoxynucleotides. The levels of CaM in cells treated with oligodeoxynucleotides antisense to CaM genes I or II were reduced 52% or 63%, respectively, of the levels found in the control cells. However, the levels of CaM were not significantly reduced in PC12 cells treated with CaM gene III antisense oligodeoxynucleotide. None of the randomized oligodeoxynucleotides had any effect on the levels of CaM in PC12 cells. The reduced levels of CaM in PC12 cells treated with an oligodeoxynucleotide antisense to CaM gene I were accompanied by a reduction in the levels of the CaM gene I mRNAs, supporting a true antisense mechanism of action for these oligodeoxynucleotides. These results suggest that altering the level of CaM by using antisense oligodeoxynucleotides targeted to the dominant CaM transcripts in a particular cell type will specifically inhibit their proliferation and, in the case of neuronal cells, alter the course of their differentiation. PMID- 9743468 TI - Synthetic 2'-O-methyl-modified hammerhead ribozymes targeted to the RNA component of telomerase as sequence-specific inhibitors of telomerase activity. AB - Telomerase is a ribonucleoprotein that synthesizes tandem arrays of the hexameric DNA sequence TTAGGG at chromosome termini using its RNA component as a template. As most normal cells lack telomerase activity, a progressive shortening of chromosome length occurs with each cell division because of incomplete DNA replication. Cell senescence ensues when a critical telomere length is reached, but importantly, senescence bypass and life span extension occur in normal cells transfected with functional telomerase activity. Almost 90% of all tumors express telomerase activity, implying that telomerase is an important determinant in tumor progression and cell immortalization. However, the exact role and regulation of the individual components of the telomerase complex are not fully understood and would benefit from the availability of specific inhibitors. In this study, we investigated the potential use of chemically stabilized, catalytic RNA molecules (hammerhead ribozymes) to inhibit telomerase activity by cleaving the RNA component in a sequence-specific manner. Catalytically competent (active) hammerhead ribozymes containing 2'-O-methyl ribonucleotides for enhanced biologic stability and designed to be complementary to the RNA component of human telomerase exhibited dose-dependent inhibition of telomerase activity in human glioma U87-MG cell lysates with an IC50 of around 0.4 microM. Catalytically incompetent (inactive) ribozymes or mismatched ribozymes with reduced hybridization capability to telomerase RNA did not inhibit telomerase activity, as detected by a PCR-based telomeric repeat amplification protocol (TRAP) assay. In vitro cleavage reactions using short substrates and RT-PCR analyses of the full-length RNA substrate in U87-MG cell lysates confirmed a sequence-specific catalytic cleavage of the targeted RNA component of telomerase. Exogenously administrable, synthetic ribozymes may have important uses in further understanding the role and regulation of this ribonucleoprotein in normal and diseased tissues as well as in the potential therapy of telomerase-positive tumors. PMID- 9743469 TI - Specific inhibition of interleukin-10 production in murine macrophage-like cells by phosphorothioate antisense oligonucleotides. AB - The effects of phosphorothioate antisense oligonucleotides (AS-S-oligos) directed against murine interleukin-10 (IL-10) mRNA on IL-10 production in RAW264.7 cells, a murine macrophage-like cell line, when stimulated by lipopolysaccharide (LPS) were examined. Of the six AS-S-oligos used, AS-S-oligos directed against the 3' untranslated region (3'-UTR) of IL-10 mRNA (AS6-S-oligo) showed the strongest inhibitory effect on IL-10 production, and this inhibition was dose and time dependent. Reverse transcription-polymerase chain reaction (RT-PCR) revealed that the antisense effect originated from a specific reduction of target IL-10 mRNA by hybridization with AS6-S-oligo. In addition, AS6-S-oligo did not affect tumor necrosis factor-alpha (TNF-alpha) production in cells stimulated by LPS, and S oligos with control sequences did not affect IL-10 production. These findings suggested that AS6-S-oligo most powerfully inhibited IL-10 production in macrophages by an antisense mechanism. PMID- 9743470 TI - Growth inhibition of chronic myelogenous leukemia cells by ODN-1, an aptameric inhibitor of p210bcr-abl tyrosine kinase activity. AB - p210bcr-abl-Related tyrosine kinase activity has been shown to cause chronic myelogenous leukemia (CML), a disease of bone marrow stem cells. Having previously demonstrated that the aptameric oligonucleotide, ODN-1, could inhibit p210bcr-abl kinase activity, the current study sought to determine if ODN-1 could selectively inhibit the growth of CML cells relative to that of normal bone marrow. ODN-1, when introduced by electroporation into peripheral blood mononuclear cells (PBMC) from patients with CML, decreased the number of committed progenitors (CML CFU-GM) by an average of 67%+/-19% (mean+/-SEM, range 28-98%). Treatment of CML PBMC with ODN-1 was also shown to decrease the number of more primitive cobblestone area-forming cells (CAFC) by 35%-87%. In contrast, there was little suppressive effect by the combination of electroporation and ODN 1 on either CFU-GM or CAFC numbers from normal donor bone marrow. These studies suggest that inhibition of p210bcr-abl protein-tyrosine kinase (PTK) activity by ODN-1 is associated with some degree of selective growth inhibition of p210bcr abl-transformed cells. p210bcr-abl kinase inhibitory agents may be useful for the ex vivo purging of bone marrow or peripheral blood progenitor/stem cells in the setting of autologous transplantation for CML. PMID- 9743471 TI - Antisense inhibition of Bax mRNA increases survival of terminally differentiated HL60 cells. AB - Cell sensitivity to programmed cell death is primarily modulated by members of the Bcl-2 family, as the balance of homodimer or heterodimer formation between proapoptotic and antiapoptotic members defines apoptosis susceptibility in the great majority of cellular contexts. It is, therefore, important to clarify if the Bax protein is limiting for activation of the genetic program of programmed cell death or can be complemented by different Bcl-2 family members, such as Bak or Bad. To gain some insight into the role of Bax in the molecular mechanisms of apoptosis of myeloid cells, we inhibited this gene in all-trans-retinoic acid (ATRA)-treated HL60 cells using the methodology of antisense oligodeoxynucleotides (AS-ODN). Our results indicate that Bax inhibition has no effect on the proliferation and differentiation capacity of HL60 cells. Instead, the survival rate of terminally differentiated Bax-inactivated HL60 (Bax(-) HL60) cells is almost three times higher in respect to control cultures, indicating that in mature granulocytes Bax is not efficiently complemented by others members of the Bcl-2 family proteins. PMID- 9743472 TI - Reduction of antigen expression from DNA vaccines by coadministered oligodeoxynucleotides. AB - Bacterial DNA or synthetic oligodeoxynucleotides (ODN) containing unmethylated CpG dinucleotides within the context of certain flanking bases (CpG motifs) have potent stimulatory effects on the vertebrate immune system. CpG ODN with a synthetic nuclease-resistant phosphorothioate backbone (S-ODN) can be used as an adjuvant to augment both humoral and cell-mediated immune responses against a protein antigen. It has also been shown that the presence of CpG motifs in DNA vaccines may be responsible, at least in part, for their efficacy. Here we evaluate the possibility of using CpG ODN as an adjuvant with DNA vaccines to further improve their efficacy. We show that it is not possible to directly mix S ODN with plasmid DNA because this will result in an ODN dose-dependent reduction in gene expression from the plasmid, possibly because of competitive interference at binding sites on the surface of target cells. Although ODN with a phosphorothioate-phosphodiester chimeric backbone (SDS-ODN) do not adversely effect the level of gene expression (except when certain sequences, such as a poly G, are present), this is not useful, as SDS-ODN are apparently also not sufficiently nuclease resistant to exert a strong CpG adjuvant effect. Neither is it possible to augment responses to DNA vaccines by administering the CpG S-ODN at a different time or site than the plasmid DNA. Thus, at least for the present, it appears necessary to clone CpG motifs into DNA vaccine vectors to take advantage of their adjuvant effect. PMID- 9743473 TI - A folate binding protein in ascitic fluid, serum and ovarian tissue of patients with ovarian adenocarcinoma immunoreacts with antibodies against human milk folate binding protein. AB - The presence of a folate binding protein which immunoreacts with antibodies against human milk folate binding protein was demonstrated in ascitic fluids from seven patients with ovarian adenocarcinoma. Ascitic fluids collected from two patients with other malignancies contained non-immunoreactive FBP. Tumor tissue specimens from five patients with ovarian carcinoma contained immunoreactive FBP. By contrast to normal ovaries ovarian carcinoma tissue showed positive immunostaining on immunohistochemistry. Ascitic fluids from two patients with ovarian carcinoma exhibited single distinct bands on SDS-PAGE immunoblotting. The gel filtration profile of ovarian carcinoma tissue homogenate from two patients contained 25 and 1OOkDa peaks of radioligand-bound and immunoreactive folate binding protein, while ascitic fluid from one of the patients exhibited a large 100 kDa immunoreactive peak with no radioligand binding activity. The immunoreactive non-functional 100 kDa FBP could represent unprocessed precursor FBP. Future studies are necessary to evaluate whether determination of immunoreactive FBP in ovarian adenocarcinomatosis is of any diagnostic value. PMID- 9743474 TI - Sendai virus fusion activity as modulated by target membrane components. AB - We have studied the differences between erythrocytes and erythrocyte ghosts as target membranes for the study of Sendai virus fusion activity. Fusion was monitored continuously by fluorescence dequenching of R 18-labeled virus. Experiments were carried out either with or without virus/target membrane prebinding. When Sendai virus was added directly to a erythrocyte/erythrocyte ghost suspension, fusion was always lower than that obtained when experiments were carried out with virus already bound to the erythrocyte/erythrocyte ghost in the cold, since with virus prebinding fusion can be triggered more rapidly. Although virus binding to both erythrocytes and erythrocyte ghosts was similar, fusion activity was much more pronounced when erythrocyte ghosts were used as target membranes. These observations indicate that intact erythrocytes and erythrocyte ghosts are not equivalent as target membranes for the study of Sendai virus fusion activity. Fusion of Sendai virus with both target membranes was inhibited when erythrocytes or erythrocyte ghosts were pretreated with proteinase K, suggesting a role of target membrane proteins in this process. Treatment of both target membranes with neuraminidase, which removes sialic acid residues (the biological receptors for Sendai virus) greatly reduced viral binding. Interestingly, this treatment had no significant effect on the fusion reaction itself. PMID- 9743475 TI - The effects of pH and intraliposomal buffer strength on the rate of liposome content release and intracellular drug delivery. AB - Targeted liposomal drug formulations may enter cells by receptor-mediated endocytosis and then traffick by membrane flow into acidic intracellular compartments. In order to understand the impact of these intracellular pH changes on liposomal drug unloading, the effect of pH on the release from folate-targeted liposomes of three model compounds with distinct pH dependencies was examined. 5(6)-carboxyfluorescein, which titrates from its anionic to uncharged form following internalization by KB cells, displays strong endocytosis-dependent release, since only its uncharged (endosomal) form is membrane permeable. Endocytosis-triggered unloading of drugs of this sort is enhanced by encapsulating the drug in a weak buffer at neutral pH, so that acidification of the intraliposomal compartment following cellular uptake can occur rapidly. Sulforhodamine B, in contrast, retains both anionic and cationic charges at endosomal pH (approximately pH 5), and consequently, escapes the endosomes only very slowly. Doxorubicin, which is commonly loaded into liposomes in its membrane impermeable (cationic) form using an acidic buffer, still displays endocytosis triggered unloading, since sufficient uncharged doxorubicin remains at endosomal pHs to allow rapid re-equilibration of the drug according to the new proton gradient across the membrane. In this case, when the extraliposomal [H+] increases 250-fold from 4 x 10(-8) M (pH 7.4, outside the cell) to 10(-5) M (pH 5, inside the endosome), the ratio of doxorubicin inside to outside the liposome must decrease by a factor of 250. Therefore, the collapse of the transliposomal pH gradient indirectly drives an efflux of the drug molecule from the liposome. Since a change in intraliposomal pH is not required to unload drugs of this type, the intraliposomal compartment can be buffered strongly at acidic pH to prevent premature release of the drug outside the cell. In summary, pH triggered release of liposome-encapsulated drugs can be achieved both with drugs that increase as well as decrease their membrane permeabilities upon acidification, as long as the intraliposomal buffer strength and pH is rationally selected. PMID- 9743476 TI - A novel phosphorylated lipid counteracts activation of the renal plasma membrane (Ca2+ + Mg2+)ATPase by endogenous phosphatidylinositol-4-phosphate. AB - The plasma membrane (Ca2+ + Mg2+)ATPase is activated by acidic phospholipids in reconstituted systems. In this report it is shown that reversible phosphorylation of endogenous phosphatidylinositol regulates the renal plasma membrane (Ca2+ + Mg2+)ATPase, and that a novel phosphorylated lipid that can be isolated from the same membrane strongly counteracts the stimulatory effect of phosphatidylinositol 4-phosphate. PMID- 9743477 TI - Effect of palmitic and lauric acids on the phospholipid bilayer membrane conductivity. AB - Palmitic acid increased the conductivity of BLM from mitochondrial phospholipids when they were dissolved in a mixture of decane and chlorodecane, and was ineffective when phospholipids were dissolved in decane. Lauric acid produced an increase in the membrane conductivity independently of the phospholipid type in the membrane-forming solutions (mitochondrial phospholipids, asolectin, lecithin with cholesterol) and their solvents (decane or decane with chlorodecane). The results show that discrepancies between published data concerning fatty acid effects on the BLM conductivity may be explained by differences in phospholipids, their solvents and fatty acid used. PMID- 9743478 TI - Supplementing state and national health care workforce planning: a regional effort. PMID- 9743479 TI - A multidisciplinary allied health faculty team: formation and first year production of problem-based learning in gerontology/geriatrics. AB - An interdisciplinary team of faculty, administrators and practitioners representing diverse settings for allied health education has formed the Mid Atlantic Allied Health Geriatric Education Center (MAHGEC) to produce problem based learning (PBL) cases related to older adults. These cases will enable allied health students and practitioners to work together in interdisciplinary teams and expand allied health education to include health issues related to gerontology/ geriatrics. The health professionals of MAHGEC have brought different perspectives to the project. These include: (1) educational requirements to be gained from their association with MAHGEC, (2) health care disciplines to enhance the PBL cases produced, (3) personal histories related to older adults, and (4) ideas for utilization of problem-based learning in their particular educational and professional settings. The first year of this project has included the execution of a needs assessment for gerontology/geriatric education in allied health programs, the development of the infrastructure of MAHGEC, building the content base of MAHGEC faculty regarding gerontology/geriatrics and problem-based learning, the establishment of priorities in the production of problem-based learning cases for Year 01, and division into production teams for cases. PMID- 9743480 TI - Improving interdisciplinary team process: a practical approach to team development. PMID- 9743481 TI - Tripartite involvement in health care clinical education. AB - This paper discusses the importance of an appropriate student-clinical educator university lecturer (tripartite) relationship in clinical education and substantiates this importance with the experiences of the dynamics of such relationship in a physiotherapy course. PMID- 9743482 TI - Multiskilled allied health practitioners for Kentucky. PMID- 9743483 TI - Surgical pathophysiology and disease management during the centuries. AB - During the centuries the pathophysiological concept of illness underwent drastic changes, depending on medical discoveries and religious belief. There is evidences of surgical practice in Mesopotamia, Egypt, and Greece 2000 years B.C., but we have to wait the Roman Empire with Celso, Galeno, and most of all Asclepiade and Temisone to appreciate the endeavor to rationally understand diseases as an alteration among different body constituents and not only as divine will. During the Middle Ages, when medicine was disputed by clergy, Ugo and Teodorico Borgognone perceived by intuition the problem of sepsis in patient recovery. This problem was cleared up six centuries later by Semmelweis with his antisepsis. Discoveries of blood circulation, hemostasis, anesthesia, and antisepsis in the 17th and 19th centuries permitted better disease management and have promoted, since the 1950s, pathophysiology as an autonomous discipline. Now pathophysiological mechanisms are used for comprehension of several diseases, such as gallstone formation, peptic ulcers, cranial trauma, cardiovascular abnormalities, and many others, to suggest to us which are the better surgical or clinical trials to realize, while observing caution about their limits and consequences. By a multidisciplinary approach involving surgeons, physiologists, biologists, and physicians, we are able to facilitate patient recovery and to perform modern, scientific, high-quality disease management. PMID- 9743484 TI - Performance of subcutaneously implanted glucose sensors: a review. AB - Despite a considerable amount of research attributed to the development of an implantable glucose sensor, to date there is no clinically applicable concept for continuous glucose monitoring. Investigations to validate the subcutaneous tissue for continuous glucose sensing mostly comprise short-term implantations of glucose sensors. Most implanted glucose sensors showed a significant decay in sensitivity over the implantation period. This bioinstability was not to be expected from the in vitro performance of the sensors. In this article, the influence of possible failure mechanisms on the poor in vivo performance of subcutaneously implanted glucose sensors is reviewed. PMID- 9743485 TI - Long-term evaluation of orthotopically implanted stentless bioprosthetic aortic valves in juvenile sheep. AB - The aim of this study was to develop a technically feasible and reproducible model for chronic evaluation of stentless bioprosthetic aortic valves implanted orthotopically using juvenile domestic sheep. This report summarizes the results of a study conducted to assess orthotopically placed 19-mm stentless aortic bioprosthetic valves. Twenty-seven juvenile sheep underwent aortic valve replacement. Standard cardiopulmonary bypass techniques were followed. The average cardiopulmonary bypass time was 73 min. No chronic anticoagulation was used. There were two deaths (7%) due to surgical complications. In the remaining 25 experiments, 11 animals (41%) died prior to the scheduled sacrifice on postoperative day 150. One early death occurred due to coccidiomycosis infection, one due to technical error, one due to pulmonary embolus, four due to prosthetic annular size disproportion, and four due to thrombi. The remaining 14 animals (52%) underwent left and right heart catheterization, angiography, echocardiography, and sacrifice after postoperative day 150. The average weight of the sheep at elective sacrifice was 60 kg (mean weight gain 12.5 kg). The average cardiac output for the sacrificed animals was 5.1 L/min. The mean velocity of blood across the aortic valve for the sacrificed animals was 317 cm/s and the mean pressure gradient was 26.2 mm Hg. Two features suggest that this model may have broad application. First, we have demonstrated that it is technically feasible to evaluate orthotopically placed stentless bioprosthetic aortic valves in growing sheep. Second, the aortic root size of the juvenile sheep allows for implantation and evaluation of a human size aortic valve (19 mm). We believe that this model is reproducible and can be used to study stentless valve designs. PMID- 9743486 TI - Experimental modifications to a canine infrarenal aortic aneurysm model for the validation of endovascular stent-grafts: an exploratory study. AB - The intraluminal elastase perfusion model has been proven to be potentially effective in producing abdominal aortic aneurysms (AAA) in rodents, yet has produced unpredictable results in larger animals. The purpose of this study was to explore different variations to an existing elastase perfusion model in the dog in the hopes of producing a consistent AAA for endovascular graft validation. The elastase perfusion canine model was modified as follows: (1) inflation of a balloon catheter in the infrarenal aorta (IA) of 3 dogs following elastase perfusion with doses of 2800 U for 40 min; (2) perfusion of the IA of 5 dogs with various elastase doses ranging from 2800 U to 8400 U for 2 h; and (3) perfusion of the IA of 2 dogs with elastase and collagenase for 2 h. The dogs were sacrificed at 4, 7, and 29 weeks. Prior to sacrifice, the treated aortic segments were either examined in vivo by x-ray angiography or by ultrasonography to measure aneurysmal dilation. The aortas were examined macroscopically postmortem to assess the luminal surface characteristics, and under light microscopy and scanning electron microscopy to reveal any pathological injuries induced by the various treatments on the aortic wall. Perfusion of the aorta with 2800 U elastase for 40 min followed by balloon catheter inflation either immediately or 3 weeks after perfusion produced no dilation. Perfusion for 2 h with either elastase alone or in combination with collagenase showed an increased aortic diameter averaging 65.6+/-20.8%, with an irregular dilation of the aortic wall. Histological examination revealed partially digested elastic network of the intima, media, and adventitia, as well as a reduction in the number of smooth muscle cells. An intimal hyperplasic reaction was observed in some of the dogs. Located sparingly within the intima were extravasated erythrocytes associated with recent hemorrhages, intramural thrombi in reorganization, and occasional necrotic lesions. The various modifications brought to the elastase perfusion model failed to produced an aneurysmal dilation with enough expansion to make it a reliable model for endovascular graft validation. PMID- 9743487 TI - Cholecysto-sphincter inhibitory reflex: identification of a reflex and its role in bile flow in a canine model. AB - To study the effect of gallbladder (GB) distension on the sphincter of Oddi (SO), 9 mongrel dogs (mean weight 15.3+/-3.6 kg) were studied. Under anesthesia, the abdomen was opened and the GB and SO were exposed. A balloon-tipped catheter was introduced into the GB and a manometric catheter into the common bile duct so that its fluoroscopically controlled tip lay within the SO. The pressure response of the GB and SO to GB distension by the balloon without and with selective anesthetization of the GB and SO was recorded. The test was repeated in four vagotomized dogs. GB distension effected pressure rise within the GB and pressure drop within the SO. The GB pressure increased progressively as the distending volume increased, while the SO pressure drop was not affected. Selective anesthetization of the GB or the SO produced no SO pressure changes upon GB distension. The SO pressure response to GB distension after vagotomy was similar to that before vagotomy. The SO relaxation on GB contraction, being reproducible and abolished by selective anesthetization of either the SO or the GB, postulates a reflex relationship that we call the cholecysto-sphincter inhibitory reflex. This reflex seems to regulate the bile flow from the GB to the duodenum through the SO. PMID- 9743488 TI - Method for long-term cerebrospinal fluid collection in the conscious dog. AB - In medical research there is often a need for cerebrospinal fluid (CSF) collection from conscious animals. A number of models have been published, but long-term use is often limited by factors such as damage to implant or loss of patency. When part of an implant is exteriorized, group housing is often not possible; even when singly housed, the instrumented animals might have to be fitted with protective devices such as collars, vests, or helmets. Over the last 10 years we have, through the use of this technique, managed to perform repeated CSF collections in 12 beagles. Time of implant duration has ranged from 2.3 to 6.8 years. The method uses two permanent stainless steel guide cannulas screwed into the parietal bone, positioned above but not penetrating the lateral ventricles of the brain. At the time of collection, one of the subcutaneous stainless steel cannulas is used to serve as a guide for a 20-gauge collection needle. The collection is performed under local analgesia and with minimal discomfort to the dog. With its long-term use, we feel that this model is very useful and of high ethical and welfare standards due to the virtual lack of postoperative complications, maintenance requirements, and housing restrictions. PMID- 9743490 TI - Quebrada jaguay: early south american maritime adaptations AB - Excavations at Quebrada Jaguay 280 (QJ-280) (16 degrees30'S) in south coastal Peru demonstrated that Paleoindian-age people of the Terminal Pleistocene (about 11,100 to 10,000 carbon-14 years before the present or about 13,000 to 11,000 calibrated years before the present) in South America relied on marine resources while resident on the coast, which extends the South American record of maritime exploitation by a millennium. This site supports recent evidence that Paleoindian age people had diverse subsistence systems. The presence of obsidian at QJ-280 shows that the inhabitants had contact with the adjacent Andean highlands during the Terminal Pleistocene. PMID- 9743489 TI - Overview and initial results of the very long baseline interferometry space observatory programme AB - High angular resolution images of extragalactic radio sources are being made with the Highly Advanced Laboratory for Communications and Astronomy (HALCA) satellite and ground-based radio telescopes as part of the Very Long Baseline Interferometry (VLBI) Space Observatory Programme (VSOP). VSOP observations at 1.6 and 5 gigahertz of the milli-arc-second-scale structure of radio quasars enable the quasar core size and the corresponding brightness temperature to be determined, and they enable the motions of jet components that are close to the core to be studied. Here, VSOP images of the gamma-ray source 1156+295, the quasar 1548+056, the ultraluminous quasar 0014+813, and the superluminal quasar 0212+735 are presented and discussed. PMID- 9743491 TI - Early maritime economy and El Nino events at quebrada tacahuay, peru AB - The archaeological site of Quebrada Tacahuay, Peru, dates to 12,700 to 12,500 calibrated years before the present (10,770 to 10,530 carbon-14 years before the present). It contains some of the oldest evidence of maritime-based economic activity in the New World. Recovered materials include a hearth, lithic cutting tools and flakes, and abundant processed marine fauna, primarily seabirds and fish. Sediments below and above the occupation layer were probably generated by El Nino events, indicating that El Nino was active during the Pleistocene as well as during the early and middle Holocene. PMID- 9743492 TI - Turbulent transport reduction by zonal flows: massively parallel simulations AB - Three-dimensional gyrokinetic simulations of microturbulence in magnetically confined toroidal plasmas with massively parallel computers showed that, with linear flow damping, an asymptotic residual flow develops in agreement with analytic calculations. Nonlinear global simulations of instabilities driven by temperature gradients in the ion component of the plasma support the view that turbulence-driven fluctuating zonal flows can substantially reduce turbulent transport. Finally, the outstanding differences in the flow dynamics observed in global and local simulations are found to be due to profile variations. PMID- 9743494 TI - Forest fires: An example of self-organized critical behavior AB - Despite the many complexities concerning their initiation and propagation, forest fires exhibit power-law frequency-area statistics over many orders of magnitude. A simple forest fire model, which is an example of self-organized criticality, exhibits similar behavior. One practical implication of this result is that the frequency-area distribution of small and medium fires can be used to quantify the risk of large fires, as is routinely done for earthquakes. PMID- 9743493 TI - Implications of Mars Pathfinder data for the accretion history of the terrestrial planets. AB - Accretion models of the terrestrial planets often assume planetary bulk compositions with nonvolatile element abundance ratios equivalent to those of C1 carbonaceous chondrites. The moment of inertia factor of Mars reported by the Pathfinder team is inconsistent with a bulk planet C1 Fe/Si ratio or Fe content, which suggests that C1 chondrite accretion models are insufficient to explain the formation of Mars and the other terrestrial planets. Future planetary accretion models will have to account for variations in bulk Fe/Si ratios among the terrestrial planets. PMID- 9743495 TI - Molecular assembly and encapsulation directed by hydrogen-bonding preferences and the filling of space. AB - Multiple copies of a molecule, held together in finite aggregates, give rise to properties and functions that are unique to their assembled states. Because these aggregates are held together by weak forces operating over short distances, a premium is placed on complementarity: The molecular surfaces must facilitate specific interactions that direct the assembly to one aggregate rather than another. Hydrogen-bonding preferences can be combined with molecular curvature to favor the assembly of four self-complementary subunits into a pseudo-spherical capsule. Filling the capsule with smaller, complementary molecules provides the final instruction for the assembly process. PMID- 9743496 TI - Inhibition of xenoreactive natural antibody production by retroviral gene therapy. AB - The major barrier to transplantation across discordant species, such as from pig to human, is rejection mediated by xenoreactive natural antibodies (XNA) that bind the carbohydrate epitope Galalpha1-3Galbeta1-4GlcNAc-R (alphaGal) on donor tissues. This epitope is synthesized by the enzyme glucosyltransferase uridine 5' diphosphate galactose:beta-D-galactosyl-1, 4-N-acetyl-D-glucosaminide alpha(1 3)galactosyltransferase (E.C. 2.4.1.151), or simply alphaGT. When a functional alphaGT gene was introduced by retroviral gene transfer into bone marrow cells, alphaGal XNA production in a murine model ceased. Thus, genetic engineering of bone marrow may overcome humoral rejection of discordant xenografts and may be useful for inducing B cell tolerance. PMID- 9743497 TI - Dendritic integration and its role in computing image velocity. AB - The mechanisms underlying visual motion detection can be studied simultaneously in different cell compartments in vivo by using calcium as a reporter of the spatiotemporal activity distribution in single motion-sensitive cells of the fly. As predicted by the Reichardt model, local dendritic calcium signals are found to indicate the direction and velocity of pattern motion but are corrupted by spatial pattern properties. The latter are canceled out by spatial integration, thus leading to a purely directional selective output signal in the axon. These findings attribute a specific computational task to the dendrites of visual interneurons and imply a functional interpretation of dendritic morphology. PMID- 9743498 TI - Aggregation and motor neuron toxicity of an ALS-linked SOD1 mutant independent from wild-type SOD1. AB - Analysis of transgenic mice expressing familial amyotrophic lateral sclerosis (ALS)-linked mutations in the enzyme superoxide dismutase (SOD1) have shown that motor neuron death arises from a mutant-mediated toxic property or properties. In testing the disease mechanism, both elimination and elevation of wild-type SOD1 were found to have no effect on mutant-mediated disease, which demonstrates that the use of SOD mimetics is unlikely to be an effective therapy and raises the question of whether toxicity arises from superoxide-mediated oxidative stress. Aggregates containing SOD1 were common to disease caused by different mutants, implying that coaggregation of an unidentified essential component or components or aberrant catalysis by misfolded mutants underlies a portion of mutant-mediated toxicity. PMID- 9743499 TI - Regulation of meiotic S phase by Ime2 and a Clb5,6-associated kinase in Saccharomyces cerevisiae. AB - Cyclin-dependent kinase (Cdk) mutations that prevent entry into the mitotic cell cycle of budding yeast fail to block meiotic DNA replication, suggesting there may be fundamental differences between these pathways. However, S phase in meiosis was found to depend on the same B-type cyclins (Clb5 and Clb6) as it does in mitosis. Meiosis differs instead in the mechanism that controls removal of the Cdk inhibitor Sic1. Destruction of Sic1 and activation of a Clb5-dependent kinase in meiotic cells required the action of the meiosis-specific protein kinase Ime2, thereby coupling early meiotic gene expression to control of DNA replication for meiosis. PMID- 9743500 TI - Fertilization defects in sperm from mice lacking fertilin beta. AB - Fertilin, a member of the ADAM family, is found on the plasma membrane of mammalian sperm. Sperm from mice lacking fertilin beta were shown to be deficient in sperm-egg membrane adhesion, sperm-egg fusion, migration from the uterus into the oviduct, and binding to the egg zona pellucida. Egg activation was unaffected. The results are consistent with a direct role of fertilin in sperm egg plasma membrane interaction. Fertilin could also have a direct role in sperm zona binding or oviduct migration; alternatively, the effects on these functions could result from the absence of fertilin activity during spermatogenesis. PMID- 9743501 TI - Activation of apoptosis signal-regulating kinase 1 (ASK1) by the adapter protein Daxx. AB - The Fas death receptor can activate the Jun NH2-terminal kinase (JNK) pathway through the receptor-associated protein Daxx. Daxx was found to activate the JNK kinase kinase ASK1, and overexpression of a kinase-deficient ASK1 mutant inhibited Fas- and Daxx-induced apoptosis and JNK activation. Fas activation induced Daxx to interact with ASK1, which consequently relieved an inhibitory intramolecular interaction between the amino- and carboxyl-termini of ASK1, activating its kinase activity. The Daxx-ASK1 connection completes a signaling pathway from a cell surface death receptor to kinase cascades that modulate nuclear transcription factors. PMID- 9743503 TI - More cellular signals for atherogenesis? PMID- 9743502 TI - Promotion of dendritic growth by CPG15, an activity-induced signaling molecule. AB - Activity-independent and activity-dependent mechanisms work in concert to regulate neuronal growth, ensuring the formation of accurate synaptic connections. CPG15, a protein regulated by synaptic activity, functions as a cell surface growth-promoting molecule in vivo. In Xenopus laevis, CPG15 enhanced dendritic arbor growth in projection neurons, with no effect on interneurons. CPG15 controlled growth of neighboring neurons through an intercellular signaling mechanism that requires its glycosylphosphatidylinositol link. CPG15 may represent a new class of activity-regulated, membrane-bound, growth-promoting proteins that permit exquisite spatial and temporal control of neuronal structure. PMID- 9743504 TI - Cyclosporin A inhibits apoptosis of human endothelial cells by preventing release of cytochrome C from mitochondria. AB - BACKGROUND: Several experimental and clinical studies suggest that cyclosporin A (CSA) treatment reduces transplant atherosclerosis. Because oxidized LDL (oxLDL) is believed to play a key role in the development of atherogenesis, causing injury to the endothelium, and has been shown to induce apoptosis of endothelial cells, we investigated whether CSA inhibits oxLDL-induced apoptosis. METHODS AND RESULTS: Apoptosis was induced in human umbilical venous endothelial cells (HUVECs) by incubation of 10 microg/mL oxLDL for 18 hours. Coincubation with CSA dose dependently decreased oxLDL-induced apoptosis, with a maximal effect at 10 micromol/L. In addition, tumor necrosis factor-alpha- and angiotensin II-induced apoptosis was significantly prevented by CSA treatment, suggesting a general apoptosis-suppressive effect of CSA. CSA has been shown to inhibit disruption of the mitochondrial membrane function, which plays a key role in apoptosis induction. Indeed, oxLDL treatment triggered the release of cytochrome C from the mitochondria into the cytosol, indicating disturbance of the mitochondrial membrane. CSA (10 micromol/L) completely inhibited the oxLDL-induced release of cytochrome C. Moreover, tumor necrosis factor-alpha- and angiotensin II-induced cytochrome C release was prevented by CSA treatment. CONCLUSIONS: OxLDL induces dysfunction of the mitochondrial membrane, leading to cytochrome C release into the cytosol, and thereby stimulates apoptosis of human endothelial cells. Apoptosis suppression by CSA correlates with the prevention of mitochondrial dysfunction and thus indicates the importance of mitochondrial destabilization in oxLDL-induced apoptosis signaling. The inhibition of apoptosis by CSA might preserve the function of the endothelium and may at least in part contribute to the antiatherogenic effects of CSA in transplant atherosclerosis. PMID- 9743505 TI - Estradiol therapy combined with progesterone and endothelium-dependent vasodilation in postmenopausal women. AB - BACKGROUND: Epidemiological studies indicate that estrogen replacement therapy decreases the risk of cardiovascular events in postmenopausal women. Estrogen may confer cardiovascular protection by improving endothelial function because it increases endothelium-dependent vasodilation. It is not known whether progesterone attenuates the beneficial effects of estrogen on endothelial function. METHODS AND RESULTS: Seventeen postmenopausal women with mild hypercholesterolemia were enrolled in a placebo-controlled, crossover trial to evaluate the effect of transdermal estradiol, with and without vaginal micronized progesterone, on endothelium-dependent vasodilation in a peripheral conduit artery. Brachial artery diameter was measured with high-resolution B-mode ultrasonography. To assess endothelium-dependent vasodilation, brachial artery diameter was determined at baseline and after a flow stimulus induced by reactive hyperemia. To assess endothelium-independent vasodilation, brachial artery diameter was measured after administration of sublingual nitroglycerin. During estradiol therapy, reactive hyperemia caused an 11.1+/-1.0% change in brachial artery diameter compared with 4. 7+/-0.6% during placebo therapy (P<0.001). Progesterone did not significantly attenuate this improvement. During combined estrogen and progesterone therapy, flow-mediated vasodilation of the brachial artery was 9.6+/-0.8% (P=NS versus estradiol alone). Endothelium-independent vasodilation was not altered by estradiol therapy, either with or without progesterone, compared with placebo. There was a modest decrease in total and LDL cholesterol during treatment both with estradiol alone and when estradiol was combined with progesterone (all P<0.001 versus placebo). In a multivariate analysis that included serum estradiol, progesterone, total and LDL cholesterol concentrations, blood pressure, and heart rate, only the estradiol level was a significant predictor of endothelium-dependent vasodilation. CONCLUSIONS: The addition of micronized progesterone does not attenuate the favorable effect of estradiol on endothelium-dependent vasodilation. The vasoprotective effect of hormone replacement therapy may extend beyond its beneficial actions on lipids. PMID- 9743506 TI - Activated platelets induce monocyte chemotactic protein-1 secretion and surface expression of intercellular adhesion molecule-1 on endothelial cells. AB - BACKGROUND: Platelet/endothelium interaction plays an important role in the pathophysiology of inflammation and atherosclerosis. The role of platelets for monocyte chemotactic protein-1 (MCP-1) secretion and surface expression of intercellular adhesion molecule-1 (ICAM-1) on endothelial cells has been assessed. METHODS AND RESULTS: Monolayers of human umbilical vein endothelial cells were incubated with nonstimulated or ADP-activated platelets for 6 hours, and secretion of MCP-1 and surface expression of ICAM-1 were determined by ELISA and flow cytometry, respectively. In the presence of ADP-activated platelets, both MCP-1 secretion and ICAM-1 surface expression were significantly increased compared with nonstimulated platelets (P<0.02). Activation of the transcription factor nuclear factor-kappaB (NF-kappaB) determined by electrophoretic mobility shift assay and kappaB-dependent transcriptional activity was enhanced in the presence of activated platelets. In addition, ADP-activated platelets induced MCP 1 and ICAM-1 promoter-dependent transcription. Liposomal transfection of a double stranded kappaB phosphorothioate oligonucleotide, but not of the mutated form, inhibited MCP-1 secretion and surface expression of ICAM-1 on activated endothelium (P<0.05). CONCLUSIONS: The present study indicates that activated platelets modulate chemotactic (MCP-1) and adhesive (ICAM-1) properties of endothelial cells via an NF-kappaB-dependent mechanism. Platelet-induced activation of the NF-kappaB system might contribute to early inflammatory events in atherogenesis. PMID- 9743507 TI - Plasma lipoprotein(a) is not a predictor for restenosis after elective high pressure coronary stenting. AB - BACKGROUND: Lipoprotein(a) is a risk factor for coronary artery disease. Although it has been implicated in restenosis after balloon angioplasty, its role in restenosis within coronary stents is unknown. The aim of the study was to assess the role of plasma lipoprotein(a) as a predictor for restenosis after elective coronary stenting. METHODS AND RESULTS: Elective, high-pressure stenting of de novo lesions in native coronary arteries with Palmaz-Schatz stents was performed in 325 consecutive patients. Clinical, angiographic, and biochemical data were analyzed prospectively. Angiographic follow-up was performed at 6 months. Lipoprotein(a) levels were compared in patients with and without restenosis. Angiographic follow-up was obtained in 312 patients (96%); recurrence was observed in 67 patients (21.5%). No clinical or biochemical variable was associated with restenosis. Lipoprotein(a) level was 37.81+/-49. 01 mg/dL (median, 22 mg/dL; range, 3 to 262 mg/dL) in restenotic patients and 36.95+/ 40.65 mg/dL (median, 22 mg/dL; range, 0 to 244 mg/dL) in nonrestenotic patients (P=NS). The correlations between percent diameter stenosis, minimum luminal diameter, and late loss at follow-up angiography and basal lipoprotein(a) plasma level after logarithmic transformation were 0.006, 0.002, and 0.0017, respectively. Multiple stents were associated with a higher incidence of restenosis (P=0.006), but biochemical data in these patients were similar to those treated with single stents. CONCLUSIONS: The basal plasma level of lipoprotein(a) measured before the procedure is not a predictor for restenosis after elective high-pressure coronary stenting. PMID- 9743509 TI - Clinical effects of beta-adrenergic blockade in chronic heart failure: a meta analysis of double-blind, placebo-controlled, randomized trials. AB - BACKGROUND: beta-Blockers have improved symptoms and reduced the risk of cardiovascular events in studies of patients with heart failure, but it is unclear which end points are most sensitive to the therapeutic effects of these drugs. METHODS AND RESULTS: We combined the results of all 18 published double blind, placebo-controlled, parallel-group trials of beta-blockers in heart failure. From this combined database of 3023 patients, we evaluated the strength of evidence supporting an effect of treatment on left ventricular ejection fraction, NYHA functional class, hospitalizations for heart failure, and death. beta-Blockers exerted their most persuasive effects on ejection fraction and on the combined risk of death and hospitalization for heart failure. beta-Blockade increased the ejection fraction by 29% (P<10(-9)) and reduced the combined risk of death or hospitalization for heart failure by 37% (P<0.001). Both effects remained significant even if >90% of the trials were eliminated from the analysis or if a large number of trials with a neutral result were added to the analysis. In contrast, the effect of beta-blockade on NYHA functional class was of borderline significance (P=0.04) and disappeared with the addition or removal of only 1 moderate-size study. Although beta -blockade reduced all-cause mortality by 32% (P=0.003), this effect was only moderately robust and varied according to the type of ss-blocker tested, ie, the reduction of mortality risk was greater for nonselective beta-blockers than for beta1-selective agents (49% versus 18%, P=0.049). However, selective and nonselective beta-blockers did not differ in their effects on other measures of clinical efficacy. CONCLUSIONS: These analyses indicate that there is persuasive evidence supporting a favorable effect of beta blockade on ejection fraction and the combined risk of death and hospitalization for heart failure. In contrast, the effect of these drugs on other end points requires additional study. PMID- 9743508 TI - Comparison of exercise performance in patients with chronic severe heart failure versus left ventricular assist devices. AB - BACKGROUND: Left ventricular assist devices (LVADs) are frequently used as a bridge to cardiac transplantation and may be useful as long-term therapy. The purpose of this study was to compare the exercise performance of LVAD patients with that of ambulatory heart failure patients. METHODS AND RESULTS: Exercise testing with hemodynamic and respiratory gas measurements was performed in 65 congestive heart failure (CHF) patients (age 53+/-10 years) and 20 LVAD patients (age 49+/-8 years). Peak Vo2 was significantly higher in the LVAD than the CHF patients (CHF, 12+/-3; LVAD, 15. 9+/-3.8 mL . kg-1 . min-1; P<0.001), as was the Vo2 at the anaerobic threshold (CHF, 8.1+/-2.1; LVAD, 12.2+/-2.9 mL . kg-1 . min 1; P<0.001). At rest, mean arterial blood pressure (CHF, 87+/-11; LVAD, 94+/-9 mm Hg) and cardiac output (CHF, 4+/-1; LVAD, 4. 9+/-0.9 L/min) were increased, whereas mean pulmonary artery pressure (CHF, 28+/-11; LVAD, 18+/-4 mm Hg) and pulmonary artery wedge pressure (CHF, 16+/-10; LVAD 5+/-3 mm Hg) were reduced (all P<0.01). At peak exercise, heart rate (CHF,125+/-24; LVAD, 148+/-24 bpm), blood pressure (CHF, 87+/-14; LVAD,96+/-12 mm Hg), and cardiac output (CHF, 7.6+/ 2.2; LVAD, 11.2+/-2.6 L/min) were higher (all P<0. 01), whereas mean pulmonary artery pressure (CHF, 48+/-12; LVAD, 30+/-5 mm Hg) and mean pulmonary capillary wedge pressure (CHF, 31+/-11; LVAD, 14+/-6 mm Hg) were lower in the LVAD group (both P<0. 001). In the LVAD patients, Fick cardiac output was higher than LVAD flow sensor value measurements (Fick, 11.6+/-2.5; LVAD, 8.1+/-1.2 L/min; P<0.001). CONCLUSIONS: Hemodynamic measurements at rest and during exercise are significantly improved in patients with devices compared with those in ambulatory heart failure patients awaiting cardiac transplantation. Similarly, the exercise capacity of device patients is better than that of transplant candidates and in the majority of patients is similar to that of patients with mild CHF. PMID- 9743510 TI - Percutaneous transluminal coronary angioplasty reverses vasoconstriction of stenotic coronary arteries in hypertensive patients. AB - BACKGROUND: Endothelial dysfunction of coronary arteries with impaired vasodilation has been reported in patients with arterial hypertension. However, the effect of dynamic exercise on coronary vasomotion of a stenotic vessel segment before and after PTCA has not yet been evaluated in these patients. METHODS AND RESULTS: Coronary vasomotion of a normal and a stenotic vessel segment was studied in 39 patients with coronary artery disease during supine bicycle exercise before and 9+/-3 months after PTCA. Luminal area changes were determined by biplane quantitative coronary arteriography. There were 21 normotensive and 18 hypertensive patients who did not differ with regard to clinical characteristics. Percent area stenosis decreased after PTCA from 90% to 39% (P<0.001) in normotensive and from 86% to 33% (P<0.001) in hypertensive patients. Exercise-induced vasomotion of the normal vessel segment was significantly different between normotensives and hypertensives before (+19% versus +1%, P<0.01) and after (+16% versus +3%, P<0.01) PTCA. In contrast, stenotic vessel segments showed vasoconstriction in both normotensive and hypertensive patients (Deltaexercise, -11% versus - 20%, P=NS), which was reversed after PTCA (+3% versus +2%, P=NS). CONCLUSIONS: Normal coronary arteries show reduced vasodilation during exercise in hypertensive patients that may be explained by the presence of endothelial dysfunction. Stenotic vessels demonstrate paradoxical vasoconstriction during exercise in both normotensive and hypertensive patients. PTCA reverses vasoconstriction by elimination of the flow limiting stenosis and prevention of coronary stenosis narrowing during exercise in normotensive and hypertensive patients. PMID- 9743512 TI - Application of color Doppler flow mapping to calculate orifice area of St Jude mitral valve. AB - BACKGROUND: The effective orifice area (EOA) of a prosthetic valve is superior to transvalvular gradients as a measure of valve function, but measurement of mitral prosthesis EOA has not been reliable. METHODS AND RESULTS: In vitro flow across St Jude valves was calculated by hemispheric proximal isovelocity surface area (PISA) and segment-of-spheroid (SOS) methods. For steady and pulsatile conditions, PISA and SOS flows correlated with true flow, but SOS and not PISA underestimated flow. These principles were then used intraoperatively to calculate cardiac output and EOA of newly implanted St Jude mitral valves in 36 patients. Cardiac output by PISA agreed closely with thermodilution (r=0.91, Delta=-0.05+/-0.55 L/min), but SOS underestimated it (r=0.82, Delta=-1.33+/-0.73 L/min). Doppler EOAs correlated with Gorlin equation estimates (r=0.75 for PISA and r=0.68 for SOS, P<0.001) but were smaller than corresponding in vitro EOA estimates. CONCLUSIONS: Proximal flow convergence methods can calculate forward flow and estimate EOA of St Jude mitral valves, which may improve noninvasive assessment of prosthetic mitral valve obstruction. PMID- 9743511 TI - Intake of potassium, magnesium, calcium, and fiber and risk of stroke among US men. AB - BACKGROUND: Animal experiments and epidemiological studies have suggested that high potassium intake may reduce the risk of stroke, but the evidence is inconclusive, and the role of other nutrients in potassium-rich foods remains unknown. METHODS AND RESULTS: We examined the association of potassium and related nutrients with risk of stroke among 43 738 US men, 40 to 75 years old, without diagnosed cardiovascular diseases or diabetes, who completed a semiquantitative food frequency questionnaire in 1986. During 8 years of follow up, 328 strokes (210 ischemic, 70 hemorrhagic, 48 unspecified) were documented. The multivariate relative risk of stroke of any type for men in the top fifth of potassium intake (median intake, 4.3 g/d) versus those in the bottom (median, 2.4 g/d) was 0.62 (95% CI, 0.43, 0.88; P for trend=0.007). Results for ischemic stroke alone were similar. Intakes of cereal fiber and magnesium, but not of calcium, were also inversely associated with risk of total stroke. These inverse associations were all stronger in hypertensive than normotensive men and were not materially altered by adjustment for blood pressure levels. Use of potassium supplements was also inversely related to risk of stroke, particularly among men taking diuretics (relative risk, 0.36; 95% CI, 0.18, 0.72). CONCLUSIONS: Although these data do not prove a causal relationship, they are consistent with the hypothesis that diets rich in potassium, magnesium, and cereal fiber reduce the risk of stroke, particularly among hypertensive men. Potassium supplements may also be beneficial, but because of potential risks, use should be carefully monitored and restricted to men taking potassium-losing diuretics. PMID- 9743513 TI - Transfection of inducible nitric oxide synthase gene causes apoptosis in vascular smooth muscle cells. AB - BACKGROUND: Excess production of nitric oxide (NO) by inducible NO synthase (iNOS) has been implicated in a variety of physiological processes including vascular remodeling. To elucidate whether endogenous NO generated by iNOS is involved in the programmed cell death (apoptosis) of the vasculature, iNOS cDNA- expressing construct was transfected into rat and human vascular smooth muscle cells (VSMCs) by lipofection. METHODS AND RESULTS: VSMCs transiently transfected with iNOS cDNA functionally expressed 130 kd iNOS protein with full catalytic activity to generate massive NO in proportion to the doses of cDNA used; its enzymatic activity as well as NO production was completely blocked by an NOS inhibitor, NG-monomethyl-L-arginine (LNMMA). Overexpression of iNOS led to a marked inhibition of DNA synthesis as well as induction of apoptosis in VSMCs. Evidence for apoptotic cell death was provided by internucleosomal DNA fragmentation by agarose gel electrophoresis, positive staining for TdT-mediated dUTP biotin nick end-labeling, and appearance of hypodiploid cells by flow cytometry analysis. Apoptosis after transfection with iNOS cDNA was abrogated by LNMMA. Transfection of iNOS cDNA caused accumulation of the tumor suppressor gene p53 but not of bcl-2, which was also blocked by LNMMA. CONCLUSIONS: These results demonstrate that massive generation of endogenous NO derived from iNOS overexpression leads to a marked apoptosis in VSMCs, thus suggesting an important role of NO as a proapoptotic factor for VSMCs in the process of vascular remodeling. PMID- 9743514 TI - all-Trans-retinoic acid reduces neointimal formation and promotes favorable geometric remodeling of the rat carotid artery after balloon withdrawal injury. AB - BACKGROUND: The multifactorial and unpredictable nature of human restenosis will probably necessitate interventional strategies that target multiple processes involved in acute vascular narrowing. Retinoids (eg, all-trans-retinoic acid, atRA) represent a growing class of pleiotropic biological response modifiers with demonstrable efficacy in managing several pathological conditions. In this report, we have initiated studies to examine the hypothesis that atRA limits neointimal formation after experimental vascular injury. METHODS AND RESULTS: Rats were predosed with atRA (30 mg . kg-1 . d-1 PO) or corn oil 4 days before balloon withdrawal injury (BWI) of the left common carotid artery and continued on this drug regimen for an additional 14 days. High-performance liquid chromatographic analysis documented therapeutic levels of atRA in serum and vascular tissue. atRA depressed peak DNA synthesis in the tunica media of BWI vessels (P<0.05). Histomorphometry revealed atRA-mediated reductions in neointimal area, neointimal cell number, and intimal/medial area ratio as well as significant increases in vessel wall perimeter (P<0. 05). Such changes in vascular architecture contributed to a 35% to 37% increase in the luminal area of BWI vessels exposed to atRA (P<0. 005 compared with controls). CONCLUSIONS: atRA reduces neointimal mass and elicits favorable geometric remodeling of the injured rat carotid artery. PMID- 9743515 TI - Role of adenosine receptors in the paradoxic bradycardia response of rats to inferior vena cava occlusion during an infusion of isoproterenol. AB - BACKGROUND: In susceptible humans, vasodepressor reactions are induced by restriction of venous return (upright tilting) and administration of isoproterenol. Because paradoxic bradycardia is a major manifestation of vasodepressor reactions, and allowing for extrapolation between paradoxic bradycardia in rats and vasodepressor reactions, we examined whether adenosine receptors mediate the paradoxic bradycardia reaction. METHODS AND RESULTS: Paradoxic bradycardia was induced in rats by inferior vena cava occlusion during an isoproterenol infusion. We studied whether dipyridamole, an adenosine transport inhibitor, and aminophylline (nonselective) or DPCPX (selective) A1 antagonists augmented or inhibited paradoxic bradycardia, respectively, during inferior vena cava occlusion. The maximum changes in R-R during 60 seconds of inferior vena cava occlusion were that (1) in control, the rate accelerated (DeltaR-R, -9.7+/-0.8 ms, P<0.001); (2) during isoproterenol (0.8 microg . min 1), paradoxic bradycardia occurred (DeltaR-R, +92.0+/-32.0 ms, P<0.001); (3) during isoproterenol but after dipyridamole, paradoxic bradycardia occurred at a much lower dose of isoproterenol (0.2 microg . min-1), and the magnitude was increased at all doses (at 0.8 microg . min-1 isoproterenol, DeltaR-R, +195.6+/ 27.6 ms, P<0.001 versus isoproterenol alone, DeltaR-R, +92.0+/-32 ms); (4) during isoproterenol and dipyridamole, atropine did not block paradoxic bradycardia, but cervical vagotomy inhibited paradoxic bradycardia (DeltaR-R, +5.6+/-1.8 ms, P<0.001 compared with isoproterenol and dipyridamole alone); and (5) during isoproterenol alone, aminophylline or DPCPX blocked paradoxic bradycardia (DeltaR R, -5.4+/-1.0 ms, and DeltaR-R, -2.6+/-0.5 ms, respectively, each P<0.001 compared with isoproterenol alone). CONCLUSIONS: The adenosine A1 receptor mediates the paradoxic bradycardia reflex during inferior vena cava occlusion in the face of isoproterenol via vagal afferents. PMID- 9743516 TI - Spatiotemporal periodicity during atrial fibrillation in the isolated sheep heart. AB - BACKGROUND: The activation patterns that underlie the irregular electrical activity during atrial fibrillation (AF) have traditionally been described as disorganized or random. Recent studies, based predominantly on statistical methods, have provided evidence that AF is spatially organized. The objective of this study was to demonstrate the presence of spatial and temporal periodicity during AF. METHODS AND RESULTS: We used a combination of high-resolution video imaging, ECG recordings, and spectral analysis to identify sequential wave fronts with temporal periodicity and similar spatial patterns of propagation during 20 episodes of AF in 6 Langendorff-perfused sheep hearts. Spectral analysis of AF demonstrated multiple narrow-band peaks with a single dominant peak in all cases (mean, 9.4+/-2.6 Hz; cycle length, 112+/-26 ms). Evidence of spatiotemporal periodicity was found in 12 of 20 optical recordings of the right atrium (RA) and in all (n=19) recordings of the left atrium (LA). The cycle length of spatiotemporal periodic waves correlated with the dominant frequency of their respective optical pseudo-ECGs (LA: R2=0.99, slope=0.94 [95% CI, 0.88 to 0.99]; RA: R2=0.97, slope=0.92 [95% CI, 0.80 to 1.03]). The dominant frequency of the LA pseudo-ECG alone correlated with the global bipolar atrial EG (R2=0.76, slope=0.75 [95% CI, 0.52 to 0.99]). In specific examples, sources of periodic activity were seen as rotors in the epicardial sheet or as periodic breakthroughs that most likely represented transmural pectinate muscle reentry. However, in the majority of cases, periodic waves were seen to enter the mapping area from the edge of the field of view. CONCLUSIONS: Reentry in anatomically or functionally determined circuits forms the basis of spatiotemporal periodic activity during AF. The cycle length of sources in the LA determines the dominant peak in the frequency spectra in this experimental model of AF. PMID- 9743517 TI - Platelet glycoprotein IIb/IIIa receptor blockade in acute myocardial infarction associated with thrombotic occlusion of the left main coronary artery. PMID- 9743518 TI - Papillary fibroelastoma of the mitral valve with systemic embolization. PMID- 9743519 TI - Translation of a retained intron in tyrosinase-related protein (TRP) 2 mRNA generates a new cytotoxic T lymphocyte (CTL)-defined and shared human melanoma antigen not expressed in normal cells of the melanocytic lineage. AB - We report here the identification of a new shared human melanoma antigen recognized by a human leukocyte antigen (HLA)-A*68011-restricted cytotoxic T lymphocyte clone (CTL 128). The cDNA encoding this antigen is composed of a partially spliced form of the melanocyte differentiation antigen tyrosinase related protein (TRP)-2, containing exons 1-4 with retention of intron 2 and part of intron 4 (TRP-2-INT2). The sequence coding for the antigenic epitope is located at the 5' end of intron 2 and is available for translation in the same open reading frame of the fully spliced TRP-2 mRNA. This peptide is also recognized by CTL 128 when presented by the HLA-A*3301, a member of the HLA-A3 like supertype that includes the HLA-A*68011. Quantitative reverse transcription PCR analysis carried out on total and/or cytoplasmic mRNA demonstrated that, in contrast to the fully spliced TRP-2 mRNA expressed in melanomas, normal skin melanocytes, and retina, the TRP-2-INT2 mRNA could be detected at significant levels in melanomas but not in normal cells of the melanocytic lineage. Instead, in these normal samples, both the spliced and the unspliced transcript of gp100 were expressed at high levels. Absence of endogenous TRP-2-INT2 expression in melanocytes was also confirmed by lack of recognition of HLA-A*68011-transduced, TRP-2(+) melanocyte lines by CTL 128. These results indicate that a partially spliced form of a differentiation antigen mRNA, present in the cytoplasmic compartment of neoplastic but not normal cells of the melanocytic lineage, can be the source of a melanoma-restricted T cell epitope. PMID- 9743520 TI - The sequence alteration associated with a mutational hotspot in p53 protects cells from lysis by cytotoxic T lymphocytes specific for a flanking peptide epitope. AB - A high proportion of tumors arise due to mutation of the p53 tumor suppressor protein. A p53 hotspot mutation at amino acid position 273 from R to H, flanking a peptide epitope that spans residues 264-272, renders cells resistant to killing by human histocompatibility leukocyte antigen (HLA)-A*0201-restricted cytotoxic T lymphocytes (CTLs) specific for this epitope. Acquisition of the R to H mutation at residue 273 of the human p53 protein promotes tumor growth in vivo by selective escape from recognition by p53.264-272 peptide-specific CTLs. Synthetic 27-mer p53 polypeptides covering the antigenic nonamer region 264-272 of p53 were used as proteasome substrates to investigate whether the R to H mutation at the P1' position of the COOH terminus of the epitope affects proteasome-mediated processing of the protein. Analysis of the generated products by tandem mass spectrometry and the kinetics of polypeptide processing in conjunction with CTL assays demonstrate that the R to H mutation alters proteasomal processing of the p53 protein by inhibiting proteolytic cleavage between residues 272 and 273. This prevents the release of the natural CTL epitope that spans flanking residues 264 272 as well as a putative precursor peptide. These results demonstrate that mutation of p53 not only leads to malignant transformation but may also, in some instances, affect immune surveillance and should be considered in the design of cancer vaccines. PMID- 9743521 TI - Urokinase receptor (CD87) regulates leukocyte recruitment via beta 2 integrins in vivo. AB - The urokinase receptor (CD87; uPAR) is found in close association with beta 2 integrins on leukocytes. We studied the functional consequence of this association for leukocyte adhesion and migration. In vivo, the beta 2 integrin dependent recruitment of leukocytes to the inflamed peritoneum of uPAR-deficient mice was significantly reduced as compared with wild-type animals. In vitro, beta 2 integrin-mediated adhesion of leukocytes to endothelium was lost upon removal of uPAR from the leukocyte surface by phosphatidyl-inositol-specific phospholipase C. Leukocyte adhesion was reconstituted when soluble intact uPAR, but not a truncated form lacking the uPA-binding domain, was allowed to reassociate with the cell surface. uPAR ligation with a monoclonal antibody induced adhesion of monocytic cells and neutrophils to vascular endothelium by six- to eightfold, whereas ligation with inactivated uPA significantly reduced cell-to-cell adhesion irrespective of the beta 2 integrin-stimulating pathway. These data indicate that beta 2 integrin-mediated leukocyte-endothelial cell interactions and recruitment to inflamed areas require the presence of uPAR and define a new phenotype for uPAR-deficient mice. Moreover, uPAR ligation differentially modulates leukocyte adhesion to endothelium and provides novel targets for therapeutic strategies in inflammation-related vascular pathologies. PMID- 9743522 TI - Inhibition of angiogenesis by interleukin 4. AB - Interleukin (IL)-4, a crucial modulator of the immune system and an active antitumor agent, is also a potent inhibitor of angiogenesis. When incorporated at concentrations of 10 ng/ml or more into pellets implanted into the rat cornea or when delivered systemically to the mouse by intraperitoneal injection, IL-4 blocked the induction of corneal neovascularization by basic fibroblast growth factor. IL-4 as well as IL-13 inhibited the migration of cultured bovine or human microvascular cells, showing unusual dose-response curves that were sharply stimulatory at a concentration of 0.01 ng/ml but inhibitory over a wide range of higher concentrations. Recombinant cytokine from mouse and from human worked equally well in vitro on bovine and human endothelial cells and in vivo in the rat, showing no species specificity. IL-4 was secreted at inhibitory levels by activated murine T helper (TH0) cells and by a line of carcinoma cells whose tumorigenicity is known to be inhibited by IL-4. Its ability to cause media conditioned by these cells to be antiangiogenic suggested that the antiangiogenic activity of IL-4 may play a role in normal physiology and contribute significantly to its demonstrated antitumor activity. PMID- 9743523 TI - Hierarchical and redundant lymphocyte subset control precludes cytomegalovirus replication during latent infection. AB - Reactivation from latent cytomegalovirus (CMV) infection is often associated with conditions of immunosuppression and can result in fatal disease. Whether the maintenance of systemic CMV latency is mainly governed by factors of the infected cell or by immune control functions is unknown. Likewise, the putative immune control mechanisms which could prevent the induction and spread of recurrent CMV infection are not clearly identified. We took advantage of latently infected B cell-deficient mice and a sensitive method for virus detection to study CMV reactivation after ablation of lymphocyte subsets. A crucial role of both T lymphocytes and natural killer (NK) cells was demonstrated. Within 5 d after depletion of lymphocytes, productive infection occurred in 50% of mice, and 14 d later 100% of mice exhibited recurrent infection. A hierarchy of immune control functions of CD8(+), NK, and CD4(+) cells was established. Reactivation was rare if only one of the lymphocyte subsets was depleted, but was evident after removal of a further subset, indicating a functional redundancy of control mechanisms. The salivary glands were identified as the site of most rapid virus shedding, followed by the detection of recurrent virus in the lungs, and eventually in the spleen. Our findings document a previously unknown propensity of latent CMV genomes to enter productive infection immediately and with a high frequency after immune cell depletion. The data indicate that only the sustained cellular immune control prevents CMV replication and restricts the viral genome to a systemic state of latency. PMID- 9743524 TI - Functional redundancy of the nuclear factor kappa B inhibitors I kappa B alpha and I kappa B beta. AB - The transcription factor NF-kappaB is sequestered in the cytoplasm by the inhibitor proteins of the IkappaB family. Each member of the IkappaB exhibits structural and biochemical similarities as well as differences. In an effort to address the functional redundancy of two closely related IkappaB molecules, IkappaBalpha and IkappaBbeta, we generated knock-in mice by replacing the IkappaBalpha gene with the IkappaBbeta gene. The knock-in mice do not express IkappaBalpha, but express a T7-tagged IkappaBbeta under the promoter and regulatory sequence of ikba. Unlike the IkappaBalpha-deficient mice, which display severe postnatal developmental defects and die by postnatal day 8, homozygous knock-in mice survive to adulthood, are fertile, and exhibit no apparent abnormalities. Furthermore, thymocytes and embryonic fibroblasts from the knock-in animals exhibit an inducible NF-kappaB response similar to that of wild-type animals. These results indicate that IkappaBalpha and IkappaBbeta share significant similarities in their biochemical activity, and that they acquired their different functions from divergent expression patterns during evolution. PMID- 9743525 TI - Molecular and biological characterization of the murine leukotriene B4 receptor expressed on eosinophils. AB - The movement of leukocytes into tissues is regulated by the local production of chemical mediators collectively referred to as chemoattractants. Although chemoattractants constitute a diverse array of molecules, including proteins, peptides, and lipids, they all appear to signal leukocytes through a related family of seven transmembrane-spanning G protein-coupled receptors. The eosinophil is a potent proinflammatory cell that is attracted into tissues during allergic inflammation, parasitic infection, and certain malignancies. Since the molecular mechanisms controlling eosinophil recruitment are incompletely understood, we performed a degenerate polymerase chain reaction on cDNA isolated from murine eosinophils to identify novel chemoattractant receptors. We report the isolation of a cDNA that encodes a 351-amino acid glycoprotein that is 78% identical to a human gene that has been reported to be a purinoceptor (P2Y7) and a leukotriene B4 (LTB4) receptor (BLTR). Chinese hamster ovary (CHO) cells transfected with this cDNA specifically bound [3H]LTB4 with a dissociation constant of 0.6 +/- 0.1 nM. Furthermore, LTB4 induced a dose-dependent intracellular calcium flux in transfected CHO cells. In contrast, [35S]dATP did not specifically bind to these transfectants. This mRNA was expressed at high levels in interleukin 5-exposed eosinophils, elicited peritoneal macrophages and neutrophils, and to a lesser extent interferon gamma stimulated macrophages. Low levels of expression were detected in the lung, lymph node, and spleen of unchallenged mice. Western blot analysis detected the mBLTR protein in murine eosinophils and alveolar macrophages as well as human eosinophils. In addition, elevated levels of mBLTR mRNA were found in the lungs of mice in a murine model of allergic pulmonary inflammation in a time course consistent with the influx of eosinophils. Our findings indicate that this murine receptor is an LTB4 receptor that is highly expressed on activated leukocytes, including eosinophils, and may play an important role in mediating eosinophil recruitment into inflammatory foci. PMID- 9743526 TI - Predominant role for directly transfected dendritic cells in antigen presentation to CD8+ T cells after gene gun immunization. AB - Cutaneous gene (DNA) bombardment results in substantial expression of the encoded antigen in the epidermal layer as well as detectable expression in dendritic cells (DC) in draining lymph nodes (LNs). Under these conditions, two possible modes of DC antigen presentation to naive CD8+ T cells might exist: (a) presentation directly by gene-transfected DC trafficking to local lymph nodes, and (b) cross-presentation by untransfected DC of antigen released from or associated with transfected epidermal cells. The relative contributions of these distinct modes of antigen presentation to priming for cytotoxic T cell (CTL) responses have not been clearly established. Here we show that LN cells directly expressing the DNA-encoded antigen are rare; 24 h after five abdominal skin bombardments, the number of these cells does not exceed 50-100 cells in an individual draining LN. However, over this same time period, the total number of CD11c+ DC increases more than twofold, by an average of 20,000-30,000 DC per major draining node. This augmentation is due to gold bombardment and is independent of the presence of plasmid DNA. Most antigen-bearing cells in the LNs draining the site of DNA delivery appear to be DC and can be depleted by antibodies to an intact surface protein encoded by cotransfected DNA. This finding of predominant antigen presentation by directly transfected cells is also consistent with data from studies on cotransfection with antigen and CD86 encoding DNA, showing that priming of anti-mutant influenza nucleoprotein CTLs with a single immunization is dependent upon coexpression of the DNAs encoding nucleoprotein and B7.2 in the same cells. These observations provide insight into the relative roles of direct gene expression and cross-presentation in CD8+ T cell priming using gene gun immunization, and indicate that augmentation of direct DC gene expression may enhance such priming. PMID- 9743527 TI - Antagonist peptide selects thymocytes expressing a class II major histocompatibility complex-restricted T cell receptor into the CD8 lineage. AB - CD4/CD8 lineage decision is an important event during T cell maturation in the thymus. CD8 T cell differentiation usually requires corecognition of major histocompatibility complex (MHC) class I by the T cell receptor (TCR) and CD8, whereas CD4 T cells differentiate as a consequence of MHC class II recognition by the TCR and CD4. The involvement of specific peptides in the selection of T cells expressing a particular TCR could be demonstrated so far for the CD8 lineage only. We used mice transgenic for an MHC class II-restricted TCR to investigate the role of antagonistic peptides in CD4 T cell differentiation. Interestingly, antagonists blocked the development of CD4(+) cells that normally differentiate in thymus organ culture from those mice, and they induced the generation of CD8(+) cells in thymus organ culture from mice impaired in CD4(+) cell development (invariant chain-deficient mice). These results are in line with recent observations that antagonistic signals direct differentiation into the CD8 lineage, regardless of MHC specificity. PMID- 9743528 TI - Characterization of a hemoglobin protease secreted by the pathogenic Escherichia coli strain EB1. AB - Many pathogenic bacteria can use heme compounds as a source of iron. Pathogenic Escherichia coli strains are capable of using hemoglobin as an iron source. However, the mechanism of heme acquisition from hemoglobin is not understood for this microorganism. We present the first molecular characterization of a hemoglobin protease (Hbp) from a human pathogenic E. coli strain. The enzyme also appeared to be a heme-binding protein. Affinity purification of this bifunctional protein enabled us to identify the extracellular gene product, and to clone and analyze its gene. A purification procedure developed for Hbp allowed us to perform functional studies. The protein interacted with hemoglobin, degraded it and subsequently bound the released heme. These results suggest that the protein is involved in heme acquisition by this human pathogen. Hbp belongs to the so called IgA1 protease-like proteins, as indicated by the kinetics of its membrane transfer and DNA sequence similarity. The gene of this protein appears to be located on the large pColV-K30 episome, that only has been isolated from human and animal pathogens. All these characteristics indicate that Hbp may be an important virulence factor that may play a significant role in the pathogenesis of E. coli infections. PMID- 9743529 TI - Major histocompatibility complex class I viral antigen processing in the secretory pathway defined by the trans-Golgi network protease furin. AB - Classical antigen presentation by major histocompatibility complex class I molecules involves cytosolic processing of endogenously synthesized antigens by proteasomes and translocation of processed peptides into the endoplasmic reticulum (ER) by transporters associated with antigen presentation (TAP). Alternative pathways for processing of endogenous antigens, generally involving the ER, have been suggested but not fully proved. We analyzed the potential for class I presentation of proteolytic maturation of secretory antigens in the exocytic pathway. We found that hepatitis B (HB) virus secretory core protein HBe can efficiently deliver COOH-terminally located antigenic peptides for endogenous class I loading in the absence of TAP. Antigen presentation to specific cytotoxic T lymphocytes correlates with protein maturation at the COOH terminus, since modification of maturation and transport of HBe through the secretory pathway alters antigen presentation. Both maturation and a necessary processing step occur in the Golgi or post-Golgi compartment. Antigen presentation is independent of proteasome activity, but inhibitors of the trans-Golgi network resident protease furin inhibit both HBe maturation and antigen presentation. These results define a new antigen processing pathway located in the secretory route, with a central role for proteolytic maturation mediated by the subtilisin protease family member furin as an efficient source for antigen presentation. PMID- 9743530 TI - Asymmetric cell divisions sustain long-term hematopoiesis from single-sorted human fetal liver cells. AB - Hematopoietic stem cells (HSCs) in adult marrow are believed to be derived from fetal liver precursors. To study cell kinetics involved in long-term hematopoiesis, we studied single-sorted candidate HSCs from fetal liver that were cultured in the presence of a mixture of stimulatory cytokines. After 8-10 d, the number of cells in primary cultures varied from <100 to >10,000 cells. Single cells in slow growing colonies were recloned upon reaching a 100-200 cell stage. Strikingly, the number of cells in subclones varied widely again. These results are indicative of asymmetric divisions in primitive hematopoietic cells in which proliferative potential and cell cycle properties are unevenly distributed among daughter cells. The continuous generation of functional heterogeneity among the clonal progeny of HSCs is in support of intrinsic control of stem cell fate and provides a model for the long-term maintenance of hematopoiesis in vitro and in vivo. PMID- 9743531 TI - The action of Bax and bcl-2 on T cell selection. AB - T cell development and selection in the thymus are shaped by the induction of apoptosis. However, a direct role in T cell development and selection for any of the molecules known to regulate apoptosis has remained controversial. We have studied the effect of bax and bcl-2 transgenes in recombination activation gene 1 deficient (RAG-1(-/-)) mice transgenic for the major histocompatibility complex class I-restricted F5 T cell receptor. Overexpression of a bax transgene in the thymus seriously impairs the production of mature T cells, whereas bcl-2 overexpression greatly promotes it. The effect of bax and bcl-2 overexpression on antigen-induced negative selection was studied using fetal thymic organ cultures. This analysis showed that Bcl-2 strongly inhibits negative selection, whereas Bax does not affect it. Our data directly show that Bcl-2 family members have specific roles in T cell selection and also lend support to the hypothesis that Bax and Bcl-2 can antagonize each other's action in a certain apoptosis pathway while in another they can be functionally nonreciprocal. PMID- 9743532 TI - Mast cells can secrete vascular permeability factor/ vascular endothelial cell growth factor and exhibit enhanced release after immunoglobulin E-dependent upregulation of fc epsilon receptor I expression. AB - Vascular permeability factor/vascular endothelial cell growth factor (VPF/VEGF) can both potently enhance vascular permeability and induce proliferation of vascular endothelial cells. We report here that mouse or human mast cells can produce and secrete VPF/VEGF. Mouse mast cells release VPF/VEGF upon stimulation through Fcepsilon receptor I (FcepsilonRI) or c-kit, or after challenge with the protein kinase C activator, phorbol myristate acetate, or the calcium ionophore, A23187; such mast cells can rapidly release VPF/VEGF, apparently from a preformed pool, and can then sustain release by secreting newly synthesized protein. Notably, the Fc epsilonRI-dependent secretion of VPF/VEGF by either mouse or human mast cells can be significantly increased in cells which have undergone upregulation of Fc epsilonRI surface expression by a 4-d preincubation with immunoglobulin E. These findings establish that at least one cell type, the mast cell, can be stimulated to secrete VPF/VEGF upon immunologically specific activation via a member of the multichain immune recognition receptor family. Our observations also identify a new mechanism by which mast cells can contribute to enhanced vascular permeability and/or angiogenesis, in both allergic diseases and other settings. PMID- 9743533 TI - Liver damage preferentially results from CD8(+) T cells triggered by high affinity peptide antigens. AB - Little is understood of the anatomical fate of activated T lymphocytes and the consequences they have on the tissues into which they migrate. Previous work has suggested that damaged lymphocytes migrate to the liver. This study compares class I versus class II major histocompatibility complex (MHC)-restricted ovalbumin-specific T cell antigen receptor (TCR) transgenic mice to demonstrate that after in vivo activation with antigen the emergence of CD4(-)CD8(-)B220(+) T cells occurs more frequently from a CD8(+) precursor than from CD4(+) T cells. Furthermore, this change in phenotype is conferred only by the high affinity native peptide antigen and not by lower affinity peptide variants. After activation of CD8(+) cells with only the high affinity peptide, there is also a dramatically increased number of liver lymphocytes with accompanying extensive hepatocyte damage and elevation of serum aspartate transaminase. This was not observed in mice bearing a class II MHC-restricted TCR. The findings show that CD4(-)CD8(-)B220(+) T cells preferentially derive from a CD8(+) precursor after a high intensity TCR signal. After activation, T cells can migrate to the liver and induce hepatocyte damage, and thereby serve as a model of autoimmune hepatitis. PMID- 9743534 TI - The envelope glycoprotein ectodomains determine the efficiency of CD4+ T lymphocyte depletion in simian-human immunodeficiency virus-infected macaques. AB - CD4+ T lymphocyte depletion in human immunodeficiency virus type 1 (HIV-1) infected humans underlies the development of acquired immune deficiency syndrome. Using a model in which rhesus macaques were infected with chimeric simian-human immunodeficiency viruses (SHIVs), we show that both the level of viremia and the structure of the HIV-1 envelope glycoprotein ectodomains individually contributed to the efficiency with which CD4(+) T lymphocytes were depleted. The envelope glycoproteins of recombinant SHIVs that efficiently caused loss of CD4(+) T lymphocytes exhibited increased chemokine receptor binding and membrane-fusing capacity compared with those of less pathogenic viruses. These studies identify the HIV-1 envelope glycoprotein ectodomains as determinants of CD4(+) T lymphocyte loss in vivo and provide a foundation for studying pathogenic mechanisms. PMID- 9743535 TI - Upregulation of interleukin 6 and granulocyte colony-stimulating factor receptors by transcription factor CCAAT enhancer binding protein alpha (C/EBP alpha) is critical for granulopoiesis. AB - Cytokines stimulate granulopoiesis through signaling via receptors whose expression is controlled by lineage-specific transcription factors. Previously, we demonstrated that granulocyte colony-stimulating factor (G-CSF) receptor mRNA was undetectable and granulocyte maturation blocked in CCAAT enhancer binding protein alpha (C/EBPalpha)-deficient mice. This phenotype is distinct from that of G-CSF receptor-/- mice, suggesting that other genes are likely to be adversely affected by loss of C/EBPalpha. Here we demonstrate loss of interleukin 6 (IL-6) receptor and IL-6-responsive colony-forming units (CFU-IL6) in C/EBPalpha-/- mice. The observed failure of granulopoiesis could be rescued by the addition of soluble IL-6 receptor and IL-6 or by retroviral transduction of G-CSF receptors, demonstrating that loss of both of these receptors contributes to the absolute block in granulocyte maturation observed in C/EBPalpha-deficient hematopoietic cells. The results of these and other studies suggest that additional C/EBPalpha target genes, possibly other cytokine receptors, are also important for the block in granulocyte differentiation observed in vivo in C/EBPalpha-deficient mice. PMID- 9743536 TI - APRIL, a new ligand of the tumor necrosis factor family, stimulates tumor cell growth. AB - Members of the tumor necrosis factor (TNF) family induce pleiotropic biological responses, including cell growth, differentiation, and even death. Here we describe a novel member of the TNF family designated APRIL (for a proliferation inducing ligand). Although transcripts of APRIL are of low abundance in normal tissues, high levels of mRNA are detected in transformed cell lines, and in human cancers of colon, thyroid, and lymphoid tissues in vivo. The addition of recombinant APRIL to various tumor cells stimulates their proliferation. Moreover, APRIL-transfected NIH-3T3 cells show an increased rate of tumor growth in nude mice compared with the parental cell line. These findings suggest that APRIL may be implicated in the regulation of tumor cell growth. PMID- 9743537 TI - A signal transducer and activator of transcription (Stat)4-independent pathway for the development of T helper type 1 cells. AB - The differentiation of T helper (Th) cells is regulated by members of the signal transducer and activator of transcription (STAT) family of signaling molecules. We have generated mice lacking both Stat4 and Stat6 to examine the ability of Th cells to develop in the absence of these two transcription factors. Stat4, Stat6( /-) lymphocytes fail to differentiate into interleukin (IL)-4-secreting Th2 cells. However, in contrast to Stat4(-/-) lymphocytes, T cells from Stat4, Stat6( /-) mice produce significant amounts of interferon (IFN)-gamma when activated in vitro. Although Stat4, Stat6(-/-) lymphocytes produce less IFN-gamma than IL-12 stimulated control lymphocytes, equivalent numbers of IFN-gamma-secreting cells can be generated from cultures of Stat4, Stat6(-/-) lymphocytes activated under neutral conditions and control lymphocytes activated under Th1 cell-promoting conditions. Moreover, Stat4, Stat6(-/-) mice are able to mount an in vivo Th1 cell-mediated delayed-type hypersensitivity response. These results support a model of Th cell differentiation in which the generation of Th2 cells requires Stat6, whereas a Stat4-independent pathway exists for the development of Th1 cells. PMID- 9743538 TI - Plasmodium falciparum produces prostaglandins that are pyrogenic, somnogenic, and immunosuppressive substances in humans. AB - Plasmodium falciparum causes the most severe form of human malaria, which kills approximately 1.5-2.7 million people every year, but the molecular mechanisms underlying the clinical symptoms and the host-parasite interaction remain unclear. We show here that P. falciparum produces prostaglandins (PGs) D2, E2, and F2alpha. After incubation with 1 mM arachidonic acid (AA), cell homogenates of P. falciparum produced PGs as determined by enzyme immunoassay and gas chromatography-selected ion monitoring. PG production in the parasite homogenate was not affected by the nonsteroidal antiinflammatory drugs aspirin and indomethacin, and was partially heat resistant, whereas PG biosynthesis by mammalian cyclooxygenase was completely inhibited by these chemicals and by heat treatment. Addition of AA to the parasite cell culture markedly increased an ability of the parasite cell homogenate to produce PGs and of parasitized red blood cells to accumulate PGs in the culture medium. PGD2 and PGE2 accumulated in the culture medium at the stages of trophozoites and schizonts more actively than at the ring stage. These findings are the first evidence of the direct involvement of a malaria parasite in the generation of substances that are pyrogenic and injurious to the host defenses. We will discuss a possible contribution of the parasite-produced PGs to pathogenesis and host-parasite interaction of P. falciparum. PMID- 9743540 TI - Exon screening of the genes encoding the beta- and gamma-subunits of cone transducin in patients with inherited retinal disease. AB - PURPOSE: To screen the exons of the genes encoding the beta3-subunit (GNB3) and gammac-subunit (GNGT2) of cone transducin for mutations in a large number of unrelated patients with various forms of inherited retinal disease including cone dystrophy, cone-rod dystrophy and macular dystrophy. METHODS: Exons of the two genes were screened for mutations by denaturing gradient gel electrophoresis (DGGE) and/or single strand conformation polymorphism electrophoresis (SSCP); any variants were sequenced directly. RESULTS: Although many sequence variants were found in both genes, none could be associated with disease. Additionally, the gene structure and sequence of the coding exons of GNB3 were determined and compared with those of the dog homolog. Both human and canine GNB3 have nine coding exons and their two predicted amino acid sequences have 97% identity. CONCLUSIONS: The results indicate that GNB3 and GNGT2 are unlikely sites of mutations responsible for inherited retinal degenerations that predominantly effect cone-mediated function (cone and cone-rod dystrophies) or have a predilection for disease in the macula (macular dystrophies). PMID- 9743539 TI - High frequency of skin-homing melanocyte-specific cytotoxic T lymphocytes in autoimmune vitiligo. AB - Vitiligo is an autoimmune condition characterized by loss of epidermal melanocytes. Using tetrameric complexes of human histocompatibility leukocyte antigen (HLA) class I to identify antigen-specific T cells ex vivo, we observed high frequencies of circulating MelanA-specific, A*0201-restricted cytotoxic T lymphocytes (A2-MelanA tetramer+ CTLs) in seven of nine HLA-A*0201-positive individuals with vitiligo. Isolated A2-MelanA tetramer+ CTLs were able to lyse A*0201-matched melanoma cells in vitro and their frequency ex vivo correlated with extent of disease. In contrast, no A2-MelanA tetramer+ CTL could be identified ex vivo in all four A*0201-negative vitiligo patients or five of six A*0201-positive asymptomatic controls. Finally, we observed that the A2-MelanA tetramer+ CTLs isolated from vitiligo patients expressed high levels of the skin homing receptor, cutaneous lymphocyte-associated antigen, which was absent from the CTLs seen in the single A*0201-positive normal control. These data are consistent with a role of skin-homing autoreactive melanocyte-specific CTLs in causing the destruction of melanocytes seen in autoimmune vitiligo. Lack of homing receptors on the surface of autoreactive CTLs could be a mechanism to control peripheral tolerance in vivo. PMID- 9743541 TI - Up-regulation of osteonectin/SPARC in age-related cataractous human lens epithelia. AB - PURPOSE: To characterize gene expression patterns between epithelia isolated from cataractous and normal human lenses. METHODS: Reverse transcriptase differential display was used to identify differential expression between cataractous and normal epithelia. RT-PCR was used to compare pooled and individual RNA samples. RESULTS: One transcript, up-regulated in cataractous as compared to normal epithelia, was identified as osteonectin which is also known as SPARC (secreted acidic protein rich in cysteines). RT-PCR confirmed over-expression of this RNA. High levels of osteonectin mRNA were also detected in six individual epithelia dissected from cataractous lenses. CONCLUSIONS: The present study provides evidence for up-regulation of osteonectin in human age-related cataract and suggests that osteonectin, a protein involved in cell-cycle control, extracellular matrix and Ca++ binding, plays an important role in human lens homeostasis and may be involved in processes leading to lens opacity. PMID- 9743542 TI - Impaired speech perception in poor readers: evidence from hearing and speech reading. AB - The performance of 14 poor readers on an audiovisual speech perception task was compared with 14 normal subjects matched on chronological age (CA) and 14 subjects matched on reading age (RA). The task consisted of identifying synthetic speech varying in place of articulation on an acoustic 9-point continuum between /ba/ and /da/ (Massaro & Cohen, 1983). The acoustic speech events were factorially combined with the visual articulation of /ba/, /da/, or none. In addition, the visual-only articulation of /ba/ or /da/ was presented. The results showed that (1) poor readers were less categorical than CA and RA in the identification of the auditory speech events and (2) that they were worse in speech reading. This convergence between the deficits clearly suggests that the auditory speech processing difficulty of poor readers is speech specific and relates to the processing of phonological information. PMID- 9743543 TI - MRI asymmetries of Broca's area: the pars triangularis and pars opercularis. AB - Broca's area, which includes the pars triangularis (PTR) and pars opercularis (POP), is a neuroanatomic region important in speech-language production. Previous data demonstrated that PTR asymmetries are highly correlated with language dominance determined by selective hemispheric anesthesia or Wada testing, suggesting that asymmetries of the PTR may, in part, predict language dominance. The POP, however, has not been measured on volumetric magnetic resonance imaging (MRI), and therefore, it is unclear whether morphological asymmetries of the POP exist, and whether these asymmetries differ in right- and left-handers. The purpose of this study was to determine if measurable asymmetries of the POP exist on MRI, and whether the direction of the asymmetries differ in right- and left-handers. The PTR and POP were measured on volumetric MRI scans of 16 right-handers and 16 left-handers matched for age and gender. There was a significant leftward asymmetry of the PTR in right- and left-handers, although the asymmetry was reduced in the left-handers. In contrast, there was a leftward asymmetry of the POP in right-handers, and a rightward asymmetry in the left-handers. Handedness, derived from a handedness inventory, was positively correlated with POP asymmetry. PMID- 9743544 TI - Sentence comprehension deficits in Alzheimer's disease: a comparison of off-line vs. on-line sentence processing. AB - Two studies explored whether sentence comprehension impairments in Alzheimer's disease (AD) are due to deficits in syntactic processing or memory. Study 1 used a picture-pointing sentence comprehension task to measure the final outcome of comprehension in an off-line fashion. It showed the comprehension of 30 patients with AD to be impaired, but suggested that the deficits could not be attributed solely to syntactic impairments. Study 2 investigated the effects of memory on sentence comprehension by comparing off-line (grammaticality judgment) with on line (cross-modal naming) language processing in 11 AD and 9 control subjects. The results revealed impaired performance in the off-line task but normal performance in the on-line task using the same sentences. Performance on the off line task correlated with independent measures of verbal working memory. These data are used to argue that sentence comprehension impairments are related to verbal working memory deficits in AD. PMID- 9743545 TI - Neurons in human temporal cortex active with verbal associative learning. AB - In neuronal activity recorded from human middle temporal gyrus during learning of associations between word pairs, a population was identified that had greater activity for associations that were learned rapidly during initial encoding compared to those learned slowly or not at all by an individual subject. This population can be separated from other neurons by the combination of inhibition during word reading when no learning is required and excitation during recent memory for words. These neurons are present in both hemispheres, predominately in deeper layers of cortex. During initial encoding, the increased activity appears at presentation of all word pairs but persists for several seconds only for the rapidly learned pairs, likely reflecting rehersal of items being learned. Human associative learning is related to activity of this specific population of "association" neurons, identified here for the first time. PMID- 9743546 TI - Understanding disorders in agrammatic patients: capacity or structural deficits? AB - Several hypotheses have been advanced in recent years to understand difficulties in agrammatic patients. Some of them are of a structural kind, as the deficiency is said to lie in some of the linguistic system components. Others are of a functional type, as it is stated that the problem of these patients lies in the loss of processing capacity. Using the existence of a syntactic type of structure in Spanish, that active sentences do not follow the canonical S-V-O order, we will try to prove in this article whether agrammatic patients' problems are due to memory span loss or to one of the syntactic process mechanisms. To this end, the performances of three groups of patients are contrasted. Agrammatic, anomic, and normal Spanish speakers are given several tasks of sentence-picture matching and tests of memory span. Results show that agrammatic patients have specific difficulties processing certain syntactical structures; however, their memory deficiencies are not more pronounced than in other patients. It can be concluded, therefore, that the deficiencies of agrammatic patients are of a structural character rather than due to memory span loss. PMID- 9743547 TI - Distributed versus localist representations: evidence from a study of item consistency in a case of classical anomia. AB - Models of word production and comprehension can be split into two broad classes: localist and distributed. In localist architectures each word within the lexicon is represented by a single unit. The distributed approach, on the other hand, encodes each lexical item as a pattern of activation across a set of shared units. If we assume that the localist representations are more than a convenient shorthand for distributed representations at the neuroanatomical level, it should be possible to find patients who, after brain injury, have lost specific words from their premorbid vocabulary. Following a closed head injury, JS had severe word-finding difficulties with no measurable semantic impairment nor did he make phonological errors in naming. Cueing with an initial phoneme proved relatively ineffective. JS showed a high degree of item consistency across three administrations of two tests of naming to confrontation. This consistency could not be predicted from a linear combination of psycholinguistic variables but the distribution fitted a stochastic model in which it is assumed that a proportion of items have become consistently unavailable. Further evidence is presented which suggests that these items are not, in fact, lost but rather have a very low probability of retrieval. Given phonemic cueing of sufficient length, or delayed repetition priming from a written word, the consistently unnamed items were produced by JS. Additional data is reported which seems to support a distributed model of speech production. JS's naming accuracy for one set of pictures was found to predict his performance on a second set of items only when the names of the pictures were both semantically and phonologically related (e.g., cat-rat). There was no association for pairs of pictures if they were only semantically (e.g., cat-dog) or phonologically related (e.g., cat-cap). It is argued that JS's data are best described in terms of a graded, non-linear, distributed model of speech production. PMID- 9743548 TI - Locus of functional impairment in the production of speech rhythm after brain damage: a preliminary study. AB - This acoustic-perceptual, multiple-case study of the Rhythm Rule (RR) in English, a phonological phenomenon whereby adjacent stresses are adjusted to avoid "stress clash" (e.g., thirTEEN vs THIRteen MEN), was undertaken to identify the locus of functional impairments in speech prosody in different aphasic syndromes. Subjects included two left brain-damaged aphasic patients (1 fluent, 1 nonfluent), one right brain-damaged nonaphasic patient, and one nonneurological control. They were instructed to read sentences including experimental (bisyllabic "double stressed" words) and matching control (bisyllabic "initial-stressed" words) phrases of increasing length. Results of acoustic and perceptual measures indicated that rhythmic disturbances associated with the RR emerged regardless of lesion site. The locus of functional impairment was isolated to phonetic implementation of the RR, as opposed to either loss of word-level stress or loss of the RR. Findings suggest that neural substrates of speech prosody are broadly distributed in the left and right cerebral hemispheres. PMID- 9743549 TI - Right hemisphere activation during indirect semantic priming: evidence from event related potentials. AB - Healthy subjects performed a lexical decision task in a semantic priming paradigm while event-related potentials (ERPs) were recorded from 64 channels. Semantic distance between prime and target was varied by including directly, indirectly, and nonrelated word pairs. At centro-parietal electrodes an N400 to nonrelated pairs was elicited bilaterally which was sensitive only to direct, but not to indirect semantic priming. These N400 priming effects were mirrored by the RT data. At inferior fronto-temporal sites directly related words showed ERP priming effects over both hemispheres. However, indirectly related words only elicited ERP priming effects over the right hemisphere. These results support the hypothesis that the right hemisphere semantic system is involved in processing of remote semantic information. PMID- 9743550 TI - Language development in children with simple-partial left-hemisphere epilepsy. AB - The nature of cerebral involvement in the acquisition of language was addressed in this longitudinal study of children with an early diagnosis of epilepsy with simple-partial seizures (SPE) and with epileptogenic foci localized in the left frontal (LF) lobe. Yearly evaluations of six SPE-LF children on tests of linguistic comprehension (pointing, understanding of narrative, and understanding of prepositions) and production (repetition, lexical diversity, and grammatical production) were carried out between the ages of 3 and 8 years and compared to those of large samples of control children on the same tasks and at each age level. Linguistic production of all children were transcribed, coded, and analyzed using the Child Language Data Exchange System (MacWhinney & Snow, 1991). Individual evolution trajectories revealed that SPE-LF children showed a clear dissociation in linguistic performance between comprehension and production. Linguistic comprehension gradually improved to reach normal performance levels by age 7 while linguistic production, even at later stages, remained quite poor. This dissociation in the development of linguistic performance in SPE-LF children suggests a complex interplay between brain maturation dynamics and dysfunction modulating the succession of stages in language development. The observed persistent deficits in specific aspects of linguistic performance argue for an early involvement of the anterior areas of the left cerebral hemisphere in the production of language. PMID- 9743552 TI - Visualizing memory phenotype development after in vitro stimulation of CD4(+) T cells. AB - Stimulation of naive CD4 cells by specific antigen results in proliferation and changes in cell surface marker expression as the cells differentiate into effector and memory cells. Several of the marker changes (e.g., differences in CD45RB, CD44, and L-selectin levels) appear to be relatively stable and permit the identification of memory T cells. In this study, we examined the acquisition of memory markers after the initial stimulation of naive T cells. CD4(+) T cells from DO11.10 TCR transgenic mice were labeled with the fluorescent dye carboxyfluorescein diacetate succinimidyl ester (CFSE) and were stimulated with specific antigen (OVA323-339). Specific activation was observed, as CFSE associated fluorescence was reduced twofold with each division of DO11.10 clonotype-bearing cells. Phenotypic changes could also be observed as the cells differentiated into effector/memory cells. However, individual surface markers exhibited a varied relationship to cell division. Although changes in some markers (L-selectin) occurred independently of cell division, changes in other markers were either strictly related to cell division (CD45RB) or were a prerequisite to cell division (CD4, CD44). PMID- 9743553 TI - A carbocyclic nucleoside analogue is a TNF-alpha inhibitor with immunosuppressive action: role of prostaglandin E2 and protein kinase C and comparison with pentoxifylline. AB - Tumor necrosis factor-alpha (TNF-alpha) is associated with several acute and chronic inflammatory conditions. New therapies directed at inhibiting TNF-alpha will be important in treating pathological processes mediated by TNF-alpha. In this study, we studied and compared the effect of the carbocyclic nucleoside analogue (9-[(1R, 3R)-trans-cyclopentan-3-ol] adenine) with pentoxifylline on modulating TNF-alpha production. The carbocyclic nucleoside analogue inhibited TNF-alpha production in a dose-dependent manner (1 microM-1 mM) by stimulated peripheral blood mononuclear cells and cell lines of both monocyte (THP-1) and T lymphocyte phenotypes (CEM x 174). The drug potently inhibited TNF production in cells stimulated by endotoxin, the superantigen (staphylococci enterotoxin A), the mitogen (phytohemagglutinin), and the protein kinase C activator (phorbol myristate acetate) with ED50 ranging from 5 to 30 microM. At moderate concentrations, the carbocyclic nucleoside analogue inhibited superantigen (ED50 = 300 microM) and alloantigen (mixed lymphocyte reaction) T cell proliferative responses (ED50 = 150 microM). The involvement of protein kinase C and prostaglandin E2 (PGE2), mediators that regulate TNF-alpha production, was also investigated. Unlike PTX, the nucleoside analogue did not upregulate PGE2 production. The inhibition of TNF-alpha production appeared to be mediated at least partly by PKC, since the nucleoside analogue caused suppression of PKC activity in stimulated cells. The results show that the carbocyclic nucleoside analogue is a TNF-alpha inhibitor that may be appropriate in the therapy of TNF alpha-associated complications. The suppressive properties of the carbocyclic nucleoside analogue on antigen and alloantigen (mixed lymphocyte reaction) responses may be appropriate in disease conditions in which inhibiting both TNF alpha and T-cell reactivity is desirable. PMID- 9743554 TI - Mice deficient in fas ligand (gld) or fas (lpr) show few alterations in granulopoiesis. AB - Evidence exists that the life span of mature, circulating neutrophils is influenced by apoptosis induced by interactions between Fas ligand (FasL) and Fas (CD95). However, the role of FasL/Fas-mediated apoptosis in granulopoiesis has not been explored. The present study assessed differences in granulopoiesis between normal (B6) mice and mice carrying mutations in the genes for FasL (B6/gld) or Fas (B6/lpr). Granulopoiesis was examined by quantitating mature granulocytes in the blood, committed myeloid progenitor cells (or colony-forming units; CFU) in the bone marrow (BM), and granulocyte lineage cells in the BM. The present study also characterized through flow cytometry the ability of GR-1(+) granulocyte lineage cells from B6, B6/gld, and B6/lpr mice to undergo spontaneous apoptosis in vitro. In comparison to B6 mice, B6/gld mice (but not B6/lpr mice) showed small, but significant increases in the number and percentage of blood granulocytes and in the number of myeloid CFU. However, the number and percentage of GR-1(+) granulocyte lineage cells in the BM were similar in the three strains. The rate of spontaneous apoptosis of GR-1(+) granulocyte lineage cells also did not differ between B6, B6/gld, and B6/lpr mice. In B6 and B6/gld mice, Fas expression on granulocyte lineage cells was downregulated in conjunction with a decrease in forward-angle light scatter (fsc) and externalization of phosphatidylserine (PS), two measures of apoptosis. These results suggest that FasL-Fas interactions play only a minor role in modulating numbers of committed myeloid progenitor cells and the size of the peripheral pool of mature granulocytes. Interactions between FasL and Fas do not influence the size of the BM pool of granulocyte lineage cells or the ability of those cells to undergo spontaneous apoptosis. PMID- 9743555 TI - Receptor-mediated activation of murine peritoneal macrophages by antithrombin III acts as a costimulatory signal for nitric oxide synthesis. AB - We evaluated the effect of antithrombin III (ATIII), a serine protease inhibitor (SERPIN), on induction of nitric oxide (NO) synthesis in murine peritoneal macrophages. Incubation of macrophages with ATIII plus interferon-gamma (IFN gamma) but not ATIII alone induced nitrite accumulation (a metabolite of NO) in a dose-dependent manner. Expression of the inducible nitric oxide synthase isoform was confirmed by Western blot. NO synthesis was inhibited by NG-monomethyl-l arginine, by complexing ATIII with thrombin or by rabbit anti-human ATIII antiserum. Addition of polymyxin B to macrophage cultures failed to inhibit ATIII/IFN-gamma-induced NO synthesis, excluding lipopolysaccharide contamination. 125I-ATIII bound to macrophages in a dose-dependent, specific, and saturable manner, with a Km of approximately 7.1 nM. Our results demonstrate that ATIII, but not ATIII/thrombin complex, acts to costimulate macrophage activation and NO synthesis via a novel receptor mediated mechanism, which may indicate a role for SERPINs in macrophage activation. PMID- 9743556 TI - Inhibition of IgG2ab production by Ig allotype-specific T cells can Be mediated without T-B cell contact. AB - The growth of IgG2ab-producing CB101 myeloma cells, subcutaneously or intraperitoneally inoculated into histocompatible BALB/c Igha mice sensitized against this Ig allotype, was delayed by 2-4 weeks compared to normal mice. While IgG2ab production was detected in the sera of 75-100% of normal mice, it was irreversibly inhibited in 100% of sensitized mice. IgG2ab suppression (IgG2ab sup) was also systematically obtained in sensitized but not normal recipients, implanted ip with a 0.1-micrometer-pore diffusion chamber (DC) containing CB101 cells. This time, the specific IgG2ab sup was reversible in vitro in the absence of anti-IgG2ab T cells. Adoptive transfer, of unfractionated or T but not B splenocytes from their sensitized counterparts into normal mice, 1 day before DC implantation, induced IgG2ab sup as well. These results indicate that, in these experimental circumstances, IgG2ab sup can also be mediated by diffusible suppressive factors produced by the effector T cells, without direct T-B-cell contact. PMID- 9743557 TI - T cell recognition of flanking residues of murine invariant chain-derived CLIP peptide bound to MHC class II. AB - The major site of interaction between MHC class II molecules and invariant chain has been mapped to occupancy of the class II peptide-binding site by the CLIP region of invariant chain. CLIP is also seen as a degradation product of invariant chain and can be found in association with class II as a processing intermediate. Here we analyzed the relative contribution of single amino acids in the murine CLIP (86-102) peptide for binding to I-Ab and I-Ad and for recognition by a CLIP-specific T cell hybridoma. Interestingly, the I-Ab-restricted murine T cell hybridoma that recognizes murine CLIP peptide (86-102) is dependent on Met 102 for activation. This amino acid is outside of the core binding region and in the CLIP/DR3 crystal structure extends outside of the class II peptide-binding site. These data suggest that a T cell epitope presented on CLIP/class II complexes can be located predominantly in flanking residues that extend out of the peptide binding groove of class II. PMID- 9743558 TI - T-Helper 1 and T-helper 2 cytokine responses in gut-associated lymphoid tissue following enteric reovirus infection. AB - Enteric infection of mice with reovirus serotype 1 elicits antibody and cytotoxic T-lymphocytes in gut-associated lymphoid tissue (GALT). This led to the hypothesis that T-helper 1 (Th1) and T-helper 2 (Th2) responses develop in GALT. Reverse transcriptase-polymerase chain reactions on RNA from Peyer's patches (PP), intraepithelial lymphocytes (IEL), and lamina propria (LP) lymphocytes demonstrated that interferon (IFN)-gamma message was increased in PP and IEL, but not in LP following infection. No increase in mRNA for interleukin (IL)-4, IL-5, or IL-6 was detected. IFN-gamma, IL-5, and IL-6 were produced in in vitro cultures of PP 4-10 days postinfection. PP and spleen lymphocytes from infected mice produced IFN-gamma, but no IL-5 following in vitro restimulation. Infection also induced production of mRNA for the beta2 chain of the IL-12 receptor in PP. We conclude that reovirus induces robust Th1 and weak Th2 cell responses in GALT. PMID- 9743559 TI - IL-6-deficient mice form granulomas in murine schistosomiasis that exhibit an altered B cell response. AB - IL-6 can play an important role in various biological activities. Using IL-6 deficient, 129 x C57BL/6 mice and normal littermate controls, we studied the role of IL-6 in granulomas of mice infected with schistosomiasis mansoni. Granulomas from IL-6(+/+) mice produced large quantities of IL-6, derived from T, B, and myeloid cells. Yet, IL-6 mutant mice generated normal-appearing granulomas of appropriate size. Multiple-parameter flow cytometric analysis of dispersed granuloma cells revealed no substantial differences. Granuloma cells and splenocytes were cultured in vitro to measure cytokine and immunoglobulin production. Compared to control cells, IL-6(-/-) granuloma cells secreted more IL 4, IL-5, and IL-10. However, splenocytes secreted cytokines comparably. In the IL 6(-/-) state, the granuloma cells released less IgE and substantially more IgM, although IgG1, IgG2a, and IgA secretion remained normal. ELISPOT assay showed that dispersed granuloma cells from IL-6-deficient animals had substantially more IgM-secreting B cells. Thus, schistosome granulomas make IL-6 that is not essential for most aspects of granuloma development. However, IL-6 deficiency results in some disturbance of granuloma cytokine and immunoglobulin expression. PMID- 9743560 TI - The T cell repertoire primed by antiviral vaccination is influenced by self tolerance. AB - Vaccination can elicit CD8(+) cytotoxic T lymphocytes (CTL) that recognize peptides presented by class I MHC molecules. Relatively little is known, however, about the genetic factors that shape the repertoire of T cell clonotypes responding to any given epitope. We report here that H-2(b) mice immunized with a plasmid DNA vaccine or vaccinia virus encoding for HIV-1SF2p55gag elicit CD8(+) CTL against the H-2Db-restricted immunodominant epitope (pgagb). This response involved three different T cell populations based on their recognition of alloantigens: one that cross-reacted with the alloantigen H-2Ld, one that cross reacted with H-2Kd, and one that did not cross-react with either H-2(d) or H-2(k) molecules. Using the TAP-deficient cell line T2-Ld, we showed that pgagb-specific CTL cross-react with H-2Ld and a yet unidentified self-peptide. In mice expressing H-2(b) and H-2(d) allotypes, we investigated whether tolerance to H 2(d) influenced the HIVp55gag-specific CTL repertoire as a consequence of thymic deletion of the cross-reactive CTL repertoire. In (H-2(dxb))F1 mice heterogygosity at the MHC-I level prevented maturation of some but not all TCR combinations specific for H-2Db+pgagb, illustrating the concept that self tolerance can influence the repertoire of antiviral T cells. PMID- 9743562 TI - Degeneration of spared axons following partial white matter lesion: implications for optic nerve neuropathies. AB - Neuroprotective therapy is a relatively new development in the approach to the treatment of acute and chronic brain damage. Though initially viewed in the framework of acute CNS injuries, the concept was recently extended to include chronic injuries, in which at any given time there are some neurons in an acute phase of degeneration coexisting with others that are healthy, marginally damaged, or dead. The healthy neurons and those that are only marginally damaged are the potential targets for neuroprotection. For the development of neuroprotective therapies, it is essential to employ an animal model in which the damage resulting from secondary degeneration can be quantitatively distinguished from primary degeneration. This is of particular relevance when the site of the damage is in the white matter (nerve fibers) rather than in the gray matter (cell bodies). In the present work we reexamine the concepts of secondary degeneration and neuroprotection in white matter lesions. Using a partial crush injury of the adult rat optic nerve as a model, we were able to assess both primary and secondary nerve damage. We show that neurons whose axons were not damaged or only marginally damaged after an acute insult will eventually degenerate as a consequence of their existence in the degenerative environment produced by the injury. This secondary degeneration does not occur in all of the neurons at once, but affects them in a stepwise fashion related to the severity of the damage inflicted. These findings, which may be applicable to the progression of acute or chronic neuropathy, imply that neuroprotective therapy may have a beneficial effect even if there is a time lag between injury and treatment. PMID- 9743563 TI - Multivariate analysis of laminar patterns of neurodegeneration in posterior cingulate cortex in Alzheimer's disease. AB - Posterior cingulate cortex is the site of earliest reductions in glucose metabolism and qualitatively different laminar patterns of neurodegeneration in Alzheimer's disease (AD). This study used multivariate analyses of area 23 in 72 cases of definite AD to assess relationships between laminar patterns of neurodegeneration, neurofibrillary tangle (NFT) and senile plaque (SP) densities, age of disease onset and duration, and apolipoprotein E (ApoE) genotype. No age related changes in neurons occurred over four decades in 17 controls and regression analysis of all AD cases showed no relationships between neuron, SP, and tau-immunoreactive NFT densities. Principal components analysis of neurons in layers III-Va and eigenvector projections showed five subgroups. The subgroups were independent because each had a full range of disease durations and qualitatively different laminar patterns in degeneration suggested disease subtypes (ST). Cases with most severe neuron losses (STSevere) had an early onset, most SP, and highest proportion of ApoE epsilon4 homozygotes. Changes in the distribution of NFT were similar over disease course in two subtypes and NFT did not account for most neurodegeneration. In STII-V with moderate neuron loss in most layers, cases with no NFT had a disease duration of 3.5 +/- 0.9 years (mean +/- SEM), those with most in layers IIIc or Va had a duration of 7.3 +/- 1 years, and those with most in layers II-IIIab had a duration of 12.1 +/- 1 years. In STSevere, cases with highest NFT densities in layers II-IIIab also were late stage. Finally, epsilon4 homozygotes were most frequent in STSevere, but four statistical tests showed that this risk is not directly involved in neurodegeneration. In conclusion, multivariate pattern recognition shows that AD is composed of independent neuropathological subtypes and NFT in area 23 do not account for most neuron losses. PMID- 9743564 TI - Nerve growth factor (NGF) and diabetic neuropathy in the rat: morphological investigations of the sural nerve, dorsal root ganglion, and spinal cord. AB - A number of functions for nerve growth factor (NGF) have been described over the past years, including its role for neuronal function and regeneration during toxic or metabolic neuropathies. In order to further assess the effects of NGF on the somatosensory system in diabetic neuropathy, the sural nerve, dorsal root ganglia (DRG), and dorsal horn of the spinal cord were investigated by morphological and quantitative methods in rats after 12 weeks of uncontrolled streptozotocin-induced diabetes mellitus. The results from our study suggest a twofold effect of NGF: (1) In sural nerve treatment with NGF (0.1 or 0.5 mg/kg) for 12 weeks was able to reverse distinct diabetes-related alterations in myelinated nerve fiber morphology, such as myelin thickness. These changes occurred in the entire myelinated population of sensory nerves and were not restricted to nociceptive nerve fibers. (2) The NGF effect on neurotransmitters of the sensory, nociceptive system was reflected by increased CGRP and substance P content in the DRG and in the dorsal horn of the spinal cord. No change of trkA receptor immunostaining was seen in DRGs of diabetic rats; however, a reduction of trkA immunoreactivity of DRG neurons was noted after long-term NGF treatment of healthy controls. The data demonstrate that NGF regulates a number of neuronal parameters along peripheral and central parts of the somatosensory pathway in the adult. This neurotrophic support may be essential for inducing functionally significant regenerative mechanisms in diabetic neuropathy. PMID- 9743565 TI - Evidence for synaptic apoptosis. AB - Increasing evidence indicates that neurons die by apoptosis, an active form of cell death involving a relatively stereotyped series of biochemical changes that culminate in nuclear fragmentation, in many different developmental and pathophysiological settings. In contrast to most other cell types, neurons have elaborate morphologies with complex neuritic arbors that often extend great distances from the cell body. Neuronal death signals are likely to be activated at remote synaptic sites and, indeed, overactivation of glutamate receptors and underactivation of trophic factor receptors are implicated in neurodegenerative disorders. We now report that biochemical changes consistent with apoptosis are engaged locally in synapses. Exposure of cortical synaptosomes to staurosporine and Fe2+ resulted in loss of membrane phospholipid asymmetry, caspase activation, and mitochondrial alterations (membrane depolarization, calcium overload, and oxyradical accumulation) characteristic of apoptosis. The caspase inhibitor zVAD fmk prevented mitochondrial membrane depolarization in synaptosomes. Studies of the effects of cytosolic extracts from synaptosomes exposed to apoptotic insults, on isolated nuclei, showed that signals capable of inducing nuclear apoptosis are generated locally in synapses. Exposure of cultured hippocampal neurons to staurosporine and glutamate resulted in caspase activation and mitochondrial membrane depolarization in dendrites, and zVAD-fmk prevented the membrane depolarization. Glutamate-induced increases in caspase activity were first observed in dendrites and later in the cell body, and focal application of glutamate to individual dendrites resulted in local activation of caspases. Collectively, the data demonstrate that apoptotic biochemical cascades can be activated locally in synapses and dendrites and suggest a role for such local apoptotic signals in synapse loss and neuronal death in neurodegenerative disorders that involve excessive activation of glutamate receptors. PMID- 9743566 TI - Collagen containing neurotrophin-3 (NT-3) attracts regrowing injured corticospinal axons in the adult rat spinal cord and promotes partial functional recovery. AB - During development, neurotrophic factors play an important role in the guidance and outgrowth of axons. Our working hypothesis is that neurotrophic factors involved in the development of axons of a particular CNS tract are among the most promising candidates for stimulating and directing the regrowth of fibers of this tract in the lesioned adult animal. The neurotrophin NT-3 is known to be involved in the target selection of outgrowing corticospinal tract (CST) fibers. We studied the capacity of locally applied NT-3 to stimulate and direct the regrowth of axons of the CST in the lesioned adult rat spinal cord. We also studied the effect of NT-3 application on the functional recovery of rats after spinal cord injury, using the gridwalk test. NT-3 was applied at the site of the lesion dissolved into rat tail collagen type I. Four weeks after spinal cord injury and collagen implantation, significantly more CST fibers had regrown into the collagen matrix containing NT-3 (22 +/- 6%, mean +/- SEM) than into the control collagen matrix without NT-3 (7 +/- 2%). No CST fibers grew into areas caudal to the collagen implant. Despite the absence of regrowth of corticospinal axons into host tissue caudal to the lesion area, functional recovery was observed in rats with NT-3 containing collagen implants. PMID- 9743567 TI - White matter astrocytes produce hepatocyte growth factor activator inhibitor in human brain tissues. AB - Hepatocyte growth factor (HGF) is a potent mitogen for mature hepatocytes and also has multifunctional effects on some other cells in various organs. A HGF activator (HGFA) has been identified as a key enzyme that regulates the activity of HGF in vivo. Our previous studies have shown that brain astrocytes produce both proteins. Recently, HGFA inhibitor-1 (HAI-1), a novel Kunitz-type serine protease inhibitor, has been isolated. We examined HAI-1 immunolabeling in the brains of neurologically normal persons and patients with Alzheimer's disease (AD) and cerebral infarction. Furthermore, we identified the expression of the mRNA for HAI-1 by in situ hybridization histochemistry. The HAI-1 antibody stained astrocytes in the white matter of all brain tissues and was present in plasma. In AD, the intensity of HAI-1 immunolabeling was less than in the other cases. Expression of the mRNA for HAI-1 was also seen in astrocytes. The intensity of the signal for HAI-1 mRNA was similar in AD and normal control brains. These results suggest that, in human brain, secreted pro-HGF from astrocytes may be activated by HGFA and inhibited by HAI-1 on or near the astrocytic cell surface and that rapid HAI-1 consumption may occur in the white matter in AD. PMID- 9743568 TI - Rabies virus entry into endosomes in IMR-32 human neuroblastoma cells. AB - Early events in rabies virus entry into cultured IMR-32 human neuroblastoma cells were investigated. After adsorption of rabies virus to the cell surface in the cold and warming to 37 degrees C in the presence of tracers for early endosomes, rabies virus and tracers were localized by immunofluorescence microscopy. After 5 min, rabies virus colocalized with Lucifer Yellow, Texas Red-dextran, rhodamine wheat germ agglutinin, and transferrin receptor in puncta in the cell body, neurites, and nerve terminals. Rabies virus did not colocalize with lysosomal glycoprotein. An acidotropic probe revealed that some of the virus-containing puncta were acidified. Rabies virus also colocalized with synapsin I, a synaptic vesicle marker, in swellings along processes, indicating some virus enters nerve terminals. Electron microscopy revealed the presence of rabies virus within irregular membrane compartments located near the cell surface in the cell body and neurites. The membrane of the virus particle was often continuous with that of the vacuole. It is concluded that rabies virus enters IMR-32 neuroblastoma cells by adsorptive endocytosis and that, shortly after entry, rabies virus is located within and fuses with acidic endosomes. PMID- 9743569 TI - Metabolic compromise with systemic 3-nitropropionic acid produces striatal apoptosis in Sprague-Dawley rats but not in BALB/c ByJ mice. AB - Metabolic compromise with systemic 3-nitropropionic acid (3-NP) results in the degeneration of striatal cells, mimicking the pathology of Huntington's disease (HD). Here we show that 10-week- and 8-month-old BALB/c ByJ mice show an unexpected striatal resilience to single and multiple systemic injections of 3 NP, while Sprague-Dawley rats are vulnerable, albeit in a variable manner. Identification of lesions was made by staining of DNA fragmentation with terminal deoxytransferase-mediated dUTP-biotin nick-end labeling (TUNEL) and hematoxylin/eosin, 1-10 days after injection. Quantitative imaging of histochemistry for succinate dehydrogenase (SDH) activity, the target of 3-NP inhibition, revealed that vulnerable rats reached maximal inhibition in brain at 1 day after 3-NP, whereas mice and resilient rats took 7 days to reach maximal inhibition. All groups of animals reached similar maximal decreases in SDH activity in striatum and cortex. Remarkably, only the fast decline in SDH activity seen in vulnerable rats was associated with TUNEL labeling. In addition, vulnerable rats developed a region within striatum where SDH activity was fully depleted and a similarly depleted region in CA1 hippocampus. While mice did not develop this region in striatum, some developed one in CA1. These regions of SDH depletion in both structures were associated with widespread TUNEL staining, with maximal labeling at 3 days after 3-NP. The existence of an animal strain resilient to 3-NP suggests that there are mediating factors involved in the preferential vulnerability of striatum to metabolic lesioning. The identification of these factors could provide strategies for therapeutic intervention in HD. PMID- 9743570 TI - NMDA receptors in the inferior colliculus are critically involved in audiogenic seizures in the adult rats with neonatal hypothyroidism. AB - The effects of N-methyl-d-aspartate (NMDA) and non-NMDA receptor antagonists were compared on audiogenic seizures in the rats neonatally exposed to propylthiouracil (PTU). The rats treated with 0.02% PTU through mother's milk during days 0-19 after delivery showed a high incidence of audiogenic seizures consisting of running fit (RF) followed by generalized tonic-clonic seizure (GTCS) after matured. The systemic administration with MK-801, a NMDA receptor antagonist dose-dependently inhibited both RF and GTCS. NBQX (6-nitro-7 sulfamoylbenzo[f]quinoxaline-2,3-dione), a non-NMDA receptor antagonist, when systemically administered, failed to block audiogenic seizures. Audiogenic seizures caused a marked induction of c-fos messenger RNA (mRNA) in septal nucleus, bed nucleus of stria terminalis, amygdaloid nuclei, peripeduncular nucleus, and inferior colliculus, which was almost completely blocked by the pretreatment with MK-801. Bilateral microinjection of MK-801 into the inferior colliculus showed a tendency for inhibiting GTCS, but not RF, whereas CPP (3-(R) 2-carboxypiperazin-4-yl)-propyl-1-phosphonic acid), a competitive NMDA receptor antagonist produced a significant inhibition against both RF and GTCS. These NMDA receptor antagonists administered into cisterna ambience, the floor of which is composed of inferior colliculus and neighboring structures, have shown potent blocking effects on both RF and GTCS. The present results suggest that NMDA receptors in the inferior colliculus, presumably in the subnucleus of external cortex may play the critical role in the initiation of audiogenic seizures in PTU treated rats. PMID- 9743571 TI - Neuroprotection of spinal motoneurons following targeted transduction with an adenoviral vector carrying the gene for glial cell line-derived neurotrophic factor. AB - Application of neurotrophic factors (NFs) to the cut stump of peripheral nerves confers transient (1- to 2-week) neuroprotection of motoneurons from axotomy induced death in neonates. We tested whether lumbar spinal motoneurons would be protected from axotomy-induced death when they were genetically modified to produce NFs in situ. Adenoviral (Adv) vectors carrying neurotrophic factor genes under control of the Rous sarcoma virus long terminal repeat promoter (Adv.RSV nf) or a control vector containing the beta-galactosidase (beta-gal) gene (Adv.RSV-betagal) was injected into the hindlimb muscles of neonatal rats. The Adv were taken up by peripheral nerves and transported to lumbar spinal cord motoneurons where the transgenes were expressed. A fraction (18%) of the motoneurons that projected through the sciatic nerve were transduced with Adv.RSV betagal. Expression of Adv.RSV-betagal was detected in motoneurons after 7 days and 3 weeks, with no evidence of vector- or beta-gal-induced toxicity or inflammation. PCR, immunocytochemistry, and RT-PCR demonstrated transport of the Adv.RSV-nf vectors to motoneurons and their expression. After retrograde transport of an Adv.RSV-nf vector carrying the gene for glial cell line-derived neurotrophic factor, a substantial proportion of the sciatic nerve motoneurons were resistant to axotomy-induced death 7 days and 3 weeks after sciatic nerve transection (56 and 44%, respectively), compared to Adv.RSV-betagal controls (2.5 and 0%, respectively). PMID- 9743572 TI - Pattern and pharmacology of propagating epileptiform activity in mouse cerebral cortex. AB - Multiple extracellular recording electrodes were used to study the intra- and interhemispheric spread of stimulus-evoked epileptiform responses in adult mouse neocortical slices. Bath application of 20 microM bicuculline methiodide induced epileptiform activity that propagated at approximately 0.08 m/s over several millimeters in rostro-caudal and medio-lateral direction within the ipsilateral hemisphere and across the corpus callosum to the contralateral hemisphere. A vertical incision from layer II to subcortical regions did not prevent the spread to remote cortical regions, indicating that layer I plays a major role in the lateral propagation of epileptiform activity. The intra- and interhemispheric spread was not influenced by application of an N-methyl-d-aspartate (NMDA) receptor antagonist, but blocked by an antagonist acting at the (+/-)-alpha-amino 3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA)-type glutamate receptor. The potential role of potassium channel activation in controlling the generation or spread of epileptiform activity was tested by applying the potassium channel opener cromakalim and the serotonin type 1A (5-HT1A) receptor agonist (+/-)-8 hydroxydipropylaminotetralin (8-OH-DPAT) to the disinhibited slices. Whereas cromakalim reduced the neuronal excitability and blocked all epileptiform responses, 8-OH-DAPT did not affect the activity pattern. Our results suggest that propagating epileptiform activity in disinhibited neocortical structures is predominantly mediated by activation of AMPA receptors and controllable by activation of a voltage-dependent potassium current. PMID- 9743573 TI - Amygdalar injection of FeCl3 causes spontaneous recurrent seizures. AB - Rats were microinjected with a 100 mM aqueous solution of ferric chloride into the left amygdaloid body. Behavior was observed and depth electroencephalograms were recorded over the 30 days following injection. All of the FeCl3-injected rats developed isolated epileptiform discharges from the ipsilateral amygdala soon after injection. Within 5 days epileptiform discharges were arising as well from the contralateral amygdala and behavioral seizures were observed. These spontaneous seizures occurred in a pattern associated with stage 4 kindling, with rearing and bilateral forelimb clonus. Seizures persisted during the 30 days of the experiment. Recording from chronically implanted depth electrodes showed development of spike discharges, with recurrent seizures arising from amygdalar regions with propagation into both hippocampi. Aqueous iron is known to initiate lipid peroxidation by free radical mechanisms. Our observations suggest that epileptogenesis followed by chronic, spontaneous seizures could be initiated by deposition of iron-containing compounds into limbic structures of the rat. PMID- 9743574 TI - Effect of glucose deprivation and acute glutamate exposure in cultured retinal cells. AB - The relationship between bioenergetics and the glutamate system was analyzed in a neuronal model of retinal cells in culture, submitted to glucose deprivation and exposed to glutamate for 2 h, and compared with exposure to glutamate in the presence of glucose. Under glucose deprivation, a reduction (about 1.1-fold) in the energy charge of the cells occurred, probably as a result of a decrement (by about 75%) in the cellular redox efficacy, as determined by the 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) test. In the absence of glucose, exposure of retinal cells to 10 microM glutamate potentiated the reduction in the energy charge (by about 1.2-fold) and induced a significant increase in the uptake of 45Ca2+ by the cells (1.3-fold), although no significant changes were observed in the presence of glucose. Under glucose deprivation, 100 microM glutamate caused an irreversible cell membrane damage, as shown by the significant increase in lactate dehydrogenase (LDH) leakage (about 1.8-fold). A significant increase in membrane depolarization, measured by the reduction of [3H]tetraphenylphosphonium+ ([3H]TPP+) uptake, was also observed after glutamate exposure in the absence of glucose. In the presence of glucose, high glutamate concentrations (10 mM) induced a major increase in Ca2+ entry into the cells and membrane depolarization, without affecting the energy charge or cell survival. In contrast, in the absence of glucose, 10 mM glutamate did not alter Ca2+ accumulation by the cells and a smaller decrease in membrane potential occurred, as compared to 100 microM glutamate exposure. Data shown in this study suggest that during a prolonged (2 h) and acute exposure to high glutamate (10 mM), under glucose deprivation conditions, the neuronal systems have "adaptive" mechanisms that allow the survival of cells. These findings may have implications in neuronal degeneration occurring during brain ischemia. PMID- 9743576 TI - Interleukin-10 improves outcome and alters proinflammatory cytokine expression after experimental traumatic brain injury. AB - Traumatic injury to the central nervous system initiates inflammatory processes that are implicated in secondary tissue damage. These processes include the synthesis of proinflammatory cytokines, leukocyte extravasation, vasogenic edema, and blood-brain barrier breakdown. Interleukin-10 (IL-10), a cytokine with antiinflammatory properties, negatively modulates proinflammatory cascades at multiple levels. We examined the hypothesis that IL-10 treatment can improve outcome in a clinically relevant model of traumatic brain injury (TBI). IL-10 was administered via different routes and dosing schedules in a lateral fluid percussion model of TBI in rats. Intravenous administration of IL-10 (100 micrograms) at 30 min before and 1 h after TBI improved neurological recovery and significantly reduced TNF expression in the traumatized cortex at 4 h after injury. Such treatment was associated with lower IL-1 expression in the injured hippocampus, and to a lesser extent, in the injured cortex. Subcutaneous IL-10 administration (100 micrograms) at 10 min, 1, 3, 6, 9, and 12 h after TBI also enhanced neurological recovery. In contrast, intracerebroventricular administration of IL-10 (1 or 6 micrograms) at 15 min, 2, 4, 6, and 8 h after TBI was not beneficial. These results indicate that IL-10 treatment improves outcome after TBI and suggest that this improvement may relate, in part, to reductions in proinflammatory cytokine synthesis. PMID- 9743575 TI - Mitochondria in sporadic amyotrophic lateral sclerosis. AB - Mitochondria are abnormal in persons with amyotrophic lateral sclerosis (ALS) for unknown reasons. We explored whether aberration of mitochondrial DNA (mtDNA) could play a role in this by transferring mitochondrial DNA (mtDNA) from ALS subjects to mtDNA-depleted human neuroblastoma cells. Resulting ALS cytoplasmic hybrids (cybrids) exhibited abnormal electron transport chain functioning, increases in free radical scavenging enzyme activities, perturbed calcium homeostasis, and altered mitochondrial ultrastructure. Recapitulation of defects previously observed in ALS subjects and ALS transgenic mice by expression of ALS mtDNA support a pathophysiologic role for mtDNA mutation in some persons with this disease. PMID- 9743577 TI - High frequency of apolipoprotein E epsilon4 allele in young individuals with very mild Alzheimer's disease-related neurofibrillary changes. AB - The pathological process of initial neurofibrillary (NF) changes underlying Alzheimer's disease (AD) represents the early preclinical phase of the disease. In a small percentage of individuals, these initial NF changes (Braaks' stage I of six stages) may develop at a surprisingly young age. The aim of this study was to determine the impact of apolipoprotein E (ApoE) on the development of such initial NF changes in young individuals. To this end, the ApoE genotypes were determined using a seminested polymerase chain reaction assay followed by restriction isotyping in young individuals (n = 44; mean age of 38 years) with initial NF changes (stage I). The results were compared with ApoE genotypes of age-matched controls (n = 70) devoid of such changes (stage 0). Stage I cases exhibited a significantly higher epsilon4 allele frequency compared to controls (0.18 vs 0.09, P = 0.039). Thus, the present study reveals an association of epsilon4 allele with the early onset of AD-related NF changes in young individuals. This finding underlines the relevance of the asymptomatic phase in the course of AD. PMID- 9743578 TI - Peripheral sensory nerve defects in apolipoprotein E knockout mice. AB - Apolipoprotein E (apoE), a plasma lipoprotein involved in lipid metabolism, is also proposed to have important functions within the central and peripheral nervous systems. To investigate the function of apoE in the peripheral nervous system, we examined the structure of sciatic nerves in apoE-deficient (apoE KO) mice. In the normal peripheral nervous system, apoE is produced by nonmyelinating Schwann cells, suggesting a role for apoE in the support of unmyelinated thermal and nociceptive sensory afferents. Using electron microscopy, we have found that apoE KO mice have abnormal and reduced numbers of unmyelinated axons within the sciatic nerve. ApoE KO unmyelinated axons are irregularly shaped and separated by very little Schwann cell cytoplasm. ApoE KO myelinated fibers and myelin are ultrastructurally normal. Consistent with these morphological findings, apoE KO mice display reduced sensitivity to noxious thermal stimuli. These data provide in vivo support for the hypothesis that apoE promotes neuronal health and survival. PMID- 9743579 TI - Mutation in the alpha-synuclein gene and sporadic Parkinson's disease, Alzheimer's disease, and dementia with lewy bodies. AB - Recently, alpha-synuclein attracted attention when Polymeropoulos and colleagues identified a missense mutation of this gene (Science 276:2045-2047, 1997), which is responsible for a form of early-onset familial Parkinson's disease (PD). Immunohistochemically, alpha-synuclein is localized in Lewy bodies, characteristic brain pathology of PD, dementia with Lewy bodies (DLB), and Alzheimer's disease (AD), suggesting that this protein may link these common neurological diseases. Exploration of the possibility that the same mutation of the alpha-synuclein gene as that in familial PD (Ala53Thr) may also confer susceptibility to sporadic PD, DLB, and AD revealed the mutation in none of the samples of 329 cases and 230 controls examined, suggesting that this mutation is not involved in these neurological diseases. PMID- 9743580 TI - Histochemical localization of glycoconjugates on microglial cells in Alzheimer's disease brain samples by using Abrus precatorius, Maackia amurensis, Momordica charantia, and Sambucus nigra lectins. AB - Four lectins (Abrus precatorius (APA), Maackia amurensis (MAA), Momordica charantia (MCA) and Sambucus nigra (SNA)) have been used to identify glycohistochemically the microglial cells (MGC) activation in autoptic brain samples from Alzheimer's disease (AD) subjects. Three of these lectins (APA, MAA and MCA) have utilized as microglial cell markers for the first time. The identification of new markers for the study of MGC is very important to better understand the role of these type of cells in the metabolic/dismetabolic control of betaA4 in AD which still represents a vexata questio. PMID- 9743581 TI - Insulin/IGF-I rescues immortalized brown adipocytes from apoptosis down regulating Bcl-xS expression, in a PI 3-kinase- and map kinase-dependent manner. AB - Serum deprivation of immortalized brown adipocyte cell line resulted in growth arrest in G0/G1 phases of the cell cycle and apoptosis, as detected either by DNA laddering or by increase in the percentage of hypodiploid cells. Furthermore, apoptosis is concurrent with a dramatic increase in the expression of the proapoptotic protein Bcl-xS, the expression of Bcl-xL remaining almost undetectable. Insulin/insulin-like growth factor (IGF-I) rescued serum-deprived brown adipocytes from apoptosis, decreasing the number of hypodiploid cells and increasing the number of cells undergoing cell cycle progression throughout S and G2/M phases of the cell cycle. Moreover, insulin down-regulated Bcl-xS expression without inducing the expression of Bcl-xL. Both phosphatidylinositol (PI) 3 kinase and mitogen-activated protein kinase (MAPK) pathways are necessary for insulin/IGF-I full survival effect, since the use of specific inhibitors of PI 3 kinase activity (wortmannin or LY294002, at the dose that inhibits PI 3-kinase activity induced by insulin) or MAPK kinase activity inhibitor (PD098059, at the dose that inhibits insulin-induced phosphorylation of MAPK) totally blocked the antiapoptotic effect induced by insulin/IGF-I, respectively. In conclusion, insulin survival effect on immortalized brown adipocytes is associated with inhibition of the Bcl-xS content without changing Bcl-xL, in a PI 3-kinase- and MAP kinase-dependent manner. PMID- 9743582 TI - The accumulation of cyclin-dependent kinase inhibitor p27kip1 is a primary response to staurosporine and independent of G1 cell cycle arrest. AB - The staurosporine-induced G1 cell cycle arrest was analyzed in a variety of cell lines which includes human tumor cell lines and oncogene-transformed NIH3T3 cell lines. All the cell lines which were sensitive to staurosporine-induced G1 arrest contained a functional retinoblastoma protein (pRB). However, when pRB-lacking fibroblast cells derived from pRB knockout mice were tested they were also sensitive to G1 arrest by staurosporine, indicating that the inactivation of pRB alone is not sufficient for the abrogation of staurosporine-induced G1 arrest. In searching for a common event caused by staurosporine, the cyclin-dependent kinase (CDK) inhibitor protein p27kip1 but not p21cip1 was found to accumulate after staurosporine treatment in all the cell lines examined. This accumulation occurred regardless of the induction of the G1 arrest. The result indicates that the accumulation of p27kip1 is the cell's primary response to staurosporine and that the capability of staurosporine to induce G1 arrest depends on the integrity of cell cycle regulatory components which are downstream of p27kip1. PMID- 9743584 TI - MyoD, myogenin, and desmin-nls-lacZ transgene emphasize the distinct patterns of satellite cell activation in growth and regeneration. AB - Although satellite cell differentiation is involved in postnatal myogenesis from growth to posttrauma regeneration, the early stages of this process remain unclear. This study investigated pHuDes-nls-lacZ transgene activity, as revealed by X-gal staining and the accumulation of MyoD, myogenin, endogenous desmin, and myosin, in order to determine whether satellite cells share the same activation program during growth and regeneration. After birth, skeletal myonuclei in which myogenin expression was limited were briefly characterized by transgene activity. Satellite cells were only evidenced by MyoD and slow myosin accumulation, but failed to initiate transgene expression. After freeze trauma, satellite cell activation led to MyoD, myogenin, and desmin expression. Subsequently, when myosin expression occurred, transgene activation was apparent in regenerating structures, with more intense X-gal staining in mononucleated cells than regenerating myotubes. After the second week posttrauma, only desmin and myogenin expression were maintained in regenerating structures. In culture, the behavior of satellite cells showed that desmin expression was committed before transgene activation occurred, i.e., concurrently with MyoD, myogenin, myosin expression, and the first fusion events. Quantitative analysis confirmed the discrepancy between endogenous desmin and transgene expression and demonstrated the close correlation between transgene activation and the fusion index. Our results strongly suggest that satellite cells promote distinct pathways of myogenic response during growth and regeneration. PMID- 9743583 TI - Nuclear distribution of human DNA topoisomerase IIbeta: a nuclear targeting signal resides in the 116-residue C-terminal tail. AB - We have analyzed the subcellular distribution of the beta isoform of human topoisomerase II using both isoform-specific antisera and an epitope-tagging approach. Previous immunocytochemical studies have yielded differing results with one reporting this isoform to be predominantly nucleolar. Later studies seem to refute this finding, as do our results with isoform-specific antisera reported here. Epitope tagging minimizes potential complications arising from the use of anti-topoisomerase II antisera that may recognize epitopes that are modified or masked in vivo and could lead to misleading results in immunocytochemical studies. A second strength of this approach is that it allowed a comparison with similarly tagged control proteins (derived from the nucleolar transcription factor UBF) that were known to localize unambiguously to the cytoplasmic, nucleoplasmic, or nucleolar compartments. We report that the C-terminal domain of topoisomerase IIbeta fused to a beta-galactosidase tag localizes to the nucleus (but not the nucleolar compartment) and that this is indistinguishable from the localization of native topoisomerase IIbeta detected by isoform-specific antisera. Further analysis revealed that the nuclear localization determinant lies within the 116-residue C-terminal tail of human topoisomerase IIbeta. PMID- 9743585 TI - Actin polymerization is required for negative feedback regulation of epidermal growth factor-induced signal transduction. AB - Epidermal growth factor (EGF) induces rapid actin filament assembly in the membrane skeleton of a variety of cells. To investigate the significance of this process for signal transduction, actin polymerization is inhibited by dihydrocytochalasin B (CB). CB almost completely abolishes EGF-induced actin polymerization, as assessed by quantitative confocal laser scanning microscopy. Under these conditions, EGF induces enhanced EGF receptor (EGFR) tyrosine kinase activity, as well as superinduction of the c-fos proto-oncogene. These data suggest that EGF-induced actin polymerization may be important for negative feedback regulation of signal transduction by the EGFR. The phosphorylation of Thr654 by protein kinase C (PKC) is a well-characterized negative feedback control mechanism for signal transduction by the EGFR tyrosine kinase. A synthetic peptide, corresponding to the regions flanking Thr654 of the EGFR, is used to analyze EGF stimulated PKC activity by incorporation of 32P into the peptide. Cotreatment of cells with CB and EGF results in a complete loss of EGF induced phosphorylation of the peptide. These data suggest that actin polymerization is obligatory for negative feedback regulation of the EGFR tyrosine kinase through the C-kinase pathway. PMID- 9743586 TI - Hepatocyte growth factor enables enhanced integrin-cytoskeleton linkage by affecting integrin expression in subconfluent epithelial cells. AB - Hepatocyte growth factor (HGF) exerts mitogenic and motogenic effects in different cell types. In the epithelial cell line mHepR1 we found that HGF induced pronounced alterations in cell morphology and promoted cell adhesion and spreading. To analyze the mechanisms how HGF affects these integrin mediated functions we studied the physical linkage of integrins with the cytoskeleton. First we found that HGF increased the expression of different integrin subunits in subconfluent cells and influenced the distribution of integrins on the cell surface. To address the physical association of integrins with the cytoskeleton we analyzed Triton X-100-extracted cell fractions using flow cytometry. Here we show that cultivation of the cells with HGF for 24 h prior to integrin cross linking significantly enhanced the cytoskeletal anchorage of integrins. To further find out whether HGF directly induces an integrin-cytoskeleton link without subsequent cross-linking we added HGF to suspended cells but failed to detect cytoskeletally immobilized integrins in the detergent-insoluble cell fraction which could be related to the absence of a calcium response induced by HGF. Overall, the results indicate that HGF promotes the physical linkage of integrins to the cytoskeleton which requires additional stimulation of integrins. PMID- 9743587 TI - H-ras induces glucose uptake in brown adipocytes in an insulin- and phosphatidylinositol 3-kinase-independent manner. AB - Fetal brown adipocytes (parental cells) expressed mainly Glut4 mRNA glucose transporter, the expression of Glut1 mRNA being much lower. At physiological doses, insulin stimulation for 15 min increased 3-fold glucose uptake and doubled the amount of Glut4 protein located at the plasma membrane. Moreover, phosphatidylinositol (PI) 3-kinase activity was induced by the presence of insulin in those cells, glucose uptake being precluded by PI 3-kinase inhibitors such as wortmannin or LY294002. H-raslys12-transformed brown adipocytes showed a 10-fold higher expression of Glut1 mRNA and protein than parental cells, Glut4 gene expression being completely down-regulated. Glucose uptake increased by 10 fold in transformed cells compared to parental cells; this uptake was unaltered in the presence of insulin and/or wortmannin. Transient transfection of parental cells with a dominant form of active Ras increased basal glucose uptake by 5 fold, no further effects being observed in the presence of insulin. However, PI 3 kinase activity (immunoprecipitated with anti-alphap85 subunit of PI 3-kinase) remained unaltered in H-ras permanent and transient transfectants. Our results indicate that activated Ras induces brown adipocyte glucose transport in an insulin-independent manner, this induction not involving PI 3-kinase activation. PMID- 9743588 TI - Cell fusion studies to examine the mechanism for etoposide resistance in Chinese hamster V79 spheroids. AB - When exposed to etoposide, the outer cells from Chinese hamster V79 spheroids are about 10 times more resistant to DNA strand breaks and cell killing than V79 cells grown as monolayers. Previous results have shown that the outer cells of both spheroids and monolayers grow at the same rate and contain the same amount and activity of the target enzyme, topoisomerase II. In order to examine possible mechanisms for this resistance, cell fusion studies were conducted with fluorescent dye-tagged monolayer and spheroid cells. Fused cells were exposed for 30 min to 1.2 microg/ml etoposide and then separated using fluorescence-activated cell sorting into binucleate cells consisting of two monolayer cells, two spheroid cells, or a mixed doublet consisting of one cell of each type. Individual sorted cell doublets were examined for the presence of etoposide induced DNA strand breaks using the alkaline comet assay. As expected, doublets of monolayer cells were sensitive to etoposide and doublets of spheroid cells were resistant. However, mixed doublets were as resistant to DNA damage by etoposide as spheroid doublets. In comparison, when etoposide- or adriamycin resistant V79 monolayer cells were fused to the parent monolayer cells, the expected intermediate sensitivity to etoposide was observed for the mixed doublets. We conclude that etoposide resistance associated with the outer cells of spheroids can be "transferred" to produce resistance in monolayer cells. Rapid changes in phosphorylation that can affect topoisomerase II activity or localization, or that can alter chromatin structure, are suggested as possible mechanisms of resistance. In support of this hypothesis, topo IIalpha phosphorylation was at least 10 times greater in monolayers than in the outer cell layer of spheroids. PMID- 9743589 TI - Dynamic interactions between splicing snRNPs, coiled bodies and nucleoli revealed using snRNP protein fusions to the green fluorescent protein. AB - The U1, U2, U4/U6, and U5 small nuclear ribonucleoproteins (snRNPs) are subunits of splicing complexes that remove introns from mRNA precursors. snRNPs show a complex, transcription-dependent localization pattern in the nucleoplasm of mammalian cells that results from their association with several distinct subnuclear structures, including interchromatin granule clusters, perichromatin fibrils, and coiled bodies. Here we report the analysis of snRNP localization and interaction with the coiled body in live human cells using fusions of snRNP proteins and p80 coilin to the Green Fluorescent Protein (GFP). Despite the large size of the GFP tag, GFP fusions to both the core snRNP SmE and U1 specific U1A proteins assemble into snRNP particles and give an identical nuclear localization pattern to their endogenous counterparts. GFP-coilin localizes specifically to coiled bodies in a transcription-dependent fashion and provides an accurate marker for coiled bodies in a variety of human cell lines. Treatment of cells with the selective ser/thr-protein phosphatase inhibitor, okadaic acid, causes both GFP-snRNP and GFP-coilin proteins to accumulate within nucleoli, but does not result in nucleolar accumulation of the GFP-fused non-snRNP protein splicing factor ASF/SF2. In all four human cell lines tested, expression of a GFP-fused p80 coilin mutant with a single serine to aspartate substitution also caused nucleolar accumulation of splicing snRNPs and coilin, but not ASF/SF2, in structures resembling coiled bodies when viewed by electron microscopy. This work establishes an experimental system for analyzing snRNP trafficking in living cells and provides evidence that a reversible protein phosphorylation mechanism is involved in regulating interaction of snRNPs and coiled bodies with the nucleolus. PMID- 9743590 TI - Immunopurification of a sarcomeric junctional protein complex containing GAPDH. AB - We have isolated a monoclonal antibody, P4B2, which localizes to multiple anchorage junctions, namely, a subset of focal adhesions, the Z-disk of muscle, and neuromuscular junctions. Immunopurification of the antigen to this antibody from chicken brain tissue yielded a complex of three prominent proteins with mobilities of 36, 30, and 18 kDa. Amino acid sequencing of the purified proteins identified the 36-kDa protein as glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The other two protein bands were heterogeneous, containing proteins found in the synaptic vesicle fusion core complex. Immunolocalization of P4B2 antigen in developing cultured muscle cells showed that the antigen is incorporated into Z-lines soon after the sarcomeric architecture was positive for alpha-actinin. Together, the data indicate the P4B2 antigen is part of a unique GAPDH-containing protein complex that may be involved in reinforcement of established cytoskeletal structures. PMID- 9743591 TI - Concomitant changes in polyamine pools and DNA methylation during growth inhibition of human colonic cancer cells. AB - The effects of CGP 48664 and DFMO, selective inhibitors of the key enzymes of polyamine biosynthesis, namely, of S-adenosylmethionine decarboxylase (AdoMetDC) and ornithine decarboxylase (ODC), were investigated on growth, polyamine metabolism, and DNA methylation in the Caco-2 cell line. Both inhibitors caused growth inhibition and affected similarly the initial expression of the differentiation marker sucrase. In the presence of the AdoMetDC inhibitor, ODC activity and the intracellular pool of putrescine were enhanced, whereas the spermidine and spermine pools were decreased. In the presence of the ODC inhibitor, the AdoMetDC activity was enhanced and the intracellular pools of putrescine and spermidine were decreased. With both compounds, the degree of global DNA methylation was increased. Spermine and spermidine (but not putrescine) selectively inhibited cytosine-DNA methyltransferase activity. Our observations suggest that spermidine (and to a lesser extent spermine) controls DNA methylation and may represent a crucial step in the regulation of Caco-2 cell growth and differentiation. PMID- 9743592 TI - Activation of the insulin-like growth factor type 1 receptor by deletion of amino acids 870-905. AB - We have created a deletion mutant of the insulin-like growth factor type 1 receptor (IGF-1 R) which lacks the 36 amino acids (aa) immediately N-terminal to the transmembrane domain (Delta870-905 IGF-1 R). This region has been reported to have a negative effect on the transforming potential of an avian sarcoma virus gag-IGF-1 R fusion protein. We have sought to determine whether this region plays a similar role in the intact IGF-1 R. Analysis of the tyrosine kinase activity of the Delta870-905 IGF-1 R shows that the mutant receptor is autophosphorylated without IGF-1 stimulation, indicating that the tyrosine kinase domain is constitutively active. In addition, processing of the receptor is decreased, resulting in accumulation of a high molecular weight proreceptor containing both alpha and beta-subunits. A well-characterized substrate of the IGF-1 R, IRS-1, is constitutively phosphorylated by the Delta870-905 IGF-1 R and phosphoinositide (PI) 3-kinase activity, which is normally activated by the phosphorylation of IRS 1 following IGF-1 stimulation, is increased even in the absence of IGF-1. A second intracellular signal pathway normally activated by IGF-1, the MAP kinase pathway, showed no increase in activity in the absence of IGF-1. The Delta870-905 IGF-1 R promoted cell proliferation only in the presence of IGF-1. We conclude that this deletion increases the basal activity of the IGF-1 receptor tyrosine kinase and activates PI 3-kinase, but is unable to stimulate MAP kinase in the absence of ligand. These results confirm those seen in the gag-IGF-1 R fusion protein and indicate that aa 870-905 exert a negative effect on the tyrosine kinase domain of the beta-subunit of the IGF-1 R. PMID- 9743593 TI - Cloning of rat vitamin K-dependent gamma-glutamyl carboxylase and developmentally regulated gene expression in postimplantation embryos. AB - Vitamin K-dependent carboxylase catalyzes the posttranslational modification of glutamate to gamma-carboxyglutamate (Gla) in its substrates, the vitamin K dependent proteins (VKDPs). This modification is required for the activities of the VKDPs. Recent evidence demonstrates previously unrecognized roles for VKDPs as signaling molecules important in the regulation of cell growth, adhesion, and apoptosis, suggesting developmental functions for VKDPs and hence the carboxylase. The tissue distribution and functions of carboxylase in development are unknown. In this study, we isolated and characterized the full-length cDNA encoding the rat carboxylase and analyzed, at the cellular level, the expression of this gene in rat embryos by in situ hybridization. We demonstrate that the expression of this gene is highly regulated in a developmental and tissue specific manner. Hepatocytes, the major site of synthesis of VKDPs of blood coagulation, express carboxylase mRNA late in gestation, in contrast to the central nervous system, mesenchymal, and skeletal tissues which express carboxylase mRNA early during rat embryogenesis. The tissue-specific temporal expression of the carboxylase gene during embryogenesis indicates that vitamin K dependent carboxylation and the formation of Gla is developmentally regulated. These studies suggest that vitamin K-dependent carboxylation is an important modulator of embryonic VKDP function. PMID- 9743595 TI - Characterization of the A549 cell line as a type II pulmonary epithelial cell model for drug metabolism. AB - Multiple cell types contribute to the pulmonary barrier including Type I and Type II alveolar epithelium. The objective of this research was to establish and characterize an in vitro model of Type II alveolar epithelium using the A549 human lung adenocarcinoma cell line. A549 cells form confluent monolayers with Type II characteristic morphology and tannic acid staining for typical lamellar bodies. A549 cells possess P450 IA1 and P450 IIB6 as determined by Western blots. Both CYPIA1 and CYPIIB6 P450 isozymes were determined to be functional with the fluorescent resorufin assay. Only the IA1 isozyme was observed to be inducible with selected polycyclic hydrocarbons. Uptake and transport experiments were carried out in cluster plates and in Snapwells. Cationized ferritin, a nonspecific absorbtive marker, was found to be taken up by the cells in a concentration-, time-, and temperature-dependent fashion. Lucifer yellow, a fluid phase marker, was not internalized by the A549 cells. Transferrin, a representative receptor-mediated endocytic marker, was found to be taken up by the cells in a concentration-dependent and competitive fashion. Transport experiments involving fluorescein-transferrin also showed that A549 monolayers were polarized, with a greater amount of intracellular transferrin being transported out of the basolateral side of the cells. The experimental data agree favorably with literature for primary cultures of Type II pulmonary epithelial cells. These results indicated that the A549 cell line may be useful for the studying the metabolic and macromolecule processing contributions of alveolar Type II cells to mechanisms of drug delivery at the pulmonary epithelium. PMID- 9743594 TI - EGF modulates expression of STAT5 in mammary epithelial cells. AB - HC11 mouse mammary epithelial cells are capable of differentiating in vitro. By growing cells in EGF-containing medium, and upon confluence withdrawing EGF, these cells become competent at responding to lactogenic hormone treatment and expressing milk proteins. We found that during proliferation and at confluence STAT5A and STAT5B proteins were expressed at equal levels or with STAT5B being predominant. In competent cells, expression levels of STAT5A and STAT5B increased markedly with STAT5A now being the predominant form, an expression pattern resembling the expression patterns of STAT5 proteins seen during mammary gland differentiation in vivo. This suggests that EGF has a suppressive effect on STAT5 expression, in particular, STAT5A, which we conclude to be mediated through ras/raf/MEK/MAPK pathway and to a lesser extent through a PI3-kinase-mediated pathway. Furthermore, we also found that EGF regulated a nuclear phosphatase capable of dephosphorylating tyrosine-phosphorylated STAT5. Our data show that HC11 cells have retained the expression patterns of STAT5 proteins seen in vivo. This makes HC11 cells useful for studying molecular mechanisms regulating expression of STAT factors and their participation in differentiation processes of mammary gland. PMID- 9743596 TI - Antiproliferative effect of the tripeptide pyroGlu-Phe-GlyNH2 on murine melanocytes: transitory delay of cell growth in vitro and the cell cycle specificity. AB - Cell growth and differentiation in melanocyte cell populations are regulated by a wide range of bioactive substances. Recently, the tripeptide pyroGlu-Phe-GlyNH2 which inhibits melanocyte growth in vitro was identified in both murine nontransformed melanocytes and malignant melanoma cells. The present study was undertaken to investigate the cell cycle specificity as well as the growth inhibitory profile of the tripeptide after a single or repeated administration to melanocyte cultures. Dose-related effects of the peptide were studied using three different bioassay systems: estimation of cell number, DNA synthesis, and cell flux into mitosis. Growth of melanocyte cultures as well as melanocyte mitotic activity were found to be reduced significantly by the tripeptide at two separate dose levels (10(-11) and 10(-14)-10(-15) M). Growth inhibition of melanocyte population did not last long: less than 36 h after the first and less than 24 h after the second peptide addition to the cultures. The level of DNA synthesis in melanocytes remained unchanged after a single peptide administration. The findings indicate that the tripeptide pyroGlu-Phe-GlyNH2 causes transitory delay of cell growth in cultured melanocyte population resulting from a reversible inhibition of melanocyte transition from the G2-phase of the cell cycle into mitosis. PMID- 9743597 TI - Laminin-1 and the RKRLQVQLSIRT laminin-1 alpha1 globular domain peptide stimulate matrix metalloproteinase secretion by PC12 cells. AB - Here we have investigated the ability of laminin-1 and specific laminin-1-derived synthetic peptides to stimulate neuronal cell matrix metalloproteinase secretion. Zymographic analysis of conditioned media from laminin-1-treated PC12 and NG108 15 cells revealed a 72-kDa matrix metalloproteinase which was not secreted by untreated cells. Laminin-1 alpha1 chain-derived synthetic peptides, AASIKVAVSADR (LAM-L) and RKRLQVQLSIRT (AG-73), also stimulated PC12 cell secretion of a 72-kDa matrix metalloproteinase. We further investigated the structural requirements of AG-73 for cell attachment, neurite outgrowth, and matrix metalloproteinase secretion using a series of AG-73 analogs that had single amino acids substituted with alanine. At the substrate levels tested, the AG-73 peptide promoted the adhesion of 67% of the PC12 cells and neurite outgrowth in 71% of the PC12 cells. Substitutions in any one of the amino acids within the central LQVQ sequence resulted in a large reduction in cell attachment whereas substitution in the carboxyl terminal proximal amino acids L, S, and R had little effect on attachment. Alanine substitution of any of the amino terminal proximal LQV amino acids and the carboxyl terminal L, I, and R residues resulted in a 65-91% reduction in neurite outgrowth. These data demonstrate that the sequence requirements for cell attachment and neurite outgrowth were not necessarily coupled but that the sequence requirements for neurite outgrowth and matrix metalloproteinase secretion were identical. We conclude that laminin-1 is able to stimulate neuronal cells to secrete a matrix metalloproteinase. Further, this study identifies the LQVXLXIR laminin-1 alpha1 globular domain peptide to be capable of stimulating both neurite outgrowth and matrix metalloproteinase secretion. PMID- 9743598 TI - Induction of apoptosis in cardiac myocytes by an A3 adenosine receptor agonist. AB - The effects of the selective adenosine (ADO) A3 receptor agonist IB-MECA (N6-(3 iodobenzyl)adenosine-5'-N-methylcarboxamide) on cultured newborn rat cardiomyocytes were examined in comparison with ADO, the ADO A1 receptor selective agonist R-PIA (N6-R-phenylisopropyladenosine), or the ADO A3 selective antagonist MRS 1191 (3-ethyl-5-benzyl-2-methyl-6-phenyl-4-phenylethynyl-1, 4-(+/ )-dihydropyridine-3,5 dicarboxylate), using digital image analysis of Feulgen stained nuclei. At high concentration, IB-MECA (>/=10 microM ) and ADO (200 microM) induced apoptosis; however, R-PIA or MRS 1191 did not have any detectable effects on cardiac cells. In addition, DNA breaks in cardiomyocytes undergoing apoptosis following treatment by IB-MECA were identified in situ using the nick end labeling of DNA ("TUNEL"-like) assay. In the presence of >/=10 microM IB MECA, disorder in the contraction waves appeared, and a decrease in the frequency of beats was observed. Analysis with light microscopy revealed that the number of contracting cells decreased in a concentration-dependent manner. The A3 receptor agonist IB-MECA caused an increase in intracellular free calcium concentration ([Ca2+]i). The drug produced a rapid rise followed by a sustained increase in [Ca2+]i, which lasted for 40-60 s. Finally, cessation of beating and Ca2+ transients were observed. Full recovery of contractile activity and rhythmical Ca2+ transients were observed 15-20 min after IB-MECA treatment. The induction of apoptosis in the cardiocytes by IB-MECA led to the appearance of features of apoptotic nuclei: the onset of condensation, compacting, and margination of nuclear chromatin. These effects were accompanied by the disintegration of the structural framework of the nucleus and nuclear breakdown. The results suggest that activation of the A3 adenosine receptor may participate in the process of apoptosis in cardiomyocytes. PMID- 9743599 TI - Spatial distributions of early and late replicating chromatin in interphase chromosome territories. AB - The surface area of chromosome territories has been suggested as a preferred site for genes, specific RNAs, and accumulations of splicing factors. Here, we investigated the localization of sites of replication within individual chromosome territories. In vivo replication labeling with thymidine analogues IdUrd and CldUrd was combined with chromosome painting by fluorescent in situ hybridization on three-dimensionally preserved human fibroblast nuclei. Spatial distributions of replication labels over the chromosome territory, as well as the territory volume and shape, were determined by 3D image analysis. During late S phase a previously observed shape difference between the active and inactive X chromosome in female cells was maintained, while the volumes of the two territories did not differ significantly. Domains containing early or mid to late replicating chromatin were distributed throughout territories of chromome 8 and the active X. In the inactive X-chromosome early replicating chromatin was observed preferentially near the territory surface. Most important, we established that the process of replication takes place in foci throughout the entire chromosome territory volume, in early as well as in late S-phase. This demonstrates that activity of macromolecular enzyme complexes takes place throughout chromosome territories and is not confined to the territory surface as suggested previously. PMID- 9743600 TI - Permeable FGF-1 nuclear localization signal peptide stimulates DNA synthesis in various cell types but is cell-density sensitive and unable to support cell proliferation. AB - An earlier report indicated that a 26-amino-acid peptide (SA), comprised of the nuclear localization signal (NLS) of fibroblast growth factor-1 (FGF-1) and a membrane-permeable peptide, was able to stimulate DNA synthesis after it was taken up by NIH3T3 fibroblasts. Here, we report that SA, but not a mutant with the NLS motif destroyed, induced DNA synthesis in BALB/c3T3 murine fibroblasts, human vascular endothelial (HUVE) cells, and primary cultured hepatocytes, although the activity was weaker than that of FGF-1. The kinetics of SA-induced DNA synthesis and G1 cyclin expression were similar to those elicited by FGF-1, indicating that SA induces cell cycle progression. Kinetic analysis also suggested that SA stimulates only a fraction of the DNA replication in BALB/c3T3 cells. At high cell densities, SA-induced G1 cyclin expression and DNA synthesis were more strongly inhibited than those induced by FGF-1. SA did not induce cell division in HUVE and BALB/c3T3 cells and did not interfere with FGF-1-stimulated proliferation of HUVE cells. These results indicate that SA is able to partially induce cell cycle progression through a contact-inhibition sensitive signaling pathway, but it is insufficient to support cell mitosis. We also suggest that signaling by SA does not interfere with that of FGF-1. PMID- 9743601 TI - Modulation of cell-cell adherens junctions by surface clustering of the N cadherin cytoplasmic tail. AB - Cadherins mediate the formation of cell-cell adherens junctions (AJ) by homophilic interactions through their extracellular domains as well as by interacting with the actin cytoskeleton via their cytoplasmic portions. Cadherin clustering initiates cytoplasmic signaling that results in the assembly of structural components into cell-cell AJ. To elucidate the function of the cytoplasmic tail of cadherins in initiating the assembly signal, we generated and characterized a chimeric cadherin tail fused to an inert transmembrane anchor. The chimera enabled us to cluster the cadherin cytoplasmic tail in the absence of extracellular portions of the molecule. The transfected cadherin tail chimera localized to cell-cell AJ of epithelial cells, indicating that the submembrane junctional plaque has the capacity to recruit additional cadherins, with no involvement of their extracellular domains. Expression of the chimera in cells of mesenchymal origin resulted in dominant negative effects on the formation of cell cell AJ. Surface clustering of cadherin cytoplasmic tails induced the recruitment of components and structural assembly of cell-cell AJ, thereby reversing the initial dominant-negative effects. We conclude that the cadherin cytoplasmic tail contains information required to direct the molecule to cell-cell AJ. Its function as modulator of cell-cell AJ depends on cell type and on whether the tail is clustered. PMID- 9743602 TI - Lipopolysaccharide-induced NF-kappaB activation in human endothelial cells involves degradation of IkappaBalpha but not IkappaBbeta. AB - We studied the signal transduction pathways involved in NF-kappaB activation and the induction of the cytoprotective A20 gene by lipopolysaccharide (LPS) in human umbilical vein endothelial cells (HUVEC). LPS induced human A20 mRNA expression with a maximum level 2 h after stimulation. The proteasome inhibitor N-acetyl leucinyl-leucinyl-norleucinal-H (ALLN) and the tyrosine kinase inhibitor herbimycin A (HMA) blocked A20 mRNA expression and partially inhibited NF-kappaB DNA-binding activity induced by LPS treatment. LPS induced IkappaBalpha degradation at 30-60 min after treatment, but did not induce IkappaBbeta degradation up to 120 min. In contrast, TNF-alpha rapidly induced IkappaBalpha degradation within 5 min and IkappaBbeta degradation within 15 min. Cycloheximide did not prevent LPS-induced IkappaBalpha degradation, indicating that newly synthesized proteins induced by LPS were not involved in LPS-stimulated IkappaBalpha degradation. LPS-induced IkappaBalpha degradation was inhibited by ALLN, confirming that ALLN inhibits NF-kappaB activation by preventing IkappaBalpha degradation. Of note, HMA also inhibited LPS-induced IkappaBalpha degradation. However, tyrosine phosphorylation of IkappaBalpha itself was not elicited by LPS stimulation, suggesting that tyrosine phosphorylation of a protein(s) upstream of IkappaBalpha is required for subsequent degradation. We conclude that in HUVEC, LPS induces NF-kappaB-dependent genes through degradation of IkappaBalpha, not IkappaBbeta, and propose that this degradation is induced in part by HMA-sensitive kinase(s) upstream of IkappaBalpha. PMID- 9743603 TI - Induction of a regular nuclear lattice by overexpression of NuMA. AB - Transient overexpression of nuclear mitotic apparatus protein (NuMA) in HeLa cells results in ordered lattices which can fill the nucleus and which are stable to detergent extraction. Electron microscopy reveals a quasi-hexagonal organization with an average spacing between the vertices of approximately 170 nm and short 6-nm-diameter rods connecting the vertices. Overexpression of a NuMA construct with an in-frame addition in the coiled-coil domain shows hexagons with the spacing increased by 42% while constructs with deletions in the coiled-coil domain yield hexagons with the spacing decreased by 40 and 19%. NuMA constructs truncated at residue 2005 or 2030 in the tail domain cause a drastic reorganization of nuclear components with relocation of the DNA, histone H1, and nucleoli to the nuclear rim. A construct lacking the head and much of the coiled coil region also affects nuclear organization. In contrast, NuMA constructs truncated at residue 1950 or 1935 which lack the nuclear localization signal display normal nuclear structure but form cytoplasmic aggregates which also display hexagonal organization. Immunoelectron microscopy confirms that the nuclear lattices are built from NuMA. We discuss the importance of the different domains of NuMA for building the ordered in vivo lattices and whether NuMA could play a structural role in the architecture of the normal interphase nucleus. PMID- 9743604 TI - ICAD/DFF regulator of apoptotic nuclease is nuclear. AB - In nonapoptotic cells, the caspase-activated DNase CAD/CPAN is associated with a regulatory subunit, ICAD/DFF, that binds to it and blocks its enzymatic activity. It has been proposed that a major function of ICAD is to restrain CAD in the cytoplasm in healthy cells. The experiments presented here demonstrate that rather than being cytoplasmic, a GFP-ICAD fusion protein is nuclear in healthy human, pig, and chicken cells. Furthermore, immunoblots using antibodies to murine ICAD confirm the presence of endogenous ICAD and the marker protein DNA topoisomerase I in human nuclei, while tubulin was found solely in the cytosolic fraction. Since ICAD is located in cell nuclei, it is unlikely that the protein functions primarily in the cytoplasm either as an anchor for CAD/CPAN or as a factor that enters the nucleus following caspase cleavage in order to activate resident endonucleases. PMID- 9743605 TI - Introducing rapid communications in experimental cell research PMID- 9743606 TI - New methods for detection of anti-nuclear antibodies. AB - Many different autoantibodies which react with a variety of different nuclear and cytoplasmic antigens have been described. Detection of some these antibodies has been shown to be clinically useful in a number of different autoimmune diseases. For many years, the detection of most of the clinically relevant antibodies was done with by immunofluorescence on tissue substrates and human cultured cell lines. Within the past few years, a number of technical advances has now made it possible to convert to enzyme immunoassay. The paper reviews the clinically relevant antibodies and discusses the variety of new methods which are now available for ANA detection in diagnostic laboratories. PMID- 9743607 TI - beta-Defensins: endogenous antibiotics of the innate host defense response. PMID- 9743609 TI - Human neutrophils express the interleukin-15 receptor alpha chain (IL-15Ralpha) but not the IL-9Ralpha component. AB - The interleukin-15 receptor (IL-15R) is composed of at least three chains, namely gammac, IL-2Rbeta, and the recently identified IL-15Ralpha, while the IL-9R complex consists of gammac and a subunit designated IL-9Ralpha. Our previous work and that of others have shown that human neutrophils express gammac and IL-2Rbeta (two components shared with IL-2R) but not IL-2Ralpha and that IL-15 is a neutrophil agonist, whereas IL-2 is not. In this study, using flow cytometry with a specific anti-human IL-15Ralpha, we show for the first time that human neutrophils express surface IL-15Ralpha. Although we previously found that IL-15 is a neutrophil agonist, our present work shows that IL-15 does not trigger superoxide production nor cell spreading onto glass. In addition, we report that human neutrophils do not respond to IL-9 with respect to the functions/responses studied, namely, superoxide production, spreading onto glass, cell shape changes, phagocytosis, RNA synthesis, and apoptosis. Further, our results show that neutrophils do not express IL-9Ralpha as assessed by flow cytometry with a specific anti-human IL-9Ralpha antibody that stains the transfected cell line BW h9R used as positive control. Finally, our results indicate that gammac expression was not modulated and remained stable for up to 24 h when neutrophils were stimulated with all currently known "gammac users," namely, IL-2, IL-4, IL 7, IL-9, and IL-15. We conclude that human neutrophils express all IL-15R components on their surface, including IL-15Ralpha, that IL-15 activates human neutrophils (as the IL-4 neutrophil agonist) by a mechanism which does not involve upregulation of gammac cell surface expression, and that IL-9 is not a neutrophil agonist as demonstrated by the inability to modulate the tested functions/responses that correlate with lack of the IL-9R component, namely, IL 9Ralpha. PMID- 9743608 TI - The possible role of interleukin (IL)-12 and interferon-gamma-inducing factor/IL 18 in protection against experimental Mycobacterium leprae infection in mice. AB - Cell-mediated immunity participates in host defense against mycobacterial infection. Both interleukin 12 (IL-12) and interferon-gamma-inducing factor (IGIF/IL-18), produced mainly by macrophages, play a critical role in expression of cell-mediated immunity. To investigate the role of IL-12 and IGIF/IL-18 in vivo, we examined cytokine profile, bacterial growth, and the potential benefit of cytokine therapy in susceptible and resistant mice infected with Mycobacterium leprae. The early expression of IL-12 p40 and IGIF/IL-18 at the site of inoculation was found in resistant mice 3-72 h after the infection, but not in susceptible mice. Both strains of mice did not show expression of IFN-gamma and IL-4. IL-12 administration resulted in a significant reduction of bacterial counts in mice with established M. leprae infection. The results imply that susceptible mice exhibit decreased expression of type 1 helper T (Th1) response without reciprocal increased Th2 response and show responsiveness to exogenous IL 12. IL-12 therapy may be a possible rationale for treatment of M. leprae infection. PMID- 9743610 TI - Recovery from Guillain-Barre syndrome is associated with increased levels of neutralizing autoantibodies to interferon-gamma. AB - Guillain-Barre syndrome (GBS) is an immune-mediated demyelinating disease of peripheral nerves that is often preceded by an infection and is usually self restricted. The Th1 cytokine interferon-gamma (IFN-gamma) is thought to be disease-promoting in organ-specific autoimmune diseases. We report the spontaneous induction of IFN-gamma and a mechanism involving the generation of neutralizing autoantibodies (Aabs) to IFN-gamma that may regulate the disease. Numbers of cells spontaneously secreting IFN-gamma in peripheral blood were augmented in GBS, in particular at the peak of clinical disease, and decreased during recovery. This decrease was associated with elevated serum concentrations of IgG Aabs to IFN-gamma. These Aabs specifically bound to IFN-gamma and neutralized its effects in a biological assay. Aabs to IFN-gamma are proposed to be another important regulatory mechanism in IFN-gamma-driven GBS. PMID- 9743611 TI - Immaturity of lymphocytes in the metastatic lesions of thymoma. AB - Thymoma is a thymic epithelial tumor which often contains a large number of immature T cells. Although the metastatic lesions are also associated with abundant lymphocytes, their characteristics have not been assessed in detail. In this study, the phenotype was analyzed and compared with those in their primary lesions. Nine metastatic thymomas were obtained from seven patients. In the metastatic lesions, CD1a+ cells and CD4(+)CD8(+) cells accounted for 77.7 +/- 10.6 and 52.3 +/- 15.8% of all the lymphocytes, respectively. In five primary lesions and their metastatic lesions, CD3(-)CD4(+)CD8(-) cells accounted for 23.9 +/- 16.9 and 45.2 +/- 15. 5% of the CD4(+)CD8(-) cells, respectively. CD69 was expressed on 70. 9 +/- 9.5 and 53.1 +/- 11.8% of the CD4(+)CD8(-) cells, respectively. These results indicate that the metastatic lesions of thymoma are associated with abundant immature T cells which are phenotypically less mature than those in their primary lesions. PMID- 9743612 TI - Decline in total T cell count is associated with onset of AIDS, independent of CD4(+) lymphocyte count: implications for AIDS pathogenesis. AB - We previously reported that blind T cell homeostasis, in which the total T cell count is maintained but the CD4(+) and CD8(+) subset composition of the T cells can vary, fails approximately 1.5 to 2.5 years before the onset of AIDS. The present study was premised on the hypothesis that if failure of T cell homeostasis (i.e., a decline in total T cell counts) is important in the pathogenesis of AIDS, it should be a significant predictor of AIDS after controlling for the CD4(+) lymphocyte count. Data from 1556 homosexual men with sufficient sequential T cell subset measurements were evaluated, representing 11,988 person-visits in men with known clinical outcomes over a period of more than 10 years. Using regression models that incorporated CD4(+) lymphocyte count and HIV-related symptoms (fever, thrush), it was determined that a yearly decline of more than 300 T cells/microliter of peripheral blood was an independent predictor of the onset of AIDS for subjects with CD4(+) lymphocyte counts of <500 cells/microliter. The results support an important role for failure of T cell homeostasis in the pathogenesis of AIDS. PMID- 9743614 TI - Peripheral human T lymphocyte maintenance of immune functional capacity and phenotypic characteristics following in vivo cocaine exposure. AB - The effects of cocaine exposure upon the host's immune response is equivocal since a variety of studies have generated conflicting conclusions, often as the result of differences between in vitro and/or animal models and the actual conditions experienced in humans who are acutely abusing this drug. To further address this issue, we have studied a group of patients who were positive for cocaine or cocaine metabolites and we evaluated a variety of functional parameters of T-lymphocytes and other peripheral lymphoid cell populations, as well as immunophenotypic characteristics of these cells. When compared to normal controls and patients who were negative for cocaine, we found that the cocaine positive patients had T-cell functional assays which were essentially normal, with the exception of a slight depression in PHA stimulation. Likewise, the immunophenotype of the peripheral blood lymphocytic populations showed normal percentages and numbers of their T cell subsets (CD4, CD8), NK cells, and B cells. Multicolor flow cytometry analysis revealed no difference in T cell subpopulations positive for the "memory" marker, CD62L. No correlation could be established between levels of cocaine or cocaine metabolites and any phenotypic, demographic, or functional parameter. In summary, these results demonstrate that individuals acutely exposed to cocaine do not show markedly altered T cell function or fluctuations in phenotypically identified cell populations. These studies imply that acute cocaine exposure does not predispose individuals to grossly apparent immunosuppression. However, the possibility that subtle, transient, or more specific changes in the immune system may be incurred by use of cocaine, particularly with chronic exposure, remains to be determined. PMID- 9743613 TI - Relationship between plasma IGF-I levels, in vitro correlates of immunity, and human senescence. AB - Insulin-like growth factor-I (IGF-I) is a polypeptide mitogen which is regulated by growth hormone (GH). IGF-I mediates many of the biological functions of GH, including the maintenance of lymphoid mass and functions. Since GH secretion declines with age, we asked whether changes in the availability of IGF-I might contribute to age-associated alterations in immune functions. As a first step, we examined relationships between plasma levels of IGF-I and in vitro correlates of immunity in young and elderly subjects. Heparinized plasma and lymphocytes were collected from the peripheral blood of 34 healthy young (aged 27 +/- 0.9 years, mean +/- SEM) and 41 elderly (79 +/- 1.3 years) volunteers (31 males and 44 females in total). Plasma levels of IGF-I, measured by radioimmunoassay after the removal of IGF-I-binding proteins, were reduced among elders compared to young controls (138 +/- 8.7 ng/mL vs 80.2 +/- 4.7 ng/mL, P < 0.001). The number of circulating lymphocytes did not change with age. The proliferative response ([3H]thymidine uptake into DNA) of T-cells to concanavalin A and B-cells to pokeweed mitogen were reduced among elders (P < 0.05). An increased spontaneous antitumor natural killer (NK) activity (P < 0.001) was accompanied by a higher percentage of CD16(+) NK cells among lymphocytes in older subjects (P < 0.001). The NK cell number was positively related to IGF-I levels in young volunteers but not among elders. Correlation analysis demonstrated a highly significant relationship between plasma IGF-I levels and T-cell (but not B-cell) proliferative response during aging (r = 0.492, P < 0.001). Our results imply that reduced immunocompetence may be one of the consequences of reduced IGF-I levels in human aging. Among the three types of immune cells tested, the T-cells were most sensitive to fluctuations in IGF-I levels. Reduced IGF-I availability may be one of the determinants of the decline in T-cell-mediated immune function in the elderly. To our knowledge, this is the first report presenting correlative data on concurrent changes in IGF-I levels and immune parameters in human aging. PMID- 9743615 TI - Enhancement of antigen-specific IFN-gamma production from CD8(+) T cells by a single amino acid-substituted peptide derived from bovine alphas1-casein. AB - Modulation of CD8(+) T-cell responses specific for an exogenous antigen by epitope variants would be advantageous to develop a novel means of antigen specific immune regulation. We have analyzed CD8(+) T-cell responses to single amino acid-substituted variants of a peptide corresponding to residues 142-149 (p142-149; LAYFYPEL) of alphas1-casein, a major milk allergen, which is a dominant determinant restricted by H-2Kb. An analog peptide L142I with a substitution of Ile for Leu at the nonanchor N-terminal residue induced more IFN gamma secretion than p142-149 from specific CD8(+) T cells. Furthermore, L142I could prime CD8(+) T cells more efficiently in vivo, and these L142I-primed cells secreted more IFN-gamma than p142-149-primed CD8(+) T cells upon stimulation with p142-149 in vitro. These findings are mainly explained by the greater ability of L142I to form stable Kb-peptide complexes. These findings indicate that appropriate analog peptides may be useful as efficient inducers of CD8(+) T cells which recognize the parent peptide and secrete IFN-gamma, a potent inhibitor of Th2-dependent events, including IgE production. PMID- 9743616 TI - Regulation of human lymphocyte IL-4 secretion by intestinal epithelial cell derived interleukin-7 and transforming growth factor-beta. AB - A mucosal immune response to food antigens could result in detrimental hypersensitivity responses. Therefore, the response to many orally administered antigens is downregulated by mechanisms which are not completely understood. Intestinal epithelial cells (IEC) in these tissues may play a role in these regulatory mechanisms via their secreted cytokines. Experiments with human lymphocytes or isolated CD4(+) T cells cultured with 4-day culture supernatants from human colonic carcinoma cell lines revealed that the IEC cell lines normally secreted levels of IL-7 which could enhance IL-4, but not IL-2 or IFN-gamma, secretion by stimulated mixtures of lymphocyes, but not purified CD4(+) T cells. However, acid treatment of the IEC culture supernatants to activate latent TGF beta resulted in a suppression IL-4, but not IL-2 or IFN-gamma, secretion. These results indicate that under normal conditions, IEC secrete latent TGF-beta and IL 7, the latter of which may enhance local IL-4 secretion. However, activation of the IEC-derived TGF-beta may suppress local IL-4 secretion to suppress the induction of local Th2-type responses to intestinal lumenal antigens. PMID- 9743617 TI - Oral tolerance in experimental autoimmune uveoretinitis: feeding after disease induction is less protective than prefeeding. AB - Oral administration of antigen modulates subsequent immune responses raised by conventional subcutaneous priming. If experimental autoantigens are administered, subsequent induction of autoimmune diseases may be inhibited. However, feeding autoantigens after priming or disease induction is more clinically relevant, but the trials have been less successful. Using therapeutic feeding of peptides to inhibit experimental autoimmune uveoretinitis (EAU) induced in LEW rats by bovine S-Ag peptides, we found that only mild disease could be inhibited if feeding was delayed until after immunization, and relatively high feeding doses were required. In recipients with more severe EAU, the clinical efficacy of therapeutic feeding was minimal despite concurrent down-regulation of in vitro antigen-specific lymphocyte proliferation and serum antibody responses. No further inhibition of EAU was found by increasing the feeding dose. Feeding the same peptides prior to immunization produced resistance to moderate to severe disease induction. Unlike prophylactic feeding protocols, conditions were found such that feeding after immunization with low doses of antigen led to worsening of mild disease, raising a note of caution. PMID- 9743618 TI - Abnormal early TCR/CD3-mediated signaling events of a snRNP-autoreactive lupus T cell clone. AB - Multiple immunoregulatory abnormalities characterize systemic lupus erythematosus. Abnormalities of the antigen receptor-mediated early signal transduction biochemical events underscore the diverse cellular aberrations. Fresh peripheral T and B cells and T cell lines from patients with systemic lupus erythematosus display increased Ca2+ responses that are preceded by enhanced antigen receptor-initiated cytosolic protein tyrosine phosphorylation. To further dissect the aberrant signaling events of lupus T cells we studied the early anti CD3 mAb-induced signaling events in autoantigen-specific T cells from lupus patients. We report herein that a lupus snRNP-specific T cell clone, but not other T cells, displays increased Ca2+ fluxes and enhanced production of tyrosine phosphorylated proteins following TCR/CD3 stimulation. PMID- 9743619 TI - Functional analysis of the Escherichia coli genome using the sequence-to structure-to-function paradigm: identification of proteins exhibiting the glutaredoxin/thioredoxin disulfide oxidoreductase activity. AB - The application of an automated method for the screening of protein activity based on the sequence-to-structure-to-function paradigm is presented for the complete Escherichia coli genome. First, the structure of the protein is identified from its sequence using a threading algorithm, which aligns the sequences to the best matching structure in a structural database and extends sequence analysis well beyond the limits of local sequence identity. Then, the active site is identified in the resulting sequence-to-structure alignment using a "fuzzy functional form" (FFF), a three-dimensional descriptor of the active site of a protein. Here, this sequence-to-structure-to-function concept is applied to analysis of the complete E. coli genome, i.e. all E. coli open reading frames (ORFs) are screened for the thiol-disulfide oxidoreductase activity of the glutaredoxin/thioredoxin protein family. We show that the method can identify the active sites in ten sequences that are known to or proposed to exhibit this activity. Furthermore, oxidoreductase activity is predicted in two other sequences that have not been identified previously. This method distinguishes protein pairs with similar active sites from proteins pairs that are just topological cousins, i.e. those having similar global folds, but not necessarily similar active sites. Thus, this method provides a novel approach for extraction of active site and functional information based on three-dimensional structures, rather than simple sequence analysis. Prediction of protein activity is fully automated and easily extendible to new functions. Finally, it is demonstrated here that the method can be applied to complete genome database analysis. PMID- 9743620 TI - GTP-dependent conformational changes associated with the functional switch between Galpha and cross-linking activities in brain-derived tissue transglutaminase. AB - GTP and Ca2+, two well-known modulators of intracellular signaling pathways, control a structural/functional switch between two vital and mutually exclusive activities, cross-linking and Galpha activity, in the same enzyme. The enzyme, a brain-derived tissue-type transglutaminase (TGase), was recently cloned by us in two forms, one of which (s-TGN) lacks a C-terminal region that is present in the other (l-TGN). Immunoreaction with antibodies directed against a peptide present in the C-terminus of l-TGN but missing in s-TGN suggested that this site, which is located in the C-terminal fourth domain, undergoes conformational changes as a result of interaction between l-TGN and GTP. Site-directed mutagenesis suggested that the third domain is involved in mediating the inhibition of the cross linking activity. These results were supported by molecular modeling, which further suggested that domains III and IV both participate in conformational changes leading to the functional switch between the Ca2+-dependent cross-linking activity and the Galpha activity. PMID- 9743621 TI - Regulation of UmuD cleavage: role of the amino-terminal tail. AB - An essential step in SOS mutagenesis is the RecA-mediated posttranslational processing of UmuD-like proteins to the shorter, but mutagenically active, UmuD' like proteins. Interestingly, the UmuD-like proteins undergo posttranslational processing at different rates. For example, although the Escherichia coli UmuD (UmuDEc) and the Salmonella typhimurium UmuD (UmuDSt) proteins are 73% identical, UmuDSt is processed in vivo at a significantly faster rate than the UmuDEc protein. Here, we report experiments aimed at investigating the molecular basis of these phenotypic differences. The faster rate of UmuDSt cleavage probably does not result solely from a better interaction with RecA, since we observed that, in vitro, UmuDSt undergoes RecA-independent autocatalytic processing about four times faster than UmuDEc. By constructing chimeric UmuD proteins, we determined that the amino-terminal tail of the UmuD proteins proximal to the Cys24-Gly25 cleavage site is mainly responsible for the difference in UmuDSt and UmuDEc cleavage rates. Site-directed mutagenesis of the UmuDEc protein suggests that most of the enhanced cleavage observed with the UmuDSt protein can be attributed to the presence of a Pro23 residue, juxtaposed to the cleavage site in UmuDSt. Furthermore, this proline residue appears to result in a UmuD protein that is a much better substrate for intermolecular cleavage. These findings clearly implicate the N-terminal tail of the UmuD-like proteins as playing an important and unexpected regulatory function in the maturation of the mutagenically active UmuD'-like mutagenesis proteins. PMID- 9743622 TI - Affinity, stability and polarity of binding of the TATA binding protein governed by flexure at the TATA Box. AB - The TATA binding protein (TBP), which plays a central role in gene regulation as an essential component of all three nuclear transcription systems, sharply kinks the TATA box at two sites and severely contorts the intervening DNA segment. DNA constructs with precisely localized flexure have been used to investigate the special repertoire of mechanisms and properties that arise from TBP interacting with the TATA box. DNA flexure precisely localized to the sites of TBP-mediated DNA kinking increases the affinity of TBP more than 100-fold; unexpectedly, this increase in affinity is achieved almost exclusively by increasing the stability of the TBP-DNA complex rather than the rate of its formation. In vitro transcription with RNA polymerase III provides a first demonstration that the orientation of TBP on the TATA box is governed by DNA deformability, its C proximal repeat contacting the more flexible end of the TATA box. Exceptionally stable TBP-DNA complexes reach their orientational equilibrium very slowly; in these circumstances, assembly of stable ("committed") transcription initiation complexes can freeze far-from-equilibrium orientations of TBP on the TATA box, causing transcription polarity to be determined by a kinetic trapping mechanism. PMID- 9743623 TI - Identification of phosphate groups involved in metal binding and tertiary interactions in the core of the Neurospora VS ribozyme. AB - We have used ethylation protection experiments and modification interference using phosphorothioate nucleosides to identify phosphate groups involved in the magnesium-dependent tertiary structure and function of the VS ribozyme, a small, self-cleaving RNA. Phosphorothioate interference-rescue experiments in the presence of the thiophilic manganese ion implicate four phosphate groups in direct metal ion binding. Phosphorothioate substitution also creates a new manganese binding site that increases the cis cleavage rate of the ribozyme, possibly by disrupting an inhibitory structure. Interpreting these data in the context of a recently developed structural model shows that almost all of the important phosphate groups are located in the central core of the ribozyme. The model suggests roles for certain phosphate groups in particular steps of RNA folding and identifies a candidate region for the active site of the ribozyme. PMID- 9743624 TI - Isolation and physical characterization of random insertions in Staphylococcal nuclease. AB - Using genetic engineering techniques we generated randomly located internal tandem duplications of random size within Staphylococcal nuclease. Those insertions, possessing greater than 0.1% of normal activity, were sequenced and characterized physically. Insertions were found to begin and end in regions possessing secondary structure as well as in regions without secondary structure. All proteins remained folded and monomeric, although one mutant appeared, by both circular dichroism and size exclusion chromatography, to be partially unfolded. The stability of the insertions as assayed by guanidine hydrochloride denaturation ranged from nearly normal to destabilized by almost 4 kcal per mol. The activities of the insertion mutants ranged from 1/30 to 1/2000 of the parental nuclease. PMID- 9743626 TI - Replication of R6K gamma origin in vitro: discrete start sites for DNA synthesis dependent on pi and its copy-up variants. AB - The regulation of the plasmid R6K gamma origin (gamma ori) is accomplished through the ability of the pi protein to act as an initiator and inhibitor of replication. Hyperactive variants of this protein, called copy-up pi, allow four to tenfold increases of gamma ori plasmid DNA in vivo. The higher activity of copy-up pi variants could be explained by an increase in the initiator function, a decrease in the inhibitor activity, or a derepression of a more efficient mechanism of replication that can be used by wt pi (pi35. 0) only under certain conditions. We have compared the replication activities of wt pi35.0 and copy-up pi mutants in vitro, and analyzed the replication products. It is shown that copy up variants are several-fold more active than wt pi35.0 in replication. This appears to be due to enhanced specific replication activity of copy-up mutants rather than elevated fractions of protein proficient in DNA binding. Furthermore, biochemical complementation revealed that pi200 (copy-up) is dominant over wt pi35.0. The elevated activity of copy-up pi is not caused by an increased rate of replisome assembly as inferred from in vitro replication assays in which the lag periods observed were similar to that of wt pi35.0. Moreover, only one round of semiconservative, unidirectional replication occurred in all the samples analyzed indicating that copy-up pi proteins do not initiate multiple rounds of DNA synthesis. Rather, a larger fraction of DNA template replicates in the presence of copy-up pi as determined by electron microscopy. Two clusters of discrete DNA synthesis start sites are mapped by primer extension near the stability (stb) locus of the gamma ori. We show that the start sites are the same in the presence of wt pi35.0 or copy-up proteins. This comparative analysis suggests that wt pi35.0 and copy-up variants utilize fundamentally similar mechanism(s) of replication priming. PMID- 9743625 TI - Cloning and characterization of a mouse homologue (mNthl1) of Escherichia coli endonuclease III. AB - Endonuclease III (endoIII; nth gene product) of Escherichia coli is known to be a DNA repair enzyme having a relatively broad specificity for damaged pyrimidine bases of DNA. Here, we describe the cloning and characterization of the cDNA and the gene for a mouse homologue (mNthl1/mNth1) of endoIII. The cDNA was cloned from a mouse T-cell cDNA library with a probe prepared by PCR using the library and specific PCR primers synthesized based on the reported information of partial amino acid sequences of bovine NTHL1/NTH1 and of EST Data Bases. The cDNA is 1025 nucleotides long and encodes a protein consisting of 300 amino acids with a predicted molecular mass of 33.6 kDa. The amino acid sequence exhibits significant homologies to those of endoIII and its prokaryotic and eukaryotic homologues. The recombinant mNthl1 with a hexahistidine tag was overexpressed in a nth::cmr nei::Kmr double mutant of E. coli, and purified to apparent homogeneity. The enzyme showed thymine glycol DNA glycosylase, urea DNA glycosylase and AP lyase activities. Northern blot analysis indicated that mNthl1 mRNA is about 1 kb and is expressed ubiquitously. A 15 kb DNA fragment containing the mNthl1 gene was cloned from a mouse genomic library and sequenced. The gene consists of six exons and five introns spanning 6.09 kb. The sequenced 5' flanking region lacks a typical TATA box, but contains a CAAT box and putative binding sites for several transcription factors such as Ets, Sp1, AP-1 and AP-2. The mNthl1 gene was shown to lie immediately adjacent to the tuberous sclerosis 2 (Tsc2) gene in a 5'-to-5' orientation by sequence analysis and was assigned to chromosome 17A3 by in situ hybridization. PMID- 9743627 TI - An arginine residue (Arg101), which is conserved in many GroEL homologues, is required for interactions between the two heptameric rings. AB - Homologous recombination was used to construct a series of hybrid chaperonin genes, containing various lengths of Escherichia coli groEL replaced by the equivalent region from the homologous cpn60-1 gene of Rhizobium leguminosarum. Analysis of proteins produced by these hybrids showed that many of them formed structures with properties consistent with their being single heptameric rings under some conditions, as opposed to the double ring form in which both the GroEL and the Cpn60-1 proteins are found. By determining precise cross-over points, two regions in Cpn60-1 were defined which appeared to be critical for ring-ring interactions. Within one of these regions is a highly conserved arginine residue (Arg101), which we hypothesised to interact with a residue or residues toward the C terminus of the protein, this contact being required for double rings to form. To test this hypothesis, we mutagenised this residue from arginine to threonine in chaperonin genes from two different species of Rhizobium. In both cases, proteins which ran on non-denaturing gels as single rings were produced. Conversion of Arg101 to serine also had the same effect, whereas conversion of Arg101 to lysine did not. Two different single rings created by homologous recombination could be converted back to double rings by changing the threonine, which naturally occurs at this position in E. coli GroEL, back to arginine. The in vivo properties of the proteins were investigated by complementation following deletion of the chromosomal copy of the groEL gene, and by monitoring the ability of cells expressing the hybrid proteins to plate bacteriophage. Most of the hybrid and mutant proteins were functional in these assays, despite their altered properties compared to wild-type GroEL. PMID- 9743628 TI - Three-dimensional folding of an RNA hairpin required for packaging HIV-1. AB - An NMR-based structure is presented for a 20 mer hairpin model of the SL3 stem loop from the HIV-1 packaging signal. The stem has an A-family structure. However, the GGAG tetraloop appears to be flexible with the second (G10) and fourth (G12) bases extruded from the normal stacking arrangement. The A-base (A11) occupies a cavity large enough for it to jump rapidly between stacking upon G9 (in the loop) and G13 (from the base-pair adjacent to the loop). The H-bonding loci of G10, A11, and G12 are unoccupied in the free RNA structure. The loop should be easily adaptable to binding by the HIV-1 nucleocapsid protein or loop receptors. PMID- 9743629 TI - Are the light-harvesting I complexes from Rhodospirillum rubrum arranged around the reaction centre in a square geometry? AB - The basic photosynthetic unit containing the reaction centre and the light harvesting I complex (RC-LHI) of the purple non-sulphur bacterium Rhodospirillum rubrum was purified and reconstituted into two-dimensional (2D) membrane crystals. Transmission electron microscopy using conventional techniques and cryoelectron microscopy of the purified single particles and of 2D crystals yielded a projection of the RC-LHI complex at a resolution of at least 1.6 nm. In this projection the LHI ring appears to have a square symmetry and packs in a square crystal lattice. The square geometry of the LHI ring was observed also in images of single isolated particles of the RC-LHI complex. However, although the LHI units are packed identically within the crystal lattice, a new rotational analysis developed here showed that the reaction centres take up one of four possible orientations within the ring. This fourfold disorder supports our interpretation of a square ring symmetry and suggests that a hitherto undetected component may be present within the photosynthetic unit. PMID- 9743630 TI - Projection structures of three photosynthetic complexes from Rhodobacter sphaeroides: LH2 at 6 A, LH1 and RC-LH1 at 25 A. AB - Three photosynthetic complexes, light-harvesting complex 2 (LH2), light harvesting complex 1 (LH1), and the reaction centre-light-harvesting complex 1 photounit (RC-LH1), were purified from a single species of a purple bacterium, Rhodobacter sphaeroides, and reconstituted into two-dimensional (2-D) crystals. Vesicular 2-D crystals of LH1 and RC-LH1 were imaged in negative stain and projection maps at 25 A resolution were produced. The rings formed by LH1 have approximately the same mean diameter as the LH1 rings from Rhodospirillum rubrum ( approximately 90 A) and therefore are likely to be composed of 15 to 17 alphabeta subunits. In the projection map calculated from the RC-LH1 2-D crystals, the reaction centre is represented by an additional density in the centre of the ring formed by the LH1 subunits. The marked improvement of shape and fine structure after a rotational pre-alignment of the RC-LH1 unit cells before averaging strongly suggests that the RC is not in a unique orientation within the LH1 rings. Tubular crystals of LH2 showed a high degree of order and allowed calculation of a projection map at 6 A resolution from glucose-embedded specimens. The projection structure shows a ring of nine alphabeta subunits. Variation of the alpha-helical projection densities suggests that the 9-fold symmetry axis is tilted with respect to the membrane normal. PMID- 9743631 TI - Water molecules in DNA recognition I: hydration lifetimes of trp operator DNA in solution measured by NMR spectroscopy. AB - The present NMR study investigates the residence times of the hydration water molecules associated with uncomplexed trp operator DNA in solution by measuring intermolecular nuclear Overhauser effects (NOE) between water and DNA protons, and the nuclear magnetic relaxation dispersion (NMRD) of the water 2H and 17O resonances. Both methods indicate that the hydration water molecules exchange with bulk water on the sub-nanosecond time scale at 4 degreesC. No evidence was obtained for water molecules bound with longer residence times. In particular, the water molecules at the sites of interfacial hydration in the trp repressor/operator complex do not seem kinetically stabilized in the uncomplexed DNA. Analysis of the crystal structures of two different trp repressor/operator complexes shows very similar structural environments for the water molecules mediating specific contacts between the protein and the DNA, whereas much larger variations are observed for the location of corresponding water molecules detected in the crystal structure of an uncomplexed trp operator DNA duplex. Therefore, it appears unlikely that the hydration characteristics of the uncomplexed DNA target would be a major determinant of trp repressor/operator recognition. PMID- 9743632 TI - Water molecules in DNA recognition II: a molecular dynamics view of the structure and hydration of the trp operator. AB - The structure and hydration of the DNA duplex d-(AGCGTACTAGTACGCT)2 corresponding to the trp operator fragment used in the crystal structure of the half site complex (PDB entry 1TRR) was studied by a 1.4 ns molecular dynamics simulation in water. The simulation, starting from a B-DNA conformation, used a non-bonded cutoff of 1.4 nm with a reaction field correction and resulted in a stable trajectory. The average DNA conformation obtained was closer to the ones found in the crystal structures of the complexes (PDB entries 1TRO and 1TRR) than to the crystal structure of unbound trp operator (Nucleic Acid Database entry BDJ061). The DNA hydration was characterized in terms of hydrogen bond percentages and corresponding residence times. The residence times of water molecules within 0.35 nm of the DNA non-exchangeable protons were calculated for comparison with NMR measurements of intermolecular water-DNA NOEs and nuclear magnetic relaxation dispersion measurements. No significant difference was found between major and minor groove hydration. The DNA donors and acceptors were hydrogen bonded to water molecules for 77(+/-19)% of the time on average. The average residence time of the hydrogen bonded water molecules was 11(+/-11) ps with a maximum of 223 ps. When all water molecules within NOE distance (0.35 nm) of non-exchangeable protons were considered, the average residence times increased to an average of 100(+/-4) ps and a maximum of 608 ps. These results agree with the experimental NMR results of Sunnerhagen et al. which did not show any evidence for water molecules bound with more than 1 ns residence time on the DNA surface. The exchange of hydration water from the DNA occurred in the major groove primarily through direct exchange with the bulk solvent, while access to and from the minor groove frequently proceeded via pathways involving ribose O3' and O4' and phosphate O2P oxygen atoms. The most common water diffusion pathways in the minor groove were perpendicular to the groove direction. In general, water molecules visited only a limited number of sites in the DNA grooves before exiting. The hydrogen bonding sites, where hydrogen bonds could be formed with donor and acceptor groups of the DNA, were filled with water molecules with an average B factor value of 0.58 mn2. No special values were observed at any of the sites, where water molecules were observed both in the trp repressor/operator co crystals and in the crystal structure of unbound DNA. PMID- 9743633 TI - Structures of free and complexed forms of Escherichia coli xanthine-guanine phosphoribosyltransferase. AB - Structures of free, substrate-bound and product-bound forms of Escherichia coli xanthine-guanine phosphoribosyltransferase (XGPRT) have been determined by X-ray crystallography. These are compared with the previously determined structure of magnesium and sulphate-bound XPRT. The structure of free XGPRT at 2.25 A resolution confirms the flexibility of residues in and around a mobile loop identified in other PRTases and shows that the cis-peptide conformation of Arg37 at the active site is maintained in the absence of bound ligands. The structures of XGPRT complexed with the purine base substrates guanine or xanthine in combination with cPRib-PP, an analog of the second substrate PRib-PP, have been solved to 2.0 A resolution. In these two structures the disordered phosphate binding loop of uncomplexed XGPRT becomes ordered through interactions with the 5'-phosphate group of cPRib-PP. The cyclopentane ring of cPRib-PP has the C3 exo pucker conformation, stabilised by the cPRib-PP-bound Mg2+. The purine base specificity of XGPRT appears to be due to water-mediated interactions between the 2-exocyclic groups of guanine or xanthine and side-chains of Glu136 and Asp140, as well as the main-chain oxygen atom of Ile135. Asp92, together with Lys115, could help stabilise the N7-protonated tautomer of the incoming base and could act as a general base to remove the proton from N7 when the nucleotide product is formed. The 2.6 A resolution structure of XGPRT complexed with product GMP is similar to the substrate-bound complexes. However, the ribose ring of GMP is rotated by approximately 24 degrees compared with the equivalent ring in cPRib PP. This rotation results in the loss of all interactions between the ribosyl group and the enzyme in the product complex. PMID- 9743635 TI - Supersites within superfolds. Binding site similarity in the absence of homology. AB - A method is presented to assess the significance of binding site similarities within superimposed protein three-dimensional (3D) structures and applied to all similar structures in the Protein Data Bank. For similarities between 3D structures lacking significant sequence similarity, the important distinction was made between remote homology (an ancient common ancestor) and analogy (likely convergence to a folding motif) according to the structural classification of proteins (SCOP) database. Supersites were defined as structural locations on groups of analogous proteins (i.e. superfolds) showing a statistically significant tendency to bind substrates despite little evidence of a common ancestor for the proteins considered. We identify three potentially new superfolds containing supersites: ferredoxin-like folds, four-helical bundles and double-stranded beta helices. In addition, the method quantifies binding site similarities within homologous proteins and previously identified supersites such as that found in the beta/alpha (TIM) barrels. For the nine superfolds, the accuracy of predictions of binding site locations is assessed. Implications for protein evolution, and the prediction of protein function either through fold recognition or tertiary structure comparison, are discussed. PMID- 9743636 TI - Intangible rewards of dentistry. PMID- 9743634 TI - Temperature effects on the allosteric responses of native and chimeric aspartate transcarbamoylases. AB - Although structurally very similar, the aspartate transcarbamoylases (ATCase) of Serratia marcescens and Escherichia coli have distinct allosteric regulatory patterns. It has been reported that a S. marcescens chimera, SM : rS5'ec, in which five divergent residues (r93 to r97) of the regulatory polypeptide were replaced with their Escherichia coli counterparts, possessed E. coli-like regulatory characteristics. The reverse chimera EC:rS5'sm, in which the same five residues of E. coli have been replaced with their S. marcescens counterpart, lost both heterotrophic and homotropic responses. These results indicate that the r93 r97 region is critical in defining the ATCase allosteric character. Molecular modeling of the regulatory polypeptides has suggested that the replacement of the S5' beta-strand resulted in disruption of the allosteric-zinc interface. However, the structure-function relationship could be indirect, and the disruption of the interface could influence allostery by altering the global energy of the enzyme. Studies of the temperature-sensitivity of the CTP response demonstrate that it is possible to convert CTP inhibition of the SM:rS5'ec chimera at high temperature to activation below 10 degreesC. Nonetheless, the temperature response of the native S. marcescens ATCase suggests a strong entropic effect that counteracts the CTP activation. Therefore, it is suggested that the entropy component of the coupling free energy plays a significant role in the determination of both the nature and magnitude of the allosteric effect in ATCase. PMID- 9743637 TI - Three-dimensional guidance system for implant insertion: Part I. AB - A technique is presented, with two sets of newly designed drills, that facilitates three-dimensional guided insertion of implants. A radiographic guide has been fabricated with titanium markers that are transferred into the CT scan reformatted images. The resulting cross-sectional and panoramic reformatted images provide the precise location for each implant. The buccolingual and mesiodistal inclinations of each implant are measured and transferred to a surgical guide that contains steel drill guide tubes. The starting point and three-dimensional orientation of each implant is then transferred to a pilot osteotomy. The widening of an osteotomy, with end-cutting drills of the usual design, can lead to the loss of orientation because a wider drill cannot be effectively guided by a smaller diameter osteotomy. A newly designed set of drills has been developed that maintains tracking of the original orientation as the incremental increase in width proceeds. The three-dimensional guidance system for implant insertion was first accomplished in vitro and then on patients. PMID- 9743638 TI - Reconstruction of advanced mandibular resorption with both subperiosteal and root form implants. AB - A combination implant reconstruction using both subperiosteal and endosseous root form implants for advanced mandibular resorption is presented. In theory and practice, physiologic endosseous implant support should prevent further resorption in the mandibular symphysis region. Three patients' post-treatment outcomes demonstrate a complete return of mandibular oral function. The greater risk of mandibular fracture, which is associated with other treatment options (multiple root-form implants or transmandibular implants), is avoided with this treatment approach. With follow-up periods ranging from 6 months to 28 months the three patients are satisfied with comfort, function, and appearance. A long-term follow-up study is planned for future publication. PMID- 9743639 TI - Morphological and immunohistochemical aspects of the biological seal in Klockner's dental implants: study of 15 cases. AB - The morphological and immunohistochemical aspects of the peri-implanted pathological soft tissue of 21 Klockner implants are assessed, corresponding to 15 patients and three gingival samples from three cadavers. For the collagen and vascular walls assessment, the Picro-Sirio technique is used with normal microscopy as well as with polarized light, evaluating the decrease of collagen fibers in relation to the grade of inflammation. In cases when there is a plasmocyte inflammatory prevalence, antibodies are used contrasted with Kappa and Lambda light chains, observing in all cases a polyclonal plasmocytosis (reactive). The Ag. Ki-67 is used to mark cell proliferation that shows a normal or enlarged activity that can reach the infiltrated cells of the lamina propria. Epitheliotropism or a lymphoepithelial lesion is analyzed in relation to the inflammation, observing the tendency to be larger in K-type implants with a completely-submerged technique. The lesion that is usually observed is the periimplanted mucositis of the chronic or mixed type. The regenerated epithelium has a slight thickness with congestive and dilated vessels. In one case (as anecdotal value), there is the presence of coilocytes compatible with infection by HPV (Human Papilloma Virus). PMID- 9743640 TI - A multipurpose template for implant placement. AB - A case report is presented describing the laboratory fabrication and clinical use of an innovative template. This template was designed to function as a radiographic implant positioning guide, accurate bone sounding guide, and surgical guide. This appliance is intended to serve additional roles as an aid in flap reflection and as a pick-up impression tray. PMID- 9743642 TI - Prevalence of malocclusion and orthodontic treatment need in the United States: estimates from the NHANES III survey. AB - Data from the third National Health and Nutrition Examination Survey (NHANES III) provide a clear picture of malocclusion in the US population. Noticeable incisor irregularity occurs in the majority of all racial/ethnic groups, with only 35% of adults having well-aligned mandibular incisors. Irregularity is severe enough in 15% that both social acceptability and function could be affected, and major arch expansion or extraction of some teeth would be required for correction. About 20% of the population have deviations from the ideal bite relationship; in 2% these are severe enough to be disfiguring and are at the limit for orthodontic correction. In Mexican-Americans compared to the rest of the population, incisor irregularity and both severe Class II and Class III malocclusions are more prevalent, but deep bite and open bite are less prevalent. Application of the Index of Treatment Need to the survey data reveals that 57% to 59% of each racial/ethnic group has at least some degree of orthodontic treatment need. Over 30% of white youths, 11% of Mexican-Americans, and 8% of blacks report receiving treatment. Severe malocclusion is observed more frequently among blacks, which may reflect their lower level of treatment. Treatment is much more frequent in higher income groups, but approximately 5% of those in the lowest income group and 10% to 15% of those in intermediate income groups report being treated. PMID- 9743641 TI - Planning the transition to a comprehensive implant rehabilitation: a case report- Part I. AB - When the edentulous condition has existed for many years, extensive ridge resorption may complicate the ability to function with a conventional prosthesis. Implant therapy in conjunction with bone augmentation procedures may provide new options for the predictable restoration of the edentulous state. However, complex implant reconstruction may place considerable stress on the patient's ability to function throughout therapy. Providing stable interim prostheses and smooth transitions between procedures may alleviate the problem. This can be accomplished only with extensive diagnosis, treatment planning, and the judicious use of transitional prostheses. The first part of this case report focuses on the planning stages of a complex implant dental rehabilitation. PMID- 9743643 TI - Expectations and perceptions regarding treatment: a prospective study of patients undergoing orthognathic surgery. AB - One hundred consecutive patients undergoing orthognathic surgical treatment were interviewed concerning their problems before surgery, motives for seeking treatment, and the effects of treatment 1 year after surgery. Comparison with other studies was undertaken using questionnaires validated in such studies. Before surgery, problems relating to function were most significant, followed by esthetic concerns and, to a far lesser extent, social interaction-type problems. Motives for seeking treatment also related mostly to functional issues. Such issues were considered best resolved through treatment. The finding that functional issues were of greater significance to patients than esthetic concerns differs from findings in most previous studies. The difference may at least partly be explained by sociocultural differences. PMID- 9743644 TI - A comparison of patients requiring orthognathic treatment who subsequently refused or accepted surgery. AB - Forty-four patients who required combined orthodontic-surgical procedures were psychologically assessed in relation to stress, self-esteem, introspectiveness, and body satisfaction. Thirty patients accepted the surgical procedures, and 14 refused surgery. These two groups of patients were compared with respect to their psychologic profiles; statistical analysis detected no significant differences. PMID- 9743645 TI - Postoperative migration of the osteotomy segment stabilized by titanium miniplate osteosynthesis following Le Fort I osteotomy: an x-ray stereometric study. AB - The aim of the present study was to evaluate whether titanium miniplate fixation prevents postoperative migration of the osteotomy segment following Le Fort I osteotomy and, if not, to evaluate whether postoperative migration varies with the direction and amount of surgical repositioning. In 10 consecutive patients who received Le Fort I osteotomies, postoperative migration of the osteotomy segment was studied in a three-dimensional system by means of x-ray stereometry. Surgical repositioning was recorded by cephalometry in lateral head films obtained before surgery and at 2 days postoperatively. X-ray stereograms were obtained at intervals from 2 days to 1 year after surgery. During the 1-year observation period, significant postoperative migration of the osteotomy segment was found at some stage in nine patients. No correlation between the amount of surgical repositioning and postoperative migration was found. However, a tendency toward superiorly directed postoperative migration of the osteotomy segment, independent of the direction of surgical repositioning, was observed. It is thus concluded that titanium miniplates do not prevent postoperative migration of the osteotomy segment. The weak correlation between the amount of surgical repositioning and postoperative migration of the osteotomy segment indicates that predictions of individual outcomes of surgical corrections are uncertain. PMID- 9743646 TI - Stability after inferior or anterior maxillary repositioning by Le Fort I osteotomy: a biplanar stereocephalometric study. AB - A biplanar cephalometric stereoradiographic system was used to measure maxillary positional changes during and after Le Fort I procedures for inferior or anterior repositioning of the maxilla. Of the 29 patients studied, 25 had undergone surgical maxillary advancement, and 15 had undergone inferior repositioning. These cases were divided into subgroups based on fixation technique, presence or absence of bone grafts, age, and surgeon. The postsurgical follow-up period was at least 11 months for each patient. The landmarks used to measure maxillary positional changes consisted of skeletal features and points on wire or rigid fixation devices. Mean magnitudes of postsurgical maxillary landmark displacement were insignificant for all subgroups examined except the advancement group with wire fixation. Individual patients exhibiting the greatest postsurgical displacements tended to have had large surgical advancements and received wire fixation. Magnitudes of surgical and postsurgical change did not seem correlated on the whole, except in the maxillary advancement cases in which bone grafts had not been used. Error analysis results suggest that digitizing consistency was acceptable for maxillary landmarks; however, methodologic improvements with regard to provision of cranial reference landmarks may further minimize errors in future studies. PMID- 9743647 TI - Relationship between disc displacement and morphologic features of skeletal Class III malocclusion. AB - The present study assessed the relationship between disc displacement and facial morphologic features in 48 patients with skeletal Class III malocclusion. Clinical examination, standardized lateral oblique temporomandibular joint radiographs, and cephalographs were used to investigate this relationship. Disc displacement was evaluated by the presence of joint sounds (clicking and crepitation) or restricted condylar translation at maximal mouth opening. Thirty two patients with signs of disc displacement were compared with 16 patients without signs of disc displacement. Results showed that patients with signs of disc displacement had a significantly larger gonial angle and/or SN-MP of the mandible. Thus, morphologic features of patients with skeletal Class III malocclusion may represent a risk factor for the development of disc displacement. PMID- 9743648 TI - Simultaneous rhinoplasty and maxillomandibular osteotomies: Indications and contraindications. AB - Functional and esthetic correction of a dentofacial deformity may require not only maxillary or mandibular osteotomies but also a rhinoplasty. Rigid internal fixation makes it possible to perform rhinoplasty and maxillary osteotomies simultaneously. Nevertheless, to plan rhinoplasty correctly it is of paramount importance to predict changes of the nose that will follow osteotomy of the maxilla. The authors present their experience concerning the surgical technique, advantages, and disadvantages of rhinoplasty in combination with orthognathic surgery. PMID- 9743649 TI - Use of selectively colored stereolithography for diagnosis of impacted supernumerary teeth for a patient with cleidocranial dysplasia. AB - The objective of the present study was to develop a three-dimensional method of locating the position of impacted supernumerary teeth in a patient with cleidocranial dysplasia. Stereolithographies of the patient's teeth and maxillofacial structure were prepared, coloring the teeth red. This model established accurate tooth malposition and provided support for selection of surgical exposure for extraction and traction of the impacted teeth. Selectively colored stereolithography is expected to be useful in a number of diverse situations. PMID- 9743650 TI - Laboratory evaluation of adhesively crimped surgical ball hooks. AB - The aim of this study was to examine the effect of the addition of sandblasting and/or dental adhesive on the stability of the crimpable hook when positioned and crimped onto surgical arch wires. Ninety crimpable ball hooks were divided into six test groups: crimp only; apply Panavia 21 and crimp; apply C & B Metabond and crimp; sandblast and crimp; sandblast, apply Panavia 21, and crimp; and sandblast, apply C & B Metabond, and crimp. Each hook was treated according to the criteria of the relevant test group and then crimped to the arch wire. The force required to dislodge each hook from the arch wire was measured. The results demonstrated that sandblasting caused a significant increase in the force required to dislodge the crimped hook. The addition of either Panavia 21 or C & B Metabond adhesives also resulted in a significant increase in the required dislodging force. The force required to dislodge the hook was increased by a factor of 10 where sandblasting and Panavia 21 were applied. The same increase was observed where C & B Metabond was applied, without sandblasting. However, it was concluded that the use of Panavia 21, together with an intraoral sandblasting machine, would be more appropriate in the clinical setting, primarily due to the ease of use associated with Panavia 21. PMID- 9743651 TI - The publishing process. PMID- 9743652 TI - Treatment of medial and lateral epicondylitis--tennis and golfer's elbow--with low level laser therapy: a multicenter double blind, placebo-controlled clinical study on 324 patients. AB - BACKGROUND AND OBJECTIVE: Among the other treatment modalities of medial and lateral epicondylitis, low level laser therapy (LLLT) has been promoted as a highly successful method. The aim of this clinical study was to assess the efficacy of LLLT using trigger points (TPs) and scanner application techniques under placebo-controlled conditions. STUDY DESIGN/MATERIAL AND METHODS: The current clinical study was completed at two Laser Centers (Locarno, Switzerland and Opatija, Croatia) as a double-blind, placebo controlled, crossover clinical study. The patient population (n = 324), with either medial epicondylitis (Golfer's elbow; n = 50) or lateral epicondylitis (Tennis elbow; n = 274), was recruited. Unilateral cases of either type of epicondylitis (n = 283) were randomly allocated to one of three treatment groups according to the LLLT technique applied: (1) Trigger points; (2) Scanner; (3) Combination Treatment (i.e., TPs and scanner technique). Bilateral cases of either type of epicondylitis (n = 41) were subject to crossover, placebo-controlled conditions. Laser devices used to perform these treatments were infrared (IR) diode laser (GaAlAs) 830 nm continuous wave for treatment of TPs and HeNe 632.8 nm combined with IR diode laser 904 nm, pulsed wave for scanner technique. Energy doses were equally controlled and measured in Joules/cm2 either during TPs or scanner technique sessions in all groups of patients. The treatment outcome (pain relief and functional ability) was observed and measured according to the following methods: (1) short form of McGill's Pain Questionnaire (SF-MPQ); (2) visual analogue scales (VAS); (3) verbal rating scales (VRS); (4) patient's pain diary; and (5) hand dynamometer. RESULTS: Total relief of the pain with consequently improved functional ability was achieved in 82% of acute and 66% of chronic cases, all of which were treated by combination of TPs and scanner technique. CONCLUSIONS: This clinical study has demonstrated that the best results are obtained using combination treatment (i.e., TPs and scanner technique). Good results are obtained from adequate treatment technique correctly applied, individual energy doses, adequate medical education, clinical experience, and correct approach of laser therapists. We observed that under- and overirradiation dosage can result in the absence of positive therapy effects or even opposite, negative (e.g., inhibitory) effects. The current clinical study provides further evidence of the efficacy of LLLT in the management of lateral and medial epicondylitis. PMID- 9743653 TI - Intrapulpal temperature during preparation with the Er:YAG laser compared to the conventional burr: an in vitro study. AB - OBJECTIVE: The authors explored whether the preparation with the Er:YAG laser showed a difference in increase of intrapulpal temperature in comparison to the conventional burr. SUMMARY BACKGROUND DATA: The effect of preparation with the Er:YAG laser on the temperature in the pulp is probably the biggest problem in using the laser for preparation of dental hard tissue. METHODS: The effect on the temperature in the pulp was studied on extracted human incisors and canines in vitro in palatinal class I cavities. The temperature during preparation with the Er:YAG laser was compared to that recorded during conventional tactile preparation with a diamond burr. The study was designed so that the pulpal cavity and the measuring probe were kept at a constant temperature of 37 degrees C from the root upward while the crown, which was thermally isolated, was exposed to preparation and cooling agents. RESULTS: During preparation with the laser, there was a temperature drop after a few seconds from 37 degrees C to 25 degrees C to 30 degrees C due to cooling with water and air. Even with trepanation, there was only an increase in temperature in the pulp when the temperature measuring probe was hit directly by the laser beam. With conventional preparation in comparison, even before trepanation there was a rise in temperature to more than 60 degrees C. CONCLUSIONS: The reduction in pain with clinical use of the Er:YAG laser for class V cavities has already been mentioned in publications and could, in addition to the nontactile preparation, be due to the lesser increase in intrapulpal temperature during the laser preparation in comparison to the conventional burr. PMID- 9743655 TI - Morphological and atomic analytical changes after CO2 laser irradiation emitted at 9.3 microns on human dental hard tissues. AB - OBJECTIVE: The purposes of this study were to examine the effects of CO2 laser emitted at 9.3 microns on human sound and carious dental hard tissue ablation with a stereoscope, scanning electron microscope (SEM), and energy dispersive X ray spectrometer (SEM-EDX) and to identify possible applications of this laser in clinical treatment. SUMMARY BACKGROUND DATA: There has been no report of morphological changes or atomic analytical studies on carious hard tissues after laser irradiation with 9.3 microns CO2 laser. METHODS: Sixty extracted human teeth with no caries and sixty teeth with enamel or dentin caries were used for this study. All teeth were horizontally sectioned into slices (approximately 3 mm in thickness) and the samples were irradiated with CO2 laser using the following two parameters: a fluence of 78 J/cm2 and 5 pps for 2 sec. After laser irradiation, half of the samples were observed by stereoscopy and SEM and the other half were analyzed by SEM-EDX. RESULTS: The lased sound enamel and dentin surfaces showed crater-like structures which had been produced by the high laser energy. On the other hand, some portions of carious hard tissues were evaported by the laser. A slight amount of carbonization was observed by stereoscopy. Calcium (Ca) and phosphorus (P) content of sound or carious hard tissues was increased significantly (p < 0.01) after laser irradiation, but the ratio of Ca to P after laser irradiation was significantly increased (p < 0.01) on sound hard tissue only. CONCLUSION: These results suggest that the 9.3 microns CO2 laser may be useful for the prevention or removal of caries in clinical situations. PMID- 9743654 TI - Import of radiation phenomena of electrons and therapeutic low-level laser in regard to the mitochondrial energy transfer. AB - OBJECTIVE: The authors describe a consistent theoretical model of the cellular energy transfer (respiratory chain) by taking into consideration the radiation phenomena of electrons and therapeutic low level laser. SUMMARY BACKGROUND DATA: Biochemical models of the cellular energy transfer regard the classical corpuscular aspect of electrons as the responsible energy carriers, thereby ignoring the wave-particle dualism of the electrons and the import of radiation energy in this process. METHODS: The authors show the influence of radiation phenomena on the cellular energy transfer, explaining consistently some of the intermediate steps of this complex process. RESULTS: Because of the inherent wave particle dualism of the electrons, it is appropriate to regard radiation phenomena to explain the cellular energy transfer. The classical biochemical models use only the particle part of the electrons as energy carriers. The connection between energy transport by radiation and the order in structures may be understood if, for instance, structurally bound energy is released during the dissolution of structures (oxidation of foodstuffs) or is again manifested (final reduction of oxygen to water). With a attention to the energy values relevant for the respiratory chain, the import of electromagnetic radiation of characteristic ranges of wavelengths on the cellular energy transfer becomes evident. Depending on its wavelength, electromagnetic radiation in the form of light can stimulate macromolecules and can initiate conformation changes in proteins or can transfer energy to electrons. Low level laser from the red and the near infrared region corresponds well with the characteristic energy and absorption levels of the relevant components of the respiratory chain. This laser stimulation vitalizes the cell by increasing the mitochondrial ATP(adenosine-tri-phosphate)-production. CONCLUSIONS: With regard to radiation phenomena and its inhanced electron flow in the cellular energy transfer (respiratory chain), it is possible to explain the experimentally found increase of ATP-production by means of low-level laser light on a cellular level. Intense research for this biostimulative effect is still necessary. PMID- 9743656 TI - Design and development of an electro-optic measurement system for 3-D topographic surveys: application to skin nevoid lesions. AB - OBJECTIVE: The authors designed and developed a system for the measurement of skin lesions geometrical proprieties. The measured characteristics can be of extreme importance in order to provide elements for the early diagnosis of melanoma. SUMMARY BACKGROUND DATA: Actually, medical experience and prevention are the only instruments for melanoma diagnosis. Some studies have been conducted with thermocameras and with ecography and photoacoustics spectrography. METHODS: A scanning laser-based measurement system for non-contact shape measurement has been designed and tested. The system measurement characteristics have been studied to assure an accurate reconstruction of skin lesion surface characteristics. The laser sensor is fixed on a motorized system that scans the lesion surface. All the controls on the scanning system and on the recording of data have been automated and are software-controlled. RESULTS: Full success in the measurement and reconstruction of the skin lesions characteristics has been reached. An accuracy of 0.03 mm has been obtained with the system. Measurements of the height and perimeter of many suspected lesions have been performed. CONCLUSIONS: The research reported in this work has demonstrated the applicability and usefulness of the proposed method by means of an in-vitro prototype-version measurement system. The possibility of surveying the shape of skin lesions by means of a non-contact electro-optic measurement plays an important role in the objective evaluation of in vivo pigmented suspected nevoid lesions (melanoma). Next steps include the study of a possible correlation between topographic elements and histological tests and the possibility of controlling suspected lesions employing this technique both in the diagnostic and the follow-up phases. PMID- 9743657 TI - Surgical cleansing of varicose ulcers of the leg using a CO2 laser with rotating mirror scanner. AB - OBJECTIVE: Our purpose was to evaluate the effectiveness of cleansing by CO2 laser with a rotating mirror scanner in accelerating ulcer granulation and reducing sensitization, bleeding, and the need for painful medications. SUMMARY BACKGROUND DATA: Cleansing, the first step in treating phlebostatic ulcers, may be performed with a defocussed CO2 laser, but laser-to-target distance, technique, and duration of the procedure may vary. METHODS: We studied 20 patients, ages 38-85, with "common" phlebostatic ulcers showing a tendency not to granulate. The patients were divided into two groups by sex (7 females and 3 males) and by size of ulcer. The first group was treated using the CO2 laser and rotating mirror scanner; the second was treated surgically with scissors and tweezers. In both groups debridement was followed by mobile and fixed elastocompressing bandage, mobilization, and phlebotonics. Six patients in the first group and four in the second subsequently underwent short stripping to correct the incompetent saphenous circulation. RESULTS: Ulcer cleansing was achieved in all 20 cases. In the laser group, the treatment was well tolerated by 9 patients and granulation was achieved in 10 days on average; in the second group, cleansing was very painful in 5 cases and granulation was achieved in 25 days on average. Both groups had good cicatrization at 3 months after elastocompression, zinc dioxide medication, mobilization, and phlebotonics. The 6 month follow-up revealed only one recurrence--a small ulcer at a different location--in the first group and no cases of recurrence in the second. CONCLUSIONS: Cleansing by CO2 laser and rotating mirror scanner allows the acceleration of ulcer granulation and reduces sensitization, bleeding, and the need for painful and costly medications. PMID- 9743658 TI - Where have all the applicants gone? PMID- 9743659 TI - The state of dental research: a time for change. PMID- 9743661 TI - Palatogram assessment of maxillary complete dentures. AB - Phonetics, esthetics, function, and comfort form the foundation of a successful dental prosthesis. A review of the mechanics of speech as well as common speech problems encountered with a removable maxillary prosthesis are presented. The use of a palatogram to aid the clinician in the assessment and resolution of speech problems associated with a maxillary denture is demonstrated. PMID- 9743660 TI - The management of implant-retained overdenture treatment with custom-made metallic attachment housing. AB - A technique using a custom-made cast alloy attachment housing for an implant retained overdenture prosthesis is presented. Common problems encountered in implant-tissue bar-retained overdentures are discussed. Details of the modified laboratory procedure are described. This technique provides the clinician with an alternative for predictably treating patients with implant-retained overdentures. PMID- 9743662 TI - Evaluation of working distances at a 1:1 reproduction ratio for seven popular 35 mm dental camera systems. PMID- 9743663 TI - Marginal adaptation of implant-supported metal-ceramic crowns fabricated with gold cylinders. AB - PURPOSE: The purpose of this investigation was to determine the mean marginal discrepancy of metal-ceramic crowns fabricated with gold cylinders and cemented on implant abutments. These discrepancies were then compared with those measured previously for implant-supported ceramic crowns. MATERIALS AND METHODS: Fifteen Nobel BioCare CeraOne abutments were connected to implant fixtures embedded in acrylic resin blocks. Marginal discrepancies were determined for gold cylinders, gold cylinders plus ceramic alloy (metal frameworks), completed metal-ceramic crowns, and cemented metal-ceramic crowns using a stereomicroscope equipped with a video camera linked to a computer. A Hotelling's T2 test (p < or = .05) was used to evaluate potential differences in mean marginal discrepancies among groups. RESULTS: The mean marginal discrepancies were: 1) gold cylinders, 7.56 +/ 2.73 microns; 2) metal frameworks, 6.21 +/- 1.34 microns; 3) metal-ceramic crowns, 11.06 +/- 3.21 microns; and 4) zinc-phosphate cemented crowns, 31.47 +/- 6.65 microns. No significant difference between gold cylinders and metal frameworks was found. Mean marginal discrepancies for metal-ceramic crowns were significantly greater than discrepancies for cast gold cylinders. Cemented-crown mean marginal discrepancy was significantly greater than all other means. CONCLUSIONS: Cemented metal-ceramic crowns fabricated using proprietary gold cylinders exhibited well-fitting margins (31.47 microns). PMID- 9743664 TI - The effects of custom tray material on the accuracy of master casts. AB - PURPOSE: The accuracy of master cast reproduction by a polyvinylsiloxane impression material using two visible-light-curing resin and an autopolymerizing polymethyl methacrylate resin custom tray materials was investigated. MATERIALS AND METHODS: Custom trays were fabricated from a master cast that had three index points marked on both the inner and outer vestibules. Impressions were made of the master cast using Extrude and then poured in Die Keen Green stone. The distances between the reproduced index points were measured to +/- 0.01 mm with a traveling microscope and the algebraic norms calculated for each tray material. RESULTS: No differences (p > .05) were found in the dimensions of the inner index points, while the separations of the outer index points indicated that there were differences in the accuracy of reproduction (p < .01) by the three tray materials. The index points reproduced on the casts, compared with the master cast, were closer together for the Triad Blue trays than for the TruTray and Ontray impression trays. CONCLUSIONS: All three tray materials produced acceptable casts, but the greatest accuracy was achieved with TruTray, followed by Ontray. Triad Blue produced casts that were slightly smaller than the master. In practice, the small measured differences in cast dimensions may not have clinical significance. PMID- 9743665 TI - Tear strength of four irreversible hydrocolloid impression materials. AB - PURPOSE: The purpose of this investigation was to examine the tear strength of four irreversible hydrocolloid (alginate) impression materials (Tare-Free Alg, Jeltrate, Identic, and Kromopan). MATERIALS AND METHODS: Eighty specimens, 20 for each alginate tested, were according to the manufacturers' instructions. A cutting die described in American Standard Test and Material document D-1004-94a was used to prepare test specimens. Immediately after removal from the cutting die, test specimens were measured at five points with an electronic caliper. Test specimens were placed into a specialized jig, and a Shimpo force gauge was used to measure tear strength. The tear test was performed no more than 10 minutes after mixing the alginate material with water. The tear strength was then calculated. One-way ANOVA was used to compare the mean force required to induce tearing for each of the four groups, followed by the Newman-Keuls pairwise multiple comparisons test. RESULTS: Results were considered statistically significant at p < .05. There was a statistically significant difference between alginate materials (p < .0001). The Newman-Keuls multiple comparisons procedure showed that Tare-Free Alg (514.5 g/cm) had a significantly higher tear strength value (p < .0001) than Jeltrate (259.0 g/cm), Identic (289.9 g/cm), and Kromopan (323.9 g/cm). CONCLUSIONS: Tare-Free Alg had the highest tear strength value, followed by Jeltrate, Identic, and Kromopan. PMID- 9743666 TI - An alternative to splinting multiple implants: use of the ITI system. AB - The restoration of patients with dental implants presents many challenges. Some of the common concerns and problems are difficulty in achieving a passive-fitting cast framework and loosening of prosthesis-retraining screws or abutment screws when using external hex prosthesis connection systems. The ITI system uses a unique connection of the abutment to the prosthesis-anchoring component. Because of this unique connection, many problems encountered with external hex designs have been greatly reduced or eliminated by the ability to more easily restore individual teeth with individual implants. This article will present the rationale for restoration of a patient with implants using individual implant supported restorations. PMID- 9743667 TI - Retrievable cemented implant restorations. AB - Prostheses can be attached to implants or implant abutments by screw retention, cementation, or with the use of set-screws. The indications for non-screw retained restorations are discussed with respect to implant body position and surgical limitations for implant placement. The advantages and disadvantages of each method are outlined. A technique that uses a screw threaded into the implant restoration to displace the cemented restoration is presented. Use of this technique will allow predictable removal of the cemented restorations without damage to the prosthesis or abutment. PMID- 9743668 TI - Wiropress SL: a multipurpose dental pressure vessel with transparent chamber. AB - Pneumatic vessels used in dentistry provide a pressurized environment to enhance the outcome of many laboratory procedures. A pressurized environment directs volume loss associated with polymerization shrinkage, minimizes air inclusions in powder/liquid mixtures, raises the boiling point of liquids, and facilitates flow of impression and replication materials, improving surface detail. The following report illustrates the usefulness of a reliable pneumatic vessel during denture base repair resin processing and replication of pattern details. PMID- 9743669 TI - The dental management of patients with tardive dyskinesia. AB - Tardive dyskinesia is a syndrome of involuntary muscular movements seen in patients taking antipsychotic drugs. The dental practitioner may be the first health care worker to notice the signs and symptoms. This article describes the presentation and dental management using a clinical example. PMID- 9743670 TI - The origin and evolution of Dublin's early Georgian voluntary hospitals. PMID- 9743671 TI - The simulation of clinical corrosion of endodontic files. PMID- 9743672 TI - The effect of isopropyl alcohol desiccation on apical sealability of two commonly used endodontic cements. AB - The present study was undertaken to determine the ability of alcohol desiccation to improve the apical seal by removing residual moisture from the canal. From the results of this study, isopropyl alcohol did not improve the sealability of either Roth's 801 sealer or AH26 sealer. Thus, the employment of this technique should be questioned on the basis of the potential source of irritation of the alcohol and the addition of an extraneous procedural step. PMID- 9743673 TI - Direction and magnitude of postsurgical changes following mandibular advancement with rigid fixation. PMID- 9743675 TI - The effect of elevated calcium on normal osteoblasts. PMID- 9743674 TI - Characterization and comparison of osteoblasts cultured from human bone from facial bones and the head of the femur. PMID- 9743676 TI - The fate of the odontoblasts following extirpation of the pulp from human teeth. PMID- 9743677 TI - Immunolocalization of phenotypic cartilage macromolecules within fibroblastic cellular nodules induced by chondrogenic inducing agents in vitro. PMID- 9743678 TI - Trans-surgical restoration of extensive Class IV defects in the anterior dentition. AB - Direct bonding is the most commonly utilized treatment for conservative aesthetic restoration of the anterior dentition. Class IV defects require special attention due to their high incidence, particularly in young patients. One of the major challenges for the clinician in treating defects in this category is selecting the appropriate restorative composite resin material that emulates the physical and optical characteristics of dentin and enamel and conceal the fracture line at the tooth/composite interface. Class IV defects with margins violating the biologic width present another concern, since this violation often impedes the attainment of the correct anatomic contours. The learning objective of this article is to describe a technique for achieving a correct integration of the periodontium, the tooth structure, and the restorative composite resin material to achieve a high level of aesthetic excellence. PMID- 9743679 TI - The aesthetic challenges of single tooth replacement: a comparison of treatment alternatives. AB - Several options are currently available to address the challenge of restoring a single tooth in the maxillary anterior region. To select the most appropriate treatment option for each patient, every case should be evaluated individually and all available options reviewed. The learning objective of this article is to review the treatment options and the selection of appropriate treatment modalities. The soft tissue contour around pontics and implants is discussed for efficiency, predictability, and aesthetic excellence. Three clinical cases illustrate and document three different treatment modalities in the maxillary anterior region--a single tooth implant-supported restoration, combined with porcelain laminate veneers; an adhesive bridge, combined with a porcelain laminate veneer; and a conventional porcelain-fused-to-metal bridge. PMID- 9743680 TI - Prognosis of extreme skeletal discrepancies: a case report. PMID- 9743681 TI - Single implant restorations: prosthetically induced soft tissue topography. AB - An aesthetic transition from the smaller diameter of the implant to the prosthetic restoration that resembles the size of the natural tooth has presented an ongoing challenge to the implant restorative dentists. The appearance of the surrounding soft tissue is of major importance, and various techniques have been developed to guide and optimize its topography. The learning objective of this article is to present a cervical contouring concept, whereby the soft tissue topography is optimally determined already in the laboratory phase. Using a custom abutment and provisional crown as components of the transmucosal prosthetic unit (TPU), the topography is transferred to the vital intraoral tissues, which predictably adapt to the enhanced aesthetic configuration. Clinical cases are presented to demonstrate the sequence of the technique in treating the anterior region of the maxilla. PMID- 9743682 TI - Rest in peace, G.V. Black: Part I. PMID- 9743683 TI - Reattachment of coronal fragments: operative considerations for the repair of anterior teeth. AB - The maxillary incisors occupy an extraordinary position in the dental arch. They set the aesthetic tone, and their influence on the overall well-being of the individual cannot be overemphasized. However, their eruptive pattern and dominance carries a significant risk for trauma, particularly in childhood. More than half of all traumatic dental injuries involve the central incisors. In the past, fractured teeth were either extracted, trimmed and leveled, or restored with cast restorations. The learning objective of this article is to stimulate a shift in treatment strategy towards aesthetic reattachment of fractured segments. Indications and limitations for reattachment are outlined, including the primary and secondary restorative efforts. Innovative operative techniques that improve the aesthetic, biologic, and mechanical variables of the reattachment treatment option are presented. PMID- 9743684 TI - First call your colleague. PMID- 9743685 TI - Indirect composite restorations in the anterior region: a predictable technique for complex cases. AB - The indirect composite inlay technique demonstrates excellent aesthetic results in the posterior regions, achieved by reproducing the shape, color, and contour of the natural tooth on a cast model rather than using the free-hand composite restorative modality. The utilization of the indirect technique can be adapted for the anterior regions as well, and it offers a valid treatment alternative that is predictable with optimal restorative results. The evaluation, planning, and accuracy of implementation of the various phases of this methodology allow a more precise achievement of the result; any potential error in the restoration can be corrected prior to final cementation. This technique increases the quality of composite restorations that have always been considered unpredictable and extensively dependent upon the skills of the operator. The learning objective of this article is to familiarize the reader with the clinical and laboratory phases of this indirect treatment modality of the anterior dentition. PMID- 9743686 TI - Excellence with simplicity in aesthetic dentistry. PMID- 9743687 TI - Transdermal drug application. PMID- 9743688 TI - Aesthetic considerations in endodontics: internal bleaching. AB - During endodontic treatment of any tooth, aesthetics must be considered in the same manner as during any other dental treatment. The most common aesthetic challenge associated with endodontics is the discoloration of natural tooth structure. The discoloration may be a result of pulp pathosis, especially pulpal hemorrhage prior to or during treatment, or it may be due to various endodontic and restorative materials placed in the pulp chamber. There are several simple measures that can be utilized during and following endodontic treatment to eliminate or reduce aesthetic deficiency. The learning objective of this article is to discuss internal bleaching of discolored pulpless teeth that have been endodontically treated. The discussion includes the chemical composition of bleaching agents and principles by which they function during the bleaching procedures. PMID- 9743689 TI - ADA Council on insurance white paper on Medical Savings Accounts. PMID- 9743690 TI - Svirsky on infection control. PMID- 9743691 TI - Sexual dysfunction associated with the management of prostate cancer. AB - Improvements in the management of prostate cancer have increased the need to consider patients' quality of life, in particular in relation to sexual function. The causes of sexual dysfunction are varied and derive from both the condition and its management. Health professionals must choose their treatment strategies with great care. Patient expectations must be understood, and patients should be offered counselling, as an understanding of what can reasonably be expected contributes to patients' perception of their quality of life. There are few studies on sexual dysfunction in patients with prostate cancer. A first step would be to develop reliable questionnaires for the assessment of the problem. This article describes and discusses the findings of one such recently developed questionnaire. When baseline measures of sexual function have been established and the extent of sexual dysfunction in patients with prostate cancer is reliably quantified, large multicentre trials can be performed to evaluate the impact of different therapies on sexual function and quality of life. Sexual dysfunction is an area which will be of increasing importance to urologists who manage prostate cancer and one that should not be underestimated. PMID- 9743692 TI - Improvement of medical care quality after implementation of a clinical path monitoring program for transurethral prostatectomy patients. AB - OBJECTIVES: The purpose of this study was to determine the effect on quality of care through the implementation of a clinical path for patients receiving transurethral prostatectomy. METHODS: We selected ten quality indicators with important clinical relevance as representative elements of the clinical path. These quality indicators were monitored during the entire hospitalization period of 100 consecutive patients who received transurethral prostatectomy. Monitoring data obtained from these patients were compared to data from 100 patients who received transurethral prostatectomy prior to implementation of the clinical path. Data was assessed to determine the relationship between quality indicators and management processes. RESULTS: Implementation of the clinical path for transurethral prostatectomy significantly decreased the percent of patients with incomplete preoperative tests on admission day, the duration of intravenous antibiotics administration, the percent of patients who required acute pain management postoperatively, the percent of patients who received postoperative bladder irrigation with normal saline and the percent of patients who had their Foley catheter removed after postoperative day 2. Three of the quality indicators had a significant relationship with management processes and may have directly affected the total admission charges. CONCLUSIONS: To evaluate the effect of the transurethral prostatectomy clinical path implementation on the quality of medical care, we compared ten quality indicators before and after implementation of this path. We concluded that implementation of the clinical path resulted in a statistically significant improvement in the quality of medical care. PMID- 9743694 TI - Increase of renal cell carcinoma incidence in central Europe. AB - OBJECTIVE: In recent years the incidence of renal cell carcinoma (RCC) diagnosis has increased about 15-20%. It remains to be established whether this increase of incidence is reality or not. The main aim of this study was to analyze the reason for the increase of incidence. METHODS: In the present study, 23, 247 autopsies performed in the years 1985-1995 in the area of Jena (Germany) (14,793 autopsies) and Hradec Kralove (Czech Republic) (8,454 autopsies) were analyzed. RESULTS: In this autopsy series comprising 23,247 autopsies, the percentage of patients who died of RCC is 1.76% in Jena and 1.55% in Hradec Kralove (200,000 inhabitants each). Over this time the incidence of RCC in autopsies has increased. CONCLUSION: In spite of the increased amount of incidentally found RCCs since beginning widespread use of ultrasonography, the percentage of clinically recognized RCCs in the total of all found RCCs in autopsies is nearly constant over the 11-year period in Jena and 10-year period in Hradec Kralove. Thus, the increased number of radical nephrectomies is not only caused by widespread use of ultrasonography. The increasing trend of the incidence of RCC seems to be real. PMID- 9743693 TI - Cost-effective emergency diagnosis plan for urinary stone patients presenting with ureteric colic. AB - OBJECTIVE: To develop a cost-effective plan for the accurate diagnosis of urinary stone patients presenting with ureteric colic based on an assortment of investigations which are less invasive and more economical than intravenous urography (IVU). PATIENTS AND METHODS: 143 consecutive emergency patients presenting with ureteric colic were admitted to hospital and prospectively studied by history recording, physical examination, laboratory tests and imaging procedures according to a preset format. Significant association of the final diagnosis of urinary stones (which was made by actual stone retrieval) with various diagnosis variables obtained from the results of investigation (including IVU) was statistically studied using bivariate correlation and multivariate logistic regression analysis. Algorithms for reaching an accurate diagnosis of urinary tract stones were formulated using the most significant diagnostic variables and the accuracy of each of those plans was compared with that of emergency IVU. RESULTS: 18 patients were excluded for various reasons. Of the remaining 125 patients 82 (66%) were confirmed as having urinary stones. A positive IVU had the strongest correlation with the final diagnosis of urinary tract stones. Other findings associated with eventual stone retrieval in a descending order of significance were: calcular sonographic features; radio opacities on a plain abdominal film of the kidney, ureter and bladder (KUB), and microhaematuria. Based on these findings two algorithms could be formulated to reach as accurate a diagnosis as possible. Algorithm A in which an initial ultrasound is mandatory had a sensitivity of 89%, a specificity of 88% and an overall accuracy of 88% for urinary stone detection compared with 91, 77, and 86%, respectively, for algorithm B in which ultrasonography was employed selectively after initial KUB and urinalysis for microhaematuria. This compares with 94, 79, and 89%, respectively, for IVU. CONCLUSION: Both plans are viable alternatives which could replace routine emergency IVU. PMID- 9743695 TI - Impact of transition zone biopsies in detection and evaluation of prostate cancer. AB - OBJECTIVE: To analyze the impact of 2 systematic transition zone (TZ) biopsies in addition to systematic sextant biopsies in an effort to establish the importance of cancer detected in the transition zone. METHODS: Between November 1995 and October 1996, TRUS-guided systematic sextant peripheral zone (PZ) and two additional TZ biopsies were performed on 189 consecutive men. Radical retropubic prostatectomy (RRP) was performed to 13 patients with organ-confined prostate cancer. The biopsy results of the 52 patients with cancer and the pathological specimens of the patients who underwent surgery were compared. RESULTS: Of the 189 patients, 52 (27.5%) had prostate cancer of whom 20 (38.5%) both in the PZ and TZ, 31 (59.6%) only in the PZ, and 1 (1.9%) in the TZ only. Of the 96 patients with high serum PSA levels despite normal DRE, 14 had prostate cancer. TZ cancer only rate was 7.1% (1 in 14 patients) in this group. RRP was performed to 8 patients who had cancer only in the PZ and 5 patients in both TZ and PZ. The pathological stages of the postoperative specimens and extracapsular extension rates of those with cancer in the PZ and TZ were significantly higher (p = 0.029 and p = 0.008, respectively). CONCLUSIONS: Routine TZ biopsy does not substantially increase the prostate cancer detection rate, however it can be useful in selected patient groups. If further studies reveal the relationship of cancer in the transition zone, higher capsular extension rate (pT3 cancer) and higher pathological stage after radical surgery, then TZ biopsies may yield additional information that might influence the therapeutic approach. PMID- 9743696 TI - Enhanced expression of prostate-specific antigen in the transition zone of the prostate. A characterization following prostatectomy for benign hyperplasia. AB - OBJECTIVE: To determine whether the serum levels of total prostate-specific antigen (t-PSA), free PSA (f-PSA) and PSA complexed to alpha 1-antichymotrypsin (PSA-ACT) result from different expressions in various prostatic zones. METHODS: In a series of 127 consecutive men undergoing transurethral resection of the prostate (TURP) for BPH between May 1995 and February 1996, t-PSA, f-PSA (ProStatus, Wallac) and PSA-ACT were measured before and 3-4 months after surgery. Pre- and postoperative prostate volumes were measured by TRUS. Resected tissue was assumed to be the transition zone (TZ) while postoperative volume was defined as peripheral zone (including the central one) (CPZ). Pre- and postoperative serum PSA was related to pre- and postoperative volume and resected tissue to the difference between pre- and postoperative serum PSA, respectively. The serum PSA per 1 g tissue was calculated. Group I consisted of 96 historically proven BPH with no signs of inflammation, group II of 19 BPH patients with transurethral catheters inserted sometime prior to surgery to relieve urinary retention, and group III of 12 patients with incidental carcinomas. RESULTS: In patients undergoing TURP without prior catheterization (group I) t-PSA (group I) declined from median 3.43 to 0.96 ng/ml after TURP by 72%, even though the prostate volume did so only by 44%, whereas the ratio free-to-total (f/t) PSA remained stable (median 24.9% pre- vs. 26.6% postoperatively). The TZ expressed approximately 2.7-fold more t-PSA than the remaining CPZ: median 0.14 vs. 0.052 ng/ml/g, respectively, and as to f-PSA it did so likewise: median 0.032 vs. 0.012 ng/ml/g, respectively. With transurethral catheterization prior to surgery (group II) the t-PSA density within whole prostate increased 1.4-fold as compared to this density without such catheterization: from median 0.089 (group I) to 0.13 ng/ml/g tissue, respectively (p < 0.007), and within the TZ alone 1.6-fold elevation from median 0.14 to 0.23 ng/ml/g, respectively (p < 0.02) was observed. In incidental carcinoma (group III) a reduced ratio of f/t PSA of 11.7% in the TZ as compared to 22.1% in the CPZ (22.1%) was observed. CONCLUSIONS: In BPH both t PSA and f-PSA are predominantly expressed within the TZ, which could help to improve the specificity of the PSA density in cancer detection by using the sum of the t-PSA densities of the TZ and CPZ: (0.14 ng/ml/g x TZ) + (0.052 ng/ml/g x CPZ). It is the first time that the supposed origin of the incidental carcinoma (from the TZ) is confirmed biochemically by a f/t PSA ratio exclusively reduced in the TZ but not in the CPZ. The post-TURP unchanged free-to-total ratio (26.6%) may be useful for the early detection of cancer in patients followed up after TURP. PMID- 9743697 TI - A comparative study of a double-line versus a fan-shaped technique for obtaining transrectal ultrasound-guided biopsies of the prostate. AB - INTRODUCTION AND OBJECTIVES: Transrectal ultrasound-guided biopsy of the prostate is an established method to obtain prostate specimens for histological analysis. The aim of this prospective study was to compare the prostate cancer detection rate of the conventional double-line biopsy to a more fan-shaped biopsy technique. METHODS: A total of 107 men were included in this study. The indication for performing a prostate biopsy was a serum prostate-specific antigen level exceeding 4 ng/ml and/or suspicious findings on digital rectal examination. 53 patients were biopsied by the conventional double-line technique (method A): 3 biopsies in the midparasagittal plane from each lobe at the apex, middle and basis, at an angle of approximately 45 degrees C. 54 patients were biopsied with the so-called fan-shaped technique (method B): 6 biopsies were taken from the left to the right lateral margin in one plane at the same angle. RESULTS: 642 prostate biopsy cores were obtained, a subset of 133 biopsies were identified to yield prostate cancer. The percentage of positive biopsies was higher in group B (n = 81; 61%) as compared to group A (n = 52; 39%). The overall prostate cancer detection rate by the fan-shaped technique was 37% (20/54) as compared 30.1% (16/53) to the conventional double-line technique, but this difference did not reach statistical significance (p < 0.05) presumably because of the small number of patients. CONCLUSION: These data suggest that the fan-shaped biopsy technique seems to have a higher cancer detection rate and a higher number of positive core biopsies than the conventional double-line technique, because of more presence in the apex and the peripheral zone of the prostate where most prostate cancers originate from. PMID- 9743698 TI - Prognostic factors in clinical stage I nonseminomatous germ cell testicular tumors: rationale for different risk-adapted treatment. AB - OBJECTIVE: Surveillance after orchiectomy alone becomes popular for the management of clinical stage I nonseminomatous germ cell testicular tumours (CS I NSGCTT). Effort to identify patients at high risk of relapse leads to searching prognostic factors of CS I NSGCTT. The aim of this study was to identify those patients in whom a surveillance policy is less likely to be successful. PATIENTS AND RESULTS: Seventy-two CS I NSGCTT patients were stratified to different risk adapted therapeutic approaches according to histopathologic findings of primary tumor removed by inguinal orchiectomy. Eighteen patients (group A) with vascular invasion and majority of embryonal carcinoma component in the primary tumor were treated with adjuvant BEP chemotherapy. None of them experienced disease progression after a median follow-up period of 36 months after orchiectomy. Five patients (group B) with vascular invasion and the majority of teratomatous elements in the primary tumor have been followed up 56 months after orchiectomy. They were treated with primary retroperitoneal lymph node dissection (RPLND). Two of them (40%) had pathologic stage II after RPLND and underwent subsequent chemotherapy. One of them died due to disease progression 29 months following orchiectomy. Another one lives with no evidence of disease (NED). Three patients in pathologic stage I are alive with NED. Forth-nine patients (group C) without vascular invasion have been followed up for a median duration of 37 months after orchiectomy. They were kept under close surveillance, consisted of regular follow up with tumor markers, chest x-ray and CT of the retroperitoneum. Disease progression was observed in 7 (14.3%) patients after a median duration of 8 months after orchiectomy. They were treated with BEP chemotherapy and live with disease-free median survival of 22 months after completion of therapy. The overall survival rate of all 72 patients was 98.6%. The median survival for all patients was 37 months (range 7-73). CONCLUSIONS: The authors will continue to use surveillance policy only in patients without vascular invasion in the primary tumor. PMID- 9743699 TI - Incidence and characteristics of antiandrogen withdrawal syndrome in prostate cancer after treatment with chlormadinone acetate. AB - OBJECTIVES: In patients with progressive prostate cancer who have been treated with surgical or medical castration plus an antiandrogen, antiandrogen withdrawal can result in a significant decline in serum prostate-specific antigen (PSA). Although the incidence of antiandrogen withdrawal syndrome after combination treatment with the nonsteroidal antiandrogen flutamide has been thoroughly documented, the phenomenon clearly occurs in many other combination therapies and is presently being widely investigated. This paper would like to contribute to this effort by describing the endocrine withdrawal phenomenon in patients treated with combinations of castration plus chlormadinone acetate, ethynylestradiol or estramustine phosphate. MATERIALS AND METHODS: Clinical records of 68 prostate cancer patients who had been treated with surgical castration plus the administration of chlormadinone acetate, ethynylestradiol or estramustine phosphate, and who had shown clinical progression associated with a steady increase in serum PSA, were investigated. Forty-one cases were evaluable for changes in PSA after discontinuation of the hormonal agents. RESULTS: Of 28 patients who had been treated with chlormadinone acetate, 12 (42.9%) revealed 50% or more decline in PSA level following withdrawal of the agent. Among these, 5 cases (17.9%) showed subjective and/or objective improvements. There was no significant difference in histological grade of the tumor at diagnosis, mode of progression, time interval from the start of endocrine therapy to discontinuation of the hormonal agents, or PSA level at withdrawal of the agents between patients who did develop antiandrogen withdrawal syndrome and those who did not. CONCLUSION: When a steady increase in serum PSA is noted in a prostate cancer patient who has been treated with castration plus a steroidal antiandrogen, discontinuation of the antiandrogen may benefit the patient. PMID- 9743700 TI - Management of renal angiomyolipoma: analysis of 15 cases. AB - OBJECTIVE: We present our experience with 15 patients with renal angiomyolipoma warranting intervention. METHODS: The medical records and radiological studies were reviewed for patient age and sex, tumor location and size, association with tuberous sclerosis, and treatment approach. All patients were regularly followed by ultrasound and computed tomography scan. RESULTS: Presenting symptoms were retroperitoneal bleeding in 9 patients and flank pain in 6. Excluding cases of tuberous sclerosis (mean tumor diameter 11 cm), the mean diameter of the two tumors that bled was 5.4 cm, similar to those in the patients presenting with flank pain. Two patients with retroperitoneal bleeding had tumors < 3 cm. Angioinfarction was performed in 7 patients, partial nephrectomy in 3, and total nephrectomy in 4. One patient with tuberous sclerosis, who was observed only, died of bleeding and sepsis. The mean follow-up period of 4.3 years revealed stable creatinine levels and no recurrent hemorrhage. CONCLUSIONS: The management approach of angiomyolipoma should be aimed at parenchymal preservation which can be effectively accomplished by limited surgery or preferably by selective embolization. Preventive embolization may be feasible even for small tumors. However, any doubt about the diagnosis of angiomyolipoma should be clarified by surgery. PMID- 9743701 TI - Hemodynamic monitoring of the contralateral testis during unilateral testicular torsion describes the mechanism of damage. AB - Contralateral testicular perfusion during unilateral testicular torsion was evaluated using simultaneous blood flow and O2 content determinations. Two groups, each consisting of 7 rats, were studied. Sham operation or 720 degrees clockwise twisting was performed on the left testes, and blood flow, O2 content and temperatures were monitored in the right testes for 180 min. An ultrasonic perivascular Doppler flowmeter system, an electronic thermometer and an O2 electrode were used for the monitoring. The contralateral testicular blood flow and relative O2 contents were stable in the control group. However, the initial and 180 min blood flow values decreased from 0.21 +/- 0.04 to 0.11 +/- 0.02 ml/min (p < 0.001), and the O2 contents from 0.857 +/- 0.123 to 0.319 +/- 0.037 (1.0 corresponds to 19.6 mm Hg pO2, p < 0.05) in the experimental group. Unilateral testicular torsion decreases the blood flow and O2 content of the contralateral testis. The contralateral testicular injury encountered following unilateral testicular torsion might result from hypoxia following the decrease in blood flow. PMID- 9743702 TI - Residues of some veterinary drugs in animals and foods. Monographs prepared by the forty-eighth meeting of the Joint FAO/WHO Expert Committee on Food Additives. PMID- 9743703 TI - Carbohydrates in human nutrition. Report of a Joint FAO/WHO Expert Consultation. PMID- 9743704 TI - Validation of analytical methods for food control. Report of a Joint FAO/IAEA Expert Consultation. PMID- 9743705 TI - Compendium of food additive specifications. Addendum 4. Joint FAO/WHO Expert Committee on Food Additives 46th session. PMID- 9743706 TI - Compendium of food additive specifications. Addendum 5. Joint FAO/WHO Expert Committee on Food Additives 49th session. PMID- 9743707 TI - Antimicrobial use principles drafted; steering committee invites comments. PMID- 9743708 TI - Draft AVMA principles on judicious therapeutic use of antimicrobials. American Veterinary Medical Association. PMID- 9743709 TI - Aminoglycosides: delegates show resolve. PMID- 9743710 TI - Vaccination guidelines referred back. PMID- 9743711 TI - Fluoroquinolone approved for cattle. PMID- 9743712 TI - Symposium devoted to vaccine-associated feline sarcomas. PMID- 9743713 TI - Debate regarding vaccination procedures and homeopathy. PMID- 9743714 TI - Medical treatment versus surgery for hiatal hernias. PMID- 9743716 TI - The use of persuasion by food animal veterinarians. PMID- 9743715 TI - Warning against use of some permethrin products in cats. PMID- 9743717 TI - What is your diagnosis? Primary or metastatic neoplasia, or bacterial or fungal osteomyelitis. PMID- 9743718 TI - Theriogenology question of the month. Benign prostatic hypertrophy (BPH), prostatitis, and prostatic neoplasia. PMID- 9743719 TI - Economic evaluation of neonatal health protection programs for cattle. AB - OBJECTIVE: To develop an economic tool that can be used to help cattle producers evaluate benefits of neonatal health programs. DESIGN: Computer simulation of a multiple-year spreadsheet model, using economic and production variables. SAMPLE POPULATION: Records for a university research farm beef herd. PROCEDURE: Data from the university research farm beef herd for each year from 1990 to 1995 were evaluated to determine economic benefits for the cow-calf enterprise that would result from a decrease in morbidity and mortality. A baseline economic evaluation of returns to variable costs was performed, using actual production and marketing information. Actual economic performance was contrasted with a projected simulation in which morbidity and mortality were decreased. Sensitivity analysis for the simulation model assessment of a neonatal health program was also performed. RESULTS: Mean-per-cow increase in net income for the herd during the 6 year period for morbidity and mortality reductions of 20, 40, and 60% was $7.44, $14.93, and $22.42, respectively. Sensitivity analysis revealed that net income per cow was not sensitive to errors in projections of morbidity and mortality. CLINICAL IMPLICATIONS: Identifying potential economic benefits for implementing a neonatal health plan and quantifying the costs to implement each component of the plan can be used by veterinarians and their clients when formulating a proactive strategy to provide the greatest potential for economic reward. PMID- 9743721 TI - Gastrin concentrations in plasma of cats with chronic renal failure. AB - OBJECTIVE: To determine the prevalence of hypergastrinemia in cats with naturally developing chronic renal failure (CRF) and the correlation between gastrin concentration in plasma and severity of CRF. DESIGN: Cohort study. ANIMALS: 30 cats with naturally developing CRF and 12 clinically normal control cats. PROCEDURE: Gastrin concentrations in plasma were determined by double-antibody radioimmunoassay of blood samples obtained from cats after food was withheld 8 hours. Concentrations were compared, using a nonparametric Kruskal-Wallis ANOVA. RESULTS: 18 cats with CRF had high gastrin concentrations (median, 45 pg/ml; range, < 18 to > 1,333 pg/ml), compared with those for control cats (< 18 pg/ml). Prevalence of hypergastrinemia increased with severity of renal insufficiency. Three of 9 cats with mild CRF, 6 of 11 cats with moderate CRF, and 9 of 10 cats with severe CRF had high gastrin concentrations. Gastrin concentrations were significantly different between control cats and cats with CRF, regardless of disease severity. CLINICAL IMPLICATIONS: The potential role of high concentrations of gastrin on gastric hyperacidity, uremic gastritis, bleeding from the gastrointestinal tract, and associated clinical signs of hypergastrinemia (e.g., anorexia and vomiting) may justify use of histamine2 receptor antagonists or proton pump inhibitors to suppress gastric acid secretion in cats with CRF that have these clinical signs. PMID- 9743720 TI - Systemic hypertension and proteinuria in dogs with diabetes mellitus. AB - OBJECTIVE: To determine prevalence and severity of systemic arterial hypertension and proteinuria in dogs with naturally developing diabetes mellitus (DM) and to determine whether these abnormalities were related to age, sex, duration of DM, or degree of control of glycemia. DESIGN: Case series and cohort study. ANIMALS: Fifty dogs with naturally developing DM. PROCEDURES: Blood pressure was measured in all 50 dogs. Thirty-eight dogs were evaluated once, and 12 were evaluated sequentially. Thirty-five were evaluated for proteinuria by determining protein to-creatinine ratio in urine (n = 35) or by electrophoresis of urine (33). RESULTS: Hypertension was detected in 23 on the basis of a systolic pressure > 160 mm HG (12 dogs), a diastolic pressure > 100 mm HG (21), or a mean pressure > 120 mm HG (23). All dogs with systolic hypertension had concurrent diastolic and mean hypertension, and 19 of 21 dogs with diastolic hypertension had concurrent high mean pressure. Ten of 12 dogs reevaluated at subsequent visits had no change in blood pressure. Blood pressure remained consistent in 3 dogs tested at different times during the day on a single visit. Duration of DM and presence of proteinuria were significant predictors of hypertension. Seven of 35 (20%) dogs had an increased protein-to-creatinine ratio in their urine. Albumin concentration and albumin-to-creatinine ratio were significantly higher in urine from diabetic dogs, compared with healthy, nondiabetic dogs. Hypertension was associated with an increased albumin-to-creatinine ratio. CLINICAL IMPLICATIONS: Systemic hypertension and proteinuria may be common in diabetic dogs, but the clinical importance of these findings are, as yet, unknown. PMID- 9743722 TI - Aortic foreign body resulting in ischemic neuromyopathy and development of collateral circulation in a cat. AB - A cat evaluated for paraplegia had firm pelvic limb musculature and did not have femoral pulses. External wounds were not evident, but abdominal radiography revealed a round metallic foreign body on the midline ventral to the sixth lumbar vertebra. Angiography indicated stenosis or thrombosis of the aorta in association with the foreign body; collateral circulation arose from the fifth lumbar artery. Arteriotomy was performed to extract the foreign body and associated thrombi. Six weeks after surgery, angiography revealed blood flow in the abdominal portion of the aorta, but no evidence of obstruction or additional collateral vessels. The cat regained function of the pelvic limbs within 1 year after surgery. Ischemic neuromyopathy and paraplegia in cats is commonly associated with aortic thromboembolism. A thrombus is necessary to cause typical clinical signs, and vasoactive substances released by platelets in the thrombus are believed to cause ischemic neuromyopathy. Progression of the collateral circulation may allow for clinical improvement without surgical intervention. PMID- 9743723 TI - Surgical treatment of urate calculi in Dalmatians: 38 cases (1980-1995). AB - OBJECTIVE: To assess clinical signs and response to surgical treatment in Dalmatians with urate urolithiasis. DESIGN: Retrospective study. ANIMALS: 38 Dalmatians. PROCEDURE: Medical records from 1980 to 1995 of Dalmatians with urate urolithiasis were reviewed to obtain information on history, results of physical examination, hemogram, biochemical analysis, urinalysis, bacterial culture of urine, diagnostic imaging, analysis of calculi, treatment, and recurrence. RESULTS: 35 (92%) dogs were males. Mean age at admission was 4.9 years. Common clinical findings and initial complaints included dribbling of urine, stranguria, vomiting, tense abdomen with signs of pain, and a large bladder. Hematuria was found in 85% of dogs in which urinalysis was performed. Crystalluria was found in 54% of dogs. Bacteria were isolated from urine from 36% of dogs. Contrast radiography and abdominal ultrasonography were the most sensitive diagnostic tests for uroliths. Dogs that underwent scrotal urethrostomy and cystotomy had the fewest number of recurrent clinical signs that were attributable to urinary calculi. Clinical recurrence rate in dogs on a protein-restricted diet was 27%, compared with that (36%) for dogs on a commercial diet. CLINICAL IMPLICATIONS: Urate urolithiasis is more commonly recognized in male Dalmatians compared with females. Contrast radiography and ultrasonography appear to be the most useful techniques for detecting urate uroliths. Scrotal urethrostomy and cystotomy was the most effective surgical treatment for preventing recurrence of clinical signs associated with calculi. Complete removal of calculi and protein-restricted diets may have a beneficial effect in reducing recurrence of calculi. PMID- 9743724 TI - Passive transfer, rate of decay, and protein specificity of antibodies against equine arteritis virus in horses from a Standardbred herd with high seroprevalence. AB - OBJECTIVE: To determine rate of decay of passively acquired antibodies in Standardbred foals on a farm with a high seroprevalence to equine arteritis virus (EAV) and to determine whether vertical or horizontal transmission of the virus was responsible for infection on the farm. DESIGN: Repeated-measures study. ANIMALS: 46 Standardbred horses (15 brood mares and their foals, 5 stallions, and 11 young horses). PROCEDURE: Serum samples obtained from horses on the farm were evaluated by serum neutralization and western immunoblot analysis to detect EAV specific antibodies. The half-life of passively acquired antibodies in foals was estimated by use of regression analysis. RESULTS: Most (14/15) of the mares evaluated were seropositive to EAV. After suckling, their foals were also seropositive. Mean biological half-life for passively acquired antibodies in serum samples obtained from foals was 32 days (r2 = 0.61). The foal born to a seronegative dam and all 11 young horses from the farm were seronegative to EAV. At least 2 of 5 stallions on the farm were persistently infected carriers that were shedding virus in their semen. Immunoblot analysis of seropositive serum samples most consistently recognized the M protein of EAV. CLINICAL IMPLICATIONS: Analysis of these data indicated that a modified-live EAV vaccine can be administered to foals after they are 8 months old without risk of interference from maternal antibodies, regardless of serologic status of the foal's dam. Horizontal transmission of EAV via the respiratory tract apparently was uncommon on the farm, indicating that mares primarily were infected by venereal transmission of virus from carrier stallions. PMID- 9743725 TI - Medical management of urinary calculi in a stallion with breeding dysfunction. AB - A 9-year-old Thoroughbred stallion was examined because of breeding dysfunction and possible urethritis. The stallion had good libido and readily obtained an erection, mounted, and intromitted but did not thrust and ejaculate. After mounting the mare, the stallion would squeal and dismount. Endoscopic examination of the urethra and bladder revealed irregular, spiculate yellow crystals (< 1 cm in size) and sabulous deposits; numerous calculi were embedded in the mucosa of the bladder. Because the horse was at the start of a breeding season, the owner would not give permission for general anesthesia. Medical management was attempted, because postoperative convalescence after surgical removal of calculi might have curtailed breeding activities, and the calculi were small. Every 1 to 3 days, the bladder was lavaged with saline solution containing acetic acid, and anti-inflammatory and antimicrobial drugs were administered. The stallion was able to return to breeding mares, and sperm numbers and semen quality were good. However, urine contamination of the ejaculate was detected, suggesting that the stallion may have had a primary neurologic deficit affecting bladder control and function that was causing calculi to form secondarily because of delay in movement of urine through the urinary tract. PMID- 9743726 TI - Complete fractures of the third metacarpal or metatarsal bone in horses: 25 cases (1980-1996). AB - OBJECTIVES: To compare treatments of complete fractures of the third metacarpal (MC) or metatarsal (MT) bone in horses and to identify factors that could impact prognosis. DESIGN: Retrospective case series. ANIMALS: 25 horses with fractures of the third MC or MT bone that were treated by use of internal fixation, external coaptation, or both. PROCEDURE: Medical records from the Veterinary Medical Data Base of horses treated for fractures of third MC or MT bone at Texas A&M University from 1980 to 1994 and Purdue University from 1980 to 1996 were reviewed. Information on signalment, results of physical and radiographic examinations, treatment, and outcome were obtained. For horses that had radiographic evidence of healing, long-term follow-up information was obtained by telephone contact with owners or referring veterinarians. RESULTS: Age, sex, weight, and limb affected were not related to outcome; however, affected horses were younger than the general hospital populations. Seventeen horses had open fractures at referral. Infection was the most common complication after surgery, with open fractures more likely to become infected. Nonunion in an infected fracture was the most common reason for postoperative failure (7 horses). Long term follow-up was available for 16 horses; 11 of these had no complications related to surgical repair. CLINICAL IMPLICATIONS: Fractures of the MC or MT bone are not always associated with a poor prognosis in horses. Proper case selection, rigid fracture stabilization, and efforts to prevent or treat infection will improve success rate. PMID- 9743727 TI - Effect of tuberculosis on milk production in dairy cows. AB - OBJECTIVE: To evaluate differences in milk production between cows with positive and negative caudal fold tuberculin test results in a Mycobacterium bovis infected dairy herd. DESIGN: Cross-sectional epidemiologic survey. ANIMALS: 369 Holstein cows with lactation duration between 200 and 360 days. PROCEDURE: The caudal fold tuberculin test was performed. Information on milk production data, parity, calving season, days of lactation, previous milk production, and whether cows had clinical mastitis was obtained from farm records. Composite milk samples were collected and submitted for bacterial culture. RESULTS: 170 cows had positive tuberculin test results, and 199 had negative results. Cows with positive test results produced less milk (mean, 347 kg [763 lb]) than did cows with negative test results after adjusting for variables biologically related to milk production. Calving season (spring, summer) was significantly associated with reduced milk production. Cows with clinical mastitis produced less milk than did cows without clinical mastitis, but the difference was not statistically significant in the multiple regression analysis. CLINICAL IMPLICATIONS: In this herd, tuberculosis was associated with a 4% decrease in milk production. Milk production losses per cow attributable to calving season (spring, summer) were 3 times those attributable to tuberculosis. However, because of the high prevalence of tuberculosis in this herd, the impact of tuberculosis on total herd milk production was an additional cause for concern. PMID- 9743728 TI - Characteristics of dairy cows during episodes of bacteriologically negative clinical mastitis or mastitis caused by Corynebacterium spp. AB - OBJECTIVES: To use clinical and lactational characteristics to determine whether bacteriologically negative (BN) clinical mastitis episodes are more apt to be caused by gram-negative or gram-positive bacteria, and to investigate severity of clinical mastitis caused by Corynebacterium spp (COR). DESIGN: Case series. SAMPLE POPULATION: 300 clinical mastitis episodes affecting 123 dairy cows vaccinated against lipopolysaccharide core antigens. PROCEDURE: Cows were examined at onset of clinical mastitis, and 23 characteristics, including rectal temperature, heart rate, rumen contraction rate, degree of dehydration, udder and milk characteristics, lactation number, stage of lactation, and season of year, were recorded. Milk production and milk constituent concentrations before onset of mastitis were obtained from herd records. Values for cows with BN milk were compared with values for cows from which milk yielded gram-negative bacteria (GNB) or gram-positive cocci (GPC); logistic regression was used to predict which pathogen type was causing BN mastitis. Characteristics for cows from which milk yielded COR were compared with those of cows from which milk was BN or yielded GPC. RESULTS: BN clinical mastitis episodes differed significantly from episodes caused by GPC, and were similar to, but milder than, episodes caused by GNB. COR were isolated in a substantial proportion of mastitis episodes, but clinical signs were milder than when GPC were isolated. CLINICAL IMPLICATIONS: Most BN mastitis episodes in cows receiving lipopolysaccharide core antigen vaccines appear to be caused by low-grade infection with GNB, and treatment and management decisions should be made accordingly. The COR may be economically important clinical mastitis pathogens in some herds. PMID- 9743729 TI - Histologic features and results of virus isolation tests of tissues obtained from teat lesions that developed in dairy cattle during winter. AB - OBJECTIVE: To determine microscopic features and involvement of viruses in teat end lesions (TEL) of dairy cows during winter. SAMPLE POPULATION: Teats with TEL on lactating Holstein cows and from udders of carcasses. PROCEDURE: Tissues obtained from TEL of 10 teats from 7 cows on 2 research farms during the winter of 1994 to 1995 and 13 teats with TEL excised from udders of carcasses at an abattoir during February 1995 were submitted for virus isolation. During the winter of 1995 to 1996, an increased prevalence of TEL was observed in a research herd. After a decrease in ambient temperature, TEL were identified, and a full thickness section of epidermis was removed from skin surrounding teat orifices. Tissues were examined by use of light and electron transmission microscopy. RESULTS: Viruses were not isolated from TEL tissues. Lesions ranged from mild elevations of the epidermis to thickened oval regions that encircled the teat orifice. The most severe lesions were dark and had thick crusts. Histologically, TEL were composed of thickened regions of epidermis most notably caused by hyperplasia of cells within the stratum spinosum. Excess production of keratinocytes was also evident, and the keratinocyte layer often contained bacteria. Ultrastructurally, squamous cells contained large amounts of keratin, but virions were not detected. Evidence of a viral etiologic agent for TEL was not detected. CLINICAL IMPLICATIONS: Development of TEL may be associated with decreases in ambient temperature. Numerous bacteria were evident in the keratin of TEL. Lesions and associated bacteria may predispose cows to mastitis. PMID- 9743730 TI - AIDS impact. PMID- 9743731 TI - HIV-related politics in long-term perspective. AB - Some long-term, large-scale socio-economic changes may affect the politics of HIV and other emerging viruses such as hepatitis C. It is useful to ask why the potential peace dividend of the early 1990s failed to provide adequate resources for HIV-related social and medical service delivery in developed or developing nations. This failure can be understood by looking at long-term global economic trends and the pressures they put on governments and corporations. They have produced a period in which fundamental issues of political and economic structure are at stake and, often, the response is a divide-and-rule politics to promote stability. National politics differ in terms of the extent to which such a 'politics of scapegoating' is institutionalized and in terms of which groups are scapegoated. Groups such as drug injectors, gay and bisexual men and sex traders are particularly likely to be targeted both by the scapegoaters and by HIV. Given this framework, how should public health professionals and activists engaged in HIV-related issues respond? Under what circumstances should we orient efforts upwards towards corporate, political or bureaucratic leaders? Under what circumstances, and how, should we orient towards popular forces? Relatedly, we need to consider an issue we often ignore: What do we have to offer potential allies? That is, in terms of their goals, philosophies and needs, why should they ally with us? PMID- 9743732 TI - Mental health problems in older adults with HIV referred to a psychological medicine unit. AB - Although HIV is still seen by many as a disease affecting younger adults, it is known that at least 11% of individuals with AIDS in Britain are over the age of 50. It is likely that this older age representation will continue and increase and whilst much is known about the psychological wellbeing of younger individuals with HIV, it is crucial that we consider the differing experiences and needs of older adults with the disease. Fifty-two adults with HIV over the age of 54 were referred to the Department of Psychological Medicine between June 1990 and December 1996. Data for these patients were compared with corresponding information for a random sample of younger patients with HIV. The older adults were found to differ significantly from the younger individuals on a variety of social, psychological and medical variables including social isolation, employment worries, sexuality, previous psychiatric history and stage of HIV at referral. It seems that current HIV services may actually alienate a significant proportion of potential users through not being sensitive to the needs and views of these older individuals. This must be addressed by policy makers and practitioners to ensure that psychiatric and psychological services become more acceptable and accessible to the older adult. PMID- 9743734 TI - Client preferences for HIV inpatient care delivery. AB - This study was concerned with preferences for inpatient models of care by the HIV/AIDS client group, in particular the difference between gay white men (European) and black heterosexuals of African/Caribbean origin. Satisfaction with the care currently provided was also an area of interest. Thirteen per cent (n = 79) of the were surveyed. Seventy per cent (n = 56) of the HIV/AIDS client group indicated a preference for a dedicated care model. Significant results were obtained demonstrating differences in the care model preferred by gay white men and black heterosexuals (p < 0.01). Gay white men were much more likely to state they would leave the trust to receive dedicated care (p < 0.01). Black heterosexuals were more likely to state that they would change treatment areas to avoid dedicated care (p < 0.01) Differences in concern about confidentiality were noted between the two groups. Confidentiality may be one of a number of factors influencing preference of care for African/Caribbeans and this needs to be studied further. The clients surveyed were not universally satisfied with the care they had been receiving. Following the results of the survey radical changes in the management of HIV inpatient care were made. PMID- 9743733 TI - HIV-associated dementia: clinical, epidemiological and resource utilization issues. AB - Among mental disorders associated with HIV infection, dementia is the one most likely to have a major impact on public health, both as a result of the high levels of individual disability, and the greater demand of health care resource utilization. Epidemiologic and economic impact of HIV-associated dementia needs to be estimated, in order to provide policy makers and health managers with the information required for decision making and resource allocation. An increase in HIV encephalopathy prevalence rates may be expected as a consequence of longer survival time in dementia patients and in patients with other AIDS defining disease (longer survival increases the risk of developing HIV encephalopathy). A resource utilization study shows that, in the chronic stage of the disease, in patient days per person-year are almost double in AIDS subjects with neurological complications as compared with those without neurological complications; no major difference appears when considering out-patients consultations and day-care treatments. In conclusion, a significant rise in resource utilization and in related costs may be anticipated as a consequence of the increasing prevalence of HIV encephalopathy. Further studies seem necessary to compare different approaches in the management of this debilitating disease, in view of a more rational utilization and allocation of resources. PMID- 9743735 TI - Breaking (through) the law--coming out of the silence: nursing, HIV/AIDS and euthanasia. AB - This paper provides a nursing perspective on ethical, legal, professional and practical issues faced by nurses working in HIV/AIDS care in relation to euthanasia/assisted suicide. Nurses who care for PLWHA (People Living with HIV/AIDS) have been conspicuously silent in the recent debates about euthanasia in Australia. Many factors prevent nurses from openly acknowledging their participation in assisted suicide/euthanasia and contributing to important debates about this topic. Their commitment to client confidentiality and the illegality of the practices are clearly significant factors which inhibit nurses from speaking freely. In addition, however, nurses' well-documented precarious legal position (Johnstone, 1994-alpha) and their subordinate status within the health care system make their public silence almost inevitable. Naming and challenging the factors which contribute to nurses silence, this paper draws on the experiences of nurses who have cared for PLWHA who have requested assistance in dying. It identifies practical, ethical and legal issues and dilemmas which can arise for nurses who may be involved in these practices, highlighting their special skills, relationships with clients, responsibilities and the complexity of their role; it also elucidates, however, the serious professional and personal risks nurses face given the legal and legislative status quo. This paper suggests that nurses may play a central, though covert, role in assisted suicide/euthanasia in HIV/AIDS care, rendering it imperative that their perspectives be included in the debates about the legalization of assisted suicide/euthanasia. Moreover, the paper identifies and challenges some severe impediments nurses must confront and address if they are to be able to contribute fully to this debate and to those which may arise in the future. PMID- 9743736 TI - Changes in patterns of risk. AB - Against the background of debate about the nature of risk in relation to HIV transmission, and resultant changes in risk discourse from risk group to risk behaviour to risk situation, declines in the velocity of HIV spread are being documented in countries with high levels of political commitment and multi sectoral approaches. Biological markers of sexual activity such as sexually transmitted disease incidence and HIV prevalence corroborate reports that young adults in Uganda and Thailand are increasingly adopting preventive behaviours. Risk perception, perceived social and community norms and self-efficacy influence behavioural change, but structural and contextual factors, including resource constraints affecting HIV prevention programmes and the treatment of sexually transmitted diseases, and social marginalization and stigmatization affecting access of vulnerable populations to the means of prevention play determinant roles. Both patterns of risk for HIV infection and our understanding of them are evolving. More research is required to gain knowledge and understanding of factors such as partner concurrency as well as cultural and socio-economic determinants and contextual underpinnings of sexual and drug injecting networks. This is an essential first step to mobilizing communities to take action at the individual, partnership and community levels to reduce risk. PMID- 9743737 TI - Perceptions of HIV/AIDS and caring for people with terminal AIDS in southern Thailand. AB - This study presents data collected from village-based ethnographic research conducted in southern Thailand in 1995-1996, and focuses on perceptions of HIV/AIDS infection, patients with AIDS and their provision of care. Individual interviews were conducted with 300 village women. These data were supplemented by data from 14 focus group discussions involving 100 participants, both men and women, randomly selected from six villages in Hatyai district, Songkla Province, Thailand. In addition, 23 people with HIV/AIDS and their caregivers participated in subsequent in-depth interviews. Participants generally obtained their information about HIV/AIDS from television and radio, and the information they obtained was generally negative. AIDS was perceived as a disease associated with dirt, danger and death, although it was also considered to be a disease of karma (rok khong khon mee kam) and a 'woman's disease' (rok phu ying) associated with prostitution. Few women perceived themselves to be at risk of infection because they 'trusted' their husbands to be faithful. There were some differences in attitudes towards caring for AIDS patients among people who lived in semi-urban and rural areas, and with areas which had not yet experienced AIDS among community members. Focus group discussions clarified issues related to the illness and patterns of caregiving among men and women. Areas of misperception and confusion were identified and will be used for interventions. PMID- 9743738 TI - Sexual practices among men attending an anonymous HIV testing site in Singapore. AB - This study was undertaken to provide an empirical basis for planning safer sex interventions in Singapore by examining the sexual practices of men attending an anonymous HIV testing site. All male clients attending the testing site from January, 1996 to January 1997 were asked to complete an anonymous questionnaire concerning oral and anal sex with men as well as vaginal, oral and anal sex with women within the past year. Questionnaires were obtained from 251 male clients, ranging in age from 17 to 70 years (mean = 30.3). Of these 50.6% reported sex with women only, 39% with men only, 7.2% with both men and women and 3.2% with no one. Condom use was moderately high for anal and vaginal sex but low for oral sex. A risk index based on the riskiness of specific activities showed that bisexuals were the most likely to engage in high risk behaviour whereas homosexuals were the least likely. One factor in the greater riskiness of bisexuals' behaviour appeared to be a higher frequency of unprotected sex with women. These findings provide a preliminary portrait of sexual risk-taking among men in Singapore and suggest the need for continued emphasis on consistent condom use for penetrative sex as well as appropriate precautions for oral sex. The results also suggest the need to develop targeted interventions for bisexuals, particularly with respect to unprotected sex with women. PMID- 9743739 TI - Body image and HIV: implications for support and care. AB - Very little formal research has looked at body image change over the course of HIV illness or assessed the implications of changes for support interactions. There are three main spheres of influence on body image: the physical, psychological and the social. HIV shares some of these aspects with other chronic or fatal illnesses, but has specific elements which are distinctive, such as particular physical manifestations and the negative impact of media, social representations and stigma resulting in a radically altered experience for an HIV positive body. This paper outlines preliminary findings using a body image measure designed specifically for use in HIV. The results suggest that people with HIV may experience significant feelings of contamination, brought about through internalization of stigma and representations, in addition to physical decline as illness progresses. PMID- 9743740 TI - Employment, accommodation, finances and combination therapy: the social consequences of living with HIV/AIDS in Australia. AB - The research reported here is of a study of the psychosocial impact of living with HIV/AIDS in Australia focusing on employment, accommodation and income in the environment of new treatments for HIV/AIDS. Many people experience profound changes to their lifestyle as a result of living with HIV/AIDS. In addition to detrimental changes in their health, many people experience major changes in their employment, accommodation, finances and relationships. The research highlights the significance of psychosocial factors along with changes in physical health in shaping PLWHAs (People Living with HIV/AIDS) changes in employment and accommodations. The new treatments now available for HIV/AIDS are further transforming people's attitudes, with many PLWHA considering returning to employment. PMID- 9743741 TI - The global epidemic. PMID- 9743742 TI - Life-threatening complications and irreversible damage following maxillofacial trauma. AB - Trauma remains one of the principal causes of mortality in the western world, especially among young adults. The most serious immediate life-threatening complication following maxillofacial trauma is airway obstruction. The onset can be sudden, as with foreign body aspiration, or following soft-tissue damage that can lead at a later stage to airway-compromising oedema. The medical literature regarding facial trauma appears to support the hypothesis that maxillofacial trauma alone is rarely life threatening, or will not lead to life-threatening conditions unless associated with airway compromise. There are some causes of life-threatening complications following trauma to the maxillofacial region such as massive bleeding or undiagnosed cervical spine injury. However, there are some situations that may cause irreversible damage unless immediate operation is undertaken. The almost complete lack of reports dealing with death or irreversible damage in trauma involving the maxillofacial region prompted us to review and analyse the importance of immediate intervention following trauma to the maxillofacial region, in order to treat life-threatening complications and prevent irreversible damage. PMID- 9743743 TI - The use of facsimiles in acute patient transfers: experience with brachial plexus injuries. AB - In order to optimize the transfer of patients with acute brachial plexus injuries a facsimile proforma was introduced. The transfer of 20 patients prior to its introduction was compared as a retrospective control group with a prospective analysis of 20 patients transferred after the introduction of the facsimiles. There were many fewer missed injuries, the patients were better investigated and it as easier to decide when to delay patient transfer. Subjectively there was a considerably easier passage of both medical and social information about the patients. The cost of the facsimile and its use is more than compensated by the savings in reduced impatient stay, the reduced need for urgent tests upon receipt of the patient, fewer phone calls and considerably reduced administrative workload. PMID- 9743744 TI - The Hyphecan cap: a biological fingertip dressing. AB - An occlusive biological dressing, the Hyphecan cap, was evaluated in the management of full-thickness pulp defects of fingers and thumb in 20 consecutive patients. The cap was applied directly to the tip of the injured finger and allowed to separate gradually over the course of several weeks as new skin regenerated. This approach proved simple and effective in achieving a good functional and cosmetic result within a relatively short time, and eliminated the pain and discomfort associated with conventional dressing changes. We recommend the use of this dressing material in the Accident and Emergency department for the out-patient management of de-epithelializing fingertip injuries in both children and adults. PMID- 9743745 TI - Subtalar instability following lateral ligament injuries of the ankle. AB - A stress radiograph of the subtalar joint was taken using Telos equipment, while the X-ray beam was directed onto the posterior subtalar joint at angles of 30 degrees latero-medially and 40 degrees caudocranially. From this radiograph the subtalar tilt angle was measured as an index of stability of the subtalar joint. Intra-and interobserver evaluations of measurement of this angle were performed on 20 unstable ankles. The errors at a 95 per cent confidence level were 1.9 degrees for intraobserver A, 1.4 degrees for intraobserver B and 2 degrees in interobservation. Stability of the subtalar joint was evaluated on 46 lateral ligament injuries of the ankle (23 acute injuries and 23 chronic injuries) and 80 normal ankles. The subtalar tilt angle was 9.7 degrees +/ 3.2 degrees in acute injuries 10.3 degrees #/ 2.9 degrees in chronic injuries, and 5/2 degrees +/ 2/6 degrees in normal ankles. There were significant differences between the acute or chronic injury and the normal ankles. These results suggested that stability of the subtalar joint was disturbed following acute and chronic lateral ligament injuries of the ankle. PMID- 9743746 TI - The treatment of selected fractures of the humeral shaft with the True-Flex nail. AB - The True-Flex nail was used in 23 selected non-pathological and eight pathological fractures/lesions of the humeral shaft. The overall fracture union rate was 69.5 per cent and the patients achieved a good range of movement of the shoulder and elbow. Nailing did not lead to union of established non-unions or fractures which were previously treated unsuccessfully by surgery despite bone grafting. All patients with pathological fractures/lesions regained good function of the arm and a good range of movement of the shoulder and the elbow. In one case the nail migrated proximally and impinged on the rotator cuff. This was revised. No other technical difficulties or complications were seen. The True Flex nail is useful in the treatment of difficult and relatively recent humeral shaft fractures. Established non-unions or cases where previous surgery has failed should be treated by alternative methods. PMID- 9743747 TI - Long-term physical, psychological and social consequences of a fracture of the ankle. AB - The long-term physical, psychological and social outcomes of 68 patients with an ankle fracture were investigated by using a postal questionnaire 6 years after injury. Patients were treated at a level I Trauma Centre between January 1989 and December 1989. Where applicable, the outcomes were compared with the outcomes of severely injured patients who were investigated previously. Physically, the patients were suffering from problems in the extremities and the spine. Surprisingly, half of them (52 per cent) had psychological complaints due to the initial injury. Eighty-nine per cent of the patients returned to work. This result seems to be only slightly better than the return to work in severely injured patients (74 per cent). Those with an ankle fracture needed less time to return to work (3 months versus 13 months in the severely injured patients). Further social changes (marital status, leisure activities) were mostly found in the severely injured patients. Patients with an ankle fracture as well as severely injured patients are affected by long-term consequences. The differences between the two groups are not as pronounced as is generally assumed. Probably, the consequences of lesser injuries are frequently disregarded, whereas severely injured patients are doing better than expected. PMID- 9743748 TI - Long-term physical, psychological and social consequences of severe injuries. AB - This 6 year follow-up study was designed to evaluate the long-term physical, psychological and social outcomes of severely injured patients (Injury Severity Score of greater than or equal to 16). Patients were treated at the University Hospital Groningen, the Netherlands, between January 1989 and December 1989. Outcomes were assessed using a postal questionnaire. After injury, the 55 respondents had predominantly complaints of the extremities, the spine and the head. Psychological complaints were present in 84 per cent of patients and mainly concerned fatigue, slowness and memory impairments. Despite these physical and psychological complaints, 74 per cent of patients were able to return to work and the memory succeeded in complying with job requirements. Injuries of the extremities and the spine were risk factors for failing to return to work. Social consequences were also reflected in broken marriages (6/22) and changes of leisure activities (45 per cent). On the basis of the impairments and disabilities revealed, we conclude that further improvement of the long-term outcomes of severely injured patients may be achieved by advancements in the treatment of injuries to the head, spine or extremities, comprehensive psychological support and vocational rehabilitation. PMID- 9743749 TI - Femoral fractures in the elderly treated with an unreamed titanium nail. AB - We present a review of 28 femoral fractures in 28 elderly patients treated with an unreamed titanium intramedullary nail (AIM femoral nail, ACE medical, Figure 1). The series included 11 male and 17 female patients with an average age of 74.5 yr (range 63.5-93 yr). One fracture was open and the rest, closed (six pathological). Average follow up was 6 months (range 4.5-21 months). Mean time to bony union was 19 weeks (+/- 3.5). The union rate was 91 per cent with two delayed unions; one was a subtrochanteric pathological fracture and the other a diaphyseal fracture. Shortening of 2 cm occurred in one patient and one had malrotation. There was no implant failure in our series either with the screws or the nails. We feel that titanium unreamed intramedullary femoral nailing is an effective way of treating subtrochanteric and shaft fractures of the femur in old and frail patients. PMID- 9743750 TI - The role of the anterior column of the acetabulum on pelvic stability: a biomechanical study. AB - A study on the effects of the anterior and the posterior column of the acetabulum on pelvic ring stability was carried out by applying a lateral compression force to specially prepared specimens. In group 1, the continuity of the right anterior column was disrupted by removing the acetabulopubic part of the anterior column; the posterior column was left intact in order to measure the strength of the posterior column. In group 2 the continuity of the right posterior column was disrupted by removing the ischioacetabular part of the posterior column leaving the anterior column intact in order to measure strength of the anterior column. The posterior column provided an average maximum strength of 759.43 +/- 229.51 N and the stiffness was 113.19 +/- 22.40 N/nm. The anterior column provided an average maximum strength of 2015.40 +/- 352.31 N and the stiffness was 301.57 +/- 98.67 N/mm. Thus the anterior column provides 2.75 times greater strength to the pelvic ring than the posterior column (P < 0.05). This finding may be important in open reduction and internal fixation of double column acetabular fractures. PMID- 9743751 TI - Safety standards for stab-resistant body armour: a computer tomographic assessment of organ to skin distances. AB - The protective properties of knife-resistant armour are quantified by the distance a test blade penetrates beyond a test sample into clay at a given energy. At present there are two proposed standards: penetration to 5 mm and penetration to 20 mm. Armour made to the higher standard specification (5mm) is necessarily heavier as it offers more protection. To determine the safety of these standards a retrospective review of 71 consecutive computerised tomographic (CT) scans was made. The minimum distance from the skin to the vital organs was measured. No organ would have been breached at 5 mm of knife penetration deep to body armour. 41% of pleurae, 61% of livers, 64% of femoral arteries, 25% of spleens and 6% of hearts would have been breached at a depth of 20 m of knife penetration. There was no significant difference in the minimum skin to organ distances between male and female subjects. The 20 mm standard does not offer adequate protection against knife attacks. PMID- 9743752 TI - Deaths and injuries due to unexploded ordnance (UXO) in northern Lao PDR (Laos). AB - Xieng Khousang province in Lao PDR (Laos) is one of the most heavily bombed areas on earth due to the secret bombing during the Vietnam war. This study presents the community-based cumulative incidence of injuries related to unexploded ordnance (UXO) after the war. The data were collected by one of the UK based non governmental organizations, Mines Advisory Group (MAG). Three districts in the province, reported to have the largest UXO load, were studied. Nearly half of the injuries and deaths involved children below age 15. The case-fatality rate was not different among children and adults. Males were significantly more likely to die of UXO injuries than females. We believe that UXO is an important public health problem in Laos for the following reasons: (1) 22 years after the end of the war, an average of one injury still occurs every other day (2) there is a high incidence among children below age 15 (3) the case-fatality rate is high (4) most injuries involve multiple fragments which usually require complex surgical and medical management skills. PMID- 9743753 TI - Pain clinic management of medico-legal litigants. AB - A preliminary study was undertaken to determine the relationship between a patient's compensation claim and their management in pain clinics. Fifty consecutive patients with compensation claims and undergoing treatment in local pain clinics were selected from the senior author's medico-legal practice. An initial postal survey of UK pain clinics was undertaken to establish the percentage of patients in pain clinics with musculoskeletal pain and also what proportion of clinics enquire as to the litigation status of their patients. Medical reports available from both plaintiff and defence were reviewed together with the hospital's and general practitioner's notes. Musculo-skeletal pain contributes on average 55 per cent of the pain clinic workload and the majority of pain clinics establish the litigation status of their patients. Of the patient series, the senior author assessed 98 per cent as having exaggerated symptoms in 44 percent and 20 percent were proven to be malingering via covert video evidence. The results suggest that many patients with compensation claims seek to cloak inappropriate symptomatology with verification and medical support in pain clinics. The wisdom and expense of treating such patients in pain clinics is questioned and the need for a nationwide study on this matter identified. PMID- 9743755 TI - Open reduction and internal fixation in 46 displaced intraarticular calcaneal fractures. AB - The subject of this report is the evaluation of 46 comminuted displaced intraarticular calcaneal fractures in 40 patients treated by open reduction and internal fixation. The results were validated by a score based on objective findings and a subjective gradation obtained by a visual analyzing score (VAS). The results were excellent or good in 30 treated fractures, while the results in 9 fractures were validated a satisfactory and in 7 as unsatisfactory. The results concerning disability and need for workman's compensation were promising in patients who had suffered fractures known to result in a high disability and compensation rate. PMID- 9743754 TI - Distal forearm fractures in children:the role of radiographs during follow up. AB - There is no consensus on the frequency of radiographic examination in the routine follow up of distal forearm fractures in children. This study was undertaken in an attempt to rationalize and optimize the use of ionizing radiation in these circumstances. The radiographs and clinical notes of 325 distal forearm fractures in children were retrospectively studied. Degrees of initial angulation were measured from all of the radiographs performed during follow up. Comparisons of outcome were made between the fractures with initial angulation under 10 degrees and over 10 degrees, types of fracture and the degree of reduction. Fractures with initial angulation of < 10 degrees had no clinically significant evidence of re-angulation and should be considered stable requiring only an initial diagnostic radiograph. Complete fractures, displaced fractures and fractures involving both the radius and ulna require more careful follow up. Residual angulation after manipulation under anaesthetic (MUA) of 5-10 degrees was not associated with an increased rate of re-angulation in this study. There is no apparent advantage in performing more than three radiographs in the majority of cases. The authors make recommendations concerning the optimal frequency of radiography in the follow up of forearm fractures in children. PMID- 9743756 TI - An intra-articular fracture of the scapulothoracic joint. PMID- 9743757 TI - Carotoid artery thrombosis following a penetrating oro-pharyngeal injury of unusual aetiology. PMID- 9743758 TI - Paget's disease and odontoid peg fracture: a case report. PMID- 9743759 TI - Flexibility training and flexible working in the European Union. PMID- 9743760 TI - Changing the attitudes of 'tomorrow's doctors' towards mental illness and psychiatry: a comparison of two teaching methods. AB - The General Medical Council's document 'Tomorrow's Doctors' (1993, GMC, London) recommended major changes in the undergraduate curricula of UK medical schools. In Nottingham, the fourth-year psychiatric attachment became shorter in duration, and interactive, problem-oriented, workshop-based learning replaced lectures. We compared the efficacy of this new teaching style in changing medical students' attitudes towards psychiatry and mental illness with that of old-style, didactic, lecture-based teaching. On the first and last days of their psychiatric attachment, 110 fourth-year-medical students (45 old curriculum; 65 new curriculum) completed two self-administered attitudinal measures: the Attitude to Psychiatry Questionnaire (ATP-30) and the Attitude to Mental Illness Questionnaire (AMI). We found that students had favorable attitudes towards psychiatry and mental illness before the attachment. These attitudes became more positive after the attachment in students from both curricula, with no significant difference between the groups and no gender difference. Students found patient contact rewarding, become more accepting of community care, and had greater appreciation of the therapeutic potential of psychiatric interventions. The interactive, student-centred, problem-oriented teaching of the shortened new curriculum appeared as effective in changing medical student' attitudes as a longer attachment with traditional teaching. PMID- 9743761 TI - Using attitude surveys in medical ethics research and teaching: the example of undergraduate willingness to treat HIV-infected patients. AB - As a break from the usual lecture or case discussion to teach medical ethics, this paper reports on the utilization of attitudinal surveys in the ethical areas where students face a choice of behaviours. The technique can stimulate students to better understand their own attitudes and hopefully motivate them to overcome resistance to more ethical but perhaps less desirable behaviours. However, the technique also serves as the basis for empirical ethical research. The first selection of the paper provides background on the conceptualization of attitudes, their measurement, and the statistical techniques for their analysis. In the second section, the special characteristics of medical students as a study population and ethical considerations in surveying them are discussed. Thirdly, the author uses his work in researching and in teaching about students' willingness to treat patients infected with HIV as an example of survey and analytical techniques. Finally, methods for using surveys in the classroom are presented. PMID- 9743762 TI - Changes in medical education: the beliefs of medical students. AB - This study aimed to explore medical students' beliefs about medical education in the light of recent General Medical Council (GMC) recommendations for change and to examine how these beliefs relate to the students' year of training. All undergraduate medical students at a London medical school were given a questionnaire concerning basic sociodemographic information and levels of agreement with a number of statements relating to medical education. The response rate was 75.4% (n = 383). Most students agreed with the majority of recommendations for change. However, two key recommendations, 'more community based teaching' and 'more optional courses', were supported by only 50.2 and 46.3% of respondents, respectively. Using factor analysis, students' responses were classified into five educational belief orientations relating to 'psychosocial' (e.g. communication skills), 'scientific' (e.g. new technologies), 'active' (e.g. optional courses), 'reform' (e.g. decreasing factual load) and 'group' (e.g. small-group teaching) educational belief orientations. The results showed variations in students' belief orientations across the 5 years of training. The results are discussed in terms of their implications for implementing the GMC recommendations and the impact of medical education on students' belief systems. PMID- 9743763 TI - Medical students' attitudes to the physician's oath. AB - Students at the Ben-Gurion University medical school take the physician's oath at the beginning of their studies. Student attitudes towards the content, timing and relevance of the oath were examined before the ceremony, 3 months later and in the fourth and sixth years. Eight-seven percent of the students were positive about taking the oath, most commonly because the oath represented being part of a medical team that is bound by behavioural norms. Forty-three percent supported giving the oath legal force. There was a progressive decline between the first and final years in positive attitudes towards the oath. The timing most favoured for the oath was the beginning of clinical studies. Three months after the oath ceremony only 18% of the students were able to cite three obligations from the oath. Three percent of the students felt that the oath would affect their behaviour. In students, eyes the oath seems to be an emotionally important ritual, whose value probably transcends its actual content. PMID- 9743764 TI - Developing a medical school alcohol policy. AB - The health and lifestyle problems of medical students and doctors give cause for concern on several fronts. We have developed a health policy for staff and students in Newcastle Medical School. This policy presents integrated recommendations relating to six key areas, i.e. alcohol, physical activity, sexual health, stress, occupational health and safety, and diet. The methods used to develop recommendations in relation to alcohol are described here. They were adopted by the working groups developing recommendations in the other areas. There were four key stages to policy development: establishing an information base; preparing a draft policy for consultation; consulting staff and students, and finalizing the policy. Consultation was a slow and challenging process but led to substantial revisions to the policy, enhancing its acceptability and likely success. The final policy includes a 3-year implementation plan setting out actions, resource implications and key players. Our policy, which has been adopted by the medical school and will soon be implemented, includes recommendations regarding changes to the school environment, training and education needs for staff and students, and access to services for those with alcohol related problems. Health policies should be developed in other medical schools and our approach offer a possible model. PMID- 9743765 TI - The views of senior students and young doctors of their training in a skills laboratory. AB - The study was carried out in the Faculty of Medicine and Health Sciences (FMHS), United Arab Emirates (UAE) University, UAE, where a skills laboratory was set up in 1988 to train medical students in clinical skills before they use such skills on patients. The students learn clinical skills using simulated patients, models and mannequins. The training starts in the first year of the 6-year undergraduate curriculum and continues until the end of the fourth year, after which students rotate through the clinical specialties. This study sought to identify: (1) the views of senior clerks (sixth year) and interns (first postgraduate year) regarding the clinical skills training in the skills laboratory (SKL) during the early years of the medical curriculum; (2) the differences in views between senior clerks and interns; (3) the differences in views between women and men students. Students' views about communication, interviewing, physical examination, therapeutic, diagnostic and laboratory skills were investigated. The results indicated that prior training in clinical skills was helpful to the senior clerks and the interns. Statistically significant differences in opinion were noted between the senior clerks and the interns, the interns being more positive about the usefulness of the SKL training compared with learning clinical skills directly on patients. There was no difference between men and women students except that women students were not comfortable with allowing their peers to examine any part of their bodies. The findings of the study have direct implications for the SKL programme. PMID- 9743766 TI - An investigation of the difficulty of computer-based case simulations. AB - This study investigated the characteristics of computer-based case simulations (CCS) that may be associated with case difficulty. Difficulty was defined as the average rating by physicians of examinee performance on a nine-point scale or the passing rate on the cases. Two data sets were used, one from an administration of 18 cases, the other from an administration of 22 cases with 13 cases used on both occasions. Stepwise regression procedures were used separately for case properties and for analytic scoring of key variables to identify the best sets of predictors of case difficulty. Because of the small number of cases, regression results were evaluated for consistency across both data and both difficulty measures. For key variables, the best set of predictors included the number of different serious errors of commission, risk items, and benefit items. In general, cases were more difficult for higher values of these variables. For case variables, the only consistent variable was the length of the paragraph that provided patient history, with longer paragraphs associated with more difficult cases. Other variables were less consistent, but were often related to the structure of the simulation or the severity of the patient condition. Although the findings for case variables were limited, the analyses were very helpful in illuminating the interconnections among the variables within cases. PMID- 9743767 TI - Impediments to bed-side teaching. AB - Bed-side teaching is the process of active learning in the presence of a patient. A cross-sectional study was conducted in a teaching hospital to obtain the opinions of clinical teachers about bed-side teaching including perceived hindrances to its implementation. Of 152 teachers, 78% responded to the questionnaire. Ninety-five per cent reported that bed-side teaching is an effective way to teach professional skills. Time constraints, noisy wards and patients not being available were reported as the most frequently experienced hindrances to bed-side teaching. The survey found strong support for bed-side teaching but a substantial number of barriers to its implementation. Further research is required to study methods that will improve bed-site teaching. PMID- 9743768 TI - Computer-based management of content in planning a problem-based medical curriculum. AB - The Faculty of Medicine at the University of Sydney has undertaken a major educational change from a traditional didactic 6-year, undergraduate entry programme to a 4-year problem-based programme to which only graduates are admitted. We have used two computer-based tools which proved invaluable in developing and managing the content of the new curriculum. The first, developed using a commercial database and made available on the Faculty's Intranet, provided a means for eliciting appropriate problems, organizing content fields and searching the information. The second, based on a spreadsheet, provided a means of displaying agreed content on implementation grids, both for self directed learning and conventional teaching sessions. Both provided ready access for scrutiny, interactions, review and planning by staff and they greatly enhanced the process of understanding the nature of the new curriculum, and thus in reassuring staff about the change. By merging the two tools, a definitive curriculum database is emerging. PMID- 9743769 TI - A randomized controlled study of portfolio learning in undergraduate cancer education. AB - Portfolio learning has not previously been reported for clinical undergraduate teaching. This open randomized study aimed to assess the effect of portfolio learning in the teaching of oncology to medical students. The project aimed to provide the student with a holistic understanding of the impact of the disease and its treatment on the patient and family, and the natural history of malignant disease, through long-term personal experience of a cancer patient. All undergraduate medical students entering Clinical Studies in October 1992 at the University of Wales College of Medicine were randomized to a study or control group. Both groups continued with the standard curriculum. Each study-group student followed a patient with cancer for 9 months, supported by bi-monthly small-group tutorials. Tutors were either general practitioners or hospital consultants, not necessarily oncologists; each was supplied with a tutor's resource pack of key oncology review papers. Students recorded triggers to learning and key items in a personal learning portfolio. Students' performances in clinical examinations and the contents of their portfolio was assessed. Final assessment was by hidden questions in the objective structured clinical examination (OSCE) in the final degree examination, when students in the study group showed higher marks in factual knowledge of oncology, particularly amongst the weaker students (P = 0.01). Those submitting portfolios for formative assessment had higher overall marks than those in the study group who did not (P = 0.04), representing the more motivated students. The whole study group showed a beneficial trend in their knowledge of oncology. PMID- 9743770 TI - Supporting Pre-registration House Officers: the needs of educational supervisors of the first phase of postgraduate medical education. AB - The GMC's publications The New Doctor (GMC 1997) outlines an approach for the education of Pre-registration House Officers using designated educational supervisors. In order to establish appropriate means of training consultants to undertake this role, the Centre for Postgraduate and Continuing Medical Education at the Queens Medical Centre, Nottingham undertook a needs analysis. This consisted of a series of individual interviews with directors of postgraduate education in the mid-Trent region, four focus groups with consultants, and two dissemination workshops. The findings from the research have led to the development of a modular educational programme, based on the perceived needs and optimum structure indicated by the participants in the research process. PMID- 9743771 TI - Evaluation of a training package in the assessment and management of depression in primary care. AB - This study aimed to evaluate the impact on the behaviour and attitudes of experienced general practitioners of a 10-hour training package in the assessment and management of depression. Twenty general practitioners participated. Both subjective and objective assessments were carried out which suggested significant improvements in both assessment and management skills. However, subjectively reported changes were not always supported by the objective data obtained from rating role-played interviews. The role-played patients rated the doctors as better communicators after training. All participants felt attending the course was beneficial. They all felt more confident in their abilities to deal with depression and said the skills they had learnt on the course would be useful to them in their future work. An outcome study is now underway in order to assess whether the training package, which has been demonstrated to have an impact on the behaviour, skills and attitudes of doctors, has an impact on the health of patients. PMID- 9743772 TI - The development of general practice standards in Australia. Royal Australian College of General Practitioners. AB - The Royal Australian College of General Practitioners has spent 4 years developing a set of entry standards which define the minimum features of general practices expected for the mid-1990s. The project design followed a slow, iterative process, with several opportunities for wide consultation with professional, consumer and Government groups. The draft standards were piloted in 25 volunteer practices, modified and then field-tested in 200 randomly selected practices representing urban and rural practices. Results of this field testing showed that the standards had content validity and that reliable measures were possible using triangulation from several data sources. The current version of the standards has been distributed widely for voluntary application in Australian general practices from early 1997. PMID- 9743773 TI - World Federation for Medical Education (WFME) Guidelines for Using Computers in Medical Education. PMID- 9743774 TI - Re: Dokuz Eylul joins the family: active medical education. PMID- 9743775 TI - An international database on medical education: the European Medical School Information System (EMSIS). AB - An electronic database containing the curricula and other information of more than 200 medical schools from 40 different countries has been compiled and made accessible via the World Wide Web (http:@yi.com/emsis), with the aims of providing an overview and to enable a comparison of the curricula of Medical Schools in Europe, of facilitating the cooperation between Medical Schools in Europe and allowing medical students to plan their studying period abroad. The database contains the addresses of medical schools, general information about each medical school (number of students, hospital beds available for teaching, etc.), exchange information (exchange networks the medical school is participating in) and extensive curriculum information, consisting of more than 10,000 course descriptions. All information has largely been entered by the medical schools themselves. Medical schools may update the database directly via the Internet at any time. Users may perform simple queries, for example by country or city, or may perform more complex searches such as 'show all medical schools teaching gynaecology in the fourth year'. PMID- 9743776 TI - Structuring ward rounds for learning: can opportunities be created? AB - This paper explores the ways in which ward rounds can be conducted to maximize educational opportunities, as part of a project to improve the effectiveness of on-the-job training (OJT) for hospital doctors. Ninety ward rounds taken by 24 trainers in the Anglia region were observed. Each observation produced a note of the ward round's structure and routines and of the contributions made to it by trainers and trainees. Teaching was a feature of all ward rounds and different types of rounds were valued for different reasons. A range of ward round structures was observed and, within each, a range of routines for conducting the round. Ward round structures fell into four categories, with almost three quarters of trainers making no use of either pre- or post-round meetings. Where such meetings took place, however, opportunities for OJT were created and, in some cases, optimized through routines to encourage trainee contributions. Discussion time away from patients structured into ward rounds enabled trainers and trainees to take advantage of many opportunities to learn from service. Although unplanned and unsystematic opportunities for OJT do arise, far more reliable are those created through systematic planning and preparation. Trainers have choices to make about how they conduct ward rounds and by choosing to make use of pre- and/or post-ward round sessions, valuable opportunities for OJT can be created. PMID- 9743777 TI - Are vocationally trained general practitioners better GPs? A review of research designs and outcomes. AB - Implicit and explicit in reviews of and changes to vocational education for general practitioners in the 1990s is the challenge to defend the assumption that vocationally trained GPs are better GPs. This paper provides a review of the international literature which has reported on outcomes of general practice vocational training programmes. Through the review we identify both the types of research methodologies used (including a brief discussion of their strengths and limitations) and the outcomes reported of vocational training. Twenty-five studies on the outcomes of vocational training are reviewed. These studies used multiple data sources and one of four methodologies: pre- and post-training comparisons, analysis of learners' or teachers' accounts, audits of general practice or analysis of examination pass rates. When collated, the following range of outcomes from vocational training were identified: improved quality of patient care, increased knowledge, improved general practice skills, increased confidence and desirable GP attitudes and personality traits, increased adherence to practice guidelines and higher examination pass rates. The paper concludes with a summary of research and education issues which arise when we examine the question posed at the outset: are trained GPs better GPs? PMID- 9743778 TI - The relationship of tutors' content expertise to interventions and perceptions in a PBL medical curriculum. AB - This study addressed three questions: (1) Do content-expert tutors differ from non-expert tutors in the extent to which they present/explain case content? (2) Do tutors who present/explain case content differ from those who almost never do in their ratings of various outcomes of a PBL curriculum? (3) Are tutors who present/explain case content rated differently by students from tutors who almost never do? Data were gathered from 88 tutors and 168 students in the first 2 years of a PBL medical curriculum. Students assessed their tutor after each unit, and tutors completed a questionnaire near the end of the academic year. In this study, 'content expertise' was defined operationally as tutors' self-ratings on the question 'To what extent could you teach (at the Med I level) the material covered in the cases?' Less than half of the tutors reported that they almost never presented/explained case content. As tutors' content expertise increased, they tended to present/explain case content more frequently. Tutors who almost never presented/explained case content rated PBL more highly than traditional methods. No differences were found in student ratings of tutors who almost never presented/explained case content, compared to tutors who did. The results suggest that tutors who are content experts find it difficult to maintain the 'facilitator' role, but that those who maintain this role are more satisfied with PBL. It appears that other tutor behaviours may have a greater influence on students' ratings of their tutors. PMID- 9743779 TI - Tutor intervention profile: reliability and validity. AB - An instrument has been developed to provide tutors with feedback about their performance in problem-based tutorial groups. This questionnaire consists of 33 statements reflecting the behaviour of the tutor in the tutorial group when students analyse the case and generate learning issues and the behaviour of the tutor when students report back to the tutorial group about their self-study. The statements are grouped on theoretical dimensions found in the literature about co operative learning and problem-based learning. This study focuses on the validity and the reliability of this instrument. Confirmatory factor analysis showed that a four-factor model showed a weak fit, whereas each separate factor fitted the data reasonably well. The four factors are: (1) elaboration; (2) directing the learning process; (3) integration of knowledge and (4) stimulating interaction and individual accountability. These factors were strongly correlated with the overall judgement of the tutor at the end of the unit, so the concurrent validity of this instrument was high. Generalizability studies indicated that the rating scales provide reliable information with a sample size of student responses falling within the range of real tutorial group sizes. The results of this study show that the instrument for providing tutors with feedback is valid and reliable. This questionnaire can be used in staff development activities and also provides needs assessment for faculty development. PMID- 9743780 TI - The impact of an individual tutorial session on MEDLINE use among obstetrics and gynaecology residents in an academic training programme: a randomized trial. AB - Over the past decade, on-line databases have become increasingly popular among health care professionals. As a group, these 'end-users' report utilizing databases to keep abreast of medical progress, to conduct research and to address specific patient care issues. Throughout the literature, medical professionals ('content experts') have proved to be less effective searchers than librarians ('search experts'). The potential implications of this discrepancy are worrysome. For any given clinical scenario, for example, published reports may reach contradictory conclusions. A poorly skilled searcher may not retrieve enough articles to appreciate this fact. Optimizing searching skills is therefore a worthwhile goal. As a first step, many medical schools introduce students to on line databases, most notably MEDLINE. Residency is an ideal time to continue this training. A recognized obstacle to provide residents with formal MEDLINE instruction is time constraint. We therefore conducted this study to ascertain the impact an individual 1-hour tutorial session would have on MEDLINE utilization among obstetrics and gynecology residents training at an academic medical centre. Outcome measures included MEDLINE search frequency, duration, recall, precision and searcher satisfaction. Search recall measures the searcher's ability to retrieve articles deemed relevant to the question at hand. Search precision gauges the searchers' ability to eliminate irrelevant articles. Although the sessions were well received, we were unable to demonstrate an improvement in the outcome measures analysed. Further research is therefore indicated so that cost-effective educational strategies can be recommended for wide-scale use. PMID- 9743781 TI - Latin American biomedical publications: the case of Colombia in Medline. AB - Latin America generates a low proportion of the references quoted in Medline, the most popular health-related literature search database in the world. This paper explores references from and about Colombia in Medline during the period 1987 1996. Topics addressed, patterns of authorship and research locations are established. The number of Latin American journals indexed in Medline has been progressively reduced during this 10-year period, with Colombian journals completely excluded since 1991. During this 10-year period, the total output of Colombian research institutions in foreign journals consisted of 531 articles, 41% (219) of which come from the four leading universities. These figures are substantially lower than those from other countries of the region such as Venezuela or Chile. Despite some governmental efforts, Colombia continues to have a low scientific output and has yet to attract the interest of foreign researchers. Alternatives for development of Latin American research and publications are offered. PMID- 9743783 TI - The rise and fall of students' skill in obtaining a medical history. AB - A gradual shift towards a more humanistic conception of medicine has occurred in recent years. Along with this shift have come attempts by medical educators to include interviewing and communication skills as part of the medical curriculum. The current study evaluates the effectiveness of a clinical medicine curriculum which emphasizes interviewing skills. Between 1992 and 1994 and 292 graduates of the University of Connecticut School of Medicine participated in five clinical skills teaching and assessment programmes during the four years of medical school. During each of these five programmes, the students' interviewing skills were rated using the Arizona Clinical Interview Rating Scale (ACIR). The raters were standardized patients with whom they had just completed a medical encounter. Results show that students' development of skills differed, with closure items showing the greatest increase and social history items showing the greatest decline, with an overall initial increase and then a decline in interviewing skills over the four years. Explanations for these findings include the de emphasis of communication skills during the clinical years and the culture of medicine to which students are exposed during these years. PMID- 9743782 TI - The use of multidisciplinary consensus groups in the planning phase of an integrated problem-based curriculum. AB - In response to the General Medical Council's initiative to reform UK medical undergraduate education only a minority of medical schools have developed entirely novel curricula. Although the experiences gained by these schools in curriculum design and planning have not been recorded in the literature they are likely to be of interest to other medical schools still contemplating course revision. The medical school at the University of Liverpool recently launched an integrated problem-based course differing radically from its predecessor. The General Medical Council considered integration of contributing disciplines one of the most important aims of reform, yet courses that integrate independent component disciplines may be perceived by staff as threatening due to the loss of structure and disciplinary autonomy. Strategies for early course development must take account of these concerns as well as dealing with the identification of course content. A multidisciplinary consensus group process, designed to combat some of these problems, was employed to identify the learning objectives and core content for the new course. The purpose of this paper is to describe, first, the processes employed to identify the core palliative care component for a new PBL curriculum and secondly, how these objectives were integrated horizontally and vertically throughout all course elements. PMID- 9743784 TI - A prospective randomized trial of an urban general practice attachment for medical students. AB - A randomized controlled trial was undertaken to evaluate the effectiveness of a new model for providing urban general practice attachments for final-year medical students at the Flinders University of South Australia. All the student groups in that year were randomized prospectively to either the standard student attachment, as run by the university, or to an attachment organized by a project team from a local network of general practitioners. Students in the intervention group had their personal learning goals assessed and matched with their general practice preceptors, and the students were set a task that developed their contact with other health resources in the community. Results from an evaluation questionnaire completed by the students at the end of their terms showed that the students in the intervention group rated their general practice preceptors more highly, had more contact with allied health and community organizations, felt that they had met their own learning goals to a greater extent, and enjoyed their term more. Student examination results showed that the students in the intervention group did not perform as well in one of the four areas of their end of term examination as did the students in the standard attachment. The additional cost of providing the intervention was estimated to be A$340 per student. We conclude that long-term decisions about adopting this new model of organizing general practice attachments on a wider scale will need to balance the apparent benefits against the increased resources required. PMID- 9743785 TI - Grade predictions for school-leaving examinations: do they predict anything? AB - The attributes of 721 medical students admitted to the United Medical and Dental Schools (UMDS) of Guy's and St. Thomas' Hospitals, London, UK between the years 1991 and 1994 were examined to determine the relationship between A-level grade predictions (as completed on each student's UCAS form by their school/college at the time of their application to the school) and subsequent assessments of their academic ability and performance. Predicted A-level grades were found to be significantly, if weakly, correlated with the rating of academic ability made at interview by the UMDS interviewing panel. They were not however, related to the grades obtained by students in the A-level examination itself. Further, while success at pre-clinical examinations was predicted by obtained A-level grades, it showed no relationship with the predicted grades. In contrast, the interview based rating of the applicant's academic potential was significantly predictive of subsequent A level and, to a lesser extent, pre-clinical examination performance. It is concluded that predicted A-level grades may not offer a valid method of assessing the academic potential of applicants for medical school. PMID- 9743786 TI - Undergraduate teaching in a day surgery unit: a 2-year evaluation. AB - Medical schools are having to adapt their teaching in response to the reduction in inpatient availability and the increase in outpatient and community care. A surgical course for first clinical-year undergraduate medical students was established in the day surgery unit at King's College School of Medicine and Dentistry in 1995. It was considered desirable because of the shift to day-case surgery, and proved feasible to run (Seabrook et al. 1997a). The course is taken by one-third of the annual intake of 120 students. A formal evaluation was undertaken comprising comparison of student performance in end-of-year clinical and multiple choice examinations and of acceptability to students, teachers and managers. The results showed no significant differences in performance between students who had taken the day-surgery course and those on other surgical attachments. Students' satisfaction with the course was significantly higher on nine individual criteria and lower on four criteria. The teaching was positively viewed by teachers and managers, despite having costs as well as benefits. We conclude that day-surgery centres can be used successfully to teach medical undergraduates. Experience in both inpatient and day-case environments should benefit students by reflecting more closely the reality of current surgical care. PMID- 9743787 TI - Shared goals, shared learning: evaluation of a multiprofessional course for undergraduate students. AB - Twenty-eight undergraduate degree students from seven health care professions attended a two-day pilot course. Using small multiprofessional groups, final-year students from occupational therapy, orthoptics, therapy radiography, nursing, physiotherapy, medicine and dentistry explored professional roles and clinical problem-solving using a theme-based approach. A balance of didactic and interactive small-group learning enabled them to identify issues surrounding multiprofessional teamworking and collaboration in the National Health Service. Evaluation results showed that the course increased knowledge and understanding of other health care professions, developed more positive attitudes and demonstrated the importance of multiprofessional teamwork and communication. Participating students believed that both early and regular opportunities for shared learning should be essential aspects of undergraduate courses. PMID- 9743788 TI - Measuring the hospital experiences of junior doctors. AB - The development of an appraisal questionnaire which measures junior doctors' opinions about their hospital experiences is described. The first section of the questionnaire consists of seven reliable subscales which measure opinions about teaching and learning, registrar teaching, consultant teaching, staff support, workload, administration and overall experiences during a period of attachment or term. The second part of the survey contains 11 reliable questions about the hours spent on service and education during the term. The responses to this measure of 257 randomly selected Australian junior doctors are described. The questionnaire may be used to contrast the experiences of junior doctors in different types of terms, different hospitals or varying levels of training. The data generated from the instrument can provide useful information about hospitals, such as the work practices of junior staff and the effectiveness of educational programs. PMID- 9743789 TI - Expectations and opinions of pregnant women about medical students being involved in care at the time of delivery. AB - Information was collected from pregnant women about their knowledge of and previous experience with medical students; their opinions towards medical students being involved at the time of delivery and the socioeconomic and religious influences on these. The format of the study was a self-administered questionnaire survey. The setting was a teaching hospital in the UK. In total, 118 pregnant women aged between 15 and 46 years, with a gestational age of 18-42 weeks were surveyed. Factors which significantly influenced acceptance of medical students were found to be previous number of children (P = < 0.001) and religious beliefs of the pregnant women (P = 0.002). Only 51.4% of antenatal women knew that a "medical student" is a doctor in training and most assumed that the role of the student at the time of delivery required very few clinical skills. Only 13.6% knew that medical students could deliver a baby under supervision. Of the 118 subjects, 95.4% thought that student participation at the time of delivery was a worthwhile learning experience; however, only 74.6% were actually willing for a student to be involved. In conclusion, pregnant women appear to have made their decisions about medical student participation by balancing personal needs with a sense of responsibility to help in the education of others. The results suggest the patients need more information about medical students, including an explanation of the term 'medical student' and an outline of the role they play during the intrapartum period. PMID- 9743790 TI - An update on British medical students' lifestyles. AB - Information about medical students' lifestyles was obtained from 785 second-year students from seven medical schools in Great Britain by a personally administered questionnaire. Fifteen per cent of the students were non-drinkers. Among those who drank, 48% of the men and 38% of the women exceeded sensible weekly limits of alcohol consumption, and high-risk levels of consumption were reported by 12% of men and 7% of women. Cannabis had been used at least once or twice by more than half the men and 40% of the women, and 10% reported regular use (weekly or more often). Experience with other illicit drugs was also reported: amphetamines (8% of students), LSD (7%), ecstasy (4%), amyl/butyl nitrate (10%) and magic mushrooms (7%). Nineteen per cent of the students had used two or more different drugs. Experience with illicit drugs started before entering university in more than a third of those who used them. Comparison of the results with other student surveys suggests that the lifestyles of medical students differ little from those of other student groups, but that alcohol and illicit drug consumption is increasing in university students generally. Prospective studies are under way to establish whether medical students change their lifestyles at later stages of their course and after qualification. PMID- 9743791 TI - Video recording feedback: a feasible and effective approach to teaching history taking and physical examination skills in undergraduate paediatric medicine. AB - Medical educators have always recognized the need to teach and train medical graduates and undergraduates the skills of conducting a consultation. Several authors have established the efficacy of using constructive feedback on videotape of each student's interaction with a patient to teach and enhance such skills. This study reports 'students' perceptions' of the feedback process used in the Junior Paediatric Clerkship at the Faculty of Medicine of the United Arab Emirates University. An unexpected 73% of the respondents believed that self observation influenced development of their clinical skills. More than 80% said that the feedback from instructors and peers helped them to improve their clinical skills, but they would have liked to have more than one of their consultations recorded and reviewed. It was found that 75% of the students felt that self-critique of their performance made them aware of their strengths and weaknesses and their skills in analysing and evaluating consultations had been enhanced. It was found from Kruskal Wallis one-way ANOVA that the students' professional attitude, empathy, and warmth towards the patients differed highly significantly (P = 0.0062, 0.0089, 0.0007, respectively) from self-assurance, self-confidence and competence. They were also deficient in certain areas of history-taking, interviewing skills, and physical examination techniques and perceived they needed more training in order to be proficient. PMID- 9743792 TI - Assuring specialist practice. PMID- 9743793 TI - Imprecise terms in UK medical multiple-choice questions: what examiners think they mean. AB - Many multiple-choice questions (MCQs) used in medical education in the UK contain undefined, imprecise terms. They are particularly common in true/false items and can be found in classroom tests, published examples of MCQs and, more importantly, in high-stakes examinations which determine a candidate's graduation or membership of a professional body. This study investigated imprecise terms used in some MB BS final examinations and the Part 1 Membership Examination of the Royal College of Physicians. It revealed that imprecise terms occur commonly, yet there is a wide range of opinion among the examiners themselves about their meanings. The numbers and variety of imprecise terms which were found in high stakes MCQ examinations are described in this paper and details are given concerning the lack of consensus about their meanings as reported by the responsible examiners. A second type of construction error--disproportionately large numbers of 'true' branches--was also recorded. Exemplary practices do exist in MCQ quality assurance, but in the UK they are very much the exception rather than the rule. The findings of this investigation strongly indicate a need for change. PMID- 9743794 TI - Clinical teachers' perceptions of medical students' English language proficiency. AB - Medical educators from the Faculty of Medicine at the University of Adelaide, South Australia, have expressed reservations about the adequacy of some undergraduate medical students' English language proficiency for satisfactory academic and clinical performance. This study explores the occurrence and nature of the comments made in writing by clinical teachers about the English language proficiency of 568 students over a period of 4 years. The frequency and nature of the comments made by clinicians have important implications for the planning and implementation of pedagogical strategies to support non-English-speaking background medical students experiencing difficulties with their course due to language. Although the University of Adelaide has introduced initiatives in response to some of the problems that have been identified, it is recommended that any teaching interventions require careful evaluation through a longitudinal research design to ensure that their aims are being achieved. PMID- 9743795 TI - Auscultation of the heart: a trial of classroom teaching versus computer-based independent learning. AB - Declining skills in auscultation of the heart prompted an evaluation of teaching methods for medical students. A comparison of classroom teaching and computer aided independent learning of auscultation was carried out with two groups of approximately 20 second-year medical students. Both groups used approximately 20 recorded normal and abnormal heart sounds and murmurs, chosen to illustrate learning issues. For the classroom group a cardiologist presented each case through multiple stethophones and led the discussion. The individual study group used a new CD-ROM collection of cases and recordings in quiz format, with a hypertext link to a comprehensive text on auscultation and additional recordings. Students were tested with 16 multiple choice and 5 open questions on eight selected recordings, and evaluated the teaching by questionnaire. The classroom taught students scored higher on open questions than the CD-ROM-taught group, but in general performance by both groups was satisfactory and equivalent. Students of both groups repeatedly had difficulty classifying regurgitant and ejection murmurs and identifying characteristics of the second heart sound. Both CD-ROM and classroom teaching methods were highly rated by students but most students preferred a combination. PMID- 9743796 TI - Measurement of how well a paediatric training programme prepares graduates for their chosen career paths. AB - To provide a sound basis for modification of our paediatric residency education programme, we surveyed graduates from the past 16 years. The questionnaire was designed to determine the adequacy of training rotations in preparing graduates for their career paths. Questionnaires were mailed to 81 graduates; 73 (90%) replied. A modified version was completed by 27 of 29 current residents (93%). For most rotations, responses were normally distributed. However, 10 or more respondents identified exposure in one area as 'excessive' and in 6 as 'inadequate'. Current residents scored many rotations as 'inadequate', likely indicative of their limited exposure to actual practice. Recommendations were consistent for subjects needing more instruction. All major issues raised by graduates had been identified by faculty, but the substantiation enabled changes to be made with widespread support. We recommend periodic survey of graduates to evaluate how well education is preparing residents for their ultimate career paths. PMID- 9743797 TI - An evaluation of the 'short form' course experience questionnaire with medical students. AB - In this study we have validated the Course Evaluation Questionnaire (CEQ) as an instrument for use with undergraduate medical students. We have demonstrated satisfactory construct validity and reliability for the inventory. The CEQ is an appropriate instrument for course evaluation in medical education. PMID- 9743799 TI - Hospital doctors, general practitioners and dentists learning together. AB - The design, implementation and evaluation of an innovative course on education for a group of hospital doctors, general practitioners and dentists is described. It took place over five 2-day modules spread over 21 months. There were 33 participants and several tutors and external resources. The course was designed around group work and used a learner-centred agenda. Evaluation was by use of the nominal group technique during the course and by open questionnaires after 18 months. The course was successful in stimulating a number of activities and in developing the skills and confidence of the participants. Problem areas centred around the tension between the learner-centred agenda and the participants' need for structure. Courses such as this require good administration, as cohesive and supportive tutors' group, a clear statement of aims, even within a learner centred framework, and the establishment of clear ground rules to allow the participants to feel safe. Without safety it is arguable that less challenging learning will occur. PMID- 9743798 TI - Verifying the curriculum of a family medicine clerkship. AB - The learning experience during a medical school clinical rotation is largely shaped by students' patient encounters. This paper reports on how a log system for recording these encounters can be used for course planning and evaluation. Over the past 5 years, 960 third-year students completed log forms based on their clinical encounters during a required 4-week family medicine clerkship at UT Southwestern. These forms were then optically scanned and the information entered into a computerized database. Log form data revealed that the most common medical problems encountered by students in their ambulatory settings were similar to those reported in the general family practice literature. There was a great deal of consistency in the types of encounters from year to year. The data also showed some differences among clerkship sites in terms of patient demographics and the most frequently reported diagnoses. Information generated from student log forms has been used by the clerkship faculty to determine required readings, prioritize didactic topics and other teaching, adjust curriculum content, prepare support materials and develop examinations. Given the utility of the information obtained and the ease of use of optical mark encounter sheets, we recommend this system for other clerkships. PMID- 9743800 TI - Teaching problem-solving and clinical reasoning: 20 years experience with video supported small-group learning. AB - In the context of a curriculum reform the Faculty of Medicine of the University of Leuven, Belgium, introduced a new teaching project: video-supported small group learning on problem-solving and clinical reasoning. The aim of this study is to reflect 20 years experience. The video-supported sessions for sixth-year medical students during their practical year in peripheral hospitals were constructed in four stages. The first stage is the video presentation of a case with history-taking and physical examination. The student and three tutors of internal medicine make notes and can ask further questions and perform additional physical examination acts after the video presentation. The coordinator of the course, who knows the patient, then simulates the patient and the doctor to answer the questions. The second stage consists of making up individually a synoptic problem list, integrating history and physical examination; a differential diagnosis list with the most likely diagnosis fitting the problem list; and a list with investigations to be asked for confirming the diagnosis. The third stage consists of three small student groups discussing the three lists requested in stage 2. Each small group of students, passively assisted by a tutor, has to come to the consensus lists. The fourth stage is the confrontation of the consensus lists of the three groups with the aim of coming to an overall agreement. At this stage tutors are more actively involved in the discussion. Several learning processes are involved in this way of teaching. During the first stage the students learn the traditional teaching 'see one, do one and teach one', a demonstration of a full history and physical examination. By asking for additional information they learn by a critical attitude and by developing a strategy of fact-finding. During the second and third stages, by making their lists and during the consensus processes, they learn the significance of individual findings, problem-framing and the synthesis of history and physical examination data in medical concepts. The third and fourth parts of the sessions bring up the process of clinical reasoning, formulation of a working hypothesis, the discussion of the pathophysiology of findings, clustering of problems and epidemiological considerations as incidence and prevalence. Finally, the exercise to select diagnostic tests gives the students the possibility of appreciating the value of sensitivity/specificity and risks, benefits and costs of diagnostic procedures. These video-supported clinical problem-solving and reasoning sessions were positively appraised by students, teachers and medical faculty over the years. Over 20 years, more than 90 cases have been recorded on video, with a widespread variation in diagnoses and clinical presentations. Small-group teaching with the aid of a video case, as described in this paper, can promote enjoyable learning for students and teachers. PMID- 9743801 TI - Day surgery: teaching the next generation. AB - The advent of day surgery presents new opportunities and challenges for medical education. The opportunity to see patients pre-operatively and follow them through surgery to discharge on the same day is unique to day surgery. However, with rare exceptions, the development of educational programmes in ambulatory surgical settings is still largely at a rudimentary level. An undergraduate pilot programme was conducted at the University of Adelaide to explore the practicalities, acceptance and educational value of a day surgery programme for final-year medical students. The programme had three components: day surgery patient follow through, practical procedure tutorials and problem-based learning tutorials. It incorporated assessment of practical skills and theoretical knowledge with the use of log books and clinical and practical simulations as important elements in the assessment process. The pilot programme was accepted by all stakeholders and students' perceived significant gains in knowledge and skills. This programme may provide a teaching model that could be adapted for use in other medical schools. PMID- 9743802 TI - Planning and producing an educational videotape on the cervical screening programme in Wales. AB - Planning and producing an instructional videotape for health care workers require attention to educational principles but also depend on project management skills. The paper describes an approach to the planning and production of a programme on the cervical screening service in Wales that used focus groups to obtain information about needs assessment and developmental testing. Learning objectives need to be married with content accuracy and policy consensus, often difficult to achieve for health care areas such as cervical screening. For these reasons, effective and close liaison is required between sponsor, content experts, scriptwriter, producer and director. PMID- 9743803 TI - Portfolio learning in general practice vocational training--does it work? AB - Reflective learning has been widely addressed as an important learning mechanism in the educational literature. The creation of portfolios has been seen as a mechanism to promote this, though there has been little exploration of the place of a portfolio in general practice training. This study examined the introduction of a model of a portfolio learning strategy into one training region. The model had been developed by previous pilot work. The study explored the model in terms of its usefulness in general practice training and its relationship to reflective learning. An educational facilitator was used to support this introduction. Workshops and written material were developed to disseminate and refine ideas generated in the pilot study. This was followed by visits to trainer/general practice registrar (GPR) pairs over a 2-year period. These visits included semistructured interviews, which were tape-recorded and analysed using qualitative methods. Additional written resources, video and audio material as well as new workshops were designed with the researched participants in order to promote the development of the concepts of portfolio learning. Sixty interviews were carried out over a 2-year period with 44 pairs of trainers and GPRs. This included a total of 27 trainers and 44 registrars. Eighteen pairs were interviewed twice. Two focus groups were used at the end of the project. Portfolios have a place to play in general practice vocational training. They act as a bridge between hospital and general practice. They can be used to develop a learner-centred curriculum, explore difficult emotive concerns and facilitate feedback. They do not suit all learning styles. Their use is determined by a cost benefit analysis described in this study. PMID- 9743804 TI - Students' perceptions of problem-based learning at the B.P. Koirala Institute of Health Sciences, Nepal. PMID- 9743805 TI - Self and tutor evaluations in problem-based learning tutorials: is there a relationship? AB - It is now recognized that acquiring specific skills in self-evaluation helps students in a more general appraisal of their overall performance. Self- and peer evaluation skills are essential prerequisites for the success of every doctor in maintaining professional competence. In the Faculty of Medicine and Health Sciences (FMHS) of the United Arab Emirates, problem-based learning (PBL) is instituted in the first year. Self-evaluation by students and tutor rating of students' performances are an integral part of the PBL tutorials. This has provided the opportunity to conduct a systematic study of the role of self evaluation by students as distinct from the tutor evaluation of students in the PBL tutorials for five themes. The study sample included all preparatory year (first-year) students who joined the FMHS in 1994 and 1995. A total number of 64 students participated, of whom 17 (26%) were male and 47 (74%) female. Mean self evaluation scores were high throughout the module and did not follow any trend from theme one to theme five. While self and tutor scores were similar, male student self-evaluation scores were higher than for female students on overall scores. The sharing of assessment reports between students and tutors has been perceived to be a useful tool for the students' development of the skills of analysis, differentiation and critical appraisal. PMID- 9743806 TI - Who should assess medical students' communication skills: their academic teachers or their patients? AB - The objective of this study was to compare the assessment of medical students communication skills made by their academic teachers, with the assessment made by their role-playing 'patients'. It was a cross-sectional study, conducted at the Department of General Practice, University of Sydney, Australia, and consisted of 519 undergraduate medical students. Teachers rated students' communication skills using ten specific criteria, each marked on a five-point Likert scale. Teachers then rated students' overall performance using a 10-point scale. Patients rated students' overall performance on the same 10-point Likert scale. Only two of the 10 criteria, as rated by the academic teachers, correlated with the role-playing patients' overall score, and all 10 criteria accounted for only 10.1% of the variance in that score. The academic assessors' overall score accounted for only 9.7% of the variance of the patients' overall score. The communications skills emphasized by academic teachers do not reflect the skills considered to be important by role-playing patients. PMID- 9743807 TI - Techniques for rapid quantitative assessment of activity levels in small-group tutorials. AB - Two techniques for the rapid quantitative analysis of student participation in small-group teaching were investigated. In the first approach an observer, who also acted as a 'critical friend', recorded the length of individual contributions using a computer keyboard as a simple timing device. In the second approach, small-group sessions were recorded with a portable stereophonic audiotape recorder. The teacher was recorded on one channel, all students on the other. A computer program produced automated analysis of these small group interactions by computing relative amount of speech on each channel. Simple analysis produced automatically by the programs revealed the overall style of the tutorial--variably 'mini-lectures' by teachers with very little participation by the student body, rapid 'question and answer' sessions with about equal teacher/student body involvement or 'mini-presentations' by students with the teacher offering sparse comments in the manner of a facilitator. By presenting results in a graphic format, teachers can be given rapid objective feedback on their teaching style. Coupled with short verbal/non-verbal quizzes at the end of tutorials and information from other assessments, the value of using levels of participation as a measure of the efficiency of such small-group sessions can itself be assessed. PMID- 9743808 TI - A system for maintaining the educational and training standards of junior doctors. AB - The development of junior doctors' competence is complex because the hospital environment in which doctors work places many demands on them. The need for quality education and training and personal development may be in direct conflict with the service commitments required from hospitals. This paper describes the methods by which the Postgraduate Medical. Council of New South Wales, Australia, addresses the needs of junior doctors in the state in order to improve the quality of their education. Key elements of the Council's function include the provision of hospital clinical supervisors who oversee junior doctor education and training, and central involvement in supplying the junior doctor workforce to all state hospitals who must meet defined accreditation standards. This paper also provides data on evaluation of those methods and some educational outcomes. PMID- 9743809 TI - Working patterns and the quality of training of medical house officers: evaluating the effect of the 'new deal'. AB - The 'new deal' on junior doctors' hours of work has led to the widespread introduction of working patterns such as full shifts and partial shifts in the United Kingdom. The impact of these changes on the training of medical staff is unclear. The subjects of the current study were 36 pre-registration medical house officers working shift rotas and on-call rotas at a teaching hospital in the north of England. They were studied over a 12-month period using a self-report questionnaire seeking their views on the quality of their training experience and their satisfaction with the in-service training they received. Nursing staff, consultant and medical student opinion was also sought. Partial-shift and full shift systems led to reduced hours of work when compared to on-call rotas (mean hours: partial shift 65.0; full shift 59.8; on-call 72.7), but they were associated with significantly lower training experience and training satisfaction scores for the house officers than were on-call systems (P < 0.01). Shift systems were unpopular among consultants and medical students but not nursing staff. Despite reducing excessive hours of work, shifts may be detrimental to the training of medical house officers. The further imposition of shift working should be suspended until such time as the impact of new working patterns on the training of medical staff has been determined. PMID- 9743810 TI - Contribution of medical student research to the Medline-indexed publications of a German medical faculty. AB - Medical students in Germany have to write a research thesis to acquire the title of medical doctor. This study evaluates the contribution of student research to the Medline-indexed publications of a German medical faculty. A 1993-1995 Medline publication list, on which medical students among authors should be marked, was sent to medical faculty staff of the University of Wurzburg, Germany (n = 238). Faculty members responded (106, 45%), 66 were working at a clinic, 26 at a clinic associated institute and 14 at a basic science institute. Between 1993 and 1995, 1128 Medline-indexed papers were published by these faculty members, who on average supervised 4.5 medical students (n = 477). Medical students were among the authors of 316 (28%) and were the first authors of 88 papers (7.8%). For 66% of medical students their research resulted in a Medline-indexed publication. Medical student research activity can significantly influence the published output of a medical faculty. PMID- 9743811 TI - Faculty mentoring programme--ways of reducing anonymity. AB - Increasing anonymity and the lack of personal contact between professor and student due to high enrolment is one of the major problems at German universities. Therefore, the Faculty of Medicine of the University of the Saarland started a pilot project in 1995. The intention was to reduce anonymity within the faculty by introducing a faculty mentoring programme (Studientutorium). A group of up to 12 students from different years is allocated a faculty member on a personal basis. The member responds to questions and problems concerning such areas as the curriculum, career planning or doctoral thesis as well as to personal problems. After 2 years participants were interviewed about the programme for the first time and the acceptance turned out to be high. PMID- 9743812 TI - Chondroitin sulfate and joint disease. AB - Chondroitin sulfate is an important and major component of articular cartilage, where it occurs as part of the large proteoglycan, aggrecan. In the early stages of joint disease, both in animal models and in man, there are changes in chondroitin sulfate that affect the chain length and the pattern of sulfation. These changes can be detected by monoclonal antibodies and appear to reflect part of the cellular response by the chondrocytes to damage to the articular cartilage matrix. The specificity of the changes show that the biosynthesis of chondroitin sulfate is under tight cellular control in chondrocytes and suggests that selected patterns of sulphation within chains are expressed to suit different biological functions. PMID- 9743813 TI - Protective effect of exogenous chondroitin 4,6-sulfate in the acute degradation of articular cartilage in the rabbit. AB - The injection of 2.0 mg chymopapain into the adolescent rabbit knee causes severe loss of articular cartilage proteoglycans (PG). Although chondrocytes attempt to restore lost PG, failure to repair ensues. Pure chondroitin 4,6-sulfate (Condrosulf, IBSA Lugano, Switzerland) has been used in clinical studies of human osteoarthritis (OA) as a slow-acting drug for OA (SYSADOA). Using our model of articular cartilage injury, we examined the effects of oral and intramuscular administration of Condrosulf after chymopapain-induced cartilage injury. In this study, animals received an injection of 2.0 mg chymopapain (Chymodiactin, Boots Pharmaceuticals) into the left knee and were sacrificed after 84 days. The contralateral right knee served as a noninjected control. Some animals received oral Condrosulf while others received intramuscular injections of Condrosulf. Serum keratan sulfate (KS) levels were monitored to ensure degradation of the cartilage PG. Those animals not exhibiting at least a 100% increase of serum KS following chymopapain injection were excluded from the study. At sacrifice, cartilage PG contents were markedly reduced in animals receiving an injection of 2.0 mg chymopapain with no further treatment. In contrast, oral administration of Condrosulf beginning 11 days prior to chymopapain injury resulted in significantly higher (P = 0.0036) cartilage PG contents. Intramuscular administration of Condrosulf resulted in higher, but less significantly so (P = 0.0457), cartilage PG contents. These results suggest that daily Condrosulf treatment prior to and continuing after chymopapain injury may have a protective effect on the damaged cartilage, allowing it to continue to re-synthesize matrix PG after the treatment is discontinued. PMID- 9743814 TI - Anti-inflammatory activity of chondroitin sulfate. AB - The pharmacokinetics of chondroitin sulfate (CS, Condrosulf, IBSA, Lugano, Switzerland) were investigated in rats and in healthy volunteers using CS tritiated at the reducing end and CS labeled with 131I or 99mTc respectively. A rapid absorption of orally administered CS is observed in rats and in humans when the drug is dissolved in water. Lower and delayed absorption is observed when CS is administered in gastroresistant capsules. The absolute bio-availability is 15 and 12% for rats and humans respectively. The CS shows a tropism for cartilagineous tissues in rats and for knee tissues in humans as demonstrated by scintigraphic analysis with 99mTc-CS. Monomers, oligo and polysaccharides produced by enzymatic hydrolysis of CS appear in the blood and tissues together with native CS. The effects of partially depolymerized (m.m. 3 to 15 kD) and desulfated fractions on human leukocytes were investigated. CS and its fractions inhibit the directional chemotaxis induced by zymosan-activated serum, are able to decrease the phagocytosis and the release of lysozyme induced by zymosan and to protect the plasma membrane from oxygen reactive species. In rats the oral administration of CS significantly decreases granuloma formation due to sponge implants and cell migration and lysosomal enzyme release in carrageenan pleurisy. Compared with nonsteroidal anti-inflammatory drugs (indomethacin, ibuprofen), CS appears to be more effective on cellular events of inflammation than on edema formation. It is noteworthy that CS is devoid of dangerous effects on the stomach, platelets and kidneys. In synovial fluid of patients requiring joint aspiration, treated orally for 10 days with CS (800 mg/day) the hyaluronate concentration and the intrinsic viscosity significantly increased, while collagenolytic activity, phospholipase A2 and N-acetylglucosaminidase (NAG) decreased. These results give an insight into the mechanism of the anti inflammatory and chondroprotective actions demonstrated by this drug in a number of clinical trials in patients with osteoarthritis. PMID- 9743815 TI - Practice guidelines in the management of osteoarthritis. AB - Osteoarthritis (OA) is the most common articular rheumatic disease. Practice guidelines have been developed to assist the practitioner in the care of patients with hip and knee OA. The guidelines divide the treatment strategy into nonpharmacologic and pharmacologic modalities. The nonpharmacologic or nonmedicinal approaches include patient education, psychosocial interventions, physical and surgical measures. Each of these are as important as the pharmacologic or medicinal measures. Medicinal measures can be subdivided into symptomatic therapy and disease modifying therapy. Even though all present therapies are aimed at symptoms, experimental therapies are being developed to alter the disease process. PMID- 9743816 TI - Efficacy and tolerability of chondroitin sulfate 1200 mg/day vs chondroitin sulfate 3 x 400 mg/day vs placebo. AB - This multicenter randomized, double-blind, controlled study was performed to compare the efficacy and tolerability of chondroitin sulfate (CS, Condrosulf, IBSA, Lugano, CH) 1200 mg/day oral gel vs CS 3 x 400 mg/day capsules vs placebo, in patients with mono or bilateral knee osteoarthritis (Kellgren and Lawrence radiographic score grade I to III). A total of 127 patients, 40 of whom were treated with CS 1200 mg/day, 43 with CS 3 x 400 mg/day and 44 with placebo, were included in the statistical analysis of this 3-month treatment study. In the CS groups, Lequesne's Index and spontaneous joint pain (VAS) showed a significant reduction of clinical symptoms (P < 0.01 for both parameters), while only a slight reduction was observed in the placebo group (P = ns for Lequesne's Index and P < 0.05 for VAS). The physician's and patient's overall efficacy assessments were significantly in favour of the CS groups (P < 0.01). The treatment carried out with the three formulations was very well tolerated. In conclusion, these results indicate that CS favours the improvement of the subjective symptoms, improving the joint mobility. An additional consideration is that the efficacy of 1200 mg CS as a single daily dose does not differ from that of 3 x 400 mg daily doses of CS for all the clinical parameters taken into consideration. PMID- 9743818 TI - Chondroitin sulfate: S/DMOAD (structure/disease modifying anti-osteoarthritis drug) in the treatment of finger joint OA. AB - A total of 119 patients were included in a randomized, double-blind, placebo controlled trial in order to assess the S/DMOAD properties in OA of chondroitin sulfate (CS 4&6, 3 x 400 mg/day, Condrosulf IBSA, Lugano, CH). Posteranterior roentgenographies of the interphalangeal (IP) joints were carried out at the start of the study and at yearly intervals. This enabled the investigators to document the radiological progression of the anatomical lesions in the pathological finger joints over a 3-year period. It was shown that the progression of OA in the IP finger joints in an individual can be determined by the evolution of his finger joints through previously described anatomical phases: 'N' (not affected), 'S' (classical OA), 'J' (loss of joint space), 'E' (erosive OA) and 'R' (remodeled joint). Structure/disease-modifying anti-OA drug (S/DMOAD) properties were searched for by assaying the number of patients developing OA in previously normal IP joints ('N' > 'S'), or progressing through the described anatomical phases of the disease ('S' > 'J', 'S' > 'E', 'J' > 'E', 'S' > 'R', 'J' > 'R', 'E' > 'R'). In the CS 4&6 group we observed a significant decrease in the number of patients with new 'erosive' OA finger joints. This result is particularly important since OA of the finger joints becomes a clinical problem (pain, functional loss) when 'S' joints progress to 'J' and especially 'E' phases. During and after these 'E' phases, joints will remodel and show the nodular deformities characteristic of Heberden's and Bouchard's nodes. Treated patients were protected against erosive evolution. PMID- 9743817 TI - Efficacy and tolerability of oral chondroitin sulfate as a symptomatic slow acting drug for osteoarthritis (SYSADOA) in the treatment of knee osteoarthritis. AB - Patients with osteoarthritis (OA) of the knee were treated with chondroitin sulfate (CS, Condrosulf, IBSA, Lugano, CH) in a randomized, double-blind, placebo controlled study, performed in two centres. The efficacy and tolerability of oral CS capsules 2 x 400 mg/day vs placebo was assessed in a 6-month study period. Patients with idiopathic or clinically symptomatic knee OA, with Kellgren and Lawrence radiological scores I-III, were included in this trial. Clinical controls were performed at months 0, 1, 3 and 6. Eighty patients completed the 6 month treatment period. Lequesne's Index and spontaneous joint pain (VAS) decreased constantly in the CS group; on the contrary, slight variations of the scores were reported in the placebo group. The walking time, defined as the minimum time to perform a 20-meter walk, showed a statistically significant constant reduction only in the CS group. ANOVA with repeated measures showed a statistically significant difference in favor of the CS group for these three parameters. During the study, patients belonging to the placebo group reported a higher paracetamol consumption, but this consumption was not statistically different between the two treatment groups. Efficacy judgements were significant in favor of the CS group. Both treatments were very well tolerated. All these results strongly suggest that chondroitin sulfate acts as a symptomatic slow acting drug in knee OA. PMID- 9743819 TI - Effects of oral chondroitin sulfate on the progression of knee osteoarthritis: a pilot study. AB - The aim of this study was to assess the clinical, radiological and biological efficacy and tolerability of the SYSADOA, chondroitin 4- and 6-sulfate (CS, Condrosulf, IBSA, Lugano, Switzerland), in patients suffering from knee osteoarthritis. This was a 1-year, randomized, double-blind, controlled pilot study which included 42 patients of both sexes, aged 35-78 years with symptomatic knee OA. Patients were treated orally with 800 mg chondroitin sulfate (CS) per day or with a placebo (PBO) administered in identical sachets. The main outcome criteria were the degree of spontaneous joint pain and the overall mobility capacity. Secondary outcome criteria included the actual joint space measurement and the levels of biochemical markers of bone and joint metabolism. This limited study confirmed that chondroitin sulfate was well-tolerated and both significantly reduced pain and increased overall mobility capacity. Treatment with CS was also associated in a limited group of patients with a stabilization of the medial femoro-tibial joint width, measured with a digitized automatic image analyzer, whereas joint space narrowing did occur in placebo-treated patients. In addition, the metabolism of bone and joint assessed by various biochemical markers also stabilized in the CS patients whereas it was still abnormal in the PBO patients. These results confirm that oral chondroitin 4- and 6-sulfate is an effective and safe symptomatic slow-acting drug for the treatment of knee OA. In addition, CS might be able to stabilize the joint space width and to modulate bone and joint metabolism. This is the first preliminary demonstration that a SYSADOA might influence the natural course of OA in humans. PMID- 9743820 TI - Specialist practice--at what cost? PMID- 9743821 TI - Day hospice care--a review of the literature. AB - This paper traces the development and evaluation of day care hospice provision through analysis of the available literature. The CD-ROM was utilized to access and review the Medline, CINAHL and Healthstar databases. In addition, a hand search of Progress in Palliative Care was conducted. The literature describes the provision of a service within a day hospice, and access to day hospice services. Issues regarding the provision of a day hospice as a mechanism to meet consumer needs are explored. The literature reveals little evaluation of evidence-based practice or of the cost-effectiveness of day hospice provision. There is a dearth of research relating to day care, evaluation studies in particular. It is clearly important that future research compares outcomes with other models of service provision. PMID- 9743822 TI - Measuring quality of life for patients with terminal illness: the Missoula-VITAS quality of life index. AB - Quality of life (QOL) is an important outcome measure in caring for terminally ill patients. The Missoula-VITAS Quality of Life index (MVQOLI) has been developed to provide a measure of quality of life that is meaningful to both clinicians and patients. Unique features of the instrument include its focus on the terminal phase of life, the item structure and a scoring system that allows the weighting of each dimension of QOL by the respondent, and the subjective wording of the items that allows respondents to interpret the measured elements according to their own experience. The validity and reliability of the patient reported survey instrument were tested by administering the 25-item questionnaire to 257 patients in 10 community-based hospices. Participants were incurably ill with predicted survival of six months or less. Exclusion criteria included inability to communicate, dementia, or psychological symptoms that might be intensified by completing the index. Reliability and validity of the new index were examined using standard statistical and psychometrical analyses. The MVQOLI demonstrated internal consistency (Cronbach's alpha = 0.77). MVQOLI total scores were correlated with scores on the Multidimensional Quality of Life Scale--Cancer 2 and with patient-reported global QOL ratings. MVQOLI scores did not correlate with observer-rated functional status scores indicating divergent validity. The MVQOLI could be completed by patients of varied educational level, age, functional status, and length of time with a terminal illness. The instrument is designed to contribute to the task of planning care by evaluating patient identified sources of distress, strength and satisfaction, including issues of life closure. This information contributes to crafting highly specific interventions. Further studies are necessary to determine the usefulness of the instrument in measuring outcomes of end-of-life care in nonhospice settings, and for racial and diagnostic groups under-represented in this sample. PMID- 9743823 TI - Palliative care in chronic obstructive airways disease: a needs assessment. AB - The view that palliative care should move beyond cancer is widely endorsed, however, there remains a lack of clarity about the level at which this should occur. In order to target the palliative approach effectively, the value of more detailed and localized needs assessment becomes apparent. This paper provides evidence from a study commissioned by a department of public health, where the focus was the palliative care needs of an individual with chronic obstructive airways disease (COAD). Over a six-month period, 63 individuals in the district were interviewed about their experiences of living with COAD and the services utilized, using a combination of qualitative and quantitative research methods. The findings revealed a poor quality of life, relating to a high degree of social isolation and emotional distress, associated with low physical functioning and disability, and physical symptoms. Current service provision focused on acute exacerbations. Consequently, there is a need to manage the health and social care interface more effectively, with a shift in emphasis from reactive ad hoc provision, which is where the palliative approach to care could be best suited to meet the needs identified. PMID- 9743824 TI - Sedation for intractable distress in the dying--a survey of experts. AB - Terminal sedation is a phrase that has appeared in the palliative care literature in the last few years. There has not been a clear definition proposed for this term, nor has there been any agreement on the frequency with which the technique is used. A postal survey of 61 selected palliative care experts (59 physicians, two nurses) was carried out to examine their response to a proposed definition for 'terminal sedation', to estimate the frequency of this practice and the reasons for its use, to identify the drugs and dosages used, to determine the outcome, and to explore the decision-making process. Opinions on physician assisted suicide and voluntary euthanasia were also sought. Eighty-seven per cent of the experts responded from eight countries, although predominantly from Canada and the United Kingdom. Forty per cent agreed unequivocally with the proposed definition, while 4% disagreed completely. Eighty-nine per cent agreed that 'terminal sedation' is sometimes necessary and 77% reported using it in the last 12 months--over half of these for up to four patients. Reasons for using this method included various physical and psychological symptoms. The most common drugs used were midazolam and methotrimeprazine. Decision making usually involved the patient or family, and varied with respect to the ease with which the decision was made. The use of sedation was perceived to be successful in 90 out of 100 patients recalled. Ninety per cent of respondents did not support legalization of euthanasia. In conclusion, sedating agents are used by palliative care experts as tools for the management of symptoms. The term 'terminal sedation' should be abandoned and replaced with the phrase 'sedation for intractable distress in the dying'. Further research into the management of intractable symptoms and suffering is warranted. PMID- 9743825 TI - Pain in Chinese cancer patients under palliative care. AB - Over a 4-month period, 218 Chinese patients with advanced cancer were admitted to a palliative care unit in Hong Kong. Ninety-five (44%) of them had pain. Of these 95, 70 (mean age 61.7 years) were evaluated with the visual analogue scale (VAS) and the numerical rating scale (NRS). Forty-two per cent of the 70 patients were illiterate. Eighty-three per cent had metastases. Eighty-seven per cent had pain due to the disease process. Forty-nine per cent had more than one pain, 54% had moderate pain and 20% had severe pain. The most common primary tumour was lung cancer. NRS scores strongly correlated with corresponding VAS scores, suggesting that NRS can routinely be used for pain intensity assessment in the Chinese. The study also showed that 64% of these patients with pain had moderate to severe disabilities to the basic activity of daily living, although there was no correlation between the severity of pain and the severity of disability. PMID- 9743826 TI - How do cancer patients who die at home differ from those who die elsewhere? AB - Our objective was to investigate how cancer patients who die at home differ from those who do not. A postbereavement survey of 229 people who registered the death of a random sample of cancer deaths in an inner London health authority was conducted. It was found that a fifth of patients (21%) died in their own home. Overall, 38% were reported to have expressed a preference for place of death, 73% of whom wanted to die at home. Only 58% achieved this. Having special equipment and stating a preference for place of death was associated with an increased likelihood of dying at home; using social and health services for social care was associated with a decreased likelihood of so doing. It was concluded that, as in previous studies, most patients who expressed a preference wanted a home death, but nearly half did not achieve this. Recognition of a preference for home death, providing the motivation to 'stick it out' at home, and adequate community support to provide the practical means to fulfil the preference, appear to be crucial in the achievement of a home death for all who desire it. PMID- 9743828 TI - Training and knowledge of palliative care of junior doctors. PMID- 9743827 TI - The last month of life: continuity, care site and place of death. AB - A hospice ward was opened in 1991 at the Orebro Medical Centre Hospital (OMCH) in Sweden. Shortly afterwards, a research project was started, which aimed to describe different aspects of the final period of life of a group of cancer patients. This exploratory study is part of this project and aims to assess continuity in the site of care for a group of severely ill cancer patients during the final stages of their lives, and their place of death within different cultures of care. This prospective study involved 56 adults with cancer who had been admitted to six specialized departments at OMCH. Demographic and diagnostic data, documentation of when the patients changed from one care form to another, as well as place of death were obtained. The analysis of continuity in terms of care site involved care-oriented cultures (hospice ward, hospital-based home care, primary care-based home care and nursing home) and cure-oriented cultures (acute hospital wards). Considered as a group, the patients spent one-third of their time at home during their final month of life, with or without formal caregivers. For individual patients, however, there were great variations with regard to continuity of care site and care form. A pattern was found for the type of cancer the patients had and where they were during their final month. Ten patients died in their own homes, and of the 46 who died in an institution, approximately the same number died in a care-oriented culture as in a cure oriented culture. PMID- 9743829 TI - Undergraduate medical education in palliative medicine. PMID- 9743830 TI - Methadone: opioid, N-methyl-D-aspartate antagonist or both? PMID- 9743831 TI - Organ donation: the patients' views. PMID- 9743832 TI - Power and autonomy in palliative care: a matter of balance. PMID- 9743833 TI - Autonomy and its implications for palliative care: a northern European perspective. AB - This paper explores the implications for palliative care practitioners of the pre eminence of autonomy as an ethical principle in contemporary health care ethics. It is suggested that some of the consequences of respecting patient autonomy might be unacceptable to carers, particularly when they feel their own autonomy might be compromised or their ethical values threatened. PMID- 9743834 TI - The use of specialist palliative care services by patients with human immunodeficiency virus-related illness in the Yorkshire Deanery of the northern and Yorkshire region. AB - To examine the use of palliative care services by patients affected by human immunodeficiency virus (HIV) in hospices which do not specialize in the care of HIV patients, a tape-recorded, semistructured interview was carried out in 12 hospices in the UK. The interview explored concerns about such provision, as well as actual issues encountered. The study revealed that all 12 hospices accepted referrals for people affected by HIV and had clear working practices on infection control. between 1990 and 1996, 48 individuals affected by HIV had contact with the hospices. The number of referrals was not related to the size of the hospice. Thirty-nine individuals had a total of 655 days of inpatient care (range 1-35 days); mean length of stay 12.7 days. Twenty-four (62%) died during their first admission. Referrals came from disparate sources and this affected the amount and type of specialist HIV support available to the hospice. The paucity of referrals raised concerns in most of the units as to how to maintain skills. Issues about maintaining confidentiality of diagnosis in a multiprofessional team, and after death were highlighted. All units expressed concerns about the impact on fundraising of HIV-related admissions. Overall it was felt that the hospice units were failing to meet the palliative care needs of the majority of people affected by HIV or acquired immunodeficiency syndrome (AIDS) in the region. Possible reasons for this are given. PMID- 9743835 TI - A telephone survey of the provision of palliative day care services. AB - A telephone survey was conducted to gather preliminary data in order to identify the nature of palliative day care provision in the UK. A random sample of 131 day centres was taken from 17 regional locations in the UK. providing a 60% representation out of a total of 215 adult day care facilities. A combination of a structured and semistructured interview schedule was used to collect the data. The results obtained from this preliminary survey provided further information on the nature, range, and types of services that are currently provided by UK palliative day care centres; management and organizational issues; and the nature of common problems and care issues of patients attending day centres. Data from the study provided further information on the current status of day care services. The implications for future evaluative research are discussed, particularly in the area of the impact and cost-effectiveness of day care services and the potential work with regard to different models of service provision. PMID- 9743836 TI - Development of an aromatherapy service at a Cancer Centre. AB - The aromatherapy service at the Cancer Support and Information Centre (CSIC) of this regional Cancer Centre has been continually assessed since its inception in 1993. New methods of assessing complementary therapies, based on the 'therapy-as practised', have been explored. The present study evaluates the service following changes made after an initial pilot. The professional aromatherapist developed an evaluation tool, and formal questionnaires were limited to the Hospital Anxiety and Depression Scale (HADS). HADS was completed before and after a course of six aromatherapy sessions. Of 89 patients referred, 58 patients completed the six sessions. Referrals were made by health professionals working in the Cancer Centre and in the CSIC. The majority of patients were female with breast cancer and were receiving radical oncological treatment. Tension, stress and anxiety/fear were the most common reasons for referral, and this was reflected in high initial HADS scores. There were significant improvements in HADS scores in the 58 patients completing the course (mean anxiety, depression, and combined scores dropped from 8.9 to 6.2 6.1 to 4.0 and 15.0 to 10.2, respectively, P < 0.001). Fifty per cent or more of the sample reported a significant improvement in the eight most commonly assessed symptoms. The therapist was initially cautious about using questionnaires, but she gained confidence in using HADS as an assessment tool. The areas covered by her own evaluation tools were broadly comparable to established instruments such as the EORTC QLQ-C30. We conclude that aromatherapy massage has a role in reducing psychological distress, and improving symptom control in cancer patients. Further service evaluation is needed to promote appropriate referral and effective planning of treatment, and to justify cost. Given the multifaceted nature of complementary therapies, the need to develop new research methodologies is acknowledged. PMID- 9743837 TI - An annotated bibliography of the publications of Cicely Saunders--1: 1958-67. PMID- 9743838 TI - My father died of dementia.... PMID- 9743839 TI - Formative evaluation and its relevance to palliative care. PMID- 9743840 TI - Chronic bleeding secondary to an unresectable duodenal adenocarcinoma controlled with sucralfate and famotidine. PMID- 9743841 TI - Risperidone in the management of agitation in HIV dementia. PMID- 9743842 TI - Use of enteral glycopyrrolate in the management of drooling. PMID- 9743843 TI - Thalidomide for distressing night sweats in advanced malignant disease. PMID- 9743844 TI - Thoughts of self-harm in terminally ill patients. PMID- 9743845 TI - Managing gastric stasis. PMID- 9743846 TI - The EMO vaporizer. PMID- 9743847 TI - Amrinone improves right ventricular ejection fraction and oxygen delivery without deterioration of extravascular lung water in canine oleic acid pulmonary injury. AB - Fourteen mongrel dogs were anaesthetized and mechanically ventilated with oxygen. After oleic acid was administered intravenously, amrinone 1 mg/kg was intravenously administered to one group followed by a continuous infusion of 10 micrograms/kg/min for one hour (amrinone group, n = 7). Isovolumetric saline was administered to the control group (n = 7). Amrinone slightly lowered PaO2 but significantly increased oxygen delivery and improved gastric intramucosal pH (pHi) during the first 30 minutes (7.33 +/- 0.13) compared with the control group (7.21 +/- 0.06, P < 0.05). The pHi remained higher in the amrinone group (7.30 +/ 0.15) than in the control group (7.16 +/- 0.06, P < 0.05) after the drug was withdrawn. Extravascular lung water was significantly augmented after oleic acid injection and sustained in all animals for the remainder of the study. PMID- 9743848 TI - Induction of anaesthesia with sevoflurane, preprogrammed propofol infusion or combined sevoflurane/propofol for laryngeal mask insertion: cardiovascular, movement and EEG bispectral index responses. AB - Inhalation induction with sevoflurane was compared with propofol or sevoflurane/propofol in 60 unpremedicated adults. Target concentrations for the three groups (with 60% nitrous oxide) were 3% end-tidal sevoflurane, 12 mg/l propofol and 1.5% sevoflurane/6 mg/l propofol respectively, prior to insertion of a laryngeal mask airway (LMA) at 10 minutes. Induction of anaesthesia was satisfactory in each group, but movement response to LMA insertion was observed in 20 patients (least in the sevoflurane group). Cardiovascular responses were similar except for a lower heart rate in the sevoflurane group. EEG bispectral index suggested a greater depth of anaesthesia in the inhalation induction group. A bispectral index of 60 separated patients responding to LMA insertion from nonresponders (P = 0.006), and had a sensitivity of 68% and specificity 70%. Movement response was not predicted by cardiovascular changes. PMID- 9743849 TI - Lack of efficacy of propofol in the treatment of early postoperative nausea and vomiting. AB - The anti-nauseant efficacy of low-dose propofol was investigated in a blinded, randomized trial. Patients who complained of nausea and/or vomiting following laparoscopic gynaecological surgery and who requested antiemetic were randomly assigned to receive placebo, propofol 3 mg, propofol 9 mg or propofol 27 mg by intravenous injection. Nausea, vomiting and sedation were recorded by a blinded observer for 90 minutes following administration of the test drug, prior to discharge, and 24 hours following surgery. Rescue antiemetic (droperidol 1.0 mg i.v.) was available from 10 minutes after administration of test drug. Propofol failed to reduce nausea scores and did not reduce the incidence of vomiting. Numbers of patients receiving rescue antiemetic were similar in the four treatment groups. In the first 10 minutes following test drug administration, sedation scores were increased by propofol in a dose-related manner. We conclude that, in the dose range studied, propofol is ineffective for the treatment of nausea and vomiting occurring soon after laparoscopic gynaecological surgery. PMID- 9743850 TI - Reinforcing a "low flow" anaesthesia policy with feedback can produce a sustained reduction in isoflurane consumption. AB - A three-month audit of isoflurane consumption at Palmerston North Hospital in 1994 showed an averaged vapour flow rate of approx 85 ml per minute of anaesthesia, equivalent to 1.4% isoflurane at six litres per minute. After purchasing volatile agent analysers, a program encouraging low flow anaesthesia and providing a report of the previous month's consumption rate was started in July 1996. The isoflurane averaged vapour flow rate was tracked over the following twenty-month period and fell by a sustained 65% to range around 30 +/- 5 ml/min, producing savings of approximately NZ$104,000 over this period. PMID- 9743851 TI - Incidence of phrenic nerve block and hypercapnia in patients undergoing carotid endarterectomy under cervical plexus block. AB - Deep cervical plexus blockade blocks the second, third and fourth cervical nerve roots. The phrenic nerve arises from C3, C4, C5 and should therefore be commonly blocked with cervical plexus blockade. The aim of this study was to report the incidence of phrenic nerve block and to assess the effect of this on arterial carbon dioxide tension (PaCO2) in premedicated and sedated patients. Forty patients were studied, blood gases being taken on the day before surgery, immediately before performing the block and then every 20 minutes until the operation was completed. Fluoroscopy was used to determine ipsilateral hemidiaphragmatic dysfunction due to phrenic nerve block. The patients were then divided into two groups of analysis. Group A patients had evidence of phrenic nerve block, Group B patients had no evidence of phrenic nerve block. Fluoroscopy showed that 22 patients (55%) had evidence of phrenic nerve block (Group A), 18 patients showed no change (Group B). PaCO2 levels increased in both groups following premedication, from 41 +/- 5 mmHg (mean +/- SD) to 46 +/- 5 mmHg in Group A, and 41 +/- 4 mmHg in Group B; twenty minutes after cervical plexus block the PaCO2 rose to 49 +/- 6 mmHg in Group A, and 48 +/- 6 mmHg in Group B. These changes were not statistically significantly different when the two groups were compared. PMID- 9743852 TI - Screening tests for predicting difficult intubation. A clinical assessment in Turkish patients. AB - Three methods of predicting difficult intubation were compared prospectively. Mallampati test, Wilson risk-sum and thyromental distance were determined preoperatively and laryngeal views were graded in 500 patients. The sensitivities, specificities, positive and negative predictive values of each test were calculated. The incidence of difficult intubation was found to be 8%. The sensitivities of the Mallampati test, the Wilson risk-sum and the thyromental distance were 43%, 58% and 35% respectively, and the specificities were 93%, 91% and 95% respectively. Significant differences were seen between the sensitivities of the Mallampati test and the Wilson risk-sum (P < 0.001), the Wilson risk-sum and the thyromental distance (P < 0.001), the Mallampati test and the thyromental distance (P < 0.05). Among the different specificities, the only significant difference was observed between the Wilson risk-sum and the thyromental distance (P < 0.05). There were no significant differences between the positive and negative predictive values of the three screening tests (P > 0.05). In conclusion, the Wilson risk-sum was the most sensitive test and the thyromental distance has the highest positive predictive value for difficult intubation. PMID- 9743853 TI - Tracheal intubation through the intubating laryngeal mask airway (LMA-Fastrach) in patients with difficult airways. AB - The intubating laryngeal mask airway was used in 31 adult patients in whom tracheal intubation was known or suspected to be difficult. The intubating laryngeal mask airway was successfully inserted in 30 patients and provided a clinically patent airway. In the remaining one patient it was impossible to insert the device correctly. Tracheal intubation through the device was successful in 28 of 30 patients (93%). These results suggest that the intubating laryngeal mask airway has a potential role for tracheal intubation in adult patients with difficult airways. PMID- 9743854 TI - Effect of nutritional status on vecuronium induced neuromuscular blockade. AB - Based on simple clinical and biochemical parameters of nutritional status, seventy adult patients scheduled for elective surgery under general anaesthesia were categorized as having normal nutrition, mild, moderate or severe malnutrition or obesity. Under anaesthesia, evoked responses on train-of-four nerve stimulation were recorded every 15 seconds on a mechanomyograph. Vecuronium 0.1 mg.kg-1 was used to achieve neuromuscular blockade. Compared with patients having normal nutrition, the time to onset of action was significantly prolonged in the moderate and severely malnourished groups; the time to no response on train-of-four stimulation was delayed only in severely malnourished groups (P < 0.001). The duration of action of the initial dose was shorter in the moderate and severely malnourished groups. The obese group had an earlier onset of action and a longer duration of action compared with patients of normal nutrition (P < 0.001). No significant difference in recovery time to a train-of-four ratio of 0.70 was observed between the malnourished and patients with normal nutrition. Malnutrition has a marked effect on vecuronium-induced neuromuscular blockade. PMID- 9743855 TI - Problems associated with nursing staff shortage: an analysis of the first 3600 incident reports submitted to the Australian Incident Monitoring Study (AIMS ICU). AB - Although many studies have attempted to define appropriate nursing staff levels, allocation and patient dependency, minimal data is available on the effect of nursing staff shortage (NSS) on quality of care provided in intensive care. This study aimed to identify incidents associated with staff shortage as reported to the Australian Incident Monitoring Study-ICU (AIMS-ICU) project and to assess their estimated effect on patient outcome. A search of narrative keywords and contributing factors identified 89 nursing staff shortage incidents (NSS INCIDENTS) and 373 incidents involving nursing staff shortage contributing factors (NSS-CF). NSS resulted from inappropriate rostering for current patient load (81%) and inability to respond to increased unit activity (19%). Most frequent associated incidents included problems with: drug administration/documentation (47), patient supervision (20), set-up of ventilators/equipment (16), and accidental extubation (14). Undesirable patient outcomes included: major physiological change (22%), patient/relative dissatisfaction (12%), and physical injury (3%). This study suggests that inadequate staffing results in incidents and compromised patient safety. PMID- 9743856 TI - The third five-year survey of fellows (by examination) of the Faculty of Intensive Care, Australian and New Zealand College of Anaesthetists. AB - A questionnaire was sent to 126 Fellows who had passed the Fellowship Examination in Intensive Care up to and including the examination of October, 1995. The major objectives were to assess the continuing involvement of Fellows in Intensive Care and obtain feedback on training and the examinations. Only six Fellows failed to respond. Ninety-six per cent of responders had some involvement in Intensive Care and 89% had a current formal Intensive Care appointment. The median percentage of the week spent in the Intensive Care was high. Forty-seven per cent were practising some anaesthesia. Although there was considerable individual variation, the Fellows had not changed their median amount of Intensive Care practice over time. The responders provided feedback on their work patterns in the public and private systems, and their training and examinations. Overall, the training/examination system appears to satisfy Fellows although some fine tuning is required. PMID- 9743857 TI - Labour ward midwifery staff epidural knowledge and practice. AB - A survey was conducted amongst labour ward midwives at our hospital to evaluate education, knowledge and attitudes toward the management of epidural analgesia in labour. Sixty of 80 distributed forms were returned, giving a 75% response rate. Forty-two per cent of respondents had more than ten years' practice experience. Only 51% achieved a predetermined pass score for knowledge about epidural analgesia. Though most had received formal education about epidural analgesia, 35% felt postgraduate education was insufficient. A majority of midwives supported epidural analgesia on demand (78%), during established labour (74%), or for women at increased risk of caesarean section (82%). Midwives with lower knowledge levels were more likely to recommend epidural analgesia early in labour to multiparous women (P = 0.001) and to women with either a small or a large baby (P = 0.06). The majority of midwives (93%, 70% and 65%) would "almost always" top up women with cardiac or medical diseases, multiple pregnancy or hypertensive disease, respectively. A clear requirement for ongoing education, with input from the anaesthetic department, was identified, irrespective of personal experience. Practice patterns are discussed and recommendations made with respect to improvement of epidural analgesia management and continuing education. PMID- 9743858 TI - Acute hyponatraemia secondary to cerebral salt wasting syndrome in a patient with tuberculous meningitis. AB - A 30-year-old HIV-positive man presented with acute hydrocephalus secondary to tuberculous meningitis, for which an external ventricular drain was inserted. He developed marked natriuresis in the postoperative period, which resulted in acute hyponatraemia (131 to 122 mmol/l) and a contraction of his intravascular volume. A diagnosis of cerebral salt wasting syndrome was made, and he responded to sodium and fluid loading. This case highlights the differentiation of cerebral salt wasting syndrome from the more commonly occurring syndrome of inappropriate anti-diuretic hormone secretion as the aetiology of the hyponatraemia. PMID- 9743859 TI - Paraspinal abscess associated with epidural in labour. AB - A 24-year-old presented in labour requesting an epidural. She had been diagnosed as having "pelvic arthropathy" at 37 weeks. After an uneventful epidural and instrumental delivery, she discharged herself home. She re-presented 19 days later with left hip pain and abnormal neurological signs in the lower limbs. On MRI, there was a large paraspinal abscess with an epidural granulation mass compressing her spinal cord. She had an urgent surgical decompression. In hindsight, it is likely that the paraspinal mass was present at the time of epidural insertion. The discussion highlights that complications are sometimes not what they seem. PMID- 9743860 TI - Explosion and fire in the expiratory limb of a Fisher and Paykel "three in one" respiratory care system. AB - We report an incident involving a Fisher and Paykel "There in One Respiratory Care System" (Fisher and Paykel Healthcare, Auckland, New Zealand). This is a new ventilator circuit designed to be adaptable to the needs of ventilator, intubated CPAP, and mask therapies. In this case the patient had received eight hours of CPAP therapy overnight, during which time the "Three in One" circuit had been broken down to the CPAP configuration. The expiratory limb heater element, normally disconnected, was inadvertently left connected to the heater base. Under the extreme conditions of heating under "no gas flow" mode, the heater element malfunctioned. As a result when the circuit was reconfigured to provide pressure supported ventilation, in the high oxygen environment of the expiratory limb (FiO2 0.5), ignition and combustion of the respiratory circuit occurred. The case is reported because of the potentially serious consequences and because the incident prompted the manufacturer to redesign and change componentry in parts of the circuit implicated. PMID- 9743861 TI - Vale epidural anaesthesia? PMID- 9743862 TI - Fractured fine gauge, narrow, atraumatic (pencil point), spinal needles. PMID- 9743863 TI - Inhalational induction with sevoflurane in central airway obstruction. PMID- 9743864 TI - Balanced analgesia for the management of pain associated with multiple fractured ribs in an opioid addict. PMID- 9743865 TI - Ropivacaine 0.5% and bupivacaine 0.5% epidural blockade for lower limb orthopaedic surgery. PMID- 9743866 TI - The case for the use of verapamil in alarming Chironex stings. PMID- 9743867 TI - Related measurements do not imply agreement. PMID- 9743868 TI - Human tumour xenografts in nude mice: chemotherapy trials with titanocene dichloride in different dosages. AB - PURPOSE: In this study new cytostatic therapies with titanocene dichloride in different dosages for the treatment of ovarian cancer are analyzed on human tumour xenografts in nude mice. The aim was to compare the effects of different dosages of titanocene dichloride on the growth of human tumour xenografts and nude mice body weight. METHODS: Biopsy material from one human ovarian carcinoma was expanded and transplanted into 52 nude mice. The treatment protocol included one experiment that consisted of the following six treatment groups: titanocene dichloride 3 x 10 mg/kg, titanocene dichloride 3 x 20 mg/kg, titanocene dichloride 3 x 30 mg/kg, titanocene dichloride 1 x 30 mg/kg, titanocene dichloride 1 x 40 mg/kg and a control group treated with 0.9% saline. Treatment groups were evaluated in terms of average daily increase in tumour volume and average daily body weight increase on nude mice. The slope factors alpha and beta of the body weight and tumour volume changes were calculated. RESULTS: Titanocene dichloride in the dosage of 3 x 30 mg/kg and 3 x 20 mg/kg brought about a significant reduction in tumour volume (p < 0.05) compared to the control group and to the treatment group under medication with titanocene dichloride 1 x 30 mg/kg. There were no significant changes in the body weight of nude mice. CONCLUSION: We found titanocene dichloride to be effective in the reduction of tumour volume increase in nude mice. Titanocene dichloride could be an active chemotherapeutic drug in women with ovarian carcinoma not responding to standard therapies. PMID- 9743869 TI - C-reactive protein levels at the onset of labour and at day 3 post-partum in normal pregnancy. AB - OBJECTIVES: To record maternal serum C-reactive protein levels during normal onset of labour and normal puerperium and to evaluate if inflammation or infection could be predicted during these two periods when serum C-reactive protein is increased. METHODS: Eighty-five pregnant women were enrolled in a longitudinal prospective study and had a blood sample to assess serum C-reactive protein levels on admission to the labour ward for normal onset of labour and at day three post-partum. Inclusion criteria were no previous history, a normal single pregnancy, normal vaginal delivery and an uneventful post-partum course. Twelve non-pregnant women of the same age constitued a control group. An automatic Behring Nephelometer was used to measure serum C-reactive protein concentrations. The Student's t-test (significance p < 0.05) was used for statistical analysis. FINDINGS: C-reactive protein was significantly increased during the onset of labour (4.10 +/- 2.79 mg/L) and reached very high levels during the post-partum period (24.07 +/- 18.28 mg/L) compared to the standard normal serum C-reactive protein level in a population of non-pregnant women of the same age (2.39 +/- 0.07 mg/L). INTERPRETATION: Increased serum C-reactive protein has been reported to be a marker for subclinical infection during pregnancy in various situations including premature labour and premature rupture of membranes and for complications occurring during puerperium such as thrombophlebitis, thromboembolism or endometritis. This interpretation depends on which upper limit is considered as abnormal. Because serum C-reactive protein was raised during the onset of labour, values of less than 10 mg/L could not be considered as a marker for infection during this period. Elevated serum concentrations of estrogen, progestogen and prostaglandins during labour might be one explanation for those physiological changes. Normal vaginal delivery could be compared to a surgical procedure and tissue injury consecutive to vaginal birth as reflected by a dramatic increase in C-reactive protein. More studies using nephelometry are needed to determine normal and upper values of C-reactive protein during pregnancy. PMID- 9743870 TI - Ovarian blood flow before and after conservative laparoscopic treatment for endometrioma. AB - To evaluate the vascular changes in ovaries affected by endometriomas 28 women with ovarian endometriosis underwent transvaginal ultrasound with color flow imaging and blood flow analysis of the ovarian artery before and after laparoscopic conservative treatment of the ovarian cyst. Mean pulsatility index (P.I.) and resistance index (R.I.) of the ovarian artery on the side affected by endometrioma were compared using Student's t-test. Mean P.I. after laparoscopy (1.59) was significantly lower (p = 0.001) than before surgical intervention (2.17). Analogously the mean R.I. was significantly different (p = 0.001) when compared before (0.81) and after (0.73) laparoscopy. Color Doppler velocimetry may add greater understanding of the ovarian hemodynamic changes that occur after conservative surgery on the ovary. PMID- 9743871 TI - Strongly suspected microangiopathic hemolytic anemia associated with radiotherapy in the treatment of advanced cervical cancer. PMID- 9743872 TI - Compliance with hormone replacement therapy in postmenopausal women. A comparative study. AB - OBJECTIVE: To assess compliance with hormone replacement therapy in postmenopausal women. METHOD: Two groups were compared prospectively: 100 women who sought treatment for menopausal symptoms, and 82 women who had undergone a total abdominal hysterectomy with bilateral salpingo-oophorectomy and were using estrogen replacement therapy. RESULTS: Compliance rates after 6 months were 81.0% and 84.1% in the two groups, respectively, and after 12 months, 73.0% and 80.5%. CONCLUSIONS: The high rates are attributed to our investment in patient education of the benefits of treatment and repeated and close follow-up. PMID- 9743874 TI - Evaluation of fetal movements as an early labour admission test in low-risk pregnancies. AB - Fetal movements were quantified in 182 low-risk women in early labour using the Hewlett-Packard M1350A (Boblingen, Germany) fetal heart rate monitor. There were no statistically significant differences in adverse intrapartum or neonatal outcomes detected by the fetal heart rate pattern or fetal movement profile. This study confirms the feasibility of obtaining, a measure of fetal movement in early labour but does not support its use as an admission test in low-risk pregnancies. PMID- 9743873 TI - Comparison of somatomedin-C (SMC/IGF-I), human placental lactogen and Doppler velocimetry between appropriate and small-for-gestational-age pregnancies. AB - Thirty-two pregnant women with small-for-gestational-age (SGA) fetuses and 45 pregnant women with appropriate-for-gestional-age (AGA) fetuses (controls) were recruited after the 32nd week of gestation. Blood samples were collected for estimation of somatomedin-C (SMC/IGF-I) and hPL in the maternal serum and in the umbilical cord serum. The systolic/diastolic (S/D) ratio of the umbilical artery was also recorded. The results showed somatomedin-C and hPL levels in the maternal serum and in the umbilical cord to be significantly decreased and the Doppler S/D ratio to be significantly increased in the SGA group. In this group, using the multivariable regression analysis, we found significant correlations between maternal hPL, somatomedin-C, Doppler S/D ratio and birth weight. PMID- 9743875 TI - Agreement between hysterosalpingography and laparoscopic chromopertubation in assessment of tubal patency. AB - BACKGROUND: To assess the agreement between tubal patency assessed by laparoscopy with chromopertubation and by hysterosalpingography using contrast media. SETTING: University Medical School. PATIENTS: 314 consecutive women subjected to laparoscopy and hysterosalpingography for an infertility study. DESIGN: Prospective study. METHODS: Chromopertubation using Methylen blue dye, performed on days 20-24. Hysterosalpingography performed on days 7-10 with water soluble contrast. MAIN OUTCOME MEASURE: Kappa coefficient calculation. RESULTS: Kappa coefficient ranged from 0.40 to 0.36, depending on the categories analyzed, corresponding to a fair agreement. CONCLUSION: The diagnosis of tubal factor requires that both tubal patency tests (Hysterosalpingography and laparoscopy) show an abnormal patency. When one of the aforementioned tests is normal, performing the second one has little clinical advantage. However, it is suggested that when there is a discordant patency the pregnancy rates could be somewhat reduced. PMID- 9743876 TI - Pregnancy and delivery in a group of Israeli teenagers. A case-controlled study. AB - OBJECTIVE: To assess characteristics of an Israeli group of nulliparous teenagers and to compare selected variables of their course and outcome of pregnancy with controls. METHODS: Hospital records of 46 consecutive nulliparous teenagers younger than 17.5 years who delivered during a ten-year period and 84 matched adult controls were reviewed. RESULTS: The majority of the teenagers were older than 15 years, married and most were born in Israel or in the former Soviet Union with no obvious socio-economical deprivation. The rate of prenatal follow-up, hypertensive disorders, type of analgesia during labor and mode of delivery were similar in teenagers and controls. A statistically non-significant higher rate of anemia (hemoglobin, 10 gr%), preterm delivery and low birth weight were observed in teenagers. Only the rate of induction of labor and the rate of a hemoglobin level higher than 12 gr% were significantly lower in teenagers. CONCLUSIONS: The course and outcome of pregnancy were in most respects similar in this group of nulliparous teenagers and matched adult controls. PMID- 9743877 TI - Gynecologic cancer and surgical infectious morbidity. AB - The purpose of this study was to evaluate retrospectively the surgical infectious morbidity in gynecologic cancer. We examined 1,180 gynecologic oncology patients: 608 women had carcinoma of the endometrium, 510 cancer of the cervix, 48 ovarian cancer and 14 vulvar cancer. Thirty-five (6%), 92 (18%), 7 (15%) and 2 (14%) were complicated by infection in carcinoma of the endometrium, cancer of the cervix, ovarian cancer and vulvar cancer, respectively. Our conclusion is that the highest surgical infectious morbidity occurs in patients with cervical cancer and the lowest in patients with carcinoma of the endometrium. PMID- 9743878 TI - Effect of age and parity on primary caesarean section rates. AB - OBJECTIVE: To study the effect of maternal age and parity on the rates of primary caesarean section. METHOD: We reviewed all patients who delivered at the Princess Badeea Teaching Hospital between 1 January, 1995 and 26 November, 1995. RESULTS: There were 8,732 deliveries included in this study. The primary caesarean section rates in primiparous women less than 25, 25 to 34 and over 34 years of age were 6.1%, 11.1% and 22.2%, respectively. A similarly dramatic rise with advancing maternal age was seen in multiparous women with rates of 3.1%, 6.4% and 9.5%, respectively, in the three age groups. A strong association between maternal age and primary caesarean section exists (p < 0.05). Caesarean section rates in the primiparous women were higher in all age groups when compared with multiparous women (p < 0.0001). CONCLUSIONS: Increasing maternal age and parity are factors strongly associated with increased primary caesarean section rates. PMID- 9743879 TI - Onset of erythema nodosum during pregnancy: a case report. PMID- 9743880 TI - Effects of gestation age and of birth weight in the concentration of carnitine in the umbilical plasma. AB - To investigate the factors which affect the concentrations of the total, the free, and the acylcarnitine in neonates, blood was taken from the umbilical cord of 49 newborn infants ranging in gestation age (g. a.) from 32-40 weeks (mean g. a.: 36.8 +/- 2.6 weeks) and in birth weight (b. w.) from 1300 gr.-4300 gr. (mean b. w.: 2299 +/- 457 gr.). The carnitine and its fractions were studied in plasma. Twenty-eight of the neonates studied were premature (g. a. < or = 37 weeks) and 21 were full-term (g. a. > 37 weeks). The concentration of the total, free, and acylcarnitine in premature neonates was 28.0 +/- 2.3 mumol/L, 15.9 +/- 1.3 mumol/L, and 12.0 +/- 1.3 mumol/L, respectively. For the full-term neonates the respective concentrations were: 25.2 +/- 2.2 mumol/L, 14.6 +/- 1.5 mumol/L, and 10.7 +/- 1.5 mumol/L. These differences in concentrations between premature and full-term infants were statistically significant. For the total number of neonates studied the concentration of total, free, and of acylcarnitine was 26.8 +/- 2.6 mumol/L, 15.3 +/- 1.9 mumol/L, 11.5 +/- 1.5 mumol/L respectively. The calculation of the correlation coefficients for the total number of neonates showed the existence of a statistically significant negative correlation between the total, free and acetyl carnitine in terms of gestation age and birth weight. The comparative analysis of the correlation coefficients showed greater coefficient values between the total and the acylcarnitine in terms of birth weight. The latter finding, combined with the low rate of acylcarnitine decline, are indirect indications that the fetus uses carnitine as a source of energy, which affects the levels of total and acylcarnitine in the plasma. PMID- 9743881 TI - A randomized trial of intracervical prostaglandin gel and intravenous oxytocin in prelabor rupture of membranes with unripe cervix at term. AB - In order to compare the efficacy of immediate intravenous oxytocin administration and intracervical prostaglandin E2 gel application in premature rupture of membranes with unfavorable cervices at term, 45 term pregnant patients with premature rupture of membranes were randomized into two groups. Twenty women received immediate intravenous oxytocin after cleansing enema while the rest were treated with intracervical prostaglandin E2 gel. Means of maternal age, gestational age, Bishop score at admission and the rates of nulliparity did not show any significant differences between the two groups (p > 0.05). The mean rupture to delivery time was 12.6 +/- 4.4 hours in the oxytocin group and 16.5 +/ 4.5 hours in the prostaglandin group (p < 0.01). Mean birth weights and Apgar scores were insignificant. Cesarean section rates were 24% in the oxytocin group and 5% in the other (p < 0.05). No infectious morbidity was seen in any case. In conclusion, although delivery is delayed with the intracervical prostaglandin approach, cesarean section rate is lowered without an increase in infectious morbidity. PMID- 9743883 TI - Dystocia: is it a major indication for caesarean section? AB - OBJECTIVE: To find out the indications for caesarean sections, the contribution of "dystocia" to the overall caesarean section rates, and to find ways to reduce dystocia-induced caesarean sections. METHOD: This was a retrospective study where all caesarean sections performed in 1995 at the Princess Badeea Teaching Hospital in North Jordan (the main teaching and referral hospital in the area) were reviewed. RESULTS: The caesarean section rate for 1995 was 8.4%. Dystocia was the main indication in 13.4% of all caesarean sections in that year. In 80.2% of patients who delivered because of dystocia labour started spontaneously. Thus if we advocate active management of labour, especially in nulliparous women who start labour spontaneously due to dystocia, we may reduce caesarean section and many repeat caesarean sections could be avoided. CONCLUSIONS: Applying a policy of active management of labour in nulliparous women may be the most useful approach to reduce caesarean section rates in modern obstetric practice. PMID- 9743882 TI - Laparoscopic treatment of interstitial pregnancy using the harmonic scalpel. AB - OBJECTIVE: To evaluate a laparoscopic technique of cornual resection using the harmonic scalpel. METHODS: Four patients with unruptured interstitial pregnancies were treated laparoscopically using the harmonic scalpel at Ioannina University Hospital. RESULTS: There were no failures in the technique in any of our patients. CONCLUSION: Our study suggests that interstitial pregnancies of a maximum gestational age of 7-8 weeks and sac diameter less than 4-5 cm may be treated laparoscopically. PMID- 9743884 TI - Conservative treatment of multiple pregnancies after delivery and a fetal miscarriage: two case reports. AB - Due to the increased availability of infertility treatment, multiple pregnancies, with various resulting complications, have become more common. Two cases of triplet pregnancies with delayed delivery--interval ranged from 6 to 56 days--are reported. The first woman in the 23rd week of a triplet pregnancy came to the hospital because of premature rupture of membranes of one amniotic sac and had a miscarriage of one of the fetuses the same day. The second woman in the 26th week of a triplet pregnancy also came to the hospital because of bleeding and contractions and had vaginal delivery of the first triplet a few hours later. After confirming that the remaining two fetuses were in good condition, both patients were kept under observation with only antibiotic therapy for the first one and antibiotic and tocolysis for the second. Fifty-six days following admission to our hospital the first woman gave birth to twins while in her 32nd week. The second woman gave birth six days following admission (in her 27th week). The successful outcome of these cases demonstrates that non-intervening conservative methods could be a feasible alternative to invasive intervention. We hope that our cases will encourage more physicians to try out and report non intervening methods so that enough information can be gathered to help make correct management decisions in the future. PMID- 9743885 TI - Can we reduce repeat caesarean delivery at the Princess Badeea Teaching Hospital in north Jordan? AB - OBJECTIVE: Our aim was to describe the indications of repeat caesarean delivery and to determine modifiable practice patterns that might lead to fewer repeat caesarean deliveries. METHOD: Hospital records of all women with previous caesarean sections who delivered between 15 April, 1994-31 December, 1994 at the Princess Badeea Teaching Hospital in North Jordan were reviewed. Three groups were identified: 1) elective repeat caesarean 2) vaginal birth after caesarean 3) failed vaginal birth after caesarean. RESULTS: In this study there were 388 patients. Of these, 208 had a repeat caesarean delivery for the following reasons: failed vaginal birth after caesarean (39, 10.1%) and repeat elective caesarean section (169, 43.5%). The remaining (180, 46.4%) patients had a vaginal birth after caesarean. CONCLUSIONS: Our vaginal birth rate after one previous caesarean section was 82.2%. If this rate can be maintained in patients with 2 or 3 previous caesarean deliveries, we can reduce repeat caesarean rates by at least 14% by allowing more patients with 2 or even 3 previous caesarean deliveries to have a trial of labour under appropriate conditions and also proper management of dystocia. PMID- 9743886 TI - Cytokine levels in seminal plasma. AB - We have studied IL-2, SIL-2R, and sCD4 concentrations in the seminal plasma of 20 healthy men (controls), 20 oligozoospermics and 20 azoospermics by the "sandwich" enzyme immunoassay technique. We have also measured IL-2, SIL-2R and sCD4 in the two fractions of the split ejaculate of 10 healthy men. Our results show that the mean IL-2 and sCD4 concentrations in the seminal plasma were significantly higher compared to the levels in the serum of normal men; furthermore, there were no significant differences of SIL-2R and sCD4 between the group of normal men and the groups of men with oligozoospermia or azoospermia. On the contrary, significantly higher levels of IL-2 and sCD4 were found in the first compared to the second fraction of the split ejaculates. Our results support the view that the estimation of IL-2 in seminal plasma has value concerning the quality of semen. On the contrary, the estimation of SIL-2R and sCD4 in seminal plasma are valueless concerning normal or abnormal semen. Finally, the prostate seems to be the main site or origin of IL-2 and sCD4. PMID- 9743887 TI - A case of Meigs' syndrome with a gigantic granulosa ovarian tumor. PMID- 9743888 TI - Short time effect of Chemiron (a combination iron preparation), single iron, and different magnesium salts on plasma. Magnesium concentration during early pregnancy in Nigerian women. A preliminary report. AB - Maternal magnesium requirements increase during pregnancy because of the synthesis of new tissue--both fetal and maternal. Magnesium takes part in almost 300 enzymatic reactions in the human body and regulates membrane permeability and protein bio-synthesis by promoting initiation and dissociation factors. The absorption velocity of magnesium differs from one tissue to another in animal experiments. It is highest in the liver, kidney, heart and is low in skeletal muscle, the brain and erythrocytes. It obeys and follows the Michaelis-Menten Kinetic law. 15 mmol of magnesium is consumed daily depending on the types of food takenin. The main sources of magnesium are vegetables and meats. Many Nigerian women are not able to afford enough of these. The amount of magnesium reabsorbed depends on the magnesium intake and not on magnesium needed which is about 10-40% of the intake. In this study, we examined the short-term effect of magnesium asphat HCL (614.18 mgMG), magnesium diasporal (magnesium citrate 610 mg + magnesium laevalitat 30 mg = 100 mg magnesium = 8.2 mval), ferrous gluconate (300 mg) plus folic acid and chemiron, a new combination hematinic agent (ferrous fumarate 300 mg, folic acid 5 mg, vitamin B12 10 mg, vitamin C 25 mg, magnesium sulfate 0.3 mg and zinc sulfate 0.3 mg) on plasma magnesium concentration during early pregnancy in Nigerian women. Significant increases of plasma magnesium concentrations were found in these groups (magnesium asphat HCL, 0.83 +/- 0.12 to 0.96 +/- 0.14 mmol/l, magnesium diasporal 0.843 +/- 0.14 to 0.891 +/- 0.14 mmol/l and chemiron 0.848 +/- to 0.866 +/- 0.16 mmol/l after five days. The ferrous gluconate and folic acid treated group showed no significant changes. This study shows that a chemiron supplement leads to increased magnesium plasma levels whereas ferrous gluconate and folic acid do not. These results suggest that the low level of magnesium is a normal physiological adjustment of pregnancy and that iron supplementation does not influence this unless magnesium salt is given. PMID- 9743889 TI - A medical method of early pregnancy termination using tamoxifen and misoprostol. AB - A study was undertaken to determine whether ingestion of the selective estrogen receptor modulator tamoxifen followed by vaginal administration of the prostaglandin misoprostol would be an effective medical method of elective termination of early pregnancy. A clinical trial was conducted with a study group of 100 healthy women with pregnancies of 56 days gestational age or less who desired elective pregnancy termination. Each subject ingested 20 mg of tamoxifen once daily for 4 days followed 4 days later by intravaginal placement of four 200 micrograms tablets of misoprostol. If abortion did not occur within the next 24 h a second dose of 800 micrograms of misoprostol was given. The main outcome measures were incidence of complete abortion, hemoglobin levels, duration of vaginal bleeding, and incidence of side effects. Complete abortion occurred in 92 (92%, 95% CI 86.7, 97.3%) of 100 subjects. Of these 92 women, four aborted after ingesting tamoxifen without use of misoprostol, 84 within 24 h after receiving a single dose of misoprostol, one 21 days following a single dose of misoprostol, and three after a second dose of misoprostol was administered. There were six (6.0%) complete treatment failures and two (2%) incomplete abortions that required a dilatation and curettage. The mean duration of uterine bleeding was 8.1 days (range 1-34 days) and there was a median decrease in hemoglobin level of 0.50 g/dL (+2.2 to -4.7 g/dL). Vomiting occurred in 28% of subjects and diarrhea in 8%. These initial data suggest that ingestion of tamoxifen followed by intravaginal misoprostol may be an effective, easily administered, and inexpensive method to electively induce complete abortion in pregnancies of 56 days gestational age or less. Additional studies are necessary to determine whether the addition of tamoxifen increases the success rate compared with that obtained with the use of vaginally administered misoprostol by itself. PMID- 9743890 TI - Introduction of cyclofem once-a-month injectable contraceptive in Mexico. AB - A large introductory study of Cyclofem, a once-a-month injectable contraceptive, was conducted in three Mexican provinces. A total of 3457 healthy women participated: 640 women from rural areas (community-based component) and 2817 women from urban and suburban areas (health center-based component). A total of 20,316 women-months of treatment experience were accumulated during a one year period. Cyclofem proved its use-effectiveness (pregnancy rate of 0.03%) and its safety under routine service conditions of family planning facilities in Mexico. The overall life table continuation rate at 1 year was 26.1%. Higher continuation rates were observed in the community-based component (36.6%) as compared to the health center component (23.7%). The most common reason for method discontinuation was change of address. Only 15% of the discontinuations were attributable to the injectable contraceptive method, with the overall 1 year discontinuation rate for bleeding problems (including amenorrhea) was < 11%. These observations underscore the importance of appropriate counseling and follow up measures, providing convenient access to repeat injections, and other service delivery issues related to continuation of Cyclofem. The results of this trial have once again demonstrated that Cyclofem is a highly effective method with an acceptable side effect profile. In addition, the study provided the elements for its approval by local health authorities and its inclusion into the Ministry of Health Family Planning Program. PMID- 9743891 TI - A scenario study of oral contraceptive use in Japan. Toward fewer unintended pregnancies. AB - A scenario study was conducted to assess the extent to which the unintended pregnancy rate in Japan, where oral contraceptives (OC) have not been legalized for family planning purposes and couples rely mainly on condoms, might change if more women were to use OC. Because current rates of unintended pregnancy and abortion in Japan are not known, data provided by the 1994 Japanese National Survey on Family Planning were used to construct scenarios for national contraceptive use. Annual failure rates of contraceptive methods and nonuse were applied to the contraceptive use scenarios, to obtain estimates of the annual number of contraceptive failure-related pregnancies. Subsequently, contraceptive practice situations assuming higher OC use rates were defined, and the associated change in the number of contraceptive failure-related pregnancies was estimated for each situation. It emerged that OC use rates of 15% decreased the expected number of unintended pregnancies by 13%-17%, whereas use rates of 25% resulted in decreases of 22%-29% and use rates of 50% in decreases of 45%-58%. The findings were reasonably robust to variation in the assumptions that were made. In conclusion, each theoretical percentage increase in the OC use rate in Japan was found to lead to a roughly equivalent percentage decrease in the number of unintended pregnancies. PMID- 9743892 TI - Vaginal bleeding patterns in users of Perlutal, a once-a-month injectable contraceptive consisting of 10 mg estradiol enanthate combined with 150 mg dihydroxyprogesterone acetophenide. A trial of 5462 woman-months. AB - Perlutal (other names: Topasel, Perlutan) is a once-a-month injectable contraceptive that contains 10 mg estradiol enanthate and 150 mg dihydroxyprogesterone acetophenide. A prospective trial was conducted in 216 women in Medellin, Colombia, over five years (5,462 woman-months) to establish the rates of the different vaginal bleeding patterns during the use of Perlutal, and to assess their relation with discontinuation of the Perlutal regimen. It was found that with the use of Perlutal, the duration of menstrual bleeding decreased from 3.9 to 2.7 days (p < 0.01), and that the incidence of dysmenorrhea decreased from 31% to 1.6% (p < 0.01). At one year of follow-up, the incidence of altered bleeding patterns was 5.1%. The discontinuation rate due to an altered bleeding pattern was 3.9%. It is concluded that the low incidence of altered bleeding patterns observed with the use of Perlutal leads to a low discontinuation rate among users. PMID- 9743893 TI - Prevalence and determinants of current contraceptive method use in a palm oil company in Cameroon. AB - The principal reasons given by African women for not using contraception include their lack of economic power and control over their choice of partner. An epidemiologic descriptive survey of a cross-section of the female personnel of a Cameroonian palm oil company (SOCAPALM) was carried out in August 1995, to evaluate the various determinants and level of use of various family planning methods in a well defined population of women in employment. An exhaustive list of all the households in the five villages of SOCAPALM was compiled and all women between 15 and 49 years of age who had lived on the palm oil plantation for at least a year were interviewed. The adjusted odds ratios showed that use of modern contraceptive methods was significantly associated with the woman having received secondary education, having more than three children, being the head of the household and, in cases where there was a man regularly present in the household, his approval of family planning. Recently receiving information (during the last month) about family planning was not identified by multivariate analysis as a significant factor affecting the decision to use modern or traditional contraception. The same factors were found to be associated with the use of traditional methods of contraception, but having had an illegal abortion was also associated with the use of such methods. Thus, the level of knowledge about family planning and the prevalence of contraceptive use was significantly higher for women living in industrial environments (such as SOCAPALM), than in the overall population of women in Cameroon. The economic power of the woman, the presence of a strong social reproductive health network, and the positive attitude of men and community leaders were the most important factors affecting the family planning decision of the women. PMID- 9743894 TI - Dose-response effects of gramicidin-D, EDTA, and nonoxynol-9 on sperm motion parameters and acrosome status. AB - Previous reports showed that gramicidin-D (G-D), a polypeptide with antiviral and antimicrobial properties, nonoxynol-9 (N9), a common spermicidal detergent, and EDTA, a Ca-Mg chelating agent, inhibited sperm motility and cervical mucus penetration. The purpose of this study was to determine the dose-response effects of G-D, N9, EDTA and G-D + EDTA on sperm motion parameters and acrosome status. Semen specimens from known fertile donors were subjected to computer-assisted semen analysis of motility, path velocity, progressive velocity, and hyperactivation prior to and after incubation with varying concentrations of gramicidin-D, EDTA and nonoxynol-9. Each specimen was also prepared for acrosome status using rhodamine isothiocyanate conjugated pisum sativum agglutinin (RITC PSA). There was a significant decrease in motility by G-D, EDTA, G-D + EDTA, and N9 at all doses as compared to the fresh specimen. N9 completely immobilized all sperm at each dose. Progressive velocity and path velocity also decreased in a dose-response manner. Sperm hyperactive motility also significantly decreased in all groups. The majority of sperm remained acrosome intact following exposure to all doses tested, whereas N9 resulted in complete breakdown/release of the acrosomal contents. This study confirms previous reports that G-D, EDTA, and N9 significantly impair sperm motility and motion parameters. The effective 100% inhibitory concentration was seen only with N9, whereas G-D, EDTA, and G-D + EDTA resulted in incomplete impairment of sperm motion parameters. At the concentrations used, N9 demonstrated potent spermostatic activity. Gramicidin-D and EDTA should be further studied for their potential contraceptive spermostatic activity. PMID- 9743895 TI - Safety, tolerability, and pharmacokinetics of a novel, selective antiprogestagen (Org 31710) in healthy male volunteers. AB - The safety and tolerability as well as pharmacokinetics of a new selective antiprogestagen, Org 31710, were studied after oral administration of single doses of 10, 25, 50, or 75 mg to 24 healthy male volunteers. Per dose-group, five subjects received active and one subject received placebo treatment. In subjects receiving 75 mg, the effects of Org 31710 on serum levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), and testosterone were also studied. No adverse or seriously adverse events were observed. All doses of Org 31710 were well tolerated. Characterization of the Org 31710 plasma pharmacokinetics revealed a statistically significant deviation from linearity: the dose normalized Cmax (nCmax) and dose normalized area under the curve (nAUC) values were significantly lower for the higher dosages (p < 0.05). Furthermore, tmax tended to decrease (from 1.6 to 0.9 h), whereas the elimination half-life (t1/2) tended to increase (from on average 45 to 57 h) with increasing dose. Org 31710 did not have any effect on serum levels of FSH, LH, and testosterone. In conclusion, Org 31710 appears to be a safe and well-tolerated compound in the dosage range studied. PMID- 9743896 TI - Screening for antiproliferative actions of mifepristone. Differential endometrial responses of primates versus rats. AB - This laboratory has previously shown the capability of the antiprogestin, mifepristone, to noncompetitively inhibit estrogen-induced endometrial proliferation in nonhuman primates. In the following study, use of the rat uterine weight bioassay was compared against a primate (Macaca fascicularis) uterine bioassay to identify the noncompetitive/antiproliferative effects of mifepristone. These uterine bioassays were contrasted for reasons of identifying a comparative laboratory rodent model that could substitute for the need to use primate models in the screening of potential antiprogestins, thereby saving time, cost, and primate resources. Results of the primate experiment showed that mifepristone decreased endometrial proliferation in a dose-dependent manner; importantly, this decrease occurred in the presence of sustained physiologic serum 17 beta-estradiol (E2) levels. However, in the rat model, results showed that mifepristone altered uterine wet weight and blotted weight values only in those animals receiving pharmacological doses of E2 (p < 0.05). Based on the results summarized herein, use of this rat uterine weight bioassay as a substitute for primate models is not recommended for screening and identification of "interesting" antiprogestins. Apparently, the endometrial noncompetitive antiestrogenic/antiproliferative effects of mifepristone, observed repeatedly in these laboratory primates, do not operate in the rat uterine tissue. PMID- 9743897 TI - Contraceptive testing of vaginal agents in rabbits. AB - Development of new vaginal products, such as microbiocides and contraceptives, requires in vivo testing of their effect on fertility. Rabbits, unlike smaller laboratory animals such as rats and mice, which inseminate in the uterus, inseminate vaginally and thus are valuable as animal models for testing vaginal agents for contraceptive effects. Rabbits are inexpensive and easy to handle compared to nonhuman primates, and have frequently been used for testing the effects of vaginal agents on fertility. We review the pertinent literature and report findings that provide guidance for effectively using and improving the rabbit contraceptive model in testing new vaginal products. PMID- 9743898 TI - Uterine luminal epithelial alkaline phosphatase activity and pinopod development in relation to endometrial sensitivity in the rat. AB - The period of maximal endometrial sensitivity in the rat was characterized by high alkaline phosphatase activity in uterine luminal and glandular epithelium and endometrial stroma. The activity in endometrial stroma increased following decidualization. Pinopod development on the endometrial surface was first observed during the presensitivity period. Their number increased, apparently more so on the antimesometrial rather than the mesometrial segment of the uterus, on the day of maximal sensitivity. Inhibition in endometrial sensitivity by single anti-implantation (1.25 mg/kg, po) dose of centchroman on day 1 post coitum (p.c.), although it did not affect alkaline phosphatase activity on days 2 and 3 p.c., caused complete inhibition in its activity in uterine luminal and glandular epithelium and pinopod development on days 4 and 5-coinciding, respectively, with time of entry of preimplantation embryos into the uterus and period of maximal endometrial sensitivity in this species. Significant decrease in enzyme activity was also evident in the entire endometrial stroma and myometrium, except blood capillaries, on these days. In comparison, prevention of entry of native embryos into the uterus by placing a ligature at the utero-tubal junction had no effect on pinopod development, but caused marked decrease in enzyme activity in luminal and glandular epithelium only during the immediate postimplantation period. The uterine lumen on the day of maximal sensitivity in centchroman-treated rats appeared highly distended and was lined with tall columnar epithelium, in comparison to low cuboidal epithelium in controls. The findings demonstrate: (a) a correlation between uterine luminal epithelial alkaline phosphatase activity and endometrial sensitivity; (b) complete inhibition in enzyme activity in luminal and glandular epithelium following centchroman treatment might be related to altered permeability characteristics of epithelial cells, which together with the absence of pinopods and highly distended uterine lumen on the day of maximal sensitivity, suggest inhibition of endocytosis/pinocytosis of luminal fluid, luminal closure, apposition of blastocyst trophoblast to luminal epithelium, and secretory activity of glandular epithelium; (c) pinopod development on the endometrial surface was independent of presence of viable blastocysts in utero; and (d) complete absence of pinopods suggests lack of endometrial sensitivity, but their presence might not necessarily indicate a sensitized endometrium in the rat. PMID- 9743899 TI - ECRI to clinical engineering departments: "heads up! ... FDA's potential regulation of servicers, remarketers, and refurbishers could affect you". PMID- 9743900 TI - Medical device problem reporting for the betterment of healthcare. AB - Given that there are nearly 5,000 individual classes of medical devices, tens of thousands of medical device suppliers, and millions of healthcare providers around the world, device-related problems are bound to happen. But effective problem reporting can help reduce or eliminate many of these problems--not only within an institution, but also potentially around the world. In this article, we trace the problem reporting process from its beginnings in the hospital to its global impact in making critical information available throughout the healthcare community. PMID- 9743901 TI - Digital film processing: a comparison of wet and dry processing methods. AB - Traditional film-based imaging has always relied on film processing that uses liquid chemicals--a method sometimes referred to as wet film processing. As digital imaging has become available for modalities ranging from radiography to magnetic resonance imaging, however, a digital-based processing method called dry film processing has been devised. Dry film processing transfers digital information to film using heat or a combination of heat and light, and the images are processed by the films themselves. In this article, we present an overview of each film processing method, along with a comparison between the methods as they are used by laser imagers and other imager types. PMID- 9743902 TI - The Laser Institute of America's laser hazard evaluator. AB - The Laser Institute of America recently developed a software program, the Laser Hazard Evaluator, to help analyze the risks involved in using most types of lasers. In this article, we review the uses and characteristics of the Laser Hazard Evaluator and present ECRI's judgment of its merits. PMID- 9743903 TI - Sparking from and ignition of damaged electrosurgical electrode cables. PMID- 9743904 TI - [Significant role of penem antibiotics: focused on faropenem (discussion)]. PMID- 9743905 TI - [The drug sensitivity of enterohemorragic Escherichia coli and antibiotics treatment for hemorrhagic enterocolitis--from an outbreak of enterocolitis in Sakai city]. AB - In July, 1996, a massive outbreak of hemorrhagic enterocolitis involving more than 5,000 people was caused by enterohemorragic Escherichia coli (EHEC) O157:H7 occurred mainly among elementary school children of Sakai City, Japan. The antibacterial activities in vitro against EHEC from stool specimens were determined. Norfloxacin showed the highest antibacterial activity, and fosfomycin, kanamycin, ampicillin, cefaclor were considered as effective drugs. But doxycycline showed lower antibacterial activities compared to other examined drugs, and it appears necessary to take antibiotic resistance of Escherichia coli into consideration when a treatment regimen is determined. As a result of the oral administration of fosfomycin to 95 patients of hemorrhagic enterocolitis and carrier, no patients developed complications with hemolytic uremic syndrome (HUS). However, a study of 17 patients with HUS demonstrated the fact that most of them were subjected to intravenous administration of fosfomycin. It may be needed to consider oral administration route of effective antibiotics in the treatment of enterocolitis in order to maintain high concentrations of a drug in the intestine. PMID- 9743906 TI - [Pharmacokinetic and clinical studies with cefluprenam in the pediatric field. Pediatric Study Group of Gefluprenam]. AB - Evaluation of efficacy and safety of cefluprenam (code number: E1077, abbreviation: CFLP), a newly developed injectable cephem antibiotics was conducted on adult patients with various infections, and followed by the study group organized from 39 institutions in pediatric field, as the drug showed no toxicity problems in suckling animals. Informed consents from legal representatives were obtained prior to the study. 1. Clinical efficacy. Two hundred eighty one cases were included for analysis of clinical efficacy after 40 cases of exclusion or drop-out were subtracted from a total of 321 cases. However, the cumulative number of cases evaluable for analysis was considered to be 289, because 8 cases that had 2 different diseases at the same time were counted in each category of disease. In the cases in which causative organisms were identified (group A), 148 of 154 cases were rated as good or excellent, with an efficacy rate of 96.1%. As for clinical efficacies by disease, efficacy rates were 6/6 for purulent meningitis, 4/5 for sepsis, 95.7% (62/65) for pneumonia, 100.0% (29/29) for urinary tract infections, and 94.1% (16/17) for skin and soft tissue infections. The rate of excellent responses among excellent and good responses was 73.6% (109/148), showing a higher value than any of recent injectable beta-lactams. On 32 cases with S. pneumoniae infection, the efficacy rate of CFLP was 100.0%. In the cases where causative organisms were not identified (group B), 128 of 135 cases were rated as good or excellent, with an efficacy rate of 94.8%. In the all cases including both the group A and the group B, the efficacy rate was 95.2% (276/289) and the rate of excellent responses among excellent and good response was 70.7% (195/276). Against severe infections, CFLP exhibited excellent clinical efficacy, showing an efficacy rate of 8/8 for meningitis, 3/5 for sepsis and 100.0% (22/22) for severe pneumonia. As for bacteriological responses, eradication rates were 95.2% (177/186) in total. Against Gram-positive cocci, the eradication rate was 92.7% (76/82), with eradication rates of 94.3% (33/35) for Staphylococcus aureus, and 93.3% (28/30) for Streptococcus pneumoniae. Against Gram-negative rods, the eradication rate was 97.1% (101/104), and eradication rates were 100.0% (22/22) for Escherichia coli, 97.5% (39/40) for Haemophilus influenzae and 100.0% (19/19) for Molaxella catarrhalis. In cases in which more than 3 days of treatment with previous chemotherapy resulted in no response, the efficacy rate of CFLP was 94.2% (98/104), rated excellent in 68 cases and good in 30 cases. In these cases, the eradication rate was 98.1% (52/53). 2. Pharmacokinetics. CFLP was intravenously administerrd to 12 subjects at doses of 20 to 40 mg (potency)/kg. In 9 subjects aged more than 12 months, maximum serum levels (Cmax), T 1/2 beta and AUC of CFLP were 155.3 +/- 9.8 micrograms/ml, 1.43 +/- 0.18 hours and 111.7 +/- 15.0 micrograms.hr/ml, respectively, when a dose of 20 mg (potency)/kg was used. In 2 subjects aged not more than 12 months, the mean Cmax, T 1/2 beta and AUC were 153 micrograms/ml, 1.6 hour and 81 micrograms.hr/ml, respectively, at a dose of 20 mg(potency)/kg. The mean Cmax, T 1/2 beta and AUC were 332 micrograms/ml, 0.93 hours and 157.3 micrograms.hr/ml, respectively, in 1 subject at a dose of 40 mg (potency)/kg. In 10 subjects dosed 20 mg (potency)/kg, urinary levels were 2413 +/- 512, 1471 +/- 524, and 470 +/- 115 micrograms/ml in 0-2, 2-4, and 4-6 hours after dosing, respectively, showing a cumulative urinary excretion rate of 61.4 +/- 6.3%. In 1 subject dosed 40 mg (potency)/kg, urinary levels were 5700 and 4770 micrograms/ml in 0-2 p3d 2-4 hours after dosing, respectively, showing a cumulative urinary excretion rate of 42.1%. Cerebrospinal fluid concentrations of CFLP, on 10 subjects with purulent meningitis dosed 40-103 mg (potency)/kg were 3.2-32.9 micrograms/ml at 0.5-2 hours after administration within 4 days after the onset of PMID- 9743907 TI - [The clinical efficacy of imipenem/cilastatin sodium in orthopedic infections and drug levels in the bone tissue]. AB - We evaluated the clinical efficacy of imipenem/cilastatin sodium (IPM/CS--a carbapenem antibiotic) against orthopedic infections, and the drug levels of the bone tissues were determined. The clinical efficacies for 6 patients in the infection group were good in 3 cases, and fair in the other 3; giving an efficacy rate of 50%. Bacteriologically, 8 strains were isolated from patients with the infection and an eradication rate of 87.5% was obtained upon the treatment. In 39 patients that were given the drug prophylactically, no postoperative infections occurred. Mean IPM levels in the bone and the bone marrow at 1 hour after administration in 5 patients of the prophylactic group were 17.3 micrograms/ml and 5.9 micrograms/g, respectively. The ratio of concentrations the bone to those in the bone marrow was 34.6%. The results of this study suggest that IMP/CS reaches to the bone tissue providing sufficient concentrations and that the drug is efficacious for the prophylaxis and the treatment of orthopedic infections. PMID- 9743908 TI - [Efficacy and safety of intramuscular imipenem/cilastatin (IPM/CS) for complicated urinary tract infections]. AB - An intramuscular preparation of imipenem/cilastatin (IPM/CS, 500 mg/500 mg) was administered to 59 patients with complicated urinary tract infections (UTI; cystitis and pyelonephritis) to evaluate its efficacy and safety. The obtained results are summarized as follows: In patients with cystitis, evaluations based on daily frequencies of administration were also performed. 1) According to the treating doctors, the drug showed an overall efficacy rate of 80% (45/56 patients). The efficacy rate was 89% in patients with cystitis treated by a u.i.d. regimen. Among patients treated by a b.i.d. regimen, the efficacy rate was 67% for cystitis cases and 84% for pyelonephritis cases. 2) When clinical efficacy was assessed according to the criteria for UTI drug efficacy evaluation, the drug was 'markedly effective' in 14 patients, 'effective' in 23, and ineffective in 11 patients, for an efficacy rate of 77% (37/48 patients). 3) The microbiological eradication rate was 88% (59/67 strains). The rate was 95% (20/21 strains) for Gram-positive bacteria and 85% (39/46 strains) for Gram-negative bacteria. The efficacy for Enterobacter faecalis and Pseudomonas aeruginosa was 100% and 73%, respectively. 4) As side effects, pain at the injection site was reported by one patient and abnormal laboratory test values were observed in 2 patients. All of these reactions were mild and resolved shortly after the completion of treatment. Based on these findings, it is concluded that this intramuscular preparation of IPM/CS is effective for treating complicated urinary tract infections. PMID- 9743909 TI - [Antimicrobial activities of roxithromycin against recently obtained clinical isolates]. AB - The purpose of our investigation was to monitor current trends in the susceptibility patterns of clinical bacterial isolates to roxithromycin (RXM). We measured the MICs of macrolide antibiotics, such as RXM, erythromycin (EM), clarithromycin (CAM), rokitamycin (RKM) and midecamycin (MDM), and other classes of antibacterial compounds against various clinical isolates at seven institutions between October and December in 1994 and 1995. RXM had excellent antibacterial activities for S. pyogenes, S. agalactiae, M. (B.) catarrhalis and methicillin sensitive S. aureus. Against methicillin sensitive S. epidermidis, RXM activity was fairly good but about 20% of the strains had MIC > or = 128 micrograms/ml. The activity against S. pneumoniae was not so potent and similar to activities of EM, CAM, MDM, and clindamycin. The vast majority of methicillin resistant S. aureus and S. epidermidis were also resistant to macrolide antibiotics and other classes of compounds tested. In conclusion, RXM is an unique macrolide antibiotic by retaining potent activity against S. pyogenes, S. agalactiae, S. aureus except MRSA, M. (B.) catarrhalis and M. pneumoniae. PMID- 9743911 TI - Influence of additives on the release profile of nifedipine from poly(DL-lactide co-glycolide) microspheres. AB - Nifedipine-containing poly(DL-lactide-co-glycolide) (PLGA) microspheres of various sizes and drug contents were prepared by the solvent evaporation method. The in vitro release profiles of nifedipine from PLGA microspheres and the degradation pattern of the polymer were evaluated. Four additives were incorporated in the microspheres: two non-fatty plasticizers: diethylphthalate and triacetin, and two fatty substances: isopropyl myristate and Myvacet. Diethylphthalate and Myvacet increased the nifedipine release rate while isopropyl myristate and triacetin had no influence on it. Triacetin seems to be very poorly incorporated into the microspheres. These additives did not modify the degradation rate of the polymer. Differential scanning calorimetry detected a decrease of the glass transition temperature of diethylphthalate-containing microspheres, a small variation with Myvacet, and very little change when triacetin or isopropyl myristate were incorporated. This variation of the glass transition temperature (Tg) tends to imply that nifedipine is released by a diffusion process through the polymer matrix which is enhanced when additives decrease the Tg. Scanning electron microscopy allowed the vizualization of the highly porous structure of microspheres containing the oily substances, and the unchanged smooth surface of diethylphthalate-containing microspheres. PMID- 9743912 TI - The physical and chemical stability of suspensions of sustained-release diclofenac microspheres. AB - The major challenge in liquid sustained-release oral suspensions is to minimize drug diffusion into the suspending medium and to retain the original properties of the microparticles during storage. Diclofenac wax microspheres prepared by the hydrophobic congealable disperse phase method were formulated as a sustained release suspension and stored at three different temperatures (25, 37 and 45 degrees C) for 3 months, to evaluate the physical and chemical stability of the suspended microspheres. Suspensions of microspheres stored at ambient temperatures were both physically and chemically stable, but at higher temperatures, up to 45 degrees C, there was a decrease in drug release due to scaling and melting on the microsphere surface as observed by scanning electron microscopy. However, on prolonged storage, up to 90 days, especially at 45 degrees C, temperature became a dominant factor causing an increase in drug release. The suspension of diclofenac microspheres was chemically stable for 3 months, while the plain drug suspension exhibited slight degradation. PMID- 9743913 TI - beta-Glucuronidase activity following complex coacervation and spray drying microencapsulation. AB - The objective was to develop a microencapsulation process suitable for the controlled release of an active protein drug. beta-glucuronidase was selected as a model protein and a combination of complex coacervation (gelatin/sodium alginate, gelatin/acacia and albumin/acacia) and spray drying was investigated. Coacervates were either spray dried or glutaraldehyde crosslinked to form microcapsules. Polyvinylpyrrolidone (PVP) and polyethylene glycol were investigated as potential coacervate enhancers and stabilizers. beta glucuronidase/polymer mixtures were spray dried to determine any polymer protective effects on protein activity. A BUCHI 190 Spray Drier was used, beta glucuronidase activity was determined using a Sigma Kit and microcapsule particle size was measured by Accusizer analysis (light blockage). All non-crosslinked coacervates investigated, with the exceptions of albumin/acacia and albumin/acacia/beta-glucuronidase/PVP, were unsuitable for spray drying as they rapidly phase separated and blocked the spray drier nozzle. beta-glucuronidase activity in the albumin/acacia coacervates approximated to 99% prior to and 80% following spray drying. This can be compared to activities of approximately 30% and 68% when spray dried alone and with albumin, respectively, and of 18% in albumin/acacia microcapsules crosslinked with glutaraldehyde. Microcapsule particle size was affected by coacervation pH, additives and spray drying. In vitro beta-glucuronidase release was biphasic, with an initial burst release followed by a zero order release phase and continued over the 12 day study period. In conclusion, the spray drying albumin/acacia/PVP method described is useful for the preparation and collection of controlled release microcapsules with minimal loss of beta-glucuronidase activity. PMID- 9743914 TI - Methotrexate loaded chitosan and chitin microspheres--in vitro characterization and pharmacokinetics in mice bearing Ehrlich ascites carcinoma. AB - Methotrexate (MTX) loaded chitosan and chitin microspheres were prepared. The physicochemical characteristics were affected by various parameters, e.g. stirring speed, concentration of chitosan and chitin. The t1/2 for in vitro release varied depending on the concentration of chitosan and chitin and the amount of glutaraldehyde used. The antitumour activity in Ehrlich ascites tumour bearing mice for MTX loaded chitosan microspheres was better when compared with plane MTX on oral administration. The plasma drug levels were sustained. PMID- 9743915 TI - Adsorption of chitin derivatives onto liposomes: optimization of adsorption conditions. AB - A 2(4) factorial design is used to optimize the adsorption conditions of the hydrophilic anionic polyelectrolytes. Carboxymethylchitin (CMC) and Carboxymethyl/Glycolchitin (CO) onto liposomes at physiological ionic strength (I) and pH using phosphate buffered saline (PBS, I = 154 mM, pH = 7.4). Positive ([+]) or high surface affinity liposomes (DSPC:CHOL:DMTAP, 5:4:1), and Neutral ([N]) or low surface affinity liposomes (DSPC:CHOL, 1:1) were used as adsorption surfaces. Results of the calculations of the main effects indicate that polymer molecular weight (mwt), Surface Affinity (S), Number of Adsorption Shots (Sh), Temperature (T), and the combinations mwt x S, mwt x Sh are the most important process parameters. Results of a study conducted at T = 37 degrees C show that no loss occurs from the positive surface at the highest particle concentration, Np = 4.043 x 10(11). Finally, the extent of polymer-induced particle aggregation is decreased when the diameter of the uncoated liposomes is doubled from 0.22 to 0.45 micron. These results are as expected, given the stiffness and the dimensions of the macromolecules. PMID- 9743916 TI - Slow release polymer-drug systems obtained by moisture promoted polyreactions. 1. Codeine resinate encapsulated in poly(alkyl alpha-cyanoacrylates). AB - Samples of codeine resinate consisting of a carboxylic cation exchanger (as a polymeric carrier) and codeine (as a drug) were coated with poly(alkyl alpha cyanoacrylates) by suspending and stirring wet resinate beads in a toluene solution of the monomer. Methyl, ethyl and n-butyl alpha-cyanoacrylates were used as monomers. Each coated material released codeine more slowly than the non coated. The data obtained confirmed that water promoted the polymerization of alkyl alpha-cyanoacrylates, which proceeded at the surface of the wet resinate beads. The rate of codeine release depended on the type of monomer used for coating, the monomer/resinate feed ratio and the plymerization time. PMID- 9743917 TI - Preparation and tabletting of dipyridamole alginate-Eudragit microspheres. AB - Dipyridamole alginate-Eudragit microspheres were prepared by the emulsification technique of Monshipouri and Price. After modification of the formulation and process variables of this method it was possible to obtain spherical and discrete particles. Microspheres were free flowing and the release of drug was slow compared to granules and powder. To further control the release rate of the drug, tablets of microspheres were prepared, adding 20% HPMC 90 SH 4000 and 90 SH 100,000 and compressing at 98 mPa or 147 mPa. Increasing the amount of additive and the compression pressure decreased the release rate of drug. PMID- 9743919 TI - Microencapsulation of rhizobacteria by spray-drying: formulation and survival studies. AB - This research deals with the microencapsulation of the bacteria Pseudomonas fluorescens-putida, using the spray-drying technique. These bacteria act as a plant growth stimulator and the microparticles are subsequently included in the seed coating or pelleting material. The objective was to maintain living encapsulated bacteria for a minimum of 5-6 months. Three polymers were tested: a methacrylic copolymer from the Eudragit range, an ethylcellulose and a modified starch. A silica additive to the spraying food was also examined. The granulometric distribution and the morphology of the microparticles were studied and the residual moisture was measured after each bacteria survival control test. The best results were obtained with the methacrylic copolymer Eudragit, particularly for microparticles collected in the cone of the spray-drying chamber. The mean diameter of the microspheres was 44.6 microns, 85% of these particles having a size between 6.2 and 84.4 microns. The bacterial survival time of a particular strain incorporated in these microparticles, strain M3.1, was one year. A relationship was found between the bacterial survival and the microspheres' residual moisture, the best survival of the stored bacteria being observed when the residual moisture was around 25%. PMID- 9743918 TI - Preparation and characterization of beta-cyclodextrin and poly(acrylic acid) microspheres. AB - A method for the preparation of poly(acrylic acid) (PAA) microspheres crosslinked with beta-cyclodextrin (beta-CD) is described. The method is based on a water-in oil (w/o) emulsion solvent evaporation technique which facilitates a condensation reaction between PAA and beta-CD. Aqueous solutions of PAA and beta-CD were used as the dispersed phase and food grade olive oil was used as the continuous phase. The effect of homogenization speed (used in the preparation of the emulsion), phase volume ratio and cyclodextrin-polymer load on the particle size of the microspheres produced was investigated in a replicated factorial design. Microspheres were sized by light microscopy. The particle size of the microspheres was influenced by all three variables with two significant first order interactions between the variables being observed (homogenization speed with phase volume ratio and homogenization speed with load). A second order interaction between the three principal factors was also observed. Particle size ranged from 16 to 150 microns, depending on the production variables employed. The yield for the technique was 69.5 +/- 9.5%. Using selected conditions, microspheres of 15-25 microns size were prepared from a range of PAA with different weight average molecular weights (Mw). These particles were then characterized for beta-CD, free carboxylic acid group content and residual oleic acid. PMID- 9743920 TI - Effects of preparation conditions on the monodispersity of albumin microspheres. AB - Monodisperse albumin microspheres were successfully prepared by both chemical or thermal hardening methods via membrane emulsification using microporous glass membranes with uniform pore sizes. The monodispersity of the microspheres was found to depend strongly on parameters such as albumin concentration, emulsifier concentration, and volume ratio of the internal aqueous phase (albumin solution) to the dispersion medium (organic solvent). The optimum conditions for obtaining monodisperse albumin microspheres are described. PMID- 9743921 TI - Influence of dextran molecular weight on capture in and release from decylamine carboxymethylcellulose capsules. AB - A series of dextran molecular weight markers were encapsulated in decylamine carboxymethylcellulose microcapsules to serve as probes of capsule retentivity. The capsules were prepared by allowing microdrops of aqueous sodium carboxymethylcellulose to fall into aqueous decylamine acetate solution. Salt exchange reaction at the droplet pseudointerface resulted in self-assembling films which essentially instantaneously enclosed the droplets. Concentrations of anionic polymer were varied in the range from 1-3%. Chromophore-bearing dextrans were incorporated into these capsules by blending the dextrans with the carboxymethylcellulose prior to the encapsulation step. Four dextrans of differing (light scattering) molecular weights were used: 2 x 10(6), 6 x 10(5), 7 x 10(4), and 1.9 x 10(4) amu. The mass balance of dextran retained in the capsules, released on washing the capsules or which escaped encapsulation was determined spectrophotometrically. To measure total dextran in a population of washed capsules, the capsules were lysed in a 0.3 M solution of sodium chloride. To monitor dextran release, washed capsules were suspended in water and dextran concentration in the supernatant was measured. Encapsulation efficiency exceeded 80% for high molecular weight dextran but was lower with the smaller dextrans. PMID- 9743922 TI - Abstinence--more than a slogan. PMID- 9743923 TI - Parents' reports of "tricks of the trade" for managing childhood chronic illness. AB - PURPOSE: To examine how parents respond to and manage the challenges of childhood chronic illness. DESIGN: Qualitative, comparative, secondary analysis. PARTICIPANTS: Fifty-eight families (55 mothers, 44 fathers) with a school-age child (6-15 years old) with a chronic illness. RESULTS: Three approaches to illness management were identified: strict adherence, flexible adherence, and selective adherence. These approaches varied in the extent to which parents developed and relied on target management behaviors that concurred with or deviated from the treatment plan prescribed by healthcare providers. CONCLUSIONS: The three management approaches contribute to understanding the processes associated with differing interactions between healthcare professionals and parents when a child has a chronic illness. PMID- 9743924 TI - Stress and coping behaviors of substance-abusing mothers. AB - PURPOSE: To examine parenting stress and coping behaviors in substance-abusing and non-substance-abusing mothers. DESIGN: A comparative descriptive design. SETTING: Pediatric primary care clinic. PARTICIPANTS: Low-income, predominantly African-American mothers (N = 60) of young children recruited from a pediatric primary care clinic. Thirty mothers were known substance abusers and 30 had no known history of substance abuse. MAIN OUTCOME MEASURES: Parenting Stress Index/Short Form (PSI/SF) and Child Protective Service (CPS) validation of abuse or neglect. RESULTS: Substance-abusing mothers scored significantly higher than comparison mothers on total stress and the three subscales of the PSI/SF: parent child dysfunctional interaction, difficult child, and parental distress. Forty seven percent of substance-abusing mothers scored in the clinical range on total stress compared with only 3.3% of non-substance-abusing mothers. Proportionately more substance-abusing mothers than comparisons demonstrated maladaptive parenting behaviours as evidenced by CPS-confirmed abuse or neglect of their youngest child. CONCLUSIONS: Substance-abusing mothers of young children are at increased risk for increased levels of stress and maladaptive coping behaviors. Substance-abusing mothers need support and monitoring in the parenting role and referrals to substance-abuse and parenting programs. PMID- 9743926 TI - The effectiveness of a standardized educational program for children using patient-controlled analgesia. AB - PURPOSE: To examine the effectiveness of two types of preoperative education (routine education and a standardized educational program) for children undergoing spinal fusion. DESIGN: Two group, phase-lag design. SETTING: Tertiary pediatric hospital. PARTICIPANTS: Children ages 8-18 years (N = 93). MAIN OUTCOME MEASURES: Adolescent Pediatric Pain Tool, Child Pain Scale, Post-PCA Satisfaction Interview, and PCA infusion pump data. RESULTS: No statistically significant differences between the groups on any of the main outcome variables. Children and parents reported, however, that the SEP provided them with invaluable information regarding the use of PCA and alleviated their concerns about getting "hooked on drugs," overdosing, side effects, and being able to get pain relief when needed. CONCLUSION: Children having spine fusion surgery experienced severe postoperative pain that was not ameliorated by optimizing use of PCA through standardized education. Further testing of the SEP with other populations is needed in order to more fully realize its potential for influencing pain outcomes. PMID- 9743925 TI - A randomized trial of heparin and saline for maintaining intravenous locks in neonates. AB - PURPOSE: To determine the effects of saline, heparin 2 units (U) per ml saline, and heparin 10 U/ml saline flush solutions on the duration of intravenous (i.v.) locks and the incidence of i.v. infiltration in neonates. DESIGN: Randomized double-blind experiment. SETTING: Tertiary-care nursery. PARTICIPANTS: Neonates (N = 90) hospitalized at birth in the intensive, intermediate care, or newborn units. MAIN OUTCOME MEASURES: Total hours from the time the i.v. was inserted to the time the i.v. was removed; hours from the time the i.v. was first flushed to the time the i.v. was removed; number of i.vs. removed because of infiltration. RESULTS: No statistical or clinical differences between the three groups for duration of i.v. nor for incidence of complications. CONCLUSIONS: The use of heparin in i.v. lock flush solution did not affect the duration of i.v. locks nor the incidence of infiltration in neonates. PMID- 9743927 TI - Children and violence. PMID- 9743928 TI - [Times change, does glaucoma also change over time?]. PMID- 9743929 TI - [The glaucoma work group--an initiative of the Tubingen University Eye Clinic]. PMID- 9743930 TI - [Intraocular pressure self-monitoring in glaucoma patients]. PMID- 9743931 TI - [New drugs in glaucoma therapy]. PMID- 9743933 TI - [Long-term results of autologous conjunctiva-limbus transplantation in pterygium]. AB - BACKGROUND: Conjunctiva-limbus autografting is known as a safe surgical technique for the removal of pterygia, with a low rate of recurrence. However, the long term effect of this surgical maneuver is not clear. This study now investigates the long-term efficacy of conjunctiva-limbus autografts to prevent pterygium recurrence. METHODS: Conjunctiva-limbus transplants for primary (n = 62) or recurrent (n = 8) pterygia were reevaluated 11 to 83 months after surgery (mean: 44.97 months). Slit-lamp appearance and photomicrographs were studied with respect to the configuration of the transplant and the recurrence of the fibrovascular tissue typical of pterygia. RESULTS: Corneal pterygium recurrence has been observed in 2 cases. Fibrovascular tissue was found at the peripheral transplant-margin in 15 cases, and transplant compression towards the limbal margins were detected in further 7 patients. These conjunctival changes have not been observed during the first postoperative months. CONCLUSIONS: Conjunctiva limbus autografts in pterygia have excellent efficacy against recurrence within the first few years. The transplant compression and fibrovascular changes within the peripheral conjuctiva seen in this study suggest that recurrencies might, however, develop on the long-term. PMID- 9743932 TI - [Risk factors for trabeculectomy failure]. AB - BACKGROUND: Trabeculectomy is today the filtering procedure of choice, because complications are rare and success-rates high. A modification of our technique (fornix-based conjunctival flap closed by a running suture) introduced 3 years ago, has lead us to this retrospective assessment of complications and success rates. Some of the risk factors for failure are known, others however remain obscure. This retrospective series was also used to further identify some of these risk factors for trabeculectomy failure. PATIENTS AND METHODS: All 388 trabeculectomies performed between January 1992 and June 1994 at our hospital were included in the study. Important pre- and postoperative data were retrospectively assessed from patients case notes, with a special interest in the course of post-operative intra-ocular pressure (IOP). Two major groups were differentiated: Those with open-angle glaucoma (OAG) (i.e. primary open-angle glaucoma (POAG), pseudoexfoliation glaucoma (PEX) and pigment dispersion syndrome (PDS)), and other various glaucoma diagnoses. Risk factors were assessed using Cox-proportional hazard model adopting three different criteria for success. RESULTS: The best success-rate after 12 months of follow-up had patients with narrow angle glaucoma (93.1%)(at least those suitable for filtering surgery), followed by patients with POAG (92.8%), 2 degrees open-angle glaucoma (81.8%), aphakic (75%), juvenile (70.6%) and PDS (52.9%). Pseudophakia and development of an encapsulated bleb (Tenon' cyst) were identified as significant (p < 0.05) risk factors for failure. In addition, YAG-Laser Iridotomy in OAG-group and Aphakia in the group of various glaucoma diagnoses were identified as risk factors for successful post-operative IOP control. CONCLUSIONS: Filtering surgery (trabeculectomy) is a potent method to reduce IOP. Pseudophakia and an encapsulated bleb are the main risk factors for surgical failure. Because of amazingly high success-rates we tend to perform filtering surgery today earlier than ten years ago, especially as previous long-term topical antiglaucoma treatment may reduce filtering surgery success. PMID- 9743934 TI - [Protective ptosis by botulinum A toxin injection in corneal affectations]. AB - BACKGROUND: Botulinum toxin A has been introduced as a local injection therapy of different conditions with focal muscular hypercontractions. In the ophthalmologic field the toxin has proven its efficacy in the therapy of blepharospasm and hemifacial spasm. There are only few reports on the use of a botulinum toxin A to induce a protective ptosis in patients with persistent corneal ulcers. PATIENTS AND METHODS: 21 patients who failed to respond to conservative therapy of corneal erosions or ulcers of different origin received a botulinum toxin A injection into the levator palpebrae superioris muscle. RESULTS: The ptosis began after a mean of 1.5 days (1-3 days) and was complete after a mean of 5.1 days (3-12 days) after injection. Complete recovery of the levator function could be observed after a mean of 12.4 weeks (4-24 weeks). In 13 patients (61.8%) the botulinum toxin A induced protective ptosis lead to a complete healing of indolent ulcers or erosions, in 4 patients (19%) an additional tarsorrhaphy was necessary. In 3 patients no healing could be observed during follow up, in one patient (with neuroparalytic ulcer) the injection was given prophylactically. The period of healing on average was 3.8 weeks. There was no relationship between the healing rate and the duration of the corneal disease prior to the botulinum toxin injection. The mean healing rate of younger patients was higher (75%) than that of older patients (53.8%) and higher in erosions (70%) than in ulcers (30%). No side effects were observed besides in one patient the undesirable duration of the ptosis of a half year. CONCLUSION: The induction of a protective ptosis with botulinum toxin A injection is an efficacious treatment alternative in persistent corneal erosions and ulcers before performing a tarsorrhaphy. This method is preferrable especially in patients with lagophthalmos due to facial nerve paresis with potential recovery. PMID- 9743935 TI - [Autofluorescence characteristics of lipofuscin components in different forms of late senile macular degeneration]. AB - BACKGROUND: Lipofuscin is the main fluorophore of the human fundus. Because lipofuscin is the result of the accumulation of metabolic debris in pigmentepithelial cells (RPE), the autofluorescence can be interpreted as a clinical sign for the metabolic activity of the RPE. In order to get informations of RPE-function in different types of late AMD, the autofluorescence patterns in patients with late AMD were analyzed. MATERIAL AND METHOD: A prospective examination of the fundus-autofluorescence of 64 eyes of 52 patients with different types of late AMD was performed using a confocal scanning-laser opthalmoscope. The autofluorescence images were categorized in respect to the type of late AMD according to the opthalmoscopic and fluoresceine-angiographic findings. RESULTS: Reduced autofluorescence was found in the centre of occult (78.6%) and classic (100%) choroidal neovascularisations (NV) as well as in the occult NV of RPE detachments. A loss of autofluorescence was related to the RPE free area of RPE-tears (100%) and to RPE-atrophy (88.9%) with sometimes increased autofluorescence at the rim. Increased autofluorescence could be seen at the surface of RPE-detachments (71.4%), in the area of the shrink age of RPE in RPE tears (100%) as well as at RPE-proliferations in small occult NV (100%). Disciforme scars showed variable patterns of autofluorescence. CONCLUSION: The autofluorescence of the RPE can be analyzed clinically with the described method. Different patterns of autofluorescence could be revealed in different types of late AMD. Increased autofluorescence was found in lesions with proliferative or phagocytotic metabolic activity of the RPE like RPE-detachments, shrinked RPE in RPE-tears or occult NV with RPE-proliferations. The reduced autofluorescence in occult or classical choroidal NV can be interpreted as a sign of decompensation of the RPE and was also seen in areas with RPE-loss. PMID- 9743936 TI - [Dynamics of accumulation and degradation of lipofuscin in retinal pigment epithelium in senile macular degeneration]. AB - BACKGROUND: It is thought that lipofuscin plays a central role in the pathogenesis of age-related macular degeneration (AMD). The lack of histopathological material has been a severe limitation in our knowledge on lipofuscin in this disease. A new technique has been developed that allows in vivo imaging of fundus autofluorescence derived from lipofuscin in the retinal pigment epithelium (RPE) using a confocal Laser Scanning Ophthalmoscope (LSO). We studied the dynamics of lipofuscin accumulation and degradation in patients with AMD. MATERIALS AND METHODS: Serial examinations of the spatial distribution of fundus autofluorescence were performed in 148 eyes of 74 patients with AMD using a LSO over a period of 1-3.5 years. RESULTS: Fundus autofluorescence changed over time in almost all eyes studied. Areas of increased autofluorescence occurred progressively during follow up in eyes with drusen and hyperpigmentation. The size of pathologic autofluorescence increased over time in almost all eyes with geographic atrophy, subretinal neovascularisations and disciform scars. Irregular autofluorescence was seen over most subretinal neovascularisations. Autofluorescence intensity decreased in old subretinal neovascularisations and disciform scars over time. CONCLUSIONS: Changes of the distribution of autofluorescence occur in eyes with AMD over time. Fundus autofluorescence imaging allows in vivo analysis of the dynamics of accumulation and degradation of lipofuscin in the RPE in eyes with AMD and documentation of metabolic activity of the RPE. PMID- 9743937 TI - [Keratinocyte density of the cornea in vivo. Automated measurement with a modified confocal microscopy MICROPHTHAL]. AB - BACKGROUND: The MICROPHTHAL is a confocal slit light scanning microscope for a non-invasive in-vivo examination of corneal structures of human eyes. With this instrument even thin layers of corneal tissue can be imaged in good quality. Otherwise, blurring of single frames and deviations from the z-axis in video sequences caused by high speed movements of the eye would normally prevent a measurement the density of keratocytes in the cornea. The goal of the investigation was optical pachymetry, the automatical measurement of the keratocytes density and a 3D-dimensional reconstruction of the central cornea in vivo under constant imaging conditions. MATERIALS AND METHODS: We developed a low vacuum suction cup system for stabilizing the eye in front of the microscope objective during the z-scan through the cornea. A stepmotor shifting system for the objective locates inside the suction cup with a central hole was installed underneath them icroscope. Control of this system via computer facilitated shifting the focal plane along the z-axis. The layer images were recorded using a S-VHS-tape and saved on the PC. The digital analysis was performed using a special software to automatically and off-line evaluate the density of keratocytes in combination with the 3D-reconstruction. The software also corrected the background illumination and small axial jitter. After this procedure the keratocytes density and the 3D-reconstruction in 70 images of the z scan were calculated. We examined 47 corneas of 25 healthy probands. The range of age was 25-56 years. Independent control evaluation of the video sequences were taken manually on an INDIGO HIGH IMPACT workstation. RESULTS: By assign all keratocytes to the corneal measurement volume we found a averaged density of 15,730 cells/mm3 in the central cornea. The averaged thickness of the cornea was 0.556 mm. The control valuation of identical video-sequences on the workstation accomplished the same result of 16,000 keratocytes/mm3, also similar the result of the automatically measurement with the modified software. CONCLUSIONS: This modification of the microscope is a promising in-vivo tool for optical pachymetry and quantitative examination of corneal microstructures. The stabilization effect of the low-vacuum suction cup system in the front of the microscope for computer controlled valuation of the density profile of keratocytes and the 3D reconstruction of a central corneal volume element has produced encouraging results. Characterization of pathophysiological changes in the distribution of keratocytes after excimer laser ablation for phototherapeutic or photorefractive keratectomy, for example, can be estimated without pain for the patients. PMID- 9743938 TI - [Holzwig iris retractor and use in narrow pupils]. AB - BACKGROUND: The objective was to develop an easy to handle tool for temporary implantation to dilate narrow pupils. MATERIAL AND METHODS: High molecular PMMA was used to lathe cut a v-shaped device with sides of 6.9 mm length, 0.8 mm height and 0.4 mm width which opens 7.0 mm. Positioning holes of 0.5 mm diameter are located in the caudal ends of the sides. The HIR was used during cataract extraction in 24 patients with mydriatic-resistant pupils, amblyopia or distinctive macula degeneration. RESULTS: In all 26 cases cataract extraction was made easier due to physiologically dilated pupils. The risk of an intraoperative miosis was effectively eliminated. In two cases removal was difficult as the lower tunnel lip interfered with the removal of the HIR. CONCLUSION: In comparison with the commercially available devices, the HIR is the simplest, safest and most physiologic alternative. PMID- 9743939 TI - [Topical administration of metronidazole gel as an effective therapy alternative in chronic Demodex blepharitis--a case report]. AB - BACKGROUND: Blepharoconjunctivitis is the commonest manifestation of ocular rosacea. Cilia epilation proves Demodex folliculorum, considered an etiologic factor in rosacea. Complications and differential diagnosis include dry eyes, seborrheic, bacterial and allergic blepharoconjunctivitis. Treatment involves lid scrubs and mercury ointment, its duration is limited to 6 weeks under frequent control due to corneal toxicity of mercury. HISTORY AND SIGNS: 30-year-old female with complaint of red, irritated eyes for 21 years, resistant to antibiotics and antiallergics. General medical history unremarkable, mercury allergy. Acuity: R/L 20/20. Biomicroscopy: red, thickened eyelid margins, crusty debris on rarefied, partially broken lashes, conjunctival telangiectasia, low tear meniscus, further ophthalmologic examination unremarkable. DIAGNOSIS: chronic Demodex blepharoconjunctivitis. THERAPY AND OUTCOME: Conventional treatment was contraindicated due to mercury allergy. Alternative oral minocycline 100 mg daily brought no subjective nor objective relief. Combination of lid scrubs and 2% Metronidazole gel relieved symptoms and halved number of mites after 1 month, lashes grew again after 2 months. Treatment was discontinued after 6 months as Demodex folliculorum proof remained negative. No relapse occurred during 1 year. CONCLUSIONS: Topical Metronidazole 2% proved to be an effective treatment of chronic Demodex blepharoconjunctivitis in our case and thus may offer a new and save alternative to existing therapies. Neither ocular nor systemic side effects occurred. Controversial theories on the aetiology of blepharitis are discussed. PMID- 9743940 TI - [Choroid infiltration in myelodysplastic syndrome]. AB - BACKGROUND: Myelodysplastic syndrome is a clonal disease of the hematopoetic system characterized by insufficiency of affected bone marrow cell lines. After long-term course of myelodysplastic syndrome an acceleration towards acute myeloic leukemia is a frequent finding. Involvement of the eye is a well known phenomenon in acute leukemia or in blast crisis with chronic leukemias. Eye involvement in myelodysplastic syndrome showing its transition into acute myeloic leukemia however has been published in only few cases. CASE REPORT: We present a 70-year-old male patient suffering from myelodysplastic syndrome, complaining of an acute visual decrease to 0.05 in the left eye. Clinical findings, ultrasound and fluoresceine angiography were in accordance with choroidal infiltration. From the hematologic findings, the myelodysplastic syndrome had been in partial remission after chemotherapy without any sign of relapse or exacerbation. Only because of the ophthalmologic diagnosis, bone marrow aspiration was performed and revealed progression of myelodysplastic syndrome to acute myeloic leukemia. Prompt administration of chemotherapy and external radiation of the posterior pol of the eye led to complete resolution of the fundus lesion within 10 days and visual acuity recovered to 0.8. CONCLUSION: To the best of our knowledge this is the first patient with a choroidal infiltration as the initial sign of progression of myelodysplastic syndrome to acute myeloic leukemia. Realizing this possibility helps for an early diagnosis and rapid therapy which is so crucial in prolonging life. PMID- 9743941 TI - [Prevention of amblyopia in acute eyelid closure--a new method for keeping the optical axis open by insertion of a small tube]. AB - BACKGROUND: Complete eyelid closure by capillary eyelid hemangioma or ptosis in the first months of life is an indication for acute measures to prevent amblyopia. Since it is sometimes not possible to hold up the affected upper eyelids with adhesive tape (mechanical obstacle, danger of skin maceration in involvement of the forehead) or not sufficiently (intense divergence), an alternative method will be presented for emergency treatment of blockade of the optical axis. METHODS: The eyelids are kept open mechanically with a cylindrical tube of perspex (scleral immersion shell), which is normally used for echography of the anterior segment and for biometry (immersion technique). After surface anesthesia, it can be readily inserted. Its area of contact to the sclera has the form of a scleral shell. A drop of lubricant is applied into the tube at intervals of about five minutes. PATIENTS AND RESULTS: In a six-week-old girl with complete eyelid closure owing to a facial hemangioma, adequate eyelid opening could only be achieved by insertion of the scleral immersion shell. In an eleven-months-old boy with complete ptosis and divergence as well as vertical deviation, the optical axis could only be kept open by insertion of the scleral immersion shell and by simultaneous displacement to the temporal side (adhesive tape). The uncomplicated performance for up to two hours daily was initially carried out in the hospital and later by the parents, and could be terminated after five and three and a half weeks, respectively, thanks to improvement due to therapy or spontaneous improvement. CONCLUSIONS: Mechanical eyelid opening by insertion of a scleral immersion shell serves to bridge over the time interval to the onset of spontaneous improvement or the success of a causal therapy. The advantages consist in the good handling, also for parents, the low danger of injury and the ubiquitous and rapid availability (basic equipment of an ophthalmological ultrasonography unit). PMID- 9743942 TI - [Ophthalmomyiasis externa caused by Oestrus ovis in Franconia]. AB - BACKGROUND: Ophthalmological diseases caused by parasites are a very rare entity in middle europe. Ophthalmomyiasis caused by parasites living in our country are only reported as case history in literature. PATIENT: We report on a 28-year old patient, seen in our department in august 1997 suffering from ophthalmomyiasis externa. CASE HISTORY: The male patient was riding on a motorbike when a fly gets into his right eye. 12 hours later he had a feeling of pain in the eye. When having a closer look to his right eye, he found some fly larva (2.0! 0.8 mm) in the conjunctival sack. The patient could extract the fly larva from the conjunctival sack by himself. On examination at our department we found a unspecific irritation of the conjunctiva. The fly larva was identified as larva of oestrus ovis ("Schafsbremse"). CONCLUSION: Although ophthalmomyiasis is a very rare entity in middle europe single appearance is possible even in our country. No specific therapy was necessary in this case. PMID- 9743943 TI - [Latanoprost (Xalatan)-induced macular edema]. AB - BACKGROUND: Latanoprost represents a new therapeutic option in the treatment of chronic open angle glaucoma. It has only recently been reported for the first time that latanoprost caused cystoid macular edema in pseudophakic patients. CASE REPORT: A 60-year-old pseudophakic patient who suffered from a 10-year history of glaucoma, revealed a cystoid macular edema on fluorescence angiography after 10 days of treatment with latanoprost. One week after cessation of latanoprost therapy the cystoid edema in fluorescence angiography had resolved and the vision improved from 0.4 to 0.8. CONCLUSIONS: With respect to this severe complication, latanoprost should be used with great care and in clear indications, particularly in patients with risk of blood-aqueous leakage e.g. pseudophakic patients. PMID- 9743944 TI - Molecular determinants of apoptosis induced by cytotoxic drugs. AB - Recent experimental evidence suggests that apoptosis pathways such as the CD95 system are an important mediator of chemotherapy-induced apoptosis in various tumor cell lines. Therapeutic concentrations of cytotoxic drugs induce CD95 and CD95-L that mediates apoptosis via an autocrine/paracrine loop by crosslinking CD95. Interfering with CD95-L/receptor interaction by antagonistic antibodies to the receptor or by inhibition of CD95-L expression strongly reduces apoptosis. Drug-induced apoptosis critically depends on activation of caspases since apoptosis is almost completely abrogated by the caspase inhibitor zVAD-fmk. The receptor apical caspase FLICE/MACH (caspase-8) and the downstream caspase CPP32 (caspase-3) are cleaved resulting in processing of substrates such as the nuclear enzyme PARP. In addition, the response to cytotoxic drugs is modulated by pro- and antiapoptotic proteins of the Bcl-2 family and p53. Defects in apoptosis pathways, e.g. deficient upregulation of CD95-L, downregulation of CD95 expression or blockade of caspase activation may confer resistance to cytotoxic drug treatment. Thus, chemosensitivity of tumor cells depends on intact apoptosis pathways such as the CD95 system that are activated by chemotherapeutic drugs. These findings may have implications for drug sensitivity and resistance of tumor cells. PMID- 9743945 TI - [The role of CD95 (APO-1/Fas) mutations in lymphoproliferative and malignant lymphatic diseases]. AB - CD95 (APO-1/Fas)-mediated apoptosis plays a major role in normal lymphocyte regulation. CD95 mutations cause a benign autoimmune lymphoproliferation syndrome (ALPS) in mice and humans. CD95 is mutated in some de novo T-lineage acute lymphoblastic leukemia of childhood and these mutations might be of biological significance. The resistance toward CD95-mediated apoptosis observed in most B lineage ALL is not caused by mutations of CD95. CD95 mutations have been associated with Hodgkin's and Non-Hodgkin's lymphoma and have been described in multiple myeloma. The relevance of CD95 mutations for chemoresistance of ALL requires further study. PMID- 9743946 TI - [Pharmacokinetic aspects of oral administration of etoposide]. AB - BACKGROUND: Etoposide is a cytotoxic agent which is frequently employed in paediatric oncology and which is available for intravenous as well as oral application. Many advantages of the formulation for oral use have been opposed by concerns about its interindividually varying bioavailability. The influence of the dosage of etoposide on its activity and toxicity ("schedule dependency") has also been discussed. The present paper deals with the pharmacokinetics of oral etoposide focusing on the interindividual variability. PATIENTS: Sixteen patients aged between 3 and 73 years received oral etoposide at a dosage of 28 mg/m2 to 149 mg/m2 in combination with oral trofosfamide for palliation. METHOD: HPLC was used to measure total and free serum etoposide in 16 patients, and the etoposide concentration in several urine samples from 8 patients. Pharmacokinetic parameters were normalized to a dosage of 100 mg/m2. RESULTS: The peak serum concentration, the time to peak concentration, the area under the concentration time curve, the terminal half-life and the apparent clearance after oral application were calculated to be 6.7 micrograms/ml, 2.1 h, 51.8 (microgram.h/ml)/(100 mg/m2), 5.6 h, and 40.3 ml/min for total etoposide, and 0.23 microgram/ml, 1.9 h, 1.76 (microgram.h/ml)/ (100 mg/m2), 5.9 h, and 1172 ml/min for free serum etoposide, respectively. On an average, urinary recovery of etoposide was 21% of the oral dose. The fraction of free etoposide was calculated to be close to 4%. Regarding the systemic exposure to etoposide, a variation coefficient of 40% was determined. Additional studies showed that the interindividual variability mainly concerned the peak levels, while the duration for which intermediate etoposide levels were maintained varied less between individuals. On simulating different dosage schedules, it was seen that the duration of intermediate concentrations (0.5-2 micrograms/ml) may be extended significantly by dividing the daily dose of etoposide into two oral applications. CONCLUSION: The total systemic exposure under oral etoposide treatment varies considerably between individuals. Extended intervals of intermediate etoposide concentration and less variation are, however, possible with oral therapy. Dividing the daily dose into two applications seems advisable. Future studies are warranted to test hypotheses on pharmacokinetic-pharmacodynamic aspects by pharmacological drug monitoring. PMID- 9743948 TI - Defunct hematopoietic progenitor growth and heterogeneous immunological phenotypes in acquired aplastic anemia of childhood: identification of subsets with decreased hematopoietic progenitors and increased IL15 or IL10 production. AB - Acquired aplastic anemia (AAA) is a disorder with multiple causes. The pathogenetic mechanisms leading to marrow failure are diverse. The clinical finding, that most patients respond to immunosuppressive therapy implicates an immune pathophysiology of AAA. On the other hand successful and long lasting reconstitution after allogeneic stem cell transplantation implicates, that a stem cell defect is a major pathogenetic factor. We analyzed immunological and hematological parameters on 91 pediatric patients with AAA at time of diagnosis. We defined three distinct pathogenetic subgroups by analyzing the frequency of hematopoietic progenitors and the frequency of activated T-cells in patients bone marrow: type I: decreased percentage of hematopoietic progenitors; type II: increased percentage of activated T-cells; type III: increased percentage of pluripotent progenitors. In 51% of the AAA patients we found a relative decreased frequency of hematopoietic progenitors and 29% of the patients demonstrated an increased percentage of activated T-cells, 25% patients showed an increased percentage of pluripotent hematopoietic progenitors. In order to characterize these distinct immunological subgroups, we investigated the plasma levels of IFN gamma and IL15 as inhibitors and IL10 as stimulator of hematopoiesis. IL15 plasma levels were significantly elevated in AAA patients when compared to controls. In contrast we could not demonstrate a difference between IFN-gamma or IL10 plasma levels in AAA patients when compared to controls. However for IL10 and IL15 we were able to define subgroups of patients with highly elevated plasma levels of the cytokines. These data indicate that the immunopathogenesis of AAA can be heterogeneous. This heterogeneity might reflect exposure to different exogenous agents or heterogeneity in intrinsic susceptibility. The clinical impact of our findings needs to be assessed in long-term follow up of the patients. PMID- 9743949 TI - Relapse and clonal disease in children with aplastic anemia (AA) after immunosuppressive therapy (IST): the SAA 94 experience. German/Austrian Pediatric Aplastic Anemia Working Group. AB - Since the introduction of combined immunosuppressive therapy (IST) into management of aplastic anemia (AA) in childhood response and probability of survival improved. In contrast to bone marrow transplantation (BMT), however, patients after IST are not considered cured as high rates of relapse and development of clonal disease demonstrate. From 11/93 to 9/97 114 children (65 m, 49 f; median age 9.5 y.) from 37 centers in Germany and Austria were registered in the SAA 94 study. 86 patients lacking a matched sibling donor received IST. Most of the patients suffered from very severe (VSAA: PMN < 200/microliter) or severe AA (SAA: PMN < 500/microliter). All patients were treated with combined IST consisting of ALG and Cyclosporin A (CSA). VSAA and SAA patients were additionally treated with G-CSF. Therapy response was evaluated at day 112, after 6, 12 and 18 months. 8/86 patients died, the probability of survival being 87% after 4 years. At d 112 61% of evaluable patients became independent of transfusions (IST response: CR + PR), 13% with normal blood counts (CR). After 6 months 33% showed CR. At 12 and 18 months response improved to 74% resp. 80%, 39% resp. 55% CR. The best response was achieved in the subgroup of VSAA with 90% (PR + CR) and 65% CR after 18 months. 4 patients developed AML 3-19 months after the beginning of IST. In 2/4 pts. an aberrant clone (-7; 5q-) could be detected retrospectively in BM at diagnosis of AA. 3 nonresponders developed chromosomal aberrations (+19; -7, +12; +8) after 4, 12 and 16 months without morphological signs of AML or MDS. Overall 11 relapses occurred at a median time of 12 months (range 5-27 months) after the beginning of IST. 2 of them relapsed under CSA therapy, 2 under tapering of CSA and 7 after cessation of CSA. 7 patients responded again to CSA monotherapy. Overall response rate is 77% with a probability of event free survival (EFS) of 54% after 4 years regarding all complications mentioned as events. PMID- 9743950 TI - [Treatment of hemophagocytic lymphohistiocytosis, HLH, with bone marrow transplantation]. AB - Hemophagocytic lymphohistiocytosis (HLH) is a rare disease of infancy and young childhood. The clinical presentation includes recurrent unexplained fever with hepatosplenomegaly. Cytopenia, hypofibrinogenemia and/or hypertriglyceridemia and hemophagocytosis in bone marrow, spleen and lymphnode confirm the diagnosis. Hemophagocytosis may not be present at the beginning. In these cases, diagnosis is facilitated by a positive family history, a relapsing course of the disease, the frequent involvement of the central nervous system and positive findings on immunological work-up. Treatment by chemotherapy and immunosuppressants can achieve sustained remissions in most patients and reinduction of remission after relapse is possible. Most children however, eventually die from progressive disease. At present, allogeneic bone marrow transplantation is the only curative therapeutic option. Between August 1992 and May 1997 eleven consecutive patients with HLH received bone marrow from unrelated (n = 7) or matched sibling donors (n = 4). The conditioning regimen consisted of busulfan, VP-16 and cyclophosphamide. Patients engrafted after a median time of 16 days (13-43). Only one patient developed grade III acute GVHD, another patient, grade II acute GVHD. Although regimen-related toxicity was extensive, all patients have survived without signs of HLH after a median follow up of 20 months (8-63). One patient suffers from chronic GVHD, three patients reveal psychomotoric retardation and one patient has severe impairment with spastic tetraparesis, amaurosis and seizures. Our experience shows that HLH can be successfully treated by allogeneic BMT from unrelated donors. PMID- 9743951 TI - German case control study on childhood leukaemia--basic considerations, methodology and summary of the results. AB - In order to explore potential risk factors of childhood leukaemia, a case control study was performed including all incident cases from 1992 to 1994. The study was based on the German Childhood Cancer Registry. It was restricted to cases from West Germany and extended retrospectively until 1980 for children who were living in regions covered by a previous incidence study on nuclear installations (21). The study was conducted in close correspondence with a preceding case control study in Lower Saxony (13). Results of this study and of others published in the literature were used to define explicit hypotheses for the present study. This paper presents the methodology of the study and gives an overview of some basic results. More detailed analyses of the investigated potential risk factors will be published elsewhere. The study comprised a total of 2358 cases (leukaemias, lymphomas, selected tumours) and 2588 controls. Response rates were 81% for cases and 67% for controls. For leukaemias, the main results regarding maternal factors, pregnancy, birth, immune system, ionising radiation, parental occupation and environmental factors were as follows: Positive associations were observed between childhood leukaemias and young maternal age at birth, high birth weight, tonsillectomy and use of pesticides. Some results suggest a protective effect for allergies and vaccinations. A negative association was observed with maternal smoking and childhood leukaemia. No associations were found with frequency of stillbirths, maternal alcohol consumption, parental exposure to benzene and use of wood preservatives. X-ray examinations in early childhood and parental radiation exposure did not show any consistent associations with leukaemia. Potential risk factors were not reported more frequently by cases and controls living in 114 communities with increased incidence rates. The strength of our study lies in the large number of participating families and in the population based approach. PMID- 9743952 TI - Prevention of CNS recurrence in childhood ALL: results with reduced radiotherapy combined with CNS-directed chemotherapy in four consecutive ALL-BFM trials. AB - BACKGROUND: The introduction of cranial radiotherapy (CRT) has provided efficient control of overt or subclinical meningeosis in acute lymphoblastic leukemia (ALL). Especially due to the long-term toxicity of CRT, reduction or elimination of radiotherapy appeared mandatory after cure rates of more than 70% had been achieved in ALL. The Berlin-Frankfurt-Munster (BFM) Study Group initiated several attempts in certain ALL subgroups to omit or reduce CRT while using more CNS directed chemotherapy but without extended intrathecal treatment during maintenance therapy. This analysis summarizes the essential results that are in particular relevant because irradiation of the central nervous system (CNS) has been further reduced in the most recent trial ALL-BFM 95. PATIENTS AND METHODS: More than 4000 patients enrolled between 1981 and 1995 in one of the last four ALL-BFM trials have been analyzed to demonstrate the efficiency of intensive systemic and intrathecal chemotherapy with or without reduced CRT in the prevention of CNS relapses. RESULTS: In trial ALL-BFM81, it was shown that only in low-risk (LR) patients preventive radiotherapy can be replaced safely by intermediate dose (0.5 g/m2) methotrexate (MHD-MTX). In intermediate risk (IR) patients this attempt failed: IR pts had 8 times more CNS relapses if treated by MHD-MTX without CRT. In the subsequent trial ALL-BFM 83, all pts received MHD MTX. IR pts were randomly treated with 12 or 18 Gy of preventive CRT which did not result in a significantly different outcome. The results from the subsequent trial ALL-BFM 86, using a more intensive consolidation with high-dose methotrexate (HD-MTX), demonstrated that the elimination of CRT in low-risk ALL, the reduced CRT of 12 Gy for IR, 18 Gy for medium (MR), and the reduced CRT with 18 Gy for high risk (HR) ALL, respectively, was justified: the incidence of relapses with CNS involvement was reduced to less than 5% (Reiter et al. 1994, Blood 84: 3122). When even less intensive preventive CRT (12 Gy for all medium and high risk patients) was used in trial ALL-BFM 90, the rate of CNS-related relapses was again below 5%. HR patients now treated with more CNS-directed chemotherapy had the lowest rate of CNS-related relapses observed so far in the BFM trials, even though CRT was also reduced to 12 Gy. Patients with T-cell ALL were shown to be protected from CNS recurrence by the combination of CRT (12 Gy) and HD-MTX more effectively than by HD-MTX in consolidation and TIT therapy during maintenance, especially if they presented with high WBC as shown in a joint AIEOP/BFM analysis (Conter et al. 1997, JCO 15: 2786). Patients with overt meningeosis which are characterized by a high leukemic cell load at diagnosis had a rate of recurrences that was comparable to that of patients with high WBC but no CNS disease. CONCLUSION: Low-risk ALL patients can be efficiently prevented from CNS relapse by intensive systemic and intrathecal chemotherapy without CRT. Patients with intermediate or medium risk ALL, including T-cell ALL, did not suffer from more CNS or systemic relapses when CRT was reduced to only 12 Gy. Patients with inadequate response to therapy are at particularly high risk for relapse with CNS involvement. Therefore, more CNS-directed systemic and intrathecal chemotherapy was applied in trial ALL-BFM 90, combined with only 12 Gy cranial irradiation, and improved the control of CNS recurrence. It seems likely that larger subsets of B-precursor ALL can be protected from CNS-related relapse by intensive chemotherapy without extended IT treatment and without CRT. This is being investigated in the ongoing trial ALL-BFM 95. PMID- 9743953 TI - [Late sequelae of CNS recurrence of acute lymphoblastic leukemia in childhood]. AB - Study objective was to evaluate retrospectively central nervous system (CNS) morbidity of children with acute lymphoblastic leukemia treated with intensive chemotherapy and cranial radiotherapy for a first isolated or combined CNS relapse. Neurological (Touwen), neuropsychological (CFT 20, Wechsler scales, d2 attention test) and neuromorphological (CT, MRI) assessments were performed in 17 children (9 girls, 8 boys) aged between 7 and 14 years. Patients were off therapy for median 4 years; cranial radiotherapy for CNS relapse (12-24 Gy) was given to all patients 2 to 9 years ago (median 5.5 years). Ten patients had received preventive cranial radiotherapy during front-line treatment, previously. In this group, the cumulative radiation dose ranged between 30 and 39 Gy. Patients received 12 to 30 intrathecal methotrexate doses (median 22). Compared with normative levels for age (100 points) performance IQ (89.9) and full scale IQ (92.0), Culture Fair IQ (88.3) and attention and concentration (90.9) were significantly impaired. Verbal IQ (95.5) was not significantly different from normal expectations. Neurological investigations of 16 patients showed mild signs in 7 (44%) of them, mostly of fine-motor skills and coordination, but no major motor disability. In MRI and CT, white matter changes or ventricular enlargement were noted in 10 of 16 patients (63%). Parent-completed questionnaires indicated problem behaviors in 5 children. Cumulative doses of methotrexate correlated significantly with neuropsychological test results. Children irradiated twice as well as girls had poorer cognitive functions, not being statistically significant. Despite pathological test results, all children attended a normal school and did not exhibit obvious impairment in daily life. In conclusion, CNS relapse and attendant therapy, mostly consisting of a second course of cranial radiotherapy, caused measurable intellectual deficits and CNS morbidity, which did not result in severely impaired performance. Periodic psychological und clinical examinations are recommended for recognition of delayed morbiditiy and early intervention. PMID- 9743954 TI - [Prognosis of children with acute myelocytic leukemia after first relapse]. AB - Nearly 40% of children treated within the AML BFM studies experience recurrence of their disease after having achieved remission. In our retrospective analysis we tried to estimate prognosis after relapse in children treated with intensive relapse regimens and studied the impact of prognostic factors for second remission and survival. PATIENTS: 102 patients suffering from first relapse were treated intensively according to the relapse protocols BFM REZ91 and REZ93 or intensive salvage therapy consisting of double induction with high dose Ara-C, mitoxantrone and VP-16. Once in CR, patients continued to receive a 6-week consolidation and either allogeneic or autologous bone marrow transplantation (BMT). RESULTS: Time to relapse was in median 1.1 years, range 0-8 years. Fifty two of 102 pts. (51%) achieved 2nd remission (CR), 10 (10%) partial remission, 37 (36%) were nonresponders, and 3(3%) died early during salvage therapy. Twenty seven were still in CR, median 2.5 years, range 0.4-7.0 years, with an overall survival of 21%, SE 5% after 5 years. The response and survival rate was similar in all treatment groups. Fifty patients were transplanted, 43 being in 2nd CR, and 7 with residual blasts. Twenty-seven patients received an allograft: Twenty one from a matched sibling (MSD), 1 from a haploid and 5 from a matched unrelated donor (MUD); 23 received an autograft. None of the patients transplanted in partial remission survived. Whereas 7 of 16 patients were alive after MSD in 2nd CR, 1 after haploid BMT. Four of 5 patients died after MUD BMT. Multivariate risk factor analysis revealed duration until relapse to be the most important factor for survival after relapse. The maximum risk-ratio was obtained at a threshold value of 1.5 years after diagnosis resulting in a 5-year survival of 10%, SE 5% for early relapse, and 40%, SE 10% for late relapse, p logrank 0.0001. CONCLUSION: Intensive relapse regimens can induce a 2nd CR in half of the patients. Children with late relapse (> 1.5 years after diagnosis) have a realistic chance for longtime survival. PMID- 9743956 TI - Advances in treatment techniques and time/dose schedules in external radiation therapy of brain tumours in childhood. AB - Recent advances in radiotherapy techniques seek to improve the therapeutic ratio in childhood brain tumours by adding potentially more effective treatment in ways that will increase tumour control and limit radiation toxicity. Stereotactic irradiation techniques in conjunction with rigid head fixation systems comprising single high-dose delivery ("radiosurgery"), fractionated convergence therapy and three dimensional conformal therapy focus the dose to tumour while sparing surrounding normal tissue. Although these techniques are well established in adults data for childhood central nervous system malignancies are scarce. Preliminary data reveal low acute toxicity and promising results in recurrent tumours as well as in primary treatment. Whether stereotactic radiation therapy with either of the latter techniques will add substantially to disease control and preserve neurologic function remains to be established and should be part of future investigations. The introduction of quality control procedures assures precise radiotherapy of the tumour site, whole brain and craniospinal axis and is a prerequisite to improve survival and to reduce potential adverse effects. Hyperfractionated radiotherapy has the potential of increasing dose to tumour more safely theoretically sparing children some of the late effects while in particular in craniospinal irradiation. Pilot studies revealed excellent tumour control in medulloblastoma with acceptable acute toxicity. The strategy is part of the subsequent HIT '98 trial both for low and high risk patients. In conclusion, there is considerable enthusiasm among all those caring for children with brain tumours to study modifications of radiation delivery that may result in improved treatment. Rapid accrual to an increasing number of cooperative prospective trials is expected to provide the body of data supporting the selection of novel radiation therapy modalities in the future. PMID- 9743955 TI - [Multi-national therapy study for Hodgkin's disease in children and adolescents GPOH-DH 95. Interim report after 2 1/2 years]. AB - Based on concepts of the successful German-Austrian pediatric Hodgkin studies DAL HD 78 until-90, a new trial was initiated addressing the question whether radiotherapy can be further reduced or can be omitted in case of complete remission after initial chemotherapy, aiming at reduction of sequelae after radiotherapy, especially radiogenic second malignancies. In respect to CHEMOTHERAPY patients are stratified into 3 therapy groups (TG) according to stage and gender: 2 courses of OPPA (girls) or OEPA (boys) in TG1 (stage IA/B, IIA), and in addition 2 (TG2: stage IEA/B, IIEA, IIB, IIIA) or 4 (TG3: stage IIEB, IIIEA/B, IIIB, IVA/B) COPP courses. Boys with stage IIIB and IIIEB receive OPPA instead of OEPA. RADIOTHERAPY is administered according to response to chemotherapy independent of stage: patients with complete remission or minimal residues do not receive irradiation; patients with more than 75% tumor regression are irradiated to involved fields at a dose of 20 Gy. Doses of 30 or 35 Gy are given to regions with tumor regression below 75% or residual bulky tumor of > 50 ml, respectively. INTERIM RESULTS: From 8/95 till 1/98 we registered 385 patients under the age of 18 years from Germany, Austria, Switzerland, Sweden and the Netherlands. Therapy has been completed in 334 patients. Three patients with solitary nodular paragranuloma were treated with surgery only. Out of 331 patients 89 (26.9%) achieved a complete remission with chemotherapy. Tumor regression of more than 75% was seen in 193 (58.3%) patients and below 75% in 39 (11.8%) patients. Tumor progression during chemotherapy occurred in 1 (0.3%) patient. Response after chemotherapy was not evaluable for 9 (2.7%) patients. Radiotherapy was omitted in 91 (27.1%) patients: in TG1 50 of 142 (34%) patients, TG2 24 of 98 (24.5%) patients and TG3 18 of 94 (19.2%) patients. Initially involved regions were irradiated at a dose of 20 Gy in 164 of 334 (49.1%) patients. Doses up to 30 Gy or 35 Gy were given to 19 (5.7%) or 57 (17.1%) patients respectively. Events (tumor progression, relapse or death) occurred in 23 of 334 patients until now. The event-free survival rate is 0.91 at 2 1/2 years for all study patients and 0.89 for patients without radiotherapy. Six relapses occurred in 91 patients without radiotherapy. No relapse occurred in TG1 (n = 49), but in 5 of 24 TG2-patients, and in 1 of 18 TG3 patients without radiotherapy. As yet, the results are not significantly inferior compared with trial DAL-HD 82. Therefore this trial aiming at omitting radiation therapy in patients with complete remission after a short lasting chemotherapy will be continued. Longer follow up is necessary for final evaluations and conclusions. PMID- 9743957 TI - Preradiation chemotherapy of children and young adults with malignant brain tumors: results of the German pilot trial HIT'88/'89. AB - BACKGROUND: Preradiation chemotherapy could be beneficial in malignant brain tumors, because the blood-brain tumor-barrier is disrupted after surgery, bone marrow recovery--essential for intense chemotherapy--is still intact, and CNS toxicity and ototoxicity of active drugs are lower before irradiation of a child's brain. PATIENTS AND METHODS: A neoadjuvant phase 2 and a single arm pilot trial were initiated to investigate the efficacy and toxicity of an intense multidrug regimen before radiotherapy in 147 patients aged between 3 and 29; 9 years with medulloblastoma (94), malignant glioma (22), ependymoma (21), and stPNET (10). They were treated with one or two cycles consisting of procarbazine, ifosfamide/mesna with etoposide, high dose methotrexate/CF, and cisplatin with cytarabine. RESULTS: Radiation therapy was delayed for 17-30 weeks (median 23 weeks) in 112 patients who received two cycles. Chemotherapy was well tolerated. Serious infections were observed in 20 patients, with one fatal fungal septicemia. In 69 high risk patients with a residual tumor and/or solid CNS metastases an objective response (CR plus PR) was achieved in 67% medulloblastoma, 57% stPNET, 55% anaplastic ependymoma and 25% malignant glioma. Progression-free survival (PFS) at 5 years was 57% in 14 high risk patients with medulloblastoma, who achieved a complete response (CR). After a less than CR the PFS was 20% (p = 0.01). Overall survival at 5 years was 57% in medulloblastoma, 62% in ependymoma, 36% in malignant glioma and 30% in stPNET. CONCLUSION: The HIT'88/'89 regimen was well tolerated and efficacious in regard to response rates and early PSF particularly in medulloblastoma and anaplastic ependymoma. Based on these results the prospectively randomized trial HIT'91 was designed to investigate the optimal timing of chemotherapy. Preradiation chemotherapy according to the HIT'88/'89 regimen was compared with the standard regimen using CCNU, cisplatin, and vincristine after radiation therapy. Additionally, strict quality control of the three treatment modalities was instituted to help improve the survival rates in both trial arms. PMID- 9743958 TI - Medulloblastoma: experience of a single institution. AB - BACKGROUND: The treatment of medulloblastoma has changed considerably during the last decades. Treatment differences between centers may affect a multicenter analysis. We analyzed data from patients of a single institution gathered over a long period of time. PATIENTS: Between 1968 and 1995, 60 patients with medulloblastoma were treated at the University of Munster. Thirty-six were male, 24-female. The ages ranged between 11 months and 32 years. METHODS: Data were retrospectively analyzed from files. Survival was estimated using the Kaplan Meier method and compared using the logranktest and multivariance analysis. RESULTS: The 5-year survival rate was 37%. This included an early mortality of 20% within the first two months, prior to 1980. Significant single, positive, prognostic factors included: no solid metastases (p = 0.001), age > 10 years (p < 0.002); total resection (p < 0.025); posterior fossa radiation with more than 50 Gy (p = 0.04); and intense chemotherapy (p = 0.02). Male patients did slightly worse (not significant). The three-year event-free survival rate of 16 patients treated after 1991 was 70%. CONCLUSION: The prognosis of medulloblastoma has clearly improved with the reduction of the perioperative mortality, standardized radiotherapy, and the introduction of intense chemotherapy. PMID- 9743959 TI - Monitoring tumor activity in low grade glioma of childhood. AB - Chemotherapeutic or radiotherapeutic regimens are being increasingly used in low grade glioma of childhood. These protocols require methods to monitor tumor activity. We report our experience in eleven patients. The tumors were localized in the optic pathway (3), cerebral cortex (4) and thalamus/hypothalamus (4). Histological diagnoses included low grade astrocytoma (6), gliofibroma (1) and ganglioglioma (2). Two children with neurofibromatosis type 1 (NF-1) and typical optical tumors were not biopsied. 13 episodes of progression were noted including 3 altered diagnoses. This was evident from clinical symptoms in 11/13 episodes, computed tomography (CT) or magnetic resonance imaging (MRI) in 10/13 situations, iodine-123-alpha-methyltyrosine (123I-IMT) single-photon emission computed tomography (SPECT) in 10/10 situations, fluorine-18 fluorodesoxyglucose (18F-FDG) positron emission tomography (PET) in 0/3 and thallium-201 (201Tl) SPECT in 1/1. Seven responses to chemotherapy were recorded. Clinical symptoms indicated this in 7/7 situations, MRI in 5/7, 123I-IMT SPECT in 1/2 and 201Tl SPECT in 1/1. These data suggest that 123I-IMT SPECT is a valuable addition to low grade glioma diagnostic and stress the need for a prospective study. PMID- 9743960 TI - Cerebellar mutism syndrome. AB - Since 1980, a growing number of pediatric patients with mutism following posterior fossa surgery have been recognized. This syndrome typically affects children and in rare cases young adults who become mute one or two days after tumor operation but do not show disturbances of consciousness or language comprehension. The disorder persists for 1 to 4 months. The pathogenesis is still unknown. Of 21 children who underwent surgery for large posterior fossa tumors between 1991 and 1995, 6 developed cerebellar mutism. Histologically the tumors were classified as astrocytoma WHO grade I, astrocytoma WHO grade II and ependymoma WHO grade III in one case and medulloblastoma WHO grade IV in three cases. Besides the clinical course, intraoperative findings and CT or MRI data are evaluated and discussed considering possible etiological hypotheses. Our own experience and also literature reviews suggest that the lesion of the cerebellar hemispheres might be the most important one of multiple factors causing cerebellar mutism. Generally the syndrome is transient. The diagnosis should not delay adjuvant therapy in patients with a malignancy. PMID- 9743962 TI - [Bacteremic episodes in pediatric oncologic patients, especially caused by the Streptococcus viridans group]. AB - BACKGROUND: The management of infectious complications plays a major role in the care for pediatric cancer patients. The majority of infections in the neutropenic patient present as fever of unknown origin without recovering a pathogen from the blood stream. The careful evaluation of bacteremic episodes is essential, since knowledge of the bacterial spectrum expected is crucial for the successful anti infective treatment. PATIENTS AND METHODS: From 1985 to 1995 all bacteremic episodes in pediatric oncology patients at the University Children's Hospital Freiburg were retrospectively analyzed with respect to the pathogens encountered, the antibiotic susceptibility profile and the underlying conditions. RESULTS: Overall, 113 bacteremic episodes were encountered in pediatric oncology patients, 68 of them in patients with hematological malignancies, and 45 in patients with solid tumors. In both patient groups, gram-positive bacteria were predominant with 72% and 58%, respectively. In patients with hematological malignancies, viridans streptococci were the most frequently isolated pathogens (35%) with a relevant morbidity (29% of patients developed a severe sepsis syndrome and/or ARDS), but were found only in 9% of patients with solid tumors. In 28% of patients with leukemia or lymphoma, gram-negative rods were cultured, in 6% Pseudomonas spec., in 4% Acinetobacter spec., and in 18% enterobacteria. In patients with solid tumors, in 38% gram-negative rods were isolated, 7% Pseudomonas spec., 16% Acinetobacter spec., 16% enterobacteria. In 3 patients, fungemia was observed. The antibiotic susceptibility profile was quite favorable in both, gram-positive and -negative bacteria: none of the gram-positive isolates was resistant to vancomycin, none of the Staphylococcus aureus isolates was resistant to oxacillin. All gram-negative bacteria were fully susceptible to ceftazidime, imipenem and ciprofloxacin. CONCLUSION: Gram-positive bacteria account for 2/3 of all bacteremic episodes in pediatric oncology patients. Streptococcus viridans was the most important pathogen in hematological malignancies accounting for a significant, pathogen-specific morbidity. PMID- 9743961 TI - [High dose chemotherapy with thiotepa, carboplatin, VP16 and autologous stem cell transplantation in treatment of malignant brain tumors with poor prognosis. Results of a mono-center pilot study]. AB - More than half of the children and adolescents with malignant brain tumors will relapse following initial therapy. Irrespective of the therapeutic modalities the prognosis of patients with recurrent or metastatic brain tumors is still poor. New strategies such as high dose chemotherapy (HDCT) with autologous blood stem cell transplantation (ABSCT) offer the possibility to improve the longterm prognosis of these patients. Following conventional chemotherapy with carboplatin/etoposide and after achieving complete or partial remission (CR or PR) 10 patients aged from 3.2 to 25.5 years (median, 10.3 years) with refractory or recurrent malignant brain tumors (anaplastic astrocytoma/glioblastoma, n = 2; medulloblastoma/PNET, n = 6; ependymoma, n = 1; plexus carcinoma, n = 1) received in a pilot study one course of HDCT with ABSCT. The consolidation regimen consisted of thiotepa (400-600mg/m2/d, i.v. 6 h, d-9), carboplatin and etoposide (500mg/m2/d, CVI 24h, d-8 to d-5, respectively) and was followed by the retransfusion of autologous blood stem cells on day 0. Before starting HDCT 6 patients showed CR and 4 patients had PR or stable disease (SD). Following the HDCT 3 of the 4 patients with residual tumor had CR or PR. 6 patients have remained in continuous CR or SD 8 to 41 months (median 17.2 months) after the HDCT. 2 patients relapsed 8.5 and 9.5 months after HDCT and died from progressive disease. Two patients died therapy-related from systemic aspergillosis and were not evaluable for response. Hematological recovery with an absolute neutrophile count of > 0.5 x 10(9)/l and a platelet count of > 30 x 10(9)/l was reached on days +11 (median; range, +9 to +14) and +16 (median; range, +6 to +47), respectively. The main nonhematological toxic effects were infections, severe mucositis, and hyperbilirubinemia. Although the long-term efficacy of HDCT with ABSCT is still not evaluable and the toxicity of this regimen is high, a multicenter phase II trial seems to be justified in view of the poor prognosis of recurrent or refractory brain tumors in children and adolescents. PMID- 9743964 TI - Lipid formulations of amphotericin B: clinical perspectives for the management of invasive fungal infections in children with cancer. AB - During the past four decades, amphotericin B deoxycholate has been the cornerstone of systemic chemotherapy for life-threatening fungal infections. Despite a broad spectrum of antifungal activity, its utility is greatly hampered by renal toxicity and limited clinical efficacy, in particular in patients with profound and persistent neutropenia. The novel lipid formulations of amphotericin B have distinct physicochemical properties resulting in different distribution patterns. Nevertheless, they all share a considerable reduction of nephrotoxicity, which allows for the delivery of higher daily dosages of amphotericin B. Preliminary efficacy data indicate that these compounds are overall at least as effective as amphotericin B deoxycholate. However, information for children is limited, and comparative studies for their use as first line agents are only in their beginnings. In this article, we review the clinical pharmacokinetics, safety, and efficacy of the lipid formulations of amphotericin B with special emphasis on pediatric data, and we seek to provide a rational framework for the determination of their current role in patients with cancer and proven or suspected invasive fungal infections. PMID- 9743963 TI - Vancomycin-resistant-enterococci--colonization of 24 patients on a pediatric oncology unit. AB - BACKGROUND: Colonization with multidrug-resistant vancomycin-resistant enterococci (VRE) could become a serious problem, since there is no proven therapy in case of an infection or in case of transfer of glycopeptid-resistance to other organisms. PATIENTS: Description of 24 from 48 pediatric oncology patients with VRE-colonization. METHODS: Stool samples were taken from all patients of our pediatric oncology unit from March 1996 until June 1997. Barrier isolation was introduced in May 1996, a prudent use of glycopeptid antibiotica in July 1996. RESULTS: 193 stool sample examinations demonstrated that 24 (50%) of the 48 patients were colonized with VRE. 11 (46%) of these 24 patients were VRE carriers at the time of their first examination; 9 (37%) patients acquired VRE during their therapy and 4 (17%) patients had come from other hospitals and already were VRE-positive when they entered our unit. In March 1997, one year after the outbreak only four patients still were VRE-positive, in June 1997 all of them were VRE-negative. The average time of colonization was 12.5 weeks. 17 (70%) of the 24 colonized patients had received glycopeptide antibiotics, 16 of them within two months before the appearance of VRE in their stool. Five colonized patients died, four of them because of their oncological illness, one because of a sepsis without proof of VRE in his blood. In the end none of our patients suffered from a VRE-infection, and besides that, the transfer of glycopeptid-resistance to other organisms was not observed. CONCLUSION: With barrier isolation and a very restrictive use of glycopeptid-antibiotica, colonization can be decreased and even stopped. Inspite of the high number of colonized patients no VRE-infectious disease occurred. PMID- 9743965 TI - [Clinical aspects and epidemiology of hepatitis C in immunosuppressed children with mostly oncologic diseases]. AB - Between July and October 1996 hepatitis C virus infection was diagnosed in 21 children who underwent immunosuppressive therapy mainly for malignant diseases. We report on the clinical signs and symptoms, diagnostic procedures and the clinical course of the disease in these patients. Epidemiological, diagnostic and clinical aspects of the outbreak are discussed. Analysis of all available data led to the conclusion that these infections were of nosocomial origin. This requires consequences in the hygienic regimen. In addition to the routinely used antibody-test the HCV-PCR should be the diagnostic method of first choice concerning the HCV-diagnostics in immunocompromised patients. PMID- 9743966 TI - Impaired renal function and tumor lysis syndrome in pediatric patients with non Hodgkin's lymphoma and B-ALL. Observations from the BFM-trials. AB - OBJECTIVE: Tumor lysis syndrome and renal failure remain important complications early in the course of therapy for pediatric Non-Hodgkin's lymphoma (NHL), frequently leading to therapeutic alterations. In the presented series, children with NHL and tumor lysis syndrome are retrospectively analysed regarding clinical features of acute renal failure and its implications on therapy. PATIENTS AND METHODS: From 4/1990 to 10/1997, 1192 patients diagnosed of any form of NHL have been registered in the NHL-BFM trials. 63 of these patients were reported to have suffered from impaired renal function and/or tumor lysis syndrome before or during initial treatment. Clinical data of these patients were analysed regarding diagnosis, stage, tumor mass, therapy and complications. RESULTS: 62 of 63 patients with impaired renal function and/or tumor lysis syndrome were diagnosed of Burkitt's lymphoma or B-ALL; 58 (92%) of these patients had advanced stages of disease and high LDH-levels (> 500 U/l). 43 of 63 patients had already signs of impaired renal function at admission. Hyperuricemia was the commonest cause of impaired renal function. Renal infiltration or enlargement was observed in 27 of 63 patients. 6 patients were diagnosed of urinary tract obstruction. 25 patients required hemodialysis. Despite improved renal function before administration of MTX, 21 of 63 patients suffered from protracted MTX-elimination during the first course of therapy. 7 of 63 patients (11%) with tumor lysis syndrome died of sepsis after the first course of therapy, another two patients died within 48 hours of therapy due to electrolyte imbalances. CONCLUSIONS: In pediatric NHL patients, Burkitt's lymphoma and B-ALL appear to be the commonest cause of metabolic complications early in chemotherapy. Patients with advanced stages and large tumor mass are at high risk for renal failure. Impaired renal function predisposes patients to further complications and toxic death. Prophylactic use of urate-oxidase in all patients with advanced stage NHL might limit the incidence of tumor lysis syndrome. Prospective studies on renal function prior to and during therapy are required in order to develop a clinical profile reliably detecting patients at risk for developing renal failure and subsequent complications. PMID- 9743967 TI - Coagulation and fibrinolysis in children with acute lymphoblastic leukaemia treated according to the COALL-05-92-protocol. AB - The etiology of thrombo-embolic events after therapy with asparaginase (ASP) is not fully understood. To investigate if cytotoxic drugs given in combination with asparaginase (ASP) have an additional effect on the coagulation system, a detailed analysis of coagulation factors was performed. Therefore, we investigated two combinations of the COALL-05-92-protocol, [cylophosphamide]/methotrexate/ASP ([CYC]/MTX/ASP) and high dose arabinoside/ASP (HIDAC/ASP). In 33 children with acute lymphoblastic leukaemia the following parameters were determined: plasminogen-activator-inhibitor-1, plasminogen, antiplasmin, protein C, antithrombin, modified antithrombin, prothrombin fragments 1 + 2 and thrombinantithrombin-complex. All children had an indwelling catheter. The most distinctive result of this investigation is that plasminogen shows a deeper and longer lasting decrease in the [CYC]/MTX/ASP-combination (median 65% NHP) compared to the HIDAC/ASP-combination (median 105% NHP) (p = 0.01). The other parameters showed the characteristic changes after ASP-therapy. None of our patients developed any clinical signs of thrombosis, even though two patients showed four altered coagulation parameters on the same day. This shows, that the coexistence of indwelling catheters plus four decreased coagulation parameters does not necessarily imply the development of thrombosis. We conclude that the parameters measured in this study do not sufficiently predict the development of thrombosis. PMID- 9743968 TI - Knowing what the others know: a study on interprofessional communication between nurses and medical doctors. AB - Communication and cooperation between experts from different specialties is a necessary element of professional competence. We have set up a research program to investigate cognitive aspects of interprofessional and interdisciplinary communication. We assume that successful communication essentially relies on communicants' ability to take the perspective of the other, i.e., assessing the background knowledge, expertise, plans, and so on, of one's co-communicators. The present paper reports on an interview study with medical doctors and nurses from three paediatric oncology units. The research interview was especially designed to allow for a detailed assessment of the professionals' ability to take their work-partners' perspective. Two areas will be examined: The ability to assess the work demands resulting from parental questions, and the use of differences in professional perspectives for prognosis. With respect to parental questions, doctors' and nurses' assessments of their respective work partner's task demands turned out to be only partly correct. Furthermore, the results suggest that doctors do not effectively benefit from nurses' experiential knowledge about patients when venturing prognoses. Educational measures designed to improve interprofessional communication could help to overcome these deficits. PMID- 9743969 TI - [Nursing care planning--that also! Presentation at the 50th Scientific Session of the Society of Pediatric Oncology and Hematology within the scope of the of the KOK AND PSAPOH general session]. AB - The relevance of structured planning and documentation in caring for chronically ill patients within the framework of a holistic nursing approach is emphasized. The author's claims are based on his experience in such kind of structured approach during two years of practicing as a paediatric nurse in the paediatric hospital of Hamburg/Eppendorf where such planning and documentation are encouraged and accompanied by supervisory support. The focus lies on the realisation of planning and documentation in nursing on the ward, on the requirements in terms of co-operation, and the consequences in terms of positive effects on a holistic approach towards patient care. PMID- 9743970 TI - [No motivation, no time, no money--does oncologic graduate and continuing education still have a future? Presentation at the 50th Scientific Session of the Society of Pediatric Oncology and Hematology within the scope of the KOK and PSAPOH general session]. AB - For many years the nursing profession has been considered a dead-end street without any prospects of career advancement or continued qualification. This situation has thoroughly changed. Nursing as an academic specialty has been established in 24 colleges. Furthermore, on-the-job specialty training has become available. In some regions of Germany the installation of specialty training courses has, however, been hampered by political opposition. The Northrhine Westphalian situation is described to illustrate the reasons for such an attitude. At the same time many nurses lack motivation to continue on-the-job training. Benefits (such as advancement opportunities) for those who complete training might serve to increase the motivation and participation of nurses. Specialty training is essential in ensuring the quality of care particularly in high pressure fields such as oncology and also extends the nurses' personal range of experience, reasons enough to enforce the legal realisation of specialty training. PMID- 9743971 TI - [Rehabilitation of families of children and adolescents with cancer--long-term psychosocial outcome]. AB - 24 families of a child with cancer were questioned for a 3rd time, 8.4 years after initial diagnosis and 6.5 years after family-oriented rehabilitation, before and after which they had been interviewed twice. Subject to this follow-up was the current family situation and the stability of the remarkably positive effect the rehabilitation had shown in the 1st course of the study. Also we hoped to be able to identify problem-families. Generally the family situation seems to have returned to normal, the standardised tests applied showed no significant difference to the general public. Nonetheless some families could be identified as having severe coping difficulties with 2 to 4 members displaying psychosocial problems of a very high degree. Both groups of families profited in the same degree from the rehabilitation; this effect shows great stability for the families coping good, whereas those with coping difficulties worsen considerably, almost always beyond the degree displayed before the rehabilitation. To identify these families quickly and offer them support it seems necessary to establish an out-patient psychosocial follow-up. PMID- 9743972 TI - [Congenital subglottic laryngeal stenosis in 2 brothers with chondrodysplasia syndrome (Keutel-Gabriel syndrome)]. AB - BACKGROUND: Keutel-Gabriel syndrome (chondrodysplasia) is a rare autosomal recessive disease. The patients have characteristic malformations such as midfacial hypoplasia, brachytelephalangia, and hearing loss as leading symptoms. PATIENTS: We report about two brothers with clinical and radiological features of Keutel-Gabriel syndrome. Congenital subglottic laryngeal stenosis was also present in both. In the younger brother an emergency tracheotomy had to be performed. In a staged procedure the stenosis was successfully treated with laryngotracheoplasty according to Cotton. CONCLUSIONS: This is the first description of a congenital subglottic laryngeal stenosis with Keutel-Gabriel syndrome. To avoid long-term tracheotomy, a tracheoplasty with autologous cartilage should be performed. PMID- 9743973 TI - [Dislocation of the cricoarytenoid joint: diagnosis and therapy]. AB - BACKGROUND: Laryngeal joint injury or cricoarytenoid dislocation is a relatively rare laryngologic finding, according to the international medical literature. It may occur as a result of external neck trauma or more frequently as a result of intubation. Chief symptoms are hoarseness, vocal fatigue, and loss of voice control. PATIENTS: Between 1993 and 1997 we diagnosed an arytenoid dislocation in 2 female and 5 male patients, in every case the etiology was an intubation trauma. Videolaryngoscopic recording was the most useful aid in diagnosis. RESULTS: Six patients were treated with closed reduction surgery between 8 and 49 days after dislocations. Normal voice was restored in four patients, and in one patient as late as 49 days after the dislocation. One patient had an additional recurrent nerve paralysis. CONCLUSIONS: Our results suggest that a closed reduction of the arytenoid luxation can be successful even several weeks after the injury. PMID- 9743974 TI - [Long-term results after modified Rethi-plasty in childhood]. AB - BACKGROUND: Laminotomy as described by Rethi has proved useful in the treatment of laryngotracheal stenoses. Using this technique in children poses the question as to whether it interferes with the growth of the larynx and trachea. The aim of our study was to verify the influence of this operative procedure on the structure and function of the restored airway. PATIENTS: Fifteen children between the ages of 2 and 16 with severe laryngotracheal stenoses (Cotton Grade III and IV) underwent reconstruction with interposition of a piece of costal cartilage in the posterior cricoid plate. Indications for this procedure included the presence of severe circumferential subglottic stenosis in combination with stenosis of the posterior glottis (8 children) and complete atresia (4 children). RESULTS: In endoscopic follow-up examinations (2-12 years follow-up), complete epithelization with light stenosis in the subglottis was demonstrated in all cases. Airway ventilation tests showed a slight or moderate central obstruction in nearly all cases. No disturbance of growth of the larynx was noted. CONCLUSIONS: This technique is a safe and successful method for repairing severe subglottic stenoses in children. Although there are some structural and functional problems during the long term follow-up, this is not considered a significant disadvantage. PMID- 9743975 TI - [Development and clinical testing of a non-magnetic cochlear implant. Preliminary experimental studies and surgical concept. Results in the first 10 patients]. AB - BACKGROUND: To ensure good transmission of electromagnetic signals from the speech processor to the internal cochlear stimulator (ICS) in cochlear implants, exact alignment of the external transmitter coil over the receiver coil of the ICS is necessary. Usually this is achieved by two magnets: one implanted within the ICS, the other one integrated into the transmitter coil of the external headset. Although this principle works, there are some serious problems with the implanted magnet. The most serious of these is that it renders MRI studies potentially hazardous or impossible, or at least compromises image quality. METHOD AND RESULTS: We developed a method to eliminate the magnet. The method requires varying the implantation technique and using a special headset. The technique is easy to perform, and the headset is suitable for series production. Experience with our first patients revealed the coil alignment to be remarkably stable, even more than in patients with a cochlear implant equipped with a magnet. Results of the first ten patients are presented. DISCUSSION: Aside from eliminating the magnet, we attempted to optimize the acoustic properties of the headset. This resulted in design changes such as repositioning the microphone. Further improvements such as integration of telephone coils and special sockets to connect peripheral appliances can easily be implemented in our special headset. PMID- 9743976 TI - [Comparison of methods for reducing residual noise in suprathreshold early auditory evoked potential registration]. AB - BACKGROUND: The level of residual noise in auditory brainstem responses (ABR) depends not only on the number of averages but also on the amplitude of background noise and on the frequency of artifacts. This paper describes the influence of digital filtering and of different methods of artifact suppression on the residual noise of ABRs. METHOD: Amplitude of background noise was estimated for 1033 ABRs recorded under suprathreshold stimulation (70 and 90 dB nHL) in 251 subjects. In 45 ABR recordings in 15 subjects, all 4000 individual sweeps were stored for off-line simulation. The power spectrum of background noise was investigated using an FFT analyzer. RESULTS: A great variability of mean noise amplitude was found both between subjects and in the recordings for each subject. Depending on the slope of the analogue 100-Hz high-pass filter, mean RMS values of background noise of 4.2 microV (6 dB/Oct.) and 2.5 microV (12 dB/Oct.), respectively, were found. Digital high pass filtering before averaging was found to increase the signal-to-noise ratio (SNR) considerably. CONCLUSIONS: Results indicate that (i) effective suppression of low-frequency noise components can only be achieved by zero phase digital filtering and (ii) if clipping of noise amplitude to 25 microV is used, optimized artifact rejection as weighted averaging or adopted artifact rejection levels have only small effect on the SNR. PMID- 9743977 TI - [Nasal fractures--indications for open reposition]. AB - BACKGROUND: Many septoplasties and septorhinoplasties are indicated due to previous traumatic etiologies, and the typical treatment of these septal and/or bone fractures has been closed reduction. Review of the literature reveals a success rate of only between 30-82%. One of the reasons for these poor outcomes is the influence of tension and pressure stress vectors due to hidden cartilage fractures behind intact septal mucosa. This is the main indication for open reduction of nasal trauma. The present study analyses the results of our experience with the open reduction of nasal fractures. METHODS: In a six-year period between 1991 and 1996, a total of 34 open reductions were performed out of 155 nasal fractures. This series was analysed clinically and with the aid of computer tomograms. Postoperative follow-up involved a combination of clinical evaluation, nasal endoscopy, and photography. The indication for open reposition was based on analysis of force of impact and suspicion of cartilage damage. The goal of the procedures was to incise partial-thickness fractures to make them full-thickness, thereby relieving the inherent tension in the cartilage, and to mobilize impacted bone fragments with microosteotomies so they could be properly reduced. At the same time, any previously existing anatomical abnormalities with functional impact were also corrected. RESULTS: The rate of patient satisfaction based on postoperative cosmesis and function was 88%. The group in which open reduction was most commonly indicated was the frontal impact cohort (n = 18.53% of open reductions). This was followed by the frontolateral impact cohort (n = 8.23%). In only one (1/155) open reduction was it not possible to find a cartilage fracture on exploration. Revision surgery was required in 2 cases due to nasal obstruction from synechiae between the septum and the anterior aspect of the inferior turbinate. Reoperation was necessary in a third patient due to airway obstruction from septal deviation. In a final case the patient was dissatisfied with the postoperative nasal appearance but declined revision surgery. Analysis of computer tomograms gave no additional information, though three-dimensional CT can aid in preoperative assessment of fragment position in those cases with severe edema and hematoma. CONCLUSION: Due to the high rate of subjective and objective success in postoperative nasal function and appearance, we suggest that consideration be given to widening the current indications for open reduction of nasal fractures. It is important to maintain a high degree of suspicion and explore the septum for subclinical fractures and be aware of the pattern of damage that can be anticipated based on classification of impact as described herein. The importance of precise clinical evaluation by inspection, palpation, and endoscopy cannot be overemphasized, and may not be replaced by radiographic imaging. PMID- 9743978 TI - [Atypical localization of a small cell carcinoma in the paranasal sinus area- case report]. AB - BACKGROUND: Malignant neoplasms of the paranasal sinuses are estimated at 3 to 5% of all head and neck malignant neoplasms. More than 50% of the cases are classified as squamous cell or anaplastic undifferentiated carcinomas. Extremely rare are small cell carcinomas localized in the paranasal sinuses. METHOD AND PATIENT: A 60-year-old male patient was seen in February 1996 in our ENT Department with unspecific pain on the left maxillary sinus and alveolar ridge. Anterior rhinoscopy revealed an extended tumor on the left nasal fossa; histopathological examination showed a small cell carcinoma. No other primary tumors or metastases were detected in extended staging. Due to the extended paranasal tumor as well as the histopathological findings, the patient was given induction chemotherapy followed by radiation therapy. RESULTS: To date (4/97), we achieved partial remission without any clinical complaints. CONCLUSIONS: The therapeutical result is comparable to other therapeutical regimens. PMID- 9743979 TI - [Complications with permanent damage in endonasal paranasal sinus operations- more frequent in experienced surgeons?]. AB - BACKGROUND: The rate of serious complications in endonasal sinus surgery has not gone down although optical aids are widely used nowadays. Are serious complications caused more often by unexperienced or experienced surgeons using a microscope and/or endoscope? METHODS: We defined serious complications as follows: death, persistent neurological deficits or permanent loss of vision, and injury to the internal carotid artery. Two different studies were made: the first consecutive 300 interventions of 6 sinus surgeons were evaluated. Sixteen malpractice cases were analysed regarding the experience of the surgeon. RESULTS: In 9 out of 16 malpractice cases serious complications were attributable to experienced surgeons, five to moderately experienced surgeons, and only two to an inexperienced surgeon (although he had extensive experience in external sinus surgery). There were 6 deaths, 6 neurologic defects, 2 visual disorders, and 2 injuries to the internal carotid artery without any sequelae. In 9 cases the serious complications were related to injury of the internal carotid artery, in five cases to perforation of the skull base. Twice the orbital wall was penetrated. In 1800 procedures performed by 6 surgeons, no serious complications were encountered. There were only lesions of the periorbit (n = 33) or CSF leaks (n = 8) without any permanent damage to the patient. CONCLUSIONS: Even an experienced surgeon must always keep in mind that serious complications can occur in sinus surgery. One must constantly be alert to the possibility of anatomical variants or specific pathologic findings. PMID- 9743980 TI - [Complications of endonasal paranasal sinus surgery--diagnostic and therapeutic consequences]. AB - BACKGROUND: Functional endoscopic sinus surgery has been proven the therapeutic method of choice in surgical therapy of chronic sinusitis. On the other hand, endonasal sinus surgery may cause severe complications even when performed by a skilled surgeon. This is easily explained by the close vicinity of many functionally important structures to the operative site. CASE REPORTS: Three histories are reported that involve possible complications even in apparently simple cases. Diagnostic and therapeutic consequences are discussed. In a case previously diagnosed histologically as chronic unspecific sinusitis, an endonasal biopsy resulted in endocranial bleeding requiring neurosurgical intervention. Midline granuloma was found to be the correct diagnosis. Another patient was seen with a normal X-ray of the sinuses and solitary polypoid structure in his left nose. Polypectomy was planned and a CT scan was performed, which demonstrated a meningocele. Transfacial surgery was then performed to remove the meningocele. Another patient presented with a traumatic impression of the frontal sinus, and open reposition by transfacial surgery of the frontal and ethmoid sinus was planned. When CT scans revealed an uncovered optic nerve in the sphenoid sinus of the fractured side, we abandoned ethmodectomy and performed reposition of the frontal sinus as the only surgical procedure. RESULTS AND CONCLUSIONS: In this paper, we show typical complications of endonasal sinus surgery and strategies for avoiding them. If any complication occur, prompt treatment is required. Three groups of complications can be defined: perforation of frontobasal dura resulting in cerebrospinal fluid (CSF) fistula, severe bleeding, and orbital or optic nerve injury. When the surgeon discovers an intraoperative complication, possible consequences must be considered immediately to minimize side effects for the patient. A CSF fistula should be closed in the same procedure, and transfacial surgery may be necessary. Hemorrhage resulting from an ethmoidal artery may require frontoorbital surgery and ligation of this vessel. If retrobulbar hemorrhage caused by retraction of an ethmoid artery occurs, immediate intervention is necessary. Usually a transfacial approach, resection of the medial orbital wall and retrobulbar decompression are performed. In some cases lateral canthotomy may be the best way to drain haematoma and decompress the optic nerve. Subsequently, orbital revision and ligation of the retracted artery must be performed. Any delay can result in persistent visual loss. We conclude that the extranasal frontoorbital approach should be part of the residency training program in ENT departments. Any surgeon performing endonasal sinus surgery must be trained in transfacial emergency procedures, which should be part of anatomic preparations in teaching courses, thus avoiding severe damage in case of intraoperative complication. PMID- 9743981 TI - [Ameloblastoma of the maxillary sinus: an electron microscopy study]. AB - BACKGROUND: The morphological demonstration and classification of rare tumor types occurring in the region of the head and neck may involve a number of problems. PATIENT AND METHODS: In the following the exceptional case of a plexiform ameloblastoma located in the main nasal cavity is presented. The morphological peculiarities of this locally destructive tumor are revealed by light and electron microscopy. RESULTS: The light microscopic investigations show one-layered epithelial units which are interlinked in a plexiform manner and extend as far as up to the surface epithelium. The electron microscopic investigations show solid and loosened areas. The organelles are well developed, their number varied; rough endoplasmic reticulum is scarce. Glycogen incorporations occur regularly. The tumor cells are connected with a number of desmosomes. The cell surface is characterized by microvilli. CONCLUSIONS: The combination of different methods of investigation is sometimes justified for diagnostic purposes. Especially electron microscopic studies may serve to demonstrate decisive structural peculiarities to classify a certain tumor type. Complete surgical removal is recommended. Postoperative radiation is generally advisable. PMID- 9743982 TI - [Nasal splint--space holder for after-care in endoscopic paranasal sinus operations]. AB - BACKGROUND: Scars and synechiae in the middle meatus of the nose are dreaded late complications of endoscopic sinus surgery. The placement of a stent into the ethmoidal sinus for ventilation and drainage during and after the wound healing in the nose and the sinuses seems to be advantageous. METHOD: A nasal splint (Nasensplint, Primed, Halberstadt) made of silicone, consisting of an oval shaped plate and a lateral groove with an attached strap is presented. A part of the groove should be placed in the middle meatus of the nose. The plate also acts as a stent after septal surgery. Up to now the nasal splint was placed in 32 patients after endoscopic sinus surgery and were left in place for 5-14 days. RESULTS: All patients tolerated the stents. No side effects or complications were observed. In the postoperative controls (2-12 weeks) no synechiae were observed. CONCLUSION: The nasal splint presented here seems to be a useful solution for problems in postoperative wound healing after endoscopic sinus surgery. It can help to avoid fibrin bridges between corresponding wound surfaces. PMID- 9743983 TI - [Guidelines/algorithms of the German Society of Otorhinolaryngology, Head and Neck Surgery]. PMID- 9743984 TI - [Interesting case no. 14. Penetrating injury of the left paranasal sinus system caused by a metal nail]. PMID- 9743985 TI - Do efflux pumps hold the key to better antifungal treatment? PMID- 9743986 TI - Targeted immunosuppressant on trial for rheumatoid arthritis. PMID- 9743987 TI - Blocking apoptotic damage after a heart attack. PMID- 9743988 TI - Noise-induced hearing loss: the future is hear. Cochlear pharmacology and noise trauma: prevention and progress-a joint symposium organized by The Novartis Foundation and the European Commission Concerted Action Against Noise. The Novartis Foundation London, UK, 1-2 May 1998. PMID- 9743989 TI - Pre-eclampsia: a disorder of placental mitochondria? AB - Pre-eclampsia is a common, pregnancy-induced, multisystem disease leading to severe complications in the mother and foetus. The aetiology of pre-eclampsia remains a mystery, but a growing body of evidence suggests that a mitochondrial defect might cause the impairement of differentiation and invasion of the trophoblast that leads to this disorder. This hypothesis is the topic of ongoing studies that, if confirmed, would be highly relevant to preventative strategies for this disease. PMID- 9743991 TI - Wiskott-Aldrich syndrome: a gene, a multifunctional protein and the beginnings of an explanation. AB - Patients with Wiskott-Aldrich syndrome show various defects in the normal function of platelets and lymphocytes. The recent identification of the gene responsible for this syndrome has led to a surge of studies aimed at solving the puzzle posed by the varied phenotype observed in this disease. It is now known that WASP, the protein product of this gene, can interact with a large number of other proteins known to be involved in the regulation of signal transduction and cytoskeletal organization. Thus, WASP appears to integrate these two basic and fundamental cellular mechanisms. Several groups are now focusing on understanding the function of WASP in detail, and translating this new knowledge into improved therapies. PMID- 9743990 TI - Prospects for gene therapy for cystic fibrosis. AB - Despite advances in conventional treatments for cystic fibrosis (CF), the disease is still associated with significant morbidity and mortality. The cloning of the cystic fibrosis transmembrane conductance regulator (CFTR) gene and the understanding of the functions of the CFTR protein have led to the development of novel treatment strategies, including gene therapy. Here, we review the underlying molecular defect in CF cells, and the progress in gene-transfer studies from in vitro work through to clinical trials. We discuss the problems encountered, the end-points used to assess efficacy, and the likely future directions of the field. PMID- 9743992 TI - Novel methods to monitor antigen-specific cytotoxic T-cell responses in cancer immunotherapy. AB - Monitoring cytotoxic T lymphocyte (CTL) responses to tumor antigens that have been defined at the molecular level has become essential to assess novel approaches to the specific immunotherapy of cancer. Nevertheless, because of the low affinity of the interactions between T-cell receptors and their ligands, there are no straightforward, well-standardized methods to meet this need. In this review, we describe several novel methods to track antigen-specific CTL responses. PMID- 9743993 TI - The genetic basis of tuberous sclerosis. AB - Tuberous sclerosis is a relatively common inherited disease that causes multiple benign tumours in different organs, frequently leading to skin rashes, seizures and mental handicap. The disease can be caused by mutations in either of two genes, TSC2, identified in 1993, and TSC1, only recently identified. Here we review the current state of knowledge of the molecular genetics of tuberous sclerosis and the spectrum of mutations seen in and the implications of recent findings for patients. Although both genes appear to function as tumour suppressors, the function of their protein products is not understood. A speculative model of how these proteins might function is briefly described. PMID- 9743994 TI - [Research for the genes of epilepsy]. PMID- 9743995 TI - [Anterior interhemispheric approach to the diseases involving the third ventricle and/or the suprasellar region]. PMID- 9743996 TI - [Management of elderly patients with incidentally discovered unruptured aneurysms]. AB - Long-term natural history of unruptured cerebral aneurysms is not found frequently. Hence, the indications for surgery on unruptured asymptomatic cerebral aneurysms are still unclear. The benefit of treatment ultimately depends on the relative risk of subsequent aneurysm rupture in untreated patients versus the risk involved in surgery. Clinical features of fourteen elderly patients aged over 70 years with incidentally discovered unruptured aneurysms were analyzed. Two were male and 12 were female, with ages ranging from 70 to 82 years (mean: 74.5 years). Aneurysms were located in the anterior circulation in 13 patients and in the posterior circulation in 1 patient. One patient had multiple aneurysms, that is, bilateral middle cerebral aneurysms. The size of all these aneurysms was less than 10 mm. The indication for surgery was determined case by case. General information about the natural history of incidentally discovered aneurysms was given to the patients and their relatives. Informed consent was based on the fact that subarachnoid hemorrhage was associated with a poor prognosis, while excellent operative results were common in patients with unruptured aneurysms. Five patients agreed to surgical treatment. Four of them, their ages being 70, 70, 72 and 72 years old, with aneurysms located on the middle cerebral arteries, underwent neck clipping of their aneurysms with no operative morbidity or mortality. However, the remaining one patient was not recommended for surgery in spite of her consent to having it, because of her high age (82 years) and the location of her aneurysm (intracavernous internal carotid artery). Consequently, 10 patients who didn't receive surgery were followed-up at periods ranging from 3 months to 7 years. Two patients developed rupture of the aneurysms, either proven (one patient) or presumed (one patient). The former patient made an uneventful recovery after surgery, but the latter died. None of the remaining eight patients have experienced rupture of the aneurysms. It is our clinical impression, however, that they harbor an unruptured aneurysm with at least mild trepidation. With the rapid aging of the population, withholding aneurysm surgery merely because a patient is elderly may not necessarily be the most appropriate decision. Our conclusions are as follows: (1) Elderly patients in their early seventies are apt to agree to having surgical treatment for their unruptured aneurysms. (2) The cases reported herein show that asymptomatic middle cerebral artery aneurysms were able to be clipped very safely. (3) Most patients have experienced a decrease in quality of life from knowing they are living with an unruptured aneurysm. PMID- 9743997 TI - [Clinical experience of autologous blood transfusion and fibrin glue in neurosurgery]. AB - Clinical experience of autologous blood preservation on 83 patients and transfusion on 43 patients were reported. When drawing blood, 14 patients whose hemoglobin level was below 13 g/dl (body weight < 70 kg) or 14 g/dl (body weight > 70 kg) received subcutaneous recombinant human erythropoietin injection (s.c.) to facilitate erythropoiesis, according to the internal standard protocol. Hemoglobin levels of all the patients recovered to more than 10 g/dl by the time they were admitted to the hospital, which value would not interfere with general neurosurgical procedures. The injection of erythropoietin did not cause any side effects. Autologous blood transfusion was performed in 43 patients but, in 3 patients, additional homologous transfusion was required because of excessive bleeding. Except in cases with meningioma, postoperative hemoglobin values were identical with preoperative values, indicating that autologous blood transfusion was enough to replace intraoperative blood loss. Autologous fibrin glue was applied in 74 patients. In 70 cases including 55 with skull base surgery, the glue was applied to ensure dural closure. The incidence of cerebrospinal fluid leakage was 16.4% (5 patients) in skull base surgery. This incidence was identical to or less than that in previous reports. The glue was also effective in transposing and fixing offending vessels in 4 cases which received microvascular decompression. As a conclusion, procedures for autologous blood preservation and transfusion were safely performed in neurosurgical cases. Review of the literature was also presented to discuss the advantages and problems to be solved in the future. PMID- 9743998 TI - [A study of acute subdural hematoma developing into hematoma with capsule formation]. AB - There are some cases in which conservatively treated acute subdural hematoma (ASDH) does not disappear naturally and progresses to chronic subdural hematoma like hematoma (CSDH) (hematoma with capsule formation). The objective of the present study was to identify factors which can be used to predict this unfavorable course during the early phase after the onset of the lesion. During the past 13 years, 10 of 96 cases of mild, conservatively treated ASDH (excluding suckling infants) progressed to CSDH, and those 10 patients showed the following background characteristics. There were 7 males and 3 females, and the mean age was 63.1 years. Five of the patients had a history of alcohol consumption, and one case each had a history of cerebral infarction, cerebral hemorrhage and a VP shunt. Acute-phase computerized tomography (CT) at the time of ASDH showed, in all 10 cases, an expansive-type lesion with a low density area in the hematoma, with expansion of the hematoma into the interhemispheric fissure. The hematoma was observed to undergo transient natural shrinkage in the acute phase. The period for progression to CSDH was indicated to be a mean of 20.5 days after the onset of the lesion, and its cure was possible with trepanation. In consideration of these results, it was surmised that ASDH patients with the following characteristics have a high risk of progression to CSDH during the subacute and chronic phases when conservative therapy is administered during the acute phase of the lesion: (1) old age, (2) a history indicative of brain atrophy, (3) an expansive-type image of ASDH on acute-phase CT, and (4) acute-phase CT indicative of cerebrospinal fluid mixing in the hematoma. PMID- 9743999 TI - [Venous system playing a key role in transpetrosal approach]. AB - A case with a large vertebrobasilar junction aneurysm developed a venous infarction in the temporal lobe after an operation using the transpetrosal approach. Although little of the literature has been concerned with venous complications after the transpetrosal approach, the case prompted us to study the venous system as playing a key role in the transpetrosal approach. Analyzing 30 carotid and 15 vertebral angiograms of 15 patients who underwent preoperative cerebral angiography using digital subtraction angiography (DSA), we investigated the venous system near the junction of the superior petrosal sinus, the transverse sinus and the sigmoid sinus (STS junction) which may play a key role in the transpetrosal approach. Drainage pathways of the superficial middle cerebral vein (SMCV) were classified into four types; sphenoparietal, sphenobasal, sphenopetrosal and undeveloped. In the sphenopetrosal type (4/30: 13%), the drainage of SMCV passes back along the floor of the middle fossa to drain into the transverse sinus. The lateral temporal vein (LTV) and the temporobasal vein (TBV) drain into the transverse sinus. The LTV emptied into the transverse sinus either directly (20/30: 67%) or indirectly through the tentorial sinus (10/30: 33%). The entry of the LTV into the transverse sinus (venous point) was usually located in the lateral third of the transverse sinus (14/20: 70%), but sometimes in the middle third (6/20: 30%). The TBV, observed in 8/30 (27%), also often emptied into the tentorial sinus to drain into the transverse sinus. Atresia of a unilateral transverse sinus and a large LTV emptying into the distal sigmoid sinus was observed in 2/15 cases. The venous system near the STS junction may be interrupted by the incision of the tentorium and the middle fossa dura mater and by the retraction of the sigmoid sinus. Since the transpetrosal approach may cause venous complication by compromising the venous system near the STS junction, it is necessary to evaluate of the venous system preoperatively using DSA and to set up a surgical strategy preserving the venous system. PMID- 9744000 TI - [Stereotaxy during intravenous anesthesia with propofol]. AB - A series of 30 patients, who underwent stereotactic surgery for movement disorder under intravenous propofol anesthesia between March, 1995 and December, 1997, was retrospectively reviewed. In 28 patients with Parkinson's disease including seven juvenile cases of parkinsonism, the postoperative motor and ADL scores on the Unified Parkinson's Disease Rating Scale significantly improved. In the other two patients, one of whom had severe posttraumatic tremor and the other had cerebral palsy, the stereotactic surgery produced considerable alleviation of their symptoms. We evaluated and discussed the usefulness of intravenous propofol anesthesia in stereotaxy. Except for one patient who had an allergic reaction against propofol, none of the patients complained of intraoperative pain postoperatively. Wake-up tests were performed to record neural noise levels in 26 cases. This recording was performed under propofol anesthesia in two cases with advanced Parkinson's disease and one with cerebral palsy. In these patients, neural noise levels were recorded and were useful for identifying the target. Although the tremor disappeared under propofol anesthesia in 17 patients presenting with moderate or severe tremor, it was presented again after discontinuation of propofol. Wake-up test, therefore, made a good evaluation of Vim thalamotomy for tremor. In juvenile parkinsonian patients, three presented with dopa-induced dyskinesia (DID) during propofol infusion. In two of them, the DID emerged immediately after posteroventral pallidotomy and continued 4 or 10 hours after stereotaxy. These findings suggest that propofol possibly has an anti parkinsonian effect. Intravenous propofol anesthesia is a useful method to use with stereotactic surgery for movement disorders. PMID- 9744001 TI - [A case of a lumbar spinal synovial cyst located on the midline]. AB - We report a case of lumbar spinal synovial cyst located on the midline. A 72-year old man was admitted to our hospital with the chief complaint of low back pain radiating to the left buttock and posterior thigh. An MR image revealed an extradural cystic lesion adjacent to the dorsal side of the dural sac at the L4-5 level. The cyst was remote from the facet joints and existed on the midline just in front of the L4 lamina. A CT scan showed a concave deformity of the ventral aspect of the L4 lamina because of compression by the cyst. The patient underwent L4 laminectomy and total removal of the cyst. The cyst was in contact with the anterior surface of the ligamentum flavum and it had no connection with the facet joint. In the histological examination, the cyst was multilobular and lined with synovial epithelium. Therefore the cyst was diagnosed as a synovial cyst. After the operation, the pain radiating to the buttock and thigh completely disappeared. Intraspinal synovial cysts are usually located in the lower lumbar spine and most of them are adjacent to the facet joint. The cysts that are located on the midline are very rare. We review previous reports and discuss clinical and pathological features of spinal synovial cysts. PMID- 9744002 TI - [Multilocular chronic intracerebral hematoma due to metastatic brain tumor: a case report]. AB - Chronic intracerebral hematoma is a fairly rare clinical entity. We report a case of a 76-year-old man who presented with left hemiparesis on admission. CT scan revealed a hematoma in the right basal ganglia. Under observation, the hyperdense lesion on CT scan changed to isodense 2 weeks after admission and then changed to low density 4 weeks after the first scan. These findings suggested hypertensive intracerebral hemorrhage but, 3 months after admission, hemiparesis grew worse and the appearance of the lesion changed from low density to isodensity again. When the patient presented disturbance of consciousness, CT scan revealed a new hemorrhage from the capsule of the hematoma and the new hematoma markedly expanded 2 weeks later. The contents of the hematoma was old uncoagulated hematoma resembling chronic subdural hematoma and drainage was carried out under local anesthesia. The patient died because of respiratory failure and autopsy was performed. Histopathologically cancer cells were shown in the hematoma cavity and the capsule of the hematoma was composed of two layers: a collagenous layer on the inside and a granulation layer with neovascular system on the outside. As the neovascular system existed on the outside of the capsule, the rebleeding occurred outside the capsule and the second hematoma appeared beside the initial one. We suspected that the initial bleeding occurred in the tumor tissue and the rebleeding occurred in the granulation layer of the capsule. PMID- 9744003 TI - [Choroidal artery aneurysms of the posterior inferior cerebellar artery presented with fourth ventricular hemorrhage: report of 2 cases]. AB - Two cases involving a ruptured aneurysm in a choroidal branch of the posterior inferior cerebellar artery are reported here. Case 1: A 61-year-old woman was admitted after an episode of severe headache with persistent vomiting. A CT revealed an intraventricular hemorrhage within the fourth ventricle. An angiography showed an aneurysmal shadow in the choroidal artery branching from the telovelotonsillar segment of the distal posterior inferior cerebellar artery (PICA). The operation disclosed a fusiform aneurysm in the choroidal artery which was successfully trapped using Yasargil's mini-clips. The postoperative course was uneventful and the patient was discharged without any neurological deficit. Case 2: A 64-year-old woman became unresponsive after complaining of a severe headache. On admission, she was semicomatose with positive bilateral Babinski's sign. A CT scan showed that the fourth and third ventricles were packed and dilated by a massive hematoma. An angiography demonstrated an aneurysmal shadow in a branch from the PICA with an occlusion of the right vertebral artery. Furthermore, the left vertebral artery was also occluded and the basilar artery was fed by collateral circulation. The patient underwent an operation immediately. The fusiform aneurysm was resected after ligation. Her postoperative course was satisfactory. She was able to go home without neurological deficit. There has been only one article about "pure" choroidal artery aneurysm, reported by Uranishi, et. al in 1994. They suggested that the pathogenesis of this lesion could be due to hemodynamic stress. Our two cases also present the same characteristics, in the shape of the aneurysms as well as in the anomalous structures in the posterior circulation. Our results offer further evidence concerning the pathogenesis of that type of lesion. PMID- 9744004 TI - [Rapid growth of glioblastoma during therapy for multiple myeloma: case report]. AB - Rapid growth of a glioblastoma during therapy for multiple myeloma is reported. A 53-year-old man was admitted to our hospital with a right costal tumor, which was resected. The diagnosis was plasmocytoma. Urine protein electrophoresis showed a monoclonal peak in the region of gamma-globulin, and examination of the bone marrow revealed 17.8% of atypical plasma cells. Brain magnetic resonance (MR) imaging detected two small lesions, but these could not be identified as brain tumor. He received chemotherapy (melphalan 10 mg/day and predonin 30 mg/day for 4 days) and was discharged. Two weeks after discharge, he was readmitted because of left hemiparesis. T1-weighted MR imaging showed two large hypointense lesions in the right frontal lobe, with ring-like enhancement following Gd-DTPA infusion. 1H MR spectroscopy showed typical findings of tumor with increased choline and lactic acid peaks. 201Tl SPECT revealed high accumulation in both early and delayed images. Right carotid angiography showed a hypervascular tumor with venous filling and mass effect. The lesions were resected via right frontal craniotomy, followed by intraoperative radiation and placement of an Ommaya reservoir. Histological examination showed the tumors were glioblastoma. The brain between the tumors also showed the typical appearance of glioblastoma, suggesting that the lesions were continuous. Postoperatively, the patient's left hemiparesis disappeared. He received local irradiation and chemotherapy and was then discharged. Coexistence of glioblastoma and multiple myeloma is rare. The cause may be genetic abnormality, but immunodeficiency due to multiple myeloma, surgical damage, or chemotherapy may have contributed to the rapid growth of the glioblastoma. PMID- 9744005 TI - [Meningioma associated with acute subdural hematoma: a case report]. AB - We present a case of meningioma associated with acute subdural hematoma. This 67 year-old male had a sudden onset of severe headache when he was on the train. He had a CT scan which revealed an acute subdural hematoma at the left parietal convex. Cerebral angiography disclosed a small focus (3 x 4 cm) of vascular stain under the left parietal bone supplied by the left middle meningeal artery. He was diagnosed as having a meningioma with surrounding acute subdural hematoma. The removal of this tumor was carried out without delay. It was fragile and the bleeding point was not detected. Pathological diagnosis was meningothelial meningioma. The literature showed meningioma associated with acute subdural hematoma is rare, but when it is discovered incidentally, surgical resection might be indicated. PMID- 9744006 TI - [Bone marrow infiltration in neuroblastoma]. AB - Bone marrow infiltration if documented at diagnosis in the majority of children with neuroblastoma. Despite its chemosensitivity it is difficult to eradicate. In addition, current methods to detect it following treatment are inadequate. The following two articles report improved results in evaluating the minimal residual disease in the bone marrow. The prognostic and therapeutic relevance will be presented and discussed. PMID- 9744007 TI - [The morphological assessment of bone marrow infiltration in neuroblastoma]. AB - Bone marrow biopsy is very important in diagnosis and follow-up of children affected by neuroblastoma (NB). Between June 1995 and May 1997 we studied 55 patients with NB stage 4. Specimens were obtained at the diagnosis (in 8 patients) and after chemotherapy (in 55 patients) in order to evaluate the effects of treatment on bone marrow disease. 88% of 343 biopsies were representative versus 99% of 639 aspirates. Of 8 stage 4 patients evaluated at diagnosis, 15/16 biopsies versus 9/15 aspirates were positive. Following chemotherapy, out of 298 evaluable sites examined, 111 biopsies versus 30 aspirates (37 versus 10%) were positive. Of 111 positive biopsies 53 showed a focal pattern (35 differentiated, 18 undifferentiated), while 51 showed a diffuse pattern (18 differentiated, 40 undifferentiated). Our results confirm previous literature data indicating a better efficacy of histology versus morphology in detecting residual bone marrow disease (especially in case of focal differentiated pattern). The recent introduction of a specific monoclonal antibody, called anti-GD2, has improved our capacity to detect minimal residual disease in patients' bone marrow. The inclusion of anti-GD2 immunohistochemistry in our evaluation will possibly increase our overall sensitivity to detect minimal residual disease and may provide information capable to direct the physician's decision into a more rational patient's treatment. PMID- 9744009 TI - Early and late postoperative complete heart block in pediatric patients submitted to open-heart surgery for congenital heart disease. AB - The incidence of complete heart block (CHB) following open-heart surgery for congenital heart disease is about 1%. Most of postoperative CHBs are the consequence of procedures involving the closure of ventricular septal defect; they usually occur immediately after surgery or early in the postoperative period; in few cases they also may occur many months or years after surgery. Early postoperative CHB can be transient or permanent. Permanent pacing is generally not recommended in the former. On the contrary, if CHB persists after at least two weeks of temporary pacing, permanent pacing is needed because the block is usually due to His bundle damage or to trifascicular damage and this is associated with excessive bradycardia and risk of asystole. Late postoperative CHB can be due to the recurrence of previous transient early postoperative CHB or to the progression of postoperative His-Purkinje conduction troubles suggesting trifascicular damage. Permanent pacing is obviously needed in case of documented late postoperative CHB. The prophylactic use of permanent pacing in patients at risk of late postoperative CHB is still a controversial point. Electrophysiologic studies should be performed in such patients. The occurrence of second degree AV block within or below the bundle of His during atrial pacing at rate lower than 200/min can be considered a good marker of impending CHB. In this case prophylactic permanent pacing should be recommended, especially in patients with coexisting problems of troublesome or malignant tachyarrhythmias who have to be treated with antiarrhythmic drug therapy that may favour the progression to CHB. PMID- 9744010 TI - [Microscopic hematuria in children]. AB - Hematuria is a common finding on a urinalysis, with a prevalence rate between 1% and 2%. The execution of screening of hematuria in children is controversial. Once hematuria has been identified, it is useful to identify sources of bleeding, as either glomerular or non-glomerular. If microscopic hematuria is confirmed, investigations would include: hypercalciuria screen, blood examinations (full blood count, renal function tests, complement and autoantibody screen), renal tract ultrasound, urinalysis of family members, audiogram, if family history of deafness is present, or family members present a positive dipstick. If all these tests are negative and microscopic hematuria persists, then a renal biopsy is advocated. PMID- 9744008 TI - [The identification of minimal residual disease in the bone marrow and peripheral blood in neuroblastoma. The prognostic and therapeutic implications]. AB - A highly sensitive and specific methodology to detect neuroblastoma cells in the bone marrow and peripheral blood of children with neuroblastoma is of critical importance for proper staging and treatment of these patients. In addition, patients with bone marrow infiltration at diagnosis need to undergo regular investigation to measure the effectiveness of chemotherapy (so called "in vivo" purging). Finally, the evaluation of autologous stem cells taken from bone marrow or peripheral blood is necessary to rule out or minimise the possibility of reinfusing tumor cells to the patient following myeloablative therapy. The authors provide a "state of the art" data on this complicated issue and give their preliminary results of their own experience, mainly concerning the immunocytological methods. PMID- 9744011 TI - [Normal levels of collagen-type-I telopeptide in the first 90 days of life]. AB - Serum levels of carboxyterminal telopeptide of type I collagen (ICTP), a marker of matrix degradation, were measured by RIA test, on 184 samples of healthy newborns and children aging from 1 (cord blood) to 90 days of life. We found ICTP values about tenfold higher than the adults', with highly significant variations (P < 0.001) in the whole period studied. During the first three months of life serum levels of the bone marker show a progressive increase from 0 to 7 days, they remain unchanged until the 30th day and then decrease until the 45th day, maintaining similar values from the 45th to the 90th day of life. The authors think that the pattern of ICTP in the first week of life is under the influence of the adapting phenomena following delivery, in which catabolic processes are predominant, while in the second period ICTP modifications are related to growing processes and then to bone turnover. PMID- 9744012 TI - [The importance of using the cerebral function monitor (CFM) in the neurological prognosis of neonates in intensive care]. AB - Cerebral function monitor (CFM), unlike traditional EEG, allows a long-term evaluation of electric brain activity, without interfering with the nursing of the newborn in the intensive care unit. Our aim was to evaluate the prognostic value of CFM for neurological outcome. We studied 102 newborns (gestational age 34.5 +/- 4.36 weeks; weight 1980 +/- 720 grams) by Multitrace CFM (Lectromed) 5 hours daily in the first week following admission. The patients also underwent cerebral echography, EEG and neurological follow-up to the 24th month. CFM was found to correlate well with the EEG recorded 3 months later. The persistence for at least one week of an I.C. tracing or the normalization of initial tracing have a good prognostic value (positive predictive value 95.23%), a persistently pathologic registration has a negative prognostic value (negative predictive value 85.18%), that even increases if cerebral echographic alterations are demonstrated (98.57%). The association of CFM and ultrasound abnormalities determines a relative risk for neurological motor impairment of 69.14, whereas CFM alone gives a relative risk of 6.4. PMID- 9744013 TI - [Glucose as an analgesic in neonatology. A blind randomized controlled study]. AB - The aim of this study is the evaluation of the effect of different oral glucose solutions on the pain response after heelstick in newborns. Four groups of healthy newborns (gestational age 38-41 weeks, weight over 2500 g) were randomly allocated to receive 2 ml of water (19 cases) or glucose 5% solution (17 cases) or glucose 33% solution (15 cases) or nothing (control group, 14 cases) immediately before heelstick procedure. The cardiac rate of each newborn was registered before, during and 3 minutes after the procedure. Glucose 33% solution reduces the pain response (p < 0.01). PMID- 9744014 TI - [Recurrent asthmatic bronchitis in the first years of life: a 3-year follow-up]. AB - The natural history of asthma is not well known; its origin may go back to just after the birth. The aims of this study are: 1) studying the evolution of the symptomatology of a group of children suffering from recurrent asthmatic bronchitis of a moderate-severe level in the first two years of life; 2) inquiring some risk factors conditioning the evolution of it. One hundred children, aged from 16.6 to 53.4 months, suffering from moderate to severe recurrent asthmatic bronchitis, have been checked at the follow-up at the Broncopneumologic and Allergic Pediatric Unit of the SS Annunziata Hospital (Naples). Among the 100 children checked, 25 have been dropped at the follow-up; the difference between the sample checked and the one dropped at the follow-up is not statistically relevant. Four are variables examined: age, allergic medical history, number of asthma episodes, breast feeding, association or not of atopic dermatitis. The evolution of the symptomatology has not been propitious, as, at the end of three years of analysis, 73.3% of the 5-year-old patients had still persistent asthma. The risk factors of asthma recurrence are: parents smoking habits with a relative risk of 18 times more in patients with disadvantageous evolution (R.R. = 18) and at a less measure the overcrowding of the bedroom (R.R. = 1.5). The age at the onset in of the symptomatology, the positive familiar medical history for allergic illness, the breast feeding and the atopic dermatitis have not resulted statistically different between the group with advantageous evolution and the one with disadvantageous outcome. Passive smoking is a very disadvantageous prognostic factor; it may contribute to the disadvantageous evolution of the moderate to severe asthmatic bronchitis in the early childhood towards the recurrent or chronic bronchial asthma of the late childhood. PMID- 9744015 TI - [The beta-hemolytic Streptococcus group A carrier state]. AB - In this study we describe the group A streptococcal upper respiratory tract carrier state, following a clinical method based on two main elements: the presence or the absence of symptoms and the result of throat swab. Through this method we are able to decide how to face each individual clinical case. PMID- 9744016 TI - [The cell-mediated response after measles vaccination]. AB - Natural measles virus infection is recognized for causing prolonged abnormalities in immune responses, that contribute to the severe and complicated evolution of the disease. Immunization with live measles virus vaccine could be considered a mild form of the measles infection. Results of investigation of in vitro immune response after measles immunization with live attenuate vaccine have been conflicting. In this work we studied cellular immune parameters in children aged between 3 and 12 years. T cells, CD4+ and CD8+ subsets were analyzed by cytometry. CD3+ cells were significantly reduced compared to controls (p < 0.01) whereas CD4+/CD8+ ratio was normal. The in vitro proliferative response to polyclonal mitogen was significantly reduced (p < 0.01). This study confirms the presence of a mainly functional immunosuppression of cellular response in a cohort of children belonging to a developed area. These findings improve the understanding of the mechanism of immune response to virus measles and provide suggestions for the development of a better approach to immunization, taking account for the strain, vaccine titre, age and environmental conditions of the target population. PMID- 9744017 TI - [Children at social risk and the utilization of health services]. AB - To investigate the association between social risk factors and access to emergency health services, 333 children, with a medium age of 39.1 months, have been recruited. 31.2% of these children had at least one social risk factor. After 12 months through a phone call we evidentiated that children with social risk showed an access to emergency services about double (relative risk factor more than double) in respect to children with no social risk factors. PMID- 9744018 TI - [The abuse of anabolic substances by an adolescent going in for sports]. AB - The authors describe the case of an elder adolescent who showed an imposing symptomatology due to the abuse of anabolic drugs. They urge the pediatricians to warn their patients about the use of such drugs. PMID- 9744019 TI - [The VACTERL association: a report of a clinical case with hepatic cystic lymphangiectasis]. AB - VACTERL association includes three or more of the following six anomalies: V (vertebral anomalies), A (anal atresia), C (cardiac abnormalities), TE (tracheo esophageal fistula and/or esophageal atresia), R (reno-urinary anomalies) and L (limb defects). VACTERL cases are classified as "associated" when other than the typical six defects are present in the same infant, or "isolated" when they are not. We report a case of VACTERL association "associated" that presents an hepatic cystic lymphangiectasia that was never described before in literature. We also consider the most important factors involved in the aetiology of the typical anomalies. PMID- 9744020 TI - [Infantile acute hemorrhagic edema (IAHE). A report of a new clinical case]. AB - The authors describe a new case of infantile acute haemorrhagic oedema in a six month-old child. Even being a vasculitis the authors do not retain to assimilate it to the Schonlein-Henoch syndrome. PMID- 9744021 TI - [Neonatal hypocalcemia]. PMID- 9744022 TI - Pain in the very preterm baby: 'suffer little children?'. AB - Increases in medical expertise and technological advances have enabled the survival of very preterm babies who form a new and growing population. Comparisons between the foetus, full-term baby and the very preterm baby indicate that by the time the foetus is of 23 weeks gestation, many of the abilities, for example, sensing touch, hearing, seeing, moving and even learning may be common to all three. Thus, the very preterm infant who has been described as a unique organism, is not passive, but is a sentient being who is unlikely to survive without the medical and technical support of the Neonatal Intensive Care Unit (NICU) where he/she is exposed to frequent and regular medical procedures. Many of these procedures would be, for any normal, fully developed human being at best uncomfortable and at worst painful. Reviews within the past 10 years have shown that the neurochemical, anatomic and functional systems of newborns are developed enough to perceive pain. More importantly, rat pup studies have indicated that not only may the very preterm baby experience pain but it may experience it more intensely than the more mature infant. Moreover, there may be serious consequences of repeated painful medical intervention. Alleviation of pain and/or distress in very preterm infants is, therefore, an important issue. PMID- 9744023 TI - Reaction and movement time variability in ADD/H: effect of tactile experience. AB - A clinical group of children with attention deficit disorders with hyperactivity (ADD/H) was compared to a control group, each divided into high and low variability (HV and LV) on reaction time (RT) and movement time (MT) measures. The effects of tactile-somatosensory experience on performance was also investigated. Analyses of the HV and LV groups based on RT variability found improvement on MT in both HV groups (ADD/H and control) following a tactile somatosensory task. Both HV groups based on MT variability also showed improvement in MT following the TPT administration. No LV group (based on either RT or MT) showed improvement in MT or RT. The results in this study indicated that HV (either RT or MT) predicted improved speed and accuracy following a tactile somatosensory task. Several neuroanatomical models for the study of response variability and the role of tactile somatosensory training programmes in paediatric rehabilitation are discussed. PMID- 9744024 TI - Late outcome after severe traumatic brain injury in children and adolescents. AB - OBJECTIVES: Eighteen surviving adolescents with severe traumatic brain injury were re-examined a mean period of 7.1 years after their trauma in order to determine their life situation, motor, cognitive functions and pattern of handicap. METHODS: A structured interview, the EB test of motor function, Ravens's progressive matrices, Peabody's neuropsychological test, SPIQ and the WHO classification of handicap were used. RESULTS: The group had a mean WHO Classification of Handicap score of 1.61 (SD 1.60) revealing mild handicap, and performed as a group significantly subnormal (p < 0.0001) in gross motor, fine motor, sensibility and perception sub-tests. The EB test revealed a mean value of 2.23 (SD 0.89) corresponding to mild disability. The mean non-verbal IQ score of 93.1 (SD 13.9) and the verbal score of 93.4 (SD 14.8) were within normal limits. CONCLUSIONS: Only 28% of the group of surviving adolescent TBI victims functioned within normal limits. The most crucial disabling component was poor social integration, which was clearly demonstrated in the WHO score. PMID- 9744025 TI - Long-term ventilator-dependent children: a vocal profile analysis. AB - PURPOSE: Ventilator dependent children need to be able to communicate effectively to be able to re-integrate into society. This study was performed to see if the Vocal Profile Analysis can be used to assess the voice quality produced by speaking aids in ventilated children, and therefore help to direct speech therapy input. METHODS: This study attempted to evaluate vocal function using a perceptual rating scale in four ventilator-dependent children fitted with the Passy-Muir Tracheostomy Speaking Valve. The children were at different stages of speech development and had either intact or abnormal cognitive function. The evaluation was performed independently by three trained speech therapists blinded to the child's underlying diagnosis. RESULTS: Although speaking aids enhanced verbal communication skills, most of the subjects still displayed marked abnormalities of speech. There was significantly greater agreement among the trained observers when assessing children who were cognitively intact at the time of the study. The older child with intact cognitive function achieved near normal speech potential with the speaking valve. CONCLUSIONS: The Vocal Profile Analysis is a useful tool for assessing vocal characteristics in ventilated children with intact cognitive function. Near normal speech can be attained with speaking aids, provided a certain stage of speech development has already been attained. PMID- 9744026 TI - Implementation of a compensatory memory system in a school age child with severe memory impairment. AB - Impairments in memory and new learning secondary to neurological illness or injury pose significant problems for school age children. This paper describes the development and implementation of compensatory memory aids for an adolescent girl who demonstrated significant memory impairment following irradiation for an intracranial tumour. Strategies for integrating the system with the student's academic programme and school-related activities, and involvement of school personnel are emphasized, as is a theoretically and behaviourally based training programme. PMID- 9744027 TI - Using differential reinforcement to treat functional hypophonia in a paediatric rehabilitation patient. AB - Behaviour analysis studies have demonstrated the use of operant procedures for treating symptoms of psychiatric disorders. These symptoms are usually caused or maintained by a variety of variables including specific organic mechanisms, yet most can be modified by environmental social contingencies. In this case study, data is presented on a 16 year-old male admitted to a rehabilitation hospital following the onset of polymyositis. This patient also presented with functional hypophonia and met the DSM-IV diagnostic criterion for a conversion disorder. In conjunction with the medical team and a speech pathologist, a behavioural programme was developed and implemented to shape and differentially reinforce increasingly complex vocalizations. Results were evaluated using a moving treatment, multiple baseline across responses design. Differential reinforcement in the form of written and verbal feedback was effective in shaping normal speech. The protocol was applied comprehensively across staff and settings. Results are discussed in terms of basic behavioural research on 'learned non use'. PMID- 9744028 TI - The diagnostic yield of the nerve-muscle skin biopsy in paediatric neurology practice. The Montreal Children's Hospital Neuromuscular Group. AB - OBJECTIVE: To determine the diagnostic yield of the nerve-muscle-skin (NMS) biopsy in paediatric neurology practice. STUDY DESIGN: A consecutive series of 98 paediatric NMS biopsies done 1989-1994 retrospectively reviewed in the context of pre-biopsy clinical and laboratory parameters. Bivariate associations based on chi-square test. Unconfounded associations between pre-biopsy variables and positive diagnostic yield (PDY) assessed by multiple logistic regression. RESULTS: Fifty seven out of 98 patients central (global delay, seizures, abnormal CNS imaging) process; 41/98 patients peripheral (motor delay, weakness) process, electromyography-nerve conduction studies (EMG-NCS) 87/98 cases; abnormal 43/87. Positive diagnostic yield (PDY) in 42/98 (43%) biopsies. Statistically significant bivariate associations between PDY and pre-biopsy; age, presenting symptom, developmental delay, weakness, reflexes, CPK, lactate, EMG-NCS and process. Unconfounded associations demonstrated with PDY and age, reflexes and process. The presence of a peripheral process or an abnormal EMG-NCS strongly predictive of PDY: 34/41 (83%) peripheral process cases had PDY, 32/40 (80%) abnormal EMG-NCS cases had PDY, and 29/31 (93.5%) peripheral process and abnormal EMG-NCS cases had PDY. Abnormal EMG-NCS with central process improved PDY to 3/9 (33%) from 4/37 (11%) for normal EMG-NCS. CONCLUSION: NMS biopsy is a valuable diagnostic tool, particularly in the context of a suspected peripheral process or a central processes with an abnormal EMG-NCS. PMID- 9744029 TI - Multidisciplinary rehabilitation management of depression in the carbon monoxide injured patient. AB - Carbon monoxide (CO) is one of the common inhaled intoxicants used in attempting suicide. This case report describes the multidisciplinary rehabilitation management of suicidal CO poisoning in a 13-year-old initially comatose male patient. Despite the prevalence of self-inflicted CO poisoning, this is the first detailed case report emphasizing the acute and delayed neuropsychiatric sequelae of CO poisoning in the premorbidly depressed patient. In addition, the difficulty of assessing psychiatric illness in an acutely neurologically impaired patient is discussed. PMID- 9744031 TI - [Evaluation of indirect immunofluorescence as a supplementary test for the diagnosis of HIV-1 infection]. AB - In order to be used as an alternative or complementary test to confirm HIV-1 infection, the efficiency of indirect immunofluorescence assay (IFA) was compared with Western blot (WB) in 362 samples from persons with high and low risk behaviour. A panel of sera with 220 WB positive, 122 WB negative and 20 WB indeterminate sera were tested by an "in house" IFA. The sensitivity of IFA was found to be 98.63% and the specificity 98.36%. Therefore, IFA appeared to be an efficient alternative method to WB, since the cost of testing by IFA is less than 10% of WB testing. We observed a direct relationship between WB protein reactivity and IFA results. In 15 samples with coincident indeterminate results for WB and IFA, antibody reactivity to p24 and gp160 presented the highest frequency. On the other hand, antibodies to viral glycoproteins were always present in IFA weak positive samples, showing their high predictive value. PMID- 9744030 TI - [Detection using molecular biology techniques of Mycoplasma hominis and Ureaplasma urealyticum in urogenital samples]. AB - Mycoplasma hominis and Ureaplasma urealyticum are species closely related to urogenital diseases such as pyelonephritis, nongonococcal urethritis, urinary calculi, epididymitis, pelvic inflammation, infertility, abortions and post delivery fever. They can also cause pneumonia and meningitis in newborn infants. In this paper we used nucleic acid hybridization and polymerase chain reaction to analyze 22 samples from patients with different urogenital symptoms in order to detect mycoplasmas and ureaplasmas. We obtained 10 positive samples and 12 were negative. From positive samples we identified two with Mycoplasma hominis, two with Ureaplasma and six with both species. The results obtained by these molecular techniques were compared with reference methods and we found coincident results in 18 samples, while in four the results were discordant. These discordant findings were not statistically significant. PMID- 9744032 TI - [Obtaining antibodies and development of an immunoassay for the detection of Escherichia coli proteins in preparations of human recombinant gamma interferon]. AB - The present paper refers to the obtainment of polyspecific antisera directed against Escherichia coli host strain used to produce recombinant human gamma interferon (rec. hum. gamma IFN). The antisera were obtained by the cascade immunization method. The animals (n = 3) were initially immunized with an E. coli protein preparation (EcPp) of the host strain obtained from a blank run which assures the absence of the rec. hum. gamma IFN protein. Afterwards, consecutive immunizations were carried out with the less immunogenic proteins. To obtain those proteins, EcPp is passed through a column containing antibodies purified from previous inoculations coupled to a gel matrix. In this way, the proteins that have not been recognized by the immune system in that moment (e.g. do not have their corresponding antibodies coupled to the column) are separated an used to reimmunize the animals. The analyses of the antisera by Western blotting show a progressive recognition of the host strain proteins by the antisera with the progression of the cascade method. The recognition is evident through all the molecular weight range. Those antisera were used as quality control of the recombinant protein, by quantification of the host strain protein contaminants using a multiantigenic ELISA. The detection limit of that system was 3.125 ng/mL and the quantification limit 6.25 ng/mL. PMID- 9744033 TI - [Chitinase production by a strain of Streptomyces griseoruber isolated from the rhizosphere of sugar cane]. AB - Twenty nine mesophilic and 19 thermophilic actinomycetes from sugar cane phyllosphere, rhizosphere and root-free soil were isolated. Twelve mesophilic and 7 thermophilic strains produced clear zones of hydrolyses on colloidal chitin agar and hydrolysed a chitin colloidal suspension. The rhizosphere strains were significantly more chitinolytic (p < 0.05) than the root-free soil strains. Streptomyces griseoruber 202 produced chitinase in colloidal chitin liquid medium with an activity of 108 micrograms of colloidal chitin hydrolysed/mg protein/hour, at pH 4.6 and 40 degrees C. This strain also produced chitinase in a medium with insect exoskeleton powder. PMID- 9744034 TI - [Dermatophytosis in the greater Resistencia area, Chaco Province, Argentina]. AB - Epidemiological characteristics of dermatophytoses in the metropolitan area of Resistencia city are described. Hair fragments, skin, scalp or nail scrapings were collected from 3.507 persons with dermatological symptoms of probable fungal origin. The mycological studies were performed by three mycological laboratories, one from a university Institute and two from private laboratories. Direct microscopic examinations showed fungi in 39.66% of samples, while the recovering of dermatophytes from cultures was 40.66%. Microsporum canis was the most prevalent species among dermatophytes, isolated in 217 samples (41.73% of cultures) while Trichophyton rubrum, Epidermophyton floccosum, Microsporum gypseum and Trichophyton mentagrophytes were present in 38.46%, 6.35%, 5.58% and 4.81%, respectively. Microsporum and Trichophyton genus were clearly predominant over Epidermophyton. Variations in prevalences of fungal genus from different places of the world probably reflect the existence of particular environmental situations defined by biotic and abiotic factors that influence the settlement of endemic mycoses. PMID- 9744035 TI - [Agglutination of hen egg-yolk immunoglobulins (IgY) against Salmonella enterica, serovar enteritidis]. AB - Two groups of 6 laying hens were used to produce IgY. In the vaccinated group (V), hens were injected by intramuscular route with two doses of a Salmonella enterica serovar Enteritidis bacterin at 20-day interval. In the control group (T) hens remained unvaccinated. Four IgY extractions were performed on the egg production of both groups. The first two extractions were carried out using the yolks obtained from the eggs produced during the 4th and 5th post-vaccination week (extracts 1V and 1T) and the other two using the ones from the 6th, 7th and 8th week (2V and 2T). Starting from the extracts 1V and 1T other products were obtained by freezing-thawing (1V-A and 1T-A) and simple (1V-B and 1T-B) or double (1V-C and 1T-C) flow capillary dialysis concentration. All these products were compared using an ELISA test specific for the detection of chicken antibodies against flagellar antigens of S. Enteritidis. In this test, V extracts were positive whereas T extracts were negative. The extract 1V was more positive than the extract 2V. The extract 1V-C was the most positive and was therefore selected to be used as an antiserum in the agglutination tests. This extract contained 1.9 g/dl of total proteins, 0.028 g/dl of triglycerides and 0.012 g/dl of cholesterol and showed an electrophoretic pattern characteristic of IgY. The 1T-C extract was used as a negative control in the agglutination tests. Slide somatic and tube flagellar agglutination tests were simultaneously carried out using both IgY extracts and a standard rabbit anti-Salmonella (IgG) sera. Overall 367 strains from the Enterobacteriaceae family were tested together with two other strains belonging to the Vibrionaceae family. The 1V-C extract specifically agglutinated S. Enteritidis strains in the same way as the rabbit sera. This extract also agglutinated other Salmonella strains antigenically related to S. Enteritidis. Salmonella which did not share somatic or flagellar antigens with S. Enteritidis, other different species of the Enterobacteriaceae family and the two strains of the Vibrionaceae family were all negative. None of the strains tested was agglutinated by the 1T-C extract. This paper show that it is possible to use specific IgY to identify S. enterica serovars. The more extended use of IgY for diagnostic purposes may be a convenient way to complement the current use of mammal polyclonal antibodies. PMID- 9744036 TI - [Isolation of Helicobacter pylori from dental plaque]. AB - It has been suggested that oral dissemination might be the major transmission vehicle for Helicobacter pylori, and that dental plaque might act as its reservoir. The presence of H. pylori was investigated in 62 odontological male and female patients (average age: 35 years old). Samples were taken from supragingival plaque, placed in 0.3 ml of thioglycolate broth, cultured within 12 h in Mueller-Hinton agar with the addition of 5-7% of sheep blood and antibiotic supplement, and incubated at 37 degrees C in microaerophilia for 5-7 days. Typical colonies were identified by gram, urease, oxidase and catalase. H. pylori was detected in a 15 year-old patient suffering from gastric acidity (1.61% positivity index). The medium used facilitated recovery of the agent from a sample abundant in germs. H. pylori was not recovered from the same patient 12 months later, suggesting that there might have been a transitory passage by gastric reflux or that the bacterium was acquired from an exogenous source. PMID- 9744038 TI - [Rapid differentiation and presumptive identificaiton of yeasts using Candida CHROM-agar medium]. AB - In order to evaluate faster and cheaper methods for isolation and identification of clinically important yeasts, we used CHROM-agar Candida (CAC), a chromogenic medium, for differentiation and presumptive identification. A total of 546 yeast strains were studied. Strains were previously identified by conventional methods. The colour and texture of Candida albicans, C. tropicalis, C. krusei and Trichosporon beigelii were always constant and particularly distinctive for a reliable identification. C. glabrata, Saccharomyces cerevisiae, C. parapsilopsis colonies also showed constant colour. Because of its contents of cloramphenicol, CAC is also suitable for isolation and observation of mixed yeast species from clinical samples. CAC is a useful medium for reliable differentiation and presumptive identification of clinically important yeasts, particularly form immunocompromised patients. PMID- 9744037 TI - [Sewage sludges: application of a technique for recovering Enterovirus]. AB - The aim of this work was to develop a technique to recover viruses from sewage sludges and to set a microbiological reliable index to control the efficiency of waste-water disinfection methods. Twelve samples were collected at San Felipe treatment plant, where the waste-water from Tucuman, Argentina, is received. To free virus from solids, sludges were shaken during 20 h at 4 degrees C and supernatants were obtained by centrifugation. Penicillin, streptomycin, neomycin and amphotericin B were added to avoid the development of contaminating flora. Samples were inoculated in Vero and HeLa cells and Enterovirus (Echo 7, Echo 11 and Echo 21) were isolated from five samples. PMID- 9744039 TI - [Ultrasound morphological evaluation of adnexa tumors]. PMID- 9744040 TI - A new sonomorphologic scoring system (Mainz Score) for the assessment of ovarian tumors using transvaginal ultrasonography. Part I: A comparison between the scoring-system and the assessment by an experienced sonographer. AB - OBJECTIVE: The problem of an accurate sonographic assessment of ovarian tumor status has not yet been solved. To what extent can the preoperative assessment of adnexal tumors be improved on the basis of a maximum number of sonographic tumor parameters included in the newly developed sonomorphologic Mainz Score? MATERIALS AND METHODS: In a prospective study 314 premenopausal patients with adnexal tumor underwent a transvaginal sonographic examination performed by an experienced sonographer. In parallel to the sonographic examination a new score including 10 different sonographic parameters was used to predict adnexal tumor status: 1. Total tumor structure, 2. tumor border, 3. wall thickness, 4. inner echos in cystic component, 5. septa, 6. shape of echo complex or of the completely solid tumor, 7. echogenicity of the echo complex or of the completely solid tumor, 8. acuostic phenomena behind tumor, 9. ascites, 10. detection of liver metastases/peritoneal carcinosis Depending on the respective degree of expression, the individual characteristics were rated on a scale from 0 to 2. The total score obtained following addition of the points recorded for each parameters served to confirm the validity of the sonographic tumor status assessment. The first sonographer assessed the tumor status based on his experience and in the knowledge of all clinical parameters. The second sonographer evaluated the tumor status based on the score. All preoperative ultrasonographic findings were compared with the postoperative histologic analysis. RESULTS: A maximum number of 20 points may be obtained using the Mainz Score. Tumors with a total score of below 9 were rated as benign and those with a score of above 9 as malignant. This resulted in a sensitivity of 96.4%, a specificity of 80.7%, a positive predictive value of 47.4%, and a negative predictive value of 99.6%. The predictive value of the scoring-system was diminished by the presence of 30 false-positive cases, which were identified as inflammatory conglomerate tumors, teratomas, endometrial cysts, cystadenomas and hemorrhagic cysts. The experienced sonographer assessed 233 cases as benign and 24 cases as malignant. The findings were confirmed by the histological examination in 252 of 257 cases. No conclusive prediction of tumor status could be made in 57 tumors. However, the application of the Mainz Score enabled an accurate prediction of the tumor status in 44 of the 57 cases. With the exception of septal thickness all assessment criteria of the score showed a statistically significant correlation between the assigned score and the histologic findings, (p < 0.05). CONCLUSIONS: The use of the Mainz Score enables even less experienced sonographers to assess the status of premenopausal adnexal tumors with a high degree of accuracy. The score provides the experienced sonographer with a refined and improved method for the prediction of tumor status, especially in the presence of not readily assessable findings. PMID- 9744041 TI - [Doppler ultrasound of uterine and fetoplacental circulation: placental laterality, normal values and reference curves]. AB - AIM: Doppler ultrasound assessment of blood flow velocity waveforms from the maternal and fetal sides of the placenta in normal pregnancies, taking into consideration placental location, in order to establish normal values and reference curves. METHOD: A cross sectional study of uneventful singleton pregnancies with gestational ages from 18 to 41 weeks and later on healthy newborns was performed by continuous-wave-Doppler ultrasound. Doppler flow signals from the uterine artery or the corresponding ascending branch and the umbilical arteries were analysed using the resistance index RI. The RI-values from the uterine circulation were classified according to the proximity of the placenta. Normal values and reference curves were established on the basis of centiles. RESULTS: After consideration of exclusion criteria, 757 pregnancies remained for evaluation. The resistance index reference curves of the uterine circulation were dependent on placental location, with proximal RI-values being lower, but with nearly constant course during the second half of pregnancy. In contrast, the umbilical artery RI-values were higher but showed a continuous decrease. CONCLUSION: In the second half of pregnancy, down stream flow resistance of the uterine circulation is very low and nearly constant but depends on placental location. The fetoplacental circulation shows high flow resistances which decrease with increasing gestational age. These reference curves are comparable with others where placental location is taken into consideration. PMID- 9744042 TI - [The supratrochlear artery as an indicator of cerebral hemodynamics in carotid occlusion]. AB - AIM: In internal carotid artery (ICA) occlusions a reversal flow in the ophthalmic artery (OA) has been considered both an important collateral as well as a sign of inadequate intracranial cross-flow. The aim of the study was to evaluate the effect of OA collateral on the cerebral haemodynamics using ultrasound and CCT. METHODS: In 540 patients with a total of 577 ICA occlusions the presence of an OA collateral was investigated with periorbital continuous wave Doppler sonography and the findings were correlated with the ipsilateral cerebrovascular reserve capacity (CVR), measured with the Transcranial Doppler (TCD) CO2 test. Moreover, in 128 of 577 ICA occlusions the presence of an OA collateral was compared with the type of infarction in CCT and the neurological deficit. RESULTS: In 577 ICA occlusions an OA collateral was found in 59.4% (n = 343), whereas there was no or anterograde OA flow in 40.6% (n = 234). A significant correlation could be demonstrated between the CO2 reactivity and an OA collateral, which was present in 51.3% ipsilateral to a normal CVR (n = 175), in 66.9% ipsilateral to a diminished CVR (n = 115) and in 82.8% ipsilateral to an exhausted CVR (n = 53). Patients with OA collateral had a 15.5% risk of developing an exhausted CO2 reactivity, while there was only a 4.7% risk in the remaining patients (p < 0.001). In the 23 patients with haemodynamic infarctions in CCT the highest percentage of an OA collateral (87%) was found; no correlation could be demonstrated in these cases between the degree of neurological impairment and OA collateral flow. CONCLUSION: Patients with ICA occlusions and an OA collateral are subject to a more than a threefold risk of suffering from an exhausted CO2 reactivity in comparison with those revealing no reversal OA flow. Nevertheless, in patients with ICA occlusions and an OA collateral, CO2 reactivity was found to be normal in 51%, implying that in individual cases ultrasonic investigation of the periorbital flow is in itself insufficient to estimate the haemodynamic stroke risk and therefore cannot replace the TCD CO2 testing. PMID- 9744043 TI - [Transparent 3-D ultrasound in fetal abnormalities]. AB - AIM: Evaluation of three-dimensional ultrasound (3D-ultrasound) for optimal diagnosis in fetal malformations. METHOD: The 3D-system used consists of a transmitter, position sensor and 3D-workstation. In 38 fetal malformations, verified using real-time two dimensional ultrasound, 101 volume scans were recorded to prove the following functional abilities of the system; (1) Sequential analysis of single images including volume estimation; (2) Simultaneous three plane display (longitudinal, transverse and horizontal planes); (3) Cubic sectional image; (4) Rotating or swinging volume block. RESULTS: In 85% of the cases the 3D-ultrasound clearly achieved a more reliable diagnosis in comparison to the two-dimensional images. Furthermore, the diagnostic information was considerably improved in 44% of the investigated malformations compared to the conventional two-dimensional method. The main advantages of the transparent 3D-technology are (1) Plastic global imaging of the pathological topography (fetal tumours, ureteral stenosis); (2) Exact measurement of irregular volumes (ventricle system, cerebellum); (3) Precise evaluation of fetal disproportions (triploidy, osteochondrodysplasia); (4) Detection of minor malformations. The disadvantages of the method is that all fetal movements during the scanning procedure affect adversely the three-dimensional display. CONCLUSION: The application of 3D-ultrasound in specialized prenatal units recommended. PMID- 9744044 TI - [Time requirements of medical and non-medical personnel for ultrasound studies]. AB - To assess the time needed for ultrasound examinations for physicians and assistants we carried out a survey among members of the German Association of Ultrasound in Medicine (DEGUM) in hospitals and practices. The physician's examination time for the investigation of the upper abdomen and kidneys was 12.3 min with additional time for preparation (2.9 min), for work up (2.3 min) and written documentation (3.6 min), together 21.3 min. The examination times in practices (18.9 min) are shorter than in ultrasound labs in hospitals. Assistants in ultrasound laboratories need for organisation, upkeep of equipment, to filing the images in archives etc. medium 15.2 min per patient. Assistants with exclusive employment in the ultrasound laboratory need 17.6 min (4-57 min), whereas assistants employed in the ultrasound laboratory and other departments need 12.7 min (1.6-31 min). In internal practices the desired time is 7.9 min (3.5-20 min). Because of the different organisation in the ultrasound laboratories in several hospitals and in practices a comparison of times needed by assistant personnel (or, in many cases, by the physicians who perform such work themselves) is very difficult. The examination time really needed by physicians and the time for assistant personnel are shorter than presumed earlier, but are longer than estimated in the recent discussion about money saving in medicine. PMID- 9744045 TI - [Standard ultrasound sections of the upper extremity--shoulder and elbow joint]. AB - AIM: The sonographic examination of the shoulder and elbow joint is demonstrated in standardized sectional planes providing a reproducible representation of anatomical structures. METHOD: In ultrasound examination of the shoulder joint the following standardized planes are employed transverse and longitudinal planes in the dorsal, superior/lateral and ventral region. In ultrasound examination of the elbow joint the following standardized planes are employed: a transverse and two longitudinal planes in the ventral region, a transverse and a longitudinal plane in the dorsal region. These standardized sectional planes, performed in a real-time-procedure using a 7.5 MHz linear transducer, as well as the images on the screen, correspond to the guidelines provided by the German Society of Ultrasonography in the Locomotor Apparatus (DEGUM). In cases without pathological findings two standardized planes have to be documented. Otherwise the pathological finding has to be documented in two standardized planes. RESULTS: The introduced examination technique in standardized sectional planes provides an excellent overview of the shoulder and elbow joint with a sufficient evaluation of the periarticular soft tissue and the bony surface. The main advantages in comparison to other diagnostic means (i.e. CT, MRI) lies in the possible dynamic examination which allows excellent imaging especially in rotator cuff lesions. CONCLUSION: Because of the excellent evaluation of the periarticular structures, ultrasound plays a major diagnostic role in shoulder joint diseases. Furthermore this technique provides important information about intra- and periarticular disorders of the elbow joint. The standardized procedures are a useful means of assuring and improving the quality of sonographic examinations of articular and periarticular structures. PMID- 9744047 TI - [Doppler ultrasound of the umbilical vein in fetal 3rd degree atrioventricular block]. AB - A 21 week fetus was diagnosed with complete heart block. From the first diagnosis until 38 weeks gestation, umbilical venous pulsations occurred during ventricular systole. The ventricular frequency was 49 bpm. The venous pulsations corresponded with the ventricular frequency. At 38 weeks venous pulsations also occurred during ventricular diastole. Umbilical pulsations during diastole were associated with a decrease of ventricular frequency to 28 bpm and fetal hydrops. The venous pulsations during ventricular systole seem to be characteristic of complete heart block. The pulsations during ventricular diastole reflect the fetal heart failure. Doppler ultrasound aids in the diagnosis and surveillance of the fetus with complete heart block. PMID- 9744046 TI - Cardiac metastasis of a malignant fibrous histiocytoma occupying the right ventricle and infiltrating the myocardium. AB - A 86-year-old woman presented with a cardiac tumor occupying the whole cavity of the right ventricle and causing right ventricular outflow obstruction. A percutaneous ultrasound-guided biopsy was performed and histologic examination revealed a metastasis of a malignant fibrous histiocytoma. Because of the extensive cardiac involvement and multiple pulmonary metastases, effective surgical therapy was not possible and the patient died soon after diagnosis. PMID- 9744048 TI - Uterine artery Doppler velocimetry as a screening test: where we are and where we go. PMID- 9744049 TI - Telemedicine and ultrasonography: making waves. PMID- 9744050 TI - Ultrasonographic assessment of the uterine cervix and interleukin-8 concentrations in cervical secretions predict intrauterine infection in patients with preterm labor and intact membranes. AB - OBJECTIVE: Interleukin-8 concentrations in cervical secretions have been related to microbial invasion of the amniotic cavity and histological chorioamnionitis. Since a short uterine cervix may be a risk factor for intrauterine infection, we set out to determine the interrelationship between cervical-secretion interleukin 8 concentration, cervical length measured by transvaginal sonography and intrauterine infection in women with preterm labor and intact membranes. DESIGN: The study group comprised 144 pregnant women admitted to hospital for preterm labor and intact membranes. At admission, interleukin-8 in cervical secretions was assayed. The uterine cervix was evaluated by transvaginal sonography and the cervical index (funnel length + 1)/cervical length) was measured. In all cases, amniotic fluid was obtained by amniocentesis immediately after cervical examination and was cultured for aerobic and anaerobic bacteria including Ureaplasma ureolyticum and Mycoplasma hominis. Placentas were analyzed at delivery for the presence of histological chorioamnionitis. RESULTS: Forty-three per cent (62/144) of pregnancies delivered preterm and 12.5% (18/144) of the amniotic fluid cultures were positive. Placentas were available from 54 pregnancies which delivered preterm and histological chorioamnionitis was found in 59.2% (32/54) of the cases. Interleukin-8 concentrations in cervical secretions were significantly higher in the presence of microbial invasion of the amniotic cavity (median 1191.5 ng/ml, range 812-5234 ng/ml vs. median 109 ng/ml, range 12-2231 ng/ml; p < or = 0.0001) and histological chorioamnionitis (median 982.5 ng/ml, range 430-5234 ng/ml vs. median 435 ng/ml, range 40-1750 ng/ml; p < or = 0.0001). Similarly, higher values for cervical index were obtained in the presence of a positive amniotic fluid culture (median 1.23, range 0.35-5.88 vs. median 0.29, range 0.024-4.85; p < or = 0.0001) or histological chorioamnionitis (median 1.18, range 0.043-5.88 vs. median 0.562, range 0.040-2.48; p = 0.011). Multiple logistic regression analysis indicates significant independent associations with a positive amniotic fluid culture and histological chorioamnionitis for the cervical interleukin-8 concentrations (amniotic fluid culture positive > or = 850, histological chorioamnionitis > or = 450) and for the cervical index (amniotic fluid culture positive < or = 0.58, histological chorioamnionitis < or = 0.56). CONCLUSIONS: Intrauterine infection is associated with increased interleukin-8 concentrations in cervical secretions and a short cervix. Their combined analysis may reduce the indications for invasive procedures and improve the selection of women in preterm labor who may benefit from antimicrobial treatment. PMID- 9744051 TI - Fetal liver volume measurement by three-dimensional ultrasonography: a preliminary study. AB - OBJECTIVE: To establish the application of three-dimensional ultrasonography in measuring fetal liver volume during the second half of normal pregnancy. DESIGN: A prospective cross-sectional study of normal fetal liver volume at 19-39 weeks of gestation (median 29 weeks). SUBJECTS: Thirty-four non-smoking women with a singleton pregnancy resulting in the delivery of a healthy infant with a birth weight between the 5th and 95th centiles according to the Kloosterman tables adjusted for maternal parity and fetal sex. METHODS: For fetal liver volume measurements, a simultaneous recording of a frontal section of the liver immediately anterior to the stomach and a sagittal section of the liver were obtained using a standard Combison 530 ultrasound machine with a 5-MHz annular array transducer for volume scanning. RESULTS: Technically acceptable fetal liver volume measurements were obtained in 25 of 34 participating women. Mean fetal liver volume data (P50) ranged between 8 ml at 20 weeks' gestation and 116 ml at 38 weeks' gestation. A statistically significant increase in normal fetal liver volume was found with advancing gestational age (p < 0.0001) and with increasing estimated fetal weight (p < 0.00001). CONCLUSIONS: Three-dimensional ultrasound allows measurement of fetal liver volume, and this demonstrated an approximately 14-fold increase during the second half of pregnancy. It is speculated that three dimensional fetal liver volume measurement may identify the fetus at risk of growth restriction. PMID- 9744052 TI - Second-trimester measurements of placental volume by three-dimensional ultrasound to predict small-for-gestational-age infants. AB - OBJECTIVE: The objective of this study was to investigate the value of second trimester three-dimensional sonographic placental volume measurements to predict infants who are below the 10th centile for birth weight. SUBJECTS AND METHODS: Placental volume measurements were performed using three-dimensional ultrasound in 382 women with normal singleton pregnancies at 16-23 weeks' gestation. Logistic multivariate regression consisting of variables considered to influence birth weight was used to predict infants with a birth weight below the 10th centile from the placental volume measurements. RESULTS: Prognostic influence could be shown for placental volume (p < or = 0.0001), gestational week at the time of measurement (p = 0.0002) and maternal weight at the time of registration (p = 0.0025). Values for specificity and sensitivity achieved by choosing an 'optimal' cut-off point of 0.16 for the estimation probability for a small-for gestational-age (SGA) infant were low at 82.5% and 52.5%, respectively. CONCLUSIONS: Three-dimensional sonographic measurement of the placental volume alone is not a satisfactory technique for predicting SGA infants. PMID- 9744053 TI - Antenatal sonographic diagnosis of club foot with particular attention to the implications and outcomes of isolated club foot. AB - OBJECTIVE: There are no studies to date on the implications and outcomes of antenatally detected isolated club foot. The purpose of this study was to perform a contemporary evaluation of club foot diagnosed in the antenatal period. DESIGN: We performed a retrospective analysis of all ultrasound examinations performed in 1989-96 in the Fetal Diagnosis and Treatment Unit of the University of Iowa Hospitals and Clinics (n = 23,863). SUBJECTS AND METHODS: All cases of club foot (n = 35) were evaluated for the presence of other detectable abnormalities and karyotype results if available. Postnatal follow-up was performed until over 1 year of age. RESULTS: We diagnosed unilateral (n = 18) and bilateral (n = 17) club foot from 17.4 to 37.0 weeks. Defects involving other systems were found in 28 of 35 cases. Of the seven cases considered to be isolated antenatally, three were diagnosed with additional malformations in the neonatal period. CONCLUSION: Most cases of antenatally diagnosed club foot were not isolated. Even when they were thought to be isolated on antenatal ultrasound, over half of them were later found to be associated with additional severe abnormalities that were detectable only in the neonatal period. PMID- 9744054 TI - Perinatal outcome and prognostic factors in prenatally diagnosed congenital diaphragmatic hernia. AB - OBJECTIVE: The purpose of this study was to assess perinatal outcome in cases of prenatally diagnosed congenital diaphragmatic hernia and identify the prognostic value of various prenatal factors. DESIGN: Retrospective review of fetal medicine, obstetric and histopathology records in all 34 cases of congenital diaphragmatic hernia identified prenatally between 1992 and 1996 at a tertiary referral fetal medicine unit. RESULTS: Overall survival was 18%. None of those with bilateral (0/1) or right-sided (0/5) congenital diaphragmatic hernia survived, whereas 21% (6/28) of those with left-sided hernias survived. Prenatal karyotyping was performed in 71% (n = 24) and five chromosomal abnormalities were identified. The pregnancy was terminated in 15 cases (44%). In the 19 continuing pregnancies, the survival rate was 32% (6/19). In those with an isolated congenital diaphragmatic hernia, the survival rate was 38% (5/13), and in those in which the hernia was isolated and left-sided, the survival rate was 56% (5/9). In ongoing pregnancies diagnosed after 24 weeks, the survival rate was 33% (2/6) compared with 31% (4/13) in pregnancies diagnosed at < or = 24 weeks. Of those infants who had surgical repair, six out of seven (86%) survived. CONCLUSIONS: These data clearly demonstrate an improved prognosis in fetuses with an isolated left-sided congenital diaphragmatic hernia. PMID- 9744055 TI - Influence of ethnic origin on nuchal translucency screening for Down's syndrome. AB - OBJECTIVE: To determine the influence of ethnic origin on access to and equity of nuchal translucency screening for Down's syndrome in a multiethnic population. DESIGN: An observational study in a district general hospital with a large multiethnic population. SUBJECTS: 1944 women attending at a hospital antenatal clinic. MAIN OUTCOME MEASURES: First-trimester fetal nuchal translucency measurements at 10-14 weeks in women from different ethnic groups. RESULTS: The racial origin of the screened population was not significantly different from that of the population attending for initial antenatal examination. Multiple regression analysis showed a small but significant difference in nuchal translucency measurement between fetuses of different ethnic origin. CONCLUSION: Nuchal translucency screening can be effectively and equitably delivered to a multiethnic population. Although there are significant differences in nuchal translucency measurement between ethnic groups, these differences are too small to require correction when nuchal translucency is used to screen for Down's syndrome. PMID- 9744056 TI - Humerospinous distance measurements: accuracy and usefulness for predicting shoulder dystocia in delivery at term. AB - OBJECTIVE: To investigate if the humerospinous distance, as an indicator of shoulder width, could predict shoulder dystocia at term. DESIGN: Prospective cross-sectional study of a stratified reference group of healthy women and a test group of women with risk factors for shoulder dystocia. Measurements were not revealed. Outcome measures were difficulties with delivery of the shoulders and correlation with maternal body mass and birth weight. SUBJECTS: Seventy-two women with singleton pregnancies at a University Clinic: 32 women at 39-42 weeks for reference and 40 women in an at-risk test group (weight > 90 kg, weight gain > 20 kg, previous macrosomic baby, history of shoulder dystocia/difficult delivery, clinical suspicion of a large baby). Women with a breech fetus, twins and those not able to deliver vaginally were excluded. METHODS: Fetal shoulders were measured from the convergence of the cervical spinous processes at the approximate cervicothoracic vertebral junction to the medial border of the humeral head. Correlations were made with maternal body mass, birth weight, birth weight estimation, ultrasound and postnatal humerospinous measurements. RESULTS: One case of shoulder dystocia in a fetus with an average humerospinous measurement occurred in the reference group and there were no cases in the test group. There was no predictive value of a large humerospinous measurement and no correlation with maternal or fetal size. Movement of the fetal arm could change the humerospinous distance considerably, which could account for the difference between a normal and large measurement. CONCLUSION: The humerospinous distance cannot be used to predict shoulder dystocia. PMID- 9744057 TI - Cost issues surrounding the use of computerized telemedicine for obstetric ultrasonography. AB - OBJECTIVE: The purpose of this study was to describe the cost implications of converting an established videotape review network for obstetric ultrasonography to one based on telemedicine technology. DESIGN: Retrospective review of fixed and non-fixed costs associated with interpreting obstetric ultrasound examinations using both videotape and telemedicine transmission. SUBJECTS: A network of three community offices transmitting 600 obstetric ultrasound examinations per month to a central tertiary level facility. METHODS: Sonographers at the community offices record ultrasound examinations onto videotape, which are then sent by courier to a central facility for interpretation. At the completion of this videotaped examination, sonographers repeat the ultrasound scan while transmitting real-time images over a telemedicine link to the central facility. Costs associated with the videotape review technique that can be avoided by converting to telemedicine interpretation were derived and compared with the fixed and non-fixed costs associated with establishing the telemedicine network. RESULTS: For this network, the fixed costs for establishing telemedicine are $101,750. Monthly non-fixed cost savings by eliminating videotape review include $1620 to $2700 for printing still images, $1200 for courier charges and $7000 for fewer repeat ultrasound examinations. Monthly non-fixed costs for the telemedicine network are $2415. Net monthly savings in non-fixed costs for a telemedicine network are therefore $7405 to $8585, which may pay for the initial fixed costs in 12 to 14 months. CONCLUSIONS: The high cost of a telemedicine network may be offset by possible savings in non fixed costs compared with alternative systems for interpreting obstetric ultrasonography. PMID- 9744058 TI - Sonographic demonstration of nuchal cord and abnormal umbilical artery waveform heralding fetal distress. AB - Ultrasound examination of a 30-week fetus, appropriate for gestational age, revealed a nuchal cord and absence of end-diastolic velocities in the Doppler waveforms of the umbilical artery. Cardiotocography suggested impending fetal distress, clinically confirmed following expedient abdominal delivery. We suggest that Doppler velocimetry of the umbilical vessels may be useful in the assessment of viable fetuses with a sonographic diagnosis of a nuchal cord. Conversely, a nuchal cord should be considered as part of the differential diagnosis of abnormal Doppler velocimetry of the umbilical vessels. PMID- 9744059 TI - Fetal coronary and cerebral blood flow in acute fetomaternal hemorrhage. AB - A fetal heart rate tracing with absent variation and a sinusoidal pattern led to the diagnosis of acute fetomaternal hemorrhage at 29 + 2 weeks' gestation. The middle cerebral artery had increased peak blood flow velocity with reversed end diastolic flow. Fetal coronary arteries visualized by color-coded and pulsed wave Doppler sonography showed slight decrease of time-averaged maximum velocities after oxygen administration, while cerebral flows remained unchanged. After administration of 50 ml blood (pre-transfusion hematocrit < 11%) the middle cerebral artery flow normalized and coronary artery velocities decreased further until coronary blood flow could no longer be visualized within 30 min of the transfusion (post-transfusion hematocrit 27%). Evidence of successful fetal resuscitation also included normalization of the fetal heart rate tracing and resumption of fetal activity (biophysical profile score 8/10). This was also observed after a second transfusion of 38 ml blood. Deterioration from repeated fetomaternal hemorrhage led to delivery of a severely anemic neonate (cord hematocrit 7%) by Cesarean section. Postnatally, a single seizure, moderate severity respiratory distress syndrome and grade III intraventricular hemorrhage were noted. Increased peak blood flow velocity with reversed end-diastolic flow may be observed in the middle cerebral artery of fetuses with acute anemia. Correction of this phenomenon with transfusion suggests that hypovolemia and low blood viscosity are major contributing factors. Furthermore, decreasing coronary artery blood flow velocities with supplemental oxygen and blood replacement confirm functional autoregulation of the fetal coronary circulation. Observation of these acute flow changes during fetal surveillance warrants investigation for a potentially serious underlying condition. PMID- 9744060 TI - Arteriovenous fistulas of the placenta in a singleton fetus with large atrial septal defect and anomalous connection of the umbilical veins. AB - Arteriovenous fistulas of the placenta rarely occur in singleton pregnancies. This report describes the fetal and neonatal hemodynamic pattern in a singleton gestation in which multiple placental artery-to-vein anastomoses were associated with a large atrial septal defect and a single umbilical artery with an anomalous connection of the persistent right and left umbilical veins. Possible links between the extracardiac vascular malformation and the congenital heart defect are discussed. PMID- 9744061 TI - The assessment of ovarian tumor angiogenesis: what does three-dimensional power Doppler add? PMID- 9744062 TI - [QT syndrome: new diagnostic possibilities]. AB - Electrocardiographic and clinical characteristics are currently used as diagnostic criteria for the long QT-syndrome. In borderline electrocardiographic findings associated with unclear syncope, it is often difficult to ensure or exclude long QT-syndrome. Schwartz and coworkers therefore created a point system as a guide in clinical decision making. In recent years genetic diagnostics have entered the arena of long-QT assessment. Aside from new insights into the pathophysiology of the long QT-disorder, it is expected that genetic diagnostics will offer substantial help to ascertain long QT-syndrome in patients with borderline electrocardiographic and clinical findings and improve risk stratification in long-QT family members. Performing linkage analysis, coupling of autosomal-dominant congenital long QT-syndrome (Romano-Ward Syndrome) to chromosomes 11 (LQT1/11p15.5), 3 (LQT3/3p21), 7 (LQT2/7q35), and 4 (LQT4/4q25-27) was demonstrated. More recently, the disease genes in long QT-syndrome 1, 2, and 3 could be identified. Analysis of the base-pair sequence allowed detection of several different mutations in different families illustrating genetic heterogeneity. Aside from diagnostic aspects, molecular genetics may also guide pharmacological therapy by identifying the specific ion-channel disorder leading to QT-prolongation and sudden death. PMID- 9744063 TI - [Possibilities and limits of electron beam tomography]. AB - Electron beam CT, which does not require mechanical movement of an X-ray tube, has a temporal resolution which exceeds that of conventional computed tomography by a factor of about ten. Axial images of the heart can be acquired within 50 to 100 ms with a spatial resolution below 0.5 mm2 and permit precise visualization of all cardiac structures. ECG-triggered acquisition of 30 to 40 axial images (3 mm slice thickness) in a short inspiratory breathhold allows one to sample a 3 dimensional volume data set which covers the complete heart. Overall acquisition times are approximately 30 to 50 seconds. While electron beam CT in general permits a complete cardiac investigation, including morphology, function, and perfusion, the method's most important application is non-invasive imaging of the coronary arteries. Without contrast enhancement, coronary calcifications by EBCT currently constitutes the most sensitive non-invasive marker for the presence even of very early forms of coronary atherosclerosis. Intravenous injection of contrast agent during image acquisition additionally permits the selective visualization of the coronary artery lumen and detection of significant stenoses. This method is especially well suited in the follow-up of coronary interventions and coronary bypass grafting. Breathhold and movement artifacts, superposition of coronary arteries and veins, as well as severe coronary calcifications currently constitute the method's main problems. In spite of these limitations, electron beam CT has been shown to permit clinically valuable non-invasive investigations of the coronary arteries, which may be further refined as technology progresses. PMID- 9744064 TI - [Sex-specific differences in risks and management of coronary heart disease]. AB - Reports from several countries indicate that women are disadvantaged in the treatment of coronary heart disease. The aim of the present review is to examine- on the basis of current population-based data (National Health Survey [NHS], hospital diagnosis registry, mortality rates)--whether in the Federal Republic of Germany more recently a change in favor of women could be established. According to NHS results prevalence rates of hypertension, overweight, and smoking in men exceed those of women, whereas hypercholesterolaemia is equally prevalent in both sexes. From 1984/85 to 1990/91 the NHS revealed a marked decline of smoking in men, and a parallel increase in women. The prevalence of chest pain shows no gender difference, but chest pain in men is more predictive for coronary artery disease. In acute myocardial infarction [AMI] thrombolysis and PTCA are applied with equal frequency; the average length of stay in hospital is greater for women. Coronary bypass surgery and rehabilitation in hospitals are less frequently applied in women. Little is known about gender differences in psychosocial adaptation after AMI. AMI mortality rates in all age groups are lower in women, and their mean age of death is higher. However, the decline of AMI mortality since 1980 was less pronounced in women compared to men. PMID- 9744065 TI - [Acute results of ablation of coronary in-stent restenoses with eccentric excimer laser catheters]. AB - Laser catheters which run eccentrically on a guide wire were developed for maximization of luminal gain by excimer laser angioplasty (ELCA). We investigated the safety and efficacy of ELCA with these new catheters plus PTCA in patients with restenoses or occlusions in coronary stents. ELCA was performed in 57 patients (60 +/- 9 years) with stenoses in 75 stents (35 AVE Micro stents, 26 Palmaz-Schatz stents, 7 NIR stents, 7 other stents). In 44 patients eccentric 1.7 mm catheters and in 13 patients 2.0 mm catheters were used. The success of the intervention was analyzed by intravascular ultrasound (IVUS) in a subgroup of 7 patients treated with five 1.7 mm and two 2.0 mm catheters. The laser catheters could be advanced through the in-stent restenoses in 56 patients. A passage inhibition occurred in one patient with an inadequately expanded stent < 2.0 mm in diameter. ELCA reduced the diameter stenoses from 77 +/- 10% before intervention to 44 +/- 8% after treatment with the 1.7 mm catheter (n = 43) or to 34 +/- 9% after passages with the 2.0 mm catheter (P < 0.001). PTCA further reduced the diameter stenosis to 11 +/- 12% (P < 0.001). The IVUS analysis revealed a smooth ablation profile in all patients. In 4 patients creatine kinase elevations > or = 2 times normal value occurred. There was no evidence of a Q wave infarction. No dissections were observed within the stents. Outside of the stents there were dissections in 5 vessels, which required the implantation of additional stents. CONCLUSIONS: ELCA with eccentric laser catheters for treatment of in-stent restenosis is safe and effective. The incidence of complications is acceptable. PMID- 9744066 TI - [Roentgen contrast media in invasive cardiology, side effects and different indications]. AB - For invasive catheter procedures both ionic and nonionic contrast media (CM) with excellent tolerability are available. The governing practical factors for CM are X-ray opacity and biocompatibility. Tolerability of a contrast medium is governed among its physical properties by its viscosity, osmolality, and ionic concentration. In Germany the nonionic CM are currently preferred. Because of its low thrombotic complications, the ionic CM Ioxaglat is an important alternative in high risk interventions. In patients with known CM incompatibility, the prophylactic application of H1-receptor antagonists and corticosteroids allows catheterization safely without complications. In impaired renal function, hydration is the most effective prophylactic measure to be taken. PMID- 9744067 TI - [Acute vascular perforation with shunt formation in the right ventricle after percutaneous transluminal coronary angioplasty. Magnetic resonance tomography and Doppler ultrasound detection of shunt flow]. AB - A coronary artery perforation is a rare complication after percutaneous transluminal coronary angioplasty. The therapy will be determined by the hemodynamic failure of the left or right ventricle. A case of a coronary artery perforation with a shunt from the right coronary artery to the right ventricle after coronary angioplasty is reported. The shunt was detected by coronary angiography and confirmed by magnetic resonance imaging and doppler echocardiography. PMID- 9744068 TI - [Successful resuscitation of a patient with ventricular fibrillation in Bland White-Garland syndrome in adulthood. A case report]. AB - The Bland-White-Garland Syndrome represents the anomalous origin of the left coronary artery of pulmonary trunk. Only 10% of the patients reach adulthood. Clinical manifestations of the syndrome are angina, dyspnoe, ECG signs of ischemia, myocardial infarction, and death in childhood. We present the case of a 47 year old woman with Bland-White-Garland Syndrome, who was resuscitated from ventricular fibrillation. The only symptom shown in her personal history was progressive dyspnoea in the last 6 months, though mitral insufficiency was known since childhood. On echocardiographic examination, she showed an anterolateral infarction and a mitral insufficiency II. As operation procedure, the ligation of the left main coronary artery and bypass surgery with a left internal mammarian graft to the left descending branch of the left coronary artery was chosen. The mechanism of onset of ventricular tachycardia in our patient is not known. Three pathophysiological mechanisms may be possible: (1) local ischemia caused by the shunt, (2) a reentry circuit in the border zone of myocardial infarction, (3) electrical instability caused by endocardial fibrosis. As local ischemia and reentry circuit were widely excluded, only endocardial fibrosis could induce further ventricular arrhythmia. We therefore intended to implant an AICD to have the most possible safety for our patient. But this, postoperatively was refused by the patient. In analogy to Coronary Artery Disease, the risk for sudden cardiac death postoperatively may be due to three factors: (1) presence of a reentrant circuit, (2) LV-function below 40%, and (3) presence of endocardial fibrosis. Our patient showed a low risk for sudden cardiac death. On electrophysiological study, no ventricular tachycardia could be induced in our patient, indicating the absence of a reentry circuit. LV function exceeded more than 40%. In Holter ECG, only few ventricular premature beats could be registrated, indicating a low risk for sudden cardiac death in the presence of endocardial fibrosis. In the follow-up of fourteen months, the patient remained free from arrhythmic events. PMID- 9744069 TI - Children with parents in prison: child welfare policy, program, and practice issues. AB - As the rate at which adults are being incarcerated in the United States escalates, child welfare professionals are encountering growing numbers of children who have parents in prison. Current estimates indicate that as many as 1.5 million children have an incarcerated parent; many thousands of others have experienced the incarceration of a parent at some point in their lives. These vulnerable children face unique difficulties, and their growing numbers and special needs demand attention. Challenges facing the child welfare system as it attempts to work with this population are explored. PMID- 9744070 TI - Supporting families and children of mothers in jail: an integrated child welfare and criminal justice strategy. AB - Charged with the responsibility for protecting children and preserving families, the child welfare system needs to pay special attention to women in jail and their families. Jails provide an opportunity to reach families early in the criminal justice process. This article explores why working with women in jail and their children is within the scope of the child welfare system's mandate; describes the pressures on the criminal justice and child welfare systems that prevent either from working effectively with these families; and suggests a collaborative strategy for working more effectively with mothers in jail and their children. PMID- 9744071 TI - Charting a course: meeting the challenge of permanency planning for children with incarcerated mothers. AB - Case workers involved in permanency planning for children whose mothers are incarcerated must assess the family's strengths, the mother's capacity to assume parental responsibilities, and the integrity of the parent-child relationship; and address concerns regarding the short- and long-term effects of the children's socioemotional dislocation and the merits of their retaining a relationship with their mothers. At the same time, correctional policies and practices delineate the nature and extent of contacts with the mother and the mother's access to rehabilitative programs. Agency guidelines, practice tools, and advocacy initiatives must be developed to help practitioners meet these challenges. An initial review of the Adoption and Safe Families Act suggests the need for close monitoring of the impact of its mandates to shorten the time for moving children toward permanency and its weakening of the expectation for "reasonable efforts" to be made to reunify families. PMID- 9744072 TI - In whose best interest? The impact of changing public policy on relatives caring for children with incarcerated parents. AB - Changes in criminal justice policy have resulted in the incarceration of an unprecedented number of parents. Consequently, more children than ever before are living with grandparents and other relatives while their parents are imprisoned. Historically, child welfare and criminal justice policy have been treated as distinct and unrelated areas of policy. This article discusses the interdependence of criminal justice policy, welfare reform legislation, and kinship foster care, and the impact of this interdependence on children whose parents are in jail or in prison. PMID- 9744073 TI - Permanency planning in the context of parental incarceration: legal issues and recommendations. AB - Parental incarceration is a growing problem that is not accommodated for in the traditional, time-driven model of permanency planning. The trend in both law and policy regarding children in out-of-home care is toward early termination of parental rights and placement for adoption. The federal Adoption and Safe Families Act of 1997 places additional pressure on state agencies to continue this trend. Child welfare agencies need to recognize the importance of maintaining parent-child relationships, even when a parent is incarcerated, and must develop creative approaches for dealing with the unique challenges presented by parental incarceration. The author, an advocate for incarcerated parents, offers recommendations for permanency planning in the context of parental incarceration. PMID- 9744074 TI - Girl Scouts Beyond Bars: facilitating parent-child contact in correctional settings. AB - Distant prison locations, inconvenient visiting schedules, and the negative effects of a mother's imprisonment on her children often complicate the child welfare professional's work with the children of incarcerated mothers. Enhanced prison visiting programs offer a mechanism to support the mother-child relationship, facilitate reunification efforts, and assist with permanency planning. The article discusses one such program, Girl Scouts Beyond Bars, in relation to the problem of mother-child separation via imprisonment; traditional visiting practices; and the issues confronting child welfare professionals serving the children of imprisoned mothers. PMID- 9744075 TI - Earning trust from youths with none to spare. AB - The Center for Community Alternatives (CCA) operates an array of programs that primarily serve students whose parents are incarcerated. The authors explore outreach and programmatic approaches and discuss the benefits of a holistic, multifaceted, open approach to identifying, assessing, and meeting the needs of adolescent children of incarcerated parents. The article concludes with policy recommendations regarding further development of programs targeting this population. PMID- 9744076 TI - The forgotten parent: understanding the forces that influence incarcerated fathers' relationships with their children. AB - Fathers who are in prisons and jails are not just convicts--they are parents as well. The family roles and responsibilities of incarcerated fathers, however, are seldom the focus of institutional policies, scholarly research, or child welfare services. This article examines the issues that must be addressed in designing policies and providing services that promote the maintenance of parent-child bonds and responsible parenting when fathers are incarcerated. It describes the family roles and structures of men in prison and looks at differences between public perceptions and the real-life experiences of prisoner parents. The ways in which correctional policies and child welfare practices influence and shape fathers' parenting abilities and father-child relationships are discussed, and strategies for creating a supportive environment for fathers and their children and families are proposed. PMID- 9744077 TI - The p21Rac/Cdc42-activated kinases (PAKs). AB - The p21-activated kinases (PAKs) are mammalian Rac/Cdc42-associated serine/threonine protein kinases. They contain diverse structural regulatory elements that allow them not only to participate as effectors in signaling processes initiated by activated GTPases but also in signal transduction events mediated by Src3 homology domains (SH3) or caspase-mediated proteolytic cleavage. The biological functions of PAK protein kinases result from the interplay of N- and C-terminal-mediated protein-protein interactions and signals derived from phosphorylation of downstream substrates. The potential regulation of microbial killing, stress responses, apoptosis, and cell motility by PAKs suggest it may be a therapeutically useful target in a number of disease states. PMID- 9744078 TI - Carnosine, a protective, anti-ageing peptide? AB - Carnosine (beta-alanyl-L-histidine) has protective functions additional to anti oxidant and free-radical scavenging roles. It extends cultured human fibroblast life-span, kills transformed cells, protects cells against aldehydes and an amyloid peptide fragment and inhibits, in vitro, protein glycation (formation of cross-links, carbonyl groups and AGEs) and DNA/protein cross-linking. Carnosine is an aldehyde scavenger, a likely lipofuscin (age pigment) precursor and possible modulator of diabetic complications, atherosclerosis and Alzheimer's disease. PMID- 9744079 TI - Lung smooth muscle differentiation. AB - The vascular and visceral smooth muscle tissues of the lung perform a number of tasks that are critical to pulmonary function. Smooth muscle function often is compromised as a result of lung disease. Though a great deal is known about regulation of smooth muscle cell replication and cell and tissue contractility, much less is understood regarding the phenotype of the contractile protein machinery of lung smooth muscle cells. This review focuses on the expression of cytoskeletal and contractile proteins of lung vascular and airway smooth muscle cells during development, in the adult and during vascular and airway remodeling. Emphasis is placed on the expression of the heavy chain of smooth muscle myosin, as well as the regulation of its gene. Important areas for future research are discussed. PMID- 9744080 TI - Metabolic effects of thiopurine derivatives against human CCRF-CEM leukaemia cells. AB - BACKGROUND and aims. To compare the metabolic effects induced by the anticancer drugs, 6-mercaptopurine (6-MP), 6-thioguanine (6-TG) and 6-methylmercaptopurine riboside (MMPR), which may inhibit the de novo biosynthesis of purine nucleotides or be mis-incorporated into DNA or RNA. METHODS: Leukaemia cells were grown in culture, exposed to a thiopurine and cell extracts were analyzed for NTPs, dNTPs, drug metabolites and P-Rib-PP. RESULTS: In leukaemia cells, 6-MP was converted to MPR-MP, thio-XMP, thio-GMP, thio-GDP and thio-GTP. Metabolites of 6-TG included thio-XMP, thio-GMP, thio-GDP and thio-GTP, while MMPR-MP was the only major metabolite of MMPR, MMPR (25 microM, 4 h) induced a 16-fold increase in P-Rib-PP and 6-MP (25 microM, 4 h) induced a delayed 5.2-fold increase. MPR-MP, thio-GMP and MMPR-MP are inhibitors of amido phosphoribosyltransferase from leukaemia cells with Ki values of 114 +/- 7.10 microM, 6.20 +/- 2.10 microM and 3.09 +/- 0.30 microM, respectively. CONCLUSION: The nucleoside-5'-monophosphate derivatives of the 3 thiopurines inhibit amido phosphoribosyltransferase in growing leukaemia cells but there is also an initial inhibition of the further conversion of IMP in the pathway. In growing cells, MMPR acts solely as an inhibitor of de novo purine biosynthesis while 6-TG and to a lesser extent, 6-MP, are converted to significant concentrations of di- and tri-phosphate derivatives which may have other mechanisms of cytotoxicity. PMID- 9744081 TI - Purification of a novel muscle cell growth factor S-myotrophin from porcine skeletal muscle. AB - A comparison of muscle weight between denerved and control rabbit hind legs revealed that a water-soluble 12 kDa substance was reduced in atrophied muscles after denervation. We hypothesised that a water-soluble growth factor exists which mediates a signal from motor nerves to muscles. To isolate this factor we modified the purification procedures of Sen et al. [S. Sen, G. Kundu, N. Mekhail, J. Castel, K. Misono, B. Healy, Myotrophin: purification of a novel peptide from spontaneously hypertensive rat heart that influences myocardial growth, J. Biol. Chem. 265 (1990) 16635-16643.], who originally purified a water-soluble growth factor from cardiac muscles. Four additional purification steps were added to the method. Using this technique, a novel muscle cell growth factor, named s myotrophin, was purified from porcine skeletal muscle (M. longissimus thoracis). Purified s-myotrophin appeared as a single band (12 kDa) on SDS-PAGE and had a strong growth promoting activity (increase of protein synthesis) of cultured primary skeletal muscle cells. Almost no loss of growth promoting activity was observed after trypsin and chymotrypsin digestion. No fragmentation of s myotrophin was observed after exposure to lysylendopeptidase, thermolysin, trypsin and chymotrypsin. Crude preparation of this molecule could be detected by periodic acid/Schiff (PAS) staining. Deglycosylation of s-myotrophin produced a smaller molecule having an approximately 7 kDa mass. These data indicate a novel 12 kDa protein has been isolated which has growth promoting properties on skeletal muscle cells. PMID- 9744082 TI - Connective tissue growth factor: a novel regulator of mucosal repair and fibrosis in inflammatory bowel disease? AB - Inflammatory bowel disease (IBD) is a multifactorial disorder which is characterized by massive damage of the epithelium and the underlying mesenchyme of the intestine. Due to the potent effect of connective tissue growth factor (CTGF) on fibroblast proliferation and connective tissue deposition we speculated about a possible role of this mitogen in IBD. Here we demonstrate a strikingly increased expression of CTGF mRNA in surgical specimens of patients suffering from two forms of IBD, Crohn's disease and ulcerative colitis. In most specimens, the levels of CTGF mRNA correlated with the degree of inflammation as assessed by histological analysis of adjacent tissue samples and by expression analysis of the pro-inflammatory cytokine interleukin-1 beta. However, areas of little inflammation which were characterized by severe fibrosis also revealed high levels of CTGF mRNA. Expression of transforming growth factor beta-1 (TGF-beta 1), the only known inducer of CTGF so far, as well as of the CTGF target genes collagen I alpha 1, fibronectin and integrin alpha 5 revealed a strong correlation with the expression of CTGF. These data suggest a prominent role of CTGF in the repair of mucosal injury in IBD and in the aberrant deposition of extracellular matrix leading to fibrosis and stenosis, one major complication in IBD, especially in Crohn's disease. PMID- 9744083 TI - Internalization study using EDTA-prepared hepatocytes for receptor-mediated endocytosis of haemoglobin-haptoglobin complex. AB - We have demonstrated the internalization of haemoglobin-haptoglobin (Hb-Hp) complex using rat hepatocytes prepared by EDTA perfusion, followed by Percoll. The isolated hepatocytes exhibited a saturation curve of the binding of fluorescein isothiocyanate-labelled haemoglobin-haptoglobin complex (FITC-Hb-Hp. Furthermore, competition between the binding of FITC-Hb-Hp and unlabelled Hb to the hepatocytes, was observed. The cells exhibited approximately 9 x 10(4) 'high affinity sites' (Kd approximately 1.2 microM) for the Hb-Hp complex. The data in toto suggest the presence of only one type of receptor i.e. the high affinity receptor (in both affinity and number of sites per cell). The results were similar to those obtained from rat hepatocytes prepared by collagenase digestion [1]. In order to verify whether EDTA-prepared hepatocytes could be used for the study of receptor-mediated endocytosis, the internalization of pre-bound Hb-Hp in the isolated hepatocytes was assessed by two methods. First, acid-insensitive FITC-Hb-Hp time-dependently increased following incubation at 37 degrees C. Secondly, Hb-Hp became inaccessible to the exogenous FITC-anti-haemoglobin antibody. These processes were dependent on ATP, but independent of Ca2+ and stimulated by GTP. The results demonstrate that the receptor-mediated endocytosis of Hb-Hp occurred in the EDTA-prepared hepatocytes. PMID- 9744084 TI - Relationship between the catalytic sites of human bifunctional IMP synthase. AB - BACKGROUND AND AIMS: The bifunctional enzyme, IMP synthase, contains 5 aminoimidazole-4-carboxamide ribotide (AICAR) transformylase and IMP cyclohydrolase activities and catalyses the ninth and tenth reactions of the pathway for de novo biosynthesis of purine nucleotides (AICAR-->FAICAR-->IMP). The spatial relationship between the two active sites on IMP synthase has been investigated along with the possibility that the intermediate, FAICAR, may be channelled between the two sites. METHODS: The two catalytic activities and the overall reaction (AICAR-->FAICAR-->IMP) were assayed using 3H-labelled AICAR or FAICAR with isolation of the reaction products by thin-layer chromatography. RESULTS: Inhibition constants for the interactions of six purine nucleoside 5' monophosphate derivatives with AICAR transformylase and IMP cyclohydrolase were 24- to 820-fold higher for the transformylase. N-ethylmaleimide inactivated IMP cyclohydrolase but not AICAR transformylase. The rate of IMP synthesis from AICAR was consistent with a high local concentration of FAICAR at the cyclohydrolase site but addition of exogenous unlabelled FAICAR reduced the amount of [3H]AICAR formed from [3H]AICAR indicating that the channelling of FAICAR was not absolute. CONCLUSION: The AICAR transformylase and IMP cyclohydrolase active sites of IMP synthase are distinct but sufficiently close for the FAICAR produced by a transformylase site to be preferentially utilized as a substrate by a cyclohydrolase site on the same molecule if dimeric, bifunctional IMP synthase. PMID- 9744085 TI - Effects of niacin deficiency on the levels of soluble proteins and enzyme activities in various tissues of Japanese quail. AB - The effects of niacin deficiency on the levels of soluble proteins and enzyme activities of Japanese quail have been investigated. SDS-polyacrylamide gel electrophoresis revealed that in the pectoral muscle the soluble proteins with molecular masses corresponding to 181, 128, 93, 76, 72, 62, 56, 43, 41, 28 and 20 kDa were present in lower amounts but those of 60, 50 and 37 kDa were present in higher amounts. In the heart the soluble proteins with a molecular mass of 181 kDa were present in lower amounts and in the brain those of 43 kDa were present in lower amounts but those of 221 kDa were present in higher amounts. In the intestine the soluble proteins with molecular masses corresponding to 181, 102, 83, 74, 72, 44 and 40 kDa were present in lower amounts but those of 41 kDa and 18 kDa were present in higher amounts. There was a marked reduction in the level of NAD and NADPH in the pectoral muscle of niacin deficient quail but not in other tissues. The specific activity of glyceraldehyde-3-phosphate dehydrogenase decreased markedly both in the liver and pectoral muscle of niacin deficient quail whereas that of 6-phosphogluconate dehydrogenase and malic enzyme decreased markedly in the liver or pectoral muscle, respectively. In contrast, the specific activity of acetylcholinesterase and carboxypeptidase increased markedly in the liver or the pectoral muscle, respectively. The results suggest that a severe niacin deficiency exerted specific effects on levels of some soluble proteins particularly in the pectoral muscle and intestine and on activities of certain enzymes in the liver and the pectoral muscle. PMID- 9744086 TI - The significance of locating and filling the canal isthmus in multiple root canal systems. A scanning electron microscopy study of the mesiobuccal root of maxillary first permanent molars. AB - The mesiobuccal (MB) roots of 50 randomly selected maxillary first permanent molars were examined to evaluate the different configurations of canal isthmus and their incidences. Sections of the roots at 3.4, and 5 mm from the apex were prepared, acid etched, washed and dried. The apical side of each section was sputter coated with gold, examined by a Hitachi S-2500 scanning electron microscope and photographed. 36% of the mesiobuccal roots had one canal, whereas 64% had two canals. In the roots with two canals 31.25% contained either a complete isthmus, or accessory canals between the two main canals 31.25% showed partial isthmus formation. The significance of the different configurations is discussed. Our present findings indicate that the mesiobuccal roots of maxillary first permanent molars exhibit complex anatomy. Prudent judgement is essential in the canal isthmus preparation so that operators must provide meticulous skill and care to the patients. PMID- 9744087 TI - Modification of Unicryl composition for rapid polymerization at low temperature without alteration of immunocytochemical sensitivity. AB - We introduced a slight modification of Unicryl for fast polymerization at low temperature (in the range-20 degree C to-35 degree C) without modification of both physical properties and immunocytochemical yield. An UV initiator benzoin ethyl ether (BEE) is added at 0.05% concentration in neat resin during infiltration and final polymerization at-35 degree C. Immunogold labeling of amylase of mouse pancreas embedded in standard conditions recommended by the manufacturer or with our modification were compared by counting colloidal gold particles over zymogen granules using MACS, a specialized software dedicated to this operation. We noticed that addition of 0.05% BEE in Unicryl reduces the curing time to 16 h, which can be extended up to a maximum of 48 h at-35 degree C without noticeable modification of ultrastructural morphology, physical properties and immunocytochemical yield. PMID- 9744088 TI - Optimization of phosphorus localization by EFTEM of nucleic acid containing structures. AB - Energy Filtered Transmission Electron Microscopy (EFTEM) has been used to study nucleic acids localization in unstained thin sections of virus-infected cells. For this purpose, phosphorus maps (P-maps) have been obtained by applying the N windows Egerton model for background subtraction from data acquired by a non dedicated TEM Jeol 1200EXII equipped with a post-column PEELS Gatan 666-9000 and a Gatan Image Filter (GIF-100). To prevent possible errors in the evaluation of elemental maps and thus incorrect nucleic acid localization, we have studied different regions of swine testis (ST) cells with similar local density containing either high concentration of nucleic acids (condensed chromatin and ribosomes) or a very low concentration (mitochondria). Special care was taken to optimize the sample preparation conditions to avoid as much as possible the traditional artifacts derived from this source. Selection of the best set of pre edge images for background fitting was also considered in order to produce "true P-maps". A new software for interactive processing of images series has been applied to estimate this set. Multivariate Statistical Analysis was used as a filtering tool to separate the "useful information" present in the inelastic image series (characteristic signal) from the "non-useful information" (noise and acquisition artifacts). The reconstitution of the original image series preserving mainly the useful information allowed the computation of P-maps with improved signal-to-noise ratio (SNR). This methodology has been applied to study the RNA content of maturation intermediate coronavirus particles found inside infected cells. PMID- 9744089 TI - Aspects of the structure and assembly of desmosomes. AB - Desmosomes are found principally in epithelial cells and consist of disc-like plaques, the extracellular face of which is paired with that of a neighbouring cell. There is increasing evidence that desmosomes are adhesive structures, and that two types of desmosomal glycoproteins, the desmogleins (Dsg) and desmocollins (Dsc) both Ca(2+)-binding cadherin-like molecules, perform this role in adhesion through interaction of their extracellular domains. A number of isoforms of Dsg and Dsc are present in specific tissues. The cytoplasmic side of the plaque is attached to intermediate filaments through desmoplakin, a major plaque protein. Also associated with desmosomes are plakoglobin and beta-catenin, suggesting that the adhesive function of desmosomes might be mediated by signal transduction. Formation of desmosomes can be studied by growing epithelial cells in low-Ca2+ medium (LCM, < 0.1 mM), where desmosomal proteins are either synthesized but not assembled, or form partially assembled but unstable half desmosomes. Addition of Ca2+ (to about 2mM) initiates cell contact and, in the case of half-desmosomes, leads to stabilization by incorporation into membranes and formation of typical paired structures. In cases where such pre-assembled structures are not formed, recruitment of desmosomal proteins appears to occur by vesicular transport of desmocollins and desmogleins to the cell surface, where association is made with plakoglobin and later, with desmoplakin. Although much remains to be learned of the assembly process, specific interacting domains of the molecular components are being recognized. Desmosome assembly is part of a coordinated pattern of junction formation which accompanies the establishment of cell polarity, resulting in differentiation of apical and basolateral cell surfaces. Desmosomes are now being regarded, not as static and inert structures, but as membrane specializations linked to systems involved in cell-cell communication as well as adhesion. PMID- 9744090 TI - NADPH is a specific inhibitor of protein import into glyoxysomes. AB - We have studied the import of proteins into glyoxysomes in vitro and show that this process is specifically inhibited by NADPH. NADPH affects both binding and translocation of proteins into glyoxysomes, and inhibition is determined by the ratio of NADP+ to NADPH. The site of action of NADPH is most likely within the glyoxysome because (1) pretreatment of glyoxysomes with NADPH, followed by re isolation of the organelles prior to the import assay, resulted in inhibition of import that could be restored by the addition of NADP+; (2) low concentrations of NADPH inhibited binding of proteins to broken glyoxysome membranes. The sensitivity of protein import to inhibition by NADPH declines as glyoxysomes are converted to leaf-type peroxisomes. A model is proposed that speculates on a possible role for NADPH in regulating protein import into plant peroxisomes. PMID- 9744091 TI - Domains of the TMV movement protein involved in subcellular localization. AB - To identify and map functionally important regions of the tobacco mosaic virus movement protein, deletions of three amino acids were introduced at intervals of 10 amino acids throughout the protein. Mutations located between amino acids 1 and 160 abolished the capacity of the protein to transport virus from cell to cell, while some of the mutations in the C-terminal third of the protein permitted function. Despite extensive tests, no examples were found of intermolecular complementation between mutants, suggesting that function requires each movement protein molecule to be fully competent. Many of the mutants were fused to green fluorescent protein, and their subcellular localizations were determined by fluorescence microscopy in infected plants and protoplasts. Most mutants lost the ability to accumulate in one or more of the multiple subcellular sites targeted by wild-type movement protein, suggesting that specific functional domains were disrupted. The order in which accumulation at subcellular sites occurs during infection does not represent a targeting pathway. Association of the movement protein with microtubules or with plasmodesmata can occur in the absence of other associations. The region of the protein around amino acids 9-11 may be involved in targeting the protein to cortical bodies (probably associated with the endoplasmic reticulum) and to plasmodesmata. The region around residues 49-51 may be involved in co-alignment of the protein with microtubules. The region around residues 88-101 appears to play a role in targeting to both the cortical bodies and microtubules. Thus, the movement protein contains independently functional domains. PMID- 9744092 TI - A xyloglucan oligosaccharide-active, transglycosylating beta-D-glucosidase from the cotyledons of nasturtium (Tropaeolum majus L) seedlings--purification, properties and characterization of a cDNA clone. AB - A beta-D-glucosidase has been purified to apparent homogeneity from the cotyledons of germinated nasturtium (Tropaeolum majus L.) seedlings during the mobilization of the xyloglucan stored in the cotyledonary cell walls. The purified protein (Mr 76, 000; a glycoprotein; pl > 9.5; apparent pH optimum 4.5; temperature optimum 30 degrees C) catalysed the hydrolysis of p-nitrophenyl-beta D-glucopyranoside, cello-oligosaccharides, beta-linked glucose disaccharides, and certain xyloglucan oligosaccharides. Glucose disaccharides with different linkages were hydrolysed at different rates [(1-->3) > (1-->4) > (1-->2) > (1- >6)] with significant transglycosylation occurring in the early stages of the reaction. Cello-oligosaccharide hydrolysis was also accompanied by extensive transglycosylation to give transitory accumulations of higher oligosaccharides. At least some of the glycosyl linkages formed during transglycosylation were (1- >6)-beta. Xyloglucan oligosaccharides xylose-substituted at the non-reducing terminal glucose residue (XXXG, XXLG, XLXG and XLLG, where G is an unsubstituted glucose residue, X is a xylose-substituted glucose residue, and L is a galactosylxylose-substituted glucose residue) were not hydrolysed. Some xyloglucan oligosaccharides with an unsubstituted non-reducing terminal glucose residue (GXXG, GXLG and GXG) were hydrolysed, but others (GLXG and GLLG) were not. This indicated steric hindrance by L but not X substitution at the glucose residue next to the one at the non-reducing end of the oligosaccharide. Hydrolysis of xyloglucan oligosaccharides was not accompanied by transglycosylation. Natural xyloglucan subunit oligosaccharides (XXXG, XXLG, XLXG, XLLG) were totally degraded to their monosaccharide components when treated with nasturtium beta-D-galactosidase. (Edwards et al (1988) J. Biol. Chem. 263, 4333-4337), followed by alternations of nasturtium xyloglucan-specific alpha xylosidase (Fanutti et al (1991) Planta 184, 137-147) and this enzyme. Several extensively overlapping cDNA clones were obtained by RT-PCR and by screening cDNA libraries. A composite, full-length DNA had an open reading frame of 1962 bp, encoding a polypeptide of 654 amino acids, including all N-terminal and internal sequences obtained from the purified beta-glucosidase protein, and a motif resembling plant signal sequences thought to direct proteins to the cell wall. Database searches revealed homology with beta-glucosidases from several sources (plant, bacteria, yeast), notably with glycosylhydrolases of 'Family 3', according to the classification of Henrissat (Henrissat (1991) Biochem. J. 280, 309-316). There was strong sequence homology with a beta-glucan exo-hydrolase from barley (Hrmova et al. (1996) J. Biol. Chem. 271, 5277-5286). The nasturtium beta-glucosidase is ascribed a role in xyloglucan mobilization, and its interaction with the alpha-xylosidase and the beta-galactosidase is modelled. PMID- 9744093 TI - Identification of a novel delta 6-acyl-group desaturase by targeted gene disruption in Physcomitrella patens. AB - The moss Physcomitrella patens contains high levels of arachidonic acid. For its synthesis from linoleic acid by desaturation and elongation, novel delta 5- and delta 6-desaturases are required. To isolate one of these, PCR-based cloning was used, and resulted in the isolation of a full-length cDNA coding for a putatively new desaturase. The deduced amino acid sequence has three domains: a N-terminal segment of about 100 amino acids, with no similarity to any sequence in the data banks, followed by a cytochrome b5-related region and a C-terminal sequence with low similarity (27% identify) to acyl-lipid desaturases. To elucidate the function of this protein, we disrupted its gene by transforming P. patens with the corresponding linear genomic sequence, into which a positive selection marker had been inserted. The molecular analysis of five transformed lines showed that the selection cartridge had been inserted into the corresponding genomic locus of all five lines. The gene disruption resulted in a dramatic alteration of the fatty acid pattern in the knockout plants. The large increase in linoleic acid and the concomitant disappearance of gamma-linolenic and arachidonic acid in all knockout lines suggested that the new cDNA coded for a delta 6-desaturase. This was confirmed by expression of the cDNA in yeast and analysis of the resultant fatty acids by GC-MS. Only the transformed yeast cells were able to introduce a further double bond into the delta 6-position of unsaturated fatty acids. To our knowledge, this is the first report of a successful gene disruption in a multicellular plant resulting in a specific biochemical phenotype. PMID- 9744094 TI - Promoters of nuclear-encoded respiratory chain complex I genes from Arabidopsis thaliana contain a region essential for anther/pollen-specific expression. AB - Regulatory promoter regions responsible for the enhanced expression in anthers and pollen are defined in detail for three nuclear encoded mitochondrial Complex I (nCl) genes from Arabidopsis thaliana. Specific regulatory elements were found conserved in the 5' upstream regions between three different genes encoding the 22 kDa (PSST), 55 kDa NADH binding (55 kDa) and 28 kDa (TYKY) subunits, respectively. Northern blot analysis and transgenic Arabidopsis plants carrying progressive deletions of the promoters fused to the beta-glucuronidase (GUS) reporter gene by histochemical and fluorimetric methods showed that all three promoters drive enhanced expression of GUS specifically in anther tissues and in pollen grains. In at least two of these promoters the -200/-100 regions actively convey the pollen/anther-specific expression in gain of function experiments using CaMV 35S as a minimal promoter. These nCl promoters thus contain a specific regulatory region responding to the physiological demands on mitochondrial function during pollen maturation. Pollen-specific motifs located in these regions appear to consist of as little as seven nucleotides in the respective promoter context. PMID- 9744095 TI - Photomorphogenic development of the Arabidopsis shy2-1D mutation and its interaction with phytochromes in darkness. AB - We previously reported a photomorphogenic mutation of Arabidopsis thaliana, shy2 1D, as a dominant suppressor of a hy2 mutation. Here, we report that shy2-1D confers various photo-responsive phenotypes in darkness and the dark phenotypes of the mutant are affected by phytochrome deficiency. Dark-grown seedlings of the mutant developed several photomorphogenic characteristics such as short hypocotyls, cotyledon expansion and opening, and partial differentiation of plastids. When grown further in darkness, the mutant plant underwent most of the developmental stages of a light-grown wild-type plant, including development of foliar leaves, an inflorescence stem with cauline leaves, and floral organs. In addition, two light-inducible genes, the nuclear-encoded CAB and the plastid encoded PSBA genes, were highly expressed in the dark-grown mutant seedlings. Furthermore, reduced gravitropism, a phytochrome-modulated response, was observed in the mutant hypocotyl in darkness. Thus, shy2-1D is one of the most pleiotropic photomorphogenic mutations identified so far. The results indicate that SHY2 may be a key component regulating photomorphogenesis in Arabidopsis. Surprisingly, double mutants of the shy2-1D mutant with the phytochrome-deficient mutants hy2, hy3(phyB-1) and fre1-1(phyA-201) showed reduced photomorphogenic response in darkness with a longer hypocotyl, a longer inflorescence stem, and a lower level expression of the CAB gene than the shy2-1D single mutant. These results showed that phytochromes function in darkness in the shy2-1D mutant background. The implications of these results are discussed. PMID- 9744097 TI - Tetrad pollen formation in quartet mutants of Arabidopsis thaliana is associated with persistence of pectic polysaccharides of the pollen mother cell wall. AB - The quartet (qrt) mutants of Arabidopsis thaliana produce tetrad pollen in which microspores fail to separate during pollen development. Because the amount of callose deposition between microspores is correlated with tetrad pollen formation in other species, and because pectin is implicated as playing a role in cell adhesion, these cell-wall components in wild-type and mutant anthers were visualized by immunofluorescence microscopy at different stages of microsporogenesis. In wild-type, callose was detected around the pollen mother cell at the onset of meiosis and around the microspores during the tetrad stage. Microspores were released into the anther locule at the stage where callose was no longer detected. Deposition and degradation of callose during tetrad pollen formation in qrt1 and qrt2 mutants were indistinguishable from those in wild type. Enzymatic removal of callose from wild-type microspores at the tetrad stage did not release the microspores, suggesting that callose removal is not sufficient to disperse the microspores in wild-type. Pectic components were detected in the primary wall of the pollen mother cell. This wall surrounded the callosic wall around the pollen mother cell and the microspores during the tetrad stage. In wild-type, pectic components of this wall were no longer detectable at the time of microspore release. However, in qrt1 and qrt2 mutants, pectic components of this wall persisted after callose degradation. This result suggests that failure of pectin degradation in the pollen mother cell wall is associated with tetrad pollen formation in qrt mutants, and indicates that QRT1 and QRT2 may be required for cell type-specific pectin degradation to separate microspores. PMID- 9744096 TI - Combinatorial interaction of light-responsive elements plays a critical role in determining the response characteristics of light-regulated promoters in Arabidopsis. AB - We have studied the roles of PhyA, PhyB and CRY1 photoreceptors and the downstream light-signaling components, COP1 and DET1, in mediating high irradiance light-controlled activity of promoters containing synthetic light responsive elements (LRE). Promoters with paired LREs were able to respond to a wide spectrum of light through multiple photoreceptors, while the light-inducible single LRE promoters primarily responded to a specific wavelength of light. In addition, our results indicate that Cry1 is involved in PhyB-mediated red-light induction of the G-GATA/NOS101 promoter, and that both Cry1 and PhyB are required for effective repression of the GT1/NOS101 promoter by red or blue light. An interaction between PhyA and PhyB in mediating GT1-GATA/NOS101 promoter light activation was also observed. Furthermore, our data indicate that COP1 and DET1 exert negative control in the dark only on paired LRE promoters but not single LRE promoters. From these results, we conclude that the combinatorial interaction of LREs is essential in determining the ability of light-responsive promoters to be modulated by crucial cellular regulators and to respond to diverse light environments. PMID- 9744098 TI - Low-oxygen stress and water deficit induce cytosolic pyruvate orthophosphate dikinase (PPDK) expression in roots of rice, a C3 plant. AB - Pyruvate orthophosphate dikinase (PPDK) is known for its role in C4 photosynthesis but has no established function in C3 plants. Abscisic acid, PEG and submergence were found to markedly induce a protein of about 97 kDa, identified by microsequencing as PPDK, in rice roots (C3). The rice genome was found to contain two ppdk loci, osppdka and osppdkb. We isolated osppdka cDNA, which encodes a cytosolic rice PPDK isoform of 96.6 kDa, that corresponded to the ABA-induced protein from roots. Western blot analysis showed a PPDK induction in roots of rice seedlings during gradual drying, cold, high salt and mannitol treatment, indicating a water deficit response. PPDK was also induced in the roots and sheath of submerged rice seedlings, and in etiolated rice seedlings exposed to an oxygen-free N2 atmosphere, which indicated a low-oxygen stress response. None of the stress treatments induced PPDK protein accumulation in the lamina of green rice seedlings. Ppdk transcripts were found to accumulate in roots of submerged seedlings, concomitant with the induction of alcohol dehydrogenase 1. Low-oxygen stress triggered an increase in PPDK activity in roots and etiolated rice seedlings, accompanied by increases in phosphoenolpyruvate carboxylase and malate dehydrogenase activities. The results indicate that cytosolic PPDK is involved in a metabolic response to water deficit and low-oxygen stress in rice, an anoxia-tolerant species. PMID- 9744099 TI - Phytobilin biosynthesis: cloning and expression of a gene encoding soluble ferredoxin-dependent heme oxygenase from Synechocystis sp. PCC 6803. AB - The phytobilin chromophores of phycobiliproteins and phytochromes are biosynthesized from heme in a pathway that begins with the opening of the tetrapyrrole macrocycle of protoheme to form biliverdin IX alpha, in a reaction catalyzed by heme oxygenase. A gene containing an open reading frame with a predicted polypeptide that has a sequence similar to that of a conserved region of animal microsomal heme oxygenases was identified in the published genomic sequence of Synechocystis sp. PCC 6803. This gene, named ho1, was cloned and expressed in Escherichia coli under the control of the lacZ promoter. Cells expressing the gene became green colored due to the accumulation of biliverdin IX alpha. The size of the expressed protein was equal to the predicted size of the Synechocystis gene product, named HO1. Heme oxygenase activity was assayed in incubations containing extract of transformed E. coli cells. Incubations containing extract of induced cells, but not those containing extract of uninduced cells, had ferredoxin-dependent heme oxygenase activity. With mesoheme as the substrate, the reaction product was identified as mesobiliverdin IX alpha by spectrophotometry and reverse-phase HPLC. Heme oxygenase activity was not sedimented by centrifugation at 100, 000 g. Expression of HO1 increased several fold during incubation of the cells for 72 h in iron-deficient medium. PMID- 9744100 TI - Characterization of exon skipping mutants of the COP1 gene from Arabidopsis. AB - The removal of introns from pre-mRNA requires accurate recognition and selection of the intron splice sites. Mutations which alter splice site selection and which lead to skipping of specific exons are indicative of intron/exon recognition mechanisms involving an exon definition process. In this paper, three independent mutants to the COP1 gene in Arabidopsis which show exon skipping were identified and the mutations which alter the normal splicing pattern were characterized. The mutation in cop1-1 was a G-->A change 4 nt upstream from the 3' splice site of intron 5, while the mutation in cop1-2 was a G-->A at the first nucleotide of intron 6, abolishing the conserved G within the 5' splice site consensus. The effect of these mutations was skipping of exon 6. The mutation in cop1-8 was G- >A in the final nucleotide of intron 10 abolishing the conserved G within the 3' splice site consensus and leading to skipping of exon 11. The splicing patterns surrounding exons 6 and 11 of COP1 in these three mutant lines of Arabidopsis provide evidence for exon definition mechanisms operating in plant splicing. PMID- 9744101 TI - A naturally occurring functional allele of the rice waxy locus has a GT to TT mutation at the 5' splice site of the first intron. AB - In cultivated rice two wild-type alleles, Wxa and Wxb, predominate at the waxy locus, which encodes granule-bound starch synthase. The activity of Wxa is 10 fold higher than that of Wxb at the level of both protein and mRNA. Wxb has a +1G to T mutation at the 5' splice site of the first intron. Sequence analysis of Wxb transcripts revealed that splicing occurs at the mutant AG/UU site and at two cryptic sites: the first is A/GUU, one base upstream of the original site and the second is AG/GU found approximately 100 bases upstream of the mutant splice site. We introduced single base mutations to the 5' splice sites of both Wxa and Wxb, fused with the gus reporter gene and introduced them into rice protoplasts. Analysis of GUS activities and transcripts indicated that a G to T mutation in Wxa reduced GUS activity and the level of spliced RNA. Conversely, a T to G mutation of Wxb restored GUS activity and the level of spliced RNA to that of wild-type Wxa. These results demonstrated that the low level expression of Wxb results from a single base mutation at the 5' splice site of the first intron. It is of interest that the Wxb allele of rice carrying the G to T mutation of intron 1 has been conserved in the history of rice cultivation because there is a low amylose content of the seed caused by this mutation. PMID- 9744102 TI - Phytepsin, a barley vacuolar aspartic proteinase, is highly expressed during autolysis of developing tracheary elements and sieve cells. AB - Vacuolarisation, formation of autophagocytotic vacuoles and tonoplast disruption have been reported in plant cells undergoing developmentally regulated programmed cell death (PCD), but little is known about the vacuolar proteins involved. In HeLa cells, cathepsin D, a lysosomal aspartic proteinase has been shown to mediate PCD. Based on immunohistochemical staining of barley roots, we show here that the previously well characterised barley vacuolar aspartic proteinase (phytepsin), a plant homologue to cathepsin D, is highly expressed both during formation of tracheary elements and during partial autolysis of sieve cells. In serial transverse sections of the vascular cylinder, starting from the root tip, phytepsin is expressed in root cap cells, in the tracheary elements of early and late metaxylem, and in the sieve cells of the protophloem and metaphloem. Aleurain, a barley vacuolar cysteine proteinase, is expressed similarly in root cap cells but differently in the tracheary elements of protoxylem and early metaxylem. This is the first evidence that a vacuolar aspartic proteinase, in analogy to cathepsin D in animals, may play a role in the active autolysis of plant cells. PMID- 9744103 TI - Rapid parapatric speciation on holey adaptive landscapes. AB - A classical view of speciation is that reproductive isolation arises as a by product of genetic divergence. Here, individual-based simulations are used to evaluate whether the mechanisms implied by this view may result in rapid speciation if the only source of genetic divergence are mutation and random genetic drift. Distinctive features of the simulations are the consideration of the complete process of speciation (from initiation until completion), and of a large number of loci, which was only one order of magnitude smaller than that of bacteria. It is demonstrated that rapid speciation on the time-scale of hundreds of generations is plausible without the need for extreme founder events, complete geographic isolation, the existence of distinct adaptive peaks or selection for local adaptation. The plausibility of speciation is enhanced by population subdivision. Simultaneous emergence of more than two new species from a subdivided population is highly probable. Numerical examples relevant to the theory of centrifugal speciation and to the conjectures about the fate of 'ring species' and 'sexual continuums' are presented. PMID- 9744104 TI - Spatial asynchrony and periodic travelling waves in cyclic populations of field voles. AB - We demonstrate evidence for the presence of travelling waves in a cyclic population of field voles in northern Britain by fitting simple, empirical models to spatially referenced time series data. Population cycles were broadly synchronous at all sites, but use of Mantel correlations suggested a strong spatial pattern along one axis at a projection line 72 degrees from North. We then fitted a generalized additive model to log population density assuming a fixed-form travelling wave in one spatial dimension for which the density at each site was offset in time by a constant amount from a standard density-time curve. We assumed that the magnitude of this offset would be proportional to the spatial separation between any given site and the centroid of the sampling sites, where separation is the distance between sites in a fixed direction. After fitting this model, we estimated that the wave moved at an average speed of 19 km yr-1, heading from West to East at an angle of 78 degrees from North. Nomadic avian predators which could synchronize populations over large areas are scarce and the travelling wave may be caused by density-dependent dispersal by field voles and/or predation by weasels, both of which act at a suitably small spatial scale. PMID- 9744105 TI - Fluctuating asymmetry and human male life-history traits in rural Belize. AB - Fluctuating asymmetry (FA), used as a measure of phenotypic quality, has proven to be a useful predictor of human life-history variation, but nothing is known about its effects in humans living in higher fecundity and mortality conditions, typical before industrialization and the demographic transition. In this research, I analyse data on male life histories for a relatively isolated population in rural Belize. Some of the 56 subjects practise subsistence-level slash-and-burn farming, and others are involved in the cash economy. Fecundity levels are quite high in this population, with men over the age of 40 averaging over eight children. Low FA successfully predicted lower morbidity and more offspring fathered, and was marginally associated with a lower age at first reproduction and more lifetime sex partners. These results indicate that FA may be important in predicting human performance in fecundity and morbidity in predemographic transition conditions. PMID- 9744106 TI - The curse of the pharaoh hypothesis. AB - The 'curse of the pharaoh' has been used as a metaphor for the hypothesis that higher parasite propagule survival selects for higher virulence. Indeed, the mysterious death of Lord Carnavon after entering the tomb of the Egyptian pharaoh Tutankhamen could potentially be explained by an infection with a highly virulent and very long-lived pathogen. In this paper, I investigate whether parasite virulence increases with high propagule survival. In this respect, I derive an analytic expression of the evolutionarily stable level of parasite virulence as a function of propagule survival rate when the host-parasite system has reached a stable ecological equilibrium. This result shows that, if multiple infection occurs, higher propagule survival generally increases parasite virulence. This effect is enhanced when parasite dispersal coevolves with parasite virulence. In a more general perspective, the model shows the importance of taking into account the combination of direct and indirect effects (which I call inclusive effects) of higher transmission ability on the evolution of parasite virulence. The recognition of these effects has several practical implications for virulence management. PMID- 9744107 TI - Trade-off associated with selection for increased ability to resist parasitoid attack in Drosophila melanogaster. AB - Costs of resistance are widely assumed to be important in the evolution of parasite and pathogen defence in animals, but they have been demonstrated experimentally on very few occasions. Endoparasitoids are insects whose larvae develop inside the bodies of other insects where they defend themselves from attack by their hosts' immune systems (especially cellular encapsulation). Working with Drosophila melanogaster and its endoparasitoid Leptopilina boulardi, we selected for increased resistance in four replicate populations of flies. The percentage of flies surviving attack increased from about 0.5% to between 40% and 50% in five generations, revealing substantial additive genetic variation in resistance in the field population from which our culture was established. In comparison with four control lines, flies from selected lines suffered from lower larval survival under conditions of moderate to severe intraspecific competition. PMID- 9744108 TI - Molecular and functional characterization of a recombinant protein of Trichuris trichiura. AB - The pore-forming protein of the human whipworm, Trichuris trichiura, has been postulated to facilitate invasion of the host gut and enable the parasite to maintain its syncytial environment. The data presented here describe the first, to our knowledge, molecular characterization of a pore-forming protein in any helminth and provide a unique demonstration of the functional interaction between a parasite antigen and host molecules. Immunological screening of a T. trichiura cDNA library with T. trichiura infection sera identified a clone of 1.4 kB, the cDNA consisting of 1495 base pairs encoding a protein of 50 kDa. The sequence has a highly repetitive nature containing nine four-disulphide-bonded core domains. Structural prediction analyses reveals an amphipathic nature. TT50 induced pore formation in bilayers in a manner identical to that of the native protein. IgG antibody isolated from T. trichiura infection serum was observed to abolish channel activity. PMID- 9744109 TI - Stereoscopic and contrast-defined motion in human vision. AB - There is considerable evidence for the existence of a specialized mechanism in human vision for detecting moving contrast modulations and some evidence for a mechanism for detecting moving stereoscopic depth modulations. It is unclear whether a single second-order motion mechanism detects both types of stimulus or whether they are detected separately. We show that sensitivity to stereo-defined motion resembles that to contrast-defined motion in two important ways. First, when a missing-fundamental disparity waveform is moved in steps of 0.25 cycles, its perceived direction tends to reverse. This is a property of both luminance defined and contrast-defined motion and is consistent with independent detection of motion at different spatial scales. Second, thresholds for detecting the direction of a smoothly drifting sinusoidal disparity modulation are much higher than those for detecting its orientation. This is a property of contrast modulated gratings but not luminance-modulated gratings, for which the two thresholds are normally identical. The results suggest that stereo-defined and contrast-defined motion stimuli are detected either by a common mechanism or by separate mechanisms sharing a common principle of operation. PMID- 9744110 TI - The asynchrony of consciousness. AB - We present below a simple hypothesis on what we believe is a characteristic of visual consciousness. It is derived from facts about the visual brain revealed in the past quarter of a century, but it relies most especially on psychophysical evidence which shows that different attributes of the visual scene are consciously perceived at different times. This temporal asynchrony in visual perception reveals, we believe, a plurality of visual consciousnesses that are asynchronous with respect to each other, reflecting the modular organization of the visual brain. We further hypothesize that when two attributes (e.g. colour and motion) are presented simultaneously, the activity of cells in a given processing system is sufficient to create a conscious experience of the corresponding attribute (e.g. colour), without the necessity for interaction with the activities of cells in other processing systems (e.g. motion). Thus, any binding of the activity of cells in different systems should be more properly thought of as a binding of the conscious experiences generated in each system. PMID- 9744111 TI - Industrial platforms--a unique feature of the European Commission's biotechnology R&D programme. AB - The European Commission's research, technological development and demonstration programmes aim to strengthen European research and technological development, and to increase the competitiveness of European industries. The creation and development of Industrial Platforms play an important role in these processes by improving the transition from research to commercial application. Industrial Platforms are technology-based industrial groupings established by industry with the aims of enabling the exploitation or dissemination of research results, encouraging academic-industrial collaborations and providing their members with a means of voicing their opinion on present and future research policies. PMID- 9744112 TI - Systematic functional analysis of the yeast genome. AB - The genome sequence of the yeast Saccharomyces cerevisiae has provided the first complete inventory of the working parts of a eukaryotic cell. The challenge is now to discover what each of the gene products does and how they interact in a living yeast cell. Systematic and comprehensive approaches to the elucidation of yeast gene function are discussed and the prospects for the functional genomics of eukaryotic organisms evaluated. PMID- 9744113 TI - Microbial denitrogenation of fossil fuels. AB - The microbial degradation of nitrogen compounds from fossil fuels is important because of the contribution these contaminants make to the formation of nitrogen oxides (NOx) and hence to air pollution and acid rain. They also contribute to catalyst poisoning during the refining of crude oil, thus reducing process yields. We review the current status of microbial degradation of aromatic nitrogen compounds and discuss the potential of microbial processes to alleviate these problems. PMID- 9744115 TI - Thirsting for attention. PMID- 9744114 TI - Microbial lipases form versatile tools for biotechnology. AB - Lipases are secreted into the culture medium by many bacteria and fungi. They catalyse not only the hydrolysis but also the synthesis of long-chain acylglycerols. Important uses in biotechnology include their addition to detergents, the manufacture of food ingredients, pitch control in the pulp and paper industry, and biocatalysis of stereoselective transformations. This makes them the most widely used class of enzymes in organic chemistry. Immobilization in hydrophobic sol-gel matrices and in vitro evolution are promising novel approaches to increasing the stability or enantioselectivity, respectively, of lipases. PMID- 9744116 TI - Conversation with David Hawks. PMID- 9744117 TI - The quality of alcohol treatment research: an examination of influential controlled trials and development of a quality rating system. AB - BACKGROUND: The importance of evidence-based practice has stimulated interest in the methodology of clinical trials. Various weaknesses of evaluation research in the alcohol field have been indicated previously. This study set out to develop a comprehensive system for the assessment of the methodological quality of outcome research for treatment of alcohol misuse and to apply the system to well-known trials in the area. METHODOLOGY: A sample of the most highly cited controlled trials of interventions for alcohol misuse was selected using the Science Citation Index. Thirty methodological criteria were formulated and a scoring system devised. Two raters applied this system to the sample of trials. Reliability testing was performed and used to refine the criteria. RESULTS: Inter rater reliability of the overall quality score was initially 0.85 and 0.92 after review and re-rating. Internal consistency was also high (0.87). Quality score correlated with year of publication. Certain areas of methodology were poorly addressed in the sample, including specification of main outcomes, documentation of recruitment and selection procedures, testing of blinding, analysis of withdrawals and reporting of results. DISCUSSION: The methodology of this sample of trials was frequently deficient in ways which might bias results or compromise generalizability. It is recommended that the system of quality assessment described here is used to evaluate existing research and to inform the design of future studies. PMID- 9744118 TI - Effects of "binge" use of intravenous cocaine in methadone-maintained individuals. AB - AIMS: To examine the effects of heavy cocaine use followed by abstinence. DESIGN: During self-administration sessions, which occurred in the afternoon and again in the evening, participants could after self-administer up to six doses of intravenous cocaine (32 mg/70 kg) at 14-minute intervals. All volunteers participated in a 2-day and a 3-day series of self-administration sessions (cocaine binges). No cocaine was available for at least 2 days following each cocaine binge. PARTICIPANTS: Five adult methadone-maintained cocaine abusers. SETTING: Inpatient clinical research unit. MEASUREMENTS: Cardiovascular and subjective effects. FINDINGS: The first cocaine dose in a session produced substantial subjective and cardiovascular effects and, with the exception of blood pressure and ratings of "irritable", administering up to five more cocaine doses did not increase these effects. Additional cocaine doses, however, did increase ratings of "bad drugs effect" and decrease ratings of "I want cocaine". Maximal subjective and cardiovascular effects of cocaine did not vary across sessions on the same day or between days. Ratings of "depressed" and "miserable" were increased the first day after each period of cocaine use, and presentation of cocaine-related stimuli increased ratings of "I want cocaine" and "I want heroin" during cocaine abstinence. CONCLUSION: Acute tolerance developed to many of the effects of cocaine during a cocaine binge in methadone-maintained individuals, next-day residual changes in mood were subtle, and no prolonged effects of a cocaine binge were observed. PMID- 9744119 TI - Long-term cannabis use: characteristics of users in an Australian rural area. AB - AIM: To investigate the characteristics and patterns of cannabis and other drug use among long-term cannabis users in an Australian rural area. DESIGN: Cross sectional survey of a "snowball" sample of long-term cannabis users. SETTING: The North Coast of New South Wales is an area with high levels of cannabis cultivation and use, and many long-term users. PARTICIPANTS: The study involved 268 long-term cannabis users who had regularly used cannabis for at least 10 years. MEASUREMENTS: A structured interview schedule obtained information on: demographics, social circumstances, patterns of cannabis and other drug use, contexts of use, perceptions about cannabis and legal involvement. FINDINGS: The mean age of the sample was 36 years and 59% were made. The median length of regular cannabis use was 19 years. Most (94%) used two or more times a week and 60% used daily, with a median of two joints per day. Two-thirds (67%) used cannabis in social settings and two-thirds grew cannabis for their own use. The most common reasons for using cannabis were for relaxation or relief of tension (61%) and enjoyment or to feel good (27%). The most commonly reported negative effects were feelings of anxiety, paranoia, or depression (21%), tiredness, lack of motivation and low energy (21%) and effects of smoke on the respiratory system (18%). The majority drank alcohol (79%) and over one-third were drinking at hazardous levels. Most were current (64%) or ex-tobacco smokers (24%). One quarter (25%) had been charged with possession of cannabis, 11% for cultivation and 6% for supply, with non-drug offences low (8%or less). Overall, three quarters (72%) believed that the benefits of cannabis use outweighed the risks, 21% felt there was an even balance, and 7% said cannabis had done them more harm than good. CONCLUSIONS: Among long-term cannabis users in this Australian rural area, cannabis use was an integral part of everyday life and it was primarily used in social situations for the same reasons that alcohol use is used in the wider community. PMID- 9744120 TI - Detection of drug use in a methadone maintenance clinic: sweat patches versus urine testing. AB - AIMS: To evaluate the novel use of sweat patches in outpatients attending a methadone maintenance clinic. DESIGN: Assessment of inter-patch reliability and validity of patch results compared to urine tests at the start and end of the patch period. Semi-structured questionnaire on patients' opinion of the patches. Randomized cross-over trial comparing illicit drug use during the week that the patch was worn with a control period. SETTING: Methadone maintenance outpatient clinic in a deprived urban area. PARTICIPANTS: Forty-eight patients with a diagnosis of opiate addiction prescribed methadone for a median of 5 years. MEASUREMENTS: Analysis of urine and patch tests by standard methods for methadone, opiates, morphine (heroin metabolite) and benzoylecgonine (cocaine metabolite). FINDINGS: There was good inter-patch reliability between arm and side patches for methadone (all positive), opiates (k = 0.8) and morphine (k = 1.0) but only moderate agreement for benzoylecgonine (k = 0.49). There was good agreement between the sweat patches and urine tests for methadone (all positive), opiates (k = 1.00) and morphine (k = 0.8), but again only moderate agreement for benzoylecgonine (k = 0.50). The patches were well tolerated and the main side effect was minor irritation. The majority of men preferred a urine test while more women preferred a sweat patch. There was no evidence of reduced use of illicit drugs the period that the patch was worn. CONCLUSION: Sweat patches are reliable and give valid results for patients on maintenance methadone. There are few side effects and it is practical to use them in busy outpatient clinics. They are preferred by some patients but there was no evidence that they altered behaviour. PMID- 9744121 TI - A comparison of the prevalence of HIV infection and injecting risk behaviour in urban and rural samples in Scotland. AB - AIMS: To compare the prevalence of HIV infection and injecting risk behaviour among injecting drug users from urban and rural areas. DESIGNS: Two samples of injecting drug users were recruited using a multi-site sampling strategy. Respondents were first interviewed by trained interviewers then specimens of saliva were collected for anonymous testing for antibodies to HIV. SETTING: Respondents were recruited from drug treatment services and street sites. PARTICIPANTS: Respondents were eligible for inclusion in the study if they had "ever" injected a drug and were currently using drugs. MEASUREMENTS: Measurements taken included self-reported patterns of drug use and injecting risk behaviour. Specimens of saliva were tested for the presence of HIV infection using GACELISA with reactive specimens confirmed by Western blot analysis. FINDINGS: Our data indicate that there are two separate populations which are geographically discrete but broadly similar in profile and current injecting risk behaviour. The prevalence of HIV infection among IDUs from Dundee city was found to be 26.8% (95% CI, 20.2%-33.0%) compared with 3.7% (95% CI, 0.13%-15.8%) for IDUs from rural Tayside. This marked difference in prevalence of HIV infection we attribute to a high level of injecting risk among urban IDUs between 1980 and 1984, limited migration from the urban epicentre of infection and a reluctance among rural IDUs to share with IDUs outside their immediate social and kinship networks. CONCLUSIONS: Although current levels of injecting risk behaviour are similar in our urban and rural samples, rural IDUs may be less likely to contract HIV from their fellow injectors because of the lower prevalence of HIV infection and more closed sharing networks within the rural population. The implications of this for the development and expansion of drug services are considered. PMID- 9744122 TI - Exploring the nature of the relationship between child sexual abuse and substance use among women. AB - AIMS: This study investigated whether child abuse (CSA) was associated with earlier substance use and greater severity of substance dependence and what aspects of CSA might predict substance abuse. DESIGN: The study compared (a) drug and alcohol treatment clients with and without a history of CSA and (b) CSA survivors outside drug and alcohol treatment who did or did not have current substance abuse. SETTINGS: Semi-structured interviews took place at participants' homes, treatment agencies or the research centre. PARTICIPANTS: Volunteer participants included 100 women recruited from drug and alcohol treatment programmes and 80 CSA survivors recruited through CSA counseling services and medial advertising. MEASUREMENTS: The results focus on data from the Opiate Treatment Index, Severity of Alcohol Dependence Questionnaire, Substance Dependence Scale, Self-Esteem Inventory and self-reported histories of CSA. FINDINGS: There were no differences between CSA survivors and other drug and alcohol treatment clients in their severity of dependence. Women with a history of CSA more frequently identified stimulants as their main problem drug and reported an earlier age of first intoxication and earlier use of inhalants. Among abused CSA survivors outside drug and alcohol treatment, women with current substance abuse had typically been abused during adolescence by someone outside the family, whereas those without current substance abuse were typically abused by family members before adolescence. CONCLUSIONS: The results suggest that adolescence is a crucial time for the influence of CSA experiences on substance abuse. PMID- 9744123 TI - Effect of smoking cessation counseling on recovery from alcoholism: findings from a randomized community intervention trial. AB - AIMS: To assess the effects of a smoking cessation program for recovering alcoholics on use of alcohol, tobacco and illicit drugs after discharge from residential treatment. DESIGN AND SETTING: A randomized community intervention trial design was employed in which 12 residential drug treatment centers in Iowa, Kansas and Nebraska were matched and then randomly assigned to the intervention or control condition. PARTICIPANTS: Approximately 50 adult residents (inpatients) from each site were followed for 12 months after treatment discharge. INTERVENTION: Participating residents in the six intervention centers received a 4-part, individually tailored, smoking cessation program while those in the six control sites received usual care. FINDINGS: Both moderate and heavy drinking rates were reduced in the intervention group. Intervention site participants were significantly more likely than controls to report alcohol abstinence at both the 6-month (OR = 1.59, 95%CI: 1.09-2.35) and 12-month assessment (OR = 1.84, 95%CI: 1.28-2.92). Illicit drug use rates were comparable. Effect of the intervention on tobacco quit rates was not statistically significant. CONCLUSIONS: Counseling alcoholics in treatment to quit smoking does not jeopardize the alcohol recovery process. However, low-intensity tobacco interventions are unlikely to yield high tobacco quit rates. PMID- 9744124 TI - Psychiatric co-morbidity, suicidal behaviour and suicidal ideation in alcoholics seeking treatment. AB - AIMS: To estimate the impact of co-morbid disorders for suicidal ideas in alcohol dependent subjects seeking treatment. DESIGN: Life-time psychiatric co-morbidity and previous suicidal behaviours were assessed retrospectively after detoxification (t1). In addition, suicidal behaviours were assessed 12 months after discharge (t2). SETTING: An inpatient detoxification treatment unit. PARTICIPANTS: Two hundred and fifty dependent inpatients were studied after detoxification. One hundred and forty-nine of them participated in the follow-up face-to-face interviews. MEASUREMENTS: Using two extended standardized interviews (CIDI and IPDE) psychiatric co-morbidity (DSM-III-R, Axes I and II) was assessed at t1; suicide attempts were reported at t1 and t2, and suicidal ideas were assessed at t2. FINDINGS: A history of suicide attempts was reported by 29.2% at t1, and suicidal ideas by 14.1% and suicide attempts by 5.4% at the follow-up (t2). One female patient committed suicide within 6 months of discharge from hospital. The following co-morbidity patterns were associated with the greatest risk for suicidal ideas. Anxiety and depressive disorders, Axes I and 11 disorders, and a history of suicide attempt (for suicidal ideas at (t2). CONCLUSION. Our results underline the importance of psychiatric co-morbidity for the suicidal risk in alcohol-dependent patients, while alcoholism itself appears to be only a moderate risk factor. PMID- 9744125 TI - Impulsivity, personality disorders and pathological gambling severity. AB - AIMS: The present study seeks to replicate and expand findings from earlier studies that pathological gamblers manifest elevated traits of impulsivity. Secondly, the study aims to elucidate the relationship between impulsivity, indices reflecting severity of pathological gambling and other measures of psychopathology and personality dysfunction. DESIGN: Case series. PARTICIPANTS: Eighty-two consecutive gamblers, seeking treatment for problem gambling. SETTING: Impulse Disorders Research Unit, School of Psychiatry, University of New South Wales, Australia. MEASUREMENTS: Semi-structured interview designed to obtain demographic information and gambling history, the South Oaks gambling Screen, the Eysenck Impulsivity Scale, the Personality Disorder Questionnaire-Revised, the Beck Depression Inventory and the Beck Anxiety Inventory. FINDINGS: Results show elevated traits of impulsivity among clinic samples of pathological gamblers compared to normative data and show that impulsivity is related to the severity of gambling behaviour as measured by the South Oaks Gambling Questionnaire. A principal component analysis of the Eysenck Impulsivity Scale and the personality Disorder Questionnaire-Revised, demonstrated that the concept of the "antisocial impulivist" identified to Blaszczynski, Steel & McConaghy (1997) is not only characterized by impulsivity and antisocial personality disorder but also by high loadings from other cluster B and cluster C personality disorders. CONCLUSION. This research supports the role of the construct of impulsivity in mediating the severity of gambling behaviour and associated behavioural and psychological disturbance among pathological gamblers presenting for treatment. Impulsivity is best understood as part of a general personality disorder structure characterized primarily by DSM-III-R Axis II cluster B and some cluster C personality disorder. PMID- 9744126 TI - Zopiclone, a current drug of misuse. PMID- 9744127 TI - Measuring alcohol consumption: limitations and prospects for improvement. PMID- 9744129 TI - Researching the spiritual dimensions of alcohol and other drug problems. AB - Although religions have been far from silent on the use of psychoactive drugs, and spirituality has long been emphasized as an important factor in recovery from addiction, surprisingly little research has explored the relationships between these two phenomena. Current findings indicate that spiritual/religious involvement may be an important protective factor against alcohol/drug abuse. Individuals currently suffering from these problems are found to have a low level of religious involvement, and spiritual (re)engagement appears to be correlated with recovery. Reasons are explored for the lack of studies testing spiritual hypotheses, and promising avenues for future research are discussed. Comprehensive addictions research should include not only biomedical, psychological and socio-cultural factors but spiritual aspects of the individual as well. PMID- 9744130 TI - The economic costs of alcohol, tobacco and illicit drugs in Canada, 1992. AB - AIMS, DESIGN AND SETTING: The economic costs of alcohol, tobacco and illicit drugs in Canadian society in 1992 are estimated utilizing a cost-of-illness framework and recently developed international guidelines. MEASUREMENTS: For causes of disease or death (using ICD-9 categories), pooled relative risk estimates from meta-analyses are combined with prevalence data by age, gender and province to derive the proportion attributable to alcohol, tobacco and/or illicit drugs. The resulting estimates of attributable deaths and hospitalizations are used to calculate associated health care, law enforcement, productivity and other costs. The results are compared wit other studies, and sensitivity analyses are conducted on alternative measures of alcohol consumption, alternative discount rates for productivity costs and the use of diagnostic-specific hospitalization costs. FINDINGS: The misuse of alcohol, tobacco and illicit drugs cost more than $18.4 billion in Canada in 1992, representing $649 per capita or 2.7% of GDP. Alcohol accounts for approximately $7.52 billion in costs, including $4.14 billion for lost productivity, $1.36 billion for law enforcement and $1.30 billion in direct health care costs. Tobacco accounts for approximately $9.56 billion in costs, including $6.82 billion for lost productivity and $2.68 billion for direct health costs. The economic of illicit drugs are estimated at $1.4 billion. CONCLUSIONS: Substance abuse exacts a considerable toll to Canadian society in terms of illness, injury, death and economic costs. PMID- 9744131 TI - A randomized controlled trial of a "buddy" systems to improve success at giving up smoking in general practice. AB - AIMS: To assess the effect on abstinence rates of pairing up smokers attending a general practice smokers, clinic to provide mutual support between clinic sessions. DESIGN: Randomized controlled trial comparing a "buddy" condition with a "solo condition" in which smokers received the e same treatment but were not paired up. SETTING: A general practice smokers' clinic in London. PARTICIPANTS: One hundred and seventy-two smokers recruited by mailshot. INTERVENTION. Smokers attended a nurse-led smokers clinic 1 week prior to their quit date, on the quite date, 1 week later and 3 weeks after that. Smokers in the buddy condition were paired with another smoker trying to give up at the same time to provide mutual support between clinic sessions. MEASUREMENT: The main outcome measure was the percentage of smokers still abstinent from cigarettes at end of treatment (weeks from quite date), verified by expired air carbon monoxide concentration. FINDINGS: The percentage of smokers still abstinent at the end of treatment was significantly higher in the buddy condition than the solo condition (27% vs. 12%). CONCLUSIONS: A buddy system can provide an effective element of a smoking cessation intervention at minimal cost. Further research is needed to establish the long-term efficacy of this approach and examine the effectiveness of incorporating social support into other types of smoking cessation programmes. PMID- 9744132 TI - General practitioners' role in preventive medicine: scenario analysis using smoking as a case study. AB - AIM: It is the purpose of this paper to develop a model which may be used in conjunction with scenario analysis to evaluate strategies which are available to assist the general practitioner (GP) in reducing smoking behaviour among their patients. DESIGN: The scenario analysis uses a four-step procedure which involves identifying opportunities for detection, intervention and efficacy, and assigning probabilities to outcome to enable a range of prevention strategies to be examined in both isolation and in combination. SETTING AND PARTICIPANTS: This study deals specifically with Australian general practice and the model is derived by using information for a smoker visiting their GP within a 6-month period together with empirical evidence on the rates of detection, intervention and efficacy. MEASUREMENTS: The outcome measures, which are evaluated in terms of marginal effectiveness, include the number of smoking patients detected, the number of smoking patients offered an intervention, the number of smoker patients who quit as a result of the intervention and the additional years of life saved due to an intervention. FINDINGS. The most significant indicator for reducing smoking rates among patients is improving the efficacy of interventions. The results also suggest that although improvements in the rate of GP detection of patients' smoking status have a potentially greater effect on quit rates than increasing intervention levels, increasing both detection and intervention levels had a greater effect than each strategy alone. DISCUSSION: : General practitioners have an important role to play in preventive medicine. The knowledge, skill and attitude of practitioners toward smoking are significant, and they can be the prime motivators is persuading their patients to stop smoking. Detection, intervention and efficacious strategies are all key elements in achieving this result. PMID- 9744133 TI - Life-time prevalence and risk factors of tobacco/nicotine dependence in male ever smokers in Japan. AB - To estimate the life-time prevalence rate of tobacco/nicotine dependence and demographic variables and smoking habits associated with the disorder in male ever-smokers in Japan. DESIGN: A cross-sectional community-based interview study. SETTING: Takayama city, Gifu Prefecture, Japan. PARTICIPANTS: A total of 170 male ever-smokers aged 35 years or older selected randomly from a community in Japan were interviewed. The response rate was 85%. MEASUREMENTS: The WHO Composite International Diagnostic Interview (CIDI) was used to make diagnoses of tobacco/nicotine dependence according t ICD-10, DSM-III-R and DSM-IV. The Fagerstrom Tolerance Questionnaire (FTQ) was also administered and those who had a FTQ score of 7 or above were identified. FINDINGS: The life-time prevalence rates of tobacco/nicotine dependence in male ever-smokers were 42%, 26% and 32% according to ICD-10, DSM-III-R and DSM-IV criteria, respectively; 19% had a FTQ score of 7 or above. The ICD-10 diagnosis was significantly and negatively associated with quitting smoking (p < 0.05). Multiple logistic regression analyses indicated that number of cigarettes per day when they smoked the most was significantly associated with higher life-time risks of the disorder according to DSM-III-R, DSM-IV and Fagerstrom's classification (p < 0.05). The length of cigarette smoked was associated with higher life-time risks of ICD-10 and DSM-IV diagnoses, and years of smoking were associated with higher life-time risks of ICD-10, DSM-III-R and DSM-IV diagnoses (p < 0.05). Younger birth cohorts had higher cumulative rates of the disorder according to DSM-IV (p for trend < 0.05). CONCLUSIONS: Life-time prevalence rates of tobacco/nicotine dependence according to ICD-10, DSM-III-R and DSM-IV in male ever-smokers in Japan were within the range of rates reported in previous US studies; rates of FTQ score of 7 or above were lower. Fagerstrom scores and diagnostic criteria appear to reflect different aspects of dependence. PMID- 9744134 TI - Assessing the relationship between maternal opiate use and neonatal mortality. AB - We undertook a number of meta-analyses to estimate more precisely the relationship between neonatal mortality and use of opiates in three groups of women. First, women who continued to use illicit heroin throughout pregnancy; secondly, women stabilized on methadone at the time of conception or shortly after and thirdly, women who use heroin well into pregnancy with late entry into methadone treatment, or who continued to use illicit heroin during pregnancy while receiving methadone. FINDINGS: The pooled estimates of the relative risks of neonatal mortality for separate heroin and methadone use were both near unity: 1.47 (95% CI 0.88-2.33) and 1.75 (95% CI 0.60-4.59), respectively. The result for heroin may be due to the inclusion in the meta-analysis of a particularly large study, which, unlike the two other smaller studies included found a relative risk near unity. When this study was excluded from the meta-analysis the pooled estimate of the relative risk of neonatal mortality for heroin use was 3.27 (95% CI 0.95-9.60). In contrast to the results for use of methadone only, the pooled relative risk associated with heroin and methadone use was 6.37 (95% CI 2.57 14.68). CONCLUSIONS: The increased relative risk for neonatal mortality associated with women using heroin and methadone during pregnancy, compared to those stabilized on methadone, is probably due to the chaotic and high-risk life style associated with illicit heroin use and not solely to the use of heroin and methadone per se. It is recommended tht women who use heroin well into pregnancy with late entry into methadone treatment, or who continue to use illicit heroin during pregnancy while receiving methadone, receive special attention over and above that provided to women stabilized on methadone. PMID- 9744135 TI - Therapist/patient race and sex matching: treatment retention and 9-month follow up outcome. AB - AIMS: The current study was conducted to (a) replicate previous findings regarding the effect of patient/therapist race and sex-matching as this relates to the early dropout rate of substance abusers, and (b) to extend previous work by examining the impact of such matching on treatment retention and 9-month outcome. DESIGN: Patient and therapist characteristics were crossed in a 2 x 2 factorial design. Matching effects were then tested using retrospective data. PARTICIPANTS: Participants were 967 African-American cocaine-dependent people. SETTING: The study site was a university sponsored outpatient treatment facility located in an economically depressed section of a large Northeastern US city. MEASUREMENTS: Follow-up data were drawn from the Addiction Severity Index, the Risk for AIDS Behavior Inventory, and a questionnaire measuring post-discharge need for treatment, employment and education. Retention was defined as the number of days inclusive between the last date of service and the date of admission. FINDINGS: No significant differences in early dropout rates were found after initial contact with 10 different therapists. Matching therapist and patients with respect to gender and race did not decrease the premature dropout rate, but partial support for gender matching was noted. CONCLUSION: Matching therapist and substance abusing patients on gender and race may not be essential to improving retention and outcome. PMID- 9744136 TI - Treatment level progress and time spent in treatment in the prediction of outcomes following drug-free therapeutic community treatment. AB - Previous research has suggested that time spent in treatment is the most important predictor of therapeutic community treatment outcomes. Although level divisions or treatment stages represent the basic structure for segmenting responsibility and authority within traditional therapeutic communities, the relationship of this treatment component to treatment outcome has not previously been investigated. AIM: To test the hypothesis that higher treatment level attainment, more time spent in treatment and additional time spent within a treatment level has a linear association with improvements at outcome. DESIGN: Retrospective quasi-experimental. PARTICIPANTS: Four hundred and twenty-seven ex residents, stratified according to their highest level of treatment in the Melbourne Odyssey House therapeutic community between 1984 and 1988 were targeted for follow-up and 60% were successfully located and interviewed an average of 5.6 years after their first Odyssey entry. Analysis of official records of methadone treatment, convictions and incarcerations demonstrated no post-treatment differences comparing those interviewed with those not interviewed. MEASUREMENTS: Drug use, criminal involvement and employment. FINDINGS. Although both level attainment and time spent in treatment had a linear relationship to improved outcomes, level attainment was a better predictor of outcomes at treatment exit. An unexpected finding was that those who had spent the median time or longer in particular levels demonstrated worse outcomes on official conviction records and on self-reports of employment compared to those remaining for less than the median time. CONCLUSION: The results suggest that it may be level progress rather than simply time spent in treatment that best explains improved functioning following exit from the therapeutic community. PMID- 9744137 TI - DSM-IV alcohol disorders in a general population sample of adolescents and young adults. AB - AIMS/DESIGNS: As part of the Early Developmental Stages of Psychopathology (EDSP) study, results from the baseline cross-sectional assessment of DSM-IV alcohol disorders are presented for a sample of 14-24-year-olds residents in Munich, Germany (N = 3021; 71% response rate). FINDINGS: Life-time prevalence of DSM-IV alcohol abuse (men: 15.1%; women; 4.5%) was found to be considerably more prevalent than dependence (men: 10.0%; women 2.5%) with few cases among respondents younger than 16 years of age; 12-month prevalence of abuse was 8.4% among men and 2.7% among women and of dependence was 7.3% among men and 2.2% among women. Results show that peak incidence of alcohol disorders occurs at 16 17 years of age and that early initiation into alcohol use is associated with an increasing odds of disorder onset, especially for dependence among women. Exploratory analysis of retrospectively assessed diagnostic stability show: a temporal progression to abuse and then dependence, that nearly half of past abuse diagnoses are in remission, abuse remission is more common than progression to dependence, and dependence is highly persistent, especially among women. CONCLUSIONS: Alcohol disorders are frequent in adolescent and young adults being characterized by transient abuse and less prevalent but persistent dependence syndromes. The relatively high prevalence of dependence diagnoses in this young population wit few years of alcohol use is discussed with regard to the clinical validity of DSM-IV criteria in adolescents and young adults. PMID- 9744138 TI - Drinking patterns, psychosocial characteristics and alcohol consequences. AB - AIMS: The purpose of this study was to investigate (a) the potential mediating role of alcohol consumption in the relationship between psychosocial variables and adverse consequences of drinking, and (b) the possible moderating role of psychosocial variables in the relationship between alcohol consumption and adverse consequences. DESIGN: Multi-stage sampling procedures were used to select households for a general population survey. Interviews were conducted with one member in each of 35, 479 households and self-administered questionnaires were left for all residents aged 12 years and older (N = 49 164, response rate: 77.2%). SETTING: Data were collected as part of the Ontario (Canada) Health Survey. PARTICIPANTS: The present study included 33, 568 current drinkers aged 18 or older. MEASUREMENTS: Analyses included the following variables: alcohol consumption, adverse consequences related to drinking, sex, age, marital status, self-rated health, perceived stress, income, education, employment status and family dysfunction. FINDINGS: Alcohol consumption appeared to mediate the relationship of adverse consequences with age, sex, marital status, education, income and employment, but not with health, stress or family dysfunction which were independently associated with adverse consequences. The relationship between alcohol consumption and adverse consequences was moderated by age, marital status, education and employment. CONCLUSIONS: These results help to clarify the interrelationship of alcohol consumption and psychosocial characteristics in experiencing adverse consequences related to drinking. PMID- 9744139 TI - A comparison of two alcohol craving questionnaires. AB - AIM: To compare two multi-dimensional questionnaires to measure cravings and urges for alcohol: the Alcohol Craving Questionnaire (ACQ: Singleton, Henningfield and Tiffany, 1994a) and the Desires for Alcohol Questionnaire (DAQ: Clark et al., 1996). DESIGN, SETTING, AND PARTICIPANTS: Both questionnaires were administered, in a counterbalanced order, to a total of 380 recreational drinkers. In a further study, a shortened version of the DAQ was administered to a sample of 131 drinkers attending AA or a treatment centre. Exploratory factor analyses were carried out on the data and relationships between questionnaire score and other variables were assessed. FINDINGS: In recreational drinkers both instruments appeared to have a three-factor structure. The DAQ appeared superior to the ACQ in a number of respects: it produced more reliable factors; its structure accounted for a higher proportion of the variance; the factor inter correlations were somewhat lower; in a combined analysis of both instruments most of the factors retained came from the DAQ; and the DAQ discriminated better between bing and non-binge drinkers and excessive and moderate drinkers. A similar factor structure was found for the DAQ in the alcoholic subjects with addition of a factor of "controllability". CONCLUSIONS: The results support a multifactorial account of alcohol craving, and indicate that the DAQ has some advantages over the ACQ as a research tool. PMID- 9744140 TI - Death by methanol poisoning in Brazil. PMID- 9744141 TI - Lipid-lowering therapy: guidelines, targets and the need for action. PMID- 9744142 TI - Once-daily budesonide: 400 micrograms once daily is as effective as 200 micrograms twice daily in controlling childhood asthma. PETITE Research Group. AB - One hundred and sixty seven children on 0-200 microgram/day of inhaled steroid with asthma symptoms and sub-optimal peak flow values (less than 90% of that predicted for their height) were randomly allocated either 400 microgram once daily (nocte with placebo o.m.) or 200 mircrogram twice daily of budesonide Turbohaler for 8 weeks. Bronchdilator usage and symptoms were reduced in both groups at 4 and 8 weeks compared with baseline. There was a significant increase within both groups in morning and evening PEF after 4 and 8 weeks. The increase in evening PEF after 8 weeks was greater in the once-daily group than in the twice-daily group but there were no other significant differences between the groups (morning: +24.6 l/min vs 15.2 l/min, p = 0.059; evening: + 19.7 l/min vs +8.31 l/min; p = 0.013). Budesonide Turbohaler 400 microgram once daily is therefore as effective as 200 microgram twice daily in achieving asthma control in children. PMID- 9744144 TI - Pulmonary tuberculosis and its therapy do not significantly affect thyroid function tests. AB - Basal thyrotropin, total thyroxine, total tri-iodothyronine and free tri iodothyronine were measured in 30 HIV-negative patients with focal pulmonary tuberculosis upon starting and after one and two weeks of isoniazid, rifampicin and pyrazinamide therapy. Rapid and statistically significant elevations in free tri-iodothyronine and total tri-iodothyronine were noted after the first and second week of treatment respectively. For most patients, however, hormone levels remained within normal limits throughout the study. Overall, the effects of pulmonary tuberculosis and of its therapy on the thyroid function parameters examined, in otherwise healthy individuals, appear to be minimal. PMID- 9744143 TI - A comparison of nisoldipine coat-core and felodipine in the treatment of mild-to moderate hypertension. AB - The efficacy and safety of nisoldipine CC and felodipine were compared in a multicentre, randomised, double-blind, trial in patients with mild-to-moderate hypertension (n = 229). Following a two-week placebo run-in period, patients were randomised to 16 weeks' active treatment with nisoldipine coat core (CC) 20-40 mg or felodipine 5-10 mg once daily. At week 16, a higher proportion of patients in the nisoldipine CC group were on low-dose therapy (51% vs 36%, p = 0.07). The proportion of treatment responders was 77.8% with nisoldipine CC and 66.5% with felodipine. The mean change from baseline in systolic/diastolic blood pressure was -18.8/-13.6 mmHg with nisoldipine CC and -17.4/-11.3 mmHg with felodipine. The most common adverse events included peripheral oedema and headache; neither treatment affected heart rate. Thus, nisoldipine CC and felodipine provide comparable antihypertensive efficacy. The adverse effects of both drugs are related to their vasodilator properties and are common to the class. PMID- 9744145 TI - Telephone follow-up: an extension of the surgical outpatient department. AB - A total of 112 surgical patients took part in telephone follow-up, conducted by non-medical staff, after discharge home. Successful telephone follow-up was achieved in 93% of patients at 6 weeks, 91% at 3 months and 79% at 12 months. Telephone follow-up by non-medical staff is a practical method of assessing patient outcome after surgery. Wider use of this technique would free up time in the surgical clinics, without compromising audits of outcome and purchaser demands for evidence of efficacy. PMID- 9744146 TI - Incidence and prognostic implications of falls associated with acute medical illness: a medical inpatient study. AB - Falls are associated with a wide range of acute medical illness across all ages. They have prognostic implications, and screening for falls on admission can help identify those patients who are likely to have an adverse hospital outcome and may benefit from more intense levels of care. PMID- 9744147 TI - Growth and metabolic responses in preterm infants fed fortified human milk or a preterm formula. AB - Preterm infants fed human milk have been shown to grow poorly and develop mineral deficiencies that may lead to osteopenia. This study has investigated the efficacy of a human milk fortifier made up of glucose polymers, a mixture of bovine milk protein fractions and free amino acids, minerals and vitamins designed to improve these nutritional deficiencies. Growth and bone mineral deficiencies were compared in 38 preterm infants fed fortified mother's milk and 21 preterm infants fed a preterm formula until they reached 1800 g; all had a birthweight below 1600 g. Weight gain was similar in each group with a mean (SD) increase of 19.6 (3.5) g/kg/day in the fortified group and 19.9 (4.1) g/kg/day in the preterm formula group. There were also no significant differences in linear growth, head circumference, skin fold thickness or mid-arm circumference. Serum phosphate, alkaline phosphatase and plasma urea concentrations were similar and there was no clinical evidence of osteopenia. These results indicate that the growth and metabolic disadvantages associated with feeding human milk to preterm infants are ameliorated by the addition of a milk fortifier that increases the calorific, protein and mineral content of breast milk. PMID- 9744148 TI - The effect of dietary intervention on changes in total cholesterol, blood pressure and weight in a Cambridge study. AB - The study examined the impact of dietary advice on cholesterol reduction, to see whether diet was more effective when preceded by a cholesterol estimation. A total of 635 healthy individuals aged between 40 and 60 years attended a general practice cardiovascular risk screening in 1990. All subjects had their blood pressure, body mass index and smoking history recorded, and 50% were randomised to cholesterol measurement. Two years later, all individuals were invited to participate in a follow-up screening and 356 individuals took part (158 men, 198 women). All the above estimations were undertaken in 100% of attendees. The mean total cholesterol levels and weight declined in the subjects who were given dietary advice and had their cholesterol measured, while the mean cholesterol level increased slightly in subjects who did not receive dietary advice but had their cholesterol measured. The was a significant increase in diastolic blood pressure in all patients. The figures for reported smokers decreased from 22.4% to 19.4% in men and 19.4% to 8.4% in women. The results suggest that dietary advice can achieve a sustained reduction in cholesterol levels, but may require the stimulus of a prior cholesterol estimation. PMID- 9744149 TI - Management of vulval pain--whose responsibility? AB - Vulval pain can be acute or chronic. Usually patients present to their general practitioner when the pain is acute but, when specialist advice is required, vulval pain can present a real dilemma over to whom to refer patients. Genitourinary medicine, gynaecology, dermatology and sexual dysfunction clinics all play an important part in the holistic management of this symptom. Consultation in a sympathetic environment with appropriate investigations and collaboration between different disciplines may provide a diagnosis and management. PMID- 9744150 TI - Reversible left ventricular dysfunction: does it affect clinical practice and does it matter? AB - There is substantial evidence that many patients with impaired left ventricular function secondary to coronary artery disease may have hibernating or stunned myocardium. The identification of these patients is important, as revascularisation is associated with an improvement in function, and there is some evidence that revascularisation of these patients will actually improve prognosis. The most useful investigations for the identification of reversible left ventricular dysfunction are dobutamine echocardiography, thallium scanning and, although not available in many centres, PET scanning. PMID- 9744152 TI - The efficacy and safety of mibefradil in subgroups of patients with chronic stable angina pectoris. AB - The pool of controlled clinical studies on mibefradil, a new selective T-type calcium channel blocker, for the treatment of chronic stable angina pectoris was analysed to determine the effects in subgroups of patients defined by age, gender, body weight and common co-existing conditions. Total exercise tolerance test duration increased similarity in all subgroups, both at 50 mg (range 8.9 17.3%; n = 383) and 100 mg mibefradil (range 23.6-30.8%; n = 235). The increases in time to onset of angina and 1 mm ST-segment depression were similarly comparable. Safety and tolerability was similar to placebo for subgroups at 50 mg, but the incidence of adverse events was slightly higher in females, older, and lower weight patients at the 100 mg dose, particularly in reported leg oedema (7.9% in females vs 1.0% in males and 5.1% in older vs 0% in younger patients). Mibefradil proved a safe, well tolerated, and effective antianginal agent that can be used regardless of demographic factors or of frequently coexisting clinical conditions. PMID- 9744153 TI - Hypomagnesaemia in postoperative patients: an important contributing factor in postoperative mortality. AB - Magnesium is the second most abundant intracellular cation and the fourth most abundant cation in the body. Clinical manifestations of hypomagnesaemia include neuromusclar, neurological, psychiatric and cardiac arrhythmias including torsade de pointes resulting in sudden death. Incidence of hypomagnesaemia in hospitalised patients in common and there is a lack of clinical awareness. Clinicians should become familiar with the common conditions and therapeutics that are risk factors for underlying hypomagnesaemia and become familiar with magnesium replacement regimens. Two patients who suffered fatal complications in whom hypomagnesaemia was an important contributing factor are presented. Hypokalaemia and hypocalcaemia are common in severe magnesium deficiency and require concurrent monitoring and correcting. PMID- 9744151 TI - Biochemical markers of bone turnover. AB - Since bone markers may reflect different aspects of bone disorders and cell function, and osteolytic and osteoblastic activities may be individually or concomitantly altered, determination of more than one marker type is generally appropriate. Also, the individual markers of a particular type do not necessarily show parallelism. For example, in osteomalacia from vitamin D deficiency, bone specific alkaline phosphatase may be grossly elevated because of enhanced osteoblastic activity, whereas the vitamin D dependent osteocalcin may be decreased. With the exception of measurement of the bone enzymes, bone-specific alkaline phosphatase and tartrate-resistant acid phosphatase, bone marker measurements require complex and expensive immunoassays. As a general rule, the simple enzyme measurements can precede other investigation in most bone disorder> Bone-specific alkaline phosphatase measurement alone is generally adequate for the investigation of osteomalacia, Paget's disease and hyperparathyroidism but should be combined with measurement of tartrate-resistant acid phosphatase in suspected metastatic disease, and in multiple myeloma. Determination of both enzymes together may also be of value in the investigation of osteoporosis but in this disorder added benefit may be obtained by the addition of other bone markers, particularly urine deoxypyridinoline and possibly serum collagen telopeptide. PMID- 9744154 TI - Congenital double uterus and conglutinatio cervix in labour. AB - An interesting case of congenital double uterus and the intrapartum events associated with it are described. In view of the rarity of the condition, its implication to the patient, her general practitioner and the obstetrician/gynaecologist is discussed, with special reference to contraception, cervical smear and operative surgical technique. PMID- 9744155 TI - Volvulus of the right colon in pregnancy. AB - Intestinal obstruction in pregnancy is rare, volvulus being the commonest precipitating factor. It is a difficult diagnosis to make, because its symptoms mimic various pregnancy symptoms, but it commonly occurs in the third trimester or puerperium. A high index of suspicion is needed, and investigations for intestinal obstruction should be performed, especially when abdominal pain and vomiting persist. Careful evaluation of the abdominal cavity is also essential where a caesarean section is performed, particularly if there are unusual findings. PMID- 9744156 TI - Chicken fancier's spleen. AB - Splenic epidermoid cysts are rare congenital lesions which usually present insidiously with non-specific symptoms such as dull left upper quadrant pain, or as incidental findings on clinical examination. We present a chicken breeder who presented as an emergency with a tender left upper quadrant mass and septicaemia secondary to zoonotic infection of a primary splenic cyst with Salmonella enteritidis. The cystic nature of the swelling was confirmed by ultrasound and the anatomy assessed with computed tomography. She was aggressively resuscitated and underwent laparotomy and splenectomy, after which she made an uncomplicated recovery. The importance of early diagnosis and surgical treatment is discussed, together with the measures required as prophylaxis against overwhelming post splenectomy sepsis. PMID- 9744157 TI - Acute on-chronic middle ear disease. AB - The discharging ear is a common presenting symptom to GPs and otologists. The commonest causes are otitis externa and otitis media. Less common is cholesteatoma, which presents with a chronic offensive mucopurulent discharge and deafness. Acute mastoiditis as a presenting symptom of cholesteatoma is rare and, with the introduction of antibiotics, the incidence has declined even further. We present three cases of cholesteatoma presenting as mastoiditis. PMID- 9744158 TI - Cerebellar encephalitis following Epstein-Barr virus infection. AB - A case is described of acute cerebellar symptoms as the presenting features of Epstein-Barr virus (EBV) infection. Cerebellar encephalitis is discussed with particular reference to EBV infection. PMID- 9744159 TI - Profound hyponatraemia following an idiosyncratic reaction to diuretics. AB - A case report of a79-year-old woman with a breast adenocarcinoma who presented with clouding of consciousness and anorexia with a hyponatraemia of 97 mmol/l is described. An initial diagnosis of secretion of inappropriate antidiuretic hormone (SIADH) was made on the clinical history and a urine: plasma osmolality ratio of 2.25. Further investigation revealed the correct diagnosis of a sodium losing state with dehydration, and that her ADH level was normal. The actual cause of her hyponatraemia was the recent prescription of a thiazide diuretic. This case illustrates the unreliability of urine: plasma osmolality ratios for the diagnosis of SIADH. PMID- 9744160 TI - A survey of research in telemedicine. 2: Telemedicine services. AB - This article deals with research which attempts to demonstrate the value of health care delivered via telemedicine. A great variety of telemedicine services have been proposed. Much of the published work concentrates on the power of the technology and the content and quality of interaction it supports. There are other parameters to be considered in assessing telemedicine as a component of a health-care system, parameters which in practice determine the nature of the service being provided. Despite the fact that much work in telemedicine is published as short accounts of pilot projects, there is little research that explicitly addresses the difficulties involved in setting up new telemedicine services. Some studies have been performed which attempt to evaluate telemedicine services, but relatively few studies have been able to collect the data required to provide an unambiguous demonstration of their value. PMID- 9744161 TI - The legal and ethical aspects of telemedicine. 3: Telemedicine and malpractice. AB - This paper reviews the difficulties raised by the need to obtain valid consent from telemedicine patients to the treatment that they receive and evaluates the legal principles that the courts will apply to an allegation of negligence brought against a teleconsultant or telemedicine service by a patient. While many of the processes that take place during a telemedical consultation will be unique, it is suggested that the legal principles that apply to the conventional, face-to-face, doctor-patient relationship are equally as valid in the context of the practice of medicine at a distance. PMID- 9744162 TI - Telecare at home: factors influencing technology choices and user acceptance. AB - Acceptance of telecare technologies in the home is strongly influenced by factors that relate to dwelling design and the ways in which independent living is promoted. The experience to date of personal response systems (and services) provides some indication of the reasons why users may either resist their introduction or not use such technologies after they have been introduced. Telecare systems and services are poised to incorporate medical and other sensors, so that the manner of their provision, installation and control becomes an important consideration for service providers. A clear ethical framework is called for. PMID- 9744163 TI - A cost measurement study for a tele-oncology practice. AB - The costs of providing oncology services in three different ways were measured. Services were provided to a peripheral hospital by: conventional clinics, in which the oncologist worked at the hospital concerned; outreach clinics, in which an oncologist was flown in periodically from a central hospital; telemedicine clinics, in which the oncologist at the central hospital practised via a video link. During a one-year study period, 2400 patients were seen in conventional clinics, 81 in outreach clinics and 103 in telemedicine clinics. At these workloads the average costs per patient were $149, $897 and $812, respectively. However, the average costs cannot be compared directly without further information about the shape of the unit cost curves. PMID- 9744164 TI - A remote collaboration system for telemedicine using the Internet. AB - We have developed a still-image telemedicine system for the Internet. It was implemented with the Java programming language and enables remote collaboration between two or more client computers located anywhere on the Internet. Each client requires only a PC or workstation and a popular Internet browser--no special hardware or software is required. We carried out both local-area and wide area tests of the system. On a local network, JPEG images at a resolution of 640 x 480 pixels took 2-5 s to display on four different clients (PCs or workstations); at 1000 x 1000 pixels, the images took 10-65 s. We also tested the system between two universities in Japan, one in Italy and one in the USA, using the Internet for communications. Images required 4-20 s for display. The exchange of remote collaboration commands between the four locations was good and the maximum lag in mouse pointer movement was less than 1 s. The system has the potential to solve three problems of conventional telemedicine systems: their cost, the need for high-bandwidth telecommunication and the low compatibility between them. PMID- 9744165 TI - Diagnostic accuracy and clinical management by realtime teledermatology. Results from the Northern Ireland arms of the UK Multicentre Teledermatology Trial. AB - Diagnostic accuracy and management recommendations of realtime teledermatology consultations using low-cost telemedicine equipment were evaluated. Patients were seen by a dermatologist over a video-link and a diagnosis and treatment plan were recorded. This was followed by a face-to-face consultation on the same day to confirm the earlier diagnosis and management plan. A total of 351 patients with 427 diagnoses participated. Sixty-seven per cent of the diagnoses made over the video-link agreed with the face-to-face diagnosis. Clinical management plans were recorded for 214 patients with 252 diagnoses. For this cohort, 44% of the patients were seen by the same dermatologist at both consultations, while 56% were seen by a different dermatologist. In 64% of cases the same management plan was recommended at both consultations; a sub-optimum treatment plan was recommended in 8% of cases; and in 9% of cases the video-link management plans were judged to be inappropriate. In 20% of cases the dermatologist was unable to recommend a suitable management plan by video-link. There were significant differences in the ability to recommend an optimum management plan by video-link when a different dermatologist made the reference management plan. The results indicate that a high proportion of dermatological conditions can be successfully managed by realtime teledermatology. PMID- 9744166 TI - Staff perceptions of emergency and home-care telemedicine. AB - The utilization of a low-bandwidth telemedicine system for emergency and for home care was studied in a pilot trial. The emergency setting was the emergency department of a small urban hospital and its emergency medical service (EMS); the home-care setting was the home-health agency affiliated to the hospital. Utilization data were obtained through baseline and follow-up interviews with EMS technicians, emergency department and home-health nurses, and the project coordinator. The results indicated that initial enthusiasm for the use of the telemedicine system was not followed by a commitment to the system's utilization during the trial by the relevant administrations. Barriers to optimum utilization were identified, but the actual value of the system to patient care could not be determined. We conclude that the value of a telemedicine system to patient care cannot be realized unless there is an organizational commitment from the top to system utilization. PMID- 9744167 TI - A pilot trial of digital imaging in skin cancer. AB - We have used inexpensive off-the-shelf equipment for store-and-forward teledermatology and compared the precision and accuracy of digital image consultations with conventional, clinic-based consultations. Thirteen lesions were studied on 12 patients referred to a dermatology clinic for a suspected skin cancer. Patients were examined by two dermatologists. Subsequently, digital images were examined by two different dermatologists. There was almost complete agreement, both among and between the clinical and digital examiners, on different diagnosis and biopsy recommendations. Agreement on the single most likely diagnosis was also good. Digital imaging shows promise in teledermatology. PMID- 9744168 TI - A pilot study of tele-oncology in Scotland. AB - A pilot study of tele-oncology linking a cancer centre with a rural district general hospital was carried out; it involved patients, physicians, surgeons, radiologists and nursing staff. Its purpose was to complement the existing on site outpatient services, providing oncological advice on non-clinic days. During the six months of the trial, 18 videoconferences were conducted. Their median duration was 17 min (range 7-40). Eight videoconferences involved patients directly. Acceptability of videoconferencing to doctors, nurses and patients was assessed by a questionnaire. Patients and staff found the technique acceptable and were satisfied with the results. The addition of a teleradiology system to teleconsultations was found to be important when decisions on patient management were taken. Following the success of this pilot trial, large studies of tele oncology in the UK with measures of cost-effectiveness are needed. PMID- 9744170 TI - Remote psychotherapy. PMID- 9744169 TI - Telemedicine conference on a 13-year-old Chinese girl with an unusual skeletal condition. PMID- 9744171 TI - The state of public health research in New Zealand. PMID- 9744172 TI - The economics of public health. PMID- 9744173 TI - Enhancing NHMRC investment in public health research. PMID- 9744174 TI - The public health challenge of hepatitis C. PMID- 9744175 TI - Malaria and its implications for public health in Far North Queensland: a prospective study. AB - This prospective study's objectives were to describe the features of all episodes of malaria diagnosed in Far North Queensland (excluding the Torres Strait) and to assess how much of a threat they posed to the area's public health. Over a three year period, 216 episodes of malaria were diagnosed (158 Plasmodium vivax and 68 P. falciparum infections). Most (82%) of the infections were acquired in Papua New Guinea (PNG). Approximately 70% of the episodes occurred in Australian citizens, about half of whom were in malaria-endemic countries for work; the remainder travelled abroad for recreation. Three-quarters of the Australian citizens with malaria had taken either no or inadequate prophylaxis. Australian citizens who had taken adequate prophylaxis were much less likely to develop P. falciparum than other types of malaria compared to those who took either no or inadequate prophylaxis (p = 0.01). Gametocytes were present in 121 (56%) of the episodes of malaria. Mosquito surveillance was carried out in response to 38 (31%) of these gametocytaemic episodes. Significant numbers of Anopheles farauti sensu lato mosquitoes were found close to the residence of a patient in 4 (11%) of these episodes. Only two occasions when local transmission could have possibly occurred were recognised. We do not believe malaria poses an important threat to the health of the public in Far North Queensland. Nevertheless, it remains an important problem for those who travel abroad to malarious areas. PMID- 9744176 TI - Health promotion in different medical settings: women's health, community health and private practice. AB - This paper describes the health promotion role of doctors in two medical practice settings: women's and community health centres, and fee-for-service practice. It proposes the establishment of divisions of primary health care in Australia which would be multi-disciplinary and focus on community-wide health issues. The paper is based on data from an interview survey of medical practitioners who had worked in metropolitan Adelaide women's and community health centres and from a questionnaire survey of GPs in private practice. The types of health promotion activity by the doctors in the different settings are discussed. It is concluded that private practice GPs are involved primarily in providing health education advice to individual patients. Doctors within women's and community health centres are more likely to report involvement in group health promotion activity and broader community development initiatives. The study concludes that health promotion which focuses on the health of the local community is best conducted within multidisciplinary health centres. GPs in private practice are limited by the structure of their setting (particularly the fee-for-service basis and reliance on a single discipline) to health promotion which focuses on the needs of individual patients. PMID- 9744178 TI - Compliance with anti-tuberculosis preventive therapy among 6-year-old children. AB - There are no published data regarding compliance with anti-tuberculosis preventive therapy among children in Australia and limited published data worldwide. This study aimed to determine the compliance rate among 6-year-old children prescribed preventive therapy for tuberculosis infection. A prospective cohort study was conducted on 78, 6-year-old children prescribed antituberculosis preventive therapy. Compliance was measured by compliance with prescribed preventive therapy as reported by parents who were administered questionnaires on completion of the course. In a subsample of 44 children, the proportion of children who complied with scheduled visits to the hospital, and pharmacy records of isoniazid dispensed were used as measures of compliance. Questionnaire data indicated a reported compliance rate for completion of the 6-month course of preventive therapy of 70.5% (55 children). For those 55 who reported completing the full course, 91% reported missing less than 1 tablet per week. In the subsample of 44 children, only 59% attended all follow-up clinic visits, and 54% collected all 6 months of isoniazid prescribed. Compliance with antituberculosis preventive therapy is suboptimal. Improved methods to measure compliance and strategies to optimise compliance with preventive therapy is required. PMID- 9744177 TI - Effect of training on general practitioners' use of a brief intervention for excessive drinkers. AB - OBJECTIVE: To determine among general practitioners (GPs) the effect of three different types of training on utilisation of a brief, controlled drinking intervention. DESIGN: A non-randomised intervention study. Setting, participants: 96 GPs (64%) within the South Eastern Sydney Division of General Practice participated; 35 chose workshop training, 39 one-to-one training and 22 received a special kit by mail. MAIN OUTCOME MEASURES: Identification by GPs of excessive drinkers by practice audits; use of the program determined by the number of patients recruited in 3 months and by GPs' use of the intervention 6 months after training. RESULTS: 41 (43%) GPs conducted practice audits, identifying 15.1% of males and 6.6% of females as excessive drinkers (regular excessive weekly consumption and/or binge). 179 patients were recruited by 36 GPs over 3 months, and 32% of these patients reported a reduction of alcohol consumption. 63% who attended workshop training, 57% who received one-to-one training, and 36% who received the kit by mail reported they were current users of the program at 6 months. Significantly fewer GPs who received the kit by mail reported ever using the program (59%) compared to the other groups (p < 0.01). CONCLUSION: This naturalistic study found that workshops and one-to-one training sessions in doctors' surgeries achieved greater uptake of a brief intervention for problem drinkers than distribution of a special kit by mail. PMID- 9744179 TI - Admission rates as an indicator of the prevalence of severe asthma in the community. AB - BACKGROUND: A reliable indicator of the prevalence of severe asthma in the community is needed to monitor population-based asthma control strategies. We examined the potential use of asthma admissions to hospital as such an indicator. METHODS: We recruited subjects from the Emergency Department (ED) of a children's hospital. The attending doctor completed the 'physician questionnaire' which included questions on the patient's asthma severity and interval severity/chronicity of asthma. The parent/guardian completed the 'parent questionnaire'. It included questions on demography, asthma knowledge and attitudes, asthma history and social support. We performed univariate and multiple logistic regression to determine predictors for hospital admission. RESULTS: Interval severity of asthma, pre-treatment severity of wheeze and low post-treatment pulse oximetry best predicted whether children presenting with asthma were admitted. Demographic variables, factors associated with access to health services and factors related to the asthma history and management were not significant predictors of admission. DISCUSSION: At the population level, it may be possible to utilise routine hospital admission rates as an indicator of the prevalence of severe asthma in the community, especially within the context of monitoring trends in asthma prevalence. Our study was conducted in a metropolitan tertiary paediatric hospital. The reliability of hospital admission rates as indicators of the prevalence of severe asthma in other hospital settings, in different population groups and over time remains to be established. PMID- 9744180 TI - Use of focus groups to identify health promotion strategies for the early detection of diabetic retinopathy. AB - BACKGROUND: People with diabetes do not regularly utilise eye services for the early prevention of vision loss due to diabetic eye disease. A community-based screening program has been initiated in Victoria to address this issue. To encourage people to take preventive eye health care measures, the most effective health promotion strategies were identified. METHODS: Thirty-three health professionals were invited to attend focus groups. A sample of 35 people with diabetes was approached by their GPs or diabetes educators because of their motivation to participate in diabetes activities. Each group consisted of 10 members. Discussion points included the type of education messages available to people with diabetes; use of eye services among the participants with diabetes; and strategies required promoting the screening service. RESULTS: Five focus groups were conducted. The discussions highlighted that a great deal could be achieved by using local community networks to promote the benefits of early detection of diabetic retinopathy and local screening program. The group members recommended that particular attention be directed to general practitioners and their distribution of materials to patients. Key issues for planning and implementing the program were highlighted. The groups urged development of strategies to encourage people with diabetes in rural Victoria to participate in a program for the early detection of diabetic retinopathy. PMID- 9744181 TI - Differences in health estimates using telephone and door-to-door survey methods- a hypothetical exercise. AB - Telephone interviewing is increasingly being used to obtain data on health issues. Propertly applied telephone interviewing may have considerable cost benefits, but careful thought has to be put into the design of surveys, weighting and analysis of data to avoid major sources of bias. This study is a hypothetical exercise comparing health estimates from a systematic, self-weighting, multistage, clustered, area sample of households using a face-to-face interview method, with hypothetical samples of people obtained from Random Digit Dialling and Electronic White Pages. In a comparison of the population health estimates obtained for a number of health problems in a hypothetical analysis of these samples, the confidence intervals for the estimates overlapped. Since the estimates are not statistically significantly different, it appears that well planned, appropriately weighted and analysed telephone surveys can be a less expensive way of obtaining health information, however, some caution is expressed in using this method. PMID- 9744182 TI - Mental health services in Perth nursing homes. AB - Directors of nursing (DONs) in 42 nursing homes in Perth were asked for information concerning mental health services provided for their residents. A questionnaire similar to those used by other researchers in Sydney and in Ontario was utilised to enable meaningful comparisons among the three studies. According to the DONs' perceptions, the mean proportion of residents in Perth nursing homes with psychiatric and/or behavioural problems was between 50% and 75%. The mental health services provided to nursing homes were significantly less than that desired by the DONs. Other variables assessed in this study included the number of transfers per year because of psychiatric problems; the number of staff with psychiatric qualifications; the provision of psychiatric training for staff; the most frequently occurring psychiatric problems in residents and the most valued mental health services provided to nursing homes. Results were comparable to those of the Sydney and Ontario studies. This study suggests that there is a significant psychiatric population in Perth nursing homes that, like those in Sydney and Ontario are seriously neglected regarding appropriate professional psychiatric care. Existing psychogeriatric community assessment teams could provide more of these services if adequately staffed and resourced. PMID- 9744183 TI - Does breastfeeding at six months predict cognitive development? AB - There is controversy over whether the method of feeding in infancy affects intellectual development. We investigated the relationship between breastfeeding status at 6 months of age and long-term cognitive development in a cohort of 375 children born in Port Pirie, South Australia, between 1979 and 1982. Cognitive assessments were conducted at ages 2, 4, 7 and 11 to 13 years. After adjustment for sociodemographic, environmental and biomedical factors, a small, statistically non-significant, beneficial effect of breastfeeding on cognitive functioning was observed. Compared with the bottle-fed children, the breast-fed children had a 3.4 (95% CI -0.1 to 6.9), 1.3 (-2.3 to 4.9), 1.2 (-2.0 to 4.4) and 0.8 (-1.9 to 3.5) point advantage on the Bayley Mental Developmental Index at age 2 years, the McCarthy General Cognitive Index at age 4 years and the Wechsler Full-Scale IQ at ages 7 and 11 to 13 years, respectively. Our data suggest that any beneficial effect of breastfeeding on cognitive development is quite small in magnitude. PMID- 9744184 TI - Consumption of different alcoholic beverages as predictors of local rates of night-time assault and acute alcohol-related morbidity. AB - OBJECTIVE: To determine whether population levels of consumption of some alcoholic beverages are more closely associated with levels of harm than others, particularly if consumption of cask wine is more strongly related to rates of acute alcohol problems than consumption of bottled wine as a consequence of the extremely low rates of federal tax levied on the former. METHOD: A database of alcohol consumption and related problems was established for 130 areas of Western Australia. Demographic and economic data for these areas were included from the 1991 census. Empirically derived assumptions regarding the mean wholesale price of cask and bottled wine were utilised. Regression analyses examined the extent to which the consumption of different alcoholic beverages predicted levels of major varieties of harm. RESULTS: Only cask wine and high-strength beer consumption were significantly associated with rates of night-time assault; consumption of all beverage varieties except bottled wine was significantly associated with rates of acute alcohol-related morbidity. Further analyses, which included controls for an effect of total alcohol consumption, confirmed the pronounced contributions of cask wine and high-strength beer to rates of night assaults and acute alcohol-related morbidity. The proportion of all alcohol consumed as low-alcohol beer was significantly negatively associated with these harms. CONCLUSIONS: The beverages most associated with rates of night-time assaults and acute alcohol-related morbidity are those with the lowest federal taxation per standard drink, i.e. cask not bottled wine and regular-strength not low-alcohol beer. PMID- 9744185 TI - A salmonellosis outbreak linked to internally contaminated pork meat. AB - In August 1995, we investigated an outbreak of salmonellosis among patrons who attended a church camp in southern Sydney. Of the 73 attendees interviewed, 22 reported a gastroenteritis illness within two days of the conclusion of the camp, with one attendee hospitalised. Two stool specimens, one from each of two attendees, were both positive for Salmonella typhimurium phage type 9. A cohort study of 68 attendees established a statistically significant association between illness and the consumption of de-boned roast pork (estimated relative risk infinite, p = 0.03) and between illness and the degree of cooking of the pork meat (chi 2 for trend 5.8, p < 0.02). The outbreak was most likely caused by consumption of roast pork that had been internally contaminated during the de boning process. Meat and meat products that may be internally contaminated, such as de-boned meats, should be thoroughly cooked. Guidelines about minimum cooking temperatures of meats liable to internal contamination should be developed for commercial food handlers in Australia. PMID- 9744186 TI - Economic evaluation of health promotion: friend or foe? AB - It is commonly believed that economic evaluation is hostile to health promotion and that the requirement for health programs to be cost effective will result in a biased allocation of funds in favour of programs that can demonstrate short-run benefits as defined by inadequate outcome measures. The paper is concerned with the validity of this perception. It is argued that economic evaluation has the potential for treating health promotion activities on an equal basis with other health interventions. The major obstacle to this does not arise from the theory of economic evaluation, which is discussed, but from a lack of information about outcomes. Without this information any evaluation--economic or otherwise--is flawed. Three problems relating to the economic evaluation of health promotion activities are considered. These are: the discounting of future health benefits; the potential for economic evaluation to be counter-productive if applied to 'immature' projects; and the practical problems encountered in the measurement of the outcomes of health promotion programs. A four-fold classification which is based upon a distinction between disease cure, individual health promotion, community welfare and systemic change designed to promote either individual health or social well-being. The capacity of economics to incorporate these objectives is discussed. PMID- 9744187 TI - Postcard reminders from GPs for influenza vaccine: are they more effective than an ad hoc approach? AB - All persons 65 years and older are recommended to be immunised against influenza each autumn. As immunisation rates remain low, we conducted a randomised control trial in a three-partner urban general practice to evaluate the differential effectiveness of a single postcard reminder in a general practice setting compared to usual care. All non-residential patients aged 65 years and over were identified from the age/sex/disease register. After exclusions, 325 patients were stratified by sex (125 men and 200 women) and randomised to receive either a postcard reminder in large print mailed in April or usual care. General practitioners (GPs) were blind to the randomisation. A blinded record audit performed in July demonstrated that the postcard was effective in increasing immunisation for men (chi(2)1df = 3.85; p = 0.05) but not for women (chi(2)1df = 0.45; p = 0.50). After adjusting for 1995 immunisation status, the effect of the postcard on immunisation rates was even stronger in men (Wald chi(2)1df = 6.20; p = 0.01) but remained non-significant in women (Wald chi(2)1df = 1.38; p = 0.24). With this adjustment, the odds of having the 1996 flu vaccine for men sent the postcard reminder were three times that of men in the control group (OR = 3.0; 95% CI 1.3-6.9). In a general practice setting, a single postcard reminder appears to be a promising way to boost influenza immunisation rates among ageing men. Replication of the study is recommended. PMID- 9744188 TI - The burden of infectious disease in New Zealand. AB - New Zealand mortality records for the years 1980 to 1993 were analysed to estimate the aggregate burden of infectious disease using a recoding of ICD-9 codes to identify deaths with infectious aetiology. The recoding scheme was modified from one developed by US CDC, which used expert panels to assign ICD codes to categories dependent on the proportion of the code attributable to infection. ICD-9 Chapter One ('Infectious and parasitic diseases') accounted for only 0.7% of total deaths. Following recoding, this proportion increased tenfold, with 6.9% of deaths attributable to infectious disease. This proportion was stable or declined only slowly between 1980 and 1993. While rates varied by age, gender and ethnicity, the results indicate that infectious disease still accounts for a substantial proportion of the burden of disease in New Zealand. PMID- 9744189 TI - Infant hearing screening: a comparison of two techniques. AB - Hearing screening programs for Australian children are known to have poor coverage in many areas. In addition, only a minority of children are screened for hearing loss before 2 years of age. However, early detection of hearing loss and early treatment are generally considered very important to successful rehabilitation outcomes. Traditional methods of screening infants have limitations with their accuracy in detecting children with hearing loss. This study compared the results obtained with a traditional questionnaire approach to screening and a newer objective technique involving otoacoustic emission measures. Poor correlation was found between pass rates for the two techniques, suggesting that the questionnaire approach is not an accurate screening method for detecting infant hearing loss. With further development, otoacoustic emission testing holds promise as an objective alternative hearing screening procedure. PMID- 9744190 TI - The dietary intake of chemical residues in Brisbane adults. AB - The purpose of this work was to integrate existing chemical residue and food consumption data for individuals to improve estimates of the dietary intake of chemical residues in the population of Brisbane. Previous estimates of intakes from the Australian Market Basket Survey (AMBS) have been based on energy adjusted 'hypothetical national diets' and so allow no assessment of variation in intakes between individuals or groups. Data on concentration of fenitrothion, chlorpyrifos-methyl, pirimiphos-methyl, heptachlor and dieldrin in selected foods were taken from reports of the AMBS. Food consumption data were based upon the National Dietary Survey of Adults (NDSA) 1983; the same data from which the hypothetical diets are derived. The distribution of estimated 24-hour intakes was adjusted to represent usual intakes. Mean intakes of all residues were about one third those reported previously. None of the observed diets contained levels of residues that were greater than the Acceptable Daily Intakes. These findings support reassurances to the public that residues of agricultural chemicals monitored in the AMBS do not pose a health risk. PMID- 9744191 TI - Voluntary euthanasia: responses of medical practitioners in NSW and the ACT to six open-ended questions. AB - Six open-ended questions from a survey of 1271 randomly selected medical practitioners on active voluntary euthanasia (AVE) and physician-assisted suicide (PAS) was examined. In spite of some extreme written views for and against the procedures, the majority of practitioners were considered, concerned, sympathetic and troubled about AVE and PAS. PMID- 9744192 TI - Health funding: the nature of distortions and implications for the health service mix. AB - With restricted public sector budgets, there is increasing pressure to obtain value for money in the allocation of health sector resources. Distortions and inefficiencies created by features of Australia's health funding arrangements prevent health resources moving from lower valued to higher valued activities. These distortions tend to restrict resources being allocated to disease prevention and health promotional approaches, favouring medical treatment. These propositions are illustrated, largely with reference to diabetes. PMID- 9744193 TI - Vaccine storage in pharmacies on the Central Coast of New South Wales. AB - Pharmacies on the Central Coast of NSW were surveyed to assess the ability of refrigerators used for vaccine storage to maintain the recommended temperature range (2 to 8 degrees C). Refrigerators used for vaccine storage were monitored over a 3-day period using a temperature data logger. Fifty-nine (59) retail pharmacies were identified. The response rate was 90% (53/59) and 52 refrigerators were monitored successfully. Only 10 (19%) of the refrigerators maintained temperatures wholly within the recommended range for the 3-day monitoring period. The remaining refrigerators were considered in three groups- refrigerators keeping temperatures in the range 0.1 to 11.9 degrees C, below 0 degree C and above 8 degrees C for most of the time. There were 15 (29%), 12 (23%) and 15 (29%) refrigerators in these groups respectively keeping temperatures in the recommended range for 91%, 19% and 30% of the time. Of the refrigerators achieving temperatures below 0 degree C, none went below -5 degrees C and on average they kept temperatures less than 0 degree C for 49% of the time. This survey highlights the need for vigilance in vaccine storage for immunisation programs to be successful. Of the vaccines affected by freezing, hepatitis B vaccine was identified as being most at risk. PMID- 9744194 TI - Nurses' attitudes to active voluntary euthanasia: a survey in the ACT. AB - National public opinion polls show a large majority of Australians are in favour of active voluntary euthanasia (AVE). However, most members of the public have had only limited direct experience with dying people. For this reason, surveys of the opinions of medical practitioners and nurses on this issue are of great interest. The present study involved a postal survey in late 1996 of 2,000 randomly selected registered nurses from the Australian Capital Territory (ACT). The ACT has had extensive public debate about this issue. The questionnaire included some questions asked in earlier Australian surveys of the general public and health practitioners. Responses were received from 1218 nurses (61%). A majority of nurses who responded supported AVE as 'sometimes right', be it homicide by request (72%) or physician-assisted suicide (71%). A slightly smaller majority believed the law should be changed to allow homicide by request (69%) and physician-assisted suicide (67%). If AVE were legal, 66% of the nurses indicated they were willing to be involved in the procedure. Only 30% were willing to assist patients to give themselves the lethal dose, while 14% were willing to administer the lethal dose to the patient. Comparing these results with previous surveys, it appears that nurses are less in favour of AVE than the public, but more in favour than medical practitioners. PMID- 9744195 TI - Are cuts to health expenditure in Victoria compromising quality of care? AB - Separate state-wide surveys of women who had recently given birth in Victoria were conducted in 1989 and 1993. The first survey was conducted in conjunction with the Victorian Ministerial Review of Birthing Services. The second survey occurred three years after the release of the Review's final report and three months after the introduction of casemix funding. It coincided with a period of substantial cutbacks to expenditure on Victorian public hospitals. In both studies, surveys were mailed to women 6 to 9 months after the birth. Response rates were 71.4% (n = 790) in 1989 and 62.5% (n = 1336) in 1993. Between the two surveys, the proportion of women giving critical feedback about caregivers, more than doubled. The survey findings suggest that standards of care are being compromised in the current economic and policy environment. PMID- 9744196 TI - Panis populi--bread and public health in Australia. AB - The 'standard loaf, 680 gm, white, supermarket-purchased' as expressed in the Consumer Price Index, is but the basic form of bread sold to the Australian public. In the public health context, three themes have been intimately associated with bread--quality control, price control and bread used as a vehicle for supplementary nutritive agents important in preventive medicine. Price control, through assizing, has been a feature of bread marketing in western communities for seven centuries; and bread remains the last item on which price control (although seldom enforced) exists in Australia. Quality control, for public health, is determined both by regulation and by the force of increasingly literate consumers, of whom women occupy the most important determinant. From the preventive medicine point of view, important themes in bread quality, such as its use to reduce laxative sales on the one hand and to reduce the demographic incidence of colonic cancer on the other, remains outside formal regulation. Australia is a relatively conservative nation in the context of nutritional additives. It was not until 1953 that the National Health and Medical Research Council approved the addition of extra B vitamins to bread. Currently, folic acid is added as a discretion to selected high-premium breakfast cereals in Australia in one attempt to reduce the incidence to neural tube defects. The addition of such ingredients to bread remains an unrealised, but potentially important aspect of preventive medicine in Australia. PMID- 9744197 TI - Moving child health outcomes forward. PMID- 9744198 TI - Need to review guidelines for prevention of choking on food in children. PMID- 9744199 TI - Hazards of decontextualised accounts of public perceptions of radio frequency radiation (RFR) risk. PMID- 9744201 TI - Lyme disease in Australia. PMID- 9744200 TI - Smoking and pregnancy: time to implement evidence-based solutions. PMID- 9744202 TI - Needs assessment of rural and remote women travelling to the city for breast cancer treatment. AB - The purpose of this study was to assess the needs of rural women travelling to the city for breast cancer treatment. Participants included 80 women aged between 34 and 80 years living in rural NSW and South Australia who travelled for breast cancer treatment. After completing treatment, participants completed a brief telephone survey on the needs of rural women travelling for treatment. Findings revealed that more than 90% of women travelled for treatment due to the lack of available treatment centres closer to home and on average they spent 6.79 weeks (SD = 4.73) away from their home and family. Findings also showed that 89% identified specific problems for rural women, with social and practical support being primary concerns. Although the majority of women were provided with some type of social support, only 39% of women received financial assistance and 19% of these women had trouble claiming money for which they were eligible. Recommendations of appropriate interventions to ensure equity in the availability and access to breast cancer treatment for all women are discussed. PMID- 9744203 TI - Focus group interviews examining attitudes to randomised trials among breast cancer patients and the general community. AB - OBJECTIVE: To explore the knowledge of, and attitudes towards, randomised clinical trials among women in the community and breast cancer patients. DESIGN: Focus group interviews were conducted with women in the community and women previously treated for localised breast cancer. PARTICIPANTS: Twenty one mothers or grandmothers of children attending a local primary school and 20 breast cancer patients identified from the records of the Medical Oncology Department, Royal Prince Alfred Hospital, participated in one of eight focus group discussions examining knowledge of and attitudes towards randomised clinical trials. RESULTS: Most women did not have a good understanding of the need for clinical trials and the manner and safeguards with which they are conducted. They did not understand the need for randomisation and were often confused about the use of placebos. Many women were wary about medical research and saw it as a gamble, only to be considered if all else failed. Clinical trials were felt to be of benefit to future generations and perhaps family members if they should fall ill. However, they were not thought to be of benefit to the individual patient. CONCLUSIONS: These results suggest that greater community awareness of clinical trials is needed to improve participation in clinical trials. These focus group findings require validation in a larger sample. PMID- 9744204 TI - HIV positive tests at Coronial Services in Victoria 1989-1996: lessons for HIV surveillance. AB - Results of routine testing at other sites can supplement surveillance of the HIV epidemic in Australia which is largely based upon voluntary testing. Since 1989, systematic onsite HIV testing has been undertaken on all bodies taken to the Victorian Institute of Forensic Medicine (VIFM). Information was collected on all cases of HIV infection detected at VIFM between 1989 and 1996, and matched to surveillance databases. In 8 years, 75 people were diagnosed with HIV; all except one were male. The age range was 14-70 years, mean 37.4 years. The major causes of death were suicide 35%, AIDS 21%, drug toxicity 16%, natural causes 12% and injury 7%. The major exposure categories were male homosexual 51%, male bisexual 11%, homosexual/bisexual IDU 16%, IDU other 8%, and haemophiliac 7%. For only two was exposure information unavailable. Seropositivity for anti-HCV and HBsAg was 37% and 11% respectively. The deceased was recorded as having HIV/AIDS on the police report in 73% of cases, and at least 90% of subjects had been diagnosed with HIV prior to their death. The study suggests there is relatively little undiagnosed HIV infection in Victoria, that HIV infection has not moved outside traditional risk groups, and that many tests for HIV are undertaken using false namecodes. Many patients could not be matched on the HIV/AIDS databases, identifying a problem with HIV surveillance systems in Victoria, and the need to capture all information on HIV positives detected at VIFM. PMID- 9744205 TI - A South Australian Salmonella Mbandaka outbreak investigation using a database to select controls. AB - Between April and June 1996, 15 persons with Salmonella enterica serovar Mbandaka infection were reported in South Australia (population 1.6 million) compared with 12 over the previous five years. To identify a possible source for the infections a case control study was conducted. METHODS: Trained interviewers asked 15 cases and 45 controls about their consumption of 105 foods. Controls were matched to case residential location and age. They were selected from a previously constructed database of 3,014 randomly selected South Australian households. RESULTS: Thirteen of the 15 cases ate 'generic' or 'retail store' brands of peanut butter produced by the same factory in another state, compared with five of the 45 controls (p < 0.01). Salmonella Mbandaka was isolated from three opened jars of peanut butter from case households, and from three unopened jars from retail outlets. Further investigation implicated roasted peanuts from a third Australian state as the source of the Salmonella contamination. DISCUSSION: This is the first recorded outbreak of salmonellosis resulting from the consumption of peanut butter. The SA outbreak investigation comprised a matched case control study to identify possible common food sources. Such investigations need be conducted rapidly to maximise public health benefits, and the utility of selecting controls from a population based database can improve timeliness. PMID- 9744206 TI - Quality of life and later adverse health outcomes in patients with suspected heart attack. AB - We tested the hypothesis that low quality of life (QOL) after discharge from hospital with ischaemic heart disease (IHD) is associated with higher rates of later adverse outcomes (death and subsequent hospital admission for acute myocardial infarction or congestive cardiac failure). Three hundred and seventy five patients previously enrolled in an intervention study which assessed QOL six months after hospitalisation were followed up for an additional 18 months. The rates of adverse outcomes increased as QOL decreased (high QOL 9%; moderate 18%; low 28%). After adjustment for known prognostic factors, the risk of an adverse outcome was still higher in 'low' and 'moderate' compared to 'high' QOL subjects (low QOL adjusted OR = 2.6, 95% CI = 1.2-5.8; moderate 1.9, 0.8-4.2). In conclusion, QOL after discharge from hospital appears to be an independent predictor of later morbidity and mortality. PMID- 9744207 TI - Palatability versus healthiness as determinants of food preferences in young adults: a comparison of nomothetic and idiographic analytic approaches. AB - Past research on adults has found that the sensory appeal or taste of foods is a primary determiner of food consumption and how people think about food. The nomothetic nature of this research may have underestimated the impact of health considerations on food choice. This study compared 'nomothetic' and 'idiographic' modes of analysis in 1) determining the relative influence of palatability and perceived healthiness of foods, on preference for the food, and 2) assessing the relationship between palatability and evaluations of healthiness. Additionally, gender differences were examined in relation to within-person correlations between the concepts of preference, palatability and healthiness. Subjects (n = 139) rated 81 foods on preference, palatability and healthiness. Findings from both the idiographic and nomothetic analyses indicated that palatability rather than health considerations determined preferences in young adults. The within person correlational analysis indicated a large number of persons, mostly female, who preferred unhealthy food. The sample was equally split in their evaluations of healthy food as palatable or not. PMID- 9744208 TI - A multi-state outbreak of Salmonella bredeney food poisoning: a case control study. AB - OBJECTIVE: To investigate a multi-state outbreak of Salmonella bredeney. DESIGN: Case interviews followed by an age and neighbourhood matched case control study. PARTICIPANTS: People with laboratory-confirmed S. bredeney and controls matched on age and geographical location in New South Wales, the Australian Capital Territory, Queensland and Victoria. RESULTS: We identified 157 persons with S. bredeney spread throughout the eastern states and the ACT. In the matched analysis, cold meat and chicken demonstrated a significant odds ratio of 4.4 (p = 0.017) and 4.2 (p = 0.02) respectively. Among primary cases, the odds ratio for chicken was 6.0 (p = 0.01) and for ground pepper was 3.75 (p = 0.04). CONCLUSIONS: The most likely source of this outbreak was a product contaminated at the point of manufacture and distributed widely within NSW and the ACT and, to a lesser extent, Queensland (Brisbane) and Melbourne. The most probable food is a meat or chicken product, followed by substantial cross contamination of other meat products at retail outlets, which served to amplify the outbreak. PMID- 9744209 TI - Air pollution in the Latrobe Valley and its impact upon respiratory morbidity. AB - OBJECTIVE: To assess the relationship between air pollution and respiratory morbidity. DESIGN: An ecological study of the daily hospital admissions abstracted for the 1988 calendar year. Air quality data, including nitrogen dioxide (NO2), sulphur dioxide (SO2), ozone (O3) and particulates, were obtained from the relevant authorities. SETTING: Latrobe Valley, Victoria. SUBJECTS: Hospital admissions for asthma and Chronic Obstructive Airways Disease. (COAD). RESULTS: There were significant associations (r = 0.11 to 0.17) between airborne particles, nitrogen dioxide and respiratory morbidity. There was no significant relationship between any of the pollutants and asthma admissions. However, multi variate analysis confirmed that NO2 and particulates were associated with admissions for COAD. CONCLUSION: Respiratory morbidity appears to be affected even by the low air pollution levels in the Latrobe Valley. PMID- 9744210 TI - Are young adults checking their skin for melanoma? AB - This study examined the prevalence and predictors of self screening for melanoma in a large sample of young New Zealanders. A self-report questionnaire was administered to a sample of 909, 21-year-olds to investigate if young adults check their skin for changes in lesions which could be melanoma, and to identify the factors which influence this behaviour and any subsequent help seeking. Fifty three per cent reported checking their skin in the past year, with 20% noticing a change in a mole or freckle. Forty-five per cent of those who noticed a change sought medical advice. The most common reason for not seeking advice was cost. Women were more likely that men to have checked their skin, to have noticed a change and to have sought medical advice. In addition to gender, tendency to self check was also associated with knowledge of melanoma and perceived risk of melanoma. These results are discussed in light of the current debate regarding skin cancer screening. This study fills a gap in the literature regarding self screening for melanoma in young adults and identifies ways in which future prevention campaigns might be modified to address the concerns of this age group. PMID- 9744211 TI - A methodology for sampling and accessing homeless individuals in Melbourne, 1995 96. AB - A methodology for sampling homeless populations in inner Melbourne was developed to study their health status and prevalence of tuberculosis. This paper describes the design, development and implementation of the project. The results of health status and tuberculosis analysis are published elsewhere. Involvement and interaction with local service providers and agencies to homeless people was central to the project throughout. A definitional construct of homelessness was developed, drawn from local and overseas literature and contemporary local experience. The study's aim was to obtain a representative sample of homeless individuals in various levels of accommodation and a convenience sample of those who were unaccommodated (streets and parks). A comprehensive sampling frame of accommodation options was constructed from available databases, and systematic sampling applied to produce a sample of 396 beds, from which 284 participants were enrolled. Convenience sampling of unaccommodated homeless individuals produced 100 participants. All agreed to undergo a comprehensive questionnaire, blood and Mantoux testing, the latter being completed successfully in 94%. Commonsense, cultural sensitivity and a non-threatening approach were critical to the success of the project and the security of the field workers. The methods described attempt to address recognised difficulties of sampling from homeless populations and should be reproducible both in the future and elsewhere. Potential for selection bias remains the main threat to validity, which the described methodology combined with adequate resources should help to address. PMID- 9744212 TI - Serostatus for vaccine-preventable diseases in residents at Melbourne Juvenile Justice Centre. AB - There are concerns in Australia about inadequate rates of childhood immunisation, an important preventive measure to reduce infectious diseases. The population passing through the Melbourne Juvenile Justice Centre (MJJC) comes from a background at high risk for inadequate immunisation and outbreaks can occur in residential institutions. MJJC residents were invited to participate in a study by completing a medical officer-administered risk behaviour questionnaire and/or giving a blood sample. Ninety residents completed the questionnaire; 83 gave blood samples. Sera were tested for measles, mumps, rubella and hepatitis A, B and C markers using standard commercial assays. Diphtheria and tetanus were tested with an ELISA in a public health laboratory familiar in the technique. Sixty four per cent (53/83) of participants were non-immune to at least one component of MMR and 44.6% (37/83) non-immune to either tetanus or diphtheria. Despite 61.1% of participants reporting injecting drug use, only 28.2% had protective levels of anti-HBs, 6.1% were positive for anti-HBc (2.4% equivocal), 22.9% were anti-HCV positive and 9.6% had markers of exposure to hepatitis A virus. These results show suboptimal levels of immunity in this institutional setting with the potential for disease outbreaks. Many residents miss adolescent school-based programs for immunisation because of truancy and early school leaving. Despite considerable risk of blood-borne viruses, many MJJC residents are inadequately vaccinated against hepatitis B. PMID- 9744213 TI - A prospective cohort study of blood donors: methodological issues in the investigation of injuries and chronic diseases. AB - Blood donors have made important contributions to research, most notably in cross sectional seroprevalence studies. The proposed New Zealand Blood Donors Health Study is a prospective cohort study of 30,000 New Zealand donors designed to investigate the determinants of common injuries, cardiovascular disease and cancer. While robust from an analytic perspective, the execution of prospective cohort studies in many settings is impeded by methodological, economic and organisational barriers. We examined the operational considerations of implementing a large-scale cohort study at a transfusion centre and evaluated measures taken to optimise data collection procedures. A pilot study of 1,000 participants revealed donor motivation to participate in this research was high (91% response rate). Comprehensive exposure data on lifestyle, behavioural and psychosocial factors were obtained from 95% of participants. Substantial heterogeneity in levels of potential risk factors was noted among respondents. Detailed dietary habit information and a study blood sample were obtained from 67% and 100% of participants, respectively. Study recruitment and baseline data collection was feasible during routine donor visits with minimal interruption to donor centre staff and procedures. We conclude the study design and characteristics of the regional donor program enhance the efficiency and significance of the proposed research. PMID- 9744214 TI - Pre-admission education/counselling for patients undergoing coronary angioplasty: impact on knowledge and risk factors. AB - This study aimed to evaluate whether a pre-procedural education/counselling program can improve knowledge and coronary risk factors in 130 patients (65 experimental; 65 comparison) approximately four months after having percutaneous transluminal coronary angioplasty (PTCA). Knowledge and physical activity levels (p = 0.00) improved for both groups from pre-PTCA to the follow-up. Further, the experimental group showed favourable change in total cholesterol level (p = 0.02) at follow-up. That participation in the intervention did not improve knowledge or risk factor prevalence may reflect the adequacy of standard ward care, the influence of factors not under the control of the study or the overall experience of PTCA. Limitations of educational programs without follow-up sessions are discussed and alternate rehabilitation approaches suggested. PMID- 9744215 TI - Education and training in residential dementia care in Australia: needs, provision and directions. AB - This paper summarises existing education and training in dementia for aged care workers in Australia. The majority of aged care workers have no formal qualifications, while those with formal qualifications are mostly from a nursing background. Only half of nursing staff have attended any dementia care training. Existing training is either service based and provided in-house or by private consultants, or tertiary institution based and provided by academics and professional educators. There is considerable in-service and one-off service based training being provided around Australia, but few of these training exercises are linked to competency standards or staff appraisal. While there are some formal courses addressing training in dementia care available in every state of Australia, the emphasis on dementia care within generalist tertiary institution courses for aged care workers varies considerably. PMID- 9744216 TI - Outcome evaluation of an emergency department protocol of care on partner abuse. AB - STUDY OBJECTIVE: To evaluate the impact of a protocol on partner abuse (PA) at increasing identification and improving acute management of abused women by emergency department (ED) staff. METHODS: A community intervention trial compared two public hospital EDs at baseline and following implementation of a PA intervention. The intervention involved training staff at one ED in a protocol for the identification and acute management of abused women. Outcomes were assessed by reviewing a random sample of women's medical records. Identification of PA was assessed for each record on a yes/no basis. Identified cases were classified as 'confirmed' or 'suspected' PA. Acute management was assessed by ascertaining staff documentation of abuse and use of interventions. RESULTS: Approximately equal numbers of records were reviewed at each ED, pre and post implementation (total n = 8,051). Eighty-nine per cent of ED staff were trained. No difference in the overall identification of PA was found (chi 2 = 0.13, p = 0.72), but logistic regression analyses showed other significant changes. At the intervention site, there was an increase in confirmed cases of PA (chi 2 = 7.6, p = 0.006), a trend towards increased documentation (chi 2 = 3.5, p = 0.06) and a significant increase in interventions offered (chi 2 = 13.8, p = 0.002). Changes at the comparison site failed to reach significance. CONCLUSION: Implementation of this protocol resulted in a moderate increase in confirmed cases of abuse and improved the acute management offered to identified victims. The findings reinforce recommendations for widespread implementation of training and protocols to address partner abuse. PMID- 9744217 TI - Caesarean or vaginal birth: perceptions and experience of Thai women in Australian hospitals. AB - This paper discusses the perceptions and experience of immigrant Thai women who have had a Caesarean or vaginal birth. The ethnographic study showed that there was a considerably high number of emergency Caesarean births among Thai mothers. All women, except one, had prepared for a vaginal birth. However, most had to accept an emergency Caesarean and were told of the birth process just before the birth had taken place. Many women were, therefore, not well prepared for the birth. Most women preferred a vaginal birth over Caesarean birth. However, some believed that Caesarean was a safe method of birth because of the benefits of medical technology. This paper also examines some explanations for a high incidence of emergency Caesarean among Thai women in this study. PMID- 9744218 TI - Predictors of GP service use: a community survey of an elderly Australian sample. AB - As part of a study of an elderly community-dwelling Australian population, predictors of general practitioner (GP) service use were identified. The sample of 897 persons, aged 70 years or older and living in Canberra and Queanbeyan, were interviewed about their health and well-being. Data on the number of GP visits in the following 12-month period were obtained from the Health Insurance Commission. There were important gender differences in the prediction of both contact and volume of service use. Need variables (physical health in men, and disability and anxiety in women) were the most important predictors. Men who were older or who had lower occupational status used more medical services, as did women with less education or higher levels of extraversion. Men with lower social support were less likely to contact a GP, but social support was not related to volume of service use for either men or women. Since at most 21% of the variance in the volume of GP service use could be explained, despite the wide range of predictors considered and the different statistical approaches adopted, better measures of service use and predictors need to be developed. PMID- 9744219 TI - Caregiving and well-being in a sample of women in midlife. AB - A survey of 742 women in midlife found that 12% were involved in the care of another person with a chronic health disorder. Of note was the diversity of circumstances that led to the women becoming caregivers. Further, about a third of the carers were supporting more than one person. Carers did not differ on measures of depression or subjective health ratings from persons not involved in care. Burden scores were predicted by co-residence, low satisfaction with social support, and poorer health ratings on the part of the carers but not by the relationship between the person cared for and the carer. PMID- 9744220 TI - Health habits and risk of cognitive impairment and dementia in old age: a prospective study on the effects of exercise, smoking and alcohol consumption. AB - Previous research has yielded inconsistent results on the effects of exercise, smoking and alcohol use on cognitive impairment and dementia in old age. We analysed data from the Sydney Older Persons Study to see if these health habits were associated with cognitive functioning, dementia or Alzheimer's disease. Health habits were assessed in Wave 1 of the study, when the subjects were aged 75 years or over. Three years later, the subjects were tested for cognitive functioning and clinically examined for dementia and Alzheimer's disease. The analysis was restricted to the 327 subjects examined in Wave 2 who were non demented in Wave 1. There were few significant associations between health habits and cognitive performance and these were not found consistently across cognitive measures. No associations were found with dementia or Alzheimer's disease. While these health habits do not affect risk for dementia and cognitive impairment in the very elderly, who are at highest risk for these disorders, we cannot discount a role at younger ages. PMID- 9744221 TI - Estimating the cost of hospital treatment for injuries using linked morbidity data. AB - This paper describes the analysis of injury-related linked hospital morbidity data by admissions and by individual patients in Western Australia (WA) from 1990 to 1994. Over this five-year period, there were an average of 35,385 admissions and 30,524 people admitted each year for injuries in WA. The age-standardised rates for injury-related hospital admissions and persons admitted for injuries increased significantly, by 2.4% and 1.5% per year respectively, over the five year period. The number of admissions and the number of persons admitted peaked in the 20-24 years age group but the highest rates were among those aged 75 years and above. Injuries accounted for nearly 10% of all hospital bed day costs and cost about $50 per head of population per year. The cost of hospitalisation rose steadily from $85.2 million in 1990 to $113.6 million in 1994, the average cost being nearly $100 million per year. The average cost per injury related hospital episode was $2,748. Generally, the cost per hospital episode was higher for males and increased with age, following a similar pattern to that for the average length of stay. PMID- 9744222 TI - Defining breastfeeding. PMID- 9744223 TI - Access to cigarettes and adolescent smoking. PMID- 9744224 TI - Childhood vaccine argument questioned. PMID- 9744225 TI - Low lead levels in amniotic fluid and cord blood in a public hospital population. PMID- 9744226 TI - Ribena Tooth Kind. PMID- 9744227 TI - What on earth made you want to become a dentist? AB - The title, of course, came from a patient. The precise and original answer to the question has been largely overwhelmed by the vast changes in the profession over 30-odd years. The author discusses why he is glad he chose general dental practice; why it remains a career of interest, stimulus, challenge and satisfaction; and why he would commend it enthusiastically to today's school leavers. PMID- 9744228 TI - Scientific evidence to back up claims. PMID- 9744229 TI - Sentimentalising a potential health risk. PMID- 9744230 TI - Oral hygienists make a good impression. PMID- 9744231 TI - Clinical trials in primary dental care. PMID- 9744232 TI - Treatment refusal. PMID- 9744233 TI - Inappropriate prescription. PMID- 9744234 TI - An unusual case of trauma: a toothbrush embedded in the buccal mucosa. AB - Severe trauma from a toothbrush is unusual in childhood. This case report illustrates a case where a toothbrush was so embedded in the buccal soft tissues that it needed to be removed under a general anaesthetic. Other cases are outlined where life-threatening conditions rapidly arose following toothbrush trauma to the oro-pharynx, both patients having been initially discharged without treatment from a casualty department. Apparently minor oro-pharyngeal lacerations should be approached with caution when a penetrating injury from a toothbrush or any object has occurred. PMID- 9744235 TI - Connecting to the Internet. 2. PMID- 9744236 TI - Periapical cemental dysplasia: a case of misdiagnosis. AB - The correct diagnosis of pathological lesions of endodontic origin should allow for differentiation from those arising from other sources. A case of periapical cemental dysplasia (cementoma) is presented, whereby incorrect diagnosis resulted in not only inappropriate treatment, but an endodontic mishap and the superimposition of acute apical periodontitis in a previously disease-free site. This case report highlights the need for appropriate examination, simple special tests and diagnosis prior to management of lesions of questionable aetiology. PMID- 9744237 TI - How well are complete dentures copied? AB - OBJECTIVE: To assess how accurately the same set of complete dentures could be copied by 12 different laboratories. DESIGN: A prospective study. SETTINGS: Completed by commercial laboratories used by GDPs in England (1996). Results were analysed and controlled by a university department. SUBJECTS (MATERIALS) AND METHODS: The master set was constructed to include a midline diastema and a bilateral posterior crossbite. Twelve sets of silicone moulds of the master denture were prepared. GDPs selected commercial laboratories of their choice to receive the work. MAIN OUTCOME MEASURES: All stages of the work were compared with the master dentures by returning them to a specially adapted articulator. Analysis was completed using preset criteria. RESULTS: Only five of the returned copies were considered to have both good quality acrylic resin work and clear details of the teeth in wax. Only four cases reproduced the diastema. After processing, the occlusal vertical dimension had been increased in eight of the cases and the bilateral crossbite had been eliminated in all cases. CONCLUSIONS: None of the final copy dentures were considered an accurate copy of the master dentures. There is considerable scope for further training of dental technicians in this important technique. PMID- 9744239 TI - Prevalence of periradicular periodontitis associated with crowned teeth in an adult Scottish subpopulation. AB - OBJECTIVE: To examine the periradicular status of crowned teeth in an adult population in Scotland. DESIGN: Examination of full-mouth periapical radiographs from 319 consecutive adult patients (7596 teeth) attending Glasgow and Dundee Dental Hospitals for clinical examination. METHODS: The periradicular status of teeth with a crown present was assessed to determine the presence of a radiolucency which may indicate pulpal disease. RESULTS: 63.3% (n = 202) of patients had at least one tooth that was crowned. The total number of crowns assessed was 802, of which 458 (57.1%) were vital preparations, and 87 (19.0%) of these had radiographic signs of periradicular disease. The majority of the teeth (62.0%) had distinct widening of the periodontal membrane space which is considered to be an early sign of periapical disease. 42.9% (n = 344) of the crowned teeth had previous root canal treatment of which 50.8% (175) had evidence of a periradicular radiolucency. CONCLUSIONS: Pulpal damage may occur during procedures to provide a crown which may require subsequent root canal treatment. Radiographic follow-up of crowned teeth should be undertaken routinely. PMID- 9744240 TI - Issues concerning the development of a competency-based assessment system for dentistry. AB - The issue of standards of competence and assessment is central to professions. For the dental profession there have been suggestions that a change from overall competence to specific fields of competence would be a more suitable mechanism to assess whether an individual was competent. This paper examines the implications of such proposed changes and highlights several key issues, answers to which need to be provided prior to adopting new proposals. Failure to do so may well provide additional problems without addressing shortcomings in the present system. PMID- 9744238 TI - Anxiety levels, patient satisfaction and failed appointment rate in anxious patients referred by general practitioners to a dental hospital unit. AB - OBJECTIVE: To reduce the high failed appointment rate and anxiety levels among previously identified anxious new patients referred by general practitioners to a unit of restorative dentistry. A letter was drawn up which contained more explanatory information about the purpose and content of the initial dental appointment. DESIGN: The study was a single centre, double blind trial. SETTING: Referrals of nervous/anxious patients from general practitioners to the Manchester Dental Hospital. SUBJECTS: 185 patients were randomly allocated to the control group (n = 94) who received the standard hospital appointment information and the experimental group (n = 91) who received the new more informative letter. MAIN OUTCOME MEASURES: The effect of the new letter on patient attendance was recorded and anxiety levels were measured before and after seeing the clinician. RESULTS: 70 patients attended from the control group and 58 patients from the experimental group, giving 59 and 44 fully completed forms respectively. Both groups of patients were generally happy with the information that they were given. The attendance of patients in the experimental group and the control group were not significantly different. Preconsultation state anxiety levels were not reduced in the experimental group by the provision of the additional information but after the consultation both groups showed a significant reduction in state anxiety (P < 0.001). CONCLUSION: The new more informative letter did not improve attendance among this group of nervous patients. Other strategies for increasing initial attendance will therefore need to be identified and evaluated. In this study, the most important factor in reducing anxiety levels during the consultation appeared to be contact with the clinician rather than preparatory written postal information. PMID- 9744242 TI - Commentary: smallpox eradication in west and central Africa revisited. AB - In May 1980, the Thirty-third World Health Assembly adopted a resolution accepting the report of the Global Commission for the Certification of Smallpox Eradication and affirming its belief that this once-universal disease had been eradicated worldwide, 21 years after the global eradication programme had begun in 1959. A key element in the eradication effort was the surveillance-containment strategy, which was first tested in Nigeria in 1966, and which led to its adoption throughout the world. West and Central Africa became the first region of the world to be smallpox free and one by one other regions followed. One of the major lessons to be learned from the smallpox eradication programme is that interdependence is required if global results are to be achieved. Unfortunately, however, humanity has failed to learn this lesson in the long-term, and although global health has improved dramatically the gaps between the rich and poor remain vast. PMID- 9744243 TI - WHO Global Oral Data Bank, 1986-96: an overview of oral health surveys at 12 years of age. AB - The global oral health situation of 12-year-old children--decayed, missing, filled teeth (DMFT) index and the percentage of population affected--is described in this article using the latest representative studies for 80 countries included in the WHO Global Oral Data Bank (GODB) between 1986 and 1996. The quantity of information varied considerably: 68% of developed market economies had at least one national data set, compared with 38% of developing countries and 36% of economies in transition. By WHO region, the proportions were as follows: Eastern Mediterranean, 55%; European, 50%; Western Pacific, 48%; African, 39%; South-East Asia, 30%; and the Americas, 26%. Globally, the weighted DMFT index for all data in the GODB is < 3.0%, the WHO/Federation Dentaire Internationale goal for the year 2000. For the data reviewed in this article, achievement and nonachievement of this goal are discussed, as is the variation in DMFT means and proportions of children affected for various country groupings. There are difficulties in obtaining recent data for many countries, but the article emphasizes the need to maintain and develop the GODB to facilitate the compilation of valid, reliable and comparable data on oral health. PMID- 9744244 TI - Prevention and control of enterohaemorrhagic Escherichia coli (EHEC) infections: memorandum from a WHO meeting. WHO Consultation on Prevention and Control of Enterohaemorrhagic Escherichia coli (EHEC) Infections. AB - Escherichia coli is a commonly occurring inhabitant of the intestine of humans and other animals, but there are several pathogenic types of E. coli which cause a variety of human diseases. One of these pathogenic types, E. coli O157:H7, belongs to the group of enterohaemorrhagic E. coli (EHEC) which produce potent toxins and cause a particularly severe form of disease, haemorrhagic colitis (HC). About 10% of patients with HC can go on to develop haemolytic uraemic syndrome (HUS), a life-threatening complication of E. coli O157:H7 infection that is characterized by acute renal failure, haemolytic anaemia, and thrombocytopenia. These sequelae are particularly serious in young children and older people. On average, 2-7% of patients with HUS die, but in some outbreaks among the elderly the mortality rate has been as high as 50%. This Memorandum reviews the growing importance of E. coli O157:H7 as a foodborne pathogen and reports on the issues of surveillance, outbreak investigation, and control strategies with respect to EHEC infections that were discussed at the WHO Consultation on Prevention and Control of EHEC Infections, held in Geneva on 28 April to 1 May 1997. Recommended measures for prevention and control include the following: use of potable water in food production; presentation of clean animals at slaughter; improved hygiene throughout the slaughter process; appropriate use of food processing measures; thorough cooking of foods; and the education of food handlers, abattoir workers, and farm workers on the principles and application of food hygiene. PMID- 9744245 TI - Packaged treatment for first-line care in cerebral malaria and meningitis. AB - Described are the results of a trial carried out from January to June 1996 in southern Malawi to determine the effectiveness of a treatment pack for infants and children under the age of 6 years, who presented as emergencies to rural health centres with presumptive diagnoses of severe/cerebral malaria or meningitis. Each complete treatment pack (approximate cost, US$ 6) contained, inter alia, intramuscular quinine, intramuscular choloramphenicol, dextrose, paraldehyde, a nasogastric tube, prepacked syringes, and sterile water. A modified coma score and drug dosage nomogram were also included in the package. Despite a considerable drop in overall mortality, problems arose with regard to the incomplete treatment of possible meningitis and in the development of a rational referral policy. PMID- 9744246 TI - Resurgence of vivax malaria in Henan Province, China. AB - Henan Province (population, 90 million) in China has nonstable endemic malaria. After 1970 when 10.2 million cases of malaria were reported in the province, a huge control programme was undertaken, and in the mid-1980s indoor spraying and bednet impregnation with pyrethroids began. By 1992 only 318 cases were reported. In 1992 Henan declared "basic elimination of malaria" and in consequence spraying and bednet impregnation ceased after 1994. Subsequently, malaria broke out again in southern Henan. In 1995 we conducted a household survey for malaria transmission in southern Henan. Blood smears and serum samples for immunofluorescent antibody (IFA) testing were collected from 2329 people and 3.1% (73/2329) were positive for infection with Plasmodium vivax and 13% (301/2329) positive for malaria (titre > or = 1:20). All age groups were affected. Exophilic Anopheles sinensis occurs throughout the province; endo-anthropophilic A. anthropophagus, whose vectorial capacity is 20 times greater than that of A. sinensis, occurs mainly in southern Henan (S of latitude 33 degrees N) and was greatly reduced in numbers during 1985-92. Comparison of 1995 entomological data with historical data showed that A. anthropophagus increased in proportion to other anophelines after spraying activities and impregnation of bednets ceased. Over 10% of 9377 residents reported having malaria. The true number affected among the at-risk population of 700,000 must be larger. We conclude that impregnated bednets and malaria surveillance should continue even after an area is declared to have "basically eliminated" malaria. PMID- 9744247 TI - Prescribing patterns among medical practitioners in Pune, India. AB - In 1975 the World Health Assembly requested the Director-General to advise Member States on the selection and procurement of essential drugs corresponding to their national health needs. We report here the results of a study of the prescribing patterns and rational drug utilization of medical practitioners of Pune, an industrial city in the west of India, which was undertaken by analysing their prescriptions. The results indicated a lack of rational prescribing practices by a significant number of practitioners. Fixed-dose formulations dominated the prescribing pattern and generic prescriptions were negligible, with prescriptions for essential drugs accounting for less than 60% of the total number of drugs prescribed. More than 30% of prescriptions were irrational, with the probability of such prescriptions increasing significantly with the number of drugs per prescription. A study of sources of drug formulations available for prescription revealed significantly more fixed-dose combinations, many of which were irrational. These results call for intervention strategies to promote rational drug therapy in India. PMID- 9744248 TI - Reproductive tract infections, gynaecological morbidity and HIV seroprevalence among women in Mumbai, India. AB - Reported are the prevalence of reproductive tract infections and their contribution to pelvic inflammatory disease (PID), as well as the seroprevalence of human immunodeficiency virus (HIV), among women living in three inner city wards of Mumbai, India. Women aged < or = 35 years were recruited and screened as cases if they had been admitted to hospital for gynaecological investigation for suspected PID (n = 151) or infertility (n = 295); controls were healthy fertile women attending for laparoscopic tubal ligation (n = 2433). The women were mainly of low socioeconomic status. A total of 59.4% were migrants and 14.9% of these came to Mumbai to seek treatment. Cases reported a history of adverse pregnancy outcomes significantly more often than controls, and 30.5% of suspected PID cases had previously undergone laparoscopic tubal ligation. At examination 24.2% of cases and 8.4% of controls had a vaginal discharge. Pelvic infection was confirmed in 42.0% of suspected PID cases and 14.6% of infertile cases for whom diagnostic laparoscopy was performed. The prevalence of sexually transmitted diseases was low: Chlamydia trachomatis was found in 0.2%; and Neisseria gonorrhoeae was cultured from the cervix in only four cases. Neither of these infections was detected in laparoscopic aspirates. The prevalence of HIV1/2 infections in unlinked samples was 1.9%. Sexually transmitted diseases were not major factors leading to gynaecological morbidity. Heterosexual spread of HIV infection to this population of married women is still relatively low but needs to be carefully monitored. The gynaecological morbidity detected may be a consequence of widespread use of invasive methods of fertility regulation. PMID- 9744249 TI - Cutaneous leishmaniasis in primary school children in the south-eastern Iranian city of Bam, 1994-95. AB - Between August 1994 and July 1995, 11,517 primary school children aged 6-11 years in the south-eastern Iranian city of Bam, comprising 5560 (48.3%) girls and 5957 (51.7%) boys, were examined for the presence of active lesions or scars of cutaneous leishmaniasis (CL). There was a trend towards increasing prevalence with age, the prevalence being 10.7% in 6-year-old and 20% in > or = 11-year-old children. Overall, 1.3% of the children had active lesions and 14.3% had scars. There was no significant difference between the sexes in the prevalence of active lesions and/or scars. Of the children examined, 54 (0.5%) had leishmaniasis recidivans: 19 girls (35.2%) and 35 boys (64.8%). The number of active lesions or scars per child ranged from 1 to 10. The majority (82.3%) had 1 lesion, 12.4% had 2 lesions, and 5.3% had > or = 3. The average number of lesions was 1.08 (1.03 in girls and 1.18 in boys). The face was the part of the body most commonly involved (63.6%), followed by the hands (20.9%), legs (12.8%) and other parts of the body (2.7%). Examination of isolates from 14 children revealed that in 13 (92.9%) the causal organism was Leishmania tropica and in the other (7.1%) L. major. The survey indicates that the geographical distribution of CL is far wider than previously thought. It also shows that Bam is a suitable areas for a vaccine field trial. PMID- 9744250 TI - Measurement and determinants of tuberculosis outcome in Karonga District, Malawi. AB - Evaluation of disease outcome is central to the assessment of tuberculosis (TB) control programmes. In the study reported in this article we examined the factors influencing the measurement of outcome, survival rates during and after treatment, smear conversion rates, and relapse rates for patients diagnosed with TB in a rural area of Malawi between 1986 and mid-1994. Patients with less certain diagnoses of TB were more likely to die than those with confirmed TB, both among those who were seropositive and those who were seronegative to human immunodeficiency virus (HIV). The mortality rate among smear-positive patients with a separate culture-positive specimen was half that of patients with no such diagnostic confirmation. Patients not registered by the Ministry of Health had much higher mortality and default rates than did registered patients. Among smear positive patients, HIV serostatus was the most important influence on mortality both during and after treatment (crude hazard ratios (95% confidence intervals) = 5.6 (3.0-10) and 7.7 (3.4-17), resp.), but HIV serostatus did not influence smear conversion rates. The initial degree of smear positivity influenced smear conversion rates, but not mortality rates. No significant predictors of relapse were identified. Unless considerable care is taken to include all TB patients, and to exclude nontuberculous patients, recorded TB outcome statistics are difficult to interpret and may be misleading. In populations with high rates of HIV infection, TB target cure rates of 85% are unrealistic. When new interventions are assessed it cannot be assumed that factors which influence the smear conversion rate will also influence the mortality rate. PMID- 9744251 TI - Evaluation and determinants of outcome of tuberculosis treatment. AB - The most important determinant of the outcome of treatment of tuberculosis (TB) patients is the access that they have to reliable diagnosis and treatment services organized within a national tuberculosis programme (NTP) as part of DOTS, the name of the WHO-recommended TB control strategy. Accurate evaluation of treatment outcomes requires inclusion of all patients diagnosed, including those not registered; part of routine NTP management should therefore involve regular cross-checks between laboratory diagnostic registers and NTP treatment registers. Finally, control services should take into consideration the type of TB, the presence of concomitant infection with human immunodeficiency virus (HIV), and patient characteristics. PMID- 9744252 TI - Brief communication: rapid culture of tubercle bacilli. AB - One of the biggest obstacles to the correct diagnosis and efficient treatment of tuberculosis is the absence of a rapid technique for culturing tubercle bacilli and for testing their susceptibility to antituberculosis drugs. Current procedures typically take 6-10 weeks to perform. This article describes a simple, rapid, reliable and cheap method of culturing tubercle bacilli using a liquid medium consisting of a mixture of coconut water, horse serum, glycerol and benzylpenicillin. Addition of specific concentrations of antituberculosis drugs to the medium, permits information on the drug susceptibility of tubercle bacilli to be obtained in only 6 days. The procedure requires no special instruments or technical skill and can therefore be carried out routinely in the average laboratory in developing countries. PMID- 9744253 TI - Cord blood banking. PMID- 9744254 TI - Young scientists deserve better of the system (yet again) PMID- 9744255 TI - Jobs crisis sparks call for freeze in number of PhD students in US. PMID- 9744256 TI - Retraining scheme could go Europe-wide. PMID- 9744257 TI - Stock market crash threatens start-up biotech companies. PMID- 9744258 TI - Call for Europe-wide public health agency. PMID- 9744259 TI - Rapid shake-up at German health institute. PMID- 9744260 TI - US unlikely to swallow plan for a food safety supremo. PMID- 9744261 TI - Row in India over rules on animal experiments. PMID- 9744262 TI - Controversial appointment to SA council. PMID- 9744263 TI - Casting an eye over cyclopia. PMID- 9744264 TI - Ancient cosmic spherules. PMID- 9744265 TI - HIV. Setting death in motion. PMID- 9744266 TI - Perceptual bias for rising tones. PMID- 9744267 TI - p14ARF links the tumour suppressors RB and p53. PMID- 9744268 TI - p19ARF links the tumour suppressor p53 to Ras. PMID- 9744269 TI - On-line journals and financial fire walls. PMID- 9744270 TI - Ligand binding and co-activator assembly of the peroxisome proliferator-activated receptor-gamma. AB - The peroxisome proliferator-activated receptor-gamma (PPAR-gamma) is a ligand dependent transcription factor that is important in adipocyte differentiation and glucose homeostasis and which depends on interactions with co-activators, including steroid receptor co-activating factor-1 (SRC-1). Here we present the X ray crystal structure of the human apo-PPAR-gamma ligand-binding domain (LBD), at 2.2 A resolution; this structure reveals a large binding pocket, which may explain the diversity of ligands for PPAR-gamma. We also describe the ternary complex containing the PPAR-gamma LBD, the antidiabetic ligand rosiglitazone (BRL49653), and 88 amino acids of human SRC-1 at 2.3 A resolution. Glutamate and lysine residues that are highly conserved in LBDs of nuclear receptors form a 'charge clamp' that contacts backbone atoms of the LXXLL helices of SRC-1. These results, together with the observation that two consecutive LXXLL motifs of SRC-1 make identical contacts with both subunits of a PPAR-gamma homodimer, suggest a general mechanism for the assembly of nuclear receptors with co-activators. PMID- 9744271 TI - A mysterious dust clump in a disk around an evolved binary star system. AB - The discovery of planets in orbit around the pulsar PSR1257+12 shows that planets may form around post-main-sequence stars. Other evolved stars, such as HD44179 (an evolved star which is part of the binary system that has expelled the gas and dust that make the Red Rectangle nebula), possess gravitationally bound orbiting dust disks. It is possible that planets might form from gravitational collapse in such disks. Here we report high-angular-resolution observations at millimetre and submillimetre wavelengths of the dusk disk associated with the Red Rectangle. We find a dust clump with an estimated mass near that of Jupiter in the outer region of the disk. The clump is larger than our Solar System, and far beyond where planet formation would normally be expected, so its nature is at present unclear. PMID- 9744272 TI - Unaltered cosmic spherules in a 1.4-Gyr-old sandstone from Finland. AB - Micrometeorites-submillimetre-sized particles derived from asteroids and comets occur in significant quantities in deep sea sediments, and the ice sheets of Greenland and Antarctica. The most abundant micrometeorites are cosmic spherules, which contain nickel-rich spinels that were crystallized and oxidized during atmospheric entry, therefore recording the oxygen content in the uppermost atmosphere. But the use of micrometeorites for detecting past changes in the flux of incoming extraterrestrial matter, and as probes of the evolution of the atmosphere, has been hampered by the fact that most objects with depositional ages higher than 0.5 Myr show severe chemical alteration. Here we report the discovery of unaltered cosmic spherules in a 1.4-Gyr-old sandstone (red bed) from Finland. From this we infer that red beds, a common lithology in the Earth's history, may contain substantial unbiased populations of fossil micrometeorites. The study of such populations would allow systematic research on variations in the micrometeorite flux from the early Proterozoic era to recent times (a time span of about 2.5 Gyr), and could help to better constrain the time when the atmospheric oxygen content was raised to its present level. PMID- 9744273 TI - Prism adaptation to a rightward optical deviation rehabilitates left hemispatial neglect. AB - A large proportion of right-hemisphere stroke patients show hemispatial neglect-a neurological deficit of perception, attention, representation, and/or performing actions within their left-sided space, inducing many functional debilitating effects on everyday life, and responsible for poor functional recovery and ability to benefit from treatment. The frequent parietal locus of the lesion producing neglect reflects the impairment of coordinate transformation used by the nervous system to represent extrapersonal space. Given that adaptation to a visual distortion can provide an efficient way to stimulate neural structures responsible for the transformation of sensorimotor coordinates, the aim of our study was to investigate the effect of prism adaptation on various neglect symptoms, including the pathological shift of the subjective midline to the right. All patients exposed to the optical shift of the visual field to the right were improved on their manual body-midline demonstration and on classical neuropsychological tests. Unlike other physiological manipulations used to improve neglect, this improvement lasted for at least two hours after prism removal and thus could be useful in rehabilitation programmes. The positive effect found for both sensorimotor and more cognitive spatial functions suggests that they share or depend on a common level of space representation linked to multisensory integration. PMID- 9744274 TI - Phasic alerting of neglect patients overcomes their spatial deficit in visual awareness. AB - Patients with extensive damage to the right hemisphere of their brain often exhibit unilateral neglect of the left side of space. The spatial attention of these patients is strongly biased towards the right, so their awareness of visual events on the left is impaired. Extensive right-hemisphere lesions also impair tonic alertness (the ability to maintain arousal). This nonspatial deficit in alertness is often considered to be a different problem from spatial neglect, but the two impairments may be linked. If so, then phasically increasing the patients' alertness should temporarily ameliorate their spatial bias in awareness. Here we provide evidence to support this theory. Right-hemisphere neglect patients judged whether a visual event on the left preceded or followed a comparable event on the right. They became aware of left events half a second later than right events on average. This spatial imbalance in the time course of visual awareness was corrected when a warning sound alerted the patients phasically. Even a warning sound on the right accelerated the perception of left visual events in this way. Nonspatial phasic alerting can thus overcome disabling spatial biases in perceptual awareness after brain injury. PMID- 9744275 TI - Increased number of synaptic GABA(A) receptors underlies potentiation at hippocampal inhibitory synapses. AB - Changes in synaptic efficacy are essential for neuronal development, learning and memory formation and for pathological states of neuronal excitability, including temporal-lobe epilepsy. At synapses, where there is a high probability of opening of postsynaptic receptors, all of which are occupied by the released transmitter, the most effective means of augmenting postsynaptic responses is to increase the number of receptors. Here we combine quantal analysis of evoked inhibitory postsynaptic currents with quantitative immunogold localization of synaptic GABA(A) receptors in hippocampal granule cells in order to clarify the basis of inhibitory synaptic plasticity induced by an experimental model of temporal-lobe epilepsy (a process known as kindling). We find that the larger amplitude (66% increase) of elementary synaptic currents (quantal size) after kindling results directly from a 75% increase in the number of GABA(A) receptors at inhibitory synapses on somata and axon initial segments. Receptor density was up by 34-40% and the synaptic junctional area was expanded by 31%. Presynaptic boutons were enlarged, which may account for the 39% decrease in the average number of released transmitter packets (quantal content). Our findings establish the postsynaptic insertion of new GABA(A) receptors and the corresponding increase in postsynaptic responses augmenting the efficacy of mammalian inhibitory synapses. PMID- 9744276 TI - Cloning of inv, a gene that controls left/right asymmetry and kidney development. AB - Most vertebrate internal organs show a distinctive left/right asymmetry. The inv (inversion of embryonic turning) mutation in mice was created previously by random insertional mutagenesis; it produces both a constant reversal of left/right polarity (situs inversus) and cyst formation in the kidneys. Asymmetric expression patterns of the genes nodal and lefty are reversed in the inv mutant, indicating that inv may act early in left/right determination. Here we identify a new gene located at the inv locus. The encoded protein contains 15 consecutive repeats of an Ank/Swi6 motif at its amino terminus. Expression of the gene is the highest in the kidneys and liver among adult tissues, and is seen in presomite-stage embryos. Analysis of the transgenic genome and the structure of the candidate gene indicate that the candidate gene is the only gene that is disrupted in inv mutants. Transgenic introduction of a minigene encoding the candidate protein restores normal left/right asymmetry and kidney development in the inv mutant, confirming the identity of the candidate gene. PMID- 9744277 TI - Zebrafish organizer development and germ-layer formation require nodal-related signals. AB - The vertebrate body plan is established during gastrulation, when cells move inwards to form the mesodermal and endodermal germ layers. Signals from a region of dorsal mesoderm, which is termed the organizer, pattern the body axis by specifying the fates of neighbouring cells. The organizer is itself induced by earlier signals. Although members of the transforming growth factor-beta (TGF beta) and Wnt families have been implicated in the formation of the organizer, no endogenous signalling molecule is known to be required for this process. Here we report that the zebrafish squint (sqt) and cyclops (cyc) genes have essential, although partly redundant, functions in organizer development and also in the formation of mesoderm and endoderm. We show that the sqt gene encodes a member of the TGF-beta superfamily that is related to mouse nodal. cyc encodes another nodal-related proteins, which is consistent with our genetic evidence that sqt and cyc have overlapping functions. The sqt gene is expressed in a dorsal region of the blastula that includes the extraembryonic yolk syncytial layer (YSL). The YSL has been implicated as a source of signals that induce organizer development and mesendoderm formation. Misexpression of sqt RNA within the embryo or specifically in the YSL induces expanded or ectopic dorsal mesoderm. These results establish an essential role for nodal-related signals in organizer development and mesendoderm formation. PMID- 9744278 TI - Induction of the zebrafish ventral brain and floorplate requires cyclops/nodal signalling. AB - Zebrafish cyclops (cyc) mutations cause deficiencies in the dorsal mesendoderm and ventral neural tube, leading to neural defects and cyclopia. Here we report that cyc encodes a transforming growth factor-beta (TGF-beta)-related intercellular signalling molecule that is similar to mouse nodal. cyc is expressed in dorsal mesendoderm at gastrulation and in the prechordal plate until early somitogenesis. Expression reappears transiently in the left lateral-plate mesoderm, and in an unprecedented asymmetric pattern in the left forebrain. Injection of cyc RNA non-autonomously restores sonic hedgehog-expressing cells of the ventral brain and floorplate that are absent in cyc mutants, whereas inducing activities are abolished by cyc, a mutation of a conserved cysteine in the mature ligand. Our results indicate that cyc provides an essential non-cell-autonomous signal at gastrulation, leading to induction of the floorplate and ventral brain. PMID- 9744279 TI - Apoptosis of CD8+ T cells is mediated by macrophages through interaction of HIV gp120 with chemokine receptor CXCR4. AB - CD8-positive T cells are thought to play an important role in the control of infection by human immunodeficiency virus (HIV) as a result of their cytotoxic activity and by releasing soluble factors. In AIDS patients, the absolute number of CD8+ T lymphocytes is decreased in peripheral blood and their turnover rate is increased, suggesting that there is more cell renewal and cell death occurring. Anti-retroviral therapy raises CD8+ T-cell counts in HIV-infected patients. Here we report that the death rate of CD8+ T cells by apoptosis increased markedly during HIV infection of peripheral blood mononuclear cells in vitro. Apoptosis is induced in a dose-dependent manner by recombinant envelope glycoprotein gp120 from HIV strain X4, or by stromal-derived factor-1 (SDF-1), the physiological ligand of the chemokine receptor CXCR4. Apoptosis is mediated by the interaction between tumour-necrosis factor-alpha bound to the membrane of macrophages (mbTNF) and a receptor on CD8+ T cells (TNF-receptor II, or TNFRII). The expression of both of these cell-surface proteins is upregulated by HIV infection or by treatment with recombinant gp120 or SDF-1. Apoptosis of CD8+ T cells isolated from HIV-infected patients is also mediated by macrophages through the interaction between mbTNF and TNFRII. These results indicate that the increased turnover of CD8+ T cells in HIV-infected subjects is mediated by the HIV envelope protein through the CXCR4 chemokine receptor. PMID- 9744280 TI - The p35/Cdk5 kinase is a neuron-specific Rac effector that inhibits Pak1 activity. AB - Cyclin-dependent kinase 5 (Cdk5) and its neuron-specific regulator p35 are essential for neuronal migration and for the laminar configuration of the cerebral cortex. In addition, p35/Cdk5 kinase concentrates at the leading edges of axonal growth cones and regulates neurite outgrowth in cortical neurons in culture. The Rho family of small GTPases is implicated in a range of cellular functions, including cell migration and neurite outgrowth. Here we show that the p35/Cdk5 kinase co-localizes with Rac in neuronal growth cones. Furthermore, p35 associates directly with Rac in a GTP-dependent manner. Another Rac effector, Pak1 kinase, is also present in the Rac-p35/Cdk5 complexes and co-localizes with p35/Cdk5 and Rac at neuronal peripheries. The active p35/Cdk5 kinase causes Pak1 hyperphosphorylation in a Rac-dependent manner, which results in down-regulation of Pak1 kinase activity. Because the Rho family of GTPases and the Pak kinases are implicated in actin polymerization, the modification of Pak1, imposed by the p35/Cdk5 kinase, is likely to have an impact on the dynamics of the reorganization of the actin cytoskeleton in neurons, thus promoting neuronal migration and neurite outgrowth. PMID- 9744281 TI - Interactions controlling the assembly of nuclear-receptor heterodimers and co activators. AB - Retinoic-acid receptor-alpha (RAR-alpha) and peroxisome proliferator-activated receptor-gamma (PPAR-gamma) are members of the nuclear-receptor superfamily that bind to DNA as heterodimers with retinoid-X receptors (RXRs). PPAR-RXR heterodimers can be activated by PPAR or RXR ligands, whereas RAR-RXR heterodimers are selectively activated by RAR ligands only, because of allosteric inhibition of the binding of ligands to RXR by RAR. However, RXR ligands can potentiate the transcriptional effects of RAR ligands in cells. Transcriptional activation by nuclear receptors requires a carboxy-terminal helical region, termed activation function-2 (AF-2), that forms part of the ligand-binding pocket and undergoes a conformational change required for the recruitment of co activator proteins, including NCoA-1/SRC-1. Here we show that allosteric inhibition of RXR results from a rotation of the RXR AF-2 helix that places it in contact with the RAR coactivator-binding site. Recruitment of an LXXLL motif of SRC-1 to RAR in response to ligand displaces the RXR AF-2 domain, allowing RXR ligands to bind and promote the binding of a second LXXLL motif from the same SRC 1 molecule. These results may partly explain the different responses of nuclear receptor heterodimers to RXR-specific ligands. PMID- 9744282 TI - Targeting the subthalamic nucleus in the treatment of Parkinson's disease. AB - As more is learnt about the functional implications of basal ganglia connectivity, the role of the subthalamic nucleus as a target site for stereotactic procedures in the amelioration of the symptoms of Parkinson's disease is becoming clearer. A comparison of various neurosurgical procedures in the disease is discussed in relation to current thinking about circuitry. Experimental investigations involving lesioning or stimulation of the subthalamic nucleus in nonhuman primate models and in clinical studies of Parkinson's disease are compared. Neurosurgical procedures that lesion structures bilaterally are more likely to induce side effects than is deep-brain stimulation, which has the added advantage of reversibility and which is more amenable to titration in relation to medication and dosage. A small but growing number of parkinsonian patients have received subthalamic stimulation either unilaterally or bilaterally. Stimulation of the subthalamic nucleus ameliorates tremor, rigidity and hypokinesia, as opposed to thalamic stimulation which is probably best reserved for tremor-dominant patients. Such procedures also do not involve the same complex technical and ethical issues that are associated with foetal mesencephalic grafting. Although subthalamic stimulation shows great promise, it has not been developed to the point where it can be used as more than an experimental treatment. Further experimental research is required before the new strategies can be used on a larger scale. PMID- 9744283 TI - Cholinergic mechanism and blood pressure regulation in the central nervous system. AB - Cholinergic neurons in numerous brain regions have been implicated in blood pressure regulation. One of the most important brain regions where cholinergic neurons play a role in the pathogenesis of hypertension is the rostral ventrolateral medulla (RVL), an essential source of efferent sympathetic activity. Pharmacological and biochemical studies have revealed that acetylcholine release in the RVL is increased in experimental hypertension regardless of its etiology and that this enhanced release of acetylcholine leads to hypertension. The lateral parabrachial nucleus, another important hindbrain area involved in blood pressure regulation, is responsible for the enhanced release of acetylcholine in the RVL of hypertensive animals. Moreover, recent studies have demonstrated the involvement of the hypothalamic defence area, an area believed to be involved in the hypertension induced by chronic stress, in the release of acetylcholine in the RVL and also have demonstrated the existence of direct projections from the hypothalamic structures to the lateral parabrachial nucleus. More studies about mechanisms of the enhanced release of acetylcholine in the RVL of experimentally hypertensive animals will provide important information for central mechanisms of hypertension. PMID- 9744284 TI - Effect of glutamate receptor antagonists on excitatory postsynaptic potentials in striatum. AB - Glutamate, as the main transmitter of corticostriatal pathway, has a crucial role in the regulation of the activity of striatal cells as well as in pathogenesis of some diseases characterized by striatal malfunction caused by overexcitation of neurons. In the present study, the role of ionotropic excitatory amino acid receptors was investigated in the striatal synaptic transmission. Using conventional intracellular electrophysiological methods in brain slices, we have investigated the effects of the N-methyl-D-aspartate (NMDA) antagonist (+/-) 2 amino-5-phosphono-valerate (APV) and the alpha-amino-3-hydroxy-5-methyl-isoxazole 4-propionate (AMPA) antagonist (+/-) 1-(4-aminophenyl)-3-methyl-carbamoyl-7,8 methylenedioxy-5H-2,3-benzodiaz epine (GYKI 53655) on the excitatory postsynaptic potentials (EPSPs) evoked by electrical stimulation of corpus callosalpham. The AMPA antagonist significantly decreased electrically evoked responses and a weak inhibition was also observed after APV application. The results were compared to similar data obtained in a cortical slice study. PMID- 9744285 TI - Striatal lesions produce distinctive impairments in reaction time performance in two different operant chambers. AB - The dorsal striatum plays a crucial role in mediating voluntary movement. Excitotoxic striatal lesions in rats have previously been shown to impair the initiation but not the execution of movement in a choice reaction time task in an automated lateralised nose-poke apparatus (the "nine-hole box"). Conversely, when a conceptually similar reaction time task has been applied in a conventional operant chamber (or "Skinner box"), striatal lesions have been seen to impair the execution rather than the initiation of the lateralised movement. The present study was undertaken to compare directly these two results by training the same group of rats to perform a choice reaction time task in the two chambers and then comparing the effects of a unilateral excitotoxic striatal lesion in both chambers in parallel. Particular attention was paid to adopting similar parameters and contingencies in the control of the task in the two test chambers. After striatal lesions, the rats showed predominantly contralateral impairments in both tasks. However, they showed a deficit in reaction time in the nine-hole box but an apparent deficit in response execution in the Skinner box. This finding confirms the previous studies and indicates that differences in outcome are not simply attributable to procedural differences in the lesions, training conditions or tasks parameters. Rather, the pattern of reaction time deficit after striatal lesions depends critically on the apparatus used and the precise response requirements for each task. PMID- 9744286 TI - Area postrema removal abolishes stimulatory effects of intravenous interleukin 1beta on hypothalamic-pituitary-adrenal axis activity and c-fos mRNA in the hypothalamic paraventricular nucleus. AB - This study examined the role of the area postrema (AP) in transducing peripheral immune signals, represented by intravenous (i.v.) interleukin-1beta (IL-1), into neuroendocrine responses. The AP, a circumventricular organ with a leaky blood brain barrier, lies adjacent to the nucleus of the solitary tract (NTS) in the medulla. The AP was removed by aspiration, and 2 weeks later, AP-lesioned or sham lesioned rats were injected i.v. with 0.5 microg/kg IL-1 or sterile saline. After 30 min, brains were removed and analyzed for c-fos mRNA levels in various structures implicated in the hypothalamic-pituitary-adrenal axis response to peripheral cytokine challenge. The sham-lesioned animals responded to IL-1 with large elevations in adrenocorticotropic hormone (ACTH) and corticosterone levels in the plasma and c-fos mRNA levels in cells of the AP, NTS, central nucleus of the amygdala, bed nucleus of the stria terminalis, hypothalamic paraventricular nucleus (PVN), and meninges. Prior AP removal abolished the IL-1 -induced increases in ACTH and corticosterone in the plasma and c-fos mRNA levels in the NTS and PVN. However, AP removal had no effect on IL-1-induced increases in c-fos mRNA levels in the other areas examined. The selective AP lesion effects suggest that the AP and adjacent NTS play a pivotal role in transducing a circulating IL 1 signal into hypothalamic-pituitary-adrenal axis activation by a pathway that may be comprised of known anatomical links between the AP, NTS, and corticotropin releasing hormone neurons of the PVN. PMID- 9744287 TI - Metabotropic glutamate receptors are involved in calcium-induced LTP of AMPA and NMDA receptor-mediated responses in the rat hippocampus. AB - Effects of metabotropic glutamate (mGlu) receptors on calcium-induced long-term potentiation (LTP) of alpha-amino-3-hydroxy-5-methyl-4-isoxazoleproprionate (AMPA) and N-methyl-D-aspartate (NMDA) receptor-mediated components were investigated in rat hippocampal slices using whole-cell patch-clamp recordings of excitatory postsynaptic currents (EPSCs). Calcium-induced LTP comprises a parallel, long-lasting increase of AMPA and NMDA receptor-mediated components. The calcium-induced LTP of the AMPA receptor-mediated component can be significantly attenuated by the use of a selective NMDA antagonist. (R.S)-alpha methyl-4-carboxyphenylglycine (MCPG), a selective antagonist of mGlu receptors, abolished the long-lasting increase of both AMPA and NMDA receptor-mediated components observed in calcium-induced LTP. In current clamp mode, the application of a high calcium alone or Schaffer fiber stimulation alone (20 Hz) only generated a short-term increase in the firing rate of evoked action potentials. Conversely, a long-term increase in the firing rate was observed if Schaffer fiber stimulation (20 Hz) accompanied the perfusion of high calcium. These results suggest that calcium-induced LTP involves a parallel, long-lasting enhancement in ionotropic AMPA and NMDA receptor-mediated components. More importantly, the mGlu receptor plays a critical role in the establishment of both AMPA and NMDA receptor-mediated components underlying calcium-induced LTP. In addition, the present study also described an experimental condition in which the coapplication of the high calcium pulse and Schaffer fiber stimulation (20 Hz) can synergistically elicit a long-term increase of neuronal excitability. PMID- 9744288 TI - Effect of streptozotocin-induced diabetes on rat brain sulfonylurea binding sites. AB - Both high and low affinity sulfonylurea receptors (SURs) reside on glucose responsive neurons where they influence cell firing and neurotransmitter release via the adenosinetriphosphate (ATP)-sensitive K+ (katp) channel. Here, the effect of diabetes on [3H] glyburide binding to SURs was assessed in male obesity resistant Sprague-Dawley rats rendered diabetic with streptozotocin (65 mg/kg, i.p.). Additional streptozotocin-treated rats were supplemented with insulin (1.5 U/kg/ day). Streptozotocin reduced plasma insulin to 13% of control associated with hyperglycemia (25.3 +/- 1.7 mmol/l), while insulin lowered plasma glucose (9.56 +/- 1.78 mmol/l) to near control levels (7.65 +/- 0.22 mmol/l). Over 7 days, all streptozotocin-treated rats lost 12% of their initial body wt. while controls gained 1%. Despite equivalent wt. loss, streptozotocin-induced diabetes selectively increased high affinity [3H] glyburide binding in the hypothalamic dorsomedial nuclei (DMN) and ventromedial nuclei (VMN) and lateral area (LH). This was prevented by insulin injections. Low affinity binding was similarly increased in the DMN and VMN, as well as two amygdalar subnuclei but decreased in the substantia nigra, pars compacta. Insulin fully prevented these changes only in the DMN and one amygdalar nucleus and the substantia nigra. Therefore, binding to (SURs) appears to be generally upregulated in the face of hypoinsulinemia with hyperglycemia and this is prevented by insulin treatment. These and other data suggest that this combination of abnormalities in diabetes should have an adverse effect on the glucose sensing capacity of the brain. PMID- 9744289 TI - Morphological and functional changes after benzalkonium chloride treatment of the small intestinal Thiry-Vella loop in rats. AB - The aim of this work was to study the effects of benzalkonium chloride (BAC) treatment on the small intestine and its functioning in rats surgically prepared with Thiry-Vella intestinal loop. The loops were treated with either BAC, which ablated much of the myenteric plexus and extrinsic innervation, or with physiological saline (SAL). In vivo drinking experiments were performed to examine the effect on fluid intake and behavioral indices of distending the loop with a balloon. Spontaneous motility and its changes induced by acetylcholine (ACh) and histamine (His) were studied on isolated stripes in vitro. Finally, samples from the loops were examined histologically. Though reduction of the cell number was less than expected and no differences of the thickness of the muscular layer between the two groups was observed, BAC treatment altered the pattern of spontaneous activity and also the sensitivity to ACh and His in isolated stripes. In vivo distension of the SAL-treated loops reduced fluid intake and produced signs of aversivity; these effects were absent in the BAC-treated group. Our results show that despite the differences in the degree of ablation from those obtained by others, BAC treatment can be used to study the mechanisms underlying the effects of the enteral stimuli on the behavior. PMID- 9744290 TI - Zucker obese rats are insensitive to the CRH-increasing effect of oleoyl-estrone. AB - Adult female Zucker lean and obese rats were treated for 14 days with 3.5 nm/kg oleoyl-estrone (OE) in liposomes (Merlin-2) through continuous i.v. injection with osmotic minipumps. Rat wt. and food intake were measured daily. On days 0, 3, 6, 10, and 14, groups of rats were killed and their hypothalamic nuclei [lateral preoptic (LPO), median preoptic (MPO), paraventricular (PVN), ventromedial (VMH), and arcuate (ARC)] were dissected, homogenized, and used for the measurement of corticosterone-releasing hormone (CRH) by radioimmunoassay. The OE treatment decreased food intake by 67.4% in lean and 62.6% in obese rats (means for 14 days). Body wt. decreased steadily in lean and obese rats, the gap between controls and treated rats becoming 11.5% of initial body wt. in the lean and 12.4% in the obese. The levels of CRH in the ARC nucleus were at least 10 fold higher than in the other nuclei. No changes in CRH were observed in any of the nuclei of obese rats, with levels up to day 6 similar to those of lean rats. In the lean rats, the LPO and ARC nuclei showed peaks on day 10, while the MPO showed no changes and the PVN and VMH nuclei showed a progressive increase, to a maximum at the end of the study (day 14). This contrasted with the peak of plasma adrenocorticotropic hormone (ACTH) and corticosterone (day 6 in lean and day 14 in obese rats). There was a definite lack of correlation between the plasma levels of these two hormones and the levels of CRH in the hypothalamic nuclei, and between the latter and the decreases in appetite in the rats. The loss of appetite induced by OE is not necessarily mediated by CRH, because the obese rats show an intense decrease in voluntary food intake but their hypothalamic nuclei CRH levels do not change at all. Hypothalamic nuclei CRH does not, necessarily, mediate the rise in glucocorticoids induced by OE treatment, because this is observed in lean and obese rats, lean rats increases being mismatched with those of hypothalamic CRH. The OE induced changes in hypothalamic CRH require a fully functional leptinergic pathway, because it is not observed in Zucker fa/fa rats lacking a working leptin receptor. This--indirectly--shows that leptin is needed for its synthesis or modulation. PMID- 9744291 TI - Anti-convulsant and adverse effects of the glycineB receptor ligands, D cycloserine and L-701,324: comparison with competitive and non-competitive N methyl-D-aspartate receptor antagonists. AB - In this study, the anticonvulsant and adverse effects of compounds that belong to four different categories of systemically available N-methyl-D-aspartate (NMDA) receptor ligands were compared, namely the competitive antagonist CGP 40116, the noncompetitive antagonist MK-801 (dizocilpine), the glycineB receptor antagonist L-701,324, and the glycineB receptor high-efficacy partial agonist D-cycloserine. The maximal electroshock seizures (MES), which are widely used to detect drug efficacy against generalized tonic-clonic seizures in humans, were produced by transcorneal electrical stimulation. Abolition of tonic hind-limb extension was taken as the end-point. The drug-induced motor and long-term memory deficits were quantified by using the inverted screen test and the step-through passive avoidance test, respectively. All tested compounds exhibited significant anticonvulsant effect. The rank order of potency for the respective compounds was: MK-801 = CGP 40116 > L-701,324 >> D-cycloserine. All of these compounds induced motor impairment at doses close to those found to be anticonvulsant, however, hyperlocomotion and stereotyped behavior occurred only with MK-801. The highest protective indices [PI = TD50 (inverted screen)/ED50 (MES)] were calculated for CGP 40116 and D-cycloserine (2.4 and 2.2, respectively). When tested for memory impairment at one-half the MES ED50, again only MK-801 induced significant memory disruption in the passive avoidance test. In conclusion, these results suggest that glycineB receptor high-efficacy partial agonists and competitive NMDA receptor antagonists may be advantageous to noncompetitive NMDA antagonists and glycineB receptor antagonists as potential antiepileptic drugs. PMID- 9744292 TI - Nitric oxide and prostaglandin E2 formation parallels blood-brain barrier disruption in an experimental rat model of bacterial meningitis. AB - During meningitis, the host produces a plethora of signaling agents as part of a coordinated defense mechanism against invading pathogens. Nitric oxide (NO) and prostaglandin E2 (PGE2) are two such inflammatory mediators produced in response to bacterial endotoxins. Disruption of the blood-brain barrier (BBB) is one of many pathophysiological consequences of meningitis. The present objective was to examine the time-course of NO and PGE2 production in relationship to BBB permeability alterations during experimentally-induced meningitis. Meningeal inflammation was elicited by intracisternal administration of the bacterial endotoxin, lipopolysaccharides (LPS; 200 microg), and NO, PGE2, and BBB integrity were monitored over the next 24 h. Meningeal NO production was assessed by headspace chemiluminescence; cerebrospinal fluid PGE2 was determined by enzyme linked immunosorbent assay (ELISA) immunoassay; and BBB integrity was determined by the brain accumulation of 14C-sucrose. Similar time-course profiles for NO and PGE2 were observed, with a peak effect for both inflammatory mediators observed within 6-8 h after intracisternal LPS dosing. Statistically significant (p < 0.05) disruption of the BBB was observed in various brain regions. Strikingly similar temporal relationships were observed for NO and PGE2 production and BBB disruption. These results suggest the hypothesis that NO and PGE2 may act in conjunction to disrupt the BBB during experimental meningitis. PMID- 9744293 TI - Selective serotonin reuptake inhibitors reduce the spontaneous activity of dopaminergic neurons in the ventral tegmental area. AB - Electrophysiological techniques were used to study the effects of paroxetine, sertraline, and fluvoxamine on the basal activity of dopaminergic neurons in the ventral tegmental area (VTA) of rats. Acute i.v. administrations of paroxetine (20-1280 microg/kg), sertraline (20-1280 microg/kg), and fluvoxamine (20-1280 microg/ kg) caused a slight but significant reduction in the firing rate of the VTA dopaminergic cells studied. Paroxetine produced a maximal inhibitory effect of 10 +/- 11% at the cumulative dose of 160 microg/kg. Sertraline induced a dose related inhibition of VTA dopaminergic neurons, which reached its maximum (10 +/- 7%) at the cumulative dose of 1280 microg/kg. The effect of fluvoxamine on the basal firing rate of VTA dopaminergic neurons was more pronounced as compared to that of paroxetine and sertraline, in that it produced a maximal inhibition of 17 +/- 12% at the cumulative dose of 1280 microg/kg. Acute i.v. injections of paroxetine (20-1280 microg/kg), sertraline (20-1280 microg/kg), and fluvoxamine (20-5120 microg/kg) caused a dose-dependent decrease in the basal firing rate of serotonergic neurons in the dorsal raphe nucleus (DRN). Paroxetine and sertraline stopped the spontaneous firing of serotonergic neurons at the cumulative dose of 1280 microg/kg, whereas fluvoxamine reached the same effect only at the cumulative dose of 5120 microg/kg. Pretreatment with the 5-HTA1A receptor antagonist tertatolol (1 mg/kg, i.v.) reduced the inhibitory effects of paroxetine, fluvoxamine, and sertraline on the basal activity of serotonergic neurons in the DRN. Administration of tertatolol induced a 15-fold increase in the ED50 for fluvoxamine. The antagonistic effect of tertatolol was much less evident in blocking the inhibitory action exerted by paroxetine and sertraline on the activity of serotonergic neurons. Pretreatment with tertatolol (1 mg/kg, i.v.) potentiated the inhibitory effect of fluvoxamine on the basal activity of VTA dopaminergic neurons. Tertatolol did not affect the inhibitory action exerted by paroxetine and sertraline on these neurons. It is concluded that inhibition of the basal firing rate of dopaminergic neurons in the VTA is a common characteristic of selective serotonin reuptake inhibitors (SSRIs). The effects of SSRIs on VTA dopaminergic cell activity might be relevant for their therapeutic action and may explain the origin of the reported cases of akathisia. PMID- 9744294 TI - An electron microscopic observation of the vesicular acetylcholine transporter immunoreactive fibers in the rat dorsal raphe nucleus. AB - By using immunocytochemistry with an antibody directed against the vesicular acetylcholine transporter, many cholinergic neuronal processes were found to be immunopositive in the dorsal raphe nucleus. At the electron microscopic level, most of these processes were found to be axons. The immunopositive axon terminals made synapses on immunonegative dendrites and their spines whereas rare synapses were found between the immunopositive axon terminals and the immunonegative neuronal perikarya. Occasionally, the dendrites postsynaptic to an immunopositive axon terminal also received a synapse from an immunonegative axon terminal. The synapses made by the immunopositive axon terminals were usually symmetric and had a short active zone. Fewer immunostained dendrites were found, and they usually received asymmetric synapses from nonimmunostained axon terminals. The existence of cholinergic axon terminals and the synapses made by these terminals support the physiological data indicating that acetylcholine plays a role in the pain inhibition system in the dorsal raphe nucleus. PMID- 9744295 TI - Control of cytoskeletal architecture by the src-suppressed C kinase substrate, SSeCKS. AB - Activation of protein kinase C (PKC) in many cell types results in cytoskeletal reorganization associated with cell proliferation. We previously described a new cell cycle-regulated myristylated PKC substrate, SSeCKS (pronounced essex), that interacts with the actin cytoskeleton [Lin et al., 1995, 1996]. SSeCKS shares significant homology with Gravin, which encodes kinase scaffolding functions for PKC and PKA [Nauert et al., 1997]. This article describes the cellular effects of ectopically expressing SSeCKS in untransformed NIH3T3 fibroblasts. Because the constitutive overexpression of SSeCKS is toxic [Lin et al., 1995], we developed cell lines with tetracycline (tet)-regulated SSeCKS expression. The induction of SSeCKS (removal of tet) caused significant cell flattening and the elaboration of an SSeCKS-associated cortical cytoskeletal matrix resistant to Triton X-100 extraction. Flattened cells were growth-arrested and marked by the formation of cellular projections and the temporary loss of actin stress fibers and vinculin associated adhesion plaques. SSeCKS overexpression did not affect steady-state levels of actin, vinculin, or focal adhesion kinase (FAK) but did increase integrin-independent FAK tyrosine phosphorylation. Stress fiber loss was coincident with induced SSeCKS expression, strongly suggesting a direct effect. Cytochalasin, and to a lesser extent nocodazole, inhibited SSeCKS-induced cell flattening, however, only cytochalasin affected the shape of pre-flattened cells, suggesting a greater dependence on microfilaments, rather than microtubules. By contrast, only nocodazole caused retraction of the filopodia-like processes. These data indicate a role for SSeCKS in modulating both cytoskeletal and signaling pathways. Thus, we propose to expand SSeCKS scaffolding functions to include the ability to control actin-based cytoskeletal architecture, as well as mitogenic signal pathways. PMID- 9744296 TI - The Golgi apparatus and the centrosome are localized to the sites of newly emerging axons in cerebellar granule neurons in vitro. AB - Cultured cerebellar granule neurons develop their characteristic axonal and dendritic morphologies in a series of discrete temporal steps highly similar to those observed in situ, initially extending a single process, followed by the extension of a second process from the opposite pole of the cell, both of which develop into axons to generate a bipolar morphology. A mature morphology is attained following the outgrowth of multiple, short dendrites [Powell et al., 1997: J. Neurobiol. 32:223-236]. To determine the relationship between the localization of the Golgi apparatus, the site of microtubule nucleation (the centrosome), and the sites of initial and secondary axonal extension, the intracellular positioning of the Golgi and centrosome was observed during the differentiation of postnatal mouse granule neurons in vitro. The Golgi was labeled using the fluorescent lipid analogue, C5-DMB-Ceramide, or by indirect immunofluorescence using antibodies against the Golgi resident protein, alpha mannosidase II. At 1-2 days in vitro (DIV), the Golgi was positioned at the base of the initial process in 99% of unipolar cells observed. By 3 DIV, many cells began the transition to a bipolar morphology by extending a short neurite from the pole of the cell opposite to the initial process. The Golgi was observed at this site of secondary outgrowth in 92% of these "transitional" cells, suggesting that the Golgi was repositioned from the base of the initial process to the site of secondary neurite outgrowth. As the second process elongated and the cells proceeded to the bipolar stage of development, or at later stages when distinct axonal and somatodendritic domains had been established, the Golgi was not consistently positioned at the base of either axons or dendrites, and was most often found at sites on the plasma membrane from which no processes originated. To determine the location of the centrosome in relation to the Golgi during development, granule neurons were labeled with antibodies against gamma-tubulin and optically sectioned using confocal microscopy. The centrosome was consistently co-localized with the Golgi during all stages of differentiation, and also appeared to be repositioned to the base of the newly emerging axon during the transition from a unipolar to a bipolar morphology. These findings indicate that during the early stages of granule cell axonal outgrowth, the Golgi centrosome is positioned at the base of the initial axon and is then repositioned to the base of the newly emerging secondary axon. Such an intracellular reorientation of these organelles may be important in maintaining the characteristic developmental pattern of granule neurons by establishing the polarized microtubule network and the directed flow of membranous vesicles required for initial axonal elaboration. PMID- 9744297 TI - Spermiogenesis in Marsilea vestita: a temporal correlation between centrin expression and blepharoplast differentiation. AB - The motile male gamete of the water fern Marsilea vestita is a spirally shaped cell that possesses a complex cytoskeletal array of microtubules and approximately 140 cilia. Spermiogenesis in this organism is a rapid process that requires only approximately 11 h at 20 degrees C and involves the de novo synthesis of basal bodies from an organelle known as a blepharoplast. The developmental program that gives rise to the spermatozoids begins with nine mitotic divisions that occur in rapid succession during the first 5.5 h after imbibition of the dry microspore. During the next 5.5 h, the spermatids undergo a complicated differentiation process. We have asked what new proteins must be made for differentiation to proceed. Inhibitor treatments reveal that some translation is a necessary prerequisite for the differentiation and release of spermatozoids, but methionine-labeling studies demonstrate that relatively few types of proteins must be translated for this developmental program to reach completion. We have found that the dry microspores contain alpha-, beta-, and gamma-tubulin, at levels that may be sufficient for the entire developmental process. The abundance of the tubulins remains essentially constant until very late stages of spermiogenesis. In contrast to the tubulins, we show that centrin begins to increase in abundance at approximately 4 h after imbibition and that it reaches a peak at 6 h after imbibition. We also show that centrin mRNA is stored in the dry microspore, and that centrin protein abundance is regulated at the translational level. We believe that the translation of stored centrin transcripts serves as a rate-limiting step in the rapid differentiation process of spermiogenesis M. vestita. We suggest that centrin functions in the microtubule organizing centers that are required for the construction of the cytoskeleton and the motile apparatus in these structurally complicated cells. PMID- 9744298 TI - Real-time observation of Ca2+-induced basal body reorientation in Chlamydomonas. AB - The two basal bodies of Chlamydomonas are connected by a bridge, the distal fiber, that contains a Ca2+-binding protein, centrin. Although various fibrous structures in many organisms containing centrin or similar proteins have been shown to contract at Ca2+ concentrations >10(-7)-10(-6) M, the contractility of the distal fiber in Chlamydomonas has not been demonstrated. To determine whether it undergoes Ca2+-dependent contraction, we isolated the flagella-basal body complex from the paralyzed-flagella mutant pf18 and measured the angle between the two axonemes at different Ca2+ concentrations. Use of a double mutant with the mutant fa1, deficient in the mechanism for Ca2+-dependent flagellar amputation, enabled the measurement at Ca2+ concentrations > or = 10(-4) M. The angle, 80-120 degrees at 10(-9) M Ca(2-), was found to decrease by about 20 degrees when the Ca2+ concentration was raised above 10(-6) M. The angle increased again when the Ca2+ concentration was lowered below 10(-7) M. The flagellar apparatuses isolated from the double mutant between pf18 and the mutant vfl2 deficient in the structural gene of centrin had an angle of 90-130 degrees at 10(-9) M Ca2+, but the angle did not change when the Ca2+ concentration was increased. Thus centrin must be involved in the basal body reorientation. In detergent-extracted cell models of the pf18fa1 mutant, the angle between the two axonemes was found to decrease transiently by about 15 degrees upon iontophoretic application of Ca2+. Hence, the Ca2+-induced basal body reorientation can take place even when the basal body is contained in the cell body covered by the cell wall. It may function as part of the mechanism for phobic responses wherein Chlamydomonas cells swim backward transiently upon reception of strong light or mechanical stimuli. PMID- 9744299 TI - Overexpression of the 77-kD echinoderm microtubule-associated protein (EMAP), a WD-40 repeat protein, in baculovirus-infected Sf9 cells. AB - The purpose of this study was to test whether any assembly-promoting microtubule associated protein (MAP) would bundle microtubules and induce process formation in recombinant baculovirus-infected Sf9 cells, in particular, whether a non neural MAP from a normally rounded cell would produce cellular asymmetries. To carry out these experiments, we constructed a recombinant baculovirus that expressed the full-length 77-kD EMAP, an abundant MAP that localizes to the mitotic spindle of cleavage-stage sea urchin embryos and to the interphase array of microtubules in adult coelomocytes. Expression of EMAP in Sf9 cells had no detectable effect on cellular morphology, microtubule organization, or stability. These results indicate that process formation in Sf9 cells is MAP specific. PMID- 9744300 TI - Alteration of fibronectin affinity during differentiation modulates the in vitro migration velocities of Hydra nematocytes. AB - In the fresh water Cnidarian Hydra nematocytes differentiate from stem cells in the body column of the polyps and are functional in the tentacles to where they migrate as single cells in an amoeboid fashion. The fluorescent vital stain TROMI (tetramethyl-rhodamine-5/6-maleimide) allows to easily discriminate between cells located in the body column and cells mounted in the tentacles. The two cell populations were found to have different in vitro migration properties. These differences appear to be due largely to a differential attachment to fibronectin. Nematocytes from the tentacles show significantly lower in vitro migration velocities on isolated pieces of the organisms extracellular matrix (the mesoglea) and attach more firmly to fibronectin-coated substrates than cells from the body column. Pretreatment of the mesogleae with antibodies against the cell binding domain of fibronectin or addition of RGD-peptides results in an increase of the average migration velocity of cells from the tentacles and a decreased velocity of the cells from the body column. These findings suggest that (1) modulation of the attachment to fibronectin is decisive for the observed differential migration properties of the two cell populations and (2) the in vitro migration of nematocytes is dependent on subtle and transient interactions of cell surface receptors (most probably integrins) and fibronectin. PMID- 9744302 TI - Adhesion molecules and tumor metastasis. PMID- 9744301 TI - Association of spectrin with a subcompartment of the endoplasmic reticulum in honeybee photoreceptor cells. AB - The endoplasmic reticulum (ER) in honeybee photoreceptors is organized into structurally distinct subregions. The most prominent of these, the submicrovillar network of ER cisternae, is tightly associated with actin filaments. Electron microscopic techniques have demonstrated that the ER-associated actin filaments are regularly spaced at 60-80 nm and cross-bridged by filamentous structures. A polyclonal antibody against Drosophila alpha-spectrin has been used to examine the distribution of spectrin in the photoreceptors. On Western blots of bee retina, the antibody identifies a 260-kDa protein that exhibits biochemical and immunological properties characteristic of alpha-spectrin. Immunofluorescence microscopy has shown that alpha-spectrin codistributes with the submicrovillar ER but not with other ER subdomains. After cytochalasin-B-induced depolymerization of the ER-associated F-actin system, alpha-spectrin remains colocalized with the ER, indicating that alpha-spectrin is bound to the ER membrane. The F actin/spectrin system associated with the submicrovillar ER may stabilize the shape of this ER subcompartment and may play a role in maintaining functional ER subregions. PMID- 9744303 TI - Recommendations for the reporting of pancreatic specimens containing malignant tumors. Association of Directors of Anatomic and Surgical Pathology. PMID- 9744304 TI - VCAM (IGSF) adhesion molecule expression in breast carcinomas detected by automated and quantitative immunocytochemical assays. AB - Expression of vascular cell adhesion molecules (VCAM) in tumors is associated with endothelial cell activation and may facilitate adherence of carcinomatous cells to the vessel wall, promoting bloodborne metastases. Expression of VCAM was investigated in 202 breast carcinomas using automated (Ventana System) and quantitative (SAMBA image analyzer) immunoperoxidase staining of frozen sections. Positive VCAM immunoreactivity was observed in 83 tumors (41%) (mean immunostained surface, 12.4%; SD, 10.5). The mean area of immunostaining was correlated with clinical and pathologic prognostic indicators and with the immunohistochemical expression in tissue sections of various indicators of cell proliferation, metastatic potential, and drug resistance or sensitivity, evaluated according to the same method. There was no correlation of VCAM immunoreactivity with tumor size, type, or grade or with nodal status. Also, no significant correlation was observed between VCAM and MIB1/Ki67, p53, Bcl-2, E cadherin, CD44v, cathepsin D, CD31, P-gp, ER, PR, or pS2. However, VCAM immunoreactivity was significantly correlated with ELAM and VLA2 (P = .001) and VLAs (P = .008) expression. The results suggest that VCAM expression in breast carcinoma tissue sections is likely not a prognostic indicator. Its practical clinical relevance, if any, must be established by correlation with patients' outcomes and tumor sensitivity to drugs. PMID- 9744305 TI - Hamartoma of mature cardiac myocytes. AB - The clinical and pathological findings of three patients with hamartomas of mature cardiac myocytes resembling localized hypertrophic cardiomyopathy are presented. Hypertrophic cardiomyopathy is manifest by a poorly demarcated area of cardiac hypertrophy, microscopically demonstrating myofiber disarray and intramural coronary thickening. Localized, nonencapsulated masses of hypertrophied cardiac myocytes in locations other than the left ventricle or ventricular septum have not been reported. The clinical and pathological data of three patients with localized hamartomas were retrospectively retrieved. The patients were 9, 22, and 28 years old, respectively; none had a known family history of heart disease or cardiomyopathy. Two patients had cardiac arrhythmias: one patient died suddenly, and one patient had the Wolff-Parkinson-White syndrome. The third patient was asymptomatic. Two patients treated surgically had single masses in the right atrium and right ventricle, respectively. The patient who died suddenly had multiple discrete masses throughout the atrial and ventricular myocardium, including the left ventricular free wall. None of the three patients had septal asymmetry suggestive of hypertrophic cardiomyopathy. Histologically, there were discrete but unencapsulated nodules of marked myocyte hypertrophy with disorganization, focal scarring, and thickened intramural arteries. There was no myocyte vacuolization suggestive of cardiac rhabdomyoma. Ultrastructurally, the myocytes showed abundant and disorganized myofilaments and normal intercellular junctions. Hamartoma of mature cardiac myocytes is a previously undescribed cardiac tumor that shares some features of hypertrophic cardiomyopathy and rhabdomyoma, but is currently best considered a separate entity. PMID- 9744306 TI - Tissue-specific expression pattern of vascular endothelial growth factor isoforms in the malignant transformation of lung and colon. AB - Angiogenesis, a prerequisite for tumor growth and progression, results from a shift in the equilibrium between angiogenic factors and angiogenic inhibitors. Vascular endothelial growth factor (VEGF) has been identified as one of the most important factors mediating angiogenesis in physiological and pathological conditions. Through alternative splicing, four isoforms of VEGF are formed, consisting of 206, 189, 165, and 121 amino acids, respectively. VEGF206 and VEGF189 differ from VEGF165 and VEGF121 in their bioavailability, with the longer forms being matrix-bound and the shorter forms freely diffusible. To investigate the relative importance of the VEGF isoforms in neoplastic transformation, we studied the pattern of splice variant expression by reverse transcription polymerase chain reaction (RT-PCR) in 18 lung and 11 colonic carcinomas and their corresponding normal tissues, respectively. The findings showed a significant upregulation of VEGF in both carcinomas, with VEGF165 and VEGF121 being the predominant forms; VEGF189 was significantly expressed in normal lung but not colon; and VEGF206 was not detected in any specimen. The findings indicate that during malignant progression, an angiogenic switch favoring the shorter diffusible isoforms is likely to confer on the tumor a growth advantage. From the differential expression of VEGF isoforms in normal lung and colonic tissues, different functional roles of the splice variants is suggested. In particular, VEGF189 may be important for the maintenance of the vascular integrity of the lung. PMID- 9744307 TI - Prediction of local recurrence of ductal carcinoma in situ of the breast using five histological classifications: a comparative study with long follow-up. AB - The increased detection of ductal carcinoma in situ (DCIS) by mammographic screening and the more widespread use of breast-conserving surgery have led to a search for histological features associated with the risk of recurrence. In a case control study of 141 patients with long follow-up, we compared the ability of five morphological classifications to predict recurrence after local excision. A significant correlation was not found between recurrence and growth pattern when a traditional classification based on architecture was used nor with necrosis when a scheme based principally on this feature was employed. A correlation was, however, found between recurrence and "differentiation" as defined by nuclear features and cell polarization in a classification recently formulated by the European Pathologists Working Group (EPWG), but this failed to reach statistical significance at the 5% level. A stronger and statistically significant correlation was found between nuclear grade as defined by the EPWG and recurrence when cell polarization was disregarded, using the classification currently employed by the UK National Health Service and European Commission funded Breast Screening Programmes. This was attributable to a small number of recurring cases being downgraded as a consequence of exhibiting polarized cells. A significant correlation between histology and recurrence was also observed using the Van Nuys classification, which is based on nuclear grade and necrosis. Whether the tumor recurred as in situ or invasive carcinoma was unrelated to histological classification, as was the time course over which it occurred. These findings strongly support the use of nuclear grade to identify cases of DCIS at high risk of recurrence after local excision, but further work is necessary to determine whether nuclear grade or necrosis is more appropriate to subdivide the non-high-grade cases. PMID- 9744308 TI - Comparison of estrogen and progesterone receptor, Ki-67, and p53 immunoreactivity in uterine endometrioid carcinoma and endometrioid carcinoma with squamous, mucinous, secretory, and ciliated cell differentiation. AB - An analysis of 77 uterine endometrioid carcinomas was performed to compare pure endometrioid carcinomas and endometrioid carcinomas with various types of cellular differentiation for the expression of estrogen (ER) and progesterone (PR) receptors, p53, and Ki-67 and to correlate these findings with clinicopathologic features. Forty-three pure endometrioid carcinomas and 34 endometrioid carcinomas displaying additional types of cellular differentiation in at least 10% of the tumor (16 squamous, 11 mucinous, four ciliated cell, and three secretory) were analyzed. In 8 of the 16 tumors with squamous differentiation, the squamous component was histologically benign (low grade), and in eight tumors it was histologically malignant (high grade). In tumors showing various types of cellular differentiation except those with a high-grade squamous component, comparison of the endometrioid glandular component with the squamous, mucinous, secretory, and ciliated cell components showed that ER/PR, Ki 67, and p53 expression were generally higher in the glandular component compared with the various differentiated components. These findings parallel the changes that occur in the endometrium in the secretory phase of the menstrual cycle and, therefore, suggest that the differentiated components have undergone terminal differentiation. In contrast, in endometrioid carcinomas with a high-grade squamous component, Ki-67 and p53 expression were the same in the glandular and squamous components suggesting that squamous epithelium in these tumors represented another pathway of cellular differentiation but not one that was terminally differentiated. Endometrioid carcinomas with a high-grade squamous component had significantly higher grade (P = .002), stage (P < .001), cellular proliferation index (P = .0005), and worse outcome (P = .0009) compared with tumors with the other types of cellular differentiation, including those with a low-grade squamous component and pure low-grade endometrioid carcinomas. In addition, carcinomas with a high-grade squamous component occurred in older women and were more frequently associated with atrophic endometrium and less replacement hormone therapy, but the differences were not statistically significant. In conclusion, endometrioid carcinomas with various types of cellular differentiation can be broadly divided into two groups. Tumors with mucinous, secretory, and ciliated cell differentiation and those with a low-grade squamous component are similar to pure low-grade endometrioid carcinomas in that most have high ER and PR expression, low cellular proliferation indices, low p53 immunoreactivity, and good prognosis. In contrast, endometrioid carcinomas with a high-grade squamous component lack expression of ER and PR, have high cellular proliferation indices, often express p53, and have a prognosis similar to poorly differentiated endometrioid carcinomas. PMID- 9744309 TI - Vulvar lichen sclerosus and squamous cell carcinoma: a cohort, case control, and investigational study with historical perspective; implications for chronic inflammation and sclerosis in the development of neoplasia. AB - The histological changes of lichen sclerosus (LS) are frequently found in association with vulvar squamous cell carcinoma (SCC). The importance of chronic inflammation and scarring in oncogenesis is well recognized. Thirty-two patients with symptomatic vulvar LS and 60 with vulvar SCC were studied. Paraffin sections of vulvar LS, and three controls groups (acute scars, normal vulva, and vulvar lichen simplex chronicus [LSC]) were investigated with a panel of seven tissue markers and for DNA content in areas without vulvar intraepithelial neoplasia (VIN). All published cases to date of vulvar LS associated with SCC were reviewed. Of the cohort of symptomatic vulvar LS patients (mean/median age, 60 years), 9% developed VIN lesions and 21% invasive SCC; symptomatic LS preceded the carcinoma by a mean of 4 years (range, 1 to 23 years). Second and third primary tumors developed in three of these patients. Of the series of 60 patients presenting with vulvar SCCa, the clinical setting and histological features of SCCs associated with LS were significantly distinctive compared with SCCas without LS: SCCs associated with LS occurred in an older age-group (74 v 65 years; P = .01), were located on the clitoris (41% v 5%; P = .003), were of conventional SCCa type (85% v 57%; P = .02), were associated with a prominent fibromyxoid stromal response (46% v 10%; P = .004), were not associated with VIN 3 (SCC in situ) (5% v 67%; P = .02) and diffusely expressed tumor suppressor gene product p53 (43% v 19%; P = .01) and cytokine TGF-beta (33% v 9%; P = .05). The epidermis of vulvar LS was similar to that of acute scars and differed significantly compared with normal vulva with respect to keratinocytic expression of markers to keratin AE 1, involucrin and filaggrin, epidermal thickness (0.13 mm [LS] v 0.05 mm [normal]; P < .03), and proliferative index by PCNA and Mib-1 labeling (53/60 [LS] v 15/19 [normal] per 200 basal cells [bc]; P < .003). Vulvar LS showed significantly higher expression of p53 than all three control groups (80 [LS] v 3 [normal]/44 [acute scar]/28 [LSC] per 200 bc; P < .008), and aneuploidy (33% v diploid controls) in the absence of VIN. Comparing LS with and without associated SCCa found significant increases in age of patients (74 v 66 years; P = .001), and DNA aneuploidy (52% v 11%; P = .0001) and no differences in epidermal thickness, sclerotic thickness, proliferative index, or p53 expression. However, those cases of LS with an aneuploid DNA content showed significantly elevated p53 expression (88 v 60/200 bc; P = .01) and epidermal thickness (0.16 v 0.11 mm; P = .005) compared with LS with a diploid DNA content. Review of published cases supports an association between LS and vulvar SCC. The phenomenon of chronic inflammation and scarring giving rise to carcinoma has been well documented. Vulvar lichen sclerosus (LS) is an inflammatory dermatosis characterized by clinicopathologic persistence and hypocellular fibrosis (sclerosis). A subset of vulvar SCCs is significantly associated with the presence of LS and diffusely express the p53 gene product. Keratinocytes affected by LS show a proliferative phenotype and can exhibit markers of neoplastic progression such as increased p53 expression and DNA aneuploidy. As a chronic scarring inflammatory dermatosis, vulvar LS could act as both "initiator and promoter" of carcinogenesis, explaining the frequent coexistence of these diseases. Because keratinocytes of LS significantly express tumor suppressor gene p53 protein, the p53 gene may be involved early in this proposed pathway of carcinogenesis. PMID- 9744310 TI - Ki67 labeling index in core needle biopsies independently predicts tumor-specific survival in prostate cancer. AB - A better knowledge of the biological aggressiveness of individual tumors could facilitate the selection of treatment in prostate cancer patients. This study assesses the influence of histological and molecular features in core needle biopsy specimens of prostate cancer on tumor-specific survival. Formalin-fixed core needle biopsy specimens from 111 consecutive patients (mean follow-up, 5.0 years) were immunohistochemically examined for their proliferative activity (Ki67 labeling index [LI]) and expression of p53 and Bcl-2. Overexpression of p53 was found in 16% of the biopsy specimens and was mainly restricted to poorly differentiated tumors. Bcl-2 positivity was found in 20% of tumors. The median Ki67 LI was 7.5%. There was a strong relationship between Ki67 LI and Gleason grade, with a continuous increase in the proliferative activity from low-grade to high-grade tumors (P = .0006). Univariate tumor-specific survival analysis showed that high Gleason score (P = .0018), high percentage of biopsy tumor involvement (P = .0227), high Ki67 LI (P = .0007), and p53 positivity (P = .0024) were predictors of tumor-related death. A high Ki67 LI emerged as the only independent predictor of tumor-specific survival in multiparametric analysis. These results indicate that core needle biopsy specimens of the prostate not only are useful for diagnosis of malignancy but also can provide valuable prognostic information. Immunohistochemical examinations of molecular features may be a helpful adjunct for a better pretherapeutic assessment of prostate cancer aggressiveness and therefore contribute to an improved initial patient management. PMID- 9744311 TI - p53 mutagenesis in Klatskin tumors. AB - Mutagenesis of the p53 tumor-suppressor gene represents the most common genetic alteration in human malignancies but has not yet been investigated in Klatskin tumors. Cancerous and normal liver tissues were obtained from 12 patients after surgical resection of Klatsin tumors. Genomic DNA was extracted and served as a template for PCR amplification and sequencing of a 1,574-bp fragment of the p53 gene comprising the exons 5 through 8. Immunohistochemical expression analysis was performed using five different antibodies. Missense mutations were detected in 2 of 12 patients--one transversion on codon 273 (Arg --> Leu) and a transition on codon 168 (His --> Arg). In all specimens, immunohistochemistry was negative regarding a nuclear overexpression. An apparent clinicopathologic impact of p53 mutations was not observed. This report on mutagenesis of the p53 gene in Klatskin tumors shows that the most commonly mutated tumor suppressor gene in human cancers is also mutated in a subset of patients with Klatskin tumors. Assessment of a clinical or pathological impact of p53 mutagenesis on Klatskin tumors requires evaluation in larger studies. PMID- 9744312 TI - Loss of heterozygosity of the von Hippel Lindau gene locus in polypoid dysplasia but not flat dysplasia in ulcerative colitis or sporadic adenomas. AB - Carcinoma in ulcerative colitis (UC) develops from dysplastic precursor lesions, which include flat dysplasia (FD) and polypoid dysplasias (PD). PD may present as single or multiple polypoid structures or as plaque-like lesions that, independent of histological grade, are an indication for colectomy. PDs are histologically similar to adenomas and may not be readily distinguished by light microscopy. It is not known whether FD and PD are different entities, or whether they represent etiologically similar lesions with different morphological expression. We microdissected 25 cases of UC with PD and 19 samples of FD with surrounding chronic colitis (CC) in UC. Loss of heterozygosity (LOH) at the von Hippel Lindau (vHL) gene locus and the putative tumor suppressor genes APC, INK4A (9p16), and p53 was studied. LOH of the vHL gene, INK4A (9p16), and APC was also studied in 11 sporadic adenomas of the colon. LOH at the vHL locus was present in 50% of the samples of PD and in 12% of the samples of FD. LOH was seen in CC close to PD and FD in 26% and 12% of cases, respectively. No adenoma showed LOH of the vHL gene markers studied. LOH in p53 was seen in PD in 16% cases and in FD in 42% cases and in CC close to PD and FD in 0% and 14% cases, respectively. LOH patterns between PD and FD of the markers for APC and 9p16 were not different. LOH in APC was seen in two of five cases of adenoma. We conclude that PD and FD share genetic alterations in APC and 9p16 genes. More frequent involvement of the VHL gene in PD and surrounding CC and involvement of p53 in HGD and CC in FD may represent genetic differences between the development of PD and FD and may be the cause of the different morphology. The infrequency of LOH at the vHL locus in adenomas versus PD may serve as a discriminator between adenomas and PD in diagnostically problematic cases. PMID- 9744313 TI - Differentiation and programmed cell death-related intermediate biomarkers for the development of non-small cell lung cancer: a pilot study. AB - Fifty samples of lung tissue from patients with non-small cell lung cancer were analyzed for the expression and localization of biomarkers related to squamous differentiation and programmed cell death. These markers include tissue transglutaminase (tTG), keratinocyte transglutaminase (kTG), involucrin, loricrin, and Bcl-2. We found that all of these markers are overexpressed in tumors as compared with histologically normal lung epithelium, where expression is minimal. Expression of the oncoprotein, Bcl-2, increased starting in squamous metaplasia and remained elevated in all lesions, including frank carcinoma. In contrast, expression of the other markers was elevated in the histologically abnormal noninvasive lesions but was decreased somewhat in invasive malignancy. In addition, we found that tTG, kTG, and Bcl-2, when expressed, were detected in mutually exclusive areas. These findings suggest that (1) these markers may prove useful, with more extensive testing and clinical correlation, in predicting risk for the development of lung cancer; and (2) pulmonary carcinogenesis may result from the failure of differentiation and programmed cell death mechanisms in the presence of oncogene overexpression rather than through oncogene/tumor suppressor gene abnormalities alone. PMID- 9744314 TI - Colorectal inflammation and increased cell proliferation associated with oral sodium phosphate bowel preparation solution. AB - Evidence is emerging that sodium phosphate (NaP), a commonly used oral cathartic agent, causes aphthoid ulcers or focal active colitis (FAC) in the colon and rectum. The aims of this study were (1) to assess the incidence of such ulcers diagnosed endoscopically ("aphthoid ulcers"), (2) to assess the incidence of histologically detected FAC and neutrophilic infiltration overlying lymphoid follicles ("aphthoid lesions"), and (3) to determine whether this effect of NaP is associated with epithelial cell proliferation. Aphthoid ulcers, unexplained by other diagnoses, were found in 18 of 687 consecutive patients (2.6%) who underwent colonoscopic examination after oral NaP preparation during a 12-month period; biopsy specimens showed FAC or aphthoid lesions. FAC was present in 11 of 316 patients (3.5%) who had biopsies but were endoscopically normal. Eight patients with aphthoid ulcers in the rectosigmoid showed no abnormalities when reexamined by flexible sigmoidoscopy after an interval as short as 7 days (range, 7 to 56 days). Mucosal biopsy specimens from these patients were assessed for apoptosis and epithelial proliferation by determining the MIB-1 labeling index (LI). The LI was increased by 136% after NaP preparation (55 +/- 6) compared with biopsy specimens obtained from the same patients during reexamination without NaP preparation (23 +/- 6, P = .01). This correlated with the number of apoptotic bodies per 10 colonic crypts (1.2 +/- 0.3 v 0.5 +/- 0.2, respectively). To determine whether these proliferative changes represent a response to mucosal ulceration, rectosigmoid biopsy specimens were compared in two additional patient groups: an NaP group in whom no gross lesions were evident and a no-NaP group who were not exposed to NaP. Although more modest, similar changes in the LI (42 +/- 4 and 30 +/- 3, respectively, P = .03) and in the occurrence of apoptotic bodies per 10 colonic crypts (1.3 +/- 0.4 and 0.4 +/- 0.1, respectively) were observed. We conclude that use of NaP is associated with increased colorectal crypt epithelial cell proliferation. This proliferative response to NaP exposure is evident in the absence of colonoscopically or other histologically recognizable abnormalities. In a proportion of patients, aphthoid ulcers, FAC, or aphthoid lesions serve as markers of mucosal damage by NaP. PMID- 9744315 TI - Prognostic significance of stromelysin 3, gelatinase A, and urokinase expression in breast cancer. AB - This study was aimed at testing the hypothesis that the expression of proteases essentially produced by reactive stromal cells (stromelysin-3 [ST3], gelatinase A [GELA], and urokinase [uPA]) is predictive of prognosis in patients with breast cancer. This was a study of patients with node-positive and node-negative breast cancer diagnosed from 1980 to 1986 and with an average of 10 years follow-up. ST3 (665 cases), GELA, and uPA (575 cases each) expression was obtained by in situ hybridization on formalin-fixed, paraffin-embedded material using mRNA antisense probes. ST3 was expressed by 86.6% of the cases; GELA, 77.7%; and uPA, 64.7%. A significant correlation (P < .05) was found between high (more than 10%) ST3 expression and a younger age, lymph node involvement, poor nuclear grade, ductal histology, aneuploidy, and HSP-27 expression. High GELA expression was significantly associated with c-erbB2, ductal histology, and HSP-27 expression. High uPA expression correlated with poor nuclear grade, ductal histology, lack of estrogen and progesterone receptors, and p53 protein accumulation. High level of expression of all three proteases correlated significantly with each other and with cathepsin D expression by reactive stromal cells. By univariate analysis, both ST3 and uPA expression significantly predicted a shorter recurrence-free survival (ST3, P = .0199; uPA, P = .0269). By multivariate analyses, the prognostic significance was lost, most particularly at longer term. This study adds support to the concept that protease expression by reactive stromal cells is related to cancer cell characteristics but that their contribution to cancer progression is marginal. PMID- 9744316 TI - Increased expression of vascular endothelial growth factor is associated with tumor progression in hepatocellular carcinoma. AB - Vascular endothelial growth factor is a potent direct-acting angiogenic factor. Early in hepatocarcinogenesis, hepatocellular carcinomas do not show hypervascularity; at later stages, they require abundant arterial blood flow. We investigated the role of vascular endothelial growth factor in hepatocellular carcinoma arterialization. We studied 51 patients with hepatocellular carcinoma. All patients had undergone hepatic arteriography. Vascular endothelial growth factor expression was investigated by immunohistochemistry (n = 51) and in situ hybridization (n = 13), and the changes in vascular endothelial growth factor expression were evaluated in relation to tumor differentiation and changes in tumor vascularity. The expression of vascular endothelial growth factor isoforms in hepatocellular carcinomas was also analyzed by reverse transcriptase polymerase chain reaction (n = 10). Vascular endothelial growth factor expression was detected in hepatoma cells and hepatic stellate cells, and increased vascular endothelial growth factor expression was associated with tumor dedifferentiation. Vascular endothelial growth factor expression in hypervascular hepatocellular carcinomas was greater than in those not showing hypervascularity. The major vascular endothelial growth factor isoforms expressed in hepatocellular carcinoma were 121 and 165. These findings indicate that vascular endothelial growth factors 121 and 165 play a critical role in the process of angiogenesis in hepatocellular carcinomas. PMID- 9744317 TI - Immunohistologic analysis of gastrointestinal and pulmonary carcinoid tumors. AB - Carcinoid tumors are potentially malignant neoplasms that arise in various body sites, including the lung and gastrointestinal tract. Those that appear cytologically atypical are more likely to behave aggressively than more typical carcinoid tumors. However, in the absence of cytological atypia or large tumor size, it is difficult to predict the biology of an individual tumor, because some lesions metastasize, whereas others do not. This study had four aims: (1) To study the expression pattern of p53, Ki-67, NCAM, and S-100 in carcinoid tumors and to relate these expression patterns to classical histopathologic features and to tumor location. (2) To identify nonhistological markers that might more accurately predict the early behavior of carcinoid tumors. (3) To determine whether sustentacular cells are present in carcinoid tumors arising in tissues derived from different embryological derivatives. (4) To determine the synaptophysin and chromogranin immunoreactivity in neuroendocrine tumors arising in various locations. The immunostaining reactions were quantitatively scored by three observers. Only 3 of the 39 tumors (all histologically atypical) were strongly positive for Ki-67; two of these were also strongly p53 immunoreactive. NCAM immunostaining differed according to the site of origin: 76.5% of foregut lesions, 58% of the midgut lesions, and 20% of hindgut lesions were positive. S 100 immunostaining ranged from 41% in foregut lesions to 50% in both the hindgut- and midgut-derived tumors. S-100-positive sustentacular cells were present in 20.5% of carcinoid tumors. All tumors stained with antibodies against synaptophysin. In contrast, 100% of midgut, 60% of hindgut, and 88% of foregut tumors were chromogranin positive. Carcinoid tumors tend to have low proliferative rates. p53 immunostaining tends to be strongly positive in tumors that are histologically atypical, but it is negative in typical carcinoid tumors arising in the gastrointestinal tract and lungs. Immunostaining reactions with antibodies to NCAM, S-100, and chromogranin differ depending on the site of origin. Synaptophysin stains 100% of carcinoid tumors regardless of their site of origin. In contrast, antibodies to chromogranin fail to stain 40% of hindgut tumors and 12% of foregut carcinoid tumors. S-100-positive sustentacular cells are present in foregut and midgut tumors but not in hindgut tumors. PMID- 9744318 TI - Histological parameters as predictors of prognosis in childhood brain tumors. AB - Using histological parameters with high recognition reliability, we retrospectively analyzed all newly diagnosed patients under the age of 16 years (n = 100) with brain and spinal cord tumors presenting to the National Neuroscience Centres of the Richmond and Beaumont Hospitals, Dublin, Ireland, between 1985 and 1990, allowing analysis of 5-year survival in all cases. Tumor histology was reviewed by two neuropathologists blinded to previous histological diagnosis and to the site of lesion. We found that certain histological features such as very low cell density and microcyst formation had a positive effect on prognosis. Mitoses and pleomorphism had a negative effect on prognosis, whereas necrosis and meningeal involvement had no effect on prognosis. It is suggested that identification of reliably recognized histological features rather than assignation of tumors to particular diagnostic categories may be a more reliable predictor of tumor behavior in the pediatric age-group. PMID- 9744319 TI - Modulation of cytokeratin subtype, EGF receptor, and androgen receptor expression during progression of prostate cancer. AB - After initial regression in response to androgen deprivation, most prostate cancers develop resistance to endocrine therapy. Identification of cellular and molecular changes occurring during endocrine therapy-induced regression and subsequent hormone insensitivity may point to mechanisms underlying the transition to hormone-independent prostate cancer. A series of untreated (n = 24), regressed (n = 15), and endocrine therapy-resistant (n = 10) prostatic adenocarcinomas were analyzed using immunohistochemistry with regard to cytokeratin 5 and 18, androgen receptor (AR), and epidermal growth factor receptor (EGF-R) expression in tumor cells. Using semiquantitative reverse transcription-polymerase chain reaction, the amount of AR mRNA also was determined. In regressed and therapy-resistant prostate cancers, an increase in cytokeratin 5-positive tumor cells was noted when compared with untreated carcinomas. Similarly, the proportion of EGF-R-positive tumor cells increased in the treated cases, whereas the proportion of AR-positive tumor cells dropped in regressed carcinomas and increased in hormone-refractory cancers. In the latter group, an eightfold higher level of AR mRNA was observed when compared with the other cases. Changes in the proportion of cytokeratin 5 and EGF-R-positive tumor cells suggests that during androgen deprivation an enlarged subpopulation of tumor cells with combined features of basal and secretory phenotypes arises. The increased proportion of AR-positive tumor cells during the transition from the regression phase to the hormone escape phase points to an important role of AR overexpression in this process. PMID- 9744320 TI - DNA ploidy by image cytometry and karyotype in spontaneous abortion. AB - We compared the DNA content (DI) by cell image analysis with the karyotype and morphological phenotype of paraffin-embedded tissues from 51 spontaneous abortions. The study included 21 cases with triploid, 19 cases with diploid, and 11 cases with aneuploid (monosomic, trisomic, or mosaic) karyotype. Measurements were performed by image analysis on the trophoblastic and stromal cells of chorionic villi using 5-microm-thick, Feulgen-stained sections. At least 200 cells were analyzed. Results were interpreted using DI ranges of 1.3 to 1.7 for triploid and 0.9 to 1.1 for a diploid profile. All 21 cases with a cytogenetically confirmed triploid karyotype had DI values within the triploid range, and all 19 cases with a diploid karyotype had DI values within the diploid range. All of the trisomies and monosomies also had a DNA mass within the diploid range. However, eight cases with a triploid karyotype also had a peak in the diploid range: one case with a diploid karyotype and one case with a trisomic karyotype each had an additional peak in the triploid range. We did not find a morphological correlation either with image analysis or with karyotype. We conclude that cell image analysis is a reliable method for detection of triploidy in spontaneous abortions. This relatively rapid method allows visual discrimination of the areas to be analyzed, avoids the problem of maternal cell contamination, and may unmask mosaic karyotypes that would go unrecognized by cytogenetic studies alone. PMID- 9744321 TI - Core morphological concepts of disease for second-year medical students. AB - With the increasing emphasis on independent learning and early patient contact, the time in the undergraduate medical curriculum for formal teaching of morphology of disease is decreasing. Thus, we thought it advisable to identify those core morphological entities of disease that should not get lost in the new paradigm. Our approach was to list all disease processes in Robbins Pathologic Basis of Diseases, 5th edition, that have distinguishing gross or microscopic characteristics. Appropriate portions of this list of 952 morphological entities from the Robbins textbook were distributed to 46 clinical specialists and pathology faculty. Each of these was asked to strongly agree, agree, disagree, or strongly disagree with the following for each entry on the list: "For purposes of developing concepts of disease, an M2 physician in training should recognize classical examples or a diagrammatic representation of the following lesions, and distinguish them from each other." Responses resulted in a consensus core list of 63 general disease process lesions and 545 organ system lesions, for a total of 608. These 608 core morphological entities were incorporated into our course by means of (1) a computer program with over 1,022 images and clinical-pathological correlations, and (2) a core list of morphological objectives for each unit in the course. In general, entities were judged noncore material if they were rare or were microscopic lesions of primary interest to pathologists and provided no major pathomorphologic concepts. The computer program as a supplement to glass slides and gross specimens has been very well accepted by students, and satisfactory performance on examinations has been maintained in spite of a 25% reduction in pathology course contact hours. PMID- 9744322 TI - B-cell lymphoma of mucosa-associated lymphoid tissue of the thymus: a report of two cases with a background of Sjogren's syndrome and monoclonal gammopathy. AB - Two rare cases of low-grade B-cell lymphoma of mucosa-associated lymphoid tissue (MALT) arising in the thymus are reported. Both patients (a 61-year-old man and a 75-year-old woman) were suffering from Sjogren's syndrome and immunoglobulin (Ig)A kappa monoclonal gammopathy. Mixed IgA-IgG cryoglobulinemia was also present in the male case. Tumor cells expressed IgA and kappa antibody reactive proteins identical with serum IgA kappa M. Moreover, we could demonstrate rearrangements of the immunoglobulin heavy and light chain genes, which supported the monoclonal origin of tumor cells. Immunological abnormalities improved after thymectomy in one case in which the tumor cells were confined to the thymus, but not the other with regional lymph node involvement, suggesting a causal role for the tumor. MALT lymphomas of the thymus thus appear to be associated with immunological disorders such as Sjogren's syndrome or monoclonal gammopathy. PMID- 9744323 TI - Cellular localization of interleukin-5 expression in rectal carcinoma with eosinophilia. AB - Eosinophilia often occurs in malignant diseases. This report concerns a female patient aged 76 years, diagnosed with rectal carcinoma with eosinophilia. Sera were obtained at two different periods (at diagnosis and after the operation) for the evaluation of levels of interleukin-5 (IL-5). The serum IL-5 level increased to 264 pg/mL, and returned to an undetectable level after the operation. The serum at diagnosis enhanced the viability of normal eosinophils in vitro, and this activity was inhibited by antihuman IL-5 polyclonal antibody. Immunohistochemistry and in situ hybridization revealed that stromal eosinophils contained IL-5 protein and messenger RNA (mRNA), but no IL-5 transcripts were detected in eosinophils attached to carcinoma cells. In situ detection of apoptosis showed that several eosinophils attached to tumor cells underwent apoptosis and lost their eosinophil secreted cationic protein (ECP) and major basic protein (MBP). These results may suggest that activated eosinophils by IL-5 play an important role in host defense mechanisms, releasing their toxic granule proteins on adjoining tumor cells. PMID- 9744324 TI - Twins, placentas, and genetics: acardiac twinning in a dichorionic, diamniotic, monozygotic twin gestation. AB - We describe a human acardiac twin with associated vascular anastomoses in a dichorionic diamniotic fused twin placenta. A 22-year-old woman delivered a healthy 3,554 g male infant and a fused diamniotic dichorionic twin placenta with a 230 g umbilical cord-attached, skin-covered, ovoid mass, consistent with acardiac amorphus. By gross and histological examination, the placental dividing membranes comprised four leaves, one amnion from each placenta, and two centrally fused chorions, diagnostic of dichorionicity. Placental barium injection of the normal twin's umbilical vein showed an anastomosis with the acardiac twin which traversed the dividing membranes, then supplied major vessels of the acardiac mass via its 5.5 cm umbilical cord. DNA-typing studies of the normal twin's placenta and of the acardiac twin's tissues revealed identical alleles at 11 distinct genetic polymorphic loci, consistent with monozygosity. Our findings demonstrate that vascular anastomoses can occur in dichorionic twin placentas, and that human acardiac twinning is not, as heretofore believed, restricted to monochorionic placentas. PMID- 9744326 TI - 1998 annual meeting press conferences produce limited coverage of nuclear medicine but reach new media outlets. PMID- 9744325 TI - Pathogenesis of pulmonary intralobar sequestration (ILS) PMID- 9744327 TI - Innovative strategies for less common thyroid cancers. PMID- 9744328 TI - Comparison of fluorine-18-FDG with rest-redistribution thallium-201 SPECT to delineate viable myocardium and predict functional recovery after revascularization. PMID- 9744329 TI - Direct evidence of impaired cardiac sympathetic innervation in essential hypertensive patients with left ventricular hypertrophy. AB - Increased sympathetic nervous activity has been proposed as one of the causes of left ventricular hypertrophy (LVH) associated with hypertension. However, the precise relationship is not fully understood. METHODS: To elucidate the relationship between myocardial sympathetic nervous activity and LVH in patients with essential hypertension EHT), we performed 123I-metaiodobenzylguanidine (MIBG) myocardial scintigraphy in 49 patients with EHT and 17 normotensive control subjects. Sympathetic innervation of the left ventricle was evaluated using SPECT, and the whole heart uptake of the tracer was quantitatively assessed as the heart-to-mediastinum uptake ratio on both the early (15-min) and delayed (5-hr) images. Myocardial washout rate (MWR) of the tracer from 15 min to 5 hr after the isotope administration was also calculated. The left ventricular mass index (LVMI) was determined echocardiographically. RESULTS: In 49 hypertensive patients, there was a negative correlation between LVMI and heart-to-mediastinum uptake ratio on both the early and delayed images (r=-0.55, p < 0.0001; r=-0.63, p < 0.0001, respectively). In addition, there was a positive correlation between the LVMI and MWR of 123I-MIBG in these hypertensive patients (r=0.59, p < 0.0001). As for the regional uptake of the tracer, there was no significant difference between control subjects and hypertensive patients without cardiac hypertrophy, but a significant decrease of the uptake in the inferior and lateral regions was observed in hypertensive patients with cardiac hypertrophy. CONCLUSION: Patients with EHT had decreased accumulation and increased MWR of 123I-MIBG in proportion to the degree of LVH. Hypertensive patients with cardiac hypertrophy had impaired sympathetic innervation in the inferior and lateral regions of the left ventricle. PMID- 9744330 TI - Outcome significance of thallium-201 and iodine-123-BMIPP perfusion-metabolism mismatch in preinfarction angina. AB - The relevance of brief antecedent ischemia to preservation of myocyte viability and cardiac function is still controversial in humans. Dysfunctioning but viable myocardium shows impaired fatty acid metabolism despite restored coronary perfusion. We asked whether preinfarction angina might be related to preservation of cell viability and better functional recovery in comparison with impaired fatty acid metabolism. METHODS: Tomographic imagings with thallium and beta methyl-p-iodophenyl penta-decanoic acid (BMIPP) were performed in 32 patients with first acute myocardial infarction who received primary coronary angioplasty: 20 patients with preexisting angina before infarction (Group A) and 12 without (Group B). Thallium and BMIPP abnormalities were quantified as a severity index by a polar map. Regional function was quantified by ventriculography and followed up. RESULTS: Despite no significant difference in coronary risk factors, cardiac function and angiographic findings, the thallium severity index was significantly lower than that of BMIPP (62+/-45 versus 96+/-59) in Group A but not in Group B (104+/-65 versus 115+/-68); the thallium severity index in Group A was significantly lower than that in Group B, but there was no significant difference in BMIPP abnormality between them. The BMIPP severity index correlated significantly with that of thallium in both groups. However, the regression line in Group A shifted downward and was statistically different compared with that in Group B. Regional function at an acute stage was significantly improved from 107+/-31 to 70+/-31 s.d./chord during follow-up in Group A but not in Group B (109+/-62 versus 106+/-52). The ratio of the thallium severity index to that of BMIPP at an acute stage was significantly related to improved regional wall motion during follow-up in the reperfused patients (y=-53x + 65, r=0.667). CONCLUSION: Preinfarction angina preserves myocyte viability relative to fatty acid metabolism, resulting in augmented perfusion-metabolism mismatch and functional improvement in patients undergoing successful reperfusion, indicating cardioprotective effects of preinfarction angina. PMID- 9744331 TI - Iodine-123-beta-CIT and iodine-123-FPCIT SPECT measurement of dopamine transporters in healthy subjects and Parkinson's patients. AB - Iodine-123-beta-carbomethoxy-3 beta-(4-iodophenyltropane) (CIT) has been used as a probe of dopamine transporters in Parkinson's disease patients using SPECT. This tracer has a protracted period of striatal uptake enabling imaging 14-24 hr postinjection for stable quantitative measures of dopamine transporters, and it binds with nanomolar affinity to the serotonin transporter. Iodine-123 fluoropropyl (FP)CIT is an analog of [123I]-beta-CIT and has been shown to achieve peak tracer uptake in the brain within hours postinjection and to provide greater selectivity for the dopamine transporter. The purpose of the present study was to compare [123I]-beta-CIT with [123I]-FPCIT in a within-subject design. METHODS: Six Parkinson's disease patients and five healthy control subjects participated in one [123I]-beta-CIT and one [123I]-FPCIT SPECT scan separated by 7-21 days. Controls were imaged at 24 hr postinjection 222 MBq (6 mCi) [123I]-beta-CIT and serially from 1-6 hr postinjection 333 MBq (9 mCi) [123I]-FPCIT. Two imaging outcome measures were evaluated: (a) the ratio of specific striatal activity to nondisplaceable uptake, also designated V"3, at each imaging time point; and (b) the rate of striatal washout of radiotracer expressed as a percent reduction per hr for [123I]-FPCIT. In addition, venous plasma was obtained from the five control subjects after the [123I]-FPCIT injection for analysis of radiometabolites. RESULTS: Both [123I]-FPCIT and [123I] beta-CIT demonstrated decreased striatal uptake in Parkinson's disease patients compared with the controls with a mean of V"3=3.5 and 6.7 for [123I]-beta-CIT (Parkinson's disease and controls, respectively) and a mean of V"3=1.34 and 3.70 for [123I]-FPCIT (Parkinson's disease and controls, respectively). For [123I] beta-CIT, the mean Parkinson's disease values represented 52% of the control uptake, while the mean [123I]-FPCIT value for Parkinson's disease patients was 37% of the control values. Analysis of [123I]-FPCIT time-activity curves for specific striatal counts showed washout rates of 8.2%/hr for Parkinson's disease and 4.9%/hr for controls. CONCLUSION: These data suggest that SPECT imaging with [123I]-FPCIT visually demonstrates reductions in striatal uptake similar to [123I]-beta-CIT. iodine-123-FPCIT washed out from striatal tissue 15-20 times faster than [123I]-beta-CIT, and estimates of dopamine transporter loss in Parkinson's disease patients were higher for [123I]-FPCIT than for [123I]-beta CIT. This was most likely due to the faster rate of striatal washout and establishment of transient equilibrium binding conditions at the dopamine transporter, which the modeling theory suggests produces an overestimation of dopamine transporter density with relatively greater overestimates in healthy control subjects by [123I]-FPCIT. PMID- 9744332 TI - Discordance of technetium-99m-HMPAO and technetium-99m-ECD SPECT in herpes simplex encephalitis. AB - Technetium-99m-hexamethyl propyleneamine oxime (HMPAO) and 99mTc-ethyl cysteinate dimer (ECD) accumulate in brain tissue in proportion to regional cerebral blood flow in healthy subjects and in patients with a variety of neurological diseases. We report on four patients with herpes simplex encephalitis and the discordance between these two approved cerebral perfusion imaging radiopharmaceuticals. CONCLUSION: SPECT images showed unilateral regional increase of 99mTc-HMPAO uptake and decrease of 99mTc-ECD uptake in the affected temporal lobe. PMID- 9744333 TI - Age-related changes in D2 receptor binding with iodine-123-iodobenzofuran SPECT. AB - The purpose of this study was to evaluate the effects of age on D2 receptor binding with 123I-iodobenzofuran (IBF) SPECT. METHODS: Subjects were 40 healthy volunteers (age 19-83 yr), including 6 who had test/retest studies. Scans were acquired with a triple-head SPECT camera 3 hr postinjection of IBF (300 MBq). Striatal regions (caudate and putamen) were defined by two different region-of interest (ROI) sets consisting of large volumes [(CLVs), 2.2 and 6.6 m] and small volumes [(SVs), 0.6 and 1.3 ml]. D2 binding (Rv=V3/V2) was quantified using our previously proposed multilinear regression technique. Effects of age on D2 binding were evaluated by fitting linear, exponential and logarithmic models. RESULTS: The mean Rvs were 26% lower than LV for both putamen and caudate than the corresponding values from the SV due to the partial-volume effect. Although the identifiability of Rv using SV deteriorated slightly, the test/retest reproducibility of Rv measurements was equally excellent for LV and SV. The mean Rvs were 11% higher for putamen compared with those for caudate. D2 binding declined significantly with age (p < 10(-5)) for all three models. The nonlinear models were slightly superior to the linear model in describing the relationship between Rv and age. In these models, D2 binding declined with age, equally for caudate and putamen at 7%-13% per decade; the decline was progressively smaller with age. CONCLUSION: IBF SPECT permitted reliable measurements of D2 binding in the caudate or putamen separately using small ROI volumes that significantly improved the quantitation loss from the partial-volume effect. Our results agreed with previous PET and postmortem findings of D2 binding losses with age. However, these age effects may be nonlinear. Age-related changes in D2 binding must be taken into consideration in clinical IBF SPECT investigations. PMID- 9744334 TI - Decreased benzodiazepine receptor binding in Machado-Joseph disease. AB - Benzodiazepine receptor binding was assessed in four Japanese men with Machado Joseph disease. METHODS: The distribution of benzodiazepine receptors was measured by radionuclide imaging (SPECT) after intravenous administration of 123I iomazenil (Ro 16-0154). RESULTS: SPECT demonstrated decreased binding throughout the cerebral cortex and cerebellum in all patients. Binding potential (receptor concentration x affinity) was diffusely decreased in cerebral cortex, thalamus, striatum and cerebellum compared with control subjects, suggesting that GABAergic function may be decreased globally in these patients. Cerebral blood flow was largely normal, and no cerebral cortical atrophy was evident on MRI. CONCLUSION: Iodine-123-iomazenil SPECT may become a potent method for detecting impairment of the cerebral cortex even before brain perfusion SPECT or MRI can reveal early abnormalities. PMID- 9744337 TI - Pulmonary metastases in children and adolescents with well-differentiated thyroid cancer. AB - In this study, 27 patients less than 18 yr old with pulmonary metastases from well-differentiated thyroid carcinoma were evaluated to determine their response to (131)I therapy. METHODS: Of 121 children and adolescents treated with (131)I between 1963 and 1996, 27 patients had pulmonary metastases associated with nodal disease. Treatment response from (131)I was measured by three parameters: chest radiograph, scintigraphic images and serum thyroglobulin levels. Total activity of (131)I administered ranged from 4.6 GBq (125 mCi) to 38.7 GBq (1.05 Ci). Four patients were given one treatment, 8 were given two treatments, 4 were given three treatments and 11 were given more than three treatments. Radiation doses to the lungs were estimated in 14 patients using the MIRD methodology. The minimum duration of follow-up was 6 mo. RESULTS: At the time of initial presentation, diagnostic (131)I studies revealed bilateral radioiodine uptake in the lungs in 19 (70.4%) patients, whereas 12 (44.4%) patients had abnormal chest radiographs. One patient was lost to follow-up and was excluded from the study. Of the 26 patients studied, complete ablation of pulmonary metastases was observed in 8 (30.8%), partial ablation in 17 (65.4%) and there was no response to treatment in 1 (3.8%). Dosimetric parameters such as radioiodine uptake as a percentage of therapeutic activity, effective half-life and radiation dose delivered to the lungs were evaluated with each therapy. There was a progressive decline in each of these parameters with successive therapies. No correlation was observed between the radiation dose delivered and the response of pulmonary metastases to therapy. The number of therapies and amount of radioiodine administered had no influence on the ablation response. Of the 26 patients, 13 had a follow-up duration of less than 5 yr, 7 had 5-10 yr and 6 had more than 10 yr. One patient developed new metastases after 7 yr of diagnosis and treatment. One patient died of the disease after 4 yr. All surviving patients were asymptomatic and leading normal lives. CONCLUSION: Complete response of pulmonary metastases after (131)I therapy is difficult to achieve. A partial response with reduction of metastatic disease is possible and, in general, the patients had a good quality of life with no further disease progression and a low mortality rate. PMID- 9744335 TI - Dopamine transporter imaging with fluorine-18-FPCIT and PET. AB - Fluorinated N-3-fluoropropyl-2-beta-carboxymethoxy-3-beta-(4-iodophenyl) nortropane (FPCIT) has been synthesized as a dopamine transporter ligand for PET studies. We evaluated the regional brain uptake and the plasma metabolism of [18F]-FPCIT. METHODS: PET studies were conducted on 7 normal subjects and on 10 patients with Parkinson's disease. After the [18F]-FPCIT injection (4.4+/-1.8 mCi), dynamic scans were acquired over 100 min. Plasma metabolite analysis was performed using high-performance liquid chromatography (HPLC). RESULTS: Plasma HPLC revealed two peaks corresponding to unmetabolized [18F]-FPCIT and a polar metabolite. The fraction of the parent compound decreased rapidly to 25% at 25 min. Fluorine-18-FPCIT showed a striatum-to-occipital ratio (SOR) of 3.5 at 90 min postinjection. The ratio of striatal-to-occipital distribution volume (DVR) was calculated directly by using a mean tissue-to-plasma efflux constant for occipital cortex obtained in 10 subjects (ki=0.037 min(-1)). DVR measures determined with and without plasma input function were correlated (r=0.98, p < 0.0001). In normal subjects, a significant age-related decline of DVR was observed both for caudate and putamen, corresponding to a 7.7% and 6.4% decline per decade, respectively (r > 0.85, p < 0.01). Both DVR and SOR correctly classified early-stage Parkinson's disease patients with comparable accuracy (p < 0.0001). Age-corrected DVR values correlated negatively with the Uniform Parkinson's Disease Rating Scale composite motor ratings (r=0.66, p < 0.05). CONCLUSION: The tracer characteristics are compatible with a high-affinity, reversible ligand. FPCIT/PET demonstrated age-related decline in dopamine transporter binding in normal subjects as well as significant reductions in patients with idiopathic Parkinson's disease, which correlates with the disease severity. PMID- 9744338 TI - Technetium-99m-furifosmin in the follow-up of differentiated thyroid carcinoma. AB - Tumor scintigraphy with flow tracers, such as 201TI-chloride, has an established role in the follow-up of differentiated thyroid cancer. We investigated a new tracer, 99mTc-furifosmin (Technescan Q12, Mallinckrodt Diagnostika, Hennef, Germany), in patients with elevated thyroglobulin levels or sonographic suspicion of lymph node metastases or recurrent disease. METHODS: In a prospective study, we examined 20 patients with 99mTc-furifosmin. All patients underwent a 18F fluorodeoxyglucose (FDG) PET scan of the neck and chest. Positive 99mTc furifosmin findings were validated by biopsy, (131)I scan, CT or 18F-FDG PET examinations. RESULTS: In three patients with cervical lymph node metastases detected on a planar 99mTc-furifosmin scan, we found a rapid tracer accumulation in the tumor (maximum < 2 min) and a significant washout in 2 of 3 patients after 4 hr. The visual contrast and the tumor-to-nontumor ratio was rather poor (average 1.2:1). In 3 additional patients, 3 pulmonary and 2 mediastinal lymph node metastases were detected by the 99mTc-furifosmin SPECT scan. Two patients were true-negative, and in 13 of 18 patients, the tumor could be localized by 18F FDG PET (10 cervical, 6 mediastinal, 4 pulmonary metastases, 1 bone metastasis); 5 patients were false-negative. In 3 of these false-negative cases we could not localize the tumor with other diagnostic methods. Two patients had a true negative PET examination. CONCLUSION: The statistical analysis of our data on 99mTc-furifosmin reveals that the sensitivity and 95% confidence interval of 33% (11%-56%) on a patient-by-patient basis and of 34% (17%-57%) for the lesion-by lesion analysis is significantly lower than the sensitivity and 95% confidence interval of 72% (50%-89%) on a patient-by-patient basis and of 91% (78%-100%) on lesion-by-lesion basis for 18F-FDG. The sensitivity of 99mTc-furifosmin appears to be poor, even for cervical and mediastinal tumor manifestations where the value of 201TI-chloride is established. PMID- 9744339 TI - Role of initial iodine-131 whole-body scan and serum thyroglobulin in differentiated thyroid carcinoma metastases. AB - We evaluated the role of first (131)I-whole-body scan and of first serum thyroglobulin (Tg) measurement after surgery in the early diagnosis of metastases from differentiated thyroid carcinoma (DTC). METHODS: In 269 patients with metastases from DTC, we retrospectively evaluated the results of first whole-body scan (performed 40 days after surgery with diagnostic or therapeutic (131)I dose) and in 69 of them we also evaluated the result of first Tg measurement (performed the day before the first whole-body scan) in relation to the presence, localization and type of metastases. RESULTS: In all patients, the first whole body scan was positive for the thyroid remnant, and in 54.3% of patients it was also positive for metastases. In the remaining 45.7% of patients, metastases were detected during the follow-up. First Tg levels were >60 ng/ml in 66.7% of patients with metastases. First whole-body scan detected metastases in 47.8% of patients with Tg values <60 ng/ml, while Tg values were >60 ng/ml in 61.3% of patients with first whole-body scan negative for metastases. The combined results of both first whole-body scan and first Tg measurement allowed the early detection of metastases in 82.6% of patients. Whole-body scan detected distant metastases more frequently than local lymph node metastases (p < 0.01). CONCLUSION: In more than 80% of patients, metastases were suspected or diagnosed as early as 40 days after surgery in the presence of residual thyroid tissue by combined evaluation of results of first whole-body scan and Tg measurement. PMID- 9744341 TI - Intermediate and long-term side effects of high-dose radioiodine therapy for thyroid carcinoma. AB - The present investigation is an evaluation of intermediate and long-term side effects in patients after high-dose radioiodine treatment due to differentiated thyroid carcinoma. METHODS: A total of 203 patients were interviewed using a standardized questionnaire. RESULTS: After radioiodine treatment, 76.8% of the patients reported intermediate (from discharge up to 3 mo) or long-term (more than 3 mo after treatment) complaints, and 61.1% reported long-term side effects. Nonstochastic side effects included sialoadenitis, which occurred in 33.0% of cases, and 27.1% of patients suffered from a transient loss of taste or smell. More than 1 yr after the last radioiodine application, 42.9% of patients suffered from reduced salivary gland function. Complete xerostomia occurred in 4.4% of patients. Hematological abnormalities were found in 9 patients. In 28.1% of patients a transient episode of alopecia was reported. In 22.7% of patients chronic or recurrent conjunctivitis was reported, and 4 patients underwent dacryocystorhinostomy; 13.8% of patients suffered from an increased frequency of influenza, but 3.4% reported a reduced occurrence of such infections. For sialoadenitis, the loss of taste/smell and dry mouth, the dependence on accumulated activity was significant. CONCLUSION: Severe long-term side effects are rare after high-dose radioiodine treatment. Moderate side effects are common. The side effects are commonly the result of radiation damage to the salivary glands. The frequency of such complaints advocates regular protection of the salivary glands. PMID- 9744340 TI - Influence of scanning doses of iodine-131 on subsequent first ablative treatment outcome in patients operated on for differentiated thyroid carcinoma. AB - The therapeutic outcome after (131)I first ablative treatment in patients operated on for nonmedullary differentiated thyroid carcinoma was compared after both the currently used scanning dose of 111 MBq (131)I and a scanning dose of 37 MBq (131)I. METHODS: Two-hundred twenty-nine consecutive patients with no known metastases were retrospectively studied. They were divided in two populations according to the scanning dose (127 patients with 111 MBq and 102 patients with 37 MBq). All patients received 111 or 37 MBq (131)I for diagnostic purposes and 3.7 GBq (131)I for ablative therapy 9 days later. To assess the efficacy of the treatment, all patients were studied with (131)I and with thyroglobulin plasma assays 6-17 mo later. RESULTS: Successful outcome was significantly more frequent after a scanning dose of 37 MBq (131)I than after a scanning dose of 111 MBq (76% versus 50%, p < 0.001). The treatment efficacy was particularly enhanced after 37 MBq in patients with associated lymphocytic thyroiditis. CONCLUSION: In patients with no known metastases, our data suggest that the impairment of the treatment efficacy observed after a scanning dose of 111 MBq (131)I is related to a stunning effect on the thyroid remnants. The threshold amount above which this effect begins to occur in thyroid remnants could be between 37 and 111 MBq (131)I. Consequently, a scanning dose of only 37 MBq (131)I could be recommended before first ablative treatment. The absence of metastatic patients in our study prevents any conclusion about the possible stunning of the neoplastic tissue. Nevertheless, we must suspect such an effect and try to avoid it, especially during follow-up after first radioiodine therapy. For instance, one may consider postponing radioiodine treatment several weeks or even months after scanning dose administration or using only thyroglobulin measurement for patients who are likely to receive a subsequent radioiodine treatment. PMID- 9744342 TI - Redifferentiation therapy with retinoic acid in follicular thyroid cancer. AB - We report on a patient with a follicular Hurthle cell carcinoma in whom distant metastases were initially radioiodine negative or only weakly positive. Redifferentiation therapy with 13-cis retinoic acid induced a significant radioiodine uptake in metastatic tissue. Thyroglobulin (Tg) immunostaining and autoradiography of a bone metastasis in the right femur, which was initially radioiodine negative, proved Tg synthesis, combined with iodine incorporation into tumor cells. Glucose metabolism in metastases was partially increased and partially unchanged after redifferentiation therapy. The distinct increase of serum Tg after retinoic acid treatment was interpreted as a functional sign of redifferentiation. PMID- 9744343 TI - Remotely pollable Geiger-Muller detector for continuous monitoring of iodine-131 therapy patients. AB - In many countries, patients treated with therapeutic amounts of (131)I are hospitalized because of radiation safety considerations. To determine when they can return home, radiation levels are intermittently monitored at bedside using a handheld Geiger-MIller (GM) counter, although this procedure can be cumbersome and inexact. METHODS: We have developed and tested a remotely pollable system for continuous radiation monitoring of (131)I therapy inpatients, using readily available hardware and standard telephone lines. The remote detector system, consisting of a palmtop IBM-compatible personal computer, specialized software, PCMCIA modem and miniature serial port-based GM detector, is placed opposite the patient's bed at a fixed distance, and continuous 1-min acquisitions are started. Initially and at least twice daily, the remote palmtop is contacted by modem, and all interval data are uploaded onto the operator's base computer over the telephone line, including measurements taken with the patient in a predetermined standardized position. Continuous minute-to-minute data may be viewed in native form or can be imported into graphing and spreadsheet programs. Points acquired with the patient in standardized position are specially marked to highlight the constant geometry used. The ratio of initial counting rate to administered dose is used to estimate residual (131)I body burden by proportionality. Display of data as a semilogarithmic plot facilitates extrapolation of the activity curves and prediction of the patient's earliest time of discharge. RESULTS: We have characterized the remote GM detector system to confirm accuracy, counting rate linearity and reliability of data transfer. We describe examples that illustrate the applicability and usefulness of this method for remote monitoring of inpatient (131)I therapy levels. CONCLUSION: Monitoring patients with the described remotely pollable GM detector is an accurate and easy-to-implement technique that could conceivably lead to shortened hospital stays for (131)I therapy inpatients. Continuous quantitative data obtained are useful for kinetic and dosimetric analyses, which may be applied to study other gamma-emitting radiopharmaceuticals as well. The flexibility of the technique may permit its use in the monitoring of therapy on an outpatient basis, where allowed. PMID- 9744344 TI - Technetium-99m-MDP scintigraphy and long-term follow-up of treated primary malignant bone tumors. AB - Local malignant bone tumor excision followed by high-dose extracorporeal irradiation (300 Gy) and subsequent reimplantation is a unique technique for treatment of primary bone and cartilage tumors. The long-term scintigraphic findings of irradiated bone autografts in relation to clinical patient data were reviewed retrospectively. METHODS: Thirty-seven patients (12 women, 25 men; age range 13.0-66.7 yr; average age 29.1 yr) were studied. Postsurgical anatomopathological diagnoses included osteosarcoma, 20 patients; chondrosarcoma, 7 patients; and other less-frequent primary osteogenic tumors, 10 patients. Three hundred ninety 99mTc-methylene diphosphonate (MDP) whole-body scans performed between 3 mo and 18.3 yr (mean 6.5 yr) after treatment were reviewed. RESULTS: The 10-yr actuarial survival rate was 78%. After a mean period of 19.4 mo, 6 patients developed a local recurrence, and MDP scintigraphy detected the recurrence in 4. Distant metastases developed in 11 patients (30%), of which 10 were nonosseous. Initially, all autografts appeared as photon-deficient areas. Diffusely increased bone uptake was present at osteotomy sites and at articulating surfaces contiguous with autografts within the first few months after surgery. Of all 25 patients with adequate follow-up, 7 showed persistent decreased uptake up to 129 mo after surgery. The other patients developed partial tracer uptake after 19.6 mo, on average. In 6 patients, scintigraphic images consistent with complete revascularisation were noted later (mean 31.5 mo). Local, sometimes multiple, complications were noted in 22 patients, mainly mechanical graft-related (15) or infections (11). Scintigraphic sensitivity for mechanical complications was 100%. Significantly more fractures and collapses were seen when partial tracer uptake suggestive of revascularisation occurred. Altered bone stress gave rise to focal and diffuse scintigraphic abnormalities, often in the spine and lower extremities. In recent literature, similar clinical complication patterns are found for massive allografts. CONCLUSION: Skeletal scintigraphy is a sensitive technique for evaluating long-term follow-up of massive grafts to treat primary malignant bone tumors. Revascularisation and partial bone ingrowth are not sufficient conditions for a lower complication rate. PMID- 9744345 TI - Iodine-123-vasoactive intestinal peptide receptor scanning in patients with pancreatic cancer. AB - Recent data demonstrated a high sensitivity (>90%) in the visualization of primary/recurrent pancreatic cancer as well as metastases by means of 123I labeled vasoactive intestinal peptide (VIP). The aim of this study was to investigate the diagnostic value of radioiodinated VIP in patients suffering from adenocarcinoma of the exocrine pancreas. METHODS: Sixty consecutive patients (26 women, 34 men; mean age 59 yr) with histologically verified pancreatic cancer were investigated in this study. Twenty-one patients presented with organ confined malignancy (19 at study entry and 2 during follow-up after initial surgery developed tumor recurrence), while 25 patients had distant metastases along with the local malignancy, and 7 patients had liver metastases after resection of the primary lesion (6 on study entry and 1 during follow-up showed tumor development). In 5 of these patients, abdominal lymph node metastases were present at the time of scanning. Of 10 patients, who had undergone potentially curative surgery for their cancer, 7 remained free of disease during follow-up until death or for at least 6 mo. Iodine-123-VIP (150-200 MBq; approximately 1 microg VIP) was administered to all patients. Scintigraphic results were evaluated as compared to conventional radiologic imaging methods and surgical exploration. RESULTS: Primary pancreatic tumors were visualized by 123I-VIP in 19/21 patients (90%) with disease confined to the pancreas and in 8/25 patients (32%) suffering both from locoregional and disease metastatic to the liver. The overall 123I-VIP scan sensitivity for primary pancreatic adenocarcinomas was 58% (27/46 scans). Liver metastases were imaged in 29/32 patients (scan sensitivity 90%) and abdominal lymph node metastases in 4/5 patients. In 5 patients, the VIP receptor scan indicated the malignant lesion before CT. In vitro results confirmed specific binding of 123I-VIP to primary pancreatic tumor cells as well as to PANC1 adenocarcinoma cells. CONCLUSION: Iodine-123-VIP receptor scanning has the potential to offer additional information to augment diagnostic standard methods and could influence the decision-making process in the treatment of pancreatic cancer. PMID- 9744346 TI - Location of a VIPoma by iodine-123-vasoactive intestinal peptide scintigraphy. AB - A major problem in patients with small endocrine tumors is the difficulty in localizing the primary tumor site. Many endocrine tumors possess larger amounts of high affinity vasoactive intestinal peptide (VIP) binding sites compared with normal tissue or blood cells. We used radiolabeled VIP to localize the tumor site in a patient with Verner-Morrison syndrome (VMS). Under octreotide therapy, the VIP levels had declined in this patient, but a tumor site could not be detected by conventional techniques or by radiolabeled octreotide. However, using 123I VIP, the tumor was detectable in the pancreatic tail. Surgical resection of the tumor was followed by complete remission of the VMS. Expression of VIP binding sites in the tumor was confirmed by a radioreceptor assay and showed cross competition between VIP and octreotide. The identity of the VIP binding site in the tumor was analyzed by Northern blotting and revealed the expression of somatostatin receptor subtype 3, which binds both somatostatin-14 and VIP with higher affinity than octreotide. Iodine-123-VIP scintigraphy would be an effective tracer to identity the tumor site in VMS patients. PMID- 9744347 TI - Indium-111-DTPA-folate as a potential folate-receptor-targeted radiopharmaceutical. AB - Indium-111-labeled diethylenetriamine pentaacetic acid (DTPA)-folate was evaluated as a radiopharmaceutical for targeting tumor-associated folate receptors. METHODS: Athymic mice were subcutaneously inoculated with approximately 1.8 x 10(6) folate receptor-positive KB (human nasopharyngeal carcinoma) cells, yielding 0.2- to 0.6-g tumors in 15 days, at which time (111)In DTPA-folate, (111)In-DTPA or (111)In-citrate was administered by intravenous injection. RESULTS: The (111)In-DTPA-folate conjugate afforded marked tumor specific (111)In deposition in vivo using this mouse model. The involvement of the folate receptor in mediating tumor uptake of (111)In-DTPA-folate was demonstrated by the blocking of tumor uptake by coadministration of free folic acid (intravenous). The (111)In-DTPA-folate also shows folate receptor-mediated uptake and retention in the kidneys, presumably reflecting radiotracer binding to folate receptors of the proximal tubules. In control experiments, the (111)In citrate radiopharmaceutical precursor was also shown to afford significant tumor uptake of (111)In, but with much poorer tumor-to-background tissue contrast than that obtained with (111)In-DTPA-folate. Unconjugated (111)In-DTPA showed no tumor affinity. CONCLUSION: Indium-111-DTPA-folate appears suitable as a radiopharmaceutical for targeting tumor-associated folate receptors. PMID- 9744348 TI - Gallium-67 scintigraphy evaluation of therapy in non-Hodgkin's lymphoma. AB - Patients with diffuse large cell lymphoma may achieve complete remission (CR) after chemotherapy, and the time to reach CR may be predictive of treatment outcome. Partial remission, or recurrence from CR, is associated with poor survival. Gallium-67 imaging has proven to be useful in evaluating lymphoma patients. In tumor models, this radiotracer is an indicator of tumor viability. Gallium-67 uptake is seen only in avid and viable lymphoma tissue, not in fibrotic or necrotic tissue. In this study, we prospectively assessed the ability of this radiotracer to define residual disease. In addition, we evaluated the possibility of predicting the clinical outcome in patients with diffuse cell lymphoma on the basis of scan positivity during chemotherapy. METHODS: Thirty three consecutive patients with histologically proven diffuse large cell lymphoma were investigated with 67Ga scintigraphy 48-72 hr after injection of 185-259 MBq 67Ga-citrate for staging and during follow-up after four to six cycles of intensive chemotherapy. Patients were monitored for a mean of 56.0 mo (range 7-90 mo), and they were restaged using physical examination, CT and all necessary imaging modalities. RESULTS: Patients were divided into two groups according to the positivity or negativity of 67Ga scan after four to six cycles of chemotherapy. Of the 33 patients studied, 14 (42.4%) showed persistent abnormal uptake of 67Ga-citrate after four to six cycles of chemotherapy. In this group, 9 patients (64.2%) died of lymphoma at a mean of 24.3 mo from presentation with the diagnosis (range 7-71 mo). Four patients had no evidence of disease at an average of 71.7 mo after diagnosis, and 1 patient was considered to be in partial remission. In the second group of 19 67Ga-negative patients, after four to six cycles of chemotherapy, 4 died and 15 are alive and considered to be in CR. A statistical analysis of the association between 67Ga scan results after four to six cycles of chemotherapy and survival was performed using the log-rank test; there was a statistically significant association between scan results and survival (p=0.00125). CONCLUSION: We conclude that 67Ga scintigraphy is an excellent predictor of residual tumor viability in lymphoma patients and that persistent positivity of the scan predicts poor outcome and may justify a change in treatment. PMID- 9744349 TI - Impact of scatter correction in planar scintimammography: a phantom study. AB - This study examines how scatter correction might affect lesion detection and quantitation of tumor-to-normal breast tissue activity ratio in planar scintimammography. METHODS: Forty-one phantom acquisitions were performed to mimic a wide variety of scintimammographic imaging conditions in which lesions would be close to the chest wall. For each acquisition, the images corresponding to a 10% energy window (110) and two scatter correction methods [the Jaszczak (JA) method and a factor analysis (FA)-based method] were obtained in addition to the conventional 20% image (120). A total of 368 images in which detection of the "tumor" was judged borderline were selected, and 10 independent observers were asked to detect lesions in these images. Receiver operating curve analyses were performed to assess detection performance. Tumor-to-normal tissue activity ratios were calculated for quantitative analysis. RESULTS: Detection performance significantly improved for the I10, JA and FA images compared to the 120 images, with an increase in sensitivity up to 8% for FA images. Sensitivity was especially increased for small lesions (13- and 16-mm3 spheres) and true heart-to normal tissue activity ratios of > 12. Scatter correction also increased the certainty with which the readers gave their judgment. The tumor-to-normal tissue activity ratio was approximately 8% larger on JA or FA images and 1% larger on the I10 images compared to the 120 images. For a given image, the variability with which this ratio was estimated was reduced by approximately 4% on JA and FA images. CONCLUSION: Based on these phantom results, scatter correction might be used with benefit in scintimammography. PMID- 9744350 TI - Comparison of iodotyrosines and methionine uptake in a rat glioma model. AB - This study compares brain tumor imaging with 3-[123I]iodo-alpha-methyl-L-tyrosine (IMT) and 3-[123I/125I]iodo-O-methyl-alpha-methyl-L-tyrosine (OMIMT) to that with [methyl-3H]-L-methionine (Met) in a rat glioma model by double-tracer autoradiography. METHODS: Cells of the glioma clone F-98 were implanted stereotactically into the right basal ganglia of 22 Fischer 344 rats. After 8 days of tumor growth, the animals simultaneously were injected with a mixture of either 123I-IMT and 3H-Met (n=5), 123I-OMIMT and 3H-Met (n=8) or 123I-IMT and 125I-OMIMT (n=9). The animals were killed 15 min after the tracer injection and cryosections of the tumor-bearing brain area were exposed to phosphor-imaging plates both immediately and after the decay of 123I. Tumor-to-brain ratios (T/B) and intratumoral distribution of the different tracers and of the cresyl violet staining of the tissue were compared. RESULTS: There was a significant correlation of the T/B ratios between all tracers (IMT versus Met: r=0.97, n=5, p < 0.01; OMIMT versus Met: r=0.94, n=8, p < 0.001; OMIMT versus IMT: r=0.95, n=9, p < 0.001). Intratumoral tracer distribution was similar for all tracers and the extent of tumor labeling was identical to that of the histological tumor extent. Mean values of the T/B ratios, however, were lower for IMT (2.81+/-0.78, n=14, mean+/-s.d., p < 0.01 compared with Met) and for OMIMT (2.03+/-0.57, n=17, p < 0.01 compared with Met) than for Met (3.86+/-1.12, n=13). CONCLUSION: This study confirms that tumor imaging with IMT is similar to that of Met but T/B ratios of IMT are lower. OMIMT intratumoral tracer distribution and tumor size are similar to Met and IMT, but the T/B contrast is rather low and makes this amino acid less suitable for clinical application. PMID- 9744352 TI - Intense muscle uptake of gallium-67 in a patient with sarcoidosis. AB - Sarcoidosis has been associated with muscle involvement. In general, this involvement remains asymptomatic. The following case report demonstrates a patient with a 4-mo history of sarcoidosis who reported severe fatigue and slight muscular complaints at a regular checkup. Gallium scintigraphy indicated unexpected and unusually extensive muscular localizations of the disease. The latter findings were confirmed by examination of biopsy specimens. The importance of gallium scintigraphy lies in the possibility of wholebody screening for inflammation localizations, particularly when physical, laboratory, lung function and radiographic examinations fail to provide convincing evidence of active sarcoidosis. Furthermore, it can be helpful in the follow-up of the effect of supportive treatment. PMID- 9744351 TI - Midcourse thallium-201 scintigraphy to predict tumor response in bone and soft tissue tumors. AB - The purpose of this study was to assess the predictive power of 201TI scintigraphy in the midcourse of chemotherapy for the final tumor response to chemotherapy in malignant bone and soft-tissue tumors. METHODS: The 21 patients studied with 201TI scintigraphy were 14 males and 7 females (average age 39.8+/ 22.1 yr; age range 8-74 yr). Planar scintigraphy was performed 15 min after injection of 111 MBq 201TI before chemotherapy, after the third chemotherapy cycle (midcourse) in all 21 patients and after the final chemotherapy cycle but before surgery in 11 patients. The 201TI uptake ratio was calculated by dividing the count density of the lesion by that of the contralateral normal area. The percent reduction of the 201TI uptake ratio calculated by 100 x [(prechemotherapy ratio - postchemotherapy ratio)/prechemotherapy ratio] in the midcourse was compared with that after the final course of chemotherapy, and it also was compared with the histologic response. RESULTS: In patients with histologically complete response [(CR), n=6] and with partial response [(PR), n=5], the percent reduction in 201TI uptake ratio after three cycles of chemotherapy was 64.1%+/ 14.4% and 50.9% +/-10.5%, respectively. In patients with histologically no change [(NC), n=10], the percent reduction was 0.40%+/-18.2% after the third cycle; 5.3%+/-20.9% in four patients with full courses of chemotherapy (p < 0.0001 and p < 0.005 compared with the CR and PR groups, respectively). After the final cycle of chemotherapy, the percent reduction in 201TI uptake ratio was 68.6%+/-14.7%, 56.2%+/-6.1% and -0.3%+/-17.2% in the CR, PR and NC groups, respectively (NC versus CR, p < 0.0005; NC versus PR, p < 0.005). CONCLUSION: Thallium-201 scintigraphy performed in the midcourse of chemotherapy is predictive of the final response to chemotherapy that can be demonstrated histologically. Serial 201TI scintigraphy in the midcourse of chemotherapy is useful in assessing final chemotherapeutic response in the early stage of chemotherapy, and it helps clinicians when choosing the most appropriate treatment strategies in patients with bone and soft-tissue tumors. PMID- 9744353 TI - Biodistribution and dosimetric study in medullary thyroid cancer xenograft using bispecific antibody and iodine-125-labeled bivalent hapten. AB - The purpose of this study was to evaluate biodistributions and absorbed doses of anti-carcinoembryonic antigen (CEA)/anti-diethylenetriamine pentaacetic acid (DTPA)-indium (anti-DTPA-In) bispecific monoclonal antibody (BsMAb) F6-734 and 125I-labeled DTPA-indium dimer hapten (125I-di-DTPA-In hapten) in athymic mice xenografted with human medullary thyroid cancer. METHODS: Bispecific monoclonal antibodies F6-679 (anti-CEA/antihistamine) and G7A5-734 (antimelanoma/anti-di DTPA-In) were used as irrelevant BsMAbs. Athymic mice inoculated with TT medullary thyroid cancer cells expressing CEA were administered BsMAbs F6-734, F6 679 or G7A5-734 and then, 48 hr later, 125I-di-DTPA-In hapten. Iodine-125-labeled F6 F(ab')2 fragment was injected into other groups of mice. Biodistributions were examined at 30 min and 5, 24, 48 and 96 hr after injection of 125I-di-DTPA-In hapten or 125I-labeled F6 F(ab')2. RESULTS: In mice injected with BsMAb F6-734 and 125I-di-DTPA-In hapten, tumor uptake was 9.1%+/-2.1%, 8.7%+/-3.5%, 8.0%+/ 2.3%, 5.1%+/-0.9% and 3.5%+/-1.5% of the injected dose/g at 30 min and 5, 24, 48 and 96 hr, and tumor-to-blood, tumor-to-liver and tumor-to-kidney ratios were 37.0+/-12.5, 32.3+/-10.9 and 10.4+/-2.7 at 24 hr. Iodine-125-F6 F(ab')2 fragment showed a tumor uptake of 7.39% injected dose/g and tumor-to-blood, tumor-to-liver and tumor-to-kidney ratios of 1.8+/-0.6, 7.3+/-2.9 and 3.6+/-1.6 at 24 hr. In mice injected with F6-679 or G7A5-734, tumor uptake and tumor-to-normal tissue ratios were much lower than in the mice injected with F6-734. These results were confirmed by autoradiographic studies that demonstrated clear tumor-to-normal tissue contrast. CONCLUSION: This two-step targeting method seems very potent for the diagnosis and therapy of human medullary thyroid cancer and other CEA producing tumors because it combines high tumor uptake and low normal tissue background. PMID- 9744354 TI - Prediction of bone loss in patients with primary hyperparathyroidism using quantitative bone SPECT. AB - Bone loss is a major complication of primary hyperparathyroidism (PHPT), and it has significant implications in the treatment of this disease. Bone turnover was measured in patients with PHPT, using quantitative bone SPECT (QBS), to determine if the rate of bone loss could be predicted before a significant decrease in bone mass occurs. METHODS: Forty-six patients were included in the study. QBS and bone mineral density (BMD) of the lumbar spine (LS) and femoral neck (FN) were done at baseline. The percent deviation of QBS in patients with PHPT from the values in normal matched controls was calculated. BMD was measured again after a mean of 17.5 mo in 38 patients, and in 29 patients a repeat BMD study was done after a mean of 41.4 mo. The change in BMD in patients with high and normal QBS values was compared using the nonparametric Mann-Whitney test. Regression analysis tested the correlation between baseline QBS values and BMD changes over time. RESULTS: For the FN, there was a statistically significant difference in the BMD change between patients with high and normal QBS values for short-term follow-up (-2.82%+/-4.80% versus 1.45%+/-4.67%, p < 0.05) and for long-term follow-up ( 3.53%+/-5.34% versus 0.92%+/-2.40, p < 0.02). There was a negative correlation in the FN, r=-0.48 between QBS values and the percentage of change in BMD. There was no significant difference between the percentage of change in BMD in the LS in patients with high and normal QBS values for either short- or long-term follow up. CONCLUSION: The results of this study show that QBS can predict bone loss in the FN in patients with PHPT. QBS can thus indicate the need for surgery at an early stage of the disease to prevent bone loss. PMID- 9744355 TI - Technetium-99m-sestamibi parathyroid scintigraphy: effect of P-glycoprotein, histology and tumor size on detectability. AB - The purpose of this study was to evaluate the effect of P-glycoprotein (P-gp) levels, predominant histology and tumor size on the detectability of parathyroid adenomas with 99mTc-sestamibi scans. Although previous studies have shown that positivity correlates with tumor size, false-negative studies have been reported with large tumors and true-positives reported with very small tumors. Recent investigations have reported rapid washout of sestamibi in malignant tumors because of high levels of P-gp, similar to that seen with multidrug chemotherapy resistance. Therefore, we postulated that this mechanism might account for false negative studies in parathyroid tumors. Preliminary reports have suggested that the predominant histological subtype influences positivity on 99mTc-sestamibi parathyroid scans. METHODS: We studied 25 patients with surgically confirmed parathyroid adenomas with 99mTc-sestamibi parathyroid scans. The results of the imaging study were correlated with tissue P-gp levels, predominant histological subtype and size of the surgically removed glands. RESULTS: There were 21 true positive and 4 false-negative results. The size of the adenomas ranged from 0.12 to 8.64 ml. We found no correlation between the results of the dual-phase 99mTc sestamibi study and either the predominant cell type or the level of P-gp. Positivity did correlate with the size of the adenoma (p=0.73, p < 0.0001). We cannot exclude the possibility that P-gp and cell type may still play a role in individual cases, but we suspect that other yet to be determined factors may influence 99mTc-sestamibi detectability in addition to tumor size. PMID- 9744356 TI - False-positive captopril renography in patients taking calcium antagonists. AB - Captopril-enhanced renography is the noninvasive test of choice for the diagnosis of renovascular hypertension. Previous studies have shown that bilateral symmetrical changes are associated with many renal conditions. However, patients with normal renal angiography occurred in our institutions despite this scintigraphic pattern, and no known conditions could explain these results. The purpose of this study was to evaluate the diagnostic implications of bilateral symmetrical renal function deterioration on captopril renography. METHODS: Eighty six captopril renal scintigraphies performed at two centers to exclude renovascular hypertension (50 consecutive patients after the observation of a bilateral symmetrical renal function deterioration despite a normal angiogram at one institution and 36 patients with both captopril renography and renal angiography at the other institution) were retrospectively reviewed. Baseline and captopril-enhanced renograms were obtained with 99mTc-mercaptoacetyltriglycine and a 1-day protocol in 50 patients; 36 patients were studied using 99mTc diethylenetriamine pentaacetic acid and a 2-day protocol. Bilateral symmetrical renal function deterioration was detected. RESULTS: Ten patients presented with bilateral symmetrical renal function deterioration on their captopril renograms; 9 of them were taking calcium antagonists (p=0.015). Control studies performed in 5 patients without these medications demonstrated normal captopril renograms in 4 and persistent renal dysfunction in 1. No explanation was found for the patient who was not taking any medication. Angiograms performed in 5 patients showed normal renal arteries. An 11th patient who was taking a calcium antagonist showed dysfunction of his one kidney on the captopril renogram but no artery stenosis on the renal angiogram. CONCLUSION: Calcium antagonists can cause false-positive captopril renograms. These medications should be stopped before captopril renography, and physicians should be aware of this possible drug interaction if bilateral symmetrical renal function deterioration is seen on a patient's captopril renogram. PMID- 9744357 TI - Comparative study of technetium-99m-sestamibi and thallium-201 SPECT in predicting chemotherapeutic response in small cell lung cancer. AB - The purpose of this study was to evaluate the relationship between 99mTc sestamibi (MIBI) accumulation by tumor and response to chemotherapy in small cell lung cancer patients compared with 201TI-chloride. METHODS: There were 19 patients with small cell lung cancer just before chemotherapy initiation. The patients were classified by a follow-up CT into complete remission, partial remission and no change groups. All patients underwent dual-isotope imaging with 201TI-chloride and 99mTc-MIBI. Regions of interest were placed over the tumors and contralateral normal lung tissue on one coronal view with a clearly defined lesion, and the tumor-to-normal (T/N) ratio and retention index were calculated. RESULTS: Early and delayed T/N ratios for 99mTc-MIBI in complete remission and partial remission groups were significantly higher (p < 0.05) than in the no change group. There was no significant correlation between T/N ratio and tumor response using 201TI-chloride. There were no significant differences in the retention index with respect to the tumor response in both 201TI-chloride and 99mTc-MIBI SPECT images. CONCLUSION: Technetium-99m-MIBI SPECT may be more effective than 201TI-chloride SPECT for evaluating response to chemotherapy in patients with small cell lung cancer. PMID- 9744358 TI - Quantitative analysis of nonuniform distributions in lung perfusion scintigraphy. AB - Nonuniform distributions of lung perfusion scintiscans obtained by SPECT were quantified to supplement qualitative lung scintiscans. A total of 126 lung perfusion scintigraphy examinations were performed between February and December 1996 on a subject population of 102 patients, including 8 control subjects. All of the subjects were broadly classified according to whether they had pulmonary disease or nonpulmonary disease. The latter group was subdivided into a group with cardiac disease and other diseases. The grade of breathlessness was classified according to whether oxygen inhalation was required, the clinical severity of the breathlessness and patient's performance status. Blood gas analysis was performed in 21 cases, and pulmonary function testing was performed in 26 cases. METHODS: With the subjects resting in the supine position, 185 MBq 99mTc-macroaggregate albumin was infused. From reconstructed SPECT images, the volume of lung as a whole calculated at 10% of threshold was assumed to be the functional lung volume, and the functional volume rates were calculated in 10% threshold widths from 10% to 100% of thresholds. Assuming the total absolute difference in functional volume rate between each subject and the control to be the distribution index of the lung as a whole (D index), we quantified the degree of nonuniform distribution in each subject. RESULTS: The D index of all subjects ranged from 2.7 to 72.2. The mean D index in pulmonary disease was significantly higher than in nonpulmonary disease (p < 0.0005) and cardiac disease (p < 0.005). It was significantly positively correlated with the grade of breathlessness, significantly negatively correlated with the oxygenation index and significantly positively correlated with measured vital capacity and forced expiratory volume in 1 sec as percentages of their predicted values. CONCLUSION: The D index is a useful indicator for quantifying nonuniform distributions on lung scintiscans. If it is used as a supplement to qualitative interpretation of scintiscans, pulmonary perfusion scintigraphy will become a more useful technique for clinical evaluation of treatment and assessment of breathlessness and respiratory failure than the usual one. PMID- 9744359 TI - Lymphoscintigraphy and lymphangiography of lymphangiectasia. AB - Chronic genital edema secondary to lymphangiectasia and chylous reflux in a 23-yr old man with Noonan syndrome was investigated by 99mTc sulfur nanocolloid lymphoscintigraphy and bipedal contrast lymphangiography. Lymphoscintigraphy showed a delayed lymphatic flow pattern in the pelvis, abdomen and chest consistent with lymphangiectasia and abnormal lymphatic flow dynamics. Lymphangiography showed dilated and tortuous abnormal lymphatics in the abdomen and pelvis. Ligation of incompetent retroperitoneal lymph vessels and lymphaticovenous anastamosis were performed, resulting in clinical improvement. Lymphangiectasia has been described previously in Noonan syndrome, but it is relatively uncommon below the diaphragm. This case demonstrates the use of lymphoscintigraphy and lymphangiography in providing important physiological and anatomical information before surgical intervention. Careful presurgical planning using such tests also allows the most appropriate operation to be performed. PMID- 9744360 TI - Rat antigen-induced arthritis: cartilage alterations assessed with iodine-123 antileukoproteinase. AB - Imaging of cartilage alterations was attempted in joints of rats with chronic antigen-induced arthritis (AIA) using the cationic 123I-labeled serine proteinase inhibitor antileukoproteinase (123I-ALP; pI > 10), which selectively accumulates in normal cartilage, presumably through interaction with negatively charged proteoglycans. METHODS: Iodine-123-ALP or 123I-myoglobin, a control protein of comparable size but with different isoelectric point (pI=7.3) was injected intravenously into normal or AIA rats. Joint accumulation was followed by scintigraphy for 14 hr. Tissue radioactivity was assessed by well-counter measurements after dissection. The content of charged molecules in articular cartilage was determined by toluidine blue staining; the degree of joint destruction was assessed in parallel by x-ray, ex vivo MRI and histopathology. RESULTS: In intact articular cartilage, ALP accumulated to a significantly higher degree than myoglobin. This preferential accumulation was lost in rats with chronic AIA. The target-to-background ratio for 123I-ALP negatively correlated with the loss of toluidine blue staining in cartilage, which documents depletion of charged matrix molecules (r=-0.92, p < 0.01 at 4 hr; r=-0.97, p < 0.01 at 13 hr). ALP scintigraphy was sensitive in detecting cartilage alterations, even though the degree of joint destruction and inflammatory infiltration was mild, as demonstrated by x-ray, MRI and histopathology. CONCLUSION: In rat AIA, loss of ALP accumulation appears to document proteoglycan depletion in mildly altered arthritic cartilage. ALP scintigraphy may represent a functional assay for early, premorphological cartilage alterations in human arthritis as well. PMID- 9744361 TI - Estimating glomerular filtration rate in children. PMID- 9744362 TI - Physical and chemical nature of technegas. PMID- 9744363 TI - Interactive compartmental modeling. PMID- 9744364 TI - Is it time for a change? PMID- 9744366 TI - Relationship of the actual thick intraocular lens optic to the thin lens equivalent. AB - PURPOSE: To theoretically derive and empirically validate the relationship between the actual thick intraocular lens and the thin lens equivalent. METHODS: Included in the study were 12 consecutive adult patients ranging in age from 54 to 84 years (mean +/- SD, 73.5 +/- 9.4 years) with best-corrected visual acuity better than 20/40 in each eye. Each patient had bilateral intraocular lens implants of the same style, placed in the same location (bag or sulcus) by the same surgeon. Preoperatively, axial length, keratometry, refraction, and vertex distance were measured. Postoperatively, keratometry, refraction, vertex distance, and the distance from the vertex of the cornea to the anterior vertex of the intraocular lens (AV(PC1)) were measured. Alternatively, the distance (AV(PC1)) was then back-calculated from the vergence formula used for intraocular lens power calculations. RESULTS: The average (+/-SD) of the absolute difference in the two methods was 0.23 +/- 0.18 mm, which would translate to approximately 0.46 diopters. There was no statistical difference between the measured and calculated values; the Pearson product-moment correlation coefficient from linear regression was 0.85 (r2 = .72, F = 56). The average intereye difference was 0.030 mm (SD, 0.141 mm; SEM, 0.043 mm) using the measurement method and +0.124 mm (SD, 0.412 mm; SEM, 0.124 mm) using the calculation method. CONCLUSION: The relationship between the actual thick intraocular lens and the thin lens equivalent has been determined theoretically and demonstrated empirically. This validation provides the manufacturer and surgeon additional confidence and utility for lens constants used in intraocular lens power calculations. PMID- 9744367 TI - Chorioretinal damage caused by the excision of choroidal neovascularization. AB - PURPOSE: To determine whether choroidal neovascularization excision causes mechanical damage to the neurosensory retina, retinal pigment epithelium, or choriocapillaris. METHODS: Prospectively, 18 eyes of 18 consecutive patients who underwent choroidal neovascularization excision were observed. Preoperatively and postoperatively, the integrity of the choriocapillaris circulation in the pathway of choroidal neovascularization extraction was studied by fluorescein and indocyanine green angiography. Using static scanning laser ophthalmoscope microperimetry, the presence of iatrogenic scotomas that developed postoperatively in the pathway of choroidal neovascularization extraction was also investigated. RESULTS: Postoperatively, a choriocapillaris defect was detected in 17 (94.4%) of 18 cases. In 15 cases (83.3%), the choriocapillaris defect had a clear relationship to the pathway of choroidal neovascularization extraction. Postoperatively, a scotoma was present in 16 (88.9%) of 18 cases. In 14 cases (77.8%), the location of the scotoma had a clear relationship to the pathway of choroidal neovascularization extraction. CONCLUSION: Surgical excision of choroidal neovascularization leads to severe damage of the choroid and retina in the pathway of the extracted choroidal neovascularization. The injury involves the neurosensory retina, retinal pigment epithelium, and choriocapillaris. PMID- 9744368 TI - Management of submacular hemorrhage associated with retinal arterial macroaneurysms. AB - PURPOSE: Experience is reported with intraoperative pharmacologic lysis of recent submacular hemorrhage with tissue plasminogen activator followed by surgical drainage of the unclotted blood in patients with retinal arterial macroaneurysms. METHODS: Nine eyes (nine patients) with a recent (< or = 7 days old) submacular hemorrhage involving the center of the fovea secondary to retinal arterial macroaneurysm that were managed with recombinant tissue plasminogen activator assisted subretinal hemorrhage evacuation, including subretinal injection of tissue plasminogen activator and removal of the liquefied blood. Patients were followed for a mean 18 +/- 7 months (range, 7 to 30 months). RESULTS: All nine eyes had improved final corrected visual acuity after surgery, and eight eyes (89%) attained a corrected visual acuity of 20/60 or better (mean, 20/40; range, 20/20 to 20/200). Final corrected visual acuity was limited to 20/200 in one eye. Two eyes developed a cataract that required surgery. CONCLUSIONS: Submacular surgery with tissue plasminogen activator-assisted thrombolysis achieved improved best-corrected visual acuity in eyes with recent submacular hemorrhage involving the center of the fovea associated with retinal arterial macroaneurysm. PMID- 9744369 TI - The Collaborative Ocular Melanoma Study (COMS) randomized trial of pre enucleation radiation of large choroidal melanoma III: local complications and observations following enucleation COMS report no. 11. AB - PURPOSE: To summarize local complications and observations following enucleation of eyes with large choroidal melanoma that were reported prospectively at scheduled examinations of patients enrolled in the Collaborative Ocular Melanoma Study (COMS) randomized trial of pre-enucleation radiation. METHODS: Of 1,003 patients with large choroidal melanoma who were assigned randomly at time of enrollment to enucleation alone or to pre-enucleation radiation, 994 were treated as assigned. Complications and observations were reported to the Coordinating Center on standard forms completed at the time of enucleation surgery and during the immediate 24-hour postsurgery period, 1 to 6 weeks following surgery, 6 and 12 months after enucleation, and at examinations scheduled at 12-month intervals thereafter. RESULTS: The most common perioperative complication was pain that prolonged hospital stay, which was reported for six patients (1%) who had standard enucleation and eight patients (2%) who had pre-enucleation radiation. Patients treated with pre-enucleation radiation had somewhat more complications reported at the examination 1 to 6 weeks after surgery than did patients treated with enucleation alone, 36 (8%) and 21 (4%), respectively (P = .03, chi2 test), but all complications were minor. During follow-up, fewer biopsy-confirmed tumor recurrences in the orbit were observed among patients treated with pre enucleation radiation than with enucleation alone (0 vs 5, respectively; P = .03, Fisher exact test). Patients treated with pre-enucleation radiation also had a lower incidence of severe ptosis than did patients treated with enucleation alone (P = .007, log rank test). Among 307 patients examined 5 years after enucleation, the most frequent complication reported at the 5-year examination was poor prosthetic motility for 24 patients (16%) treated with enucleation alone and 30 patients (19%) treated with pre-enucleation radiation. CONCLUSIONS: Complications were infrequent during the 5-year period following enucleation surgery. Five-year incidence rates and prevalence at the 5-year examination of most complications were similar in the two treatment arms. There was no indication that pre enucleation radiation had resulted in more serious complications. PMID- 9744370 TI - Distribution of prognostically important vascular patterns across multiple levels in ciliary body and choroidal melanomas. AB - PURPOSE: To investigate the validity of assigning patients whose eyes have been removed for ciliary body or choroidal melanoma to risk groups for metastasis based on the identification of microcirculatory patterns in one cross-section taken from the center of the tumor. METHODS: Multiple levels were cut through the blocks of 15 ciliary body or choroidal melanomas until the tumor was exhausted. Each level was examined for the presence of microvascular networks and parallel vessels with cross-linking histologic features strongly associated with death from metastatic melanoma. RESULTS: The central histologic section did not contain either microvascular networks or parallel vessels with cross-linking in eight tumors, nor were these patterns encountered in any of the more peripheral levels of the tumor. Seven tumors contained at least one focus of either microvascular networks or parallel vessels with cross-linking in the central histologic section. In two tumors, at least one of these patterns appeared in all histologic levels; in five tumors, at least one of these patterns appeared through multiple levels until just before the tumor was exhausted from the block (0.24 to 0.85 mm from the edge of the tumor). CONCLUSIONS: This study suggests that the prognostic classification of uveal melanoma based on the histologic profile of the microcirculation may be consistent throughout the tumor depth. PMID- 9744371 TI - Neovascular complications associated with rubeosis iridis and peripheral retinal detachment after retinal detachment surgery. AB - PURPOSE: To report clinical features and surgical management of neovascular complications associated with rubeosis iridis and peripheral retinal detachment after retinal detachment surgery in nondiabetic patients. METHODS: Seven consecutive eyes of seven nondiabetic patients who developed neovascular complications associated with rubeosis iridis and peripheral retinal detachment after scleral buckling and vitrectomy procedures were retrospectively reviewed. None of the eyes had clinical evidence of anterior segment ischemia or retinal vascular disease, but each eye developed rubeosis iridis and neovascular complications. RESULTS: Of the seven eyes with rubeosis iridis and peripheral retinal detachment, six developed recurrent or progressive vitreous hemorrhage, and three developed progressive neovascular glaucoma. Four eyes underwent a revision procedure to repair the peripheral retinal detachment, and anterior proliferative vitreoretinopathy was found in each of these cases. Rubeosis iridis regressed in all three eyes in which surgery resulted in complete reattachment of the retina. In one eye with persistent peripheral retinal detachment and in the three remaining eyes that did not undergo revision surgery, rubeosis iridis persisted and was associated with long-term neovascular complications. Final corrected visual acuity was 20/70 to 20/400 in three eyes with total retinal reattachment and no light perception to hand motions in four eyes with persistent peripheral retinal detachment and rubeosis iridis. CONCLUSION: Visually significant neovascular complications may occur in eyes that develop rubeosis iridis associated with peripheral retinal detachment after retinal detachment surgery in nondiabetic patients. Successful repair of the peripheral retinal detachment may induce regression of rubeosis iridis, reduce associated complications, and improve the long-term prognosis of these eyes. PMID- 9744372 TI - Latanoprost treatment for glaucoma: effects of treating for 1 year and of switching from timolol. United States Latanoprost Study Group. AB - PURPOSE: To determine the efficacy and safety of latanoprost treatment for 1 year in glaucoma patients, and to evaluate the effects of switching from timolol to latanoprost therapy. METHODS: Latanoprost 0.005% was topically applied once daily without masking for 6 months in 223 patients with elevated intraocular pressure after previous treatment with latanoprost once daily or 0.5% timolol twice daily for 6 months in a multicenter, randomized, double-masked, parallel group study. RESULTS: Compared with baseline values before treatment, a significant (P < .0001) diurnal reduction in intraocular pressure of 6 to 8 mm Hg was maintained with minimal fluctuation for the duration of treatment. When treatment was switched from timolol to latanoprost, intraocular pressure was reduced by 1.5 +/- 0.3 mm Hg (mean +/- SEM; 8% change in intraocular pressure; 31% of the intraocular pressure reduction produced by timolol; P < .001) compared with the change in intraocular pressure in patients remaining on latanoprost therapy. Of the patients initially enrolled, 95% successfully completed treatment. There was a slight overall increase in conjunctival hyperemia in patients who switched from timolol to latanoprost, but no change in those who continued latanoprost. The timolol-induced reduction of resting heart rate returned to baseline levels after switching to latanoprost. Of the 247 patients treated with latanoprost during the masked and/or open-label studies, 12 (5%) demonstrated a definite (n = 4) or possible (n = 8) increase in iris pigmentation. CONCLUSIONS: Latanoprost is a well-tolerated ocular hypotensive agent that appears to be more effective than timolol in reducing intraocular pressure. The increase in iris pigmentation appears to be harmless but requires further investigation. PMID- 9744373 TI - Clinical efficacy and safety of brinzolamide (Azopt), a new topical carbonic anhydrase inhibitor for primary open-angle glaucoma and ocular hypertension. Brinzolamide Primary Therapy Study Group. AB - PURPOSE: To determine the intraocular pressure-lowering efficacy and safety of brinzolamide 1.0%, compared with dorzolamide 2.0% and timolol 0.5%. METHODS: A multicenter, double-masked, prospective, parallel-group study was conducted to compare brinzolamide 1.0%, administered two and three times a day, dorzolamide 2.0% three times a day, and timolol 0.5% twice a day in 572 patients with primary open-angle glaucoma or ocular hypertension. The primary end point was diurnally corrected intraocular pressure reduction from baseline, evaluated at both peak and trough times during a 3-month period. RESULTS: Mean intraocular pressure changes after twice daily (-3.8 to -5.7 mm Hg) and three times daily (-4.2 to 5.6 mm Hg) dosing with brinzolamide 1.0% were statistically equivalent (confidence limit < or = 1.5 mm Hg) to each other and also to dorzolamide 2.0% three times a day (-4.3 to -5.9 mm Hg). The range of intraocular pressure change with timolol 0.5% twice daily was -5.2 to -6.3 mm Hg. Clinically relevant intraocular pressure changes (reduction > or = 5 mm Hg or intraocular pressure < or = 21 mm Hg) were observed in up to 75.7% of patients taking brinzolamide twice daily and in up to 80.1% taking brinzolamide three times daily. Treatment with brinzolamide 1.0% was safe, comfortable, and well tolerated. The incidence of ocular discomfort (burning and stinging) on instillation of brinzolamide (twice daily, 1.8%; three times daily, 3.0%) was significantly less (P = .000) compared with treatment with dorzolamide (16.4%). CONCLUSIONS: Brinzolamide 1.0% produced clinically relevant intraocular pressure reductions in substantial numbers of patients. Brinzolamide's effectiveness equaled that of dorzolamide 2.0% and it produced less ocular discomfort (burning and stinging) on instillation. PMID- 9744374 TI - Quantitative assessment of macular edema with retinal vein occlusion. AB - PURPOSE: To evaluate the retinal thickness analyzer, a new image analyzer involving laser-slit biomicroscopy, for quantitative assessment of retinal thickness in patients with macular edema secondary to retinal vein occlusion. METHODS: Fifty-eight patients (central retinal vein occlusion, 22 eyes; branch retinal vein occlusion, 36 eyes) with a clinical diagnosis of macular edema associated with retinal vein occlusion were evaluated. Retinal thickness analysis was performed with the retinal thickness analyzer. A green helium-neon laser slit directed into the eye was scanned to generate optical cross-sections of the retina. Images were analyzed by an automated software algorithm, and a detailed map of the retinal thickness was obtained. RESULTS: Laser-slit images obtained with the retinal thickness analyzer in patients with macular edema associated with retinal vein occlusion disclosed intraretinal changes such as retinal edema, cystoid spaces, and hemorrhages. Linear regression analysis showed a significant relationship between the macular thickness in the central fovea and corrected visual acuity reported on a logMAR scale (P = .002, adjusted R2 = 0.40 in central retinal vein occlusion; P = .002, adjusted R2 = 0.34 in branch retinal vein occlusion). CONCLUSION: We confirmed the capability of the retinal thickness analyzer to provide rapid and precise evaluation of macular thickening in patients with retinal vein occlusion. PMID- 9744375 TI - Malattia leventinese: refinement of the genetic locus and phenotypic variability in autosomal dominant macular drusen. AB - PURPOSE: To study the phenotypic variability in patients inheriting the disease gene for malattia leventinese (dominant macular drusen) and refine the localization of the gene. METHODS: A family with dominant radial drusen was ascertained and studied with clinical examination and DNA linkage analysis. Inheritance of the disease gene was determined by DNA analysis and used to document the variability in phenotypic expression. RESULTS: Fifty family members were studied with fundus photography and genotyping. Linkage analysis showed that the disease in this family was linked to chromosome 2p16-21 with a maximum lod score of 3.72 at D2S2153. An affected patient with obligate recombinations allowed refinement of the disease interval to a 6.2-cM region between D2S2227 and D2S378. The phenotype of older affected patients varied from severe geographic atrophy or subretinal fibrosis to a single druse adjacent to the optic disk. Small and medium-sized, nonradial, and soft macular drusen seen in four older individuals in the family were not specifically associated with the disease haplotype. CONCLUSIONS: Refinement of the localization of the gene for malattia leventinese will facilitate its positional cloning. Genotypic documentation of the variable expression of the disease shows that a single, large, subretinal druse adjacent to the optic disk is consistent with inheritance of the disease gene. Soft macular drusen in low abundance were not specifically associated with inheritance of the disease gene. These results will facilitate the genetic counseling of patients with malattia leventinese. It is unknown what proportion of age-related macular degeneration arises from mutations in disease genes for dominant drusen. PMID- 9744376 TI - Quantification of aqueous melanin granules in primary pigment dispersion syndrome. AB - PURPOSE: Aqueous melanin granules are essential in the pathogenesis of pigment dispersion syndrome and pigmentary glaucoma. We quantified aqueous melanin granules with the laser flare-cell meter in patients with pigment dispersion syndrome, assessed the measurement reproducibility, and correlated the numbers with clinical findings. METHODS: Aqueous melanin granules were counted by means of the cell count mode of the laser flare-cell meter (KOWA FC-1000; Kowa, Tokyo, Japan) in 42 eyes of 21 patients with primary pigment dispersion syndrome under three conditions (undilated pupils, dilated pupils, after exercise). The reproducibility of the measurements was determined with the intraclass correlation coefficient. A control group of 40 age- and sex-matched eyes was also examined after pupillary dilation. The results were correlated with biomicroscopic findings in eyes with pigment dispersion syndrome (retrocorneal Krukenberg spindle, iris transillumination, pigmentation of trabecular meshwork). RESULTS: Numerous aqueous melanin granules were detected in eyes with pigment dispersion syndrome (mean, 2.9 +/- 3.7 granules/0.075 mm3) but only small numbers were counted in normal eyes (0.2 +/- 0.3, P < .001). Medical pupil dilation caused an additional increase of aqueous melanin granules in pigment dispersion syndrome (6.3 +/- 5.3, P < .001), but not undilated exercise (climbing stairs) (2.9 +/- 3.7, P > .5). The reproducibility of the measurements was very high (intraclass coefficient >0.92). The number of melanin granules correlated with the degree of Krukenberg spindle (r = .61, P = .004) and with iris transillumination (r = .69, P = .001). CONCLUSIONS: Quantification of aqueous melanin granules yields reproducible results and shows increased numbers in pigment dispersion syndrome, especially after pupillary dilation. Aqueous melanin granule quantification may be useful for evaluating eyes with pigment dispersion syndrome and for assessing treatment effects. PMID- 9744377 TI - Spontaneous remission in patients with essential blepharospasm and Meige syndrome. AB - PURPOSE: To determine the frequency of remission in patients with essential blepharospasm and Meige syndrome. METHOD: Retrospective analysis of patients by means of questionnaire in an institutional setting. RESULTS: Among 238 patients diagnosed in a single institution as having essential blepharospasm or Meige syndrome, 27 (11.3%) claimed to be symptom free without curative surgery. The average time between onset and resolution in the patients who claimed remission was 4.85 years (range, 3 months to 22 years). Among these patients, the duration of remission averaged 6.33 years (range, 1 year to 14 years). CONCLUSION: Patients with essential blepharospasm or Meige syndrome have a small but definite potential for spontaneous remission of symptoms, particularly within the first 5 years after onset of symptoms. PMID- 9744378 TI - Association between MICA gene A4 allele and acute anterior uveitis in white patients with and without HLA-B27. AB - PURPOSE: Acute anterior uveitis is strongly associated with the HLA-B27 antigen and triggered by the involvement of some external factors. However, it is uncertain whether HLA-B27 itself or other gene(s) near the HLA-B region in a linkage disequilibrium with HLA-B27 predispose to this uveitis. We therefore investigated microsatellite polymorphism in the transmembrane region of the major histocompatibility complex class I chain-related gene A (MICA), located 47 kilobases (kb) on the centromeric side of the HLA-B gene on the short arm of chromosome 6 within 6p21.3. METHODS: We examined the following patients for MICA gene polymorphism by means of polymerase chain reaction and subsequent automated fragment detection by fluorescent-based technology: 64 (37 HLA-B27-positive and 27 HLA-B27-negative) whites with acute anterior uveitis, 74 (67 HLA-B27-negative and 7 HLA-B27-positive) ethnically matched random controls, and 36 HLA-B27 positive healthy controls. RESULTS: The microsatellite allele consisting of four repetitions of GCT/AGC (designated A4 allele) was present at the significantly higher phenotype frequency (71.9%) in the patient group than in the ethnically matched random control group (13.5%) (P < .0000001, corrected P < .0000001). The A4 allele was strongly linked to HLA-B27 in a white population. However, the A4 allele was also found at the significantly higher phenotype frequency (37.0%) even in the HLA-B27-negative patient group than in the ethnically matched HLA-B27 negative control group (4.5%) (P = .0086, corrected P = .043). CONCLUSIONS: These results suggest that the MICA gene itself or other nearby gene(s) linked to the MICA A4 allele may be involved in the development of acute anterior uveitis in a white population. PMID- 9744379 TI - Pigment dispersion syndrome. PMID- 9744380 TI - Branch retinal vein obstruction secondary to retinal arteriovenous communication. AB - PURPOSE: To document a branch retinal vein obstruction secondary to a congenital arteriovenous communication. METHOD: Case report of a young patient with retinal arteriovenous communication. RESULTS: A 12-year-old girl had a grade 2 retinal arteriovenous communication in her right eye. She was asymptomatic and was subsequently followed up. Magnetic resonance imaging of the brain was normal and disclosed no signs of Wyburn-Mason syndrome. Nine years later, she developed a branch retinal vein obstruction in the area of the arteriovenous communication. Six months later, the patient was free of secondary complications of branch retinal vein obstruction; however, she is being followed up to detect any retinal or iris neovascularization. CONCLUSION: Awareness of retinal vascular obstruction associated with arteriovenous communication may help its timely recognition, as well as prompt treatment of potential complications, such as retinal and iris neovascularization. PMID- 9744381 TI - Giant pigment epithelial tear and exudative retinal detachment complicating choroidal effusions. AB - PURPOSE: To report a case of a giant pigment epithelial tear and transient exudative retinal detachment occurring in a patient with hypotonic choroidal effusions after trabeculectomy. METHODS: Case report and brief literature review. RESULT: The retinal and choroidal detachments settled, disclosing an extremely large crescentic pigment epithelial defect in the temporal midperiphery. CONCLUSIONS: An exudative retinal detachment secondary to a peripheral pigment epithelial rip may complicate choroidal effusions. Recognition of this association may prevent unnecessary investigation or surgery. PMID- 9744382 TI - The classic form of granular corneal dystrophy associated with R555W mutation in the BIGH3 gene is rare in Japanese patients. AB - PURPOSE: To identify the BIGH3 gene mutation in 10 unrelated Japanese individuals with granular corneal dystrophy. METHODS: Genomic DNA was obtained from each patient's leukocytes. Exons 4 and 12 of the BIGH3 gene were amplified by polymerase chain reaction and were directly sequenced. RESULTS: Nine of these patients were found to have the R124H mutation, whereas only one had the R555W mutation. Slit-lamp examination showed that the granular corneal dystrophy associated with each mutation is different. CONCLUSIONS: These results, together with our previous findings, show that the classic form of granular corneal dystrophy associated with the R555W mutation is rare in Japanese patients, whereas granular corneal dystrophy accompanied by amyloid deposits and associated with the R124H mutation, Avellino corneal dystrophy, is more common. Direct examination may be insufficient in the proper diagnosis of corneal dystrophy, and BIGH3 mutation analysis may be required. PMID- 9744383 TI - POEMS syndrome: an unusual cause of bilateral optic disk swelling. AB - PURPOSE: To alert ophthalmologists to POEMS syndrome as an unusual cause of optic disk swelling. METHOD: Case report. A 25-year-old white woman developed scotomata and optic disk swelling, and systemic signs and symptoms of POEMS syndrome. RESULTS: Bone marrow biopsy disclosed multiple myeloma. Despite treatment with corticosteroids, radiation, and marrow transplantation, the patient died from multi-organ failure. CONCLUSION: Because POEMS syndrome typically affects older male patients, this is an unusual case of multiple myeloma in conjunction with POEMS syndrome. PMID- 9744384 TI - Precipitation of hard exudate after resorption of intraretinal edema after treatment of retinal branch vein occlusion. AB - PURPOSE: To present evidence that precipitation of hard exudate in the retina can occur after reduction of macular edema by successful photocoagulation treatment for retinal branch vein occlusion. METHODS: A 59-year-old woman received argon laser retinal photocoagulation treatment for retinal branch vein occlusion over the primarily involved engorged area of the retina. Fundus changes were documented by fundus photography and optical coherence tomography. RESULTS: Before treatment, an extensive intraretinal edema with cavity formation and a small serous foveal detachment extended outside the engorged area of the retina. Three months after treatment, all retinal thickening had disappeared, but a markedly increased amount of intraretinal hard exudate was seen adjacent to the treated area. CONCLUSIONS: Precipitation of hard exudate can occur after reduction of macular edema by successful retinal photocoagulation treatment, presumably because water and highly soluble electrolytes are resorbed faster than lipids. This phenomenon should be distinguished from accumulation of hard exudate resulting from disease progression. PMID- 9744385 TI - Herpes simplex encephalitis and bilateral acute retinal necrosis syndrome after craniotomy. AB - PURPOSE: Acute retinal necrosis (ARN) syndrome is associated with members of the herpes virus family, but the mechanisms of infection remain unclear. The purpose of this study is to report a unique case of acute retinal necrosis syndrome associated with herpetic encephalitis in order to elucidate possible factors involved in herpetic central nervous system disease. METHOD: Case report. RESULTS: A 64-year-old woman who developed acute herpes simplex virus encephalitis associated with bilateral acute retinal necrosis syndrome after craniotomy for resection of a suprasellar craniopharyngioma is presented. The results of lumbar puncture, magnetic resonance imaging, and ophthalmologic examination are consistent with herpetic infection. The origin of acute retinal necrosis syndrome and the association of acute retinal necrosis syndrome with encephalitis are reviewed. CONCLUSIONS: After craniotomy, we hypothesize reactivation of previously latent herpes simplex virus in the area of the inferior frontal lobe and optic chiasm. Reactivated virus may have migrated to the retina by axonal transport, through the optic nerves, to produce the acute retinal necrosis syndrome. PMID- 9744386 TI - Macular folds and poor vision associated with zone III retinopathy of prematurity. AB - PURPOSE: To describe two cases of zone III retinopathy of prematurity associated with macular folds and poor vision. METHOD: Case reports. RESULTS: Two premature infants with zone III retinopathy of prematurity developed clinically notable elevation of the neovascular ridge with macular folds and poor vision. CONCLUSIONS: Retinopathy of prematurity in zone III may lead to compromised anatomic and functional outcomes. Notable elevation of the neovascular ridge may be an important risk factor for an adverse outcome. PMID- 9744388 TI - A dummy orbit for training in diagnostic procedures and laser surgery with enucleated eyes. AB - PURPOSE: A dummy orbit for enucleated eyes was developed for training residents in diagnostic and surgical procedures. METHODS: Porcine eyes were used to construct the model. The device is made of black anodized metal. The final prototype was tested for ease of use and optical quality. Principal elements are an adjustable eye support, a cylinder, and a removable ring. RESULTS: The dummy orbit and enucleated eye allow direct and indirect ophthalmoscopy; inspection with contact lenses, including gonioscopy; and retinal laser coagulation using contact lenses. Laser trabeculoplasty, argon laser trabeculotomy, Nd:YAG laser iridectomy, and Nd:YAG laser capsulectomy are possible, as well as training in tonometry and ultrasonography techniques. CONCLUSION: The dummy orbit is a valuable aid for teaching and training in diagnostic and laser surgery procedures with nondissected enucleated eyes. PMID- 9744387 TI - Intraocular inflammation and the diffuse infiltrative lymphocytosis syndrome. AB - PURPOSE: To report a case of bilateral panuveitis associated with diffuse infiltrative lymphocytosis syndrome (DILS). METHODS: Case report. A 35-year-old woman with known human immunodeficiency virus (HIV) infection developed bilateral uveitis with retinal periphlebitis in association with diffuse infiltrative lymphocytosis syndrome. RESULTS: Detailed comprehensive examination and investigations were performed and recognized causes of panuveitis excluded. CONCLUSION: To the best of our knowledge, we report the first case of retinal findings associated with diffuse infiltrative lymphocytosis syndrome. PMID- 9744389 TI - Multifocal electroretinogram indicates visual field loss in acute zonal occult outer retinopathy. AB - PURPOSE: To report alterations of electrophysiologic tests, including the multifocal electroretinogram, in a case of acute zonal occult outer retinopathy. METHOD: We recorded photopic, scotopic, and single-flash electroretinograms and a multifocal electroretinogram in a 47-year-old woman with acute zonal occult outer retinopathy in the right eye. RESULTS: Her visual acuity was 20/20 in the right eye throughout the follow-up period. The electroretinograms indicated retinal impairment of the right eye, predominantly in the cones. The multifocal electroretinogram showed reduced responses corresponding to the visual field defect of the static perimetry. CONCLUSIONS: In acute zonal occult outer retinopathy, impairment of the retinal area results in a visual field defect. The multifocal electroretinogram can be useful in determining the location of the defect. PMID- 9744390 TI - Unilateral blindness after ipsilateral prophylactic transcranial optic canal decompression for fibrous dysplasia. AB - PURPOSE: To report a case of unilateral blindness after ipsilateral prophylactic transcranial optic canal decompression for fibrous dysplasia. METHODS: Case report. A 37-year-old woman with fibrous dysplasia underwent a prophylactic left optic canal decompression. Preoperatively, the left eye had a visual acuity of 20/20. RESULTS: Four days after apparently uncomplicated left optic canal decompression, the left eye lost all light perception. Blindness in the left eye persisted despite surgical exploration and transcranial optic nerve sheath fenestration. CONCLUSION: Although prophylactic optic canal decompression warrants consideration in selected patients with fibrous dysplasia, notable risks are associated with this surgery. PMID- 9744391 TI - Indocyanine green angiographic features in posterior scleritis. AB - PURPOSE: To determine choroidal involvement in posterior scleritis by examining indocyanine green angiographic features. METHODS: Indocyanine green angiography was performed according to a standard uveitis angiographic protocol in eight consecutive patients with posterior scleritis. Indocyanine green angiography data were compared to fundus color photographs, red-free photographs, and fluorescein angiography. RESULTS: The principal indocyanine green angiographic feature was diffuse zonal choroidal indocyanine green hyperfluorescence in the intermediate (+/-10 minutes) and late (+/-40 minutes) phases of angiography present in all eight patients who regressed in response to anti-inflammatory therapy. In four patients (two with massive subretinal exudation), additional fluorescing pinpoints were present in the zonal hyperfluorescent areas. Additional features included irregular delayed choroidal perfusion (five of eight patients)- irregularly distributed dark dots that were present up to the intermediate phase and becoming isofluorescent in the late phase that gave a mottled aspect to the choroid--and enlargement of draining choroidal veins. In bilateral patients, clinical features and indocyanine green angiography signs were roughly symmetric. CONCLUSIONS: In posterior scleritis, indocyanine green angiography allowed us to identify areas of choroidal hyperfluorescence, possibly indicating areas of maximal inflammatory activity, and demonstrated regression of hyperfluorescence in response to therapy. Indocyanine green angiography was useful in assessing the extent of choroidal involvement and will probably serve as one of the follow-up parameters for disease evolution and response to therapy. PMID- 9744392 TI - Pupil-sparing third nerve palsy associated with sildenafil citrate (Viagra). AB - PURPOSE: To report unilateral pupil-sparing third nerve palsy after use of sildenafil citrate (Viagra). METHOD: Case report. RESULTS: A 56-year-old man with a history of tobacco abuse was treated for erectile dysfunction. Viagra, 50 mg, was taken once without adverse effect. Three weeks later, the patient took a second dose of Viagra (50 mg); 36 hours later he experienced a complete pupil sparing third nerve palsy. Erythrocyte sedimentation rate, blood glucose level, magnetic resonance imaging, and magnetic resonance angiography were normal. CONCLUSION: In a patient with microvascular disease, use of sildenafil may be associated with pupil-sparing third nerve palsy. PMID- 9744393 TI - An evaluation of neural invasion in esophageal cancer. AB - It is well known that the operative results for esophageal cancer, especially thoracic esophageal cancer, are not favorable. We analyzed the relationship between neural invasion (NI) and histopathologic factors and recurrence types in 104 patients who underwent resection of esophageal cancers with T2 or greater depth of invasion of the esophageal wall. The implications of NI as a prognostic indicator were also examined. Of the 104 patients, 48 (46.2%) were NI-positive (NI(+)) and 56 (53.8%) were NI-negative (NI(-)). The NI(+) patients had a higher ratio of type 3 cancer. Concerning the histopathologic factors, there was a significant relationship between NI and lymph node metastasis (N) and between NI and lymphatic vessel invasion (ly) (P < 0.05). Examining the types of recurrence, namely hematogenous, lymphogenous, and local/stump, as well as pleural or peritoneal dissemination, a relationship was observed between lymphogenous recurrence and N or ly, and between local/stump recurrence and NI. The prognosis of the NI(+) patients was significantly different from that of the NI(-) patients. According to a multivariate analysis, NI and N were significant prognostic factors. These findings demonstrate that NI is an important prognostic factor closely related to local recurrence in patients with esophageal cancer. Thus, when treating advanced esophageal cancer with T2 or greater depth of invasion, NI and lymph node excision should be considered. PMID- 9744394 TI - Relationship between nodal stage and the number of dissected perigastric nodes in gastric cancer. AB - To evaluate the rationality of the current nodal staging system in gastric cancer, we retrospectively analyzed 152 patients with perigastric node involvement localized to a single station, in whom the route of metastasis to distant nodes was limited. No significant differences in pathology or survival were observed between patients with stage n1 and those with stage n2-3 nodal involvement, but the mean (standard deviation) number of perigastric nodes dissected was 22.6 (12.6) in those with stage nl involvement and 18.5 (9.5) in those with stage n2-3 involvement (P = 0.04). When perigastric node involvement was localized to station 3, the mean number of dissected station 3 nodes was 7.7 (4.2) in nl patients and 5.3 (2.8) in n2-3 patients (P = 0.04). This tendency was also observed in patients with perigastric node involvement limited to either station 1 (P = 0.08) or station 6 (P = 0.11). Thus, patients with fewer perigastric nodes may have more lymphatics that bypass perigastric nodes and empty directly into distant nodes, increasing the likelihood of skip metastases. The number of positive nodes, affected to a lesser degree by lymphatic distribution than the location of positive nodes, should be incorporated into the staging criteria. PMID- 9744395 TI - Increased serum concentrations of soluble tumor necrosis factor receptor I in noncachectic and cachectic patients with advanced gastric and colorectal cancer. AB - The serum levels of soluble tumor necrosis factor receptor I (sTNF-RI) were measured in 74 noncachectic patients including 42 with gastric cancer and 32 with colorectal cancer, as well as in 39 patients with severe cachexia and 15 healthy volunteers. The sTNF-RI levels increased with the advance of disease, being highest in the cachectic patients. The levels were inversely correlated with the serum concentrations of nutritional parameters such as prealbumin, transferrin, retinol binding protein, and the percentages of CD3(+) cells in the peripheral blood lymphocytes, and positively correlated with the serum concentration of immunosuppressive acidic protein (IAP) and soluble interleukin-2 receptors. These findings suggest that sTNF-RI could be an important prognostic factor to predict the advance of gastric and colorectal cancers and deterioration of the patient's nutritional and immune activity. PMID- 9744397 TI - Surgery for abdominal aortic aneurysms associated with malignancy. AB - Of 148 patients treated for abdominal aortic aneurysms (AAA), 33 (22%) also had cancer. According to the classification of Szilagyi, there were 13 patients in group I, 19 in group II, and 1 in group IV. In group I, the mean interval between the cancer and AAA operations was 7 years (range 1-14 years). Aneurysmectomy was performed in 9 patients, wrapping in 2, and no operation in 2. In group II, a two stage operation was performed in 8 patients, a single-stage operation in 4, only surgery for cancer in 4, and no operation in 3. Of 4 patients undergoing single stage operations, 3 had colorectal cancer, and there were no postoperative complications such as graft infection or anastomotic breakdown. In group I, 6 of 13 patients died, but there were no cancer deaths. In group II, 9 of 19 patients died, 6 from progressive cancer. The group IV patient also died of cancer. These results suggest that if a patient can tolerate surgery for both diseases, a single-stage operation is preferable. PMID- 9744396 TI - Complications following reparative surgery for aortic coarctation or interrupted aortic arch. AB - Repair of aortic coarctation or interrupted aortic arch continues to be associated with major long-term morbidity. Thus, we conducted a review of 87 consecutive patients who underwent aortic arch repairs, focusing particular attention on the complications that developed. A two-stage strategy was employed if cardiac lesions were associated. The median age at surgery was 1.5 months with a range of 12 h to 56 years. The aortic arch was repaired using end-to-end anastomosis, subclavian flap aortoplasty, subclavian arterial turning-down aortoplasty, patch aortoplasty, tube graft interposition, or other methods. There were 10 patients who died soon after repair, and all of whom had complex cardiac anomalies. Of the remaining 77 patients, 8 developed recurrent stenosis. These 8 patients were all similar in age, being under 3 months old, and weighing 4 kg or less. A multivariable analysis of the infants identified interrupted aortic arch as an independent risk factor for the development of this complication with an odds ratio of 6.45. Complications following prosthesis-free techniques were similar in prevalence and timing. All reinterventions were mortality-free, but catheter dilation and patch aortoplasty were not always successful. Three extraanatomic bypasses were successfully performed, and one adult who had undergone a previous graft and pseudoaneurysm operation was successfully treated with an extraanatomic bypass. These findings led us to conclude that the initial repair should be performed without a prosthesis, and that reintervention for stenosis should combine catheter dilation and extraanatomic bypass. PMID- 9744398 TI - Pulmonary circulatory parameters as indices for the early detection of acute rejection after single lung transplantation. AB - We investigated the relationship between the changes in the pulmonary blood flow and histology during acute rejection following single lung transplantation. In single lung transplantation using adult mongrel dogs, immunosuppression with cyclosporine and azathioprine was discontinued after postoperative day 14 to induce rejection. Doppler flow probes were placed adjacent to the ascending aorta and the left pulmonary artery to measure the blood flow on a daily basis. In addition, chest roentgenograms were also examined daily. The pulmonary pressure was measured using a Swan-Ganz catheter prior to and following the induction of rejection. Open lung biopsies were performed when the left pulmonary artery flow decreased to half of the prerejection value. The pulmonary artery flow decreased to 14.3% of the aortic flow 5 days after the discontinuation of immunosuppression. The graft pulmonary vascular resistance increased significantly compared to the prerejection values (P < 0.001). This was not accompanied by any abnormalities on chest roentgenography. The histology was consistent, with marked perivascular lymphocytic infiltration with little alveolar or interstitial changes. During rejection, the increased pulmonary vascular resistance in the graft was probably the result of perivascular inflammatory cell infiltration, which was seen prior to changes on chest roentgenography. Changes in the left pulmonary artery flow and histology thus appear to be closely correlated in the early stages of acute rejection. PMID- 9744399 TI - Ultrastructural and hormonal changes in the contralateral adrenal gland in unilateral adrenal gland ischemia: an experimental study in rats. AB - While it is well known that unilateral tissue ischemia may result in contralateral damage in some paired organs, there is no universally accepted mechanism to explain why these contralateral changes occur. The aim of the present study was to investigate the ultrastructural and hormonal changes that occur in the contralateral nonischemic adrenal gland after unilateral ischemia of an adrenal gland in a rat model. The animals were divided into four groups of four rats each; namely, a control group which received a sham operation without any ischemic insult, a 2-h ischemic group, a 4-h ischemic group, and an 8-h ischemic group. The left adrenal blood vessels were ligated in all ischemia groups and blood samples were taken for hormonal study 2, 4, and 8 h later, after which bilateral adrenalectomy was performed to determine the ultrastructural changes. The plasma concentrations of aldosterone and cortisol were determined by radioimmunoassays. There was an increase in both aldosterone and cortisol levels related to the duration of the ischemia, but the differences among the groups were not statistically significant. Contralateral ultrastructural damage such as heterochromatin in nuclei, mitochondrial degeneration, endoplasmic reticulum cisternal widening, increased lipid droplets, and lysosomes, were demonstrated electron-microscopically after unilateral adrenal ischemia. PMID- 9744400 TI - Inhibition of tumor growth and microvascular angiogenesis by the potent angiogenesis inhibitor, TNP-470, in rats. AB - The antiangiogenic effects of TNP-470 on the neovascularization of tumors were studied by examining ultrastructural alterations in the vasculature and interstitial fluid pressure (IFP) of tumors. Wistar rats were first inoculated subcutaneously (s.c.) with the Walker 256 carcinosarcoma cell line, then either vehicle medium or TNP-470, 30 mg/kg, was injected s.c. on day 1. A tumor growth assay, the necrotic area, and the IFP in the tumor were all measured on day 12. The antiangiogenic effects of TNP-470 were studied by scanning electron microscopic images of tumor vascular casts. TNP-470 was observed to inhibit tumor growth and increase the necrotic area significantly. In the TNP-470-treated group, the IFP in the superficial layer, defined as 2-3 mm from the tumor capsule, and in the deep layer, defined as 8-10 mm from the tumor capsule, were significantly higher than the corresponding values in the control. Moreover, vascular casts showed a significant reduction in the budding of sprouts in the superficial layer, and a decrease in the maximum diameter of the tumor vessels in the deep layer. It is possible that the higher IFP in the TNP-470-treated tumors might have prevented tumor vessel dilation. The findings of this study demonstrated that TNP-470 inhibited the budding of tumor vessel sprouts, and increased the IFP. These processes seem to act synergistically to suppress tumor angiogenesis. PMID- 9744401 TI - Clinical effect of immunochemotherapy for a patient with advanced gallbladder cancer: report of a case. AB - A 56-year-old woman was admitted presenting with a sensation of abdominal fullness. She was diagnosed to have advanced gallbladder cancer with carcinomatous peritonitis, as well as lymph node and liver metastases. We obtained highly purified tumor cells and tumor-infiltrating lymphocytes (TIL) from extirpated cervical lymph nodes and peritoneal effusion, and the chemosensitivity of these cells was tested with an MTT assay. Intensive chemotherapy with cisplatin (CDDP) and 5-fluorouracil (5-FU) was then performed according to the results of the MTT assay. Thereafter, cytotoxic T-lymphocytes (CTL) were induced in mixed cultures of autologous tumor cells and peripheral blood lymphocytes, and adoptive immunotherapy was performed with TIL and CTL. The malignant ascites and metastatic lesions disappeared after the intraperitoneal administration of CDDP and the transfer of TIL and CTL, and subsequently the patient's quality of life improved. This patient could return to work; however, liver metastasis was later observed, and she died 14 months after the initial diagnosis. Combination therapy with anticancer drugs and activated killer cells was thus found to be effective in a patient with advanced gallbladder cancer. PMID- 9744402 TI - Malignant mesothelioma originating in the hepatic falciform ligament: report of a case. AB - We report herein what to our knowledge is the first documented case of malignant mesothelioma arising from the falciform ligament. A 65-year-old woman was admitted to our department for the evaluation of an epigastric mass. An abdominal computed tomography scan, magnetic resonance imaging, and celiac angiogram showed a tumor originating in the falciform ligament. The tumor was subsequently resected, and pathological examination confirmed a diagnosis of malignant mesothelioma originating in the falciform ligament. A metastatic liver tumor was detected 3 years after this operation and a partial liver resection was performed. She is currently off therapy, and no clinical or radiologic evidence of disease has been found for 1 year since her second operation. PMID- 9744404 TI - Spontaneous perforation of the rectum with possible stercoral etiology: report of a case and review of the literature. AB - Stercoral perforation of the colon or rectum is a rare cause of acute abdomen, with fewer than 70 cases documented in the literature. We report herein the case of a 60-year-old man who presented with anuria and epigastric pain with physical signs of peritonitis. An abdominal X-ray showed bilateral subphrenic free air accumulation, and an emergency laparotomy subsequently revealed perforation of the rectum, suggestive of a stercoral cause, which was treated by simple closure after debridement. Following an uneventful postoperative course, he was discharged from the hospital 3 weeks after his operation and is now doing well without having suffered any further gastrointestinal problems. The clinical features, diagnosis, and treatment of the disease are reviewed following the presentation of this case. Surgeons should be aware of the possibility of this fatal disease, despite its rare incidence. Furthermore, it is important to recognize the condition at an early stage because it has a significantly high mortality if not treated early. Conversely, the surgical outcome is satisfactory provided surgery is performed in due time. PMID- 9744403 TI - Protein-losing enteropathy due to thrombophlebitis of the mesenteric vein: report of a case. AB - We report herein the case of a 70-year-old woman with enteropathy accompanied by protein loss, the cause of which was found to be thrombophlebitis of the mesenteric vein. The patient was admitted to our hospital for investigations to determine the cause of hypoproteinemia. She had suffered an episode of left abdominal pain with high fever and vomiting lasting 10 days, 8 months prior to her admission. She also had a 6-year history of uncontrolled diabetes. The alpha1 antitrypsin clearance was 85.7 ml/day, suggesting protein-losing enteropathy. A scintigraphy with 99m-technetium-human serum albumin disclosed protein leakage into the intestine. X-Rays and computed tomography showed a stenotic and thickened area of small intestine 50 cm in length. Thus, a laparotomy was performed to resect this part of the intestine which was found to have undergone past thrombophlebitic changes. Following the operation, the alpha1-antitrypsin clearance decreased to within the normal range and the patient gained 5 kg in weight. PMID- 9744405 TI - Bowel perforation caused by silicone drains: a report of two cases. AB - We recently encountered two patients who suffered bowel perforation due to pressure necrosis caused by an open silicone drain placed in the abdominal cavity during surgery. These perforations healed spontaneously after removal of the drains from the site of perforation. Silicone drains are frequently placed in the abdominal cavity to prevent the collection of fluid or blood following surgery; however, a risk of this complication must be borne in mind. Our review of the English literature revealed eight cases of bowel perforations occurring due to a drainage system; six to closed suction drains, and two to open drainage tubes. Seven of these eight patients underwent repeat laparotomy for peritonitis, while the remaining one, who had a closed suction drain, was managed conservatively following discontinuation of the vacuum. Our experience and the literature review suggest that conservative management may be possible in patients without any signs of generalized peritonitis, although repeat laparotomy is required for those with generalized peritonitis. In conclusion, drains should be placed carefully in the abdominal cavity and removed early after the drainage has decreased. PMID- 9744407 TI - Hemobilia caused by a giant benign hemangioma of the liver: report of a case. AB - We report herein the extremely unusual case of a 39-year-old woman in whom a giant cavernous hemangioma caused hemobilia. Cavernous hemangioma is the most common benign neoplasm of the liver and rarely causes any clinical symptoms or signs, while hemobilia usually occurs secondary to accidental operative or iatrogenic trauma, vascular disease, inflammatory disorders, gallstones, or tumors of the liver. Although invasive or malignant hepatic tumors often result in a communication between the biliary tract and the blood vessels, only one case of hemobilia caused by a benign cavernous hemangioma has ever been reported, but with no details about the patient. Our patient presented to a local hospital with severe melena as the initial main symptom, where ligation of the right hepatic artery was performed. This failed to relieve her symptoms, and she was subsequently referred to our department where a right hepatectomy was performed. Histopathological examination revealed no malignancy combined with the tumor; however, the hemangioma was exposed to the bile duct in segment VIII, which was presumably the cause of the hemobilia. This patient remains in good health almost 6 years after her operation. To the best of our knowledge this is the first case report of hemobilia caused by a cavernous hemangioma, and is accompanied by a detailed analysis. PMID- 9744406 TI - Hirschsprung's disease in an adult patient with familial occurrence: report of a case. AB - Hirschsprung's disease is almost always associated with newborns or infants; however, we report herein the unusual case of a 46-year-old woman in whom the symptoms of Hirschsprung's disease emerged late in adult life. The involved rectosigmoid region was successfully removed by performing Duhamel's operation with a diverting colostomy. After the colostomy was closed, she regained normal defecatory function. She had one male child affected by Hirschsprung's disease of the total colon type who was operated on as a 12-month-old baby. The genetic predisposition of Hirschsprung's disease has been reported, but its mode of inheritance has not yet been clarified. Moreover, most papers on the familial occurrence of this disease have reported that siblings were affected. Our patient was unique for the definite occurrence of the disease in successive generations. The features of Hirschsprung's disease in adults and the familial occurrence are discussed with a review of the literature. PMID- 9744408 TI - Metastatic malignant meningioma of the liver with hypoglycemia: report of a case. AB - We herein present the case of a 68-year-old male who suffered an episode of hypoglycemic shock 2 years after undergoing total removal of a bifrontal parasagittal malignant meningioma. Imaging studies revealed three giant hypervascular tumors with a cystic portion in the right lobe, but no confirmed preoperative diagnosis could be made. At laparotomy, liver tumors were found in the medial segment of the left lobe as well as in the right lobe, and thus an extended right lobectomy was performed. All the resected tumors were histologically diagnosed as metastatic malignant meningiomas of the liver. Despite subsequent transarterial chemoembolization for a recurrence in the residual liver, the patient died 11 months after surgery. To the best of our knowledge, only one other case of a hepatectomy for liver metastases from an intracranial malignant meningioma has been reported in the literature, but there has never been any report of surgical treatment for a metastatic meningeal tumor in the liver associated with hypoglycemia. Although our surgical treatment provided effective palliation, the prognostic significance of a surgical strategy for such patients has yet to be established. PMID- 9744409 TI - Successful laparoscopic cholecystectomy in a child with upper transverse scarring: report of a case. AB - Laparoscopic cholecystectomy (LC) was safely performed for cholelithiasis in a 4 year-old boy who had a long transverse operative scar in the upper abdomen as a result of intestinal surgery performed during the neonatal period. The adhesions beneath the scar were sharply divided and sometimes coagulated, and additional working ports were subsequently placed as the adhesiolysis proceeded. LC was performed in the usual fashion using 5-mm titanium clips, and his postoperative course was uneventful. This case report serves to demonstrate that laparoscopic surgery is feasible even for pediatric patients who have undergone previous major intraabdominal surgery. PMID- 9744410 TI - Successful graft replacement of the descending aorta after an extended reconstruction of the ascending and transverse aorta in a patient with Marfan's syndrome. AB - A 27-year-old man with Marfan's syndrome underwent a total aortic graft replacement in three separate stages. Initially the abdominal aorta was replaced, followed by the ascending aorta and aortic arch, and finally the residual portion. The extensive reconstruction of both the ascending and transverse aorta at the second operation, even though no dissection was present in the aortic arch, reduced the risk of the subsequent operation since the same surgical approach did not have to be used. PMID- 9744411 TI - Venous aneurysm of the cephalic vein: report of a case. AB - We herein report the rare case of a 41-year-old Japanese woman in whom a venous aneurysm in the left cephalic vein was excised under local anesthesia. Histological examination revealed significant diminution in the number and size of muscle and elastic fibers in the aneurysm wall. Conceivably, a combination of endophlebohypertrophy and a congenital focal defect of the elastic and muscle fibers might have contributed to the development of this venous aneurysm. PMID- 9744412 TI - Utilization of the lateral circumflex femoral artery as a midway outflow for aorto-popliteal grafting: report of a case. AB - We describe herein the case of a patient who presented with total occlusion of all the major arteries in the unilateral iliofemoral region, including the distal deep femoral artery, on whom an aortolateral circumflex femoral-popliteal artery sequential bypass was successfully performed. This case report serves to demonstrate that the lateral circumflex femoral artery can provide a suitable midway outflow for aortopopliteal bypass in patients with extensive thrombosis of the iliofemoral arteries. PMID- 9744413 TI - Right coronary artery dissection and acute infarction due to blunt trauma: report of a case. AB - Coronary artery dissection occurring after a nonpenetrating chest trauma is extremely rare. We describe herein the case of a 43-year-old man who suffered traumatic myocardial infarction after an intimal tear of the right coronary artery had been inflicted by a horse stepping on his back. PMID- 9744414 TI - Renin-producing adrenal tumor: report of a case. AB - A 19-year-old man with an adrenal tumor associated with hypertension is herein described. The plasma renin activity (PRA) was markedly elevated and the plasma aldosterone level was also increased. The catecholamine level was within the normal range and the glucagone-regitine test was negative. An angiogram revealed the left renal artery to have no stenotic segments, but instead was curved caudally. Magnetic resonance imaging showed a tumor measuring about 8 cm in diameter to be clearly recognized from the left kidney. The blood pressure did not drop when a calcium-channel blocker was used, but was gradually stabilized with the angiotensin converting enzyme (ACE) antagonist. The adrenal tumor was then removed surgically. Normal adrenal tissue was scarcely recognized to the left of the tumor. After surgery, the PRA decreased and blood pressure stabilized rapidly without the ACE antagonist. The possibility of massive renal renin secretion due to suppression by the adrenal tumor was excluded by a method using an antibody to purified human renin. PMID- 9744415 TI - Multidrug-resistant recurrent breast cancer which responded to medroxyprogesterone acetate showing a remarkable improvement in the quality of life: report of a case and the role of team medical care. AB - We herein report the case of a patient with recurrent breast cancer who showed a remarkable improvement in her quality of life (QOL) as a result of a good response to medroxyprogesterone acetate (MPA). A 43-year-old Japanese woman developed bone metastases 3 years after surgery. Subsequent radiotherapy and chemoendocrine therapy with CAF (cyclophosphamide, adriamycin, 5-fluorouracil) and tamoxifen all failed, and she could not sit up because of bone metastases. The performance status (PS) on admission was grade 4. After admission, delirium accompanied with sensory and visual hallucination caused by intense anxiety occurred, and a continuous consultation by psychiatrists was necessary. MPA treatment at the dose of 1200 mg/day alleviated the bone pain, thus improving her PS to grade 1. Her appetite also improved, while her mental state stabilized. A bone scintigram revealed an improvement of bone metastases, and the tumor markers also returned to normal values. The patient thus showed a pronounced improvement in her QOL due to both MPA treatment and team medical care. The role of the medical staff as well as the importance of their cooperation in achieving an improvement in the QOL of cancer patients is also discussed. PMID- 9744416 TI - The use of transbronchial lung biopsy to establish a diagnosis of benign clear cell tumor of the lung: report of a case. AB - Benign clear cell tumor (BCCT) is an extremely rare pulmonary neoplasm. We report herein a case of BCCT of the lung that was diagnosed by a transbronchial lung biopsy (TBLB). The findings of hematoxylin-eosin and immunohistochemistry obtained by the TBLB were consistent with a diagnosis of BCCT of the lung. Wedge resection was subsequently performed by video-assisted thoracotomy. To the best of our knowledge, this is the first reported case of BCCT diagnosed by TBLB. PMID- 9744417 TI - Surgical management for a malignancy of the digestive organs accompanied with an abdominal aortic aneurysm. AB - Surgical management of patients with simultaneous coexisting malignancy of the digestive organs and an abdominal aortic aneurysm (AAA) remains controversial. In the five patients who underwent the aneurysmectomy first, no complications developed after an aneurysmectomy and a resection of malignancy could be performed within 4 weeks, whereas postoperative complications after the resection of malignancy developed in two of them. Two patients underwent a one-stage operation, in which one was unable to tolerate the two procedures, and no postoperative complications were seen; however, one patient with cardiac dysfunction who first underwent an aneurysmectomy died 3 months after operation due to cardiac and renal failure. These results indicate that the aneurysmectomy first is preferred, when such patients do not have absolute indications of malignancy or AAA; however, a one-stage operation should be chosen when the patients show a disturbance of key organs. PMID- 9744418 TI - Hippocampal PET activations of memory encoding and retrieval: the HIPER model. AB - A meta-analysis of experimentally induced changes in blood flow ("activations") in positron emission tomography (PET) studies of memory has revealed an orderly functional anatomic pattern: Activations in the hippocampal region associated with episodic memory encoding are located primarily in the rostral portions of the region, whereas activations associated with episodic memory retrieval are located primarily in the caudal portions. These findings are based on an analysis of a sample of 54 "hippocampal encoding and retrieval" activations that were culled from an overall database consisting of 52 published PET studies of memory. We refer to this general pattern of rostrocaudal gradient of encoding and retrieval PET activations as the HIPER (Hippocampal Encoding/Retrieval) model. The model suggests a division of memory-related labor between the rostral and caudal portions of the hippocampal formation. Because functional anatomic pattern of encoding and retrieval activation that defines the HIPER model was unprecedented and unexpected, it is difficult to relate the model to what is already known or thought about functional neuroanatomy of episodic memory in the hippocampal regions. The model is interesting primarily because its exploration may yield fresh insights into the neural basis of human memory. PMID- 9744419 TI - Selective hippocampal lesions yield nonspatial memory impairments in rhesus monkeys. AB - Monkeys with removals of medial temporal lobe (MTL) structures are widely recognized as valid models of human global anterograde amnesia, a syndrome that arises consequent to damage to a finite set of brain structures situated in the medial temporal lobe and/or medial diencephalon. However, a comparison of memory deficits in human and nonhuman primates with MTL damage has presented a long standing puzzle. Whereas amnesic patients are impaired in learning object discrimination problems, monkeys with MTL damage are typically not. One possible explanation for this difference is that object discrimination tasks for humans and monkeys differ in that the former but not the latter requires the use of contextual information. If this analysis is correct, monkeys with MTL damage might be disadvantaged in learning to discriminate similar objects presented in different contexts. To test this possibility, we evaluated the effects of excitotoxic lesions of one of the MTL structures, the hippocampus, on the rate of learning of discrimination problems embedded within unique contexts. Monkeys with hippocampal lesions were impaired relative to controls in learning object discrimination problems of this type. These findings strongly support the idea that the difference in the effect on object memory of MTL damage in human and nonhuman primates is due to a difference in the opportunity to employ contextual cues rather than to a difference in the organization of memory. PMID- 9744420 TI - The human perirhinal cortex and recognition memory. AB - The importance of the perirhinal cortex for visual recognition memory performance is undisputed. However, it has not been clear whether its contribution to performance is mainly perceptual, or mainly mnemonic, or whether the perirhinal cortex contributes to both perception and memory. We determined the effects of medial temporal lobe damage that includes complete damage to the perirhinal cortex in two amnesic patients by assessing recognition memory for complex visual stimuli across delays from 0 to 40 s. These patients, as well as six other amnesic patients with damage limited to the hippocampal formation or diencephalic structures, exhibited intact recognition memory at delays of 0-2 s and a delay dependent memory impairment at delays of 6 s and longer. Additionally, the patients with damage to the perirhinal cortex performed worse than the other amnesic patients at delays of 25 s and longer. The findings suggest that the perirhinal cortex is not important for visual perception or immediate memory. In this respect, the findings for perirhinal cortex resemble the findings for other medial temporal lobe structures, including the hippocampus. PMID- 9744421 TI - Superior colliculus and active navigation: role of visual and non-visual cues in controlling cellular representations of space. AB - To begin investigation of the contribution of the superior colliculus to unrestrained navigation, the nature of behavioral representation by individual neurons was identified as rats performed a spatial memory task. Similar to what has been observed for hippocampus, many superior collicular cells showed elevated firing as animals traversed particular locations on the maze, and also during directional movement. However, when compared to hippocampal place fields, superior collicular location fields were found to be more broad and did not exhibit mnemonic properties. Organism-centered spatial coding was illustrated by other neurons that discharged preferentially during right or left turns made by the animal on the maze, or after lateralized sensory presentation of somatosensory, visual, or auditory stimuli. Nonspatial movement-related neurons increased or decreased firing when animals engaged in specific behaviors on the maze regardless of location or direction of movement. Manipulations of the visual environment showed that many, but not all, spatial cells were dependent on visual information. The majority of movement-related cells, however, did not require visual information to establish or maintain the correlates. Several superior collicular cells fired in response to multiple maze behaviors; in some of these cases a dissociation of visual sensitivity to one component of the behavioral correlate, but not the other, could be achieved for a single cell. This suggests that multiple modalities influence the activity of single neurons in superior colliculus of behaving rats. Similarly, several sensory-related cells showed dramatic increases in firing rate during the presentation of multisensory stimuli compared to the unimodal stimuli. These data reveal for the first time how previous findings of sensory/motor representation by the superior colliculus of restrained/anesthetized animals might be manifested in freely behaving rats performing a navigational task. Furthermore, the findings of both visually dependent and visually independent spatial coding suggest that superior colliculus may be involved in sending visual information for establishing spatial representations in efferent structures and for directing spatially-guided movements. PMID- 9744422 TI - NMDA receptor-independent LTP in basal versus apical dendrites of CA1 pyramidal cells in rat hippocampal slice. AB - The ability of hippocampal CA1 basal synapses to express N-methyl-D-aspartate (NMDA) receptor-independent long-term potentiation (non-NMDA LTP) was studied and compared to the simultaneously induced apical dendritic non-NMDA LTP. Non-NMDA LTP in basal and apical dendrites was induced using stimulation pattern similar to the sharp wave-associated CA3 bursts. Basal dendritic non-NMDA LTP was input specific and displayed similar development and magnitude to the apical dendritic non-NMDA LTP. Both apical and basal dendritic non-NMDA potentiations were inhibited by the voltage-dependent calcium channel (VDCC) inhibitor verapamil and the tyrosine kinase inhibitors genistein and levandustin A. However, the difference in the degree and time course of these inhibitions suggests involvement of distinct mechanisms in the two dendritic subfields. PMID- 9744423 TI - Electrophysiological properties of rat hippocampal principal cells are unaltered by prenatal protein malnutrition. AB - There is growing evidence that prenatal protein malnutrition alters the development of the hippocampal formation in rats (Morgane et al., 1993; Galler et al., 1996; Almeida et al., 1996, for reviews). Little is known, however, of the possible functional consequences of prenatal malnutrition on the physiology of principal cells in the hippocampus. We have addressed this issue by comparing the electrophysiological properties of hippocampal neurons (dentate granule cells and CA1 pyramidal cells) in slices prepared from control and from prenatally protein malnourished adult male and female Sprague-Dawley rats. We found no significant effect of the prenatal protein malnutrition insult upon a number of intrinsic membrane properties measured with whole-cell current clamp recordings, including: resting membrane potential, input resistance, and membrane time constant, or on action potential characteristics such as threshold, amplitude, and/or firing frequency. Additionally, we saw no effect of prenatal malnutrition upon extracellular measures of glutamatergic synaptic transmission such as the presynaptic fiber volley, excitatory postsynaptic potential or population spike amplitude at the perforant path to dentate granule cell synapse or at the Schaffer collateral to CA1 pyramidal cell synapse. In conclusion, we have demonstrated that prenatal protein malnutrition does not result in significant alterations of the cellular physiological properties of these two types of principal neurons in the adult rat hippocampus. PMID- 9744424 TI - Glutamate antagonism during secondary deafferentation enhances cognition and axo dendritic integrity after traumatic brain injury. AB - The combination of central fluid percussion traumatic brain injury (TBI) followed 24 h later by a bilateral entorhinal cortical deafferentation (BEC) produces profound cognitive morbidity. We recently showed that MK-801 given prior to TBI in this insult improved spatial memory for up to 15 days. In the present study we examine whether MK-801 treatment of the BEC component in the combined insult model affects cognitive recovery. Two strategies for drug treatment were tested. Fifteen minutes prior to the BEC lesion in the combined insult, rats were given i.p. doses of either 3 mg/kg (acute group) or 1 mg/kg (chronic group) MK-801. The acute group received no further injections, whereas the chronic group received 1 mg/kg MK-801 i.p. twice a day for 2 days post-BEC lesion. Two additional groups of animals received BEC lesion alone and either acute or chronic MK-801 treatment identical with the combined insult cases. Each group was then assessed for spatial memory deficits with the Morris water maze at days 11-15 and 60-64 postinjury. Both acute and chronic MK-801 treatment in the combined insult group significantly reduced spatial memory deficits at 15 days postinjury relative to untreated injured cases (P < .01). This reduction appeared more robust at 15 days and persisted for up to 64 days in the chronically treated group (P < .05). By contrast, neither acute nor chronic MK-801 treatment affected memory performance with the BEC insult alone. Immunocytochemical localization of parvalbumin showed that chronic administration of MK-801 in the combined insult cases attenuated the injury-induced dendritic atrophy of inhibitory neurons in the dentate gyrus and area CA1. Synaptophysin immunobinding revealed that chronic MK-801 treatment of the BEC component of the combined insult normalized the distribution of presynaptic terminals within the dentate gyrus. These results suggest that cognitive deficits produced by head trauma involving both neuroexcitation and deafferentation can be attenuated with chronic application of glutamatergic antagonists during the period of deafferentation injury and that this attenuation is correlated with axo-dendritic integrity. PMID- 9744425 TI - Age-related deficits on the radial maze and in fear conditioning: hippocampal processing and consolidation. AB - Young adult, middle-aged, and old male F-344 rats were assessed for their hippocampal ability. This was accomplished by examining the animals on two different paradigms, each incorporating a simultaneous measure of hippocampal dependent and -independent processing. The animals were fear conditioned and then tested for retention of the conditioning context and tone. This was followed by an 8-arm radial maze task which combined spatial working and cued reference memory elements. The two paradigms are compared in terms of task demands, potential confounds, and validity for aging studies. The results indicate that the performance of the animals on the two tasks is correlated. Age-related deficits limited to the hippocampal aspects of the above tasks were found, with no deficits found in the analogous but hippocampus-independent aspects of these tasks. The function of the hippocampus in incorporating new memories is time related. Therefore, the possibility of age-related changes in consolidation was examined. It has previously been shown on the fear conditioning paradigm that the hippocampus is involved in retention of the aversive context for approximately 28 days. In the present study, an attempt was made to test the animals for retention of the conditioning context both early into the period of consolidation (10 days) and after consolidation should have been completed (52 days). The results indicate that, initially, the old animals show comparable retention to young rats. When examined later, young animals showed a stronger retention of the conditioning context than they had previously. The aged rats, however, did not seem to benefit from this additional period of time and in fact showed a decrease in retention of the conditioning context. The data are interpreted in terms of consolidation, alternative explanations of the data are presented, and suggestions are given for future research. Finally, the implications of such age related changes in hippocampal consolidation on learning and memory are discussed. PMID- 9744426 TI - Neural circuit analysis of spatial working memory: role of pre- and parasubiculum, medial and lateral entorhinal cortex. AB - Using a continuous recognition memory procedure for spatial location information, rats were given sequential presentation of individual arms on a 12-arm maze. Each arm contained a Froot Loop reinforcement the first time it was presented, and latency to traverse the arm was measured. A subset of the arms were repeated, but did not contain reinforcement. Repeated arms were presented with lags ranging from zero to six (from zero to six different arm presentations occurred between the first and repeated presentation). After completion of acquisition training (significantly longer latencies for repeated arms in comparison with the first presentation of an arm), rats received lesions of the medial or lateral entorhinal cortex, pre- and parasubiculum, or served as sham-operated controls. Based on continued postsurgery training and additional tests, the results indicated that rats with pre- and parasubiculum or pre- and parasubiculum plus medial entorhinal cortex produced sustained impairment in performing the task. Medial or lateral entorhinal cortex and control lesions did not display any sustained deficits. The data suggest that working memory for spatial location information is mediated primarily by the pre- and parasubiculum, but not medial entorhinal and lateral entorhinal cortex. PMID- 9744427 TI - Is there a direct projection from perirhinal cortex to the hippocampus? PMID- 9744428 TI - The ICD in France: money problem or donkey syndrome? PMID- 9744429 TI - The use of the block cycle length as a safe and efficient means of interrupting sustained ventricular tachycardia and its pharmacological modification. AB - In nine patients who had inducible monomorphic sustained ventricular tachycardia (VT), rapid pacing was performed in 11 episodes of morphologically distinct VT at progressively shorter cycle lengths and VT was interrupted at a critical cycle length. The VT interrupting critical cycle length was defined as the block cycle length (BCL) and the effect of Class I antiarrhythmic drugs were examined. Both the VT cycle length (VTCL) and the BCL were prolonged after administration of either drug. The overall mean ratio of the BCL to the VTCL was unchanged after procainamide administration, but increased after the use of mexiletine. The ratio, however, varied in individual VTs and the BCL after treatment with Class I antiarrhythmic drugs could not be predicted from the ratio baseline value, although the ratio was always > 60% and the hazard of VT acceleration might be avoided if the BCL is used. PMID- 9744430 TI - Radiofrequency catheter ablation of concealed atrioventricular accessory pathways using a "simultaneous pacing method". AB - The retrograde atrial potential at a successful ablation site is usually obscured by the wide and large ventricular potential during atrioventricular reentrant tachycardia or ventricular pacing, which makes it difficult to determine the appropriate ablation site for a concealed accessory pathway. A pacing maneuver named the "simultaneous pacing method" is proposed herein to differentiate the retrograde atrial potential from the ventricular potential for a successful ablation of the concealed accessory pathway. Catheter ablation was performed in 12 patients with a single left free-wall concealed accessory pathway. The atrial insertion site was determined by the simultaneous pacing method in six patients (group I) and by ventricular pacing in six patients (group II). In the simultaneous pacing method, electrograms recorded during ventricular pacing in the earliest retrograde atrial activation site are a fusion of the ventricular potential and the following retrograde atrial potential. When atrial and ventricular pacings are performed simultaneously (simultaneous pacing), the end portion of the electrograms recorded at the same site is solely the ventricular component, because atrial is activated earlier. The atrial potential can be confirmed during ventricular pacing in comparison with the electrograms during the "simultaneous pacing." Radiofrequency catheter ablation was successful in eliminating conduction through the accessory pathway in all 12 patients. The radiofrequency applications in group I were significantly fewer than those in group II (1.7 +/- 1.0 in group I, 5.3 +/- 3.2 in group II, P < 0.05). The total procedure time in group I was significantly shorter than in group II (57.8 +/- 15.7 vs 106.7 +/- 41.6 mins in group II, respectively, P < 0.05). The fluoroscopy time in group I was significantly shorter than in group II (54.0 +/- 7.9 vs 81.3 +/- 26.3 mins, respectively, P < 0.05). We were able to determine the atrial insertion site of accessory pathways by the simultaneous pacing method. The simultaneous pacing method was useful in eliminating concealed left free-wall accessory pathways. PMID- 9744431 TI - Effects of sensor selection on exercise stroke volume in pacemaker dependent patients. AB - The effects of sensor selection and sensor blending on the cardiovascular response to graded exercise was evaluated in 10 patients (age 74 +/- 2 yrs; 7 men and 3 women) implanted with a dual sensor rate adaptive VVIR pacemaker (Vitatron Topaz model 515). Patients underwent three graded exercise tests (GXT) with sensor programming randomly assigned. For a given graded exercise text the pacemaker was programmed into activity sensing (ACT), QT sensing, or dual sensing (ACT = QT). Data were recorded at rest and during each stage of the graded exercise text. Oxygen uptake (VO2) was measured continuously using a Q Plex I system. Heart rate (HR), stroke volume (SV), and cardiac output (Qc) were measured by impedance cardiography. Systolic time intervals were calculated from simultaneous recordings of the ECG, phonocardiogram, and the impedance cardiogram. In response to the GXT no differences in peak VO2 were observed across the three sensor settings. Regardless of the sensor setting Qc increased linearly with each increment in VO2. The HR response to ACT only pacing was significantly higher than in the other two pacing conditions. During ACT only pacing SV failed to rise in response to exercise. The increased exercise Qc during QT and ACT = QT pacing were mediated by significant increases in both HR and SV. The QT and dual pacing conditions were also associated with longer diastolic filling times. The data indicate that the mechanisms responsible for the increase Qc during exercise were different for ACT versus ACT = QT or QT sensor-driven pacing. PMID- 9744432 TI - Cellular phone interference testing of implantable cardiac defibrillators in vitro. AB - An in vitro study was undertaken to investigate the potential for cellular telephones to interfere with representative models of presently used ICDs. Digital cellular phones (DCPs) generate strong, amplitude modulated fields with pulse repetition rates near the physiological range sensed by the ICD as an arrhythmia. DCPs with Time Division Multiple Access (TDMA) pulsed amplitude modulation caused the most pronounced effect--high voltage firing or inhibition of pacing output of the ICDs. This electromagnetic interference (EMI) occurred only when the phones were within 2.3-5.8 cm of the ICD pulse generator that was submerged 0.5 cm in 0.18% saline. ICD performance always reverted to baseline when the cellular phones were removed from the immediate proximity of the ICD. Three models of ICDs were subjected to EMI susceptibility testing using two types of digital phones and one analog cellular phone, each operating at their respective maximum output power. EMI was observed in varying degrees from all DCPs. Inhibition of pacer output occurred in one ICD, and high voltage firing occurred in the two other ICDs, when a TDMA-11 Hz DCP was placed within 2.3 cm of the ICD. For the ICD that was most sensitive to delivering unintended therapy, inhibition followed by firing occurred at distances up to 5.8 cm. When a TDMA-50 Hz phone was placed at the minimum test distance of 2.3 cm, inhibition followed by firing was observed in one of the ICDs. EMI occurred most frequently when the lower portion of the monopole antenna of the cellular phone was placed over the ICD header. PMID- 9744434 TI - Nonfluoroscopic guidance for catheter placement into the coronary sinus under direct vision using a balloon-tipped cardioscope. AB - The right atrial posterior septum, including the coronary sinus (CS) ostium, is an important landmark in radiofrequency catheter ablation therapy for supraventricular tachycardia or atrial flutter. The anatomical findings around the CS ostium would be useful to determine a target site or line during catheter ablation. The aim of the study was to test the ability of the imaging catheter to identify structures in the posterior septal area of the right atrium and to evaluate the feasibility of guidance for catheter placement in the CS using a cardioscope that we recently developed. In 12 anesthetized dogs, the cardioscope, consisting of a deflectable 7 Fr fiberoptic endoscope with an inflatable and transparent balloon, was introduced into the right atrium via the femoral vein. The cardioscope was manipulated to observe the right atrial posterior septum. A deflectable electrode catheter was inserted via the jugular vein and positioned in the CS under cardioscopic guidance. In 10 of 12 dogs, the right atrial posterior septum, including the CS ostium, and the tendon of Todaro could be anatomically identified by cardioscopy. It was possible to position an electrode catheter in the CS in all 10 dogs under direct vision without fluoroscopy. But the CS ostium could not be detected in the remaining two dogs, although the cardioscope was placed at as many sites as possible. No complication occurred. The balloon-tipped cardioscope appears to be useful in observing the right atrialposterior septum and in guiding an electrode catheter into the CS. PMID- 9744433 TI - Quantification of the modifications in the dominant frequency of ventricular fibrillation under conditions of ischemia and reperfusion: an experimental study. AB - The characteristics of ventricular fibrillatory signals vary as a function of the time elapsed from the onset of arrhythmia and the maneuvers used to maintain coronary perfusion. The dominant frequency (FrD) of the power spectrum of ventricular fibrillation (VF) is known to decrease after interrupting coronary perfusion, though the corresponding recovery process upon reestablishing coronary flow has not been quantified to date. With the aim of investigating the recovery of the FrD during reperfusion after a brief ischemic period, 11 isolated and perfused rabbit heart preparations were used to analyze the signals obtained with three unipolar epicardial electrodes (E1-E3) and a bipolar electrode immersed in the thermostatized organ bath (E4), following the electrical induction of VF. Recordings were made under conditions of maintained coronary perfusion (5 min), upon interrupting perfusion (15 min), and after reperfusion (5 min). FrD was determined using Welch's method. The variations in FrD were quantified during both ischemia and reperfusion, based on an exponential model deltaFrD = A exp ( t/C). During ischemia deltaFrD is the difference between FrD and the minimum value, while t is the time elapsed from the interruption of coronary perfusion. During reperfusion deltaFrD is the difference between the maximum value and FrD, while t is the time elapsed from the restoration of perfusion. A is one of the constants of the model, and C is the time constant. FrD exhibited respective initial values of 16.20 +/- 1.67, 16.03 +/- 1.38, and 16.03 +/- 1.80 Hz in the epicardial leads, and 15.09 +/- 1.07 Hz in the bipolar lead within the bath. No significant variations were observed during maintained coronary perfusion. The fit of the FrD variations to the model during ischemia and reperfusion proved significant in nine experiments. The mean time constants C obtained on fitting to the model during ischemia were as follows: E1 = 294.4 +/- 75.6, E2 = 225.7 +/- 48.5, E3 = 327.4 +/- 79.7, and E4 = 298.7 +/- 43.9 seconds. The mean values of C obtained during reperfusion, and the significance of the differences with respect to the ischemic period were: E1 = 57.5 +/- 8.4 (P < 0.01), E2 = 64.5 +/- 11.2 (P < 0.01), E3 = 80.7 +/- 13.3 (P < 0.01), and E4 = 74.9 +/- 13.6 (P < 0.0001). The time course variations of the FrD of the VF power spectrum fit an exponential model during ischemia and reperfusion. The time constants of the model during reperfusion after a brief ischemic period are significantly shorter than those obtained during ischemia. PMID- 9744435 TI - Outpatient pacemaker procedures in orally anticoagulated patients. AB - Reexamination of surgical practices in the present era of cost containment has led to increased outpatient procedures including pacemaker surgery. While the safety and economic benefits of outpatient pacemaker surgery in nonanticoagulated patients is well documented, results of pacemaker operations in patients maintained on coumadin for thromboembolic prophylaxis have not been evaluated. In patients where complications with pacemaker surgery appeared successive, we have established a low incidence of complications. Recently, we extended this approach to the outpatient setting; this report retrospectively reviews our 4-year experience. During the study period, 150 patients underwent outpatient pacemaker procedures, including 37 patients receiving oral warfarin. There was no difference in the incidence of wound related and wound unrelated complications between patients receiving warfarin and the nonanticoagulated cohort. In addition, no wound hematomas, blood transfusions, or clinically significant bleeding episodes were noted among warfarin recipients. We conclude that pacemaker surgery in patients receiving oral anticoagulation is safe and feasible. The use of the cephalic cutdown technique avoiding blind subclavian punctures, meticulous attention to pocket hemostasis, and the use of small caliber unipolar positive fixation leads appears warranted in this selected group of patients at high risk for perioperative bleeding. PMID- 9744437 TI - Fluoroscopic determination of latissimus dorsi muscle shortening fraction after dynamic cardiomyoplasty. AB - Optimization of the skeletal muscle contraction during cardiac assist is important to achieve maximal cardiac assist and yet avoid overstimulation that may injure skeletal muscle. Dynamic cardiomyoplasty suffers from lack of an objective, reproducible, and accurate technique to measure skeletal muscle shortening fraction after wrap and training of the muscle. A recruitment curve is considered the best way to select the proper stimulation level to achieve supramaximal contraction without overstimulating the muscle. A fluoroscopic technique of determining latissimus dorsi recruitment curve was evaluated in five goats undergoing dynamic cardiomyoplasty with an anterior cardio-subcutaneous wrap. Two pairs of stainless steel targets (0.5 and 1 cm of diameter) were implanted on each side of the muscle wrap. One pair of sonomicrometer crystals was also implanted. Displacement of the targets was measured under fluoroscopy at five different stimulation levels. Correlation coefficients between targets on the inside surface of the wrap and the sonomicrometer crystals, and targets on the outside surface of the wrap and the sonomicrometer crystals were 0.71 (P < 0.05) and 0.60 (P < 0.05), respectively. Targets on the inside surface of the wrap were more accurate than targets on the outside surface of the wrap for measurement of skeletal muscle shortening fraction and establishment of a recruitment curve. Adverse effects from the targets were not observed. PMID- 9744436 TI - Mid- and long-term similarity of ventricular response to paroxysmal atrial fibrillation: digoxin versus placebo. AB - The effects of digoxin on ventricular response during atrial fibrillation (AF) and consequent effects on arrhythmic symptoms have still not been fully explained. This study investigated whether the treatment by digoxin contributes to mid- and long-term stabilization of ventricular cycles in patients with paroxysmal AF. A population of 45 patients with paroxysmal AF underwent 24-hour ECG recordings during each arm of a randomized crossover trial comparing digoxin and placebo. This yielded 30 Holter recordings from 22 patients that contained AF episodes lasting in excess of 2 minutes and with acceptably low Holter noise. Each AF episode was divided into nonoverlapping segments of 30 seconds and the distribution of RR intervals in each segment was compared with the distribution of all other AF segments in the same recording using the Kolmogorov-Smirnov test. The percentage of tests that revealed significant differences at levels of P < or = 0.01, and P < or = 0.001 were sorted according to the time between the segments compared. The comparisons of these results were performed between: (a) all recordings on placebo (n = 16) and all recordings on digoxin (n = 14), and (b) between recordings on placebo and on digoxin in 8 patients in whom paired analysis was possible. Adjacent AF segments (distance 0) differed significantly only in < 30% of both recordings on placebo and on digoxin. However, with increasing the distance between segments, the proportion of the significant differences between RR interval distributions increased more with placebo than with digoxin (P < 10(-300), Chi-square test). Paired data revealed larger differences between placebo and digoxin with increasing distance between segments. Thus in patients with paroxysmal AF, digoxin leads to more reproducible patterns of ventricular cycles that may be better tolerated clinically. PMID- 9744438 TI - Early benefit of implantable cardioverter defibrillator therapy in patients waiting for cardiac transplantation. AB - The ICD can effectively recognize and treat ventricular arrhythmias that can lead to sudden death. Sudden death is a major problem in patients awaiting heart transplantation. We reviewed our experience with the ICD in patients with malignant ventricular arrhythmias waiting for cardiac transplantation. Nineteen patients were included. Seventeen were men, mean age was 54 +/- 11 years (range 17-66) and the left ventricular ejection fraction was 22% +/- 10% (range 9%-46%). After a mean follow-up of 6 +/- 5 months (range 1-20 months), 17 patients reached heart transplantation. One patient died and the other is waiting for a transplant. Before transplantation 71% of patients received an appropriate discharge. The mean time to the first appropriate discharge was 2 +/- 2 months (range < 1-6 months), which was significantly shorter than the mean time to first discharge in the other patients (n = 182) receiving a defibrillator in our center (11 +/- 10 months; range 1-58 months) (P < 0.0004). In conclusion, cardiac transplantation candidates with life-threatening ventricular arrhythmias can effectively be protected against sudden arrhythmic death by ICD. These patients have a high incidence of appropriate shocks occurring very early after implantation. PMID- 9744439 TI - Prevention of atrial arrhythmias during DDD pacing by atrial overdrive. AB - We evaluated the effect of atrial overdrive on the incidence of atrial arrhythmias (AA) in 22 patients (67 +/- 9 years, 7 women, 15 men) with Chorus 6234 DDD pacemakers. Atrial overdrive was defined as a programmed paced rate 10 ppm faster than the mean ventricular rate stored for the last 24-hour period in the pacemaker memory. The protocol consisted of three phases of 1 month each. Phase I: observation after discontinuation of antiarrhythmic therapy. Phase II: arrhythmia analysis using the pacemaker memory after programming the lower rate to 55 ppm. The fallback function and histogram data were used to document the number and maximal duration of AA episodes as well as the total AA time in a month. Phase III: atrial overdrive. The mean ventricular heart rate was 65 +/- 4 beats/min before atrial overdrive versus 75 +/- 5 with atrial overdrive (P = 0.02). At the end of phase II, all patients presented with AA episodes (mean number per patient: 42 +/- 78 in one month). In phase III (with atrial overdrive), 14 (64.6%) patients had no recorded AA (group A). In the other eight patients with persistent AA episodes in phase III (group B), there was a significant reduction in the number of AA episodes (90 +/- 106 in phase II vs 38 +/- 87 in phase III; P = 0.01), their total duration (166 +/- 115 in phase II vs 92 +/- 134 hours in phase III; P = 0.03) and their maximal duration (121 +/- 103 in phase II vs 85 +/- 89 min; P = 0.04). Our short-term data suggest that atrial overdrive prevents or reduces AA episodes and demonstrate the feasibility and need of long-term studies to determine whether this benefit is sustained. PMID- 9744440 TI - Extensive variation in the signal amplitude of the atrial floating VDD pacing electrode. AB - The dependence of atrial signal amplitude on the site of the atrial sensing dipole of a single-pass lead was examined in 29 patients. The vertical location of the dipole was documented in supine fluoroscopy during quiescent and deep breathing and in upright chest roentgenogram, and was expressed as a proportion of the right atrial height. As the group average, the atrial signal amplitude was equal when tested in supine, sitting, standing, and right- or left-side positions in follow-up determinations. The signal amplitude varied markedly between postures, showing a coefficient of variation of 45% +/- 24% within the group. Coefficient of variation within the 6-month follow-up period in each tested position ranged from 31%-44%. Correlation between postures was weak (range of r = 0.53-0.81). Vertical location of the atrial dipole had no relationship to the signal amplitude. At least in one posture or test occasion the atrial signal amplitude was very low, < or = 0.35 mV in 20 patients, and below detection limit (0.25 mV) in 5 patients. Programmed to high sensitivity, atrial undersensing was rare in ambulatory electrocardiography, ranging from 0-9,000 missed atrial beats (0%-8%), with a median of 100 beats/24 hours. In conclusion, temporary variation in atrial signal amplitude is extensive. Despite occasionally measured large signal amplitudes atrial sensitivity in single lead VDD pacemakers should be programmed high, and if poor atrial tracking is suspected, other methods besides routine sensitivity testing should be adapted. PMID- 9744441 TI - Steady-state and dynamic behavior of ventricular repolarization and refractoriness in the dog: the effect of multiple cycle length changes and d sotalol administration. AB - In anesthetized dogs with chronic, complete AV block we studied the characteristics of ventricular repolarization and refractoriness. Therefore, we determined: (1) steady-state values of ventricular effective refractory period (VERP), action potential duration (APD), and stimulus T interval (STI) before and after d-sotalol treatment at various pacing cycle lengths (PCLs); and (2) the dynamics of VERP, APD, and STI before and after d-sotalol treatment after the abrupt PCL decreases. VERP, APD, and STI showed a normal frequency dependency. All three parameters increased significantly after d-sotalol administration. During steady-state and dynamic measurements, STI was always longer than APD and APD was always longer than VERP in an individual animal, irrespective of PCL and conditions. Standard deviations of steady-state and dynamic values indicated a considerable interindividual variation. However, the dynamics of VERP, APD, and STI after an abrupt decrease in PCL were highly correlated (linear regression analysis: r2 > or = 0.93). The best mathematical model to describe these dynamics was a bi-exponential model (r2 > or = 0.98) with a very short first and a much longer second time constant. We found that there was a very consistent relation between VERP, APD, and STI, not only during steady-state but also in the dynamic situation after various abrupt PCL decreases. This relation does not change after the administration of d-sotalol. Therefore, STI could be used to predict steady state and dynamic values of VERP and APD. Since STI can be made available online in implantable pacing systems this could lead to the development of new features in these devices. PMID- 9744442 TI - Worldwide clinical experience with a down-sized active can implantable cardioverter defibrillator in 162 consecutive patients. Worldwide 7221 ICD Investigators. AB - Treatment with an ICD is the first-line treatment for survivors of sudden cardiac death. More recently, evidence accumulates that prophylactic ICD therapy may be beneficial for selected subgroups of patients after myocardial infarction. Particularly for future studies on the value of prophylactic ICD therapy, downsized devices are needed to allow easy pectoral implantation with a single lead configuration and featuring extended memory capabilities. Accordingly, this study assesses the clinical performance of a downsized fourth-generation ICD in 162 consecutive patients. All devices could be successfully implanted pectorally, in 96% with a single lead configuration with a low defibrillation threshold of 10.6 +/- 5.2 J. During a 3-month follow-up, 26% of the patients received ICD therapy. Twenty percent had appropriate therapy for ventricular fibrillation (n = 9) and VT (n = 23), which was effective in all cases. Of the 450 episodes of VT, 426 were terminated by antitachycardia pacing. Fourteen patients (9%) had inappropriate ICD therapy mainly due to atrial fibrillation or sinus tachycardia, which could be reliably diagnosed by the ICD stored intracardiac electrograms. PMID- 9744443 TI - Interactions between pacemakers and security systems. AB - Electromagnetic fields arising from a variety of different sources have been shown to interfere with normal pacemaker function. This study evaluated the possible interactions between two modern security systems and different pacemaker types. Fifty-three patients (27 single chamber pacemakers, 25 dual chamber pacemakers) have been tested routinely for their pacemaker function. Thirty-eight patients presented with unipolar sensing and 15 with bipolar sensing. The patients were asked to walk through an installed security system, an antitheft device, and electromagnetic access device with different field strengths while a six-channel ECG monitored the patients. The pacemaker systems were first measured in their basic programmed modes, then the intervention frequency was changed to 100/min and, thereafter, the maximum sensitivity without T wave oversensing was added. In the security system with the highest field strength (2,700 mA/m), a pacemaker malfunction could be observed in 13% of the monitored patients. In one case, a pacemaker (VVIR) switched to ventricular safety pacing (VOO mode). In the security system with the lower field strength (1,600 mA/m) we found a pacemaker malfunction in 4% of the tested patients. In the antitheft device (50 mA/m), in the electromagnetic access device (300 mA/m), and in pacemaker systems with bipolar sensing, none of these dysfunctions were observed. Phantom programming as described previously did not occur in any of the systems. Persons who are often in the vicinity of security systems should be equipped with a bipolar pacemaker system. Our findings indicate that patients with pacemakers should avoid contact with security systems. PMID- 9744444 TI - The mean ventricular fibrillation cycle length: a potentially useful parameter for programming implantable cardioverter defibrillators. AB - In programming the implantable cardioverter defibrillator (ICD), the ventricular tachycardia (VT) detection cycle length (CL) is based on the CL of the documented tachycardia but the ventricular fibrillation (VF) detection CL is set arbitrarily. Appropriate programming of VF detection may not only reduce the incidence of inappropriate ICD shocks for non-VF rhythms but can also avoid the fatal underdetection of VF. The mean VFCL may provide a useful parameter for optimal ICD programming for VF detection if it is reproducible. This study examined the intrapatient reproducibility and interpatient variation of the mean VFCL in 30 ICD patients (25 men and 5 women, mean age 63 +/- 13 years). A total of 210 VF episodes (7 +/- 4 per patient, range 3-17) induced by T-wave shocks (166) or AC (44) at the ICD implant (30 patients) and the predischarge test (12 of 30 patients) were analyzed. The mean VFCL was calculated from the stored V-V intervals in the ICDs. Although the mean VFCL varied significantly from 171 +/- 6 to 263 +/- 11 ms (P < 0.01) among different patients, it was reproducible among different VF episodes in an individual patient (maximal variation 4-50 ms, P > 0.05). The mean VFCL was not significantly different between patients with and without antiarrhythmic drugs (210 +/- 32 vs 210 +/- 23 ms, P > 0.05) and was correlated with the ventricular effective refractory period (r = 0.5, P < 0.05). The mean VFCL varies greatly among different patients but remains reproducible in an individual patient, suggesting that the mean VFCL may serve as a reference for ICD programming of VF detection. PMID- 9744445 TI - Myocardial lactate extraction during repeated fibrillation/defibrillation episodes in defibrillator implantation testing. AB - Intraoperative testing with several fibrillation/defibrillation episodes (FDEs) is routinely performed during defibrillator implantation. Testing is considered safe even in patients with severe cardiac impairment, provided the recovery timespans and number of FDEs are adapted to the individual patient. Myocardial lactate extraction (MLE) was examined in two testing protocols. In 30 patients with coronary artery disease defibrillator implantations were performed under intravenous anesthesia. A percutaneous catheter was positioned into the coronary sinus (CS) underfluoroscopy. Two groups were randomly formed: group A (n = 20, mean number of FDEs: 4.2/patient) with 2 minutes waiting time between FDEs, and group B (n = 10, mean number of FDEs 4.1/patients) with 10 minutes between FDEs. Defibrillation pulses were released 15 seconds after T wave shock induced fibrillation. To estimate MLE, arterial and CS blood samples were collected before and after each FDE. After the last FDE, samples were obtained after 5, 10, and up to 20 minutes. In group A, MLE fell from a baseline value of 29.6% +/- 3.6% before the FDEs to 7.8% +/- 5.4% immediately after the episodes. MLE recovered to 27.2% +/- 6.5% within 1 minute and overshot to 35.6% +/- 5.8% within 5 minutes. In group B, MLE decreased from 37.6% +/- 7.5% to 15.1% +/- 8.1% immediately after each FDE and rose to its original value (33.6 +/- 7.8) within the 5-minute recovery period. MLE decreased immediately after each FDE, and recovered within 1 minute even in poor left ventricular function. For full MLE recovery a 2-minute wait between episodes is sufficient, if the total number of FDEs does not exceed four. PMID- 9744446 TI - Radiofrequency catheter ablation of atrioventricular nodal reentry tachycardia utilizing nonfluoroscopic electroanatomical mapping. AB - The advent of catheter ablation stimulated extensive research into anatomical localization of the pathways involved in atrioventricular nodal reentrant tachycardia (AVNRT). Conventional electrophysiological methods that attempt to correlate intracardiac electrograms with two-dimensional fluoroscopic anatomy are limited by the relative inaccuracy and poor reproducibility of this technique, and the requirement for high levels of radiation exposure. A new method of nonfluoroscopic electroanatomical mapping utilizes magnetic field sensing with a specialized catheter to construct three-dimensional electroanatomical endocardial maps of selected heart chambers with spatial resolution of < 1 mm. This system can be used in patients undergoing catheter ablation for AVNRT to create accurate maps of Koch's triangle and to guide application of radiofrequency energy. Initial experience in 14 patients suggests efficacy and safety comparable to conventional mapping and ablation techniques. Further evaluation may confirm the potential benefits of this system with respect to success rates, complications, procedure time, and radiation exposure. PMID- 9744447 TI - Electroanatomical mapping and ablation of the substrate supporting intraatrial reentrant tachycardia after palliation for complex congenital heart disease. AB - In patients with congenital heart disease who have undergone palliative surgical interventions postoperative arrhythmias frequently complicate the clinical course. Intraatrial reentrant tachycardias (IARTs) are one of the most common forms of postoperative arrhythmias in these patients and can lead to significant morbidity and even mortality. Drug therapy and/or antitachycardia pacing have been disappointing. Ablative therapy with radiofrequency energy offers a potential for cure for these patients but the conventional approach using multielectrode recordings and fluoroscopic guidance is technically difficult and provides limited success. Recent development of a novel nonfluoroscopic technology with electroanatomical mapping using the CARTO mapping/ablation system has shown promising results in defining the arrhythmia circuit, facilitating diagnosis, and guiding ablative therapy. Based on our preliminary experience, a systematic approach to postoperative IART using electroanatomical mapping is described. Further studies are needed to fully evaluate the impact of this new technology on the management and therapy of IART. PMID- 9744448 TI - Cardioversion after spontaneously terminated ventricular tachycardia: what is the mechanism? AB - We report on an 84-year-old patient with an ICD who received a shock after a spontaneously terminated nonsustained ventricular tachyarrhythmia. We discuss the possible mechanisms of cardioversion after conversion of ventricular tachycardia and interventions that may prevent this type of inappropriate shock. PMID- 9744449 TI - Access to biomaterials: a growing health policy concern. PMID- 9744450 TI - Idiopathic left ventricular tachycardia with block between purkinje potential and ventricular myocardium. AB - We performed radiofrequency current catheter ablation in a patient with idiopathic LV. While mapping the inferoapical LV septum during tachycardia, spontaneous termination of tachycardia was observed with block between Purkinje (P) potential and ventricular electrogram (P-V block). The cycle length of the tachycardia was associated with prolongation of P-P interval and P-V interval. P potential recording at this site was earliest and at very low amplitude during tachycardia. The radiofrequency current at this site was successful. These findings indicated that Purkinje fiber was a critical part of the tachycardia circuit. Ablation was successful at a site where both an earliest and low amplitude P potential was recorded during tachycardia, and where P-V block that was induced by catheter manipulation was observed during tachycardia. PMID- 9744451 TI - Pacing in distal left ventricular hypertrophic cardiomyopathy. AB - We report the case of an 85-year-old woman with distal LV hypertrophy resulting in an intraventricular pressure gradient and incomplete systolic emptying who benefited from permanent DDD pacing. Our experience based on this case suggests that permanent dual chamber pacing might be a safe and effective therapy also in symptomatic patients with the rare form of hypertrophic cardiomyopathy with left mid-ventricular obstruction resulting in incomplete emptying of the apical portion of the LV and a significant intraventricular pressure gradient that was not responding to pharmacological therapy. PMID- 9744452 TI - Unusual features of right and left idiopathic ventricular tachycardia abolished by radiofrequency catheter ablation. AB - Two unusual cases are presented with idiopathic right and left ventricular tachycardia (IVT) with intriguing clinical and electrophysiological characteristics. The first patient with a sustained IVT of right ventricular outflow tract origin, and an electrophysiological mechanism suggesting reentry, had been resuscitated from cardiac arrest. The second patient had an IVT with a left bundle branch block morphology, which originated from the basal-septal region of the left ventricle (left ventricular outflow tract tachycardia). Both patients were cured with radiofrequency catheter ablation, guided by endocardial activation sequence and pace mapping. PMID- 9744453 TI - Catheter ablation for ventricular tachycardia from a diverticulum at the right ventricular outflow tract. AB - A case is presented of a 73-year-old man with drug resistant ventricular tachycardia that originated from the right ventricular outflow tract. A right ventriculogram showed a diverticulum in the interventricular septum at the right ventricular outflow tract. Low energy radiofrequency catheter ablation within the diverticulum was performed successfully and safely. PMID- 9744454 TI - Recurrent polymorphic ventricular tachycardia complicating radiofrequency catheter ablation of the atrioventricular junction. AB - Recurrent ventricular tachycardia and ventricular fibrillation were observed immediately after RF ablation of the AV junction in a 64-year-old man. This arrhythmia was preceded by ventricular bigeminy and a long-short sequence. It was not associated with prolongation of the QT interval compared to baseline, and recurred 3 months later despite ventricular pacing at 90 beats/min. This is the first reported case of sustained ventricular arrhythmia complicating RF AV junction ablation despite rapid ventricular pacing, and recurring 3 months after discharge. It may explain the rare cases of sudden death complicating this procedure. PMID- 9744455 TI - Familial atrial standstill with coexistent atrial flutter. AB - A child with familial atrial standstill and a ventricular pacemaker had syncope due to atrial flutter that was treated by His-bundle ablation. Bradycardia protection alone may be insufficient in patients with atrial standstill. PMID- 9744456 TI - Parathyroidectomy via bilateral cervical exploration: a retrospective review of 866 cases. AB - INTRODUCTION: Parathyroidectomy via cervical exploration is an effective primary modality treatment for hyperparathyroidism, with cure rates of greater than 95%. We retrospectively reviewed 866 consecutive parathyroidectomies performed by a single surgeon between 1960 and 1997. We attempted to describe the polymorphic variation in multiglandular disease, the anatomic locations of pathologic glands, and the operative strategy and techniques which we believed were important to minimizing morbidity and maximizing curative success. METHODS: The cases of 329 males and 537 females (age, 1-88 years) were reviewed. There were 766 operations performed: primary hyperparathyroidism (713), tertiary hyperparathyroidism (100), reoperations (53). The strategy for primary exploration includes a bilateral neck exploration, early recurrent laryngeal nerve skeletonization, and identification of at least four glands. RESULTS: Normocalcemia was achieved in 98.2% of cases after initial cervical exploration. Persistent hypercalcemia occurred in 7 patients (<1%). Nine patients (1%) suffered persistent postoperative hypocalcemia. Unilateral recurrent laryngeal nerve injury occurred in two patients (<1%). Other perioperative complications included: reoperation for hematoma, repaired carotid artery injury, unexplained dysphagia, pneumothorax, deep venous thrombosis, and aspiration pneumonia. There were two mortalities (<1%) attributable to severe, comorbid disease. Ectopic glands were found in 120 cases. The frequency of glands at these sites were as follows: mediastinal (4.9%), intrathymic (8.4%), intrathyroid (6.7%), and retroesophageal/retrotracheal (3.5%). Thyroid resections provided diagnosis of concomitant thyroid carcinoma in 8.0% of resected patients. The pathology of patients with primary hyperparathyroidism (PHPT) consisted of single adenomas (77.2%), hyperplasia (21.0%), normal glands (1%), double adenomas (<1%), and parathyroid carcinoma (<1%). The distribution of adenomas was as follows: left upper, 25.3%; left lower, 27.3%; right upper, 26.8%; right lower, 20.6%. Hyperplastic glands were found in ectopic positions as follows: intrathymic (7.5%), intrathyroid (11.3%), mediastinal (2.5%), and retroesophageal/retrotracheal (0%). The average volume difference between the largest and smallest hyperplastic gland of each case was 1.80 + 4.40 cm3. Reoperations were performed upon 53 referred patients and 7 patients after failed exploration. Normocalcemia was attained in 98.3% of cases. Glandular pathology was identified in the previous operative field in 52 patients (86.7%). Adenomas were identified in 56.0% (n = 23) and hyperplasia in 39.0% (n = 16). CONCLUSIONS: In our series, we were able to attain normocalcemia in 98.2% of cases after initial cervical exploration. We believe that identification of four glands, an exhaustive search of ectopic sites, bilateral exploration, and liberal use of biopsy and intraoperative frozen section were essential to curative success. The pathologist should identify parathyroid tissue in the specimen and differentiate the "abnormal" from "normal" gland. Morphologic criteria alone cannot be used because of polymorphic variation in hyperplasia in which pathologic glands may appear normal. Early identification of the recurrent laryngeal nerve allows for a safer neck exploration by alerting the surgeon to the location and course of the nerve. A bilateral approach does not contribute increased morbidity from recurrent laryngeal nerve injury. PMID- 9744457 TI - Precisely defining high-risk operable head and neck tumors based on RTOG #85-03 and #88-24: targets for postoperative radiochemotherapy? AB - BACKGROUND: Local-regional recurrence of disease remains the major obstacle to cure of advanced head and neck cancers. METHODS: This investigation reviewed data derived from Radiation Therapy Oncology Group (RTOG) protocols #85-03 and #88-24 to identify characteristics of tumors that predicted local-regional recurrence of disease following surgery and postoperative radiotherapy (RT). RESULTS: The presence of tumor in two or more lymph nodes, and/or extracapsular spread of nodal disease, and/or microscopic-size tumor involvement of the surgical margins of resection imparts a high risk of local-regional (L-R) relapse. Our data also support the hypothesis that, following surgery, the concurrent addition of chemotherapy (CT) to RT may increase the likelihood of L-R control of disease for patients who have these high-risk characteristics. CONCLUSION: A prospective trial of surgery followed by concurrent RT and CT is warranted for patients who have high-risk characteristics found at surgery. PMID- 9744458 TI - Neo-adjuvant chemotherapy and supracricoid partial laryngectomy with cricohyoidopexy for advanced endolaryngeal carcinoma classified as T3-T4: 5-year oncologic results. AB - BACKGROUND: Historically, total laryngectomy with voice-prosthesis insertion and near-total laryngectomy were the surgical options advocated for advanced supraglottic and transglottic tumors classified as T3-T4. METHODS: The present retrospective study reviewed our experience with neo-adjuvant chemotherapy and supracricoid partial laryngectomy with cricohyoidopexy (SCPL-CHP) in a series of 60 patients with an isolated, untreated, advanced supraglottic/transglottic invasive squamous cell carcinoma classified as T3-T4. RESULTS: The Kaplan-Meier 5 year actuarial survival, local failure, nodal failure, and distant metastasis estimates were 72.7%, 8.3%, 9.2%, and 9.8%, respectively. Survival was significantly reduced in patients with nodal failure (p = .001) and distant metastasis (p = .007). Overall, a 91.7% laryngeal preservation rate and a 98.3% local control rate were achieved. CONCLUSION: Our report was a retrospective analysis and did not present a control group exclusively managed with SCPL-CHP. Therefore, we were unable to demonstrate that the use of neo-adjuvant chemotherapy prior to SCPL-CHP allowed for an increase in local control, laryngeal preservation, and survival. However, the use of neo-adjuvant chemotherapy allowed for remobilization of a fixed arytenoid cartilage in 10 patients who thus became amenable to SCPL-CHP. The key role of neo-adjuvant chemotherapy in this series was as a prognostic indicator for suitability for SCPL-CHP in the case of supraglottic-transglottic tumor with arytenoid cartilage fixation. Our data also supported the notion that SCPL-CHP is a valid alternative to total laryngectomy with voice prosthesis insertion and near-total laryngectomy in selected patients with a previously untreated supraglottic/transglottic invasive squamous cell carcinoma classified as T3-T4. Furthermore, the successful use (in terms of surgical outcome, laryngeal preservation, and survival) of SCPL CHP after neo-adjuvant chemotherapy suggested that laryngeal organ-preservation strategies, in advanced endolaryngeal transglottic and/or supraglottic invasive squamous cell carcinoma, should not be limited to the use of laryngeal radiotherapy after neo-adjuvant chemotherapy. PMID- 9744459 TI - Health states following head and neck cancer treatment: patient, health-care professional, and public perspectives. AB - BACKGROUND: This study investigated the assignment of preference values to health states which may follow head and neck cancer (HNC) treatment. Preference values for these health states were provided by HNC patients, HNC health-care providers, and a group of college students representing individuals with little knowledge of HNC. METHODS: A time trade-off technique was used by participants to assign preference values to four health states in the domains of appearance, eating, speech, breathing, pain, and work/social functioning. RESULTS: Patients' and health-care professionals' rank-ordered preference value scores for health states in appearance, breathing, eating, and speech were not significantly different (p < .05). These two groups differed significantly in ranking four of the eight pain and work/social functioning health states. Patients and students differed significantly in ranking 21 of the 24 health states (p < .05). CONCLUSIONS: Health-care professionals and patients had very similar perspectives regarding health states in the HNC-specific domains, indicating that these professionals appear to be a legitimate proxy for patients' attitudes in these domains. Healthcare professionals placed a significantly greater value on avoiding both pain and social confinement than did patients. Students, representing individuals naive regarding HNC, differed from patients and health-care professionals in their rankings of these health-state outcomes. PMID- 9744460 TI - Radiotherapy in the management of chemodectomas of the carotid body and glomus vagale. AB - BACKGROUND: Because only limited data are available pertaining to radiotherapy for chemodectomas of the carotid body and glomus vagale, we reviewed our experience. METHODS: Fifteen patients with 23 chemodectomas of either the carotid body or glomus vagale were treated with radiotherapy at the University of Florida between 1981 and 1995. Eighteen lesions were previously untreated. One patient had received prior radiotherapy at another institution and four patients had received prior surgery. RESULTS: The local control rate at 10 years, calculated by the Kaplan-Meier product-limit method, was 96% for the overall group of 23 lesions and 100% for the subset of 22 lesions without prior radiotherapy. The 10 year cause-specific survival rate was 89% for all 15 patients and 100% for the 14 patients who had received no prior radiotherapy. No patient experienced a significant complication secondary to irradiation. CONCLUSIONS: Irradiation offers a high probability of tumor control with relatively minimal risks for patients with chemodectomas of the carotid body and glomus vagale. PMID- 9744461 TI - Role of chest CT scanning in the management of patients presenting with head and neck cancer. AB - BACKGROUND: The detection of synchronous tumors, whether they be second primaries or distant metastases, in patients with head and neck carcinoma drastically affects prognosis and may alter management. Computerized tomographic (CT) scanning of the chest is an effective screening investigation in this group of patients, both in the detection of synchronous second primary tumors, the incidence of which in this study is 15%, and for accurate staging of metastatic pulmonary disease. The incidence of synchronous tumors in patients who are initially seen with head and neck squamous cell carcinoma (HNSCC) has been reported in large retrospective studies as being between 1% and 3%. These may be either second primary tumors or metastases, and the lung is the commonest site for both. METHODS: Eighty-one head and neck cancer patients (67 primary and 14 secondary referrals) treated at the Royal Liverpool University Hospital between 1994 and 1996 underwent CT scanning of the chest with ultrasound of the liver as part of their routine staging. The results were compared with standard chest x rays also performed in each patient. RESULTS: Fourteen patients had pulmonary tumors detected on the chest CT scan. In 67 patients, the scan was negative. Patients with negative scans tended not to have neck node metastases (64%), whereas patients with positive scans were much more likely to have neck node metastases with negative necks present in only 36% of patients. Where multivariate analysis was carried out, there was a correlation between neck node metastases and positive CT scans of the chest (estimate = 0.5755, standard error = 0.3066, chi2(1) = 6.73, p .047). The sensitivity of chest x-ray compared with CT scan was only 21 % and the specificity 99%. The positive predictive value of a chest x-ray was 75% and the negative predictive value 86%. Intra-abdominal lesions were detected in two patients, one in the liver and one in the adrenal gland. In the latter patient, this was an isolated lesion, but in the former, the chest scan was also positive. In the 67 patients, who were initially seen at the Royal Liverpool Hospital (primary referrals), the incidence of synchronous tumors was 15%. CONCLUSIONS: Synchronous tumors, whether they be second primary tumors or distant metastases, are more common in patients initially seen with head and neck cancer than is realized, their incidence being significantly higher in those patients with cervical metastases. Computerized tomographic scanning of the chest is a more effective screening investigation than chest x-ray in this group of patients and is now used routinely in our department prior to undertaking major head and neck surgery. PMID- 9744462 TI - High incidence of gastropharyngeal and gastroesophageal reflux after total laryngectomy. AB - BACKGROUND: Gastroesophageal reflux (GER) appears to be related to laryngeal carcinoma. Little is known about GER and gastropharyngeal reflux (GPR) in the laryngectomized patient. Therefore, GER and GPR were studied in laryngectomized patients. METHODS: In 11 patients, 24-hour double-probe pH monitoring was performed in an ambulant setting. An optic fiberscope was used for the accurate positioning of the proximal probe in the upper esophageal sphincter. RESULTS: In 9 of 11 patients pathologic GPR was found. Four of these 9 patients had reflux in upright and supine position, 5 patients had reflux only in upright position. CONCLUSIONS: A high incidence of GPR in laryngectomized patients was found. These results raise the question whether all laryngectomized patients should be investigated for reflux and in the presence of pathologic reflux findings should be treated with reflux prophylaxis. PMID- 9744463 TI - Complications of the myocutaneous platysma flap in intraoral reconstruction. AB - BACKGROUND: Pedicled myocutaneous flaps remain important tools in head and neck reconstruction. Evaluation of their complications are necessary to judge their merits. METHODS: From 1985 to 1995, 44 patients underwent a myocutaneous platysma flap reconstruction of the oral cavity or oropharynx. The following potential risk factors for complications were analyzed: age, sex, site of primary tumor, TNM stage, previous treatment, neoadjuvant chemotherapy, and type of operation. RESULTS: Flap-related complications occurred in 18 patients. Only one patient required reoperation for flap failure. Nineteen minor complications occurred in 17 patients. A significant increase in complications was seen in patients in which neoadjuvant chemotherapy was given. CONCLUSIONS: One flap failure was observed in our series. As the platysma flap has several advantages, it should be considered in the reconstruction of small intraoral defects. Contraindications are previous radiotherapy and surgery in the head and neck, neoadjuvant chemotherapy, nodal disease, and large defects. PMID- 9744464 TI - Partial laryngectomy as salvage surgery for radiation failures in T1-T2 laryngeal cancer. AB - BACKGROUND: Radiotherapy is the treatment of choice for early glottic carcinoma. Thirteen percent to 24% of patients require salvage surgery. To evaluate time of recurrence, site, and locoregional control, we retrospectively reviewed 29 patients treated from 1981 to 1996. METHODS: There were 28 men and 1 woman. Mean age was 63 years. Twenty were T1 (69%) and 9 were T2 (31%). Median time of recurrence was 14.5 months. In 14 patients (52%), a partial laryngectomy was done, and 13 patients had a total laryngectomy. Two refused surgery. RESULTS: One patient relapsed after salvage surgery. Five-year survival after salvage surgery was 92%, with no difference between partial and total laryngectomy (p = 0.2). CONCLUSIONS: Recurrences after failure to radiotherapy in T1-T2 glottic carcinoma could be salvaged with partial laryngectomy in 52% of patients, preserving laryngeal function, with adequate tumor control and acceptable morbidity. The selection of the surgical procedure is based on the tumor extension. PMID- 9744465 TI - Focal adhesion kinase expression in oral cancers. AB - BACKGROUND: Evidence suggests that focal adhesion kinase (FAK) is involved in the pathogenesis of certain cancers. Focal adhesion kinase is overexpressed in invasive and metastatic cancers of the breast, colon, thyroid, and prostate. The objective of this study was to determine the presence and cellular distribution of FAK in oral cancer and determine whether there is a difference in FAK expression in preinvasive and invasive oral cancers. METHODS: Immunohistochemistry was used to detect FAK expression in 20 archival oral cancer specimens. RESULTS: Focal adhesion kinase immunoreactivity was detected in all specimens that were examined. In tumors, there was an increase in the intensity of FAK staining and in the percentage of FAK-positive cells. Preinvasive tumors had populations of cancer cells which stained more intensely than neighboring cancer cells. CONCLUSIONS: Focal adhesion kinase expression appears to be increased in invasive and preinvasive oral cancers. It is speculated that enhanced expression of FAK may contribute to the aggressive phenotype of oral cancers. PMID- 9744466 TI - Expression of bcl-2-, p53, and Ki-67 and outcome of patients with primary nasopharyngeal carcinomas following DNA-damaging treatment. AB - BACKGROUND: Recent studies suggest that apoptosis is important in the cell death induced by treatment that damages deoxyribonucleic acid (DNA). We assessed the correlation between the expression of the apoptosis-related proteins, p53 and bcl 2, and the clinical outcome of patients with nasopharyngeal carcinomas (NPCs) who were treated with both DNA-damaging treatments. We also assessed the level of Ki 67, a marker of cell proliferation, in these tumors. METHODS: We evaluated statistically the relationships among the expression of p53, bcl-2, and Ki-67 and clinicopathologic factors, the sensitivity to radiation, the incidence of distant metastases, and survival. RESULTS: The group that was positive for p53 tended to be resistant to radiotherapy and to have a significantly poorer prognosis (p = .05). CONCLUSIONS: The enhanced expression of p53 may be a prognostic factor in patients with NPCs whose tumor is resistant to therapy that damages DNA. PMID- 9744467 TI - Nodular fasciitis in the parotid region of a child. AB - BACKGROUND: Nodular fasciitis is a common pathologic entity in the limbs of adults but rare in the head and neck of children. It is defined by the World Health Organization as a benign and probably reactive fibroblastic growth extending as a solitary nodule from superficial fascia into subcutaneous tissue. Treatment is local excision, and recurrence is rare. METHOD: Case Report RESULTS: A 3.5-year-old boy was initially seen with a 1-year history of gradually enlarging but otherwise asymptomatic right facial mass. On examination, a firm nodule was palpable anterior to the right ear, and facial movement was symmetrical. Computed tomography showed a rounded, well-defined solid mass continuous with the parotid fascia. The patient underwent superficial parotidectomy without complication. The pathology was reported as nodular fasciitis, and the child has had no clinical recurrence over 2 years. CONCLUSION: Benign lesions in this region in children may present similarly to malignancies but require much more-conservative treatment. PMID- 9744468 TI - Primary malignant teratoma of the thyroid gland: report and discussion of two cases. AB - BACKGROUND: Teratoma of the thyroid in adults is a rare neoplasm and is usually seen in young females. Most of the thyroid teratoma are malignant. The tumor appears as a dominant mass in the thyroid gland and is often associated with local lymph node involvement. We describe our experience of primary malignant teratoma of the thyroid in two young women who were treated with aggressive chemotherapy and surgical intervention. METHODS: Medical records of two patients treated between 1993 and 1995 were reviewed. Both patients were women (36 years old and 34 years old). The diagnosis of primary malignant teratoma of the thyroid was made on the basis of clinical, radiographic, and microscopic findings. Patients were treated with aggressive combination chemotherapy consisting of alternating regimens similar to those used for high-volume germ-cell tumor patients at the University of Texas M. D. Anderson Cancer Center. RESULTS: One of the patients demonstrated a partial response to chemotherapy and underwent postchemotherapy surgery for removal of residual disease. The other patient had a complete response to chemotherapy. Both patients are alive and disease free for 32- and 26-plus months. CONCLUSIONS: Primary malignant teratoma of the thyroid is sensitive to combination chemotherapy. It appears that the treatment strategy offered-aggressive induction chemotherapy with planned surgery for removal of residual disease, similar to that for patients with testicular tumors-has the potential to provide a durable, complete remission. PMID- 9744469 TI - Ameloblastic carcinoma of the mandible. AB - BACKGROUND: Ameloblastic carcinoma is a rare, aggressive odontogenic neoplasm of the jaws in which the epithelial cells exhibit cytologic features of recognizable ameloblastoma and malignancy. Cases with metastasis have been infrequently reported. METHODS: A case of a 64-year-old white woman with mandibular ameloblastic carcinoma with documented distant metastasis is presented. The patient's presenting symptoms included facial asymmetry of the right jaw over 2 months and the development of moderate trismus. Clinical manifestations, pathology, treatment, and biologic behavior are discussed. The nomenclature and classification of odontogenic carcinomas are reviewed, including entities that should be considered in the differential diagnosis. RESULTS: The patient underwent surgical resection consisting of mandibulectomy, parotidectomy, and modified radical neck dissection followed by radiation to both necks and tumor bed. Postsurgically, the patient developed pulmonary metastasis at 11 months and expired with widespread metastatic disease at 28 months. CONCLUSIONS: This case demonstrated an unusual behavior pattern in that local recurrence and regional metastasis did not occur. Distant metastasis occurred despite apparent adequate control of the primary mandibular tumor. The ameloblastic carcinoma is a highly malignant neoplasm which requires aggressive therapy. Prognosis is poor. Further reporting of ameloblastic carcinoma is encouraged. PMID- 9744470 TI - Free composite myo-osseous flap with serratus anterior and rib: indications in head and neck reconstruction. PMID- 9744471 TI - Single nucleotide polymorphism hunting in cyberspace. AB - Large-scale sequencing of human cDNA and genomic DNA libraries has produced a large collection of sequence data in public databases. To date, >900,000 human expressed sequence tag (EST) sequences and >80,000,000 bases of genomic DNA sequence have been deposited in Genbank. This ever-expanding data set is a rich source of gene-associated and anonymous single nucleotide polymorphisms (SNPs). DNA sequence variations can be found by comparing the sequences of redundant ESTs and by comparing sequences from overlapping genomic clones. Initial studies have shown that, with proper computer screening, informative SNP markers can be developed from these DNA databases in an efficient and cost-effective manner. Complete public access to these databases will allow individual investigators to add biological value to the human sequence data generated by large-scale sequencing centers. PMID- 9744472 TI - Mutations of the human E-cadherin (CDH1) gene. AB - The cell-cell adhesion molecule E-cadherin is well known to act as a strong invasion suppressor in experimental tumor cell systems. Frequent inactivating mutations have been identified for the E-cadherin gene (CDH1) in diffuse gastric cancers and lobular breast cancers. To date, 69 somatic mutations have been reported comprising, in addition to few missense mutations, mainly splice site mutations and truncation mutations caused by insertions, deletions, and nonsense mutations. Interestingly, there is a major difference in mutation type between diffuse gastric and infiltrative lobular breast cancers. In diffuse gastric tumors, the predominant defects are exon skippings, which cause in-frame deletions. By contrast, most mutations found in infiltrating lobular breast cancers are out-of-frame mutations, which are predicted to yield secreted truncated E-cadherin fragments. In most cases, these mutations do occur in combination with loss of heterozygosity (LOH) of the wild-type allele. Inactivating germline mutations of E-cadherin were recently reported for families with early-onset diffuse gastric cancer. Also, at the early stages of sporadic lobular breast and diffuse gastric cancers, E-cadherin mutations were detected, suggesting loss of growth control by such mutations and defining E-cadherin as a true tumor suppressor for these particular tumor types. PMID- 9744473 TI - The R496H mutation of arylsulfatase A does not cause metachromatic leukodystrophy. AB - Deficiency of arylsulfatase A (ARSA) enzyme activity causes metachromatic leukodystrophy (MLD). A number of ARSA gene mutations responsible for MLD have been identified. Recently, the R496H mutation of ARSA was proposed to be a cause of MLD (Draghia et al., 1997). We have investigated the R496H mutation and found this mutation at a relatively high frequency in an African American population (f=0.09, n=61 subjects). The ARSA enzyme activity in subjects with and without the R496H mutation was determined and found to be normal. It is therefore concluded that the R496H mutation of ARSA does not negatively influence the activity of ARSA and is not a cause of MLD. PMID- 9744474 TI - Prevalence of glucocerebrosidase mutations in the Israeli Ashkenazi Jewish population. AB - Gaucher disease is the most prevalent inherited disease among Ashkenazi Jews. It is very heterogeneous due to a large number of mutations within the glucocerebrosidase gene, whose impaired activity is the cause for this disease. Aiming at determining Gaucher carrier frequency among the Ashkenazi Jewish population in Israel, 1,208 individuals were molecularly diagnosed for six mutations known to occur among Ashkenazi Jewish Gaucher patients, using the newly developed Pronto Gaucher kit. The following mutations were tested: N370S, 84GG, IVS2+1, D409H, L444P, and V394L. Molecular testing of these mutations also allows identification of the recTL allele. The results indicated that Gaucher carrier frequency is 1:17 within the tested population. The prevalence of N370S carriers is 1:17.5. This implies that approximately 1:1225 Ashkenazi Jews will be homozygous for the N370S mutation. Actually, in our study of 1,208 individuals one was found to be homozygous for the N370S mutation. The actual number of known Ashkenazi Jewish Gaucher patients with this genotype is much lower than that expected according to the frequency of the N370S mutation, suggesting a low penetrance of this mutation. Results of loading experiments in cells homozygous for the N370S mutation, as well as cells homozygous for the L444P and the D409H mutations, exemplified this phenomenon. PMID- 9744475 TI - Characterization of N93S, I312T, and A333P missense mutations in two Japanese families with mitochondrial acetoacetyl-CoA thiolase deficiency. AB - Mitochondrial acetoacetyl-CoA thiolase (T2) deficiency is an inborn error of ketone body and isoleucine catabolisms. Japanese patients, GK01 and GK19, were found to be compound heterozygotes of 149delC and A333P, and N93S and I312T, respectively. The latter three missense mutations were individually characterized by analyses of transient expression of the cDNAs and heat stability. A333P and I312T subunits showed aberrant electrophoretic mobility on SDS-PAGE. T2 protein was destabilized by A333P and existed as an insoluble form in the mitochondria. I312T mutation also destabilized T2 protein; however, some T2 protein was retained in soluble form and reduced residual activity was apparent. N93S mutation did not change the heat stability of T2 activity and the reduced residual activity was retained, however a considerable amount was observed in an insoluble form. The effects of mutations were interpreted based on a tertiary structural model of a subunit of the human T2. This model was constructed from the X-ray crystal structure of the homologous peroxisomal 3-ketoacyl-CoA thiolase of Saccharomyces cerevisiae. On the basis of this model, the positions of Ala333 and Ile312 were far from the active site and the mutations would be expected to destabilize the tertiary structure of T2 subunit. By contrast, Asn93 is located near the active site and may function to maintain a local loop structure. The mutation of Asn93 could directly disrupt disposition of the active site. PMID- 9744476 TI - Prevalence of Lithuanian mutation among St. Petersburg Jews with familial hypercholesterolemia. AB - We used polymerase chain reaction-single-strand conformation polymorphism (PCR SSCP) analysis to detect LDL receptor gene defects in the St. Petersburg population. We have found a deltaG197 mutation in several patients of Jewish origin. The mutation named is shown to be responsible for one-third (7/23) of familial hypercholesterolemia (FH) cases in St. Petersburg Jews and absent in patients of Russian descent. The prevalence of a deltaG197 mutation in St. Petersburg Jews is consistent with its origin in Lithuania or Poland. The deltaG197 mutation can be easily detected in polyacrylamide minigels because of formation of specific heteroduplexes during PCR with DNA of heterozygous patients. Taken together with high prevalence of the mutation in St. Petersburg Jews, this observation provides an opportunity for DNA diagnostics of FH in this ethnic group. PMID- 9744477 TI - Identification of novel L1CAM mutations using fluorescence-assisted mismatch analysis. AB - The L1CAM gene, which is located in Xq28 and codes for a neuronal cell adhesion molecule, is involved in three distinct conditions: HSAS (hydrocephalus-stenosis of the aqueduct of Sylvius), MASA (mental retardation, aphasia, shuffling gait, adductus thumbs), and SPG1 (spastic paraplegia). Molecular analysis of the L1CAM gene is labor-intensive because of the size of the coding region, which is fragmented in numerous exons, and because of the great allelic heterogeneity and distribution of the mutations. The FAMA (fluorescent assisted mismatch analysis) method combines the excellent sensitivity of the chemical cleavage method for scanning PCR fragments larger than 1 kb and the power of automated DNA sequencers. In order to optimize this method for L1CAM, we divided the gene into nine genomic fragments, each including three to four exons. These fragments were PCR-amplified using nine sets of primers containing additional rare universal sequences. A second-stage PCR, per formed with the two dye-labeled universal primers, allowed us to generate 1-kb-labeled fragments, which were then submitted to the chemical cleavage analysis. Among 12 French families with HSAS and/or MASA, we identified nine distinct L1CAM mutations, seven of which were novel, and an intronic variation. This study demonstrates that FAMA allows rapid and reliable detection of mutations in the L1CAM gene and thus represents one of the most appropriate methods to provide diagnosis for accurate genetic counseling in families with HSAS, MASA, or SPG1. PMID- 9744478 TI - Dihydropteridine reductase deficiency: physical structure of the QDPR gene, identification of two new mutations and genotype-phenotype correlations. AB - Dihydropteridine reductase (DHPR) is an enzyme involved in recycling of tetrahydrobiopterin (BH4), the cofactor of the aromatic amino acid hydroxylases. Its deficiency is characterized by hyperphenylalaninemia due to the secondary defect of phenylalanine hydroxylase and depletion of the neurotransmitters dopamine and serotonin, whose syntheses are controlled by tryptophan and tyrosine hydroxylases. The DHPR cDNA has been cloned and mapped on 4p15.3. In the present study we report the genomic structure of the DHPR gene (QDPR). This gene includes seven exons within a range of 84-564 bp; the corresponding introns are flanked by canonic splice junctions. We also present a panel of PCR primers complementary to intronic sequences that greatly facilitates amplification of the gene and provides a genomic DNA approach for mutation detection. We have used this approach to study six patients with DHPR deficiency. Four known mutations (G23D, H158Y, IVS5G+ 1A, R221X) and two new mutations (Y150C and G218ins9bp) were found. The Y150C mutation was found in compound heterozygosity with G23D, a mutation always associated with a severe phenotype in homozygous patients. This patient has an intermediate phenotype (good response to monotherapy with BH4). The mutant enzyme for Y150C was expressed in an E. coli system. Comparison of its kinetic parameters with those of the G23D mutant enzyme showed that it is not as effective as the wild-type enzyme, but is more active than the G23D mutant. This patient's intermediate phenotype is thus due to the mild DHPR mutation Y150C. Correlations between genotypes and phenotypes were also found for the other mutations. PMID- 9744479 TI - Mutation 1091delC is highly prevalent in Spanish Sanfilippo syndrome type A patients. AB - The gene resposible for Sanfilippo syndrome type A, a lysosomal disorder caused by deficiency of sulfamidase, was recently cloned and more than 40 mutations were identified. This paper presents the mutation analysis and clinical findings in 11 Spanish patients in whom 19 of the 22 mutant alleles have been identified. This is the first report on mutations in Spanish Sanfilippo A patients. Seven different mutations were found, four of which (Q85R, R206P, A354P, and L386R) were not previously described. Mutation 1091delC was the most prevalent, accounting for nearly one-half of the mutated alleles, while mutations R245H and R74C were not found. Haplotype analysis suggests a founder effect as the cause of the high frequency of 1091delC in this population. PMID- 9744480 TI - A phosphoglycerate kinase mutant (PGK Herlev; D285V) in a Danish patient with isolated chronic hemolytic anemia: mechanism of mutation and structure-function relationships. AB - Phosphoglycerate kinase (PGK) is a X-linked enzyme that plays a key role in the glycolytic pathway. Twelve different variants have already been reported. We describe a new PGK variant, PGK Herlev (Asp 285-->Val), in a 69-year-old Danish patient with isolated chronic hemolysis but who had no neurological or muscular disorders. The description of the mutation is based upon PCR amplification of specific regions of the PGK gene, followed by direct sequencing. Although observed in a male patient, this mutated X-linked gene is expressed partially, i.e., both normal and substituted nucleotides are present at the same position in a ratio of approximately 1:9. The most likely explanation for this observation is based on the occurrence of a somatic mutation of the PGK gene. The relationship of structure to function in PGK Herlev, as well as in all known variants, was examined by the use of a computer model based on the known spatial structure of the yeast and horse enzymes. Such an approach can be generalized to any other protein that has been crystallized and for which x-ray diffraction data are available in a species closely related to man. PMID- 9744481 TI - Leptin receptor immunoreactivity is associated with the Golgi apparatus of hypothalamic neurons and glial cells. AB - Leptin has emerged as a major peripheral hormone, controlling central mechanisms of metabolism and related autonomic and endocrine functions. In the regulation of hypothalamic neurones, leptin is suggested to affect different second messenger systems and transcription regulating factors. The present study reports the predominant localization of leptin receptor immunoreactivity in the cis and trans cisternae of the Golgi apparatus in hypothalamic neuronal and glial cells. In these hypothalamic cells, translocation of leptin receptor immunoreactivity from the Golgi apparatus to the perikaryal membrane, nucleus or to the cytoplasm was not apparent after manipulation of the metabolic state either by fasting or suppression of the thyroid axis. On the other hand, leptin receptor immunoreactivity was associated with the perikaryal membrane of neurones in other parts of the central nervous system, including the dentate gyrus and the cingulate cortex. These data indicate an extremely high turnover of leptin receptors in hypothalamic target sites, but also raise the possibility that leptin may interact with the Golgi apparatus-related mechanisms to alter intracellular mechanisms. PMID- 9744482 TI - The ovine melatonin-related receptor: cloning and preliminary distribution and binding studies. AB - A melatonin-related receptor was cloned from an ovine genomic library. The sequenced gene has a similar structure to that of the melatonin receptor gene family and consists of two exons separated by an intron of approximately 3 kb. Exon 1 and exon 2 of the ovine melatonin-related receptor encode a protein of 575 amino acids which is 73.8% homologous to the human melatonin-related receptor and shows 40.9% homology with the ovine Mel1a melatonin receptor. COS-7 cells transiently expressing ovine melatonin-related receptors did not bind 2 [125]iodomelatonin or 3H-melatonin. Reverse transcription-polymerase chain reaction (RT-PCR) and in situ hybridization studies revealed expression of the ovine melatonin-related receptor in the hypothalamus, pituitary, retina and retinal pigment epithelium. Furthermore, expression of the ovine melatonin related receptor is shown to be coincident with Mel1a and 2-[125I]iodomelatonin binding in the pituitary and serotonin N-acetyl transferase (arylalkylamine N acetyl transferase, AANAT) expression in the retina. Expression patterns and similarity with the melatonin receptor gene family suggest a role for this novel G protein-coupled receptor in control and regulation of endocrine function and retinal physiology. PMID- 9744484 TI - Effect of NMDA receptor antagonist MK-801 on light-induced Fos expression in the suprachiasmatic nuclei and on melatonin production in the Syrian hamster. AB - In mammals, circadian rhythms generated by the suprachiasmatic nuclei (SCN) are daily synchronized by a light-dark cycle. Photic information is transmitted to the SCN mainly through the direct retinohypothalamic tract, the neurotransmitters involved being excitatory amino acids. It is also commonly accepted that photoperiodic information coming from the retina via the SCN is transduced by the pineal into a nocturnal signal, i.e. melatonin production. Light exposure at night induces (1) an inhibition of melatonin synthesis and (2) an expression of c fos in numerous cells of SCN. To determine the role of the NMDA receptor in these effects, we treated Syrian hamsters with ip injections of MK-801, a noncompetitive NMDA receptor antagonist. Several subpopulations of light sensitive cells in the SCN are affected by MK-801. According to previous studies, MK-801 inhibits light-induced Fos immunoreactivity mainly in the most ventral part of the SCN. However, we observed that numerous other cells are still activated by light. When light is applied in the middle of the night, MK-801 pretreatment does not reduce Fos-ir in the dorsal SCN. At the beginning of the night, labeled cells in this part of the nucleus appear even more numerous after MK-801. We also found that MK-801 fails to reduce the light-induced inhibition of melatonin synthesis. Moreover, in control animals, which received no light stimulation, ip injection of MK-801 induces by itself a dose-dependent inhibition of melatonin production. PMID- 9744483 TI - Corticotropin releasing factor mRNA expression in the hypothalamic paraventricular nucleus and the central nucleus of the amygdala is modulated by repeated acute stress in the immature rat. AB - Age-appropriate acute stress, such as cold exposure, provokes the secretion of corticotropin releasing factor (CRF) from the hypothalamus, leading to a robust increase of plasma corticosterone in the immature rat. This activation of the hypothalamic-pituitary-adrenal system is accompanied by a stress-induced increase of steady-state CRF-mRNA expression in the hypothalamic paraventricular nucleus (PVN). In the current study, we analysed changes in CRF-mRNA expression in the PVN and the central nucleus of the amygdala (ACe) in the immature rat in response to a single episode of cold stress and three repeated exposures to this same stressor. CRF-mRNA expression in the PVN increased after a single, but not repeated exposures to cold stress, while repeated acute stress increased the content of the CRF peptide in the anterior hypothalamus. In the ACe, repeated episodes of cold stress resulted in increased expression of CRF-mRNA. These findings indicate a differential regulation of CRF gene expression in the PVN and ACe of the immature rat by single and repeated acute stress. PMID- 9744485 TI - N-methyl-D-aspartic acid stimulation of vasopressin release: role in osmotic regulation and modulation by gonadal steroids. AB - Previous experiments demonstrated that excitatory amino acids participate in the osmotic regulation of vasopressin secretion, but the specific involvement of N methyl-D-aspartic acid (NMDA) receptors was not evaluated. This was demonstrated in the present studies. NMDA stimulated vasopressin release from perifused explants of the hypothalamo-neurohypophyseal system (HNS), and osmotic stimulation of vasopressin release was inhibited by MK-801 (10 microM) and AP5 (100 microM) NMDA receptor antagonists. The effective concentration of NMDA was dependent upon the Mg2+ concentration of the perifusate with stimulation observed at 1 microM NMDA in Mg2+-replete compared with 5 microM in low-Mg2+ medium. Previous experiments also demonstrated that estradiol and dihydrotestosterone (DHT) inhibited osmotically stimulated vasopressin secretion, and a nongenomic mechanism of action was suggested by the ability of steroids conjugated to bovine serum albumin to replicate the effect. Experiments were performed to explore the potential role of NMDA receptors in this mechanism. Estradiol (50 pg/ml) and DHT (3 ng/ml) inhibited NMDA stimulated vasopressin release in perifused HNS explants. These results suggest a role of NMDA receptors in the mediation of vasopressin secretion in osmotically stimulated release. Furthermore, estradiol and DHT may exert their inhibitory effect on osmotically stimulated vasopressin release via the NMDA receptor. PMID- 9744486 TI - Identification of alpha1B adrenergic receptor protein in gonadotropin releasing hormone neurones of the female rat. AB - Noradrenaline is an important neurotransmitter which regulates GnRH release from the median eminence in the female rat during both basal GnRH secretion and the preovulatory or steroid hormone-induced GnRH-mediated LH surge. However, it is not clear at which sites in the brain this predominantly stimulatory influence is exerted nor is it known which adrenergic receptor subtype(s) mediate(s) the effects of noradrenaline. In order to determine if the GnRH neurones in the septum-diagonal band-preoptic area and/or their axon terminals in the median eminence are direct targets for noradrenaline, immunohistochemical triple labelling studies were conducted to localize simultaneously GnRH peptide, dopamine-beta-hydroxylase and alpha1B adrenergic receptor protein. The results show that about 80% of all GnRH neurones examined contained patches of immunoreactive alpha1B adrenergic receptor protein at or near the plasma membrane and that some of these alpha1B adrenergic receptors were adjacent to dopamine beta-hydroxylase containing axons. The GnRH neurones which did not contain alpha1B adrenergic receptors were preferentially located in the rostral portion of the septum and diagonal band while all GnRH neurones in the caudal septum, diagonal band and in the preoptic area expressed alpha1B adrenergic receptors. In the median eminence, a few alpha1B adrenergic receptor patches were seen in the external layer and these receptors were only rarely observed to be associated with GnRH containing axon terminals. The results suggest that the effects of noradrenaline on GnRH release are, at least in part, mediated by the activation of alpha1B adrenergic receptors which are located on most GnRH perikarya while the median eminence is not a likely site at which GnRH release is regulated by alpha1B adrenergic receptors. PMID- 9744487 TI - Axons containing the growth associated protein GAP-43 specifically target rat corticotrophs following adrenalectomy. AB - An extensive network of nerve fibers immunoreactive for the neuronal growth associated protein GAP-43 (GAP-43-IR) is present within the anterior pituitary (AP) of the rat, and the density of these fibers has been reported to increase 4 days after adrenalectomy (ADX). In the present study, we employed confocal dual label immunofluorescence microscopy to determine whether GAP-43-IR fibers are specifically associated with corticotrophs at various intervals after ADX. A dramatic increase in the density of GAP-43-IR was apparent 4 days after ADX, and this increase was sustained at 7 and 14 days post-ADX. The percentage of corticotrophs in apparent contact with GAP-43-IR axons was 87% at 4 days after ADX and 92% at 14 days. In addition, fewer than 15% of GAP-43-IR terminals were associated with cells other than corticotrophs in either group. This highly specific targeting of corticotrophs during a period in which these cells are undergoing both hypertrophy and hyperplasia indicates that axonal sprouting is occurring in response to ADX. While the less intense GAP-43-IR in the AP of intact rats precluded precise quantitative analysis, the majority of corticotrophs also appeared to be selectively innervated in these animals. The observations that GAP-43-IR axons selectively contact corticotrophs, and that both the specificity and thoroughness of innervation are maintained by targeted growth of GAP-43-IR axons following ADX, strongly suggest that these fibers are of functional significance. PMID- 9744488 TI - Seasonal neuroendocrine rhythms in the male Siberian hamster persist after monosodium glutamate-induced lesions of the arcuate nucleus in the neonatal period. AB - The aim of these experiments was to examine the role of the arcuate nucleus in the control of seasonal cycles of body weight, feed intake, moulting and reproduction in the Siberian hamster. The arcuate nucleus has previously been implicated as a central site where systemic feedback signals (e.g. leptin) might act to regulate feed intake and body weight, so it was predicted that hamsters with lesions of this structure would be unable to display the inhibitory effects of short days on these parameters. In the first series of studies, lesions that destroyed approximately 80% of the cells in the arcuate nucleus were produced by treating hamsters neonatally with monosodium glutamate (MSG; 4 mg/g body weight sc), and vehicle- and MSG-treated males were raised from birth in long days (LD) or short days (SD). In hamsters raised in LD, the initial gain in body weight and testicular growth were significantly reduced by MSG treatment, however, growth rate and testis weight were still significantly greater than in vehicle- or MSG treated hamsters raised in SD. In the second study, hamsters treated neonatally with vehicle or MSG were raised in LD for 8 weeks and, subsequently, approximately half in each group were transferred to SD for 18 weeks. As expected, vehicle-treated hamsters showed a characteristic decline in body weight when exposed to SD, while those remaining in LD continued to increase body weight. Feed intake decreased in parallel with the decline in body weight in SD, a complete moult to the white winter pelage occurred by 16 weeks in SD, and testicular regression occurred. Responses to SD also occurred in the MSG-treated hamsters: body weight decreased in SD but increased in their lesioned litter mates remaining in LD, and feed intake paralleled body weight changes in these groups. The moult to winter pelage was significantly retarded in MSG-treated hamsters transferred to SD. The testes were completely regressed in sham- and MSG treated hamsters exposed to SD, whereas testes weights in MSG-treated hamsters maintained in LD were intermediate between those in vehicle-treated hamsters in SD and LD. Thus, despite initial effects on growth, the MSG-treated hamsters bearing substantial lesions of the arcuate nucleus were able to show appropriate responses to photoperiod, although not always of the same magnitude as the unlesioned controls. We conclude that feedback mechanisms operating via the arcuate nucleus are not the major regulators of seasonal cycles of body weight, feed intake, pelage and reproduction. PMID- 9744489 TI - Immunohistochemical distribution of oestrogen receptor and luteinizing hormone B subunit in the ovine pituitary gland during foetal development. AB - The presence of oestrogen receptor in the developing hypothalamo-hypophyseal system is an essential prerequisite for the development of sex-steroid feedback on gonadotrophin secretion. We have used dual immunocytochemistry to examine the ontogeny and regional distribution of oestrogen receptor and LHbeta subunit in the ovine pituitary gland during foetal development. At day 65 gestation (term= 145 days) oestrogen receptor and LH/ immunopositive cells are found in a small region at the base of the anterior pituitary gland, and also in a band immediately adjacent to the neurointermediate lobe. By day 100 gestation there was a significant increase in the number of immunopositive LHbeta cells accounting for around 12% of the total cell population, and these were widely distributed throughout the anterior pituitary gland. There was also a significant increase in the proportion of gonadotrophs which contain oestrogen receptor compared with day 65. By day 130 gestation the percentage of LH containing cells had declined to around 7% of the total population, but the proportion which also contained oestrogen receptor remained the same. There were no differences in the numbers or distribution of cells containing LH or oestrogen receptors between male and female foetuses, at any age. These data describing a parallel change in the number of oestrogen receptors and LHbeta containing cells in the pituitary gland throughout gestation suggest that the development of pituitary sex-steroid feedback is not solely dependent on changes in the numbers of oestrogen receptor containing cells alone. PMID- 9744490 TI - Sleep endocrine effects of megestrol acetate in healthy men. AB - Synthetic and naturally occurring steroids exert a variety of neural effects that include modulation of nocturnal sleep. The present study focuses on the effect of progesterone receptor (PR) activation on the nocturnal sleep electroencephalogram (EEG) in male volunteers. As a PR ligand, the synthetic progesterone megestrol was used, which has the advantage over progesterone in that it is not metabolized into other steroid compounds which could cloud the progesterone-mediated effects through their own neuroactive properties. Nine healthy male volunteers were investigated in a prospective single-blind randomized study design. They received either placebo tablets or megestrol acetate dosages of 160, 320 or 480 mg at 14.00 h and 19.00 h. Blood samples were drawn half-hourly from 22.00 h until 07.00 h. After 320 mg megestrol, plasma adrenocorticotropin secretion was lower and growth hormone secretion was higher than after 160 mg and 480 mg megestrol or placebo. Similarly, the reduction in the relative amount of rapid eye movement sleep was most pronounced after 320 mg. Thus, progesterone receptor activation, as reflected by the sleep EEG and associated pituitary hormone secretion, follows a nonlinear U-shape dose dependency of a well-defined PR ligand, which may explain the unresolved inconsistencies of neuroendocrine progesterone effects to date. Moreover, employing a CV1 cell line, contransfected with a human glucocorticoid receptor expression vector and a reporter gene-based detection system for transcriptional activity, revealed that a PR agonist such as megestrol may also activate glucocorticoid receptors. This may account for some of the neuroendocrine effects of megestrol and other progestins. PMID- 9744491 TI - Evidence that an HLA-DQA1-DQB1 haplotype influences susceptibility to childhood common acute lymphoblastic leukaemia in boys provides further support for an infection-related aetiology. AB - Comparison of DQA1 and DQB1 alleles in 60 children with common acute lymphoblastic leukaemia (c-ALL) and 78 newborn infant control subjects revealed that male but not female patients had a higher frequency of DQA1*0101/*0104 and DQB1*0501 than appropriate control subjects. The results suggest a male associated susceptibility haplotype in c-ALL and supports an infectious aetiology. PMID- 9744492 TI - Variation in the survival of women with breast cancer in Scotland. The Scottish Breast Cancer Focus Group and The Scottish Cancer Therapy Network. AB - We have investigated factors influencing the survival of women with early breast cancer in Scotland. In a retrospective study, clinical, treatment and 'service' factors, e.g. surgical case load, deprivation and geographical area (health board of first treatment) were recorded from hospital records. A total of 2148 women with invasive breast cancer diagnosed in 1987 were identified from the Scottish Cancer Registry, of whom 1619 without metastases at diagnosis underwent surgery as part of their primary treatment. In a multivariate analysis, clinical factors (age, clinical stage, pathological tumour size, node status and oestrogen receptor status) all influenced survival. After allowing for these clinical factors, surgical case load and deprivation did not have statistically significant effects on survival. By contrast, health board did affect survival. This was explained in part by the selection of patients for surgery. There appeared, however, to be a residual effect that may be related to differences in the use of adjuvant systemic treatment among the different health boards. We conclude that, in Scotland, geographical variation in both surgical and non surgical treatment has a greater effect on variability in survival for women with breast cancer than surgical case load and deprivation. PMID- 9744493 TI - Effect of tamoxifen and transdermal hormone replacement therapy on cardiovascular risk factors in a prevention trial. Italian Chemoprevention Group. AB - The combination of tamoxifen and transdermal hormone replacement therapy (HRT) may potentially reduce risks and side-effects of either agent, but an adverse interaction could attenuate their beneficial effects. We assessed the effects of their combination on cardiovascular risk factors within a prevention trial of tamoxifen. Baseline and 12-month measurements of total, low-density lipoprotein (LDL)- and high-density lipoprotein (HDL)-cholesterol, platelets and white blood cells were obtained in the following four groups: tamoxifen (n = 1117), placebo (n = 1112), tamoxifen and HRT (n = 68), placebo and HRT (n = 87). The analysis was further extended to women who were on HRT at randomization but discontinued it during the 12-month intervention period (n = 33 on tamoxifen and n = 35 on placebo) and to women who were not on HRT but started it during intervention (n = 36 in both arms of the study). Compared with small changes in the placebo group, tamoxifen was associated with changes in total, LDL- and HDL-cholesterol of approximately -9%, -19% and +0.2% in continuous HRT users compared with -9%, -14% and -0.8% in never HRT users. Similarly, there was no interaction on platelet count. In contrast, the decrease in total and LDL-cholesterol levels induced by tamoxifen was blunted by two-thirds in women who started HRT while on tamoxifen (P = 0.051 for the interaction term). We conclude that the beneficial effects of tamoxifen on cardiovascular risk factors are unchanged in current HRT users, whereas they may be attenuated in women who start transdermal HRT while on tamoxifen. Whereas a trial of tamoxifen in women already on transdermal HRT is warranted, prescription of HRT during tamoxifen may attenuate its activity. PMID- 9744494 TI - Preservation of macronutrient preferences in cancer anorexia. AB - Indirect evidence suggests that cancer anorexia is associated with specific aversions to macronutrients. To investigate this, patients with cancer anorexia and hospitalized control subjects devised 3-day menus comprising foods that they wished to eat. These foods were then provided for 3 days and the intakes of each food carefully measured. As expected, patients with cancer anorexia consumed substantially less energy than hospitalized control subjects (6.0 +/- 0.9 MJ vs 9.5 +/- 0.5 MJ, P < 0.001). However, macronutrient composition was consistently maintained in the patients with cancer anorexia. These data argue against cancer anorexia representing a state of macronutrient aversion. PMID- 9744495 TI - Mutations of adenomatous polyposis coli (APC) gene are uncommon in sporadic desmoid tumours. AB - Desmoids are locally aggressive, non-metastasizing soft-tissue tumours, whose aetiology is still unclear. In patients affected with familial adenomatous polyposis (FAP), the incidence of desmoids is much higher than in the general population. The APC gene, which is responsible for FAP, is involved in the development of desmoids associated with this syndrome. In this study 16 sporadic and four FAP-related desmoids were analysed in order to investigate the possible involvement of APC in non-syndromic cases also. The 5' end (exons 1-11) and the coding portion of exon 15 of APC were screened using the in vitro synthesized protein assay (IVSP). Exons 5, 6, 8-14, and a region of exon 15 spanning codons 1036-1634 were investigated by single-strand conformation polymorphism (SSCP) analysis. APC germline mutations were identified in all FAP patients, but not in sporadic cases. Somatic mutations were found in three FAP-associated desmoids (75%) and two sporadic tumours (12.5%). In one of the latter cases, both alleles were affected. These findings indicate a limited role of the gene in the development of desmoid tumours outside FAP. PMID- 9744496 TI - The role of nitric oxide in bacillus Calmette-Guerin mediated anti-tumour effects in human bladder cancer. AB - Bacillus Calmette-Guerin (BCG) has been used for many years to treat cancer of the urinary bladder. It constitutes effective intravesical therapy of carcinoma in situ and recurrent superficial bladder cancer. Although the mechanism of action is unknown, most evidence suggests an immune-mediated mechanism. BCG treatment is known to increase cytokine production in the urinary bladder. As cytokines may induce nitric oxide synthase (NOS) activity and as nitric oxide (NO) exerts cytotoxic effects on tumour cells, we investigated the role of NO in BCG-mediated anti-tumour activity. Here we demonstrate a marked induction of both calcium-dependent and calcium-independent NOS activity in the human urinary bladder after BCG treatment. The presence of NOS in the urothelial cells was also demonstrated by the use of immunohistochemistry. Furthermore, patients treated with BCG showed a 30 times higher production of gaseous NO as measured in the urinary bladder by chemiluminescence. Finally, NO donors exerted cytotoxic effects on bladder cancer cell lines. These findings suggest that NO synthesis may be an important mechanism in BCG-mediated anti-tumour therapy. PMID- 9744497 TI - Cytotoxic effect of the cyclosporin PSC 833 in multidrug-resistant leukaemia cells with increased expression of P-glycoprotein. AB - Multidrug resistance (MDR) to anti-cancer agents is frequently associated with overexpression of the drug efflux transporter P-glycoprotein (Pgp) in cancer cells, ensuing drug expulsion and maintenance of tolerable intracellular levels of certain cytotoxic drugs. Pgp may also be present in normal tissue, providing protection against toxic substances, but the physiological role of Pgp is not fully understood. Recently, it was shown that Pgp also takes part in the transport of certain growth-regulating cytokines (Drach et al, 1996; Raghu et al, 1996). Therefore, we studied the effect of the highly potent Pgp inhibitor PSC 833 on proliferation of three pairs of MDR and parental human cell lines (HB8065 hepatoma cells, KG1a and K562 leukaemia cells). The MDR phenotypes were characterized by Pgp overexpression, which was demonstrated by flow cytometry using the anti-Pgp antibody MRK16. Electronic cell counting of 72-96 h cultures revealed a dose-dependent antiproliferative effect of PSC 833 in the resistant KG1a/200 and K562/150 cells. The half-maximal growth inhibitory concentrations (GI50) were 0.2 microM and 0.7 microM respectively. Exposure to PSC 833 induced cell death by apoptosis in both cell types, as revealed by flow cytometry and detection of 3'-hydroxy ends of DNA (the result of DNA fragmentation associated with apoptosis), by terminal transferase-mediated dUTP-biotin nick end-labelling (TUNEL). Similar effects were not found in the hepatoma cell lines or the parental leukaemia lines. These results demonstrated a discriminating cytotoxicity of PSC 833 in two human leukaemia MDR variants, representing a possible therapeutic indication which warrants consideration during the ongoing clinical evaluation of this drug. PMID- 9744498 TI - Aberrant transcripts of the FHIT gene are expressed in normal and leukaemic haemopoietic cells. AB - Deletions and apparent transcriptional abnormalities of the FHIT gene at 3p14.2 have recently been reported in a wide variety of solid tumours. To determine whether lesions of this gene also occur in leukaemia, we have analysed a total of 97 patients (chronic myeloid leukaemia, CML, in chronic phase or blast crisis, n = 71; de novo acute leukaemia, n = 26) and 16 normal individuals. Intact FHIT transcripts from all cases were amplified using RT-PCR. In addition, smaller size bands that were less intense than the full-length products were amplified from several samples from patients with leukaemia and also from normal leucocytes. Sequencing of the small products revealed that they were derived from FHIT transcripts lacking whole exons. Using single-strand conformation polymorphism analysis, no mutations in the coding sequence were detected in any patient. Furthermore, loss of heterozygosity was not seen in any of 36 informative patients at D3S1300 or D3S1481, markers located within the FHIT locus. We conclude that the FHIT gene and other uncharacterized tumour-suppressor genes at 3p14.2 are unlikely to be involved in the pathogenesis of acute leukaemia or progression of CML from chronic phase to blast crisis. Moreover, low-abundance FHIT transcripts that lack whole exons are not specific to malignant cells and should not be taken as evidence of an abnormality in the absence of demonstrable genomic DNA lesions. PMID- 9744500 TI - Sequential loss of heterozygosity in the progression of squamous cell carcinoma of the lung. AB - Radiographically occult bronchogenic squamous cell carcinomas are early lung cancers that localize mainly in the bronchial wall, and are thought to be a good model for investigating genetic alterations through lung cancer progression. In order to elucidate sequential genetic changes in lung cancers, we analysed the incidence of allelic losses on chromosome regions 2q33, 3p21, 5q21, 7q31, 9p21 and 17p13 for 40 cases of radiographically occult bronchogenic squamous-cell carcinomas and 40 cases of advanced lung cancers microdissected. In this study we used eight microsatellite dinucleotide polymorphic markers. Frequent loss of heterozygosity (LOH) was observed on 3p21 (53%), 5q21 (44%) and 17p13 (61%) in roentgenographically occult bronchogenic squamous cell carcinomas. 2q, 7q and 9p were lost less frequently in both roentgenographically occult bronchogenic squamous cell carcinomas and advanced lung cancers. These results suggest that several tumour-suppressor genes are associated with lung cancer progression and that genetic changes on 3p21, 5q21 and 17p13 are early events. PMID- 9744499 TI - Estimations of intra- and extracellular volume and pH by 31P magnetic resonance spectroscopy: effect of therapy on RIF-1 tumours. AB - Quantification of metabolite or drug concentrations in living tissues requires determination of intra- and extracellular volumes. This study demonstrates how this can be achieved non-invasively by 31P magnetic resonance spectroscopy (MRS) employing dimethyl methylphosphonate (DMMP) as a marker of total water space, 3 aminopropylphosphonate (3-APP) as a marker of extracellular space and P and 3-APP as markers of intracellular pH (pH) and extracellular pH (pHe) respectively. The MRS measurements of the tumour volumes were validated by classic radiolabelling methods using 3H2O and [14C]inulin as markers of total and extracellular space respectively. The extracellular volume fraction measured by radiolabelling of RIF 1 tumours was 23 +/- 0.83% (mean +/- s.e.m. n = 9), not significantly different (P > 0.1) from that found by MRS (27 +/- 2.9%, n = 9, London, and 35 +/- 6.7, n = 14, Baltimore). In untreated RIF-1 tumours, pH was about 0.2 units higher than pHe (P < 0.01). 5-Fluorouracil (5FU) treatment (165 mg kg(-1)) caused no significant changes in either pHe or per cent extracellular volume. However significant increases in pH, 48 h after treatment (P < 0.01) correlated with decreased tumour size and improved bioenergetic status [NTP/inorganic phosphate (Pi) ratio]. This study shows the feasibility of an MR method (verified by a 'gold standard') for studying the effects of drug treatment on intra- and extracellular spaces and pH in solid tumours in vivo. PMID- 9744501 TI - Comparative analysis of interleukin 15 and interleukin 2 for induction of killer activity and of type 2 cytokine production by mononuclear cells from lung cancer patients. AB - Interleukin (IL) 15 is a novel cytokine with IL-2-like activity. In this study, we examined the effect of IL-15 on induction of non major histocompatibility complex (MHC)-restricted killer activity and of type 2 cytokine production by peripheral blood and pleural mononuclear cells (MNCs), from 34 lung cancer patients and 20 control subjects. IL-15 induced significant killer activity in blood MNCs from lung cancer patients as well as control subjects against a small cell lung cancer cell line (SBC-3). Effective killer induction by IL-15 was observed even in blood MNCs and pleural MNCs from the site of tumour growth in advanced lung cancer patients. IL-12 had an additive effect with a suboptimal dose of IL-15 in induction of killer activity. In the case of MNCs from lung cancer patients, IL-10 production was more prominent when cells were incubated with IL-2 than with IL-15. IL-5 production was observed in MNCs from lung cancer patients stimulated with IL-2, but not with IL-15. These observations suggest that IL-15, by virtue of its lesser induction of type 2 cytokine, may be a better candidate than IL-2 for lung cancer immunotherapy. PMID- 9744502 TI - Autonomous proliferation and bcl-2 expression involving haematopoietic cells in patients with myelodysplastic syndrome. AB - In this work, we investigated the autonomous proliferation, bcl-2 expression and number of apoptotic cells in the bone marrow of patients with confirmed diagnosis of myelodysplastic syndromes (MDS). Normal bone marrow cells obtained from donors of the Clinical Hospital of this university were used as a control. The autonomous proliferation, evaluated by clonal culture without exogenous growth factor, and the number of apoptotic cells in bone marrow kept for 10 days in liquid cultures at 37 degrees C and 5% carbon dioxide, were significantly greater in MDS patients than in control subjects (P = 0.001, Wilcoxon). However, bcl-2 expression, measured by immunocytochemistry, was significantly lower in MDS patients than in normal individuals (P = 0.002, Wilcoxon). These results suggest that the high proliferation activity in MDS patients may be counteracted by the high level of medullar cell death, which might be related to the lower bcl-2 expression. PMID- 9744503 TI - Induction of apoptosis in human tumour xenografts after oral administration of uracil and tegafur to nude mice bearing tumours. AB - Various types of anti-neoplastic agents induce apoptosis in vitro, but less is known of the role of this mode of cell death in tumours treated in vivo. We examined the induction of apoptosis by oral anti-neoplastic agents, tegafur and uracil (UFT, a combined preparation of 1 mol tegafur and 4 mol uracil), and the relationship of effects on tumour growth. Seven different human gastrointestinal tumour xenografts were transplanted into nude mice, including two colon adenocarcinomas (KM20C and Col-1), three gastric carcinomas (SC-6, St-40 and 4 1ST) and two pancreatic carcinomas (PAN-4 and PAN-12), followed by oral administration of UFT (24 mg kg(-1) day(-1)) for 9 days. The percentage of apoptotic cells in each tumour was scored in histological sections, chronologically, using a molecular biological-histochemical system and growth inhibition was examined in each tumour. A significant growth inhibition by UFT was observed for all tumours, except PAN-12. In KM20C and SC-6, growth inhibition rates were 61.7% and 60.6% respectively. Quantitative assay for apoptosis showed a remarkable induction of apoptosis in KM20C (4.2%) and SC-6 (3.5%), which were relatively sensitive to UFT. In addition, KM20C and SC-6 showed a higher incidence of spontaneous apoptosis. In five other tumours, which responded to a lesser extent than KM20C and SC-6, UFT altered little the changes in apoptosis (less than 2%) and spontaneous apoptosis was relatively low. Thus, tumours with a higher apoptosis induced by UFT had a higher response to UFT. Apoptosis observed in tumours might serve as a predictor of a preferable response to UFT. PMID- 9744504 TI - In vitro invasion of small-cell lung cancer cell lines correlates with expression of epidermal growth factor receptor. AB - Formation of metastasis is a multistep process involving attachment to the basement membrane, local proteolysis and migration into surrounding tissues, lymph or bloodstream. In the present study, we have analysed the correlation between in vitro invasion and presence of the epidermal growth factor receptor (EGFR) in a panel of 21 small-cell lung cancer (SCLC) cell lines. We have previously reported that ten of these cell lines expressed EGFR protein detected by radioreceptor and affinity labelling assays. In 11 small-cell lung cancer (SCLC) cell lines, EGFR mRNA was detected by Northern blot analysis. In vitro invasion in a Boyden chamber assay was found in all EGFR-positive cell lines, whereas no invasion was detected in the EGFR-negative cell lines. Quantification of the in vitro invasion in 12 selected SCLC cell lines demonstrated that, in the EGFR-positive cell lines, between 5% and 16% of the cells added to the upper chamber were able to traverse the Matrigel membrane. Expression of several matrix metalloproteases (MMP), of tissue inhibitor of MMP (TIMP) and of cathepsin B was evaluated by immunoprecipitation, Western blot analysis and reverse transcriptase polymerase chain reaction (RT-PCR). However, in vitro invasive SCLC cell lines could not be distinguished from non-invasive cell lines based on the expression pattern of these molecules. In six SCLC cell lines, in vitro invasion was also determined in the presence of the EGFR-neutralizing monoclonal antibody mAb528. The addition of this antibody resulted in a significant reduction of the in vitro invasion in three selected EGFR-positive cell lines. Our results show that only EGFR-positive SCLC cell lines had the in vitro invasive phenotype, and it is therefore suggested that the EGFR might play an important role for the invasion potential of SCLC cell lines. PMID- 9744505 TI - Induction of cell death by stimulation of protein kinase C in human epithelial cells expressing a mutant ras oncogene: a potential therapeutic target. AB - Ras oncogene activation is a key genetic event in several types of human cancer, making its signal pathways an ideal target for novel therapies. We previously showed that expression of mutant ras sensitizes human thyroid epithelial cells to induction of cell death by treatment with phorbol 12-myristate 13-acetate (PMA) and other phorbol esters. We have now investigated further the nature and mechanism of this cell death using both primary and cell line models. The cytotoxic effect of PMA could be blocked by bisindolylmaleimide (GF 109203X), a well-characterized inhibitor of c and n protein kinase C (PKC) isoforms, and by prior down-regulation of PKC, indicating that it is mediated by acute stimulation, rather than down-regulation. Western analysis identified two candidate isoforms--alpha and epsilon--both of which showed PMA-induced subcellular translocation, either or both of which may be necessary for PMA induced cell death. Immunofluorescence showed that PMA induced a rapid nuclear translocation of p42 MAP kinase of similar magnitude in the presence or absence of mutant ras expression. Cell death exhibited the microscopic features (chromatin condensation, TdT labelling) and DNA fragmentation typical of apoptosis but after a surprising lag (4 days). Taken together with recent models of ras-modulated apoptosis, our data suggest that activation of the MAPK pathway by PMA tips the balance of pro- and anti-apoptotic signals generated by ras in favour of apoptosis. The high frequency of ras mutations in some cancers, such as cancer of the pancreas, which are refractory to conventional chemotherapy, together with the potential for stimulating PKC by cell-permeant pharmacological agents, makes this an attractive therapeutic approach. PMID- 9744506 TI - Phase I study of temozolomide in paediatric patients with advanced cancer. United Kingdom Children's Cancer Study Group. AB - A phase I study of temozolomide administered orally once a day, on 5 consecutive days, between 500 and 1200 mg m(-2) per 28-day cycle was performed. Children were stratified according to prior craniospinal irradiation or nitrosourea therapy. Sixteen of 20 patients who had not received prior craniospinal irradiation or nitrosourea therapy were evaluable. Myelosuppression was dose limiting, with Common Toxicity Criteria (CTC) grade 4 thrombocytopenia occurring in one of six patients receiving 1000 mg m(-2) per cycle, and two of four patients treated at 1200 mg m(-2) per cycle. Therefore, the maximum-tolerated dose (MTD) was 1000 mg m(-2) per cycle. The MTD was not defined for children with prior craniospinal irradiation because of poor recruitment. Plasma pharmacokinetic analyses showed temozolomide to be rapidly absorbed and eliminated, with linear increases in peak plasma concentrations and systemic exposure with increasing dose. Responses (CR and PR) were seen in two out of five patients with high-grade astrocytomas, and one patient had stable disease. One of ten patients with diffuse intrinsic brain stem glioma achieved a long-term partial response, and a further two patients had stable disease. Therefore, the dose recommended for phase II studies in patients who have not received prior craniospinal irradiation or nitrosoureas is 1000 mg m(-2) per cycle. Further evaluation in diffuse intrinsic brain stem gliomas and other high-grade astrocytomas is warranted. PMID- 9744507 TI - What is the effect of adjusting epirubicin doses for body surface area? AB - Doses of cytotoxic drugs are routinely adjusted according to body surface area. We have evaluated this practice in 32 women with advanced breast cancer treated with single-agent epirubicin 12.5-120 mg m(-2). Epirubicin and its metabolites were measured by high-performance liquid chromatography (HPLC). Unadjusted plasma clearance was calculated from dose in mg, and adjusted clearance from dose in mg m(-2). Unadjusted clearance did not correlate with surface area, height, weight, per cent ideal body weight or body mass index. There was no difference in the coefficient of variation (CV) of adjusted and unadjusted clearance (39.4% and 37.7% respectively). The AUC that would have resulted from giving an unadjusted dose was calculated. This predicted AUC was accurate, unbiased and had the same CV as the actual AUC. Similarly, in 11 patients an analysis of actual and predicted neutropenia confirmed that unadjusted dosing would have had no significant effect on the pattern of myelosuppression. Normalization of epirubicin dosage according to surface area appears not to reduce either pharmacokinetic or pharmacodynamic variability. PMID- 9744508 TI - Measurement and evaluation of serum anti-p53 antibody levels in patients with lung cancer at its initial presentation: a prospective study. AB - Anti-p53 antibodies in sera are known to be products of the host immune response to mutated p53 protein, and are present in some patients with various types of cancer. In this study, we measured serum anti-p53 antibody levels in 52 patients with lung cancer and 63 normal volunteers to determine the relationship between anti-p53 antibody level and clinical features of lung cancer patients. Anti-p53 antibody level was measured by an enzyme-linked immunosorbent assay and expressed as an anti-p53 antibody index, defined as the ratio of absorption of serum sample to that of p53-positive serum. The median anti-p53 antibody index was 6.6 for lung cancer patients, and higher than that in normal volunteers (1.7) (P = 0.0000). For lung cancer patients, significant differences in index levels were found by histology (4.3, n = 25, adenocarcinoma vs 8.7, n = 18, squamous cell carcinoma vs 64.8, n = 2, large-cell carcinoma vs 9.8, n = 7, small-cell carcinoma; P = 0.0109). High anti-p53 antibody index levels were observed for both large-cell carcinoma and small-cell carcinoma. When the cut-off level was set at 7.2, determined using the twice 95% specificity level for normal volunteers, the sensitivities of anti-p53 antibodies were 46.1% for all lung cancers, 28.0% for adenocarcinoma, 55.6% for squamous cell carcinoma, 100% for large-cell carcinoma and 71.4% for small-cell carcinoma. However, there were no significant differences in index level by gender, age, smoking index, presence of previous or concomitant cancer or disease stage. Multivariate analysis using a logistic regression model demonstrated that histological type of tumour was a dominant factor associated with elevation of anti-p53 antibody index level (P = 0.0184). These findings suggest that serum anti-p53 antibody index level might be independent of tumour burden and the presence of previous or concomitant cancer in our series of lung cancer patients, but is clearly strongly correlated with tumour histological type. PMID- 9744509 TI - Effective treatment of advanced breast cancer with vinorelbine, 5-fluorouracil and l-leucovorin plus human granulocyte colony-stimulating factor. AB - A phase II trial was performed to investigate the efficacy and tolerance of vinorelbine (VNB), 5-fluorouracil (5-FU), l-leucovorin (LLV) and recombinant human granulocyte colony-stimulating factor (G-CSF) in advanced breast cancer. Between August 1994 and October 1996, 53 patients entered this trial. Thirty seven patients were previously untreated and 16 patients had failed previous palliative chemotherapy with (n = 12) or without anthracyclines (n = 4). Therapy consisted of VNB 40 mg m(-2) diluted in 250 ml of saline infused over 30 min on days 1 and 14 and LLV 100 mg m(-2) administered by intravenous bolus injection and 5-FU 400 mg m(-2) diluted in 500 ml of saline infused over 2 h, both given on days 1-5 every 4 weeks. G-CSF was administered at 5 microg kg(-1) day(-1) subcutaneously on days 6-10 during each cycle. Treatment was continued in cases of response or stable disease until a total of six courses were completed. The overall response rate was 59% for chemotherapeutically naive patients (95% confidence interval 42-75%), including five complete responses (CR; 13%) and 17 partial responses (PR; 46%); ten patients (27%) had stable disease (SD) and only five (14%) progressed (PD). Second-line chemotherapy with this regimen resulted in 3/16 (19%) objective remissions, but nine patients had SD and four had PD. The median time to progression was 10.5 months (range 2-23) in previously untreated patients and 7.0 months (range 2-19) in those who had failed prior chemotherapy. After a median follow-up time of 14 months, 29 patients (55%) are still alive with metastatic disease; median survival has not been reached yet. The dose limiting toxicity was myelosuppression: WHO grade III and IV neutropenia occurred in 15 (28%) and four patients (8%), and was complicated by septicaemia in two; grade III anaemia and thrombocytopenia were noted in four (8%) and three (6%) patients respectively. Severe (WHO grade 3) non-haematological toxicities included stomatitis in 6% and nausea/vomiting and alopecia in 2% each. Our data suggest that the combination of vinorelbine, 5-fluorouracil and l-leucovorin plus G-CSF is an effective first line regimen for treatment of advanced breast cancer. Overall toxicity was modest, with myelosuppression being the dose-limiting side effect. Other severe adverse reactions were uncommon. PMID- 9744510 TI - Biochemical basis of 5-aminolaevulinic acid-induced protoporphyrin IX accumulation: a study in patients with (pre)malignant lesions of the oesophagus. AB - Administration of 5-aminolaevulinic acid (ALA) leads to porphyrin accumulation in malignant and premalignant tissues, and ALA is used as a prodrug in photodynamic therapy (PDT). To understand the mechanism of porphyrin accumulation after the administration of ALA and to investigate whether ALA-induced protoporphyrin IX might be a suitable photosensitizer in Barrett's oesophagus and adenocarcinoma, we determined the activities of porphobilinogen deaminase (PBG-D) and ferrochelatase (FC) in various malignant and premalignant as well as in normal tissues of the human oesophagus. A PDT power index for ALA-induced porphyrin accumulation, the ratio of PBG-D to FC normalized for the normal squamous epithelium of the oesophagus, was calculated to evaluate intertissue variation in the ability to accumulate porphyrins. In malignant and premalignant tissue a twofold increased PBG-D activity and a marginally increased FC activity was seen compared with normal squamous epithelium. A significantly increased PDT power index in Barrett's epithelium and adenocarcinoma was found. Our results suggest that, after the administration of ALA, porphyrins will accumulate in a greater amount in Barrett's epithelium and adenocarcinoma of the oesophagus because of an imbalance between PBG-D and FC activities. The PDT power index here defined might be a useful indicative parameter for predicting the susceptibility of these tissues to ALA-PDT. PMID- 9744511 TI - Randomized study of chemotherapy and surgery versus radiotherapy for stage IIIA non-small-cell lung cancer: a National Cancer Institute of Canada Clinical Trials Group Study. AB - Thirty-one patients with stage IIIA (N2) non-small-cell lung cancer were randomized to receive radiotherapy alone or chemotherapy with cisplatin and vinblastine followed by surgery. Response rates to induction chemotherapy and radiotherapy were 50% and 53.3% respectively. Complete surgical resection was possible for 62.5% of patients. Median survival times were 16.2 and 18.7 months for radiotherapy alone and chemotherapy-surgery respectively (P = Ns), with no long-term improvement in survival seen with combined-modality treatment. PMID- 9744512 TI - Factors affecting baseline quality of life in two international adjuvant breast cancer trials. International Breast Cancer Study Group (IBCSG). AB - Quality of life (QL) is used to assess treatments in clinical trials but may be influenced by other factors. We analysed the impact of biomedical, sociodemographic and cultural factors on baseline QL indicators in two International Breast Cancer Study Group trials. Patients with stage II breast cancer were randomized within 6 weeks of primary surgery to various adjuvant treatments. They were asked to assess five indicators of QL at baseline. QL forms were available for 1231 (83%) of the 1475 premenopausal and 989 (82%) of the 1212 post-menopausal patients, who were from nine countries and spoke seven languages. Culture (defined as language/country groups) had a statistically significant impact on baseline QL measures. Premenopausal patients with poor prognostic factors showed a tendency to report worse QL, with oestrogen receptor status as an independent predictor for mood (P = 0.0005). Older post-menopausal patients reported better emotional wellbeing (P = 0.002), mood (P = 0.002), and less effort to cope (P = 0.0009) compared with younger post-menopausal patients. Co morbidity, type of surgery, treatment assignment and sociodemographic factors showed a statistically significant impact in post-menopausal patients only. Cultural and biomedical factors influenced baseline QL and should be considered when evaluating the impact of treatment on QL in international breast cancer clinical trials. PMID- 9744513 TI - Recent improvement in survival of breast cancer patients in Saarland, Germany. AB - A new method for more timely monitoring of cancer patient survival was employed to assess progress in 5-year survival of breast cancer patients in Saarland, Germany, between 1980-84 and 1990-94. Five-year relative survival gradually increased from 68.8% to 73.5%. Improvements were most pronounced among age groups 50-59 and 60-69. The latter had the highest 5-year relative survival (77.1%) in 1990-94. PMID- 9744514 TI - Differential gene induction by glucocorticoid and progesterone receptors. AB - The question of how glucocorticoid, progesterone, androgen, and mineralocorticoid receptors can generate distinct patterns of gene expression despite similar, if not identical, DNA sequence recognition properties is a central question in steroid biology. This study addresses the hypothesis that glucocorticoid and progesterone receptors can differentially utilize the promoter context created by a series of receptor recognition sites. This hypothesis predicts that for different receptors an individual (often suboptimal) binding site contributes to the overall response element activity to a different degree. Therefore, mutations to an individual binding site of a multipartite hormone response element may differentially affect the response to different steroids. This study tests this hypothesis by introducing mutations into a receptor recognition site that is part of a hormone responsive promoter derived from the mouse mammary tumor virus. We find that weakening one site of a multipartite element differentially affects glucocorticoid and progestin signaling both in fibroblasts and in mammary carcinoma cells. A similar test was done in a simplified promoter context comprised only of a TATA box and a pair of receptor recognition sites. Mutations introduced into one of these sites impaired glucocorticoid response more than the progestin response. However, high receptor expression can partially overcome the effect of some target site mutations. Thus both receptor expression levels and the inherent ability to utilize the context created by a multipartite response element contribute to quantitative differences in the response to receptors that share DNA sequence recognition properties. PMID- 9744515 TI - Overview of a rational approach to design type I 17beta-hydroxysteroid dehydrogenase inhibitors without estrogenic activity: chemical synthesis and biological evaluation. AB - Hormone-sensitive diseases such as breast cancer are health problems of major importance in North America and Europe. Endocrine therapies using antiestrogens for the treatment and the prevention of breast cancer are presently under clinical trials. Antiestrogens are drugs that compete with estrogens for the estrogen receptor without activating the transcription of estrogen-sensitive genes. However, an optimal blockade of estrogen action could ideally be achieved by a dual-action compound that would antagonize the estrogen receptor and inhibit the biosynthesis of estradiol. Type I 17beta-hydroxysteroid dehydrogenase (17beta HSD) was chosen as a key steroidogenic target enzyme to inhibit the formation of estradiol, which is the most potent estrogen. This article describes a rational approach that could lead to the development of compounds that exhibit both actions. The chemical syntheses of estradiol derivatives bearing a bromoalkyl and a bromoalkylamide side chain at the 16alpha-position are summarized. Two parameters were studied for biological evaluation of our synthetic inhibitors: (1) the inhibition of estrone reduction into estradiol by type I 17beta-HSD, and (2) the proliferative/antiproliferative cell assays performed on the estrogen sensitive ZR-75-1 breast tumor cell line. First, the substitution of the 16alpha position of estradiol by bromoalkyl side chain led to potent inhibitors of type I 17beta-HSD, but the estrogenic activity remained. Secondly, an alkylamide functionality at the 16alpha- or 7alpha-position of estradiol cannot abolish the estrogenic activity without affecting considerably the inhibitory potency on type I 17beta-HSD. In conclusion, the best dual-action inhibitor synthesized showed an IC50 of 13 +/- 1 microM for type I 17beta-HSD, while displaying antiestrogenic activity at 1.0 microM. Despite the fact that we did not obtain an ideal dual action blocker, we have optimized several structural parameters providing important structure-activity relationship. PMID- 9744516 TI - Expression of vitamin D receptor (VDR) in HL-60 cells is differentially regulated during the process of differentiation induced by phorbol ester, retinoic acid or interferon-gamma. AB - The effects of three inducers of differentiation, phorbol myristate acetate (PMA), retinoic acid (RA) and interferon-gamma (IFN-gamma), on the temporal regulation of vitamin D receptor (VDR) expression in HL-60 cells were analyzed by Northern blotting and immunofluorescence assays. VDR, at the protein level, expressed by 81% of uninduced cells, was reduced to 57% after 48 h of PMA or 96 h of RA treatment, preceded by growth inhibition and cell differentiation, evaluated by CD11b expression. Sorted CD11b positive cells in G0/G1 phase exhibited 53% the VDR content of CD11b negative cells (distributed throughout the cell cycle). PMA also induced an increase in PKC beta and PKC alpha mRNA and protein. Simultaneous exposure to PMA and sphingosine blocked stimulation of CD11b and PKC expression without affecting growth arrest and VDR down regulation. Similar effects were observed during sphingosine treatment. In IFN-gamma differentiated cells, the proportion of cells in G0/G1 phase was unchanged and VDR protein was unaltered as compared to uninduced cells. Control cells in G0/G1 expressed less VDR than cells in S and G2/M phases (74% and 59% respectively). All results suggest that in HL-60 cells, reduction of VDR expression is related to growth inhibition rather than to the differentiation process. PMID- 9744517 TI - Effect of estrogens on IL-1beta promoter activity. AB - It is well documented that steroid hormones modulate cytokine gene expression. In some tissues estrogens are known to suppress cytokine production while in other tissue types, cytokine expression is enhanced by the hormone. This study was conducted to investigate the regulatory mechanisms which underlie the modulation of the interleukin-1beta (IL-1beta) gene at the transcription level. To accomplish this, the macrophage cell line RAW264.7, which appeared insensitive to 17beta-estradiol (E2) treatment, was stably transfected with the human estrogen receptor (ER) and an IL-1beta promoter-CAT reporter construct. E2 markedly enhanced LPS-induced IL-1beta promoter-driven CAT activity in an E2 dose dependent manner. This responsiveness was estrogen specific since no synergism was observed between LPS and the sex steroids testosterone or progesterone while the estrogen analogue 17alpha-estradiol stimulated only at 10 to 100 times the amount required for 17beta-E2. Several antiestrogens, H1285, ICI 182 780, and tamoxifen inhibited the estrogen stimulated enhancement of IL-1beta promoter activity in a dose-dependent manner, indicating that this effect was indeed mediated through the ER in a ligand dependent manner. The estrogenic effect appeared to be indirect and time dependent since the addition of E2 was required hours prior to LPS stimulation; addition of E2 and LPS at the same time resulted in a greatly reduced estrogenic effect. The estrogen metabolites 17-epiestriol and 16-keto-17beta-E2 displayed an estrogenic response virtually indistinguishable from E2. 4-Hydroxyestradiol displayed activity only at 100-fold the concentration of E2 while 2-hydroxyestrone showed no activity at any of the concentrations tested. Overall the results demonstrate that E2 and some metabolites of E2 synergize with LPS to markedly enhance IL-1beta promoter activity through ER mediated processes. PMID- 9744518 TI - Influence of calf serum on glucocorticoid-responses of certain progesterone derivatives. AB - The following in vitro glucocorticoid (GC) parameters of progesterone (P), 1-ene progesterone (deltaP), 11beta-hydroxyprogesterone (HOP), 11beta-1-ene progesterone (deltaHOP) and dexamethasone (Dexa) were assayed in the presence or absence of bovine calf serum (BCS): binding to thymus cytosol, dissociation of the glucocorticoid receptor (GR)-heat shock protein 90 (hsp90) complex (diss.), tyrosine aminotransferase (TAT) induction in hepatocytes and the inhibition of 3H uridine and 35S-methionine uptake by thymocytes. Without BCS, steroids were in most cases active in this general order: Dex > deltaHOP > HOP > deltaP > P. BCS abolished all activities in P and deltaP, but left them unaltered in all other steroids, except diss. in HOP, which diminished intermediately. Binding of P, deltaP, HOP and deltaHOP to GR and CBG paralleled their in vivo activating effects on glycogen deposition. CONCLUSIONS: in this steroid series, BCS, but not CBG, inhibits GC responses of P and deltaP. 11-Beta hydroxylation frees those molecules from the inhibitory effects of BCS. PMID- 9744519 TI - Molecular modelling of steroidogenic cytochromes P450 from families CYP11, CYP17, CYP19 and CYP21 based on the CYP102 crystal structure. AB - The results of homology modelling of mammalian steroidogenic cytochromes P450 (CYP) from families CYP11, CYP17, CYP19 and CYP21 are reported, based on a novel protein sequence alignment with CYP102, a bacterial P450 of known crystal structure. The molecular models generated from the CYP102 crystal structure template are consistent with experimental information from site-directed mutagenesis studies, steroidal substrate specificity and active site inhibitor studies. Interactive docking studies with both substrates and inhibitors of these enzymes indicate key residue interactions with the putative active site regions of each isoform investigated, which point to potential determinants of substrate specificity within these related enzymes. PMID- 9744520 TI - Control of the proliferation of prostate cancer cells by an androgen and two antiandrogens. Cell specific sets of responses. AB - The responses, in terms of cell proliferation and c-myc messenger RNA content, of human prostate cancer cells to androgen-receptor ligands were investigated. Experiments were performed with three types of cells (LNCaP, R2 and MOP) and three compounds (the androgen R 1881 and two anti-androgens: cyproterone acetate, CYPA, and RU 56187). MOP cells were established in the laboratory and the effects of RU 56187 had not been studied in culture. In terms of proliferation, LNCaP was stimulated by the three compounds, R2 was inhibited by R 1881 and RU 56187 but was stimulated by CYPA while MOP was inhibited by the three compounds. In the three types of cells, c-myc messenger RNAs were down regulated by R 1881 and RU 56187 but not by CYPA. The conclusions are: (1) the sets of responses of cell proliferation to three androgen-receptor ligands are cell specific; (2) the control of c-myc messenger RNA by R 1881 and RU 56187 may be related to the inhibition of cell proliferation by these compounds but not to their stimulatory effect on cell proliferation; (3) if prostate tumor cells would respond in vivo to androgens and antiandrogens like in culture, patients with prostate cancer could take benefits of reversible medical castration and sequential prescription of various antiandrogens. PMID- 9744521 TI - Association of vitamin D receptors with the nuclear matrix of human and rat genitourinary tissues. AB - Calcitrol, 1,25 dihydroxyvitamin D3 (1,25-D3) has an important role in the antiproliferative and growth regulatory effects on normal and neoplastic cells (e.g. prostate cancer cells). 1,25-D3 binds to the vitamin D receptor (VDR), a member of the steroid receptor superfamily. Steroids, via intranuclear receptors, have been demonstrated to have high affinity binding to the nuclear matrix, the tissue specific scaffolding of the nucleus that is involved in the organization of DNA, replication and transcription. We hypothesized that the VDR interacts closely with the nuclear matrix in both human and rat tissues. In the studies described here, nuclear matrix proteins (NMP) were extracted from a number of rat and human tissues and immunoblot analysis performed using a rat anti-VDR antibody. The results from these studies reveal that the anti-VDR antibody detects six forms of the VDR in the NMP preparations: human testis demonstrated a protein of 57 and 52 kDa molecular weight compared with 57 and 37 kDa in the rat testis. Human prostate demonstrated proteins of 52 kDa compared to rat ventral (57 and 37 kDa) and dorsal prostate (52 and 26 kDa). Human and rat bladder NMP demonstrated a protein binding at 55 kDa and rat seminal vesicle NMP binding at 48 kDa. This is the first report of VDRs associated with the nuclear matrix. The varying molecular weight proteins reactive with the anti-VDR antibody within these tissues may represent different isoforms, proteolytic cleavage of a larger VDR or post-translational modification. The VDR-NMP interaction may be involved in the tissue specific actions of 1,25-D3 especially growth regulatory and antiproliferative effects. PMID- 9744522 TI - Effects of partial versus pure antiestrogens on ovulation and the pituitary ovarian axis in the rat. AB - The present study was undertaken to compare the effects of the pure antiestrogen ZM 182780 (ZM) and the partial agonist tamoxifen (TAM) on ovulation and peripheral hormone levels in the rat. Adult female rats were treated with ZM (5 mg/kg/d) or TAM (5 mg/kg/d) and sacrificed at varying times during the estrous cycle. ZM and TAM were able to inhibit ovulation to nearly the same extent (60% and 70%, respectively). ZM induced a persistent diestrus whereas TAM led to a mixed picture in the vaginal smear. Both antiestrogens suppressed the preovulatory surge of LH, FSH and progesterone but had no effect on basal FSH values. In contrast to TAM, ZM caused an increase in basal levels of LH. Moreover, basal and preovulatory levels of estradiol and androgens were increased by ZM. By contrast, TAM decreased basal and preovulatory values of LH, estradiol, and androgens. These data suggest that ZM inhibits not only the positive but also negative feedback effect of estrogens and TAM seems to inhibit only the positive feedback. Moreover, it is conceivable that the suppressed preovulatory progesterone surge induced by ZM and TAM could be responsible for the antiovulatory effect. PMID- 9744523 TI - Plasma 25-hydroxyvitamin D concentrations are inversely associated with blood pressure of Dahl salt-sensitive rats. AB - Dietary salt is a contributing factor to the development of hypertension in individuals who are salt-sensitive. The vitamin D endocrine system has been reported to modulate vascular structure and function. Since elderly hypertensive females with low plasma renin activity, typical of salt-sensitivity, had significantly lower 25-hydroxyvitamin D concentrations compared with normotensive elderly and young females, we have used Dahl salt-sensitive and salt-resistant rats fed high (80 g/kg diet) and low (3 g/kg diet) salt diets as models to examine the relationship between salt-sensitivity and 25-hydroxyvitamin D, the precursor of the hormonal form of vitamin D, 1,25-dihydroxyvitamin D. Plasma 25 hydroxyvitamin D concentrations of salt-resistant rats were unaffected by a high salt diet, but plasma 25-hydroxyvitamin D concentrations of salt-sensitive rats were significantly reduced within three weeks to lower than 25%. There was a negative association between plasma 25-hydroxyvitamin D concentrations of salt sensitive rats and the number of days that the rats were fed a high salt diet (r = -0.98, P < 0.02) and a positive association between blood pressure and the number of days that the rats were fed a high salt diet (r = 0.97, P < 0.05). An inverse relationship was found between plasma 25-hydroxyvitamin D concentrations and blood pressure (r = -0.99, P < 0.01). Spontaneously hypertensive rats did not have low plasma 25-hydroxyvitamin D concentrations, suggesting that reduction of plasma 25-hydroxyvitamin D concentration might be specific to salt-induced hypertension. PMID- 9744524 TI - The role of ethnicity in cancer susceptibility gene polymorphisms: the example of CYP1A1. AB - Individual susceptibility to cancer from environmental agents may be influenced by polymorphic metabolic genes such as CYP1A1. The CYP1A1 gene contains four major polymorphisms identified to date. A modern nomenclature system, used with other genes, is presented to clarify the identity of these polymorphisms. The various CYP1A1 alleles exhibit population frequencies that depend on ethnicity. The association of these alleles with cancer at several sites has also been found to depend on racial or ethnic origin of the study population. Statistical considerations, such as the need for large studies when the power to detect a rare polymorphism is low, and ethnic differences in genetic linkage disequilibrium are among possible reasons for ethnic-specific effects on cancer susceptibility related to metabolic gene polymorphisms. New efforts to determine population frequencies of such polymorphisms are essential for future research in this area. PMID- 9744525 TI - The redox-sensitive human antioxidant responsive element induces gene expression under low oxygen conditions. AB - Transient transfection studies of human HepG2 and mouse Hepa hepatocarcinoma cells with a reporter gene construct regulated by a human antioxidant responsive element (ARE) from the NQO1 gene demonstrated that the element is responsive to low oxygen conditions. The antioxidant N-acetyl L-cysteine (NAC) strongly inhibited basal aerobic reporter gene activity in HepG2 cells without obviously affecting the hypoxic induction, as is consistent with ARE sensitivity to oxidative stress in aerobic cultures. Electrophoretic mobility shift (EMS) assays of nuclear extracts of HepG2 and Hepa cells lysed under aerobic or hypoxic conditions or after exposure to the phenolic compound 3-(2)-tert-butyl-4 hydroxyanisole (BHA), showed specific and constitutive protein binding to the ARE under all of these conditions. Taken together, these findings show that the ARE can mediate gene expression in response to low oxygen conditions. Co-ordinately regulated expression of ARE-dependent genes, such as phase II detoxification enzymes, may be an important phenotype of solid tumors containing significant regions of pathophysiological hypoxia. PMID- 9744527 TI - Chemopreventive effects of the aromatase inhibitor vorozole (R 83842) in the methylnitrosourea-induced mammary cancer model. AB - The chemopreventive activity of the highly specific nonsteroidal aromatase inhibitor, vorozole, was examined in the methylnitrosourea (MNU)-induced rat model of mammary carcinogenesis. Various doses of vorozole (0.08-1.25 mg/kg body wt/day) were administered daily (by gavage) to female Sprague-Dawley rats starting at 43 days of age. Seven days later, the rats were given a single i.v. dose of MNU (50 mg/kg body wt). Rats were continually treated with vorozole until the end of the experiment (120 days post-MNU). Vorozole caused a dose dependent inhibition of mammary cancer multiplicity. The highest dose of vorozole (1.25 mg/kg body wt/day) decreased cancer multiplicity by approximately 90%, and simultaneously decreased cancer incidence from 100 to 44%. The next two highest doses of vorozole (0.63 and 0.31 mg/kg body wt/day) inhibited MNU-induced mammary cancer multiplicity by 70-80%. Even the two lowest doses of vorozole (0.16 and 0.08 mg/kg body wt/ day) decreased cancer multiplicity -50%. Serum level determinations were performed on a variety of endpoints at either 4 or 24 h following the last dose of vorozole. Insulin-like growth factor (IGF)-1 levels were slightly, but significantly, increased by vorozole treatment. Vorozole induced striking increases in serum testosterone levels at 4 h at all the dose levels employed. Testosterone levels were significantly elevated over controls at 24 h in rats given the lower doses of vorozole (0.08-0.31 mg/kg body wt/day), but were significantly lower than in rats administered the higher doses of vorozole (0.63 or 1.25 mg/kg body wt/ day). This result presumably reflects the limited half-life of vorozole in rats. In a second series of experiments, the effects of limited duration of dosing with vorozole (2.5 mg/kg body wt/day) or intermittent dosing with vorozole were determined. Treatment of rats with vorozole for limited time periods, from 3 days post-MNU administration until 30 or 60 days post-MNU treatment, resulted in significant delays in the time to appearance of palpable cancers. However, these limited treatments did not greatly affect the overall incidence or multiplicity of mammary cancers when compared with the MNU controls at the end of the study (150 days post-MNU). Finally, the effects of intermittent dosing with vorozole (2.5 mg/kg body wt/day) were examined. Rats were administered cycles of vorozole daily for a period of 3 weeks followed by treatment with the vorozole vehicle for the next 3 weeks (total of four cycles). Although this intermittent treatment did inhibit the appearance of new tumors during each of the periods that vorozole was administered, it did not cause regression of palpable cancers. PMID- 9744526 TI - Differential cleavage of oligonucleotides containing the benzene-derived adduct, 1,N6-benzetheno-dA, by the major human AP endonuclease HAP1 and Escherichia coli exonuclease III and endonuclease IV. AB - We report here that the newly synthesized DNA adduct, 1,N6-benzetheno-dA (pBQ dA), in defined oligonucleotides [Chenna and Singer, Chem. Res. Toxicol., 8, 865 874], is a substrate for the major human AP endonuclease, HAP1, and the Escherichia coli AP endonucleases, exonuclease III and endonuclease IV. The mechanism of cleavage is identical to that reported previously for 3,N4 benzetheno-dC (pBQ-dC) and leads to a phosphodiester bond cleavage 5' to the adduct. There are, however, significant differences in the rate of cleavage of this adduct by these enzymes. The two bacterial AP endonucleases are both much more efficient than the human repair enzyme. In addition, using two random oligodeoxynucleotide sequences containing a single pBQ-dA, exonuclease III and endonuclease IV are similarly active, while HAP1 shows a distinct sequence preference of approximately 10-fold in efficiency of cleavage. The repair of this adduct by the three recombinant enzymes is further confirmed by using both active site mutant HAP1 proteins and by E.coli mutant strains lacking exonuclease III and/ or endonuclease IV. This sequence-dependent repair of pBQ-dA by HAP1 may play an important role in modulating benzene-induced carcinogenesis. PMID- 9744528 TI - Inhibition of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone-induced mouse lung tumor formation by FGN-1 (sulindac sulfone). AB - The sulfone derivative of the non-steroidal anti-inflammatory drug (NSAID), sulindac, has been reported to inhibit mammary and colon tumor formation in rodent models of chemically-induced carcinogenesis. Unlike its parent compound, this metabolite lacks cyclo-oxygenase inhibitory activity. A tumor induction protocol, consisting of NNK administration in the drinking water over several weeks to model chronic human exposure, was used to test whether the sulfone (called FGN-1) could inhibit the formation of primary lung tumors in mice. A total of 150 female, AIN76A-fed, A/J mice received 9 mg of NNK each. Concentrations of FGN-1 that had been previously determined not to affect body weight gain were added to the food at levels of 0, 250, 500 and 750 mg/kg of diet (30 mice/group) starting 2 weeks before NNK administration and continuing for 22 weeks. At that time pleural surface tumors were counted. Tumor incidence decreased significantly from 96 % in the control diet and 93% in the 250 FGN-1 mg/kg diet to 63 and 67% in the 500 and 750 mg FGN-1/kg diet groups, respectively (P < 0.001 by chi-square analysis). Lung tumor multiplicity decreased from 18.1+/ 3 tumors/ mouse (mean+/-SEM, control diet) to 12.3+/-3 (250), 5.3+/-1 (500) and 2.1+/-1 (750) (P < 0.0005 by post hoc ANOVA). In previous studies using this carcinogenesis protocol, the maximum tolerated dose of sulindac inhibited lung tumor multiplicity by no more than 50% with no effect on incidence. This dose dependent reduction in tumorigenesis by a non-toxic dose of FGN-1 indicates a strong chemopreventive activity against experimental induction of lung carcinogenesis. The greater potency of the sulfone over sulindac and its lack of toxic side effects because of its inability to affect cyclo-oxygenase activity suggests that clinical testing in individuals at high risk for lung cancer should be considered. PMID- 9744529 TI - Mechanism of inhibition of benzo[a]pyrene-induced forestomach cancer in mice by dietary curcumin. AB - Curcumin (diferuloylmethane), the major yellow pigment in turmeric, has been shown to inhibit benzo[a]pyrene (BaP)-induced forestomach cancer in mice through mechanism(s) not fully understood. It is well known that while cytochrome P4501A1 (CYP1A1) and epoxide hydrolase (EH) are important in the conversion of BaP to its activated form, (+)-anti-7,8-dihydroxy-9,10-oxy-7,8,9,10-tetrahydrobenzo[a]pyrene [(+)-anti-BaPDE], the detoxification of (+)-anti-BaPDE is accomplished by glutathione (GSH) S-transferases (GST). Therefore, it seems reasonable to postulate that curcumin may exert anti-carcinogenic activity either by inhibiting activation of BaP or (and) by enhancing the detoxification of (+)-anti-BaPDE. Administration p.o. of 2% curcumin in the diet to female A/J mice for 14 days, which has been shown to cause a significant inhibition in BaP-induced forestomach tumorigenesis, resulted in a modest but statistically significant reduction in hepatic ethoxyresorufin O-deethylase (EROD) activity, a reaction preferentially catalyzed by CYP1A1. While EROD activity could not be detected in the forestomach of either control or treated mice, curcumin feeding caused a statistically significant increase (approximately 2.3-fold) in hepatic EH and GST activities. Hepatic and forestomach GSH levels, and forestomach EH and GST activities were not affected by curcumin treatment. Even though the levels of various hepatic GST isoenzymes were significantly increased upon curcumin feeding, maximum induction was noticed for the pi class isoenzyme (mGSTP1-1), which among murine hepatic GSTs is highly efficient in the detoxification of (+)-anti-BaPDE. In conclusion, the results of the present study suggest that curcumin may inhibit BaP-induced forestomach cancer in mice by affecting both activation as well as inactivation pathways of BaP metabolism in the liver. PMID- 9744530 TI - Detection of CYP1A1 protein in human liver and induction by TCDD in precision-cut liver slices incubated in dynamic organ culture. AB - Cytochrome P4501A1 (CYP1A1) has been implicated in the conversion of numerous polycyclic aromatic hydrocarbons into electrophilic species capable of binding covalently to DNA and has therefore been postulated to be involved in the initiation of carcinogenesis. The expression of CYP1A1 protein appears not to be constitutive, but is readily inducible by aryl hydrocarbon (Ah) receptor ligands in a majority of tissues of experimental animals, especially the liver. To date, there is conflicting evidence for the expression or inducibility of CYP1A1 protein in human liver. In this present study, we report the detection of CYP1A1 in all 20 human liver microsomal samples tested by standard western immunoblotting with chemiluminescent detection using a specific monoclonal antibody (mAb 1-12-3) directed against a marine fish (scup) cytochrome P450E. mAb 1-12-3 has been shown previously to specifically recognize CYP1A1 in mammals. This system consistently demonstrated a detection sensitivity as low as 0.01 0.025 pmol CYP1A1 per lane. In the samples where CYP1A1 protein levels were quantitated, CYP1A1 ranged from approximately 0.4 to 5 pmol CYP1A1/mg microsomal protein. Additionally, the inducibility of CYP1A1 protein was demonstrated by incubating precision-cut human liver slices in dynamic organ culture for up to 96 h in the presence of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). The specificity of mAb 1-12-3 was tested using several purified human and rat cytochrome P450s to ensure that the protein being detected was CYP1A1. mAb 1-12-3 did not cross-react with human CYP1A2 or CYP3A4 or rat CYP1B1, but did strongly recognize CYP1A1. However, there was a very weak cross-reactivity of mAb 1-12-3 with human CYP2E1, approximately 75-fold less compared with CYP1A1. In order to confirm CYP1A1 as the immunoreactive protein detected in human liver, microsomal samples were subjected to two-dimensional electrophoresis involving isoelectric focusing followed by SDS-PAGE and immunoblotting. Utilizing mAb 1-12-3, the human liver microsomal samples displayed an immunoblotting profile matching that obtained from a microsomal preparation from a AHH-1 TK+/- cell line expressing solely human CYP1A1 and differing from the profile obtained using a polyclonal antibody directed against CYP2E1 and cells expressing CYP2E1. Furthermore, mAb 1-12-3 recognized only one protein of identical mobility on the two-dimensional blots from human liver microsomes and AHH-1 TK+/- cells expressing CYP1A1, while displaying no reaction to cells expressing only CYP2E1. In conclusion, CYP1A1 appears to be expressed in human liver at low levels and is inducible upon exposure to TCDD. PMID- 9744531 TI - The coffee-specific diterpenes cafestol and kahweol protect against aflatoxin B1 induced genotoxicity through a dual mechanism. AB - The diterpenes cafestol and kahweol (C&K) have been identified in animal models as two potentially chemoprotective agents present in green and roasted coffee beans. It has been postulated that these compounds may act as blocking agents by producing a co-ordinated modulation of multiple enzymes involved in carcinogen detoxification. In this study, we investigated the effects of C&K against the covalent binding of aflatoxin B1 (AFB1) metabolites to DNA. Male Sprague-Dawley rats were treated with increasing amounts of a mixture of C&K in the diet (0-6200 p.p.m.) for 28 and 90 days. A dose-dependent inhibition of AFB1 DNA-binding was observed using S9 and microsomal subcellular fractions from C&K-treated rat liver in an in vitro binding assay. Significant inhibition was detected at 2300 p.p.m. and maximal reduction of DNA adduct formation to nearly 50% of the control value was achieved with 6200 p.p.m. of dietary C&K. Two complementary mechanisms may account for the chemopreventive action of cafestol and kahweol against aflatoxin B1 in rats. A decrease in the expression of the rat activating cytochrome P450s (CYP2C11 and CYP3A2) was observed, as well as a strong induction of the expression of the glutathione-S-transferase (GST) subunit GST Yc2, which is known to detoxify highly the most genotoxic metabolite of AFB1. These data and the previously demonstrated effects of C&K against the development of 7,12 dimethylbenz[a]anthracene (DMBA)-induced carcinogenesis at various tissue sites suggest the potential widespread effect of these coffee components against chemical carcinogenesis. PMID- 9744532 TI - Suppression of mouse skin tumor promotion and induction of apoptosis in HL-60 cells by Alpinia oxyphylla Miquel (Zingiberaceae). AB - There have been considerable efforts to search for naturally occurring substances for the intervention of carcinogenesis. Many components from dietary or medicinal plants have been identified that possess substantial chemopreventive properties. An example is curcumin (Curcuma longa Linn., Zingiberaceae), which has been shown to inhibit tumor promotion in experimental carcinogenesis. Alpinia oxyphylla Miquel, another plant of the ginger family used in oriental herbal medicine, contains diarylheptanoids whose structures are analogous to that of curcumin. In the present study, we have tested A.oxyphylla for its ability to suppress tumor promotion. Thus, topical application of the methanolic extract of dried fruits of A.oxyphylla significantly ameliorated 12-O-tetradecanoylphorbol-13-acetate (TPA) induced skin tumor promotion as well as ear edema in female ICR mice. In another study, treatment of HL-60 cells with the methanolic extract of A.oxyphylla significantly reduced the viability of the cells and also inhibited DNA synthesis. Microscopic examination of the treated cells showed characteristic morphology of apoptosis. Furthermore, cells treated with the extract of A.oxyphylla exhibited internucleosomal DNA fragmentation in time- and concentration-dependent manners. TPA-stimulated generation of superoxide anion in differentiated HL-60 cells was also blunted by A.oxyphylla. Taken together, these findings suggest that A.oxyphylla possesses potential chemopreventive and antitumorigenic activities. PMID- 9744533 TI - Alcohol-related cancers and aldehyde dehydrogenase-2 in Japanese alcoholics. AB - Aldehyde dehydrogenase-2 (ALDH2) eliminates most of the acetaldehyde produced during alcohol metabolism. In some drinkers, a mutant ALDH2 allele contributes to diminished activity of the enzyme, dramatically increasing the risk for esophageal cancer. This study was designed to evaluate the ALDH2 gene polymorphism as a predictor of the development of cancers prevalent in Japanese alcoholics. We performed ALDH2 genotyping on lymphocyte DNA samples from Japanese alcoholic men (487 cancer-free; 237 with cancer, including 34 oropharyngolaryngeal, 87 esophageal, 58 stomach, 46 colon, 18 liver, 7 lung, 9 other sites, and 19 multiple primary cancers in two or three organs). The frequencies of the mutant ALDH2*2 allele were significantly higher in alcoholics with oropharyngolaryngeal (52.9%), esophageal (52.9%), stomach (22.4%), colon (21.7%) and esophageal cancer concomitant with oropharyngolaryngeal and/or stomach cancer (78.6%), than in cancer-free alcoholics (9.0%). After adjustment for age, daily alcohol consumption and amount of cigarette smoking, significantly increased risks (odds ratios) in the presence of the ALDH2 *2 allele were found for oropharyngolaryngeal (11.14), esophageal (12.50), stomach (3.49), colon (3.35), lung (8.20) and esophageal cancer concomitant with oropharyngolaryngeal and/or stomach cancer (54.20) but not for liver or other cancers. These results suggest a general role of acetaldehyde, a recognized animal carcinogen, in the development of human cancers. PMID- 9744534 TI - Polycyclic aromatic hydrocarbon-DNA adducts in human placenta and modulation by CYP1A1 induction and genotype. AB - This study investigated the relationship in human placenta between polycyclic aromatic hydrocabon (PAH)-DNA adduct levels and two biomarkers of cytochrome P4501A1 (CYP1A1): gene induction evidenced by CYP1A1 mRNA, and a genetic polymorphism, the CYP1A1 MspI RFLP. CYP1A1 codes for an inducible enzyme system that catalyzes the bioactivation of PAHs. Prior research found a high correlation in human lung tissue between CYP1A1 activity and DNA damage from PAHs. The CYP1A1 Mspi RFLP has been linked in some studies to risk of lung cancer. The relationships in human placenta between DNA damage, CYP1A1 activity and genotype have not been well characterized and may be relevant to risks from transplacental PAH exposure. The study cohort consisted of 70 newborns from Krakow, Poland, a city with elevated air pollution, and 90 newborns from nearby Limanowa, an area with lower air pollution but greater indoor coal use. Contrary to results seen previously in lung tissue, CYP1A1 mRNA was not significantly correlated with PAH DNA adduct levels in the placenta. Smoking (self-reported maternal and infant plasma cotinine) was significantly associated with CYP1A1 mRNA levels (P < 0.01), but not with PAH-DNA adduct levels. Placental PAH-DNA adduct levels were significantly higher in infants with the CYP1A1 MspI restriction site compared with infants without the restriction site (P < 0.01), implicating a genetic factor in inter-individual variation in DNA damage in human placenta. Further studies are needed to determine the relevance of this finding to risk of transplacental carcinogenesis. PMID- 9744535 TI - Inhibitors of lipoxygenase: a new class of cancer chemopreventive agents. AB - 5-Lipoxygenase is a key enzyme in the metabolism of arachidonic acid to leukotrienes. The preventive efficacy of 5-lipoxygenase inhibitors against lung tumorigenesis was determined in A/J mice given the tobacco-specific carcinogen 4 (methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) in drinking water from week 0 to week +7. Groups of 25 mice were fed either: acetylsalicylic acid (ASA), a cyclooxygenase inhibitor; A-79175, a 5-lipoxygenase inhibitor; MK-886, an inhibitor of the 5-lipoxygenase activating-protein; a combination of ASA and A 79175 from weeks -2 to +23. ASA, A-79175 and MK-886 reduced lung tumor multiplicity by 44, 75 and 52% respectively. Furthermore, A-79175 reduced tumor incidence by 20%. Administration of A-79175 and MK-886 decreased the mean tumor volume by 64 and 44% respectively. Lung tumor multiplicity was directly proportional to tumor volume. The combination of ASA and A-79715 was the most effective preventive intervention and reduced lung tumor multiplicity by 87% and lung tumor incidence by 24%, demonstrating that inhibition of both 5-lipoxygenase and cyclooxygenase is more effective than inhibition of either pathway alone. NNK treatment increased plasma prostaglandin E2 levels from 49 to 260 pg/ml and plasma LTB4 levels from 29 to 71 pg/ml. Incubation of 82-132 and LM2 murine lung tumor cells with MK-886 and A-79715 decreased cell proliferation in a concentration-dependent manner. Soybean lipoxygenases with or without murine lung microsomal proteins metabolized NNK by alpha-carbon hydroxylation (9.5% of the metabolites) and N-oxidation (3.9%). Activation of NNK by alpha-carbon hydroxylation was inhibited by addition of arachidonic acid and A-79715. Possible mechanisms of action of 5-lipoxygenase inhibitors include inhibition of tumor growth and lipoxygenase-mediated activation of NNK. These studies suggest that inhibitors of 5-lipoxygenase may have benefits as preventive agents of lung tumorigenesis. PMID- 9744536 TI - Effects of Cr(VI) on the expression of the oxidative stress genes in human lung cells. AB - Intracellular metabolism of chromium(VI) [Cr(VI)] may lead to oxidative stress and this may account for the ability of Cr(VI) to act as a complete carcinogen. Therefore, we examined the effects of Cr(VI) treatment on the expression of oxidative stress genes in normal human lung LL 24 cells and human lung adenocarcinoma A549 cells. RT-PCR and northern blot analyses were used to determine the steady-state mRNA levels of catalase, glutathione S-transferase, glutathione reductase, Cu/Zn- and Mn-superoxide dismutases, glutathione peroxidase, NAD(P)H:quinone oxidoreductase, heme oxygenase and interleukin 8 in control cells and cells treated with 5-200 microM of Cr(VI). We found that only expression of the heme oxygenase gene is strongly elevated under the treatment with Cr(VI), and only in normal human lung LL 24 cells. Our data showed that even in the absence of Cr(VI) treatment, the level of heme oxygenase gene expression is much higher in A549 cells than in LL 24 cells. As glutathione is believed to play a protective role in cells against different forms of oxidative stress, we studied the correlation between intracellular glutathione levels and the inducibility of the heme oxygenase gene after treatment of cells with Cr(VI). Our results demonstrate that glutathione levels are increased by 35 % of control values in LL 24 cells treated with Cr(VI). The data obtained indicate that heme oxygenase, known to be a stress-inducible gene, may be involved in cellular pathways critical to the carcinogenic activity of Cr(VI) in normal human lung cells. Intracellular glutathione levels and reactive oxygen species do not appear to be primarily responsible for the stress response, induced by Cr(VI) in the studied human cells. PMID- 9744537 TI - Co-operation between follicular ornithine decarboxylase and v-Ha-ras induces spontaneous papillomas and malignant conversion in transgenic skin. AB - Ornithine decarboxylase (ODC) is aberrantly regulated in tumor cells and results in high basal levels of ODC and polyamines in many epithelial tumors. To determine if elevated ODC/polyamine levels can co-operate with a mutant Ha-ras gene in mouse skin tumorigenesis, double transgenic mice were generated by breeding K6/ODC transgenic mice with TG.AC v-Ha-ras transgenic mice. A K6 keratin promoter drives the ODC transgene in K6/ ODC transgenic mice, which results in elevated ODC/ polyamine levels directed to the outer root sheath cells of hair follicles. TG.AC transgenic mice carry a v-Ha-ras transgene while still retaining two normal c-Ha-ras alleles. Transgenic mice that possess only the K6/ODC or the v-Ha-ras transgene did not develop tumors unless treated with either a carcinogen or a tumor promoter, respectively. However, a high percentage of double transgenic mice possessing both the K6/ODC and v-Ha-ras transgenes developed spontaneous tumors. All tumors were well-differentiated keratoacanthomas, some of which progressed to carcinomas within 2 months. The development and the maintenance of these ODC/ras tumors was ODC-dependent since alpha difluoromethylornithine (DFMO), a specific ODC inhibitor, prevented the formation and caused the regression of these tumors. These findings indicate that ODC overexpression and an activated Ha-ras are sufficient to produce a high rate of malignant transformation in an animal model. The ODC/ras double transgenic mouse provides a simple in vivo model without the use of chemical carcinogens or tumor promoters in which to test downstream effectors that play a key role in mediating the development of epithelial tumors resulting from the cooperation between ODC and v-Ha-ras. PMID- 9744538 TI - The effect of short-term fasting, phenobarbital and refeeding on apoptotic loss, cell replication and gene expression in rat liver during the promotion stage. AB - Previous work from this laboratory has reported on the effects of two sequential 5 day periods of fasting and subsequent refeeding on tumor promotion in multistage hepatocarcinogenesis in the rat (Carcinogenesis, 18, 159-166, 1997). In the present extension of the earlier study, the sequential fasting-refeeding regimen was begun at later time points (28 and 54 days post-initiation) than the first study. This was done to determine whether larger-sized altered hepatic foci (AHF) exhibited a depletion similar to that of the relatively small AHF in the published experiment and to study concomitant molecular changes during the fasting periods. Groups of animals were fasted in the presence and absence of 0.05% phenobarbital (PB) in the drinking water. During the fasting periods, both body and liver weights decreased dramatically, less in the fast begun at 54 days. This change was accompanied by a significant decrease in the bromodeoxyuridine (BrdU) labeling indices of hepatocytes within AHF. Apoptotic bodies increased dramatically in the non-focal (surrounding the AHF) hepatocytes during the fasting periods. These parameters were slightly lower in hepatocytes of rats administered PB during the fasting periods, most notably during the 54-66 day period. With the nick end-labeling method, the proportion of hepatocytes undergoing apoptosis was significantly higher in cells within AHF at the end of each of the fasting periods in all but one group. Concomitantly, the number of AHF and percentage of liver volume occupied by AHF decreased dramatically during the fasting periods. Refeeding caused a marked increase in BrdU labeling in hepatocytes within and surrounding AHF during the first week or two, most notably in animals not receiving PB during the fasting period. Both the number and volume percentage of liver AHF returned to control values within approximately 2 weeks of the refeeding regimen. Assays of nuclear DNA fragmentation with samples of whole liver indicated that a 'laddering' effect was most noticeable in livers of animals subjected to the fasting-refeeding regimen when phenobarbital was not present during the fasting period. Studies of the levels of mRNA of several genes in the total liver revealed that the expression of c-myc increased 3- to 9-fold during the fasting periods but rapidly returned to normal levels after refeeding. Levels of albumin and insulin-like growth factor I mRNAs decreased significantly during the fasting period, but rapidly reappeared on refeeding. These results indicate that the extensive loss of AHF during the short-term fasting periods occurs even when the number and volume of AHF are 10- to 50-fold greater at the beginning of the fast than the values published previously. Both the decrease in insulin growth factor I and the elevation of c-myc expression during the fasting period may indicate the role of these genes in the transcriptional regulation of hepatocyte apoptosis in both normal and preneoplastic hepatocytes in the rat. PMID- 9744539 TI - Differences in kinetics of induction and reversibility of TCDD-induced changes in cell proliferation and CYP1A1 expression in female Sprague-Dawley rat liver. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a potent tumor promoter in two stage initiation-promotion models and induces cell proliferation and development of enzyme-altered hepatic foci. It is believed that increased cell proliferation is a necessary step in carcinogenesis. Therefore, the analysis of the effect of TCDD on cell proliferation in rat liver may aid in the understanding of the mechanism of hepatocarcinogenesis induced by TCDD. The aim of this study was to investigate the time course and reversibility of cell proliferation in non initiated and diethylnitrosamine-initiated female rats exposed biweekly to a daily averaged dose of 125 ng TCDD/kg/day for up to 60 weeks. In addition we evaluated the suitability of different dose metrics for the evaluation of TCDD induced changes in cell proliferation and CYP1A1 enzyme induction. Cell proliferation was measured as the incorporation of 5-bromo-2'-deoxyuridine (BrdU) into hepatocytes undergoing replicative DNA synthesis. Mean BrdU labeling indices in TCDD-treated animals were not increased over controls after 14 weeks exposure, but were increased 8- and 2-fold after 30 and 60 weeks' treatment respectively, despite similar liver levels of TCDD at all these times (23-30 p.p.b.). In comparison, CYP1A1 activity, as measured by ethoxyresorufin deethylase activity, was significantly induced at all times points analyzed. Sixteen weeks following cessation of TCDD treatment, labeling indices were still significantly elevated over controls, but after 30 weeks of withdrawal, labeling indices were no different from controls, indicating that TCDD-induced changes in cell proliferation were reversible. Dosimetric analysis indicated that rat liver tissue burden was suitable for prediction of CYP1A1 expression but not cell proliferation and that the area under the curve was unsuitable for prediction of both TCDD-induced changes in CYP1A1 expression and cell proliferation. PMID- 9744540 TI - Expression of CYP1B1 but not CYP1A1 by primary cultured human mammary stromal fibroblasts constitutively and in response to dioxin exposure: role of the Ah receptor. AB - The expression of CYP1B1 in human mammary fibroblasts (HMFs) was characterized as a potential modulator of their individual function as well as effects on adjacent mammary epithelia. We have used these characteristics to explore the diversity of fibroblast cells isolated from reduction mammoplasty patients and from different breast locations in breast cancer patients (tumors, peripheral to tumor and skin). These parameters have also been used to examine differences between two donors. The results have shown that while none of these HMFs expressed a detectable CYP1A1 protein basally or in response to TCDD, they all expressed CYP1B1 constitutively at similar levels (0.5-0.9 pmol/mg microsomal proteins) and they were induced by TCDD (up to 5-fold) consistent with mediation by the Ah receptor (AhR). DMBA metabolism by HMFs exhibited high proportions of 5,6-, 10,11 and 3,4-dihydrodiols, a profile that is typical of human CYP1B1 regioselectivity. RT-PCR followed by Southern blot analyses demonstrated that CYP1B1 mRNA expression in HMFs parallels levels of respective microsomal proteins. The AhR is expressed in these HMFs as two cytosolic forms (approximately 106 and 104 kDa) and a substantial proportion of the 104 kDa form was localized to the nucleus even prior to TCDD treatment. In all HMFs isolated directly from collagenase digested breast tissues the AhR is expressed at levels 10-fold lower than in breast epithelial cells. However, HMFs that were isolated after serial passaging of mammary epithelial cultures had shown much higher levels of the AhR expression and more dramatic TCDD-induced down-regulation (>80% in 24 h) associated with more efficient nuclear translocation. These differences suggested the presence of two functionally distinct subtypes of HMFs: interstitial stromal fibroblasts that are readily released by collagenase digestion of breast tissues, and lobular stromal fibroblasts which are more tightly associated with the breast epithelia. PMID- 9744541 TI - Studies of iron deposits, inducible nitric oxide synthase and nitrotyrosine in a rat model for esophageal adenocarcinoma. AB - A rat model was developed recently in our laboratory to study the pathogenesis of Barrett's esophagus (BE) and its progression to esophageal adenocarcinoma (EAC). Eight-week-old male Sprague-Dawley rats underwent esophagoduodenal anastomosis (EDA) to produce gastric and duodenal reflux in their distal esophagi. The rats were given iron dextran (50 mg of Fe/kg, i.p.) starting 2 weeks after surgery and this was continued once a month. BE was observed as early as week 3 and the incidence of BE and EAC increased with time: 58 and 17% at week 23; 91 and 73% at week 31. There was a progression in epithelial cell proliferation and inflammation from mild to severe in the distal one-third of the esophagus. Iron deposition in the esophagus also increased with time. Iron deposits in the stromal tissue adjacent to the epithelium in the distal one-third of the esophagus were associated with areas of severe inflammation. Immunohistochemical analysis showed positive inducible nitric oxide synthase (iNOS) expression in the stromal macrophages directly beneath the epithelium in the distal one-third of the esophagus in 36, 83 and 100% of the rats at weeks 17, 23 and 31, respectively. A significant increasing linear trend (P=0.001) was seen in nitrotyrosine immunostaining (number of positive cells/high power field) in the distal esophagus. Strong positive nitrotyrosine staining was seen in the macrophages and weaker positive staining was seen in the adjacent epithelium starting at week 17. Furthermore, iron supplemented rats killed at week 31 had significantly higher (P < 0.05) levels of inflammation, cell proliferation, iNOS and nitrotyrosine as well as more tumors in their distal esophagi than did rats that received no iron supplement. These results suggest that iron supplementation enhanced inflammation and the production of reactive oxygen and nitrogen species in the esophageal epithelium. These processes could contribute to the formation of BE and its progression to EAC. PMID- 9744542 TI - Identification of differentially expressed genes in aflatoxin B1-treated cultured primary rat hepatocytes and Fischer 344 rats. AB - Aflatoxin B1 (AFB1), a mutagen and hepatocarcinogen in rats and humans, is a contaminant of the human food supply, particularly in parts of Africa and Asia. AFB1-induced changes in gene expression may play a part in the development of the toxic, immunosuppressive and carcinogenic properties of this fungal metabolite. An understanding of the-role of AFB1 in modulating gene regulation should provide insight regarding mechanisms of AFB1-induced carcinogenesis. We used three PCR based subtractive techniques to identify AFB1-responsive genes in cultured primary rat hepatocyte RNA: differential display PCR (DD-PCR), representational difference analysis (RDA) and suppression subtractive hybridization (SSH). Each of the three techniques identified AFB1-responsive genes, although no individual cDNA was isolated by more than one technique. Nine cDNAs isolated using DD-PCR, RDA or SSH were found to represent eight genes that are differentially expressed as a result of AFB1 exposure. Genes whose mRNA levels were increased in cultured primary rat hepatocytes after AFB1 treatment were corticosteroid binding globulin (CBG), cytochrome P450 4F1 (CYP4F1), alpha-2 microglobulin, C4b-binding protein (C4BP), serum amyloid A-2 and glutathione S-transferase Yb2 (GST). Transferrin and a small CYP3A-like cDNA had reduced mRNA levels after AFB1 exposure. Full length CYP3A mRNA levels were increased. When liver RNA from AFB1-treated male F344 rats was evaluated for transferrin, CBG, GST, CYP3A and CYP4F1 expression, a decrease in transferrin mRNA and an increase in CBG, GST, CYP3A and CYP4F1 mRNA levels was also seen. Analysis of the potential function of these genes in maintaining cellular homeostasis suggests that their differential expression could contribute to the toxicity associated with AFB1 exposure. PMID- 9744543 TI - DNA modifications by the mutagen glyoxal: adduction to G and C, deamination of C and GC and GA cross-linking. AB - The mutagen glyoxal was reacted with DNA or deoxynucleosides under physiological conditions (pH 7.4, 37 degrees C) in vitro and the products were analyzed by HPLC coupled with a photodiode array UV detector. The efficient formation of a cyclic glyoxal-dG adduct (dG+) was observed in DNA, as well as with dG. The monomeric dG + was gradually decomposed to dG at pH 7.4 and 37 degrees C (t1/2 14.8 h). However, the dG+ formed in single- and double-stranded DNA was rather stable under physiological conditions and the half-lives were 19 and 40 times longer respectively than that of the monomer (t1/2 285 and 595 h respectively). By reaction of glyoxal with deoxycytidine (dC), the deamination products deoxyuridine and 5-hydroxyacetyl-dC (dC+) were formed. Under the same conditions, 5-methyl-dC was deaminated to dT at a higher rate. Deoxyuridine was also formed in DNA by glyoxal treatment. When glyoxal was reacted with various combinations of deoxynucleosides for a prolonged period, dG-glyoxal-dC (GgC), dG-glyoxal-dA (GgA), dG-glyoxal-dG (GgG) and dC-glyoxal-dC (CgC) cross-links were detected, although structures were not assigned unequivocally. Among these, the former two, the GC and GA cross-links, were detected in glyoxal-treated DNA. The yields of these products in DNA were in the following order; dG+ > dU > GgA > GgC > dC+. These DNA modifications may be relevant to glyoxal-induced mutations at GC pairs. PMID- 9744544 TI - Induction of reactive oxygen species without 8-hydroxydeoxyguanosine formation in DNA of initiated mouse keratinocytes treated with 12-O-tetradecanoylphorbol-13 acetate. AB - Evidence for the involvement of oxidative stress in 12-O-tetradecanoylphorbol-13 acetate (TPA)-mediated tumor promotion has focused on non-initiated immune cells, tumor cell lines and non-initiated epidermis treated in vivo. This paper reports the effects of TPA on 8-hydroxydeoxyguanosine (8OHdG) formation and the generation of reactive oxygen species (ROS) in cloned initiated mouse epidermal keratinocytes in order to determine whether TPA can directly damage DNA through ROS production within the keratinocytes. Using high performance liquid chromatography with electrochemical detection (HPLC-EC), TPA did not induce 8OHdG formation in DNA of initiated keratinocytes treated under a variety of conditions. The reliability of the HPLC-EC system is demonstrated by (i) the linearity of the 8OHdG standard curve; (ii) the consistency of 8OHdG measurements in calf thymus and cellular DNA; and (iii) the dose-dependent increase in 8OHdG in DNA of initiated keratinocytes treated with UVC in the presence and absence of H2O2. Though not DNA-damaging, TPA induced a 65% increase in ROS (P < 0.05) as detected by luminol-dependent chemiluminescence. These results support a mechanism for the role of oxidative stress in tumor promotion that does not involve direct DNA damage to the keratinocyte target cell. The relationship between ROS, signal transduction and tumor promotion is discussed in light of the above results which is consistent with the role of TPA-induced ROS as second messengers in signal transduction. PMID- 9744545 TI - Presence of a threshold for promoting effects of phenobarbital on diethylnitrosamine-induced hepatic foci in the rat. AB - The dose dependence of the hepatopromoting effects of phenobarbital (PB) was investigated in a rat liver medium-term bioassay (Ito test) to elucidate a practical threshold level. F344 rats were given a single i.p. injection of diethylnitrosamine (200 mg/kg body wt) and subjected to two-thirds partial hepatectomy at week 3. Commencing 2 weeks from the start, PB at doses of 0, 1, 2, 4, 7.5, 15 or 500 p.p.m. in experiment 1 and 0, 0.01, 0.1 or 0.5 p.p.m. in experiment 2 were fed to the rats for 6 weeks. Experiment 3 was conducted to confirm previous data using the same medium-term bioassay, with PB at doses of 0, 1, 2, 4, 7.5, 15, 30, 60, 125, 250 or 500 p.p.m. fed to the rats. All surviving animals were killed at week 8 in these experiments and their livers were immunohistochemically examined for expression of glutathione S-transferase placental form (GST-P). Quantitative values for GST-P-positive foci in the liver were increased dose dependently in rats given 60-500 p.p.m. PB. However, those for doses in the range 1-7.5 p.p.m. demonstrated a decrease as compared with the control group (0 p.p.m.), with significant differences observed for 1 and 2 p.p.m.. The results for 15-30 and 0.01-0.5 p.p.m. were comparable with the control values. Examination of transforming growth factor-alpha (TGF-alpha) positive foci also produced similar results to those for GST-P in experiment 1. Immunohistochemical staining of TGF-alpha and GST-P using serial liver sections demonstrated that the TGF-alpha-positive foci comprised a sub-population of the GST-P-positive lesions, being approximately 1/8-1/10th as common in livers of animals treated with PB. TGF-alpha-positive foci were almost always negative on immunostaining for TGF-beta. Western blotting for proteins CYP2B1, 2C6 and 3A2 revealed a good correlation between changes in GST-P-positive foci and CYP3A2 protein expression. The finding of inhibition effects at low doses of PB confirms the presence of a threshold level for promoting effects by PB on liver carcinogenesis in rats. PMID- 9744547 TI - Hypomethylation of the rat glutathione S-transferase pi (GSTP) promoter region isolated from methyl-deficient livers and GSTP-positive liver neoplasms. AB - DNA methylation at the 5-position on the cytosine ring in CpG dinucleotides (CpG sites) appears to play an important role in regulating gene expression. In general, there is an inverse relationship between promoter CpG site methylation and the potential for transcription. Thus, changes in DNA methylation density may lead to altered levels of proteins such as glutathione S-transferase pi (GSTP), which is frequently used as a marker to detect hepatocellular foci and neoplasms in the rat. In the present study, the level of CpG methylation in the rat GSTP promoter region was determined in bisulfite-treated DNA isolated from control (untreated) rat livers, chemically induced, GSTP-positive rat liver neoplasms, and methyl-deficient rat livers that contained numerous GSTP-positive foci after administration of a defined diet deficient in folate and choline and low in methionine (0.18%). Eight cytosines between -235 and + 140 in the GSTP promoter region were methylated in a site-specific manner in GSTP-negative control liver, whereas these same sites were hypomethylated in all four chemically-induced, GSTP positive neoplasms. Similarly, all CpG sites were unmethylated in methyl deficient liver DNA within 3 weeks of the rats receiving the methyl-deficient diet, and they remained unmethylated throughout the 36-week treatment period. Five of the eight CpG sites are located within consensus sequences for the DNA binding proteins Spl and E2F. This indicates at least one possible mechanism that could potentially lead to transcriptional activation of GSTP in hepatocellular foci and neoplasms during rat hepatocarcinogenesis. These findings suggest that methylation of critical cytosines within the promoter region rather than all CpG associated cytosines may be a determining factor in regulation of GSTP expression. PMID- 9744546 TI - HPV-16 E7 protein bypasses keratinocyte growth inhibition by serum and calcium. AB - The E6 and E7 genes of HPV-16 or HPV-18 both are necessary for effective immortalization of primary human genital keratinocytes. To analyse the individual role of E6 and E7 genes in dysregulating cell growth, we cloned the HPV-16 E6, E7 and E6/E7 genes into retroviruses. Primary human keratinocytes (PHK) were then infected with these retroviruses and selected in differentiation-inducing medium (high calcium and serum). The E6/E7 retroviruses were the most effective at inducing differentiation-resistant colonies. Intermediate numbers of colonies were induced by E6 and low numbers by E7. Interestingly, only cultures infected with E7 and E6/E7 retroviruses showed a significant proportion of cells progressing into the S phase, consistent with our earlier studies showing that E7 is required for the efficient immortalization of genital keratinocytes. Accompanying this entry into S phase, the E7 or E6/E7 transduced cells expressed high levels of cyclins A, B and E, but lower levels of cyclin D. In addition, cdc 2, cdk-2 and cdk-4 were also increased. No significant differences were detected in the expression of c-myc and c-fos between the vector and any of the transduced cells. Keratinocytes infected with the E7 retrovirus exhibited decreased levels of Rb protein and increased levels of p53, whereas cells infected with E6 expressing retroviruses displayed normal levels of Rb protein and decreased levels of p53. Finally, E7 induced a three-fold increase in bcl-2 expression. Our results indicate that the HPV-16 E7 gene alone is sufficient to bypass keratinoctye growth arrest induced by serum and calcium exposure and that the discordant expression of several cell regulatory proteins accompanies this unregulated proliferation. PMID- 9744548 TI - Induction of hepatic CYP1A in channel catfish increases binding of 2 aminoanthracene to DNA in vitro and in vivo. AB - Data are presented from in vitro and in vivo studies that indicate cytochrome P4501A (CYP1A) in channel catfish (Ictalurus punctatus) hepatic tissue activates 2-amino-anthracene (AA) to a reactive metabolite that binds to DNA. Channel catfish were injected i.p. with vehicle or 10 mg/kg beta-naphthoflavone (betaNF) on two consecutive days. Two days after the final injection of vehicle or betaNF, vehicle or [3H]AA was injected i.p. at 10 mg/kg, creating four different treatments: vehicle only, betaNF only, [3H]AA only, and betaNF/[3H]AA. Hepatic tissue was examined for CYP1A-associated ethoxyresorufin-O-de-ethylase (EROD) activity, and for DNA adducts at 1, 2, 4 and 7 days following administration of vehicle or [3H]AA. Hepatic EROD activity in betaNF-treated fish was 17-fold higher at day 0 and remained significantly greater than untreated animals for the 7-day experiment. Hepatic DNA adducts, as measured by tritium-associated DNA, ranged from 4.8 to 8.6 pmol/mg DNA in vehicle-pretreated fish injected with [3H]AA, but ranged from 12.6 to 22.7 pmol/mg DNA in betaNF-pretreated fish injected with [3H]AA. Thus, pretreatment with betaNF significantly increased binding of [3H]AA to hepatic DNA in vivo at all four times. Analysis by 32P-post labeling and thin layer chromatography of hepatic DNA from channel catfish treated with AA revealed two major and several minor spots, which are indicative of DNA adduct formation. Hepatic microsomes from betaNF-pretreated fish were more effective at catalysing the binding of [3H]AA to DNA in vitro than were microsomes from non-treated fish. In addition, binding was decreased by the CYP1A inhibitor 3,3',4,4'-tetrachlorobiphenyl. Collectively, these data demonstrate that CYP1A is involved in the activation of AA in channel catfish. PMID- 9744549 TI - Effect of promoter and intron 2 polymorphisms on murine lung K-ras gene expression. AB - Differences in tumor formation among inbred mouse strains with high (A/J) and low (C3H) susceptibility for lung cancer have been linked to a repetitive element within the second intron of the K-ras gene. The purpose of this investigation was to determine whether differences within the K-ras gene promoter region or the intron 2 polymorphism affect K-ras gene expression in lung tumors and target alveolar type II cells isolated from A/J and C3H mice. Ribonuclease protection assays were performed using RNA isolated from 4-(methylnitrosamino)-1-(3-pyridyl) 1-butanone (NNK)-induced lung tumors from each mouse strain and alveolar type II cells isolated from A/J and C3H mice. An 838 bp fragment of the murine K-ras gene promoter region was amplified by PCR, cloned and sequenced from both mouse strains. Promoter regions from both mouse strains were inserted into a luciferase reporter gene vector, with and without the second intron polymorphism, and transfected into sensitive, intermediate and resistant lung tumor cell lines. No significant differences in K-ras gene promoter activity was found between the two strains using these specific reporter gene constructs. Consistent with these results, levels of K-ras expression did not differ between alveolar type II cells, whole lung or tumors induced by NNK in A/J or C3H mice. Moreover, in lung tumor cell lines derived from mice with differing susceptibility for lung cancer, K-ras expression did not correlate with the growth rate of tumors induced in nude mice from these cell lines. These results indicate that factors involved in modulating the rapid clonal expansion of the mutated K-ras allele from A/J mice are not directly linked to expression of this gene. Other genetic changes or losses in conjunction with hypothesized modifier loci, such as the Par1 locus, must play a significant role in establishing the phenotypic strain differences for lung tumor formation. PMID- 9744550 TI - Isoflavone genistein inhibits the initiation and promotion of two-stage skin carcinogenesis in mice. AB - Isoflavone genistein is a specific inhibitor of protein tyrosine kinase (PTK) and has been shown to have a variety of anticancer activities in cultured cells and animal models. We report here that genistein significantly inhibits 7,12 dimethylbenz[a]anthracene (DMBA)-initiated and 12-O-tetradecanoyl phorbol-13 acetate (TPA)-promoted skin tumorigenesis in a two-stage carcinogenesis model. In an initiation study, 10 micromol genistein was applied daily to female SENCAR mouse skin for 1 week, followed by initiation with 10 nmol DMBA. Mice were then treated with twice weekly 4 microg TPA. Genistein was shown to reduce tumor incidence and multiplicity in DMBA-initiated skin tumors by approximately 20 (P < 0.05) and 50% (P < 0.01), respectively. Two promotion studies were conducted using CD-1 and SENCAR mice. In experiment 1, CD-1 mice were initiated with 100 nmol DMBA and followed by a twice weekly regimen of 1 and 5 micromol genistein/4 microg TPA. In experiment 2, SENCAR mice were initiated with 10 nmol DMBA and followed by a regimen of 5, 10 and 20 micromol genistein/2 microg TPA. Both studies consistently showed that genistein substantially inhibited TPA-promoted skin tumorigenesis by reducing the tumor multiplicity by approximately 60 and 75%, respectively (P < 0.01). However, the tumor incidence appeared to be less affected. Mechanistic studies showed that genistein inhibited DMBA-induced bulky DNA adduct formation and substantially suppressed TPA-stimulated H2O2 and inflammatory responses in mouse skin by >60% (P < 0.01). In contrast, genistein only exhibited a moderate inhibition of TPA-induced ornithine decarboxylase activity (P > 0.05). Our results suggest that genistein exerts its anti initiational and anti-promotional effects on skin carcinogenesis probably through blockage of DNA adduct formation and inhibition of oxidative and inflammatory events in vivo. PMID- 9744551 TI - Inhibition of cigarette smoke-related lipophilic DNA adducts in rat tissues by dietary oltipraz. AB - The present study investigated the effects of dietary oltipraz on cigarette smoke related lipophilic DNA adduct formation. Female Sprague-Dawley rats were exposed daily to sidestream cigarette smoke in a whole-body exposure chamber 6 h/day for 4 consecutive weeks. One group of rats was maintained on control diet while another group received the same diet supplemented with either a low (167 p.p.m.) or high (500 p.p.m.) dose of oltipraz, starting 1 week prior to initiation of smoke exposure until the end of the experiment. Analysis of lipophilic DNA adducts by the nuclease P1-mediated 32P-post-labeling showed up to five smoke related adducts. Adduct no. 5 predominated in both the lung and the heart while adduct nos 3 and 2 predominated in the trachea and bladder, respectively. Quantitative analysis revealed that the total adduct level was the highest in lungs (270+/-68 adducts/10(10) nucleotides), followed by trachea (196+/-48 adducts/10(10) nucleotides), heart (141+/-22 adducts/10(10) nucleotides) and bladder (85+/-16 adducts/10(10) nucleotides). High dose oltipraz treatment reduced the adduct levels in lungs and bladder by >60%, while the reduction in lungs in the low-dose group was approximately 35%. In trachea, the effect of low and high dietary oltipraz on smoke DNA adduction was equivocal, while smoke related DNA adducts in the heart were minimally inhibited by high-dose oltipraz. In a repeat experiment that employed a 3-fold lower dose of cigarette smoke, oltipraz (500 p.p.m.) was found to inhibit the formation of DNA adducts in rat lungs and trachea by 80 and 65%, respectively. These data clearly demonstrate a high efficacy of oltipraz in inhibiting the formation of cigarette smoke-induced DNA adducts in the target tissues. PMID- 9744552 TI - An evaluation of the cytochrome P450 induction potential of pantoprazole in primary human hepatocytes. AB - Primary human hepatocytes contain a full complement of human drug-metabolizing enzymes and therefore represent a relevant experimental system for the evaluation of pharmacokinetic drug-drug interaction potential in human. In this study, the cytochrome P450 (CYP) induction potential of pantoprazole (PAN) was evaluated and compared to two other proton pump inhibitors (PPIs), omeprazole (OM) and lansoprazole (LAN). Primary human hepatocytes from three donors were studied. The hepatocytes were cultured for 3 days, followed by treatment for 3 days with the PPIs at 2, 5 and 10 microM. Two other known CYP inducers, 3-methylcholanthrene at 1 microM and rifampin at 50 microM, were also evaluated. Induction potentials of these chemicals for CYP1A and CYP3A were evaluated by isozyme activity and isozyme content. 7-Ethoxyresorufin-O-deethylase and testosterone 6beta hydroxylase activities were used as endpoints for CYP1A and CYP3A, respectively. Isozyme protein contents of CYP1A and CYP3A were evaluated via Western blotting. The results showed that for CYP1A induction, the rank ordering in induction potential was consistently OM > LAN > PAN. CYP3A induction by the PPI's were observed in two of the three hepatocyte cultures, with no apparent differences in induction potency for the three compounds. Our results on CYP1A induction suggest that PAN has a lower drug-drug interaction potential than OM and LAN. PMID- 9744553 TI - Determination of folate transport pathways in cultured rat proximal tubule cells. AB - Deficiency of the vitamin folic acid has recently been linked with increased incidence of neural tube defects and of cardiovascular disease, through elevated plasma homocysteine levels. The kidney has an important role in conserving folate to counteract development of deficiency. Urinary folate excretion is regulated by the degree of reabsorption of folate by the proximal tubule cell. To evaluate an in vitro model for studies of the regulation of urinary folate excretion, the present studies examined the transport of 5-methyltetrahydrofolate (5-CH3 H4PteGlu), the primary form of folate in the glomerular filtrate, by normal rat proximal tubule (RPT) cells in confluent monolayer cultures. Specific binding of 5-CH3-H4PteGlu to the apical membrane was saturable (K(D) = 27 nM), but intracellular transport was not saturated up to 100 nM concentrations. 5-CH3 H4PteGlu transport was decreased 50% by concentrations of folic acid that completely blocked 5-CH3-H4PteGlu binding by the apical folate receptor. Probenecid (10 mM), an anion exchange (reduced folate carrier) inhibitor, reduced 5CH3-H4PteGlu transport by 50% without significantly affecting binding. Aspirin (3 mM) did not alter 5-CH3-H4PteGlu transport, but significantly enhanced the inhibition due to probenecid. Similarly, indomethacin (5 microM) potentiated the inhibition of 5-CH3-H4PteGlu transport by probenecid. These data suggest that RPT cells take up 5-CH3-H4PteGlu by both the folate receptor and the reduced folate carrier, implying a role for both pathways in regulating urinary folate excretion. PMID- 9744554 TI - Ethanol-induced effects on expression level, activity, and distribution of protein kinase C isoforms in rat liver Golgi apparatus. AB - Acute ethanol administration induces significant modifications both in secretive and formative membranes of rat liver Golgi apparatus. The decrease in glycolipoprotein secretion and their retention into the hepatocyte contribute to the pathogenesis of alcohol-induced fatty liver. Molecular and cellular mechanisms behind the ethanol-induced injury of the liver secretory pathway are not yet completely defined. In this study on intact livers from ethanol-treated rats, the involvement of the Golgi compartment in the impairment of hepatic glycolipoprotein secretion has been correlated with changes in the expression level, subcellular distribution and enzymatic activity of protein kinase C (PKC) isoforms. Acute ethanol exposure determined a translocation of classic PKCs and delta isoform from the cytosol to cis and trans Golgi membranes, the site of glycolipoprotein retention in the hepatic cell. A marked stimulation of cytosolic epsilon PKC activity was observed throughout the period of treatment. The presence of activated PKC isozymes at the Golgi compartment of alcohol-treated rat livers may play a role in hepatic secretion and protein accumulation. Direct and indirect effects of ethanol consumption on PKC isozymes and Golgi function are discussed. PMID- 9744555 TI - Antioxidant activity of gallates: an electrochemical study in aqueous media. AB - The electron-donating ability of gallates, which are food and pharmaceutical antioxidants, is quantitatively assessed on the basis of their electrochemical characteristics. Gallic acid and the propyl, i-propyl, butyl, i-butyl, pentyl and i-pentyl gallate derivatives were electrochemically oxidized on the glassy carbon electrode by using differential pulse voltammetry, cyclic voltammetry and hydrodynamic voltammetry on the rotating disk electrode. All the compounds under study were easily oxidized in acidic and neutral solutions. Electrochemical oxidation occurs via two electron-transfer steps; however good resolution for the second wave was obtained only by using hydrodynamic conditions. The oxidation process results to be irreversible, diffusion controlled and pH-dependent. The introduction of the alkyl groups seems to affect the intensities of the semiquinone gallate radicals as can be ascribed from the observed differences in i(II)d/i(I)d ratio obtained from hydrodynamic voltammetric experiments for the different derivatives. We have found that the intensity of the gallate radicals follows the sequence GA > or = i-PG > PG > i-BG > BG > i-PeG > PeG. From the pH dependence of the peak current it is possible to affirm that pH 2 is the better condition for the oxidative activity showing that the antioxidant behaviour of these compounds are important in the stomach acid. PMID- 9744556 TI - Kinetic analysis in living cells of the inhibition of the P-glycoprotein-mediated efflux of anthracyclines by vinca alkaloids. AB - Cells that overexpress the mdr 1 gene have decreased steady-state accumulation and increased efflux of many anticancer drugs including anthracyclines and vinca alkaloids. The mechanism(s) of P-glycoprotein-mediated efflux of drugs is (are) still poorly understood. In an attempt to identify mechanism(s) by which multidrug resistance can be circumvented, the cellular accumulation has been examined of pirarubicin, doxorubicin and idarubicin alone and in conjunction with four vinca alkaloid derivatives--vinblastine, navelbine, vindesine and vincristine. The present study was performed using a spectrofluorometric method with which it is possible to follow continuously the uptake and release of fluorescent molecules by living cells, as the incubation of the cells with the drug proceeds. Erythroleukemia K562 cell lines were used. It has been shown that the P-glycoprotein-mediated efflux of these three anthracyclines can be inhibited by vinca alkaloids derivatives. At pH 7.2, 50% of the P-glycoprotein-mediated efflux of daunorubicin and idarubicin was inhibited by about 40 +/- 10 microM vinblastine and that of pirarubicin by 10 +/- 2 microM vinblastine. The vinblastine concentration required to inhibit 50% of the active efflux of these anthracyclines did not depend on the anthracycline concentrations used, indicating that the inhibition was non competitive. The ability of navelbine, vincristine and vindesine to inhibit the active efflux of pirarubicin was also checked; 15 +/- 3 microM navelbine are required to inhibit 50% of the active efflux but at concentrations lower than 100 microM, neither vincristine nor vindesine were able to inhibit this efflux, indicating that the vinca alkaloids compounds which are the most efficient are the most lipophilic. For the four vinca alkaloids, the concentration required to inhibit 50% of the efflux was lower as the pH was higher. A detailed kinetics analysis of the P-glycoprotein mediated efflux of pirarubicin in the presence of vinblastine indicates a non competitive inhibition with K(I) = 12 +/- 2 microM. PMID- 9744558 TI - Effects of beta-ionone on the expression of cytochrome P450s and NADPH-cytochrome P450 reductase in Sprague Dawley rats. AB - We have recently reported that beta-ionone induces cytochrome P450 (P450) 2B1 in rats. Effects of beta-ionone on the expression of other P450 isozymes and NADPH P450 reductase were further investigated in Sprague Dawley rats. Administration of beta-ionone subcutaneously 72 and 48 h before sacrificing the animals not only significantly induced the liver microsomal activities of P450-associated enzymes and NADPH-P450 reductase, but also clearly increased in the level of P450 1A1/2, P450 2C, and NADPH-P450 reductase proteins. The induction of P450 1A1/2 and 2C by beta-ionone was much greater in male than in female as measured by western immunoblotting. Reverse transcriptase-polymerase chain reactions showed that, in addition to P450 2B1 and 2B2 mRNAs, P450 1A2, 2C6 and NADPH-P450 reductase mRNAs were increased when beta-ionone was administered. Our previous and present results indicated that beta-ionone may induce several P450s and NADPH-P450 reductase by the accumulation of their corresponding mRNAs. PMID- 9744557 TI - Metabolism and activation of cyclopenta polycyclic aromatic hydrocarbons in isolated human lymphocytes, HL-60 cells and exposed rats. AB - The metabolism of radiolabelled benz(j)aceanthrylene (B(j)A) was studied in suspensions of isolated human peripheral mononuclear blood cells (lymphocytes), using high performance liquid chromatography (HPLC). The only known metabolite found after 24 h exposure to 30 microg/ml (120 microM) B(j)A, was B(j)A-1,2 dihydrodiol, representing approximately 35% of the total metabolites formed. B(j)A, benz(l)aceanthrylene (B(l)A) and benzo(a)pyrene (B(a)P) all caused DNA adducts in human lymphocytes, as well as in the human promyelocytic cell line HL 60 cells, as measured by the 32P-postlabelling technique (30 microg/ml, 24 h). The total DNA adduct levels in human lymphocytes exposed to B(j)A, B(l)A or B(a)P were 0.13 +/- 0.03, 1.10 +/- 0.62 and 0.37 +/- 0.10 fmol/microg DNA, respectively, and similar levels were obtained in HL-60 cells (0.18 +/- 0.14, 0.97 +/- 0.35 and 0.29 +/- 0.17 fmol/microg DNA, respectively). For each compound, the human lymphocytes and HL-60 cells in addition showed similar DNA adduct patterns. Cells exposed to B(j)A revealed only one DNA adduct, which, judged by its TLC properties, resulted from B(j)A-1,2-epoxide. As measured by the alkaline filter elution technique, only low levels of single strand DNA breaks (SSB) were observed in both human lymphocytes and HL-60 cells after exposure to B(j)A, B(l)A or B(a)P (24 h, 30 microg/ml). By adding cytosine-1-beta-D arabinofuranoside (Are C) and hydroxyurea (HU) 1 h prior to analysis to prevent strand break rejoining, some increase in SSB (2-3 times) was observed in the lymphocytes. Co-incubation of human lymphocytes with liver microsomes from PCB treated rats for 1 h and exposure to B(j)A or B(l)A, increased the DNA adduct levels in the lymphocytes to 12.3 and 37.8 fmol/microg DNA, respectively. A large increase in SSB was also observed, whereas no such increase was observed after co incubation with human liver microsomes. In vivo exposure of rats to 30 mg/kg B(j)A (i.p.) for 24 h revealed one major DNA adduct in lymphocytes and lung tissue (only one of three rats showed an adduct in liver tissue), apparently resulting from B(j)A-1,2-epoxide. The total DNA adduct level in the lymphocytes was 0.09 fmol/microg DNA, and in lung tissue between 0.10 and 0.62 fmol/microg DNA. Overall, the present data suggests that oxidation at the cyclopenta-ring is an important activation pathway for B(j)A in rats as well as in humans. PMID- 9744559 TI - Inhibition of linoleic acid hydroperoxide-induced toxicity in cultured human umbilical vein endothelial cells by catechins. AB - The protective effect of catechins, major components of green tea, was studied in cultured human umbilical vein endothelial cells exposed to toxicity induced by linoleic acid hydroperoxide (LOOH). In the case where cells were incubated in medium containing both LOOH and catechins, (+)-catechin (C) was effective in suppressing of LOOH-induced cytotoxicity, but (-)-epicatechin (EC), (-) epigallocatechin (EGC), (-)-epicatechin gallate (ECG), and (-)-epigallocatechin gallate (EGCG) had no effect. EGCG monoglucoside (EGCG-G1) and EGCG diglucoside (EGCG-G2), apophilic derivatives of EGCG, show a protective effect on LOOH induced cytotoxicity when present at the time of treatment with LOOH. On the other hand, when cells were incubated with catechins for 24 h before treatment with LOOH there was no protection against the oxidative damage by LOOH. Furthermore, the interaction between catechins and alpha-tocopherol was examined under these culture conditions. C showed a synergistic effect with alpha tocopherol in protecting against LOOH-induced damage. These results suggest that catechins interact with LOOH present in the medium or near the surface of membranes, but not with LOOH incorporated into cellular membranes and that catechins are able to interact with alpha-tocopherol to provide synergistic protection against the cytotoxicity of LOOH. PMID- 9744560 TI - Inhibition of eukaryote protein kinases and of a cyclic nucleotide-binding phosphatase by prenylated xanthones. AB - A series of prenylated xanthones are variously potent inhibitors of the catalytic subunit (cAK) of rat liver cyclic AMP-dependent protein kinase (PKA), rat brain Ca2+ and phospholipid-dependent protein kinase C (PKC), chicken gizzard myosin light chain kinase (MLCK), wheat embryo Ca2+-dependent protein kinase (CDPK) and potato tuber cyclic nucleotide-binding phosphatase (Pase). The prenylated xanthones examined are mostly derivatives of alpha-mangostin in which the 3 hydroxyl and 6-hydroxyl are variously substituted with groups R or R', respectively, or derivatives of 3-isomangostin (mangostanol) in which the 9 hydroxyl is substituted with groups R' or the prenyl side chain is modified. The most potent inhibitors of cAK have non-protonatable and relatively small R' and R groups. Conversely, the most potent inhibitors of PKC and MLCK have bulkier and basic R' groups. Some prenylated xanthones are also potent inhibitors of CDPK. PKC and cAK are competitively inhibited by particular prenylated xanthones whereas the compounds that are the most potent inhibitors of MLCK and CDPK are non-competitive inhibitors. Prenylated xanthones having relatively small and non protonatable R' and R groups inhibit a high-affinity cyclic nucleotide binding Pase in a non-competitive fashion. PMID- 9744561 TI - Signaling from G-protein-coupled receptors to mitogen-activated protein (MAP) kinase cascades. AB - Heterotrimeric GTP-binding protein (G-protein)-coupled receptors are able to induce a variety of responses including cell proliferation, differentiation, and activation of several intracellular kinase cascades. Prominent among these kinases are the activation of mitogen-activated protein (MAP) kinase, including the extracellular signal-regulated kinases (ERKs), ERK1 and ERK2 (p44mapk and p42mapk, respectively); stress-activated protein kinases (SAPKs/JNKs); and p38 kinase. These receptors signal through G-proteins. Recent data have shown that the activation of mitogen-activated protein/ERK kinase induced by G-protein coupled receptors is mediated by both Galpha and Gbetagamma subunits involving a common signaling pathway with receptor-tyrosine-kinases. Gbetagamma-mediated mitogen-activated protein kinase activation is mediated by activation of phosphoinositide 3-kinase, followed by a tyrosine phosphorylation event, and proceeds in a sequence of events that involve functional association among the adaptor proteins Shc, Grb2, and Sos. SAPKs/JNKs and p38 are able to be activated by Gbetagamma proteins in a pathway involving Rho family proteins including RhoA, Rac1, and Cdc42. PMID- 9744562 TI - Oxidized low-density lipoprotein, a two-faced Janus in coronary artery disease? AB - The word antioxidant has become a household term, and every day we are bombarded with claims of antioxidant protection against a host of diseases. Atherosclerosis, cancer, gastric ulcers, memory loss, rheumatoid arthritis, endometriosis, pregnancy complications, hypertension, stroke, and a host of other diseases have been suggested to be induced by oxidative stress, and antioxidants have been suggested to be beneficial in the prevention and treatment of these disorders. While some of these may be exuberant claims, atherosclerosis is one disease in which the oxidation hypothesis has taken firm roots. The oxidation of low-density lipoprotein (LDL) has been suggested to be a key step in the initiation of the early atherosclerotic lesion. A number of proatherogenic effects have been described for both the protein and lipid components of oxidized low-density lipoprotein. In this commentary, a brief description of the involvement of oxidation and the potential for antioxidant treatment for cardiovascular disease will be provided. However, there are innumerable questions plaguing the hypothesis; this commentary, therefore, will also serve as a devil's advocate and propose that some form of oxidation might actually be beneficial. PMID- 9744563 TI - Kappa-opioid modulation of human microglial cell superoxide anion generation. AB - Opioids have been postulated to play an immunomodulatory role in the CNS. Recently, we found that priming microglia with interferon (IFN)-gamma or tumor necrosis factor (TNF)-alpha resulted in an enhanced production of superoxide anion, a reactive oxygen intermediate that may be pathogenic during brain inflammation. In the present study, we investigated the effects of trans-3,4 dichloro-N-methyl-N[2-(1-pyrolidinyl)cyclohexyl]benze neaceamide methanesulfonate (U50,488), a selective kappa-opioid ligand, on microglial cell superoxide production when cells were primed with cytokines or stimulated with phorbol myristate acetate. While treatment of microglial cells with U50,488 had little effect on nonstimulated or stimulated superoxide production, this opioid inhibited (by >70%) the priming effects of cytokines. Maximal inhibition of microglial cell superoxide generation by U50,488 was observed at 10 nM for the priming effect of interferon-gamma and at 1 microM for tumor necrosis factor alpha. Pretreatment of microglial cell cultures for 30 min with an equal concentration of the selective kappa-opioid receptor antagonist nor binaltorphimine (nor-BNI) completely blocked the inhibitory effect of U50,488. The results of this study suggest that kappa-opioids may have therapeutic potential in inflammatory diseases of the CNS involving reactive oxygen intermediates produced by activated microglia. PMID- 9744564 TI - Delta-opioid suppression of human immunodeficiency virus-1 expression in T cells (Jurkat). AB - Delta-opioid receptor (DOR) transcripts and binding sites are expressed by lymphocytes and lymphoid cell lines from several species. Direct modulation of lymphocyte function through DORs affects T cell proliferation, interleukin-2 production, chemotaxis, and intracellular signaling. Moreover, in human DOR transfected T cells (DOR-Ju.1), delta-opioids have been shown previously to mobilize intracellular calcium rapidly, to inhibit forskolin-stimulated cyclic AMP production, and to activate the mitogen-activated protein kinases ERKs 1 and 2. These observations led us to consider whether delta agonists modify T cell functions, thus affecting the expression of human immunodeficiency virus-1 (HIV 1) by CD4+ T cells. To test this hypothesis, DOR-Ju.1 cells, derived from Jurkat cells stably transfected with a cDNA encoding the neuronal DOR, were stimulated with deltorphin or benzamide, 4-[[2,5-dimethyl-4-(2-propenyl)-1-piperazinyl](3 methoxyphenyl)methyl]N- ,[2S[(S*),2alpha,5beta]]-(9Cl) (SNC-80) prior to the addition of HIV-1. Both deltorphin and SNC-80 concentration-dependently inhibited the production of p24 antigen, an index of HIV-1 expression. Inhibition was maximal with 10(-13)-10(-9) M SNC-80 (>60% reduction) or 10(-15)-10(-11) M deltorphin (>50% reduction). At higher concentrations, less inhibition of p24 antigen production was found. Naltrindole (NTI, 10(-11) M), a selective DOR antagonist, abolished the inhibitory effects of 10(-9) M SNC-80, whereas 10(-13) M NTI partially reversed the effect of SNC-80. Thus, activation of DORs expressed by CD4+ T cells significantly (P < 0.05) reduced the expression of HIV-1 by these cells. These findings suggest that opioid immunomodulation directed at host T cells may be adjunctive to standard antiviral approaches to HIV-1 infection. PMID- 9744565 TI - Inhibition of acetylcholinesterase and butyrylcholinesterase by chlorpyrifos oxon. AB - Phosphorothionate insecticides such as parathion (O,O-diethyl O-p-nitrophenyl phosphorothioate) and chlorpyrifos (CPS; O,O-diethyl O-3,5,6-trichloro-2-pyridyl phosphorothioate; Dursban) are metabolically converted by oxidative desulfuration into paraoxon and chlorpyrifos-oxon (CPO). The insecticidal action of chlorpyrifos stems from inhibition of acetylcholinesterase (AChE) by CPO, resulting in severe cholinergic toxicity. Sensory peripheral neuropathy was observed in people exposed environmentally to chlorpyrifos sprayed in confined areas. We have examined the kinetics of inhibition of AChE and butyrylcholinesterase (BChE) by paraoxon and CPO. The bimolecular rate constants (ki) for inhibition by paraoxon of recombinant human (rH) AChE, recombinant mouse (rM) AChE, and fetal bovine serum (FBS) AChE were 7.0, 4.0, and 3.2 x 10(5) M(-1) min(-1). The ki values for the inhibition by CPO of rH AChE, fetal bovine serum AChE, human RBC AChE, Torpedo AChE, and recombinant mouse (rM) AChE were 9.3, 2.2, 3.8, 8.0, and 5.1 x 10(6) M(-1) min(-1), respectively. Inhibition of human serum BChE, rH BChE, and rM BChE by CPO yielded ki values of 1.65, 1.67, and 0.78 x 10(9) M(-1) min(-1), respectively. The ki values obtained for BChE from various species were 160- to 750-fold larger than those of AChE from parallel sources. Inhibition of the single-site mutant A328Y of rH BChE by CPO displayed a 21-fold lower rate than that of wild-type rH BChE (ki, 7.9 x 10(7) vs 1.67 x 10(9) M(-1) min(-1)). The double mutant of acyl pocket residues of rH AChE, F295L/F297V, was inhibited by CPO with a 150-fold larger ki than wild type (1.5 x 10(9) vs 1.0 x 10(7) M(-1) min(-1)). The increased rate obtained with the double mutant displaying characteristics of the BChE active center provides a rationale for higher efficacy of CPO scavenging by BChE, compared with AChE. PMID- 9744566 TI - Superoxide radical scavenging by phenolic bronchodilators under aprotic and aqueous conditions. AB - Asthmatic airway disease is accompanied by the appearance of inflammatory cells which produce reactive oxygen species (ROS). Therefore, the radical scavenging properties of the bronchodilators reproterol, fenoterol, salbutamol and terbutaline toward superoxide anion radicals and hydroperoxyl radicals were investigated in a model system by electron paramagnetic resonance spectroscopy (EPR) and photometric approaches. The substances under study showed activity in superoxide radical scavenging under aprotic and protic conditions as well. The efficiency of the reaction decreased in the order: fenoterol > salbutamol > reproterol > terbutaline > oxyfedrine when DMSO was used as an aprotic solvent. In an aqueous system, the rate constants decreased in the order: fenoterol > reproterol > salbutamol. It is suggested that the antioxidant effect of these beta2-agonists is an additional advantage in treatment of asthmatic lung disease, reducing the negative consequences of airway inflammation. PMID- 9744567 TI - The activation of gold complexes by cyanide produced by polymorphonuclear leukocytes. III. The formation of aurocyanide by myeloperoxidase. AB - There is considerable evidence that the anti-rheumatic gold complexes are activated by their conversion to aurocyanide. In order to understand the mechanism of production of aurocyanide, we investigated the involvement of myeloperoxidase in the reaction. This haem enzyme of neutrophils and monocytes uses hydrogen peroxide to oxidise chloride and thiocyanate to hypochlorous acid and hypothiocyanite, respectively. When aurothiomalate (10 microM) was incubated with thiocyanate (200 microM), hydrogen peroxide (100 microM) and myeloperoxidase (20 nM), it was transformed to a product that was spectrally identical to authentic aurocyanide. Aurothiomalate was quantitatively converted to aurocyanide in about 10 min at pH 6.0 and in 40 min at pH 7.4. Aurocyanide formation occurred after myeloperoxidase had used all the hydrogen peroxide available to produce hypothiocyanite. Thus, the cyanide must have formed from the slow decomposition of hypothiocyanite. The rate of aurocyanide production was increased in the presence of 100 mM chloride, which indicates that hypochlorous acid accelerates the formation of cyanide. Hypochlorous acid (100 to 400 microM) reacted non enzymatically with thiocyanate (200 microM) and aurothiomalate (10 microM) to produce aurocyanide. Thus, aurocyanide is produced by two processes, involving both the formation of hypothiocyanite and hypochlorous acid. Aurocyanide is an effective inhibitor of the respiratory burst of neutrophils and monocytes and the proliferation of lymphocytes. Therefore, aurothiomalate may attenuate inflammation by acting as a pro-drug which is reliant on neutrophils and monocytes to produce hypothiocyanite. When the hypothiocyanite decays to hydrogen cyanide, the pro-drug is converted to aurocyanide which then suppresses further oxidant production by these inflammatory cells. PMID- 9744568 TI - Role for the plasmodium falciparum digestive vacuole in chloroquine resistance. AB - We have developed a method for the isolation of pure and intact Plasmodium falciparum digestive vacuoles capable of ATP-dependent chloroquine (CQ) accumulation in vitro. The method is rapid and reliable, and it produces a high yield of vacuoles (20%). CQ accumulation in isolated vacuoles was found to be ATP , Mg2+-, and temperature-dependent. We then investigated the CQ-accumulating capabilities of vacuoles isolated from CQ-resistant (CQR) and CQ-sensitive (CQS) parasites. At external CQ concentrations of 100 and 250 nM, vacuoles isolated from two CQS strains (D10 and RSA3) (Vm: 380-424 fmol/10(6) vacuoles/hr) accumulated significantly more CQ (approximately 3 times) than those isolated from three (FAC8, RSA11, and RSA15) of the four CQ-resistant strains of P. falciparum tested (Vmax: 127-156 fmol/10(6) vacuoles/hr) (P < or = 0.05). We propose that the low level of CQ accumulation observed in vacuoles isolated from most of the CQ-resistant parasites tested contributes to the decreased CQ accumulation seen in these strains and, hence, to CQ resistance. Although it is often suggested that the digestive vacuole of the P. falciparum parasite is involved in the mechanism of CQ resistance, to our knowledge this is the first direct confirmation. PMID- 9744569 TI - Structure-activity relationships for triphenylethylene antiestrogens on hepatic phase-I and phase-II enzyme expression. AB - To better understand the mechanism(s) by which tamoxifen induces rat hepatic CYPIIB2 and suppresses GSTA1, structure-activity studies were performed. Compounds employed in these studies included: tamoxifen, fixed-ring tamoxifen, ethylated fixed-ring tamoxifen, pyrrolidino-tamoxifen, 4-iodotamoxifen, idoxifene, and toremifene. With respect to GSTA1 suppression, tamoxifen, fixed ring tamoxifen, 4-iodotamoxifen, idoxifene, and toremifene were all potent suppressors of GSTA1, while ethylated fixed-ring tamoxifen and pyrrolidino tamoxifen were completely without activity. The results suggest that the aminoethoxy side chain plays a crucial role in GSTA1 suppression, and that 4 iodination may potentiate this activity. With respect to induction of CYPIIB2, tamoxifen, fixed-ring tamoxifen, and ethylated fixed-ring tamoxifen were inducers of this enzyme, while toremifene and 4-iodotamoxifen were inactive, suggesting that the aminoethoxy side chain is not a structural determinant of CYPIIB2 induction. Because ethylated fixed-ring tamoxifen, toremifene, and 4 iodotamoxifen had differential activities in the two assays, we conclude that CYPIIB2 induction and GSTA1 suppression by triphenylethylenes are the result of two separate and distinct mechanistic pathways. Structure-activity relationships for GSTA1 suppression and CYPIIB2 induction were compared with previously published relationships for triphenylethylene: 1) estrogen receptor relative binding affinity; 2) calmodulin antagonism; 3) antiuterotrophic activity; and 4) antagonism of MCF-7 cell growth. No clear correlation was observed between the effects on CYPIIB2 and these other four activities, suggesting no relationship between the mechanisms responsible for these effects. Similarly, no precise correlation was observed between GSTA1 suppression and these other activities, although rough similarities were observed for relative binding affinity and antiuterotrophic activity. This suggests that the mechanisms responsible for CYPIIB2 induction and GSTA1 suppression are not related to the mechanisms of action for these other documented activities, and may represent different mechanistic pathways. PMID- 9744570 TI - Unique inhibitory action of the synthetic compound 2-[N-(2-aminoethyl)-N-(5 isoquinolinesulfonyl)] amino-N-(4-chlorocinnamyl)-N-methylbenzylamine (CKA-1306) against calcium/calmodulin-dependent protein kinase I. AB - A newly synthesized compound, 2-[N-(2-aminoethyl)-N-(5 isoquinolinesulfonyl)]amino-N-(4-chlorocinnamyl )-N-methylbenzylamine (CKA-1306), was found to inhibit cyclic AMP-dependent protein kinase (PKA) and Ca2+/calmodulin-dependent protein kinase I (CaMK I) with IC50 values of 1.6+/ 0.14 and 2.5+/-0.16 microM, respectively. In contrast, the established PKA inhibitors H-8 and H-89 inhibited CaMK I with relatively high IC50 values of >100 and 24.4+/-3.2 microM, respectively. An additional inhibitor, KN-62, against Ca2+/calmodulin-dependent protein kinase II (CaMK II) did not inhibit either PKA or CaMK I at the concentrations tested. In our library of many isoquinolinesulfonamide derivatives, only CKA-1306 inhibited CaMK I to a satisfactory degree, suggesting a unique mode of action. Indeed, the inhibition of CaMK I by CKA-1306 was competitive in every respect to Mg2+/ATP, peptide substrate (syntide-2), and Ca2+/calmodulin. This phenomenon may be understood from the context of the recently determined structure of the enzyme in its autoinhibited state. Such kinetic analysis was also extended to cases using a phosphorylated and activated enzyme at Thr177 or a constitutively active, COOH terminal truncated mutant at Gln293. CKA-1306 still competed with Mg2+/ATP for the two enzymes, but it no longer achieved any competitive advantage over syntide 2. These results may reflect some differences in the active conformation of CaMK I. However, the compound should be constant in its recognition of an Mg2+/ATP binding site of the enzyme. Though CKA-1306 is not specific to CaMK I, the compound will be useful in studying the enzyme further under limited conditions. PMID- 9744571 TI - Oxidative stress and 1-methyl-2-nitroimidazole cytotoxicity. AB - The 2-nitroimidazoles have been used clinically to radiosensitize resistant hypoxic cells, but a dose-limiting peripheral neuropathy has restricted their therapeutic effectiveness. A model compound, 1-methyl-2-nitroimidazole (INO2), was used to investigate the possible role of oxidative stress in this normal tissue toxicity. Chinese hamster ovary (CHO) cells were 10-15 times more resistant to 20 mM INO2 under aerobic than hypoxic conditions. In comparison, a pair of transformed rat embryo fibroblasts (ER17-1wtp53 and ER12L5mtpP53), differing in their p53 genotype, were approximately 3- to 4-fold more sensitive than Chinese hamster ovary cells to INO2 under aerobic conditions, but had the same sensitivity as Chinese hamster ovary cells under hypoxic conditions. These results are consistent with an earlier hypothesis that the mechanism of aerobic toxicity is different from that of hypoxic toxicity (nitroreduction) and show that neither toxicity is dependent on cellular p53 status. There was an increase in the production of reactive oxygen intermediates and a decrease in the antioxidant glutathione following aerobic exposure to INO2, which correlated with cell survival in all three cell lines. No evidence of reductive adducts of the 2 nitroimidazole 2-(2-nitro-1H-imidazol-1-yl)-N-(2,2,3,3,3-pentafluoropropyl)acetam ide (EF5) was found by immunofluorescent techniques in aerobic cells. Differing activities of the antioxidant enzymes superoxide dismutase and catalase could be correlated with INO2 aerobic cytotoxicity. DNA strand breaks, as measured by the comet assay, paralleled the appearance of INO2 aerobic cytotoxicity in all three cell lines. Taken together, these results strongly support the conclusion that the aerobic toxicity of IN02 is due to active oxygen species created by futile redox cycling of the parent compound. PMID- 9744572 TI - Tumor cell resistance to topoisomerase II poisons: role for intracellular free calcium in the sensitization by inhibitors or calcium-calmodulin-dependent enzymes. AB - Tumor cell resistance to inhibitors of topoisomerase II (topo II) is associated frequently with the overexpression of P-glycoprotein (PGP), and strategies to overcome resistance are focused on restoring defects in drug accumulation. Inhibitors of calcium-calmodulin-dependent enzymes sensitize resistant tumor cells to the topo II poison etoposide (VP-16) by enhancing DNA damage and an apoptotic response. In the present study, we have investigated the consequences of buffering intracellular calcium with 1,2-bis(o-aminophenoxy)ethane-N,N,N'N' tetraacetic acid tetra(acetoxy-methyl) ester (BAPTA-AM) on the sensitizing effects of the calmodulin-dependent protein kinase II inhibitor 1-[N,O-bis(1,5 isoquinolinesulfonyl)-N-methyl-L-tyrosyl]-4-piperazine (KN-62) in etoposide resistant human leukemia HL-60 (HL-60/ADR0.05) cells. In cells pretreated with 20 microM BAPTA-AM for 2 hr, extracellular ATP failed to trigger intracellular calcium transients, and no effects on the accumulation of VP-16 were apparent. Also, the effect of KN-62 in significantly (P=0.002 to 0.042) enhancing the accumulation of VP-16 in HL-60/ADR0.05 cells was unaffected due to pretreatment with BAPTA-AM. In contrast, pretreatment with BAPTA-AM reduced the DNA damage induced by VP-16, and significantly (P=0.038) reversed the enhancement by KN-62 of VP-16-stabilized topo II-mediated DNA cleavable complex formation. The pretreatment of HL-60/ADR0.05 cells with BAPTA-AM was also associated with the hypophosphorylation of topo IIalpha. Consistent with the ability of BAPTA-AM to circumvent the potentiation by KN-62 of VP-16-induced DNA damage, survival of cells treated with 40 microM VP-16 in the absence of KN-62 and 10 microM VP-16 in the presence of KN-62 was significantly (P=0.026 to 0.031) higher due to BAPTA-AM pretreatment. Results demonstrate that intracellular calcium transients could play a key role in the sensitization of etoposide-resistant tumor cells by inhibitors of calcium-calmodulin-dependent enzymes. PMID- 9744573 TI - Cytotoxic and DNA-damaging properties of N-[2-(dimethylamino)ethyl]acridine-4 carboxamide (DACA) and its analogues. AB - An antitumor drug N-[2-(dimethylamino)ethyl]acridine-4-carboxamide (DACA) and its three close structural analogs N-[2-(hydroxyethylamino)ethyl]acridine-4 carboxamide (DACAH), N-[2-(dimethylamino)ethyl]-9-aminoacridine-4-carboxamide (amino-DACA), and N-[2-(hydroxyethylamino)ethyl]-9-aminoacridine-4-carboxamide (amino-DACAH) were studied for their ability to inhibit RNA synthesis in vitro and to form topoisomerase II-mediated DNA lesions in relation to cell-killing activity. All tested compounds induced chromatin lesions characteristic of topoisomerase II-blocking drugs (DNA breaks and DNA-protein cross-links) in treated cells, but were much less active than reference antileukemic acridine m AMSA (4'-(9-acridinylamino)-methanesulfon-m-anisidide). The ability to form these lesions was dependent on the structure of the 4-carboxamide side-chain, which seems to be an important factor affecting the drug transport rate through cell membrane. A 4-carboxamide chain with an N-2-(dimethylamino)ethyl moiety resulted in more efficient transport through cell membranes, higher cytotoxicity, and DNA damaging activity. The mode of action of acridine-4-carboxamides was further elucidated by their incubation with cells in the presence of antitopoisomerase II agents of a known mechanism of inhibition. These were: bisdioxopiperazine (ICRF 187), a catalytic inhibitor of topoisomerase II, and etoposide (VP-16), an inducer of a cleavable complex of the enzyme with DNA. The cytotoxicity of DACA and its analogs was not antagonized by preincubating cells with ICRF-187. All tested acridines protected cells against DNA breakage induced by VP-16, but the extent of protection varied significantly. Amino-DACA, which easily penetrates cell membrane, fully inhibited DNA break formation, whereas other analogs exhibited a low degree of protection when used at high concentration. Our results suggest that the acridine-4-carboxamides discussed here are poor topoisomerase II poisons and that this enzyme is not their main target. PMID- 9744574 TI - Oxidative DNA damage and apoptosis induced by metabolites of butylated hydroxytoluene. AB - DNA damage by metabolites of a food additive, butylated hydroxytoluene (BHT), was investigated as a potential mechanism of carcinogenicity. The mechanism of DNA damage by 2,6-di-tert-butyl-p-benzoquinone (BHT-quinone), 2,6-di-tert-butyl-4 hydroperoxyl-4-methyl-2,5-cyclohexadienone (BHT-OOH), and 3,5-di-tert-butyl-4 hydroxybenzaldehyde (BHT-CHO) in the presence of metal ions was investigated by using 32P-labeled DNA fragments obtained from the c-Ha-ras-1 proto-oncogene and the p53 tumor suppressor gene. BHT-OOH caused DNA damage in the presence of Cu(II), whereas BHT-quinone and BHT-CHO did not. However, BHT-quinone did induce DNA damage in the presence of NADH and Cu(II). Bathocuproine inhibited Cu(II) mediated DNA damage, indicating the participation of Cu(I) in the process. Catalase also inhibited DNA damage induced by BHT-quinone, but not that induced by BHT-OOH. The DNA cleavage pattern observed with BHT-quinone plus NADH was different from that seen with BHT-OOH. With BHT-quinone plus NADH, piperidine labile sites could be generated at nucleotides other than adenine residue. BHT OOH caused cleavage specifically at guanine residues. Pulsed field gel electrophoresis showed that BHT-OOH and BHT-quinone induced DNA strand breaks in cultured cells, whereas BHT-CHO did not. Both BHT-quinone and BHT-OOH induced internucleosomal DNA fragmentation, which is the characteristic of apoptosis. Furthermore, flow cytometry analysis revealed an increase of peroxides in cultured cells treated with BHT-OOH or BHT-quinone. These results suggest that BHT-OOH participates in oxidative DNA damage directly, whereas BHT-quinone causes DNA damage through H2O2 generation, which leads to internucleosomal DNA fragmentation. PMID- 9744575 TI - Expression of multiple forms of cytochrome P450 and associated mono-oxygenase activities in rat brain regions. AB - Cytochrome P450 (P450) content and P450-mediated mono-oxygenase activities were measured in microsomes prepared from various regions of rat brain. The regional P450 content in brain varied between 0.1 and 0.15 nmol/mg of protein, with the brainstem and cerebellum showing the highest levels. NADPH cytochrome c reductase activity was highest in the cortex followed by cerebellum and brainstem as compared with the whole brain. Mono-oxygenase activities also varied among the various brain regions. Southern blot analysis of the cDNA synthesized from the poly(A)RNA isolated from rat brain regions and hybridized with cDNA to rat liver P4502B or P4502E1 revealed the presence of a transcript in untreated rat brain that had a molecular mass similar to that of the corresponding transcript from rat liver. Immunoblot analyses using antisera to purified rat liver P4502E1, P450(2B1/2B2), and a phenobarbital-inducible form of rat brain P450 revealed the presence of corresponding immunoreactive protein bands in all the brain regions examined. The present study demonstrated the diversity in the distribution of P450 and associated mono-oxygenase activities in brain and thus may reflect the differential capability of various regions of the brain to detoxify or bioactivate diverse xenobiotics. PMID- 9744576 TI - Development of a comprehensive panel of antibodies against the major xenobiotic metabolising forms of cytochrome P450 in humans. AB - Mono-specific antibodies against the human cytochrome P450 (P450) enzymes CYP1A1, CYP1A2, CYP1B1, CYP2A6, CYP2B6, CYP2D6, CYP2E1, CYP3A4, CYP3A5 and CYP4A11 and an antibody that binds to CYP2C8, CYP2C9 and CYP2C19 have been produced by immunising rabbits with synthetic peptides representing small regions of each of these P450 enzymes. The specificity of the antibodies was confirmed by immunoblotting using recombinant P450 enzymes and samples of human hepatic microsomal fraction. Each of the antibodies bound only to their respective target P450 enzyme(s). The relative intensity of immunoreactive bands was compared with a variety of P450 activities and correlations were found between CYP1A2 and phenacetin O-deethylase activity, CYP2A6 and coumarin 7-hydroxylase activity, CYP2C9 and tolbutamide 4-hydroxylase activity, CYP2C19 and S-mephenytoin 4 hydroxylase activity, CYP2D6 and debrisoquine 4-hydroxylase activity, CYP2E1 and chlorzoxazone 6-hydroxylase activity, CYP3A4 and midazolam 1'-hydroxylase activity, and CYP4A11 and lauric acid 12-hydroxylase activity. A proportion of the 30 liver samples examined lacked CYP2A6 (7%), CYP2C19 (10%) or CYP2D6 (13%), consistent with the polymorphic expression of these P450 enzymes in human liver. Although CYP3A5 was detected in most individuals (97%), expression was polymorphic with 20% containing substantially higher levels. CYP2B6 was expressed in 20% of the human liver samples, with one sample containing a particularly high level. No immunodetectable CYP1A1 or CYP1B1 was found, consistent with the low level of expression of these P450 enzymes in human liver. The results demonstrate the utility of the antipeptide approach for producing specific antibodies against human P450 enzymes, enabling a comprehensive panel of antibodies against human P450 enzymes to be produced. PMID- 9744577 TI - Decrease in thymidylate kinase activity in peripheral blood mononuclear cells from HIV-infected individuals. AB - Nucleosides and nucleoside analogs are anabolised to their triphosphates by intracellular kinases. The anti-HIV analogue zidovudine (AZT) is phosphorylated by cytosolic thymidine kinase 1 (TK1), thymidylate kinase (dTMPK), and nucleoside diphosphate kinase. It is known that dTMPK is one of the rate-limiting steps in the activation of zidovudine. The activities of TK1, dTMPK, and deoxycytidine kinase (dCK) were determined in extracts of in vitro activated peripheral blood mononuclear cells from HIV-infected patients and healthy noninfected individuals. dTMPK activity was 10-fold lower and TK1 activity was five-fold lower in extracts from infected as compared to uninfected persons. Deoxycytidine kinase activities in the extracts from both groups were very similar. Differences in in vitro activation, as determined by flow cytometry, of the peripheral lymphocytes were not responsible for the decreased TK1 and dTMPK activities. A reduced level of intracellular azido-dideoxythymidinetriphosphate in activated mononuclear cells from HIV-infected patients was also observed. The low levels of TK1 and dTMPK in lymphocytes from HIV-infected patients may be related to the anergy phenomenon observed as a result of HIV infection. This effect should also be considered in the development of new anti-HIV drugs. PMID- 9744578 TI - Kappa-opioid potentiation of tumor necrosis factor-alpha-induced anti-HIV-1 activity in acutely infected human brain cell cultures. AB - Opioids have been postulated to play an immunomodulatory role in the pathogenesis of HIV-1. Synthetic kappa-opioid receptor (KOR) ligands have been found to inhibit HIV-1 expression in acutely infected microglial cell cultures. We recently found that interleukin(IL)-1beta and tumor necrosis factor(TNF)-alpha have antiviral effects in acutely infected mixed glial/neuronal cell cultures. In the present study, we investigated whether selective KOR ligands would exert antiviral effects in acutely infected brain cell cultures. While the KOR ligand trans-3,4-dichloro-N-methyl-N[2-(1-pyrolidinyl)cyclohexyl]benze neaceamide methanesulfonate (U50,488) alone had little anti-HIV-1 activity, this opioid potentiated in a concentration-dependent manner the antiviral activity of TNF alpha, but not of IL-1beta. The potentiating effect of U50,488 was detected after a 6-hr pretreatment and peaked at 24 hr. The KOR antagonist nor-binaltorphimine completely blocked the potentiating effect of U50,488, suggesting the involvement of a KOR-mediated mechanism. Antibodies to TNF-alpha completely blocked the potentiating effect of U50,488, suggesting a critical role for TNF-alpha. Antibodies to IL-1beta blocked the potentiating effect of U50,488, suggesting that IL-1beta was released following U50,488 treatment, which might contribute to the potentiating effect of U50,488. These in vitro findings support the notion that synthetic kappa-opioids could be considered as potential adjunctive therapeutic agents in HIV-1-related brain disease. PMID- 9744579 TI - Teleradiology: another revolution in radiology? AB - Teleradiology systems are rapidly being deployed by an increasing number of radiological services. Many articles have already been published on the technological developments of teleradiology but little attention has been given to its expected impact on the delivery of health care. This review article will therefore outline the historical and current technological developments of teleradiology and its potential future implementation into mainstream radiology. PMID- 9744580 TI - Chest imaging in AIDS. AB - A review of imaging in the acquired immune deficiency syndrome (AIDS) is presented. The imaging features can be conveniently categorized according to whether the presenting complications are infective (bacterial, protozoal, or fungal), bronchiectasis, neoplastic (Kaposi's sarcoma, AIDS-related lymphoma, or lymphoproliferative disease), or a miscellaneous group (non-specific interstitial pneumonitis, persistent generalized lymphadenopathy, or bronchogenic carcinoma). PMID- 9744581 TI - Pulmonary complications in AIDS: CT appearances. AB - Pulmonary complications are a major cause of morbidity and mortality in patients with AIDS. The aim of this pictorial essay is to familiarize radiologists with the CT appearances of the pulmonary complications of AIDS. These include a wide range of opportunistic infections, neoplastic conditions and lymphoproliferative disorders. Although this is by no means an exhaustive review of the subject, the conditions illustrated constitute the vast majority of pulmonary complications encountered in this setting. PMID- 9744582 TI - Evaluation of the anterior cruciate ligament of the knee: comparison between partial flexion true sagittal and extension sagittal oblique positions during MR imaging. AB - OBJECTIVE: To compare partial flexion true sagittal (FS) magnetic resonance (MR) images with extension sagittal oblique (ESO) MR images with regard to delineation of the anterior cruciate ligament (ACL) in the knee. DESIGN: To establish the appropriate degree of flexion of the knee joint, two human cadaveric knee joints were used as a supplementary technique. FS and ESO images then were performed in 17 knees with an intact ACL and six knees with a torn ACL. In 22 of the 23 knees in which the MR diagnosis of intact or torn ACL corresponded to that derived from arthroscopy, the paired MR images were rated by a three-point scale. RESULTS: FS images were rated superior to ESO images in 53%, 41% and 47% of cases with regard to femoral attachment sites, midportions and tibial attachment sites of intact ACLs, respectively. FS images allowed better assessment of disrupted ACLs and residual ligamentous structures. Overall the FS images were either equal to or better than the ESO in the majority of cases. CONCLUSION: FS images are useful when the ACL is not well visualized in initial ESO images. PMID- 9744583 TI - Complete MR angiography and Doppler ultrasound as the sole imaging modalities prior to carotid endarterectomy. AB - OBJECTIVE: To assess the combination of duplex Doppler ultrasound (DUS) and complete carotid magnetic resonance angiography (MRA) for the non-invasive imaging of carotid disease and their effect on outcomes. Determine inter-reader agreement of carotid MRA. MATERIALS AND METHODS: One-hundred and ten carotid bifurcations were evaluated using DUS, 2D and 3D time-of-flight MRA from the aortic arch to the Circle of Willis in 55 patients. Percentage stenoses were determined by two blinded readers using standardized criteria. Clinical follow-up was by chart review. RESULTS: Correlation of Doppler and MRA was excellent (r=0.903, P<0.001). Inter-reader agreement (K) for MRA was good: internal carotid artery (ICA) (0.750), external carotid artery (ECA) (0.674) and common carotid artery (CCA) (0.410). Differences in CCA readings were due to minor differences in categorizing lesions as CCA versus ICA or ECA. MRA and Doppler detected nine occluded ICAs. Two DUS occlusions had ICA flow by MRA; one due to a reconstituted precavernous ICA, one a near occluded vessel. Five patients (9%) had surgical management modified by MRA with four not having surgery: three distal ICA/Siphon occlusions and one less severe stenosis by MRA. One tandem lesion not visualized by DUS was surgically significant. Nine aortic arch abnormalities had no surgical impact, possibly due to small sample size. Of 41 endarterectomies, there were no complications from errors of diagnosis. CONCLUSION: Carotid MRA correlates well with DUS with good inter-reader agreement. MRA confirms Doppler findings, expands anatomical information and identifies tandem lesions from the aortic arch to the Circle of Willis which can affect surgical management. This approach to carotid artery imaging appears to have no negative effect on surgical outcome. PMID- 9744585 TI - A study on the patency of the inferior mesenteric and lumbar arteries in the incidence of endoleak following endovascular repair of infra-renal aortic aneurysms. AB - OBJECTIVE: An endoleak is defined as the presence of contrast medium within the aneurysm sac on post-operative contrast-enhanced computed tomography scans (CT) in patients following endovascular repair (EVR) of abdominal aortic aneurysms (AAA). The aim of this study was to correlate the incidence of endoleaks with the presence of patent lumbar (LA) and inferior mesenteric arteries (IMA) as seen on pre-operative angiography. DESIGN, MATERIALS AND METHODS: Forty-seven patients were assessed pre-operatively by both CT and angiography by a blinded radiologist prior to EVR of AAA. The number and size of patent vessels was recorded and correlated with the incidence of LA or IMA endoleaks on follow-up CT. Patent lumbar vessels were scored: 1 = small, 2 = medium, 3 = large. RESULTS: Five patients were noted to have patent IMA on pre-operative angiography but none developed an endoleak. In this series, five patients had an endoleak due to a patent LA. The median score for patients with no endoleak was 1 (0-9) and for those with a lumbar endoleak 2 (0-5) (P = 0.26, Mann-Whitney U-test). The number of patent lumbar arteries was not predictive of a subsequent endoleak. Two out of nine (22 %) patients with large patent LA subsequently developed an endoleak. If a policy of pre-operative embolization on the basis of large patent LA had been adopted, seven patients would have had an unnecessary invasive procedure. CONCLUSION: Pre-operative angiography to look for patent LA and IMAs is not required in patients undergoing EVR or AAA. PMID- 9744584 TI - The 'penumbra sign' on T1-weighted MR imaging in subacute osteomyelitis: frequency, cause and significance. AB - OBJECTIVE: We studied the frequency and cause of a feature exhibited on T1 weighted (T1-W) magnetic resonance (MR) imaging termed the 'penumbra sign' in a series of patients presenting with osteomyelitis and correlated it with the double-line sign described as a T2-weighted (T2-W) or short tau inversion recovery (STIR) feature of both the Brodie's abscess and avascular necrosis. MATERIALS AND METHODS: The clinical, radiographic, MR imaging, microbiological and histological findings in 32 patients referred to an orthopaedic oncology service, but subsequently proven to have osteomyelitis, were reviewed. The presence or absence of a rim of tissue lining an abscess cavity typified by minor signal hyperintensity relative to the main abscess contents on T1-W MR imaging (the 'penumbra sign') was identified. The sign was correlated with the radiographic and other findings. RESULTS: The penumbra sign was identified in 24 cases (75%) and appears to be a more sensitive sign than the corresponding double line sign which was evident in only 29% of these on T2-W or fast STIR images. The lesions were unilocular in 11 cases (46%) and multilocular in 13 (54%). The thickness of the penumbra ranged from 2 to 5mm. On histological examination the tissue comprising the penumbra sign was found to be highly vascularized granulation tissue containing thick walled arterioles. CONCLUSION: The penumbra sign is characteristically seen on T1-W MR images in subacute osteomyelitis and is due to a thick layer of highly vascularized granulation tissue which may not be visible as the double-line sign on T2-W or fast STIR sequences. This characteristic, but not pathognomonic, MR finding supports the diagnosis of bone infection and helps to exclude the presence of a tumour. PMID- 9744586 TI - Technical report: The introduction of a new synchronized oesophageal manometry and digital video-fluoroscopy (fluoromanometry) system into the radiology suite. AB - Following the development of a new fluoromanometry system enabling synchronous oesophageal manometry and barium swallow video-fluoroscopy, both the equipment and examination method have been successfully introduced into the radiology suite. The application of the system which uses a PC with video capture and a portable manometry recorder is described together with details of its implementation and the examination technique used. PMID- 9744587 TI - Assessment of the role and reliability of sonographic post-prandial flow response in grading Crohn's disease activity. AB - PURPOSE: To investigate the value of pre and post prandial Duplex colour Doppler sonographic (DCDS) measurement of superior mesenteric artery (SMA) flow in the assessment of Crohn's disease activity, and its response to treatment. MATERIALS AND METHODS: SMA volume flow rates before and after a food challenge (200 ml of Ensure Plus) were recorded over 60 min in 11 controls, and 25 patients with proven Crohn's disease. Peak flow rates and the time interval to peak flow were recorded. Eleven patients with active disease were monitored longitudinally and their response following the introduction of systemic steroids was assessed. RESULTS: The time interval from food challenge to peak SMA flow rate was significantly lower in patients with untreated active disease (median 20 min, range 14.5-21.25) compared to inactive patients (median 33 mins, range 28.75 40.5, P = 0.0006). Longitudinal follow-up of active disease demonstrated prolongation of time to peak flow following clinical remission (P = 0.0024) CONCLUSIONS: This technique is useful in offering an immediate, noninvasive means of assessing disease activity. Further longitudinal follow up data is necessary to determine its utility in assessing response to treatment. PMID- 9744588 TI - Magnetic resonance imaging features of pituitary yttrium-90 rod implantation. AB - Intersellar implantation of yttrium-90 rods was a common treatment for a variety of pituitary tumours in the 1960s and 1970s. The magnetic resonance (MR) imaging features in three patients with implants (two for growth hormone-secreting and one for prolactin-secreting pituitary adenomas) are presented: the implants appeared as low signal cylinders with no image distortion, in contradistinction to CT where the implants generate beam hardening and back projection artefacts. Confident evaluation of the pituitary fossa for residential tumour and sequelae of therapy could be made on MR. It is the imaging technique of choice in the follow-up of patients treated with yttrium-90 implants. PMID- 9744589 TI - Result of angioplasty of brescia-cimino haemodialysis fistulae: medium-term follow-up. AB - AIM: To determine the results of transluminal angioplasty in patients with Brescia-Cimino arteriovenous fistulae. PATIENTS AND METHODS: Thirty-one patients underwent transluminal angioplasty of 36 stenotic lesions related to Brescia Cimino arteriovenous fistulae over a 5-year period. The lesions treated were characterized by review of pre-angioplasty fistulograms. Medical and radiological records were reviewed to assess medium-term patency of each patient's fistula. RESULTS: Angioplasty was performed successfully in 28 out of 31 patients initially (90% technical success rate). Duration of follow-up for the 31 patients ranged from 4 to 65 months (median = 34 months). At 6 months, seven patients required further surgical or endovascular intervention (18 patients remained event-free) and at 1 year, 10 patients required further endovascular or surgical intervention (14 patients remained event-free). Life-table analysis revealed primary patency rates of 77%, 64% and 39% at 6 months, 1 year and 2 years, respectively. At 6 months and 1 year, four and five patients, respectively, required surgical revision or closure of fistula. Secondary patency rates were 85%, 81% and 65% at 6 months, 1 year and 2 years, respectively. All patients with a primary patency at 2 years remained event-free during the follow-up period. CONCLUSIONS: Transluminal angioplasty is an effective treatment for stenoses developed in relation to Brescia-Cimino haemodialysis fistulae. Further endovascular procedures may be required, especially in the first 24 months, to preserve patency. These techniques extend the lifetime of fistulae, thereby preserving proximal venous access sites for future use. Our result is in broad agreement with results from other series. PMID- 9744591 TI - Case report: Expiratory helical CT scan minimum intensity projection imaging in cystic fibrosis. PMID- 9744590 TI - A comparison of Fleet Phospho-soda with Picolax in the preparation of the colon for double contrast barium enema. AB - This study was performed to compare the patient acceptability and the efficacy of two different agents for bowel preparation prior to double contrast barium enema. One-hundred and ninety-four outpatients were randomized to have either two sachets of Picolax or two bottles of Fleet Phospho-soda and restricted to clear fluids on the day prior to their examination. Patients answered a short questionnaire before their enema. The decubitus films were assessed for faecal residue and bowel coating by three observers blinded to the type of preparation used. There was no significant difference in faecal residue nor in the bowel coating between the preparations. However patients found Picolax significantly easier to take, being better tasting and provoking less nausea and vomiting than Fleet Phospho-soda. PMID- 9744593 TI - Case report: Femoral hernia causing small bowel obstruction--ultrasound diagnosis. PMID- 9744592 TI - Case report: The treatment of a chronic parotid cutaneous fistula by the injection of a solution of lipiodol with cyanoacrylate. PMID- 9744594 TI - Case report: Meningioma presenting as an aural polyp. PMID- 9744595 TI - Grey-scale ultrasound appearances of chronic parotid enlargement in anorexia nervosa. PMID- 9744596 TI - Primary non-Hodgkin's splenic lymphoma. PMID- 9744598 TI - Safety and efficacy of pancreatic sphincterotomy in chronic pancreatitis. AB - BACKGROUND: Endoscopic pancreatic sphincterotomy (EPS) is being performed with increasing frequency as a prerequisite to interventional measures in the pancreatic duct. The aim of this study was to evaluate EPS with regard to technique, success, complications, and mortality in patients with chronic pancreatitis. METHODS: Between January 1989 and September 1996, the results of all consecutive EPSs in patients with chronic pancreatitis were documented in a standardized form. Patients were followed by clinical investigation and blood sample analysis at 4, 24, and 48 hours after EPS. Complications were classified according to commonly accepted criteria. RESULTS: EPS was performed in 118 patients with chronic pancreatitis (men 75%, women 25%, 48+/-10 years). Ninety four patients (80%) underwent guidewire-assisted EPS, and 24 patients (20%) underwent needle-knife EPS. Seventy-seven EPS procedures (65%) were primarily successful (guidewire EPS: 60 of 94, 64%; needle-knife EPS: 17 of 24, 71%). Additional endoscopic cutting techniques (needle-knife papillotomy, biliary endoscopic sphincterotomy) were required in 41 patients (35%). In total, EPS was successful in 116 patients (98%). The complication rate was 4.2% (4 cases of moderate pancreatitis, 1 severe bleeding, no deaths). All complications were managed nonoperatively. CONCLUSIONS: In patients with chronic pancreatitis, EPS with a standard sphincterotome or with a needle-knife offers an effective and reliable approach to the pancreatic duct system. Additional cutting techniques may be necessary in approximately one third of cases before an EPS can be successfully performed. The complication rate of EPS in patients with chronic pancreatitis appears to be lower than the complication rate of biliary sphincterotomy for other indications. PMID- 9744597 TI - Seroprevalence of immunoglobulin G antibodies against Helicobacter pylori among endoscopy personnel in Japan. AB - BACKGROUND: The seroprevalence of immunoglobulin G antibodies against Helicobacter pylori in endoscopy personnel was determined to investigate whether gastrointestinal endoscopists and endoscopy nurses in Japan are at an increased risk for H. pylori infection and to clarify risk factors for H. pylori infection during endoscopy. METHODS: One hundred twenty-one gastrointestinal endoscopists and endoscopy nurses provided personal information, including their observance of infection-control measures, by means of self-administered questionnaire. One hundred one age-matched healthy individuals undergoing routine physical examinations served as controls. Serum samples from each subject were examined with enzyme-linked immunosorbent assay for the presence of IgG antibodies against H. pylori. RESULTS: Among younger subjects (< 40 years old), endoscopists and endoscopy nurses had higher seropositive rates than did control subjects (p < 0.05). Among older subjects (> or = 40 years old), the seropositive rate did not differ between endoscopy personnel and control subjects (p = 0.2174). However, among older seropositive subjects, endoscopy personnel had significantly higher antibody titers than did control subjects (p < 0.01). Older seropositive endoscopists performed significantly more examinations per month than did their seronegative colleagues (p < 0.05). Furthermore, younger seropositive endoscopy nurses performed significantly more examinations per month than did seronegative nurses (p < 0.05). CONCLUSIONS: Gastrointestinal endoscopists and endoscopy nurses in Japan are at high risk for H. pylori infection. The risk of H. pylori infection is correlated with the frequency of endoscopic examinations, especially in older gastrointestinal endoscopists and younger endoscopy nurses. PMID- 9744599 TI - Is there a synergistic effect between mixed bacterial infection in biofilm formation on biliary stents? AB - BACKGROUND: Biliary sludge which forms as a result of bacterial adherence and biofilm formation in the biliary system is a recognized cause of blockage of plastic stents. Bacteriological cultures of sludge have revealed a mixed infection with gram-positive and gram-negative bacteria. Animal studies have shown that prophylactic ciprofloxacin, which selectively suppress gram-negative bacteria, results in prolonged stent patency despite colonization of the stents by gram-positive bacteria. METHODS: We tested a possible synergistic effect between gram-negative and gram-positive bacteria in adherence and biofilm formation on plastic stents. Clinical isolates of Escherichia coli and Enterococcus were cultured in separate chemostats to achieve a steady growth. Adherence of the two bacteria on plastic stent surface were tested separately by perfusing infected bile with the respective bacteria through different modified Robbins devices containing 10F polyethylene stent pieces up to 4 days. In a second experiment, Enterococcus was perfused through stent pieces precolonized with E. coli for 24 hours. The stent pieces were then removed daily and analyzed by bacteriologic culture and scanning electron microscopy for bacterial adherence and biofilm formation. RESULTS: Gram-negative E. coli were more adherent than gram-positive Enterococcus. Precolonization with E. coli facilitates subsequent attachment of Enterococcus. CONCLUSIONS: We concluded that there is a synergistic effect between gram-positive and gram-negative bacteria in adherence and biofilm formation. PMID- 9744600 TI - Ballooning of the papilla during contrast injection: the semaphore of a choledochocele. AB - BACKGROUND: Choledochocele is a rare abnormality involving the intramural segment of the common bile duct. ERCP is essential to demonstrate a choledochocele. The aim of this study was to investigate the duodenoscopic and cholangiographic findings during ERCP. METHODS: Over a 4-year period, 17 symptomatic patients (8 men, 9 women; age range 45 to 83 years) were identified as having choledochoceles. The diagnosis of choledochocele was made by both duodenoscopic and cholangiographic findings. RESULTS: As a characteristic duodenoscopic finding, the enlarged bulging papilla was noted in 8 patients (47%), whereas a normal-appearing papilla was noted in 9 patients (53%) before the injection of contrast media. However, in all 17 patients progressive enlargement or ballooning of the papilla was noted during contrast injection. The maximum diameter of the choledochocele, determined by cholangiography, was significantly larger (19 +/-4 mm) in patients with initially bulging papilla than in those with normal appearing papilla (9+/-3 mm) (p < 0.05). CONCLUSIONS: Ballooning of the papilla during contrast injection may be a specific duodenoscopic finding for choledochocele. To avoid missing the diagnosis of a choledochocele, especially when it is small, it is important to watch the papilla carefully during contrast injection. PMID- 9744601 TI - Marginal irregularity of flat elevated type of colorectal tumor as a marker of malignant potential. AB - BACKGROUND: The malignant potential of superficial elevated type tumors is still controversial. Because biopsy specimens alone are sometimes not sufficient for diagnosis due to histologic heterogeneity within the tumor, other clinical parameters for evaluation of the degree of malignancy would be helpful. METHODS: A new morphometric parameter, the "F-circle," which represents the irregularity of the tumor margin, was studied in 115 endoscopically recognized superficial, flat, elevated type colorectal tumors without a central depressed area with respect to histologic evaluation of the degree of malignancy. RESULTS: The mean F circle values of adenomatous lesions with mild or moderate atypia, severe atypia, and adenocarcinoma were 0.709+/-0.115, 0.619+/-0.149, and 0.536+/-0.133, respectively. CONCLUSION: The superficial elevated type of colorectal tumor having more marginal irregularity had a greater malignant potential, and the F circle may be a useful clinical parameter for evaluation of the degree of malignancy. PMID- 9744602 TI - Self-expandable metallic stents in the palliation of rectosigmoidal carcinoma: a follow-up study. AB - BACKGROUND: Currently applied endoscopic palliative treatment of advanced rectosigmoidal carcinoma is hampered by the cost of the equipment, the need for repeated, often painful treatment sessions, and the occurrence of complications. Metallic expandable stents are effective in the palliation of malignant esophageal and biliary stenoses. We evaluated the use of a new type of self expandable nitinol stent in the palliation of rectosigmoidal carcinoma. METHODS: In 10 patients with advanced obstructing rectosigmoidal carcinoma, initial Nd:YAG laser treatment was performed if necessary to allow passage of a gastroscope. Subsequently, a self-expanding nitinol stent with flanged ends was inserted under combined fluoroscopic and endoscopic control. Endoscopic and clinical follow-up was carried out at regular intervals. RESULTS: After 2+/-0.4 sessions of initial laser therapy, minimal lumen diameter was 9+/-1 mm. Stent insertion was successful in 9 patients, increasing minimal lumen diameter to 14+/-1.2 mm (p < 0.005). In one patient, stent deployment was complicated by a sigmoid perforation, requiring surgery. After insertion, colorectal stents remained adequately positioned and free of obstruction for 103+/-31 days. Patient survival after stent placement was 204 +/-43 days. Stent migration occurred in 3 patients, after 38+/-10 days. Obstruction of the stent because of tumor ingrowth was observed in only one patient, after 268 days. CONCLUSION: Insertion of self expandable nitinol stents in patients with rectosigmoidal carcinoma is technically feasible. Metallic stents are effective in the palliation of malignant rectosigmoid obstruction; they provide an alternative to repeated palliative laser therapy or palliative surgery. PMID- 9744603 TI - A new method of evaluating hemorrhoids with the retroflexed fiberoptic colonoscope. AB - BACKGROUND: The conventional classification of the degree of hemorrhoids does not consider the severity of hemorrhage. The purpose of this study was to establish a new objective method for evaluating hemorrhoids in close relation to the main symptoms, hemorrhage and prolapse, as observed through a retroflexed colonoscope in the rectum. METHODS: The subjects were 531 consecutive patients who complained of symptoms related to the rectum or the anus. The degree of mucosal elevation of the rectal columns, changes in color (the existence and degree of red color sign, dilated vein, and white area), and the existence and size of hypertrophied anal papillae were evaluated by colonoscopy. RESULTS: Red color sign was the finding closely related to hemorrhage (p < 0.0001). Dilated vein, white area, and a large hypertrophied anal papilla were related to prolapse (p < 0.0001). The degree of mucosal elevation of the rectal columns was related to both hemorrhage and prolapse (p < 0.0005, p < 0.05). CONCLUSION: Retroflexing the colonoscope intrarectally facilitated identification of findings in the anal canal related to hemorrhage and prolapse, which are the clinical manifestations of hemorrhoids. PMID- 9744604 TI - Meta-analysis and cost comparison of polyethylene glycol lavage versus sodium phosphate for colonoscopy preparation. AB - BACKGROUND: Although polyethylene glycol lavage solutions are widely used for colonoscopy preparation, evidence suggests that sodium phosphate is better tolerated and has similar efficacy. The purpose of this study was to compare compliance with and efficacy of polyethylene glycol and sodium phosphate using meta-analysis and to compare the cost of colonoscopy with both methods. METHODS: We used Medline to identify all randomized controlled trials comparing the two preparations. Study methods were evaluated, and quantitative data were abstracted independently, including inability to complete the preparation and preparation quality, rated as adequate or excellent. A random effects model was used to calculate the pooled relative risk. Direct costs and literature-based probability estimates were used to compare costs. RESULTS: Among 1286 subjects from eight colonoscopist-blinded trials, the pooled relative risk of inability to complete the preparation was 0.23 (95% CI [0.18-0.28]) in favor of sodium phosphate. Although the best estimate of the relative risk for an adequate quality preparation revealed therapeutic equivalence (relative risk = 1.06: 95% CI [0.95 1.19]), an excellent quality preparation was more likely with sodium phosphate (relative risk = 1.72: 95% CI [1.16-2.53]). Assuming reexamination rates from published literature of 3% and 8% for sodium phosphate and polyethylene glycol, respectively, direct costs of colonic examination were $465 and $503. There were no clinically important adverse effects with either method. CONCLUSION: The results suggest that sodium phosphate is as effective and less costly, with a more easily completed preparation, compared with polyethylene glycol and is the preferred method of preparation for colonoscopy for certain patient subgroups. PMID- 9744605 TI - Sublingual hyoscyamine for patient comfort during screening sigmoidoscopy: a randomized, double-blind, placebo-controlled clinical trial. AB - BACKGROUND: Screening sigmoidoscopy is an underutilized method for detecting early colorectal cancer, and patient discomfort is one reason for poor compliance in the general population. The possible benefit of a well-tolerated, low-cost antispasmodic medication, sublingual hyoscyamine, used before flexible sigmoidoscopy was assessed in a randomized, double-blinded, placebo-controlled trial. METHODS: One hundred fifty patients were enrolled and randomized to receive two sublingual hyoscyamine tablets (0.125 mg/tablet) or the placebo 10 minutes before sigmoidoscopy. Patient comfort and the endoscopist's perception of the ease of insertion were measured using a 100 mm visual analog scale. The depth of sigmoidoscope insertion was measured in centimeters, and complications were recorded. RESULTS: The median age was 55 years (range 25 to 83 years). There were 100 men (66.7%) and 50 women (33.3%). Approximately half (n = 76, 50.7%) had a prior sigmoidoscopy or colonoscopy. No statistical differences were found between treatment group means for age, gender, pain score, ease of insertion, or depth of insertion. The hyoscyamine group tended to have lower mean pain (32.4 vs. 37.7, p = 0.18) and difficulty (29.9 vs. 33.7, p = 0.31) scores and greater depth of sigmoidoscope insertion (51.3 vs. 47.7, p = 0.07); however, the differences were not statistically significant. The treatment groups differed with a higher percentage of the hyoscyamine group having a previous endoscopy (60.0% vs. 41.3%, p = 0.02); however, no significant differences were detected between mean pain scores as related to treatment when controlling for previous experience with endoscopy (p = 0.31). CONCLUSIONS: In this study, hyoscyamine administered in the sublingual route did not significantly improve patient comfort, ease of insertion, or the depth of sigmoidoscope insertion during screening sigmoidoscopy. The search for alternative methods to improve patient comfort during screening endoscopy should continue. PMID- 9744606 TI - Fascioliasis causing obstructive jaundice. PMID- 9744607 TI - Endoscopic retrograde wire-guided cytology of malignant biliary strictures using a novel scraping brush. AB - BACKGROUND: Tissue sampling to differentiate benign from malignant pancreatobiliary strictures remains problematic despite the availability of several new sampling methods. A new device is described which attempts to correct some of the drawbacks. METHODS: The device consists of a 10F dilator which has an attached pad of Velcro. The Velcro has semi-rigid, mushroom-shaped bristles. A cytologic sample is obtained by the abrasive action of the brush when it is passed through the stricture. Fifteen patients with obstructive jaundice underwent brushing of the bile duct using this device. RESULTS: Cytologic samples obtained with this device were positive for malignancy in all 15 patients. Diagnostic confirmation was obtained by assessment of clinical course, radiologic findings, and during surgery. CONCLUSION: Preliminary experience indicates that this new device enhances the yield of tissue sampling from malignant bile duct strictures. PMID- 9744608 TI - Tissue injury of Injection Gold Probe. AB - BACKGROUND: The Injection Gold Probe, which incorporates an injection needle and a bipolar electrocoagulation probe, potentially offers better anchoring of the probe for electrocoagulation. This study compares the tissue injury caused by the Injection Gold Probe with or without protrusion of the needle. METHODS: A 10F Injection Gold Probe was applied perpendicularly on the antrum of porcine stomach. At each site of testing, one pulse of treatment at 12 W was given for 10 seconds. The extent of tissue injury was studied under different forces of application (light touch or maximal force) and different probe conditions (needle in, needle-out, needle-out plus 1 mL saline solution injection). Each maneuver was repeated 8 times. The depth and width of tissue injury were assessed histologically by a pathologist blinded to the treatment. RESULTS: The depth and width of tissue injury did not differ significantly with needle-in, needle-out, or needle-out plus 1 mL saline solution injection on both light touch. But the tissue injury was significantly deeper and wider with maximal force applications than with light touch under the same probe condition. CONCLUSION: The extent of tissue injury inflicted by electrocoagulation using the Injection Gold Probe is not affected by the position of the injection needle. PMID- 9744609 TI - Retrograde approach to pharyngo-esophageal obstruction. AB - BACKGROUND: The entrance of the esophagus has to be identified for treatment of a pharyngo-esophageal obstruction. If transoropharyngeal identification is unsuccessful, a retrograde approach might be indicated. METHODS: By way of a mini laparotomy and gastrotomy, a flexible gastroscope can be passed into the esophagus. In one patient with a Zenker's diverticulum, a guidewire was inserted through the accessory channel of the gastroscope and passed through a stenosis, caused by marked hypertrophy of the cricopharyngeal muscle, into the oral cavity. Thereafter antegrade dilatation and laser assisted myotomy could be performed. In another patient with a membranous obstruction of the esophageal entrance due to radiotherapy, the occlusion was perforated transoropharyngeally and bluntly dilatated guided by the light from the gastroscope. RESULTS: In both cases the esophageal passage was restored. No complications occurred as a result of the procedures. CONCLUSIONS: The retrograde approach may be a good alternative when antegrade identification of the esophageal entrance fails. PMID- 9744610 TI - Gastrointestinal hemorrhage consequent to foreign body reaction to silk sutures: case series and review. PMID- 9744611 TI - Endoscopic management of rectal Dieulafoy-like lesions: a case series and review of literature. PMID- 9744612 TI - Enteroliths in a Kock continent ileostomy: endoscopic diagnosis and management. PMID- 9744613 TI - Endoscopic transgastric drainage of a retroperitoneal lymphocele. PMID- 9744614 TI - Disseminated peritoneal adenomucinosis--a diagnostic approach by peritoneoscopy. PMID- 9744615 TI - Double-channel endoscopic polypectomy technique for the removal of large pedunculated polyps. PMID- 9744616 TI - Endoscopy personnel and the transmission of Helicobacter pylori infection. PMID- 9744617 TI - Digital imaging in endoscopy. PMID- 9744618 TI - Helicobacter pylori infection in patients with early gastric cancer by the endoscopic phenol red test. PMID- 9744619 TI - Randomised trial of octreotide for long term management of cirrhosis after variceal hemorrhage. PMID- 9744620 TI - Endoscopic management of Dieulafoy lesions of the stomach: a case study of 26 patients. PMID- 9744621 TI - Laparoscopic cholecystectomy and ERCP--lessons from the Danish National Registry. PMID- 9744622 TI - Angiographic and endoscopic management of bleeding pancreatic pseudo-aneurysms. PMID- 9744623 TI - Endoscopic clipping of duodenal Dieulafoy's lesions: alone or combined? PMID- 9744624 TI - Colonic cavernous hemangioma. PMID- 9744625 TI - Colonoscopic clipping closure of a divot after the removal of an impacted chicken bone. PMID- 9744626 TI - Proton spectroscopy in children with epilepsy and febrile convulsions. AB - An association between complex febrile convulsions and the development of hippocampal atrophy, which is characterized by neuron loss and gliosis, has been suggested but is still controversial. In proton magnetic resonance spectroscopy (1H-MRS) a reduction in N-acetylaspartate (NAA), a neuron marker, or in its ratio to other metabolites, that is, creatine and phospocreatine (Cr) and choline containing compounds (Cho), is considered a sensitive method for detecting neuron loss. We performed 1H-MRS of mesial temporal regions, including hippocampi, in two different groups of children with epilepsy: in children with a history of complex febrile convulsions (CFCs) (n = 7; mean age 7.1 years) and in children without any history of CFCs, referred to herein as the non-CFC group (n = 6; mean age 7.6 years). Changes in the metabolite ratios were detected in 57% of children in the CFC group and in 67% of children in the non-CFC group. In both groups, NAA/(Cho + Cr), NAA/Cho, and NAA/Cr were significantly decreased ipsilaterally to the seizure focus when compared with the control group, but no significant differences were detected between the CFC and non-CFC groups. Also on the contralateral side, NAA/(Cho + Cr) and NAA/Cr were significantly decreased in both patient groups, but the differences were not significant between the CFC and non-CFC groups. Metabolite abnormalities in the mesial temporal region were detected in children with intractable epilepsy and in children whose epilepsy is well controlled by antiepileptic medication. The noninvasive 1H-MRS can be considered an additional diagnostic method to promote early detection of mesial temporal abnormalities that, in the light of this study, seem to be underdiagnosed in children with either temporal lobe epilepsy or other seizure types. PMID- 9744627 TI - Intracranial arachnoid cysts in children: related signs and associated anomalies. AB - Intracranial arachnoid cysts are benign development anomalies that may be clinically asymptomatic. The authors describe 30 children with intracranial arachnoid cysts in terms of clinical manifestations and relations to the associated brain anomalies or lesions. The mean age at onset of clinical manifestations was 4 years, 7 months (range 1 day to 14 years). The mean age at diagnosis was 6 years, 2 months (range 10 days to 16 years). Most patients with nonprogressive symptoms, such as seizures and headache, had focal epileptiform discharges on electroencephalogram, and they benefited from antiepileptic drugs. Surgery resulted in only partial reduction in both cyst size and seizure frequency in patients with intractable seizures, and it also failed to improve some neurologic signs, such as sexual precocity or cranial neuropathy resulting from long-term compression of arachnoid cysts. We conclude that the only absolute indication for surgery is the presence of progressive hydrocephalus or intracranial hypertension. The associated anomalies or lesions include brain tumors, giant nevocellular nevi, achondroplasia, microphthalmia, intracystic hemorrhage, dysgenesis of the corpus callosum, and heterotopia. PMID- 9744628 TI - Cerebral perfusion in children with Alice in Wonderland syndrome. AB - Alice in Wonderland syndrome (AIWS) is characterized by visual hallucinations and bizarre perceptual distortions. Technetium-99m hexamethylpropyleneamine tomography (SPECT) brain scans were performed in four patients during the acute stage of AIWS. Two patients were demonstrated to have Epstein-Barr virus infections. One had abnormal (EEG) findings. The visual-evoked potential, cranial CT, and MRI findings were negative. The decreased cerebral perfusion areas in all patients were near the visual tract and visual cortex. All involved some regions of the temporal lobe. In most patients with AIWS, the EEG, CT, and MRI are unable to determine the precise pathologic areas. However, a SPECT brain scan may demonstrate abnormal perfusion areas and explain the clinical presentations. PMID- 9744629 TI - Brainstem auditory-evoked potential evaluation in children with meningitis. AB - Brainstem auditory-evoked potential (BAEP) was performed on 101 children with meningitis to assess the incidence of hearing impairment. Fifty-two (51.5%) children had bacterial meningitis, six (5.9%) had viral meningitis, and 43 (42.6%) had aseptic meningitis. Fifty-one (50.5%) patients were assessed before discharge and 50 (49.5%) 9 days to 17 months later (mean = 4 months). BAEP impairment was found in 28 (27.7%) of 101 patients; 24 had sensorineural and four had conductive type of hearing loss, and 17 (60.7%) had unilateral and 11 (39.3%) had bilateral impairment. Hearing threshold was elevated in 22 (21.8%) patients, and the other six had increased latency and interpeak latencies with normal threshold. Frequency of BAEP impairment or hearing loss associated with bacterial meningitis was 34.6% and 30.8%, respectively; frequency associated with aseptic meningitis was 20.9% and 13.9%, respectively. One child with viral meningitis (coxsackie virus) had mild BAEP impairment. Most of the BAEP impairment in the bacterial meningitis group was associated with H. influenzae. Prospective BAEP study was performed in 20 patients randomly at 0.3 to 18 months to assess hearing status after antibiotic treatment, 10 with normal and 10 with abnormal BAEP. All the initially normal BAEP patients remained normal. Of the 10 patients with abnormal BAEP results initially, four returned to normal, two improved, three remained unchanged, and one deteriorated. The incidence of hearing loss after bacterial and aseptic meningitis is high. BAEP is useful to screen for possible hearing loss in children with meningitis, and follow-up BAEP is necessary for those patients with initially abnormal BAEP. PMID- 9744630 TI - Choreoathetosis after cardiac surgery with hypothermia and extracorporeal circulation. AB - Eleven children, 4-48 months old, with congenital cyanotic heart defects developed choreoathetoid movements 2-12 days after cardiac surgery with hypothermia and extracorporeal circulation (ECC). The abnormal movements mainly involved the limbs, facial musculature, and tongue, leading to a severe dysphagia. The symptoms had an acute onset, after a period of apparent neurologic normality, and had a variable outcome. Of the nine children that survive, three had abnormal movements when last seen (41 days to 12 months of follow-up). The other six children had a complete regression of the choreoathetoid movements 1-4 weeks after onset. No specific finding was observed in the CT scans, cerebrospinal fluid examination, or EEG that could be related to the abnormal movements. Symptomatic therapy with haloperidol with or without benzodiazepines led to symptomatic improvement in six children, although there was no evidence that this treatment modified the evolution of the disease. The authors conclude that the choreoathetoid syndrome after cardiac surgery with deep hypothermia and ECC is an ill-defined entity requiring additional study to better understand its pathogenesis so that preventive measures can be taken to avoid a condition that can lead to permanent and incapacitating neurologic sequelae. PMID- 9744631 TI - Cerebral complications of nonaccidental head injury in childhood. AB - Patterns of cerebral parenchymal injury and their relationship to outcome morbidity are evaluated in this retrospective study of 14 children with confirmed nonaccidental head injury (NAHI). The mean age at time of injury was 12 months 6 days, mean Children's Coma Score was 5.36, and mean postinjury follow-up was 17 months 12 days. All patients had acute subdural hematoma (interhemispheric or convexity) on initial CT imaging. Two major groups of children were identified from initial CT scans; those with diffuse cerebral hypoattenuation (n = 7) and those with focal cerebral hypoattenuation (n = 7). The two groups differed significantly by age (diffuse group, mean age 5 months 9 days +/- 36 days; focal group, mean age 19 months 3 days +/- 6 months 9 days; P < 0.01) and ultimate type and extent of parenchymal damage. Outcome was generally poor in both groups (mean Children's Outcome Score of III/IV). Cerebral infarction developed in all survivors. Most common were hemispheric necrosis after hemispheric swelling subjacent to an ipsilateral convexity acute subdural hematoma (n = 5); distribution of the posterior cerebral artery (n = 4) or callosomarginal branch of the anterior cerebral artery (n = 4); and borderzone infarctions (n = 4). Of 14 children, 11 (79%) had early posttraumatic seizures (EPTS). Clinical progression of symptoms was confirmed in nine patients (mean Childrens Coma Score was 4.0 +/- 0.33). None had a lucid interval. This is the first study using strict inclusion criteria that documents the range of infarction patterns and potential age-dependent differences in postinjury response cascades after nonaccidental head injury. PMID- 9744632 TI - Evaluation of bone mineral density in children receiving antiepileptic drugs. AB - The effects of the valproic acid and carbamazepine monotherapies on bone mineral density were evaluated. Bone mineral density was measured in 53 children with primary epilepsy taking either valproic acid (n = 25) or carbamazepine (n = 28) for longer than 1 year and in a healthy control group (n = 26) by the dual-energy x-ray absorptiometry method at L2-L4 levels of lumbar vertebrae. The mean serum levels of valproic acid and carbamazepine were 66 +/- 2.2 microg/mL and 7.0 +/- 9.3 microg/mL, respectively, and the mean duration of treatment for each drug was 2.4 +/- 0.2 years and 2.6 +/- 0.5 years, respectively. Calcium intakes in diet were similar in both the control and study groups. The serum levels of calcium and phosphorus in all groups were normal. Bone mineral density values of both valproic acid and carbamazepine groups were not statistically different from that of the control group (P > 0.05). PMID- 9744633 TI - Similar brain SPECT findings in subclinical and clinical seizures in two neonates with hemimegalencephaly. AB - Brain single-photon emission computed tomography (SPECT) findings during clinical and subclinical seizures were compared in two neonates with hemimegalencephaly. Interictal and ictal brain SPECT were performed in two neonates. The ictal studies were performed during a clinical seizure in one neonate and during a subclinical seizure in another neonate. They revealed similar focal hemispheric hyperperfusion at the electroencephalographic seizure foci in both cases. The similar perfusion patterns imply that clinical and subclinical seizures place similar metabolic demands on the cerebral tissue involved in the generation of electroencephalographic seizures in neonates with cerebral dysgenesis and suggest that clinical and subclinical seizures should be treated similarly in this population. PMID- 9744635 TI - MRI findings in a neonate with cerebellar agenesis. AB - Cerebellar agenesis is a rarely observed malformation that is frequently associated with other defects. We describe a neonate with an isolated cerebellar agenesis. In addition to the absence of recognizable cerebellar tissue, cranial magnetic resonance imaging demonstrated a hypoplastic base of the pons and absence of the normal outline of the inferior olives. Other major cerebral malformations were not found. As a developmental defect, cerebellar agenesis is heterogeneous because it occurs either as an anatomically isolated anomaly or as part of a more complex cerebral malformation. The pathogenesis and molecular basis of isolated cerebellar agenesis is unknown. PMID- 9744634 TI - Myelopathy resulting from invasive aspergillosis. AB - Aspergillus, a ubiquitous mold, may cause invasive and fatal disease in immunosuppressed patients. Myelopathy is an uncommon presentation of invasive aspergillosis. This report describes three children admitted to the hospital between 1988 and 1995 who developed myelopathy as the first evidence of invasive aspergillosis. All had advanced leukemia and were profoundly immunosuppressed because of chemotherapy and broad-spectrum antibiotics. Weakness and pain presented first; then, sensation to pain and temperature was lost 2 to 6 days later, followed by complete myelopathy. Multiple brain lesions were seen on magnetic resonance imaging in one patient. Despite antifungal therapy, aspergillosis proved fatal within 1 month of onset of myelopathy in all patients. Physicians caring for immunocompromised children should be aware of myelopathy as a presentation of invasive aspergillosis. PMID- 9744636 TI - Valproate therapy: predisposition to bone fracture? AB - A 13-year-old girl on valproate therapy had 20 fractures over a 4-year period between the ages of 5 years and 9 years. Once valproate was withdrawn, no further fractures occurred over the ensuing 4 years. Three other children manifested at least two fractures while on valproate antiepileptic therapy. These reports suggest that valproate, along with other known causes of demineralization (e.g., lack of exercise, diet, and genetic factors), predisposes patients to fractures. PMID- 9744637 TI - Brainstem lesion in Aicardi-Goutieres syndrome. AB - Aicardi-Goutieres syndrome is characterized by the calcification of basal ganglia, leukodystrophy, and lymphocytosis in cerebrospinal fluid. No brainstem lesion has been described. We report a Japanese girl who presented with delayed development and microcephalus at early infancy. Magnetic resonance imaging revealed T2-weighted high intensity in the cerebral white matter and brainstem, and nerve conduction velocity was delayed in the central nervous system, indicating that she manifested dysmyelination in the brainstem white matter similar to that in the cerebral white matter. PMID- 9744638 TI - Drop episodes in Coffin-Lowry syndrome: exaggerated startle responses treated with clonazepam. AB - A 16-year-old girl had fully manifested Coffin-Lowry syndrome and drop episodes. Her drop episodes were precipitated by sudden unexpected tactile or auditory stimuli associated with the electrostatic circumstances in her leg muscles immediately after the stimuli. Studies revealed that her drop episode symptom was an unusual type of startle response and that it may be associated with Coffin Lowry syndrome. PMID- 9744639 TI - MRI nerve root enhancement in Krabbe disease. AB - Krabbe disease is characterized by abnormal breakdown and turnover of myelin, leading to extensive demyelination in both the peripheral and central nervous systems. A 7-month-old infant with early-onset Krabbe disease had deceptively normal head images, but spinal MRI demonstrated abnormal gadolinium enhancement of the lumbosacral sacral nerve roots and cauda equina such as that seen in Guillain-Barre syndrome. Abnormal enhancement in spinal MRI has not been previously described in patients with leukodystrophies. PMID- 9744640 TI - Pseudotumor cerebri as a presenting symptom of acute sinusitis in a child. AB - Pseudotumor cerebri is a clinical syndrome characterized by increased intracranial pressure in the absence of an intracranial tumor. It is most frequently diagnosed in obese young women, but it is also reported in children of all age groups, including infants. A variety of medical conditions have been suggested as possible etiologic factors, including several infectious diseases. This study presents a child with pseudotumor cerebri as the only presenting symptom of acute frontal sinusitis. The possible association between these two conditions should be investigated in cases of pseudotumor cerebri to enable the appropriate treatment. PMID- 9744641 TI - Respiratory failure in nemaline myopathy. PMID- 9744642 TI - Anticatabolic and anabolic strategies in critical illness: a review of current treatment modalities. AB - Critically ill patients characteristically exhibit a pronounced catabolism in addition to a down-regulation of normal anabolic activity, leading to major complications from loss of body protein stores. The marked decrease in lean body mass and protein stores leads to the loss of essential structural and functional proteins required for restoring and maintaining homeostasis. The standard management of the catabolic response to injury and illness has centered on optimizing nutrient intake that modulates but does not reverse the process. Complications of ongoing catabolism therefore remain a major cause of morbidity. Addition of anticatabolic and anabolic agents that may counteract "the stress response to injury or illness" may be of significant clinical benefit. Agents currently available for clinical use, which will be described, can be divided into two groups. The first group are nutrients and nutrient metabolites, namely protein and the specific amino acids, glutamine, arginine, and branched chain amino acids, especially leucine. The second group are anabolic hormones, namely growth hormone, testosterone, and the testosterone analog oxandrolone. The pros and cons of these agents, as to their anabolic and anticatabolic value, are described. PMID- 9744644 TI - The physiologic consequences of macrophage pacification during severe acute pancreatitis. AB - Macrophage overproduction of inflammatory mediators is detrimental in the progression of acute pancreatitis. Although inhibition of inflammatory mediators has been shown to decrease the severity of experimental pancreatitis and improve overall survival, less is known about the mechanism by which blockade produces these benefits. Prior to the induction of lethal acute pancreatitis, rats were randomized to receive a single dose (.01, .1, 1.0, or 10 mg/kg) of a macrophage pacifying compound (CNI-1493) or vehicle. Escalating doses provided incremental increases in survival from 10% (vehicle) to a maximum of 70% (CNI-1493, 1.0 mg/kg). To evaluate the physiologic mechanism responsible for the improved survival, continuous arterial blood pressure, serial hematocrit, ascites volume, pancreatic edema, bronchoalveolar leukocytes and protein, and pancreatic histology were determined in additional rats receiving CNI-1493 (1.0 mg/kg). Serum tumor necrosis factor-alpha and nitrites were also determined to assess the mechanism of action of CNI-1493. Macrophage pacification decreased pancreatitis severity as determined by enzyme release and pancreatic histology score. Ascites volume and bronchoalveolar protein levels were also decreased, indicating that CNI-1493 prevents the loss of circulating blood volume and maintains hematocrit and mean arterial pressure, thus improving survival. CNI-1493 prevented the increase of serum tumor necrosis factor-alpha but not serum nitrites, implicating macrophage-derived cytokines and not nitric oxide in the pathogenesis of physiologic decompensation and death in this model of pancreatitis. PMID- 9744643 TI - Pharmacokinetics of a recombinant amino terminal fragment of bactericidal/permeability increasing protein (rBPI21) after liver surgery in rats and humans. AB - Major liver resections are associated with considerable morbidity and mortality. Gut-derived bacteria and bacterial endotoxin (LPS) are considered to play a central role in the pathophysiology of these complications. Like human BPI, rBPI21 binds to LPS from Gram-negative bacteria. By binding and clearing of LPS, rBPI21 can inhibit a number of endotoxin-induced humoral and cellular responses. Because of this capacity, rBPI21 could partially compensate for the loss of hepatic mononuclear phagocytic system function after liver resection. However, the liver is also thought to be an important organ for the clearance of BPI, and reduction of liver mass could result in a decreased clearance and exceedingly high plasma levels of rBPI21. In this study we therefore investigated the pharmacokinetics of rBPI21 in rats and in patients undergoing a major liver resection. Rats were administered an intravenous (i.v.) bolus of rBPI21 after undergoing a 60% or 80% hepatectomy (with sham-operated controls). Patients undergoing a hemihepatectomy and healthy volunteers received rBPI21 or placebo by continuous i.v. infusion for 48 h. Plasma concentrations were measured by sandwich ELISA. In rats, 60% hepatectomy did not consistently change the clearance of rBPI21, whereas 80% hepatectomy decreased the clearance of rBPI21 severalfold. In hemihepatectomized patients, the clearance of rBPI21 after major hepatectomy was also slower, when compared with healthy volunteers, but this difference had disappeared within 24 h. Our data indicate that the administration of rBPI21 in patients undergoing liver resection is well tolerated and does not result in exceedingly high plasma levels. Additional studies on the efficacy of rBPI21 in the prevention of complications after hepatectomy are needed. PMID- 9744645 TI - Central and regional hemodynamics during uncontrolled bleeding using hypertonic saline dextran for resuscitation. AB - The effects of hypertonic (7.5%) saline/6% dextran 70 (HSD) on central and regional hemodynamics were studied during uncontrolled intra-abdominal bleeding in 16 anesthetized pigs. Ultrasonic flow probes were placed proximally and distally to an aortic injury to indicate the incidence and extent of rebleeding after injecting 4 mL kg(-1) (N = 8) and 2.65 mL kg(-1) (N = 8) of HSD 10 min after the vascular injury was induced. The initial aortic bleeding reduced the blood flow rates to 71% of baseline in the skin, 53% in the splanchnic region, 42% in the upper aorta, and 15% in the kidney. Cardiac output dropped to 46% and the mean arterial pressure to 57% of baseline. The injection of HSD was followed by a prompt increase in all blood flow rates, but rebleeding started within 2 min in 13 of the pigs (81%). A second period of rebleeding occurred in six of them. The rebleeding averaged 300 mL, which is 62% of the blood lost when the aortic injury was induced. There was no significant difference between the treatment groups with respect to these blood losses or to the oxygen consumption, which was not restored by HSD. Five animals in each treatment group died after about 70 min, while the remaining six pigs (38%) survived the 120 min study period. These results suggest that HSD in the recommended dose, and even two-thirds thereof, promotes rebleeding when given shortly after a low energy intra-abdominal aortic injury. The fluid seems to have no beneficial effect on this type of uncontrolled hemorrhage. PMID- 9744646 TI - Tumor necrosis factor up-regulates intercellular adhesion molecule 1, which is important in the neutrophil-dependent lung and liver injury associated with hepatic ischemia and reperfusion in the rat. AB - Tumor necrosis factor (TNF) is released during hepatic ischemia/reperfusion (I/R) and plays an important role in the ensuing neutrophil-mediated lung and liver injury. Since TNF is not a direct neutrophil chemotaxin, we hypothesized that TNF may up-regulate neutrophil adhesion molecules, specifically intercellular adhesion molecule-1 (ICAM-1), following hepatic I/R, and that this molecule then plays an important role in tissue neutrophil influx. Rats underwent 90 min of lobar hepatic ischemia with reperfusion. Pulmonary and hepatic ICAM-1 expression were assessed by reverse transcription-polymerase chain reaction, Western blot analysis, and immunohistochemical staining. Increases in hepatic ICAM-1 were demonstrated within 1 h of reperfusion, while increases in pulmonary ICAM-1 were not seen until 6 h of reperfusion. Next, rats were treated with anti-TNF antibody or control antibody without TNF neutralizing properties prior to hepatic I/R. Pretreatment with anti-TNF antibody significantly decreased pulmonary and hepatic ICAM-1 expression after hepatic I/R. We next investigated the effects of pretreatment with anti-ICAM-1 antibodies on the lung and liver injury that follows hepatic I/R. Lung injury was assessed by changes in pulmonary capillary permeability as estimated by extravasation of Evans Blue dye and pulmonary neutrophil influx as measured by lung myeloperoxidase levels. Liver injury was assessed by hepatic neutrophil morphometrics and plasma liver enzymes (alanine aminotransferase). Pretreatment with anti-ICAM-1 antibodies significantly decreased pulmonary capillary permeability, pulmonary myeloperoxidase, hepatic neutrophil influx, and plasma alanine aminotransferase, as compared to animals pretreated with control antibody. These data suggest that TNF is a proximal trigger for pulmonary and hepatic ICAM-1 up-regulation following hepatic ischemia with reperfusion, and that ICAM-1 is important for pulmonary and hepatic neutrophil influx, with the resultant tissue injury, following hepatic I/R. PMID- 9744648 TI - Low molecular weight heparin alters porcine neutrophil responses to platelet activating factor. AB - Because platelet-activating factor (PAF) is an important mediator of inflammation and heparin has anti-inflammatory effects, we hypothesized that low molecular weight heparin (LMWH) would inhibit PAF-induced activation and chemotaxis in porcine neutrophils. Citrated blood was obtained from pentobarbital-anesthetized pigs, and neutrophils were isolated over a 55%/65% Percoll gradient. The effect of LMWH on basal phorbol myristate acetate (PMA)-induced superoxide (SO) release, as well as its effect on PAF priming for PMA-induced SO release, were investigated. Additionally, the effect of LMWH on PAF-induced chemotaxis of neutrophils across transwell membranes was evaluated. Baseline SO release in response to PMA was .351+/-.046 nmol/10(6) cells/min, and this was decreased to .289+/-.034 nmol/10(6) cells/min by pretreatment with 50 U/mL LMWH. PMA-induced SO production was increased by .240+/-.042 nmol/10(6) cells/min when cells were primed with 10 microM PAF. This priming effect of PAF was reduced significantly by pretreatment of neutrophils with LMWH at 10 and 50 U/mL. Chemotaxis of neutrophils in response to 100 microM PAF was significantly decreased to 70.02+/ 6.4% (n = 8) of the control response by pretreatment of cells with 50 U/mL LMWH. We conclude that LMWH has anti-inflammatory effects on porcine neutrophils, which includes attenuation of cell activation and chemotaxis in response to the lipid derived inflammatory mediator, PAF. PMID- 9744647 TI - Low molecular weight heparin is associated with greater cytokine production in a stimulated whole blood model. AB - Anticoagulants can influence production of cytokines in whole blood, but the effects vary depending on the type of anticoagulant and the immunological stimulus. We examined this further by anticoagulating normal blood with either unfractionated (UF) heparin or low molecular weight heparin (Fragmin) and stimulating each sample with lipopolysaccharide, zymosan A, phytohemagglutinin, immune complexes, H2O2, or RPMI. After incubating 24 h, the combination of lipopolysaccharide and Fragmin induced significantly greater concentrations of interleukin 1 (IL)-1beta (25+/-10 ng/mL; x +/- standard error), IL-8 (21+/-6), and tumor necrosis factor (TNF)alpha (.48+/-.24) compared with heparinized blood (p < .05). The combination of Fragmin and zymosan also induced significantly greater concentrations of IL-1beta (97+/-24) and TNFalpha (2.9+/-.8), but not IL 8 (2.0+/-15). Average levels of proinflammatory cytokines (IL-1beta, IL-6, IL-8, and TNFalpha) were greater with Fragmin anticoagulation for 36 of 40 comparisons, and patterns were similar for 6 h and 24 h incubations. Statistical difference was established in 33% of these comparisons. A composite score of proinflammatory cytokines for Fragmin-anticoagulated blood was significantly greater than expected for UF heparinized blood (p < .0001) after 24 h. Expression of one anti inflammatory cytokine (IL-10) was only slightly elevated for Fragmin anticoagulation. Proinflammatory cytokines are produced in greater quantities with Fragmin anticoagulation, which may be a disadvantage for ischemic conditions (e.g., cardiac surgery) but advantageous for long-term treatment of thrombosis. PMID- 9744649 TI - The influence of intestinal ischemia and reperfusion on bidirectional intestinal barrier permeability, cellular membrane integrity, proteinase inhibitors, and cell death in rats. AB - Intestinal ischemia and reperfusion injury (I/R) is probably involved in the pathogenesis of intestinal barrier dysfunction, associated with the concomitant translocation of enteric bacteria and toxins and the potential development of multiple organ failure. The intestinal endothelial and epithelial layers play a major role preventing the entry of toxic substances from the gut, but the influence of protease-antiprotease systemic balance on these barrier functions and the relationship between epithelial DNA synthesis, apoptosis, and endothelial and epithelial barrier macromolecule permeability are not fully investigated. Endothelial and epithelial barrier macromolecular permeability, epithelial DNA synthesis, the endothelial and epithelial plasma membrane system, apoptosis and oncosis, plasma levels of proteinase inhibitors, and proenzymes were measured in rats subjected to 20 and 40 min intestinal ischemia and 1, 3, 6, or 12 h reperfusion. Endothelial permeability increased after both 20 and 40 min intestinal ischemia. Epithelial permeability significantly increased during 1-6 h reperfusion after 20 min ischemia and during 1-12 h reperfusion after 40 min ischemia. Epithelial DNA synthesis increased in animals with 20 min ischemia followed by 12 h reperfusion. Plasma levels of prekallikrein, C1-esterase inhibitor, and alpha1-macroglobulin were significantly lower following both 20 and 40 min ischemia from 3 h reperfusion and on. Apoptotic epithelial cells significantly increased in animals subjected to 20 min ischemia followed by 12 h reperfusion. The severity of reperfusion injury in the intestinal endothelial and epithelial barrier seems to correlate with the period of ischemia and the pathway of cell damage and death, together with proteinase-antiproteinase imbalance. PMID- 9744650 TI - A novel fluid resuscitation therapy for hemorrhagic shock. AB - Fluid resuscitation is the usual therapy for hemorrhagic shock, and frequently consists of the infusion of large volumes of electrolyte solutions. However, to be successful, this therapy should be implemented soon after injury. A new treatment method in which the infusion could be delayed might result in a greater survival rate. Reducing the volume of fluid needed is also important. Both of these aspects of fluid resuscitation therapy were addressed in this study by supplementing the electrolyte solution with trans-sodium crocetinate (TSC). Rats were subjected to a severe hemorrhage, with 55% (or greater) of the estimated blood volume being removed over a period of approximately 10 min. There were five animals in each treatment group, and two types of experiments were done. In one, a bolus injection of TSC (or saline control) was given immediately after hemorrhage, followed 30 min later with an infusion of isotonic saline. In the other experiments, reduced infusion volumes of a TSC-saline infusion fluid were used. In both cases, TSC resulted in the survival of the animals while the controls all died. Whole-body oxygen consumption also increased with TSC, reaching 75% of the normal resting value after about 15 min. This correlates well with the increased survival rates seen, since mortality after hemorrhagic shock is associated with decreased oxygen consumption. These results suggest that the use of TSC could allow for later implementation of fluid resuscitation therapy as well as reducing the volume needed. PMID- 9744651 TI - Leukocyte-endothelial adherence correlates with pancreatic nitric oxide production in early cerulein-induced pancreatitis in rats. AB - The role of nitric oxide (NO) in microcirculation during the development of acute pancreatitis was not clear. An in vivo microscopic technique was used for evaluating leukocyte-endothelial adherence in the pancreatic microcirculation after induction (cerulein) of acute pancreatitis. Microdialysis was performed to detect pancreatic nitrate concentration (NO level) by high-performance liquid chromatography. Cerulein caused significantly reduced flow velocity in 1 h (31 %) and increased the number of sticking leukocytes in 2 h; both persisted for at least 3 h. Pancreatic NO level was found to be significantly elevated (2.5-fold) in 1 h and also persisted for 3 h. Both microcirculatory changes and NO elevation were significantly alleviated in cerulein-induced animals pretreated with NO synthase inhibitor (NG-nitro-L-arginine), indicating that elevation of NO could precede and account for a major portion of the observed microcirculatory changes. Furthermore, there was a strong positive correlation between numbers of adherent leukocytes and pancreatic NO level, suggesting that during the development of acute pancreatitis, NO could play an adverse role in microcirculation. PMID- 9744652 TI - Hemodynamic and metabolic responses to repeated hemorrhage and resuscitation with hypertonic saline dextran in conscious swine. AB - Previous work in our laboratory has demonstrated that HSD is an effective small volume resuscitation fluid for the treatment of hemorrhagic hypotension, but limitations to its usefulness in severe hemorrhage have not been explored. In the present study, animals (N = 12) were bled from an arterial line at a rate of 1 mL/kg/min until continuously monitored aortic blood flow was reduced to one-half its baseline value, and then they were immediately resuscitated with 7.5% NaCl/6% dextran 70 (hypertonic saline dextran, 4 mL/kg) administered intravenously over 3 min. After recording the maximum improvement in blood pressure, blood samples were obtained and the hemorrhage-resuscitation sequence was repeated until no further measurable increase in cardiac index or blood pressure could be elicited by resuscitation. In the majority of the animals, cardiac index and right and left ventricular stroke work could be improved at least through two bleedings and resuscitation. These improvements sufficed to increase oxygen delivery and consumption, despite the decreases in hematocrit induced by bleeding, transcapillary refill, and asanguinous fluid administration. Under these severe hemorrhage conditions, the acid-base imbalance was not improved by hypertonic saline dextran, and the rate of increase in acidosis was not affected by its administration. We observed a progressive decrease in base excess from +1.35+/ 3.19 (mean +/- standard error) to -12.9+/-2.1 mEq/L even when resuscitation improved oxygen consumption significantly by 95+/-20%. In animals that survived as many as three bleedings and resuscitation, the depletion of buffering capacity of the blood was most predominant, and bicarbonate reached a nadir of 7.62 mEq/L with a base excess of -22.4 mEq/L. It is evident that restoration of perfusion in shock treats only a portion of the physiologic dysfunction, leaving major metabolic derangements uncorrected. PMID- 9744653 TI - The future of medical technology assessment. PMID- 9744654 TI - Quality and its measurement. AB - The concept of assessing or measuring the quality of clinical care has to date not been an integral part of Australian hospital culture. Assessing and measuring quality of clinical care in a way that enables it to be quantified is an essential ingredient for quality improvement. While indirect measures, familiar enough in hospitals, are available they are rarely seen as techniques for assessing the quality of care. Technology that has now been available for the past 10-15 years involves making judgements by using medical record data compared with a matrix of outcome and process criteria. This approach makes quantification of quality possible. The methodology employed, Structured Quality Review (a term devised by Wilson and Goldschmidt), is described. Quality Standards in Medicine, a commercial product embracing Structured Quality Review, is described as a tool for assessing the quality of clinical care and measuring quality improvement. PMID- 9744655 TI - Health-care performance measurement systems and the ACHS Care Evaluation Program. Australian Council on Healthcare Standards. AB - The call for evidence-based medicine and information on health outcomes has brought with it performance measurement systems. The necessary attributes of these systems are now being addressed as are the attributes of the measures themselves. In this paper the Australian Council on Healthcare Standards Care Evaluation Program is reviewed in relation to these various attributes. The more focused systems should prove useful in achieving change in clinical practice. PMID- 9744656 TI - Complications of diabetes in the hospitalized population in Victoria, 1993-95. AB - A retrospective analysis of computerized data from the Victorian Inpatient Minimum Database (VIMD) was undertaken in order to describe the prevalence of diabetes and its associated complications in the hospitalized population over a 2 year period. While diabetes was rarely recorded as a principal cause of hospitalization (less than 0.5% of admissions), this condition was present in 4% (95,091) of the hospitalized population with a slight male excess. Cardiovascular disease was present in 60% of these diabetes-related admissions and was the principal diagnosis in a quarter of all cases. The prevalence of hypertension was 28%. Cardiovascular disease (CVD) was the principal diagnosis in 40% of in hospital deaths, and in women, the risk of CVD death was 22% greater than it was for men. Diabetes-related complications were noted in 22%; 3.3% recorded renal disease, 2.7% peripheral vascular disease, and ophthalmic and neurological complications were recorded in 2.1% and 1.4%, respectively. Of all lower limb amputations carried out in Victoria over the period, 40% (1281) were in people with diabetes. Eye surgery was carried out on (6.8%) 6463 diabetes-related separations. There are recognized limitations of using routinely collected computerized data. Nevertheless, data relating to number of amputations and eye surgery in those with diabetes can be used as indicators of the success of diabetes care and national strategies for prevention. PMID- 9744657 TI - Is asthma documentation improved by computer-facilitated data entry? AB - The documentation of acute asthma in written medical records was compared with data entered into a Computer-Assisted Triage System (CATS) in 104 children who presented to the emergency department and subsequently admitted to the Royal Alexandra Hospital for Children, Sydney. A total of 65 items in 5 categories were analysed and satisfactory documentation was defined as the recording of a specific item in more than 80% of records (written or electronic). Satisfactory documentation was observed for all 6 items in visit details and 9 out of 10 items in triage details for both recording systems. Nursing observations were better documented in the medical record than in CATS (87 vs 25%; kappa = 0.63). Documentation of medical details was also worse in CATS (75 vs 25%; kappa = 0.24) and the documentation of asthma severity was poor in both systems (31 vs 0%; kappa = 0.31). Attempts to improve asthma documentation through the development of a computerized medical record have highlighted further barriers to documentation. PMID- 9744658 TI - Organizing American grass roots physicians around quality: the Project Solo/Physicians Information Exchange experience. AB - Project Solo/Physicians Information Exchange has been the pioneer in multi-site community-based outcomes research. Quality is the foundation for this peer directed and developed grass roots organization that is a pioneer of Internet technology in its mission to provide physician leadership in health care. PMID- 9744659 TI - A classification for incidents and accidents in the health-care system. AB - Problems that arise from health-care management, rather than from a disease process, are now recognized as making a substantial contribution to patient morbidity and mortality and to the cost of health care. However, most classifications of these problems do not provide sufficient detail to allow comparisons or to develop better strategies for the prevention, detection and management of these problems. A 'Generic Occurrence Classification' was developed to record their salient features, place them in context and elicit any system or human error-based contributing factors. This was done by an iterative process in which 'natural categories', identified from over 2000 incidents and 800 adverse events, were placed in a hierarchical structure created using software written in Microsoft Visual Basic; data were stored in a Microsoft Access database. This was shown to be a valid and reliable way to compare incidents and accidents from different sources and to allow sufficient detail to be retrieved to develop preventive strategies. PMID- 9744660 TI - Central venous line disconnections with associated blood loss. AB - In 1996, a product trial was undertaken at Sydney Children's Hospital in response to an increasing number of central venous line disconnections associated with significant blood loss in small children. The product trialled was an extension line containing a one-way valve system with two side ports for needleless access- product code BC595, distributed by REM Systems. This product was chosen from a limited market for the compact size of the valve; this was important as it was to be secured onto the skin surface. It appeared that this product would prevent blood loss in the event of a line disconnection. PMID- 9744661 TI - Can we make it better? AB - The range and nature of patient complaints that resulted from the services provided in Southland base hospital over a 1-year period were studied. The data were assessed in the hope that they would provide new and important information to further develop quality assurance in the hospital service. There were 146 complaints, 15 of which were significant. Ten of these involved clinical care standards. There were 132,400 patient contacts during this time. The most common complaints related to the attitudes of health professionals, as perceived by the patient, and about information and other aspects that pertained to their individual care. These patient complaints did not provide unique information but would be useful if combined with other methods to determine patient dissatisfaction with the service provided. PMID- 9744662 TI - Evidence relating to goal planning in rehabilitation. PMID- 9744663 TI - Effect of transcutaneous electrical nerve stimulation (TENS) on Barthel Activities of Daily Living (ADL) index score following stroke. AB - OBJECTIVE: To evaluate the effectiveness of transcutaneous electrical nerve stimulation (TENS) and placebo TENS on the level of activities of daily living (ADL) of stroke patients. SETTING: A university hospital. PATIENTS: Patients who had had a stroke 30-240 days before entry to a university rehabilitation centre. DESIGN: Controlled design with block randomization and blinded assessment. INTERVENTION: All patients had Todd-Davies exercises. In group 1 (n = 30) TENS w th frequency of 100 Hz was used at an intensity that the patient could tolerate; n group 2 (n = 30) patients were given placebo TENS. The treatment protocol consisted of 40 sessions (eight weeks). OUTCOMES: The Barthel Index for daily living activities was used to measure functional changes over time, and the Ashworth Scale was used to measure spast city in the elbow, knee and ankle. These measurements were made prior to and following the treatment by assessors unaware of the patient's group allocation. RESULTS: There were 30 patients in each group. Patients in group 1 (active TENS) were more disabled at entry to the study. Statistically significant improvements were recorded in all parameters such as feeding, transfer, hyg ene, toileting, bathing, walking, climbing stairs, dressing, bowel and bladder care for group 1 (p<0.001) but only in some items in group 2. The change in total score was significant in both groups but the difference in the change score between the two groups was statistically significant (p<0.001). Spasticity was reduced in the active treatment group. CONCLUSION: TENS appears to be an effective adjunct in the regaining of motor functions and improving ADL in hemiplegic patients, but the accidental imbalance in severity of disability at entry makes interpretation uncertain. PMID- 9744664 TI - A preliminary report on the effectiveness of trunk targeting in achieving independent sitting balance in children with cerebral palsy. AB - OBJECTIVE: To assess the potential of Targeted Training in initiating or accelerating improved movement control of the trunk and hip joints in children with cerebral palsy so that independent sitting balance without specialized seating could be achieved. DESIGN: Six single case studies. SETTING: Assessment and review were undertaken in a specialized centre with intervention in the subjects' home or school. SUBJECTS: Children between the ages of two years five months and seven years five months (mean four years seven months) with an established diagnosis of cerebral palsy. None had independent sitting balance at the start of the study. INTERVENTION: Targeted Training using specialized equipment was directed at the appropriate few joints of the trunk as determined by initial testing and progressed when control at those joints had become automatic. The equipment provided support and challenged control learning. Periods of no intervention and placebo intervention, when the equipment was inappropriately set up, were also used. Two of the children ceased their traditional physiotherapy input while Targeted Training or placebo training took place. MAIN OUTCOME MEASURE: A new test was devised and validated to determine the most caudal extent of control of the vertical posture. In addition, a functional test of independent sitting balance was defined. RESULTS: All six children showed an increase in movement control and all gained independent sitting balance within 12-25 weeks (mean 16 weeks). This was irrespective of the continuation or cessation of traditional physiotherapy. CONCLUSION: These preliminary findings suggest that Targeted Training may be an effective means of promoting movement control and functional ability. Confirmation of these findings by other investigators would be of value. PMID- 9744665 TI - Outcome of strategy training in stroke patients with apraxia: a phase II study. AB - OBJECTIVE: Evaluation of a therapy programme for stroke patients with apraxia. The programme is based on teaching patients strategies to compensate for the presence of apraxia. This programme was designed for assessment and treatment by occupational therapists. DESIGN: The outcome was studied in a pre-post test design. Measurements were conducted at baseline and 12 weeks later. SUBJECTS: Thirty-three stroke patients with apraxia were treated at occupational therapy departments n general hospitals, rehabilitation centres and nursing homes. MAIN OUTCOME MEASURES: The following measurements were conducted: an apraxia test, a motor functioning test, observation of activities of daily living (ADL), Barthel Index, and an ADL questionnaire for the therapist and the patient. RESULTS: The patients showed large improvements in ADL functioning on all measures and small improvements on the apraxia test and the motor functioning test. The effect sizes for the disabilities, ranging from 0.92 to 1.06, were large compared to the effect sizes for apraxia (0.34) and motor functioning (0.19). The significant effect of treatment is also seen when individual improvement and subjective improvement are considered. Measured with the Barthel Index for instance, 71% of the patients improved. CONCLUSIONS: These results suggest that the programme seems to be successful in teaching patients compensatory strategies that enable them to function more independently, despite the lasting presence of apraxia. PMID- 9744667 TI - The use of one-arm crank ergometry in the prediction of upper body aerobic capacity. AB - OBJECTIVE: To determine whether a submaximal one-arm cranking test could be used to predict an individual's upper body aerobic capacity. This issue has potential importance for the fitness assessment of individuals with neurological disease or damage who have hemiplegia. METHODS: Nine healthy male volunteers (33+/-2.4 years) and nine female volunteers (27+/-1.9 years) performed a two-arm maximal, two-arm submaximal test and a one-arm submaximal arm crank ergometry test. Heart rate (HR) was monitored via a three-lead electrocardiogram (ECG) and expired air was analysed every 30 seconds throughout Prediction of peak oxygen consumption (Vo2peak) was calculated by linear extrapolation to an age-adjusted HRpeak. RESULTS: Heart rate and Vo2 were highly correlated in each test, and there were no significant differences between the Vo2peak values obtained from maximal crank ng and Vo2peak predicted from one- and two-arm submaximal tests for males and females. As expected, males were found to have significantly (p<0.001) higher actual and predicted Vo2peak values, indicating that separate regression equations should be used for males and females. CONCLUSIONS: Heart rate values obtained during one-arm submaximal cranking have the potential to predict arm cranking Vo2peak, and therefore provide an estimation of an individual's aerobic capacity, in addition to those obtained from the more traditional two-arm tests. PMID- 9744666 TI - The Northwick Park Dependency Score (NPDS): a measure of nursing dependency in rehabilitation. AB - BACKGROUND: Disability scores, such as the Functional Independence Measure (FIM) and Barthel Index, have been shown to correlate with care needs but cannot be used to assess them directly, as they do not indicate the number of people required to help with a task, nor the time taken. The Northwick Park Dependency Score (NPDS) is an ordinal scale that can be used to assess impact on nursing time. It takes 3-5 minutes to complete. Together with a short set of additional questions, it may be used directly to assess care needs in the community and to facilitate discharge planning. AIMS: To develop and evaluate the NPDS for use in a rehabilitation setting. METHODS: (1) DEVELOPMENT: Following a survey of existing instruments, tasks were selected on the basis of their impact on nursing time and divided into Basic Care Needs (BCN) and Special Nursing Needs (SNN). Cut off points were devised to reflect the number of helpers needed and time taken. Following evaluation of the NPDS version 5, minor changes were made to produce version 6 which was re-evaluated on a smaller scale. (2) EVALUATION: Inter-rater and intra-rater reliability were tested in a cohort of 23 inpatients using five senior nurses. Analysis included assessment of degree of association, significant differences, absolute agreement, and agreement +1 level. Although there is no gold standard, the BCN section should correlate inversely with independently assessed Barthel scores. Re-evaluation of version 6 was undertaken using the same method of analysis in a cohort of 21 patients using three senior nurses. RESULTS: On initial evaluation inter-rater reliability testing showed an excellent level of association in total composite score between each pair of nurses (rho = 0.73 0.92, p <0.01) and agreement +1 level for individual items ranged from 73 to 100%. Significant disagreements were in six items. On re-evaluation following minor modification, high levels of association were still seen for total BCN, SNN and composite scores both between and within raters, with very satisfactory levels of agreement for individual items. The BCN section of the NPDS showed good inverse correlation with Barthel scores (rho = 0.91, p <0.01). CONCLUSION: The NPDS is simple and practical to use in a busy setting. It is shown to be reliable and valid in its assessment of nursing dependency on the ward. Its translation into a directly costable measure of continuing care needs in the community now requires evaluation. PMID- 9744668 TI - The views of therapists on the use of a patient-held record in the care of stroke patients. AB - OBJECTIVE: To explore the views of therapists working with stroke patients on the use of a patient-held record (PHR) for stroke patients. A PHR was developed in the form of a pocket-sized booklet (21 cm x 14.5 cm) in which staff recorded information relating to the patient's management. The aim of the PHR was to facilitate communication and involve patients more directly in their care. METHODS: Six semi-structured group interviews were conducted with therapists (25 in total) from one inner city hospital. RESULTS: The following themes emerged from the content analysis: (1) Therapists were supportive of plans for a PHR, citing the benefits of greater patient involvement. (2) However, they questioned its feasibility, in particular the issue of patient responsibility and its use with the cognitively impaired. (3) They also questioned its ability to facilitate communication among health professionals because of existing differences in perspectives. (4) These therapists revealed concerns about the effect that information may have on patients. (5) They also raised practical issues about finding the time to make entries, wording and content of entries in the PHR. CONCLUSION: Responsibility for the PHR may enhance patients' understanding and involvement in their care, yet ownership alone does not guarantee the confidence needed to encourage dialogue between patients and care providers. Furthermore, it is doubtful whether a PHR can hope to overcome the fundamental differences in the philosophies of care which the therapists reported. PMID- 9744669 TI - A patient-centred study of the consequences of stroke. AB - OBJECTIVE: To explore subjective accounts of the consequences of stroke. DESIGN: Qualitative methods using depth interviews. PARTICIPANTS AND SETTING: Forty people sampled ten months post stroke from a hospital stroke register which was established in two adjacent health districts in North Thames Regional Health Authority. RESULTS: Interviewees reported a number of ways in which the stroke had affected their daily lives, including difficulty with leaving the house, doing the housework, pursuing former leisure activities, inability to walk in the way they wanted, problems with communicating, washing, bathing and dressing, and with confusion and deteriorating memory. In all these areas people described the loss of social contact that accompanied these changes, and the loss of valued roles which had been embedded in the everyday functions they had previously performed. In general, people over the age of 70 were more seriously affected. CONCLUSION: The type of changes which people reported would not easily have been captured using standardized outcome measures, pointing to the value of qualitative methods in providing subjective accounts. In terms of clinical practice, there is a need to reduce people's isolation after stroke by providing home visits after discharge, particularly to those living alone, and also by reducing disability through rehabilitation and by tackling the environmental obstacles which can imprison people in their homes. The findings suggest that many people with stroke would benefit from being able to talk about the changes which have occurred. Imaginative proposals are needed to develop ways to help replace the loss of activities, social contacts and social roles, particularly among older people with stroke. PMID- 9744670 TI - Association between amputation, arthritis and osteopenia in British male war veterans with major lower limb amputations. AB - OBJECTIVES: To investigate the association between amputation, osteoarthritis and osteopenia in male war veterans with major lower limb amputations. Specific questions were to determine whether lower limb amputees following trauma are at subsequent risk of developing osteoarthritis (OA) and osteoporosis of the hip on both the amputated and nonamputated sides. DESIGN: Retrospective cohort study in British Male Second World War veterans with major unilateral lower limb amputations. SUBJECTS: Seventy-five male Second World War veterans with major lower limb amputations known to be alive were invited to participate from a subregional rehabilitation centre. After exclusions, 44 agreed to attend for examination and radiological screening. METHODS: The presence of hip OA was determined from a single anterior posterior pelvic X-ray using two approaches: minimum joint space and the Kellgren and Lawrence (K&L) scoring system. Bone mineral density (BMD) was measured by a dual energy X-ray absorptiometry (DXA) scan and prosthetic rehabilitation outcome measures were recorded. RESULTS: Twenty-seven (61%) hips on the amputated side and 10 (23%) on the nonamputated side were positive for OA (based on Kellgren and Lawrence grade of >2). Using a minimum joint space threshold of below 2.5 mm, 24 (55%) hips on the amputation side and 8 (18%) on the nonamputated side were also positive for OA. There was a threefold increased risk of OA for those with above-knee compared to a below-knee amputation. By contrast, from published general population surveys only 4 (11%) cases of hip OA would have been expected on both the amputated and nonamputated hips. There was a significant decrease in femoral neck BMD in the amputated side (p <0.0001) and significantly lower BMD in above-knee amputees than in below-knee amputees (p = 0.0027) as compared to normal age- and sex-matched population. CONCLUSION: Male war veterans with unilateral major lower limb amputations develop significantly more osteoarthritis of the hip than expected on both ipsi- and contralateral sides. Amputation was also associated with loss of bone density. Above-knee amputees develop significantly more hip osteoarthritis and osteopenia of greater severity in the amputated side than below-knee amputees. PMID- 9744671 TI - Designing a Back Management Service: the Exeter experience and first year results. AB - BACKGROUND: The recent British Clinical Standards Advisory Group report on back pain highlighted the importance of primary care in the active management of acute back pain and, at the same time, recommended the establishment of rehabilitation services for those with subacute and chronic back pain. OBJECTIVE: To describe a new Back Management Service (BMS) in Exeter and to review the first year's results. METHODS: Analysis of observational data collected from the clinic and therapy database. ASSESSMENTS: Clinic outcome assessed by diagnostic and referral patterns as well as by general practitioner questionnaire. The Back Management Programme was assessed by Short Form 36 (SF36), Hospital Anxiety and Depression score (HAD) and Oswestry Disability Index (ODI). RESULTS: In the first year 297 patients were seen in the clinic; clinic triage resulted in confirmation of primary care diagnosis in 235 (79%) of patients, with diagnosis revised in 62 (21 %). One hundred and eighty-nine (63.6%) patients were referred for therapy. Of these, 54 were referred for one-to-one therapy and 135 were referred for multidisciplinary review with 29 patients subsequently recruited into the Back Management Programme for rehabilitation. Two out of three outcome measures were significantly improved at the end of the 36-hour, four-week course. CONCLUSIONS: A Back Management Service can provide useful diagnostic reassessment of patients with chronic low back pain, and can focus therapeutic effort for the effective management of their pain. PMID- 9744672 TI - High prevalence of hepatitis C virus genotype 6 among certain risk groups in Hong Kong. AB - The genotype of hepatitis C virus (HCV) of 172 HCV-RNA positive serum specimens taken from patients with chronic liver diseases, thalassaemia major, chronic renal failure (CRF), haemophilia and intravenous drug abusers (IVDA) was determined by analysis of the amplified 5'UTR region by genotype-specific oligonucleotide probes and restriction fragment length polymorphism (RFLP). Six different genotypes and subtypes (1a, lb, 2, 3, 4 and 6) were found. Genotype lb was the predominant genotype among patients with chronic liver diseases (69.6%), followed by genotype 6 (18.8%), which was similar to that reported for blood donors in earlier studies. Pronounced differences in the distribution of genotypes were seen between the four risk groups. Patients with CRF had a similar distribution to those with chronic liver diseases, whilst the greatest diversity of genotypes was seen in patients with haemophilia, which was expected since they were given factor VIII manufactured overseas. Genotype 6 was particularly prominent in patients with thalassaemia major (50%) and IVDA (62.5%). It is possible that clonal spread of HCV genotype 6 has taken place among a closed subset of the population in Hong Kong through intravenous drug abuse. PMID- 9744673 TI - The impact of the 1993 European revision of the AIDS case definition on back calculation estimates: an application in Italy. AB - OBJECTIVES: The revision of the case definition for AIDS within a given observation period causes discontinuity points on the epidemic curve to which back-calculation procedures can be sensitive. The aim of this work is to characterize the impact of the 1993 European revision of AIDS case definition and to evaluate the degree of distortion by which back-calculation estimates obtained on Italian data are affected. METHODS: The back-calculation procedure used, is a generalization of standard methods for estimating and projecting the AIDS epidemic. Age at onset of HIV infection, incubation time distribution, susceptible population and competitive non-AIDS mortality rates for infected persons are included in the estimation process. RESULTS: Back-calculation estimates resulted to be sensitive to the discontinuity point that the 1993 revision of AIDS case definition induced on the epidemic curve. The resulting effect is a tendency to overestimate HIV incidence in recent years by 20-40%. The maximum effect was found using data with end of AIDS cases at December 1993. The overestimation is a transient effect, that is, it is expected to almost disappear in 1995-1996. CONCLUSIONS: The introduction of the 1993 European revision of AIDS case definition caused distortion of back-calculation estimates in Italy, leading to overestimate incidence and prevalence of HIV infection for recent years. It is believed that this effect has occurred in all countries that adopted the 1993 European revision of AIDS case definition, and its characterization is potentially useful for further changes of AIDS case definition in the future. PMID- 9744674 TI - Temporal trends in reasons for and result of HIV-testing among women in Rome, Italy. AB - To describe the trend in the reasons for and result of women's HIV testing, systematic data was gathered for 11,523 consecutive women during pre-and post test visits at a major counseling and testing (CT) site of Rome, Italy, June 1985 July 1996. The number of tested women and the proportion of female clients attending the CT site significantly increased during the study period (p < 0.001), mostly because of reported sexual risk or when triggered by pregnancy. A significant increasing trend in the proportion of women who had one prior test (30% overall) was observed in all groups, apart from IDU. Newly diagnosed HIV infections were 319 (2.8%). The HIV prevalence was 27% in 1985-1987, when 66.7% of cases were IDUs, and decreased to 1.3% in 1994-1996, when 53.7% of cases were women reporting HIV infected partners. The findings suggest that information on the potential risk of HIV transmission has permeated the female population. The shift of newly diagnosed infections from IDUs towards women reporting sexual exposure, suggests the need for targeting preventive efforts to these population groups. Underlying reasons for multiple testing need further analysis. PMID- 9744675 TI - Presence of diabetes related complication at the time of NIDDM diagnosis: an important prognostic factor. AB - We studied the short-term natural history of patients with newly diagnosed non insulin dependent diabetes mellitus (NIDDM), and the prognostic role of history of NIDDM related complication at the time of first NIDDM diagnosis in relation to the development of a new complication or death. We performed a cohort study using data from the General Practice Research Database in the UK. We identified patients aged 30 to 74 years with a newly diagnosed NIDDM between 1990 and 1992 and followed them from the day of NIDDM diagnosis until June 1995. Among the 1077 patients identified, 437 (41%) developed a NIDDM complication during the follow up. NIDDM complications were more frequent among males and in the elderly. Sixty seven percent of the study cohort was initially free of any complication while the remaining 360 patients presented already one or more NIDDM complication at the time of their NIDDM diagnosis. History of diabetic related complication was associated with an increased risk of developing a new NIDDM complication (RR: 1.8; 95% CI: 1.5-2.2). Mortality was also greater among patients with history of NIDDM complication (RR: 1.5; 95% CI: 1.0-2.2). Patients with a history of any disorder related to diabetes before their clinical diagnosis of NIDDM are at increased risk of developing a NIDDM complication after the NIDDM diagnosis, as well as at increased risk of dying compared to diabetic patients with no history. PMID- 9744676 TI - Exploring the combined action of lifetime alcohol intake and chronic hepatotropic virus infections on the risk of symptomatic liver cirrhosis. Collaborative Groups for the Study of Liver Diseases in Italy. AB - Although alcohol intake and hepatitis B and C virus (HBV and HCV) infections are the major determinants of liver cirrhosis (LC) in western countries, the joint effect of these factors on LC risk has not yet been adequately studied. Data from three case-control studies performed in Italy were used. Cases were 462 cirrhotic patients admitted to Hospitals for liver decompensation. Controls were 651 inpatients admitted for acute diseases unrelated to alcohol. Alcohol consumption was expressed as lifetime daily alcohol intake (LDAI). Three approaches were used to explore the interaction structure. The Breslow and Storer parametric family of relative risk functions showed that an intermediate structure of interaction from additive to multiplicative was the most adequate one. The Rothman synergism index showed that the interaction structure between LDAI and viral status differed significantly from the additive model in particular for high levels of alcohol intake. When multiple regression additive and multiplicative models were compared after adjustment for the known confounding variables. a trend of the interaction structure towards the multiplicative model was observed at increasing levels of consumption. Better methods are needed for assessing mixed interaction structures in conditions characterized by multifactorial etiologies like cirrhosis of the liver. PMID- 9744677 TI - Strong regional links between socio-economic background factors and disability and mortality in Oslo, Norway. AB - STUDY OBJECTIVE: To study geographical differences in mortality and disability and socio-economic status in Oslo, Norway. SETTING: A total of 25 local authority districts within the city of Oslo. DESIGN: Analysis of age adjusted mortality rates aged 0-74 in the period 1991-1994, and cross sectional data on disability pensioners aged 50-66 and socio-economic indicators (low education, single parenthood, unemployment, high income) in 1994. MAIN OUTCOME MEASURES: The levels of correlation between the health outcomes (mortality and disability) and socio economic exposure variables. MAIN RESULTS: The geographical patterns of mortality and disability display substantial similarities and show strong linear correlation with area measures of socio-economic deprivation. The ratios between the highest and lowest area mortality rates were 3.3 for men and 2.1 for women, while the high-low ratios of disability were 7.0 for men and 3.8 for women. For women deprivation measures are better correlated with disability than mortality. While disability and mortality display similar correlations with deprivation measures for men. CONCLUSIONS: The social gradients in health are substantial in Oslo. Further ecological analysis of cause specific morbidity and mortality and the distribution of risk factors ought to be done to identify problem areas suitable for interventions. However, to understand the mechanisms and the relative importance of each etiological factor, studies based on individual data have to be performed. PMID- 9744678 TI - A space-time criterion for early detection of epidemics of influenza-like illness. AB - OBJECTIVES: To develop a method based on a space-time criterion for early detection of epidemics of influenza-like-illness in France. METHODS: Since 1984, the French Communicable Diseases computer Network (FCDN) routinely detects epidemics of influenza-like-illness when the national incidence rate is, for 2 consecutive weeks, above a threshold computed by a periodic regression model. It appears that some areas reported early increases in incidence several weeks before the national epidemic. An optimised space-time criterion allows an early detection of the epidemic periods. RESULTS: Applying this space-time criterion to the last 11 epidemics (from 1986), the sensitivity was 0.82 and the specificity was 0.99. CONCLUSION: This simple procedure can be used as an additional tool for early detection of an epidemic taking into account the distribution of new cases in space and time. PMID- 9744679 TI - Reported influenza in pregnancy and childhood tumour. AB - The present study was conducted to test the hypothesis that exposure to influenza in pregnancy increases the risk of tumour of certain type in childhood. Children ages 17 years or less diagnosed in Greece with brain tumours or neuroblastomas from 1982 to 1993 (n = 94) were contrasted to 210 controls selected from the same hospitals. Mothers of these children were interviewed about a variety of possible etiologic factors. The prevalence of influenza in Greece for each year during the period 1984-1992 was also compared with the number of children born during the same year who subsequently developed brain tumour or neuroblastoma. The results indicate a significant association between influenza in pregnant women and occurrence of tumour in index child (OR: 3.15, 95% CI: 1.13-8.77). These results persisted when adjustment for potential confounding factors was made. The findings should be interpreted cautiously because of lack of serologic documentation of information about infection obtained in interviews. A positive correlation (r = 0.74) of the number of tumour births by year of birth with the prevalence of influenza during the same year was also noted. This exploratory study is one of the few case-control studies of the epidemiology of childhood tumours in children, and the results suggest directions for future epidemiologic studies in this relatively uncharted field. PMID- 9744680 TI - Hyperlipidaemia: differences in management practices and attitudes in two regions in Europe--Sicily and the Stockholm area. AB - In order to compare attitudes and management concerning hyperlipidaemia and risk factors for coronary heart disease among doctors in northern and in southern Europe, a questionnaire study was undertaken among doctors in primary health care and departments of internal medicine in Sicily and Stockholm. The regions differed in culture and health-care structure. Guidelines were similar, but screening of healthy individuals was recommended in Sicily, and not in Sweden. One hundred and fifty-three general practitioners in Sicily and 120 in Stockholm, 211 internists in Sicily and 83 in Stockholm participated. Main outcome measures were management policies for investigation and treatment and also attitudes. Routine lipid checks at first visits were done by few doctors in Stockholm but by a majority in Sicily (p < 0.001); in the presence of general cardiovascular risk factors (other than heredity, diabetes, cardiovascular disease and hypertension), routine checks were carried out more often by both general practitioners (p < 0.001) and internists (p < 0.005) in Stockholm. Drug treatment was initiated at lower cholesterol levels for secondary and primary intervention, cardiovascular disease, cardiovascular risk factors and hereditary hyperlipidaemia by both groups in Sicily (p < 0.001), as was dietary treatment. Secondary prevention was considered important by all groups, but primary prevention only by Sicilian doctors. We concluded that there were differences in views and management practice between doctors in Sicily and in Stockholm on the investigation and treatment of patients with hyperlipidaemia. Doctors tested lipids at first visits in Sicily but not in Stockholm. Treatment was initiated at lower levels of cholesterol in Sicily. PMID- 9744682 TI - Prevalence of Helicobacter pylori infection in two Spanish regions with different incidence of gastric cancer. AB - It is a cross-sectional study, comparing the prevalence of Helicobacter pylori infection (prevalence of IgG antibodies to H. pylori) in the healthy population of Ubrique and Grazalema (mountain location, mortality from stomach cancer 20/100,000) and in Barbate, (coastal location, mortality from stomach cancer 10/100,000) in the province of Cadiz, southern Spain. The subjects were randomly selected, 163 men and 169 women, 18 years or older; 179 persons were studied in the inland, and 154 in the littoral in January 1997. Of the 332 subjects investigated, 43% were positive, a mean antibody titer of 337 IU/1 (95 % CI: 254 420), and 56% were negative, with a mean titer of 18 IU/1 (95% CI: 15-19). In the coastal population, 30% has positive titers and 54% in the mountain location. By age: 18-40 years, 30% of littoral and 41% of inland population had positive titers; 41-60 years, 35% of those living in the littoral and 58% of inland population had positive titers; > 60 years, 24% of coastal inhabitants and 62% of those living in the inland had positive titers. Living in mountain locations in the province of Cadiz involves a greater ecological risk for H. pylori infection (p < 0.05). PMID- 9744681 TI - The epidemiology of bite and scratch injuries by vertebrate animals in Switzerland. AB - Pet and wildlife populations are a potential source of various public health problems, and injuries and complications due to animal bites and scratches are the most obvious. As no population based data on the frequency of animal bites were available at a national level in Switzerland, a study was conducted by the Swiss Sentinel Surveillance Network. The objectives of this study were to estimate the incidence of medical consultations due to bite and scratch injuries in humans caused by vertebrate animals, to identify possible risk factors, and to assess bite management habits in primary health care. An annual bite and scratch incidence rate of 325 per 100,000 population was estimated. Consultations peaked during the summer months and geographical differences in the reported incidence were observed. Dogs accounted for more than 60% and cats for about 25% of all cases reported. Animal bites and scratches were frequent in persons under 20 years of age. In most ages, the incidence was higher among women than among men, but not in children under the age of ten years. The incidence of cat bites was especially high in adult women. Bites to the head and neck were most frequent in infants and young children and accounted for approximately one third of the reported cases in this age group. Patients sought medical care principally for primary wound care (52.0%) and for vaccination advice (29.6%). Rabies postexposure prophylaxis was initiated in 1.1% of patients. Wound infection was reported in 10.9% of cases, with cat bites/scratches being more often infected than injuries due to dogs. Hospitalization was reported in 0.3 % of patients. Data from the emergency department of two district hospitals showed that head and neck injuries were more frequent in out-patients and a higher proportion of persons presented with wound infections (14.1%). The hospitalization rate for emergency department visits was 4.7%. Animal bites and scratches are common events in Switzerland. They represent a public health issue of growing importance due to the steadily increasing pet population. A practice based sentinel surveillance system may be an appropriate tool to monitor national trends in animal bites and scratches. PMID- 9744683 TI - An epidemiological study of pityriasis rosea in the Eastern Anatolia. AB - The purpose of this study was to investigate the epidemiological features of pityriasis rosea (PR) in the Eastern Anatolia, Turkey. Three hundred ninety-one patients (214 females, 177 males) with PR seen during the years 1992-1995 were analyzed for annual incidence among dermatologic outpatients, sex, age, and distribution by month and year. The average annual incidence was 0.75 per 100 dermatologic patients. PR was reported to be slightly more common in women by margin of 1.2:1.0. Eighty-seven percent of the cases were between the ages of 10 and 39 years, with a peak in the 20-29 age group. The incidence of the disease was much higher in the rainy and snowy months. No declining incidence was observed over the years. Changes in incidence from year to year, though not great, were statistically significant. PMID- 9744684 TI - A multicentre serosurvey on diphtheria immunity in a French population of 1004 subjects. AB - Diphtheria immunity was determined in serum specimens obtained in 1994 from 1004 subjects seen in emergency departments of three distant French cities. An enzyme immunoassay was used to measure serum diphtheria antitoxin concentrations according to the following criteria: (a) antitoxin < 0.01 IU/ml: susceptibility, (b) 0.01-0.09 IU/ml: basic protection, (c) > or = 0.10 IU/ml: full protection. Among these patients, 20.3% were fully susceptible to diphtheria, 30.3% had basic but doubtful protection and only 49.4% were fully protected. Protection was different by age-groups: 73.5% of the subjects under 40 years of age, 46% between 40 and 65 and 33% over 65 were fully protected. Protection decreased with increasing age (p < 0.001)and was greater for men than women after 40 years of age (p < 0.001). The results of this exploratory study indicate that the enhancement of diphtheria immunity by boosters in adult population should be reconsidered in France as well as in many industrialized countries. PMID- 9744685 TI - Prevalence of Chlamydia pneumoniae specific antibodies in different clinical situations and healthy subjects in Izmir, Turkey. AB - Serological markers for Chlamydia pneumoniae were investigated by using the microimmunofluorescence (MIF) test in various age and patient groups in a specific area in Turkey. IgG seropositivity to C. pneumoniae was 64.3% and 18.7% in healthy adults and children, respectively. The highest positivity rate (77%) was in the 15-19 age group. Among the groups investigated, serological findings revealed a possible etiological association between C. pneumoniae and the clinical condition in the groups with acute myocardial infarction, atypical pneumoniae and chronic obstructive pulmonary disease. PMID- 9744686 TI - Antigenic and genomic analysis of a Borrelia burgdorferi sensu stricto strain isolated from Ixodes ricinus ticks in Alto Adige-South Tyrol, Italy. AB - A Borrelia burgdorferi sensu lato strain isolated from IXodes ricinus ticks in Alto Adige-South Tyrol (Northern Italy) was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of whole cell proteins, Western immunoblotting analysis (WBA) with polyclonal and monoclonal antibodies, and pulsed-field gel electrophoresis (PFGE). The isolate named BZ6 was identified as belonging to the genospecies B. burgdorferi sensu stricto on the basis of its protein profile and its reactivity with monoclonal and polyclonal antibodies. The PFGE study performed with the two rare-cutting restriction enzymes MluI and SmaI confirmed the SDS-PAGE and WBA characterizations, but showed a genetic diversity between the isolate and two out of the three B. burgdorferi sensu stricto strains used in this study as controls, the American type strain B31 and the locally isolated strain BZ1. No difference in the PFGE patterns between the isolate BZ6 and the Swiss strain IRS was noted. Our findings show the value of PFGE analysis for classifying B. burgdorferi sensu lato isolates and for revealing their genetic diversity, and its usefulness for epidemiological investigations. PMID- 9744687 TI - Failure to recognize rapidly growing mycobacteria in a proficiency testing sample without specific request--a wider diagnostic problem? AB - Fifty participants in the Swiss External Quality Control Program in Bacteriology/Mycology received a diagnostic sample containing Mycobacterium fortuitum. Only 31 used some sort of acid-fast stains, and 13 reported the diagnosis of M. fortuitum or rapidly growing mycobacteria. We conclude that the presence of 'rapid growers' in routine bacteriology samples is underestimated, and that acid-fast stains should be performed on suspicious Gram-positive rods. PMID- 9744688 TI - Newly diagnosed HIV infections among pregnant women and their partners. PMID- 9744690 TI - Halothane-induced acute liver failure: continuing occurrence and use of liver transplantation. AB - BACKGROUND/AIMS: This study was aimed at determining if the frequency and pattern of acute liver failure (ALF) following halothane anaesthesia had decreased during the last 11 years in comparison with a previous series of 48 patients referred between 1965 and 1984 and whether clinical outcome had been altered by the introduction of liver transplantation. METHODS: Between January 1985 and December 1995, all patients with halothane-induced ALF admitted to the Liver Failure Unit at King's College Hospital were identified. Four other European liver transplant centres with a known interest in acute liver failure also provided data. RESULTS: Of the 18 patients admitted, the clinical data were complete in 15. Ten of these patients had at least one previous halothane anaesthesia with documented clinical complications following the earlier exposure in six. Four patients had been re exposed to halothane within 1 month of the penultimate halothane anaesthesia. Of the 15 patients four survived with medical management alone and 11 patients fulfilled transplant criteria. Four of the latter group were not listed because of rapidly deteriorating medical state and died, and of the seven patients who were listed, three died without a liver becoming available and four were transplanted, one of whom survived. No patient who had grade 4 encephalopathy and a prothrombin time > 50 s survived without a transplant. The survey of the other European liver centres recorded a total of 19 patients with halothane-induced ALF including three cases reported in the literature. Of those, 13 patients had been transplanted with nine survivors. CONCLUSION: Cases of halothane-induced acute liver failure still occur, albeit at a lower frequency than previously, and the Committee on Safety of Medicines guidelines are not being followed. The results of transplantation in these patients are encouraging. PMID- 9744689 TI - Halothane hepatitis. AB - Halothane, an effective and usually safe anaesthetic agent, is rarely associated with the development of fulminant hepatic failure. Guidelines have been developed to reduce the probability of a patient developing halothane hepatitis. However, cases continue to occur and, in some cases, the guidelines have been ignored. Stricter adherence to the guidelines will reduce, but not totally prevent, further cases from occurring. Once halothane hepatitis has developed, there are no specific treatments and liver replacement may be required. Halothane hepatitis is a paradigm for immune mediated adverse drug reactions. The mechanism appears to be related to development of sensitization to both autoantigens (including CYP2D6) and halothane-altered liver cell determinants. PMID- 9744691 TI - The detection and localization of inducible nitric oxide synthase production in the small intestine of patients with coeliac disease. AB - OBJECTIVES: To investigate whether there are increased numbers of inducible nitric oxide synthase (iNOS) containing cells in the small intestine of patients with coeliac disease and the localization of nitric oxide synthase production. DESIGN: Small intestinal biopsy specimens from patients with coeliac disease (11 untreated, 10 treated) and nine disease controls were studied. METHODS: Histochemical staining of sections for NADPH-diaphorase activity was performed, which gives an indication of NOS activity. iNOS protein was detected with immunohistochemistry and iNOS mRNA expression was detected using in situ hybridization with an oligonucleotide probe cocktail for iNOS. Cell phenotype was detected using monoclonal antibodies to CD3 (T-lymphocytes) and CD45 (total inflammatory cell infiltrate). RESULTS: There was significantly greater NADPH diaphorase staining in the lamina propria of patients with untreated coeliac disease (P < 0.005). The same pattern was found for immunohistochemical and in situ hybridization methods of staining for iNOS in each of the patient groups (P < 0.005) but no epithelial staining was seen with any method. The pattern of iNOS staining in the lamina propria appeared in a similar distribution to that of the inflammatory cell infiltrate. At least 80% of the significantly increased total inflammatory cell infiltrate (CD45) in the lamina propria of patients with untreated coeliac disease was lymphocytic (CD3) whilst the iNOS staining cells made up less than 15% of the total inflammatory cell infiltrate. CONCLUSIONS: There is a significant increase in the number of NOS staining cells of the inducible isoform in the lamina propria of patients with untreated coeliac disease. The lamina propria and not the epithelium is the site of iNOS production in coeliac disease. It appears that inflammatory cells other than T-lymphocytes are likely to be the cellular sources of iNOS production within the lamina propria. This is the first study to demonstrate increased numbers of iNOS producing cells in the small intestine of patients with untreated coeliac disease and suggests a role for nitric oxide in the pathogenesis of the histological changes seen in coeliac disease although it may be a non-specific inflammatory response to immune activation by gluten in susceptible individuals. PMID- 9744692 TI - Sexual behaviour in untreated and treated coeliac patients. AB - BACKGROUND: Sexual behaviour is often altered in chronic illness. AIM: To evaluate sexual behaviour in coeliac patients before and after treatment with a gluten-free diet. PATIENTS: Fifty-five adults with coeliac disease and 51 age- and sex-matched healthy controls. METHODS: Routine clinical and laboratory work up was used for diagnosis of coeliac disease. Age of first sexual intercourse, prevalence of individuals who were sexually active, frequency of intercourse, reduction in sexual desire, difficulty in attaining orgasm, pain during intercourse, and prevalence of individuals defining themselves as satisfied with their sexual life were investigated by an anonymous, self-administered questionnaire administered before and after one year's treatment with a gluten free diet in coeliac patients, and only once in controls. Analyses included clinical conditions, demographic and socio-economic data. RESULTS: Compared with controls, untreated coeliac patients had a significantly lower frequency of intercourse and a lower prevalence of individuals satisfied with their sexual life. Patients with overt and subclinical coeliac disease did not show significant differences for any indices of sexual behaviour. Compared with untreated conditions, coeliac patients after one year of treatment had improved values for all indices of sexual behaviour: differences were significant for frequency of intercourse and prevalence of individuals satisfied with their sexual life. CONCLUSION: Untreated coeliac disease, even in its subclinical presentation, is associated with disorders in sexual behaviour which are improved by the dietary treatment. PMID- 9744693 TI - Metal stents improve dysphagia, nutrition and survival in malignant oesophageal stenosis: a randomized controlled trial comparing modified Gianturco Z-stents with plastic Atkinson tubes. AB - OBJECTIVE: To compare modified Gianturco metal stents with plastic Atkinson tubes in the palliation of malignant dysphagia. DESIGN: Patient single-blind, multi centre prospective, randomized trial. SETTING: Three district general hospitals in the Wessex region. PARTICIPANTS: Thirty one consecutive patients with inoperable malignant oesophageal stenosis causing dysphagia and suitable for treatment with an endoprosthesis. INTERVENTIONS: Patients were randomized to receive either a modified Gianturco metal stent or a plastic Atkinson tube. Sedation was similar and patients were given identical dietary advice. Data were collected after insertion until the patients' death. MAIN OUTCOME MEASURES: Procedural mortality/morbidity; hospital stay; weight loss; quality of life (Nottingham Health Profile, Spitzer QL index and specific questions about dysphagia and enjoyment of food); duration of survival after insertion; cost effectiveness of each intervention. RESULTS: Overall complication rates were similar in the two groups. Compared with Atkinson tubes, patients with Gianturco stents had better palliation of dysphagia (median dysphagia score 1 vs 2, P = 0.04), maintained their weight longer (median percent weight loss 0.66 vs 6.51, P = 0.007), enjoyed food more (enjoyment score 2 vs 1, P = 0.03) and survived longer (log rank P < 0.025). Patients with metal stents were discharged from hospital earlier (Gianturco 4 days, Atkinson 10 days, P = 0.001), and initial treatment cost was lower if the cost of hospital stay exceeded pound sterling 120 per day. CONCLUSION: Gianturco stents are superior to Atkinson tubes in the palliation of malignant oesophageal stenosis. PMID- 9744694 TI - The role of mucosal mast cell degranulation and free-radical generation in intestinal ischaemia-reperfusion injury in rats. AB - OBJECTIVES: In this study, we determined the role of mucosal mast cell (MMC) activation in the pathogenesis of intestinal ischaemia-reperfusion (I/R) injury by immunohistochemical analysis using anti-RMCP II antibody. In addition, we investigated the role of free-radical generation in the activation of MMCs in this model. METHODS: In the first experiment, rats were divided into four groups: (1) sham operated; (2) I/R + saline; (3) I/R + the mast cell stabilizer, MAR-99 (30 mg/kg); and (4) I/R + MAR-99 (100 mg/kg). Treatment with MAR-99 was started 1 h before the occlusion of the superior mesenteric artery (SMA). In the second experiment, rats were divided into five groups: (1) sham operated; (2) I/R + saline; (3) I/R + superoxide dismutase (SOD; 50,000 U/ml); (4) I/R + catalase (90,000 U/ml); and (5) I/R + allopurinol (50 mg/kg/day). Intravenous administration of SOD and catalase was performed 1 h before SMA occlusion. Oral administration of allopurinol was started 2 days before I/R surgery. We measured several parameters of intestinal mucosal injury and evaluated the degranulation of MMCs by using an immunohistochemical technique. RESULTS: The number of resting MMCs, detected by anti-RMCP II antibody, was significantly decreased in the I/R treated rats. The I/R treatment induced a decrease in the mucosal histamine content and an increase in plasma histamine levels. Mucosal permeability in the small intestine was significantly enhanced by I/R treatment. However, these changes were significantly prevented by pretreatment with the MMC stabilizer, MAR 99. Furthermore, administration of several free-radicals scavengers (SOD, catalase, and allopurinol) also blocked the I/R-induced degranulation of MMCs. CONCLUSION: These data indicate that activation of MMCs was involved in the pathogenesis of I/R-induced intestinal mucosal injury. In addition, some parts of the I/R-induced MMC activation pathway were mediated by free-radical generation. PMID- 9744695 TI - Risk factors for rebleeding and fatal outcome in elderly patients with acute peptic ulcer bleeding. AB - OBJECTIVE: Mortality after peptic ulcer bleeding (PUB) is high in elderly patients despite therapeutic advances. Little is known about what actually determines rebleeding and mortality. The objective of this study was to investigate which factors may have an independent influence upon rebleeding and mortality in patients with PUB. DESIGN: Prospective cohort study. PARTICIPANTS: Patients, above 60 years of age, hospitalized due to an endoscopically verified acute PUB were included in the study (n = 508). INTERVENTIONS: The occurrence of rebleeding within 3 days and mortality within 30 days was registered for all patients. A predefined set of variables with a potential to influence rebleeding and mortality was analysed in a multiple logistic regression model. MAIN OUTCOME MEASURES: Odds ratios (with confidence intervals) for all predefined variables with respect to influence upon rebleeding and mortality, respectively. RESULTS: The risk of rebleeding was significantly increased with greater age and if the patient was suffering from shock, while omeprazole infusion, acetylsalicylic acid (ASA)/non-steroidal anti-inflammatory drug (NSAID) intake before admission and gastric ulcer localization were associated with a lower risk. Mortality was significantly increased with greater age, heart disease and blood pressure < 100 mmHg at admission. Previous ulcer history and the presence of a Forrest class IIa ulcer significantly reduced this risk. CONCLUSIONS: Elderly patients in shock admitted due to their first peptic ulcer bleeding run the greatest risk of an unfavourable outcome. PMID- 9744696 TI - Somatostatin in the prevention of recurrent bleeding after endoscopic haemostasis of peptic ulcer haemorrhage: a preliminary report. AB - OBJECTIVE: Although endoscopic injection therapy provides excellent initial haemostasis in actively bleeding ulcers, the incidence of recurrent haemorrhage is not negligible. The aim of this study was to compare somatostatin, omeprazole and ranitidine in preventing further haemorrhage after endoscopic injection haemostasis. METHODS: Seventy-three patients with major stigmata of ulcer haemorrhage at endoscopy were treated with epinephrine injection and randomly assigned to receive either omeprazole (n = 24) or ranitidine (n = 24) or somatostatin (n = 25). The three groups were similar in all background variables including mean age, clinical and endoscopic features, severity of bleeding and timing of the haemostatic procedure. All patients underwent a second endoscopic look at 48 h. Failures of treatment or retreatment underwent emergency surgery. RESULTS: There were no statistically significant differences between the groups in terms of initial haemostasis, need for emergency surgery, transfusion requirements, length of hospital stay or mortality. Early recurrent haemorrhage was 5/22 (22.7%) in the ranitidine group, 5/23 (21.7%) in the omeprazole group and 2/23 (8.7%) in the somatostatin group. No major side-effect was noted with drug therapy. CONCLUSIONS: The preliminary results suggest that somatostatin might be more effective than ranitidine and omeprazole in the prevention of recurrent haemorrhage following endoscopic injection therapy of bleeding peptic ulcers. PMID- 9744697 TI - Gastric meal accommodation and symptoms in diabetes. A placebo-controlled study of glyceryl trinitrate. AB - OBJECTIVE: To study mechanisms behind postprandial symptoms in patients with diabetes mellitus and the effect of nitric oxide (NO) on gastric accommodation and symptoms in these patients. DESIGN: A double-blind, placebo-controlled, randomized trial was designed in 20 patients with type 1 diabetes (10 male and 10 female, aged 35.3 +/- 7.6 years). METHODS: 0.5 mg sublingual glyceryl trinitrate (GTN), a donor of exogenous NO, or placebo was administered 5 min prior to a 500 ml soup meal. Gastric accommodation of the meal was assessed by abdominal ultrasound. Accommodation in proximal stomach was visualized in a sagittal area (Psa) and a frontal diameter (Pfd) and accommodation in distal stomach was visualized in a sagittal area of the antrum (Asa). Symptoms were assessed using visual analogue scales. RESULTS: Psa correlated significantly (r = 0.57, P = 0.015) with perception of fullness 5 min after the meal, whereas Pfd correlated significantly (r = 0.67, P = 0.004) with nausea at 15 and at 25 min after the meal. Asa correlated (r = 0.50, P = 0.05) with pain at 5 min, 10 min (r = 0.50, P = 0.05) and 25 min (r = 0.68, P = 0.007). GTN had no significant effect on Psa or Pfd, but reduced significantly (P = 0.05) Asa (1 3.5 +/- 4.5 cm2 with GTN vs 16.1 +/- 4.3 cm2 with placebo). GTN increased significantly (P = 0.04) the intragastric proximal/distal meal distribution ratio (proximal/distal sagittal area), but had no significant effect on symptom scores. CONCLUSION: In patients with diabetes, a large proximal stomach is associated with perception of fullness and a large antrum is associated with perception of pain after a meal. Sublingual administration of GTN prior to the meal decreases the antral area and improves the intragastric meal distribution, but fails to improve symptoms. PMID- 9744698 TI - Detection and typing of the virulence determinants cagA and vacA of Helicobacter pylori directly from biopsy DNA: are in vitro strains representative of in vivo strains? AB - BACKGROUND: The relationship of Helicobacter pylori genotypes to gastrointestinal disease has relied on cultured isolates. This assumes that cultured strains are representative of in vivo strains. OBJECTIVE: To detect and type the cagA status and the vacA genotypes directly from biopsy DNA without the need for culture, and to further define the relationship between H. pylori genotypes and gastroduodenal pathology. METHODS: Fifty-two Caucasian patients undergoing routine endoscopy for dyspepsia had additional biopsies taken from four gastric sites and one duodenal site for biopsy DNA preparation. An antral sample was taken for biopsy culture. All recruited patients were H. pylori-positive on rapid urease test for Campylobacter-like organisms (CLO test) and/or histology. By polymerase chain reaction (PCR), the cagA status and the vacA s and m types were detected directly from biopsy DNA. RESULTS: H. pylori isolates were cultured from 28/52 patients in whom infection was detected by PCR. Two isolate types differed from biopsy types. Fifty of the 52 patients, strains were typable from all four gastric sites and in 51/52 the same strain predominated throughout. The cancer strains were all cagA positive/vacA s1 type. There was a correlation between cagA positivity and vacA s1 (41/43). There was no difference between the cagA-positive/vacA s1 strains and the presence or absence of ulcers. There were only 5/52 vacA s2 m2 and four were in the non-ulcer dyspeptic group. CONCLUSION: cagA status and the vacA genotyping was successful with tissue samples taken directly from gastric and duodenal biopsies. Discrepancies between isolate and biopsy strain types stress the need for caution when interpreting in vitro strain types and suggest that direct PCR of biopsies is the preferred typing technique. The cagA status and the s1 vacA allele are unreliable as single markers in determining the risk of developing peptic ulcer disease. PMID- 9744699 TI - A strategy for osteoporosis in gastroenterology. AB - Osteoporotic fractures are a major public health problem. Gastroenterologists see many patients at risk of osteoporosis, particularly those with coeliac disease and inflammatory bowel disease. In this paper, the extent of the problem is reviewed and a strategy of investigation and treatment is recommended. PMID- 9744700 TI - Octreotide in the treatment of intestinal lymphangiectasia. AB - Primary intestinal lymphangiectasia is characterized by dilated small bowel lymphatics and loss of lymph into the bowel lumen resulting in hypoproteinaemia and oedema. Some patients have a more generalized lymphatic abnormality associated with lymphoedema of the limbs and chylous pleural effusions. There is no specific treatment although enteric protein loss may decrease with a low-fat diet. This report describes a patient with severe primary intestinal lymphangiectasia, associated with limb oedema and recurrent pleural effusions, who responded to treatment with octreotide. Before starting octreotide she required weekly intravenous albumin infusions to maintain the serum albumin above 20 g/l. Bilateral pleural effusions repeatedly reaccumulated despite pleurectomy and subsequently tetracycline pleurodesis. Treatment with octreotide, 200 microg twice daily, resulted in a reduction in enteric protein loss from 16 to 4.1% in 5 days (normal less than 1%) and the serum albumin was maintained between 22 and 26 g/l without the need for albumin infusion. Oedema in the arms resolved completely and the pleural effusions did not reaccumulate. The mechanism of action of octreotide in this condition appears to be due to a reduction in gut protein loss and another, as yet unidentified, action. PMID- 9744701 TI - Pancreatic carcinoma in remnant pancreas after pancreatectomy for mucinous cystadenoma. AB - There are very few benign or malignant diseases which arise in the remnant pancreas after pancreatectomy. Pancreatic carcinoma in the remnant pancreas after pylorus preserving pancreatoduodenectomy (PpPD) for mucinous cystadenoma in a 66 year-old Japanese man is reported in this paper. The patient underwent PpPD for a mucinous cystadenoma in the pancreatic head 39 months prior to the present operation. The surgical margins of the PpPD specimen were free from atypical cells. Follow-up ultrasonography revealed a hypoechoic lesion in the body of the remnant pancreas. Magnetic resonance cholangiopancreatography (MRCP) revealed a stenosis of the main pancreatic duct, with upstream dilatation in the remnant pancreas. Segmental resection of the remnant pancreas, splenectomy, pancreaticojejunostomy and intraoperative radiotherapy were performed under the diagnosis of pancreatic carcinoma of the remnant pancreas. Final histopathological diagnosis was adenocarcinoma of the pancreas. There were no malignant cystic components. The present pancreatic carcinoma was regarded as independent of the previous mucinous cystadenoma. Postoperative radiation therapy and chemotherapy were added. He is doing well 20 months after the second operation although diabetes mellitus has slightly deteriorated. In this communication, we would like to recommend that clinicians should constantly be on guard against the development of pancreatic carcinoma even in the remnant pancreas after pancreatectomy for mucinous cystadenoma. PMID- 9744702 TI - Eikenella corrodens liver abscess complicated by endophthalmitis. AB - A 64-year-old man who presented with sudden, unilateral loss of vision was found to have endophthalmitis associated with a pyogenic liver abscess. The patient was successfully managed with subtotal vitrectomy and percutaneous drainage of the liver abscess. Eikenella corrodens was cultured from the blood and the pus drained from the liver abscess. This is the first reported case of Eikenella corrodens liver abscess complicated by endophthalmitis. PMID- 9744703 TI - Cholestatic hepatitis due to ticlopidine: clinical and histological recovery after drug withdrawal. Case report and review of the literature. AB - A 72-year-old housewife presented with clinical and laboratory signs of acute cholestatic hepatitis. Symptoms had appeared 6 months after she was started on ticlopidine 250 mg/day. Infectious aetiologies were excluded by serology and there was no history of alcohol abuse or use of other drugs. Clinical findings were confirmed by liver biopsy. The drug was discontinued and symptoms gradually subsided. A second biopsy obtained during this phase documented complete resolution of the hepatic damage. A review of the literature shows that the late onset of hepatic toxicity in this case is unique and this is the first report to include histological documentation during the acute phase and after recovery. PMID- 9744704 TI - Colonic pseudo-obstruction due to beta2-microglobulin amyloidosis after long-term haemodialysis. AB - Beta2-microglobulin (A beta2M) amyloidosis is a relatively new secondary amyloidosis associated with renal failure and haemo- and peritoneal dialysis. Although beta2-microglobulin depositions are systemic, clinical manifestations are limited to articular and para-articular disease in most cases. A very limited number of patients have been reported with extra-articular manifestations including those of the gastrointestinal tract. We report on a patient who was treated with haemodialysis for 23 years and developed colonic pseudo-obstruction due to A beta2M amyloidosis. PMID- 9744705 TI - Nodular regenerative hyperplasia of the liver associated with a Factor V Leiden mutation. PMID- 9744706 TI - Improving the utilization of clinical laboratory tests. AB - Reimbursement policies for health care services are greatly diminishing in the U.S. and Western Europe. Hence, there is an increasing need for doctors and other care givers to reduce costs without compromising the quality of the care being delivered. The clinical laboratory is viewed as an area of high costs where significant reductions have been targeted. Efficient utilization of laboratory services can be achieved by elimination of the general health panel, removal of old tests or those that provide redundant information, a reduction in the use of standing orders, more judicious use of drug assays, acceptance of clinical practice guidelines, and use of reflex testing algorithms. New technologies such as DNA probes can substantially improve diagnostic efficiency. Point-of-care testing devices which have higher costs than incremental central laboratory expenses should only be used if they reduce overall operating expenses. Implementation of expert systems can make remaining tests more effective. Doctors and laboratorians must collaborate to achieve more efficient utilization practices. PMID- 9744707 TI - Challenges in evaluating primary health care for teenagers. Research Sub Committee of the Adolescent Working Party of the Royal College of General Practitioners, London, UK. AB - This paper concerns the evaluation of health care for teenagers and examines the role of primary care and its interaction with the teenage users of this service. It recognizes that the majority of health care for teenagers takes place within general practice. The challenge posed is to identify and put in place suitable evaluation tools. There are government targets to improve the health of teenagers by reducing teenage pregnancy, drug use, smoking rates and suicides. It is an assumption of this paper that improvements in experiences of primary care will lead to improvements in more population-based outcomes of care, although this link needs investigation. The paper shows that there are few measures of generic outcome which are available for use in experiments to assess teenage health care as a baseline now. This has implications for conducting future research projects. Such measures are important and it is a necessary feature of research into teenage health that these measures are devised, tested and validated as a priority. PMID- 9744708 TI - Improving clinical outcome in bacteremia. AB - Bacteremia is associated with significant morbidity and mortality. There is wide variation in morbidity and mortality rates according to organism and predisposing conditions. Additionally, prompt administration of appropriate antimicrobial agents is associated with a decrease in mortality. Unfortunately, many bacteremic patients receive inappropriate or no antibiotics. Infectious disease consultation can decrease the number of patients receiving inappropriate initial therapy. 'Quality standard for the treatment of bacteremia' (Gross et al., 1994, Infection Control and Hospital Epidemiology 15, 189-192) is a consensus paper; its purpose is to 'improve the treatment of hospitalized patients with documented bacteremia by ensuring that they receive an antibiotic appropriate in light of the blood culture susceptibility of the pathogen isolated.' A programme to assess the treatment of bacteremia can improve the quality of care with a modest commitment of additional resources. Many of the activities could be performed by a pharmacist, infection control practitioner, or pathologist. However, physician-to physician communications are most likely to be successful. This programme should be considered a component of a hospital's quality-improvement programme; either the hospital quality assurance or infection control committee could be responsible for the programme. We encourage adoption of the standard, and recommend prospective monitoring to include the choice of empiric antimicrobial agents. PMID- 9744709 TI - Methodological challenges to prospective study of an innovation: interregional nursing care management of cardiovascular patients. AB - This paper discusses the methodological challenges of conducting an interregional intersystem study in a highly competitive health care environment. Nurses from a three-hospital rural health consortium and an urban tertiary medical centre collaborated in a two-phase study (a) to describe current interregional cardiovascular health care process (phase I) and (b) to test an interregional nurse-coordinated cardiovascular health care model (phase II). Phase I was a 1 year exploratory descriptive retrospective study involving patient interviews and chart reviews. Findings from the phase I study suggested a pattern of patient health problems occurring weeks and months following successful tertiary centre interventions. Phase II was a quasi-experimental prospective study with 90 intervention subjects and 64 comparison group subjects who completed pencil and paper survey tools over a 12-month period following discharge from the tertiary centre. Nurse and doctor providers completed a satisfaction survey twice, at an interval of 1 year. Concurrent data collection via telephone calls (comparison group) and a retrospective record review (intervention group) provided data on cost, resource use and system efficiency. Initial results of phase II are presented, together with tests of significance for hypotheses related to interregional nurse care management and patient outcomes, patient satisfaction, cost/system efficiency, and provider satisfaction. Methodological considerations and recommendations related to phases I and II of this interregional collaborative cardiovascular project are presented and discussed. PMID- 9744710 TI - Action research--a model for introducing standardized health assessment in general practice: an exploratory study. PMID- 9744711 TI - Riposte to Guest Commentaries on 'Problems associated with randomized controlled clinical trials in breast cancer. AB - This paper addresses the objections of Professor M. Baum and Mr W. J. Cunliffe to my thesis that the randomized controlled clinical trial is a poor tool for the investigation of the treatment of breast cancer, argued in a discussion paper entitled 'Problems associated with randomized controlled clinical trials in breast cancer' (A.E. Johnson, 1998, Journal of Evaluation in Clinical Practice 4, 119-126). The objections range from those that have a philosophical basis, through those founded on differing concepts of the classification of primary tumours and the nature of the metastatic tumour, to those that question the reliability and usefulness of the clinical evaluation of response to treatment in terms of histological grade and rate of tumour shrinkage. An alternative approach to research through primary systemic therapy with selection of treatment according to predicted tumour behaviour was severely criticized, both on the preceding grounds and because it was assumed that the alternative to randomization is management by anecdote. These objections are examined and evidence in support of reliable and useful clinical measurement of response is presented in some detail. The problems associated with randomization as a technique for the evaluation of treatments, when the intrinsic variability of tumours is very large without the intervention of treatment, remain unsolved. PMID- 9744712 TI - A preliminary investigation of patient and carer expectations of their general practitioner in longer-term stroke care. PMID- 9744713 TI - Factors affecting general practice patient response rates to a postal survey of health status in England: a comparative analysis of three disease groups. PMID- 9744714 TI - Primeval health economics in Britain: a personal retrospect of the pre-HESG (Health Economists' Study Group) period. AB - There is a danger that the history of health economics in Britain comes to be regarded as roughly co-terminous with the history of the Health Economists' Study Group (HESG). As one of the founders of that Group, I would take some pride from that, if it were true. But it is not. Just as primitive human societies existed before recorded history, so there was primeval health economics in Britain prior to 1972. There is probably more of this primeval health economics than even I know about, but as one of the ancient relics of that period I have been offered the opportunity to reminisce about what I saw during those dark ages! When one reaches the advanced age of 70, there is no escaping the fact that your past is bound to be more extensive (and probably more enjoyable) than your future, which is why the old enjoy looking back more than they enjoy looking forward! I am no exception. Hence this essay, which may either be seen as a rather self-indulgent bout of nostalgia concerning the early days of health economics in Britain, or as an archaeological enterprise, exhibiting, for all to wonder at, the treasures to be found at carefully selected ancient (i.e., pre-HESG) sites in Britain. Either way, my purpose is to suggest that most of the fundamental issues with which health economists have grappled in the last 25 years had already been identified and addressed in a careful way during the decade preceding the formation of the HESG. PMID- 9744715 TI - From private club to professional network: an economic history of the Health Economists' Study Group, 1972-1997. AB - HESG was founded in 1972 as part of a conscious effort to establish health economics as an identifiable sub-discipline. It is debatable whether the growth of health economics was demand-led or supplier-driven, but in either case the existence of a HESG played a vital role. HESG was founded as a private club, in the tradition of English gentlemen's clubs, designed to provide a forum for debate and an invisible, supportive faculty for health economists dispersed between different organisations throughout the UK. It was given impetus by public economists at the University of York, who were effectively academic entrepreneurs, motivated in part by private gain, but by their actions overcoming the free-rider problem that might otherwise have retarded the development of health economics. Over the course of its first 25 years, HESG has changed and its membership has grown and altered in composition - over this period, HESG has evolved from a private club to a professional network. It has made a vital contribution to the existence and form of health economics as a subdiscipline in the United Kingdom, and has in turn itself been influenced by the subdiscipline. As a subdiscipline, UK health economics in the 1990s generally draws on a small body of economic theory and is practised by a distinct, identifiable group of economists. This paper was commissioned by HESG, as a history of the organisation. It also analyses the foundation and evolution of HESG as an institutional arrangement designed to overcome a collective action problem. PMID- 9744716 TI - The impact of health economics on health policy in England, and the impact of health policy on health economics, 1972-1997. AB - This paper contains a review of the impact of health economics on health policy in England during the past 25 years. Some health economists have expressed disappointment with the scale of the impact that health economics has had on policy but the record set out below suggests that there is modest cause for celebration. That is not to say that there is cause for complacency. There is still a long way to go before all important health policies are based on sound economics reasoning and evidence. The paper begins with some definitions and background; it covers nine areas of health policy, and health policy making, where past impacts of health economics have been postulated; it covers briefly the reciprocal impact of health policy on health economics; and it concludes with a discussion about the findings. PMID- 9744717 TI - Where are we now in British health economics? AB - Health economics took off in 1970 or thereabouts, just after the take-off date for the economics of education. Although early health economics made use of human capital theory as did the economics of education, it soon took a different route inspired by Arrow's work on medical insurance. The economics of education failed to live up to its promising start in the 1960s and gradually ran out of steam. The economics of health, however, has made steady theoretical and empirical progress since 1970, principally in coming to grips with the implications of supplier-induced demand and the difficulties of evaluating health care outcomes. Some of the best work on British health economics has been in the area of normative welfare economics, defining more precisely what is meant by equity in the delivery of health care and measuring the degree of success in achieving equity. Recent efforts to reform the NHS by the introduction of 'quasi markets' have improved the quantity and quality of health care in Britain. In short, British health economics has been characterised by the use of Pigovian piecemeal rather than Paretian global welfare economics, retaining a distinctive style that sets it apart from American health economics. PMID- 9744718 TI - US and UK health economics: two disciplines separated by a common language? AB - Victor Fuchs recently conducted two survey questionnaires of American health economists, showing substantial consensus among them on positive questions and much less consensus on policy questions. I attempted to replicate Fuch's surveys for members of the HESG. I dropped some items that were specific to an American context and added some new questions. Overall there was less agreement on positive questions and more on policy questions than among the US economists. Alan Williams' 1985 article 'Economics of Coronary Artery Bypass Grafting' was deemed the most influential by a British author on health economics as a discipline and British health policy. PMID- 9744719 TI - How do UK gynaecologists manage endometrial carcinoma? A national survey. AB - OBJECTIVE: The objective of this study was to examine the management of endometrial carcinoma in the United Kingdom (UK). METHODS: Consultant gynaecologists were sent an anonymous postal questionnaire requesting information on investigation and management of endometrial carcinoma. RESULTS: Completed questionnaires were received from 701 respondents (51% response). Most responders were general gynaecologists who saw less than five new cases of endometrial cancer annually; 98% of respondents preferred surgical procedure for endometrial carcinoma was total abdominal hysterectomy and bilateral salpingo-oophorectomy. There was a wide variation in diagnostic investigations and radiotherapy practice. PMID- 9744721 TI - The natural history of CIN I lesions. AB - The published literature indicates 11% of CIN I lesions on average progress to a higher grade dysplasia and the remainder either regress or persist. Reliable markers of disease outcome are yet to be identified. A longitudinal study of 342 women referred for colposcopic examination of a CIN I detected by a screening Pap test, and classified by the colposcopic impression and Pap test at that exam as 4 cms; 3) V.B.P. scheme: cis-platinum 50 mg/m2/day 1; vincristine 1 mg/m2/day 1; bleomycin 25 mg/m2/days 1-2-3 (3 courses with 10 days interval); 4) Clinical and sonographic tumor response evaluation following U.I.C.C. response criteria; 5) Radical hysterectomy; 6) Pathological risk factor evaluation; 7) ACH with P.M.C. (cis-platinum 50 mg/m2, methotrexate 30 mg/m2, cyclophosphamide 500 mg/m2) 3 courses every 21 days; 8) Comparison and location of recurrences with a neoadjuvant group (NCH + RH + RT to whole pelvis), and with a control group treated with conventional radiotherapy were done. For statistical analysis the Chi-Square was used and D.F.S. and overall survival (O.S.) were calculated according to the Kaplan Meier and Log Rank Test. RESULTS: After a median follow-up of 75 months (range 42-108), O. S. for stage Ib was 88%, Stage IIb 78%, and 50% for IIIb. The recurrences were 12% (4/34) for Stage Ib (3 local and 1 distant); 28% for IIb (5/18) (4 pelvic and 1 distant); 50% (2/4) for IIIb (2 pelvic recurrences). When residual tumor volume was < 2 cm in the surgical specimen (n=39) there were 4 recurrences (3 pelvic and 1 distant), and 7 (6 pelvic and 1 distant) for tumors > 2 cm. (p<0.01 for pelvic recurrences). For the stage Ib with residual tumor <2 cm (n=14) there were no pelvic recurrences and only 1 distant. Comparing for Stage Ib with previous tumor volume >4 cm of the ACH Group (n=17) with a classical NCH (n=51) and control (n=51) groups, there were observed 2 (11.7%) pelvic and 1 (5,8%) distant relapses for the 1st Group, 3 (5.9%) pelvic and 3 (5.9%) distant relapses for the 2nd, and 11 (21.6%) pelvic and 5 (9.8%) distant relapses for the 3rd Group. From the comparison of locally advanced tumors (Stages IIb + IIIb) of ACH group (n=22), with a Stage IIIb surgically removed of classical NCH group (n=38) and with a control group of conventional RT (n=51), there were observed 6 (27%) pelvic and 1 (4.5%) distant relapses for the 1st Group, 4 (11%) pelvic and 7 (18.4%) distant relapses for the 2nd, and 31 (60.7%) pelvic and 5 (9.8%) distant for the 3rd one. CONCLUSION: ACH after NCH + RH could be used for stage Ib with tumor volume > 4 cm, with complete clinical response or residual tumor < 2 cm. The results of this group of tumors suggest the importance of going on phase III trials comparing NCH+RH alone vs. NCH+RH+ACH. ACH could also be used to try to obtain better control of distant metastases in Stages IIb and IIIb. In these cases radiotherapy to the whole pelvis must not be excluded. PMID- 9744729 TI - A pilot study on early post-operative morbidity and technique of inguinal node dissection in vulval carcinoma. AB - Inguinal lymphadenectomy is part of the management plan for most cases of vulval carcinoma. The surgical techniques have been modified over the years resulting in less destructive operations. Even so, inguinal lymphadenectomy continues to pose difficulties particularly relating to wound breakdown and lymphocyst formation. Many different methods are described, though none have undergone any comparative assessment regarding morbidity. This small study compares two methods performed on the same patients. The radical procedure included excision of the fascia lata and exposure of the femoral vessels and nerve. The anatomically-directed method was more conservative with surgery directed at removing the nodes as described in anatomical textbooks. Both methods resulted in equal lymph node retrieval, though the subjective short-term morbidity was reduced with the more conservative surgery. This approach did not result in any detrimental outcomes regarding relapse disease, though a randomised trial is required to corroborate these findings. PMID- 9744730 TI - Expression of the cell adhesion molecules ICAM-1 and VCAM-1 in the cytosol of breast cancer tissue, benign breast tissue and corresponding sera. AB - OBJECTIVE: Cellular adhesion molecules ICAM-1 and VCAM-1 have been implicated in tumor progression and metastasis. As the sequential interaction of neoplastic cells with the endothelium of tumor neovascularisation is believed to be essential for tumor metastasizing processes, we analysed the concentration of ICAM-1 and VCAM-1 in the cytosol of patients with human breast cancers and their corresponding sera. We compared the obtained values with established prognostic parameters for breast cancer. Benign breast tissues were also analyzed. PATIENTS AND METHODS: Levels of ICAM-1 and VCAM-1 of 62 patients with invasive breast cancer and 17 patients with benign breast tissue were measured using commercially available sandwich enzyme-linked immunoassays with monoclonal antibodies. To establish a reference and control group, levels of ICAM-1 and VCAM-1 were measured in the sera of 66 women without breast tumors. RESULTS: The mean cytosol concentration of ICAM-1 and VCAM-1 was significantly higher in the breast cancer specimens than in the tissue of patients with benign breast diseases. This could be found not only in the tumor cytosol but also in the corresponding sera of the patients. No correlations between the ICAM-1 and VCAM-1 expressions and established prognostic parameters could be observed. CONCLUSIONS: Our findings suggest that malignant breast cancer cells could induce neovascularisation with subsequent high expressions of ICAM-1 and VCAM-1. These upregulations of adhesion molecules might contribute to changes in invasive phenotypes by promoting endothelial cell adhesion and angiogenesis, as well as being responsible for the recognition of tumor cells by the human immune system. Prognostic relevance for the development of breast cancer could not be established. PMID- 9744731 TI - Primary carcinoma of the fallopian tube. PMID- 9744733 TI - Pseudo-Meigs' syndrome caused by paraovarian fibroma. AB - Meigs' syndrome includes an ovarian tumor, usually fibroma, associated with hydrothorax and ascites. It is accepted that uterine tumors, like fibromas, can also be associated with ascites and hydrothorax, but this is extremely rare. The mechanism of formation of peritoneal and pleural effusion is not well documented. The most likely pathogenesis ascribes the fluid formation to the filtration of interstitial fluid in the peritoneal through the tumor capsule, and the diffusion to the pleural space through the diaphragm lymphatic vessels at the foramen of Bochdalek. Paraovarian fibromas are also extremely rare neoplasms, probably of paramesonephric origin. It has been hypothesised that they can develop by proliferation of connective tissue cells around the Wolfian remnants. In this article, probably for the first time, a case of paraovarian fibroma with ascites and hydrothorax is presented. PMID- 9744732 TI - Vulvar metastasis from breast carcinoma: a case report and review of the literature. PMID- 9744734 TI - Benign epithelial ovarian tumors. AB - Two hundred and twelve patients with benign epithelial ovarian tumors managed consecutively at Hacettepe University Hospital between 1974-1994 were analyzed retrospectively. Benign epithelial tumors constituted 9.5% (212/2216) of all ovarian tumors and 28.5% (212/743) of the primary epithelial ovarian tumors during the study period. Of the patients with benign epithelial tumors, 104 (49.0%) had serous, 94 (44.3%) had mucinous and the remaining 14 (6.7%) had Brenner tumors. The surgical procedures varied from cystectomy to total abdominal hysterectomy according to the age of the patient. Bilaterality was encountered in 12.8% of serous and 10.6% of mucinous tumors. Ovarian carcinoma occurred synchronously in two patients. In two patients, serous cystadenoma was detected in the preserved ovaries following 3 and 7 years of initial conservative surgery, respectively. Serous tumors were relatively more common constituting approximately one half of the cases. Most of the patients presented with an abdominopelvic mass. PMID- 9744735 TI - Study on interaction of proteins isolated from epithelial cervical tissue with glucocorticoid-response element of HPV16. AB - Genital HPV16 has tropism to cervical epithelial tissue which is highly steroid hormone dependent. Protein/DNA interaction with the fragment of viral DNA containing the glucocorticoid-response element (GRE/PRE) did not indicate the proteins which can bind to the GRE element in normal epithelial cervical tissue. NFI is one of the cellular factors which can participate in the transcription of viral genes in study tissue. PMID- 9744736 TI - The role of mammography in the diagnostic approach of breast hamartomas. AB - PURPOSE: Hamartomas are rare benign breast tumors which surfaced with the entrance of mammography in the diagnostic algorithm of breast lesions. In the present study the mammography findings of breast hamartomas were analyzed in relation to the histologic confirmation of the diagnosis. PATIENTS AND METHODS: During the seven-year period between January 1990 and January 1997 we studied 45 patients, aged from 32 to 66 years, with a clinical and/or mammographic finding of a breast lump, highly suspicious of hamartoma. In 29 (64.4%) the tumor was found in the left breast and in the remaining 16 (35.6%) in the right breast. RESULTS: The preoperative mammography of 41 patients revealed a nodular opaque mass of a nonhomogeneous composition and only four had the typical hamartoma picture. The tumor diameter ranged between 0.7-6.5 cm. All tumors were excised and the histological reports confirmed the diagnosis of hamartoma. CONCLUSION: Mammography helps in the diagnosis of hamartomas. Nevertheless, the final diagnosis must be based on the histological analysis of the tumor. PMID- 9744737 TI - Pregnancy-associated breast cancer. AB - Pregnancy-associated breast cancer, which is defined as all breast cancer diagnosed during pregnancy or within the following year, is a relatively rare finding. Due to the particular difficulties in the diagnosis of breast cancer during this period, pregnant women tend to present more advanced disease at diagnosis. Four cases of pregnancy-associated breast cancer referred to our Institute are described as a contribution to the knowledge of this disease. PMID- 9744739 TI - Case report: endometrial cancer implanting in the laparotomy scar. AB - A 58-year-old woman underwent abdominal hysterectomy and bilateral salpingo oophorectomy for stage Ib, grade 2 endometrial adenocarcinoma followed by external pelvic irradiation. Five years later she presented with a 7 cm solitary infraumbilical incisional tumor recurrence that was resected. Histology of the tumor implant was similar to that of the primary cancer. The patient was then started on progestin therapy with no evidence of recurrence for four years. To our knowledge this is the fourth reported case of endometrial cancer implanting in an abdominal scar. PMID- 9744738 TI - Toxicity of chemotherapeutical protocols in the treatment of uterine sarcomas (Vincristine, actinomycin D, Cyclophosphamide VAC versus ifosfamide). AB - OBJECTIVE: Uterine sarcomas are rare tumors which account for 1% of all genital tract malignancies. They have a poor prognosis with an overall survival of under 50% at 2 years. The benefit of chemotherapy is unclear and different chemotherapy protocols are used for the treatment of uterine sarcomas. But there is little experience about their toxicity because of the limited case series. So we compared VAC protocol and ifosfamiide for toxic effects. MATERIAL AND METHOD: We reviewed 13 cases which were diagnosed as uterine sarcomas and treated with surgery plus chemotherapy at The Department of Obstetrics and Gynecology, Akdeniz University School of Medicine from 1990 to 1995. Data were obtained from patient files. RESULTS: Mean age was 55.7 (range 38-70), 7 (53.8%) patients had malignant mixed mullerian tumors and 6 (46.1) had leiomyosarcomas. A total of 32 courses of chemotherapy were given -20 ifosfamide and 12 VAC therapy. Leucopenia, hepatic dysfunction and peripheral neuropathy were more frequent in the VAC group as 75%, 16.6%, versus 30%, 0%, 0%, in the iFosfamide group respectively. However, urothelial toxicity (35%) was more common in the ifosfamide group. CONCLUSION: VAC protocol is more toxic for the liver, hematopoietic and peripheral neurologic system. On the other hand the major toxicity of ifosfamide was on the urinary tract. Ifosfamide may be a good choice with less toxicity than VAC therapy in the treatment of uterine sarcomas. PMID- 9744740 TI - Estrogen and progesterone receptor manipulations in castrated rat uterus models and the clinical adaptation. AB - To investigate the influence of tamoxifen, danazol and triptorelin (a GnRH agonist) on estrogen and progesterone receptors of rat endometrium, 44 castrated Sprague-Dawley rats were divided into four equal groups, and each group received either no treatment or one of the agents. After administration of the agents, estrogen and progesterone receptor levels, detected by immunohistochemical methods, were compared with the controls. Estrogen and progesterone receptors were significantly higher in the tamoxifen group than the controls (p<0.05), but this was not noticed in the triptorelin group (p>0.05). Receptor levels were higher in the danazol group than the controls, but it was significant only in the estrogen receptors. Among the 3 groups, receptor levels were higher than in the control group. There was not any correlation among the estrogen and progesterone receptor levels in all groups. Steroid receptor manipulations can be used in the treatment of gynecologic cancer, but further investigations are needed. PMID- 9744741 TI - Fatal acute tumor lysis syndrome following treatment of vulvar carcinoma: case report. PMID- 9744742 TI - Isolation and purification of immunoglobulins from chicken eggs using thiophilic interaction chromatography. AB - We report on the use of thiophilic interaction chromatography for the purification of IgY from egg yolk. This procedure permits the purification to homogeneity of IgY in a single chromatographic step after ammonium sulfate fractication. This study also compares the use of an improved T-gel which has a higher capacity for immunoglobulin than the original T-gel, having a capacity in excess of 25 mg IgY/ml resin. The recovery from this procedure is close to 100%, providing a simple and efficient means for purifying IgY from egg yolk. We also determined that the amount of specific antibody present in egg yolk from an immunised chicken is around 1% of total IgY. PMID- 9744743 TI - Detection of low-affinity adhesion ligands by linking recombinant cell adhesion molecules in uniform orientation to a fluorescently labelled dextran molecule by means of hexahistidine tagging: the case of multimeric CD40. AB - Cell-cell interactions involve highly polyvalent associations between receptors on adjacent cells. In order to mimic this process, we have prepared a highly polyvalent form of CD40 attached to a dextran backbone. This was accomplished by engineering a hexahistidine tag on the C-terminus of the CD40 and binding, in a uniform orientation, up to 100 molecules of hexahistidine CD40 by metal chelation to a single fluorescently tagged dextran molecule. The advantage of this 'multimeric' CD40 is that it would be expected to bind to any counterstructure with a significantly higher avidity compared to monomeric CD40. The multimeric CD40 bound with high affinity to stably transfected mouse fibroblasts expressing CD40L. The multimeric ligand also bound to the activated T cell clone, D10, but did not bind to resting cells, showing that it bound to the physiological ligand. Using this system, we found no evidence to support the claim [Heath et al., 1993. Cell. Immunol. 152, 468.] that the A20 cells have a counterstructure for CD40, and propose that the high binding of CD40 observed in this study may have been due to an exposed hexahistidine tag on the molecule. This multimeric technology has considerable potential for detecting low-affinity interactions between cell adhesion receptors and ligands. The uniform orientation of the molecules on the dextran is an advantage over previous systems and permits the preparation of heterogeneous, multimeric ligands which more closely mimic the conditions at the cell surface. PMID- 9744744 TI - Kinetic analysis of immunointeractions with covalently immobilized fatty acid binding protein using a grating coupler sensor. AB - Application of a grating coupler sensor (GCS) to the real time investigation of the interaction kinetics of covalently immobilized recombinant bovine heart-type fatty acid-binding protein (H-FABP) and corresponding antibody is described. The immobilization of the antigen is performed by activating the matrix hydroxyl groups with p-toluenesulfonyl chloride (TSC) and afterwards coupling the protein by reaction with its nucleophilic aminogroups. Covalent coupling via TSC permits reproducible measurements of immunointeractions on the same grating coupler sensor chip and complete regeneration after each binding cycle with glycine hydrochloride. We demonstrate the analysis of binding data obtained on a GCS by linearization as well as direct curve fitting using the integrated rate equation for the determination of apparent rate and affinity constants. With both analysis methods we studied H-FABP/monoclonal anti-H-FABP-antibody interactions and obtained an average apparent association rate constant ka = 4.2 X 10(3) M(-1) s( 1) a dissociation rate constant of kd=1.3 X 10(-4) s(-1) and an equilibrium constant of KD=3 X 10(-8) M. PMID- 9744745 TI - Generation of chimeric monoclonal antibodies from mice that carry human immunoglobulin Cgamma1 heavy of Ckappa light chain gene segments. AB - Gene targeting in mouse embryonic stem (ES) cells was used to replace (i) the mouse immunoglobulin heavy chain (IgH) Cgamma2a gene segment (mCgamma2a) with the human Cgamma1 gene segment (hCgamma1), and (ii) the mouse immunoglobulin light chain (IgL) Ckappa gene segment (mC kappa) with its human counterpart (hC kappa). ES cells carrying these gene conversions were used to generate chimeric mice that transmitted the human alleles through the germ line. Mice homozygous for both gene alterations were generated by breeding. Serum from homozygous mutant mice contained comparable amounts of antibodies with chimeric kappa or mouse lambda light chains but only small fractions of basal serum IgG or antibodies elicited against immunizing agents contained chimeric heavy chains. A relative increase in immunogen-specific hCgamma1 antibodies was seen following immunization in combination with the saponin adjuvant QS-21. The effect of this was to shift the IgG1-dominated response to an IgG subclass profile that included significant amounts of IgG2a, IgG2b and IgG3 and chimeric IgG. The amounts of antibody secreted by hybridomas derived from mutant and wild-type mice were similar. Sequencing confirmed correct splicing of hCgamma1 and hCkappa gene segments to mouse J gene segments in hybridoma Ig gene transcripts. In conclusion, IgHhCgamma1/IgLhCkappa double mutant mice provide a useful animal model for deriving humanized antibodies with potential applications in immunotherapy and diagnostics in vivo as well as for investigating hCgamma1 associated functions. PMID- 9744746 TI - Effect of bacterial invasion of macrophages on the outcome of assays to assess bacterium-macrophage interactions. AB - In vitro assays to quantify killing of bacteria by macrophages provide useful insights into host-pathogen relations. In the present study, we used strains of Yersinia enterocolitica and Escherichia coli which varied in their ability to invade mammalian cells to evaluate these assays. The results showed that 30 min and 24 h after incubation with murine bone marrow-derived macrophages, strains of Y. enterocolitica and E. coli which expressed invasin (an outer membrane protein which allows bacteria to penetrate mammalian cells) achieved significantly greater numbers in macrophages than otherwise isogenic bacteria which lacked this protein (P < 0.01). When the 24-h data were corrected for the number of bacteria ingested by macrophages initially, the differences between invasin-positive and negative bacteria were no longer evident (P> 0.2). This study has shown (1) that invasin-mediated penetration of macrophages by bacteria is not associated with enhanced intracellular survival, and (2) that invasion of macrophages by bacteria may influence the interpretation of assays for bactericidal capacity unless allowance is made for the number of bacteria ingested during the early phase of the assay. PMID- 9744747 TI - Competitive reverse transcription polymerase chain reaction for quantifying pre mRNA and mRNA of major acute phase proteins. AB - Competitive reverse transcription polymerase chain reaction (RT-PCR) is an increasingly used method for quantifying RNA. The technique involves co amplification from test RNA with an internal standard using common primers in a single reaction. The standard competes for primers and enzyme and it is therefore referred to as a competitor. A RT-PCR polycompetitor for use in quantifying acute phase serum amyloid A, constitutive serum amyloid A, serum amyloid P component, C reactive protein, apolipoprotein A1, apolipoprotein A2, glyceraldehyde-3 phosphate dehydrogenase and beta-actin mRNAs has been generated and used in quantitative PCR. The polycompetitor was synthesised from ten oligonucleotides of 77-90 bases using primer extension and contains sequences which permit amplification using priming sites that are present in both hnRNA (pre-mRNA) and mRNA. The polycompetitor was cloned into the expression vector pSP64(polyA) and a polycompetitor transcript with a poly(A)-tail sequence at the 3'-end was generated by in vitro transcription. The poly(A)-tail sequence allows the option of performing reverse transcription using oligo(dT) in addition to directed reverse transcription using the specific 3'-reverse PCR primers. cDNA products generated from amplification of the internal polycompetitor standard and endogenous RNA species were separated by capillary electrophoresis and quantified by UV absorbance at 254 nm. Reproducibility was determined to be very high when starting ratios of internal standard and target mRNA are at an approximate equivalence point. Relative standard deviations were less than 5% between independent RT-PCR reactions with the same sample mix of internal standard and total RNA. Applying the method to total RNA samples harvested at various timepoints following cytokine induction of acute phase mRNAs in endothelial cells demonstrated that quantitative readout from the RT-PCR method correlates well with that obtained from Northern-blotting and is at least 100-fold more sensitive. This method will be useful for studying regulation of acute phase proteins in in vitro tissue culture experiments and may also be applied to clinical tissue samples from patients with inflammatory diseases. PMID- 9744748 TI - Enhanced T cell proliferation and increased responder frequency following delivery of antigen to the antigen-presenting cell; B cell dependency and use in detection of autoreactive T cells. AB - Reported frequencies of peripheral blood autoantigen-specific cells in autoimmune diseases are typically low, which could be due to true scarcity or to limitations of in vitro assays. In the present study, antigens were targeted to the antigen presenting cell (APC) to enhance T cell proliferation, using an antigen delivery system (ADS), consisting of biotinylated anti-IgG, streptavidin and biotinylated antigen. This was able to bind B cells and monocytes and was internalized within 24 hours. T cell proliferation to tetanus toxoid was at least doubled using the ADS compared to conventional assay with antigen in simple solution. To evaluate the ADS in an autoimmune disease, we determined T cell responses to the insulin dependent diabetes mellitus (IDDM)-associated autoantigen IA-2ic in patients with recent-onset IDDM. When IA-2ic was available conventionally in solution, proliferation was poor, but significantly higher in IDDM patients than control subjects. However, the ADS significantly enhanced proliferation by a mean 3-fold for all subjects, while maintaining the significant difference between IDDM patients and healthy controls. Increases in T cell proliferation via the ADS were due to the recruitment of approximately 3 times the number of CD4 + T cells stimulated in conventional assays. B cell depletion abolished enhancement suggesting that the ADS operates through recruitment of B cells as APCs. This flexible modification of the T cell assay offers greatly enhanced sensitivity for determining the frequency of antigen and autoantigen-reactive T cells. PMID- 9744749 TI - A europium fluoroimmunoassay for measuring peptide binding to MHC class I molecules. AB - A fluoroimmunoassay employing europium-streptavidin and time-resolved fluorimetry was used to measure binding of biotin-labeled peptides to H-2Dk molecules. A fluorescein-labeled octameric peptide from the middle T (MT) protein of mouse polyoma virus was identified that specifically binds to purified Dk using a previously-established assay based on size exclusion chromatography. A europium immunoassay was adapted to measure binding of a biotin-derivative of this peptide to purified Dk. The assay was found to be sensitive and highly specific. Binding was optimal at pH 5.0 and 24-27 degrees C, and it was not enhanced in the presence of additional beta2-microglobulin (beta2m). Excellent results were also obtained in experiments with fixed cells that express Dk. This assay is expected to be useful for high volume, routine analysis of peptide binding to MHC class I molecules. PMID- 9744750 TI - Improved IL-2 detection for determination of helper T lymphocyte precursor frequency in limiting dilution assay. AB - In the context of allogeneic bone marrow transplantation, an accurate estimate of the risk of developing graft-versus-host disease (GVHD) is of major interest. The pre-transplant frequency of donor's helper T-lymphocyte precursors (HTLp) directed against host's antigens may be helpful in predicting this risk. This technique relies on an indirect measurement of interleukin-2 (IL-2) secreted by the HTLp, as assessed by the proliferation of an IL-2 dependent cell line. Many authors use the murine CTLL-2 cell line in this assay, but these cells do not respond to the presence of minute amounts of IL-2 in the culture medium, and thus do not discriminate between the absence or the presence of very low levels of IL 2. We therefore decided to compare CTLL-2 with another IL-2 dependent cell line, the murine A9.12 cell line. A comparison was made using serial dilutions of recombinant human IL-2, limiting dilutions of baby hamster kidney (BHK) cells transfected with human IL-2 gene and in the context of clinical tests performed for the detection of pre-transplant HTLp. Both the sensitivity and reliability of the tests were better using A9.12. We conclude that the A9.12 cell line might be a more suitable tool for pre-transplant HTLp determinations before allogeneic bone marrow transplantation or whenever low IL-2 levels are to be measured. PMID- 9744751 TI - Immunochemical approaches to AGE-structures: characterization of anti-AGE antibodies. AB - Recent immunological approaches have greatly helped broaden our understanding of the biomedical significance of advanced glycation end products (AGEs) in aging and age-enhanced disease processes. Recently, Nepsilon-(carboxymethyl) lysine (CML), one of the glycoxidation products of AGEs, was demonstrated to be a major immunological epitope among AGEs. In the subsequent study, we characterized 13 different polyclonal anti-AGE antibodies and showed that these antibodies could be classified into three groups (Groups I, II and III). Group I was specific for CML and both Group II and Group III were specific for other epitopes (non-CML). Time-course study suggested that the epitope of Group II was formed earlier than that of Group III. In the present study, we prepared two monoclonal anti-AGE antibodies (2A2 and 3A3) whose epitope structures appeared to be closely related to Group III and Group II, respectively. The result indicates that AGE-proteins express at least two major non-CML epitopes. PMID- 9744752 TI - Optimized conditions for gene transfection into the human eosinophilic cell line EoL-1 by electroporation. AB - Eosinophils are emerging as an increasingly important cell in the immunoregulatory network of normal and pathological processes. No studies has yet described optimized experimental strategies to transfect DNA into human eosinophils. Using a frequently employed in vitro model of human eosinophil, the EoL-1 cells, we now described the optimal transfection of DNA into these cells by electroporation. Our results indicate that electroporation can efficiently and reproducibly transfect DNA into EoL-1 cells. Optimal electroporation conditions consist of the use of 1 X RPMI medium 1640 with 10% FBS, voltage setting at 275 V, 1150 microF capacitance, 40 mg of DNA and 4.0 X 10(7) cells/ml per electroporation in a total volume of 0.5 ml in 0.4 cm gap cuvettes. These conditions may be a useful protocol for transfecting eosinophil cell lines. PMID- 9744753 TI - High-resolution analysis of T-cell receptor beta-chain repertoires using DNA heteroduplex tracking: generally stable, clonal CD8+ expansions in all healthy young adults. AB - The accurate measurement of T-cell receptor (TCR) repertoire changes requires the analysis of a representative sampling of complex T-cell populations. The number and frequency of clonally expanded TCR beta-chain transcripts bearing distinct CDR3 sequences were accurately determined using a simple DNA heteroduplex tracking assay. This method allowed major and minor clonal expansions (> or = 1% of a Vbeta subfamily's transcripts) to be rapidly and reproducibly quantified. Oligoclonal CD8 + cell expansions were detected in all young adults tested, while CD4 + cells generally expressed more polyclonal beta-chain repertoires. The same pattern of CD8 + cells oligoclonality and CD4 + cells polyclonality was observed in asymptomatic HIV-1 infected individuals with high CD4 + cell counts. CD8 + CD45RA + and CD8 + CD45RO + cell fractions both displayed oligoclonal, although distinct, TCR beta chain repertoires while CD8 + cells from umbilical cord blood were generally polyclonal. Oligoclonal CD8 + cell repertoires from young adults were generally stable over a period of weeks, although minor, transient, clonal expansions could also be detected in the absence of symptomatic infections. DNA heteroduplex tracking analysis provided a higher level of sensitivity for the detection of TCR beta chain transcript expansions than CDR3 length (spectrotyping/immunoscope) analysis. DNA heteroduplex tracking of TCR beta-chain transcripts is therefore a simple and sensitive method for assessing the level of clonality and for measuring changes in the TCR beta chain repertoire of different T-cell populations. PMID- 9744754 TI - Quantification of cytokine mRNA in peripheral blood mononuclear cells using branched DNA (bDNA) technology. AB - Changes in the patterns of cytokine expression are thought to be of central importance in human infectious and inflammatory diseases. As such, there is a need for precise, reproducible assays for quantification of cytokine mRNA that are amenable to routine use in a clinical setting. In this report, we describe the design and performance of a branched DNA (bDNA) assay for the direct quantification of multiple cytokine mRNA levels in peripheral blood mononuclear cells (PBMCs). Oligonucleotide target probe sets were designed for several human cytokines, including TNFalpha, IL-2, IL-4, IL-6, IL-10, and IFNgamma. The bDNA assay yielded highly reproducible quantification of cytokine mRNAs, exhibited a broad linear dynamic range of over 3-log10, and showed a sensitivity sufficient to measure at least 3000 molecules. The potential clinical utility of the bDNA assay was explored by measuring cytokine mRNA levels in PBMCs from healthy and immunocompromised individuals. Cytokine expression levels in PBMCs from healthy blood donors were found to remain relatively stable over a one-month period of time. Elevated levels of IFNgamma mRNA were detected in PBMCs from HIV-1 seropositive individuals, but no differences in mean levels of TNFalpha or IL-6 mRNA were detected between seropositive and seronegative individuals. By providing a reproducible method for quantification of low abundance transcripts in clinical specimens, the bDNA assay may be useful for studies addressing the role of cytokine expression in disease. PMID- 9744755 TI - The usefulness of Diffusion-In-Gel-ELISA in clinical practice as illustrated by a Campylobacter jejuni outbreak. AB - In this study, the DIG-ELISA (Diffusion-In-Gel Enzyme Linked Immunosorbent Assay) is presented as a tool for the determination of antibodies with improved quantitation over other solid phase assays. The method combines the diffusion of antibodies in agar with the EIA concept in Petri dishes. Diffusion of native, undiluted sera results in (1) a concentration gradient in the effort to reach equilibrium, i.e., an end point titration; (2) a separation of serum components of different sizes; (3) unlimited possibilities for migrating antibodies to bind antigenic epitopes because of the constant antigen excess. Low affinity antibodies can bind divalently and are more readily detected; and (4) elimination of dilution errors. The combination of undiluted sera and Petri dishes as solid phase also permits a large number of samples to be tested with less effort and simplifies the practical handling of EIAs, including easy coating and washing procedures, reuse of antigen and quantitation by zone areas without instrumentation. Plates can be stored for months and are available for re examination, demonstration and transport. The whole procedure can be conducted in a closed system, i.e., when testing highly contaminated samples. The usefulness of the procedure is demonstrated by a food-borne outbreak of Campylobacter jejuni. The outbreak involved 86 persons of whom 20% were culture positive in contrast to a seropositivity of 74% with DIG-ELISA. A diffusion time of 24 h was used for diagnostic purposes. Extended diffusion times of 48 h and 72 h were utilized when consecutive series of sera showed identical values after 24 h, indicating high antibody content that resulted in a peak serum (end point). For infectious diseases with a rapid course, this assay could be used as an acute test. With the diffusion step prepared in advance, DIG-ELISA is a ready-to-use test. When frequent sampling of sera is performed in the very early phase of the disease, DIG-ELISA reveals that all Ig-classes can be present at high titer and the diagnostic potential of the immune response is better utilized. The DIG-ELISA has the methodological flexibility and the physical qualities to be an effective, inexpensive technique for quantitation of antibodies at any laboratory. PMID- 9744756 TI - Expressed luciferase viability assay (ELVA) for the measurement of cell growth and viability. AB - An expressed luciferase viability assay (ELVA) has been developed for cell viability and cell number based on detecting the expression of luciferase transfected into the cells. Stable transfectants were produced that expressed luciferase constitutively. Like many endogenous enzymes, luciferase is rapidly degraded following cell death, so that the enzyme can be used as a measure of cell viability. A modified luciferase assay was used in which the reagents were added directly to the cells in a microplate. The main advantages compared to other cell viability assays are the wide dynamic range, high sensitivity, low background, and the absence of any requirement to wash or harvest the cells. Stable transfectants of three factor-dependent cell lines (B13, Ba/F3 and CTLL) were produced and used in cytokine assays. Three strategies of selection after electroporation were tested: (1) using a plasmid containing both the genes encoding firefly luciferase and a selectable marker (neo), (2) cotransfection of a plasmid containing luciferase and a plasmid containing a selectable marker (puromycin resistance), and (3) cotransfection of a plasmid containing luciferase and a plasmid containing the human IL-5Ralpha-chain, and selecting in IL-5. This latter strategy produces an IL-5 responsive cell line expressing luciferase in a single step without the need for antibiotic selection. PMID- 9744757 TI - Cross-linked filamentous phage as an affinity matrix. AB - Filamentous phage can be cross-linked to make a hydrophilic aggregate that is pelleted by low-speed centrifugation. The aggregate is stable at near-neutral pHs, and withstands exposure to the acid buffers (pH down to 2.2) that are often used as eluents in immunoaffinity purification. If a peptide epitope is genetically fused to a coat protein on the virion surface, the aggregate serves as an effective affinity matrix for absorbing and affinity-purifying antibodies that bind the peptide. When the peptide epitope is first obtained in this form by selection from large phage display libraries, this ability to fashion an affinity matrix directly from the selected phage represents a significant streamlining of research and development. PMID- 9744758 TI - Mucosal antibodies can be measured in air-dried samples of saliva and feces. AB - IgA antibodies reflecting airways or intestinal mucosal immune responses can be found in saliva and feces, respectively, and IgG antibodies reflecting serum antibodies can be found in saliva. In this study, antibodies were detected in samples of saliva and feces which had been air-dried at room temperature (+20 degrees C) or +37 degrees C, and stored at these temperatures for up to 6 months. In saliva the antibody levels increased, while the antibodies in feces decreased upon storage. The individual IgA antibody concentrations which were adjusted by using the ratios of specific IgA/total IgA were relatively stable in both saliva and feces, and correlated with corresponding antibody levels in samples which had been stored at -20 degrees C. The results indicate that air-dried saliva and feces can be used for semiquantitative measurements of mucosal antibodies, even after prolonged storage at high temperatures and lack of refrigeration. PMID- 9744759 TI - Transient transfection of primary T helper cells by particle-mediated gene transfer. AB - The study of the molecular basis of normal CD4+ T cell function, such as the control of commitment to the TH1 or TH2 phenotypes has been difficult due to the resistance of these cells to transfection by conventional methods. We used antibodies specific to T cell surface molecules to immobilize these cells and optimized conditions for transiently transfecting them by means of particle mediated gene transfer. Using this technique, a construct encompassing - 577 to +1 of the IL-4 promoter allowed transcription of a luciferase reporter gene in recently-differentiated TH2 cells stimulated by anti-CD3, consistent with regulation of endogenous IL-4 gene expression. PMID- 9744761 TI - Cytokine production in human mixed leukocyte reactions performed in serum-free media. AB - The mixed leukocyte reaction (MLR) is an in vitro test commonly performed in a serum-containing medium (SCM), and used to study allorecognition and cellular immunity accompanied by cytokine release. We investigated the possibility of performing the MLR test in serum-free media (SFM) by comparing human leukocyte proliferation and cytokine release in MLRs performed in SFM and SCM. Of the four SFM tested, only Biotarget- was as effective as SCM in supporting leukocyte proliferation and IL-2 secretion. Both phenomena were observed only in allogeneic combinations. The levels of IL-1, IL-6, and TNFalpha in allogeneic MLR combinations in SFM were half those in SCM cultures; the kinetics of their release were the same. With the exception of IL-2, a high degree of spontaneous release of the other three cytokines analyzed was observed in responder cells, in irradiated stimulator cells, and in autologous combinations cultured in both SCM and SFM. It appears that unlike IL-2, the cytokines IL-1, IL-6, and TNFalpha are nonspecifically produced in MLR and cannot serve as sensitive indices of HLA disparity. PMID- 9744760 TI - Generation, characterization and utilization of anti-human growth hormone 1-43, (hGH1-43), monoclonal antibodies in an ELISA. AB - The major isoform of hGH is a polypeptide of 191 amino acids. Human GH1-43 is an amino terminal segment of hGH1-191 which comprises the first 43 amino acids. This peptide is a potent regulator of glucose homeostasis. To facilitate our understanding of the physiological regulation of hGH1-43 an assay to measure its levels in biological fluids and extracts is needed. This communication describes the development of anti-hGH1-43 monoclonal antibodies and their use in the development of an indirect competitive ELISA for the quantification of hGH1-43. Hybridomas were produced by the fusion of FOX-NY myeloma cells with spleen cells taken from a mouse immunized with hGH1-43. The hybridomas were screened for production of antibodies to hGH1-43 by antibody capture ELISA. Hybridomas which produced antibodies reactive to hGH1-43 were cloned by limiting dilution. Three monoclonal hybridomas, CCL-1, CCL-2, and CCL-3 were subsequently obtained. These hybridomas secreted antibodies that were highly reactive towards hGH1-43 but minimally reactive towards hGH1-191. The isotypes of the mAbs secreted by CCL-1, CCL-2 and CCL-3 were all IgG1 kappa as shown by isotype specific antibody capture analysis. An indirect competitive ELISA with a detection limit that ranged from 1 to 10 ng/ml was developed using mAbs from monoclonal hybridoma CCL-3. Dose response curves for competing hGH1-191, hPRL, and hPL indicated minimal cross reactivity of mAbs with these hormones and conversely, a high degree of specificity for hGH1-43. Dose-response curves for dilutions of human serum and pituitary extract were parallel to the standard. The availability of a sensitive assay for the measurement of hGH1-43 will help us answer questions regarding the biosynthesis, regulation of secretion, and role of hGH1-43 in the control of glucose homeostasis. PMID- 9744762 TI - Resistance to antimicrobial agents used for animal therapy in pathogenic-, zoonotic- and indicator bacteria isolated from different food animals in Denmark: a baseline study for the Danish Integrated Antimicrobial Resistance Monitoring Programme (DANMAP). AB - This study describes the establishment and first results of a continuous surveillance system of antimicrobial resistance among bacteria isolated from pigs, cattle and broilers in Denmark. The three categories of bacteria tested were: 1) indicator bacteria (Escherichia coli, Enterococcus faecalis, Enterococcus faecium), 2) zoonotic bacteria (Campylobacter coli/jejuni, Salmonella enterica, Yersinia enterocolitica), and 3) animal pathogens (E. coli, Staphylococcus aureus, coagulase-negative staphylococci (CNS), Staphylococcus hyicus, Actinobacillus pleuropneumoniae). A total of 3304 bacterial isolates collected from October 1995 through December 1996 were tested for susceptibility to all major classes of antimicrobial agents used for therapy in Denmark. Bacterial species intrinsically resistant to an antimicrobial were not tested towards that antimicrobial. Acquired resistance to all antimicrobials was found. The occurrence of resistance varied by animal origin and bacterial species. In general, resistance was observed more frequently among isolates from pigs than from cattle and broilers. The association between the occurrence of resistance and the consumption of the antimicrobial is discussed, as is the occurrence of resistance in other countries. The results of this study show the present level of resistance to antimicrobial agents among a number of bacterial species isolated from food animals in Denmark. Thus, the baseline for comparison with future prospective studies has been established, enabling the determination of trends over time. PMID- 9744763 TI - Different ultrastructural morphology of Pneumocystis carinii derived from mice, rats, and rabbits. AB - Pneumocystis carinii (PC) is a fungus present in the lungs of many mammal species. Even though studies of the genome, the isoenzymes, and the antigens have proved some host-species-linked heterogeneity, the existence of distinct Pneumocystis species or subspecies has still not been accepted. Comparative studies of the ultrastructural morphology of pneumocysts derived from several host species may support evidence of host-species-linked heterogeneity. We have compared the ultrastructural morphology of pneumocysts derived from mice, rats, and rabbits. The density of membrane-limited electron-dense cytoplasmic granules was found to be higher in mouse-derived pneumocysts than in rabbit-derived pneumocysts, and furthermore the average diameter of the granules from mouse pneumocysts was larger than that of granules from rabbit-derived pneumocysts. The average diameter of the filopodia of mouse-derived pneumocysts was smaller than that of filopodia from rat-derived pneumocysts, which was smaller than that of filopodia from rabbit-derived pneumocysts. Globular electron-dense bulbous dilatations at the tip of the filopodia were described for the first time and they were only found on filopodia of mouse-derived pneumocysts. These distinct host-species-linked morphological differences of pneumocysts from mouse, rat, and rabbit may support previous biochemical data indicating the existence of different Pneumocystis species or subspecies. PMID- 9744764 TI - Expression of group II phospholipase A2 in Paneth cells of an adenoma of the rectum. A case report. AB - A case of tubulovillous adenoma in the rectum of a 51-year-old man is presented. The tumour contained numerous Paneth cells which formed well-developed glands in the basal areas. Group II phospholipase A2 and lysozyme were found in the tumour cells by immunohistochemistry. mRNA of group II phospholipase A2 was localized in the tumour cells by in situ hybridization. It was concluded that a considerable part of this rare type of tumour consisted of Paneth cells which were capable of synthesizing group II phospholipase A2. PMID- 9744765 TI - Polychlorinated biphenyl (PCB) exposure produces placental vascular and trophoblastic lesions in the mink (Mustela vison): a light and electron microscopic study. AB - Polychlorinated biphenyls (PCBs) cross the placenta and cause fetal death in mink. No indications of impaired implantation have been reported. To study the effects of PCB on mink placental morphology, 2 groups each of 10 animals were orally exposed to Clophen A50 at 0.65 mg (low dose) and 1.3 mg (high dose) per day for 54 days, starting before mating, with 10 control animals. Placentae from mid to late gestation were examined by light and electron microscopy. In the controls, 11% of placentae were degenerate compared to 31% (low dose) and 64% (high dose) in PCB-exposed mink. All control animals exhibited implantation sites, while one animal in the low dose and four in the high-dose group exhibited none. However, there was no difference between PCB-exposed and control animals in the number of placentation sites in implanted animals. Fetal death was markedly increased in PCB-exposed mink, with only four animals (low dose) having all viable fetuses and eight (low and high dose) having a mixture of viable and dead fetuses. Nine exposed animals displayed maternal vascular lesions in the placental labyrinthine zones of viable fetuses, comprising loss and degeneration of endothelial cells, thrombi and haemorrhages. Extracellular fluid was present between the interstitial layer of maternal vessels and the syncytiotrophoblast, and there was focal degeneration of the trophoblast and fetal vasculature. It appears, therefore, that exposure of the mink placenta to PCBs affects maternal vasculature and produces degenerative changes in the trophoblast and fetal vessels, leading to fetal growth retardation or death. PMID- 9744766 TI - Antibody responses in serum and lung to intranasal immunization with Haemophilus influenzae type b polysaccharide conjugated to cholera toxin B subunit and tetanus toxoid. AB - AIM: Cholera toxin B subunit (CTB) has previously been used as a mucosal carrier for various vaccine candidate antigens. The objective of this study was to see if coupling a bacterial polysaccharide, Haemophilus influenzae type b capsular polysaccharide (HibCPS), to CTB, either directly or through prior coupling to tetanus toxoid (TT), would improve the immunogenicity of HibCPS after nasal immunization. METHODS: HibCPS was conjugated to CTB, TT or via TT to CTB, using glutaraldehyde or 1-ethyl-3(3-dimethylaminopropyl)-carbodiimide (EDAC) and N succinimidyl 3-(2-pyridyldithio) propionate (SPDP). The conjugates were characterized and used for intranasal (IN) and subcutaneous (SC) immunizations of mice. The anti-Hib, -TT and -CTB antibody titers in serum and lungs after the immunizations were measured with ELISA. RESULTS: The HibCTB was poorly immunogenic both given IN and SC compared with HibTT and HibTTCTB, probably because of inefficient coupling. In contrast, the conjugation of CTB to the HibTT conjugate resulted in a preparation which was superior both to the HibTT and the HibCTB conjugates in inducing local IgA and IgG anti-HibCPS antibodies in the lungs. The anti-HibCPS serum IgG titers after IN immunization with the HibTTCTB conjugate were similar to the titers after IN immunization with HibTT, or SC immunization with a commercial HibCRM conjugate vaccine. In contrast to the other conjugates, the HibTTCTB conjugate also gave rise to anti-Hib serum IgA titers. CONCLUSION: We conclude that appropriate conjugation to CTB increases the mucosal immunogenicity of HibCPS, and that intranasal immunization with such a conjugate can give rise to both local and systemic anti-HibCPS antibody responses. PMID- 9744767 TI - Neuroendocrine differentiation and nerves in human adrenal cortex and cortical lesions. AB - Neuroendocrine differentiation and nerve distribution were studied in sections from human cortex (n=11) and cortical lesions (hyperplasias, n=9; adenomas, n=13; carcinomas, n=14) with four markers, namely chromogranin A(CgA), synaptophysin (SYN), neuron-specific enolase (NSE), protein gene product (PGP) 9.5 and small synaptic vesicle protein (SV)2. All but two cases expressed neuroendocrine differentiation. NSE was the most commonly occurring marker and the NSE immunoreactive cells were detected in normal cortex, mainly in zona glomerulosa, as well as in adenomas and carcinomas. SYN and PGP 9.5 immunoreactive cells were especially prominent in the carcinomas, while SV2 immunoreactive cells were seen mainly in normal cortex. The difference in distribution pattern of the neuroendocrine markers between adenomas and carcinomas was not so distinct that it can be used for histopathological diagnosis. The significance of neuroendocrine differentiation in cortex and cortical lesions is uncertain, but may reflect an involvement in special hormonal functions. No obvious relationship was found between the clinical syndromes and the degree of neuroendocrine differentiation. Three of the neuroendocrine markers also visualized nerve structures. PGP 9.5, which is regarded as the most 'general' nerve marker, visualized more nerve structures than did the other markers. Normal cortex contained most immunoreactive nerves, whereas they were less numerous in hyperplasias and sparse or even absent in the neoplasms. The nerves appeared among the parenchymal cells but were particularly prominent around vessels. The results suggest that the cortical nerves influence not only the regulation of the blood supply but also the hormonal regulation at the cellular level. PMID- 9744768 TI - An epitope shared by enterobacterial and neisserial porin proteins. AB - A murine monoclonal antibody (MAb F9-16) raised against a porin protein epitope called Po I of an E. coli 055 strain showed broad cross-reactivity with bacteria within the Enterobacteriaceae, and also recognized neisseriae and moraxellae. In an immunodot assay, the antibody was bound by 32/33 strains of neisseriae and moraxellae after SDS treatment of the bacteria. Testing intact bacteria, 11/33 isolates showed definite MAb binding, including serogroup A and B meningococci. In Western blotting, the anti-Po I MAb targeted the gonococcal porin proteins PIA and PIB, and class 1, class 2, and class 3 porins of meningococci. The MAb showed no reactivity against decapeptides which corresponded to the whole length of a meningococcal class 1 porin protein of the subtype P1, 7, 16. These findings accord with the inference that enterobacterial, neisserial and moraxellae porin proteins share an epitope (Po I) which is determined by the three-dimensional rather than by the primary structure of the proteins and that this epitope is shielded in most isolates but surface-exposed in some isolates, including some strains of meningococci. Since Po I is broadly distributed among commensal and pathogenic bacteria and has demonstrated immunogenicity in humans, this epitope may play a role in elicitation of "normal" antibodies with immunoprotective activity. PMID- 9744769 TI - Determination of flavopiridol (L86 8275; NSC 649890) in human plasma by reversed phase liquid chromatography with electrochemical detection. AB - PURPOSE: Flavopiridol is a flavone which inhibits several cyclin-dependent kinases, and exhibits potent growth-inhibitory activity against a number of human tumor cell lines both in vitro, and when grown as xenografts in mice. It is currently being evaluated in a phase I clinical trial at the National Cancer Institute. The objective of this project was to develop and validate an analytical method for the assay of flavopiridol in human plasma, with sufficient sensitivity to permit the plasma pharmacokinetics of flavopiridol to be studied during clinical trials. METHODS: Flavopiridol was isolated from human plasma samples by extraction with t-butylmethyl ether following alkalinization with borate buffer (pH 8.0). The extract was evaporated, the residue was dissolved in mobile phase, and analyzed by reversed-phase high-pressure liquid chromatography. Chromatography was accomplished with a polymer-based C18 column eluted with a mobile phase consisting of methanol-phosphate buffer, pH 11.0 (53:47 v/v). Electrochemical detection (ECD) was employed. RESULTS: Flavopiridol was recovered from human plasma with an efficiency of 85-87%. Calibration curves were linear over the concentration range 10-500 nM (4.4-219 ng/ml). Plasma standard concentrations were measured with an accuracy and precision ranging from 3.2% to 10%. Regression analysis of flavopiridol concentrations of 15 clinical trial plasma samples ranging in concentration from approximately 50 to 4000 microM quantitated by both ECD and mass spectrometry showed close agreement. The equation of the regression line was y = 1.02x + 8 with a correlation coefficient of 0.969. Continuous infusion of flavopiridol in four patients for 72 h at a rate of 50 mg/m2 per day, resulted in mean steady-state plasma concentrations of from 200 to 300 nM. Levels declined in a biexponential manner following termination of the infusion, falling to approximately 10 nM after 48 h. CONCLUSIONS: An analytical method for the assay of flavopiridol in human plasma was developed with sensitivity to at least 10 nM. The assay is accurate, precise and specific, and is suitable for determination of plasma flavopiridol concentrations for pharmacokinetic studies during clinical trials. PMID- 9744770 TI - Pharmacokinetics and metabolism of thiopurines in children with acute lymphoblastic leukemia receiving 6-thioguanine versus 6-mercaptopurine. AB - Mercaptopurine (6MP) has been the standard drug for maintenance therapy of acute lymphoblastic leukemia. In a multicenter study we investigated whether thioguanine (6TG), which is converted more directly to the cytotoxic thioguanine nucleotides (TGN), offers a therapeutic advantage over 6MP. In this study (COALL 92), 6TG was randomized versus 6MP in maintenance therapy, whereby the doses of both drugs had to be adjusted to a white blood cell (WBC) count of between 2 and 3/nl. In 19 children the plasma levels of both drugs and/or the accumulation of their metabolites in red blood cells (RBC) were measured during intensive treatment in two consecutive chemotherapy blocks, and in 54 children the metabolites in RBC were measured every 3 months during daily treatment in maintenance therapy. There was a marked interindividual difference in the plasma kinetics of the two drugs; after identical doses of 100 mg/m2 an about 4-fold higher peak concentration of the parent drug was reached with 6MP. The main metabolites of 6TG were thioguanine nucleotides (TGN), whereas during 6MP treatment, methylated thioinosine nucleotides (TIN) predominated in erythrocytes. In patients receiving 6TG during maintenance therapy (22 patients) the concentration of methylated TGN reached about 40% of that of unmethylated TGN; after 6MP administration (32 patients) the methylated TIN were concentrated about 26-fold higher in RBC than were TGN. In contrast to 6TG, for 6MP the pattern of metabolites shifted toward the methylated ones with increasing dose. The median TGN concentration was about 7-fold higher in the TG branch, although the median dose was only about 70% of that of 6MP. The WBC values were equivalent in the two treatment groups. Our results suggest that the cytotoxic effect of 6MP is not based solely on the formation of TGN. PMID- 9744771 TI - The benzoquinone ansamycin 17-allylamino-17-demethoxygeldanamycin binds to HSP90 and shares important biologic activities with geldanamycin. AB - PURPOSE: Benzoquinone ansamycins are antibiotics with anticancer potential. First described as tyrosine kinase inhibitors, they are now frequently used to target HSP90 chaperone function. While herbimycin A and geldanamycin (GA) have been widely used in preclinical studies, both drugs are poor candidates for clinical trials owing to their in vivo toxicity and lack of stability. We therefore examined the biologic effects of 17-allylamino-17-demethoxygeldanamycin (17-AG), an ansamycin derivative with lower in vivo toxicity than GA. METHODS: Binding of 17-AG to HSP90 was studied in vitro using a GA-affinity beads competition assay. We analyzed the drug-induced destabilization of p185erbB2, Raf-1 and mutant p53 in SKBR3 breast cancer cells by Western blotting. The antiproliferative activities of 17-AG and GA were compared using the MTT assay. RESULTS: We found that, in a similar manner to GA itself, 17-AG bound specifically to HSP90. It also led to degradation of the receptor tyrosine kinase p185erbB2, the serine/threonine kinase Raf-1 and mutant p53. Both GA and 17-AG displayed comparable antiproliferative effects in SKBR3 and MCF7 cells. Even though HSP90 binding by 17-AG was weaker than by GA, 17-AG and GA caused biologic effects in tumor cells at similar doses. CONCLUSION: 17-AG shares the important biologic features of its parent compound GA. Since 17-AG has a better toxicity profile than GA, it is an interesting candidate benzoquinone ansamycin for clinical development. PMID- 9744772 TI - Inhibition of intestinal microflora beta-glucuronidase modifies the distribution of the active metabolite of the antitumor agent, irinotecan hydrochloride (CPT 11) in rats. AB - PURPOSE: SN-38, a metabolite of irinotecan hydrochloride (CPT-11), is considered to play a key role in the development of diarrhea as well as in the antitumor activity of CPT-11. We have previously found that the inhibition of beta glucuronidase, which hydrolyzes detoxified SN-38 (SN-38 glucuronide) to reform SN 38, in the lumen by eliminating the intestinal microflora with antibiotics, markedly ameliorates the intestinal toxicity of CPT-11 in rats. In this study we compared the disposition of CPT-11 and its metabolites in rats treated with and without antibiotics. METHODS: Rats were given drinking water containing 1 mg/ml penicillin and 2 mg/ml streptomycin from 5 days before the administration of CPT 11 (60 mg/kg i.v.) and throughout the experiment. CPT-11, SN-38 glucuronide and SN-38 concentrations in the blood, intestinal tissues and intestinal luminal contents were determined by HPLC. RESULTS: Antibiotics had little or no effect on the pharmacokinetics of CPT-11, SN-38 glucuronide or SN-38 in the blood, or in the tissues or contents of the small intestine, which has less beta-glucuronidase activity in its luminal contents. In contrast, antibiotics markedly reduced the AUC1-24 h of SN-38 (by about 85%) in the large intestine tissue without changing that of CPT-11, and this was accompanied by a complete inhibition of the deconjugation of SN-38 glucuronide in the luminal contents. CONCLUSIONS: These results suggest that SN-38, which results from the hydrolysis of SN-38 glucuronide by beta-glucuronidase in the intestinal microflora, contributes considerably to the distribution of SN-38 in the large intestine tissue, and that inhibition of the beta-glucuronidase activity by antibiotics results in decreased accumulation of SN-38 in the large intestine. PMID- 9744773 TI - Prevention of apomorphine- or cisplatin-induced emesis in the dog by a combination of methylnaltrexone and morphine. AB - PURPOSE: Morphine can have either an emetic or an antiemetic effect. The emetic effect of morphine can be blocked by methylnaltrexone (MNTX), a quaternary opioid antagonist with peripheral action. In this study, we tested the hypothesis that administering MNTX to block the peripheral emetic effect of morphine would unmask the central antiemetic effect of the morphine. The net result, we hypothesized, would be a reduction in apomorphine- or cisplatin-induced emesis. METHODS: MNTX 0.25 mg/kg and morphine 1 mg/kg were administered to conscious dogs which were then challenged with the potent emetic agents apomorphine or cisplatin. Emesis was assessed by the presence of characteristic retching motions accompanied by the regurgitation of gastric contents. RESULTS: We observed that apomorphine challenges of 0.1 mg/kg and of 0.03 mg/kg produced 100% emesis in control animals. After pretreatment with MNTX and morphine, the frequency of emesis with the larger dose of apomorphine was reduced to 50% and with the smaller dose to 22%. MNTX alone did not block apomorphine-induced emesis. In animals challenged with cisplatin 3 mg/kg, the emetic response was 100%. Emesis did not occur in animals pretreated with MNTX 0.25 mg/kg and morphine 1 mg/kg before cisplatin. CONCLUSIONS: Our results demonstrate that MNTX combined with morphine reduces apomorphine-induced emesis and blocks cisplatin-induced emesis. These results support the hypothesis that the emetic effect of morphine is peripheral and its antiemetic action is central. In combination, MNTX and morphine may have a clinical role in the treatment of chemotherapy-induced emesis. PMID- 9744774 TI - Fluorescence methods to assess multidrug resistance in individual cells. AB - PURPOSE: Microscopic methods to measure the activity of drug extrusion systems important in multidrug resistance in individual cells were developed. METHODS: Multidrug-resistant (MDR) and parental lines of hamster CHO and pituitary GH3 cells were incubated with the acetoxymethylester (AM) forms of several fluorescent calcium-sensing dyes, fura2, indo1 and fluo3. The AM forms of these compounds are hydrolyzed by intracellular esterases and then trapped in cells, and the AM forms of the dyes are excellent substrates for P-glycoprotein (Pgp). RESULTS: The fluorescent free acid forms of fura2, indol and fluo3 did not accumulate in MDR lines unless a chemosensitizer such as cyclosporin A, R(+)verapamil, quinidine, or progesterone was included during loading to prevent the cells from extruding the AM forms of the dyes before they could be hydrolyzed. Cyclosporin A increased the fluorescence due to intracellularly trapped fura2 free acid from 8- to 20-fold and was maximally effective at < 5 microM. Fluorescence microscopy was employed to measure fura2 free acid accumulation by parental and MDR cell lines using excitation at the Ca2+ insensitive wavelength. When MDR cells were incubated with rhodamine 123 and fura2/AM, no fluorescence was detectable. Cellular fluorescence was dramatically increased by inclusion of cyclosporin A, quinidine, progesterone, or R(+)verapamil. There was no measurable decline in the fura2 free acid fluorescence in 1 h while the fluorescence due to rhodamine 123 diminished rapidly in cells overexpressing Pgp. CONCLUSIONS: These fluorescence methods detect drug-extruding activity in individual cells and therefore have the potential to provide complementary information to studies quantifying protein or mRNA levels of Pgp or other efflux pumps. In addition, they provide a rapid and quantifiable method for screening multidrug resistance reversing agents. PMID- 9744775 TI - Pharmacokinetics, antifolate activity and tissue distribution of PT523 in SCC VII tumor-bearing mice. AB - PURPOSE: To monitor the pharmacokinetics of PT523 and methotrexate in C3H mice with transplanted SCC VII tumors; to compare the impact of PT523 and methotrexate on tumor and normal host 5,10-methylenetetrahydrofolate levels; and to synthesize [14C]PT523 and determine its time-dependent tissue distribution in tumor and host tissues. METHODS: C3H mice bearing SCC VII tumors were given i.p. PT523 or methotrexate. Plasma drug levels and tumor, gut and marrow 5,10 methylenetetrahydrofolate were assayed. [14C]PT523 was synthesized and administered i.v. to tumor-bearing mice for tissue distribution analysis. RESULTS: Areas under the curve, mean residence times, whole body clearances, apparent distribution volumes, and plasma protein binding of PT523 vs methotrexate were, respectively, 4311 vs 6472 microM x min(-1); 20 vs 16 min; 0.56 vs 0.36 ml min(-1); 532 vs 325 ml x kg(-1); and 70% vs 30%. Both PT523 and methotrexate caused time-dependent declines in 5,10-methylenetetrahydrofolate in tumor and marrow, but not in gut mucosa [corrected]. Gut levels began to recover within 4 h in the PT523-treated group only. [14C]PT523 distributed mainly into the liver, duodenum, kidneys, lungs, tumor, pancreas and muscle; less into the spleen, blood cells, heart, brain and testicles; and very little into gut [corrected. Only 35% of the dose was excreted, and 2.9-fold more in feces than urine. CONCLUSIONS: Despite its more rapid clearance, accumulation of PT523 in extravascular tissues was greater than that of methotrexate. Consequently, less PT523 was recovered in feces and urine and its apparent volume of distribution was greater. PT523 selectively depleted 5,10-methylenetetrahydrofolate pools in tumor and, less persistently, in marrow, but spared the gut mucosa [corrected]. [14C]PT523 tissue distribution correlated with organ mass and blood supply. PMID- 9744776 TI - Prediction of carboplatin clearance calculated by patient characteristics or 24 hour creatinine clearance: a comparison of the performance of three formulae. AB - PURPOSE: Carboplatin doses can be individualized using the formula of Calvert et al. (Calvert formula) dose (mg) = area under the plasma concentration versus time curve (AUC) x [glomerular filtration rate (GFR) + 25]. Creatinine clearance (Ccr), either measured by the 24-h method or calculated by the formula of Cockcroft and Gault [Cockcroft-Gault (CG) formula], is often substituted for the GFR. The CG formula is based on patient weight, age and sex, and the serum creatinine (Cr) concentration. Another method for predicting carboplatin clearance (CL) using patient characteristics has also been proposed by Chatelut et al. (Chatelut formula). This study was undertaken to evaluate the performance of the three formulae in predicting standard- and low-dose carboplatin pharmacokinetics. METHODS: A total of 52 patients with advanced lung cancer were enrolled in this pharmacokinetic study; 37 received standard-dose carboplatin and 25 received low-dose carboplatin. The Cr concentration was measured using an enzymatic assay. The three formulae were used to predict carboplatin CL. The median absolute percent error (MAPE) for each formula was evaluated by comparing the calculated and observed CL. For comparison of AUCs, free platinum plasma concentrations were measured at intervals up to 24 h after carboplatin administration. AUCs were determined and compared with predicted values. RESULTS: In the standard-dose carboplatin group, the MAPEs for the prediction of carboplatin CL from the 24-h Calvert, CG-Calvert and Chatelut formulae were 13%, 12% and 23%, respectively. In the low-dose carboplatin group, the corresponding MAPEs were 27%, 18% and 44%, respectively. Observed standard-dose carboplatin AUCs after aiming for target AUCs of 5 and 6 mg x min/ml using the Calvert formula based upon the 24-h Ccr were 5.3+/-0.8 and 5.9+/-0.8, respectively, indicating a small and acceptable bias compared with that predicted from the dosing formula. CONCLUSIONS: The pharmacokinetics of standard-dose carboplatin were accurately predicted by the Calvert formula based upon either 24-h or CG calculated Ccr, but not by the Chatelut formula. Either CG-calculated or 24-h Ccr can be substituted for the GFR in the Calvert formula for the determination of individual doses. The poor predictability of the Chatelut formula found in this study might be the result of a differences in either the Cr assay or the patient population. Therefore, formulae which attempt to estimate GFR are not necessarily valid if either the Cr assay or the patient population is changed. PMID- 9744777 TI - A new analogue of 10-deazaaminopterin with markedly enhanced curative effects against human tumor xenografts in mice. AB - PURPOSE: These studies sought to evaluate the biochemical and cellular pharmacokinetic properties, cytotoxicity and antitumor efficacy of a new analogue of 10-deaza-aminopterin (PDX) against human tumors. METHODS: Studies were conducted with a group of human tumor cell lines in culture examining PDX and other folate analogues as permeants for mediated membrane transport, as inhibitors of dihdrofolate reductase and as substrates for folylpolyglutamate synthetase. These same analogues were examined for their cytotoxicity following a 3-h pulse exposure, in experiments providing a value for IC50. Other studies with these analogues were conducted in nude mice bearing subcutaneously implanted human tumors. Treatment of the mice was initiated 4 days after implantation of the tumor using a schedule of administration of one dose per day for 5 days. The tumors were measured 6 days after cessation of therapy and compared to controls for assessment of response. RESULTS: In the CCRF-CEM cell system, PDX was 2- to 3 fold less effective as an inhibitor of dihydrofolate reductase than aminopterin (AMT), methotrexate (MTX) or edatrexate (EDX) but much more effective as a permeant for one-carbon, reduced folate transport inward (PDX > AMT approximately equal to EDX > MTX) and substrate for folylpolyglutamate synthetase (PDX > AMT > EDX > MTX). As predicted by these results, PDX was 15- to 40-fold more cytotoxic than MTX and 3- to 4-fold more cytotoxic than the highly potent EDX following a 3 h pulse exposure in culture of CCRF-CEM cells and cells from a panel of three human breast and two human nonsmall-cell (NSC) lung cancers. The same relative differences were shown for the therapeutic efficacy of these three analogues at equitoxic doses in studies with the human MX-1 and LX-1 tumors and the human A549 NSC lung tumor xenografted in nude mice. On a schedule of qd x 5 given 3-4 days posttransplant, MTX was minimally active (modest tumor growth delay) against all three tumors. EDX was highly active (25-35% complete regressions and 5-10% cures) against the MX-1 and LX-1 tumors but very modestly active (no regressions) against the A549 tumor. In contrast, PDX was even more active (75-85% complete regressions and 25-30% cures) than EDX against the MX-1 and LX-1 tumors and highly active (30% complete regressions and 20% cures) against the A549 tumor. CONCLUSIONS: These studies showed significantly enhanced antitumor properties of PDX compared with MTX and EDX. Based upon these results, clinical trials of PDX in patients with metastatic breast and NSC lung cancer appear to be warranted. PMID- 9744778 TI - Quaternary ammonium analogs of ether lipids inhibit the activation of protein kinase C and the growth of human leukemia cell lines. AB - PURPOSE: ET-18-OCH3 (1-O-octadecyl-2-O-methyl-rac-glycero-3-phosphocholine) is a representative of the first generation of antitumor ether lipids and is a model in the development of new compounds including a series of quaternary ammonium analogs (QAA). In the present study, we evaluated the QAA as inhibitors of cell growth and studied their mechanism of action. METHODS: We compared the effects of the QAA on the proliferation of human leukemia cell lines which are sensitive (HL 60) or resistant to ET-18-OCH3 (HL-60R and K562). In addition we used cell fractionation and enzymatic assays to determine the effects of QAA on protein kinase C (PKC) translocation in response to 12-O-tetradecanoyl-phorbol-13-acetate (TPA). RESULTS: The QAA and ET-18-OCH3 were approximately equally effective inhibitors of HL-60 cell growth. However, the QAA were more effective inhibitors of K562 and HL-60R cell proliferation. The HL-60R cells, which were selected for resistance to ET-18-OCH3, were also resistant to BM 41.440 which is structurally similar. In serum-free medium, the phosphorus-containing compounds (ET-18-OCH3, BM 41.440 and He-PC) were much more effective inhibitors (8-20-fold) of the K562 cell line while the activities of the QAA were only moderately increased (1.2-2.3 fold). When serum albumin was added to the serum-free medium, the K562 cells became resistant to ET-18-OCH3, suggesting that albumin is responsible for the differential sensitivity. The QAA compounds, which inhibit PKC activity in vitro, inhibited cell proliferation. However, a QAA which did not inhibit PKC did not inhibit cell proliferation. The phorbol ester TPA stimulates PKC translocation and causes HL-60 cell differentiation to adherent macrophage-type cells. The QAA inhibited TPA-induced cell differentiation and PKC translocation indicating that they also inhibit PKC in intact cells. CONCLUSIONS: The cellular effects of the QAA appear to be due to inhibition of PKC. In addition, these data indicate that albumin, which is important as a mediator of the uptake of ET-18-OCH3, has only a small effect on the uptake of QAA. Together these data indicate that the QAA are potential anticancer agents, showing a significant ability to inhibit growth of leukemia cell lines that are resistant to ET-18-OCH3. PMID- 9744779 TI - Sequence effect of irinotecan (CPT-11) and topoisomerase II inhibitors in vivo. AB - The DNA topoisomerases I and II are the target of several clinically important antineoplastic agents which produce DNA cleavage by stabilization of the covalent DNA-protein bond with resultant cell death after DNA synthesis is attempted. Depletion of the target topoisomerase and reciprocal changes in the other occur with drug treatment. PURPOSE AND METHODS: To develop empiric treatment regimens of combinations and sequences of agents directed against topoisomerase I (irinotecan/CPT-11) and II (etoposide and doxorubicin), in vivo studies were performed in mice bearing the EMT-6 mammary tumor to assess efficacy, host tolerance and the resultant biochemical changes in topoisomerase mRNA and protein. RESULTS: At 24 h after therapy, depletion of the target topoisomerase mRNA and protein with reciprocal increases in the alternate topoisomerase mRNA and, to a lesser extent, protein were noted. No therapeutic antagonism was found with any combination or sequence of agents, and therapeutic antagonism was noted with concurrent irinotecan/etoposide and sequential doxorubicin/irinotecan. Depletion of target topoisomerases by combined therapy beyond a threshold necessary for therapeutic efficacy produced no additional benefit. CONCLUSIONS: Antineoplastic therapy with combinations of topoisomerase I and II agents is feasible and may produce therapeutic synergy. The appropriate sequence may depend on the particular agents used. The rationale for such therapy, that topoisomerases I and II may have reciprocal and compensatory interactions, is supported by the biochemical data. PMID- 9744780 TI - Advanced colorectal cancer in the elderly: results of consecutive trials with 5 fluorouracil-based chemotherapy. AB - To evaluate toxicity and efficacy of chemotherapy in elderly patients (> or = 65 years of age) with advanced colorectal cancer, data from two consecutive trials conducted between 1984 and 1995 at the National Institute for Cancer Research were analysed comparing the results of treatment in those 65 years of age or older and in those younger than 65 years. Of 215 patients recruited, 82 elderly patients (median age 70 years, median performance status 1) received one of the following regimens based on 5-fluorouracil (5-FU): (1) weekly 5-FU 600 mg/m2 i.v. bolus (30 patients); (2) weekly 5-FU 600 mg/m bolus plus leucovorin (LV) 500 mg/m2 2-h i.v. infusion (28 patients); (3) Weekly 5-FU 2600 mg/m2 24-h continuous i.v. infusion plus LV 100 mg 4-h i.v. infusion and 50 mg orally every 4 h for five doses (24 patients). Overall, 1071 chemotherapy cycles were administered with a median number of 12 courses per patient. The main side effects were diarrhoea, observed in 38% of patients, stomatitis in 24% of patients and hand foot syndrome in 13% of patients, and haematological toxicity affected only 15% of patients. No patient suffered grade IV toxicity. In three patients chemotherapy was discontinued because of toxicity (two patients suffered grade III diarrhoea, one patient grade III hand-foot syndrome). No significant difference in toxicity was evident between patients older than or younger than 65 years. Analysis of median dose intensity demonstrated no difference between the two groups. Overall objective response was observed in 18% (95% confidence limits 11-29) of elderly patients (15/82) in comparison with 23% (95% CL 17-32) of patients < 65 years of age (31/133 pts). In conclusion, chemotherapy in elderly patients with advanced colorectal cancer is a safe and effective treatment with acceptable toxicity and comparable objective response rates. PMID- 9744781 TI - Superiority of hepatic arterial infusion in preventing catabolism of 5-FU compared with portal vein infusion revealed by an in vivo 19F NMR study. AB - PURPOSE: The aim of this study was to identify the route of administration of 5 FU with the greatest pharmacological advantage in a rat model using non-invasive in vivo 19F nuclear magnetic resonance (NMR) spectroscopy. METHODS: 5-FU (50 mg/kg) was administered to anesthetized Wistar rats cannulated into the hepatic artery, portal vein or tail vein and 11 NMR spectra were acquired from the liver region to 60.5 min every 5.5 min. RESULTS: With systemic i.v. (tail vein) infusion, the 19F-NMR signal for 5-FU from the liver region peaked in the first spectrum (0-5.5 min), and then gradually decreased. The signal for the 5-FU catabolite alpha-fluoro-beta-alanine (FBAL) gradually increased to the sixth spectrum (0-33.0 min) and then plateaued. Following portal vein infusion the intensity of the first 5-FU spectrum was twice as high as that following i.v. infusion, but the intensity decreased and the FBAL signal increased gradually in the sixth spectrum as systemic i.v. infusion. In contrast, the intensity of the 5 FU signal following hepatic artery infusion was the same as that following portal vein infusion in the first spectrum, and maintained a strong intensity to the final spectrum (60.5 min). The FBAL signal was detected from the second spectrum following hepatic artery infusion, but its intensity was significantly weaker than that following i.v. or portal vein infusion. CONCLUSIONS: Hepatic arterial infusion resulted in the active form of 5-FU being present for a longer time and its degradation in the liver being suppressed compared with the results following portal vein infusion. This catabolic advantage of hepatic arterial infusion could lead to a more potent anti-tumor activity against liver metastases, but could also lead to significant host toxicity including biliary toxicity. We recommend that the dose/schedule of 5-FU administered via the hepatic artery should be adjusted carefully. PMID- 9744782 TI - Effect of electrical stimulation on stress and urge urinary incontinence. Clinical outcome and practical recommendations based on randomized controlled trials. AB - BACKGROUND: The aim of the present study was to review the literature on randomized controlled trials of electrical stimulation to treat urge and stress urinary incontinence. METHODS: Studies were compiled from Medline from 1980 till 1996 and manual searches of relevant journals. Randomized controlled studies full length published in English, German and Scandinavian languages were included. RESULTS: Nine studies evaluating the effect of electrical stimulation on stress urinary incontinence and one study evaluating the effect of urge incontinence were found. Only three studies had a sufficient sample size to enable conclusion on stress urinary incontinence. Two demonstrated negative, and one positive effect (20%) cure and 46% improved measured by pad test). The study on urge incontinence demonstrated that there was no difference in effect after electrical stimulation or anticholinergic drugs. CONCLUSION: The results of randomized controlled trials evaluating the effect of electrical stimulation to treat stress and urge urinary incontinence are conflicting. There is a need for more randomized controlled trials with sufficient sample sizes, use of sensitive, reproducible and valid outcome measures, and optimal stimulation parameters. Based on the present knowledge pelvic floor muscle exercise should be the first choice of treatment for stress urinary incontinence. PMID- 9744783 TI - Evaluation of the subjective and objective effect of maximal electrical stimulation in patients complaining of urge incontinence. AB - BACKGROUND: Favorable results have been reported following Maximal Electrical Stimulation (MES) of patients with urgency and urge incontinence. However, patient groups have often been mixed and outcome measures poorly defined. We therefore wanted to treat a homogeneous patient population with MES and evaluate the effect by defined subjective and objective outcome measures. METHODS: Eighteen female patients complaining of urge incontinence had MES performed. Before and 3 months after MES, the patients performed a 24 hour micturition chart and pad test. They indicated on a visual analogue scale their subjective degree of urgency and leakage and had an ambulatory urodynamic monitoring performed. Nine months after MES the patients were asked whether their urge incontinence was less, equal or more troublesome than before MES. RESULTS: After MES the patients indicated significantly less urgency and leakage. A significant difference was not found in any of the objective outcome measures after MES. Six out of 18 patients (33%) found their urge incontinence less troublesome 9 months after MES, while 12 (66%) found it unchanged or more troublesome than before. CONCLUSIONS: Significantly subjective effect was found following MES. However, none of the objective outcome measures were significantly improved. We were disappointed by the results and have stopped using the method. PMID- 9744785 TI - Long-term effects ten years after maximal electrostimulation of the pelvic floor in women with unstable detrusor and urge incontinence. AB - OBJECTIVE: The purpose was to study any long-term therapeutic effects of maximal electrical stimulation in female urge incontinence. METHODS: A postal questionnaire containing six questions about urinary incontinence was distributed to 30 women who had been treated with maximal stimulation because of unstable detrusor and urge incontinence 9-13 years earlier. The response rate was 90% (27 women). The mean age at follow-up was 62 years. RESULTS: Twenty-one (78%) women reported symptoms of urge incontinence. Among them, 13 had this problem daily, whereas eight only had problems weekly or even more seldom. Nineteen (70%) women reported symptoms of stress incontinence. Twenty-one women would have recommended maximal stimulation to a friend today. CONCLUSION: After approximately ten years most of the women had symptoms of urge incontinence. This was, however, a minor problem among a third of them. A majority of the women were satisfied with maximal stimulation as a treatment modality. The treatment had not prevented a later occurrence of stress incontinence. PMID- 9744786 TI - Aspects on the actual practice of surgical management of urinary incontinence in Norway and Finland. AB - OBJECTIVE: To elucidate aspects of the actual practice of surgical management of urinary incontinence in Norway and Finland. METHOD: A questionnaire about preoperative diagnostic procedures, type of surgery, follow-up procedure and number of operations per center and per surgeon per year (1995) was sent to all departments of Gynecology and Obstetrics in Norway and Finland. RESULT: Thirty one (86%) of the Norwegian and 26 (90%) of the Finnish departments returned the questionnaire. More than 90% of the departments performed preoperative urodynamic evaluation. Burch colposuspension was used by more than 90% of the departments in the two countries. Generally the number of procedures per center and per department was low. Twenty-nine and 12% of the departments respectively did not perform any follow-up after surgery. Only 16% of the Norwegian and 23% of the Finnish centers had controlled results of surgery. CONCLUSION: There seems to be a need for criteria of good clinical practice or a reference program for all steps of surgical management of stress incontinence in order to stimulate the process of quality development of this treatment. PMID- 9744784 TI - Advantages and pitfalls of functional electrical stimulation. AB - Functional electrical stimulation has many theoretical advantages. In clinical practice, very favorable results have been repeatedly presented. The experience now encompasses thirty years and a very large number of incontinent patients. Although the methods are widely used, they are differently appreciated. Problems include the fact that functional electrical stimulation does not belong to the therapeutic traditions in urology and gynecology, there is a need of personal training for successful treatment and there is a lack of systematic studies on different clinical applications. Significant advantages are a rational physiological basis, applicability in a variety of lower urinary tract dysfunctions, few side effects and a potential curative effect. PMID- 9744787 TI - Laparoscopic Burch colposuspension. AB - OBJECTIVE: To review the available information on laparoscopic colposuspension. METHOD AND MATERIAL: One randomized controlled trial, four non-randomized comparative studies and ten open observational clinical series were reviewed. RESULTS: The available information was generally poor characterized by the small size of series, considerable variation in surgical techniques, short follow-up and mainly subjective assessment of outcome. CONCLUSIONS: Laparoscopic colposuspension has been shown to be technically feasible but the therapeutic usability of the procedure remains to be documented. There is an urgent need for some large, rigorous prospective randomized studies PMID- 9744788 TI - The tensionfree vaginal tape procedure (TVT) for treatment of female urinary incontinence. A minimal invasive surgical procedure. AB - BACKGROUND: To test the suitability of a new surgical procedure for treatment of female urinary incontinence to be used as an ambulatory and minimal invasive operation. METHODS: Thirty-one consecutive patients with urodynamically proven stress incontinence had a tensionfree vaginal tape procedure performed. Operation time, the amount of anesthetics and analgetics used, postoperative mobilization, voiding patterns, residual urine volumes, per- and postoperative complications, hospital stay and need for sick leave were prospectively recorded. RESULTS: All 31 patients were cured from stress incontinence. Local anesthesia was used in all cases and additional analgetics were needed in only small doses. Seventy per cent of the patients were released from the hospital on the same day of the operation. By medical criteria 90% could have been released on the same day. No significant per- or postoperative complications occurred. Three patients needed postoperative catheterization. All but one patient was able to empty her bladder within 24 hours from the operation. An average of 15 days sick leave was prescribed. CONCLUSION: The tensionfree vaginal tape procedure seems to fulfil the criteria for being regarded as a minimal invasive surgical procedure for treatment of female urinary stress incontinence. It is highly effective and is associated with very few intra and postoperative side effects. PMID- 9744789 TI - Transvaginal needle bladder neck suspension for stress urinary incontinence: practicable methods but not optimal results. AB - BACKGROUND: To describe the principal methods of needle bladder neck suspension including complications and to evaluate their cure rates. METHODS: The methods are described according to the original papers of Pereyra, Stamey and Raz. Figures of complications and cure rates are based on recent reviews and prospective studies. RESULTS: The complication rates do not exceed those of other surgical procedures for stress incontinence. The frequencies of pain and suture removal, voiding disorder, and de novo detrusor instability are each in the range of 5-6%. Objectively assessed cure rates of the endoscopic needle bladder neck suspension are 87% after one year and for the non-endoscopic methods 70%. However, prospective studies have shown that this cure rates may deteriorate to approximately 40% after five years compared to 82% after the Burch colposupension. CONCLUSION: The needle procedures should be reserved to patients who can only tolerate a minor surgical procedure and accept the risk of failure after a few years. PMID- 9744790 TI - Transurethral endoscopic treatment of urinary stress incontinence in women. Materials and results in former and present agents. PMID- 9744791 TI - Cse1p functions as the nuclear export receptor for importin alpha in yeast. AB - CSE1 is essential for yeast cell viability and has been implicated in chromosome segregation. Based on its sequence similarity, Cse1p has been grouped into the family of importin beta-like nucleocytoplasmic transport receptors with highest homology to the recently identified human nuclear export receptor for importin alpha, CAS. We demonstrate here that Cse1p physically interacts with yeast Ran and yeast importin alpha (Srp1p) in the yeast two-hybrid system and that recombinant Cse1p, Srp1p and Ran-GTP form a trimeric complex in vitro. Re-export of Srp1p from the nucleus into the cytoplasm and nuclear uptake of a reporter protein containing a classical NLS are inhibited in a cse1 mutant strain. These findings suggest that Cse1p is the exportin of importin alpha in yeast. PMID- 9744792 TI - Partial purification of a GTP-insensitive (1-->3)-beta-glucan synthase from Phytophthora sojae. AB - A (1 --> 3)-beta-glucan synthase activity was identified in cell membrane preparations from the oomycete Phytophthora sojae, a soybean pathogen. The activity could be solubilized using the zwitterionic detergent CHAPS at relatively low concentrations (3 mg/ml). High salt concentrations were not effective in removing the activity from the membranes. Detergent solubilization of the enzyme resulted in a six-fold increase of calculated Vmax values (2.5 vs. 0.4 nkat/mg protein) but only minor alteration of the Km (10.6 vs. 10.7 mM). Analysis of the reaction product of the solubilized enzyme by enzymatic degradation and by 2D NMR spectroscopy confirmed its identity as a linear high molecular weight (1 --> 3)-beta-glucan. Glucan synthase activity in both membrane and solubilized preparations was not activated by GTP or divalent cations as reported for other fungal or plant glucan synthases, The activity was inhibited, as expected, in a competitive manner by UDP with a Ki of 2.9 mM. Partial purification of the enzyme was achieved by anion exchange chromatography followed by product entrapment. This procedure resulted in the selective enrichment of a protein band with apparent Mr 108,000 in SDS-PAGE which was not visible in any of the steps preceding product entrapment. The glucan pellets from product entrapment contained up to 3% of the initial enzyme activity present in the fraction used for the procedure. PMID- 9744793 TI - Enzymatic repair of oxidative damage to human apolipoprotein A-I. AB - Oxidative damage to apolipoprotein A-I that occurs in vivo commonly involves methionine oxidation, and is accompanied by alterations in structure, lipid association, and cholesterol efflux function. We have used the enzyme peptide methionine sulfoxide reductase (PMSR) to reverse this damage, and shown by a variety of criteria that enzyme treatment restores the primary, secondary, and tertiary structure and lipid association characteristic of the native unoxidized protein. Lipid-associated as well as lipid-free apolipoprotein A-I reacts with PMSR, suggesting that enzymatic reduction of oxidized apolipoprotein A-I in high density lipoproteins can result in restoration of biological activity and the ability to promote cholesterol efflux from cells. PMID- 9744794 TI - Phosphorylation of specific sets of tau isoforms reflects different neurofibrillary degeneration processes. AB - Tau proteins are the basic components of filaments that accumulate within neurons during neurofibrillary degeneration, a degenerating process with disease-specific phenotypes. This specificity is likely to be sustained by both phosphorylation state and isoform content of tau aggregates that form neuronal inclusions. In the present study, characterization of tau isoforms involved in neurofibrillary degeneration in Alzheimer's disease, Pick's disease, corticobasal degeneration and progressive supranuclear palsy was performed. Both analyses by immunoblotting using specific tau antibodies and cell transfection by tau isoform cDNAs allowed us to demonstrate the aggregation of (1) the six hyperphosphorylated tau isoforms in Alzheimer's disease, (2) tau isoforms without exon 10-encoding sequence in Pick's disease and (3) hyperphosphorylated exon 10-tau isoforms in corticobasal degeneration and progressive supranuclear palsy. Thus, neurofibrillary degeneration phenotypes are likely to be related to the phosphorylation of different combinations of tau isoforms (with and/or without exon 10-encoding sequence) in subpopulations of neurons. PMID- 9744795 TI - Drosophila melanogaster acylphosphatase: a common ancestor for acylphosphatase isoenzymes of vertebrate species. AB - An open reading frame encoding a putative acylphosphatase was found in Drosophila melanogaster. The corresponding gene product shows 40% identity and 22 additional amino acid residues at the C-terminus as compared to muscle- and common-type human acylphosphatases. Moreover, all the residues involved in the catalytic mechanism of vertebrate enzymes are conserved in the D. melanogaster acylphosphatase. The D. melanogaster protein and a deletion mutant, similar in length to vertebrate acylphosphatases, were produced by cloning the corresponding cDNA in Escherichia coli. The wild-type enzyme is a protein with a well established three-dimensional fold and a markedly reduced conformational stability as compared to vertebrate isoenzymes. The specific activity of the enzyme is significantly lower than that found in vertebrate enzymes though the substrate binding capability is basically unaltered. The deletion of 22 residues does not cause a significant change in k(cat), while affecting the apparent binding parameters. This work suggests that the genes encoding the vertebrate enzymes originate from an ancestor gene by duplication and subsequent evolution. PMID- 9744796 TI - A role of Lys614 in the sulfotransferase activity of human heparan sulfate N deacetylase/N-sulfotransferase. AB - An active sulfotransferase (ST, residues 558-882) domain of the human heparan sulfate N-deacetylase/N-sulfotransferase (hHSNST) has been identified by aligning the amino acid sequence of hHSNST to that of mouse estrogen sulfotransferase (EST). The bacterially expressed ST domain transfers the 5'-sulfuryl group of 3' phosphoadenosine-5'-phosphosulfate (PAPS) to only deacetylated heparin with an efficiency similar to that previously reported for the purified rat HSNST. Moreover, the K(m,PAPS) (2.1 microM) of the ST domain is also similar to that of the rat enzyme. Lys48 is a key residue in mEST catalysis. The residue corresponding to Lys48 is conserved in all known heparan sulfate sulfotransferases (Lys614 in the ST domain of hHSNST). Mutation of Lys614 to Ala abolishes N-sulfotransferase activity, indicating an important catalytic role of Lys614 in the ST domain. Crystals of the ST domain have been grown (orthorhombic space group P2(1)2(1)2) with diffraction to 2.5 A resolution. PMID- 9744797 TI - Macrophage populations of different origins have distinct susceptibilities to lipid peroxidation induced by beta-haematin (malaria pigment). AB - We investigated the susceptibility of peritoneal mouse macrophages and macrophage and microglia cell lines to the peroxidative activity of beta-haematin, the synthetic polymer identical to native malaria pigment. The extent of lipid peroxidation, measured as production of thiobarbituric acid reactive substances (TBARS), was greater for peritoneal macrophages than for cell lines and microglia cells. TBARS production apparently was not attributable to the release of free iron from the protoporphyrin moiety, but related to lower glutathione content and different lipid composition of the cell membrane. These findings offer a new interpretation for the contentious immunomodulatory effects of beta-haematin reported for phagocytes of different origins. PMID- 9744798 TI - Purification and characterization of leukotriene A4 hydrolase from Xenopus laevis oocytes. AB - In mammals, leukotriene A4 hydrolase converts leukotriene A4 into the proinflammatory mediator leukotriene B4. We have purified and characterized a non mammalian leukotriene A4 hydrolase from Xenopus laevis oocytes. This enzyme contains one zinc atom and catalyzes an anion-dependent peptidase activity, two key features of the mammalian enzymes. The amino acid sequence of an internal segment is 60% identical with human leukotriene A4 hydrolase but only 27% identical with rat aminopeptidase B. The Xenopus laevis enzyme is catalytically very efficient and, unlike the human enzyme, converts leukotriene A4 into two enzymatic metabolites, viz. leukotriene B4 and delta6-trans-delta8-cis leukotriene B4. PMID- 9744799 TI - Functional reassembly of the anion transport domain of human red cell band 3 (AE1) from multiple and non-complementary fragments. AB - We constructed cDNA clones encoding N-terminal, C-terminal and internal polypeptide fragments of the human red cell anion exchanger (band 3; AE1). The internal fragments comprised between one and seven putative transmembrane spans with two or more spans deleted from both termini of the membrane domain of band 3. Sets of three, four or five complementary fragments, which together represented the complete amino acid sequence of the membrane domain, were co expressed in Xenopus oocytes. Stilbene disulphonate-sensitive chloride uptake assays revealed that all six of the three-fragment combinations and two of the four-fragment combinations reassembled functionally in vivo. Unexpectedly, co expression of a non-complementary pair of fragments comprising the first five and last seven putative transmembrane spans (i.e. entirely lacking spans six and seven) was also found to be sufficient to generate stilbene disulphonate sensitive chloride uptake. PMID- 9744800 TI - Caenorhabditis elegans small heat-shock proteins Hsp12.2 and Hsp12.3 form tetramers and have no chaperone-like activity. AB - Four 12.2-12.6 kDa small heat-shock proteins (sHSPs) of Caenorhabditis elegans are the smallest known members of the sHSP family. They essentially comprise the characteristic C-terminal 'alpha-crystallin domain' of the sHSPs, having a very short N-terminal region, and lacking a C-terminal tail. Recombinant Hsp12.2 and 12.3 are characterized here. Far-UV CD spectra reveal, as for other sHSPs, predominantly a beta-sheet structure. By gel permeation and crosslinking, they are the first sHSPs shown to occur as tetramers, rather than forming the usual large multimeric complexes. Exceptionally, too, both appear devoid of in vitro chaperone-like abilities. This supports the notion that tetramers are the building blocks of sHSP complexes, and that higher multimer formation, mediated through the N-terminal domains, is a prerequisite for chaperone-like activity. PMID- 9744801 TI - The Ecl18kI restriction-modification system: cloning, expression, properties of the purified enzymes. AB - Ecl18kI is a type II restriction-modification system isolated from Enterobacter cloaceae 18kI strain. Genes encoding Ecl18kI methyltransferase (M.Ecl18kI) and Ecl18kI restriction endonuclease (R.Ecl18kI) have been cloned and expressed in Escherichia coli. These enzymes recognize the 5'.../CCNGG...3' sequence in DNA; M.Ecl18kI methylates the C5 carbon atom of the inner dC residue and R.Ecl18kI cuts DNA as shown by the arrow. The restriction endonuclease and the methyltransferase were purified from E. coli B834 [p18Ap1] cells to near homogeneity. The restriction endonuclease is present in the solution as a tetramer, while the methyltransferase is a monomer. The interactions of M.Ecl18kI and R.Ecl18kI with 1,2-dideoxy-D-ribofuranose containing DNA duplexes were investigated. The target base flipping-out mechanism is applicable in the case of M.Ecl18kI. Correct cleavage of the abasic substrates by R.Ecl18kI is accompanied by non-canonical hydrolysis of the modified strand. PMID- 9744802 TI - Free fatty acids regulate the uncoupling protein and alternative oxidase activities in plant mitochondria. AB - Two energy-dissipating systems, an alternative oxidase and an uncoupling protein, are known to exist in plant mitochondria. In tomato fruit mitochondria linoleic acid, a substrate for the uncoupling protein, inhibited the alternative oxidase sustained respiration and decreased the ADP/O ratio to the same value regardless of the level of alternative oxidase activity. Experiments with varying concentrations of linoleic acid have shown that inhibition of the alternative oxidase is more sensitive to the linoleic acid concentration than the uncoupling protein activation. It can be proposed that these dissipating systems work sequentially during the life of the plant cell, since a high level of free fatty acid-induced uncoupling protein activity excludes alternative oxidase activity. PMID- 9744803 TI - Thermally induced chain exchange of smooth muscle tropomyosin dimers studied by differential scanning calorimetry. AB - The thermal unfolding of duck gizzard tropomyosin dimers, alphabeta, alphaalpha, and betabeta, and of a 1:1 mixture of alphaalpha and betabeta homodimers was studied by differential scanning calorimetry (DSC). Both alphaalpha and betabeta homodimers demonstrated a broad thermal transition with maxima at 37.4 degrees C and 44.6 degrees C, respectively. However, a sharp cooperative thermal transition at 41.5 degrees C characteristic for alphabeta heterodimer appeared on the thermogram of the mixture of homodimers. The appearance of this transition was prevented by disulfide cross-linking of polypeptide chains in the homodimers. Thus, DSC studies clearly demonstrate formation of tropomyosin heterodimers during heating of the mixture of homodimers and in agreement with earlier published reports indicate thermally induced chain exchange between tropomyosin dimers. PMID- 9744804 TI - Functional characterization of ORCTL2--an organic cation transporter expressed in the renal proximal tubules. AB - Chromosome 11p15.5 harbors a gene or genes involved in Beckwith-Wiedemann syndrome that confer(s) susceptibility to Wilms' tumor, rhabdomyosarcoma, and hepatoblastoma. We have previously identified a transcript at 11p15.5 which encodes a putative membrane transport protein, designated organic cation transporter-like 2 (ORCTL2), that shares homology with tetracycline resistance proteins and bacterial multidrug resistance proteins. In this report, we have investigated the transport properties of ORCTL2 and show that this protein can confer resistance to chloroquine and quinidine when overexpressed in bacteria. Immunohistochemistry analyses performed with anti-ORCTL2 polyclonal antibodies on human renal sections indicate that ORCTL2 is localized on the apical membrane surface of the proximal tubules. These results suggest that ORCTL2 may play a role in the transport of chloroquine and quinidine related compounds in the kidney. PMID- 9744805 TI - A trypanosome oligopeptidase as a target for the trypanocidal agents pentamidine, diminazene and suramin. AB - African trypanosomes contain a cytosolic serine oligopeptidase, called OP-Tb, that is reversibly inhibited by the active principles of three of the five most commonly used trypanocidal drugs: pentamidine, diminazene and suramin. OP-Tb was inhibited by pentamidine in a competitive manner, and by suramin in a partial, non-competitive manner. The inhibition of OP-Tb by a variety of suramin analogues correlated with the trypanocidal efficacy of these analogues (P=0.03; by paired Student's t-test). Since intracellular (therapeutic) concentrations of pentamidine and suramin are reported to reach approximately 206Ki and 15Ki respectively, we suggest that these drugs may exert part of their trypanocidal activity through the inhibition of OP-Tb. PMID- 9744806 TI - Identification, functional expression and chromosomal localisation of a sustained human proton-gated cation channel. AB - Non-inactivating or slowly inactivating proton-gated cation channels are thought to play an important role in the perception of pain that accompanies tissue acidosis. We have identified a novel human proton-gated cation channel subunit that has biphasic desensitisation kinetics with both a rapidly inactivating Na+ selective and a sustained component. The protein shares 84% sequence identity with the proton-gated cation channel rASIC3 (rDRASIC) from rat sensory neurones. The biphasic desensitisation kinetics and the sequence homology suggest that this novel clone (hASIC3) is the human orthologue of rASIC3 (rDRASIC). While rASIC3 (rDRASIC) requires very acidic pH (pH < 4.5) for activation of the sustained current, the non-inactivating hASIC3 current starts to be activated when the pH decreases to below pH 6. hASIC3 is an acid sensor and might play an important role in the detection of lasting pH changes in human. We localised the hASIC3 gene to the human chromosome 7q35, 6.4 cRad telomeric from the microsatellite AFMA082XC9. PMID- 9744807 TI - Alteration of the ADP/ATP translocase isoform pattern improves ATP expenditure in developing rat liver mitochondria. AB - The expression of adenine nucleotide translocase isoforms (AAC) during perinatal development of the rat was studied by measuring mRNA transcript levels of AAC1 and AAC2 genes in liver, heart and brain tissue. In contrast to heart and brain, AAC1 mRNA is not present in adult liver tissue, but is transiently expressed around birth. AAC1 expression in liver did not respond to cold stress (examined with adult rats), therefore a possible involvement of AAC1 in the liver thermogenesis of newborns is excluded. Measurement of the [3H]ADP uptake by liver mitochondria revealed that the molecular activity of the AAC protein was significantly higher in mitochondria from 4-day-old neonates compared with adults. We suggest that the transient AAC1 gene expression in the perinatal liver helps to accommodate the mitochondrial ATP supply to the increased cytosolic ATP consumption initiated at birth. PMID- 9744808 TI - Apoptosis induced by modified ribonucleosides in human cell culture systems. AB - The in vitro modulation of apoptosis and cell proliferation by modified in comparison with non-modified ribonucleosides was investigated for the first time using peripheral blood lymphocytes, HL-60 cells and Caco-2 cells as human cell culture models. Modulating effects of several ribonucleosides were found in the range of 10(-7)-10(-3) mol/l. The following ribonucleosides induced significant apoptosis of HL-60 cells: adenosine, N6-dimethyladenosine, N6-(2-isopentenyl) adenosine, N2-dimethylguanosine. A significant apoptotic effect on PBL was found with N6-dimethyladenosine and N6-(2-isopentenyl)-adenosine. N6-Dimethyladenosine, N6-(2-isopentenyl)-adenosine and guanosine had a pronounced inhibitory effect on Caco-2 cell apoptosis. Regarding the known function of ribonucleosides as pathobiochemical marker molecules for cancer, the possibility of a selective apoptotic effect against malignant cells is discussed. PMID- 9744809 TI - Impairment of tRNA processing by point mutations in mitochondrial tRNA(Leu)(UUR) associated with mitochondrial diseases. AB - Several point mutations in mitochondrial tRNA genes have been linked to distinct clinical subgroups of mitochondrial diseases. A particularly large number of different mutations is found in the tRNA(Leu)(UUR) gene. We show that base substitutions at nucleotide position 3256, 3260, and 3271 of the mitochondrial genome, located in the D and anticodon stem of this tRNA, and mutation 3243 changing a base involved in a tertiary interaction, significantly impair the processing of the tRNA precursor in vitro. In correlation with other studies, our results suggest that inefficient processing of certain mutant variants of mitochondrial tRNA(Leu)(UUR) is a primary molecular impairment leading to mitochondrial dysfunction and consequently to disease. PMID- 9744810 TI - Dimeric and tetrameric forms of catalytically active transmembrane CD38 in transfected HeLa cells. AB - CD38, a type II transmembrane glycoprotein, behaves as a catalytically active transporter responsible for ectocellular generation of cyclic ADP-ribose (cADPR) from NAD+ and for subsequent influx of cADPR across membranes [Franco, L., Guida, L., Bruzzone, S., Zocchi, E., Usai, C. and De Flora, A. (1998) FASEB J. in press]. cADPR regulates intracellular calcium homeostasis by releasing calcium from responsive stores. The cADPR-transporting function of CD38 requires channel generating oligomeric forms of the protein rather than the 46 kDa monomers that have been described so far in CD38+ cells. Here we demonstrate that CD38, both in reconstituted proteoliposomes and in CD38-transfected HeLa cells, is a mixture of catalytically active monomers, homodimers and homotetramers. A soluble recombinant form of CD38 corresponding to its ectocellular region proved to be monomeric. Thus, association of native CD38 with either artificial or natural membranes seems to result in a reversible juxtaposition of monomers suitable to cADPR-transporting activity. PMID- 9744811 TI - Pyridoxal phosphate binding to wild type, W330F, and C298S mutants of Escherichia coli apotryptophanase: unraveling the cold inactivation. AB - The mechanism of pyridoxal phosphate (PLP) binding to apotryptophanase was investigated using stopped-flow kinetics with wild type (WT), W330F and C298S mutants. Based on the dependence of the rate constants on PLP concentrations for the fast and slow phases detected, two mechanistic schemes were proposed. For the WT and C298S mutant, the slow process is due to an isomerization of the aldimine complex after its formation, and not to the binding to an alternative conformation of the apoenzyme, which is the case proposed for the W330F mutant. It is suggested that during the cold inactivation process a conformational change precedes the aldimine bond cleavage. For the W330F apotryptophanase, another conformational change occurs subsequent to the aldimine bond cleavage. PMID- 9744812 TI - T4 DNA ligase synthesizes dinucleoside polyphosphates. AB - T4 DNA ligase (EC 6.5.1.1), one of the most widely used enzymes in genetic engineering, transfers AMP from the E-AMP complex to tripolyphosphate, ADP, ATP, GTP or dATP producing p4A, Ap3A, Ap4A, Ap4G and Ap4dA, respectively. Nicked DNA competes very effectively with GTP for the synthesis of Ap4G and, conversely, tripolyphosphate (or GTP) inhibits the ligation of DNA by the ligase. As T4 DNA ligase has similar requirements for ATP as the mammalian DNA ligase(s), the latter enzyme(s) could also synthesize dinucleoside polyphosphates. The present report may be related to the recent finding that human Fhit (fragile histidine triad) protein, encoded by the FHIT putative tumor suppressor gene, is a typical dinucleoside 5',5''-P1,P3-triphosphate (Ap3A) hydrolase (EC 3.6.1.29). PMID- 9744813 TI - Apoptosis-like, reversible changes in plasma membrane asymmetry and permeability, and transient modifications in mitochondrial membrane potential induced by curcumin in rat thymocytes. AB - Curcumin (diferuoylmethane) is a natural compound with anticarcinogenic activities which is able to exert either proapoptotic or antiapoptotic effects in different cell types. This paper focuses on the sequence and extent of primary events induced by curcumin, in comparison with those occurring during dexamethasone-induced apoptosis in rat thymocytes. It also presents annexin VI FITC as a new probe for studying membrane asymmetry. Curcumin readily penetrates into the cytoplasm, and is able to accumulate in membranous structures such as plasma membrane, endoplasmic reticulum and nuclear envelope. Curcumin-treated cells exhibit typical features of apoptotic cell death, including shrinkage, transient phosphatidylserine exposure, increased membrane permeability and decrease in mitochondrial membrane potential. However, nuclei morphology, DNA fragmentation, the extent and time-course of membrane changes are different from those observed during dexamethasone-induced apoptosis, suggesting that, despite many similarities, the mode of action and the events triggered by curcumin are different from those occurring during typical apoptosis. PMID- 9744814 TI - Protein phosphatase inhibitors and heat preconditioning prevent Hsp27 dephosphorylation, F-actin disruption and deterioration of morphology in ATP depleted endothelial cells. AB - The vascular endothelium response to ischemic depletion of ATP was studied in vitro. Endothelial cells (EC) cultured from human aorta or umbilical vein were incubated in a glucose-free medium containing CCCP or rotenone. Such blockade of energy metabolism caused a drop in ATP, destruction of actin filaments, morphological changes, and eventually disintegration of EC monolayer within 2-2.5 h. While ATP fell and F-actin collapsed, the 27-kDa heat shock protein (Hsp27) lost basal phosphorylation and became Triton X-100-insoluble forming granules inside the cell nuclei. Protein phosphatase (PP) inhibitors (okadaic acid, cantharidin, sodium orthovanadate) did not delay the ATP decrease in energy deprived EC but arrested both the alterations in the Hsp27 status and the changes for the worse in F-actin and cell morphology. Similarly, the Hsp27 dephosphorylation/insolubilization/granulation and the cytoskeletal and morphological disturbances resulting from lack of ATP were suppressed in heat preconditioned (thermotolerant) cultures, this effect being sensitive to quercetin, a blocker of Hsp induction. The longer preservation of the cytosolic pool of phosphorylated Hsp27 during ATP depletion in the PP inhibitor-treated or thermotolerant EC correlated with the acquired resistance of F-actin and morphology. These data suggest that PP inhibitors as well as heat-inducible Hsp(s) can protect ischemia-stressed cells by preventing the ATP loss-provoked protein dephosphorylation and breakdown of the actin cytoskeleton. PMID- 9744815 TI - NMR study on the impact of metal ion binding and deoxynucleotide substitution upon local structure and stability of a small ribozyme. AB - We have studied a very small ribozyme described earlier which requires the presence of soft metal ions like manganese or cadmium. It consists of only three uridines as ribozyme, cleaving the sequence 5'-GAAA-3' after the guanosine. We have set out to characterize the metal ion binding in this system by NMR spectroscopy and the impact of the ribose 2'-OH group of the cleavable nucleotide upon local structure. NMR results indicate a high degree of regularity and order in the pyrimidine-rich ribozyme strand, and high flexibility within the purine rich substrate. The guanosine 2'-hydroxy group adjacent to the cleavage site was found to have a profound effect upon the structure, apparently destabilizing a stacked arrangement. Metal ions were found to bind in a rather unspecific way, however, in the presence of higher amounts of divalent ions a preference in the vicinity of the cleavage site could be observed. 113Cd NMR spectra suggest a specific binding of Cd2+ ions to the RNA. PMID- 9744816 TI - A comparative differential scanning calorimetric study of tobacco mosaic virus and of its coat protein ts mutant. AB - The differential scanning calorimetry (DSC) 'melting curves' for virions and coat proteins (CP) of wild-type tobacco mosaic virus (strain U1) and for its CP ts mutant ts21-66 were measured. Strain U1 and ts21-66 mutant (two amino acid substitutions in CP: 121 --> T and D66 --> G) differ in the type of symptoms they induce on some host plants. It was observed that CP subunits of both U1 and ts21 66 at pH 8.0, in the form of small (3-4S) aggregates, possess much lower thermal stability than in the virions. Assembly into the virus particles resulted in a DSC melting temperature increase from 41 to 72 degrees C for U1 and from 38 to 72 degrees C for ts21-66 CP. In the RNA-free helical virus-like protein assemblies U1 and ts21-66 CP subunits had a thermal stability intermediate between those in 3-4S aggregates and in the virions. ts21-66 helical protein displayed a somewhat lower thermal stability than U1. PMID- 9744817 TI - Complex regulation of multiple cytohesin-like genes in murine tissues and cells. AB - Three cytohesin-like cDNA molecules were isolated from murine ES cell derived embryoid bodies. The genomic structure of one of the three, CLM2, has been determined and transcriptional variants of each were isolated from a mouse brain cDNA library. The relative expression patterns of CLM1, 2, 3 and their transcriptional alternatives were determined by RT-PCR, nucleotide sequencing and RNA blotting. Their broad distribution and cell and tissue specific expression patterns suggest complex regulation during development and in the adult. PMID- 9744818 TI - Corpeptins, new bioactive lipodepsipeptides from cultures of Pseudomonas corrugata. AB - The structure of the corpeptins, bioactive lipodepsipeptides produced in culture by Pseudomonas corrugata, the causal agent of tomato pith necrosis, has been determined. The combined use of FAB-mass spectrometry, NMR spectroscopy and chemical procedures has allowed us to assign the following primary structure to the peptide moiety: Dhb-Pro-Ala-Ala-Ala-Val-Val-Dhb-Hse-Val-alle-Dhp-Ala-Ala-Ala Val-D hb-aThr-Ala-Dab-Ser-Ile with the terminal carboxy group closing a macrocyclic ring on the hydroxy group of the allo-threonine residue. The N terminus is in turn acylated by 3-hydroxydecanoate in corpeptin A and by cis-3 hydroxy-5-dodecenoate in corpeptin B. Some preliminary data on the biological activity of corpeptins are included. PMID- 9744819 TI - Function and picosecond dynamics of bacteriorhodopsin in purple membrane at different lipidation and hydration. AB - By neutron scattering experiments and time-resolved absorption spectroscopy we have investigated picosecond equilibrium fluctuations and the kinetics of the photocycle of bacteriorhodopsin (BR) in the purple membrane (PM). Natural PM samples composed of 75% BR (w/w) and 25% lipid (w/w) as well as delipidated PM having only 5% lipid (w/w) were measured at different levels of hydration. We observed a reduced 'flexibility', due to a diminished weight of stochastic large amplitude motions occurring in the delipidated PM as compared to the natural PM. This effect is more pronounced for wet samples, indicating the importance of lipid hydration for protein dynamics. The reduced flexibility is accompanied by significantly larger time constants describing the decay of the M-intermediate. Therefore, a correlation between the dynamical behavior of the protein-lipid complex and BR function emerges. PMID- 9744820 TI - GTP analogue hydrolysis by the Gs protein: implication for the role of catalytic glutamine in the GTPase reaction. AB - Hydrolysis of GTP, bound to members of the G-protein superfamily, terminates their downstream signaling activity. A conserved glutamine serves a critical role in this pivotal guanosine triphosphatase (GTPase) reaction. However, the role of the catalytic glutamine in GTP hydrolysis is still not well understood. We have employed substrate-assisted catalysis to probe the catalytic mechanism of Gs alpha using GTP analogues. These GTP analogues, each having different functional groups, were designed to support or refute particular putative GTPase mechanisms. We have found that a hydrogen donor group, in close proximity to the gamma phosphate of GTP, is necessary and sufficient to substitute for the function of the catalytic glutamine in the GTPase reaction. PMID- 9744822 TI - Current advances in the management of stroke. PMID- 9744821 TI - Production of parathyroid hormone-related peptide by synovial fibroblasts in human osteoarthritis. AB - Synovial fibroblasts from patients with osteoarthritis in culture produced parathyroid hormone-related peptide (PTHrP) on treatment with phorbol ester (TPA) in a dose- and time-dependent manner. The levels of PTHrP immunoreactivity in the conditioned medium of synovial fibroblast cultures were measured using specific PTHrP antibody. The maximum production was obtained at a concentration of 10(-8) M and 24 h after TPA treatment. But sensitivity to TPA of synovial fibroblasts differed among four patients from slight to marked. PTHrP production was also induced with inflammatory cytokines, such as 1 ng/ml of IL-1alpha, IL-1beta, IL-6 and TNF-alpha, and 10(-6) M prostaglandin E2, after 24 h treatment. The expression of PTHrP was confirmed by reverse-transcriptase polymerase chain reaction. Since the synovial fibroblasts isolated from osteoarthritic patients produce high levels of IL-6 and IL-8, typical cytokines produced in synovial fibroblasts, production of PTHrP may provide new insight into the pathophysiology of joint disorder. PMID- 9744823 TI - Leukoaraiosis and vascular dementia. AB - The emergence of sensitive techniques for brain imaging has drawn attention to the occurrence of diffuse or multifocal changes affecting the cerebral white matter. The white matter changes are usually termed periventricular leukoencephalopathy, or leukoaraiosis. Microscopic studies of affected areas in the deep white matter have shown mostly demyelination, reactive gliosis, and arteriolosclerosis, proportional to the degree of radiologic changes. Yet, many other disease conditions need to be ruled out. Risk factors for ischemic leukoaraiosis include arterial hypertension, a history of stroke, and age. In the hereditary disorder CADASIL (cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy), severe white matter changes occur in the absence of hypertension. In "ordinary" cases of leukoaraiosis, genetic factors might similarly determine the effect of risk factors on the aging brain and might explain, for example, why not all patients with severe hypertension develop leukoaraiosis. Not surprisingly, diffuse demyelination affects cognitive function. Although reduced speed of mental processes is the most characteristic sign, attention, concentration, and verbal and visual memory are also affected. Most importantly, less severe forms of cognitive impairment represent a silent and perhaps largely preventable epidemic among aged or even middle-aged subjects. They live independently, but mentally they perform on a level well below their previous capacities. Although being "a bit odd" does not lead to hospital admissions, it seriously affects quality of life of a large part of the community. Moderate grades of leukoaraiosis constitute a major public health problem and deserve the attention of the scientific community. PMID- 9744824 TI - The role of platelets in ischemic stroke. AB - Platelets have assumed a role in the development of focal cerebral ischemia by virtue of their participation in thromboemboli that may initiate stroke symptoms. Platelets are one component of the blood-vascular axis responsible for preventing hemorrhage. Activated platelets initiate hemostatic plug formation and provide a scaffolding for coagulation activation. Platelets are activated by a number of stimuli, such as exposure of the vascular subendothelium, fibrin deposition, and abnormal surfaces, e.g., atheromata. A number of observations, including the appearance of platelet thrombi on atheromata in situ, indicate that platelet physiology is relevant to stroke. In addition, certain antiplatelet agents (e.g., aspirin) significantly reduce the incidence of ischemic stroke after initial transient ischemic attacks. Aspirin, the combination of aspirin and dipyridamole, and ticlopidine have all been shown to be useful in reducing the frequency of secondary stroke events. Clopidogrel has been shown to reduce the frequency of secondary vascular ischemic events when stroke, myocardial infarction, and peripheral arterial disease are considered together. Unfortunately, all antithrombotic agents carry a potential risk for inducing symptomatic intracerebral hemorrhage during ischemic stroke. The mechanism by which this may happen with antiplatelet agents has not yet been determined. As in other areas of stroke treatment, it is the balance between efficacy in reduction of symptomatic thrombotic events and the risk for hemorrhage that will define benefit. PMID- 9744825 TI - Low-dose and high-dose acetylsalicylic acid, with and without dipyridamole: a review of clinical trial results. AB - Publication of the results of the second European Stroke Prevention Study (ESPS 2) provided the incentive for an update of the meta-analyses of aspirin and dipyridamole in the secondary prevention of stroke. After review of published randomized trials of prolonged treatment with aspirin, dipyridamole, or their combination in patients with a history of stroke or transient ischemic attack (TIA), data on the occurrence of stroke, myocardial infarction, and vascular death were used to calculate overall relative risk reductions. The relative risk reduction for aspirin versus placebo was 13%. The same relative risk reduction was found in separate meta-analyses of trials with high (1,000-1,500 mg), medium (250-500 mg), and low (50-100 mg) doses of aspirin. Trials in which different doses were compared showed no difference in the occurrence of vascular events. The addition of dipyridamole to low-dose aspirin further reduced the risk for vascular events by 15%. We conclude from current trials that low-dose aspirin alone reduced the risk of vascular events in patients with prior stroke or TIA by 13%. There is no evidence of a dose-effect relationship. An additional reduction of the risk by 15% can be obtained by adding dipyridamole to aspirin. The overall evidence for the relative effects of the combination of dipyridamole and aspirin versus aspirin alone or placebo is highly consistent. The clinical evidence now favors the two agents in combination over aspirin alone. PMID- 9744826 TI - Dipyridamole trials in stroke prevention. AB - Research on the benefits of aspirin combined with other antiplatelet regimens for stroke prevention has yielded inconclusive results. Early trials of aspirin plus dipyridamole (DP) were unable to detect a significant benefit for combination therapy over aspirin alone, although they clearly demonstrated the value of combination therapy compared with placebo. Early trials such as the AICLA (Accidents ischemiques cerebraux lies a l'atherosclerose) trial and the American Canadian Cooperative Study lacked the statistical power to detect differences in the benefit of combination versus monotherapy because of the small number of events in each treatment group. The Antiplatelet Collaboration, in its meta analysis published in 1994, also failed to detect a significant difference between the benefit of aspirin plus DP and that of aspirin alone for the combined end point of stroke, myocardial infarction, and vascular death. The large European Stroke Prevention Study 2 (ESPS-2) trial recently provided evidence that aspirin plus DP does lead to a significantly greater reduction than aspirin alone. The 6,602 patient trial randomized patients into four treatment groups: aspirin (50 mg daily) plus sustained-release DP (400 mg daily), aspirin alone, DP alone, or placebo. The trial found that low-dose aspirin plus DP more than doubled the reduction in stroke risk achieved with aspirin alone, a 37% risk reduction for the combination versus 18.1% for aspirin alone. The results also suggest that the effects of aspirin and DP are additive. PMID- 9744827 TI - Anticoagulation for prevention of stroke. AB - One of the important recent advances in stroke prevention is the demonstration that warfarin can substantially reduce the risk for stroke in patients with atrial fibrillation (AF). On average, patients with AF have a stroke risk of 4.5% per year. Anticoagulation reduces this to around 1.5% per year, a 70% relative risk reduction. The presence of additional risk factors, such as a recent stroke or transient ischemic attack, hypertension (particularly systolic hypertension), congestive heart failure, or diabetes, greatly increases stroke risk. Patients with any of these risk factors have a stroke risk of 8% per year or more. In contrast, patients under age 75 with none of these risk factors have a low risk for stroke (around 1% per year) when treated with aspirin. This risk stratification may help in identifying which patients with AF benefit most from anticoagulation. Anticoagulation has also been shown to prevent stroke in patients with other cardioembolic sources, including acute anterior wall myocardial infarction (particularly with echocardiographic evidence of thrombus), prosthetic heart valves, and dilated cardiomyopathies. PMID- 9744828 TI - Cardiac evaluation of stroke patients. AB - There are two potential purposes for cardiac evaluation in patients with cerebrovascular disease: to identify possible cardioembolic pathophysiology for ischemic symptoms and to identify concomitant coronary artery disease. Both have important implications for patient prognosis and treatment, and testing therefore appears to be warranted. On the other hand, the cost conservation movement in medicine dictates that physicians limit unnecessary, costly, possibly risky testing when the diagnostic yield is low. For example, the overall yield of cardiac testing in "usual stroke patients" who have no suggestive history or findings on examination, chest X-ray, or electrocardiogram is less than 10% and may not be indicated routinely. Conversely, young patients with stroke of unknown cause are likely to benefit from aggressive cardiac testing. Many reported series and clinical trials have demonstrated that patients with cerebrovascular disease are more likely to die in follow-up from cardiovascular than from cerebrovascular causes. This risk is best defined and may be highest in patients with carotid disease, in whom the 5-year cardiac mortality rate may be as high as 40 to 50%. Studies have shown that such patients are also likely to have abnormal tests for cardiac ischemia, even when a history of cardiovascular events or symptoms or electrocardiographic abnormalities are lacking. These results, combined with further investigations into which cerebrovascular patients are at highest risk for cardiovascular disease and what testing best identifies underlying, treatable cardiovascular disease, are needed to direct the care and improve the cardiovascular prognosis of patients with cerebrovascular disease. PMID- 9744829 TI - Identifying patient populations at high risk for stroke. AB - Although the treatment of acute ischemic stroke has improved, the greatest reductions in stroke mortality and morbidity may possibly be achieved through more effective prevention strategies. Toward this goal, risk factor profiles for initial and recurrent stroke have been identified through longitudinal epidemiologic studies. Nonmodifiable risk markers for initial ischemic stroke include age, sex, family history, and race/ethnicity. Modifiable risk factors for first ischemic stroke include hypertension, cardiac disease (particularly atrial fibrillation), diabetes, hyperlipidemia, cigarette smoking, alcohol abuse, physical inactivity, asymptomatic carotid stenosis, and transient ischemic attack. As improved acute treatments increase survival after a first stroke, the threat of increased morbidity from stroke recurrence will have greater significance. The risk and specific determinants of early and late stroke recurrence are the subject of ongoing investigations. Age, stroke syndrome, hypertension, cardiac disease (particularly congestive heart failure), hyperglycemia, and alcohol abuse have been identified as predictors of late stroke recurrence. Now that many risk factors are established, greater emphasis should be placed on identifying high stroke-risk patient populations for intensive risk factor modification and antithrombotic treatments. Better understanding and management of stroke risk factors will undoubtedly improve our ability to prevent first and recurrent ischemic stroke. PMID- 9744830 TI - The value of stroke prevention and treatment. AB - An emerging issue in stroke prevention and stroke treatment is that therapies shown to be effective in clinical trials are not being used or are being used suboptimally. One of many explanations relates to cost, ranging from concerns about inappropriate use of societal resources to more immediate concerns of organizations and individuals about their financial well-being. Assessing the cost impact of a new therapy, and thus the financial incentives faced by decision makers, is crucial to understanding and influencing real-world treatment decisions. Assessing cost in the context of health benefits is the object of cost effectiveness analysis. A cost-effectiveness analysis typically compares the new treatment with a less efficacious yet less costly alternative. This article provides a brief overview of the cost implications of stroke and stroke-related treatments. The example of stroke prevention is used to illustrate how to calculate an incremental cost-effectiveness ratio and help clarify what makes a treatment an especially good value. Because stroke is tremendously expensive and because severe strokes are especially expensive, treatments to prevent stroke and to diminish stroke disability are likely to be an excellent value. PMID- 9744831 TI - What have we learned from recent antiplatelet trials? AB - Aspirin's benefit in preventing vascular outcomes is well established. It reduces the relative risk for stroke, myocardial infarction, and vascular death by about 25% compared with placebo. Almost 10 years ago we learned that ticlopidine is more effective than aspirin (about 12% relative risk reduction for stroke or death). However, ticlopidine has important adverse effects. In 1996, the Clopidogrel versus Aspirin in Patients at Risk of Ischemic Events (CAPRIE) trial showed that clopidogrel, a new thienopyridine similar to ticlopidine, is also more effective than aspirin (by a similar amount) and is as safe as aspirin. Also in 1996, the European Stroke Prevention Study 2 (ESPS-2) showed that dipyridamole alone prevents stroke and that when combined with aspirin it is more effective, probably comparable to ticlopidine and clopidogrel. Dipyridamole combined with aspirin reduced the relative risk for stroke or death by about 13% compared with aspirin alone. Both clopidogrel and dipyridamole are safe but will cost more than aspirin. Aspirin also appears beneficial for acute stroke treatment. The Chinese Acute Stroke Trial (CAST) and the International Stroke Trial (IST) demonstrated that aspirin given at the time of an acute ischemic stroke reduces the risk for early death (about 5 less/1,000 treated), recurrence or death (about 10 less/1,000 treated), and dependence (about 5 less/1,000 treated). Overall, the benefits of aspirin in acute stroke treatment and stroke prevention are definite but modest. Combination therapy with antiplatelet agents that act through different mechanisms is a promising way to maximize the benefits of antiplatelet treatment. PMID- 9744832 TI - Insulin, blood glucose levels, and ischemic brain damage. AB - Intensive efforts are presently directed toward developing pharmacologic therapy to protect the ischemic brain. Preclinical data from animal models indicate that insulin, already available for human use, may reduce damage in both global and focal ischemia. Two kinds of mechanisms may be involved: one in which insulin interacts directly with brain tissue and one in which insulin acts indirectly by reducing peripheral blood glucose levels. Animal data indicate that part of the former, direct mechanism is mediated by insulin-like growth factor-1 receptors. The direct effect appears to predominate in global ischemia. In focal ischemia, unlike global ischemia, the effect of insulin is predominantly via peripheral hypoglycemia, because neuroprotection is largely annulled by co-administration of glucose. The two clinical counterparts of global and focal ischemic models are, respectively, cardiac arrest encephalopathy and focal ischemic stroke. Insulin use in both of these clinical situations could be evaluated in clinical trials that attempt to reduce ischemic brain damage, because insulin has a long and safe history of human use in diabetes treatment. PMID- 9744833 TI - Ischemic penumbra: the therapeutic window. AB - The concept of ischemic penumbra has extended our understanding of the focal ischemic process and in many ways has revitalized our research. Simply stated, the penumbra is the part of the brain that is sandwiched between brain regions committed to die and those that receive enough blood to communicate. Therefore, it is ischemic brain tissue that has just enough energy to survive for a short time but not enough to communicate and function. The life expectancy of the penumbra is short. Although the penumbra is an elegant concept, in practice it has been a difficult one to exploit. For one thing, it is unstable in both time and space. Depending on the severity and the duration of the focal ischemia, it may be anywhere in the ischemic brain. We believe that nimodipine binding experiments have taught us a great deal about the ischemic penumbra. Second, cells in the penumbra may die not by necrosis but by apoptosis. If that is true, then our concepts about the benign transient ischemic attack may need revision. Third, the penumbra may regain its ability to survive not only through reperfusion but also by interrupting the process of commitment to apoptosis. PMID- 9744834 TI - Diffusion-weighted MRI for evaluation of acute stroke. AB - Diffusion-weighted imaging (DWI) is a new magnetic resonance imaging technique that detects the tiny random movements of water molecules (diffusion) in tissues. This technique allows a map of the average apparent diffusion coefficient (ADC) to be calculated. Shortly after the onset of an ischemic stroke, the ADC of brain tissue is significantly reduced because of cytotoxic edema. Over several days, the rapid initial drop in ADC is followed by a return to "pseudonormal" values at approximately 1 week. Subsequently, elevated ADC values are seen at chronic time points. DWI is remarkably sensitive in detecting and localizing acute ischemic brain lesions and allows differentiation of acute regions of ischemia from chronic infarcts. Recent studies have shown a high correlation between the volume of early DWI lesions and clinical neurologic outcome. In addition, the volume of the early DWI lesion correlates well with final infarct volume as measured by T2 weighted imaging. Therefore, this technique may facilitate optimal selection of patients for new medical therapies for stroke and may provide a highly sensitive technique for evaluating the efficacy of new treatments. PMID- 9744835 TI - Effect of training in reading CT scans on patient selection for ECASS II. AB - Anticipating that patients with large ischemic lesions exceeding one-third of the middle cerebral artery (MCA) territory and detected on CT scans within 6 hours of stroke onset will not benefit from thrombolysis, we trained each participant of the second European Cooperative Acute Stroke Study (ECASS II) in the recognition of early ischemic lesions. Participants (n=532; neurologists, radiologists, neuroradiologists) were tested before and after each 4-hour CT reading training course. We asked the participants to estimate the extent of acute ischemic lesions on 10 CT scans, which we presented for 90 seconds without clinical information. Two sets of 10 CT scans each (A and B) were alternatively presented to each group, so that 254 participants evaluated set A before the training and 278 participants evaluated set B. We compared the numbers of correct estimates, underestimations, and overestimations before and after the course for each participant. The person who taught all courses (RvK) provided the reference estimates. We found that training significantly increased the number of correct estimates (p < 0.0001). Subsequently, we studied the incidences of large infarctions and parenchymal hemorrhages in the ECASS II population. In comparison with ECASS I investigators, the local investigators of ECASS II reduced the number of falsely included patients to an extent similar to that of the training courses. More remarkably, among the ECASS II patients, the proportion of patients with large infarctions or parenchymal hemorrhages was reduced to 50%. Careful CT reading may have contributed to this result. PMID- 9744836 TI - tPA in acute ischemic stroke: United States experience and issues for the future. AB - The approval of tissue plasminogen activator (tPA) for treatment of patients with ischemic stroke in the United States marked the first therapy proven to reverse or limit the effects of an acute stroke. Despite this approval and the lack of an alternative therapy, the use of tPA in stroke has been quite low. Several explanations for this underutilization have been identified, including lack of patient awareness, potential complications, infrastructure deficiencies, and physician concerns. This article explores these issues and suggests strategies for improving the use of tPA as an acute therapy in stroke. PMID- 9744837 TI - Heparin and heparinoids in stroke. AB - Anticoagulation with heparin has a valuable place in prevention and management of deep venous thrombosis. However, the benefit of heparin in acute ischemic stroke and transient ischemic attack remains unclear despite its widespread use for these indications. Heparin also carries several risks, including unpredictable anticoagulation effects, bleeding, and thrombocytopenia. Low-molecular-weight heparins (LMWHs) and heparinoids have several advantages over heparin, such as higher bioavailability, more predictable anticoagulant effects, and less interaction with platelets. Heparin, LMWHs, and heparinoids have been studied in acute ischemic stroke with variable results. Of three recent, large, controlled clinical trials, only one documented a net benefit of treatment. Fewer patients treated with an LMWH within 48 hours of stroke were dead or disabled at 6 months compared with placebo-treated patients. The largest randomized clinical trial of heparin in acute stroke (the International Stroke Trial) showed that heparin was associated with a significant excess in bleeding complications but no clinical benefit at 6 months. Interim analysis of the TOAST (Trial of ORG 10172 in Acute Stroke Treatment) study also showed an excess number of bleeding complications in the treated group without a corresponding benefit on stroke outcome at 3 months. Therefore, although heparin, LMWHs, and heparinoids continue to be used in the management of patients with acute ischemic stroke, their value in recurrent stroke prevention and in the treatment of stroke-in-progress remains unsettled. Ongoing studies may help to clarify the use of LMWHs and heparinoids in these patients. PMID- 9744838 TI - Clinical trials in acute stroke: why have they not been successful? AB - Modeling of focal cerebral ischemia seeks to understand mechanisms of injury and to test agents as potential stroke therapies. However, modeling has been singularly unpredictable in ischemic cerebrovascular disease for a number of reasons related to the incompletely understood pathophysiology of ischemic stroke and to the characteristics of models prepared to mimic the clinical condition. The development of models of focal cerebral ischemia must take into account known species differences and idiosyncrasies, underlying vascular disease processes, the nature of thrombotic processes, cellular reactivities, the presence of co stimulation (e.g., inflammation), the characteristics of immunologicals and reporter molecules, the coincident use of other pharmacologic modifiers (e.g., anesthesia), and stress. These elements are also potential contributors to cerebral tissue injury and its assessment but may affect other species differentially. On the other hand, study design issues have been shown to be particularly relevant to limiting development of some agents for clinical stroke treatment. Experience from experimental and clinical work on vascular active approaches (e.g., plasminogen activators) suggests that active dialogue regarding the relationships between clinical outcomes and outcomes in appropriate animal models is necessary. Success appears more likely when the model more closely matches the known human pathophysiology and the interventions applied in the models are definitely characterized in that species. Rather than moving directly to interventional studies in humans, the use of several appropriate animal models is encouraged where those models exist. Where not, careful consideration of study design and the biology of the disorder is a prerequisite. PMID- 9744839 TI - The role of inflammation after acute stroke: utility of pursuing anti-adhesion molecule therapy. AB - A growing body of evidence, primarily from animal models of cerebral ischemia and preliminary human studies, indicates that inflammatory mechanisms contribute to secondary neuronal injury after acute cerebral ischemia. Ischemia followed by reperfusion rapidly leads to the expression of inflammatory cytokines, particularly tumor necrosis factor-alpha and interleukin-1beta, which stimulate a complex cascade of events involving local endothelial cells, neurons, astrocytes, and perivascular cells. A secondary response includes the release of other cytokines, an increase in components of the coagulation system, an upregulation of cell adhesion molecule expression, and changes in the expression of components of the immune response. The net effect of these events is transformation of the local endothelium to a prothrombotic/proinflammatory state and induction of leukocyte migration to the site of injury. A number of studies have shown that leukocyte migration occurs within hours of reperfusion. Leukocytes accumulate in the injured region, where they cause tissue injury by several mechanisms, including occlusion of microvasculature, generation of oxygen free radicals, release of cytotoxic enzymes, alteration of vasomotor reactivity, and increase in cytokine and chemoattractant release. Monoclonal antibodies against leukocyte adhesion molecules have been shown to reduce infarct volume in animal models of ischemia-reperfusion. However, this treatment failed to show benefit in the Enlimomab Acute Stroke Trial. A number of factors may complicate the use of antibody directed adhesion molecule blockade in acute stroke and will be discussed in this article. Overall, an increased understanding of inflammatory and immunologic mechanisms still offers great potential for reducing acute stroke injury. PMID- 9744840 TI - Surgery for intracerebral hemorrhage. AB - Intracerebral hemorrhage (ICH) represents 8 to 15% of all strokes in the United States and 20 to 30% of all strokes in Japan and China. Although ICH represents a relatively small fraction of total strokes, it is a formidable disease, with a 30 day mortality rate two- to sixfold higher than that for ischemic stroke. Furthermore, it is a major cause of disability, with only 20% of patients becoming independent at 6 months. The most common risk factors for ICH are age, hypertension, and amyloid angiopathy, which are associated with damage to and weakening of the arterial/arteriolar wall leading to vessel rupture. The pathology is a dynamic one that continues to evolve over the first few days after onset. In 20 to 30% of ICH, clot volume increases over the first 24 hours and is generally associated with neurologic worsening. The final outcome from ICH is related not only to clot volume, compression, and destruction but also to potential neurotoxicity from the blood degradation products and associated neuronal ischemia. The treatment of ICH has been one of the most controversial and least well-studied areas from a medical or surgical perspective. Surgical treatment has evolved over the years and can be grouped into open and stereotactically guided surgery for hematoma evacuation. Seven thousand operations per year are performed in the United States for hematoma evacuation, although this approach has not been adequately investigated. Adjuvant medical therapies with neuroprotective agents require further investigation and may potentially have additive benefits. PMID- 9744841 TI - Smoking cessation treatment for substance misusers in early recovery: a review of the literature and recommendations for practice. AB - Recent surveys have documented the very high incidence of smoking among treatment populations of alcohol and other drug misusers. While the health risks of smoking are well-documented in the literature, addictions professionals have traditionally been reluctant to address the problem of nicotine dependence with their clients. Recently, researchers have begun to investigate the impact of smoking cessation treatment on substance misusers who are also nicotine dependent. The purpose of this paper is to provide addictions treatment professionals with an overview of the research in this area and to highlight gaps in the knowledge base. In addition, this paper will review recent developments in the treatment of nicotine dependence and discuss their applicability to nicotine dependent persons who are in treatment for the misuse of alcohol or another drug. PMID- 9744843 TI - Primary socialization theory: culture, ethnicity, and cultural identification. The links between culture and substance use. IV. AB - Ethnicity, perceived membership in a cultural group, and cultural identification, the strength of one's affiliation with a group, develop primarily through interactions with the primary socialization sources, the family, the school, and peer clusters. Cultural norms for substance use are also transmitted as part of these interactions. Substance use differs across cultures; in different cultures some forms of substance use are culturally required, others are tolerated, and others are sanctioned. Ethnicity and cultural identification, therefore, should relate to substance use. However, primary socialization theory indicates that simple relationships are not likely to be found for a number of reasons: 1) All members of an ethnic group do not have the same level of cultural identification and may not, therefore, have the same conformance to substance use norms. 2) Primary socialization,sources are embedded in subcultures, and subcultures have norms that may differ from those of the larger ethnic group. 3) The individual may experience and report differing levels of cultural identification and different substance use norms in different social contexts. 4) For an individual, ethnicity and cultural identification may derive from different primary socialization sources than drug use norms. PMID- 9744842 TI - Social network ties, self-efficacy, and condom use among women who use crack cocaine: a pilot study. AB - So far, attempts to change the sexual risk behavior of women who use crack cocaine have been less successful than efforts to change the needle risk behavior of injection drug users. Two theoretical areas that have shown some success in predicting behavior change among of out-of-treatment drug users are Bandura's social cognitive theory (self-efficacy theory) and social network theory. According to Bandura, social networks are important sources of social support, and social support is vital to self-efficacy. Social network research also indicates that close bonds with network members may be a protective factor independently of self-efficacy. In order to test the feasibility of collecting such data, a pilot study was conducted with 60 women who used crack cocaine and who were not in treatment. Results of Pearson product-moment correlations indicated that self-efficacy (.7230) and number of very strong ties (.31252994) were positively correlated with condom use for women in the sample. In addition, the number of very strong ties (.3142) was significantly, if modestly, correlated with self-efficacy. Self-efficacy was PMID- 9744844 TI - Patterns of drug use among nonmetropolitan and rural adults. AB - This article examines illegal drug use among adults living in nonmetropolitan and rural areas of the United States using data from the National Household Survey on Drug Abuse. Subjects were classified into three categories by residence: nonmetropolitan-urban, metropolitan-rural, and nonmetropolitan-rural. Respondents indicate about 10% of adults were current users of marijuana or other illegal drugs. Discriminant analysis was used to examine differences among groups of individuals classified as current users, past users, and nonusers. For both marijuana and other illegal drugs, the variables that accounted most for group differences were age, marital, status, employment status, occupation, and income. Only minor differences in drug use were exhibited across the three residential categories. It is recommended that future research on the rural and nonmetropolitan adult population incorporate both structural level measures of socioeconomic and demographic characteristics of localities, and individual level measures of peer influence, work stress, family factors, and psychosocial characteristics. PMID- 9744845 TI - Primary prevention of alcohol misuse: overview and annotated bibliography. AB - Following an overview of conceptual and methodological issues related to alcohol misuse primary prevention and a brief discussion of the most frequently employed primary prevention strategies, a comprehensive annotated bibliography of the alcohol misuse primary prevention literature is presented. Several benefits of presenting detailed annotations, such as allowing readers to (a) examine the various prevention program components, (b) identify the presence or absence of methodological shortcomings, (c) identify whether or not high-risk groups were included as program participants, and (d) evaluate the feasibility of program implementation, are also highlighted. The present article complements previous reviews which have often mixed together the findings of primary and secondary prevention studies and which have typically reported the effects of prevention programming on multiple substance misuse outcomes simultaneously. The practice of simultaneously reporting multiple substance misuse outcomes has made it difficult to interpret the specific effects that primary prevention programs have had on alcohol misuse per se. PMID- 9744846 TI - Effects of interview mode on bias in survey measurements of drug use: do respondent characteristics make a difference? AB - Three recent empirical studies have provided strong evidence that self administered questionnaires (SAQs), compared with interviewer questioning, substantially improve the reporting of drug use in population surveys. Specifically, SAQs appear to diminish underreporting bias. Two of these studies previously reported that this effect of interview mode varied significantly across gender, race/ethnicity, and age. Data from a randomized experiment embedded in the 1990 National Household Survey of Drug Abuse (NHSDA) field test were reanalyzed to test for those interaction effects. To better replicate prior studies, the NHSDA field test sample was restricted to people ages 18 to 45 (N = 1,877). The results of our statistical analyses generally replicated the finding of a main effect of SAQs on the reporting of drug use. However, only weak evidence was found to support the hypothesis that the advantage of SAQs varies substantially by the gender, race/ethnicity, or age of the respondent. PMID- 9744847 TI - Ethanol and food pellet self-administration by baboons. AB - During daily 23-h sessions, baboons had concurrent access to food pellets and an oral ethanol/dextrose solution. The effect of increasing the fixed-ratio or "cost" for pellets on pellet and fluid intake was examined when baboons had access to 2%, 4%, or 8% (w/v) ethanol. Increasing the response requirement for a pellet decreased pellet intake. The rate of decrease in pellet intake with increasing pellet cost was unaffected by the availability of ethanol solutions, which were either self-administered or given in investigator-planned doses. Increasing the response cost for pellets significantly increased self administration of 4% ethanol. The effect of increasing the cost for fluid on fluid and pellet intake was examined when baboons had access to vehicle, 4% or 8% (w/v) ethanol. Although the total daily number of fluid deliveries was significantly greater when 4% ethanol was available, compared to vehicle, increasing the cost for a fluid delivery to 32 responses and above decreased intake of all three fluids similarly. Increasing the cost of ethanol did not affect food intake. PMID- 9744848 TI - Acute effect of ethanol on lens cation homeostasis. AB - This study examined the effect of ethanol on the calcium homeostasis of the bovine lens. After acute exposure of the whole lens to physiologically related ethanol concentration, the calcium content of the lens cortex increased from 0.345 +/- 0.075 to 0.476 +/- 0.047 micromol/g (p < 0.05). In contrast, other cation levels such as sodium, potassium, and magnesium did not change. In the study of the lens calcium transport, ethanol caused an increase in the calcium permeability of the lens lipid membrane by about 12% at 30 mM ethanol. Ethanol did not alter the calcium pump activity at ethanol concentration up to 400 mM. Above 600 mM ethanol, the calcium pump was almost completely inhibited. It has been suggested that moderate to heavy alcohol consumption is a risk factor for cataracts. This study indicates that acute ethanol exposure can cause a loss in the lens calcium homeostasis, which maybe one of the cellular mechanisms to contribute to the cataract development in the alcoholic individual. PMID- 9744849 TI - Magnesium levels in alcohol-treated rodents using different consumption paradigms. AB - To develop an animal model system that examines magnesium (Mg) deficiency associated with chronic alcohol consumption, we tried to reproduce a Mg-deficient state, caused by alcohol consumption, in rats using different alcohol consumption experimental designs. Serum and bone samples were collected from 2-day binge (high BACs achieved by intubating a 5% ethanol solution 2 consecutive days/week), 5-day binge, moderate drinking, and adolescent (4-week-old rats, equivalent to late teen/early adult humans) alcohol consumption projects. Mg content was measured using color spectrophotometry. Alcohol-fed animals consumed a liquid diet containing 0.38 g/kg/day ethanol in the moderate project and 35% ethanol derived calories in the adolescent drinking project. Animals in the two binge drinking projects were intubated with 12 g/kg/day of ethanol in a 5% solution. When looked at acutely and chronically, no consistent deficiencies in Mg were seen. The lack of a chronic Mg deficiency in rats may indicate a different mechanism of action than observed in humans. PMID- 9744850 TI - Oral taurine supplementation modulates ethanol-conditioned stimulus preference. AB - The present study investigated the possible modulatory action of oral taurine supplementation on the rewarding and aversive properties of low and high ethanol doses in male Wistar rats. A vinegar odor stimulus was daily paired with either ethanol (0.3 or 2.0 g/kg) or saline. In addition, half of the rats were supplemented orally with taurine (0.5 g/kg/day). After eight conditioning sessions, all rats were tested for their vinegar stimulus preference or aversion. In nontaurine-treated rats, 2.0 g/kg ethanol conditioning induced a significant aversion for the vinegar stimulus, while there was no preference after 0.3 g/kg ethanol conditioning. However, in taurine-supplemented rats, the 2.0 g/kg ethanol induced aversion for the stimulus was decreased significantly, while the rats administered the lower ethanol doses, 0.3 g/kg, in combination with taurine supplementation, demonstrated a significant stimulus preference. Such results suggest that taurine modulates some of the aversive or rewarding effects of ethanol. PMID- 9744851 TI - Host response to mycobacterial infection in the alcoholic rat: male and female dimorphism. AB - Increased susceptibility to tuberculosis occurs in the alcoholic. One explanation for the altered susceptibility is a change in T-lymphocyte modulation. To evaluate this, 24 male and 24 female Sprague-Dawley rats were treated with either a Lieber-type liquid ethanol diet (LED) or an isocaloric control (LCD). After 2 weeks, half the subjects were infected with BCG (10(8) colony-forming units) and sacrificed after 42 days. Splenic helper (CD4) and suppressor/cytoxic (CD8) cells were quantitated by flow cytometry. By three-way analysis of variance, splenic cellularity was significantly increased by infection (p < 0.0001) but suppressed by LED (p = 0.0002). There was a marginal sexual difference (p = 0.065) with females exhibiting a 35% lower response while on alcohol. Examining lymphocyte subsets, the most significant changes were observed after infection (BCG) and alcohol treatment (LED). CD4 levels were diminished by LED (p = 0.0002) but markedly increased by infection (p < 0.0001), producing a highly significant interaction that affected both absolute number (p < 0.0001) and relative percent present (p = 0.0078). CD8 was influenced only by infection (p < 0.0001). This resulted in a infection-related increase in the CD4/CD8 ratio which was lower with LED (p = 0.0032). Splenic T-lymphocytes, predominately CD4, are involved in the host response to BCG hepatitis and are adversely influenced by LED, which may contribute to increased susceptibility. PMID- 9744852 TI - Time-dependent effects of acute ethanol administration on regional cerebral blood flow in the rat. AB - The present study investigated the role of the postinjection interval in determining the functional consequences of acute ethanol administration in the CNS. Regional cerebral blood flow (RCBF) was determined by the [14C]iodoantipyrine method in 33 brain structures of ethanol-naive Sprague-Dawley rats. In the first experiment, changes in RCBF were assessed 5 and 15 min after a 0.8 g/kg (i.p.) dose of ethanol or water. Five minutes after treatment, rates of RCBF were increased in the motor cortex, agranular insular cortex, and the olfactory tubercle compared to water controls. No significant differences compared to control were found at the 15-min time point, despite the continued presence of ethanol in the blood. Experiment 2 tested whether blood ethanol level was the sole determinant of this response to ethanol by comparing animals with the same blood ethanol level at the 5- and 15-min time points. Greater rates of RCBF were found at 5 min postinjection compared to 15 min, in the motor cortex, agranular insular cortex, caudate/putamen, cerebellum, and the lateral septum. These data demonstrate that the rates of cerebral blood flow are increased in regionally discrete portions of the rat brain shortly after ethanol administration. Furthermore, blood ethanol level is not the exclusive factor governing this functional response. PMID- 9744853 TI - Differential effects of methanol on rat aortic smooth muscle. AB - The effects of methanol on isolated segments of rat aorta were investigated. In the absence of any vasoactive agent, methanol (5-675 mM) failed to alter basal tension. In rat aortic rings precontracted with high K+ (30 mM), methanol elicited a concentration-related relaxation at concentrations of from 5 to 675 mM. The K+-induced contraction in the presence of endothelium was more strongly inhibited by methanol than in the absence of endothelium. The effective concentration producing approximately 50% of the maximal relaxation response (ED50) to methanol was about 96 mM. Methanol-induced relaxations could not be abolished either by 5 x 10(-5) M N-nitro-L-arginine methyl ester (L-NAME) or NG nitro-L-arginine (L-NNA), both selective inhibitors of nitric oxide (NO) formation; these relaxations were not potentiated by addition of excess L arginine. An inhibitor of prostanoid synthesis, indomethacin (10(-5) M), had no effects on methanol-induced relaxation. Removal of extracellular Ca2+ ([Ca2+]o) resulted in almost complete inhibition of the relaxant effects of methanol on rat aortic ring segments. Marked attenuation of the relaxation responses of intact arteries to methanol was obtained after buffering intracellular Ca2+ ([Ca2+]i) with 10 microM BAPTA-AM. In 5-hydroxytryptamine (5-HT, 2.5 microM)- or phenylephrine (PE, 0.1 microM)-precontracted rat aortic rings, methanol amplified contractile responses to 5-HT and PE; these increased responses were concentration dependent. No significant differences in these methanol potentiated responses were found between aorta with or without endothelial cells. The amplified rat aortic smooth muscle responses induced by methanol after PE could be modified only by phentolamine, an antagonist of PE, while responses to 5-HT could be inhibited by methysergide (an antagonist of 5-HT) and by phentolamine, diphenhydramine, and haloperidol. Pretreatment with 50, 200, and 500 mM methanol increased rat aortic contractile responses induced by 5-HT and PE. Our results suggest that: (a) acute methanol exposure relaxes rat aortic smooth muscle contractile responses induced by high K+, leading to vessel relaxation. This relaxation effect of methanol is endothelium-dependent, clearly Ca2+ dependent, and independent of endogenous vasodilators such as acetylcholine, histamine, catecholamines, serotonin, or PG. (b) Methanol seems to increase potassium current by shifting the potential towards more negative values in depolarized vascular muscle cell membranes, probably inducing hyperpolarization of the cell membranes leading to a repolarization. (c) In contrast to the relaxant responses, methanol potentiates contractile response of rat aorta to 5-HT and PE. PMID- 9744854 TI - The drinking day as a unit of exposure in the epidemiology of alcohol-related medical disorders. AB - Tissue or organ system damage resulting from alcohol ingestion typically requires several years of heavy drinking to reach clinical significance. Based upon earlier empirical findings and theorizing, we hypothesized that the lifetime number of exposures (drinking days) may be of significance in understanding the relationship between chronic alcohol consumption and organ system perturbations in alcoholic populations. To test this hypothesis, detailed lifetime alcohol consumption histories from a racially mixed cohort of detoxified alcoholics (n = 253) and nonalcoholics (n = 61) were examined to determine the lifetime total number of drinking days. Linear regressions corrected for lifetime total dose and pertinent confounding variables yielded statistically significant correlations of moderate size of the number of lifetime alcohol drinking days with diastolic blood pressure and quadriceps muscle strength. The findings were considered to provide evidence that an alcohol exposure (drinking day), independent of dose, is a biologically significant event in the genesis of tissue toxicities in the cohorts studied. PMID- 9744855 TI - Neonatal alcohol exposure produces hyperactivity in high-alcohol-sensitive but not in low-alcohol-sensitive rats. AB - Children of women who consume high amounts of alcohol during their pregnancies vary greatly in physical and behavioral outcomes. Although many factors, such as dose and timing of exposure, undoubtedly contribute to this variation, one important determinant may be genetic differences in the response to alcohol. The present study examined activity levels in high alcohol sensitivity (HAS) and low alcohol sensitivity (LAS) rats following neonatal alcohol exposure. These lines were selectively bred for extremes in ethanol-induced "sleep times." The HAS and LAS offspring were exposed to alcohol via an artificial rearing procedure using the "pup-in-the-cup" technique. Rat pups were exposed to ethanol (6 g/kg/day) from postnatal day (PD) 4 through 7 and faded to a dose of 3 g/kg/day on PD 8 and 9. An artificially reared gastrostomy control group (GC) and a normally reared suckle control group (SC) were also included. Activity level was measured on PD 18 through PD 21 for 30 min daily in automated activity monitors. Neonatal ethanol exposure produced overactivity in HAS rats, relative to their controls, but the same ethanol treatment had no effect on the LAS rats. Importantly, there were no differences in blood alcohol concentrations (around 420 mg/dl) between the two lines during the treatment period. These data suggest that genetic differences in response to alcohol may be a predictor for some of the behavioral teratogenic effects of alcohol. PMID- 9744856 TI - Lifelong ethanol consumption and loss of locus coeruleus neurons in AA and ANA rats. AB - The effects of lifelong ethanol exposure and aging on the morphology of the locus coeruleus (LC) were studied in the AA (Alko, Alcohol) and ANA (Alko, Nonalcohol) rats of both sexes. The ethanol-consuming (EtOH) rats were given 12% (v/v) ethanol as the only drinking fluid from 4 to 22 months of age, whereas the young (3-month-old) and aged (24-month-old) controls had only water available. The total LC neuron numbers were obtained by using the unbiased disector method. In the AA line, as we have previously reported. the EtOH female and male rats displayed a 26-30% loss of LC neurons compared with the controls. In the ANA line, the EtOH females had 30% fewer LC neurons than the controls (EtOH 1579 +/- 377 vs. controls 2264 +/- 269, ANOVA p < 0.01), whereas the EtOH males showed no neuron loss compared to the controls (EtOH 1848 +/- 525 vs. controls 2216 +/- 152, ANOVA NS). However, taking into account (sex by line ANCOVA) the markedly higher ethanol intake of the female rats in both lines, no gender or line differences in the ethanol-induced LC degeneration were detected. Neither was there any difference in LC neuron numbers between the young and old control rats of either line of rats. In conclusion, chronic alcohol consumption, not aging per se, damages the LC neurons in experimental animals. PMID- 9744857 TI - An investigation into the effects of 5-HT agonists and receptor antagonists on ethanol self-administration in the rat. AB - Pharmacological manipulation leading to altered 5-HT function has been widely demonstrated to reduce ethanol intake in free choice tests. The aim of the present study was to examine the effects of a range of compounds known to influence 5-HT neurotransmission, including selective 5-HT receptor agonists and antagonists, on ethanol ingestion and maintained behaviour in an operant self administration paradigm. Female Sprague-Dawley rats were trained to respond for 8% ethanol (v/v) in a 60-min test by a previously described technique. The number of responses and ethanol reinforcers (dipper deliveries), ethanol consumption (g/kg of body weight), and locomotor activity (LMA) were measured following administration of 5-HT agonists (5-HT, d-fenfluramine, fluoxetine, buspirone, TFMPP, and DOI) and antagonists (metergoline, ritanserin, and ondansetron) 30 min prior to testing. d-Fenfluramine, fluoxetine, buspirone, TFMPP, and DOI all produced a reduction in ethanol ingestion and maintained behaviour at doses that failed to reduce LMA. Conversely, metergoline and ritanserin only reduced ethanol self-administration at doses that concomitantly reduced LMA. 5-HT and ondansetron were without effect on any measure. These results demonstrate that, under the present experimental conditions, activation of central 5-HT1A, 5-HT1B, and 5-HT2 receptors reduced ethanol intake and reinforced behaviour in an operant paradigm. PMID- 9744858 TI - Control of MAP kinase signaling specificity or how not to go HOG wild. PMID- 9744859 TI - Tumor necrosis factor receptor-associated factors (TRAFs)--a family of adapter proteins that regulates life and death. PMID- 9744860 TI - DNA damage activates p53 through a phosphorylation-acetylation cascade. AB - Activation of p53-mediated transcription is a critical cellular response to DNA damage. p53 stability and site-specific DNA-binding activity and, therefore, transcriptional activity, are modulated by post-translational modifications including phosphorylation and acetylation. Here we show that p53 is acetylated in vitro at separate sites by two different histone acetyltransferases (HATs), the coactivators p300 and PCAF. p300 acetylates Lys-382 in the carboxy-terminal region of p53, whereas PCAF acetylates Lys-320 in the nuclear localization signal. Acetylations at either site enhance sequence-specific DNA binding. Using a polyclonal antisera specific for p53 that is phosphorylated or acetylated at specific residues, we show that Lys-382 of human p53 becomes acetylated and Ser 33 and Ser-37 become phosphorylated in vivo after exposing cells to UV light or ionizing radiation. In vitro, amino-terminal p53 peptides phosphorylated at Ser 33 and/or at Ser-37 differentially inhibited p53 acetylation by each HAT. These results suggest that DNA damage enhances p53 activity as a transcription factor in part through carboxy-terminal acetylation that, in turn, is directed by amino terminal phosphorylation. PMID- 9744861 TI - Mitotic inactivation of a human SWI/SNF chromatin remodeling complex. AB - During mitosis, chromatin is condensed into mitotic chromosomes and transcription is inhibited, processes that might be opposed by the chromatin remodeling activity of the SWI/SNF complexes. Brg1 and hBrm, which are components of human SWI/SNF (hSWI/SNF) complexes, were recently shown to be phosphorylated during mitosis. This suggested that phosphorylation might be used as a switch to modulate SWI/SNF activity. Using an epitope-tag strategy, we have purified hSWI/SNF complexes at different stages of the cell cycle, and found that hSWI/SNF was inactive in cells blocked in G2-M. Mitotic hSWI/SNF contained Brg1 but not hBrm, and was phosphorylated on at least two subunits, hSWI3 and Brg1. In vitro, active hSWI/SNF from asynchronous cells can be phosphorylated and inactivated by ERK1, and reactivated by dephosphorylation. hSWI/SNF isolated as cells traversed mitosis regained activity when its subunits were dephosphorylated either in vitro or in vivo. We propose that this transitional inactivation and reactivation of hSWI/SNF is required for formation of a repressed chromatin structure during mitosis and reformation of an active chromatin structure as cells leave mitosis. PMID- 9744862 TI - Loss of transcriptional activity of a transgene is accompanied by DNA methylation and histone deacetylation and is prevented by insulators. AB - The constitutive DNase I hypersensitive site at the 5' end of the chicken beta globin locus marks the boundary of the active chromatin domain in erythroid cells. The DNA sequence containing this site has the properties of an insulator, as shown by its ability in stable transformation experiments to block enhancer promoter interaction when it lies between the two, but not when it lies outside, and to protect against position effects in Drosophila. We now show that the chicken insulator can protect a stably integrated gene, which is otherwise subject to great variability of expression, from chromatin-mediated repression in cell culture. When the integrated reporter gene is surrounded by insulator elements, stably transformed cell lines display consistent enhancer-dependent expression levels, in accord with the strength of the enhancer. In the absence of insulators, long-term nonselective propagation of cells carrying the integrated reporter gene results in gradual extinction of the reporter's expression, with expression patterns from tandemly repeated inserted genes suggesting that the extinction of adjacent genes is coupled. We show that the uninsulated reporter genes, in addition to becoming transcriptionally inactive, lose several epigenetic hallmarks of active chromatin, including nuclease accessibility, DNA hypomethylation, and histone hyperacetylation during time in culture. Treatment with inhibitors of histone deacetylase or DNA methylation reverses the extinction of the uninsulated genes. Extinction is completely prevented by flanking the reporter construct with insulators. Furthermore, in contrast to the uninsulated reporter genes, chromatin over the insulated genes retains nuclease accessibility and histone hyperacetylation. However, there is no clear correlation between the presence of the insulators and the level of DNA methylation. This leads us to propose a model for the insulator's ability to protect against extinction in the transformed cell lines and to function as a chromatin boundary for the chicken beta-globin locus in normal erythroid cells. PMID- 9744863 TI - Altered DNA-binding specificity mutants of EKLF and Sp1 show that EKLF is an activator of the beta-globin locus control region in vivo. AB - The locus control region of the beta-globin cluster contains five DNase I hypersensitive sites (5'HS1-5) required for locus activation. 5'HS3 contains six G-rich motifs that are essential for its activity. Members of a protein family, characterized by three zinc fingers highly homologous to those found in transcription factor Sp1, interact with these motifs. Because point mutagenesis cannot distinguish between family members, it is not known which protein activates 5'HS3. We show that the function of such closely related proteins can be distinguished in vivo by matching point mutations in 5'HS3 with amino acid changes in the zinc fingers of Sp1 and EKLF. Testing their activity in transgenic mice shows that EKLF is a direct activator of 5'HS3. PMID- 9744864 TI - The Hog1 MAPK prevents cross talk between the HOG and pheromone response MAPK pathways in Saccharomyces cerevisiae. AB - The MAPKKK Ste11p functions in three Saccharomyces cerevisiae MAPK cascades [the high osmolarity glycerol (HOG), pheromone response, and pseudohyphal/invasive growth pathways], but its activation in response to high osmolarity stimulates only the HOG pathway. To determine what restricts cross-activation of MAPK cascades (cross talk), we have studied mutants in which the pheromone response pathway is activated by high osmolarity (1 M sorbitol). We found that mutations in the HOG1 gene, encoding the p38-type MAPK of the HOG pathway, and in the PBS2 gene, encoding the activating kinase for Hog1p, allowed osmolarity-induced activation of the pheromone response pathway. This cross talk required the osmosensor Sho1p, as well as Ste20p, Ste50p, the pheromone response MAPK cascade (Ste11p, Ste7p, and Fus3p or Kss1p), and Ste12p but not Ste4p or the MAPK scaffold protein, Ste5p. The cross talk in hog1 mutants induced multiple responses of the pheromone response pathway: induction of a FUS1::lacZ reporter, morphological changes, and mating in ste4 and ste5 mutants. We suggest that Hog1p may prevent osmolarity-induced cross talk by inhibiting Sho1p, perhaps as part of a feedback control on the HOG pathway. We have also shown that Ste20p and Ste50p function in the Sho1p branch of the HOG pathway and that a second osmosensor in addition to Sho1p may activate Ste11p. Finally, we have found that pseudohyphal growth exhibited by wild-type (HOG1) strains depends on SHO1, suggesting that Sho1p may be a receptor that feeds into the pseudohyphal growth pathway. PMID- 9744865 TI - Repression of yeast Ste12 transcription factor by direct binding of unphosphorylated Kss1 MAPK and its regulation by the Ste7 MEK. AB - The mitogen-activated protein kinase (MAPK) Kss1 has a dual role in regulating filamentous (invasive) growth of the yeast Saccharomyces cerevisiae. The stimulatory function of Kss1 requires both its catalytic activity and its activation by the MAPK/ERK kinase (MEK) Ste7; in contrast, the inhibitory function of Kss1 requires neither. This study examines the mechanism by which Kss1 inhibits invasive growth, and how Ste7 action overcomes this inhibition. We found that unphosphorylated Kss1 binds directly to the transcription factor Ste12, that this binding is necessary for Kss1-mediated repression of Ste12, and that Ste7-mediated phosphorylation of Kss1 weakens Kss1-Ste12 interaction and relieves Kss1-mediated repression. Relative to Kss1, the MAPK Fus3 binds less strongly to Ste12 and is correspondingly a weaker inhibitor of invasive growth. Analysis of Kss1 mutants indicated that the activation loop of Kss1 controls binding to Ste12. Potent repression of a transcription factor by its physical interaction with the unactivated isoform of a protein kinase, and relief of this repression by activation of the kinase, is a novel mechanism for signal-dependent regulation of gene expression. PMID- 9744866 TI - CDK inhibitors p18(INK4c) and p27(Kip1) mediate two separate pathways to collaboratively suppress pituitary tumorigenesis. AB - INK4 and CIP/KIP are two distinct families of cyclin-dependent kinase (CDK) inhibitors implicated in mediating a wide range of cell growth control signals. We have created p18(INK4c)-deficient mice. These mice develop gigantism and widespread organomegaly. The pituitary gland, spleen, and thymus are disproportionately enlarged and hyperplastic. T and B lymphocytes develop normally in p18-deficient mice, but both exhibit increased cellularity and a higher proliferative rate upon mitogenic stimulation. Loss of p18, like that of p27, but not other CDK inhibitor genes, leads to a gradual progression from intermediate lobe pituitary hyperplasia in young mice to an adenoma by 10 months of age with a nearly complete penetrance. Mice lacking both p18 and p27, like mice chimeric for Rb deficiency, invariably died from pituitary adenomas by 3 months. Hence, p18 and p27 mediate two separate pathways to collaboratively suppress pituitary tumorigenesis, likely by controlling the function of Rb. PMID- 9744867 TI - Beadex encodes an LMO protein that regulates Apterous LIM-homeodomain activity in Drosophila wing development: a model for LMO oncogene function. AB - Formation of the dorsal-ventral axis of the Drosophila wing depends on activity of the LIM-homeodomain protein Apterous (Ap). Here we report that Ap activity levels are modulated by dLMO, the protein encoded by the Beadex (Bx) gene. Overexpression of dLMO in Bx mutants interferes with Apterous function. Conversely, Bx loss-of-function mutants fail to down-regulate Apterous activity at late stages of wing development. Biochemical analysis shows that dLMO protein competes for binding of Apterous to its cofactor Chip. These data suggest that Apterous activity depends on formation of a functional complex with Chip and that the relative levels of dLMO, Apterous, and Chip determine the level of Apterous activity. The dominant interference mechanism of dLMO action may serve as a model for the mechanism by which LMO oncogenes cause cancer when misexpressed in T cells. PMID- 9744868 TI - Developmentally programmed assembly of higher order telomerase complexes with distinct biochemical and structural properties. AB - In Euplotes crassus, telomerase is responsible for telomere maintenance during vegetative growth and de novo telomere synthesis during macronuclear development. Here we show that telomerase in the vegetative stage of the life cycle exists as a 280-kD complex that can add telomeric repeats only onto telomeric DNA primers. Following the initiation of macronuclear development, telomerase assembles into larger complexes of 550 kD, 1600 kD, and 5 MD. In the 1600-kDa and 5-MDa complexes, telomerase is more processive than in the two smaller complexes and can add telomeres de novo onto nontelomeric 3' ends. Assembly of higher order telomerase complexes is accompanied by an extended region of RNase V1 and RNase T1 protection in the telomerase RNA subunit that is not observed with telomerase from vegetatively growing cells. The protected residues encompass a highly conserved region previously proposed to serve as a platform for formation of higher order structures. These findings provide the first direct demonstration of developmentally regulated higher order telomerase complexes with unique biochemical and structural properties. PMID- 9744869 TI - mei-W68 in Drosophila melanogaster encodes a Spo11 homolog: evidence that the mechanism for initiating meiotic recombination is conserved. AB - Meiotic recombination requires the action of several gene products in both Saccharomyces cerevisiae and Drosophila melanogaster. Genetic studies in D. melanogaster have shown that the mei-W68 gene is required for all meiotic gene conversion and crossing-over. We cloned mei-W68 using a new genetic mapping method in which P elements are used to promote crossing-over at their insertion sites. This resulted in the high-resolution mapping of mei-W68 to a <18-kb region that contains a homolog of the S. cerevisiae spo11 gene. Molecular analysis of several mutants confirmed that mei-W68 encodes an spo11 homolog. Spo11 and MEI W68 are members of a family of proteins similar to a novel type II topoisomerase. On the basis of this and other lines of evidence, Spo11 has been proposed to be the enzymatic activity that creates the double-strand breaks needed to initiate meiotic recombination. This raises the possibility that recombination in Drosophila is also initiated by double-strand breaks. Although these homologous genes are required absolutely for recombination in both species, their roles differ in other respects. In contrast to spo11, mei-W68 is not required for synaptonemal complex formation and does have a mitotic role. PMID- 9744870 TI - Saccharomyces cerevisiae cAMP-dependent protein kinase controls entry into stationary phase through the Rim15p protein kinase. AB - The Saccharomyces cerevisiae protein kinase Rim15p was identified previously as a stimulator of meiotic gene expression. Here, we show that loss of Rim15p causes an additional pleiotropic phenotype in cells grown to stationary phase on rich medium; this phenotype includes defects in trehalose and glycogen accumulation, in transcriptional derepression of HSP12, HSP26, and SSA3, in induction of thermotolerance and starvation resistance, and in proper G1 arrest. These phenotypes are commonly associated with hyperactivity of the Ras/cAMP pathway. Tests of epistasis suggest that Rim15p may act in this pathway downstream of the cAMP-dependent protein kinase (cAPK). Accordingly, deletion of RIM15 suppresses the growth defect of a temperature-sensitive adenylate-cyclase mutant and, most importantly, renders cells independent of cAPK activity. Conversely, overexpression of RIM15 suppresses phenotypes associated with a mutation in the regulatory subunit of cAPK, exacerbates the growth defect of strains compromised for cAPK activity, and partially induces a starvation response in logarithmically growing wild-type cells. Biochemical analyses reveal that cAPK-mediated in vitro phosphorylation of Rim15p strongly inhibits its kinase activity. Taken together, these results place Rim15p immediately downstream and under negative control of cAPK and define a positive regulatory role of Rim15p for entry into both meiosis and stationary phase. PMID- 9744871 TI - Recovery from DNA replicational stress is the essential function of the S-phase checkpoint pathway. AB - RAD53 and MEC1 are essential genes required for the transcriptional and cell cycle responses to DNA damage and DNA replication blocks. We have examined the essential function of these genes and found that their lethality but not their checkpoint defects can be suppressed by increased expression of genes encoding ribonucleotide reductase. Analysis of viable null alleles revealed that Mec1 plays a greater role in response to inhibition of DNA synthesis than Rad53. The loss of survival in mec1 and rad53 null or point mutants in response to transient inhibition of DNA synthesis is not a result of inappropriate anaphase entry but primarily to an inability to complete chromosome replication. We propose that this checkpoint pathway plays an important role in the maintenance of DNA synthetic capabilities when DNA replication is stressed. PMID- 9744872 TI - The heterogeneity of ER Ca2+ stores has a key role in nonmuscle cell signaling and function. PMID- 9744873 TI - Regulation of endosome sorting by a specific PP2A isoform. AB - The regulated sorting of proteins within the trans-Golgi network (TGN)/endosomal system is a key determinant of their biological activity in vivo. For example, the endoprotease furin activates of a wide range of proproteins in multiple compartments within the TGN/endosomal system. Phosphorylation of its cytosolic domain by casein kinase II (CKII) promotes the localization of furin to the TGN and early endosomes whereas dephosphorylation is required for efficient transport between these compartments (Jones, B.G., L. Thomas, S.S. Molloy, C.D. Thulin, M.D. Fry, K.A. Walsh, and G. Thomas. 1995. EMBO [Eur. Mol. Biol. Organ.] J. 14:5869-5883). Here we show that phosphorylated furin molecules internalized from the cell surface are retained in a local cycling loop between early endosomes and the plasma membrane. This cycling loop requires the phosphorylation state dependent furin-sorting protein PACS-1, and mirrors the trafficking pathway described recently for the TGN localization of furin (Wan, L., S.S. Molloy, L. Thomas, G. Liu, Y. Xiang, S.L. Ryback, and G. Thomas. 1998. Cell. 94:205-216). We also demonstrate a novel role for protein phosphatase 2A (PP2A) in regulating protein localization in the TGN/endosomal system. Using baculovirus recombinants expressing individual PP2A subunits, we show that the dephosphorylation of furin in vitro requires heterotrimeric phosphatase containing B family regulatory subunits. The importance of this PP2A isoform in directing the routing of furin from early endosomes to the TGN was established using SV-40 small t antigen as a diagnostic tool in vivo. The role of both CKII and PP2A in controlling multiple sorting steps in the TGN/endosomal system indicates that the distribution of itinerant membrane proteins may be acutely regulated via signal transduction pathways. PMID- 9744874 TI - Annexin XIIIb associates with lipid microdomains to function in apical delivery. AB - A member of the annexin XIII sub-family, annexin XIIIb, has been implicated in the apical exocytosis of epithelial kidney cells. Annexins are phospholipid binding proteins that have been suggested to be involved in membrane trafficking events although their actual physiological function remains open. Unlike the other annexins, annexin XIIIs are myristoylated. Here, we show by immunoelectron microscopy that annexin XIIIb is localized to the trans-Golgi network (TGN), vesicular carriers and the apical cell surface. Polarized apical sorting involves clustering of apical proteins into dynamic sphingolipid-cholesterol rafts. We now provide evidence for the raft association of annexin XIIIb. Using in vitro assays and either myristoylated or unmyristoylated recombinant annexin XIIIb, we demonstrate that annexin XIIIb in its native myristoylated form stimulates specifically apical transport whereas the unmyristoylated form inhibits this route. Moreover, we show that formation of apical carriers from the TGN is inhibited by an anti-annexin XIIIb antibody whereas it is stimulated by myristoylated recombinant annexin XIIIb. These results suggest that annexin XIIIb directly participates in apical delivery. PMID- 9744876 TI - The muscle regulatory factors MyoD and myf-5 undergo distinct cell cycle-specific expression in muscle cells. AB - The muscle regulators MyoD and Myf-5 control cell cycle withdrawal and induction of differentiation in skeletal muscle cells. By immunofluorescence analysis, we show that MyoD and Myf-5 expression patterns become mutually exclusive when C2 cells are induced to differentiate with Myf-5 staining present in cells which fail to differentiate. Isolation of these undifferentiated cells reveals that upon serum stimulation they reenter the cell cycle, express MyoD and downregulate Myf-5. Similar regulations of MyoD and Myf-5 were observed using cultured primary myoblasts derived from satellite cells. To further analyze these regulations of MyoD and Myf-5 expression, we synchronized proliferating myoblasts. Analysis of MyoD and Myf-5 expression during cell cycle progression revealed distinct and contrasting profiles of expression. MyoD is absent in G0, peaks in mid-G1, falls to its minimum level at G1/S and reaugments from S to M. In contrast, Myf-5 protein is high in G0, decreases during G1 and reappears at the end of G1 to remain stable until mitosis. These data demonstrate that the two myogenic factors MyoD and Myf-5 undergo specific and distinct cell cycle-dependent regulation, thus establishing a correlation between the cell cycle-specific ratios of MyoD and Myf-5 and the capacity of cells to differentiate: (a) in G1, when cells express high levels of MyoD and enter differentiation; (b) in G0, when cells express high levels of Myf-5 and fail to differentiate. PMID- 9744875 TI - Analysis of GLUT4 distribution in whole skeletal muscle fibers: identification of distinct storage compartments that are recruited by insulin and muscle contractions. AB - The effects of insulin stimulation and muscle contractions on the subcellular distribution of GLUT4 in skeletal muscle have been studied on a preparation of single whole fibers from the rat soleus. The fibers were labeled for GLUT4 by a preembedding technique and observed as whole mounts by immunofluorescence microscopy, or after sectioning, by immunogold electron microscopy. The advantage of this preparation for cells of the size of muscle fibers is that it provides global views of the staining from one end of a fiber to the other and from one side to the other through the core of the fiber. In addition, the labeling efficiency is much higher than can be obtained with ultracryosections. In nonstimulated fibers, GLUT4 is excluded from the plasma membrane and T tubules. It is distributed throughout the muscle fibers with approximately 23% associated with large structures including multivesicular endosomes located in the TGN region, and 77% with small tubulovesicular structures. The two stimuli cause translocation of GLUT4 to both plasma membrane and T tubules. Quantitation of the immunogold electron microscopy shows that the effects of insulin and contraction are additive and that each stimulus recruits GLUT4 from both large and small depots. Immunofluorescence double labeling for GLUT4 and transferrin receptor (TfR) shows that the small depots can be further subdivided into TfR-positive and TfR-negative elements. Interestingly, we observe that colocalization of TfR and GLUT4 is increased by insulin and decreased by contractions. These results, supported by subcellular fractionation experiments, suggest that TfR-positive depots are only recruited by contractions. We do not find evidence for stimulation-induced unmasking of resident surface membrane GLUT4 transporters or for dilation of the T tubule system (Wang, W., P.A. Hansen, B.A. Marshall, J.O. Holloszy, and M. Mueckler. 1996. J. Cell Biol. 135:415-430). PMID- 9744877 TI - Progressive muscular dystrophy in alpha-sarcoglycan-deficient mice. AB - Limb-girdle muscular dystrophy type 2D (LGMD 2D) is an autosomal recessive disorder caused by mutations in the alpha-sarcoglycan gene. To determine how alpha-sarcoglycan deficiency leads to muscle fiber degeneration, we generated and analyzed alpha-sarcoglycan- deficient mice. Sgca-null mice developed progressive muscular dystrophy and, in contrast to other animal models for muscular dystrophy, showed ongoing muscle necrosis with age, a hallmark of the human disease. Sgca-null mice also revealed loss of sarcolemmal integrity, elevated serum levels of muscle enzymes, increased muscle masses, and changes in the generation of absolute force. Molecular analysis of Sgca-null mice demonstrated that the absence of alpha-sarcoglycan resulted in the complete loss of the sarcoglycan complex, sarcospan, and a disruption of alpha-dystroglycan association with membranes. In contrast, no change in the expression of epsilon sarcoglycan (alpha-sarcoglycan homologue) was observed. Recombinant alpha sarcoglycan adenovirus injection into Sgca-deficient muscles restored the sarcoglycan complex and sarcospan to the membrane. We propose that the sarcoglycan-sarcospan complex is requisite for stable association of alpha dystroglycan with the sarcolemma. The Sgca-deficient mice will be a valuable model for elucidating the pathogenesis of sarcoglycan deficient limb-girdle muscular dystrophies and for the development of therapeutic strategies for this disease. PMID- 9744878 TI - A role for a protease in morphogenic responses during yeast cell fusion. AB - Cell fusion during yeast mating provides a model for signaling-controlled changes at the cell surface. We identified the AXL1 gene in a screen for genes required for cell fusion in both mating types during mating. AXL1 is a pheromone-inducible gene required for axial bud site selection in haploid yeast and for proteolytic maturation of a-factor. Two other bud site selection genes, RSR1, encoding a small GTPase, and BUD3, were also required for efficient cell fusion. Based on double mutant analysis, AXL1 in a MATalpha strain acted genetically in the same pathway with FUS2, a fusion-dedicated gene. Electron microscopy of axl1, rsr1, and fus2 prezygotes revealed similar defects in nuclear migration, vesicle accumulation, cell wall degradation, and membrane fusion during cell fusion. The axl1 and rsr1 mutants exhibited defects in pheromone-induced morphogenesis. AXL1 protease function was required in MATalpha strains for fusion during mating. The ability of the Rsr1p GTPase to cycle was required for efficient cell fusion, as it is for bud site selection. During conjugation, vegetative functions may be redeployed under the control of pheromone signaling for mating purposes. Since Rsr1p has been reported to physically associate with Cdc24p and Bem1p components of the pheromone response pathway, we suggest that the bud site selection genes Rsr1p and Axl1p may act to mediate pheromone control of Fus2p-based fusion events during mating. PMID- 9744879 TI - A morphogenesis checkpoint monitors the actin cytoskeleton in yeast. AB - A morphogenesis checkpoint in budding yeast delays cell cycle progression in response to perturbations of cell polarity that prevent bud formation (Lew, D.J., and S.I. Reed. 1995. J. Cell Biol. 129:739- 749). The cell cycle delay depends upon the tyrosine kinase Swe1p, which phosphorylates and inhibits the cyclin dependent kinase Cdc28p (Sia, R.A.L., H.A. Herald, and D.J. Lew. 1996. Mol. Biol. Cell. 7:1657- 1666). In this report, we have investigated the nature of the defect(s) that trigger this checkpoint. A Swe1p- dependent cell cycle delay was triggered by direct perturbations of the actin cytoskeleton, even when polarity establishment functions remained intact. Furthermore, actin perturbation could trigger the checkpoint even in cells that had already formed a bud, suggesting that the checkpoint directly monitors actin organization, rather than (or in addition to) polarity establishment or bud formation. In addition, we show that the checkpoint could detect actin perturbations through most of the cell cycle. However, the ability to respond to such perturbations by delaying cell cycle progression was restricted to a narrow window of the cell cycle, delimited by the periodic accumulation of the checkpoint effector, Swe1p. PMID- 9744880 TI - Assembly and function of the actin cytoskeleton of yeast: relationships between cables and patches. AB - Actin in eukaryotic cells is found in different pools, with filaments being organized into a variety of supramolecular assemblies. To investigate the assembly and functional relationships between different parts of the actin cytoskeleton in one cell, we studied the morphology and dynamics of cables and patches in yeast. The fine structure of actin cables and the manner in which cables disassemble support a model in which cables are composed of a number of overlapping actin filaments. No evidence for intrinsic polarity of cables was found. To investigate to what extent different parts of the actin cytoskeleton depend on each other, we looked for relationships between cables and patches. Patches and cables were often associated, and their polarized distributions were highly correlated. Therefore, patches and cables do appear to depend on each other for assembly and function. Many cell types show rearrangements of the actin cytoskeleton, which can occur via assembly or movement of actin filaments. In our studies, dramatic changes in actin polarization did not include changes in filamentous actin. In addition, the concentration of actin patches was relatively constant as cells grew. Therefore, cells do not have bursts of activity in which new parts of the actin cytoskeleton are created. PMID- 9744881 TI - Stepwise reconstitution of interphase microtubule dynamics in permeabilized cells and comparison to dynamic mechanisms in intact cells. AB - Microtubules in permeabilized cells are devoid of dynamic activity and are insensitive to depolymerizing drugs such as nocodazole. Using this model system we have established conditions for stepwise reconstitution of microtubule dynamics in permeabilized interphase cells when supplemented with various cell extracts. When permeabilized cells are supplemented with mammalian cell extracts in the presence of protein phosphatase inhibitors, microtubules become sensitive to nocodazole. Depolymerization induced by nocodazole proceeds from microtubule plus ends, whereas microtubule minus ends remain inactive. Such nocodazole sensitive microtubules do not exhibit subunit turnover. By contrast, when permeabilized cells are supplemented with Xenopus egg extracts, microtubules actively turn over. This involves continuous creation of free microtubule minus ends through microtubule fragmentation. Newly created minus ends apparently serve as sites of microtubule depolymerization, while net microtubule polymerization occurs at microtubule plus ends. We provide evidence that similar microtubule fragmentation and minus end-directed disassembly occur at the whole-cell level in intact cells. These data suggest that microtubule dynamics resembling dynamics observed in vivo can be reconstituted in permeabilized cells. This model system should provide means for in vitro assays to identify molecules important in regulating microtubule dynamics. Furthermore, our data support recent work suggesting that microtubule treadmilling is an important mechanism of microtubule turnover. PMID- 9744882 TI - Activation of the MKK/ERK pathway during somatic cell mitosis: direct interactions of active ERK with kinetochores and regulation of the mitotic 3F3/2 phosphoantigen. AB - The mitogen-activated protein (MAP) kinase pathway, which includes extracellular signal-regulated protein kinases 1 and 2 (ERK1, ERK2) and MAP kinase kinases 1 and 2 (MKK1, MKK2), is well-known to be required for cell cycle progression from G1 to S phase, but its role in somatic cell mitosis has not been clearly established. We have examined the regulation of ERK and MKK in mammalian cells during mitosis using antibodies selective for active phosphorylated forms of these enzymes. In NIH 3T3 cells, both ERK and MKK are activated within the nucleus during early prophase; they localize to spindle poles between prophase and anaphase, and to the midbody during cytokinesis. During metaphase, active ERK is localized in the chromosome periphery, in contrast to active MKK, which shows clear chromosome exclusion. Prophase activation and spindle pole localization of active ERK and MKK are also observed in PtK1 cells. Discrete localization of active ERK at kinetochores is apparent by early prophase and during prometaphase with decreased staining on chromosomes aligned at the metaphase plate. The kinetochores of chromosomes displaced from the metaphase plate, or in microtubule disrupted cells, still react strongly with the active ERK antibody. This pattern resembles that reported for the 3F3/2 monoclonal antibody, which recognizes a phosphoepitope that disappears with kinetochore attachment to the spindles, and has been implicated in the mitotic checkpoint for anaphase onset (Gorbsky and Ricketts, 1993. J. Cell Biol. 122:1311-1321). The 3F3/2 reactivity of kinetochores on isolated chromosomes decreases after dephosphorylation with protein phosphatase, and then increases after subsequent phosphorylation by purified active ERK or active MKK. These results suggest that the MAP kinase pathway has multiple functions during mitosis, helping to promote mitotic entry as well as targeting proteins that mediate mitotic progression in response to kinetochore attachment. PMID- 9744883 TI - Active MAP kinase in mitosis: localization at kinetochores and association with the motor protein CENP-E. AB - To investigate possible involvement of the mitogen-activated protein (MAP) kinases ERK1 and ERK2 (extracellular signal-regulated kinases) in somatic cell mitosis, we have used indirect immunofluorescence with a highly specific phospho MAP kinase antibody and found that a portion of the active MAP kinase is localized at kinetochores, asters, and the midbody during mitosis. Although the aster labeling was constant from the time of nuclear envelope breakdown, the kinetochore labeling first appeared at early prometaphase, started to fade during chromosome congression, and then disappeared at midanaphase. At telophase, active MAP kinase localized at the midbody. Based on colocalization and the presence of a MAP kinase consensus phosphorylation site, we identified the kinetochore motor protein CENP-E as a candidate mitotic substrate for MAP kinase. CENP-E was phosphorylated in vitro by MAP kinase on sites that are known to regulate its interactions with microtubules and was found to associate in vivo preferentially with the active MAP kinase during mitosis. Therefore, the presence of active MAP kinase at specific mitotic structures and its interaction with CENP-E suggest that MAP kinase could play a role in mitosis at least in part by altering the ability of CENP-E to mediate interactions between chromosomes and microtubules. PMID- 9744884 TI - The Xenopus Chk1 protein kinase mediates a caffeine-sensitive pathway of checkpoint control in cell-free extracts. AB - We have analyzed the role of the protein kinase Chk1 in checkpoint control by using cell-free extracts from Xenopus eggs. Recombinant Xenopus Chk1 (Xchk1) phosphorylates the mitotic inducer Cdc25 in vitro on multiple sites including Ser 287. The Xchk1-catalyzed phosphorylation of Cdc25 on Ser-287 is sufficient to confer the binding of 14-3-3 proteins. Egg extracts from which Xchk1 has been removed by immunodepletion are strongly but not totally compromised in their ability to undergo a cell cycle delay in response to the presence of unreplicated DNA. Cdc25 in Xchk1-depleted extracts remains bound to 14-3-3 due to the action of a distinct Ser-287-specific kinase in addition to Xchk1. Xchk1 is highly phosphorylated in the presence of unreplicated or damaged DNA, and this phosphorylation is abolished by caffeine, an agent which attenuates checkpoint control. The checkpoint response to unreplicated DNA in this system involves both caffeine-sensitive and caffeine-insensitive steps. Our results indicate that caffeine disrupts the checkpoint pathway containing Xchk1. PMID- 9744885 TI - Restriction of 480/270-kD ankyrin G to axon proximal segments requires multiple ankyrin G-specific domains. AB - AnkyrinG (-/-) neurons fail to concentrate voltage-sensitive sodium channels and neurofascin at their axon proximal segments, suggesting that ankyrinG is a key component of a structural pathway involved in assembly of specialized membrane domains at axon proximal segments and possibly nodes of Ranvier (Zhou, D., S. Lambert, D.L. Malen, S. Carpenter, L. Boland, and V. Bennett, manuscript submitted for publication). This paper addresses the mechanism for restriction of 270-kD ankyrinG to axon proximal segments by evaluation of localization of GFP tagged ankyrinG constructs transfected into cultured dorsal root ganglion neurons, as well as measurements of fluorescence recovery after photobleaching of neurofascin- GFP-tagged ankyrinG complexes in nonneuronal cells. A conclusion is that multiple ankyrinG-specific domains, in addition to the conserved membrane binding domain, contribute to restriction of ankyrinG to the axonal plasma membrane in dorsal root ganglion neurons. The ankyrinG-specific spectrin-binding and tail domains are capable of binding directly to sites on the plasma membrane of neuronal cell bodies and axon proximal segments, and presumably have yet to be identified docking sites. The serine-rich domain, which is present only in 480- and 270-kD ankyrinG polypeptides, contributes to restriction of ankyrinG to axon proximal segments as well as limiting lateral diffusion of ankyrinG-neurofascin complexes. The membrane-binding, spectrin-binding, and tail domains of ankyrinG also contribute to limiting the lateral mobility of ankyrinG-neurofascin complexes. AnkyrinG thus functions as an integrated mechanism involving cooperation among multiple domains heretofore regarded as modular units. This complex behavior explains ability of ankyrinB and ankyrinG to sort to distinct sites in neurons and the fact that these ankyrins do not compensate for each other in ankyrin gene knockouts in mice. PMID- 9744886 TI - Blocking cytochrome c activity within intact neurons inhibits apoptosis. AB - Cytochrome c has been shown to play a role in cell-free models of apoptosis. During NGF withdrawal-induced apoptosis of intact rat superior cervical ganglion (SCG) neurons, we observe the redistribution of cytochrome c from the mitochondria to the cytoplasm. This redistribution is not inhibited by the caspase inhibitor Z-Val-Ala-Asp-fluoromethylketone (ZVADfmk) but is blocked by either of the neuronal survival agents 8-(4-chlorophenylthio)adenosine 3':5' cyclic monophosphate (CPT-cAMP) or cycloheximide. Moreover, microinjection of SCG neurons with antibody to cytochrome c blocks NGF withdrawal-induced apoptosis. However, microinjection of SCG neurons with cytochrome c does not alter the rate of apoptosis in either the presence or absence of NGF. These data suggest that cytochrome c is an intrinsic but not limiting component of the neuronal apoptotic pathway. PMID- 9744887 TI - The symmetrical structure of structural maintenance of chromosomes (SMC) and MukB proteins: long, antiparallel coiled coils, folded at a flexible hinge. AB - Structural maintenance of chromosomes (SMC) proteins function in chromosome condensation and several other aspects of DNA processing. They are large proteins characterized by an NH2-terminal nucleotide triphosphate (NTP)-binding domain, two long segments of coiled coil separated by a hinge, and a COOH-terminal domain. Here, we have visualized by EM the SMC protein from Bacillus subtilis (BsSMC) and MukB from Escherichia coli, which we argue is a divergent SMC protein. Both BsSMC and MukB show two thin rods with globular domains at the ends emerging from the hinge. The hinge appears to be quite flexible: the arms can open up to 180 degrees, separating the terminal domains by 100 nm, or close to near 0 degrees, bringing the terminal globular domains together. A surprising observation is that the approximately 300-amino acid-long coiled coils are in an antiparallel arrangement. Known coiled coils are almost all parallel, and the longest antiparallel coiled coils known previously are 35-45 amino acids long. This antiparallel arrangement produces a symmetrical molecule with both an NH2- and a COOH-terminal domain at each end. The SMC molecule therefore has two complete and identical functional domains at the ends of the long arms. The bifunctional symmetry and a possible scissoring action at the hinge should provide unique biomechanical properties to the SMC proteins. PMID- 9744889 TI - Analog versus digital: extrapolating from electronics to neurobiology. AB - We review the pros and cons of analog and digital computation. We propose that computation that is most efficient in its use of resources is neither analog computation nor digital computation but, rather, a mixture of the two forms. For maximum efficiency, the information and information-processing resources of the hybrid form must be distributed over many wires, with an optimal signal-to-noise ratio per wire. Our results suggest that it is likely that the brain computes in a hybrid fashion and that an underappreciated and important reason for the efficiency of the human brain, which consumes only 12 W, is the hybrid and distributed nature of its architecture. PMID- 9744888 TI - The membrane-proximal region of the E-cadherin cytoplasmic domain prevents dimerization and negatively regulates adhesion activity. AB - Cadherins are transmembrane glycoproteins involved in Ca2+-dependent cell-cell adhesion. Deletion of the COOH-terminal residues of the E-cadherin cytoplasmic domain has been shown to abolish its cell adhesive activity, which has been ascribed to the failure of the deletion mutants to associate with catenins. Based on our present results, this concept needs revision. As was reported previously, leukemia cells (K562) expressing E-cadherin with COOH-terminal deletion of 37 or 71 amino acid residues showed almost no aggregation. Cells expressing E-cadherin with further deletion of 144 or 151 amino acid residues, which eliminates the membrane-proximal region of the cytoplasmic domain, showed E-cadherin-dependent aggregation. Thus, deletion of the membrane-proximal region results in activation of the nonfunctional E-cadherin polypeptides. However, these cells did not show compaction. Chemical cross-linking revealed that the activated E-cadherin polypeptides can be cross-linked to a dimer on the surface of cells, whereas the inactive polypeptides, as well as the wild-type E-cadherin polypeptide containing the membrane-proximal region, can not. Therefore, the membrane-proximal region participates in regulation of the adhesive activity by preventing lateral dimerization of the extracellular domain. PMID- 9744890 TI - Employing the zeta-transform to optimize the calculation of the synaptic conductance of NMDA and other synaptic channels in network simulations. AB - Calculation of the total conductance change induced by multiple synapses at a given membrane compartment remains one of the most time-consuming processes in biophysically realistic neural network simulations. Here we show that this calculation can be achieved in a highly efficient way even for multiply converging synapses with different delays by means of the zeta-transform. Using the example of an NMDA synapse, we show that every update of the total conductance is achieved by an iterative process requiring at most three recent multiplications, which together need only the history values from the two most recent iterations. A major advantage is that this small computational load is independent of the number of synapses simulated. A benchmark comparison to other techniques demonstrates superior performance of the zeta-transform. Nonvoltage dependent synaptic channels can be treated similarly (Olshausen, 1990; Brettle & Niebur, 1994), and the technique can also be generalized to other synaptic channels. PMID- 9744891 TI - Site-selective autophosphorylation of Ca2+/calmodulin-dependent protein kinase II as a synaptic encoding mechanism. AB - A detailed kinetic model of the Ca2+/calmodulin-dependent protein kinase II (CaMKII) is presented in which subunits undergo autophosphorylation at several sites in a manner that depends on the frequency and duration of Ca2+ spikes. It is shown that high-frequency stimulation causes autophosphorylation of the autonomy site (Thr286), and promotes persistent catalytic activity. On the other hand, low-frequency stimulation is shown to cause autophosphorylation of an inhibitory site (Thr306), which prevents subunit activation. This site-selective autophosphorylation provides the basis for a molecular switch. When activated by a strong stimulus, the switch remains on for many minutes, even in the presence of a CaMKII-specific phosphatase. However, prolonged low-frequency stimulation disables the switch, and influences the response to subsequent stimulation. It is conceivable that a regulatory mechanism such as this may permit CaMKII to mediate synaptic frequency encoding and thereby direct an appropriate change in synaptic efficacy. It is indicated how the behavior of the model may relate to the induction of long-term potentiation. PMID- 9744892 TI - Ion channel stochasticity may be critical in determining the reliability and precision of spike timing. AB - The firing reliability and precision of an isopotential membrane patch consisting of a realistically large number of ion channels is investigated using a stochastic Hodgkin-Huxley (HH) model. In sharp contrast to the deterministic HH model, the biophysically inspired stochastic model reproduces qualitatively the different reliability and precision characteristics of spike firing in response to DC and fluctuating current input in neocortical neurons, as reported by Mainen & Sejnowski (1995). For DC inputs, spike timing is highly unreliable; the reliability and precision are significantly increased for fluctuating current input. This behavior is critically determined by the relatively small number of excitable channels that are opened near threshold for spike firing rather than by the total number of channels that exist in the membrane patch. Channel fluctuations, together with the inherent bistability in the HH equations, give rise to three additional experimentally observed phenomena: subthreshold oscillations in the membrane voltage for DC input, "spontaneous" spikes for subthreshold inputs, and "missing" spikes for suprathreshold inputs. We suggest that the noise inherent in the operation of ion channels enables neurons to act as "smart" encoders. Slowly varying, uncorrelated inputs are coded with low reliability and accuracy and, hence, the information about such inputs is encoded almost exclusively by the spike rate. On the other hand, correlated presynaptic activity produces sharp fluctuations in the input to the postsynaptic cell, which are then encoded with high reliability and accuracy. In this case, information about the input exists in the exact timing of the spikes. We conclude that channel stochasticity should be considered in realistic models of neurons. PMID- 9744893 TI - Fast temporal encoding and decoding with spiking neurons. AB - We propose a simple theoretical structure of interacting integrate-and-fire neurons that can handle fast information processing and may account for the fact that only a few neuronal spikes suffice to transmit information in the brain. Using integrate-and-fire neurons that are subjected to individual noise and to a common external input, we calculate their first passage time (FPT), or interspike interval. We suggest using a population average for evaluating the FPT that represents the desired information. Instantaneous lateral excitation among these neurons helps the analysis. By employing a second layer of neurons with variable connections to the first layer, we represent the strength of the input by the number of output neurons that fire, thus decoding the temporal information. Such a model can easily lead to a logarithmic relation as in Weber's law. The latter follows naturally from information maximization if the input strength is statistically distributed according to an approximate inverse law. PMID- 9744895 TI - Mutual information, Fisher information, and population coding. AB - In the context of parameter estimation and model selection, it is only quite recently that a direct link between the Fisher information and information theoretic quantities has been exhibited. We give an interpretation of this link within the standard framework of information theory. We show that in the context of population coding, the mutual information between the activity of a large array of neurons and a stimulus to which the neurons are tuned is naturally related to the Fisher information. In the light of this result, we consider the optimization of the tuning curves parameters in the case of neurons responding to a stimulus represented by an angular variable. PMID- 9744894 TI - Linearization of F-I curves by adaptation. AB - We show that negative feedback to highly nonlinear frequency-current (F-I) curves results in an effective linearization. (By highly nonlinear we mean that the slope at threshold is infinite or very steep.) We then apply this to a specific model for spiking neurons and show that the details of the adaptation mechanism do not affect the results. The crucial points are that the adaptation is slow compared to other processes and the unadapted F-I curve is highly nonlinear. PMID- 9744896 TI - Synaptic pruning in development: a computational account. AB - Research with humans and primates shows that the developmental course of the brain involves synaptic overgrowth followed by marked selective pruning. Previous explanations have suggested that this intriguing, seemingly wasteful phenomenon is utilized to remove, "erroneous" synapses. We prove that this interpretation is wrong if synapses are Hebbian. Under limited metabolic energy resources restricting the amount and strength of synapses, we show that memory performance is maximized if synapses are first overgrown and then pruned following optimal "minimal-value" deletion. This optimal strategy leads to interesting insights concerning childhood amnesia. PMID- 9744897 TI - Spatial decorrelation in orientation-selective cortical cells. AB - We propose a model for the lateral connectivity of orientation-selective cells in the visual cortex. We study the properties of the input signal to the visual cortex and find new statistical structures that have not been processed in the retino-geniculate pathway. Using the idea that the system performs redundancy reduction of the incoming signals, we derive the lateral connectivity that will achieve this for a set of orientation-selective local circuits, as well as the complete spatial structure of a network composed of such circuits. We compare the results with various physiological measurements. PMID- 9744898 TI - Receptive Field Formation in Natural Scene Environments. Comparison of Single Cell Learning Rules. AB - We study several statistically and biologically motivated learning rules using the same visual environment: one made up of natural scenes and the same single cell neuronal architecture. This allows us to concentrate on the feature extraction and neuronal coding properties of these rules. Included in these rules are kurtosis and skewness maximization, the quadratic form of the Bienenstock Cooper-Munro (BCM) learning rule, and single-cell independent component analysis. Using a structure removal method, we demonstrate that receptive fields developed using these rules depend on a small portion of the distribution. We find that the quadratic form of the BCM rule behaves in a manner similar to a kurtosis maximization rule when the distribution contains kurtotic directions, although the BCM modification equations are computationally simpler. PMID- 9744899 TI - Neural feature abstraction from judgments of similarity. AB - The common neural network modeling practice of representing the elements of a task domain in terms of a set of features lacks justification if the features are derived through some form of ad hoc preabstraction. By examining a featural similarity model related to established multidimensional scaling techniques, a neural network is developed that generates features from similarity data and attaches weights to these features. The network performs a constrained search of a continuous solution space to determine the features and uses a previously developed regularization technique to minimize the number of features it derives. The network is demonstrated on artificial data, from which it recovers known features and weights, and on two real data sets involving the similarity of a set of geometric shapes and the abstract conceptual similarities of the 10 Arabic numerals. On the basis of these results, the relationship between the multidimensional scaling approach adopted by the network and an alternative additive clustering approach to feature extraction is discussed. PMID- 9744901 TI - Kernel-Based Equiprobabilistic Topographic Map Formation. AB - We introduce a new unsupervised competitive learning rule, the kernel-based maximum entropy learning rule (kMER), which performs equiprobabilistic topographic map formation in regular, fixed-topology lattices, for use with nonparametric density estimation as well as nonparametric regression analysis. The receptive fields of the formal neurons are overlapping radially symmetric kernels, compatible with radial basis functions (RBFs); but unlike other learning schemes, the radii of these kernels do not have to be chosen in an ad hoc manner: the radii are adapted to the local input density, together with the weight vectors that define the kernel centers, so as to produce maps of which the neurons have an equal probability to be active (equiprobabilistic maps). Both an "online" and a "batch" version of the learning rule are introduced, which are applied to nonparametric density estimation and regression, respectively. The application envisaged is blind source separation (BSS) from nonlinear, noisy mixtures. PMID- 9744900 TI - Classification of temporal patterns in dynamic biological networks. AB - A general method is presented to classify temporal patterns generated by rhythmic biological networks when synaptic connections and cellular properties are known. The method is discrete in nature and relies on algebraic properties of state transitions and graph theory. Elements of the set of rhythms generated by a network are compared using a metric that quantifies the functional differences among them. The rhythms are then classified according to their location in a metric space. Examples are given, and biological implications are discussed. PMID- 9744902 TI - An Energy Function and Continuous Edit Process for Graph Matching. AB - The contributions of this article are twofold. First, we develop a new nonquadratic energy function for graph matching. The starting point is a recently reported mixture model that gauges relational consistency using a series of exponential functions of the Hamming distances between graph neighborhoods. We compute the effective neighborhood potentials associated with the mixture model by identifying the single probability function of zero Kullback divergence. This new energy function is simply a weighted sum of graph Hamming distances. The second contribution is to locate matches by graduated assignment. Rather than solving the mean-field saddle-point equations, which are intractable for our nonquadratic energy function, we apply the soft-assign ansatz to the derivatives of our energy function. Here we introduce a novel departure from the standard graduated assignment formulation of graph matching by allowing the connection strengths of the data graph to update themselves. The aim is to provide a means by which the structure of the data graph can be updated so as to rectify structural errors. The method is evaluated experimentally and is shown to outperform its quadratic counterpart. PMID- 9744903 TI - Approximate Statistical Tests for Comparing Supervised Classification Learning Algorithms. AB - This article reviews five approximate statistical tests for determining whether one learning algorithm outperforms another on a particular learning task. These tests are compared experimentally to determine their probability of incorrectly detecting a difference when no difference exists (type I error). Two widely used statistical tests are shown to have high probability of type I error in certain situations and should never be used: a test for difference of two proportions and a paired-differences t test based on taking several random train-test splits. A third test, a paired-differences t test based on 10-fold cross-validation, exhibits somewhat elevated probability of type I error. A fourth test, McNemar's test, is shown to have low type I error. The fifth test is a new test, 5 x 2 cv, based on five iterations of twofold cross-validation. Experiments show that this test also has acceptable type I error. The article also measures the power (ability to detect algorithm differences when they do exist) of these tests. The cross-validated t test is the most powerful. The 5 x 2 cv test is shown to be slightly more powerful than McNemar's test. The choice of the best test is determined by the computational cost of running the learning algorithm. For algorithms that can be executed only once, McNemar's test is the only test with acceptable type I error. For algorithms that can be executed 10 times, the 5 x 2 cv test is recommended, because it is slightly more powerful and because it directly measures variation due to the choice of training set. PMID- 9744904 TI - Probability Density Estimation Using Entropy Maximization. AB - We propose a method for estimating probability density functions and conditional density functions by training on data produced by such distributions. The algorithm employs new stochastic variables that amount to coding of the input, using a principle of entropy maximization. It is shown to be closely related to the maximum likelihood approach. The encoding step of the algorithm provides an estimate of the probability distribution. The decoding step serves as a generative mode, producing an ensemble of data with the desired distribution. The algorithm is readily implemented by neural networks, using stochastic gradient ascent to achieve entropy maximization. PMID- 9744905 TI - The utility of patch tests using larger screening series of allergens. AB - BACKGROUND: The number of patch test allergens available within the United States for routine commercial purchase is limited. Allergens chosen for inclusion in routine screening series or patch test trays vary, and the degree of information obtained from any series may or may not serve a patient's needs. OBJECTIVE: Knowledge of how well the allergens chosen for inclusion in the two commercially available sources perform compared with a more expansive panel of tests can help physicians select the more appropriate tests. METHODS: From 1994 to mid-1997, 554 patients were tested with allergens recommended by the North American Contact Dermatitis Group (NACDG). This included all allergens currently available from both current domestic sources, although not in the identical form used by the Thin-layer Rapid Use Epicutaneous Test (TRUE) test (Glaxo Dermatology, Research Triangle Park, NC). Another 185 patients were tested with supplemental series of allergens. RESULTS: The larger the series of allergens used, the more positive tests were found and the more relevant tests as well. Hermal patch test allergens identified about 55% of the information found by the NACDG series; the TRUE test allergens (but not in the TRUE test system) identified 65%. Of the 103 reactions to supplemental allergens not found by the NACDG series, 59 were relevant. CONCLUSION: Larger series of allergens can enhance accurate diagnosis of allergic contact dermatitis. No single arbitrary series of allergens can adequately survey the contemporary environment of individual patients. Selection of allergens for testing requires consideration of the patient's history and access to appropriate environmental contactants. PMID- 9744906 TI - The sensitizing capacity of ginkgolic acids in guinea pigs. AB - BACKGROUND: Ginkgo biloba possesses fruits that have caused numerous cases of allergic contact dermatitis. Low amounts of the ginkgolic acids occur in the leaves as well. OBJECTIVE: Leaf extracts are used to treat cerebrovascular and peripheral vascular disorders. The question arises whether skin hypersensitivity reactions may be adverse effects because the pharmaceutical preparations contain low amounts of ginkgolic acids. METHODS: Guinea pigs were sensitized experimentally with pure ginkgolic acids as well as with leaf extracts containing approximately 1,000 ppm of ginkgolic acids. RESULTS: The guinea pigs could be sensitized successfully with the pure ginkgolic acids. The animals could not be sensitized with the leaf extract. CONCLUSION: Leaf extracts of Ginkgo biloba taken orally or given by infusion to treat diffuse cerebral disturbances can be considered safe, even when they might contain up to 1,000 ppm of the sensitizing ginkgolic acids. PMID- 9744907 TI - An economic evaluation of patch testing in the diagnosis and management of allergic contact dermatitis. AB - BACKGROUND: A previous retrospective study indicated that patch testing is cost effective and well accepted by patients. OBJECTIVE: The objective of this observational prospective study was to show the cost-effectiveness of patch testing in patients suspected of allergic contact dermatitis (ACD) and to determine the order in which different severity groups rank in terms of cost effectiveness. METHODS: This observational study was conducted in 567 patients from 10 investigator sites over a period of 1 year. All patients with a suspicion of contact allergy who exhibited at least moderate disease activity were included in the study and were stratified according to disease severity and whether or not they were patch tested. In each severity category, the cost-effectiveness of patch testing was evaluated. Patients who were ruled out for contact allergy by the first 6 months after admission were excluded. A validated dermatology specific quality of life instrument was administered to all the patients at entrance into the study and at 6 and 12 months after that. The cost-effectiveness analysis is shown using a decision analysis model. RESULTS: Patch testing was performed on 22% of patients with mild disease, 41% of patients with moderate disease, and 50% of patients with severe disease. As a result of changes made in their lifestyle, 66% in the patch-tested group and 51% in the non-patch-tested group reported 75% or more improvement in disease symptoms after 6 months. Early confirmation of diagnosis helped reduce the prediagnosis costs of treatment, which was mostly based on preliminary diagnosis. The greatest quality of life benefits from patch testing, relative to no patch testing, occurred in subjects with recurrent or chronic ACD. CONCLUSION: Patch testing is most cost-effective and reduces the cost of therapy in patients with severe ACD. PMID- 9744908 TI - Risk assessment of contact allergens. AB - BACKGROUND: Quantitative risk assessment has been used successfully for a number of human health hazards associated with product use. Dermal contact allergy or sensitization is a relatively late addition to the list. OBJECTIVE: This article outlines the concepts and operational elements of product safety risk assessment of contact allergens. METHODS: The basic risk assessment paradigm is presented along with various dermal dose metrics in the context of their application in describing the dose response or risk of allergic response per unit dose of a product. This analysis of toxic effect is then combined with an assessment of exposure in the real world to estimate the risk of elicitation of active skin disease. RESULTS: Human health risk assessment in general and as it is applied to contact allergy in particular is an emerging science. As with any real science, it allows for the formulation and testing of hypotheses that elucidate a model or description of reality. The model, in turn, allows for predictions that, in this case, are the risk assessments. Of course, the elements of this model and its predictions are also then testable in successive rounds of hypothesis formation and investigation. The entire system runs on data, and a lack of data with a concomitant abundance of scientific uncertainty limits the utility and ultimately the credibility of the process. CONCLUSIONS: Uncertainty from variability and lack of knowledge notwithstanding, the risk assessment paradigm and its scientific construct are valuable in gauging and ultimately controlling the risk of contact allergy from products. PMID- 9744909 TI - Aging and the epidermal permeability barrier: implications for contact dermatitis. AB - OBJECTIVES: This report examines our present knowledge of the epidermal permeability barrier as it pertains to irritant contact dermatitis and discusses how altered barrier function may affect the clinical manifestations of irritant contact dermatitis in the aged. CONCLUSIONS: Altered barrier function affects the ability of an irritant to penetrate the stratum corneum and produce deleterious effects on both stratum corneum and epidermis. Moreover, a damaged epidermis can be expected to produce a defective stratum corneum, thus resulting in a vicious cycle. Furthermore, although basal transepidermal water loss has traditionally been used as an assessment of barrier function, evidence suggests that dynamic tests of barrier function are more reliable indicators of barrier function in the clinical setting of additive, repetitive, or chronic insults. Finally, barrier disruption per se has now been shown to produce both a cytokine response and an increase in epidermal Langerhans' cell density. Thus, barrier disruption not only alters penetrance of contactants, but also may prime the inflammatory response. Studies of aged epidermal permeability barrier function provide a greater understanding of the modified response to irritants in the aged. PMID- 9744910 TI - Factors defining sensitive skin and its treatment. AB - BACKGROUND: Users of cosmetics and skin care products often report adverse reactions ranging from itching and dryness to intense inflammatory responses such as erythema or wheal and rash. Self-assessment is not always an accurate parameter for categorizing skin as sensitive or nonsensitive, although it can be valuable. For this reason, it is important to define sensitive skin by more objective factors. OBJECTIVE: Studies were undertaken to determine if objective biophysical measurements could detect differences in barrier function between those individuals who identified themselves as having sensitive skin and those self-identified as having normal skin. In addition, the effects of treatment on barrier functions of individuals with sensitive skin were determined. METHODS: Three main factors that contribute to cutaneous reactivities were observed for the estimation of skin sensitivity: barrier functions, reactivity to irritants, and neuronal responses manifested as sensory reactions. Barrier functions of the skin was tested by gentle removal of the stratum corneum with simple cellophane tape stripping followed by measurement of transepidermal water loss (TEWL) as a marker of barrier loss. The onset and intensity of skin reaction against an irritant, balsam of Peru, was tested on the same individuals to observe the reactivity of their skin. Using the lactic acid sting test, additional information regarding skin sensitivities was obtained. RESULTS: Sensitive skin individuals exhibiting easy barrier damage possess delicate skin that is also highly reactive to irritants. When these individuals used a regimen of products that contained minimal preservatives and no surfactants for 8 weeks, the skin barrier and reactivity changed such that it was similar to nonsensitive skin. CONCLUSIONS: Skin sensitivity is observed because of a combination of factors, including a disrupted barrier and a tendency to hyperreact to topical agents. Treatment with special topical skin care formulations can reduce overall skin sensitivity. PMID- 9744911 TI - Limits of ICD-9-CM code usefulness in epidemiological studies of contact and other types of dermatitis. AB - BACKGROUND: International Classification of Diseases, Version 9, Clinical Modification (ICD-9-CM) coding information used for billing is readily available in computerized form. OBJECTIVES: The purpose of this article is to determine the usefulness of ICD-9-CM codes in a descriptive dermatoepidemiological study of contact and other dermatitis. METHODS: Prospective recording of specific dermatologic diagnoses and the ICD-9-CM code assigned for each diagnosis was performed for all patient visits to the author's dermatology clinics for 6 months. RESULTS: There were 2,524 patient visits with 4,451 diagnoses, of which 789 diagnoses were dermatitis. The 10 different diagnostic categories of dermatitis had eight associated ICD-9-CM codes. Allergic contact dermatitis with 247 visits, irritant contact dermatitis with 30 visits, and nummular dermatitis with 61 visits shared one diagnostic code. Thus, 43% of visits for dermatitis were intermixed by having the same ICD-9-CM code. CONCLUSION: Lack of one-to-one correspondence of ICD-9-CM codes with dermatitis diagnostic categories creates a situation in which ICD-9-CM codes are not useful for dermatoepidemiological studies of contact and other types of dermatitis. This could be corrected by assigning additional five-digit ICD-9-CM codes to cover each type of dermatitis. Coding for specific allergens or irritants is not feasible with the current five digit ICD-9-CM codes. PMID- 9744912 TI - Occupational dermatitis to ultraviolet-cured acrylic-based inks in computer hard disc manufacturing. AB - OBJECTIVE: Ultraviolet-cured acrylates and their various components and applications are reviewed in this report. METHODS: A 26-year-old woman involved in silk screening computer discs with ultraviolet-cured inks developed an acute allergic contact dermatitis on her hands and forearms. Patch testing revealed a number of strong reactions to epoxy resin and many multifunctional acrylates. RESULTS: The only one listed on the material safety data sheet to which she reacted was tripropyleneglycol diacrylate (TRPGDA). The other positive reactions likely represent cross-reactions. The positive reaction to epoxy is possibly relevant and attributable to nonhardened epoxy resin contaminant in the epoxy prepolymer. CONCLUSION: This case report shows a new application for ultraviolet cured acrylate based inks in computer hard disc manufacturing. PMID- 9744913 TI - Dioxopromethazine-induced photoallergic contact dermatitis followed by persistent light reaction. AB - BACKGROUND: Although photosensitivity after photoallergy to topical phenothiazine antihistamines is well known, there have been no previous reports of dioxopromethazine inducing this phenomenon. OBJECTIVE: A housewife used 0.5% dioxopromethazine in Prothanon gel for palpebral pruritus and developed severe dermatitis of the lower eyelids with spread to the sun-exposed areas. METHODS: The minimal erythema doses and the minimal infiltrate doses for ultraviolet A (UVA) and ultraviolet B (UVB) were established before photopatch testing and at intervals up to 497 days thereafter. Test sites were read up to 144 hours after irradiation. Photopatch testing was performed with Prothanon gel, dioxopromethazine hydrochloride 0.001% to 0.5%, and the standard photopatch test tray (Hermal/Trolab). For patch testing, various series of the German Contact Dermatitis Group were applied. RESULTS: Minimal erythema doses for UVA were diminished before photopatch testing and at intervals up to 500 days after Prothanon gel was discontinued. Exposure to UVB provoked abnormal delayed infiltrated reactions. Clinically the photosensitivity persisted within this period. Photoallergic reactions were seen with Prothanon gel, dioxopromethazine hydrochloride 0.005% to 1.0%, and promethazine hydrochloride 0.1%. The patient gave positive patch test reactions to various fragrance materials, balsam of Peru, costus oil, and propylene glycol. CONCLUSION: Because topical dioxopromethazine may cause photoallergic contact dermatitis followed by long lasting photosensitivity even after contact has been discontinued, its withdrawal from the market is recommended. PMID- 9744914 TI - Occupational allergic contact dermatitis from cyanoacrylate. AB - Cyanoacrylates are widely used in adhesive techniques. Cyanoacrylate adhesives differ physically for the different needs of application, and chemically in function of the size of ester molecules. A 40-year-old man employed at the National Mint and Stamp factory presented with hyperkeratotic lesions on the fingers of the right hand. His job consisted of fixing microchips to plastic phone cards with Loctite Series 414. Patch testing confirmed sensitivity to cyanoacrylates. After the diagnosis of allergic contact dermatitis was established, the patient, to be cured of the dermatitis, changed his workplace 2 months later. PMID- 9744916 TI - Masking fragrances revisited. PMID- 9744915 TI - Airborne occupational allergic contact dermatitis from champignon mushroom. AB - BACKGROUND: Allergic contact dermatitis from mushrooms has only seldom been reported. OBJECTIVES: We report on a mushroom picker who developed skin symptoms from occupational exposure to the mushroom champignon. METHODS: Conventional patch testing and prick testings were performed. RESULTS: Erythema and vesicular edemic dermatitis appeared around the eyes, on the cheeks, around the nose, and around the lips of a 31-year-old woman who had been involved in the commercial production of champignons for 5 years. Prick testing to champignon was negative, but patch testing with raw champignon provoked a 2+ allergic reaction and was negative in the controls. CONCLUSION: Our patient had been occupationally sensitized from exposure to champignon. The allergen is not known but may be a low-molecularweight chemical or a protein present in the champignon. PMID- 9744917 TI - Contact leukoderma caused by patch testing with dental acrylics. AB - Several chemicals are capable of inducing contact leukoderma. Here we report on a dental nurse who had been investigated elsewhere at a dermatology clinic 2.8 years earlier because of suspected occupational fingertip dermatitis. She had been patch tested on her upper arm with dental acrylic resins "as is." These strong concentrations of patch test substances caused a severe allergic reaction in the upper arm, and the patch test sites have remained vitiliginous for 2.8 years. Active sensitization did not take place because the patient had been sensitized earlier as shown by the allergic 2-day readings with acrylics during the first patch test session. It is assumed that acrylates induced contact vitiligo, but the dental acrylics may have also contained other chemicals (eg, hydroquinone or phenolic substances) capable of causing vitiligo. The main point to be learned from the present results is that dental acrylics should never be patch tested "as is." We also discourage the practice of use tests, open tests, or repeated open patch tests with undiluted dental acrylics because of the risk of active sensitization from single exposure. PMID- 9744918 TI - Suppression of NMDA receptor function using antisense DNA block ocular dominance plasticity while preserving visual responses. AB - Pioneering work has shown that pharmacological blockade of the N-methyl-D aspartate (NMDA) receptor channel reduces ocular dominance plasticity. However, the results also show that doses of NMDA receptor antagonists that have an effect on ocular dominance plasticity profoundly reduce sensory responses and disrupt stimulus selectivity of cortical cells. It is, therefore, not possible to determine whether effects of NMDA receptor blockade on visual plasticity result from a specific role of NMDA receptors or from the reduction in sensory response. We have used an alternate approach to examine this question. We performed knockdown experiments using antisense oligodeoxynucleotides (ODNs) complementary to mRNA coding the NR1 subunit of the NMDA receptor. After 5 days of antisense, but not sense, ODN treatment NMDA receptor-mediated synaptic transmission was reduced markedly relative to the alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid (AMPA) receptor response, as indicated by whole cell patch-clamp recordings in the cortical slice preparation. This suppression of NMDA receptor-mediated currents was due to a selective reduction in the NR1 protein near the injection site relative to the untreated hemisphere in the same animal, as indicated by immunocytochemistry and Western blotting. In contrast, AMPA receptors were not affected by the antisense ODN treatment indicating specificity of effects. Another major effect of this treatment was to decrease ocular dominance plasticity. Ferrets that were monocularly deprived 1 wk during the antisense ODN treatment had ocular dominance histograms similar to those found in untreated, nondeprived animals. In contrast, ferrets treated with sense ODN and monocularly deprived had ocular dominance histograms resembling those of untreated, monocularly deprived animals. The effects on ocular dominance plasticity did not result from a disruption of sensory responses because maximum responses as well as orientation and direction selectivity of cortical cells were not affected by the treatment. In conclusion, the present results show that antisense techniques can accomplish more selective manipulations of cortical function than is possible with traditional pharmacological agents. Use of this approach also provides unambiguous evidence for a specific role of NMDA receptors in visual plasticity. PMID- 9744919 TI - 5-Hydroxytryptamine responses in immature rat rostral ventrolateral medulla neurons in vitro. AB - Whole cell patch recordings were made from rostral ventrolateral medullar (RVLM) neurons of brain-stem slices from 8- to 12-day-old rats. By superfusion or pressure ejection to RVLM neurons, 5-hydroxytryptamine (5-HT) elicited three types of membrane potential changes: a slow hyperpolarization (5-HTH), a slow depolarization (5-HTD) and a biphasic response, which persisted in a tetrodotoxin (TTX, 0.3 microM)-containing solution. 5-HTH were accompanied by a decrease of input resistance in the majority of responsive neurons. Hyperpolarization reduced and depolarization increased the 5-HTH; the mean reversal potential was -92.3 mV in 3.1 mM and shifted to -69.3 mV in 7 mM [K+]o. Barium (Ba2+, 0.1 mM) but not tetraethylammonium (TEA, 10 mM) suppressed 5-HTH. The 5-HT1A receptor agonist (+/ )-8-hydroxy-dipropylamino-tetralin (8-OH-DPAT; 5-50 microM) hyperpolarized RVLM neurons. The 5-HT1A antagonist pindobind-5-HT1A (PBD; 1-3 microM) and the 5-HT2/5 HT1 receptor antagonist spiperone (1-10 microM) suppressed 5-HTH and the hyperpolarizing phase of biphasic responses; the 5-HT2 receptor antagonist ketanserin (3 microM) was without significant effect. 5-HTD were associated with an increase or no apparent change of input resistance in RVLM neurons. Hyperpolarization of the membrane decreased or caused no apparent change in 5 HTD. 5-HTD were reduced in an elevated [K+]o (7.0 mM) solution and > 60% in a low Na+ (26 mM) solution and were not significantly changed in a low Cl- (6.7 mM) or Ca(2+)-free/high Mg2+ (10.9 mM) solution. The 5-HT2 receptor agonist alpha-methyl 5-HT (50 microM) depolarized RVLM neurons, and the 5-HT2 antagonist ketanserin (1 10 microM) attenuated the 5-HTD and the depolarizing phase of biphasic responses, whereas the 5-HT1A receptor antagonist PBD (2 microM) was without effect. Inclusion of the hydrolysis resistant guanine nucleotide GDP-beta-S in patch solution significantly reduced the 5-HTH as well as the 5-HTD. The present study shows that, in the immature rat RVLM neurons, 5-HT causes a slow hyperpolarization and depolarization probably by interacting with 5-HT1A and 5 HT2 receptors, which are G-proteins coupled. 5-HTH may involve an increase of an inwardly rectifying K+ conductance, and 5-HTD appear to be caused by a decrease of K+ conductance and/or increase of nonselective cation conductance. PMID- 9744920 TI - Characterization of carbachol-induced rhythmic bursting discharges in neurons from guinea pig lateral septum slices. AB - A brain slice preparation from guinea pigs and intracellular recording techniques were used to examine the effects of carbachol application on the three classes (A, B, and C) of neurons (n = 68, 40 of class A, 12 of class B, 16 of class C) within the mediolateral part of the lateral septum (LSml). Bath application of carbachol elicited a sustained depolarization associated with an increase in membrane input resistance, action-potential firing and triggered rhythmic bursting discharges in 59% of recorded neurons. According to the configuration of these bursts, LSml neurons were classified into type I, II, and III neurons with reference to their response to carbachol. The frequency of spontaneous bursts was increased by depolarization caused by applied DC current in the three types of neurons. Bursts in type II and III neurons were voltage and dose dependent. These dependences were responsible for a continuum of variation in carbachol responses in these two types of neurons. As the neuron depolarized in the presence of carbachol, spontaneous action potentials increased in frequency and slow afterdepolarizing potentials (sADPs) appeared and preceded the occurrence of the first burst. These sADPs from adjacent action potentials appeared to progressively increase to initiate a burst. In the presence of carbachol, sADPs and bursts were also observable after action potentials evoked by depolarizing current pulses at the resting membrane potential (RMP) in LSml neurons. Evoked sADPs and bursts were associated with an apparent increase in input conductance. Application of low Na+ medium blocked both the sADP and bursts. Application of zero Ca2+ medium either 1) blocked completely the generation of sADPs and bursts (n = 16), or 2) did not block bursts (n = 14). Evoked sADPs and bursts were blocked by tetraethylammonium but were resistant to external Cs+. The results indicate that the activation of cholinergic receptors does not differentially affect the three classes of LSml neurons. The responses to carbachol in type II and III neurons form a continuum of variation, whereas these of type I neurons constitute a discrete entity. The selective cholinergic induction of a sADP, and the progressive activation of these sADPs in LSml neurons are thought to be responsible for the onset of the three types of rhythmic bursting discharges. We propose that sADPs and bursts induced by carbachol are generated by a cationic conductance largely permeable to Na+. In a subpopulation of LSml neurons (n = 16), the bursts are dependent on the presence of Ca2+ in the medium. PMID- 9744921 TI - Passive transfer of Lambert-Eaton myasthenic syndrome induces dihydropyridine sensitivity of ICa in mouse motor nerve terminals. AB - Mice were injected for 30 days with plasma from three patients with Lambert-Eaton Myasthenic Syndrome (LEMS). Recordings were made from the perineurial sheath of motor axon terminals of triangularis sterni muscle preparations. The objective was to characterize pharmacologically the identity of kinetically distinct, defined potential changes associated with motor nerve terminal Ca2+ currents (ICa) that were affected by LEMS autoantibodies. ICa elicited at 0.01 Hz were significantly reduced in amplitude by approximately 35% of control in LEMS treated nerve terminals. During 10-Hz stimulation, ICa amplitude was unchanged in LEMS-treated motor nerve terminals, but was depressed in control. During 20- or 100-Hz trains, facilitation of ICa occurred in LEMS-treated nerve terminals whereas in control, no facilitation occurred during the trains at 20 Hz and marked depression occurred at 100 Hz. Saturation for amplitude and duration of ICa in control terminals occurred at 2 and 4-6 mM extracellular Ca2+, respectively; in LEMS-treated terminals, the extracellular Ca2+ concentration had to increase by two to three times of control to cause saturation. Amplitude of the two components of ICa observed when the preparation was exposed to 50 microM 3,4-diaminopyridine and 1 mM tetraethylammonium were both reduced by LEMS plasma treatment. The fast component (ICa,s) was reduced by 35%, whereas the slow component (ICa, s) was reduced by 37%. omega-Agatoxin IVA (omega-Aga-IVA; 0.15 microM) and omega-conotoxin-MVIIC (omega-CTx-MVIIC; 5 microM) completely blocked ICa in control motor nerve terminals. The same concentrations of toxins were 20 30% less effective in blocking ICa in LEMS-treated terminals. The residual ICa remaining after treatment with omega-Aga-IVA or omega-CTx-MVIIC was blocked by 10 microM nifedipine and 10 microM Cd2+. Thus LEMS plasma appears to downregulate omega-Aga-IVA-sensitive (P-type) and/or omega-CTx-MVIIC-sensitive (Q-type) Ca2+ channels in murine motor nerve terminals, whereas dihydropyridine (DHP)-sensitive (L-type) Ca2+ channels are unmasked in these terminals. Acute exposure (90 min) of rat forebrain synaptosomes to LEMS immunoglobulins (Igs; 4 mg/ml) did not alter the binding of [3H]-nitrendipine or [125I]-omega-conotoxin-GVIA (-omega CgTx GVIA) when compared with synaptosomes incubated with an equivalent concentration of control Igs. Conversely, LEMS Igs significantly decreased the Bmax for [3H]-verapamil to approximately 45% of control. The apparent affinity of verapamil (KD) for the remaining receptors was not significantly altered. Thus acute exposure of isolated central nerve terminals to LEMS Igs does not increase DHP sensitivity, whereas it reduces the number of binding sites for verapamil but not for nitrendipine or omega-CgTx-GVIA. These results suggest that chronic but not acute exposure to LEMS Igs either upregulates or unmasks DHP-sensitive Ca2+ channels in motor nerve endings. PMID- 9744922 TI - Contrast enhancement and distributed encoding by bipolar cells in the retina. AB - Responses of bipolar cells, cone photoreceptors, and horizontal cells were recorded intracellularly in superfused eyecup preparations of the tiger salamander (Ambystoma tigrinum). Contrast flashes of positive and negative polarity were applied at the center of the receptive field while the entire retina was light adapted to a background field of 20 cd/m2. For small contrasts, many bipolar cells showed remarkably high contrast gain: up to 15-20% of the bipolar response was evoked by a contrast step of 1%. There was considerable variation from cell to cell but, on average, no striking differences in contrast gain were found between the depolarizing (Bd) and hyperpolarizing (Bh) bipolar cells. Quantitative comparisons of contrast/response measurements for cone photoreceptors and cone-driven bipolars suggest that the high contrast gain of bipolars is the consequence of a 5-10 x amplification of small signals across the cone-->bipolar synapse. Bipolar cells had a very restricted linear range of response and tended to saturate at stimulus levels that were within the linear range of the cone response. The contrast/response of horizontal cells was similar to that of cones and differed markedly from that of Bh cells. For steps of equal contrast, the latency of the Bh cells was approximately 20 ms shorter than that of the Bd cells regardless of the contrast magnitude. For both bipolar cells and cones, the effect of contrast polarity on latency seems largely due to the absolute value of the light step, delta L. In the large signal domain, properties of the contrast responses of bipolar cells varied appreciably, both within and between the Bd and Bh classes. Cells of either class could be positive- or negative-contrast dominant. These and additional results show that in the light adapted retina, the bipolar population is functionally diverse and has the potential to provide a rich substrate for distributed encoding of visual images. PMID- 9744923 TI - Myelinated mechanically insensitive afferents from monkey hairy skin: heat response properties. AB - To compare the heat responses of mechanically sensitive and mechanically insensitive A-fiber nociceptors, an electrical search technique was used to locate the receptive fields of 156 A-fibers that innervated the hairy skin in the anesthetized monkey (77 A beta-fibers, 79 A delta-fibers). Two-thirds of these afferents were either low-threshold mechanoreceptors (n = 91) or low-threshold cold receptors (n = 11). Nine A beta-fibers and 41 A delta-fibers were cutaneous nociceptors, and four A delta-fibers innervated subcutaneous tissue. The majority of cutaneous A-fiber nociceptors were heat sensitive (43/50 = 86%). Heat insensitive cutaneous A-fiber nociceptors consisted of one cold nociceptor, three silent nociceptors, and three high-threshold mechanoreceptors. Two types of response were observed to an intense heat stimulus (53 degrees C, 30 s). Type I (n = 26) was characterized by a long latency (mean: 5 s) and a late peak discharge (16 s). Type II (n = 17) was characterized by a short latency (0.2 s) and an early peak discharge (0.5 s). Type I fibers exhibited faster conduction velocities (25 vs. 14 m/s) and higher heat thresholds (> 53 vs. 47 degrees C, 1-s duration) than type II fibers. The possibility that the type I heat response was a result of sensitization was tested in three fibers by determining the heat threshold to 30-s duration stimuli (42-46 degrees C). For this long stimulus duration heat thresholds were reproducible across multiple runs, and the threshold to the 1-s duration stimulus was not altered by these tests. Thus fibers with a type I heat response were not high-threshold mechanoreceptors that developed a heat response through sensitization. Fibers with a type II heat response had significantly higher mechanical thresholds (median: 15 bar) than fibers with a type I heat response (5 bar). This finding accounts for the observation that type II heat responses were infrequently observed in earlier studies wherein the search technique depended on mechanical responsiveness. Fibers with a type II response exhibited a graded response to heat stimuli, marked fatigue to repeated applications of heat stimuli, and adaptation to sustained heat stimuli similar to that seen in C-fiber nociceptors. First pain sensation to heat is served by type II A-fiber nociceptors that are mechanically insensitive. Type I A-fiber nociceptors likely signal pain to long-duration heat stimuli and may signal first pain sensation to mechanical stimuli. PMID- 9744924 TI - Inhibition of a hyperpolarization-activated current by clonidine in rat dorsal root ganglion neurons. AB - Whole cell voltage- and current-clamp recordings were carried out to investigate the effects of clonidine, an alpha 2-adrenoceptor agonist, in L4 and L5 dorsal root ganglion (DRG) neurons of the rat. In voltage-clamp mode, application of 20 microM clonidine reversibly reduced the inward current evoked by hyperpolarizing voltage steps. The "clonidine-sensitive current" was obtained by subtracting the current during clonidine application from the control current, and its properties were as follows. 1) It was a slowly activating inward current evoked by hyperpolarization. 2) The reversal potential in the standard extracellular solution ([K+]o = 5 mM, [Na+]o = 151 mM) was -38.3 mV, and reduction of [Na+]o shifted it to a more negative potential, whereas an increase of [K+]o shifted it to a more positive potential, indicating that the current was carried by Na+ and K+ (PNa/PK = 0.22). 3) The relationship between the chord conductance underlying the clonidine-sensitive current and voltage could be fitted by a Boltzmann equation. These results indicate that the clonidine-sensitive current corresponds to a hyperpolarization-activated current (Ih), i.e., clonidine inhibits Ih in rat DRG neurons. DRG neurons were classified as small (15.9-32.9 microns diam), medium-sized (33-42.9 microns), and large (43-63.6 microns), and 7 of 19, 24 of 25, and 22 of 22 of these types exhibited Ih with mean +/- SE clonidine-induced inhibition values of 36.1 +/- 3.5% (n = 7), 43.1 +/- 3.7% (n = 24), and 35.1 +/- 2.7% (n = 22), respectively. Clonidine application to L4 and L5 DRG neurons excised from rats the sciatic nerves of which had been transected 14-35 days previously (transected DRG neurons) also reduced Ih. In current-clamp mode, 9 of 13 intact and 4 of 6 transected medium-sized DRG neurons that exhibited Ih responded to clonidine with hyperpolarization (> 2 mV). Some medium-sized DRG neurons exhibited repetitive action potentials in response to a depolarizing current pulse, and clonidine reduced the firing discharge frequencies in 8 of 11 intact and 3 of 4 transected neurons tested. Injection of a hyperpolarizing current pulse produced time-dependent rectification in DRG neurons that exhibited Ih, and clonidine blocked this rectification in all intact and transected neurons tested. These results suggest that inhibition of Ih due to alpha 2-adrenoceptor activation contributes to modulation of DRG neuronal activity in rats. On the basis of our findings, we discuss the possible mechanisms whereby sympathetically released norepinephrine modulates the abnormal activity of DRG neuronal cell bodies after nerve injury. PMID- 9744926 TI - Serotonin reduces polysynaptic inhibition via 5-HT1A receptors in the superficial entorhinal cortex. AB - The superficial cells of the entorhinal cortex (EC), main input to the hippocampus, receive a serotonergic input from the raphe nuclei and express 5 hydroxytryptamine creatine sulfate complex (5-HT) receptors at high density. With the use of intracellular recordings, we investigated the effects of serotonin on synaptic inhibition of layer II and III neurons of the EC. Serotonin reduced both polysynaptic fast and slow inhibitory postsynaptic potentials (IPSPs) in projection neurons of the superficial EC. Polysynaptic fast and slow IPSPs were depressed by serotonin in a dose-dependent manner (0.1-100 microM). Serotonin in a concentration of 1 microM reduced the amplitudes of polysynaptic fast and slow IPSPs by approximately 40 and 50%, respectively. To identify the subtype of the 5 HT-receptor mediating the effects on polysynaptic IPSPs, we applied various 5-HT receptor agonists and antagonists. Although the serotonin agonists for the 5 HT1B,2C,3 receptors were ineffective, the effects were mimicked by the 5-HT1A receptor agonists (8-OH-DPAT, 5-CT) and prevented by the 5-HT1A-receptor antagonist NAN-190. To look at the direct effects of 5-HT on inhibitory interneurons, we elicited monosynaptic IPSPs in the absence of excitatory synaptic transmission. In contrast to the polysynaptic IPSPs, monosynaptic IPSPs were not significantly affected by serotonin. Recordings from putative inhibitory interneurons revealed that their excitatory postsynaptic potentials (EPSPs) were reversibly reduced by serotonin. We conclude that serotonin suppresses polysynaptic inhibition in projection neurons of layers II and III of the EC by depression of EPSPs on inhibitory interneurons via 5-HT1A receptors. PMID- 9744927 TI - Control of recurrent inhibition of the lateral posterior-pulvinar complex by afferents from the deep layers of the superior colliculus of the rabbit. AB - We investigated the effect of stimulation of the deep layers of the superior colliculus (SC) on the recurrent inhibition of the lateral posterior-pulvinar complex (LP) in anesthetized rabbits. Intracellular recordings from 23 relay cells in LP showed that they responded to SC stimulation with a long-lasting (140.2 +/- 19.6 ms; mean +/- SD) inhibitory postsynaptic potential (IPSP), which sometimes was followed by a rebound burst of spikes. The same SC stimulation evoked a burst of spikes in extracellular recordings from 31 recurrent inhibitory interneurons in the LP-cortical pathway, which were located in the ventral part of the visual sector of the thalamic reticular nucleus. The mean latency of the burst in reticular cells was 1.6 ms shorter than that of the IPSP in LP relay cells, suggesting that the IPSP in LP cells was mediated by these reticular cells. Intracellular recordings from nine reticular cells showed that the burst of spikes evoked by SC stimulation resulted from an excitatory postsynaptic potential that was always followed by a long-lasting (143.3 +/- 24.0 ms) IPSP. Stimulation of the contralateral predorsal bundle, the main output pathway of deep SC neurons, elicited similar responses in LP cells or reticular neurons with latencies longer than those from SC stimulation. The latency difference between the responses to predorsal bundle and SC stimulation is equal to the antidromic conduction time of predorsal bundle fibers, suggesting that the inhibition in LP originates from the activation of predorsal bundle-projecting neurons. The response characteristics of the inhibitory circuit of LP and of the lateral geniculate nucleus to SC stimulation are strikingly similar, implying that a similar circuit is used by predorsal bundle-projecting neurons to control the recurrent inhibition in both lateral geniculate nucleus and LP. Because the predorsal bundle-projecting neurons are believed to be involved in the initiation of saccadic eye movements, we suggest that the inhibitory circuits may play an important role in modulating ascending visual information during saccadic eye movements. PMID- 9744925 TI - Calcium released from intracellular stores inhibits GABAA-mediated currents in ganglion cells of the turtle retina. AB - We studied spiking neurons isolated from turtle retina by the whole cell version of the patch clamp. The studied cells had perikaryal diameters > 15 microns and fired multiple spikes in response to depolarizing current steps, indicating they were ganglion cells. In symmetrical [Cl-], currents elicited by puffs of 100 microM gamma-aminobutyric acid (GABA) were inward at a holding potential of -80 mV. All of the GABA-evoked current was blocked by SR95331 (20 microM), indicating that it was mediated by a GABAA receptor. The GABA-evoked currents were unaltered by eliciting a transmembrane calcium current either just before or during the response to GABA. On the other hand caffeine (10 mM), which induces Ca2+ release from intracellular stores, inhibited the GABA-evoked current on average by 30%. The caffeine effect was blocked by introducing the calcium buffer bis-(o aminophenoxy)-N,N,N',N'-tetraacetic acid (BAPTA) into the cell but was unaffected by replacing [Ca2+]o with equimolar cobalt. Thapsigargin (10 microM), an inhibitor of intracellular calcium pumps, and ryanodine (20 microM), which depletes intracellular calcium stores, both markedly reduced a caffeine-induced inhibition of the GABA-evoked current. Another activator of intracellular calcium release, inositol trisphosphate (IP3; 50 microM), also progressively reduced the GABA-induced current when introduced into the cell. Dibutyryl adenosine 3'5' cyclic monophosphate (cAMP; 0.5 mM), a membrane-permeable analogue of cAMP, did not reduce GABA-evoked currents, suggesting that cAMP-dependent kinases are not involved in suppressing GABAA currents, whereas calmidazolium (30 microM) and cyclosporin A (20 microM), which inhibit Ca/calmodulin-dependent phosphatases, did reduce the caffeine-induced inhibition of the GABA-evoked current. Alkaline phosphatase (150 micrograms/ml) and calcineurin (300 micrograms/ml) had a similar action to caffeine or IP3. Antibodies directed against the ryanodine receptor or the IP3 receptor reacted with the great majority of neurons in the ganglion cell layer. We found that these two antibodies colocalized in large ganglion cells. In summary, intracellular calcium plays a role in reducing the currents elicited by GABA, acting through GABAA receptors. The modulatory action of calcium on GABA responses appears to work through one or more Ca-dependent phosphatases. PMID- 9744928 TI - Eye position effects on the neuronal activity of dorsal premotor cortex in the macaque monkey. AB - Visual inputs to the brain are mapped in a retinocentric reference frame, but the motor system plans movements in a body-centered frame. This basic observation implies that the brain must transform target coordinates from one reference frame to another. Physiological studies revealed that the posterior parietal cortex may contribute a large part of such a transformation, but the question remains as to whether the premotor areas receive visual information, from the parietal cortex, readily coded in body-centered coordinates. To answer this question, we studied dorsal premotor cortex (PMd) neurons in two monkeys while they performed a conditional visuomotor task and maintained fixation at different gaze angles. Visual stimuli were presented on a video monitor, and the monkeys made limb movements on a panel of three touch pads located at the bottom of the monitor. A trial begins when the monkey puts its hand on the central pad. Then, later in the trial, a colored cue instructed a limb movement to the left touch pad if red or to the right one if green. The cues lasted for a variable delay, the instructed delay period, and their offset served as the go signal. The fixation spot was presented at the center of the screen or at one of four peripheral locations. Because the monkey's head was restrained, peripheral fixations caused a deviation of the eyes within the orbit, but for each fixation angle, the instructional cue was presented at nine locations with constant retinocentric coordinates. After the presentation of the instructional cue, 133 PMd cells displayed a phasic discharge (signal-related activity), 157 were tonically active during the instructed delay period (set-related or preparatory activity), and 104 were active after the go signal in relation to movement (movement-related activity). A large proportion of cells showed variations of the discharge rate in relation to limb movement direction, but only modest proportions were sensitive to the cue's location (signal, 43%; set, 34%; movement, 29%). More importantly, the activity of most neurons (signal, 74%; set, 79%; movement, 79%) varied significantly (analysis of variance, P < 0.05) with orbital eye position. A regression analysis showed that the neuronal activity varied linearly with eye position along the horizontal and vertical axes and can be approximated by a two-dimensional regression plane. These data provide evidence that eye position signals modulate the neuronal activity beyond sensory areas, including those involved in visually guided reaching limb movements. Further, they show that neuronal activity related to movement preparation and execution combines at least two directional parameters: arm movement direction and gaze direction in space. It is suggested that a substantial population of PMd cells codes limb movement direction in a head-centered reference frame. PMID- 9744929 TI - Human horizontal vestibulo-ocular reflex initiation: effects of acceleration, target distance, and unilateral deafferentation. AB - The vestibulo-ocular reflex (VOR) generates compensatory eye movements in response to angular and linear acceleration sensed by semicircular canals and otoliths respectively. Gaze stabilization demands that responses to linear acceleration be adjusted for viewing distance. This study in humans determined the transient dynamics of VOR initiation during angular and linear acceleration, modification of the VOR by viewing distance, and the effect of unilateral deafferentation. Combinations of unpredictable transient angular and linear head rotation were created by whole body yaw rotation about eccentric axes: 10 cm anterior to eyes, centered between eyes, centered between otoliths, and 20 cm posterior to eyes. Subjects viewed a target 500, 30, or 15 cm away that was extinguished immediately before rotation. There were four stimulus intensities up to a maximum peak acceleration of 2,800 degrees/s2. The normal initial VOR response began 7-10 ms after onset of head rotation. Response gain (eye velocity/head velocity) for near as compared with distant targets was increased as early as 1-11 ms after onset of eye movement; this initial effect was independent of linear acceleration. An otolith mediated effect modified VOR gain depending on both linear acceleration and target distance beginning 25-90 ms after onset of head rotation. For rotational axes anterior to the otoliths, VOR gain for the nearest target was initially higher but later became less than that for the far target. There was no gain correction for the physical separation between the eyes and otoliths. With lower acceleration, there was a nonlinear reduction in the early gain increase with close targets although later otolith mediated effects were not affected. In subjects with unilateral vestibular deafferentation, the initial VOR was quantitatively normal for rotation toward the intact side. When rotating toward the deafferented side, VOR gain remained less than half of normal for at least the initial 55 ms when head acceleration was highest and was not modulated by target distance. After this initial high acceleration period, gain increased to a degree depending on target distance and axis eccentricity. This behavior suggests that the commissural VOR pathways are not modulated by target distance. These results suggest that the VOR is initially driven by short latency ipsilateral target distance dependent and bilateral target-distance independent canal pathways. After 25 ms, otolith inputs contribute to the target distance dependent pathway. The otolith input later grows to eventually dominate the target distance mediated effect. When otolith input is unavailable the target distance mediated canal component persists. Modulation of canal mediated responses by target distance is a nonlinear effect, most evident for high head accelerations. PMID- 9744930 TI - Kinetic and stochastic properties of a persistent sodium current in mature guinea pig cerebellar Purkinje cells. AB - Whole cell voltage-clamp techniques were employed to characterize the sodium (Na) conductances in acutely dissociated, mature guinea-pig cerebellar Purkinje cells. Three phenomenological components were noted: two inactivating and a persistent component (I(P)(Na). All exhibited similar sensitivities to tetrodotoxin (TTX; IC50 approximately 3 nM). The inactivating Na current demonstrates two components with different rates of inactivation. The persistent component activates at a more negative membrane potential than the inactivating components and shows little inactivation during a 5-s pulse. The amplitude of the persistent Na conductance had a higher Q10 than the inactivating Na conductance (2.7 vs. 1.3). (I(P)(Na) rapidly activates (approximately 1 ms) and deactivates (< 0.2 ms) and like the fast component appears to be exclusively Na permeable. (I(P)(Na) is not a "window" current because its range of activation exceeds the small overlap between the steady-state activation and inactivation characteristics of the inactivating current. Anomalous tail currents were observed during voltage pulses above -40 mV after a prepulse above -30 mV. The tails rose to a maximum inward current with a time constant of 1.5 ms and decayed to a persistent inward current with a time constant of 20 ms. The tails probably arose as a result of recovery from inactivation through the open state. The noise characteristics of (I(P)(Na) were anomalous in that the measured variance was lower at threshold voltages than would be predicted by a binomial model. The form of the variance could be partially accounted for by postulating that the maximum probability of activation of the persistent current was less than unity. The noise characteristics of (I(P)(Na) are such as to minimize noise near spike activation threshold and sharpen the threshold. PMID- 9744931 TI - Contribution of the rostral fastigial nucleus to the control of orienting gaze shifts in the head-unrestrained cat. AB - The implication of the caudal part of the fastigial nucleus (cFN) in the control of saccadic shifts of the visual axis is now well established. In contrast a possible involvement of the rostral part of the fastigial nuceus (rFN) remains unknown. In the current study we investigated in the head-unrestrained cat the contribution of the rFN to the control of visually triggered saccadic gaze shifts by measuring the deficits after unilateral muscimol injection in the rFN. A typical gaze dysmetria was observed: gaze saccades directed toward the inactivated side were hypermetric, whereas those with an opposite direction were hypometric. For both movement directions, gaze dysmetria was proportional to target retinal eccentricity and could be described as a modified gain in the translation of visual signals into eye and head motor commands. Correction saccades were triggered when the target remained visible and reduced the gaze fixation error to 2.7 +/- 1.3 degrees (mean +/- SD) on average. The hypermetria of ipsiversive gaze shifts resulted predominantly from a hypermetric response of the eyes, whereas the hypometria of contraversive gaze shifts resulted from hypometric responses of both eye and head. However, even in this latter case, the eye saccade was more affected than the motion of the head. As a consequence, for both directions of gaze shift the relative contributions of the eye and head to the overall gaze displacement were altered by muscimol injection. This was revealed by a decreased contribution of the head for ipsiversive gaze shifts and an increased head contribution for contraversive movements. These modifications were associated with slight changes in the delay between eye and head movement onsets. Inactivation of the rFN also affected the initiation of eye and head movements. Indeed, the latency of ipsiversive gaze and head movements decreased to 88 and 92% of normal, respectively, whereas the latency of contraversive ones increased to 149 and 145%. The deficits induced by rFN inactivation were then compared with those obtained after muscimol injection in the cFN of the same animals. Several deficits differed according to the site of injection within the fastigial nucleus (tonic orbital eye rotation, hypermetria of ipsiversive gaze shifts and fixation offset, relationship between dysmetria and latency of contraversive gaze shifts, postural deficit). In conclusion, the present study demonstrates that the rFN is involved in the initiation and the control of combined eye-head gaze shifts. In addition our findings support a functional distinction between the rFN and cFN for the control of orienting gaze shifts. This distinction is discussed with respect to the segregated fastigiofugal projections arising from the rFN and cFN. PMID- 9744933 TI - Stiffness control of balance in quiet standing. AB - Our goal was to provide some insights into how the CNS controls and maintains an upright standing posture, which is an integral part of activities of daily living. Although researchers have used simple performance measures of maintenance of this posture quite effectively in clinical decision making, the mechanisms and control principles involved have not been clear. We propose a relatively simple control scheme for regulation of upright posture that provides almost instantaneous corrective response and reduces the operating demands on the CNS. The analytic model is derived and experimentally validated. A stiffness model was developed for quiet standing. The model assumes that muscles act as springs to cause the center-of-pressure (COP) to move in phase with the center-of-mass (COM) as the body sways about some desired position. In the sagittal plane this stiffness control exists at the ankle plantarflexors, in the frontal plane by the hip abductors/adductors. On the basis of observations that the COP-COM error signal continuously oscillates, it is evident that the inverted pendulum model is severely underdamped, approaching the undamped condition. The spectrum of this error signal is seen to match that of a tuned mass, spring, damper system, and a curve fit of this "tuned circuit" yields omega n the undamped natural frequency of the system. The effective stiffness of the system, Ke, is then estimated from Ke = I omega n2, and the damping B is estimated from B = BW X I, where BW is the bandwidth of the tuned response (in rad/s), and I is the moment of inertia of the body about the ankle joint. Ten adult subjects were assessed while standing quietly at three stance widths: 50% hip-to-hip distance, 100 and 150%. Subjects stood for 2 min in each position with eyes open; the 100% stance width was repeated with eyes closed. In all trials and in both planes, the COP oscillated virtually in phase (within 6 ms) with COM, which was predicted by a simple 0th order spring model. Sway amplitude decreased as stance width increased, and Ke increased with stance width. A stiffness model would predict sway to vary as Ke 0.5. The experimental results were close to this prediction: sway was proportional to Ke(-0.55). Reactive control of balance was not evident for several reasons. The visual system does not appear to contribute because no significant difference between eyes open and eyes closed results was found at 100% stance width. Vestibular (otolith) and joint proprioceptive reactive control were discounted because the necessary head accelerations, joint displacements, and velocities were well below reported thresholds. Besides, any reactive control would predict that COP would considerably lag (150-250 ms) behind the COM. Because the average COP was only 4 ms delayed behind the COM, reactive control was not evident; this small delay was accounted for by the damping in the tuned mechanical system. PMID- 9744932 TI - Ionic mechanism of the slow afterdepolarization induced by muscarinic receptor activation in rat prefrontal cortex. AB - The mammalian prefrontal cortex receives a dense cholinergic innervation from subcortical regions. We previously have shown that cholinergic stimulation of layer V pyramidal neurons of the rat prefrontal cortex results in a depolarization and the appearance of a slow afterdepolarization (sADP). In the current report we examine the mechanism underlying the sADP with the use of sharp microelectrode and whole cell recording techniques in in vitro brain slices. The ability of acetylcholine (ACh) and carbachol to induce the appearance of an sADP in pyramidal cells of layer V of prefrontal cortex is antagonized in a surmountable manner by atropine and is mimicked by application of muscarine or oxotremorine. These results indicate that ACh acts on muscarinic receptors to induce the sADP. In many cell types afterpotentials are triggered by calcium influx into the cell. Therefore we examined the possibility that calcium influx might be the trigger for the generation of the sADP. Consistent with this possibility, buffering intracellular calcium reduced or abolished the sADP but had little effect on the direct muscarinic receptor-induced depolarization also seen in these cells. These results, coupled to the previous observation that calcium channel blockers inhibit the sADP, indicated that the sADP results from a rise in intracellular calcium secondary to calcium influx into the cell. The ionic basis for the current underlying the sADP (IsADP) was examined with the use of ion substitution experiments. The amplitude of IsADP was found to be reduced in a graded fashion by replacement of extracellular sodium with N-methyl-D glucamine (NMDG). In contrast no clear evidence for the involvement of potassium or chloride channels in the generation of the sADP or IsADP could be found. This result indicated that IsADP is carried by sodium ions flowing into the cell. However, the dependence of IsADP on extracellular sodium was less pronounced than expected for a pure sodium current. We interpret these results to indicate that the sADP is most likely mediated by nonselective cation channels. Examination of the current underlying the sADP at different voltages indicated that this current was also voltage dependent, turning off with hyperpolarization. We conclude that the sADP elicited by muscarinic receptor activation in rat cortex is mediated predominantly by a calcium- and voltage-sensitive nonselective cation current. This current could represent an important mechanism through which ACh can regulate neuronal excitability in prefrontal cortex. PMID- 9744934 TI - Functional and ionic properties of a slow afterhyperpolarization in ferret perigeniculate neurons in vitro. AB - Intracellular recordings from spontaneously spindling GABAergic neurons of the ferret perigeniculate nucleus in vitro revealed a fast afterhyperpolarization after each action potential, a medium-duration afterhyperpolarization after each low-threshold Ca2+ spike, and a slow afterhyperpolarization after the cessation of spindle waves. The slow afterhyperpolarization was associated with an increase in membrane conductance, and the reversal potential was sensitive to extracellular [K+]o, indicating that it is mediated at least in part by the activation of a K+ conductance. However, the block of Ca2+ channels did not block the slow afterhyperpolarization, whereas the block of Na+ channels did block this event, even after the generation of repetitive Ca2+ spikes, indicating that it is mediated by a Na+-activated K+ current. Application of apamin reduced the afterhyperpolarization and enhanced a plateau potential after each low-threshold Ca2+ spike. This plateau potential could result in a prolonged depolarization of perigeniculate neurons, even before the application of apamin, resulting in the generation of tonic discharge. The plateau potential was blocked by the local application of tetrodotoxin, indicating that it is mediated by a persistent Na+ current. The activation and interaction of these slowly developing and persistent currents contributes significantly to low-frequency components of spindle wave generation. In particular, we suggest that the activation of the slow afterhyperpolarization may contribute to the generation of the spindle wave refractory period in vitro. PMID- 9744935 TI - Serum from diabetic BB/W rats enhances calcium currents in primary sensory neurons. AB - We examined the hypothesis that exposure of nondiabetic rat dorsal root ganglion (DRG) neurons to sera from diabetic BB/W rats would produce an increase in calcium currents associated with impaired regulation of the inhibitory G protein calcium channel complex. Acutely dissociated rat DRGs were incubated for 18-24 h in medium supplemented with sera (10% vol/vol) from either diabetic rats with neuropathy or age-matched, nondiabetic controls. Exposure of DRG neurons to sera from diabetic BB/W rats resulted in a surface membrane immunofluorescence pattern when treated with an anti-rat light-chain antibody that was not observed in neurons exposed to control sera. Calcium current density (IDCa) was assessed with the use of the whole cell variation of the patch-clamp technique. IDCa in neurons exposed to diabetic sera was significantly increased compared with neurons exposed to control sera. Guanine nucleotide-binding (G) protein regulation of calcium channel function was examined with the use of a two-pulse "facilitation" or IDCa enhancement protocol in the presence of activators [guanosine 5'-O-(3 thiotriphosphate) (GTP gamma S)] or antagonists [guanosine 5'-O-(2 thiodiphosphate) (GDP beta S) and pertussis toxin (PTX)] of G protein function. Facilitation was significantly decreased in neurons exposed to diabetic sera. Intracellular diffusion of neurons with GDP beta s blocked facilitation, whereas dialysis with GTP gamma s increased facilitation to a similar magnitude in neurons exposed to either diabetic or control sera. Treatment with PTX resulted in a significant increase in IDCa and approximately 50% decrease in facilitation in neurons treated with control sera but no significant changes in neurons exposed to diabetic sera. We conclude that serum from diabetic BB/W rats with neuropathy contains an autoimmune immunoglobulin that impairs regulation of the inhibitory G protein-calcium channel complex, resulting in enhanced calcium influx. Regulation of the inhibitory G protein-calcium channel complex involves PTX-sensitive and -insensitive G proteins. PMID- 9744936 TI - Recovery of locomotion after ventral and ventrolateral spinal lesions in the cat. I. Deficits and adaptive mechanisms. AB - The recovery of treadmill locomotion of eight adult cats, subjected to chronic ventral and ventrolateral spinal lesions at low thoracic levels (T11 or T13), preserving at least one dorsolateral funiculus and the dorsal columns, was documented daily using electromyographic (EMG) and kinematic methods. The data show that all cats eventually recovered quadrupedal voluntary locomotion despite extensive damage to important pathways (such as the reticulospinal and the vestibulospinal) as verified by injection of wheat germ agglutinin-horseradish peroxidase (WGA-HRP) caudal to the site of lesion. Initially (in the early period after the spinal lesion), all the cats suffered from pronounced locomotor and postural deficits, and they could not support their hindquarters or walk with their hindlimbs. Gradually, during the recovery period, they regained quadrupedal walking, although their locomotion was wobbly and inconsistent, and they suffered from poor lateral stability. EMG and kinematic data analyses showed a tendency for an increase in the variability of the step cycle duration but no major changes in the step cycle structure or in the intralimb coupling of the joints. However, the homolateral fore- and hindlimb coupling was highly perturbed in cats with the largest lesions. Although the general alternating pattern of extensor and flexors was maintained, there were various changes in the duration and amplitude of the EMG bursts as well as a lack of amplitude modulation during walking uphill or downhill on the treadmill. In cats with larger lesions, the forelimbs also seem to take a greater propulsive role than usual as revealed by a consistent increase of the activity of the triceps. In cats with smaller lesions, these deficits were transient, but, for the most extensively lesioned cats, they were pronounced and lasted long term postlesion even after reaching a more or less stable locomotor behavior (plateau period). It is concluded that recovery of quadrupedal locomotion is possible even after a massive lesion to ventral and ventrolateral quadrants, severing the vestibulospinal pathway and causing severe, although incomplete, damage to the reticulospinal tract. The quick recovery in the less lesioned cats can be attributed to remaining pathways normally implicated in locomotor function. However, in the most extensively lesioned cats, the long period of recovery and the pronounced deficits during the plateau period may indicate that the compensation, attributed to remaining reticulospinal pathways, is not sufficient and that other pathways in the dorsolateral funiculi, such as the corticospinal, can sustain and adapt, up to a certain extent, the voluntary quadrupedal walking. PMID- 9744937 TI - Mechanisms of afterhyperpolarization in lobster olfactory receptor neurons. AB - In lobster olfactory receptor neurons (ORNs), depolarizing responses to odorants and current injection are accompanied by the development of an afterhyperpolarization (AHP) that likely contributes to spike-frequency adaptation and that persists for several seconds after termination of the response. A portion of the AHP can be blocked by extracellular application of 5 mM CsCl. At this concentration, CsCl specifically blocks the hyperpolarization activated cation current (Ih) in lobster ORNs. This current is likely to be active at rest, where it provides a constant, depolarizing influence. Further depolarization deactivates Ih, thus allowing the cell to be briefly hyperpolarized when that depolarizing influence is removed, thus generating an AHP. Reactivation of Ih would terminate the AHP. The component of the AHP that could not be blocked by Cs+ (the Cs(+)-insensitive AHP) was accompanied by decreased input resistance, suggesting that this component is generated by increased conductance to an ion with an equilibrium potential more negative than the resting potential. The Cs(+)-insensitive AHP in current clamp and the underlying current in voltage clamp displayed a reversal potential of approximately -75 mV. Both EK and ECl are predicted to be in this range. Similar results were obtained with the use of a high Cl- pipette solution, although that shifted ECl from -72 mV to -13 mV. However, when EK was shifted to more positive or negative values, the reversal potential also shifted accordingly. A role for the Ca(2+)-mediated K+ current in generating the Cs(+)-independent AHP was explored by testing cells in current and voltage clamp while blocking IK(Ca) with Cs+/Co(2+)-saline. In some cells, the Cs(+)-independent AHP and its underlying current could be completely and reversibly blocked by Cs+/Co2+ saline, whereas in other cells some fraction of it remained. This indicates that the Cs(+) independent AHP results from two K+ currents, one that requires an influx of extracellular Ca2+ and one that does not. Collectively, these findings indicate that AHPs result from three phenomena that occur when lobster ORNs are depolarized: 1) inactivation of the hyperpolarization-activated cation current, 2) activation of a Ca(2+)-mediated K+ current, and 3) activation of a K+ current that does not require influx of extracellular Ca2+. Roles of these processes in modulating the output of lobster ORNs are discussed. PMID- 9744938 TI - Orphanin FQ/nociceptin inhibits synaptic transmission and long-term potentiation in rat dentate gyrus through postsynaptic mechanisms. AB - Orphanin FQ/nociceptin (OFQ), a recently characterized natural ligand for the opioid receptor-like 1 (ORL1) receptor, shares structural similarity to the endogenous opioids. Our previous study found that OFQ, like classical opioids, modulated synaptic transmission and long-term potentiation (LTP) in the hippocampal CA1 region, suggesting a modulatory role for OFQ in synaptic plasticity involved in learning and memory. In the present study we investigated the action of OFQ in the dentate gyrus and explored possible underlying cellular mechanisms. Field potential recordings showed that OFQ significantly inhibited excitatory synaptic transmission and LTP induction in the dentate lateral perforant path. In the presence of OFQ, the excitatory postsynaptic potential (EPSP) slope-population spike (E-S) curve was shifted to the right, and no significant change was found in paired-pulse facilitation, suggesting a postsynaptic mechanism responsible for the inhibition of synaptic transmission. Under whole cell voltage-clamp conditions, bath application of OFQ activated K+ currents in most granule cells tested at a holding potential of -50 mV, suggesting that OFQ could reduce the excitability of dentate granule cells by hyperpolarizing cell membranes. OFQ also inhibited the amplitude of N-methyl-D aspartate (NMDA) receptor-mediated excitatory postsynaptic currents (EPSCs) without affecting alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor-mediated EPSCs. This inhibition was not blocked by opioid receptor antagonists. Furthermore, the inward currents evoked by focal application of NMDA to granule cells were suppressed by OFQ in a dose-dependent manner, suggesting that OFQ may suppress LTP by inhibiting the function of postsynaptic NMDA receptors. These results demonstrate that OFQ may negatively modulate synaptic transmission and plasticity in the dentate gyrus through postsynaptic mechanisms, including hyperpolarization of granule cells as well as inhibition of the function of postsynaptic NMDA receptors/channels in dentate granule cells. PMID- 9744939 TI - Physiological studies of the precedence effect in the inferior colliculus of the cat. I. Correlates of psychophysics. AB - The precedence effect (PE) is experienced when two spatially separated sounds are presented with such a brief delay that only a single auditory image at or toward the location of the leading source is perceived. The responses of neurons in the central nucleus of the inferior colliculus (ICC) of cats were studied using stimuli that are known to elicit the PE, focusing on the effects of changes in stimulus conditions that a listener might encounter in a natural situation. Experiments were conducted under both free-field (anechoic chamber) and dichotic (headphones) conditions. In free field, the PE was simulated by presenting two sounds from different loudspeakers with one sound delayed relative to the other. Either click or noise stimuli (2- to 10-ms duration) were used. Dichotically, the same conditions were simulated by presenting two click or noise pairs separated by an interstimulus delay (ISD) with interaural time differences (ITDs) imposed separately for each pair. At long ISDs, all neurons responded to both leading and lagging sources as if they were delivered alone. As the ISDs were shortened, the lagging response became suppressed. The ISD of half-maximal suppression varied considerably within the population of neurons studied, ranging from 2 to 100 ms, with means of 35 and 38 ms for free field and dichotic conditions, respectively. Several correlates of psychophysical findings were observed in ICC neurons: suppression was usually stronger with lower overall stimulus level and longer duration stimuli. Suppression also was compared along the azimuth and elevation in free field by placing the lagging source at (0 degree,0 degree), which is common to both axes, and the leading sources at locations along either plane that generated similar discharge rates. All neurons that showed suppression along the azimuth also did so in the elevation. In addition, there was a high correlation in the ISD of half-maximal suppression along the two planes (r = 0.87). These findings suggest that interaural difference cues, which are robust along the horizontal axis but minimal in the median plane, are not necessary for neural correlates of the PE to be manifested. Finally, single-neuron responses did not demonstrate a correlate of build-up of suppression, a phenomenon whereby echo suppression accumulates with ongoing stimulation. This finding adds credibility to theories about the PE that argue for a "higher order" component of the PE. PMID- 9744940 TI - Physiological studies of the precedence effect in the inferior colliculus of the cat. II. Neural mechanisms. AB - We studied the responses of neurons in the inferior colliculus (IC) of cats to stimuli known to evoke the precedence effect (PE). This paper focuses on stimulus conditions that probe the neural mechanisms underlying the PE but that are not usually encountered in a natural situation. Experiments were conducted under both free-field (anechoic chamber) and dichotic (headphones) conditions. We found that in free field the amount of suppression of the lagging response depended on the location of the leading source. With stimuli in the azimuthal plane, the majority (84%) of units showed stronger suppression of the lagging response for a leading stimulus placed in the cell's responsive area as compared with a lead in the unresponsive field. A smaller number of units showed stronger suppression for a lead placed in the unresponsive field, and a few showed little effect of the lead location. In the elevational plane, there was less sensitivity of the leading source to changes in location, but for those cells in which there was sensitivity, suppression was always stronger when the lead was in the cell's responsive area. Studies on stimulus locations also were conducted under dichotic conditions by varying the interaural differences in time (ITD) of the leading source. Results were consistent with those obtained in free field, suggesting that ITDs play an important role in determining the amount of suppression that was observed as a function of leading stimulus location. In addition to location and ITD, we also studied the effect of varying the relative levels of the lead and lag as well as stimulus duration. For all units studied, increasing the level of the leading stimulus while holding the lagging stimulus constant resulted in increased suppression. Similar effects of leading source level were observed in azimuth and elevation. The effect of varying the duration of the leading source also showed that longer duration stimuli produce stronger suppression; this finding was observed both in azimuth and elevation. We also compared the suppression observed under binaural and monaural contralateral conditions and found a mixed effect: some neurons show stronger suppression under binaural conditions, others to monaural contralateral conditions, and still others show no effect. The results presented here support the hypothesis that the PE reflects a long-lasting inhibition evoked by the leading stimulus. Five possible sources for the inhibition are considered: the auditory nerve, intrinsic circuits in the cochlear nucleus, medial and lateral nuclei of the trapezoid body inhibition to the medial superior olive, dorsal nucleus of the lateral lemniscus (DNLL) inhibition to the ICC, and intrinsic circuits in the ICC itself. PMID- 9744941 TI - Changes in membrane and synaptic properties of thalamocortical circuitry caused by hydrogen peroxide. AB - Free radical (FR) production was linked to the generation of epileptiform activity. We performed electrophysiological recordings in rat thalamocortical slices to investigate the effects of FRs on the intrinsic and synaptic properties of thalamic and cortical neurons. Whole cell recordings from identified cortical pyramidal neurons and thalamic neurons of the ventrobasal nucleus revealed that exposure to the FR-forming agent H2O2 (2.5 mM) decreased gamma-aminobutyric acid A- and gamma-aminobutyric acid-B-mediated inhibition to 35.3 +/- 13.4% and 13.7 +/- 4.4% (means +/- SE) of control in cortical neurons and to 41.8 +/- 14.8% and 33.6 +/- 11.6% of control in thalamic neurons. H2O2 application increased excitatory transmission in thalamic neurons to 162.9 +/- 29.6% of control but caused no change in cortical neurons. H2O2 altered significantly the characteristic low-pass filter behavior of cortical and thalamic cells as determined by their input impedances. After 35 min of superfusion, the impedance of cortical neurons decreased by 67.0 +/- 14.5%, and thalamic decreased by 76.3 +/- 2.7% for the frequencies in the range 1-50 Hz while remaining constant for frequencies > 200 Hz. Neuronal hyperexcitability was manifested during H2O2 exposure by continuous firing and long depolarizing shifts in response to extracellular stimulation in both thalamocortical and cortical neurons only in slices preserving thalamocortical connections. In slices with severed thalamocortical connections, cortical neurons did not show signs of hyperexcitability. These observations indicate that FRs could promote hyperexcitability of thalamocortical circuits by altering the balance between excitation and inhibition and by transforming the characteristic low-pass filter behavior into a flat band-pass filter. PMID- 9744942 TI - Analysis of excitatory and inhibitory spontaneous synaptic activity in mouse retinal ganglion cells. AB - Spontaneous inhibitory and excitatory postsynaptic currents (sIPSCs and sEPSCs) were identified and characterized with whole cell and perforated patch voltage clamp recordings in adult mouse retinal ganglion cells. Pharmacological dissection revealed that all cells were driven by spontaneous synaptic inputs mediated by glutamate and gamma-aminobutyric acid-A (GABAA) receptors. One-half (7/14) of the cells also received glycinergic spontaneous synaptic inputs. Both GABAA and glycine receptor-mediated sIPSCs had rise times (10-90%) of < 1 ms. The decay times of the GABAA receptor-mediated sIPSCs were comparable with those of the glycine receptor-mediated sIPSCs. The average decay time constant for monoexponentially fitted sIPSCs was 63.2 +/- 74.1 ms (mean +/- SD, n = 3278). Glutamate receptor-mediated sEPSCs had an average rise time of 0.50 +/- 0.20 ms (n = 109) and an average monoexponential decay time constant of 5.9 +/- 8.6 ms (n = 2705). Slightly more than two-thirds of the spontaneous synaptic events were monoexponential (68% for sIPSCs and 76% for sEPSCs). The remainder of the events was biexponential. The amplitudes of the spontaneous synaptic events were not correlated with rise times, suggesting that the electrotonic filtering properties of the neurons and/or differences in the spatial location of synaptic inputs could not account for the difference between the decay time constants of the glutamate and GABAA/glycine receptor-mediated spontaneous synaptic events. The amplitudes of sEPSCs were similar to those recorded in tetrodotoxin (TTX), consistent with the events measured in control saline being the response to the release of a single quantum of transmitter. The range of the sIPSC amplitudes in control saline was wider than that recorded in TTX, consistent with some sIPSCs being evoked by presynaptic spikes having an average quantal size greater than one. The rates of sIPSCs and sEPSCs were determined under equivalent conditions by recording with perforated patch electrodes at potentials at which both types of event could be identified. Two groups of ganglion cell were observed; one group had an average sEPSCs/sIPSCs frequency ratio of 0.96 +/- 0.77 (n = 28) and another group had an average ratio of 6.63 +/- 0.82 (n = 7). These findings suggest that a subset of cells is driven much more strongly by excitatory synaptic inputs. We propose that this subset of cells could be OFF ganglion cells, consistent with the higher frequency of spontaneous action potentials found in OFF ganglion cells in other studies. PMID- 9744943 TI - Sensorimotor state of the contralateral leg affects ipsilateral muscle coordination of pedaling. AB - The objective of this study was to determine if independent central pattern generating elements controlling the legs in bipedal and unipedal locomotion is a viable theory for locomotor propulsion in humans. Coordinative coupling of the limbs could then be accomplished through mechanical interactions and ipsilateral feedback control rather than through central interlimb neural pathways. Pedaling was chosen as the locomotor task to study because interlimb mechanics can be significantly altered, as pedaling can be executed with the use of either one leg or two legs (cf. walking) and because the load on the limb can be well controlled. Subjects pedaled a modified bicycle ergometer in a two-legged (bilateral) and a one-legged (unilateral) pedaling condition. The loading on the leg during unilateral pedaling was designed to be identical to the loading experienced by the leg during bilateral pedaling. This loading was achieved by having a trained human "motor" pedal along with the subject and exert on the opposite crank the torque that the subject's contralateral leg generated in bilateral pedaling. The human "motor" was successful at reproducing each subject's one-leg crank torque. The shape of the motor's torque trajectory was similar to that of subjects, and the amount of work done during extension and flexion was not significantly different. Thus the same muscle coordination pattern would allow subjects to pedal successfully in both the bilateral and unilateral conditions, and the afferent signals from the pedaling leg could be the same for both conditions. Although the overall work done by each leg did not change, an 86% decrease in retarding (negative) crank torque during limb flexion was measured in all 11 subjects during the unilateral condition. This corresponded to an increase in integrated electromyography of tibialis anterior (70%), rectus femoris (43%), and biceps femoris (59%) during flexion. Even given visual torque feedback in the unilateral condition, subjects still showed a 33% decrease in negative torque during flexion. These results are consistent with the existence of an inhibitory pathway from elements controlling extension onto contralateral flexion elements, with the pathway operating during two-legged pedaling but not during one-legged pedaling, in which case flexor activity increases. However, this centrally mediated coupling can be overcome with practice, as the human "motor" was able to effectively match the bilateral crank torque after a longer practice regimen. We conclude that the sensorimotor control of a unipedal task is affected by interlimb neural pathways. Thus a task performed unilaterally is not performed with the same muscle coordination utilized in a bipedal condition, even if such coordination would be equally effective in the execution of the unilateral task. PMID- 9744945 TI - Amiloride blocks acid responses in NaCl-best gustatory neurons of the hamster solitary nucleus. AB - Biophysical studies of isolated taste receptor cells show that one mechanism of Na+ salt transduction involves the inward movement of Na+ through amiloride blockable ion channels on the apical receptor cell membrane, which leads to a direct depolarization. Hamster taste receptor cells with amiloride-blockable Na+ responses also show an amiloride-sensitive H+ current. Thus one mechanism for the transduction of acid taste involves the amiloride-sensitive channel. We investigated the effects of amiloride on responses to acids in neurons of the nucleus of the solitary tract (NST) of the hamster. The responses of 47 NST neurons were recorded extracellularly while the anterior tongue was stimulated with solutions representing the four taste qualities (NaCl, sucrose, HCl, quinine), which were used to characterize each cell on the basis of its best stimulus. The effects of amiloride on responses to 10 mM HCl, 10 mM citric acid, 100 mM NaCl, and 100 mM sucrose were then investigated. Stimuli were presented alone for 30 s (control trials) and also presented for 10 s, followed by a mixture of the stimulus with 10 microM amiloride for 10 s, followed by the stimulus alone again for 10 s (amiloride trials). The effects of amiloride were assessed by comparing the responses of cells with the stimulus + amiloride with that of the stimulus alone. In neurons classified as NaCl-best, amiloride reversibly blocked responses to NaCl, HCl, and citric acid. In HCl-best neurons, amiloride had no effect on responses to any of these stimuli. In sucrose-best neurons, amiloride blocked the response to NaCl but not to sucrose or to either acid. These results support the hypothesis that acids are transduced by at least two different receptor mechanisms in the hamster, amiloride sensitive and amiloride insensitive. At the NST, these inputs are tightly maintained in two separate populations of neurons. Sucrose-best neurons, which show amiloride effects on NaCl but not acids, appear to receive converging inputs from both amiloride-sensitive (N-best) and amiloride-insensitive (H-best) chorda tympani nerve fibers. PMID- 9744944 TI - Involvement of Ras/MAP kinase in the regulation of Ca2+ channels in adult bullfrog sympathetic neurons by nerve growth factor. AB - The cellular mechanisms that underlie nerve growth factor (NGF) induced increase in Ca(2+)-channel current in adult bullfrog sympathetic B-neurons were examined by whole cell recording techniques. Cells were maintained at low density in neuron-enriched, defined-medium, serum-free tissue culture for 6 days in the presence or absence of NGF (200 ng/ml). The increase in Ba2+ current (IBa) density induced by NGF was attenuated by the RNA synthesis inhibitor cordycepin (20 microM), by the DNA transcription inhibitor actinomycin D (0.01 microgram/ml), by inhibitors of Ras isoprenylation (perillic acid 0.1-1.0 mM or alpha-hydroxyfarnesylphosphonic acid 10-100 microM), by tyrosine kinase inhibitors genistein (20 microM) or lavendustin A (1 microM), and by PD98059 (10 100 microM), an inhibitor of mitogen-activated protein kinase kinase. Inhibitors of the phosphatidylinositol 3-kinase (PI3K) pathway (wortmannin, 100 nM, or LY29400, 100 microM) were ineffective as were inhibitors of phospholipase C gamma (U73122 or neomycin, both 100 microM). The effect of NGF persisted in Ca(2+)-free medium that contained 1.8 mM Mg2+ and 2 mM ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid. It was mimicked by a Trk antibody that was capable of inducing neurite outgrowth in explant cultures of bullfrog sympathetic ganglion. Antibodies raised against the low-affinity p75 neurotrophin receptor were ineffective in blocking the effect of NGF on IBa. These results suggest that NGF-induced increase in Ca2+ channel current in adult sympathetic neurons results, at least in part, from new channel synthesis after Trk activation of Ras and mitogen activated protein kinase by a mechanism that is independent of extracellular Ca2+. PMID- 9744946 TI - Hand dominance and motor unit firing behavior. AB - Daily preferential use was shown to alter physiological and mechanical properties of skeletal muscle. This study was aimed at revealing differences in the control strategy of muscle pairs in humans who show a clear preference for one hand. We compared the motor unit (MU) recruitment and firing behavior in the first dorsal interosseous (FDI) muscle of both hands in eight male volunteers whose hand preference was evaluated with the use of a standard questionnaire. Myoelectric signals were recorded while subjects isometrically abducted the index finger at 30% of the maximal voluntary contraction (MVC) force. A myoelectric signal decomposition technique was used to accurately identify MU firing times from the myoelectric signal. In MUs of the dominant hand, mean values for recruitment threshold, initial firing rate, average firing rate at target force, and discharge variability were lower when compared with the nondominant hand. Analysis of the cross-correlation between mean firing rate and muscle force revealed cross-correlation peaks of longer latency in the dominant hand than in the nondominant side. This lag of the force output with respect to fluctuations in the firing behavior of MUs is indicative of a greater mechanical delay in the dominant FDI muscle. MVC force was not significantly different across muscle pairs, but the variability of force at the submaximal target level was higher in the nondominant side. The presence of lower average firing rates, lower recruitment thresholds, and greater firing rate/force delay in the dominant hand is consistent with the notion of an increased percentage of slow twitch fibers in the preferentially used muscle, allowing twitch fusion and force buildup to occur at lower firing rates. It is suggested that a lifetime of preferred use may cause adaptations in the fiber composition of the dominant muscle such that the mechanical effectiveness of its MUs increased. PMID- 9744947 TI - Cutaneous responsiveness of lumbar spinal dorsal horn neurons is reduced by general anesthesia, an effect dependent in part on GABAA mechanisms. AB - Extracellular activity was recorded from single spinal dorsal horn neurons in both chronic cat and acute rat models. This was done to define the effects of anesthesia on the processing of sensory information elicited by nonnoxious tactile stimulation of peripheral receptive fields (RFs). In the chronic cat model, baseline data were obtained in physiologically intact, awake, drug-free animals before anesthetic administration (halothane 1.0-2.0%). This made it possible to compare and contrast activity of each cell in the drug-free and anesthetized state. Halothane effects were confirmed in the acute rat model (anesthetized, spinally transected, and in some cases decerebrate). In addition, the gamma-aminobutyic acid-A (GABAA)-receptor antagonist picrotoxin (2 mg/kg) was administered intravenously to verify that the observed halothane effect on spinal dorsal horn neurons was mediated by an interaction with GABAA-receptor systems. Halothane effects on three separate measures of response to nonnoxious tactile stimuli were observed in the chronic cat model. Halothane produced a significant, dose-dependent reduction in the low-threshold RF area of the neurons studied. Halothane also caused a significant reduction in neuronal response to RF brushing (dynamic stimulus) and to maintained contact with the RF (static stimulus). A dose dependency was not observed with these latter two effects. Neurons with a predominant rapidly adapting response seemed to be less susceptible to halothane suppression than slowly adapting cells. In the acute rat model an increase in halothane caused a reduction in neuronal response similar to that seen in the cat. The intravenous administration of 2 mg/kg of picrotoxin by itself caused no significant change in RF size or response to brushing. However, the same amount of picrotoxin did cause a 50% reversal of the halothane-induced reduction in RF size without causing a significant change in the halothane effect on response to RF brushing. In contrast to work recently reported in a chronic sheep model, halothane causes a significant reduction in spinal dorsal horn neuronal response to tactile stimulation of peripheral RFs. This effect is caused by, in part, but not exclusively, to GABAA-neurotransmitter systems. However, the relative influence of GABAA systems may vary with the nature of the stimulus. PMID- 9744948 TI - Computer simulation of post-spike facilitation in spike-triggered averages of rectified EMG. AB - When the spikes of a motor cortical cell are used to compile a spike-triggered average (STA) of rectified electromyographic (EMG) activity, a post-spike facilitation (PSF) is sometimes seen. This is generally thought to be indicative of direct corticomotoneuronal (CM) connections. However, it has been claimed that a PSF could be caused by synchronization between CM and non-CM cells. This study investigates the generation of PSF using a computer model. A population of cortical cells was simulated, some of which made CM connections to a pool of 103 motoneurons. Motoneurons were simulated using a biophysically realistic model. A subpopulation of the cortical cells was synchronized together. After a motoneuron discharge, a motor unit action potential was generated; these were summed to produce an EMG output. Realistic values were used for the corticospinal and peripheral nerve conduction velocity distribution, for slowing of impulse conduction in CM terminal axons, and for the amount of cortical synchrony. STA of the rectified EMG from all cortical neurons showed PSF; however, these were qualitatively different for CM versus non-CM cells. Using an epoch analysis to determine reliability in a quantitative manner, it was shown that the onset latency of PSF did not distinguish the two classes of cells after 10,000 spikes because of high noise in the averages. The time of the PSF peak and the peak width at half-maximum (PWHM) could separate CM from synchrony effects. However, only PWHM was robust against changes in motor unit action-potential shape and duration and against changes in the width of cortical synchrony. The amplitude of PSF from a CM cell could be doubled by the presence of synchrony. It is proposed that, if a PSF has PWHM < 7 ms, this reliably indicates that the trigger is a CM cell projecting to the muscle whose EMG is averaged. In an analysis of experimental data where macaque motor cortical cells facilitated hand and forearm muscle EMG, 74% of PSFs fulfilled this criterion. The PWHM criterion could be applied to other STA studies in which it is important to exclude the effects of synchrony. PMID- 9744949 TI - Dependence on target configuration of express saccade-related activity in the primate superior colliculus. AB - To help understand how complex visual stimuli are processed into short-latency saccade motor programs, the activity of visuomotor neurons in the deeper layers of the superior colliculus was recorded while two monkeys made express saccades to one target and to two targets. It has been shown previously that the visual response and perimotor discharge characteristic of visuomotor neurons temporally coalesce into a single burst of discharge for express saccades. Here we seek to determine whether the distributed visual response to two targets spatially coalesces into a command appropriate for the resulting saccade. Two targets were presented at identical radial eccentricities separated in direction by 45 degrees. A gap paradigm was used to elicit express saccades. Express saccades were more likely to land in between the two targets than were saccades of longer latency. The speeds of express saccades to two targets were similar to those of one target of similar vector, as were the trajectories of saccades to one and two targets. The movement fields for express saccades to two targets were more broad than those for saccades to one target for all neurons studied. For most neurons, the spatial pattern of discharge for saccades to two targets was better explained as a scaled version of the visual response to two spatially separate targets than as a scaled version of the perimotor response accompanying a saccade to a single target. Only the discharge of neurons with large movement fields could be equally well explained as a visual response to two targets or as a perimotor response for a one-target saccade. For most neurons, the spatial properties of discharge depended on the number of targets throughout the entire saccade-related burst. These results suggest that for express saccades to two targets the computation of saccade vector is not complete at the level of the superior colliculus for most neurons and an explicit process of target selection is not necessary at this level for the programming of an express saccade. PMID- 9744950 TI - Resonance effect for neural spike time reliability. AB - The spike timing reliability of Aplysia motoneurons stimulated by repeated presentation of periodic or aperiodic input currents is investigated. Two properties of the input are varied, the frequency content and the relative amplitude of the fluctuations to the mean (expressed as the coefficient of variation: CV). It is shown that, for small relative amplitude fluctuations (CV approximately 0.05-0.15), the reliability of spike timing is enhanced if the input contains a resonant frequency equal to the firing rate of the neuron in response to the DC component of the input. This resonance-related enhancement in reliability decreases as the relative amplitude of the fluctuations increases (CV ->1). Similar results were obtained for a leaky integrate-and-fire neuronal model, suggesting that these effects are a general property of encoders that combine a threshold with a leaky integrator. These observations suggest that, when the magnitude of input fluctuations is small, changes in the power spectrum of the current fluctuations or in the spike discharge rate can have a pronounced effect on the ability of the neuron to encode a time-varying input with reliably timed spikes. PMID- 9744951 TI - Spike-wave complexes and fast components of cortically generated seizures. I. Role of neocortex and thalamus. AB - We explored the relative contributions of cortical and thalamic neuronal networks in the generation of electrical seizures that include spike-wave (SW) and polyspike-wave (PSW) complexes. Seizures were induced by systemic or local cortical injections of bicuculline, a gamma-aminobutyric acid-A (GABAA) antagonist, in cats under barbiturate anesthesia. Field potentials and extracellular neuronal discharges were recorded through arrays of eight tungsten electrodes (0.4 or 1 mm apart) placed over the cortical suprasylvian gyrus and within the thalamus. 1) Systemic injections of bicuculline induced SW/PSW seizures in cortex, whereas spindle sequences continued to be present in the thalamus. 2) Cortical suprasylvian injection of bicuculline induced focal paroxysmal single spikes that developed into full-blown seizures throughout the suprasylvian cortex. The seizures were characterized by highly synchronized SW or PSW complexes at 2-4 Hz, interspersed with runs of fast (10-15 Hz) activity. The intracellular aspects of this complex pattern in different types of neocortical neurons are described in the following paper. Complete decortication abolished the seizure, leaving intact thalamic spindles. Injections of bicuculline in the cortex of athalamic cats resulted in similar components as those occurring with an intact thalamus. 3) Injection of bicuculline in the thalamus decreased the frequency of barbiturate spindles and increased the synchrony of spike bursts fired by thalamocortical and thalamic reticular cells but did not induce seizures. Decortication did not modify the effects of bicuculline injection in the thalamus. Our results indicate that the minimal substrate that is necessary for the production of seizures consisting of SW/PSW complexes and runs of fast activity is the neocortex. PMID- 9744952 TI - Spike-wave complexes and fast components of cortically generated seizures. II. Extra- and intracellular patterns. AB - In the previous paper we have demonstrated, by means of field potential and extracellular unit recordings, that bicuculline-induced seizures, which include spike-wave (SW) or polyspike-wave (PSW) complexes, are initiated intracortically and survive ipsilateral thalamectomy. Here, we used multisite field potential and extracellular recordings to validate the patterns of cortical SW/PSW seizures in chronically implanted, behaving cats. To investigate the cellular patterns and excitability during spontaneously occurring and electrically elicited cortical seizures, we used single and dual intracellular recordings from regular-spiking (RS) and fast-rhythmic-bursting (FRB) cortical neurons, in conjunction with field potential recordings from neocortex and related thalamic nuclei, in cats maintained under ketamine-xylazine anesthesia. 1) Invariably, the spontaneous or electrically induced seizures were initiated within the cortex of both behaving and anesthetized animals. Spontaneously occurring, compound seizures consisting of SW/PSW complexes at 2-4 Hz and fast runs at 10-15 Hz, developed without discontinuity from the slow (mainly 0.5-0.9 Hz), sleeplike, cortically generated oscillation. 2) During SW/PSW complexes, RS neurons discharged spike trains during the depth-negative component of the cortical "spike" component of field potentials and were hyperpolarized during the depth-positive field wave. The FRB neurons fired many more action potentials than RS cells during SW/PSW complexes. Averaged activities triggered by the spiky field potentials or by the steepest slope of depolarization in cortical neurons demonstrated similar relations between intracellular activities and field potentials during sleep and seizure epochs, the latter-being an exaggeration of the depolarizing and hyperpolarizing components of the slow sleep oscillation. 3) During the fast runs, RS cells were tonically depolarized and discharged single action potentials or spike doublets (usually with pronounced spike inactivation), whereas FRB cells discharged rhythmic spike bursts, time locked with the depth-negative field potentials. 4) Neuronal excitability, tested by depolarizing current pulses applied throughout the seizures and compared with pre- and postseizure epochs, showed a decreased number of evoked action potentials during both seizure components (SW/PSW complexes and fast runs), eventually leading to null responses during the postictal depression. 5) Data suggest that interconnected FRB neurons may play an important role in the initiation of cortical seizures. We discuss the similarities between the electrographic patterns described in this study and those found in different forms of clinical seizures. PMID- 9744953 TI - Spike-wave complexes and fast components of cortically generated seizures. III. Synchronizing mechanisms. AB - The intracortical and thalamocortical synchronization of spontaneously occurring or bicuculline-induced seizures, consisting of spike-wave (SW) or polyspike-wave (PSW) complexes at 2-3 Hz and fast runs at 10-15 Hz, was investigated in cats under ketamine-xylazine anesthesia. We used single and dual simultaneous intracellular recordings from cortical areas 5 and 7, and extracellular recordings of unit firing and field potentials from neocortical areas 5, 7, 17, 18, as well as related thalamic nuclei. The evolution of time delays between paroxysmal depolarizing events in single neurons or neuronal pools recorded from adjacent and distant sites was analyzed by using 1) sequential cross-correlations between field potentials, 2) averaged activities triggered by the spiky component of cortical SW/PSW complexes, and 3) time histograms between neuronal discharges. In all instances, the paroxysmal activities recorded from the dorsal thalamus lagged the onset of seizures in neocortex. The time lags between simultaneously impaled cortical neurons were significantly smaller during SW complexes than during the prior epochs of slow oscillation. During seizures, as during the slow oscillation, the intracortical synchrony was reduced with increased distance between different cortical sites. Dual intracellular recordings showed that, during the same seizure, time lags were not constant and, instead, reflected alternating precession of the recorded foci. After transection between areas 5 and 7, the intracortical synchrony was lost, but corticothalamocortical volleys could partially restore seizure synchrony. These data show that the neocortex leads the thalamus during SW/PSW seizures, that time lags between cortical foci are not static, and that thalamus may assist synchronization of SW/PSW seizures after disconnection of intracortical synaptic linkages. PMID- 9744954 TI - Spike-wave complexes and fast components of cortically generated seizures. IV. Paroxysmal fast runs in cortical and thalamic neurons. AB - In the preceding papers of this series, we have analyzed the cellular patterns and synchronization of neocortical seizures occurring spontaneously or induced by electrical stimulation or cortical infusion of bicuculline under a variety of experimental conditions, including natural states of vigilance in behaving animals and acute preparations under different anesthetics. The seizures consisted of two distinct components: spike-wave (SW) or polyspike-wave (PSW) at 2-3 Hz and fast runs at 10-15 Hz. Because the thalamus is an input source and target of cortical neurons, we investigated here the seizure behavior of thalamic reticular (RE) and thalamocortical (TC) neurons, two major cellular classes that have often been implicated in the generation of paroxysmal episodes. We performed single and dual simultaneous intracellular recordings, in conjunction with multisite field potential and extracellular unit recordings, from neocortical areas and RE and/or dorsal thalamic nuclei under ketamine-xylazine and barbiturate anesthesia. Both components of seizures were analyzed, but emphasis was placed on the fast runs because of their recent investigation at the cellular level. 1) The fast runs occurred at slightly different frequencies and, therefore, were asynchronous in various cortical neuronal pools. Consequently, dorsal thalamic nuclei, although receiving convergent inputs from different neocortical areas involved in seizure, did not express strongly synchronized fast runs. 2) Both RE and TC cells were hyperpolarized during seizure episodes with SW/PSW complexes and relatively depolarized during the fast runs. As known, hyperpolarization of thalamic neurons deinactivates a low-threshold conductance that generates high-frequency spike bursts. Accordingly, RE neurons discharged prolonged high-frequency spike bursts in close time relation with the spiky component of cortical SW/PSW complexes, whereas they fired single action potentials, spike doublets, or triplets during the fast runs. In TC cells, the cortical fast runs were reflected as excitatory postsynaptic potentials appearing after short latencies that were compatible with monosynaptic activation through corticothalamic pathways. 3) The above data suggested the cortical origin of these seizures. To further test this hypothesis, we performed experiments on completely isolated cortical slabs from suprasylvian areas 5 or 7 and demonstrated that electrical stimulation within the slab induces seizures with fast runs and SW/PSW complexes, virtually identical to those elicited in intact brain animals. The conclusion of all papers in this series is that complex seizure patterns, resembling those described at the electroencephalogram level in different forms of clinical seizures with SW/PSW complexes and, particularly, in the Lennox-Gastaut syndrome of humans, are generated in neocortex. Thalamic neurons reflect cortical events as a function of membrane potential in RE/TC cells and degree of synchronization in cortical neuronal networks. PMID- 9744955 TI - Similar propagation of SD and hypoxic SD-like depolarization in rat hippocampus recorded optically and electrically. AB - Neuron membrane changes and ion redistribution during normoxic spreading depression (SD) induced, for example, by potassium injection, closely resemble those that occur during hypoxic SD-like depolarization (HSD) induced by oxygen withdrawal, but the degree to which the two phenomena are related is controversial. We used extracellular electrical recording and imaging of intrinsic optical signals in hippocampal tissue slices to compare 1) initiation and spread of these two phenomena and 2) the effects of putative gap junction blocking agents, heptanol and octanol. Both events arose focally, after which a clear advancing wave front of increased reflectance and DC shift spread along the CA1 stratum radiatum and s. oriens. The rate of spread was similar: conduction velocity of normoxic SD was 8.73 +/- 0.92 mm/min (mean +/- SE) measured electrically and 5.84 +/- 0.63 mm/min measured optically, whereas HSD showed values of 7.22 +/- 1.60 mm/min (electrical) and 6.79 +/- 0.42 mm/min (optical). When initiated in CA1, normoxic SD consistently failed to enter the CA3 region (7/7 slices) and could not be initiated by direct KC1 injection in the CA3 region (n = 3). Likewise, the hypoxic SD-like optical signal showed onset in the CA1 region and halted at the CA1/CA3 boundary (9/9 slices), but in some (4/9) slices the dentate gyrus region showed a separate onset of signal changes. Microinjection into CA1 stratum radiatum of octanol (1 mM), which when bath applied arrests the spread of normoxic SD, created a small focus that appeared to be protected from hypoxic depolarization. However, bath application of heptanol (3 mM) or octanol (2 mM) did not prevent the spread of HSD, although the onset was delayed. This suggests that, although gap junctions may be essential for the spread of normoxic SD, they may play a less important role in the spread of HSD. PMID- 9744956 TI - Refractory periods observed by intrinsic signal and fluorescent dye imaging. AB - All perfusion-based imaging modalities depend on the relationship between neuronal and vascular activity. However, the relationship between stimulus and response was never fully characterized. With the use of optical imaging (intrinsic signals and intravascular fluorescent dyes) during repetitive stimulation paradigms, we observed reduced responses with temporally close stimuli. Cortical evoked potentials, however, did not produce the same reduced responsiveness. We therefore termed these intervals of reduced responsiveness "refractory periods." During these refractory periods an ability to respond was retained, but at a near 60% reduction in the initial magnitude. Although increasing the initial stimulus duration lengthened the observed refractory periods, significantly novel or temporally spaced stimuli overcame them. We observed this phenomenon in both rodent and human subjects in somatosensory and auditory cortices. These results have significant implications for understanding the capacities, mechanisms, and distributions of neurovascular coupling and thereby possess relevance to all perfusion-dependent functional imaging techniques. PMID- 9744957 TI - Functional MRI study of thalamic and cortical activations evoked by cutaneous heat, cold, and tactile stimuli. AB - Positron emission tomography studies have provided evidence for the involvement of the thalamus and cortex in pain and temperature perception. However, the involvement of these structures in pain and temperature perception of individual subjects has not been studied in detail with high spatial resolution imaging. As a first step toward this goal, we have used functional magnetic resonance imaging (fMRI) to locate discrete regions of the thalamus, insula, and second somatosensory cortex (S2) modulated during innocuous and noxious thermal stimulation. Results were compared with those obtained during tactile stimulation of the palm. High resolution functional images were acquired on a 1.5 T echospeed GE MR system with an in-plane resolution of 1.7 mm. A modified peltier-type thermal stimulator was used to deliver innocuous cool and warm and noxious cold and hot stimuli for 40-60 s to the thenar eminence of normal male and female volunteers. Experimental paradigms consisted of four repetitions of interleaved control and task stimuli. A pixel by pixel statistical analysis of images obtained during each task versus control (e.g., noxious heat vs. warm, warm vs. neutral temperature, etc.) was used to determine task-related activations. Painful thermal stimuli activated discrete regions within the lateral and medial thalamus, and insula, predominantly in the anterior insula in most subjects, and the contralateral S2 in 50% of subjects. The innocuous thermal stimuli did not activate the S2 in any of the subjects but activated the thalamus and posterior insula in 50% of subjects. By comparison, innocuous tactile stimulation consistently activated S2 bilaterally and the contralateral lateral thalamus. These data also demonstrate that noxious thermal and innocuous tactile-related activations overlap in S2. The data also suggest that innocuous and noxious related activations may overlap within the thalamus but may be located in different regions of the insula. Therefore, we provide support for a role of the anterior insula, S2, and thalamus in the perception of pain; whereas the posterior insula appears to be involved in tactile and innocuous temperature perception. These data demonstrate the feasibility of using fMRI for studies of pain, temperature, and mechanical stimuli in individual subjects, even in small regions such as thalamic nuclei. However, the intersubject variability should be considered in future single subject imaging studies and studies that rely on averaged group responses. PMID- 9744958 TI - Activation of protein kinase C increases neuronal excitability by regulating persistent Na+ current in mouse neocortical slices. AB - Effects of the protein kinase C activating phorbol ester, phorbol 12-myristate 13 acetate (PMA), were studied in whole cell recordings from layer V neurons in slices of mouse somatosensory neocortex. PMA was applied intracellularly (100 nM to 1 microM) to restrict its action to the cell under study. In current-clamp recordings, it enhanced neuronal excitability by inducing a 10- to 20-mV decrease in voltage threshold for action-potential generation. Because spike threshold in neocortical neurons critically depends on the properties of persistent Na+ current (INaP), effects of PMA on this current were studied in voltage clamp. After blocking K+ and Ca2+ currents, INaP was revealed by applying slow depolarizing voltage ramps from -70 to 0 mV. Intracellular PMA induced a decrease in INaP at very depolarized membrane potentials. It also shifted activation of INaP in the hyperpolarizing direction, however, such that there was a significant increase in persistent inward current at potentials more negative than -45 mV. When tetrodotoxin (TTX) was added to the bath, blocking INaP and leaving only an outward nonspecific cationic current (Icat), PMA had no apparent effect on responses to voltage ramps. Thus PMA did not affect Icat, and it did not induce any additional current. Intracellular application of the inactive PMA analogue, 4 alpha-PMA, did not affect INaP. The specific protein kinase C inhibitors, chelerythrine (20 microM) and calphostin C (10 microM), blocked the effect of PMA on INaP. The data suggest that PMA enhances neuronal excitability via a protein kinase C-mediated increase in INaP at functionally critical subthreshold voltages. This novel effect would modulate all neuronal functions that are influenced by INaP, including synaptic integration and active backpropagation of action potential from the soma into the dendrites. PMID- 9744959 TI - Compensation for gaze perturbation during inactivation of the caudal fastigial nucleus in the head-unrestrained cat. AB - Muscimol injection in the caudal part of the fastigial nucleus (cFN) leads, in the head-unrestrained cat, to a characteristic dysmetria of saccadic gaze shifts toward visual targets. The goal of the current study was to test whether this pharmacological cFN inactivation impaired the ability to compensate for unexpected perturbations in gaze position during the latency period of the saccadic response. Such perturbations consisted of moving gaze away from the target by a transient electrical microstimulation in the deep layers of the superior colliculus simultaneously with extinction of the visual target. After injection of muscimol in the cFN, targets located in the contralesional hemifield elicited gaze shifts that fell short of the target in both "perturbed" and "unperturbed" trials. The amplitude of the compensatory contraversive gaze shifts in perturbed trials coincided with the predicted amplitude of unperturbed responses starting from the same position. Targets located in the opposite hemifield elicited hypermetric gaze shifts in both trial types, and the error of compensatory responses was not statistically different from that of unperturbed gaze shifts. These results indicate that inactivation of the cFN does not interfere with the ability of the head-unrestrained cat to compensate for ipsiversive or contraversive perturbations in gaze position. Thus the gaze related feedback signals that are used to compute a reference signal of desired gaze displacement are not impaired by cFN inactivation. PMID- 9744960 TI - Amplification of perforant-path EPSPs in CA3 pyramidal cells by LVA calcium and sodium channels. AB - The perforant path forms a monosynaptic connection between the cells of layer II of the entorhinal cortex and the pyramidal cells in hippocampal area CA3. Although this projection is prominent anatomically, very little is known about the physiological properties of this input. The distal location of these synapses suggests that somatically recorded perforant-path excitatory postsynaptic potentials (EPSPs) may be influenced by the activation of voltage-dependent channels in CA3 cells. We observed that perforant-path EPSPs are reduced (by approximately 25%) by blockade of postsynaptic low-voltage-activated calcium and sodium channels, indicating that perforant-path EPSPs are amplified by the activation of these channels. These data suggest that the perforant path may represent an important and highly modifiable direct connection between the entorhinal cortex and area CA3. PMID- 9744961 TI - Movement sequence-related activity reflecting numerical order of components in supplementary and presupplementary motor areas. AB - The supplementary motor area (SMA) and presupplementary motor areas (pre-SMA) have been implicated in movement sequencing, and neurons in SMA have been shown to encode what might be termed the relational order among sequence components (e.g., movement X followed by movement Y). To determine whether other aspects of movement sequencing might also be encoded by SMA or pre-SMA neurons, we analyzed task-related activity recorded from both areas in conjunction with a sequencing task that dissociated the numerical order of components (e.g., movement X as the 2nd component, irrespective of which movements precede or follow X). Sequences were constructed from eight component movements, each characterized by three spatial variables (origin, direction, and endpoint). Task-related activity recorded from 56 SMA and 63 pre-SMA neurons was categorized according to both the epoch (delay, reaction time, and movement time) and the spatial variable or component movement with which it was associated. All but one instance of task related activity was selective for one of the spatial variables (SV-selective) rather than for any of the component movements themselves. Of 110 instances of SV selective activity in SMA, 43 (39%) showed significant effects of numerical order. The corresponding incidence in pre-SMA, 82 (71%) of 116, was substantially higher (P < 0.00001). No effects of numerical order were evident among the hand paths, movement times, or electromyographic activity associated with task performance. We concluded that neurons in SMA and pre-SMA may encode the numerical order of components, at least for sequences that are distinguished mainly by that aspect of component ordering. PMID- 9744962 TI - Alterations in the balance of protein kinase/phosphatase activities parallel reduced synaptic strength during aging. AB - The current research examined the regulation of synaptic strength by protein phosphorylation during aging. Bath application of the protein phosphatase 1 and 2A (PP1 and PP2A) inhibitor calyculin A (1 microM) enhanced CA3-CA1 synaptic strength in hippocampal slices from aged male (20-24 mo) but not from young adult male (4-6 mo) Fischer 344 rats. Similarly, injection of the PP1 and PP2A inhibitor microcystin-L,R (5 microM) into CA1 cells caused an increase in the intracellular synaptic response only in slices from aged rats. In contrast, bath application of the serine/threonine kinase inhibitor H-7 (10 microM) induced a decrease in synaptic strength only in slices from the young adult group. These results demonstrate that phosphorylation dependent regulation of intrinsic synaptic efficacy changes during aging. PMID- 9744963 TI - GABAB receptor-mediated modulation of presynaptic currents and excitatory transmission at a fast central synapse. AB - Large nerve terminals (calyces of Held) in the medial nucleus of the trapezoid body (MNTB) offer a unique opportunity to explore the modulation of presynaptic channels at a mammalian central synapse. In this study I examined gamma aminobutyric acid-B (GABAB)-mediated presynaptic inhibition at the calyx of Held in slices of the rat auditory brain stem. The selective GABAB agonist baclofen caused a potent inhibition of synaptic transmission and presynaptic Ca2+ current. The inhibition of presynaptic Ca2+ channels was associated with a slowing of the activation kinetics of the underlying current, and the inhibition was relieved by strong depolarization. The inhibition of both synaptic transmission and presynaptic Ca2+ current was abolished by N-ethylmaleimide, a sulfhydryl alkylating agent that uncouples the G(o)/Gi class of G proteins from receptors. Baclofen does not activate a potassium conductance in the presynaptic terminal. Taken together, these results suggest that GABAB receptors inhibit synaptic transmission via G protein-mediated modulation of presynaptic Ca2+ channels at this large central synapse. Furthermore, these findings demonstrate that basic mechanisms of G protein-mediated inhibition of Ca2+ channels, proposed from recordings of neuron cell bodies, are well conserved at nerve endings in the mammalian brain. PMID- 9744964 TI - Changes in the temporal pattern of primary motor cortex activity in a directional isometric force versus limb movement task. AB - We recorded the activity of 75 proximal-arm-related cells in caudal primary motor cortex (MI) while a monkey generated either isometric forces or limb movements against an inertial load. The forces and movements were in eight directions in a horizontal plane. The isometric force generated at the hand increased monotonically in the direction of the target force level. The force exerted against the load in the movement task was more complex, including a transient decelerative phase during the movement as the hand approached the target. Electromyographic (EMG) activity of proximal-arm muscles reflected the task dependent changes in dynamics, showing a ramp increase in activity during the isometric task and a reciprocal triphasic burst pattern in the movement task. A sliding 50-ms window analysis showed that the directionality of the EMG, when expressed in hand-centered spatial coordinates, remained stable throughout the isometric ramp but often showed a significant transient shift during the limb movements. Many cells in M1 showed corresponding significant changes in activity pattern and instantaneous directionality between the two tasks. This momentary dissociation of discharge from the directional kinematics of hand displacement is evidence that the activity of many single proximal-arm related M1 cells is not coupled only to the direction and velocity of hand motion. PMID- 9744965 TI - Reflex suppression in the anti-saccade task is dependent on prestimulus neural processes. AB - Reflexive responses often must be suppressed to correctly execute a voluntary behavior. It is largely unknown why this control sometimes fails. To examine the neural processes responsible for these failures, we recorded single-neuron activity in the superior colliculus (SC) in behaving monkeys during an anti saccade task in which they had to suppress a saccade to a visual stimulus that suddenly appeared in the periphery and generate a saccade to the opposite side. We found that the level and distribution of prestimulus activity of buildup neurons in the SC was highly predictive of whether a correct response or an error occurred. A high level of prestimulus activity in buildup neurons at the location in the SC where the visual stimulus was represented was associated with the generation of a reflexive saccade to the stimulus. These findings suggest that the successful suppression of reflexive saccades is dependent on prestimulus neural processes in the SC. PMID- 9744966 TI - Time-dependent motor memory processes in amnesic subjects. AB - Functional properties of motor memory change with the passage of time. The time dependent nature of memories in humans has also been demonstrated for certain "declarative" memories. When the declarative memory system is damaged, are the time-dependent properties associated with motor memories intact? To approach this question, we examined five subjects with global amnesia (AMN), including subject H.M., and a group of age-matched control subjects. The task was to make reaching movements to visually presented targets. We found that H.M. (but not the other subjects) was significantly impaired in the ability to perform the visuomotor kinematic transformations required in this task, to accurately move the hand in the direction specified by a target. With extensive practice, H.M.'s performance improved significantly. At this point, a force field was imposed on the hand. With practice in field A, H.M. and other AMN subjects developed aftereffects and maintained these aftereffects for 24 h. To quantify postpractice properties associated with motor memories, subjects learned field B on day 2 and at 5 min were retested in field A. In both subject groups, performance in field A was significantly worse than their own naive performance a day earlier. The aftereffects indicated persistence of the just-learned but now inappropriate motor memory. After 4 h of rest, subjects were retested in B. Performance was now at naive levels. The aftereffects at 4 h indicated a reduced influence of the memory of field A. The time-dependent patterns of motor memory perseveration, as measured at 5 min and 4 h, were not different in the AMN and normal control groups. PMID- 9744967 TI - Oscillatory activity in the cerebellar hemispheres of unrestrained rats. AB - We recorded multiunit neural activity in the granule cell layer of cerebellar folium Crus IIa in unrestrained rats. Seven- to 8-Hz oscillatory activity was seen during behavioral states in which the animal was immobile; any movement the animal made coincided with termination of the oscillations. However, nearly one third of oscillatory episodes appeared to cease spontaneously, in the absence of any observable sensory input or movement. Oscillations were synchronized both within and between cerebellar hemispheres, demonstrating precise temporal coordination among multiple, bilateral levels of the somatosensory system. We interpret these data in the context of similar oscillations observed in other brain structures and suggest that the oscillations are an underlying dynamic property of the entire somatosensory network. PMID- 9744968 TI - Efference copy provides the eye position information required for visually guided reaching. AB - The contribution of extraocular muscle (EOM) proprioception to the eye position signal used to transform retinotopic visual information to a craniotopic reference frame remains uncertain. In this study we examined the effects of unilateral and bilateral proprioceptive deafferentation of the EOMs on the accuracy of reaching movements directed to visual targets. No significant changes occurred in the mean accuracy (constant error) or variance (variable error) of pointing after unilateral or bilateral deafferentation. We concluded that in normal animals efference copy provides sufficient information about orbital eye position to code space in craniotopic coordinates. PMID- 9744969 TI - Trends in the incidence of myocardial infarction and in mortality due to coronary heart disease, 1987 to 1994. AB - BACKGROUND AND METHODS: To clarify the determinants of contemporary trends in mortality from coronary heart disease (CHD), we conducted surveillance of hospital admissions for myocardial infarction and of in-hospital and out-of hospital deaths due to CHD among 35-to-74-year-old residents of four communities of varying size in the United States (a total of 352,481 persons in 1994). Between 1987 and 1994, we estimate that there were 11,869 hospitalizations for myocardial infarction (on the basis of 8572 hospitalizations sampled) and 3407 fatal coronary events (3023 sampled). RESULTS: The largest average annual decrease in mortality due to CHD occurred among white men (change in mortality, 4.7 percent; 95 percent confidence interval, -2.2 to -7.1 percent), followed by white women (-4.5 percent; 95 percent confidence interval, -0.7 to -8.2 percent), black women (-4.1 percent; 95 percent confidence interval, -10.3 to +2.5 percent), and black men (-2.5 percent; 95 percent confidence interval, -6.9 to +2.2 percent). Overall, in-hospital mortality from CHD fell by 5.1 percent per year, whereas out-of-hospital mortality declined by 3.6 percent per year. There was no evidence of a decline in the incidence of hospitalization for a first myocardial infarction among either men or women; in fact, such hospital admissions increased by 7.4 percent per year (95 percent confidence interval for the change, +0.5 to +14.8 percent) among black women and 2.9 percent per year (95 percent confidence interval, -3.6 to +9.9 percent) among black men. Rates of recurrent myocardial infarction decreased, and survival after myocardial infarction improved. CONCLUSIONS: From 1987 to 1994, we observed a stable or slightly increasing incidence of hospitalization for myocardial infarction. Nevertheless, there were significant annual decreases in mortality from CHD. The decline in mortality in the four communities we studied may be due largely to improvements in the treatment and secondary prevention of myocardial infarction. PMID- 9744971 TI - Antidepressants and the risk of falls among nursing home residents. AB - BACKGROUND: In nursing home residents, the use of tricyclic and other heterocyclic antidepressants is associated with an increased risk of falls. The newer selective serotonin-reuptake-inhibitor antidepressants are largely free of the side effects of the tricyclic agents thought to cause falls and so have been hypothesized to be safer for those at high risk for falls. METHODS: We retrospectively identified an inception cohort of 2428 nursing home residents in Tennessee who were new users of tricyclic antidepressants (665 subjects), selective serotonin-reuptake inhibitors (612 subjects), or trazodone (304 subjects) or nonusers of antidepressants (847 subjects). We ascertained the number of falls during therapy and during a similar follow-up period for nonusers, then calculated the rate ratios for falls with adjustments for an extensive set of potential confounding factors. RESULTS: The new users of each type of antidepressant had higher rates of falls than the nonusers, with adjusted rate ratios of 2.0 (95 percent confidence interval, 1.8 to 2.2) for tricyclic antidepressants, 1.8 (1.6 to 2.0) for selective serotonin-reuptake inhibitors, and 1.2 (1.0 to 1.4) for trazodone. The rate ratios increased with the daily dose for tricyclic antidepressants, reaching 2.4 (95 percent confidence interval, 2.1 to 2.8) for doses of 50 mg or more of amitriptyline or its equivalent, and for the serotonin-reuptake inhibitors, reaching 1.9 (1.7 to 2.2) for 20 mg or more of fluoxetine or its equivalent. The elevated rates of falls persisted through the first 180 days of therapy and beyond. CONCLUSIONS: In this large study of nursing home residents, there was little difference in rates of falls between those treated with tricyclic antidepressants and those treated with selective serotonin reuptake inhibitors. Hence, the preferential use of the newer antidepressants is unlikely to reduce the higher rate of falls among nursing home residents taking antidepressants. PMID- 9744970 TI - High-level chloramphenicol resistance in Neisseria meningitidis. AB - BACKGROUND: Neisseria meningitidis is nearly always susceptible to the penicillins, the cephalosporins, and chloramphenicol. Between 1987 and 1996, however, chloramphenicol-resistant strains were isolated from 11 patients in Vietnam and 1 in France. METHODS: The minimal inhibitory concentration of chloramphenicol was determined for the 12 isolates. The isolates were analyzed by monoclonal-antibody-based serotyping and subtyping, pulsed-field gel electrophoresis, and multilocus enzyme electrophoresis. Bacterial DNA was analyzed by hybridization, the polymerase chain reaction, and sequencing to identify the resistance gene and determine the origin of the resistance. RESULTS: The isolates were resistant to chloramphenicol (minimal inhibitory concentration, > or =64 mg per liter) and produced an active chloramphenicol acetyltransferase. All 12 strains belonged to serogroup B but had a high degree of diversity, and 10 could not be typed with the use of monoclonal antibodies. The nucleotide sequence of the resistance gene and the flanking regions was identical to that of an internal portion of transposon Tn4451 that carries the catP gene in Clostridium perfringens. Moreover, this gene was located in the same genomic site in the chloramphenicol-resistant isolates. CONCLUSIONS: The high-level chloramphenicol resistance that we describe in N. meningitidis isolates is of great concern, since in developing countries, chloramphenicol given intramuscularly is the standard therapy for meningococcal meningitis. The resistance to chloramphenicol is due to the presence of the catP gene on a truncated transposon that has lost mobility because of internal deletions, and the transformation of genetic material between strains of N. meningitidis probably played an important part in the dissemination of the gene. PMID- 9744973 TI - Images in clinical medicine. Human immunodeficiency virus nephropathy and intraglomerular Cryptococcus neoformans. PMID- 9744972 TI - Cushing's syndrome caused by corticotropin secretion by pulmonary tumorlets. PMID- 9744974 TI - Glomerulonephritis. PMID- 9744975 TI - Doxorubicin-induced cardiomyopathy. PMID- 9744977 TI - Death rates from coronary disease--progress and a puzzling paradox. PMID- 9744978 TI - Chloramphenicol resistance in meningococci. PMID- 9744979 TI - Depression in the elderly--falls and pitfalls. PMID- 9744980 TI - The shadowlands--secrets, lies, and assisted reproduction. PMID- 9744982 TI - Effects of menstrual history and use of medications on bone mineral density: the EVOS Study. AB - We have previously shown considerable between-center variation in bone mineral density (BMD) in the 13 EVOS centers that performed bone densitometry on their sex- and age-stratified population samples, after adjusting for weight and age. We have now investigated whether part of the between-center variability may be attributed to between-center variations in the use of medications. Information was collected from 2088 women and 1908 men at baseline on whether the subjects had ever been prescribed calcium, calcitonin, anabolic steroids, fluoride, vitamin D, or glucocorticoids and, for the women, whether they had ever used the oral contraceptive pill (OCP) or hormone replacement therapy (HRT). Each of these variables was fitted into a regression model adjusted for age, height, weight, and center. Only OCP and HRT significantly affected BMD. Those who had ever used OCPs had spinal BMD 0.029 g/cm2 greater than those who had never used them. Users of HRT had higher BMD than nonusers: 0. 037 g/cm2 at the spine, 0.018 g/cm2 at the trochanter, and 0.018 g/cm2 at the femoral neck. As expected, there was a great variation between centers in the use of OCP and HRT, but there were no significant correlations between mean BMD at any site in a given center and the prevalence of OCP or HRT use in that center. The between-center variance in BMD at all three sites remained highly significant after adjusting for treatment (P < 0.001). We conclude that HRT and OCP use are associated with moderate increases in BMD. The geographical variability of BMD in Europe was not explained by treatment with pharmaceuticals. PMID- 9744984 TI - Effects of high-intensity resistance training on bone mineral density in young male powerlifters. AB - The effects of high-intensity resistance training on bone mineral density (BMD) and its relationship to strength were investigated. Lumbar spine (L2-L4), proximal femur, and whole body BMD were measured in 10 male powerlifters and 11 controls using dual-energy X-ray absorptiometry (DXA). There were significant differences in lumbar spine and whole body BMD between powerlifters and controls, but not in proximal femur BMD. A significant correlation was found between lumbar spine BMD and powerlifting performance. These results suggest that high-intensity resistance training is effective in increasing the lumbar spine and whole body BMD. PMID- 9744983 TI - Geographic differences in bone turnover: data from a multinational study in healthy postmenopausal women. AB - Biochemical markers of bone metabolism (bone markers) are used increasingly to monitor response to therapy and may be predictors of bone loss and fractures. The relationship between fracture rates, which differ between countries, and the rate of bone turnover has not been examined. Therefore, we explored the geographic variability of bone turnover in a selected, healthy study population of 619 postmenopausal women, ages 40-61, participating in a clinical trial of raloxifene hydrochloride for osteoporosis prevention. The subjects were distributed among 38 investigative sites in 10 countries (9-211 subjects/country) on four continents (North America, n = 277, Europe, n = 168, Australia, n = 125, and Africa, n = 49). Specimens for serum osteocalcin (OC), bone-specific alkaline phosphatase (BSAP), and urine type I collagen fragment/urinary creatinine ratio (CTX) were handled in a uniform fashion and assayed in a central laboratory. Mean levels of OC (P < 0.001), BSAP (P = 0. 006), and CTX (P < 0.001) varied significantly by country (ANOVA), with the lowest values typically in German and Spanish subjects and the highest in American and Canadian subjects. The consistent pattern and wide ranges of mean bone marker values (OC 1.6-fold, BSAP 1.7-fold, CTX 3.1-fold) between countries suggest clinically significant differences in bone turnover. Geographic differences in bone markers were not explained by the determined potential confounders of age, years posthysterectomy, total serum cholesterol, and serum follicle stimulating hormone (FSH). We conclude that bone marker values vary substantially by country in this selected study population, suggesting systematic geographic differences in bone metabolism that potentially relate to osteoporotic fracture rates. PMID- 9744985 TI - Loss of bone mineral density in women athletes during aging. AB - Total and regional bone mineral density (BMD) by dual-energy-X-ray absorptiometry (DXA) and bone turnover were tested in 50 highly trained women athletes and 21 sedentary control women (18-69 years; BMI < 25 kg/m2). VO2max (ml . kg-1 . min-1) and lean tissue mass (DXA) were significantly higher in the athletes versus controls (both P < 0.0001). Total body BMD did not decline significantly with age in the athletes whereas lumbar spine (L2-L4) BMD approached statistical significance (r = -0.26; P = 0.07). Significant losses of the femoral neck (r = - 0.42), Ward's triangle (r = -0.53), and greater trochanter BMD (r = -0.33; all P < 0.05) occurred with age in the athletes. In the athletes, total body BMD, L2-L4 BMD, and BMD of all sites of the femur were associated with lean tissue mass (r = 0.32 to r = 0.57, all P < 0.05) and VO2max (r = 0.29 to r = 0.48, all P < 0.05). Femoral neck and greater trochanter BMD were higher in the athletes than in controls (both P < 0.05) and lumbar spine and Ward's triangle BMD approached statistical significance (both P = 0.07). Bone turnover was assessed by serum bone-specific alkaline phosphatase (B-ALP), urinary deoxypyridinoline cross-links (Dpd), and urinary aminoterminal cross-linked telopeptides (NTX). There were no relationships between B-ALP or Dpd with age whereas NTX increased with age (r = 0.46, P < 0.05) in the entire group. Levels of B-ALP and NTX were negatively associated with total body, L2-L4, femoral neck, Ward's triangle, and greater trochanter BMD (P < 0.05). B-ALP and Dpd were not significantly different between athletes and controls whereas NTX was lower in the athletes than in controls (P < 0.001). The high levels of physical activity observed in women athletes increase aerobic capacity and improve muscle mass but are not sufficient to prevent the loss of bone with aging. PMID- 9744986 TI - Polymorphism at the Sp 1 binding site in the collagen type I alpha 1 gene does not predict bone mineral density in postmenopausal women in sweden. AB - Polymorphisms at the binding site of the Sp 1 transcription factor of the collagen type I alpha1 gene have recently been suggested to be an important marker for low bone mineral density (BMD) and vertebral fracture in a population of predominantly postmenopausal British women. We examined whether the unfavorable "s" allele was associated with low BMD in 64 patients with primary osteoporosis and in 72 healthy controls. We found no statistically significant differences between COLIA1 genotypes with regard to BMD at the spine and femoral neck. In 36 patients with severe osteoporosis with vertebral fracture the genotype frequencies were similar to that observed in 67 age-matched controls. These data indicate that the Sp 1 polymorphisms in the COLIA1 gene are unlikely to be of clinical value in identifying Swedish subjects who are at risk of postmenopausal osteoporosis. PMID- 9744987 TI - Short-term reproducibility of proximal femur bone mineral density in the elderly. AB - Densitometric measurements are prone to imprecision in elderly subjects and the present study was primarily designed to dissect out the effects of age and bone mineral density on proximal femur dual energy x-ray absorptiometry (DXA) reproducibility. The study comprised 17 elderly women (mean age 74.6 years, range 65-84 years), 13 early postmenopausal women with osteopenia (mean age 56.2 years, range 50-63 years), and 17 elderly men (mean age 73.8 years, range 65-86 years). Each subject was given triplicate proximal femur scans by a QDR 2000 Densitometer (Hologic Inc., Waltham, MA) with repositioning between scans. Because of subject selection in the early postmenopausal women there were no significant differences in bone mineral density (BMD) at any site among the three groups. Despite this, reproducibility errors expressed as either coefficient of variation (CV) % or mean standard deviation (SD) were greater in the elderly subjects, regardless of gender, when compared with the younger female subjects. The variability in measurement errors with age were least marked for the total hip and trochanteric sites. Within the elderly subjects, BMD appeared to exert little influence on measurement errors. We conclude that short-term proximal femur reproducibility is dependent on age-related factors other than BMD. There is no influence of gender on the measurement errors. It is likely that local factors (e.g., hip osteoarthritis) or general frailty may influence repositioning but this needs further exploration. In the meantime, the total hip and trochanteric sites should be used as they provide the most reproducible measurements in the elderly. PMID- 9744988 TI - Effect of region of interest location on ultrasound measurements of the calcaneus. AB - Ultrasound (US) measurements of the calcaneus are usually carried out in a region of interest (ROI) at a fixed site relative to a footplate. Recently, US transmission systems have been developed with imaging capability that enable selection of the position of ROI; the region of measurement is always the area of minimum attenuation in the posterior part of the calcaneus. This study compares measurements of broadband ultrasound attenuation (BUA) and speed of sound (SOS) at the variable ROI of minimum attenuation (ROIv) and at fixed coordinates (ROIf). Ultrasound variables were estimated at ROIv and ROIf in 212 female subjects, including 26 patients with osteoporotic fractures. Among the 186 women without fractures, 63 were classified as having osteoporosis on the basis of their vertebral bone density. Precision of BUA and SOS were better at ROIv than at ROIf. BUA was more highly correlated with bone mineral density (BMD) at the lumbar spine and femoral neck at ROIv than ROIf (r = 0.64 for lumbar spine and 0.68 for femoral neck at ROIv versus 0.50 for lumbar spine and 0.54 for femoral neck at ROIf, P < 0.05 for both comparisons). There were no significant differences between the correlations of SOS with axial BMD at ROIv compared with ROIf. Significant difference was found between the areas under the ROC curve for each ultrasound variable at ROIv and ROIf for both groups of patients, subjects with osteoporosis (area under curve = 0.87 for BUA at ROIv versus 0.82 at ROIf, P < 0.05; area under curve = 0.85 for SOS at ROIv versus 0.81 at ROIf, P < 0. 05), and women with fractures (area under curve = 0.93 for BUA at ROIv versus 0.86 at ROIf, P < 0.05; area under curve = 0.91 for SOS at ROIv versus 0.82 at ROIf, P < 0.05). Ultrasound variables measured at ROIv enable improved reproducibility and significantly better differentiation of diseased subjects from healthy individuals as compared with measurements at ROIf. PMID- 9744989 TI - Polymorphism of insulin-like growth factor I gene and bone mineral density. AB - The polymorphism of insulin-like growth factor-I (IGF-I) gene was examined in Japanese postmenopausal women to analyze the genetic background for osteoporosis. In this study, the dinucleotide (cytosine-adenine; CA) repeat sequence lying upstream of the transcription initiation site of this gene was examined. We named the most frequent allele including (CA) 19 as J allele. There were 6 alleles (J-4 containing 17 CA repeats: (CA)17, [J-2 (CA)18, J (CA)19, J + 2 (CA)20, J + 4 (CA)21, J + 6 (CA)22]) in the Japanese population. The genotype distribution was different from that of Caucasians. There was no different in bone mineral density (BMD) between the group with one or two alleles of each genotype and that without that genotype. When we separate the subjects into three groups having two alleles, one allele, and no alleles, the three subjects who possess the allele 'J 2' in both strands had low BMD (Z score of L2-4; -1.24 +/- 0.56, total body; 0.943 +/- 0.59, mean +/- SE). On the other hand, sequence of IGF-I gene in this study was different from reported sequence of IGF-I gene; that was 2 base pair (bp) deletion following 3'end of CArepeat (-645adenine/-646guanine). The present study showed that there was no association between the microsatellite polymorphism of IGF-I gene and BMD in Japanese postmenopausal women, but some possibility remains that the microsatellite polymorphism of IGF-I gene is useful to detect a kind of particular osteoporosis. PMID- 9744990 TI - Effects of ipriflavone on perialveolar bone formation. AB - The effect of ipriflavone (IP), a synthetic isoflavonoid derivative, on in vivo bone formation was studied in rat perialveolar bone by surgically producing a hole in the mandibular bone. The holes were filled either with powdered IP or with compounds containing no osteoinductive properties such as biostite and Htr (hard tissue replacement). In control animals, the holes were left to heal spontaneously. The animals were killed 3, 28, and 40 days after surgery and a detailed morphological and morphometric study was performed on the perialveolar bone surrounding the wounds. Three days after surgery (inflammatory phase) the bone wounds were occupied by hemorragic and inflammatory cells in both the untreated and IP-treated bone defects. Twenty-eight days after surgery, bone formation was evident with new bone spiculae particularly concentrated in the area of the bone lesion closest to the adjacent periodontal ligament. Morphometric measurements of the areas occupied by new bone showed that the synthesis of perialveolar bone was significantly stimulated by IP. The repair of the bone defects by new bone formation progressed by day 40, but only in the presence of IP were the original holes almost completely repaired. Conversely, biostite and Htr did not influence promotion of new bone formation. In conclusion, the results of the present study are consistent with a role of IP in stimulating osteogenesis and suggest that this compound could represent a potential therapeutic tool to promote repair of injured perialveolar bone. PMID- 9744991 TI - Inhibitory effects of 1,25(OH)2D3 on membrane-associated and cytosolic casein kinase activity in MC3T3-E1 osteoblast-like cells. AB - The secretion of phosphorylated matrix proteins is high in osteoblasts. Phosphorylation of these proteins may be catalyzed by casein kinases (CK), and CK may play an important role in the site of bone mineralization. In this study, we examined the effects of 1, 25(OH)2D3 on CK activities in MC3T3-E1 osteoblast-like cells. Different concentrations (ranging from 10(-7) to 10(-11)M) of 1, 25(OH)2D3 were included in a culture medium. After incubation for various lengths of time, MC3T3-E1 cells were homogenized and segregated into cytosolic (c) and microsomal (m) fractions. To measure CK activity, each fraction was used as an enzyme source to phosphorylate casein. MC3T3-E1 cells showed the highest cCK activity after incubation for 21 days, and showed the highest mCK activity after incubation for 14 days. 1,25(OH)2D3 inhibited mCK activity at the early stage of culture, but inhibited cCK activity at the late stage of culture. In contrast, 1,25(OH)2D3 had a slight stimulatory effect on CK activity in the culture medium of MC3T3-E1 cells. Our data suggest that cCK and mCK may play different roles in the function of osteoblasts, and 1,25(OH)2D3 regulates intracellular and extracellular casein kinase activities related to the function of osteoblasts. PMID- 9744992 TI - The effect of alendronate on cytokine production, adhesion molecule expression, and transendothelial migration of human peripheral blood mononuclear cells. AB - Since both osteoclasts and macrophages belong to the mononuclear phagocytic system it is conceivable that bisphosphonates not only affect bone metabolism but also inflammatory responses. The migration of mononuclear cells into perivascular tissue is a central event in inflammatory reactions. We studied the effects of the aminobisphosphonate alendronate on the transendothelial migration of human peripheral blood mononuclear cells in an in vitro model. Alendronate (at a concentration of 100 microM) significantly increased the percentage of peripheral blood mononuclear cells that migrated through endothelial cell monolayers. Similar results were obtained with another aminobisphosphonate, viz, pamidronate. An overnight treatment of the endothelial cell monolayers with alendronate did not alter the rate of peripheral blood mononuclear cells that subsequently migrated. The overnight cultivation of the peripheral blood mononuclear cells in the presence of alendronate resulted in an increased surface expression of CD54 (intercellular adhesion molecule-1, ICAM-1) in both CD14(+) and CD3(+) cells; in CD14(+) cells also the expression of CD49d (alpha4 subunit of late activation antigen-4, VLA-4) increased after alendronate treatment. Alendronate treatment of peripheral blood mononuclear cells also resulted in an increased production of interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha), and interferon-gamma (IFN-gamma). We conclude that alendronate has a distinct effect on the transendothelial migration of human peripheral blood mononuclear cells in vitro. Alendronate may either directly or indirectly, e.g., by augmenting the production of proinflammatory cytokines, influence the expression of certain adhesion molecules and thereby facilitate transendothelial migration. These effects could be related to the transient leukopenia reported following intravenous administration of relatively high doses of aminobisphosphonates for the treatment of hypercalcemia of malignancy. PMID- 9744993 TI - Osteocyte-bone lining cell system at the origin of steady ionic current in damaged amphibian bone. AB - A wound-generated steady electric current was measured by a two-dimensional vibrating probe system in the metatarsal bones of 22 adult frogs (Xenopus laevis) placed in amphibian Ringer. Inward currents were recorded entering a micrometric hole drilled through the cortex at middiaphyseal level. These steady state currents (mean +/- SD 8.50 +/- 2.77 microA/cm2) last approximately 2 hours, were dependent on the presence of sodium in the incubation medium, were no more detectable after fixation, and were reduced to background level when the cell membranes were solubilized. These results agree with previous recordings of metatarsal bones of weanling mice, under identical conditions. Both results suggest that the measured ionic currents have a cellular origin. Metatarsal bones of adult amphibian were purposely selected for this study because, unlike mammalian bones, their shafts are avascular and only contain an osteocyte-bone lining cell system, as documented by scanning and transmission electron observations. Thus, unlike the data from previous investigations on mammals, the results succeeded in giving the first convincing evidence that the osteocyte-bone lining cell system is the origin of damage-generated ionic currents. As damage exposes bone ionic compartment to plasma, damage-generated ionic currents are representative of ion fluxes at bone plasma interface, and cells at the origin of the current generate the driving force of such fluxes. By demonstrating that osteocytes and bone lining cells are at the origin of the current, this study suggests that the osteocyte-bone lining cell system, though operating as a cellular membrane partition, regulates ionic flow between bone and plasma. Since strain-related adaptive remodeling could also depend on ionic characteristics and flow of the bone fluid through the osteocyte lacuno-canalicular network, the results reported here support the view that osteocyte and bone lining cells may constitute a functional syncytium involved in mineral homeostasis as well as in bone adaptation to mechanical loading. PMID- 9744994 TI - Pasteurella multocida toxin stimulates bone resorption by osteoclasts via interaction with osteoblasts. AB - In this study we used an in vitro assay system with osteoblast and osteoclast co cultures to assess the effect of purified recombinant Pasteurella multocida toxin on bone resorption. Resorption was measured by the release of a telopeptide breakdown product of type I collagen. It was found that P. multocida did not stimulate bone resorption by osteoclasts directly and also did not stimulate bone breakdown via the release of collagenase or other proteases from osteoblasts. During co-culture of osteoblasts and osteoclasts, with cell-cell contact prevented, P. multocida toxin produced no significant effect. Osteoblast conditioned media gave a biphasic effect; low concentrations of P. multocida toxin stimulated bone resorption, whereas 100 ng/ml inhibited resorption by osteoclasts. However, when both cell types were co-cultured with cell-cell contact permitted, P. multocida toxin induced a large concentration-dependent increase in bone resorption over a 7-day period. This suggested that P. multocida toxin causes bone breakdown via an osteoblast-dependent mechanism and that a membrane-bound receptor may be involved. PMID- 9744995 TI - Nicotinamide adenine dinucleotide phosphate oxidase in the formation of superoxide in osteoclasts. AB - Osteoclasts use a variety of chemical agents to degrade bone. One important component of this process is the generation of superoxide. It has been reported that nicotinamide adenine dinucleotide phosphate (NADPH) oxidase is the enzyme responsible for superoxide production in phagocyte; however, the NADPH oxidase present in osteoclasts has not been studied in detail. One of the membrane-bound subunits of the NADPH oxidase is gp91(phox) which represents the rate-limiting component for the formation of the NADPH oxidase complex. This study was designed to demonstrate the presence of gp91(phox) in individual osteoclasts using the RT PCR technique developed for limited numbers of cells. Compared with white cells, 1.8 times the amount of gp91(phox) mRNA was found in osteoclasts. This difference may be related to the size of the osteoclast and the multiple nuclei present. The presence of gp91(phox) in osteoclasts was confirmed at protein level by immunocytochemistry. Osteoclastic superoxide generation is inhibited by diphenylene iodonium, a specific inhibitor of the NADPH oxidase. These studies suggest that superoxide generation by osteoclasts correlates with the activity of NADPH oxidase. PMID- 9744996 TI - Expression of the mRNA for types I and II interleukin-1 receptors in dental tissues of mice during tooth development. AB - Interleukin-1 (IL-1) can exert its pleiotropic effects on nearly every tissue by binding to its cognate receptor. Two types of IL-1 receptors have been identified. A large number of cell types have been shown to possess IL-1 receptors in vitro and in vivo, but few studies have addressed the question of expression in dental tissues in vivo. Using in situ hybridization in normal newborn, young and adult mice, we have examined the cellular distribution of both types of IL-1 receptors in dental tissues. In the ameloblast layer of incisors and molars, the mRNA for the type I IL-1 receptor (IL-1RI) and the type II IL-1 receptor (IL-1RII) was detected at the presecretory stage. The expression level markedly increased and remained during amelogenesis at the secretory stage. At the maturation stage, however, the transcripts for both IL-1RI and -II mRNA disappeared. Expression of IL-1RI and -II mRNA was also observed in odontoblasts after crown morphogenesis had been completed, and continued in these cells during dentinogenesis. No transcripts were detected in stratum intermedium cells and other cells in dental follicle, stellate reticulum, dental papilla, or pulp. Additionally, both types of IL-1R mRNA were also detected in osteoclasts on surfaces of alveolar bone. These results demonstrated for the first time that enamel-secreting ameloblasts and dentine-secreting odontoblasts express IL-1RI and -II mRNA, suggesting that IL-1 plays a regulatory role in the function of ameloblasts and odontoblasts during tooth development of mice. PMID- 9744997 TI - Induction of bone formation using a recombinant adenoviral vector carrying the human BMP-2 gene in a rabbit spinal fusion model. AB - Bone marrow-derived mesenchymal stem cells are pluripotential cells that have the capacity to differentiate into an osteoprogenitor line. It has been demonstrated that BMP-2 can enhance this differentiation process. In an attempt to prolong the transforming effect of BMP-2, we used an adenoviral vector carrying the human BMP 2 gene to transduce marrow-derived mesenchymal stem cells of New Zealand white rabbits. Assays on tissue culture demonstrated that these cells indeed produced the BMP-2 protein. These transduced stem cells were then autologously reimplanted into the donor rabbits. The cells were placed in the intertransverse process area of five rabbits. In one out of the five rabbits, this resulted in the production of new bone which was demonstrable on both radiographic and histologic examination. We conclude that it is possible to successfully transduce mesenchymal stem cells with the gene for BMP-2 such that these cells will produce the BMP-2 protein in vitro. Further, the transduction results in transformation of these cells into an osteoprogenitor line capable of producing bone in vivo. These data suggest the feasibility of employing gene therapy using recombinant adenoviral vectors as a tool for enhancing spine fusion. Further work to improve the fidelity and longevity of the gene transfer is warranted. PMID- 9744998 TI - Modulation of the plasma membrane Ca2+ pump. AB - The plasma membrane calcium pump, which ejects Ca2+ from the cell, is regulated by calmodulin. In the absence of calmodulin, the pump is relatively inactive; binding of calmodulin to a specific domain stimulates its activity. Phosphorylation of the pump with protein kinase C or A may modify this regulation. Most of the regulatory functions of the enzyme are concentrated in a region at the carboxyl terminus. This region varies substantially between different isoforms of the pump, causing substantial differences in regulatory properties. The pump shares some motifs of the carboxyl terminus with otherwise unrelated proteins: The calmodulin-binding domain is a modified IQ motif (a motif which is present in myosins) and the last 3 residues of isoform 4b are a PDZ target domain. The pump is ubiquitous, with isoforms 1 and 4 of the pump being more widely distributed than 2 and 3. In some kinds of cells isoform 1 or 4 is missing, and is replaced by another isoform. PMID- 9744999 TI - Band 3-mediated flip-flop and phosphatase-catalyzed cleavage of a long-chain alkyl phosphate anion in the human erythrocyte membrane. AB - In pursuit of the characterization of the recently discovered flippase mode of operation of the anion transporter (band 3, AE1) of the human erythrocyte membrane, the transbilayer translocation (flip) of a fluorescently labeled, membrane-intercalated long-chain alkyl phosphate, 10-(alpha-napthyl)-1-decyl phosphate (NDP) was investigated. In contrast to the alkyl sulfonates and esters of phosphatidic acid studied as yet, NDP moves exclusively via band 3. NDP is, however, dephosphorylated at the inner membrane surface by a cytoplasmic phosphatase likely to interact specifically with endofacial membrane structures of the erythrocyte. This phosphatase shares characteristic inhibitor sensitivities with protein tyrosine phosphatases present in the erythrocyte interior. Vanadate as an inhibitor of NDP dephosphorylation provided a means to study the kinetic properties and patterns of inhibition (by inhibitors of anion exchange) and stimulation (by proteolysis of band 3 and aliphatic alcohols) of the flip of NDP. NDP is also an inhibitor of the exchange of hydrophilic anions via band 3, while hydrophilic anions interfere with the flip of NDP. The results are compared with the characteristics of the flip, via Band 3, of other amphiphilic anions and of the exchange of hydrophilic anions. Attempts are presented to understand the low flip rate of long-chain amphiphilic anions on the basis of their molecular properties and the thermodynamics of the "transition state" of the flip process. PMID- 9745000 TI - Urea inhibits the Na-K pump in human erythrocytes. AB - Recent studies have established that urea alters the activity of several volume sensitive cation transport pathways. However, it has remained unclear whether urea has any effect on transport pathways that are not volume-sensitive. We examined the effect of urea on Na-K pump in the human erythrocytes. In cells from nine subjects, 500 mM urea inhibited 52 +/- 10% of the pump activity measured as the ouabain-sensitive (OS) K influx. Urea inhibited the OS K influx reversibly, in a concentration-dependent manner. [3H] oubain binding, a measure of the number of Na-K pump sites remained unchanged with urea. Urea decreased the Vmax for ouabain-sensitive K influx, but did not alter the apparent K(m) for external K. Furthermore, urea did not alter the apparent K(m) for intracellular Na. The ion turnover per pump site was decreased in the presence of urea. Thus, physiologically relevant urea concentration inhibit the Na-K pump in human erythrocyte. The inhibition of the Na-K pump by urea suggests that the effects of urea may not be limited to volume-sensitive transporters, but may be more widespread. PMID- 9745002 TI - Voltage-dependent conductance changes in a nonvoltage-activated sodium current from a mast cell line. AB - Nonexcitable cells do not express voltage-activated Na+ channels. Instead, selective Na+ influx is accomplished through GTP-activated Na+ channels, the best characterized of which are found in renal epithelia. We have described recently a GTP-dependent Na+ current in rat basophilic leukemia (RBL) cells that differs from previous reported Na+ channels in several ways including selectivity, pharmacology and mechanism of activation. In this report, we have investigated the biophysical properties of the RBL cell Na+ current using the whole cell patch clamp technique. Following activation by 250-500 microM GTP gamma S, hyperpolarizing steps to a fixed potential (-100 mV) from a holding potential of 0 mV evoked transient inward Na+ currents that declined during the pulse. If the holding potential was made more positive (range 0 to +100 mV), then the amplitude of the transient inward current evoked by the hyperpolarization increased steeply, demonstrating that the conductance of the channels was voltage dependent. Using a paired pulse protocol (500 msec pulses to -100 mV from a holding potential of 0 mV), it was found that the peak amplitude of the current during the second pulse became larger as the interpulse potential became more positive. In addition, increasing the time at which the cells were held at positive potentials also resulted in larger currents, indicating a time-dependent conductance change. With symmetrical Na+ solutions, outward currents were recorded at positive potentials and these demonstrated both a time- and voltage dependent increase in conductance. The results show that a nonvoltage activated Na+ channel in an electrically nonexcitable cell undergoes prominent voltage dependent transitions. Possible mechanisms underlying this voltage dependency are discussed. PMID- 9745001 TI - The action of blocking agents applied to the inner face of Ca(2+)-activated K+ channels from human erythrocytes. AB - The actions of clotrimazole and cetiedil, two drugs known to inhibit the Gardos channel, have been studied on single intermediate conductance calcium-activated potassium (IKCa) channels in inside out patches from human red blood cells, and compared with those of TEA and Ba2+ applied to the cytoplasmic face of the membrane. TEA produced a fast block which was observed as a reduction in the amplitude of the single channel current. This effect was weakly voltage dependent with the fraction of the membrane potential sensed by TEA at its binding site (delta) of 0.18 and a Kd at 0 mV of 20.5 mM. Ba2+ was a very potent blocker of the channel, breaking the single channel activity up into bursts, inter-spersed with silent periods lasting several seconds. The effect of Ba2+ was very voltage sensitive, delta = 0.44, and a Kd at 0 mV of 0.15 microM. Clotrimazole applied to the inner face of the membrane at a concentration < or = 1 microM produced a slow block resulting in bursts of channel activity separated by quiescent periods lasting many seconds. The effect of clotrimazole was mimicked by a quaternary derivative UCL 1559, in keeping with an action at the cytoplasmic face of the channel. A high concentration of cetiedil (100 microM) produced only a weak block of the channel. The kinetics of this action were very slow, with burst and inter burst intervals lasting several minutes. While inhibition of the Gardos channel by cetiedil is unlikely to involve an intracellular site of action, if clotrimazole is able to penetrate the membrane, part of its effect may result from binding to an intracellular site on the channel. PMID- 9745004 TI - Effects of mechano-gated cation channel blockers on Xenopus oocyte growth and development. AB - The putative role(s) of a mechanically gated (MG) cation channel in Xenopus oocyte growth, maturation, fertilization and embryogenesis has been examined. Using a pharmacological approach, we have tested the effects of the MG channel blockers, gadolinium, gentamicin and amiloride on the above developmental events. Our results indicate that oocyte maturation, fertilization and early embryogenesis (up to the free-swimming stage 45) can proceed normally in the presence of concentrations of agents that either completely abolish (i.e., > or = 10 microM Gd3+) or partially block (i.e., 1 mM gentamicin) single MG channel activity as measured by patch-clamp recording. However, we also find that higher concentrations of Gd3+ (> or = 50 microM) can lead to an increased percentage (> 20%) of axis-perturbed embryos compared with control (< 1%) and that amiloride (0.5 mM) reduces the success of fertilization (from 100% to < 50%) and increases mortality (by approximately 75%) in developing embryos. Furthermore, we find that all three agents inhibit oocyte growth in vitro. However, their order of effectiveness (amiloride > gentamicin > Gd3+) is opposite to their order for blocking MG channels (Gd3+ >> gentamicin > amiloride). These discrepancies indicated that the drugs effects occur by mechanisms other than, or in addition to, MG channel block. Our results provide no compelling evidence for the idea that MG channel activity is critical for development in Xenopus. This could mean that there are other mechanisms in the oocyte that can compensate when MG channel activity is blocked or that the protein that forms the channel can undergo additional interactions that result in a function insensitive to MG channel blockers. PMID- 9745003 TI - 1-anilino-8-naphtalene sulfonate probes a gastric HK-ATPase potassium site whose access requires ionophores. AB - 1-anilino-8-naphtalenesulfonate (ANS) is a hydrophobic dipole previously used to demonstrate that the proton for potassium exchange by the gastric HK-ATPase is electroneutral. In this paper, we demonstrate that ANS binds to gastric membranes and probes conformational changes of the HK-ATPase independently of any active H for K exchange. Conformational changes require the presence of potassium valinomycin and are not triggered by sodium. Potassium effect is enhanced by ATP, in the presence and in the absence of magnesium and, by ADP, in the presence of magnesium. Labeling of the pig HK-ATPase K518 by fluorescein-5-isothiocyanate inhibits the enzyme activity and knocks out the ATP effect on ANS fluorescence. Scherring 28080 and the monoclonal antibody 95-111, two competitive inhibitors of K-activated ATPase dephosphorylation, do not modify K-effect on ANS fluorescence but inhibit ATP effects. This supports that ANS does not probe K-site between the H1-H2 loop. Treatment of gastric membranes with trypsin does not inhibit the ANS response to potassium but does inhibit the response to ATP. This suggests that the ATP site inducing the ANS response is cytoplasmic and the potassium site is intramembranous. Titration reveals that one mole of ANS interacts with one mole of ATPase. We suggest that ANS probes a hydrophobic potassium site of gastric ATPase and that addition of ATP and ADP-Mg embed that site. PMID- 9745005 TI - The functional surface charge density of a fast K channel in the myelinated axon of Xenopus laevis. AB - The action of Mg2+ on the putative xKv1.1 channel in the myelinated axon of Xenopus laevis was analyzed in voltage clamp experiments. The main effect was a shift in positive direction of the open probability curve (16 mV at 20 mM Mg2+), calculated from measurements of the instantaneous current at Na reversal potential after 50-100 msec steps to different potentials. The shift was measured at an open probability level of 25% to separate it from shifts of other K channel populations in the nodal region. The results could be explained in terms of screening effects on fixed charges located on the surface of the channel protein. Using the Grahame equation the functional charge density was estimated to -0.45 e nm-2. Analyzing this value, together with previously estimated values from other K channels, with reference to the charge of different extracellular loops of the channel protein, we conclude that the loop between the transmembrane S5 segment and the pore forming P segment determines the functional charge density of voltage-gated K channels. PMID- 9745006 TI - Divalent cation effects on the Shaker K channel suggest a pentapeptide sequence as determinant of functional surface charge density. AB - The effects of the divalent cations strontium and magnesium on Shaker K channels expressed in Xenopus oocytes were investigated with a two-electrode voltage-clamp technique. 20 mM of the divalent cation shifted activation (conductance vs. potential), steady-state inactivation and inactivation time constant vs. potential curves 10-11 mV along the potential axis. The results were interpreted in terms of the surface charge theory, and the surface charge density was estimated to be -0.27 e nm-2. A comparison of primary structure data and experimental data from the present and previous studies suggests that the first five residues on the extracellular loop between transmembrane segment 5 and the pore region constitutes the functional surface charges. The results further suggest that the surface charge density plays an important role in controlling the activation voltage range. PMID- 9745007 TI - A pro-B-cell stage characterized by germline Ig transcription without surrogate light chain expression. AB - B-cell commitment is characterized by the expression of specific membrane proteins and the rearrangement and expression of immunoglobulin (Ig) heavy (H) and light (L) chain genes. At early stages of B-cell development, unrearranged Ig loci are transcribed, which correlates with these regions becoming accessible for Ig gene rearrangement. Some germline transcripts can be translated into protein and potentially play a role in cell signaling during B-cell development. In this report an early stage in human B-cell development is characterized using Epstein Barr virus (EBV)-transformed cell lines from patients with a severe combined immunodeficiency (SCID). These lines were shown to produce germline constant (C) gene transcripts from the IGH and IGK loci. We demonstrate here that these cells are committed to the B-cell lineage as substantiated by expression of CD79a and CD79b. No surrogate light chain (SLC) gene transcription was detected, indicative of a very early differentiation stage. From these cell lines two types of germline IgV gene transcripts were isolated: a transcript containing the IGKV4-1 gene and a germline IGHV-1 transcript nearly identical to IGHV1/OR15-1 (HC15-1, DP-1), an orphon VH gene on chromosome 15. Germline VH transcripts originating from the VH locus on chromosome 14 could not be detected. It is of interest that, apart from Ig V and C genes (non-functional), V genes that reside outside the Ig locus are a target for the transcription factors that are postulated to initiate Ig gene rearrangement early in B-cell development. PMID- 9745008 TI - Patterns of reticulate evolution for the classical class I and II HLA loci. AB - Some alleles of the major histocompatibility complex (MHC) genes have a reticulate pattern of evolution, probably resulting from the exchange of segments by gene conversion or recombination. Here we compare the extent and patterns of reticulate evolution among the classical class I and class II loci of the human MHC using the recently developed compatibility and partition matrix methods. A complex pattern is revealed with substantial differences among loci in the extent and pattern of reticulation. Extremely high levels of reticulation are observed at HLA-B and HLA-DPB1, high levels at HLA-A and HLA-DRB1, moderate levels at HLA C and HLA-DQB1, and low levels at HLA-DQA1. The reticulate events are concentrated in the exons encoding the highly variable, peptide-binding domains, suggesting that the sequence combinations produced by these events are maintained by natural selection. PMID- 9745009 TI - A geometric study of the amino acid sequence of class I HLA molecules. AB - HLA class I alleles are studied by representing them in a metric space where each dimension corresponds to each one of the amino acid positions. Their similarity in reference to their ability to present peptides to T cells is then evaluated by calculating the correlation matrix between the amino-acid-composition tables (or binding affinity tables) for the sets of peptides presented by each allele. This correlation matrix is considered an empirical similarity matrix between HLA alleles, and is modeled in terms of possible structures defined in the metric space of HLA class I amino acid sequences. These geometric structures are adequate models of the peptide-binding data currently available. The following clusters of HLA class I molecules are identified in reference to their ability to present peptides: Cluster I) HLA-A3/ HLA-A11/ HLA-A31/ HLA-A33/ HLA-A68; Cluster II) HLA-B35/ HLA-B51/ HLA-B53/ HLA-B54/ HLA-B7; and Cluster III) HLA-A29/ HLA B61/HLA-B44; the last cluster showing possible similarities between alleles from different loci. In modeling these natural clusters, the geometric structures with more predictive power confirm the importance of those positions in the peptide binding groove, particularly those in the B pocket. In addition, other positions (46, 79, 113, 131, 144, and 177) appeared to bear some relevance in determining which peptides can be presented by which HLA alleles. PMID- 9745010 TI - A genetic linkage map of rat chromosome 20 derived from five F2 crosses. PMID- 9745011 TI - Beta 2-microglobulin of ictalurid catfishes. PMID- 9745012 TI - Limited polymorphism in the first domain of the rat MHC class II RT1-D molecule. PMID- 9745013 TI - HLA-A26 subtype A pockets accommodate acidic N-termini of ligands. PMID- 9745014 TI - Cloning and sequencing of cDNA encoding mouse cytohesin-1. PMID- 9745015 TI - Sequence analysis of DA and Sprague Dawley rat T-cell receptor beta-chain promoters. PMID- 9745016 TI - Cloning and characterization of sheep (Ovis aries) immunoglobulin alpha chain. PMID- 9745017 TI - Cell-division-cycle defects associated with fission yeast pre-mRNA splicing mutants. AB - We have isolated six new pre-mRNA splicing mutants (prp) from a collection of temperature-sensitive (ts-) Schizosaccharomyces pombe strains. The prp mutants are defective in the splicing of both messenger RNA and U6 small nuclear RNA precursors. A single recessive mutation is responsible for both the ts- growth and the splicing phenotypes in each of the prp mutants. The six prp mutations are unlinked and fall into separate complementation groups. Two are allelic with the previously described prp3 and prp4 mutations; the remaining four define the new alleles prp5-1, prp6-1, prp7-1, and prp9-1. The six mutants exhibit three splicing phenotypes: accumulation of unspliced precursor at the restrictive but not at the permissive temperature; accumulation of unspliced precursor at both the permissive and restrictive temperatures; and accumulation of unspliced precursor, the intron-exon lariat intermediate, and the intron lariat final product. In addition to their aberrant splicing phenotypes, the prp5-1 and prp6-1 mutants express classical cell-division-cycle defects, while prp7-1 exhibits an unusual hyphal morphology. These results suggest a connection between pre-mRNA splicing and the control of cell division in fission yeast. PMID- 9745018 TI - The essential schizosaccharomyces pombe cdc23 DNA replication gene shares structural and functional homology with the Saccharomyces cerevisiae DNA43 (MCM10) gene. AB - The fission yeast cdc23 gene is required for correct DNA replication: cdc23 mutants show reduced rates of DNA synthesis and become elongated after cell-cycle arrest. We have cloned the Schizosaccharomyces pombe cdc23 gene by complementation of the temperature-sensitive phenotype of cdc23-M36 and confirmed the identity of the gene by integrative mapping. Analysis of the DNA sequence reveals that cdc23 can encode a protein of 593 amino acids (Mr=67 kDa) with 22% overall identity and many structural homologies with the product of the Saccharomyces cerevisiae DNA43 (MCM10) gene which is required for correct initiation of DNA synthesis at chromosomal origins of replication. Construction of a cdc23 null allele has established that the cdc23 gene is essential for viability, with cdc23 deletion mutant spores germinating but undergoing arrest with undivided nuclei in the first or second cell cycle. The S. pombe cdc23 gene on an expression plasmid is able to complement the S. cerevisiae dna43-1 mutant. These structural and functional homologies between two distantly related species suggest that cdc23 and DNA43 may represent genes for a conserved essential eukaryotic DNA replication function. PMID- 9745019 TI - ras1 and pat1 alleles interact to quantitatively and qualitatively alter conjugation in fission yeast. AB - To identify novel components of ras1+ signalling in Schizosaccharomyces pombe, extragenic suppressors of the mating defect of ras1 effector mutants were isolated. A novel allele of pat1, pat1-e1, was isolated that increases the mating of ras1-D43E mutants to near wild-type levels but does not suppress the mating defect of ras1-I41M, ras1-Y37F, or ras1-Y45I mutants. This allele-specific suppression is not a characteristic of all pat1 alleles since pat1-3 and pat1-114 partially and equally suppress ras1-D43E and ras1-I41M mutants. Analysis of mating cultures showed that ras1-D43E and pat1-e1 interact to qualitatively alter the mating response. While pat1-e1 ras1-D43E cells were delayed in agglutination, cell-cycle delay, and mat1-Pm transcription, they induce mat1-Mc at the same time and mate more rapidly than other mating cultures. These results suggest that pheromone signalling, but not nutritional signalling, is delayed in pat1-e1 ras1 D43E cells. We hypothesize that this delay causes an elevated pheromone response and thus suppression of the mating defect of the ras1-D43E mutant by pat1-e1. PMID- 9745020 TI - Ace2p, a regulator of CTS1 (chitinase) expression, affects pseudohyphal production in Saccharomyces cerevisiae. AB - Some diploid strains of Saccharomyces cerevisiae can grow both as a spherical yeast form and as a filamentous pseudohyphal form. Most yeasts capable of forming pseudohyphae possess a functional FLO8 gene. We show that disrupting the ACE2 transcription factor results in the production of pseudohyphae in a flo8-1 background. Disrupting the CTS1 (chitinase) gene also produces pseudohyphal growth in this background, but at a reduced level. Invasion of solid media by haploid and diploid cells is increased in ACE2 disruptions, but the diploids adhere poorly to the agar. Sigma1278b-derived strains, which generally produce pseudohyphae, have about 30-fold lower chitinase activity than other strains. PMID- 9745021 TI - Identification of polymorphic mutant alleles of CaMDR1, a major facilitator of Candida albicans which confers multidrug resistance, and its in vitro transcriptional activation. AB - CaMDR1 (Candida albicans Multi Drug Resistance) encodes a major facilitator whose expression in Saccharomyces cerevisiae confers resistance to several unrelated drugs. We describe here the identification and molecular characterization of seven mutant alleles of CaMDR1 (CaMDR1-1 to 1-7). The complete sequencing of CaMDR1 alleles revealed several in-frame point mutations leading to a change in amino-acid residues where insertion/replacement of an aspartate residue in a serine-asparagine-aspartate-rich domain was most noteworthy. Interestingly, these alleles showed a distinct drug resistance profile. The expression of CaMDR1, or of its alleles, in C. albicans cells was enhanced by benomyl, methotrexate and several other unrelated drugs, and was more pronounced in at least one of the azole-resistant clinical isolates. PMID- 9745022 TI - Evidence for circular transposition derivatives from the fungal hAT-transposon Restless. AB - The structure and function of eukaryotic hAT-transposons has already been elucidated; however, their transposition mechanism is barely understood. We recently have discovered Restless, a fungal member of the hAT-transposon family, which shows transposition in its host Tolypocladium inflatum. Investigations of two strains from T. inflatum, carrying either about 15 copies or only a single copy of the Restless element, indicate the presence of circular transposition intermediates. Using PCR technology, amplicons were identified which carry the joined end of the Restless transposon fused at its inverted repeats. All of eight sequenced PCR fragments contained the intact transposon ends with a short insertion of 1-93 bp of genomic DNA. Remarkably, one of the discovered genomic sequences matches a previously described integration site. Our data are discussed with respect to the transposition mechanism and the horizontal transfer of hAT transposons. PMID- 9745023 TI - Characterization of novel extrachromosomal DNA from giant-celled marine green algae. AB - Cloned HinfI fragments of the plasmid-like 2.2-kb DNA from the green alga Ernodesmis verticillata (Kutzing) Borgesen hybridized solely to single or double bands within the 2.2-kb DNA in genomic Southern blots. Heterologous probes for nuclear and chloroplast genes hybridized only to high-molecular-weight (HMW) DNA. Thus, the low-molecular-weight (LMW) DNA is extrachromosomal and lacks extensive homology to nuclear or chloroplast genes. There was cross-hybridization to LMW DNA from other Caribbean isolates of E. verticillata, but not to that from a Pacific isolate. Under reduced stringency, cross-hybridization to LMW DNA from the related green alga Boergesenia forbesii (Harvey) Feldmann was also observed, suggesting that the LMW DNA may have a common origin and/or function in these algae. Six out of sixteen unique clones hybridized to discrete bands in northern blots, indicating that the LMW DNA may be actively transcribed in vivo. Four of the putatively transcribed clones have regions with significant deduced amino acid sequence identity to psa and psb gene products, implying that the plasmid like molecules might have originated from chloroplast DNA. Sequencing data also indicated a high G/C content, as well as the presence of frequent tandem and direct repeats in many of the cloned fragments. Sequencing and restriction analyses suggest that most of the cloned fragments are portions of different DNA molecules, providing evidence that the 2.2-kb extrachromosomal DNA in Ernodesmis is novel in that it represents a fairly heterogeneous population of molecules. PMID- 9745024 TI - The rbcL gene from the non-photosynthetic parasite Lathraea clandestina is not transcribed by a plastid-encoded RNA polymerase. AB - In the plastome of the obligate root-parasitic plant, Lathraea clandestina, the rbcL gene has been maintained and is expressed, despite the reduced size and gene content of the plastid genome. Some of the plastid genes involved in translation (e. g. transfer RNAs, ribosomal RNAs and ribosomal proteins) have been sequenced and still appear to code for functional ribosomal components. Indeed, the 16S rRNA and rpl20 genes are expressed whilst other necessary tRNA and ribosomal protein-encoding genes have probably been deleted or truncated. Although obtained by PCR, the four rpo genes for Escherichia coli-like plastid encoded RNA polymerase appear to be pseudogenes. Nevertheless, the rbcL gene, with a "-10, 35" prokaryotic-like promoter, is still transcribed. In contrast to photosynthetic plants, rbcL transcripts in Lathraea are larger in their 5' region and cover the prokaryotic-like promoter. The transcription initiation site is located near the ATG start codon of the atpB pseudogene. Similarity to non consensus E. coli-like plastid promoters suggests that rbcL transcription is driven by a nuclear-encoded RNA polymerase. PMID- 9745026 TI - Editing and translation of ribosomal protein S13 transcripts: unedited translation products are not detectable in maize mitochondria. AB - Maize mitochondrial transcripts for the ribosomal protein S13 gene (rps13) have six C- to -U editing sites, and each nucleotide conversion causes a change in the amino acid specified by the effected codons. Sequence analysis of 30 cDNA clones indicated that 73% of the cDNAS were edited at all six sites and 3% were completely unedited. Antibodies were produced against synthetic peptides that corresponded to unedited or edited translation products at editing sites V and VI (80% and 83% edited, respectively). Antibody preparations were purified that selectively recognized the edited or unedited forms of the epitope. The antibody preparations were highly sensitive to the amino-acid residue encoded at editing site VI, but relatively insensitive to the residue encoded at editing site V. Immunological analyses demonstrated that the edited translation product accumulated as a ribosomal protein, but that the unedited translation product was not detected in the mitochondrion, in the ribosomal fraction, or in a post ribosomal supernatant. These results, taken together with other studies which demonstrated that incompletely edited transcripts are incorporated into polyribosomes, suggest that incompletely edited transcripts may be translated, but polypeptides encoded by incompletely edited RNAs may be unstable and, consequently, fail to accumulate. PMID- 9745025 TI - Transcription of plastid-derived tRNA genes in rice mitochondria. AB - Previous investigations located nine of the genes for rice tRNAs on plastid (pt) derived sequences in mitochondrial DNA. In the present study, we examined whether these genes were also transcribed in rice mitochondria. Northern-blot hybridization revealed that seven of these genes (trnC, trnF, trnH, trnM, trnN, trnS and trnW) are transcribed and are precisely processed to mature tRNAs. One of the other two genes (trnP) is transcribed but cannot be processed efficiently, while the other (trnR), which has 100% identity to the original plastid tRNAArg gene, is not transcribed in rice mitochondria. These results suggest that seven of the nine pt-derived tRNAs may be utilized for the biosynthesis of protein in plant mitochondria. PMID- 9745027 TI - Genetic mapping of telomeric DNA sequences in the maize pathogen Cochliobolus heterostrophus. AB - Numerous polymorphic bands were observed when genomic DNA of Cochliobolus heterostrophus was cut with restriction enzymes and hybridized with a telomere specific probe. Bal31 digestion demonstrated that the majority of the bands were from the chromosome termini and thus constitute telomeres. However, numerous hybridizing sequences in the strain B30.A3.R.45 were Bal31 exonuclease insensitive and thus located at least 600 bases internal to the chromosome ends. Segregation analysis of the polymorphic bands identified nine telomeres; seven were linked to other markers and thus permitted the proper orientation of linkage groups with respect to their chromosome ends. Three internal, telomere-homologous sequences, at least one of which may be subtelomeric, were also mapped. PMID- 9745029 TI - Electronic data submissions to MGD from 12/15/97 to 5/26/98 PMID- 9745028 TI - Gibberellin biosynthesis in Gibberella fujikuroi: cloning and characterization of the copalyl diphosphate synthase gene. AB - The gene coding for copalyl diphosphate synthase (CPS), which represents the first gene of the gibberellin pathway, was isolated from the rice pathogen Gibberella fujikuroi. This fungus is used commercially for the production of gibberellic acid and related gibberellins. CPS is a terpene cyclase which catalyzes the first specific step of the gibberellin (GA) pathway as it branches off from the general isoprenoid (biosynthetic) pathway at geranylgeranyl disphosphate (GGDP). A cDNA fragment of the cps gene from the fungus G. fujikuroi was amplified by RT-PCR using oligonucleotides based on amino-acid sequences which were conserved between the plant CPSs and the bifunctional CPS/KS of the fungus Phaeosphaeria sp. L487. A 588-bp fragment obtained with nested PCR was used to isolate the corresponding genomic clone of the cps gene from the wild type lambda-library. This gene consists of three exons and two introns. The three exons are 2877 bp long and encode 959 amino-acid residues. The protein shares 48% identity with the bifunctional Phaeosphaeria sp. L487 FCPS and between 16% and 18% identity to the corresponding plant CPSs. Expression of the G. fujikuroi cps gene is strongly enhanced under conditions optimized for gibberellin biosynthesis and is reduced when high amounts of ammonium are present in the medium. Gene disruption, followed by gibberellin assays and Southern-blot analysis of the transformants, demonstrated clearly that the cloned gene has the expected function in the biosynthesis of fungal gibberellins. PMID- 9745030 TI - Dominant optic atrophy: exclusion and fine genetic mapping of the candidate gene, HRY. AB - Autosomal dominant optic atrophy (OPA1) maps to Chromosome (Chr) 3q28, and the disease interval has been refined to within 1.4 cM, flanked by the markers D3S3669 and D3S3562. HRY, the human homolog of the Drosophila segmentation gene, hairy, maps by in situ hybridization to the chromosomal region 3q28-q29. We screened for mutations in HRY in 36 patients from 18 pedigrees with dominant optic atrophy and a group of normal control individuals. Heteroduplex mutation analysis and direct sequencing of all four coding exons and one upstream putative untranslated exon were performed. No disease-associated sequence alterations were identified. A polymorphism in the untranslated region of exon 2 was found, with four alleles. PCR amplification of this part of exon 2 in four of the pedigrees affected by autosomal dominant optic atrophy mapping to chromosome 3q, followed by haplotype analysis, showed recombination between HRY and OPA1 in one pedigree. This allows us to genetically position HRY in relation to known microsatellite markers in the region, placing HRY telomeric to marker D3S3562 and centromeric to D3S1305. This is outside the published critical disease interval for dominant optic atrophy. We have, therefore, excluded HRY as the gene for dominant optic atrophy by sequence analysis, mapped it genetically, and identified a polymorphism in our population. PMID- 9745031 TI - Structure and function correlations at the imprinted mouse Snrpn locus. AB - The human SNRPN gene maps within Chromosome (Chr) 15q11-q13, the region responsible for Prader-Willi syndrome (PWS) and Angelman syndrome (AS). As one of several 15q11-q13 transcripts expressed from the paternal allele-only, SNRPN is a candidate gene to explain at least some of the PWS phenotype in human and in genetic mouse models. The promoter and first exon of the SNRPN gene also correspond to an imprinting center element responsible for resetting of the maternal to paternal imprints within 15q11-q13 during spermatogenesis. Through characterization of the imprinted murine Snrpn locus in mouse Chr 7C, we have found that the gene structure is very similar to the human, with ten conserved exons spanning 22 kb, the last seven of which are tightly clustered. The promoter of Snrpn is differentially methylated in ES cells and adult tissues, supporting a role for DNA methylation at this site in somatic establishment and/or maintenance of Snrpn imprinting. The first intron of the mouse and human genes contains structurally conserved G-rich clustered repeats which may play a role in establishing DNA methylation patterns associated with imprinting of this gene. On the basis of the conserved structural and imprinted features of the human SNRPN and mouse Snrpn genes, we suggest that imprinting mechanisms are conserved between human and mouse. PMID- 9745032 TI - A major quantitative trait locus co-localizing with cholecystokinin type A receptor gene influences poor pancreatic proliferation in a spontaneously diabetogenic rat. AB - The Otsuka Long-Evans Tokushima Fatty (OLETF) rat is an animal model for obese type, non-insulin-dependent diabetes mellitus (NIDDM) in humans. The OLETF rat has poor capacity for pancreatic proliferation, which may be the critical pathogenetic event in NIDDM development. Our investigation was designed to identify quantitative trait loci (QTLs) responsible for poor pancreatic proliferation by examining compensatory proliferation of the pancreatic remnant after partial pancreatectomy and performing a genome-wide scan in an F2 intercross obtained by mating the OLETF and the Fischer-344 (F344) rats. We identified a highly significant QTL on rat Chromosome 14 with a maximum lod score of 16.7, which accounts for 55% of the total variance. The QTL co-localizes with the gene encoding cholecystokinin type A receptor (CCKAR) which is likely to mediate the trophic effect of cholecystokinin on pancreas and is defective in the OLETF rat. PMID- 9745033 TI - Evolutionary conservation and tissue-specific processing of Hoxa 11 antisense transcripts. AB - We previously described the existence of abundant, processed, polyadenylated murine Hoxa 11 antisense transcripts. Of particular interest, in the developing limbs the antisense transcripts were observed to be present in a pattern complementary to that of the sense transcripts, suggesting a possible regulatory function (Hsieh-Li et al. 1995). We have analyzed the human HOXA 11 genomic locus, showing strong evolutionary conservation of regions potentially encoding antisense transcripts. Human HOXA 11 fetal kidney antisense cDNAs were identified and sequenced, demonstrating the evolutionary conservation of Hoxa 11 antisense transcription. As for the mouse, the human antisense RNAs were polyadenylated and showed several alternative processing patterns, but shared the sequences of a common 3' exon. The evolutionary conservation of the opposite strand transcripts strongly suggests function. A significantly long open reading frame was observed, but mouse-human comparisons argued against true coding function. Murine kidney Hoxa 11 antisense transcription and processing was also examined, revealing tissue-specific differences between limb and kidney. A novel procedure, designated Race in Circles, was devised and used to define mouse limb antisense transcription start sites. Furthermore, comparisons of human, mouse, and chicken sense transcript Hoxa 11 homeobox nucleotide sequences and their respective encoded homeodomains indicate a very strong selective pressure in vertebrates against mutations that result in coding changes. Given the significant differences in amino acid sequences of the homeodomains of different Hox genes, this observation argues for individual homeodomain functional specificity. PMID- 9745034 TI - Refined radiation hybrid map of mouse chromosome 17. AB - We have made a radiation hybrid map of mouse Chromosome (Chr) 17 with 75 microsatellite markers, including those from McCarthy et al. (Genome Res 7, 1153 1161, 1997). Seventy-four of the markers are linked at LOD > 9, and all link at LOD > 5. A LOD 3 framework of 18 markers was used to construct a placement map. The order obtained is in good agreement with genetic maps, and distance estimates give an idea of how recombination rates vary across the chromosome. Recombination is remarkably low with respect to RH break frequency in the region from the centromere to the end of H2. This is similar in interspecific and intersubspecific crosses despite the inversion of a substantial part of this region in Mus spretus with respect to Mus musculus. PMID- 9745035 TI - Sex-restricted non-Mendelian inheritance of mouse chromosome 11 in the offspring of crosses between C57BL/6J and (C57BL/6J x DBA/2J)F1 mice. AB - We report on the observation of sex-restricted, non-Mendelian inheritance over a region of mouse Chromosome (Chr) 11, occurring in the offspring of crosses between two commonly used Mus musculus-derived inbred strains, C57BL/6J and DBA/2J. In the surviving backcross progeny of reciprocal matings between (C57BL/6J x DBA/2J)F1 hybrids and the C57BL/6J parental strain, we observed the preferential appearance of C57BL/6J alleles along a region of Chr 11. The deviation from Mendelian predictions was observed only in female offspring from both reciprocal backcrosses, and not in males from either cross. The sex specificity of the observed non-Mendelian inheritance points to an explanation based on embryonic or neonatal lethality. Our data add to previously obtained evidence for a Chr 11 locus or loci with sex-specific and allele-specific effects on viability. PMID- 9745036 TI - Linkage mapping of fifty-eight new rat microsatellite markers. AB - Fifty-eight new anonymous simple sequence repeats (SSR) were generated and mapped to various rat chromosomes. Among them two genes (rat homologs for human cadherin 14 and mouse fibroblast growth factor-related protein) were mapped on Chromosomes (Chrs) 2 and 11 respectively. The majority of markers were generated from a small insert genomic library specific to Chr 11, 13, 14, and 15. Twenty new markers were mapped to Chr 13, which is known to contain a blood pressure quantitative trait locus (QTL). Several approaches to obtain microsatellite markers are described. The protocols and newly generated markers should be useful for ongoing rat genome project. PMID- 9745037 TI - Structure of the murine arginase II gene. AB - Mammals contain two genes encoding distinct isoforms of arginase (arginases I and II), both of which catalyze the conversion of arginine to ornithine and urea. However, their subcellular localization and tissue-specific patterns of expression are very different, indicating that they perform distinct physiologic roles. As an initial step in elucidating the regulation and physiologic roles of arginase II, this report describes the characterization of a mammalian arginase II gene. The murine arginase II gene contains eight exons like the arginase I gene. The six internal exons have intron/exon boundaries that are identical to the arginase I gene; however, exon three of the arginase II gene has obtained a three-base-pair insertion. The identity of the exon/intron boundaries is consistent with a gene duplication as the origin of the arginase isozymes with the small insertion occurring after the duplicative event. The promoter region of the arginase II gene, which bears no resemblance to that of the arginase I genes, contains numerous potential binding sites for enhancer and promoter elements but does not contain a TATA box. PMID- 9745038 TI - High-resolution mapping of sperm function defects in the t complex fourth inversion. AB - Structural variants of the mouse Chr 17-specific t complex, known as t haplotypes, express factors that alter the ability of sperm to carry out their roles in the normal fertilization process. In previous studies of males carrying heterospecific combinations of the t complex, we discovered a unique M. spretus/t haplotype phenotype of male sterility. In additional studies with mice carrying a series of M. spretus-M. m. domesticus recombinant Chr 17 homologs and a complete t haplotype (S-+/t), we monitored physiological aspects of sperm function to map a locus (Hst6) responsible for expression of the t-specific "curlicue" sperm flagellar curvature phenotype to 1 cM within the fourth inversion of the t complex. In the present report, we quantitatively analyze the in vitro capability of sperm from mice with similar S-+/t Chr 17 genotypes to fertilize zona pellucida-free mouse eggs. The results identify a locus, Stop1, mapping distal to Pim1, with acute effects on the ability of sperm to penetrate the oolemma. The data suggest that Stop1 is a complex locus consisting of at least two genetic elements, a proximal one overlapping the Hst6 locus, and another, distal to the Hst6 locus. Further quantitative analyses of the "curlicue" phenotype produced by sperm derived from these same animals indicate that expression of this chronic flagellar curvature phenotype also derives from at least two elements, both mapping within the Hst6 locus. Thus, these studies provide higher resolution mapping of the molecular basis of t haplotype-specific sperm dysfunction emanating from In(17)4. PMID- 9745039 TI - Genomic imprinting in ruminants: allele-specific gene expression in parthenogenetic sheep. AB - Studies in the mouse have established that both parental genomes are essential for normal embryonic development. Parthenogenetic mouse embryos (which have two maternal genomes and no paternal genome), for example, are growth-retarded and die at early postimplantation stages. The distinct maternal and paternal contributions are mediated by genomic imprinting, an epigenetic mechanism by which the expression of certain genes is dependent on whether they are inherited from mother or father. Although comparative studies have established that many imprinted mouse (and rat) genes are allele-specifically expressed in humans as well (and vice versa), so far imprinting studies have not been performed in other mammalian species. When considering evolutionary theories of genomic imprinting, it would be important to know how widely it is conserved among placental mammals. We have investigated its conservation in a bovid ruminant, the domestic sheep, by comparing parthenogenetic and normal control embryos. Our study establishes that, like in the mouse, parthenogenetic development in sheep is associated with growth retardation and does not proceed beyond early fetal stages. These developmental abnormalities are most likely caused by imprinted genes. We demonstrate that, indeed, like in mice and humans, the growth-related PEG1/MEST and Insulin-like Growth Factor 2 (IGF2) genes are expressed from the paternal chromosome in sheep. These observations suggest that genomic imprinting is conserved in a third, evolutionarily rather diverged group of placental mammals, the ruminants. PMID- 9745040 TI - A refined physical and EST map spanning 7.4 Mb of 10q24, a region involved in neurological disorders. PMID- 9745041 TI - Characterization of the mouse TGFbeta-inducible early gene (TIEG): conservation of exon and transcriptional regulatory sequences with evidence of additional transcripts. PMID- 9745042 TI - High-resolution mapping of the truncate (tc) locus on mouse chromosome 6. PMID- 9745043 TI - The mouse Lsp1 and Tnnt3 genes are 4.3 kb apart on distal mouse chromosome 7. PMID- 9745044 TI - Genomic organization of human B-ATF, a target for regulation by EBV and HTLV-1. PMID- 9745045 TI - Comparative mapping of the prion gene (PRNP) locus in cattle, sheep and human with PCR-generated probes. PMID- 9745046 TI - Eukaryotic DNA polymerases in DNA replication and DNA repair. AB - DNA polymerases carry out a large variety of synthetic transactions during DNA replication, DNA recombination and DNA repair. Substrates for DNA polymerases vary from single nucleotide gaps to kilobase size gaps and from relatively simple gapped structures to complex replication forks in which two strands need to be replicated simultaneously. Consequently, one would expect the cell to have developed a well-defined set of DNA polymerases with each one uniquely adapted for a specific pathway. And to some degree this turns out to be the case. However, in addition we seem to find a large degree of cross-functionality of DNA polymerases in these different pathways. DNA polymerase alpha is almost exclusively required for the initiation of DNA replication and the priming of Okazaki fragments during elongation. In most organisms no specific repair role beyond that of checkpoint control has been assigned to this enzyme. DNA polymerase delta functions as a dimer and, therefore, may be responsible for both leading and lagging strand DNA replication. In addition, this enzyme is required for mismatch repair and, together with DNA polymerase zeta, for mutagenesis. The function of DNA polymerase epsilon in DNA replication may be restricted to that of Okazaki fragment maturation. In contrast, either polymerase delta or epsilon suffices for the repair of UV-induced damage. The role of DNA polymerase beta in base-excision repair is well established for mammalian systems, but in yeast, DNA polymerase delta appears to fulfill that function. PMID- 9745047 TI - Amino-terminal sequences that direct nucleoporin nup153 to the inner surface of the nuclear envelope. AB - Nup153 is a large O-linked glycoprotein that is a component of the basket-like structure that forms the nucleoplasmic face of nuclear pore complexes (NPCs). The Nup153 molecule has a tripartite structure consisting of N- and C-terminal domains flanking a central zinc finger domain. All of the targeting and assembly information contained within Nup153 is contributed by the N-domain. In fact this region of the molecule can target a cytosolic protein, pyruvate kinase, to the nucleoplasmic face of the NPC. The zinc finger and C-terminal domains appear to have no role in these targeting and assembly activities. Deletion analysis reveals that there are two distinct regions within the Nup153 N-domain that contain different targeting functions. One of these is directly involved in assembly into the NPC while a second overlapping region may target Nup153, as well as other reporter molecules, to the inner face of the nuclear envelope. PMID- 9745048 TI - Direct visualisation of individual gene organisation in Trypanosoma brucei by high-resolution in situ hybridisation. AB - We achieved the direct visualisation of gene organisation in Trypanosoma brucei using fluorescent in situ hybridisation on extended nuclear DNA fibres. We demonstrated the repetitive nature of the tubulin gene cluster, which consists of up to 19 alpha- and beta-tubulin genes arranged in tandem repeats. PMID- 9745049 TI - Evolution of chromosome Y in primates. AB - We have investigated, by fluorescence in situ hybridization (FISH), the cytogenetic evolution of the Y chromosome in primates using 17 yeast artificial chromosomes, representative of the Y-specific euchromatic region of the human chromosome Y. The FISH experiments were performed on great apes (Homo sapiens, Pan troglodytes, Gorilla gorilla and Pongo pygmaeus pygmaeus), and on two Old World monkeys species as an outgroup (Cercopitecidae Macaca fascicularis and Papio anubis). The results showed that this peculiar chromosome has undergone rapid and unconstrained evolution both in sequence content and organization. PMID- 9745050 TI - Meiotic pairing and segregation of translocation quadrivalents in yeast. AB - Meiotic pairing and segregation were studied in three different heterozygous reciprocal translocation strains of the baker's yeast, Saccharomyces cerevisiae. Pachytene translocation quadrivalents were identified by a combination of immunofluorescence and fluorescence in situ hybridization and the karyotypes of meiotic products were determined by pulsed-field gel electrophoresis. The translocations differed with respect to the relative sizes of the chromosomes involved and the positions of translocation breakpoints, and produced translocation quadrivalents of widely different shapes. This allowed us to study the influence of the morphology of quadrivalents on their segregation behaviour. In all cases alternate predominated over adjacent segregation. 3:1 disjunction of chromosomes was more frequent when translocation breakpoints were close to the centromeres. If a translocation breakpoint was distant from the centromere, the occurrence of an intervening chiasma influenced the pattern of segregation. In general, quadrivalent formation and segregation resembled the behaviour of translocation heterozygotes in most higher eukaryotes. We therefore conclude that, although chromosome condensation does not occur in yeast metaphase, centromere orientation and chromosome disjunction are governed in a way similar to that of higher eukaryotes. PMID- 9745051 TI - Differential stability of a human mini-chromosome in mouse cell lines. AB - A 4 Mb human mini-chromosome, DeltaDelta2, was transferred from Chinese hamster ovary (CHO) cells into a mouse L cell line. The mini-chromosome could be transferred intact into the L cells, with 112/119 clones maintaining a mini chromosome of the same size as the original. Ten clones were grown for 30 days in continuous culture. The mini-chromosomes were maintained stably with or without selection at a copy number of 1-2 per cell and none experienced any size alterations, as determined by pulsed-field gel electrophoresis. Thus DeltaDelta2 is structurally and mitotically stable in L cells. This contrasts with results in embryonic stem cells, in which DeltaDelta2 is highly unstable. These findings indicate that established somatic cell lines, such as L cells and CHO cells, have less stringent controls over centromeric function than do normal embryonic cells. PMID- 9745052 TI - CpG islands detected by self-primed in situ labeling (SPRINS). AB - We have studied the distribution and methylation of CpG islands on human chromosomes, using the novel technique of self-primed in situ labeling (SPRINS). The SPRINS technique is a hybrid of the two techniques primed in situ labeling (PRINS) and nick translation in situ. SPRINS detects chromosomal DNA breaks, as in nick translation in situ, and not annealed primers, as is the case in PRINS. We analyzed in situ-generated DNA breaks induced by the restriction enzymes HpaII and MspI. These restriction enzymes enable the detection of chromosomal CpG islands. Both HpaII- and MspI-SPRINS produce a banding pattern resembling R banding, indicating a higher level of CpG islands in R-positive bands than in R negative bands. Our SPRINS banding observations also indicate differences in sequence copy number in the satellites of homologous acrocentric chromosomes. Furthermore, a comparison of homologous HpaII-SPRINS-banded X chromosomes of females from lymphocyte cultures grown without methotrexate or bromodeoxyuridine revealed methylation difference between them. The same comparison of homologous X chromosomes from the cell line GM01202D, which has four X chromosomes, one active and three inactive, revealed the active X chromosome to be hypermethylated. PMID- 9745053 TI - Distribution of heterochromatin on the mitotic chromosomes of Musca domestica L. in relation to the activity of male-determining factors. AB - In the housefly, male sex is determined by a dominant factor, M, located either on the Y, on the X, or on any of the five autosomes. M factors on autosome I and on fragments of the Y chromosome show incomplete expressivity, whereas M factors on the other autosomes are fully expressive. To test whether these differences might be caused by heterochromatin-dependent position effects, we studied the distribution of heterochromatin on the mitotic chromosomes by C-banding and by fluorescence in situ hybridization of DNA fragments amplified from microdissected mitotic chromosomes. Our results show a correlation between the chromosomal position of M and the strength of its male-determining activity: weakly masculinizing M factors are exclusively located on chromosomes with extensive heterochromatic regions, i.e., on autosome I and on the Y chromosome. The Y is known to contain at least two copies of the M factor, which ensures a strong masculinizing effect despite the heterochromatic environment. The heterochromatic regions of the sex chromosomes consist of repetitive sequences that are unique to the X and the Y, whereas their euchromatic parts contain sequences that are ubiquitously found in the euchromatin of all chromosomes of the complement. PMID- 9745054 TI - Assignment of linkage groups to pea chromosomes after karyotyping and gene mapping by fluorescent in situ hybridization. AB - Chromosomes of the pea (Pisum sativum L.) were submitted to fluorescent in situ hybridization (FISH) with probes specific for the oligonucleotides (AG)12, (AC)12, (GAA)10, and (GATA)7 and for the genes encoding 25S rRNA, 5S rRNA and the storage proteins legumin A, K and vicilin. A fourth 5S rRNA gene locus, apparently specific for an accession of the cultivar Grune Victoria, was newly detected. This allowed all seven chromosome pairs to be distinguished by FISH signals of rRNA genes. The same was possible using a combination of oligonucleotide probes or of oligonucleotides and rRNA gene-specific probes in multicolour FISH. Rehybridization with the 5S rRNA gene-specific probe allowed us to assign vicilin genes to the short arm of chromosome 5, the single legumin A locus to the long arm of chromosome 3 and the legumin B-type genes (exemplified by legumin K) to one locus on the short arm of chromosome 6. Correlation of these data with an updated version of the pea genetic map allowed the assignment of most linkage groups to defined chromosomes. It only remains to be established which of linkage groups IV and VII corresponds to the satellited chromosomes 4 or 7, respectively. PMID- 9745055 TI - Cost efficacy of laparoscopic vs open surgery. Hospitals vs community. PMID- 9745056 TI - A cost and outcome comparison between laparoscopic and Lichtenstein hernia operations in a day-case unit. A randomized prospective study. AB - BACKGROUND: Laparoscopic hernia repair has often been criticized for its high costs. METHODS: To compare the costs of laparoscopic and open hernia repair, 40 patients were randomized for either transabdominal laparoscopic or Lichtenstein mesh repair (under local anesthesia) in a day-case surgery unit. RESULTS: Median operative times for the laparoscopic and open groups were 62 and 65 min, respectively. Postoperative pain was comparable for the two groups. The period before return to normal life was 14 days in the laparoscopic group and 21 days in the open group. The hospital costs were 2051 FIM ($1 US = 4.6 FIM) higher in the laparoscopic group, but the total costs for employed patients (including expenses due to lost work days) were lower. CONCLUSION: Although the Lichtenstein operation is cheaper for the hospital, the total costs for working patients are lower with the laparoscopic technique, when the cost of lost work days is factored into overall expense. PMID- 9745057 TI - Cost-effective appendectomy. Open or laparoscopic? A prospective randomized study. AB - BACKGROUND: The aim of this study was to compare the outcome and cost effectiveness of laparoscopic (LA) and open appendectomy (OA). METHODS: Forty consecutive patients were randomized to either the LA (n = 19) or OA (n = 21) group. RESULTS: The medians of operative times in the LA and OA groups were 31.5 and 41 min, respectively. The total operation room times were 91 and 82 min, respectively. There was no significant difference in postoperative pain or fatigue, but return to normal life was faster in the LA group (14 versus 26. 5 days). The median hospital costs per patient were 8,538 and 6,788 FIM ($1 US = 4.6 FIM) in the LA and OA groups, respectively; but the total costs among working patients were lower in the LA group (20, 963 versus 27,778 FIM) due to faster return to work. CONCLUSIONS: Laparoscopic appendectomy is as safe as open appendectomy. The hospital costs are higher, but LA offers significant cost savings to the payer for working patients. PMID- 9745058 TI - Laparoscopic or open fundoplication? A complete cost analysis. AB - BACKGROUND: As part of a prospective observational trial, we set out to determine the direct and indirect costs of an open versus a laparoscopic fundoplication for chronic gastroesophageal reflux disease (GERD). METHODS: Two groups of patients, each comprising 28 subjects, were studied. RESULTS: All patients received a functioning fundoplication that did not require any additional therapy. Because 19 and 12 patients in the open and laparoscopy groups, respectively, were employed in the work force, we were able to assess the costs due to loss of production. The mean operating time was similar for both groups, but postoperative stay differed significantly; though it amounted to 8 days for the open group, it was only 2 days for the laparoscopy group. Postoperative sick leave was 29.9 days in the open and 9.9 in the laparoscopy group (p < 0.05). The costs of the operations were 18,363 SEK for laparoscopy and 12,856 SEK for conventional fundoplication. On the other hand, the cost for hospital stay amounted to 35,488 SEK in the open group but was only 25,571 SEK for those undergoing laparoscopy. When we add outpatient visits, endoscopies, and other medical expenses, the total direct costs in the laparoscopy group come to 27,693 SEK, as compared to 37,482 SEK for the open fundoplication. The indirect medical costs, which were dominated by loss of production (36,732 versus 12,126 SEK), came to 37,126 and 12,595 SEK in the open and laparoscopy groups, respectively. CONCLUSIONS: The total community-based costs for the open and laparoscopic operations for chronic GERD amounted to 74,608 and 40,289 SEK, respectively. Thus, we would recommend the laparoscopic procedure in most cases. PMID- 9745059 TI - Femoral venous flow dynamics during intraperitoneal and preperitoneal laparoscopic insufflation. AB - BACKGROUND: Laparoscopic herniorrhaphy may be performed using an intraperitoneal or a preperitoneal approach. Anecdotal and experimental evidence indicates that alterations in lower extremity venous flow, which occur during intraperitoneal laparoscopic insufflation, may be associated with an increased risk of deep vein thrombosis. However, no study has directly compared femoral venous flow during intraperitoneal insufflation with that during preperitoneal insufflation. METHOD: In eight consecutive patients undergoing laparoscopic herniorrhaphy under general anesthesia, flow through the common femoral vein was evaluated with B-mode and color flow duplex. Pre- and intraperitoneal pressures were standardized to 10 mm Hg, and respiratory tidal volumes were standardized to 10 cc/kg. Flow measurements were taken at end expiration. Flow through the common femoral vein was measured after induction of anesthesia, during intraperitoneal insufflation, during preperitoneal insufflation, and between insufflations to ensure return to baseline. RESULTS: All patients in the study were males. Their mean age was 59 years. Mean flow in the common femoral vein was essentially identical at baseline (138 ml/min) and during preperitoneal insufflation (135 ml/min). Alternatively, mean flow in the common femoral vein was significantly reduced during intraperitoneal insufflation (65 ml/min, p = 0.02). CONCLUSIONS: Flow in the common femoral vein is significantly reduced during intraperitoneal insufflation. However, flow in the common femoral vein is not affected by preperitoneal insufflation. These data suggest that laparoscopic preperitoneal inguinal hernia repair may pose as less a risk of thromboembolic complications than laparoscopic intraperitoneal inguinal hernia repair. PMID- 9745060 TI - Lack of neurohumoral response to pneumoperitoneum for laparoscopic cholecystectomy. AB - BACKGROUND: Pneumoperitoneum (PP) for laparoscopic surgery induces prompt changes in circulatory parameters. The rapid onset of these changes suggests a reflex origin, and the present study was undertaken to evaluate whether release of vasopressor substances could be responsible for these alterations. The influence of two different anesthesia techniques was also evaluated. METHODS: American Society of Anesthesiologists (ASA) class I patients, scheduled for laparoscopic cholecystectomy, were investigated. The first group (n = 10) was anesthetized intravenously. The second group (n = 6) had inhalation anesthesia. Plasma vasopressin, catecholamines, and plasma renin activity were investigated as neurohumoral vasopressor markers of circulatory stress. The general stress response to surgery was assessed by analysis of plasma cortisol. RESULTS: Induction of pneumoperitoneum caused no apparent activation of vasopressor substances, although several hemodynamic parameters responded promptly. CONCLUSION: The hemodynamic alterations, seen at the establishment of PP during stable anesthesia, cannot be explained by elevation of vasopressor substances in circulating blood. PMID- 9745061 TI - Pneumoperitoneum risk prognosis and correction of venous circulation disturbances in laparoscopic surgery. A pilot study. AB - BACKGROUND: This study was initiated to find a method of determining the prognosis for possible changes in hemodynamic and respiratory parameters in patients with pneumoperitoneum (PP). METHODS: We devised a model for a pseudopneumoperitoneum (PPP), which is created by encircling the wide pneumochamber on the entire abdomen and inflating it to a preset pressure. To verify the prognostic possibilities of the proposed model, we studied the pneumotachygraphy parameters, noninvasive and invasive monitoring parameters of PPP after induction of anaesthesia, and venous circulation disturbances, as well as the medical effect of the intermittent sequential compression device. RESULTS: In healthy patients, the restrictive lung syndrome did not approach the risky limit. In patients >/=60 years old, this syndrome was very close to the limit. In a number of patients with serious cardiovascular and pulmonary pathology, the pressure of >10 mmHg was considered to be intolerable. Lung compliance, which was the parameter most sensitive to the increased intraabdominal pressure, was 47 +/- 10 at baseline, and 29 +/- 4 (p > 0.05) at both PPP and real PP (14 mmHg). CONCLUSIONS: The PPP model is quite similar to the real PP and can be used for preoperative prognosis in laparoscopic surgery. The elevated intraabdominal pressure results in a significant disturbance of venous blood flow in the lower extremities. The use of the device for peristaltic pneumomassage of the lower limbs is effective in correcting negative changes in venous hemodynamics in laparoscopic surgery. PMID- 9745062 TI - Can sonographic signs predict conversion of laparoscopic to open cholecystectomy? AB - BACKGROUND: The aim of this study was to determine whether sonographic signs can predict the risk for conversion of laparoscopic (LC) to open cholecystectomy (OC). METHODS: All 346 patients who underwent LC at our institution between January 1, 1993, and March 1, 1996, were studied retrospectively. Patients who had no sonographic examination during 6 months prior to surgery and patients treated by inexperienced surgeons were excluded from the study. Patient characteristics and sonographic parameters were evaluated by univariate and multivariate analysis, using conversion to OC as a dependent variable. RESULTS: In 23 of 134 patients (17.2%), LC was converted to OC. In the univariate analysis, gallbladder distention (>4.5 cm; relative risk [RR] 3.5; 95% confidence intervals [CI] 1.7-5.3), stone impaction (RR 2.4; 95% CI 1.1-5.1), thickened gallbladder wall (RR 2.4; 95% CI 1.2-5.1), and acute cholecystitis (RR 2.6; 95% CI 1.1-6.7) were able to predict the need for conversion. Logistic regression defined only the sonographic sign of distention of the gallbladder as a predictor of conversion. CONCLUSIONS: Gallbladder distention as a sonographic sign is associated with a high relative risk for conversion. The predictive value of sonographic signs for conversion requires further assessment in a prospective study. PMID- 9745063 TI - Is laparoscopic sonography a reliable and sensitive procedure for staging colorectal cancer? A comparative study. AB - BACKGROUND: Laparoscopic colectomy has developed rapidly with the explosion of technology. In most cases, laparoscopic resection is performed for colorectal cancer. Intraoperative staging during laparoscopic procedure is limited. Laparoscopic ultrasonography (LUS) represents the only real alternative to manual palpation during laparoscopic surgery. METHODS: We evaluated the diagnostic accuracy of LUS in comparison with preoperative staging and laparoscopy in 33 patients with colorectal cancer. Preoperative staging included abdominal US, CT, and endoscopic US (for rectal cancer). Laparoscopy and LUS were performed in all cases. Pre- and intraoperative staging were related to definitive histology. Staging was done according to the TNM classification. RESULTS: LUS obtained good results in the evaluation of hepatic metastases, with a sensitivity of 100% versus 62.5% and 75% by preoperative diagnostic means and laparoscopy, respectively. Nodal metastases were diagnosed with a sensitivity of 94% versus 18% with preoperative staging and 6% with laparoscopy, but the method had a low specificity (53%). The therapeutic program was changed thanks to laparoscopy and LUS in 11 cases (33%). In four cases (12%), the planned therapeutic approach was changed after LUS alone. CONCLUSIONS: The results obtained in this study demonstrate that LUS is an accurate and highly sensitive procedure in staging colorectal cancer, providing a useful and reliable diagnostic tool complementary to laparoscopy. PMID- 9745064 TI - Management of Mirizzi's syndrome in the laparoscopic era. AB - BACKGROUND: Mirizzi's syndrome is an uncommon cause of common hepatic duct obstruction resulting from gallstone impaction in the cystic duct or gallbladder neck. The role of laparoscopic surgery in the treatment of this condition is still not well defined. This article reports six cases of Mirizzi's syndrome and comments on the management of this condition using the laparoscopic approach. METHODS: A review of 878 consecutive cholecystectomies from July 1991 to July 1996 identified six cases of Mirizzi's syndrome (0.7%) that were approached laparoscopically. RESULTS: This study involved three men and three women with mean age of 64 (range, 57-70) years. All cases were approached by laparoscopy. One case was converted because of unclear anatomy in the Calot's triangle due to dense adhesions; open cholecystectomy, exploration of the common bile duct and T tube insertion was performed. The other five cases were successfully managed laparoscopically. Subtotal cholecystectomy was performed in two cases, and in three patients with cholecystocholedochal fistula, the defect was closed over a T tube. There was no postoperative morbidity or mortality. A follow-up period of 8 to 17 (mean, 12) months revealed no complications. CONCLUSIONS: Laparoscopic management of Mirizzi's syndrome is feasible and safe but can be technically demanding. A policy of trial dissection by an experienced laparoscopic surgeon is recommended, and if anatomy remains unclear, it is prudent to convert. PMID- 9745065 TI - Aseptic laparoscopic colon resection with intraabdominal anastomosis. An experimental study in pigs. AB - BACKGROUND: We evaluated a new aseptic method for laparoscopic left colon resection in terms of technical feasibility and outcome. METHODS: Ten pigs were operated on under general anesthesia. Pre- and postoperative body weight, stools, behavior, and need for analgesics were recorded. Fourteen days later, the animals were killed. At autopsy, the degree of intraabdominal adhesions was noted. The anastomoses were sent for histological examination. The entire procedure was performed intracorporeally, and no antibiotics were given. After division of the mesocolon, the segment to be resected was invaginated down through the colon. This was facilitated by a custom-made instrument that was introduced into the bowel via the anus; it consisted of a pull-out device and a modified diathermy wire. The anastomosis was completed at the invagination fold by a row of hernia staples that were covered by an interrupted suture. Then the invaginated bowel was transected by the diathermy wire and delivered through the anus. RESULTS: One animal was killed before completion of the operation because of a colonic perforation. The remaining nine animals had an uneventful and rapid recovery. They ate from the 1st postoperative day and gained weight rapidly. Stools were normal after 2 days (median), and normal behaviour was noted in all animals from the 1st postoperative day. At the postmortem examination, intraabdominal adhesions were observed in two animals. In one case, the adhesions extended from a hematoma in the mesentery to the abdominal wall. There were no adhesions to the anastomosis or the colon. In the other case, the anastomosis adhered to the right uterine tube and a loop of small intestines. CONCLUSIONS: The method is technically feasible, but a modification is suggested for cases where the invagination is impossible. Recovery after the operation is rapid. PMID- 9745066 TI - Percutaneous drainage of pancreatic pseudocyst into the stomach. AB - BACKGROUND: We present our experience with percutaneous ultrasonographically guided internal cystogastric drainage of pancreatic pseudocysts using a double pigtail catheter. METHODS: In nine patients, the pancreatic pseudocysts following acute pancreatitis were drained percutaneously into the stomach with the double pigtail catheter under ultrasonographical (US) control. The needle insertion through both gastric walls and the final position of the proximal curve of the catheter were monitored with a gastroscope. The position of the distal curve of the catheter was checked by US. There were no procedure-related complications. The patients were followed up monthly by clinical and US examination. RESULTS: At first follow-up 1 month after the intervention, none of the patients had evidence of the pseudocyst. The patients were not aware of the catheter and functioned normally throughout the procedure and catheter removal. The catheter was removed endoscopically after 5-8 months. CONCLUSIONS: The method is minimally invasive and also feasible in high-risk surgical patients. It requires a team consisting of an interventional radiologist, an ultrasonographer, and an endoscopist. In properly selected patients, the results are excellent. PMID- 9745067 TI - Thoracoscopic transdiaphragmatic microwave coagulation therapy for a liver tumor. AB - BACKGROUND: Microwave coagulation therapy (MCT) for hepatocellular carcinoma, which induces tumor coagulonecrosis, is now recognized as an efficient treatment. However, when a tumor is located just below the top of the diaphragmatic dome, laparotomical MCT requires a large incision, and percutaneous MCT is sometimes impossible. PATIENTS AND METHODS: The patients were four men and two women. There were four cases of hepatocellular carcinoma and two cases of liver metastasis from colorectal cancer. All tumors were located below the top of the diaphragmatic dome. Thoracoscopic transdiaphragmatic MCT (TTMCT) was performed under general anesthesia using an endotracheal double-lumen tube. Identification of the tumor site in the liver was performed using an ultrasonic probe under thoracoscopic observation. After the diaphragm above the tumor was opened, a needle electrode to transmit microwaves was inserted directly into the tumor. Microwave irradiation was repeated to coagulate the entire lesion. After completion of TTMCT, the diaphragm was closed thoracoscopically. RESULTS: TTMCT was successfully administered to cancerous lesions in all six patients. The postoperative course was uneventful, and the average postoperative hospitalization period was 10.5 days. None of the patients has shown any recurrence during a follow-up period of 4-23 months. CONCLUSIONS: TTMCT was performed without any difficulty using the thoracoscopic surgical technique, and its therapeutic outcome was satisfactory. This is effective for tumors located just below the top of the diaphragmatic dome. PMID- 9745068 TI - Sac excision is essential to adequate laparoscopic repair of paraesophageal hernia. AB - BACKGROUND: We compared the incidence of early hernia recurrence in nonrandomized but consecutive patients undergoing laparoscopic repair of paraesophageal hernia (LRPH) without and with excision of the hernia sac. METHODS: LRPH was completed in 55 of 58 patients. In the first 25 patients, the sac was not excised. Total sac excision was performed in the subsequent 30 patients. All patients had crural repair with or without fundoplication, or gastropexy. RESULTS: Mean age of patients was 68 years (range, 34-95). There were three conversions; one patient died postoperatively. Mean operative time was 225 min in the first group and 190 min in the sac excision group. Median length of stay was 2 days (range, 1-15) for both groups. CONCLUSIONS: A precise method of total sac excision simplified dissection. It also ensured complete reduction of the hernia and availability of adequate esophageal length. Operative time was not increased, and no subsequent early recurrences were observed (p < 0.05). PMID- 9745069 TI - Early experience with extraperitoneal endoscopic radical retropubic prostatectomy. AB - This article reports our early experience using laparoscopic instruments and techniques when performing radical retropubic prostatectomy through an entirely extraperitoneal endoscopic approach. Two patients with localized adenocarcinoma of the prostate underwent endoscopic radical retropubic prostatectomy through an entirely extraperitoneal approach (EERRP). The procedure was evaluated for its efficacy in removing prostate and seminal vesicles and in effecting complete vesicourethral anastomosis. Operative time, blood loss, hospital stay, and pathology were also evaluated. Complete endoscopic removal of the prostate and seminal vesicles was achieved in both patients. Endoscopic reconstruction of the bladder neck with watertight anastamosis was successful in both. Operative time and estimated blood loss improved from 5 h and 45 min and 600 cc, respectively, in patient 1 to 4 h and 400 cc in patient 2. Hospital stay was 2.5 days for both. The early experience for EERRP is encouraging. Further evaluation to standardize technique and determine its efficacy and role in treating prostate cancer is in order. PMID- 9745070 TI - Laparoscopic treatment of congenital choledochal cyst. AB - We describe the laparoscopic treatment of a patient presenting with congenital choledochal cyst. Our patient was a 19-year-old man with a complaint of recurrent abdominal pain due to pancreatitis. The choledochal cyst was type I and had a common channel of pancreatobiliary duct, as revealed by endoscopic retrograde cholangiopancreatography. Under laparoscopic guidance, the dilated bile duct and the gallbladder were excised, and a Roux-en-Y anastomosis was constructed with an endo-EEA. Finally, end-to-side anastomosis was carried out by the continuous suture method, aided by an Endostitch between the stump of the hepatic duct and the Roux-en-Y limb. After the operation, slight hyperamylasemia was observed for several days but further treatment was not necessary. Postoperative symptoms were minimal, and the patient was discharged on the 11th day after the procedure. Although it is difficult and time-consuming, laparoscopic operation is highly beneficial for the patient. The use of such instruments as the endostapler and Endostitch may help to simplify this complex intracorporeal procedure involving division and anastomosis of the digestive tract. PMID- 9745071 TI - Laparoscopic management of a large ovarian cyst in the neonate. AB - Laparotomy has become the preferred approach to the excision of large, complex abdominal cysts in the neonate. We describe a laparoscopic-assisted decapsulation of an antenatally diagnosed abdominal cyst that was noted on postnatal ultrasound scan to have a complex echo pattern. This limited procedure allows for accurate verification of the diagnosis, institution of appropriate therapy, and organ salvage. It represents a superior management option that obviates the significant complications associated with conservative management. PMID- 9745072 TI - Sterile and economic instrumentation in laparoscopic surgery. Experiences with 6,000 surgical laparoscopies, 1990-1996. AB - BACKGROUND: Because so many common surgical problems can now be addressed by the laparoscopic approach, the issue of sterile processing has to be reconsidered. METHODS: Selected laparoscopic instrumentation was analyzed regarding wear and tear and decontamination after sterile processing following 6,000 surgical laparoscopies carried out between 1990 and 1996 at the Academic Hospital Moabit, Berlin. RESULTS: Fewer than 7.9 (parts of) instruments failed per 100 laparoscopies. Most of the repairs involved scissors. The main problems were blunting, burnt or disconnected electromechanical components, defective insulation, and damaged or lost parts of dismantable instruments. Residues of human blood proteins were detected on a few instruments. The effect of intraluminal rinsing was documented by measuring the iron content (as an indicator for blood contamination). A comparison of costs showed that it was >10 times cheaper to use instrumentation with reusable components. CONCLUSIONS: The sterile processing of economic reusable instrumentation for laparoscopies needs staff well trained in sterile supply. Instrument design should allow easy dismantling and rinsing of internal parts. Insulating compounds present a problem for decontamination. Disinfection with aldehydes before cleaning the lumina of instruments must be avoided because protein coagulation will occur. A tube-in tube concept for tubular instruments offering compatibility should be favored. PMID- 9745073 TI - Bile duct injuries during laparoscopic cholecystectomy. PMID- 9745074 TI - Mesh repair of paracolostomal hernia by laparoscopy. PMID- 9745075 TI - News and notices PMID- 9745076 TI - Staging systems for pancreatic cancer: differences between the Japanese and UICC systems. AB - Differences between the clinical staging system of the Japan Pancreas Society (JPS) and the Union Internationale Contre le Cancer (UICC) stage classification may account for reported differences in the prognosis of pancreatic carcinoma between Japan and the West. In the review, we compared the characteristics of the JPS and UICC staging in 1689 patients, registered with the JPS from 1981 to 1990, who underwent resection for carcinoma of the pancreatic head. The survival rates correlated well with the JPS stage classification. The UICC staging did not reflect differences in prognoses among the stages. The current JPS staging system, introduced in 1993, still differs from that of the UICC. To compare the results of treatment for patients with pancreatic cancer it is important to establish a more practical and universal staging system for carcinoma of the pancreas. PMID- 9745077 TI - Staging and treatment for patients with pancreatic cancer. How small is an early pancreatic cancer? AB - To determine the tumor size that constitutes early pancreatic cancer, we reviewed and analyzed the English-language and Japanese literature (a total of 25 publications) on small pancreatic cancers less than 2 cm in diameter and/or stage 1 cancers. Reports on in situ carcinoma and intraductal carcinoma of the pancreas were also evaluated. The results were: (1) A total of 302 cases of small pancreatic cancer less than 2 cm in diameter reported at separate institutions were pooled from 15 reports. The rates for patients in stage I and those with no lymph node metastasis averaged 41.7% and 57.9%, respectively. The 5-year postoperative cumulative survival rate (5Y-PCR) was less than 50% in almost all these reports. Similar data were shown in the 7 collective reviews. (2) Another 33 cases of small pancreatic cancer of 1 cm or less in diameter were collected from three reports. The rates for stage I tumor and 5Y-PCR at one institution with two reports were 100% and 100% and the rates in the other report were 85% and 78%, respectively. (3) Twelve cases of in situ carcinoma and intraductal carcinoma of the pancreas were collected from four reports. All of the patients were stage I and were alive with no evidence of tumor recurrence for periods ranging from 6 to 78 months. Small pancreatic cancer less than 1 cm in diameter is better viewed as an early pancreatic cancer, and in situ carcinoma and intraductal carcinoma of the pancreas with minimal invasion to the pancreatic parenchyma may be defined as early pancreatic cancer, regardless of size. PMID- 9745078 TI - Surgical treatment for patients with advanced pancreatic cancer. PMID- 9745079 TI - What are the problems associated with the surgical treatment of patients with pancreatic cancer? AB - Modern surgical therapy of pancreatic cancer has resulted in few long-term survivors. There are several reasons. First, most patients are not diagnosed in an early tumor stage, resulting in a minority of patients undergoing surgical resection. A breakthrough in screening methodology is required until this most major of obstacles can be overcome. Second, inaccurate clinical tumor staging is all that is available for the majority of patients, since most patients are not resected. Imaging techniques used for clinical staging require anatomic verification before clinical staging can be reliable. Then adequate comparison of treatments for patients who never receive anatomical staging can accomplished. Postoperative tumor staging using anatomical methods from intraoperative findings and examination of a surgical specimen require a staging system that is simple and directly correlates with survival. The best staging system can be developed only with international cooperation. An adequate comparison of the results of treatment will then be possible. Two broad treatment areas that are most promising are surgical (extending resections to yield negative surgical margins) and adjuvant protocols (beginning with a variety of radio sensitizing chemotherapeutic agents). PMID- 9745080 TI - Results of surgery and adjuvant radiotherapy for pancreatic cancer. AB - We retrospectively reviewed the cases of 34 patients with pancreatic cancer who underwent resection between January 1988 and December 1996. Adjuvant radiotherapy was performed in 24 patients, with 13 receiving both intra- and postoperative radiotherapy, 2 receiving postoperative radiotherapy (PORT) alone, and 9 receiving intraoperative radiotherapy (IORT) alone. The 1- and 3-year survival rates for all 34 patients were 59% and 19%, respectively, with a median survival of 13 months. At the time of the analysis, three patients were still alive. Recurrence patterns were assessed in 25 patients who had had no distant metastases at the time of surgery, had survived more than 3 months after surgery, and had undergone close surveillance for recurrence. Based on computed tomography (CT) and autopsy findings, a total of 15 (60%) of these 25 patients had local recurrence, 13 (52%) had liver metastases, and 8 (32%) had both. Eight (62%) of the 13 patients who received IORT and/or PORT developed local recurrence, and we failed to detect any survival advantage of IORT and/or PORT over surgery alone. However, autopsies revealed a suppressive effect of radiation on cancer growth, and local recurrence was not considered to be the direct cause of death in any of the patients, nor did any of the patients develop gastrointestinal obstruction due to local recurrence. The incidence of liver metastasis in the patients with and without tumor invasion of the portal system was 80% (8/10) and 33% (5/15), respectively. The patients who did not develop liver metastasis had significantly longer survivals than who did. Further improvements of survival await effective prophylactic treatment for liver metastases. PMID- 9745081 TI - Intraoperative radiotherapy for pancreatic adenocarcinoma. AB - Intraoperative radiotherapy (IORT) is an innovative treatment approach for cancer of the pancreas. The common causes of treatment failure in pancreatic cancer are regional recurrence and distant metastasis. While at present the benefit of IORT in unresectable pancreatic cancer is still controversial and awaits further prospective trials for its clarification, the experience gathered over a period of 30 years with IORT for pancreatic cancer does suggest that IORT should be part of the adjuvant therapy of surgical resection. A combination with pre- or postoperative external beam radiotherapy and chemotherapy may be beneficial for both resectable and unresectable patients. IORT was shown to be a relatively safe intervention and it notably improved the quality of life of patients with locally advanced pancreatic carcinomas by alleviating their pain. Here, we summarize and discuss the experience reported to date and present our historical analysis of IORT for pancreatic cancer. PMID- 9745082 TI - Primary malignancies of the hepato-biliary-pancreatic system in organ allograft recipients. AB - In a series of 10151 organ allograft recipients who developed 10813 de novo malignancies after transplantation, 755 involved the hepato-biliary-pancreatico duodenal (HBPD) area. If nonmelanoma skin cancers and in situ carcinomas of the uterine cervix were excluded (as they are from most cancer statistics), then the HBPD area was affected by 10% of neoplasms. Many of the tumors encountered were uncommon in the general population. The largest group of neoplasms was 474 lymphomas, which comprised 63% of the total. Other major malignancies were hepatocellular carcinomas (HCC; 15%), pancreatic carcinomas (11%), cholangiocarcinomas (3%), Kaposi's sarcomas (3%), and other sarcomas (1%). Lymphomas occurred at a younger age than other tumors (average, 39 versus 50 years), appeared earlier after transplantation (average, 24 versus 77 months), and were more frequently associated with immunosuppressive therapy with the antilymphocytic agents (ALG/ATG) and/or (OKT3) (59% versus 28%). Lymphomas were localized to the HBPD area in only 18% of patients, whereas in 82% there was involvement of other organs or sites. The liver was involved in 95% of lymphomas. Lymphomas frequently involved allografts, the liver in 84%, and the pancreas in 59%. Of 292 patients treated for lymphomas 67 (23%) had complete remissions lasting 6 months or more. HCC was frequently associated with hepatitis B or C infection. Kaposi's sarcomas were rarely confined to the HBPD area, and in 25% of cases there were no associated skin lesions. An unusual subset of tumors were leiomyosarcomas involving hepatic allografts of pediatric patients. The poor prognosis of most tumors in this series may be related to delays or problems in making the diagnosis in these immunosuppressed patients and, perhaps, it may also be related to the unusually aggressive behavior of some tumors. PMID- 9745083 TI - Basic knowledge of a microwave tissue coagulator and its clinical applications. AB - A microwave tissue coagulator originally developed to control hemorrhage during hepatic resection has recently found widespread use in the field of minimally invasive surgery. This surgical tool is based on the principle that by radiating a 2450-MHz (12-cm wavelength) microwave from a monopolar antenna within tissue, the heat generated will be limited to within the electromagnetic field generated around the antenna, leading to coagulation of protein in that field. It is therefore possible to use this monopolar antenna as a surgical electrode. The coagulation field is determined by the relationship between the wavelength frequency, tissue-specific permittivity, antenna length, waveform and output, and duration of the irradiation. Since this technique has been applied to devise a new method of hepatectomy, it has also found use in various other surgical fields, such as gastrointestinal tract endoscopic surgery, laparoscopic surgery, and percutaneous surgery. It has also enhanced therapeutic results, notably in cancer therapy. PMID- 9745084 TI - Systematic laparoscopic left lateral segmentectomy of the liver for hepatocellular carcinoma. AB - A systematic technique for the resection of hepatocellular carcinoma (HCC) prevents the dissemination of cancer cells through the portal vein of the remnant liver. We successfully performed a systematic laparoscopic left lateral segmentectomy in a 62-year-old man with HCC. The tumor was located in the left lateral segment of the liver, and measured approximately 4 cm in diameter. Since no other tumors were detected in the liver or in any distant organs, the patient was considered to be a candidate for surgery. A laparoscopic hepatic resection was selected as the procedure of choice. Prior to dissection of the liver parenchyma, the arteries and branches of the portal vein feeding the left lateral segment were divided and dissected, together with the branches of the biliary tree in the umbilical portion of the left pedicle of Glisson's capsule. The liver parenchyma was then dissected and the left hepatic vein divided and dissected, and transection of the left lateral segment was completed. The patient's postoperative course was uneventful and he was discharged on postoperative days 14. No evidence of recurrence has been noted in the 22 months after surgery (the time of this report). This less invasive surgery, taking into consideration the pathogenesis of HCC, may be a useful new approach in selected patients with this tumor. PMID- 9745085 TI - The management of hepatic hydatid cysts: review of 94 cases. AB - In this retrospective study, 94 patients operated for hepatic hydatid cysts were reviewed to compared the advantages and disadvantages of different operative techniques. The patients were divided into four groups according to the type of operation. Group I consisted of 33 patients with peripherally located small cysts, eligible for excision, who underwent cystectomy. Group II consisted of 28 patients with cysts smaller than 5 cm, not suitable for complete removal, who underwent partial cystectomy with capitonnage. Group III were 21 patients with cysts larger than or equal to 5 cm, not suitable for complete removal, who underwent partial cystectomy with omentoplasty. Infection and biliary communication were not seen in groups II and III. Group IV were 12 patients with infected cyst or intrabiliary rupture who underwent partial cystectomy with external drainage. In group IV, hospital stay was longer than in the other groups (P < 0.05). Group I had the shortest hospital stay (P < 0.05). Group IV had the highest morbidity and recurrence rates (P < 0.05). We concluded that cystectomy is the technique of choice in selected patients, as it is associated with low morbidity, low recurrence rates, and short hospital stay. Omentoplasty is preferred if cystectomy is not feasible. If there is biliary contamination and infection, external drainage, rather than omentoplasty, should be performed. PMID- 9745086 TI - Hepatectomy with microwave tissue coagulation for hepatocellular carcinoma. AB - From 1984 through 1994, 99 consecutive patients with hepatocellular carcinoma (HCC) underwent hepa-tectomy with microwave tissue coagulation (MTC). We performed limited resection (Hr0) in 28 patients, subsegmentectomy (HrS) in 25 patients, segmentectomy (Hr1) in 21 patients, and lobectomy or extended lobectomy (Hr2) in 25 patients. The patients were divided into two groups: group A, 86 patients with tumors smaller than 1 kg and no tumor thrombi in the main portal trunk; and group B, 13 patients with a tumor 1 kg or larger, or with macroscopic tumor thrombi in the main portal trunk. In group A, mean blood loss was 838 ml for Hr0, 1948 ml for HrS, 1765 ml for Hr1, and 1325 ml for Hr2. The mean operative time in group A ranged from 3 h 43 min for Hr0 to 4 h 57 min for Hr2. In group B, the mean operative time was 6 h 3 min and mean blood loss was 6053 ml. Our MTC method was associated with an in-hospital mortality rate of 3% and a major complication rate of 13.1%. The 5-year survival and disease-free survival rates were 43.4% and 25.4%, respectively. The 5-year survival rate of patients without portal tumor thrombi (50.9%) was significantly better than that of patients with portal tumor thrombi (11.9%) (P < 0.001). The 5-year survival rate of patients who underwent curative resection (58.1%) was significantly better than that of patients who underwent noncurative resection (22.9%) (P < 0.001). The 5-year survival rates of patients in group A without portal tumor thrombi did not differ between those who had cancer-negative margins (54.0%) and those with cancerpositive margins (49.6%) at resection. Recurrence and local recurrence rates did not differ in patients with cancer-positive margins (63.6% and 7.3%, respectively) and patients with cancer-negative margins (56.5% and 8.7%, respectively). These results suggested that microscopic residual cancer in the resected margin was coagulated by MTC. Blood loss, operative time, and clinical outcome in this series of 99 consecutive hepatectomies were comparable with values in earlier reports in which such hemostatic methods as the Pringle maneuver were used. We conclude that hepatectomy with MTC is useful and safe and produces consistent results. PMID- 9745087 TI - Symptomatic liver cyst: special reference to surgical management. AB - We conducted a retrospective study of 14 patients with symptomatic liver cysts to evaluate current therapeutic interventions for this condition. Abdominal pain (n = 7) or abdominal mass (n = 5) were the most frequent presentations. Three patients also had renal cyst. Percutaneous aspiration with ethanol sclerotheraphy was carried out in 4 patients and all cysts so treated diminished in size, with relief of the symptoms. One patient was treated by aspiration only and re retension occurred. Cystectomy was performed in 2 patients, unroofing in 5, and fenestration in 2 patients. All patients gained relief of symptoms, with no recurrence of symptoms. Computed tomography revealed that the cysts were diminished or were no longer observable after all the treatments. Our experience indicates that unroofing, fenestration, and cystectomy are safe and suitable procedures for treatment of the condition. Ethanol sclerotherapy may be a feasible alternative to surgical intervention in selected patients. PMID- 9745088 TI - Expression of CD44 in rat liver allografts during rejection. AB - As CD44 is believed to be a homing receptor involved in lymphoid trafficking and inflammatory responses, it is expected to be closely linked to transplant rejection. In this study, the expression of CD44 during liver transplant rejection was compared with the expression of lymphocyte-function associated antigen-1 (LFA-1) and intercellular adhesion molecule-1 (ICAM-1), which play an essential role in cell interactions and the initiation of immune responses. Male Brown Norway (BN) and Lewis (LEW) rats were used as donors and recipients, respectively. Orthotopic liver transplantation (OLTX) was done using the cuff technique of Kamada and Calne. Animals were killed on days 3, 5, and 7 after OLTX, and a piece of tissue from each of the liver grafts was obtained. Immunohistochemical staining was used to investigate the expression of CD44, ICAM 1, and LFA-1. CD44 was strongly expressed in portal areas of the rejected liver, and LFA-1 and ICAM-1 were expressed mainly on sinusoids and hepatocytes. These findings indicate that CD44 is closely involved in lymphocyte infiltration, which is dominant in portal areas, and that lymphocyte infiltration during the rejection process may involve a homing mechanism. PMID- 9745089 TI - Appraisal of surgical resection of gallbladder carcinoma with special reference to hepatic resection. AB - Carcinoma of the gallbladder a gastrointestinal malignancy with an extraordinarily poor prognosis. However, aggressive surgery, with special reference to hepatic resection, may improve survival. To prove this, we performed a retrospective analysis over an 18-year period to investigate the experience of a center that began employing liver resection in patients with gallbladder cancer in 1978. The analysis was based on patients' documentation and regular follow-up to January 1996. The standard procedures were extended cholecystectomy (cholecystectomy with lymphadenectomy and wedge hepatic resection), anatomic segmentectomy of segments IVa and V, and extended hepatectomy. Significance was assessed by the log-rank test. Thirty-nine patients were resected, curatively in 41% (n = 22; group I) and palliatively in 31% (n = 17; group 2). In 28% (n = 15; group 3) a palliative or no operation was performed. Only curatively resected patients were analyzed and followed up to January 1996. No patients in group 1 died postoperatively. The actuarial 5-year survival rate of the patients with curative resection was 55%. Four patients had stage I, two had stage II, four had stage III, and two had stage IV disease according to TNM-classification. Six of the 16 patients without lymph node metastasis survived more than 5 years. A significant difference in long-term survival was recognised between stage II and stage IV patients and between stage (pT1a)- and (look table 1b) (pT1b)-patients (P < 0.01). Diagnostic efforts should focus on detecting early stages I and II gallbladder cancer. In advanced cases, aggressive surgery, particularly with hepatic resection, is the method of choice and is successful even in patients 70 years and older. PMID- 9745090 TI - Right hepatic lobectomy and subsegmental resection of the left caudate lobe for gallbladder carcinoma involving the hepatic hilus: preservation of the ventral portion of the left caudate lobe. AB - A case of gallbladder carcinoma in a 75-year-old woman with familial hyperbilirubinemia and preoperative hepatic dysfunction is presented. Tube cholangiography through a percutaneous transhepatic biliary drainage (PTBD) catheter demonstrated a stricture and the hepatic confluence without filling of the gallbladder and showed two bile duct branches arising from the left caudate lobe. Cholangiography also disclosed that the left dorsal branch, which joined the right hepatic bile duct, was involved with tumor, while the left ventral branch, which joined the left hepatic duct, was not. Extended right hepatic lobectomy with resection of the dorsal portion of the left caudate lobe, preserving the ventral portion of the left caudate lobe, was performed. Postoperative cholangiography showed that the ventral branch of the left caudate lobe bile duct was preserved. Precise preoperative anatomic diagnosis of the biliary system in patients with hepatobiliary cancer allows successful subsegmental resection of the caudate lobe. PMID- 9745091 TI - Solitary fibrous tumor of the liver with CD 34 positivity and hypoglycemia. AB - We report a new case of solitary fibrous tumor (SFT) of the liver, an extremely rare neoplasm. Including the present case no more than ten cases are reported in the English-language literature. To date there is no definite proof of the origin of this tumor. Both mesothelial and fibroblas-tic genesis has been postulated. The monoclonal antibody CD 34 has recently been used for the characterization of SFT. SFT would appear to be histogenetically related to a CD 34 - positive fibroblastic stem cell. A 61-year-old woman was admitted to our department with epigastric and right hypochondriac pain, weight loss, and hypoglycemia. Ultrasonography and computed tomography demonstrated a large heterogeneous mass in the right hepatic lobe. A right hepatectomy was performed. The tumor weighed 2850 g and microscopic section revealed a peculiar random pattern, the so-called patternless pattern of spindle tumor cells separated by abundant thick collagen bands. The tumor presented a number of highly cellular areas composed of plump spindle cell with hyperchromatic nuclei and rare mitotic figures. Ninety percent of the neoplastic cells displayed strong immunoreactivity for CD 34/My 10. The postoperative course was uneventful and the patient is alive and well without recurrence 6 years after surgery. PMID- 9745092 TI - Hepatocellular carcinoma with chondrosarcomatous variation: case report with immunohistochemical findings, and review of the literature. AB - Hepatocellular carcinoma with chondrosarcomatous variation is very rare. We report a case with the results of pathology examination, and review the literature. The patient, a 72-year-old may had a very large tumor in the liver revealed during follow-up for diabetes mellitus. The liver mass, which was 14 cm in diameter, was diagnosed as hepatocellular carcinoma by abdominal ultrasonography. Anterior segmentectomy and partial liver resection were performed. Histopathology examination revealed that the tumor consisted of two different components: the major one was hepatocellular carcinoma (HCC), which occupied most of the tumor; and a sarcomatous component, which occupied a smaller area, and included spindle-shaped cells with chondroscarcomatous variation. Intrahepatic metastases and tumor thrombi of HCC were also found in portal and hepatic veins. Investigations of the immunohistochemical localization of keratin (KRT), vimentin (VMT), and S-100 protein (S 100) were performed by the avidin biotin complex method. Some of the spindle cells were immunohistochemically positive for both KRT and VMT, and the chondrosarcomatous cells were positive for S 100. These results strongly suggested that the sarcomatous lesion resulted from a sarcomatous change of HCC. PMID- 9745093 TI - Mucosal bile duct carcinoma with superficial spread. AB - We describe a case of mucosal bile duct carcinoma with superficial spread in a 69 year-old man with gallstone pancreatitis. The patient was seen at the hospital because of abdominal pain, fever, and jaundice. Endoscopic retrograde cholangiography (ERC) demonstrated a protruding lesion in the lower third of the common bile duct (CBD) showing wall irregularity suggestive of malignancy. Percutaneous transhepatic cholangioscopy (PTCS) disclosed a papillary tumor with granular mucosa extending continuously to the middle third of the CBD. Cholangioscopic biopsy specimens taken from both the papillary tumor and surrounding granular mucosa revealed papillary adenocarcinoma. After this assessment of extent of cancer by PTCS, we performed pancreatoduodenectomy with extrahepatic bile duct resection and regional lymph node dissection. Pathology examination revealed papillary adenocarcinoma limited to the mucosal layer. The resected margin of the bile duct was free of tumor. We also reviewed 25 cases of early mucosal bile duct carcinoma described in detail in the Japanese literature, and we discuss the diagnostic advantages of PTCS. PMID- 9745094 TI - Elevated Amniotic Fluid Interleukin-6 as a Predictor of Neonatal Periventricular Leukomalacia and Intraventricular Hemorrhage. AB - > Objective: To investigate the relationship between amniotic fluid interleukin-6 levels and the development of periventricular leukomalacia and intraventricular hemorrhage in the preterm neonate and to compare the value of amniotic fluid interleukin-6 with amniotic fluid culture and histologic chorioamnionitis in the prediction of periventricular leukomalacia and intraventricular hemorrhage. Methods: 119 women, between 20 and 34 weeks gestation, in preterm labor with intact membranes, underwent transabdominal amniocentesis. Amniotic fluid was cultured for aerobic and anaerobic bacteria, Ureaplasma urealyticum and Mycoplasma hominis. Amniotic fluid interleukin-6 levels were determined by enzyme linked immunosorbent assay. The placentas were examined for histopathologic evidence of inflammation. Where the birth weight was <2,000 g, transfontanelle cranial sonography was performed on the 3rd and 7th days of life for diagnosis of periventricular leukomalacia and intraventricular hemorrhage. Student's t test, the Mann-Whitney U test, likelihood ratio chi2, logistic regression, and receiver operator characteristic curve were used for analysis. Results: 33 women were excluded from the analysis because they delivered at other institutions. The neonates of 33 women did not have sonography because they weighed >2,000 g at birth. Two neonates died before sonography was performed; four neonates who weighed <2,000 g at birth did not have sonography. In the definitive study group of 47 women, those with neonates who developed periventricular leukomalacia and intraventricular hemorrhage (n = 14) had higher median amniotic fluid interleukin 6 levels (42,795 pg/ml versus 8,020 pg/ml; P = 0.009), more positive amniotic fluid cultures (64% vesus 21%; P < 0.003), and a shorter median amniocentesis-to delivery interval (16 h versus 24 h; P = 0.045) than women (n = 33) who delivered neonates without periventricular leukomalacia or intraventricular hemorrhage. The groups did not differ in gestational age at admission (P = 0.15), birth weight (P = 0.09), or histologic chorioamnionitis (P = 0.37). An amniotic fluid interleukin 6 level >/=20,000 pg/ml had a sensitivity of 71% and a specificity of 70% compared with a sensitivity of 69% and specificity of 79% for amniotic fluid culture, and a sensitivity of 71% and specificity of 42% for histologic chorioamnionitis in the prediction of periventricular leukomalacia and intraventricular hemorrhage. Women with amniotic fluid interleukin-6 levels >/=20,000 pg/ml (n = 20) had more neonates with periventricular leukomalacia or intraventricular hemorrhage than women with amniotic fluid interleukin-6 levels <20,000 pg/ml (n = 27) (50% versus 15%; P = 0.009). They also were of lower birth weight (P = 0.02), had more neonatal morbidity (P = 0.01), had more positive amniotic fluid cultures (P = 0.01), and more histologic chorioamnionitis (P = 0.02). Logistic regression analysis demonstrated that amniotic fluid interleukin 6 was an independent risk factor for the development of periventricular leukomalacia and intraventricular hemorrhage (odds ratio, 5.81; 95% confidence interval, 1.02-33.16; P = 0.05) after controlling for gestational age, birth weight, histologic chorioamnionitis, and amniotic fluid culture (odds ratio, 7.94; 95% confidence interval 1.22-51.77; P = 0.03). Conclusions: In women in preterm labor with intact membranes amniotic fluid interleukin-6 is useful in predicting neonatal periventricular leukomalacia and intraventricular hemorrhage. PMID- 9745095 TI - Optical Brain Monitoring of the Cerebrovascular Effects Induced by Acute Cocaine Exposure in Neonatal Pigs. AB - > Objective: The purpose of this study was to assess the direct effects of cocaine on cerebral blood flow and oxygenation in newborn piglets using optical spectroscopy. Optical spectroscopy is capable of monitoring changes in cerebral oxyhemoglobin, deoxyhemoglobin, and total hemoglobin continuously, noninvasively, and in real time. Methods: Five anesthetized and ventilated newborn piglets were injected intravenously with 1 mg/kg cocaine through a central venous catheter over 60 seconds. Cerebral blood flow and oxygenation were assessed by optical spectroscopy and standard physiologic monitoring: mean arterial pressure, carotid blood flow, cerebrovascular resistance, and arterial hemoglobin oxygen saturation (by pulse oximetry). Results: Cocaine induced a profound increase in cerebrovascular resistance, a decrease in carotid blood flow, and a decrease in mean arterial pressure and heart rate. Cerebrovascular changes were detected readily by optical spectroscopy (i.e. a decrease in cerebral oxyhemoglobin was associated with an increase in deoxyhemoglobin). The hypotensive response augmented the cerebral vasoconstriction effect on carotid blood flow. A good correlation existed between the changes in carotid Doppler blood flow and total hemoglobin in each animal and for all animals combined. Peripheral arterial hemoglobin oxygen saturation measured by pulse oximetry remained normal throughout the experiment. Conclusions: The cause for the apparent idiosyncratic hypotensive response to intravenous injections of cocaine is uncertain and requires further investigation. Our study clearly demonstrates that optical spectroscopy might become an extremely useful tool for monitoring cerebral hemodynamics and oxygenation in cocaine-exposed fetuses and infants. PMID- 9745096 TI - Maternal Brachial Arterial Waveforms in Gestational Hypertension: Analysis Using a Laplace Transform Technique. AB - > Objective: A pilot study was designed to investigate whether maternal brachial arterial Doppler shift waveforms analyzed using a Laplace transform analysis technique could distinguish women with gestational hypertension and preeclampsia from women who remained normotensive during their pregnancy. Methods: We examined 50 pregnant women in total. They were divided into two groups. The first was a group of 21 normotensive women, and the second was a group of 29 women with gestational hypertension or preeclampsia, not on medication at the time of study. A Doppler shift waveform of the right and left brachial artery was obtained and analyzed using the conventional parameters of resistance index (RI) and pulsatility index (PI) and Laplace transform analysis. Results: The systolic and diastolic blood pressures and the mean arterial blood pressure at presentation were significantly higher in group 2 as expected. The results of the Laplace transform parameters showed significant differences in the resonant frequency, real pole, c coefficient, and alpha between the two groups. There was no difference in the PI between the two groups, but the differences in RI just reached significance. The Laplace transform parameters in the group with gestational hypertension were consistent with peripheral vasoconstriction, which has been suggested as part of the hemodynamic abnormalities in this condition. Conclusion: Analysis of maternal brachial arterial Doppler shift waveforms by Laplace transform analysis detects differences between normotensive and hypertensive patients. These differences may also reflect some of the hemodynamic changes occurring in gestational hypertension. PMID- 9745097 TI - Clinical Usefulness of Flow Cytometry in Detection and Quantification of Fetomaternal Hemorrhage. AB - > Objective: To compare the technique of immunofluorescence flow cytometry against traditional Kleihauer testing with respect to the sensitivity and specificity of clinical detection of fetomaternal hemorrhage. Methods: Blood samples from D-negative unsensitized postpartum women were analyzed by flow cytometry (customized, directly conjugated monoclonal anti-D) and Kleihauer testing using commercial kits as well as a manual technique. Results: Both flow cytometry and manual Kleihauer tests performed well in vitro against known standards, giving similar results with detectable fetal cells in the range of 2 10%. Correlation of in vivo results obtained by flow cytometry and either Kleihauer method was poor. The percentage of fetal red cells detected by flow cytometry was approximately twice that of the manual Kleihauer. Commercial Kleihauer testing proved unreliable in the in vivo setting. Conclusions: Flow cytometry offers a reliable alternative to traditional Kleihauer testing for detecting fetomaternal hemorrhage and may, with standardized methodology, help optimize immunoprophylaxis against erythroblastosis fetalis. PMID- 9745098 TI - The Effect of Oral Ketanserin on Fetal Heart Rate Parameters. AB - > Objective: To determine the effects of ketanserin given from early in the midtrimester of pregnancy on fetal heart rate parameters later in pregnancy. Methods: Patients with diastolic blood pressure persistently higher than 80 mm Hg between 12 and 20 weeks' gestational age were randomized to receive either ketanserin or placebo. Both groups also got 75 mg of aspirin per day. The fetal heart rate was monitored once every fortnight from 28 weeks' gestation and weekly from 36 weeks onward. Recordings were made with the Sonicaid System 8000 and continued until the Dawes and Redman criteria were met. Results: The Dawes and Redman criteria were met within 10 min in 54% of recordings in both study groups, whereas they were not met in 4.5 and 3.3% of recordings in the ketanserin group and placebo group, respectively (P = 0.58). The mean time to meet these criteria duration per recording was 16.9 min in the ketanserin group and 16.2 min in the placebo group (P = 0.83). There were no significant differences between the two study groups in fetal heart rate variability, accelerations, or decelerations before 38 weeks' gestation. Thereafter the mean minute range and short-term and baseline variability were significantly higher in the ketanserin group, and significantly more accelerations of the fetal heart were recorded in the ketanserin group. Conclusions. Ketanserin does not influence the fetal heart rate adversely and may even be associated with improved recordings late in pregnancy. PMID- 9745099 TI - Differences between Twin Umbilical Coiling Indices Correlate to Differences in Twin Weight and Doppler Indices. AB - > Objective: The aim of our study was to evaluate sonographic and Doppler detectable differences in umbilical coiling index and fetoplacental circulation of discordant twins. Study Design: Doppler blood flow studies in 13 pairs of concordant and 20 pairs of discordant twins were performed from umbilical artery, middle cerebral artery, inferior vena cava, and ductus venosus. Flow studies were compared and correlated with the antenatal sonographic coiling index and the actual umbilical cord length, number of vascular helices, and birth weight. All studies were performed within 72 h before delivery. Pulsatility index (PI) values were calculated for the arteries and preload index (PLI) values for the veins. The umbilical coiling index (CI) was calculated using sonographic longitudinal views of cord vessels from several segments antenatally and by dividing the total number of helices by cord length (cm) postnatally. Discordancy was defined as a more than 20% intrapair actual birth weight difference. For all these index values the intertwin differences (Delta values) were calculated by subtracting the values obtained in the larger twin with those of the smaller twin. Results: The mean +/- SD intertwin difference in umbilical coiling index was 27.4 +/- 10.5% in the antepartum period and 28.9 +/- 10.0% after birth. Regression analysis showed a significant linear trend (r = 0.77, P < 0.001) between intertwin birth weight difference (DeltaBW) and intertwin coiling index difference (DeltaCI). A good correlation was found between DeltaCI and DeltaPLI in the ductus venosus (r = 0.63, P < 0.05), DeltaPLI in the inferior vena cava (r = 0.51, P < 0.005), and DeltaPI in the middle cerebral artery (r = 0.44, P < 0.05). Conclusions: Intertwin difference in antepartum umbilical coiling index can be determined by ultrasound and correlates well with: 1) the actual difference in coiling indices at birth, 2) the intertwin birth weight difference and 3) the intertwin Doppler flow characteristics in the fetal cerebral and venous circulation. PMID- 9745100 TI - Comparison between Intravenous Prostaglandin E2 and Extraamniotic Prostaglandin F2alpha Instillation for Termination in Second-Trimester Pregnancy. AB - > Objective: To evaluate the efficacy and safety of two different methods, intravenous prostaglandin E2 and extraamniotic prostaglandin F2alpha instillation, in second-trimester pregnancy termination. Methods: We designed a prospective randomized longitudinal study. 130 consecutive patients with various indications for second-trimester pregnancy termination were recruited. Patients were managed randomly with either intravenous continuous prostaglandin E2 infusion or extraamniotic prostaglandin F2alpha instillation. Laminaria were inserted in patients with unfavorable cervixes. The instillation-abortion time, success rate within 24 hours, dosage of both medications, and side effects were recorded and analyzed. Results: There was a significantly shorter instillation abortion time (11.85 +/- 9.65 versus 22.18 +/- 16.83 hours, P < 0.001), higher complete abortion rate (71.91% versus 41.5%, P = 0.013), and a higher rate of successful abortion within 24 hours (87.6% versus 56.1%, P < 0.001) in patients treated with intravenous prostaglandin E2 than in those with extraamniotic prostaglandin F2alpha. Conclusions: Prostaglandin E2 had a higher rate of successful abortion and fewer side effects than prostaglandin F2alpha. This implies that intravenous prostaglandin E2 might be a better choice for second trimester pregnancy termination compared with extraamniotic prostaglandin F2alpha. PMID- 9745101 TI - Comparisons of Nitric Oxide Synthases in Normal Human Placenta from 37 to 41 Weeks Gestation: Qualitative and Quantitative Analyses. AB - > Objective: The purpose of this study was to determine the type of nitric oxide synthase isoform and to measure the quantities of nitric oxide synthase mRNA in the human placenta. Methods: The isoforms of nitric oxide synthase were determined in ten placentas of normal pregnant women using information regarding calcium dependence and inhibition by arginine analogs and findings from Western blot analyses and reverse transcriptase-polymerase chain reaction. Results: The activity of nitric oxide synthase was largely calcium dependent, although a small element of calcium-independent activity was observed. A stronger inhibition was shown with Nomega-nitro-l-arginine than with Nomega-monomethyl-l-arginine. On Western blots endothelial nitric oxide synthase was detected as a band of 140 kilodaltons, without evidence of inducible nitric oxide synthase. Messenger RNA of endothelial nitric oxide synthase was readily detected by reverse transcriptase-polymerase chain reaction, but that of inducible nitric oxide synthase was barely detected. The quantity of mRNA of inducible nitric oxide synthase was about 100-fold less than that of endothelial nitric oxide synthase. Conclusions: These results point to the constitutive endothelial type as the predominant isoform of nitric oxide synthase in human placenta from 37 to 41 weeks gestation, although protein and mRNA of inducible nitric oxide synthase may exist. PMID- 9745102 TI - Respiratory Difficulty Necessitated Early Delivery in a Woman with Spondyloepiphyseal Dysplasia. AB - > Spondyloepiphyseal dysplasia is a skeletal disorder in which the vertebrae are flattened, thus making the spine shorter and greatly deformed, with marked thoracic kyphoscoliosis, lumbar lordosis, and a shortened trunk. The pelvis is also severely deformed and contracted. The patient was a 40-year-old nulligravida. At the 33rd week of pregnancy, respiratory distress gradually worsened, and cesarean section under spinal anesthesia had to be done. In women with spondyloepiphyseal dysplasia, uterine expansion is limited. As respiratory difficulties increase, an early delivery may have to be done. PMID- 9745103 TI - Outcome of Two Pregnancies in a Patient with Gitelman's Syndrome-A Case Report. AB - > Gitelman's syndrome is primary renal tubular hypokalemic metabolic alkalosis with hypocalciuria and magnesium deficiency. We present the prenatal course and outcome of two pregnancies in a patient with Gitelman's syndrome. The complication encountered was oligohydramnios. Close monitoring of serum magnesium and potassium, amniotic fluid volume, and fetal growth is required. PMID- 9745104 TI - Glycosylphosphatidylinositol-anchored proteins in human and pig's milk. AB - Glycosylphosphatidylinositols (GPIs) are a recently discovered class of glycoconjugates that anchor either proteins, polysaccharides or small oligosaccharides to cellular membranes via a covalent linkage. To investigate the presence of soluble GPIs, individual human milk samples and mature pig's milk were defatted and casein removed by acid precipitation and ultracentrifugation. Soluble proteins were subjected to FPLC gel filtration (Superdex 200) and high molecular weight proteins were separated into fractions I-V. Immunological studies have been performed using Western blotting of whole milk, casein and whey fractions, and Superdex fractions I-V followed by incubation with a monoclonal antibody against the purified GPI anchor of the variant surface glycoprotein from Trypanosoma brucei brucei. We found that significant amounts of GPI-anchored proteins are secreted into human milk and pig's milk. The antibody which reacted only with components in the whey fractions bound to 5 different human milk proteins with a molecular weight of >/=200 (at least 3 components), 90 and 80 kD. In pig's milk, the staining pattern was found to be different from human milk. Similar to other GPI-anchored cell structures the functions of GPI-containing proteins in human milk and pig's milk as well as the specific components carrying these anchors remain to be investigated. PMID- 9745105 TI - Effects of dietary long-chain polyunsaturated fatty acids on plasma amino acids and indices of protein metabolism in infants: results from a randomized clinical trial. AB - BACKGROUND/AIM: Previous studies in vitro and in animals in vivo found that alpha linolenic acid (C18:3omega3) may enhance oxidative damage of essential amino acids. We investigated whether the addition of the long-chain polyunsaturated fatty acids (LCPUFA) arachidonate (C20:4omega-6; AA) and docosahexaenoate (C22:6 omega3; DHA) in the form of egg phospholipids to infant formula affects plasma amino acid concentrations and indices of protein metabolism in term infants. METHODS: In a double-blind, randomized clinical trial, healthy infants were fed from day 5 of life formula with or without preformed LCPUFA (n = 10 and 12, respectively). At the age of 5 days and 1, 2, 3 and 4 months, blood samples were obtained and analyzed for plasma amino acids by high-performance liquid chromatography and for plasma phospholipid fatty acid composition by gas chromatography. RESULTS: At the age of 3 months, plasma threonine concentrations were significantly lower in infants receiving dietary LCPUFA than in controls (124 +/- 16 vs. 216 +/- 28 micromol/l, p < 0.05). Values of other plasma essential amino acids, total protein, albumin, creatinine and urea nitrogen did not differ between the two feeding groups throughout the study. At the age of 5 days, plasma phospholipid AA and DHA concentrations were inversely correlated with histidine concentrations (AA: r = -0.60, p = 0.01; DHA: r = -0.53, p < 0.05). At the age of 3 months, DHA concentrations were inversely related to plasma histidine, methionine and threonine concentrations (r = -0.66, -0.62, and 0.64, respectively, p < 0.05). CONCLUSIONS: The dietary LCPUFA supplementation of infant formula used in this study has no adverse effects on infant plasma amino acid concentrations and indicators of protein metabolism. Nonetheless, the apparent interaction of LCPUFA with some amino acids in formula-fed infants warrants further investigation. PMID- 9745106 TI - Apolipoproteins A-I and B in Kuwaiti children. AB - To assess the relation of apolipoproteins (Apos) A-I and B (the carrier proteins for high and low density lipoprotein cholesterol, respectively) with the degree of obesity, body fat distribution, serum lipids, glucose and insulin levels, a case-control study was carried out and included 460 Kuwaiti obese children, 6-13 years old, matched by age and sex to 460 normal-weight controls. Obese children were ascertained in a representative cross-sectional study of 2,400 school children. The Apo A-I levels were not different between obese and non-obese boys, while they were significantly lower in obese girls (p < 0.01). The Apo B mean concentrations were significantly higher in obese boys and girls (p < 0.001), while the Apo A-I:B ratio was significantly lower in obese children (p < 0.001). Apo A-I levels were positively correlated with total cholesterol, high- and low density lipoprotein cholesterol, but were not correlated with very low-density lipoprotein cholesterol, triglycerides, insulin, glucose or insulin:glucose ratio. Apo B levels were negatively correlated with high-density lipoprotein cholesterol and positively correlated with insulin and insulin:glucose ratio (p < 0.01) in obese children. The study documented an adverse Apo profile in obese Kuwaiti children. Since Apo changes are correctable through management of obesity, their identification in childhood offers prospects for prevention of early onset atherogenesis in adulthood. PMID- 9745107 TI - Effects of consumption of oat milk, soya milk, or cow's milk on plasma lipids and antioxidative capacity in healthy subjects. AB - A drink based on oats has been developed with new technology. In this study the effects of this oat milk, soya milk and cow's milk on plasma lipid, glucose, insulin, and antioxidant status (measured as the ability of serum to suppress the formation of the radical cation ABTS*+) were compared in 24 healthy men and women. Half of the subjects (group A) consumed 0.75-1 liters/day of oat milk and soya milk for 4 weeks each, and the other half (group B) consumed oat milk and cow's milk for two 4-week periods. In the combined groups A plus B the oat milk regimen resulted in decreased plasma cholesterol (4%) and low-density lipoprotein (LDL) cholesterol (9%) levels as compared with baseline, but no changes in high density lipoprotein cholesterol (HDL) and triglyceride values were observed. Also soya milk consumption resulted in decreased LDL cholesterol concentrations. The only significant plasma lipid change observed during consumption of cow's milk was an increase in HDL cholesterol. No consistent changes in body weight, fasting blood glucose, serum insulin, and antioxidant status occurred after consumption of any milk regimen. A significant correlation between baseline antioxidant status and total plasma cholesterol was found (r = -0.56). It is proposed that the high content of beta-glucans in oat milk was responsible for the decreased plasma cholesterol and LDL cholesterol concentrations, but the effect could also be due to a replacement of saturated fat in the customary diet by unsaturated fat. It is concluded that oat milk can be used as an alternative to other milk drinks by subjects who would benefit from reduced LDL cholesterol values. PMID- 9745109 TI - Analysis of fecal bile acids and neutral steroids using gas-liquid chromatography. AB - In the present pilot study, for investigating the physiological effects of different types of nondigestible oligosaccharides, we have validated the application of methodologies for the analysis of bile acids and neutral steroids in feces of human subjects. The accuracy of the extraction and chromatographic procedures for the analyses of bile acids and neutral steroids was determined by recovery of added compounds to fecal homogenate. The precision of the above procedures was checked by analyzing these compounds in samples (n = 5) of the same fecal homogenate. Recoveries of added bile acids ranged from 86 and 96%, and those of neutral steroids varied from 81 to 97%. The precision expressed as coefficients of variation of bile acids and neutral steroids ranged from 2.3 to 8.3% and from 6.3 to 11.8%, respectively. The intra- and interindividual variabilities expressed as coefficients of variation of bile acids varied from 1 to 58 and from 0 to 74%, respectively. The same variabilities for neutral steroids ranged from 0.5 to 107% and from 1 to 168%, respectively. The methods validated in the present pilot study were adequate for applying to our forthcoming European Union coordinated major study on the physiological effects of different types of nondigestible oligosaccharides and involving large numbers of samples. PMID- 9745108 TI - Absorption of cholesterol oxidation products from ordinary foodstuff in humans. AB - BACKGROUND: Information on the absorption of cholesterol oxidation products (COP) from ordinary foodstuff in humans is scarce. METHODS: Five healthy young men were offered a salami- and Parmesan-containing meal naturally rich in COP. Plasma and lipoprotein COP concentrations were measured over the following 9 h. RESULTS: The mean plasma free (nonesterified) COP concentration showed its maximal increase 3 and 5 h after meal consumption. In contrast, the raise in plasma total COP concentration began 6 h after the meal with a maximum at 8 h and was statistically significant for 7alpha- and 7beta-hydroxycholesterol and 7 ketocholesterol. The increase in plasma total cholesterol concentration was comparable to that of total COP. Comparing the COP composition of the chylomicrons and the test meal, cholestanetriol, 7-ketocholesterol, and to a lesser extent cholesterol-alpha-epoxide were underrepresented in the chylomicrons as was the opposite for 7beta-hydroxycholesterol. In very-low-density lipoprotein, a steady increase in the COP:cholesterol ratio was observed from 6 h on. CONCLUSION: COP from ordinary foodstuff were absorbed in the human intestinal tract but differences in the bioavailability of the single COP compounds were found. PMID- 9745110 TI - Effects of aging on the energy balance of pregnant rats. AB - In pregnant and non-pregnant female rats at various ages the energy balance was determined to study the age-induced alterations in the mother and conceptus. The animals were anesthetized and the conceptus removed and separated into male, female, and placental tissues. The animals were then killed and the carcasses prepared for energy balance determination. The results obtained showed that the older animals gained less energy in the body and had lower gross food efficiency than the younger animals. The number, energy and weight of the offspring were not affected by the mother's age. Nevertheless, due to its constant growth during life, the rat is not the best model to study age-related processes. PMID- 9745111 TI - A linoleic acid enriched diet increases serum cholesterol esterification by lecithin:cholesterol acyltransferase in meal-fed rats. AB - Dietary fats are known to influence the fatty acid profile of plasma lipids, including phospholipids which are substrates of lecithin:cholesterol acyltransferase (LCAT; EC 2.3.1.43), an important enzyme in lipoprotein metabolism. We tested whether the dietary fatty acid profile has an effect on LCAT activity in an animal model. Rats were conditioned to eat two meals per day, which were enriched in either palmitic, oleic or linoleic acids, for 10 weeks. Serum was isolated from blood samples taken prior to the meal. The LCAT activity was determined in two ways: (1) by measuring serum cholesterol esterification rates, which are an estimate of LCAT action on endogenous lipoproteins, and (2) by measuring serum LCAT activity levels with excess exogenous substrates, an estimate of LCAT mass. Animals receiving the linoleic acid diet had lower serum concentrations of unesterified cholesterol and triglycerides, if compared with animals fed oleic acid or palmitic acid diets (p < 0.05). Serum LCAT activity levels (measured with excess exogenous substrates) were not different, but both the absolute and fractional rates of cholesterol esterification were highest on the linoleic acid rich diet (p < 0.01), showing that LCAT action on endogenous lipoproteins is improved. No differences were found in serum apolipoprotein B and A-IV concentrations between the dietary groups. Apolipoprotein A-I levels were lowest in the palmitic acid group (oleic and linoleic > palmitic; p < 0.05), and apolipoprotein E levels were highest in the palmitic acid group (palmitic > oleic and linoleic; p < 0.05). It is concluded that a linoleic acid rich diet may cause increased metabolism of serum cholesterol by LCAT in rats. This effect is not due to elevated serum concentrations of LCAT or of its apolipoprotein activators, but most likely to changes in the chemical composition of endogenous lipoprotein substrates. It remains to be established whether the serum cholesterol esterification rates measured in vitro are related to in vivo rates of reverse cholesterol transport. PMID- 9745112 TI - Developmental enamel defects in children with different fluoride supplementation- a follow-up study. AB - The intake of fluorides with water, food, dental fluoride preparations, or in particular fluoride supplements, such as NaF tablets, may lead to dental fluorosis. In the present study conducted in a nonfluoridated area in Germany, developmental enamel defects were examined using the Modified Developmental Defects of Enamel Index (Mod DDE Index), which subdivides enamel defects into the categories demarcated (Mod DDE score 1) and diffuse (Mod DDE score 2) opacities and hypoplasia (Mod DDE score 3). 158 children, between 8.5 and 10 years old, were assigned to three examination groups, defined by three different fluoride tablet programs. The children in all three examination groups, F1, F2, and F3, had received 0.25 mg F-/day up to the age of 2 years, F1 and F3 from birth on, F2 beginning with the 7th month of life. F1 and F2 received 0.5 mg F-/day during the 3rd and 0.75 mg F-/day during the 4th and 5th year of life. For F3, beginning with the 3rd year of life, no further recommendations were made. 158 sociodemographically matched children living in a neighboring town served as controls and did not take part in any structured fluoride supplementation program. The proportions of children with Mod DDE scores 1, 2, or 3 at least in one index tooth were significantly higher in the examination groups (40%) than in the control group (20%). Also, the proportions of children with Mod DDE score 2 at least in one index tooth were significantly higher in the examination groups (18%) than in the control group (8%). The proportions of children with Mod DDE score 1 at least in one index tooth were 25% in the examination groups and 17% in the control group. No Mod DDE score 3 was found. Not more than 5% of the children in each group had 50% of their teeth with Mod DDE score 1, 2, or 3. The proportions of teeth per child with Mod DDE score 2 were significantly higher in the examination group than in the control group. While uncontrolled variables cannot be excluded, the observed differences between the experimental and control groups may be attributed to the ingestion of fluoride tablets in the experimental group. PMID- 9745113 TI - Fluoride concentration in whole saliva and separate gland secretions in schoolchildren after intake of fluoridated milk. AB - Fluoride concentration in whole saliva and in separate gland secretions was studied after a 7-day fluoridated milk regimen (1mg F per day) in 12 healthy schoolchildren aged 10-13 years. A 2-week fluoride-free run-in period preceded the tests in order to establish the endogenous baseline levels. Unstimulated and stimulated whole saliva and stimulated parotid and submandibular-sublingual saliva were collected at 1, 3, 6, 12 and 24h after F-milk ingestion, and fluoride concentrations were determined with an ion-selective electrode. Typical time dependent excretion curves were obtained in all collected secretions. The fluoride levels were significantly elevated 1 and 3h in whole saliva and up to 6h in the gland secretions after intake of fluoridated milk when compared to baseline values. When acid-stimulated, the submandibular-sublingual glands were the major contributors of fluoride in the oral cavity. In conclusion, the results of this study demonstrate that fluoride ingested with milk is excreted through the salivary glands, indicating that the bioavailability of fluoride from milk equals that of other vehicles. PMID- 9745114 TI - Fluoride uptake to demineralised enamel: A comparison of sampling techniques. AB - Fluoride uptake is a recognised way of assessing the potential anticaries efficacy of fluoride (F) treatments. The aim of the present study was to compare an abrasion method of sampling treated enamel, based on that of Weatherell et al. [Caries Res 1985;19:97-102], with the acid-etch method of Raven et al. [Caries Res 1991;25:130-137]. Two adjacent demineralised areas were created on the polished surfaces of bovine incisors using an acid gel system. One artificial lesion from each tooth was subsequently treated for 6 h at 37 degrees C with one of two fluoridated dentifrice slurries (1 part: 3 parts water), whereas the other was treated similarly with a slurry of non-F control dentifrice. One set of treated lesions was then separated, the base of each enamel block polished until planoparallel with the demineralised surface and the lesions isolated by cutting away the adjacent sound enamel. Each block was mounted on the probe of a digital micrometer and the demineralised surface abraded with silicon carbide lapping film until sound material was reached. Abraded material was dissolved in perchloric acid and the buffered solution analysed for fluoride by ion-selective electrode. Each lesion of a second set of treated, demineralised enamel blocks was etched by 20microl acid and the resulting solutions analysed for F. Mean F uptakes [microg cm-2 (SD)] were: abrasion (n = 7/treatment); F dentifrice A = 1.39 (0.89) and B = 0.86 (0.45) relative to non-F controls = 0.11 (0.12), 0.14 (0.06), respectively; and acid-etch (n = 14/treatment); A = 1.27 (0. 49), B = 0.69 (0.23), controls = 0.12 (0.06), 0.12 (0.06), respectively. Significant differences (p<0.05) for both data sets were: A>B> control. The results show good agreement between the sampling methods and demonstrate the ability of the abrasion technique to distinguish between F treatments. PMID- 9745115 TI - Effect of toothpicks with and without fluoride on De- and remineralization of enamel and dentine in situ. AB - The aim of this investigation was to study the effect of the interproximal use of fluoride (F)-impregnated and non-impregnated birch toothpicks on the degree of de and remineralization of enamel and dentine in situ. Ten volunteers with complete dentures in the upper jaw participated. Each subject had four specimens: (1) sound enamel, (2) demineralized enamel, (3) sound dentine and (4) demineralized dentine; placed pairwise at two approximal sites (15/16 and 25/26) of the maxillary prosthesis. The study involved three test periods (A, B and C), each lasting 4 weeks. In A, the subjects used F toothpicks (impregnated in 4% NaF) and, in B, nonimpregnated toothpicks 3 times daily. During period C, no toothpicks were used. Dentifrice or other F-containing products were not allowed during the 4-week periods. Transversal microradiography was used to determine lesion depth (ld) and mineral loss (DeltaZ). The results revealed that all the sound samples lost mineral during the three experimental periods; DeltaZ for both enamel and dentine was less for A and B compared with C (p<0.01) and less for A compared with B and C for dentine (p<0.05, p<0.01). The demineralized samples also lost mineral, apart from dentine, during periods A and B, i.e. when F impregnated and non-impregnated toothpicks were used; ld for enamel and DeltaZ for dentine were less for A compared with C (p<0.05). Four weeks' use of toothpicks, especially F-impregnated toothpicks, thus reduces the demineralization of enamel and dentine at approximal sites in situ. PMID- 9745116 TI - Cells for the study of acidic dissolution in packed apatite powders as model systems for dental caries. AB - A number of different designs of cells have been developed for the study of dissolution processes in packed apatite powders. Basic design requirements were that no deleterious processes, such as high temperature sintering or binding agents, were involved, and that there was maximum opportunity for experimental study. One design used alternating filter paper discs and apatite layers (typically 42 mg). At the end of an experiment, the cell could be disassembled and infrared spectra and X-ray powder diffraction patterns made of individual layers. Cells without filter paper discs were also made, but terminal sampling at well-defined depths was more difficult or impossible. The cells were constructed from poly(methylmethacrylate) so the course of dissolution could be monitored by radiography. Subsurface loss of apatite was almost always seen after from 2 weeks to 8 months exposure to buffer solutions at pH 3.0-5.5. The greatest loss of apatite was typically 0.4-2 mm below the surface, which is at a larger distance than usually seen for dental enamel. This may be attributable to the low packing density (typically 35-60 vol%) found in the present systems compared with enamel. PMID- 9745117 TI - Fluoride-dependent formation of mineralized layers in bovine dentin during demineralization in vitro. AB - Demineralization of dentin in the presence of fluoride produces lesions with a mineralized surface layer which becomes thicker and more mineralized with higher fluoride concentrations whereas the lesion depth is hardly affected. The aim of this study was to investigate the effects of the time of fluoride treatment and the amount of fluoride taken up on the properties of the mineralized layer. Discs of bovine dentin embedded in methylmethacrylate with one surface exposed were demineralized in 50 mM acetic acid, 2.2 mM CaCl2, 2.2 mM KH2PO4, pH 5.0. At the start and/or later during the demineralization period, the specimens were incubated individually for 1 or 2 days in 10 ml of the same demineralization solution supplemented with 0.5, 2.0 or 5.0 ppm fluoride, which was then assessed for changes in calcium and fluoride concentrations. After 2, 5 and 8 days, specimens were sectioned for microradiographic analysis so as to follow development of the lesions and the mineralized layers. The results were the following: While demineralization with fluoride present at the first day led to the formation of a surface layer, fluoride present only at a later day produced a subsurface layer, not at the lesion front but closer to the surface. This layer resulted from (re)precipitation and not from preservation of the original mineral. The 'integrated mineral content' of the surface layer increased linearly with the uptake of fluoride, which resulted in an apparent fluorapatite content of about 20 vol%. The profiles of the surface layers remained unchanged during continued demineralization in the absence of fluoride. It was concluded that in the presence of fluoride mineral loss is reduced as a result of the reprecipitation of dissolved mineral ions as a layer of fluoride-enriched apatite. This layer does not offer protection of underlying dentin against continued demineralization. PMID- 9745118 TI - Efficacy of sterilisation methods and their effect on enamel demineralisation. AB - The aim of this project was to determine the effectiveness of sterilisation methods for dental enamel for use in intra-oral cariogenicity tests, and their possible effect on the degree of demineralisation of enamel. Bovine incisors were cut vertically into five portions and each assigned to one of five groups. Group 1 was used as a control while the other four groups were subjected, respectively, to gamma irradiation ( congruent with 25kGy), steam autoclaving (121 degrees C for 15 min), sodium hypochlorite (12% w/v for 24h) and povidone-iodine (7.5% w/v for 24h). Total viable counts of microorganisms remaining following sterilisation of the specimens were performed following incubation of the specimens for 24h at 37 degrees C. Caries-like lesions were produced in each specimen using an acidic buffer solution (pH4.5). Sections were cut from each specimen, ground to 80 microgram thickness, and microradiographed. Mineral loss and lesion depth were quantified using transverse microradiography. Statistical analysis was by ANOVA. Dunnett's and Tukey's tests. Microbial growth (Staphylococcus aureus and bacilli) was observed only in control specimens in both brain heart infusion broth and on blood agar plates. The sterilisation methods affected the enamel surface as follows: gamma irradiation (cream discolouration), NaOCl (bleaching), and povidone-iodine (white spot-like lesion). Compared with the control, there was no significant difference in mineral loss and lesion depth with any of the groups, but the numerical values of mineral loss and lesion depth in groups can be ranked as follows: gamma irradiation 70 x 10(-5)%; P. aeruginosa, >/=270 x 10(-5)%, and C. freundii, >/=550 x 10(-5)%. With 2% FCS, the lowest AgNO3 concentration studied (7 x 10(-5)%) showed cytotoxic effects (cell survival 71 +/- 19%) at only 2 h of incubation. Under these conditions AgNO3 cytotoxicity was time- and concentration dependent in all exposure periods. Cytotoxicity was greatly enhanced causing 76% fibroblast growth inhibition at concentrations of 14 x 10(-5)% and contact time of 2 h. The AgNO3 concentration of 7 x 10(-5)% was also cytotoxic with 5% FCS in the media compared with controls, although cell survival was higher than with 2% FCS. The cytoprotective action of FCS was clearly shown at the concentration of 10% at which AgNO3 cytotoxicity of 7 x 10(-5)% to 28 x 10(-5)% was partially or completely inhibited. Our results show that AgNO3 at concentrations 100-700 times more diluted than that normally used in clinical practice retained effective inhibitory activity against some of the above-mentioned microorganisms. However, even these concentrations are cytotoxic for cultured fibroblasts. Thus, silver nitrate concentrations up to 100 times more diluted can be used, since they possess bacterial growth-inhibiting power, are less cytotoxic and therefore favour wound healing. PMID- 9745142 TI - The induction of terminal differentiation markers by the cAMP pathway in human HaCaT keratinocytes. AB - The terminal differentiation of human epidermal keratinocytes is a complex morphological and biochemical shift from a mitotically active cell to an inert protein cross-linked envelope. This transition is a clearly predetermined cell death mechanism, but it is unlike many other programmed cell deaths in that it is not apoptotic. To explore and contrast the mechanism by which keratinocytes are committed to differentiation rather than apoptosis, we focused on the cyclic adenosine monophosphate (cAMP) signaling pathway using selective modulators of intracellular cAMP levels. Markers of differentiation were assayed by Western blotting. Raising intracelluar cAMP levels by treating HaCaT cells with forskolin, a diterpene, or with isobutylmethylxanthine, a phosphodiesterase inhibitor, and isoproterenol, a beta-adrenergic receptor agonist that selectively activates adenylate cyclase, increased the levels of the differentiation markers keratin K1 and K10, involucrin and transglutaminase. H89 and KT5720, both inhibitors of cAMP-dependent protein kinase, suppressed the expression of keratins K1 and K10. These observations are in line with the defined role for cAMP in the control of keratinocyte differentiation. PMID- 9745143 TI - Vitamin D3 and its synthetic analogue secocholestra-trien-1,2, 24-triol influence the metabolism and the isomerization of retinoic acid in human keratinocytes. AB - Vitamin D and vitamin A acid share metabolic pathways thereby influencing their benefit as a given drug. Little is known concerning their metabolic interactions in epidermal cells. We compared the influence of 1,25-dihydroxycholecalciferol (vitamin D3 - VD3) and its synthetic analogue secocholestra-trien-1,3,24-triol (tacalcitol - TAC) in combination with different calcium concentrations (Ca) on the metabolism and the isomerization of retinoic acid (RA) in cultured primary human keratinocytes. After preincubation with 0.09, 0.6 and 1.2 mM Ca for 24 h, followed by the addition of 10(-6), 10(-8) or 10(-10) M VD3 or TAC, we added 10( 5) M 13-cis-RA (isotretinoin). 24 h later, concentrations of RA isomers and oxidated RA metabolites were measured by RP-HPLC. VD3 (10(-6) M) inhibited the isomerization of 13-cis-RA to all-trans-RA (tretinoin) and 9-cis-RA independently from the Ca concentration in the culture medium. 10(-6)-10(-10) M TAC equally inhibit the 4-hydroxylation of all-trans-RA significantly (12.8 vs. 6.7% of total RA), thereby reducing the amount of irreversible inactivated 4-oxo-all-trans-RA, leading to a higher persistence of all-trans-RA, the active hormone. Both VD3 and its analogue TAC influence the metabolism of RA, a well-known regulator of epithelial cell proliferation and differentiation processes, in two distinct ways. Further studies are necessary to test the hypothesis that the hormone activity of RA can be increased by concomitant treatment with VD3 which prolongs the persistence of 13-cis-RA, and TAC elevating the intracellular concentration of all-trans-RA. PMID- 9745145 TI - Dermatopharmacy. halle/s. (germany), department of dermatology of the martin luther-university, november 20, 1996 PMID- 9745144 TI - 'Retinoids '97': the meeting of the European retinoid research group (ERRG) 1997. PMID- 9745146 TI - Transfusion practices in primary total joint replacements in Finland. AB - BACKGROUND AND OBJECTIVES: A multicenter survey was carried out in 11 Finnish hospitals to determine the prevailing transfusion practices in orthopedic surgery. MATERIALS AND METHODS: The records of 1,161 patients who had undergone primary total hip (THR) or knee replacement (TKR) were reviewed. RESULTS: Allogeneic red cells (RBC) were administered to 92% of the THR patients and to 84% of the patients undergoing TKR. Significant interhospital variation in RBC transfusion, from 2.4 to 4.1 units per patient in THR and from 1.8 to 2.8 units in TKR, was detected after adjustment for clinical and laboratory findings. CONCLUSION: The use of RBCs in orthopedic surgery was generous compared with reports from other European countries and the US. Each hospital's individual transfusion policy seemed to be a strong determinant of allogeneic blood use. PMID- 9745147 TI - Effects of blood donation on the physical fitness and hemorheology of healthy elderly donors. AB - BACKGROUND: International regulations for blood donation recommend a maximum donor age of 65 years. As the average population age is steadily rising in western societies, a considerable group of volunteers is lost to the donor base. STUDY DESIGN AND METHODS: In a prospective study we investigated the effect of a 450-ml whole blood donation on the physical fitness and hemorheology of regular elderly allogeneic blood donors (n = 24, aged 63-69 years, mean = 65). Results were compared with a younger group of regular donors (n = 23, aged 55-62 years, mean = 58) and a group of elderly subjects (n = 7, aged 63-66 years, mean = 65), who did not donate blood for this study. Assessing the physical fitness, we determined the submaximal physical working capacity at a heart rate of 130 min-1 (PWC 130) and the maximal working capacity (MWC) by treadmill exercise testing the day before (day -1) and after donation (d +1). The impact of the blood loss on hemorheology was examined by analyzing the plasma viscosity before, during and after donation. RESULTS: We found an increase of mean values of PWC 130 and MWC on day +1 in all study groups, but increases were only significant in the younger group (PWC 130 p = 0.03; MWC p = 0.04). Values did not differ significantly between the three groups. Plasma viscosity decreased significantly directly after donation in both groups of donors. CONCLUSION: A single blood donation did not alter the physical fitness of otherwise healthy elderly people. The older blood donors and the younger controls showed a similar compensation mechanism to blood loss. We found no general reason for disqualifying blood donors aged 65 years from donating. PMID- 9745148 TI - Impact of storage at 22 degrees C and citrate anticoagulation on the cytokine secretion of mononuclear leukocytes. AB - BACKGROUND AND OBJECTIVES: Peripheral blood mononuclear cells (PBMC) and cytokines accumulating in platelet concentrates (PC) during storage have been implicated in causing non-hemolytic transfusion reactions, in particular after prolonged PC storage. We investigated the impact of major storage parameters, such as storage time, anticoagulation and temperature, on the capability of PBMC to secrete cytokines. MATERIALS AND METHODS: In a first study, PBMC from whole blood donations (n = 16) were exposed to standard PC storage conditions: 22 degrees C, citrate phosphate dextrose (CPD) anticoagulation. Secretion of the cytokines interleukin-1 beta (IL-1 beta), IL-2, IL-6 and interferon-gamma (IFN gamma) on mitogenic stimulation was measured directly after blood donation and after 1, 3 and 5 days of storage. In a second study, paired whole blood samples (n = 24) were investigated for the mitogenic induction of IL-2, IL-6, IFN-gamma and IFN-alpha. Cytokine values were compared with respect to incubation temperature (22 vs. 37 degrees C) and anticoagulant (CPD vs. lithium-heparin). RESULTS: PBMC stored at 22 degrees C with CPD anticoagulation showed an altered cytokine secretion pattern in comparison to freshly donated cells. After storage for 3 days, IL-2 release was decreased (p < 0.05) and after 5 days secretion of all cytokines was significantly decreased, though still detectable (p < 0.01). In the second study, CPD anticoagulation resulted in a significantly impaired cytokine secretion compared with lithium-heparin. Cytokine secretion at 22 degrees C were significantly lower (p < 0.001) compared with 37 degrees C for both anticoagulants. CONCLUSION: Storage of PBMC at 22 degrees C with CPD leads to an impaired cytokine response, but PBMC are still capable of secreting cytokines after 5 days of storage. PBMC remain a potential source of cytokines even after 5 days of storage in CPD at 22 degrees C. PMID- 9745149 TI - Kinetics of bradykinin levels during and after leucocyte filtration of platelet concentrates. AB - OBJECTIVE: To measure bradykinin levels in platelet concentrates during and after leucocyte filtration. METHODS: Platelet concentrates were leukocyte depleted using three different leucocyte-depletion filters selected to represent filters with negative, positive or neutral charge, respectively. Bradykinin levels were analysed before, during and up to 90 min post-filtration. RESULTS: Significant levels of bradykinin were generated when negatively charged leucocyte depletion filters were used. However, we found a high variation between platelet concentrates prepared from different donors as well as within the same concentrate when this was divided into three aliquots. Generated bradykinin was rapidly degraded in the collection bag and no bradykinin was detectable 60 min post-filtration. CONCLUSION: Bradykinin generation is more pronounced when negatively charged leucocyte removal filters are used. Due to a rapid degradation of bradykinin in the storage bag, pre-storage filtration should minimise the risk of bradykinin related adverse reactions during transfusion with some filters. PMID- 9745150 TI - Lactated Ringer's solution versus hydroxyethyl starch for volume replacement in autologous blood donors with cardiovascular disease: a controlled, randomized trial. AB - BACKGROUND AND OBJECTIVES: The study was designed to evaluate whether volume replacement following blood donation can prevent arterial hypotension in autologous blood donors with cardiovascular disease. MATERIALS AND METHODS: One hundred nineteen autologous blood donors with known cardiovascular disease were randomly allocated to receive, following withdrawal of 500 ml of blood, either no infusion (control group) or a 25 ml/min intravenous infusion of either 1,500 ml of lactated Ringer's solution (LRS) or 500 ml of 6% hydroxyethyl starch (HES). Starting before phlebotomy, arterial blood pressure was measured oscillometrically every 5 min until 90 min after donation. RESULTS: Group means showed little difference between the groups in blood pressure throughout the monitoring period. The proportion of patients who at least once had a > or = 20% decrease from baseline in systolic blood pressure was 3-5 times greater in the control group than in the LRS and the HES group (50 vs. 10 and 15%, respectively; p < 0.001 on chi 2 analysis for a 2 x 3 table). Systolic hypertensive episodes (> or = 20% increase over baseline) were observed more frequently in the LRS group than in the control and the HES group (41 vs. 10 and 18%, respectively; p = 0.003). CONCLUSION: Both LRS and HES, administered at a volume ratio to blood loss of 3:1 and 1:1, respectively, significantly reduced the incidence of systolic hypotensive episodes in autologous blood donors with cardiovascular disease. LRS at a 3:1 volume ratio to blood loss was associated with a high rate of systolic hypertension. PMID- 9745151 TI - Use of overlapping synthetic peptides to characterize samples from blood donors with indeterminate results to hepatitis C virus core antigen. AB - BACKGROUND AND OBJECTIVES: Despite recent improvements in supplemental assays, isolated reactivity to the hepatitis C virus (HCV) core antigen continues as one of the main problems in the confirmation of anti-HCV in blood donors. Reactivity against individual peptides from the c22-3 HCV recombinant antigen has been described as a useful tool for anti-HCV confirmation and donor counseling in such cases. MATERIALS AND METHODS: We used a previously described set of overlapping peptides spanning the entire sequence of the c22-3 antigen to study 87 single serum samples from blood donors with reactivity for c22-3 antigen alone in second generation recombinant immunoblot assay (RIBA-2). All of them had been previously studied by RIBA-3, anti-HCV E2 EIA and HCV PCR. RESULTS: 66 of the 87 samples studied could be classified as positive or negative for anti-HCV core using the multipeptide assay and such classification correlated well with the results obtained with RIBA-3 and the anti-E2 EIA. However, some discrepancies were found. The epitopes located along the N-terminal half of the molecule were mainly responsible for the specific antibody recognition but those enclosed within amino acids 1-15 were frequently involved in nonspecific reactivity. Some 38% of samples were considered to have specific antibody to the c22-3 antigen and a further 9% reacted for both anti-E2 and single core peptides that were often involved in specific antibody recognition. CONCLUSION: Testing of blood donor samples indeterminate to the HCV c22-3 antigen for reactivity against individual core peptides can confirm the presence of specific antibody and recognize nonspecific reactivity with certain cross-reacting epitopes. Third generation supplemental tests have reduced such false reactivity, but confirmation of samples with anticore alone is still necessary. Single reactivity to the HCV core antigen is likely to reflect prior exposure to the virus, but rarely active infection, either acute or chronic. PMID- 9745152 TI - Point mutations and deletion responsible for the Bombay H null and the Reunion H weak blood groups. AB - OBJECTIVE: Definition of the molecular basis of the Reunion and the Bombay red cell and salivary H-deficient phenotypes. METHODS: Sequence and expression of FUT1 and FUT2 genes from H-deficient individuals. Family segregation analysis of the mutations responsible for the fucosyltransferase defects of H, secretor and Lewis systems. RESULTS: The Indian red cell H null Bombay phenotype depends on a new mutation of the FUT1 gene. T725-->G changing Leu242-->Arg. Their salivary nonsecretor phenotype is secondary to a complete deletion of the FUT2 gene. The red cell H weak Reunion phenotype depends on another new mutation of FUT1, C349- >T which induces a change of His117-->Tyr. Their salivary nonsecretor phenotype is due to the known Caucasian inactivating mutation G428-->A. CONCLUSION: Single prevalent FUT1 and FUT2 point mutations and a deletion are responsible for the Indian Bombay H null and the Reunion H weak phenotypes found on Reunion island. This is in contrast with other H-deficient phenotypes where sporadic nonprevalent inactivating mutations are the rule. PMID- 9745154 TI - RHD gene polymorphisms among RhD-negative Chinese in Taiwan. AB - BACKGROUND AND OBJECTIVE: The rare occurrence of anti-D-associated hemolytic disease of the newborn among Chinese is attributable in part to the existence of the weak D phenotype Del among apparently RhD-negative individuals. While existing advances in the molecular genetics of the Rh blood group have been noted in recent years, the genomic structure of the Del phenotype has seldom been studied in the literature. We try to explore the genomic structure of the RhD gene among apparently Rh-negative Chinese in Taiwan in this study. METHODS: Genomic DNA from 230 samples of apparently RhD-negative Chinese was studied using four polymerase chain reaction (PCR)-based RhD typing methods. These PCR methods amplified RHD and RHCE genes at exons 4, 5, 7 and 10. All nucleotides responsible for exofacial amino acid differences between RhD and RhCeEe peptides, including amino acids 169, 170, 172, 223, 226, 233, 238, 350, 353, and 354, were contained in these amplified DNA segments. Southern blot analysis using RHD cDNA fragments as probes was performed. RESULTS: According to the serological study, 155 samples (67.4%) were genuinely RhD-negative and 75 samples (32.6%) were of the Del phenotype. Successful amplifications for RHD sequences were possible in all 75 Del samples using four PCR methods. Apparently, all Del individuals carried an intact RHD gene. While 145 individuals of 155 genuinely Rh-negative (63.0% of apparently RhD-negative individuals) had total deletion of their RHD genes, 10 individuals (4.3% of apparently RhD-negative individuals) were shown to have a preserved 3' noncoding region of the RHD exon 10 and a gross deletion of RHD exons 4-10. CONCLUSIONS: Three classes of RhD-negative polymorphisms among Chinese in Taiwan were observed. These included Del with grossly intact RHD and weak RhD expression, genuinely RhD-negative with partial preservation of the RHD gene, and genuinely RhD-negative with total deletion of the RHD gene. A molecular study is warranted to clarify the mechanism responsible for the weak RHD gene expression in Del individuals. PMID- 9745153 TI - Different genotypes causing indiscernible patterns of A expression on A(el) red blood cells as visualized by scanning immunogold electron microscopy. AB - BACKGROUND AND OBJECTIVES: The published sequence of the weak. A subgroup Ael gene from Swedish individuals showed a G insertion in exon VII, causing a frameshift at codon 268 (the A1 gene has 353 codons). We wished to sequence exons VI and VII of two Norwegian Ael individuals and compare the expression of A substance on RBC from different Ael individuals. MATERIALS AND METHODS: Exon VI and VII were amplified by PCR, cloned in M13 and sequenced. A structure expression on Ael RBC was studied by the immunogold technique. RESULTS: In contrast to the Swedish Ael individuals, the two Norwegians had consensus A1 sequences in exon VI and VII. However, the patterns of A expression were indiscernible from the Swedish cases as visualized by immunogold labeling in SEM. In both cases, a few (1-2%) RBC were very strongly labeled, some were weakly labeled and the majority (95%) were unlabeled. CONCLUSION: Although some Ael individuals have an inserted nucleotide in exon VII of the ABO gene, others have consensus A1 sequence in exon VI and VII. However, we could not find any differences in phenotype by immunogold labeling in SEM. PMID- 9745155 TI - Characterization of a macaque anti-Rh17-like monoclonal antibody. AB - In order to produce macaque monoclonal antibodies (mAbs) against human red blood cell (RBC) antigens, macaques were immunized with human and gorilla RBCs and their spleen lymphocytes were fused with man-mouse heteromyeloma cells. One macaque-mouse heterohybridoma produced a macaque IgGx (Cvn2-4D5) which agglutinated all human RBCs but not rare human variants Dc-,D-, and Rhnull. Thus, Cyn2-4D5 exhibited RH17-like reactivity. The specificity of Cyn2-4D5 for RHCE encoded polypeptides was confirmed by specific immunoprecipitation of RhcE and RhCe polypeptides from human RBCs and the absence of immunoprecipitation of the RhD polypeptides extracted from D-RBCs. This study demonstrates that it is possible to produce macaque mAbs against human RBC blood group antigens. PMID- 9745156 TI - The ELO blood group polymorphism is located in the putative first extracellular loop of human erythrocyte band 3. AB - BACKGROUND AND OBJECTIVES: The low incidence red cell antigen ELO (700.51) has been excluded from 13 of 23 established blood group systems. Information relative to the Diego system has not been reported. To investigate a possible association between ELO and the gene controlling Diego blood group polymorphisms, we undertook molecular studies of AE1 (anion exchanger 1 gene). MATERIALS AND METHODS: DNA from a family segregating for ELO and from the original ELO proposita was amplified by PCR using intronic primers flanking exons 11-20 of AE1. Subsequently, single-strand conformational polymorphism (SSCP) analysis and DNA sequencing were conducted. RESULTS: We have observed an exon 12 SSCP in all ELO+ individuals tested. This SSCP is the result of a C-->T mutation in exon 12 of AE1 which leads to an Arg432-->Trp amino acid substitution in the putative first extracellular loop of band 3 and thereby accounts for the Diego blood group system polymorphism known as ELO. CONCLUSIONS: In light of these findings, the International Society of Blood Transfusion Working Party on Terminology for Red Cell Surface Antigens has designated ELO as a member of the Diego blood group system (010008 or D18) and rendered the previous numerical designation (700.51) obsolete. PMID- 9745157 TI - A simple strategy to look back on posttransfusion hepatitis B in a multitransfused patient. AB - BACKGROUND AND OBJECTIVES: In January 1996, a case of hepatitis B virus (HBV) seroconversion in a multitransfused patient was reported to the blood bank From March through October 1995, the patient had received 23 units of red cells and 30 units of pooled platelet concentrates, encompassing an exposure to a total of 200 whole blood donations. MATERIALS AND METHODS: In order to trace hepatitis B surface antigen (HBsAg)-negative but HBV-infectious blood donation(s), we tested samples of the donors obtained > or = 3 months after the implicated donations for anti-HBc (Corezyme EIA, Abbott). From 172/200 donors, archived samples of subsequent donations were available for this purpose. The remaining 28 donors were reinvited to the blood bank to obtain an additional blood sample for anti HBc testing. RESULTS: 1/200 follow-up donor samples was anti-HBc-positive. Retrospective testing of the implicated HBsAg-negative blood donation of this donor revealed anti-HBc-negative and HBV-DNA-positive results. The patient was transfused with the platelets of the HBV-infectious donation. On looking back, the other blood products prepared from this HBV-infectious donation caused posttransfusion HBV infection (PT-HBV) in 2 additional patients. CONCLUSION: Anti HBc testing on mainly archived follow-up samples of 200 donors implicated in PT HBV was a rapid, simple cost-effective and donor-friendly method to identify an infectious but HBsAg-negative, anti-HBc-negative and HBV-DNA PCR-positive blood donation. Routine anti-HBc screening would not have prevented this HBV transmission. PMID- 9745158 TI - Unsatisfactory detection of an in vivo haemolytic anti-vel by the gel test. AB - BACKGROUND AND OBJECTIVES: A patient experienced a severe haemolytic transfusion reaction. Neither the haemolytic property nor the specificity of the causative antibody had been sufficiently recognised when performing a microcolumn gel test. MATERIALS AND METHODS: Subsequent to the transfusion reaction, the serological property and specificity of the causative antibody were analysed. Tube and gel test methods were compared, as were various reagent red cell specimens and their constituents. RESULTS: A haemolytic anti-Vel was detected in the tube test. In contrast, the particular commercial gel test kit used did not reveal the haemolytic property or specificity of the antibody. Our experiments suggest that this was apparently due to the presence of EDTA in the low ionic strength saline solution of the test kit. CONCLUSIONS: In rare cases life-treatening haemolytic activity of an irregular blood group antibody may be undetected by a commericial microcolum gel test kit in which EDTA is a constituent. PMID- 9745159 TI - Tumor angiogenesis and metastasis: correlation in invasive renal cell carcinoma. AB - BACKGROUND: Experience suggests that tumor growth is dependent on angiogenesis. The intensity of angiogenesis in human cancer is reported to be predictive of the probability of metastasis in many types of cancer. The aims of this study were 1) to determine the relationship of microvessel density (MVD) in renal cell carcinoma to pathologic stage, and 2) to evaluate the role of MVD in metastasis. METHODS: Paraffin-embedded tumor specimens were reviewed from 34 unselected patients with RCC who had undergone surgery from 1986 to 1990 at Taichung Veterans General Hospital. The pathology findings and clinical records were reviewed to note relationships between pathologic stage and whether or not metastasis had occurred. Specimens were studied from 16 cases (eight Stage I cancers, five Stage II and three Stage III) without metastasis and from 18 cases (two Stage I, six Stage II, six Stage III and four Stage IV) in which metastasis later developed. Microvessels were highlighted by immunostaining endothelial cells for factor VIII-related antigen. Microvessels were counted in a x-400 field (0.1885 mm2/field) in the most active areas of neovascularization. RESULTS: The 16 patients without metastasis have survived for between 65 and 136 months (mean, 94.5 months), up to the present time. Of the 18 patients with metastasis, 15 died and three survived, with mean survivals of 42.8 months (range, 12-99 months). Mean overall MVD was 99.6 vessels; mean MVD was 98.5, 96.2, 109.3 and 90.0 in Stages I, II, III and IV tumors, respectively. Mean MVD was 99.3 in patients without metastasis and 99.9 in patients with metastasis. CONCLUSIONS: MVD does not correlate with pathologic stage and is of no prognostic significance in renal cell carcinoma. PMID- 9745160 TI - New one-week, low-dose triple therapy for the treatment of duodenal ulcer with Helicobacter pylori infection. AB - BACKGROUND: Antimicrobial therapy is the recommended treatment for duodenal ulcer associated with Helicobacter pylori infection. The eradication of bismuth-based triple therapy with bismuth subcitrate, metronidazole and amoxicillin is limited by low compliance, drug resistance and side-effects. Two-week proton pump inhibitor (PPI)-based triple therapy has a higher eradication rate but is costly. This study was designed to compare the efficacy, patient compliance and cost of short-term PPI-based triple therapy with those of bismuth-based triple therapy. METHODS: Ninety patients with active duodenal ulcer disease and H pylori infection, proven with the 13C-urea breath test and CLO test (Campylobacter-like organism test) were treated randomly in three therapeutic groups: Group A, DeNol 120 mg, amoxicillin 500 mg and metronidazole 250 mg four times a day orally for 14 days; Group B, omeprazole 20 mg plus clarithromycin 500 mg twice a day and amoxicillin 500 mg four times a day for 14 days; Group C, omeprazole 20 mg, clarithromycin 250 mg and metronidazole 500 mg twice a day for seven days. Nizatidine 150 mg twice a day was given continuously following the end of anti-H pylori therapy for each group. Two months later, endoscopy, the CLO test and 13C urea breath test were repeated to assess the eradication rate of H pylori and the ulcer-healing rate. Drug tolerance was evaluated by patients themselves by daily recording of any side-effects. RESULTS: Eighty-four patients completed the entire course of therapy and evaluation for H pylori infection. The H pylori eradication rates in Groups A, B and C were 75% (21/28), 93% (26/28) and 89% (25/28), respectively (p = 0.466). The ulcer healing rate was 86% (24/28) in Group A and 89% (25/28) in Groups B and C (p = 0.764). A total of 74 patients (88%) were free from symptoms at the end of the triple therapy. Symptom relief was faster in patients with PPI-based triple therapy (Groups B and C) (days 3 and 4) than for patients with bismuth-based triple therapy (day 5). The cost of Group C therapy was lower than that for Groups A and B. There were no major side-effects in any of the patients. CONCLUSIONS: One-week triple therapy with omeprazole, clarithromycin and metronidazole is highly effected for the eradication of H pylori. A therapeutic regime of one week's duration with lower cost, good compliance and mild side-effects may offer a good choice for treatment of duodenal ulcer associated with H pylori infection in clinical practice. PMID- 9745161 TI - Serum transferrin receptor concentration is not indicative of erythropoietic activity in chronic hemodialysis patients with poor response to recombinant human erythropoietin. AB - BACKGROUND: Serum transferrin receptor (sTfR) is a transmembrane glycoprotein derived from erythroid precursors in the bone marrow. Its concentration provides a quantitative measure of total erythropoietic activity and an indication of functional iron deficiency. This study was conducted to investigate whether sTfR is a useful index of erythropoietic activity in chronic hemodialysis patients with poor response to maintenance recombinant human erythropoietin (rHuEPO) therapy. METHODS: Using an enzyme-linked immunosorbent assay, sTfR concentration was measured in 67 uremic patients who had been on hemodialysis for a mean of 42 months (3-242 months). rHuEPO was administered three times a week to keep the hematocrit above 30%. Hemoglobin, red blood cell indices, serum ferritin, serum total iron binding capacity and unsaturated iron binding capacity were determined. Of the 67 patients, 35 who responded favorably to rHuEPO with hematocrits above 30% were categorized as Group I and 32 who did not attain the target hematocrit were categorized as Group II. As a control group, 31 healthy subjects were also investigated. RESULTS: The serum iron, ferritin, transferrin iron saturation, dialysis efficiency and nutritional state were not different between groups of hemodialysis patients. The mean sTfR concentration was 2.1 +/- 0.6 micrograms/ml (range, 1.15-3.53 micrograms/ml) in Group I patients, compared with 1.9 +/- 0.9 micrograms/ml (range, 1.03-2.65 micrograms/ml) in Group II. The difference was not significant. In addition, the mean sTfR concentration of 1.8 +/- 0.4 micrograms/ml (range, 0.86-2.76 micrograms/ml) in the healthy controls was not significantly different from Groups I and II. CONCLUSIONS: sTfR concentration cannot be used to distinguish good from poor rHuEPO responders among chronic hemodialysis patients who have elevated serum ferritin (> 300 micrograms/l) and transferrin iron saturation (> 25%) during the course of maintenance rHuEPO therapy. PMID- 9745162 TI - Vascularized iliac bone graft for treating avascular necrosis of the femoral head. AB - BACKGROUND: Nontraumatic avascular necrosis of the femoral head (ANFH) is a common disorder causing disability of the hip joint. The means for optimally treating this disease are still controversial. In this study we evaluated the relatively new technique of vascularized iliac bone grafting for treating ANFH. METHODS: From March 1990 to March 1992, 17 hips (15 patients) with ANFH were treated using the vascularized iliac bone grafting technique in our hospital. The patients included 12 men and three women, with an average age of 38 years. Steinberg's classification was used to categorize the severity of hip disease. The clinical results were classified as excellent, good, fair and poor according to symptoms, hip function and roentgenographic changes after surgery. Life-table analysis was applied to assess graft survival and the log rank test was used to compare statistical differences between the steroid-related and nonsteroid related groups. RESULTS: Cumulatively, 16 hips (14 patients), excluding one patient (one hip) lost to follow-up, were clinically evaluated for an average of 68 months. Among 12 hips at Steinberg stage II, eight progressed to stage IV, three to stage III and only one remained at stage II. Although most hips at stage II showed mild to moderate disease progression on plain radiography, 58% of the hips (7/12) that progressed to less than IVb showed good to excellent results. In the three hips at stage III, one progressed to stage IVa and two to stage V. In the two hips at stage IV, one remained at stage IV at final follow-up and the other was lost to follow-up. The steroid-related and nonsteroid-related groups did not differ with respect to clinical results. Both groups had 63% (5/8) good to excellent results (p > 0.05, log rank test). Only four hips were converted to prosthetic arthroplasties at final follow-up. Overall, 63% (10/16) of the hips had good to excellent results, 12% had fair results and 25% had poor results. Graft survival after seven years of follow-up was 63%. CONCLUSIONS: While treatment of ANFH still poses a challenge to orthopedic surgeons, the vascularized iliac bone grafting technique is a clinically acceptable option for treating the early stages of ANFH. PMID- 9745163 TI - Emergency medical resource use in Taipei city. AB - BACKGROUND: It is a worldwide trend to serve emergency patients at emergency department (EDs) staffed by board-certified emergency physicians (EPs). The question arises as to how many EPs are cost effective? This cannot be answered until utilization of emergency resources in the community is explored, so that appropriate plans for efficient ED use, staffing and training of EPs can be made. METHODS: A prospective randomized chart-review study was conducted from August 1, 1995, until May 31, 1996, in the EDs of eight large hospitals in Taipei City. These included Chung-Hsin Municipal Hospital (CHH), Chung-Shiaw Municipal Hospital (CSH), Ho-Ping Municipal Hospital (HPH), Jen-Ai Municipal (JAH), Mackay Memorial Hospital (MMH), Tri-Service General Hospital (TSGH), Veterans General Hospital-Taipei (VGH-T) and the Yang-Ming Municipal Hospital (YMH). RESULTS: Of 37,889 cases reviewed, the average daily ED census at each hospital was: VGH-T 198; TSGH 148; MMH 224; HPH 62; YMH 84; CHH 41; JAH 102; and CSH 139. At MMH, pediatric patients comprised 38.8% of emergency patients and the elderly comprised 49% of patients of VGH-T. The average age of patients was 56 years in VGH-T and 40 years in other hospitals. There were 71 (11.1%) pediatric patients and 3,087 (1.8%) adult patients taken to hospital by an emergency medical services (EMS) ambulance. A total of 176 pediatric and 2,230 (8.1%) adult patients belonged to triage categories 1 and 2. VGH-T had a much higher percentage of triage 1 and 2 patients (769, 13%) than the other hospitals. A total of 30.7% of pediatric and 17.4% of adult patients were classified as triage 4 (pseudoemergency). Patients requiring advanced life support, referral cases and patients requiring an intensive care unit (ICU) bed accounted for 1.1%, 0.9% and 0.6% of pediatric patients, and 6.8%, 3.4% and 1.8% of adult patients, respectively. These figures increased to 11.1%, 7.7% and 3.5%, respectively, in VGH-T. Among patients requiring ICU admission, 30% of pediatric and 53% of adult patients were not admitted on presentation to the ED. Of 10,099 children and 27,555 adults who were evaluated for case disposition, 651 (6.5%) and 4,061 (14.7%), respectively, were considered for admission. Among these 542 (83.3%) and 2,894 (71.3%), respectively, were admitted. CONCLUSIONS: VGH-T is a favorable training ground for EPs due to its high volume of adult and elderly, advanced life support eligible, ICU and high acuity patients. MMH is an ideal place for training in pediatric emergency medicine. Utilization of the 119 emergency response system is low. The number of patients with high acuity triage in Taipei City is low. However, Taipei City faces a shortage of acute care beds, especially ICU beds. PMID- 9745164 TI - Tympanoplasty for pars flaccida. AB - BACKGROUND: The formation of a retraction pocket of the pars flaccida remains a difficult problem for otologists to treat. It may lead to ossicular erosion and the development of a cholesteatoma, especially when the pocket is adherent to the malleus neck. We designed a new method of surgery for the treatment of small attic cholesteatomas. METHODS: From 1986 to 1996, 20 patients with a retraction pocket of the pars flaccida or a small attic cholesteatoma underwent surgery as described below. The complete lesion was removed after widening the posterior superior bony external ear canal wall and placing pieces of conchal cartilage (usually less than 10 pieces, according to their scutum defect) lateral to the malleus neck or incus. The temporalis fascia was then laid. RESULTS: The average follow-up period was 32 months. The paired t-test was used to compare the results of preoperative and postoperative air conduction and air-bone gap. The preoperative average air conduction was 31.17 dB and the air-bone gap was 15.09 dB. The postoperative average air conduction was 20.66 dB and the air-bone gap was 3.09 dB. The difference between preoperative and postoperative measurements was considered significant (p < 0.05). No recurrent retraction pockets or cholesteatomas were noted during follow-up. CONCLUSIONS: Tympanoplasty for correction of a retraction pocket of the pars flaccida can prevent further attic retraction and the development of cholesteatomas. Postoperative hearing results were also encouraging. PMID- 9745165 TI - Malignant fibrous histocytoma of the heart presenting as right ventricular outflow tract obstruction: a case report. AB - Primary malignant fibrous histocytoma (MFH) of the heart is extremely rare. Herein, we report the case of a 17-year-old woman with a primary MFH in the right ventricular outflow tract, presenting with increasing dyspnea on exertion. Wide excision of the tumor, including part of the pulmonary artery and pulmonary valve, was performed under cardiopulmonary bypass. Her postoperative recovery was uneventful. PMID- 9745166 TI - Pyomyositis in childhood: a case report. AB - Pyomyositis is a primary infection of skeletal muscle. We report the case of a previously healthy six-year-old who suffered from pyomyositis in the right lower back. He presented with lower back pain and low-grade fever for one week. After a series of laboratory and imaging studies, the diagnosis of right multifidus muscle pyomyositis with abscess formation was made. The patient recovered rapidly after incision and drainage therapy, accompanied by antibiotic treatment. Methicillin-resistant Staphylococcus aureus was cultured from the abscess discharge. It was strongly suspected that herbal medicines and common cold medication the patient had been prescribed before admission to our hospital produced a masking effect that delayed the diagnosis. PMID- 9745167 TI - Delayed postpartum hemorrhage in adenomyosis: a case report. AB - Adenomyosis of the uterus is a serious problem for women of reproductive age because of its possible consequence of infertility. We present the case of a woman who had adenomyosis of the uterus, a successful spontaneous pregnancy, and delayed and tenacious postpartum hemorrhage that did not respond to conservative treatment. The 26-year-old woman, gravida 1, para 1, suffered from fulminating vaginal bleeding and associated shock 20 days after the delivery of a 3,450-g female by Cesarean section. Conservative treatment included uterine compression and massage, blood transfusion, intravenous administration of high-dosage estrogen, oxytocin and sulprostone (prostaglandin E analogue), and gauze packing from the vagina into the uterine cavity. Despite treatment, the patient went into shock due to persistent vaginal bleeding. Emergency exploratoric assessment laparotomy was performed, followed by a stotal hysterectomy. Pathology revealed extensive adenomyosis of the uterus without other significant abnormality. The potential dangers of adenomyosis in pregnancy should not be overlooked when patients seek treatment for infertility. PMID- 9745168 TI - Choroidal involvement in Wegener's granulomatosis: a case report. AB - Wegener's granulomatosis is a necrotizing, granulomatous vasculitis. It usually causes sinusitis, pneumonitis and glomerulonephritis. The common ocular manifestations include conjunctivitis, scleritis, peripheral keratitis and orbital inflammation. We report the case of a 50-year-old woman with Wegener's granulomatosis and very severe ocular complications who underwent bilateral enucleation. The pathologic findings of the eyeballs revealed granulomatous necrotizing scleritis, perivasculitis and granulomatous choroiditis. The last, as far as we know, has not yet been reported. PMID- 9745169 TI - Universal antenatal screening for hepatitis B and immunisation of babies at risk. PMID- 9745170 TI - Enhanced surveillance of Mycobacterium bovis in humans. PMID- 9745171 TI - Acute suppurative otitis media: stage of exudation. PMID- 9745172 TI - The Onodi (sphenoethmoid) Cell. PMID- 9745173 TI - Granular cell tumor of the larynx. PMID- 9745175 TI - Congenital cholesteatoma of the middle ear. PMID- 9745174 TI - Oto-palatal-digital syndromes. PMID- 9745177 TI - Medical education and research: the impact of a changing healthcare system. PMID- 9745176 TI - How to kill managed care. PMID- 9745178 TI - Endolymphatic subarachnoid shunt failure caused by Silastic allergy. AB - During the period from 1964 through 1994, the endolymphatic subarachnoid shunt operation was initially successful in eliminating endolymphatic hydrops and the symptoms and findings it produces in 76% of 645 ears of patients with Meniere's disease. After initial success, lasting from five weeks to nine years, endolymphatic hydrops suddenly returned due to obstruction of the Silastic shunt tube in 11% of patients. In these cases, prompt revision can often restore an initial good result. Histologic and immunologic examination of the material surrounding and occluding the tubes showed an allergic response to the Silastic material in most instances. Efforts to eliminate this cause of failure using a tube of new design and different plastic material are described. PMID- 9745179 TI - A functional diagnosis of dysphagia using videoendoscopy. PMID- 9745180 TI - Giant pleomorphic adenoma of the parotid gland: case report and review of the literature. AB - Pleomorphic adenomas account for the majority of parotid masses, typically arising in the tail of the gland and enlarging slowly over time. The vast majority are 2 to 6 cm in size when resected. We report resection of the largest benign mixed tumor recorded in the modern English language literature. An 85-year old reclusive woman had a 20-year history of an enlarging right periauricular mass that had begun bleeding several days prior to admission. The patient ultimately underwent resection of the mass, which measured 26 cm in diameter, weighed 6.85 kg, and proved on pathologic examination to be a benign mixed tumor without malignant degeneration. The implications of this unusual case for the management of mixed tumors are discussed, and a review of the world literature on giant pleomorphic adenomas is presented. PMID- 9745181 TI - Thyroglossal duct cyst: the New York Eye and Ear Infirmary experience and a literature review. AB - Thyroglossal duct cysts often present in childhood but can also afflict the adult population. In 1920, Sistrunk described surgical management and advocated the removal of the central portion of the hyoid bone, following the cyst tract to the base of the tongue. This surgical technique has not changed since its description 60 years ago. In this paper, a retrospective review of 70 thyroglossal duct cyst excisions performed at the New York Eye and Ear Infirmary from 1988 through 1996 is presented. The patient population consisted of 43 females (61%) and 27 males (39%). The average age at presentation was 21.5 years, with a range of 18 months to 64 years. The most frequent presenting symptoms was a painless midline neck mass. Computed tomography (CT) was the most frequent imaging study performed. Sixty-four patients underwent a Sistrunk procedure while five patients had excision alone. One patient was diagnosed but lost to follow-up. All five patients who underwent simple cystectomy required a second procedure. One patient who underwent the Sistrunk operation required revision. Nine patients had postoperative complications, with recurrence being the most common. We present our experience over an eight-year period. PMID- 9745182 TI - Granular cell tumor of the larynx in a six-year-old child: case report and review of the literature. AB - Granular cell tumors (granular cell myoblastomas) are uncommon neoplasms in the adult population, occurring predominantly in the head and neck and most frequently in the tongue. Laryngeal presentations are unusual, and granular cell tumors of the larynx in children are extremely rare, with a total of 19 cases reported in the literature in children under the age of 17 years. We report an additional case of a laryngeal granular cell tumor, in a six-year-old boy, and discuss the clinical, histologic, ultrastructural and therapeutic aspects of these neoplasms. PMID- 9745184 TI - Traumatic neuromas of the head and neck. AB - An interesting case of a traumatic neuroma of the greater auricular nerve provides the impetus for a discussion of head and neck neuromas. Traumatic neuromas of the head and neck are relatively rare. Division of the greater auricular nerve during parotidectomy occasionally results in a traumatic neuroma. We report a case of a 73-year-old woman who presented with a traumatic neuroma nine years after undergoing superficial parotidectomy with dissection of the facial nerve for a mixed tumor. The patient had a 1.5 cm x 1.0 cm mass located below the old surgical site over the anteromedial border of the sternocleidomastoid muscle. The patient's past history was significant for Frey's syndrome, which is the result of abnormal neurologic growth. On first impression, the tumor was thought to be a recurrence of neoplastic disease; however, because of the evaluation, traumatic neuroma was suspected. An attempt at fine-needle aspiration of the mass was too painful to be carried out. At surgery, a whitish tumor was excised which, on final pathologic examination, revealed traumatic neuroma. The surgical literature is reviewed and the subject of head and neck neuromas, including their evaluation and management, is thoroughly discussed. Knowledge of this possible diagnosis may spare the patient and the surgeon needless worry, as well as unnecessary procedures, once tumor recurrence has been ruled out. PMID- 9745183 TI - Cochlear neuronal loss are determined by use of a neurofilament protein antibody in cases of bilateral profound deafness. AB - The temporal bones of two patients with profound bilateral deafness from infancy were studied immunohistochemically, using a neurofilament protein antibody to detect the cochlear neuronal elements. One patient exhibited Mondini dysplasia of the inner ear, with the organ of Corti almost completely deteriorated. The other patient is the first reported case involving complete aplasia of the organ of Corti in all turns. In both cases, the immunohistochemical staining clearly revealed a severe reduction in the number of afferent neurons, such as dendrites, spiral ganglion cells and cochlear axons. The number of efferent spiral bundles in the osseous spiral lamina and intraganglionic portion also decreased in parallel with the reduction in the number of cochlear afferent neurons. Our results are inconsistent with previously reported cases of presbycusis and acquired deafness induced by the measles virus, in which efferent neurons were preserved while afferent neurons degenerated. The loss of both the efferent and afferent neurons might be characteristics of congenital deafness. PMID- 9745185 TI - The role of powered instrumentation in the surgical treatment of allergic fungal sinusitis. AB - Allergic fungal sinusitis is a chronic disorder that is being more frequently recognized by otolaryngologists. It is a recurrent illness characterized by frequent exacerbations, and requires aggressive medical and surgical treatment. When surgical therapy is employed, it is necessary to ensure adequate debridement and removal of edematous tissue. We have been using powered dissection as our primary method in sinus surgery over the past three year. We have treated 11 patients with allergic fungal sinusitis, and find powered instrumentation to be very effective in removing the polypoid tissue from the nose and sinuses, and in providing a clear surgical field. The procedure can be performed safely with minimal trauma to normal tissue. We believe that the use of powered dissection greatly enhances the comprehensive treatment of allergic fungal sinusitis. PMID- 9745186 TI - Top ten errors revisited. PMID- 9745187 TI - Advertisement from Mentor Ophthalmics Inc. and concerns regarding the use of Polytef. PMID- 9745188 TI - Effect of interferon therapy on heart rate and sympathetic nervous system. AB - We investigated the hypothesis that interferon-induced hyperadrenergic state causes cardiovascular events. Thirteen patients with chronic hepatitis C who were scheduled to undergo interferon therapy were randomly selected for this study. Heart rate was monitored and urinary levels of catecholamines were measured before, on the 1st day, and after 1 week and 4 weeks of treatment. On the 1st day, 24-hr average heart rate and hourly average heart rate were significantly higher than those before treatment, although they returned to baseline levels after 1 week and 4 weeks. However, urinary levels of cathecholamines did not change significantly on the 1st day of interferon therapy. Although the direct evidence that interferon stimulates sympathetic nervous system could not be obtained, interferon elicited tachycardic response and may have increased the risk of arrhythmias of the 1st day of the treatment. PMID- 9745189 TI - Primary localization amyloidosis of the sublingual gland. AB - We here in present a very rare case of primary localized amyloidosis with amyloid A protein of the sublingual gland. It presented a tumorous appearance on the left oral floor. Pretreatment with potassium premanganate made biopsy specimens unstained by Congo red. Immunohistochemical staining for AA protein was positive. Systemic amyloidosis was ruled out based on clinical and laboratory examinations. The gastric and the labial salivary glands biopsy showed no amyloid deposits. As far as we know, this is the first case of primary localized amyloidosis with amyloid A protein of the oral cavity, and tumor-formed amyloidosis of the sublingual gland. PMID- 9745190 TI - [Microbial etiology of mild and moderate pelvic inflammatory disease]. AB - Pelvic inflammatory disease (PID) is one of the most severe complications of sexually transmitted disease (STD). It can be due to the ascending of normal endogenous microorganisms of the female genital tract or the infection by microorganisms related to STD as Chlamydia trachomatis and Neisseria gonorrhoeae. PID leads to serious gynecoobstetric consequences as infertility and ectopic pregnancy. Clinicians face the problem of knowing the ethiology of PID in order to treat appropriatly patients with this clinical diagnosis. So that, this work pretends to establish what kind of microorganisms are implicated in PID. A proper isolation and identification of microorganisms achieved by culture of lower genital tract samples from endocervix, endometrium and peritoneal fluid, leading to a betther, specific and proper treatment of this disease. PMID- 9745191 TI - [Efficacy of isosorbide in aerosol form in the management of hypertensive crisis in severe preeclampsia]. AB - In this assay, we evaluated the utility of isosorbide given by a sprayer in the management of hypertensive crisis of severe preeclampsia. 36 pregnant women with severe preeclampsia received fluid therapy and were randomly dividend in two groups of 18 patients each. Group A, received isosorbide in spray 1.25 mg when admitted and a second dose 10 minutes after if mean arterial blood pressure decreased less than 15%. and group B, in whom 4 g of magnesium sulphate was infused in one hour and then 1 g each hour for a maximum of five hours. In all patients blood pressure, proteinuria, fetal and maternal heart rate and Apgar score at minute and five minutes were obtained. In Group A 13 patients had a significative blood pressure reduction with one application and 5 needed a second one (p < 0.002). fetal (p < 0.005), and maternal (p < 0.005) heart rate also had a significative reduction. Whereas three patients in Group B did not respond and the rest had a poor blood pressure control (p > 0.05) with no changes in fetal and maternal heart rate. No patient developed eclampsia. When compared both groups, there were a significative difference for blood pressure (p < 0.005), fetal heart rate (p < 0.002), maternal heart rate (p < 0.05) and Apgar at minute (p < 0.01) in isosorbide's group. Our data suggest that isosorbide given by a sprayer is effective and safer in the management of the hypertensive crisis of severe preeclampsia. PMID- 9745192 TI - [Prognostic importance of controlled ovarian hyperstimulation previous to therapeutic laparoscopy of infertile patients]. AB - The use of the laparoscopic approach in the initial evaluation of the infertile couple without obvious disease is controversial; for these reason some centers has been done the initial treatment with ovarian stimulation (OS). The present article analyze two groups, the first for patients with OS for 4 cycles and laparoscopic approach after that and the second group with initial laparoscopy and medical treatment if they need it. Both groups has a OS after the laparoscopic procedure. The analysis of the prognosis of the OS before the laparoscopy and the impact in the reproductive outcome is done. The results shows that the OS does not have deleterious effects of the pregnancy rates if it is used before the procedure. However, the pregnancy are present early in the second group P < 0.05. In conclusion, in infertile woman without obvious disease that indicate the laparoscopy, the initial use of OS before the laparoscopy does not have a deleterious impact on the pregnancy rates. PMID- 9745193 TI - [Trying vaginal delivery in 1000 patients with previous cesarean section in the Antiguo Hospital Civil de Guadalajara]. AB - Because of the main justification for practicing a cesarean section is due to a previous cesarean and the rasing rates frequency of this operation, we concluded a descriptive and prospective investigation in order to analize the factibility and security of vaginal delivery after one cesarean section. We include 1000 patients with a past history of one previous cesarean section and with the following main characteristics: normal evaluation of the actual pregnancy and a gestational age of at least 36 weeks of pregnancy, no pelvis stenosis and a normal fetal status. The management were expectant and or with the use of oxitocin, prostaglandin PGEJ, uterionhibition and or amnioinfusion according to medical indication, 679 (67.9%) patients had a vaginal delivery; one ruterine rupture (0.001 x 1000) happened (the place of the rupture were not in the scar of the previous cesarean); two uterine dehicence (0.002 x 1000) of the previous uterine scar; one of this require laparotomy and sture of the dehiscence scar and the other one only require observation. We had two intrapartum fetal dead (0.002 x 1000) on due to the uterine rupture and the other one because of a taquisitolia not corrected by betamimetics. The factibility and security of vaginal delivery after one previous cesarean section is a logical and reasonable strategy in order to decrease the actual high rates of cesarean section. Whenever we try a viginal delivery in a patients with one previous cesarean is imperative to keep in mind that if something is not going well during the attempts we must repeat another cesarean. PMID- 9745194 TI - [Cytologic correlation between the Bethesda system and colposcopic biopsy]. AB - Bethesda system is useful in the citologic diagnosis of premalign diseases of cervix. It was determined the correlation between citologic report using Bethesda system and combined colposcopic index (CCI) or Reid, to diagnose premalign diseases of cervix by histologic diagnostic of punch biopsy by colposcopy. A total of 118 patients come to the colposcopy clinic having smear anormal citology to cervical intraepithelial neoplasia (CIN), only o associated to HPV. It was taken to all of them a new cervical smear, colposcopy and punch biopsy. We analized the findings for citology with Bethesda system and colpospic with CCI of Reid, using histologic diagnosis of punch biopsy by colposcopy. Diagnosis correlation between Bethesda system and the punch biopsy by colposcopy was 98.5% for 82 patients with low grade squamous intraepithelial lesions with a concordance of 100% for HPV and 97% for CNI I (p < 0.05). In the other hand 92% from 36 patients classified as high grade squamous intraepithelial lesions with a concordance of 84% to CIN II and 100% to CIN III (p < .05) Sensibility of 93%, especifidad and positive predictive value of 96% and negative predictive value of 98%. We conclude correlation between citologic report with Bethesda system and the Combined Colposcopic Index of Reid which are effective to diagnose premalign disease of cervix. PMID- 9745195 TI - [Classification of the characteristics of cesarean section at the Mexican Ministry of Health during the period of 1990-95]. AB - This is an analysis of the characteristics of cesarean section operations within the Ministry of Health (SSA) and the implications in relation to maternal and perinatal mortality during the five year period from 1990 to 1995. A descriptive study was undertaken based on cesarean section information obtained from the Statewide (Reporting) System of Basic Information (SEIR) and based on maternal and perinatal mortality information provided by the Ministry's Internal Hospital Committees from 1990 to 1995. This information was analyzed as a chronological series in order to compare the changes in the annual cesarean rates at both national and statewide levels. The Pearson test was used to determine the correlation between the frequency of cesarean sections and maternal and perinatal mortality rates during the same period within a 95 percent confidence interval (p < 0.05). The frequency of cesarean sections was shown to have increased within the SSA hospital system as whole and when considering each state separately. A positive and statistically significant correlation was observed between the frequency of cesarean sections and maternal mortality rates, while the increase in cesarean rates demonstrated no relationship with perinatal mortality rates. The increase in cesarean section rates in the Ministry of Health during the period analyzed exceeded any potential benefits and could have contributed to the increase in maternal morbidity and morbility as well as hospital cost. This increase is worrisome due to the projected tendency for the rates to continue to increase in the next few years if quick and concrete actions are not taken in order to decrease these rates. PMID- 9745196 TI - [Structural characteristics of prolactin, growth hormone and their receptors as determinants of biological actions]. AB - The pituitary hormones prolactin and growth hormone are structurally related. Both hormones exist in the circulation in several molecular forms, differing in aminoacid sequences, posttranslational modifications and fragments produced by proteolytic cleavage. Heterogencity may produce a diversity of inmunological and biological actions. It has been suggested that each of this forms may be a isohormone with a different physiological role. The predominance of one of them in serun could account for the complex and often contradictory actions of the hormones. In addition receptors also have structural homology and so the possibility exist that these hormones share binding affinity to the receptors and can produce endocrinological problems in some special conditions. PMID- 9745197 TI - [Antithrombin-III in preeclampsia-eclampsia. Pilot study]. AB - The functional levels of AT-III were determined to the following groups: A. Eleven healthy non pregnant women B. Thirteen healthy pregnant women (third trimester). C. Six preeclamptic patients. D. Five patients with eclampsia and/or HELP syndrome. The results were as follows: [table: see text] A different grade of DIC may explain the low activity of AT-III in preeclampsia and a more severe coagulation disorder in eclampsia and HELP syndrome. Our preliminary results encourage other prospective studies including larger populations to determine its usefulness as early diagnostic test and severity marker of the disease. PMID- 9745198 TI - [Integral treatment of severe ovarian hyperstimulation syndrome by means of autotransfusion of ascitic fluid and intravenous infusion of albumin]. AB - Severe ovarian hyperstimulation syndrome (OHSS) is a potential life-threatening condition relationated with ovulation induction. It affects multiple systems. Little is known about it's pathophysiology. The treatment available consists in the correction of fluid, electrolyte and hematologic imbalances. In other hand, is mandatory the prevention of embolic phenomena. Ascitic fluid aspiration result in dramatic improvement of symptoms. The purpose of this study was to assess the effect of autotransfusion of ascitic fluid obtained by paracentesis and the intravenous infusion of albumin for the treatment of severe from of the OHSS. PMID- 9745199 TI - APECED: a monogenic autoimmune disease providing new clues to self-tolerance. PMID- 9745200 TI - Gene therapy in osteoarticular diseases: where are we? PMID- 9745201 TI - Rocking the foundations of solid tumor growth by attacking the tumor's blood supply. PMID- 9745202 TI - A very high level of crossreactivity is an essential feature of the T-cell receptor. PMID- 9745203 TI - Tumors express both unique TSTA and crossprotective 44 kDa oncofetal antigen. PMID- 9745204 TI - Dialogue between the CNS and the immune system in lymphoid organs. PMID- 9745205 TI - Immunology of the tonsils. PMID- 9745206 TI - Human marginal-zone B cells. PMID- 9745207 TI - Langerhans cells, keratinocytes, nitric oxide and psoriasis. PMID- 9745208 TI - Crossreactivity of the T-cell receptor. PMID- 9745209 TI - Alternative non-steroidal anti-inflammatory therapy for asthma. PMID- 9745210 TI - Re-emergence of Vibrio cholerae serogroup O139 during June-August, 1997 in Yavatmal (Maharashtra). AB - A clone of V. cholerae serogroup O139 which emerged as a novel epidemic strain, was reported from this region in 1993 as from many other parts of India and adjoining countries. The decline in the isolation rate of this organism in subsequent years was followed by a sudden increase in 1997, this requires careful monitoring. PMID- 9745211 TI - Oxidative stress response in Shigella & nonpathogenic gut bacteria. AB - The effect of oxidative stress in the form of exogenous H2O2 on the survival of four species of Shigella and two nonpathogenic Gram negative gut bacteria and the role of catalase as an antioxidant enzyme, neutralizing the effect of H2O2 were examined. A significant decrease in the number of colony forming units (CFUs) after exposure to exogenous H2O2 (122 +/- 37), compared to control bacteria (218 +/- 63, P < 0.001) was observed. There was an induction of catalase activity after exposure to exogenous H2O2 and the specific activity of catalase in H2O2 exposed bacteria was significantly increased (2.88 +/- 1.25), compared to control bacteria (1.5 +/- 0.44; P < 0.05). A direct correlation was observed between the decrease in bacterial counts and increase in catalase activity after exposure of H2O2 (regression coefficient (0.56). Gut bacteria appear to be susceptible to oxidative stress and inducible catalase activity may form an important part of the antioxidant defence mechanism against oxidative stress. PMID- 9745212 TI - Evaluation of serum neutralizing antibodies to bovine coronavirus in cows & their calves using Hmlu-1 cells. AB - Between 1992 and 1993, 75 paired serum samples from Holstein dairy cows and their calves were collected from Aomori, Tochigi and Okinawa Prefectures, and the neutralizing antibody titres to bovine coronavirus (BCV) were determined using hamster lung (Hmlu)-1 cells. The anti-BCV antibody positive rate in the maternal serum samples was significantly lower (P < 0.001) in Okinawa (72%) than in Aomori (100%) or Tochigi (100%). The geometric mean tire (GMT) of anti-BCV neutralizing antibody was also significantly lower (P < 0.05) in maternal sera from Okinawa (89) than that of Aomori (229) or Tochigi (264). The anti-BCV neutralizing antibody titres in the sera of calves which had ingested the colostrum, significantly correlated with the antibody concentration of the maternal serum samples (P < 0.05). These results suggest an extensive BCV infection among the dairy cattle in these prefectures, with a varied pattern of distribution between the prefectures. Anti-BCV neutralizing antibody in the sera of newborn calves appeared to be transferred from their dams through colostrum. PMID- 9745213 TI - Prevalence of fragile X(A) syndrome in mentally retarded children at a genetics referral centre in Delhi, India. AB - The usefulness of the clinical score based on Turner 5-trait scale prior to undertaking cytogenetic or molecular tests for the diagnosis of the fragile X(A) syndrome was evaluated. Mean clinical score in fragile X positive patients was significantly higher than in fragile X negative patients (7.06 +/- 1.85 vs 2.98 +/- 1.6, P < 0.0001). Of 1206 children with mental retardation 360 (29.8%) boys fulfilled defined clinical criteria to be screened for fragile X syndrome by chromosomal studies. Twenty three (6.38%) of them were found to be positive for fragile X syndrome using cytogenetic techniques. Molecular confirmation in 21 affected boys (two were lost to follow up) showed full mutation in 19 (5.27%). Two patients showed a normal 5.2 kb band on southern blot. This frequency (5.27%) of fragile X(A) patients among children with non-specific mental retardation is comparable to the results of studies in the West. Routine use of the clinical score, and the selection of patients with a score > or = 5 for diagnostic tests would reduce the laboratory load, especially in countries with limited facilities. PMID- 9745214 TI - Effect of DEHP [di-(2-ethyl hexyl) phthalate] on lipid peroxidation in liver in rats and in primary cultures of rat hepatocytes. AB - The effect of DEHP [di-(2-ethly hexyl) phthalate] on lipid peroxidation in the liver in rats and in primary cultures of rat hepatocytes incubated with it was studied. The doses of DEHP used in this study corresponded to the low levels of this substance leaching into blood stored in DEHP plasticised PVC bags. Increased activity of superoxide dismutase (SOD) and catalase, increased concentration of malondialdehyde (MDA) and conjugated dienes and decrease in the concentration of glutathione and vitamin E have been observed in the liver of rats administered DEHP. Primary cultures of rat hepatocytes incubated with DEHP also showed increase in the activity of these enzymes, increase in the concentration of MDA and decrease in vitamin E. These results indicate that DEHP promotes lipid peroxidation. Incorporation of vitamin E along with DEHP into the culture medium containing hepatocytes counteracted these effects. PMID- 9745215 TI - Flexible dose open trial of Vijayasar in cases of newly-diagnosed non-insulin dependent diabetes mellitus. Indian Council of Medical Research (ICMR), Collaborating Centres, New Delhi. AB - A flexible dose open trial was undertaken in four centres in India to evaluate the efficacy of an Ayurvedic drug Vijayasar (Pterocarpus marsupium) in the treatment of newly-diagnosed or untreated non-insulin dependent diabetes mellitus (NIDDM). By 12 wk, control of blood glucose (both fasting and postprandial levels) had been attained in 67 (69%) of 97 patients studied, and the dose on which control was attained was 2 g of the extract in about 73 per cent of the patients, 3 g in about 16 per cent and 4 g in 10 per cent of the patients. Four patients had to be withdrawn from treatment due to excessively high postprandial blood glucose levels. Among the 93 patients who completed 12 wk of treatment, both the fasting and postprandial blood glucose levels fell significantly (P < 0.001), by 32 and 45 mg/dl at 12 wk from the initial means of 151 and 216 mg/dl respectively. Mean HbA1c decreased significantly (P < 0.001) to 9.4 per cent at 12 wk from the initial mean of 9.8 per cent. No significant change was observed in the mean levels of lipids. Other laboratory parameters remained stable during the designated treatment period of 12 wk. Also, no side-effects were reported. Hence, it is concluded that Vijayasar is useful in the treatment of newly diagnosed or untreated mild NIDDM patients. PMID- 9745216 TI - [Clinical analysis of patients with sepsis--comparison between underlying diseases]. AB - We evaluated the clinical data in 83 patients with sepsis, which was diagnosed by both Bone's definition of sepsis and positive isolates from blood culture, according to their underlying diseases. This study enrolled a total of 117 septic episodes in 83 patients (57 males and 26 females, mean age: 52.0 years). We classified 3 groups, including hematological malignancies (46 patients, 72 episodes), solid malignant tumors (23 patients, 25 episodes) and non-malignancies (14 patients, 20 episodes), by the underlying diseases. Of the total number of isolates from blood culture, 53.0% were single gram-positive bacteria, 33.3% were single gram-negative bacteria, 7.7% were single fungus and 6.0% were polymicrobial organisms. In addition, coagulase negative staphylococci was isolated most often in patients with hematological malignancies. Sepsis was often caused by infectious focuses of hemorrhoid, stomatitis or intravenous catheter in patients with hematological malignancies, by pneumonia in patients with solid malignant tumors and by urinary tract infection in patients with non malignancies. Mortality of sepsis in patients with solid malignant tumors (48%) was highest in 3 groups. Septic patients, who were complicated with shock and/or DIC, has poor prognosis in all groups. Serum albumin level was significantly lower in dead patients than patients who survived. These results suggest that clinical features may be different according to the underlying diseases of patients with sepsis. PMID- 9745217 TI - [Analysis of transmission of Burkholderia cepacia isolates in an intrahospital by randomly amplified polymorphic DNA-PCR method]. AB - Strains of Burkholderia cepacia isolated in our hospital from November 1995 to September 1996 were classified with randomly amplified polymorphic DNA-PCR (RAPD PCR) and conventional biochemical tests (ID test.NF-18, API20NE, and Neg Combo 4J kit), and intrahospital isolates of B. cepacia were analysed. During the period 28 strains from inpatients and 2 from medical apparatus were isolated. Twenty four of 28 (85.7%) were from sputum. In 1996 from January to February, 20 strains were detected from 8 inpatients, and two strains were from the nebulizers at the Trauma and Critical Care Center (TCC). With typing of B. cepacia by conventional methods no epidemiological relations among isolates were found. However, DNA patterns of original isolates from the nebulizers at TCC by RAPD-PCR were identical with those of isolates in sputa from patients in other wards who had stayed at TCC, indicating that TCC was an initial source of transmission and the strain was transmitted with the patients to the wards. These results suggest that RAPD-PCR method might be a useful tool to analyse an epidemiological survey for intrahospital transmission of isolate. PMID- 9745218 TI - [Studies for dynamics of reverse transcriptase inhibiting antibody in sera from HIV-1 infected individuals]. AB - Antibodies inhibiting human immunodeficiency virus type 1 (HIV-1) reverse transcriptase activity (RTI-antibody), Binding inhibition antibody (BI-antibody) and polymerization inhibition antibody (PI-antibody) were investigated for their ability to inhibiting RT activity in 248 HIV-1 infected individuals and 99 healthy individuals. In BI-antibody, high titer samples were determined more in than in RTI- and PI-antibodies. No significance was indicated between AC, ARC and AIDS is any antibody, however, progression from AC to AIDS was poled to high titer and low titer in RTI- and BI-antibodies. Moreover, time course of each antibody levels in the same infected individuals were resulted in no change, going up or down through all the experimental term, though all samples were collected in AC. These results were suggested that the determination factor of each stage in HIV progression would be multiple, and that the various dynamics of RTI-, BI- and PI-antibodies in the same infected individuals might be caused in the term from HIV infection to AIDS progression, prognosis or appearing of the drug resistant strain but stages of the disease. PMID- 9745219 TI - [An epidemiological investigation for MRSA and PRSP in Kinki area. Kinki Infection Working Group]. AB - Recent trends in the development of resistance of the Staphylococcus aureus and Streptococcus pneumoniae to antibiotics were investigated, using a questionnaire delivered to participants at a meeting of the Kinki District Society of Infections. Methicillin-resistant Staphylococcus aureus (MRSA) accounted for 55.4% of all isolated S. aureus, and more than 80% of MRSA was detected within hospitals. In outpatients, MRSA was often detected in pus, while in hospitalized patients, MRSA was often detected in sputum. Further, MRSA was accompanied by some other organisms (most frequently Pseudomonas aeruginosa) in 64.7% of MRSA positive patients. The sensitivity of MRSA to vancomycin (VCM) was 100%, to sulfamethoxazole-trimethoprim (ST) 99.2%, and to arbekacin, 98.6%. In contrast, Penicillin-resistant Streptococcus pneumoniae (PRSP) accounted for 42.4% of all isolates of Streptococcus pneumoniae. About 50% of PRSP was detected in out patients. For both hospitalized patients and outpatients, PRSP was most frequently detected in sputum. PRSP was accompanied by some other organisms (most frequently Haemophilus influenzae) in 49.3% of PRSP positive patients, PRSP had high sensitivity to cephems, carbapenems and VCM. PMID- 9745220 TI - [A familial outbreak of verotoxin-producing Escherichia coli O103:H2 infection in which a calf was the suspected infectious source]. AB - A familial outbreak of Verotoxin-producing Escherichia coli (VTEC) infection occurred in July 1996 in AKITA prefecture. Four VTEC strains harboring VT-1 and eaeA genes were isolated from three patients and a calf, breeding farm for which was located as close as 4 meters from the house where the patients lived in. All of the 4 VTEC isolates were serotyped as O63:H2 using commercially available sera kits. However, a patient isolate, EC-281, was serotyped as 0103:H2 at the International Escherichia and Klebsiella Centre. Titration and absorption tests using rabbit antisera raised against EC-281 confirmed that the serogroup of the remaining 3-VTEC isolates was also O103. Epidemiological characteristics including plasmid profile, antibiotic susceptibility patterns and pulsed-field gel electrophoresis patterns of the 4 VTEC isolates were completely the same, indicating that these isolates originated from a common source. These findings in conjunction with the results of epidemiological survey conducted by the Health Center suggested that a possible infectious source for this outbreak is the calf. Our present results strengthen the significance of calf as an infectious source of VTEC infection. PMID- 9745222 TI - [A number of Vibrio cholerae non-O1 isolated from aquatic environments]. AB - To estimated existence of Vibrio cholerae non-O1 in aquatic environments, the organisms isolated from river, estuary and sea water. V. cholerae non-O1 isolated form midstream and estuary water could be counted from 1.6 to 2400 CFU/100 ml by the membrane filtrated method (MF). V. cholerae non-O1 existed in midstream water more than in estuary water. However, the isolated organisms from estuary rate by MF (37.5%) was lower than it by alkaline peptone enrichment medium method (AP) (75.0%), as a result of halophilic bacteria grow an selected medium of MF. And the number of V. cholerae non-O1 isolated from aquatic environment did not correlate environmental parameters. The number of V. cholerae non-O1 isolated from river water varied, it suggested that the organism collectively adhere a floating matter. V. cholerae non-O1 was not detected in 500 ml sea water by AP and MF method. These results conclude that V. cholerae non-O1 exist in river water more than in sea. PMID- 9745221 TI - [Possibility of prevention of herpes zoster by use of varicella vaccine]. AB - It has been considered that a decline in specific cell-mediated immunity (CMI) for the varicella-zoster virus (VZV) could be responsible for a high incidence of herpes zoster in the elderly. If the strength of CMI for VZV could be increased by immunization of the elderly with a varicella vaccine, herpes zoster might be preventable. We compared the CMI for VZV (using a lymphoproliferative assay and a varicella skin test) and VZV-IgG antibodies in serum before and after 2-3 months of vaccination in 15 subjects more than 40 years old. When the CMI for VZV was measured by the lymphoproliferative assay, a stimulation index (SI) of more than 2.0 was estimated to be positive in this study. The SIs (mean +/- SD) before and after the vaccination were 2.7 +/- 1.8 and 2.7 +/- 1.9, respectively, and no significant difference was noted. On the other hand, the diameter of erythema in the varicella skin test after the vaccination became larger than that before the vaccination in the 10 of 13 subjects. In addition, serum VZV-IgG antibodies increased after vaccination in 6 of 14 subjects. There were no obvious reasons for the discrepancy in the results of the lymphoproliferative assay and the varicella skin test. However, because of the poor response indicated by the assay, only one vaccination for the elderly might not be enough to increase the CMI for VZV. The appropriate age for vaccination should also be considered. Lastly, further investigation of the CMI for VZV before and after vaccination on larger scale is required. PMID- 9745223 TI - [Reverting effect of cysteine on the suppression by glucose or dexamethasone of anti-Candida activity of human neutrophils]. AB - Amino acid mixture prescribed for an hyperalimentation solution (PN-twin) diminishes suppression of anti-Candida activity of neutrophils as reported previously (Tansho, T, et al. J. Jpn. Assoc. Infect. Dis. 70: 463-469). The aim of this study was to identify the active principle in the amino acid mixture and to examine its action mechanism. Amino acid mixture (PN-twin) containing 23 amino acids neutralized the suppression of anti-Candida activity of human neutrophils by 1.0% of glucose. These amino acids were divided to several groups by their structure and effects of the groups on the suppressed anti-Candida activity neutrophils were examined. In all groups tested, amino acids containing cystein and methionine clearly neutralized the suppression, especially cysteine at the concentration more than 20 micrograms/ml significantly recovered the anti-Candida activity of neutrophils which was suppressed in the presence of 1% glucose or 10( 6) M dexamethasone. Correspondingly, cysteine augmented production of lactoferrin by stimulated neutrophils; which functions as a major effector molecule in growth inhibition of Candida by neutrophils. These results suggest that cysteine in alimentation solution augments anti-Candida defense mechanisms through recovery of neutrophil function. PMID- 9745224 TI - [Identification of Escherichia coli O157 antigen by polymerase chain reaction]. AB - An assay was developed for the specific detection of Escherichia coli O157 using PCR, because O serological cross-reactivities have been reported between E. coli O157 and some E. coli, other bacterial species. PCR amplification of E. coli O157 rfbE (Ec O157:H7) gene that is necessary for the expression of the O157 antigen, was performed for the identification of E. coli O157. All Shiga toxin-producing Escherichia coli (STEC) O157:H7 and O157:H, non-STEC O157 strains were positive, and other non-O157 E. coli strains were negative by PCR. All tested strains of other bacterial species, like Salmonella O30 and Citrobacter freundii which gave positive results with O157 detection kits, were negative by PCR. It is recommended that PCR amplification of O157 rfbE gene is one of the most specific method for E. coli O157 identification. PMID- 9745225 TI - [Mycoplasma pneumoniae detection from throat swab by two-step polymerase chain reaction]. AB - Two-step polymerase chain reaction (PCR) with primers designated against 16S rRNA gene of Mycoplasma pneumoniae for diagnosis of infection was evaluated in comparison with the conventional single-step PCR and culture methods. The two step PCR method showed specific amplification of M. pneumoniae DNA and higher sensitivity (1.5 fg/assay) than the single-step PCR method. With the two-step PCR method, 76 of 322 throat swabs (23.6%) from patients with acute respiratory complaints gave positive results whereas 20.2% were positive in the culture method. Seven of 13 samples which were negative in the single-step PCR method but positive in either serological or the culture method showed positive results by the two-step PCR method. In addition, 5 samples which were weakly positive in the single-step PCR method showed distinctly positive results in the two-step PCR. These results indicate that the two-step PCR method is a useful tool for detection of M. pneumoniae in clinical specimens, although it requires a relatively sophisticated in technique. PMID- 9745226 TI - [Outbreak of aseptic meningitis in Iwamizawa, 1997, caused by echovirus 30]. AB - To investigate the clinical character of an outbreak of aseptic meningitis in Iwamizawa 1997 caused by echovirus 30, and to investigate the spreading of the outbreak, we analyzed clinical character of 75 hospitalized patients in our hospital, and mapped the patients' distribution in Iwamizawa City each week. We detected in our hospital an epidemic outbreak of acute enteroviral meningitis caused by echovirus type 30 in Iwamizawa, from September to December, 1997. Regarding the patients, there was little prevalence in males, with an average age of 6 years and a range of 0 to 13 years of age. The most constant symptoms were three major one such as headache (90%), fever up (89%), vomiting/nausea (87%), sometimes sorethroat (30%) and abdominal pain (15%). One case had a febrile convulsion temporally, and two cases had acute meningoencephalopathy and- encephalitis. In the cereblospinal fluid (CSF), we found no predominance of mononuclear cell (MNC) (58%) in the differential cell count. The mean of the peak of CSF cell counts was 654/3. White blood cell (WBC) was 8940/microliters, and CRP 1.4 mg/dl. None of them was detected in the bacterial culture of the CSF. Viral cultures were performed on CSF in 26 cases. Echovirus type 30 was isolated in 4 cases of hospitalized patients, and in one case with meningismus without pleocytosis. The beginning of the outbreak was observed in two kindergarten and one elementary school side by side. The peak of the whole outbreak was detected in the 3rd to 6th week, however the school spreading peak was detected in the 3rd and 4th week, and spreading was going in the whole city. PMID- 9745227 TI - Increased production of interleukin-10 by human blood monocytes stimulated with Mycobacterium avium-intracellulare complex. AB - Macrophages produce various cytokines in response to mycobacteria, including interleukin 10 (IL-10) and tumor necrosis factor alpha (TNF-alpha). IL-10 has been shown to down-regulate numerous macrophage functions, including microbicidal activity against intracellular bacteria and parasites. IL-10 also inhibits interferon-gamma (IFN-gamma) production and antigen-specific proliferation of Th1 cells mediating immunologic resistance to mycobacterial infection. In contrast, TNF-alpha activates macrophages and may augment their mycobacterial activity. In this study, peripheral blood mononuclear cells (PBMC) or blood monocytes obtained from healthy tuberculin reactors were stimulated in vitro with heat-killed Mycobacterium tuberculosis or heat-killed M. avium-intracellulare complex (MAC) to produce IL-10 and TNF-alpha. We studied a total of 26 clinical isolates of M. tuberculosis and 28 isolates of MAC. MAC-stimulated PBMC and monocytes released significantly larger amounts of IL-10 than those cells stimulated with M. tuberculosis. However, there was no difference in induction of TNF-alpha production between MAC and M. tuberculosis. When TNF-activity was neutralized by the addition of anti-TNF-alpha mAb in culture, MAC still induced more IL-10 secretion than did M. tuberculosis. These findings suggest that increased production of IL-10 by MAC-stimulated monocytes may play a role in the intractable disease caused by these organisms. PMID- 9745228 TI - [Clinical efficacy of fosfomycin in combination with sulbactam/cefoperazone in the treatment of severe infections complicated to blood dyscrasia. Working Group of Kanto Combination Therapy for FOM + SBT/CPZ]. AB - In the treatment of severe infections complicated to blood dyscrasia, the efficacy and usefulness of fosfomycin (FOM) in combination with sulbactam (SBT)/cefoperazone (CPZ) were compared between patients receiving FOM in the first followed by SBT/CPZ (Group A) and those receiving both drugs simultaneously (Group B). The following results were obtained. 1. The efficacy rate was 56.3% for Group A and 47.9% for Group B, with no significant difference. 2. The efficacy for patients suspected of the presence of septicemia, the efficacy rate was 57.9% for Group A and 54.3% for Group B, with no significant difference. 3. As for underlying disease, patients with acute myelogenous leukemia were most prevailing. In these patients, the efficacy rate was 57.1% for Group A and 27.3% for Group B, with no statistically significant difference. However, the efficacy rate tended to be higher in Group A. 4. The administration of antibiotics was effective to restore the neutrophil count to 501/microliters or higher in 77.8% and 45.5% of the cases for Groups A and B, respectively, with significantly higher efficacy for Group A. 5. In the safety evaluation a total of 115 cases were included. Side effects and laboratory abnormalities were seen in 3 cases each, but none of them were serious in degree. From these results, it was confirmed that the combination therapy consisting of administration of FOM followed by SBT/CPZ with some interval is effective for severe infections complicated to blood dyscrasia. PMID- 9745229 TI - [A case of encephalitis due to Mycoplasma pneumoniae: detection of specific DNA from cerebrospinal fluid and elevation of interleukin-6]. AB - A 9-year-old female was admitted to our hospital due to a generalized seizure and consciousness disturbance. The patient had a fever and rash four days before admission, but she had no respiratory symptoms. The seizure and consciousness disturbance was prolonged and intractable. We diagnosed the patient as having encephalitis because of the increase in the cell count in the cerebrospinal fluid (CSF) and a diffuse slow EEG wave. The computed tomography of the head was normal. The causative agent was identified as Mycoplasma pneumoniae because of the increase of antibodies, and the detection of a specific DNA with a polymerase chain reaction. The interleukin (IL)-6 level of CSF was high (384 pg/ml). In spite of intensive treatment she had severe neurological sequelae. The invasion of Mycoplasma pneumoniae to the central nervous system appeared to have a role in the development of encephalitis in the patient. We speculated that there is a possible relationship between the IL-6 levels of CSF and clinical severity of encephalitis. PMID- 9745230 TI - [A fatal case of streptococcal toxic shock-like syndrome probably caused by acupuncture]. AB - A 41-year-old male received acupuncture in the right shoulder for the sake of arthralgia. Three days after acupuncture he was admitted due to severe epigastralgia. Erythematous change and swelling were observed around the right shoulder. A study by magnetic resonance showed an increased signal intensity in a portion of the right subscapular muscle. Four hours after admission he became hypotensive. The erythematous and necrotic change in the right shoulder skin rapidly spread. Excisional debridement in the right lateral chest wall was immediately done. However, the patient died one day after admission despite administration of a high-dose ampicillin and other supportive therapies. Bacteriological and histological examinations confirmed severe streptococcal myositis. This is a case report of toxic shock-like syndrome probably caused by acupuncture. PMID- 9745231 TI - Cognitive brain activity in Alzheimer's disease: electrophysiological response during picture semantic categorization. AB - Semantic memory deterioration is a major component of the cognitive decline seen in patients with dementia of the Alzheimer's type (DAT); however, the exact nature of this deficit remains unclear. Some research data support a procedural deficit where there is an inability to access or retrieve the contents of semantic memory, while other data point to a degraded semantic store where the actual content of semantic memory is degraded. Additional information about semantic processing in DAT can be obtained through the use of an event-related potential (ERP) component known as N400. In the present study, ERPs were recorded from 10 young control participants, 10 elderly control participants, and 10 DAT patients in a picture-semantic matching task. Stimuli were presented sequentially as prime-target pairs, with one-half of the targets matching the primes via semantic relationships (e.g., piano-violin) and the other half mismatching the prime (e.g., helmet-violin). The task was to discriminate between semantically related and unrelated pairs of pictures. In the young and elderly control groups, ERPs generated a larger N400 for unrelated than related target pictures, with a maximum amplitude around 380 ms in the young group and around 480 ms in the elderly group. The amplitude of the N400 was significantly reduced in the DAT patients. However, a separate analysis of congruent and incongruent ERPs trials revealed significant differences only with the incongruent trials. The amplitude of incongruent recordings was larger for the elderly control group than for the DAT patients, while the amplitude for congruent recordings was similar in both groups. These findings are consistent with the neuropathological evidence that Alzheimer's disease is a neocortical disconnection syndrome in which there is a loss of structural and functional integrity of long corticocortical tracts. The semantic activation created by the context is not used efficiently in processing stimuli, which affects access to specific concepts and gradually leads to a breakdown in the structure and organization of semantic memory. PMID- 9745232 TI - Cognitive and motor slowing in Alzheimer's disease and geriatric depression. AB - While response slowing on psychological tasks is a symptom of both depression and Alzheimer's disease (AD), the underlying mechanisms may be quite different: a slowing of cognitive processing in AD and a motor retardation in depression. This hypothesis was tested by examining the rate at which participants performed a simple cognitive operation: subvocal pronunciation. Participants were shown words of between one and three syllables and were asked to decide whether each word ended in a particular sound. This task required participants to transform the written word into its phonological representation, an operation thought to involve subvocal pronunciation. Decision time rose linearly with the number of syllables in all three subject groups. The linear function of the AD patients had a significantly greater slope, indicating a slower rate of subvocal pronunciation, whereas the slope was the same for the normal old and depressed. Both the depressed and AD patients had a higher intercept than the normal old, suggesting a sensorimotor slowing. After treatment, the intercept of the linear function for depressed patients fell, but there was no change in the slope. Thus, this study suggests that AD produces a slowing in both cognitive and motor processes, whereas depression results solely in a motor retardation. PMID- 9745233 TI - Object decision priming in Alzheimer's disease. AB - Priming for line drawings of real and nonreal objects was examined in an object decision task for 16 patients with Alzheimer's disease (AD) and 16 normal elderly control (NC) participants. In two study phases, participants decided if objects were real or nonreal. In an implicit test phase, real/nonreal decisions were made for studied and unstudied objects, and priming was measured as the difference in decision speed or accuracy between studied and unstudied objects. In an explicit test phase, yes/no recognition was measured for real and nonreal objects. AD patients had impaired explicit memory for real and nonreal objects and intact repetition priming for real objects. By the latency measure, both AD and NC groups showed priming for nonreal objects but in opposite ways. Classification decisions about studied relative to nonstudied nonreal objects were slower for the AD patients, whereas such decisions were faster for the NC participants. Classification decisions of both groups were less accurate for repeated nonreal objects. These results support the claim that AD patients with mild cognitive impairment show normal perceptual priming. The AD inhibition for studied nonreal objects is discussed in terms of the decision conflict that occurs when recollection of source is not available to counter the influence of familiarity. PMID- 9745234 TI - Personality and social competency following unilateral stroke. AB - Neuropsychological research indicates that the left hemisphere plays a dominant role in verbal production and processing, while the right hemisphere plays a dominant role in nonverbal production and processing. This study sought to examine the effects of such differential hemispheric specialization on personality and social competency. Ten left hemisphere damaged (LHD) stroke patients, 11 right hemisphere damaged (RHD) stroke patients, and 7 neurologically normal (NHD) patients were videotaped while engaging in social interaction with their spouse and an interviewer. Segments of the interactions were independently coded by two observers. Patients and spouses were rated with respect to their level of social competency and the extent to which they were characterized by 10 personality adjectives (e.g., outgoing, warm). Ratings for the personality items were summed to create an aggregate score. Analysis of these scores revealed both LHD and RHD patients to have lower (i.e., more negative) mean scores than NHD patients, suggesting that stroke patients as a whole were seen as socially impaired. Analysis of the socially competent item revealed particular LHD deficits; LHD patients were seen as less socially competent than both RHD and NHD patients. Spouses of LHD, RHD, and NHD patients, in contrast, did not differ in observer-rated social behavior. PMID- 9745235 TI - Attention and verbal learning in patients with chronic fatigue syndrome. AB - Former neuropsychological studies with Chronic Fatigue Syndrome (CFS) patients evaluated a broad range of cognitive functions. Several, but not all, reported subtle attentional and memory impairments suggesting possible mild cerebral involvement. In this study, a battery of attentional tests and a verbal memory task were administered to 20 CFS patients and 22 healthy controls (HC) in order to clarify the specific nature of attention and memory impairment in these patients. The results provide evidence for attentional dysfunction in patients with CFS as compared to HC. CFS patients performed more poorly on a span test measuring attentional capacity and working memory. Speeded attentional tasks with a more complex element of memory scanning and divided attention seem to be a sensitive measure of reduced attentional capacity in these patients. Focused attention, defined as the ability to attend to a single stimulus while ignoring irrelevant stimuli, appears not to be impaired. CFS patients were poorer on recall of verbal information across learning trials, and poor performance on delayed recall may be due to poor initial learning and not only to a retrieval failure. PMID- 9745236 TI - When does Huntington's disease begin? AB - Recent studies have detected basal ganglia atrophy in clinically asymptomatic persons with the genetic mutation that causes Huntington's disease (HD). Whether reductions in caudate and putamen volume on MRI scans are associated with changes in cognitive and neurologic functioning was examined in 13 healthy adults with the IT-15 mutation. Reduced striatal volume was found to correlate with greater neurologic (largely motor) impairment, slower mental processing speed, and poorer verbal learning, although none of the participants met even liberal criteria for clinical diagnosis of HD. These correlations are strikingly similar to those observed in symptomatic HD patients, possibly reflecting the earliest manifestations of disease. PMID- 9745238 TI - Cerebellar contribution to linguistic processing efficiency revealed by focal damage. AB - The cerebellum's role in cognitive skills was examined in a child (L.C.) with focal injury to the left cerebellum. Initial symptoms included aphasia and dysarthria. At 3 and 9 months post-injury, clinical neuropsychological tests revealed persistent psychomotor slowing as well as deficits in executive functions. Further cognitive testing at 13 and 16 months post-injury demonstrated that L.C. processed information from both the linguistic and nonlinguistic domains more slowly than age-, grade- and sex-matched controls. Notably, her linguistic processing was more than twice as slow as that of her peers, whereas her nonlinguistic processing was only approximately 20% slower. Within each domain the degree of cognitive slowing was approximately the same across diverse tasks. These results are consistent with the hypothesis of a cerebellar contribution to cognitive processing, particularly the processing of linguistic information. PMID- 9745237 TI - A study of performance on tests from the CANTAB battery sensitive to frontal lobe dysfunction in a large sample of normal volunteers: implications for theories of executive functioning and cognitive aging. Cambridge Neuropsychological Test Automated Battery. AB - Several tests from the CANTAB neuropsychological test battery previously shown to be sensitive to frontal lobe dysfunction were administered to a large group of normal volunteers (N = 341) ranging in age from 21 to 79 years. The main tests included a computerized form of the Tower of London test of planning, a self ordered spatial working memory task, and a test of attentional set formation and shifting. A computerized form of the Corsi spatial span task was also given. Age related graded declines in performance were seen, sometimes in a discontinuous manner, especially for the attentional set shifting task (at the extradimensional shift stage). Patterns of deficits reminiscent of frontal lobe or basal ganglia damage were observed in the oldest age group (74-79). However, overall the data were only partially consistent with the hypothesis that frontal lobe functions are the most sensitive to effects of aging. Factor analyses showed that performance in the executive tests was not simply related to a measure of fluid intelligence, and their performance had a factor loading structure distinct from that for the CANTAB tests of visual memory and learning previously administered to the same sample. Finally, only limited support was found for the hypothesis that cognitive aging depends on slowed information processing. PMID- 9745239 TI - Differential lateralization of memory discrimination and response bias in temporal lobe epilepsy patients. AB - Recognition memory for words and designs was assessed in epilepsy patients who underwent unilateral anterior temporal lobectomy. Memory was assessed during the intracarotid amobarbital test (IAT) performed prior to surgery and also following surgery. Memory discrimination and response bias lateralized differently. Memory discrimination, or memory accuracy, lateralized as a function of the type of material used in memory testing. Left temporal lobe lesions resulted in more impaired discrimination of verbal materials; right temporal lobe lesions resulted in more impaired discrimination of visuospatial materials. Response bias, the decision rule adopted in situations of uncertainty, was more liberal following left temporal lobe lesions for both verbal and visuospatial materials. Findings suggest that the two cerebral hemispheres are differentially specialized for encoding different types of information in long term memory, and that this impacts on decision strategies in situations of memory uncertainty. PMID- 9745240 TI - The rate of progression of Alzheimer's disease in the later stages: evidence from the Severe Impairment Battery. AB - This study describes changes in cognitive function in patients who are typically excluded from longitudinal studies due to their inability to complete most standardized batteries. Thirty-three patients with moderate to severe Alzheimer's disease were assessed by means of the Severe Impairment Battery (SIB), Mini Mental State Examination (MMSE), Clinical Dementia Rating (CDR), and measures of functional status. Follow-up evaluations were conducted, and annual rates of change were calculated. Patients with severe cognitive impairment demonstrated rates of progression similar to patients who were moderately impaired at initial evaluation. Rates of decline on the SIB were unrelated to baseline cognitive function, and annual rate of decline in Year 1 was not correlated with disease progression in the subsequent year. This study demonstrates that, with appropriate assessments, severely impaired patients can exhibit a range of performance and rates of decline that are comparable to those of moderately impaired patients. PMID- 9745241 TI - The central executive does not exist. AB - In this article I critically review the concept of the central executive. I argue that the experimental evidence for a central executive lacks rigor to the point where it is an unfalsifiable construct. I examine the neuropsychological and neuroradiological evidence and demonstrate that there is no localization evidence for a central executive. What emerges instead is a pattern of extensive heterogeneity with different executive tasks associated with different neural substrates. In sum it is argued that the idea of a central executive should be abandoned, and, from a neuropsychological perspective, tests that purport to measure executive function should do so in a qualitative way rather than assume that a range of tests load on a unitary dimension of performance. PMID- 9745242 TI - The central executive: a concept and some misconceptions. AB - Parkin's criticisms of the central executive are based on a series of misconceptions. The central executive is not an organ that might or might not exist, but a scientific concept. Part of its function is to separate the analysis of executive processes from the question of their anatomical location. Like other components of working memory, it is fractionable into subsystems. How the subsystems interrelate and how they map onto the anatomical substrate are empirical questions under active current investigation. PMID- 9745243 TI - Mitigating mental retardation in capital cases: finding the "invisible" defendant. PMID- 9745244 TI - Ultrastructure of obstructive tissue in malfunctioning ventricular catheters without infection. AB - The obstructive tissue in eight malfunctioning ventricular catheters without infection was studied using scanning and transmission electron microscopy. Shunt obstruction was due to debris from ventricular structures such as the choroid plexus and ependymal tissue. There was a preponderance of collagen fibers, and many fibroblasts were present within these tissues. The cytoplasm of the fibroblasts contained extended endoplasmic reticulum. The tissues filled the lumen of the catheters in radially arranged layers. Peeling the silicone was seen on the surface of the ventricular catheters but was not present in normal silicone catheters. Finger-like microvilli were observed on the free surface of the tissue. Many vessels were seen in the transverse section of the tissue. Activated fibroblasts and vascularization may be important in tissue growth in ventricular catheters. PMID- 9745245 TI - Multiple scalp aneurysms caused by atypical temporal arteritis--case report. AB - A 55-year-old male suffered sudden onset of dysarthria and mild left hemiparesis due to a right intracerebral small hemorrhage. On admission, six subcutaneous elastic hard lumps were found on the scalp with painless and regular pulsation. The lumps were located along the course of the bilateral superficial temporal arteries (5 locations) and the occipital artery. The patient did not have symptoms of headache or blurred vision associated with temporal arteritis. The largest lump was removed for cosmetic reasons and definitive diagnosis. Histological examination demonstrated many infiltrating inflammatory cells along the entire vascular wall but without giant or fibrinoid necrosis. These multiple scalp aneurysms were probably caused by atypical temporal arteritis. PMID- 9745246 TI - Fenestration of the proximal anterior cerebral artery (A1) with aneurysm manifesting as subarachnoid hemorrhage--case report. AB - A 50-year-old female presented with rare fenestration of the proximal anterior cerebral artery (A1 segment). This rare congenital anomaly was associated with an aneurysm at the proximal end of A1 fenestration causing subarachnoid hemorrhage, and aplasia of the contralateral A1 segment. Surgical treatment to clip the aneurysm resolved her symptoms. Hemodynamic stress at the arterial fenestration probably caused the aneurysm, possible induced by aplasia of the contralateral A1 segment. PMID- 9745247 TI - Temporal lobe epilepsy associated with cerebral venous angioma--case report. AB - A 21-year-old male presented with temporal lobe epilepsy associated with a venous angioma in the ipsilateral frontal lobe, presenting as intractable complex partial seizures. Neuroimaging showed a cerebral venous angioma in the right dorsolateral and opercular frontal lobe, and atrophy of the right hippocampus. As the ictal electroencephalogram (EEG) obtained with subdural electrodes indicated spike discharges initiating from the right mesial temporal lobe, temporal lobectomy was performed. The patient was seizure-free after the operation. Patients with epilepsy who have a cerebral venous angioma require precise analysis of the seizure pattern and an ictal EEG because of cerebral venous angioma may be associated with an another epileptogenic lesion which is surgically treatable. PMID- 9745248 TI - Embolic stroke in the territory of a cerebellar arteriovenous malformation--case report. AB - A 63-year-old male presented with an embolic stroke in the right medial inferior cerebellum. Angiography at 2 and 8 months after the stroke revealed arteriovenous shunts from the vermian branch of the right posterior inferior cerebellar artery of the right inferior vermian vein. The shunts mimicked the arteriovenous shunts of post-recanalization in acute ischemic stroke, but were finally diagnosed as a pre-existing congenital arteriovenous malformation based on their persistence. Arteriovenous shunts that persists more than 2 weeks after ictus should be differentiated from the post-recanalization arteriovenous shunts of ischemic stroke, as the different etiology may affect the ultimate prognosis and course of treatment. PMID- 9745249 TI - Hyperperfusion syndrome after extracranial-intracranial bypass in a patient with moyamoya disease--case report. AB - A 47-year-old female developed hyperperfusion syndrome after superficial temporal artery-middle cerebral artery (STA-MCA) bypass for moyamoya disease. She presented with right hemiparesis and motor aphasia due to left cerebral infarction. She underwent left STA-MCA bypass. One day after surgery, she manifested neurological deterioration. Magnetic resonance (MR) imaging 4 days after the operation indicated regional edema in the territory supplied by the bypass, and single photon emission computed tomography 17 days after the operation demonstrated hyperperfusion in that area. Symptoms improved within 1 week after surgery, and MR imaging showed disappearance of edema and return to the preoperative appearance. Such events are rare, but hyperperfusion syndrome may occur after STA-MCA bypass for moyamoya disease. PMID- 9745250 TI - Ossification of transverse ligament of the atlas associated with atlanto-axial dislocation--case report. AB - A 68-year-old male presented with progressive quadriparesis. Twelve years previously he had undergone anterior decompression with bone grafting for cervical ossification of the posterior longitudinal ligament associated with spinal trauma. Radiological examination showed ossification of the transverse ligament of the atlas (TLA) and severe stenosis of the upper cervical canal. Anterior dislocation of the atlas was also present, but the occiput/atlas/axis unit was perfectly stable due to the prior anterior fusion. Suboccipital decompression and laminectomy of the atlas were performed, and his symptoms improved. Based on the atlanto-axial dislocation, the TLA might have been damaged at the time of the primary trauma and became the trigger for the ossification of the TLA, showing that marked ossification of a vertebral ligament can occur after injury. PMID- 9745251 TI - Acute cauda equina syndrome secondary to lumbar disc herniation mimicking pure conus medullaris syndrome--case report. AB - A 49-year-old male presented with a rare case of acute cauda equina syndrome secondary to a sequestrated lumbar disc mimicking pure conus medullaris syndrome. He consulted the emergency room of Otsu Municipal Hospital because saddle anesthesia and complete urinary retention which had started 2 days before admission. Additionally, he complained of constipation and impotence. Abnormality of the deep tendon reflexes, sciatica, and leg weakness were not organized. Emergent myelography and subsequent computed tomography revealed a sequestrated disc in the sacrum compressing the lower cauda equina, but not the conus medullaris. Emergent surgical decompression was performed about 60 hours after the onset. The patient almost fully recovered from sphincter dysfunction, impotence, and saddle anesthesia 3 months after the operation. When a syndrome like conus medullaris compression is encountered, the lower cauda equina should be examined as well as the conus level. Such cases require urgent diagnosis and treatment. PMID- 9745253 TI - Treatment of the survivors from the Nagoya air crash. AB - Six patients were taken to our hospital alive just after the air crash at Nagoya Airport in 1994. On admission, all patients suffered from profound shock which rapidly progressed. Serum albumin and hemoglobin levels, and platelet count decreased on admission or soon after. The four patients who died could not recover from the shock and associated rapid deterioration of neurological signs. However, two patients were successfully treated with massive transfusion of packed red blood cells, fresh frozen plasma, fresh blood, and/or platelet concentrate and survived to discharge. We recommend addition of albumin to the transfusate in such cases. PMID- 9745252 TI - Subarachnoid hemorrhage caused by Aspergillus aneurysm as a complication of transcranial biopsy of an orbital apex lesion--case report. AB - A 62-year-old male complaining of unilateral visual disturbance and pain in the involved eye had a small mass at the right orbital apex which was identified as an Aspergillus granuloma by transcranial biopsy. One month later, the patient became comatose because of fatal subarachnoid hemorrhage due to a newly developed aneurysm. Autopsy showed a ruptured aneurysm on the right internal carotid posterior communicating artery. Histological examination demonstrated prominent Aspergillus invasion of the arterial wall. Aspergillus infection must be taken into consideration in patients with orbital apex syndrome, which may lead to serious cerebrovascular consequences. If sino-orbital lesions are detected by neuroimaging techniques, biopsy using an extradural approach should be performed to obtain a definitive diagnosis. PMID- 9745254 TI - [Diagnostic abilities of high-resolution CT, dynamic CT, and 201T1 SPECT in evaluating of pulmonary masses]. AB - The purpose of this study was to evaluate the diagnostic abilities of high resolution CT (HRCT), dynamic CT (DCT), and T1-201 SPECT (T1) in determinating benignancy and malignancy (b-m) in pulmonary mass lesions. The diagnoses (35 adenocarcinomas, 10 squamous cell carcinomas, 6 other primary lung carcinomas, 8 tuberculomas, 13 other benignancies) were made in 20 patients (pts) by surgery, in 46 pts by biopsy or cytology, and in 6 pts by clinical course. In 72 pts (51 malignancies, 21 benignancies) who underwent DCT, increased attenuation of lesions at 90 seconds after the injection of contrast medium was a discriminative indicator, and the b-m threshold was defined as 22HU and 15HU for lesions of < or = 3cm and 3cm < in maximum diameter. In 56 pts (43 malignancies, 13 benignancies) examined by T1, lesion-to-contralateral normal lung ratios at 15 min (ER) and 3 hr (DR) were calculated, and the retention index (RI) was defined as (DR-ER)/ER 100. The b-m threshold of RI proved to be-6 for lesions of all sizes. In 40 pts (29 malignancies, 11 benignancies) who underwent both DCT and T1, HRCT was read on the basis of morphology by 7 observers (3 experienced, 2 senior, and 2 junior radiologists). Sensitivity and specificity were 88.2% and 71.4%, respectively, for DCT in 72 pts and 83.7% and 84.6% for T1 in 56 pts. A-receiver-operating characteristics (ROC) analysis revealed that only 2 experienced radiologists were superior to DCT and T1 in diagnostic accuracy. Sensitivity and negative predictive value were 100% and 100%, respectively, for the combination of DCT and T1, and 96.6% and 85.7% for the combination of the 2 experienced radiologists. In conclusion, the combination of DCT and T1 has excellent clinical efficacy in assessment b-m in pulmonary mass lesions. PMID- 9745255 TI - [Evaluation of the cystic duct and cysticohepatic junction with MR cholangiography: comparison of various techniques and clinical evaluation with respiratory-triggered three-dimensional multislab technique]. AB - The purpose of this study was to evaluate depiction of the cystic duct and cysticohepatic junction by MR cholangiography (MRC). In 10 volunteers, MR cholangiograms were obtained by breath-hold two-dimensional (2D), respiratory triggered 2D, respiratory-triggered three-dimensional (3D) single slab, and 3D multislab techniques. The images then were compared qualitatively. MRC using the respiratory-triggered 3D multislab techniques was evaluated as better than the other techniques, and was performed in 35 patients. Depiction of the anatomy of the cystic duct and cysticohepatic junction were evaluated. In 8 of 35 patients, MRC images were compared with those obtained by endoscopic retrograde cholangiography (ERC). The cystic duct and cysticohepatic junction were visualized adequately in 93% of volunteers and patients by the respiratory triggered 3D multislab technique. Anatomic variations in the cystic duct and cysticohepatic junction were evaluated. The frequency of anatomic variations was the same as previously reported. The anatomic evaluations obtained by MRC were correlated closely with those obtained by ERC in 8 patients. In conclusion, MRC with the respiratory-triggered 3D multislab techniques is useful in evaluation of the cystic duct and cysticohepatic junction. PMID- 9745256 TI - [Double oblique MR images of the shoulder: comparison with conventional images]. AB - PURPOSE: Because the scapula is not only slanted on transverse sections but also inclines on sagittal section, we now perform shoulder MR imaging using double oblique images (DOI), which are planes perpendicular or parallel to the long axis of the scapula obtained with oblique sagittal scout imaging. The purpose of this study was to evaluate the usefulness of double oblique shoulder MR imaging. MATERIALS AND METHODS: MR images of shoulders with operatively or arthroscopically proven lesions (20 cases) that had been examined on both conventional images (CI) and DOI were retrospectively reviewed. DOI were compared with CI not only in terms of diagnostic performance but also in their ability to identify the details of shoulder anatomy. All MR studies were done with a shoulder coil on a high-field (1.5T) unit. RESULTS: Although the accuracy of DOI in diagnosing shoulder disorders such as rotator cuff tear and labrum injury was not as good as that of CI, DOI were better for identifying or discriminating muscles and tendons of the rotator cuff, labralbicipital junction and anterior band of the inferior gleno-humeral ligament, and for recognizing the correct position of the glenoid labrum. CONCLUSION: MR double oblique imaging of the shoulder provides more detailed information about shoulder anatomy and disorders than conventional imaging. PMID- 9745257 TI - [Differentiation between benign and malignant breast lesions using fat-suppressed dynamic MR imaging]. AB - PURPOSE: To assess the value and problems of fat-suppressed dynamic MR imaging in differentiating between benign and malignant lesions. MATERIALS AND METHODS: In twenty-nine patients who underwent excisional biopsy or surgical resection, fat suppressed dynamic MR imaging was performed with a 0.5 T superconducting magnet. Pre-and postcontrast 3D-spoiled gradient echo sequences were employed with fat suppression. We calculated and evaluated the contrast-to-noise ratio (CNR) and contrast enhancement ratio (CER) at each contrast determination time (CDT), which is the intermediate time in the scan. RESULTS: Time intensity curves of CNR showed no statistically significant difference between cancers and other benign lesions. The difference in CER between malignant and benign disease was highly significant (P = .006) at CDT 45 sec., but there was great overlap in the time intensity curve of CER after CDT 45 sec. CONCLUSION: When we attempt to differentiate malignant from benign breast lesions by dynamic MR imaging, comparison of CNR is impertinent, and we should evaluate the differential diagnosis of cancer versus benign lesions by means of CER at CDT points of about 45 sec. PMID- 9745258 TI - [Efficacy of dynamic susceptibility contrast MRI using echo-planar imaging in differential diagnosis of breast tumors]. AB - It has been shown that T1-weighted dynamic MR imaging is a useful method in differentiating malignant breast tumors from benign lesions. Invasive breast carcinomas enhance more rapidly than benign lesions such as fibroadenomas, papillomas, and proliferative fibrocystic diseases. However, significant overlap in the dynamic profile of benign and malignant lesions may occur, resulting in relatively low specificity, which is an inherent limitation of this technique. The author attempted to improve diagnostic accuracy by utilizing dynamic susceptibility contrast MR imaging (DSC-MRI) with a single-shot echo-planar imaging sequence. Twenty-two patients underwent DSC-MRI using a 1.5-T unit (Magnetom Vision, Siemens). Images were obtained before, during and after the bolus injection of 20 mL of gadopentetate dimeglumine. The signal reduction rate within the first 30 seconds (delta RT2) was calculated by the following equation: delta RT2 = (postcontrast signal intensity-precontrast signal intensity)/precontrast signal intensity. A rapid, strong decrease in signal intensity was observed on the first pass of the contrast material in all cases of carcinoma, whereas no or only a minimal decrease in signal intensity was observed in all but one of the benign lesions. This method seems to be more accurate than T1-weighted dynamic MR imaging in the differentiation benign and malignant breast lesions. Since DSC-MRI can be performed quickly, subsequent conventional T1 weighted imaging can provide additional information about the morphologic features of lesions, to further support the diagnosis. In conclusion, DSC-MRI seems to be a promising method for the accurate preoperative assessment of breast lesions. PMID- 9745259 TI - [Report management system using Web server and search engine]. AB - PURPOSE: To develop a management system of written reports using a Web server and search engine. METHODS: All the reports written through word processing software in the local area network in the radiology department were automatically transferred to one of the PC servers, in which a Web server and search engine were in operation. Previous reports, approximately eighty thousand, were also registered. Using Web browsers and the program developed by hypertext mark-up language and JavaScript, all the reports were readily accessible by keywords, or by any words within reports. RESULTS: Over 100, 000 reports were easily accessible using Web browsers, based on our one-year experience. Maintaining the Web server and search engine, and upgrading the program were easily managed in our daily practice. CONCLUSION: The report management system we developed is flexible, scalable, and easy to maintain. It is an effective way to manage a large volume of radiology reports written through word processing software. PMID- 9745260 TI - [Cone beam CT utilizing a rotational digital radiographic system: early clinical experience on pulmonary and skeletal lesions]. AB - Utilizing a rotational digital radiographic system (SF-VA 100, Hitachi) 289 digital images were obtained during a 360-degree rotation in 4.8 seconds. The images were transferred to a high-speed workstation and post-processed to create volume data in 9 minutes. The multi-planar reconstruction method reconstructed high-quality tomographic images with equal resolution in any direction. We applied this system as a cone beam CT in order to demonstrate pulmonary and spinal lesions. Early clinical experience suggested the usefulness of the method in the evaluation of diseases in organs with high radiographic contrast. PMID- 9745261 TI - [The influence of sleep breathing disorder on growth hormone secretion in children with tonsil hypertrophy]. AB - INTRODUCTION: Recently it has been revealed that Obstructive Sleep Apnea Syndrome has an influence on the endocrine system, especially to the secretion of growth hormone (GH). It was previously reported that secretion of GH has a circadian rhythm along with a correlation with slow wave sleep. Normally in a pediatric patient, after 90 min of sleep the brain wave shows patterns of deep sleep, and a matching peak in GH secretion is observed. We have studied the effect of GH secretion in sleep breathing disorder of pediatric patients by observing the pre- and post-operative difference in GH secretion in children with tonsillar hypertrophy who had tonsillectomies. MATERIAL AND METHODS: Ten pediatric patients who complained of sleep apnea or hypopnea, and were highly suspected of having sleep breathing disorders due to tonsil hypertrophy and underwent surgery were chosen. Pre and post-operative blood somatomedin C and urinary growth hormone levels were measured. In 6 of the 10 patients blood GH levels were measured after overnight collection of blood every 30 min through a catheter placed in the vein. METHOD OF ANALYSIS: Integral blood GH calculated from GH levels of the blood samples taken every 30 min was used to express the efficacy of GH secretion during sleep, and was compared with blood somatomedin C and urinary GH levels. Pre- and post-operative somatomedin C and urinary GH measurements were compared. RESULT: 1) In 6 of the 10 patients, there was a positive correlation with integral GH and urinary GH, and no correlation with integral GH or somatomedin C. 2) Post-operatively, 7 of the 10 patients had increased urinary GH levels, 1 showed no change and 2 had decreased levels. There was a significant increase in the mean urinary GH levels from 18.6 +/- 16.1 pg/ml pre-operatively to 35.8 +/- 19.6 pg/ml post-operatively (P < 0.05). 3) Eight of the 10 patients had a post operative increase in somatomedin C levels, 1 showed no change and the other patient had a decreased level. There was a significant increase in mean somatomedin C levels from 175.9 +/- 124.8 ng/ml pre-operatively to 226.6 +/- 134.3 ng/ml post-operatively (P < 0.01). CASE: An 8-year-old-boy was diagnosed as having habitual tonsillitis and suspected sleep breathing disorder, and was admitted to our hospital for a tonsillectomy. Apnea and hypopnea were observed during sleep in the hospital. After receiving his parents' consent, GH secretion was evaluated before and after the tonsillectomy. Integrated GH measurements, somatomedin C and urinary GH increased significantly after the operation. CONCLUSION: In this study, the improvement of sleep breathing disorder by surgery was associated with a positive effect to the GH secretion. GH secretion is important for growth of a child. Wen the cause of sleep breathing disorder is evident in the upper airway system, active treatment and cure by surgery is advised. Measurement of urinary GH in the morning is useful and simple for evaluating the ability to secrete GH. PMID- 9745262 TI - [Speech audiometry using the American Word Lists for Japanese subjects with normal hearing]. AB - The characteristics of listening to English words for Japanese people were studied in ten normal hearing subjects who had taken English classes to the level of college graduates and had opportunities to learn English continuously. Following pure tone audiometry, speech audiometry was performed using the Central Institute for the Deaf (CID) W-1 and W-22 word lists for English and the 67-S word lists for Japanese. The speech reception thresholds (SRTs) for the CID W-1 lists were significantly higher than average pure tone threshold (PTT), although the SRTs for the 67-S lists were equal to the average PTT. The difference in average SRT between the CID W-1 lists and the 67-S lists was about 15dB, which is statistically significant. The speech discrimination rate for the CID W-22 lists ranged from 78 to 100 percent with an average of 89.5 percent, while all subjects achieved the discrimination rate of 100 percent for the 67-S lists. Analysis with transient matrices of the perceived words demonstrated that the articulation rates were below 90 percent for the consonants /m/, /n/, /p/ and /delta/ . The observed variation in the speech discrimination score and the pattern of confusion among the subjects was assumed to be much more pronounced in noisy conditions. PMID- 9745263 TI - [An experimental study of tumor cell infiltration into temporal bones--route into the inner ear from the arachnoid space]. AB - In humans, metastatic tumors which invaded the temporal bones have been studied in regard to the relationship between histopathologic findings and clinical symptoms. On the other hand, there is no experimental study using an animal model for tumor infiltration into the temporal bone. This study was designed to establish such an animal model and examine the temporal bones histopathologically. Rat thymic lymphoma cell (FTL-A2) were inoculated into the cisterna magna of Wistar rats. The animals were decapitated under deep anesthesia with pentobarbital sodium from the 1st to 8th day after inoculation. Their heads were fixed with Heiden-hain SuSa solution, decalcified, dehydrated, embedded in celloidin, and sectioned horizontally at a thickness of 25 microns. These were stained with hematoxylin and eosin, and histologically examined by light microscopy. Inoculated tumor cells showed active viability in the arachnoid space at a rate of 98%. Two major routes of tumor cell infiltration into the inner ear were found: the cochlear aqueduct and the internal auditory canal. At the early stage after the inoculation, tumor cells infiltrated the scala tympani through the cochlear aqueduct. Invading the fiber of the cochlear nerve, tumor cells infiltrated Rosenthal's canal via the tractus spiralis foraminosus, and passed through Rosenthal's canal and the osseous spiral lamina into the scala tymani. However, tumor cells did not infiltrate the organ of Corti through the habenula perporata. Tractus spiralis foraminosus and habenula perforata functioned as a barrier against tumor infiltration. In a few cases, tumor cells infiltrated over the macula cribrosa into the subepithelial space of the utricule and saccule. The macula cribrosa functioned as a barrier. PMID- 9745264 TI - [Neck Dissection for salivary gland carcinoma]. AB - Between 1986 and 1997, 18 patients with high-grade salivary gland malignancies were treated at our institution. Histologically, 7 of the 18 malignant tumors were adenoid cystic carcinomas. 4 were carcinoma in pleomorphic adenoma, 3 were undifferentiated carcinomas and 4 were others. For treatment of the neck, 11 patients underwent neck dissection, 1 received supraomohyoid neck dissection (SOMH) and 7 received no neck treatment. In our study, there was no difference according to local-regional control as to whether to use neck dissection or not. The result do not suggest that prophylactic total neck dissection for salivary gland carcinoma show an impressive degree of improvement in local-regional control. Total neck dissection should be performed when the neck is clinically positive for a tumor. Based on the above findings, we concluded that SOMH is feasible for submandibullar gland cancer without positive lymph nodes. PMID- 9745265 TI - [Decongesting effect of tramazoline on nasal airway patency and nasal symptoms as evaluated by acoustic rhinometry: an objective study in 30 allergic and six non allergic subjects]. AB - In this prospective clinical study, we investigated the effect of the nasal decongestant tramazoline on nasal airway patency in 30 subjects with perennial nasal allergy to housedust mites and in six non-allergic volunteers who served as controls. The allergic subjects were further subdivided into groups, the severe group and the moderate group, according to the level of specific IgE. We used acoustic rhinometry as an objective method, and the nasal cavity volume (NCV) and the minimum cross-sectional area (MCA) were measured before and 10 minutes after nasal spraying. Before mucosal decongestion, the average NCV in the severe group was 5.94 cm3, which was significantly lower than those in the other two groups: 7 cm3 in the control group and 8.13 cm3 in the moderate group, respectively (P = 0.02). A more significant difference was found when the first 3-cm area of the NCV was evaluated (P = 0.009). Pharmacologic decongestion caused an increase in nasal patency by 54% from the baseline NCV values in the control group, 38% in the moderate group and 48% in the severe group. There was no significant difference among the three groups. Changes in nasal symptoms were also determined by questionnaire. The degree of improvement in subjective nasal congestion was more pronounced in the allergic groups than in the control group. PMID- 9745266 TI - [Study of the function of nerve growth factor in the olfactory tract of the mouse]. AB - The aim of this study was to examine the function of nerve growth factor (BGF) in the olfactory tract of mice. Using the mice which had received unilateral olfactory bulbectomy and in which antibodies to NGF had been continuously infused with into the contralateral olfactory blub, three kinds of analysis were performed: histological analysis of the olfactory epithelium by HE staining, immunohistochemical analysis of the olfactory epithelium using polyclonal antibodies to trk which forms the NGF receptor, and olfactory-mediated behavioral analysis with cycloheximide. These animals had been sacrificed at day 1, 3, 7, 14, 21 or 28. Several findings were obtained as a result of the above analysis. Degeneration of the olfactory epithelium and trk expression by the olfactory cells were observed on day 7, and the olfactory epithelium was incompletely regenerated on day 28. However, trk expression by the olfactory cell was still recognized and the olfactory function was not restored by day 28. These examinations suggest that NGF produced in the olfactory bulb was transported retrogradely to olfactory cells through the olfactory nerves, and was associated with sustaining the existence of those cells and with regenerating the olfactory tract after injury. PMID- 9745267 TI - [Long-term prognosis for tonsillectomy patients with IgA nephropathy]. AB - A retrospective study of 85 patients with IgA nephropathy was undertaken to determine the long-term effect of tonsillectomy. Forty-three patients (24 males and 19 females) had received tonsillectomies (Group A) and 42 patients (17 males and 25 females) had not (Group B). These patients had been followed up for more than 5 years after renal biopsy. The average age at the initial renal biopsy was 25.72 years in Group A, and 33.16 years old in Group B. The average period of renal biopsy to tonsillectomy in Group A was 10.47 months. The average follow-up period was 8 years and 9 months in both groups. At the beginning of treatment, the two groups were well matched in terms of creatinine clearance, urinalysis, and blood pressure. Six patients in Group A and eight patients in Group B were treated with steroids. The glomerular injury detected at the renal biopsy was more extensive in Group A than in Group B. Renal function in the two groups was compared. The clinical remission rate in Group A was significantly higher than in group B (P<0.05). The stable renal function rate in Group A was significantly higher than in Group B (P<0.05). The renal survival rate was 97.7% in Group A and 83.3% in Group B, but there was no significant difference between the two groups. Histologically, the rate of remission of the minor lesion in Group A was significantly higher than in Group B (P < 0.05). Our results showed that tonsillectomy for IgA nephropathy was clinically of great value. PMID- 9745269 TI - [Current states and future prospects of kidney failure]. PMID- 9745268 TI - [Identification of penicillin-resistant Streptococcus pneumoniae in nasopharynx of patient with acute otitis media by PCR]. AB - Streptococcus Pneumoniae is a leading cause of acute otitis media (AOM). For most AOM caused by S. pneumoniae, penicillin is the antibiotic of choice. However, there are some recent reports of clinical resistance to penicillin by S. pneumoniae. The sequences of penicillin binding protein, pbpla, pbp2b and pbp2x, genes of penicillin-resistant S. pneumoniae (PRSP) were more highly divergent than those of penicillin-succeptible S. pneumoniae (PSSP). The polymerase chain reaction (PCR) can easily determine whether an S. pheumoniae isolate is susceptible or resistant to penicillin by amplifying the target gene by using a combination of primers. In this study, clinical isolates (n = 12) were obtained from the nasopharynx of patients with AOM. PCR was used to confirm the identification of an isolate as S. pneumoniae by amplifying the autolysin gene and to detect three PBP genes by amplifying parts of pbp1a, pbp2x and pbp2b. The resistance of S. pneumoniae to penicillin and other beta-lactams has been shown to be associated with mosaic mutations in the pbp1a, pbp2b and pbp2x genes. These findings suggest that rapid identification of PSSP and PISP/PRSP by PCR is possible and very useful for proper treatment of acute otitis media. PMID- 9745270 TI - [Diagnosis and symptoms of kidney failure]. PMID- 9745271 TI - [Severity and pathophysiology of kidney failure]. PMID- 9745272 TI - [Drug-induced kidney diseases]. PMID- 9745273 TI - [Etiology and therapy of acute progressive glomerulonephritis]. PMID- 9745274 TI - [Etiology and therapy of multiple organ failure]. PMID- 9745275 TI - [Etiology and therapy of chronic primary glomerulonephritis]. PMID- 9745276 TI - [Etiology and therapy of secondary chronic glomerulonephritis]. PMID- 9745277 TI - [Etiology and therapy of diabetic nephropathy]. PMID- 9745278 TI - [Etiology and therapy of kidney disease due to hypertension]. PMID- 9745279 TI - [Etiology and therapy of polycystic kidney]. PMID- 9745281 TI - [Long-term hemodialysis for advanced-stage chronic kidney failure and their complications]. PMID- 9745282 TI - [Acute blood purification]. PMID- 9745280 TI - [Etiology and therapy of chronic pyelonephritis]. PMID- 9745283 TI - [Home hemodialysis for therapy of late-stage kidney failure]. PMID- 9745284 TI - [Continuous ambulatory peritoneal dialysis for therapy of late-stage kidney failure]. PMID- 9745286 TI - [Problems in treatment of kidney failure (discussion)]. PMID- 9745285 TI - [Kidney transplantation for therapy of late-stage kidney failure]. PMID- 9745287 TI - [Case of 37 year-old female with encephalomyelitis due to mycoplasma pneumoniae]. PMID- 9745289 TI - [Case of multiple cranial nerve palsy following herpes simplex labialis]. PMID- 9745288 TI - [Case of osteomalacia due to intravenous saccharated ferric oxide]. PMID- 9745290 TI - [Case of malignant ameloblastoma of multiple lung metastasis following surgical removal of the primary site 13 years earlier]. PMID- 9745291 TI - [Case of lupus cystitis diagnosed in the early stage]. PMID- 9745292 TI - [Hereditary non-polyposis colorectal cancer]. PMID- 9745293 TI - [Recent studies on reactive arthritis]. PMID- 9745294 TI - [Present status arrhythmia and new therapeutic approach]. PMID- 9745295 TI - [Heart transplantation]. PMID- 9745296 TI - [The role of the age-related genes in gerontology]. PMID- 9745297 TI - [Disorders associated with aging of blood vessels and scavenger receptors]. PMID- 9745298 TI - [Clinical significance of serum lipoprotein(a) in elderly patients with aortic valve sclerosis]. AB - BACKGROUND: Calcified aortic value disease is increasing with explosively in the elderly. Elevated serum lipoprotein(a) (Lp(a) plays an important role in the pathogenesis of atherosclerosis. Thus, we investigated the relationship between aortic valve sclerosis and serum Lp(a) levels in elderly patients. METHODS: Echocardiography was performed in 97 subjects (77 +/- 7 years, 48 males and 49 females), Lp(a), fasting plasma glucose, and blood pressure were measured at the time of the study. Aortic valve sclerosis was assessed using echocardiography. RESULTS: Aortic valve sclerosis was observed in 63 patients (sclerosis group; 24 males and 39 females) and not in 34 subjects (non-sclerosis group; 24 males and 10 females). Univariable analysis revealed that age, Lp(a) level, and the number of females were higher in the sclerosis group than in the non-sclerosis group (age; 78 +/- 7 vs 74 +/- 7 years, p = 0.0090, Lp(a); cholesterol, triglyceride, and fasting blood glucose did not seem to affect aortic valve sclerosis. In all of 9 patients with serum Lp(a) greater than 60mg/dl aortic valve sclerosis was present. In discriminative analysis, gender (female) (lambda = 0.9038, p = 0.0020) and Lp(a) (lambda = 0.8316, p = 0.0053) were related to aortic valve sclerosis. CONCLUSION: Elevated serum Lp(a) was observed in elderly patients with aortic valve sclerosis. PMID- 9745299 TI - [Radiofrequency catheter ablation therapy in elderly patients with supraventricular tachycardia]. AB - 138 patients with Wolf-Parkinson-White (WPW) syndrome (n = 96), atrioventricular nodal reentrant tachycardia (AVNRT; n = 27) and the other supraventricular tachycardia (n = 15), were divided into two groups, a control group (less than 65 years old; n = 108) and an elderly group (more than 66 years old; n = 30). We then estimated the success rate and safety of radiofrequency ablation for supraventricular tachycardia in elderly patients. For WPW syndrome, there were 76 (97%) successes and 9 (13%) recurrences in the control group (n = 78). In the elderly group of WPW patients, the number of successes was 18 (100%) and the number of recurrences one (63%). In 27 patients with AVNRT, the number of successes was 26 (96%) and there were no recurrences. In 15 patients with some other supraventricular tachycardia, there were 11 patients (73%) successes and one recurrence (11%). Major complications consisted of cardiac tamponade in 2 patients, dissecting aneurysm in one patient and cerebral embolism in one patients. All major complications occurred in patients with WPW syndrome. The cause of the complications, except the cerebral embolism was manipulation of the electrical or ablation catheter. Three of four patients with major complications belonged to the control group. It is possible that radiofrequency catheter ablation for supraventricular tachycardia in elderly patients is safe and highly effective. However, it is still invasive therapy. Ablation on a left accessory pathway by the transaortic valve approach especially needs meticulous care. PMID- 9745300 TI - [Validation of interviewer administration of the Short Form 36 Health Survey, and comparisons of health-related quality of life between community-dwelling and institutionalized elderly people]. AB - The Short Form 36 Health Survey (SF-36) is a questionnaire that is widely used to measure health-related quality of life. Because self-administered questionnaires may not be appropriate for seriously ill or elderly people, we administered the SF-36 to institutionalized elderly people by face-to-face interviews, and tested its reliability and validity. We also compared the SF 36 score of those subjects with the scores of community-dwelling elderly people. We studied 117 people aged 65 or over who were living in residential facilities on Sado island and 62 randomly sampled elderly people who were living in the community. The SF-36 scores of the institutionalized subjects had acceptable ceiling and floor effects, and their internal consistency, concurrent validity, and construct validity were high. The only exceptions were the scores of the "vitality" subscale. Adjusted mean scores on four subscales were higher among the institutionalized subjects than among those living in the community: role limitation due to physical condition, role limitation due to emotional condition, social functioning, and bodily pain. The two groups did not differ with regard to scores on the "mental health" scale, the "vitality" scale, or the "general health perception" scale. We conclude that the SF-36 can be useful for measuring health related quality of life among institutionalized elderly people, if it is administered in face-to-face interviews. PMID- 9745301 TI - [Memory function in aging and Parkinson's disease--an event-related potential study]. AB - The N400 of event-related potentials (ERPs) was recorded in 17 healthy young subjects (mean age 24.4 years), 14 healthy old subjects (healthy control subjects, mean age 62.7 years), and 21 patients with nondemented Parkinson's disease (PD, mean age, 63.8 years) as they listened to words or nonwords. Some words were repeated immediately after the initial presentation (2 sec), while others were repeated after 5 intervening words (12 sec) or after 2 to 4 minutes. The subjects were required to respond to occasional nonwords. Rey's auditory verbal learning test(AVLT) was also performed. The mean N400 amplitude was smaller in patients with PD than in either healthy group, but there was no difference between the two healthy groups. In the young subjects, N400 was attenuated for repeated words, and the attenuation was more pronounced for immediate than for delayed repetitions. N400 attenuation in the old subjects was more markedly reduced as the interval between the 1st and 2nd presentation increased than in the young subjects. In PD patients, attenuation was noted only for immediate repetition. Free recall by the old subjects was impaired relative to the young subjects throughout the AVLT trials, and was even more impaired in patients with PD. In addition, the number of free recalls increased less with the number of trials in patients with PD than in the old subjects. These results may indicate that episodic memory declines with advancing age, and declines even more in patients with PD. In addition, the ability to transform information form short term memory to long-term memory appear to be impaired in patients with PD. PMID- 9745302 TI - [Two cases of Churg-Strauss syndrome with impaired prognosis]. AB - We report two cases of Churg-Strauss syndrome with impaired prognosis. Case 1: A 58-year-old woman started to complain of progressive dysesthesia and muscle weakness of the lower extremities on February 6, 1992 and was admitted to our hospital eight days later. The eosinophil count was 1, 170/microliters and chest X-ray on admission revealed bilateral diffuse interstitial shadows. Necrosis of muscle fibers and significant infiltration of eosinophils into vessel walls were histopathologically demonstrated. Case 2: A 69-year-old woman was admitted to our hospital with productive cough and back pain. An expiratory wheeze was audible in all lung fields and bronchial asthma was diagnosed. Laboratory findings showed eosinophilia (8, 550/microliters), an IgE level of 16, 266 IU/ml and no positive data for allergic bronchopulmonary aspergillosis. A biopsy of subcutaneous nodules in the lower extremity revealed significant edema of vessel walls and infiltration eosinophils and lymphocytes. Churg-Strauss syndrome was diagnosed in both cases, and corticosteroid therapy was successful in alleviating eosinophilia. However, symptoms of vasculitis in case 1, including mononeuritis multiplex, did not improve, and cardiomyopathy in case 2 progressively worsened. Particularly in elderly patients, it is very important to make an early diagnosis and to initiate therapy that does not disturb quality of life, even for diseases generally regarded as having a good or fair prognosis. PMID- 9745303 TI - [Renal tubular acidosis type II secondary to gamma-light chain excretion in an elderly patient with multiple myeloma]. AB - A 73-year-old woman was admitted to the geriatric ward of the University of Tokyo Hospital with anemia, osteepeina, and renal dysfunction. Although symptoms typical of multiple myeloma such as punched-out lesions and hyperproteinemia were not found, protein electrophoresis revealed that lambda type Bence-Jones protein was excreted in urine. Multiple myeloma was diagnosed. Furthermore, renal dysfunction was accompanied renal tubular acidosis type II (proximal type). Renal dysfunction in patient with multiple myeloma in usually caused by so-called myeloma casts in the distal tubules, but renal tubular acidosis type II is rarely observed. It is possible that injury of the proximal renal tubular eithelium by Bence-Jones protein resulted in renal tubular acidosis type II in this patient. PMID- 9745304 TI - [A new radiation-free conditioning in bone marrow transplantation and dibromo mannitol therapy in chronic myeloid leukemia]. AB - A new, radiation-free, conditioning protocol, containing the original Hungarian mitobronitol (DBM) (DBM/ cytosine arabinoside/cyclosphosphamide) has been applied to 36 chronic myeloid leukemia (CML) patients followed by bone marrow transplantation (BMT) from HLA identical sibling donors between 1990-1997. In spite of some prognostically disadvantageous factors (half of them were above 40 years, 10 out of 36 patients were in accelerated phase, the disease history was longer than 2 years in average) the overall survival (30/36) and the leukemia free survival rate (26/36) were in accordance with the best international results. Transplantation-related toxicity was remarkably reduced in comparison to bone marrow transplantation performed by total body irradiation/cyclophosphamide (TBI/Cy) or busulphan/cyclophosphamide (Bu/Cy) conditioning protocols. Acute graft versus host disease was present in lower percentage (9/36) and the number of serious cases was only 2/36. Chronic GVH disease, generally known to be associated with antileukemic effect (GVL), occurred in 25 of cases. Early haematological relapse among the 34 patients with functioning graft occurred in 6 patients which rate is slightly higher than reported after TBI/Cy or Bu/Cy conditioning treatment. There was no relapse among patients transplanted within one year post-diagnosis and patients having CML with accelerated phase. The leukemia free post-transplant period was in association with the chronic GVH disease and full chimeric state. PMID- 9745305 TI - [Rhabdomyoma as a first manifestation of childhood tuberous sclerosis]. AB - Two dimensional echocardiography seems to be the best diagnostic tool for diagnosis of cardiac tumors. In our practice using 2 dimensional echocardiography in pediatric patients from 1984 11 cardiac tumors were diagnosed and followed-up. Four of them the cardiac rhabdomyomata (one of them diagnosed in utero) was the first manifestation of tuberous sclerosis. We summarized the follow-up data from our patients with cardiac rhabdomyomata. The detection of cardiac tumor in the fetal or infant period is of outmost importance in the early diagnosis of tuberous sclerosis and suggest careful follow-up and management. To the best of our knowledge no report of tuberous sclerosis based on echocardiographic diagnosis of cardiac rhabdomyomata has been described in our country. PMID- 9745306 TI - [A new method for the management of perforated veins in the lower extremities]. AB - Elimination of incompetent perforating veins is the effective therapeutic method in the treatment of lower leg ulceration and trophic skin disorders associated with chronic venous insufficiency. The subfascial endoscopic ligation is a new surgical method to treat patients with incompetent perforating veins. 19 patients with severe chronic venous insufficiency or protracted venous ulceration of lower leg were treated. Through an incision of the skin the proximal 1/3 of the lower leg-far away from the dermatosclerotic area-an endoscope is inserted after which the perforating veins are ligated by clips under direct vision. The method is recommended due to its reduced invasiveness and the fair results. PMID- 9745307 TI - [Serum and salivary testosterone levels in erectile dysfunction]. AB - In this study 66 male patients with erectile dysfunction were investigated. The authors measured the testosterone levels in serum and in saliva, which latter represent with good accuracy the serum levels of free testosterone. The mean serum total testosterone level was 17.6 nmol/L (confidence intervals: 15.5 and 20.2 nmol/L, normal range: 10-50 nmol/L). The mean salivary free testosterone level was 218.5 pmol/L (confidence intervals: 198.3 and 239.9 pmol/L, normal range: 200-1000 pmol/L). Low salivary (free) testosterone levels were found in 36.4% of patients, while only in 10.6% of patients had low serum testosterone levels (p = 0.01, by binomial test). Although there is a relationship between serum and salivary testosterone levels (r = 0.41, p < 0.001), the patients with low salivary (free) testosterone levels have in major part a normal serum total testosterone level. These data indicate that a considerable proportion of patients with erectile dysfunction have androgen deficiency. The serum total testosterone level is not a sensitive indicator to detection of hypogonadism. The androgen substitution therapy has a beneficial effect on erectile dysfunction in a significant part of patients. The measurement of free testosterone level in saliva may have an important role both in the diagnosis of diseases characterized by androgen deficiency and hyperandrogenic status. PMID- 9745308 TI - [Repeated resection of hepatocellular carcinoma and its recurrences and removal of pulmonary metastases]. AB - A case of a 67 year old male successfully treated local recurrence of previously radically resected hepatocellular carcinoma with distant metastases of the lung is reported. Two years after a complete removal of the primary liver malignoma a local recurrence was resected which was followed by selective intraarterial cytostatic treatment and chemoembolisation. A three-year disease-free interval was achieved, then recurrence in the liver and lung metastases were detected at the regular check-up. All of the malignant focuses were eliminated by parenchyma sparing surgical resections, then a postoperative adjuvant chemotherapy (FAM scheme) has been followed. After eight months the patient remained symptom-free. The long-term survival seems to justify the aggressive approach of this kind of malignancies. PMID- 9745309 TI - [Achievements of academician Professor Laszlo Haranghy, M.D., on the centenary of his birth]. PMID- 9745310 TI - [In memoriam: Ferenc Gaspar and Adam Raffy, medical writers]. PMID- 9745311 TI - Group A beta-hemolytic streptococcal infections. AB - GABHS is the most common bacterial cause of tonsillopharyngitis, but this organism also produces acute otitis media; pneumonia; skin and soft-tissue infections; cardiovascular, musculoskeletal, and lymphatic infections; bacteremia; and meningitis. Most children and adolescents who develop a sore throat do not have GABHS as the cause; their infection is viral in etiology. Other bacterial pathogens produce sore throat infrequently (e.g., Chlamydia pneumoniae and Mycoplasma pneumoniae), and when they do, other concomitant clinical illness is present. Classic streptococcal tonsillopharyngitis has an acute onset; produces concurrent headache, stomach ache, and dysphagia; and upon examination is characterized by intense tonsillopharyngeal erythema, yellow exudate, and tender/enlarged anterior cervical glands. Unfortunately only about 20% to 30% of patients present with classic disease. Physicians overdiagnose streptococcal tonsillopharyngitis by a wide margin, which almost always leads to unnecessary treatment with antibiotics. Accordingly, use of throat cultures and/or rapid GABHS detection tests in the office is strongly advocated. Their use has been shown to be cost-effective and to reduce antibiotic overprescribing substantially. Penicillin currently is recommended by the American Academy of Pediatrics and American Heart Association as first-line therapy for GABHS infections; erythromycin is recommended for those allergic to penicillin. Virtually all patients improve clinically with penicillin and other antibiotics. However, penicillin treatment failures do occur, especially in tonsillopharyngitis in which 5% to 35% of patients do not experience bacteriologic eradication. Penicillin treatment failures are more common among patients who have been treated recently with the drug. Cephalosporins or azithromycin are preferred following penicillin treatment failures in selected patients as first-line therapy, based on a history of penicillin failures or lack of compliance and for impetigo. GABHS remain exquisitely sensitive to penicillin in vitro. There are several explanations for penicillin treatment failures, but the possibility of copathogen co-colonization in vivo has received the most attention. Treatment duration with penicillin should be 10 days to optimize cure in GABHS infections. A 5-day regimen is possible and approved by the United States Food and Drug Administration for cefpodoxime (a cephalosporin) and azithromycin (a macrolide). Prevention of rheumatic fever is the primary objective for antibiotic therapy of GABHS infections, but a reduction in contagion and faster clinical improvement also can be achieved. Development of streptococcal toxic shock syndrome and necrotizing fasciitis ("flesh-eating bacteria") are rising concerns. The portal of entry for these invasive GABHS strains is far more often skin and soft tissue than the tonsillopharynx. PMID- 9745312 TI - Tall stature. PMID- 9745313 TI - Management of status epilepticus in children. AB - Treatment of SE is based on the age of the patient and the possible underlying etiology. Initial treatment should include a benzodiazepine (lorazepam 0.1 mg/kg or diazepam 0.5 mg/kg). Specimens for laboratory tests should be drawn early in the event of a prolonged seizure and geared toward the clinical presentation and age of the patient. If a seizure lasts longer than 10 minutes, phenobarbital 20 mg/kg dose should be administered with strict adherence to the proper rate of administration. Further seizure activity during drug administration can be treated with additional doses of lorazepam or diazepam. If a seizure lasts longer than 10 minutes, a second long-acting anticonvulsant should be administered, followed by induction of general anesthesia. PMID- 9745314 TI - The diagnosis of rheumatic fever. PMID- 9745315 TI - Childhood obesity. PMID- 9745316 TI - Family violence: implications for the pediatrician. PMID- 9745317 TI - Iron deficiency anemia. PMID- 9745319 TI - ["I dreamed that I was dreaming..." Transcript of the 290th session and commentary]. PMID- 9745318 TI - Updates on measles vaccine. PMID- 9745320 TI - [A score in transference. BIP--experiencing the relationship in psychoanalysis]. AB - The (emotional) experience of reference in psychoanalyses, presented here, has been developed by the author in continuation of a method evolved by Gill and Hoffman 1982 in Chicago, which they termed "The Patient's Experience of the Relationship with the Therapist", to describe the course of analytical work on patient resistance against transference. The "Experience of reference in psychoanalyses" is a rating method that links by means of its two manuals quantitative methods (which are empirical in the conventional sense) with qualitative, clinical-hermeneutical research approaches. After reliable coding of categorial data of a tape recorder transcription, a condensed description of the course and a clinical comment are obtained, the graphic representation of which reads like a score transference. PMID- 9745321 TI - ["Repeating" the transference? The central relationship conflict topic of the 290th session--questions, problems, results]. AB - The following text sums up the results of an examination of a therapeutic session in which a psychoanalyst treated a patient suffering from neurotic depression. We employed the CCRT method which was applied at various levels of abstraction and in two steps of evaluation. The standard evaluation procedure allows for both a more thorough evaluation of "self-relationship episodes" and episodes involving other objects. An integration of "tailor-made" formulations allowed us to analyse another set of "relationship-patterns": namely, those which, in contrast to those most often found in conflict themes, contain positive traits. These results were then discussed clinically to examine these in light of the therapeutic conception of depression. In this context, the question arose as to whether the classic CCRT method can be fruitfully applied in long-term analysis and if this notion should be modified in order to examine the analysis of depression more effectively. A first attempt at such a modification was undertaken in the hope it would allow us to evaluate both the relevance of meaning passages contained in the clinical data and the visible processes of change. PMID- 9745322 TI - [Identification of repetitive relationship patterns using the FRAMES METHOD]. AB - Frames is a method to identify Fundamental Repetitive And Maladaptive Emotion Structures in verbatim transcribed psychotherapy sessions. The 5 steps leading to Frames consist of 1. criteria to select sessions or other material apt to identifying Frames; 2. coding of emotions according to Dahls and Stengels (1978) category system; 3. the construction of an Object Map in which each person or object is located where he/she was referred to in the dialogue; 4. the identification of the narrative structure, i.e. the logical plot structure of the stories that are told about these objects; and 5. the construction of Frames as generalised patterns of behaviour. These 5 steps are demonstrated by means of material from a specimen (hour 290) from a psychoanalytical treatment. PMID- 9745323 TI - [How is transference assessed? Evaluation of the 290th session from the clinical perspective]. AB - In modern view, the transference process is both repetition and change. To grasp specific relationship patterns systematically different content-analytical methods are used in the context of singles case studies. FRAMES, BIP (PERT) and CCRT, as demonstrated in the foregoing articles by Holzer, Herold, Deserno et al. place various accents: expressed emotions, interpersonal experience, and the central relational conflict. They do not lead only to a concentrated reconstruction of the respective situational dynamics obtained according to known rules; they also seem to be able to confirm the clinical experience that repetition and change meet in the patient-analyst relationship. PMID- 9745324 TI - [Chaos theory and complexity in psychiatry]. AB - In spite of the continuing development of psychometric and apparative methods, the determination of mind has not yet been successful. This fact is due to the complexity of the nervous system, which can only be described statistically and as an epiphenomenon, just like any other biological system. Fractal geometry presented in the sixties by Mandelbrot provides the conception of fractal dimension, a numerical term allowing a more adequate description of complex systems. In many fields of medical science attempts are being made to apply this knowledge; in psychiatry, however, methodological problems still remain unsolved. For the present the necessity of a new thinking should be emphasized while trying to determine the functions and dysfunction of the mind. PMID- 9745325 TI - Plasmid profile analysis and antibiotic resistance of Salmonella strains from clinical isolates in Cluj-Napoca. AB - Resistance patterns, plasmid profiles and the genetic resistance determinants were investigated in 38 isolates of Salmonella enterica serotype Typhimurium and 19 isolates of Salmonella enterica serovar Enteritidis derived from children hospitalized in two clinics in Cluj-Napoca, during the period of 1995-1997. Incidence of plasmid and antibiotic resistance was very high in Salmonella typhimurium isolates. All strains were resistant to almost all antibiotics tested but susceptible to the third generation cephalosporines and fluoroquinolones. We identified three resistance patterns and six plasmid profiles. Each plasmid profile was characterized by the presence of two large plasmids of 150-180 Kbp. Approximately 60% of strains harbored three or four small plasmids of 1.3 to 9.5 Kbp. The plasmids of 8.5 Kbp encoded resistance to beta-lactam antibiotics and were non-conjugative. The other small plasmids were cryptic and also non conjugative. Salmonella enteritidis isolates were susceptible to many antibiotics, except Tetracycline and Trimethoprim-Sulfamethoxazole. We identified three different resistance patterns but nine plasmid profiles. All plasmid profiles were characterized by the presence of a large plasmid (> 100 Kbp). The number and the diversity of small plasmids were higher than in S. typhimurium strains. There was no parallelism between resistance and plasmid profile: for the same resistance pattern a number of two or three plasmid profiles were found. Our conclusions are that Salmonella typhimurium strains were multiresistant to antibiotics and that many genetically different strains of Salmonella typhimurium and Salmonella enteritidis were responsible for gastroenteritis in children from Cluj County. The increasing antibiotic resistance highlights the need for more refined methods in genetic and epidemiological characterization of bacteria involved in gastrointestinal infections. PMID- 9745326 TI - Phage typing, an accessible and useful method in the epidemiological surveillance of diphtheria. AB - Within the framework of the measures for diphtheria prevention and control, phage typing of isolates is useful for epidemiological investigations in a disease focus and in general population. The identification of asymptomatic carriers of toxigenic C.diphtheriae and its eradication decrease the transmission rate of the disease. The study of the circulating phage types (6058) enabled us to demonstrate the frequency and distribution of the different phage types, toxigenic and nontoxigenic in Romania. With the current knowledge of the circulating phage types it can be concluded that imported cases of diphtheria have not occurred so far. The Romanian phage typing schemes are able to type isolates from overseas, sometimes through phage adaptation methods. Our phage typing schemes are also able to demonstrate any modifications in the phage sensitivity patterns of diphtheria. PMID- 9745327 TI - Streptococcal erythrogenic toxin spe A gene detection by polymerase chain reaction in strains isolated in Albania. AB - The presence of gene encoding erythrogenic toxin type A (spe A) was determined by the polymerase chain reaction (PCR) to target specific sequences in 72 strains of Streptococcus pyogenes representative T type strains, which were associated with scarlet fever, impetigo, tonsillitis, isolated between the years 1980-1982 and in 1995 by carriers. Isolates showed statistically significant differences in the presence of spe A. With scarlet fever the strains had a 22.22% association, with impetigo had a 8.33% association. With tonsillitis' and with carriers had a low association, 5.55% and 6.9% respectively. The analysis of the data indicated that strains with certain T type surface antigens showed a higher (such as T-1, T-2, T 5) or lower (such as T-4, T-13, T-27) tendency to contain the spe A gene were more likely to be associated with scarlet fever and impetigo than with other types of diseases. PMID- 9745328 TI - The immune response induced by a split influenza vaccine. Studies on cellular immunity. AB - The aim of this study is to evaluate the effects exerted in vivo upon the general reactivity of the immune system by a commercial split influenza vaccine produced by the Cantacuzino Institute, Bucharest-Romania. The vaccine was intramuscularly administered to 14 volunteers with no precedent of influenza vaccination and no major immune disorders. We have investigated in vitro the polyclonal proliferation of peripheral lymphocytes and the phagocytosis developed by peripheral granulocytes, before and three weeks after vaccination. Our experimental results indicate that the vaccine might have a modulatory action on peripheral leukocyte concentration induces the activation of polyclonal lymphocyte proliferation and of the phagocytosis potential of granulocytes. These effects are not dependent on the age of the vaccinees. PMID- 9745329 TI - Cytomegalovirus (CMV) in cervical secretion and breast milk. A thirty years perspective. AB - During the past 30 years prospective epidemiologic studies have clearly established that infants commonly acquire CMV infection in the immediate perinatal or early postnatal period. CMV reaches the offspring with uterine cervical secretions during the birth process and/or with maternal milk during the breast-feeding period, usually resulting in asymptomatic infection in full-term infants. In young women cervical shedding of CMV may reflect a pelvic inflammatory disease and involves the risk of sexual transmission. PMID- 9745330 TI - Isolation and speciation of Prevotella strains from periodontal abscesses. AB - The aims of the study were to isolate and to identify at species level the Prevotella strains in pus samples collected by needle aspiration from 25 Romanian patients with periodontal abscesses. Gram-stained smears and cultures on selective and nonselective media were performed from each of the 25 pus samples. The isolates were identified on the basis of Gram staining, cultural characteristics and standard biochemical reactions. The Gram-negative anaerobic bacilli isolates were biochemically characterized and identified at species level using the Rapid ID 32 A system (Bio Merieux, France). Fifteen Prevotella isolates belonging to one of the following species: P. melaninogenica, P. denticola, P. oralis, P. loescheii and P. bivia were recovered. All Prevotella isolates reacted similarly in 20 tests in the Rapid ID 32 A system. The P. melaninogenica strain showed approximately the same biochemical profile and only two sugar fermentation tests were not constantly positive. The study confirmed that Prevotella is often involved in periodontal abscesses (> 50% of the cases) in association with other anaerobic or/and aerobic bacteria. P. melaninogenica was the most frequently isolated Prevotella species from the investigated cases. PMID- 9745331 TI - Study of the leptospirotic etiology in the digestive pathology, particularly in acute abdomen. AB - The paper approaches the possible leptospirotic etiology of non-icteric or late icteric digestive manifestations: digestive hemorrhage, particularly epigastric, acute cholecystitis (non-lithiasic), acute appendicitis, acute gastroenterocolitis, etc., which can be indicative of an acute or latent (from 14 days to several months) leptospirosis. A number of 300 patients with high fever, epidemiological data relevant for leptospirosis and diagnosis upon admission to hospital were investigated using the work method presented in the paper. The synoptical table presents in detail 30 serologically confirmed leptospirosis cases; this reveals the fact that any serotype can be determining and the severe evolution may occur when the etiological diagnosis and treatment are delayed. The 1.67% incidence for the cases clinically expressed by the studied pathology points to the need to consider this pathology as a possible expression form of leptospirosis against the specific epidemiological context. PMID- 9745332 TI - Procain and diethylaminoethanol influence on the release of free oxygen radicals by polymorphonuclear leukocytes, in rabbits and humans. AB - The investigations were conducted on 3 groups of New Zealand rabbits: 1) controls; 2) injected with procain, i.m. 15 mg/kg body weight, daily, for 30 days; 3) injected with diethylaminoethanol (DEAE), 15 mg/kg body weight, daily, for 35 days. The study was made also on human leukocytes, isolated from the peripheral blood of 10 clinically healthy subjects (adults), procain and DEAE action being investigated in vitro. The free oxygen radicals (FOR) released by PMN leukocytes were evaluated by chemiluminescence, in vitro. Addition of procain or DEAE had no effect on the release of FOR by PMN leukocytes of control rabbits. In the experiment made on rabbits treated with procain or DEAE, the release of FOR by PMN leukocytes was much more reduced, as compared to controls. In the rabbits treated with procain, the intensity of the emitted light was 2.27 mV, in those treated with DEAE, 3.46 mV, while in the controls, the mean value was 6.74 mV. In the in vitro experiments performed on human PMN cells stimulated with opsonized zymosan (OZ), addition of procain or DEAE had an inhibiting effect on the FOR release. As compared to control, the means of the FOR values decreased from 59 to 41.2 mV in case of procain addition and from 67.7 to 50 mV in case of DEAE addition. The fact that the inflammation is associated with accumulation of free radicals, suggests the opportunity to test these substances, especially DEAE, as antioxidant agents. PMID- 9745333 TI - Expression of activation surface markers, interleukin-2 synthesis and apoptosis rate in fresh or cultured lymphocytes from HIV-infected children. AB - In 2 groups of HIV-infected children aged from 7 to 10 years (ARC and AIDS, respectively) the following immune markers: % expression of CD69, CD25, HLA-DR activation surface determinants, IL-2 synthesis, rate of apoptosis were tested in non stimulated, PHA-stimulated or PHA and IL-12-stimulated T cell cultures. In all HIV-originated cells a decrease of CD69 expression and an increase of CD25 and HLA-DR expression were found. A strong correlation could be noticed between the clinical stage of the AIDS infection and the in vitro IL-2 production and the percentages of apoptotic cultured cells. IL-12 supplementation of PHA-stimulated cell samples restored the IL-2 synthesis and reduced the apoptosis rates only in the ARC-group, but not in the AIDS-group. The significance of the present data in the clinical and therapeutic monitoring of HIV infection among low-aged people are discussed. PMID- 9745334 TI - Antidiphtheria and antitetanus immunity in various age groups (0-70 yrs) in Romania. AB - The evaluation of the antidiphtheria and antitetanus immunity level is still a topical question. The paper approaches the study of antidiphtheria and antitetanus immunity on age groups in 3,768 and, respectively, 4,001 subjects. In the studied population, the results of estimating immunity revealed an antidiphtheria percentage of 87.16% and a protection level against tetanus of 94.47%. The immunity level against both diphtheria and tetanus varied depending upon the studied age. The highest susceptibility values against diphtheria (29.42%) were recorded in the 6th decade and against tetanus (18.89-28.27%) in the 6th and, respectively, 7th decade. A high antidiphtheria and antitetanus immunity level (over 90%) was reported in children and young people after the IIIrd and IVth boosters). PMID- 9745336 TI - Mycotic vaginitis caused by slow growth fungi. AB - The bacteriological and mycological investigations performed in a female diabetic patient, aged 72, with genital secretion and local discomfort revealed slowly growing fungi, identified as Saccharamyces heterogenicus in the cervix uteri secretion. The cultures performed on Sabouraud medium were positive after 48 hrs incubation in liquid medium and after 4 days in solid medium. The strain was sensitive to: Clotrimazole, Diflucan, Nizoral, Nystatin. PMID- 9745335 TI - The response in hemagglutinoinhibiting antibodies following influenza vaccination of HIV-infected children. AB - The study investigated the response in hemagglutinoinhibiting antibodies (HI) induced by the purified inactivated trivalent influenza vaccine prepared for the epidemic season 1996-1997, administered to a group of 24 human immunodeficiency virus (HIV)-infected children as compared to a group of HIV seronegative controls. The titres of serum HI antibodies were determined before and 30 days after vaccination. The evolution of the immune response in HIV seropositive children showed significant increases against all the antigenic components of influenza vaccine both as concerning the geometrical mean titres of HI antibodies and the protection rate (titres > or = 1:40). However, the amplitude of the immune response reported in the HIV seropositive group was lower, but insignificantly as compared to the HIV seronegative control group. No association between the individual response in HI antibodies and the determined amount of CD4' T lymphocytes was noticed. PMID- 9745337 TI - The formation of frequent additional colonies and of sectors (areas) at fungi. AB - By using single-spore culture method in three points standard system, additional colonies and sectors (areas) are formed due to the dislocation of growing colonies and dissemination of conidia, respectively. Droplets (resulting from condensation of the exudate on Petri dish lids) falling on or touching the conidial masses are responsible for sectors (areas) and additional colonies formation. PMID- 9745338 TI - The mosquitoes (Diptera: Culicidae) in the area of the middle course of the river Somesul Mare (Romania): faunistical and ecological data. AB - The larval and adult mosquito populations in the locality Salva and its surroundings on the middle course of the river Somesul Mare were investigated from March to October 1994. 17 mosquito species (11 as larvae and 13 as adults) were recorded: Aedes cataphylla, Aedes cantans Aedes leucomelas, Aedes dorsalis, Aedes caspius, Aedes vexans, Aedes flavescens, Aedes cinereus, Aedes annulipes, Aedes geniculatus, Culex modestus, Culex pipiens, Culex territans, Culiseta alaskaensis, Anopheles maculipennis s.l., Anopheles claviger, and Anopheles plumbeus. An ecological analysis of the mosquito populations regarding the diversity, dominance, the frequency and succession of the species during their monthly dynamics was performed. PMID- 9745339 TI - [Homeopathy--possibilities and limits]. PMID- 9745340 TI - [Anthroposophic expansion of medicine]. PMID- 9745341 TI - [Ayurvedic medicine--possibilities and limits]. PMID- 9745342 TI - [Significance of music in modern medicine]. PMID- 9745343 TI - [Creative (psycho-) therapy--art therapy]. PMID- 9745344 TI - [Analysis of the unfolded protein response: an intracellular signaling pathway from the endoplasmic reticulum to the nucleus]. PMID- 9745345 TI - [Regulation of bone resorption by hormone and cytokine]. PMID- 9745346 TI - [Recent advances in the analysis of ganglioside synthesis]. PMID- 9745347 TI - [Structural and functional analysis of gp64, a membrane protein of the cellular slime mold Polysphondylium pallidum]. PMID- 9745348 TI - [New targets of rho GTPases, mDia and Bnilp, and formin homology protein family]. PMID- 9745349 TI - [Nociceptin and nociceptin receptor]. PMID- 9745350 TI - [Secretion defect of kininogens and bulk flow]. PMID- 9745351 TI - [Novel chemokines identified through bioinformatics]. PMID- 9745352 TI - [On the "folkloric" psychotherapy to schizophrenic patients--based on phenomenological study]. AB - Psychotherapeutic treatment of schizophrenia is generally considered difficult. One reason for this is that the doctor and patient can easily fall into a relationship of conflict with each other concerning the propriety of "judgments which are morbidly and mistakenly made (K. Jaspers)", referred to as delusions. We carried out close phenomenological structure-analyses of the delusions and of patients' fundamental experiences, based on the premise that a patient with delusions probably has some actual grounding for these in the patient's own concepts, considering the fact that the patient firmly believes these delusions. As a result, we have clarified the following matters from the primary experience of delusions. 1) We found that patients are in a conflicted mental condition which can be considered a collapse of adaptability to "Seken". 2) In this condition of conflict, patients feel guilt relative to "Seken" or feel that they are indebted and should be punished. When patients complained of their primary experience, we were able to persuade them to reserve their judgment of their primary experience, by 3) having each patient listen to the folktale "Torikuyou" in which the "logic of stealing" and the "logic of being stolen", appear in a reciprocal relationship relative to the constitution of crime and punishment, by 4) explaining to each patient about the ambiguity and reciprocity of reality experienced, 5) instead of disputing the propriety of patient's judgment about primary experience, doctor and patient worked together to enable the patient to form a positive understanding of the primary experience. 6) We reduced the patient's psychological conflict relative to primary experience, and were able to defuse and distance the patient's delusions caused by erroneous judgment of primary experience. 7) Regarding the area in which this type of psychotherapeutic approach shows efficacy, we analyzed the concept of "Seken" as a world which can cause conflicts relative to primary experiences. 8) We also analyzed "Giri" as a norm of "Seken" from which patients misconceive that they have deviated, in addition, 9) from the viewpoints of anthropology and cultural anthropology, we analyzed the bases for "Kotowaza (proverbs)" and "Monogatari (folktale)" such as "Torikuyou", which themselves can show psychotherapeutic efficacy. We consider that the psychotherapeutic approach has previously been developed around the concepts of the "individual" and "society", but we made our psychotherapeutic approach from the concept of "Seken" (yononaka = hito: person) that is a structure with deep strata of tradition and culture in Japan, and have reported its concrete development through the presentation of 3 typical cases of schizophrenia with difficulty in adapting to society due to showing the delusion of persecution in their foreground. PMID- 9745354 TI - [Psychopathological study on schizophrenic autism through the paintings of a case of simple schizophrenia]. AB - Autism is the symptom which has the most specific features of schizophrenia. However, the content of pathological experience of the patients has not yet been clarified as it was never told by themselves. In the present study, a case of simple schizophrenia with a major symptom of autism is reported. Schizophrenic autism was studied psychopathologically by drawing tests. The analysis of drawings was summarized by the following characteristics. 1. The objects were drawn so as to be small at the center. The composition was further characterized by the overwhelmingly predominant empty space of its circumference, which we termed "reversed zoom lens effect". This seemed to show that the objects were isolated and removed from the patient. 2. The entire image of the theme was not drawn. 3. Lack of vitality was observed in all the drawings. On the basis of the above characteristics of these drawings and the clinical findings, the following were suggested as the pathology of the patient's experience: 1. The "reversed zoom lens composition" seen in the drawings suggested that the psychological distance between objects and the patient might be expanded. 2. The patient was alienated from the objective world to which he had once been accustomed as the object lacked vitality and familiarity. 3. The pathology of schizophrenic autism observed in the patient could be expressed as "alienation from objective experience." 4. It was considered that although akin to depersonalization, "alienation from objective experience" was the pathology characteristic of schizophrenia. PMID- 9745353 TI - [A study of neuroleptic malignant syndrome in the presenium and senium]. AB - Recently, with the increase in elderly population, we have had more opportunities to administer neuroleptics to elderly patients for hallucinatory delusional state, delirium, psychomotor excitement, wandering etc. However, little is known about the characteristics of the neuroleptic malignant syndrome (NMS) in elderly patients, which is the most serious side effect of neuroleptics. In this paper, we present the clinical course of five NMS patients in the presenium and senium. Case 1 was 72-year-old male who was diagnosed as having dementia of Alzheimer's type (with late onset). He showed nocturnal wandering, insomnia, and irritability. Tiapride 60 mg per day had been administered previously. Just after the addition of oxypertine 10 mg per day, NMS occurred, and he died of pneumonia a week later. Case 2 was 75-year-old male who was diagnosed as having vascular dementia. He showed insomnia, hyperactivity and wandering. He had been given levomepromazine (LPZ) 10 mg per day over a long period of time. At first, he had daily episodic fever, however, serum CPK levels did not increase at that time. A month later, all the symptoms of NMS appeared and then the patient's condition suddenly deteriorated and he died three days later. Case 3 was a 64-year-old male who was diagnosed as having dementia of Alzheimer's type (with early onset). He showed insomnia, irritability and violence. Tiapride 50-125 mg per day was administered along with oxypertine 50-115 mg per day. Almost two months later, NMS occurred. He had daily episodic fever at first, extrapyramidal symptoms and autonomic instabilities gradually increased. Soon after symptoms of NMS were completed. In this case, NMS seemed to be induced by bacterial pneumonia after long term administration of LPZ 5 mg per day. Case 4 was a 75-year-old female who was diagnosed as having dementia of Alzheimer's type (with late onset). She showed hallucinatory delusional state. Although she had autonomic instabilities just after adminstration of haloperidol 1-2 mg per day, NMS itself occurred after discontinuing the neuroleptic. Case 5 was a 61-year-old female who was diagnosed as having schizophrenia at the age of forty. She was given various neuroleptics over a period of time. The neuroimaging in SPECT showed her cerebral cortex was generally hypoactive. She had a tendency to have autonomic instabilities after the administration of relatively high potential neuroleptics. Risperidone 3-6 mg per day was administered, and almost a month later, autonomic instabilities increased and she was diagnosed as having NMS. All the patients would be able to have brain dysfunction, which suggested that such patients may be liable to NMS. In our patients, NMS occurred after the additional administration of oxypertine 10 mg per day or after long time administration of LPZ 5 mg per day. It was suggested that NMS could occur after the administration of low dose and relatively low potential neuroleptics in elderly patients. Our 3 of 5 patients showed the delayed type of NMS, which might be relatively more frequent in senior and presenior patients than in younger patients. In case 3, NMS was induced by the somatic disease (bacterial pneumonia), however in other cases, NMS was not always induced by somatic disease. Our 4 of 5 patients experienced some of the symptoms of NMS--episodic fever, extrapyramidal symptoms and autonomic instabilities--before the onset of NMS. Such symptoms may be "pre-steps" to NMS. Once NMS occurred, the patient's systemic condition tended to deteriorate acutely. Due to the fact that our 2 of 5 patients died, it was suggested that the prognosis of the NMS patients in presenium and senium tends to be much worse. It is important to find the "pre-steps" to NMS and treat them as soon as possible for better prognosis. PMID- 9745355 TI - Calcitonin gene-related peptide--a new concept in receptor-ligand specificity? PMID- 9745356 TI - Receptor targets in William Tell country: insights into receptor biology using fluorescent proteins. PMID- 9745357 TI - Involvement of T cells in drug-induced allergies. PMID- 9745358 TI - 5-HT1-like receptors: a time to bid goodbye. AB - It is exactly half a century ago that 5-hydroxytryptamine was discovered and over four decades ago two types of 5-HT receptors were described. In this article, Pramod Saxena, Peter De Vries and Carlos Villalon trace the development of the modern classification and nomenclature of 5-HT receptors, which now include more than a dozen subtypes. In doing so, they advocate that the so-called '5-HT1-like' receptors, having been shown to be a heterogeneous population of 5-HT1B, 5-HT1D and 5-HT7 receptors, are now redundant. PMID- 9745359 TI - Induction of cytokine receptors by glucocorticoids: functional and pathological significance. AB - Current concepts on the role of glucocorticoid hormones in the regulation of inflammatory and immune responses depict this role as being inhibitory. Over the past decade, however, a large variety of studies have shown that glucocorticoids also exert stimulatory effects on immune function, suggesting that the present concept of the role of glucocorticoids in the immune system in not sufficient and needs to be extended. Here, Jan Wiegers and Hans Reul ask how these apparently paradoxical effects fit together and what their functional and pathological significance might be. PMID- 9745360 TI - Regulation of the expression and function of the M2 muscarinic receptor. AB - Since the cloning and expression of many of the G protein-coupled receptors during the 1980s, there has been a massive increase in our understanding of many aspects of their function. The use of molecular biology to engineer and express mutant receptors has made it possible to determine key amino acids involved in receptor function. Although advances in molecular biology have contributed greatly to our understanding of the pharmacology and structure of the five subtypes of muscarinic receptor, much remains to be learned about the factors that regulate their expression and function. This review by El-Bdaoui Haddad and Jonathan Rousell describes the current state of awareness and highlights recent advances made in the elucidation of the mechanisms involved in muscarinic receptor regulation. Because most is known about the regulation of expression of the M2 receptor subtype, particular attention will be paid to it. Furthermore, this receptor subtype plays an important role in regulating acetylcholine output from airway cholinergic nerves, and there is substantial evidence from studies both in vivo and in vitro in human and animal models that these receptors are dysfunctional in asthma. PMID- 9745361 TI - Glutamate transporters are oxidant-vulnerable: a molecular link between oxidative and excitotoxic neurodegeneration? AB - Increasing evidence indicates that glutamate transporters are vulnerable to the action of biological oxidants, resulting in reduced uptake function. This effect could contribute to the build-up of neurotoxic extracellular glutamate levels, with major pathological consequences. Specific 'redox-sensing' elements, consisting of cysteine residues, have been identified in the structures of at least three transporter subtypes (GLT1, GLAST and EAAC1) and shown to regulate transport rate via thiol-disulphide redox interconversion. In this article, Davide Trotti, Niels Danbolt and Andrea Volterra discuss these findings in relation to the emerging view that in brain diseases oxidative and excitotoxic mechanisms might often operate in tight conjunction to induce neuronal damage. In particular, they review evidence suggesting a possible involvement of oxidative alterations of glutamate transporters in specific pathologies, including amyotrophic lateral sclerosis, Alzheimer's disease, brain trauma and ischaemia. PMID- 9745362 TI - Airway epithelium: more than just a barrier! AB - Airway hyper-responsiveness and epithelial cell damage are associated commonly with asthma. The airway epithelium is a physical barrier that protects sensory nerves and smooth muscle from stimulation by inhaled irritants. In addition, epithelial cells release mediators that can inhibit bronchoconstriction by relaxing the underlying smooth muscle: so-called 'epithelium-derived relaxing factors' (EpiDRFs). Clear functional evidence for EpiDRFs is provided by experiments where different endogenous mediators induce the relaxation of tracheas containing epithelium, but cause a contraction in preparations lacking this layer. Here, Gert Folkerts and Frans Nijkamp describe the pharmacological relevance of the putative EpiDRFs, prostaglandin E2 and NO, in the modulation of airway tone under basal conditions in vitro and in vivo. Special attention is paid to the role of both EpiDRFs in the development of airway hyper responsiveness in animal models and in patients with asthma. PMID- 9745363 TI - [Public health policy significance of life style medicine]. PMID- 9745364 TI - [New information on so-called whiplash cervical trauma]. AB - Whiplash injuries of the cervical spine probably are much too often complained of, certified and compensated. Many of these injuries arise from suggestion and/or deliberate simulation. The difficulty of verifying minor whiplash injuries is well known and used widespread, to try gaining compensation from the insurance companies. In accidents with a collision speed of less than 20 km/h (obvious damage to the car) it is unlikely to suffer an structural injury of the cervical spine. Even accidents with a higher collision speed do not necessarily lead to a whiplash injury. An interval without symptoms is a hint against impairment. If it is not possible to get reproducible evidence of an injury caused by the accident this must clearly be stated. Therapy should impose early return to normal life style, a soft collar is obsolete and sick leave is in contradiction to the routine rarely needed. The current way how the insurance companies compensate encourages the symptoms and needs to be revised. PMID- 9745365 TI - [Risk assessment expanded accident insurance for children]. AB - Disability is a well known and tragic event for children. While adults are an established group for specific disability insurance cover, children were often neglected in the past. Although parents, organizations and paediatricans are aware of the risk, children specific incidence rates for disability are hardly available. The only sufficient source for some statistical data are the accident statistics because they represent a substantial group of specific cause related disability for children. Incidence rates for disease related chronic severe impairment or disability in children are either derived by single disease research or actuarial calculation of the German Social Disability Registration. Based on this statistical background, an extended accident insurance for children was introduced in Germany covering both accidents and disabling diseases. The key limitation for all variations of this insurance are exclusion clauses for congential diseases and mental disorders. This insurance requires a new approach in underwriting of the health risks. Because of the substantial number of impaired children, a simple decline of substandard cases are unacceptable. The early experience or medical underwriting shows predominantly health impairments of the following types: allergies, bronchial asthma, ectopic eczema (neurodermitis), disorders of speech and articulation, vision disorders and mental impairments. The suggested solution for underwriting of substandard risks is the predetermination of the possible future maximum degree of disability. The need for underwriting guidelines is supported by the market impact of the new disability cover with thousands of insurance policies issued in the first month after introduction. PMID- 9745366 TI - [Oxygen treatment methods--a critical analysis of established and controversial procedures (I)]. PMID- 9745367 TI - [Chronic backache--do guidelines/standards exist?]. AB - Chronic (low) back pain patients are about to become a main problem of epidemiological and social aspects, especially concerning working conditions. Statistical investigations show rapid progression in application for disability pensions. The author wants to point out that statistical normal phenomena as (low) back pain can no longer be explained by our so-called established handling in reasons, diagnosis and treatment, even not in prophylaxis. Therefore there is a great need for--mainly bio-socio-psychological--studies and efforts concerning etiology, diagnosis and therapeutic concepts of low back pain, especially focussed on working place conditions and disability. In the uplined paper some new concepts are presented and discussed in order to trigger some new points of view and make clear, that further investigations are necessary. PMID- 9745368 TI - [Are so-called alternative therapeutic procedures in pain treatment the solution?]. AB - So-called alternative methods are surely not the solution for the therapy of chronic pain. Although those methods may often produce a marked placebo effect, it must be pointed out that so far there has not been any proof of a specific effect. At some methods, especially acupuncture, a therapeutic effect seems possible. Further carefully planned clinical studies are necessary. Finally the therapy with such unproofed methods often causes high costs that contrast to the effectivity. PMID- 9745369 TI - [(Incipient) paradigm change in cardiology?]. PMID- 9745370 TI - [Comment on W. Hausotter: Fibromyalgia--a dispensable disease concept?]. PMID- 9745371 TI - Expanded Programme on Immunization (EPI). Standardization of the nomenclature for describing the genetic characteristics of wild-type measles viruses. PMID- 9745372 TI - The spectre of inbreeding in the early investigation of heredity. AB - Inbreeding introduced by R. Bakewell (1725-1795) in England for creating new animal races, was opposed by animal breeders on the Continent on religious grounds, and was soon introduced in sheep breeding for wool production in Moravia. In 1790-1840 the protagonists repeatedly rejected 'the spectre of inbreeding' and included consanguineous matching in scientific breeding. In 1836 they even formulated the research question of heredity and next year proposed the inductive method for its investigation. The achievements of sheep breeders instigated German breeders to reject the dogma of the constancy of race and to elaborate the theory of individual potency. Treating heredity as the force under the influence of environment they could not solve the enigma. The question formulated in 1836 was explained in 1865 by Gregor Mendel. His theory was not perceived by animal breeders as well as by biologists up to the end of the century. PMID- 9745373 TI - [Pathology and politics--Ludwig Aschoff (1866-1942) and the German way in the Third Reich]. AB - This paper focuses on a prominent figure in medicine, one who had no visible political ambitions in terms of the NS-regime: Ludwig Aschoff (1866-1942), pathologist at the University of Freiburg from 1906 to 1936. The period between 1914 and 1933 is emphasized in order to show the development of his political and scientific ideas, which were intertwined from the beginning: Aschoff admired the Kaiserreich, and in World War I he tried to carry out a socio-political programme in that he examined the constitution of the German Volk by performing autopsies of almost every German soldier who had been killed in action. After 1918 he retained his conservative ideas and his medical programme which meant concentrating mainly on constitutional ideas. Although Aschoff tried to reestablish international cooperation and to support the new Republic, he ultimately did support anti-democratic ideas. This attitude proved to be a handicap with regard to his reaction to National Socialism in 1933: Although he did not become a member of the party and a strong supporter, he welcomed the new regime. Aschoff, without being aware of his role, supported the acceptance of the regime within the medical scientific community. It will be possible to obtain deeper insight into the NS-period by encouraging research on its 'mediators', for example, Ludwig Aschoff. PMID- 9745374 TI - The transformation of molecular biology on contact with higher organisms, 1960 1980: from a molecular description to a molecular explanation. AB - The convergence of developmental biology--embryology--and molecular biology was one of the major scientific events of the last decades of the twentieth century. The transformation of developmental biology by the concepts and methods of molecular biology has already been described. Less has been told on the reciprocal transformation of molecular biology on contact with higher organisms. The transformation of molecular biology occurred at the end of a deep crisis which affected this discipline in the sixties and seventies and which led to a cruel criticism of the preexisting models of gene regulation. Numerous new, sometimes heterodox, models were proposed to describe the level at which gene regulation took place and its underlying mechanisms. The crisis resolved itself at the beginning of the eighties with the rapid accumulation of results from genetic engineering techniques and, above all, a displacement of the descriptive level from the molecule to the cell. This displacement gave molecular biologists the 'explanandum' which had been cruelly lacking during their initial study of higher organisms. The new molecular cell biology is an interfield explanation of living phenomena, relating a description and an interpretation localized at different levels of organization. PMID- 9745376 TI - Discourses of mood and depression. PMID- 9745375 TI - A cold war colonial science: the Atomic Bombing Casualty Commission's study of genetic mutations in the children of atomic bomb survivors. PMID- 9745377 TI - Local and relational aspects of face distinctiveness. AB - Distinctiveness contributes strongly to the recognition and rejection of faces in memory tasks. In four experiments we examine the role played by local and relational information in the distinctiveness of upright and inverted faces. In all experiments subjects saw one of three versions of a face: original faces, which had been rated as average in distinctiveness in a previous study (Hancock, Burton, & Bruce, 1996), a more distinctive version in which local features had been changed (D-local), and a more distinctive version in which relational features had been changed (D-rel). An increase in distinctiveness was found for D local and D-rel faces in Experiment 1 (complete faces) and 3 and 4 (face internals only) when the faces had to be rated in upright presentation, but the distinctiveness of the D-rel faces was reduced much more than that of the D-local versions when the ratings were given to the faces presented upside-down (Experiments 1 and 3). Recognition performance showed a similar pattern: presented upright, both D-local and D-rel revealed higher performance compared to the originals, but in upside-down presentation the D-local versions showed a much stronger distinctiveness advantage. When only internal features of faces were used (Experiments 3 and 4), the D-rel faces lost their advantage over the Original versions in inverted presentation. The results suggest that at least two dimensions of facial information contribute to a face's apparent distinctiveness, but that these sources of information are differentially affected by turning the face upside-down. These findings are in accordance with a face processing model in which face inversion effects occur because a specific type of information processing is disrupted, rather than because of a general disruption of performance. PMID- 9745378 TI - The effect of age of acquisition on speed and accuracy of naming famous faces. AB - Three experiments examined whether famous faces would be affected by the age at which knowledge of the face was first acquired (AoA). Using a multiple regression design, Experiment 1 showed that rated familiarity and AoA were significant predictors of the time required to name pictures of celebrities' faces and the accuracy of producing their names. Experiment 2 replicated an effect of AoA using a factorial design in which other attributes of the celebrities were matched. In both Experiments 1 and 2, several ratings had been collected from participants before naming latency data were collected. Experiment 3 investigated the accuracy and latency of naming celebrities without any prior exposure to the stimuli. An advantage for naming early acquired celebrities was observed even on the first presentation. The participants named the same celebrities in three subsequent presentations of the stimuli. The effect of AoA was not significant on the fourth presentation. The implications of these results for models of face naming and directions for future research are discussed. PMID- 9745379 TI - The role of implicit memory in controlling a dynamic system. AB - The relationship between implicit memory and implicit learning is explored. Dienes and Fahey (1995) showed that learning to control a dynamic system was mediated by a look-up table consisting of previously successful responses to specific situations. The experiment reported in this paper showed that facilitated performance on old situations was independent of the subjects' ability to recognize those situations as old, suggesting that memory was implicit. Further analyses of the Dienes and Fahey data replicated this independence of control performance on recognition. However, unlike the implicit memory revealed on fragment completion tasks, successful performance on the dynamic control tasks was remarkably resilient to modality shifts. The results are discussed in terms of models of implicit learning and the nature of implicit memory. PMID- 9745380 TI - Irrelevant sound disrupts order information in free recall as in serial recall. AB - The claim that the sensitivity of free recall to disruption by irrelevant sound is a function of the extent to which rote rehearsal is employed as a mnemonic strategy was investigated in two experiments. The degree of disruption by irrelevant sound in terms of both item and order information was contrasted under serial and free recall instructions. Irrelevant sound was found to disrupt order and item information equally in serial and free recall tasks (Experiment 1). Contrary to previous reports, an effect of irrelevant sound was also demonstrated on free recall of particularly long lists, and the interaction between list length and retention interval in the irrelevant sound effect was examined (Experiment 2). Generally, the results support the view that irrelevant sound disrupts the use of order cues. PMID- 9745381 TI - Automatic spatial updating during locomotion without vision. AB - People can update their spatial relationships relative to the environment while walking without vision. The hypothesis that such updating is automatic was tested in a locomotor task in which the subjects were asked to refrain from updating their positions. Subjects walked without vision to one of four previously seen targets via a second location. In one condition--the updating condition--the subjects were asked to walk to the real position of the target relative to the second location; in another--the ignoring condition--they were asked to imagine that they had not moved from the starting point and to walk from the second location as if walking to the target from the initial location. When the subjects were asked to start walking to the target as soon as it was named by the experimenter, they performed better in the updating condition than in the ignoring condition. When the subjects were allowed more time to respond, the difference in performance between these two conditions disappeared. The results suggest that the subjects automatically updated their positions as they moved, but that, given enough time, they could override this updating retrospectively using more deliberate cognitive processing. PMID- 9745383 TI - Perceptual learning and free classification. AB - Two experiments are reported that investigate the effects of stimulus preexposure on discrimination performance in a free classification task, using adult humans as subjects. In free classification subjects are asked to put stimuli into groups in any way that seems reasonable or sensible to them. Experiment 1 shows that the effect of preexposure is contingent on stimulus structure. Experiment 1b is the first demonstration of a retardation in learning as a consequence of simple preexposure in adult human subjects (previous demonstrations have relied on incidental or masked preexposure). Experiment 2 further supports the conclusions of Experiment 1 and extends them with the demonstration that stimulus similarity is a crucial factor. Taken together, these experiments rule out a class of attention-based explanations of the phenomena reported here. The experiments also provide novel information about the effects of preexposure. Preexposure can change the actual classifications subjects form in addition to altering the rate at which they are formed. Implications of these results for current theories of category formation and perceptual learning are considered. PMID- 9745382 TI - Peak procedure performance in young adult and aged rats: acquisition and adaptation to a changing temporal criterion. AB - Twenty-four-month-old and 4-month-old rats were trained on a peak-interval procedure, where the time of reinforcement was varied twice between 20 and 40 sec. Peak times from the old rats were consistently longer than the reinforcement time, whereas those from younger animals tracked the 20- and 40-sec durations more closely. Different measures of performance suggested that the old rats were either (1) systematically misremembering the time of reinforcement or (2) using an internal clock with a substantially greater latency to start and stop timing than the younger animals. Old rats also adjusted more slowly to the first transition from 20 to 40 sec than did the younger ones, but not to later transitions. Correlations between measures derived from within-trial patterns of responding conformed in general to detailed predictions derived from scalar expectancy theory. However, some correlation values more closely resembled those derived from a study of peak-interval performance in humans and a theoretical model developed by Cheng and Westwood (1993), than those obtained in previous work with animals, for reasons that are at present unclear. PMID- 9745384 TI - Chiropractic care for back pain. PMID- 9745385 TI - A thyroid dilemma. PMID- 9745386 TI - Coping with Crohn's disease. PMID- 9745388 TI - A complex pain. PMID- 9745387 TI - Disrobing in the doctor's office. PMID- 9745389 TI - Hormone use of questionable benefit in women with heart disease. PMID- 9745390 TI - Parrot fever. PMID- 9745391 TI - Minimizing breast cancer's spread. PMID- 9745393 TI - Can antireflux therapy improve asthma? PMID- 9745392 TI - Living environment affects Alzheimer's agitation. PMID- 9745394 TI - Preventing inherited ovarian cancer. PMID- 9745395 TI - Clinical review 97: Syndromes of severe insulin resistance. PMID- 9745396 TI - Primary medical therapy for acromegaly. PMID- 9745397 TI - Octreotide as primary therapy for acromegaly. AB - The effects of octreotide (up to 5 yr) as primary treatment in 26 patients with acromegaly were compared with those in 81 patients with acromegaly who received octreotide as secondary or adjunctive therapy after previous surgery and/or pituitary radiation. These patients were part of a multicenter study that took place between 1989-1995. The study was divided into 3 phases beginning with a 1 month placebo-controlled treatment period followed by a 1-month washout period. In the second phase, patients were randomized to treatment with either 100 or 250 micrograms octreotide, sc, every 8 h for 6 months. Octreotide was then discontinued for 1 month and reinitiated at the lower dose for a total mean treatment duration of 39 months. The dose was titrated by each investigator to improve each patient's individual response, which included improvement in symptoms and signs of acromegaly as well as reduction of GH and insulin-like growth factor I (IGF-I) into the normal range. In the second phase of the study, in which patients were randomized to either 100 or 250 micrograms octreotide, three times daily, mean integrated GH and IGF-I concentrations after 3 and 6 months were equivalent in the primary and secondary treatment groups. During long term open label treatment, mean GH fell from 32.7 +/- 5.2 to 6.0 +/- 1.7 micrograms/L 2 h after octreotide injection in the primary therapy group and remained suppressed for a mean period of 24 months (range, 3-60 months). The mean final daily dose was 777 micrograms. In the patients receiving secondary treatment, mean GH fell from 30.2 +/- 7.6 to 5.6 +/- 1.1 micrograms/L after 3 months and remained suppressed for the remainder of the study (average dose, 635 micrograms daily). Mean IGF-I concentrations fell from 5.2 +/- 0.5 x 10(3) U/L (primary treatment group) and 4.7 +/- 0.4 x 10(3) U/L (secondary treatment group) to a mean of 2.2 +/- 0.3 x 10(3) U/L in both groups after 3 months of open label treatment and remained suppressed. IGF-I was reduced into the normal range during at least half of the study visits in 68% of the primary treatment group and in 62% of the secondary treatment group. Patients whose GH levels fell to at least 2 SD below the baseline mean GH were considered responders. There was no significant difference in the percentage of responders in the primary and secondary treatment groups (70% vs. 61%), nor was there a statistical difference in the mean GH concentrations between the groups. Symptoms of headache, increased perspiration, fatigue, and joint pain were reported at baseline by 46%, 73%, 69%, and 85%, respectively, of patients in the primary therapy group and improved during 3 yr of octreotide treatment in 50-100%. Similarly, these acromegaly related symptoms were reported by 62%, 58%, 78%, and 60% of patients in the secondary therapy group, and improvement was noted in 62-88%. Pituitary magnetic resonance imaging scans were available in 13 of 26 patients in the primary treatment group before and after 6 months of octreotide treatment. Tumor shrinkage was observed in 6 of 13 patients, with reduction in tumor volume greater than 25% in only 3. Of 6 patients with documented tumor shrinkage, IGF-I was reduced into the normal range in 4 patients. Of the 7 remaining patients in whom tumor shrinkage was less than 10%, IGF-I was reduced into the normal range in 4 patients. Of the 7 remaining patients in whom tumor shrinkage was less than 10%, IGF-I was reduced into the normal range in 5 patients. The degree of tumor shrinkage did not correlate with the percent reduction in IGF-I or GH. In summary, octreotide was equally effective in 26 previously untreated acromegalic patients (primary treatment group) and 81 patients previously treated with either surgery or pituitary radiation (secondary treatment group). These observations call into question the current practice of surgical resection of all newly diagnosed GH-secreting pituitary adenomas regardless of the likelihood of cure. (AB PMID- 9745398 TI - Therapeutic controversy: The role of laparoscopic surgery in adrenal disease. PMID- 9745402 TI - Albumin synthesis and bone collagen formation in human immunodeficiency virus positive subjects: differential effects of growth hormone administration. AB - Loss of lean tissue often accompanies human immunodeficiency virus (HIV) infection. Exogenous human recombinant GH (hrGH) has been shown to be beneficial in reversing this wasting. However, catabolic effects of hrGH on muscle protein metabolism have also been reported. Therefore, the responsiveness of other GH sensitive tissues, including bone formation and albumin synthesis, has been examined. Anabolic activity in bone, from serum levels of carboxy-terminal propeptide of type I collagen, was stimulated by 2 weeks of hrGH in controls (56 +/- 15%, P = 0.002), patients with asymptomatic HIV (24 +/- 10%, not significant), patients with AIDS (47 +/- 7%, P < 0.001), and patients with AIDS and > 10% weight loss (21 +/- 12%, P = 0.02). Albumin synthesis, determined from the incorporation of L-[2H5]phenylalanine, was increased in response to hrGH in controls (23 +/- 7%, P < 0.05), HIV+ subjects (39 +/- 16%, P < 0.05), and patients with AIDS (25 +/- 7%, P < 0.01). Patients with AIDS and weight loss, however, did not increase albumin synthesis (-0.6 +/- 12%) in response to hrGH. The results indicate variable anabolic responses to hrGH. Bone collagen synthesis remained sensitive to hrGH, whereas, the anabolic action of hrGH on the synthesis of albumin diminished with severity of disease. However unlike muscle protein synthesis, albumin synthesis was not depressed below basal levels by hrGH. PMID- 9745403 TI - Decreased nitric oxide levels and bone turnover in amenorrheic athletes with spinal osteopenia. AB - Amenorrheic athletes have been likened to postmenopausal women, with low estrogen levels and osteopenia. It has been suggested that estrogen exerts its antiresorptive actions on bone via a nitric oxide (NO)-dependent mechanism. This study investigated whether the mechanism of bone loss in amenorrheic athletes is similar to that of postmenopausal women with reduced NO levels and high bone turnover. Eleven amenorrheic athletes, 15 eumenorrheic athletes, and 10 sedentary controls were studied. Spine and hip bone mineral density was measured using dual energy x-ray absorptiometry. Bone turnover was assessed by biochemical markers of formation (osteocalcin and bone-specific alkaline phosphatase) and resorption (deoxypyridinoline). NO metabolites were measured from 24-h urine samples using a chemiluminescence assay. Spine, but not hip, bone mineral density was reduced in the amenorrheic group, compared with the eumenorrheic (P = 0.0001) and control (P = 0.04) groups. Osteocalcin, bone-specific alkaline phosphatase, and deoxypyridinoline were similar in all groups. NO metabolites were lower in the amenorrheic group, compared with controls (P = 0.035), despite a higher dietary intake of nitrates. Unlike postmenopausal women, amenorrheic athletes do not have raised bone turnover but do have reduced NO metabolites and spinal osteopenia. The results show, however, that reduced NO production is a common denominator in both conditions and further support the importance of NO in estrogen-mediated protection of skeletal mass and strength. PMID- 9745404 TI - Leptin levels in protracted critical illness: effects of growth hormone secretagogues and thyrotropin-releasing hormone. AB - Prolonged critical illness is characterized by feeding-resistant wasting of protein, whereas reesterification, instead of oxidation of fatty acids, allows fat stores to accrue and associate with a low-activity status of the somatotropic and thyrotropic axis, which seems to be partly of hypothalamic origin. To further unravel this paradoxical metabolic condition, and in search of potential therapeutic strategies, we measured serum concentrations of leptin; studied the relationship with body mass index, insulin, cortisol, thyroid hormones, and somatomedins; and documented the effects of hypothalamic releasing factors, in particular, GH-secretagogues and TRH. Twenty adults, critically ill for several weeks and supported with normocaloric, continuously administered parenteral and/or enteral feeding, were studied for 45 h. They had been randomized to receive one of three combinations of peptide infusions, in random order: TRH (one day) and placebo (other day); TRH + GH-releasing peptide (GHRP)-2 and GHRP-2; TRH + GHRH + GHRP-2 and GHRH + GHRP-2. Peptide infusions were started after a 1 microgram/kg bolus at 0900 h and infused (1 microgram/kg.h) until 0600 h the next morning. Serum concentrations of leptin, insulin, cortisol, T4, T3, insulin-like growth factor (IGF)-I, IGF-binding protein-3 and the acid-labile subunit (ALS) were measured at 0900 h, 2100 h, and 0600 h on each of the 2 study days. Baseline leptin levels (mean +/- SEM: 12.4 +/- 2.1 micrograms/L) were independent of body mass index (25 +/- 1 kg/m2), insulin (18.6 +/- 2.9 microIU/mL), cortisol (504 +/- 43 mmol/L), and thyroid hormones (T4: 63 +/- 5 nmol/L, T3: 0.72 +/- 0.08 nmol/L) but correlated positively with circulating levels of IGF-I [86 +/- 6 micrograms/L, determination coefficient (R2) = 0.25] and ALS (7.2 +/- 0.6 mg/L, R2 = 0.32). Infusion of placebo or TRH had no effect on leptin. In contrast, GH secretagogues elevated leptin levels within 12 h. Infusion of GHRP-2 alone induced a maximal leptin increase of +87% after 24 h, whereas GHRH + GHRP-2 elevated leptin by up to +157% after 24 h. The increase in leptin within 12 h was related (R2 = 0.58) to the substantial rise in insulin. After 45 h, and having reached a plateau, leptin was related to the increased IGF-I (R2 = 0.37). In conclusion, circulating leptin levels during protracted critical illness were linked to the activity state of the GH/IGF-I axis. Stimulating the GH/IGF-I axis with GH-secretagogues increased leptin levels within 12 h. Because leptin may stimulate oxidation of fatty acids, and because GH, IGF-I, and insulin have a protein-sparing effect, GH-secretagogue administration may be expected to result in increased utilization of fat as preferential substrate and to restore protein content in vital tissues and, consequently, has potential as a strategy to reverse the paradoxical metabolic condition of protracted critical illness. PMID- 9745405 TI - 3,3'-Diiodothyronine concentrations in the sera of patients with nonthyroidal illnesses and brain tumors and of healthy subjects during acute stress. AB - In this article we describe the development of a highly sensitive, accurate, and reproducible RIA for the measurement of 3,3'-diiodothyronine (3,3'-T2) in human serum and brain tissue. The detection limits were 1.8 fmol/g and 1.5 pmol/L in human brain tissue and serum, respectively. Serum concentrations of 3,3'-T2 were measured in 4 groups of patients with nonthyroidal illnesses (NTI), i.e. brain injuries (n = 15), sepsis (n = 24), liver disease (n = 22), and brain tumors (n = 23). The mean serum concentration of 3,3'-T2 in 62 healthy controls was 46.6 +/- 20.0 pmol/L. 3,3'-T2 levels declined significantly with increasing age. They were significantly lower in patients with brain injury (34.2 +/- 19.4 pmol/L; P = 0.006), were at the upper limit of normal in patients with sepsis (57.0 +/- 36.9 pmol/L; P = 0.06), and were elevated in patients with liver disease (72.6 +/- 56.7 pmol/L; P = 0.04) and brain tumors (89.0 +/- 40.9 pmol/L; P = 0.01). The serum levels of T3 were significantly lower than those in controls in all 4 patient groups. Serum concentrations of 3,3'-T2 were significantly enhanced in 9 patients with hyperthyroidism (85.4 +/- 43.0 pmol/L; P = 0.01) and were reduced in 12 patients with hypothyroidism (14.9 +/- 9.2 pmol/L; P = 0.001). In both normal brain tissue, obtained either intraoperatively or excised postmortem, and brain tumors, the concentrations of 3,3'-T2 ranged between 50-300 fmol/g. In healthy controls, 2 different forms of acute stress (sleep deprivation and delivering a lecture) significantly increased serum levels of T4 and T3, but did not affect those of 3,3'-T2 or 3,5-T2. In conclusion, our results show that, contrary to expectation, a low T3 syndrome in NTI is not always associated with low serum concentrations of 3,3'-T2. The production of 3,3'-T2 in NTI seems to be regulated in a disease-specific manner, resulting in unchanged, reduced, or elevated hormone concentrations. PMID- 9745406 TI - Prevalence of the polycystic ovary syndrome in unselected black and white women of the southeastern United States: a prospective study. AB - Estimates of the prevalence of the polycystic ovary syndrome (PCOS) in the general population have ranged from 2-20%. The vast majority of these reports have studied White populations in Europe, used limited definitions of the disorder, and/or used bias populations, such as those seeking medical care. To estimate the prevalence of this disorder in the United States and address these limitations, we prospectively determined the prevalence of PCOS in a reproductive aged population of 369 consecutive women (174 White and 195 Black; aged 18-45 yr), examined at the time of their preemployment physical. Body measures were obtained, and body hair was quantified by a modified Ferriman-Gallwey (F-G) method. All exams were initially performed by 2 trained nurses, and any subject with an F-G score above 3 was reexamined by a physician, the same for all patients. Of the 369 women, 277 (75.1%) also agreed to complete a questionnaire and have additional blood drawn. Subjects were studied regardless of current estrogen/progestin hormonal use (28.5%). PCOS was defined as 1) oligoovulation, 2) clinical hyperandrogenism (i.e. hirsutism) and/or hyperandrogenemia, and 3) exclusion of other related disorders, such as hyperprolactinemia, thyroid abnormalities, and non-classic adrenal hyperplasia. Hirsutism was defined by a F G score of 6 or more, and hyperandrogenemia was defined as a total or free testosterone, androstenedione, and/or dehydroepiandrosterone sulfate level above the 95th percentile of control values [i.e. all eumenorrheic women in the study, who had no hirsutism (F-G < or = 5) or acne and were receiving no hormonal therapy; n = 98]. Considering all 369 women studied, White and Black women had similar mean ages (29.4 +/- 7.1 and 31.1 +/- 7.8 yr, respectively), although White women had a lesser body mass than Black women (24.9 +/- 6.1 vs. 29.2 +/- 8.1 kg/m2, respectively; P < 0.001). Of these 7.6%, 4.6%, and 1.9% demonstrated a F-G score of 6 or more, 8 or 10, respectively, and there was no significant racial difference, with hirsutism prevalences of 8.0%, 2.8%, and 1.6% in Whites, and 7.1%, 6.1%, and 2.1% in Blacks, respectively. Of the 277 women consenting to a history and hormonal evaluation, 4.0% had PCOS as defined, 4.7% (6 of 129) of Whites and 3.4% (5 of 148) of Blacks. In conclusion, in our consecutive population of unselected women the prevalence of hirsutism varied from 2-8% depending on the chosen cut-off F-G score, with no significant difference between White and Black women. Using an F-G score of 6 or more as indicative of hirsutism, 3.4% of Blacks and 4.7% of Whites had PCOS as defined. These data suggest that PCOS may be one of most common reproductive endocrinological disorders of women. PMID- 9745407 TI - Menstrual abnormalities in women with Cushing's disease are correlated with hypercortisolemia rather than raised circulating androgen levels. AB - Menstrual irregularity is a common complaint at presentation in women with Cushing's syndrome, although the etiology has been little studied. We have assessed 45 female patients (median age, 32 yr; range, 16-41 yr) with newly diagnosed pituitary-dependent Cushing's syndrome. Patients were subdivided into 4 groups according to the duration of their menstrual cycle: normal cycles (NC; 26 30 days), oligomenorrhea (OL; 31-120 days), amenorrhea (AM; > 120 days), and polymenorrhea (PM; < 26 days). Blood was taken at 0900 h for measurement of LH, FSH, PRL, testosterone, androstenedione, dehydroepiandrosterone sulfate, estradiol (E2), sex hormone-binding globulin (SHBG), and ACTH; cortisol was sampled at 0900, 1800, and 2400 h. The LH and FSH responses to 100 micrograms GnRH were analyzed in 23 patients. Statistical analysis was performed using the nonparametric Mann-Whitney U and Spearman tests. Only 9 patients had NC (20%), 14 had OL (31.1%), 15 had AM (33.3%), and 4 had PM (8.8%), whereas 3 had variable cycles (6.7%). By group, AM patients had lower serum E2 levels (median, 110 pmol/L) than OL patients (225 pmol/L; P < 0.05) or NC patients (279 pmol/L; P < 0.05), and higher serum cortisol levels at 0900 h (800 vs. 602 and 580 nmol/L, respectively; P < 0.05) and 1800 h (816 vs. 557 and 523 nmol/L, respectively; P < 0.05) and higher mean values from 6 samples obtained through the day (753 vs. 491 and 459 nmol/L, respectively; P < 0.05). For the whole group of patients there was a negative correlation between serum E2 and cortisol at 0900 h (r = -0.50; P < 0.01) and 1800 h (r = -0.56; P < 0.01) and with mean cortisol (r = -0.46; P < 0.05). No significant correlation was found between any serum androgen and E2 or cortisol. The LH response to GnRH was normal in 43.5% of the patients, exaggerated in 52.1%, and decreased in 4.4%, but there were no significant differences among the menstrual groups. No differences were found in any other parameter. In summary, in our study 80% of patients with Cushing's syndrome had menstrual irregularity, and this was most closely related to serum cortisol rather than to circulating androgens. Patients with AM had higher levels of cortisol and lower levels of E2, while the GnRH response was either normal or exaggerated. Our data suggest that the menstrual irregularity in Cushing's disease appears to be the result of hypercortisolemic inhibition of gonadotropin release acting at a hypothalamic level, rather than raised circulating androgen levels. PMID- 9745408 TI - Dynamic endocrine testing and magnetic resonance imaging in the long-term follow up of childhood langerhans cell histiocytosis. AB - Children treated for Langerhans cell histiocytosis (LCH) are at risk for short and long term endocrine sequelae, but biological predictors of specific deficits are not well defined. We evaluated the frequency and progression of LCH-related endocrine deficits during long term follow-up and assessed the ability of dynamic endocrine testing to identify patients at risk for late anterior or posterior pituitary hormone dysfunction. The 17 patients (5 males and 12 females) were followed a median of 10 yr after diagnosis of single system (n = 6) or multisystem (n = 11) disease. Study evaluations, performed a median of 4.1 yr after the diagnosis, comprised pituitary hormone responses to the appropriate challenge, 7-h water deprivation test, 3% hypertonic saline infusion, and magnetic resonance imaging (MRI). The six patients with GH deficiency at the time of evaluation had a significantly lower GH response to GHRH than the other patients [median peak, 7.3 vs. 21.5 micrograms/L (P = 0.03); median area under the curve, 4.7 vs. 13.5 micrograms/L (P = 0.03)]; levels in the latter group did not differ significantly from those in 20 age- and sex-matched controls with constitutional or familial short stature. Two patients who had GH responses to GHRH of 20.6 and 23 ng/mL at 2.8 and 9.5 yr of age developed GH deficiency at 6.5 and 11.2 yr of age, respectively. The TSH response to TRH was less than 10 mU/L in three patients, two of whom later developed central hypothyroidism. ACTH and cortisol responses to CRF, and PRL responses to TRH were normal in all cases, and LH and FSH responses to GnRH were compatible with pubertal stage. Abnormalities in arginine vasopressin responses to water deprivation or hypertonic saline infusion were seen only in four patients who had preexisting diabetes insipidus (DI); one patient who later developed DI had normal findings. On standard MRI, posterior pituitary hyperintensity was absent only in the patients with DI. Pituitary stalk thickening was seen in seven patients, including three who did not have DI and had normal arginine vasopressin responses. Delayed posterior and anterior enhancement on dynamic MRI was present in two patients, both of whom later developed central hypothyroidism. Patients with single system disease had a lower 5-yr probability of LCH reactivation (41% vs. 83% for those with multisystem disease; P = 0.21) and a significantly lower risk of endocrine dysfunction (P = 0.007). In this series, dynamic evaluation of pituitary function was not a useful predictor of late endocrine sequelae, with the possible exception of the progressively decreasing TSH response to TRH. Similarly, a standard MRI was not predictive, although dynamic imaging may be informative regarding evolving pituitary hormone deficiency. PMID- 9745409 TI - Salsalate administration--a potential pharmacological model of the sick euthyroid syndrome. AB - This study examined salsalate ingestion as a model of the sequelae of acute inhibition of thyroid hormone binding to serum protein. One dose of salsalate (60 65 mg/kg) was administered to healthy volunteers. Serum salsalate concentrations peaked at 2 h (82 micrograms/mL), then declined at 8 h to 1.2 micrograms/mL. Serum total T4 (TT4) and total T3 (TT3) concentrations declined for 4 h, then recovered by 96 h, while T4 binding protein concentrations remained unchanged. TT3 was reduced to a greater extent than TT4 between 2 h and 72 h, and serum total reverse(r)T3 (TrT3) was transiently increased at 8 h. TSH concentrations fell while TT4 and TT3 fell, then recovered while TT4, TT3, and free T3, but not free T4, were still reduced. Subsequently, TSH overshot basal levels and continued to rise after 96 h while TT4, TT3, free T4, free T3, and TrT3 were all at basal levels. We postulate that an acute release of T4 and T3 from circulating transport proteins, induced by an inhibitor of binding, can result in large and rapid redistribution of T4 and T3 into tissue compartments associated with transiently reduced peripheral tissue 5'-monodeiodination and deranged TSH regulation. PMID- 9745410 TI - Is there a role for low doses of mitotane (o,p'-DDD) as adjuvant therapy in adrenocortical carcinoma? AB - Four patients suffering from adrenocortical carcinoma were treated with low doses (1.5-2.0 g) of mitotane (o,p'-DDD) for the complete follow-up time following surgery (21-68 months). Treatment with mitotane was started shortly after surgical removal of the tumor (three patients) or the tumor and multiple lung metastasis (one patient). No significant side effects or complications from the medication were noted. Two patients remain disease free after 57 and 21 months on treatment. A third patient died of an unrelated reason (varicose vein bleeding) after 68 months on mitotane without evidence of tumor recurrence or metastasis. In the fourth patient, two lung metastasis were successfully removed after 48 months of follow-up. The patient is doing well and is disease free 6 months later. Though our series is too small to draw final conclusions, we suggest that low doses of mitotane, which are well tolerated, might offer prolonged disease free survival in adrenocortical carcinoma. To be beneficial treatment has to be started early after surgical removal of the tumor and metastasis, and be continued for long periods of time. PMID- 9745411 TI - Long-acting lanreotide induces clinical and biochemical remission of acromegaly caused by disseminated growth hormone-releasing hormone-secreting carcinoid. AB - Ectopic GHRH-secreting tumors, such as carcinoid, rarely cause acromegaly. As protracted exposure to high levels of GH is associated with considerable morbidity and mortality, these patients require early and effective medical therapy to control hormonal hypersecretion. We employed a prolonged release somatostatin analog, lanreotide, to treat a patient with disseminated GHRH producing carcinoid. Before treatment, the patient had a biochemical profile characteristic of active acromegaly. Plasma GHRH levels were markedly elevated (200-fold), and urinary 5-hydroxyindolacetic acid (5-HIAA) levels were increased (4-fold). Magnetic resonance imaging revealed a large asymmetrical pituitary mass consistent with somatotroph hyperplasia. Somatostatin receptor scintigraphy revealed multiple bony and soft tissue lesions as well as striking pituitary uptake. Lanreotide (30 mg) was administered weekly by im injection for 12 weeks. Rapid and sustained symptomatic clinical improvement with diminished soft tissue swelling and hyperhidrosis was observed. GHRH levels decreased by 70%; glucose suppressed GH and insulin-like growth factor I levels were reduced by 90% and 75%, respectively, to near normal values; urinary 5-HIAA levels normalized; and the pituitary mass remained unchanged. Unfortunately, the patient died due to complications of osteogenic sarcoma. In conclusion, prolonged release lanreotide induced clinical and biochemical remission in this patient with diffusely metastatic GHRH-producing carcinoid. This long-acting drug thus offers an effective, well tolerated, and convenient medical therapy for control of hormonal hypersecretion induced by excess GHRH. PMID- 9745412 TI - Serum inhibin B as a marker of spermatogenesis. AB - Inhibin B is produced by Sertoli cells, provides negative feedback on FSH secretion, and may prove to be an important marker for the functioning of seminiferous tubules. The purpose of the present study was to examine the relationship between the spermatogenic function of the testis of subfertile men and the plasma concentrations of inhibin B and FSH. These parameters were estimated in a group of 218 subfertile men. Serum inhibin B levels were closely correlated with the serum FSH levels (r = -0.78, P < 0.001), confirming the role of inhibin B as feedback signal for FSH production. The spermatogenic function of the testis was evaluated by determining testicular volume and total sperm count. Inhibin B levels were significantly correlated with the total sperm count and testicular volume (r = 0.54 and r = 0.63, respectively; P < 0.001). Testicular biopsies were obtained in 22 of these men. Inhibin B was significantly correlated with the biopsy score (r = 0.76, P < 0.001). Receiver operating characteristic analysis revealed a diagnostic accuracy of 95% for differentiating competent from impaired spermatogenesis for inhibin B, whereas for FSH, a value of 80% was found. We conclude that inhibin B is the best available endocrine marker of spermatogenesis in subfertile men. PMID- 9745413 TI - Metabolic consequences of 5-year growth hormone (GH) therapy in children treated with GH for idiopathic short stature. Genentech Collaborative Study Group. AB - In a multicenter study the metabolic effects of 5 yr of GH therapy in children with idiopathic short stature were evaluated. Patients received 0.3 mg/kg.week recombinant human GH. Of the 121 patients who entered the study, data for 62 were analyzed at the final 5 yr point. Routine laboratory determinations were available for all 62 subjects at the 5 yr point. Special laboratory determinations, such as postprandial glucose and insulin, were available for only a subset of patients. Mean insulin-like growth factor I levels rose to 283 +/- 101 micrograms/L, within the normal range using age-appropriate reference standards. T4, cholesterol, triglycerides, blood chemistries, and blood pressure showed no significant changes during the 5-yr period. Mean baseline and 2-h postprandial glucose levels remained unchanged. Both fasting and postprandial insulin levels rose substantively from low normal levels to the normal range (median, 4.9-43 mU/L). Mean hemoglobin A1c levels remained within the normal range throughout the study. In summary, careful monitoring has not revealed any currently discernible metabolic side-effects of clinical significance after GH therapy in this 5-yr study of children with idiopathic short stature. PMID- 9745414 TI - Effects of intensive chemotherapy on bone and collagen turnover and the growth hormone axis in children with acute lymphoblastic leukemia. AB - To investigate the effects of disease and intensive chemotherapy on bone turnover and growth in children with acute lymphoblastic leukemia (ALL), a longitudinal prospective study was carried out in 22 children, aged 1.2-13.5 yr, enrolled in the Medical Research Council-funded randomized trial of childhood ALL treatment in the UK. We measured lower leg length and markers of bone formation [bone alkaline phosphatase (ALP) and procollagen type I C-terminal propeptide (PICP)], bone resorption [pyridinoline, deoxypyridinoline, and carboxyl-terminal telopeptide of type I collagen (ICTP)], soft tissue turnover [procollagen type III N-terminal propeptide (P3NP)], and the GH axis [IGF-I, IGF-binding protein-3 (IGFBP-3), IGFBP-2, and urinary GH] at 1- to 4-week intervals from diagnosis to week 27 of treatment. In addition, GH-binding protein was measured at diagnosis. At diagnosis, mean SD scores were: bone ALP, -1.84; PICP -1.77; pyridinoline, 1.42; deoxypyridinoline, -1.66; ICTP, -0.42; P3NP, +1.45; GH, +24.4; IGF-I, 1.70; IGFBP-3, -0.88; IGFBP-2, +2.42; and GH-binding protein, -0.69. Bone ALP, PICP, and IGFBP-3 were all correlated (P < or = 0.03). During induction and intensification, there was shrinkage of the lower leg, with decreases in PICP, pyridinoline, ICTP, and P3NP (P < 0.05), whereas IGF-I and IGFBP-3 increased (P < 0.05). After prednisolone was discontinued, bone ALP and collagen markers increased markedly (P < 0.01), but there was no significant change in IGF-I and IGFBP-3. In 12 children who received high dose i.v. methotrexate, postglucocorticoid increases in bone ALP and PICP were less, whereas those in ICTP and P3NP were greater, compared to levels in children who did not receive methotrexate (P < 0.05). We conclude that ALL itself caused GH resistance and low bone turnover. During early intensive chemotherapy, further suppression of osteoblast proliferation and osteoclast activity occurred, not mediated through the systemic GH axis, probably by the direct action of prednisolone on bone. The postglucocorticoid increase in bone turnover was also independent of the GH axis and was modulated by high dose i.v. methotrexate, which depressed osteoblast recovery and enhanced osteoclast activity. PMID- 9745415 TI - Increased diurnal plasma concentrations of dehydroepiandrosterone in depressed patients. AB - Activation of the hypothalamus-pituitary-adrenocortical system is a biological core symptom of depression. Although the regulation of cortisol secretion is well studied in this condition, there is no information about the diurnal activity of dehydroepiandrosterone (DHEA) secretion. Therefore, we studied 24-h DHEA plasma concentrations (every 30 min) in severely depressed patients (n = 26) and healthy controls (n = 33). We found depression to significantly increase diurnal minimal and mean DHEA plasma concentrations, whereas there was no effect on the diurnal maximal plasma concentration and the diurnal amplitude of DHEA. In particular, we found a parallel increase in mean DHEA (5.8 +/- 3.6 vs. 3.4 +/- 1.9 nmol/L; P < 0.003), cortisol (286 +/- 65 vs. 184 +/- 29 nmol/L; P < 0.0001) and ACTH (7.14 +/ 2.06 vs. 5.72 +/- 1.36 pmol/L; P < 0.002) plasma concentrations. The novel finding of parallel increases in diurnal DHEA and cortisol plasma concentrations in depressed patients has important implications for the regulation of the hypothalamus-pituitary-adrenocortical system in conditions of chronic stress and for the rationale of DHEA treatment in depressed patients. PMID- 9745416 TI - Cushing's syndrome due to a gastric inhibitory polypeptide-dependent adrenal adenoma: insights into hormonal control of adrenocortical tumorigenesis. AB - We studied a patient with food-induced, ACTH-independent, Cushing's syndrome and a unilateral adrenocortical adenoma. In vivo cortisol secretion was stimulated by mixed, glucidic, lipidic, or proteic meals. Plasma ACTH levels were undetectable, but iv injection of ACTH stimulated cortisol secretion. Unilateral adrenalectomy was followed by hypocortisolism with loss of steroidogenic responses to both food and ACTH. In vitro, cortisol secretion by isolated tumor cells was stimulated by the gut hormone gastric inhibitory polypeptide (GIP) and ACTH, but not by another gut hormone, glucagon-like peptide-1 (GLP-1). Both peptides stimulated the production of cAMP but not of inositol 1,4,5-trisphosphate. In quiescent cells, GIP and ACTH stimulated [3H]thymidine incorporation and p42-p44 mitogen-activated protein kinase activity. GIP receptor messenger ribonucleic acid (RNA), assessed by RT-PCR, was highly expressed in the tumor, whereas it was undetectable in the adjacent hypotrophic adrenal tissue, in two adrenal tumors responsible for food independent Cushing's syndrome, and in two hyperplastic adrenals associated with ACTH hypersecretion. In situ hybridization demonstrated that expression of GIP receptor RNA was confined to the adrenocortical tumor cells. Low levels of ACTH receptor messenger RNA were also detectable in the tumor. We conclude that abnormal expression of the GIP receptor allows adrenocortical cells to respond to food intake with an increase in cAMP that may participate in the stimulation of both cortisol secretion and proliferation of the tumor cells. PMID- 9745417 TI - Normal bone mass in bulimic women. AB - The aim of this study was to examine the relationship among exercise, menstrual function, and bone mineral density (BMD) in different groups of age-matched patients with eating disorders. Dieting and eating disorder history, physical activity history, and menstrual history were assessed by clinical interview in 43 bulimic and 13 anorectic young women as well as in 17 healthy control subjects (18-29 yr). BMD was assessed by dual x-ray absorptiometry. All the anorectics but only 30% of the bulimics exercised regularly from the onset of their eating disorder (P < 0.01), mainly using aerobic dancing and running. All of the anorectics had been amenorrheic since the start of their symptoms, and 68% of the bulimics had a history of menstrual dysfunction. Within the exercise subgroups of bulimic patients, there was no significant relationship between BMD and current or previous menstrual function. Anorectic patients had lower BMD than bulimics and controls in all skeletal regions studied (P < 0.01). Bulimic patients who had exercised regularly during their illness had higher total body BMD than bulimics classified as sedentary (P < 0.01). Bulimics who had exercised regularly or intermittently since the onset of their eating disorder had higher BMD than sedentary bulimics in the lumbar vertebrae, femoral neck, and legs (P < 0.05). It appears that weight-bearing exercise can prevent or attenuate bone loss at specific skeletal sites in normal weight bulimic patients, but not in anorectics. PMID- 9745418 TI - Measurement of volumetric bone mineral density accurately determines degree of lumbar undermineralization in children with growth hormone deficiency. AB - The effect of anthropometric variables and bone size on bone mineral density (BMD) was examined in 22 children with GH deficiency (GHD) aged 6.1-8.0 yr at diagnosis and in 40 sex- and chronological age-matched controls. In all patients and controls, bone mineral content (BMC), BMDarea and BMD corrected for the apparent bone volume (BMDvolume) were measured by dual-energy x-ray absorptiometry in the lumbar spine at L2-L4 level. In patients, BMDarea was corrected for body height (BMDheight), body mass index (BMDBMI), and bone age (BMDBA). Patients showed significantly reduced (P < 0.0001) BMC (males 11.55 +/- 0.71 g, females 10.13 +/- 1.48 g) and BMDarea (males 0.502 +/- 0.033 g/cm2, females 0.515 +/- 0.034 g/cm2) compared with controls (BMC: males 18.09 +/- 1.23 g, females 15.58 +/- 1.87 g; BMDarea: males 0.689 +/- 0.065 g/cm2, females 0.685 +/- 0.059 g/cm2). In patients, BMDheight (males 0.537 +/- 0.031 g/cm2, females 0.548 +/- 0.032 g/cm2) and BMDBMI (males 0.641 +/- 0.028 g/cm2, females 0.624 +/- 0.035 g/cm2) remained significantly lower (P < 0.02 to P < 0.0001) than BMDarea of controls. BMDBA of patients was significantly reduced (-1.49 +/- 0.51 Z score, P < 0.0001) in comparison with bone age-matched controls (n = 35). BMDvolume was significantly lower (P < 0.01 to P < 0.0005) in patients (males 0.268 +/- 0.006 g/cm3, females 0.276 +/- 0.010 g/cm3) compared with chronological age-matched controls (males 0.283 +/- 0.013 g/cm3, females 0.293 +/- 0.017 g/cm3). Mean bone volume of patients was affected to a greater extent than bone area (-2.36 +/- 0.49 Z score and -1.56 +/- 0.70 Z score, respectively). Bone area/bone volume ratio was significantly higher in patients than in chronological age-matched controls (0.53 +/- 0.02 and 0.42 +/- 0.08, P < 0.0001, respectively). Chronological age, body height, BMI, and bone age correlated significantly with BMDarea (r2 = 0.389-0.450, P < 0.002 to P < 0.001) but not with BMDvolume (P = not significant). The results show that anthropometric variables and bone size affect lumbar BMC and BMDarea in children with GHD. Reduced lumbar BMDvolume indicates that apparent true bone density is decreased in children with GHD, suggesting a role of GH in bone mineralization. PMID- 9745419 TI - Effects of testosterone replacement with a nongenital, transdermal system, Androderm, in human immunodeficiency virus-infected men with low testosterone levels. AB - Although weight loss associated with human immunodeficiency virus (HIV) infection is multifactorial in its pathogenesis, it has been speculated that hypogonadism, a common occurrence in HIV disease, contributes to depletion of lean tissue and muscle dysfunction. We, therefore, examined the effects of testosterone replacement by means of Androderm, a permeation-enhanced, nongenital transdermal system, on lean body mass, body weight, muscle strength, health-related quality of life, and HIV-disease markers. We randomly assigned 41 HIV-infected, ambulatory men, 18-60 yr of age, with serum testosterone levels below 400 ng/dL, to 1 of 2 treatment groups: group I, two placebo patches (n = 21); or group II, two testosterone patches designed to release 5 mg testosterone over 24 h. Eighteen men in the placebo group and 14 men in the testosterone group completed the 12-week treatment. Serum total and free testosterone and dihydrotestosterone levels increased, and LH and FSH levels decreased in the testosterone-treated, but not in the placebo-treated, men. Lean body mass and fat-free mass, measured by dual energy x-ray absorptiometry, increased significantly in men receiving testosterone patches [change in lean body mass, +1.345 +/- 0.533 kg (P = 0.02 compared to no change); change in fat-free mass, +1.364 +/- 0.525 kg (P = 0.02 compared to no change)], but did not change in the placebo group [change in lean body mass, 0.189 +/- 0.470 kg (P = NS compared to no change); change in fat-free mass, 0.186 +/- 0.470 kg (P = NS compared to no change)]. However, there was no significant difference between the 2 treatment groups in the change in lean body mass. The change in lean body mass during treatment was moderately correlated with the increment in serum testosterone levels (r = 0.41; P = 0.02). The testosterone-treated men experienced a greater decrease in fat mass than those receiving placebo patches (P = 0.04). There was no significant change in body weight in either treatment group. Changes in overall quality of life scores did not correlate with testosterone treatment; however, in the subcategory of role limitation due to emotional problems, the men in the testosterone group improved an average of 43 points of a 0-100 possible score, whereas those in the placebo group did not change. Red cell count increased in the testosterone group (change in red cell count, +0.1 +/- 0.1 10(12)/L) but decreased in the placebo group (change in red cell count, -0.2 +/- 0.1 10(12)/L). CD4+ and CD8+ T cell counts and plasma HIV copy number did not significantly change during treatment. Serum prostate-specific antigen and plasma lipid levels did not change in either treatment group. Testosterone replacement in HIV-infected men with low testosterone levels is safe and is associated with a 1.35-kg gain in lean body mass, a significantly greater reduction in fat mass than that achieved with placebo treatment, an increased red cell count, and an improvement in role limitation due to emotional problems. Further studies are needed to assess whether testosterone supplementation can produce clinically meaningful changes in muscle function and disease outcome in HIV-infected men. PMID- 9745421 TI - Troglitazone monotherapy improves glycemic control in patients with type 2 diabetes mellitus: a randomized, controlled study. The Troglitazone Study Group. AB - To assess the effects of troglitazone monotherapy on glycemic control in patients with type 2 diabetes mellitus, we carried out a 6-month, randomized, double blind, placebo-controlled study in 24 hospital and outpatient clinics in the United States and Canada. Troglitazone 100, 200, 400, or 600 mg or placebo once daily with breakfast was administered to 402 patients with type 2 diabetes with fasting serum glucose (FSG) > 140 mg/dL, glycosylated hemoglobin (HbA1c) > 6.5%, and fasting C-peptide > or = 1.5 ng/mL. Prior oral hypoglycemic therapy was withdrawn in patients who received it before the study. FSG, HbA1c, C-peptide, and serum insulin were evaluated at baseline and the end of the study. Analysis was performed on two subsets of patients based on prestudy therapy: Patients treated with diet and exercise only before the study (22% of patients), and those who had been receiving sulfonylurea therapy (78% of patients). Patients treated with 400 and 600 mg troglitazone had significant decreases from baseline in mean FSG and HbA1c at month 6 compared with placebo-treated patients (FSG: -51 and -60 mg/dL, respectively; HbA1c: -0.7 and -1.1%, respectively). In the diet-only subset, 600 mg troglitazone therapy resulted in a significant (P < 0.05) reduction in HbA1c (-1.35%) and a significant reduction in FSG (-42 mg/dL) compared with placebo. Patients previously treated with sulfonylurea therapy had significant (P < 0.05) decreases in mean FSG with 200-600 mg troglitazone therapy compared with placebo (-48, -61, and -66 mg/dL, respectively). Significant (P < 0.05) decreases in mean HbA1c occurred with 400 and 600 mg troglitazone therapy at month 6 (-0.8 and -1.2%, respectively) compared with placebo in this same subset. Significant (P < 0.05) decreases in triglycerides and free fatty acids occurred with troglitazone 400 and 600 mg, and increased high-density lipoprotein occurred with 600 mg troglitazone. We conclude that troglitazone monotherapy significantly improves HbA1c and fasting serum glucose, while lowering insulin and C-peptide in patients with type 2 diabetes. Troglitazone 600 mg monotherapy is efficacious for patients who are newly diagnosed and have never received pharmacological intervention for diabetes. PMID- 9745420 TI - Etiological diagnosis of primary adrenal insufficiency using an original flowchart of immune and biochemical markers. AB - Approximately 70-80% of cases of primary adrenal insufficiency are classified as idiopathic. An effective protocol for the etiological diagnosis of primary adrenal insufficiency is needed to ensure correct patient management. With the aim of developing an algorithm for the etiological diagnosis of primary adrenal insufficiency, we studied 56 Italian patients with nonsurgical primary adrenal insufficiency and 24 French patients with X-linked adrenoleukodystrophy (ALD) for serum levels of adrenal cortex, steroid-21-hydroxylase (21OHAb), islet cell (ICA), glutamate decarboxylase (GAD65Ab), IA2/ICA512 (ICA512Ab), thyroid peroxidase (TPOAb) autoantibodies, and plasmatic concentrations of very long chain fatty acids (VLCFA). High levels of 21OH and adrenal cortex antibodies were found in 35/42 (83%) and 17/42 (40%) Italian patients with idiopathic adrenal insufficiency, respectively. Levels of adrenal autoantibodies correlated inversely with disease duration (P < 0.0001). Elevated VLCFA were found in 4/42 (10%) idiopathic patients. A total of 34/35 (97%) idiopathic patients with a disease duration of less than 20 yr was positive for either 21OHSAb or elevated levels of VLCFA. None of 14 patients with posttuberculosis adrenal insufficiency had elevated levels of either adrenal antibodies or VLCFA. ICA, GAD65Ab, ICA512Ab, and TPOAb were found in 6/56 (11%), 8/56 (14%), 4/56 (7%), and 23/56 (41%) patients, respectively. None of 24 French ALD patients with adrenal insufficiency was positive for organ-specific autoantibodies. The measuring of 21OH antibodies and plasma VLCFA levels enabled a correct diagnosis of autoimmune (89%) and ALD (8%) in 97% of patients with idiopathic primary adrenal insufficiency of less than 20 yr of duration. The results of our study have important therapeutic and prognostic implications. PMID- 9745423 TI - Effects of physiological growth hormone (GH) therapy on cognition and quality of life in patients with adult-onset GH deficiency. AB - GH replacement of adults with acquired GH deficiency (GHD) results in body composition changes including increases in lean mass and bone mineral density. However, the effects of long-term GH therapy on cognitive function are largely unknown, and there are conflicting data regarding quality of life. We performed a randomized, double-blind, placebo-controlled study of GH replacement in adults with GHD and measured cognition and sense of well-being using standardized psychometric tests before and after therapy. Forty men (median age 51 yr, range 24-64 yr) with a history of pituitary disease were randomized to GH therapy (starting dose, 10 +/- 0.3 micrograms/kg per day: mean treatment dose, 4 +/- 2 micrograms/kg per day) vs. placebo for 18 months, and GH doses were adjusted according to serum insulin growth factor-I levels. At baseline, the patients displayed a full-scale intelligence quotient (IQ) score nearly 1 SD above the normal mean. Mean scores on all cognitive tests fell within normal limits, and on many tests, fell above the mean. On tests of verbal learning and delayed visual memory, mean test scores fell below the mean (although within normal limits), suggestive of a relative compromise in the area of memory performance. Following 18 months of GH replacement therapy, there were no significant changes in cognitive function or quality of life. We conclude that acquired GHD in adult men is not associated with significant alterations in cognitive function as assessed by standardized tests, and chronic low-dose GH replacement therapy does not result in significant beneficial effects on cognitive function or quality of life. Although previous studies have suggested that GH replacement in adults with acquired GHD may improve quality of life, our data do not support the use of physiological GH replacement in GHD men for this indication. PMID- 9745422 TI - Acute cardiovascular effects of insulin-like growth factor I in patients with chronic heart failure. AB - Insulin-like growth factor I (IGF-I) enhances myofibrillar development in cardiomyocytes of rats in culture and in vivo. In addition, IGF-I has vasodilatory effects and improves cardiac function in healthy volunteers. This study was conducted to evaluate the acute hemodynamic effects of IGF-I in patients with chronic heart failure Eight patients with chronic heart failure were randomized to receive recombinant human IGF-I (60 micrograms/kg) or placebo, i.v., over 4 h in a cross-over, double blind study on 2 consecutive days. Electrocardiogram as well as systemic hemodynamics were continuously monitored over 7 h by flow-guided thermodilution and radial artery catheters. IGF-I was well tolerated by all patients, and no pathological changes on electrocardiogram were recorded. Compared with placebo, IGF-I increased the cardiac index by 27 +/- 3.7% (+/- SE; P < 0.0005) and the stroke volume index by 21 +/- 5.6% (P < 0.05), and decreased systemic vascular resistance by 28 +/- 4.4% (P < 0.0002), right atrial pressure by 33 +/- 9.0% (P < 0.003), and pulmonary artery wedge pressure by 25 +/- 6.1% (P < 0.03). Mean systemic and pulmonary artery pressure as well as heart rate and pulmonary vascular resistance were not significantly influenced by IGF-I treatment. Insulin and C peptide levels were decreased by IGF-I, whereas glucose and electrolyte levels remained unchanged. Urinary levels of norepinephrine decreased significantly (P < 0.05) during IGF-I infusion. Thus, acute administration of IGF-I in patients with chronic heart failure is safe and improves cardiac performance by afterload reduction and possibly by positive inotropic effects. Further investigations to establish whether the observed acute effects of IGF-I are maintained during chronic therapy appear to be warranted. PMID- 9745424 TI - Nocturnal blood pressure elevation in patients with type 1 diabetes receiving intensive insulin therapy compared with that in patients receiving conventional insulin therapy. AB - Studies have shown that type 1 diabetic patients may suffer from nocturnal elevation in blood pressure and that this elevation may be related to hyperinsulinemia. In this study we tested the hypothesis that tight type 1 diabetes control, which is usually accompanied by hyperinsulinemia and subclinical nocturnal hypoglycemia, may result in a higher rise in nocturnal blood pressure compared with conventional type 1 diabetes control. Eighteen patients treated with intensive insulin therapy (multiple daily injections; IIT) were compared with 18 patients treated with conventional insulin regimens (twice daily injections of regular and intermediate acting insulin; CIT). Both groups were matched for age, sex, duration of diabetes, body weight, body mass index, baseline daytime blood pressure, and microalbuminuria levels. Hemoglobin A1c was lower in the IIT group compared with that in the CIT group (8.1 +/- 1.2% vs. 11.0 +/- 3.2%; P < 0.01). The amount of insulin/body weight (units per kg) was higher in the IIT group than that in the CIT group (1.0 +/- 0.2 vs. 0.7 +/- 0.2 U/kg; P < 0.05). In all patients, a 24-h ambulatory blood pressure was recorded. The nocturnal diastolic blood pressure was higher in the IIT group (66 +/- 9 mm Hg) than in the CIT group (55 +/- 4 mm Hg; P < 0.01). The nocturnal decline in both systolic and diastolic blood pressure was lower in the IIT group (7 +/- 5 and 6 +/- 4 mm Hg, respectively) compared with that in the CIT group (13 +/- 6 and 16 +/- 6 mm Hg, respectively; P < 0.01). The nocturnal heart rate was higher in IIT group than in the CIT group (81 +/- 12 vs. 67 +/- 9/min; P < 0.05). These findings show that the intensive insulin therapy regimen may have a more deleterious effect than the conventional insulin therapy regimen on the nocturnal blood pressure of patients with type 1 diabetes. PMID- 9745425 TI - Are autoimmune thyroid dysfunction and depression related? AB - The objective of this study was to examine the relationship between autoimmune thyroid disease and depression in perimenopausal women. Thyroid function [TSH, free T4, and thyroid peroxidase antibodies (TPO-Ab)] and depression (using the Edinburgh Depression Scale) were assessed cross-sectionally together with other determinants of depression. The subjects were 583 randomly selected perimenopausal women (aged 47-54 yr) from a community cohort of 6846 women. The main outcome measures were the occurrence of thyroid dysfunction (abnormal free T4 and/or TSH or elevated levels of TPO-Ab) and the concomitant presence of depression according to the Edinburgh Depression Scale. Neither biochemical thyroid dysfunction nor menopausal status was related to depression. Apart from several psycho-social determinants (the occurrence of a major life event, a previous episode of depression, or financial problems), an elevated level of TPO Ab (> or = 100 U/mL) was significantly associated with depression (odds ratio, 3.0, 95% confidence interval, 1.3-6.8). We conclude that women with elevated TPO Ab levels are especially vulnerable to depression, whereas postmenopausal status does not increase the risk of depression. PMID- 9745426 TI - Effects of estrogen on nonverbal processing speed and motor function in girls with Turner's syndrome. AB - The Turner syndrome (TS) phenotype is characterized by a specific neurocognitive profile of normal verbal skills, impaired visual-spatial and/or visual-perceptual abilities, and difficulty with motor function. In the current study, we investigated motor function and nonverbal processing speed in estrogen- and placebo-treated girls (aged 10-12 years) with TS and in age-matched female controls. The goal of this study was to examine whether estrogen replacement therapy would reverse deficits in motor function and in nonverbal processing speed, a measure of the time required to perform certain disparate nonverbal tasks, in adolescent girls with TS. Children received either estrogen (ethinyl estradiol, 12.5-50 ng/kg.day), or placebo for durations of 1-7 yr (mean, 4.0 +/- 2.1 yr) in this randomized, double blind study. Cognitive and motor tasks administered included the Wechsler Intelligence Scale for Children-Revised; nonspatial, repetitive motor tasks (tapping and three tasks from the Paness); and spatially mediated motor tasks [nongrooved pegboard (Lafayette), pursuit rotor, visual-motor integration, and money street map]. Questionnaires administered included the Self-Concept Scale. The major result of this study was the positive estrogen treatment effect on nonverbal processing speed and speeded motor performance in 12-yr-old TS girls. That motor performance would be slower in estrogen-deficient TS females is consistent with previous studies of the influence of estrogen on motor function. Estrogen replacement is thus the most likely explanation for the improved motor speed and nonverbal processing time in the estrogen-treated TS girls compared to that in the placebo-treated TS girls. Whether these findings will influence the psychoeducational outcome or quality of life of females with TS is not yet known. PMID- 9745427 TI - Novel mutations in aquaporin-2 gene in female siblings with nephrogenic diabetes insipidus: evidence of disrupted water channel function. AB - Novel mutations of the aquaporin-2 (AQP2) gene have been detected in Japanese female siblings with autosomal-recessive nephrogenic diabetes insipidus. The patients were compound heterozygote for point mutations at nucleotide position 374 (C374T) and at position 523 (G523A) in exon 2 of the AQP2 gene, resulting in substitution of methionine for threonine at codon 125 (T125M) and arginine for glycine at codon 175 (G175R). The water permeability (Pf) of oocytes injected with wild-type complementary RNA increased 9.0-fold compared with the Pf of water injected oocytes, whereas the increases in the Pf of oocytes injected with T125M and G175R complementary RNA were only 1.7-fold and 1.5-fold, respectively. Immunoblot and immunocytochemistry indicated that the plasma membrane expressions of T125M and G175R AQP2 proteins were comparable to that of the wild-type, suggesting that although neither the T125M nor G175R mutation had a significant effect on plasma membrane expression, they both distorted the structure and function of the aqueous pore of AQP2. These results provide evidence that the nephrogenic diabetes insipidus in patients with T125M and G175R mutations is attributable not to the misrouting of AQP2, but to the disrupted water channel function. PMID- 9745428 TI - The MEN-1 gene is rarely down-regulated in pituitary adenomas. AB - The gene for multiple endocrine neoplasia type 1 (MEN-1) has recently been cloned and encodes a putative tumor suppressor protein named menin. We have previously reported inactivating MEN-1 gene mutations associated with loss of heterozygosity (LOH) of the normal allele in tumors of patients with MEN-1 and in some sporadic pituitary tumors. These genetic alterations, however, are noted in no more than 10% of sporadic adenomas. To investigate whether other mechanisms may result in down-regulation of menin gene expression in pituitary adenomas, we examined menin gene expression by semiquantitative RT-PCR in 60 sporadic pituitary adenomas. Ribonucleic acid (RNA) was extracted from surgically resected, morphologically characterized tumors. Primers were designed to amplify a 257-bp fragment spanning exons 4-6 of the MEN-1 gene. A product of the predicted size was amplified from normal pituitary samples as well as from adenomas. Competitive PCR was performed with the housekeeping gene PGK-1 to quantitate menin gene expression. A comparable ratio of menin/PGK-1 messenger RNA was identified in all but three samples; in two tumors with LOH, menin expression was weak, and in one tumor, menin messenger RNA was undetectable, associated with LOH and mutation of the other allele. Reduced expression of menin in some sporadic adenomas is consistent with a putative tumor suppressor role for this gene product. However, lack of menin down-regulation in the majority of these tumors, which exhibit LOH at 11q13 in up to 20% of cases, provides compelling evidence for an additional tumor suppressor gene at this locus, which is more commonly involved in the pathogenesis of pituitary neoplasms. PMID- 9745429 TI - Recommendations for nomenclature of the insulin-like growth factor binding protein superfamily. PMID- 9745430 TI - Familial hyperaldosteronism type II: description of a large kindred and exclusion of the aldosterone synthase (CYP11B2) gene. AB - Familial hyperaldosteronism type II (FH-II) is characterized by autosomal dominant inheritance and hypersecretion of aldosterone due to adrenocortical hyperplasia or an aldosterone-producing adenoma; unlike FH type I (FH-I), hyperaldosteronism in FH-II is not suppressible by dexamethasone. Of a total of 17 FH-II families with 44 affected members, we studied a large kindred with 7 affected members that was informative for linkage analysis. Family members were screened with the aldosterone/PRA ratio test; patients with aldosterone/PRA ratio greater than 25 underwent fludrocortisone/salt suppression testing for confirmation of autonomous aldosterone secretion. Postural testing, adrenal gland imaging, and adrenal venous sampling were also performed. Individuals affected by FH-II demonstrated lack of suppression of plasma A levels after 4 days of dexamethasone treatment (0.5 mg every 6 h). All patients had negative genetic testing for the defect associated with FH-I, the CYP11B1/CYP11B2 hybrid gene. Genetic linkage was then examined between FH-II and aldosterone synthase (the CYP11B2 gene) on chromosome 8q. A polyadenylase repeat within the 5'-region of the CYP11B2 gene and 9 other markers covering an approximately 80-centimorgan area on chromosome 8q21-8qtel were genotyped and analyzed for linkage. Two-point logarithm of odds scores were negative and ranged from -12.6 for the CYP11B2 polymorphic marker to -0.98 for the D8S527 marker at a recombination distance (theta) of 0. Multipoint logarithm of odds score analysis confirmed the exclusion of the chromosome 8q21-8qtel area as a region harboring the candidate gene for FH II in this family. We conclude that FH-II shares autosomal dominant inheritance and hyperaldosteronism with FH-I, but, as demonstrated by the large kindred investigated in this report, it is clinically and genetically distinct. Linkage analysis demonstrated that the CYP11B2 gene is not responsible for FH-II in this family; furthermore, chromosome 8q21-8qtel most likely does not harbor the genetic defect in this kindred. PMID- 9745431 TI - Androgen response to hypothalamic-pituitary-adrenal stimulation with naloxone in women with myotonic muscular dystrophy. AB - Myotonic muscular dystrophy (MMD) is a disease of autosomal dominant inheritance characterized by multisystem disease, including myotonia, muscle-wasting and weakness of all muscular tissues, and endocrine abnormalities attributed to a genetic abnormality causing a defective cAMP-dependent kinase. We have previously reported that MMD patients demonstrate ACTH hypersecretion after endogenous CRH release stimulated by naloxone administration while manifesting a normal cortisol (F) response. Additionally, others have reported a reduced adrenal androgen (AA) response to exogenous ACTH administration in MMD patients. As ACTH stimulates the secretion of both AAs and F, it is possible that the discordant relationship of these hormones in MMD patients results from a defect of adrenocortical ACTH receptor function or postreceptor signaling or subsequent biochemical events. Furthermore, the molecular abnormality seen in MMD patients may suggest that the mechanism underlying the frequently observed discordances in the secretion of glucocorticoids and AAs (e.g. adrenarche, surgical trauma, severe burns, or intermittent glucocorticoid administration) are explainable solely via an alteration in the function of the ACTH receptor or postreceptor signaling. To ascertain whether the responses of F and AAs to endogenous ACTH diverged in this disorder, we prospectively studied the responses of these hormones to naloxone stimulated CRH release in nine premenopausal women with MMD and seven healthy age and weight-matched control women. After naloxone infusion (125 micrograms/kg, i.v.), blood sampling was performed at baseline (i.e. -5 min) and at 30 and 60 min. In addition to the absolute hormone level at each time, we calculated the net increment (i.e. change) at 30 and 60 min and the area under the curve (AUC) for F, ACTH, dehydroepiandrosterone (DHA), and androstenedione (A4). Consistent with our previous study, MMD patients demonstrated higher ACTH levels at all sampling times except [minud]5 min. AUC analysis revealed the ACTHAUC values were significantly higher in MMD than in control women (457 +/- 346 vs. 157 +/- 123 pmol/min.L; P < 0.03), whereas the FAUC response did not differ between MMD and controls (13860 +/- 3473 vs. 13375 +/- 3465 nmol/min.L; P > 0.5). Despite the greater ACTH secretion, the baseline circulating dehydroepiandrosterone sulfate levels were significantly lower in MMD compared with control women (18 +/- 23 vs. 61 +/- 23 mumol/L; P < 0.002). The serum concentrations of A4 at baseline, 30 min, and 60 min and DHA levels at 30 and 60 min were also significantly lower in MMD vs. control women. Additionally, the A4AUC and DHAAUC values were significantly lower in MMD patients than in controls. Furthermore, the net response of DHA at 60 min to the endogenous ACTH increase was also reduced in MMD patients compared with that in control subjects (2.3 +/- 2.1 vs. 5.6 +/- 2.6 nmol/L; P < 0.02). In conclusion, in addition to ACTH hypersecretion to CRH mediated stimuli, these data suggest that MMD patients have a defect in the adrenocortical response to ACTH, reflected in normal F and reduced DHA and A4 secretion. Whether this defect is inherent to the disease or simply reflects adaptive changes to chronic disease remains to be demonstrated. However, it is possible that further studies of the response of MMD patients to ACTH may reveal a mechanism that explains the frequently observed dichotomy in the secretion of glucocorticoids and AAs. PMID- 9745433 TI - Different central and peripheral responses to leptin in rhesus monkeys: brain transport may be limited. AB - The purpose of this experiment was to determine the effect of leptin administration on food intake and energy expenditure in rhesus monkeys. Four adult male rhesus monkeys, cannulated in the left lateral cerebral ventricle, were used for all phases of this experiment. Food intake was measured following intracerebroventricular injections of vehicle or three doses (500 ng, 2 micrograms, and 22 micrograms) leptin. Leptin administration resulted in a dose dependent decrease in food intake (P < 0.05), with food intake decreased by an average of 54% at 22 micrograms leptin. Energy expenditure was also measured at two intracerebroventricular doses of leptin. Energy expenditure was not different (P > 0.10) between placebo and leptin injections at either dose. Food intake was also measured following i.v. injection of 3 mg leptin. In this case, leptin did not alter (P > 0.10) food intake, despite increasing serum leptin levels by as much as 100-fold. These results suggest that leptin is a potent inhibitor of food intake in rhesus monkeys, but this effect requires elevation of leptin concentrations in the cerebrospinal fluid or critical brain sites. The transport system for movement of leptin across the blood-brain barrier may limit the influence of circulating leptin on food intake in monkeys. PMID- 9745432 TI - Augmented placental production of leptin in preeclampsia: possible involvement of placental hypoxia. AB - Preeclampsia (PE) is a hypertensive disorder, which develops in late pregnancy and is usually associated with placental hypoxia and dysfunction. We have recently demonstrated that leptin is a novel placenta-derived hormone in humans and suggested its significance in human pregnancy (see Ref. 19). To explore the changes in the leptin production in placenta in PE, we measured the plasma leptin level and placental leptin messenger RNA expression in pregnant women with PE. Plasma leptin levels in preeclamptic women were elevated significantly, compared with gestational age- and body mass index-matched normal pregnant women (P < 0.0001). Plasma leptin levels in the severe PE group were significantly higher than those in the mild PE group (P < 0.0001). Plasma leptin levels in preeclamptic women were reduced, soon after the placental delivery, to those expected for their body mass indices. Northern blot analysis revealed that leptin messenger RNA levels are increased in the placentas from preeclamptic women, compared with normal pregnant women. Leptin secretion was increased significantly in a human trophoblastic cell line (BeWo cells) cultured under hypoxic conditions (5% O2), compared with those cultured under standard conditions (20% O2; P < 0.01). The present study demonstrated that placental production of leptin is augmented in severe PE, probably because of placental hypoxia, thereby suggesting the possible significance of leptin as a marker of placental hypoxia in severe PE. PMID- 9745434 TI - Molecular characterization of 5 alpha-reductase type 2 deficiency and fertility in a Swedish family. AB - The molecular background of 5 alpha-reductase type 2 deficiency was investigated in a Swedish family with no known consanguinity and in which the affected males were fertile. The three male siblings were born with ambiguous external genitalia, and the diagnosis of 5 alpha-reductase deficiency was established at the ages of 16, 14, and 10 yr, respectively. All three siblings underwent surgery for hypospadias repair. At least two of the brothers are demonstrably fertile. Molecular analysis showed the three brothers to be compound heterozygotes, carrying two different mutations in exon 4 of the 5 alpha-reductase type 2 gene. The two mutations (G196S and H231R) have been described previously and reported to give rise to partially functioning enzymes, which may explain the milder phenotype and perhaps the fertility in the preset three patients. PMID- 9745435 TI - Identification of new sequence variants in the leptin gene. AB - The leptin gene (LEP) has been linked to extreme obesity. However, no common obesity-related gene variants have been found to exist in the LEP. The present study was designed to investigate the LEP for variants by screening both the putative promoter and the coding region of this gene in obese Finnish subjects (n = 200; body mass index, > 27 kg/m2). PCR-amplified DNA samples were subjected to single strand conformation analysis. A G144A substitution in codon 48 and a G328A substitution in codon 110 were identified in two obese subjects, both of whom had very low serum leptin levels. A rare silent C538T polymorphism was detected 33 bp downstream of the translation stop codon (TGA). A common polymorphism A19G was identified in the untranslated exon 1. This polymorphism was not associated with traits of obesity; in agreement, the allele frequencies were similar between 64 normal weight and 141 obese Finns. In summary, this study failed to find a common gene variant in the LEP associated with obesity, but introduces 2 rare mutations associated with very low serum leptin concentrations in 2 obese subjects. PMID- 9745436 TI - Inverse correlation between serum testosterone and leptin in men. AB - Besides its role in the regulation of energy balance, leptin seems to be involved in linking energy stores to the reproductive system. A gender-dependent difference exists in plasma leptin concentration and leptin messenger ribonucleic acid expression in rodents and humans. This difference does not seem to be explained simply by differences in the amount of body fat between genders. To elucidate the relationship of endogenous testosterone and leptin, we studied the serum leptin concentrations in 269 elderly nondiabetic men. In addition, to assess whether exogenously administered testosterone could influence leptin production, we followed the serum levels of leptin in 10 healthy men during a 12 month treatment with 200 mg testosterone enanthate, i.m., weekly for contraceptive purposes. We found that the serum leptin concentration correlated inversely (r = -0.39; P < 0.001) with that of testosterone in elderly men. This inverse correlation was still present when body mass index and plasma insulin were included in the analysis. The administration of testosterone to young men suppressed serum leptin from the pretreatment level of 3.4 +/- 1.4 to 1.9 +/- 0.6 micrograms/L during the therapy. After cessation of testosterone injections, serum leptin concentration returned back to the pretreatment level. It is concluded that testosterone has a suppressive effect on leptin production, as reflected by circulating levels of this hormone. PMID- 9745437 TI - Promoter characterization of the human Na+/I- symporter. AB - The Na+/I- symporter (NIS) is the plasma membrane protein that mediates active iodide uptake into thyroid follicular cells. To understand the molecular mechanisms of human NIS (hNIS) gene regulation, the 2-kb immediate 5'-flanking region of hNIS was characterized. The tentative hNIS transcriptional start site was mapped to -375 nucleotide relative to the ATG site. Various 5'-genomic DNA fragments were tested for promoter activity in thyroid and nonthyroid cells. Our results indicate that the DNA regulatory elements in the 2-kb immediate 5' flanking region were not sufficient to confer thyroid-selective transcription of the hNIS gene. In addition, hNIS minimal promoter was localized to a region of 144 bp that contains a 90-bp stretch of a highly conserved DNA fragment between hNIS and rat NIS. PMID- 9745438 TI - The insulin-like growth factor type I receptor stimulates growth and suppresses apoptosis in prostatic stromal cells. AB - Stromal cells derived from benign prostatic hyperplasia synthesize and secrete measurable levels of insulin-like growth factor (IGF). Seventy-two-hour conditioned medium obtained from these cells contains IGF-II at levels ranging from 125-177 ng/mL.10(6) cells. IGF-I is almost undetectable. RT-PCR analysis has demonstrated that the genes for both the type I IGF receptor (IGF-IR) and the type II IGF receptor (IGF-IIR) are expressed by benign stromal cells in vitro. Competition binding analysis for IGF-I and IGF-II confirmed the existence of binding sites for both ligands with respective Kd and binding capacities of 4.9 x 10(-9) mol/L and 6.6 x 10(5) sites/cell and 4.7 x 10(-9) mol/L and 3.8 x 10(6) sites/cell. Under serum-free conditions, IGF-I and IGF-II at 500 ng/mL induce 80% and 113% increases in stromal cell density, respectively, over a 96-h period. Incubation with the IGF-IR-neutralizing antibody alpha IR3 (50 micrograms/mL) reduces the rate of stromal cell proliferation by approximately 60-80% even in the presence of stimulatory concentrations of IGFs. Camptothecin-induced apoptosis is inhibited by the addition of IGF-I and -II (500 ng/mL). alpha IR3 suppresses these survival signals and itself induces cell death in the prostatic stroma. The data suggest that IGF-IR is a pivotal molecule in prostatic stromal cell maintenance, and that specific antagonism may offer a novel means of controlling the fibromuscular expansion characteristic of benign prostatic hyperplasia. PMID- 9745439 TI - Vitamin K status and bone health: an analysis of methods for determination of undercarboxylated osteocalcin. AB - Recent studies suggest that fracture risk is associated with increased undercarboxylated osteocalcin. Methods use differences in binding of undercarboxylated and fully carboxylated osteocalcin to hydroxyapatite or barium sulfate. We evaluated these methods and found that results varied with the amount and preparation of the salts. Furthermore, patient samples with differing amounts of total osteocalcin could not be directly compared. Errors in the determination of undercarboxylated osteocalcin were minimized by expressing data as the percent of the total osteocalcin in the sample, and correcting for the basal level of osteocalcin using a polynomial equation derived from multiple binding curves. Errors from 5-15% in estimation of undercarboxylated osteocalcin were observed without both of these corrections. When differing types of assays were employed (RIA, intact, N-terminal), results also were affected. In normal adults and children and in patients on long-term warfarin therapy, the percent osteocalcin not bound to hydroxyapatite was lower when measured with an intact assay than by a polyclonal RIA. Differences were related to the amount of N-terminal osteocalcin fragments, which had low affinity for hydroxyapatite and resulted in variable overestimation of undercarboxylated osteocalcin. In a kit specific for uncarboxylated osteocalcin, we found good discrimination between carboxylated and uncarboxylated intact osteocalcin. However, the assay detected large osteocalcin fragments and overestimated their concentration by up to 350%. Values for uncarboxylated osteocalcin were not different in patients on coumadin compared with normal adults with this kit, but when normalized to the total intact osteocalcin, percent uncarboxylated osteocalcin was greater in patients on coumadin than in controls, as would be expected. Kit values for uncarboxylated osteocalcin in normal children were higher than intact values in the same subject, because of the increased reactivity of the kit toward circulating fragments that were elevated in children. Thus, for estimation of undercarboxylated osteocalcin, care must be taken to standardize the hydroxyapatite or barium sulfite used for binding, to correct for the basal level of osteocalcin in the sample, to use immunoassays that do not detect small fragments, and to express the results as the percent of the total osteocalcin in the sample. Without these precautions, the assessment of undercarboxylated osteocalcin is not reliable. PMID- 9745440 TI - Metabolism of Apo(a) and ApoB100 of lipoprotein(a) in women: effect of postmenopausal estrogen replacement. AB - The metabolism in plasma of apo(a) and apoB100, the major protein components of lipoprotein(a) [Lp(a)], and the mechanism by which estrogen lowers Lp(a) concentration are both not well understood. Estrogen or placebo were administered to 12 postmenopausal women in a double-blind cross-over design; and after each treatment, apo(a) and apoB100 in Lp(a) were endogenously labeled by i.v. trideuterated leucine. After estrogen treatment, mean Lp(a) concentration decreased during estrogen, from 25 mg/dL, by 20% (P < 0.01); and the mean production rate of apo(a) decreased, from 0.31 nmol/kg.day, by 34% (P = 0.046). In contrast, the mean fractional catabolic rates of apo(a) were similar, 0.36 vs. 0.31/day (P = 0.23). In 6 women, the kinetics of apo(a) and apoB100, the two major proteins of Lp(a), were studied during estrogen and placebo periods. During both periods, the rate of appearance of tracer was similar in Lp(a)-apo(a) and Lp(a)-apoB100, as were the resulting metabolic rates and the changes during estrogen treatment. In conclusion, the findings are more compatible with intracellular synthesis of Lp(a) from nascent apo(a) and apoB100 than extracellular assembly from plasma low-density lipoproteins. Reduced flux into plasma of Lp(a), an atherogenic lipoprotein, could contribute to the lower cardiovascular disease rates in women receiving estrogen replacement therapy. PMID- 9745441 TI - Plasma adrenal, gonadal, and conjugated steroids before and after long-term overfeeding in identical twins. AB - An analysis of the data collected in the Quebec Overfeeding Study of identical twins was undertaken to determine any evidence of a genotype effect on plasma levels of adrenal and gonadal steroids arising from long term positive energy balance. Plasma levels of sex hormone-binding globulin (SHBG), testosterone, dihydrotestosterone (DHT), dehydroepiandrosterone sulfate (DHEA-S), androsterone glucuronide, androstane-3 alpha, 17 beta-diol glucuronide (3 alpha-DIOL-G), and cortisol were measured in 12 pairs of young, sedentary, male monozygotic twins before and after 100 days of overfeeding. The dietary energy excess of 4.2 MJ/day (1000 Cal), 6 days a week, resulted in a total positive energy balance of 353 MJ (84,000 Cal). Overfeeding induced significant changes (P < 0.0001) in body weight and other measures of body composition. Within-twin pair resemblance was observed at baseline in all steroids, except cortisol [intraclass correlation range: DHEA S, 0.50 (P < 0.05); DHT, 0.77 (P < 0.001)] and was lost with overfeeding, except for DHT and SHBG (P < 0.05). SHBG levels fell and 3 alpha-DIOL-G rose with the gain in body fatness. The change in testosterone was a significant correlate of the change in upper body fat (r = -0.48; P < 0.05). The change in 3 alpha-DIOL-G correlated positively with increases in all measures of central adiposity (r = 0.52; P < 0.01). A decrease in DHEA-S occurred with a higher, but not with a lower, gain in abdominal visceral fat (P < 0.05). Thus, analysis of adrenal and gonadal steroids and of conjugated metabolites before and after overfeeding in monozygous twins supports the idea that there is a genotype effect on steroid circulating steroid levels and that these blood levels are correlated with the pattern of body fat distribution. Moreover, the baseline within-twin pairs similarity in steroid levels was attenuated by prolonged positive energy balance and body fat gain. PMID- 9745442 TI - Interleukin-13 inhibits cell proliferation and stimulates interleukin-6 formation in isolated human osteoblasts. AB - Interleukin-13 (IL-13) is a recently identified cytokine that is secreted by activated T cells and regulates inflammatory responses. We have investigated the effects of IL-13 on isolated human osteoblast-like cells (hOB). IL-13 dose dependently (1-100 pmol/L) reduced the incorporation rate of [3H]thymidine in hOB cells by more than 50%. Using a cell metabolic assay as well as direct cell counting, we found that treatment with IL-13 lead to a decrease in hOB cell number. The effect was both time and dose dependent, and after 12 days of culture, treatment with IL-13 (0.1 nmol/L) caused a 70% decrease in the number of cells. Also, IL-13 increased the levels of IL-6 messenger ribonucleic acid in hOBs, as measured by ribonuclease protection assay, and stimulated secretion of IL-6 into culture supernatants. In conclusion, IL-13 inhibits cell proliferation and increases IL-6 formation in human osteoblasts. Our findings suggest that IL 13 may cause bone loss due to impaired osteoblastic growth and IL-6-induced osteoclast recruitment. PMID- 9745443 TI - Linkage analysis of candidate genes in autoimmune thyroid disease. II. Selected gender-related genes and the X-chromosome. International Consortium for the Genetics of Autoimmune Thyroid Disease. AB - Hashimoto's thyroiditis (HT) and Graves' disease (GD) are autoimmune thyroid diseases (AITD) in which multiple genetic factors are suspected to play an important role. Until now, only a few minor risk factors for these diseases have been identified. Susceptibility seems to be stronger in women, pointing toward a possible role for genes related to sex steroid action or mechanisms related to genes on the X-chromosome. We have studied a total of 45 multiplex families, each containing at least 2 members affected with either GD (55 patients) or HT (72 patients), and used linkage analysis to target as candidate susceptibility loci genes involved in estrogen activity, such as the estrogen receptor alpha and beta and the aromatase genes. We then screened the entire X-chromosome using a set of polymorphic microsatellite markers spanning the whole chromosome. We found a region of the X-chromosome (Xq21.33-22) giving positive logarithm of odds (LOD) scores and then reanalyzed this area with dense markers in a multipoint analysis. Our results excluded linkage to the estrogen receptor alpha and aromatase genes when either the patients with GD only, those with HT only, or those with any AITD were considered as affected. Linkage to the estrogen receptor beta could not be totally ruled out, partly due to incomplete mapping information for the gene itself at this time. The X-chromosome data revealed consistently positive LOD scores (maximum of 1.88 for marker DXS8020 and GD patients) when either definition of affectedness was considered. Analysis of the family data using a multipoint analysis with eight closely linked markers generated LOD scores suggestive of linkage to GD in a chromosomal area (Xq21.33-22) extending for about 6 cM and encompassing four markers. The maximum LOD score (2.5) occurred at DXS8020. In conclusion, we ruled out a major role for estrogen receptor alpha and the aromatase genes in the genetic predisposition to AITD. Estrogen receptor beta remains a candidate locus. We found a locus on Xq21.33-22 linked to GD that may help to explain the female predisposition to GD. Confirmation of these data in HT may require study of an extended number of families because of possible heterogeneity. PMID- 9745444 TI - Serum calcitonin precursors in sepsis and systemic inflammation. AB - High serum levels of the calcitonin (CT) prohormone, procalcitonin (pro-CT), and its component peptides occur in systemic inflammation and sepsis. Using two different assays, we undertook a prospective study to determine the utility of serum precalcitonin peptides (pre-CT) as markers in this condition. Twenty-nine patients meeting criteria for the systemic inflammatory response syndrome were studied daily in two intensive care units. Sera were collected, and APACHE II scores were determined until recovery or death. All patients had markedly elevated serum pre-CT. Prognostically, peak values were the most important. The highest values portended mortality, and a lower level could be ascertained below which all patients survived. Peak pre-CT levels were significantly higher in patients with infection documented by blood cultures than in those patients with no documented infection from any source (P < 0.05). Mature CT remained normal or only moderately elevated. Compared with the serum pre-CT levels, receiver operating characteristic curve analysis revealed that the APACHE II scores, although more cumbersome, were better overall predictors of mortality. Thus, pre CT is an important serum marker for systemic inflammatory response syndrome and is predictive of outcome. It also provides data concerning the presence of severe infection and may prove to be clinically useful for proactive patient care. PMID- 9745445 TI - Net increase in stimulatory input resulting from a decrease in inhibin B and an increase in activin A may contribute in part to the rise in follicular phase follicle-stimulating hormone of aging cycling women. AB - Recent studies suggest that an age-related decline in ovarian inhibin B may play a role in the increase in follicular phase FSH in menstrual cycles of older women. Considering that the peripheral feedback regulation of FSH is dictated by the overall tone of inhibins, activins, and follistatins as well as estradiol, it is essential to determine the relative inputs of all of these regulators in assessing whether the collective peripheral input to FSH is one of inhibition or stimulation. To test the hypothesis that changes in the overall tone of peripheral feedback may contribute to this hallmark sign of aging, we compared the concentrations of dimeric inhibin A, inhibin B, activin A, and total and free follistatin in 7 young (mean age, 27.9 +/- 2.6 yr) and 10 older (mean age, 43.6 +/- 0.9 yr) cycling women during the follicular (FOLL; cycle day 6) and midluteal (ML; 7 days post-LH surge) phases of the menstrual cycle. Subjects were preselected on the basis of FOLL phase FSH levels (older, > or = 8.0 mIU/mL; younger, < 8 mIU/mL). Circulating FSH regulatory peptide concentrations were determined from samples pooled from blood drawn every 10 min for 8 daytime h using specific 2-site assays. In the older group, cycle length was shorter (29.1 +/- 0.5 vs. 26.1 +/- 0.5, young vs. older; P < 0.001), mean LH levels during the follicular phase were higher (LH, 5.6 +/- 0.8 vs. 8.8 +/- 1.1 mIU/mL, young vs. older; P < 0.001). Mean FSH levels for the older and younger groups averaged 10.8 +/- 0.8 and 6.2 +/- 0.3 mIU/mL, respectively. Estradiol levels were higher, but not statistically different, than those in the younger group (99 +/- 13 vs. 169 +/- 25 pmol/L, young vs. older; P = 0.06). In both age groups, inhibin B levels were higher in the FOLL vs. ML phase, inhibin A levels were higher in the ML vs. FOLL phase, but total activin A and total and free follistatin did not differ across cycle days. FOLL phase inhibin A levels were higher in the older group (16.3 +/- 2.4 vs. 26.4 +/- 3.4 pg/mL, young vs. older; P = 0.024), but levels of inhibin B were lower (323 +/- 80 vs. 163 +/- 24 pg/mL, young vs. older; P = 0.03). Overall, the estimated total inhibin activity (inhibin A plus inhibin B) was lower in older cycling than in younger women (339 +/- 82 and 189 +/- 24 pg/mL, young vs. older). Total and free follistatin levels were not different among the 2 groups of women. In contrast, total activin A levels were higher in the older cycling group (0.51 +/- 0.05 and 0.68 +/- 0.05 ng/mL, young vs. older; P = 0.02). No differences in age groups were observed during the ML phase for any of the variables measured. These data suggest that a net increase in stimulatory input resulting from a decrease in inhibin B and an increase in activin A may contribute in part to the monotropic FSH increase in aging women. PMID- 9745446 TI - Differential expression of estrogen receptor-beta (ER beta) in human pituitary tumors: functional interactions with ER alpha and a tumor-specific splice variant. AB - The mitogenic and regulatory effects of estrogen (E2) in adenohypophysial cells are known to be mediated through the nuclear estrogen receptor (ER alpha). Expression of ER alpha and several of its messenger ribonucleic acid (RNA) alternate splice variants has been shown to be restricted to prolactinomas and gonadotroph tumors. However, little is known about gene expression patterns of the novel nuclear hormone receptor ER beta in the neoplastic pituitary. ER beta has high homology to ER alpha in the DNA- and ligand-binding domains, but encodes a distinct transcriptional activating function-1 (AF-1) domain. Using RT-PCR analysis of total RNA from 38 human pituitary adenomas, we found that ER beta messenger RNA was coexpressed with ER alpha and its splice variants in 60% of prolactinomas, 100% of mixed GH/PRL tumors, and 29% of gonadotroph tumors. ER beta gene expression was not limited to ER alpha-positive tumor subtypes, however, and was also found in 100% of null cell tumors, 80% of somatotroph tumors, and 60% of corticotroph tumors. Because ER beta is coexpressed with ER alpha and its splice variants in prolactinomas and gonadotroph tumors, we functionally characterized the potential interactions between ER beta and ER alpha. We also examined the potential cooperative effects on ER beta-mediated gene expression of a tumor-specific truncated delta 5ER alpha splice variant that has been shown to be coexpressed in the majority of ER alpha-positive tumors. This exon 5 splice variant encodes the AF-1 domain as well as regions critical for DNA binding and nuclear localization, but lacks the ligand-binding and AF-2 domains. Mammalian expression vectors encoding ER alpha, delta 5ER alpha, and/or ER beta complementary DNAs were transiently transfected along with an E2 response element promoter-luciferase (ERELuc) reporter into human ER alpha/ER beta negative osteosarcoma U2-OS cells. ER beta was less potent than ER alpha in activating E2-stimulated ERELuc activity (4-vs. 14-fold relative to basal control levels). However, when delta 5ER alpha was coexpressed with ER beta or ER alpha, E2-stimulated ERELuc activity was markedly increased to 8- and 57-fold, respectively, relative to basal control levels when each full-length isoform was expressed alone. Finally, coexpression of ER beta with ER alpha did not significantly alter the E2-stimulated ERELuc activity induced by ER alpha alone. Cotreatment with tamoxifen markedly inhibited all E2-stimulated ERELuc responses to baseline levels. Together, these data suggest that ER beta has a minor role in mediating E2 responses in ER alpha-positive tumors, but may be the main mediator of E2-stimulated gene expression when expressed alone in somatotroph, corticotroph, and null cell tumors. This low, but significant, level of ER beta trans-activation potential may be enhanced by coexpression of delta 5ER alpha in neoplastic pituitary. Therefore, E2-mediated gene expression in normal and neoplastic pituitary appears to be highly dependent on the expression of ER alpha and ER beta isoforms, which have varying transcriptional activities. PMID- 9745447 TI - Insulin-like growth factor binding protein-3: a novel biomarker for the assessment of the synthetic capacity of hepatocytes in liver cirrhosis. AB - The liver is the major source of circulating insulin-like growth factor binding protein-3 (IGFBP-3). Because the hepatic tissue is deranged in cirrhotic patients, we measured serum IGFBP-3 concentrations by two-site immunoradiometric assay in sera from 37 cirrhotic patients with different stages of hepatic dysfunction. These were compared with IGFBP-3 levels from 11 healthy controls. Serum IGFBP-3 levels in patients with chronic liver disease were significantly lower than those of the control group (P < 0.0005). The mean percent decrease in cases of early liver cirrhosis, cirrhosis without, and cirrhosis with ascites were 44%, 59%, and 82% respectively, indicating that serum IGFBP-3 levels decrease as the severity of hepatic dysfunction increases. Moreover, the decrease was more pronounced in cases with hyperbilirubinemia, elevated serum transaminases, hypoalbuminemia, and prolonged prothrombin time. There was a significant positive correlation between serum IGFBP-3 and serum albumin, as well as a significant negative correlation between serum IGFBP-3 and prothrombin time. These results indicate the close correlation of IGFBP-3 levels to worsening of hepatic functions. The determination of serum IGFBP-3 level is a clinically useful marker for the assessment of the synthetic capacity of hepatocytes in cirrhotic patients and an early predictor of impending hepatic dysfunction as well. PMID- 9745448 TI - Neutral amino acid uptake by the microvillous plasma membrane of the human placenta is inversely related to fetal size at birth in normal pregnancy. AB - Understanding the physiological regulation of fetal growth is important, as normal variations in size at birth relate to differences in neonatal and adult health. Although fetal growth directly reflects net placental transfer, little is known about how normal fetal growth relates to the transfer capabilities of the placental epithelium, the syncytiotrophoblast. The Na(+)-dependent and Na(+) independent uptakes of methylaminoisobutyric acid (MeAIB) by vesicles prepared from the syncytiotrophoblast microvillous plasma membrane give measurements of system A neutral amino acid transporter activity and diffusive permeability, respectively. In 62 normal pregnancies, we related vesicle MeAIB uptakes to neonatal anthropometry. Smaller babies with a lower abdominal circumference had higher placental system A activity per mg membrane protein (P = 0.004); activity rose from 0.020 to 0.043 nmol/30 sec/mg protein as abdominal circumference fell from 34.6 cm or more to 32.0 cm or less. Within the normal range of fetal and placental size, this may reflect a tendency toward compensatory up-regulation of the placental system A transporter in smaller babies. Babies with a lower abdominal circumference also had higher Na(+)-independent MeAIB uptakes (P = 0.0005); this could reflect important compositional changes in the microvillous plasma membrane, leading in vivo to increased back-diffusion of amino acids out of the syncytiotrophoblast. PMID- 9745449 TI - Mutation and expression analysis of corticotropin-releasing factor 1 receptor in adrenocorticotropin-secreting pituitary adenomas. AB - The present study was designed to investigate a possible role of CRF1 receptors (CRF1-R) in the pathogenesis of Cushing's disease. ACTH-secreting pituitary adenomas and nonsecreting pituitary adenomas have been analyzed for mutations in the CRF1-R gene by PCR and sequencing and been compared with the sequences of normal anterior pituitaries. No mutations affecting the CRF1-R protein have been found in all tumors analyzed. However, we found a significant overexpression of the CRF1-R messenger RNA in ACTH-secreting pituitary adenomas vs. inactive adenomas and normal pituitaries. We conclude that mutations of the CRF1-R are unlikely to be involved in Cushing's disease. We suggest that the overexpression of the CRF1-R messenger RNA may be related to a disturbed receptor regulation in ACTH-secreting pituitary adenomas. PMID- 9745450 TI - High intensity exercise promotes escape of adrenocorticotropin and cortisol from suppression by dexamethasone: sexually dimorphic responses. AB - Exercise promotes escape of ACTH and cortisol from suppression by dexamethasone (DEX) in some healthy men and women. To determine whether stimulus strength, diurnal rhythmicity, or gender influences neuroendocrine escape during DEX suppression, we studied men (n = 5) and women (n = 5) during high intensity exercise tests after taking 4 mg DEX: two tests (one at 90% and one at 100% of maximal aerobic capacity) were conducted in the morning and two were performed in the afternoon on nonconsecutive days. Plasma ACTH and cortisol showed significantly greater increases with the 100% compared to the 90% intensity exercise (ACTH: 90%, 2 +/- 0.4; 100%, 3 +/- 0.5 pmol/L; cortisol: 90%, 53 +/- 5.3; 100% 93 +/- 23.6 nmol/L). Plasma cortisol responses were significantly higher in women than in men (P < 0.01). Plasma arginine vasopressin (AVP) exhibited significant intensity-dependent increases, with higher responses in women than men (P < 0.01). In conclusion, despite high dose glucocorticoid pretreatment, intense exercise can override the glucocorticoid negative feedback of hypothalamic-pituitary-adrenal activation in most normal men and women. This ability to override cortisol negative feedback inhibition may relate to the magnitude of the AVP response, the potency/specificity of the stressor to elicit a CRH/AVP response, and/or the sensitivity of the glucocorticoid negative feedback system at the time of the stress. PMID- 9745451 TI - The role of mineralocorticoid receptors in hypothalamic-pituitary-adrenal axis regulation in humans. AB - In rodents, two types of glucocorticoid receptors, the mineralocorticoid (MR; type I) and the glucocorticoid (type II) receptors, have been demonstrated to play a role in hypothalamic-pituitary-adrenal (HPA) axis regulation. Because MR shows a very high affinity for cortisol, it has been suggested that MR plays an important role in restraint of CRH and ACTH secretion during the nadir of the circadian rhythm. Although a number of studies have established the importance of MR in rodents, the functional role of MR in humans has not been determined. These studies evaluated whether spironolactone, an MR antagonist, had a detectable effect on HPA axis regulation in humans, and whether the effect was greatest during the evening, when plasma cortisol concentrations are in the MR range. Compared to the placebo day, after a single dose of spironolactone at either 0800 or 1600 h, there is a significant increase in plasma cortisol, which is preceded by a rise in ACTH and beta-endorphin. A significant effect of spironolactone on cortisol secretion was demonstrated with no differences between the morning and evening. Because the effect of spironolactone on cortisol was short lived, a second experiment was conducted using two doses of spironolactone, again sampling in the morning and evening. After two doses of spironolactone, plasma cortisol levels showed a significant and sustained spironolactone-induced elevation for the entire sampling period. However, neither plasma beta-endorphin nor ACTH was increased compared to levels on the placebo day. These data suggest that MR appear to play a clear role in HPA axis regulation during the time of the circadian peak as well as the trough. Furthermore, MR blockade may affect the sensitivity of the adrenal to ACTH. PMID- 9745453 TI - Age-related reduction in glucose elimination is accompanied by reduced glucose effectiveness and increased hepatic insulin extraction in man. AB - This study examined whether insulin secretion, insulin sensitivity, glucose effectiveness (SG), and hepatic extraction (HE) of insulin are altered by age when glucose tolerance is normal. A frequently sampled i.v. glucose tolerance test was performed in 20 elderly (E, 10/10 male/female, all 63 yr old) and in 20 young subjects (Y, 10/10 male/female, all 27 yr old), who were similar in body mass index and 2-h blood glucose during oral glucose tolerance test. E exhibited impaired glucose elimination (i.v. tolerance index, 1.31 +/- 0.10 vs. 1.70 +/- 0.12% min-1; P = 0.019). First-phase insulin secretion and SI did not differ between the groups, whereas E had lower glucose sensitivity of second-phase insulin secretion (0.40 +/- 0.07 vs. 0.70 +/- 0.08 (pmol/L)min-2/(mmol/L), P = 0.026), lower SG, 0.017 +/- 0.002 vs. 0.025 +/- 0.002 min-1, P = 0.004), and higher HE (81.3 +/- 2.4 vs. 73.2 +/- 2.1%, P = 0.013). Across both groups, SG correlated positively with glucose tolerance index (r = 0.58, P < 0.001) and negatively with HE (r = -0.54, P < 0.001). Plasma leptin and glucagon did not change by age, whereas plasma pancreatic polypeptide (PP) was higher in E (122 +/ 18 vs. 66 +/- 6 pg/mL, P = 0.004). PP did not, however, correlate to any other parameter. We conclude that E subjects with normal oral glucose tolerance have reduced SG, impaired second-phase insulin secretion, and increased HE, whereas SI and first-phase insulin secretion seem normal. SG seems most related to age dependent impairment of glucose elimination, whereas leptin, glucagon, and PP do not seem to contribute. PMID- 9745452 TI - The PROP1 2-base pair deletion is a common cause of combined pituitary hormone deficiency. AB - Combined pituitary hormone deficiency (CPHD) has an incidence of approximately 1 in 8000 births. Although the proportion of familial CPHD cases is unknown, about 10% have an affected first degree relative. We have recently reported three mutations in the PROP1 gene that cause CPHD in human subjects. We report here the frequency of one of these mutations, a 301-302delAG deletion in exon 2 of PROP1, in 10 independently ascertained CPHD kindreds and 21 sporadic cases of CPHD from 8 different countries. Our results show that 55% (11 of 20) of PROP1 alleles have the 301-302delAG deletion in familial CPHD cases. Interestingly, although only 12% (5 of 42) of the PROP1 alleles of our 21 sporadic cases were 301-302delAG, the frequency of this allele (in 20 of 21 of the sporadic subjects given TRH stimulation tests) was 50% (3 of 6) and 0% (0 of 34) in the CPHD cases with pituitary and hypothalamic defects, respectively. Using whole genome radiation hybrid analysis, we localized the PROP1 gene to the distal end of chromosome 5q and identified a tightly linked polymorphic marker, D5S408, which can be used in segregation studies. Analysis of this marker in affected subjects with the 301 302delAG deletion suggests that rather than being inherited from a common founder, the 301-302delAG may be a recurring mutation. PMID- 9745454 TI - Salt-wasting congenital adrenal hyperplasia: detection of mutations in CYP21B gene in a Chilean population. AB - The steroid 21-hydroxylase deficiency (21OHD) is the most frequent cause of congenital adrenal hyperplasia. We have characterized the disease-causing mutations in the 21-hydroxylase genes of 63 patients with salt-wasting congenital adrenal hyperplasia from a Chilean population of Hispanic origin, a group that has been scarcely evaluated. Using allele-specific PCR, lesions were identified in 97 chromosomes out of 126 tested (77%). The most frequent findings were the gene deletion or large gene conversion (LGC) = 22.9%, I2 splice = 19%, R357W = 12.7%, and Q319X = 10.5%. We did not find alleles with the mutation F308insT and we found three alleles with the cluster E6. The frequency of the point mutation R357W was at least two times more frequent than the one found in Caucasians populations, but similar to that communicated in Asian populations; this finding may be explained by the Asian ancestry of our South-Amerindian population. The frequency of Q319X was also high, similar only to those patients studied in Italy and in a neighboring Argentinian population. In summary, this is a genetic characterization of 21OHD made in an almost pure Hispanic population in Latin America. The high frequency of deletion of CYP21B gene, I2 splice, R357W, and Q319X mutations probably reflects the European-Caucasian-Spanish influence of the conquerors, mixed with Amerindians of Asian ancestry and modulated by other European immigrations. PMID- 9745455 TI - Molecular analysis of the ret and GDNF genes in a family with multiple endocrine neoplasia type 2A and Hirschsprung disease. AB - The clinical association between multiple endocrine neoplasia type 2 (MEN2) and Hirschsprung disease (HSCR) is infrequent. Germline mutations of the ret protooncogene are the underlying cause of the MEN2 syndromes and a proportion of cases of HSCR. In this report, we describe a new kindred in which the MEN2 and HSCR phenotypes are associated with a single C620S point mutation at one of the cysteine codons of the extracellular domain of the ret protooncogene. We also speculate about the role of a silent mutation in exon 2 of this same gene (A45A), present in a homozygous state in the patient with both MEN2A and HSCR. To investigate the contribution of GDNF to the phenotype observed in this kindred, we scanned the coding region of GDNF in the patient with MEN2/HSCR, but no mutation was found. PMID- 9745456 TI - A homozygous inactivating mutation in the parathyroid hormone/parathyroid hormone related peptide receptor causing Blomstrand chondrodysplasia. AB - We describe a patient with Blomstrand chondrodysplasia, a lethal genetic disorder characterized by extremely advanced endochondral bone maturation, in whom a homozygous missense mutation is present in the gene coding for the PTH/PTHrP receptor that leads to the substitution of a proline for a leucine in the N terminal portion of the receptor (P132L). PTH-induced cAMP accumulation was severely reduced in COS-7 cells expressing P132L receptors compared to that of cells expressing wild-type receptors, and PTH-induced inositol phosphate accumulation was not detectable in cells expressing the mutant receptor. Similar results were obtained using PTHrP as an agonist. Maximal specific binding of radioiodinated [Tyr36]PTHrp(1-36) by cells transfected with the P132L receptor was < 10% of that observed for cells transfected with the wild-type receptor. Despite the reduction in radioligand binding to P132L receptors, the intensity and distribution of the fluorescent signal resulting from the expression of receptors fused to GFP were similar for cells transfected with the wild-type and mutant P132L receptors, suggesting a similar degree of cell surface expression. These results firmly establish the role of abnormalities in the PTH/PTHrP receptor in the pathogenesis of Blomstrand chondrodysplasia, and thereby confirm the importance of signaling through the PTH/PTHrP receptor in human fetal skeletal development. Because the amino-acid mutated in the patient described here is otherwise conserved in all mammalian class II G protein-coupled receptors, this abnormality may provide insights into structural features needed for the normal function of this family of receptors. PMID- 9745457 TI - Extraocular light exposure does not suppress plasma melatonin in humans. AB - Light affects the circadian axis in at least two ways. It can cause the acute suppression of pineal melatonin synthesis, and/or a phase-shift of the circadian oscillator. As recent evidence has suggested that extraocular light exposure may cause phase-shifts of the circadian clock, we have investigated whether suppression of melatonin can be induced by the same type of light exposure. In the first study subjects' eyes were exposed to white light (2250 lux for 30 mins) via a fibre optic cable. As expected, suppression of nighttime plasma melatonin levels (61 +/- 6%) was observed. In the second study, light of the same quality but higher intensity (14,000 or 67,500 lux for 180 mins) was delivered in the same manner to the popliteal region behind the subjects' knees, whilst shielding their eyes. No suppression of plasma melatonin levels (4 +/- 7%) was detected in any of the subjects. Thus, extraocular photoreception, if it exists in mammals, does not affect the suprachiasmatic nucleipineal pathway. PMID- 9745458 TI - Novel, missense and loss-of-function mutations in the sodium/iodide symporter gene causing iodide transport defect in three Japanese patients. AB - Iodide transport defect is a disorder affecting the active transport of iodide, an essential step in the synthesis of thyroid hormones. We have identified novel germ-line mutations in the Na+/I- symporter (NIS) gene from three Japanese patients with iodide transport defect. One patient had a compound heterozygous mutation of T354P/G93R (Gly93-->Arg [GGC-->CGC]), and two sibling patients had a homozygous mutation of G543E (Gly543-->Glu [GGA-->GGA]). G93R and G543E, two novel mutations, are located in the 3rd and 12th transmembrane domains of NIS which are encoded by exons 1 and 13, respectively. The NIS mutants carrying these mutations had minimal iodide uptake activity when expressed in COS-7 cells, confirming that the identified mutations are the direct cause of the iodide transport defect in these patients. Genotyping of unaffected family members and functional assays of co-transfected COS-7 cells indicate that expression of one normal NIS allele in the heterozygote (T354P, G93R, or G543E) is sufficient to maintain active iodide uptake activity. Thus, none of these NIS mutants acts as a dominant-negative mutant. PMID- 9745459 TI - Effect of thyroxine on low density lipoprotein oxidation another thyroid hormone nongenomic effect. PMID- 9745460 TI - Comment on high urinary free cortisol excretion in a patient with psychogenic polydipsia. PMID- 9745461 TI - Prolonged suppression of corticosteroid-binding globulin by recombinant human interleukin-6 in man. PMID- 9745462 TI - Comment on Therapeutic controversy: Thyroid surgery--the choice. PMID- 9745463 TI - In memoriam: Roy Orval Greep. PMID- 9745464 TI - Quality of life after intestinal transplantation and on total parenteral nutrition. PMID- 9745465 TI - Long-term nutritional monitoring after clinical intestinal transplantation. PMID- 9745466 TI - Nutritional management of intestinal allograft recipients. PMID- 9745467 TI - Nutritional monitoring of pediatric intestinal transplant recipients. PMID- 9745468 TI - Results of pediatric small bowel transplantation in Canada. PMID- 9745469 TI - Intestinal failure in children. PMID- 9745470 TI - Preliminary experience with combined liver and small bowel transplantation in children. PMID- 9745471 TI - Post-absorptive plasma citrulline concentration: a marker of intestinal failure in humans. PMID- 9745472 TI - Potential candidates for small bowel transplantation: from our experience and survey of home parenteral nutrition in Japan. PMID- 9745473 TI - Demand for pediatric small bowel transplantation in the United Kingdom. PMID- 9745474 TI - Experience with combined liver-small intestine transplantation at the University of California, Los Angeles. PMID- 9745475 TI - Intestinal motility after small bowel transplantation. PMID- 9745476 TI - Non-tuberculous mycobacterial associated enterocolitis in intestinal transplantation. PMID- 9745477 TI - Long-term parenteral nutrition in the management of extremely short bowel syndrome. PMID- 9745478 TI - Intractable diarrhea caused by intestinal fibrosis. PMID- 9745479 TI - Long-term parenteral nutrition and parenteral nutrition dependency in pediatric patients. PMID- 9745480 TI - First-pass metabolism of cyclosporine A in human intestine: inhibition by diltiazem. PMID- 9745481 TI - Severe liver complications associated with long-term parenteral nutrition are dependent on lipid parenteral input. PMID- 9745482 TI - Long-term survival and parenteral nutrition-dependency of adult patients with nonmalignant short bowel. PMID- 9745483 TI - Elevated immunoreactive trypsinogen levels in pediatric patients with short bowel syndrome. PMID- 9745484 TI - The role of the small bowel as a barrier for bacterial septic shock. PMID- 9745486 TI - Immunologic aspects of small bowel transplantation. PMID- 9745485 TI - Microchimerism is associated with long-term graft acceptance in combined liver/small bowel transplantation. PMID- 9745487 TI - Immunology of the small intestine. PMID- 9745488 TI - Increased bacterial clearance with portal bacterial challenge compared with systemic bacterial challenge. PMID- 9745489 TI - Clearance of lethal and nonlethal concentrations of bacteria from systemic circulation. PMID- 9745490 TI - Tissue-specific immune response: the role of innate immunity in small bowel and heart transplantation. PMID- 9745491 TI - Role of phagocytes in causing dysmotility after each stage of small bowel transplantation. PMID- 9745492 TI - Extracellular matrix: an early target of preservation/reperfusion injury and acute rejection after small bowel transplantation. PMID- 9745493 TI - Tolerance induction with peripheral blood lymphocyte depletion using monoclonal antibodies: tip of the iceberg? PMID- 9745495 TI - A second intrathymic injection of donor splenocytes improves survival of small bowel allografts. PMID- 9745494 TI - Tolerance: a study in long-term surviving animals after small bowel transplantation. PMID- 9745496 TI - Effect of perioperative donor bone marrow infusion after small bowel transplantation in swine: preliminary results. PMID- 9745497 TI - Combined effect of rapamycin and FK 506 in prolongation of small bowel graft survival in the mouse. PMID- 9745498 TI - Mesenteric alterations in pigs with chronically rejecting small bowel allografts. PMID- 9745499 TI - Induction of heat-shock protein protects the small intestine from preservation injury. PMID- 9745500 TI - Histopathologic changes during chronic small bowel allograft rejection. PMID- 9745502 TI - Apoptosis in combined liver/small bowel transplantation. PMID- 9745501 TI - Upregulation of basic fibroblast growth factor during chronic rejection of rat intestinal allografts and its downregulation by FK 506. PMID- 9745503 TI - Rat-to-mouse small bowel xenotransplantation: novel models to study hyperacute and acute humoral rejection. PMID- 9745504 TI - Graft survival and MLC response in LEW-ACI small bowel transplants treated with a novel MHC peptide, BC-1nl. PMID- 9745505 TI - Role of CD8+ and CD4+ T cells in the rejection of murine intestinal allografts. PMID- 9745506 TI - GvHD after small bowel transplantation: the role of caspases, FAS-L, and galectin 1. PMID- 9745507 TI - Prolongation of rat small bowel allograft survival by CTLA-4 IG. PMID- 9745508 TI - Increase of interleukin-6 in the intestinal mucosa and muscularis following chronic rejection after small bowel transplantation. PMID- 9745509 TI - Graft-versus-host reaction and graft rejection after liver, small bowel or small bowel allotransplantation in the pig. PMID- 9745510 TI - Effect of selective immunosuppression with FK 506, anti-IL-2R, and anti-ICAM-1 MAb in rat small bowel transplantation. PMID- 9745511 TI - Clinical observation of intestinal transplantation with microsurgical lymphatic vessel and nerve reconstruction in the allogeneic orthotopic rat model. PMID- 9745512 TI - Development of a model of combined bone marrow and small bowel transplantation in swine. PMID- 9745514 TI - Development of a porcine small bowel ex vivo perfusion model. PMID- 9745513 TI - A model of myoelectrical activity recording after small bowel transplantation. PMID- 9745515 TI - Changes in plasma endotoxin levels after small bowel auto/allo-transplantation in pigs. PMID- 9745516 TI - Splanchnic jump graft technique with passive venovenous bypass improves survival of multivisceral transplantation in a pig model. PMID- 9745517 TI - Technical refinement using bulldog clamp for vascular anastomosis in rat small bowel transplantation. PMID- 9745518 TI - Effect of epidermal growth factor on small bowel transplantation in rats. PMID- 9745519 TI - Distribution in nitric oxide neurons after rat small intestinal transplantation. PMID- 9745520 TI - Multivisceral transplantation in pigs: technical aspects. PMID- 9745521 TI - Large animal models in intestinal transplantation. PMID- 9745522 TI - Synchronous intestinal transplantation inhibits post resection adaptation. PMID- 9745523 TI - Comparison of jejunal and ileal grafts from the aspects of cold preservation and immunologic reaction. PMID- 9745525 TI - Structural assessment of intestinal grafts preserved with phospholipase A2 inhibition. PMID- 9745524 TI - Protective effect of TNF-alpha and IL-1 beta inhibitor FR167653 on ischemia reperfusion injury in rat small intestinal transplantation. PMID- 9745526 TI - Is the graft too big or too small? Technical variations to overcome size incongruity in visceral organ transplantation. PMID- 9745527 TI - Beneficial effects of microsurgical lymphatic reconstruction after intestinal transplantation in rats. PMID- 9745528 TI - Early secretory events during intestinal graft preservation. PMID- 9745529 TI - Anti-rat IL-8 (CINC) monoclonal antibody administration reduces ischemia reperfusion injury in small intestine. PMID- 9745530 TI - Vascular thrombosis in small bowel transplantation: a comparative study in the pig. PMID- 9745531 TI - Activation of STAT proteins following ischemia reperfusion injury demonstrates a distinct IL-6 and G-CSF mediated profile. PMID- 9745532 TI - Enteric nervous system in preservation, reperfusion, and rejection of the pig small bowel. PMID- 9745533 TI - Liposomal tacrolimus and intestinal drug concentration. PMID- 9745534 TI - Efficacy of cytologic examination of luminal exudate in monitoring acute rejection of rat intestinal allografts. PMID- 9745535 TI - Neoral increases bioavailability versus oral Sandimmune after porcine small bowel transplantation. PMID- 9745536 TI - Differentiation between preservation reperfusion injury and acute rejection after small bowel transplantation. PMID- 9745537 TI - Noninvasive videomicroscopic monitoring of rat small bowel rejection. PMID- 9745538 TI - Nitric oxide production after syngeneic and allogeneic small bowel transplantation. PMID- 9745539 TI - Myoelectrical activity pattern in rat Thiry-Vella loops. PMID- 9745540 TI - Nifedipine prevents tacrolimus-induced intestinal hemodynamic and functional impairments. PMID- 9745541 TI - Lipopolysaccharide activates jejunal muscularis macrophages and suppresses circular muscularis activity. PMID- 9745542 TI - Somatostatine does not decrease intestinal necrosis in an ischemia-reperfusion experimental model. PMID- 9745543 TI - Allopurinol and N-acetylcysteine avoid 60% of intestinal necrosis in an ischemia reperfusion experimental model. PMID- 9745544 TI - Apoptosis in gut-associated lymphoid tissue: a response to injury or a physiologic mechanism? PMID- 9745545 TI - Mycophenolate mofetil as primary and rescue therapy in intestinal transplantation. PMID- 9745546 TI - Intestinal transplantation in children after primary liver transplantation. PMID- 9745547 TI - Diabetogenic activity of xanturenic acid determined by its chelating properties? PMID- 9745549 TI - An open proposal for clinical composite tissue allotransplantation. PMID- 9745548 TI - Spleen transplantation from mother to child induces prolonged immunotolerance to intestinal transplantation in rats. PMID- 9745550 TI - Update and outline of the experimental problems facing clinical composite tissue transplantation. PMID- 9745551 TI - Mixed chimerism for the induction of tolerance: potential applicability in clinical composite tissue grafting. PMID- 9745552 TI - Immunomonitoring after human limb allotransplantation. PMID- 9745553 TI - Hematopoietic chimerism, tolerance induction and graft-versus-host disease: considerations for composite tissue transfer. PMID- 9745555 TI - Is histocompatibility testing needed for composite tissue transplantation? PMID- 9745554 TI - Withdrawal of immunosuppression: implications for composite tissue allograft transplantation. AB - Complete or partial withdrawal of immunosuppression is a desirable goal for physicians managing solid organ transplant recipients and has particular appeal for the management of composite tissue allograft recipients. Experience to date with steroid withdrawal or cyclosporine withdrawal in organ transplant recipients suggests that the risks of acute rejection are minimized with slow tapering of the drugs and when drug withdrawal is attempted many months or years after transplantation. Unfortunately, the full benefits of withdrawing any component of a multidrug immunosuppression regimen can probably be achieved only when the drug is withdrawn relatively early after transplantation. Thus, there is a need for improved immunologic monitoring to facilitate withdrawal of immunosuppression in any setting. Because steroid withdrawal might be particularly advantageous to the recipient of a composite tissue allograft, further experience is needed to determine the safety of steroid withdrawal with newer immunosuppressants such as tacrolimus, mycophenolate mofetil, and sirolimus. PMID- 9745556 TI - Tissue modifications. PMID- 9745557 TI - Donor and recipient immunomodulation as an aid for limb transplantation. PMID- 9745558 TI - The case for local immunosuppression in composite tissue allotransplantation. PMID- 9745559 TI - Use of swine model in transplantation of vascularized skeletal tissue allografts. AB - Permanent tolerance to vascularized skeletal tissue allografts can be induced in miniature swine with minor antigen differences using a 12-day course of CsA. Demonstration of skeletal tissue allograft survival in a large animal model without long-term immunosuppression represents an important step toward transplantation of skeletal tissue allografts in humans. PMID- 9745560 TI - Transplantation of vascularized allogeneic skeletal muscle for scalp reconstruction in renal transplant patient. PMID- 9745561 TI - Allogeneic vascularized transplantation of human femoral diaphyses and total knee joints--first clinical experiences. AB - This article has presented the preliminary results of three patients who received vascularized allogeneic femoral diaphyses and three patients having undergone vascularized transplantation of fresh and perfused total human knee joints. The large osseous defects in the femora followed osteomyelitis and chondrosarcoma. The three knee joints were lost due to various trauma mechanisms. All grafts were harvested within 25 hours from multiorgan donors perfused with 4 L of UW solution. All osteosyntheses were performed employing intramedullary nails. Vascular pedicles of the grafts were anastomosed end-to-side to the superficial femoral artery and vein in the adductorial canal of the recipient thigh. Immunosuppression was based mainly on two drugs: CyA and AZA. Perfusion of the grafts was demonstrated by DSA, and bone metabolism in the graft by SPECT scintigraphy. Six months after the operation all osteotomies demonstrated callus formation and osseous consolidation in conventional radiographs. Biopsies of the grafted bone revealed intact osteocytes, and arthroscopy of the transplanted knee joints demonstrated intact synovial, chondral, and ligamentous structures. From the surgical aspect, the vascularized transplantation of the femoral diaphyses and total knee joints is technically feasible. The main problems are immunologic. All transplantations were performed with respect to ABO compatibility, but with a large HLA mismatch. Therefore, acute and chronic rejection crises were observed. In total synovial joints, lifelong immunosuppression of graft recipients seems to be currently unavoidable. PMID- 9745562 TI - Peripheral nerve allografting: review of the literature with relevance to composite tissue transplantation. PMID- 9745563 TI - Malignancies complicating organ transplantation. PMID- 9745564 TI - Posttransplantation skin cancer: scope of the problem, management, and role for systemic retinoid chemoprevention. AB - Long-term immunosuppression necessary for transplantation places susceptible individuals with chronic actinic damage at increased risk for the development of aggressive skin cancers. Candidates for transplantation should be evaluated for the risk factors associated with skin cancer. Those who are at risk should be educated in the measures for and the importance of diligent UVR protection, frequent self-examination of the skin, and regular dermatologic evaluation to minimize the morbidity and morality from NMSC. Identified skin cancers (especially SCC) should be treated aggressively. Individuals who are actively developing many cancers may be candidates for retinoid chemoprevention. PMID- 9745565 TI - Ethical issues in innovative surgery: should we attempt a cadaveric hand transplantation in a human subject? PMID- 9745566 TI - Using decision analysis to aid in the introduction of upper extremity transplantation. PMID- 9745567 TI - Complications of pancreas transplantation: radiological evaluation. PMID- 9745568 TI - Influence of different colloids on hemodynamic and renal functions: comparative study in an isolated perfused pig kidney model. PMID- 9745569 TI - Beneficial effects of low-potassium and polyethylene glycol solution on renal lipid peroxidation during 48-hour cold storage and normothermic reperfusion. PMID- 9745570 TI - T-cell flow-cytometry cross-match: influence in renal transplantation. PMID- 9745571 TI - Economic relevance of cyclosporine microemulsion in kidney transplanted patients. PMID- 9745572 TI - Risk factors of chronic rejection after renal transplantation: retrospective study of 201 patients. PMID- 9745573 TI - Use of logistic function to analyse delayed graft function in renal transplantation. PMID- 9745574 TI - Future of children of transplanted mothers: results of a multicenter study. PMID- 9745575 TI - Contribution of mycophenolate mofetil after withdrawal of cyclosporine to toxicity in kidney transplantation. PMID- 9745576 TI - Rejection therapy with tacrolimus in renal transplantation: preliminary results of a collaborative multicenter study in 45 patients. Groupe Cooperatif de Transplantation D'ile de France (GCIF). PMID- 9745577 TI - Vascular microthrombosis in renal transplant recipients treated with tacrolimus. PMID- 9745578 TI - Loss of graft by chronic rejection in a series of pediatric kidney transplantation: predictive value of biopsy. PMID- 9745579 TI - Postransplant lymphoproliferative disease following renal transplantation: a multicenter retrospective study of 41 cases observed between 1992 and 1996. French Speaking Transplantation Workshop. PMID- 9745580 TI - Can we improve the results of simultaneous pancreas-kidney transplantation? PMID- 9745581 TI - Impact of co-infection by hepatitis B virus and hepatitis C virus in renal transplantation. PMID- 9745582 TI - Should patients with "borderline" lesions of Banff criteria be treated by renal transplantation? PMID- 9745583 TI - Mycophenolate mofetil in cyclosporine-associated nephrotoxicity. PMID- 9745584 TI - Outcome of whole liver grafts harvested and transplanted by two different teams supports a new liver procurement organization. PMID- 9745585 TI - Consequences of heart transplantation on calcium metabolism. PMID- 9745586 TI - Atrial systolic function after heart transplantation. PMID- 9745587 TI - Organ procurement by a transplant team outside of its original center: results of renal transplantation. PMID- 9745588 TI - Consequences of brain death on coronary blood flow and myocardial metabolism. PMID- 9745589 TI - Protective effects of labetalol on myocardial contractile function in brain-dead pigs. PMID- 9745590 TI - Administration of desmopressin in brain-dead donors does not modify renal function in kidney recipients. PMID- 9745591 TI - Comparison between the reactivities of fresh, preserved at 4 degrees C and cryopreserved abdominal aortae of the rabbit. PMID- 9745592 TI - Reactivity of anti-human adhesion molecules and von Willebrand factor monoclonal antibodies with pig antigens. PMID- 9745593 TI - Target cells of non-HLA antibodies in renal transplantation. PMID- 9745594 TI - Anti-endothelial and epithelial antibodies in renal transplantation. PMID- 9745595 TI - Target antigens of post-rejection non-HLA antibodies in renal transplantation. PMID- 9745596 TI - Anti-HLA class I reimmunization after one HLA semi-identical blood transfusion in non-naive patients on a waiting list for a first renal allograft. PMID- 9745597 TI - Proposal for a new classification of HLA-DR alleles based on electric charges of pockets of amino acid residues. PMID- 9745598 TI - In vitro study of alloreactivity and microchimerism after injection of dendritic cells and anti-CD4 monoclonal antibody in a combination of Lewis-Wistar Furth rats: preliminary data. PMID- 9745599 TI - CD4 lymphocytopenia in long-term renal transplant recipients. PMID- 9745600 TI - Bcl-2 and Bax expression on rat ischemic kidney. PMID- 9745601 TI - Compared effects of random and one HLA semi-identical transfusions on alloimmunization and acute rejection episodes in first renal allograft recipients. PMID- 9745602 TI - Trends in kidney transplantation in the United States. PMID- 9745603 TI - Living donors in kidney transplantation: five-year follow-up. PMID- 9745604 TI - Factors influencing the outcome of pediatric renal transplantation at a single center. PMID- 9745605 TI - Factors associated with arterial hypertension after renal transplantation. PMID- 9745606 TI - Effects of antihypertensive drugs on renal hemodynamics of kidney transplant recipients. PMID- 9745607 TI - Effectiveness of short-term OKT3 therapy in the treatment of steroid-resistant renal allograft rejection. PMID- 9745608 TI - Development of an adult liver transplant program in a public hospital in Argentina. PMID- 9745609 TI - Liver transplantation in Brazil. PMID- 9745610 TI - Simultaneous arterial and portal revascularization in liver transplantation. PMID- 9745611 TI - alpha-Tocopherol reduces hepatocyte ischemia-reperfusion injury in an isolated rat liver model. PMID- 9745612 TI - Correlation between effluent hyaluronic acid levels and early graft function in orthotopic liver transplantation. PMID- 9745613 TI - Successful treatment of graft-vs-host disease after a second liver transplant. PMID- 9745614 TI - Evaluation of the tolerance state to renal grafts by peripheral blood mononuclear cells in patients with more than eight years of graft survival. PMID- 9745615 TI - Cyclin B1 expression in response to abrogation of the radiation-induced G2/M block in HeLa cells. AB - The G2 block is a major response of cells to DNA damage and seem to be induced independently of p53 status. It is thought that the G2 block has a protective function and allows cells to repair their DNA. The molecular events involved in the formation of the G2 block therefore are of great interest. We have used pentoxifylline, a potent G2 delay abrogator, to study the expression of an essential component of the mitosis promoting complex (MPF), cyclin B1. Cyclin B1/G2 ratios are used to show that irradiation induces a decrease in cyclin B1 expression and that pentoxifylline restores cyclin B1 expression to control level. This confirms that suppression of cyclin B1 plays a role in the formation of the G2 cell cycle delay, and that elevating cyclin B1 expression is part of the mechanism of action of pentoxifylline on G2 blocked cells. PMID- 9745616 TI - Sulindac enhances cell proliferation in DMH-treated mouse colonic mucosa. AB - In a previous study we reported that the NSAID sulindac had a marked inhibitory effect on the development of colonic tumours in mice treated with the carcinogen 1,2-dimethylhydrazine (DMH). In this study we examined the effects of sulindac in respect of cell-kinetic changes in mouse colonic mucosa as determined by flash labelling with the thymidine analogue bromodeoxyuridine (BrdUrd) at varying intervals during the process of colonic carcinogenesis. We also investigated the possibility that these changes may be modulated by misoprostol a prostaglandin E1 analogue. Four groups of 36 mice each were treated for 18 weeks with the following drug/s respectively: (1) DMH; (2) DMH and sulindac; (3) DMH, sulindac and misoprostol; and (4) DMH and misoprostol. Three animals from each group were killed each week between the sixth week and the eighteenth week after the start of the experiment. A 1-h flash label technique was employed and paraffin sections of colonic mucosa were examined. For each animal a total of 50 perfect axially cut crypts were chosen and the following parameters determined: crypt length, labelling index and labelling index distribution: the data were analysed using the computer program GLIM. For each of the four groups, crypt lengths increased significantly with the duration of treatment with no significant difference between the groups. In sulindac-treated animals the labelling index for all positions increased with duration of treatment whereas for animals not treated with sulindac there was no significant difference in labelling index with respect to duration of treatment. The administration of misoprostol did not appear to significantly alter the effects of sulindac. It is postulated that the observed increase in cell proliferation could be a compensatory phenomenon occurring secondary to loss of crypt epithelial cells by apoptosis induced by sulindac. Also the finding of an increase in labelling index mediated by a chemopreventive agent indirectly questions the rationale behind the therapeutic manipulation of crypt cell proliferation in order to reduce the risk of colon cancer. PMID- 9745618 TI - A dynamic model of proliferation and differentiation in the intestinal crypt based on a hypothetical intraepithelial growth factor. AB - A widely accepted model of the temporal and spatial organization of proliferation and differentiation in intestinal epithelial is based on a cellular pedigree with all cells descending from a few active stem cells and undergoing a sequence of transitory divisions until the non-proliferating maturing cell stages develop. Model simulations have shown that such a pedigree concept can explain a large variety of data. However, so far there is neither a direct experimental proof for the existence of an intrinsic age structure in the transitory proliferative cell stages nor for the distinction between stem and transitory cells. It is our objective to suggest an alternative model which is based on evidence for intercellular communications such as might be mediated through gap junctions. We consider the diffusion of a hypothetical intraepithelial growth factor in a chain of cells which are connected via gap junctions. Individual cells can divide if a critical growth factor concentration is exceeded. Simulation studies show that the model is consistent with many observed features of the small intestinal crypt in steady state and after perturbation. PMID- 9745617 TI - Promotion-sensitive epidermal and mammary epithelial cells maintained in suspension over agarose. AB - The molecular basis of tumour promotion is still largely unknown. In in vitro model of tumour promotion, the promotion-sensitive cells are induced to grow under anchorage-independent conditions in the presence of promoting agent. The customary way of providing such conditions is to immobilize these cells in soft agar, but such cells cannot be readily recovered to study the induced biochemical and molecular events. In the present report, we analysed these events using JB6 mouse epidermal cells maintained in suspension in liquid medium over agarose. 12 O-Tetradecanoylphorbol-13-acetate (TPA) induced anchorage-independent synthesis of DNA in promotion-sensitive P+ (but not in promotion-resistant P-) JB6 cells and this TPA-induced synthesis of DNA positively correlated with TPA-induced formation of colonies in soft agar. The TPA-induced synthesis of DNA began on or shortly before 24 h after the introduction of TPA, peaked at about 48 h and then declined to the control levels over the next several days. All trans-retinoic acid and dexamethasone inhibited and calcitriol (1 alpha,25-dihydroxy-vitamin D3) synergistically stimulated this TPA-induced DNA synthesis. Western immunoblot analysis of cyclins (A, B1, D1 and E) and p27Kip1, a cyclin-dependent kinase inhibitor, indicated that TPA induced cyclin A and cyclin B1 expression in P+ (but not in P-) JB6 cells and this induction coincided in time with TPA-induced synthesis of DNA. TPA also strongly induced cyclin D1 expression in P+ (but not in P-) JB6 cells, but this induction started prior to the expression of cyclin A and cyclin B1. TPA did not affect the expression of either cyclin E or p27Kip1 to any significant extent. We also found that NMU38 rat mammary epithelial cells were operationally equivalent to the promotion-sensitive P+ JB6 cells, but in these cells 17 beta-oestradiol exerted a strong synergistic effect on TPA-induced synthesis of DNA. Based on these observations, we tentatively propose a sequence of molecular events which possibly lead to the anchorage-independent synthesis of DNA in these cells. PMID- 9745619 TI - The surgical treatment of morbid obesity. PMID- 9745620 TI - 13th International Chromosome Conference. September 8-12. Ancona, Italy. Abstracts. PMID- 9745621 TI - The plight of home health aides. PMID- 9745622 TI - Geriatrics photo quiz. Malaria, a mosquito-borne disease. PMID- 9745623 TI - Cold sore and target-like papules. PMID- 9745624 TI - Physical fitness: how to help older patients live stronger and longer. (1). AB - Most older Americans are not physically fit and do not exercise regularly. This is especially true of older women, who are weaker than men and become disabled and dependent in the later years at a much greater rate. Exercise can increase the body's metabolism and make it more efficient in burning calories. Physical fitness also makes respiration more efficient. Evidence suggests strongly that the physically fit live 2 to 3 years longer and have a better quality of life than sedentary individuals. Any time is the best time for a person of any age to start exercising. Exercise is movement--dancing, walking, lifting a weight, using the body. Older individuals tend to be willing to exercise if they are given appropriate recommendations and follow-up. PMID- 9745625 TI - Aging research: John Glenn's new mission. Interview by Robert N. Butler and Alice V. Luddington. PMID- 9745626 TI - Pharmacotherapy: strategies to control drug costs in managed care. AB - Pharmacotherapy remains one of the most cost-effective interventions physicians can provide to manage the medical conditions of older patients. Because many older Americans have multiple diseases, the practitioner's goal is to maximize appropriate drug use while avoiding duplicative or interacting medications. Medicare managed care plans seek to provide appropriate medication for the older patient at a reasonable cost through such strategies as formularies, prior authorization, generic and therapeutic substitution, and drug utilization review. Yet, like the medications themselves, these strategies require careful attention to their risks and benefits to the individual patient. PMID- 9745627 TI - Creativity and aging: ramifications for research, practice, and policy. AB - To the extent that the nature, potential, and prevalence of creativity in later life are misunderstood, the research, practice, and policies that address the needs and potential contributions of older adults will suffer. There is no denying the magnitude of disease and disability associated with aging. But what is considerably underappreciated, if not denied, is the opportunity for and frequency of creative growth and expression among aged individuals. PMID- 9745628 TI - Neurochemical changes in the aging human brain: implications for behavioral impairment and neurodegenerative disease. AB - Neurotransmission is impaired in age-related disorders, such as Alzheimer's and Parkinson's diseases, which has prompted many investigations into the neurochemistry of the aging human brain. Of all the neurotransmitter systems studied, age-related changes in parameters of the serotonergic, cholinergic, and dopaminergic systems are the most reliably measured. The association of these neurotransmitters, respectively, with mood, memory, and motor function has fueled interest in how changes in neurochemistry may contribute to age-associated behavioral changes and possibly predispose older persons to diseases of late life. The evidence suggests that impaired neurotransmission may be responsible for at least some of the behavioral abnormalities associated with aging. Moreover, age-related neurodegenerative diseases may evolve from the interaction between defects in specific neurochemical mechanisms and as-yet undefined pathophysiologic processes. PMID- 9745629 TI - Functional MRI for studying episodic memory in aging and Alzheimer's disease. AB - Anatomic and functional neuroimaging have a potential for clarifying the differential diagnosis of dementia and for evaluating new treatments for memory impairment. Our neuroimaging research examines the neural abnormalities underlying memory impairment in aging and Alzheimer's disease. We use data from high-resolution magnetic resonance imaging (MRI) to measure hippocampal volumes, and data from high-speed echo-planar imaging to evaluate cortical physiology. The results obtained with neuroimaging techniques are then related to the subject's performance on memory tasks (encoding and retrieval) performed inside the scanner during functional MRI. PMID- 9745630 TI - Motor system changes in the aging brain: what is normal and what is not. AB - Age-related changes in the nervous system may present with physical signs that are not unlike early manifestations of several clinical disorders. Gait disturbances (immobility), balance difficulties (instability), and certain motor control problems (i.e., tremor) are not necessarily signs of a disease state. The clinician needs to be reminded that most physiologic functions decline at a rate of 1% per year, beginning at age 30. Often compounding "natural" decline are the motor problems related to disuse. This is especially true for the inactive individual suffering from depression, cardiac or pulmonary insufficiency, painful joint and muscle conditions, substance abuse and, sometimes, simply social isolation. PMID- 9745631 TI - Psychotropic drugs and the aging patient. AB - Patients older than age 65 currently compose 13% of the US population, yet they receive 35% of all prescribed medications. In older patients, the complications of psychotropic drugs alone constitute a highly significant health problem. Pharmacokinetic and pharmacodynamic differences secondary to age or illness require careful consideration. Accumulation of drug might result from declining cardiovascular or renal function, alteration of body composition, or genetic or acquired inhibition of drug metabolism. As patients age, there is a general reduction in homeostatic mechanisms such as postural control, orthostatic circulatory responses, and visceral muscle function that may result in adverse drug experiences. specific receptor and neurotransmitter changes associated with senescence include reductions in central cholinergic and dopaminergic activities that lead to greater sensitivity to medications acting on these systems. Clinical vigilance is particularly important when prescribing newly available antidepressants and antipsychotics, since typically these medications are not systematically evaluated in older subjects before their release. PMID- 9745632 TI - Vitamin E and other endogenous antioxidants in the central nervous system. AB - The nervous system is protected from free-radical-induced oxidative damage by a number of antioxidants. These include small-molecular-weight substances such as vitamins C and E as well as the large protein molecules superoxide dismutase, glutathione peroxidase, and others. These antioxidants are often localized in specific portions of the cell. They also show considerable biological interactions with one another. These factors modulate their antioxidant activity. Oxidative stress has been implicated in the pathogenesis of some disorders of the brain; hence, antioxidants have become attractive therapeutic agents. Among the antioxidants, vitamin E has shown some promise in the treatment of Alzheimer's disease and cardiovascular disease. No investigations have been conducted to examine whether vitamin E or other nutritional antioxidants can be used prophylactically to prevent the incidence of degeneration or other pathologic processes in the brain. PMID- 9745633 TI - Hormones and the aging brain. AB - There is a growing appreciation of the role of ovarian hormones as modulators of neuronal function within the central nervous system. Ovarian failure has long been known to result in reversible changes in mental function, affect, and behavior. Only recently have we begun to appreciate the potential role of these hormones, specifically estrogen, in the aging of the brain and in the expression of Alzheimer's disease. As a consequence of the estrogen deficiency state of the postmenopausal woman, brain aging may be accelerated, resulting in the greater incidence of injurious falls and accidental injuries in women than in men of the same age. This sex steroid deficiency in postmenopausal women may also account for the earlier expression of Alzheimer's disease in women. PMID- 9745634 TI - Current pharmacotherapies for Alzheimer's disease. AB - This brief overview will describe some of the current anti-Alzheimer's disease (AD) agents. The relevance of the cholinergic deficit in AD is well-established. Cholinesterase inhibitor (CEI) drugs represent the only FDA-approved primary treatment options for AD as of April 1998. Modest efficacy for AD now has been shown in well-designed clinical trials for six separate CEI agents. Only two, tacrine and donepezil, are currently on the market in the United States, but several others, including rivastigmine (ENA-713), metrifonate, and physostigmine CR could be available by the end of 1998. Three other treatment strategies are being pursued. Estrogen replacement therapy as a treatment for AD in postmenopausal women is under active investigation. Analogously, clinical studies provide evidence that individuals using anti-inflammatory agents have a lower probability of developing AD. The success of alpha-tocopherol and selegiline in a recently conducted 2-year, double-blinded, placebo-controlled trial supports the hypothesis that oxidative stress plays a role in AD. PMID- 9745635 TI - Personality and psychopathology in late life. AB - Personality structure, defined as an enduring, consistent, and unique set of behavioral traits that differentiates one individual from another, forms the foundation of a person's response to both normal and pathologic aging. Although normal aging of the brain may not greatly influence personality, brain injury and disease often lead to personality change. Overall personality traits have been found to be stable within cohorts over time, although with several discrete changes. An older individual's responses to age-related stress will depend on the balance of personality strengths and weaknesses. Severe or multiple stresses in late life may overwhelm an individual's coping skills and lead to personality change. Personality disorders, defined as enduring and pervasive patterns of maladaptive behaviors, are challenging to diagnose in late life and have a variable course depending on the type of personality disorder or disorders. More research is needed to improve diagnosis and better understand the manifestations of late-life personality disorders. PMID- 9745636 TI - Sleep in aging. AB - Sleep-related complaints, particularly insomnia and wakefulness at night, are common. The causes are multiple, and include normal, if not ideal, changes in sleep stages and organization with age. The prevalence of sleep disorders is also known to increase with advancing age. These factors are even more exaggerated in the sleep of older individuals who suffer from dementia, and may account for nocturnal agitation in this group. PMID- 9745637 TI - Stroke and the aging of the brain and the arteries. AB - Stroke continues to be the third most common cause of death and a major cause of disability among those aged 70 years and older. The risk of stroke doubles for every decade after age 55. It is 25% higher in men. Age, cardiovascular disease, and hypertension are major determinants of cerebral blood flow; all have a negative impact on cerebral reperfusion. The risk of stroke can be reduced at any age by treating and correcting concomitant risk factors: hypertension; heart disease and cardiac arrhythmias (treatment with anticoagulants); transient ischemic attacks (treatment by platelet inhibitors or anticoagulants); and carotid stenosis (by endarterectomy). Cessation of smoking, control of diabetes, reduction of serum lipids, and control of obesity can reduce the risk of stroke. When stroke occurs, early treatment with rt-PA and aggressive patient care results in reduced mortality and morbidity and makes for better neurologic outcomes. Finally, prevention of stroke reduces risk of vascular dementia and makes a better functioning advanced age. PMID- 9745638 TI - Effects of chronic hypertension on cognitive functioning. AB - Arterial hypertension and blood pressure level are associated with modestly accelerated cognitive decline over the adult life span and with moderately increased risk of poor cognitive performance at all ages. Correlations between white matter lesions in brain and elevated blood pressure provide indirect evidence that structural and functional changes in brain over time may lead to lowered of cognitive functioning when blood pressure control is poor or lacking. Preventive methods designed to lower population blood pressure, early detection, and aggressive treatment of hypertension are important to prevent accelerated cognitive decline in the individual and to preserve cognitive ability in the population. Some evidence suggests that poorly controlled hypertension may predispose to the dementias, but more work is needed to ascertain this. PMID- 9745639 TI - Driving is transportation for most older adults. AB - The number of people in the United States older than age 65 is expected to double to 60,000,000 within the next 25 years. It is likely that in the future more older persons will drive longer into their lifetimes than they do now. Actions taken now to prepare for this population expansion will have a profound effect on the lives of future older Americans. Functional disabilities among some older people may make it difficult for them to continue to be safely mobile. A crucial issue is to determine who can drive safely and who cannot. A corollary issue is how to rehabilitate those whose functional abilities can be enhanced. For those who cannot be rehabilitated, decisions need to be made on either modifying or stopping driving, and we must facilitate the use of other options that allow older people to remain mobile. The National Highway Traffic Safety Administration and other public and private organizations are now developing programs to enable the public and private sectors to address these issues. PMID- 9745640 TI - A biopsychosocial perspective on behavior problems in Alzheimer's disease. AB - The challenges of managing behavior problems in persons with dementia require new models that support interdisciplinary approaches. The Ecosystemic Biopsychosocial Grid is presented as a systems approach to assessing and organizing care. It assesses both resources and barriers across 10 realms of daily experience and identifies approaches that can maximize the balance between the capacities of persons with dementia and the demands placed upon them by their environment. Issues specific to Alzheimer's disease are reviewed across the biopsychosocial continuum. PMID- 9745641 TI - Adapting living environments for persons with Alzheimer's disease. AB - Skilled nursing facilities, assisted living facilities, and specialized housing can be enhanced by incorporating dementia-oriented adaptations that better match the environment to the capabilities of individuals at different stages of dementia. Environmental adaptations can be made through architectural, interior, and landscape design specifications. New models of care for Alzheimer's disease are emerging all along the long-term care continuum. This trend has been accompanied by a gradual growth in the knowledge base about designing and delivering specialized dementia care. Three components of care are critical: (1) environmental design, (2) program development, and (3) staff development. Six different models of dementia care were identified based on an analysis of care attributes that represent the critical components of care. These models differed in terms of caregiver satisfaction and resident characteristics (ADL scores, cognition, stage of dementia, agitation, and antipsychotic drug use). PMID- 9745642 TI - Lovers, loners, and lifers: sexuality and the older adult. AB - With the aging of the baby boomers, increased attention is being given to sexuality and aging. This article discusses what is found in the current literature regarding the sociocultural, psychological, and physical factors affecting sexuality in aging individuals. PMID- 9745644 TI - The aging self in an 'ageist' culture: getting beyond denial. AB - What is certain is that we will age. Demonizing this inevitability hardly encourages people to get on with the distinctive work often possible for the first time only in life's later years. That work calls for the belated recognition and some integration of humanity's enduring ambivalences: we are both angels and animals, both natural creatures and societally nurtured. PMID- 9745643 TI - The vegetative and minimally conscious states: ethical implications. AB - Modern medical technology has created new syndromes of severe and permanent brain damage. In the first 25 years of the right-to-die debate, the permanent vegetative state has been the paradigmatic neurologic syndrome for decisions to discontinue treatment. In the near future, however, a far more problematic syndrome may be even more important in the right-to-die debate, the minimally conscious state. This paper presents a few of the medical and ethical similarities and differences between the permanent vegetative and minimally conscious states and discusses how value-laden these decisions may become in the future. PMID- 9745645 TI - An investigation of dental student practice preferences. AB - This study explored characteristics hypothesized to influence dental student long range practice preferences. Dental students (N = 264) for this study were selected at random from all dental students in one state who rated practice arrangements they most prefer to be in five years after graduation. The rating instrument was composed of two items measured on 5-point Likert-type scales. The independent variables were student gender, year of dental study, and practice arrangement. The research design was a 2 x 4 x 3 completely randomized, fixed factor, factorial analysis of variance (ANOVA). The ANOVA model explained 48 percent of the variance in student practice arrangement ratings. Both males and females rated solo ownership as the most favorable practice arrangement. Males rated the solo owner arrangement more favorably than did females, while females had a stronger preference for the employee practice arrangement than did males. Study results have implications for dental educators in their efforts to structure curricula to meet diverse student needs, to assess personnel needs and enhance the accuracy of projections, and to evaluate the effect of practice preferences on the health care system. PMID- 9745646 TI - Survey of the information-seeking patterns of dental hygienists. AB - This descriptive study explored the methods that dental hygienists in northern British Columbia have utilized to access information. A self-administered questionnaire sent to 130 dental hygienists registered in that geographic experienced a response rate of 81.5 percent. The respondents preferred and utilized traditional information sources such as discussions with colleagues, journal articles, and mailings from professional associations and licensing bodies. The least utilized information sources were the indices to the literature and electronic information sources. Geographic isolation, lack of electronic information sources, and costs were identified as the top three barriers to information access. Dental hygienists may need to acquire or improve computer literacy skills while in school and through continuing education to enable them to use the newer methods of electronic information retrieval and communications because dental hygienists need to access a variety of information sources to provide quality care. PMID- 9745648 TI - After-hours emergency policies at U.S. dental schools. PMID- 9745647 TI - Radiographic examination of dental school comprehensive care patients. PMID- 9745649 TI - Comparing traditional lecture vs. computer-based instruction for oral anatomy. PMID- 9745650 TI - Clinical research, dental education, and the NIH Clinical Research Training Program. PMID- 9745652 TI - The termination package that just didn't register. PMID- 9745655 TI - The road to Hana. PMID- 9745651 TI - Linking postdoctoral general dentistry programs with managed care programs and settings. AB - This report presents the results of a small workgroup convened by the American Association of Dental Schools to examine experiences related to postdoctoral general dentistry programs linked with managed care systems and clinical settings. The workgroup was a component of an Association effort to identify and promote innovative and nontraditional methods by which the number of postdoctoral general dentistry (PGD) positions can be increased to meet current demand for PGD education. The participants identified factors and conditions that they believed to be critical to the planning, development, and conduct of PGD programs with substantial linkages with managed care systems and settings. The information should be helpful to others as they consider opportunities to establish PGD programs or increase their number of PGD training positions. PMID- 9745656 TI - Mentoring a novice chief nurse executive. PMID- 9745657 TI - How nurses become leaders. Perceptions and beliefs about leadership development. AB - Creating a professional work environment where nurses can develop the skills and expertise needed to be successful leaders is one strategy that can help ensure successful nursing leaders for the future. Understanding which factors influence leadership development is essential to create such an environment. The author considers what those factors may be and how they interact to facilitate the development of leadership skills and expertise among nurses. PMID- 9745658 TI - Measuring and evaluating hospital restructuring efforts. Eighteen-month follow-up and extension to critical care, Part 1. AB - Increasingly, hospital restructuring is viewed with skepticism because of a lack of systematic and rigorous evaluation of its impact on quality of care. This first article in a two-part series describes comprehensive evaluation of the effects of hospital restructuring on patient satisfaction, nurse satisfaction, costs of care, and clinical quality on four medical-surgical units at a large tertiary hospital. In addition, early application of the model to critical care is described. A quasiexperimental pre- and post-design combined with concurrent control units for selected measures was the overall strategy. The authors conclude that comprehensive restructuring of hospital-based care can take place in a manner that preserves multiple dimensions of quality while decreasing costs. This only can be ascertained, however, through rigorous and systematic measurement and evaluation. Part 2 will detail application and evaluation of the restructuring model in the critical care environment. PMID- 9745659 TI - The influence of structure, staff type, and managed-care indicators on registered nurse staffing. AB - Nurse administrators need methods to evaluate and compare staffing across a variety of hospitals because the degree of reengineering that actually has occurred in their communities can be difficult to assess. Multivariate analysis of factors affecting hospital registered nurse (RN) staffing in western New York revealed that the significant factors were the type of unit, nursing model, rural location, and use of aides and unit secretaries. Managed-care factors and alternative uses of staff did not affect RN staffing. Regional market variations may have significant impact on staffing solutions adopted by nurse executives. PMID- 9745660 TI - Interdisciplinary collaboration and discharge planning communication for elders. AB - The effects of personal characteristics and perceptions of interdisciplinary collaboration on discharge planning communication were examined for nurses, physicians, and social workers in two hospitals. The model for the study explained 61.7% of the variance in discharge planning communication for nurses. For all 142 health professionals, communication openness with social workers, problem solving between nurses and physicians, and collaboration with social workers were important to discharge planning communication. For nurses, communication satisfaction with patients and families also was important. PMID- 9745661 TI - Hospital ethics committees and nurses' participation. AB - The hospital ethics committee's mandates of patient care review, policy formation, and education make them central to nurses and healthcare delivery. In a study examining nurses' communication exchange frequency and perceived effectiveness as members of hospital ethics committees, nurses represented the largest proportion, were moderately active, and rated their participation effectiveness the highest: they are more involved in discussions regarding patients than policy formation and education. Nurse administrators can provide strategies and education for nurses that enhance nurses' participation in all three committee functions. Policies affect patient care; therefore, policy decisions, too, benefit from nurses' participation. PMID- 9745662 TI - Reduced bovine leukaemia virus proviral load in genetically resistant cattle. AB - The bovine leukaemia virus (BLV) is an exogenous retrovirus that is closely related to the human T cell leukaemia viruses. Genetic resistance and susceptibility to persistent lymphocytosis (PL), an advanced subclinical stage of infection characterized by a polyclonal expansion of the infected B cell population, have been mapped to structural motifs in bovine MHC DRB3 (class II) alleles. To determine whether alleles of DRB3 influence the number of BLV infected B cells in peripheral blood, seven pairs of Holstein cows naturally infected with BLV were matched on the basis of DRB3 genotype (resistance or susceptibility to PL), age, and year of seroconversion. Flow cytometry was used to separate B cell populations that then were tested for the presence of provirus by a single-cell PCR methodology. Animals with the PL-resistance associated DRB3.2*11 allele had significantly fewer BLV-infected B cells than did age- and seroconversion-matched cows with DRB3 alleles associated with susceptibility to PL. Our results demonstrate that DRB3 or a closely linked gene may play a direct role in controlling the number of BLV-infected peripheral B cells in vivo. Association of MHC class II alleles with resistance to disease progression in cattle naturally infected with BLV provides a unique immunogenetic model for the study of host resistance to human and other animal retroviral infections. PMID- 9745663 TI - Genetic diversity of Asian water buffalo (Bubalus bubalis): mitochondrial DNA D loop and cytochrome b sequence variation. AB - Swamp and river buffalo mitochondrial DNA (mtDNA) was sequenced for 303 bp of the cytochrome b gene for 54 animals from 14 populations, and for 158 bp of the D loop region for 80 animals from 11 populations. Only one cytochrome b haplotype was found in river buffalo. Of the four haplotypes identified in swamp buffalo, one found in all populations is apparently ancestral both to the other swamp haplotypes and to the river haplotype. The phylogenetic relationships among the 33 D-loop haplotypes, with a cluster of 11 found in swamp buffalo only, also support the evolution of domesticated swamp and river buffalo from an ancestral swamp-like animal, most likely represented today by the wild Asian buffalo (Bubalus arnee). The time of divergence of the swamp and river types, estimated from the D-loop data, is 28,000 to 87,000 years ago. We hypothesise that the species originated in mainland south-east Asia, and that it spread north to China and west to the Indian subcontinent, where the rive type evolved and was domesticated. Following domestication in China, the domesticated swamp buffalo spread through two separate routes, through Taiwan and the Philippines to the eastern islands of Borneo and Sulawesi, and south through mainland south-east Asia and then to the western islands of Indonesia. PMID- 9745665 TI - Genetic relationships among European cattle breeds. AB - Genetic relationships among 37 European cattle breeds were investigated using blood group and serum protein polymorphisms. The 18,859 animals included in the study represented a random sample from pedigree populations in the UK. Within breed variation was estimated by average heterozygosity and number of alleles observed, and breed relationships were evaluated by genetic distance. Standard errors of the heterozygosity, number of alleles and genetic distance were obtained by bootstrapping. The significance of breed differences was tested using an exact test of differentiation. French, Italian and Channel Island breeds were found to have generally higher heterozygosities and a greater number of alleles than breeds from mainland Britain and North Europe. Genetic distances ranged between 0.011 (+/- 0.005) and 0.309 (+/- 0.071). Two major breed groups were identified; a group of French, Italian and Channel Island breeds together with the Simmental and Gelbvieh, and a second group consisting of the mainland British and North European breeds. The exact test of breed differentiation showed all breeds to be significantly different from one another (P < 0.0001). Overall relationships among breeds reflected their geographical origin and common ancestry rather than the agricultural use for which the breeds have been selected. PMID- 9745664 TI - Comparative mapping of bovine chromosome 13 by fluorescence in situ hybridization. AB - We present chromosomal fluorescence in situ hybridization (FISH) results that both extend the HSA20/BTA13 comparative map as well as cytogenetically anchor two microsatellite markers. A bovine bacterial artificial chromosome (BAC) library was screened for conserved genes (type 1 loci) previously assigned to HSA10 or HSA20 and BTA13, and for microsatellites selected from two published BTA13 linkage maps. Clones from six out of nine comparative loci and both microsatellites were found represented in the BAC library. These BAC clones were used as probes in single colour FISH to determine the chromosome band position of each locus. As predicted by the human/bovine comparative map, all type 1 loci mapped to BTA13. Because single colour FISH analysis revealed that the loci were clustered within the distal half of BTA13, dual colour FISH was used to confirm the locus order. Established order was centromere-PRNP-(SOD1L/AVP/OXT) (BL42/GNAS1)- HCK-CSSM30. The findings confirm the presence of a conserved HSA20 homologous synteny group on BTA13 distal of a HSA10 homologous segment. PMID- 9745666 TI - Characterization of beta-defensin prepropeptide mRNA from chicken and turkey bone marrow. AB - Four avian beta-defension prepropeptide cDNA sequences [gallinacins: Gal 1 (synonym CHP 1, chicken heterophil peptide 1), and Gal 2; turkey heterophil peptides: THP 1 and THP 2] were amplified from chicken or turkey bone marrow mRNA samples, respectively. Partial chicken beta-defensin cDNA sequences were obtained using degenerate primers based on chicken peptide sequences (Gal 1/CHP 1 and Gal 2). The complete cDNA sequences of the chicken beta-defensins were then determined by designing specific intrapeptidal primers, from the newly acquired sequence, and pairing one primer with a specific poly A primer tail sequence (3' end) and the other primer with an adapter primer in a 5' rapid amplification of cDNA ends (RACE) reaction. The two, turkey beta-defensins were amplified from turkey marrow using primers designed from chicken beta-defensin preproregions. The complete amino acid sequences for the prepropeptides were deduced for all four avian beta-defensins. Previously, only partial mature peptide sequences for the turkey beta-defensins and complete mature peptide sequences for the chicken beta-defensins were known. All sequences obtained translated accurately to complete and partial amino acid sequences reported for beta-defensins purified from chicken and turkey heterophil granules except for one additional amino acid for Gal 1/CHP 1. The four deduced beta-defensin proregions lack the long, negatively charged propiece reported in classical defensin proregions. These regions are thought to stabilize and inactivate the positively charged mature peptide and target the propeptide to the storage granule. Instead, these beta defensin proregions are shorter and similar to storage granule-free beta defensins proregions reported for bovine tracheal antimicrobial peptide (TAP) and lingual antimicrobial peptide (LAP). These are the first prepropeptide beta defensins from leukocyte granules to be completely characterized. PMID- 9745667 TI - Parameters of the chicken genome (Gallus gallus). AB - As more information on the chicken genome is gathered, it is becoming increasingly more important to be able to correlate genetic and physical maps. Quantitation of the chicken karyotype is important in establishing parameters which define the genome. Here we report on the physical lengths of the chicken macrochromosomes and establish the DNA content of each, thus identifying implicitly how much of the genome is represented by the microchromosomal component. For the first time, genetic and physical data on the chicken karyotype are presented in relation to one another. PMID- 9745668 TI - Construction and characterisation of a gridded chicken cosmid library with four fold genomic coverage. AB - Gridded genomic libraries are crucial for the positional candidate gene approach. For this purpose we constructed a gridded genomic library from a female chicken using the vector sCos 1. About 110,000 cosmid clones were grown and replicated in 384-well plates. An average insert size of 39 kb was calculated from the analysis of 68 randomly selected clones. No chimerism could be observed from 31 in situ hybridisations. One replica of the library (number 125) has been transferred to the Resource Centre/Primary Database (RZPD) of the German Human Genome Project (DHGP). The whole library was gridded onto four nylon filters at high density for efficient identification of cosmid clones by colony hybridisation. Twenty-two probes were used for screening the library and each of them gave at least one positive signal. This result is in good agreement with a four-fold coverage of the genome as estimated from the insert length and number of recombinant clones. This library provides a powerful tool for rapid physical mapping and complex analysis of the chicken genome. PMID- 9745669 TI - Characterisation of a bovine/murine hybrid cell panel informative for all bovine autosomes. AB - A bovine/murine hybrid cell panel consisting of 57 cell lines was typed with 124 markers by PCR. Southern hybridisation and isozyme analysis in order to establish its utility as a resource for genome mapping. All bovine chromosomes, including the sex chromosomes were represented in the panel. Computerised analysis of syntenies indicated that there are no cell lines containing only a single bovine chromosome. The panel was used to map 10 new bovine microsatellite markers, and the MYL6 and CPE genes. This panel is informative for all bovine chromosomes other than the sex-specific region of the X chromosome and can be used in synteny mapping studies. At present, due to the relatively small number of markers typed, the resolution of the panel does not go beyond the chromosomal level. PMID- 9745670 TI - Bovine microsatellite loci are highly conserved in red deer (Cervus elaphus), sika deer (Cervus nippon) and Soay sheep (Ovis aries). AB - We tested 174 bovine microsatellite primer pairs for use in a primitive breed of sheep and two species of deer. Of 173 markers, 127 (73.4%) gave a product in Soay sheep (Ovis aries) of which 54 (42.5%) were polymorphic. One hundred and twenty nine of 174 (74.1%) markers gave a product in red deer (Cervus elaphus) of which 72 (55.8%) were polymorphic. In sika deer (Cervus nippon) 126 of 171 (73.7%) microsatellite primers gave a product with 47 (37.3%) polymorphic. The proportion of bovine microsatellite loci conserved across artiodactyl species was significantly greater in this study than previously reported. Reasons for this high degree of microsatellite conservation are discussed. We suggest that a high resolution comparative map of the artiodactyls can be constructed using microsatellites. PMID- 9745671 TI - Evaluation of ROM1 as a candidate gene in generalised progressive retinal atrophy in dogs. AB - Generalised progressive retinal atrophy (gPRA) is a heterogeneous group of hereditary diseases causing degeneration of the retina in dogs and other animals. The genetic origin is unknown in most cases. We have screened the coding sequence of the ROM1 gene for disease causing mutations in Tibetan Terriers, Miniature Poodles, Dachshunds and Chesapeake Bay Retrievers by single strand conformation polymorphism analysis (SSCP). Two polymorphisms have been identified by sequencing, one in exon 1 in all examined breeds (position 210: G-->A; Gly40Arg and position 252: G-->T; Ala53-Ser). Another polymorphism was present in exon 2 (position 1150: C-->T and position 1195: C-->T) segregating in Miniature Poodles. None of these polymorphisms were cosegregating with gPRA rendering a disease causing mutation in the ROM1 gene unlikely. PMID- 9745672 TI - Physical ordering of six YACs from the RN region in pigs. AB - Six YAC clones representing five microsatellite markers from the RN region were mapped by fluorescent in situ hybridization (FISH) on pig metaphase chromosomes and their relative order was determined by pairwise multicolour FISH. Two of the microsatellites viz., Sw120 and Sw936 flank RN as well as the remaining three microsatellites Sw1683, Sw2083 and Sw1309. The results assigned the RN locus to the distal part of the 15q25 band. The linear order of the microsatellites was compared with the available linkage mapping data. PMID- 9745673 TI - A polymorphic (TG)n microsatellite in an intron of the canine tyrosine transaminase gene. PMID- 9745674 TI - A length polymorphism in an intron of the canine polycystic kidney disease 1 gene. PMID- 9745675 TI - KRN1 maps to bovine chromosome 29. PMID- 9745676 TI - Dinucleotide repeat polymorphisms in waterfowl (family Anatidae): characterization of a sex-linked (Z-specific) and 14 autosomal loci. PMID- 9745677 TI - A PCR-RSP Csp6I site in the canine ornithine aminotransferase (OAT) gene. PMID- 9745678 TI - Assignment of the canine microsatellite ZuBeCa3 to canine chromosome 9q21-q22. PMID- 9745679 TI - Length polymorphism in a CT-rich microsatellite in an intron of the canine tyrosinase-related protein-2 gene. PMID- 9745680 TI - A BsII PCR/RFLP in the renin binding protein (RnBP) gene on canine chromosome X. PMID- 9745681 TI - A polymorphic (CA)n microsatellite in the canine lecithin:cholesterol acyltransferase gene. PMID- 9745682 TI - A BseRI PCR/RFLP in an intron of the canine phenol sulfotransferase gene. PMID- 9745683 TI - The highly polymorphic canine microsatellite ZuBeCa2 is localized on canine chromosome 1q210-q211. PMID- 9745684 TI - A NlaIII PCR/RFLP in an intron of the retinitis pigmentosa GTPase regulator gene (RPGR) on the canine X chromosome. PMID- 9745685 TI - Ostrich microsatellite polymorphisms at the VIAS-OS4, VIAS-OS8, VIAS-OS14, VIAS OS22, and VIAS-OS29 loci. PMID- 9745686 TI - Detection of an SSCP within intronic sequence of the bovine TIMP-2 gene. PMID- 9745687 TI - The gallium-deferoxamine complex: stability with different deferoxamine concentrations and incubation conditions. AB - Previous studies report that deferoxamine (DFO) binds metallic ions such as Fe3+, In3+ and Ga3+ with very high affinity. This property of DFO has been utilized to label DFO-coupled compounds with radiometals such as 67Ga and 111In. We have studied the effect of low DFO concentrations and of different incubation conditions on the stability of the 67Ga-DFO complex. In our experience high (> 5 microM) DFO concentration appears to be critical in obtaining high radiochemical purity of such complexes. PMID- 9745688 TI - Beta-decay half-lives and level ordering of 102m,gRh. AB - Beta-decay half-lives of the ground state and an isomer of 102Rh have been determined 207.3(17) d and 3,742(10) y, respectively, by gamma-ray decay curves following each beta-decay. It has been found that a state (2-) which has a shorter half-life (207.3 d) is the ground state from the result that the half life of the 41.9 keV isomeric gamma-transition was equal to 3.742 y. It has also been confirmed that the 41.9 keV transition is certainly an isomeric transition with X-gamma coincidence measurement. PMID- 9745689 TI - Determination of bone mineral density in the third lumbar vertebral body using photon absorptiometry techniques. AB - Dual-photon absorptiometry and triple-energy X-ray absorptiometry were used to investigate the total bone mineral content and density as well as the trabecular bone mineral density in the third lumbar vertebral body. Both anteroposterior (AP) and lateral (LAT) measurements were performed. By combining the two projections it was found that the mean trabecular bone mineral density for all 202 subjects included in the study was 52% (SD +/- 20%) of the total bone mineral density in the third lumbar vertebral body. The mean trabecular bone mineral density as a fraction of the total vertebral body bone mineral density decreased as a function of age. The relative annual change in this fraction differed between males and females. It was also found that neither trabecular nor total bone mineral density differed significantly between male and female subjects aged 25-35 years, and bone mineral density (BMD), expressed in g/cm3, showed no correlation to subject height, body weight or body mass index (BMI). Male and female individuals showed different rates of change of trabecular bone mineral density with age. PMID- 9745690 TI - Radiochemical purification of no-carrier-added scandium-47 for radioimmunotherapy. AB - A procedure, adaptable to large-scale remote operation, was developed to purify no-carrier-added (NCA) 47Sc from irradiated Ti targets. Methods based on extraction chromatography and cation-exchange chromatography were compared. Results of this comparison led to the development of an optimized procedure based on cation-exchange with Dowex AG 50W-X4 and 47Sc elution with HCl/HF. This method gave 90-97% overall 47Sc recovery, with a Ti separation factor greater than 2.4 x 10(-5), and specific activities > or = 0.9 GBq microgram-1. Use of the 47Sc product, for labeling monoclonal antibodies, resulted in consistent labeling yields of > or = 90%. PMID- 9745691 TI - Synthesis of a F-18 labeled analog of antitumor prostaglandin delta 7-PGA1 methyl ester using p-[18F]fluorobenzylamine. AB - A fluorine-18 labeled analog of an antitumor prostaglandin delta 7-PGA1 methyl ester, 15-deoxy-13,14-dihydro-delta 7-PGA1 4-[18F]fluorobenzyl amide ([18F]3), was synthesized as a tracer candidate for detecting tumors with positron emission tomography. p-[18F]Fluorobenzylamine (p-[18F]FBnA) used as a labeled precursor for the synthesis of [18F]3 was prepared by fluorination of a 4-N, N, N trimethylammonium-benzonitrile triflate with [18F]fluoride and subsequent reduction with borane-dimethylsulfide. Radiochemical yield and purity of p [18F]FBnA obtained were 39-49% (decay uncorrected) and 91-96%, respectively, after C18 Sep-Pak purification. Treatment of p-[18F]FBnA with a 15-deoxy-13,14 dihydro-delta 7-PGA1 N-succinimidyl ester in acetonitrile and subsequent HPLC purification gave radiochemically pure (> 99%) [18F]3 with a 58% decay uncorrected yield. The total synthesis time was 70 min from the start of the radiosynthesis of p-[18F]FBnA. PMID- 9745692 TI - The synthesis, radioiodination and preliminary biological study of the new carboxylic derivatives of dithizone. AB - Synthesis, characteristics and radioiodination of the new carboxylic derivatives of dithizone are described in this paper. We have applied the carboxy dithizones for preparation of radioactive compounds by coupling with [131I]-histamine. Preliminary biological studies of the new radiodithizone were done in rats after two different application routs: peripheral i.v. injection and direct injection to splenic artery. Biodistribution of the carboxy dithizone-[131I]-histamine conjugate (i.v. injection) was quite different than that for free [131I] histamine. However, uptake of activity in pancreas was low (0.81% g-1 of tissue). Direct application of the conjugate to splenic artery resulted in high activity retention in pancreas after 30 and 45 min post injection (respectively 8.8 and 12.4% g-1 of tissue) indicating potential usefulness of the new radiodithizone for in vivo monitoring of pancreas. PMID- 9745693 TI - Synthesis of 2-iodo- and 2-phenyl-[11C]melatonin: potential PET tracers for melatonin binding sites. AB - Two 11C-labelled melatonin derivatives, 2-iodo-[11C]melatonin (2-iodo-5-methoxy N[11C-acetyl]-tryptamine, an agonist) and 2-phenyl-[11C]melatonin (2-phenyl-5 methoxy-N[11C-acetyl]tryptamine, a putative antagonist) were synthesized from [11C]carbon dioxide. The reaction sequence was common to both compounds and consisted of three steps: (i) carbonylation of methyl magnesium bromide with [11C]carbon dioxide, (ii) conversion of the adduct to [11C]acetyl chloride, (iii) acetylation of the amine precursors (2-iodo-5-methoxy-tryptamine or 2-phenyl-5 methoxy-tryptamine) with [11C]acetyl chloride. The precursors were especially prepared. The radiochemical yield was 19% for 2-iodomelatonin and 32% for 2 phenymelatonin, based on [11C]carbon dioxide; the specific activity ranged from 300 to 600 mCi/mumol. Both labelled 2-substituted-melatonins are intended to be used as radiotracers to study melatonin binding sites in man with positron emission tomography. PMID- 9745694 TI - Stabilization of Tc-99m D,L-HMPAO preparations as a leucocyte labelling agent. AB - An attempt was made to use stabilized Tc-99m D,L-HMPAO (S-HMPAO) to label leucocytes. The radiochemical purity of Tc-99m D,L-HMPAO, labelling efficiency of leucocytes, cell viability of labelled leucocytes, and stability of S-HMPAO labelled leucocytes were calculated. In comparison with commercial Tc-99m D,L HMPAO (C-HMPAO) without stabilization, immediately, at 0.5, 1, 2, 4, and 6 h after HMPAO preparation, the radiochemical purity of S-HMPAO and the labelling efficiencies of S-HMPAO labelled leucocytes were higher. S-HMPAO is more stable than C-HMPAO and can provide higher leucocyte labelling efficiency. S-HMPAO, therefore, has the potential to replace C-HMPAO as a leucocyte-labelling agent. PMID- 9745695 TI - Measurement of equilibrium factor and unattached fraction of radon progeny in Kaohsiung, Taiwan. AB - To reasonably estimate the population dose from radon, equilibrium factor, Fp, and unattached fraction, fp, of radon progeny are important parameters. A Lucas cell and a working level monitor was used for most Fp measurement. The unattached fraction was measured by SARAD EQF3020. A detached house was chosen to measure the Fp in different rooms. The Fp value depended mainly on the ventilation rate and surface to volume ratio of the rooms. Fp and fp were measured in bedrooms of 14 other dwellings. Two hospitals were also chosen for measurement of Fp in the working place using rooms located in the basement. The average Fp for dwellings was about 0.5 and the average unattached fraction was about 0.055. The radon levels in the hospitals were higher than those in the dwellings but the equilibrium factors in the hospital were very low (about 0.06). The low Fp was attributed to the use of a dehumidifier in the hospitals. Dehumidifiers are popular for reducing fungi problem induced by the high humidity in Taiwan. PMID- 9745697 TI - X-ray fluorescence analysis of bromine for the estimation of extracellular water. AB - Three methods for extracellular water (ECW) estimation via analysis of the corrected bromine space (CBS) are presented. The methods are based on the application of stable bromine as an indicator and X-ray fluorescence analysis (XRF). The content of stable bromine can be determined in samples of ulna vein blood and daily urine (the first method) as well as in microsamples of finger blood (the second method) and mixed non-stimulated saliva (the third method). The precision and accuracy of the methods were compared using the results of both repeated analysis and CBS estimation by different procedures including the routine method with 82Br as indicator. A device for XRF with a Si(Li) detector and 109Cd sources was developed for bromine determination in biological fluids. PMID- 9745696 TI - Preliminary study of dose equivalent evaluation for residents in radioactivity contaminated rebar buildings. AB - It has recently been found that several resident and office buildings in Taiwan were constructed with 60Co-contaminated reinforcing steel bar (rebar). Both governmental officials and the residents of such buildings have been concerned about this finding. In order to respond to the situation, the government has adopted a number of remedial measures, including full-scale radiation survey, dose evaluation and physical examinations of residents. This article presents three methods for evaluating the dose equivalents of the residents living in the contaminated rebar buildings by means of gamma-ray survey, necklace-type thermoluminescence dosimeters (TLDs) and the human lymphocyte chromosome aberration analyses. The results reveal that the dose evaluation by gamma-ray survey is rather conservative. Generally for the residents whose annual dose equivalents are greater than 5 mSv (0.5 rem) by gamma-ray survey, the dose equivalents from necklace-type TLDs are only within the range of 20 to 50% of the evaluated values mentioned above. For chromosome analyses, at least 500 lymphocyte cells were scored and analyzed for each resident. Most of the chromosome analysis data show that the dose equivalents received by residents are lower than the detection limit of the method (100 mSv) and quite different from the estimated dose obtained from either gamma-ray survey or necklace-type TLD measurements. PMID- 9745698 TI - Radiostrontium analytical method using crown-ether compound and Cerenkov counting and its applications in environmental monitoring. AB - The radiostrontium content in environmental samples was determined by chemical analysis by means of the fuming nitric acid method and ion exchange method with low-level beta counting and the newly developed method using crown-ether compound. Counting was performed with a low-background counter and a liquid scintillation counter together; the latter was the Cerenkov counting method. All results obtained by these three methods were in good agreement. The time for chemical separation of radiostrontium, using crown-ether compound, is much faster than fuming nitric acid and ion exchange methods. However, due to the high background of the liquid scintillation counter, the detection limit for Cerenkov counting is about two times higher than that for low-background counting. PMID- 9745699 TI - Electron density of normal and pathological breast tissues using a Compton scattering technique. AB - Compton (incoherently) scattered photons which are directly proportional to the electron density of the scatterer, have been employed in characterising human breast tissues. Gamma ray photons scattered incoherently from normal and pathological breast tissue samples of nine breast cancer patients were measured using a high purity germanium detector and an americium (Am-241) source. The breast tissue samples were obtained from female patients undergoing mastectomy. The samples were examined in freeze dried form and the results were corrected for the reduction in the water content of each tissue type by use of the Mixture Rule. This study is aimed at providing electron density information in support of the introduction of new tissue substitute materials for mammography phantoms. PMID- 9745700 TI - Assessment of the transfer of 137Cs in three types of vegetables consumed in Hong Kong. AB - A dynamic food chain model has been built for the modeling of the transfer of 137Cs in three types of vegetables consumed in Hong Kong, namely, white flowering cabbage (Brassica chinensis), head lettuce (Lactuca sativa) and celery (Apium graveolens). Some parameters have been estimated from the experimental data obtained in this work. The experimental data include the transfer factors of 137Cs from soil to the different vegetable species which are determined through high resolution gamma spectrometry, maximum crop biomasses for the vegetable species, the dry-to-fresh ratios for the vegetable species, the bulk density of soil layers and the average concentration of 137Cs in air. The derived parameters include the deposition rate and the root uptake rate, information for tillage, the logistic growth model and radionuclide concentrations in vegetables. The dynamic food chain model is solved by the Birchall-James algorithm to give the 137Cs concentration in subsurface soil, from the 0.1-25 cm soil layer, and the 137Cs concentration in harvested and unwashed vegetables. As validation of the model and parameters, the concentrations obtained experimentally and from the model are compared and are found to be in good agreement. PMID- 9745701 TI - Radionuclide accumulation in near-shore sediments along the Bulgarian Black Sea Coast. AB - The accumulation of radionuclides in Black Sea marine ecosystems was investigated by low level gamma spectrometry. Artificial as well as natural radionuclides were determined in bottom sediments samples from 35 reference locations along the Bulgarian Black Sea coast, evenly distributed from the Rumanian to the Turkish border including the main Black Sea resorts and rivers. The measurement of radionuclides in sea bed sediments was carried out during six consecutive seasons using a HPGe detector. The data obtained show that the nuclide concentrations depend strongly on the sediment nature. Results for sandy sediments are within close range, while those for slime and silt vary to a much greater extent. The radionuclide content in the sandy sediments of the main Black Sea resorts is at the lowest limit of the determined values. Small seasonal changes of radionuclide concentration in sandy sediments were observed while greater variations in slime and silt occur. From the data obtained 134Cs/137Cs and 137Csmeas/137CsChern ratios are calculated to determine the Chernobyl part of the measured 137Cs. The activities determined in the sediments for natural radionuclides correspond to those cited in the literature for natural levels, showing no additional anthropogenic contamination. A data base for the nuclide concentration values was created which will enable the modeling of radionuclide transfers by estimation of their concentration variations, accumulation and influence on the marine ecosystems. PMID- 9745704 TI - Microbial hexachlorobenzene dechlorination under three reducing conditions. AB - The potential dechlorination of hexachlorobenzene (HCB) in medium by 1,2,3 trichlorobenzene (TCB)-adapted mixed culture under three reducing conditions was investigated. It was found that strongest to weakest HCB dechlorination occurred in the order of methanogenic conditions > sulfate-reducing conditions > denitrifying conditions. Under denitrifying conditions, no dechlorination was observed during the first 20 days of incubation. Biotransformation occurred in this order: HCB-->pentachlorobenzene (PCB)-->1,2,3,5-tetrachlorobenzene (TeCB)- >1,3,5-TCB + 1,2,4-TCB-->1,3-dichlorobenzene (DCB), HCB dechlorination was delayed following treatment with ferric chloride and manganese dioxide, but enhanced by the addition of lactate and pyruvate under methanogenic or sulfate reducing conditions, the addition of acetate had no significant effect on HCB dechlorination under any of the three reducing conditions. Sequential dechlorination was observed at concentrations of 2-50 mg/L, but at a significantly slower rate at the highest concentrations. PMID- 9745702 TI - Analysis of airborne radon in an ultra-low background experiment. AB - The contribution of 222Rn to the background in a low background experiment with a germanium detector has been estimated. We have also checked the efficacy of a standard radon cleaning system. The cleaning reduces the radon concentration two orders of magnitude with respect to the air in the laboratory. The residual 222Rn represents at most 12.5% of the background in the low energy region, a value low enough for the purpose of our experiment. A detailed study of the radioactive background is presented. PMID- 9745705 TI - Bioaccumulation of musk xylene (MX) in developing and adult rats of both sexes. AB - Bioaccumulation of musk xylene (MX) was measured by GC-ECD in adult and developing Long Evans rats. Males and females were fed with MX-containing chow (0.001, 0.01, 0.033, 0.1 g MX/kg food pellets) for 10 weeks before mating. Treatment continued during pregnancy and lactation. Offspring exhibited dose dependent MX accumulation with 1/2-3/4 of adult female or 3-4 times adult male body fat levels (at 0.1 mg/kg food) at days 1 and 14. Milk levels were comparable to adult female adipose tissue. Data indicate significant transplacental passage and exposure via maternal milk. In rats fed with MX in adulthood, levels were highest in adipose tissue with significant amounts in other organs (ovary, adrenal). Female tissue levels were 3.7-6.8 times higher. This unexplained sex difference was unrelated to lipid content and was absent in offspring. PMID- 9745703 TI - PCDD and PCDF contamination in catfish feed from Arkansas, USA. AB - One combined catfish feed sample from Arkansas, USA, and its eight ingredients were analyzed for PCDDs and PCDFs. One of the ingredients, soybean meal, was highly contaminated by PCDDs, especially the toxic 2,3,7,8-substituted congeners, e.g., 7.3 pg/g dry weight or 370 pg/g lipid for the 2,3,7,8-tetra CDD. The I-TEQ value for the soybean meal was 11.4 pg/g dry weight or 576 pg/g fat. The corresponding values for the combined catfish feed concentrations were approximately 3 times lower. The congener pattern, the congener profile and the ratio sigma PCDDs/sigma PCDFs for the soybean meal were quite unique. We are not aware of any environmental sample or technical product with similar characteristics. As a result, natural formation of the PCDDs found in the soybean meal cannot be ruled out. PMID- 9745706 TI - Isolation of a MX-guanosine adduct formed at physiological conditions. AB - 3-Chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone (MX), a highly potent mutagen present in chlorine-disinfected drinking water, was allowed to react with adenosine, guanosine, and cytidine in aqueous solutions. HPLC analyses, with detection at 254 and 310 nm, showed that a clearly detectable base adduct was formed in the reaction with guanosine. This substance was isolated by C18 column chromatography and characterized by UV absorbance, 1H NMR spectroscopy, and mass spectrometry. The compound was identified as 10-formyl-1, N2-benzoquinone propenoguanosine (I), and the yield was estimated to be approximately 0.1% in reactions performed at pH 7.4 and 37 degrees C. PMID- 9745707 TI - Levels of chlordane compounds in fish muscle, -meal, -oil and -feed. AB - Cis-, trans-, oxychlordane and trans-nonachlor were determined in the edible part of more than 140 fish samples of 15 different species and in fish meal, -oil and fish feed. The investigated fish included most of the important fish species consumed in Germany. Highest concentrations were found in muscle of marine fish with high or moderate fat content, but also in eel and farmed salmon. Marine fish with low fat content contained only traces of chlordane in the muscle tissue. A relationship between fishing ground and levels of chlordane could not be established. Contamination level of herring was related to the age (length) of the fish. Data are also given for contamination levels of fish meal, -oil and feed. PMID- 9745708 TI - The distribution of murine 115-kDa epithelial microtubule-associated protein (E MAP-115) during embryogenesis and in adult organs suggests a role in epithelial polarization and differentiation. AB - In interphase cells microtubules play fundamental roles in the intracellular distribution and movement of organelles and vesicles and thereby contribute to cellular polarization and differentiation. The organization of microtubules varies with the cell type and is presumably controlled by tissue-specific microtubule-associated proteins (MAPs). The 115-kDa epithelial MAP (E-MAP-115) has been identified as a microtubule-stabilizing protein predominantly expressed in cell lines of epithelial origin. To assess a putative function of E-MAP-115 in epithelial morphogenesis in vivo, we have cloned the cDNA encoding the murine protein and studied the cellular distribution of E-MAP-115 mRNA and protein during murine embryogenesis and in adult organs. Analysis of the predicted amino acid sequence of murine E-MAP-115 revealed 81% sequence identity with its human homolog, the best-conserved part of the protein being the microtubule-binding site. Our data indicate that E-MAP-115 is expressed in several epithelia from 9.5 days of embryogenesis onwards and that its expression levels increase during development. From 14.5 days onwards, E-MAP-115 mRNA is found in some neuronal cells as well. In adult organs, E-MAP-115 is most abundant in epithelial cells of kidney tubules, in absorptive cells of the intestine and is widely distributed in the testis. E-MAP-115 expression correlates with the differentiation of certain epithelial cell types: in the adult intestine, for example, E-MAP-115 mRNA and protein are more abundant in the differentiating than in the proliferative cell compartment. Moreover, E-MAP-115 expression clearly correlates with the degree of cellular apicobasal polarity. In the developing kidney, E-MAP-115 mRNA is detected in the cuboidal cells of S-shaped bodies, of primitive tubules and glomerula, whereas, E-MAP-115 mRNA and protein are absent from mature podocytes which have lost their initial apico-basal polarity. The pattern of distribution of E-MAP-115 in vivo is so far unique for a MAP. Taken together, our results provide support for a role of E-MAP-115 in reorganizing the microtubule cytoskeleton during epithelial cell polarization and differentiation. PMID- 9745709 TI - Modulation of laminin integrin receptors in the postnatal and adult rat lung. AB - Recent studies have shown that type II pneumocytes, at birth and day 3 postnatally, have a diffuse distribution and localize at alveolar 'corners' between 3 and 7 days. Since alpha 3 beta 1 and alpha 6 beta 1 are laminin-binding receptors that are well expressed by rat type II alveolar epithelial cells, we postulated that they may play a role in the localization of the cells in the alveolus. To begin the evaluation of this hypothesis, we studied the temporal and spatial expression of the alpha 3, alpha 6, and beta 1 integrin subunit protein and mRNA in whole rat lungs during postnatal development by immunofluorescence, confocal microscopy, and Northern blot analysis. The temporal expression of proteins analyzed by immunochemistry, with integrin subunit specific antibodies, increased during the 3- to 7-day postnatal period and in adult lungs. Densitometric values of the alpha 3, alpha 6, and beta 1 mRNA expression, normalized to 28S rRNA, quadrupled from day 1 to day 3 postnatally. The mRNA expression of different integrin chains was elevated 1.5- to threefold from days 5 to 7 postnatally compared to day 1 levels. The alpha 3 and alpha 6 integrin subunit mRNA decreased to newborn levels in adult lungs, whereas the beta 1 integrin mRNA in adult lungs was expressed at approximately 50% of its level in newborn lungs. We postulate that the increases in alpha 3, alpha 6, and beta 1 integrin mRNA expression during the early neonatal period may be important for the spatial distribution of type II pneumocytes. PMID- 9745710 TI - Adipose tissue extracellular matrix: newly organized by adipocytes during differentiation. AB - The distribution of eight types of extracellular matrix (ECM) proteins (type I VI) collagen, laminin and fibronectin) in the skeletal muscle of Japanese Black cattle was determined by indirect immunofluorescence using specific antibodies against each protein. ECM proteins were well organized in the intramuscular connective tissue: type I, II, III collagen and fibronectin were localized primarily in the perimysium, type V and VI collagen in both the perimysium and endomysium, and type IV collagen and laminin were virtually confined to the endomysium. In the loose connective tissue holding the adipocytes together to form a tissue mass between the muscular bundles, seven of the ECM proteins not type II collagen were relatively abundant in a disordered arrangement. Further analysis by in vitro immunocytochemical staining also demonstrated that a stromal vascular preadipocyte cell line (BIP cell), derived from Japanese Black cattle, synthesized various ECMs in much the same way as fibroblasts. Exponentially growing BIP cells with a fibroblastic phenotype were found to produce type II, V, and VI collagens, in addition to the other previously identified connective tissue glycoproteins of mouse 3T3 preadipocytes. When confluent preadipocyte cultures were stimulated with adipogenic medium, a fibrillar network of ECM was observed to bridge the intercellular space and connect adjacent cell surfaces. During adipocyte differentiation, type III collagen and laminin were arranged in a non-fibrous structure, and type-II collagen was only barely detected. These results are supported by the staining of the adipose tissue, where all ECM proteins studied except type II collagen were stained intensely. These data indicate that in vivo under conditions permissive for adipose conversion, the production and organization of ECM, accompanied by hyperplasia and hypertrophy of precursor cells, gives rise to adipose tissue in skeletal muscle with its own ECM products. These data further suggest that each ECM protein might have some role for the adipocytes in forming tissue. PMID- 9745711 TI - Phenotypic and functional characterization in vitro of a multipotent epithelial cell present in the normal adult human breast. AB - The developmental relationships between the different mammary epithelial cell lineages in the human mammary gland are not well defined. To characterize human breast epithelial cells (HBEC) with progenitor activity, we used flow cytometry and single cell sorting to analyze the distribution of cellular phenotypes in primary cultures of reduction mammoplasties and their associated ability to generate colonies in 2-dimensional (D) and 3-D (collagen gel) culture systems. This approach allowed two distinct types of HBEC progenitor populations to be distinguished on the basis of their differential expression of the MUC-1 glycoprotein, CALLA/CD10 and epithelial-specific antigen (ESA). The first type of progenitor, which is enriched in the MUC-1+/CAL-LA-/ESA+ subpopulation, generated colonies of tightly arranged cells in 2-D cultures and small alveolar-like colonies with a central lumen when cultured in a collagen matrix. The cells produced in the colonies and derived from these MUC-1+/CALLA-/ESA+ progenitors were found to express typical luminal epitopes (keratin 8/18, keratin 19, MUC-1, ESA) and showed low levels of expression of myoepithelial epitopes (keratin 14 and CD44v6). The second type of progenitor, which is present in the MUC-1-to +/ /CALLA +/- to +/ESA+ subpopulation, generated mixed colonies of both luminal and myoepithelial cells when seeded in 2-D and 3-D cultures. In 2-D cultures, the centrally located cells exhibited a luminal morphology and expressed ESA, but were heterogeneous in their expression of MUC-1. Radiating from the periphery of these ESA+ HBEC were highly refractile ESA- teardrop-shaped myoepithelial-like cells. When cultured in a collagen matrix, these bipotent progenitors generated large branched colonies composed of a heterogeneous population of cells, with some of the progeny cells expressing luminal epitopes (keratin 8/18, keratin 19 and MUC-1) and others expressing myoepithelial epitopes (keratin 14 and CD44v6). A third type of progenitor, which became apparent is passaged HBEC cultures and was enriched in the MUC-1-/CALLA+/ESA- subpopulation, was found to generate colonies of cells with an exclusively myoepithelial phenotype. These results provide definitive evidence for the existence of multilineage HBEC progenitors in normal adult human mammary tissue. The phenotypic profile of these cells suggest that these multilineage progenitors are a relatively undifferentiated cell since they express low levels of MUC-1 and that they have a luminal location within the mammary epithelium since they are ESA+. Furthermore, we suggest that the MUC 1+/CALLA-/ESA+ and the MUC-1- to +/-/CALLA +/- to +/ESA+ progenitors we have identified and characterized are candidate in vivo alveolar and ductal progenitors, respectively. PMID- 9745712 TI - Phenotypic characterization of rat hepatoma cell lines and lineage-specific regulation of gene expression by differentiation agents. AB - Hepatoma cell lines can be characterized by their expression of hepatocyte- and biliary-specific genes and by their response to differentiating agents in a lineage-dependent manner. These characteristics can be used to map the maturational lineage position of the cell lines. Tissue-specific gene expression and regulation by heparin, dimethylsulfoxide (DMSO), and sodium butyrate (SB) were examined in three rat hepatoma cell lines and two rat liver epithelial cell lines. Based on antigenic profiles and gene expression in serum-supplemented medium, the hepatoma cell lines could be organized in distinct categories of hepatic differentiation. All three hepatomas expressed the following five genes: gamma-glutamyl transpeptidase (GGT), glutathione-S-transferase pi (Yp), glutamine synthetase, and alpha 5 and beta 1 integrin. Cell line H4AzC2 also expressed alpha-fetoprotein (AFP), albumin. IGF II receptor, and the biliary/oval cell antigens OC.2 and OC.3, a phenotype characteristic of fetal hepatocytes. FTO-2B cells lacked AFP, OC.2, and OC.3 but expressed albumin and IGF II receptor in addition to the five commonly expressed genes, consistent with a more hepatocyte like phenotype. Cell line H5D-7 expressed neither albumin nor the IGF II receptor, but did express OC.2, OC.3, and alpha 3 integrin in addition to the five commonly expressed genes, characteristic of biliary epithelial cells. Regulation of gene expression by heparin, DMSO, and SB was examined in cells cultured in hormonally defined medium. The patterns of regulation of AFP, albumin, GGT, and Yp were dependent upon the state of differentiation of the cell. FTO-2B cells regulated genes in a manner similar to that of E16 fetal hepatocytes, H4AzC2 regulated genes characteristic of both hepatocytic and biliary lineages, and H5D.7 regulated only biliary genes. Suppression of GGT by DMSO was uniformly observed. The three cell lines expressed equal amounts of HNF 4, but FTO-2B cells expressed more HNF-3 beta and less HNF-3 alpha, while the reverse was true of H4AzC2 and H5D.7 cells. PMID- 9745713 TI - Phenotype modulation in primary cultures of aortic smooth muscle cells from streptozotocin-diabetic rats. AB - Diabetes mellitus is a major risk factor for atherosclerosis. In atherosclerotic lesions, arterial smooth muscle cells (SMC) change from a contractile to a synthetic phenotype characterized by active proliferation. A similar phenotype modulation occurs in vitro when isolated arterial SMC are grown in culture and is characterized by both changes in cell morphology and a typical switch in actin isoform expression. In this study, we examined the influence of streptozotocin (STZ)-induced diabetes on the differentiation state and the phenotype modulation of cultured rat aortic SMC. We used transmission electron microscopy to study the fine structure of STZ-diabetic and non-diabetic SMC in primary culture and immunological methods for the determination of the proportions of alpha-smooth muscle actin (alpha-SM) and nonmuscle beta-actin (beta-NM) isoforms. Cultured STZ diabetic SMC exhibited a large cytoplasmic volume, rich in rough endoplasmic reticulum, when compared with cultured non-diabetic SMC. alpha-SM, organized in stress fibers, was less homogeneously and abundantly distributed and by contrast, beta-NM was more abundant in STZ-diabetic than in non-diabetic SMC. Cytofluorimetric analyses demonstrated that the alpha-SM content was reduced in freshly STZ-diabetic SMC. Furthermore, during logarithmic growth of cultured SMC, the decrease of alpha-SM was more important in STZ-diabetic than in non-diabetic SMC. Immunoblotting of actin isoforms confirmed that expression of beta-NM was more important in STZ-diabetic than in non-diabetic SMC even in freshly isolated cells. The results suggest that SMC from STZ-diabetic rats express a more dedifferentiated state and undergo a more rapid phenotypic modulation in primary cultures than SMC from non-diabetic rats. Therefore, diabetes could induce changes in the phenotype of arterial SMC which might be associated with the onset or progression of the atherogenic process. PMID- 9745714 TI - Route of vaccine administration: effects on the specific humoral response in rainbow trout Oncorhynchus mykiss. AB - The specific humoral response of teleost fish to extracellular bacteria was examined using a rainbow trout-Vibrio anguillarum model. Treatment groups were immunized by oral, immersion, and injection routes. All 3 delivery methods conferred full protection in controlled laboratory challenges (p < 0.01). Prior to boosting, serum antibody titers did not correlate with protection in the orally and immersion-vaccinated groups, but, contrary to previous studies, titers measured 10 and 17 d after boosting correlated positively with protection in all 3 vaccinated groups. The route of administration strongly affected the magnitude of the antibody response as measured by enzyme-linked immunosorbent assay (ELISA) and Western blots; however, the antigenic epitopes recognized were not substantially altered by delivery method as evidenced in immunoblot patterns. Given that the primary and booster vaccination protocols were identical, the data suggest that all 3 vaccinated groups may have had a specific humoral response following initial immunization but that specific serum antibody levels before boosting were too low to be detected by ELISA in fish vaccinated by oral and immersion routes. An anamnestic response was evident in all 3 groups. The data support the possibility that teleosts, like higher vertebrates, have a protective immune response to extracellular bacteria that is predominantly humoral. Route of delivery may primarily affect the efficiency with which the immunogenic constituents of the vaccine are presented to the relevant recognition and effector components of the immune system. PMID- 9745715 TI - Oral pharmacological treatments for parasitic diseases of rainbow trout Oncorhynchus mykiss. II. Gyrodactylus sp. AB - A total of 24 drugs were evaluated as regards their efficacy for oral treatment of gyrodactylosis in rainbow trout Oncorhynchus mykiss. In preliminary trials, all drugs were supplied to infected fish at 40 g per kg of feed for 10 d. Twenty two of the drugs tested (aminosidine, amprolium, benznidazole, bithionol, chloroquine, diethylcarbamazine, flubendazole, levamisole, mebendazole, metronidazole, niclosamide, nitroxynil, oxibendazole, parbendazole, piperazine, praziquantel, ronidazole, secnidazole, tetramisole, thiophanate, toltrazuril and trichlorfon) were ineffective. Triclabendazole and nitroscanate completely eliminated the infection. Triclabendazole was effective only at the screening dosage (40 g per kg of feed for 10 d), while nitroscanate was effective at dosages as low as 0.6 g per kg of feed for 1 d. PMID- 9745716 TI - Oral pharmacological treatments for parasitic diseases of rainbow trout oncorhynchus mykiss. III. Ichthyobodo necator. AB - A total of 32 drugs were evaluated as regards their efficacy for oral treatment of Ichthyobodo necator infestation of rainbow trout. In preliminary trials, all drugs were supplied to infected fish at 40 g per kg of feed for 10 d. The majority of the drugs tested (1,3-di-6-quinolylurea, aminosidine, amprolium, benznidazole, bithionol, chloroquine, diethylcarbamazine, dimetridazole, diminazene aceturate, febantel, flubendazole, ketoconazole, levamisole, mebendazole, netobimin, niclosamide, niridazole, nitroscanate, nitroxynil, oxibendazole, parbendazole, piperazine, praziquantel, ronidazole, sulphaquinoxaline, tetramisole, thiophanate, toltrazuril and trichlorfon) were ineffectdive. Metronidazole and secnidazole were 100% effective (unlike the other nitroimidazoles tested, namely dimetridazole, benznidazole and ronidazole). The non-carbamate benzimidazole triclabendazole was likewise 100% effective. PMID- 9745717 TI - Digenean parasites of the bivalve mollusc Pisidium amnicum in a small river in eastern Finland. AB - The host-parasite relationship between digeneans and a semelparous population of the mollusc. Pisidium amnicum Muller in a small river in eastern Finland was studied during 1992/1993. The parasite prevalence of the population was high. The total prevalence was 45.6% in 1992 (n = 790) and 47.5% in 1993 (n = 160). The dominant digenean, Bunodera luciopercae (34.2% in 1992, 35.0% in 1993), had highest prevalences in July/August and in winter. Two other species, Palaeorchis crassus (7.8% in 1992, 7.5% in 1993) and Phyllodistomum elongatum (4.7% and 5.0%), were rare during the winter. The prevalence of B. luciopercae increased as clams aged, while the other species were most common in middle-sized clams. Apparently B. luciopercae rediae dominate over P. elongatum, which has only sporocyst stages, while P. crassus, which has large rediae, is more deleterious to the clam and induces host mortality. Double infections were significantly less common (1.2%) than might be expected by chance. All parasites castrated their hosts; no clam containing both parasites and embryos was found. Semelparity of the population is apparently caused by parasitic castration. PMID- 9745719 TI - Characterization of a non-occluded baculovirus-like agent pathogenic to penaeid shrimp. AB - A non-occluded baculovirus-like agent recently isolated by this laboratory from moribund Penaeus japonicus shrimps obtained from China and named Chinese baculovirus (CBV) was purified and some of its properties characterized. Under the electron microscope, negatively stained virus particles were rod-shaped, enveloped, and measured 322 to 378 nm in length and 130 to 159 nm in diameter. The nucleoprotein core exhibited a unique striated structure and measured 316 to 350 nm in length and 65 to 66 nm in diameter. The striations appear to be the result of the stacking of ring-like structures. These rings consisted of 2 rows of 12 to 14 globular subunits. Each globular subunit measured approximately 10 nm in diameter. SDS-PAGE gels of purified virus preparations showed, among several, 4 prominent protein bands with approximate molecular weights of 19, 23.5, 27.5 and 75 kDa. The structural viral proteins were identified by western blot analysis using polyclonal hyperimmune serum made against purified CBV. The 19, 27.5, and 75 kDa structural proteins were determined to be non-glycosylated components associated with the viral envelope. The 23.5 kDa protein, also non glycosylated, was identified with the capsid structure. Viral genomic DNA digested with Hind III restriction endonuclease revealed at least 29 different fragments with a conservatively estimated total size of at least 183 kb. PMID- 9745718 TI - Prevalence and distribution of QPX, Quahog Parasite Unknown, in hard clams Mercenaria mercenaria in Virginia, USA. AB - In July 1996, the Virginia Institute of Marine Science initiated a sampling program to examine wild and cultured hard clams Mercenaria mercenaria for QPX, Quahog Parasite Unknown, a protistan parasite associated with severe mortalities of hard clams in localized areas in maritime Canada and Massachusetts, USA. The sampling program set out to seasonally monitor wild clams from one site, James River, Virginia, and cultured clams from 2 sites, Chincoteague Bay and Mattawoman Creek, Virginia. Histological examination of initial samples revealed 8% prevalence of the parasite in 1-2 yr old cultured clams in Chincoteague Bay. This is the first documentation of QPX in Virginia. To ascertain the distribution of the parasite in Virginia, the survey was expanded between August 1996 and July 1997 to include 16 additional sites. A total of 1305 wild and cultured clams was sampled from Chesapeake Bay tributaries and coastal areas where harvest and culture occur. QPX was not found in Chesapeake Bay, but was present in cultured clams from 3 coastal embayments--the original Chincoteague Bay site, Burton Bay and Quinby Inlet. The parasite was found in Chincoteague Bay at each sample period at prevalences ranging from 8 to 48%. Infections were generally light to moderate intensity and were most often observed in mantle and gill tissues. The maximum prevalence was observed in May 1997 and coincided with notable clam mortalities. QPX prevalences at the other sites were low, ranging from 4 to 15%. To date QPX has not had a significant impact on Virginia's hard clam fishery and aquaculture industry; however, the presence of the pathogen in 3 of the state's most productive hard clam growout areas warrants continued monitoring and research. PMID- 9745720 TI - A comparative study of three different isolates of white spot virus. AB - Three separate isolates of white spot virus (WSV) purified from 3 different penaeid shrimp species from different countries were compared morphologically, biochemically, and genomically using the following techniques; negative stain electron microscopy, sodium dodecyl-sulfate polyacrylamide gel electrophoresis/western blot, and restriction fragment length polymorphism (RFLP), respectively. Under the electron microscope, the 3 isolates were indistinguishable. Their nucleoprotein cores exhibited the unique striated structure characteristic of the baculovirus-like agents associated with white spot syndrome. The dimensions of the nucleoprotein cores were also identical for all 3 isolates. SDS-PAGE gels of purified virus preparations showed all 3 to be identical in the position of at least 3 of the most prominent protein bands of WSV, with approximate molecular weights of 19, 23.5, and 27.5 kDa. Western blot analyses also revealed these 3 same protein bands in identical positions for all 3 isolates. RFLP analyses of the viral genomes using Hind III and EcoR 1 enzymes revealed that although the 3 isolates were identical when cut with EcoR I, the isolate from Penaeus japonicus from China was distinguishable from the other 2 genomes (P. monodon from Indonesia and P. setiferus from the U.S.) when cut with Hind III. PMID- 9745721 TI - Hyperplastic growth of mucous cells in the mantle of the mussel Modiolus kurilensis from a heavily polluted area of Amursky Bay, Sea of Japan. AB - This histology of an external mantle growth in 1 of 500 Modiolus kurilensis from a heavily polluted area of Amursky Bay, Sea of Japan, was described. The growth consisted of subepithelial basophilic mucous cells containing glucosaminoglycans, eosinophilic large-granular cells with proteins and neutral polysaccharides, and mixed (acid + neutral mucopolysaccharides) cell types. Some subepithelial eosinophilic and basophilic gland cells were dividing and seemed to be the source of tumor growth. The mitotic index of growth cells reached 0.5% on some growth sections; however, many mitoses were pycnotic. The emergence of the tumor on the mussel mantle is probably related to a compensatory or regenerative hyperplasia of subepithelial mucous cells. PMID- 9745722 TI - Rapid molecular mass and structural determination of plant cell wall-derived oligosaccharides using off-line high-performance anion-exchange chromatography/mass spectrometry. AB - A method has been developed for the rapid molecular mass determination and structural elucidation of mixtures of oligosaccharides derived from plant cell walls. The oligosaccharides were fractionated using gel permeation chromatography and 'analytical' high-performance anion-exchange chromatography (HPAEC), neutralized, dried and the mixtures of eluent salt and oligosaccharides were per O-acetylated directly. The derivatized oligosaccharides were isolated by dissolution in dichloromethane and the salts were removed by aqueous partitioning. The per-O-acetylated oligosaccharides were analysed using electrospray (ES) and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MS). Exploiting the fact that acid-catalysed per-O-acetylation of oligosaccharides can be achieved even under the extremely salty conditions that are found in post-column neutralized HPAEC fractions, and combining this derivatization step with off-line ESMS, allow rapid screening for molecular mass and thus yield information on the composition of the various oligosaccharides in these complex mixtures. Subsequent per-O-methylation of the per-O-acetylated, salt-free fractions and collision-induced dissociation tandem mass spectrometric analysis was used for additional sequence and branching determination of the oligosaccharides. PMID- 9745723 TI - Rapid identification and speciation of Haemophilus bacteria by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. AB - Several species of the genus Haemophilus are well known etiological agents of pneumonia, meningitis, conjunctivitis, epiglottitis and chancroid. However, identification and speciation of Haemophilus is both time consuming and labor intensive. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI/TOF-MS) has been used by several investigators to profile proteins from intact and disrupted bacteria; consequently, MALDI/TOF-MS has emerged as a powerful tool in diagnostic bacteriology. This paper reports the use of MALDI/TOF-MS as a technique for the rapid identification and speciation of Haemophilus. This technique was used to not only identify the pathogen, H. ducreyi, but also to determine strain differences from different isolates. Mass spectral 'fingerprints' were obtained which permitted the rapid speciation of not only pathogenic forms of Haemophilus, but also those bacteria which are normally regarded as non-pathogenic and members of the normal flora. MALDI/TOF mass spectra can be acquired in 10 min, allowing the identification of Haemophilus spp. within 24 h rather than the 48 h or more needed for traditional bacteriological methods. In addition, these are the first mass spectral fingerprints available in the literature for many of these organisms. PMID- 9745724 TI - Calcium regulation of phototransduction in vertebrate rod outer segments. AB - The biochemical events underlying the phototransduction cascade in retinal photoreceptors of vertebrates are now well established, on the basis of a wealth of electrophysiological and biochemical evidence. In this review the Ca2+ regulation of the enzymes that generates the photoreceptor light response is analyzed, as well as the Ca2+ transport across the plasma membrane. Most of the results discussed in the following were collected from electrophysiological experiments. PMID- 9745725 TI - A novel photosensitizer, 2-butylamino-2-demethoxy-hypocrellin A (2-BA-2-DMHA). 1. Synthesis of 2-BA-2-DMHA and its phototoxicity to MGC803 cells. AB - The reaction of hypocrellin A (HA) with n-butylamine in pyridine under reflux leads to the formation of 2-butylamino-2-demethoxy-hypocrellin A (2-BA-2-DMHA), which is illustrated by ultraviolet-visible absorption spectra, proton nuclear magnetic resonance spectra, infrared spectra and mass spectra. The product exhibits stronger red-light absorption and has a much higher photopotentiation factor than HA (i.e., more than 200 versus four at a dose of 4 J cm-2 of red light on human gastric adenocarcinoma MGC803 cells). The mechanism of phototoxicity of 2-BA-2-DMHA on MGC803 cells irradiated with red light (lamada = 600-700 nm) has also been studied. An examination of extracted cellular DNA by agarose gel electrophoresis shows that the DNA has degraded into fragments with lengths which are multiples of approximately 180-190 base pairs (i.e., oligonucleosome size), a biochemical marker of apoptosis. Transmission electron microscopy reveals chromatin condensation around the periphery of the nucleus, which is also characteristic of apoptosis. This study suggests that 2-BA-2-DMHA is a potential photosensitizer and that its photoxicity to MGC803 cells proceeds via apoptosis. PMID- 9745726 TI - 5-Aminolaevulinic acid-induced protoporphyrin IX accumulation in tissues: pharmacokinetics after oral or intravenous administration. AB - In this study, the biodistribution of 5-aminolaevulinic acid (ALA) and accumulation of protoporphyrin IX (PpIX) in rats have been examined. Two groups of 21 WAG/Rij rats are given 200 mg/kg ALA orally or intravenously. Six rats serve as controls. At 1, 2, 3, 4, 6, 12 and 24 h after ALA administration, ALA and porphyrin concentrations are measured in 18 tissues and fluids. Liver enzymes and renal-function tests are measured to determine ALA toxicity. In both groups ALA concentration is highest in kidney, bladder and urine. After oral administration, high concentrations are also found in duodenal aspirate and jejunum. Mild, short-lasting elevation of creatinine is seen in both treatment groups. Porphyrins, especially PpIX, accumulate mainly in duodenal aspirate, jejunum, liver and kidney (> 10 nmol/g tissue), less in oesophagus, stomach, colon, spleen, bladder, heart, lung and nerve (2-10 nmol/g tissue), and only slightly in plasma, muscle, fat, skin and brain (< 2 nmol/g tissue). In situ synthesis of porphyrins rather than enterohepatic circulation contributes to the PpIX accumulation. Confocal laser scanning microscopy shows selective porphyrin fluorescence in epithelial layers. Peak levels and total production of porphyrins are equal after oral and intravenous ALA administration. IN CONCLUSION: administration of 200 mg/kg ALA results in accumulation of photosensitive concentrations of PpIX, 1 to 6 h after ALA administration, in all tissues except muscle, fat, skin and brain. Knowledge of the time-concentration relationship should be helpful in selecting dosages, routes of administration and timing of ALA photodynamic therapy. PMID- 9745727 TI - Characterization of psoralen-oleic acid cycloadducts and their possible involvement in membrane photodamage. AB - By UVA irradiation of an ethanol solution of psoralen and oleic acid, four main photoproducts have been isolated and characterized: two have cis,cis structure; the other two are trans,cis. The same adducts have been isolated from the photoreaction of psoralen with beta-oleoyl-gamma-stearoyl-1-alpha phosphatidylcholine followed by enzymatic hydrolysis with phospholipase A2. The four isomers stimulate protein kinase C to almost the same extent. PMID- 9745728 TI - Phenanthroline-Cu complex-mediated chemiluminescence of DNA and its potential use in antioxidation evaluation. AB - The chemiluminescence (CL) concomitant with phen-Cu2+/ascorbate/H2O2-induced DNA damage has been studied. The emission intensity increases linearly with increasing DNA concentration. The emission spectrum has a maximal wavelength at about 410 nm. The luminescence is inhibited by histone in a histone concentration dependent manner. The CL is characteristic of guanine. Of all common kinds of bases and nucleotides, only guanine or guanine nucleotides can give rise to luminescence. The possibility of using the luminescence as a means of studying antioxidation related to DNA damage is discussed. Several kinds of well-defined antioxidants have been used as testing reagents and show that this method can not only evaluate the antioxidative effect but also distinguish different types of antioxidants. PMID- 9745729 TI - Mexoryl SX: a broad absorption UVA filter protects human skin from the effects of repeated suberythemal doses of UVA. AB - There is now considerable evidence that chronic UVA exposure induces damage in animal and human skin; however, little is known about UVA protection of human skin. The aim of this study is to evaluate the efficacy of Mexoryl SX, a broad UVA absorber (lamada max = 345 nm) against UVA-induced changes in human skin. The regimen of UVA exposure (13 weeks with increasing suberythemal doses) induces intense pigmentation with no erythema. Skin hydration and elasticity decrease, whereas total skin thickness, assessed by echography, remains unchanged. Irradiated epidermis reveals a significant thickening of the stratum corneum, an absence of hyperplasia and an increase in the expression of the protective iron storage protein ferritin. No significant alterations are seen using antisera against type IV collagen or laminin, suggesting that the dermal-epidermal junction (DEJ) is mainly preserved. In dermis, enhanced expression of tenascin is seen just below the DEJ but type I procollagen, which is localized at the same site, is unaltered. Although we are unable to visualize any changes in elastic network organization using Luna staining or specific antiserum directed against human elastin, we notice an increased deposition of lysozyme or alpha-1 antitrypsin on elastin fibres. Mexoryl SX (5%) efficiently prevents these alterations. Thus, these results suggest that UVA photoprotection can prevent early putative alterations leading to photoageing. PMID- 9745731 TI - Phantom echo generation: a new technique for investigating dolphin echolocation. AB - In behavioral experiments where real targets are used to investigate dolphin echolocation, it is often very difficult to extract the relevant echo parameters that the animals use to discriminate or classify. The complex relationship between the physical dimensions and the reflection characteristic of real targets prevents separate control of various echo parameters of the stimuli presented in an echolocation experiment. A new echo simulation method presented in this paper avoids this problem. Dolphin echolocation sounds are transformed with the target impulse response into artificial echoes, which are played back to the animal. The phantom echo system is implemented on a digital signal processing board and gives an experimenter fully programmable control over the echo generating process and the echo structure itself. Echoes of several underwater targets were simulated to evaluate the quality of the method. A comparison of simulated echoes with the original echoes demonstrated very good agreement independent of the incident signal (cross-correlation coefficient > 0.95). The method has tremendous potential for investigating animal echolocation and understanding biosonar signal processing. PMID- 9745730 TI - Herpes simplex virus proteins are damaged following photodynamic inactivation with phthalocyanines. AB - The photodynamic inactivation of herpes simplex virus type 1 (HSV-1) by two phthalocyanines (Pcs), the cationic dye HOSi-PcOSi(CH3)2(CH2)3N+(CH3)3I-(Pc5) and the amphiphilic dye aluminum dibenzodisulfophthalocyanine hydroxide (AlN2SB2POH), has been compared with that by the anionic dye, Merocyanine 540 (Mc540). Both Pc derivatives demonstrate a remarkable virucidal activity upon light activation even 3 h after the onset of HSV-1 adsorption, while Mc540 is effective for only 30 min after adsorption. Since fusion and virus penetration are promoted by membrane glycoproteins, we have studied the damage to viral proteins following photodynamic treatment (PDT) of HSV-1 and its relation to inactivation. The effect of AlN2SB2POH PDT is assessed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Major changes are found in the protein profile of PDT treated HSV-1. A reduced ability of specific antibodies to react with HSV-1 major envelope proteins is detected by employing the Western blot assay. In particular, we demonstrate the related changes of glycoprotein D (gD), a structural protein of the HSV envelope. Since the envelope proteins participate in viral entry into the host cell, these alterations to viral envelope proteins may impair their ability to participate in early events of viral entry, leading to reduced infectivity of HSV-1. In contrast, no significant changes in the proteins' electrophoretic mobility could be seen after PDT with Mc540 or with Pc5. When HSV 1 purified proteins are subjected to combined electrophoretic and electro osmotic forces on cellulose acetate, there is a shift in their cathode mobility, which may indicate changes in the protein mass and protein net charges following AlN2SB2POH photosensitization. There are only minor changes in the virus proteins, assayed as above, when HSV-1 is treated with Pc5. PMID- 9745732 TI - Flow generated around particle clusters in a rotating ultrasonic waveguide. AB - A chamber cavity, which has a square cross section and pressure-release walls, is used to produce a well-defined, 160-kHz standing ultrasonic field. A suspension of latex microspheres in aqueous metrizamide fills the chamber. The chamber rotates about a horizontal axis producing the centripetal force necessary to contain the buoyant spheres in the axial region. At low particle concentrations, clusters of microspheres form at half-wavelength intervals near the axial positions of acoustic pressure amplitude (p0) minima, as expected because of rotational and acoustic radiation forces. At higher concentrations, additional particle distributions are often seen that suggest the presence of flow. When high concentrations of larger particles are used, small clusters also form at axial positions of p0 maxima. Theory for acoustic streaming in a rotating fluid predicts flow speeds that are too small to account for the observed flow. Reasonable agreement with observations is obtained using a theory for flow generated by the buoyant gravitational force acting on the clusters. PMID- 9745733 TI - Model experiment to study sonic boom propagation through turbulence. Part II. Effect of turbulence intensity and propagation distance through turbulence. AB - A model experiment was reported to be successful in simulating the propagation of sonic booms through a turbulent atmosphere [B. Lipkens and D. T. Blackstock, J. Acoust. Soc. Am. 103, 148-158 (1998)]. In this study the effect on N wave characteristics of turbulence intensity and propagation distance through turbulence are investigated. The main parameters of interest are the rise time and the peak pressure. The effect of turbulence intensity and propagation distance is to flatten the rise time and peak pressure distributions. Rise time and peak pressure distributions always have positive skewness after propagation through turbulence. Average rise time grows with turbulence intensity and propagation distance. The scattering of rise time data is one-sided, i.e., rise times are almost always increased by turbulence. Average peak pressure decreases slowly with turbulence intensity and propagation distance. For the reported data a threefold increase in average rise time is observed and a maximum decrease of about 20% in average peak pressure. Rise times more than ten times that of the no turbulence value are observed. At most, the maximum peak pressure doubles after propagation through turbulence, and the minimum peak pressure values are about one-half the no-turbulence values. Rounded waveforms are always more common than peaked waveforms. PMID- 9745734 TI - Lamb wave assessment of fiber volume fraction in composites. AB - Among the various techniques available, ultrasonic Lamb waves offer a convenient method of examining composite materials. Since the Lamb wave velocity depends on the elastic properties of a material, an effective tool exists to evaluate composites by measuring the velocity of these waves. Lamb waves can propagate over long distances and are sensitive to the desired in-plane elastic properties of the material. This paper discusses a study in which Lamb waves were used to examine fiber volume fraction variations of approximately 0.40-0.70 in composites. The Lamb wave measurements were compared to fiber volume fractions obtained from acid digestion tests. Additionally, a model to predict the fiber volume fraction from Lamb wave velocity values was evaluated. PMID- 9745735 TI - Linear and nonlinear model of the human middle ear. AB - The measurement of the middle ear transfer function usually requires invasive methods. An equivalent analog equivalent model enables us to evaluate its characteristics without damaging any part of the ear. A linear and a nonlinear model of the middle ear have been developed to predict intracochlear pressure and the stapes volume velocity for various sound pressure levels (SPL). The linear model results have been compared with human eardrum impedance and middle ear transfer function data. The nonlinear phenomena due to the contraction of the stapedius muscle over 80 dB and to the stapes clipping displacement above 120 dB are represented by a set of variable electrical components. The model of the acoustic reflex is based on experimental observations. The study of the annular ligament behavior was performed on cats and extrapolated to humans with some hypothetical restrictions. These approximations provide information on the middle ear transfer function and enable us to better understand the nonlinear middle ear mechanisms in an intense acoustic field. PMID- 9745736 TI - Modeling otoacoustic emission and hearing threshold fine structures. AB - A class of cochlear models which account for much of the characteristic variation with frequency of human otoacoustic emissions and hearing threshold microstructure is presented. The models are based upon wave reflections via distributed spatial cochlear inhomogeneities and tall and broad cochlear activity patterns, as suggested by Zweig and Shera [J. Acoust. Soc. Am. 98, 2018-2047 (1995)]. They successfully describe in particular the following features: (1) the characteristic quasiperiodic frequency variations (fine structures) of the hearing threshold, synchronous and click-evoked emissions, distortion-product emissions, and spontaneous emissions; (2) the relationships between these fine structures; and (3) the distortion product emission filter shape. All of the characteristic frequency spacings are approximately the same (0.4 bark) and are mainly determined by the phase behavior of the apical reflection function. The frequency spacings for spontaneous emissions and threshold microstructure are predicted to be the same, but some deviations from these values are predicted for synchronous and click-evoked and distortion-product emissions. The analysis of models is aided considerably by the use of the solutions of apical, and basal, moving solutions (basis functions) of the cochlear wave equation in the absence of inhomogeneities. PMID- 9745737 TI - Enhancement of electrically evoked oto-acoustic emissions associated with low frequency stimulus bias of the basilar membrane towards scala vestibuli. AB - Electrically evoked oto-acoustic emissions (EEOAEs) are sounds present in the ear canal when ac current is passed into the cochlea. EEOAEs are attributed to the activation of fast electromotile responses in outer hair cells (OHCs). An interesting property of EEOAEs is the phenomenon of "acoustic enhancement," where the emission amplitude is increased by moderate-level sound [D. C. Mountain and A. E. Hubbard, Hear. Res. 42, 195-202 (1989)]. In this report a form of enhancement is described which occurs with displacements of the basilar membrane toward scala vestibuli, during amplitude modulation of the EEOAE waveform by low frequency tones. This "SV-bias enhancement" possibly consists of two components: (i) a low-level component induced by sound at levels which produce nonlinear growth of the cochlear microphonic and which may be equivalent to the "acoustic enhancement" described previously, and (ii) a high-level component which occurs at sound levels well above those which cause saturation of the cochlear microphonic. The low-level component could be explained by either an increased access of the extrinsically applied current to a membrane-based source of OHC motility, perhaps coupled with a reduction in negative feedback, or an increase in electromotile output during scala vestibuli displacements, but the origin of the high-level component is obscure. PMID- 9745738 TI - Changes in otoacoustic emissions in a transsexual male during treatment with estrogen. AB - Otoacoustic emissions (OAEs) were monitored in two human males undergoing estrogen treatment prior to sex-reversal surgery. In one subject, multiple spontaneous emissions (SOAEs) appeared where none had been evident previously. One reasonable interpretation is that (in this male, at least) androgens normally produced a suppressive effect on the cochlear mechanisms responsible for SOAEs, and that the decline in androgen levels produced by the estrogenic drug led to a reduction in that suppression. PMID- 9745739 TI - The auditory evoked potential difference tone and cubic difference tone measured from the inferior colliculus of the chinchilla. AB - The auditory evoked potential f2-f1 difference tone (DT) and the 2 f1-f2 cubic difference tone (CDT) were recorded from electrodes implanted in the inferior colliculus in a group of chinchillas. The purpose of this study was to measure normative aspects of AEP distortion products in awake chinchillas, by comparing the DT and CDT under a variety of stimulus conditions. For experiment 1, f1 was held constant at 1998 Hz, while the f2/f1 ratio was varied from 1.05 to 1.50. Input-output functions were measured over a range of primary tone levels up to 80 dB SPL. The amplitude of the DT was greatest for the smallest f2/f1 ratio, and decreased systematically as f2/f1 ratio increased. DT amplitude was greater than CDT amplitude for all primary tone pairs. Experiment 2 was conducted to determine the effect of f1 frequency upon the DT and CDT for a constant f1-f2 difference frequency of 102 Hz (f1-999, 1998, 4999, and 9998 Hz). The DT input-output functions were overlapping for all f1 frequencies. For the CDT, amplitude decreased with increasing f1 frequency, which corresponded to an increase in CDT frequency. In experiment 3, the relationship between ear of stimulation and inferior colliculus recorded from was investigated. DT input-output functions (f1 = 1998 Hz, DT = 102 Hz) were measured for monaural contralateral, monaural ipsilateral, and dichotic stimulus conditions. DT amplitude was largest for the contralateral condition, followed by the ipsilateral condition. A smaller, dichotic component to the DT was observed as well. PMID- 9745740 TI - Lateralization of large interaural delays. AB - Two experiments explored the limits of listeners' abilities to interpret large interaural time delays (ITDs) in terms of laterality. In experiment 1, just noticeable differences (jnd's) were measured, using an adaptive procedure, for various reference ITDs of Gaussian noise between 0 and 3000 microseconds. The jnd's increased gradually with reference ITD for reference ITDs between 0 microsecond and 700 microseconds, then rose sharply to plateau at much higher jnd's for the remainder of the standard ITDs tested (1000-3000 microseconds). The second experiment tested left/right discrimination of Gaussian noise that was interaurally delayed up to 10,000 microseconds, and high-pass filtered to cutoff frequencies between 0 Hz (broadband) and 3000 Hz. There was good discrimination (62%; significantly above chance, p < 0.05) for broadband and 500-Hz high-pass cutoff stimuli for all ITDs up to 10,000 microseconds, and for ITDs up to at least 3000 microseconds for higher high-pass cutoff frequencies. These results indicate that laterality cues are discriminable at much larger ITDs than are experienced in free-field listening, even in the absence of energy below 3 kHz. PMID- 9745741 TI - Frequency-weighting functions for broadband speech as estimated by a correlational method. AB - The relative contributions of various regions of the frequency spectrum to speech recognition were assessed with a correlational method [K. A. Doherty and C. W. Turner, J. Acoust. Soc. Am. 100, 3769-3773 (1996)]. The speech materials employed were the 258-item set of the Nonsense Syllable Test. The speech was filtered into four frequency bands and a random level of noise was added to each band on each trial. A point biserial correlation was computed between the signal-to-noise ratio in each band on the trials and the listener's responses, and these correlations were then taken as estimates of the relative weights for each frequency band. When the four bands were presented separately, the correlations for each band were approximately equal; however, when the four bands were presented in combination, the correlations were quite different from one another, implying that in the broadband case listeners relied much more on some bands than on others. It is hypothesized that these differences reflect the way in which listeners combine and attend to speech information across various frequency regions. The frequency-weighting functions as determined by this method were highly similar across all subjects, suggesting that normal-hearing listeners use similar frequency-weighting strategies in recognizing speech. PMID- 9745742 TI - Auditory-visual spatial integration: a new psychophysical approach using laser pointing to acoustic targets. AB - The alignment of auditory and visual spatial perception was investigated in four experiments, employing a method of laser pointing toward acoustic targets in combination with various tasks of visual fixation in six subjects. Subjects had to fixate either a target LED or a laser spot projected on a screen in a dark, anechoic room and, while doing so, direct the laser beam toward the perceived azimuthal position of the sound stimulus (bandpass-filtered noise; bandwidth 1-3 kHz; 70 dB sound pressure level, duration 10 s). The sound was produced by one of nine loudspeakers, located behind the acoustically transparent screen between 22 degrees to the left and 22 degrees to the right of straight ahead. Systematic divergences between sound azimuth and laser adjustment were found, depending on the instructions given to the subjects. The eccentricity of acoustic targets was generally overestimated by up to 10.4 degrees with an only slight influence of gaze direction on this effect. When the sound source was straight ahead, gaze direction had a substantial influence in that the laser adjustments deviated by up to 5.6 degrees from sound azimuth, toward the side to which the gaze was directed. This effect of eye position decreased with increasing eccentricity of the sound. These results can be explained by the interactive effects of four distinct factors: the lateral overestimation of the auditory eccentricity, the effect of eye position on sound localization, the effect of the retinal eccentricity on visual localization, and the extraretinal effect of eye position on visual localization. PMID- 9745743 TI - Validity of rating scale measures of voice quality. AB - The validity of perceptual measures of vocal quality has been neglected in studies of voice, which focus more commonly on rater reliability. Validity depends in part on reliability, because an unreliable test does not measure what it is intended to measure. However, traditional measures of rating reliability only partially represent interrater agreement, because they cannot reflect variations or patterns of agreement for specific voice samples. In this paper the likelihood that two raters would agree in their ratings of a single voice is examined, for each voice in five previously gathered data sets. Results do not support the continued assumption that traditional rating procedures produce useful indices of listeners' perceptions. Listeners agreed very poorly in the midrange of scales for breathiness and roughness, and mean ratings in the midrange of such scales did not represent the extent to which a voice possesses a quality, but served only to indicate that listeners disagreed. Techniques like analysis by synthesis or judgment of similarity avoid decomposing quality into constituent dimensions, and do not require a listener to compare an external stimulus to an unstable internal representation, thus decreasing the error in measures of quality. Modeling individual differences in perception can increase the variance accounted for in models of quality, further reducing the error in perceptual measures. Thus such techniques may provide valid alternatives to current approaches. PMID- 9745744 TI - Speech intelligibility assessment in a helium environment. II. The speech intelligibility index. AB - The Speech Intelligibility Index (SII) was measured for Navy divers participating in two saturation deep dives and for a group of nondivers to test different communication systems and their components. These SIIs were validated using the Speech Perception in Noise (SPIN) test and the Griffiths version of the Modified Rhyme Test (GMRT). Our goal was to determine if either of these assessments was sensitive enough to provide an objective measure of speech intelligibility when speech was processed through different helmets and helium speech unscramblers (HSUs). Results indicated that SII values and percent intelligibility decreased incrementally as background noise level increased. SIIs were very reliable across the different groups of subjects indicating that the SII was a strong measurement for predicting speech intelligibility to compare linear system components such as helmets. The SII was not useful in measuring intelligibility through nonlinear devices such as HSUs. The speech intelligibility scores on the GMRT and SPIN tests were useful when the system component being compared had a large measurable difference, such as in helmet type. However, when the differences were more subtle, such as differences in HSUs, neither the SPIN nor the GMRT appeared sensitive enough to make such distinctions. These results have theoretical as well as practical value for measuring the quality and intelligibility of helium speech enhancement systems. PMID- 9745745 TI - Artificial buzzing lips and brass instruments: experimental results. AB - Experimental results of a special artificial trombone player are presented: A mechanical device is a substitute for the musician. Wind instruments, and particularly the brass, are self-sustained oscillators. The oscillations are induced by a mechanical oscillator (the lips of the player) acting as a valve which modulates the flow. Measured mechanical parameters of the artificial buzzing lips for different "embouchures of the player" are presented, and analyzed in connection with the played frequencies obtained for the same "embouchures." The results are obtained with two resonator systems (a mouthpiece alone and a trombone with its mouthpiece). PMID- 9745746 TI - Simple methods of estimating source levels and locations of marine animal sounds. AB - This paper describes three relatively simple methods of estimating source levels of marine animal sounds by estimating the source distance acoustically, using one or two hydrophones. The methods are logistically simpler than using arrays of hydrophones of known positions but are accurate over a smaller range of distances. One method makes use of the differences in the arrival times and levels of the signals received at two hydrophones from the same sound emission. No knowledge of the positions of the hydrophones is required, however, if these are known the position of the source can be determined, with left-right ambiguity. The second method uses the difference in received levels only, but requires the hydrophone spacing to be known. Adequate accuracy requires the source to be significantly closer to one hydrophone than to the other. Third, if the direct and surface reflected arrivals can be separated, the source level can be determined with a single hydrophone. The methods require accurately calibrated hydrophones. PMID- 9745747 TI - Characterizing the graded structure of false killer whale (Pseudorca crassidens) vocalizations. AB - The vocalizations from two, captive false killer whales (Pseudorca crassidens) were analyzed. The structure of the vocalizations was best modeled as lying along a continuum with trains of discrete, exponentially damped sinusoidal pulses at one end and continuous sinusoidal signals at the other end. Pulse trains were graded as a function of the interval between pulses where the minimum interval between pulses could be zero milliseconds. The transition from a pulse train with no inter-pulse interval to a whistle could be modeled by gradations in the degree of damping. There were many examples of vocalizations that were gradually modulated from pulse trains to whistles. There were also vocalizations that showed rapid shifts in signal type--for example, switching immediately from a whistle to a pulse train. These data have implications when considering both the possible function(s) of the vocalizations and the potential sound production mechanism(s). A short-time duty cycle measure was developed to characterize the graded structure of the vocalizations. A random sample of 500 vocalizations was characterized by combining the duty cycle measure with peak frequency measurements. The analysis method proved to be an effective metric for describing the graded structure of false killer whale vocalizations. PMID- 9745748 TI - The effect of spatial separation of signal and noise on masking in the free field as a function of signal frequency and age in the mouse. AB - Masking of low- (4 kHz) and high-frequency (25 kHz) signals by one-octave bandpass maskers either spatially coincident with the signal or contralateral to it was examined in mice, 4-6 and 20-22 months of age, in the free field. Signals were presented 120 ms prior to a startle stimulus and differences in their inhibition of the startle reflex, relative to startle stimulus alone trials, were used to measure the severity of masking. Inhibition was reduced or eliminated by spatially coincident noise for weak but not for relatively intense signals, providing the type of "loudness recruitment" effect characteristic of human listeners in similar stimulus conditions. The spatial separation of the signal and its masker relieved this maskinglike effect for the high-frequency pair in both young and old mice. In contrast there was no beneficial effect of the shift in spatial location for the low-frequency pair at either age. This finding of masking release for high- but not low-frequency stimuli supports the hypothesis that the sound shadow provided by the head and pinna would yield a favorable signal-to-noise level difference for a contralateral masker and an ipsilateral signal only at very high frequencies in the mouse. The presence of masking release in these old mice, a first generation hybrid strain with near-normal high frequency hearing in ABR measures, agrees with reports that the masking release resulting from a similar manipulation in aged human listeners with minimal high frequency hearing loss is the equal of that obtained in the young listener. PMID- 9745749 TI - Changes in temporal acuity with age and with hearing impairment in the mouse: a study of the acoustic startle reflex and its inhibition by brief decrements in noise level. AB - Temporal acuity for brief gaps in noise was studied in mice of different ages (1 36 months) from strains with differing susceptibility to age-related hearing loss, using reflex modification audiometry. Prepulse inhibition of the acoustic startle reflex (ASR) increased with gap depth (GD: 10-40 dB in 70 dB SPL noise) and lead time (LT: 1-15 ms). The increase in inhibition with LT followed an exponential function in which the two parameters, asymptotic inhibition (AINH) and the time constant (tau), were both affected by GD. AINH rapidly declined from 1 to 6 and then to 18 months of age in C57BL/6J mice with progressively severe hearing loss, but first increased with maturation and then gradually declined beyond 6-12 months of age in CBA/CaJ and CBA x C57BL Fl-hybrid mice, which show no apparent change in sensory function at these ages. In contrast, tau was unaffected by hearing loss or by age, this suggesting that age-related changes in this form of temporal acuity occur because of a reduction in the efficiency with which gaps are centrally processed, not from any reduced ability to follow their rapid shift in noise level. PMID- 9745750 TI - Focusing of therapeutic ultrasound through a human skull: a numerical study. AB - A numerical model was developed which can use digitized layer interfaces to calculate ultrasound wave absorption, diffraction, reflection, and refraction. This model was used to evaluate the feasibility of ultrasound therapy and surgery through a human skull. A digitized human skull profile was obtained from magnetic resonance (MR) images and used to calculate the ultrasound field in the brain of a volunteer from a spherically curved phased array. With no phase correction, the focus of the array was shifted and defocused. The phased array technique was used to correct focal shift, reduce side lobes, and enhance focal amplitude. The optimum source element width was estimated for each frequency to obtain a near optimium focus, and an appropriate frequency range for transskull ultrasound therapy and surgery was determined. Acoustic pressure amplitude on the skull surfaces was examined, and it was shown that the skull heating problem could be overcome. Despite high attenuation, complex interface shape, and nonuniform thickness of a human skull, a sharply focused transskull ultrasound field can be generated for noninvasive ultrasound therapy and surgery in the brain. PMID- 9745751 TI - Effects of high-frequency amplification on double-vowel identification in listeners with hearing loss. PMID- 9745753 TI - Lyme disease: self-regulation and pathogen invasion. AB - Ecological interactions underlying the epidemic of Lyme disease involve a spirochete, a tick (with larval, nymph and adult stages), and two (or more) vertebrate hosts. Juvenile ticks ordinarily feed on mice; adult ticks feed on deer. Mice acquire the spirochete from infected nymphs and then pass the infection to larvae of the next tick generation. Lyme disease may result when a human is inadvertently bitten by an infectious nymph. Our model of the Lyme phenomenon counts the total number of ticks in each stage, the numbers of infected ticks by stage, and the number of infected mice. We fix the total population sizes of deer and mice, assume the ticks self-regulate, and solve the homogeneous-mixing case for equilibrium abundances. A local stability analysis identifies a condition where extinction of the spirochete is stable. Reversing this condition implies that the spirochete can invade the system of ticks and vertebrate hosts. When the spirochete can invade, a positive equilibrium number of infected organisms is locally stable. Spirochete invasion is promoted by a sufficient density of mice suffering low mortality, high susceptibility to infection in both mice and ticks, a high attack rate of ticks on mice, a high density of larval ticks, and low mortality among tick nymphs. Low mouse mortality allows the frequency of infection among nymphs to approach an individual tick's susceptibility when feeding on an infected mouse. PMID- 9745752 TI - A mathematical model for ligand/receptor/G-protein dynamics and actin polymerization in human neutrophils. AB - A mathematical model is proposed for describing the dynamics of the chemotactic peptide-stimulated actin polymerization response in human neutrophils. The response pathway utilizes the guanine nucleotide binding protein (G-protein) signal transduction cascade common to many receptor systems and allows adaptation in the continued presence of ligand. The development of such a model is an important first step toward understanding, predicting, and ultimately manipulating neutrophil responses. The model is divided into two parts, ligand/receptor/G-protein dynamics and the actin polymerization mechanism. Fast (receptor precoupled to G-protein) and slow (free receptor) signaling pathways involving ligand/receptor/G-protein interactions produce an activated signaling molecule. The actin polymerization mechanisms utilizes an actin binding protein which complexes with actin monomer and inhibits polymerization in an unstimulated cell. During stimulation, the activated signaling molecule enhances the dissociation of monomer/binding protein complexes, allowing the actin polymerization response to occur. The fast and slow signaling pathways are predicted to have different roles in controlling the time course of this actin polymerization. Additionally, precoupled receptors are predicted to have a larger ligand association rate constant than non-precoupled (free) receptors. Model simulations agree with many of the experimentally observed characteristics of both the stimulated F-actin response and ligand/receptor binding kinetics for both the fluorescent peptide ligand CHO-norleucyl-leucyl-phenylalanyl-norleucyl tyrosyl-lysine-fluorescein (CHO-NLFNTK-fl) and the non-fluorescent peptide ligand CHO-methionyl-leucyl-phenylalanine (CHO-MLF). PMID- 9745754 TI - Networks with side branching in biology. AB - There are many examples of branching networks in biology. Examples include the structure of plants and trees as well as cardiovascular and bronchial systems. In many cases these networks are self-similar and exhibit fractal scaling. In this paper we introduce the Tokunaga taxonomy for the side branching of networks and his parameterization of self-similar side-branching. We introduce several examples of deterministic branching networks and show that constructions with the same fractal dimension can have different side-branching parameters. As an example of stochastic-branching in biology we consider the vein structure of a leaf. We show that the vein structure of the leaf and river networks have nearly identical side branching statistics. We introduce diffusion limited aggregation (DLA) clusters. These clusters also exhibit Tokunaga side-branching statistics. We consider several alternative explanations for why leaves, river networks, and DLA clusters have similar side-branching statistics. We also consider the allometric scaling relation between metabolic rate and the mass of species in terms of a cardiovascular system with Tokunaga statistics. We find reasonably good agreement between the observed scaling exponent, approximately 0.75, and our model for a range of values of the fractal dimension of the network and the blood flow resistance parameter. PMID- 9745755 TI - Non-equilibrium thermodynamics of molecular evolution. AB - The evolution of the information complexity of a large database of protein sequences is investigated. The information entropy for protein sequences is determined from their algorithmic complexity and is found to change with evolutionary time at a constant rate. The information content of changed residues is always lower than the content of conserved residues. This indicates that sequences are becoming less random throughout evolution. It also shows that the system is being driven toward minimal complexity production. The change in information content per amino acid substitution is virtually identical for all the protein sequences studied. These results are interpreted with a statistical mechanical theory that ties sequence information to the thermodynamics of protein structure. Sequence evolution is viewed as a means to drive the system to minimum thermodynamic entropy production in a stable, non-equilibrium state. This theory provides a physical framework for understanding molecular evolution and incorporates features of both the neutralist and selectionist models. PMID- 9745757 TI - Optical feedback controlled scleral remodeling as a mechanism for myopic eye growth. AB - Experimental studies of myopia have demonstrated that optical errors imposed on a developing eye will stimulate elongation of the globe, although the mechanism of axial growth is not well understood. In this study, a mathematical model of eye growth is presented in which expansion of the globe occurs as the elastic deformations of the scleral shell are incorporated into the zero stress configuration of the eye during the scleral remodeling process. The rate of remodeling is determined by the retinal blur and the amplitude of accommodation, which provide feedback loops for both unilateral and bilateral hyperopic refractive error-compensation. Normal eye growth and experimental myopia are simulated in tree shrews, a small mammal related to primates. The model demonstrates that the rate of ocular elongation in experimental myopia may be controlled by regulating the rate of soft tissue remodeling in the scleral shell. PMID- 9745756 TI - Immune avoidance strategies in RNA viruses: fitness continuums arising from trade offs between immunogenicity and antigenic variability. AB - Highly exposed and protruding amino acid sites on the surface of viral capsids are subject to fewer residue interactions and packing constraints than those buried within protein interiors. Consequently they often experience higher rates of non-synonymous substitution and exhibit greater genetic variability than buried interior sites. However such protrusive surface structures often induce host immune responses and are likely to constitute B cell epitopes. Genetic variation at these surface sites is therefore likely to correspond to antigenic variation. This may be of adaptive value to the virus for two quite different reasons. The first is that antigenic variation arising over the course of a viraemia may result in greater net viral replication, and increased opportunities for viral transmission. The second is that antigenic variation generated rapidly over a single infection or incrementally over several sequential infections may give rise to variants that are sufficiently immunologically distinct that they can reinfect host individuals with previous infection experience of related virus. This would lead to an extension of the susceptible host pool with consequent increase in transmission opportunities. The surface architecture of viral capsid proteins is therefore conceivably subject to two opposing selection pressures: one to minimize the surface area accessible to interaction with elements of the immune system, the other to increase the potential access to antigenic variation by adoption of exposed and unconstrained protein conformations. Therefore, there exists a possible trade-off between the fitness benefits deriving from potential ability to generate antigenic variation, and the increased immunogenicity with which such potential may be associated. We propose that the existence of this trade-off would lead to a continuum of different strategies by which a virus might combat an immune response. We explore this strategy space with simple mathematical models, and show that peak loads of infectious virus particles are proportional to levels of antigenic diversity, and inversely proportional to immunogenicity, thereby creating the potential for a trade-off by which fitness might be maintained with a continuum of strategies. This may remain possible even if the antigenic variants are not transmissible between hosts, so long as immune resources are sufficiently dispersed between antigenic variants. The diversity of possible strategies is discussed with reference to the Picornavirus family. PMID- 9745758 TI - Micromechanical models for the Brownian motion of hair cell stereocilia. AB - Brownian motion of the hairs (stereocilia) of amphibian hair cells has been shown in experiments to be in the range of some nm. Our models of the Brownian motion of coupled harmonic oscillators with mechanical properties of stereocilia lead to similar displacements. Computer simulation shows that stochastic fluctuations enhance the encoding of low level acoustic signals. Stochastic resonance lowers the detection threshold of auditory signals to amplitudes one order of magnitude lower than that of the Brownian motion. PMID- 9745759 TI - Control of cell proliferation by progress in differentiation: clues to mechanisms of aging, cancer causation and therapy. AB - In multicellular organisms, the control of cell proliferation occurs, in part, by modulating the progress in differentiation. In normal and neoplastic cells, for example, progress towards terminal differentiation concludes cell proliferation, whereas arrest of progress in differentiation causes uncontrolled cell proliferation. Evidence is presented according to which the progress in differentiation depends on an increase in the ratio between mitochondrial differentiation promoting activity and nuclear differentiation preventing activity. This ratio is low in embryonic cells and in stem cells, due to low mitochondrial content, but increases by a rate of mitochondrial multiplication that is larger than a doubling of mitochondrial content per cell cycle. The rate of mitochondrial multiplication, thus, decides on the progress in differentiation and controls the number of amplification divisions between cell determination and terminal differentiation. This rate is modifiable by extracellular signals and cellular defects. Mutations, for example in nuclear genes coding for mitochondrial proteins, are likely to decrease the rate so much that differentiation is arrested with ensuing neoplastic growth. Agents used in differentiation therapy and ionizing radiation overcome this arrest: the cell cycle is sensitive to these agents, but not the mitochondria which multiply during the transitory cell cycle inhibition, thus increasing the differentiation promoting activity. Differentiation arrest can be circumvented also by direct inhibition of nuclear differentiation preventing activity at the level of transcription or translation, whereas corresponding inhibition of mitochondrial differentiation promoting activity prevents differentiation. Accumulation of non specific genetic damage causes persisting cell cycle prolongation and enhancement of differentiation which, apparently, are involved in senescence. The recent finding of increase in mitochondrial mass prior to release of cytochrome c, induction of differentiation, and apoptosis points to similarities in the initial molecular pathways of differentiation and apoptosis. PMID- 9745760 TI - Amino acid contribution to the genetic code structure: end-atom chemical rules of doublet composition. AB - Up-to-date knowledge is not yet enough to prove or reject completely one of the leading alternative hypotheses considering external or internal origin of the genetic code organization (term "internal" means a peculiarity of amino acids and nucleotides which can propose some logical basis for explanation why definite codons are assigned to particular and not to other amino acids; term "external" in this context implies tRNAs and aminoacyl-tRNA synthetases which provide accuracy of translation but are not presented in the code table in the evident from). New data in favour of the internal approach have been proposed. A close correlation exists between chemical nature of single, the most remote from C alpha, end atoms of amino acid side chains and types of bases in appropriate codon doublets. There are two main rules reflecting this correspondence: (1) all O- or N-ended amino acids (i.e. with O, OO, N, NN, ON, OC and NC end patterns) possess the A-containing codon doublets; (2) all solely C-ended (excluding Ala) and S-ended amino acids (i.e. residues with C, CC and S ends) have the U containing codon doublets. The discovered asymmetry is characterized by the following high reliable differences of A and U distribution among 21 patterns of individual doublets belonging to 18 amino acids (Pro and Gly are discarded): (a) proportion of A plus U bases corresponding to the two rules, 22/26 is much more than proportion of such bases in violating cases, 4/26; (b) A and U are dominant bases over each of other base types in the doublets of O/N- and C/S-ending amino acids, respectively; (c) the numbers of doublets bearing (18) and not bearing (3) dominant A or U bases are also significantly different. The concept of indicative amino acid end atoms (O/N and non-O/non-N) and doublet A/U bases has been proposed and discussed as so far unknown regulative system of submolecular level of heredity which, additionally to possible key role in the process of the genetic code arising, provides now control of its universality, stability and fidelity. PMID- 9745762 TI - Biological effects of high ultraviolet radiation on early earth--a theoretical evaluation. AB - The surface of early Earth was exposed to both UVC radiation (< 280 nm) and higher doses of UVB (280-315 nm) compared with the surface of present day Earth. The degree to which this radiation environment acted as a selection pressure on organisms and biological systems has rarely been theoretically examined with respect to the biologically effective irradiances that ancient organisms would receive. Here action spectra for DNA inactivation and isolated chloroplast inhibition are used to estimate biologically effective irradiances on archean Earth. Comparisons are made with present day Earth. The theoretical estimations on the UV radiation screening required to protect DNA on archean Earth compare well with field and laboratory observations on protection strategies found in present day microbial communities. They suggest that many physical and biological methods may have been effective and would have allowed for the radiation of life even under the high UV radiation regimes of archean Earth. Such strategies would also have provided effective reduction of photoinhibition by UV radiation. The data also suggest that the UV regime on the surface of Mars is not a life limiting factor per se, although other environmental factors such as desiccation and low temperatures may contribute towards the apparent lack of a surface biota. PMID- 9745761 TI - Do microbes with peptides mimicking myelin cause multiple sclerosis if the T cell response to their unique peptides is limited? AB - This hypothesis for the pathogenesis of multiple sclerosis is based upon assumptions about the response of the T cell repertoire to pathogens. Immunologic and epidemiologic observations of several conditions suggest that activation of T cells formed in early life mediate injury to the central nervous system. Early in life, selection of lymphocytes by the thymus produces a weakly autoreactive T cell repertoire which, with the help of transient maternally-derived defenses, recognizes pathogens. These responses later are supplemented by pathogen-specific responses, acquired as microbes are encountered. As the thymus involutes, the diversity of pathogen-specific responses to microbial epitopes is progressively fixed. Reduced and delayed pathogen exposure, common in developed societies, limits the repertoire of memory T cells, which can efficiently eliminate pathogens. Due to their small number, pathogen-specific lymphocytes which mature extrathymically may not be able to rapidly eliminate most pathogens, and without the editing of the thymus, they may be autoreactive. In this setting, novel pathogens with epitopes mimicking myelin may elicit a T cell response which is autoreactive. Peptides of common microbes are known to activate T cells recognizing dominant antigens of myelin. It is postulated that at the equator, intense, non-seasonal encounters with microbes elicit an immune repertoire that produces resistance to autoimmunity, while, in temperate climates, moderate, seasonal exposures increase susceptibility to it. The differences in responses to microbes between populations with a low or high prevalence of multiple sclerosis suggests that T cell repertoires are divergent in these groups. An exuberant innate response, postulated to diminish as the load of enteric microbes falls and sanitation improves in relation to the distance from the equator, may increase resistance to multiple sclerosis by eliminating the need for T cell activation. Human herpesvirus-6 and respiratory syncytial virus are possible prototypes of microbes which activate myelin-directed T cells. PMID- 9745763 TI - Scaling properties of the placenta's arterial tree. AB - The purpose of the present work is to establish a basic knowledge about the scaling properties of the placenta's arterial tree. For this end we have analysed X-ray angiograms of 22 normal arterial trees by box counting. All the investigated arterial trees scale closely according to a power-law over a one decade wide range of scales. Perfectly self-similar fractals, of the same resolution as our representation of the arterial tree, do not follow a power-law more closely. The results support the hypothesis that a mechanism or rule--as regular as those which dictate the structure of perfectly self-similar fractals- also determines fundamental aspects of the arterial tree's morphology. PMID- 9745764 TI - Evolution of angiosperms via modulation of antagonisms. AB - The controversies concerning the evolutionary mechanisms of flowering plants continue unabated in spite of the current trends toward the analysis of macromolecular data and of the growing body of distributional micromolecular data. The usual narrative approach to this latter effort is here replaced by a novel technique, quantitative phytochemistry. An evolutionary outline emerges and reveals modulation of antagonisms as the fundamental mechanism of angiosperm evolutionary ecology. Origin or operation of many systems can be rationalized analogously. It is concluded that the impact of opposing features possesses universal relevance. PMID- 9745765 TI - Iridoids from Loasa acerifolia. AB - In the course of preliminary chemotaxonomic studies on Loasaceae from South and Central America, 30 species from 6 genera were screened by chromatographic means. From the aerial parts of Loasa acerifolia Dombey, the novel trimeric and dimeric iridoid glucosides acerifolioside and tricoloroside methyl ester, consisting of loganin and secoxyloganin moieties, were isolated. The structures were primarily elucidated by NMR spectroscopy. Loganin, loganic acid, rutin, scopoletin and chlorogenic acid were identified by cochromatography with reference compounds. PMID- 9745767 TI - Lignans and sesquiterpene lactones from Artemisia sieversiana and Inula racemosa. AB - The aerial parts of Artemisia sieversiana afforded, in addition to beta sitosterol, stigmasterol and daucosterol, two novel lignans as well as one known and three new guaianolides. The roots of Inula racemosa gave beta-sitosterol, daucosterol and isoalantolactone. The structures were determined by a combination of spectral methods (IR, EIMS, 1H and 13C NMR, DEPT, COSY, NOESY and HETCOR). All isolates were subjected to antifungal tests. Isoalantolactone, a major sesquiterpene lactone of I. racemosa, was found to be active against the human pathogenic fungi. Aspergillus flavus, A. niger, Geotrichum candidum, Candida tropicalis and C. albicans at concentrations of 50, 50, 25, 25 and 25 micrograms/ml, respectively. The taxonomic significance of the characterized constituents is discussed briefly. PMID- 9745766 TI - Enzymatic hydrolysis of the cytotoxic triterpenoid glycoside virgaureasaponin 1. AB - The cytotoxic compound, virgaureasaponin 1, was converted using several optimized enzymecatalysed hydrolyses to the 28-O-beta-D-xylopyranosyl (1-->4)-alpha-L rhamnopyranosyl-(1-->2)-beta-D-fucopyranoside (2), and the 28-O-alpha-L rhamnopyranosyl-(1-->3)-beta-D-xylopyranosyl-(1-->4)-alpha- L- rhamnopyranosyl-(1 ->2)-beta-D-fucopyranoside (3) and 28-O-beta-D-xylyopyranosyl-(1-->4)-alpha-L rhamnopyranosyl-(1-->2) - beta-D-fucopyranoside (4) both lacking the glucose moiety at C-3 of the aglycone. The terminal rhamnose of the acylglycosidic bonded tetrasaccharide was cleaved by naringinase to give compound 2. The new acylglycosides 3 and 4 were obtained with the help of a relatively crude beta glucuronidase preparation, but the cleavage of the sapogenin bonded glucose was impossible using several beta-glucosidase preparations directly. These derivatives were used for the investigation of the relationship between the saponin carbohydrate structure and their cytotoxic activity. PMID- 9745768 TI - Modern management of childhood diabetes: a role for computerized devices? AB - Technology is increasingly relevant in everyday life and it can be very interesting to apply it also in the field of diabetes as it can be one way of ensuring better care. In the Diabetes Control and Complications Trial, the intensively treated patients were seen fortnightly in hospital and contacted even more frequently by telephone during the day as well as at night. For these reasons, health reforms are undergoing a radical change in an attempt to reduce the spiraling costs of health care provision. Therefore, it can be useful to use information technology (IT) in diabetes care. The European Federation for Medical Informatics recently established some Special Issues to advance IT initiatives that may be able to address the problem of adjusting the insulin dose and controlling blood glucose (BG) levels, as well as assisting in the provision of modern-day diabetes care. The Special Issues can be classified under the following headings: databases, algorithms, decision support, models and education. It can be useful to develop a prototype computer system that can be applied to different areas of clinical diabetes care; computerized out-patients' clinical databases that store patients' clinical and biochemical data; statistical and graphical analysis programs that help the clinician; dietary analysis programs which examine food composition and dietary exchanges and help devise meal plans; hand-held insulin dosage adjustment computers that advise patients on a day-by-day or even meal-by-meal basis; expert systems and question and answer programs for patients' education, and finally games for children. These systems have the potential to be useful tools in many aspects of diabetes care but their utilization by the vast majority of the health care community has been extremely limited. Nonetheless, computers cannot substitute for the pediatric diabetes team, which remains the major determinant for better care of diabetes in children and adolescents. PMID- 9745770 TI - Assessment of D-glucose transport kinetics in the perfused human placenta: an in vitro study. AB - BACKGROUND: There have been no reports to date on glucose transport kinetics and the effect of graded hyperglycemia in the human placenta in non-steady-state conditions. METHODS: The transport kinetics of D-glucose in the human placenta was investigated in non-steady state conditions in vitro using perfusion of isolated placental lobules. National Cancer Tissue Culture (NCTC) 135 culture medium, diluted with Earle's buffered salt solution was used as the perfusate. 14C-Labeled D-glucose and water as reference were injected as a single bolus into the maternal arterial perfusate. Perfusate samples were collected and analyzed from the maternal and fetal venous outflows. RESULTS: In four successful perfusions, differential transport rates of glucose in the maternal-fetal direction averaged 1.03, 1.02, 1.09, 1.04 and 1.03 times those of corresponding tritiated water transport rates for 10, 25, 50, 75 and 90% of efflux fractions, respectively. Glucose transport fraction, expressed as percentage of injected maternal dose averaged 84 +/- 3.1% of water transport fraction in the four perfusions. Glucose kinetic parameters expressed as area under the curve, elimination constant (Kel), clearance, time for maximum response, absorption rate and elimination rate averaged 0.25, 0.29, 3.96, 1.02, 0.25 and 0.18 times that of the corresponding tritiated water value, respectively. Neither the different kinetic parameters nor the transport fraction indices differed significantly when glucose concentrations in the same perfusions were raised successively from 5.56 to 27.80 and then to 55.6 mmol/L. CONCLUSIONS: We speculate that within physiological limits, hyperglycemia per se plays no significant part in altering glucose transport kinetics across the human placenta in the maternal-fetal direction. PMID- 9745769 TI - Role of excitatory aminoacids in neonatal hypoglycemia. AB - BACKGROUND: In many neurological disorders, injury to neurons may be due in part to overstimulation of the receptors for the excitatory amino acids glutamate and aspartate. The same excitotoxic mechanism and high aspartate levels in experimental studies led to this study of the concentrations of glutamate and aspartate and zinc, copper, and magnesium levels in the cerebrospinal fluid (CSF) of hypoglycemic newborns. METHODS: Aspartate and glutamate were determined by high-performance liquid chromatography, and magnesium, zinc and copper by atomic absorption spectrophotometer. RESULTS: The CSF levels of aspartate (3.98 +/- 1.77 mumol/L) and glutamate (1.7 +/- 1.05 mumol/L) in 20 hypoglycemic newborns were significantly higher when compared with the values of aspartate (2.19 +/- 0.6 mumol/L) and glutamate (0.77 +/- 0.34 mumol/L) of 10 control newborns. In the hypoglycemic patients, the concentration of zinc (0.57 +/- 0.13 microgram/mL), but not copper (0.39 +/- 0.40 microgram/mL) was significantly lower when compared with the control values. There was no difference in the magnesium levels between the two groups. CONCLUSIONS: The higher levels of excitatory amino acids found in the CSF of hypoglycemic infants than in controls were consistent with previous animal studies, which may indicate the role of excitatory amino acids in the late biochemical effects of hypoglycemia in newborn brain metabolism. PMID- 9745771 TI - Modification of the MTT method for the study of bilirubin cytotoxicity. AB - BACKGROUND: We propose a modification of the MTT [3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide] method to study the cytotoxicity of bilirubin. The original method involves reading the intensity of a purplish blue color resulting from the conversion of MTT to formazan crystals by the mitochondria of viable cells. We have found that when the method is applied to study the effect of bilirubin on growing cells, precipitation of the yellow bilirubin pigment interferes with the colorimetric reading. METHODS: A human liver cell line was used. The interference of bilirubin deposition on the MTT assay was investigated by comparing the value of optical density of the MTT solution in the presence and absence of bilirubin. The effect of 0.04 mol/L HCL-isopropanol on the bilirubin precipitate was tested by recovering the amount of bilirubin from the wells after the isopropanol treatment. RESULTS: Bilirubin deposition increases MTT reading by 10-24%. Hydrochloride-isopropanol (0.04 mol/L) dissolves MTT formazan only without disturbing the bilirubin precipitates. The bilirubin extracted into the supernatant was less than 5% of the total bilirubin deposited. DISCUSSION: This indirect MTT assay, as developed in this study, could eliminate the interference of bilirubin deposits and serve as a good method for the study of bilirubin cytotoxicity. PMID- 9745772 TI - Retrospective analysis of clonality and detection of residual disease in myeloid leukemia by FISH on long-term stored bone marrow smears. AB - BACKGROUND: Fluorescence in situ hybridization (FISH) has allowed the detection of numerical chromosomal aberrations in interphase nuclei on fresh or frozen smears of leukemia. METHODS: To analyze clonality and residual disease in myeloid leukemia retrospectively, we applied FISH to bone marrow smears stored at ambient temperature for up to 9 years. RESULTS: When hybridization efficiency was investigated on stored control smears from patients without hematological malignancy, more than 96% of nuclei showed the expected number of signals using DNA probes specific for chromosome 7, X or Y. In combination with cell morphology, we observed much higher hybridization efficiency in blasts and granulomonocytic cells compared with lymphoid and erythroid cells. On the basis of good hybridization efficiency for old smear specimens, we applied FISH to stored bone marrow smears of myeloid leukemias, in which either loss of chromosome 7 or loss of sex chromosomes had been verified previously by conventional cytogenetics (one patient with chronic myelomonocytic leukemia (CMML) and four with acute myeloid leukemia (AML; three M2 and one M7)). As a result, the loss of chromosome was detected in blasts from all patients and was observed in mature granulocytes, except in M7. In the CMML patient and one AML (M2) patient with t(8;21), lymphoid and erythroid cells also showed the loss of chromosomes, suggesting that it should occur at stem-cell level. A high amount of residual disease was detected in the morphological remission samples in one AML (M2) patient after induction therapy. The patient eventually succumbed to relapse. CONCLUSION: Thus, the present FISH technique is useful to analyze the clinical significance of clonality and the residual disease in myeloid leukemia, retrospectively. PMID- 9745773 TI - Mutation in the gene encoding the fibroblast growth factor receptor-3 in Korean children with achondroplasia. AB - BACKGROUND: Achondroplasia (ACH) is the most common form of osteochondrodysplasia, and is mostly associated with a point mutation in the gene on the transmembrane domain of fibroblast growth factor receptor-3 (FGFR-3) on chromosome 4p. METHODS: We investigated the mutations in the gene encoding FGFR-3 in 15 Korean children with ACH, using polymerase chain reaction (PCR) coupled with direct sequencing. RESULTS: In this study, all children with ACH showed the same mutation as those reported in France, USA and Japan; a G-->A transition at position 1138 of the coding sequence, resulting in the substitution of arginine for glycine at position 380 of the mature protein. CONCLUSIONS: This consistent point mutation of Korean children with ACH indicates there is no significant racial difference in the pathogenesis of ACH, compared with data from Caucasian and Japanese children with ACH. PMID- 9745774 TI - Serodiagnosis of infectious mononucleosis in children. AB - BACKGROUND: Although anti-viral capsid antigen (VCA)-immunoglobin M (IgM) is the most reliable serological marker of primary Epstein-Barr virus (EBV) infection, it could only be detected in limited cases of infectious mononucleosis in children. We analyzed anti-EBV antibodies by an enzyme-linked immunosorbent assay (ELISA), a sensitive method for detecting IgM antibody and compared these results with those obtained by a conventional indirect immunofluorescence (IF) method. METHODS: Anti-Epstein-Barr virus early antigen (EA)-IgM and nuclear antigen 1 (EBNA1)-IgG were examined by an ELISA assay in 180 sera from 70 infants and children with infectious mononucleosis, diagnosed serologically by standard IF methods. RESULTS: Although by IF, VCA-IgM was detected in only 37 of 70 (52.9%) of the sera from the acute phase of the disease, by ELISA, EA-IgM was detected in 65/70 (92.9%) of these sera. Among infants less than 12 months of age. EA-IgM was positive in 11/13 cases (84.6%) while VCA-IgM was detected in only 3/13 cases (23.1%). Anti-Epstein-Barr virus nuclear antigen 1-IgG was not detected by ELISA in the sera from the acute phase of infectious mononucleosis. Anti-EBNA was not detected by IF in about one-third of the sera during 6-8 months after onset of the disease, whereas by ELISA, EBNA1-IgG was detected in 93.0%. Sera that were positive or negative for both EA-IgM and EBNA1-IgG by ELISA were observed in several cases after the patients recovered from the disease. CONCLUSIONS: Although serodiagnosis by the combination of ELISA for EA-IgM and EBNAI-IgG was more sensitive than IF methods, especially in the case of infants and young children, several patients during convalescence and recovery might be judged as seronegative or as being in highly reactivated states. PMID- 9745775 TI - Serum levels of C3 and factors I and B in minimal change disease. AB - BACKGROUND: Relapses are an important problem in minimal change disease, which accounts for most of the cases of childhood nephrotic syndrome. Because of defects in the humoral immune system, patients are predisposed to infection in nephrotic syndrome and infection is the most important complication that determines mortality and morbidity. METHODS: In this study, serum levels of Factors I and B and C3 were studied to evaluate the relationships between nephrotic syndrome and infection in 17 children with nephrotic syndrome (24-96 months of age) and 10 healthy children (27-84 months of age). RESULTS: Serum levels of Factors I and B were found to be lowered in the active disease group compared with the control group. These values were lowest for the infection group. Although it was observed that these values increased with steroid treatment, they did not reach normal levels. The parameters in remission were not different from the parameters in the control subjects. The serum level of C3 was found to be high during the active disease state and returned to normal levels during remission. CONCLUSIONS: The patients with active minimal change disease had infections such as peritonitis, septicemia and urinary tract infection because of low concentrations of Factors I and B in their sera. PMID- 9745776 TI - Perinatal management of congenital complete atrioventricular block: report of nine cases. AB - BACKGROUND: The addition of fetal ultrasonography has allowed the prenatal diagnosis and observation of congenital complete atrioventricular block (CCAVB). Thus, the management of the affected fetuses should be modified accordingly. METHODS: The medical records were reviewed to identify patients with CCAVB, and clinical and laboratory data were collected. RESULTS: Nine patients with CCAVB, diagnosed prenatally, were identified. The gestational age at the time of diagnosis ranged from 18 to 38 weeks (median, 25 weeks). The gestational age and birthweight ranged from 29 to 41 weeks and from 1420 to 4075 g, respectively. An accompanying congenital cardiac anomaly was noted in three cases, including one with polysplenia syndrome. The heart rate at birth ranged from 30 to 80 bpm. In three neonates, isoproterenol was given for bradycardia. One fetus with hydrops fetalis associated with complex heart disease was treated with maternally administered digoxin, with resolution of the fluid accumulation. A permanent pacemaker was implanted in six cases: two within 1 day of birth; two during the neonatal period; one at 51 days; and the other at 2 years and 11 months of age. There were no deaths or major complications and all have remained well beyond their infancy. CONCLUSIONS: With the improvement of diagnostic methodology, more cases with CCAVB, including those with complex heart disease, could be diagnosed during fetal life. Their outcome has been steadily improving with the advances in perinatal management, pacemakers and implant techniques, irrespective of associated structural cardiac defects. PMID- 9745777 TI - Renal functional reserve in insulin dependent diabetic children. AB - BACKGROUND: Microalbuminuria has been shown to be predictive for clinical diabetic nephropathy. Renal functional reserve (RFR), as a response to protein loading in a short period of time, is a parameter to assess the ability of kidneys to increase the glomerular filtration rate (GFR). The aim of this study was to predict the early phase of diabetic nephropathy by measuring urinary albumin level and RFR capacity in patients with insulin-dependent diabetes mellitus (IDDM). METHODS: Twenty-two patients with IDDM were studied: 11 with a disease duration of less than 5 years (group 1) and 11 with a disease duration of more than 5 years (group 2). As the control group, 15 healthy children (group 3) were included in the study. At the beginning of the study, glucose was measured and the urinary albumin/creatinine ratio was calculated. Average glycosylated hemoglobin (HbA1c) over 1 year was determined. After protein loading (red meat containing 2 g/kg of protein), the creatinine clearance was calculated at each hour for a duration of 4 h. The RFR was accepted as the peak percentage increase in GFR over the baseline value. RESULTS: Although metabolic control in group 2 was better, the RFR in group 2 was significantly lower than in group 1 (P < 0.05). Urinary microalbumin levels between the groups did not differ (P > 0.05). In two patients in whom microalbuminuria was detected, the RFR was much lower. CONCLUSIONS: Detecting lower RFR levels in patients with normal urinary albumin excretion, as well as in patients with microalbuminuria, may support the idea that the RFR capacity is more sensitive than microalbuminuria in assessing the early phase of diabetic nephropathy. PMID- 9745778 TI - The effect of live measles vaccines on serum vitamin A levels in healthy children. AB - OBJECTIVE: Serum retinol levels have been shown to be depressed during measles infection. This study aims to demonstrate whether there is any decrease in serum vitamin A level following immunization with live viral vaccine and its relation with vaccine seroconversion in children with measles. Since many children receive measles vaccine alone or in combination with measles-mumps-rubella vaccine, we studied serum vitamin A levels and antibody levels in healthy, well-nourished children before and after immunization with monovalent and combined live attenuated measles vaccine. METHODS: The first group included 21 healthy children between the ages of 9-11 months who received live measles (Schwarz) vaccine. There were also 21 healthy children (range 14-20 months of age) who received measles-mumps-rubella Trimovax (Pasteur Merieux) vaccine. All children were tested for serum vitamin A levels before vaccination, on days 9-14 and 30-42 following both vaccinations. Measles specific antibody levels were also measured on admission and 30-42 days following vaccinations. RESULTS: In both vaccination groups, mean serum vitamin A levels reduced significantly on days 9-14, but increased slightly on days 30-42 in the measles-mumps-rubella vaccinated group (P < 0.05). The baseline and follow-up levels of mean serum vitamin A did not differ between seroconverted and nonseroconverted cases within the measles vaccinated group. CONCLUSION: Serum vitamin A levels are reduced following vaccination with monovalent and combined live attenuated measles vaccines. PMID- 9745779 TI - An evaluation of infants' growth in the Kingdom of Nepal. AB - BACKGROUND: The His Majesty's Government/Japan International Cooperation Agency Primary Health Care Project began in April 1993 in collaboration with the Saitama Prefectural Government, for the purpose of improving the health status of the people in model districts of the Kingdom of Nepal. Growth monitoring is one of the basic methods that defines the health and nutritional status of children. METHODS: Anthropometric indices were measured in 759 children in the Bhaktapur district. We used the World Health Organization prototype growth chart and national growth standard for Japanese children (1990) to analyze the growth data. RESULTS: We found that the average bodyweight growth curve of children up to 4 months of age followed the 50th percentile reference curve. For children of 5-12 months of age, there was a delay in bodyweight gain and the growth curve reached the 3rd percentile curve. For children more than 1 year old, the growth curve moved below the third percentile curve. Catch-up growth did not occur before the children reached 5 years of age. The main causes of catch-up growth being hampered were chronic undernutrition and inadequate nutritional balance. CONCLUSIONS: As this was the first opportunity to evaluate infant growth in this district, the first important consequence of the results was to analyze the causes of growth faltering and failure-to-thrive in Nepalese children. Even more important, was the need to give appropriate counseling on improving feeding and other health-related practices, and the most important consequence of all was to instruct Nepalese health workers that utilizing the growth charts is an integral part of health care. PMID- 9745780 TI - Successful polymerase chain reaction-based diagnosis of fungal meningitis in a patient with chronic granulomatous disease. AB - Meningitis is not a common complication of chronic granulomatous disease (CGD). Here, we present details of a 3-year-old boy with X-linked CGD, who suffered from fungal meningitis. While 19 samplings using conventional cerebrospinal fluid (CSF) cultures failed to detect any organisms, fungal DNA was identified in the CSF by a new polymerase chain reaction (PCR)-based method. The patient recovered without any sequelae after treatment with a combination of antifungal agents, interferon-gamma and granulocyte infusions. This case report demonstrates that fungal meningitis must be included in the differential diagnosis of infections in CGD patients and that the PCR-based detection of fungal DNA is a powerful tool for diagnosis. PMID- 9745781 TI - Bacterial croup caused by Pasteurella haemolytica. AB - We report a case of bacterial croup caused by Pasteurella haemolytica. A nine month-old girl was admitted to our hospital because of abrupt onset dyspnea and unconsciousness. From clinical and laboratory findings, she was diagnosed as having bacterial croup. Pasteurella haemolytica was recovered from her tracheal secretion on admission. To our knowledge, this is the first report of bacterial croup caused by P. haemolytica. PMID- 9745782 TI - Recurrent hepatosplenomegaly and peripheral blood cytopenia, persistent Epstein Barr virus infection and central nervous system manifestation in a patient with lymphadenopathy and low serum uric acid. AB - We describe a 27-month-old boy who was first admitted to our hospital on 7 January 1995 with nasal bleeding. From 6 months of age he has had lymphadenopathy, low levels of serum uric acid, increased levels of serum lactate dehydrogenase and hyper gamma-globulinemia. From the age of 18 months he has had persistent Epstein-Barr virus (EBV) infection (target cells; B cells), recurrent episodes of thrombocytopenia, anemia and hepatosplenomegaly. Dysmobility of the left leg and arm from a central nervous system complication during a relapse with pancytopenia on March 1995, was also observed. Relapses of thrombocytopenia with increases of platelet-associated immunoglobulin G and hepatosplenomegaly have been observed approximately every 2 months, and two relapses of pancytopenia were accompanied with weak positivity of Coombs test and low level of haptoglobin. These recurrent episodes were improved with prednisolone. However, now in June 1997 we have not been able to diagnose what underlies the above clinical symptoms, except that the patient has a persistent EBV infection. PMID- 9745783 TI - An infant case of bilateral small kidneys with both proximal and distal tubular dysfunction. AB - A male infant with bilateral small kidneys associated with both proximal and distal tubular dysfunction, who showed chronic renal failure soon after birth, is reported. He was also noted to have both proximal and distal type of renal tubular acidosis. The small kidneys were thought to be due to renal hypodysplasia associated with bilateral severe vesicoureteral reflux, by radiological findings. An alkalization therapy with chemoprophylaxis seemed to be of benefit in slowing the progression of renal failure in this case. PMID- 9745784 TI - Diffuse brain damage caused by acute twin-twin transfusion during late pregnancy. AB - BACKGROUND: Monoamniotic twinning is a relatively rare event with increased antenatal and perinatal mortality. We describe a brain damage detected in a surviving monoamniotic twin after intrauterine death of the co-twin at 37 weeks of gestation. RESULTS: Severe entanglement and knotting of the umbilical cords was apparent at the time of delivery and a portion of the cord to the dead twin was narrowed significantly. It was suggested that transfer of blood occurred across placental anastomoses from the survivor to the dead fetus, resulting in transient but severe hypovolemia in the survivor. It is difficult to prevent this type of brain damage because the course of acute twin-twin transfusion is very rapid and the damage has already occurred by the time the death of the twin is diagnosed. CONCLUSIONS: We suggest that elective delivery should be considered in cases of monoamniotic twin pregnancies with additional risk factors. PMID- 9745785 TI - An autopsy case of fetal Gaucher disease. AB - BACKGROUND: A case of fetal form of Gaucher disease in a Japanese fetus is presented. RESULTS: A macerated baby showing hydrops fetalis was dissected at 29 weeks of gestation. The fetus was heavier in the body, liver and spleen than a normal fetus at the same gestation period. It also suffered from pericardial effusion and ascites. The diagnosis of Gaucher disease was made by histological and biochemical findings. In microscopical examinations 'Gaucher cells', which were periodic acid-Shiff (PAS)-positive, alcian blue-positive and CD68-positive, existed in the lungs, liver, spleen, thymus, adrenal glands, bone marrow and brain. In thin layer chromatography, a large quantity of glucocerebroside was seen to have accumulated in the patient's organs. PMID- 9745786 TI - Isolated deficiency of glucocorticoids presenting with cholestasis. AB - BACKGROUND: Isolated deficiency of glucocorticoids is characterized by elevated levels of adrenocorticotropin (ACTH) and normal aldosterone production. It is rare for isolated deficiency of glucocorticoids to be associated with liver involvement. A case of an infant with isolated deficiency of glucocorticoids presenting with cholestasis is presented in this article. A male infant on his 39th postnatal day was referred to our hospital for evaluation of prolonged jaundice and convulsion. He had two episodes of hypoglycemic convulsion on postnatal 8th and 39th day, after which he was admitted to our hospital. RESULTS: Physical examination revealed systemic jaundice, hyperpigmentation of the skin, hepatomegaly and splenomegaly on admission. He had normal male genitalia with 3.5 cm of penis and bilateral scrotal testes. Laboratory values were as follows: glucose 45 mg/dL, total biluribin 18.14 mg/dL, direct biluribin 6.54 mg/dL, aspartate aminotransferase 378 IU/L, alanine aminotransferase 46 IU/L, and alkaline phosphatase (ALP) 1302 IU/L. In abdominal ultrasound and biliary tract scanning, extra- and intrahepatic biliary tracts were shown to be normal. Finally, biopsy of the liver revealed cholestasis. An endocrinological evaluation showed high levels of adrenocorticotropin (ACTH, 1000 pg/mL), low levels of cortisol (1 microgram/dL) and normal aldosterone levels. CONCLUSIONS: The diagnosis of cholestasis secondary to isolated glucocorticoid deficiency was suspected with clinical and laboratory findings. Hydrocortisone treatment (30 mg/m2 per day) was initiated after which hyperpigmentation and jaundice decreased and ACTH and ALP levels reduced to 39 pg/mL and 440 IU/l, respectively. We emphasize that cholestasis in infants may be a component of isolated deficiency of glucocorticoids. PMID- 9745787 TI - Diffuse splenic and visceral hemangiomas complicated by chronic consumption coagulopathy. AB - The case of a 7-year-old girl with a 2 year history of easy bruising associated with thrombocytopenia is reported. On admission she presented with ecchymoses, abdominal distention and splenomegaly. Hemostasis investigation revealed a consumption coagulopathy. Several radiological studies failed to confirm the diagnosis of diffuse splenic and visceral hemangiomatosis, which was eventually established by an explorative laparotomy. Platelet count and the other coagulation abnormalities progressively returned to normal after splenectomy, although the remaining hemangiomas were extensive. PMID- 9745788 TI - Prevalence of GBV-C/HGV infection in pregnant Japanese women. AB - Recently, a novel viral agent, hepatitis G virus, was identified by independent researchers from the serum of patients with liver disease, and termed GBV-C or HGV. At present, GBV-C and HGV are considered to be separate isolates of the same virus; however, the role of this virus in acute and chronic liver disease remains uncertain. Although vertical transmission is known to be one of the routes of transmission, the prevalence of GBV-C/HGV viremia in pregnant Japanese women is unknown. Thus, we determined this prevalence using the reverse transcription polymerase chain reaction (RT-PCR). PMID- 9745789 TI - CT based 3D Monte Carlo radiation therapy treatment planning. AB - This paper outlines the "voxel reconstruction" technique used to model the macroscopic human anatomy of the cranial, abdominal and cervical regions directly from CT scans. Tissue composition, density, and radiation transport characteristics were assigned to each individual volume element (voxel) automatically depending on its greyscale number and physical location. Both external beam and brachytherapy treatment techniques were simulated using the Monte Carlo radiation transport code MCNP (Monte Carlo N-Particle) version 3A. To obtain a high resolution dose calculation, yet not overly extend computational times, variable voxel sizes have been introduced. In regions of interest where high attention to anatomical detail and dose calculation was required, the voxel dimensions were reduced to a few millimetres. In less important regions that only influence the region of interest via scattered radiation, the voxel dimensions were increased to the scale of centimetres. With the use of relatively old (1991) supercomputing hardware, dose calculations were performed in under 10 hours to a standard deviation of 5% in each voxel with a resolution of a few millimetres- current hardware should substantially improve these figures. It is envisaged that with coupled photon/electron transport incorporated into MCNP version 4A and 4B, conventional photon and electron treatment planning will be undertaken using this technique, in addition to neutron and associated photon dosimetry presented here. PMID- 9745790 TI - Photon buildup in orthovoltage X-ray beams. AB - Orthovoltage x-ray beams exhibit the characteristic of depth dose buildup which is not well described in the literature. The principal reason for this phenomenon is the increase in dose deposited due to electrons set in motion by secondary (Compton) scattered photons within the phantom, as depth is increased until longitudinal equilibrium is reached. This happens within a few millimetres of the surface and has been demonstrated both experimentally and by Monte Carlo methods. The Monte Carlo technique also enabled description of a second order primary dose buildup effect (due to longitudinal electronic disequilibrium) that would be impossible to detect with conventional detectors due to the short range of the electrons. The magnitude of buildup was observed to alter with various combinations of beam parameters. Variations will also occur with detectors used to measure buildup. It is recommended that radiation oncology departments assess this effect in the context of their clinical data in current use to ensure that there are not doses higher than prescribed being applied a few millimetres below the skin surface, especially if data was collected with a thin windowed, parallel plate ionisation chamber and/or that coarse steps for depth dose data collection were used along the beam central axis. PMID- 9745792 TI - Mapping acupuncture points using multi channel device. AB - The practice of acupuncture involves the stimulation of specific points on the skin called 'acupuncture points' which are small regions of local or referred pain that are more sensitive than surrounding tissue. The fact that acupuncture points can be identified subjectively as tender points and are found to have characteristic electrical properties suggest that they are functional entities rather than structural ones. These functional properties are used diagnostically in a clinical setting as pathology in a particular body location has been shown to correlate with increased tenderness and electrical conductivity of the 'corresponding' acupuncture point using electronic 'point locators', which measure the DC resistance of points compared to surrounding skin. Commercially available point locators generally utilize a metal locator probe and an indifferent electrode and are designed to produce an auditory output (usually a high pitched tone) when a point is located. These devices however, are open to criticism. They are unable to control for local variations in skin thickness, surface secretions, or pressure placed on the electrode, and are only able to measure a single point at a time. These make them time consuming to use and subjective to user bias in point selection. Furthermore these devices do not store data and are therefore unsuitable for producing a map of skin resistance, which can be accessed over time. To overcome some of the limitations of currently available single probe devices, we have designed a multi-channel probe capable of measuring and then mapping the skin resistance of multiple points. PMID- 9745791 TI - Paediatric doses from diagnostic radiology in Victoria. AB - This study examines doses to paediatric patients from diagnostic radiology. Measurements were made at 29 hospitals and private radiology practices in the state of Victoria. Entrance skin doses in air were measured for the exposure factors used by hospital radiology departments and private radiology practices for a standard size 1, 5, 10 and 15 year old child, for the following procedures: chest AP/PA, lat; abdomen AP; pelvis AP; lumbar spine AP, lat; and skull AP, lat. There was a large range of doses for each particular procedure and age group. Factors contributing to the range of doses were identified. Guidance levels for paediatric radiology based on the third quartile value of the skin entrance doses have been recommended and are compared with guidance levels. PMID- 9745793 TI - A comparative study of the work involved in measuring profiles using an ion chamber, a linear diode array and film. AB - In this work a method of using Kodak X-Omat V film to measure beam profiles for dynamically wedged fields is presented. Also, the profiles determined by film measurement are compared with those measured with an ion chamber (0.12 cm3 Scanditronix RFA 300 RK) and an array of silicon diodes (11 channel Scanditronix linear diode array). The beam investigated is a 6 MV photon beam from a Varian 2100 C linear accelerator. The geometric method of positioning film and determining the central axis (CAX) position of the beam yielded results which agreed to within 1 mm with the software determined position of the CAX. The profiles measured by film agreed well with the ion chamber measured profiles in terms of overall field size, position, penumbral width, height and position of maximum and profile shape between the 20% dose levels. Film profiles deviated most from ion chamber profiles in the post-penumbra regions. Linear diode array (LDA) measured profiles matched ion chamber profiles in the post-penumbra regions, field size and general profile shape. In the region of maximum dose differences in dose of up to 4% were seen along with horizontal shifts of around 2 mm between LDA and ion chamber profiles. PMID- 9745794 TI - An independent check method of radiotherapy computer plan derived monitor units. AB - With the increasing complexity of radiotherapy computer treatment planning systems (CTPS), verification of the treatments is an important component of a quality assurance program. The radiation dose delivered to a patient is based on treatment machine monitor units (MUs) that are calculated either using data tables or within the CTPS. An independent method for checking radiotherapy MU calculations is proposed. This method involves calculating the dose at a point (usually ICRU reference point) using the derived MUs and independently measured treatment data and comparing to the dose at the same point as predicted by the CTPS. A program was developed using Microsoft Excel to assist in performing this check. The program contains the beam data and various correction factors and calculates the dose after input of the treatment setup parameters. This independent method is used to check all radiotherapy treatment fields at Liverpool Hospital and has found a number of significant errors in the planning process and in the CTPS calculations. PMID- 9745795 TI - A versatile multichannel biotelemetry system. AB - A radiotelemetry system is introduced for analyzing several physiological parameters from free ranging patients within a transmission range of 100 metres. The parameters considered are two independent electromyograms, an electrocardiogram, and a body position indicator. A battery status signal is also transmitted. An equalizer network is utilized to improve the overall frequency response of the system. The composite signal from the equalizer can be recorded by using a tape recorder and demulitiplexed simultaneously. A filter bank is used for demultiplexing. Each filter unit in the bank provides outputs that emphasize and at the same time suppress signals in the same frequency range. The suppressed output supplies the signal to proceeding stages. Hence, an efficient filtering of closely packed multiplexed signals is achieved. The incoming original signals are recovered at outputs of demodulators following the filter bank. The proposed system provides an efficient and cost-effective way of physiological signal detection and storage from free-ranging patients. It is designed using commonly available electronic components. It can be implemented easily, and adapted to other physiological and physical parameters of interest. PMID- 9745796 TI - Cognitive treatment of pathological gamblers. AB - This study evaluates the efficacy of a cognitive treatment for pathological gambling. Five pathological gamblers were treated in a multiple baseline across subjects design. Cognitive correction targeted the erroneous perceptions towards the notion of randomness. Four subjects reported a clinically significant decrease in the urge to gamble, an increase in their perception of control, and no longer met the DSM-IV criteria for pathological gambling. Therapeutic gains were maintained at the 6 month follow-up. Results suggest that cognitive therapy targeting the misconception of the notion of randomness is a promising treatment for pathological gambling, a refractory disorder to most therapeutic interventions. PMID- 9745797 TI - Underpredicted pain disrupts more than correctly predicted pain, but does not hurt more. AB - One of the explanations for the negative effects of underpredicted aversive experiences is that they have more impact than correctly predicted aversive experiences. In a laboratory experiment 40 normal female subjects executed an auditory discrimination task. Subjects were randomly assigned to a correct information condition and an underprediction information condition. After ten trials (baseline) subjects were informed that they would receive some painful (correct prediction) or non-painful tingling (underprediction) stimuli during the discrimination task. Starting just before five of the following 20 discrimination trials, 2 s of painful electrical stimulation was given. Subjects rated sensations and painfulness of the electrical stimulation, subjective anxiety, and degree of distraction from the task, after each pain stimulus. Reaction times of the discrimination task and heart rate were measured. Underprediction information resulted in lower pain ratings, but stronger heart rate responses and higher disruption on the discrimination task, compared to correct information. This suggests that while underpredictions of pain do not hurt more, disruption on primary tasks and physiological impact are higher. Underpredicted pain has more impact than correctly predicted pain, not because it hurts more, but because it conveys inherent danger information. PMID- 9745798 TI - Intrusive memories and depression in cancer patients. AB - Matched samples of depressed and nondepressed cancer patients were interviewed about past life events, particularly experiences of death and illness. They identified and described any spontaneous intrusive visual memories they had experienced in the past week corresponding to these events. About one quarter reported such memories and, as predicted, the majority of intrusive memories concerned illness, injury and death. The mean levels of intrusion and avoidance were equivalent to patients with post-traumatic stress disorder. Consistent with prediction, depressed patients reported significantly more intrusive memories than controls, and described the memories as typically beginning with or being exacerbated by the onset of depression. Greater numbers of intrusive memories were associated with more maladaptive coping, and greater avoidance with deficits in autobiographical memory functioning. PMID- 9745799 TI - Dimensions of perfectionism across the anxiety disorders. AB - To explore the role of perfectionism across anxiety disorders, 175 patients with either panic disorder (PD), obsessive compulsive disorder (OCD), social phobia, or specific phobia, as well as 49 nonclinical volunteers, completed two measures [Frost, R. O., Marten, P., Lahart, C., & Rosenblate, R., (1990). The dimensions of perfectionism. Cognitive Therapy and Research, 14, 449-468; Hewitt, P. L., & Flett, G. L., (1991). Perfectionism in the self and social contexts: Conceptualization, assessment and association with psychopathology. Journal of Personality and Social Psychology, 60, 456-470.] that assess a total of nine different dimensions of perfectionism. Relative to the other groups, social phobia was associated with greater concern about mistakes (CM), doubts about actions (DA), and parental criticism (PC) on one measure and more socially prescribed perfectionism (SP) on the other measure. OCD was associated with elevated DA scores relative to the other groups. PD was associated with moderate elevations on the CM and DA subscales. The remaining dimensions of perfectionism failed to differentiate among groups. The clinical implications of these findings are discussed. PMID- 9745801 TI - Thought control strategies in acute stress disorder. AB - Intrusive trauma-related thoughts and the means to manage them are a central dynamic in posttraumatic stress. Thought control strategies were investigated in survivors of motor vehicle accidents with either acute stress disorder (ASD; n = 20) or no ASD (n = 20). Participants completed the Acute Stress Disorder Interview, the Beck Depression Inventory, the Beck Anxiety Inventory, the Impact of Event Scale, and the Thought Control Questionnaire (TCQ) within four weeks of their accident. Although distraction, social control, and reappraisal were the most common strategies in both groups, ASD participants engaged in punishment and worry more than non-ASD participants. Worry and punishment were also strongly associated with severity of intrusive, avoidance, arousal, and depressive symptoms. Findings are discussed in terms of the role of cognitive strategies in resolving posttraumatic stress. PMID- 9745800 TI - A comparison of psychological and pharmacological treatment of pediatric migraine. AB - A comparison was carried out of the efficacy of psychological and drug treatments for children with migraine. Forty-three children aged between 8 and 16 years (mean age: 11.3 years) who suffered from migraine received either progressive relaxation or cephalic vasomotor feedback, both with stress management training, or metoprolol, a beta-blocker. Psychological treatment was administered in ten sessions lasting six weeks and the drug treatment lasted ten weeks. Relaxation and stress management training reduced the headache index (frequency x intensity of headache episodes), more effectively than metoprolol with cephalic vasomotor feedback and stress management training in between. An overall improvement over time was found with regard to frequency and intensity of headache episodes and analgesics intake. When comparing pre- to post-treatment data, children treated with relaxation training improved significantly in headache frequency and intensity, whereas those treated with cephalic vasomotor feedback improved significantly in headache frequency and duration as well as mood. The clinical improvement was stable at an 8-months follow-up. PMID- 9745802 TI - Cognitive bias in acute stress disorder. AB - Cognitive bias was investigated in survivors of motor vehicle accidents with either acute stress disorder (ASD; n = 17) or no ASD (n = 17). Participants completed the acute stress disorder interview, the Beck depression inventory, the Beck anxiety inventory, the impact of event scale, and a probability questionnaire (PQ) and a cost questionnaire (CQ) within four weeks of their accident. ASD participants exaggerated both the probability of negative events occurring, and the adverse cost of those events more than non-ASD participants. IES-Avoidance scores were the only significant predictors of both PQ and CQ scores. Findings are discussed in terms of the role of cognitive errors in posttraumatic adjustment. PMID- 9745803 TI - Factor analytic investigation of the Illness Attitudes Scale in a chronic pain sample. AB - The Illness Attitudes Scale (IAS) is a self-report instrument comprising nine subscales designed to assess fears, beliefs and attitudes associated with hypochondriasis and abnormal illness behaviour [Kellner (1986). Somatization and hypochondriasis. New York: Praeger.]. The purpose of the present study was to explore the factor structure of the IAS in a chronic pain sample as a preliminary step toward determining the use of this measure in this sample. Hypochondriacal tendencies have been postulated to play a role in maintaining and exacerbating responses to chronic pain and, therefore, appropriate measurement in this sample is important. In the present study, consecutive chronic pain patients presenting to a pain treatment program (N = 198) were administered the IAS. Principal component analysis with oblique (Oblimin) rotation identified that five factors best explain the measure in this population. These factors were (1) fear of illness, (2) effects of symptoms, (3) health habits, (4) disease phobia and conviction and (5) fear of death. The factor structure overlapped to some degree with the scoring of the IAS proposed by Kellner (1986), as well as with the factor structure identified in a non-clinical sample [Ferguson, E. & Daniel, E. (1995). The Illness Attitudes Scale (IAS): a psychometric evaluation on a non clinical population. Personality and Individual Differences, 18, 463-469.]. There were enough discrepancies, however, to suggest an alternative method for scoring the IAS with chronic pain patients. Implications for the use of the measure with chronic pain patients, as well as future research directions for exploring the utility of this measure with chronic pain patients, are discussed. PMID- 9745804 TI - [Acoustic emission analysis of human bones within the scope of clinical diagnosis]. AB - Fractures occurring in human bones produce an acoustic signal, analysis of which permits an evaluation of its source. In the industrial setting acoustic emission analysis (AEA) is used to non-invasively monitor the function of stressed technical systems or parts of systems. During servicing and monitoring of technical systems, acoustic signals emitted by cracks or material deformation are located with the aid of a few acoustic sensors and evaluated for risk identification purposes. With appropriate technology, therefore, both cortical and trabecular bone can be monitored by acoustic emission analysis. A search is currently ongoing for suitable acoustic technology capable of assessing the extent and location of bone defects and predicting associated risks of fractures occurring. In the present study a system for the measurement and analysis of acoustic emission is described which permits the measurement and analysis of acoustic signals obtained from processed and fresh human and porcine femora. In slightly modified form this system was then used to assess the type and extent of acoustic emission obtained from explanted human femora exposed to cyclical torsional loading until fracture occurred. PMID- 9745805 TI - [Examination of trabecular bone structures of the foot skeleton with MRI imaging]. AB - AIMS: To assess trabecular density and structures at various cancellous bone sites in the foot using MR imaging. To compare the MRI findings with the bone mineral density (BMD) values obtained by quantitative computed tomography (QCT) in the lumbar vertebrae. MATERIALS AND METHODS: Bone mineral densities were measured by QCT in lumbar vertebrae 1 to 3 in ten patients with suspected osteoporosis. Sagittal images of the feet were obtained from these patients and also from seven healthy volunteers applying spin-echo and gradient-echo MR techniques. RESULTS: MR imaging revealed differences between patients with osteoporosis and individuals with healthy bones. Comparison of MR and BMD data revealed a significant dependence of MR data on the bone density measured by QCT (alpha < 0.05). In addition, a good correlation (r = 0.73) was found between the results of the different MRI techniques. CONCLUSIONS: The MR imaging techniques investigated offer an interesting tool for the non-invasive assessment of bony structures, at least at peripheral locations with yellow bone marrow. To enable reliable application in the clinical routine setting, additional studies are required to identify suitable standardized methods and establish normal ranges. PMID- 9745806 TI - [Primary stability of 2 acetabulum roof cups and an acetabulum reinforcement cup. Effect of osseous defects]. AB - The aim of this experimental study was to analyse the effects of implant design and bone stock defects on the primary stability of three different acetabular components. Fresh frozen human pelves were employed for the investigation. Muller and Ganz rings and a Burch-Schneider cage (Protek, Munsingen, Switzerland) were fixed with screws in 6 normal acetabula using standard techniques, and in 30 acetabula previously prepared with 5 different segmental bone stock defects. A servohydraulic testing machine (Instron, Canton, USA) was used for the investigation. Three electromagnetic displacement transducers (Micro-Epsilon, Ortenburg, Germany) were placed in the three main quadrants of the acetabular rim to detect implant micromotion, which reflects stability. Displacement was recorded during 20 consecutive cycles under loads of up to 2354 N. All implants were stable (< 150 microns) in all quadrants of normal acetabula and also in those with ectatic, protrusive and ventral defects. There was no statistically significant difference in the results between Muller and Ganz rings. Displacement of more than 150 microns was observed in acetabula with cranial or dorsal defects. The cage was stable under all defect conditions. The reinforcement implants showed low displacement rates in most of the acetabular bony defects. Stability is a function of the area of surface contact between prosthesis and bone. PMID- 9745807 TI - [Percutaneous bipolar discectomy. Technical principles and initial results with an in vitro model]. AB - In the present study, the effect of bipolar radio-frequency thermocoagulation of intervertebral discs was evaluated in an in vitro model. In fresh human cadaveric thoracic and lumbar motion segments, we applied radio-frequency energy to the intervertebral disc via a variety of different bipolar HF electrodes, while simultaneously recording the temperature of the dorsal longitudinal ligament. All the segments were weighed in a standardised manner prior to and after the procedure. It was found that a combination of coagulation and vaporisation was most suitable for working on the disc. Applying energy for 300 secs resulted in an average weight loss ranging, electrode-dependently, from 0.3 to 0.7 grams, the mean temperature increase in the longitudinal ligament was 0.1 degree C per min. CLINICAL RELEVANCE: Our experimental data suggest that PBD may be used for the same indications as the laser in the treatment of disc disease. PMID- 9745809 TI - Comparative study of two non-steroidal anti-inflammatory eyedrops, 0.1% indomethacin versus 0.1% diclofenac in pain control post photorefractive keratectomy. AB - We evaluated the efficacy and safety of 0.1% indomethacin and 0.1% diclofenac solutions, in controlling pain post excimer laser photorefractive keratectomy (PRK). After written consent, 61 informed patients (23 males, 38 females; mean age = 33.5 +/- 8.4 yrs) were enrolled in a double-masked, randomized, comparative study and assigned to either indomethacin or diclofenac treatment. Subjective preoperative evaluation of individual susceptibility to pain evoked by topical application of 1% tetracaine vs saline served as reference for further post operative pain measurement using a visual analog rating scale. Ocular and cephalic pain, itching, foreign body sensation, insomnia, photophobia, blepharospasm as well as systemic analgesic medication and alcohol intake were monitored for 3 days following photoablation as well as the re-epithelialization process. Both solutions significantly reduced pain on the first day following excimer laser PRK, and this activity was maintained until the end of the observation period. At Day 0 the first measure of pain level was slightly higher in the indomethacin group (p < 0.05) and could be related to a possible anaesthetic effect of Diclofenac. During the follow-up the oral intake of analgesics was higher in the diclofenac group, however this difference was not significant. Wound healing rate was not affected by indomethacin or diclofenac administration. These data suggest that both 0.1% indomethacin and 0.1% diclofenac ophthalmic solutions may help to control the pain induced by excimer laser PRK without any deleterious effect on corneal wound healing. PMID- 9745808 TI - [Algorithm for automatic endocardium identification in digital echocardiography image sequences]. AB - The present paper describes an automated procedure for the detection of left ventricular internal wall edges in digital echocardiographic image sequences. The proposed procedure is divided into three steps and programmed in C/UNIX. It includes the use of a specially designed, application-specific adaptive spatio temporal filter for noise reduction in echocardiographic image sequences, local 3 D histogram equalization for augmented contrast, and segmentation of the left ventricle using a regional growth method. When designing the adaptive spatio temporal filter, we took into account the fact that the background noise is correlated in tangential orientation due to beam deflection at interfaces characterized by a large impedance "jump". Using the specially designed filter, the background noise is successfully reduced without degrading the ventricular contours. The simulation results presented highlight the performance of the proposed method in an exemplary manner. PMID- 9745810 TI - Glaucoma in Congo. AB - OBJECTIVE: To determine frequencies of various types of glaucoma in an urban community of Congo. METHODS: The records of 176 patients with the diagnosis of glaucoma examined between 1991 and 1995 at a non-university center were reviewed. A complete ophthalmological examination was performed in each patient including best visual acuity, refraction, slit-lamp examination, ophthalmology and intraocular pressure measurements. Gonioscopy and visual field examination were performed when needed. RESULTS: The frequency distribution of the various subtypes of glaucoma was: open-angle glaucoma (72.2%), aphakic glaucoma (9%), uveitis glaucoma (6%), exfoliative glaucoma (3%), glaucoma secondary to ocular trauma (2.2%), cataract glaucoma (1.7%), normal tension glaucoma (1.7%), congenital glaucoma (1.7%), neovascular glaucoma (1.3%), pigmentary glaucoma (0.6%), corticosteroid-induced glaucoma (0.6%), and glaucoma associated with retinoblastoma (0.6%); CONCLUSION: Despite the limitations inherent of a retrospective review of records, our study provides an indication of the prevalence of various types of glaucoma in a urban community of Congo. This study confirmed also the rarity of primary angle-closure glaucoma in the black. PMID- 9745811 TI - Risk factors for open-angle glaucoma in 260 black subjects in Congo. AB - OBJECTIVE: To identify possible risk factors for open-angle glaucoma (OAG) among 260 black subjects in Kinshasa (Congo). METHODS: Between May 14, 1996 and June 14, 1996, 260 persons, aged 24-60 years old, working at a factory in Kinshasa, have been examined. Twenty-two patients with OAG and 238 controls were identified. Data were obtained through interviews and clinical examinations. OAG was defined by the presence of both optic disc damage and characteristic visual field defects. RESULTS: The frequency of OAG was 8.5%. Associations were found with intraocular pressure greater than 21 mmHg (Odds ratio (OR) = 119.4, 95% confidence interval (CI), 17.62 to 4960.64), current cigarette smoking (OR = 2.77, 95% CI, 1.04 to 7.34) and Mongo ethnic subgroup (OR = 3.195, 95% CI, 0.93 to 9.57). CONCLUSION: Mongo ethnic subgroup seemed to be another risk factor for OAG in Congo. PMID- 9745812 TI - Spontaneous evolution of stage I and II macular holes. AB - To ascertain the natural outcome of stage I impending and stage II macular holes, 28 eyes (26 patients) were reviewed and followed up for an average of 23 months. Of the 21 stage I lesions, 10 (47.6%) progressed and the others regressed. In the stage II group (9 eyes), 5 lesions (55.5%) progressed, 2 (22.2%) regressed and 2 (22.2%) remained stable during a follow-up period of 7.75 months. Patients with stage II macular holes seem to be more likely to benefit from surgery than patients with stage I lesions. However, even in stage II macular holes, regression may be observed and the possible benefit from surgery must be weighed against the known complications and risk factors. PMID- 9745814 TI - [Iris metastases in two patients, important for the diagnosis and treatment of the primary tumor]. AB - Two cases of cancer with secondary metastases to the iris were examined. In the first case, the work-up performed after appearance of the iris lesion led to the discovery of a general spread from the already known and treated primary tumor which was an oat cell carcinoma of the lung. PMID- 9745813 TI - Indocyanine green angiographic findings in multifocal choroidopathies. AB - With high definition videoangiography (TOPCON IMAGEnet H1024) the Authors studied 41 patients affected by multifocal choroidopathies (MC) (68 eyes with ophthalmoscopic or indocyanine green angiographic evidences): 29 females and 12 males; age 21-51 years with a follow up of 6-29 months. In the light of the evidence provided by FA and ICG the Authors present a classification of MC in three stages: Stage 1 of subclinical choroidal activity (5 eyes) characterised by the presence of hypofluorescent or hyperfluorescent spots visible only in the late phases of ICGA; stage 2 of clinically evident choroidal activity (45 eyes) in FA the spots are hypofluorescent in the early phases and hyperfluorescent with a slight diffusion in the late phases, in ICGA either hypofluorescent spots or less frequently hyperfluorescent spots and choroidal permeability alterations can be observed; stage 3 or healed stage (18 eyes) in FA the spots are hyperfluorescent without late leakage, in ICGA hypofluorescence can be observed during all angiographic phases. In 5 patients in stage 1 of subclinical activity, a systemic steroid therapy induced a regression of the hypofluorescent spots in ICGA, in 2 cases the regression of hyperfluorescent spots in ICGA was observed after systemic antibiotic therapy. The authors underline that ICGA could be a particularly useful tool for an early diagnosis and clinical monitoring of MC. PMID- 9745815 TI - [Acute pancreatitis, papillary coloboma and serous macular detachment]. AB - A serous macular detachment in one eye with spontaneous flattening is seen twice during acute pancreatitis in a man aged thirty-four years who has a pancreatic malformation and an optic disc coloboma of this eye. Fluorescein angiography discloses the classic sequence of an optic disc pit complicated by maculopathy. Vascular leakage attributed to acute pancreatitis is also seen in the macula. Every time, both retinal detachment and vascular leakage have disappeared in few days. The mechanism responsible for the development of the macular detachment in this case is discussed. PMID- 9745816 TI - [Precocious exudative retinal detachment after surgery of the macular hole]. AB - The retrospective study of our patients operated on for an idiopathic macular hole shows 2 cases of presumed exudative retinal detachment appearing 24 to 36 hours after surgery. This complication, to our knowledge, has never been reported. We describe these 2 cases and discuss on one hand, the arguments for an exudative etiology of the retinal detachment, and on the other hand, the possible causes of this unusual complication. PMID- 9745817 TI - [Laser treatment of detachments of the retinal pigment epithelium associated with sub-retinal neovascularization in the context of age-related macular degeneration: retrospective study]. AB - Pigment Epithelial Detachment (PED) associated with subretinal new vessels (SRNV) is a particular aspect of Age Related Macular Degeneration (ARMD). We retrospectively analysed the results of dye laser photocoagulation in 63 eyes of 56 patients with vascularised PED. We photocoagulated in a confluent manner the presumed zones of SRNV, detected by fluorescein angiography and three-mirrors lens examination. In most cases, the SRNV were of the occult type. In 89% of the treated eyes we obtained a flattening of the PED and the visual acuity was stabilized or ameliorated in 66% of the cases after a mean follow up of 29 months. This final visual acuity was better or equal to 1/10 in 64% of the cases and superior or equal to 5/10 in 46% of the cases. Subfoveal SRNV, initial visual acuity of less than 1/10, and persistence or recurrence of the PED after treatment were of bad prognosis. However, recurrence of the SRNV was not necessarily of bad prognosis if it could be retreated. Treatment of interpapillomacular SRNV had the best prognosis. Laser photocoagulation can be beneficial in well selected patients with vascularised PED. PMID- 9745818 TI - Retinal capillary hemangioma: von Hippel (-Lindau) disease. AB - The clinical manifestations of von Hippel and von Hippel-Lindau's disease are illustrated. Four cases are presented with special emphasis on the angiographic characteristics, the evolution and the complications of the retinal capillary hemangiomas. Special attention is given to fundoscopy, fluorescein and indocyanine green angiography and to the systemic manifestations. We summarize new molecular genetic insights. The importance of systemic and familial evaluation is stressed. Retinal angiomatosis is a clinical diagnosis but the angiography has an adjuvant value. PMID- 9745819 TI - The Hermansky-Pudlak syndrome. AB - The Hermansky-Pudlak syndrome (HPS) associates oculocutaneous albinism with a haemorrhagic diathesis and the accumulation of ceroid-like material in different tissues. HPS is not an uncommon type of albinism as it was diagnosed in 13.5% (8/59) of our autosomal recessive albinos. These eight patients were evaluated ophthalmologically and haematologically. Apart from the symptoms caused by the albinism, accompanying signs vary from ecchymoses to life threatening haemorrhages and death by associated restrictive lung disease. Recognition of this syndrome by the ophthalmologist can be of major importance in this serious and eventually fatal disorder. PMID- 9745820 TI - [Retinal occlusive vasculopathy and primary anti-phospholipid syndrome: when to evoke it?]. AB - A severe case of retinal vascular occlusion in a 31 year old white female is reported. Its origin is attributed to the primary antiphospholipid syndrome. The diagnostic criteria and the treatment of the primary antiphospholipid syndrome are revised and discussed. When the etiology of a retinal vascular occlusion is not assessed by the initial work-up, especially if the initial presentation consists of a severe occlusive disease, antiphospholipid antibodies should be checked. PMID- 9745821 TI - [Clinical aspects of three cases of bilateral retinoblastoma]. PMID- 9745822 TI - [Cesarean, result of an exploration of a loss of visual acuity]. AB - Toxaemia of pregnancy associating hypertension and proteinuria can cause maternal and ocular complications. Maternal ocular involvement is classically described with loss of visual acuity due to serous retinal detachment. We report a case of a 31 year old woman who just complained of severe deterioration of visual acuity. During her stay at the hospital we discovered a pregnancy complicated by a HELLP syndrome. Resturation of vision has been obtained after fetal expulsion and medical treatment against hypertension. PMID- 9745823 TI - [Distinguishing clinical manifestation of panuveitis with a cytomegalovirus retinopathy in a grafted person]. AB - PURPOSE: Unusual description of an uveitis with a cytomegalovirus retinitis in a grafted patient. MATERIAL ET METHOD: A 49 year-old white male had to receive an hepatic transplant. Six month later, a cytomegalovirus chorioretinitis was discovered with an important uveitis. CONCLUSION: Two to five percent of cytomegalovirus retinitis were found in grafted patients. No patient was seen with a significant uveal reaction. In the article will be discussed the evolution, the diagnosis and the treatment of this case report. PMID- 9745824 TI - [Impact of anti-retroviral treatment with protease inhibitors on the evolution of cytomegalovirus retinitis in the patient carrying acquired immunodeficiency syndrome]. AB - Up to now cytomegalovirus retinitis (CMVR) recurrence in AIDS patients was considered to be very high, even on maintenance therapy. Protease inhibitors (PI) are antiretroviral molecules which, with more efficacity than reverse transcriptase inhibitors, decrease viral charge and increase CD4's count aswell as survival. We analysed the impact of PI on CMVR evolution in a retrospective review of 18 patients with CMVR on maintenance therapy and PI treatment. In a first group, 13 patients started PI some time after CMVR diagnosis (median CD4 = 9/mm3). A second group of 5 patients developed CMVR (median CD4 = 63/mm3) after initiation of PI. In the first group, incidence of CMVR recurrences/1000 patients days was 6,45 (2323 patients days of follow-up (PDFU) before starting PI and 3,44 (4066 PDFU) after starting PI. In this group, during the follow-up's period of CD4's count inferior to 75/mm3, incidence of CMVR is 6,84/1000 patients days and becomes 0,86/1000 patients days during the follow-up's period of CD4's count superior to 75/mm3. In the second group, incidence of CMVR was 0/1000 patients days (1972 PDFU). In summary, incidence of CMVR decreases with PI's treatment. Interruption of CMVR maintenance therapy could be considered in patients with CD4's count higher than 75/mm3. PMID- 9745825 TI - [Colored filters in the treatment of severe photosensitivities. Introduction to a pilot study]. AB - Among photosensitive patients, the use of colored filters sometimes leads to a clear improvement of comfort. After a general review of theory, we start on the way we take care of these patients and the way filters are fitted and made. We only present here a preliminary work. A subsequent study or accurate informations of colored filters will be published later. The aim is to set up a methodic procedure in order to find the best selection of the recommended spectral curve and the best way to follow the progress of the patient as regards the wearing and the effect of the filter. An overview of our cases is presented. PMID- 9745826 TI - [Treatment of glaucoma with carbonic anhydrase inhibitors in eyewash: medium term retrospective experience with dorzolamide]. AB - AIM: Clinical evaluation of the efficacy and innocuity of dorzolamide, the first new born topical carbonic anhydrase inhibitor. MATERIALS AND METHODS: Our retrospective analysis included 162 patients (76 men, 86 women) who were exclusively followed in our department and treated between June 96 and August 97 with dorzolamide (Trusopt 2%) administered in a minority of patients (n = 13) as the only hypotensive medication and in combination with other hypotensive drugs and/or as a substitute for acetazolamide or for miotics in the others (add on, n = 149). The mean age of the patients was 65.2 +/- 15.6 and the average follow up was 9.8 +/- 5.4 months. Dorzolamide was prescribed in various forms of glaucoma and ocular hypertension, mostly in POAG (n = 137). In bilateral treatments (n = 116/162), the eye with the most severe defect and/or the highest IOP was considered. IOP was checked along the other criteria generally followed in glaucoma patients at 1 and 3 months following the instauration of dorzolamide and then trimestrially. We considered as the initial IOP a mean value of the 2 most recent successive IOP values before instauration of dorzolamide realized at the same daytime. Patients with laser trabeculoplasty or cataract surgery during the dorzolamide treatment were disguarded. RESULTS: All patients considered, the mean IOP reduction was from 21.5 mm Hg before to 19.3 mm Hg with dorzolamide at the last control. The mean IOP decrease was statistically significant and stable at all controls. The mean IOP reduction was 18.4% (4.2 mm Hg) in the monotherapy group and 12.5% (3 mm Hg) in the add on group. There were 15.3% non responders (mean IOP reduction < half of the average IOP decrease observed all patients confounded) in the monotherapy group and 26% in the add on group. Local tolerance of dorzolamide was excellent in 84%. About 15% of patients (25/162) complained of systemic side effects mostly comparable to the side effects of oral IAC. Treatment was stopped in 57 patients (35.2%) primarily due to allergic blepharoconjunctivitis (n = 12 or 7.4%), general intolerance (n = 14 or 8.6%) and insufficient IOP reduction (n = 26 or 16%). CONCLUSIONS: Dorzolamide represents in our experience an effective ocular hypotensive drug but its systemic tolerance profile seems less promising than described in the literature. PMID- 9745827 TI - [The Baerveldt implant in refractory glaucoma: clinial results]. AB - 25 Baerveldt glaucoma implants were placed in 22 patients, between 1992 and 1997. The indications were: 14 cases of neovascular glaucoma, 4 congenital glaucomas, 3 glaucomas due to intraocular silicone oil and 4 various glaucomas. The mean preoperative intraocular pressure was 46 mmHg (26-77) on a mean of 2.1 glaucoma medications. The postoperative mean intraocular pressure was 17 mmHg (5-50) on a mean of 0.5 glaucoma medications with a mean follow-up of 16 months (1 week-48 months). The final intraocular pressure was less than or equal to 20 mmHg in 20 of the 25 eyes. The final visual acuity improved or remained the same in 10 of the 25 cases and worsened in 11 cases (4 cases undetermined). The most frequently observed complications were: hyphema, choroidal effusion and corneal edema. Our results, in terms of intraocular pressure control, with the Baerveldt implant in patients with complicated glaucomas appear to be satisfactory. The use of this implant could be extended. PMID- 9745828 TI - [Transvitreal endocryoapplication of the ciliary body]. PMID- 9745829 TI - Differences in the trabecular meshwork between Belgian and Congolese patients with open-angle glaucoma. AB - PURPOSE: To study the alterations of the trabecular meshwork of Belgian and Congolese patients with open angle glaucoma (OAG). METHODS: A trabeculectomy was performed in 27 OAG patients from Belgium and 24 from Congo; the trabecular specimens were fixed, embedded, stained, and studied by light microscopy. These specimens were compared with the trabecular meshworks from 5 Belgian non glaucomatous eyes. RESULTS: The mean number (+/- standard deviation) of trabecular cells per field was 69 (+/- 10) in Belgian normal eyes, 34 (+/- 17) in Belgian OAG patients, and 10 (+/- 6) in Congolese OAG patients. The trabecular meshwork was collapsed and Schlemm's canal was closed in the cases with a diminished number of cells. Pigmentation was present in 79% of the Congolese and in 35% of the Belgian specimens. CONCLUSION: The trabecular meshwork from OAG patients has a lower cellularity than normal, and the effect is much more pronounced in the trabecular meshwork from Congolese patients. This may be a possible explanation for the racial differences in OAG and the more severe forms of glaucoma in black people. PMID- 9745830 TI - Interpretation of automated perimetry. AB - This paper discusses basic principles of visual field interpretation. Only the two most popular perimeters, the Octopus and the Humphrey Field Analyzer, were considered. Choices of tests and strategies, software to assist interpretation (with emphasis on glaucoma progression), and future trends in visual field testing are covered. PMID- 9745831 TI - The differential diagnosis of posterior uveitis. AB - The diagnosis of posterior uveitis can be established in most cases on the basis of (1) morphology of the lesion, (2) the mode of onset and course of the disease, and (3) the association with other systemic diseases. The differential diagnosis of posterior uveitis include the following entities: (1) the viral infections, (2) the bacterial infections, (3) the parasitic infections, (4) the fungal infections, (5) the autoimmune diseases, and (6) the diseases of unknown origin. PMID- 9745832 TI - Salt and hypertension: a race-related phenomenon? PMID- 9745833 TI - Salt taste sensitivity and blood pressure in adolescent school children in southern Nigeria. AB - This cross-sectional study was undertaken to observe the relationship between taste recognition threshold for NaCl (salt taste sensitivity) and blood pressure (BP) prior to the onset of clinical hypertension. The study involved 160 girls and 159 boys aged ten through seventeen years. Three hundred and nineteen (35%) of the total population were relatively insensitive to NaCl taste (defined as threshold value > or = 60mM NaCl). In this, there was significant gender difference (chi 2 = 9.66, df = 2, p = 0.022) in the distribution of salt taste sensitivity, with more boys than girls having higher threshold values (> or = 60mM NaCl). In addition, a highly significant (p = 0.0103) but weak association (r = 0.1439) between salt taste sensitivity, and systolic BP was evident after adjustment for other confounding variables. This finding appears fairly specific for NaCl sensitivity, as no systematic relationship between any of the individuals parameters of BP (that is systolic, diastolic and mean arterial pressures) and threshold values for sucrose, urea and HCl was apparent. Overall, however, the bulk of the data does not support a major role for NaCl taste sensitivity in the determination of BP levels in this group of Nigerian adolescents. Rather, height and gender were the most important factors that influenced BP. PMID- 9745834 TI - Biliary excretion in persons with low blood glutathione levels. AB - A study to investigate the association between blood glutathione (GSH) levels and biliary excretory status was conducted in apparently healthy Ghanaian subjects without frank biliary disease and anaemia. The results showed that, in adults (mean age: 38.5 years) and children (mean age: 13.0 years), plasma conjugated bilirubin is inversely correlated with blood GS (respective site r = -0.524, p < 0.011 and -0.395, p < 0.005). Persons with elevated plasma conjugated bilirubin compared to controls (mean: 6.0 versus 2.5 umol/L, p < 0.001) also exhibited low blood GSH values (3.5 versus 4.2 umol/gHb, p < 0.029). Malaria parasites with counts up to 2,453 parasites/ul blood had no effect on the obtained data. The results suggest that low blood GSH levels may be relevant to delays in biliary excretion of conjugated toxins from the liver, as exemplified by the rise in conjugated bilirubin levels in the plasma, and predispose liver cells to increased oxidant state and damage. PMID- 9745835 TI - Impotence in Ethiopian diabetic men. AB - Prevalence of impotence was assessed among 292 consecutive diabetic men attending the Tikur Anbessa Hospital, diabetic clinic. The mean age was 41.4 +/- 15.5 years (range 18-86 years). One hundred and forty nine (51.6%) were type I and 143 (49%) were type II patients. The mean duration of diabetes was 9.9 +/- 6.7 years and 37.7% had known long term diabetic complications. The overall prevalence of impotence was 48.7%. The mean duration of impotence was 3.5 +/- 3.4 years. In the majority of the cases, impotence started after diagnosis of diabetes mellitus. Many of the patients (79.1%) had never complained to physicians and 59.2% of the patients did not know that impotence is a complication of diabetes mellitus. All except 10 patients (7.5%) had libido. Impotence is significantly higher in type II as compared to type I patients (94/143 versus 40/132, p < 0.001) and in patients with complications than without (76/104 versus 54/159, p < 0.001). The mean duration of diabetes mellitus is significantly higher in patients with impotence than without impotence (12.3 years versus 8.1 years, p < 0.001). We conclude that impotence is a common and significant problem in our diabetic men and we recommend further study to assess its social and psychiatric impact. PMID- 9745836 TI - Profile of diabetic Omani pilgrims to Mecca. AB - The annual pilgrimage to Makkah (Mecca), Hajj, is a very stressful endeavour and requires strenuous physical effort, especially for the diabetic, the elderly and persons with other chronic illnesses. To identify the complications and to assess the needs of the Omani diabetics during Hajj (DOH), a special diabetes clinic was established in the camping site of Omani pilgrims (Hajjees) in Mina, where all Omani Hajjees convene for three days. The socio-demographic characteristics, the diabetes profile and the knowledge about complications of diabetes of all DOH were ascertained; their random blood sugar (RBS) was tested. Of 10,800 Omani who performed the Hajj in 1996, the 169 Hajjees with diabetes mellitus (prevalence rate 16 per 1000) included four per cent insulin dependent (IDDM), seven per cent on dietary control, and 89% on oral hypoglycaemic agents. Almost all DOH (98%) were medically examined before their departure for Hajj. All Hajjees with IDDM and 96% on oral hypoglycaemic agents brought their medicines with them. During the Hajj period, 2.4% of DOH had RBS < 75 mg/dl, 14% 75-110 mg/dl, and 49% were hyperglycaemic (RBS > 200 mg/dL). About half of the DOH (48%) knew the clinical presentation of hyperglycaemia, a fourth (24%) about symptoms of hypoglycaemia. Only 9.5% were trained to test themselves for blood sugar. The median age of DOH was 54 years (inter-quartile range 50-62). Some 7.5% females and 4.9% of males were obese (body mass index > 30). Forty seven (28%) of the DOH had other coronary heart diseases, hypertension or both. DOH moved between Holy places (four journeys; 5-15 km long) on foot (40%), by car or bus (31%), or both (29%). All DOH except one were not wearing protective shoes, 70% did not have identification wrist bands that show their diabetic status and regimen for treatment. Four per cent lost their way during Hajj, four per cent suffered from heat exhaustion, three per cent had cut wounds, 1.2% had pneumonia, and two per cent went into coma. There is a need for a special health education programme and for special services for the diabetics during Hajj. Hajjees should learn about symptoms and signs of hypoglycaemia, were protective shoes and identifying wrist bands. Specialised services for the diabetics would alleviate a lot of the stress during Hajj among the diabetics. PMID- 9745837 TI - Prevalence of food-borne pathogens and growth potential of Salmonella in weaning foods from Addis Ababa, Ethiopia. AB - One hundred samples of ready-to-consume feeding bottle contents from outpatient infants were examined for the presence of Salmonella, Staphylococcus aureus and Bacillus cereus. Nearly all samples had very high gross bacterial contamination. Three Salmonella isolates were encountered from bottle contents made of cow's milk and gruel made from cereal blend. All belonged to group D. Of the 108, isolates 67 were Staph. aureus and 38 were B. cereus. The potential of Salmonella spp to grow in weaning foods was also determined on one common factory-produced infant food and one home-made infant food. In both items, Salmonella increased by approximately 4 log units within eight hours and reached counts as high as log 8 cfu/ml within twelve hours. PMID- 9745838 TI - Assessment of the Rose-Bengal plate test for the diagnosis of human brucellosis in health facilities in Narok district, Kenya. AB - The Rose-Bengal plate test (RBPT) was performed on 488 patients with flu-like symptoms from Narok district. There was poor agreement between RBPT results from four health facilities in Narok and from the central veterinary laboratory (CVL). Agreement was poorer for the three rural dispensaries than for the District Hospital. On the other hand, for tests conducted at the CVL, there was good agreement between RBPT, serum agglutination test (SAT) and complement fixation test (CFT) results, indicating that all these tests were probably performing well. Better training and quality control and the use of white rather than a clear background surface for judging agglutination results are recommended to improve the performance of test results in Narok District health facilities. PMID- 9745839 TI - Late presentation of laryngeal and nasopharyngeal cancer in Kenyatta National Hospital. AB - The clinical stage at presentation of laryngeal and pharyngeal cancer is an important determinant of survival. Fifty six patients admitted at Kenyatta National Hospital with nasopharyngeal and laryngeal carcinoma were reviewed to determine the period of delay from onset of illness to the first otolaryngologic appointment at the hospital, their clinical features and the tumour stage at presentation. All cases of nasopharyngeal carcinoma had cervical lymphadenopathy, 70.6% being N3 status while 57.8% of laryngeal carcinoma cases underwent emergency preoperative tracheostomy due to bulky obstructive tumour. On the whole, 96.4% of the patients presented with advanced (stage 3 or 4) head and neck carcinomas. The average period of delay between the first medical attention at a primary health care facility and the first appointment at the national hospital was 8.7 months. The study suggests that this long delay was due to inherent inefficiency in the referral system and was a major contributing factor to the advanced stage at presentation. PMID- 9745840 TI - Spinal block caesarean section in parturients with pregnancy-induced hypertension. AB - The objective of this work was to determine whether parturients with pregnancy induced hypertension (PIH) are at higher risk of post-spinal hypotension at caesarean section. This was an observational study of 24 women with PIH undergoing caesarean section under spinal analgesia with 0.5% hyperbaric bupivacaine, compared with 24 matched normotensive parturients receiving a spinal block for caesarean section. The mean intra-operative systolic arterial pressure (SAP) was similar with and without PIH (p = 0.38). The mean percentage decrease in SAP of baseline was more with PIH (16.2%) than in the controls (0.5%) (p < 0.001). The number of episodes of severe hypotension (SAP decrease to < or = 80% of baseline and < 90 mmHg) (p = 0.80) as well as the magnitude (p = 0.31) of severe hypotension was similar in both groups. There was no difference in the evolution of diastolic arterial pressure and maternal pulse rate between cases and controls. Maximum levels of upper sensory blockade were similar. Foetal and maternal outcome was similar with and without PIH. The decrease in SAP is less on an absolute scale but more on a percentile basis with PIH at caesarean section under spinal analgesia than in normotensive patients. The difference, however, is not clinically sufficient to discourage spinal analgesia for caesarean section with a low dose (1.5 ml, 7.5 mg) of 0.5% hyperbaric bupivacaine in parturients with PIH. PMID- 9745841 TI - Health-seeking behaviour of patients with sexually transmitted diseases in Zambia. AB - The aim of this paper is to describe health-seeking behaviour, time with symptoms and sexual activity during symptom period among patients attending the public health sector in urban and rural Zambia for treatment of an STD. The study was conducted at two urban health centres and at one rural mission hospital during four months in 1994 and 1995. Four hundred and seventy nine patients seeking health care for STD symptoms were interviewed. The patients had experienced STD symptoms for one to two weeks before they came to the clinic. During this period two thirds in the urban and one third in the rural setting had had sex. Sixty per cent of the patients in the urban and 50% in the rural setting had taken some kind of medicine before they came to the clinic. More people had used modern compared to traditional medicine, especially in the urban area. Market places, other clinics and doctors, friends, and relatives were common treatment sources. Ten per cent had received medicine from a traditional healer. Thus, a majority of the patients had received medication from other sources before they came to the clinic. Sex during periods with STD symptoms was common. This has serious implications for STD as well as HIV transmission. PMID- 9745842 TI - Evaluation of community compliance with annual ivermectin treatment of onchocerciasis in Patigi, Nigeria. AB - A low community acceptance and compliance with annual ivermectin treatment was recorded in Patigi, Nigeria where ivermectin is being distributed centrally in designated centres for the fifth time. The average community acceptance rate (ACAR) for the treatment period was 53.48% while the community compliance rate (CCR) of 1.9% of the total subjects interviewed was recorded. Absenteeism from treatment was a major reason for non-treatment (average, 34.6%). Inadequate and poor mobilisation was identified as a major reason for the low ivermectin acceptance and compliance with treatment in this community. An intensive mobilisation with health education messages with the inclusion of religious/traditional institutions is advocated. PMID- 9745843 TI - Post operative nausea and vomiting in Nigerians. AB - A retrospective survey of postoperative nausea and vomiting (PONV) in the recovery room over a five year period was conducted, followed by a prospective study of 200 adult patients to estimate the incidence and predisposing factors to nausea and vomiting during the first 24 hours after anaesthesia and surgery in Nigerians. In the retrospective study only records of 61 patients (0.79%) out of the 7714 post anaesthetic recovery room charts reviewed revealed documentation of vomiting. These were 20 males (32.8%) and 41 females (67.2%). In the prospective study, the incidence of post operative nausea and vomiting within twenty four hours of surgery was 41.6% and 19.6%, respectively. But only two out of 39 patients (one per cent) vomited within the first three hours in postoperative period. The frequency of vomiting varied from one to 15 times and women had significantly more emetic symptoms than men (p < 0.05). Preoperative administration of pethidine and morphine was associated with postoperative nausea and vomiting. It is suggested that Nigerian women should be considered for prophylactic anti-emetic therapy, especially when narcotic analgesic are to be employed in their anaesthetic management. PMID- 9745844 TI - Clinical presentation and CAT scan findings in mycetoma of the head. AB - Nine cases comprising seven males and two females with mycetoma of the cranium were studied between January 1990 and June 1997. Streptomyces somaliensis was the most common causative organism. The source of the infection was thought to be known in only three cases. The common mode of presentation was headache and scalp swelling. The next common presentation was epilepsy. Other focal neurological disorders also occur. CT scan findings of the cranium showed osteosclerotic rather than osteolytic changes. PMID- 9745845 TI - Listeria septicaemia and meningitis in a neonate: case report. AB - We report the first case of neonatal septicaemia and meningitis in Trinidad due to Listeria three days after blood transfusion. It is important to be aware of the organism in foods and patients. Modern methods of isolation and identification will be of invaluable assistance in future recognition of the organism. PMID- 9745846 TI - Pseudo hypoparathyroidism in a Saudi patient with Klinefelter's syndrome: case report. PMID- 9745847 TI - Re: Health in Africa: what of the future? PMID- 9745848 TI - Corticosteroid therapy in glomerulonephritis. PMID- 9745849 TI - Glycogen and liver dysfunction in anorexia nervosa. PMID- 9745850 TI - Efficacy and limitation of F-18-fluorodeoxyglucose positron emission tomography during fasting to assess myocardial viability in the acute phase of myocardial infarction. AB - OBJECTIVE: The present study was designed to determine the ability of positron emission tomography (PET) to assess myocardial viability and ischemia in acute myocardial infarction (MI) after reperfusion therapy (thrombolysis and/or coronary angioplasty). METHODS: PET with fluorine-18-labeled fluorodeoxyglucose (FDG) under fasting conditions and thallium-201 single-photon emission computed tomography (TL) were analyzed in 21 patients one week following MI. Myocardial viability was also assessed by regional wall motion using serial analysis of 2-D echocardiography or left ventriculography. RESULTS: Marked uptake of FDG together with a residual perfusion defect were observed in the infarct region in all patients one week post MI. However, in 9 of the 21 patients, the infarct-related coronary artery had no significant stenosis after reperfusion therapy and remained patent in one month post MI suggesting no myocardial ischemia. In contrast, in 4 of the 21 patients the regional wall motion was akinetic and there was a complete defect observed with TL imaging at one month post MI, indicating no viability in the infarct region. CONCLUSIONS: PET using fasting FDG at one week post MI had a limitation to predict myocardial viability or ischemia. PMID- 9745851 TI - Three-dimensional anisotropy contrast (3DAC) magnetic resonance imaging of the human brain: application to assess Wallerian degeneration. AB - Three-dimensional anisotropy contrast (3DAC) magnetic resonance imaging is a new algorithm for the treatment of apparent diffusion tensor using the three primary colors. To determine if 3DAC has a clinical application for human brain, six normal volunteers and twenty patients with supratentorial cerebrovascular accidents were examined using clinical magnetic resonance imaging (MRI), and the changes in the 3DAC images associated with Wallerian degeneration of the pyramidal tract were evaluated. The 3DAC images exhibited impressive anatomical resolution. In all chronic stage patients with hemiparesis, the colors in the pyramidal tract were faded. Patients examined during the acute stage who later recovered from hemiparesis had no visible changes of the 3DAC image, whereas patients who recovered poorly showed distinct color fading in the pyramidal tract within 14 days following stroke. In conclusion, very fine anatomical structures are visible on 3DAC images, and it can be used as a diagnostic tool for the human brain. PMID- 9745852 TI - Clinical features of polymyositis/dermatomyositis with steroid-resistant interstitial lung disease. AB - Polymyositis and dermatomyositis (PM/DM) without creatine kinase (CK) elevation shows a poor prognosis. PM/DM is complicated with interstitial lung disease (ILD), some of which progress rapidly. To clarify the clinical features of PM/DM from the viewpoint of ILD progression, the clinical data of 25 PM/DM patients with ILD were reviewed. They were classified as responders or non-responders. The patients whose ILD responded to steroid therapy and elicited good clinical courses were termed as responders. On the other hand, the patients who had rapidly progressive ILD resistant to steroid therapy were considered as non responders. The patients diagnosed to have DM were likely to be steroid resistant. The non-responder group revealed significantly high aspartate aminotransferase (AST), low CK, low white blood cell (WBC), and low absolute lymphocyte counts in their peripheral blood. High CK/AST may be a favorable predictor of the disease. The percentages of lymphocytes in bronchoalveolar lavage fluid were increased in both groups. However, the percentages of two responders with low CK/AST were lower than those of three non-responders. A steroid-resistant ILD group with PM/DM may be clinically different from a steroid responsive ILD group. PMID- 9745853 TI - Isolated partial growth hormone deficient short stature with syringomyelia not associated with birth injury. AB - A 59-year-old female with 20-year history of slowly progressing muscle atrophy and sensory disturbance of upper extremities showed short stature, scoliosis, hunger type of sensory dissociation of the upper extremities and pyramidal tract sign of the lower extremities. Magnetic resonance imaging (MRI) clarified hypoplasia of the anterior pituitary lobe, Arnold-Chiari malformation and cervical syringomyelia. Insulin and arginine stimulating tests revealed partial type of isolated growth hormone (GH) deficiency but GH gene analysis detected no defects of GH genes. It was considered to be a rare case of non-hereditary hypopituitarism with Chiari malformation and syringomyelia not associated with perinatal injury, namely a midline anomaly syndrome. PMID- 9745854 TI - Unusual accumulation of glycogen in liver parenchymal cells in a patient with anorexia nervosa. AB - Anorexia nervosa is an eating disorder characterized by a fear of weight gain and a preoccupation with body image. Although hepatic involvement has been reported in patients with anorexia nervosa, the mechanism is not fully understood. We describe a patient with anorexia nervosa with liver function abnormalities. Light and electron microscopic observations revealed a remarkable accumulation of glycogen in hepatocytes. These results suggest that adaptive responses to starvation may alter carbohydrate metabolism in patients with anorexia nervosa. PMID- 9745855 TI - Central diabetes insipidus associated with a missense mutation in the arginine vasopressin gene that replaces Ala at the carboxyterminus of the signal peptide with Thr. AB - We report an 18-year-old male with a history of polyuria, polydipsia, and thirst since childhood. In a hypertonic saline infusion test, the patient's plasma vasopressin rose only to 0.28 pg/ml. In a water deprivation test, his urinary osmolality rose only to 189 mosmol/kg and then rose to 538 mosmol/kg by vasopressin administration. A T1-weighted magnetic resonance imaging (MRI) scan revealed a loss of the posterior pituitary bright spot. Sequencing of the vasopressin gene showed a heterozygous point mutation that replaced Ala at the carboxyterminus of the signal peptide with Thr. His father also had similar history, and we therefore diagnosed his illness as familial central diabetes insipidus. PMID- 9745856 TI - Three cases of primary pulmonary amyloidosis. AB - In three cases of primary pulmonary amyloidosis the chief complaint was hemosputum. The diagnosis of amyloidosis was made using histochemical analysis of bronchial wall biopsy in all cases; multiple nodular lesions were observed in trachea and bronchi on flexible fiberoptic bronchoscopy. The surface of the tracheobronchial mucosa was smooth but bled easily. In one patient, chest X-ray film showed a solitary nodular shadow in the left lower lung field. These three cases were tracheobronchial amyloidosis, and one case was combined with nodular parenchymal type amyloidosis. PMID- 9745857 TI - Retroperitoneal Castleman's disease of the hyaline vascular type presenting arborizing calcification. AB - Castleman's disease (CD) usually manifests as a solitary mediastinal tumor and only rarely as an isolated retroperitoneal mass. In the latter instances it is difficult to distinguish radiographically from other retroperitoneal masses. We report a 22-year-old female patient with retroperitoneal CD of the hyaline vascular type presenting with arborizing calcification. This characteristic calcification pattern is considered unique to CD, and is useful in diagnosis when present. PMID- 9745858 TI - Hepatic veno-occlusive disease in a case of polymyositis associated with thrombotic thrombocytopenic purpura/hemolytic uremic syndrome. AB - A 50-year-old woman was treated with prednisolone for polymyositis. During the therapy, thrombotic thrombocytopenic purpura/hemolytic uremic syndrome (TTP/HUS) occurred. Neither plasma infusion nor plasma exchange could relieve the clinical manifestations of TTP/HUS. Moreover, massive ascites appeared and worsened her condition. She died approximately one year after the diagnosis of polymyositis. The autopsy revealed centri-lobular hepatic necrosis and nonthrombotic obliteration of hepatic small veins. The diagnosis of hepatic veno-occlusive disease (VOD) was made. It was suspected that common factors other than cytoreductive therapy had damaged the endothelium and caused TTP/HUS and VOD in a case of polymyositis. PMID- 9745859 TI - Fatal hyperammonemia in a patient with systemic lupus erythematosus. AB - We treated a 31-year-old woman with systemic lupus erythematosus, renal failure with nephrotic syndrome, and a long-standing seizure disorder, who developed severe hyperammonemia with a fatal outcome. Blood chemistry examination did not indicate liver disease, and amino acid concentrations did not suggest a defect in the urea cycle. Discontinuation of anticonvulsant treatment with valproic acid (VPA) failed to bring about improvement. We speculated that hyperammonemia in this case was induced by VPA, and the existence of other underlying factors, including the administration of aspirin and cimetidine, hypoalbuminemia, and renal failure might elevate the concentration of the serum free fraction of VPA. PMID- 9745860 TI - Primary antiphospholipid syndrome with recurrent transient ischemic attacks: report of a case and its successful treatment. AB - A 35-year-old woman was admitted to our hospital with complaints of a two-year history of recurrent, daily episodes of transient ischemic attacks; the symptoms consisted of scotoma of her left eye, vertical diplopia, and paresthesia of her right arm. The presence of lupus anticoagulants and anticardiolipin antibodies led to the diagnosis of antiphospholipid syndrome (APS). After thrombotest values had decreased to 30% (international normalized ratio: 1.5) with warfarin, her symptoms did not recur. This suggests that anticoagulant therapy is effective for the prevention of recurrence of ischemic events complicated by primary APS, even when they occur repeatedly. PMID- 9745861 TI - Human parvovirus B19 infection resembling systemic lupus erythematosus. AB - A 39-year-old female visited our hospital because of morning stiffness, arthralgia, skin rash of the extremities and general fatigue. On examination, she also had a malar rash and an oral ulcer. Laboratory findings revealed that antinuclear antibodies were positive and complement component levels (C3, C4, CH50) were all low. Serology for human parvovirus B19 (HPV-B19) was positive for both immunoglobulin (Ig)M and IgG. Gradually, her symptoms improved and laboratory data returned to normal range without medications. This case suggests that HPV-B19 infection may be attributed to the pathogenesis of systemic lupus erythematosus. PMID- 9745862 TI - An autopsy case of Wegener's granulomatosis with pachymeningitis. AB - A 61-year-old woman presented with high fever, headache and left facial palsy with diplopia. Histopathological examination of the biopsied specimens taken from nasal mucosa and kidney revealed a granulomatous angiitis with giant cell infiltration. Ga-DTPA-enhanced magnetic resonance imaging (MRI) revealed a thickening of dura mater in the middle cranial fossa and tentorium cerebelli. The observed left facial and occulomotor palsy was considered to be caused by pachymeningitis associated with Wegener's granulomatosis (WG). Cyclophosphamide combined with prednisolone effectively improved the symptoms. However, the patient died of acute interstitial pneumonitis, presumably caused by cyclophosphamide. The pathohistology obtained in the autopsy revealed a fibrous thickening of the dura mater in the left meningen with a segmental scarring of the arteries and a necrotizing arteritis in the kidney. PMID- 9745863 TI - Extrahepatic portal-systemic encephalopathy without portal hypertension. PMID- 9745864 TI - Effects of growth hormone in short children after renal transplantation. French Society of Pediatric Nephrology. AB - From 1991 to 1993, 90 children having received a kidney graft with a post transplantation period of at least 12 months were included in a prospective study carried out in 18 French pediatric centers. After informed consent and randomization, children received recombinant human growth hormone (rhGH) (Genotonorm, Pharmacia peptide hormones) 30 U/m2 per week, either immediately on enrollment, for the treated group, or after 1 year of follow-up for the group serving as a control. After 1 year both groups were treated and we analyzed data during the subsequent years. Eighty-five children completed the 1-year study. Growth velocity was significantly increased by rhGH: 7.7 cm with a gain of +0.3 standard deviation score in the treated group versus 4.6 cm in the control group (P<0.0001) during the 1st year. Four factors predicted response to therapy: growth velocity prior to GH therapy, glomerular filtration rate (GFR) at the start, mode of corticosteroid administration, and degree of insulin resistance. After 1 year we observed a moderate, significant decrease in GFR in both groups. Biopsy-proven acute rejection episodes were not significantly more frequent during the 1st year in the group of patients who received rhGH: 9 in 44 versus 4 in 46 patients. The patients who rejected did not differ in terms of age, renal function at the start, and type of immunosuppression, but history of rejection before GH treatment was discriminatory: 6 of 17 children with two or more episodes had a new rejection versus 1 of 22 who had no or only one episode (P=0.01). Glucose tolerance was not modified after 1 year of GH therapy. During the subsequent years of treatment a decrease in growth velocity was noted: 5.9 cm at 2 years, 5.5 at 3 years, and 5.2 cm at 4 years. In conclusion, GH is efficient for improving growth velocity in short transplanted children, inducing clear-cut but limited catch-up growth. The risk of rejection was shown only in patients with a prior history of more than one rejection episode. PMID- 9745865 TI - Sporting activity following kidney transplantation. AB - A postal questionnaire on sporting activity following renal transplantation revealed that most units encouraged participation in the majority of sports, but counselled against contact sports such as rugby football, boxing, and Asian martial arts. PMID- 9745866 TI - In utero nephropathy, Denys-Drash syndrome and Potter phenotype. AB - We report an unusual case of Denys-Drash syndrome presenting in a newborn infant with end-stage renal failure of antenatal origin and Potter phenotype. DNA analysis showed a novel missense change in arginine 394 of zinc finger 3 of the WT1 gene. This mutation may lead to an earlier and more severe presentation of Denys-Drash syndrome. It may be of interest to look for this mutation in other Potter phenotype cases. PMID- 9745867 TI - Oxygen-dependent injury by a human plasma factor associated with minimal change disease. AB - The mechanism by which a human plasma factor associated with proteinuria is able to cause experimental glomerular albumin leakage is unknown. This factor (called 100KF) is able to induce glomerular alterations in the rat kidney, similar to those seen in minimal change disease, including loss of glomerular sialoglycoproteins and decreased expression of glomerular ecto-ATPase. It was previously shown that 100KF is able to stimulate release of reactive oxygen species in inflammatory cells in vitro. This prompted us to test whether 100KF induced injury is oxygen dependent. The expression of glomerular sialoglycoproteins and ecto-ATPase was evaluated by standard histochemistry and computerized image analysis and expressed in arbitrary units. Rat kidney sections were incubated with or without 100KF under normal or oxygen-poor, i.e., nitrogen, conditions, or with supplementation of superoxide dismutase (SOD, 100 U/ml). The effect of 100KF on glomerular ecto-ATPase was oxygen dependent (32.98+/-2.14 under air vs. 65.20+/-5.53 under nitrogen, P< or =0.01), in contrast to the 100KF induced loss of glomerular sialoglycoproteins that was not significantly altered under nitrogen (62.67+/-10.08 under air vs. 61.74+/-26.05 under nitrogen). Supplementation of SOD to 100KF solution under normal incubation conditions also suggested oxygen-dependent impairment of glomerular ecto-ATPase. Alternate perfusion ex vivo of the rat kidney with 100KF followed by diluted plasma showed that enhanced leakage of plasma proteins could be inhibited with SOD, indicating oxygen dependency of this 100KF-induced enhanced permeability (60.25+/-19.32 microg urinary albumin/ml after 100KF perfusion vs. 25.23+/-12.05 microg/ml after 100KF plus SOD, P< or =0.01). We conclude that the action of 100KF upon specific glomerular matrix molecules is oxygen dependent, as is the albumin leakage induced by 100KF in the present ex vivo model. PMID- 9745868 TI - Levamisole treatment in steroid-sensitive and steroid-resistant nephrotic syndrome. AB - Since 1992 we have treated 11 children with frequently relapsing steroid sensitive (n=6) or steroid-resistant (n=5) nephrotic syndrome with levamisole. All had been non-responsive to other immunosuppressive medication before levamisole treatment. All steroid-sensitive patients had signs of steroid toxicity. At least 1 kidney biopsy had been performed prior to study in each patient. Five children had minimal glomerular changes and the other 6 focal segmental glomerular sclerosis. The patients were treated with levamisole (2.5 mg/kg per 48 h) for at least 2 months (up to 18 months, median 10 months). Two patients had additional immunosuppression (cyclosporine A) during levamisole treatment. All patients with steroid-sensitive nephrotic syndrome became free of proteinuria within 2 months and have remained in remission after discontinuation of levamisole (follow-up time 8-50 months, median 24 months). None of the children with steroid-resistant nephrotic syndrome experienced a remission. Side effects were observed in 2 patients and included a granulocytopenia and a severe psoriasis-like cutaneous reaction; both were reversible after discontinuation of levamisole. We conclude that levamisole is of benefit in steroid-sensitive nephrotic syndrome but not in steroid-resistant nephrotic syndrome. PMID- 9745869 TI - Interleukin-12 and interferon-gamma production in childhood idiopathic nephrotic syndrome. AB - Cellular immune disturbances, and T lymphocyte function in particular, have been previously implicated in idiopathic nephrotic syndrome (INS) of childhood. There are different patterns of cytokine expression in various forms of glomerulonephritis, which suggests that local production of these peptides plays an important role in the pathogenesis and progression of glomerulonephritis. To investigate T-cell and monocyte/macrophage cytokine production in INS, interleukin-12 (IL-12) and interferon-gamma (IFN-gamma) production by peripheral blood mononuclear cells (PBMC) of 11 children with steroid-sensitive nephrotic syndrome (SSNS), 9 with focal segmental glomerulosclerosis (FSGS), and 17 healthy controls was determined. Children with SSNS were studied in relapse, during corticosteroid treatment, and in stable remission, off corticosteroid treatment. IL-12 was not detected in serum, urine, and in supernatants of unstimulated PBMC. IL-12 production by concanavalin A (Con A)-stimulated PBMC of children with SSNS and FSGS was not different from controls. IFN-gamma production by Con A stimulated PBMC was decreased in children with relapsing SSNS, both in relapse and and during corticosteroid treatment. However, in stable remission it was similar to controls. Markedly decreased IFN-gamma production (P<0.001) was observed by pokeweed mitogen-stimulated PBMC of relapsing SSNS patients and moderately decreased production by PBMC of FSGS patients. This study has established a decreased production of IFN-gamma by PBMC of relapsing SSNS and FSGS patients, but does not allow differentiation between these two different conditions. IL-12 did not have a pathogenic role in either SSNS or FSGS. PMID- 9745870 TI - Influence of age at onset on the outcome of steroid-sensitive nephrotic syndrome. AB - To investigate whether age at onset of steroid-sensitive nephrotic syndrome (SSNS) is predictive of subsequent relapses, or influences outcome, we retrospectively studied 60 patients who were under 10 years of age at onset and were followed for over 10 years. They were divided into three groups according to age at diagnosis: group 1-3 (1.0-3.9 years at onset, n=24), group 4-6 (4.0-6.9 years at onset, n=22), and group 7-9 (7.0-9.9 years at onset, n=14). In the 51 patients with long-term remission, defined as remaining relapse-free over 3 years, the total number of relapses was significantly more in group 1-3 (n=18) than in group 4-6 (n=19), and the interval between onset and long-term remission was significantly longer. Group 4-6 and group 7-9 had fewer patients with active disease at 10 years, follow-up than group 1-3, as assessed by the Kaplan-Meier method. These data suggest that the age at onset of SSNS influences the clinical course (i.e., frequency of relapses) and the time to reach long-term remission. An age of less than 4 years at onset of SSNS is associated with greater likelihood for frequent relapses and a greater time interval to attain long-term remission. PMID- 9745871 TI - Glomerular deposits and hypoalbuminemia in acute post-streptococcal glomerulonephritis. AB - In a search for correlations between glomerular morphology and clinical manifestations in acute post-streptococcal glomerulonephritis, data for 40 biopsied patients were reviewed. A major correlation was observed between severe hypoalbuminemia and the absence of deposits on the paramesangial portion of the glomerular basement membrane. Subepithelial deposits were present on both the capillary loop and paramesangial segments of the basement membrane in 29 patients, whereas in 11 the deposits were present only on the capillary loop. Patients with paramesangial segments devoid of deposits had a mean (+/-1 SD) nadir serum albumin level of 1.90 (+/-0.40) g/dl, whereas the mean nadir level in those with deposits in both locations was 2.83 (+/-0.64) g/dl (P<0.001). Also significant was the difference in paramesangial deposit scores in patients with nadir serum albumin levels < or =2.5 g/dl versus those with levels >2.5 g/dl. The amount of subepithelial deposit on the capillary loop was not significantly different in the two groups, and no correlation was found between loop deposits and serum albumin levels. Except for greater edema and significantly less frequent gross hematuria in those with absent paramesangial deposits, clinical and laboratory findings for the two groups did not differ. PMID- 9745872 TI - Prevalence of microalbuminuria in children with sickle cell disease. AB - Renal involvement is common in homozygous sickle cell disease (HbSS), including glomerular hypertension and hypertrophy similar to that seen in rodent models of ablative nephrectomy and stage I diabetic nephropathy (DN). The proteinuria in the rodent models is attenuated by angiotensin converting enzyme inhibition (ACEI). Microalbuminuria (MA) is a sensitive marker for renal involvement in DN prior to the development of proteinuria, and is also attenuated with ACEI. Elevated urinary microalbumin/creatinine ratios (U Alb/Cr) >20 mg/g Cr are reported in 39%-43% of adults with HbSS, and studies are ongoing in this age group to assess the effect of attenuated proteinuria by ACEI on long-term renal function. The purpose of this study was to prospectively investigate the prevalence of MA in children with HbSS and determine factors which affect its expression. U Alb/Cr values were measured on spot urine samples in 102 children (aged 2-18 years, mean 9.47+/-4.62, M:F=53:49) by rate nephelometry. Children with prior known proteinuria, hypertension, or fever/pain episode in the last 15 days were excluded. MA was present in 26.5% of all children with HbSS. However, in children between the ages of 10 and 18 years, the prevalence was 46% (similar to the prevalence in adults). There was a strong correlation between patient age and prevalence of MA (P<0.0001) by both univariate and multivariate analysis. However, pain frequency, hospitalization, transfusion program, ferritin levels, and Cr clearance (C(Cr)) did not correlate with prevalence, although C(Cr) (as estimated by Schwartz formula) was elevated in all. We conclude that the prevalence of MA in the 2nd decade of life is similar to that in adults. PMID- 9745873 TI - Hepatitis B virus-associated nephropathy in black South African children. AB - Hepatitis B virus (HBV)-related membranous nephropathy (MN) is prominent in secondary nephrotic syndromes (NS) in Africa, but the features of this disease and the spectrum of associated glomerulopathies have not been adequately documented in black children. HBV was detected in 93 children with NS included in this study. In 70 patients the histological type was membranous; 46 were followed for a mean of 3.4 years (range 1-11 years). Spontaneous elimination of both HBsAg and HBeAg occurred in 10 (21.7%) patients; 16 (34.8%) cleared HBeAg alone. Co existing liver disease occurred in 18 (25.7%) and hypocomplementemia (C3,C4) in 47.1% and 11.4% of children, respectively. Sixty-five (92.9%) patients had normal renal function, 1 (1.4%) impaired renal function, 3 (4.3%) chronic renal insufficiency, and 1 (1.4%) end-stage renal disease at last hospital visit. Twelve patients were in remission, all having cleared HBeAg. HBVMN was clinically indistinguishable from 24 children with idiopathic MN, although biochemical characteristics were different. This report delineates the natural history of HBV infection in black South African children with NS, the majority of whom have MN. Disease remission in HBVMN parallels elimination of HBV antigens, particularly HBeAg. Comparison of HBVMN with idiopathic MN revealed clinically indistinguishable characteristics but unexplained biochemical differences. PMID- 9745874 TI - Renal function in Inuit survivors of epidemic hemolytic-uremic syndrome. AB - We undertook a case-control study to evaluate the renal health of survivors of hemolytic-uremic syndrome (HUS) from the 1991 Arctic epidemic of Escherichia coli O157:H7 gastroenteritis 4 years after the epidemic. Eighteen children who developed HUS during the 1991 epidemic and 18 age- and sex-matched controls from the same community who had uncomplicated gastroenteritis were compared in 1995 for height, weight, blood pressure, urinalysis, and glomerular filtration rate (GFR), measured using continuous subcutaneous infusion of non-radioactive iothalamate. HUS survivors did not differ from controls in height, weight, systolic (HUS 118 mmHg, control 117 mmHg) or diastolic (HUS 64 mmHg, control 62 mmHg) blood pressures. Hematuria was detected more frequently in HUS survivors (11/18 vs. 4/18, P<0.05), but no child had proteinuria. Mean GFR did not differ between the two groups (HUS 159 ml/min per 1.73 m2, control 147 ml/min per 1.73 m2). Survivors of post-enteritic HUS from the 1991 Arctic E. coli 0157:H7 outbreak have excellent renal function 4 years after the epidemic. PMID- 9745875 TI - Pitfalls in measuring inulin and para-amino-hippuric acid clearances. AB - Various carbohydrates and a variety of widely used medicines interfere with the generally used laboratory methods for determining inulin and para-aminohippuric acid (PAH). When these pitfalls are not recognized, false measurements of inulin and PAH clearances, which represent glomerular filtration rate and renal plasma flow respectively, are obtained. When performing these tests a careful history of dietary habits and oral drug therapy must be taken. PMID- 9745876 TI - The Fanconi syndrome of cystinosis: insights into the pathophysiology. AB - Cystinosis is a lysosomal storage disease which is the most-common inherited cause of the Fanconi syndrome. Insights into the pathophysiology of the proximal tubular defect have come from in vitro studies of the cystine-loaded tubule. Proximal tubules loaded with cystine have a generalized proximal tubule transport defect characteristic of the Fanconi syndrome. The decrease in proximal tubular transport with cystine loading is not due to an increase in paracellular permeability with backflux of solute transport from the blood to the tubular lumen, but due to a decrease in active transport. The Na-K-ATPase activity is intact under Vmax conditions in cystine-loaded tubules; however, the production of ATP is severely compromised. The cystine-loaded tubule has a lower intracellular phosphate concentration than that of control tubules. This low intracellular phosphate concentration in cystine-loaded tubules likely plays a critical role in the decrease in intracellular ATP. Preservation of intracellular phosphate at control levels prevents the decrease in intracellular ATP and the proximal tubule respiratory dysfunction with cystine loading. The clinical significance and future directions for investigation are discussed. PMID- 9745877 TI - Current concepts and controversies in IgA nephropathy. PMID- 9745878 TI - Nitric oxide: from molecular biology to clinical nephrology. PMID- 9745879 TI - Membranoproliferative glomerulonephritis type III. PMID- 9745880 TI - Arterial hypertension with normal urinalysis in Henoch-Schonlein disease: a further case. PMID- 9745881 TI - Atrial flutter? An electrocardiographic artifact caused by the COBE PRISMA system. PMID- 9745882 TI - Successful surgical treatment of sclerosing peritonitis persisting after renal transplantation. PMID- 9745883 TI - The natural history of HIV disease in haemophilia. AB - Patients with haemophilia, particularly that due to factor VIII deficiency, have been exposed to a wide range of infective agents transmitted through blood products that have in other ways revolutionized their care. The most devastating of these transfusion transmitted infections has been the human immunodeficiency virus (HIV). AIDS in haemophilic patients was first described in 1982 and it has significantly reduced the life expectancy of these patients. In this article, the impact of the HIV epidemic within haemophilic patients treated with coagulation factor concentrate is discussed. The effect of age at time of exposure to HIV and the value of disease markers such as P24 antigenaemia and CD4 counts are considered in detail. The relationship between HIV disease and coexisting hepatitis C infection is described and the incidence of secondary malignancies such as lymphoma is reviewed. In this patient population the recent elucidation of the life cycle and dynamics of HIV as well as the technological advances in the development of the HIV RNA PCR assay for HIV viral load have revolutionalized the diagnosis, prognosis, management and treatment of HIV infection. PMID- 9745884 TI - Acute myeloid leukemia in the elderly. AB - The optimal management of acute myeloid leukemia in patients over 60 years-of-age remains a controversial issue. The complete remission rates after conventional induction chemotherapy progressively decreases after the age of 60. This is explained by host-related factors and by differences in the biology of leukemia. The incidence of adverse prognostic factors (trilineage myelodysplasia, unfavorable karyotype, mdrl-positive phenotype) is higher in elderly patients. Three strategies are currently offered to older adults with acute myeloid leukemia: intensive chemotherapy, palliative treatment and attenuated dose chemotherapy. Currently, complete remission rates achieved with conventional chemotherapy range from 40-65% according to inclusion criteria. In the past few years, two approaches have been tested in order to improve the results of induction chemotherapy: modifications of chemotherapy regimens with new intercalating agents (idarubicin, mitoxantrone) and the use of myeloid growth factors. Myeloid growth factors have been administered with two objectives: to reduce the duration of neutropenia and the toxic death rate when given after induction chemotherapy, and to prime leukemic blasts when given during chemotherapy. The results of published placebo-controlled studies are discussed. The issues of palliative treatment and of attenuated dose chemotherapy are also addressed in the review, with special emphasis on the role of oral idarubicin. PMID- 9745885 TI - Lymphomas and reactive lymphoid lesions in HIV infection. AB - Infection by the human immunodeficiency virus (HIV) causes depletion of CD4 positive lymphocytes with consequent immunodeficiency. HIV infection also causes, by direct or indirect mechanisms, both reactive and neoplastic changes in lymphoid tissues. In primary infection reactive changes are a direct response to HIV. Later in the course of the disease there are reactive changes in lymph nodes and extranodal lymphoid tissues which are likely to be largely an indirect effect of HIV infection, being a response to opportunistic infection by other organisms. There is also an increased incidence of autoimmune phenomena in HIV-infected subjects which is likely to be consequent, at least in part, on impaired control of the proliferation of self-reactive B-cell clones. A second mechanism of immune damage of blood cells, probably operating in the case of HIV-related immune thrombocytopenic purpura, is that of cellular damage by immune complexes containing antiviral antibodies. Lymphoid neoplasms associated with HIV infection include non-Hodgkin's lymphoma, Hodgkin's disease and, uncommonly, plasma cell dyscrasias. HIV-associated lymphomas have distinct clinicopathological features and generally a poor prognosis. As for reactive lymphoid lesions, induction of neoplasia is likely, in the majority of cases, to be an indirect rather than a direct effect of the virus. The combination of chronic B-cell stimulation and impaired T-cell function is important, and interaction of lymphoid cells with virus-infected stromal cells may also play a role. Infection by oncogenic viruses such as the Epstein-Barr virus and human herpes virus 8 is also aetiologically important. In rare cases of T-cell lymphoma, HIV may be directly oncogenic. PMID- 9745886 TI - Severe thalassaemia intermedia: clinical problems in the absence of hypertransfusion. AB - In many of the parts of the world where thalassaemia is common, the blood supply is inadequate or unsafe, and desferrioxamine is too expensive for routine use. We classify some patients as having 'severe thalassaemia intermedia', i.e. those with moderately severe thalassaemia who can survive without regular transfusions, but who are at risk of many complications which are reviewed here. These include bone deformity and fractures, extramedullary haemopoietic tumours, leg ulcers, autoimmune haemolysis and, especially after splenectomy, thromboembolism and infection. An increase in the quality and safety of the blood supply, and a cheaper and/or oral iron chelator, would enable more of these patients to be treated as thalassaemia major and have improved survival and quality of life. PMID- 9745887 TI - Transfusion virology: progress and challenges. AB - Discoveries of new human viruses and new technologies for their detection have made, and will continue to make, major contributions to the safety of blood transfusion. This article discusses the practical issues involved in the implementation of additional serological screening tests for viruses such as human T-lymphotropic virus, and reviews current information on the prevalence and pathogenicity of more recently discovered viruses, such as hepatitis G virus (HGV) or GB virus-C (GBV-C) and human herpes virus 8, a potential aetiological agent of Kaposi's sarcoma. Progress in the technology behind nucleic acid amplification techniques, such as the polymerase chain reaction (PCR), makes direct detection of viruses such as human immunodeficiency virus and hepatitis C virus possible. The use of such methods for screening will allow the earlier detection of acutely-infected individuals and the elimination of transmission from 'window' period donations before seroconversion for antibody. Establishing a framework for PCR-based screening would also enable the testing for others such as hepatitis A virus, parvovirus B19 and GBV-C/HGV for which serological detection methods cannot be or have not been developed. PMID- 9745888 TI - Congenital dyserythropoietic anaemias: clinical features, haematological morphology and new biochemical data. AB - Three types of congenital dyserythropoietic anaemia (CDA) were originally identified on the basis of the pattern of dysplastic changes in the erythroblasts and the results of the acidified serum lysis test (Ham test). These were designated CDA types I, II and III. Several other types have been described subsequently and new forms continue to be reported. Some patients with CDA develop iron overload even without repeated blood transfusion and may present with the complications of severe iron overload. Dysmorphic features are seen in some cases, especially of CDA type I. In CDA type II, incomplete processing of N linked oligosaccharides leads to a marked reduction of polylactosamines associated with band 3 of the red cell membrane. A few cases of CDA type III develop lymphoid neoplasms. Some of the Swedish cases of CDA type III have developed a retinal abnormality characterized by angioid streaks and macular degeneration. The chromosomal localizations of the disease gene in CDA types I and II and in the Swedish family with CDA type III are now known, but the identities of the mutant genes are still unknown. Cases of CDA type I have shown a partial haematological response to interferon-alpha, however the biochemical basis of this response is unclear. An important step in the diagnosis of sporadic cases of CDA is the exclusion of known causes of acquired dyserythropoiesis. PMID- 9745889 TI - Ion-RNA interactions in the RNA pseudoknot of a ribosomal frameshifting site: molecular modeling studies. AB - The three-dimensional (3-D) structure of a RNA pseudoknot that causes the efficient ribosomal frameshifting in the gag-pro region of mouse mammary tumor virus (MMTV) has been determined recently by nuclear magnetic resonance (NMR) studies. But since the structure refinement in the studies did not use metal ions and waters, it is not clear how metal ions participate in the stabilization of the pseudoknot, and what kind of ion-RNA interactions dominate in the tertiary contacts for the RNA pseudoknotting. Based on the reported structure data of the pseudoknot VPK of MMTV, we gradually refined the structure by restrained molecular dynamics (MD) using NMR distance restraints. Restrained MD simulation of the RNA pseudoknot was performed with sodium ions and water molecules. Our results are in good agreement with known NMR data and delineate the importance of the metal ion coordination in the stability of the pseudoknot. In the non coaxially stacking pseudoknot, stem 1 (S1), stem 2 (S2), and the intervening A14 involves unconventional stacking of base pairs coordinated by Na+ and/or bridging water molecules. A6 and G7 of loop L1 make a perfect base stacking in the major groove and are further stabilized by coordinated Na+ ions and water molecules. The first 4-nucleotide (nt) ACUC of loop L2 form a sharp turn and the following 4 nt AAAA cross the minor groove of S1 and are steadied by interactions with the nucleotides of S , bridging water molecules and coordinated Na+ ions. Our studies suggest that the metal ion plays a crucial role in the RNA pseudoknotting of VPK. In the stacking interior of S1 and S2, the Na+ ion is positioned in the major groove and interacts directly with the carbonyl group O6 of G28 and carbonyl group O4 of U13 in the wobble base pair U13:G28. The ion-RNA interactions in MMTV VPK not only stabilize the RNA pseudoknot but also modify the electrostatic properties of the nucleotides at the critical parts of the pseudoknot VPK. PMID- 9745890 TI - Probing site-specific interactions in protein-DNA complexes using heteronuclear NMR spectroscopy and molecular modeling: binding of Cro repressor to OR3. AB - In this paper, a general method is developed to study site-specific interactions in DNA-protein complexes using heteronuclear NMR spectroscopy and molecular modeling. This method involves two steps: (a) homonuclear 1H NMR and molecular modeling are used to develop a low resolution model and (b) 15N7-guanosine containing oligonucleotides are employed to probe the specific intermolecular interactions predicted in (a). This method is applied to Cro-operator complex due to its small size and extensive prior characterization. Non-exchangeable and exchangeable base protons have been assigned by nuclear Overhauser effect spectroscopy (NOESY) and chemical shift correlation spectroscopy. Extensive line broadening has been observed in the 1H NMR spectra of the operator DNA in the presence of protein. Differential line-broadening observed in the imino proton region and the comparison of NOESY spectra in the presence and absence of Cro protein show that guanosine-12 and guanosine-14 are involved in the Cro-DNA interaction, while the three A.T base-pairs at the 3'- and 5'-termini play only a minor role in the binding. A model of the Cro-operator DNA complex has been constructed by docking helix-3 of the Cro protein in the major groove and it predicted specific hydrogen bonds between N7 of guanosines-12 and -14 and the side-chain of Lys-32 and Ser-28, respectively. The appearance of a new resonance in the temperature dependent proton detected heteronuclear multiple quantum coherence (HMQC) spectra of the Cro-DNA complex also demonstrates a specific interaction of Cro with guanosine-14 of the operator DNA. PMID- 9745891 TI - 23Na NMR study of the interaction between DNA and the platinum (II) compounds: cis-DDP, trans-DDP and TDP. AB - The interaction of calf thymus DNA with cis-DDP, trans-DDP and TDP was studied by 23Na NMR in aqueous solutions at pH=7.0, with Pt(II) compounds/DNA(P) (P=Phosphate) molar ratios r increasing from 0 till to 1P. 23Na NMR results are interpreted on one hand, with the help of a " two states model " with R(F) and R(B) relaxation rates, and, on the other hand, using the " entropy of fluctuations " concept developed by Lenk. We have established that, for the studied platinum compounds, the preference to interact with DNA phosphate sites interpreted as a perturbation of the counterions environment- is in a decreasing order: TDP >> cis-DDP > trans-DDP. These results are discussed with regard to the interaction of DNA with the hard bication Mg++ and the soft bication Cu++. PMID- 9745892 TI - Localization of histone H1 binding sites within the nucleosome by UV-induced H1 DNA crosslinking in vivo. AB - In our previous paper (Belikov et al., (1993), Nucl. Acids Res., 21, 4796-4802) we had studied DNA-protein architecture of so-called Alu-repeats in D. melanogaster ribosomal nontranscribed spacer using DNase I genomic footprinting and UV-induced DNA-protein crosslinking. Our data indicated precise positioning of two non-histone proteins (rABP50 and rABP 70), histone octamer, and histone H1 within the sequences of Alu-repeats. The data on the histone H1 binding sites within Alu-repeat region was presented, but not discussed as the authors could not reach a consensus in its evaluation. Here, the authors use these data to present a model of placement of the linker histone(s) within nucleosome. Our model places one contact of the globular domain of linker histone in the major groove on the inside of DNA superhelix just within the end of the core particle (site +6.5) and the second, close to the dyad (site -1). C-terminal tail binds to the internucleosomal spacer and N-terminal tail interacts simultaneously with the adjacent gyres thus stabilizing DNA superhelix. PMID- 9745893 TI - A theoretical perusal of the satellite DNA curvature in tenebrionid beetles. AB - The curvature patterns of seven satellite DNAs taken from beetles belonging to the family Tenebrionidae (Coleoptera) were modelled utilising a number of computer programs that describe and plot the curvature profiles of DNA. The theoretical analysis agreed with the experimentally observed curvature of most of these satellite DNAs, and its absence in Tribolium freemani and Tenebrio obscurus satellite I. In many cases, the tenebrionid satellite DNAs lack periodically repeated runs of phased-A-tracts, yet they represent a clear example of curved DNA. The macroscopic curvature of satellites from these closely related organisms confirmed that other sequence elements must be participating in the bending of these DNAs. Our modelling approaches are discussed, together with previous experimental results, in terms of the role played by DNA curvature in the organisation of satellite DNA and the tight compacting of heterochromatin. PMID- 9745894 TI - The base contents of A, C, G or U for the three codon positions and the total coding sequences show positive correlation. AB - The distribution of the occurrence frequencies of each of the four bases at the first, second and third codon positions and in the total coding sequences is analyzed by a graphic method. It is shown that for the coding sequences of 90 species, A has its largest frequency at the second codon position and the smallest one at the third position. C and U have their least frequencies at the first codon position, while G has its largest frequency at the first codon position. By this method, we also find that for each base, there is positive correlation between every two frequencies of the base in the first, second, third and the total coding sequences for 90 species. For each of the four bases, the correlation between the frequencies at the third codon position and that in the total coding sequences is more prominent than others. A statistical method is used to give a precise description of the correlation for the frequencies of every base and it is found that the conclusions drawn by the graphic method are consistent with that got by the statistical method. PMID- 9745895 TI - Melting of cross-linked DNA. III. Calculation of differential melting curves. AB - In our previous papers I and II (D. Y. Lando et al, J. Biomol. Struct. Dynam. (1997) v. 15, N1, p. 129-140, p. 141-150), two methods were developed for calculation of melting curves of cross-linked DNA. One of them is based on Poland's and another on the Fixman-Freire approach. In the present communication, III, a new theoretical method is developed for computation of differential melting curves of DNAs cross-linked by anticancer drugs and their inactive analogs. As Poland's approach, the method allows study of the influence of the loop entropy factor, delta(n), on melting behavior (n is the length of a loop in base pairs). However the method is much faster and requires computer time that inherent for the most rapid Fixman-Freire calculation approach. In contrast to the computation procedures described before in communications I and II, the method is suitable for computation of differential melting curves in the case of long DNA chains, arbitrary loop entropy factors of melted regions and arbitrary degree of cross-linking including very low values that occur in vivo after administration of antitumor drugs. The method is also appropriate for DNAs without cross-links. The results of calculation demonstrate that even very low degree of cross-linking alters the DNA differential melting curve. Cross-linking also markedly strengthens the influence of particular function delta(n) upon melting behavior. PMID- 9745896 TI - Molecular structure of two crystal forms of cyclic triadenylic acid at 1A resolution. AB - The three dimensional structures of cyclic deoxytriadenylic acid, c-d(ApApAp), from two different trigonal crystal forms (space groups P3 and R32) have been determined by x-ray diffraction analysis at 1A resolution. Both structures were solved by direct methods and refined by anisotropic least squares refinement to R factors of 0.109 and 0.137 for the P3 and R32 forms, respectively. In both crystal forms, each of the two independent c-d(ApApAp) molecules sits on the crystallographic 3-fold axis. All four independent c-d(ApApAp) molecules have similar backbone conformations. The deoxyriboses are in the S-type pucker with pseudorotation angles ranging from 156.7 degrees to 168.6 degrees and the bases have anti glycosyl torsion angles (chi falling in two ranges, one at -104.3 degrees and the other ranging from -141.0 degrees to -143.8 degrees). In the R32 form, a hexahydrated cobalt(II) ion is found to coordinate through bridging water molecules to N1, N3, and N7 atoms of three adjacent adenines and oxygen atoms of phosphates. Comparison with other structures of cyclic oligonucleotides indicates that the sugar adopts N-type pucker in cyclic dinucleotides and S-type pucker in cyclic trinucleotides, regardless whether the sugar is a ribose or a deoxyribose. PMID- 9745897 TI - Molecular modeling of HIV-1 coreceptor CCR5 and exploring of conformational space of its extracellular domain in molecular dynamics simulation. AB - The chemokine receptor CCR5 functions as a major fusion coreceptor for macrophage tropic human immunodeficiency virus entry into cell. Here we report a three dimensional model of CCR5 built using molecular modeling approach. Because the virus binds to extracellular domain of the receptor, special attention was given to conformational flexibility, hydrogen bonding, and environmental polarity properties of this protein part. Such data were obtained in the result of molecular dynamics study of the extracellular domain. It was shown that during the simulation the extracellular segments form a compact globular domain with numerous long-range hydrogen bonds between them. First loop of the receptor stays quite rigid while N-terminal region and loops 2, 3 are rather flexible. A number of amino acid residues disposed in unfavourable environment and, therefore, potentially involved in binding of CCR5 to viral glycoproteins and chemokines, was delineated. Comparison of the results with available experimental data permits a proposal that such residues in loop-1 and N-terminal part of the receptor are important for HIV-1 entry, while those in loops 2 and 3 participate in ligand binding. Perspectives of rational alteration of virus-binding activity of CCR5 are discussed. PMID- 9745898 TI - Delineation of conformational preferences in human salivary statherin by 1H, 31P NMR and CD studies: sequential assignment and structure-function correlations. AB - Membrane-induced solution structure of human salivary statherin, a 43 amino acid residue acidic phosphoprotein, has been investigated by two-dimensional proton nuclear magnetic resonance (2D 1H NMR) spectroscopy. NMR assignments and structural analysis of this phosphoprotein was accomplished by analyzing the pattern of sequential and medium range NOEs, alphaCH chemical shift perturbations and deuterium exchange measurements of the amide proton resonances. The NMR data revealed three distinct structural motifs in the molecule: (1) an alpha-helical structure at the N-terminal domain comprising Asp1-Tyr16, (2) a polyproline type II (PPII) conformation predominantly occurring at the middle proline-rich domain spanning Gly19-Gln35, and (3) a 3(10)-helical structure at the C-terminal Pro36 Phe43 sequence. Presence of a few weak dalphaN(i,i+2) NOEs suggests that N terminus also possesses minor population of 3(10)-helical conformation. Of the three secondary structural elements, helical structure formed by the N-terminal residues, Asp1-Ile11 appears to be more rigid as observed by the relatively very slow exchange of amide hydrogens of Glu5-Ile11. 31P NMR experiments clearly indicated that N-terminal domain of statherin exists mainly in disordered state in water whereas, upon addition of structure stabilizing co-solvent, 2,2,2 trifluorethanol (TFE), it showed a strong propensity for helical conformation. Calcium ion interaction studies suggested that the disordered N-terminal region encompassing the two vicinal phosphoserines is essential for the binding of calcium ions in vivo. Results from the circular dichroism (CD) experiments were found to be consistent with and complimentary to the NMR data and provided an evidence that non-aqueous environment such as TFE, could induce the protein to fold into helical conformation. The findings that the statherin possesses blended solvent sensitive secondary structural elements and the requirement of non structured N-terminal region under aqueous environment in calcium ion interaction may be invaluable to understand various physiological functions of statherin in the oral fluid. PMID- 9745899 TI - The influence of pH alteration and pharmacological modulators of adenylate cyclase system on human serum albumin conformation. AB - The report describes the results of a study the effect of pH and binding of six physiologically active compounds (isoproterenol, yohimbine, theophylline, propranolol, clonidine and carbachol) on the molecular structure of human serum albumin (HSA) using dynamic light scattering. It was found that the albumin globule had the most compact configuration (Stokes diameter 59-62A) at physiological pH 7.4. The changes in pH both increased to 8.0 and decreased to 5.4, resulting in the growth of globule size to 72-81A. At acidic shift of pH an additional peak arose in the correlation spectra. This peak was caused by the light scattering on the structures with the Stokes diameters of 29-37A, which conformed to the sizes of the albumin subdomains. The additional peak was not displayed at basic shift of pH. The interaction with propranolol, clonidine and carbachol, which hinder adenylate cyclase (AdC) signaling system of a cell, initiated structural rearrangements similar to acidic transitions. Isoproterenol, yohimbine and theophylline, which activate AdC, caused the conformational changes of HSA similar to basic transitions. PMID- 9745900 TI - Simulated annealing for alpha-helical protein folding: searches in vicinity of the "molten globule" state. AB - A new model for simulation of protein folding of alpha-helical proteins with known secondary structure is proposed. We are dealing here with the analysis of alpha-helix packings rather than with a detailed atom structure of a whole protein. Starting from a random compact packing of the helices the search is focused on a vicinity of "molten globule" states of a protein. In contrast to the majority of the known approaches for estimation of a protein free energy we introduce a simplified potential of interactions with solvent and consider conformational energy of the loops in addition to mean-force potential. The model was applied to several globular alpha-helical proteins and demonstrated high prediction accuracy in comparison with other known models. PMID- 9745901 TI - Resonance Raman and infrared spectral studies on radical anions of model photosynthetic reaction center quinones (naphthoquinone derivatives). AB - Quinones play a vital role in the processes of electron transfer in bacterial photosynthetic reaction centers. It is of interest to investigate photochemical reactions involving quinones with a view to elucidate structure-function relationships in biological processes. Resonance Raman and FTIR spectra of electrochemically generated radical anions of 2-methyl-1,4-naphthoquinone, and 2 methyl-3-phytyl-1,4-naphthoquinone, also known as Vitamin K3 and Vitamin K1, respectively, (model compound for QA in Rhodopseudomonas viridis, a bacterial photosynthetic reaction center) have been reported. The same study has also been extended to 1,4-naphthoquinone for comparison. The vibrational assignments were carried out on the basis of comparison with our earlier time resolved resonance Raman studies on photochemically generated radical anions of 1,4-naphthoquinone and 2-methyl-1,4-naphthoquinone (Balakrishnan et al., J. Phys. Chem., 100, (1996), 16472-16478). These in vitro results have been compared with the reported vibrational spectral data under in vivo conditions. PMID- 9745902 TI - Dependencies of substitution steps number on Hamming distance are identical for one-parameter discrete models of both direct and parallel genetic diversity. AB - With the help of previously introduced enumeration procedure (M.Yu. Shchelkanov, A.N. Yudin, A.V Antonov, N.S. Starikov, A.A. Vedenov, E.V. Karamov, J. Biomol. Struct. Dyn. 15, 217-229 (1997)) and probability distribution function for the enumeration after some substitution steps (M.Yu. Shchelkanov, L.A. Soinov, V.V. Zalunin, D.A. Gumennyi, A.N. Yudin, A.A. Natan, V.B. Kireev, E.V. Karamov, J. Biomol. Struct. Dyn. 15, N 4, (1998)) we have demonstrated that dependencies of replication acts number on Hamming distance are identical for one-parameter discrete models of both direct and parallel genetic diversity. PMID- 9745903 TI - Stabilization of the purine.purine.pyrimidine DNA base triplets by divalent metal cations. PMID- 9745904 TI - Stabilization of the purine.purine.pyrimidine DNA base triplets by divalent metal cations. PMID- 9745905 TI - Neurotoxins and neurodegenerative disorders. AB - There is a hypothesis that excitotoxic pathomechanisms underlie neuronal tissue degeneration in neurological disorders. It is proposed that target neurons are overexcited, the result being energy disturbance and pathobiochemical changes that culminate in nerve cell death. Domoic acid (structural analogue of kainic acid) and beta-N-oxalylamino-L-alanine (BOAA) are powerful excitotoxins. Diminished energy supplies may result either in excessive release of glutamate or an inability of the neuron to restore ionic balance after stimulation. Impairment of intracellular energy metabolism induced by cyanide, amino-oxyacetic acid (AOAA), 3-acetylpiridine, thiamine deficiency, azide, 3-nitropropionic acid, methyl-phenyltetrahydro-pyridinum ion (MPP+), carbon monoxide, methanol intoxication and manganese (Mn) is discussed. Furthermore, the endogenous substance kynurenine is metabolised to kynurenic acid (excitotoxin antagonist) and quinolinic acid (excitotoxin agonist) Probenecid (inhibitor of kynurenic acid excretion from the extracellular fluid) has an antiexcitatory effect. PMID- 9745906 TI - The neurotoxicity of polychlorinated biphenyls. AB - Like dioxin, some polychlorinated biphenyl (PCB) congeners produce toxicity by binding to an aryl hydrocarbon (Ah) receptor. Other PCB congeners that have little or no activity at the Ah receptor have been shown to accumulate in the brain following in vivo exposure and decrease dopamine content. Subsequent research has found that non-dioxin-like PCBs also interfere with calcium homeostatic mechanisms and intracellular second messenger systems in vitro in neuronal cultures and brain subcellular fractions. The biological significance of these effects of PCBs in nervous system preparations is not known, although a number of calcium-dependent processes are important for nervous system function and development. Structure-activity relationship (SAR) studies based on measures of PCB-induced alterations in protein kinase C (PKC) translocation and Ca2+ buffering indicate that congeners with chlorine substitutions at the ortho position are active in vitro, while non-ortho congeners are relatively inactive. Subsequent research has found that chloride substitution patterns that favor non co-planarity are associated with activity in nervous system preparations. Recent in vivo studies in adults have shown that repeated exposure to a PCB mixture Aroclor 1254 increases translocation of PKC and decreases Ca2+-buffering in the brain. Increased levels of ortho-substituted non-coplanar PCB congeners were observed in the brains of Aroclor 1254-treated animals relative to vehicle controls. Current research is focusing on the possibility that PCB-induced alterations in calcium homeostasis and intracellular second messengers may be related to the developmental neurotoxicity of PCBs. PMID- 9745907 TI - Toxicokinetics and biochemistry of cadmium with special emphasis on the role of metallothionein. AB - Cadmium is a toxic metal with extremely long biological half-time of 15-20 years in humans. It has for decades been known that cadmium exposure can cause a variety of adverse health effects, among which kidney dysfunction, lung diseases, disturbed calcium metabolism and bone effects are most prominent. Following long term exposure the kidney is the critical organ. Cadmium and its compounds give rise to lung cancer after inhalation and have been classified as human carcinogens. Metallothionein (MT) is a low-molecular -weight protein, 6500Da with high cysteine content and high metal affinity, which plays a major role in the kinetics and metabolism of cadmium. The balance between CdMT and non-bound Cd in renal tissue has been shown to be of crucial importance for expression of toxicity. The most well studied metallothioneins are metallothioneins I and II with their isoforms which are expressed in almost all mammalian tissues. Metallothionein III is expressed in brain and is rich in zinc. Since the blood brain barrier keeps Cd outside the CNS, reported neurotoxic effects of Cd during development are likely to be secondary to an interference of Cd with Zn metabolism and not a direct effect of Cd on brain cells. It is therefore of importance to investigate whether neurotoxicity induced by cadmium is related to mechanisms involving MT III in brain. PMID- 9745908 TI - Myopathy: a possible effect of chronic low level lead exposure. AB - Morphological changes in the central nervous system and other organs have been reported in numerous studies investigating low level lead exposure. To date, however, there are no investigations on the effect of low level lead exposure on striated muscles, although varying neuromuscular changes in different species have been known for years. Rhesus monkeys were exposed pre- and postnatally to lead acetate in the diet (350 ppm or 600 ppm) over 9 years, followed by a lead free period of 32 months, while a control-group received regular diet. No signs of muscular dysfunction were evident. To elucidate neuromuscular pathomorphology frozen sections of the vastus medialis muscle were processed for routine and enzymohistological staining (Hematoxilin and Eosin, Sudan Black, Gomori, NADH, ATPase). Resin histology was processed for electron microscopy. Morphometric analysis was made with commercial software. Light microscopy revealed dose related signs of myopathy in the lead-exposed groups. The scatter of fibre diameters was increased, and split fibers and internal nuclei were more frequent. Fibres became separated from each other by copious endomysial connective tissue. Ultrastructural examination showed hydropic mitochondria and a massively dilated sarcotubular system in the 600 ppm group. Dose-related extracellular collagen deposition increased. A heavy fibrosis was seen in the 600 ppm group. These findings are interpreted as myopathical reaction due to chronic low level lead exposure, as there were no signs of neurogenical lesion. It remains unknown how the fibrosis developed. A primary fibrosis could be based upon a developmental delay of satellite cells (expressing metalloproteases for collagen-catabolism). Lead is known to inhibit regular development in many ways if exposure has started prenatally. As the skeletal muscle is a common target of toxicity, the myotoxic effects of chronic low level lead exposure comes into question. PMID- 9745909 TI - GFAP gene expression is altered in young rats following developmental low level lead exposure. AB - The astroglia cytoskeletal element, glial fibrillary acid protein GFAP, is a generally accepted sensitive indicator for neurotoxic effects in the mature brain. We used GFAP RNA as a marker for molecular biological changes in rat brain related to chronic low level lead exposure during the developmental period. Brains in the third week of postnatal life, following a continuous prenatal and postnatal exposure to lead of low dose were investigated. Sections were processed for in situ hybridization with a 35S labeled riboprobe, which is specific for mouse GFAP RNA. Radioactive signals which resulted from the RNA-RNA-hybridization process, were achieved by exposure of the sections to X-ray film. The gray values on the X-ray films were investigated by a computer assisted image analyzer system. Our data show that low level lead exposure affects the GFAP RNA-amount. Interestingly these effects were spatially diverse and different in the brain regions investigated. The gray values were significantly enhanced in the CA1 and CA2 regions of the hippocampus, in the striatum and in the cerebellum. The GFAP gene expression seems to reflect the reaction of the astroglia in accordance to the electrophysiological functions of the NMDA receptor in the diverse brain regions and their alterations following low level lead exposure. PMID- 9745910 TI - Comparative electrophysiological aspects of aluminum actions on central neurons and neuronal synapses of invertebrate and vertebrate animals. AB - Aluminum compounds (Al) increased the membrane potential (Em) and decreased the input resistance (Rin) of identified snail neurons. The neuronal excitability increased after Al withdrawal in the washing period. Cumulative Al applications caused dose-dependent modulation of Em and Rin in most of the studied neurons. Two phase actions were recorded on stimulus evoked excitatory postsynaptic potentials (EPSPs) or currents (EPSCs) at pH 6.5-6.9. The Al induced facilitation followed by attenuation were statistically significant, time- and dose-dependent events. They could be recorded at each Em. Low affinity and slow binding kinetics could characterize the Al actions on the neurons. Al showed pH- dependent suppression of EPSPs or EPSCs in some neurons. Our findings are partially comparable with Al induced electrophysiological and pharmacological modifications reported on vertebrate neurons. PMID- 9745911 TI - The role of iron in neurotoxicity: a study of novel antimalarial drugs. AB - The antimalarial drug artemisinin and its derivatives display neurotoxicity in animal studies in vivo and in neuronal cells in vitro. Their toxicity may be due to an interaction of iron with the endoperoxide bridge of the derivative to produce toxic free radicals and/or other toxic metabolites. In this study, 0.3 microM artemether (AEM) in the presence of 2 microM haemin significantly inhibited the outgrowth of neurites from differentiating NB2a neuroblastoma cells by up to 76%. The antioxidants ascorbic acid and glutathione completely protected against this toxicity at a concentration of 100 microM. AEM was found to be partially converted to two isomeric products, which were identified as the tetrahydrofuran acetate isomer of AEM and 3alpha-hydroxydesoxyartemether. PMID- 9745912 TI - Vanadium as a factor that disturbs phosphorus metabolism in nervous tissue. AB - Vanadium and its derivatives are the well-known environmental pollutants. We obtained that ammonium vanadate suppressed the alkaline (AlP) and acid (AcP) phosphatases activity in the variety of tissues. Vanadate inhibited the enzymes that take part in phosphoryl transfer reactions. Successive concentration dependent decrease in AlP and AcP activities by vanadate (0.1 mM-1 mM) has been obtained in rat nervous tissue. Moreover, the observed reduction in the sensitivity of the enzymes to vanadate ions becomes more pronounced at the transition from higher and relatively young regions of nervous system to its more ancient parts. Phosphoprotein phosphatase (PP) activity has also been inhibited in various structures of brain tissue; in spinal cord, however, vanadate in comparatively low concentrations (10 microM) caused a steep rise in the enzyme activity. Only at high concentration (1 mM) vanadate exert the effect of moderate inhibition. The possibility of the existence of phylogenetically different molecular forms of nervous tissue PPs differed by their sensitivity to vanadate was supported in our comparative examination. PMID- 9745913 TI - Pharmacological actions of essential (zinc, copper) and toxic metal ions (lead, mercury) on neurons and neuronal synapses of the snail, Helix pomatia L. AB - Actions of zinc, copper, lead and mercury ions on identified neurons and neuronal synapses of the snail, Helix pomatia L. were examined by use of electrophysiological methods. The studied metal ions depolarized the membrane and decreased the resistance but the neuronal excitability changed differentially. Copper and zinc transiently increased the amplitude and/or frequency of the spontaneous EPSPs or EPSCs in time- and dose-dependent ways. Lead attenuated the evoked EPSPs or EPSCs but copper and in some neurons zinc and mercury induced dose-dependent biphasic effects. Metal ions influenced the potential dependence, reversal potential, rate of rise and rate of fall of evoked responses differentially. The actions of zinc and lead on evoked EPSPs were reversible but those of copper and mercury were not and DTT treatment was ineffective. The affinity of the metal ions to the binding site(s) was low and the binding kinetics slow. The sensitivity of the binding site decreased in the order of mercury>copper>lead>zinc according to the IC50 values. The binding stoichiometry (nH value) changed from 1.4 to 2.6 for the various metal ions. PMID- 9745914 TI - Polychlorocycloalkane insecticide action on GABA-and glycine-dependent chloride flux. AB - The inhibitory neurotransmitters gamma-aminobutyric acid (GABA) and glycine directly cause an increase in conductance to Cl- by binding to ligand-operated ion channel receptors at the postsynaptic membranes, so that opening of Cl- channels usually leads to a net hyperpolarization. The GABA(A) receptor has separate but allosterically interacting binding sites for GABA, benzodiazepines, barbiturates, anesthetic steroids and the convulsant picrotoxinin. The GABA(C) receptor also forms a Cl- channel, however its pharmacology differs from that of the GABA(A) receptor. Neurotoxic organochlorine pesticides belonging to the group of polychlorocycloalkanes (cyclodienes and gamma-hexachlorocyclohexane or lindane) induce in mammals an hyperexcitability syndrome that can progress until the production of tonic-clonic convulsions. They act as non-competitive GABA antagonists interacting with the picrotoxinin site both in membranes and in intact cultured neurons, thereby inhibiting the GABA-induced Cl- flux following activation of either GABA(A) or GABA(C) receptors. We also report the effects of polychlorocycloalkanes on glycine-induced 36Cl- flux in primary neuronal cultures. The delta isomer of hexachlorocyclohexane is a depressant compound, that increases the GABA-induced Cl- flux and allosterically increases benzodiazepine binding at the GABA(A) receptor. We discuss the mechanism of action of these compounds in relation to the disruption of ligand-operated Cl- channel receptors and the relevance of their convulsant/depressant actions. PMID- 9745915 TI - Ion channels as targets for insecticides. AB - Most insecticides are neurotoxicants causing various forms of hyperexcitation and paralysis in animals. A variety of neuroreceptors and ion channels have been identified as the major target sites of these neurotoxic insecticides. This paper gives the highlights of some of the recent development in this area. Pyrethroids keep the sodium channel open for unusually long times causing a prolonged flow of sodium current. The prolonged sodium current elevates and prolongs the depolarizing after-potential which reaches the threshold membrane potential to initiate repetitive after-discharges. We have developed the method with which the percentage of sodium channel population that needs to be modified to cause repetitive after-discharges can be measured accurately. In rat cerebellar Purkinje neurons, only 0.6% of sodium channels needs to be modified for hyperexcitation resulting in a large toxicity amplification. This concept is applicable to other neuroactive drugs that act through the threshold phenomenon. The mechanisms of selective toxicity of pyrethroids in mammals and insects have been quantitatively determined to be due mainly to the different sensitivity of the sodium channels to pyrethroids and the negative temperature dependence of pyrethroid action on the sodium channels. The degradation of pyrethroids play only a minor role. The negative temperature dependence of pyrethroid action is due to the increased sodium current flow at low temperature. The major site of action of dieldrin and hexachlorocyclohexane is the GABA(A) receptor chloride channel complex. Dieldrin exerts a dual action, initial stimulation and subsequent suppression, and the latter is responsible for hyperexcitation of animals. Dieldrin stimulation requires the gamma2s subunit in the GABA receptor, whereas dieldrin suppression occurs in the presence or absence of the gamma2s subunit. PMID- 9745916 TI - Synergism between NMDA and domoic acid in a murine model of behavioural neurotoxicity. AB - We have examined the behavioural neurotoxicity of domoic acid (DOM) and kainic acid (KA) in mice following administration of ligands active at the N-methyl-d aspartate (NMDA) receptor. Groups of female CD-1 mice (n=4) were injected i.p. with saline or one of three doses of either DOM or KA. Doses of DOM and KA were selected from the steep portion of the respective dose response curves and were equitoxic when compared between the two ligands. Toxicity was recorded as both total cumulative toxicity over 60 min according to a previously validated 7 point rating scale, and as the latency to the onset of tremors and/or convulsions. Five minutes prior to administration of either agonist mice were injected with either saline, NMDA (40 mg/kg) or a combination of NMDA and 15 mg/kg CPP (3-[2 carboxypiperazine-4-yl]propyl-1-phosphonic acid). Neither NMDA nor CPP at these doses produced significant changes from baseline responding when injected prior to saline. Injection of NMDA prior to DOM, however, resulted in significantly increased cumulative toxicity and significantly reduced latencies to seizures at the two highest doses of DOM (3.75 and 5.0 mg/kg). NMDA-induced potentiation of DOM toxicity was completely antagonized by co-administration of CPP. In contrast, injection of NMDA prior to KA did not result in significant changes in KA toxicity at any of the doses tested using either index of behavioural toxicity. These results confirm previous reports of synergism between DOM and ligands acting at the NMDA receptor in isolated neurons, and provide further evidence of pharmacological dissociation of the actions of DOM and KA in vivo. PMID- 9745917 TI - In vitro effect of the cysteine metabolites homocysteic acid, homocysteine and cysteic acid upon human neuronal cell lines. AB - Cysteine (CYS) is a non-essential amino acid which elicits excitotoxic properties via the N-methyl-D-aspartate (NMDA) subtype of the glutamate receptor. CYS levels are known to be elevated in association with neurological disease such as Alzheimers Disease (AD) and Parkinsons Disease (PD). We have previously reported studies investigating the toxicity of CYS and its major metabolite cysteinesulfinic acid (CSA) to human neuronal cell lines in vitro and in continuation of this we now report the toxicity of other compounds (Homocysteic Acid, HCA; Homocysteine, HCYS; and Cysteic Acid, CA) in the CYS metabolic pathway. Three cell lines, all of human origin and derived from separate discrete areas of the brain were used in the neurotoxicity assays. Lactate dehydrogenase (LDH) release was assayed as a measure of cell death. The cell lines investigated showed varying degrees of toxic responses which were the reverse of those seen when they were exposed to CYS or CSA. The SK.N.SH (Neuroblastoma) cell line, which exhibits a high toxic response to CYS and CSA, gave a low toxic response to HCA and CA while the TE 671 (Medulloblastoma) cell line, which exhibits a low toxic response to CYS and CSA, showed a high toxic response to HCYS, HCA and CA. However, the U-87 MG (Glioblastoma) cell line, which has a median toxic response to CYS and CSA, also has median response to HCYS, HCA and CA. These results show that toxic responses are cell-type specific for CYS and its metabolites and this may be reflected in the patterns of neurodegeneration observed in such diseases as AD and PD. HCYS is selectively toxic to medulloblastoma cells; this may explain why high HCYS levels result in neural tube defects in prenatal humans, where the same cell-type is involved. PMID- 9745918 TI - Insect (Locusta migratoria migratorioides) test monitoring the toxicity of cyanobacteria. AB - An insect test was developed to investigate the toxicity of cyanobacteria. The African locust, Locusta migratoria migratorioides R.F. was used as a test animal instead of mouse. The cyanobacteria tested were Aphanizomenon flos-aque, Anabaena aphanizomenoides, Cylindrospermopsis raciborskii, Microcystis aeruginosa. The toxicity of authentic microcystin-LR was also tested. Cyanobacteria producing toxins killed the animals when the homogenized cell suspension was injected into the animals. The locust test proved to be more sensitive than the mouse test. The LD50 values of the different cyanobacteria for locusts and for mice, respectively were the following: 90 microg/animal (60 mg/kg) and 8000 microg/animal (320 mg/kg), for Aphanizomenon flos-aquae; 255 microg/animal (170.2 mg/kg) and 3750 microg/animal (150 mg/kg), for Anabaena aphanizomenoides; 195 microg/animal (131.4 mg/kg) and 5750 microg/animal (230 mg/kg), for Cylindrospermopsis raciborskii; 22.5 microg/animal (15 mg/kg) and 6000 microg/ animal (240 mg/kg), for Microcystis aeruginosa. In locusts the LD50 value for authentic microcystin LR was 0.2 microg/animal (130 mg/kg). Since the weight of the mice is 15 to 20 times larger than that of the locusts, hence less toxic cells are needed to kill the locusts. The locust test is cheaper than the mouse test, large number of animals can be used in the experiments and the LD50 values can be estimated more precisely. The toxicity of C. raciborskii was significantly lower when the lyophilized cells were extracted in methanol (LD50 = 767 mg/kg), instead of NaCl solution (LD50 = 131.4 mg/kg). PMID- 9745919 TI - Experimental model studies of pesticide exposure. AB - The neurotoxic effects of Dimethoate (Dim), Dichlorvos (DDVP) and Methyl Parathion (MP) respectively were investigated on the central nervous system (CNS) and peripheral nervous system (PNS) of rats after different treatment schedules at the macro and single unit cell level. At the macro investigations 1/25, 1/50 and 1/100 of the respective LD50 values of each pesticide were administered to different groups by gavage daily in the following programs: Pregnancy variation (P) to females from 5th to 15th days of pregnancy; Pregnancy and lactation variation (P+L): to females as above and during lactation for 4 weeks; Pregnancy+lactation+post weaning variation (P+L+P) as above plus to the young male rats (F1 generation) up to 8 weeks. Neurotoxicological investigations were conducted on the F1 rats at the age of 12 weeks. Spontaneous electrocorticograms (ECoG) were recorded on the anesthetized rats from the somatosensory, visual and auditory cortex. Cortical evoked potentials (EP) were recorded from the same areas subsequently. Conduction velocity and refractory periods of the tail nerve was investigated. Treatment by Dim, DDVP and MP during P and P+L of the mothers did not influence the bioelectric activity of the offsprings significantly. The same treatment by the P+L+P programme, resulted in significant changes. Frequency of the spontaneous ECoG waves grew significantly in all dose groups of P+L+P group. Latency time become shorter after somatosensory. light or acoustic stimuli respectively on one hand and the beginning of the of answer of these by the evoked potential (EP) waves on the other hand. Conduction velocity of the tail nerve diminished, refractory periods grew dose dependently and significantly at the P+L+P programs with all the three pesticides. Cortical single unit activity was studied after the i.p administration of 1/5 LD50 of the three organophosphates (OP). The decrease of the firing frequencies was observed. The amplitude of the hippocampal population spikes increased. The changes observed in these studies point toward a higher excitation state of the CNS and a disturbed conduction of the nervous impulses of the peripheral nerves. These results could be taken into consideration when deciding on human contaminations by OP-s. PMID- 9745920 TI - Developmental neurotoxicological effects of lead and dimethoate in animal experiments. AB - Neurophysiological changes caused by parallel treatment with inorganic lead and dimethoate (a combination of possible health risk at population level) were investigated in different phases of the ontogenesis. Wistar rats were treated by gavage with lead (80.0 or 320.0 mg/kg); with dimethoate (7.0 or 28.0 mg/kg), or with their combination on days 5-15 of pregnancy, days 5-15 of pregnancy + days 2 28 of lactation (females of P generation), or days 5-15 of pregnancy + days 2-28 of lactation (females of P generation) + 8 weeks after weaning (males of F1 generation). Electrophysiological parameters (electrocorticogram, cortical evoked potentials) of the F1 male rats in the above groups were investigated at the age of 12 weeks. Both spontaneous and evoked electrophysiological phenomena showed dose-, treatment- and combination-dependent changes (e.g. significantly decreased mean amplitude and increased frequency of the electrocorticogram, lengthened latency and duration of the somatosensory, visual and auditory evoked potentials) which seemed to be more pronounced in the groups treated with the combination of lead and dimethoate than in the groups given lead or dimethoate alone. The results indicate that a simultaneous, pre- and postnatal exposure to the neurotoxicants, lead and dimethoate, considerably altered the functioning of the central nervous system. PMID- 9745921 TI - Comparative studies of O,O-dialkyl-O-chloromethylchloroformimino phosphates: interaction with neuropathy target esterase and acetylcholinesterase. AB - Acetylcholinesterase (AChE) and neuropathy target esterase (neurotoxic esterase, NTE) are two major target enzymes for organophosphorus (OP) esters. The relative potency of an OP ester to react with AChE or with NTE in vitro correlates with its relative potency in vivo to cause acute toxicity (death) or organopohosphate induced delayed neurotoxicity (OPIDN). On this basis extrapolation from in vitro to in vivo data now seems justifiable to predict risk of OPIDN. The kinetics of NTE and AChE inhibition by experimental pesticides of the general formula (RO)2P(O)ON=CClCH2Cl, where R = methyl, ethyl, isopropyl, propyl, isobutyl, butyl, pentyl, has been studied. Compounds with short R (methyl, ethyl) were shown to be far more potent inhibitors of AChE than NTE. Both anti-NTE activity, selectivity for NTE and, correspondingly, the propensity of compounds to cause OPIDN rise with increasing their hydrophobicity. A high value of ki(NTE)/ki(AChE) for R = pentyl suggests that this compound would have the potential to cause OPIDN at doses lower than the LD50. A quantitative structure-activity relationships (QSAR) analysis indicated that NTE and AChE have different structural and electronic requirements for their respective OP inhibitors. PMID- 9745922 TI - The effect of benomyl on neurite outgrowth in mouse NB2A and human SH-SY5Y neuroblastoma cells in vitro. AB - The commercial fungicide methyl 1-[(butylamino) carbonyl]-1H-benzimidazol-2 ylcarbamate (benomyl) is teratogenic in rats. Its mode of action is believed to be related to its ability to inhibit the polymerization of brain tubulin. In this study its effects were studied in cultured neuronal cells during differentiation and neurite outgrowth. Mouse NB2a and human SH-SY5Y neuroblastoma cells were induced to differentiate by addition of dibutyryl cyclic AMP and at the same time were exposed to various concentrations of benomyl. Benomyl significantly inhibited neurite outgrowth in both cell lines at concentrations of 10(-8) M and above with IC50 values of 5.9 x 10(-7) M and 1.0 x 10(-6) M in the NB2a and SH SY5Y cells respectively. The results show that benomyl inhibits neuronal cell differentiation at concentrations likely to be achieved during the development of fetal abnormalities in rats in vivo. PMID- 9745923 TI - Genetic polymorphisms in Parkinson's disease. AB - The search for genetic polymorphisms relevant to Parkinson's disease etiology and pathogenesis has been motivated by recent thinking emphasizing the potential significance of gene-environment interactions. Especially influential to this research have been the MPTP model of PD induction, hypotheses concerning oxidative stressor reactions, and epidemiological observations of an inverse relation between cigarette smoking and PD risk. This brief review summarizes trends in genetic polymorphism research, with examples provided by investigations of cytochrome P450 enzymes, monoamine oxidase, superoxide dismutase, and mitochondrial genes. PMID- 9745924 TI - Genetically determined susceptibility to organophosphorus insecticides and nerve agents: developing a mouse model for the human PON1 polymorphism. AB - Several organophosphorus insecticides and nerve agents are detoxified through the cytochrome P450/paraoxonase (PON1) pathway. PON1 is an HDL-associated enzyme encoded as a 355 amino acid protein in humans. The PON1 Arg192 isoform hydrolyzes paraoxon rapidly while the Gln192 isoform hydrolyzes this compound slowly. Both isoforms hydrolyze phenylacetate and chlorpyrifos oxon at approximately the same rate. We recently found that the effect of this polymorphism is dramatically reversed for sarin hydrolysis. The PON1 Arg192 isoform has virtually no sarinase activity while the Gln192 isoform has substantial activity. The Gln192 isoform also hydrolyzes diazoxon and soman faster than the Arg192 isoform. In addition to the large differences in rates of hydrolysis observed for some OP substrates by the two PON1 isoforms, there is also a large variability in serum PON1 concentrations that is stable over time between individuals. Thus, two factors govern the PON1 status of a given individual, the PON1 genotype as well as the amount of protein expressed from each allele. A two-dimensional enzyme analysis provides an excellent assessment of an individual's PON1 status, ie. the position 192 genotype as well as phenotype, or level of serum PON1 (Nature Genet 14:334 336). Do these interindividual differences in rates of substrate hydrolysis by PON1 reflect an individual's sensitivity or resistance to OP compounds processed through the P450/PON1 pathway? Injection of purified PON1 into mice clearly demonstrates the protective effect of having high serum levels of PON1 against toxicity by chlorpyrifos oxon or chlorpyrifos. Preliminary experiments with PON1 knockout mice, on the other hand, clearly demonstrate that low PON1 levels result in dramatically increased sensitivity to chlorpyrifos oxon. Attempts to express human PON1 in mice from constructs containing either of the human PON1 cDNA sequences were unsuccessful, despite the generation of the respective transgenic mice. PMID- 9745925 TI - Metallothionein (MT) isoforms in the central nervous system (CNS): regional and cell-specific distribution and potential functions as an antioxidant. AB - Similar to the "housekeeping" functions ascribed to MTs in other tissues, central nervous system (CNS) metallothioneins (MTs) are implicated in metal metabolism, cellular repair processes, growth and differentiation, where they are likely to serve as the source of zinc for newly synthesized apoenzymes, as well as regulator molecules in gene expression. Additional likely functions of MTs include control of intracellular redox potential, and metal detoxification. This manuscript will focus on (1) the distribution of MT isoforms within the CNS, with particular emphasis on the cell-specific localization of MTs and its significance, (2) emerging accounts on the function of MT as an antioxidant, and (3) the relationship between MT and ethanol (EtOH)-induced neurotoxicity in a model system of neonatal rat primary astrocyte cultures. PMID- 9745926 TI - Regulation of the synthesis of brain metallothioneins. AB - Metallothioneins (MTs) area family of low molecular weight proteins characterized by a high cysteine (about 30%) and heavy metal (Zn2+, Cu+) content. In rodents, there are four known MT isoforms, named MT-I to MT-IV. MT-I and MT-II are two widely expressed isoforms, while MT-III and MT-IV (isoforms recently discovered) have a more restricted expression, normally in the brain and in the keratinizing epithelia, respectively. Since all MT isoforms share a substantial homogeneity regarding their heavy metal binding properties, it seems feasible that they could also share physiological functions. However, the different pattern of expression suggest that the different MT isoforms could in addition have specific roles. Indeed, MT-III (or GIF) was initially discovered as a protein with apparent neuromodulatory effects, which were not shared by the normal counterparts MT-I and MT-II. To gain insight on the putative importance of these three MT isoforms in the brain, it is important to characterize their regulation in normal and pathophysiological states. In this review we summarize the major factors known to affect brain MT regulation. PMID- 9745927 TI - Glutamate-stimulated ROS production in neuronal cultures: interactions with lead and the cholinergic system. AB - Oxidative stress may be an important factor in several pathological brain conditions. A contributing factor in many such conditions is excessive glutamate release, and subsequent glutamatergic neuronal stimulation, that causes increased production of reactive oxygen species (ROS), oxidative stress, excitotoxicity and neuronal damage. Glutamate release is also associated with illnesses such as Alzheimer's disease, stroke, and brain injury. Glutamate may interact with an environmental toxin, lead, and this interaction may result in neuronal damage. Glutamate-induced ROS production is greatly amplified by lead in cultured neuronal cells. Alterations in protein kinase C (PKC) activity seem to be important both for glutamate-induced ROS production, and for the amplification of glutamate-induced ROS production by lead. It is possible that the neurotoxic effects of lead are amplified through glutamate-induced neuronal excitation. Cholinergic stimulation can also trigger ROS production in neuronal cells. PKC seems to play a key-role also in cholinergic-induced ROS production superoxide anion being the primary reactive oxygen species. There seems to be a close relationship between the responses of cholinergic muscarinic and glutamatergic receptors because glutamate receptor antagonists inhibit cholinergic-induced activation of human neuroblastoma cells. Glutamatergic neuronal stimulation may be a common final pathway in several brain conditions in which oxidative stress and ensuing excitotoxicity plays a role. PMID- 9745928 TI - Neurotoxicity of ammonia and glutamate: molecular mechanisms and prevention. AB - Ammonia is a main factor in the pathogenesis of hepatic encephalopathy. We found that acute ammonia toxicity is mediated by activation of NMDA receptors. Chronic moderate hyperammonemia prevents acute ammonia toxicity in rats. Chronic exposure of cultured neurons to 1 mM ammonia leads to impaired response of the NMDA receptor to activation by its agonists (due to decreased protein kinase C mediated phosphorylation) and prevents glutamate (Glu) neurotoxicity. Compounds that prevent ammonia toxicity in mice (e.g. carnitine) also prevent Glu toxicity in cultured neurons. These compounds did not prevent activation of NMDA receptor or the rise of Ca2+. They interfered with subsequent steps in the toxic process. The protective effect of carnitine is mediated by activation of metabotropic Glu receptors. Agonists of mGluRs, especially of mGluR5, prevent Glu toxicity. Agonists of muscarinic receptors also prevent Glu toxicity and there seems to be an interplay between muscarinic and metabotropic Glu receptors in the protective effect. We have tried to identify intracellular events involved in the process of neuronal death. It is known that the rise of Ca2+ is an essential step. Glu leads to depletion of ATP; some compounds (e.g. carnitine) prevent Glu-induced neuronal death without preventing ATP depletion: additional events are required for neuronal death. Glu induces activation of Na+/K+-ATPase, which could be involved in the toxic process. Inhibitors of protein kinase C, calcineurin or nitric oxide synthase prevent Glu toxicity. Our results indicate that Glu toxicity can be prevented at different steps or by activating receptors coupled to the transduction pathways interfering with the toxic process. Agents acting on these steps could prevent excitotoxicity in vivo in animals. PMID- 9745929 TI - Mechanisms of prionSc- and HIV-1 gp120 induced neuronal cell death. AB - In vitro experiments revealed that the scrapie prion protein, PrP(Sc), as well as the PrP fragment PrP106-126, and the HIV-1 coat protein gp120 induce apoptosis of rat cortical neurons. The toxic effect displayed by PrP and gp120 could be blocked by NMDA receptor antagonists. Treatment of neuronal cells with PrP106-126 resulted in a drop of intracellular glutathione level and changes in the level of Bcl-2. Evidence is presented that gp120 causes an activation of phospholipase A2, resulting in the increased release of arachidonic acid, which may in turn sensitize the NMDA receptor. PMID- 9745930 TI - Preliminary evidence of neurotoxicity associated with eating fish from the Upper St. Lawrence River Lakes. AB - Pollution of hydrographic basins has affected the flora and fauna that thrive in these aquatic ecosystems, and fish, which constitute an important food resource, often contain a plethora of potentially toxic chemicals. In a major research project on early neurotoxic effects of environmental exposure to manganese among residents in Southwest Quebec, fish consumption from 2 lakes of the Upper St. Lawrence River System, was surveyed as a potential confounding factor. Participants were selected using a random, stratified sampling strategy from lists of the Quebec Health Plan. Following exclusions, 273 men and women between 20-69 years were retained for the present analysis. A total of 103 (37.7%) reported eating fish from the Upper St. Lawrence. Although fisheaters and non fisheaters were similar for most socio-demographic variables, significantly more fisheaters (65.2%) reported consuming alcoholic beverages as compared to non fisheaters (42.4%) (Chi Sq. <0.01). To eliminate this possible bias, fisheaters were matched to non-fisheaters for the variables sex, alcohol consumption (never or occasionally vs. regularly), age (+/-5y) and education (+/-2y). A total of 63 matched pairs were thus created. Paired analyses (t-test or Signed Rank) showed that fisheaters had higher levels of blood organic mercury and lead. Analysis of nervous system functions revealed that both groups performed similarly on tests of sensory function, visual memory and recognition, fine motor performance and some motor tests, but fisheaters performed significantly more poorly (p<0.05) on tests requiring cognitive flexibility, word naming, auditory recall, and more complex motor tasks. The profile of deficits is consistent with diminished capacity for information processing. These observations were made within a study that was not specifically designed to examine the effects of fish eating from these two lakes, and the characterization of fish dietary habits has many limitations. Nevertheless, the findings are sufficiently compelling to warrant further studies, since fish from the Upper St. Lawrence Lakes are known to contain multiple neurotoxic substances. PMID- 9745931 TI - Predictive validity of the Q16 questionnaire: a comparison between reported symptoms and neurobehavioral tests. AB - The correspondence between the answers to the Q16 questions regarding memory and attention-concentration and relevant neurobehavioral performance test scores has been evaluated. The sensitivity, specificity and diagnostic validity of Q16 have been assessed, taking the relevant neurobehavioral test score as a reference diagnostic criterion, the lower quartile of performance being considered as a poor response. The group under study consisted of 74 volunteers (24 females), aged 40 years on average (SD:7.5) and recruited among styrene-exposed workers and healthy controls. The test battery included the logical memory (short- and long term) and the verbal learning (short- and long-term) tests of the Wechsler Adult Intelligence Scale (WAIS). The answers to the Q16 questions were poorly related to the performance: self-perceived forgetfulness showed a limited agreement with the long-term logical memory test (r=-0.23, p<0.05). The number of false negatives (no symptom but low test scores) was generally high, giving rise to a very low sensitivity of the questionnaire, despite a relatively high specificity. Accordingly, the positive diagnostic validity was low (<30%), whereas the negative diagnostic validity was high (>80%). Different methods used to investigate subtle neurological changes give rise to inconsistencies between self perceived disturbances and objective measurements of relevant functions. Owing to its low sensitivity and positive diagnostic value, the Q16 cannot be recommended as a screening tool among workers occupationally exposed to neurotoxic chemicals. PMID- 9745932 TI - A case-control study of occupational and environmental risk factors for Parkinson's disease in the Emilia-Romagna region of Italy. AB - A questionnaire-based case-control study was carried out on 86 patients with neurologist-confirmed idiopathic Parkinson's disease (PD) and 86 controls similar in sex and age. The control group was recruited in outpatient specialist centers of the same University Hospital (glaucoma, psoriasis vulgaris, essential arterial hypertension and renal diseases). Exposure was defined as occupational or residential contact with a given factor for at least 10 consecutive years prior to the onset of PD. Smoking habits were defined by exclusion of those subjects who never smoked. The following risk factors were identified: cranial trauma (OR: 2.88; 95% CI: 0.98-8.49), well water use (OR: 2.78; 95% CI: 1.46-5.28) and occupational exposure to industrial chemicals (OR: 2.13; 95% CI: 1.16-3.91). Among industrial chemicals, only organic solvents were identified as significant risk factors for PD (O.R. : 2.78, 95% C.I. : 1.23-6.26). Whereas no exposure to neurotoxic metals occurred among controls, making the assessment of the O.R. impossible, exposure pesticides and herbicides was similar in the two groups (O.R. : 1.15; 95% C. : 0.56-2-36). Smoking habits was negatively associated with PD (OR: 0.41; 95% CI: 0.22-0.75), confirming the "protective" role of tobacco smoking suggested by many studies. As a whole, these results support the role of environmental factors in the etiology of PD. PMID- 9745933 TI - Association of biochemical and subjective indicators of drinking habits with performance on different neurobehavioral tasks. AB - The present paper outlines the association of biochemical and subjective indicators of alcohol consumption. Due to its relevance as a potential confounding variable in occupational neurotoxicology, both sources of information about drinking habits were related to neurobehavioral test performance. A sample of 308 rotogravure printers and control subjects from a cross-sectional longitudinal study in various German printing plants was studied. Duration of employment was 4 months to 44 years (mean = 14.9, sd = 9.67). Mean age was 38.4 years (range 21 - 60). From venous blood samples three parameters considered to be sensitive for increased consumption of alcohol were used. They were carbohydrate-deficient transferrin (CDT), gamma glutamyl transferase (GGT), and mean cell volume (MCV). During the medical interview subjects with any chronic liver disease were identified and excluded from data analysis. Additionally, information about weekly consumption of alcohol was assessed and transformed to grams per day (g/d) values. Neurobehavioral testing included simple reaction time (SPES version), switching attention, symbol digit substitution, and digit span (EURONEST version). Additionally, a questionnaire of neurotoxic complaints was administrated. Other covariates, i.e. verbal ability, history of solvent exposure, and age were controlled. GGT and CDT were elevated in 10.5% and 6.6% of the population. 3.5% of the subjects reported daily consumption higher than 60 gram. There were positive correlations of CDT and GGT with the subjective indicator of drinking habits. The magnitude of these relationships were low, but the associations were significant. MCV was not correlated with subjective reports of drinking habits, but it showed convergent correlations with CDT and GGT. Comparison of these two parameters with performance on neurobehavioral tasks yielded only one negative association, i.e. between the memory-loaded tasks factor and GGT. CDT and subjective estimation of alcohol consumption were not related to any cognitive function tested in this study. Especially, the digits backward task was negatively correlated with increased GGT. PMID- 9745934 TI - Toxicity of the styrene metabolite, phenylglyoxylic acid, in rats after three months' oral dosing. AB - Male Wistar rats were dosed with 0, 1250, 3750 or 5000 mg/l of phenylglyoxylic acid (PGA) (CAS no. 611-73-4) in the drinking water ad libitum for 3 months. During the entire treatment period, there were no gross signs of toxicity related to PGA. No changes in neurobehavior were found after using a functional observational battery or radial arm maze. An increased relative kidney weight was seen in the highest dose-group (Controls: 0.504 +/- 0.031 g/100 g b.wt.; 5000 mg PGA/l: 0.579 +/- 0.033 g/100 g b.wt.; p<0.01). No other organ weights were affected. Histopathology revealed no change in kidney structure. No changes in clinical biochemistry. In the highest dose-group three animals out of ten showed reduction in peripheral nerve myelin sheath thickness. No such changes were seen in the control group. The study revealed no changes in auditory brain stem response but minor changes in electroretinography. The noradrenaline (NA) concentration decreased in pons and thalamus whereas it increased in medulla oblongata and whole brain. The dopamine (DA) concentration increased in cerebellum, hippocampus, pons, and whole brain. The most marked DA increase was seen in hippocampus (Controls: 0.56 +/- 0.10 nmol/g tissue; 5000 mg/l: 1.04 +/- 0.11 nmol/g tissue; p<0.001). The 5-hydroxytryptamine (5-HT) concentration decreased in cerebellum, cerebral cortex, hippocampus, and medulla oblongata, whereas it increased in thalamus. The yield of synaptosomal protein, synaptosomal NA, DA, and 5-HT concentrations, and DA uptake rate were not affected. When dosed males were mated with naive females, there were no differences between groups in the pregnancy rate, number of corpora luteae, implantations, live or dead fetuses, resorptions, preimplantation loss, or postimplantation loss. It is concluded that a part of the effects on kidney, peripheral nerves, and vision, which have previously been reported after exposure to styrene, might be induced by the styrene metabolite, PGA. If PGA has ototoxic effects in rats, the dosing in the present study is not sufficient to induce the necessary ototoxic concentration in blood. Alternatively, the ototoxicity of styrene, like toluene, may be caused the parent compound itself and not by a metabolite like PGA. PMID- 9745935 TI - Decreases in brain glial fibrillary acidic protein (GFAP) are associated with increased serum corticosterone following inhalation exposure to toluene. AB - Toluene and other neurotoxicants can cause both increases and decreases in the concentration of GFAP in the brain. While increased GFAP concentration is widely regarded as evidence for reactive gliosis, toxicant-induced decreases in GFAP have received less attention. In order to identify conditions under which inhalation exposure to toluene results in decreased GFAP concentration, rats were subjected to repeated inhalation of toluene for up to 7 days. Adult male F344 rats received inhalation exposure to air or to 1000 ppm toluene, 6 hr/day, for 3 or 7 days. This toluene exposure replicated the previously-observed decreases in GFAP in the thalamus. Serum Corticosterone was significantly elevated in the same rats that exhibited decreases in brain GFAP concentration. These results show that decreases in brain GFAP might be a consequence of disruption of the hypothalamic-pituitary-adrenal axis and/or hormonal homeostasis. Changes in GFAP and in Cort were not accompanied by a change in body weight. More research is needed to firmly establish cause and effect between increased serum glucocorticoid levels and GFAP decreases following toluene inhalation and to determine whether these decreases indicate toxicity or adaptive changes. PMID- 9745936 TI - Digital radiotherapy simulator. AB - We describe a prototype digital radiotherapy simulator which consists of a conventional simulator gantry, digital spot imager, and image correction and reconstruction software. The ability of the digital spot imager to acquire a diagnostic quality image directly in digital format during simulation offers unique possibilities in clinical practice. Applications include prescription of multileaf collimator, on-line patient setup verification, remote consultation and treatment planning. In addition, we discuss the possibility of using the digital simulator as a volume-CT scanner capable of obtaining three-dimensional anatomical information in a single scan. PMID- 9745937 TI - Intraluminal signal intensity of iliac artery stents investigated by contrast enhanced three-dimensional MR angiography. AB - The purpose of this study is to investigate the intraluminal signal of iliac artery stents by contrast-enhanced three-dimensional magnetic resonance (MR) angiography with a short echo time. Ten patients with iliac arterial diseases treated by intravascular metallic stent placement were examined with a 1.5 T MR imager. Gadolinium-enhanced MR angiography depicted the intraluminal signal in Strecker stent, but not in the Wallstent and Palmaz stent. Strecker stent made of nonferromagnetic tantalum was the best-suited stent for use with contrast enhanced MR angiography. PMID- 9745939 TI - Assessment of commercial compression algorithms, of the lossy DCT and lossless types, applied to diagnostic digital image files. AB - The need for diagnostic image compression of the lossy or irreversible type has been declining due to the rapid increase in commercially available formatted hard disk capacity. It is estimated that the latter has increased about three orders of magnitude in the past 14 years while the size of diagnostic image files has, of course, remained constant. During the same period, despite claims for significantly improved performance by vendors, it seems that only small progress has been made in commercial lossless and lossy compression algorithms. There is still no consensus for lossy compression to a level acceptable for diagnosis. This is mostly considered to be around a ratio of 10:1. However, acceptable compression ratios depend heavily on the type of images processed and may be compared with the 3:1 ratio produced by lossless algorithms. This last value was shown to increase to more than 5.5:1 for gamma-camera images when corrected for the noise content of individual bit planes and for the display capabilities of computer monitors. Therefore, any possible benefits of lossy over lossless compression become questionable when the currently available hard disk capacity and network transmission speed are considered against the inevitable loss of information in the lossy type of compression. PMID- 9745938 TI - Surface matching of multimodality image volumes by a fuzzy elastic registration technique. AB - Multimodality image registration is useful in diagnostic imaging and treatment planning for radiation therapy. In this paper, we present a technique which registers the surfaces of two volumes acquired by different medical imaging modalities. We represent the image volumes in terms of their surface elements known as tiles. We identify the fuzzy variables, assign fuzzy membership functions to them and generate a fuzzy rule database. The fuzzy algorithm reduces the discrepancy between the two set of tiles until the surfaces are matched. In order to study the efficacy of our approach, we severely warp a simulated image and register it with its original. We register CT and MR volumes of humanoid phantom images. Finally, we present the results at the end of the article. PMID- 9745940 TI - Multilevel adaptive process control of acquisition and post-processing of computed radiographic images in picture archiving and communication system environment. AB - Computed radiography (CR) has become a widely used imaging modality replacing the conventional screen/film procedure in diagnostic radiology. After a latent image is captured in a CR imaging plate, there are seven key processes required before a CR image can be reliably archived and displayed in a picture archiving and communication system (PACS) environment. Human error, computational bottlenecks, software bugs, and CR system errors often crash the CR acquisition and post processing computers which results in a delay of transmitting CR images for proper viewing at the workstation. In this paper, we present a control theory and a fault tolerance algorithm, as well as their implementation in the PACS environment to circumvent such problems. The software implementation of the control theory and the algorithm is based on the event-driven, multilevel adaptive processing structure. The automated software has been used to provide real-time monitoring and control of CR image acquisition and post-processing in the intensive care unit module of the PACS operation at the University of California, San Francisco. Results demonstrate that the multilevel adaptive process control structure improves CR post-processing time, increases the reliability of the CR images delivery, minimizes user intervention, and speeds up the previously time-consuming quality assurance procedure. PMID- 9745942 TI - Scrotal cystocele: US and CT findings in two cases. AB - Massive inguinoscrotal herniation of the bladder (i.e. scrotal cystocele) is very uncommon. We describe the ultrasonography and computed tomography features of scrotal cystocele in two patients, and review the radiologic findings of this rare entity reported in the literature. PMID- 9745941 TI - Comparison of MR imaging and CT in discriminating tumor infiltration of bone and bone marrow in the skull base. AB - We compared MR imaging with CT in revealing tumor infiltration of bone and bone marrow in the skull base of 54 patients. MR imaging had no advantages over when tumor involved the anterior compartment. However, precontrast T1-weighted MR images were more efficient than CT in 37.5% of tumors involving the middle compartment and in 54.5% of tumors involving the posterior compartment, respectively. Precontrast T1-weighted images were more accurate than other pulse sequences in revealing bone and bone marrow that were replaced by tumors. PMID- 9745943 TI - Lesions affecting the claustrum. AB - MR imaging examinations of seven patients with a variety of claustral disorders are included in this study. The ages of the patients ranged from 4 to 65 years, and all of them were males. The lesions involving the claustrum comprised cases with asphyxia, Wilson's disease, ischemic white matter disease, thalamic arteriovenous malformation, MELAS syndrome, viral encephalitis and Parkinson's disease. PMID- 9745944 TI - Cerebellar involvement in tuberous sclerosis. AB - Cerebellar involvement in tuberous sclerosis is apparently less than cerebral involvement. In this article, we report CT and MR imaging findings in four tuberous sclerosis patients with apparent cerebellar changes. In one patient with extensive folial calcifications, hypoplasia of the superior vermis, and agenesis of the inferior vermis were associated findings, and clinical correspondence was evident, consisting of a decreased ability to coordinate movements. PMID- 9745945 TI - Partially thrombosed hepatic artery aneurysm mimicking pancreatic head carcinoma. AB - A 43-year-old man with hepatic artery aneurysm which mimicked pancreatic head carcinoma on computed tomography (CT) was studied with B-mode ultrasonography, color doppler ultrasonography, and angiography and the findings are discussed comparing the imaging modalities. PMID- 9745946 TI - Hereditary nephritis (Alport syndrome): MR imaging findings in the brain. AB - Previous clinical studies only described epilepsy and EEG abnormalities in patients with hereditary nephritis (Alport syndrome). In this paper, brain MR imaging findings in a 10-month-old boy with hereditary nephritis are described. These included patchy and nodular lesions in the thalami and basal ganglia, and retarded white matter myelination. The findings suggested an intracranial vasculitis or vasculopathy associated with the syndrome. PMID- 9745947 TI - Caudal regression syndrome: MR appearance. AB - This article reports a case of caudal regression syndrome with hypoplasia of the sacrum, accompanied by an imperforate anus with ano-vestibular fistula. Magnetic resonance images showed a characteristic wedge-shaped cord terminus and the separation of anterior and posterior spinal roots at the level of the cauda equina. PMID- 9745948 TI - Hypoplastic lumbar pedicle in association with conjoined nerve root MRI demonstration. AB - We report a case of aberrant hypoplastic pedicle of the fourth lumbar vertebra and ipsilateral conjoined nerve root. Ipsilateral retroisthmic laminar defect, dysplastic lamina and transverse process, enlargement of neural foramen, hypoplasia of superior and inferior articular facet were present as associated with other neural arch anomalies. The extent of these anomalies was well demonstrated by MR imaging. PMID- 9745949 TI - CT-appearance of intraluminal duodenal diverticulum. The "halo" sign. AB - The barium appearance of intraluminal duodenal diverticulum has been classically described as a "windsock" appearance. However, the CT-scan appearance of this abnormality has not been well documented. A case report of a patient with intraluminal duodenal diverticulum is presented. The authors believe the CT-scan findings in the patient are virtually pathognomonic for this lesion and propose the term "halo" sign be applied to this previously undescribed finding. PMID- 9745950 TI - Identification of mRNA transcripts and immunohistochemical localization of Na/H exchanger isoforms in gerbil inner ear. AB - Recent physiological and pharmacological studies have implicated involvement of the Na/H exchanger (NHE) in regulating inner ear ion homeostasis, but the cellular distribution of this membrane transporter remains unknown. Here reverse transcription and the polymerase chain reaction (RT-PCR) were employed to screen adult gerbil inner ears for mRNA transcripts encoding the four best characterized isoforms of NHE. PCR products spanning selected segments of NHE mRNAs were cloned and sequenced. The putative housekeeping gene NHE-1 was found to be expressed and the 459 bp product shared 98.7% amino acid homology with rat sequence. NHE-2, NHE 3 and NHE-4 cDNA transcripts likewise were detected and the PCR products shared 100, 99.4 and 88.9% amino acid homology, respectively, with their rat counterparts. In addition, the cellular distribution of NHE isoforms 1 and 3 was mapped in the gerbil inner ear by immunostaining with polyclonal antisera against rat antigens. In the cochlea, the antiserum against NHE-1 reacted strongly at the basolateral membrane of strial marginal cells as well as with inner and outer hair cells and spiral ganglion neurons. Less intense staining for NHE-1 was present in subpopulations of fibrocytes in the spiral limbus and in inferior and superior areas of the spiral ligament. In the vestibular system dark and transitional cells expressed abundant NHE-1 as did hair cells and vestibular ganglia neurons. Immunostaining with the antiserum against NHE-3 was limited to the apical surface of marginal cells in the stria vascularis. Based on these data, NHE-1 likely functions primarily to maintain intracellular pH levels in cells where it is found in high abundance. NHE-3, on the other hand, possibly participates in the vectorial transcellular movement of Na+ by strial marginal cells thus helping to maintain the extremely low Na+ level in cochlear endolymph. PMID- 9745951 TI - Evidence for differential regulation of calcium by outer versus inner hair cells: plasma membrane Ca-ATPase gene expression. AB - The expression of mRNA encoding plasma membrane calcium ATPase (PMCA) subunit isoforms (1-4) and splice variants was examined in the adult and developing rat cochlea by PCR and in situ hybridization. High levels of PMCA mRNA expression were observed in the neurons of the spiral ganglion, and in hair cells. Spiral ganglion neurons expressed PMCA 1-3 beginning in embryonic development, reaching high levels shortly after birth, and continuing into adulthood. Inner hair cells expressed PMCA 1 at moderate levels from birth to the time of onset of cochlear function on postnatal day 12, and strongly from then until adulthood. Outer hair cells expressed PMCA 2 at high levels from shortly after birth through adulthood. The data suggest that the calcium clearance requirements of inner and outer hair cells are distinct. PMCA 2 is the isoform with the highest affinity for calmodulin, and has also been associated with high levels of inositol triphosphate. Its presence in outer hair cells suggests that regulation of the enzyme by calmodulin may be particularly important for this hair cell type. It further suggests that inositol phosphate may play a unique role in the outer hair cell. PMID- 9745952 TI - Responses to time-varying stimuli in rat auditory cortex. AB - Responses to frequency modulated (FM) sweeps were recorded in rat primary auditory cortex. Forty-four percent of the cells were direction-selective. For speed selectivity, the majority of the cells preferred faster sweeps. The results suggest that rat auditory cortex may be used for processing communication signals of their predators or for detecting spectral changes in acoustic signals. PMID- 9745953 TI - Mechanisms and effects of transepithelial polarization in the isolated semicircular canal. AB - Previous studies have shown that galvanic stimulation of semicircular canal organs can modulate their afferent discharge. However, it has not been resolved whether this modulation derived from direct stimulation of hair cells, afferent nerve fibers, some combination of the two, or some as yet unknown path. This problem is addressed in the present study. Experiments were designed first to determine the gross current path necessary for the DC current to modulate afferent firing. These led to the conclusion that the current path had to flow between endolymph and perilymph across the neuroepithelium. Next, the various components in this established path were considered: the afferents, the hair cells, between the hair cells, or some combination of the three. These experiments led to the conclusion that the current pathway was across the hair cells causing transmitter release and thus affecting afferent activity. PMID- 9745954 TI - Anatomical evidence for binaural processing in the descending octaval nucleus of the toadfish (Opsanus tau). AB - The connections of a potential auditory circuit were determined in the medulla of the toadfish (Opsanus tau). Fluorescent dextran amines placed in the medial torus semicircularis (mTS) retrogradely filled cells primarily in the dorsal region of the descending octaval nuclei (DON) with contralateral predominance. Fluorescent dextran amines placed in the DON revealed commissural fibers that cross the midline with the internal arcuate tract. The interconnections are consistent with a dorsal-ventral organization of the DON: reciprocal innervation is present for the left and right dorsal zones of the DON and for the left and right ventral zones of the DON. Based on projections to the medial (auditory) TS and the reciprocal connections, the dorsal region of the DON appears to be the major auditory processing site in the medulla and also may be a site for directional, binaural comparisons. The ventral region of the DON may be a site for bilateral vestibular processing. Double-labelling experiments revealed that some of the descending octaval cells projecting to the contralateral DON also project to the mTS. Based on the auditory pathway indicated by this study, future neurophysiological investigations of sensitivity to directional sound stimuli should begin in the dorsal DON of the toadfish. PMID- 9745955 TI - Comparative effects of glycerol and Urografin on cochlear blood flow and serum osmolarity. AB - Glycerol, an osmotic diuretic, has been used for the diagnosis and treatment of endolymphatic hydrops. Hearing improvements in hydropic ears are attributed to its dehydrating effect. In addition to this effect, glycerol also increases cochlear blood flow. Urografin, another hyperosmotic agent used for vasography, is similarly known to increase local blood flow. The present study compared these two hyperosmotic agents, glycerol and Urografin, in their effects on cochlear blood flow and serum osmolarity. Laser Doppler flowmetry on the lateral wall of the cochlea revealed that the increase in cochlear blood flow with a 30-min infusion (0.025 ml/min) of 76% Urografin continued for a longer time than with a 30-min infusion (0.025 ml/min) of 50% (v/v) glycerol. The significant increases appeared at 20 and 30 min after the infusion with the former; 10, 20, 30, 40, 50 and 60 min after the infusion with the latter. Intravenous infusion of these agents also caused elevation in serum osmolarity. This elevation was appreciably greater with Urografin infusion (maximal increase: about 30 mOsm on average) than with glycerol infusion (maximal increase: about 6 mOsm on average), and the former elevation appeared to be longer lasting than the latter. These differences were ascribed to differences between glycerol and Urografin with respect to the creation of an osmotic gradient across the capillary walls of cochlear blood vessels. Since glycerol penetrates the interstitial space and moves into inner ear fluids, the gradient may decline faster. It would be assumed that a higher concentration of the hyperosmotic agent in the capillary blood causes more vasodilatation and lowering of blood viscosity. Alternatively, direct action of these agents on the vascular wall may affect some biological processes, leading to vasodilatation in different degrees and durations with different agents. Hearing improvement with glycerol administration in hydropic ears was also discussed from the perspective of cochlear blood flow. PMID- 9745956 TI - Effects of high sound levels on responses to the vowel "eh" in cat auditory nerve. AB - The vowel "eh" was used to study auditory-nerve responses at high sound levels (60-110 dB). By changing the playback sampling rate of the stimulus, the second formant (F2) frequency was set at best frequency (BF) for fibers with BFs between 1 and 3 kHz. For vowel stimuli, auditory-nerve fibers tend to phase-lock to the formant component nearest the fiber's BF. The responses of fibers with BFs near F2 are captured by the F2 component, meaning that fibers respond as if the stimulus consisted only of the F2 component. These narrowband responses are seen up to levels of 80-100 dB, above which a response to F1 emerges. The F1 response grows, at the expense of the F2 response, and is dominant at the highest levels. The level at which the F1 response appears is BF dependent and is higher at lower BFs. This effect appears to be suppression of the F2 response by F1. At levels near 100 dB, a component 1/component 2 transition is observed. All components of the vowel undergo the transition simultaneously, as judged by the 180 degrees phase inversion that occurs at the C2 transition. Above the C2 threshold, a broadband response to many components of the vowel is observed. These results demonstrate that the neural representation of speech in normal ears is degraded at high sound levels, such as those used in hearing aids. PMID- 9745958 TI - Onset of basilar membrane non-linearity reflected in cubic distortion tone input output functions. AB - The basilar membrane (BM) input output (I/O) function is a non-linear compressive function over much of its operating range. A low level non-compressive region with a break-point or compression threshold between 20 and 40 dB SPL has been identified. To date, no similar compression threshold in cubic distortion tone otoacoustic emission (CDT) data, which would illustrate the dependence of the CDT on BM growth, has been demonstrated. A Taylor series expansion of the outer hair cell gating function yields an amplitude term for 2f1-f2 of p.A1(2).A2, where A1 and A2 are the displacement amplitudes of the BM for two pure tone input stimuli of levels L1 and L2, p a constant. By selectively varying either L1 or L2 with f2/f1 appropriately chosen to reduce suppression effects, the CDT I/O function can be examined for deviation from the power law. In particular, if the amplitude of the CDT were dependent on BM displacement amplitude, then it should be possible by an appropriate choice of parameters to measure compression threshold. We have examined CDT I/O functions for an f2 of 8 kHz in the guinea pig and found them to be consistent with the expected power law. With L1 held constant, L2 varied and f2/f1 = 1.6, a low level region with a slope of one and a compressive region with a slope of 0.14-0.27 corresponding to the analogous regions of the BM I/O function was identified, with a break-point or compression threshold of 22-33 dB SPL. PMID- 9745957 TI - Temporal aspects of the effects of cooling on responses of single auditory nerve fibers. AB - During an investigation of the effects of cochlear cooling on frequency tuning and input/output relations of single auditory nerve fibers in gerbil (Ohlemiller and Siegel (1994) Hear. Res. 80, 174-190), cooling-related changes in post stimulus time histogram (PSTH) shape and phase-locking to tonebursts were characterized in a small sample of neurons. Local cochlear cooling by 5-10 degrees C below normal core temperature did not alter overall PSTH shape, although some evidence was found for a reduction in the time constants of rapid and short term rate adaptation. The relative contributions of rapid and short term response components appeared unaltered. Effects of cooling on phase-locking were assessed by calculating the synchronization index for responses to intense ( > 70 dB SPL) tonebursts at 0.5, 1.0, and 2.0 kHz. Synchronization filter functions exhibited modest reductions in both magnitude and the upper frequency limit of phase-locking. The effects of cooling on the temporal character of responses appear distinct from those of a simple reduction in stimulus intensity. Results are interpreted in terms of cooling-related changes in responses of cochlear hair cells and afferent neurons, and suggest that temperature artifacts are unlikely to underlie reported species differences in PSTH shape and phase locking. PMID- 9745960 TI - Simulation of ILD sensitive neurons in the inferior colliculus of the barn owl. AB - The barn owl (Tyto alba) uses interaural level difference (ILD) as a cue for the localization of sound. The first site of binaural convergence in the pathway that processes ILD is the ventral lateral lemniscus pars posterior (VLVp). Neurons in VLVp receive excitatory input from the contralateral nucleus angularis, and inhibitory input from the contralateral VLVp. Within the lateral shell of the inferior colliculus are ILD sensitive neurons that show maximum spike rate at a specific ILD value, with response falling off sharply on each side. Adolphs has developed a model of such lateral shell neurons based on anatomic and physiological data. In his model, lateral shell neurons receive inhibitory input from VLVp on both sides, and this inhibition, applied against a constant excitatory input, produces the observed two-sided response curves. We simulated, in Matlab 4, Adolphs' model, and obtained supporting results. Our simulation suggests that VLVp provides a repository of simple ILD filters from which higher centers construct more complex filters, including the single-peaked curves observed by Adolphs. The VLVp filters are organized along the inhibitory gradient, with broad filters ventral, sharp filters dorsal. PMID- 9745959 TI - Establishment and characterization of a strial marginal cell line maintaining vectorial electrolyte transport. AB - E6/E7 genes of human papilloma virus type 16 were used to immortalize a primary culture of marginal cells (MC) from gerbils. One of the cloned lines was selected which demonstrated preservation of the main characteristics of the MC, both morphologically and physiologically. Electron microscopic examination showed well developed junctional complexes and apical microvilli which suggested its epithelial origin. Polymerase chain reaction (PCR) demonstrated the incorporation of E6/E7 genes with the genome. Reverse transcription PCR revealed the existence of mRNA of the IsK channel, a unique marker of MC among the inner ear cells, in this clone. Flow cytometric analysis of this cell line's DNA content was diploid. Numerous large domes formed after confluence of the cell monolayer. Electrophysiologic studies displayed evidence of apical K+ and Na+ channels which were blocked by Ba2+ (2 mM) and amiloride (10(-5) M), respectively. Existence of basolateral Na,K-ATPase and Na+/Cl-/K+ cotransporter was shown by blockage by ouabain (10(-3) M) and bumetanide (50 microM), individually. Injection of the cell line to nude mice failed to induce growth of tumors. This cell line was serum-, density- and anchorage-dependent when cultured in plastic dishes. In conclusion, this cell line shows characteristics of well-differentiated MC maintaining the major ionic transport processes, and provides us a good model to study the possible mechanisms and regulating factors of endolymph production. PMID- 9745961 TI - Combined cochleo-saccular and neuroepithelial abnormalities in the Varitint waddler-J (VaJ) mouse. AB - Hearing loss in Varitint-waddler-J (VaJ) mice is of mixed origin with both cochleo-saccular and neuroepithelial components. Both VaJ/VaJ and VaJ/+ mutants show impaired cochlear function, but the homozygotes are more severely affected than heterozygotes. Neither group have any detectable compound action potential. Cochlear microphonics are only seen in half of the heterozygotes, at a reduced amplitude and raised threshold, and are not detected in any homozygotes. Summating potentials (SP) responses are seen in most of the heterozygotes, at high stimulus levels. The only responses in homozygotes were negative SPs seen in half of the mutants at very high sound levels, while the remaining homozygotes showed no responses to sound stimulation. Endocochlear potentials (EP) were often small or absent in both groups of mutants, with the homozygotes being more severely affected. Reduced pigmentation in the stria vascularis appears to be associated with a reduced EP, while a primary defect of the neuroepithelium, detectable by electron microscopy in hair cells of 14 day old mice, dramatically influences evoked potentials. PMID- 9745962 TI - Longitudinal endolymph movements induced by perilymphatic injections. AB - Endolymph movements and endocochlear potential (EP) changes were measured during disturbances of perilymphatic pressure. induced by injecting artificial perilymph into scala tympani (ST) or scala vestibuli (SV) of the guinea pig cochlea. Injections were performed either with or without an outlet made in the opposite perilymphatic scala. Injections into ST without an outlet induced large pressure changes but virtually no endolymph movement or EP change. Injection at the same rate into ST with an outlet in SV produced smaller pressure changes which were accompanied by a basally-directed displacement of endolymph and significant EP changes. The magnitude of endolymph displacements and EP changes varied as a function of injection rate. Injections into SV, either with or without an outlet in ST, produced apically-directed endolymph displacement and EP changes. For the SV injections without an outlet, the cochlear aqueduct and round window are likely to provide an outlet and compliance, permitting flow along the perilymphatic scalae to occur even when no ST outlet was provided. We conclude that endolymph movements are not dependent on the absolute pressure of the perilymph, but instead occur when small, sustained pressure gradients are present across the cochlear partition, corresponding to times when perilymph flow is induced. This study demonstrates that in the normal. sealed cochlea, endolymph and EP are insensitive to fluid injections into ST, but are sensitive to fluid injections into SV. Endolymph movements are therefore unlikely to be generated by cerebrospinal fluid pressure fluctuations (such as those produced by respiration, posture changes, coughing, sneezing, etc) which are transmitted to ST by the cochlear aqueduct. PMID- 9745963 TI - Postnatal vascular development in the lateral wall of the cochlear duct of gerbils: quantitative analysis by electron microscopy and confocal laser microscopy. AB - The development of the capillary network in the stria vascularis and in the underlying spiral ligament of gerbils was systematically and quantitatively investigated by conventional electron microscopy and confocal laser microscopy in association with vascular labeling with fluorescent gelatin. The developmental changes of capillaries in the lateral wall were observed as the following series of events. (i) At 0 days after birth (DAB) capillaries already existed in the spiral ligament as a network. (ii) At 3-9 DAB the capillary network developed into two layers starting from the scala vestibuli side to the scala tympani side; one layer was located in the stria and the other in the spiral ligament. (iii) At 9 DAB capillaries in the stria became separated from the spiral ligament, and the capillary network consisting of a two-layered structure was complete. (iv) Total capillary length and capillary density in the lateral wall increased until 9 DAB and leveled off thereafter, but changes in the relative position of capillaries in the stria toward the luminal surface of marginal cells continued until 31 DAB. On the basis of the above observations, we propose two possible mechanisms underlying the vascular development in the lateral wall: (i) the formation of new vasculature (angiogenesis), and (ii) changes in the position of cellular components relative to capillaries in association with the differentiation and maturation of marginal cells and intermediate cells. PMID- 9745964 TI - Divalent cations inhibit IsK/KvLQT1 channels in excised membrane patches of strial marginal cells. AB - The IsK/KvLQT1 K+ channel in the apical membrane of strial marginal cells and vestibular dark cells is an essential ion transport pathway for the secretion of K+ into the endolymph of the inner ear. Study of this control point has been impeded by rundown of channel activity upon excision into commonly used cytosolic solutions. This paper describes conditions under which patches of apical membrane of strial marginal cells and vestibular dark cells from gerbil containing this channel can be excised, retaining its characteristic voltage dependence, kinetic properties, ion permeability sequence and pharmacological sensitivity, similar to those found during on-cell and perforated-patch whole cell recordings (Shen et al., Audit. Neurosci. 3 (1997) 215-230). Those excised-patch conditions include removal of Mg2+ from the cytosolic solution and use of a K+-rich pipette electrolyte. The inhibition of channel activity by Mg2+ was found to be a general feature of divalent cations; the channel was also inhibited by Ca2+, Ba2+ and Sr2+. The concentrations causing 50% inhibition of IsK/KvLQT1 channel current were 7 x 10(-5) M, 6 x 10(-6) M, 3 x 10(-4) M and 7 x 10(-5) M, respectively. It was also found that a chemical cross-linking agent, 3,3'-dithio bis(sulfosuccinimidyl propionate) (DTSSP), which was previously shown to persistently activate IsK/KvLQTI channels expressed in Xenopus oocytes, maintained in excised patches channel activity which retained voltage dependence and pharmacological sensitivity. These data demonstrate that (1) the channel complex is inhibited by Ca2+, Mg2+ and other divalent cations, (2) the activation by Ca2+ observed previously in whole-cell preparations was due to action via other cellular pathways. These findings must be taken into account when considering the action of receptors which alter the cytosolic Ca2+ activity. PMID- 9745966 TI - General characteristics and suppression tuning properties of the distortion product otoacoustic emission 2f1-f2 in the barn owl. AB - The distortion-product otoacoustic emission (DPOAE) 2f1-f2 was measured in the ear canal of the barn owl. DPOAE were elicited by primary tones in 11 frequency regions from 1 to 9 kHz. The highest DPOAE output levels and best thresholds were found for f1 frequencies of 4 to 7 kHz and additionally at the lowest f1 frequency investigated. In some cases, the DPOAE sound pressures were only 37 dB below the primary-tone levels (PTL). The optimal primary-tone frequency ratios ranged from 1.05 to 1.45 and varied strongly among the different frequency regions investigated. The largest optimal ratios were measured in the middle frequency range for f1. At lower and higher f1, the optimal ratios decreased. DPOAE levels could be suppressed in a frequency-selective way by adding a third tone. As in other non-mammals, the best suppressive frequencies were near f1, suggesting DPOAE generation near the frequency place of this primary tone. This is in contrast to what is known for mammalian species, where the DPOAE is thought to be generated near f2. To obtain 6 dB of suppression of the DPOAE level, suppressor-tone levels ranging from 13 dB below to 4 dB above the primary-tone level were necessary. The Q10dB-values of suppression tuning curves increased as a function of frequency up to a value of 15.8. This tendency resembled the increase in frequency selectivity of auditory nerve fibers in this species. PMID- 9745965 TI - Outward rectifying potassium currents are the dominant voltage activated currents present in Deiters' cells. AB - Supporting cells in the cochlea are thought to maintain the homeostasis of the organ of Corti and contribute to the electrical and micromechanical environment of the hair cells. Of the different types of supporting cells, Deiters' cells form a structure that holds the outer hair cells (OHCs) at their base and apex. This structure may play an important role in modifying cochlear mechanics by influencing the force produced by sound induced motion of the OHCs which in turn may be modulated by ATP acting on ligand gated cation channels on the Deiters' cells. Also, a glia-like role of buffering external K+ concentration for the Deiters' cells has been suggested. We studied Deiters' cells' electrical properties and ion conductances using the whole cell variant of the patch clamp technique since they must play an important role in the function of these cells. It was found that isolated Deiters' cells possess a large voltage activated, outwardly rectifying K+ selective conductance. Voltage activated Ca2+ currents and non-selective currents were not detected and voltage activated inward currents were very small. The outward K+ currents were found to be dependent on voltage but not on Ca2+ for their activation. Nimodipine and 4-aminopyridine (4 AP) were shown to interact directly with the K+ channels in a voltage dependent manner. It is suggested that the K+ selective channels in Deiters' cells may be similar to the Kv1.5 type channel. However, based on the voltage dependence of the channels that was described by double Boltzmann equation and on the alteration of that dependence by 4-AP, it is possible that more than one type of K+ selective channel exists. PMID- 9745967 TI - The use of high stimulus rate auditory brainstem responses in the estimation of hearing threshold. AB - This normative study investigates the efficiency of using the maximum length sequence (MLS) technique applied to auditory brainstem evoked response (ABR) testing to estimate hearing thresholds. Using a commercially available system, ABRs were recorded in sixteen subjects at two conventional rates--9.1 and 33.3 clicks/s--and six MLS rates between 88.8 and 1000 clicks/s. Each subject was tested at five stimulus levels from 60 down to 10 dBnHL. The wave JV amplitude input-output (I/O) functions, relative signal to noise ratio (SNR) and speed of test were calculated for all conditions. The JV amplitude and detectability decrease as the stimulus rate increases and level decreases. The latency of JV increases as the stimulus rate increases and the intensity decreases. While the slope of the amplitude I/O function was maximal at 200 clicks/s, at 300 clicks/s it was comparable with that obtained at conventional rates. At higher rates, the slope of the I/O function decreases. When compared with the conventional recording rate of 33.3 clicks/s there is a small improvement in SNR for MLS rates between 200 and 600 clicks/s at levels above 30 dBnHL. The calculated speed improvement at 300 clicks/s is a factor between 1.4 to 1.6 at a screening level of 30-40 dBnHL. It is felt therefore that there may be a small advantage to using MLS in screening and that the optimal rate for this lies at around 200 to 300 clicks/s. However even at these rates, as a consequence of the adaptation of the response with both rate and level, the improvement in SNR or speed of test would be modest when estimating threshold. PMID- 9745968 TI - Economic costs of urinary incontinence. PMID- 9745969 TI - Nocturnal enuresis: pathogenesis and treatment. AB - A new concept is put forward to explain the act of micturition and urinary continence. This depends mainly on, the presence of an intact, sound and strong internal sphincter, and, an acquired behavior gained by learning in early childhood how to maintain a high alpha-sympathetic tone, thus keeping the internal sphincter closed all the time. On desire, and at the appropriate time and place, this acquired high alpha-sympathetic tone is inhibited, thereby opening the internal sphincter and allowing voiding to occur. The aim of the study was to test the effects of giving an alpha-sympathomimetic drug, ephedrine hydrochloride to control nocturnal enuresis. Three hundred patients suffering from enuresis were evaluated by a systematic history, physical examination, urinalysis and urodynamic studies. They were divided into a study group of 150 and a control group of 150. The study group were given ephedrine hydrochloride tablets after lunch and before bedtime for 2 months. The control group were given one of the following: imipramine hydrochloride, desmoporessin, ritaline, cetiprin, or behavioral therapy. Immediate continence was gained by 86% of the study patients; 6.7% had slower gain of continence. In the control group there was a short-term success rate of 30%-70% while on treatment, and a high relapse rate of more than 90% after stopping the treatment. It was concluded that giving ephedrine hydrochloride will improve internal sphincter tone, thus preventing uncontrolled urination. PMID- 9745970 TI - Postoperative urinary tract infections (UTIs) following single-dose intraoperative antibiotic prophylaxis in colposuspension patients. AB - The records of 196 women who underwent colposuspension for genuine stress incontinence at the Leicester General Hospital, England, between June 1991 and May 1996 were reviewed for evidence of urinary tract infection (UTI). Variables analyzed include age, type of antibiotic, timing of a positive culture, organism(s) responsible and antibiotic sensitivity. Forty-six patients (23.47%) developed urinary infection; of these, 42 had received single-dose antibiotic prophylaxis with suprapubic catheterization. Thirty-two (76%) of those who developed UTI received augmentin (amoxycillin and clavulanic acid), whereas 10 (24%) were given cefuroxime and metronidazole. Positive cultures were obtained between postoperative days 3 and 28, with a mean of 9.6 days, and 81% occurred after the 7th day. Coliform organisms were responsible for nearly 70% of the infections. UTI is still common after colposuspension, despite single-dose antibiotic prophylaxis. Further studies looking at longer or alternative courses of antibiotics or clean intermittent self-catheterization are essential to establish the best way of curbing UTI in urogynecology patients. PMID- 9745971 TI - Non-evidence of estrogen receptors in the rectal mucosa. AB - The aim of the study was to investigate whether estrogen receptors are present in the rectal mucosa of premenopausal women compared to postmenopausal women and men. Thirty biopsies obtained from the rectal mucosa at colonoscopy, performed to investigate inflammatory bowel disease in 23 patients and neoplasia in 7, were examined by the avidin-biotin-peroxidase immunohistochemical technique for the presence of estrogen receptors. The study group (n = 10) were non-pregnant premenopausal women and the control group (n = 20) were postmenopausal women (n = 10) and men (n = 10). None of the subjects had fecal incontinence or was taking medication with hormones. In no case did the primary lesion involve the specimen used for laboratory analysis. All samples showed negative immunostaining for estrogen receptors. It was concluded that in continent women and men, a direct estrogenic effect on the rectal mucosa seems unlikely. PMID- 9745972 TI - The influence of obesity, constitution and physical work on the phenomenon of urinary incontinence in women. AB - Urinary incontinence in women is a common and complex problem which can be defined and classified as stress, urge and mixed incontinence. Three of the eight most common risk factors are obesity, constitution and physical work, in addition to age, length of menstrual cycle, number of pregnancies, education and level of health awareness. Women with the diagnosis of urinary incontinence were invited to respond to questionnaires on a voluntary basis. The three factors found to be associated with urinary incontinence are increased body weight, strong osteomuscular structure and hard physical work. These indicate that the work of a health team must take a holistic approach to women even before the phenomenon of urinary incontinence occurs. PMID- 9745973 TI - Randomized double-blind placebo-controlled multicenter evaluation of efficacy and dose finding of midodrine hydrochloride in women with mild to moderate stress urinary incontinence: a phase II study. AB - Midodrine is a potent and selective alpha1-receptor agonist and its potential to increase urethral closure pressure could be useful in the treatment of female stress incontinence. The aim of this randomized double-blind placebo-controlled multicenter study was to evaluate the efficacy and safety of midodrine for the treatment of stress urinary incontinence. The primary criterion of efficacy was the maximum urethral closure pressure at rest. Voiding diaries, symptom and incontinence questionnaires and patient/investigator global assessment were also used to evaluate its efficacy. After 4 weeks of treatment no significant changes in MUCP were found. The global assessment by the patient and investigator did indicate that patients on active treatment had a more positive assessment than the placebo group. In conclusion, midodrine did not cause significant improvements in urodynamic parameters, but there were subjective improvements in some of the patients in the treated groups. Furthermore midodrine was well tolerated. PMID- 9745974 TI - Efficacy of biofeedback in the treatment of urinary stress incontinence. AB - Thirty-seven women with stress incontinence were given biofeedback instruction on how to perform pelvic floor exercises correctly. After 3 months with home exercises 31 patients performed a new standardized pad-weighing test: 39% were objectively cured and 42% improved. After a mean of 2 years 15 patients were evaluated with another pad-weighing test: 27% were now objectively cured and 47% improved. A questionnaire showed that 78% had an exact knowledge about the location of the pelvic floor muscles and 47% were satisfied with their present situation, but only 58% performed daily exercises. PMID- 9745975 TI - The pubourethral ligaments--an anatomical and histological study in the live patient. AB - The aim of the study was to analyze the structure, relations and insertions of the pubourethral ligament in the living female. Thirty-five women, mean age 44 years, were studied. The intravaginal slingplasty (IVS) procedure, as performed via two paraurethral incisions, allowed immediate access to the structures in this area, the urethra, vaginal hammock, pubourethral ligaments and anterior portion of the pubococcygeus muscle. Histological biopsies were performed from the structures identified as ligaments. The pubourethral ligament descends like a fan from the lower part of the pubic bone. It consists of vaginal and urethral parts, joined together by thin fibrous threads, giving the appearance of a continuous sheet of amorphous connective tissue. Each part generally varies between 5 and 7 mm in width and 3-4 mm in thickness. The urethral part is approximately 2 cm long and inserts into the midpart of the urethra. The vaginal part is approximately 3-4 cm long. It inserts into the vaginal hammock posterolaterally, approximately 1 cm short of the bladder neck. Histologically the ligaments consist of smooth muscle, elastin, collagen, nerves and, blood vessels. The dissections confirm that the pubourethral ligaments are strong finite structures. Allowing for differences between cadavers and live patients, relationships and insertions are much as described by Robert Zacharin. PMID- 9745976 TI - Urinary incontinence in women, without manifest injury to the bladder. 1914. PMID- 9745977 TI - Fascia lata sling cystourethropexy for the management of female urinary incontinence. AB - Pubovaginal sling cystourethropexy has rapidly become one of the primary surgical treatment options for women with urinary incontinence. The procedure has evolved over time with regard to clinical indications, patient selection criteria and surgical techniques. This article reviews the historical development of pubovaginal sling cystourethropexy, including recent technical advances. The selection of graft materials is considered and the utility of fascia lata emphasized. Clinical results and potential complications of the procedure are also reviewed. PMID- 9745978 TI - Interstitial cystitis. AB - Interstitial cystitis (IC) is a multifactorial syndrome with symptoms of pelvic or perineal pain, urinary frequency and urgency. The etiologies are unknown, but several theories have been proposed. Diagnosis is often delayed because most of the conventional evaluation is normal. Pelvic examination is normal except for bladder tenderness. Urodynamics are normal except for increased bladder sensitivity and low capacity. Urinalysis, urine culture and office cystoscopy are also normal. The diagnostic test is cystoscopy under anesthesia with bladder distension. Small submucosal hemorrhages (glomerulations) or ulcers appear after distension. Many empiric treatments have been proposed for IC. None is universally effective, and so treatments are tried sequentially until good symptom relief is achieved. Bladder distension gives excellent (but transient) relief in some patients, especially those with severe bladder inflammation (who also tend to be older). A variety of oral, intravesical and adjunctive treatments are also described. PMID- 9745979 TI - Inadvertent ureteral catheterizaion with a microtip catheter at cystometry. AB - Two patients in whom the right ureter was inadvertently catheterized at water cystometry are described. Accidental ureteral catheterization and filling was followed by colicky pain in the right flank and by an abrupt increase in the recorded pressure, up to 148 cm H2O. The pain disappeared and the intravesical pressure returned to baseline after the microtip catheter was withdrawn. PMID- 9745980 TI - Maternal-fetal transfer of melatonin in pregnant women near term. AB - Our objective was to evaluate the maternal-fetal transfer of melatonin in pregnant women. Serum melatonin concentration was measured by high-performance liquid chromatography with electrochemical detection in a maternal vein and in the umbilical artery and umbilical vein at the time of birth. Blood samples were obtained from 12 women who had spontaneously delivered vaginally at night. A single oral dose of melatonin was administered to each of 33 patients who underwent a cesarean section, and, blood samples were taken at 1, 2, 3, or 4 hr after the administration of melatonin at delivery. Cesarean section was performed between 1300 and 1500 hr. The mean melatonin concentrations of melatonin in maternal peripheral venous blood and umbilical arterial and umbilical venous blood did not differ significantly, and positive correlations in the serum levels of melatonin were observed between the three sources of blood. The oral administration of 3 mg of melatonin to pregnant women led to marked increases in the serum levels of melatonin, with maximum levels observed 2 hr (21.84 +/- 2.09 ng/ml) after drug administration. Changes in serum levels of melatonin in the umbilical vein and artery resembled those found in the maternal vein. Serum melatonin concentrations did not differ significantly between the maternal vein and the umbilical veins. Serum levels of melatonin in the umbilical vein after the administration of melatonin were significantly and closely correlated with those in the maternal vein (r = 0.924, P < 0.001). These results suggest that, in humans, melatonin is transferred from the maternal to the fetal circulation both easily and rapidly. A potential for the therapeutic use of melatonin as an antioxidant exists in the patients with preeclampsia. PMID- 9745981 TI - Melatonin directly suppresses steroid production by preovulatory follicles in the cyclic hamster. AB - The purpose of these studies was to investigate the effects of melatonin on the production of steroids (progesterone, testosterone, and estradiol) and cAMP by preovulatory follicles and to examine changes in melatonin concentrations in the ovary during the estrous cycle. Adult cyclic hamsters were used in this study. Melatonin concentrations in the ovary, pineal gland, and serum were measured at mid-light and mid-dark during the estrous cycle. Effects of melatonin on steroidogenesis by preovulatory follicles, thecae, and granulosa cells were examined, and its effect on cAMP production by preovulatory follicles was also investigated. Melatonin (0.1-10 ng/ml) had no effect on steroid production in the absence of hCG, but melatonin decreased progesterone and estradiol production by preovulatory follicles in a dose-dependent and time-dependent manner in the presence of hCG (100 mIU/ml). The target of melatonin was thecae but not granulosa cells, and melatonin significantly reduced cAMP production by preovulatory follicles. Melatonin concentrations in the ovary showed a similar phasic variation with high levels during mid-dark and low during mid-light, as in the pineal gland and serum. These results show that the ovarian melatonin levels also exhibit a circadian rhythm and suggest that the high melatonin milieu in the ovary may induce gonadal regression in the cyclic hamster. PMID- 9745982 TI - Time-dependent effects of melatonin on immune measurements in male Syrian hamsters. AB - Adult, male Syrian hamsters received daily subcutaneous melatonin (25 microg) injections or vehicle injections at 08:00 or 17:00 hr for 11 weeks. Body weights were measured weekly throughout the experiment and testes weights, spleen weights, and serum was collected at the end of the experiment. The spleens were sectioned and immunocytochemically analyzed for immunoglobulin G and serum levels of interferon-gamma, interleukin-2, and interleukin-4 were determined in heterologous mouse assays. Melatonin injections at 17:00 hr, but not at 08:00 hr, increased body weights, decreased testes weights and serum testosterone levels, and had no effect on immunoglobulin G content in the spleen. Likewise, melatonin injections at 17:00 hr, but not at 08:00 hr, increased serum interferon-gamma levels, had no effect on interleukin-2 levels, and appeared to increase interleukin-4 levels. Since melatonin injections at 08:00 hr were ineffective in altering immune measurements and correlations between reproductive measures and immune measures were high, the most parsimonious explanation for these results is that melatonin injections at 17:00 hr depressed reproductive hormone levels and these depressed levels altered immune measures. PMID- 9745983 TI - Neuroendocrine, immunohistochemical, and ultrastructural study of pineal region tumors. AB - Thirteen patients with tumors in the pineal region were submitted to pre- and post-operative blood sampling (08:00, 14:00, 20:00, and 02:00 hr) for three or four consecutive days. A single cerebrospinal fluid (CSF) sample was collected at surgery, and melatonin levels determined. In all patients, serum and CSF beta subunit of human chorionic gonadotrophin (betaHCG), carcino embryonic antigen (CEA), and alpha-fetoprotein (AFP) levels were measured. Histology revealed four pineocytomas, one pineoblastoma, four germinomas, one immature teratoma, one pilocytic astrocytoma, one lymphoma, and one meningioma. Serum and CSF levels of serological biomarkers were normal, except for one of the germinoma cases. In most patients, alteration either in the circadian rhythm or in the melatonin concentration was observed before surgery. In benign neoplasms the circadian rhythm was conserved. In pineoblastoma, lymphoma, and three out of four germinomas, melatonin concentrations were undetectable. In one case of germinoma, melatonin levels were high, with the circadian rhythm being abolished. According to conventional histology, all germinomas were similar. Therefore, in a rare case of pineal germinoma with high melatonin levels, the tissue was subjected to an in depth investigation (immunohistochemical and ultrastructural) in order to determine the pathology and the possible differences from the other typical germinomas. Results were compared to those provided from other pineal neoplasms. Electron microscopy examination detected the presence of clusters of intermediate filaments and numerous electrondense granules only in the case of a germinoma producing melatonin. PMID- 9745984 TI - Twenty-four hour rhythm of melatonin in patients with a history of pineal and/or hypothalamo-neurohypophyseal germinoma. AB - Melatonin deficiency after a pinealectomy has been investigated in animals; however, in humans, this status can be assessed solely by investigating patients with a tumor originating in the pineal gland. This study analyzes secretion of melatonin and pituitary hormones in 14 patients with germinoma originating in the pineal or the hypothalamic-neurohypophyseal region. Thirteen patients had been successfully treated prior to this study. One patient was included in this study before the initiation of treatments. Plasma sampling was performed every 2 hr for 24 hr and melatonin concentrations were measured by radioimmunoassay. Melatonin secretion was nearly absent in the patients with pineal germinoma regardless of treatment option, even in the patient who had been untreated. In contrast, melatonin secretion and its circadian rhythms were not affected in patients with a hypothalamo-neurohypophyseal germinoma. The circadian rhythms of growth hormone and adrenocorticotropic hormone were not dysregulated in patients with the melatonin deficiency. We conclude that germinoma cells originating the pineal gland impair the production of melatonin by pineocytes and consequently induce a permanent melatonin deficiency in those patients. Since melatonin exerts multiple physiological functions, once a clinical concept of "melatonin deficiency syndrome" is established, melatonin replacement therapy could be investigated in patients who have a pineal germinoma or who have undergone a neurosurgical pinealectomy. PMID- 9745985 TI - Urinary 6-sulfatoxymelatonin profiles in male Djungarian hamsters (Phodopus sungorus) responding and not responding to short-day photoperiods: possible role of elevated daytime levels. AB - The lack of endocrine and physiological responses of some Djungarian hamsters (Phodopus sungorus) to the transition from long to short photoperiods (L:D 16:8 - > L:D 8:16) has been known for a long time but is not yet understood. We investigated the role of melatonin synthesis in this context because melatonin, as part of the circadian system, may play a role in non-responsiveness. In ten responding and ten non-responding male hamsters, the urinary 24 hr 6 sulfatoxymelatonin (aMT6s) profiles under L:D 8:16 and L:D 16:8 were measured. Both short day responding and non-responding hamsters showed diurnal aMT6s excretion rhythms. Whereas responders reacted to the transition L:D 16:8 --> L:D 8:16 with a marked elevation of aMT6s excretion, in non-responders no adjustment of the melatonin rhythm to the change of the photoperiod was seen. Furthermore, under L:D 16:8 the daytime levels of aMT6s were significantly (P<0.001) lower in responders compared to non-responders whereas under L:D 8:16 these levels were higher (P<0.01). It is speculated that high daytime levels of aMT6s under long day photoperiods in non-responders result in down-regulation of melatonin receptors of the nucleus suprachiasmaticus, the pacemaker for the pineal gland, leading to a lack of response to the transition to short-day photoperiods. PMID- 9745986 TI - Effects of two melatonin analogues, S-20098 and S-20928, on melatonin receptors in the pars tuberalis of the rat. AB - By using quantitative autoradiography, we studied the effects of two drugs related to melatonin on the 2-(125)I-melatonin binding in the pars tuberalis (PT) of rats. The drugs tested were two naphthalenic analogues of melatonin, S-20098 (N-[2-(7-methoxy-1-naphthyl) ethyl] acetamide), an agonist, and S-20928 (N-[2-(1 naphthyl) ethyl] cyclobutyl carboxamide), a putative antagonist. Melatonin (s.c. and i.p.), S-20098 (s.c.), and S-20928 (i.p.) were injected 4 hr before sacrifice. Acute administration of both melatonin and S-20098 decreased melatonin receptor density. In contrast, the putative antagonist S-20928, at a low dose (1 mg/kg), was ineffective on melatonin receptors. It neither affected the 2-(125)I melatonin specific binding observed in the control group nor did it prevent the decrease in binding induced by melatonin when injected 5 min before the hormone. At a high dose (10 mg/kg), S-20928 totally blocked the effect of melatonin on melatonin receptor density and induced a decrease in binding capacity as melatonin did when injected alone. These results indicate that in the rat pars tuberalis, the melatonin agonist, S-20098, is able to down-regulate melatonin receptors, whereas S-20928 seems to behave as a partial agonist. PMID- 9745987 TI - Melatonin effects on sleep, mood, and cognition in elderly with mild cognitive impairment. AB - The effects of immediate-release melatonin on circadian rest-activity profiles, cognition, and mood were investigated in ten elderly individuals with self reported sleep-wake disturbances. Melatonin (6 mg), administered 2 hr before habitual bedtime, enhanced the rest-activity rhythm and improved sleep quality as observed in a reduction in sleep onset latency and in the number of transitions from sleep to wakefulness. However, total sleep time was not significantly increased nor was wake within sleep significantly reduced. The ability to remember previously learned items improved along with a significant reduction in depressed moods. No side effects or contraindications were reported by any of our participants during the 10 day trials. These data suggest that melatonin can safely improve some aspects of sleep, memory, and mood in the elderly in short term use. PMID- 9745988 TI - Ischemia/reperfusion-induced arrhythmias in the isolated rat heart: prevention by melatonin. AB - Cardiac arrhythmias during ischemia/reperfusion are believed to be related to free radicals generated in the heart especially during the period of reperfusion. Since melatonin functions as a free radical scavenger and antioxidant, the ability of this molecule to influence cardiac arrhythmias was investigated. The pineal secretory product, melatonin, reduced the incidence and severity of arrhythmias induced by ischemia/reperfusion due to ligation of the anterior descending coronary artery in the isolated rat heart. Melatonin was either infused during both the ischemia and reperfusion periods or only late in the ischemia period and throughout reperfusion. The percentage of hearts that developed cardiac arrhythmias during reperfusion as indicated by the incidence of premature ventricular contraction (PVC) and ventricular fibrillation (VF) were recorded. Melatonin either infused during both the ischemia and reperfusion periods or during essentially the period of reperfusion greatly reduced PVC and VF due to occlusion and reopening the anterior descending coronary artery. Presumably melatonin's beneficial effect in reducing cardiac arrhythmias was due in part to its free radical scavenging activity, which is greatly assisted by the rapidity with which it is taken up into cells. Previous studies have shown that vitamin C is effective in reducing the severity of cardiac arrhythmias induced by ischemia/reperfusion; thus, we also compared the efficacy of melatonin with this well-known antioxidant. Melatonin was more potent than vitamin C in protecting against arrhythmias induced by ischemia/reperfusion. Besides melatonin's function as a broad spectrum free radical scavenger, melatonin may have also reduced cardiac arrhythmias due to its regulation of intracellular calcium levels, i.e., by preventing calcium overloading, or due to its ability to suppress sympathetic nerve function and reduce adrenergic receptor function in the myocardium. Additional studies into the mechanisms of melatonin's action in reducing cardiac arrhythmias due to ischemia/reperfusion or other causes are warranted because of the possible application of this information to humans with heart disease. PMID- 9745989 TI - Clinical benefits and problems in recent automated peritoneal dialysis treatment. PMID- 9745990 TI - Hemofiltration in children. PMID- 9745991 TI - Clues for understanding the pathogenesis of left ventricular hypertrophy in chronic uremia. PMID- 9745992 TI - Effects of dialysis membranes on the kinetics of tumor necrosis factor-alpha production by peripheral mononuclear cells in chronic hemodialysis patients. AB - To evaluate the biocompatibility of dialysis membranes, blood samples were collected from 10 hemodialysis patients immediately before dialysis and peripheral blood mononuclear cells were isolated. The 3.0 x 10(5) cells/ml were then passed 30 times through modules made of a polyethylene glycol-grafted cellulose membrane, a polyacrylonitrile membrane, and a polysulfone membrane. Expression of messenger RNA for tumor necrosi factor-alpha (TNF-alpha) was determined. Cells were also cultured for 2 h with and without lipopolysaccharide and TNF-alpha levels in the supernatant were measured. TNF-alpha messenger RNA expression was significantly higher immediately after passage through the polyacrylonitrile membrane compared with the other membranes. Cells cultured without lipopolysaccharide, produced significantly less TNF-alpha after passage through the polysulfone membrane, while lipopolysaccharide significantly increased TNF-alpha production by cells passed through the polyacrylonitrile membrane. These results suggest that biocompatibility differs even among dialysis membranes believed to cause no complement activation. PMID- 9745993 TI - Levels of sustainable aerobic workload in dialysis patients. AB - The aerobic performance of a heterogeneous group of 89 ambulatory medically stable patients on chronic hemodialysis was studied to define individual levels of the most acceptable metabolic workload. The patients performed a step test protocol (3 steps) with a cycle ergometer. Each step (25 Watt) lasted 6 min. Heart rate (HR), oxygen consumption (VO2), ventilation (VE), respiratory exchange ratio (RER), blood pressure (BP) and subjective ratings of dyspnea (CRd) and fatigue (CRf) levels were monitored throughout the test. The test was continued to exhaustion or to values of systolic arterial blood pressure (SABP) >240 mmHg, heart rate (HR) > or = 85% max, or ST changes in ECG. In eleven patients (12.5% of the whole group) the test was interrupted within the first two minutes of exercise. In the remaining 78 patients, the maximum workload sustained for at least 3 min (MSW) was 25 Watts for 43 (48.5%), 50 Watts for 27 (30%), and 75 Watts for 8 (9%) patients. Performance was affected in a statistically significant manner by the subjects' anagraphic age, but not by their dialytic age, hemoglobin (Hb) level or weight. Individual levels of tolerable workload were estimated for 60% of the group from the stability of physiological variables during 3 min, and from subjective ratings at a "moderate" level. This level corresponded to an average of 3.5+/-0.9 METs, at 60% of the HRmax, with a mean BP of 167+/-21/98+/-14 mmHg. This could become a safe starting point for a program of physical retraining. PMID- 9745994 TI - Evidence of profiled hemodialysis efficacy in the treatment of intradialytic hypotension. AB - In the last 10 years the percentage of dialysis patients suffering from clinical intradialytic intolerance has greatly increased. Profiled hemodialysis (PHD) is a new technical approach, alternative to standard hemodialysis (SHD) for the treatment of intradialytic symptomatic hypotension. It is based on intradialytic modulation of the dialysate sodium concentration, using a dialysate sodium concentration profile elaborated by a new mathematical kinetic model. The aim of PHD is to reduce the intradialytic blood volume decrease, thanks to a dialysate sodium profile, which allows a reduction in the plasma osmolarity decrease, thereby boosting intravascular fluid refilling. This work aims at clinically validating the PHD technique, by testing its ability against SHD, to maintain a more stable intradialytic blood volume; this evaluation was supported by monitoring some hemodynamic parameters. Twelve dialysis patients on SHD treatment were selected because of their intradialytic symptomatic hypotension. Twelve SHD (one per patient) and 12 PHD sessions (one per patient) were performed to achieve the same sodium mass removal and body weight decrease on both PHD and SHD. During these sessions we monitored the blood volume variation % by the crit-line (a non invasive blood volume monitoring device), the mean blood pressure and heart rate directly and, finally, the stroke volume and cardiac output indirectly by bidimensional doppler-echocardiography. Comparison of the results obtained with the two techniques shows PHD to achieve a significantly more stable blood volume, blood pressure and cardiovascular function than SHD, in particular during the second and the third hour of the dialysis session. PMID- 9745995 TI - Fever in dialysis patients with recently rejected renal allografts. AB - INTRODUCTION: Fever of unknown origin is a complex problem in dialysis patients with recently rejected renal allografts, due to the contribution of the newly withheld immunosuppressive agents to the immunosuppression of uremia, resulting in an atypical presentation of infections, a main cause of fever in these cases. MATERIALS AND METHODS: Two dialysis patients with recently rejected renal allografts who were hospitalized because of fever of unknown origin are reported. Biochemical, bacteriological and imaging studies were performed for specific diagnosis. RESULTS: Extensive laboratory investigations failed to yield any diagnosis and allograft nephrectomy was performed in one patient, with a probable diagnosis of inflammation of the allograft, which resulted in no improvement. Eventually, both patients were found to have adrenal insufficiency responsible for the fever, which improved after steroid replacement. CONCLUSIONS: Adrenal insufficiency should be suspected in all dialysis patients presenting with fever and atypical symptoms, but only after other potential causes are eliminated; since steroid administration may normalize fever regardless of the etiology, it may mask the signs and symptoms and delay the treatment of other (if any) underlying disorder(s). PMID- 9745996 TI - Biocompatibility evaluation of polyamide hemofiltration. AB - INTRODUCTION: Postdilution hemofiltration with a polyamide membrane is a renal replacement technique widely used, but very little information is available regarding the biocompatibility of this treatment. In this paper we report the results of an acute study of the biocompatibility of polyamide hemofiltration. PATIENTS AND METHODS: Complement activation such as C3a and C5a Des Arg (RIA), granulocyte degranulation like alpha 1 elastase intradialytic increase (ELISA) and the expression of high affinity membrane receptors for IL-2 (anti-TAC) were determined. Beta 2-microglobulin (RIA) intradialytic decrease, as well as its convective removal, was evaluated. The nature of protein layer adsorbed onto the polyamide membrane, at the end of the dialytic session was investigated with a new immunohistochemical technique. Cell-associated cytokine concentration (like IL-1 beta and IL-1Ra - ELISA) was determined on mononuclear cell lysates. RESULTS: A low degree of complement activation was detected with the polyamide membrane when data were adjusted for hemoconcentration and for 1 m2 of membrane surface area. An important convective removal not only of Beta 2-microglobulin (258+/-20 mg/session), but also of the activated anaphylatoxins (225+/-76 ng/ml for C3a and 22.5+/-4 ng/ml for C5a) was revealed. A marked deposition of all coagulation factors with no detectable amount of immunoglobulins and complement factors was revealed on the polyamide membrane at the end of the dialytic session. No intradialytic (for IL-1beta) (from 14. 1+/-3.0 to 13.5+/-2.9 pg/2.5 x 10(6) cell) and interdialytic (for IL-1Ra) (from 4572+/-1076 to 5408+/-615 pg/2.5 x 10(6) cell) cell-associated cytokine expression was induced by hemofiltration. DISCUSSION AND CONCLUSION: Polyamide hemofiltration is a highly biocompatible technique due to the use of a synthetic membrane with a sterile reinfusion fluid and the convective removal of the activated anaphylatoxins and Beta 2 microglobulin. PMID- 9745997 TI - Mechanical circulatory support after orthotopic heart transplantation. AB - Frequently the only therapy for primary graft- and right heart failure, as well as low output syndrome from acute of chronic rejection, is implantation of a mechanical circulatory support system, until recompensation or retransplantation. At our institution, mechanical assist devices were implanted in 25 heart recipients for a cute rejection (n=9), primary graft failure (n=7), acute right heart failure (n=7), and chronic rejection with low output syndrome (n=2). Patients (pts) with primary graft failure (n=3) received an intraaortic balloon pump (IABP), one pt an IABP plus Abiomed-System for left ventricular support, one pt the Thoratec-System for biventricular support. Patients with right heart failure (RHF) received the Biomedicus centrifugal pump for right ventricular support. Nine pts suffered from acute rejection. Six pts received an IABP, one the Biomedicus as femoro-femoral bypass, one the Abiomed-System for biventricular support, two the Thoratec-System for biventricular support and two within this group switched from the Biomedicus pump to the Thoratec-System for biventricular support. Patients with chronic graft failure (n=2) received the Novacor-System (LVAD) for left ventricular support, one received a Tojobo-System and an oxygenator for biventricular support post coronary artery bypass surgery. Support time ranged from 0.5-h to 73 days. Five pts were weaned. Two (8%) of 25 pts were retransplanted, 18 (72%) died in spite of mechanical support from multiple organ failure. The use of a mechanical assist device after heart transplantation is encouraging only in the case of early right heart failure, as well as primary and chronic graft failure. In view of the poor results, the use of mechanical assist devices should not be recommended in the case of heart failure caused by acute rejection. PMID- 9745998 TI - Development and initial in vivo testing of a new hydraulic drive system (Paedipump) for circulatory support in infants. AB - The main limitation in the use of circulatory support in children is the lack of an adequate system with regard to size and pumping capacity. Recently, two pneumatically driven ventricular support systems with low volume chambers for use in a pediatric population became available. We have developed a hydraulic drive system with an advantageous exact control of the stroke volume. The system enables two different modes of operation: the full-empty and the filled-empty modes. In both cases the ventricle is empty at the end of systole. This new system was tested in experimental animals (6 pigs, body weight 9.5-14.0 kg) with normal and reduced left ventricular function (MAP<45 mmHg). A 25 ml ventricle (HIA-Medos) was implanted. The full-empty and the filled-empty mode used led to a significant load reduction, both in animals with normal and impaired cardiac function. Plasma lactate levels, pH-values and total body O2-consumption were in the normal range during circulatory support indicating adequate organ perfusion. Results showed that sufficient ventricular support was achieved during all pumping modes due to the possibility of controlling and modifying the stroke volume of the hydraulically driven support system employed according to necessity. This is a promising feature for its future application in infants with congenital or acquired heart diseases. PMID- 9745999 TI - Long-distance transportation of patients with a paracorporeal left ventricular assist device. AB - Patients in cardiogenic shock refractory to basic medical therapy may require specialised assistance in a multispeciality environment distant to the institution where they are first admitted. In such situations, transportation of these critical patients may be difficult and involve many risks. Here we describe the case histories of two patients who underwent implantation of a paracorporeal left ventricular assist device and were transferred to specialised institutions for extended treatment. The distance of transportation was 400 km and the patients were transported by ambulance and helicopter. Some aspects of logistics and complexity of long-distance transportation are also commented on. PMID- 9746000 TI - Anemia and blood pressure correction obtained by daily hemodialysis induce a reduction of left ventricular hypertrophy in dialysed patients. PMID- 9746001 TI - A treatment that works. PMID- 9746002 TI - Epileptic aphasia: a consequence of regional hypometabolic encephalopathy? AB - A series of 25 children, 13 females and 12 males, who had an acquired communication disorder together with epilepsy, but did not fulfil the strict criteria of the Landau-Kleffner syndrome, was studied. All children had a clinical neurological evaluation, speech and language assessment, an awake and sleep EEG, cranial MRI, SPET scan, and audiometry. Clinical seizures were most often polymorphic in type (17 of 25). Atypical absences were the commonest individual seizure type occurring in 15 cases. All patients had an unequivocal epileptiform EEG. Normal sleep phenomena were only observed in 10 cases, enhancement of epileptiform activity in sleep was seen in 16. Cranial MRI was abnormal in six and normal in 19 cases. The SPET scans were abnormal in 22 of 25 children. The language deficits were classified neurologically as receptive aphasia, 24 of 25; expressive aphasia, 20 of 25; nominal aphasia, eight of 25; articulatory dyspraxia, 10 of 25; and auditory agnosia, nine of 25. It is hypothesized that the loss of communication skills is due to an encephalopathy secondary to the persistent epileptic discharge and manifests as a hypometabolic area on the SPET scan. PMID- 9746003 TI - Inappropriate use of carbamazepine and vigabatrin in typical absence seizures. AB - Carbamazepine and vigabatrin are contraindicated in typical absence seizures. Of 18 consecutive referrals of children with resistant typical absences only, eight were erroneously treated with carbamazepine either as monotherapy or as an add on. Vigabatrin was also used in the treatment of two children. Frequency of absences increased in four children treated with carbamazepine and two of these developed myoclonic jerks, which resolved on withdrawal of carbamazepine. Absences were aggravated in both cases where vigabatrin was added on to concurrent treatment. Optimal control of the absences was achieved with sodium valproate, lamotrigine, or ethosuximide alone or in combination. PMID- 9746004 TI - What constitutes cerebral palsy? AB - Cerebral palsy (CP) is a term of convenience applied to a group of motor disorders of central origin defined by clinical description. It is not a diagnosis in that its application infers nothing about pathology, aetiology, or prognosis. It is an umbrella term covering a wide range of cerebral disorders which result in childhood motor impairment. The precise inclusion criteria vary with the objectives for using the term. For meaningful comparison of rates of CP, as performed by and between CP registers, it is important that the rates should be generated using the same criteria. As generally understood there must be motor impairment, and this impairment must stem from a malfunction of the brain (rather than spinal cord or muscles). Furthermore, the brain malfunction must be non progressive and it must be manifest early in life. For the purposes of comparisons of rates across time even when the condition meets all the above criteria, it must not historically have been excluded from the category of CP. This paper addresses the problem of standardizing the inclusion criteria for selecting people included on CP registers with particular reference to this last criterion. PMID- 9746005 TI - Kinematic and kinetic analysis of running in children with cerebral palsy. AB - Computer-based analysis of gait was used to study walking and running in 19 children with spastic-diplegic cerebral palsy (CP) and 15 healthy control children. Temporospatial parameters, kinematic and kinetic data were compared and contrasted between groups for both types of gait. The majority of children with diplegic CP, who are independent ambulators, are able to run. These children increase their velocity by increasing their cadence, a mechanism that is distinct (and presumably less energy efficient) from that used by healthy children. Sagittal-plane kinematic and kinetic profiles at the ankle in children with CP were more similar to normal profiles in running than in walking, suggesting that the primary deviations at the ankle associated with CP are better tolerated at greater velocities. Relative power analysis showed that, like healthy children, those with CP depend more upon the proximal musculature about the hip for power generation as the velocity of gait increases. Children with CP achieve energy transfer between adjacent joints during walking and running in a manner comparable to unaffected children. Running is an important activity for children and should be considered in the functional assessment of those with CP. PMID- 9746006 TI - Cognitive deficits associated with frontal-lobe infarction in children with sickle cell disease. AB - This study examined the cognitive manifestations of frontal-lobe infarction in a population of children with sickle cell disease (SCD). Forty-one patients with SCD underwent MRI. Five patients with stroke symptoms had large infarcts encroaching on the tissue of the frontal lobes. Four patients without symptoms had smaller frontal-lobe infarcts. The patients with stroke were significantly impaired on measures of intelligence, memory, and frontal-lobe function (Wisconsin Card Sorting Test, WCST) compared with both the patients with normal MRI scans (N=30) and a group of sibling controls (N=15), who did not differ from each other. Patients with covert infarction obtained scores on the intelligence tests and the WCST that fell in between those of the stroke patients and the other two groups. This trend toward impairment suggests that patients with covert infarction are at similar risk for cognitive deficits to those with stroke. PMID- 9746007 TI - Childhood headaches: discrete entities or continuum? AB - The aims of this study were to investigate the signs and symptoms of recurrent headaches in children and to identify if there are any discrete groups of children whose headaches corresponded to the World Federation of Neurology (1969) definition of migraine. One-hundred and fifty children recruited from the neurology clinics at Royal Manchester, Booth Hall, and Birmingham Children's Hospitals were interviewed to complete a standardized questionnaire. The data were examined using cluster analysis followed by comparative analysis of the headache signs and symptoms in the different groups identified. No stable groups were identified (i.e., no group was reliably identified by different methods of cluster analysis) which corresponded to the World Federation of Neurology definition of migraine. This would suggest that this definition is not appropriate for the sample investigated. Three groups of children evolved after cluster analysis. None of these groups was in agreement with the International Headache Society classification of headaches. The groups were neither 'stable' clusters nor 'useful' in predicting prognosis. This outcome supports the continuum theory of headache syndromes. PMID- 9746008 TI - Effects of fluoxetine treatment in young children with idiopathic autism. AB - Thirty-seven children, aged between 2 and 7 years, with idiopathic autism underwent an open-label trial of fluoxetine treatment. All had assessment of diagnosis, developmental status, and family psychiatric history. Independent developmental testing before and after starting fluoxetine permitted quantification of language acquisition in a subgroup. Twenty-two of the 37 children had a beneficial treatment response sustained during continuing treatment for 13 to 33 months (mean 21 months). Eleven had an excellent response and were able to attend mainstream classrooms. Eleven had a good response though they remained identifiably autistic. Fifteen children had no benefit. Responders showed behavioral, language, cognitive, affective, and social improvements. Responders with adequate testing showed marked increases in language acquisition at every stage of development as compared with (1) pretreatment status, (2) responses to other treatments, (3) ability in non-language (matching) tasks, and (4) historical controls from the literature. The response to fluoxetine strongly correlated with a family history of major affective disorder. These preliminary findings implicate serotonergic mechanisms in autistic symptomatology and warrant further study with controlled trials. PMID- 9746009 TI - Development of visual function in hemihydranencephaly. AB - This study reports on the findings of longitudinal follow-up of visual function in a 12-year-old girl affected by congenital right hemihydranencephaly. This extremely rare unilateral brain malformation allowed the authors to gather new information on neuronal plasticity and functional compensations of the visual system across a period of 10 years. An extension of the preserved right visual hemifields above the middle line and strategical eye or head positions developed to increase visual functions are discussed. In addition, ophthalmological and orthoptical findings, as well as the development of monocular grating and linear acuity, are described. PMID- 9746010 TI - A case of Ohtahara syndrome with cytochrome oxidase deficiency. AB - Ohtahara syndrome is a rare cause of epileptic seizures during the neonatal period. This is believed to be the first report of this syndrome with a specific metabolic defect. Defects in respiratory chain function may be more common than previously assumed in patients with this epilepsy syndrome. PMID- 9746011 TI - White-matter disease of prematurity, periventricular leukomalacia, and ischemic lesions. PMID- 9746012 TI - Current articles. PMID- 9746013 TI - New car, no keys. PMID- 9746014 TI - Can quality of clinical trials and meta-analyses be quantified? PMID- 9746015 TI - Streptogramins: an answer to antibiotic resistance in gram-positive bacteria. PMID- 9746016 TI - Neuropathy complicating diffuse infiltrative lymphocytosis. PMID- 9746017 TI - Not much progress in treatment of cerebral malaria. PMID- 9746018 TI - Has the target autoantigen for Graves' ophthalmopathy been found? PMID- 9746019 TI - International multicentre pooled analysis of late postnatal mother-to-child transmission of HIV-1 infection. Ghent International Working Group on Mother-to Child Transmission of HIV. AB - BACKGROUND: An understanding of the risk and timing of mother-to-child transmission of HIV-1 in the postnatal period is important for the development of public-health strategies. We aimed to estimate the rate and timing of late postnatal transmission of HIV-1. METHODS: We did an international multicentre pooled analysis of individual data from prospective cohort studies of children followed-up from birth born to HIV-1-infected mothers. We enrolled all uninfected children confirmed by HIV-1-DNA PCR, HIV-1 serology, or both. Late postnatal transmission was taken to have occurred if a child later became infected. We calculated duration of follow-up for non-infected children from the time of negative diagnosis to the date of the last laboratory follow-up, or for infected children to the mid-point between the date of last negative and first positive results. We stratified the analysis for breastfeeding. FINDINGS: Less than 5% of the 2807 children in four studies from industrialised countries (USA, Switzerland, France, and Europe) were breastfed and no HIV-1 infection was diagnosed. By contrast, late postnatal transmission occurred in 49 (5%) of 902 children in four cohorts from developing countries, in which breastfeeding was the norm (Rwanda [Butare and Kigali], Ivory Coast, Kenya), with an overall estimated risk of 3.2 per 100 child-years of breastfeeding follow-up (95% CI 3.1 3.8), with similar estimates in individual studies (p=0.10). Exact information on timing of infection and duration of breastfeeding was available for 20 of the 49 children with late postnatal transmission. We took transmission to have occurred midway between last negative and first positive HIV-1 tests. If breastfeeding had stopped at age 4 months transmission would have occurred in no infants, and in three if it had stopped at 6 months. INTERPRETATION: Risk of late postnatal transmission is consistently shown to be substantial for breastfed children born to HIV-1-positive mothers. This risk should be balanced against the effect of early weaning on infant mortality and morbidity and maternal fertility. PMID- 9746020 TI - T axis as an indicator of risk of cardiac events in elderly people. AB - BACKGROUND: The T axis was postulated to be a general marker of repolarisation abnormality, indicative of subclinical myocardial damage. The aim of this investigation was to assess the prognostic importance of the T axis for fatal and non-fatal cardiac events, in a prospective cohort study of men and women aged 55 years and older. METHODS: 2352 men and 3429 women from the population-based Rotterdam Study took part in the study. Electrocardiograms were done, and T axes were categorised as normal, borderline, or abnormal. Data were analysed with Cox's proportional-hazards models; adjustment for age and sex was done where appropriate. FINDINGS: During 3-6 (mean 4) years of follow-up of the 5781 participants, 165 (2.9%) fatal and 192 (3.3%) non-fatal cardiac events occurred. Participants with an abnormal T axis (n=609) had an increased risk of cardiac death (hazard ratio 3.9 [95% CI 2.8-5.6]), sudden cardiac death (4.4 [2.6-7.4]), non-fatal cardiac events (2.7 [1.9-3.9]), and combined fatal or non-fatal cardiac events (3.2 [2.5-4.1]); p<0.001 for each. Additional adjustment for established cardiovascular risk factors resulted in lower, but still significant risk for all endpoints. The risk associated with an abnormal T axis was higher than those for any other cardiovascular risk factor. Additional subgroup analyses indicated that the risk of cardiac death was not substantially modified by age, sex, or history of myocardial infarction. INTERPRETATION: The T axis is a strong and independent risk indicator of fatal and non-fatal cardiac events in the elderly. PMID- 9746021 TI - Socioeconomic status, standard of living, and neurotic disorder. AB - BACKGROUND: Evidence on the association between socioeconomic status and the prevalence of neurotic disorder is contradictory. We studied the association between three elements of socioeconomic status and the prevalence of neurotic psychiatric disorder in a representative sample of adults aged 16-64 living in private households in the UK. METHODS: A cross-sectional survey of 10,108 adults aged 16-65 resident in private households in the UK was selected by a multi stage, clustered, random-sampling design. Neurotic disorders were defined using a standardised interview, the revised clinical interview schedule (CIS-R). Data for 9570 people were available for this study. FINDINGS: We used housing tenure and access to cars as measures of standard of living; both were associated with the prevalence of neurotic disorder even after adjustment for other socioeconomic and demographic variables, including Registrar General's Social Class and educational attainment. Those people with no access to a car had an odds ratio for neurotic disorder of 1.4 (95% CI 1.1-1.7), compared with those who had access to two or more cars. People who rented their homes were also at increased risk (1.3 [1.1 1.5]). We estimated that about 10% of the neurotic disorder in the UK could be attributed to the increased prevalence of those without cars who rented their homes. There was a complex interaction between Registrar General's Social Class and sex, and there was no independent association with educational attainment. INTERPRETATION: There is an independent association between low standard of living and the prevalence of neurotic psychiatric disorder. The UK has experienced one of the largest increases in income inequality within western market economies over the past 20 years, and this inequality may have had adverse consequences for the mental health of the population. PMID- 9746022 TI - Does quality of reports of randomised trials affect estimates of intervention efficacy reported in meta-analyses? AB - BACKGROUND: Few meta-analyses of randomised trials assess the quality of the studies included. Yet there is increasing evidence that trial quality can affect estimates of intervention efficacy. We investigated whether different methods of quality assessment provide different estimates of intervention efficacy evaluated in randomised controlled trials (RCTs). METHODS: We randomly selected 11 meta analyses that involved 127 RCTs on the efficacy of interventions used for circulatory and digestive diseases, mental health, and pregnancy and childbirth. We replicated all the meta-analyses using published data from the primary studies. The quality of reporting of all 127 clinical trials was assessed by means of component and scale approaches. To explore the effects of quality on the quantitative results, we examined the effects of different methods of incorporating quality scores (sensitivity analysis and quality weights) on the results of the meta-analyses. FINDINGS: The quality of trials was low. Masked assessments provided significantly higher scores than unmasked assessments (mean 2.74 [SD 1.10] vs 2.55 [1.20]). Low-quality trials (score < or = 2), compared with high-quality trials (score > 2), were associated with an increased estimate of benefit of 34% (ratio of odds ratios [ROR] 0.66 [95% CI 0.52-0.83]). Trials that used inadequate allocation concealment, compared with those that used adequate methods, were also associated with an increased estimate of benefit (37%; ROR=0.63 [0.45-0.88]). The average treatment benefit was 39% (odds ratio [OR] 0.61 [0.57-0.65]) for all trials, 52% (OR 0.48 [0.43-0.54]) for low-quality trials, and 29% (OR 0.71 [0.65-0.77]) for high-quality trials. Use of all the trial scores as quality weights reduced the effects to 35% (OR 0.65 [0.59-0.71]) and resulted in the least statistical heterogeneity. INTERPRETATION: Studies of low methodological quality in which the estimate of quality is incorporated into the meta-analyses can alter the interpretation of the benefit of intervention, whether a scale or component approach is used in the assessment of trial quality. PMID- 9746023 TI - Psychological effect of witnessed resuscitation on bereaved relatives. AB - BACKGROUND: Established practice is for the relatives of critically ill patients to be excluded from the clinical area during resuscitation. We aimed to discover whether relatives wanted to be present during the resuscitation of a family member and whether witnessing resuscitation had any adverse psychological effects on bereaved relatives. METHODS: In this pilot study, relatives of patients who required resuscitation were given the option to remain with the patient during resuscitation or were not given this choice and directed to the relatives' room (control group). The unit of randomisation was the patient who required resuscitation and not the relatives. One close relative was paired with each patient. All relatives were accompanied by a chaperone who gave emotional support and provided technical information on the resuscitation. Relatives were followed up 1 month after the resuscitation. We used a questionnaire to ask about the decision to be present or absent during resuscitation. Bereaved relatives also completed five standardised psychological questionnaires to assess anxiety, depression, grief, intrusive imagery, and avoidance behaviour. FINDINGS: 25 patients underwent resuscitation (13 in witnessed resuscitation group, 12 in control group). Three patients in the witnessed group survived, all the control group patients died. Two relatives in each group were lost to follow-up. Thus, eight relatives who witnessed resuscitation and ten control-group relatives were followed up. There were no reported adverse psychological effects among the relatives who witnessed resuscitation, all of whom were satisfied with their decision to remain with the patient. The clinical team became convinced of the benefits to relatives of allowing them to witness resuscitation if they wished, so the trial was terminated. INTERPRETATION: In the context of the emergency department, routine exclusion of relatives from the resuscitation room may no longer be appropriate. PMID- 9746024 TI - A young woman with petechiae. PMID- 9746025 TI - Fatal aplastic anaemia associated with nifedipine. PMID- 9746026 TI - Intranasal midazolam for childhood seizures. PMID- 9746027 TI - Gamma-band electroencephalographic oscillations in a patient with somatic hallucinations. PMID- 9746028 TI - p53 mutations in BRCA1-associated familial breast cancer. PMID- 9746029 TI - Persistence of Borna disease virus-specific nucleic acid in blood of psychiatric patient. PMID- 9746030 TI - Obstinate imitation behaviour in differentiation of frontotemporal dementia from Alzheimer's disease. PMID- 9746031 TI - Long-term survival of homocystinuria: the first case. PMID- 9746032 TI - Relief by botulinum toxin of voiding dysfunction due to prostatitis. PMID- 9746033 TI - Enterohaemorrhagic Escherichia coli in Ngoila (Cameroon) during an outbreak of bloody diarrhoea. PMID- 9746034 TI - Breast cancer in female flight attendants. PMID- 9746035 TI - Value of HRT in women with heart disease doubted. PMID- 9746036 TI - Children are not small adults. PMID- 9746037 TI - Japanese victims of discrimination against lepers sue government. PMID- 9746038 TI - Women's health issues cause controversy in European Union. PMID- 9746039 TI - Terrorist attack in Northern Ireland prompts major emergency action. PMID- 9746040 TI - Guillain-Barre syndrome. AB - Guillain-Barre syndrome (GBS) is viewed as a reactive, self-limited, autoimmune disease triggered by a preceding bacterial or viral infection. Campylobacter jejuni, a major cause of bacterial gastroenteritis worldwide, is the most frequent antecedent pathogen. It is likely that immune responses directed towards the infecting organisms are involved in the pathogenesis of GBS by cross-reaction with neural tissues. The infecting organism induces humoral and cellular immune responses that, because of the sharing of homologous epitopes (molecular mimicry), cross-react with ganglioside surface components of peripheral nerves. Immune reactions against target epitopes in Schwann-cell surface membrane or myelin result in acute inflammatory demyelinating neuropathy (85% of cases); reactions against epitopes contained in the axonal membrane cause the acute axonal forms of GBS (15% of cases). Care for such patients may be challenging, yet the prognosis overall is favourable. Optimal supportive care and anticipation and prevention of complications are the mainstay of therapy. Admission to the intensive-care unit is necessary in 33% of patients who require intubation and assisted ventilation. Immunomodulation with infusions of IgG or plasma exchange treatments foreshorten the disease course. PMID- 9746041 TI - The healer and the healed: works and life of Kenzaburo Oe. PMID- 9746042 TI - Peptic ulcer: Moynihan's or Marshall's disease? PMID- 9746043 TI - Registering refugee and asylum-seeking doctors. PMID- 9746044 TI - Chronic obstructive pulmonary disease: don't forget the gatekeeper. PMID- 9746045 TI - Sexually transmitted diseases. PMID- 9746046 TI - Sexually transmitted diseases. PMID- 9746048 TI - Sexually transmitted diseases. PMID- 9746047 TI - Sexually transmitted diseases. PMID- 9746049 TI - Opiates for sickle-cell crisis. PMID- 9746050 TI - Airline travel in sickle-cell disease. PMID- 9746051 TI - Magnetic-resonance imaging and breast cancer multicentricity. PMID- 9746053 TI - Vitamin A supplementation and HIV-1 mother-to-child transmission in Africa. PMID- 9746052 TI - Vitamin A supplementation and HIV-1 mother-to-child transmission in Africa. PMID- 9746054 TI - Wernicke's encephalopathy. PMID- 9746055 TI - Evidence on vitamin B6 questioned. PMID- 9746056 TI - Evidence on vitamin B6 questioned. PMID- 9746057 TI - Morbidity from renovascular disease in elderly patients with congestive cardiac failure. PMID- 9746058 TI - Recommendations after the RAGE litigation. Radiation Action Group Exposure. PMID- 9746059 TI - Local surveillance of antimicrobial resistance. Synercid Resistance Surveillance Group. PMID- 9746060 TI - Postpartum coma. PMID- 9746061 TI - Audit and use of NSAIDs. PMID- 9746064 TI - Identification of antigenic sites on three hepatitis C virus proteins using phage displayed peptide libraries. AB - A novel approach to screening phage-displayed peptide libraries has been used to identify hepatitis C virus (HCV) core, NS4 and NS5 sequences, which are antigenic in humans. Two random peptide libraries were used for screening using a mixture of HCV-positive sera or individual antibodies to core, NS3, NS4, and NS5 HCV proteins affinity-purified from this mixture. Sequencing of 56 selected phage clones resulted in 28 different peptide sequences and identification of seven antigenic regions, three in the core protein (19-26, 34-49, and 73-83), three in the NS4 (1681-1693, 1712-1718, and 1726-1736) and one in the NS5 protein (2251 2260). No NS3-specific peptides were identified. The immune response to core, NS4 and NS5 proteins includes a variety of linear determinants whereas epitopes on the investigated part of NS3 protein appear to be conformation-dependent. PMID- 9746065 TI - Interleukin 12 enhances deficient HCV-antigen-induced Th1-type immune response of peripheral blood mononuclear cells. AB - The aim of this study was to examine the possible immunomodulating effects of rhIL-12 on the immune response induced by different hepatitis C virus (HCV) antigens. Freshly isolated peripheral blood mononuclear cells (PBMC) of 33 patients with chronic HCV infection were stimulated with optimal concentrations of antigens from the NS3, NS4, NS5, and core region of HCV in the absence or presence of interleukin-12 (IL-12). Stimulation by alpha-CD3 + alpha-CD28, lipopolysaccharide (LPS), and pokeweed mitogen (PWM) were used as controls. Proliferation and cytokine production were determined by 3H-thymidine uptake and enzyme-linked immunosorbent assay (ELISA) after 72 hr. After stimulation with antigen or antigen + IL-12, increased proliferation and production of interferon gamma (IFN gamma) and tumor necrosis factor-alpha (TNF alpha) were observed in 23 of the 33 patients. Thus, a separation of the patients into HCV-antigen/IL-12 responders (group 1, n = 23) and HCV-antigen/IL-12 nonresponders (group 2, n = 10) was possible. Lower baseline IL-12- and LPS-induced IFN gamma, TNF alpha, and IL-12 production was observed in group 2 due to a possible dysfunction of accessory cells. Significant antigen-induced Th2-type cytokine (IL-4, IL-10, IL 13) production was not found. According to clinical and serological parameters, group 2 comprised mostly patients with advanced liver disease. These data suggest an HCV-related cellular immune defect in patients with hepatitis C that can be restored in most patients by IL-12. PMID- 9746066 TI - Prevalence, implication, and viral nucleotide sequence analysis of GB virus C/hepatitis G virus infection in acute fulminant and nonfulminant hepatitis. AB - The clinical impact of GB virus-C (GBV-C)/hepatitis G virus (HGV) infection on various causes of acute hepatitis and fulminant hepatitis is controversial. In this study, serum samples from 164 patients with acute hepatitis of various causes, 34 asymptomatic hepatitis B virus (HBV) carriers, and 34 healthy adults were tested for GBV-C/HGV RNA by reverse transcription-nested polymerase chain reaction using primers based on the 5'-untranslated region. Nucleotide sequences of GBV-C/HGV RNA from various groups were compared. The prevalence of GBV-C/HGV RNA was significantly higher in patients with acute hepatitis D virus (HDV) superinfection than in HBV carriers or healthy controls (10/37 vs. 2/34, P < 0.02; 10/37 vs. 1/34, P < 0.005). GBV-C/HGV RNA was detected in 11.1% of acute hepatitis A patients, 9.5% of acute hepatitis B patients, 15.8% of acute hepatitis C patients, 12.5% of acute hepatitis E patients, 11.8% of chronic hepatitis B patients with acute exacerbation, and 11.1% in patients with non-A to -E hepatitis; each was not significantly higher than that in HBV carriers or healthy adults. There were no significant differences in gender, age, serum albumin, bilirubin, and alanine aminotransferase levels nor in the occurrence of fulminant hepatitis (6/28 vs. 36/136) between patients with or without GBV-C/HGV RNA. All six patients with fulminant hepatitis who had GBV-C/HGV RNA were complicated by infection with hepatitis B, C, or D. The GBV-C/HGV clones from 21 patients with or without fulminant hepatitis belonged to group 3. No particular strain of GBV-C/HGV was associated with fulminant hepatitis. PMID- 9746067 TI - High prevalence of GB virus C/hepatitis G virus infection among homosexual men infected with human immunodeficiency virus type 1: evidence for sexual transmission. AB - GB virus C/hepatitis G virus (GBV-C/HGV), a recently discovered orphan flavivirus, is distantly related to hepatitis C virus (HCV). Although both GBV C/HGV and HCV can be transmitted by the parenteral route, their principal modes of transmission and associated risk behaviors may differ. Using reverse transcription-polymerase chain reaction, the 5'-noncoding regions of GBV-C/HGV and HCV were amplified from plasma or sera of 209 individuals infected with human immunodeficiency virus type 1 (HIV-1). As verified by Southern blot analysis, GBV C/HGV and HCV infection were detected in 37 (17.7%) and 22 (10.5%) of 209 HIV-1 infected individuals, respectively. GBV-C/HGV infection was significantly associated with homosexual sex (P = 0.044) and was more common than HCV infection among HIV-1-infected homosexual men (P = 0.006). The prevalence of GBV-C/HGV infection was nearly equal in women infected with HIV-1 via high-risk heterosexual sex (14.0%) or injection drug use (IDU) (17.5%). By contrast, HCV infection was associated significantly with women reporting IDU when compared to women reporting high-risk heterosexual sex (P < 0.0001). Alanine aminotransferase levels were elevated in HIV-1-infected individuals who were co-infected with HCV (P = 0.009), but not with GBV-C/HGV (P = 0.9). The high prevalence of GBV-C/HGV infection in HIV-1-infected nondrug-injecting homosexual men and among women engaging in high-risk heterosexual sex is consistent with transmission by the mucosal route and with acquisition of infection by the receptive rather than insertive partner. PMID- 9746068 TI - Fecal excretion of a nonenveloped DNA virus (TTV) associated with posttransfusion non-A-G hepatitis. AB - Five patients with type B or C hepatocellular carcinoma were found to be infected with a nonenveloped DNA virus (TTV) associated with posttransfusion hepatitis of non-A-G etiology. Paired feces and serum samples from these patients were tested for TTV DNA by polymerase chain reaction with seminested primers and their sequences were compared. TTV DNA was detected in sera from all of the patients, while it was detected in feces from three patients, including two with high viral titers in serum. When feces and serum from one patient were subjected to floatation ultracentrifugation in CsCl, TTV in feces banded at a peak density of 1.35 g/cm3 and that in serum at 1.31-1.32 g/cm3. TTV isolates in three pairs of feces and serum had the identical sequence of 222 base pairs. The excretion of TTV into feces indicates that TTV would be transmitted not only parenterally but also nonparenterally by a fecal-oral route. PMID- 9746069 TI - Evaluation of the detection of human papillomavirus genotypes in cervical specimens by hybrid capture as screening for precancerous lesions in HIV-positive women. AB - Given the frequency and persistence of human papillomavirus (HPV) infection and associated cytological alterations in HIV-1-positive women, the incidence of uterine cervix neoplasm is likely to increase along with patient survival. More appropriate screening programs, which, in addition to Pap smears (PS), also include tests to detect and type HPV, are needed for the early identification of precancerous cervical lesions. This prospective study involved 168 HIV-positive (group A) and 100 HIV-negative women (group B). Cervicovaginal samples were collected for a PS and HPV DNA search. The detected virus was typed as high intermediate oncogenic risk HPV (HR-HPV) and low-risk HPV (LR-HPV) using hybrid capture (HC) (Murex-Digene) and in-house PCR tests. The HC-detected prevalence of HPV was 111/168 (66%:HR 75.6%) in group A and 15/100 (15%:HR 42.9%) in group B (P < 0.0001). Polymerase chain reaction (PCR) was positive in 91% and 48%, respectively. No significant difference was observed between drug addicts and heterosexual HIV-1-positive women (P = 0.09). HPV was detected in 94% of the 57 HIV-positive women with cytological alterations. HR-HPV was found in 41/49 women with low-grade and 7/8 with high-grade squamous intraepithelial lesions (LSIL and HSIL, respectively). In women with a negative PS, HPV was detected in 57/111 cases (HR 63%) of group A and in 13/98 of group B (6 cases of HR). Of the 54 group A women who underwent biopsy, histology revealed that 41 had LSIL (18 with negative PS, 19 with LSIL, and 4 with HSIL; HR-HPV in 73% and LR-HPV in 17%), nine had HSIL (5 LSIL and 4 HSIL on cytology; HR-HPV in 89% and LR-HPV in 11%), and four were negative (all cytology negative; 3 HR-HPV and 1 LR-HPV). HR-HPV was more frequent as immunodepression worsened. These results show that cytological evaluation alone underestimated histological alterations in 23/50 women (42.6%), whereas the combination of Pap smear and HPV detection reduced this underestimate to 5%. PMID- 9746070 TI - False negativity by an anti-HIV assay kit (IMx 8B32) and evaluation of its replacement (IMx 8C98). AB - False negativity in a commercial anti-HIV kit (IMx HIV-1/HIV-2 3rd Generation Plus (code 8B32) was investigated, and the kit that superseded it (IMx HIV-1/HIV 2 III Plus, code 8C98) was evaluated. In a comparison on 574 freshly collected anti-HIV-1-positive specimens, 97.2% were more reactive in 8C98 than in 8B32; 35.5% were more than twice as reactive and 8.5% were more than four times as reactive. In 8B32, the signal from 55 specimens selected because of weak reactivity was enhanced 1.5 to 8.8 times by preliminary heating at 56 degrees C for 30 min. The reactivity of the 55 heated sera was then similar to that of the same specimens tested without heat treatment in the 8C98 assay. Reactivity in 8B32 was also increased in 66 of 76 (at least twofold in 20) randomly chosen anti HIV-positive serum specimens by the addition of EDTA (10 mM final concentration). One of these specimens was false negative (signal:cutoff (S:CO) ratio 0.76) in 8B32, though its reactivity was restored by addition of EDTA (S:CO ratio 9.54). These findings indicate that the inhibitory effect that originally led to false negative findings in 8B32 was probably due to complement activity, and that the same activity was present in the freshly collected specimens used here to evaluate the replacement IMx anti-HIV assay (8C98). The specimen panel employed to evaluate 8C98 included 1,892 anti-HIV-positive and 779 anti-HIV-negative specimens. There were no false negative reactions. The lowest S:CO ratio observed was 6.2 and only 17 (0.2%) anti-HIV-positive specimens gave ratios less than 10. Nine unreproducible false positive reactions arose, all possibly attributable to specimen carryover by the IMx instrument. The performance of 8C98 was also compared with that of 10 other current anti-HIV kits using 21 sets of seroconversion specimens (127 specimens in total), and five performance assessment panels (92 specimens in total) comprised mostly of single bleeds from recent seroconverters. IMx 8C98 was the second most sensitive assay. We found no evidence that the 8C98 kit was prone to the effect that had given rise to false negative results in its predecessor (8B32). PMID- 9746071 TI - Changes in intracellular cytokine levels in newborn and adult lymphocytes induced by HSV-1. AB - Changes in the expression of intracellular interleukin-2 (IL-2), interleukin-4 (IL-4), interferon (IFN)-gamma, and tumor necrosis factor (TNF)-alpha in newborn and adult lymphocytes induced by herpes simplex virus (HSV)-1 were examined. Cord blood mononuclear cells (CBMC) or adult peripheral blood mononuclear cells (PBMC) were infected with HSV-1 and cultured with phorbol 2-myristate 13-acetate (PMA) plus ionomycin in the presence of monensin for 4 hr. Surface antigen and intracellular cytokines were stained simultaneously and analyzed by flow cytometry. The percentage of cells that expressed IL-2, IFN-gamma, and TNF-alpha was significantly increased in HSV-1-infected CD3+, CD4+, CD8+, CD45RA+, and CD45RO+ lymphocytes compared with uninfected lymphocytes from adult PBMC. The percentage of cells that expressed IL-2 and TNF-alpha was increased significantly in HSV-1-infected CD3+, CD4+, CD8+, and CD45RA+ lymphocytes compared with uninfected lymphocytes from CBMC. IFN-gamma was under the detectable level in HSV 1-infected and uninfected lymphocytes from CBMC. Intracellular IL-4 was not detected in HSV-1 or in uninfected lymphocytes from PBMC and CBMC. These results demonstrate that HSV-1 enhances intracellular levels of IL-2, IFN-gamma, and TNF alpha in adult lymphocytes and defective IFN-gamma production in cord blood. PMID- 9746072 TI - Polymorphisms of thymidine kinase gene in herpes simplex virus type 1: analysis of clinical isolates from herpetic keratitis patients and laboratory strains. AB - Drug-resistance of herpes simplex virus (HSV) is caused most frequently by mutation of the viral thymidine kinase (TK) gene. To elucidate the significance of detecting nucleotide changes of the TK gene for screening drug-resistant viruses, the frequency and variation of the genetic polymorphisms in the whole coding region of the TK gene were studied in 14 acyclovir-susceptible HSV type 1 (HSV-1) clinical isolates from 14 patients with epithelial herpetic keratitis. Two reference HSV-1 laboratory strains, McKrae and PH, and two acyclovir resistant variants of the PH strain were also studied as controls. Polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) and direct sequencing detected nucleotide differences at 24 positions, and amino acid substitutions at 12 codons in the TK gene of the examined viruses. Nucleotide diversity of 0.0029 per base (the average number of nucleotide substitutions of 3.3 per 1,131 base pairs) in the TK gene in the clinical isolates was comparable to 0.0037 per base of the whole HSV-1 genome in Japanese isolates reported previously. PCR-SSCP analysis of the acyclovir-resistant strains easily detected aberrantly shifted bands by comparing them with those of the parental strain, followed by the quick determination of mutated sequences. These results suggest that detection of nucleotide changes of the TK gene is useful for serial observation of persistent or recurrent HSV infection as observed in immunocompromised hosts, but that it is not useful for screening drug-resistant viruses from nonepidemic clinical isolates because of the comparable genetic polymorphisms in the TK gene as in the whole HSV-1 genome. PMID- 9746073 TI - Development of viremia and humoral and cellular parameters of immune activation after vaccination with yellow fever virus strain 17D: a model of human flavivirus infection. AB - To monitor early and late events of immune system activation after primary and secondary flavivirus infection, 17 healthy persons were vaccinated with the standard 17D vaccine virus strain of yellow fever (YF). Twelve of these persons had not received YF vaccine previously and 5 had been vaccinated once at least 10 years before. Viremia and various parameters of humoral and cellular immune activation were followed daily for 7 days and weekly thereafter. Viremia was detected by reverse transcriptase-polymerase chain reaction in all 12 first-time vaccinees beginning from the second to the sixth day after vaccination; most tested positive between the fourth and sixth day. Infectious 17D virus was detected using a plaque forming assay in the serum of 7 of the 12 first-time vaccinees. As first parameters of immune activation, neopterin and beta2 microglobulin markedly increased between day 2 and day 6 postvaccination. In parallel to the viremia, circulating CD8+ T-cells significantly increased, with peak levels at day 5 after primary vaccination, indicating an activation of the cellular immune system. Neither viremia nor significant changes of these activation markers were observed in the five revaccinated persons. Neutralizing antibodies directed against the 17D vaccine strain developed in all persons within 2 weeks after vaccination. No correlation was found between the extent of viremia and the titer of neutralizing antibodies. Revaccination was followed by a minor and transient increase of neutralizing antibodies. High titers of neutralizing antibodies persisted for at least 10 years after primary vaccination. PMID- 9746074 TI - Comparison of RT-PCR with other diagnostic assays for rapid detection of influenza viruses. AB - To compare the effectiveness of reverse transcription-polymerase chain reaction (RT-PCR), shell vial culture and cytospin assay as laboratory techniques for rapid diagnosis of influenza infections, a retrospective study was carried out on 270 aliquots of oropharyngeal swabs collected from October 1993 to March 1996 and already characterized by standard isolation procedures, and a prospective study in which 65 clinical samples taken from patients with influenza-like syndrome between October 1996 and March 1997 were tested. In the retrospective study, using conventional isolation as the gold standard, the sensitivity of RT-PCR and cytospin assay for virus A was 100% (95% confidence interval (CI), 89.1-100) and for virus B it was 100% (95% CI, 56.1-100) compared with 77.5% (95% CI, 61.1 88.6) and 71.4% (95% CI, 30.3-94.9) for shell vial culture. The specificity of all the three assays was 100% (95% CI, 98.0-100) for virus A and 100% (95% CI, 98.2-100) for virus B. In the prospective study the sensitivity of RT-PCR was greater than that of the other tests considered, both rapid and standard. It is suggested that RT-PCR should be employed in combination with conventional culture techniques in routine diagnosis of influenza infections in order to obtain results more rapidly and to improve virus detection even in circumstances in which standard isolation could be problematic. PMID- 9746075 TI - Partial amplification of the measles virus nucleocapsid gene from stored sera and cerebrospinal fluids for molecular epidemiological studies. AB - The analysis of stored sera for retrospective molecular epidemiological studies provides a powerful tool to investigate strain variation in measles viruses that had circulated up to 20 years ago. For this purpose, a rapid and simple method for extraction of RNA from stored sera and cerebrospinal fluids (CSF) was developed. When used on sera and CSFs that have been frozen for as long as 20 years, this method proved to be more efficient than established techniques. The extracted RNA was reverse transcribed into cDNA by using random hexamer primers. The PCR amplification of the 3' terminus of the nucleocapsid gene (N) was divided into two overlapping fragments of 375 and 384 bp length, covering the entire region of interest. This region is thought to have the highest variability within the MV genome and has previously been shown to be suitable for strain characterization. The resulting PCR fragments were sequenced manually by using standard methods without the need of further clean-up steps. PMID- 9746076 TI - Mammography in the evaluation of masses in breasts reconstructed with TRAM flaps. AB - Patients with transverse rectus abdominis musculocutaneous (TRAM) flaps may develop physical findings, such as palpable masses, irregularities, and areas of increased tenderness that are suggestive of fat necrosis or recurrent malignancy. The purpose of this study was to evaluate these findings with mammography in an attempt to rule out recurrent malignancy in the autogenously reconstructed breast. Fifteen patients on whom mammography was performed as an aid in the evaluation of suspicious post-TRAM flap findings were reviewed. Common mammographic findings included calcifications believed to demonstrate fat necrosis, benign dermal calcifications, calcified hematoma, and clustered microcalcifications. Areas of increased or decreased density without calcifications were also identified and appeared to be related to surgical changes and fat necrosis. Twenty percent of patients had clustered microcalcifications, and 20% had detectable masses on mammography. One patient had a suspicious mass associated with clustered microcalcifications, leading to a biopsy that revealed fat necrosis. The majority of findings were consistent with normal fat within the TRAM flaps. This study supports mammography as a useful diagnostic tool in patients who have undergone TRAM flap breast reconstruction and who present postoperatively with suspicious physical findings. PMID- 9746077 TI - Reduction mammaplasty: its role in breast conservation surgery for early-stage breast cancer. AB - Segmental resection and radiotherapy is an accepted alternative over mastectomy for small, staged breast malignancies. However, women with large, pendulous breasts have been documented to have poorer cosmetic outcomes when undergoing irradiation after breast conservative surgery compared with women with small- or medium-size breasts, thought to be caused by dose inhomogeneity. The purpose of this study was to evaluate the efficacy of combining reduction mammaplasty with breast conservative surgery to facilitate postoperative irradiation. Between 1988 and 1996, 10 women have undergone bilateral reduction mammaplasty for breast malignancy followed by radiation therapy at our center. All women wished to avoid mastectomy (average age, 59 years). All lesions were detected preoperatively on mammography. The average amount of tissue removed was 945 g per breast. A variety of reduction techniques were employed to include the malignant lesions. All patients received 50 Gy of radiation therapy delivered in 25 fractions following reduction mammaplasty during a 5-week period. Radiation therapy was usually initiated within 4 weeks following surgery. Follow-up is currently 37 months, with all patients being followed for at least 8 months. No patients have had complications from the surgery or radiation therapy. No local recurrent malignancies have been detected. Cosmesis has been good to excellent in all patients. Despite equivalent survival outcomes for mastectomy for early-stage breast cancer, certain women are not good candidates for breast conservation and radiation therapy. An alternative for women with large, pendulous breasts that combines breast conservation therapy and concurrent bilateral reduction mammaplasty should be considered. This combination, in selected women, provides good functional and cosmetic results, and at the same time minimizes the potential difficulties of radiation therapy. PMID- 9746078 TI - Breast reduction improves symptoms of macromastia and has a long-lasting effect. AB - A retrospective study was undertaken to evaluate the long-term results of reduction mammaplasties. Of special interest was the extent to which the amount of breast tissue removed correlated with pain relief after reduction mammaplasty. One hundred fourteen patients underwent an average reduction of 1,266 g. The follow-up time was 7.7 years. Ninety-one percent of the patients noticed a decrease in shoulder, neck, and back pain, and lessening of brassiere shoulder grooves. Nine percent noticed no change at all, and only 1 patient complained of increased breast pain after the operation. In 11% of the patients, pain was not an indicator for the operation. We could find a significant correlation (r = 0.36, p = 0.001) between the amount of tissue resected and pain relief after surgery. The follow-up time did not correlate with a regaining of the physical complaints (r = 0.15, p = 0.2). Patients gained an average of 1.2 kg and had an average preoperative overweight of 118%. From the results of our study we were able to conclude that an average reduction of 500 to 600 g of breast tissue on both sides has a long-lasting effect in reducing shoulder and back pain. PMID- 9746079 TI - A simple mathematical formula for custom-made croissant tissue expanders. AB - The popularity of croissant-type tissue expanders has increased steadily during the past 5 years. Croissant-shaped expansion offers the advantage of creating a tailored skin flap that is advanced easily into an elliptical skin defect without the formation of dog-ears or without the need for backcuts in the expanded flap. Because the majority of lesions can be considered elliptical in shape, surgical removal is performed easily with an adapted croissant expander. We present a simple mathematical formula for calculating the exact dimensions of the required croissant expander. A good clinical result was obtained using this formula to design large custom-made croissant tissue expanders in a patient with a giant nevus. PMID- 9746080 TI - The antecubital fasciocutaneous island flap for elbow coverage. AB - Soft-tissue defects in the area of the periolecranon may be a source of concern to the reconstructive surgeon who aims for durable protection with a minimum of drawbacks. Lamberty and Cormack described the antecubital fasciocutaneous flap both as a local transposition and as a free flap. The island version of this flap enables a single-stage transfer of thin, pliable, sensitive skin into the region of the periolecranon without further scarring around the defect. In general, most of the donor site can be closed primarily together with a small, full-sheet, split-thickness skin graft on the remaining skin defect on the volar surface of the distal forearm. An additional advantage of this flap is the rather straightforward dissection with minimal repercussion on the forearm contour. An anatomic overview as well as 4 patients are described to illustrate the appealing features of this fasciocutaneous flap. PMID- 9746081 TI - Complications of mandibular fractures. AB - Timely repair of mandibular fractures remains an effective means to reduce pain, restore function, and prevent complications. This study addresses the effect of the time interval between injury and treatment on the overall complication rate, the complication rate between various treatment modalities (mandibular-maxillary fixation [MMF] alone, MMF with intraosseous wire bone fixation, and MMF with rigid internal fixation), and the relationship of inpatient vs. outpatient management. Cost was also examined with respect to choice of management. We report a retrospective series of 308 consecutive patients managed at the University of Miami/Jackson Memorial Hospital. Patients who received treatment 3 to 10 days following injury were found to have a lower complication rate than earlier or later repair. We postulate that most patients with mandibular fractures may be managed on an outpatient basis, which represents a considerable savings in cost. PMID- 9746082 TI - Prefabrication of a high-density porous polyethylene implant using a vascular induction technique. AB - Three-dimensional defects have been reconstructed with carved and remodeled frameworks wrapped within vascular carriers and have wide use in ear and nose reconstruction. The main problem with thick coverings is masking of the fine details in the frameworks. Other problems are insufficient blood supply, infection, and exposure of the implant. If sufficient vascular penetration can be established without any change in size and shape of the implant, it will be possible to cover it with a thin skin graft and improve aesthetic results. In this study we planned to prefabricate a high-density porous polyethylene implant that has been used frequently. The implants were placed and anchored underneath the superficial inferior epigastric artery and vein pedicle bilaterally in 10 adult New Zealand White rabbits. Twenty implants were prefabricated in this procedure, and they were evaluated via histological examination and perfusion scintigraphy. Results revealed that the implants were invaded by fibroneovascular tissue. Blood supply coming from the vascular pedicle was sufficient to maintain the implant as a prefabricated composite flap, which could be transferred as a pedicled flap or a free flap. PMID- 9746083 TI - Studies of surface microarchitecture using a hand-held video microscope in cases of cultured epithelial autografts. AB - The purpose of this study was to evaluate skin texture after cultured epithelial autografting (CEA) by focusing on the surface microarchitecture. From August 1993 through October 1995, CEA was performed to improve the texture of burn scars in 26 patients. The surface microarchitecture was studied using a hand-held video microscope on a long-term basis. In addition, biopsies were obtained from graft sites in 5 patients before CEA and at 1, 3, and 6 months after CEA. Before CEA, observation of the surface microarchitecture of the burn scars showed no skin fields or skin furrows. Histologically, the epidermis had no rete ridges. At 6 months after resurfacing with autologous cultured epithelium, it was possible to recognize skin fields and skin furrows in all patients. The shapes of the newly formed skin fields were similar but not identical to those at the donor site. Histological examination revealed that rete ridges were formed during the same time period. These findings suggest that the improvement in skin texture of burn scars after resurfacing with cultured epithelium depends on the reconstruction of the skin fields. PMID- 9746084 TI - Microcirculatory hemodynamics during the acute phase of free vascularized muscle allograft rejection. AB - This study investigated the microcirculatory pattern during acute rejection of a vascularized free muscle graft. Using a new rat cremaster muscle free flap model, isograft transplantation was done between genetically identical Lewis rats (N = 23) and allograft transplantation across a major histocompatibility barrier (Lewis Brown Norway [RT-11 + n] to Lewis [RT-11] rats; N = 24). At days 1, 3, 5, and 7 posttransplantation, microcirculatory measurements were taken of vessel diameter, red blood cell velocity, endothelial edema index, leukocytic endothelial interaction (rolling, adhering, and transmigrating leukocytes), and capillary perfusion. In the allografts at 5 days the lumen obstruction was 27% greater (p < 0.05) than onset. The number of perfused capillaries decreased by 36% compared with isografts at day 5 (p < 0.05). The number of adhering leukocytes increased significantly in allografts both at day 1 (71%; p < 0.05) and at day 3 (77%; p < 0.05). At day 3, transmigrating leukocytes rose significantly (p < 0.05). Allograft histology showed myocytic destruction and lymphocytic infiltration at day 5. Our results suggest that the microcirculatory signs of acute muscle allograft rejection occurred 24 hours after transplantation, and preceded histological signs of rejection. The critical time for microvascular survival of muscle allografts was 3 days. This procedure allows us to distinguish events related to transplantation trauma from rejection phenomena. PMID- 9746085 TI - Reconstruction of pharyngostomes with a modified deltopectoral flap combining endoscopy and tissue expansion. AB - The problem of pharyngostomic closure is difficult to solve, as evidenced by the large number of techniques described. The authors present the reconstruction of pharyngostomes by using Bakamjian's deltopectoral flap modified by the use of endoscopically introduced expanders for those patients in whom other techniques of choice (such as vascularized free flaps) have failed or are inapplicable either because of previous radiotherapy or because of local conditions. Bakamjian's deltopectoral flap, previously expanded with an expander coated with a partial-thickness skin graft and introduced endoscopically, allowed the authors to lift the flap in one operation to close the pharyngostome. This method provides the two walls of the pharynx (the skin graft as the inner aspect and the skin flap as the outer aspect), and the donor deltopectoral area is covered and epithelialized due to the skin graft. Thus by means of endoscopic expansion we use a nonaggressive technique to increase the surface area of the donor site and to increase its vascularization (delay phenomenon). Because the expander was coated with the graft, the authors were able to cover the anterior wall of the pharyngostome and the donor site in one surgical step. PMID- 9746086 TI - Laryngeal preservation surgery using a free flap patch following resection of a carcinoma of the posterior wall of the oropharynx. AB - A carcinoma originating from the posterior wall of the oropharynx is not common, and radiotherapy has been used for years in this event without acceptable success. A free flap patch was used in 4 patients to reconstruct the defect after resection of a T2 or T3 carcinoma on the posterior wall of the oropharynx without laryngectomy. The free flaps used were the radial forearm and the free jejunal patch in 2 patients each. There was no flap loss, and successful laryngeal preservation was obtained in 3 of 4 patients. Laryngeal preservation surgery using a free flap patch proved very useful in selected patients with carcinoma of the posterior wall of the oropharynx. Based on our clinical experience, the free jejunal patch seems superior to the free forearm flap with regard to postoperative functional results. PMID- 9746087 TI - Combination latissimus dorsi and rectus abdominis musculocutaneous flap for primary repair of a large upper limb defect. AB - The usefulness of flap transfer in reconstructive surgery is well established, and various types of flaps have been reported. However, concomitant use of skin grafts or repair by plural flaps must be utilized for large defects. We have obtained satisfactory results in the primary repair of a large defect covering the entire length of the upper limb by using a combination flap consisting of latissimus dorsi and rectus abdominis musculocutaneous flaps. This surgical procedure is relatively easy, safe, and reliable. Moreover, it is possible to harvest a combination flap possessing a wide arc of rotation with either the latissimus dorsi muscle or the rectus abdominis muscle as a pedicle. This reconstructive method should be considered for large defects. PMID- 9746088 TI - The sliding door flap for repair of vermilion defects. AB - The sliding door flap raised on the surface of the vermilion to the oral mucosa has been used for reconstruction of soft-tissue defects of the lower lip. The blood supply to this flap comes from the bilateral inferior labial artery. Utilizing both sides of the arteries, bilateral flaps allow for safe and easy transfer of the vermilion tissue to partial defects of the red lip. These flaps have been used in cases of lower lip defects with complete survival. The sliding door flap has increased mobility by at least 1 cm more than by the other style of flap reported by Goldstein. Moreover, this flap contributes to excellent cosmetic results. Our technique and clinical experiences are presented. PMID- 9746089 TI - The distally based ulnar artery forearm flap supplied by the dorsal carpal arch. AB - It is accepted that the ulnar artery forearm flap is dependent on the integrity of the palmar arches for ulnar arterial backflow and the patency of both radial and ulnar arteries. A patient is presented who demonstrates the successful use of the ulnar artery forearm flap supplied by the dorsal carpal arch via the dorsal branch of the ulnar artery. In this patient, the ulnar artery was damaged distal to the dorsal branch of the ulnar artery. A distally based ulnar forearm was dissected up to the dorsal branch of the ulnar artery and pivoted around this branch. This flap survived completely with no postoperative problem. PMID- 9746090 TI - Lateral supramalleolar flap for heel coverage in a patient with Werner's syndrome. AB - The repair of intractable ulcer over the Achilles' tendon in a patient with Werner's syndrome is described. The ulcer was covered successfully with a lateral supramalleolar flap combined with a delay procedure. Partial necrosis of the flap occurred; however, the necrotic area could be repaired with a skin graft because the underlying adipofascia of the flap survived. Histopathological studies revealed marked dermal fibrosis spreading to the subcutis, however the deep adipofascia was not so afflicted. The lateral supramalleolar flap, if combined with a delay procedure, can be used as a first choice in the repair of an ulcer in the heel region as long as apparent arteriosclerotic changes do not exist. PMID- 9746091 TI - Isolated Tessier no. 1 cleft of the nose. AB - We report 3 patients with isolated cleft or coloboma of the nose--an extremely rare occurrence. This entity belongs to no. 1 of Tessier's classification of craniofacial clefts. Excessive separation of the medial and lateral nasal processes of the frontonasal process may be the genesis of this condition. Management should address two points: the cleft and the associated malformations of the cartilaginous nasal framework. PMID- 9746092 TI - Malignant transformation of a giant proliferating trichilemmal tumor of the scalp: patient report and literature review. AB - Proliferating trichilemmal tumors are benign epitheliomas that may show malignant transformation. We present a trichilemmal tumor 15 x 15 x 10 cm in size and emphasize malignant transformation criteria. PMID- 9746093 TI - Fistula repair following antethoracic esophageal reconstruction using a radial forearm free flap. AB - The management of a fistula following retrosternal esophageal reconstruction proves particularly challenging. Two distinct tissue deficits must be considered for satisfactory reconstruction. The defect in the anterior wall of the colonic replacement and that of the adjacent soft tissues of the anterior chest wall can be closed simultaneously with a single, well-planned radial forearm free flap. A patient is presented who illustrates the design and implementation of a bivalved radial forearm free flap that provides two separate epithelial surfaces. PMID- 9746094 TI - Traumatic aneurysms of the face and temple: a patient report and literature review, 1644 to 1998. AB - The branches of the external carotid artery are protected from injury in most locations by an adequate buffer of soft tissue. On occasion, the vessels approach the surface to cross bone structures, and in these key areas they become vulnerable to blunt trauma. The facial, superficial temporal, and terminal branches of the internal maxillary arteries are the branches most often affected via this mechanism of injury. In addition, damage to deeper branches of the internal maxillary artery and to the subparotid portion of the superficial temporal artery has been reported secondary to maxillary fractures and craniofacial surgery. A brief patient report illustrates the highlights of clinical examination, diagnostic study, and surgical management of an aneurysm of the facial artery. A review of the world literature since 1644 has revealed 386 patients with traumatic aneurysms of the face and temple. PMID- 9746095 TI - An auricular paper model. AB - We have devised a paper model of the auricle that is simple to design and easily fabricated. An oval-shaped sheet of paper is folded so that the prominent and hollow parts center around a helical crus and concha. Most parts of the human auricle can be created by using this model. Studies of the model show that a large skin area is required for the helical crus. The model is considered to be useful in simulating total or partial auricular reconstruction. It also reveals new possibilities in reconstructive methods using tissues composed of cartilage and skin, which are flat and large. Furthermore, it is interesting that the formation of our paper model by forward rotation and folding is compatible with the process of auricle embryonic development. PMID- 9746096 TI - The ASPRS and reconstructive plastic surgery. American Society of Plastic and Reconstructive Surgeons. PMID- 9746097 TI - Re: Craniofacial injuries resulting from taxicab accidents in New York City. PMID- 9746098 TI - Re: Breast reconstruction after mastectomy without additional scarring: application of endoscopic latissimus dorsi muscle harvest. PMID- 9746099 TI - Rapid healing after skin laser resurfacing: a minimal mechanical trauma technique. PMID- 9746100 TI - Transparent hydrocolloid dressing for CO2 ultrapulse laser resurfacing. PMID- 9746101 TI - Hand burns in young patients--loss of self-sufficiency and economic productivity. PMID- 9746103 TI - Giant juvenile xanthogranuloma present since birth. PMID- 9746102 TI - Design-enhanced breast reduction: an approach for very large, very ptotic breasts without a vertical incision. PMID- 9746104 TI - A giant peduncular pilomatricoma. PMID- 9746105 TI - Skin flaps for pilonidal disease. PMID- 9746106 TI - Lipoma in the cheek of a 15-month-old child. PMID- 9746107 TI - Assessing the coronary circulation in hypertension. AB - Systemic arterial hypertension is one of the major risk factors for coronary artery disease, coronary microangiopathy, and left ventricular hypertrophy, all of which can potentially lead to cardiac failure and sudden cardiac death. Coronary flow reserve is defined as the maximal increase in coronary flow above its resting, autoregulated level for a given perfusion pressure. In arterial hypertension functional and structural alterations are observed at the level of epicardial vessels as well as in resistive vessels requiring sophisticated approaches to assess coronary flow reserve and thus myocardial perfusion. Electrocardiographic tests and echocardiography can be regarded as monitoring and screening methods. Myocardial scintography is useful to semiquantitatively estimate hypertension-associated perfusion abnormalities, whereas positron emission tomography provides the only quantitative approach of a non-invasive technique for myocardial blood flow measurement. Invasive methods for the assessment of coronary blood flow need cardiac catheterization procedures, such as techniques requiring catheterization of the coronary sinus, angiographic methods, and guidewire based methods. Thermodilution and venous oxymetry in the coronary sinus systematically underestimate coronary flow reserve and are thus considered as only semiquantitative approaches. In contrast, the gas chromatographic argon method allows a quantitative measurement of coronary blood flow at baseline and during maximum vasodilation; thus it is possible to distinguish between an altered autoregulated and maximal flow as the major cause of a reduced coronary flow reserve and to evaluate long-term therapeutic interventions in hypertensive hearts. Videodensitometric and angiographic methods should be restricted only to patients with coronary microangiopathy or with coronary single-vessel disease. Guidewire-based Doppler techniques are suitable to semiquantitatively assess coronary flow reserve with a considerable spatial and time resolution. Myocardial biopsies may gain insight into hypertension associated structural alterations in small arterioles. Long-term treatment of hypertensive heart disease aims to normalize blood pressure, to reduce left ventricular hypertrophy and to achieve cardioreparation including reversal of the abnormal structure and function of coronary circulation. Based on the different methods for assessment of coronary circulation the therapeutic value of different classes of antihypertensive therapeutics will be evaluated in this overview. PMID- 9746108 TI - Low diastolic blood pressure and mortality in a population-based cohort of 16913 hypertensive patients in North Karelia, Finland. AB - OBJECTIVE: To examine the relationship between mortality and diastolic blood pressure during treatment. DESIGN: A prospective follow-up of a population-based dynamic cohort of hypertensive patients. SETTING: Province of North Karelia, eastern Finland. PATIENTS: A cohort of 16 913 North Karelian hypertensive patients in hypertension register of the North Karelia Project, who had been followed up since 1972 until the end of 1985. Most of these patients had been under antihypertensive drug therapy during the follow-up. MAIN OUTCOME MEASURES: Death: all deaths (n = 4490), deaths from cardiovascular disease (n = 2995) and deaths from non-cardiovascular disease (n = 1495). RESULTS: Of all deaths, 17% of those among men and 24% of those among women were of patients with mean diastolic blood pressures less than 90 mmHg. We found a U-shaped relationship between diastolic blood pressure and total, cardiovascular and non-cardiovascular mortalities. We investigated this relationship in more detail using Cox regression model. Low diastolic blood pressure was associated independently with increased mortality for the patients aged 50 years or more at baseline. The occurrence of cardiac failure and other cardiovascular complications of hypertension were more important determinants of mortality than was low diastolic blood pressure alone. CONCLUSIONS: We demonstrated that there is an association between low diastolic blood pressure and mortality for treated hypertensive patients aged 50-69 years. The clinical importance of this relationship for patients without any cardiovascular complications of hypertension seems negligible. For patients with complications, these complications are likely to be primary factors causing greater than normal mortality and low diastolic blood pressure is mostly a secondary phenomenon. Our data do not lend support to speculations that there is overtreatment of hypertension, which would increase mortality through making diastolic blood pressures too low. PMID- 9746109 TI - Increased blood pressure reactivity in children of borderline hypertensive fathers. AB - OBJECTIVE: To evaluate the influence of heredity on blood pressure levels and reactivity in the offspring of borderline hypertensive and normotensive fathers. PARTICIPANTS AND OUTCOME MEASURES: Borderline hypertensive and normotensive men having normotensive wives (n = 25 and 26) were identified in a population screening program. Their children aged above 12 years were invited to participate. Seventeen having a borderline hypertensive father (BHT+) and 19 with a normotensive father (NT+) were investigated. Clinical and 24 h ambulatory blood pressure was measured, as well as blood pressure reactivity to an arithmetic mental stress test. RESULTS: The BHT+ group had a significantly higher clinical systolic blood pressure than the NT+ group (126 +/- 13 versus 115 +/- 7 mmHg, P< 0.01) but similar 24 h blood pressure levels. Systolic blood pressure variability (standard deviation of systolic blood pressure measurements each hour over 24 h) was significantly higher in the BHT+ group (18 +/- 4 versus 16 +/- 4 mmHg, P< 0.05). During mental stress test the BHT+ group had significantly higher increases in systolic and diastolic blood pressures at 4 min (NT+ 8% and 13% versus BHT+ 16% and 23% above baseline, P< 0.05) and significantly elevated DBP during the period after the stress test (NT+ 1% versus BHT+ 13% above baseline, P < 0.01). CONCLUSION: Even a mild level of hypertensive heredity affects important markers of blood pressure regulation, such as blood pressure variability and reactivity to mental stress. This might have prognostic implications; it also points to the possible importance of these variables as early signs of a familial predisposition to hypertension. PMID- 9746110 TI - Arterial hypertension due to occlusion of the adrenal vein in the rat is strain dependent. AB - OBJECTIVE: To determine whether the development of arterial hypertension due to occlusion of the central adrenal vein in the rat is strain-dependent DESIGN AND METHODS: The experiments were performed on male rats weighing 300-400 g each, of the following strains: Wistar outbred, Wistar Glaxo, Lewis, Wistar-Kyoto (WKY) rats bred for high blood pressure (138 +/- 13.2 mmHg), WKY rats bred without the control of blood pressure (118 +/- 12.9 mmHg) and borderline hypertensive rats (BHR). BHR were the F1 spontaneously hypertensive rat and WKY rat crossbred rats. In order to increase blood flow through the adrenal-renal portal circulation, both central adrenal veins of rats in the experimental group were occluded. The systolic blood pressure was measured indirectly by a photoelectric method. RESULTS: By the ninth day after surgery systolic blood pressure had increased significantly only in the WKY rats bred for high blood pressure and BHR, reaching maximal values on 12th day for WKY rats bred for high blood pressure (167 +/- 5 mmHg) and on the 18th day for BHR (170 +/- 14 mmHg). CONCLUSIONS: These data show that the development of arterial hypertension due to augmentation of adrenal blood flow through adrenal-renal portal circulation occurs in rats of strains with a genetic background of hypertension. PMID- 9746111 TI - Plasma levels of natriuretic peptides and adrenomedullin in elderly hypertensive patients: relationships to 24 h blood pressure. AB - OBJECTIVE: The aim of this study was to investigate the relationships between levels of natriuretic peptides and adrenomedullin and 24 h blood pressure levels in elderly hypertensives. DESIGN AND METHODS: We performed both 24 h ambulatory blood pressure monitoring and measurement of plasma levels of atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP) and adrenomedullin in 118 asymptomatic hypertensive elderly (> 60 years old) patients. We classified the subjects into groups with isolated clinic hypertension (n = 40) and sustained hypertension (n = 78). We also measured the levels of these peptides in 37 elderly normotensive subjects. RESULTS: Plasma ANP and BNP levels were slightly increased in patients with isolated clinic hypertension compared with elderly normotensives. Among the hypertensives, plasma ANP and BNP levels were more closely related to 24 h blood pressure levels than to office blood pressure levels. Sustained hypertensives showed significantly increased plasma levels of ANP and BNP compared with isolated clinic hypertensives, while adrenomedullin levels were similar in the two groups. Elderly hypertensives with left ventricular hypertrophy detected by electrocardiography had significantly higher levels of ANP and BNP, and higher BNP/ANP ratios than those without left ventricular hypertrophy, while there was no significant difference in adrenomedullin levels between the two groups. CONCLUSIONS: Our results suggest that measurements of ANP and BNP may be useful in detecting left ventricular hypertrophy and in differentiating isolated clinic hypertension from sustained hypertension in elderly hypertensive patients. PMID- 9746112 TI - Protein carboxyl methylation controls intracellular pH in human platelets. AB - OBJECTIVES: Carboxyl methylation is a reversible post-translational event which regulates the function of several cellular proteins. Because the human Na+-H+ antiporter (NHE-1) possesses a C-terminal consensus sequence for carboxyl methylation, we examined the role of protein carboxyl methylation in the regulation of intracellular pH homeostasis. DESIGN: Experiments were conducted using human platelets and N-acetyl-S-trans,trans-farnesyl-L cysteine (AFC), a specific prenylcysteine methyltransferase inhibitor. The effect of AFC on both basal intracellular pH (pHi) and on the kinetic properties of the Na+-H+ antiporter was characterized. MATERIALS AND METHODS: pHi was determined in cell suspensions using 2,7-biscarboxyethyl-5(6)-carboxyfluorescein tetraacetoxymethyl ester, a fluorescent pH indicator. The kinetics properties of the Na+-H+ antiporter activity were determined using platelets acidified with nigericin and challenged with varying extracellular concentrations of Na+. RESULTS: AFC (20 micromol/l) decreased basal pHi significantly (7.047 +/- 0.011 versus 7.133 +/- 0.012 for control, P< 0.001). The acidification was dose-dependent and reached steady state 3 min after AFC addition. In the absence of extracellular Na+, the platelets were acidified to the same extent with AFC or with ethanol (control): 6.530 +/- 0.031 versus 6.532 +/- 0.031 (P= 0.97). However, upon addition of Na+, the platelets treated with AFC showed a significant decrease in the maximal value for initial pHi recovery compared with controls: 0.788 +/- 0.041 versus 0.983 +/- 0.047 pH/min (P< 0.02). AFC also increased the Hill coefficient (2.89 +/- 0.22 versus 2.14 +/- 0.16, P < 0.03), and tended to decrease K0.5, the [Na+] corresponding to half-maximal activation (51.3 +/- 1.8 versus 60.5 +/- 3.9 mmol/l, P = 0.06) of the antiporter. CONCLUSION: Our data indicate that inhibition of carboxyl methylation reduces basal pHi and alters the kinetic properties of the Na+-H+ antiporter in human platelets, suggesting that carboxyl methylation is implicated in the regulation of intracellular pH homeostasis. PMID- 9746113 TI - Anti-oxidant status and lipid peroxidation in patients with essential hypertension. AB - BACKGROUND: Lipid peroxidation and derived oxidized products are being intensively investigated, because of their potential to cause injury and their pathogenetic role in several clinically significant diseases. The view that an excess of lipid peroxidation products is present and relevant in the pathogenesis of human essential hypertension or in hypertension-induced damage has still not received definitive support. OBJECTIVE: To evaluate both the extent of lipoperoxidation in essential hypertensive patients and the status of enzymatic and non-enzymatic antioxidants that potentially are able to modulate it METHODS: We selected 105 newly diagnosed essential hypertensives among those referred to our hypertension outpatient clinic and compared them with 100 normotensive controls matched for age. Plasma malondialdehyde was measured by high-performance liquid chromatography after reaction with thiobarbituric acid, as an end product of lipid peroxidation; serum selenium (Se), plasma copper (Cu) and zinc (Zn), vitamins A and E, erythrocyte superoxide dismutase and glutathione peroxidase levels were evaluated as indices of oxidant balance. Differences between the groups were tested by Student's t test and chi2 test. RESULTS: Compared with controls, essential hypertension patients had higher malondialdehyde and glutathione peroxidase activities (P<0.05 for both) and Zn concentrations (P<0.001) and lower superoxide dismutase activities (P<0.005), vitamin A (P<0.05) and E (P<0.001) levels and Cu concentrations (P<0.005). We found no difference between Se levels of essential hypertensive and control subjects. CONCLUSIONS: Essential hypertension is associated with greater than normal lipoperoxidation and an imbalance in anti-oxidant status, suggesting that oxidative stress is important in the pathogenesis of essential hypertension or in arterial damage related to essential hypertension. PMID- 9746114 TI - Cardiovascular kinin-generating capability in hypertensive fructose-fed rats. AB - OBJECTIVE: The hypertensive state is often associated with metabolic abnormalities, including glucose intolerance. Tissue kallikrein, a potent kinin generating enzyme, is present in the vascular wall and heart tissue. High dietary fructose consumption is reported to induce hyperinsulinemia, hypertriglyceridemia and hypertension. The objective of the present study was to examine the status of kallikrein in vascular and cardiac tissue from highly fructose-fed rats and to delineate the effect of kinins and the angiotensin converting enzyme inhibitor ramipril in this animal model of glucose intolerance. DESIGN AND METHODS: Male Wistar rats (350 g body weight) were divided into four groups of 10 rats each: (1) controls; (2) oral ramipril at 500 microg/kg per day for the last 2 study weeks; (3) fructose in drinking water as a 10% (w/v) solution for 4 weeks; and (4) fructose + ramipril, with fructose administered as in group 3 plus the administration of ramipril for the last 2 study weeks. Systolic blood pressure (tail-cuff method), glucose tolerance (2 g/kg body weight intraperitoneally) and metabolic parameters were recorded. Kallikrein activity in tail artery and heart tissue homogenates was estimated at the end of the 4th study week from measurements of kininogenase activity and kinins generated by a radioimmunoassay. RESULTS: The area under the curve for the glucose tolerance test increased from 1265 +/- 103 mmol/l after 120 min in the control and 1152 +/- 36 mmol/l in the ramipril group (NS) to 2628 +/- 143 mmol/l in the fructose group (P<0.01). The administration of ramipril to fructose-treated rats in group 4 improved glucose tolerance (2160 +/- 100 mmol/l; P<0.05 versus group 3). Blood pressure increased significantly in fructose-fed rats but fell markedly in fructose-fed rats treated with ramipril (P<0.01). Kallikrein activity measured in the heart and vessels increased as a consequence of fructose administration (P<0.05), but the administration of ramipril increased this parameter to a much greater extent (P<0.01 versus control group), which correlated closely with the decrease in blood pressure and the improvement in glucose tolerance observed in the fructose + ramipril group. CONCLUSIONS: The administration of fructose as a solution in the drinking water induced glucose intolerance and increased blood pressure. Treatment with the angiotensin converting enzyme inhibitor ramipril improved glucose tolerance and significantly diminished blood pressure. Cardiovascular kinin-generating capability increased in treated animals and this increase was even higher when rats were treated with ramipril, suggesting that kinins, acting as a paracrine hormonal system, can exert cardiovascular protection and contribute to the beneficial effects of angiotensin converting enzyme inhibitor. PMID- 9746115 TI - Endothelin-1 infusion inhibits plasma insulin responsiveness in normal men. AB - OBJECTIVES: Elevated plasma endothelin (ET)-1 levels have been described in insulin-resistant states such as syndrome X, obesity, non-insulin-dependent diabetes mellitus, and in some studies in essential hypertension. To investigate whether increases in circulating ET-1 to levels observed in insulin-resistant states can modulate insulin levels and/or insulin sensitivity in humans, we assessed these variables during low, non-pressor-dose ET-1 compared with placebo infusion. DESIGN: In a randomized, single blind, crossover design, 10 lean normotensive male subjects received either an intravenous infusion of subpressor doses of ET-1 dissolved in polygeline or a control infusion of polygeline only (placebo). Using dynamic assessment by the minimal model approach with the modified frequent sampling intravenous glucose tolerance test (FSIGT) the following and other parameters were measured: insulin sensitivity; acute insulin response to glucose (AIR(G)) calculated as the average of the three peak values between 2 and 5 min after injection of glucose from which the basal insulin levels were subtracted; the initial area under the curve (AUC(1-19)) from insulin values between time 0 and 19 min and the first-phase insulin secretion (phi1) from insulin kinetics parameters. RESULTS: ET-1 infusion reduced AIR(G) (to 34.85 +/- 4.27 compared with 49.3 +/- 6.9 microU/ml during placebo, P=0.017) and the acute C-peptide response to glucose (to 2.33 +/- 0.41 compared with 3.1 +/- 0.44 ng/ml, P=0.018), decreased plasma insulin levels during the FSIGT compared with placebo (analysis of variance P<0.0001) and decreased the AUC(1-19) (to 2.1 +/- 0.2 compared with 2.9 +/- 0.3 U/l per 20 min, P<0.01) while phi1 tended to be lower. S1 measured during ET-1 infusion was unaltered (11.11 +/- 1.91 x 10(-4) versus 10.88 +/- 2.11 10(-4)/min per mU per l, NS). CONCLUSIONS: These findings demonstrate that an increase in circulating ET-1 to levels observed in insulin resistant states acutely diminishes the insulin secretory response but does not significantly modify insulin sensitivity. PMID- 9746116 TI - Identical hemodynamic and hormonal responses to 14-day infusions of renin or angiotensin II in conscious rats. AB - OBJECTIVE: To investigate whether plasma angiotensin II (Ang II) determines the effects of the renin-angiotensin system or whether tissue uptake of renin and localized production of Ang II might account for any cardiovascular, renal, hormonal or drinking effect of circulating renin. DESIGN: Intravenous infusions of renin (0.6 ng/min; n = 10) and Ang II (3.5 ng/min; n = 10) that produce similar plasma Ang II levels were compared for 2 weeks with vehicle (n = 7) in conscious rats after a 1-week control period. Mean arterial pressure (MAP) and the heart rate were measured continuously. Hormones and renal function were measured twice weekly. Plasma Ang II and recovery data were measured in seven additional rats. RESULTS: In renin- and Ang II-infused rats, respectively, plasma Ang II increased similarly from 4.5 +/- 0.8 and 4.4 +/- 0.9 to 10.8 +/- 0.7 and 10.6 +/- 0.7 pg/ml and declined similarly in the second week to 7.0 +/- 1.1 and 7.0 +/- 1.5 pg/ml. Plasma renin increased from 4.2 +/- 0.7 to 21.7 +/- 1.3 and fell from 5.9 +/- 0.5 to 0.6 +/- 0.2 ng/ml per h respectively. Plasma prorenin fell similarly (> 70%); angiotensinogen was unchanged. MAP rose initially by 25.6 +/- 1.2 and 23.3 +/- 0.9 mmHg and by an additional 21.1 +/- 2.4 and 27.4 +/- 1.8 mmHg on days 5-8. The heart rate fell gradually but transiently by -11% in both. Although the initial MAP rise was slower in renin-infused rats (P< 0.05) MAP returned to baseline within 2 h after both infusions were stopped. Changes in renal vascular resistance, renal blood flow, glomerular filtration rate, urinary sodium, potassium and water excretion and water intake were not significantly different between renin- and Ang II-infused rats. CONCLUSIONS: Intravenous infusions of low doses of renin or Ang II into conscious rats increase MAP identically. MAP increases in two phases 5-8 days apart, in coordination with transient falls in the heart rate. Renin- and Ang II-induced chronic hypertension are identically sustained by very small increases in plasma Ang II. Blood pressure increases more slowly with renin infusions, consistent with tissue binding. Notwithstanding, no evidence was obtained for a physiological role of tissue-bound renin in causing the cardiovascular, renal, hormonal and drinking responses measured in this study. PMID- 9746117 TI - Effects of angiotensin II and losartan in the forearm of patients with essential hypertension. AB - OBJECTIVE: Angiotensin II type 1 receptor-mediated constrictor effects may be modulated by hypertension-related vascular changes, changes in receptor function and in neurohumoral activity. DESIGN: The forearm blood flow (FBF) effects of angiotensin II, methoxamine, and losartan were investigated in essential hypertensive patients. Minimal forearm vascular resistance was measured to determine structural vascular changes. METHODS: Seven hypertensive patients were selected, and seven matched normotensives. Angiotensin II (0.01-10 ng/kg per min) was infused during predilatation by sodium nitroprusside (6.1 +/- 0.6 ng/kg per min) before and during losartan infusion (0.3-3 microg/kg per min). Methoxamine (0.2-2 microg/kg per min) was co-infused with the nitric oxide synthase inhibitor NG-monomethyl-L-arginine. FBF, measured by venous occlusion plethysmography, was expressed as the change in FBF ratio (FBFinfused arm/FBFnon-infused arm). RESULTS: Baseline FBF (infused arm) was increased by sodium nitroprusside from 2.56 +/- 0.80 to 5.46 +/- 0.92 (P<0.05) and from 2.66 +/- 0.25 to 5.42 +/- 0.40 ml/100 ml per min (P<0.05) in the hypertensive and normotensive group, respectively. Baseline forearm vascular resistance (FVR) was higher in the hypertensive than in the normotensive group [51 +/- 8 versus 33 +/- 3 mmHg/ (ml/100 ml per min); P<0.05]. Angiotensin II caused a maximal change in FBF ratio (Emax) by -70 +/- 3 and -72 +/- 6% in the hypertensive and normotensive group, respectively (NS). Tachyphylaxis did not occur. Infusions of losartan at 0.3, 1 and 3 microg/kg per min reduced the Emax values from -70 +/- 3 to -50 +/- 5, -45 +/- 5 and -15 +/- 2%, respectively, in the hypertensive group, and from -72 +/- 6 to -62 +/- 4, -45 +/- 2 and -32 +/- 2%, respectively, in the normotensive group (NS). Infusion of methoxamine significantly reduced the FBF ratio by -58 +/- 6 and -69 +/- 5% in the hypertensive and normotensive groups, respectively (NS). Minimal FVR, after forearm ischemia, was the same in hypertensives and normotensives, namely 3.2 +/- 0.7 and 3.2 +/- 0.4 mmHg/(ml per 100 ml per min), respectively (NS). CONCLUSIONS: Angiotensin II type 1- and alpha1-mediated vascular effects were unchanged by essential hypertension. Baseline FVR was greater in the hypertensives than in the normotensives, while minimal FVR was the same. These results indicate that the forearm vascular bed of the patient group studied does not show important structural and renin-angiotensin system-related functional changes as a result of hypertension. PMID- 9746118 TI - Alteration of association of agonist-activated renal D1(A) dopamine receptors with G proteins in proximal tubules of the spontaneously hypertensive rat. AB - BACKGROUND: Defective D1A dopamine receptor-G protein coupling has been identified in renal proximal tubules of the spontaneously hypertensive rat (SHR). OBJECTIVE: To determine whether association of D1A dopamine receptors with the alpha subunits of G proteins in kidney of SHR is normal. METHODS: We analyzed the association of agonist-activated [1251]-labeled D1A dopamine receptors in kidneys of SHR and the normotensive Wistar-Kyoto (WKY) rat through immunoprecipitation, using highly specific antipeptide antibodies directed against alpha subunits of G proteins. RESULTS: We have shown for the first time that the D1A receptors of renal proximal tubules are associated with the adenylyl cyclase inhibitory G proteins G(i)alpha. The association of WKY rat proximal tubule D1A receptors with Gi1alpha and Gi2alpha in the presence of agonist is significantly (P<0.01) greater (2.4-fold and 3.1-fold greater, respectively) than it is without agonist D1A receptors of WKY rat also exhibit (twofold greater) association with G(s)alpha, consistently with the ability of these receptors to mediate stimulation of adenylyl cyclase. The WKY rat D1A receptors do not associate either with G(o)alpha or with G(q)alpha. The D1A receptors of SHR proximal tubule membranes appear to be resistant to activation by agonist and do not associate with G(s)alpha, G(o)alpha and any of the subunits of G(i)alpha. However, the SHR D1A sites exhibit a modestly (1.7-fold) greater association with G(q)alpha, which was not statistically significant. The differences among associations of the D1A receptors of WKY rat and SHR with these Galpha proteins may be important in understanding renal dopaminergic functions in normal and pathophysiologic states. PMID- 9746119 TI - Isoform-specific regulation of nitric oxide synthase mRNA in the kidney by sodium and blood pressure. AB - BACKGROUND: The roles of nitric oxide synthases (NOS) in kidney function are still controversial, principally due to the lack of isoform-specific inhibitors of NOS. OBJECTIVE: To investigate the relative roles of each isoform of NOS in regulation of sodium and volume homeostasis. DESIGN: We studied the effects of long-term modifications of sodium diet and blood pressure on expression of NOS mRNA in the renal cortex, where the three isoforms of NOS are present. METHODS: We used quantitative reverse-transcription-polymerase chain reaction assays specific to each isoform of NOS to determine amounts of their respective mRNA in control rats, deoxycorticosterone acetate (DOCA)-salt hypertensive rats, rats fed a high-salt diet, and furosemide-treated rats fed a low-sodium diet. Nicotinamide adenine nucleotide phosphate H (NADPH) diaphorase staining was performed on DOCA salt and control rat kidneys. RESULTS: Levels of NOS I mRNA in DOCA-salt rats were decreased by treatment, those in low-salt-diet rats remained unaffected and those in high-salt diet rats tended to be intermediate between those of the other rat groups. Expression of NOS III mRNA was not significantly modified by either treatment Levels of NOS II mRNA in DOCA-salt rats were increased, those in high salt-diet rats remained unaffected, and those in low-salt-diet were decreased by treatment, but these levels are more than 100-fold lower than those observed for the other isoforms of NOS. NADPH diaphorase staining in macula densa of DOCA-salt rats was markedly decreased compared with that in macula densa of control rats but staining in renal inflammatory and fibrous lesions became detectable, and staining in the vessels did not differ from that for control rats. CONCLUSIONS: Our results show that intake of sodium and extracellular fluid volume regulate levels of mRNA of the three NOS isoforms in the renal cortex differently, suggesting that each of them plays a specific role. PMID- 9746120 TI - Microalbuminuria predicts cardiovascular events and renal insufficiency in patients with essential hypertension. AB - BACKGROUND: Some patients with essential hypertension manifest greater than normal urinary excretion of albumin (UAE). Authors of a few retrospective studies have suggested that there is an association between microalbuminuria and cardiovascular risk. OBJECTIVE: To evaluate whether microalbuminuria is associated with a greater than normal risk of cardiovascular and renal events. METHODS: We performed a retrospective cohort analysis of 141 hypertensive individuals followed up for approximately 7 years. Hypertensive patients were defined as having microalbuminuria if the baseline average UAE of three urine collections was in the range 30-300 mg/24 h. RESULTS: Fifty-four patients had microalbuminuria and 87 had normal UAE. At baseline, the two groups were similar for age, weight, blood pressure, and rate of clearance of creatinine. Serum levels of cholesterol, triglycerides, and uric acid in patients with microalbuminuria were higher than levels in those with normal UAE, whereas levels of high-density lipoprotein cholesterol in patients with microalbuminuria were lower than levels in patient with normal UAE. During follow-up, 12 cardiovascular events occurred among the 54 (21.3%) patients with microalbuminuria and only two such events among the 87 patients with normal UAE (P < 0.0002). Stepwise logistic regression analysis showed that UAE (P = 0.003), cholesterol level (P = 0.047) and diastolic blood pressure (P = 0.03) were independent predictors of the cardiovascular outcome. Rate of clearance of creatinine from patients with microalbuminuria decreased more than did that from those with normal UAE (decrease of 12.1 +/- 2.77 versus 7.1 +/- 0.88 ml/min, P < 0.03). CONCLUSIONS: This study suggests that hypertensive individuals with microalbuminuria manifest a greater incidence of cardiovascular events and a greater decline in renal function than do patients with normal UAE. PMID- 9746121 TI - Adverse prognostic value of a blunted circadian rhythm of heart rate in essential hypertension. AB - BACKGROUND: Previous studies revealed a direct association between resting heart rate and risk of mortality in essential hypertension. However, resting heart rate is a highly variable measure since it is affected by the alerting reaction to the visit. OBJECTIVE: To investigate whether the heart rate values recorded during the 24 h of ambulatory blood pressure monitoring are independent predictors of survival of uncomplicated subjects with essential hypertension. METHODS: We followed up 1942 initially untreated and uncomplicated subjects with essential hypertension (mean age 51.7 years, 52% men) for an average of 3.6 years (range 0 10 years). All subjects underwent baseline procedures including 24 h non-invasive blood pressure monitoring with simultaneous assessment of heart rate, one reading every 15 min for 24 h. MAIN OUTCOME MEASURES: All-cause mortality and cardiovascular morbidity. RESULTS: During follow-up there were 74 deaths from all causes (1.06 per 100 person-years) and 182 total (fatal plus non-fatal) cardiovascular morbid events (2.66 per 100 person-years). Clinic, average 24 h, daytime and night-time heart rates exhibited no association with total mortality. However, the subjects who subsequently died had had a blunted reduction of heart rate on going from day to night during the baseline examination. After adjustment for age (P < 0.001), diabetes (P < 0.001) and average 24 h systolic blood pressure (SBP, P= 0.002) in a Cox model, for each 10% less reduction in the heart rate from day to night the relative risk of mortality was 1.30 (95% confidence interval 1.02-1.65, P = 0.04). Rates of death were 0.38, 0.71, 0.94 and 2.0 per 100 person-years among subjects in the four quartiles of the distribution of the percentage reduction in heart rate from day to night The baseline day-night changes in the heart rate exhibited an inverse correlation to age and to clinic and ambulatory SBP and a direct association with the day-night changes in blood pressure. The degree of reduction of heart rate from day to night also had an independent inverse association with total cardiovascular morbidity after adjustment for age, diabetes and left ventricular hypertrophy, but this association did not remain significant when average 24 h SBP and the degree of day-night reduction in SBP were entered into the equation. CONCLUSIONS: A flattened diurnal rhythm of heart rate in uncomplicated subjects with essential hypertension is a marker of risk for subsequent all-cause mortality and this association persists after adjustment for several risk factors. For assessing these subjects, a limited and uniformly distributed period of ambulatory heart rate recording during the 24 h is clinically valuable. PMID- 9746124 TI - Undertreatment of hypertension in a population-based study in The Netherlands. AB - OBJECTIVE: To estimate the level of undertreatment of hypertension in a population-based study by taking into account the co-existence of additional cardiovascular risk factors in untreated hypertensives, uncontrolled blood pressure among pharmacologically treated hypertensives and within-person variability in blood pressure and total cholesterol. DESIGN: Cross-sectional. SETTING: Two population-based surveys on cardiovascular disease risk factors conducted during 1987-1995 in The Netherlands. PARTICIPANTS: 56 026 men and women aged 20-59 years. MAIN OUTCOME MEASURES: Prevalence of hypertension, of treatment and of undertreatment of hypertension. Undertreated hypertensives were those who were treated pharmacologically, but whose blood pressure was still elevated and those who inappropriately received no medication for the treatment of hypertension. RESULTS: During the past decade in The Netherlands, 30% of the hypertensive women and 47% of the hypertensive men aged 20-59 years were undertreated for hypertension. In both men and women treated pharmacologically, 42 and 29%, respectively, still had elevated blood pressure levels. Of those hypertensive men and women not treated pharmacologically, 53 and 34%, respectively, should have been treated when additional cardiovascular risk factors were taken into account Among those diagnosed but untreated for hypertension, 58 and 31% of the men and women, respectively, should have been treated pharmacologically. CONCLUSION: A considerable proportion of hypertensives were undertreated for hypertension. To decrease the undertreatment of hypertension, it is necessary to obtain better control of blood pressure in patients already being treated, increase the detection of hypertension and improve adherence to the current guidelines. PMID- 9746122 TI - Regulation of extracellular matrix proteins in pressure-overload cardiac hypertrophy: effects of angiotensin converting enzyme inhibition. AB - OBJECTIVE: Left ventricular hypertrophy (LVH) is characterized by remodeling of both myocyte and interstitial compartments of the heart. The aim of this investigation was to study the effects of angiotensin converting enzyme (ACE) inhibition on alterations in the composition of the interstitium in chronic pressure-overload hypertrophy. DESIGN: LVH was induced in weanling rats by banding the ascending aorta. Animals with aortic banding received either vehicle (n = 20), hydralazine (20 mg/kg per day, n = 20), or the ACE inhibitor ramipril (10 mg/kg per day, n = 20) during weeks 6-12 after banding. RESULTS: Compared with sham-operated, untreated rats (n = 20), aortic-banded vehicle and hydralazine-treated rats displayed substantially increased left ventricular weights and myocyte diameters whereas ramipril significantly blunted the hypertrophic response at the myocyte level (each P < 0.001) as well as the increase in left ventricular weight (each P < 0.01). In addition, image analysis revealed a significant induction of perivascular and interstitial tissue accumulation in vehicle- and hydralazine-treated rats (2.5-fold, each P < 0.0001). In contrast, ramipril-treated rats displayed attenuated interstitial and perivascular fibrosis, both being significantly diminished compared with vehicle- and hydralazine-treated rats (each P< 0.001). Further, vehicle- and hydralazine treated rats were characterized by elevated steady-state messenger (m)RNA levels of fibronectin (2.7- and 2.8-fold, P< 0.005), collagen I (2.0- and 1.8-fold, P < 0.0005), collagen III (both 2.2-fold, P < 0.001) and laminin B (1.6- and 1.6 fold, P < 0.005). In parallel, the corresponding immunohistochemical signals were markedly enhanced in these groups. In comparison, ramipril significantly blunted the induction of collagen I and III, laminin B and fibronectin at both the mRNA and protein levels. These morphological and molecular differences between the hydralazine and ramipril groups could not be attributed to differences in left ventricular-pressures, which were markedly elevated in all aortic stenosis rats (1.9-fold, each P < 0.001 versus sham). In fact, given that ramipril but not hydralazine blunted the hypertrophic response to pressure overload, the echocardiographic measurements revealed that left ventricular systolic wall stress was higher in the ramipril group (70 +/- 1 versus 34 +/- 0.7 kdyn/cm2; P < 0.02). CONCLUSIONS: ACE inhibition may limit both myocyte and interstitial remodeling despite ongoing cardiac pressure overload. PMID- 9746123 TI - Different effects of nifedipine and amlodipine on circulating catecholamine levels in essential hypertensive patients. AB - OBJECTIVE: To compare the acute and chronic effects of nifedipine retard (NPA), nifedipine gastrointestinal therapeutic system (NGITS) and amlodipine at trough and peak plasma concentrations of drug on blood pressure and heart rate, and on plasma norepinephrine and epinephrine levels in patients with mild-to-moderate hypertension (diastolic blood pressure 95-115 mmHg). DESIGN AND METHODS: After 3 4 weeks' placebo treatment, patients of both sexes were randomly allocated to be administered 10 or 20 mg NPA twice a day, 30 or 60 mg NGITS once a day, and 5 or 10 mg amlodipine once a day for 6 weeks. Initially, for the first 2 weeks, the lowest dose of each drug was used, but higher doses were administered after 2 weeks if sitting diastolic blood pressure was > 90 mmHg. Patients were evaluated after administration of the first dose and after 6 weeks' therapy in a hospital setting. Blood samples were taken for high-performance liquid chromatography measurement of catecholamine and drug levels at various intervals for a period covering trough to peak drug level ranges. RESULTS: Administration of all three drugs reduced clinic blood pressure to the same level after 6 weeks' therapy, but heart rate was increased slightly only with amlodipine (P < 0.05). Administration of NPA reduced blood pressure more abruptly whereas administrations of NGITS and amlodipine induced smoother falls after acute and chronic treatments: a significant increase in heart rate was observed with amlodipine after chronic treatment. Both acute and chronic treatments with NPA (n = 19) increased norepinephrine levels (P < 0.01) transiently (2-4 h). In contrast, administration of NGITS (n = 22) did not increase norepinephrine levels and even induced a slight but significant decrease in norepinephrine levels 5-6 h after chronic treatments. Although administration of amlodipine (n = 22) did not increase norepinephrine levels transiently either after acute or after chronic administration, it did induce a sustained rise in basal norepinephrine levels by more than 50% after chronic therapy (P < 0.01). Plasma epinephrine levels were not increased by any of the treatments and even a slight decrease was observed 4 h after administration of a dose following chronic treatments with NGITS and amlodipine (P < 0.05). CONCLUSIONS: The transient increase in norepinephrine levels observed with NPA and the sustained increases in norepinephrine levels observed after chronic treatment with amlodipine suggest that sympathetic activation occurs with those two drugs. The lack of increase in norepinephrine levels after administration of NGITS suggests that this formulation does not activate the sympathetic system. The lowering of epinephrine levels after administrations of NGITS and amlodipine suggests that inhibition of release of epinephrine by the adrenal medulla occurs with longer-acting dihydropyridine formulations. PMID- 9746125 TI - Trends in blood pressure levels and control of hypertension in Finland from 1982 to 1997. AB - OBJECTIVE: To assess the trends in blood pressure levels and hypertension control in Finland from 1982 to 1997. DESIGN: Four independent cross-sectional population surveys conducted in 1982, 1987, 1992 and 1997. SETTING: From 1982 to 1997, the provinces of North Karelia and Kuopio in eastern Finland and the region of Turku Loimaa in southwestern Finland were surveyed. From 1992 to 1997, the Helsinki Vantaa region in southern Finland was surveyed. PARTICIPANTS: Men and women aged 25-64 years were selected randomly from the national population register. The total number of participants was 27 623. MAIN OUTCOME MEASURES: We assessed mean systolic and diastolic blood pressure, prevalence of hypertension (subjects with systolic blood pressure > or = 160 mmHg or diastolic blood pressure > or = 95 mmHg or current use of antihypertensive drug treatment) and antihypertensive drug treatment and quality of hypertension care among hypertensive persons. RESULTS: Mean systolic blood pressure and the prevalence of hypertension decreased significantly in all areas except among men in the Helsinki-Vantaa region. The fall in mean diastolic pressure was significant only in eastern Finland. The proportion of hypertensives who were unaware of their condition fell from 45.5 to 24.1% in men and from 27.2 to 15.7% in women. At the same time, the proportion of hypertensives with adequately controlled blood pressure (systolic pressure < 160 mmHg and diastolic pressure < 95 mmHg) increased from 9.4 to 23.5% in men and from 16.0 to 36.7% in women. CONCLUSION: Hypertension care in Finland has improved significantly during the last 15 years. However, the situation is still far from optimal. It is obvious that the biggest problem in hypertension care has shifted from detection to adequate treatment of high blood pressure. PMID- 9746126 TI - Atherosclerosis and left ventricular hypertrophy: persisting problems in treated hypertensive patients. AB - OBJECTIVES: First, to determine whether hypertensive patients managed in general practice have more advanced atherosclerosis and left ventricular hypertrophy than matched normotensive patients from the same practices. Second, to investigate the associations of several potentially modifiable factors with these vascular and cardiac outcomes. DESIGN AND METHODS: We performed a case-control study of 500 hypertensive cases (systolic blood pressure > or = 160 mmHg or diastolic blood pressure > or = 95 mmHg or receiving treatment) and 506 age- (mean 61 years) and sex- (54% female) matched normotensive controls recruited from general practices. Carotid artery far wall thickness (CWT), assessed by B-mode ultrasound, and left ventricular mass (LVM), assessed by M-mode echocardiography, were the main study outcome measures. RESULTS: Mean systolic/diastolic blood pressure levels in the 399 treated cases (145/87 mmHg) were lower than those in untreated cases (158/94 mmHg) but higher than those in controls (133/82 mmHg, all P < 0.0001). Mean body mass index (BMI) and total triglyceride levels were higher and high-density lipoprotein cholesterol was lower in cases than in controls (all P < 0.0004). Mean CWT was 10% greater in cases than in controls and LVM was 14% greater (both P < 0.001), but both were similar in treated and untreated cases (P > 0.05). In multivariate analyses, blood pressure and BMI were both directly and independently related to CWT and LVM (both P < 0.0001). CONCLUSIONS: In this study, hypertensive patients managed in general practice - whether treated with antihypertensive drugs or not - had more advanced atherosclerosis and left ventricular hypertrophy than did matched normotensive patients. Efforts to lower blood pressure further and to reduce BMI could potentially reduce these differences, and this might lead to a reduction in the risk of major cardiovascular events. PMID- 9746127 TI - Mercury manometers and mmHg should be preserved. PMID- 9746128 TI - Load compensation during homologous and non-homologous coordination. AB - Two-limb coordination of homologous and non-homologous effectors was examined during isofrequency (1:1) and multifrequency (2:1) conditions. The coordination patterns involved flexion and extension movements in the sagittal plane and were performed under unloaded and single-limb (right arm) loaded conditions. Previous studies suggested that the lower degree of 1:1 synchronization observed during nonhomologous as compared to homologous coordination results from natural differences in biophysical (inertial) properties. Elaborating on this idea, adding weight to the right arm was hypothesized to modulate its inertial characteristics, rendering homologous limbs more dissimilar and nonhomologous limbs more similar by enhancing and decreasing their inertial differences, respectively. Therefore, the observations made during unloaded conditions were predicted to be completely reversed during loaded conditions. Findings revealed that during 1:1 coordination (experiment 1) single-limb loading resulted in a decreased relative phase stability, whereas relative phase accuracy depended upon the limb combination. In particular, phase-locking was more accurately maintained for loaded homologous than for nonhomologous limbs, whereas loading the nonhomologous limbs resulted in a deterioration of the quality of synchronization. These findings suggest that there is an additional explanation of differential coordination capabilities among limb combinations. It is hypothesized that the neural networks subserving the control centers of the homologous limbs are more tightly connected than those of the nonhomologous effectors, allowing 1:1 synchronization to be more successfully preserved in the face of (load) perturbations. During 2:1 coordination (experiment 2), the loading procedure disturbed the coordination dynamics across all limb combinations. That no differential effect of loading on effector combination was observed is possibly a result of the fact that only an initial level of practice was studied in which optimal relative phase dynamics are still being explored for both homologous and nonhomologous limbs. PMID- 9746129 TI - Influence of head-to-trunk position on sound lateralization. AB - The effect of horizontal head position on the lateralization of dichotic sound stimuli was investigated in four experiments. In experiment 1, subjects adjusted the interaural level difference (ILD) of a stimulus (band-pass noise) to the subjective auditory median plane (SAMP) while simultaneously directing the beam of a laser attached to the head to visual targets in various directions. The adjustments were significantly correlated with head position, shifting in a direction toward the side to which the head was turned. This result was replicated in experiment 2, which employed a two-alternative forced-choice method, in which stimuli of different ILD were presented and left/right judgments were made. In both experiments, the average magnitude of the shift of the SAMP was about 1 dB over the range of head positions from straight ahead to 60 degrees to the side. The shift of the SAMP indicates that any shift in head position induces a change in sound lateralization in the opposite direction, i.e., the intracranial sound image is shifted slightly to the left when the head is directed to the right and to the right when the head is to the left. In experiments 3 and 4, the effect of head position was compared with that of eye position by using the same methods as in experiment 2. Both shifts in SAMP, induced by either head- or eye-position changes, are in the same direction and, on average, of about the same magnitude (experiment 3), and head- and eye position effects compensate approximately for each other during variations of head position when the gaze remains fixed to a visual target in space (experiment 4). PMID- 9746130 TI - Hierarchical control of different elbow-wrist coordination patterns. AB - The present paper focused on the role of mechanical factors arising from the multijoint structure of the musculoskeletal system and their use in the control of different patterns of cyclical elbow-wrist movements. Across five levels of cycling frequency (from 0.45 Hz up to 3.05 Hz), three movement patterns were analyzed: (1) unidirectional, including rotations at the elbow and wrist in the same direction; (2) bidirectional, with rotation at the joints in opposite directions, and (3) free-wrist pattern, which is characterized by alternating flexions and extensions at the elbow with the wrist relaxed. Angular position of both joints and electromyographic activity of biceps, triceps, the wrist flexor, and the wrist extensor were recorded. It was demonstrated that control at the elbow was principally different from control at the wrist. Elbow control in all three patterns was similar to that typically observed during single-joint movements: elbow accelerations-decelerations resulted from alternating activity of the elbow flexor and extensor and were largely independent of wrist motion at all frequency plateaus. The elbow muscles were responsible not only for the elbow movement, but also for the generation of interactive torques that played an important role in wrist control. There were two types of interactive torques exerted at the wrist: inertial torque arising from elbow motion and restraining torque arising from physical limits imposed on wrist rotation. These interactive torques were the primary source of wrist motion, whereas the main function of wrist-muscle activity was to intervene with the interactive effects and to adjust the wrist movement to comply with the required coordination pattern. The unidirectional pattern was more in agreement with interactive effects than the bidirectional pattern, thus causing their differential difficulty at moderate cycle frequencies. When cycling frequency was further increased, both the unidirectional and bidirectional movements lost their individual features and acquired features of the free-wrist pattern. The deterioration of the controlled patterns at high cycling frequencies suggests a crucial role for proprioceptive information in wrist control. These results are supportive of a hierarchical organization of control with respect to elbow-wrist coordination, during which the functions of control at the elbow and wrist are principally different: the elbow muscles generate movement of the whole linkage and the wrist muscles produce corrections of the movement necessary to fulfill the task. PMID- 9746131 TI - Development of postural control of gravity forces in children during the first 5 years of walking. AB - The aim of this work was to propose developmental indexes relative to the control of balance and gravity forces, using force-plate data, for children in their first 5 years of independent walking. The first part of this paper is devoted to the definition of an index to quantify postural capacity during walking. Based on the assumption that the vertical acceleration of center of mass (CM) reflects the capacity of muscular forces between the head-arms-trunk and the stance leg segments to control the external forces, the value of the CM vertical acceleration at foot contact is proposed as a developmental index of the postural capacity of the child to control gravitational forces. This index was analyzed longitudinally in five children, over the course of eight experimental sessions. The children were examined during their first 5 years of independent walking (for a total of 457 step sequences). The covariation between the CM vertical acceleration at foot contact and the gait velocity was considered as a second index characterizing the development of coordination between the postural and dynamic requirements of body progression. From these indexes it was established that the postural capacity needed just to control balance with the leg muscles was not attained before 4-5 years of independent walking, i.e., at about 5-6 years of age. PMID- 9746132 TI - Tactile impairments cannot explain the effect of age on a grasp and lift task. AB - This experiment addressed the often-posed theory that age-related declines in manual dexterity result from diminished tactile function. We measured the time 'young' subjects (n=33; mean=45 years) and 'old' subjects (n=33; mean=74 years) needed to grip (thumb and index finger), lift, and transport a small metal sphere when vision was permitted and when blindfolded. Subjects began each trial by reaching for the sphere and were instructed to complete the entire task quickly. In the absence of visual information, placement of the finger and thumb for a secure grip and lift cannot be performed efficiently without tactile information. If age-related tactile changes are functionally significant for this task, then without visual information the 'old' group should show a disproportionate increase in the duration of the grip and lift phase of the task compared to the 'young' group. Perceptual thresholds for tactile pressure stimuli (Semmes Weinstein filaments) confirmed well-known age-related changes. Age and vision effects were manifest mainly during the grip-lift phase (time from object contact to lift-off from its support surface), with the expected finding that the 'old' group required more time than 'young' group, regardless of visual condition. The main finding was that the 'grip-lift' duration in the 'no-vision' condition was about twice the duration observed in the 'vision' condition for both age groups (ratios of 2.1 and 2.3 for 'young' and 'old', respectively). This similar relative slowing for the two groups fails to support the hypothesis that old adults' ability to grip and lift the object was limited by changes in the availability or use of tactile information. PMID- 9746133 TI - The timing of color and location processing in the motor context. AB - In this study, the use of color and location as stimulus attributes manipulated during a simple action was aimed at comparing how dorsal (location) and ventral (color) features are integrated in action and the timing of their processing. Eighteen subjects were presented with a green dot on a computer screen, which they were required to point at and touch. In 20% of the trials, the location or the color of the target was altered at the onset of movement to this stimulus, requiring the participant to modify the initially programmed response according to specific motor instructions. In the 'location-go' group, the target changed in location and participants were instructed to reach the displaced stimulus by correcting their ongoing movement. In the 'location-stop' and 'color-stop' groups, subjects were instructed to interrupt their movement when the target changed location or color, respectively. Results showed that the latency of the first responses to the perturbation clearly depended on the stimulus attribute and not on the motor instruction tested: the response to color change was obtained about 80 ms later than both conditions involving location change. It is concluded that: (1) color processing is slower than location processing, and (2) the first reactions to the location change occur after the same delay irrespective of the response required from the subject. PMID- 9746134 TI - Changes in the extracellular levels of glutamate and aspartate during ischemia and hypoglycemia. Effects of hypothermia. AB - Hypothermia (33 degrees C) dramatically diminishes ischemic but not hypoglycemic brain damage. The beneficial effects of hypothermia in ischemia have been partly attributed to a reduction in the ischemia-induced increase in synaptic levels of glutamate or aspartate. With the microdialysis technique, we studied the effects of hypothermia (33 degrees C) on the brain extracellular levels of glutamate and aspartate during hypoglycemia, ischemia, and their combination. In isoelectric hypoglycemia, striatal levels of glutamate and aspartate frequently show large transients of transmitter release occurring during both normothermia and hypothermia, whereas in the cortex levels of glutamate and aspartate are slightly lower during hypothermia compared with normothermia. In both regions studied, complete ischemia induced by i.v. KCl results in a progressive increase in glutamate and aspartate levels over time. In normoglycemic animals, hypothermia markedly attenuates the increase in glutamate and aspartate levels in the striatum but not in the cortex. Also in hypoglycemic animals, complete ischemia causes a progressive increase in the glutamate and aspartate levels. However, hypothermia affects only striatal glutamate levels. Since hypothermia protects both cortex and striatum against ischemic brain injury and not against hypoglycemic injury, presumably the protective effect of hypothermia is due to factors other than prevention of glutamate or aspartate overflow. PMID- 9746135 TI - Changes in motor cortical activity during visuomotor adaptation. AB - We examined neuronal activity in three motor cortical areas while a rhesus monkey adapted to novel visuomotor transforms. The monkey moved a joystick that controlled a cursor on a video screen. Each trial began with the joystick centered. Next, the cursor appeared in one of eight positions, arranged in a circle around a target stimulus at the center of the screen. To receive reinforcement, the monkey moved the joystick so that the cursor contacted the target continuously for Is. The video monitor provided continuous visual feedback of both cursor and target position. With those elements of the task constant, we modified the transform between joystick movement and that of the cursor at the beginning of a block of trials. Neuronal activity was studied as the monkey adapted to these novel joystick-cursor transforms. Some novel tasks included spatial transforms such as single-axis inversions, asymmetric double-axis inversions and angular deviations (also known as rotations). Other tasks involved changes in the spatiotemporal pattern and magnitude of joystick movement. As the monkey adapted to various visuomotor tasks, 209 task-related neurons (selected for stable background activity) showed significant changes in their task-related activity: 88 neurons in the primary motor cortex (M1), 32 in the supplementary motor cortex (M2), and 89 in the caudal part of the dorsal premotor cortex (PMdc). Slightly more than half of the sample in each area showed significant changes in the magnitude of activity modulation during adaptation, with the number of increases approximately equaling the number of decreases. These data support the prediction that changes in task-related neuronal activity can be observed in M1 during motor adaptation, but fail to support the hypothesis that M1 and PMdc differ in this regard. When viewed in population averages, motor cortex continued to change its activity for at least dozens of trials after performance reached a plateau. This slow, apparently continuing change in the pattern and magnitude of task-related activity may reflect the initial phases of consolidating the motor memory for preparing and executing visuomotor skills. PMID- 9746136 TI - Acute and long-lasting changes in extracellular-matrix chondroitin-sulphate proteoglycans induced by injection of chondroitinase ABC in the adult rat brain. AB - Lattice-like perineuronal accumulations of extracellular-matrix proteoglycans have been shown to develop during postnatal maturation and to persist throughout life as perineuronal nets (PNs) in many brain regions. However, the dynamics of their reorganization in adults are as yet unknown. The aim of the present study was to examine the capability of PNs for reconstitution after experimental destruction and to search for possible consequences of extracellular-matrix degradation for neurons and glial cells. The changes were induced by single intracortical injections of Proteus vulgaris chondroitinase ABC and studied after postinjection periods of 1 day to 5 months. The N-acetylgalactosamine-binding Wisteria floribunda agglutinin (WFA), an antibody against chondroitin-sulphate proteoglycans, three antibodies recognizing initial chondroitin or chondroitin sulphate moieties ('stubs') of proteoglycan core proteins, an antibody against the hyaluronan-binding protein component of versican, and biotinylated hyaluronectin, which binds to hyaluronan, were used as cytochemical markers. One day postinjection, the WFA-binding sites and hyaluronan were shown to be almost completely removed within a circumscribed digestion zone. The staining of different core-protein components revealed only fragments of PNs. These changes were found to be partly compensated 4 weeks after injection of chondroitinase ABC. After 8 and 12 weeks postinjection, the cytochemical and structural characteristics as well as the area-specific distribution patterns of PNs were progressively reconstituted. At 5 months postinjection, they could not be distinguished from those in untreated tissue. In contrast to such transient changes, a diffuse chondroitin-sulphate proteoglycan immunoreactivity persisted in the neuropil. Loss of neurons or alterations of their structure as well as reactions of glial cells were not observed. We conclude from this study that PNs, enzymatically destroyed in the adult rat brain, can be completely reconstituted, but the restoration of their extracellular-matrix components needs several months. PMID- 9746137 TI - Experimental jaw-muscle pain does not change heteronymous H-reflexes in the human temporalis muscle. AB - Muscle pain generally has an inhibitory effect on voluntary orofacial motor function. However, it is not known whether muscle pain causes direct or indirect changes in motoneuron excitability. In this study a monopolar needle stimulation technique was used to evoke the direct motor response (M-response) in the left masseter muscle and the heteronymous H-reflex in the left temporalis muscle as an indirect measure of motoneuron excitability. Series of 20 repeated electrical stimuli were delivered at 50% of maximal voluntary contraction (MVC) before, during, and after periods with experimental jaw-muscle pain in 11 healthy subjects. Pain was induced by standardized infusion of hypertonic (5%) saline into the mid-portion of the right masseter muscle. The mean pain intensity rating on a 100-mm visual analog scale was 42+/-5 mm. The short-latency responses (less than 6 ms) could be evoked in all subjects. Analysis of the latency and amplitude of the temporal H-reflex indicated no significant effect of jaw-muscle pain. The amplitude of the masseteric M-response was significantly smaller in the postpain condition than in the pain conditions (ANOVA, P=0.018), but no differences were found between the prepain and postpain conditions. In nine subjects, poststimulus periods (mean offset latency, 69.6+/-8.6 ms) with significantly (more than 50%) suppressed EMG activity were detected in the ipsilateral masseter muscle following the M-response (mean offset latency, 5.5+/-0.2 ms). These reflex responses did not show a systematic change during the pain conditions. In conclusion, acute contralateral jaw-muscle pain does not seem to modulate the motoneuron excitability as measured by the heteronymous H-reflex. PMID- 9746138 TI - Prolonged GABA(B) receptor-mediated synaptic inhibition in the cat spinal cord: an in vivo study. AB - In pentobarbitone-anaesthetised spinal cats, a comparison was made of the effects of intravenous bicuculline hydrochloride, a GABA(A)-receptor antagonist, and several (-)-baclofen (GABA(B)-receptor) antagonists (CGP 35348, 4638 , 56999A) on the prolonged inhibition of extensor-muscle monosynaptic reflexes, recorded from lumbar ventral roots, by brief or continuous tetanic stimulation of low-threshold afferent fibres of hindlimb flexor muscles. Two components of brief tetanus inhibition were detected. Whilst possibly of similar central latency, the inhibition associated with GABA(B) receptors had a longer time course than that reduced by bicuculline. Furthermore, whereas bicuculline reduced primary afferent depolarization, generated by the inhibitory volleys, and detected as dorsal-root potentials, such potentials were generally enhanced by intravenous baclofen antagonists. The inhibition of reflexes during and after continuous (333 Hz) tetanic flexor-nerve stimulation appeared to be predominantly associated with the activation of GABA(B) receptors. In the period following continuous tetanic flexor-nerve stimulation, during which monosynaptic extensor reflexes were reduced in amplitude, the action potentials of the intraspinal terminations of extensor-muscle group-Ia afferent fibres were reduced in duration, as detected by the time course of the recovery of the threshold to extracellular microstimulation following the arrival of an orthodromic impulse. A reduction in termination action-potential duration also accompanied the reduction by microelectrophoretic (-)-baclofen of the release of excitatory transmitter from group-Ia terminations, both presynaptic effects being blocked by microelectrophoretic baclofen antagonists. However, the reduction of the duration of the action potential of individual group-Ia terminations, which followed continuous flexor-nerve stimulation, was not sensitive to the baclofen antagonist CGP 55845A, but was diminished by bicuculline methochloride. Intravenously administered bicuculline hydrochloride, however, had little or no effect on the inhibition of reflexes following continuous flexor-nerve stimulation. These observations are discussed in the context of possible intraspinal pathways and pre- and postsynaptic mechanisms for GABA(A) and GABA(B) receptor-mediated inhibition of the monosynaptic excitation of spinal motoneurones and of the functional significance of central GABA(B) receptor-associated inhibitory processes, given the relatively minimal effects on motor activity and behaviour produced by baclofen antagonists that penetrate the mammalian blood-brain barrier. PMID- 9746139 TI - Neuroplasticity in the cat's visual system. Origin, termination, expansion, and increased coupling of the retino-geniculo-middle suprasylvian visual pathway following early ablations of areas 17 and 18. AB - We used anterograde and retrograde transsynaptic pathway tracing techniques to reveal the retinal origin and the cortical termination of the expanded retino geniculo-middle suprasylvian (MS) cortex pathway in adult cats which sustained lesions of areas 17 and 18 on the day of birth (P1) or at 1 month of age (P28). Following anterograde transsynaptic transport of tritiated amino acids from the eye, four major results were obtained: (1) a strong and specific pathway from retina through dorsal lateral geniculate nucleus (dLGN) to the posterior half of MS cortex was identified; this pathway is a substantial expansion of an insignificant pathway present in intact cats; (2) the terminus of the pathway was lower layer III and layer IV; (3) contralateral projections were stronger than ipsilateral projections; (4) projections in P28 cats were stronger than those in P1 cats. Following retrograde transsynaptic transport of WGA-HRP from posterior MS cortex, four additional results were obtained: (1) the pathway was enlarged and visuotopically organized; (2) the pathway arose primarily from alpha- and gamma-retinal ganglion cells; (3) a small number of beta-cells in P1 cats and a modest number in P28 cats also contribute to the pathway; (4) the combined numbers of gamma- and beta-cells relative to alpha-cells was greater in temporal retina than in nasal retina. The combined demonstration of both origin and terminus of the pathway with transsynaptic tracers argued strongly for high levels of coupling between primary and secondary pathway limbs in both P1 and P28 cats. This level of coupling, as well as other features of the pathway, have much in common with the retino-geniculo-17/18 pathway of intact cats. However, the retino-geniculo-MS system in P1 cats transmits primarily Y and W signals, in P28 cats X, Y, and W signals; whereas the retino-geniculo-17/18 pathway transmits primarily X and Y signals. These results have implications for understanding the repercussions of early visual cortex lesions in monkeys and humans. PMID- 9746140 TI - Learning of sequential movements by neural network model with dopamine-like reinforcement signal. AB - Dopamine neurons appear to code an error in the prediction of reward. They are activated by unpredicted rewards, are not influenced by predicted rewards, and are depressed when a predicted reward is omitted. After conditioning, they respond to reward-predicting stimuli in a similar manner. With these characteristics, the dopamine response strongly resembles the predictive reinforcement teaching signal of neural network models implementing the temporal difference learning algorithm. This study explored a neural network model that used a reward-prediction error signal strongly resembling dopamine responses for learning movement sequences. A different stimulus was presented in each step of the sequence and required a different movement reaction, and reward occurred at the end of the correctly performed sequence. The dopamine-like predictive reinforcement signal efficiently allowed the model to learn long sequences. By contrast, learning with an unconditional reinforcement signal required synaptic eligibility traces of longer and biologically less-plausible durations for obtaining satisfactory performance. Thus, dopamine-like neuronal signals constitute excellent teaching signals for learning sequential behavior. PMID- 9746141 TI - The effects of SDZ NKT 343, a potent NK1 receptor antagonist, on cutaneous responses of primate spinothalamic tract neurones sensitized by intradermal capsaicin injection. AB - Substance P, acting through neurokinin I receptors, is involved in the processing of nociceptive information in the spinal cord. Sensitization of spinothalamic tract neurons occurs to low-intensity stimuli following capsaicin injection. The current study tested the effects of the novel neurokinin I receptor antagonist, SDZ NKT 343, on the sensitization of spinothalamic tract cells by capsaicin in monkeys. Spinothalamic tract cells from the lumbar enlargement with receptive fields in the hindpaw were isolated and recorded before and after intradermal injection of capsaicin. The background activity and responses to brushing, pressing and pinching the skin were assessed. Thirty minutes after capsaicin injection there was an increase in background activity and responses to brush and pressure applied to the receptive field. Infusion of SDZ NKT 343 (for 30-45 min) significantly reversed the increased response to brushing without affecting the increased background activity or the increased response to pressure. Thus, blockade of neurokinin 1 receptors reduces the sensitized responses to innocuous mechanical stimuli but not to noxious mechanical stimuli. PMID- 9746142 TI - Analysis of direction-tuning curves of neurons in the turtle's accessory optic system. AB - Visual-movement sensitivity of neurons in the turtle's accessory optic system was investigated. Neuronal responses to stimulus direction and speed were analyzed to determine whether they reflect processing by a one-dimensional encoder of visual motion or whether they indicate directional integration of presynaptic direction sensitive responses whose maximal-response directions are distributed. Both of these mechanisms make predictions about the functional relationship between stimulus direction and response. The responses of single units in the basal optic nucleus to visual stimulation in different directions were described by both cosine and wrapped normal fitting functions. The wrapped normal function (a Gaussian curve mapped onto a circle) performed at least as well as the cosine function and described directional tuning curves of varying widths. Unlike cosines, the addition of two wrapped normals could describe multi-lobed directional data. Next, it was demonstrated that these neurons did not encode visual motion projected onto a single, spatial axis. Responses to the projected speed along the maximal-response direction were systematically lower than responses to the actual speed along that direction. Thus, for speeds above 1 degrees/s, neuronal response varies with respect to direction but not speed. Summation of presynaptic direction-sensitive responses with distributed maximal response directions (referred to as directional integration) is discussed as a means of accounting for these results. PMID- 9746143 TI - Facilitation of picture naming by focal transcranial magnetic stimulation of Wernicke's area. AB - On the basis of an evolutionary concept of language it was postulated that activation of the motor systems for arm movements, which are phylogenetically older, should facilitate language processes. In aphasic subjects picture naming can be improved by a concomitant movement of the dominant arm. In the present study it was investigated whether a similar facilitation can be observed in normal subjects by studying the effects of transcranial magnetic stimulation (TMS) on picture naming latencies. Suprathreshold focal TMS was applied to the left motor cortex for proximal arm muscles in right-handed subjects. The effects were compared with TMS of Wernicke's area. While TMS of the motor cortex and the non-dominant temporal lobe had no facilitatory effects, TMS of Wernicke's area decreased picture naming latencies significantly when TMS preceded picture presentation by 500 or 1000 ms. The observed effects depended on the intensity of the stimulus used. While clearly present with intensities of 35% and 55% of maximum output the facilitation disappeared with higher stimulation intensities. It is concluded that focal magnetic stimulation is able to facilitate lexical processes due to a general preactivation of language-related neuronal networks when delivered over Wernicke's area. PMID- 9746144 TI - Anodally focused polarization of peripheral nerve allows discrimination of myelinated and unmyelinated fiber input to brainstem nuclei. AB - We investigated the ability of a novel direct current (DC) polarization technique to block selectively the conduction in peripheral myelinated nerve fibers and allowing propagation in only unmyelinated fibers. In anesthetized adult rats, distal branches of the sciatic nerve (caudal cutaneous sural and tibial nerves) were exposed for electrical stimulation of A- and C-fibers. Two specially fabricated trough electrodes of different size and surface area were placed onto the sciatic nerve. Through these proximal electrodes a controlled ramped DC was timed to coincide with the arrival of A- and C-fiber action potentials, evoked electrically at the distal nerves or naturally from the foot or ankle, with the intent of blocking propagation in A-fibers while allowing C-fiber throughput. Neuronal recordings were made both peripherally (proximal sciatic nerve fascicles or L5 dorsal roots) and centrally (single cells in the nucleus gracilis or nucleus reticularis gigantocellularis). The DC polarization was shown to block conduction in myelinated A-fibers effectively, while allowing conduction in the unmyelinated C-fibers, without activation of fibers via the DC polarization itself. This was dependent upon the following factors: electrode polarity, onset rate of polarization, peak amplitude of polarization, distance between polarizing electrodes, size difference between polarizing electrodes, and gross nerve size. These experiments demonstrate that anodally focused DC polarization, applied utilizing two trough electrodes of different sizes, is capable of effectively, reversibly, and reproducibly blocking conduction in myelinated A-fibers evoked either electrically or naturally, while still allowing conduction to occur in the unmyelinated C-fiber population. In the context of experimental usage, we have demonstrated blocking of low-threshold A-fiber, but not C-fiber, mediated inputs to the caudal brainstem. This technique should find wide application in studies involving the processing of information conveyed centrally by the unmyelinated C fiber afferent population, including discriminating afferent responses to peripheral stimuli, the role of C-fiber input in reflex activity, and the plasticity following injury or other manipulations. PMID- 9746145 TI - Age-related performance of human subjects on saccadic eye movement tasks. AB - We measured saccadic eye movements in 168 normal human subjects, ranging in age from 5 to 79 years, to determine age-related changes in saccadic task performance. Subjects were instructed to look either toward (pro-saccade task) or away from (anti-saccade task) an eccentric target under different conditions of fixation. We quantified the percentage of direction errors, the time to onset of the eye movement (saccadic reaction time: SRT), and the metrics and dynamics of the movement itself (amplitude, peak velocity, duration) for subjects in different age groups. Young children (5-8 years of age) had slow SRTs, great intra-subject variance in SRT, and the most direction errors in the anti-saccade task. Young adults (20-30 years of age) typically had the fastest SRTs and lowest intra-subject variance in SRT. Elderly subjects (60-79 years of age) had slower SRTs and longer duration saccades than other subject groups. These results demonstrate very strong age-related effects in subject performance, which may reflect different stages of normal development and degeneration in the nervous system. We attribute the dramatic improvement in performance in the anti-saccade task that occurs between the ages of 5-15 years to delayed maturation of the frontal lobes. PMID- 9746146 TI - The role of the pedunculopontine tegmental nucleus in relation to conditioned motor performance in the cat. I. Context-dependent and reinforcement-related single unit activity. AB - The activity of the pedunculopontine tegmental nucleus (PPTg) neurons was recorded in three unrestrained cats operantly conditioned to perform a lever release movement. The movement had to be initiated either rapidly after a (click) stimulus in a simple reaction-time paradigm or had to be delayed after the same stimulus in trials identified by a tone cue. Successful trials were rewarded by a food pellet. A total of 107 neurons were recorded with microelectrodes. Brief spike neurons (mean duration: 0.7 ms) and broad spike neurons (mean duration: 2 ms) presumed to be cholinergic were detected. Of the 73 neurons localized in the PPTg area, 53 had brief spikes and 20 broad spikes. Changes in activity most commonly occurred very early after the stimulus or during the reinforcement process. Most neurons with brief spikes exhibited very early excitation after the stimulus and reinforcement-related activity. These neurons had a mean activity of 23.7 impulses/s in the period preceding the stimulus. The onset of activation after the stimulus had a latency of 8.6+/-6.9 ms (mean+/-SD), with a range of 4 35 ms. In trials where the movement had to be delayed after the stimulus, the early activation disappeared or was considerably reduced, showing that it was context-dependent. A small proportion of neurons with brief spikes initially decreased activity after the stimulus, but with a latency >9 ms. All the neurons with broad spikes, except one, had reinforcement-related activity. Half of them showed exclusively reinforcement-related activity, the other half also early activation after the stimulus. These neurons were about half as active in the period preceding the stimulus occurrence than the neurons with brief spikes. The early context-dependent activation is discussed in relation to the excitatory projection of PPTg neurons on the subthalamic nucleus. The reinforcement-related activity, preferentially evidenced in broad spike neurons presumed to be cholinergic, is speculated to be associated with cholinergic projection of PPTg neurons to the dopaminergic neurons of the substantia nigra. Finally, the role of PPTg in the ongoing control of motor performance and reinforcement processes is discussed in relation to the basal ganglia circuitry. PMID- 9746147 TI - The role of the pedunculopontine tegmental nucleus in relation to conditioned motor performance in the cat. II. Effects of reversible inactivation by intracerebral microinjections. AB - The effects of reversible pharmacological manipulation of the neuronal activity in the pedunculopontine tegmental nucleus (PPTg) on the performance of a conditioned movement was studied in two freely moving cats. The microinjections were given in regions where, in the same subjects, we had previously identified neurons with context-dependent early activity after a trigger stimulus and with reinforcement-related activity. The subjects were conditioned to perform a forelimb-flexion movement controlled by a simple reaction-time task. In addition, one subject was trained to execute the same flexion movement, but delayed after the trigger stimulus. Food pellets were used as the reinforcer. Lidocaine injections (1 microl of 2% solution, injected over a 6-min period) induced a transient arrest of performance within minutes. The cessation of performance could be preceded by behavioral signs such as meowing, attempt to escape from the experimental booth, licking, or stereotyped posture. No rotational behavior could be observed. The effects of lidocaine could be mimicked in one subject by an extinction procedure. Muscimol injections (two injections of 0.2 microg in 1 microl, tested in one subject) also induced arrest of performance, but the return to pre-injection level of performance could not be obtained within the time of the test session. The quantitative analysis of reaction times and of inter-trial intervals showed that altering PPTg activity affected inter-trial intervals, but only slightly affected the reaction times. It is speculated that the PPTg is involved in the reinforcement process related to selecting the appropriate motor program. PMID- 9746148 TI - Reaction of Muller cells after increased intraocular pressure in the rat retina. AB - Using light microscopy and immunocytochemistry, we investigated the morphological changes of retinal tissues and the reaction of Muller cells in the ischemic rat retina induced by increasing intraocular pressure. At early stages (from 1 h to 24 h after reperfusion), cells in the ganglion cell layer and in the inner nuclear layer showed some degenerative changes, but at later stages (from 72 h to 4 weeks) marked degenerative changes occurred in the outer nuclear layer (ONL). At 4 weeks after reperfusion, the ONL was reduced to 1 or 2 cell layers. Immunoreactivity for glial fibrillary acidic protein (GFAP) appeared in the endfeet and distal processes of Muller cells as of 1 h after reperfusion. GFAP immunoreactivity in Muller cells increased up to 2 weeks and then decreased at 4 weeks after reperfusion. Our findings suggest that Muller cells are involved in the pathophysiology of retinal ischemia through the expression of GFAP. The degree of GFAP expression in Muller cells closely correlated with that of the degeneration of retinal neurons. PMID- 9746149 TI - Sound-evoked efflux of excitatory amino acids in the guinea-pig cochlea in vitro. AB - We have used the perfused guinea-pig temporal-bone preparation to study the sound evoked efflux of aspartate and glutamate, which are putative afferent transmitters in the cochlea. The cochlea was stimulated with white noise at 89, 95, and 101 dB SPL. Cochlear function was monitored by recording the endocochlear potential, the cochlear microphonic, and the summating potential. In silence, there was a low basal efflux of both amino acids. A significant and intensity dependent sound-evoked efflux of aspartate was observed at all levels, whereas a significant efflux of glutamate was found only at the 101 dB SPL level. Immunohistochemistry of sections from the organ of corti showed an ubiquitous distribution of glutamate-like immunoreactivity in the sensory organ and ganglion, whereas aspartate-like immunoreactivity was found in the region of the inner hair cells and in the spiral ganglion. In view of these findings, we suggest that not only glutamate, but also aspartate may have a neurotransmitter role in the afferent pathway of the cochlea. PMID- 9746150 TI - Retrograde tracing studies of subdivisions of the orbicularis oculi muscle in the rhesus monkey. AB - Functionally and anatomically, the orbicularis oculi (OO) muscle can be subdivided in a pretarsal, a preseptal, and an orbital portion. In the rhesus monkey, fluorescent and neuronal retrograde tracing experiments were performed in the pretarsal or the orbital portion of the OO muscle, or both, using fast blue, diamidino yellow, and wheat germ agglutinin-horseradish peroxidase as tracers. The preseptal portion was not investigated because of close anatomical relationships to the other portions. It was found that motoneurons innervating the OO muscle are located exclusively within the intermediate subnucleus of the motor facial nucleus. The upper pretarsal motoneurons show a specific distribution in the dorso-rostral border area of the intermediate subnucleus, representing a dome-like organization, while lower pretarsal motoneurons are situated more ventrally in the adjacent area. The pretarsal motoneurons are all located dorsally in the rostral half and the upper part of the caudal half of the intermediate subnucleus. The upper pretarsal portion is subserved by about one third of the total intermediate motoneuron population. The size of the upper pretarsal motoneurons is similar to that of the motoneurons of the lower pretarsal portion of the OO muscle and falls, for the vast majority, into the large motoneuronal range. Motoneurons belonging to the upper and lower orbital portions are located ventrally and are more randomly distributed in the rostral half of the intermediate subnucleus. The size of orbital motoneurons varies from small to large. The large fraction of pretarsal motoneurons may reflect the specific function of the upper pretarsal portion during rapid and highly coordinated movements of the eyelids in different types of blinking. PMID- 9746151 TI - Presaccadic omnidirectional burst activity in the basal interstitial nucleus in the monkey cerebellum. AB - We recorded saccade-related neurons in the vicinity of the dentate nucleus of the cerebellum in two monkeys trained to perform visually guided saccades and memory guided saccades. Among 76 saccade-related neurons, 38 showed presaccadic bursts in all directions. More than 80% of such burst neurons were located in the area ventral to, not inside, the dentate nucleus, which corresponded to the basal interstitial nucleus (BIN as previously described). We found that the activity of the BIN neurons was correlated with saccade duration but not with saccade amplitude or velocity. Thus, when tested with visually guided saccades, the burst started about 16 ms before saccade onset and ended about 33 ms before saccade offset, regardless of saccade amplitude. The characteristic timing of the BIN cell activity was maintained for different types of saccades (visually guided, memory-guided and spontaneous saccades), which had different dynamics. Although the number of spikes in a burst for each neuron was linearly correlated with saccade amplitude for a given type of saccade, the slope varied depending on the type of saccade. Peak burst frequency was uncorrelated with saccadic peak velocity. In contrast, burst duration was highly correlated with saccade duration regardless of the type of saccade. These results suggest that BIN neurons may carry information to determine the timing of saccades. PMID- 9746152 TI - Effects of mesaconitine on [3H]noradrenaline uptake and neuronal excitability in rat hippocampus. AB - Mesaconitine, one of the main alkaloids contained in Aconiti tubers, is a centrally acting analgesic without affinity to opioid receptors. It has been reported that the antinociception is due to an interaction with the noradrenergic system. In the present study, the effect of mesaconitine on the uptake of noradrenaline and on neuronal activity was examined in rat hippocampus. Experiments were performed as a study of [3H]noradrenaline uptake into rat hippocampal synaptosomes. Mesoconitine inhibited [3H]noradrenaline uptake in a concentration-dependent manner with a Ki of 111.95+/-18 nM. In a further series of experiments, the effects of mesaconitine on the extracellularly recorded population spike were investigated in rat hippocampal slices. At a concentration of 10 nM, mesaconitine increased the amplitude of the postsynaptic population spike by 31.10%+/-6.7% of control and elicited one or two additional spikes. The presynaptic fiber spike and the field excitatory postsynaptic potential were not affected by this alkaloid. The enhancement of neuronal activity was abolished by 1 microM propranolol as well as by 1 microM timolol. It is concluded that mesoconitine increased the excitability in rat hippocampal pyramidal cells by an involvement of the noradrenergic system, with at least one mechanism being inhibition of noradrenaline uptake leading to an enhanced extraneuronal noradrenaline level. PMID- 9746153 TI - Unilateral testing of utricular function. AB - A modified rotatory chair test is reported in which radial acceleration, generated by eccentric displacement of the subject during constant angular velocity, is exploited as a unilateral stimulation to the otolith organs. During constant angular rate rotation, the test subject is displaced laterally on the rotating turntable by 3.5 cm, so that one labyrinth becomes aligned with the rotatory axis while the second - eccentric - labyrinth is solely exposed to the altered gravito-inertial acceleration (GIA). Previously reported results showed that the direction of the response is independent of the direction of turntable rotation, ruling out any canal influence, and indicated that in a normal population the response, measured in one eye, was symmetrical for displacement of the left and right labyrinths. This mode of stimulus thus appears to elicit a unilateral otolith-ocular response (OOR). Examination of this unilateral OOR was extended in the present study; comparative testing with head-tilt to gravity, i.e. involving bilateral stimulation to the otolith organs, was carried out. Movements of both eyes were recorded (by three-dimensional video-oculography), in order to examine response conjugacy. To verify the specificity of the unilateral stimulus, tests were performed with patients who had previously undergone unilateral section of the vestibular nerve as treatment for acoustic neuroma. The eccentric displacement profile (EDP) and head-tilt stimulus each included ten cycles of left-right oscillation in order to permit signal averaging. In the normal subjects (n=12) the torsional component of the OOR proved to be both labyrinth-symmetrical and conjugate, during both bilateral and unilateral otolith stimulation. OOR gain (ocular torsion/GIA tilt) was higher for bilateral than unilateral stimulation. Bilateral OORs, obtained from three of the five unilaterally deafferented patients, proved less symmetrical and conjugate than in the normals. Unilateral OORs in all five patients were characteristically asymmetrical, with little or no response during stimulation of the diseased labyrinth. PMID- 9746154 TI - The role of learned pictorial cues in the programming and control of grasping. AB - Binocular information has been shown to be important for the programming and control of reaching and grasping. Even without binocular vision, people are still able to reach out and pick up objects accurately - albeit less efficiently. It remains unclear, which of the many available monocular depth cues humans use to calibrate manual prehension when binocular information is not available. In the present experiment, we examined whether or not subjects could use a learned relationship between the elevation of a goal object in the visual scene and its distance to help program and control the required grasp. The elevation of the goal object was systematically varied with distance in some blocks of trials by presenting the object at different positions along a horizontal plane 35 cm below eye level. In other blocks of trials, elevation did not vary with distance because the objects were always presented along the subject's line of sight. When subjects viewed these two displays monocularly, they showed fewer on-line adjustments in the trajectory of the limb and the aperture of the fingers when the elevation of the target object in the visual scene could be used to help program the required movements. No such difference between performance on the two arrays was seen when subjects were allowed a full binocular view. This study confirms that subjects are indeed able to use a learned relationship between the elevation of an object and its distance as a cue for programming grasping movements when binocular information is not available. Together with evidence from work with neurological patients who have difficulty perceiving pictorial cues, these findings suggest that the visuomotor system might normally "prefer" to use binocular cues, but can fall back on learned pictorial information when binocular vision is denied. PMID- 9746155 TI - Spatial-frequency-related efficacy of visual stabilisation of posture. AB - The present study investigates the efficacy of visual stabilisation of posture for different spatial frequencies of a visual stimulus. Circular sine wave gratings were used to analyse the correlation between perception of motion in depth and stabilisation of fore-aft sway by the mechanism of detecting changes in target size. Body sway was recorded by a force-measuring platform (series A) and, in addition, by simultaneous tracking of infrared markers fixed to the subject's body (series B). Mean velocity and amplitude (RMS) of body sway were calculated in both sagittal (a-p) and lateral (l-r) planes. Sagittal sway was of least magnitude when viewing contrast gratings with lowest thresholds, whereas higher thresholds resulted in increasing sway parameters. As intended by the design of the stimuli, sagittal sway was correlated closer with the stabilising effect exerted by the different stimuli than was lateral sway. Sway velocity was reduced more efficiently, however, with a lower correlation with the psychophysical transfer function, than was RMS sway. Since sway velocity measured by the platform is suggested to depend to a greater extent on dynamic muscle forces generated at each individual body site the results indicate that visual information can be used to reduce and thereby optimise dynamic muscle action (sway velocity) even though static body sway is either not or less reduced. A comparable economisation of sway velocity but not of RMS sway was also seen at the end of posture investigations, indicative of positive training effects. PMID- 9746156 TI - Interactions between vestibular and proprioceptive inputs triggering and modulating human balance-correcting responses differ across muscles. AB - Interactions between proprioceptive and vestibular inputs contributing to the generation of balance corrections may vary across muscles depending on the availability of sensory information at centres initiating and modulating muscle synergies, and the efficacy with which the muscle action can prevent a fall. Information which is not available from one sensory system may be obtained by switching to another. Alternatively, interactions between sensory systems and the muscle to which this interaction is targeted may be fixed during neural development and not switchable. To investigate these different concepts, balance corrections with three different sets of proprioceptive trigger signals were examined under eyes-open and eyes-closed conditions in the muscles of normal subjects and compared with those of subjects with bilateral peripheral vestibular loss. The different sets of early proprioceptive inputs were obtained by employing three combinations of support surface rotation and translation, for which ankle inputs were nulled, normal or enhanced, the knees were either locked or in flexion, and the trunk was either in flexion or extension. Three types of proprioceptive and vestibulospinal interactions were identified in muscles responses. These interactions were typified by the responses of triceps surae, quadriceps, and paraspinal muscles. The amplitudes of stretch responses at 50 ms after the onset of ankle flexion in triceps surae muscles were related to the velocity of ankle stretch. The amplitude of balance-correcting responses at 100 ms corresponded more with stretch of the biarticular gastrocnemius when the knee was re-extended at 60 ms. Absent stretch reflexes at 50 ms in triceps surae with nulled ankle inputs caused a minor, 12-ms delay in the onset of balance correcting responses in triceps surae muscles. Vestibular loss caused no change in the amplitude of balance-correcting responses, but a negligible decrease in onset latency in triceps surae even with nulled ankle inputs. Stretch responses in quadriceps at 80 ms increased with the velocity of knee flexion but were overall lower in amplitude in vestibular loss subjects. Balance-correcting responses in quadriceps had amplitudes which were related to the directions of initial trunk movements, were still present when knee inputs were negligible and were also altered after vestibular loss. Stretch and unloading responses in paraspinals at 80 ms were consistent with the direction of initial trunk flexion and extension. Subsequent balance-correcting responses in paraspinals were delayed 20 ms in onset and altered in amplitude by vestibular loss. The changes in the amplitudes of ankle (tibialis anterior), knee (quadriceps) and trunk (paraspinal) muscle responses with vestibular loss affected the amplitudes and timing of trunk angular velocities, requiring increased stabilizing tibialis anterior, paraspinal and trapezius responses post 240 ms as these subjects attempted to remain upright. The results suggest that trunk inputs provide an ideal candidate for triggering balance corrections as these would still be present when vestibular, ankle and knee inputs are absent. The disparity between the amplitudes of stretch reflex and automatic balance-correcting responses in triceps surae and the insignificant alteration in the timing of balance correcting responses in these muscles with nulled ankle inputs indicates that ankle inputs do not trigger balance corrections. Furthermore, modulation of balance corrections normally performed by vestibular inputs in some but not all muscles is not achieved by switching to another sensory system on vestibular loss. We postulate that a confluence of trunk and upper-leg proprioceptive input establishes the basic timing of automatic, triggered balance corrections which is then preferentially weighted by vestibular modulation in muscles that prevent falling. (ABSTRACT TRUNCATED) PMID- 9746158 TI - The evolution of stigmata of hemorrhage in bleeding peptic ulcers: a sequential endoscopic study. AB - BACKGROUND AND STUDY AIMS: Stigmata of hemorrhage in bleeding peptic ulcers have prognostic characteristics. In the present study, the evolution of these stigmata was studied prospectively using daily endoscopic examinations. PATIENTS AND METHODS: From January 1989 to October 1989, 778 consecutive patients with bleeding peptic ulcers underwent endoscopy within 24 hours of admission. The bleeding peptic ulcers were assigned by three endoscopists to five categories, those with: a) active bleeding, b) a nonbleeding visible vessel, c) adherent clot, d) dot, or e) a clean base. Actively bleeding ulcers were treated by epinephrine injection. Ulcers with nonbleeding visible vessels, adherent clots, or dots were left untreated. Daily endoscopic examinations were carried out for three subsequent days, or until the ulcer base became clean. RESULTS: On day 0, there were 56 actively bleeding ulcers (7%), 62 ulcers with visible vessels (8%), 104 with adherent clots (13%), 182 with flat dots (23%), and 374 with a white base (48%). On the subsequent three days, 24 of 62 ulcers with visible vessels (39%), 30 of 104 with adherent clots (29%), 24 of 182 with dots (13%), and 19 of 374 with a clean base (5%) on day 0 re-bled endoscopically or clinically, or both. The overall rebleeding risk was 9.9%, 4.9%, and 2.7% on days 1, 2, and 3, respectively. CONCLUSIONS: Stigmata of hemorrhage in bleeding peptic ulcers are predictive of rebleeding. They represent intermediate phases in the evolution of bleeding vessels into clean-based ulcers. The associated rebleeding risk diminishes as the vessel disappears from the ulcer base. PMID- 9746157 TI - Non-variceal upper gastrointestinal bleeding and Forrest's classification: diagnostic agreement between endoscopists from the same area. AB - BACKGROUND AND STUDY AIMS: The lack of uniformity in defining the stigmata of hemorrhage in patients with bleeding ulcers is suggested by the wide range among published studies in prevalence and rebleeding rates for the same stigmata. Moreover there is, in published trials of endoscopic hemostasis, little standardization of definitions of stigmata of hemorrhage. The aim of this study was to assess the interobserver agreement among endoscopists from the same area (Piedmont and Valley of Aosta). PATIENTS AND METHODS: A workshop for 47 expert endoscopists was organized in order to evaluate their agreement in the diagnosis of stigmata of recent hemorrhage, according to Forrest's classification. During the meeting 25 videotapes from endoscopic examinations of patients with recent non-variceal bleeding were shown to the 47 endoscopists, who were asked to classify every lesion. RESULTS: The overall and beyond chance interobserver agreement was calculated by means of the kappa statistic. The overall agreement among endoscopists was highly significant (p < 0.001, kappa=0.60), while the beyond chance agreement varied from excellent to good for lesions with active bleeding (kappa=0.76 and kappa=0.61 for FIA and FIB lesions respectively), whereas for lesions with stigmata of recent hemorrhage kappa varied from 0.44 to 0.49. CONCLUSIONS: These data suggest the need for better knowledge of endoscopic criteria, in order to evaluate the results of endoscopic therapy and to assess new treatments. PMID- 9746159 TI - Fibrin sealing in peptic ulcer bleeding: the fate of the clot. AB - BACKGROUND AND STUDY AIMS: The injection of fibrin tissue glue is a promising endoscopic method for hemostasis of peptic ulcer bleeding. So far, no clinical study has focused on the ulcer healing process after endoscopic fibrin injection. PATIENTS AND METHODS: A morphological study was performed on all resection specimens from patients operated on between 1 January 1994 and 31 December 1996 for gastroduodenal ulcer bleeding with prior endoscopic injection of fibrin glue. The fibrin clot was characterized histologically for its size, location and aspect. RESULTS: Of 227 patients endoscopically treated with a double-lumen needle, 20 underwent resection. The interval between fibrin injection and resection ranged from 6 hours to 9 days. In 15 patients the ulcer was identified in the resection specimen. No fibrin remnants were detectable in three, sparse fibrin deposits were seen in eight and large amounts of fibrin were noted in the submucosa or subserosa in four specimens. With increasing time, the clot was gradually organized by phagocytes and angio-fibroblasts and was finally replaced by endogenous granulation tissue. An exuberant or tissue-destructive reaction did not appear. CONCLUSIONS: In this study, endoscopic fibrin sealing of bleeding ulcers resulted in appearance of a bland fibroblast-rich granulation tissue. The depth of fibrin glue injection is difficult to standardize with the aid of double lumen needles. PMID- 9746160 TI - Improved endoscopic stenting for malignant dysphagia using Tygon plastic prostheses. AB - BACKGROUND AND STUDY AIMS: Endoscopic palliative treatment of malignant esophageal stenosis using conventional plastic stents has been reported to be associated with a considerable risk of perforation. Stenoses with a distance of less than 2cm from the upper esophageal sphincter (UES) have generally been excluded from treatment. Using self-expandable metal stents, procedure-related complications are rare. However, the rates of late complications necessitating retreatment appear to be as high as those of plastic stents. This study describes our stent placement technique and our results using a modified Tygon plastic stent. PATIENTS AND METHODS: Over a two-year period, 71 consecutive patients with incurable malignant esophageal stenosis were prospectively studied. Tygon plastic stents of diameter 9-14 mm were individually tailored according to length and location of the stenosis. Prior to stenting, stepwise bougienage was performed, if necessary over several sessions. After endoscopic placement of a guide wire, the stent was inserted over a bougie without fluoroscopic monitoring. RESULTS: A total of 71 patients (54 men and 17 women, median age 69, range 34-93), were treated with Tygon plastic stents (14 mm: 19 patients; 12 mm: 50 patients; 9 mm: 2 patients). Median length of the strictures and of the stents were 7 (range 2 18) and 10 (range 6-25) cm, respectively. Four patients had an associated esophago-respiratory fistula. After a median of 2 (range 1-5) bougienage sessions, stent insertion was technically successful in all patients. Forty-one stents were placed across the cardia, 13 were positioned 0.5-1 cm below the UES. Three patients had to undergo retreatment within 24 hours because of pain or stent migration and the stents were repositioned or exchanged. No procedure related perforation, hemorrhage or respiratory problems were observed. During a median follow-up of 63 (range 2-388) days, 82% of the patients died. Improvement or stabilization of dysphagia allowing for oral nutrition could be achieved in 89%. Dislocation occurred in eight patients, bolus obstruction in five patients and tumor overgrowth in four patients. Three of the four fistulas could be covered by the stent. In one patient with a fistula located at the level of the UES, a stent was placed but migrated after 5 days. Overall, 27 patients (38%) required reinterventions, mainly for dysphagia or nutritional problems. CONCLUSIONS: In our experience, Tygon plastic stents with a diameter of 9-14 mm can be safely placed after stepwise, less extensive bougienage. Effective palliation is possible even for lesions located close to the UES. Perforation can be avoided. Reintervention rates seem to be comparable to those seen with self expanding metal stents. PMID- 9746161 TI - Endoscopic placement of indigenous plastic esophageal endoprostheses--does it still have a role in the era of expandable metallic stents? A prospective Indian study in 265 consecutive patients. AB - BACKGROUND AND STUDY AIMS: Esophageal endoprosthesis placement is an established method of palliating inoperable esophageal malignancy. However, the prosthesis choice varies, with expandable metal stents recently gaining popularity. We present our experience of using an indigenously developed plastic prosthesis in 265 patients prospectively in the period April 1992 to May 1996. PATIENTS AND METHODS: An indigenous endoprosthesis made of a medical grade, nontoxic, radiopaque plastic material was placed successfully in 259 patients after serial dilatation of the malignant stricture. Patients were followed up once every month for at least 6 months and also in between if they developed any significant symptoms. The results were analyzed prospectively with special emphasis on the cost of the therapy, technical success of placement, improvement of swallowing and occurrence of complications. RESULTS: The technical success of placement was 97.7% (259/265 patients). The mean dysphagia score improved from 3.2 to 1.2; 212 patients (81.8%) could swallow semisolids whereas 47 patients (18.2%) could swallow liquids. Though 75 patients (28.3%) had an associated tracheoesophageal fistula and 29.8% had received prior radiotherapy/chemotherapy, immediate complications like perforation, respiratory distress or severe hemorrhage were encountered in only 4.3% of patients. Late complications occurred in 12.7% and 32.8% of the patients complained of mild post-procedure pain in the chest. The overall procedure-related mortality was 3.9%. The average cost of the prosthesis was only US$ 15 per patient. CONCLUSIONS: Placement of a plastic prosthesis is still a very effective and safe method for relief of malignancy-induced dysphagia. The associated complications can be significantly reduced by modifying the prosthesis material/design and adhering to a careful technique. The extremely low cost of the prosthesis and its safety profile makes this treatment highly cost-effective and widely applicable in developing countries such as India. PMID- 9746162 TI - The contribution of endoscopy and biopsy to the diagnosis of periampullary tumors. AB - BACKGROUND AND STUDY AIMS: Endoscopy and biopsy from a suspicious Vater's papilla may establish an early preoperative diagnosis of a periampullary tumor. However, information regarding the diagnostic accuracy of this procedure is limited and variable. The aim of the present study was to evaluate retrospectively the accuracy of this procedure compared to that of other diagnostic methods. PATIENTS AND METHODS: Among 928 patients referred to our institute for endoscopic retrograde cholangiopancreatography (ERCP), a suspicious Vater's papilla was seen in 28. In each case comparison was made between the pre-ERCP clinical diagnosis, endoscopic appearance, histologic interpretation of endoscopic biopsies, and the final diagnosis. Two patients in whom a final diagnosis was not available were excluded from the study. RESULTS: A final diagnosis of an ampullary or periampullary carcinoma was established in 17 patients (65%), a carcinoma within an adenoma of the papilla in three patients (12%), and adenoma and a metastatic gallbladder carcinoma in one patient each. The remaining four patients (15%) were finally diagnosed as having "pseudotumors" (due to choledocholithiasis). Eight (38%) of the 21 patients with ampullary or periampullary neoplasm also had gallstones. A pre-ERCP diagnosis (by clinical evaluation and non-invasive imaging) of tumor versus choledocholithiasis was accurate in only 65% of all 26 patients. In these, the diagnostic accuracy of endoscopic appearance and endoscopic biopsy was 77% and 85%, respectively. Regarding the 21 patients with carcinomas, the diagnosis by endoscopic appearance was more accurate than that by endoscopic biopsy (90% vs 81%). Unlike the positive predictive values, the negative predictive values for malignancy were weak: 33% for the endoscopic appearance and 50% for the endoscopic biopsy. CONCLUSIONS: Because of a high incidence of concurrent cholelithiasis, many patients with a periampullary tumor seen during ERCP are misdiagnosed earlier (by clinical evaluation and non invasive imaging) as having choledocholithiasis only. However, the accuracy of endoscopy and biopsy is also limited. This limitation must be considered when evaluating the optimal management of patients with suspected periampullary tumor. PMID- 9746163 TI - Endoscopic diathermy in patients with cardiac pacemakers. AB - BACKGROUND AND STUDY AIMS: Malfunction of cardiac pacemakers related to diathermy in surgical procedures has been reported, but the risks of endoscopic diathermy in pacemaker patients is unknown. The aim of this study was to investigate current practice amongst British gastroenterologists regarding endoscopy in cardiac pacemaker patients. METHODS: An anonymous postal questionnaire survey of 634 members of the Endoscopy section of the British Society of Gastroenterology was conducted. RESULTS: 410/634 (65%) replied. Respondents conducted 59270 endoscopic retrograde cholangiopancreatography procedures (ERCPs) and 88544 colonoscopies per year. 77.3% of respondents were aware of the possibility of adverse interactions between diathermy and pacemakers. 74.2% enquired whether a pacemaker was present prior to endoscopy. In cases where patients were known to have pacemakers fitted, 23.9% recorded an electrocardiogram (ECG) prior to endoscopic diathermy, 36.2% conducted ECG monitoring during the procedure, 35.9% consulted a cardiologist or pacemaker technician and 13.4% carried out specific preventative measures. 4.1% of all respondents were aware of instances of pacemaker malfunction having occurred during endoscopic diathermy. CONCLUSION: Most gastroenterologists surveyed were aware of the possibility of adverse interactions between diathermy and cardiac pacemakers, but few undertook measures to detect or prevent pacemaker malfunction. Endoscopic diathermy in cardiac pacemaker patients, however, appears generally safe, although the endoscopist should be aware of the small chance of an adverse interaction. PMID- 9746164 TI - An assessment of local curability of endoscopic surgery in early gastric cancer without satisfaction of current therapeutic indications. AB - BACKGROUND AND STUDY AIMS: Therapeutic endoscopy for early gastric cancer has been established with strict criteria for indications. In the present study, we evaluated the efficacy of endoscopic treatment in cases that did not fulfil the standard therapeutic criteria, consisting of well differentiating mucosal adenocarcinomas less than 2 cm in size and without an ulcer or a scar. PATIENTS AND METHODS: Sixty nine early gastric cancers in 64 patients that did not fulfil the standard criteria were treated endoscopically, and a rate of cure was retrospectively assessed during a mean follow-up of 5.2 years. Endoscopic treatment consisted of mucosal resection or thermal methods, or both. RESULTS: Curative resection was achieved in 19/20 (95%) of cases which came into one of the following categories, all being well differentiated adenocarcinomas, less than 3.0 cm in size, without ulcer or the scar of an ulcer, with invasion limited to mucosal layer (depth m); tumors less than 2.0 cm, with an ulcer or scar, depth m; tumors less than 2.0 cm, without ulcer or scar, invading the submucosa but in which invasion was limited to the superficial portion (depth sm-1); and poorly differentiated tumors less than 1.0 cm in size, without an ulcer or scar, depth m. The rate of cure in this group was statistically similar to the cure rate of cases that fulfilled the standard criteria (98%). CONCLUSIONS: Our retrospective results suggest that the indications for curative treatment of early gastric cancer could be expanded. Prospective studies are required. PMID- 9746165 TI - Self-expandable metal stents for malignant gastric outlet obstruction: a modified technique. AB - BACKGROUND AND STUDY AIMS: Endoscopic palliative treatment may be effective in the management of malignant gastric outlet obstruction. However, experience in this area is limited, and the techniques vary widely. In this retrospective study, a uniform technique using nearly identical self-expandable metal stents was employed to assess technical feasibility, safety, and outcome. PATIENTS AND METHODS: Eight patients presenting with clinical findings of gastric outlet obstruction confirmed by upper gastrointestinal radiography underwent endoscopic placement of expandable metal stents. All patients had primary or metastatic malignancy involving the pylorus or duodenum. Endoscopic and Gastrografin enhanced upper gastrointestinal radiographic evaluations were carried out immediately after stent placement. Complications and clinical outcomes were assessed in each patient. RESULTS: Five patients had extrinsic compression of the descending duodenum due to pancreatic cancer, two had pyloric stenosis from metastatic cancer, and one patient had primary duodenal cancer. Stent placement was successful in all patients, and was followed by clinical improvement. There was one death within 30 days, related to pneumonia. CONCLUSION: Endoscopic self expandable stent placement appears to be a reasonable therapeutic alternative in patients with malignant gastric outlet obstruction. PMID- 9746166 TI - The role of therapeutic endoscopy associated with extracorporeal shock-wave lithotripsy and bile acid treatment in the management of Caroli's disease. AB - BACKGROUND AND STUDY AIMS: Caroli's disease causes relapsing episodes of cholangitis due to the presence of intrahepatic lithiasis. Strategies for cholangitis prevention are still widely debated. Ursodeoxycholic acid, hepaticojejunostomy, partial hepatectomy, or transplantation, have all been proposed as therapeutic options. The aim of this study was to evaluate the role of therapeutic endoscopy, and especially endoscopic sphincterotomy (ES), in the management of Caroli's disease. PATIENTS AND METHODS: Between 1983 and 1995, six patients with Caroli's disease (mean age 52, range 17-75) underwent endoscopic retrograde cholangiopancreatography (ERCP) for acute cholangitis. Sphincterotomy was performed if common bile duct stones were present. Extracorporeal shock-wave lithotripsy, (ESWL) or intraductal electrohydraulic lithotripsy (IEL) were performed if necessary. RESULTS: The mean number of endoscopic sessions per patient was four (range three to seven). Sphincterotomy was performed in five patients and cholangioscopy in three. ESWL was performed twice in each of four patients. A Strecker expandable metal stent was placed in one patient to maintain sphincterotomy patency. In one patient, two sessions of IEL and pulsed laser were carried out. Complete clearance of intrahepatic stones was achieved in four of the six subjects (66.6%) and partial clearance in two patients. No morbidity or mortality was observed. During the follow-up (mean 6.2 years; range: 2.1-16.3), only two patients had acute cholangitis at nine months and three years, respectively, after the endoscopic treatment. Both had residual intrahepatic stones left after the initial endoscopic attempt at clearance. CONCLUSION: ERCP is a necessary diagnostic procedure which should always be carried out in patients with Caroli's disease. Our experience shows that ES does not result in an increased incidence of cholangitis and that therapeutic endoscopy allows complete clearance of intrahepatic stones in the majority of patients with unresectable symptomatic Caroli's disease. Nevertheless, the oncological risk in these patients remains unchanged, and they still have an increased risk of cholangiocarcinoma. PMID- 9746167 TI - Ethanolamine injection for sclerotherapy of angiodysplasia of the colon. AB - BACKGROUND AND STUDY AIMS: Endoscopic sclerotherapy has been a very useful method for the management of bleeding vascular lesions of the gastrointestinal tract. In this report, the injection of a sclerosant agent was evaluated for the treatment of angiodysplasias of the colon. PATIENTS AND METHODS: In a prospective study in eight patients an ethanolamine solution was injected under endoscopic observation directly into 15 lesions, typically angiodysplasias of the right colon at the index colonoscopy, and into another eight de novo lesions found at subsequent examinations. The needle injector was intended to be placed very carefully into the lesion, tangentially to the mucosal surface, to avoid penetrating the bowel wall. RESULTS: Clinical follow-up showed that in six out of the eight patients (75%) no further evidence of lower intestinal hemorrhage was registered after the sclerotherapy; follow-up ranged from 22 to 36 months. Of the four patients who needed blood transfusion before the treatment because of intestinal bleeding, only one required blood transfusion after the sclerotherapy. Neither immediate nor late complications were recorded; often light bleeding occurred immediately after the injections and stopped spontaneously, except in one case which necessitated additional injection of the sclerosant. On the other hand, only one patient had light transient right lower quadrant pain after the injection, which subsided without any medication. CONCLUSIONS: Our method was shown to be feasible and safe. The success of the intralesional injection of the sclerosing agent may be predicted when changes in the mucosal surface are observed: (a) immediately after the injection sufficient sclerosant is deemed to have been injected and to the proper depth in the bowel wall, if the mucosa bulges while the solution is being injected; and (b) if a shallow ulceration is seen in an early subsequent reexamination where the treated lesion was located, allowing scar tissue produced by the healing process of the ulcer to replace the former vascular lesion. PMID- 9746168 TI - Perception and interpretation: the problem of the visible vessel. PMID- 9746169 TI - Esophageal stenting--when should metal replace plastic? PMID- 9746170 TI - Endoscopic treatment of early cancer in Japan: have we reached the limit? PMID- 9746171 TI - Severe anastomotic bleeding without a mucosal defect after partial gastrectomy. PMID- 9746172 TI - Inappropriate rise in heart rate caused by intravenous administration of trospium chloride during upper gastrointestinal endoscopy. PMID- 9746173 TI - Hemolysis due to G-6-PD deficiency induced by endoscopic sphincterotomy. PMID- 9746174 TI - Management of gastrointestinal fistulas with n-2-butyl-cyanoacrylate. PMID- 9746175 TI - Endoscopic management of a retrogastric abscess complicating laparoscopic appendectomy. PMID- 9746176 TI - New, safe and reliable method for endoscopic gastrostomy device replacement. PMID- 9746177 TI - Endosonography-guided drainage of pancreatic pseudocyst without gastric or duodenal compression. PMID- 9746178 TI - Breakage of a biliary plastic stent--a potential risk of sideholes. PMID- 9746179 TI - Isolated gallbladder tuberculosis with postoperative biliary fistula. PMID- 9746180 TI - Prevention of splenic injury during colonoscopy by positioning of the patient. PMID- 9746181 TI - Type 2 arteriovenous malformation of the sigmoid colon with unusual angiographic and characteristic histologic appearances. PMID- 9746182 TI - New developments in jewellery and dental materials. AB - This communication reviews the latest alloys introduced in the fields of jewellery and dental prostheses. For this we have scanned current patents and others to which we have access, and it is evident that, although not always correctly used, the words "antiallergic", "hypoallergic", "non-allergic", "to avoid allergies", etc., now appear frequently, indicating manufacturers' awareness of the problems that nickel can cause. On the other hand, the problems that may be associated with other sensitizing metals, such as cobalt and palladium, are not yet addressed. PMID- 9746183 TI - Oxidation of resin acids in colophony (rosin) and its implications for patch testing. AB - Commercial preparations of colophony (rosin) used for patch testing are made from unmodified rosin in pet. and may be stored for some considerable time before being used. This would be satisfactory if the composition and dermatological activity of the preparations were both reproducible and stable, but investigations by the authors have shown that the resin acids undergo progressive and substantial oxidation and that the dermatological activity of the preparations increases significantly with time. This may be a cause of inconsistent patch test results unless the composition can be stabilized. Gas liquid chromatography (GLC) analysis of a raw rosin sample and its commercial patch test preparation has shown that they both contained the same resin acids, but the concentration of the abietic type resin acids was found to be lower in the patch test preparations. The degradation of resin acids is due to their atmospheric oxidation, which may occur during the preparation and storage of the commercial rosin patch test preparation. The susceptibility of individual resin acids to atmospheric oxidation was demonstrated by analysing a sample of raw Portuguese gum rosin, which was then left exposed to air and light. Most of the resin acids were found to undergo oxidation at a rate which gradually diminished. More importantly, it is presumed that the concentration of oxidized resin acids increased correspondingly, and these have been shown to be more dermatologically active than the unoxidised resin acids. The rate of decrease of resin acid concentration was found to be in the following order: neoabietic>levopimaric and palustric>abietic>dehydroabetic acid. The pimaric type resin acids were found to be relatively inert to atmospheric oxidation when compared with the abietic type resin acids. Patch testing with the resulting partly oxidized Portuguese rosin produced positive reactions at a 35% higher frequency than the raw Portuguese rosin. The study demonstrates that the allergic potential of unmodified rosin may increase with exposure to air and light. It is therefore recommended that rosin preparations are analysed routinely as part of a quality control programme, which will enable better validation and comparison of patch test results from different dermatological centres. Since the oxidized resin acids are the main allergens in unmodified rosin, it is important that the concentration of the oxidized resin acids is kept high as well as constant in commercial patch test preparations. PMID- 9746184 TI - Proper statistical analysis of transepidermal water loss (TEWL) measurements in bioengineering studies. AB - In irritancy studies, measurement of transepidermal water loss (TEWL) is a widely used technique to assess barrier function. Using inappropriate statistical methods, however, leads to loss of information and misinterpretation of results. In this paper, we discuss some problems and pitfalls when using a suitable statistical technique for most designs in bioengineering studies, analysis of variance (ANOVA): multiple comparisons, choice of sample size and violation of statistical assumptions. For clarification of these points, a practical example will be given. Using the proposed adequate statistical methods correctly will, although accompanied by increased complexity, increase the efficiency of bioengineering studies. PMID- 9746186 TI - Occupational skin diseases: reliability and utility of the data in the various registers; the course from notification to compensation and the costs. A case study from Denmark. AB - Occupational diseases affect many people and may have serious social and economic consequences. In 1984, the National Labour Inspection Service established the Register of Occupational Diseases (ROD). The purpose of the central register was to provide information about injury-causing factors and risk groups, etc., changes in risk factors (ongoing monitoring and warning system), and to document the effects of preventive activities. However, we are dealing with several uncertain factors (i.e., whether the number of notified cases is too high or too low), and although the ROD contains a lot of information, it basically represents only notified (suspected) cases, until otherwise proven. Therefore, the utility and reliability of the data in the ROD may be questionable. The National Board of Industrial Injuries and the insurance companies represent recognized and compensated (genuine) cases, but their registers in general contain little information on variables. Thus, it is difficult to obtain exact information of occupational diseases (i.e., the real frequency and causes). What is known, is the number of cases that are notified, recognized and compensated, and the costs. Clearly, the higher the frequency of recognition, the more representative the data in the ROD of the recognized (genuine) cases. Therefore, the course from notification to recognition, and from recognition to compensation was calculated, and, for each step, the importance of skin diseases was considered. Only in the case of skin diseases, was the frequency of recognition high (2/3), and the data in the ROD were considered in more detail, and, where possible, compared with recognized and compensated cases. The various registers concurrently showed that nearly all occupational skin diseases were eczematous in nature (98%), most cases belonged to the younger age group (2/3), women (2/3) predominated over men, and the dominant type of occupational eczema was irritant (2/3). It has not been possible to get further information about exposure sources, occupations and trades from the other registers. However, considering the high frequency of recognition for skin diseases (eczemas), it is likely that the information in the ROD is also to some extent representative of the recognized (genuine) cases. As regards the importance of various disease categories, skin diseases (eczemas) ranked 1st (numerically) among both recognized and compensated cases, and were the most expensive. Therefore preventive activities are mandatory, and because of the high frequency of recognition, the data in the ROD may provide a basis for establishment of the most relevant preventive activities. For other disease categories, the frequency of recognition was low, and the utility and reliability of the data in ROD is in questionable. Therefore, in general, an improvement in the notification system is desirable, but a system that takes into account the many uncertain factors is extremely difficult to set up. Linking of the registers is in progress, and this will be useful during everyday situations. PMID- 9746185 TI - Nickel content of standard patch test materials. AB - The nickel sulfate content in standard patch test materials currently used in the US, Europe and Japan was determined by inductively-coupled plasma atomic emission spectroscopy, a state-of-the-art microanalytical method which allows nickel detection to levels of 7 ppb (microg/l). In 2 materials with a nominal concentration of 5% nickel sulfate hexahydrate in pet., the range (calculated averages of triplicate analyses of 3 different batches) was from 4.72 to 4.87% and 4.97 to 5.39%, respectively. In 1 material with a 2.5% nominal concentration in pet., the values ranged from 2.41 to 2.48%. The range seen in a 5% NiSO4 hexahydrate aq. material was 4.95 to 5.03%. The range for 2.5% NiSO4 anh. in pet. was 2.39 to 2.49%. This data suggest a significant improvement in quality control compared to our previously published data. PMID- 9746187 TI - The effect of patch duration on the elicitation of para-phenylenediamine contact allergy. AB - To study the length of exposure time required to elicit para-phenylenediamine (PPD) allergic reactions, patients known to be allergic to PPD were recruited and patch tested. A group of 7 patients were patch tested with 1% PPD in pet. for 15 min, 30 min and for 120 min. The remaining 9 patients were patch tested with 1%, 0.3%, 0.1% and 0.01% PPD for 15 min, 30 min and for 120 min each. With exposure for 120 min, 11 of 16 subjects reacted to 1% PPD and 2 of 9 reacted to 0.01%. With exposure of 15 min, 6 of 16 reacted to 1% PPD and 0 of 9 reacted to 0.01% PPD. This study showed marked inter-individual variability in eliciting a reaction to the PPD molecule on patch testing, with regard to both the exposure time and the concentration required. PMID- 9746188 TI - Experimental study of contact dermatitis due to alstroemeria in guinea pigs. PMID- 9746189 TI - Systemic contact dermatitis due to norfloxacin with a positive patch test to quinoline mix. PMID- 9746190 TI - Photodermatitis from tetrazepam. PMID- 9746191 TI - Airborne occupational contact dermatitis from ethylene oxide. PMID- 9746192 TI - Toast-makers' fingers. PMID- 9746193 TI - Allergic contact dermatitis from kojic acid. PMID- 9746194 TI - Trimethoprim-induced fixed drug eruption: positive topical provocation on previously involved and uninvolved skin. PMID- 9746195 TI - Concomitant sensitization to high and low molecular-weight heparins, heparinoid and pentosanpolysulfate. PMID- 9746196 TI - Desquamative gingivitis, sole manifestation of tosylamide/formaldehyde resin allergy. PMID- 9746198 TI - Contact dermatitis from paraphenylenediamine used as a skin paint. PMID- 9746197 TI - Contact allergy to a testosterone patch. PMID- 9746199 TI - Occupational contact allergy to nifuroxazide simulating prurigo nodularis. PMID- 9746200 TI - The repeated open application test: suggestions for a scale of evaluation. PMID- 9746201 TI - Phototoxic reaction to parsnip and UV-A sunbed. PMID- 9746202 TI - Drug eruption due to bucillamine. PMID- 9746203 TI - Allergic contact dermatitis from acrylic resin repair of windscreens. PMID- 9746204 TI - Occupational allergic contact dermatitis in a patient with a positive patch test to tin. PMID- 9746205 TI - Selenium and liver cirrhosis. AB - Effects of selenium deficiency, induced by thioacetamide, were investigated in rats. Thioacetamide (0.3 g/L) given in drinking water, as expected, caused a significant loss of selenium from the liver. It was accompanied by liver cirrhosis and a significant increase in the liver weight as well as liver to body weight ratio. A significant loss of selenium from spleen was also accompanied by an increase in its weight. Weights of lungs, testis and kidney, however, were not affected by thioacetamide and there was no change in their selenium content. Plasma levels of selenium were significantly reduced in the thioacetamide treated group. All these changes were confirmed to be due to selenium deficiency caused by thioacetamide, as supplementation with selenium reversed these changes. The mode of action of selenium is unknown but may involve anti-oxidant defense mechanisms. PMID- 9746206 TI - Induction of programmed cell death in human retinoblastoma Y79 cells by C2 ceramide. AB - C2-ceramide, a cell-permeable analogue of ceramide, induced significant, dose- and time-dependent death in human retinoblastoma Y79 cells. Dying cells strongly displayed the morphology of apoptosis as characterized by microscopic evidence of cell shrinkage, membrane blebbing, nuclear and chromatin condensation and degeneration of the nucleus into membrane-bound apoptotic bodies. Upon induction of apoptosis Y79 cells evidence early phosphatidylserine externalization, as shown by annexin V-FITC. Apoptosis was also assessed by monitoring changes in cell granularity by staining with the combined fluorescent dyes acridine orange and ethidium bromide. C2-ceramide induced these morphological changes without a concomitant production of oligonucleosomal fragments responsible for the DNA ladder and without changes in p53 protein level. Apoptosis was accompanied by accumulation of a modified Bcl-2 protein with a slower-mobility form, and by proteolytic cleavage of PARP. The effect seemed to be specific for C2-ceramide, as C2-dihydroceramide, or other amphiphilic lipid analogues, or products of ceramide hydrolysis were ineffective. The effect also depended on mRNA and protein synthesis as it was markedly inhibited by actinomycin D and cycloheximide. Sphingomyelinase and interleukin-1beta, which are known to activate the sphingomyelin turnover leading to ceramide generation, also induced apoptosis mimicking the effects of ceramide. These findings propose ceramide as an activator of the suicidal program in Y79 cells. PMID- 9746208 TI - Gene gun-mediated in vivo analysis of tissue-specific repression of gene transcription driven by the chicken ovalbumin promoter in the liver and oviduct of laying hens. AB - In order to search tissue-specific elements in the 5'-upstream promoter region, gene gun was used to transfect in vivo plasmid DNAs with varying lengths of truncated ovalbumin promoter fused to the CAT reporter gene to the oviduct and liver of laying hens. The results indicated that in the oviduct, consistently high reporter gene expression was observed irrespective of the length of the truncated ovalbumin gene promoters, whereas in the liver the ovalbumin promoter extending from -3200 to +8 bp suppressed substantially the reporter gene expression compared with consistently high gene expression obtained by the ovalbumin promoters from -2800 to +8 bp or shorter length. It was concluded, therefore, that a tissue-specific silencer-like element might reside most likely in the ovalbumin gene promoter region between -3200 and -2800 bp which represses the ovalbumin gene transcription in the liver, but not in the oviduct of laying hens. PMID- 9746207 TI - In vitro desaturation or elongation of monotrans isomers of linoleic acid by rat liver microsomes. AB - Several nutritional studies have shown the in vivo conversion of the 9c, 12t-18:2 and 9t, 12c-18:2 into long chain polyunsaturated fatty acids (PUFA) containing 20 carbons (geometrical isomers of eicosadienoic and eicosatetraenoic acids). In the present work, some in vitro studies were carried out in order to have precise information on the conversion of these two isomers. In a first set of experiments, studies were focused on the in vitro delta6 desaturation, the first regulatory step of the biosynthesis of n-6 long chain PUFA, from 9c, 12c-18:2. Rat liver microsomes were prepared and incubated under desaturation conditions with [1-14C]-9c, 12c-18:2 in presence of unlabelled 9c, 12t-, 9t, 12c- or 9t, 12t 18:2. The data show that each trans isomer induced a decrease of the delta6 desaturation of the [1-14C]-9c, 12c-18:2, but the 9c, 12t-18:2 was the most potent inhibitor (up to 63%). Rat liver microsomes were also incubated with [1 14C]-9c, 12c-18:2, [1-14C]-9c, 12t-18:2 or [1-14C]-9t, 12c-18:2 under desaturation conditions. The results indicated that 18:2 delta9c, 12t is a much better substrate for desaturase than 9t, 12c-18:2. Moreover, the conversion levels of [1-14C]-9c, 12t-18:2 was similar to what was observed for its all cis homologue, at low substrate concentration only. In a second set of experiments, in vitro elongation studies of each mono-trans 18:2 isomer and 9c, 12c-18:2 were carried out. For that purpose, rat liver microsomes were incubated with [1-14C] 9c, 12c-18:2, [1-14C]-9c, 12t-18:2 or [1-14C]-9t, 12c-18:2 underelongation conditions. The data show that [1-14C]-9t, 12c-18:2 is betterelongated than 9c, 12c-18:2 while the amount of product formed from [1-14C]-9c, 12t-18:2 was lower than was produced from the 9c, 12c-18:2. Thus, the desaturation enzymes presented a higher affinity for the 9c, 12t-18:2 whereas the elongation enzyme presented a higher affinity for the 9t, 12c-18:2. PMID- 9746209 TI - Brassica napus hsp90 can autophosphorylate and phosphorylate other protein substrates. AB - A Brassica napus cDNA encoding the 90 kDa heat shock protein, hsp90, was modified to add 6 histidines at the C-terminus and expressed in insect cells to prepare a recombinant histidine-tagged hsp90. The recombinant protein was purified over Ni2+-NTA agarose columns and its identity was confirmed by Western blotting, using a plant hsp90-specific antiserum. Incubation of purified hsp90 with [gamma 32P] ATP in the presence of Mn2+ resulted in its autophosphorylation on serine residues. The purified hsp90 could also phosphorylate other protein substrates such as histones and casein in the presence of Mn2+. Analysis of phosphorylated casein revealed that serine residues are phosphorylated by hsp90. This is the first demonstration that a cytosolic hsp90 homolog can phosphorylate other protein substrates. PMID- 9746211 TI - Modulation of the induction of ornithine decarboxylase by some opioid receptor agonists in immune cells and cardiomyocytes. AB - The ability of natural and synthetic opioids to modulate the induction of ornithine decarboxylase (ODC) was investigated in immune cells and cardiomyocytes in culture. In particular, Leu-enkephalin, which shows preference for delta receptors, enhanced ODC activity in both thymocytes and cardiomyocytes, whereas the effect of U-50488H, a synthetic kappa-selective agonist, was cell-specific. In thymocytes, U-50488H markedly inhibited the induction of the enzyme elicited by the mitogen concanavalin A (Con A) or by a combined treatment with PMA and A23187, and also reduced basal ODC activity. However the drug did not affect ODC induced by other stimuli. The inhibition of the induction of ODC activity was accompanied by a reduction of ODC mRNA level and an acceleration of ODC turnover. The action of U-50488H in thymocytes does not appear to be mediated by kappa or other classical opioid receptors lacking both stereospecificity and antagonist sensitivity, but may involve a pertussis toxin-sensitive G protein. Splenocytes also showed the ODC inhibiting effect of U-50488H, although they were less sensitive compared to thymocytes. In contrast, U-50488H enhanced ODC activity in cardiomyocytes and this effect was blocked by a specific kappa-antagonist. In conclusion, these results indicate that some opioid agonists can modulate ODC expression in non neural cells. In particular, kappa-opioid receptors may be involved in the U-50488H action in cardiomyocytes, and a distinct site, linked to inhibition of cell proliferation, may operate in immune cells. PMID- 9746210 TI - The inflammation modulatory protein (IMP) of cowpox virus drastically diminishes the tissue damage by down-regulating cellular infiltration resulting from complement activation. AB - Vaccinia virus (VV) and other pathogenic poxviruses encode for a complement control protein. The VV complement control protein or VCP, was one of the first soluble microbial proteins postulated to have an active role in the immunomodulation of the host defense. Since then, 2 other poxviruses, including variola virus and cowpox virus (CPV), were found to have corresponding proteins. Based upon earlier studies which demonstrated the role of the CPV complement control protein in modulating the specific tissue responses in BALB/c and congenic-matched C5-sufficient and C5-deficient mice, the CPV equivalent has been renamed the inflammation modulatory protein (IMP), so as to specifically reflect its function. In this study, the in vivo cellular response of mice injected with CPV or a recombinant virus lacking the IMP sequence (CPV-IMP) was examined using a connective tissue air pouch model. Microscopic examination revealed that CPV IMP caused a significant mononuclear cell infiltration into the connective tissue and adjacent dermal tissue of the skin. To characterize IMP's ability to regulate the observed cellular infiltration through both complement derived and non complement derived chemotactic factors, footpad and skin connective tissue of C3 knockout mice and footpad of MIP-1alpha knockout mice received injections of CPV and CPV-IMP. In comparison to the matched control, significantly greater footpad specific swelling response was seen in C3 -/- mice injected with CPV. This indicates an important role for C3 in poxvirus pathogenesis. However, MIP-1 alpha -/- mice injected with CPV-IMP recovered earlier than mice injected with CPV alone. This indicates that the function of IMP in vivo in mice with a complete repertoire of immune components is to limit cellular infiltration by down regulating the complement derived chemotactic analphylotoxins, thereby modulating the inflammatory response contributing to a diminished tissue pathology and preservation of viral habitat. PMID- 9746213 TI - Effects of L-carnitine on mechanical recovery of isolated rat hearts in relation to the perfusion with glucose and palmitate. AB - The purpose of this study was to investigate the effects of L-carnitine on the hemodynamic parameters of Langendorff hearts. Isolated rat hearts were perfused with various solutions containing high or low concentrations of fatty acids, additional glucose or no glucose, and L-carnitine or no L-carnitine. The most interesting part of the experiments was the behaviour of the hearts in the reperfusion period after no-flow ischemia of 20 min. The results were: (1) With glucose and high fatty acid concentrations the hearts showed an improved recovery of the left ventricular functions in the reperfusion period compared with low fatty acid concentrations. Without glucose the left ventricular pressure is much lower in the reperfusion period. (2) Addition of L-carnitine improved the recovery of the ischemically damaged hearts. This improvement is especially impressive at low fatty acid concentrations. L-carnitine addition at high fatty acid concentrations but without glucose strongly improved reperfusion behaviour. (3) The coronary flow is increased by 2 experimental conditions: (i) perfusion at low levels of fatty acids, carnitine and with glucose and (ii) high levels of fatty acids and carnitine but without glucose. These findings suggest that supplementation of L-carnitine has a beneficial effect on the isolated heart under various conditions, and possibly on specific human heart diseases. PMID- 9746212 TI - cAMP-dependent phosphorylation and hexamethylene-bis-acetamide induced dephosphorylation of p19 in murine erythroleukemia cells. AB - The objective of this study was to investigate cyclic-adenosinemonophosphate (cAMP)-dependent phosphorylation in murine erythroleukemia (MEL) cells and to identify either direct substrates of cAMP-dependent kinase or downstream effectors of cAMP dependent phosphorylation with a potential function in growth and differentiation. MEL-cells rendered deficient in cAMP-dependent protein kinase (A-kinase) activity by stable transfection with DNA encoding for either a mutant regulatory subunit or a specific peptide inhibitor of A-Kinase (PKI) are unable to differentiate normally in response to chemical inducers. We have identified by 2-D Western blotting 2 phosphorylated forms of p19, a highly conserved 18-19 kDa cytosolic protein that is frequently upregulated in transformed cells and undergoes phosphorylation in mammalian cells upon activation of several signal transduction pathways. The phosphorylation of the more acidic phosphorylated form is increased in a cAMP-dependent fashion and impaired in cells deficient in cAMP-dependent kinase (A-kinase). Treatment of MEL cells with the chemical inducer of differentiation hexamethylene-bisacetamide (HMBA) led to dephosphoryation of this phosphoform. Our data are compatible with previous observations which imply that phosphorylation of Ser 38 in p19 by p34cdc2-kinase leads to a more basic phosphoform and simultaneous phosphorylation by mitogen-activated kinase of Ser 25 in response to protein kinase C and the cAMP-dependent kinase creates the more acidic species. PMID- 9746214 TI - Role of glutathione on renal mitochondrial status in hyperoxaluria. AB - Role of glutathione on kidney mitochondrial integrity and function during stone forming process in hyperoxaluric state was investigated in male albino rats of Wistar strain. Hyperoxaluria was induced by feeding ethylene glycol (EG) in drinking water. Glutathione was depleted by administering buthionine sulfoximine (BSO), a specific inhibitor of glutathione biosynthesis. Glutathione monoester (GME) was administered for supplementing glutathione. BSO treatment alone or along with EG, depleted mitochondrial GSH by 40% and 51% respectively. Concomitantly, there was remarkable elevation in lipid peroxidation and oxidation of protein thiols. Mitochondrial oxalate binding was enhanced by 74% and 129% in BSO and BSO + EG treatment. Comparatively, EG treatment produced only a 33% increase in mitochondrial oxalate binding. Significant alteration in calcium homeostasis was seen following BSO and BSO + EG treatment. This may be due to altered mitochondrial integrity and function as evidenced from decreased activities of mitochondrial inner membrane marker enzymes, succinate dehydrogenase and cytochrome-c-oxidase and respiratory control ratio and enhanced NADH oxidation by mitochondria in these two groups. NADH oxidation (r = -0.74) and oxalate deposition in the kidney (r = -0.70) correlated negatively with mitochondrial glutathione depletion. GME supplementation restored normal level of GSH and maintained mitochondrial integrity and function, as a result of which oxalate deposition was prevented despite hyperoxaluria. These results suggest that mitochondrial dysfunction resulting from GSH depletion could be a contributing factor in the development of calcium oxalate stones. PMID- 9746215 TI - The salubrious effect of tamoxifen [correction of Tamaxifen] on serum marker enzymes, glycoproteins, and lysosomal enzymes level in breast cancer woman. AB - Tumour markers correlate strongly with prognosis based on tumour burden and surgical resectability. If chemotherapy is extremely effective in certain stage of the disease, the sensitive marker may be of great use in monitoring disease response and drug treatment. Hence, this study was launched to evaluate the changes in tumour marker enzymes like lactate dehydrogenase (LDH), glutamate oxaloacetate transaminase (SGOT), glutamate pyruvate transaminase (SGPT), alkaline phosphatase, and acid phosphatase in before and after 3 and 6 months tamoxifen treated breast cancer patients. In addition, the changes in serum glycoproteins viz., hexose, hexosamine, and sialic acid and lysosomal enzymes such as N-acetyl-beta-D-glucosaminidase, beta-D-galactosidase, and beta-D glucuronidase were analysed in these patients. These values were compared with their age matched healthy control subjects. At 6 months evaluation, the tamoxifen treated postmenopausal breast cancer women showed a statistically significant decreased (p < 0.001, 0.05 respectively) levels of LDH, SGOT, SGPT, alkaline and acid phosphatases than their baseline values. Similarly, the levels of hexose, hexosamine, and sialic acid and N-acetyl-beta-D-glucosaminidase, beta-D galactosidase, and beta-D-glucuronidase were decreased significantly (p < 0.001) in tamoxifen received postmenopausal women. The result of this study suggested that tamoxifen potentially retard the metastasis of breast cancer as well as the bone demineralisation in postmenopausal breast cancer women. Thus, tamoxifen may also have its antitumour activity through its beneficial effects on tumour marker enzymes and serum proteins in breast cancer women. PMID- 9746218 TI - Isolation, purification and partial characterization of chloragocytes from the earthworm species Lumbricus terrestris. AB - Chloragocytes were isolated from the earthworm species Lumbricus terrestris. After mechanical dissociation and sedimentation through Percoll, a highly purified fraction of viable chloragocytes was obtained. The isolated chloragocytes accumulated the vital dye neutral red and reduced the tetrazolium dye MTT, thereby indicating cellular integrity. Time of flight flow cytometric analyses revealed a main population of large and highly granulated cells in the 30-33 microm size range. Hydrolase measurements showed that beta-D-N-acetyl glucosaminidase and acid phosphatase exhibited the highest activities (146.6 and 24.9 mU/mg of protein, respectively), possibly indicating a major role for these 2 hydrolases in the physiological function of chloragocytes. In contrast, other acid hydrolases such as beta-D-galactosidase and beta-D-glucuronidase had specific activities of respectively 26 and 182 times lower than the glucosaminidase. The specific activity of the membrane-bound alkaline phosphatase was comparable to that of its acid counterpart (18.9 vs. 24.9 mU/mg of protein, respectively) and this level of activity may show an important trans-membrane activity in chloragocytes. The cytoplasmic and mitochondrial enzyme isocitrate dehydrogenase had a level of activity comparable to that of the exclusively cytoplasmic enzyme lactate dehydrogenase (6.6 vs. 8.1 mIU/mg of protein, respectively). When L. terrestris chloragocyte homogenates were separated on Percoll, results showed that hydrolases and dehydrogenases were mainly associated with the lighter materials that remained above the Percoll layer. Nonetheless, the detection of significant proportions (15-25%) of the total recovered activity of acid phosphatase and beta-galactosidase in the enriched chloragosome fraction supports the notion that some chloragosomes may be 'lysosome-like' organelles. PMID- 9746216 TI - cAMP-dependent phosphorylation sites and macroscopic activity of recombinant cardiac L-type calcium channels. AB - The involvement of cAMP-dependent phosphorylation sites in establishing the basal activity of cardiac L-type Ca2+ channels was studied in HEK 293 cells transiently cotransfected with mutants of the human cardiac alpha1 and accessory subunits. Systematic individual or combined elimination of high consensus protein kinase A (PKA) sites, by serine to alanine substitutions at the amino and carboxyl termini of the alpha1 subunit, resulted in Ca2+ channel currents indistinguishable from those of wild type channels. Dihydropyridine (DHP)-binding characteristics were also unaltered. To explore the possible involvement of nonconsensus sites, deletion mutants were used. Carboxyl-terminal truncations of the alpha1 subunit distal to residue 1597 resulted in increased channel expression and current amplitudes. Modulation of PKA activity in cells transfected with the wild type channel or any of the mutants did not alter Ca2+ channel functions suggesting that cardiac Ca2+ channels expressed in these cells behave, in terms of lack of PKA control, like Ca2+ channels of smooth muscle cells. PMID- 9746217 TI - Decreased phosphorylation of a low molecular weight protein by cGMP-dependent protein kinase in variant HL-60 cells resistant to nitric oxide- and cGMP-induced differentiation. AB - We previously described the isolation of a variant subline of HL-60 cells that does not differentiate in response to nitric oxide (NO)-generating agents or to cGMP analogs. The variant cells have normal guanylate cyclase activity and normal NO-induced increases in the intracellular cGMP concentration. We now show that the variant cells have normal cGMP-dependent protein kinase (G-kinase) activity, both by an in vitro and in vivo assay, and using two-dimensional gel electrophoresis we have identified six G-kinase substrates in the parental cells. Of these six proteins, we found considerably less phosphorylation of one of the proteins in the variant cells than in parental cells, both in vitro and in intact cells, and by 35S-methionine/35S-cysteine incorporation we found much less of this protein in the variant cells than in parental cells. The protein is a shared substrate of cAMP-dependent protein kinase (A-kinase); since cAMP analogs still induce differentiation of the variant cells, it appears that the NO/cGMP/G-kinase and cAMP/A-kinase signal transduction pathways share some but not all of the same target proteins in inducing differentiation of HL-60 cells. PMID- 9746219 TI - Purification and characterization of UDP-GalNAc:polypeptide N acetylgalactosaminyl transferase from swine trachea epithelium. AB - UDP-GalNac: polypeptide N-acetylgalactosaminyltransferase from swine trachea epithelium was purified to homogeneity by procedures which included affinity chromatography on Sepharose 4B columns containing bound deglycosylated Cowper's gland mucin. The enzyme, purified 12,000-fold from microsomes with a yield of 40%, showed only a single band on dodecyl sulfate polyacrylamide gel electrophoresis. The homogenous enzyme has an apparent molecular mass of 70,000 Da, as determined by gel electrophoresis or gel filtration. The transferase has a broad pH optimum between 6.7-7.8 with maximal activity at pH 7.2, and required Mn2+ for activity with maximal activity at 5-7.5 mM. Higher concentrations of Mn2+, inhibited the enzyme. The purified transferase was specific for UDPGalNAc and glycosylated both threonine and serine residues in tryptic peptides prepared from deglycosylated Cowper's gland and swine and human trachea mucins. The apparent Km of the transferase for UDPGalNAc was 6.3 microM, and the Km values for deglycosylated Cowper's gland and human and swine trachea mucins were 0.83, 1.12 and 0.94 mg/ml, respectively. The Vmax of the purified enzyme was 2.1 micromol/min/mg with deglycosylated Cowper's gland mucin, as the glycosyl acceptor. However, the activities with peptides prepared from deglycosylated mucins by limited acid hydrolysis were 20-fold greater than the intact glycoprotein under identical conditions. The deglycosylated mucins and larger peptides aggregated with time of storage and precipitated from solution. Aggregation was accompanied by a corresponding loss of enzymatic activity even after dispersion of the aggregate by sonication. The deglycosylated mucins which were prepared by chemical treatment and periodate oxidation still contained about 20% of the N-acetylgalactosamine present in the intact mucin. When this residual amino sugar was removed by periodate oxidation the completely deglycosylated mucins became very poor substrates for the purified transferase. Data obtained in the current study indicate that the accessibility of serine and threonine in the polypeptide chains of mucin glycoproteins significantly influences the rate of glycosylation of these amino acids. The best substrates and affinity ligand for the enzyme were fragments of incompletely deglycosylated mucin polypeptide chains. PMID- 9746220 TI - Lithostathine messenger RNA expression in different types of chronic pancreatitis. AB - Lithostathine may play a physiological role in preventing the precipitation of excess calcium in the pancreatic juice. The hypothesis has been advanced that in chronic calcifying pancreatitis the abnormal biosynthesis of lithostathine might be the original defect to which genetic proneness to the disease may be ascribed. The aim of the present work was to study lithostathine messenger RNA expression in the pancreas of patients with different types of pancreatitis. Lithostathine and chymotrypsinogen mRNA were determined in surgical specimens obtained from the pancreases of the following subjects: (a) 13 patients with chronic alcoholic pancreatitis (84.6% calcified); (b) 4 patients with chronic hereditary pancreatitis (all calcified); (c) 6 patients with chronic obstructive pancreatitis (4 calcified); and (d) 27 subjects suffering from pancreatic cancer. Significantly lower concentrations of both mRNAs were found in the pancreases of chronic pancreatitis patients than in non-cancerous tissue from pancreatic cancer subjects. However, about 70% of the pancreatic cancer subjects showed lithostathine and chymotrypsinogen mRNA levels comparable to those of chronic pancreatitis patients. These results indicate that the decrease in the level of mRNA is not specific to lithostathine and it is unrelated to the presence of pancreatic stones. PMID- 9746221 TI - Alteration in calcium content and Ca2+-ATPase activity in the liver nuclei of rats orally administered carbon tetrachloride. AB - The alteration in calcium transport in the liver nuclei of rats orally administered carbon tetrachloride (CCl4) was investigated. Rats received a single oral administration of CCl4 (5, 10, and 25%, 1.0 ml/100 g body weight), and 5, 24 and 48 h later the animals were sacrificed. The administration of CCl4 (25%) caused a remarkable elevation of calcium content in the liver tissues and the nuclei of rats. Liver nuclear Ca2+-ATPase activity was markedly decreased by CCl4 (25%) administration. The presence of dibutyryl cyclic AMP(10(-4) and 10(-3) M) or inositol 1,4,5-trisphosphate (10(-6) and 10(-5) M) in the enzyme reaction mixture caused a significant decrease in Ca2+-ATPase activity in the liver nuclei obtained from normal rat, while the enzyme activity was significantly increased by calmodulin (1.0 and 2.0 microg/ml). These signaling factor's effects were completely impaired in the liver nuclei obtained from CCl4 (25%)-administered rats. DNA fragmentation in the liver nuclei obtained from CCl4-administered rats was significantly decreased by the presence of EGTA (2 mM) in the reaction mixture, suggesting that the endogenous calcium activates nuclear DNA fragmentation. The present study demonstrates that calcium transport system in the liver nuclei is impaired by liver injury with CCl4 administration in rats. PMID- 9746222 TI - Hypothyroidism-evoked shifts in hippocampal adrenergic receptors: implications to ischemia-induced hippocampal damage. AB - Hypothyroidism was induced in a group of male Fischer 344 rats by administration of 0.05% propylthiouracil (PTU) in the drinking water for 12 weeks. Control rats were not treated. Plasma levels of thyroid hormones indicated that PTU treatment had produced severe thyroid hormone deficiency. In PTU-treated rats compared to control rats, levels of total T3 and total T4 were reduced 54.5% and 53.7%; while levels of free T3 and free T4 were reduced 87.1% and 96.5%. Functional hypothyroidism was demonstrated by: (i) a 49.1% decrease in hepatic plasma membrane alpha1-adrenergic receptor binding, and (ii) a 11.2-fold increase in hepatic gamma-glutamyltranspeptidase activity; relative to the expression of these parameters in control rats. Membranes were isolated from hippocampi of control, PTU-induced hypothyroid and thyroxine-replaced rats and specific adrenergic receptor binding determined by radioligand binding techniques. Hypothyroidism resulted in a shift in the balance of alpha1 and beta2 adrenergic receptor binding by evoking: an increase in alpha1-adrenergic receptor binding to 1.57-fold of control levels; and, a decrease in beta2-adrenergic receptor binding to 64% of control levels. Thyroid hormone replacement carried out in PTU-treated hypothyroid rats at 30 microg/kg s.c. per day for the last 3 days of the 12 week PTU-treatment protocol, which reversed physical and functional hypothyroidism, reversed the observed changes in hippocampal adrenergic receptor binding, indicating them to be thyroid hormone, and not PTU, -dependent. This receptor shift evoked by hypothyroidism may, in part, explain the protective effect of hypothyroidism on ischemia-induced hippocampal damage by favoring inhibitory input and limiting excitotoxic input by catecholamines. PMID- 9746223 TI - In vitro induction of nitric oxide by fructose-1,6-diphosphate in the cardiovascular system of rats. AB - Nitric oxide (NO) functions as a cellular messenger in a number of organs and cell systems in the cardiovascular system (CVS); it is a significant determinant of basal vascular tone and regulates myocardial contractility and platelet aggregation. The present study focused upon understanding the in vitro effects of fructose-1,6-diphosphate (FDP) on the rat cellular NO pathway. The iNOS activity was measured by monitoring the formation of (3H)-citrulline in 50,000 g soluble fractions of crude homogenates of endothelial (ET) and smooth muscle cells (SMC) from the arteries of rats, and macrophages (MAC) and lymphocytes (LYM) from rat blood. FDP in concentrations of 10-1000 microM stimulated rat cellular iNOS activity in a concentration-dependent manner. FDP-stimulated rat cellular iNOS was found to be completely reversed by 5 microM concentration of NG-monomethyl-L arginine (L-NMMA), the potent mammalian NOS inhibitor. These studies demonstrated that FDP may induce the formation of NO by stimulating rat cardiovascular iNOS activity. PMID- 9746224 TI - In vivo tissue specific modulation of rat insulin receptor gene expression in an experimental model of mineralocorticoid excess. AB - Insulin receptor (IR) gene expression at the mRNA level was investigated in hindlimb skeletal muscle, epididymal adipose tissue and in the liver of rats exposed to prolonged in vivo administration of deoxycorticosterone acetate (DOCA). Following treatment, plasma insulin levels were reduced while glucose levels increased compared to values in control rats. DOCA-treated animals showed an increase in blood pressure and a reduction in body weight. This treatment also induced hypokalemia and decreased plasma protein levels. Sodium levels were unaffected. Moreover, no differences in DNA and protein content or in the indicator of cell size (protein/DNA) were observed in the skeletal muscle or adipose tissue of animals. In contrast, there was a clear increase in the protein and DNA contents of the liver with no change in the indicator of cell size. Northern blot assays revealed 2 major IR mRNA species of approximately 9.5 and 7.5 Kb in the 3 tissues from control animals. DOCA treatment induced no change in the levels of either RNA species in skeletal muscle. However, a decrease of approximately 22% was detected in the levels of both species in adipose tissue whereas the liver showed an increase of 64%. These results provide the first evidence for an in vivo tissue-specific modulation of IR mRNA levels under experimental conditions of mineralocorticoid excess. PMID- 9746225 TI - Changes in cardiac electrophysiology, morphology, tissue biochemistry and vascular reactions in glutathione depleted animals. AB - The effects of acute and chronic glutathione depletion (single i.p. injection of 3 mmol/kg L-buthionine-S,R-sulphoximine and 2 mmol/kg for 4 days) on heart action potential (AP) characteristics, electronmicroscopy, cytochemistry and biochemistry and vascular contractility and nitric oxide-mediated relaxation were studied in rats and guinea pigs. In guinea pig cardiac preparations both acute and chronic glutathione depletion caused a significant decrease of maximum rate of rise of depolarization phase and duration of action potential AP(APD) at 25, 50, and 90% of repolarization but did not modify the other AP parameters. The contractile responses of helically cut aortic strips to norepinephrine were not altered by chronic glutathione depletion but the relaxing responses of precontracted preparations to acetylcholine were significantly reduced both in rats and guinea pigs. Morphologically there were indications of permeability changes, intracellular and interstitial edema and myofilament damage in the myocardium. There was also a decrease in cytochromoxydase and succinyl dehydrogenase activities both in rats and guinea pigs. The present data suggest that glutathione depletion may influence the Na+ and K+ channel activities, causes morphological and biochemical changes in cardiac preparations and may interfere with nitric oxide generation or its action in aortic strips. PMID- 9746226 TI - Differential distribution and metabolism of arachidonic acid and docosahexaenoic acid by human placental choriocarcinoma (BeWo) cells. AB - The time course of incorporation of [14C]arachidonic acid and [3H]docosahexaenoic acid into various lipid fractions in placental choriocarcinoma (BeWo) cells was investigated. BeWo cells were found to rapidly incorporate exogenous [14C]arachidonic acid and [3H] docosahexaenoic acid into the total cellular lipid pool. The extent of docosahexaenoic acid esterification was more rapid than for arachidonic acid, although this difference abated with time to leave only a small percentage of the fatty acids in their unesterified form. Furthermore, uptake was found to be saturable. In the cellular lipids these fatty acids were mainly esterified into the phospholipid (PL) and the triacyglycerol (TAG) fractions. Smaller amounts were also detected in the diacylglycerol and cholesterol ester fractions. Almost 60% of the total amount of [3H]Docosahexaenoic acid taken up by the cells was esterified into TAG whereas 37% was in PL fractions. For arachidonic acid the reverse was true, 60% of the total uptake was incorporated into PL fractions whereas less than 35% was in TAG. Marked differences were also found in the distribution of the fatty acids into individual phospholipid classes. The higher incorporation of docosahexaenoic acid and arachidonic acid was found in PC and PE, respectively. The greater cellular uptake of docosahexaenoic acid and its preferential incorporation in TAG suggests that both uptake and transport modes of this fatty acid by the placenta to fetus is different from that of arachidonic acid. PMID- 9746227 TI - Rapid detection of poly(ADP-ribose) polymerase by enzyme-linked immunosorbent assay during its purification and improvement of its purification. AB - We report a new detection method for the purification of poly(ADP-ribose) polymerase (PARP). PARP purification generates many fractions in which PARP is usually detected by a time consuming activity assay. The development of a new method was also needed in order to decrease the utilization of radioactivity. This new method, based on an enzyme-linked immunosorbent assay (ELISA), is very rapid, sensitive, and avoids most radioactivity. Moreover, to illustrate this method, a new matrix was used, the Heparin Sepharose. This matrix was chosen for its affinity for the DNA binding proteins and because it allows the separation of whole PARP from its proteolytic fragments. PMID- 9746228 TI - Heat stress pretreatment mitigates postischemic arachidonic acid accumulation in rat heart. AB - Heat stress pretreatment of the heart is known to protect this organ against an ischemic/reperfusion insult 24 h later. Degradation of membrane phospholipids resulting in tissue accumulation of polyunsaturated fatty acids, such as arachidonic acid, is thought to play an important role in the multifactorial process of ischemia/reperfusion-induced damage. The present study was conducted to test the hypothesis that heat stress mitigates the postischemic accumulation of arachidonic acid in myocardial tissue, as a sign of enhanced membrane phospholipid degradation. The experiments were performed on hearts isolated from rats either 24 h after total body heat treatment (42 degrees C for 15 min) or 24 h after sham treatment (control). Hearts were made ischemic for 45 min and reperfused for another 45 min. Heat pretreatment resulted in a significant improvement of postischemic hemodynamic performance of the isolated rat hearts. The release of creatine kinase was reduced from 30 +/- 14 (control group) to 17 +/- 5 units/g wet wt per 45 min (heat-pretreated group) (p < or = 0.05). Moreover, the tissue content of the inducible heat stress protein HSP70 was found to be increased 3-fold 24 h after heat treatment. Preischemic tissue levels of arachidonic acid did not differ between heat-pretreated and control hearts. The postischemic ventricular content of arachidonic acid was found to be significantly reduced in heat-pretreated hearts compared to sham-treated controls (6.6 +/- 3.3. vs. 17.8 +/- 12.0 nmol/g wet wt). The findings suggest that mitigation of membrane phospholipid degradation is a potential mechanism of heat stress-mediated protection against the deleterious effects of ischemia and reperfusion on cardiac cells. PMID- 9746229 TI - On-line coupling of polymerase chain reaction and capillary electrophoresis for automatic DNA typing and HIV-1 diagnosis. AB - We demonstrate an integrated on-line system with a fused-silica capillary as the microreactor for PCR and capillary gel electrophoresis with laser-induced fluorescence detection for DNA typing and disease diagnosis. Two applications have been investigated: the four short tandem repeat (STR) loci vWA, THO1, TPOX and CSF1PO (CTTv) for DNA typing, and DNA probe for human immunodeficiency virus (HIV-1) diagnosis. The CTTv are important loci in forensic and genetic linkage analysis. The PCR technique is a powerful tool in HIV research because it can detect the presence of the virus before any antibody response in the infected person. Thus it is important for early diagnosis. Multiplexed PCR in a fused silica capillary, on-line injection, DNA denaturation and calibration based on a standard ladder have been successfully combined. Also, on-line liquid flow management, DNA separation and detection have been completely integrated. PMID- 9746230 TI - Study of phosphorothioate-modified oligonucleotide resistance to 3'-exonuclease using capillary electrophoresis. AB - The effect of phosphorothioate (PS) internucleotide linkages on the stability of phosphodiester oligodeoxyribonucleotides (ODNs) was investigated using 25-mer ODNs containing single or multiple PS backbone modifications. The in vitro stability of the oligomers was measured both in 3'-exonuclease solution and in plasma. For the separation of ODNs, capillary electrophoresis with a replaceable polymer separation matrix was used. As expected, DNA fragments with PS linkages at the 3'-end were found to be more resistant to 3'-exonuclease hydrolysis. Also increasing exonuclease resistance was the non-specific adsorption of phosphorothioate ODNs to enzyme. PMID- 9746231 TI - High-resolution capillary electrophoretic separation of supercoiled plasmid DNAs and their conformers in dilute hydroxypropylmethyl cellulose solutions containing no intercalating agent. AB - The three conformers of plasmid pBR322, linear, supercoiled and nicked circular forms, were separated by capillary electrophoresis (CE) in 0.1% hydroxypropylmethyl cellulose (HPMC) solution in the absence of intercalating agents and the migration order was confirmed by co-migration of enzymatically prepared corresponding DNAs. The previously observed broad peaks of supercoiled DNAs in CE are results of unresolved peaks of topoisomers which differ only in the degrees of twisting. We have demonstrated the separation of an artificial topoisomer ladder made from pBR322 and topoisomerase I. The population of topoisomers of a supercoiled DNA is dependent on sample matrices and separation conditions. PMID- 9746232 TI - On-line concentration of neutral analytes for micellar electrokinetic chromatography. VI. Stacking using reverse migrating micelles and a water plug. AB - Utility of a second enhanced field zone (water zone) is investigated for the on line concentration of neutral analytes in micellar electrokinetic chromatography. Micellar solutions of sodium dodecyl sulfate prepared in acidic phosphate buffers are used as separation and sample solutions. Prior to long hydrodynamic injection of samples prepared in a low conductivity matrix, a long water plug is hydrodynamically injected to provide a second enhanced field zone. Practical and some fundamental considerations are presented. The technique is selective towards hydrophobic analytes. Notable detector response improvements (>100-fold) for several analytes are observed experimentally. PMID- 9746233 TI - Sample enrichment in a single levitated droplet for capillary electrophoresis. AB - This paper describes sample enrichment in a single levitated droplet for capillary electrophoresis (CE) analysis. The droplet was trapped in an acoustical field. The minute sample volumes needed for the enrichment procedure were precisely handled using a piezoelectric flow-through liquid microdispenser. Droplets with a volume of 65 pl were ejected from the device at a repetition rate ranging from one single droplet up to several hundreds per second. By counting the number of droplets ejected and accumulated in the levitated drop the sample volume was controlled. Through solvent evaporation the analytes were enriched in the diminishing droplet. The droplet was then injected into a CE capillary and the analytes, dansyl-Gly and dansyl-Val dissolved in ethanol, were separated in a 100 mM borate buffer (pH 9.0) utilising UV-absorption detection at 200 nm near the capillary outlet. Enrichment of 36000 sample droplets (2.3 microl) through solvent evaporation in the levitated drop resulted in a concentration limit of detection (CLOD) of 15 nM for the dansylated amino acids as compared to a CLOD of 2.5 microM which was achieved using standard hydrodynamic injection without preconcentration. PMID- 9746234 TI - Capillary electrophoresis with laser-induced native fluorescence detection for profiling body fluids. AB - Laser-induced native fluorescence detection with a KrF excimer laser (lambda=248 nm) was used to investigate the capillary electrophoretic (CE) profiles of human urine, saliva and serum without the need for sample derivatization. All separations were carried out in sodium phosphate and/or sodium tetraborate buffers at alkaline pH in a 50-microm I.D. capillary. Sodium dodecyl sulfate was added to the buffer for micellar electrokinetic chromatography (MEKC) analysis of human urine. Although inherently a pulsed source, the KrF excimer laser was operated at a high pulse repetition rate of 553, 1001 or 2009 Hz to simulate a continuous wave excitation source. Detection limits were found to vary with pulse rate, as expected, in proportion to average excitation power. The following detection limits (3sigma) were determined in free solution CE: tryptophan, 4 nM; conalbumin, 10 nM; alpha-lactalbumin, 30 nM. Detection limits for indole-based compounds and catecholamine urinary metabolites under MEKC separation conditions were in the range 7-170 nM. PMID- 9746235 TI - Competitive immunoassay for cyclosporine using capillary electrophoresis with laser induced fluorescence polarization detection. AB - Frequent monitoring of immunosuppressive drug cyclosporine A (CsA) in blood samples of tissue transplant patients is required in clinical practice because of the narrow therapeutic range between the immunosuppressive effect and the toxic effect of this drug. We describe a competitive immunoassay capillary electrophoresis (CE) with laser induced fluorescence polarization detection method, which is rapid and sensitive for the determination of CsA. The method is based on the competitive immunochemical reaction between the analyte and fluorescent hapten (CsA*) with the antibody, CE separation of the antibody bound and free fluorescent CsA*, followed by the laser induced fluorescence polarization detection (LIFP) of the fluorescent species. The method detection limit is governed by the stability of the antibody-CsA* complex rather than by the detector noise. The use of post-column sheath flow cuvette LIFP detection resulted in excellent detection limit, typically 0.9 nM (or 9.10(-19) mol for 1 nl injection) of CsA. CsA in whole blood samples from organ transplant patients were measured and results agreed well with those obtained by using a standard fluorescence polarization immunoassay. Each determination took less than 3 min. The CsA metabolites AM9 and AM19 were also determined by using this technique, and their cross-reactivities with the antibody were 13% and 2%, respectively. PMID- 9746236 TI - Determination of chiral pharmaceutical compounds, terbutaline, ketamine and propranolol, by on-line capillary electrophoresis-electrospray ionization mass spectrometry. AB - On-line capillary electrophoresis-electrospray ionization mass spectrometry (CE ESI-MS) has been employed for the determination of racemic mixtures of the chiral drugs, terbutaline, ketamine, and propranolol. Separation of the different chiral forms has been achieved by introducing cyclodextrins (CDs), which act as chiral selectors, into the CE operating electrolytes. Cyclodextrins function as chiral selectors in CE because of their ability to form host-guest complexes (inclusion complexes) of varying stability with an array of chiral drugs and other compounds. Derivatized forms of beta-CD (i.e., dimethyl-beta-cyclodextrin and hydroxypropyl-beta-cyclodextrin) were used in this study due to their higher solubilities in the aqueous methanolic operating electrolyte than native beta-CD. Addition of minor quantities of methanol to the aqueous-based CE operating electrolytes improved the stability of electrospray ionization conditions and further enhanced CE resolution of the enantiomeric pairs relative to purely aqueous systems. Introduction of the CDs into the CE operating electrolytes caused suppression of analyte signals in ESI-MS, and the dependence of analyte signal intensities on the solution concentrations of the derivatized beta-CDs was examined. Under optimized conditions, the different enantiomeric forms of the compounds under investigation were successfully separated and detected by CE-ESI MS. PMID- 9746237 TI - Determination of dimethylamine and other low-molecular-mass amines using capillary electrophoresis with laser-induced fluorescence detection. AB - The potential of capillary electrophoresis (CE) with laser-induced fluorescence (LIF) detection for the separation and determination of dimethylamine (DMA) and other low-molecular-mass amines involving precolumn derivatization with fluorescein isothiocyanate isomer I (FITC) was investigated. Different variables that affect derivatization (pH, FITC concentration, reaction time and temperature) and separation (buffer concentration, addition of various organic modifiers, applied voltage and length of capillary) were studied. The linearity, reproducibility and reliability of the method were evaluated. The estimated instrumental detection limit for a 2-s pressure injection of the FITC-DMA derivative was 50 pg/ml (10(-9) M), using LIF detection with excitation and emission wavelengths of 488 nm and 520 nm, respectively. However, for practical reasons, a minimum of 5 ng/ml DMA should be subjected to the derivatization. The applicability of the described method to the extract of atmospheric aerosol samples was demonstrated. PMID- 9746238 TI - Micellar electrokinetic chromatography for the determination of urinary desmosine and isodesmosine in patients affected by chronic obstructive pulmonary disease. AB - The presence in urine of desmosine (DES) and isodesmosine (IDES), two crosslinked amino acids unique to the elastic fiber network, can be used as a specific indicator of degradation of mature elastin. Compared to methodologies so far available, the capillary electrophoretic technique reported here seems to be suitable and convenient for determining desmosines in urine of patients affected by chronic obstructive pulmonary disease (COPD). By using 35 mM sodium tetraborate pH 9.3 containing 65 mM SDS as the background electrolyte, the peaks of DES and IDES could be detected in hydrolyzed urine samples from controls and patients. Owing to the simultaneous determination of endogenous urinary creatinine used as appropriate internal standard, the amount of these amino acids could be accurately quantified. The results obtained were of the same order of magnitude as the data already reported in the literature for COPD patients. Thus micellar electrokinetic chromatography (MEKC) may be considered as a reliable technique for studying the turnover of the elastic fiber in clinical conditions. PMID- 9746239 TI - Separation of a range of cations by nonaqueous capillary electrophoresis using indirect and direct detection. AB - The use of nonaqueous media and indirect detection is reported for the separation and detection of a range of small cations. The novel applications involved separation of a range of metal ions, small nonchromophoric amines, cationic ion pair reagents and cationic surfactants. Separations were achieved using acidified methanol containing imidazole as the UV co-ion for indirect detection. The methods produced different selectivity compared to aqueous methods using acidified aqueous imidazole solutions. Advantages of the methods include speed of analysis and prevention of sample micellerisation. The methods were shown to be quantitative and reproducible by their application to the determination of Tris content. PMID- 9746240 TI - Novel electrolyte for the analysis of cations in low explosive residue by capillary electrophoresis. AB - A novel electrolyte has been developed for the effective separation by capillary electrophoresis of cations detected in low explosive residue. This electrolyte, with a pH of 4.4, employs 17.5 mM alpha-hydroxyisobutyric acid (HIBA) as the complexing agent, 6 mM imidazole as the ultraviolet visualization agent, 4 mM 18 crown-6 ether as a modifier to enhance the selectivity of the inorganic cations, and 5% (v/v) acetonitrile as an organic additive. Studies which assessed the value of the addition of 18-crown-6 and acetonitrile demonstrated conclusively that both were required in order to achieve unambiguous baseline separation of ammonium, potassium and monomethylammonium ions. The major advantages of the use of this electrolyte are a total run time of less than 7 min and symmetrical peak shapes. Validation on a series of preblast and postblast explosives materials determined that this procedure is reliable and robust. PMID- 9746241 TI - Automated sample introduction for an imaged capillary isoelectric focusing instrument via high-performance liquid chromatography sampling devices. AB - Sample introduction of an imaged capillary isoelectric focusing (cIEF) instrument is fully automated by using commercially available high-performance liquid chromatography (HPLC) injection valves and autosamplers. Sample carryover can be controlled to under 1% when the valve and separation column are washed for 1 min between sample runs. The standard deviation of peak areas for 20 injections is 3.5%, which includes deviations created by the absorption imaging detector and the isoelectric focusing process inside the 75 microm I.D. column. Sample throughput is up to 10 samples per hour. The instrument has been applied to fast analysis of many proteins including monoclonal antibodies. PMID- 9746242 TI - Rapid stable isotope dilution analysis of very-long-chain fatty acids, pristanic acid and phytanic acid using gas chromatography-electron impact mass spectrometry. AB - A common feature of most peroxisomal disorders is the accumulation of very-long chain fatty acids (VLCFAs) and/or pristanic and phytanic acid in plasma. Previously described methods utilizing either gas chromatography alone or gas chromatography-mass spectrometry are, in general, time-consuming and unable to analyze VLCFAs, pristanic and phytanic acid within a single analysis. We describe a simple, reproducible and rapid method using gas chromatography/mass spectrometry with deuterated internal standards. The method was evaluated by analysing 30 control samples and samples from 35 patients with defined peroxisomal disorders and showed good discrimination between controls and patients. This method is suitable for routine screening for peroxisomal disorders. PMID- 9746243 TI - Comparison of detection methods for liquid chromatographic determination of 3 nitro-L-tyrosine. AB - A liquid chromatographic method has been developed for the determination of 3 nitro-L-tyrosine. Different detection methods, including UV, oxidative and redox electrochemistry, and postcolumn photolysis followed by electrochemical detection, have been optimized and compared in terms of analysis time, detection limit and dynamic range. It was demonstrated that liquid chromatography with postcolumn photolysis followed by electrochemical detection is the most effective method, with an analysis time of 5 min, detection limit of 0.01 pmol, and a linear dynamic range from 2 nM to 100 microM. PMID- 9746244 TI - Simple purification procedure for human prostatic kallikrein hK2 in its active form. AB - Kallikrein hK2 is a new potential marker of prostate cancer. It is the last member of the human kallikrein gene family to be isolated. We propose a simple purification procedure permitting us to obtain the active form of hK2 starting from human seminal plasma and using commonly available chromatography matrices. In contrast to recently published papers, this procedure is carried out without any immunoaffinity chromatography step and without the need for any antibody to follow the purification. Furthermore, it does not require any recombinant DNA technology nor sophisticated instruments. PMID- 9746245 TI - Protein analysis by membrane preconcentration-capillary electrophoresis: systematic evaluation of parameters affecting preconcentration and separation. AB - Fast and efficient analysis of proteins in physiological fluids is of great interest to researchers and clinicians alike. Capillary electrophoresis (CE) has proven to be a potentially valuable tool for the separation of proteins in specimens. However, a generally acknowledged drawback of this technique is the limited sample volumes which can be loaded onto the CE capillary which results in a poor concentration limit of detection. In addition, matrix components in samples may also interfere with separation and detection of analytes. Membrane preconcentration-CE (mPC-CE) has proved to be effective in overcoming these problems. In this report, we describe the systematic evaluation of parameters affecting on-line preconcentration/clean-up and separation of protein mixtures by mPC-CE. Method development was carried out with a standard mixture of proteins (lysozyme, myoglobin, carbonic anhydrase, and human serum albumin). First, using MALDI-TOF-MS, membrane materials with cation-exchange (R-SO3H) or hydrophobic (C2, C8, C18, SDB) characteristics were evaluated for their potential to retain proteins in mPC cartridges. Hydrophobic membranes were found most suitable for this application. Next, all mPC-CE analysis of protein samples were performed in polybrene coated capillaries and parameters affecting sample loading, washing and elution, such as the composition and volume of the elution solvent were investigated. Furthermore, to achieve optimal mPC-CE performance for the separation of protein mixtures parameters affecting postelution focusing and electrophoresis, including the composition of the background electrolyte and a trailing stacking buffer were varied. Optimal conditions for mPC-CE analysis of proteins using a C2 impregnated membrane preconcentration (mPC) cartridge were achieved with a background electrolyte of 5% acetic acid and 2 mM ammonium acetate, 60 nl of 80% acetonitrile in H2O as an elution solvent, and 60 nl of 0.5% ammonium hydroxide as a trailing stacking buffer. The developed method was used successfully to separate proteins in aqueous humor, which contains numerous proteins in a complex matrix of salts. PMID- 9746246 TI - Biological monitoring of dinitrotoluene by gas chromatographic-mass spectrometric analysis of 2,4-dinitrobenzoic acid in human urine. AB - The method of analysis described permits the determination of 2,4-dinitrobenzoic acid down to the lower microg l(-1) range in the urine of persons exposed to dinitrotoluene. 2,4-Dinitrobenzoic acid is the main metabolite of 2,4 dinitrotoluene and technical dinitrotoluene. After acidic hydrolysis, which served to release the conjugated part of the 2,4-dinitrobenzoic acid, the analyte was selectively separated from the urine matrix via various extraction steps and then derivatised to the methyl ester. Quantitative analysis was carried out using capillary gas chromatography and mass selective detection. 3,5-Dinitrobenzoic acid was used as an internal standard. The detection limit was 1 microg l(-1) urine. The relative standard deviations of within-series imprecision were between 5 and 6%. The relative recoveries were between 91 and 110% depending on the concentration. The analytical method developed as part of this study was used to investigate a collective consisting of 82 urine samples from persons working in the area of explosives disposal. The concentrations of 2,4-dinitrobenzoic acid determined ranged from the detection limit to 95 microg l(-1) urine. The method allowed the quantification of low-level internal exposure to dinitrotoluene. PMID- 9746247 TI - Sensitive determination of methomyl in blood using gas chromatography-mass spectrometry as its oxime tert.-butyldimethylsilyl derivative. AB - A sensitive, selective and reliable method was developed to determine methomyl ?methyl-N-[(methylcarbamoyl)oxy]-thioacetimidate?, a carbamate insecticide in human blood, using gas chromatography-mass spectrometry. Dimethylglyoxime served as an internal standard (I.S.). Methomyl in the blood was converted to its oxime form by sodium hydroxide. The solution made acidic with hydrochloric acid was poured into a column packed with Extrelut. Methomyloxime and I.S. were eluted from the column with a mixture of dichloromethane-ethyl acetate-chloroform (65:25:10), transformed to tert.-butyldimethylsilyl derivatives, and analyzed by gas chromatography-mass spectrometry in the electron impact mode. The calibration curves were linear in the concentration range from 1 ng/g to 100 ng/g and 100 ng/g to at least 5000 ng/g. The lower limit of detection was 0.5 ng/g. The absolute recoveries were 72-93% and within-day coefficients of variation were 3.1 5.6% at blood concentrations of 10 and 1000 ng/g. Two practical forensic applications are described. PMID- 9746248 TI - Determination of albumin adducts of (+)-anti-benzo[a]pyrene-diol-epoxide using an high-performance liquid chromatographic column switching technique for sample preparation and gas chromatography-mass spectrometry for the final detection. AB - A novel method has been developed for the determination of (+)-anti benzo[a]pyrene-diol-epoxide [(+)-anti-BPDE] albumin adducts in the low-picogram range. Blood from rats and humans was investigated for the validation of the method. Instead of the usual acid hydrolysis we used alkaline conditions for the cleavage of the esters formed with asparagic or glutamic acid residues of albumin. Alkaline hydrolysis gave rise to benzo[a]pyrene-r-7,t-8,t-9,c-10 tetrahydrotetrol (BT I-1) which was separated from the matrix by HPLC with a column switching technique. The analytes were collected by an automated fraction collector and after silylation determined with GC-MS using negative chemical ionization. Adduct concentrations were calculated by the internal standard method. Benzo[a]pyrene-r-7,t-8,c-9,c-10-tetrahydrotetrol (BT-II-2) was used as an internal standard because of its similar physicochemical properties and its absence from human samples. To determine the recovery of the analytical procedure benzo[a]pyrene-r-7,t-8,t-9,t-10-tetrahydrotetrol (BT I-2) was added at the end of the sample clean-up. Single ion recording mode was applied for the detection of the analyte and the standards using the abundant fragment ion m/z 284 for quantitation of the three tetrols. The mean recovery of the internal standard BT II-2 was about 50%. The limit of detection was 0.15 pg per injection corresponding to 0.01 fmol/mg albumin. Regression coefficients of the calibration curves were r2=0.99 and r2=0.98 for BT I-1 concentration ranges of 4-400 ng/l and 4-40 ng/l, respectively. The mean coefficient of variation for duplicate analyses of human albumin samples was found to be 22%. PMID- 9746249 TI - Development of a coupled-column liquid chromatographic-tandem mass spectrometric method for the direct determination of betamethasone in urine. AB - Different hyphenated liquid chromatographic (LC) and mass spectrometric (MS) techniques were investigated in order to set-up a method for the fast, direct analysis of betamethasone in hydrolysed and non-hydrolysed urine using large volume sample injection. After the optimisation of the LC parameters using a traditional UV detector and of the thermospray and mass spectrometric parameters by flow injection, urine samples (0.5 ml) were submitted to analysis by either LC combined with tandem mass spectrometry (MS-MS), coupled-column LC (LC-LC) combined with single quadrupole MS, and LC-LC-MS-MS. Both the three-step configurations (LC-MS-MS and LC-LC-MS) did not provide satisfactory results: loss of sensitivity was noted in the case of LC-MS-MS (likely due to reduced efficiency in the ionisation of betamethasone in the thermospray owing to the presence of large amounts of matrix interference), while in the case of LC-LC-MS a high chemical noise resulting in insufficient selectivity of detection was observed. On the contrary, LC-LC-MS-MS analysis proved to meet the demand of high speed of analysis (sample throughput, 4.5 h(-1)), selectivity, and sensitivity (LOQ, 1 ng/ml; LOD, 0.2 ng/ml). Notwithstanding the complex analytical system adopted, the developed procedure was manageable and very robust, provided that at the beginning of each analytical session the performance of the system was controlled by checking the retention time of the analytes on the first analytical column with UV detection and by optimising vaporiser temperature of the thermospray by flow injection. PMID- 9746250 TI - Simultaneous determination of cyanide and thiocyanate in blood by ion chromatography with fluorescence and ultraviolet detection. AB - An ion chromatographic method for the simultaneous determination of cyanide and thiocyanate in blood has been developed. After extraction by adding water and methanol to blood, cyanide was derivatized with 2,3-naphthalenedialdehyde and taurine to give a fluorescent product of 1-cyanobenz[f]isoindole. This compound was detected with high sensitivity by fluorometry and the underivatized thiocyanate was detected by ultraviolet absorption. The detection limits were 3.8 pmol ml(-1) for cyanide and 86 pmol ml(-1) for thiocyanate, and the recoveries from blood were ca. 83% and ca. 100%, respectively. The proposed method was successfully applied to the analysis of both anions in blood from smokers, non smokers and fire victims. PMID- 9746251 TI - Determination of flunitrazepam and its metabolites in blood by high-performance liquid chromatography-atmospheric pressure chemical ionization mass spectrometry. AB - A selective assay of flunitrazepam (F) and its metabolites 7-aminoflunitrazepam (7-AF), N-desmethylflunitrazepam (N-DF) and 3-hydroxyflunitrazepam (3-OHF) with liquid chromatography-atmospheric pressure chemical ionization mass spectrometry (LC-APCI-MS, positive ions) is described. The drugs were isolated from serum, blood or urine using a solid-phase extraction procedure previously applied to various drugs of abuse. F-d3 and 7-AF-d3 were used as internal standards. The drugs were separated on ODS column in acetonitrile-50 mM ammonium formate buffer, pH 3.0 (45:55, v/v). After analysis of mass spectra taken in full scan mode, a selected-ion monitoring detection was applied with following ions: m/z 284 (7-AF and F), 287 (7-AF-d3 and F-d3), 314 (F), 300 (N-DF and 3-OHF), 317 (F-d3), 330 (3 OHF). The limits of detection were: 0.2 microg/l for F and 7-AF, 1 microg/l for N DF and 3-OHF. The method was linear in the range 1-500 microg/l, the recoveries ranged from 92 to 99%. The method was applied for determination of F and metabolites in clinical and forensic samples. LC-APCI-MS seems to be a method of choice for these compounds. PMID- 9746252 TI - Determination of a novel selective inhibitor of type 1 5alpha-reductase in human plasma by liquid chromatography with atmospheric pressure chemical ionization tandem mass spectrometry. AB - A sensitive and specific assay of human plasma for the determination of (5alpha,7beta,16beta)-16[(4-chlorophenyl)oxy]-4,7- dimethyl-4-aza-androstan-3-one (I), a selective inhibitor of human type 1 5alpha-reductase, has been developed. The method is based on high-performance liquid chromatography (HPLC) with tandem mass spectrometric (MS-MS) detection. The analyte (I) and internal standard, Proscar (II), were isolated from the basified biological matrix using a liquid liquid extraction with methyl-tert.-butyl ether (MTBE). The organic extract was evaporated to dryness, the residue was reconstituted in mobile phase and injected into the HPLC system. The MS-MS detection was performed on a PE Sciex API III Plus tandem mass spectrometer using a heated nebulizer interface. Multiple reaction monitoring using the precursor-->product ion combinations of m/z 430- >114 and 373-->305 was used to quantify I and internal standard (II), respectively. The assay was validated in the concentration range of 0.5 to 500 ng/ml in human plasma. The precision of the assay, expressed as coefficient of variation (C.V.), was less than 7% over the entire concentration range, with adequate assay specificity and accuracy. The HPLC-MS-MS method provided sufficient sensitivity to completely map the 24 h pharmacokinetic time-course following a single 0.5 mg dose of I. PMID- 9746253 TI - Column-switching high-performance liquid chromatographic assay for determination of apigenin and acacetin in human urine with ultraviolet absorbance detection. AB - A high-performance liquid chromatographic (HPLC) method is described for the determination of apigenin and the 4'-methylated derivative acacetin in human urine using column-switching and ultraviolet (UV) absorbance detection. Urine samples were enzymatically hydrolysed and solid-phase extracted prior to injection onto the HPLC system. Prior to elution of apigenin and the internal standard, 5,7,8-trihydroxyflavone, from the first column used for sample clean up, the six-port valve was switched to the second column for analysis with UV detection. Detection of apigenin was precise and reproducible, with a limit of quantification of 10 ng ml(-1) urine. Detection and quantification of acacetin was linear down to 70 ng ml(-1) urine. The method has been successfully applied to determine the level of apigenin in 100 human urine samples from an intervention study with parsley. PMID- 9746254 TI - Quantitative determination of (-)-2'-deoxy-3'-thiacytidine (lamivudine) in human plasma, saliva and cerebrospinal fluid by high-performance liquid chromatography with ultraviolet detection. AB - A high-performance liquid chromatographic method for the quantitative determination of the HIV reverse transcriptase inhibitor lamivudine ((-)-2'-deoxy 3'-thiacytidine, 3TC, Epivir) in human plasma, saliva and cerebrospinal fluid is described. Lamivudine was extracted from samples using silica extraction columns prior to reversed-phase high-performance liquid chromatography with ultraviolet detection at 270 nm. The method has been validated over the range of 10 (lower limit of quantitation) to 5000 ng/ml using a 0.5-ml sample volume. Between-day and within-day precisions ranged from 3.5 to 9.0%. The assay has been used for the quantitative analysis of lamivudine in plasma and cerebrospinal fluid of HIV 1 infected patients. PMID- 9746255 TI - Rapid determination of nevirapine in human plasma by ion-pair reversed-phase high performance liquid chromatography with ultraviolet detection. AB - Nevirapine is a non-nucleoside reverse transcriptase inhibitor for the treatment of HIV-1-infected patients. A simple and rapid high-performance liquid chromatographic method for the quantification of nevirapine in human plasma is described. Sample pretreatment consists of protein precipitation with acetonitrile. The analyte is separated from endogenous compounds by isocratic reversed-phase, ion-pair, high-performance liquid chromatography with ultraviolet detection at 282 nm. The method has been validated over the range of 52-10400 ng/ml using 250 microl of plasma. The assay was linear over this concentration range. Within- and between-day precisions were less than 4.5% for all quality control samples. The lower limit of quantitation was 52 ng/ml. Recovery of nevirapine from human plasma was 94.5%. This validated assay is suited for use in pharmacokinetic studies with nevirapine and can readily be used in a hospital laboratory for the monitoring of nevirapine concentrations. PMID- 9746256 TI - Size-exclusion chromatographic study of the reduction of recombinant hepatitis B surface antigen. AB - The reduction of the P. pastoris-derived hepatitis B surface antigen (HBsAg) has been investigated by size exclusion chromatography performed in a detergent solution containing 0.3% sodium dodecyl sulfate (SDS) and 0.1 M Tris-HCl, pH 7.0. The HBsAg, reduced under different conditions and passed through the TSK G4000 SW column (600x7.5 mm I.D.) at 0.9 ml min(-1), was resolved into two peaks corresponding to the reduced, monomeric, and non-reduced forms, respectively. Under these conditions, the antigen fraction corresponding to the HBsAg dimer can be separated and completely reduced to monomers by repeated reductive treatment with simultaneous lipid removal. The efficiency of reduction was maximal after sample treatment with an equal volume of a solution containing 417 mM dithiothreitol, 4.2% (w/v) SDS and 16% (v/v) 2-mercaptoethanol. In conclusion, complete reduction of recombinant HBsAg to monomer subunits is possible and depends on the efficiency of lipid removal during the reductive treatment. PMID- 9746257 TI - Determination of montelukast sodium in human plasma by column-switching high performance liquid chromatography with fluorescence detection. AB - MK-0476 (montelukast sodium) is a potent and selective cysteinyl leukotriene receptor antagonist that is being investigated in the treatment of asthma. A simple and sensitive method for the determination of MK-0476 in human plasma was developed using column-switching high-performance liquid chromatography (HPLC) with fluorescence detection. A plasma sample was injected directly onto the HPLC system consisting of a pre-column (Capcell pak MF) and an analytical column (Capcell pak C18) which were connected with a six-port switching valve. The column eluate was monitored with a fluorescence detector (excitation at 350 nm; emission at 400 nm). The calibration curve was linear in a concentration range of 1-500 ng ml(-1) for MK-0476 in human plasma. The intra-day coefficients of variation of all concentrations within the range was less than 9.2%, and the intra-day accuracy values were between 97.2 and 114.6%. This method was used to measure the plasma concentration of MK-0476 following oral administration of the drug in humans. PMID- 9746258 TI - High-performance liquid chromatographic determination of the magnetic resonance imaging contrast agent gadobenate ion in plasma, urine, faeces, bile and tissues. AB - The gadobenate ion is an intravascular paramagnetic contrast agent for magnetic resonance imaging. An HPLC method for assaying gadobenate ion in plasma, urine, faeces, bile and tissue samples is described. The analysis is based on the reversed-phase chromatographic separation of gadobenate ion from the endogenous components of biological matrices and detection by UV absorption at 210 nm. The selectivity of the method was satisfactory. The mean absolute recovery was greater than 95%. The precision and accuracy of the analytical methods were in the range 0.1-6.5% and -12 to +9.3%, respectively. The detection limits in plasma (0.1 ml), urine (0.05 ml), dried faeces (200 mg suspended in 4 ml water), bile (0.5 ml), and dried liver tissue (100 mg suspended in 1 ml water) were, respectively, 0.24, 0.47, 2.6, 0.63 and 2.8 nmol ml(-1) (corresponding to 0.16, 0.31, 1.7, 0.42 and 1.9 microg ml(-1)). PMID- 9746259 TI - Urinary organic acid screening by solid-phase microextraction of the methyl esters. AB - We developed a new sample preparation method for profiling organic acids in urine by GC or GC-MS. The method includes derivatisation of the organic acids directly in the aqueous urine using trimethyloxonium tetrafluoroborate as a methylating agent, extraction of the organic acid methyl esters from the urine by solid-phase microextraction, using a polyacrylate fiber with a thickness of 85 microm and transfer of the methyl esters into the GC or the GC-MS instrument. Desorption of the analytes takes place in the heated injection port. The proposed sample preparation is very simple. There is no need for any evaporation step and for the use of an organic solvent. The risk of contamination and the loss of analytes are minimized. The total sample preparation time prior to GC or GC-MS analysis is about 40 min, and therefore more rapid than other sample preparation procedures. The urinary organic acids are well separated by GC and 29 substances are identified by GC-MS. PMID- 9746260 TI - Improvement in the high-performance liquid chromatography malondialdehyde level determination in normal human plasma. AB - We report a very rapid and simple isocratic reversed-phase HPLC separation of malondialdehyde (MDA) in normal human plasma without previous purification of the MDA-2-thiobarbituric acid (TBA) complex. The separation of MDA-TBA complex was performed using a 250x4.6 mm Nucleosil-5C18 column with a mobile phase composed of 35% methanol and 65% 50 mM sodium phosphate buffer, pH 7.0. Samples of 50 microl (composed of 100 microl plasma mixed with 1.0 ml of 0.2% 2-thiobarbituric acid in 2 M sodium acetate buffer containing 1 mM diethylenetriaminepentaacetic acid, pH 3.5, and 10 microl of 5% 2,6-di-tert.-butyl-4-methylphenol in 96% ethanol, incubated at 95 degrees C for 45 min [K. Fukunaga, K. Takama and T. Suzuki, Anal. Biochem., 230 (1995) 20] were injected into the column. The MDA-TBA complex was eluted at a flow-rate of 1 ml/min and monitored by fluorescence detection with excitation at 515 nm and emission at 553 nm. Analysis of groups of normal male and female volunteers gave plasma levels of MDA of 1.076 nmol/ml with a coefficient of variation of about 58%. No significant statistical differences were found between male and female groups, and no correlation was discovered on the age. PMID- 9746261 TI - New method for determination of fecal sterols in urine using non-chlorinated solvents. AB - A new method has been developed to determine a number of sterols in urine using non-chlorinated solvents, namely methyl tert.-butyl ether and methanol (the MTB method). The method involves liquid-liquid extraction, saponification, reextraction, silylation and final identification and quantification by GC-FID. The sterols determined were coprostanol, epicoprostanol, cholesterol and dihydrocholesterol. 5Alpha-cholestane was used as internal standard. The limit of detection for the sterols in this experiment was 2 microg l(-1) urine. Recovery of coprostanol over the range 5-100 microg l(-1) urine by this method was between 80 and 92%. PMID- 9746262 TI - Stereoselective determination of clenbuterol in human urine by capillary electrophoresis. AB - The effectiveness of capillary electrophoresis (CE) in the field of stereoselective determination of drugs in biological matrices is demonstrated by analyzing clenbuterol in human urine. Due to the very low therapeutical doses of 20-40 microg per day the total concentrations in urine are 1-10 ng/ml. The sample was extracted with hexane-tert.-butyl methyl ether (99.5:0.5). The reconstituted sample was injected electrokinetically (50 s, 10 kV). Using phosphate buffer, pH 3.3 and hydroxyethyl-beta-cyclodextrin as chiral selector the total analysis time was below 15 min. The limit of determination was estimated as 0.5 ng/ml per enantiomer. S-(-)-Bupranolol was used as internal standard. Both precision and accuracy of the method were within the limits for biological samples. The application to human urine from patients having received therapeutical doses showed a slightly predominant excretion of the (+)-enantiomer to the (-) enantiomer. PMID- 9746263 TI - Development of a sensitive liquid chromatography-electrospray ionization tandem mass spectrometry method for the measurement of 7-cyanoquinocarcinol in human plasma. AB - A sensitive method for the determination of an anti-cancer agent, DX-52-1 (7 cyanoquinocarcinol, I) and quinocarmycin (II) which is formed from I either by metabolism or degradation, in human plasma has been developed utilising liquid chromatography electrospray-ionization tandem mass spectrometry (LC-ESI-MS-MS). The procedure involves solid-phase extraction at pH 2 and low temperature (4-6 degrees C) to prevent the decomposition of I to II, the separation by reversed phase HPLC and the multiple reaction monitoring (MRM) by ESI-MS-MS. The mean precision and accuracy at the lower limit of quantitation (LLOQ) of I, 0.25 ng ml(-1), were 8.7% and - 10.8%, respectively. Since an interfering peak eluting slightly earlier than II was observed on the HPLC of blank plasma, the LLOQ of II was set at 5 ng ml(-1) where the mean precision and accuracy were 15.6% and 9.8%. The results suggested that the method is useful for the simultaneous monitoring of I and II in the clinical trials of I. PMID- 9746264 TI - Rapid simple high-performance liquid chromatographic determination of paroxetine in human plasma. AB - A rapid, simple method for the measurement of paroxetine in human plasma by reversed-phase high-performance liquid chromatography (HPLC) with fluorescence detection is described. This method includes only one-step extraction of paroxetine and dibucaine, an internal standard, with chloroform. Their recoveries were around 90%. The mobile phase, 10 mM phosphate buffer-acetonitrile (40:60, v/v) was eluted isocratically. Between- and within-day coefficients of variation were in the range of 1.9-9.4% and 2.3-13.3%, respectively. The detection limit was 0.2 ng/ml. The method we describe can be easily applied to the measurement of plasma paroxetine concentration for pharmacokinetic studies as well as for therapeutic drug monitoring in patients taking paroxetine. PMID- 9746265 TI - Folate deficiencies and cardiovascular pathologies. AB - Although folates are widely distributed in foods, folate deficiencies may be more frequent than expected because their true availability may be impaired due to their lability under various food cooking and processing conditions. Folate deficiency is frequently observed in elderly people, smokers, alcoholics and oral contraceptive users. It is also associated with the mutation leading to the thermolabile variant of N5,10-methylenetetrahydrofolate reductase which is observed in about 10% of the population. In addition to the essential role of the intracellular pool of polyglutamates in de novo biosynthesis of deoxyribonucleotides which allow cell growth and division, the reduced and methylated form of folate, N5-methyltetrahydrofolate, is required for the remethylation of homocysteine to methionine. By inhibiting this remethylation pathway, folate deficiency induces homocysteine efflux into the circulation. Many studies have shown a negative correlation between plasma folate, particularly N5 methyltetrahydrofolate, and circulating homocysteine levels. In addition, folate deficiency is a major cause of hyperhomocysteinemia which is fully recognised as an independent risk factor for atherothrombosis. Epidemiological and recent experimental studies have demonstrated that folate deficiency might increase the risk of cardiovascular disease by increasing circulating homocysteine levels. Thus, the clinical efficiency of folate supplementation, especially N5 methyltetrahydrofolate, in reducing homocysteine-dependent cardiovascular risk should be evaluated. PMID- 9746266 TI - Paraoxonase as a risk marker for cardiovascular disease: facts and hypotheses. AB - Paraoxonase (PON1) is a Ca2+-dependent enzyme whose mechanism of action is incompletely elucidated. PON1 was originally found to be responsible for the hydrolysis of paraoxon, a catabolite of the insecticide parathion, but this enzyme is equally able to hydrolyze other substrates such as phenyl acetate. PON1 exhibits two sequence polymorphisms, Arg-->Gln 192 and Met-->Leu 55, respectively, of which the former is responsible for the distinct catalytic activity of the two corresponding allozymes against paraoxon. The PON1 gene is a member of a family of at least three related genes. Although the physiologic substrate of PON1 is unknown, a protective role against the oxidative degradation of serum lipoproteins has been attributed to this enzyme. Indeed, PON1 is a component of a spectrum of circulating high density lipoprotein particles and can hydrolyze oxidized phospholipids and cholesteryl ester hydroperoxides. Studies have been conducted to evaluate the possible "protective" role of PON, and especially the influence of the Arg-->Gln 192 polymorphism, in coronary artery disease. Results from these investigations are conflicting, and recent data suggest a complex pattern with influences from other polymorphisms in either the PON1 and/or the PON2 and PON3 genes, or even another region of the gene cluster. A number of related factors, which include the heterogeneity of the high density lipoprotein particles incorporating PON(s), the metabolism of associated apolipoproteins such as apoJ/clusterin, the respective roles of PON(s) and other high density lipoprotein-associated enzymes such as platelet-activating-factor acetyl-hydrolase and lecithin-cholesterol acyltransferase, modifications of high density lipoprotein composition and activity under acute-phase conditions, the dietary and environmental regulation of PON(s), and the actual in situ availability of PON in the atherosclerotic artery wall, must equally be taken into account. PMID- 9746267 TI - Molecular diagnosis of lecithin: cholesterol acyltransferase deficiency in a presymptomatic proband. AB - We report the molecular diagnosis of a lecithin : cholesterol acyltransferase deficiency in a 12-year old proband with a high-density lipoprotein deficiency. The increased percentage of free cholesterol in plasma and high-density lipoprotein indicated an inherited lecithin : cholesterol acyltransferase deficiency as the underlying cause. This diagnosis was confirmed by a low plasma lecithin : cholesterol acyltransferase activity and a combination of genetic analyses which demonstrated compound heterozygosity for two mutations in the lecithin : cholesterol acyltransferase gene of the proband. One was a previously unreported 2 bp deletion leading to a stop signal in codon 77 and the other a point mutation causing Arg 135-->Gln transition. To our knowledge, this is the first diagnosis of lecithin : cholesterol acyltransferase deficiency in a pre symptomatic patient. Whether the proband will develop signs of complete lecithin : cholesterol acyltransferase deficiency or the milder form (Fish Eye Disease) is uncertain, although the former possibility is more likely. The risk of premature atherosclerosis conferred by lecithin : cholesterol acyltransferase deficiency is not well established. The proband will need to be carefully monitored in the future. PMID- 9746268 TI - Cathepsin D and cathepsin L activities in aortic aneurysm wall and parietal thrombus. AB - Deterioration of the aortic wall resulting in formation of aneurysms may be caused by increased activity of metalloproteases and lysosomal proteases. The aim of this work was the evaluation of cathepsin D and cathepsin L activities, and activities of inhibitors of cysteine cathepsins in the wall of aortic aneurysms and in parietal thrombus. Aortic aneurysms were obtained during operation. Aortas taken from organ donors and blood clots were used as control material. Activities of cathepsin D and cathepsin L in the aortic aneurysm wall and parietal thrombus were higher than in the control groups. The aneurysm wall showed lower activity of inhibitors of cysteine proteases than the normal aorta. Parietal thrombus had a higher level of cysteine protease inhibitor activity than blood clot. Cathepsin D and cathepsin L present in the aneurysm wall and in the parietal thrombus filling the aneurysm may act on proteins determining elasticity and mechanical resistance of arteries. PMID- 9746269 TI - The diagnostic value of serum homocysteine concentration as a risk factor for coronary artery disease. AB - Hyperhomocysteinemia is now regarded as an established risk factor for coronary artery disease and is present frequently in the general population. However, the diagnostic value of this risk factor relative to others has only occasionally been investigated. We compared the diagnostic value of classic risk factors and of homocysteine in a retrospective case-control study in 191 cases with angiographically established coronary artery disease and 231 healthy controls. Life style habits were assessed by a detailed questionnaire. Laboratory parameters including lipoproteins and blood lipids, homocysteine, folate, and vitamin B12 were measured and their diagnostic value compared with each other by use of receiver-operator characteristic analysis. Comparison of the receiver operator characteristic curves revealed that homocysteine significantly discriminated between cases and control subjects. High-density-lipoprotein cholesterol, triglycerides and non-esterified fatty acids also had an area under the curve significantly different from 0.5 (the area under the curve representing no discrimination). Homocysteine was weakly related to folate, vitamin B12, age and serum creatinine concentration. We conclude that hyperhomocysteinemia is at least as important as conventional risk factors for coronary artery disease and that receiver operator characteristic analysis of homocysteine is suitable to determine patients at the highest risk for coronary artery disease. Clinical trials testing the effect of homocysteine lowering by vitamin supplementation in the prevention of coronary artery disease are needed. PMID- 9746270 TI - Influence of time and temperature on coagulation analytes in stored plasma. AB - There is no comprehensive study on the stability of coagulation analytes in plasma. We therefore determined the influence of storage time and temperature on prothrombin time, activated partial thromboplastin time, thrombin time, fibrinogen, factors V and VIII, antithrombin III, protein C and S in plasma from 20 healthy subjects and 20 patients receiving heparin therapy. The stability in plasma, defined as the period during which there was a change of less than 10% from the initial value, was 8 hours for activated partial thromboplastin time, 24 hours for prothrombin time, 48 hours for factor V and 7 days for thrombin time, fibrinogen, protein C and antithrombin III in healthy subjects at 6 degrees C. Factor VIII and protein S showed 19 and 12 % reduction in activity, respectively, after 8 hours. In volunteers not treated with heparin therapy, activated partial thromboplastin time was stable for 8 hours; prothrombin time for 48 hours; and thrombin time, fibrinogen and antithrombin III for 7 days with sample storage at room temperature. Factor VIII showed a decrease of 18 % after 8 hours. For patients receiving heparin therapy, the stability of the analytes in plasma stored at 6 degrees C was 8 hours for thrombin time, 24 hours for prothrombin time and activated partial thromboplastin time and 7 days for fibrinogen and antithrombin III. Factors V and VIII showed a decrease of 13 % and 20 % respectively after 8 hours. When the plasma of these patients was stored at room temperature, factor V was stable for 8 hours, and prothrombin time for 24 hours, whereas fibrinogen and antithrombin III remained unchanged for 7 days. Activated partial thromboplastin time showed an increase of 13 %, thrombin time a fall of 16 %, and factor VIII a decrease of 18 % after 8 hours. PMID- 9746271 TI - The uncertainty associated with the predictive value of test results. AB - The uncertainty associated with predictive value of test results was taken into consideration, as concerns both sampling error (related to the size of the statistical reference samples) and analytical imprecision (unavoidably involved by the measurement itself). A software package, developed for the statistical calculations, was used for the treatment of the results obtained for serum free thyroxin in euthyroid and dysthyroid subjects, assumed as an experimental model. Examples are shown for the obtainable functions predictive value vs. estimate and the related uncertainty regions. These data could help in comparing test results and, in particular, in preparing fully informative laboratory reports. The difficulties involved by an extensive application of the procedure are discussed. PMID- 9746272 TI - Clinical evaluation of the CARDIAC STATus, a rapid immunochromatographic assay for simultaneous detection of elevated concentrations of CK-MB and myoglobin in whole blood. AB - We studied the performance of the CARDIAC STATus, a new rapid, easy to perform qualitative whole blood bedside test for detection of elevated CK-MB and myoglobin in the emergency room. Blood samples from 182 consecutive patients with chest pain were drawn on admission and at five and seven hours after the onset of symptoms. The CARDIAC STATus tests were performed by coronary care unit nurses and, independently, by a trained laboratory technician. The results were compared with quantitative assays for CK-MB mass and myoglobin. At the end of the study, a second test series using a new lot number of cartridges was performed on the same blood samples because of possible elution buffer contamination. Nurses produced more false negative results than the technician (CK-MB 43 vs. 27 %, p=0.01, myoglobin 31 vs. 13%, p<0.0001), but the technician produced more false positive myoglobin results (9.3 vs. 5.5%, p=0.0001). In the second test series, the nurses produced significantly fewer false negative tests both for CK-MB (19%, p<0.0001) and myoglobin (13%, p=0.0002). The false negative rate for the technician was not different between the first and the second test series. The CARDIAC STATus yields a substantial number of false negative results both for CK-MB and myoglobin when compared to a quantitative assay, and therefore at present has limited value for ruling out an acute myocardial infarction. PMID- 9746273 TI - Multicentre evaluation of KONE Optima analysis system. AB - Optima from KONE Instruments Corporation is a new selective laboratory analyzer for turbidimetric, colorimetric and ion selective electrode measurements. Overall analytical performance of Optima and reagents provided by KONE was evaluated according to ECCLS guidelines, in a multicentre study involving four different laboratories, including substrates (cholesterol, high-density lipoprotein cholesterol, creatinine), specific proteins (transferrin, IgG), enzyme activities (gamma-glutamyltransferase, alanine aminotransferase) and electrolytes (sodium, potassium, chloride). The results obtained attest good precision of the system for all the analytes tested: the range of within-run CV is 0.5 %-4.3 %, and range of between-day CV % is 0.8 %-7.9 % (median of four laboratories). Except for total cholesterol (5 % overestimation compared to the reference method) accuracy of measurement is adequate. Creatinine and uric acid assays were subjects of interference (defined as deviation > 10 % from target value) by bilirubin and haemoglobin respectively. PMID- 9746274 TI - Biological day-to-day variation and daytime changes of testosterone, follitropin, lutropin and oestradiol-17beta in healthy men. AB - Information on biological day-to-day variation is needed for detecting within subject changes over time. In this study the daytime changes and the biological day-to-day variation of serum testosterone, follitropin, lutropin and oestradiol 17beta concentrations were investigated in 31 healthy males. To analyse daytime changes, blood specimens were taken at 0800 h, 1200 h, 1600 h and 2000 h during one day (n=31) and two days (n=8). The day-to-day variation was analysed from blood specimens collected at 0800 h on days 1 and 2 (n=31) and additionally on days 3, 4, 6 and 9 (n=8). The evaluation of the day-to-day variation was based on calculations of the within-subject (CVA+I) and between-subject (CV(G)) coefficients of variation. When the within-subject day-to-day variances were not too heterogeneous, they were used for the calculation of 95 % reference change limits. Serum testosterone and oestradiol-17beta concentrations showed a significant daytime variation; testosterone had higher serum concentrations at 0800 and 1200 h. A peak in the serum concentration of oestradiol-17beta occurred at 1200 h with a decrease towards the evening. There were no clear daytime changes in the serum concentrations of follitropin or lutropin. For different analytes the reference change limits were: serum testosterone +/- 32.0 %, serum follitropin +/- 24.1 % and serum oestradiol-17beta +/- 38.3 %. The reference change limit was not calculated for serum lutropin, as a high degree of heterogeneity and individuality was found. The interpretation of the results of hormone measurements requires recognition of the biological daytime and day-to day changes of hormones. The reference change limits determine what changes are significant when monitoring the patient. PMID- 9746275 TI - Qualitative immunoglobulin abnormalities in HIV-positive patients: long-term follow-up. AB - We studied the immunoglobulins of a cohort of 212 HIV-positive patients followed up for 13 years. The qualitative study of immunoglobulins by immunoelectrophoresis and immunofixation distinguished three groups of patients: those with monoclonal immunoglobulins, those with minor abnormalities of immunoglobulins and those with polyclonal immunoglobulins. We characterized these groups according to age, sex, immunoglobulin isotypes, and survival curves. The results show that this population of immunoglobulinopathies has distinctive characteristics. In particular, the presence of immunoglobulin abnormalities has no significant prognostic value. PMID- 9746276 TI - Difference plots and mountain plots are useful also in comparing CA 125 immunoassay systems manufactured by the same company. PMID- 9746277 TI - Decoquinate induces tissue cyst formation by the RH strain of Toxoplasma gondii. AB - Decoquinate is an anticoccidial agent that inhibits respiration in the parasites mitochondrion. We examined human foreskin fibroblast cell cultures infected with the normally tissue cyst-less RH strain of Toxoplasma gondii and treated with decoquinate for evidence of tissue cyst induction and formation. Transmission electron microscopy observations demonstrated tissue cysts in decoquinate-treated cultures on days 3, 4, 5, and 6 after inoculation. Tissue cysts contained a tissue cyst wall that enclosed stages that resembled tachyzoites and stages that were structurally bradyzoites. Similar treatment of human foreskin fibroblast cells infected with tachyzoites of the TS-4 temperature-sensitive mutant of the RH strain did not result in production of tissue cysts. PMID- 9746278 TI - Isolation of Theileria taurotragi and Theileria mutans from cattle in Botswana. AB - Two Theileria species demonstrated in peripheral Giemsa-stained blood smears of sick cattle from various parts of Botswana were subsequently identified as Theileria mutans and T. taurotragi using DNA hybridization and the polymerase chain reaction. Initial screening for Theileria species was done using microscopy, the indirect fluorescent antibody technique and the micro Elisa test. The syndrome was characteristically that of high morbidity but low mortality and vague malaise. A low parasitaemia of pleomorphic intra-erythrocytic piroplasms and the absence of schizont stages in circulating lymphocytes and lymph node aspirates were evident. Dual infections were common and often complicated by intercurrent disease conditions. PMID- 9746279 TI - Apparent digestibility of nutrients and nitrogen balance during experimental infection of calves with Eimeria bovis. AB - The apparent digestibilities (AD) of dry matter (DM), organic matter (OM), crude ash (CA), crude fiber (CFi), crude fat (CFa) crude protein (CP) and nitrogen-free extracts (NFE) and the nitrogen balance were investigated during experimental Eimeria bovis coccidiosis in calves. noninfected pair-fed controls and controls fed on a normal plan of nutrition were included in the study to allow differentiation between the effects of infection and of changes in feed intake. Primary infection with 5 x 10(4) oocysts (n = 4, group A) caused mild diarrhea and calves infected primarily with 1 X 10(5) oocysts (n = 5, group B) suffered from mild (three calves) to severe hemorrhagic (two calves) diarrhea. No clinical disease was seen after reinfection of the group A calves with 1 X 10(5) oocysts. The primary infection with 5 X 10(4) oocysts or reinfection with twice the primary inoculum did not affect AD of nutrients or the overall nitrogen balance (RT). AD of DM, NFE or OM were higher in group B during patency and in the pair fed group C calves (n = 5) than in the reinfected but healthy group A calves. AD of CFi of the group B calves even exceeded the values of the pair-fed controls. The two calves of group B that suffered from hemorrhagic diarrhea and anorexia had low values of AD of CP during the acute phase of the disease and the plasma nitrogen levels were reduced in this group. Severe clinical coccidiosis transiently reduced the nitrogen balance. It is concluded that the transient increase of AD of nutrients, especially of CFi, during clinical coccidiosis reflect hypomotility and that anorexia and intestinal leakage impair the nitrogen balance and cause weight depression. PMID- 9746281 TI - Purification of Trichinella spiralis tubulin: comparison of several analytic procedures. AB - This study compares the purity indices found after purifying tubulin obtained from the nematode parasite Trichinella spiralis, using different chromatographic and electrophoretic methods. Affinity chromatography, using monoclonal antibodies anti-alpha and anti-beta-tubulin fixed to activated Sepharosa 4B-CNBr, yields a tubulin purity of 15%. In contrast, by means of interchange-anionic chromatography using a column of DEAE-Sephadex-A50, we obtained an increase in purity of up to 75%. Finally, with the combined application of preparative electrophoresis and electroelution of proteins in polyacrylamide gels with SDS, we obtained the best results with a purity reaching 90%. PMID- 9746280 TI - Use of excretory/secretory antigens in a competition test to follow the kinetics of infection by Fasciola hepatica in cattle. AB - Eight 16-18-month-old Charolais heifers were experimentally infected with Fasciola hepatica. An antigen competition assay was used to follow the kinetics of the infection and was compared to antibody tires and serum liver enzymes. The antigen competition assay was able to detect the presence of infection as soon as 6 days after the start of the experimental infection which is considerably sooner than other methods. Consequently, this assay would be useful in diagnosing fasciolosis early in the prepatent period. The animals were slaughtered at the end of the experiment, the livers recovered and post-mortem fluke burdens determined. However, only serum liver enzyme levels gave any indication of the intensity of infection in the different animals. PMID- 9746282 TI - Breed and season effects on the peri-parturient rise in nematode egg output in indigenous ewes in a cool tropical environment. AB - A study was carried out at the International Livestock Research Institute (ILRI) Debre Berhan Research Station in Ethiopia from 1992 to 1995 to compare the peri parturient rise (PPR) in faecal nematode egg counts (FEC) in ewes of two indigenous sheep breeds. A total of 1439 Menz and 1347 Horro ewes were single sire mated following oestrus synchronization to lamb in the wet and dry season. Three ewe treatment groups were constituted as mated/lactating/undrenched; mated/lactating/drenched; unmated/undrenched for three wet and three dry lambing seasons. All ewes grazed naturally contaminated pasture. Levels of faecal egg output were monitored at mating, 3 months after mating, 2 weeks before lambing, 4, 8 and 12 weeks post-lambing. A significant PPR in FEC occurred 2 weeks before lambing and peaked at 4 weeks post-parturition in ewes lambing just before the beginning of the dry season (October/November). There was no significant increase in FEC when lambing occurred before the onset of the long rainy season (May/June). The PPR in this study was associated with both lactation and seasonal availability of third-stage infective larvae on pasture. There was no consistent breed difference in FEC during the six sampling periods from mating to weaning. Faecal cultures and worm counts from both breeds confirmed the presence of Longistrongylus (Pseudomarshallagia) elongata, Trichostrongylus spp.and Haemonchus contortus. The role of the peri-parturient rise of FEC in ewes in gastrointestinal nematode transmission is discussed. PMID- 9746283 TI - The relationship between faecal egg count reduction and the lethal dose 50% in the egg hatch assay and larval development assay. AB - The relationship between resistance detected in the faecal egg count reduction test (FECRT) and the lethal dose 50% (LD50) in the egg hatch assay (EHA) for benzimidazoles (BZs) and a larval development assay (LDA) for BZs, levamisole (LEV) and ivermectin (IVM) was examined on 13 sheep farms and 12 goat farms in Denmark. Out of 10 farms where resistance to BZs was detected according to the FECRT, nine (90%) had LD50 values above 0.5 microM thiabendazole (TBZ) (0.1 microg TBZ/ml) in the EHA, indicating resistance to BZs. However, four out of the 12 isolates susceptible to BZs in the FECRT had LD50 values higher than 0.5 microM TBZ in the EHA. For all isolates examined, LD50 values for TBZ in the LDA were lower than in the EHA. Four out of 11 and five out of 12 farms with worm populations resistant to BZs according to the FECRT and EHA respectively, had LD50 values lower than 0.5 microM TBZ in the LDA. Using the same cut-off point for resistant isolates in the LDA as in the EHA (0.5 microM TBZ), these isolates would be considered susceptible to BZs. All 10 isolates susceptible to BZs according to the FECRT and EHA and two isolates with suspect BZ resistance had LD50 values lower than 0.5 microM TBZ in the LDA. The above results indicated fairly good agreement in the detection of BZ resistance between the FECRT, EHA and the LDA. Groups of farms where resistance to LEV was detected according to the FECRT had higher mean LD50 values compared to those with LEV-susceptible or suspected resistant isolates. However, only four out of 12 farms having isolates resistant to LEV had LD50 values higher than 1.2 microM LEV (0.28 microg LEV/ml) recorded previously for a LEV-susceptible strain of Ostertagia circumcincta. This indicated discrepancies in declaring resistance to LEV between the FECRT and the LDA. Isolates from four farms where resistance to IVM was detected in the FECRT had LD50 values higher than the susceptible isolates. These were 2.5 to 7.5 times higher than those recorded previously for IVM-susceptible strains. PMID- 9746284 TI - Congenital trichinellosis in the rat. AB - 57 female rats were divided into four groups according to age and gestation. Trichinella spiralis infection was induced by feeding the rats with rat muscle containing about 10000 larvae per gram. The mating and sacrificing of females were done differently for each group. New-born rats were examined by direct trichinoscopy and by peptic digestion of muscle. We found that larvae of T. spiralis which entered the uterus of the pregnant females can pass to the foetus. It was impossible to specify the time when larvae crossed the placenta, but this crossing is made only by those larvae which did not settle and encyst in the striated muscles. The mobilisation of the encysted larvae from skeletal muscles was not demonstrated. PMID- 9746285 TI - Protostrongylid nematode infection of chamois (Rupicapra rupicapra) at the Bauges massif (French Alps). AB - From 1979 to 1985, protostrongylid infections (small lungworms) of chamois were studied at the Bauges hunting reserve, in the western Alps. Fifty-two chamois were necropsied on 24 sites and lung nematodes identified to species. From June to November 1985, a more detailed study was undertaken on four sites where chamois were the only ruminants. First-stage larvae excreted in feces and infection of snails intermediate hosts were monitored. The protostrongylid species were, arranged in decreasing order of prevalence, Neostrongylus linearis, Protostrongylus rupicaprae, Muellerius capillaris, and Muellerius tenuispiculatus. Intensity was higher in females than in males (in chamois aged 4 to 11 months compared with younger or older ones) in winter than in the other seasons. Intensity of infection increased as chamois were located at lower altitudes. The snail intermediate hosts were, in decreasing order of importance, Helix pomatia, Cepaea sylvatica and Arianta arbustorum. PMID- 9746286 TI - Helminth levels of working donkeys kept under different management systems in the Moretele 1 district of the North-West Province, South Africa. AB - In Southern Africa, where 150,000 working donkeys provide an important alternative to mechanisation in resource-poor communities, very little is known about their helminth status, or about the impact of helminths on their work output. The aim of this study was to investigate the helminth status of working donkeys under different management systems. Donkey owners in three different areas (one rural and two semi-rural) of the Moretele 1 district of North-West Province, South Africa, were visited and structured interviews were used to assess the management systems under which the donkeys were kept. Faecal samples were collected from 93 donkeys in the study once a month for 14 months. Faecal samples were analysed for nematode and trematode eggs and cultured to produce third-stage larvae for the identification of the nematode species. Final comparisons between management system subgroups, as well as between areas, age groups and sexes were made. Four management systems were identified. (1) The first system identified consisted of donkeys which were kept in a small yard at all times. They were fed hay but no supplementary food. (2) The second system consisted of donkeys which were allowed to roam freely around the village most of the time and rounded up and held in an enclosure when needed for work. (3) The third system was identical to the second management system except that the donkeys were given supplementary food during winter. (4) The fourth system was only found in the one area where each owner owned 10 ha of land and here the donkeys were allowed to roam freely on the owner's land and brought into enclosures prior to working. Helminth species composition and faecal egg count numbers differed between these four systems. The main difference noted was that donkeys from management system one showed significantly higher numbers of strongyle eggs and higher percentages of some of the strongyle larvae. Management system two had a higher Strongyloides mean egg count and prevalence than the other areas. Parascaris and Gastrodiscus egg counts differed between all four systems. Since the results showed differences in the number and species of helminths in donkeys kept under the four management systems, suggestions are made as to which management system would facilitate reduction of helminth parasites in the animals. Although supplementary feeding in Moretele 1 is fairly rare, it would seem that donkeys which do have access to better food resources have lower egg counts than donkeys on limited grazing. PMID- 9746287 TI - Experimental infection of dogs with the nasal mite Pneumonyssoides caninum. AB - A successful experimental transmission of the canine nasal mite, Pneumonyssoides caninum, is described. Some 11 weeks after repeated systemic ivermectin treatment, four Beagles were inoculated via the right nostril with 20 P. caninum mites of different sexes and life stages, obtained at the necropsy of an infected dog. The inoculated dogs and a matching uninoculated control were observed for clinical signs for 14 weeks and then euthanised. Vague upper respiratory signs and a transient minor increase in the number of eosinophils in peripheral blood were recorded in the inoculated dogs. At necropsy 4-12 P. caninum mites were found in the nasal cavities and sinuses of the inoculated dogs, but none in the control. In three out of the four infected dogs mites were found in both the right and left nasal cavities and sinuses of the skull. Since in no case more mites than the number used for inoculation were detected it is not clear if the mites managed to reproduce in the dogs. Inflammatory lesions were seen most consistently in the olfactory mucosa, respiratory mucosa and tonsils, and growth of opportunistic bacteria was observed in the tonsils of the infected dogs. The inflammatory lesions seen in the olfactory mucosa may explain why dogs infected with P. caninum sometimes appear to suffer from impaired scenting ability. PMID- 9746288 TI - Neospora caninum identification in an aborted bovine fetus in Spain. AB - Neospora caninum tachyzoites were identified in areas of the cerebrum with lesions of non-suppurative encephalitis in an aborted bovine fetus in Spain. PMID- 9746289 TI - Seroprevalence of anti-Toxoplasma gondii antibodies in buffaloes in Khoozestan province, Iran. AB - The sera of 385 buffaloes of different age and sex, collected from the various areas of the Khoozestan province, Iran, were tested by the IFA test, from April 1995 to February 1996. The results indicated 8.8% (34 out of 385) buffaloes had anti-Toxoplasma gondii antibodies titered 1:16 and more. The infection rate of buffaloes under the age of a year, above a year female, and male were 10.8%, 4.7%, 12.1%, and 5.3%, respectively. PMID- 9746290 TI - Cross-reaction of an anti-Cryptosporidium monoclonal antibody with sporocysts of Monocystis species. AB - The non-specific cross-reaction of a fluorescently labelled anti-Cryptosporidium monoclonal antibody was observed microscopically when testing faecal specimens from small mammals. The reactive particles were identified as sporocysts of the Gregarine family Monocystidae, and indicate that considerable care should be taken so that false positives are not recorded. PMID- 9746291 TI - Gasterophilus intestinalis infections in horses in Belgium. AB - Over a period of one year, from December 1995 to November 1996, larvae of Gasterophilus intestinalis were found in 193 horse stomachs (58%) of 330 that were examined in two Belgian slaughter houses. When August is excluded, 62% of the stomachs had bot larvae. No other Gasterophilus species were identified. The monthly prevalence ranged from 9% in August to 75% in November and December. The mean intensity of bot larvae varied from 8 in August to 29 in March, and the majority of the horses (67%) harboured less than 50 larvae. Prevalences and intensities were not affected by age, but mares were more frequently infected than stallions and geldings (P < 0.014). The mean size of the lesions increased from 1 cm2 to 17 cm2. The majority of the larvae were localised near the margo plicatus. PMID- 9746292 TI - Activity of moxidectin 1% injectable solution against first instar Hypoderma spp. in cattle and effects on antibody kinetics. AB - The activity of the moxidectin as an 1% w/v injectable solution on first instar Hypoderma spp. has been evaluated in sixteen naturally infested young cattle. The animals were selected on the basis of their serological status and allocated to two groups of eight animals. At the end of November, one group was treated with moxidectin at a dose rate of 0.2 mg/kg via the subcutaneous route and the non treated control calves injected with the vehicle. The serological status was assessed 1, 2, 4, 8 and 12 weeks post treatment and the presence of Hypoderma lumps determined every two weeks from February to June. A 100% efficacy of the injectable formulation was demonstrated. A progressive fall of the antibody levels was observed in the treated calves for one month following treatment, suggesting a progressive action of the test compound and a limited risk of hypersensitivity. PMID- 9746293 TI - Comments on the paper "Sustainable tick and tickborne disease control in livestock improvement in developing countries". PMID- 9746294 TI - Tamoxifen modulation of carboplatin cytotoxicity in a human U-138 glioma cell line. AB - Glioma cells express high protein kinase C (PKC) activity, which may represent an important therapeutic target. Tamoxifen (TAM) has moderate PKC-inhibiting activity, blocking DNA synthesis and cellular proliferation in human glioma cells at concentrations that can be achieved therapeutically. Carboplatin (CBDCA), a second-generation platinum derivative, induces intra- and interstrand DNA-protein crosslinks producing inhibition of tumor-cell growth. In the present study, the effect of TAM, CBDCA, and the combination of both was evaluated against the human established U-138 glioma cell line during the exponential growth phase (48-72 h) by means of both the Biorad protein assay (BPA) method and Trypan blue exclusion study (TBES). Both TAM and CBDCA reduced the cellular growth rate, with a median 50%-inhibiting concentration (IC50) of 12.5 microM for TAM and 350 microM for CBDCA. The U-138 glioma cell line showed a moderate response to 100 microM of CBDCA, with < or = 10% reduction of the growth rate. The association of both chemotherapeutic agents induced a 98% reduction of the IC50 dose of TAM (0.1 microM), and a 71% reduction of the IC50 dose of CBDCA (100 microM). During the combinational TAM CBDCA exposure we observed a cytotoxic effect of TAM at concentrations lower than 0.1 microM, not recognized using it as a single drug. The differences observed among the IC50 doses (TAM, CBDCA, TAM-CBDCA) and among treated and untreated matched control cells were statistically significant (P < 0.01). Our results confirm previous observations about the efficacy in vitro of TAM against human glioma cell lines and show a marked enhancement of this activity by CBDCA. PMID- 9746295 TI - Electrophysiological comparison between corticobasal degeneration and progressive supranuclear palsy. AB - Multimodal evoked potentials were recorded in four patients with corticobasal degeneration (CBD), four patients with progressive supranuclear palsy (PSP) and 15 normal control subjects. CBD and PSP patients showed significant prolongation of the N200 and P300 latencies of auditory event-related potentials compared with controls. Patients with CBD showed significant prolongation of interpeak latencies between N13 and N20 of short-latency somatosensory evoked potentials compared with the controls and patients with PSP. The present results show that the two diseases have different electrophysiologic features. PMID- 9746296 TI - Indication to surgical management of cerebellar hemorrhage. AB - Few reports have compared patients operated for cerebellar infarcts with those operated for cerebellar hemorrhage. Considering our previous paper about patients with massive cerebellar infarcts, we report on our surgical experience with five patients with cerebellar hemorrhage. The indication for operation was decreased consciousness with signs of brainstem compression. In all patients hydrocephalus was absent or mild, as opposed to patients with cerebellar infarcts. Suboccipital craniotomy with hematoma evacuation was therefore the surgical procedure of choice. The outcome was worse than in patients with cerebellar infarcts. We conclude that depressed mental state in cerebellar hemorrhage is mainly due to pressure of the cerebellum on the activating reticular system of the brainstem. The surgical approach to patients with bleeding in the cerebellum differ somewhat from that of patients with cerebellar infarcts in timing and kind of first choice procedure. PMID- 9746297 TI - Hemifacial spasm due to posterior fossa tumors: the impact of tumor location on electrophysiological findings. AB - Ephaptic transmission is one of the electrophysiological hallmarks of hemifacial spasm. It is generally accepted that in the majority of patients with idiopathic hemifacial spasm, microvascular compression of the facial nerve at the site where the nerve exits the brain stem is the underlying cause. Whether the actual site of the ephapse is at the site of the lesion or at a nuclear level due to hyperexcitability of the facial motor nucleus is still controversial. Rarely, hemifacial spasm may be due to space occupying lesions in the cerebellopontine angle or in the brain stem. We report the electrophysiological findings of four patients with hemifacial spasm due to extra-axial tumors in different locations of the posterior fossa. The location of the tumor was intrameatal in one patient, in the cerebellopontine angle in two patients and in the brain stem in another patient. Facial nerve motor neurographies including transcranial magnetic stimulation revealed abnormal findings in two patients. Selective stimulation of facial nerve branches demonstrated delayed (ephaptic) responses in all but one patient whose hemifacial spasm had disappeared after treatment with carbamazepine. The latencies of the delayed responses did not correlate with the tumor location. In sum, the site of ephaptic transmission cannot be reliably determined by latency measurements of the delayed response because of its variability which is probably caused by the different size and diameter of the axons participating in ephaptic transmission as well as by the extent of focal demyelination at the site of the lesion. A neuroradiological work up including MR imaging should be mandatory in all patients with hemifacial spasm because electrophysiological studies fail to differentiate between idiopathic and symptomatic hemifacial spasm. PMID- 9746298 TI - Cerebral hemorrhage complicating exertional heat stroke. AB - Exertional heat stroke may be complicated by mild neurological deficits, usually with complete convalescence. This may be associated with metabolic disorders inherent to hyperthermia such as a marked coagulopathy. We report a case of a previously healthy 20 year old male who died in the course of exertional heat stroke complicated by intracerebral hemorrhage. PMID- 9746299 TI - Transient migraine-like symptoms with internal carotid artery dissection. AB - We report three patients with angiographically confirmed internal carotid artery (ICA) dissection who presented with transient symptoms resembling migraine with aura. Marching impairments from one modality to another preceded recognition of the diagnosis of dissection and were not associated with clinical or radiologic evidence of cerebral infarction. We review the clinical patterns in which ICA dissection may be identified in the setting of migrainous symptoms, given the different therapeutic approaches to migraine and dissection and the non-invasive means to diagnose dissection with magnetic resonance imaging (MRI). We offer mechanisms for recurrent neurologic symptoms in patients with ICA dissection. PMID- 9746300 TI - Dementia associated with hyperphosphatemic tumoral calcinosis. AB - Hyperphosphatemic tumoral calcinosis (HTC) is a rare inherited metabolic disorder manifested by pararticular calcification and hyperphosphatemia, caused by an elevated renal phosphate reabsorption threshold and elevated serum 1,25 dihydroxyvitamin D levels. The disorder usually affects African-American subjects, but has also been described in Caucasians and Hispanics. Vascular calcifications and angioid streaks have been reported to be associated with the disorder; however, dementia has not been previously reported. Our medical center has followed two African-American siblings, with HTC for over 30 years, who have developed dementia at 56 years and 67 years of age. Neither man has been hypertensive, however, magnetic resonance imaging (MRI) scans in both cases revealed multiple periventricular infarcts, suggestive of infarcts caused by vascular calcification in the central nervous system. These two brothers with HTC suggest that periventricular infarcts with dementia may be a long term complication of this disorder. PMID- 9746301 TI - The linear naevus sebaceus syndrome. AB - The linear naevus sebaceus syndrome (LNSS) is a phakomatosis, characterized in general by a triad consisting of naevus sebaceus of Jadassohn, seizures, and mental retardation. In addition, a broad spectrum of neurological, ophthalmological, skeletal, urogenital and cardiovascular symptoms may be encountered. According to our literature review, seizures and mental retardation were reported in 67 and 61% of cases, respectively. Because ophthalmological abnormalities (59%) and involvement of other organ systems (61%) occur frequently, we advise avoidance of adhering to the classical triad for recognizing or describing LNSS. Gross structural abnormality of the cerebrum or cranium was frequently observed (72%), consisting mainly of enlargement of one lateral ventricle, hemimegalencephaly and hemimegacranium. We report a case of a male patient with the clinical features of LNSS, but without cerebral developmental abnormalities at autopsy examination. PMID- 9746302 TI - Cystic cavernous malformation of the cerebellopontine angle. AB - A 46 year old male presented with a cerebellopontine angle syndrome. CT scan and MRI revealed a cystic mass with a solid nodule in the cerebellopontine angle. At surgery, a vascular lesion was encountered which was totally excised in two stages. The histopathology was consistent with that of a cavernous malformation. Only four such cases have been reported so far in the English literature. The clinical and radiological features and the surgical management are discussed. PMID- 9746303 TI - Ossified and de novo cavernous malformations in the same patient. AB - We report the case of a 44-year-old patient with a MRI scan showing a newly developed cavernoma after two highly calcified lesions had been excised surgically. Six other cavernous malformations had been followed by MR imaging over a 2-year period. The coexistence of the two extremes of cavernous malformations in terms of lesions development--de novo and ossified lesions has not been reported previously and has implications for both the follow-up and the natural history of these malformations. The potential for developing new cavernous malformations persists and does not seem to be related to the evolutional stage of pre-existing lesions. It is suggested that these patients need to be followed up by MRI on a regular basis. PMID- 9746304 TI - Granular cell tumor of the pituitary stalk. AB - A 52-year-old woman, who had been treated for Parkinson disease for 2 years, was found to have a suprasellar granular cell tumor. The clinical and pathological features, as well as some of the nosologic problems of this rare tumor of the neurohypophysis are briefly discussed. PMID- 9746305 TI - Sparganosis of brain and spinal cord: unusual tapeworm infestation (report of two cases). AB - Among the diseases due to cerebral parasitism, those caused by sparganum mansoni, the larval form of Spirometra mansoni, are very rare. We report two cases, one involving the frontoparietal area in a 38-year-old male and presenting as a mass lesion and another in a 10-year-old girl, presenting with paraparesis due to mid thoracic compressive mass lesion. Pathological examination of the resected lesion revealed the characteristic plerocercoid larva, spargana, enclosed in acute inflammatory exudate, resembling an abscess. Postoperative recovery was good, suggesting that the best treatment for cerebral or spinal sparganosis mansoni is surgical excision. Serological tests for diagnosis were not carried out in these cases, since parasitic infection was not suspected. Although generally the role of immunodiagnosis is limited due to rarity of the condition, in endemic areas such tests may be useful in preoperative diagnosis. PMID- 9746307 TI - Continuous lumbar cerebrospinal fluid pressure monitoring in idiopathic normal pressure hydrocephalus: predictive value in the selection for shunt surgery. PMID- 9746306 TI - Gliofibroma: mixed glial and mesenchymal tumour. Report of three cases. AB - Gliofibromas are rarely encountered astrocytic tumours comprising of astrocytic and benign fibroblastic components. They commonly occur in first two decades of life. However, the exact behaviour is not fully known and their histogenesis is also still debatable. We report three cases of gliofibroma in which we studied proliferative markers (MIB-1) and p53 protein expression. In these tumours, occurrence in adult life is in contrast to that reported in the literature. Depending upon the morphology and proliferative Labelling Index we classified these tumours into low grade (benign) and high grade (malignant/anaplastic). PMID- 9746308 TI - What do we not know of cellular bioenergetics?--a general view on the state of the art. PMID- 9746309 TI - Top-down elasticity analysis and its application to energy metabolism in isolated mitochondria and intact cells. AB - This paper reviews top-down elasticity analysis, which is a subset of metabolic control analysis. Top-down elasticity analysis provides a systematic yet simple experimental method to identify all the primary sites of action of an effector in complex systems and to distinguish them from all the secondary, indirect, sites of action. In the top-down approach, the complex system (for example, a mitochondrion, cell, organ or organism) is first conceptually divided into a small number of blocks of reactions interconnected by one or more metabolic intermediates. By changing the concentration of one intermediate when all others are held constant and measuring the fluxes through each block of reactions, the overall kinetic response of each block to each intermediate can be established. The concentrations of intermediates can be changed by adding new branches to the system or by manipulating the activities of blocks of reactions whose kinetics are not under investigation. To determine how much an effector alters the overall kinetics of a block of reactions, the overall kinetic response of the block to the intermediate is remeasured in the presence of the effector. Blocks that contain significant primary sites of action will display altered kinetics; blocks that change rate only because of secondary alterations in the concentrations of other metabolites will not. If desired, this elasticity analysis can be repeated with the primary target blocks subdivided into simpler blocks so that the primary sites of action can be defined with more and more precision until, with sufficient subdivision, they are mapped onto individual kinetic steps. Top-down elasticity analysis has been used to identify the targets of effectors of oxygen consumption in mitochondria, hepatocytes and thymocytes. Effectors include poisons such as cadmium and hormones such as triiodothyronine. However, the method is more general than this; in principle it can be applied to any metabolic or other steady-state system. PMID- 9746310 TI - A model of O2.-generation in the complex III of the electron transport chain. AB - Oxidation of semiquinone by O2 in the Q cycle is known to be one of the sources of superoxide anion (O2.-) in aerobic cells. In this paper, such a phenomenon was analyzed using the chemical kinetics model of electron transfer from succinate to cytochrome c, including coenzyme Q, the complex III non-heme iron protein FeSIII and cytochromes bl, bh and cl. Electron transfers from QH2 to FeSIII and cytochrome bl were assumed to occur according to direct transfer mechanism (dynamic channelling) involving the formation of FeS(red)III-Q.- and Q.- cytochrome bl complexes. For oxidation/reduction reactions involving cytochromes bh and bl, the dependence of the equilibrium and elementary rate constants on the membrane potential (deltapsi) was taken into consideration. The rate of O2.- generation was found to increase dramatically with increase in deltapsi above the values found in State 3. On the other hand, the rate of cytochrome c reduction decreased sharply at the same values of the membrane potential. This explains experimental data that the O2.- generation at State 4 appears to be very much faster than at State 3. A mild uncoupling in State 4 can markedly decrease the superoxide generation due to a decrease in deltapsi below the above mentioned critical level. DeltapH appears to be equally effective as deltapsi in stimulation of superoxide production which depends, in fact, upon the deltamuH+ level. PMID- 9746312 TI - Oxidative phosphorylation in intact hepatocytes: quantitative characterization of the mechanisms of change in efficiency and cellular consequences. AB - Two mechanisms may affect the yield of the oxidative phosphorylation pathway in isolated mitochondria: (i) a decrease in the intrinsic coupling of the proton pumps (H+/2e- or H+/ATP), and (ii) an increase in the inner membrane conductance (proton or cation leak). Hence three kinds of modifications can occur and each of them have been characterized in isolated rat liver mitochondria (see preceding chapter by Rigoulet et al.). In intact isolated hepatocytes, these modifications are linked to specific patterns of bioenergetic parameters, i.e. respiratory flux, mitochondrial redox potential, DY, and phosphate potential. (1) The increase in H+/ATP stoichiometry of the mitochondrial ATP synthase, as induced by almitrine [20], leads to a decrease in mitochondrial and cytosolic ATP/ADP ratios without any change in the protonmotive force nor in the respiratory rate or redox potential. (2) In comparison to carbohydrate, octanoate metabolism by beta oxidation increases the proportion of electrons supplied at the second coupling site of the respiratory chain. This mimics a redox slipping. Octanoate addition results in an increased respiratory rate and mitochondrial NADH/NAD ratio while protonmotive force and phosphate potential are almost unaffected. The respiratory rate increase is associated with a decrease in the overall apparent thermodynamic driving force (2deltaE'o - ndeltap) which confirms the 'redox-slipping-like' effect. (3) An increase in proton conductance as induced by the protonophoric uncoupler 2,4-dinitrophenol (DNP) leads to a decrease, as expected, in the mitochondrial NADH/NAD and ATP/ ADP ratios and in deltapsi while respiratory rate is increased. Thus, each kind of modification (proton leak, respiratory chain redox slipping or increase in H+/ATP stoichiometry of ATPase) is related to a specific set of bioenergetic parameters in intact cells. Moreover, these patterns are in good agreement with the data found in isolated mitochondria. From this work, we conclude that quantitative analysis of four bioenergetic parameters (respiration rate, mitochondrial NADH/ NAD ratio, protonmotive force and mitochondrial phosphate potential) gives adequate tools to investigate the mechanism by which some alterations may affect the yield of the oxidative phosphorylation pathway in intact cells. PMID- 9746311 TI - Quantitative analysis of some mechanisms affecting the yield of oxidative phosphorylation: dependence upon both fluxes and forces. AB - The purpose of this work was to show how the quantitative definition of the different parameters involved in mitochondrial oxidative phosphorylation makes it possible to characterize the mechanisms by which the yield of ATP synthesis is affected. Three different factors have to be considered: (i) the size of the different forces involved (free energy of redox reactions and ATP synthesis, proton electrochemical difference); (ii) the physical properties of the inner mitochondrial membrane in terms of leaks (H+ and cations); and finally (iii) the properties of the different proton pumps involved in this system (kinetic properties, regulation, modification of intrinsic stoichiometry). The data presented different situations where one or more of these parameters are affected, leading to a different yield of oxidative phosphorylation. (1) By manipulating the actual flux through each of the respiratory chain units at constant protonmotive force in yeast mitochondria, we show that the ATP/O ratio decreases when the flux increases. Moreover, the highest efficiency was obtained when the respiratory rate was low and almost entirely controlled by the electron supply. (2) By using almitrine in different kinds of mitochondria, we show that this drug leads to a decrease in ATP synthesis efficiency by increasing the H+/ATP stoichiometry ofATP synthase (Rigoulet M et al. Biochim Biophys Acta 1018: 91-97, 1990). Since this enzyme is reversible, it was possible to test the effect of this drug on the reverse reaction of the enzyme i.e. extrusion of protons catalyzed by ATP hydrolysis. Hence, we are able to prove that, in this case, the decrease in efficiency of oxidative phosphorylation is due to a change in the mechanistic stoichiometry of this proton pump. To our knowledge, this is the first example of a modification in oxidative phosphorylation yield by a change in mechanistic stoichiometry of one of the proton pumps involved. (3) In a model of polyunsaturated fatty acid deficiency in rat, it was found that non-ohmic proton leak was increased, while ohmic leak was unchanged. Moreover, an increase in redox slipping was also involved, leading to a complex picture. However, the respective role of these two mechanisms may be deduced from their intrinsic properties. For each steady state condition, the quantitative effect of these two mechanisms in the decrease of oxidative phosphorylation efficiency depends on the values of different fluxes or forces involved. (4) Finally the comparison of the thermokinetic data in view of the three dimensional-structure of some pumps (X ray diffraction) also gives some information concerning the putative mechanism of coupling (i.e. redox loop or proton pump) and their kinetic control versus regulation of mitochondrial oxidative phosphorylation. PMID- 9746313 TI - Yeast mitochondrial metabolism: from in vitro to in situ quantitative study. AB - In this work, we first compared yeast mitochondrial oxidative metabolism at different levels of organization: whole cells (C), spheroplasts (S), permeabilized spheroplasts (PS) or isolated mitochondria (M). At present, S are more suitable for use than C for biochemical techniques such as fast extraction of metabolites and permeabilization. We show here that respiratory rates of S with various substrates are similar to C, which demonstrate that they are adapted to yeast bioenergetic studies. It appeared from ethanol metabolism +/- NAD+ or NADH respiratory rates on PS that ethanol metabolism was largely cytosolic; moreover, the activity of NADH dehydrogenase was lesser in the case of PS than in S. By comparing PS and M, the biggest difference concerned the respiratory rates of pyruvate and pyruvate-malate, which were much lower for M. Thus mitochondria preparation caused an unidentified loss involved directly in pyruvate metabolism. When the respiratory rate was lowered as a consequence of a high kinetic control of oxidative activity upstream from the respiratory chain, a similar correlation between the increase in ATP/O and decrease in respiratory rate was observed. So, the intrinsic uncoupling of proton pumps is not a particularity of M. Secondly, we demonstrate the existence of a mechanism of retarded diffusion in yeast similar to that already observed in permeabilized mammalian cells for ADP. Such a mechanism also occurs in yeast for several respiratory substrates: the K0.5 for each substrate toward the respiration rate in PS always exceeds that for M. It is proposed that such a discrepancy is due to a restriction of metabolite movement across the outer mitochondrial membrane in permeabilized cells, i.e. regulation of the substrate permeability through porin channels. In the porin-deficient yeast mutant, the K0.5 for NADH is not significantly different in either M or PS and is comparable to that of the parent strain PS. This result confirms that this retarded diffusion is essentially due to the opening-closing of the porin channel. PMID- 9746314 TI - Permeabilized cell and skinned fiber techniques in studies of mitochondrial function in vivo. AB - In this chapter we describe in details the permeabilized cell and skinned fiber techniques and their applications for studies of mitochondrial function in vivo. The experience of more than 10 years of research in four countries is summarized. The use of saponin in very low concentration (50-100 microg/ml) for permeabilisation of the sarcolemma leaves all intracellular structures, including mitochondria, completely intact. The intactness of mitochondrial function in these skinned muscle fibers is demonstrated in this work by multiple methods, such as NADH and flavoprotein fluorescence studies, fluorescence imaging, confocal immunofluorescence microscopy and respiratory analysis. Permeabilized cell and skinned fiber techniques have several very significant advantages for studies of mitochondrial function, in comparison with the traditional methods of use of isolated mitochondria: (1) very small tissue samples are required; (2) all cellular population of mitochondria can be investigated; (3) most important, however, is that mitochondria are studied in their natural surrounding. The results of research by using this method show the existence of several new phenomenon--tissue dependence of the mechanism of regulation of mitochondrial respiration, and activation of respiration by selective proteolysis. These phenomena are explained by interaction of mitochondria with other cellular structures in vivo. The details of experimental studies with use of these techniques and problems of kinetic analysis of the results are discussed. Examples of large-scale clinical application of these methods are given. PMID- 9746315 TI - Cytoskeleton and mitochondrial morphology and function. AB - It has been well established that the cytoskeleton is an essential modulator of cell morphology and motility, intracytoplasmic transport and mitosis, however cytoskeletal linkage to the organelles has not been unequivocally demonstrated. Indeed, cytoskeleton appears to be essential in determining and modulating gene phenotype as a function of cellular environment. According to recent studies, the organization of the cytoskeleton network together with associated protein(s) could be essential in regulating mitochondrial function and particularly the permeability of the mitochondrial outer membrane to ADP. The aim of this chapter is to summarize the main properties of the cytoskeletal environment of mitochondria and the possible role(s) of this network in mitochondrial function in myocytes. PMID- 9746316 TI - Energetics of swelling in isolated hepatocytes: a comprehensive study. AB - Cell swelling is now admitted as being a new principle of metabolic control but little is known about the energetics of cell swelling. We have studied the influence of hypo- or hyperosmolarity on both isolated hepatocytes and isolated rat liver mitochondria. Cytosolic hypoosmolarity on isolated hepatocytes induces an increase in matricial volume and does not affect the myxothiazol sensitive respiratory rate while the absolute value of the overall thermodynamic driving force over the electron transport chain increases. This points to an increase in kinetic control upstream the respiratory chain when cytosolic osmolarity is decreased. On isolated rat liver mitochondria incubated in hypoosmotic potassium chloride media, energetic parameters vary as in cells and oxidative phosphorylation efficiency is not affected. Cytosolic hyperosmolarity induced by sodium co-transported amino acids, per se, does not affect either matrix volume or energetic parameters. This is not the case in isolated rat liver mitochondria incubated in sucrose hyperosmotic medium. Indeed, in this medium, adenine nucleotide carrier is inhibited as the external osmolarity increases, which lowers the state 3 respiration close to state 4 level and consequently leads to a decrease in oxidative phosphorylation efficiency. When isolated rat liver mitochondria are incubated in KCl hyperosmotic medium, state 3 respiratory rate, matrix volume and membrane electrical potential vary as a function of time. Indeed, matrix volume is recovered in hyperosmotic KCl medium and this recovery is dependent on Pi-Kentry. State 3 respiratory rate increases and membrane electrical potential difference decreases during the first minutes of mitochondrial incubation until the attainment of the same value as in isoosmotic medium. This shows that matrix volume, flux and force are regulated as a function of time in KCl hyperosmotic medium. Under steady state, neither matrix volume nor energetic parameters are affected. Moreover, NaCl hyperosmotic medium allows matrix volume recovery but induces a decrease in state 3 respiratory flux. This indicates that potassium is necessary for both matrix volume and flux recovery in isolated mitochondria. We conclude that hypoosmotic medium induces an increase in kinetic control both upstream and on the respiratory chain and changes the oxidative phosphorylation response to forces. At steady state, hyperosmolarity, per se, has no effect on oxidative phosphorylation in either isolated hepatocytes or isolated mitochondria incubated in KCl medium. Therefore, potassium plays a key role in matrix volume, flux and force regulation. PMID- 9746317 TI - Functional aspects of the X-ray structure of mitochondrial creatine kinase: a molecular physiology approach. AB - Mitochondrial creatine kinase (Mi-CK) is a central enzyme in energy metabolism of tissues with high and fluctuating energy requirements. In this review, recent progress in the functional and structural characterization of Mi-CK is summarized with special emphasis on the solved X-ray structure of chicken Mib-CK octamer (Fritz-Wolf et al., Nature 381, 341-345, 1996). The new results are discussed in a historical context and related to the characteristics of CK isoforms as known from a large number of biophysical and biochemical studies. Finally, two hypothetical functional aspects of the Mi-CK structure are proposed: (i) putative membrane binding motifs at the top and bottom faces of the octamer and (ii) a possible functional role of the central 20 A channel. PMID- 9746318 TI - Oligomeric state and membrane binding behaviour of creatine kinase isoenzymes: implications for cellular function and mitochondrial structure. AB - The membrane binding properties of cytosolic and mitochondrial creatine kinase isoenzymes are reviewed in this article. Differences between both dimeric and octameric mitochondrial creatine kinase (Mi-CK) attached to membranes and the unbound form are elaborated with respect to possible biological function. The formation of crystalline mitochondrial inclusions under pathological conditions and its possible origin in the membrane attachment capabilities of Mi-CK are discussed. Finally, the implications of these results on mitochondrial energy transduction and structure are presented. PMID- 9746319 TI - Molecular characterization of the creatine kinases and some historical perspectives. AB - Over the last 15 years, molecular characterization of the creatine kinase (CK) gene family has paralleled the molecular revolution of understanding gene structure, function, and regulation. In this review, we present a summary of advances in molecular analysis of the CK gene family with a few vignettes of historical interest. We describe how the muscle CK gene provided an essential model system to examine myogenic regulatory mechanisms, leading to the discovery of the binding site for the MyoD family of basic helix-loop-helix transcription factors essential in skeletal myogenesis and the characterization of the MEF2 family of factors with an A/T rich consensus binding site essential in skeletal myogenesis and cardiogenesis. Cloning and characterization of the four mRNAs and nuclear genes encoding the cytosolic CKs, muscle and brain CKs, and the mitochondrial (Mt) CKs, sarcomeric MtCK and ubiquitous MtCK, has allowed intriguing study of tissue-specific and cell-specific expression of the different CKs and analysis of structural, functional, regulatory, and evolutionary relationships among both the four CK proteins and genes. Current and future studies focus on understanding both cellular energetics facilitated by the CK enzymes, especially energy channelling from the site of production, the mitochondrial matrix and inner membrane, to various cytosolic foci of utilization, and regulation of MtCK gene expression at the cell and tissue specific level as models of regulation of energy producing genes. PMID- 9746320 TI - Adenylate kinase: kinetic behavior in intact cells indicates it is integral to multiple cellular processes. AB - Monitoring the kinetic behavior of adenylate kinase (AK) and creatine kinase (CK) in intact cells by 18O-phosphoryl oxygen exchange analysis has provided new perspectives from which to more fully define the involvement of these phosphotransferases in cellular bioenergetics. A primary function attributable to both AK and CK is their apparent capability to couple ATP utilization with its generation by glycolytic and/or oxidative processes depending on cell metabolic status. This is evidenced by the observation that the sum of the net AK- plus CK catalyzed phosphoryl transfer is equivalent to about 95% of the total ATP metabolic flux in non-contracting rat diaphragm; under basal conditions almost every newly generated ATP molecule appears to be processed by one or the other of these phosphotransferases prior to its utilization. Although CK accounts for the transfer of a majority of the ATP molecules generated/consumed in the basal state there is a progressive, apparently compensatory, shift in phosphotransfer catalysis from the CK to the AK system with increasing muscle contraction or graded chemical inhibition of CK activity. AK and CK appear therefore to provide similar and interrelated functions. Evidence that high energy phosphoryl transfer in some cell types or metabolic states can also be provided by specific nucleoside mono- and diphosphate kinases and by the phosphotransfer capability inherent to the glycolytic system has been obtained. Measurements by 18O-exchange analyses of net AK- and CK-catalyzed phosphoryl transfer in conjunction with 31P NMR analyses of total unidirectional phosphoryl flux show that each new energy bearing molecule CK or AK generates subsequently undergoes about 50 or more unidirectional CK-or AK-catalyzed phosphotransfers en route to an ATP consumption site in intact muscle. This evidence of multiple enzyme catalyzed exchanges coincides with the mechanism of vectorial ligand conduction suggested for accomplishing intracellular high energy phosphoryl transfer by the AK and CK systems. AK-catalyzed phosphotransfer also appears to be integral to the transduction of metabolic signals influencing the operation of ion channels regulated by adenine nucleotides such as ATP-inhibitable K+ channels in insulin secreting cells; transition from the ATP to ADP liganded states closely coincides with the rate AK-catalyzes phosphotransfer transforming ATP (+AMP) to (2)ADP. PMID- 9746321 TI - Cytoarchitectural and metabolic adaptations in muscles with mitochondrial and cytosolic creatine kinase deficiencies. AB - We have blocked creatine kinase (CK) mediated phosphocreatine (PCr) <==> ATP transphosphorylation in mitochondria and cytosol of skeletal muscle by knocking out the genes for the mitochondrial (ScCKmit) and the cytosolic (M-CK) CK isoforms in mice. Animals which carry single or double mutations, if kept and tested under standard laboratory conditions, have surprisingly mild changes in muscle physiology. Strenuous ex vivo conditions were necessary to reveal that MM CK absence in single and double mutants leads to a partial loss of tetanic force output. Single ScCKmit deficiency has no noticeable effects but in combination the mutations cause slowing of the relaxation rate. Importantly, our studies revealed that there is metabolic and cytoarchitectural adaptation to CK defects in energy metabolism. The effects involve mutation type-dependent alterations in the levels of AMP, IMP, glycogen and phosphomonoesters, changes in activity of metabolic enzymes like AMP-deaminase, alterations in mitochondrial volume and contractile protein (MHC isoform) profiles, and a hyperproliferation of the terminal cysternae of the SR (in tubular aggregates). This suggests that there is a compensatory resiliency of loss-of-function and redirection of flux distributions in the metabolic network for cellular energy in our mutants. PMID- 9746322 TI - In situ measurements of creatine kinase flux by NMR. The lessons from bioengineered mice. AB - P-31 nuclear magnetic resonance (NMR) is uniquely suited to measure the kinetics of the phosphoryl-exchange reaction catalyzed by creatine kinase in intact mammalian tissue, especially striated muscle. Recently developed transgenic mouse models of the creatine kinase iso-enzyme system open novel opportunities to assess the functional importance of the individual iso-enzymes and their relative contribution to the total in situ flux through the CK reaction. This chapter reviews the most recent findings from NMR flux measurements on such genetic models of CK function. Findings in intact mouse skeletal and cardiac muscle in vivo are compared to data from purified mitochondrial and cytosolic creatine kinase in vitro. The relevance of findings in transgenic animals for the function of CK in wild-type tissue is described and the perspectives of transgenic techniques in future quantitative studies on the creatine kinase iso-enzyme system are indicated. PMID- 9746323 TI - Mathematical model of compartmentalized energy transfer: its use for analysis and interpretation of 31P-NMR studies of isolated heart of creatine kinase deficient mice. AB - A mathematical model of the compartmentalized energy transfer in cardiac cells is described and used for interpretation of novel experimental data obtained by using phosphorus NMR for determination of the energy fluxes in the isolated hearts of transgenic mice with knocked out creatine kinase isoenzymes. These experiments were designed to study the meaning and importance of compartmentation of creatine kinase isoenzymes in the cells in vivo. The model was constructed to describe quantitatively the processes of energy production, transfer, utilization, and feedback between these processes. It describes the production of ATP in mitochondrial matrix space by ATP synthase, use of this ATP for phosphocreatine production in the mitochondrial creatine kinase reaction coupled to the adenine nucleotide translocation, diffusional exchange of metabolites in the cytoplasmic space, and use of phosphocreatine for resynthesis of ATP in the myoplasmic creatine kinase reaction. It accounts also for the recently discovered phenomenon of restricted diffusion of adenine nucleotides through mitochondrial outer membrane porin pores (VDAC). Practically all parameters of the model were determined experimentally. The analysis of energy fluxes between different cellular compartments shows that in all cellular compartments of working heart cells the creatine kinase reaction is far from equilibrium in the systolic phase of the contraction cycle and approaches equilibrium only in cytoplasm and only in the end-diastolic phase of the contraction cycle. Experimental determination of the relationship between energy fluxes by a 31P-NMR saturation transfer method and workload in isolated and perfused heart of transgenic mice deficient in MM isoenzyme of the creatine kinase, MM-/-showed that in the hearts from wild mice, containing all creatine kinase isoenzymes, the energy fluxes determined increased 3-4 times with elevation of the workload. By contrast, in the hearts in which only the mitochondrial creatine kinase was active, the energy fluxes became practically independent of the workload in spite of the preservation of 26% of normal creatine kinase activity. These results cannot be explained on the basis of the conventional near-equilibrium theory of creatine kinase in the cells, which excludes any difference between creatine kinase isoenzymes. However, these apparently paradoxical experimental results are quantitatively described by a mathematical model of the compartmentalized energy transfer based on the steady state kinetics of coupled creatine kinase reactions, compartmentation of creatine kinase isoenzymes in the cells, and the kinetics of ATP production and utilization reactions. The use of this model shows that: (1) in the wild type heart cells a major part of energy is transported out of mitochondria via phosphocreatine, which is used for complete regeneration of ATP locally in the myofibrils--this is the quantitative estimate for PCr pathway; (2) however, in the absence of MM-creatine kinase in the myofibrils in transgenic mice the contraction results in a very rapid rise of ADP in cytoplasmic space, that reverses the mitochondrial creatine kinase reaction in the direction of ATP production. In this way, because of increasing concentrations of cytoplasmic ADP, mitochondrial creatine kinase is switched off functionally due to the absence of its counterpart in PCr pathway, MM-creatine kinase. This may explain why the creatine kinase flux becomes practically independent from the workload in the hearts of transgenic mouse without MM-CK. Thus, the analysis of the results of studies of hearts of creatine kinase-deficient transgenic mice, based on the use of a mathematical model of compartmentalized energy transfer, show that in the PCr pathway of intracellular energy transport two isoenzymes of creatine kinase always function in a coordinated manner out of equilibrium, in the steady state, and disturbances in functioning of one of them inevitably result PMID- 9746324 TI - Functional coupling of creatine kinases in muscles: species and tissue specificity. AB - Creatine kinase (CK) isoenzymes are present in all vertebrates. An important property of the creatine kinase system is that its total activity, its isoform distribution, and the concentration of guanidino substrates are highly variable among species and tissues. In the highly organized structure of adult muscles, it has been shown that specific CK isoenzymes are bound to intracellular compartments, and are functionally coupled to enzymes and transport systems involved in energy production and utilization. It is however, not established whether functional coupling and intracellular compartmentation are present in all vertebrates. Furthermore, these characteristics seem to be different among different muscle types within a given species. This study will review some of these aspects. It has been observed that: (1) In heart ventricle, CK compartmentation and coupling characterize adult mammalian cells. It is almost absent in frogs, and is weakly present in birds. (2) Efficient coupling of MM-CK to myosin ATPase is seen in adult mammalian striated muscles but not in frog and bird heart where B-CK is expressed instead of M-CK. Thus, the functional efficacy of bound MM-CK to regulate adenine nucleotide turnover within the myofibrillar compartment seems to be specific for muscles expressing M-CK as an integral part of the sarcomere. (3) Mi-CK expression and/or functional coupling are highly tissue and species specific; moreover, they are subject to short term and long term adaptations, and are present late in development. The mitochondrial form of CK (mi-CK) can function in two modes depending on the tissue: (i) in an <> and (ii) in an <>. The mode of action of mi-CK seems to be related to its precise localization within the mitochondrial intermembrane space, whereas its amount might control the quantitative aspects of the coupling. Mi-CK is highly plastic, making it a strong candidate for fine regulation of excitation-contraction coupling in muscles and for energy transfer in cells with large and fluctuating energy demands in general. (4) Although CK isoforms show a binding specificity, the presence of a given isoform within a tissue or a species only, does not predict its functional role. For example, M-CK is expressed before it is functionally compartmentalized within myofibrils during development. Similarly, the presence of ubiquitous or sarcomeric mi-CK isoforms, is not an index of functional coupling of mi-CK to oxidative phosphorylation. (5) Amongst species or muscles, it appears that a large buffering action of the CK system is associated with rapid contraction and high glycolytic activity. On the other hand, an oxidative metabolism is associated with isoform diversity, increased compartmentation, a subsequent low buffering action and efficient phosphotransfer between mitochondria and energy utilization sites. It can be concluded that, in addition to a high variation of total activity and isoform expression, the role of the CK system also critically depends on its intracellular organization and interaction with energy producing and utilizing pathways. This compartmentation will determine the high cellular efficiency and fine specialization of highly organized and differentiated muscle cells. PMID- 9746326 TI - Quantitative studies of enzyme-substrate compartmentation, functional coupling and metabolic channelling in muscle cells. AB - Some historical aspects of development of the concepts of functional coupling, metabolic channelling, compartmentation and energy transfer networks are reviewed. Different quantitative approaches, including kinetic and mathematical modeling of energy metabolism, intracellular energy transfer and metabolic regulation of energy production and fluxes in the cells in vivo are analyzed. As an example of the system with metabolic channelling, thermodynamic aspects of the functioning the mitochondrial creatine kinase functionally coupled to the oxidative phosphorylation are considered. The internal thermodynamics of the mitochondrial creatine kinase reaction is similar to that for other isoenzymes of creatine kinase, and the oxidative phosphorylation process specifically influences steps of association and dissociation of MgATP with the enzyme due to channelling of ATP from adenine nucleotide translocase. A new paradigm of muscle bioenergetics-the paradigm of energy transfer and feedback signaling networks based on analysis of compartmentation phenomena and structural and functional interactions in the cell is described. Analysis of the results of mathematical modeling of the compartmentalized energy transfer leads to conclusion that both calcium and ADP, which concentration changes synchronously in contraction cycle, may simultaneously activate oxidative phosphorylation in the muscle cells in vivo. The importance of the phosphocreatine circuit among other pathways of intracellular energy transfer network is discussed on the basis of the recent data published in the literature, with some experimental demonstration. The results of studies of perfused rat hearts with completely inhibited creatine kinase show significantly decreased work capacity and respectively, energy fluxes, in these hearts in spite of significant activation of adenylate kinase system (Dzeja et al. this volume). These results, combined with those of mathematical analysis of the energy metabolism of hearts of transgenic mice with switched off creatine kinase isoenzymes confirm the importance of phosphocreatine pathway for energy transfer for cell function and energetics in mature heart and many other types of cells, as one of major parts of intracellular energy transfer network and metabolic regulation. PMID- 9746325 TI - Theoretical modelling of some spatial and temporal aspects of the mitochondrion/creatine kinase/myofibril system in muscle. AB - After discussing approaches to the modelling of mitochondrial regulation in muscle, we describe a model that takes account, in a simplified way, of some aspects of the metabolic and physical structure of the energy production/usage system. In this model, high-energy phosphates (ATP and phosphocreatine) and low energy metabolites (ADP and creatine) diffuse between the mitochondrion and the myofibrillar ATPase, and can be exchanged at any point by creatine kinase. Creatine kinase is not assumed to be at equilibrium, so explicit account can be taken of substantial changes in its activity of the sort that can now be achieved by transgenic technology in vivo. The ATPase rate is the input function. Oxidative ATP synthesis is controlled by juxtamitochondrial ADP concentration. To allow for possible functional 'coupling' between the components of creatine kinase associated with the mitochondrial adenine nucleotide translocase and the myofibrillar ATPase, we define parameters phi and psi that set the fraction of the total flux carried by ATP rather than phosphocreatine out of the mitochondrial unit and into the ATPase unit, respectively. This simplification is justified by a detailed analysis of the interplay between the mitochondrial outer membrane porin proteins, mitochondrial creatine kinase and the adenine nucleotide translocase. As both processes of possible 'coupling' are incorporated into the model as quantitative parameters, their effect on the energetics of the whole cell model can be explicitly assessed. The main findings are as follows: (1) At high creatine kinase activity, the hyperbolic relationship of oxidative ATP synthesis rate to spatially averaged ADP concentration at steady state implies also a near-linear relationship to creatine concentration, and a sigmoid relation to free energy of ATP hydrolysis. At high creatine kinase activity, the degree of functional coupling at either the mitochondrial or ATPase end has little effect on these relationships. However, lowering the creatine kinase activity raises the mean steady state ADP and creatine concentrations, and this is exaggerated when phi or psi is near unity (i.e. little coupling). (2) At high creatine kinase activity, the fraction of flow at steady state carried in the middle of the model by ATP is small, unaffected by the degree of functional coupling, but increases with ADP concentration and rate of ATP turnover. Lowering the creatine kinase activity raises this fraction, and this is exaggerated when psi or psi is near unity. (3) Both creatine and ADP concentrations show small gradients decreasing towards the mitochondrion (in the direction of their net flux), while ATP and phosphocreatine concentration show small gradients decreasing towards the myosin ATPase. Unless phi = psi = 0 (i.e. complete coupling), there is a gradient of net creatine kinase flux that results from the need to transform some of the 'adenine nucleotide flux' at the ends of the model into 'creatine flux' in the middle; the overall net flux is small, but only zero if phi = psi. A reduction in cytosolic creatine kinase activity decreases ADP concentration at the mitochondrial end and increases it at the ATPase end. (4) During work-jump transitions, spatial average responses exhibit exponential kinetics similar to those of models of mitochondrial control that assume equilibrium conditions for creatine kinase. (5) In response to a step increase in ATPase activity, concentration changes start at the ATPase end and propagate towards the mitochondrion, damped in time and space. This simplified model embodies many important features of muscle in vivo, and accommodates a range of current theories as special cases. We end by discussing its relationship to other approaches to mitochondrial regulation in muscle, and some possible extensions of the model. PMID- 9746327 TI - Subtleties in control by metabolic channelling and enzyme organization. AB - Because of its importance to cell function, the free-energy metabolism of the living cell is subtly and homeostatically controlled. Metabolic control analysis enables a quantitative determination of what controls the relevant fluxes. However, the original metabolic control analysis was developed for idealized metabolic systems, which were assumed to lack enzyme-enzyme association and direct metabolite transfer between enzymes (channelling). We here review the recently developed molecular control analysis, which makes it possible to study non-ideal (channelled, organized) systems quantitatively in terms of what controls the fluxes, concentrations, and transit times. We show that in real, non ideal pathways, the central control laws, such as the summation theorem for flux control, are richer than in ideal systems: the sum of the control of the enzymes participating in a non-ideal pathway may well exceed one (the number expected in the ideal pathways), but may also drop to values below one. Precise expressions indicate how total control is determined by non-ideal phenomena such as ternary complex formation (two enzymes, one metabolite), and enzyme sequestration. The bacterial phosphotransferase system (PTS), which catalyses the uptake and concomitant phosphorylation of glucose (and also regulates catabolite repression) is analyzed as an experimental example of a non-ideal pathway. Here, the phosphoryl group is channelled between enzymes, which could increase the sum of the enzyme control coefficients to two, whereas the formation of ternary complexes could decrease the sum of the enzyme control coefficients to below one. Experimental studies have recently confirmed this identification, as well as theoretically predicted values for the total control. Macromolecular crowding was shown to be a major candidate for the factor that modulates the non-ideal behaviour of the PTS pathway and the sum of the enzyme control coefficients. PMID- 9746329 TI - Is it possible to predict any properties of oxidative phosphorylation in a theoretical way? AB - Two theoretical approaches applied to oxidative phosphorylation, namely Metabolic Control Analysis (MCA) [ 1-7] and Non-Equilibrium Thermodynamics (NET) [8-11], turned out to be very useful tools for quantitative description and understanding of control and regulation of this process. However, they were not able to predict any new properties of the considered system. On the other hand, the previously developed dynamic model of oxidative phosphorylation [12-17], representing a kinetic approach, allowed to formulate several interesting predictions which can be tested experimentally. The most important of these predictions are: (1) Different steps of ATP-production must be directly activated to a similar extent as ATP-consumption during stimulation of ATP turnover by calcium-acting hormones as well as by neural signals during muscle contraction; (2) A universal activator/regulatory mechanism responsible for such a precise balance of activation should be identified; (3) The flux-force relationship for cytochrome oxidase can be inverse during the transition towards hypoxia and anoxia, when oxygen concentration falls below 30 microM; (4) The flux-force relationship can depend on the way in which the thermodynamic force is changed; (5) The pattern of metabolic control is completely different in normoxic and hypoxic conditions; in the latter case cytochrome oxidase has the flux control coefficient close to unity. Thus, the kinetic model of oxidative phosphorylation seems to be a useful scientific tool, offering some novel theoretical predictions, which then can be tested in the experimental way. PMID- 9746328 TI - The dynamic regulation of myocardial oxidative phosphorylation: analysis of the response time of oxygen consumption. AB - Although usually steady-state fluxes and metabolite levels are assessed for the study of metabolic regulation, much can be learned from studying the transient response during quick changes of an input to the system. To this end we study the transient response of O2 consumption in the heart during steps in heart rate. The time course is characterized by the mean response time of O2 consumption which is the first statistical moment of the impulse response function of the system (for mono-exponential responses equal to the time constant). The time course of O2 uptake during quick changes is measured with O2 electrodes in the arterial perfusate and venous effluent of the heart, but the venous signal is delayed with respect to O2 consumption in the mitochondria due to O2 diffusion and vascular transport. We correct for this transport delay by using the mass balance of O2, with all terms (e.g. O2 consumption and vascular O2 transport) taken as function of time. Integration of this mass balance over the duration of the response yields a relation between the mean transit time for O2 and changes in cardiac O2 content. Experimental data on the response times of venous [O2] during step changes in arterial [O2] or in perfusion flow are used to calculate the transport time between mitochondria and the venous O2 electrode. By subtracting the transport time from the response time measured in the venous outflow the mean response time of mitochondrial O2 consumption (tmito) to the step in heart rate is obtained. In isolated rabbit heart we found that tmito to heart rate steps is 4-12 s at 37 degrees C. This means that oxidative phosphorylation responds to changing ATP hydrolysis with some delay, so that the phosphocreatine levels in the heart must be decreased, at least in the early stages after an increase in cardiac ATP hydrolysis. Changes in ADP and inorganic phosphate (Pi) thus play a role in regulating the dynamic adaptation of oxidative phosphorylation, although most steady state NMR measurements in the heart had suggested that ADP and Pi do not change. Indeed, we found with 31P-NMR spectroscopy that phosphocreatine (PCr) and Pi change in the first seconds after a quick change in ATP hydrolysis, but remarkably they do this significantly faster (time constant approximately 2.5 s) than mitochondrial O2 consumption (time constant 12 s). Although it is quite likely that other factors besides ADP and Pi regulate cardiac oxidative phosphorylation, a fascinating alternative explanation is that the first changes in PCr measured with NMR spectroscopy took exclusively place in or near the myofibrils, and that a metabolic wave must then travel with some delay to the mitochondria to stimulate oxidative phosphorylation. The tmito slows with falling temperature, intracellular acidosis, and sometimes also during reperfusion following ischemia and with decreased mitochondrial aerobic capacity. In conclusion, the study of the dynamic adaptation of cardiac oxidative phosphorylation to demand using the mean response time of cardiac mitochondrial O2 consumption is a very valuable tool to investigate the regulation of cardiac mitochondrial energy metabolism in health and disease. PMID- 9746331 TI - Modulation of cell calcium signals by mitochondria. AB - It is now clearer and clearer that mitochondria play a role, and perhaps an active role, in cell calcium signalling. The fact that mitochondria can exhibit a Ca2+-induced Ca2+ release (mCICR, Ichas et al. [37]) reinforces this concept and makes the mitochondria an essential element in the relay of Ca2+ wave propagation. It must be emphasized that the modulation of cell Ca2+ signals by mitochondria depends upon their energetic status, thus making mitochondria an essential link between energy metabolism and calcium signalling inside the cell. PMID- 9746330 TI - Role of mitochondrial calcium transport in the control of substrate oxidation. AB - This paper reviews the model of the control of mitochondrial substrate oxidation by Ca2+ ions. The mechanism is the activation by Ca2+ of four mitochondrial dehydrogenases, viz. glycerol 3-phosphate dehydrogenase, the pyruvate dehydrogenase multienzyme complex (PDH), NAD-linked isocitrate dehydrogenase (NAD IDH) and 2-oxoglutarate dehydrogenase (OGDH). This results in the increase, or near-maintenance, of mitochondrial NADH/NAD ratios in the activated state, depending upon the tissue and the degree of 'downstream' activation by Ca2+, likely at the level of the F1Fo ATPase. Higher values of the redox span of the respiratory chain allow for greatly increased fluxes through oxidative phosphorylation with a minimal drop in protonmotive force and phosphorylation potential. As PDH, NAD-IDH and OGDH are all located within the inner mitochondrial membrane, it is changes in matrix free Ca2+ [Ca2+]m which act as a signal to these activities. In this article, we review recent work in which [Ca2+]m is measured in cells and tissues, using different techniques, with special emphasis on the question of the degree of damping of [Ca2+]m relative to changes in cytosol free Ca2+ in cells with rapid transients in cytosol Ca2+, e.g. cardiac myocytes. Further, we put forward the point of view that the failure of mitochondrial energy transduction to keep pace with cellular energy needs in some forms of heart failure may involve a failure of [Ca2+]m to be raised adequately to allow the activation of the dehydrogenases. We present new data to show that this is so in cardiac myocytes isolated from animals suffering from chronic, streptozocin-induced diabetes. This raises the possibility of therapy based upon partial inhibition of mitochondrial Ca2+ efflux pathways, thereby raising [Ca2+]m at a given, time-average value of cytosol free Ca+2. PMID- 9746333 TI - Role of cellular energetics in ischemia-reperfusion and ischemic preconditioning of myocardium. AB - A short period of ischemia followed by reperfusion produces a state of affairs in which the cells' potential for surviving longer ischemia is enhanced. This is called ischemic preconditioning. The effects of preconditioning are also related to the reperfusion damage which ensues upon tissue oxygenation. The role of the cellular energy state in reperfusion damage remains an enigma, although ischemic preconditioning is known to trigger mechanisms which contribute to the prevention of unnecessary ATP waste. In some species up to 80% of ATP hydrolysis in ischemia can be attributed to mitochondrial F1-F0-ATPase (ATP synthase), and a role for its inhibitor protein (IF1) in ATP preservation has been proposed. Although originally regarded as limited to large animals with a slow heart beat, inhibition by IF1 is probably a universal phenomenon. Coincidentally with ATPase inhibition, the decline in cellular ATP slows down, but even so the difference in ATP concentration between preconditioned and non-conditioned hearts is still small at the final stages of a long ischemia, when the beneficial effect of preconditioning is observable, although the energy state during reperfusion remains low in hearts which do not recover. PMID- 9746332 TI - Mitochondrial function as a determinant of recovery or death in cell response to injury. AB - Many pathological conditions can be the cause or the consequence of mitochondrial dysfunction. For instance anoxia, which is initiated by a critical reduction of oxygen availability for mitochondrial oxidations, is followed by a wide variety of mitochondrial alterations. A crucial role in the evolution of cell injury is to be attributed to the direction of operation of the F0F1 ATPase, which may turn mitochondria into the major consumers of cellular ATP in the futile attempt to restore the proton electrochemical gradient. On the other hand, functional mitochondria can paradoxically accelerate or exacerbate cell damage. This concept is particularly relevant for the ischemic myocardium. Indeed, inhibition of the respiratory chain or addition of uncouplers of oxidative phosphorylation can both limit the extent of enzyme release in the intact heart and prevent the onset of irreversible morphological changes in isolated myocytes. From studies on different tissues in a variety of pathological conditions a general consensus emerges on the role of intracellular Ca2+ overload as a pivotal link between cellular alterations and mitochondrial dysfunction. Oxidative phosphorylation is reduced by a massive mitochondrial uptake of Ca2+, resulting in a vicious cycle whereby the reduced ATP availability is followed by a failure of the mechanisms which extrude Ca2+ from the sarcoplasm. In addition, the rise in [Ca2+]i could promote opening of the cyclosporin-sensitive mitochondrial permeability transition pore, leading to a sudden deltapsi(m) dissipation. Here, we review the changes in intracellular and intramitochondrial ionic homeostasis occurring during ischemia and reperfusion. In particular, we evaluate the potential contribution of the permeability transition pore to cellular damage and discuss the mechanisms which can determine the cellular fate from a mitochondrial point of view. PMID- 9746334 TI - Early ischemia-induced alterations of the outer mitochondrial membrane and the intermembrane space: a potential cause for altered energy transfer in cardiac muscle? AB - Our aim was to carefully analyse the time-dependent changes that affect the mitochondrial function of myocardial cells during and after an ischemic episode. To this end, variables characterizing mitochondrial function have been evaluated on myocardial samples from isolated rat hearts subjected to different conditions of ischemia. The technique of permeabilized fibers was used in order to evaluate the mitochondrial function whilst retaining intracellular structure. The earliest alteration that could be detected was a decrease in the stimulatory effect of creatine on mitochondrial respiration. This alteration became more pronounced as the severity (or duration) of the ischemia increased. Afterwards, a significant decrease in the apparent Km of mitochondrial respiration for ADP also appeared, followed by a diminution of the maximal respiration rate which was partly restored by adding cytochrome c. Finally, for the most severe conditions of ischemia, the basal respiratory rate also increased. These observations are indicative of a sequence of alterations affecting first the intermembrane space, then the outer mitochondrial membrane, and finally the inner membrane. The discussion is focused on the very early alterations, that could not be detected using the conventional techniques of isolated mitochondria. We postulate that these alterations to the intermembrane space and outer mitochondrial membrane can induce disturbances both in the channelling of energy from the mitochondria, and on the signalling towards the mitochondria. The potential consequences on the regulation of the production of energy (ATP, PC) by the mitochondria are evoked. PMID- 9746335 TI - Metabolic control analysis and mitochondrial pathologies. AB - One of the main salient features recognized in mitochondrial diseases is the existence of a threshold in the degree of a mitochondrial deficit for the expression of the disease. When expressed as a function of the degree of heteroplasmy, the value of the threshold can be very high, around 90% (mutated DNA/total DNA). This means that 10% of normal DNA is enough to sustain a quasi normal mitochondrial oxidative phosphorylating flux. We have shown that most of the compensation is done at the metabolic level: for instance a 70% deficit of cytochrome oxidase decreases the oxidative flux by just 10%. Similar patterns are observed for the other complexes. Using Metabolic Control Analysis (MCA), we have shown that this kind of result is inescapable: the threshold value can be correlated to the control coefficient of the deficient step. The value of the threshold is reinforced by slight increases at the transcriptional and translational level as we show in a simple mathematical model. Finally we associate the threshold in the expression of a deficit, to the threshold in the energy demand of different tissues, in order to describe various patterns of onset of mitochondrial diseases (double threshold hypothesis). PMID- 9746338 TI - Clinical cardiac magnetic resonance spectroscopy--present state and future directions. AB - MR spectroscopy opens a window to the non-invasive evaluation of various aspects of cardiac metabolism. Experimentally, the method has extensively been used since 1970's. 31P-MR allows the registration of cardiac high-energy phosphate metabolism to non-invasively estimate the energetic state of the heart: ATP, phosphocreatine, inorganic phosphate, monophosphate esters and intracellular pH can all be quantitated. In conjunction with extracellular shift reagents such as [DyTTHA]3- or [TmDOTP]5-, 23Na- and 39K-MR allow the measurement of intra- and extra-cellular cation pools. 1H-MR spectroscopy allows the detection of a large number of metabolites such as, e.g. creatine, lactate, or carnitine. Human cardiac spectrocsopy has so far been confined to the 31P nucleus. Localization techniques (DRESS, ISIS, 3D-CSI etc.) are required to confine the acquired signal to the heart region. Relative quantification is straightforward (phosphocreatine/ATP ratio), absolute quantification (mM) is under development. Cardiac 31P-MR spectroscopy has research application in at least three clinical areas: (1) Coronary artery disease: A biochemical stress test for non-invasive ischemia detection (decrease of phosphocreatine with exercise) and viability assessment via quantification of ATP may become feasible. (2) Heart failure: The phosphocreatine/ATP ratio may provide an independent index for grading of heart failure, allow to monitor the longterm effects of different forms of drug therapy on cardiac energy metabolism in heart failure, and may also hold prognostic information on survival. (3) Valve disease: It is possible that the decrease of phosphocreatine/ATP can be used to guide the timing for the valve replacement. At the present time, no routine clinical applications can be defined for the use of human cardiac spectroscopy in patients with cardiac disease. However, the technique holds great potential for the future as a non-invasive approach to cardiac metabolism, and in coming years routine applications may become reality. PMID- 9746336 TI - Mechanisms of thyroid hormone control over sensitivity and maximal contractile responsiveness to beta-adrenergic agonists in atria. AB - This paper discusses the mechanisms of two basic effects of thyroid hormones on atrial responses to beta-adrenergic agonists, i.e. increased inotropic sensitivity and decreased maximal contractile responsiveness. The increased sensitivity of atria to beta-adrenergic agonists under thyroid hormones appears to be related to increases in beta-adrenoceptor density and Gs/Gi protein ratio, leading to activation of Gs-mediated pathway, but suppression of Gi-mediated pathway of adenylate cyclase regulation. Therefore, the i/c concentrations of cAMP and corresponding inotropic responses achieve their maximums at lower doses of beta-adrenergic agonist. Thyroid hormones also decrease the expression of phospholamban, but increase the expression of sarcoplasmic reticulum Ca2+-pump. As a result, the basal activity of sarcoplasmic reticulum Ca2+-pump increases, but its beta-adrenergic activation through phosphorylation of phospholamban decreases. It is suggested that these changes are causal for decreased maximal inotropic and lusitropic responses of atria to beta-adrenergic agonists. PMID- 9746337 TI - Creatine supplementation in health and disease. Effects of chronic creatine ingestion in vivo: down-regulation of the expression of creatine transporter isoforms in skeletal muscle. AB - Interest in creatine (Cr) as a nutritional supplement and ergogenic aid for athletes has surged over recent years. After cellular uptake, Cr is phosphorylated to phosphocreatine (PCr) by the creatine kinase (CK) reaction using ATP. At subcellular sites with high energy requirements, e.g. at the myofibrillar apparatus during muscle contraction, CK catalyzes the transphosphorylation of PCr to ADP to regenerate ATP, thus preventing a depletion of ATP levels. PCr is thus available as an immediate energy source, serving not only as an energy buffer but also as an energy transport vehicle. Ingestion of creatine increases intramuscular Cr, as well as PCr concentrations, and leads to exercise enhancement, especially in sprint performance. Additional benefits of Cr supplementation have also been noticed for high-intensity long-endurance tasks, e.g. shortening of recovery periods after physical exercise. The present article summarizes recent findings on the influence of Cr supplementation on energy metabolism, and introduces the Cr transporter protein (CreaT), responsible for uptake of Cr into cells, as one of the key-players for the multi-faceted regulation of cellular Cr homeostasis. Furthermore, it is suggested that patients with disturbances in Cr metabolism or with different neuro-muscular diseases may benefit from Cr supplementation as an adjuvant therapy to relieve or delay the onset of symptoms. Although it is still unclear how Cr biosynthesis and transport are regulated in health and disease, so far there are no reports of harmful side effects of Cr loading in humans. However, in this study, we report that chronic Cr supplementation in rats down-regulates in vivo the expression of the CreaT. In addition, we describe the presence of CreaT isoforms most likely generated by alternative splicing. PMID- 9746340 TI - Recommendations for the presentation of NMR structures of proteins and nucleic acids--IUPAC-IUBMB-IUPAB Inter-Union Task Group on the standardization of data bases of protein and nucleic acid structures determined by NMR spectroscopy. AB - The recommendations presented here are designed to support easier communication of NMR data and NMR structures of proteins and nucleic acids through unified nomenclature and reporting standards. Much of this document pertains to the reporting of data in journal articles; however, in the interest of the future development of structural biology, it is desirable that the bulk of the reported information be stored in computer-accessible form and be freely accessible to the scientific community in standardized formats for data exchange. These recommendations stem from an IUPAC-IUBMB-IUPAB inter-union venture with the direct involvement of ICSU and CODATA. The Task Group has reviewed previous formal recommendations and has extended them in the light of more recent developments in the field of biomolecular NMR spectroscopy. Drafts of the recommendations presented here have been examined critically by more than 50 specialists in the field and have gone through two rounds of extensive modification to incorporate suggestions and criticisms. PMID- 9746339 TI - Time-resolved spectroscopy of mitochondria, cells and tissues under normal and pathological conditions. AB - In this study, the detailed dependence of light scattering on tissue architecture and intracellular composition has been investigated. Firstly, we simulated the reduced scattering coefficient (mu(s)') of the rat liver using the Mie theory, the Rayleigh-Debye-Gans approximation and electron microscopy data. Then, the reduced scattering coefficient of isolated rat liver mitochondria, isolated hepatocytes and various rat tissues (i.e. perfused liver, brain, muscle, tumors) was measured at 780 nm by using time-resolved spectroscopy and a sample substitution protocol. The comparison of the isolated mitochondria data with the isolated hepatocyte and whole liver measurements suggests that the mitochondrial compartment is the primary factor for light propagation in hepatic tissue, thus strengthening the relevance of the preliminary theoretical study. Nevertheless, the possibility that other intracellular components, such as peroxisomes and lysosomes, interfere with light propagation in rat liver is discussed. Finally, we demonstrate that light scattering in normal rat tissues and tumors is roughly proportional to the mitochondrial content, according to estimates of the mitochondrial protein content of the tissues. PMID- 9746341 TI - The influence of nickel and cobalt on putative members of the oxygen-sensing pathway of erythropoietin-producing HepG2 cells. AB - Cobalt and nickel stimulate, as does hypoxia, the production of erythropoietin (EPO) in HepG2 cells. Under hypoxic conditions, a decrease in the level of intracellular reactive oxygen species (ROS) is thought to stimulate EPO expression. Cobalt and nickel may interact with the putative oxygen sensor by changing the redox state of the central iron atom of heme proteins, similar to the effects of hypoxia. It was investigated, therefore, whether cobalt and nickel interact with hemeproteins or ROS scavenging systems in the control of intracellular ROS level. Cobalt chloride (100 microM, 24 h) oxidized non respiratory as well respiratory hemeproteins and increased the oxygen consumption. In contrast, nickel chloride (300 microM, 24 h) primarily reduced respiratory hemeproteins and decreased the oxygen consumption. In HepG2 cells treated with CoCl2, iron and cobalt were localized in cytosolic granules close to the cell nucleus and in mitochondria at concentrations up to 12 mM or 41 mM, respectively. Intracellular nickel was not measurable. Three-dimensional reconstruction of confocal laser microscopy images revealed hot spots of hydroxyl radical generation by a Fenton reaction at the sites of cytosolic iron accumulation. The .OH levels decreased in cobalt-treated (to 81%) as well as in nickel-treated (to 67%) HepG2 cells, accompanied by an increase of EPO expression to 167% and 150%, respectively. Our results underline the importance of .OH formed by a Fenton reaction for triggerimg EPO production. Identification of the primary hemeprotein being the oxygen sensor was not possible due to the antagonistic effects of cobalt and nickel on the redox state of detectable hemeproteins. PMID- 9746342 TI - Identification of a gene selectively expressed in the brain, which encodes a putative transmembrane protein and a soluble cytoplasmic isoform. AB - Subtractive cloning procedures led to the identification of a variety of transcripts expressed in mammalian brain. However, little is known about the encoded proteins and the regulation of gene expression. Here, we describe the isolation and characterisation of a single-copy gene (83.5) of 21.7 kb which is specifically expressed in porcine brain. In situ hybridisation and immunohistochemistry experiments showed a distinct pattern of gene expression in neuronal cell types in different parts of the brain. The gene contains two mini exons, confirming neural-specific expression. cDNA cloning experiments revealed two species of mRNA differing in their 5'-regions. These transcripts are generated by two distinct transcription start sites that are under the control of different potential promoter regions as shown by primer-extension experiments. The amino acid sequences of the deduced proteins predict that one mRNA species encodes a novel type-I transmembrane protein, whereas the other transcript encodes only a part of its cytoplasmic domain. In Western-blot experiments, we detected two proteins of the predicted size and cellular localisation in porcine brain. The precise function of these proteins remains to be determined. However, our findings suggest that they may be generated by alternative promoter usage, leading to the expression of a membrane protein and its truncated cytoplasmic isoform. PMID- 9746343 TI - Functional expression of eukaryotic polypeptide chain release factors 1 and 3 by means of baculovirus/insect cells and complex formation between the factors. AB - Translation termination in eukaryotes is governed by termination codons in mRNA and two release factors, eRF1 and eRF3. In this work, human eRF1 and eRF3 have been produced in insect cells using a recombinant baculovirus expression system for the corresponding human cDNAs. Purification of eRF1 has led to a homogeneous 50-kDa protein active in promoting ribosome-dependent and termination-codon dependent hydrolysis of formylmethionyl-tRNAf(Met). Purification of eRF3 yielded a full-length protein and shorter polypeptides. Microsequencing of the N-terminus of the shortest form detected a site of proteolytic cleavage between Arg91 and Gly92, probably due to exposed region(s) hypersensitive to proteolysis. The mixture of full-length and truncated forms of eRF3 as well as bacterially expressed eRF3 lacking 138 N-terminal amino acids (eRF3Cp) are active as an eRF1 dependent and ribosome-dependent GTPase and in stimulating the GTP-dependent release activity of eRF1. Complex formation between eRF1 and eRF3Cp was demonstrated by affinity and gel-filtration chromatographies and by native-gel electrophoresis. An abnormal electrophoretic mobility observed for eRF1 as compared with the complex points to a significant conformational change of either eRF1 or both factors in the complex. Co-expression of both factors in baculovirus infected insect cells and a yeast two-hybrid assay were applied to monitor complex formation in vivo. In yeast cells, both eRF1 and eRF3 are either in a monomeric or in a heterodimeric but not in a homodimeric state. PMID- 9746344 TI - Changes in myosin mRNA and protein expression in denervated rat soleus and tibialis anterior. AB - Denervation differs from other models of reduced neuromuscular activation due to the absence of a nerve-muscle connection and limited data exists regarding the effects of denervation on myosin heavy chain (MHC) expression. Thus, adult MHC expression (I, IIa, IIx, IIb) was studied in the rat soleus and tibialis anterior (TA) at the mRNA and protein levels 2, 4, 7, 10, 14, and 30 days following sciatic nerve transection. MHC protein content was quantified with SDS/PAGE and mRNA levels with the RNase-protection assay. Control soleus consisted predominately of type I MHC mRNA and protein, however, 4 days after denervation type I MHC mRNA was significantly decreased to 41+/-8% of control and continued to remain below control values. Soleus IIa mRNA was significantly elevated 7 and 10 days after denervation while IIx mRNA remained relatively constant until 30 days when it increased to 197+/-23% of control. At the protein level, soleus I MHC significantly decreased to 80% of the total while IIa MHC significantly increased to 20% of the total. At 30 days, Hx MHC protein accounted for 9.4+/ 1.6% of the total soleus MHC protein. In the TA, IIb mRNA was significantly decreased to 57% of control by day 4 and remained significantly decreased for up to a month. TA IIx mRNA was also significantly decreased at 10 and 30 days after denervation. Similar to the soleus, TA Ha mRNA was significantly increased over control 7-14 days after denervation. There were no significant changes in TA MHC protein profile during one month of denervation. In both the soleus and TA, denervation significantly shifted the MHC mRNA profile as early as 4 days following denervation without any corresponding changes at the protein level. Significant mRNA changes without large changes in MHC protein composition continued throughout the denervation period suggesting that the muscle may be prevented from premature functional transitions by mechanisms such as decreased mRNA stability, translational block, or increased turnover of newly synthesized proteins. PMID- 9746345 TI - The transient association of ERp57 with N-glycosylated proteins is regulated by glucose trimming. AB - The thiol-dependent reductase ERp57 has been shown to interact specifically with in vitro synthesised glycoproteins imported into canine pancreatic microsomes. On this basis, it was proposed that ERp57 forms part of a glycoprotein-specific folding 'machinery', present in the lumen of the endoplasmic reticulum (ER). In this study, we have investigated the interaction of ERp57 with newly synthesised proteins using semi-permeabilised mammalian cells (SP cells), in which the ER remains essentially intact and, hence, resembles that of a living cell. We demonstrate that ERp57 interacts preferentially with the glycosylated versions of soluble and membrane proteins, and that this interaction occurs in combination with calnexin and calreticulin. For the first time, we have performed a detailed analysis of the kinetics of ERp57 binding to newly synthesised glycoproteins. We find that ERp57 associates transiently with glycoproteins - a characteristic of molecular chaperones. Using mutant SP cells deficient in glucosidase I, we confirm that the binding of ERp57 to glycoproteins depends upon glucose trimming. We also demonstrate, for the first time, that the release of ERp57 from glycoprotein substrates is dependent upon glucose trimming. These data are combined to present a unified model for the role of ERp57/ER lectin complexes during glycoprotein folding in vivo. PMID- 9746346 TI - An adrenocorticotropin-regulated phosphoprotein intermediary in steroid synthesis is similar to an acyl-CoA thioesterase enzyme. AB - We have previously reported the purification of a phosphoprotein (p43) intermediary in steroid synthesis from adrenal zona fasciculata [Paz C., Dada, L. A., Cornejo Maciel, M. F., Mele, P. G., Cymeryng, C. B., Neuman, I., Mendez, C. F., Finkielstein, C. V., Solano, A. R., Park, M., Fischer, W. H., Towbin, H., Scartazzini, R. & Podesta, E. J. (1994) Eur J. Biochem. 224, 709-716]. Here, we describe the cloning and sequencing of a cDNA encoding p43 as well as the hormonal regulation of the p43 transcript. The protein resulted homologous to a very recently described mitochondrial peroxisome-proliferator-induced very-long chain acyl-CoA thioesterase (MTE-I). The deduced amino acid sequence of the protein shows consensus sites for phosphorylation by different protein kinases, and a lipase serine motif. Antibodies raised against a synthetic peptide that includes the lipase serine motif and against the N-terminal region of p43 block the action of the protein. The transcript of p43 was detected in ovary of pseudopregnant rats, rat adrenal zona fasciculata and glomerulosa, mouse Leydig tumor cell line (MA-10), rat brain and human placenta. Inhibition of adrenocorticotropin hormone (ACTH) release and steroid synthesis by dexamethasone produced a dose-dependent decrease in the abundance of the adrenal transcript. The transcript was induced by in vivo stimulation of the adrenals with ACTH. The effect had a rapid onset (5 min), reached maximal stimulation (62%) at 15 min, and returned to basal levels at 30 min. The effect of ACTH on the p43 transcript was inhibited by actinomycin D and enhanced by cycloheximide. Our results provide the first evidence linking acyl-CoA thioesterases with very-long-chain specificities, and a protein intermediary in steroid synthesis, thereby supporting a regulatory role for acyl-CoA thioesterases in steroidogenic tissues. PMID- 9746348 TI - Direct effects of corticotrophin on oral keratinocyte cell proliferation. AB - Corticotropin is produced by keratinocytes and may have an immunoregulatory role in oral mucosa and skin. We have investigated its effects on a human oral keratinocyte cell line and shown that corticotropin, acting via its specific receptor, stimulates a dose-dependent increase in DNA synthesis and induces cell proliferation. When cells were incubated in the presence of increasing concentrations of corticotropin, there were significant increases in intracellular cAMP levels. Corticotropin-stimulated mitogenesis and cell proliferation were attenuated by the adenylyl cyclase inhibitor SQ22,536, but were unaffected by inhibitors of protein kinase C or tyrosine kinase. These data identify corticotropin as a mitogenic regulatory peptide of keratinocytes acting via cAMP. PMID- 9746347 TI - Characterization of promoter regions involved in high expression of KlCYC1. AB - Functional analysis of the KlCYC1 promoter reveals that sequences located upstream to those already published [Freire-Picos, M. A., Rodriguez-Torres, A. M., Ramil, E., Cerdan, M. E., Breuning, K. D., Hollenberg, C. P. & Zitomer, R. S. (1993) Sequence of a cytochrome c from Kluyveromyces lactis and its upstream region, Yeast 9, 201-204] and extending from positions -780 to -371 are important for maintaining high levels of expression, although this region contains both negative and positive elements. A consensus sequence for interaction with KlCpf1p is present at position -492, into the negative site, and specific protein binding to KlCpf1p has been demonstrated. Deletion of the sequences from positions -413 to -338 diminishes KlCYC1 transcription; protein binding to two sequences included in this activator region is detected and several points of evidence indicate that the complex observed is different from the Hap2/3/4/5p complex. Binding of KlCpf1p and the activator complex to the promoter is constitutive in different carbon sources. Although the promoter contains CCAAT boxes, directed mutagenesis has revealed that they are not related to the moderate de-repression observed in glycerol media. PMID- 9746349 TI - Modular evolution of the Glx-tRNA synthetase family--rooting of the evolutionary tree between the bacteria and archaea/eukarya branches. AB - The accuracy of protein biosynthesis generally rests on a family of 20 aminoacyl tRNA synthetases, one for each amino acid. In bacteria, archaea and eukaryotic organelles, the formation of Gln-tRNA(Gln) is prevalently accomplished by a transamidation pathway, aminoacylation of tRNA(Gln) with Glu by glutamyl-tRNA synthetase (GluRS) followed by a tRNA-dependent transamidation of Glu from Glu tRNA(Gln). A few bacterial species, such as Escherichia coli, possess a glutaminyl-tRNA synthetase (GlnRS), responsible for Gln-tRNA(Gln) formation. Phylogenetic analysis of the GluRS or GlnRS families (GlxRS) suggested that GlnRS has a eukaryotic origin and was horizontally transferred to a restricted set of bacteria. We have now isolated an additional GlnRS gene from the plant Lupinus luteus and analyzed in more details the modular architecture of the paralogous enzymes GluRS and GlnRS, starting from a large data set of 33 GlxRS sequences. Our analysis suggests that the ancestral GluRS-like enzyme was solely composed of the catalytic domain bearing the class-defining motifs of aminoacyl-tRNA synthetases, and that the anticodon-binding domain of GlxRSs was independently acquired in the bacteria and archaea branches of the universal tree of life, the eukarya sub-branch arising as a sister group of archaea. The transient capture of UAA and UAG codons could have favored the emergence of a GlnRS in early eukaryotes. PMID- 9746350 TI - Two families of sterol methyltransferases are involved in the first and the second methylation steps of plant sterol biosynthesis. AB - Two methyl transfers are involved in the biosynthesis of 24-methyl and 24-ethyl sterols, which play major roles in plant growth and development. The first methyl transfer applies to cycloartenol, the second to 24-methylene lophenol. About ten cDNA clones encoding S-adenosyl-L-methionine (AdoMet) sterol methyltransferases (SMTs) have been isolated so far from various plants. According to their deduced amino acid sequences, they were classified in two families, smtl and smt2; in addition, smt2 cDNAs were shown to encode a 24-methylene lophenol C24 methyltransferase [Bouvier-Nave, P., Husselstein, T., Desprez, T. & Benveniste, P. (1997) Eur. J. Biochem. 246, 518-529]. We now report the comparison of two cDNAs isolated from Nicotiana tabacum, Ntsmt1-1 which belongs to the first SMT cDNA family and Ntsmt2-1 which belongs to the second. Both cDNAs were expressed in the yeast null mutant erg6, deficient in SMT. Whereas erg6 is devoid of 24 alkyl sterols, erg6 Ntsmt1-1 contained a majority of 24-methylene sterols and erg6 Ntsmt2-1, a majority of 24-ethylidene sterols, indicating distinct functions for the expression products of these cDNAs. In the presence of AdoMet, delipidated microsomes from erg6 Ntsm1-1 efficiently converted cycloartenol into 24-methylene cycloartanol, but did not produce any 24-ethylidene lophenol upon incubation with 24-methylene lophenol. This demonstrates that cDNA Ntsmt1-1 (and most probably the other plant SMT cDNAs of the first family) encode(s) a cycloartenol C24 methyltransferase. In contrast, delipidated microsomes of erg6 Ntsmt2-1 were shown to methylate preferentially 24-methylene lophenol, as expected from an SMT encoded by an smt2 cDNA. In summary, among various cDNAs isolated from N. tabacum, one (Ntsmt1-1) belongs to the first family of plant SMT cDNAs according to its deduced amino acid sequence and was shown to encode a cycloartenol C24 methyltransferase, whereas another (Ntsmt2-1) belongs to the second family and was shown to encode a 24-methylene lophenol C24 methyltransferase. Meanwhile, two cDNAs were isolated from Oriza sativa and shown to belong to smtl and to smt2 families, respectively. These data disclose the coexistence, in a given plant species, of two distinct SMTs, each catalyzing one step of methylation in the sterol biosynthesis pathway. PMID- 9746351 TI - Interaction of ribosomal L1 proteins from mesophilic and thermophilic Archaea and Bacteria with specific L1-binding sites on 23S rRNA and mRNA. AB - In Bacteria and Archaea (formerly Archaebacteria) ribosomal protein L1 has a dual function, as a primary rRNA-binding protein and as a translational repressor which binds to its own mRNA. The L1-binding site on the mRNA exhibits high similarity in both sequence and secondary structure to the binding site for L1 on the 23 S rRNA. A sensitive membrane-filter-binding assay has been used to examine the interactions between ribosomal L1 proteins from different archaeal and bacterial species, and 23S rRNA and mRNA fragments from Methanococcus vannielii containing the MvaL1-binding site. Under standard conditions (0 degrees C, pH 7.5, 20 mM Mg2+, 500 mM KCl), the apparent dissociation constant Kd of the homologous MvaL1-23S rRNA complex is 5 nM, the apparent dissociation constant Kd of the MvaL1-mRNA complex is 0.15 degrees M. L1 proteins from Escherichia coli (EcoL1) and from the thermophilic Bacterium Thermus thermophilus (TthL1), and from the thermophilic Archaea Methanococcus thermolithotrophicus (MthL1), Methanococcus jannaschii (MjaL1), and Sulfolobus solfataricus (SsoL1) were tested for their affinity to the specific L1-binding sites on the 23 S rRNA and mRNA. In general, the affinity of L1 proteins from thermophilic species to the binding sites on both 23 S rRNA and mRNA is about one order of magnitude higher than that of their mesophilic counterparts. This stronger protein-RNA interaction might make a substantial contribution to the thermal tolerance of ribosomes in thermophilic organisms. PMID- 9746353 TI - Purification, characterization and complete amino acid sequence of nuclease C1 from Cunninghamella echinulata var. echinulata. AB - It is known, from the zymogram method of nuclease activity assay, that the crude extracts of Cunninghamella echinulata var. echinulata contained at least three distinct extracellular nucleases. Among them, the major form was 30 kDa in molecular mass and termed nuclease C1. In this report, nuclease C1 was purified to apparent homogeneity by chromatography on Cibacron blue-3GA, phenyl-Sepharose 4B and HiTrap Heparin. Nuclease C1 acquired enzymatic activity in the presence of Mn2+ or Mg2+ and was inhibited by EDTA. The activity was maximal at pH 7-8.5. The primary structure of nuclease C1 was completely determined using enzymatic digestion and gene cloning. The N-terminal 49 residues of nuclease C1 were first elucidated from a tryptic digest. Two degenerate upstream primers were subsequently designed to amplify the cDNA encoding nuclease C1. The resulting protein sequence of nuclease C1 was shown to be composed of 252 residues. It was intriguing to find that the protein sequence of nuclease C1 showed significant similarities with the sequences of the mitochondrial nucleases of Saccharomyces cerevisiae (44% identity) and Schizosaccharomyces pombe (42% identity). Residue His87 of nuclease C1 was postulated to be located at the active site from sequence similarity with secreted nuclease from Serratia marcescens. PMID- 9746352 TI - The bZip transcription factor vitellogenin-binding protein is post transcriptional down regulated in chicken liver. AB - The vitellogenin-binding protein (VBP) is a member of the proline and acidic region rich (PAR) family of bZip transcription factors. PAR is located N terminally to the DNA-binding domain. VBP binds to specific sites within the 300 bp 5'-flanking region of the chicken-liver-specific estrogen-dependent very-low density apolipoprotein gene (apoVLDL II). One of these binding sites (site D) resembles the albumin site D and is positioned in close proximity of the major estrogen-responsive element. Previous studies showed that VBP can bind simultaneously with the estrogen receptor to the putative complex regulatory element E1D. To investigate whether VBP is involved in apoVLDL II gene expression, we examined its capacity to enhance apoVLDL II transcription and its presence in liver. We show that VBP is capable of enhancing transcription in transfection experiments. However, VBP could not be detected in liver by Western blots or immuno-electro mobility shift assays (EMSAs) using antibodies against different moieties of the protein. We examined the possible reduction in translation efficiencies due to a small upstream open reading frame in the VBP leader sequence, but did not find any. Although VBP binds to the proximal apoVLDL II promoter region and enhances transcription in co-transfection experiments, the protein is unlikely to be involved in apoVLDL II gene transcription because of its undetectable low level in liver nuclei. PMID- 9746354 TI - Modified glycosylation of cellobiohydrolase I from a high cellulase-producing mutant strain of Trichoderma reesei. AB - Cellobiohydrolase I is an industrially important exocellulase secreted in high yields by the filamentous fungus Trichoderma reesei. The nature and effect of glycosylation of CBHI and other cellulolytic enzymes is largely unknown, although many other structural and mechanistic aspects of cellulolytic enzymes are well characterised. Using a combination of liquid chromatography, electrospray mass spectrometry, solid-phase Edman degradation, and monosaccharide analysis we have identified every site of glycosylation of CBHI from a high cellulase-producing mutant strain of T. reesei, ALKO2877, and characterised each site in terms of its modifying carbohydrate and site-specific heterogeneity. The catalytic core domain comprises three N-linked glycans which each consist of a single N acetylglucosamine residue. Within the glycopeptide linker domain, all eight threonines are variably glycosylated with between at least one, and up to three, mannose residues per site. All serines in this domain are at least partially glycosylated with a single mannose residue. This linker region has also been shown to be sulfated by a combination of ion chromatography and collision-induced dissociation electrospray mass spectrometry. The sulfate is probably mannose linked. The biological significance of N-linked single N-acetylglucosamine in the catalytic core, and mannose sulfation in the linker region, is not known. PMID- 9746355 TI - Interactions of non-detergent sulfobetaines with early folding intermediates facilitate in vitro protein renaturation. AB - Non-detergent sulfobetaines (NDSB) are a family of solubilizing and stabilizing agents for proteins. In a previous study [Goldberg, M. E., Expert-Bezancon, N., Vuillard, L. & Rabilloud, T. (1996) Folding & Design 1, 21-27] we showed that the amount of active protein recovered in in vitro folding experiments could be significantly increased by some NDSBS. In this work we investigated the mechanisms by which these molecules facilitate protein renaturation. Stopped-flow and manual-mixing fluorescence and enzyme activity measurements were used to compare the kinetics of protein folding in the presence and absence of N-phenyl methyl-N,N-dimethylammonium-propane-sulfonate (NDSB 256). Hen lysozyme and the beta2 subunit of Escherichia coli tryptophan synthase were chosen as model systems since their folding pathways had been previously investigated in detail. It is shown that, massive aggregation of tryptophan synthase occurs within less than 2.5 s after dilution in the renaturation buffer, but can be prevented by NDSB 256; only very early folding phases (such as the formation of a loosely packed hydrophobic core able to bind 8-anilino-1-naphthalenesulphonic acid, and the initial burying of tryptophan 177) are significantly altered by NDSB 256; none of the later phases is affected. Furthermore, NDSB 256 did not significantly affect any of the kinetic phases observed during the refolding of denatured lysozyme retaining intact disulphide bonds. This shows that NDSB 256 only interferes with very early steps in the folding process and acts by limiting the abortive interactions that could lead to the formation of inactive aggregates. PMID- 9746356 TI - Studies on the Zn-containing S14 ribosomal protein from Thermus thermophilus. AB - The S14 ribosomal protein from the thermophilic organism Thermus thermophilus, which contains a zinc-finger-like motif, namely -C-X2-C-X12-C-X2-C- [Tsiboli, P. & Choli, T. (1995) Biol. Chem. Hoppe-Seyler 376, 127-130], has been overproduced, purified and investigated for its zinc content. According to atomic absorption experiments, the protein contains zinc at a molar ratio of one. Denaturation experiments with simultaneous use of denaturing and chelating agents (guanidine hydrochloride and EDTA), as well as renaturation experiments, have shown both that Zn is strongly bound to the protein and with 1:1 Zn/protein stoicheiometry. These findings provide very strong evidence in support of the participation of the zinc-finger motif and the Zn in the formation of a zinc-finger domain. PMID- 9746357 TI - Interactions of elongation factor 2 with the cytoskeleton and interference with DNase I binding to actin. AB - Interactions of elongation factor 2 (EF-2) with G-actin and F-actin in vitro were investigated using viscosimetry, gel filtration and electron microscopy. Under depolymerization conditions, at a molar ratio of 0.5:1 (EF-2/F-actin subunit), F actin is stabilised by EF-2 and filaments depolymerize about three times slower than control solutions containing only F-actin. Filament stability is improved also when EF-2 is included in the solution in the presence of DNase I. Electron micrographs and viscosity measurements indicate that EF-2 may support small bundles with a width of 2 or 3 filaments. It was established that EF-2 interacts with G-actin in vitro, and reduces G-actin inhibition of DNase I activity when it is present at a ratio of 1:1. Results are discussed in the context of possible functional significance of the interactions. PMID- 9746358 TI - Genes coding for the benzoyl-CoA pathway of anaerobic aromatic metabolism in the bacterium Thauera aromatica. AB - Many aromatic compounds are anaerobically oxidized to CO2 via benzoyl-CoA as the common aromatic intermediate. In Thauera aromatica, the central benzoyl-CoA pathway comprises the ATP-driven two-electron reduction of the benzene ring; this reaction uses a ferredoxin as electron donor and is catalyzed by benzoyl-CoA reductase. The first intermediate, cyclohex-1,5-diene-1-carboxyl-CoA, is subsequently hydrated by dienoyl-CoA hydratase to 6-hydroxycyclohex-1-ene-1 carboxyl-CoA. Formation of the main product produced by cell extracts, 3 hydroxypimelyl-CoA, requires at least two further steps; the oxidation of a hydroxyl group and the hydrolytic carbon ring cleavage of a CoA-activated beta oxoacid. In addition, enoyl-CoA hydratase may come into play. A cluster of eight adjacent genes, which are transcribed in the same direction and may form an operon, was found in this bacterium. The cluster codes for proven and postulated enzymes of the benzoyl-CoA pathway. The genes for the enzymes code for ferredoxin, four subunits of benzoyl-CoA reductase, dienoyl-CoA hydratase, 6 hydroxycyclohex-1-ene-1-carboxyl-CoA dehydrogenase (NAD+), and the ring hydrolyzing enzyme. The deduced amino acid sequences of these proteins were 35 86% similar to the corresponding sequences found in Rhodopseudomonas palustris. Benzoyl-CoA reductase subunits exhibit distinct similarities with 2 hydroxyglutaryl-CoA dehydratase and its ATP-hydrolysing activase protein of Acidaminococcus fermentans as well as with open reading frames of unknown function in other bacteria. Conversion of benzoyl-CoA to 3-hydroxypimelyl-CoA can be explained by a minimal model of the benzoyl-CoA pathway assuming the four enzymes whose genes were characterized and an additional enoyl-CoA hydratase. In R. palustris the dienoyl-CoA hydratase gene is lacking suggesting the operation of a modified benzoyl-CoA pathway with cyclohex-1-ene-1-carboxyl-CoA as intermediate. PMID- 9746359 TI - The catalytic mechanism of adenosylhomocysteine/methylthioadenosine nucleosidase from Escherichia coli--chemical evidence for a transition state with a substantial oxocarbenium character. AB - The substrate and inhibitory specificity of Escherichia coli adenosylhomocysteine (AdoHcy)/methylthioadenosine (MeSAdo) nucleosidase has been explored with several MeSAdo analogues modified on the sugar moiety at the 2', 3' and 5' positions. Alteration at C3' or at C2' and C3' positions in MeSAdo abolished substrate activity. However, the 2'-deoxy analogue of MeSAdo is effective as a substrate; this result provides evidence against a possible general-base catalysis involving the anchimeric assistance of the 2'-alpha-hydroxy group and the formation of a 1,2-epoxide as an intermediate in the catalytic process. The results of a study of the interaction of an 8,5'-cyclo analogue of MeSAdo with the enzyme indicate the importance of the glycosidic conformation of the substrate for binding to the active site. The enzyme discriminates against methanol attack from the solvent during catalysis. This implies the participation of an enzyme-directed water nucleophile. A poor solvent kinetic deuterium-isotope effect was measured (0.93) on the Vmax. Plots of log Vmax and log (Vmax/Km) for MeSAdo as a function of pH values from 5.0 to 8.5 are similar, with two presumably essential ionisable groups for catalysis with apparent pKa values of 5.6 and 8.2, whereas Km is independent of pH. When the 2'-alpha-hydroxy group of MeSAdo is substituted by fluorine, a significant decrease (28 500-fold) in the Vmax for enzyme-catalysed hydrolysis of the modified substrate is observed. This result indicates a transition state with a substantial oxocarbenium character. From these data, the reaction mechanism for AdoHcy/MeSAdo nucleosidase is discussed. PMID- 9746360 TI - The structures of asparagine-linked oligosaccharides of rat liver cathepsin L reflect the substrate specificity of lysosomal alpha-mannosidase. AB - We have studied the structures of asparagine-linked oligosaccharides of cathepsin L purified from rat liver in detail. The oligosaccharides released from rat liver cathepsin L on glycopeptidase-F treatment were tagged with 2-aminopyridine at their reducing ends. The pyridylamino (PA) derivatives were separated into seven fractions according to molecular size by normal-phase HPLC. The structure of each oligosaccharide thus isolated was analyzed by reversed-phase HPLC and characterized by ion-spray mass spectrometry and high-resolution proton nuclear magnetic resonance (1H-NMR) spectroscopy. Our results indicate that the asparagine-linked oligosaccharides of rat liver cathepsin L are of the oligomannose type, having two to six mannose residues. Among them, the five major ones are Manalpha1-6Manbeta1-4-GlcNAcbeta1-4GlcNAc, Manalpha1 -6Manalpha1 6Manbeta1-4GIcNAcbeta1-4GlcNAc, Manalpha1-6(Manalpha1-3)-Manalpha1-6Manbeta1- 4GlcNAcbeta1-4GlcNAc, Manalpha1-6(Manalpha1-3)Manalpha1-6(Manalpha1-3) Manbeta1 4Glc-NAcbeta1-4GlcNAc, and Manalpha1-6(Manalpha1-3)Manalpha1-6(Manalpha1-++ +2Manalpha1-3)Manbeta1-4GlcNAcbeta1-4Glc-NAc. Their structures are shown to be products of Man6GlcNAc2 hydrolysis with lysosomal alpha-mannosidase. PMID- 9746361 TI - The ATPase and ATP-binding functions of P-glycoprotein--modulation by interaction with defined phospholipids. AB - P-glycoprotein functions as an active efflux pump for lipophilic compounds and plays an important role in the resistance of human cancers to chemotherapeutic drugs. Drug transport is powered by ATP hydrolysis at two highly conserved nucleotide-binding domains, which are proposed to be located at the cytosolic face of the protein. The ATPase activity of P-glycoprotein depends on the presence of phospholipids, and various lipids affect both basal ATPase activity and its stimulation or inhibition by drug substrates. The modulating effects of the lipid-phase state and effects on the function of the nucleotide-binding domains of P-glycoprotein have been studied in reconstituted vesicles of the synthetic phospholipids 1-palmitoyl-2-myristoylphosphatidylcholine (PamMyrGroPCho) and dimyristoylphosphatidylcholine (Myr2GroPCho). The kinetic parameters for P-glycoprotein ATPase activity were determined, and a fluorescence quenching technique was used to measure the Kd for ATP binding. The values of both the Km for ATP hydrolysis and Kd for ATP binding were significantly different above and below the gel/liquid-crystalline phase transition temperature (tm) of PamMyrGroPCho and Myr2GroPCho, whereas they were similar at the same temperatures for P-glycoprotein in detergent solution. A discontinuity at 21-24 degrees C was observed in the Arrhenius plots of P-glycoprotein ATPase activity in a membrane environment, but not in detergent solution. In addition, the activation energies for ATP hydrolysis in the gel and liquid-crystalline phases of the lipid bilayer were significantly different. P-glycoprotein in PamMyrGroPCho bilayers displayed an unusually low activation energy just below the melting transition. These results indicate that both ATP binding and ATP hydrolysis by P-glycoprotein are affected by the phase state of the host lipids in which it is reconstituted. Lipids may modulate the function of the nucleotide binding domains of P-glycoprotein by interacting with the transmembrane regions of the protein, or the nucleotide-binding domains themselves may interact with the surface of the bilayer. PMID- 9746363 TI - Pro-oxidant role of superoxide dismutase in ultraviolet-A-induced lipid peroxidation in cultured normal human skin fibroblasts. AB - Exposure of cultured normal human skin fibroblasts to ultraviolet A triggers lipid peroxidation. In sharp contrast with the tert-butylhydroperoxide-induced lipid peroxidation, the ultraviolet-A-induced lipid peroxidation is inhibited by treating cells with diethyldithiocarbamate. Diethyldithiocarbamate decreases superoxide dismutase activity and, to a lesser extent, the total glutathione level. Catalase and glutathione peroxidase, however, are unaffected. The decrease in the superoxide dismutase activity parallels an inhibition of H2O2 formation in both irradiated and unirradiated cells. The protection against lipid peroxidation may thus be associated with superoxide dismutase inhibition. Membrane damage revealed by neutral red uptake is not prevented by diethyldithiocarbamate. PMID- 9746362 TI - Use of a photoactivatable lipid to probe the topology of PA63 of Bacillus anthracis in lipid membranes. AB - The protective antigen of Bacillus anthracis is a key protein that promotes the translocation of the enzymatic moieties of the two toxins of B. anthracis into the cell cytoplasm. The membrane topology of the active form of the protective antigen (PA63) was investigated by proteolysis of PA63 inserted into liposomes containing a photoactivatable, radioactive lipid, and characterization of the N terminal moiety of the deeply-inserted (and therefore radiolabeled) peptides. A single sequence starting at residue Ala258 was identified. Fourier-transform infrared spectroscopy showed that the protected peptide was mainly adopting a beta-sheet structure whose orientation was compatible with a transmembrane organization. PMID- 9746364 TI - Adrenocorticotropic hormone stimulates CYP11B1 gene transcription through a mechanism involving AP-1 factors. AB - To elucidate the mechanism(s) by which adrenocorticotropic hormone (corticotropin) stimulates transcription of the steroid 11beta-monooxygenase gene (CYP11B1) in adrenocortical cells, the 5'-flanking region of rat CYP11B1 was analyzed using transient transfection and protein-binding assays with mouse adrenocortical Y1 cells. The results indicated that both basal and corticotropin induced transcriptional activation of CYP11B1 required a common regulatory element containing a binding site for activator protein-1 (AP-1) transcription factors (dimers of the Jun and Fos family proteins) in the 5'-flanking region. Other DNA-binding protein(s) such as transcription factor Ad4BP was not required for either basal or corticotropin-induced transcriptional activation. Corticotropin stimuli were found to induce expression of a subset of the jun and fos family gene products in Y1 cells significantly, while total amounts of AP-1 factors capable of binding to its site in the CYP11B1 promoter did not change greatly. Treatment of rats with corticotropin had similar effects on mRNA levels of the jun and fos family genes in the adrenocortical zona fasciculata cells together with an enhancing effect on the level of CYP11B1 mRNA in the tissue. The effects of corticotropin on mRNA levels of the jun and fos family genes as well as transcription of CYP11B1 in Y1 cells were mimicked by treatment of the cells with dibutyryl cAMP. Furthermore, when components of AP-1 factors were overexpressed by transfecting Y1 cells with their expression vectors, a paired expression of AP-1 components such as c-Jun and c-Fos, which were inducible by corticotropin, transactivated the CYP11B1 promoter more strongly in the absence of corticotropin than other combinations such as JunD and Fra-2 expressed constitutively. These results suggest that corticotropin regulates transcription of the CYP11B1 gene by causing compositional changes in AP-1 transcription factors in the adrenocortical cells via a cAMP-dependent pathway. PMID- 9746366 TI - A regulatory factor, Fil1p, involved in derepression of the isocitrate lyase gene in Saccharomyces cerevisiae--a possible mitochondrial protein necessary for protein synthesis in mitochondria. AB - A mutant was isolated that failed to derepress the 5' upstream region of Candida tropicalis isocitrate lyase gene (UPR-ICL)-mediated gene expression in acetate medium, and the gene (FIL1) that complemented this mutation was isolated. The fil1 null mutant in which FIL1 is disrupted (deltafil1 strain) could not grow on acetate or ethanol, and the derepression of the isocitrate lyase encoded by ICL1 in Saccharomyces cerevisiae was also defected. The amino acid sequence of Fil1p (230 amino acids) showed similarity to ribosome recycling factors (RRFs) of prokaryotes. Compared to prokaryotic RRFs, Fil1p had an N-terminal 46-amino-acid extension which was shown to be able to function as a mitochondrial-targeting sequence. The subcellular fractionation of the deltafil1 strain showed that protein constituents of the mitochondrial fraction of the deltafil1 strain differed from those of the wild-type strain, but resembled those of chloramphenicol-treated cells or rho(o) cells. The specific activity of cytochrome c oxidase, was severely decreased in deltafil1, rho(o) and chloramphenicol-treated cells compared with wild-type cells, while enzymatic levels of mitochondrial NAD+-linked isocitrate dehydrogenase, which is encoded by nuclear DNA, were not affected. These results suggest that Fillp is necessary for protein synthesis in mitochondria of S. cerevisiae. Furthermore, cells treated with antimycin A, along with chloramphenicol-treated, rho(o), and deltafil1 cells, showed deficiency in derepression of isocitrate lyase. Northern-blot analysis showed that this can be ascribed to no increase in transcription of ICL1 and FBP1 encoding fructose 1,6-bisphosphatase. The results indicate the presence of a communication pathway between mitochondria and the nucleus which represses expression of genes encoding the key enzymes of the glyoxylate cycle and gluconeogenic pathway when there is a deficiency in the mitochondrial respiratory chain. PMID- 9746365 TI - PACSIN, a brain protein that is upregulated upon differentiation into neuronal cells. AB - To identify genes that are differentially expressed during self-repair processes in mouse brain, we screened a subtracted cDNA library enriched for brain-specific clones. One of these clones, H74, detected a 4.4-kb mRNA predominantly expressed in brain and dorsal root ganglia neurons. Expression increased continuously during the lifespan and the state of differentiation, but decreased after entorhinal-cortex lesion. A full-length cDNA clone was isolated from a cerebellum cDNA library and characterized. Sequence analysis and database search revealed high sequence similarity to FAP52, a protein expressed in focal-adhesion contacts, and uncharacterized Echinococcus and Caenorhabditis elegans gene products. Furthermore, peptide sequences derived from human cDNA fragments showed up to 65% sequence identity at the amino acid level. The presence of a C-terminal src homology 3 (SH3) domain and its phosphorylation by casein kinase 2 (CK2) and protein kinase C (PKC) imply a role in signaling. Here we demonstrate that the gene encodes a phosphoprotein, referred to as PACSIN, with a restricted spatial and temporal expression pattern. PMID- 9746367 TI - Preferential pre-mRNA utilisation of an upstream cryptic 5' splice site created by a single base deletion mutation in exon 37 of the FBN-1 gene. AB - A heterozygous deletion of a single base (A4704) from exon 37 of the fibrillin-1 gene was defined in a patient with Marfan syndrome and subsequently in his previously undiagnosed father. The deletion created a cryptic 5' splice site in exon 37 which was utilised in preference to the normal 5' splice site during pre mRNA processing in skin fibroblasts cultured from the proband. The mutant mRNA showed a 48-bp deletion from the 3' end of exon 37 which was predicted to restore the reading frame in the mutant mRNA and result in the deletion of a 16-amino acid sequence from a central eight-cysteine repeat motif of the fibrillin-1 molecule. Interestingly, the cryptic 5' splice site in exon 37 and the normal 5' splice site had equally strong consensuses for splice-site selection. The preferential utilisation of the cryptic site is discussed in relation to current theories on the mechanisms involved in pre-mRNA splicing. Analysis by reverse transcription PCR indicated that, in the patients skin fibroblasts, the steady state level of the mis-spliced mutant mRNA was close to that from the normal allele. In addition, evidence from immunoblotting and pulse-chase biosynthetic labelling indicated that close to normal amounts of fibrillin-1 were being synthesised and secreted by the cells. However, in contrast to control cells cultured from an unaffected individual, little fibrillin-1 was detected, either biosynthetically or by immunofluorescence, in the extracellular matrix produced by the proband's fibroblasts. Thus, the slightly shorter mutant fibrillin-1 molecules appeared to be exerting a powerful dominant-negative effect on the incorporation of normal fibrillin-1 molecules into microfibrils in this culture system. This severe inhibition of microfibril synthesis in cell culture contrasts with the 'classic' phenotype of the proband, suggesting that factors influencing microfibril formation may differ greatly between in vivo and in vitro environments. PMID- 9746368 TI - Characterization of the interaction of myosin with ATP analogues having the syn conformation with respect to the adenine-ribose bond. AB - Numerous analytical experiments have shown that, in solution, ATP analogues with bulky substitutions at the eighth position of the adenine ring predominantly assume the syn conformation with respect to the adenine-ribose bond. Two such analogues, 3'-O-(N-methylanthraniloyl)-8-azido-ATP (Mant-8-N3-ATP) and 8-Br-ATP, were synthesized and used to probe the conformation of the ATP-binding site of myosin. In the presence of these analogues, actomyosin was rapidly dissociated; Mg2+-dependent ATP hydrolysis was significantly activated by actin; and Pi bursting was observed. For skeletal myosin, however, these analogues failed to support actin translocation, and they did not significantly enhance the intrinsic tryptophan fluorescence of skeletal muscle myosin subfragment-1 (SKE S-1). These results suggest that although myosin**/ADP/Pi intermediates can be formed with these analogues, the crucial conformational changes required for cross-bridge cycling do not occur in skeletal muscle myosin. The conformations of the ATP binding sites of skeletal and smooth-muscle myosin were compared using the ternary complexes, myosin-ADP-beryllium fluoride (BeFn) or myosin-ADP-aluminium fluoride (AIF4-). In AlF4- complexes, Mant-8-N3-ADP affinity labeled the N terminal 29-kDa domain of smooth-muscle myosin subfragment-1 (SM S-1), as did ATP analogues having the anti conformation, whereas it labeled the C-terminal 20-kDa domain of skeletal S-1. In smooth muscle BeFn complexes, Mant-8-N3-ADP was equally likely to cross-link to the 29-kDa N-terminal and the 25-kDa C-terminal domains. These analogues induced smooth muscle actomyosin super-precipitation and increased intrinsic tryptophan fluorescence to the same degree as ATP itself. As was expected from above results, the analogues supported smooth-muscle-myosin induced actin translocation. These results suggest that smooth-muscle myosin adopts the eight-substituted ATP analogue in the normal conformation, but skeletal muscle myosin does not. This reflects the likely differences in the structures of their respective ATPase sites. PMID- 9746369 TI - Beta-amyrin synthase--cloning of oxidosqualene cyclase that catalyzes the formation of the most popular triterpene among higher plants. AB - Beta-amyrin, a typical pentacyclic triterpene having an oleanane skeleton, is one of the most commonly occuring triterpenes in nature and is biosynthesized from (3S)-2,3-oxidosqualene. The enzyme, beta-amyrin synthase, catalyzing the cyclization of oxidosqualene into beta-amyrin, generates five rings and eight asymmetric centers in a single transformation. A homology-based PCR method was attempted to obtain the cDNA of this enzyme from the hairy root of Panax ginseng which produces oleanane saponins together with dammarane-type saponins. Two sets of degenerate oligonucleotide primers were designed at the regions which are highly conserved among known oxidosqualene cyclases (OSCs). Nested PCRs using these primers successfully amplified the core fragment which revealed the presence of two OSC clones PNX and PNY. Specific amplification of each clone by 3'-RACE and 5'-RACE was carried out to obtain the whole sequences. The two clones exhibited 60% amino acid identity to each other. A full-length clone of PNY was ligated into the yeast expression vector pYES2 under the GAL1 promoter to give pOSC(PNY). Beta-amyrin production was observed with the mutant yeast lacking lanosterol synthase, transformed by this plasmid. The sequence of pOSC(PNY) contains an open reading frame of 2289 nucleotides which codes for 763 amino acids with a predicted molecular mass of 88 kDa. Sequence comparison with other OSCs showed a high level of similarity with lanosterol, cycloartenol and lupeol synthases. The other clone, pOSC(PNX), was shown to be cycloartenol synthase by similar expression in yeast. The present studies have revealed that distinct OSC exists for triterpene formation in higher plants, and the high level of similarity with cycloartenol synthase indicates close evolutional relationship between sterol and triterpene biosynthesis. PMID- 9746370 TI - Characterization of glycerol uptake in bloodstream and procyclic forms of Trypanosoma brucei. AB - Uptake of glycerol was studied in bloodstream and insect forms of the African parasite Trypanosoma brucei using [14C]glycerol in combination with the oil centrifugation technique. Our kinetic measurements revealed that in bloodstream forms glycerol appeared to be transported by two different mechanisms: firstly by a facilitated-diffusion carrier showing a Km of 0.17 mM and a Vmax of 44 nmol 10( 8) cells min(-1) that predominates at low glycerol concentrations, and secondly by simple diffusion. The effects induced by various inhibitors suggest that uptake is neither sodium dependent nor proton-motive-force driven. The saturable component of transport was phloretin and cytochalasin B sensitive and could also be inhibited by the substrate analogue glyceraldehyde, which led to a 74% decrease in glycerol uptake. In insect forms, however, glycerol is taken up by simple diffusion only. Uptake was insensitive to mercury ions and was not influenced by a variety of different channel inhibitors. Our data show that in T brucei glycerol transport across the plasma membrane occurs by simple diffusion. In addition, bloodstream forms express a carrier protein which promotes a rapid transport at low glycerol concentrations. Expression of this transport protein may account for a selective secretion of intracellular glycerol which otherwise could become toxic for the parasite due to its specific compartmentation of glycolysis. PMID- 9746371 TI - Shoulder dystocia. PMID- 9746372 TI - Medicolegal commentary: shoulder dystocia. PMID- 9746373 TI - Medicolegal issues in fertility regulation. PMID- 9746374 TI - The effect of pregnancy on survival in women infected with HIV: a systematic review of the literature and meta-analysis. AB - OBJECTIVE: To investigate the effect of pregnancy on disease progression and survival in women infected with HIV by a systematic review of the literature and meta-analysis. METHODS: Appropriate publications were identified using electronic and hand searching of relevant journals from 1983 to 1996. Studies were included in the review if they were cohort studies, either prospective or retrospective, or case-control studies which investigated disease progression of pregnant women infected with HIV and included a control group of non-pregnant women infected with HIV for comparison. Methodological quality was assessed for each study. Data were extracted for predetermined outcome measures. Sensitivity analyses were performed to explore the association between pregnancy and disease progression for the following study characteristics: clinical setting (developed or developing countries), methodological quality (high or poor) and whether studies had controlled for potential confounding. RESULTS: Seven studies, all prospective cohorts, were eligible to be included in the review. The summary odds ratio for the risk of an adverse maternal outcome related to HIV infection and pregnancy were as follows: death 1.8 (85% CI 0.99-3.3); HIV disease progression 1.41 (95% CI 0.85-2.33); progression to an AIDS-defining illness 1.63 (95% CI 1.00-2.67) and fall of CD4 cell count to below 200 x 10(6)/L 0.73 (95% CI 0.17-3.06). Sensitivity analyses showed that HIV progression in pregnancy was significantly more common in a developing country setting (odds ratio 3.71, 95% CI 1.82-7.75) than in developed countries (odds ratio 0.55, 95% 0.27-1.11) and also significantly more common in high quality studies when compared to low quality ones, odds ratios 3.71 (95% CI 1.82-7.57) and 0.55 (95% CI 0.27-1.11), respectively. However, there appears to be less progression of HIV disease and progression to AIDS when studies attempted to control for confounding by matching or restriction techniques, although this was not statistically significant in either case. CONCLUSIONS: The findings of this review have implications for women infected with HIV who are pregnant or are considering a pregnancy. There does appear to be an association between adverse maternal outcomes and pregnancy in women infected with HIV, although this association is not strong. The relation may be due to the result of bias including residual confounding. Further large scale observational studies with long term follow up are required before this issue can be fully resolved. PMID- 9746375 TI - The association between maternal HIV infection and perinatal outcome: a systematic review of the literature and meta-analysis. AB - OBJECTIVE: To investigate the association between maternal HIV infection and perinatal outcome by a systematic review of the literature and meta-analysis. METHODS: Appropriate publications were identified using electronic and hand searching of relevant journals from 1983 to 1996. Studies were included in the review if they were prospective cohorts with pregnant women identified as being HIV-infected with a control group of pregnant women who were not infected with HIV. Methodological quality was assessed for each study. Data were extracted for pre-determined outcome measures. Sensitivity analyses were performed to explore the association between HIV infection and an adverse perinatal outcome for the following study characteristics: clinical setting (developed or developing countries), methodological quality (high or poor) and whether studies controlled for potential confounding. RESULTS: Thirty-one studies were eligible to be included in the review. The summary odds ratio of the risk of pre-defined adverse perinatal outcomes related to maternal HIV infection were as follows: spontaneous abortion 4.05 (95% CI 2.75-5.96); stillbirth 3.91 (95% CI 2.65-5.77); fetal abnormality 1.08 (95% CI 0.7-1.66); perinatal mortality 1.79 (95% CI 1.14-2.81); neonatal mortality 1.10 (95% CI 0.63-1.93); infant mortality 3.69 (95% CI 3.03 4.49); intrauterine growth retardation 1.7 (95% CI 1.43-2.02); low birthweight 2.09 (95% CI 1.86-2.35) and pre-term delivery 1 83 (95% CI 1.63-2.06). Sensitivity analyses showed that the association between infant mortality and maternal HIV infection was stronger in studies conducted in developing countries when compared with developed countries [odds ratios (OR) 3.72 (95% CI 3.05-4.54) and 8.6 (95% CI 0.53-141.05), respectively]; studies of higher methodological quality compared with those of poorer quality [odds ratios 14.57 (95% CI 6.93 30.65) and 3.37 (95% CI 2.74-4.14), respectively] and studies which had used restriction or matching to control for potential confounding factors compared with those that had not [OR 11.60 (95% CI 5.71-23.58) and 3.35 (95% CI 2.73 4.12), respectively]. CONCLUSIONS: The findings of this review have implications for women infected with HIV who are planning a pregnancy or who find themselves pregnant. There appears to be an association, although not strong, between maternal HIV infection and an adverse perinatal outcome. This relationship may be due to bias including uncontrolled or residual confounding. There does, however, appear to be a real and large increase in the risk of infant death in developing countries associated with maternal HIV infection, especially so when there has been an attempt to control for confounding. PMID- 9746376 TI - Women's knowledge and attitudes, and the acceptability of voluntary antenatal HIV testing. AB - OBJECTIVE: To assess pregnant women's knowledge of, and attitudes towards, antenatal HIV testing, and its acceptability to them. SETTING: Antenatal clinic at Guy's Hospital, London, six community antenatal clinics and a midwifery group practice. POPULATION: Eight hundred and forty-three women attending the antenatal clinics. METHOD: The women received a leaflet explaining HIV testing, and completed a questionnaire before and after their booking appointment. This included an assessment of their knowledge of, and attitudes towards HIV testing, and its acceptability. RESULTS: Seven hundred and eighty-nine women (94%) completed questionnaires. Fifty-one percent (n = 405) were Caucasian, 25% (n = 195) African, 11% (n = 86) West Indian and 13% (n = 100) were from other ethnic groups. Fifty-eight percent received the HIV information leaflet, of whom 86% had read it. Knowledge relating to HIV was good, the median knowledge score being 6 out of a possible 8, but it was less in non-Caucasian women and those with lower educational qualifications. Knowledge was not related to uptake of testing. Thirty-five percent of women accepted the offer of an HIV test, rates being higher in hospital clinics (41%) than in the midwifery group practice (10%) and the community clinics (30%). Women more likely to accept the offer of an HIV test were non-Caucasian (P = 0.0443), those who had thought about the HIV test before this pregnancy (P = 0.0298) and those seeing one particular midwife (P = 0.0003). Most women (67%) thought that all pregnant women should be offered the HIV test and then make their own decision. Overall, 64% women did not change their original pre-discussion decision on testing for HIV. Thirty-six percent of women changed their decision from 'yes' to 'no' or 'don't know' after seeing the midwife. Women attending the community clinics (P = 0.003) and those who had been tested before (P = 0.0451) were more likely to change their decision. CONCLUSION: This study, in a multiethnic population, has shown that knowledge regarding HIV is good but does not increase the uptake of testing. Women prefer to be offered the HIV test and make their own choice regarding whether to accept it. PMID- 9746377 TI - The value of screening for Down's syndrome in a socioeconomically deprived area with a high ethnic population. AB - OBJECTIVE: To assess the utility of biochemical antenatal screening for Down's syndrome in a socioeconomically deprived area with a high proportion of Asian women from the Indian Subcontinent. DESIGN: Audit of Down's syndrome biochemical screening service over a four-year period. SETTING: Teaching hospital and community antenatal clinic in inner city Birmingham. POPULATION: Women booked between October 1992 and December 1996. METHODS: Blood for screening was collected between 14 and 21 weeks gestation, alpha-fetoprotein and intact human chorionic gonadotrophin were measured in serum and the risk of Down's syndrome was calculated. MAIN OUTCOME MEASURES: Uptakes of screening and amniocentesis, screen positive rate, odds of being affected given a positive result, miscarriages associated with amniocentesis offered following a high risk result, detection rate, number of Down's cases prevented and a cost analysis. Outcome measures were compared between Asians and Caucasians. RESULTS: Overall 11,974 women (71%) accepted serum screening. The screen positive rate was 8.3% in Asians and 5.0% in Caucasians. The uptake of amniocentesis in women following a high risk result was 54% overall (35% Asian, 67% Caucasian). Nineteen cases of Down's syndrome were identified, of which 13 occurred in women who opted for biochemical screening. The detection rate of the biochemical screening programme was 85% (11/13). Of these 11 cases, six (none of whom were Asian) elected to have an amniocentesis, of whom four thereafter had a termination. CONCLUSION: In this study the public health benefits of screening for Down's syndrome in a socioeconomically deprived area with a high Asian population, were small. PMID- 9746378 TI - Effect of routine screening for Down's syndrome on the significance of isolated fetal hydronephrosis. AB - OBJECTIVE: To determine the risk of Down's syndrome in fetuses with isolated hydronephrosis at 18-23 weeks in an unselected general population after routine screening for Down's syndrome, using first trimester nuchal translucency measurement and second trimester maternal serum biochemistry. POPULATION: All pregnant women undergoing a routine 18-23 week ultrasound scan, from a population who had been offered screening for Down's syndrome. SETTING: A district general hospital serving a low risk obstetric population. METHODS: Prospective study of all routine 18-23 weeks ultrasound scans. The prevalence of isolated hydronephrosis and Down's syndrome was determined and the relative risk for Down's syndrome was calculated for different ultrasound findings. RESULTS: 10,971 women were scanned at 18-23 weeks during the study period. Down's syndrome was diagnosed in 14 of 20 cases before this stage using first trimester nuchal translucency measurement and second trimester maternal serum biochemistry. Isolated fetal hydronephrosis was diagnosed in 423 pregnancies (3.9%); none of these pregnancies were affected by Down's syndrome. The relative risk for Down's syndrome was 0.18 (95% CI 0.06-0.53) for women with a normal scan (n = 9983). When multiple ultrasound markers were found (n = 565), the relative risk for Down's syndrome was 2.00 (95% CI 0.18-22.10) and 9.00 (95% CI 1.14-71.30) for all other aneuploidies. CONCLUSION: The finding of isolated fetal hydronephrosis does not significantly increase the age-related risk for Down's syndrome. The presence of multiple ultrasound markers is associated with an increased risk of aneuploidies other than Down's syndiome. These findings are explained by the reduced prevalence of Down's syndrome as a result of prior screening and diagnosis of this condition. PMID- 9746379 TI - Obstetric risk factors for periventricular leukomalacia among preterm infants. AB - OBJECTIVE: To evaluate the obstetric antecedents of cystic periventricular leukomalacia and transient echodense periventricular lesions among preterm infants. DESIGN: A cohort study of preterm singleton infants born between 25 and 33 weeks gestation. SETTING: Pavia, Italy. POPULATION: Three hundred and forty nine infants admitted to a Division of Neonatal Intensive Care who were screened for periventricular leukomalacia. METHOD: The obstetric factors in infants with either cystic periventricular leukomalacia or transient echodense periventricular lesions were compared to those in infants with negative cranial ultrasonographic findings. Stepwise multiple logistic regression analysis was used to evaluate the association between risk factors and outcomes adjusting for confounders. RESULTS: The prevalence of cystic periventricular leukomalacia and transient echodense lesions was 5.7% (20/349) and 14% (49/349), respectively. The main risk factors for cystic leukomalacia were first trimester haemorrhage (OR 4.49; 95% CI 1.63 12.39), maternal urinary tract infection on admission (OR 5.71; 95% CI 1.91 17.07), and neonatal acidosis (pH < 7.2) at birth (OR 5.97; 95% CI 1.93-18.52). Meconium-stained amniotic fluid (OR 3.95; 95% CI 1.42-10.98) and long term (> 72 hours) ritodrine tocolysis (OR 2.54; 95% CI 1.28-5.05) were associated with an increased risk of echodense lesions. The likelihood of overall leukomalacia (cystic plus echodense periventricular lesions) was increased among cases with meconium-stained amniotic fluid (OR 4.06; 95% CI 1.65-10.0), long-term ritodrine tocolysis (OR 2.56; 95% CI 1.38-4.72), maternal infection (OR 1.73; 95% CI 1.0 3.0), and acidosis at birth (OR 1.98; 95% CI 1.0-3.98). CONCLUSIONS: This study confirms that maternal infection, acidosis at birth, and meconium-stained amniotic fluid increase the risk of periventricular leukomalacia in preterm infants. Long-term ritodrine use seems to increase the risk for transient echodense lesions. PMID- 9746380 TI - The effect of fetal neck position on nuchal translucency measurement. AB - OBJECTIVE: To determine the influence of the position of the fetal neck on nuchal translucency measurement. DESIGN: A prospective cross-sectional study. POPULATION: One hundred and ninety-six. METHODS: Nuchal translucency was measured in the mid-sagittal plane, with the fetal neck in the flexed, neutral and extended positions. Measurements were made to the nearest 0.1 mm. Statistical analysis used the paired t-test for differences between the extended and neutral positions, [delta extended nuchal translucency] and the flexed and neutral positions [delta flexed nuchal translucency]. RESULTS: The mean extended nuchal translucency was 0.62 mm greater than the mean neutral nuchal translucency value [95% confidence interval 0.53 to 0.70, T = 14.33, P < or = 0.00001]. The mean flexed nuchal translucency was 0.40 mm less than the mean neutral nuchal translucency value [95% CI 0.34 to 0.47, T = 11.99; P = < 0.00001]. The repeatability coefficient was lower in the case of neutral nuchal translucency measurements [0.48] and was higher in the other groups [extended = 1.04, flexed = 0.70]. CONCLUSION: Fetal neck position can make a significant difference to nuchal translucency measurements. Repeatability of measurements are more accurate with the fetal neck in the neutral position. These findings have important implications for clinicians using nuchal translucency to screen the general obstetric population. PMID- 9746381 TI - Delivery outcome after the use of acid-suppressing drugs in early pregnancy with special reference to omeprazole. AB - OBJECTIVE: To study delivery outcome after maternal use of acid-suppressing drugs during early pregnancy. DESIGN: Cohort study of women identified by interview in early pregnancy. POPULATION: Sweden women giving birth from 1995 to early 1997. METHODS: Comparison of infants exposed to acid-suppressing drugs with all births in 1995-1996. MAIN OUTCOME MEASURES: Presence of congenital malformations. RESULTS: Proton pump blockers were used by 275 women, H2 receptor antagonists by 255 women, and both categories of drugs by 20 women. No effect of the use of omeprazole or H2-receptor antagonists on the rate of congenital malformations could be demonstrated. CONCLUSIONS: Though a teratogenic effect of these drugs cannot be completely ruled out, the individual risk after exposures during the first trimester seems to be negligible. PMID- 9746382 TI - The outcomes of pregnancy in women exposed to newly marketed drugs in general practice in England. AB - OBJECTIVE: To determine the proportion and nature of congenital anomalies in babies born to women exposed to newly marketed drugs during the first trimester. DESIGN: Non-interventional observational cohort studies. METHODS: The women were identified in confidence by the Prescription Pricing Authority. The doctor was sent a questionnaire to determine clinical events, including pregnancy, occurring after the drug was dispensed. A supplementary questionnaire determined the outcome of each reported pregnancy. SETTING: General medical practice in England. POPULATION: Women exposed to newly marketed drugs in whom pregnancy was recorded. MAIN OUTCOME MEASURES: Outcomes of pregnancies, the proportion and nature of congenital anomalies in the babies born. RESULTS: 2511 pregnancies were reported. In 831 of these pregnancies a newly marketed drug had been taken during the first trimester and in 74 during the second/third trimester. The outcome was ascertained for 780 (94%) of these 831 pregnancies: 547 (66%) births; 10 (1%) ectopic pregnancies; 94 (11%) spontaneous miscarriages; 5 (< 1%) missed abortions; 120 (14%) legal abortions; and 4 (< 1%) intrauterine deaths. 557 infants were born, of whom 14 (2.5%) had congenital anomalies. CONCLUSIONS: The proportion of live infants with a congenital abnormality born to mothers exposed to newly marketed drugs in the first trimester was similar to the percentage of congenital anomalies estimated by the Office for National Statistics. These data add valuable information to the safety database of these drugs. PMID- 9746383 TI - Risk of myocardial infarction, angina and stroke in users of oral contraceptives: an updated analysis of a cohort study. AB - OBJECTIVES: To investigate risk of myocardial infarction, angina and stroke in users of contraceptive pills compared with users of other methods of contraception. DESIGN: Prospective cohort study, with recruitment between 1968 and 1974 and annual follow up until the age of 45 years. After this age, only women who had never used oral contraception or those who had used it for eight or more years continued to be followed up annually until July 1994. SETTING: Seventeen family planning clinics in England and Scotland. POPULATION: 17,032 women aged between 25 and 39 years at entry to the study. MAIN OUTCOME MEASURES: Occurrence of angina, myocardial infarction or stroke that was associated with either hospital admission or outpatient referral to hospital or death. RESULTS: Increased risk of myocardial infarction in oral contraceptive users was observed only in women who were heavy smokers at entry to the study. In this subgroup the relative risk of a myocardial infarction was 4.2 (95% CI 1.4-16.6) in ever users of oral contraception compared with non-users, 4.9 (1.2-23.6) in current users, and 4.0 (1.3-16.2) in ex-users. In all current users the relative risk of angina was 0.5 (0.1-1.4), and the relative risk of ischaemic stroke was 2.9 (1.3-6.7). The increased risk of ischaemic stroke did not persist in ex-users. CONCLUSIONS: Use of oral contraception is associated with increased risk of ischaemic stroke and increased risk of myocardial infarction (only in heavy smokers), but no increased risk of angina. These increased risks need to be considered within the context of the very low absolute risks of cardiovascular disease in this population. 5880 women need to take oral contraception for one year to cause one extra stroke, and 1060 women who are heavy smokers need to take it for one year to cause one extra myocardial infarction. PMID- 9746384 TI - Risk of hysterectomy after 1000 consecutive endometrial laser ablations. AB - OBJECTIVES: To determine the hysterectomy rate after endometrial laser ablation, allowing for variable follow up times, and to evaluate the factors that might predict outcome. DESIGN: Observational cohort study. SETTING: Specialist minimal access gynaecology unit in a district general hospital. METHODS: Data were obtained from case notes, theatre records, and follow up postal questionnaires. The risk of hysterectomy following endometrial laser ablation was assessed using survival curve estimates. Proportional hazards regression analysis was used to identify the predictor(s) of this outcome. RESULTS: A single endometrial laser ablation was carried out on 746 patients (85.4%); 124 patients (14.2%) underwent one repeat procedure and three (0.4%) underwent two repeat ablative procedures. The cumulative rate of return of the postal questionnaires was 87.3% (762/873 patients). Survival curve analysis showed that the overall hysterectomy rate projected over a follow up period of 6.5 years was 21% (95% CI 16%-27%). The age of the patient at endometrial ablation, uterine cavity length, operative time, volume of fluid absorbed, the presence or absence of dysmenorrhoea, premenstrual syndrome and the method of endometrial preparation prior to surgery did not contribute significantly to the regression model. Having a repeat endometrial ablation procedure increased the risk of having a subsequent hysterectomy (RR = 2.93; 95% CI 1.59-5.40; P = 0.0015), whereas the presence of intrauterine pathology (eg, polyps, fibroids and uterine shape abnormalities) decreased the risk of this outcome (RR = 0.26; 95% CI 0.08-0.86; P = 0.0082) after adjustment for confounding due to patient's age and dysmenorrhoea prior to surgery. CONCLUSIONS: Endometrial laser ablation is a safe and effective treatment for menstrual dysfunction. Repeat ablative procedures significantly increased, and the presence of intrauterine pathology decreased, the risk of subsequent hysterectomy. PMID- 9746386 TI - Radical trachelectomy: a way to preserve fertility in the treatment of early cervical cancer. AB - In the Centenary year of Wertheim's hysterectomy for the treatment of invasive cervical cancer, it is appropriate to look at less radical methods of managing early stage disease. Radical trachelectomy with pelvic lymphadenectomy is a conservative but locally radical procedure, preserving the corpus uteri and therefore fertility potential. The first 10 cases in a pilot study are presented. One patient has required post-operative radiotherapy and another a completion radical hysterectomy. Three live births by caesarean section and three other pregnancies have resulted. Careful selection within strict criteria may allow this more conservative approach without compromising cure. These procedures should be carried out in referral centres with continuing follow up and review. PMID- 9746385 TI - A double-blind, randomised trial comparing the effects of tibolone and continuous combined hormone replacement therapy in postmenopausal women with menopausal symptoms. AB - OBJECTIVE: To compare the effects of two postmenopausal regimens on menopausal symptoms, bleeding episodes, side effects and acceptability. DESIGN: Double blind, randomised controlled trial. SETTING: Twenty-nine sites in Denmark, nine in Norway and six in Sweden. PARTICIPANTS: Four hundred and thirty-seven postmenopausal women with menopausal complaints. None of these women had had a hysterectomy. INTERVENTIONS: Daily treatment with tibolone 2.5 mg (n = 218) or 17beta-oestradiol 2 mg plus norethisterone acetate 1 mg (E2/NETA) (n = 219). MAIN OUTCOME MEASURES: Hot flushes, sweating episodes, vaginal dryness, assessment of sexual life and bleeding patterns; at baseline and after 4, 12, 24 and 48 weeks. RESULTS: Treatment with either preparation significantly reduced mean scores for hot flushes, sweating episodes and vaginal dryness. The overall discontinuation rate was 28% (tibolone 25%, E2/NETA 31%; P = 0.14), mostly during the first six months. There was a markedly lower cumulative incidence of bleeding or spotting episodes with tibolone compared with E2/NETA (P < 0.0001), mainly during the first six treatment cycles. CONCLUSIONS: Both tibolone and E2/NETA effectively alleviate menopausal symptoms. However, tibolone caused significantly fewer bleeding or spotting episodes, which were reflected by lower overall rates of bleeding, as well as lower drop-out rates due to bleeding. PMID- 9746387 TI - First trimester screening for Down's syndrome using maternal serum PAPP-A and free beta-hCG in combination with fetal nuchal translucency thickness. AB - The aim of this study was to evaluate the potential effectiveness of maternal serum pregnancy-associated plasma protein A (PAPP-A) and free beta-hCG in combination with nuchal translucency thickness in first trimester screening for Down's syndrome. Maternal serum levels of PAPP-A and free beta-hCG were assayed in stored sera from 32 Down's syndrome and 200 unaffected pregnancies. Fetal nuchal translucency was measured by ultrasound at the time of blood sampling. Screening of Down's syndrome using a combination of maternal age, PAPP-A, free beta-hCG and nuchal translucency would achieve a detection rate of 75.8% for a false positive rate of 5%. PMID- 9746388 TI - Eisenmenger's syndrome in pregnancy: maternal and fetal mortality in the 1990s. AB - To determine maternal and fetal mortality associated with Eisenmenger's syndrome in the UK, a postal questionnaire was sent to 225 NHS obstetric units with neonatal intensive care units, requesting information about maternal and fetal outcome in cases of Eisenmenger's Syndrome between 1991 and 1995. Fifteen cases were identified. The maternal mortality was 40% and fetal loss 8%. Only 15% of infants were born at term. Maternal mortality associated with Eisenmenger's syndrome remains as high as it has been for the past 50 years. Pooling of national data on rare medical conditions in pregnancy is required to aid management of individual cases. PMID- 9746389 TI - A mutation in the prothrombin gene contributing to venous thrombosis during pregnancy. PMID- 9746390 TI - Lymphangioma of the labia minora with deep lymphatic involvement. PMID- 9746391 TI - Hypersensitivity vasculitis (microscopic polyangiitis) in pregnancy with transmission to the neonate. PMID- 9746392 TI - A new questionnaire to assess the quality of life of urinary incontinent women. PMID- 9746393 TI - Pre-eclampsia: a state of sympathetic over-activity. PMID- 9746394 TI - Antenatal corticosteroid prescribing: setting standards of care. PMID- 9746395 TI - Blood splashes to the masks and goggles during caesarean section. PMID- 9746396 TI - Extra-amniotic saline infusion for induction of labour in antepartum fetal death: a cost effective method worthy of wider use. PMID- 9746397 TI - Maternal mortality--United States, 1982-1996. AB - Maternal and infant mortality are basic health indicators that reflect a nation's health status. In the United States, infant mortality has declined steadily; however, this is not true for maternal mortality. This report presents data from death certificates compiled by CDC's National Center for Health Statistics, which indicate that in the United States, the annual maternal mortality ratio remained approximately 7.5 maternal deaths per 100,000 live births during 1982-1996. PMID- 9746398 TI - Hepatitis A vaccination of men who have sex with men--Atlanta, Georgia, 1996 1997. AB - Outbreaks of hepatitis A among men who have sex with men (MSM) are a recurring problem in many large cities in the industrialized world. Because MSM are at high risk for acquiring hepatitis A, in 1995 the Advisory Committee on Immunization Practices (ACIP) recommended that MSM be vaccinated against hepatitis A. These recommendations have not been implemented widely, even in outbreak settings. This report summarizes the investigation of an ongoing outbreak of hepatitis A among MSM in Atlanta, Georgia, and a public health vaccination campaign in response to the outbreak. PMID- 9746399 TI - Effectiveness of a seventh grade school entry vaccination requirement--statewide and Orange County, Florida, 1997-1998. AB - Vaccine-preventable diseases continue to occur among adolescents (i.e., persons aged 11-21 years). In 1996, the Advisory Committee on Immunization Practices (ACIP), the American Academy of Pediatrics, the American Academy of Family Physicians, and the American Medical Association published joint recommendations emphasizing appropriate vaccination of adolescents aged 11-12 years who have not been vaccinated with hepatitis B vaccine, a second dose of measles, mumps, and rubella vaccine (MMR), varicella vaccine (if indicated), a booster dose of tetanus and diphtheria toxoids (Td), and other vaccines that may be indicated for certain adolescents. School entry requirements are an effective mechanism for ensuring high vaccination coverage among children. At the start of the 1997-98 school year, an amendment to the Florida Administrative Code (64D-3.011, F.A.C.) was instituted that requires all persons entering seventh grade to be vaccinated with three doses of hepatitis B vaccine, a second dose of MMR, and a Td booster, or to be on schedule for vaccination (i.e., having received at least one dose of hepatitis B vaccine, one dose of MMR, and a Td booster). To determine vaccination coverage among students entering seventh grade in Florida and in Orange County in 1997, CDC, in collaboration with the Florida Department of Health, analyzed state vaccination coverage data. This report summarizes the results of the analysis and indicates that a vaccination requirement for middle school entry can be effective in ensuring vaccination of adolescents. PMID- 9746400 TI - Recommendations of the Advisory Committee on Immunization Practices, the American Academy of Pediatrics, and the American Academy of Family Physicians: use of reminder and recall by vaccination providers to increase vaccination rates. AB - This statement by the Advisory Committee on Immunization Practices (ACIP), the American Academy of Pediatrics (AAP), and the American Academy of Family Physicians (AAFP) presents and recommends a programmatic strategy-the use of a reminder and/or recall (R/R) system by vaccination providers-to increase vaccination rates. In 1992, a national survey indicated that 8% of pediatricians and 5% of family physicians had implemented a manual vaccination R/R system and 6% and 5%, respectively, used a computer-based system for vaccination R/R messages. In 1993, the National Vaccine Advisory Committee issued the "Standards for Pediatric Immunization Practices," which recommend that all public and private health-care providers use a vaccination R/R system. These standards were endorsed by ACIP, AAP, and AAFP. By 1995 a survey indicated that R/R systems were used by 35% of pediatricians and 23% of family physicians (R. Zimmerman, University of Pittsburgh School of Medicine, personal communication, 1995). PMID- 9746401 TI - The POSNA pediatric musculoskeletal functional health questionnaire: report on reliability, validity, and sensitivity to change. Pediatric Outcomes Instrument Development Group. Pediatric Orthopaedic Society of North America. AB - The goal of orthopaedic interventions is to improve the functional health of patients, particularly physical function. The American Academy of Orthopaedic Surgeons and the Pediatric Orthopaedic Society of North America (POSNA) commissioned a work group to construct functional health outcomes scales for children and adolescents, focusing on musculoskeletal health. The work group developed scales assessing upper extremity function, transfers and mobility, physical function and sports, comfort (pain free), happiness and satisfaction, and expectations for treatment. Parent and adolescent self-report forms were developed and tested on 470 subjects aged 2-18 years. The POSNA scales demonstrated good reliability, construct validity, sensitivity to change over a 9 month period, and ability to outperform a standard instrument, the Child Health Questionnaire physical functioning scale. They were useful for a wide variety of ages and diagnoses. They appear to be ideally suited for orthopaedic surgeons to assess the functional health and efficacy of treatment of their patients at baseline and follow-up. PMID- 9746403 TI - Chronic musculoskeletal pain in childhood. AB - We studied 73 children with chronic or recurrent musculoskeletal pain of > or = 6 weeks' duration. Thirty-six children had no identifiable organic etiology for their pain, with a minimum follow-up of 2 years for ongoing symptoms. Thirty seven children had an organic etiology for their pain. Use of an Inappropriate Symptom Checklist was helpful in distinguishing between children with chronic pain who were found to have an organic disease and those without an identifiable organic disease. Seventy-seven percent of children with no inappropriate symptoms had an organic diagnosis ultimately made. Conversely, 79% of children with two or more inappropriate symptoms ultimately had no organic diagnosis to explain their pain. Behavioral self-report measures testing could not differentiate between children with chronic pain with or without organic disease. Intervention by a psychologist skilled in pain management was helpful. PMID- 9746402 TI - Orthopaedic manifestations of chronic graft-versus-host disease. AB - Chronic graft-versus-host disease (GVHD) is a well-recognized complication of allogeneic bone marrow transplantation (BMT). Musculoskeletal manifestations include joint contractures, polymyositis, polyserositis, and fasciitis. We present 14 patients with orthopaedic complications of chronic GVHD. Long-term conservative management of joint contractures with physical therapy and orthotics was generally successful in restoring patients' premorbid functional status. Surgical release of joint contractures yielded poor results and rendered the affected joints unresponsive to further conservative treatment. Surgical intervention in the treatment of joint contractures resulting from chronic GVHD does not appear qualitatively to improve functional status in patients affected with this disease process. PMID- 9746404 TI - Micromechanical properties of epiphyseal trabecular bone and primary spongiosa around the physis: an in situ nanoindentation study. AB - The elastic modulus and hardness of the mineralized bone around the growth plate was measured to determine its regional micromechanical properties. Multiple nanoindentation tests, >10 sessions, with depths ranging from 100 to 1,000 nm at loading rates of 12.5 and 750 microN/s, were performed on the trabecular bone in the epiphysis, trabecular bone at the junction of the physis and epiphysis, primary spongiosa in the metaphysis, and surrounding cortical bone of the distal femur of 300-gm Sprague-Dawley rats. The indentation load-displacement data obtained in these tests were analyzed to determine the elastic modulus and hardness of the tissues. The nanoindentation results highlighted the regional variations in the material properties of the mineralized tissues around the growth plate. The primary spongiosa had a lower elastic modulus and hardness than both epiphyseal trabecular and cortical bone (p < 0.01). A relatively well defined thick trabecular band at the physeal-epiphyseal junction had modulus and hardness values comparable to those of cortical bone (p > 0.05). These findings support the hypothesis that the primary spongiosa has micromechanical properties that are significantly lower than the epiphyseal trabecular bone. On this basis, it is speculated that the fracture patterns commonly seen in patients with physeal injuries are influenced by the micromechanical properties of these tissues, as well as by the nature and direction of the applied force. PMID- 9746405 TI - Callus response to micromovement during elongation in the rabbit. AB - The purpose of this investigation was to determine whether induced micromovement during the elongation period could improve the consolidation of diaphyseal elongation obtained by callus distraction. Two series of paired rabbit hindlimbs were studied. The surgical procedure and the waiting period were identical. During elongation, one hindlimb was stimulated, and the other was the control. The consolidation period was 2 days. Reproducible tibial osteotomy and lengthening of the two tibiae was confirmed radiographically. The mineralized callus was quantified by dual-beam x-ray absorptiometry. The callus diameters were measured. Bones were axially compressed to failure. Callus volume, mineral quantity, mineral density, and resistance to failure were not different on the stimulated side compared with the unstimulated side, so micromovement applied during elongation had no effect on bone consolidation. For all tibiae, resistance to failure of the callus was significantly correlated to callus volume, to callus mineral content, and to callus mineral density. PMID- 9746406 TI - A method for normalization of oxygen cost and consumption in normal children while walking. AB - Measurement of oxygen use is helpful in determining energy consumption in children with walking abnormalities; however, no statistically valid measurements of nondisabled children have been established using a telemetric system. Data from 94 nondisabled children, ages 5-15 years, were collected using the Cosmed K2 oxygen analysis system. Oxygen cost, measured in milliliters O2/kg/m walked, and oxygen consumption, measured in milliliters O2/kg/min, were correlated to inverse body surface area (IBSA) measured in meters(-2). Linear relationships between oxygen cost and IBSA and between oxygen consumption and IBSA were best described by the following equations: oxygen cost = 0.256 (IBSA) + 0.052 (r = 0.806) and oxygen consumption = 17.635 (IBSA) + 4.956 (r = 0.758). From these data, equations were derived to calculate predicted oxygen cost and predicted oxygen consumption for each child. Indices were developed to express the difference between a measurement and the predicted mean in reference to the normal variation. These equations and indices can help quantify the variation of energy use of children with walking abnormalities when compared with their nondisabled peers. Additionally, the indices enable multiple tests from one subject to be compared, regardless of a change in age, height, and weight between measurements. PMID- 9746407 TI - Hematuria associated with low-volume cell saver in pediatric orthopaedics. AB - A low-volume autotransfusion device, Haemocell System 350, was used for four consecutive pediatric orthopaedic patients. Although the initial patient evidenced no hematuria, transient hematuria was noted in three consecutive patients. Follow-up blood urea nitrogen, electrolytes, and creatinine levels were all within normal limits. After discontinuation of the device, no further hematuria has occurred in subsequent patients. Although intraoperative low-volume cell savers may have a role in pediatric orthopaedic surgery associated with low total blood volume loss, we observed three cases of postoperative hematuria by using this device in cases with large total blood volume loss [>28% estimated blood volume (EBV)]. Although all of these cases were transitory, we recommend caution in the use of low-volume intraoperative blood-salvage devices in pediatrics at this time. PMID- 9746408 TI - Surgical correction of muscular torticollis in older children with Peter G. Jones technique. AB - In the Department of Pediatric Surgery, Uludag University Medical Faculty in Bursa, during the last 11 years, the Peter G. Jones technique for the surgical correction of muscular torticollis in older children has been introduced. Twenty children between 4 and 13 years of age were treated for muscular torticollis. They were followed up from 3 months to 10 years after surgery. All patients had a middle-third open transection of the sternocleidomastoid muscle. Preoperative and postoperative assessment by a rigid scoring system showed that all patients improved in terms of function as well as cosmesis. Children younger than 10 years showed the most improvement, with 90% excellent and good results. Late middle third open transection of the sternomastoid in muscular torticollis may give acceptable results. PMID- 9746409 TI - Estimation of the lumbar curve magnitude with correction of the right thoracic curve in idiopathic scoliosis. AB - A simple formula was proposed to estimate the magnitude of the postoperative uninstrumented lumbar curve with correction of the right thoracic curve in idiopathic scoliosis. This formula is as follows: PLC < or = LC - 0.5(TC - BTC) (PLC, predicted postoperative standing lumbar Cobb angle; LC, preoperative standing lumbar Cobb angle; TC, preoperative standing thoracic Cobb angle; BTC, preoperative supine lateral bending thoracic Cobb angle). Sixty-five patients' preoperative and postoperative radiographic measurements were taken, and of these 45 had measurements taken after > or = 12 months of follow-up. Multiple regression (R) value for the proposed formula postoperatively was 0.8048 and at > or = 1 year follow-up was 0.6869. PMID- 9746410 TI - Familial synspondylism: progressive scoliosis and multiple hernias in a kinship. AB - A new genetic syndrome is reported of congenital lordoscoliosis due to lumbar segmentation defects and incomplete formation of lumbar vertebrae. The defect arose as a spontaneous mutation and was transmitted in an autosomal dominant fashion. The kindred included a mother and her three offspring. These affected individuals had several dysmorphic features including cavus feet and micrognathia. In addition the syndrome was associated with multiple hernias including inguinal, ventral, and diaphragmatic. These associated problems led to the early death of the first child at age 7 months. The lumbar scoliosis was already evident by that time. The progressive nature of the scoliosis was documented, especially in one child who was lost to follow-up and who was initially seen with a severe spinal deformity. Surgical management was required in members of the kindred, but because of differences in age and severity at the time of surgery, the techniques varied. PMID- 9746411 TI - Posterior dislocation of the humeral head in association with obstetric paralysis. AB - Twelve children with obstetric paralysis were diagnosed as having a posterior dislocation of the humeral head. The diagnosis was suspected on clinical grounds and confirmed by computed tomography (CT) scans in all cases. All 12 patients were treated with open reduction via an anterior approach. The age range at the time of surgery was from 7 months to 7 years (average, 2 years and 3 months). All patients were immobilized in a shoulder spica for 6 weeks and a further 6 weeks in an orthosis. All patients were examined by CT scans in the postoperative period, which confirmed a satisfactory reduction in all cases. With a minimal follow-up of 12 months, there have been no redislocations. This article demonstrates that dislocation of the shoulder in association with obstetric paralysis is not rare, as previously described, and shows that once diagnosed, the dislocation can be satisfactorily treated by a single anterior open reduction of the shoulder. PMID- 9746412 TI - Autogenic bone marrow injections as a treatment for simple bone cyst. AB - Simple bone cyst (SBC) is a benign fluid-filled cavity found primarily at the proximal ends of long bones in children. Treatments proposed for SBC range from observation to intralesional curettage and bone grafting, which are all associated with uncertainty and complications. Because of these factors, a relatively noninvasive protocol with osteoinductive autogenic bone marrow was instituted. Twelve patients were identified with SBCs. Bone marrow was aspirated from the patient's iliac crests and injected into the cyst cavity. Follow-up ranged from 9 to 57 months. Eight (67%) patients demonstrated substantial healing, two (17%) showed partial healing, and two (17%) did not respond to bone marrow therapy. The advantages suggested by bone marrow injection over the currently practiced methods include a higher success rate with a single injection and earlier healing. PMID- 9746413 TI - Treatment of unicameral bone cyst with demineralized bone matrix. AB - Eleven patients with active unicameral bone cysts were treated primarily with placement of demineralized bone matrix in the cyst by using a two-needle technique and a custom large-bore needle. Cyst healing was rated according to the Neer classification, and the average time of healing was 4.5 months. The demineralized bone matrix demonstrated an ability to obliterate the cyst in nine of 11 patients by using a single injection within 4-5 months, and at 2 years' follow-up, no cysts were deemed active or recurrent. PMID- 9746414 TI - The limping child: a manifestation of acute leukemia. AB - Nine patients who presented to our institution with the chief complaint of a limp and no history of trauma were subsequently diagnosed with leukemia. A review of these patients identified clinical and laboratory findings that helped to establish the diagnosis. The presence of an antalgic gait with complaints of pain of variable intensity and duration, an irritable hip or knee, a mild to moderate elevation in body temperature, lymphadenopathy, hepatosplenomegaly, an increased erythrocyte sedimentation rate, thrombocytopenia, anemia, decreased neutrophils, increased lymphocytes, or blast cells on the peripheral blood smear should cause the physician to suspect leukemia in a limping child. Bone marrow biopsy confirms the diagnosis. PMID- 9746415 TI - Slipped capital femoral epiphysis associated with radiation therapy. AB - We reviewed 32 children with 41 radiation-therapy associated slipped capital femoral epiphyses (RTASCFE). Ten were from the authors' institutions and 22 from the literature. Gender distribution was equal. The age at diagnosis of the malignancy was 4.3 +/- 3.1 years; the amount of radiation was 4,240 +/- 1,445 rads. Children with RTASCFE presented younger (10.4 +/- 3.2 years) than a routine SCFE. The average symptom duration was 5 +/- 6 months. Children with RTASCFE are usually thin (median weight, 10th percentile) in contrast to children with typical SCFE, who are usually obese (<95th percentile). The majority (82%) of the slips were mild, compared to routine SCFEs (approximately 50%); 28% were bilateral. There was a positive linear relationship between the age at presentation of the SCFE and the age at diagnosis of the malignancy; there was a negative linear relationship between the age at presentation of the SCFE and the amount of radiation therapy. PMID- 9746416 TI - Relationship between lateral subluxation and widening of medial joint space in Legg-Calve-Perthes disease. AB - Fifty-five patients (61 affected hips and 49 unaffected hips) with Perthes disease were reviewed to evaluate the relationship between widening of medial joint space and lateral subluxation of the femoral head in radiographs. The components of the medial joint space were evaluated by using T1, T2, proton, and Gd-enhanced T1WI magnetic resonance images (MRI). The widened medial joint space in radiographs was filled with overgrown cartilage at the initial stage (27 hips) in MRI, with both overgrown cartilage and widened true medial joint space at the fragmentation stage (23 hips) and widened true medial joint space at the healing stage (11 hips). Between affected hips and unaffected hips, the mean difference of medial joint space in radiographs between hips at the initial stage and at the fragmentation stage was 2 and 4.5 mm, respectively; the mean difference in percentage of lack of coverage of the femoral head between hips at the initial stage and at the fragmentation stage was 3 and 15%, respectively. During the healing stage, widening of the medial joint space decreased or normalized because of ossification of overgrown cartilage despite the existence of lateral subluxation because of coxa magna. We concluded that widening of the medial joint space may be used as an index of lateral subluxation at only the fragmentation stage in Perthes disease. PMID- 9746417 TI - Closed treatment of hip dislocation in Down syndrome. AB - Two young children (three hips) with Down syndrome and dislocation of the hip were successfully treated by nonoperative methods by using the principle of prolonged immobilization or bracing. A 5-year, 6-month-old patient with bilateral habitual dislocation used an ambulatory abduction orthosis full-time for 6 months and then part-time for 4 months. Complete dislocation of the right hip in a 4 year, 6-month-old patient was managed by closed reduction, spica cast immobilization for 4 months, and then an ambulatory abduction orthosis for 8 months. Both patients developed stable, well-contained hips. Nonoperative management of hip dislocation in Down syndrome can be successful and avoids the complications associated with operations previously recommended for these patients. PMID- 9746418 TI - Results of hip arthrodesis in adolescents by using the cobra-head plate for internal fixation. AB - A retrospective study was conducted to examine the efficacy and potential morbidity of hip fusion using the Cobra-head plate in adolescents. A heterogeneous group of 11 adolescents, with recalcitrant hip pain and an average age of 14.6 years, underwent unilateral hip arthrodesis with the Cobra-head plate for internal fixation. Seven (64%) patients achieved an uneventful clinical and radiographic arthrodesis after a mean postoperative interval of 7.4 months. Four index operations (36%) were complicated by pseudarthrosis. All patients who developed a postoperative pseudarthrosis ranked at or above the 90th percentile for their age-determined weight. The relationship between the percentile-weight for-age and the incidence of pseudarthrosis was statistically significant (p < or = 0.001). Hip-arthrodesis procedures with the Cobra-head plate in adolescents at or above the 90th percentile weight-for-age are associated with an unacceptably high rate of pseudarthrosis. In this subset of patients, alternate or supplementary stabilization methods should be considered. PMID- 9746420 TI - Acetabular volume. AB - Reconstructive acetabular osteotomies can affect the acetabular volume. Volume mismatch between the femoral head and the acetabulum should be an important consideration but is rarely evaluated before hip reconstruction. Accurate measurement of the volume of the acetabulum is difficult because of the unusual shape and spatial orientation of the acetabulum. In this study, we used three techniques (physical, two-dimensional computed tomography, and three-dimensional computed tomography reconstruction) to determine the volume of 18 pig, four sheep, and 15 model acetabulae. A comparison of pre- and post-Pemberton osteotomy volumes of three dysplastic acetabulae models and two patients with developmental dysplasia of the hip also was performed. The results indicate that accurate, reproducible volume determinations can be made by using all three techniques, and that certain data-selection modes can reduce the patient's exposure to radiation. In addition, we observed an increase in the volume of the acetabulum after the Pemberton osteotomy. PMID- 9746419 TI - Hip arthrodesis using the AO modular external fixator. AB - Arthrodesis remains the treatment of choice for children and young adults with deteriorated, painful hip joints. In 1992, Fernandez Dell'Oca et al. reported on the use of the AO external fixator in hip arthrodesis. Three patients who underwent this procedure are reported. Hip arthrodesis with AO modular external fixator has all the benefits of the Thompson arthrodesis, with the added advantages of producing a solid fusion in the correct position and allowing the patient to remain ambulatory while using the external fixator. PMID- 9746421 TI - Separation of the proximal femoral epiphysis after derotation varus osteotomy of the femur. AB - Proximal femoral varus and derotation osteotomy is a common procedure performed in the management of developmental dysplasia of the hip. This procedure imposes high shear stress on the femoral epiphysis, depending on the degree of varus obtained. We report two cases of proximal femoral epiphyseal slip after varus derotation osteotomy and discuss the management and outcome. Such epiphyseal slip may or may not be symptomatic, and a careful radiologic examination should be carried out in suspected cases. Management should be individualised. Surgical correction of varus may be required. PMID- 9746422 TI - Standing is a causative factor in osteonecrosis of the femoral head in growing rats. AB - We studied femoral head lesions resulting from applying excessive mechanical stress to the hip joints of ordinary growing Wistar Kyoto rats by forcing them to stand. Twenty rats were fed in high and low cages 5-15 weeks after birth. High rat cages were prepared with the feeding apparatus placed up high so that the rats had to stand on their hindlimbs to feed. In contrast, the rats in low cages could not stand up. Histological examination of the rats at 15 weeks showed that osteonecrosis of the femoral head and ossification disturbance occurred frequently in the high cage group. The incidence of osteonecrosis was 40% (eight of 20 femoral heads) and that of ossification disturbance was 10% (two of 20 femoral heads). In the femoral heads with these lesions, localized cartilaginous abnormalities were found frequently in the lateral portion of the femoral head, where the feeding vessels of the femoral head penetrate the epiphyseal nucleus. These findings were rarely seen in the low cage group. These results indicate that standing was a causative factor in osteonecrosis of the femoral head in growing rats. PMID- 9746423 TI - Orthotic treatment of infantile tibia vara. AB - Difficulty differentiating physiologic genu varum from early Blount's disease persists. Drennan's metaphyseal-diaphyseal (MD) angle remains the most consistently valuable radiographic parameter despite measurement error. Clinical risk factors also should be considered. All patients receiving orthoses for genu varum since 1985 were reviewed. The focus of the study was those patients with an MD angle of >16 degrees or between 9 and 16 degrees with a clinical risk factor for progression. Risk factors considered were ligamentous instability, obesity, asymmetry, and being female, black, or Hispanic. Thirty-eight patients with 60 tibiae were included. The success rate was 90%. Risk factors for failure (six cases) were instability, obesity, and delayed bracing. In cases with MD angles >16 degrees, the success rate was 86%. The results of orthotic treatment, restricted to patients meeting the stated parameters, represent improvement on the reported natural history. PMID- 9746424 TI - Correlation of MRI and arthroscopic diagnosis of knee pathology in children and adolescents. AB - The accuracy of magnetic resonance imaging (MRI) in diagnosing knee pathology in the pediatric and adolescent population is not well established. The purpose of this study was to correlate the findings of MRI and knee arthroscopy in children and adolescents. One hundred and eight consecutive knee arthroscopies performed in patients ages 4-17 years between 1992 and 1996 were retrospectively reviewed. Fifty-three of these patients underwent preoperative MRI. Age-related comparisons were then made between MRIs and observed intraoperative meniscal and anterior cruciate ligament pathology. The pediatric group (ages 4-14 years) was demonstrated to have an appreciable decrease in sensitivity, specificity, positive predictive value, and accuracy for essentially all categories of pathologic changes. Conversely, negative predictive values for the pediatric group exceeded those of the adolescent group (ages 15-17 years) in each category. The ability of MRI to predict intraarticular knee pathology among adolescents is comparable to that in adults, whereas it is much less accurate in the pediatric population. PMID- 9746425 TI - Evaluation of patella position based on radiologic and ultrasonographic examination: comparison of the diagnostic value. AB - This study compared radiologic and ultrasonographic methods of evaluation of patella position. The radiologic examination was based on the evaluation of Insall and Salvati's index (I-S index), whereas the ultrasonographic examination was based on the determination of analogous coefficient called the patellar tendon-patellar coefficient (T-P coefficient). The total number of examined knee joints was 55 in 30 patients (13 children, aged 7-16 years and 17 adults aged 17 39 years) with knee pain. Considerable differences of the evaluated parameters were observed in the group of examined children: I-S index, 1.50; T-P coefficient, 1.20; and small differences in the group of adults: 1.17 and 1.32, respectively. Those differences resulted from difficulties with interpretation of the apparent radiologic picture of the knee joint with the patella incompletely ossified. The ultrasonographic picture in both children and adults is a real picture, and the possibilities of its interpretation are independent of the degree of patellar ossification. PMID- 9746427 TI - Ulnar nerve injury after K-wire fixation of supracondylar humerus fractures in children. AB - Six cases of ulnar nerve injury resulted from crossed K-wire fixation of displaced supracondylar humeral fractures in children. The age ranged between 4 and 10 years. Pain on extension of the little and ring fingers and early clawing were important post operative signs of ulnar nerve involvement. Early exploration of all six cases revealed medial pin placement in the cubital tunnel in five cases. In two of these, the nerve was directly penetrated, and in three, it was constricted by the cubital tunnel retinaculum. In the case 6, the nerve was hypermobile and found to be fixed anterior to its groove over the medial epicondyle. The nerve was decompressed in all cases, and the wire was repositioned. Follow-up ranged from 4 to 14 months. Full nerve recovery occurred in three cases, partial in two, and no recovery in one. Early exploration rather than simple pin removal is safer and diagnostic of the mechanism of injury. PMID- 9746426 TI - Ulnar nerve injury after closed forearm fractures in children. AB - We treated two children with the unusual complication of ulnar nerve palsy after closed both-bone forearm fractures. Both patients developed an ulnar claw-hand deformity within 7 weeks of injury that resolved spontaneously by 20 weeks postinjury with nonoperative treatment. No patient showed any signs or symptoms of an ischemic compartment syndrome. Both nerve injuries were identified immediately at the time of fracture by a careful neurologic examination. This avoids confusion with a postreduction nerve entrapment injury or ischemic injury after a localized compartment syndrome, which may have considerably different treatments and outcomes. We recommend that a careful neurologic examination be recorded before any manipulative reduction of forearm fractures in children. If an ulnar nerve palsy is detected, it is probably a result of nerve contusion and should resolve without the need for surgical exploration. PMID- 9746428 TI - Traumatic hip dislocation in childhood. AB - Eighteen cases of traumatic dislocations of the hip in children under 15 years of age presenting between 1985 and 1995 were reviewed. Fifteen of the dislocations were posterior. There were two groups: a younger group dislocating with minimal trauma and an older group whose injuries involved significantly more trauma. All patients were treated by closed reduction, but two required open reduction because of intraarticular fragments preventing a full reduction. On long-term follow-up of the 16 available patients (average length of follow-up, 5 years 10 months; range, 17-132), there were no cases of avascular necrosis or early degenerative change. PMID- 9746429 TI - Ilizarov applications in the pediatric foot. PMID- 9746430 TI - Youth agricultural work-related injuries treated in emergency departments--United States, October 1995-September 1997. AB - National estimates and descriptions of agricultural injuries occurring to youths are limited. In 1996, the National Committee for Childhood Agricultural Injury Prevention recommended establishing and maintaining a comprehensive national surveillance system of fatal and nonfatal childhood agricultural injuries. In response to these recommendations, CDC's National Institute for Occupational Safety and Health (NIOSH) began analyzing existing surveillance data while exploring new data collection strategies. The goals of these efforts are to add to knowledge about the incidence and circumstances of childhood agricultural injuries and to improve collection and analysis of data regarding childhood agricultural injuries. This report presents an analysis of data from the National Electronic Injury Surveillance System (NEISS) during October 1995-September 1997 for youths aged <20 years, which indicates that youths in this age group are at increased risk for agricultural work-related injuries. PMID- 9746431 TI - Haemophilus influenzae invasive disease among children aged <5 years--California, 1990-1996. AB - Haemophilus influenzae (Hi) causes a variety of severe clinical illnesses including meningitis, pneumonia, epiglottitis, and septic arthritis. In the prevaccine era (i.e., before 1988), Haemophilus influenzae type b (Hib) caused approximately 95% of the Hi invasive disease among children aged <5 years. In 1988, Hib conjugate vaccines were introduced for use among children aged 18 months-5 years; they were subsequently recommended for routine use in infants by the Advisory Committee on Immunization Practices (ACIP) in 1990. During 1989 1995, Hib invasive disease among children aged <5 years declined 95% nationally. To document the decline of Hib invasive disease and to examine the epidemiology of reported nontype b Hi invasive disease among children aged <5 years, CDC, in collaboration with the California Department of Health Services, analyzed reported cases in California from 1990 to 1996. This report summarizes the results of the analysis and documents the decline of Hib without an increase of nontype b Hi invasive disease among children aged <5 years. PMID- 9746432 TI - Expression of pI(Cln) mRNA in cultured bovine lens epithelial cells: response to changes in cell volume. AB - PURPOSE: The authors recently established a link between swelling-activated myo inositol efflux and chloride movement via anion channels in cultured bovine lens epithelial cells (BLECs). To further define this pathway, the relationship between cell volume, myo-inositol movement and mRNA expression of pI(Cln), a proposed chloride channel regulatory protein was investigated. METHODS: To demonstrate the effect of cell volume changes on pIcln transcription, BLECs were exposed to either hypertonic or hypotonic medium conditions. For rapid cellular shrinkage, BLECs were maintained at confluence in physiologic medium (257+/-2 mosm) then transferred to sodium hypertonic medium (473+/-6 mosm) or raffinose hypertonic medium (452+/-2 mosm). For rapid cellular swelling, cells were switched from sodium hypertonic medium to physiologic medium+/-tamoxifen. The expression of pI(Cln) mRNA was determined by Northern blot analysis. RESULTS: Upon cell volume reduction (increasing intracellular osmolality), BLECs upregulate the expression of pI(Cln) mRNA. Contrastly, when cell volume rapidly increases (decreasing intracellular osmolality), BLECs moderately downregulate pIcln mRNA, with expression levels reaching near physiologic control by 24 h. Blockage of swelling-activated chloride movement and osmolyte efflux with either tamoxifen or niflumic acid enhances the downregulation of pIcln mRNA expression. CONCLUSIONS: In cultured BLECs, pI(Cln) transcriptional regulation appears to be responsive to cell volume fluctuations. These data suggest a converse relationship exists between pIcln mRNA expression and changes in cell volume. PMID- 9746433 TI - Cloning and expression of human corneal calgranulin C (CO-Ag). AB - PURPOSE: A host-parasite interaction is thought to be involved in the pathogenesis of Mooren's ulcer. We have identified a cornea-associated antigen (CO-Ag), which may be a target for the autoimmune process resulting in Mooren's ulcer. This study presents the cloning, expression, and identification of a cDNA encoding human CO-Ag. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) was performed to amplify a cDNA encoding CO-Ag in the human cornea. The cDNA fragment was cloned into a prokaryotic expression vector and the resulting plasmid was transformed into DH5 E. coli cells. Autoantibody reactivity to the CO Ag fusion protein in patient sera was tested by Western blots. RESULTS: A cDNA encoding human CO-Ag was amplified by RT-PCR. The entire mRNA coding region was 273 nucleotides in length, predicting a 91-amino acid protein with a molecular weight of 10,683 daltons. The cDNA sequence was identical to human neutrophil calgranulin C (CaGC). Human CO-Ag was expressed in E. coli carrying a plasmid in which the CO-Ag cDNA was under control of the E. coli trc promoter. The CO-Ag fusion protein, which comprised as much as 15% of the total bacterial protein, was purified to 90% homogeneity by affinity chromatography on an immobilized metal column. The recombinant CO-Ag protein produced was recognized by autoantibodies in the sera of 6 of 15 patients with Mooren's ulcer and none of 14 normal control sera by Western blots. CONCLUSION: CO-Ag is identical to calgranulin C, a neutrophil protein found on the surface of filarial nematodes. A host-parasite interaction may cause autoimmunity to CO-Ag (CaGC) in the cornea resulting in a Mooren's ulcer. PMID- 9746434 TI - Modulation of VEGF production by pH and glucose in retinal Muller cells. AB - PURPOSE: To investigate the influence of pH and glucose concentration, both of which represent significant biochemical variables in tissue ischemia, on the production of VEGF protein by retinal Muller cells and C6 glioma cells, under normoxic and hypoxic conditions. METHODS: Rat retinal Muller cells and C6 glioma cells grown in tissue culture monolayers were studied. The effect of pH (range 7.0-8.0) and glucose concentration (0.6-25 mmol/L) on VEGF protein production, under both normoxic and hypoxic conditions, were evaluated by ELISA analysis of the conditioned media. Establishment of significant cell hypoxia was verified by measurement of lactate release into the conditioned media. RESULTS: Hypoxia caused a 7.9-fold increase in VEGF production in C6 cells at 24 h, and a 3.4-fold increase in Muller cells after 48 h. Under hypoxic conditions, VEGF protein production was increased further by increasing pH and increasing glucose, and decreased by low pH and low glucose. Varying the glucose concentration or pH of the medium did not result in significant induction of VEGF protein production by either cell type under normoxic conditions. CONCLUSIONS: Both glucose and pH significantly affected VEGF production induced by low oxygen. However, neither exerted a measurable stimulatory effect on VEGF production in normoxic conditions. Coexisting hypoxia and acidosis or hypoglycemia, as might occur in severe tissue ischemia, may render glial cells incapable of effectively upregulating VEGF synthesis, while alkalosis or hyperglycemia may augment hypoxia induced VEGF production. PMID- 9746436 TI - The occurrence of retinol and carotenoids in human subretinal fluid. AB - PURPOSE: To investigate if retinol and carotenoids are present in the subretinal space following rhegmatogenous retinal detachment. METHODS: Blood and subretinal fluid were collected from patients at the time of surgical repair of retinal detachment. After removal of cellular contents in a specimen by centrifugation, the supernatant fraction was analyzed by liquid chromatography using a silica column eluted by 16% dioxane in hexane. Retinol and carotenoids were identified in the chromatograms based on their retention time and absorption spectrum. RESULTS: The retinol concentrations (mean+/-SD) in the serum and subretinal fluid were 305+/-144 and 166+/-96 ng/ml respectively. The 450 nm chromatogram had 7 peaks with the characteristic absorption spectrum of carotenoids. Peak 1 and 7 coincided with the retention time of beta-carotene (1.8 min) and lutein (10.8 min) respectively. The concentrations of beta-carotene and lutein in serum were 161+/-63 and 142+/-98 ng/ml respectively. There was very little beta-carotene in subretinal fluid (4.7+/-2.4 ng/ml). Lutein was the major carotenoid peak in subretinal fluid (41.4+/-14.1 ng/ml). The minor carotenoid peaks of serum were not observed in subretinal fluid. CONCLUSION: There is a substantial amount of retinol and lutein in subretinal fluid. The high proportion of lutein and very low amount of beta-carotene in the subretinal fluid support the occurrence of a highly selection transport mechanism of lutein from the blood to the retina. PMID- 9746435 TI - A mutation in the connexin 50 (Cx50) gene is a candidate for the No2 mouse cataract. AB - PURPOSE: The No2 cataractous mouse mutant displays a bilateral, congenital, hereditary nuclear opacity of the ocular lens. The aim of this work was to identify and subsequently screen an optimal candidate gene for a mutation correlated and consistent with the observed phenotype. METHODS: The No2 cataract was mapped in relation to genes and microsatellite markers by crossing to the wild mouse strain Mus spretus and then backcrossing to the inbred strain C3H/ HeH. The Cx50 (MP70) protein coding region and flanking sequences were amplified from normal parental as well as heterozygous and homozygous mutant genomic DNAs. These PCR products were then sequenced directly. Sequence data was corroborated by restriction analysis of PCR products. RESULTS: Mapping of the No2 cataract placed it in the vicinity of Gja8, the gene encoding connexin 50 (MP70), a major component of lens fiber gap junctions. Amplification and subsequent sequencing of the Cx50 protein coding regions revealed a single A-->C transversion within codon 47. This sequence change resulted in the creation of an HhaI restriction endonuclease restriction site, allowing for corroboration of the sequence data via restriction analysis using this enzyme. The sequence alteration is also predicted to result in the nonconservative substitution of alanine (Ala) for the normally encoded aspartic acid (Asp) at this position within the polypeptide. CONCLUSIONS: The identified mutation in Gja8 is both correlated and consistent with the cataract observed in the No2 mouse mutant, making it an ideal candidate for the cataract. This study provides the first evidence that a mutation in a lens connexin can result in congenital hereditary cataract, highlighting the importance of lens connexins in maintaining lens transparency. PMID- 9746437 TI - Chloride and sodium transport across bovine ciliary body/epithelium (CBE). AB - PURPOSE: To study the chloride and sodium ion transports across the bovine ciliary body/epithelium (CBE) by a modified Ussing-Zerahn type chamber. METHODS: Isolated bovine CBE preparations were mounted in a modified Ussing-type chamber and the transepithelial electrical parameters were monitored. The inward (stroma to aqueous) and outward (aqueous to stroma) fluxes of 36[Cl] chloride and 22[Na] sodium ions across the CBE were measured under short-circuited conditions. The effect of 0.1 mM of furosemide and bumetanide on the chloride transport were studied. RESULTS: The potential difference (PD), the resistance and the short circuit current (SCC) across the isolated bovine ciliary body were found to be 0.20+/-0.01 mV (aqueous negative), 75+/-1 omegacm2 and -2.70+/-0.17 microAcm(-2) (mean+/-SEM, n=50) respectively. A statistically significant net inward chloride ion flux of 1.12+/-0.41 microEq h(-1)cm(-2) (p < 0.01) was found (n=15). The net chloride transport was abolished when 0.1 mM furosemide (82% inhibition) and 0.1 mM bumetanide (100% inhibition) were applied bilateral. No significant net sodium ion flux was detected. CONCLUSIONS: Electrolyte and fluid transport across the bovine CBE may be via a bumetanide and furosemide-sensitive chloride transport mechanism. The Na-K-2Cl cotransporter plays a significant role in the trans-CBE chloride transport. The net chloride flux/current was about 12 times higher than the measured SCC, suggesting that the chloride ion transport may be coupled to other ion species. PMID- 9746438 TI - Study of regional deformation of the optic nerve head using scanning laser tomography. AB - PURPOSE: Previous studies have suggested that IOP-induced deformation of the optic nerve head (ONH) at the level of the lamina cribrosa may contribute to axonal damage in glaucomatous optic neuropathy. Our purpose was to introduce a novel enucleated eye model for characterizing acute IOP-induced changes in ONH topography, and to develop improved analytical methods for detection of regional topographic change in the ONH. METHODS: Using a specially designed experimental apparatus, enucleated human eyes were progressively pressurized to 5, 15, 30, and 50 mmHg. Seven topographic images of the optic disc were taken at each pressure by a scanning laser tomographer (Heidelberg Retina Tomograph-HRT). The dependence of ONH topography on IOP was quantified for the entire nerve using standard HRT indices of ONH topographic change. The supero-inferior and nasal-temporal hemifields were also analyzed. A new method of analysis was developed which computes the location of the point of maximum slope within a 10 degrees sector of the ONH, as well as the magnitude of this slope. This method, termed "Inflection Point Analysis," was designed to be robust to the potential artefacts of image translation, reference plane location, and the subjective determination of ONH limits. RESULTS: The results of three eyes are presented to illustrate the techniques. In our enucleated eye model, average ONH depth progressively increased with IOP, showing a maximum average posterior displacement of 36 microm as IOP was changed from 5 to 50 mmHg. Significant regional variability in ONH displacement was observed, which both Inflection Point Analysis and standard HRT parameters were able to detect. Inflection point analysis showed several advantages over standard HRT parameters: it was insensitive to artefacts due to tilt, was able to objectively delineate the boundary between the optic cup and neuroretinal rim, and was able to sensitively track changes in the location of this margin. CONCLUSIONS: Scanning laser tomography is capable of detecting regional variation in the deformation of the ONH in response to acute changes in IOP. Our enucleated eye model and Inflection Point Analysis are promising tools for basic studies of ONH deformation in response to IOP. More extensive studies of both enucleated and in vivo eyes are required to determine the potential of Inflection Point Analysis for studying and tracking the progression of glaucomatous optic neuropathy. PMID- 9746439 TI - Absence of p53 delays apoptotic photoreceptor cell death in the rds mouse. AB - PURPOSE: This study was aimed at determining whether or not apoptotic photoreceptor cell death in a mouse model of inherited retinal degeneration is p53 dependent. METHODS: A colony of p53-deficient rds mice were obtained by crossing homozygous rds mice with animals homozygous for a targeted disruption of the p53 gene and genotyping the offspring of the F1 cross. Both parental strains were on a BALB/c background. Age matched p53-deficient rds mice and controls (p53 deficient, rds and BALB/c mice), were sacrificed from day 1 to day 58 after birth. Eyes were paraffin-embedded and a modified terminal dUTP nick-end labeling (TUNEL) technique was used to detect the number of cells displaying DNA fragmentation within the sectioned retina. Eyes were also resin-embedded for semi thin and ultra-thin sectioning. RESULTS: The peak in photoreceptor apoptosis, which occurs at 16 days in the rds mouse, was delayed by 3 days in p53-deficient rds mice. In addition, there was also a delay in the loss of photoreceptor cells between 16 and 26 days. However, absence of p53 did not prevent retinal degeneration in the rds mouse. The number of photoreceptor cells in p53-deficient rds mice at 35 days was very similar to that in the controls. CONCLUSIONS: We have demonstrated that absence of p53 delays but does not prevent photoreceptor cell loss in the rds mouse. Our results provide evidence for plasticity in the mechanism by which apoptosis proceeds in retinal degeneration. PMID- 9746440 TI - The effect of elevated extracellular glucose on migration, adhesion and proliferation of SV40 transformed human corneal epithelial cells. AB - PURPOSE: To examine the effect of elevated extracellular glucose, thus simulating diabetes, on migration, adhesion and proliferation of SV40 transformed human corneal epithelial (HCE) cells. METHODS: HCE cells were maintained in serum supplemented media containing 5 mM, 17.5 mM or 38 mM D-glucose. Cell migration was determined using Blind well chambers fitted with fibronectin/collagen I coated filters. In adhesion experiments, cells were allowed to adhere to extracellular matrix protein-coated wells for 90 min at 37 degrees C. Non adherent cells were removed by washing, then the fluorochrome calcein-AM was added to quantitate the number of attached cells. Proliferation was determined by plating the cells at low density, then quantitating viable cells with calcein-AM 5 to 7 days later. RESULTS: Raising extracellular glucose from 5 mM to 17.5 mM significantly increased cell migration by 42%. When glucose was further raised to 38 mM, migration was not significantly different from that in 5 mM glucose. Adhesion to fibronectin and collagen I (but not IV) was significantly increased (62% and 32% respectively) when cells were cultured in 17.5 mM glucose. Similarly, proliferation was increased by 44%. Adhesion and proliferation tended to be decreased at 38 mM compared to 17.5 mM glucose, but not significantly so. In the presence of 5 mM glucose and mannitol (12.5 mM or 33 mM), neither migration, adhesion nor proliferation were significantly different from that in 5 mM glucose alone. CONCLUSION: Elevated extracellular glucose modulates migration, adhesion and proliferation of HCE cells. The effects are dependent on the concentration of glucose and are not due to changes in osmolality since mannitol failed to produce similar results. Our in vitro findings suggest that high glucose effects may directly contribute to the etiology of impaired corneal wound healing in diabetes. PMID- 9746441 TI - OPC-15161 suppresses the proliferation of Tenon's capsule fibroblasts and the production of type I collagen and fibronectin stimulated by TGF-beta1 in vitro. AB - PURPOSE: We investigated the effects of OPC-15161 on the growth of cultured human Tenon's capsule fibroblasts (TCFs), as well as on the production of type I procollagen, fibronectin, and laminin. These effects were examined in the presence or absence of TGF-beta1. METHODS: Cell proliferation was assayed by counting cell number and assay of DNA synthesis. Cytotoxicity was determined by the MTT method. Matrix components were assayed by enzyme immunoassay of material in the medium and in the cell lysate with or without OPC-15161. Total protein content was determined. Cellular ultrastructure was also evaluated. RESULTS: Treatment with OPC-15161 (up to 100.0 microg ml(-1)) significantly reduced the proliferation and DNA synthesis of TCFs. No significant decrease in MTT values was observed in confluent TCF cultures with OPC-15161 (up to 100.0 microg ml( 1)). TGF-beta1 enhanced the TCF production of procollagen I and fibronectin. OPC 15161 significantly decreased the procollagen I content in both the medium, in the cell lysate of TGF-beta1-stimulated cells, and fibronectin content in the lysate. OPC-15161 did not affect the laminin or total protein content, either with or without TGF-beta1. No ultrastructural evidence of cytotoxicity was observed. CONCLUSIONS: OPC-15161 inhibited the proliferation of TCFs, and reduced their production of procollagen I and fibronectin in the presence of TGF-beta1 without evidence of cytotoxicity. OPC-15161 may be useful in inhibiting the excessive fibrosis produced in the wound in response to filtering surgery. PMID- 9746442 TI - The effect of genetic deficiency of adhesion molecules on the course of endotoxin induced uveitis. AB - PURPOSE: Multiple adhesion molecules of the selectin, integrin, and immunoglobulin-like families are involved in the migration of leukocytes out of the bloodstream into inflamed tissues. This study addresses the question of which adhesion molecules are specifically involved in endotoxin-induced uveitis. METHODS: Mice genetically deficient in p-selectin, ICAM-1, beta2-integrin, or controls received intravitreal injections of endotoxin. Eyes were harvested 24 h later and inflammation was evaluated by histologic and immunohistochemical assays of infiltrating cells. RESULTS: Mice lacking either P-selectin or beta2-integrin had less inflammation than controls (median cells/section: 64 for P-selectin knockout vs 130 for controls, p=0.02, n=17 per group: 244 for beta2-integrin knockouts, n=14, vs 355 for controls, n=17, p=0.05). Neither gene deletion significantly changed the ratio of infiltrating neutrophils to macrophages. ICAM 1 knockouts tended to have fewer infiltrating cells (median 22 cells/section) compared to controls (median 132 cells/section), but this difference was not statistically significant (p=0.25, n=9 per group). CONCLUSIONS: P-selectin, beta2 integrin, and possibly ICAM-1 are involved in the ocular inflammatory response to endotoxin. The lack of complete inhibition of leukocyte infiltration with the complete loss of each adhesion molecule is in accord with the notion that alternative adhesion molecules also participate in this process. PMID- 9746443 TI - Presenilin expression in the ocular lens. AB - PURPOSE: Mutations in the presenilin (PS) proteins account for the majority of early onset Alzheimer's disease (AD) cases, apparently by influencing the cleavage of the Alzheimer's disease protein (betaAPP) to form beta-amyloid (Abeta), the major component of plaques in the brains of AD patients. We reported previously that AD proteins are expressed in mammalian lenses, and that betaAPP and Abeta increased in the epithelium and outer cortex of lenses subjected to oxidative stress. This increase paralleled the increase in AP1 DNA binding activity, which has been shown to accompany proliferative oxidative stress responses. Both cataract and AD have been closely linked with oxidative stress; further, both AD and cataract occur in a majority of Down Syndrome individuals. Here we investigate the expression and post-translational processing of PS proteins in the ocular lens. METHODS: In situ hybridization, immuohistochemical detection and immunoblot assays were used to localize mRNA and proteins expression products and determine the approximate molecular weights of the resulting proteins in ocular tissue samples. RESULTS: We report here that PS protein and mRNA are expressed in lenses, and additionally in the cornea, and are proteolytically processed in a manner similar to that demonstrated in brain tissue. PS proteins and mRNAs were localized to the lens epithelium and outer fibers. This pattern agrees with the localization demonstrated by others for mammalian Notch-like receptor proteins. PS and Notch proteins occur together in developmentally regulated cascades of gene expression found in diverse biological systems. CONCLUSIONS: PS expression, together with betaAPP and Abeta proteins, all associated with age-related degenerative disease, are expressed in lens and might contribute to cataractogenesis. PMID- 9746444 TI - Antidepressant effect of transdermal nicotine patches in nonsmoking patients with major depression. AB - BACKGROUND: A high frequency of cigarette smoking has been reported among individuals with major depression. In a previous study, transdermal nicotine produced short-term improvement in the mood of depressed patients. This study was undertaken to determine the effects of 4 days' administration of transdermal nicotine on mood in nonsmoking patients with major depression. METHOD: The effects of nicotine patches (17.5 mg) on 12 nonmedicated outpatients with major depression (DSM-III-R) were studied for 4 continuous days. Patients had to score 18 points or more on the Hamilton Rating Scale for Depression (HAM-D) (21 items) for admission into the study. The HAM-D (10 items), a visual analog scale, and a side effects scale were scored daily during the trial (baseline, 4 days of nicotine patches, and 4 days of follow-up). RESULTS: Two patients dropped out of the study owing to nausea and vomiting. Results of the visual analog scale and HAM-D (10 items) showed a significant (p < .01) improvement in depression after the second day of nicotine patches. Patients relapsed 3 or 4 days after the last nicotine dose. The side effects observed were an increase in saliva production, nausea, loss of appetite, and mild insomnia. CONCLUSION: Nicotine patches produced short-term improvement of depression with minor side effects. Because of nicotine's high risk to health, nicotine patches are not recommended for clinical use in depression. Analogue drugs may be developed in the future that may help improve depression without the risk of other major health problems. PMID- 9746445 TI - Treatment of recent trauma survivors with benzodiazepines: a prospective study. AB - BACKGROUND: Most types of psychotropic drugs have been tried in the treatment of chronic posttraumatic stress disorder (PTSD), but have yielded limited results. Theory and retrospective research predict that early treatment may be more efficacious. Specifically, high-potency benzodiazepines have been recommended for the treatment of acute responses to trauma and for prevention of PTSD. This study prospectively evaluates the effect of early administration of benzodiazepines on the course of PTSD and PTSD symptoms. METHOD: Thirteen trauma survivors (the benzodiazepine group) were treated within 6.7 +/- 5.8 days after the trauma (range, 2-18) with either clonazepam (N = 10, 2.7 +/- 0.8 mg/day) or alprazolam (N = 3, 2.5 mg/day). Thirteen other trauma survivors, pair-matched with subjects in the active treatment group for gender and symptom severity in the first week after the trauma, constitute the control group. Both groups were reevaluated 1 and 6 months after the trauma for PTSD symptoms (Horowitz Impact of Event Scale; Mississippi Rating Scale for Combat-Related PTSD-civilian trauma version), PTSD status (Clinician Administered PTSD Scale), state anxiety, depression, and resting heart rate. RESULTS: Subjects in the benzodiazepine group did not differ from controls in 1-month and 6-month PTSD and anxiety scores. Repeated measures ANOVA showed no group or group-by-time effect on psychometric measures. A trend toward group-by-time interaction in resting heart rate was noted (progressive decrease in the benzodiazepine group). Nine benzodiazepine subjects and 3 controls met PTSD diagnostic criteria 6 months after the trauma. CONCLUSION: Contrary to expectations, the early administration of benzodiazepines to trauma survivors with high levels of initial distress did not have a salient beneficial effect on the course of their illness, while reducing physiologic expression of arousal. PMID- 9746446 TI - Risperidone as an adjunct to clozapine therapy in chronic schizophrenics. AB - BACKGROUND: Some treatment-resistant schizophrenic patients improve enough to remain out of the hospital but continue to have significant positive or negative symptoms. METHOD: The goal of this study was to assess the safety and potential efficacy of risperidone as an adjunct for schizophrenic patients treated with clozapine. In an open 4-week trial involving 12 DSM-III-R-diagnosed patients, the addition of risperidone to clozapine was well tolerated and did not affect serum clozapine concentrations significantly. RESULTS: Total Brief Psychiatric Rating Scale (BPRS) scores and subscales measuring positive symptoms, negative symptoms, and depressive symptoms were significantly reduced from baseline. Ten of 12 participants had a 20% or greater reduction in the total BPRS score. CONCLUSION: In this open trial, the addition of risperidone to clozapine was well tolerated and produced significant reduction of symptoms, suggesting that this may be a useful clinical approach. Because this was an open trial, the improvement we observed must be replicated in a controlled trial. PMID- 9746447 TI - Fluoxetine in the treatment of anger: an open clinical trial. AB - BACKGROUND: Intense anger, easy irritability, and low frustration tolerance are frequently encountered in clinical practice and may occur as part of the constellation of symptoms associated with a number of Axis I and Axis II diagnoses. Recent studies have suggested the efficacy of fluoxetine in treating anger in depressed and borderline patients. The present study examines the treatment of anger as a target symptom regardless of accompanying psychopathology. METHOD: In this preliminary trial, 11 subjects experiencing intense anger were treated in an open-label fashion with fluoxetine over an 8 week period. Anger was measured by a clinician-administered Clinical Global Impressions-Severity (CGI-S) scale and a self-report anger inventory, the Multidimensional Anger Inventory (MAI). The Hamilton Rating Scale for Depression was administered at baseline and final visits. RESULTS: All 11 enrolled patients completed the study, and all patients demonstrated clinical improvement as measured by both the CGI-S and the MAI. Nine patients were rated as responders and 2 as nonresponders according to prospectively established criteria. Fluoxetine was well tolerated and showed rapid onset of action. CONCLUSION: This study is limited by small sample size and nonblinded, open-label design, but nonetheless suggests that fluoxetine may be useful in moderating anger as it occurs as part of the symptomatology of a number of Axis I and Axis II disorders. Controlled studies are warranted. PMID- 9746448 TI - Panic attacks with psychotic features. AB - BACKGROUND: Anxiety disorders are among the most frequently diagnosed group of psychiatric disorders in the general population. Although anxiety disorders are often comorbid with depression and personality disorders, they rarely culminate in psychosis. METHOD: Having observed psychosis in the course of a severe panic attack, the authors prospectively identified four patients who experienced panic attacks with co-occurring psychosis. All met the DSM-IV criteria for panic disorder. Distinctive features of their clinical presentation, pharmacotherapy, and follow-up were recorded. RESULTS: Three patients had a history of panic disorder, and one had a history of generalized anxiety disorder. In all cases, psychosis (auditory hallucinations or delusions) originated in the course of a severe panic attack. Psychotic symptoms occurred only during panic attacks; however, these could occur up to 10 to 15 times a day. In all four patients, psychotic symptoms resolved after a brief time either spontaneously or with benzodiazepine/SSRI treatment. None of the patients required neuroleptic treatment. CONCLUSION: The cases suggest that psychosis may develop in the course of a severe panic attack in patients with panic disorder, as was reported previously for patients with obsessive-compulsive disorder and posttraumatic stress disorder. Distinguishing panic attacks with psychotic features from other psychotic disorders is clinically important since antipsychotic medication treatment for these psychotic symptoms is not indicated. Further research on the prevalence of psychotic symptoms in the anxiety disorders and the pathophysiology of this phenomenon is required to clarify the relationship between the anxiety disorders and psychosis. PMID- 9746450 TI - Suicidal risk during controlled clinical investigations of fluvoxamine. AB - BACKGROUND: Suicide is a serious risk factor in major depressive disorder. Paradoxical emergence of suicidal ideation or behavior during antidepressant treatment has been reported in isolated cases. An evaluation was undertaken to assess the risk of suicidality during treatment with fluvoxamine, a serotonin selective reuptake inhibitor. METHOD: Meta-analyses were conducted on pooled data from double-blind, randomized, placebo-controlled, parallel-group clinical trials. The primary outcome measure was the suicide item of the Hamilton Rating Scale for Depression. Tests for emergence of substantial suicidal ideation and improvement or worsening in suicidal ideation were performed using the Mantel Haenszel adjusted incidence difference. The Breslow-Day test was used to test for lack of homogeneity across trials. Secondary analysis, which consisted of Pearson's chi-square test, was used to confirm the Mantel-Haenszel result. RESULTS: In comparison to placebo, fluvoxamine was associated with significantly greater improvement in suicidal ideation (p = .01) and significantly less worsening of suicidal ideation (p < .01). No differences were found in the emergence of substantial suicidal ideation. CONCLUSION: These findings demonstrate that fluvoxamine is not associated with an increased risk of emergence of substantial suicidal thoughts among depressed patients. On the contrary, the results are suggestive of a protective effect of fluvoxamine upon the risk of suicidal ideation. PMID- 9746449 TI - Neuropsychiatric aspects of Sydenham's chorea: a comprehensive review. AB - BACKGROUND: The recent demonstration of an etiologic role for Sydenham's chorea in obsessive-compulsive disorder has once again brought this disorder to psychiatric attention. Despite its traditional importance to psychiatry, there has not been a comprehensive review of Sydenham's chorea published for decades. METHOD: I utilized the Index Medicus and its predecessors, the Quarterly Cumulative Index Medicus and the Cumulative Index Medicus, to search for every English-language article on Sydenham's chorea written during the past 115 years and then read and compiled the articles and their pertinent references. RESULTS: Sydenham's chorea is a symptom of rheumatic fever and results from an autoimmune attack on the CNS. In addition to chorea, the acute attack is almost always characterized by psychiatric symptoms such as irritability, obsessions and compulsions, tics, and psychotic symptoms. Acutely, treatment involves valproic acid or haloperidol; steroids may also be appropriate. Chronic treatment with penicillin is required to prevent future attacks. In adult years, the neuropsychiatric sequelae of Sydenham's chorea include chorea gravidarum, some cases of obsessive-compulsive disorder, and possibly, certain cases of Tourette's syndrome and of schizophrenia. CONCLUSION: Sydenham's chorea is best considered a neuropsychiatric disorder, and most patients will benefit from psychiatric treatment. Certain cases of obsessive-compulsive disorder, and perhaps other adult psychiatric disorders as well, may benefit from approaches similar to those used for Sydenham's chorea. PMID- 9746451 TI - Generalized edema with risperidone: divalproex sodium treatment. PMID- 9746453 TI - Breast pain associated with venlafaxine. PMID- 9746452 TI - Possible risks of long-term zinc supplementation. PMID- 9746454 TI - Treatment of obsessive-compulsive cutting behavior with naltrexone. PMID- 9746455 TI - Cocaine-related obsessive homicidal ideation. PMID- 9746456 TI - Choosing among old and new antipsychotics. PMID- 9746457 TI - Augmented renal sympathetic nerve activity by central command during overground locomotion in decerebrate cats. AB - We examined whether the cerebrum is essential for producing the rapid autonomic adjustment at the onset of spontaneous overground locomotion. Renal sympathetic nerve activity (RSNA), mean arterial pressure (MAP), heart rate (HR), and electromyogram of the forelimb triceps brachialis were measured when freely moving, decerebrate cats spontaneously produced overground locomotion, supporting body weight. Decerebration was performed at the level of the precollicular premammillary body. RSNA increased 95 +/- 14 impulses/s (68 +/- 10% of baseline value) at the onset of spontaneous locomotion, which was followed by rises in MAP and HR (7 +/- 1 mmHg and 18 +/- 2 beats/min, respectively). Concomitantly with the MAP rise, RSNA declined toward control values and then increased again during the subsequent period of locomotion. The same rapid increase in RSNA at the onset of locomotion was observed after sinoaortic denervation and vagotomy. It is concluded that some central site(s), other than the cerebrum and the rostral part of the diencephalon, can generate the centrally induced autonomic adjustment at the onset of spontaneous overground locomotion, which is independent of arterial baroreceptor and vagal afferents. PMID- 9746458 TI - Induction and cDNA sequence of inducible nitric oxide synthase from canine aortic smooth muscle cells. AB - An inducible isoform of nitric oxide synthase (type II, iNOS) is expressed in cardiac and vascular smooth muscle in response to inflammatory cytokines. The dog is an important large animal used for cardiovascular research including effects of exercise, heart failure, and allograft rejection. However, molecular probes for iNOS developed in other mammals have not been reliable for the study of iNOS induction in canine vascular smooth muscle. Experiments were designed to develop a molecular probe for canine iNOS. Smooth muscle cells were isolated from canine aortas. The cells (passages 3-10) were incubated for 1, 3, 6, 12, 24, 48, or 72 h in the absence and presence of Escherichia coli lipopolysaccharide (LPS) to induce iNOS. Total RNA was isolated from the cells using standard techniques. RT PCR with primers against conserved regions of all known iNOS enzyme was used to clone the iNOS cDNA. RT-PCR showed a single band only from cells treated with LPS. Cloned cDNA from cultured canine aortic smooth muscle cells has 84% homology to human, 81% to rat, and 81% to mouse iNOS gene. Identification of the cDNA for canine iNOS will be useful in the study of differential, transcriptional regulation of inducible (type II) compared with constitutive endothelial (type III) NOS in canine studies of allograft rejection and cardiovascular disease. PMID- 9746459 TI - Cardiovascular, endocrine, and body fluid-electrolyte responses to salt loading in mRen-2 transgenic rats. AB - We previously demonstrated that mRen-2 transgenic [Tg(+)] rats are sensitive to chronic high NaCl intake, showing increased arterial pressure and vasopressin (VP) secretion. In this study, we determined the effect of a chronic osmotic challenge, 4 days of drinking 2% NaCl, on direct arterial blood pressure, heart rate, fluid-electrolyte balance, circadian rhythm of mean arterial pressure (MAP), and changes in plasma VP and catecholamines. Under baseline conditions, male Tg(+) rats showed a significant shift in the peak in circadian MAP into the light portion of the day-night cycle. Substitution of 2% NaCl for drinking water caused a rapid increase in MAP, 20 +/- 5 mmHg in Tg(+) rats within 6 h. Whereas the amplitude of circadian MAP fluctuations increased in salt-loaded Tg(+) rats, there was no significant change in the circadian timing of peak MAP with salt loading. Tg(+) rats showed exaggerated osmotic-induced increases in plasma VP, norepinephrine (NE), and epinephrine (Epi) compared with Tg(-) rats. Plasma NE and Epi were increased two- and fourfold, respectively, in the hypertensive rats with no significant change in the Tg(-) rats. Intravenous administration of a VP antagonist did not alter arterial pressure in either Tg(+) or Tg(-) rats. Tg(+) and Tg(-) rats showed a positive sodium balance with no significant difference observed between the groups. Tg(+) rats showed a significant increase in salt consumption, plasma sodium, osmolality, and hematocrit, accompanied by a negative water balance. We conclude that Tg(+) rats are sensitive to acute and chronic osmotic stimuli in terms of blood pressure, fluid-electrolyte balance, and plasma VP and catecholamines. Whereas elevated plasma VP does not contribute to the hypertensive response, increased sympathetic drive may mediate the salt-induced blood pressure changes in this model. PMID- 9746460 TI - Neutrophil-dependent augmentation of PAF-induced vasoconstriction and albumin flux in coronary arterioles. AB - Platelet-activating factor (PAF) has been implicated in the pathogenesis of ischemic heart disease, reperfusion injury, and inflammatory reactions. Although neutrophils have been shown to primarily mediate PAF-induced microvascular dysfunction, the vasoactive effect of PAF and its neutrophil-dependent mechanism have not been directly and systematically studied in coronary resistance vessels. Therefore, the aim of this study was to examine the effects of PAF on coronary arteriolar function and neutrophil dynamics using an isolated and perfused microvessel preparation. Topical application of PAF to the vessels induced a dose dependent decrease in the diameter but an increase in the apparent permeability coefficient of albumin. Disruption of the endothelium abolished the vasomotor response to PAF, and perfusion of neutrophils significantly augmented PAF-induced changes in vasomotor tone and permeability. Furthermore, the interaction between neutrophils and the endothelium was studied in the intact perfused coronary arterioles. Under control conditions, there were no adherent neutrophils observed in the vessels at varied intraluminal flow velocities. However, administration of PAF caused neutrophil adhesion to the endothelium of coronary arterioles at low flow velocities. Western blot analysis indicated that PAF upregulated the expression of intercellular adhesion molecule-1 in cultured coronary microvascular endothelial cells. Taken together, the results suggest that 1) PAF induces vasoconstriction and hyperpermeability in coronary arterioles via an endothelium-dependent and neutrophil-mediated mechanism, and 2) PAF is able to stimulate neutrophil adhesion in coronary arterioles under a condition of low flow rate. PMID- 9746461 TI - Nitric oxide-independent activation of soluble guanylyl cyclase contributes to endotoxin shock in rats. AB - We investigated whether a complete inhibition of nitric oxide (NO) formation caused by bacterial endotoxin (lipopolysaccharide, LPS) in vivo prevents the hypotension and restores the vascular hyporeactivity to normal in vivo and ex vivo. The combination of dexamethasone (Dex; 3 mg/kg at 30 min before LPS) plus aminoguanidine (AG; 15 mg/kg at 2 h after LPS) inhibited the overproduction of nitrate (an indicator of NO) in the plasma and aortic smooth muscle and also prevented the development of the delayed hypotension in rats treated with LPS for 6 h. However, the vascular hyporeactivity to norepinephrine (NE) was only partially improved either in vivo or ex vivo in endotoxemic rats treated with Dex plus AG. Pretreatment of aortic rings with Nomega-nitro-L-arginine methyl ester (L-NAME) or 1H-[1,2, 4]oxidazolo[4,3-a]quinoxalin-1-one (ODQ) enhanced the contraction to NE in rings obtained from LPS-treated rats, but not in those from Dex plus AG-treated endotoxemic rats. Methylene blue, an inhibitor of soluble guanylyl cyclase (GC), completely restored contractions to NE and aortic cGMP levels to normal either in LPS-treated rats or in Dex plus AG-treated endotoxemic rats, whereas the cGMP level was partially inhibited by ODQ in LPS-treated rats only. These results suggest that non-NO mediator(s) also activates soluble GC during endotoxemia. Interestingly, we found that in the presence of tetraethylammonium (an inhibitor of K+ channels) plus L-NAME or charybdotoxin [a specific inhibitor of large-conductance Ca2+-activated K+ (KCa) channels] plus ODQ, the vascular hyporeactivity to NE in the LPS-treated group was also completely restored to normal. In addition, in the presence of L-NAME or ODQ, the vascular hyporeactivity to high K+ was abolished in rings from the LPS-treated group. These results suggest that LPS causes the production of other mediator(s), in addition to NO, which also stimulates soluble GC (i.e., increases the formation of cGMP) and then activates the large-conductance KCa channels in the vascular smooth muscle causing vascular hyporeactivity. PMID- 9746462 TI - Regional differences in effects of 4-aminopyridine within the sinoatrial node. AB - 4-Aminopyridine (4-AP)-sensitive transient outward current (Ito) has been observed in the sinoatrial node, but its role is unknown. The effect of block of Ito by 5 mM 4-AP on small ball-like tissue preparations (diameter approximately 0.3-0.4 mm) from different regions of the rabbit sinoatrial node has been investigated. 4-AP elevated the plateau, prolonged the action potential, and decreased the maximum diastolic potential. Effects were greater in tissue from the periphery of the node than from the center. In peripheral tissue, 4-AP abolished the action potential notch, if present. 4-AP slowed pacemaker activity of peripheral tissue but accelerated that of central tissue. Differences in the response to 4-AP were also observed between tissue from more superior and inferior regions of the node. In the intact sinoatrial node, 4-AP resulted in a shift of the leading pacemaker site consistent with the regional differences in the response to 4-AP. It is concluded that 4-AP-sensitive outward current plays a major role in action potential repolarization and pacemaker activity in the sinoatrial node and that its role varies regionally. PMID- 9746463 TI - Reflex vascular responses in the anesthetized dog to large rapid changes in carotid sinus pressure. AB - This study examined reflex vascular responses to large rapid increases and decreases in carotid sinus pressure to determine whether delayed or inappropriate vascular responses might be obtained that, if they occurred in people, could lead to hypotension during exposure to rapidly alternating gravitational forces. In chloralose-anesthetized open-chest dogs, a perfusion circuit controlled carotid sinus and thoracic aortic pressures and blood flows to both the vascularly isolated abdominal circulation and a hindlimb (perfusion pressure changes denoted resistance). When carotid pressure was increased and decreased over the range of 60-180 mmHg, the resulting reflex vasodilatation occurred significantly more rapidly than the vasoconstriction (P < 0.001). In the abdominal vascular bed, time constants for vasodilatation and vasoconstriction were 4.2 +/- 0.5 and 7.5 +/- 1.0 s, respectively. Decreases in carotid pressure in pulses of 10-s duration or less failed to elicit maximal vasoconstriction, whereas increases in carotid pressure lasting as little as 5 s did elicit maximal vasodilatation. "Square wave" alternations in carotid pressure with periods of 10 s or less (5 s high, 5 s low) resulted in attenuation of the vasoconstriction, and at a 4-s period, both vascular beds remained almost maximally vasodilated throughout. The failure of vascular resistance to follow carotid pressure changes was not due to a failure of the response of sympathetic efferent activity, since the time constants for the reduction and increase in discharge were much shorter at 0.56 +/- 0.13 and 0.43 +/- 0.10 s, respectively. These results indicate that rapid changes in carotid pressure could result in inappropriate vasodilatation and hypotension and might, in some circumstances, such as in pilots flying high-performance aircraft, predispose to syncope. PMID- 9746464 TI - Role of central circulatory factors in the fat-free mass-maximal aerobic capacity relation across age. AB - Fat-free mass (FFM) (primarily skeletal muscle mass) is related to maximal aerobic capacity among healthy humans across the adult age range. The basis for this physiological association is assumed to be a direct relation between skeletal muscle mass and its capacity to consume oxygen. We tested the alternative hypothesis that FFM exerts its influence on maximal aerobic capacity in part via an association with central circulatory function. To do so, we analyzed data from 103 healthy sedentary adults aged 18-75 yr. FFM was strongly and positively related to maximal oxygen consumption (r = 0.80, P < 0. 001). FFM was also strongly and positively related to supine resting levels of blood volume (r = 0.79, P < 0.001) and stroke volume (r = 0.75, P < 0.001). Statistically controlling for the collective influences of blood volume and stroke volume abolished the tight relation between FFM and maximal oxygen consumption (r = 0.12, not significant). These results indicate that 1) FFM may be an important physiological determinant of blood volume and stroke volume among healthy sedentary adult humans of varying age; and 2) this relation between FFM and central circulatory function appears to represent the primary physiological basis for the strong association between FFM and maximal aerobic capacity in this population. Our findings suggest that sarcopenia (loss of skeletal muscle mass with aging) may contribute to the age-related decline in maximal aerobic capacity primarily via reductions in blood volume and stroke volume rather than a direct effect on the oxygen-consuming potential of muscle per se. PMID- 9746465 TI - Effects of red and white wine on endothelium-dependent vasorelaxation of rat aorta and human coronary arteries. AB - Beneficial effects of wine on myocardial infarction mortality may be because of its vasodilatory properties. This study investigated whether the vasodilatory activity involves the endothelium and is specific for certain wines. Effects of different red and white wines and phenolic grape ingredients on vascular tension and cGMP content were studied in human coronary arteries and rat aortic rings in vitro. Only French and Italian red wines produced "en barrique" (Bordeaux, Chateauneuf du Pape, Barolo) (1:1,000, vol/vol), quercetin (1-100 microM), and tannic acid (1-100 microgram/ml) decreased tension of precontracted vascular rings and increased vascular cGMP content (both P < 0.001). The effects were abolished after endothelial denudation and reversible by nitric oxide synthase inhibition. Red wines not produced en barrique (Valpolicella, Ahr Spatburgunder), white wines (en barrique-produced Rioja, Chardonnay, Mosel-Riesling), and ethanol did not affect vascular tension or cGMP content. Thus endothelium-dependent vasodilatory effects appear to be specific for red barrique wines, possibly because of their high content of phenolic substances. Divergent effects of wines indicate that a general view on the effects of wine and alcoholic beverages is not warranted. PMID- 9746466 TI - 31P NMR studies of creatine kinase flux in M-creatine kinase-deficient mouse heart. AB - Hearts of wild-type and cytosolic muscle creatine kinase (M-CK)-knockout mice were perfused with Krebs-Henseleit buffer containing 10 mM glucose and 5 mM pyruvate and studied during pacing at 400 and 600 beats/min and during K+ arrest. Phosphocreatine (PCr) and ATP concentrations in M-CK-deficient hearts were not significantly different from those in wild-type hearts. With the use of 31P NMR saturation transfer, the flux mediated predominantly by mitochondrial creatine kinase (Mi-CK) was clearly detected in M-CK-deficient hearts. Mi-CK flux was 4.8 +/- 0.6 and 4.5 +/- 0.6 mM/s during pacing at 400 and 600 beats/min, respectively, and was 3. 5 +/- 0.4 mM/s during cardiac arrest. In control hearts total CK flux was 7.8 +/- 1.1 and 6.6 +/- 1.3 mM/s during pacing at 400 and 600 beats/min, respectively, and decreased to 3.8 +/- 0.5 mM/s during arrest. It is suggested that the relative contribution of Mi-CK to the total NMR-measured CK flux in the wild-type heart is higher than that of the homodimeric M-CK isoform (MM-CK). PMID- 9746467 TI - Amino-terminal proBNP in ovine plasma: evidence for enhanced secretion in response to cardiac overload. AB - We have recently identified a novel amino-terminal fragment of pro-brain natriuretic peptide (NT-proBNP) in the circulation of humans, the concentration of which increases progressively as the left ventricle fails. To clarify the origins of NT-proBNP in experimental animals, we have developed an RIA for NT proBNP based on residues 52-71 of ovine proBNP-(1-103) and used it to study cardiac processing, secretion, and metabolism of BNP in sheep with cardiac overload induced by coronary artery ligation (CAL) or rapid left ventricular pacing (rLVP). The concentration of NT-proBNP in left atrial plasma extracts drawn from normal control sheep was threefold that of mature BNP. Size-exclusion and reverse-phase HPLC analyses of plasma extracts coupled to RIA revealed a single peak of immunoreactive (ir) NT-proBNP [ approximately 8,000 relative molecular weight (Mr)], quite distinct from a single peak of ir-mature BNP ( approximately 3,000 Mr). In contrast, ovine cardiac tissue contained only a single immunoreactive peak of high-molecular-weight BNP ( approximately 11,000 Mr), consistent in size with proBNP-(1-103). Sampling from the cardiac coronary sinus in normal control sheep (n = 5) and sheep with CAL (n = 5) revealed that the molar ratio of NT-proBNP to mature BNP was similar. There was a significant gradient of both mature and NT-proBNP across the heart in normal sheep, whereas after CAL the gradient was significant for mature BNP only. In both forms of cardiac overload (CAL and rLVP), left atrial plasma levels of NT-proBNP were significantly increased above normal levels, in contrast with mature BNP levels, which were raised only in the rLVP group of animals. Blockade of natriuretic peptide metabolism in sheep with heart failure (induced by rLVP) raised mature BNP levels threefold but did not affect levels of NT-proBNP. In conclusion, these studies show that NT-proBNP is formed from proBNP stores during secretion and, compared with mature BNP, accumulates in plasma because metabolism of NT-proBNP appears to differ from that of mature BNP. Although its function, if any, remains unclear, plasma NT-proBNP may prove to be a sensitive marker of cardiac overload and/or decompensation. PMID- 9746469 TI - Regression of LV hypertrophy with captopril normalizes membrane currents in rabbits. AB - Recent studies indicate that regression of left ventricular hypertrophy (LVH) normalizes the in situ electrophysiological abnormalities of the left ventricle. This study was designed to determine whether regression of LVH also normalizes the abnormalities of individual membrane currents. LVH was induced in rabbits by renal artery banding. Single ventricular myocytes from rabbits with LVH at 3 mo after renal artery banding demonstrated increased cell membrane capacitance, prolonged action potential duration, decreased inward rectifier K+ current density, and increased transient outward K+ current density compared with myocytes from age-matched controls. Additional rabbits were randomized at 3 mo after banding to treatment with either vehicle or captopril for an additional 3 mo. Myocytes from LVH rabbits treated with vehicle showed persistent membrane current abnormalities. However, myocytes isolated from LVH rabbits treated with captopril had normal cell membrane capacitance, action potential duration, and membrane current densities. Captopril had no direct effect on membrane currents of either control or LVH myocytes. These data support the hypothesis that the action potential prolongation and membrane current abnormalities of LVH are reversed by regression. Normalization of membrane currents probably explains the reduced vulnerability to ventricular arrhythmia observed in this LVH model after treatment with captopril. PMID- 9746468 TI - ATP-stimulated smooth muscle cell proliferation requires independent ERK and PI3K signaling pathways. AB - Vascular smooth muscle cells respond to the purinergic agonist ATP by increasing intracellular calcium concentration and increasing the rate of cell proliferation. In many cells the extracellular signal-regulated kinase (ERK) cascade plays an important role in cellular proliferation. We have studied the effect of extracellular ATP on ERK activation and cell proliferation. ATP binding to a UTP-sensitive P2Y nucleotide receptor activates ERK1/ERK2 in a time- and dose-dependent manner in coronary artery smooth muscle cells (CASMC). ATP-induced activation of ERK1/ERK2 is dependent on the dual-specificity kinase mitogen activated protein kinase/ERK kinase (i.e., MEK) but independent of phosphatidylinositol 3-kinase (PI3K) activity. We provide evidence that both ERK1/ERK2 and PI3K activities are required for CASMC proliferation. Thus ATP stimulation of CASMC proliferation requires independent activation of both the ERK and PI3K signaling pathways. PMID- 9746470 TI - Are interatrial band myocytes maximally hypertrophied in normal canine hearts? AB - In canine right atrial hypertrophy, the cross-sectional area (Axs) of right atrial myocytes increases, whereas the Axs of the broader interatrial band myocytes does not. In the current study, myocyte reconstructions showed that right atrial myocyte length increased in proportion to Axs in right atrial hypertrophy. On the other hand, mean interatrial band myocyte length in both normal and right atrial hypertrophy dogs was roughly inversely proportional to mean Axs, as expected if interatrial band myocyte volume was constant. Plotting mean Axs vs. myocyte length for individual interatrial band myocytes revealed a distribution whose border defined a maximal volume curve; many myocytes were well beneath that curve. Mononuclear myocytes (generally diploid) were limited by a 65,000-micrometer 3 curve, which many binuclear myocytes (generally tetraploid) surpassed; myocyte ploidy thus constrained myocyte volume. However, because many mononuclear and binuclear myocytes had lower volumes, their failure to hypertrophy cannot be attributed to attainment of the maximal volume possible for their ploidy. PMID- 9746471 TI - Non-NMDA and NMDA receptors in the synaptic pathway between area postrema and nucleus tractus solitarius. AB - Area postrema (AP) modulates cardiovascular function through excitatory projections to neurons in nucleus tractus solitarius (NTS), which also process primary sensory (including cardiovascular-related) input via the solitary tract (TS). The neurotransmitter(s) and their receptors in the AP-NTS pathway have not been fully characterized. We used whole cell recordings in voltage- and current clamp modes in the rat brain stem slice to examine the role of ionotropic glutamatergic receptors and alpha2-adrenergic receptors in the pathway from AP to NTS neurons receiving visceral afferent information via the TS. In neurons voltage clamped at potentials from -100 to +80 mV, AP stimulation (0. 2 Hz) evoked excitatory postsynaptic currents having a fast component blocked by the non-N-methyl-D-aspartate (NMDA) receptor antagonist 1,2,3,4-tetrahydro-6-nitro-2, 3-dioxobenzoquinoxaline-7-sulfonamide (NBQX; 3 microM, n = 7) and a slow component blocked by the NMDA receptor antagonist DL-2-amino-5-phosphonovaleric acid (APV; 50 microM, n = 8). Although NBQX (3 microM, n = 14) abolished AP evoked action potentials, APV (50 microM, n = 9 or 500 microM, n = 6) or yohimbine, (200 nM, n = 5 or 2 microM, n = 10) did not. Thus, although AP stimulation activates both non-NMDA and NMDA receptors on NTS neurons receiving TS input, only non-NMDA receptors are required for synaptic transmission. PMID- 9746472 TI - Increased expression of glomerular AT1 receptors in rats with myocardial infarction. AB - Rats with congestive heart failure demonstrate striking intrarenal vasoconstriction that contributes to reduced renal excretory function. We previously demonstrated that inhibition of angiotensin action reverses intrarenal vasoconstriction in rats 4-6 wk after coronary artery ligation. In the present study we tested the hypothesis that abnormalities in the expression and regulation of glomerular angiotensin receptors contribute to the intrarenal vasoconstriction. Because glomerular angiotensin type 1 (AT1) receptors normally downregulate in response to high local ANG II concentrations, we anticipated that glomerular AT1-receptor expression would be reduced in rats after myocardial infarction (MI). To our surprise, the density of glomerular AT1 receptors was nearly double (97% increase, P < 0.002) that of controls, indicating an acquired abnormality in angiotensin receptor regulation. This was specific for renal glomeruli, because the density of angiotensin receptors on renal vasculature was decreased in rats after MI compared with normal controls. Glomerular AT1-receptor expression was downregulated by an acute pharmacological infusion of ANG II and upregulated by acute angiotensin-converting enzyme inhibition to a similar extent in MI and control rats. Renal cortical mRNA expression showed an increase in the renin mRNA-to-actin ratio and angiotensinogen-to-actin ratio, indicating stimulation of the intrarenal angiotensin system in rats after MI. The data indicate a specific dysregulation of AT1 receptors in glomeruli but not blood vessels after MI. PMID- 9746473 TI - Responses of muscle sympathetic nerve activity to lower body positive pressure. AB - To evaluate the response of vasomotor sympathetic nerve activity to lower body positive pressure (LBPP), muscle sympathetic nerve activity (MSNA) was microneurographically recorded from the tibial nerve in 10 healthy young men, along with hemodynamic variables and echocardiogram, during exposure to incremental LBPP at 10, 20, and 30 mmHg in the supine position. MSNA was suppressed to a similar extent (27%) at 10- and 20-mmHg LBPP. However, at 30-mmHg LBPP, MSNA tended to increase but was still nearly at the control value. Mean arterial pressure was elevated (11%), total peripheral resistance markedly increased (36%), and stroke volume and cardiac output tended to decrease at 30 mmHg LBPP. Heart rate remained unchanged throughout the procedures. Left atrial dimension significantly increased during 10- and 30-mmHg LBPP, indicating an increased cardiac filling. These results suggest that the inhibitory effect of the cardiopulmonary baroreflex on MSNA at 10- and 20-mmHg LBPP could be counteracted by the sympathoexcitatory effect of the intramuscular pressure sensitive mechanoreflex at 30-mmHg LBPP. However, the increment of total peripheral resistance at 30-mmHg LBPP may not depend exclusively on this small enhancement of MSNA. PMID- 9746474 TI - BDM drives protein dephosphorylation and inhibits adenine nucleotide exchange in cardiomyocytes. AB - Contractile dysfunction plays a key role in injury sustained by ischemic myocardium at reperfusion, whereas interventions that impede hypercontracture enhance recovery. In permeabilized adult rat cardiomyocytes, the negative inotrope 2,3-butanedione monoxime (BDM; 10-50 mM) inhibited rigor at low MgATP concentration but stimulated net ATP hydrolysis. Hydrolysis was attenuated by H 7, kaempferol, chelerythrine, and genistein. Evidently BDM opposed phosphorylation of both serine/threonine and tyrosine kinase target proteins, either directly or by enhancing protein phosphatase activity, in a futile cycle of ATP hydrolysis independent of cross-bridge cycling. Although 20 mM BDM did not affect the onset of rigor contracture in permeabilized cells at low MgATP, in intact cells exposed to the metabolic inhibitors cyanide and 2-deoxyglucose rigor onset was accelerated, indicating that BDM increases ATP depletion in quiescent cardiomyocytes. Conversely, in cells exposed to the mitochondrial uncoupler carbonyl cyanide p-trifluoromethoxyphenylhydrazone, BDM delayed the onset of contracture and hence ATP depletion, consistent with an inhibition of adenine nucleotide movement across the mitochondrial inner membrane. Such effects will limit the value of BDM as a cardioprotective agent at physiological temperature. PMID- 9746475 TI - Dissociation between regional dysfunction and beta-adrenergic receptor signaling in heart failure. AB - We have previously shown that left ventricular (LV) pacing-induced heart failure is associated with preserved wall thickening in the interventricular septum (IVS) compared with the posterolateral wall (PLW). The current study focuses on the relationship between regional myocardial function and altered beta-adrenergic receptor (beta-AR) signaling. We studied 15 pigs: 6 controls and 9 paced from the left ventricle (225 beats/min, 26 +/- 3 days). Heart failure was documented by decreased LV fractional shortening (P < 0.0001) and increased left atrial pressure (P < 0.0001). In heart failure, despite marked differences in basal regional function (percent wall thickening: IVS, 33 +/- 10% vs. PLW, 13 +/- 7%; P = 0.0003), there were no differences between the two regions in beta-AR responsiveness, measured by regional wall thickening in response to dobutamine infusion and any measurement of adrenergic signaling. Adenylyl cyclase activity, beta-AR number, and beta-AR/Gs coupling were markedly reduced in failing LV without regional differences. In animals with heart failure, LV G protein receptor kinase (GRK) isoform 2 content was unchanged and GRK5, the other major GRK isoform, was increased more than threefold (IVS, 0.51 +/- 0.20 vs. 0. 12 +/- 0.12 arbitrary densitometric units, P = 0.01; PLW, 0.47 +/- 0. 15 vs. 0.13 +/- 0.09 arbitrary densitometric units, P = 0.03), but again, there were no regional differences. These data indicate that systemic rather than regional factors govern LV adrenergic signaling and that regional adrenergic signaling abnormalities poorly predict wall thickening in the same regions. PMID- 9746476 TI - Effect of treadmill exercise on transmural distribution of blood flow in hypertrophied left ventricle. AB - Pressure-overload left ventricular (LV) hypertrophy (LVH) is associated with increased vulnerability to subendocardial hypoperfusion during exercise. Abnormal perfusion could be the result of failure of the coronary vessels to grow in proportion to the degree of myocyte hypertrophy or could be due to increased extravascular forces acting on the intramural coronary vasculature. This study assessed the contribution of extravascular forces by examining the effect of exercise on the distribution of myocardial blood flow when coronary vasomotor tone was abolished with a maximal vasodilating dose of intracoronary adenosine. One year after ascending aortic banding in six dogs, the LV-to-body weight ratio was 7.80 +/- 0.38 g/kg compared with 4.57 +/- 0.20 g/kg in nine normal dogs (P < 0.01). Under awake resting conditions blood flow in LVH hearts increased from 1.17 +/- 0.27 ml . min-1 . g-1 during basal conditions to 5.78 +/- 1.06 ml . min 1 . g-1 during adenosine (at a coronary pressure of 100 +/- 6 mmHg), whereas in normal dogs blood flow increased from 1.22 +/- 0.17 to 5.26 +/- 0.71 ml . min-1 . g-1 (at a coronary pressure of 62 +/- 4 mmHg). At rest the transmural distribution of blood flow during adenosine was not different between hypertrophied and normal hearts, with subendocardial-to-subepicardial (Endo-to Epi) blood flow ratios of 1. 01 +/- 0.09 and 1.14 +/- 0.13, respectively (P = not significant). During adenosine infusion, treadmill exercise to produce heart rates of 200-220 beats/min caused redistribution of blood flow away from the subendocardium that was much more marked in LVH (Endo-to-Epi blood flow ratio = 0.35 +/- 0.04) than in normal hearts (Endo-to-Epi blood flow ratio = 0.76 +/- 0.09, P < 0.05 vs. LVH). In comparison with normal, the exaggerated decrease in subendocardial blood flow produced by exercise in LVH hearts resulted from abnormally increased extravascular compressive forces, including a greater decrease in diastolic duration and an increase in LV end-diastolic pressure. PMID- 9746477 TI - Hydrogen peroxide relaxes porcine coronary arteries by stimulating BKCa channel activity. AB - It has been known for a number of years that neutrophils and macrophages secrete H2O2 while fighting disease, and the levels obtained within the vasculature under these conditions can reach several hundred micromolar. Because the effect of H2O2 on vascular smooth muscle is not fully understood, the present study examined the cellular effects of H2O2 on coronary arteries. Under normal ionic conditions, H2O2 relaxed arteries that were precontracted with prostaglandin F2alpha or histamine (EC50 = 252 +/- 22 microM). The effect of H2O2 was concentration dependent and endothelium independent. In contrast, H2O2 did not relax arteries contracted with 80 mM KCl, suggesting involvement of K+ channels. Single-channel patch-clamp recordings revealed that H2O2 increased the activity of the large conductance (119 pS), Ca2+- and voltage-activated K+ (BKCa) channel. This response was mimicked by arachidonic acid and inhibited by eicosatriynoic acid, a lipoxygenase blocker, suggesting involvement of leukotrienes. Further studies on intact arteries demonstrated that eicosatriynoic acid not only blocked the vasodilatory response to H2O2 but unmasked a vasoconstrictor effect that was reversed by blocking cyclooxygenase activity with indomethacin. These findings identify a novel effector molecule, the BKCa channel, which appears to mediate the vasodilatory effect of H2O2, and suggest that a single signaling pathway, arachidonic acid metabolism, can mediate the vasodilatory and vasoconstrictor effects of H2O2 and possibly other reactive oxygen species. PMID- 9746478 TI - Quantification of sympathetic and parasympathetic tones by nonlinear indexes in normotensive rats. AB - Because the use of spectral powers of blood pressure (BP) and R-R interval (RR) in the low (LF) and high frequencies (HF) to quantify sympathetic and parasympathetic activities is still under debate, we questioned whether nonlinear methods may give better results. The BP signal was recorded for 30 min before and after intravenous injection of hexamethonium (20 mg/kg), atropine (0.5 mg/kg), atenolol (1 mg/kg), and prazosin (1 mg/kg) in conscious, normotensive Wistar Kyoto rats. Three nonlinear indexes [percentage of recurrence, percentage of determinism, and length index (Lmax)] extracted from the recurrence plot method were used to analyze the BP signal. Sympathetic but not parasympathetic blockade reduced BP level and its LF component. RR increased and decreased after beta- and alpha-blockades, respectively. Hexamethonium increased HF, and atropine reduced LF, of RR. Sympathetic blockade and, in particular, alpha-sympathetic blockade increased nonlinear indexes of BP. In contrast, parasympathetic blockade by atropine increased nonlinear indexes of RR. These results suggest that, compared with spectral indexes, nonlinear indexes may be more specific markers of sympathetic and parasympathetic tones. PMID- 9746479 TI - Myocardial overexpression of GRK3 in transgenic mice: evidence for in vivo selectivity of GRKs. AB - Transgenic mice were generated with cardiac-specific overexpression of the G protein-coupled receptor kinase 3 (GRK3) to explore the in vivo role of this GRK in cardiac function. GRK3 is expressed in the heart along with the beta adrenergic receptor kinase (beta-ARK1) and GRK5. We have previously demonstrated that myocardial-targeted overexpression in transgenic mice of beta-ARK1 (Koch, W.J., H. A. Rockman, P. Samama, R. A. Hamilton, R. A. Bond, C. A. Milano, and R. J. Lefkowitz. Science 268: 1350-1353, 1995) or GRK5 (Rockman, H.A., D.-J. Choi, N. U. Rahman, S. A. Akhter, R. J. Lefkowitz, and W. J. Koch. Proc. Natl. Acad. Sci. USA 93: 9954-9959, 1996) results in significant attenuation of beta adrenergic signaling and in vivo cardiac function and selective desensitization of angiotensin (ANG) II-mediated cardiac responses. Surprisingly, myocardial overexpression of GRK3 resulted in normal biochemical signaling through beta adrenergic receptors (beta-ARs), and in vivo hemodynamic function in response to a beta-AR agonist was indistinguishable from that in nontransgenic controls. Furthermore, in vivo signaling and functional responses to ANG II were unaltered. However, myocardial thrombin signaling, as assessed by p42/p44 mitogen-activated protein (MAP) kinase activation, was significantly attenuated in GRK3 transgenic mouse hearts, indicating a distinct in vivo substrate specificity for GRK3. PMID- 9746480 TI - Functional evidence for alternative ANG II-forming pathways in hamster cardiovascular system. AB - Like human chymase, hamster chymase is an ANG II-forming enzyme, but pathophysiological roles of chymase are still unknown. We determined the functional conversion of ANG I and [Pro11, D-Ala12]ANG I, a chymase-selective substrate, to ANG II in the hamster cardiovascular system. ANG I and [Pro11, D Ala12]ANG I produced similar dose-dependent pressor responses in conscious hamsters. Captopril and CV-11974, an ANG II type 1 (AT1)-receptor antagonist, inhibited the responses to ANG I; in contrast, the pressor responses to [Pro11, D Ala12]ANG I were suppressed only by CV-11974. In the isolated aorta, captopril suppressed ANG I-induced contraction by 84%; administration of captopril with either chymostatin or aprotinin eliminated the contraction. [Pro11, D-Ala12]ANG I induced contraction was not affected by captopril but was attenuated by chymostatin (71%) and aprotinin (57%). CV-11974 abolished the responses to both substrates, whereas PD-123319, an AT2-receptor antagonist, had no effect. In homogenates of the aorta and heart, soybean trypsin inhibitor-inhibitable ANG II formation predominated over captopril- or aprotinin-inhibitable ANG II formation. These data suggest that [Pro11,D-Ala12]ANG I and part of ANG I were functionally converted to ANG II by chymase and other serine protease(s) in hamster vessels, inducing AT1-receptor-mediated vasoconstriction. Biochemical data supported a role for chymase in the alternative pathway. PMID- 9746482 TI - Rabbit polymorphonuclear leukocytes release a factor that causes constriction of the coronary vasculature. AB - Polymorphonuclear leukocytes (PMN) have been shown to have numerous vasoactive effects, particularly in large artery bioassays. This study shows that rabbit PMN passively release a contractile factor that constricts the coronary vasculature of isolated, Langendorff-perfused rabbit hearts. The mechanism of action of this factor does not involve inhibition of nitric oxide (NO), production of cyclooxygenase metabolites, 5-hydroxytryptamine, or endothelin, or the activation of alpha-adrenoceptors but is a Ca2+-dependent process, because the constriction is inhibited by the Ca2+-channel blocker amlodipine. The activity of this factor is significantly inhibited if it is pretreated with trypsin or heated to 90 degreesC for 10 min, and the active factor is concentrated in the retentate of 100-kDa cutoff centrifuge filters, indicating that the factor is a protein >100 kDa in size. This study shows that rabbit PMN spontaneously release a protein factor that causes constriction of isolated, perfused rabbit hearts by a NO independent but Ca2+-dependent mechanism. PMID- 9746481 TI - Effect of cross-linked hemoglobin transfusion on endothelial-dependent dilation in cat pial arterioles. AB - We determined whether addition of hemoglobin to the plasma would inhibit endothelial-dependent dilation in brain where tight endothelial junctions limit hemoglobin extravasation. Pial arteriolar diameter was measured by intravital microscopy through closed cranial windows in anesthetized cats either without transfusion (hematocrit = 32%) or after exchange transfusion with an albumin or sebacyl-cross-linked human hemoglobin solution (hematocrit = 18%). Dilation of small, medium, and large arterioles to acetylcholine and ADP was not significantly altered by hemoglobin transfusion. The dilatory responses were inhibited by the nitric oxide synthase inhibitor NG-nitro-L-arginine, although significant dilation to 30 microM acetylcholine persisted in small arterioles in the control and albumin-transfused group but not in the hemoglobin-transfused group. The dilatory response to the nitric oxide donor 3-morpholinosydnonimine was unaffected by albumin or hemoglobin transfusion, but the response to nitroprusside was reduced by one-third after hemoglobin transfusion. When cross linked hemoglobin was superfused through the cranial window, the acetylcholine response became inhibited at a hemoglobin concentration of 0.1 microM and was completely blocked at 10 microM. Because this concentration is substantially less than the 500 microM hemoglobin concentration in plasma after transfusion when there was no inhibition of the acetylcholine response, hemoglobin permeation of the blood-brain barrier was considered negligible. We conclude that exchange of red cell-based hemoglobin with plasma-based hemoglobin does not produce a more effective sink for endothelial-derived nitric oxide evoked by agonist receptor mediated activation. Furthermore, decreased hematocrit does not affect agonist evoked endothelial-dependent dilation. PMID- 9746483 TI - Ecto-5'-nucleotidase is not required for ischemic preconditioning in rabbit myocardium in situ. AB - This study tested the hypothesis that cardiac ecto-5'-nucleotidase (ecto-5'-NT) activity during ischemic preconditioning (PC) contributes to augmented tolerance against ischemia, thereby reducing infarct size in the rabbit heart in situ. The effects of alpha,beta-methylene-adenosine diphosphate (AOPCP), a selective inhibitor of ecto-5'-NT, on cardiovascular responses to AMP were measured to establish in vivo activities of the enzyme and its inhibitor. Left atrial infusion of AOPCP (0.75 mg . kg-1 . min-1) raised AOPCP plasma levels to 138 microM; under these conditions negative chronotropic and inotropic effects of AMP were blocked, demonstrating essentially full inhibition of ecto-5'-NT in the heart in situ. This AOPCP-blocked heart in situ model was used to examine the proposed contribution of ecto-5'-NT in ischemic PC. Myocardial infarction caused by 30-min ischemia was followed by 3-h reperfusion. Infarct size (IS) was measured and expressed as a percentage of the size of the area at risk (%IS/AR). In untreated controls, %IS/AR was 38.1 +/- 3.8%; PC (5-min ischemia, 5-min reperfusion) markedly reduced %IS/AR to 10.0 +/- 2.0%. Essentially identical IS reductions by PC were observed in AOPCP-blocked animals (%IS/AR = 13.8 +/- 2.2 and 13.3 +/- 1.8% in rabbits receiving AOPCP at 0.75 and 1.50 mg . kg-1 . min-1, respectively); here plasma AOPCP levels were established before and during PC but not during the subsequent prolonged ischemia. As expected, AOPCP also did not affect %IS/AR in non-PC controls (%IS/AR = 35.5 +/- 3.7%). In contrast but as predicted, adenosine-receptor blockade by 8-phenyltheophylline (10 mg/kg iv) substantially attenuated IS reduction by PC in both AOPCP-blocked and control hearts (%IS/AR = 25.2 +/- 4.3 and 21.8 +/- 2.2%, respectively; P < 0.05 vs. PC alone). The results demonstrate that cardiac ecto-5'-NT is not required for ischemic PC against infarction in the rabbit. PMID- 9746484 TI - Overexpression of alpha1B-adrenergic receptor induces left ventricular dysfunction in the absence of hypertrophy. AB - The stimulation of cardiac alpha1-adrenergic receptors (AR) modulates the heart's inotropic response and plays a role in the induction of cardiomyocyte hypertrophy. We have analyzed transgenic mouse lines overexpressing a wild-type alpha1B-AR specifically in the heart. Basal level systolic and diastolic left ventricular (LV) contractile function was depressed both in the anesthetized closed-chest mouse and the perfused working-heart preparation. Intrinsic LV function was further characterized under controlled preload and afterload conditions using the perfusion model. Contractile parameters were restored by chronic treatment with the alpha-AR antagonist prazosin. In ventricular function curves, the load-dependent force increases (length-tension effects) remained intact, although the transgenic curve was shifted to lower levels. The basal level contractile deficits were paralleled by a decrease in calcium transients in isolated LV cardiomyocytes. LV function comparable to controls was restored by isoproterenol stimulation. The physiological changes occurred in the absence of cardiomyocyte hypertrophy. This transgenic model will be useful for studying the potential role of alpha1-AR in cardiac contractility and hypertrophy. PMID- 9746485 TI - Intracellular acidosis differentially regulates KV channels in coronary and pulmonary vascular muscle. AB - Decreases in intracellular pH (pHi) potently dilate coronary resistance arteries but constrict small pulmonary arteries. To define the ionic mechanisms of these responses, this study investigated whether acute decreases in pHi differentially regulate K+ currents in single vascular smooth muscle (VSM) cells isolated from rat coronary and pulmonary resistance arteries. In patch-clamp studies, whole cell K+ currents were elicited by 10-mV depolarizing steps between -60 and 0 mV in VSM cells obtained from 50- to 150-micrometers-OD arterial branches, and pHi was lowered by altering the NH4Cl gradient across the cell membrane. Progressively lowering pHi from calculated values of 7.0 to 6.7 and 6.4 increased the peak amplitude of K+ current in coronary VSM cells by 15 +/- 5 and 23 +/- 3% but reduced K+ current in pulmonary VSM cells by 18 +/- 3 and 21 +/- 3%, respectively. These changes were reversed by returning cells to the control pHi of 7.0 and were eliminated by dialyzing cells with pipette solution containing 50 mmol/l HEPES to buffer NH4Cl-induced changes in pHi. Pharmacological block of ATP sensitive K+ channels and Ca2+-activated K+ channels by 1 micromol/l glibenclamide and 100 nmol/l iberiotoxin, respectively, did not prevent changes in K+ current levels induced by acidotic pHi. However, block of voltage-gated K+ channels by 3 mmol/l 4-aminopyridine abolished acidosis-induced changes in K+ current amplitudes in both VSM cell types. Interestingly, alpha-dendrotoxin (100 nmol/l), which blocks only select subtypes of voltage-gated K+ channels, abolished the acidosis-induced decrease in K+ current in pulmonary VSM cells but did not affect the acidosis-induced increase in K+ current observed in coronary VSM cells. These findings suggest that opposing, tissue-specific effects of pHi on distinct subtypes of voltage-gated K+ channels in coronary and pulmonary VSM membranes may differentially regulate vascular reactivity in these two circulations under conditions of acidotic stress. PMID- 9746486 TI - Effects of isoflurane on [Ca2+]i, SR Ca2+ content, and twitch force in intact trabeculae. AB - The goal was to test whether isoflurane exerts its depressant effect on the heart by mainly affecting the intracellular Ca2+ transient [Ca2+]i. Intact rat ventricular trabeculae, paced at 0.5 Hz and 30 degreesC with extracellular [Ca2+] ([Ca2+]o) of 2 mM, were used. The [Ca2+]i was monitored using fura 2 injected into the myoplasm. The sarcoplasmic reticulum (SR) Ca2+ content was estimated using rapid cooling with or without caffeine to induce Ca2+ release and contracture. A plot of peak twitch force versus peak [Ca2+]i transient with increasing isoflurane concentration declines linearly so that a 56% reduction in the peak [Ca2+]i transient would abolish twitch force. This relationship is intermediate between those obtained with lowering [Ca2+]o, which depresses twitch force through a reduction of the [Ca2+]i transient, and adding 2,3-butanedione monoxime, which reduces the responsiveness of the contractile system to [Ca2+]i. The isoflurane effect is different from that of halothane with respect to both the above relationship and the rapid-cooling response. Isoflurane abolishes the ability of rapid cooling to liberate Ca2+ from the SR. PMID- 9746487 TI - Angiotensin II-induced hypertrophy of adult rat cardiomyocytes is blocked by nitric oxide. AB - The aim of the present study was to test the hypothesis that bradykinin stimulated release of nitric oxide (NO) and/or prostacyclin from endothelium blocks myocyte hypertrophy in vitro. Angiotensin II increased [3H]phenylalanine incorporation by 21 +/- 2% in myocytes cocultured with endothelial cells; this was abolished by bradykinin in the presence of endothelial cells. Bradykinin increased cytosolic concentrations of cGMP by 29 +/- 4% in myocytes cocultured with endothelial cells. This was abolished by inhibition of NO synthase and by a cyclooxygenase inhibitor. Angiotensin II also increased [3H]phenylalanine incorporation by 28 +/- 3% in myocytes cultured in the absence of endothelial cells. This effect of angiotensin II in monoculture was abolished by donors of NO but not by bradykinin. Neither the stable analog of prostacyclin (iloprost) nor the prostacyclin second messanger analog 8-bromo-cAMP (8-BrcAMP) blocked the effect of angiotensin II. Furthermore, 8-BrcAMP and iloprost individually increased [3H]phenylalanine incorporation. The antihypertrophic effects of bradykinin are critically dependent on endothelium-derived NO. PMID- 9746489 TI - Nifedipine increases microvascular permeability via a direct local effect on postcapillary venules. AB - Calcium-channel antagonist drugs are prescribed widely for angina and hypertension. A limiting side effect is edema, which can make heart failure worse. We show that nifedipine, a dihydropyridine-type calcium-channel antagonist, can increase vascular permeability in rat skeletal muscle and skin when injected locally. In nifedipine-injected cremaster muscle, the copper content, used to quantify Monastral blue dye accumulation, was 15.0 +/- 2.4 microgram/g compared with 5.3 +/- 0.7 microgram/g in control preparations (P < 0. 05). The injection of nifedipine in rat skin in vivo increased local plasma leakage in injected sites from 5.5 +/- 1.1 microliter in control sites to 9.9 +/- 2.5, 17.0 +/- 2.4, 24.3 +/- 5.9, and 23.3 +/- 5.4 microliter in sites injected with 10(-10), 10(-9), 10(-8), or 10(-7.2) mol/site, respectively (P < 0.05 in each case compared with control). Vascular labeling techniques using light microscopy, electron microscopy, and microanalysis show that the microvascular site of leakage is not from capillaries but from postcapillary venules of 12-36 micrometer in diameter, the same site that controls the edema response in inflammation. Nifedipine can act within the microcirculation to increase the permeability of the postcapillary venule. PMID- 9746488 TI - Demonstration of an early and a late phase of ischemic preconditioning in mice. AB - It is unknown whether ischemic preconditioning (PC; either early or late) occurs in the mouse. The goal of this study was to answer this question and to develop a reliable and physiologically relevant murine model of both early and late ischemic PC. A total of 201 mice were used. In nonpreconditioned open-chest animals subjected to 30 min of coronary occlusion followed by 24 h of reperfusion, infarct size (tetrazolium staining) averaged 52% of the region at risk. When the 30-min occlusion was performed 10 min after a PC protocol consisting of six cycles of 4-min occlusion and 4-min reperfusion, infarct size was reduced by 75%, indicating an early PC effect. When the 30-min occlusion was performed 24 h after the same PC protocol, infarct size was reduced by 48%, indicating a late PC effect. In mice in which the 30-min occlusion was followed by 4 h of reperfusion, infarct size was similar to that observed after 24 h of reperfusion, indicating that a 4-h reperfusion interval is sufficient to detect the final extent of cell death in this model. Fundamental physiological variables (body temperature, arterial oxygenation, acid-base balance, heart rate, and arterial pressure) were measured and found to be within normal limits. Taken together, these results demonstrate that, in the mouse, a robust infarct-sparing effect occurs during both the early and the late phases of ischemic PC, although the early phase is more powerful. This murine model is physiologically relevant, provides reliable measurements, and should be useful for elucidating the cellular mechanisms of ischemic PC in genetically engineered animals. PMID- 9746490 TI - Localization of the ANG II type 2 receptor in the microcirculation of skeletal muscle. AB - Only functional studies have suggested the presence of the ANG II type 2 (AT2) receptor in the microcirculation. To determine the distribution of this receptor in the rat skeletal muscle microcirculation, a polyclonal rabbit anti-rat antiserum was developed and used for immunohistochemistry and Western blot analysis. The antiserum was prepared against a highly specific and antigenic AT2 receptor synthetic peptide and was validated by competition and sensitivity assays. Western blot analysis demonstrated a prominent, single band at approximately 40 kDa in cremaster and soleus muscle. Immunohistochemical analysis revealed a wide distribution of AT2 receptors throughout the skeletal muscle microcirculation in large and small microvessels. Microanatomic studies displayed an endothelial localization of the AT2 receptor, whereas dual labeling with smooth muscle alpha-actin also showed colocalization of the AT2 receptor with vascular smooth muscle cells. Other cells associated with the microvessels also stained positive for AT2 receptors. Briefly, this study confirms previous functional data and localizes the AT2 receptor to the microcirculation. These studies demonstrate that the AT2 receptor is present on a variety of vascular cell types and that it is situated in a fashion that would allow it to directly oppose ANG II type 1 receptor actions. PMID- 9746491 TI - Hemodynamics of gastric microcirculation in rats. AB - Recently, we described a novel preparation of rat stomach for vascular micropuncture studies. The aim of the present study was to directly measure basic microvascular parameters along the length of the gastric vasculature. Blood vessels were identified, and intravascular pressure was measured with a servo null transducer, vessel dimensions with videometry, blood flow with microspheres, and plasma colloid osmotic pressure with an osmometer. When systemic arterial pressure was 100-110 mmHg, intravascular pressures in small arteries, primary, secondary, and tertiary submucosal arterioles, mucosal terminal arterioles, and muscle arterioles were 77.8 +/- 2.6, 74.6 +/- 2.5, 54.1 +/- 1.8, 34.4 +/- 1.6, 32.4 +/- 1.2, and 30.5 +/- 1.4 (SE) mmHg, respectively. Intravascular pressures in collecting veins, secondary and primary submucosal venules, muscle venules, and small veins were 26.6 +/- 1.1, 21.8 +/- 1.6, 17.1 +/- 0. 8, 18.2 +/- 0.9, and 14.4 +/- 0.6 mmHg, respectively. Capillary pressure in the mucosa (28 mmHg), as estimated by interpolation between terminal arteriole and collecting venule pressures, was significantly higher than in the muscle layer (23.6 +/- 1.4 mmHg). A total of 155 vessels from 25 animals were sampled. Relative blood flows were 16 +/- 3% in the muscle and 84 +/- 3% in the mucosa-submucosa. Analysis of filtration forces in these two different capillary beds suggests that gastric mucosal capillaries are primarily a filtering network, whereas muscle capillaries are in fluid balance. Calculated resistance ratios indicate low precapillary but relatively high postcapillary vascular resistance in the gastric mucosa. PMID- 9746493 TI - Mutual information discloses relationship between hemodynamic variables in artificial heart-implanted dogs. AB - A mutual information (MI) method for assessment of the relationship between hemodynamic variables was proposed and applied to the analysis of heart rate (HR), arterial blood pressure (BP), and renal sympathetic nerve activity (RSNA) in artificial heart-implanted dogs to quantify correlation between these parameters. MI measures the nonlinear as well as linear dependence of two variables. Simulation studies revealed that this MI technique furnishes mathematical features well suited to the investigation of nonlinear dynamics such as the cardiovascular system and can quantify a relationship between two parameters. To constitute a model free of the natural heart, two pneumatically actuated ventricular assist devices were implanted as biventricular bypasses in acute canine experiments. RSNA was detected with the use of bipolar electrodes attached to the renal sympathetic nerve. Analysis of data during control revealed that correlation between HR and RSNA was higher than that between HR and BP and that between RSNA and BP (P < 0.05). Although RSNA seemed to fluctuate noncorrelatedly with BP in higher pacing rates, the MI values between them disclosed their strong correlation. Surprisingly, correlation between RSNA and BP was stronger during a pacing rate of 60 beats/min than during higher pacing rates and control (P < 0. 05). It is suggested that the baroreflex system may be susceptible to pacing rates during the total artificial heart state. We calculated the time delay between HR and RSNA, between RSNA and BP, and between HR and BP by regarding a time delay at which the maximum MI value between each pair of parameters was given as a physiological delay. Our results indicate that RSNA leads BP, BP leads HR, and RSNA leads HR during control (P < 0.05). We conclude that this method could provide a powerful means for measuring correlation of physiological variables. PMID- 9746492 TI - Effects of coronary artery disease on expression and microvascular response to VEGF. AB - The effects of coronary artery disease (CAD) on human coronary microvascular responses to vascular endothelial growth factor (VEGF) and the alterations of the myocardial expressions of VEGF and its flk-1 and flt-1 receptors were examined in 48 patients. Microvascular studies were performed in vitro with video microscopy. The expressions of VEGF and its receptors were examined using Northern analysis of total mRNA, and the expressions of constitutive nitric oxide synthase (cNOS) and inducible nitric oxide synthase (iNOS) were examined by RT-PCR. VEGF and hepatocyte growth factor (HGF) caused potent relaxations of microvessels. These responses were reduced in the presence of NG-nitro-L-arginine and the tyrosine kinase inhibitor genistein or in microvessels from patients with CAD. Relaxations to substance P and sodium nitroprusside were similar in both groups. The substance P response was abolished in the presence of NG-nitro-L-arginine. The expression of VEGF and its receptors and the expression of cNOS and iNOS were not altered in patients with CAD. In conclusion, VEGF and HGF elicit the release of nitric oxide through activation of tyrosine kinase receptors. CAD is associated with reduced vascular responses to both VEGF and HGF; this is not likely due to a reduced expression of VEGF or flt-1 or flk-1 receptors and not due to a generalized endothelium dysfunction despite the presence of mild hypercholesterolemia in these patients with CAD. These findings may have important implications regarding the efficacy of endogenous and exogenous VEGF in patients with risk factor for CAD. PMID- 9746494 TI - Sensitivity of canine intrinsic cardiac neurons to H2O2 and hydroxyl radical. AB - To determine whether intrinsic cardiac neurons are sensitive to oxygen-derived free radicals in situ, studies were performed in 44 open-chest anesthetized dogs. 1) When H2O2 (600 microM) was administered to right atrial neurons of 36 dogs via their local arterial blood supply, neuronal activity either increased (+92% in 16 dogs) or decreased (-61% in 20 dogs), depending on the population of neurons studied. H2O2 (600 microM) administered into the systemic circulation did not affect neuronal activity, measured cardiac indexes, or aortic pressure. 2) The iron-chelating agent deferoxamine (20 mg/kg iv), a chemical that prevents the formation of oxygen-derived free radicals, reduced the activity generated by neurons (-57%) in 8 of 10 dogs. 3) H2O2 did not affect neuronal activity when administered in the presence of deferoxamine in these 10 dogs. 4) When the ATP sensitive potassium (KATP) channel opener cromakalim (20 microM) was administered to intrinsic cardiac neurons in another 21 animals via their regional arterial blood supply, ongoing neuronal activity in 15 of these dogs decreased by 54%. 5) Neuronal activity was not affected by H2O2 when administered in the presence of cromakalim in 16 dogs. These data indicate that 1) some intrinsic cardiac neurons are sensitive to exogenous H2O2, 2) such neurons are tonically influenced by locally produced oxygen-derived free radicals in situ, and 3) intrinsic cardiac neurons possess KATP channels that are functionally important during oxidative challenge. PMID- 9746495 TI - Abnormal myocyte Ca2+ homeostasis in rabbits with pacing-induced heart failure. AB - To determine whether there are abnormalities in myocyte excitation-contraction coupling and intracellular Ca2+ concentration ([Ca2+]i) homeostasis in pacing induced heart failure (PF), we measured L-type Ca2+ current (ICa,L) and Na+/Ca2+ exchanger current (INa/Ca) with voltage clamp and measured intracellular Na+ concentration ([Na+]i) and [Ca2+]i with the use of sodium-binding benzofuran isophthalate (SBFI) and fluo 3 in ventricular myocytes isolated from control and paced rabbits. The peak systolic and diastolic levels and the amplitude of electrically stimulated [Ca2+]i transients (0.25 Hz, extracellular Ca2+ concentration = 1.08 mM) were significantly less in PF myocytes. Also, there was prolongation of the times to peak and decline of [Ca2+]i transients. ICa,L density was markedly decreased in PF myocytes. INa/Ca at -40 mV elicited by rapid exposure to 0 Na+ solution with a rapid solution switcher was significantly reduced in PF myocytes, suggesting that the function of the Na+/Ca2+ exchanger is impaired in these myocytes. In PF myocytes the decline of the [Ca2+]i transient when the Na+/Ca2+ exchanger was abruptly disabled was markedly prolonged compared with the decline in control myocytes, consistent with depressed sarcoplasmic reticulum (SR) Ca2+-ATPase function. RNase protection assay showed decreased levels of Na+/Ca2+ exchanger and SR Ca2+-ATPase mRNA in PF hearts, consistent with the function studies. We conclude that the functions of L-type Ca2+ channels, Na+/Ca2+ exchanger, and SR Ca2+-ATPase are impaired in myocytes from rabbit hearts with failure induced by rapid pacing. These abnormalities result in reduced [Ca2+]i transients and systolic and diastolic dysfunction and appear to account for the abnormal ventricular function observed. PMID- 9746496 TI - Role of K+ channels in arteriolar vasodilation mediated by integrin interaction with RGD-containing peptide. AB - Integrins are transmembrane adhesion receptors found on most cells, including vascular smooth muscle cells. Several integrins bind to the conserved amino acid sequence Arg-Gly-Asp (RGD), and synthetic RGD-containing peptides can cause endothelium-independent arteriolar vasodilation by interacting with the alphavbeta3-integrin expressed by vascular smooth muscle. We hypothesized that RGD peptide-induced vasodilation involves K+ channels. Rat cremaster arterioles were treated with cRGD (GPenGRGDSPCA) in the presence or absence of the nonselective K+ channel inhibitor tetraethylammonium (TEA, 20 mM). TEA caused arterioles to constrict by 19 +/- 5% and inhibited cRGD-induced vasodilation (n = 7, P < 0.05). Vessels preconstricted with phenylephrine (5 x 10(-7) M) showed no significant inhibition of the dilatory response to cRGD, indicating that inhibition by TEA was not related to increased vasomotor tone. Further evidence for the involvement of K+ channels was obtained by addition of 100 mM KCl (n = 5), which inhibited vasodilation caused by cRGD. Inhibition of large and small conductance, Ca2+-activated K+ channels with iberiotoxin (100 nM) or apamin (25 nM), respectively, had no effect on cRGD-induced vasodilation. Partial inhibition of vasodilation was observed with inhibitors of voltage-gated (4-aminopyridine, 1 mM), ATP-sensitive (glibenclamide, 1 microM), and inward rectifying (barium, 50 microM) K+ channels. These data support the hypothesis that integrin-signaling pathways leading to arteriolar vasodilation may involve modulation of K+ channel function. PMID- 9746497 TI - A nonlinear explanation of aging-induced changes in heartbeat dynamics. AB - The possibility of computing a cardiac age on the basis of spectral analysis of healthy individual tachograms was confirmed and facilitated by the use of a nonlinear technique: recurrence quantification analysis. The age of 112 subjects was predicted by this technique in terms of a progressive increase in the deterministic character of the heartbeat. This result confirms the "random-walk" character of the heartbeat as predicted by the terminal dynamics paradigm, thus allowing for a simple and comprehensive model of the effect of aging on cardiac dynamics: as age progresses, heart rate dynamics become increasingly predictable (constrained) on a beat-to-beat basis. This implies a basically stochastic nature of heart rate dynamics, probably reflecting the continuous adjustments to an unpredictable internal environment. PMID- 9746498 TI - Stimulation of different phospholipase A2 isoforms by TNF-alpha and IL-1beta in adult rat ventricular myocytes. AB - We previously showed that in adult rat ventricular myocytes interleukin (IL) 1beta activates a membrane-associated, Ca2+-independent phospholipase A2 (iPLA2). In this study, we examined the possible existence of different PLA2 isoforms and effects of tumor necrosis factor (TNF)-alpha on iPLA2 activities. Western blot analysis identified iPLA2 in both membrane (approximately 82 kDa) and cytosolic (approximately 40 kDa) fractions and identified Ca2+-dependent PLA2 (cPLA2) only in cytosolic fractions. With plasmenylcholine or alkylacyl glycerophosphorylcholine as substrate, TNF-alpha elicited a twofold transient increase in cytosolic iPLA2 activity accompanied by an increase in arachidonic acid release and decreased membrane-associated iPLA2 activity with plasmenylcholine. With phosphatidylcholine as substrate, TNF-alpha decreased both cytosolic and membrane-associated iPLA2 activities. TNF-alpha-induced increases in cytosolic iPLA2 activity and arachidonic acid release were completely blocked by methyl arachidonyl fluorophosphonate (MAFP) but not by bromoenol lactone (BEL). TNF-alpha and IL-1beta together enhanced synergistically cytosolic and membrane PLA2 activities and arachidonic acid release that were blocked differentially by MAFP and BEL, respectively, and inhibited completely by MAFP plus BEL. These results suggest that TNF-alpha and IL-1beta act on different PLA2 isoforms in ventricular myocytes. PMID- 9746499 TI - Role of intracellular Ca2+ and pH in positive inotropic response of cardiomyocytes to diacylglycerol. AB - Diacylglycerol has been hypothesized to mediate the positive inotropic response of myocardium to the alpha-adrenergic agonists angiotensin II and endothelin. The mechanism of action of diacylglycerol was examined here in adult rat ventricular myocytes by releasing dioctanoylglycerol (diC8) intracellularly from a caged compound while monitoring Ca2+ transients and pH with fluorescent indicators. DiC8 caused a three- to fourfold increase in twitch amplitude and a twofold increase in the systolic Ca2+ transient. No other parameter was consistently influenced by diC8, including the kinetics of Ca2+ cycling, the Ca2+ content of the sarcoplasmic reticulum, or the myofilament Ca2+ sensitivity. DiC8 also had no detectable effect on intracellular pH or Na+/H+ antiport activity. Consistent with this finding, the Na+/H+ exchange inhibitor N-ethylisopropyl amiloride was without effect on the positive inotropic response to diC8. The marked enhancement of systolic Ca2+ by diC8 suggests that the process of excitation-contraction coupling is an important and possibly preferred target of diacylglycerol-protein kinase C signaling in myocardium. PMID- 9746500 TI - Endothelial dysfunction in human intramyocardial small arteries in atherosclerosis and hypercholesterolemia. AB - Vascular responses of human intramyocardial small arteries were examined in vitro to assess the influence of atherosclerosis and risk factors for coronary artery disease on endothelium-dependent relaxation. Recipient hearts were obtained from patients with ischemic (n = 14) and nonischemic (n = 13) cardiomyopathy undergoing heart transplantation. Small intramyocardial coronary arteries (mean internal diameter 313 +/- 11 micrometers) were mounted on a wire myograph for measurement of morphology and isometric tension. Vasodilation was examined after preconstriction with U-46619, a thromboxane A2 analog. Endothelium-dependent relaxation to acetylcholine and bradykinin was impaired in patients with ischemic compared with nonischemic cardiomyopathy (P < 0.01 and P < 0.001, respectively). Endothelium-independent relaxation to sodium nitroprusside was preserved. Incubation with L-arginine (3 mmol/l) did not improve endothelium-dependent relaxation to acetylcholine or bradykinin. With the use of stepwise multivariate analysis, hypercholesterolemia, but no other risk factor for atherosclerosis, was independently associated with impaired endothelium-dependent relaxation to acetylcholine (r = -0.50, P = 0.05) but not to bradykinin. Endothelial dysfunction in intramyocardial small arteries may predispose patients with nonobstructive epicardial atherosclerosis and hypercholesterolemia to myocardial ischemia. PMID- 9746501 TI - Capillaries and arterioles are electrically coupled in hamster cheek pouch. AB - In this report we demonstrate electrical communication in the microcirculation between arterioles and capillary networks in situ. Microvessel networks in the hamster cheek pouch, which included capillaries and their feeding arterioles, were labeled with the voltage-sensitive dye di-8-ANEPPS by intraluminal perfusion through a micropipette. Pulses of 140 mM potassium solution were applied by pressure ejection from micropipettes positioned on arterioles several hundred micrometers upstream from capillaries. Potassium caused membrane potential changes of 3-11 mV in capillary segments up to 1,200 micrometers distal to the stimulation site, with time delays of <1 s. Capillary membrane potential changes were biphasic, with initial depolarizations followed by hyperpolarizations. The size of the response decreased exponentially with the distance between the arteriole and capillary, with a 1/e distance of 590 micrometers. The time to peak depolarization of both arteriolar and capillary segments was similar. The time to peak response was significantly faster than that for responses from direct stimulation of capillaries. Capillary responses were also obtained when blood flow was either blocked or directed toward sites of stimulation. Acetylcholine (10(-4) M) and phenylephrine (10(-5) M) applied to the arterioles by iontophoresis produced monophasic hyperpolarizing and depolarizing responses, respectively, in capillaries with <1-s delay between stimulus and onset of the membrane potential change. These results provide evidence in situ of a pathway for electrical communication between arteriolar and capillary levels of the microcirculation. PMID- 9746502 TI - Mechanosensitive ion channels in putative aortic baroreceptor neurons. AB - Cell-attached patch-clamp experiments were performed on dissociated neurons from nodose ganglia of adult rats. Putative aortic baroreceptor neurons were identified by labeling nerve endings in the adventitia of the aortic arch with the carbocyanine dye DiI. Whereas previous experiments demonstrated the presence of mechanosensitive (MS) whole cell currents, these experiments studied single MS ion channels and examined the influence of culture conditions on their expression. Single MS channels were activated by applying negative pressure through the recording pipette. Channel openings became more frequent as the negative pressure was increased, with open probability increasing significantly above 30 mmHg. MS channels had a slope conductance of 114 pS and a reversal potential of approximately 0 mV, consistent with a nonspecific cation conductance. Channels were not affected by antagonists of voltage-gated conductances but were blocked by 20 microM gadolinium, a known blocker of MS ion channels. When nodose neurons were cocultured with aortic endothelial cells, but not aortic smooth muscle cells, the percentage of patches exhibiting MS ion channels increased significantly, suggesting that aortic endothelial cells secrete a diffusible factor that increases channel expression. PMID- 9746503 TI - A tribute to Fredric Stewart Fay: June 5, 1943 - March 18, 1997. PMID- 9746504 TI - Molecular dynamics simulation of melittin in a dimyristoylphosphatidylcholine bilayer membrane. AB - Molecular dynamics trajectories of melittin in an explicit dimyristoyl phosphatidylcholine (DMPC) bilayer are generated to study the details of lipid protein interactions at the microscopic level. Melittin, a small amphipathic peptide found in bee venom, is known to have a pronounced effect on the lysis of membranes. The peptide is initially set parallel to the membrane-solution interfacial region in an alpha-helical conformation with unprotonated N-terminus. Solid-state nuclear magnetic resonance (NMR) and polarized attenuated total internal reflectance Fourier transform infrared (PATIR-FTIR) properties of melittin are calculated from the trajectory to characterize the orientation of the peptide relative to the bilayer. The residue Lys7 located in the hydrophobic moiety of the helix and residues Lys23, Arg24, Gln25, and Gln26 at the C-terminus hydrophilic form hydrogen bonds with water molecules and with the ester carbonyl groups of the lipids, suggesting their important contribution to the stability of the helix in the bilayer. Lipid acyl chains are closely packed around melittin, contributing to the stable association with the membrane. Calculated density profiles and order parameters of the lipid acyl chains averaged over the molecular dynamics trajectory indicate that melittin has effects on both layers of the membrane. The presence of melittin in the upper layer causes a local thinning of the bilayer that favors the penetration of water through the lower layer. The energetic factors involved in the association of melittin at the membrane surface are characterized using an implicit mean-field model in which the membrane and the surrounding solvent are represented as structureless continuum dielectric material. The results obtained by solving the Poisson Bolztmann equation numerically are in qualitative agreement with the detailed dynamics. The influence of the protonation state of the N-terminus of melittin is examined. After 600 ps, the N-terminus of melittin is protonated and the trajectory is continued for 400 ps, which leads to an important penetration of water molecules into the bilayer. These observations provide insights into how melittin interacts with membranes and the mechanism by which it enhances their lysis. PMID- 9746505 TI - Photochemistry in dried polymer films incorporating the deionized blue membrane form of bacteriorhodopsin. AB - The preparation and photochemical properties of dried deionized blue membrane (dIbR600; lambdamax approximately 600 nm, epsilon approximately 54, 760 cm-1 M-1, f approximately 1.1) in polyvinyl alcohol films are studied. Reversible photoconversion from dIbR600 to the pink membrane (dIbR485; lambdamax approximately 485 nm) is shown to occur in these films under conditions of strong 647-nm laser irradiation. The pink membrane analog, dIbR485, has a molar extinction coefficient of approximately 39,000 cm-1 M-1 (f approximately 1.2). The ratio of pink --> blue and blue --> pink quantum efficiencies is 33 +/- 5. We observe an additional blue-shifted species (dIbR455, lambdamax approximately 455 nm) with a very low oscillator strength (f approximately 0.6, epsilon approximately 26,000 cm-1 M-1). This species is the product of fast thermal decay of dIbR485. Molecular modeling indicates that charge/charge and charge/dipole interactions introduced by the protonation of ASP85 are responsible for lowering the excited-state all-trans --> 9-cis barrier to approximately 6 kcal mol-1 while increasing the corresponding all-trans --> 13-cis barrier to approximately 4 kcal mol-1. Photochemical formation of both 9-cis and 13-cis photoproducts are now competitive, as is observed experimentally. We suggest that dIbR455 may be a 9 cis, 10-s-distorted species that partially divides the chromophore into two localized conjugated segments with a concomitant blue shift and decreased oscillator strength of the lambdamax absorption band. PMID- 9746506 TI - Two-dimensional determination of the cellular Ca2+ binding in bovine chromaffin cells. AB - The spatiotemporal profile of intracellular calcium signals is determined by the flux of calcium ions across different biological membranes as well as by the diffusional mobility of calcium and different calcium buffers in the cell. To arrive at a quantitative understanding of the determinants of these signals, one needs to dissociate the flux contribution from the redistribution and buffering of calcium. Since the cytosol can be heterogeneous with respect to its calcium buffering property, it is essential to assess this property in a spatially resolved manner. In this paper we report on two different methods to estimate the cellular calcium binding of bovine adrenal chromaffin cells. In the first method, we use voltage-dependent calcium channels as a source to generate calcium gradients in the cytosol. Using imaging techniques, we monitor the dissipation of these gradients to estimate local apparent calcium diffusion coefficients and, from these, local calcium binding ratios. This approach requires a very high signal-to-noise ratio of the calcium measurement and can be used when well defined calcium gradients can be generated throughout the cell. In the second method, we overcome these problems by using calcium-loaded DM-nitrophen as a light-dependent calcium source to homogeneously and quantitatively release calcium in the cytosol. By measuring [Ca2+] directly before and after the photorelease process and knowing the total amount of calcium being released photolytically, we get an estimate of the fraction of calcium ions which does not appear as free calcium and hence must be bound to either the indicator dye or the endogenous calcium buffer. This finally results in a two-dimensional map of the distribution of the immobile endogenous calcium buffer. We did not observe significant variations of the cellular calcium binding at a spatial resolution of approximately 2 micron. Furthermore, the calcium binding is not reduced by increasing the resting [Ca2+] to levels as high as 1.1 microM. This is indicative of a low calcium affinity of the corresponding buffers and is in agreement with a recent report on the affinity of these buffers (Xu, T., M. Naraghi, H. Kang, and E. Neher. 1997. Biophys. J. 73:532-545). In contrast to the homogeneous distribution of the calcium buffers, the apparant calcium diffusion coefficient did show inhomogeneities, which can be attributed to restricted diffusion at the nuclear envelope and to rim effects at the cell membrane. PMID- 9746507 TI - Intracellular fluorescent probe concentrations by confocal microscopy. AB - A general method is described that takes advantage of the optical sectioning properties of a confocal microscope to enable measurement of both absolute and relative concentrations of fluorescent molecules inside cells. For compartments within cells that are substantially larger than the point spread function, the fluorescence intensity is simply proportional to the concentration of the fluorophore. For small compartments, the fluorescence intensity is diluted by contributions from regions outside the compartment. Corrections for this dilution can be estimated via calibrations that are based on the intensity distribution found in a computationally synthesized model for a cell or organelle that has been blurred by convolution with the microscope point spread function. The method is illustrated with four test cases: estimation of intracellular concentration of a fluorescent calcium indicator; estimation of the relative distribution between the neurite and soma of a neuronal cell of the InsP3 receptor on the endoplasmic reticulum; estimation of the distribution of the bradykinin receptor along the surface of a neuronal cell; and relative distribution of a potentiometric dye between the mitochondria and cytosol as a means of assaying mitochondrial membrane potential. PMID- 9746508 TI - 4Pi-confocal imaging in fixed biological specimens. AB - By combining the wavefronts produced by two high-aperture lenses, two-photon 4Pi confocal microscopy allows three-dimensional imaging of transparent biological specimens with axial resolution in the 100-140-nm range. We reveal the imaging properties of a two-photon 4Pi-confocal microscope as applied to a fixed cell. We demonstrate that a fast, linear point deconvolution suffices to achieve axially superresolved 3D images in the cytoskeleton. Furthermore, we describe stringent algorithms for alignment and control of the two lenses. We also show how to compensate for the effects of a potential refractive index mismatch of the mounting medium with respect to the immersion system. PMID- 9746509 TI - Calcium waves induced by large voltage pulses in fish keratocytes. AB - Intracellular calcium waves in fish keratocytes are induced by the application of electric field pulses with amplitudes between 55 and 120 V/cm and full width at half-maximum of 65-100 ms. Calcium concentrations were imaged using two-photon excited fluorescence microscopy (Denk et al., 1990 Science. 248:73-76; Williams et al. 1994 FASEB J. 8:804-813) and the ratiometric calcium indicator indo-1. The applied electric field pulses induced waves with fast calcium rise times and slow decays, which nucleated in the lamellipodium at the hyperpolarized side of the cells and, less frequently, at the depolarized side. The effectiveness of wave generation was determined by the change induced in the membrane potential, which is about half the field strength times the cell width in the direction of the field. Stimulation of waves began at voltage drops across the cell above 150 mV and saturated at voltage drops above 300 mV, where almost all cells exhibited a wave. Waves were not induced in low-calcium media and were blocked by the nonselective calcium channel blockers cobalt chloride and verapamil, but not by specific organic antagonists of voltage-sensitive calcium channel conductance. Thapsigargin stopped wave propagation in the cell body, indicating that calcium release from intracellular stores is necessary. Thus a voltage pulse stimulates Ca2+ influx through calcium channels in the plasma membrane, and if the intracellular calcium concentration reaches a threshold, release from intracellular stores is induced, creating a propagating wave. These observations and the measured parameters (average velocity approximately 66 micron/s and average rise time approximately 68 ms) are consistent with a wave amplification model in which[equation, see text] determines the effective diffusivity of the propagating molecules, D approximately 300 micron2/s (Meyer, 1991. Cell. 64:675 678). PMID- 9746510 TI - Analysis of synaptic transmission in the neuromuscular junction using a continuum finite element model. AB - There is a steadily growing body of experimental data describing the diffusion of acetylcholine in the neuromuscular junction and the subsequent miniature endplate currents produced at the postsynaptic membrane. To gain further insights into the structural features governing synaptic transmission, we have performed calculations using a simplified finite element model of the neuromuscular junction. The diffusing acetylcholine molecules are modeled as a continuum, whose spatial and temporal distribution is governed by the force-free diffusion equation. The finite element method was adopted because of its flexibility in modeling irregular geometries and complex boundary conditions. The resulting simulations are shown to be in accord with experiment and other simulations. PMID- 9746511 TI - Three electronic state model of the primary phototransformation of bacteriorhodopsin. AB - The primary all-trans --> 13-cis photoisomerization of retinal in bacteriorhodopsin has been investigated by means of quantum chemical and combined classical/quantum mechanical simulations employing the density matrix evolution method. Ab initio calculations on an analog of a protonated Schiff base of retinal in vacuo reveal two excited states S1 and S2, the potential surfaces of which intersect along the reaction coordinate through an avoided crossing, and then exhibit a second, weakly avoided, crossing or a conical intersection with the ground state surface. The dynamics governed by the three potential surfaces, scaled to match the in situ level spacings and represented through analytical functions, are described by a combined classical/quantum mechanical simulation. For a choice of nonadiabatic coupling constants close to the quantum chemistry calculation results, the simulations reproduce the observed photoisomerization quantum yield and predict the time needed to pass the avoided crossing region between S1 and S2 states at tau1 = 330 fs and the S1 --> ground state crossing at tau2 = 460 fs after light absorption. The first crossing follows after a 30 degrees torsion on a flat S1 surface, and the second crossing follows after a rapid torsion by a further 60 degrees. tau1 matches the observed fluorescence lifetime of S1. Adjusting the three energy levels to the spectral shift of D85N and D212N mutants of bacteriorhodospin changes the crossing region of S1 and S2 and leads to an increase in tau1 by factors 17 and 10, respectively, in qualitative agreement with the observed increase in fluorescent lifetimes. PMID- 9746512 TI - Effect of syncytium structure of receptor systems on stochastic resonance induced by chaotic potential fluctuation. AB - To study a role of syncytium structure of sensory receptor systems in the detection of weak signals through stochastic resonance, we present a model of a receptor system with syncytium structure in which receptor cells are interconnected by gap junctions. The apical membrane of each cell includes two kinds of ion channels whose gating processes are described by the deterministic model. The membrane potential of each cell fluctuates chaotically or periodically, depending on the dynamical state of collective channel gating. The chaotic fluctuation of membrane potential acts as internal noise for the stochastic resonance. The detection ability of the system increases as the electric conductance between adjacent cells generated by the gap junction increases. This effect of gap junctions arises mainly from the fact that the synchronization of chaotic fluctuation of membrane potential between the receptor cells is strengthened as the density of gap junctions is increased. PMID- 9746513 TI - Electroreceptor model of the weakly electric fish Gnathonemus petersii. I. The model and the origin of differences between A- and B-receptors. AB - We present an electroreceptor model of the A- and B-receptors of the weakly electric fish Gnathonemus petersii. The model consists of a sensory cell, whose membrane is separated into an apical and basal portions by support cells, and an afferent fiber. The apical membrane of the cell contains only leak channels, while the basal membrane contains voltage-sensitive Ca2+ channels, voltage sensitive and Ca2+-activated K+ channels, and leak channels. The afferent fiber is described with the modified Hodgkin-Huxley equation, in which the voltage sensitive gate of the K+ channels is a dynamic variable. In our model we suggest that the electroreceptors detect and process the information provided by an electric organ discharge (EOD) as follows: the current caused by an EOD stimulus depolarizes the basal membrane to a greatly depolarized state. Then the release of transmitter excites the afferent fiber to oscillate after a certain time interval. Due to the resistance-capacitance structure of the cells, they not only perceive the EOD intensity, but also sense the variation of the EOD waveform, which can be strongly distorted by the capacitive component of an object. Because of the different morphologies of A- and B-cells, as well as the different conductance of leak ion channels in the apical membrane and the different capacitance of A- and B-cells, A-receptors mainly respond to the EOD intensity, while B-receptors are sensitive to the variation of EOD waveform. PMID- 9746514 TI - Mutations in the EF-hand motif impair the inactivation of barium currents of the cardiac alpha1C channel. AB - Calcium-dependent inactivation has been described as a negative feedback mechanism for regulating voltage-dependent calcium influx in cardiac cells. Most recent evidence points to the C-terminus of the alpha1C subunit, with its EF-hand binding motif, as being critical in this process. The EF-hand binding motif is mostly conserved between the C-termini of six of the seven alpha1 subunit Ca2+ channel genes. The role of E1537 in the C-terminus of the alpha1C calcium channel inactivation was investigated here after expression in Xenopus laevis oocytes. Whole-cell currents were measured in the presence of 10 mM Ba2+ or 10 mM Ca2+ after intracellular injection of 1,2-bis(2-aminophenoxy)ethane-N,N,N',N' tetraacetic acid. Against all expectations, our results showed a significant reduction in the rate of voltage-dependent inactivation as measured in Ba2+ solutions for all E1537 mutants, whereas calcium-dependent inactivation appeared unscathed. Replacing the negatively charged glutamate residue by neutral glutamine, glycine, serine, or alanine significantly reduced the rate of Ba2+ dependent inactivation by 1.5-fold (glutamine) to 3.5-fold (alanine). The overall rate of macroscopic inactivation measured in Ca2+ solutions was also reduced, although a careful examination of the distribution of the fast and slow time constants suggests that only the slow time constant was significantly reduced in the mutant channels. The fast time constant, the hallmark of Ca2+-dependent inactivation, remained remarkably constant among wild-type and mutant channels. Moreover, inactivation of E1537A channels, in both Ca2+ and Ba2+ solutions, appeared to decrease with membrane depolarization, whereas inactivation of wild type channels became faster with positive voltages. All together, our results showed that E1537 mutations impaired voltage-dependent inactivation and suggest that the proximal part of the C-terminus may play a role in voltage-dependent inactivation in L-type alpha1C channels. PMID- 9746516 TI - Evidence for dimer participation and evidence against channel mechanism in A23187 mediated monovalent metal ion transport across phospholipid vesicular membrane. AB - The decay of the pH difference (DeltapH) across soybean phospholipid vesicular membrane by ionophore A23187 (CAL)-mediated H+/M+ exchange (M+ = Li+, Na+, K+, and Cs+) has been studied in the pH range 6-7.6. The DeltapH in these experiments were created by temperature jump. The observed dependence of DeltapH relaxation rate 1/tau on the concentration of CAL, pH, and the choice of M+ in vesicle solutions lead to the following conclusions. 1) The concentrations of dimers and other oligomers of A23187 in the membrane are small compared to the total concentration of A23187 in the membrane, similar to that in chloroform solutions reported in the literature. 2) In the H+ transport cycle leading to DeltapH decay, the A23187-mediated H+ translocation across the membrane is a fast step, and the rate-limiting step is the A23187-mediated M+ translocation. 3) Even though the monomeric Cal-H is the dominant species translocating H+, Cal-M is not the dominant species translocating M+ (even at concentrations higher than [Cal H]), presumably because its dissociation rate is much higher than its translocation rate. 4) The pH dependence of 1/tau shows that the dimeric species Cal2LiLi, Cal2NaNa, Cal2KH, and Cal2CsH are the dominant species translocating M+. The rate constant associated with their translocation has been estimated to be approximately 5 x 10(3) s-1. With this magnitude for the rate constants, the dimer dissociation constants of these species in the membrane have been estimated to be approximately 4, 1, 0.05, and 0.04 M, respectively. 5) Contrary to the claims made in the literature, the data obtained in the DeltapH decay studies do not favor the channel mechanism for the ion transport in this system. 6) However, they support the hypothesis that the dissociation of the divalent metal ion A23187 complex is the rate limiting step of A23187-mediated divalent metal ion transport. PMID- 9746515 TI - Inactivation of single cardiac Na+ channels in three different gating modes. AB - In small cell-attached patches containing one and only one Na+ channel, inactivation was studied in three different gating modes, namely, the fast inactivating F mode and the more slowly inactivating S mode and P mode with similar inactivation kinetics. In each of these modes, ensemble-averaged currents could be fitted with a Hodgkin-Huxley-type model with a single exponential for inactivation (tauh). tauh declined from 1.0 ms at -60 mV to 0.1 ms at 0 mV in the F mode, from 4.6 ms at -40 mV to 1.1 ms at 0 mV in the S mode, and from 4.5 ms at -40 mV to 0.8 ms at +20 mV in the P mode, respectively. The probability of non empty traces (net), the mean number of openings per non-empty trace (op/tr), and the mean open probability per trace (popen) were evaluated at 4-ms test pulses. net inclined from 30% at -60 mV to 63% at 0 mV in the F mode, from 4% at -90 mV to 90% at 0 mV in the S mode, and from 2% at -60 mV to 79% at +20 mV in the P mode. op/tr declined from 1.4 at -60 mV to 1.1 at 0 mV in the F mode, from 4.0 at -60 mV to 1.2 at 0 mV in the S mode, and from 2.9 at -40 mV to 1.6 at +20 mV in the P mode. popen was bell-shaped with a maximum of 5% at -30 mV in the F mode, 48% at -50 mV in the S mode, and 16% at 0 mV in the P mode. It is concluded that 1) a switch between F and S modes may reflect a functional change of inactivation, 2) a switch between S and P modes may reflect a functional change of activation, 3) tauh is mainly determined by the latency until the first channel opening in the F mode and by the number of reopenings in the S and P modes, 4) at least in the S and P modes, inactivation is independent of pore opening, and 5) in the S mode, mainly open channels inactivate, and in the P mode, mainly closed channels inactivate. PMID- 9746517 TI - Chloride channel blockers inhibit Ca2+ uptake by the smooth muscle sarcoplasmic reticulum. AB - Despite the fact that Ca2+ transport into the sarcoplasmic reticulum (SR) of muscle cells is electrogenic, a potential difference is not maintained across the SR membrane. To achieve electroneutrality, compensatory charge movement must occur during Ca2+ uptake. To examine the role of Cl- in this charge movement in smooth muscle cells, Ca2+ transport into the SR of saponin-permeabilized smooth muscle cells was measured in the presence of various Cl- channel blockers or when I-, Br-, or SO42- was substituted for Cl-. Calcium uptake was inhibited in a dose dependent manner by 5-nitro-2-(3-phenylpropylamino) benzoic acid (NPPB) and by indanyloxyacetic acid 94 (R(+)-IAA-94), but not by niflumic acid or 4,4' dinitrostilbene-2,2'-disulfonic acid (DNDS). Smooth muscle SR Ca2+ uptake was also partially inhibited by the substitution of SO42- for Cl-, but not when Cl- was replaced by I- or Br-. Neither NPPB nor R(+)-IAA-94 inhibited Ca2+ uptake into cardiac muscle SR vesicles at concentrations that maximally inhibited uptake in smooth muscle cells. These results indicate that Cl- movement is important for charge compensation in smooth muscle cells and that the Cl- channel or channels involved are different in smooth and cardiac muscle cells. PMID- 9746518 TI - Ca2+ removal mechanisms in rat cerebral resistance size arteries. AB - Tissue blood flow and blood pressure are each regulated by the contractile behavior of resistance artery smooth muscle. Vascular diseases such as hypertension have also been attributed to changes in vascular smooth muscle function as a consequence of altered Ca2+ removal. In the present study of Ca2+ removal mechanisms, in dissociated single cells from resistance arteries using fura-2 microfluorimetry and voltage clamp, Ca2+ uptake by the sarcoplasmic reticulum and extrusion by the Ca2+ pump in the cell membrane were demonstrably important in regulating Ca2+. In contrast, the Na+-Ca2+ exchanger played no detectable role in clearing Ca2+. Thus a voltage pulse to 0 mV, from a holding potential of -70 mV, triggered a Ca2+ influx and increased intracellular Ca2+ concentration ([Ca2+]i). On repolarization, [Ca2+]i returned to the resting level. The decline in [Ca2+]i consisted of three phases. Ca2+ removal was fast immediately after repolarization (first phase), then plateaued (second phase), and finally accelerated just before [Ca2+]i returned to resting levels (third phase). Thapsigargin or ryanodine, which each inhibit Ca2+ uptake into stores, did not affect the first but significantly inhibited the third phase. On the other hand, Na+ replacement with choline+ did not affect either the phasic features of Ca2+ removal or the absolute rate of its decline. Ca2+ removal was voltage-independent; holding the membrane potential at 120 mV, rather than at -70 mV, after the voltage pulse to 0 mV, did not attenuate Ca2+ removal rate. These results suggest that Ca2+ pumps in the sarcoplasmic reticulum and the plasma membrane, but not the Na+-Ca2+ exchanger, are important in Ca2+ removal in cerebral resistance artery cells. PMID- 9746520 TI - Membrane surface-charge titration probed by gramicidin A channel conductance. AB - We manipulate lipid bilayer surface charge and gauge its influence on gramicidin A channel conductance by two strategies: titration of the lipid charge through bulk solution pH and dilution of a charged lipid by neutral. Using diphytanoyl phosphatidylserine (PS) bilayers with CsCl aqueous solutions, we show that the effects of lipid charge titration on channel conductance are masked 1) by conductance saturation with Cs+ ions in the neutral pH range and 2) by increased proton concentration when the bathing solution pH is less than 3. A smeared charge model permits us to separate different contributions to the channel conductance and to introduce a new method for "bilayer pKa" determination. We use the Gouy-Chapman expression for the charged surface potential to obtain equilibria of protons and cations with lipid charges. To calculate cation concentration at the channel mouth, we compare different models for the ion distribution, exact and linearized forms of the planar Poisson-Boltzmann equation, as well as the construction of a "Gibbs dividing surface" between salt bath and charged membrane. All approximations yield the intrinsic pKain of PS lipid in 0.1 M CsCl to be in the range 2.5-3.0. By diluting PS surface charge at a fixed pH with admixed neutral diphytanoyl phosphatidylcholine (PC), we obtain a conductance decrease in magnitude greater than expected from the electrostatic model. This observation is in accord with the different conductance saturation values for PS and PC lipids reported earlier (, Biochim. Biophys. Acta. 552:369 378) and verified in the present work for solvent-free membranes. In addition to electrostatic effects of surface charge, gramicidin A channel conductance is also influenced by lipid-dependent structural factors. PMID- 9746519 TI - In situ characterization of the Ca2+ sensitivity of large conductance Ca2+ activated K+ channels: implications for their use as near-membrane Ca2+ indicators in smooth muscle cells. AB - The Ca2+ sensitivity of large conductance Ca2+- and voltage-activated K+ channels (BKV,Ca) has been determined in situ in freshly isolated myocytes from the guinea pig urinary bladder. In this study, in situ denotes that BKV,Ca channel activity was recorded without removing the channels from the cell. By combining patch clamp recording in the cell-attached configuration and microfluorometry of fura 2, we were able to correlate BKV,Ca channel activity with changes in cytoplasmic intracellular [Ca2+] ([Ca2+]i). The latter were induced by ionomycin, an electroneutral Ca2+ ionophore. At 0 mV, the Hill coefficient (nH) and the [Ca2+]i to attain half of the maximal BKV,Ca channel activity (Ca50) were 8 and 1 microM, respectively. The data suggest that this large Hill number was not a consequence of the difference between the near-membrane [Ca2+] ([Ca2+]s) and the bulk [Ca2+]i, indicated by fura-2. High Hill numbers in the activation by Ca2+ of BKV,Ca channels have been seen by different groups (e.g., filled squares in Fig. 4 of Silberberg, S. D., A. Lagrutta, J. P. Adelman, and K. L. Magleby. 1996. Biophys. J. 70:2640-2651). However, such high nH has always been considered a peculiarity rather than the rule. This work shows that a high Ca2+ cooperativity is the normal situation for BKV,Ca channels in myocytes from guinea pig urinary bladder. Furthermore, the Ca50 did not display any significant variation among different channels or cells. It was also evident that BKV,Ca channel activity could decrease in elevated [Ca2+]i, either partially or completely. This work implies that the complete activation of BKV,Ca channels occurs with a smaller increment in [Ca2+]s than previously expected from in vitro characterization of the Ca2+ sensitivity of these channels. Additionally, it appears that the activity of BKV,Ca channels in situ does not strictly follow changes in near membrane [Ca2+]. PMID- 9746521 TI - Basal activation of ATP-sensitive potassium channels in murine colonic smooth muscle cell. AB - The function and molecular expression of ATP-sensitive potassium (KATP) channels in murine colonic smooth muscle was investigated by intracellular electrical recording from intact muscles, patch-clamp techniques on isolated smooth muscle myocytes, and reverse transcription polymerase chain reaction (RT-PCR) on isolated cells. Lemakalim (1 microM) caused hyperpolarization of intact muscles (17. 2 +/- 3 mV). The hyperpolarization was blocked by glibenclamide (1-10 microM). Addition of glibenclamide (10 microM) alone resulted in membrane depolarization (9.3 +/- 1.7 mV). Lemakalim induced an outward current of 15 +/- 3 pA in isolated myocytes bathed in 5 mM external K+ solution. Application of lemakalim to cells in symmetrical K+ solutions (140/140 mM) resulted in a 97 +/- 5 pA inward current. Both currents were blocked by glibenclamide (1 microM). Pinacidil (1 microM) also activated an inwardly rectifying current that was insensitive to 4-aminopyridine and barium. In single-channel studies, lemakalim (1 microM) and diazoxide (300 microM) increased the open probability of a 27-pS K+ channel. Openings of these channels decreased with time after patch excision. Application of ADP (1 mM) or ATP (0.1 mM) to the inner surface of the patches reactivated channel openings. The conductance and characteristics of the channels activated by lemakalim were consistent with the properties of KATP. RT-PCR demonstrated the presence of Kir 6.2 and SUR2B transcripts in colonic smooth muscle cells; transcripts for Kir 6.1, SUR1, and SUR2A were not detected. These molecular studies are the first to identify the molecular components of KATP in colonic smooth muscle cells. Together with the electrophysiological experiments, we conclude that KATP channels are expressed in murine colonic smooth muscle cells and suggest that these channels may be involved in dual regulation of resting membrane potential, excitability, and contractility. PMID- 9746522 TI - Open channel block and alteration of N-methyl-D-aspartic acid receptor gating by an analog of phencyclidine. AB - We investigated inhibition of the N-methyl-D-aspartic acid (NMDA) receptor channel complex by N-ethyl-1,4,9, 9alpha-tetrahydro-4alphaR-cis-4alphaH-fluoren ++ +4alpha-amine (NEFA), a structural analog of phencyclidine (PCP). Using the whole-cell recording technique, we demonstrated that NEFA inhibits NMDA responses with an IC50 of 0.51 microM at -66 mV. We determined that NEFA binds to the open channel, and subsequently the channel can close and trap the blocker. Once the channel has closed, NEFA is unable to dissociate until the channel reopens. Single-channel recordings revealed that NEFA reduces the mean open time of single NMDA-activated channels in a concentration-dependent manner with a forward blocking rate (k+) of 39.9 microM-1 s-1. A computational model of antagonism by NEFA was developed and constrained using kinetic measurements of single-channel data. By multiple criteria, only models in which blocker binding in the channel causes a change in receptor operation adequately fit or predicted whole-cell data. By comparing model predictions and experimental measurements of NEFA action at a high NMDA concentration, we determined that NEFA affects receptor operation through an influence on channel gating. We conclude that inhibition of NMDA receptors by PCP-like blockers involves a modification of channel gating as well as block of current flow through the open channel. PMID- 9746523 TI - Epibatidine activates muscle acetylcholine receptors with unique site selectivity. AB - We recently showed that at desensitized muscle nicotinic receptors, epibatidine selects by 300-fold between the two agonist binding sites. To determine whether receptors in the resting, activatible state show similar site selectivity, we studied epibatidine-induced activation of mouse fetal and adult receptors expressed in 293 HEK cells. Kinetic analysis of single-channel currents reveals that (-)-epibatidine binds with 15-fold selectivity to sites of adult receptors and 75-fold selectivity to sites of fetal receptors. For each receptor subtype, site selectivity arises solely from different rates of epibatidine dissociation from the two sites. To determine the structural basis for epibatidine selectivity, we introduced mutations into either the gamma or the delta subunit and measured epibatidine binding and epibatidine-induced single-channel currents. Complexes formed by alpha and mutant gamma(K34S+F172I) subunits bind epibatidine with increased affinity compared to alphagamma complexes, whereas the kinetics of alpha2betadeltagamma(K34S+F172I) receptors reveal no change in affinity of the low-affinity site, but increased affinity of the high-affinity site. Conversely, complexes formed by alpha and mutant delta(S36K+I178F) subunits bind epibatidine with decreased affinity compared to alphadelta complexes, whereas the kinetics of alpha2betagammadelta(S36K+I178F) and alpha2betaepsilondelta(S36K+I178F) receptors show markedly reduced sensitivity to epibatidine. The overall data show that epibatidine activates muscle receptors by binding with high affinity to alphagamma and alphaepsilon sites, but with low affinity to the alphadelta site. PMID- 9746524 TI - Loss of shaker K channel conductance in 0 K+ solutions: role of the voltage sensor. AB - In potassium-free solutions some types of K channels enter a long-lasting nonconducting or "defunct" state. It is known that Shaker K channels must open in K+-free solutions to become defunct. Gating current studies presented here indicate an abnormal conformation in the defunct state that restricts S4 movement and alters its kinetics. Thus an abnormality initiated in the P region spreads to the gating apparatus. We find that channels most readily become defunct on repolarization to an intermediate voltage, thus prolonging occupancy of one of the several intermediate closed states. The state dependence of becoming defunct was further dissected by using the gating mutant L382A. Simply closing this channel at 0 mV (reversing the last activation step) does not make the mutant channel defunct. Instead, it is necessary to move further left (more fully closed) in the activation sequence. This was confirmed with ShIR experiments showing that channels become defunct only if there is inward gating charge movement. Rapid transit through the intermediate states, achieved at very negative voltage, is relatively ineffective at making channels defunct. Several mutations that removed C-type inactivation also made the channels resistant to becoming defunct. Our results show that normal gating current cannot be stably recorded in the absence of K+. PMID- 9746525 TI - Origin of reproducibility in the responses of retinal rods to single photons. AB - The single photon responses of retinal rod cells are remarkably reproducible, allowing the number and timing of photon absorptions to be encoded accurately. This reproducibility is surprising because the elementary response arises from a single rhodopsin molecule, and typically signals from single molecules display large intertrial variations. We have investigated the mechanisms that make the rod's elementary response reproducible. Our experiments indicate that reproducibility cannot be explained by saturation within the transduction cascade, by Ca2+ feedback, or by feedback control of rhodopsin shutoff by any known element of the cascade. We suggest instead that deactivation through a series of previously unidentified transitions allows the catalytic activity of a single rhodopsin molecule to decay with low variability. Two observations are consistent with this view. First, the time course of rhodopsin's catalytic activity could not be accounted for by the time required for the known steps in rhodopsin deactivation-phosphorylation and arrestin binding. Second, the variability of the elementary response increased when phosphorylation was made rate-limiting for rhodopsin shutoff. PMID- 9746526 TI - A new monofluorinated phosphatidylcholine forms interdigitated bilayers. AB - 16-Fluoropalmitic acid was synthesized from 16-hydroxypalmitic acid using diethylaminosulfur trifluoride. This monofluorinated fatty acid then was used to make 1-palmitoyl-2-[16-fluoropalmitoyl]-phosphatidylcholine (F-DPPC) as a fluorinated analog of dipalmitoylphosphatidylcholine (DPPC). Surprisingly, we found that the phase transition temperature (Tm) of F-DPPC occurs near 50 degrees C, approximately 10 degrees C higher than its nonfluorinated counterpart, DPPC, as judged by both differential scanning calorimetry and infrared spectroscopy. The pretransition observed for DPPC is absent in F-DPPC. A combination of REDOR, rotational-echo double-resonance, and conventional solid-state NMR experiments demonstrates that F-DPPC forms a fully interdigitated bilayer in the gel phase. Electron paramagnetic resonance experiments show that below Tm, the hydrocarbon chains of F-DPPC are more motionally restricted than those of DPPC. X-ray scattering experiments confirm that the thickness and packing of gel phase F-DPPC is similar to that of heptanetriol-induced interdigitated DPPC. F-DPPC is the first phosphoglyceride containing sn-1 and sn-2 ester-linked fatty acyl chains of equal length that spontaneously forms interdigitated bilayers in the gel state in the absence of inducing agents such as alcohols. PMID- 9746527 TI - Filipin-induced lesions in planar phospholipid bilayers imaged by atomic force microscopy. AB - Filipin is a macrolide polyene with antifungal activity belonging to the same family of antibiotics as amphotericin B and nystatin. Despite the spectroscopy and electron microscopy studies of its interaction with natural membranes and membrane model systems, several aspects of its biochemical action, such as the role of membrane sterols, remain to be completely understood. We have used atomic force microscopy (AFM) to study the effect of filipin on dipalmitoylphosphatidylethanolamine bilayers in the presence and absence of cholesterol. The bilayers were prepared by Langmuir-Blodgett deposition over mica and imaged under water. It was shown that filipin-induced lesions could only be found in membranes with cholesterol. In close agreement with electron microscopy results, we have reported the presence of densely packed circular protrusions in the membrane with a mean diameter of 19 nm (corrected for convolution with AFM tip) and 0.4 nm height. Larger circular protrusions (90 nm diameter and 2.5 nm height) and doughnut-shaped lesions were also detected. These results demonstrate that filipin-induced lesions in membranes previously observed by electron microscopy are not biased by artifacts resulting from sample preparation. Filipin aggregates in aqueous solution could also be imaged for the first time. These polydisperse spherical structures were observed in samples with and without cholesterol. PMID- 9746528 TI - Coupled plasmon-waveguide resonance spectroscopy studies of the cytochrome b6f/plastocyanin system in supported lipid bilayer membranes. AB - The incorporation of cytochrome (cyt) b6f into a solid-supported planar egg phosphatidylcholine (PC) bilayer membrane and complex formation with plastocyanin have been studied by a variant of surface plasmon resonance called coupled plasmon-waveguide resonance (CPWR) spectroscopy, developed in our laboratory. CPWR combines greatly enhanced sensitivity and spectral resolution with direct measurement of anisotropies in refractive index and optical extinction coefficient, and can therefore probe structural properties of lipid-protein and protein-protein interactions. Cyt b6f incorporation into the membrane proceeds in two stages. The first occurs at low protein concentration and is characterized by an increase in total proteolipid mass without significant changes in the molecular order of the system, as demonstrated by shifts of the resonance position to larger incident angles without changing the refractive index anisotropy. The second stage, occurring at higher protein concentrations, results in a decrease in both the mass density and the molecular order of the system, evidenced by shifts of the resonance position to smaller incident angles and a large decrease in the membrane refractive index anisotropy. Plastocyanin can bind to such a proteolipid system in three different ways. First, the addition of plastocyanin before the second stage of b6f incorporation begins results in complex formation between the two proteins with a KD of approximately 10 microM and induces structural changes in the membrane that are similar to those occurring during the second stage of complex incorporation. The addition of larger amounts of plastocyanin under these conditions leads to nonspecific binding to the lipid phase with a KD of approximately 180 microM. Finally, the addition of plastocyanin after the completion of the second phase of b6f incorporation results in tighter binding between the two proteins (KD approximately 1 microM). Quantitation of the binding stoichiometry indicates that two plastocyanin molecules bind tightly to the dimeric form of the cyt b6f complex, assuming random insertion of the cytochrome into the bilayer. The structural basis for these results and formation of the proteolipid membrane are discussed. PMID- 9746529 TI - Orientation dependence of displacements by a single one-headed myosin relative to the actin filament. AB - Displacements of single one-headed myosin molecules in a sparse myosin-rod cofilament were measured from bead displacements at various angles relative to an actin filament by dual optical trapping nanometry. The sparse myosin-rod cofilaments, 5-8 micron long, were synthesized by slowly mixing one-headed myosin prepared by papain digestion with myosin rods at molar ratios of 1:400 to 1:1500, so that one to four one-headed myosin molecules were on average scattered along the cofilament. The bead displacement was approximately 10 nm at low loads ( approximately 0.5 pN) and at angles of 5-10 degrees between the actin and myosin filaments (near physiologically correct orientation). The bead displacement decreased with an increase in the angle. The bead displacement at nearly 90 degrees was approximately 0 nm. When the angle was increased to approximately 150 degrees-170 degrees, the bead displacements increased to 5 nm. A native two headed myosin showed similar size and orientation dependence of bead displacements as a one-headed myosin. PMID- 9746530 TI - Calcium transients and the effect of a photolytically released calcium chelator during electrically induced contractions in rabbit rectococcygeus smooth muscle. AB - Intracellular Ca2+ was determined with the fura-2 technique during electrically induced contractions in the rabbit rectococcygeus smooth muscle at 22 degreesC. The muscles were electrically activated to give short, reproducible contractions. Intracellular [Ca2+] increased during activation; the increase in [Ca2+] preceded force development by approximately 2 s. After cessation of stimulation Ca2+ fell, preceding the fall in force by approximately 4 s. The fluorescence properties of fura-2 were determined with time-resolved spectroscopy using synchrotron light at the MAX-storage ring, Lund, Sweden. The fluorescence decay of free fura-2 was best described by two exponential decays (time constants approximately 0.5 and 1.5 ns) at low Ca2+ (pCa 9). At high Ca2+ (pCa 4.5), fluorescence decay became slower and could be fitted by one exponential decay (1.9 ns). Time-resolved anisotropy of free fura-2 was characteristic of free rotational motion (correlation time 0.3 ns). Motion of fura-2 could be markedly inhibited by high concentrations of creatine kinase. Time-resolved spectroscopy measurements of muscle fibers loaded with fura-2 showed that the fluorescence lifetime of the probe was longer, suggesting an influence of the chemical environment. Anisotropy measurements revealed, however, that the probe was mobile in the cells. The Ca2+ dependence of contraction and relaxation was studied using a photolabile calcium chelator, diazo-2, which could be loaded into the muscle cells in a similar manner as fura-2. Photolysis of diazo-2 leads to an increase in its Ca2+-affinity and a fall in free Ca2+. When muscles that had been loaded with diazo-2 were illuminated with UV light flashes during the rising phase of contraction, the rate of contraction became slower, suggesting a close relation between intracellular Ca2+ and the cross-bridge interaction. In contrast, photolysis during relaxation did not influence the rate of force decay, suggesting that relaxation of these contractions is not determined by the rate of Ca2+ removal or due to an increased Ca2+ sensitivity, but instead is limited by other processes such as deactivation by dephosphorylation or detachment of tension-bearing cross bridges, possibly regulated by thin filament systems. PMID- 9746531 TI - Regulation of catch muscle by twitchin phosphorylation: effects on force, ATPase, and shortening. AB - Recent experiments on permeabilized anterior byssus retractor muscle (ABRM) of Mytilus edulis have shown that phosphorylation of twitchin releases catch force at pCa > 8 and decreases force at suprabasal but submaximum [Ca2+]. Twitchin phosphorylation decreases force with no detectable change in ATPase activity, and thus increases the energy cost of force maintenance at subsaturating [Ca2+]. Similarly, twitchin phosphorylation causes no change in unloaded shortening velocity (Vo) at any [Ca2+], but when compared at equal submaximum forces, there is a higher Vo when twitchin is phosphorylated. During calcium activation, the force-maintaining structure controlled by twitchin phosphorylation adjusts to a 30% Lo release to maintain force at the shorter length. The data suggest that during both catch and calcium-mediated submaximum contractions, twitchin phosphorylation removes a structure that maintains force with a very low ATPase, but which can slowly cycle during submaximum calcium activation. A quantitative cross-bridge model of catch is presented that is based on modifications of the Hai and Murphy (1988. Am. J. Physiol. 254:C99-C106) latch bridge model for regulation of mammalian smooth muscle. PMID- 9746532 TI - Cholesterol distribution in living cells: fluorescence imaging using dehydroergosterol as a fluorescent cholesterol analog. AB - Cholesterol is an important constituent of most mammalian cell membranes and its concentration in various cellular membranes is tightly regulated. Although there is much information about cholesterol distribution and trafficking in cells, it is primarily derived from indirect measurements, and the results obtained using different approaches are often conflicting. A cholesterol analog that faithfully mimics the properties of cholesterol and can be followed in living cells would thus be very useful. In this study, we report the fluorescence imaging of such an analog, dehydroergosterol (DHE), in living cells. DHE differs from cholesterol in having three additional double bonds and an extra methyl group. In model systems, DHE closely mimics the behavior of native cholesterol. Using triple-labeling studies, we show that DHE colocalizes extensively with endocytosed transferrin, an endocytic recycling compartment marker, and with a marker for the trans-Golgi network, Tac-TGN38. This distribution of DHE is qualitatively similar to that observed when cells are labeled with the fluorescent cholesterol-binding polyene antibiotic, filipin, although there are differences in apparent proportions of DHE and filipin that are localized at the plasma membrane. Another cholesterol derivative, 25-NBD-cholesterol, has a structure that is compromised by the presence of a bulky NBD group and does not distribute to the same organelles as DHE or filipin. In addition, we show in this manuscript that kinetic processes can be followed in living cells by monitoring recovery of DHE fluorescence in a photobleached region over time. Our observations provide evidence for the presence of a large intracellular cholesterol pool in the endocytic recycling compartment and the trans-Golgi network that might play important roles in the trafficking of lipids, lipid-anchored proteins, and transmembrane proteins that preferentially partition into cholesterol-enriched membrane domains. In addition, this intracellular cholesterol pool might be involved in the maintenance of cellular cholesterol homeostasis. PMID- 9746533 TI - MgADP promotes a catch-like state developed through force-calcium hysteresis in tonic smooth muscle. AB - Tonic rabbit femoral artery and phasic rabbit ileum smooth muscles permeabilized with Triton X-100 were activated either by increasing [Ca2+] from pCa > 8.0 to pCa 6.0 (calcium-ascending protocol) or contracted at pCa 6.0 before lowering [Ca2+] (calcium-descending protocol). The effects of, respectively, high [MgATP]/low [MgADP] [10 mM MgATP + creatine phosphate (CP) + creatine kinase (CK)] or low [MgATP]/[MgADP] (2 mM MgATP, 0 CP, 0 CK) on the "force-[Ca]" relationships were determined. In femoral artery at low, but not at high, [MgATP]/[MgADP] the force and the ratio of stiffness/force at pCa 7.2 were significantly higher under the calcium-descending than calcium-ascending protocols (54% vs. 3% of Po, the force at pCa 6.0) (force hysteresis); the levels of regulatory myosin light chain (MLC20) phosphorylation (9 +/- 2% vs. 10 +/- 2%) and the velocities of unloaded shortening V0 (0.02 +/- 0.004 l/s with both protocols) were not significantly different. No significant force hysteresis was detected in rabbit ileum under either of these experimental conditions. [MgADP], measured in extracts of permeabilized femoral artery strips by two methods, was 130-140 microM during maintained force under the calcium-descending protocol. Exogenous CP (10 mM) applied during the descending protocol reduced endogenous [MgADP] to 46 +/- 10 microM and abolished force hysteresis: residual force at low [Ca2+] was 17 +/- 5% of maximal force. We conclude that the proportion of force generating nonphosphorylated (AMdp) relative to phosphorylated cross-bridges is higher on the Ca2+-descending than on the Ca2+-ascending force curve in tonic smooth muscle, that this population of positively strained dephosphorylated cross bridges has a high affinity for MgADP, and that the dephosphorylated AMdp . MgADP state makes a significant contribution to force maintenance at low levels of MLC20 phosphorylation. PMID- 9746534 TI - Geometrical and sequence characteristics of alpha-helices in globular proteins. AB - Understanding the sequence-structure relationships in globular proteins is important for reliable protein structure prediction and de novo design. Using a database of 1131 alpha-helices with nonidentical sequences from 205 nonhomologous globular protein chains, we have analyzed structural and sequence characteristics of alpha-helices. We find that geometries of more than 99% of all the alpha helices can be simply characterised as being linear, curved, or kinked. Only a small number of alpha-helices ( approximately 4%) show sharp localized bends in their middle regions, and thus are classified as kinked. Approximately three fourths (approximately 73%) of the alpha-helices in globular proteins show varying degrees of smooth curvature, with a mean radius of curvature of 65 +/- 33 A; longer helices are less curved. Computation of helix accessibility to the solvent indicates that nearly two-thirds of the helices ( approximately 66%) are largely buried in the protein core, and the length and geometry of the helices are not correlated with their location in the protein globule. However, the amino acid compositions and propensities of individual amino acids to occur in alpha helices vary with their location in the protein globule, their geometries, and their lengths. In particular, Gln, Glu, Lys, and Arg are found more often in helices near the surface of globular proteins. Interestingly, kinks often seem to occur in regions where amino acids with low helix propensities (e.g., beta branched and aromatic residues) cluster together, in addition to those associated with the occurrence of proline residues. Hence the propensities of individual amino acids to occur in a given secondary structure depend not only on conformation but also on its length, geometry, and location in the protein globule. PMID- 9746535 TI - Thermal motions in bacteriorhodopsin at different hydration levels studied by neutron scattering: correlation with kinetics and light-induced conformational changes. AB - Bacteriorhodopsin (BR) is a transmembrane protein in the purple membrane (PM) of Halobacterium salinarum. Its function as a light-driven proton pump is associated with a cycle of photointermediates which is strongly hydration-dependent. Using energy-resolved neutron scattering, we analyzed the thermal motions (in the nanosecond-to-picosecond time range) in PM at different hydration levels. Two main populations of motions were found that responded differently to water binding. Striking correlations appeared between these "fast" motions and the "slower" kinetic constants (in the millisecond time range) of relaxations and conformational changes occurring during the photocycle. PMID- 9746536 TI - The active site structure of tetanus neurotoxin resolved by multiple scattering analysis in X-Ray absorption spectroscopy. AB - A detailed study of the x-ray absorption spectrum of tetanus neurotoxin in the K edge EXAFS region of the zinc absorber is presented that allows the complete identification of the amino acid residues coordinated to the zinc active site. A very satisfactory interpretation of the experimental data can be given if multiple scattering contributions are included in the analysis. Comparing the absorption spectrum of tetanus neurotoxin to that of two other structurally similar zinc-endopeptidases, thermolysin and astacin, in which the zinc coordination mode is known from crystallographic data, we conclude that in tetanus neurotoxin, besides a water molecule, zinc is coordinated to two histidines and a tyrosine. PMID- 9746537 TI - Primary sequence and solution conformation of ferrocytochrome c-552 from Nitrosomonas europaea. AB - Cytochrome c-552 from Nitrosomonas europaea is a 9.1-kDa monoheme protein that is a member of the bacterial cytochrome c-551 family. The gene encoding for c-552 has been cloned and sequenced and the primary sequence of the product deduced. Proton resonance assignments were made for all main-chain and most side-chain protons in the diamagnetic, reduced form by two-dimensional NMR techniques. Distance constraints (1056) were determined from nuclear Overhauser enhancements, and torsion angle constraints (88) were determined from scalar coupling estimates. Solution conformations for the protein were computed by the hybrid distance geometry-simulated annealing approach. For 20 computed structures, the root mean squared deviation from the average position of equivalent atoms was 0.84 A (sigma = 0.12) for backbone atoms over all residues. Analysis by residue revealed there were three regions clearly less well defined than the rest of the protein: the first two residues at the N-terminus, the last two at the C terminus, and a loop region from residues 34 to 40. Omitting these regions from the comparison, the root mean squared deviation was 0.61 A (sigma = 0.13) for backbone atoms, 0.86 A (sigma = 0.12) for all associated heavy atoms, and 0. 43 A (sigma = 0.17) for the heme group. The global folding of the protein is consistent with others in the c-551 family. A deletion at the N-terminus relative to other family members had no impact on the global folding, whereas an insertion at residue 65 did affect the way the polypeptide packs against the methionine ligated side of the heme. The effects of specific substitutions will be discussed. The structure of c-552 serves to delineate essential features of the c 551 family. PMID- 9746538 TI - Cl- regulates the structure of the fibrin clot. AB - The differences between coarse and fine fibrin clots first reported by Ferry have been interpreted in terms of nonspecific ionic strength effects for nearly 50 years and have fostered the notion that fibrin polymerization is largely controlled by electrostatic forces. Here we report spectroscopic and electron microscopy studies carried out in the presence of different salts that demonstrate that this long-held interpretation needs to be modified. In fact, the differences are due entirely to the specific binding of Cl- to fibrin fibers and not to generic ionic strength or electrostatic effects. Binding of Cl- opposes the lateral aggregation of protofibrils and results in thinner fibers that are also more curved than those grown in the presence of inert anions such as F-. The effect of Cl- is pH dependent and increases at pH > 8.0, whereas fibers grown in the presence of F- remain thick over the entire pH range from 6.5 to 9.0. From the pH dependence of the Cl- effect it is suggested that the anion exerts its role by increasing the pKa of a basic group ionizing around pH 9.2. The important role of Cl- in structuring the fibrin clot also clarifies the role played by the release of fibrinopeptide B, which leads to slightly thicker fibers in the presence of Cl- but actually reduces the size of the fibers in the presence of F . This effect becomes more evident at high, close to physiological concentrations of fibrinogen. We conclude that Cl- is a basic physiological modulator of fibrin polymerization and acts to prevent the growth of thicker, stiffer, and straighter fibers by increasing the pKa of a basic group. This discovery opens new possibilities for the design of molecules that can specifically modify the clot structure by targeting the structural domains responsible for Cl- binding to fibrin. PMID- 9746539 TI - Nucleotide and Mg2+ dependency of the thermal denaturation of mitochondrial F1 ATPase. AB - The influence of adenine nucleotides and Mg2+ on the thermal denaturation of mitochondrial F1-ATPase (MF1) was analyzed. Differential scanning calorimetry in combination with ATPase activity experiments revealed the thermal unfolding of MF1 as an irreversible and kinetically controlled process. Three significant elements were analyzed during the thermal denaturation process: the endothermic calorimetric transition, the loss of ATP hydrolysis activity, and the release of tightly bound nucleotides. All three processes occur in the same temperature range, over a wide variety of conditions. The purified F1-ATPase, which contains three tightly bound nucleotides, denatures at a transition temperature (Tm) of 55 degrees C. The nucleotide and Mg2+ content of MF1 strongly influence the thermal denaturation process. First, further binding of nucleotides and/or Mg2+ to MF1 increases the thermal denaturation temperature, whereas the thermal stability of the enzyme is decreased upon removal of the endogenous nucleotides. Second, the stabilizing effect induced by nucleotides is smaller after hydrolysis of ATP (i.e., in the presence of ADP . Mg2+) than under nonhydrolytical conditions (i.e., absence of Mg2+ or using the nonhydrolyzable analog 5'-adenylyl imidodiphosphate). Third, whereas the thermal denaturation of MF1 fully loaded with nucleotides follows an apparent two-state kinetic process, denaturation of MF1 with a low nucleotide content follows more complex kinetics. Nucleotide content is therefore an important factor in determining the thermal stability of the MF1 complex, probably by strengthening existing intersubunit interactions or by establishing new ones. PMID- 9746541 TI - A DNA self-assembled monolayer for the specific attachment of unmodified double- or single-stranded DNA. AB - A novel method for DNA surface immobilization and a paradigm for the attachment of unmodified DNA of any length or sequence are described herein. The development of a DNA self-assembled monolayer (DNA-SAM) that incorporates a DNA-thiol into a monolayer of inert alkane thiolates is reported. This DNA-SAM specifically hybridized complementary oligonucleotides while resisting the nonspecific adsorption of noncomplementary DNA and irrelevant proteins. Duplex DNA, having a single-stranded "capture tail," specifically bound to the DNA-SAM if the sequence of the "tail" was complementary to DNA presented in the SAM. The sense strand of the hybridized duplex DNA could be covalently attached to the surface by an enzymatic ligation reaction (leaving the anti-sense strand dissociable). DNA binding proteins specifically bound to these surfaces only if their cognate sites were present in the duplex DNA. PMID- 9746540 TI - The use of variable density self-assembled monolayers to probe the structure of a target molecule. AB - VP16, a protein encoded by herpes simplex virus, has a well-characterized 78 amino acid acidic activation domain. When tethered to DNA, tandem repeats of an eight amino acid motif taken from this region stimulate the transcription of a nearby gene. This work addresses how these minimal activation motifs interact with a putative target, the general transcription factor TATA box binding protein (TBP), and the biological relevance of this mechanism of action. I developed novel biophysical techniques to discriminate among three possible mechanistic models that describe how reiterated peptide motifs could synergistically effect transcription: 1) the peptide motifs simultaneously bind to quasi-identical sites on TBP, producing a high-affinity bivalent interaction that holds the general transcription factor near the start site of transcription; 2) the binding of one recognition motif causes an allosteric effect that enhances the subsequent binding of additional peptide motifs; or 3) a high-affinity interaction between the peptide repeats and TBP does occur, but rather than being the result of a "bivalent" interaction, it results from the summation of multiple interactions between the target protein and the entire length of the peptide. I generated self assembled monolayers (SAMs) that presented different densities of the activation motif peptide in a two-dimensional array to test for avidity effects. Surface plasmon resonance (SPR) was used to measure the amount of target (TBP) binding as a function of the peptide density; a marked increase in avidity above a characteristic, critical peptide surface density was found. Competitive inhibition experiments were performed to compare the avidity of peptide motifs, tandemly repeated two or four times, and single motifs separated by a flexible linker. Four iterations of the motif, preincubated with TBP, inhibited its binding to high-density peptide surfaces approximately 250-fold better than two iterations. Single peptide motifs joined by a flexible amino acid linker inhibited TBP binding to surface peptide nearly as well as four tandem repeats. The results favor mechanistic model 1: reiterated activation motifs interact with TBP through a high-affinity interaction that is the result of the cooperative effect of single motifs simultaneously binding to separate sites on TBP. This finding is consistent with the idea that DNA-bound activation domains trigger the transcription of a nearby gene by tethering the general transcription factor, TBP, near the start site of transcription. PMID- 9746542 TI - Role of mitochondria in calcium regulation of spontaneously contracting cardiac muscle cells. AB - Mitochondrial involvement in the regulation of cytosolic calcium concentration ([Ca2+]i) in cardiac myocytes has been largely discounted by many authors. However, recent evidence, including the results of this study, has forced a reappraisal of this role. [Ca2+]i and Ca2+ in the mitochondria ([Ca2+]m) were measured in this study with specific fluorescent probes, fluo-3 and di-hydro-rhod 2, respectively; mitochondrial membrane potential (DeltaPsim) was monitored with JC-1. Addition of uncouplers or inhibitors of the mitochondrial respiratory chain was found to cause a twofold decrease in the rate of removal of Ca2+ from the cytosol after a spontaneously generated Ca2+ wave. These agents also caused a progressive elevation of [Ca2+]i, an increase in the number of hotspots of Ca2+ release (Ca2+ sparks), and depression of mitochondrial potential. The Ca2+ indicative fluorophore dihydro-rhod-2 has a net positive charge that contributes to selective accumulation by mitochondria, as supported by its co-localization with other mitochondrial-specific probes (MitoTracker Green). Treatment of dihydro-rhod-2-loaded cells with NaCN resulted in rapid formation of "black holes" in the otherwise uniformly banded pattern. These are likely to represent individual or small groups of mitochondria that have depressed mitochondrial potential, or have lost accumulated rhod-2 and/or Ca2+; all of these eventualities are possible upon onset of the mitochondrial permeability transition. Release of Ca2+ from the sarcoplasmic reticulum and the resultant spontaneous contractility of cardiac muscle are proposed to be triggered by the induction of the mitochondrial permeability transition and the subsequent loss of [Ca2+]m. PMID- 9746543 TI - Multiphoton excitation provides optical sections from deeper within scattering specimens than confocal imaging. AB - Multiphoton excitation fluorescence imaging generates an optical section of sample by restricting fluorophore excitation to the plane of focus. High photon densities, achieved only in the focal volume of the objective, are sufficient to excite the fluorescent probe molecules by density-dependent, multiphoton excitation processes. We present comparisons of confocal with multiphoton excitation imaging of identical optical sections within a sample. These side-by side comparisons of imaging modes demonstrate a significant advantage of multiphoton imaging; data can be obtained from deeper within biological specimens. Observations on a variety of biological samples showed that in all cases there was at least a twofold improvement in the imaging penetration depth obtained with multiphoton excitation relative to confocal imaging. The more pronounced degradation in image contrast deep within a confocally imaged sample is primarily due to scattered emission photons, which reduce the signal and increase the local background as measurements of point spread functions indicated that resolution does not significantly change with increasing depth for either mode of microscopy. Multiphoton imaging does not suffer from degradation of signal-to-background to nearly the same extent as confocal imaging because this method is insensitive to scatter of the emitted signal. Direct detection of emitted photons using an external photodetector mounted close to the objective (possible only in a multiphoton imaging system) improves system sensitivity and the utilization of scattered emission photons for imaging. We demonstrate that this technique provides yet further improvements in the capability of multiphoton excitation imaging to produce good quality images from deeper within tissue relative to confocal imaging. PMID- 9746544 TI - A simple method for high temporal resolution calcium imaging with dual excitation dyes. AB - Calcium-sensitive dual excitation dyes, such as fura-2, are now widely used to measure the free calcium concentration ([Ca2+]) in living cells. Preferentially, [Ca2+] is calculated in a ratiometric manner, but if calcium images need to be acquired at high temporal resolution, a potential drawback of ratiometry is that it requires equally fast switching of the excitation light between two wavelengths. To circumvent continuous excitation switching, some investigators have devised methods for calculating [Ca2+] from single-wavelength measurements combined with the acquisition of a single ratiometric pair of fluorescence images at the start of the recording. These methods, however, are based on the assumption that the concentration of the dye does not change during the experiment, a condition that is often not fulfilled. We describe here a method of single-wavelength calcium imaging, in which the dye concentration is estimated from ratiometric fluorescence image pairs acquired at regular intervals during the recording period, that furthermore includes a correction for the changing dye concentration in the calculation of [Ca2+]. PMID- 9746545 TI - Tapping mode atomic force microscopy of hyaluronan: extended and intramolecularly interacting chains. AB - The extracellular matrix polysaccharide hyaluronan has been examined by tapping mode atomic force microscopy. High molecular weight hyaluronan was deposited on mica from dilute aqueous solution and imaged in air. Long unbranched chains could be observed and were found to be compatible with the known covalent structure of hyaluronan. In addition, chains with evidence of intramolecular association were observed. In the simplest cases, the association took the form of loops stabilized by antiparallel double-stranded (probably double-helical) segments. In other cases, the polarity of the associated regions could not be determined. Extensive intramolecular association in long hyaluronan chains resulted in a fenestrated structure of the same type as that formed by intermolecular association at higher concentrations. PMID- 9746546 TI - Local measurements of viscoelastic parameters of adherent cell surfaces by magnetic bead microrheometry. AB - A magnetic bead microrheometer has been designed which allows the generation of forces up to 10(4) pN on 4.5 micron paramagnetic beads. It is applied to measure local viscoelastic properties of the surface of adhering fibroblasts. Creep response and relaxation curves evoked by tangential force pulses of 500-2500 pN (and approximately 1 s duration) on the magnetic beads fixed to the integrin receptors of the cell membrane are recorded by particle tracking. Linear three phasic creep responses consisting of an elastic deflection, a stress relaxation, and a viscous flow are established. The viscoelastic response curves are analyzed in terms of a series arrangement of a dashpot and a Voigt body, which allows characterization of the viscoelastic behavior of the adhering cell surface in terms of three parameters: an effective elastic constant, a viscosity, and a relaxation time. The displacement field generated by the local tangential forces on the cell surface is visualized by observing the induced motion of assemblies of nonmagnetic colloidal probes fixed to the membrane. It is found that the displacement field decays rapidly with the distance from the magnetic bead. A cutoff radius of Rc approximately 7 micron of the screened elastic field is established. Partial penetration of the shear field into the cytoplasm is established by observing the induced deflection of intracellular compartments. The cell membrane was modeled as a thin elastic plate of shear modulus mu * coupled to a viscoelastic layer, which is fixed to a solid support on the opposite side; the former accounts for the membrane/actin cortex, and the latter for the contribution of the cytoskeleton to the deformation of the cell envelope. It is characterized by the coupling constant chi characterizing the elasticity of the cytoskeleton. The coupling constant chi and the surface shear modulus mu * are obtained from the measured displacements of the magnetic and nonmagnetic beads. By analyzing the experimental data in terms of this model a surface shear modulus of mu * approximately 2 . 10(-3) Pa m to 4 . 10(-3) Pa m is found. By assuming an approximate plate thickness of 0.1 micron one estimates an average bulk shear modulus of mu approximately (2 / 4) . 10(-4) Pa, which is in reasonable agreement with data obtained by atomic force microscopy. The viscosity of the dashpot is related to the apparent viscosity of the cytoplasm, which is obtained by assuming that the top membrane is coupled to the bottom (fixed) membrane by a viscous medium. By application of the theory of diffusion of membrane proteins in supported membranes we find a coefficient of friction of bc approximately 2 . 10(9) Pa s/m corresponding to a cytoplasmic viscosity of 2 . 10(3) Pa s. PMID- 9746547 TI - Time course of the initial [Ca2+]i response to extracellular ATP in smooth muscle depends on [Ca2+]e and ATP concentration. AB - In response to extracellular application of 50 microM ATP, all individual porcine aortic smooth muscle cells respond with rapid rises from basal [Ca2+]i to peak [Ca2+]i within 5 s. The time from stimulus to the peak of the [Ca2+]i response increases with decreasing concentration of ATP. At ATP concentrations of 0.5 microM and below, the time to the [Ca2+]i peak varies more significantly from cell to cell than at higher concentrations, and each cell shows complicated initiation and decay kinetics. For any individual cell, the lag phase before a response decreases with increasing concentration of ATP. An increase in lag time with decreasing ATP concentration is also observed in the absence of extracellular Ca2+, but the lag phase is more pronounced, especially at concentrations of ATP below 0.5 microM. Whole-cell patch-clamp electrophysiology shows that in porcine aortic smooth muscle cells, ATP stimulates an inward current carried mainly by Cl- ion efflux with a time course similar to the [Ca2+]i changes and no detectable current from an ATP-gated cation channel. A simple signal cascade initiation kinetics model, starting with nucleotide receptor activation leading to IP3-mediated Ca2+ release from IP3-sensitive internal stores, fits the data and suggests that the kinetics of the Ca2+ response are dominated by upstream signal cascade components. PMID- 9746548 TI - How microtubules get fluorescent speckles. AB - The dynamics of microtubules in living cells can be seen by fluorescence microscopy when fluorescently labeled tubulin is microinjected into cells, mixing with the cellular tubulin pool and incorporating into microtubules. The subsequent fluorescence distribution along microtubules can appear "speckled" in high-resolution images obtained with a cooled CCD camera (Waterman-Storer and Salmon, 1997. J. Cell Biol. 139:417-434). In this paper we investigate the origins of these fluorescent speckles. In vivo microtubules exhibited a random pattern of speckles for different microtubules and different regions of an individual microtubule. The speckle pattern changed only after microtubule shortening and regrowth. Microtubules assembled from mixtures of labeled and unlabeled pure tubulin in vitro also exhibited fluorescent speckles, demonstrating that cellular factors or organelles do not contribute to the speckle pattern. Speckle contrast (measured as the standard deviation of fluorescence intensity along the microtubule divided by the mean fluorescence intensity) decreased as the fraction of labeled tubulin increased, and it was not altered by the binding of purified brain microtubule-associated proteins. Computer simulation of microtubule assembly with labeled and unlabeled tubulin showed that the speckle patterns can be explained solely by the stochastic nature of tubulin dimer association with a growing end. Speckle patterns can provide fiduciary marks in the microtubule lattice for motility studies or can be used to determine the fraction of labeled tubulin microinjected into living cells. PMID- 9746549 TI - Simultaneous measurements of actin filament turnover, filament fraction, and monomer diffusion in endothelial cells. AB - The analogous techniques of photoactivation of fluorescence (PAF) and fluorescence recovery after photobleaching (FRAP) have been applied previously to the study of actin dynamics in living cells. Traditionally, separate experiments estimate the mobility of actin monomer or the lifetime of actin filaments. A mathematical description of the dynamics of the actin cytoskeleton, however, predicts that the evolution of fluorescence in PAF and FRAP experiments depends simultaneously on the diffusion coefficient of actin monomer, D, the fraction of actin in filaments, FF, and the lifetime of actin filaments, tau (, Biophys. J. 69:1674-1682). Here we report the application of this mathematical model to the interpretation of PAF and FRAP experiments in subconfluent bovine aortic endothelial cells (BAECs). The following parameters apply for actin in the bulk cytoskeleton of subconfluent BAECs. PAF: D = 3.1 +/- 0.4 x 10(-8) cm2/s, FF = 0.36 +/- 0.04, tau = 7.5 +/- 2.0 min. FRAP: D = 5.8 +/- 1.2 x 10(-8) cm2/s, FF = 0.5 +/- 0.04, tau = 4.8 +/- 0.97 min. Differences in the parameters are attributed to differences in the actin derivatives employed in the two studies and not to inherent differences in the PAF and FRAP techniques. Control experiments confirm the modeling assumption that the evolution of fluorescence is dominated by the diffusion of actin monomer, and the cyclic turnover of actin filaments, but not by filament diffusion. The work establishes the dynamic state of actin in subconfluent endothelial cells and provides an improved framework for future applications of PAF and FRAP. PMID- 9746550 TI - Characterization of the sperm-induced calcium wave in Xenopus eggs using confocal microscopy. AB - We have used confocal microscopy to examine the [Ca2+]i increase in the albino eggs of the frog Xenopus laevis after fertilization. Eggs were placed in agar wells with their animal poles downward so that fertilization occurred preferentially in the equatorial plane, and confocal microscopy was used to provide a two-dimensional optical section through the three-dimensional Ca2+ wave. These data indicate that the wave of increased [Ca2+]i traverses the entire egg and converges uniformly on the antipode. We show that ratioing two different fluorescent dyes to correct for variations in cell thickness is not a reliable technique for this very thick cell due to differential absorption with depth. Indo-1-dextran proves to be a more reliable Ca2+ indicator in this respect. Indo 1-dextran measurements indicate that the resting [Ca2+]i is not uniform throughout the egg but exhibits a 15% higher [Ca2+]i in the cortex than deep in the cytoplasm. This difference is accentuated during wave propagation and is not dependent on extracellular Ca2+. The average peak [Ca2+]i in the center of the egg as the wave propagates through it is 0.7 microM, approximately 60% of the peak cortical [Ca2+]i. The wave velocity through the center of the egg (5.7 micron/s) is slower than that in the cortex (8.9 micron/s), and both velocities vary slightly during transit. The cortical wave speed is particularly high at the beginning (15.7 micron/s) and end (17.2 micron/s) of the wave. Eggs injected with 30-80 microM of 3 kD heparin to compete with inositol-1,4,5,-trisphosphate for binding to its receptor exhibited multiple localized spots of elevated [Ca2+]i, and many of these did not initiate a wave. For those that did lead to a wave, it was usually slow moving and exhibited a reduced (60% reduction) amplitude compared with controls. PMID- 9746551 TI - Simulation of the fertilization Ca2+ wave in Xenopus laevis eggs. AB - In the preceding paper Fontanilla and Nuccitelli (Biophysical Journal 75:2079 2087 (1998)) present detailed measurements of the shape and speed of the fertilization Ca2+ wave in Xenopus laevis eggs. In order to help interpret their results, we develop here a computational technique based on the finite element method that allows us to carry out realistic simulations of the fertilization wave. Our simulations support the hypothesis that the physiological state of the mature egg is bistable, i.e., that its cytoplasm can accommodate two alternative physiological Ca2+ concentrations: a low concentration characteristic of the prefertilization state and a greatly elevated concentration characteristic of the state following the passage of the wave. We explore this hypothesis by assuming that the bistability is due to the release and re-uptake properties of the endoplasmic reticulum (ER) as determined by inositol trisphosphate (IP3) receptor/Ca2+ channels and sarcoendoplasmic reticulum calcium ATPase (SERCA) pumps. When combined with buffered diffusion of Ca2+ in the cytoplasm, our simulations show that inhomogeneities in the Ca2+ release properties near the plasma membrane are required to explain the temporal and spatial dependences of the shape and speed of these waves. Our results are consistent with an elevated IP3 concentration near the plasma membrane in the unfertilized egg that is augmented significantly near the site of fertilization. These gradients are essential in determining the concave shape of the Ca2+ fertilization wave front. PMID- 9746552 TI - Supercharging accelerates T-tubule membrane potential changes in voltage clamped frog skeletal muscle fibers. AB - In voltage-clamp studies of single frog skeletal muscle fibers stained with the potentiometric indicator 1-(3-sulfonatopropyl)-4-[beta[2-(di-n-octylamino)-6 naphthyl] vinyl]pyridinium betaine (di-8 ANEPPS), fluorescence transients were recorded in response to both supercharging and step command pulses. Several illumination paradigms were utilized to study global and localized regions of the transverse tubule system (T-system). The rising phases of transients obtained from global illumination regions showed distinct accelerations when supercharging pulses were applied (95% of steady-state fluorescence achieved in 1.5 ms with supercharging pulses versus 14.6 ms with step pulses). When local transients were recorded at the edge of the muscle fiber, their kinetics resembled those of the applied waveform, but a similar relationship was not observed in transients from regions near the edge chosen to minimize the surface membrane contribution. We developed a model of the T-system capable of simulating membrane potential changes as a function of time and distance along the T-system cable and the associated fluorescence changes in regions corresponding to the experimental illumination strategies. A critical parameter was the access resistance term, for which values of 110-150 Omega.cm2 were adequate to fit the data. The results suggest that the primary mechanism through which supercharging pulses boost the kinetics of T-system voltage changes most likely involves their compensating the voltage attenuation across the access resistance at the mouth of the T-tubule. PMID- 9746554 TI - Antigenic determinants of Staphylococcus aureus type 5 and type 8 capsular polysaccharide vaccines. AB - Bacterial capsular polysaccharides (CP) are carbohydrate polymers comprised of repeating saccharide units. Several of these CP have side chains attached to their backbone structures. The side chains may include O-acetyl, phosphate, sialic acid, and other moieties. Those moieties represent the immunodominant epitopes and the most functional ones. The clinically significant Staphylococcus aureus type 5 CP (CP 5) and type 8 CP (CP 8) are comprised of a trisaccharide repeat unit with one O-acetyl group attached to each repeat unit. The immunogenicity of these CP and the functionality of antibodies to the backbone and the O-acetyl moieties were investigated. Immunization with the native CP conjugates (CP with 75% O-acetylation) elicited a high proportion of antibodies directed against the O-acetyl moiety. Nonetheless, all of the vaccinees produced antibodies to the backbone moieties as well. Conjugate vaccines made of de-O acetylated CP elicited backbone antibodies only. Antibodies to both backbone and O-acetyl groups were found to be opsonic against S. aureus strains which varied in their O-acetyl content. Absorption studies with O-acetylated and de-O acetylated CP showed that (i) native CP conjugates generated antibodies to both backbone and O-acetyl groups and (ii) O-acetylated isolates were opsonized by both populations of antibodies while the non-O-acetylated strains were predominantly opsonized by the backbone antibodies. These results suggest that S. aureus CP conjugate vaccines elicit multiple populations of antibodies with diverse specificities. Moreover, the antibodies of different specificities (backbone or O-acetyl) are all functional and efficient against the variations in bacterial CP that may occur among clinically significant S. aureus pathogenic isolates. PMID- 9746553 TI - Evolution of host adaptation in Salmonella enterica. PMID- 9746555 TI - Streptococcus pyogenes serotype M1 encodes multiple pathways for entry into human epithelial cells. AB - The ability of a serotype M1 strain of Streptococcus pyogenes to efficiently invade A549 human lung epithelial cells was previously shown to be dependent on bacterial exposure to human or bovine serum proteins or synthetic peptides containing the sequence RGD. In this study, stimulation by invasion agonists was determined to be dependent on expression of the streptococcal cell surface protein, M1. Fetal bovine serum (FBS), fibronectin (Fn), the extracellular matrix protein laminin (Lm), and RGD-containing peptides were tested for their abilities to promote epithelial cell invasion and adherence by isogenic M1(+) and M1(-) strains of S. pyogenes. In the absence of an agonist, invasion and adherence were comparable for the two bacterial strains. FBS, Fn, and Lm stimulated invasion of the M1(+) strain as much as 70-fold but failed to significantly affect invasion by the M1(-) mutant. Adherence of the wild-type strain was stimulated by these same agonists. Epithelial cell adherence by the M1(-) strain, however, was unaffected by the presence of Fn or Lm. Several RGD-containing peptides were found to promote invasion independently of M1 expression. Binding of 125I-Fn was reduced 88% by the M1(-) mutation and Fn was found to bind purified M1 protein, suggesting that Fn mediates invasion by direct binding to M1. To determine if host integrins might be involved in internalization of streptococci, several anti integrin monoclonal antibodies (MAbs) were tested for their abilities to inhibit invasion. Antibody directed against integrin beta1 inhibited FBS-, Fn-, and Lm mediated invasion but did not abrogate RGD-peptide-stimulated invasion. MAb directed against the epithelial cell Fn receptor, integrin alpha5beta1, inhibited Fn and FBS-mediated invasion but did not specifically inhibit Lm-mediated invasion. These results indicate that S. pyogenes has evolved multiple mechanisms for invasion of eukaryotic cells, at least two of which involve interactions between M1 protein, host integrins, and integrin ligands. PMID- 9746556 TI - Surface-associated hsp60 chaperonin of Legionella pneumophila mediates invasion in a HeLa cell model. AB - HeLa cells have been previously used to demonstrate that virulent strains of Legionella pneumophila (but not salt-tolerant avirulent strains) efficiently invade nonphagocytic cells. Hsp60, a member of the GroEL family of chaperonins, is displayed on the surface of virulent L. pneumophila (R. A. Garduno et al., J. Bacteriol. 180:505-513, 1988). Because Hsp60 is largely involved in protein protein interactions, we investigated its role in adherence-invasion in the HeLa cell model. Hsp60-specific antibodies inhibited the adherence and invasiveness of two virulent L. pneumophila strains in a dose-dependent manner but had no effect on the association of their salt-tolerant avirulent derivatives with HeLa cells. A monospecific anti-OmpS (major outer membrane protein) serum inhibited the association of both virulent and avirulent strains of L. pneumophila to HeLa cells, suggesting that while both Hsp60 and OmpS may mediate bacterial association to HeLa cells, only virulent strains selectively displayed Hsp60 on their surfaces. Furthermore, the surface-associated Hsp60 of virulent bacterial cells was susceptible to the action of trypsin, which rendered the bacteria noninvasive. Additionally, pretreatment of HeLa cells with purified Hsp60 or precoating of the plastic surface where HeLa cells attached with Hsp60 reduced the adherence and invasiveness of the two virulent strains. Finally, recombinant Hsp60 covalently bound to latex beads promoted the early association of beads with HeLa cells by a factor of 20 over bovine serum albumin (BSA)-coated beads and competed with virulent strains for association with HeLa cells. Hsp60-coated beads were internalized in large numbers by HeLa cells and remained in tight endosomes that did not fuse with other vesicles, whereas internalized BSA-coated beads, for which endocytic trafficking is well established, resided in more loose or elongated endosomes. Mature intracellular forms of L. pneumophila, which were up to 100-fold more efficient than agar-grown bacteria at associating with HeLa cells, were enriched for Hsp60 on the bacterial surface, as determined by immunolocalization techniques. Collectively, these results establish a role for surface-exposed Hsp60 in invasion of HeLa cells by L. pneumophila. PMID- 9746558 TI - Invasion by Salmonella typhimurium induces increased expression of the LMP, MECL, and PA28 proteasome genes and changes in the peptide repertoire of HLA-B27. AB - We have analyzed proteasomal adaptation and associated changes in the B27-bound peptide repertoire in response to cellular invasion with Salmonella. The peptide repertoire of HLA-B27 complexes was analyzed by two different methods: (i) high pressure liquid chromatography (HPLC) profiles of newly synthesized peptides eluted from B27 following metabolic labeling with arginine and (ii) reactivities with two B27 monoclonal antibodies, Ye-2 and B27.M2, sensitive to peptide-induced conformational changes. LMP, MECL, and PA28 expression was analyzed by reverse transcription-PCR (RT-PCR) of mRNA and by Western blot analysis for LMP2. Invasion of HLA-B27-transfected HeLa cells by Salmonella typhimurium induced significant changes in the reactivities of HLA-B27 with these two antibodies, which was accompanied by significant quantitative and qualitative changes in the HPLC profile of peptides eluted from HLA-B27. We also observed increases in the RT-PCR values for the LMP2, LMP7, and MECL proteasome subunit genes, as well as the proteasomal activator PA28alpha and -beta genes, and increased expression of the LMP2 protein by Western blotting. Upregulation of LMP2, but not LMP7, gene expression showed a close correlation with the changes in antibody reactivities observed upon bacterial invasion. We observed similar changes in reactivity with the Ye-2 or the B27.M2 antibody of lymphoblastoid cells upon gamma interferon treatment, which significantly correlated with the increased RT-PCR values for the LMP2 gene. This was accompanied by consistent HPLC profile changes for eluted peptides. Thus, Salmonella invasion leads to serologically recognizable changes in the B27-bound peptide repertoire, which may include peptides of host origin potentially through modulation of proteasome LMP2 subunit expression and, as a consequence, proteasomal activities. PMID- 9746557 TI - DNA sequencing and analysis of the low-Ca2+-response plasmid pCD1 of Yersinia pestis KIM5. AB - The low-Ca2+-response (LCR) plasmid pCD1 of the plague agent Yersinia pestis KIM5 was sequenced and analyzed for its genetic structure. pCD1 (70,509 bp) has an IncFIIA-like replicon and a SopABC-like partition region. We have assigned 60 apparently intact open reading frames (ORFs) that are not contained within transposable elements. Of these, 47 are proven or possible members of the LCR, a major virulence property of human-pathogenic Yersinia spp., that had been identified previously in one or more of Y. pestis or the enteropathogenic yersiniae Yersinia enterocolitica and Yersinia pseudotuberculosis. Of these 47 LCR-related ORFs, 35 constitute a continuous LCR cluster. The other LCR-related ORFs are interspersed among three intact insertion sequence (IS) elements (IS100 and two new IS elements, IS1616 and IS1617) and numerous defective or partial transposable elements. Regional variations in percent GC content and among ORFs encoding effector proteins of the LCR are additional evidence of a complex history for this plasmid. Our analysis suggested the possible addition of a new Syc- and Yop-encoding operon to the LCR-related pCD1 genes and gave no support for the existence of YopL. YadA likely is not expressed, as was the case for Y. pestis EV76, and the gene for the lipoprotein YlpA found in Y. enterocolitica likely is a pseudogene in Y. pestis. The yopM gene is longer than previously thought (by a sequence encoding two leucine-rich repeats), the ORF upstream of ypkA-yopJ is discussed as a potential Syc gene, and a previously undescribed ORF downstream of yopE was identified as being potentially significant. Eight other ORFs not associated with IS elements were identified and deserve future investigation into their functions. PMID- 9746559 TI - Binding properties of Streptococcus gordonii SspA and SspB (antigen I/II family) polypeptides expressed on the cell surface of Lactococcus lactis MG1363. AB - The oral bacterium Streptococcus gordonii expresses two cell wall-associated polypeptides, designated SspA (1,542 amino acid residues) and SspB (1,462 amino acid residues), that have 70% sequence identity. These polypeptides are members of the antigen I/II family of oral streptococcal adhesins and mediate the binding of streptococci to salivary glycoproteins, collagen, and other oral microorganisms such as Actinomyces naeslundii. To determine if SspA and SspB have differential binding properties, the coding sequences of the sspA and sspB genes were cloned into expression plasmid vector pTREX1-usp45LS to generate pTREX1-sspA and pTREX1-sspB, respectively, and the Ssp polypeptides were displayed on the cell surface of Lactococcus lactis MG1363. Lactococcal cells expressing similar levels of surface SspA or SspB polypeptide were then compared for their abilities to adhere to a range of antigen I/II polypeptide substrates. More than twice as many L. lactis cells expressing SspA bound to immobilized salivary agglutinin glycoprotein (SAG) as did L. lactis cells expressing SspB. In contrast, lactococci expressing SspB adhered twice as well as lactococci producing SspA to collagen type I and to Candida albicans. The binding of A. naeslundii to lactococci was only weakly enhanced by surface expression of Ssp polypeptides. L. lactis(pTREX1-sspB) cells bound in greater numbers to SAG than did Enterococcus faecalis JH2-2 cells expressing SspB from pAM401EB-5. The results suggest that SspA and SspB have markedly different binding affinities for their oral substrates and thus may function to promote site diversity in colonization by S. gordonii. PMID- 9746561 TI - Cytotoxic T-lymphocyte-mediated lysis of Toxoplasma gondii-infected target cells does not lead to death of intracellular parasites. AB - CD8(+) T cells play a crucial role in the control of infection with intracellular microbes. The mechanisms underlying the CD8(+) T-cell-mediated clearance of the intracellular pathogen Toxoplasma gondii are, however, not completely understood. The effect of CD8(+) cytotoxic T-lymphocyte (CTL)-mediated lysis of host cells on the viability of intracellular T. gondii was investigated. Quantitative competitive PCR of the gene encoding T. gondii major surface antigen (SAG-1) was combined with treatment of the parasites with DNase, which removed the DNA template of nonviable parasites. The induction by CD8(+) CTLs of apoptosis in cells infected with T. gondii did not result in the reduction of live parasites, indicating that intracellular T. gondii remains alive after lysis of host cells by CTLs. PMID- 9746560 TI - A second two-component regulatory system of Bordetella bronchiseptica required for bacterial resistance to oxidative stress, production of acid phosphatase, and in vivo persistence. AB - Random minitransposon mutagenesis was used to identify genes involved in the survival of Bordetella bronchiseptica within eukaryotic cells. One of the mutants which exhibited a reduced ability to survive intracellularly harbored a minitransposon insertion in a locus (ris) which displays a high degree of homology to two-component regulatory systems. This system exhibited less than 25% amino acid sequence homology to the only other two-component regulatory system described in Bordetella spp., the bvg locus. A risA'-'lacZ translational fusion was constructed and integrated into the chromosome of B. bronchiseptica. Determination of beta-galactosidase activity under different environmental conditions suggested that ris is regulated independently of bvg and is optimally expressed at 37 degrees C, in the absence of Mg2+, and when bacteria are in the intracellular niche. This novel regulatory locus, present in all Bordetella spp., is required for the expression of acid phosphatase by B. bronchiseptica. Although catalase and superoxide dismutase production were unaffected, the ris mutant was more sensitive to oxidative stress than the wild-type strain. Complementation of bvg-positive and bvg-negative ris mutants with the intact ris operon incorporated as a single copy into the chromosome resulted in the reestablishment of the ability of the bacterium to produce acid phosphatase and to resist oxidative stress. Mouse colonization studies demonstrated that the ris mutant is cleared by the host much earlier than the wild-type strain, suggesting that ris-regulated products play a significant role in natural infections. The identification of a second two-component system in B. bronchiseptica highlights the complexity of the regulatory network needed for organisms with a life cycle requiring adaptation to both the external environment and a mammalian host. PMID- 9746562 TI - Genetic divergence and evolutionary instability in ospE-related members of the upstream homology box gene family in Borrelia burgdorferi sensu lato complex isolates. AB - A series of related genes that are flanked at their 5' ends by a conserved upstream sequence element called the upstream homology box (UHB) have been identified in Borrelia burgdorferi. These genes have been referred to as the UHB or erp gene family. We previously demonstrated that among a limited number of B. burgdorferi isolates, the UHB gene family is variable in composition and organization. Prior to this report the UHB gene family in other species of the B. burgdorferi sensu lato complex had not been studied, and if this family is important in the pathogenesis or biology of the Lyme disease spirochetes, then a wide distribution among species and isolates of the B. burgdorferi sensu lato complex would be expected. To assess this, we screened for the UHB element by Southern hybridization and determined its restriction fragment length polymorphism (RFLP) patterns. The UHB element was found to be carried by all B. burgdorferi sensu lato complex species tested (B. burgdorferi, B. garinii, B. afzelii, B. japonica, B. valaisiana sp. nov., and B. andersonii), but the RFLP patterns varied widely at both the inter- and intraspecies levels. Variation in both the number and size of the hybridizing restriction fragments was evident. PCR analyses also revealed the presence of polymorphic, ospE-related alleles in many isolates. Sequence analyses identified the molecular basis of the polymorphisms as being primarily insertions and deletions. Sequence variation and the insertions and deletions were found to be clustered in two distinct domains (variable domains 1 and 2). In many isolates variable domain 1 is flanked by direct repeat elements, some as long as 38 bp. Computer analyses of the deduced amino acid sequences encoded within variable domain 1 predict them to be hydrophilic, surface exposed, and antigenic. The analyses conducted here suggest that the UHB gene family, as evidenced by the variable UHB RFLP patterns, is not evolutionarily stable and that the polymorphic ospE alleles are derived from a common ancestral gene which has been modified through mutation or recombination events. The characterization of ospE-related genes of the UHB gene family among B. burgdorferi sensu lato species will prove important in attempts to construct a model for UHB gene family organization and in deciphering the role of the UHB gene family in the biology and pathogenesis of the Lyme disease spirochetes. PMID- 9746563 TI - Induction of adrenomedullin mRNA and protein by lipopolysaccharide and paclitaxel (Taxol) in murine macrophages. AB - Lipopolysaccharide (LPS), a potent inflammatory stimulus derived from the outer membrane of gram-negative bacteria, has been implicated in septic shock. Plasma levels of adrenomedullin (AM), a potent vasorelaxant, are increased in septic shock and possibly contribute to the characteristic hypotension. As macrophages play a central role in the host response to LPS, we studied AM production by LPS stimulated macrophages. When peritoneal exudate macrophages from C3H/OuJ mice were treated with protein-free LPS (100 ng/ml) or the LPS mimetic paclitaxel (Taxol; 35 microM), an approximately 10-fold increase in steady-state AM mRNA levels was observed, which peaked between 2 and 4 h. A three- to fourfold maximum increase in the levels of immunoreactive AM protein was detected after 6 to 8 h of stimulation. While LPS-hyporesponsive C3H/HeJ macrophages failed to respond to protein-free LPS with an increase in steady-state AM mRNA levels, increased levels were observed after stimulation of these cells with a protein-rich (butanol-extracted) LPS preparation. In addition, increased AM mRNA was observed following treatment of either C3H/OuJ or C3H/HeJ macrophages with soluble Toxoplasma gondii tachyzoite antigen or the synthetic flavone analog 5, 6 dimethylxanthenone-4-acetic acid. Gamma interferon also stimulated C3H/OuJ macrophages to express increased AM mRNA levels yet was inhibitory in the presence of LPS or paclitaxel. In vivo, mice challenged intraperitoneally with 25 microg of LPS exhibited increased AM mRNA levels in the lungs, liver, and spleen; the greatest increase (>50-fold) was observed in the liver and lungs. Thus, AM is produced, by murine macrophages, and furthermore, LPS induces AM mRNA in vivo in a number of tissues. These data support a possible role for AM in the pathophysiology of sepsis and septic shock. PMID- 9746564 TI - Filament tip-associated antigens involved in adherence to and invasion of murine pulmonary epithelial cells in vivo and HeLa cells in vitro by Nocardia asteroides. AB - The interactions of Nocardia asteroides GUH-2 with pulmonary epithelial cells of C57BL/6 mice and with HeLa cells were studied. Electron microscopy demonstrated that only the tips of log-phase cells penetrated pulmonary epithelial cells following intranasal administration, and nocardiae were recovered from the brain. Coccobacillary cells neither invaded nor disseminated. Serum from immunized mice (IMS) decreased attachment to and penetration of pulmonary epithelial cell surfaces by log-phase GUH-2 and inhibited spread to the brain. IMS was adsorbed against stationary-phase cells. Western immunoblots suggested that this adsorbed IMS was reactive primarily with 43- and 62-kDa proteins. Immunofluorescence showed that adsorbed IMS preferentially labeled the tips of log-phase GUH-2 cells. Since this IMS was reactive to culture filtrate antigens, several of these proteins were cut from gels, and mice were immunized. Sera against 62-, 55-, 43-, 36-, 31-, and 25-kDa antigens were obtained. The antisera against the 43- and 36 kDa proteins labeled the filament tips of GUH-2 cells. Only the antiserum against the 43-kDa antigen increased pulmonary clearance, inhibited apical attachment to and penetration of pulmonary epithelial cells, and prevented spread to the brain. An in vitro model with HeLa cells demonstrated that the tips of log-phase cells of GUH-2 adhered to and penetrated the surface of HeLa cells. Invasion assays with amikacin treatment demonstrated that nocardiae were internalized. Adsorbed IMS blocked attachment to and invasion of these cells. These data suggested that a filament tip-associated 43-kDa protein was involved in attachment to and invasion of pulmonary epithelial cells and HeLa cells by N. asteroides GUH-2. PMID- 9746566 TI - Anthrax toxin as a molecular tool for stimulation of cytotoxic T lymphocytes: disulfide-linked epitopes, multiple injections, and role of CD4(+) cells. AB - We have previously demonstrated that anthrax toxin-derived proteins, protective antigen (PA) and the amino-terminal portion of lethal factor (LFn), can be used in combination to deliver heterologous molecules to the cytosol of mammalian cells. In this study we examined the ability of an LFn-peptide disulfide-linked heterodimer to prime cytotoxic T lymphocytes (CTL) in the presence of PA. A mutant of LFn that contains a carboxy-terminal reactive cysteine was generated. This form of LFn could be oxidized with a synthetic cysteine containing peptide to form a heterodimer of the protein and peptide. Mice injected with the heterodimer plus PA mounted a peptide-specific CTL response, indicating that this molecule functioned similarly to the genetically fused forms used previously. We also report the results of an analysis of two aspects of this system important for the development of experimental vaccines. First, CD4 knockout mice were unable to generate a CTL response when treated with PA plus an LFn-epitope fusion protein, suggesting that CD4(+) helper responses are essential for stimulating specific CTL with the PA-LFn system. Second, we now show that primary injection with this system does not generate any detectable antibody response to the vaccine components and that prior immunization has no effect on priming a CTL response to an unrelated epitope upon subsequent injection. PMID- 9746565 TI - The cysteine-cysteine family of chemokines RANTES, MIP-1alpha, and MIP-1beta induce trypanocidal activity in human macrophages via nitric oxide. AB - This paper describes a new role for the cysteine-cysteine (CC) chemokines RANTES, MIP-1alpha, and MIP-1beta on human macrophage function, which is the induction of nitric oxide (NO)-mediated trypanocidal activity. In a previous report, we showed that RANTES, MIP-1alpha and MIP-1beta enhance Trypanosoma cruzi uptake and promote parasite killing by human macrophages (M. F. Lima, Y. Zhang, and F. Villalta, Cell. Mol. Biol. 43:1067-1076, 1997). Here we study the mechanism by which RANTES, MIP-1alpha, and MIP-1beta activate human macrophages obtained from healthy individuals to kill T. cruzi. Treatment of human macrophages with different concentrations of RANTES, MIP-1alpha, and MIP-1beta enhances T. cruzi trypomastigote phagocytosis in a dose peak response. The optimal response induced by the three CC chemokines is attained at 500 ng/ml. The macrophage trypanocidal activity induced by CC chemokines can be completely inhibited by L-N-monomethyl arginine (L-NMMA), a specific inhibitor of the L-arginine:NO pathway, but not by its D-enantiomer. Culture supernatants of chemokine-treated human macrophages contain increased NO2- levels, and NO2- production is also specifically inhibited by L-NMMA. The amount of NO2- induced by these chemokines in human macrophages is comparable to the amount of NO2- induced by gamma interferon. The killing of trypomastigotes by NO in cell-free medium is blocked by an NO antagonist or a NO scavenger. This data supports the hypothesis that the CC chemokines RANTES, MIP 1alpha, and MIP-1beta activate human macrophages to kill T. cruzi via NO, which is an effective trypanocidal mechanism. PMID- 9746567 TI - Identification of two Shigella flexneri chromosomal loci involved in intercellular spreading. AB - The ability of Shigella flexneri to multiply within colonic epithelial cells and spread to adjacent cells is essential for production of dysentery. Two S. flexneri chromosomal loci that are required for these processes were identified by screening a pool of TnphoA insertion mutants. These mutants were able to invade cultured epithelial cells but could not form wild-type plaques. Analysis of the nucleotide sequence indicated that the sites of TnphoA insertion were within two different regions that are almost identical to Escherichia coli K-12 chromosomal sequences of unknown functions. One region is located at 70 min on the E. coli chromosome, upstream of murZ, while the other is at 28 min, downstream of tonB. The mutant with the insertion at 70 min was named vpsC because it showed an altered pattern of virulence protein secretion. The vpsC mutant formed pinpoint-sized plaques, was defective in recovery from infected tissue culture cells, and was sensitive to lysis by the detergent sodium dodecyl sulfate. Recombinant plasmids carrying the S. flexneri vpsA, -B, and -C genes complemented all of the phenotypes of the vpsC mutant. A mutation in vpsA resulted in the same phenotype as the vpsC mutation, suggesting that these two genes are part of a virulence operon in S. flexneri. The mutant with the insertion at 28 min was interrupted in the same open reading frame as S. flexneri ispA. This ispA mutant could not form plaques and was defective in bacterial septation inside tissue culture cells. PMID- 9746568 TI - The ica locus of Staphylococcus epidermidis encodes production of the capsular polysaccharide/adhesin. AB - Clinical isolates of coagulase-negative staphylococci often elaborate a biofilm involved in adherence to medical devices and resistance to host defenses. The biofilm contains the capsular polysaccharide/adhesin (PS/A), which mediates cell adherence to biomaterials, and another antigen, termed polysaccharide intercellular adhesin (PIA), which is thought to mediate bacterial accumulation into cellular aggregates. PIA is a polymer of beta-1, 6-linked N-acetyl glucosamine residues with a molecular mass of <30, 000 kDa. We found that recombinant Staphylococcus carnosus and Staphylococcus aureus carrying a plasmid with genes of the ica locus, which was reported to encode the biosynthetic proteins for production of PIA, were also able to synthesize PS/A. PS/A and a chemically and immunologically identical polysaccharide isolated from S. carnosus carrying the ica genes on plasmid pCN27 were found to be high-molecular-mass (>250,000 kDa), acid-stable polymers of beta-1,6-linked glucosamine substituted on the amino group primarily with succinate, although some preparations also contained acetate. Moreover, all recombinant staphylococcal strains with the ica genes had the biologic properties previously attributed to PS/A. ica-positive strains readily formed an in vitro biofilm on plastic, adhered 3- to 10-fold more to catheters during a 30-min assay compared with control strains carrying only the cloning vector, adsorbed out antibodies to PS/A from immune serum, and elaborated a capsule visualized by immunoelectron microscopy with antisera to PS/A. These properties were also seen with PS/A-producing strains of Staphylococcus epidermidis, but not with transposon mutants lacking PS/A. An antiserum raised to PIA contained high-titer antibody to PS/A that was readily adsorbed out by PS/A-positive strains of S. epidermidis and recombinant strains of staphylococci carrying the ica genes. We conclude that the ica locus encodes production of PS/A and that the properties of S. epidermidis associated with initial bacterial adherence, biofilm formation, and intercellular adhesion can be correlated with elaboration of PS/A. PMID- 9746569 TI - The number of direct repeats in hagA is variable among Porphyromonas gingivalis strains. AB - The coding sequence for the surface protein hemagglutinin A (HagA) of Porphyromonas gingivalis 381 has previously been shown to contain four direct 1.35-kb repeats, designated repHA. This study was performed to determine if the number of repHA units in hagA is consistently 4 or if allelic polymorphism exists among strains and/or upon multiple passage of P. gingivalis. To this end, primers which were homologous to the regions directly 5' and 3' of the repeat domain in hagA were synthesized. PCR conditions which allowed amplification of the 8.4-kb repeat region between the primers in P. gingivalis 381 were established. Genomic DNA templates from 13 other P. gingivalis strains and 9 fresh clinical isolates from patients were analyzed under the same conditions as used above. Analysis of these PCR products demonstrated that the strains tested had different numbers (two to four) of repHA units in the respective hagA genes. The PCR products of 8.4, 7.0, and 5.7 kb represent four, three, and two repeats, respectively. One strain from each group (381, four repeats; W83, three repeats; and AJW4, two repeats) was also tested to determine if the number of repeats remained invariant upon passaging onto solid medium. No variability in the number of repeats in hagA within a strain was detected after 18 passages. P. gingivalis 381 was chosen for further testing in a mouse abscess model to determine if conditions of in vivo growth would select for deletions or duplications of the repeated sequences. Five days after infection, no change in the number of repeats was detected in cells recovered from either nonimmunized or preimmunized mice. This data indicates an interstrain variability of the number of repeat units and hence a size variability of the HagA protein of P. gingivalis, but unlike some surface antigens of other pathogenic species, the number of repeats remains relatively stable given the conditions of growth tested here. PMID- 9746570 TI - Characterization of outer membrane protein OmpU of Vibrio cholerae O1. AB - The outer membrane protein OmpU of Vibrio cholerae O1 strain 86B3 was characterized with reference to colonization of the intestine by the organism. The purified OmpU exhibited a pI of 3.6. Upon sodium dodecyl sulfate polyacrylamide gel electrophoresis, it migrated to 38, 32, and 110 kDa when the sample was heated at 100 degrees C for 2 min, 50 degrees C for 15 min, and room temperature for 30 min, respectively. The purified OmpU was not hemagglutinative. Anti-OmpU serum did not agglutinate strain 86B3 or other V. cholerae organisms. OmpU adhered to the brush border of the rabbit small intestine; adhesion of the organisms to the intestine treated in advance with OmpU was not inhibited. Treating the organisms in advance with anti-OmpU Fab did not inhibit adhesion to the intestine. These results obtained in vitro suggest that OmpU is not involved in the adhesion of V. cholerae to the intestinal epithelium. PMID- 9746571 TI - Role of Fusobacterium nucleatum and coaggregation in anaerobe survival in planktonic and biofilm oral microbial communities during aeration. AB - Coaggregation is a well-characterized phenomenon by which specific pairs of oral bacteria interact physically. The aim of this study was to examine the patterns of coaggregation between obligately anaerobic and oxygen-tolerant species that coexist in a model oral microbial community. Obligate anaerobes other than Fusobacterium nucleatum coaggregated only poorly with oxygen-tolerant species. In contrast, F. nucleatum was able to coaggregate not only with both oxygen-tolerant and other obligately anaerobic species but also with otherwise-noncoaggregating obligate anaerobe-oxygen-tolerant species pairs. The effects of the presence or absence of F. nucleatum on anaerobe survival in both the biofilm and planktonic phases of a complex community of oral bacteria grown in an aerated (gas phase, 200 ml of 5% CO2 in air x min-1) chemostat system were then investigated. In the presence of F. nucleatum, anaerobes persisted in high numbers (>10(7) x ml-1 in the planktonic phase and >10(7) x cm-2 in 4-day biofilms). In an equivalent culture in the absence of F. nucleatum, the numbers of black-pigmented anaerobes (Porphyromonas gingivalis and Prevotella nigrescens) were significantly reduced (P /=0.5 microg/mouse. Serum neutralization and ELISA titers were also determined. Tellingly, 82 of 83 mice with antibody titers of >/=1, 600, as measured by ELISA, survived, but only 6 of 42 mice with titers of 10(4) parasites), while a mutant VEG strain (MS-J) that is unable to growth shift caused 100% mortality in mice inoculated with 10 parasites. Parasites recovered from gamma interferon knockout mice inoculated with emergent tachyzoites grew at a slow rate and expressed BAG1, confirming that the replication switch occurs in animals and in the absence of a protective immune response. PMID- 9746588 TI - Mannan-specific immunoglobulin G antibodies in normal human serum accelerate binding of C3 to Candida albicans via the alternative complement pathway. AB - Candida albicans activates the classical and alternative complement pathways, leading to deposition of opsonic complement fragments on the cell surface. Our previous studies found that antimannan immunoglobulin G (IgG) in normal human serum (NHS) allows C. albicans to initiate the classical pathway. The purpose of this study was to determine whether antimannan IgG also plays a role in initiation of the alternative pathway. Pooled NHS was rendered free of classical pathway activity by chelation of serum Ca2+ with EGTA alone or in combination with immunoaffinity removal of antimannan antibodies. Kinetic analysis revealed a 6-min lag in detection of C3 binding to C. albicans incubated in EGTA-chelated NHS, compared to a 12-min lag in NHS that was both EGTA chelated and mannan absorbed. The 12-min lag was shortened to 6 min by addition of affinity-purified antimannan IgG. The accelerating effect of antimannan IgG on alternative pathway initiation was dose dependent and was reproduced in a complement binding reaction consisting of six purified proteins of the alternative pathway. Both Fab and F(ab')2 fragments of antimannan IgG facilitated alternative pathway initiation in a manner similar to that observed with intact antibody. Immunofluorescence analysis showed that addition of antimannan IgG to EGTA-chelated and mannan absorbed serum promoted an early deposition of C3 molecules on the yeast cells but had little or no effect on distribution of the cellular sites for C3 activation. Thus, antimannan IgG antibodies play an important regulatory role in interactions between the host complement system and C. albicans. PMID- 9746589 TI - Characterization of the hemorrhagic reaction caused by Vibrio vulnificus metalloprotease, a member of the thermolysin family. AB - Vibrio vulnificus is an opportunistic human pathogen causing wound infections and septicemia, characterized by hemorrhagic and edematous damage to the skin. This human pathogen secretes a metalloprotease (V. vulnificus protease [VVP]) as an important virulence determinant. When several bacterial metalloproteases including VVP were injected intradermally into dorsal skin, VVP showed the greatest hemorrhagic activity. The level of the in vivo hemorrhagic activity of the bacterial metalloproteases was significantly correlated with that of the in vitro proteolytic activity for the reconstituted basement membrane gel. Of two major basement membrane components (laminin and type IV collagen), only type IV collagen was easily digested by VVP. Additionally, the immunoglobulin G antibody against type IV collagen, but not against laminin, showed sufficient protection against the hemorrhagic reaction caused by VVP. Capillary vessels are known to be stabilized by binding of the basal surface of vascular endothelial cells to the basement membrane. Therefore, specific degradation of type IV collagen may cause destruction of the basement membrane, breakdown of capillary vessels, and leakage of blood components including erythrocytes. PMID- 9746590 TI - Experimental infection of Mongolian gerbils with wild-type and mutant Helicobacter pylori strains. AB - Experimental Helicobacter pylori infection was studied in Mongolian gerbils with fresh human isolates that carry or do not carry cagA (cagA-positive or cagA negative, respectively), multiply passaged laboratory strains, wild-type strain G1.1, or isogenic ureA, cagA, or vacA mutants of G1.1. Animals were sacrificed 1 to 32 weeks after challenge, the stomach was removed from each animal for quantitative culture, urease test, and histologic testing, and blood was collected for antibody determinations. No colonization occurred after >/=20 in vitro passages of wild-type strain G1.1 or with the ureA mutant of G1.1. In contrast, infection occurred in animals challenged with wild-type G1.1 (99 of 101 animals) or the cagA (25 of 25) or vacA (25 of 29) mutant of G1.1. Infection with G1.1 persisted for at least 8 months. All 15 animals challenged with any of three fresh human cagA-positive isolates became infected, in contrast to only 6 (23%) of 26 animals challenged with one of four fresh human cagA-negative isolates (P < 0.001). Similar to infection in humans, H. pylori colonization of gerbils induced gastric inflammation and a systemic antibody response to H. pylori antigens. These data confirm the utility of gerbils as an animal model of H. pylori infection and indicate the importance of bacterial strain characteristics for successful infection. PMID- 9746591 TI - Murine dendritic cells pulsed in vitro with Toxoplasma gondii antigens induce protective immunity in vivo. AB - The activation of a predominant T-helper-cell subset plays a critical role in disease resolution. In the case of Toxoplasma gondii, the available evidence indicates that CD4(+) protective cells belong to the Th1 subset. The aim of this study was to determine whether T. gondii antigens (in T. gondii sonicate [TSo]) presented by splenic dendritic cells (DC) were able to induce a specific immune response in vivo and to protect CBA/J mice orally challenged with T. gondii cysts. CBA/J mice immunized with TSo-pulsed DC exhibited significantly fewer cysts in their brains after oral infection with T. gondii 76K than control mice did. Protected mice developed a strong humoral response in vivo, with especially high levels of anti-TSo immunoglobulin G2a antibodies in serum. T. gondii antigens such as SAG1 (surface protein), SAG2 (surface protein), MIC1 (microneme protein), ROP2 through ROP4 (rhoptry proteins), and MIC2 (microneme protein) were recognized predominantly. Furthermore, DC loaded with TSo, which synthesized high levels of interleukin-12 (IL-12), triggered a strong cellular response in vivo, as assessed by the proliferation of lymph node cells in response to TSo restimulation in vitro. Cellular proliferation was associated with gamma interferon and IL-2 production. Taken together, these results indicate that immunization of CBA/J mice with TSo-pulsed DC can induce both humoral and Th1 like cellular immune responses and affords partial resistance against the establishment of chronic toxoplasmosis. PMID- 9746592 TI - Borrelia burgdorferi and interleukin-1 promote the transendothelial migration of monocytes in vitro by different mechanisms. AB - A prominent feature of Lyme disease is the perivascular accumulation of mononuclear leukocytes. Incubation of human umbilical vein endothelial cells (HUVEC) cultured on amniotic tissue with either interleukin-1 (IL-1) or Borrelia burgdorferi, the spirochetal agent of Lyme disease, increased the rate at which human monocytes migrated across the endothelial monolayers. Very late antigen 4 (VLA-4) and CD11/CD18 integrins mediated migration of monocytes across HUVEC exposed to either B. burgdorferi or IL-1 in similar manners. Neutralizing antibodies to the chemokine monocyte chemoattractant protein 1 (MCP-1) inhibited the migration of monocytes across unstimulated, IL-1-treated, or B. burgdorferi stimulated HUVEC by 91% +/- 3%, 65% +/- 2%, or 25% +/- 22%, respectively. Stimulation of HUVEC with B. burgdorferi also promoted a 6-fold +/- 2-fold increase in the migration of human CD4(+) T lymphocytes. Although MCP-1 played only a limited role in the migration of monocytes across B. burgdorferi-treated HUVEC, migration of CD4(+) T lymphocytes across HUVEC exposed to spirochetes was highly dependent on this chemokine. The anti-inflammatory cytokine IL-10 reduced both migration of monocytes and endothelial production of MCP-1 in response to B. burgdorferi by approximately 50%, yet IL-10 inhibited neither migration nor secretion of MCP-1 when HUVEC were stimulated with IL-1. Our results suggest that activation of endothelium by B. burgdorferi may contribute to formation of the chronic inflammatory infiltrates associated with Lyme disease. The transendothelial migration of monocytes that is induced by B. burgdorferi is significantly less dependent on MCP-1 than is migration induced by IL-1. Selective inhibition by IL-10 further indicates that B. burgdorferi and IL-1 employ distinct mechanisms to activate endothelial cells. PMID- 9746593 TI - Antigenic analysis of Bordetella pertussis filamentous hemagglutinin with phage display libraries and rabbit anti-filamentous hemagglutinin polyclonal antibodies. AB - Although substantial advancements have been made in the development of efficacious acellular vaccines against Bordetella pertussis, continued progress requires better understanding of the antigenic makeup of B. pertussis virulence factors, including filamentous hemagglutinin (FHA). To identify antigenic regions of FHA, phage display libraries constructed by using random fragments of the 10 kbp EcoRI fragment of B. pertussis fhaB were affinity selected with rabbit anti FHA polyclonal antibodies. Characterization of antibody-reactive clones displaying FHA-derived peptides identified 14 antigenic regions, each containing one or more epitopes. A number of clones mapped within regions containing known or putative FHA adhesin domains and may be relevant for the generation of protective antibodies. The immunogenic potential of the phage-displayed peptides was assessed indirectly by comparing their recognition by antibodies elicited by sodium dodecyl sulfate (SDS)-denatured and native FHA and by measuring the inhibition of this recognition by purified FHA. FHA residues 1929 to 2019 may contain the most dominant linear epitope of FHA. Clones mapping to this region accounted for ca. 20% of clones recovered from the initial library selection and screening procedures. They are strongly recognized by sera against both SDS denatured and native FHA, and this recognition is readily inhibited by purified FHA. Given also that this region includes a factor X homolog (J. Sandros and E. Tuomanen, Trends Microbiol. 1:192-196, 1993) and that the single FHA epitope (residues 2001 to 2015) was unequivocally defined in a comparable study by E. Leininger et al. (J. Infect. Dis. 175:1423-1431, 1997), peptides derived from residues of 1929 to 2019 of FHA are strong candidates for future protection studies. PMID- 9746594 TI - Surface structure, hydrophobicity, phagocytosis, and adherence to matrix proteins of Bacillus cereus cells with and without the crystalline surface protein layer. AB - Nonopsonic phagocytosis of Bacillus cereus by human polymorphonuclear leukocytes (PMNs) with particular attention to bacterial surface properties and structure was studied. Two reference strains (ATCC 14579(T) and ATCC 4342) and two clinical isolates (OH599 and OH600) from periodontal and endodontic infections were assessed for adherence to matrix proteins, such as type I collagen, fibronectin, laminin, and fibrinogen. One-day-old cultures of strains OH599 and OH600 were readily ingested by PMNs in the absence of opsonins, while cells from 6-day-old cultures were resistant. Both young and old cultures of the reference strains of B. cereus were resistant to PMN ingestion. Preincubation of PMNs with the phagocytosis-resistant strains of B. cereus did not affect the phagocytosis of the sensitive strain. Negatively stained cells of OH599 and OH600 studied by electron microscopy had a crystalline protein layer on the cell surface. In thin sectioned cells of older cultures (3 to 6 days old), the S-layer was observed to peel off from the cells. No S-layer was detected on the reference strains. Extraction of cells with detergent followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed a major 97-kDa protein from the strains OH599 and OH600 but only a weak 97-kDa band from the reference strain ATCC 4342. One-day-old cultures of the clinical strains (hydrophobicity, 5.9 to 6.0%) showed strong binding to type I collagen, laminin, and fibronectin. In contrast, reference strains (hydrophobicity, -1.0 to 4.2%) as well as 6-day-old cultures of clinical strains (hydrophobicity, 19.0 to 53.0%) bound in only low numbers to the proteins. Gold-labelled biotinylated fibronectin was localized on the S-layer on the cell surface as well as on fragments of S-layer peeling off the cells of a 6-day-old culture of B. cereus OH599. Lactose, fibronectin, laminin, and antibodies against the S-protein reduced binding to laminin but not to fibronectin. Heating the cells at 84 degreesC totally abolished binding to both proteins. Benzamidine, a noncompetitive serine protease inhibitor, strongly inhibited binding to fibronectin whereas binding to laminin was increased. Overall, the results indicate that changes in the surface structure, evidently involving the S-layer, during growth of the clinical strains of B. cereus cause a shift from susceptibility to PMN ingestion and strong binding to matrix and basement membrane proteins. Furthermore, it seems that binding to laminin is mediated by the S-protein while binding to fibronectin is dependent on active protease evidently attached to the S-layer. PMID- 9746596 TI - Intestinal secretory factor released by macrophages stimulated with Clostridium difficile toxin A: role of interleukin 1beta. AB - Clostridium difficile toxin A is associated with enterocolitis in animals and humans. However, the mechanisms of its secretory and damaging effects are not totally understood. In this work, we examined the intestinal secretion of electrolytes and water caused by supernatants from macrophages stimulated with toxin A in rabbit ileal mucosa mounted in Ussing chambers. We also investigated the mechanism by which the intestinal secretory factor (ISF) is released from stimulated macrophages. Supernatants from macrophages stimulated with toxin A caused potent intestinal secretion (change in short-circuit current [DeltaIsc], 76 microA x cm-2; P < 0.01). The release of the ISF was pertussis toxin sensitive (reduction, 61%; P < 0.01) and was also reduced (P < 0.05) by a protein synthesis inhibitor (67%), protease inhibitors (57%), a phospholipase A2 inhibitor (54%), a cyclo-oxygenase inhibitor (62%), a dual cyclo- and lipoxygenase inhibitor (48%), a platelet-activating factor (PAF) receptor antagonist (55%), and tumor necrosis factor alpha (TNF-alpha) synthesis inhibitors (48%). However, this release was not inhibited by a lipo-oxygenase inhibitor. Monoclonal anti-interleukin 1beta (IL-1beta) but not anti-IL-1alpha antibody blocked (72%; P < 0.01) the secretory action of the ISF, as did recombinant human IL-1 receptor antagonist (80%; P < 0.01). High levels of IL-1beta (3,476 pg/ml) were detected by an enzyme-linked immunosorbent assay in the above supernatants. Furthermore, the addition of IL 1beta to the serosal side caused a potent secretory effect (DeltaIsc, 80 microA x cm-2; P < 0.01). These results show that macrophages stimulated with toxin A release an ISF capable of provoking intestinal secretion. The regulation of this factor is dependent upon the activation of the G protein. In addition, prostaglandins, PAF, and TNF-alpha are involved in the release of the ISF. We conclude that IL-1beta is probably the ISF released by macrophages in response to toxin A. PMID- 9746597 TI - Effects of orally administered epidermal growth factor on enteropathogenic Escherichia coli infection in rabbits. AB - The increased intestinal absorption induced by epidermal growth factor (EGF) is associated with diffuse lengthening of brush border microvilli. The aim of this study was to examine the in vivo effects of oral administration of EGF during infection with enteropathogenic Escherichia coli. New Zealand White rabbits (4 weeks old) received orogastric EGF daily starting 3 days prior to infection with enteropathogenic E. coli RDEC-1 and were compared with sham-treated infected animals and uninfected controls. Weight gain, food intake, fecal E. coli, and stool consistency were assessed daily. On day 10, segments of jejunum, ileum, proximal, and distal colon were assessed for gram-negative bacterial colonization, disaccharidase activities, and epithelial ultrastructure. Effects of EGF on E. coli RDEC-1 proliferation were studied in vitro. E. coli RDEC-1 caused diarrhea and reduced weight gain. Seven days postinfection, the small and large intestines were colonized with numerous bacteria, brush border microvilli were disrupted, and maltase and sucrase activities were significantly reduced in the jejunum. Daily treatment with EGF prevented the occurrence of diarrhea and reduction of weight gain. These effects were associated with significant inhibition of E. coli colonization in the small and large intestine, improved jejunal maltase and sucrase activities and reduced microvillous injury. EGF did not affect the proliferation of E. coli in vitro. The findings suggest that EGF protects the gastrointestinal tract against colonization by enteropathogenic E. coli. PMID- 9746595 TI - T-cell recognition of mycobacterial GroES peptides in Thai leprosy patients and contacts. AB - We report here the mapping of T-cell-stimulatory determinants of the GroES 10-kDa heat shock protein homologues from Mycobacterium leprae and Mycobacterium tuberculosis, which are known as major immunogens in mycobacterial infections. Peripheral blood mononuclear cells (PBMC) from treated tuberculoid leprosy or lepromatous leprosy patients and from healthy household or hospital staff contacts of the patients were cultured with 20 16-mer peptides covering the entire sequences of both M. leprae and M. tuberculosis GroES. The total number of recognized peptides was found to be the largest in family contacts, while responder frequencies to the individual tested peptides varied (5 to 80%) with specificity between the patient and contact groups. Proliferative responses to some peptides showed positive or negative associations of low statistical significance with DR and DQ alleles, though responses to most GroES peptides were genetically permissive. Notably, the sequence of the 25-40 peptide of M. leprae, but not that of M. tuberculosis, was more frequently stimulatory in tuberculoid leprosy patients than in either group of sensitized healthy contacts. This peptide bound to a number of HLA-DR molecules, of which HLA-DRB5*0101 had the strongest affinity. The epitope core binding to this allele was localized to the 29-to-37 sequence, and its key residue was localized to the M. leprae-specific glutamic acid at position 32. This epitope may be of interest for the development of a blood test- or skin test-based diagnostic reagent for tuberculoid leprosy, subject to further clinical evaluation in untreated patients. PMID- 9746598 TI - Cloning and molecular characterization of a cDNA clone coding for Trichomonas vaginalis alpha-actinin and intracellular localization of the protein. AB - We have identified and sequenced a cDNA clone coding for Trichomonas vaginalis alpha-actinin. Analysis of the obtained sequence revealed that the 2,857 nucleotide-long cDNA contained an open reading frame encoding 849 amino acids which showed consistent homology with alpha-actinins of different species. Such homology was particularly significant in regions which have been reported to represent the actin-binding and Ca2+-binding domains in other alpha-actinins. The deduced protein was also characterized by the presence of a divergent central region thought to play a role in its high immunogenicity. A study of protein localization performed by immunofluorescence revealed that the protein is diffusely distributed throughout the T. vaginalis cytoplasm when the cell is pear shaped. When parasites adhere and transform into the amoeboid morphology, the protein is located only in areas close to the cytoplasmic membrane and colocalizes with actin. Concomitantly with transformation into the amoeboid morphology, alpha-actinin mRNA expression is upregulated. PMID- 9746599 TI - Characterization of group B streptococcal invasion of human chorion and amnion epithelial cells In vitro. AB - Group B streptococci (GBS) have been cultured from the chorioamnionic membrane of pregnant women, usually in association with chorioamnionitis and premature labor (K. A. Boggess, D. H. Watts, S. L. Hillier, M. A. Krohn, T. J. Benedetti, and D. A. Eschenbach, Obstet. Gynecol. 87:779-784, 1996). Colonization and infection of placental membranes can be a prelude to neonatal GBS infections even in the presence of intact membranes (R. L. Naeye and E. C. Peters, Pediatrics 61:171 177, 1978), suggesting that GBS cause chorioamnionitis or establish amniotic fluid infections by partial or complete penetration of the placental membranes. We have isolated and grown cultures of primary chorion and amnion cells from human cesarean-section placentas. This has provided a biologically relevant model for investigating GBS adherence to and invasion of the two epithelial barriers of the placental membrane. GBS adhered to chorion cell monolayers to a high degree. Pretreatment of GBS with trypsin reduced adherence up to 10-fold, which suggested that the bacterial ligand(s) was a protein. GBS invaded chorion cells at a high rate in vitro, and invasion was dependent on cellular actin polymerization. GBS could be seen within intracellular vacuoles of chorion cells by transmission electron microscopy. We also demonstrated that GBS were capable of transcytosing through intact chorion cell monolayers without disruption of intracellular junctions. GBS also adhered to amnion cells; in contrast, however, these bacteria failed to invade amnion cells under a variety of assay conditions. GBS interactions with the chorion epithelial cell layer shown here correlate well with epidemiological and pathological studies of GBS chorioamnionitis. Our data also suggest that the amnion cell layer may provide an effective barrier against infection of the amniotic fluid. PMID- 9746600 TI - Interleukin-12 has a role in mediating resistance of murine strains to Tyzzer's disease. AB - Clostridium piliforme induces enterohepatic disease in many domestic and laboratory animal species. Susceptibility to infection is known to vary with the immune status and strain of the host, but little is known about specific immune mechanisms that regulate this disease. To evaluate host control of C. piliforme infection, we examined the role of interleukin-12 (IL-12) both in the control of and in the response to murine C. piliforme infection. For this study, 3-week-old C. piliforme-resistant C57BL/6 or -susceptible DBA/2 mice were infected intravenously with either the toxic H1 or the nontoxic M1 C. piliforme isolate. Serum and liver samples were collected prior to C. piliforme inoculation (day 0) and at days 1, 3, 7, 14, and 28 postinoculation. Evaluation of hepatic IL-12 p40 mRNA expression by reverse transcription-PCR and of total-IL-12 protein levels in serum by enzyme-linked immunosorbent assay revealed that C. piliforme induced elevations in both hepatic p40 mRNA and serum total-IL-12 levels at all times postinoculation. Elevations were similar with both toxic and nontoxic C. piliforme isolates. Levels of total IL-12 in serum were significantly (P < 0.05) higher in C57BL/6 mice than in DBA/2 mice. Additional experiments were performed in which polyclonal antibody treatment was used to neutralize IL-12 in mice of both strains prior to intravenous inoculation with toxic C. piliforme H1. IL-12 neutralization increased the severity of Tyzzer's disease at day 3 postinoculation in both mouse strains, but the degree of increase was greater in C57BL/6 mice than in DBA/2 mice. PMID- 9746601 TI - Production of interleukin-12 by murine macrophages in response to bacterial peptidoglycan. AB - Peptidoglycan (PG), a component of the bacterial cell wall, has various immunomodulating activities, including the capacity to induce delayed-type hypersensitivity reactions to antigens administered in Freund's adjuvant. We report that PG induces interleukin-12 (IL-12) mRNA production and IL-12 secretion by mouse macrophages. The capacity of PG to induce IL-12 production, like its previously reported immunomodulating activities, was dependent on the structure of its peptide subunit. PG from Bacillus megaterium and Staphylococcus aureus induced IL-12 production, whereas PG from Micrococcus luteus and Corynebacterium poinsettiae did not. The ability of most bacterial PGs to induce IL-12 production suggests that they play an important role in triggering host defense mechanisms against bacterial infections. PMID- 9746602 TI - Biological and biochemical characteristics of cytoadhesion of Plasmodium falciparum-infected erythrocytes to chondroitin-4-sulfate. AB - The cytoadhesion of Plasmodium falciparum laboratory strains and clones to Saimiri brain microvascular endothelial cells (SBEC 17), with chondroitin-4 sulfate (CSA) as the only adhesion receptor, was tested. Only one strain had significant cytoadhesion. However, CSA-specific infected erythrocytes (IRBCs) were detected in all strains after selection of a CSA-specific subpopulation by culturing the few adherent IRBCs. This demonstrates the lack of sensitivity of cytoadhesion microassays for detecting small quantities of CSA-specific IRBCs in cultures or field isolates. Cytoadhesion to CSA is maximal at 24 h of the cycle and decreases with the onset of schizogony, reaching a minimum just before reinvasion. This fluctuation must be taken into account in comparisons of the cytoadhesion of different strains or isolates. The minimum size of CSA for active inhibition was 4 kDa, and a mass of 9 kDa was required for inhibition similar to that obtained with the 50-kDa CSA. In contrast to cytoadhesion to CSA, which is pH independent or maximal at physiological pH (depending on the target endothelial cells), adhesion to CD36 and intercellular adhesion molecule 1 was pH dependent, requiring acidic conditions to be maximal in all cases. Cytoadhesion to CSA may trigger the occlusion of microvessels and cause the acidosis necessary for the other receptors to be fully efficient. If this key role in the mechanisms of sequestration were to be confirmed in vivo, prevalence studies of the CSA cytoadhesion phenotype would have to be reevaluated, because simple cytoadhesion assays do not detect CSA-specific parasites present in very low numbers, and these parasites might then be undetected in the peripheral blood but present in organs in which sequestration occurs, such as the placenta (M. Fried and P. E. Duffy, Science 272:1502-1504, 1996). PMID- 9746603 TI - Expression of the virulence plasmid-carried apyrase gene (apy) of enteroinvasive Escherichia coli and Shigella flexneri is under the control of H-NS and the VirF and VirB regulatory cascade. AB - The transcription of the virulence plasmid (pINV)-carried invasion genes of Shigella flexneri and enteroinvasive Escherichia coli (EIEC) is induced at 37 degreesC and repressed at 30 degreesC. In this work, we report that the O135: K :H- EIEC strain HN280 and S. flexneri SFZM53, M90T, and 454, of serotypes 4, 5, and 2a, respectively, produce apyrase (ATP-diphosphohydrolase), the product of the apy gene. In addition, the S. flexneri strains, but not the EIEC strain, produce a nonspecific phosphatase encoded by the phoN-Sf gene. Both apy and phoN Sf are pINV-carried loci whose contribution to the pathogenicity of enteroinvasive microorganisms has been hypothesized but not yet established. We found that, like that of virulence genes, the expression of both the apy and the phoN-Sf genes was temperature regulated. Strain HN280/32 (a pINV-integrated avirulent derivative of HN280 which has a severe reduction of virB transcription) expressed the apy gene in a temperature-regulated fashion but to a much lower extent than wild-type HN280, while the introduction of the Deltahns deletion in HN280 and in HN280/32 induced the wild-type temperature-independent expression of apyrase. These results indicated that a reduction of virB transcription, which is known to occur in the pINV-integrated strain HN280/32, accounts for reduced apyrase expression and that the histone-like protein H-NS is involved in this regulatory network. Independent spontaneously generated mutants of HN280 and of SFZM53 which had lost the capacity to bind Congo red dye (Crb-) were isolated, and the molecular alterations of pINV were evaluated by PCR analysis. Alterations of pINV characterized by the absence of virF or virB and by the presence of the intact apy locus or intact apy and phoN-Sf loci were detected among Crb- mutants of HN280 and SFZM53, respectively. While all Crb- apy+ mutants of HN280 failed to produce apyrase, Crb- apy+ phoN-Sf+ mutants of SFZM53 lacked apyrase activity but produced a nonspecific phosphatase, like parental SFZM53. Moreover, the introduction of recombinant plasmids carrying cloned virF (pMYSH6504) or virB (pBN1) into Crb- mutants of HN280 and SFZM53 lacking virF or virB, respectively, fully restored temperature-dependent apyrase expression to levels resembling those of the parental strains. Taken together, our results demonstrate that, as has already been shown for invasion genes, apy is another locus whose expression is controlled by temperature, H-NS, and the VirF and VirB regulatory cascade. In contrast, the temperature-regulated expression of the nonspecific phosphatase does not appear to be under the control of the same regulatory network. These findings led us to speculate that apyrase may play a role in the pathogenicity of enteroinvasive bacteria. PMID- 9746604 TI - Identification of two laminin-binding fimbriae, the type 1 fimbria of Salmonella enterica serovar typhimurium and the G fimbria of Escherichia coli, as plasminogen receptors. AB - Escherichia coli strains carrying recombinant plasmids encoding either the type 1 fimbria of Salmonella enterica serovar Typhimurium or the G fimbria of E. coli exhibited binding of human 125I-Glu-plasminogen and enhanced the tissue-type plasminogen activator-catalyzed conversion of plasminogen to plasmin. Purified type 1 or G fimbriae similarly bound plasminogen and enhanced its activation. The binding of plasminogen did not involve the characteristic carbohydrate-binding property of the fimbriae but was inhibited at low concentrations by the lysine analog epsilon-aminocaproic acid. Because these fimbrial types bind to laminin of basement membranes (M. Kukkonen et al., Mol. Microbiol. 7:229-237, 1993; S. Saarela et al., Infect. Immun. 64:2857-2860, 1996), the results demonstrate a structural unity in the creation and targeting of bacterium-bound proteolytic plasmin activity to basement membranes. PMID- 9746605 TI - Localization of a T-cell epitope of superantigen toxic shock syndrome toxin 1 to residues 125 to 158. AB - Toxic shock syndrome toxin 1 (TSST-1) is a member of the staphylococcal enterotoxin superantigen family. So far, little is known about T-cell epitopes on superantigens. In this study, we developed an improved method for localizing T cell epitopes on superantigens that involved synthetic peptides plus costimulation by CD28 or phorbol myristate acetate. Using this method, we localized a T-cell epitope to a 34-residue region, TSST-1 (residues 125 to 158), which possessed only two of four TSST-1-targeted beta-chain variable element (Vbeta) specificities of T-cell receptors in humans and mice, human Vbeta2 and murine Vbeta15. PMID- 9746606 TI - Plasminogen binding and activation at the surface of Helicobacter pylori CCUG 17874. AB - The binding of iodine-labelled plasminogen to Helicobacter pylori CCUG 17874 was characterized. Inhibition of the binding was observed after preincubation of H. pylori cells with nonradiolabelled plasminogen, lysine, or the lysine analogue epsilon-aminocaproic acid. Fragments of plasminogen, kringles 1 to 3, kringle 4, and mini-plasminogen, were also studied as potential inhibitors. Mini-plasminogen caused total inhibition of the plasminogen binding, while the other fragments caused only partial inhibition. These findings suggest that H. pylori binds specifically the fifth kringle structure of the plasminogen molecule. Plasminogen binding to H. pylori seems to be independent of culture media and independent of the presence of the cytotoxin-associated CagA antigen. Immunoblot analysis identified two plasminogen binding proteins of 57 and 42 kDa. Scatchard plot analysis revealed one binding mechanism with a Kd value of 7 x 10(-7) M. Conversion of H. pylori cell-bound plasminogen to plasmin in the presence of a tissue-type plasminogen activator was demonstrated by digestion of the chromogenic substrate S-2251. No activation was noted when plasminogen or tissue type plasminogen activator was incubated with H. pylori cells alone. Formation of H. pylori cell surface-bound plasmin may be important to provide a powerful proteolytic mechanism for gastric tissue penetration in type B gastritis and peptic ulcer disease, since plasmin degrades not only fibrin but also extracellular matrix proteins such as various collagens and fibronectin. PMID- 9746607 TI - An ex vivo study of T lymphocytes recovered from the lungs of I/St mice infected with and susceptible to Mycobacterium tuberculosis. AB - I/St mice, previously characterized as susceptible to Mycobacterium tuberculosis H37Rv, were given 10(3) or 10(5) CFU intravenously. At two time points postinoculation, the cell suspensions that resulted from enzymatic digestion of lungs were enumerated and further characterized phenotypically and functionally. Regarding the T-cell populations recovered at 2 and 5 weeks postinfection, two main results were obtained: (i) the population of CD44(-) CD45RB+ cells disappeared within 2 weeks postinfection, while the number of CD44(+) CD45RB-/low cells slowly increased between weeks 2 and 5; (ii) when cocultured with irradiated syngeneic splenocytes, these lung T cells proliferated in the presence of H37Rv sonicate. Using H37Rv sonicate and irradiated syngeneic splenocytes to reactivate lung T cells, we selected five CD3(+) CD4(+) CD8(-) T-cell clones. In addition to the H37Rv sonicate, the five clones react to both a short-term culture filtrate and an affinity-purified 15- to 18-kDa mycobacterial molecule as assessed by the proliferative assay. However, there was a clear difference between T-cell clones with respect to cytokine (gamma interferon [IFN-gamma] and interleukin-4 [IL-4] and IL-10) profiles: besides one Th1-like (IFN-gamma+ IL-4( )) clone and one Th0-like (IFN-gamma+ IL-4(+) IL-10(+)) clone, three clones produced predominantly IL-10, with only marginal or no IL-4 and IFN-gamma responses. Inhibition of mycobacterial growth by macrophages in the presence of T cells was studied in a coculture in vitro system. It was found that the capacity to enhance antimycobacterial activity of macrophages fully correlated with INF gamma production by individual T-cell clones following genetically restricted recognition of infected macrophages. The possible functional significance of cytokine diversity among T-cell clones is discussed. PMID- 9746608 TI - Effect of cytokines on growth of Toxoplasma gondii in murine astrocytes. AB - Cytokines play a significant role in the regulation of Toxoplasma gondii in the central nervous system. Cytokine-activated microglia are important host defense cells in central nervous system infections. Recent evidence indicates that astrocytes can also be activated by cytokines to inhibit intracellular pathogens. In this study, we examined the effect of gamma interferon (IFN-gamma), tumor necrosis factor alpha (TNF-alpha), interleukin-6 (IL-6), and IL-1 on the growth of T. gondii in a primary murine astrocyte culture. Pretreatment of astrocytes with IFN-gamma resulted in 65% inhibition of T. gondii growth. Neither TNF-alpha, IL-1, nor IL-6 alone had any effect on T. gondii growth. IFN-gamma in combination with either TNF-alpha, IL-1, or IL-6 caused a 75 to 80% inhibition of growth. While nitric oxide was produced by astrocytes treated with these cytokines, inhibition of T. gondii growth was not reversed by the addition of the nitric oxide synthase inhibitor NG-monomethyl-L-arginine. Furthermore, IFN-gamma in combination with IL-1, IL-6, or TNF-alpha also induced inhibition in astrocytes derived from syngeneic mice deficient in the enzyme inducible nitric oxide synthase. This finding suggests that the mechanism of cytokine inhibition is not nitric oxide mediated. Similarly, the addition of tryptophan had no effect on inhibition, indicating that the mechanism was not mediated via induction of the enzyme indoleamine 2, 3-dioxygenase. The mechanism of inhibition remains to be elucidated. Results from this study demonstrate that cytokine-activated astrocytes are capable of significantly inhibiting the growth of T. gondii. These data indicate that astrocytes may be important host defense cells in controlling toxoplasmosis in the brain. PMID- 9746609 TI - Interleukin-12 is essential for a protective Th1 response in mice infected with Cryptococcus neoformans. AB - To analyze the roles of interleukin-12 (IL-12) and the IL-12-dependent Th1 response in resistance to Cryptococcus neoformans, we have established a chronic infection model in wild-type mice and in mice with targeted disruptions of the genes for the IL-12p35 and IL-12p40 subunits (IL-12p35(-/-) and IL-12p40(-/-) mice, respectively) as well as in mice with a targeted disruption of the IL-4 gene. Long-term application of exogenous IL-12 prevented death of infected wild type mice for the entire period of the experiment (up to 180 days) but did not resolve the infection. Infected IL-12p35(-/-) and IL-12p40(-/-) mice died significantly earlier than infected wild-type mice, whereas infection of IL-4 deficient mice led to prolonged survival. Interestingly, infected IL-12p40(-/-) mice died earlier and developed higher organ burdens than IL-12p35(-/-) mice, which, for the first time in an infection model, suggests a protective role of the IL-12p40 subunit independent of the IL-12 heterodimer. The fungal organ burdens of IL-4-deficient mice and IL-12-treated wild-type mice were significantly reduced compared to those of untreated wild-type mice and IL-12 deficient mice. Histopathological analysis revealed reduction of the number of granulomatous lesions following treatment with IL-12. Susceptibility of both IL 12p35(-/-) and IL-12p40(-/-) mice was associated with marginal production of gamma interferon and elevated levels of IL-4 from CD4(+) T cells, which indicates Th2 polarization in the absence of IL-12, whereas wild-type mice developed a Th1 response. Taken together, our data emphasize the essential role of IL-12 for protective Th1 responses against C. neoformans. PMID- 9746610 TI - Infection of epithelial cells by pathogenic neisseriae reduces the levels of multiple lysosomal constituents. AB - Members of our group reported recently that neisseria infection of human epithelial cells results in accelerated degradation of the major lysosomal integral membrane protein LAMP1 and that this is due to hydrolysis of this glycoprotein at its immunoglobulin A1 (IgA1)-like hinge by the neisseria type 2 IgA1 protease (L. Lin et al., Mol. Microbiol. 24:1083-1094, 1997). We also reported that the IgA1 protease plays a major role in the ability of the pathogenic neisseriae to survive within epithelial cells and hypothesized that this is due to alteration of lysosomes as a result of protease-mediated LAMP1 degradation. In this study, we tested the hypothesis that neisseria infection leads to multiple changes in lysosomes. Here, we report that neisseria infection also reduces the levels of three other lysosomal markers: LAMP2, lysosomal acid phosphatase (LAP), and CD63. In contrast, neither the epidermal growth factor receptor level nor the beta-tubulin level is affected. A detailed examination of LAMP2 indicated that the reduced LAMP2 levels are not the result of an altered biosynthetic rate or of cleavage by the IgA1 protease. Nevertheless, the protease plays a role in reducing LAMP2 and LAP activity levels, as these are partially restored in cells infected with an iga mutant. We conclude that neisseria infection results in multiple changes to the lysosomes of infected epithelial cells and that these changes are likely an indirect result of IgA1 protease mediated cleavage of LAMP1. PMID- 9746611 TI - The cell cycle-specific growth-inhibitory factor produced by Actinobacillus actinomycetemcomitans is a cytolethal distending toxin. AB - Actinobacillus actinomycetemcomitans has been shown to produce a soluble cytotoxic factor(s) distinct from leukotoxin. We have identified in A. actinomycetemcomitans Y4 a cluster of genes encoding a cytolethal distending toxin (CDT). This new member of the CDT family is similar to the CDT produced by Haemophilus ducreyi. The CDT from A. actinomycetemcomitans was produced in Escherichia coli and was able to induce cell distension, growth arrest in G2/M phase, nucleus swelling, and chromatin fragmentation in HeLa cells. The three proteins, CDTA, -B and -C, encoded by the cdt locus were all required for toxin activity. Antiserum raised against recombinant CDTC completely inhibited the cytotoxic activity of culture supernatant and cell homogenate fractions of A. actinomycetemcomitans Y4. These results strongly suggest that the CDT is responsible for the cytotoxic activity present in the culture supernatant and cell homogenate fractions of A. actinomycetemcomitans Y4. This CDT is a new putative virulence factor of A. actinomycetemcomitans and may play a role in the pathogenesis of periodontal diseases. PMID- 9746612 TI - Mutational analysis of superantigen activity responsible for the induction of skin erythema by streptococcal pyrogenic exotoxin C. AB - Streptococcal pyrogenic exotoxin C (SPEC), when injected intradermally, induces erythema in unsensitized rabbits. In the present study, we examined whether this erythema induction is due to the T-cell stimulatory activity of SPEC as a superantigen. Analysis by using single-residue mutant SPECs indicated that mutant SPECs Y15I, A16E, and Y17I, in which tyrosine 15, alanine 16, and tyrosine 17 were replaced with isoleucine, glutamic acid, and isoleucine, respectively, exhibited significantly reduced mitogenic activity for Vbeta2(+) human T cells in vitro, and Y15I showed as much as a 1, 000-fold reduction. Y15I mutant SPEC, however, retained the ability to bind to major histocompatibility complex class II antigen and to form a homodimer, implying that residue 15 is critically important for the interaction of SPEC with T-cell antigen receptor beta chains. When injected intradermally into normal rabbits, wild-type SPEC induced a characteristic erythema after 3 h in a dose-dependent fashion, which was associated with polymorphonuclear and mononuclear cell infiltration. This erythema formation was found to be severely suppressed by systemic pretreatment with cyclosporin A, suggesting the involvement of host T cells. Y15I mutant SPEC exhibited nearly 1, 000-fold less erythema induction in vivo than wild-type SPEC. Altogether, the present results strongly suggest that erythema induction in rabbits by SPEC is attributable mostly to its T-cell stimulatory activity as a superantigen. PMID- 9746613 TI - Organ-dependent variation of capsule thickness in Cryptococcus neoformans during experimental murine infection. AB - In studies of murine infection, the capsule thickness of Cryptococcus neoformans varied depending on the organ. The relative order of thickness was as follows: lung > brain > in vitro isolates. The differences in capsule thickness suggest that there are organ-related differences in the expression of genes responsible for capsule thickness. PMID- 9746614 TI - Pathogenicity of an enterotoxigenic Escherichia coli hemolysin (hlyA) mutant in gnotobiotic piglets. AB - Pigs infected with hemolytic F4(+) strains of enterotoxigenic Escherichia coli often develop septicemia secondary to intestinal infection. We tested the hypothesis that inactivation of hemolysin would reduce the ability of F4(+) enterotoxigenic E. coli to cause septicemia in swine following oral inoculation. Inactivation of the hemolysin structural gene (hlyA) did not decrease the incidence of septicemia in the gnotobiotic piglet model. PMID- 9746615 TI - CD95 expression in aged mice infected with tuberculosis. AB - The interaction between CD95 and its ligand is an important homeostatic mechanism that leads to the induction of apoptosis in activated T cells. In view of recent evidence that this pathway might be defective in aged mice, this study investigated CD95 expression on T cells in old mice activated by infection with Mycobacterium tuberculosis. The results of the study do not support the hypothesis that CD95 is poorly expressed on CD4 T cells from old mice; instead, it was found that similar numbers of T cells from young and old mice expressed CD95, with the intensity of expression if anything higher on the cells from the old mice. In addition, the study demonstrated that changes in CD44 and CD45RB expression previously observed in young infected mice proceeded in a similar fashion in old animals and, as would be predicted, that CD95(hi) expression was primarily associated with CD4 T cells expressing the activated CD44(hi) CD45RBhi phenotype. PMID- 9746618 TI - Management of Abnormal Cervical/Vaginal Pap Smears. AB - The primary role of the Pap smear is to detect premalignant lesions. Unfortunately, it is unclear how best to manage patients with low-grade lesions or squamous atypia. There are simple approaches to common problems seen with Pap smears, such as infection, inflammation, and no endocervical cells on the smear. PMID- 9746617 TI - Effective, nonsensitizing vaccination with culture filtrate proteins against virulent Mycobacterium bovis infections in mice. AB - Vaccination of mice with Mycobacterium bovis culture filtrate proteins (CFP), prepared in a variety of adjuvants (aluminum hydroxide, Quil-A, and dimethyldioctyldecyl ammonium bromide [DDA]), provided significant protection against an aerosol challenge of virulent M. bovis. Additionally, vaccination with CFP in DDA or Quil-A did not sensitize mice to M. bovis purified protein derivative. PMID- 9746619 TI - A Revised Strategy for the Prevention of Group B Streptococcal Infection in Pregnant Women and Their Newborns. AB - Group B beta-hemolytic Streptococcus (GBS) is the leading cause of life threatening perinatal infection of newborns in developed countries. Because a vaccine is not yet available, selective intrapartum chemoprophylaxis is the best current strategy for preventing disease. Joint recommendations of the Centers for Disease Control and Prevention (CDC), the American College of Obstetricians and Gynecologists (ACOG), and the American Academy of Pediatrics (AAP) are that all pregnant women be screened for GBS at 35 to 37 weeks of gestation. Pregnant women who are colonized with GBS should be treated with intravenous penicillin during labor. Women who have not been screened but exhibit risk factors known to be associated with GBS disease, such as preterm labor and/or membrane rupture at fewer than 37 weeks' gestation, intrapartum fever, and prolonged rupture of membranes > 18 hours, should also receive intrapartum antibiotics if they begin labor. Women with a history of GBS disease, such as a prior episode of GBS bacteriuria or a previous newborn with invasive GBS disease, are at high risk for recurrent GBS infection. The latter 2 categories in particular warrant chemoprophylaxis regardless of colonization status. PMID- 9746616 TI - Acquisition of iron by Gardnerella vaginalis. AB - Six Gardnerella vaginalis strains were examined for the ability to utilize various iron-containing compounds as iron sources. In a plate bioassay, all six strains acquired iron from ferrous chloride, ferric chloride, ferrous sulfate, ferric ammonium citrate, ferrous ammonium sulfate, bovine and equine hemin, bovine catalase, and equine, bovine, rabbit, and human hemoglobin. All six strains also acquired iron from human lactoferrin, but not from human transferrin, as determined by a liquid broth growth assay. Siderophore production was detected in eight G. vaginalis strains by the chrome azurol S universal chemical assay. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the cytoplasmic membrane proteins isolated from G. vaginalis 594 grown under iron replete and iron-restricted conditions revealed several iron-regulated proteins ranging in molecular mass from 33 to 94 kDa. These results indicate that G. vaginalis may acquire iron from iron salts and host iron compounds. PMID- 9746620 TI - Evaluation of Female Urinary Incontinence. AB - History and physical examination, along with simple diagnostic tests, are the best means for evaluating urinary incontinence. History assesses the severity of the incontinence and its effect on the patient's life. Physical examination is necessary to evaluate the degree of incontinence and possible neurologic disorders. A cystometrogram can be used to evaluate bladder compliance. It is easy to perform and may be the most important and useful urodynamic test for this problem. PMID- 9746621 TI - Management of Asymptomatic UTIs in Women. AB - Asymptomatic bacteriuria is common in women and increases markedly with age and functional impairment. Although asymptomatic bacteriuria frequently resolves spontaneously, it may persist for extended periods, particularly in older women. Programs to screen for and treat asymptomatic bacteriuria are appropriate only in pregnant women. For the general female population, screening and treatment programs for asymptomatic bacteriuria do not appear warranted. PMID- 9746622 TI - Kikuchi-Fujimoto Disease: An Exuberant Localized T Cell Activation Arrested By Histiocytes? AB - Kikuchi-Fujimoto disease, or histiocytic necrotizing lymphadenitis, is a relatively new disease entity seen in the U.S. that primarily affects young women. Originally discovered in 1972 in Japan, it is now seen in the U.S. By 1993 approximately 200 cases had been reported. The disease is a self-limiting process of unknown etiology. PMID- 9746623 TI - Emergency Contraception: More Than A Morning After Pill. AB - Emergency contraception (postcoital contraception, the "morning-after pill") has been available for almost 30 years but remains vastly underutilized. As many as 50% of undesired pregnancies could be prevented with the use of emergency contraception. Currently used regimens include ethinyl estradiol/norgestrel (the Yuzpe regimen) and copper-containing IUDs. The limiting side effects with the Yuzpe regimen are nausea and vomiting. Potential agents of the future include mifepristone and levonorgestrel, which do not include estrogen and thereby minimize nausea and vomiting. More widespread education of physicians and patients about the safety and benefits of emergency contraceptive treatment is necessary. PMID- 9746624 TI - Abdominal Tuberculosis or Ovarian Carcinoma: Management Dilemma Associated With an Elevated CA-125 Level. AB - The CA-125 antigen is useful for detecting residual disease in women treated for ovarian cancer. Its role in screening for ovarian cancer is questionable, however, because anumber of other benign and malignant causes can elevate its level. In this case, a patient with genital tract tuberculosis was initially misdiagnosed with ovarian cancer. PMID- 9746625 TI - Infection After Gynecologic Surgery: Implications of Racial Leukopenia in Black Women. AB - Differences in the normal peripheral white blood cell count (WBC) among people of various racial backgrounds are well recognized, and one familiar example is the decrease in the WBC of African-Americans relative to that of Caucasian subjects, a phenomenon that has been termed "racial leukopenia." Since the WBC is commonly used as an early indicator of infection, information regarding this phenomenon is pertinent: Such leukopenia could give rise to a falsely low (i.e., in the normal range) WBC during a significant infectious event. We therefore conducted a retrospective study to describe the incidence of this leukopenia among Black women in Louisiana and to follow their WBC levels in response to infection after elective gynecologic surgery. We found that 6.5% of these women exhibited a preoperative WBC < 3.5 x 10^3 cells/mm3, which represents the lower limit of normal for this measure. Although these women did exhibit a 2- to 3-fold rise in their WBC in response to infection, their peak WBC remained less than 11.0 x 10^3/mm3, a value below the upper limit of normal. PMID- 9746626 TI - Contemporary Strategies for Detecting Chlamydial Infection in Women. AB - Chlamydia trachomatis infections are the most common bacterial cause of sexually transmitted disease in the US. In women, these infections often result in such serious reproductive tract complications as pelvic inflammatory disease, infertility, and ectopic pregnancy, and an infected woman can pass the infection to her newborn during delivery. The pervasiveness of this often asymptomatic disease necessitates that health care providers actively look for C trachomatis infection, especially in young women. The diagnosis of chlamydial infections has historically been difficult, but newer chlamydia diagnostic tests have become clinically available in the past decade. PMID- 9746627 TI - Seeking A Rational Approach to the Diagnosis and Treatment of Interstitial Cystitis. AB - At least 90% of the half-million Americans with interstitial cystitis (IC) are women. But correctly diagnosing and treating the problem are not as clear as the statistics. The clinical hallmarks--bladder pain, urinary urgency and frequency, nocturia, and dysuria--are not confined to IC. A key feature of diagnosing the problem is to exclude the vast array of other possibilities, including carcinoma and detrusor hyperreflexia. To confirm the diagnosis, cystoscopic exam with hydrodistention is needed to visualize diffuse glomerulations or classic Hunner's ulcer. Treatment is empiric and can involve a variety of oral and intravesical therapies. Patients with a bladder capacity less than 250cc may benefit from surgery. PMID- 9746628 TI - Update on Hormone Replacement: Sorting Out the Options for Preventing Coronary Artery Disease and Osteoporosis. AB - Menopause is associated with an increased risk of osteoporosis and coronary artery disease. The onset of menopause is an excellent time to assess a woman's overall health and the need for hormone replacement therapy (HRT) or other measures to reduce menopausal symptoms, prevent bone loss, and reduce the risk of heart attack. Available HRT can be given in a variety of dosages and formulations, and it can be adjusted to the needs of the individual. If HRT is contraindicated or not accepted, 2 alternative therapies (alendronate and salmon calcitonin) may be offered to treat osteoporosis. PMID- 9746629 TI - Critical Issues in Treating HIV During Pregnancy. AB - When the AIDS Clinical Trial Group (ACTG) broke the code for Protocol 076, researchers discovered that giving zidovudine (formerly called azidothymidine, or AZT) to HIV-infected women during pregnancy (100mg PO 5 times daily) and labor (2mg/kg of body weight IV, given over 1 hour, then 1mg/kg/hr until delivery), plus to the newborn (2mg/kg PO q6h for 6 weeks), could cut perinatal HIV transmission by two thirds. Now, further studies are also adding to the databank of information about HIV in pregnancy. For example, one study has shown that pregnancy does not hasten the progression of HIV; another has indicated that although vertical HIV transmission often occurs late in gestation, it also can occur as early as 8 weeks. One group of researchers discovered that zidovudine is highly effective in blocking viral transmission, but when the mother develops resistance, she is likely to pass the resistant viral strain to the fetus. The importance of measuring viral load has been suggested by research demonstrating that the number of HIV RNA copies can help predict which women are most likely to transmit the virus to the fetus. In one study, among women with more than 80,000 RNA copies/mL, everybody transmitted the virus; with less than 20,000 copies, nobody transmitted. Of great interest is that the 22 women in this study who took zidovudine showed an 8-fold median decrease in the plasma RNA levels. PMID- 9746630 TI - Recognizing and Intervening in Domestic Violence: Proactive Role for Dentistry. AB - Violence within families and in intimate relationships is now recognized as a widespread problem. Abuse tends to recur and may continue until the victim dies as a result of his or her injuries. Every effort should be made to intervene and stop this chain of events so that the cycle of violence will cease. Although victims are often reluctant to report their abuse, the fac that about 75% of physical injuries are inflicted to the region of the head, face, mouth, and neck places dentists in a particularly good position to recognize the signs of abuse and take steps to intervene. In this article, the signs and symptoms of physical, emotional, and sexual abuse are discussed, and the proactive steps that dental health professionals can take to record and report these cases are outlined. PMID- 9746631 TI - Minimizing the Risk of Delayed Diagnosis of Breast Cancer. AB - According to the Physician Insurers Association of America (PIAA), the most expensive and common medicolegal claim against physicians related to errors of diagnosis in malignant diseases is delay in the diagnosis of breast cancer. Although just 40% of all breast cancers occur in women younger than 50 years old, 60% of the claims of a delayed diagnosis of breast cancer arise in this group. Unimpressive findings on a physical exam (54.7%) topped the list of reasons for physician-related causes of delay in diagnosing breast cancer. Other factors in the PIAA survey included negative mammogram report (35.7%), failure to do an appropriate biopsy to evaluate a suspicious mass (26.8%), no suspicious findings discovered on repeat exam (24.5%), delay in requesting a consult or in referring a patient for further evaluation (18.2%), record-keeping errors (16.0%), inadequate physician-to-physician communication (13.8%), physician distraction by patient's other health problems (12.3%), mammogram misread (8.8%), and poor physical examination done by physician (5.9%). Three elements are vital to an effective breast-screening program: a comprehensive breast history, a thorough breast examination, and a clear record of findings and follow-up. PMID- 9746632 TI - Autologous Stem Cell Transplants Taking Breast and Ovarian Cancer Chemotherapy to New Heights. AB - Traditional chemotherapy regimens for advanced breast cancer have produced complete response (CR) rates of 5%-25% and remissions of 5-13 months. Autologous stem cell transplantation (ASCT), however, permits a 5- to 10-fold escalation of chemotherapy, which results in a substantial increase in tumor-cell killing relative to standard therapy. This technique involves protecting hematopoietic function by collecting stem cells from bone marrow or peripheral blood before subjecting the patient to high-dose chemotherapy. These stem cells are then reinfused in an autologous bone marrow transplant (ABMT) or autologous blood stem cell transplant (ABSCT) to quickly repopulate the patient's marrow and blood. The largest and most mature of phase II trials of ASCT for metastatic breast cancer reports rates of 35%-68% and 4-year survival rates of 19%-27%. Although most patients develop recurrent disease 6-12 months post-ASCT, it is encouraging that a few patients survive, disease-free, beyond 5 years. The Autologous Blood and Marrow Transplant Registry analyzed data on more than 800 patients with metastatic breast cancer who underwent ASCT. The Registry reported a 3-year progression-free survival rate of 20% and an overall survival rate of 37%, which compares favorably with an expected 17%-25% 3-year survival rate after conventional chemotherapy. Whether similar success can be achieved with ASCT in ovarian cancer is uncertain because few ovarian cancer patients are presently entered in high-dose studies using ASCT support. PMID- 9746633 TI - Maternal Serum Markers of Fetal Abnormalities: Progress in Prenatal Screening. AB - Second-trimester screening of a pregnant woman's serum for markers of such congenital abnormalities as fetal trisomy 21 (Down syndrome) and neural tube defects has become an important part of antenatal care. The protocol most commonly used is the triple marker screen, which measures serum levels of alpha fetoprotein (AFP), unconjugated estriol (uE3), and beta-human chorionic gonadotropin (beta-hCG). The results of these tests are used to derive a value known as a multiple of the median (MoM). An algorithm that combines maternal age with MoMs can be used to calculate the risk for such chromosomal disorders as trisomy 21 and 18. AFP alone--corrected for weight, race, insulin-dependent diabetes, and multiple pregnancy--can be used to predict the risk of fetal neural tube defect. Prenatal screening is best performed within an integrated program providing biochemical testing in conjunction with knowledgeable experienced counselors, expert ultrasonographers, and physicians with special expertise in maternal fetal medicine and neonatal care. Prenatal screening provides parents with the information they need to make informed pregnancy decisions and, if necessary, organize support and resources essential to the care of an infant with special needs. PMID- 9746634 TI - Assessing the Mental Health Impact of Induced Abortion. AB - Induced abortion is the subject of tremendous political conflict. Anti-abortion activists have frequently asserted that abortion results in major psychiatric sequelae. The evidence, however, clearly indicates that this is not necessarily the case. Studies performed to date confirm that it is possible to identify risk factors, help pregnant women make decisions compatible with their own values, and minimize psychiatric sequelae. PMID- 9746635 TI - Why the Urgency for Earlier Diagnosis of HIV Infection in Women? AB - As our perspective of AIDS changes from a disease that affects homosexual men and intravenous drug abusers to one that spreads increasingly through heterosexual contact, greater numbers of women are being diagnosed with HIV infection. Because clinicians still may perceive AIDS to be primarily a "male" disease and may maintain a low level of suspicion of it in their female patients, HIV infection is believed to be underdiagnosed and go unrecognized for longer periods in women than in men. Early reports in the 1980s that showed a worse prognosis and shorter median survival for women than for men are now explained by factors such as later presentation and diagnosis, disparity in access to care, and reluctance of clinicians to prescribe zidovudine to pregnant patients. Recent studies show that, with early identification of infection and subsequent treatment and prophylactic therapy, there are no differences in disease progression or survival in women compared with men. Moreover, from a public health standpoint, earlier diagnosis can decrease incidence of both sexual and vertical transmission. Women may choose to undergo tubal ligation to prevent future vertical transmission; additionally, zidovudine now is used in pregnant women and has been demonstrated to decrease transmission rates from 25% to 8% PMID- 9746636 TI - Critical Challenges of Renal Calculi in Women. AB - Stone characteristics, patient factors, and technological features must all be weighted before selecting optimal therapy. A key patient factor is gender. Although renal calculi generally are more frequent in men than in women, the number of women who suffer from urolithiasis has been on the rise in the past 10 years. Calcium and uric acid stones are formed mainly in men; potentially dangerous struvite stones are more common in women. Renal stones in women who are pregnant or even just in their reproductive years can present complex treatment challenges. When examining stone factors, key considerations are the location and size of the stone. For renal calculi smaller than 2cm and located in the upper urinary tract, extracorporeal shock wave lithotripsy (ESWL) is often an effective way to clear the stone, as long as the patient is not pregnant. For larger stones, percutaneous nephrostolithotomy (PNL) may be used to access and fragment the stone using contact (ultrasonic, electrohydraulic, or laser) lithotripsy. For ureteral stones smaller than 5mm that fail to pass spontaneously, ESWL or uteteroscopy can be an effective means to remove them. ESWL and ureteroscopy are currently the principal interventions used in ureteral stone management. PMID- 9746637 TI - Recognizing the Range of Mood Disorders in Women. AB - Depression strikes twice as many women as men. The dramatic changes in psychoactive hormones like estrogen and progesterone that occur during menstrual cycles and during pregnancy are believed to be key factors. According to the 4th Edition of the Diagnostic and Statistical Manual (DSM-IV), the diagnosis of depression requires that specific criteria be met: depressed mood or loss of interest or pleasure of at least 2 weeks' duration plus at least 4 of the following additional symptoms: change in weight, altered sleep pattern, psychomotor agitation or retardation, fatigue, difficulty concentrating, feelings of worthlessness or guilt, and suicidal ideation. Several new drug therapies are available for treating major depressive illness. PMID- 9746638 TI - Can Diet and ExerciseReally Change Metabolism? AB - Resting metabolic rate (RMR; kcal/min) accounts for approximately 60% to 75% of total daily energy expenditure. The energy required to digest and absorb food, also called the thermic effect of feeding, provides an additional 5% to 10%, and the energy expended in physical activity (thermic effect of activity) comprises approximately 20% to 30% of daily energy output. The major determinants of RMR include such factors as body surface area, age, gender, thyroid hormones and catecholamines, genetics, body temperature, and in women, the phase of the menstrual cycle. Most factors are beyond the control of the individual, yet it is believed that voluntary behaviors, specifically dietary intake and exercise, can influence resting metabolism--but for how long and by how much is not well defined. In general, it is widely believed that dieting decreases metabolic rate and exercise increases it. However, present research suggests that any change may be short-term. Also, the relationship of diet, exercise, and metabolism is not a simple one. Very restrictive diets can result in a transient decrease in metabolic rate. Whether this decrease is sustained has not been clearly shown. Many believe that a permanently reduced metabolism is the reason that diets do not work and that weight regain after weight loss seems inevitable. Current research efforts are focused on ways to maintain or increase metabolic rate through diet and exercise. PMID- 9746639 TI - Managing Change: Psychosocial Issues for Women. AB - Changes in the environment can threaten human survival. Adapting to changing times has become a major issue in the late 20th century because of the drastic and frequent alterations in human life styles. Managing change in the environment depends on developing an awareness of the contextual issues that support or threaten human existence. This paper reviews the experience of a young mother who was isolated from other mothers by the urban setting in which she lived. She was able to organize a child-care cooperative, a "village" to raise the children. Through the cooperative she not only overcame her own feelings of anger and isolation, but also created a wholesome setting that supported the growth of her whole family. Such strategies of "empowered collaboration" have great potential for women around the world who need to modify the environment in order to live better lives. PMID- 9746640 TI - Contraceptive Options for Perimenopausal Women. AB - A number of safe and effective contraceptive choices exist for the perimenopausal woman. With increasing experience, we have learned that hormonal methods are safer than initially thought, particularly with the lower-dose formulations currently prescribed. Furthermore, the pill offers many noncontraceptive health benefits. And while progestins, in contraceptive doses, have a slight adverse impact on the lipoprotein profile, there has been no parallel increase in clinical cardiovascular disease. Since elimination of the Dalkon shield, the IUD has been recognized as a very safe and effective method of contraception; the risk of associated infection seems primarily related to the insertion procedure. Barrier methods provide somewhat reliable contraception, with varying degrees of added protection against STD. The decreased fertility rate among perimenopausal women allows some latitude when contraceptive methods with suboptimal efficacies are selected; however, unintended pregnancy is a significant problem for women older than 40 years, should it occur. Surgical sterilization remains the most popular form of contraception among perimenopausal-aged couples. Natural family planning techniques are safe and somewhat underutilized. All women should be counseled about the availability of postcoital contraceptive methods. PMID- 9746641 TI - Selecting the Best Surgical Option for Stress Urinary Incontinence. AB - Although there are many surgical procedures available to treat urinary incontinence, obtaining the best results depends on a thorough preoperative evaluation of the patient. Traditional urodynamic evaluation of stress urinary incontinence (SUI) focusing on maximum urethral closing pressures has been found to be less useful than the abdominal leak point pressure (ALPP) test in detecting intrinsic sphincter deficiency (ISD). An ALPP less than or equal to 60cm H2O indicates a significant degree of ISD, whereas a leak point pressure greater than or equal to 90cm H2O is usually associated with pure urethral hypermobility. When combined with the history, physical examination, and a filling cystometrogram, the additional objective data obtained from ALPP permit an accurate classification of the stress incontinence and are useful to the clinician in choosing the most appropriate treatment. Anterior colporrhaphy is still commonly used by gynecologists to treat SUI, but the narrow indications (type I SUI only) and poor long-term results have decreased its popularity. It is a poor choice for treating SUI related to urethral hypermobility. Retropubic urethropexy is the treatment standard for SUI procedures against which all other procedures have been compared. Five-year cure rates are in the range of 80% to 90%. Other procedures for treating SUI related to urethral hypermobility include laparoscopic bladder neck suspension, abdominal paravaginal repair, and needle bladder neck suspension. Surgical treatment for ISD differs from that for urethral hypermobility and includes pubovaginal sling procedures, injectable agents, and insertion of the artificial urinary sphincter. Concurrent pelvic prolapse conditions should be treated simultaneously and may dictate the surgical approach. When the proper procedure is utilized, good long-term results can be expected. PMID- 9746642 TI - Breastfeeding: Unraveling the Mysteries of Mother's Milk. AB - Most of the major progress in understanding the unique and complex features of human breast milk has emerged in just the past 2 decades. Since the late 1970s, key research has examined such aspects as the composition of breast milk, effects of maternal and environmental factors on human milk, and the effect of human milk on the infant, including the protection against disease that breast milk can confer on the newborn. The composition of human breast milk includes growth factors, hormones, enzymes, and other substances that are immune-protective and foster proper growth and nutrition in the newborn. Research suggests that lactation is robust and that a mother's breast milk is adequate in essential nutrients, even when her own nutrition is inadequate. Mature breast milk usually has constant levels of about 7g/dL carbohydrate and about 0.9g/dL proteins. But the composition of fats essential for neonatal growth, brain development, and retinal function varies according to a woman's intake, the length of gestation, and the period of lactation. Vitamins and minerals also vary according to maternal intake. But even when these nutrients are lower in breast milk than in formulas, their higher bioactivity and bioavailability more nearly meet the complete needs of neonates than do even the best infant formulas. Also, in many instances human milk components compensate for immature function, such as a neonate's inability to produce certain digestive enzymes, immunoglobulin A (IgA), taurine, nucleotides, and long-chain polyunsaturated fatty acids. PMID- 9746643 TI - What's New in the Treatment of Anorexia Nervosa and Bulimia? AB - The 3 eating disorders--anorexia nervosa, bulimia nervosa, and binge eating- occur at a frequency far greater than usually realized. Anorexia has been found to be present in up to 1% of teenage and young adult women, whereas estimates of bulimia have ranged up to 5%. The prevalence of binge-eating disorder is not known, but may be higher than bulimia. Anorexia nervosa is characterized by weight loss, body image disturbance, and a morbid fear of weight gain. Bulimia nervosa is characterized by binge eating and compensatory purging by vomiting; use of laxatives, diuretics, or diet pills; exercise; or fasting. Binge-eating disorder is characterized by binge behavior and loss of control of food intake, with an absence of purging. Eating disorders create significant mortality and morbidity. Medical complications associated with anorexia are those related to malnutrition and semistarvation. Medical complications associated with bulimia are those related to electrolyte imbalance and the physical effects of vomiting. The mortality rate of eating disorders may be as high as 15%, including deaths from arrhythmia, gastric hemorrhaging, and suicide. The core struggle within women with anorexia is the "2 Ps": feeling powerless and striving towards perfectionism. The core conflict within a bulimic person appears to be the "2 Ds": deprivation and dependency. The treatment of eating disorders includes psychotherapy, and, frequently, psychopharmacologic intervention. The psychotherapy primarily addresses issues of chaotic eating, hunger, inadequate caloric intake, conditioned response, and profound fear of expressing impulses and feelings, especially those of anger and sadness. Antidepressants, especially serotonergic agents, have been found to be useful, particularly in the treatment of bulimia. PMID- 9746644 TI - Can a Silent Kidney Infection or Genetic Predisposition Underlie Recurrent UTIs? AB - Complications resulting from persistent and repeated urinary tract infections (UTIs) account for nearly 1 million hospital admissions annually. Cystitis, a localized bladder infection occurring in the lower tract, is recognized by a symptom complex of dysuria, frequency, urgency, and suprapubic tenderness; pyelonephritis, which refers to upper tract infection of the kidneys, classically manifests with flank pain and systemic as well as cystitis signs. An empiric 3 day antibiotic regimen has been shown to be more than 95% effective in curing cystitis. But for a subgroup of patients, a relapse of "cystitis" within 4 weeks can signal a subclinical, "silent," pyelonephritis. A 14-day course of antibiotics is indicated to treat the recurrent UTI. Follow-up urinalysis and urine cultures are then repeated 2 and 4 weeks after therapy. If symptoms and/or bacteriuria are again documented with the same organism, subclinical pyelonephritis is presumed; a prolonged 6-week course of antibiotics is then warranted to prevent prolonged problems and complications associated with UTIs. When the problem is reinfection with a microorganism different from that responsible for the last infection, short-course therapy for 3 days may be prescribed for each episode. When reinfection occurs more frequently than 2 to 3 times a year, however, antibiotic prophylaxis to prevent reinfections is warranted. PMID- 9746645 TI - Innovations in Pap Screening for Cervical Neoplasia. AB - The Papanicolaou (Pap) smear has been credited with reducing deaths from cervical cancer over recent decades. Retrospective studies have shown that 49% to 72% of patients with cervical cancer had not been screened or had been improperly screened, but 30% to 50% of women with cervical cancer had had a normal smear within the past 3-4 years. About 25% of the estimated 8000 false-negative Paps a year are potentially preventable by refinement of Pap readings and an additional 25% by improvement in the sampling technique/process. On the other hand, an equal number of cases of cancer will not be detected by the Pap smear. Two innovations have been introduced to improve Pap screening: speculoscopy and automated re readings of negative Pap smears. Technical improvements in Pap smears, however, may result in only a limited improvement in mortality from cervical cancer. Improving patient compliance with screening, plus education to reduce behaviors that raise the risk of cancer, may have more of an impact on the mortality rate. PMID- 9746646 TI - Skin-Sparing Mastectomy and Immediate Breast Reconstruction. AB - Skin-sparing mastectomy refers to a standard mastectomy aimed at minimizing unnecessary skin sacrifice of the breast when immediate breast reconstruction is planned. Preservation of breast skin unaffected by tumor provides an ideal color and texture match of the reconstructed breast with the opposite breast. The advantages of immediate breast reconstruction are its cost-effectiveness--only 1 anesthesia induction and hospitalization are required--and an improved cosmetic outcome that translates into a positive body image for the patient. Skin-sparing mastectomy with immediate reconstruction does not increase local or regional cancer recurrence or interfere with adjuvant radiation therapy or chemotherapy. The 2 techniques employed for creating a reconstructed breast mound are saline- or silicone-gel-filled implants and the use of autogenous tissue. PMID- 9746647 TI - Bridging the Gender Gap in HIV Diagnosis and Care. AB - Since its emergence in 1981, the human immunodeficiency virus (HIV) has been viewed by the Western world as a disease that primarily affected men. Yet in mid 1996, the World Health Organization (WHO) estimated that globally 42% of the 21 million adults living with HIV infection were women. They also reported that the proportion of infected women continues to grow in developed and underdeveloped countries. The clinical course and complications of HIV infection are similar in women and men, but women are frequently not diagnosed until they present with advanced HIV disease because they are not usually perceived to be at risk. Some manifestations of acquired immune deficiency syndrome (AIDS) are unique to women, and these clinical manifestations may be the first recognizable indications of AIDS in women. For example, gynecologic symptoms develop in more than 70% of HIV infected women, and these are often the presenting clinical manifestation. Although management principles for women are nearly identical to those of men, there are some unique aspects. The authors review specific manifestations of AIDS in women, discuss the issues of contraception and pregnancy, and address the psychosocial and economic issues contributing to the gaps that exist in the current level of care. PMID- 9746648 TI - Intrauterine Devices: Separating Fact From Fallacy. AB - IUDs currently available in the US provide safe and effective contraception. History has falsely led patients and clinicians alike to believe that IUDs are unsafe. For a woman in a long-standing, mutually monogamous relationship, no method of reversible contraception is more effective. The main risk associated with an IUD is infection; this is usually related to insertion, and the risk lasts for approximately 20 days. Currently available IUDs work by various mechanisms to prevent fertilization. The copper-containing IUD is also a highly effective form of emergency contraception. It is important for clinicians to re educate themselves and their patients about the importance of the IUD as an option for contraception. PMID- 9746649 TI - Advances in Therapy for Advanced Breast Cancer. AB - Breast cancer is a major public health problem in the US, with 46,000 women dying annually from the disease. However, innovative therapies are rapidly attempting to change the natural history of advanced disease. These therapies include such new drugs as taxanes and innovative modalities like gene therapy, immunotherapy, therapy with monoclonal antibodies, and higher doses of established agents in high-dose chemotherapy with stem cell support. Clinical trials are currently exploring all of these modalities. PMID- 9746650 TI - Intrauterine Exposure to Drugs and Alcohol: How Do the Children Fare? AB - The increasing use of drugs and alcohol by pregnant women has resulted in a population of children with unique neurodevelopmental behaviors. Of 223 children evaluated in a multidisciplinary child-development program in Portland, Oregon, 188 required long-term follow-up because of neurodevelopmental and/or behavioral problems; 153 had been polydrug-exposed in utero and 35 were born with fetal alcohol syndrome (FAS). Evaluations of the polydrug-exposed children revealed that 13% had severe speech and language disorders, 10% had significant cognitive deficits, and 81% had behavior problems after the age of 3 years. These deficits could not be attributed to social/environmental factors alone and probably reflect the effect of toxins on the developing brain. While polydrug-exposed children had learning problems related to impaired neurodevelopment and attention deficits, their IQs were normal. Among children with FAS, however, 67% had some degree of mental retardation. PMID- 9746651 TI - Treating CAD with Cardiac Surgery Combined with Complementary Therapy. AB - Coronary artery bypass graft (CABG) surgery to bypass blocks in the vascular pipeline can correct an acute problem, but it doesn't treat the underlying disease. This cardiac surgical team from a leading medical center explains why and how music therapy, aromatherapy, hypnosis, diet, and other modalities can be used to complement and extend the effect of the surgical repair. PMID- 9746652 TI - Can Antidepressants be Used to Tame Psychological Symptoms of PMS? AB - Up to 80% of women of reproductive age experience physical and behavioral changes premenstrually. However, these changes are predominantly normal and most often do not result in disabling distress or functional impairment. Still, 20% to 40% of women do experience premenstrual disorders. For 2% to 10% of women, the symptoms are severe enough to impair work and interpersonal relationships. Only in recent years has there been significant recognition and research of the problem. Diagnostic criteria are now evolving, and well-designed studies of treatment efficacy are providing greatly improved treatment information. No single treatment is effective for all women with premenstrual syndrome (PMS), although most women can be helped with careful diagnosis and a systematic approach to potential therapies. The overlap between PMS and atypical depression--in which emotional hypersensitivity, increased anxiety, irritability, and food cravings predominate--is particularly notable. Both disorders are believed to involve the serotonin system, which accumulating evidence shows to have a role in modulating mood and behavior. Results of various preliminary studies that examined markers and activity of the serotonin system are consistent with the hypothesis that abnormalities in central serotonergic activity may be involved in PMS. Clinical studies suggest that approximately 60% to 70% of women who have undergone a careful diagnostic evaluation of severe PMS report significant symptom reduction with the serotonergic antidepressants, although as yet there are no drugs with FDA approval for the indication of PMS. Continued research is essential to confirm and expand the pharmacologic treatments for PMS and to increase understanding of this complex disorder. PMID- 9746653 TI - HIV in Women: Recognizing the Signs. AB - The rates of gynecologic disease in HIV-positive women have been shown to be as high as 42%; given this high percentage, a routine GYN exam can be an invaluable opportunity for the detection of subtle signs of HIV infection. PMID- 9746654 TI - Prelabor Rupture of Membranes at Term: Induce or Wait? AB - Rupture of the fetal membranes prior to the onset of labor is a common event. With its occurrence, the risk is increased for serious maternal and fetal infections, dysfunctional labor, and the need for operative delivery. Fortunately, for most women with prelabor rupture of membranes (PROM) at term, labor begins spontaneously in the hours following membrane rupture. If spontaneous labor does not commence shortly after PROM at term, and the patient's cervix is favorable, labor induction should be initiated. Women with uncomplicated gestations who have an unfavorable cervical score may be managed expectantly, undergo immediate labor induction, or be observed for an arbitrary time period and then undergo labor induction. Recent clinical trials have shown that immediate labor induction for women with PROM at term and an unfavorable cervical score is preferable in the context of maternal and neonatal outcome, cost-effectiveness, and patient satisfaction. PMID- 9746655 TI - Ovarian Cancer Controversy: When and How To Use Available Screening Methods. AB - Ovarian cancer is the fourth most common cause of cancer death in American women. While the prognosis is excellent when the cancer is confined to the ovary, the majority of women are diagnosed after the tumor has spread to the upper abdomen, and their prognosis is poor. To diagnose the disease before it has spread, numerous studies have evaluated various methods of ovarian cancer screening. For example, such cytologic screens as cervical Papanicolaou smears and culdocentesis for cytologic analysis of cul-de-sac fluid have been studied, but they have been shown to be ineffective. Serum markers like CA-125 have proved useful in monitoring the progression of ovarian cancer after diagnosis, but the long list of other conditions that can elevate CA-125 hamper its usefulness in screening. Additional serum markers under current investigation include OVX-1, M-CSF, CA-15 3, CA-19-9, and TAG-72.3. Many investigators are studying imaging techniques such as transvaginal sonography (TVS). To distinguish between benign and malignant tumors seen in visualization studies, adjuncts such as color flow Doppler imaging to assess blood flow through a tumor, and tumor morphology indices that assign values to sonographic variables (eg, ovarian volume or cyst wall thickness) are also under study. The National Cancer Institute is currently performing a multicenter, randomized screening trial for ovarian cancer using the combination of serum CA-125 and TVS. However, until the results of this trial become available, the combined use of serum CA-125 and TVS cannot be recommended for ovarian cancer screening in the general population; these tests should be limited to clinical research settings and to those patients with a documented family history of ovarian cancer. PMID- 9746656 TI - Ovarian Cancer Controversies: Examining the Options After Primary Treatment. AB - Overall 5-year survival for patients with epithelial ovarian cancer drops from 88% for stages I and II, to 36% for stage III, and to 17% for stage IV. Over the past 15 years, mortality rates from ovarian cancer have fallen slightly for women under age 65 and have risen slightly for women older than 65. Since paclitaxel was approved for commercial use in 1992, we do not yet know how it will affect 5 year survival figures. The usefulness of primary adjuvant chemotherapy in women with early stage I or II ovarian cancer is unclear, but for women with advanced ovarian cancer, primary treatment should include 6 courses of platinum/paclitaxel chemotherapy, followed by second-look surgery to help women and their physicians decide on appropriate consolidation therapy. Potential consolidation treatments- including continuation of intravenous platinum and paclitaxel therapy, intraperitoneal cisplatin and/or paclitaxel therapy, topotecan, and high-dose chemotherapy with hematologic support--offer the promise of rendering primary therapy more effective. For follow-up after primary therapy, the benefits of routine CA-125 and imaging studies have not been established. In this clinician's practice, patients who have no evidence of disease are followed with office visits and physical examination every 6 months for 5 years; CA-125 levels or order imaging studies are not routine unless required by protocol. Women with symptomatic, recurrent disease may be treated with chemotherapy and, in some cases, surgery. Maintenance of quality of life and palliation of symptoms are crucial. PMID- 9746657 TI - Autoimmunity: The Female Connection. AB - The prevalence of autoimmune diseases in women may be the consequence of a bidirectional signaling network between hormones and the immune system that regulates female reproductive life. Two prototypical autoimmune diseases, rheumatoid arthritis and systemic lupus erythematosus, arise from 2 different immune responses that generate mutually exclusive signals in response to different inflammatory triggers. Certain estrogens may ameliorate the rheumatoid arthritis-like TH1 response while exacerbating the lupus-like TH2 response. Studies of sex hormone metabolism in lupus patients reveal increased 16 hydroxylation of estrone in some patients and decreased levels of androgens as a result of increased oxidation at C17. These occurrences result in low serum levels of dehydroepiandrosterone (DHEA). Both the increase of 16-hydroxylation of estrone and the depletion of DHEA have immune effects that would tend to exacerbate a lupus-like TH2 response. This theoretical framework provides a rationale for ongoing initial clinical trials of exogenous hormones in autoimmune diseases. PMID- 9746659 TI - Gynecology Case Challenge - Suspicious Cervical Structure. AB - A 23-year-old multiparous woman is referred for an asymptomatic suspicious cervical lesion. What is your diagnosis and treatment plan? PMID- 9746658 TI - The High Risk of CHD for Women: Understanding Why Prevention Is Crucial. AB - Coronary heart disease (CHD) is the leading cause of death and disability among adult women in the US. A postmenopausal woman has a 31% lifetime mortality risk from CHD, in contrast to a 2.8% mortality risk from hip fracture and a comparable risk for breast cancer. As many as 1 in 8 women aged 45 to 54 years has clinical evidence of CHD, with the prevalence increasing to 1 in 3 women older than 65 years of age. Once women develop clinical evidence of CHD, their outcomes are less favorable than those for their male counterparts. Data from the Myocardial Infarction Triage and Intervention Registry reveal a hospital mortality rate for acute myocardial infarction of 16% for women, in contrast to 11% for men; 1-year mortality also was greater in women. Among the risk factors that heighten the risk of CHD in women are diabetes, which negates the gender-protective effect, even for premenopausal women; cigarette smoking, which is on the rise in young women; hypertension, which is more prevalent in women than men after 65 years of age; LDL cholesterol levels that begin to rise in women after menopause and exceed levels in men at elderly age; and triglyceride levels that are higher in men than in women at all ages. Oral contraceptives, now prescribed at lower dose, are no longer considered a serious CHD risk factor since they have fewer adverse effects on glucose tolerance, insulin resistance, and lipoprotein levels. Because of the high prevalence of traditional coronary risk factors among US women, preventive strategies are likely to effect substantial benefit. Risk attributes unique to women are topics of active research. PMID- 9746660 TI - Rating the Chemotherapy Options for Advanced Uterine Adenocarcinoma. AB - During 1996 it is estimated that 34,000 women will develop endometrial adenocarcinoma, and 5900 will succumb to the disease. However, in nearly 75% of patients the diagnosis is made early, when the disease is confined to the uterus and easily treated with surgery. Approximately 25% of patients will have operative findings that make them appropriate candidates for postoperative adjuvant radiotherapy. For the minority of patients who present with metastatic or recurrent disease not amenable to local control measures or oral hormonal therapy, chemotherapy can be administered. The following agents are known to have activity against endometrial adenocarcinoma: doxorubicin, cisplatin, carboplatin, ifosfamide, and paclitaxel. Cyclophosphamide, 5-fluorouracil, and hexamethylmelamine are drugs that have shown promise, but data are as yet too sparse to draw definite conclusions about their effectiveness against endometrial adenocarcinoma. The 2 most commonly employed hormones used to suppress tumor growth have been megestrol acetate and medroxyprogesterone acetate. Based on the responses reported with hormonal therapy alone, some investigators have combined multiagent cytotoxic therapy with hormones. The highest response rates utilizing chemohormonal regimens have been reported when the cytotoxic regimen included doxorubicin and cisplatin. We review the response rates demonstrated in single- and multi-institutional trials of these agents when used alone or in combination to treat patients with advanced or recurrent metastatic endometrial adenocarcinoma. PMID- 9746661 TI - Ugly Risks of Beauty Routines. AB - Beauty products and practices may put women at risk for infection. Adverse events range in seriousness from the superficial to the systemic--from conjunctivitis caused by contaminated eye makeup, to fungal infections picked up from the locker room or manicurist, to more troubling soft-tissue infections caused by unsanitary body piercing or tattooing, practices that can also transmit hepatitis B and C. PMID- 9746662 TI - How HRT Alters the Lipid Profile in Women with Diabetes. AB - Although postmenopausal estrogen replacement is recommended to prevent disease and prolong life, little data are available regarding hormone replacement therapy (HRT; estrogen or estrogen plus progestin) in various high-risk subpopulations of women. Our study examined lipid levels, with and without HRT, in a cross-section of 694 postmenopausal women with diabetes and 5321 postmenopausal women without diabetes. Among the diabetic women, 70 were currently using HRT, 482 had never used HRT, and 142 had used HRT at some point after menopause but were not taking hormones at the time of the study. Among the nondiabetic women, 1147 were currently using HRT, 3210 had never used HRT, and 964 had formerly used HRT. We found that diabetic women appear to respond somewhat differently than their nondiabetic counterparts to HRT. Estrogens were associated with proportionately lower increases in HDL as compared with the increases produced in nondiabetic women--diabetic women currently taking estrogen had 6% higher HDL levels than diabetic women who never used estrogen, and nondiabetic women currently taking hormones had 17% higher HDL levels than nondiabetic subjects who were never on HRT (P=0.02). Further, there were proportional differences in triglyceride levels--diabetic women currently on HRT had 25% higher triglyceride levels than diabetic women who never used hormone therapy, and among nondiabetic women on HRT, there was a 14% increase in triglycerides compared with their counterparts who were never on HRT (P=0.08). LDL cholesterol appeared to respond similarly to HRT in diabetic and nondiabetic women. Diabetic women appear to have a blunted response to the HDL-raising effect of HRT and an exaggerated hypertriglyceridemic response. This may alter the cardiovascular benefits from postmenopausal HRT and increase the risk of acute pancreatitis from hypertriglyceridemia. The decision to use HRT should be individualized, and diabetic women who receive HRT should have their triglycerides carefully monitored at 1 month and then at every 3 months after starting therapy. PMID- 9746663 TI - What Can Be Done to Prevent Coronary Heart Disease in Women? AB - Despite the decrease in coronary heart disease (CHD) mortality in the US in the past 30 years, CHD is the leading cause of death in men and women. Cardiovascular disease, including CHD, kills nearly 500,000 American women each year. In women, the development of CHD can be delayed by an average of 10 years compared with men, and, on average, women can experience a first myocardial infarction 20 years later than men. While CHD prevalence rates are similar in black men and white men, heart disease is not color-blind in women. Black women generally have a higher prevalence of CHD risk factors and a higher death rate at a younger age than white women. A strong family history of early onset of heart disease, increasing age, and race are unalterable factors that raise the risk of CHD. The major factors that can be modified include cigarette smoking, hypertension, diabetes mellitus, physical inactivity, obesity, dyslipidemia, estrogen level changes after menopause, and psychosocial stressors. CHD is a multifactorial process; the hazard posed by one particular risk factor is significantly influenced by other risk factors that are present, and no individual risk factor is essential or sufficient to cause CHD. PMID- 9746664 TI - Understanding How Neural Tube Defects Occur--And Can Be Prevented. AB - Neural tube defects occur in 1.3 to 2.0 fetuses per 1000 live births, and are second only to cardiac malformations as the most frequent congenital anomaly in the US. The genetic and environmental influences on neural tube defect formation are currently being investigated. It is now thought that neural tube closure occurs in 5 separate sites, with each site possibly controlled by separate genes and subject to different external influences. Evidence suggests that one basic genetic defect involves homocysteine metabolism. Folic acid supplements before conception reduce both occurrence and recurrence risks. Prenatal screening and diagnosis are available, although management options after diagnosis are limited. Dietary alterations and adjustment of medical therapy prior to conception will reduce neural tube defect occurrence. PMID- 9746665 TI - Assessing and Managing Urinary Incontinence in Primary Care. AB - Incontinence generally results from a problem either with the urethra or with the bladder. The urethra can permit urine to leak when it moves incessantly due to poor support or when it closes poorly, as can occur with a neurologic disorder. In the bladder, hyperactivity due to involuntary contractions, or low compliance, can lead to urinary incontinence. A thorough history and physical examination guide management. The history should elicit information that can allow the clinician to assess how severe the problem is (eg, number of pads used per day, frequency of leakage) and factors that may cause or influence the problem, such as medications used, previous surgical and obstetric history, and urologic history including prior therapy for incontinence. Information about when leakage is at its worst is useful. Leakage that worsens in winter is typically associated with detrusor instability. Leakage that worsens at night can indicate a problem with bladder compliance. Incontinence that started after the onset of an antihypertensive or antipsychotic medication may be due to alpha-receptor antagonist effects of drugs such as prazosin or chlorpromazine. Difficulty emptying the bladder may be associated with medications that block cholinergic and calcium-channel activity, such as sedatives, antidepressants, antispasmodics, antiemetics, antipsychotics, antiarrhythmics, and anticonvulsants. Mild incontinence can be managed conservatively in a primary care setting, with pelvic floor exercise, behavior therapy, or anticholinergic therapy. Patients with severe incontinence or an unclear etiology of incontinence should be referred to a specialist for urodynamic testing. PMID- 9746666 TI - Contemporary Concepts in Managing Menorrhagia. AB - Menorrhagia--technically defined as menses lasting longer than 7 days or a blood loss volume in excess of 60 to 80 mL--is one of the most common gynecologic complaints. It has been reported that 15% to 20% of otherwise healthy women experience debilitating menorrhagia. In the past, definitive treatment for abnormal uterine bleeding has been either abdominal or vaginal hysterectomy. Alternatives to hysterectomy are now stressed in light of the fact that nearly 50% of uterine specimens obtained during hysterectomies for menorrhagia are disease-free on pathologic examination. Etiologies are generally either endocrinologic or organic. Among the organic causes are coagulation disorders; organ dysfunction, such as liver or renal disease; endometrial hyperplasia; infection; iatrogenic causes, such as chemotherapy, anticoagulants, steroid therapy, and use of IUD; and anatomic causes, which include uterine leiomyoma, endometrial polyps, and pregnancy. Besides the history and physical exam, useful lab tests include CBC, serum pregnancy test, cervical specimens for gonorrhea and chlamydia, Pap smear, thyroid function tests, serum transaminases, luteinizing hormone, follicle-stimulating hormone, estradiol, prothrombin time, activated partial thromboplastin time, bleeding time, serum prolactin, quantitative beta human chorionic gonadotropin, blood urea nitrogen, serum creatinine, and adrenal function tests. Since many lesions are missed in office sampling or dilation and curettage, imaging studies, including ultrasound and hysteroscopy, can be useful in diagnosing the cause of menorrhagia. Medical treatments include drugs such as NSAIDS, progestins, oral contraceptives, and gonadotropin-releasing hormone agonists. Surgical modalities range from the relatively simple, such as D & C, to major surgical procedures such as hysterectomy. Laser ablation and thermal balloon ablation are promising new procedures. PMID- 9746667 TI - Universal Screening for HIV in Pregnant Women? AB - The fact that zidovudine therapy can prevent perinatal transmission of HIV infection strongly supports the need to screen for HIV during pregnancy. Controversy continues to revolve around which testing strategy offers the benefits of zidovudine to the greatest number of infants while still allowing the mother some degree of autonomy in making her own health care decisions. Screening based on patient-reported risk factors has repeatedly been shown to exclude many seropositive women. Voluntary testing has had inconsistent results, and mandatory testing may discourage women from getting the prenatal care they need. Despite the controversy, HIV testing and education should be recommended to all pregnant women. Counseling should be geared to the educational level of the patient, and risk-factor reduction is an important component of counseling. Health care providers must be prepared to address the social as well as the medical ramifications of a diagnosis of HIV infection for both the mother and the unborn child. PMID- 9746668 TI - Managing Back Pain During Pregnancy. AB - The incidence of low-back pain during pregnancy is thought to be about 50%. It occurs most commonly after the sixth month and can last until the sixth month postpartum. The major predictors are back pain prior to pregnancy and multiparity. Several biomechanical and physiologic changes during pregnancy contribute to back pain. As the woman's abdominal muscles are stretched and tone is diminished, they lose their ability to contribute to neutral posture. During pregnancy, production of the hormone relaxin increases ten-fold. The hormone creates joint laxity, which not only allows the pelvis to accommodate the enlarging uterus, but also weakens the ability of static supports in the lumbar spine to withstand shearing forces. In the pelvis, joint laxity is most prominent in the symphysis pubis and the sacroiliac joints. On physical exam, neither lumbar nor sacroiliac back pain is generally associated with any evidence of neurologic deficit or hip pathology. In cases of lumbar back pain, the physical exam will be most consistent with discogenic pain and/or facet element pain. Therefore, a pregnant woman's pain may be most pronounced on flexion and standing. Among the tests that can be used to evaluate lower-back pain are the posterior pelvic provocation test; ventral gapping test; dorsal gapping test; sacroiliac joint fixation test; Patrick's test, or FABERE's maneuver (flexion, abduction, external rotation, and extension); and Derbolowski's test. The most common types of back pain during pregnancy are lumbar pain, sacroiliac pain, and nocturnal back pain. PMID- 9746669 TI - Rett Syndrome: Meeting the Challenge of This Gender-Specific Neurodevelopmental Disorder. AB - Rett syndrome (RS) is an incurable neurological disorder that occurs in females. Although the biological basis is unknown, there is substantial evidence suggesting a genetic basis. RS is characterized by an initial period of apparently normal psychomotor development followed by loss of communication skills and purposeful hand movement. Then, hand stereotypies, gait dyspraxia, and deceleration of head growth become apparent. Other problems include growth failure and epilepsy. There is no biological marker for RS; the diagnosis is based on well-delineated clinical criteria. The prevalence of RS is 1:23,000 live female births. Survival to 30-40 years or beyond is the rule rather than the exception. Treatment is both palliative and supportive. A vigorous approach to all aspects of care, including educational, medical, and psychosocial issues, is recommended. PMID- 9746670 TI - Pharmacotherapy of Bulimia Nervosa. AB - This review article summarizes the progress that has been made over the past 14 years in the drug treatment of bulimia nervosa, an eating disorder characterized by recurrent binge eating and purging that has continued for 3 months with a regularity of at least twice a week. This recently described eating disorder has been the subject of intensive research scrutiny with regard to both pharmacotherapy and psychotherapy, and results indicate a role for both types of treatment. The linkage of bulimia to major depressive illness is reflected in the overlap that is seen in neurotransmitter dysregulations--including similarities in serotonin disturbances--as well as in shared symptomatology, longitudinal risk and comorbidity, and prominent history of affective illness in the families of bulimic probands. The observed relationship between these disorders acted as a catalyst for the early pharmacotherapy studies documenting the use of antidepressant medications in the treatment of bulimia. Among the drugs documented to reduce bulimia are tricyclic antidepressants, monoamine oxidase inhibitors, and selective serotonin reuptake inhibitors. The sequencing and integration of pharmacotherapy and psychotherapy require further study, but current research has yielded a variety of therapeutic options for the clinician to reduce both the disturbed eating behavior and the dysfunctional eating attitudes of patients with bulimia nervosa. PMID- 9746671 TI - Calcium Metabolism in Preeclampsia: Supplementation May Help. AB - Calcium homeostasis is an important aspect of maternal and fetal physiology during gestation, since fetal bone mineralization requires adaptive adjustments in maternal calcium regulation. In the third trimester, for example, calcium is deposited in the fetal skeleton at the rate of 200mg/day. In addition, women double their urinary excretion of calcium in the third trimester. Recent studies have implicated alterations in calcium metabolism in the pathogenesis of hypertension during pregnancy. Deficiencies in calcium intake have been linked to preeclampsia/eclampsia. Further, hypocalciuria and deviations in levels of 1,25 dihydroxyvitamin D and parathyroid hormone have been shown in women with preeclampsia. In one study, calcium supplementation was associated with an approximately 50% decrease in the risk of all types of pregnancy-induced hypertension. The risk of preeclampsia was between 45% and 74% lower for women who received calcium supplementation. At present, several trials of calcium supplementation in pregnancy are in progress. One, an NIH-sponsored study that will enroll more than 4000 patients, is nearing completion. PMID- 9746672 TI - Treatment Decisions in the Management of Menorrhagia. AB - Menorrhagia--menstrual periods lasting longer than 7 days and totaling blood losses greater than 80mL--affects 9%-14% of otherwise healthy women, and it can signal cancer, an endocrinologic disorder, or gynecologic disease. Blood loss can be high enough to result in anemia, fatigue, and syncope. Most often, abnormal uterine bleeding such as menorrhagia involves a disruption in the hypothalamic pituitary axis, the ovary, and/or the uterus. Other identified causes include medications (especially psychotropics) that cross the blood-brain barrier; chronic diseases such as cancer, diabetes, and liver and kidney dysfunction; endocrine disorders, perimenopausal anovulation, polycystic ovary disease, pituitary tumors, and abnormal estrogen cycling caused by morbid obesity; and anatomic abnormalities of the uterus. Routine tests include hematocrit or hemoglobin to detect and evaluate anemia, thyroid stimulating hormone (TSH) level to evaluate thyroid function as a possible cause, and a pregnancy test to rule out an incomplete, spontaneous abortion as a cause. A Pap test is recommended to screen for dysplasia that can suggest a gynecologic cancer cause. Additional screening for endocrine disorders that may be causing menorrhagia include tests of thyroid, liver, and kidney function, and tests of follicle stimulating hormone (FSH), prolactin, and cortisol levels. Treatment can be medical or surgical. Medical treatment includes prostaglandin inhibitors, specifically nonsteroidal antiinflammatory drugs (NSAIDs), and hormonal therapy with estrogen, progesterone, gonadotropin-releasing hormone agonists, or oral contraceptives such as medroxyprogesterone (Depo-Provera). Surgical treatment includes hysteroscopic endometrial ablation by physical agents, laser electrodiathermy, and "roller ball," or surgical, resection. Hysterectomy is the treatment of last resort. PMID- 9746674 TI - Gynecology Case Challenge: Uterine Prolapse? AB - Five weeks after a multiparous perimenopausal woman first noticed something "hanging out" of her vagina, she was referred for gynecologic evaluation. Looking through the speculum, what's your diagnosis? PMID- 9746673 TI - Impact of the Antiphospholipid Syndrome: A Critical Coagulation Disorder in Women. AB - Antiphospholipid (aPL) syndrome, or APS,--a cluster of conditions that includes arterial or venous thromboses and thrombocytopenia, as well as recurrent fetal loss associated with elevation of aPL antibody--has been reported to occur 2-5 times more frequently in women than men. Strong familial associations lead to the suspicion that aPL positivity, estimated to be present in 2% of the population, is a heritable trait in some cases. Currently, 2 major categories of the illness are recognized--primary and secondary. Secondary APS may be associated with autoimmune disease, malignancy, infectious disease, or drug-induced states. Two assays, one for lupus anticoagulant antibodies and the other for anticardiolipin (aCL) antibodies, are recognized to be the gold standards for serologic diagnosis of the disease. Despite extensive attempts at international standardization of aCL test results, no consensus exists for a value beyond which the test is considered positive. Interestingly, a "dose-effect" relationship for aCL antibody titers has been noted--higher titers of the antibody correlate with increased numbers of thrombotic events. An experimental assay for antibody against beta 2 glycoprotein 1 (beta-2-GP1), a phospholipid-binding protein, may become the most important assay for aPL. Skin findings in APS include livedo reticularis, ulceration, gangrene, or purpura, and, when present, may be the key to diagnosis of this sometimes insidious syndrome. Anticoagulation, usually with warfarin, is the mainstay of therapy, although steroids, immunosuppressive agents, hydroxychloroquine sulfate, and plasmapheresis may all be beneficial adjunctive therapy. PMID- 9746675 TI - Breast Cancer Screening For Women Ages 40-49--NIH Consensus Statement. AB - For 2 days, January 21 and 22, a panel of experts representing specialties in women's health, oncology, radiology, epidemiology, and public health listened to presentations by researchers and to public discussion and debate designed to reach a consensus on 5 questions: (1) Is there a reduction in breast cancer mortality due to screening women ages 40-49 with mammography, with or without physical examination? (2) What are the risks associated with screening women ages 40-49 with mammography and with physical examination? (3) Are there other benefits? If so, what and how do they change with age? (4) What is known about how the benefits and risks of breast cancer screening differ based on known risk factors for breast cancer? (5) What are the directions for future research? On January 23, the panel issued a consensus statement concluding that, while mammography has been shown to be clearly effective in reducing cancer mortality in women ages 50-69, there is no difference in breast cancer death within 7 years between 40- to 49-year-old women assigned to receive or not receive screening. For women in their 40s, the potential benefits of earlier diagnosis and breast conserving therapy must be weighed against the potential risks of mammograms, including discomfort, inconvenience, radiation, and false reassurance given to women with false-negative screens. At the present time, the available data do not warrant a single recommendation for mammography for all women in their 40s. Each woman should decide for herself whether to undergo mammography. PMID- 9746676 TI - Practical Guide to Diagnosing and Treating Vaginitis. AB - Bacterial vaginosis (BV), candidiasis, and trichomoniasis account for more than 90% of vaginal infections. BV typically is associated with a decrease in commensal, protective lactobacilli and a proliferation of other flora. Mobiluncus is pathognomonic but found in only 20% of cases. Presence of 3 of 4 criteria indicates BV: a homogenous noninflammatory discharge (not many WBCs); pH >4.5; clue cells (bacteria attached to borders of epithelial cells, > 20 % of epithelial cells); and a positive whiff test. New intravaginal BV preparations cause less-adverse systemic effects than oral regimens. Trichomonas vaginalis, a protozoan, appears to be sexually transmitted and causes up to 25% of vaginitis cases. Diagnosis is made by observation of a foul, frothy discharge; pH >4.5 (present in 70% of cases); punctate cervical microhemorrhages (25% of cases); and motile trichomonads on wet mount (50%-75% of cases). Recommended treatment is a single 2g dose of oral metronidazole. Treatment failure is usually due to nontreatment of the male partner. Candidiasis typically presents as a thick, "curdled" white discharge or vulvar pruritus, with a hyperemic vagina and an erythematous and/or excoriated vulva. Vaginal pH is usually in the normal range of 3.8-4.2 in uncomplicated candidiasis. Microscopic examination of the discharge reveals hyphae or budding yeast in 50%-70% of cases. While the most common offender is Candida albicans, Candida tropicalis and Candida glabrata have become increasingly prevalent. Approximately 15% of C albicans organisms are resistant to clotrimazole and miconazole. Recurrent infections may be treated with fluconazole 150mg weekly for up to 12 consecutive weeks. PMID- 9746677 TI - Gender Differences in Depression. AB - Beyond the repeatedly confirmed finding that women diagnosed with mood disorders greatly outnumber men lies a widely varying set of hypotheses that attempt to explain the suspected causes, incidence, symptoms, and comorbidities from various perspectives. Several complex factors, however, have impeded attempts to study why women are so vulnerable to depression. This article examines the problems associated with studying affective disorders in women and reviews the current hypothetical constructs of the etiology and pathophysiology of depression and their potential relevance to the disproportionate number of women with unipolar depression. The association of depression to biological stages of a woman's life and the differences between the biology of men and women are described, and the potential social, psychological, and environmental factors that might particularly promote the development of depression in women are discussed. PMID- 9746678 TI - Inflammatory Bowel Disease--A Complicating Factor in Gynecologic Disorders? AB - Gynecologic disorders occur commonly in women with Crohn's disease and ulcerative colitis. Frequently, these women also suffer menstrual disorders with gastrointestinal symptoms that overlap with those related to inflammatory bowel disease (IBD). Knowledge of the range of gynecologic problems--for example, dysfunctional uterine bleeding, fistula or abscess of the perineum or vagina, dyspareunia, subfertility possibly due to tubal blockage, and ovarian dysfunction related to bowel disease--that have been associated with IBD will assist practitioners in treating these women. Prostaglandins, released by the endometrium at menstruation, cause contraction of uterine smooth muscle, resulting in the cramping pain of dysmenorrhea. Prostaglandins also are an important component of the inflammatory process in active IBD; by increasing contractility of GI smooth muscle, they are associated with diarrhea and abdominal pain. Menstrual pain and menses-related GI symptoms may be difficult to distinguish from symptoms related to IBD. Endometriosis may present with symptoms similar to an acute episode of IBD. Mucosal changes in the bowel can occur in association with endometriosis, and can be confused with the histologic features of IBD. The distinction is important. For example, while nonsteroidal anti inflammatory drugs may relieve symptoms of dysmenorrhea, they often are contraindicated in IBD. To provide optimal evaluation and treatment, all health care professionals who treat women with IBD should be aware of the spectrum of gynecologic conditions that may be encountered. PMID- 9746679 TI - Examining the Gender Bias in Evaluating Coronary Disease in Women. AB - Gender discrepancies have developed in the evaluation of coronary heart disease (CHD), arising from such early myths as "CHD is a man's disease." The challenge is to make sure that the noninvasive testing for CHD in women is sensitive and specific enough to lead to the correct treatment. Coronary angiography, the gold standard for CHD diagnosis, must be interpreted along with functional information. The standard noninvasive test--stress electrocardiograph (ECG)--is associated with up to 40% false-positive S-T segment depressions in women, versus fewer than 10% in men. The predictive value of exercise stress testing in women is particularly poor. In one study, stress ECG had a specificity of 61%, a sensitivity of 68%, a positive predictive value of 0.61, and a negative predictive value of 0.68. Stress echocardiography can have high sensitivity (86%) and specificity (86%), but often examiners stop the test before detecting less severe areas of damage. Also, acquiring adequate images is difficult in women with breast implants or large breasts. Nuclear perfusion imaging with thallium 201 has shown a sensitivity of 84% to 90% and a specificity of 75% to 87% in women, but the diagnostic accuracy can be reduced in patients who are obese or have large breasts. A higher-energy radiotracer, technetium-99m (Tc-99m) sestamibi, has been introduced. In one study, the sensitivity of the 2 agents was similar (85% to 90%), while the specificity of Tc-99m was higher (84% to 94%) than that of thallium-201 (71%). PMID- 9746680 TI - Diagnosing and Treating the Predominantly Female Problems of Systemic Autoimmune Diseases. AB - Autoimmune diseases with rheumatic manifestations are predominantly diseases of women. Any woman with new onset arthralgia or arthritis should have a thorough history and physical to rule out autoimmune connective-tissue disease. Screening serologic tests, however, are not necessarily recommended because of high false positive rates. Serologic tests are most useful in confirming or ruling out the diagnosis; for example, systemic lupus erythematosus (SLE) is rarely present when antinuclear antibodies (ANAs) are absent; the absence of rheumatoid factor will not rule out rheumatoid arthritis (RA), but its presence confirms it. Most autoantibodies (RF [rheumatoid factor], ANA, ENA [extractable nuclear antigen], anti-Jo-1, etc) do not vary with disease flare-ups and remissions. Plain radiographs of the joints are not useful except as a baseline and in detecting erosion at end of long bones associated with RA. Polymyalgia rheumatica and RA, with an incidence of 1 in 2000 to 3000, are the most common autoimmune disorders. Other autoimmune diseases such as SLE, vasculitis, polymyositis, and dermatomyositis are seen infrequently in general practice. Pregnancy, menopause, or breast implantation may affect disease prognosis and treatment. For example, in pregnancy, RA symptoms generally improve, whereas those of SLE may worsen; both diseases may flare postpartum. Oral contraceptives have been associated with an increase in disease flare-ups, but there is little or no evidence that estrogen in the dose level used for replacement is harmful to SLE patients. Although relatively rare, autoimmune diseases can be devastating to the patient if not promptly recognized and properly treated. PMID- 9746681 TI - BRCA 1 and 2--A Genetic Link to Familial Breast and Ovarian Cancer. AB - Female carriers of germline BRCA1 mutations have a lifetime risk of breast cancer exceeding 80% and of ovarian cancer approaching 60%. The cumulative lifetime risk of developing either breast or ovarian cancer, therefore, approaches 100%. Carriers of BRCA2 mutants have a similar risk of breast cancer and a more moderately increased risk of ovarian cancer. BRCA1 and BRCA2, located on the long arms of chromosomes 17 and 13, respectively, are thought to be tumor suppressor genes, inhibiting tumor development when functioning normally. Both are large genes, distributed over approximately 100,000 base pairs of genomic DNA, encoding large negatively charged proteins. Inactivating mutations identified to date are distributed throughout both genes, with an increased frequency of two distinct BRCA1 mutations and one BRCA2 mutation in individuals of Ashkenazi Jewish descent. Given the high lifetime penetrance of germline BRCA1 and BRCA2 mutations and the early age of onset in many carriers, it may seem prudent to carry out regular mammography on carriers from a young age. The benefits or risks of such screening, however, have yet to be demonstrated. Preventative measures, including prophylactic mastectomy, oophorectomy, and chemoprevention, still require assessment within research protocols. PMID- 9746682 TI - Gynecologic Case Challenge - Asymptomatic Cervical.? AB - A 42-year-old multiparous woman is referred to you for evaluation of an asymptomatic cervical lesion. What is your diagnosis and treatment plan? PMID- 9746683 TI - Examining PTSD as a Complication of Infertility. AB - Posttraumatic stress disorder (PTSD) is a psychiatric disorder that may develop following exposure to threatened or actual injury or death. While commonly associated with war or natural disaster, symptoms of PTSD have been described in patients who are undergoing or who have completed infertility treatment or high risk pregnancies. Three case studies of patients who developed PTSD following such pregnancies are discussed, demonstrating the variety of symptoms and presentations of these patients. The clinician must be vigilant in monitoring infertility patients with PTSD. These women, as the result of infertility, may be at increased risk of developing PTSD, one of the recognized postpartum psychiatric disorders. It is important to distinguish PTSD from postpartum depression, because treatment guidelines vary. PMID- 9746684 TI - Cervical Neoplasia and Cigarette Smoking: Are They Linked? AB - The presence of tobacco-specific carcinogens in the cervical mucus of smokers and their effect on the local immune system strongly suggest that smoking has an etiologic role in the development of cervical neoplasia. However, it remains unclear whether smoking can affect the initiation of high-grade cervical neoplasia independently from human papillomavirus (HPV) infection. Studies that control for HPV infection may not entirely resolve the issue of the role of smoking in cervical neoplasia. Cigarette smoking may be causative through its effect on oncogenic HPV infection or by altering the immune response system. This article reviews the currently available data assessing the relationship between cigarette smoking and cervical neoplasia. PMID- 9746685 TI - Understanding, Recognizing, and Treating Rett Syndrome. AB - Formerly thought to be a neurodegenerative disease, Rett syndrome (RS) is a neurodevelopmental arrest of the brain that almost exclusively affects females and occurs in a variety of racial and ethnic groups worldwide. RS begins in late infancy and is characterized by autistic and dementia-like behavior, ataxia, and purposeless hand movements. Its cause and mode of transmission are unknown in over 90% of cases; however, there is strong and convincing evidence that genetic factors play a major role. The reported incidence varies, but in the US, as many as one quarter to one third of female children in mental wards/institutions may be affected. RS has been mistaken for numerous other conditions, including autism, cerebral palsy, and mental retardation, but the clinical picture is unique: No other condition has a period of rapid deterioration followed by apparent stabilization or even improvement in autistic features, eye contact, seizure activity, and hand stereotypies. The diagnosis is supported by deceleration of head growth, evidence of neurologic regression with associated neurologic signs, and purposeless hand stereotypies, with a clinical history of developmental regression. The differential diagnosis often involves ruling out syndromes with similar signs of neurodevelopmental arrest--for example, meningitis or encephalitis; chromosomal disorders such as Angelman's syndrome and Prader-Willi syndrome; metabolic disorders such as ornithine carbamoyltransferase deficiency; disorders of organic acids and amino acids; neurovisceral storage diseases; mitochondrial cytopathy; and Batten disease, or infantile neuronal ceroid lipofuscinosis. Management encompasses a comprehensive medical, therapeutic, educational, and psychosocial approach, best provided through a team in collaboration with the community agencies that serve families and children with special needs. PMID- 9746686 TI - Examining Women's Health: 1996-1997. AB - Heart disease, lung cancer, and HIV infection are among the diseases previously thought to be primarily men's health problems that have been documented in recent years to be serious health problems for women. Researchers have reported that women with heart disease have poorer outcomes and receive less intensive therapy than men. Clinicians and consumers are just beginning to realize that cardiac disease is the #1 cause of death in women -- outpacing breast cancer. In the breast cancer arena, the impact of such genetic links as BRCA1 and BRCA2 is still unclear, as is the issue of screening mammograms for women under the age of 50. Other top issues in women's health include efforts to ban "drive through" deliveries and early postmastectomy discharge, calculation of the high price of prematurity, changes in Pap screening techniques, and continuing efforts to understand the effects of estrogen. This editorial examines the key issues and trends in women's health reported and debated in 1996. PMID- 9746687 TI - Vaginectomy: Profile of Success in Treating Vaginal Prolapse. AB - Treatment of procidentia and vaginal inversion in older women either with pessaries or surgery commonly brings poor results. Women are unable to retain the pessary; they develop vaginitis and vaginal ulcerations; and surgical "correction" fails due to age-induced genital atrophy or previous obstetrical trauma. We performed a retrospective chart review to assess results of our own technique of vaginectomy/hysterectomy and pelvic floor closure for vaginal vault prolapse and procidentia in 26 aged sexually inactive women seen in our practice. The women ranged from 63 to 83 years of age and had borne 0 to 9 children. Where possible, an estrogen-containing medication was introduced into the vagina preoperatively to stimulate thickening of the vaginal mucosa. A standard Heaney or Doderlein vaginal hysterectomy was performed. Operative time averaged 100 minutes, blood loss averaged 278mL, and 5 patients required a blood transfusion. All patients were discharged in good condition after an average stay of 4.67 days, although 9 of the 24 patients had complications. Since body-cavity invasion was minimal, postoperative care was simple, consisting of hydration with intravenous fluids, urine drainage utilizing an indwelling catheter, (while preventing bladder distension), early ambulation, and prophylaxis against infection and thromboembolism. PMID- 9746688 TI - NIH Consensus Statement on Breast Cancer Screening for Women in Their 40s: How Will It Affect Patient Care? AB - It is generally accepted that breast cancer screening mammograms in women 50 years of age and older has saved lives. It also is generally accepted that the incidence of breast cancer in women younger than 40 is too small to warrant subjecting young women to the risks associated with mammograms. But whether women in the transition years from 40 to 49 should have routine screening mammograms has been debated for 2 decades. On January 23, the NIH Consensus Statement on Screening Mammograms for Women Ages 40 to 49 lit a powder keg when, after 2-and-a half days of hearings and study, it announced "The data do not support a universal recommendation that all women in their 40s should undergo screening mammography." Many experts have challenged the conclusion and advocated screening mammograms every 1 to 2 years starting at age 40. To sort out the impact of the NIH statement and determine where clinicians and consumers stand, Medscape launched its first on-line survey on January 30. Most Medscape responders agree that screening mammograms every 1 to 2 years should begin at age 40 and fear that third-party health care payers will use the conclusion of the NIH consensus panel to deny reimbursement for screening mammograms in women younger than 50 years of age. PMID- 9746689 TI - Breast Cancer Imaging: Can Tc-99m Sestamibi Scintimammography Fit In? AB - Mammography and physical examination are currently the most frequently used screening tests for breast cancer. Considering the 85% sensitivity associated with combined mammography and physical examination and a low positive predictive value of 20% to 30% for the diagnosis of breast carcinoma, there is a critical need for a more accurate noninvasive imaging test to improve the sensitivity and specificity of mammography. This study evaluates the role of Tc-99m sestamibi scintimammography as a complementary procedure to conventional mammography for the detection of breast carcinoma. A sample of 157 women (mean age 47.9 years +/- 10.2 years) with 164 lesions appropriate for histologic and cytologic analysis on the basis of suspicious findings on a mammogram and/or physical examination underwent scintimammography. Subsequently, excisional biopsy and/or fine-needle aspiration were performed. There were 52 primary cancers (8 different histopathologic types) and 112 benign breast lesions (6 different histopathologic types). The sensitivity of Tc-99m sestamibi scintimammography for detecting primary breast cancer was 92.3%, and its specificity was 87.5%. Percent-positive and -negative predictive values associated with Tc-99m sestamibi scintimammography in this cohort were 77.4% and 96.0%, respectively. PMID- 9746690 TI - Facial Hair on a Woman: Diagnosing and Treating a Pathological Twist on a Common Problem. AB - A 46-year-old woman who had had occasional coarse, dark hairs on her chin and chest since she was 17 years old presented with rapidly progressive hirsutism and new onset of virilization--(eg, for the first time in her life, she had coarse, dark hair on her back and balding in the temporal and occipital areas of her scalp). A thorough evaluation, including laboratory tests, several imaging studies, and ovarian and adrenal vein catheterization, revealed a small ovarian hilus cell tumor that was successfully removed by a laparoscopic approach. During the 30 months after the testosterone-producing ovarian tumor had been diagnosed and the woman's ovary had been removed, her hirsutism progressed no further, hair began to grow back in the temporal and occipital areas of her scalp, and she began to lose some excess weight. PMID- 9746691 TI - Diagnosing and Managing Breast Disease During Pregnancy and Lactation. AB - Although carcinoma of the breast complicates 1:3000 deliveries in the US, most breast conditions unique to pregnancy and lactation are benign--for example, lactating adenoma, galactocele, gigantomastia, and benign bloody nipple discharge. Nevertheless, malignancy must be excluded by a thorough work-up, including breast biopsy if indicated; "watchful waiting" when a breast mass is discovered is no more appropriate than in a nonpregnant patient. During lactation, the major problems encountered often are part of a spectrum of inflammatory and infectious complications. Nasopharyngeal organisms from the infant are usually the source of breast infections in lactating women. Keeping the breast empty of milk promotes healing by helping to drain the culture medium that is facilitating growth of organisms. Hence, the earlier recommendations that breast-feeding cease during mastitis have been superseded by the knowledge that breast-feeding is generally not harmful to the infant and may speed resolution of the infectious process. The diagnosis and management of pregnancy-associated breast cancer (PABC) is reviewed. Pregnancy-associated masses are usually discovered by patient self-examination, and the clinician should proceed to fine needle aspiration or biopsy, rather than mammography, which has poor sensitivity during pregnancy and lactation because of increased breast density. Management of a new breast mass in pregnancy should maximize diagnostic accuracy and minimize the chances of missing PABC, yet avoid harm to the fetus or interruption of lactation. PMID- 9746692 TI - Genital Herpes: Treatment Guidelines. AB - Genital herpes, usually caused by herpes simplex virus type 2 (HSV-2), is the most common cause of genital ulceration. The primary episode of genital herpes is generally the most painful. Subsequent recurrences are generally milder and localized. Diagnosis is made clinically, but should be confirmed by culture or serology. Management includes antiviral drug therapy--acyclovir, valacyclovir, or famciclovir--as well as analgesics. In addition, patient counseling and education are vital. Antiviral treatment decreases the severity and duration of primary genital herpes and of recurrences, and it may be used as a continuous suppressive therapy to decrease the incidence of recurrence. Pregnant women who have a history of genital herpes or recent primary infection should deliver by cesarean section in the presence of genital lesions at labor or primary HSV infection at any time during the third trimester to prevent transmission to the neonate. Part 1, "Genital Herpes: Recognizing the Problem," addresses the problems involved in diagnosing the infection and quantifying the epidemic. PMID- 9746693 TI - Examining the Complex Relationship of Human Papillomavirus to Cervical Dysplasia and Carcinoma. AB - Human papillomavirus (HPV) infection is quite common in women and is clearly the major risk factor for cervical intraepithelial neoplasia and invasive cervical cancer. HPV DNA has been detected in 80% to 90% of CIN 3 lesions and invasive cervical cancers. While the most common presentation is warts, or condylomata, many infections are detected only by Pap smear cytologic evidence. We still do not have a clear understanding of HPV latency, reactivation, subclinical infection without apparent disease, and the triggers or cofactors required for malignant transformation. More than 70 different strains of HPV have been identified, and specific subtypes have been associated with a greater risk of progression to dysplasia and cervical cancer. A better knowledge of the oncogenic mechanisms of HPV and improved diagnostic testing is critical to guide future therapeutic and preventive investigations. The Pap smear is used for initial screening; cytologic results suspicious for premalignancy or malignancy are subsequently evaluated by colposcopy and biopsy of suspicious lesions. Cervical cancer has been designated an AIDS-defining illness; in HIV-infected patients, the prevalence of HPV is 5 times that of the general population. Because the disease presents at a later stage in HIV-infected patients and is less responsive to treatment, close attention to timely Pap smears and appropriate follow-up is important in this population. Presently, early detection and aggressive treatment and follow-up of premalignant lesions offer the best approach to the prevention of cervical cancer. PMID- 9746694 TI - Identifying Ultrasound Markers for Down Syndrome. AB - Although the incidence of Down syndrome increases with advancing maternal age, the use of maternal age alone as a screening tool results in the identification of only about one third of the cases of fetal Down syndrome. Screening tools for Down syndrome (trisomy 21) have become more sensitive and specific during the last few years. The use of biochemical markers for the screening of patients with fetuses having chromosomal anomalies has become more widespread in the obstetric community. The triple-screen test uses maternal age plus serum alpha-fetoprotein, unconjugated estriol, and human chorionic gonadotropin levels to calculate a risk for fetal Down syndrome. Because as many as 11% to 35% of fetuses with chromosomal defects have anatomical characteristics that can be visualized on a detailed ultrasound evaluation, researchers are studying the usefulness of this imaging technique as a screening tool for Down syndrome. Ultrasound findings associated with trisomy 21 may be divided into 2 groups. The first group comprises the common major malformations associated with Down syndrome, such as duodenal atresia and cardiac disease. The second group comprises the ultrasound screening indicators, that is, anatomical malformations highly specific to Down syndrome. This group includes brachycephaly, mild ventriculomegaly, macroglossia, abnormal facies, nuchal edema, echogenic or hyperechoic bowel, pyelectasis, and shortening of the limbs. Although the diagnosis of chromosomal anomalies remains dependent on karyotyping, the use of ultrasound may limit the number of invasive procedures and allow for more accurate genetic counseling of the mother at risk for delivering an infant with Down syndrome. PMID- 9746695 TI - Cervical Carcinoma in Pregnancy: Assessing the Diagnostic and Therapeutic Options. AB - Invasive cervical cancer is an infrequent complication of pregnancy, but it remains a significant cause of morbidity worldwide. When cervical cancer is suspected or diagnosed during pregnancy, the clinician must confront the potential risk to both the mother and the fetus. Due to the relative infrequency of this condition, guidelines for management are not clearly defined. Over the last several years, some investigators have reported on and advocated a more conservative approach to the management of this disease. This report assesses the recent literature on the evaluation and management options of invasive cervical cancer detected during pregnancy. Although delay of therapy may allow for maturation of the fetus, the data are currently insufficient to recommend a prolonged delay in treatment of the pregnant woman for advanced or even stage I disease. However, the available literature regarding a carefully planned, brief delay in therapy is encouraging, and all options should be presented to the patient. Ultimately, the management of invasive cervical cancer during pregnancy requires collaboration on the part of the patient, her family, and the entire multidisciplinary health care team, including the obstetrician, neonatologist, gynecologic oncologist, radiation oncologist, and social and psychiatric support personnel. PMID- 9746696 TI - Gynecology Case Challenge - Progressive Pelvic Pain in a Young Nulliparous Female. AB - A 28-year-old nulliparous white female is referred to you for evaluation of progressive lower abdominal pain associated with stretching exercise and with intercourse. A suspicious cervical lesion is discovered on pelvic exam. What is your diagnosis and treatment plan? PMID- 9746697 TI - Innovations in Breast Imaging: How Ultrasound Can Enhance the Early Detection of Breast Cancer. AB - Advancements in ultrasound technology have expanded the uses for ultrasonography in the evaluation of the breast. Breast masses can be delineated by specific ultrasonographic characteristics that allow them to be categorized according to their relative risk of being malignant. When combined with physical examination and mammography, breast ultrasonography can decrease additional radiation exposure associated with repeat mammograms. It also can lower the cost of evaluation of the breast, and it often reduces the number of open biopsies. In addition, ultrasound can be used for definitive pathologic diagnosis by guiding fine-needle aspiration and core biopsy, as well as facilitating preoperative needle localization for excisional biopsy. This article will review the expanding indications and describe the principles behind the performance and interpretation of breast ultrasonography. PMID- 9746698 TI - Perinatal Thyroid Dysfunction: Prenatal Diagnosis and Treatment. AB - Thyroid diseases affect up to 10% of women, but most respond well to treatment. During pregnancy, however, normal metabolic changes may obscure pathology, and improper management may harm the fetus. Tests for levels of thyroid stimulating hormone (TSH), free thyroxine, and free triiodothyronine are essential. Generally, high TSH values suggest primary hypothyroidism, while suppressed levels indicate hyperthyroidism. Hyperthyroidism is commonly manifested by goiter, ophthalmopathy, proximal muscle weakness, tachycardia, and weight loss or inability to gain weight. Among women, the most common etiology of thyroid disease is thyroid autoimmunity (Graves' disease or Hashimoto's thyroiditis); affected women are at an increased risk of postpartum thyroid dysfunction. Women who have Graves' disease diagnosed during pregnancy typically have a history of hyperthyroidism symptoms antedating conception, and occasionally thyroid stimulating immunoglobulins may be elevated enough to induce fetal hyperthyroidism. Women with Hashimoto's thyroiditis typically are euthyroid but may be hypothyroid with diffuse goiter; diagnosis is confirmed by elevated levels of antithyroid peroxidase antibodies or antimicrosomal antibodies. Other forms of thyroid dysfunction include benign or malignant nodules and hyperemesis gravidarum. Hypothyroidism typically is treated with levothyroxine. Hyperthyroidism is treated with antithyroid drugs. The goal is to avoid overdosage of medication, which could cause goiter and/or hypothyroidism in the fetus. PMID- 9746699 TI - Evaluating and Managing Postpartum Thyroid Dysfunction. AB - Postpartum thyroid dysfunction (PTD) occurs in approximately 5% to 10% of all women within 1 year following delivery and is usually due to intrinsic thyroid disease rather than hypothalamic or pituitary lesions. The most common etiology of PTD, which may resemble postpartum depression, is autoimmune thyroid disease (chronic or Hashimoto's thyroiditis). Women with Graves' disease who experience symptom exacerbation in the postpartum period account for a small percentage of cases. Clues to PTD include nonspecific symptoms such as tiredness, fatigue, depression, palpitations, and irritability. On physical examination, tachycardia may be noted. Goiters are detected in the majority of cases. The disease course varies; most patients experience a phase of hypothyroidism that takes 2 to 6 months to resolve, but some develop permanent hypothyroidism within 5 years of the diagnosis. PMID- 9746700 TI - Preterm Birth: The Role of Infection and Inflammation. AB - Preterm birth is the leading preventable cause of neonatal morbidity. Evidence shows that common genitourinary infections, which can easily be treated, cause large numbers of babies to be born prematurely. Because of their biologically immature organs, these newborns require intensive neonatal care, which leads to excess hospital costs early in life (approximately $3000/day at the University of Colorado). Long term, these children require follow-up for a range of disabling conditions, such as cerebral palsy, mental retardation, blindness, and/or deafness. Inexpensive screening during pregnancy can detect such common infections as bacterial vaginosis, trichomoniasis, chlamydia, and urinary tract infection; prompt treatment of these infections can effectively reduce admissions for preterm labor evaluation and can lower preterm birth rates. Bacterial vaginosis, in particular, has been consistently associated with a significantly increased risk of preterm births. Selective use of antibiotics in women during preterm labor and premature rupture of membranes significantly reduces both preterm birth rates and the risk of complications--in particular, from group B streptococcus (GBS) infection--in both babies and mothers. Implementation of appropriate screening and treatment of bacterial vaginosis and other prevalent infections can dramatically reduce the excess morbidity and mortality of infants "born too soon" because of reproductive tract infection. PMID- 9746701 TI - Gynecology Case Challenge - Spotting and Abnormal Cervix in a Young Woman on Oral Contraceptives. AB - A 22-year-old primiparous woman on oral contraceptives is referred to you for "spotting after intercourse" and an "abnormal-appearing cervix." What is your diagnosis and treatment plan? PMID- 9746702 TI - Percutaneous Bladder Neck Stabilization for Stress Urinary Incontinence in Women: The Technique, Risks, Benefits. AB - Recent reviews have noted failures of transvaginal surgical procedures designed to cure stress urinary incontinence in women. Modifications continue to be applied to improve the transvaginal approach, including anchoring the supporting sutures to the pelvic bones and reducing the transvaginal dissection to prevent further prolapse. This article reports the outcomes of 2 procedures: a bone anchoring technique with preservation of the endopelvic fascia in transvaginal suspension surgery for incontinence caused by urethral hypermobility, and a modified sling procedure for incontinence caused by intrinsic sphincteric deficiency. Results of the bone-anchor suspension showed an 81.7% cure rate in 71 patients who were followed for at least 3 years. Findings of the in situ sling procedure with bone anchoring showed a 97.5% cure rate in 40 patients who were followed for at least 2 years. The use of this bone-anchoring technique with preservation of the endopelvic fascia and the use of the modified sling technique appear to enhance the success rate without increasing the risk to the patient. As minimally invasive procedures, they also reduce surgical time and length of hospitalization, thus lowering costs. PMID- 9746703 TI - Weighing The Options In Medical Abortion. AB - For women who require interruption of pregnancy, medications that can offer a safe alternative to surgery--for example, methotrexate (though interruption of pregnancy is not an approved indication) and mifepristone (formerly known as RU486 and currently pending approval by the Food and Drug Administration [FDA])- are being examined. As with any new therapy, clinicians who wish to be able to offer this option should become familiar with the medications' safety and efficacy profiles and how they work, which patients are acceptable candidates, how the procedure is performed, how to counsel women seeking information, and how to manage complications. Nonsurgical abortion using either methotrexate or mifepristone in combination with misoprostol can be a safe and effective alternative to surgical abortion if a woman is no more than 7 weeks pregnant, clinical guidelines are followed, and access to surgical abortion is available for complications. Administration of either agent is followed within days by administration of misoprostol, a prostaglandin that induces uterine contraction and expulsion of the uterine contents. Side effects of all 3 agents are generally mild, but complications may include ongoing pregnancy, incomplete abortion, or excessive bleeding. PMID- 9746704 TI - Recognizing and Managing the Oral Clues That Point to Sjogren's Syndrome. AB - Sjogren's syndrome (SS), a chronic autoimmune exocrinopathy, occurs mainly after age 40. Most SS patients--80% to 90%--are women. SS is characterized by dry eyes and mouth due to lacrimal and salivary gland lymphocytic infiltration. It may be primary or secondary in association with a connective tissue disease, usually rheumatoid arthritis. Lymphoproliferation may produce extraglandular manifestations in pulmonary, cardiac, genitourinary, vascular, and/or nervous systems. The risk of lymphoma is increased 40-fold among SS patients. Dry mouth, or xerostomia, hinders eating, speaking, and swallowing. A thorough patient history, serum analysis, and salivary function tests are essential to determine the genesis of the xerostomia. Insufficient salivary protection can cause rampant dental destruction and soft-tissue mycosis in the mouth. A biopsy of the minor salivary gland from the lower lip is used to detect hallmark inflammatory changes that confirm the diagnosis of SS. Therapy is symptomatic. Regardless of the cause of xerostomia, therapy has 3 fundamental aspects: preventive dental care, dietary counseling (reduction of sugar intake to avoid caries), and moisture replacement (including artificial salivas, frequent sips of water, and room humidifiers). Women taking xerostomic medications may need to lower the dose or substitute them with less xerogenic drugs if possible. Salivation can be stimulated by chewing gum, mints, or paraffin. Cracked lips are treated with petroleum. Dental flossing, supplemental fluoride, and dental appointments every 3 to 4 months are essential to control caries. Pilocarpine, a parasympathomimetic drug that increases salivation, has been found to reduce the severity of xerostomia from radiotherapy; multicenter trials in SS patients are ongoing. PMID- 9746705 TI - Identifying and Treating Depression in Women With Cancer: A Primary Care Approach. AB - In general, depressive disorders in the US are more common in women than in men. In women with cancer, approximately 20% to 25% experience clinically significant depression and/or anxiety at some point during the course of medical treatment. This report profiles the differential diagnosis of depressive disorders as well as special medical variables, treatment options, and follow-up considerations for women with cancer. A range of psychotherapeutic and somatic treatments are available, with selective serotonin reuptake inhibitor antidepressants being the mainstay of drug therapy. Despite an array of available treatments, depression in patients with cancer remains underdiagnosed and undertreated; it is imperative that these patients be treated for both of their diseases. Depression left untreated in women with cancer may not only cause significant emotional suffering but also slower medical recovery, less adaptive health behaviors, and a negative effect on medical outcome and, ultimately, on survival. Patients who do not respond to conventional treatment approaches should be referred to a consulting psychiatrist for confirmation of diagnosis and consideration of other treatment options. PMID- 9746706 TI - Diagnosing and Treating Breast Cancer in the Pregnant Woman. AB - Because breast cancer is an uncommon occurrence during pregnancy, symptoms and signs of the disease may be overlooked, resulting in delays in treatment and potentially compromising survival. In many cases, the physician may mistake signs of disease for the normal physiologic changes of pregnancy. For this reason, it is imperative that physicians perform careful clinical breast examinations in all pregnant patients--particularly early in gestation, before the breasts become difficult to examine. Upon finding any suspicious breast mass, an open biopsy without delay is indicated. Once a diagnosis of breast cancer is confirmed, a modified radical mastectomy is the treatment of choice in a pregnant woman because of the hazards of adjuvant therapy to the fetus. The administration of adjuvant therapy, such as radiation therapy or chemotherapy, may pose a risk to the fetus and requires careful consideration by the physician and patient. In some cases, especially when disease presents early in gestation, an interruption of the pregnancy may be warranted. Importantly, stage for stage, breast cancer during pregnancy has a similar prognosis to that of breast cancer in young, nonpregnant women; pregnancy itself does not appear to have an adverse effect on the disease process. In addition, with careful counseling, pregnancy subsequent to breast cancer is possible for some women with good prognoses. PMID- 9746707 TI - Hearing Loss: Does Gender Play a Role? AB - An estimated 8 million women in the US have difficulty hearing, and 2 million of those are able to hear, at best, only shouted words. Women of all ages have better hearing than men at frequencies above 2000Hz. The better pure-tone audiometry thresholds of women at high frequencies is paradoxically accompanied by a "gender reversal" in which women, as they age, have a poorer capacity to hear at low frequencies--specifically those below the 1000- to 2000-Hz range- than do men. Thus, the pattern of hearing loss with aging may differ between women and men. The hypothesized role of ovarian hormones and cardiovascular disease in hearing loss is reviewed, and such interventions as cochlear implants to correct hearing loss in women are highlighted. Research into organ of Corti hair-cell regeneration is ongoing and may offer recovery of hearing in the next century. PMID- 9746708 TI - From GnRH to SSRIs and Beyond: Weighing the Options for Drug Therapy in Premenstrual Syndrome. AB - Pharmacologic intervention is now the most effective therapy available for treating premenstrual syndrome (PMS). Although there are still no Food and Drug Administration-approved medications for this indication, several well-designed studies have been conducted, the results of which may guide the clinician's treatment of women with this disorder. Consequently, less-proven nonpharmacologic modalities, such as dietary modification, exercise regimens, and psychotherapy, are more readily supplanted by the use of medication. Three classes of agents have been shown to have varying degrees of effectiveness in relieving PMS symptoms and are increasingly being used to treat the disorder: gonadotropin releasing hormone (GnRH) agonists, benzodiazepines, and selective serotonin reuptake inhibitors (SSRIs). While the GnRH agonists like leuprolide acetate, nafarelin acetate, or goserelin acetate have been shown to be highly effective in select cases, their side effects relegate them to use in patients who are unresponsive to other agents. More recently, the benefits of alprazolam (a benzodiazepine) and the SSRIs (especially fluoxetine) have been definitively established. In addition to these medications used to treat premenstrual syndrome in general, other drugs that are used to treat specific aspects of the disorder include danazol for headaches and spironolactone for fluid retention. PMID- 9746709 TI - Tubal Occlusion Failures: Implications of the CREST Study on Reducing the Risk. AB - Through data reported in the US Collaborative Review of Sterilization (CREST) study, we have learned that 10-year cumulative failure rates of sterilization done by tubal occlusion are much higher than originally thought. While the small, earlier studies reported failure rates as low as 3 to 4 per 1000 procedures, they often followed women for only 2 years after the procedure. When pregnancies occurred during this period, the operative assumption was that these failures were due to incomplete occlusion. Most reports have not addressed the possibility of recanalization leading to failures. The CREST findings, however, suggest that failure rates are closer to 18 per 1000, depending on the occlusion method used and characteristics of the patient. This study also shed light on the factors that increase the risk of ectopic pregnancy after sterilization procedures. These new long-term data indicate that all providers should know that pregnancy, including ectopic pregnancy, can occur in women with history of tubal occlusion for sterilization, especially many years after the original procedure. PMID- 9746710 TI - Cervical Cancer Prevention: Toward Cost-Effective Screening. AB - The decrease in the incidence of invasive cervical cancer has been credited to the widespread use of Papanicolaou (Pap) smear screening. Although relatively inexpensive to perform, Pap smears, if positive, often result in further diagnostic work-up (eg, colposcopy, biopsy, endocervical curettage) and associated patient anxiety. Unfortunately, false positives are frequent with Pap smears, and even screened populations of patients continue to have a significant incidence of cervical cancer. Presumably, expanding screening programs to unscreened populations or screening selected, at-risk populations more frequently could further reduce the incidence of invasive cervical cancer. Yet, few rigorous, prospective studies exist to allow for the formulation of cost effective guidelines that optimize screening resources. To determine just how much screening is cost-effective, the medical community will have to answer several questions regarding the definition of cost-effectiveness itself, the optimal age to begin screening, whether abnormal Pap smears can be better stratified according to risk, the limitations of Pap smear screening, and whether advances in technology can help increase the positive predictive value of current screening strategies. PMID- 9746711 TI - Periareolar Breast Abscess: Redefining the Disease and Its Treatment. AB - Periareolar breast abscess has been an elusive condition, with much debate about its etiology over the last several decades. Presenting symptoms include nipple discharge, mastalgia, and recurrent abscesses with draining fistulas. Many experts disagree about whether this condition develops when inflammation of the duct leads to dilation or whether it begins with dilation that leads to inflammation. Because the frequency of asymptomatic dilated ducts found incidentally in patients during surgery or upon autopsy exceeds that of patients with symptomatic duct dilation or ectasia, we believe that mechanical obstruction with associated retention of secretions is at the core of this disease process. In this article, we term and characterize mammary-duct-associated inflammatory disease as a 3-phase pathologic process that leads to recurrent nonlactational periareolar breast abscess in nonpuerperal women. Effective treatment of abscesses should be based on the disease's pathogenic process and should include excision of all involved ducts. Treated by this method, patients appear to experience minimal sequelae and low recurrence of abscesses. PMID- 9746712 TI - Clinician's Photo Guide To Recognizing and Treating Skin Diseases in Women: Part 1. Dermatoses Not Linked to Pregnancy. AB - The clinical presentation of certain dermatologic conditions differs between women and men; this may be especially true when women are perimenstrual or pregnant. Skin diseases that erupt or become aggravated during the perimenstrual period include autoimmune progesterone dermatitis and melasma. Dermatologic conditions that may be exacerbated perimenstrually include acne vulgaris, rosacea, lupus erythematosus, psoriasis, atopic eczema, lichen planus, dermatitis herpetiformis, erythema multiforme, and urticaria. The hormonal effects of increased cutaneous vascularity, seborrhea, and dermal edema during the perimenstrual period may account for the eruption of or increase in severity of these diseases. Clinical presentation, differential diagnoses, and treatment options for select cutaneous conditions are discussed. PMID- 9746714 TI - Clinician's Photo Guide To Recognizing and Treating Skin Diseases in Women: Part 2. Pregnancy-Related Dermatoses. AB - This report identifies 4 pregnancy-induced dermatoses: (1) pemphigoid gestationis, (2) polymorphic eruption of pregnancy, (3) prurigo of pregnancy, and (4) pruritic folliculitis of pregnancy. According to 1 study of 3192 pregnancies, 0.06% of the women had pemphigoid gestationis (PG), 0.5% had polymorphic eruption of pregnancy (PEP), 0.2% had prurigo of pregnancy (PP), and 0.03% had pruritic folliculitis of pregnancy (PFP). Some reports have suggested an increased risk of fetal morbidity and mortality, as well as an increased risk of premature births, among women with PG. The incidence of fetal morbidity and mortality for the other dermatoses of pregnancy appears to be similar to that in normal pregnancies. Among the drugs used in dermatology, isotretinoin and antineoplastic agents, such as methotrexate, are 2 types that present high risk during pregnancy. Antipruritic medications, such as trimeprazine and doxepin, and some nonsteroidal anti-inflammatory agents, such as indomethacin, also should be avoided during pregnancy and lactation. Analgesics, including acetaminophen, are associated with minimal risk to the fetus or infant. Use of topical corticosteroids is associated with a low risk during pregnancy. Fortunately, many dermatologic disorders allow deferral of treatment or alternate therapeutic methods during pregnancy. PMID- 9746715 TI - A 35-Year-Old Asymptomatic Woman With ASCUS on Pap Report. PMID- 9746716 TI - Haemophilus influenzae Genome: After Sequencing, Then What? AB - Haemophilus influenzae is an important human pathogen that causes a number of chronic and acute infections--notably, life-threatening bacteremia and meningitis in infants and community-acquired pneumonia in adults. The technological feat of sequencing the entire H influenzae genome was completed less than a year ago. Now that the genetic instruction manual is available, the task of deciphering its meaning has begun. This search will be a lengthy one, but it holds the promise of finding new ways to prevent, detect, and treat disease. The information gained will also shed light on mechanisms used by other bacterial pathogens and help to increase our understanding of how they cause disease. PMID- 9746717 TI - Genital Herpes: Recognizing the Problem. AB - Genital herpes, usually the result of an infection with herpes simplex virus type 2 (HSV-2), is the most common cause of genital ulceration. The first clinical episode is called primary genital herpes; recurrent HSV infection occurs in up to 80% of patients. Because not every individual who acquires the virus develops symptoms, it is difficult to estimate the prevalence of HSV infection. Transmission of virus occurs not only in the presence of apparent lesions, but asymptomatic viral shedding can also spread infection in the absence of lesions. The primary episode of genital herpes is generally the most painful, characterized by multiple and bilateral lesions and associated with tender inguinal lymphadenopathy as well as systemic symptoms. Subsequent recurrences are generally milder and localized. Diagnosis is made clinically, but should be confirmed by culture or serology. Part 2, "Genital Herpes: Treatment Guidelines," addresses aspects of treatment, including special considerations in treating pregnant women. PMID- 9746718 TI - Human endogenous retroviral sequences: possible roles in reproductive physiopathology. PMID- 9746719 TI - Developmental expression of the androgen receptor during virilization of the urogenital system of a marsupial. AB - In the marsupial tammar wallaby, virilization begins approximately 3 wk after the onset of testosterone synthesis. In the eutherian mammal, in contrast, the onset of virilization of the male urogenital tract occurs shortly after the onset of androgen synthesis. Androgen action requires the presence of the androgen receptor to mediate a response in target tissues. We therefore investigated the developmental expression of the androgen receptor (AR) in both sexes of the tammar wallaby. AR gene transcript was detected in fetal gonad and brain as early as Day 19 of the 26.5-day gestation, 7 days earlier than the first rise in testicular testosterone (Days 0-5 postpartum [p.p.]). Immunoreactive AR was identified in the male urogenital sinus (UGS) 2 days before birth and in the female UGS and mammary glands by the day of birth. AR was present in the UGS, vagina, and prostate until Day 152 p.p., the oldest age examined. AR was identified in the gubernaculum testis at Day 2 p.p. and became more abundant by Day 32. In the phallus of both sexes, AR was identified by Day 4 p.p. and until Day 157, the oldest age examined. AR was not detected in the scrotum at any age from the day of birth to Day 157. Maturation of the phallus, wolffian duct, and epididymis was marked by appearance of epithelial immunostaining. AR was localized in the epithelium of the UGS in females by Day 50 p.p. but was not found in the epithelium of the male UGS up to Day 152 p.p., the oldest examined. AR were found in the mesenchyme of the UGS of male and female tammars 3-4 wk before virilization is first evident in the male at Day 25 p.p. We conclude that the presence of AR is not the initiating signal for virilization of the UGS in this marsupial male. PMID- 9746721 TI - Large-format, two-dimensional polyacrylamide gel electrophoresis of ovine periimplantation uterine luminal fluid proteins: identification of aldose reductase, cytoplasmic actin, and transferrin as conceptus-synthesized proteins. AB - Early pregnancy in ruminants, such as the sheep, is characterized by relatively extensive development of the conceptus before attachment to the endometrium. Between the period of blastocyst hatching and initial attachment, the uterus responds to signals from the conceptus and adapts to provide an environment that permits the establishment of pregnancy. We used large-format two-dimensional (2D) PAGE to analyze the dynamic changes in protein composition of uterine luminal fluid (ULF) during this stage of pregnancy, and we determined the contribution of each of the extraembryonic membranes and the endometrium to these changes. The majority of the more than 40 pregnancy-associated proteins in ULF at Day 17 were secreted by the conceptus. By 2D gel map comparison and Western blotting, we identified transferrin, secreted by the yolk sac from Day 15, and cytoplasmic actin, one of the most abundant proteins produced by the trophoblast at Day 17. Apolipoprotein A1 and aldose reductase, whose abundance were markedly increased in pregnancy, were identified by peptide microsequencing. Aldose reductase, an enzyme required for the conversion of glucose to fructose, was shown to be synthesized by the trophoblast, and its detection even before the formation of the placenta suggests that the synthesis of fructose may occur much earlier than previously reported. PMID- 9746720 TI - Progesterone-induced acrosome reaction in stallion spermatozoa is mediated by a plasma membrane progesterone receptor. AB - The aim of the present study was to investigate whether the induction of stallion sperm acrosome reaction (AR) by progesterone is mediated by binding of progesterone to a receptor on the sperm plasma membrane or to an intracellular progesterone receptor. Progesterone-BSA conjugate labeled with fluorescein isothiocyanate (P-BSA-FITC) in combination with a vital stain, ethidium homodimer, was applied to visualize the presence of the progesterone receptor on living spermatozoa. Alternatively, an indirect immunofluorescence technique employing a monoclonal antibody (C-262) against human intracellular progesterone receptor was conducted to validate the presence of the progesterone receptor. Immunogold labeling techniques enabled ultrastructural localization of P-BSA-FITC or C-262 with transmission electron microscopy. The dynamic changes in labeling patterns were monitored for sperm cells, using fluorescence microscopy and flow cytometry during a 5-h capacitation period. An increasing number of viable cells showed affinity for P-BSA-FITC or C-262 at the acrosomal plasma membrane region of the sperm head, while a decreasing number of viable cells were not labeled. In contrast, almost all deteriorated cells were labeled in the cytosol of the postequatorial region of the sperm head. Incubation with P-BSA-FITC resulted in the induction of AR but to a lesser extent than that for sperm incubated with free progesterone. Therefore, coupling of progesterone to its receptor on the sperm plasma membrane appears to be an important step in the induction of the AR. PMID- 9746722 TI - Antioxidant systems in rat epididymal spermatozoa. AB - Mammalian caput and cauda epididymidal spermatozoa exhibit diverse stages of maturation, and their plasma membrane shows diverse composition and stability levels, thus enabling these spermatozoa to undergo the acrosomal reaction after transit through the epididymis. As a result, the study of antiperoxidative mechanisms is quite relevant, since epididymal spermatozoa must be properly protected against agents such as reactive oxygen species, which can impair the complex maturation process. We considered activities of certain enzymes (glutathione peroxidase [GPx], phospholipid hydroperoxide glutathione peroxidase [PHGPx], glutathione reductase [GR], superoxide dismutase [SOD], and catalase [CAT]) and the vitamin E content in isolated rat caput and cauda epididymidal spermatozoa. The results indicate that caput epididymidal sperm have significantly greater PHGPx (3.5x), GPx (2.4x), and SOD (1.7x) activities, as well as a greater amount of vitamin E (3.8x). There were no detectable differences in the GR and CAT activities of caput and cauda epididymidal spermatozoa. The substantial drop in PHGPx activity during epididymal transit is discussed in relation to an additional function of this enzyme: the use of caput sperm protamines as a sulfhydryl substrate. In vitro peroxidation of the two sperm populations by the free radical generator (azo-initiator) 2,2'-azobis(2 amidinopropane) dihydrochloride revealed that only about 13% of the vitamin E content of the caput epididymidal spermatozoa was consumed, which contrasts with the greater consumption (about 70%) of the vitamin in cauda epididymidal spermatozoa. Selective inhibition of PHGPx, SOD, or CAT did not change this picture. The higher susceptibility of cauda epididymidal spermatozoa to radicals is discussed in relation to the diverse enzymatic activities, vitamin E content, and peroxidative response. These factors are correlated with the different stages of sperm cell maturation, which are characterized-from caput to cauda epididymidis-by progressive destabilization of the plasma and acrosomal membranes. PMID- 9746723 TI - Human endothelial cell migration is stimulated by urokinase plasminogen activator:plasminogen activator inhibitor 1 complex released from endometrial stromal cells stimulated with transforming growth factor beta1; possible mechanism for paracrine stimulation of endometrial angiogenesis. AB - Human endometrial stromal cell cultures, stimulated for two days with recombinant transforming growth factor beta1 (TGFbeta1; 10 ng/ml), contained conditioned medium concentrations of urokinase plasminogen activator (uPA), plasminogen activator inhibitor 1 (PAI1), and uPA:PAI1 complex. Since a number of cellular effects have been reported to follow a binding of enzymatically inactive uPA to the receptor in different cell types, we studied the influence of uPA:PAI1 complex on human umbilical vein endothelial cells (HUVEC) and human microvascular endothelial cells (HMEC-1). Increasing concentrations of uPA:PAI1 complex as well as free uPA resulted in a dose-dependent stimulation of endothelial cell migration. Stimulation by the complex was of the same magnitude as that of free uPA on a molar basis and reached its maximum at 1 nM in both cell types. PAI1 by itself, however, had no effect on cell migration. The migratory response to both uPA and the uPA:PAI1 complex was inhibited by antibody adhesion to the cell surface receptor for uPA. In addition, we found that TGFss1 had a direct stimulatory effect on migration in both HUVEC and HMEC-1. This response did not, however, involve the binding of uPA to the uPA receptor. Since TGFbetas are expressed in endometrial tissue and reportedly stimulate angiogenesis in other tissues in vivo, though not endothelial cell proliferation in vitro, they may engage in the regeneration of endometrial vasculature indirectly via perivascular cells. We found that the uPA:PAI1 complex, when released from endometrial stromal cells in response to TGFbeta1, stimulated endothelial cell migration. This suggests a possible mechanism for paracrine stimulation of endometrial angiogenesis. PMID- 9746724 TI - Major proteins of bovine seminal plasma and high-density lipoprotein induce cholesterol efflux from epididymal sperm. AB - One of the hypotheses to explain the mechanism of capacitation involves the loss of sperm membrane cholesterol. Here, we studied whether or not the major proteins of bovine seminal plasma designated as BSP-A1, -A2, -A3, and -30-kDa (collectively called BSP proteins), which are implicated in sperm capacitation, induce cholesterol efflux. When epididymal sperm were labeled with [3H]cholesterol and incubated with bovine seminal plasma (0.05-2%) or BSP proteins (20-120 microg/ml) for 8 h, the sperm lost [3H]cholesterol (3.6-fold and 3-fold, respectively). The same results in the presence of BSP-A1/-A2 were obtained (3.5-fold) by direct determination of cholesterol on unlabeled epididymal sperm. Analysis of efflux particles by ultracentrifugation on a sucrose gradient revealed a single symmetrical peak of radioactivity at 1.14 g/ml. Immunoblotting of the fractions obtained from size-exclusion chromatography of the efflux particles showed that a portion of the BSP proteins were associated with [3H]cholesterol. Heparin (12 microg/ml) alone did not stimulate cholesterol efflux. In contrast, high-density lipoprotein (HDL, 100 microg/ml) alone stimulated cholesterol efflux up to 3.1-fold after 8 h. When labeled epididymal sperm were preincubated for 20 min with BSP-A1/-A2 (120 microg/ml), washed, and incubated with HDL (100 microg/ml) for 8 h, the total cholesterol efflux of the sperm suspension was 51.8 +/- 5.0% compared to 39.3 +/- 1.2% when HDL alone was used. These results indicate that BSP proteins and HDL play an important role in the sperm sterol efflux that occurs during capacitation. Furthermore, the heparin induced sperm capacitation did not involve the efflux of sperm membrane cholesterol. PMID- 9746725 TI - Effect of calf isolation on follicular wave dynamics, gonadotropin and metabolic hormone changes, and interval to first ovulation in beef cows fed either of two energy levels postpartum. AB - The effects of postpartum energy intake, restricted suckling, and cow-calf isolation on concentrations of LH, FSH, growth hormone, and insulin-like growth factor-I (IGF-I) and on postpartum anestrous interval were determined by randomly allocating beef cows with a mean body condition score of 2.3 +/- 0.1 to receive either 80 MJ metabolizable energy (low-energy diet [L]; n = 51) or 120 MJ metabolizable energy (high-energy diet [H]; n = 52) per cow per day from calving. At 30 days postpartum, cows within diet were randomized to 1) have continued full access to their calves from birth to weaning (ad libitum suckling: ADLIB), 2) be suckled once-daily with their calves penned adjacent (restricted suckling, adjacent: RESADJ), 3) be isolated from all calves except for a once-daily suckling period (restricted suckling, isolated: RESISO). The mean postpartum interval was similar (p > 0.10) for L and H cows (62 and 63 days, respectively). RESADJ cows had a shorter (p < 0.05) postpartum interval than ADLIB cows, and RESISO cows had a shorter interval (p < 0.05) than RESADJ cows, with all effects independent (p > 0.10) of diet. FSH secretion pattern was not affected by diet, suckling treatment, sequential follicle wave number, or follicle wave retrospectively realigned to emergence of first ovulatory wave. Within 5 days of suckling restriction and calf isolation, the number of LH pulses increased from 0.18 to 0.48 pulses per hour (p < 0.05). Both mean LH and the mean number of LH pulses increased linearly (p < 0.01) during the six follicle waves up to the first ovulatory wave. From 80 days before, until the time of, first ovulation, growth hormone decreased (p < 0.05) while IGF-I increased (p < 0.05), irrespective of treatment. The results indicate that the "suckling effect" in beef cows is the major factor affecting the duration of the postpartum interval and suggests that the maternal bond is more important than suckling in regulating LH pulse frequency, the key endocrine factor determining whether or not a dominant follicles ovulates. Removal of the suckling effect resulted in a rapid increase in LH pulse frequency, which was not dependent on level of postpartum nutrition, at least within the nutritional limits of this study. Mean concentrations of FSH, unlike LH, did not vary with follicle wave number, suggesting that lack of FSH is not a major factor delaying the resumption of ovulation in postpartum beef cows. PMID- 9746726 TI - Effects of the estrous cycle and early pregnancy on uterine expression of Mx protein in sheep (Ovis aries). AB - Conceptuses of ruminant ungulates produce large amounts of a type I interferon, interferon-tau (IFNtau), which is the signal for maternal recognition of pregnancy. Induction of cellular Mx proteins is an important component of the response to type I interferon in the immune system, but Mx regulation and function have not been studied in the uterus. This study examined temporal and spatial alterations in ovine uterine Mx expression during the cycle and early pregnancy using immunohistochemistry, in situ hybridization, and Northern and slot-blot analysis. Sheep uterine endometrium expressed a single approximately 2.5-kilobase Mx mRNA transcript that was detectable at all stages of the estrous cycle and early pregnancy examined. In cyclic ewes, mRNA abundance in endometrium increased from Day 1 to peak levels at Day 13 and then declined to Day 15. In pregnant ewes, steady-state levels of Mx mRNA were first detected above the level in cyclic ewes at Day 13 postmating, were greater than 10-fold higher at Day 15, and remained elevated at Day 19. Expression of Mx mRNA in the myometrium did not change during the estrous cycle but increased approximately 23-fold between Days 11 and 15 of pregnancy. Immunohistochemical and in situ hybridization analysis revealed a similar temporal pattern of Mx expression. In cyclic ewes, Mx protein and mRNA were initially localized to the luminal epithelium at Days 1 and 3, increased from Days 5 to 13, especially in the shallow uterine glands, and then declined at Day 15. Pregnancy resulted in up-regulation of Mx expression in the luminal and glandular epithelium, stroma, and myometrium. Punctate Mx immunostaining and Mx mRNA concentrations were greatest when progesterone production was maximal during the estrous cycle and were strongly up-regulated by the conceptus across the entire uterine wall. It is suggested that a cascade of induction of Mx gene expression proceeds from the luminal epithelium to the outer longitudinal myometrium and that transcriptional activation of the promoter may involve both soluble cytokines (i.e., IFNtau) and steroid hormones (i.e., progesterone). PMID- 9746727 TI - Role of relaxin and estrogen in the control of eosinophilic invasion and collagen remodeling in rat cervical tissue at term. AB - Ripening and dilation of the rat cervix at term involves a widespread reduction in the density and organization of collagen fibers following a polymorphonuclear eosinophilic invasion. These are hormonally regulated events; however, the correlation between hormonal milieu and these morphological changes is not well understood. To investigate the role of preparturient relaxin and estradiol-17ss (E2) in cervical collagen remodeling and eosinophilic infiltration, pregnant rats were either sham-ovariectomized (group C) or ovariectomized in the morning of Day 22. Ovariectomized rats were treated with E2 (group OE), relaxin (group OR), E2 plus relaxin (group OER), or hormone vehicles (group O). There were 4 or 5 animals per group. Cervices were taken from animals killed approximately 1 h before expected parturition. Five-micrometer serial sections of paraffin-embedded cervices were stained with either Sirius Red in alkaline solution to measure eosinophil infiltration or in Picrosirius to measure collagen birefringence. Ovariectomized rats treated with E2 (group OE or OER) showed high levels of eosinophilic infiltration that did not differ from those in group C intact controls. However, the values of eosinophilic infiltration in ovariectomized rats treated with relaxin or hormone vehicles (groups OR and O) were low and far below (p < 0.01) those of groups OE and C. In ovariectomized rats treated with E2 alone (group OE), the widespread reduction in collagen organization that occurred in group C controls was not observed. It was only in ovariectomized rats treated with relaxin or E2 + relaxin (groups OR and OER) that the values of birefringence were low, and they were as low as in control group C. In conclusion, this study indicates that eosinophilic infiltration and collagen remodeling in the rat cervix at term are under the control of different hormones: E2 stimulates eosinophilic invasion, and relaxin promotes a widespread reorganization of collagen fibers. PMID- 9746728 TI - Growth factors and components for extracellular proteolysis are differentially expressed during in vitro maturation of bovine cumulus-oocyte complexes. AB - In the present study, the time-dependent collagenolytic potential and mRNA transcription of extracellular matrix (ECM)-degrading components, transforming growth factor beta1 (TGFbeta1), and both basic fibroblast growth factor (bFGF) and FGF receptors (FGFR) in bovine cumulus-oocyte complexes (COCs) were investigated during 24 h of in vitro maturation (IVM). COCs were collected from 2 to 6-mm follicles, cultured in maturation medium, and sequentially removed at 3 h intervals for analysis. From 285 oocytes matured under these conditions, 114 (40.0%) developed to blastocysts on Day 9 after fertilization. Gelatin zymograms revealed protease activity at about 55 kDa for COCs matured for at least 3 h and two additional proteolytic zones at about 70 kDa after at least 9 h of IVM. The mRNAs of ECM-degrading components urokinase-type plasminogen activator (uPA), plasminogen activator inhibitor 1 (PAI1), matrix metalloproteinase 1 (MMP1), and tissue inhibitor of metalloproteinases 1 (TIMP1), as well as TGFbeta1, bFGF, and FGFR, were detected during IVM in a factor-specific pattern: all transcript levels found at COC 0 generally increased after 3 h of maturation and either remained high or decreased thereafter. On the basis of the chosen reverse transcription-polymerase chain reaction technique, one could suggest relative higher mRNA concentrations for TIMP1, PAI1, and both growth factors compared to uPA, MMP1, and FGFR. Our results suggest a finely tuned extracellular proteolysis during IVM of bovine COCs, for which the concerted action of modulating growth factors like bFGF and TGFbeta1 may be essential. PMID- 9746729 TI - Characterization of inhibin/activin subunit, activin receptor, and follistatin messenger ribonucleic acid in human and mouse oocytes: evidence for activin's paracrine signaling from granulosa cells to oocytes. AB - Inhibin, activin, and follistatin (FS) are gonadal proteins that appear to have a role in regulating folliculogenesis through possible paracrine and/or autocrine interactions. To further examine the potential role of activin in oocyte granulosa cell communication, we developed a sensitive reverse transcription polymerase chain reaction protocol to analyze mRNA for the alpha, betaA, and betaB inhibin/activin subunits, FS, and the four activin receptor subtypes in individual human and mouse oocytes. The resulting expression pattern was further compared to that in human cumulus granulosa cells. Our results indicate that neither ssA nor betaB mRNA was detectable in any human or mouse oocyte, that alpha subunit was marginally present in some of the human oocytes, and that FS mRNA was detectable in human but not mouse oocytes. On the other hand, inhibin/activin subunit and FS mRNAs were abundantly expressed in cumulus cells. In addition, mRNAs for all four activin receptor subtypes (ActRIA, ActRIB, ActRIIA, and ActRIIB) were easily detectable in both oocytes and granulosa cells and appeared to be differentially expressed in oocytes during nuclear maturation. Finally, RNAs for both zona pellucida 3 and growth-differentiation factor-9, which were originally used as oocyte-specific markers, were detected in human but not mouse cumulus cells, although at lower levels than observed in oocytes. Taken together with previous studies, these results indicate that oocytes may be capable of responding to, but not synthesizing, activin. PMID- 9746730 TI - Effects of gonadotropin-releasing hormone pulse frequency modulation on the reproductive axis of photoinhibited male Siberian hamsters. AB - In Siberian hamsters, photostimulation evokes differential release of the gonadotropins, with FSH rising rapidly and LH levels rising much later. We have tested the hypothesis that differential release of gonadotropins in this species can be mediated by changes in the frequency of pulsatile GnRH stimulation. Photoinhibited Siberian hamsters received GnRH pulses at frequencies of 1 pulse every 45 (fast), 90 (medium), or 180 min (slow). Animals were killed at 0, 3, 5, 10, 20, and 30 days after treatment. There was a clear GnRH pulse frequency effect on LH release, with fast pulses > medium pulses > slow pulses > short-day (SD) controls. In addition, 10 days of fast-frequency GnRH pulses produced LH levels significantly greater than LH levels in animals exposed to 10 days of medium or slow GnRH pulse frequencies. Pulsatile GnRH produced the following serum FSH relationships: medium pulses > fast pulses > SD. The FSH response to slow GnRH frequency fell between the two faster frequencies. The effect of GnRH pulse frequency on paired testes weight was as follows: fast pulses = medium pulses > slow pulses > SD controls. The differing GnRH pulse frequencies produced the following testosterone relationships; fast pulses > medium pulses = slow pulses = SD controls. These results agree with studies showing that slower GnRH pulse frequencies facilitate FSH release, while faster GnRH pulse frequencies favor LH release. Our observations are also consistent with the idea that the singular release of FSH after transfer of hamsters to a long-day photoperiod is mediated by alterations in the frequency of endogenous pulsatile GnRH release. PMID- 9746731 TI - Estradiol-mediated suppression of apoptosis in the rabbit corpus luteum is associated with a shift in expression of bcl-2 family members favoring cellular survival. AB - In the rabbit, estradiol is the primary luteotropic hormone. Estradiol withdrawal results in a rapid decline in serum progesterone and eventually in corpus luteum (CL) regression. The objective of this study was to determine whether estradiol modulates luteal cell apoptosis. In the first experiment, rabbits were randomly assigned to one of five experimental groups. An empty capsule (control) or estradiol-filled Silastic capsule was inserted s.c. on Day 0 of pseudopregnancy (day of hCG administration). On Day 11 of pseudopregnancy, some of the group I (control) and group II (estradiol capsule) rabbits were subjected to laparotomy, and one ovary from each rabbit was perfused in vitro to determine progesterone secretion rates. The CL from the contralateral ovary were dissected, snap-frozen, and stored at -70 degrees C until analyzed for internucleosomal DNA cleavage (apoptosis). Estradiol-containing capsules were removed from some of the remaining rabbits on Days 8, 9, and 10 to initiate estradiol deprivation. Rabbits were then subjected to laparotomy 24, 48, or 72 h after capsule removal (groups III, IV, and V, respectively), and ovaries or CL were processed as described above. Deprivation of estradiol for 24 (group III), 48 (group IV), or 72 (group V) h in vivo reduced in vitro progesterone secretion rates by more than 90% as compared to that in ovaries collected from estradiol capsule-intact animals. After in vivo endogenous estradiol suppression, withdrawal of exogenous estradiol resulted in luteal cell apoptosis, which increased in a time-dependent manner. Northern blot analysis revealed an increase in bax mRNA levels and a decrease in bcl-x mRNA levels coincident with luteal cell apoptosis induced by estradiol withdrawal. These data demonstrate that changes in progesterone production caused by estradiol exposure and deprivation are in part related to luteal cell apoptosis, and alterations in the expression of bcl-2 gene family members may be one of the mechanisms by which estradiol exerts its luteotropic effect in the rabbit CL. PMID- 9746732 TI - Characterization of 16- to 20-kilodalton (kDa) connective tissue growth factors (CTGFs) and demonstration of proteolytic activity for 38-kDa CTGF in pig uterine luminal flushings. AB - Connective tissue growth factor (CTGF) is a growth and chemotactic factor for fibroblasts encoded by an immediate early gene that is transcriptionally activated by transforming growth factor ss. Although the primary translational product of the pig CTGF gene is predicted to be of approximate Mr 38 000, pig uterine luminal flushings (ULF) contained 10- to 20-kDa CTGF proteins that were heparin-binding and mitogenic, whereas 38-kDa CTGF was not apparent. The N termini of two microheterogeneous forms of 16-kDa CTGF, as well as 18-kDa and 20 kDa forms of CTGF, commenced at, respectively, Cys199, Ala197, Asp186, and Asp186 and did not correspond to intron-exon boundaries in the CTGF gene. Northern blotting revealed a single porcine (p) CTGF transcript of 2.4 kilobases in endometrium from Day 10 to 16 cycling or pregnant pigs. Ten- to twenty-kilodalton pCTGF proteins in ULF were stable for 48 h at 37 degreesC whereas native 38-kDa pCTGF was degraded within 10 min under the same conditions. CTGF-degrading activity in pig ULF was heat-sensitive and concentration- and time-dependent. Ten to twenty-kilodalton CTGF levels in ULF peaked on Day 16 of the cycle and on Day 12 of pregnancy and were highly correlated with the levels of proteolytic activity for 38-kDa CTGF. Collectively these data suggest that bioactive 10- to 20-kDa CTGF proteins are generated in utero through limited proteolysis of the 38 kDa CTGF primary translational product. PMID- 9746733 TI - Developmental changes in the expression of the growth hormone receptor messenger ribonucleic acid and protein in the bovine ovary. AB - By reverse transcription-polymerase chain reaction, the transcript of the growth hormone receptor (GHR) was demonstrated in oocytes, follicular cells, and corpus luteum of the bovine ovary. Immunoblotting using the monoclonal antibody mAb 263 resulted in a distinct protein band at 120 kDa, confirming that translation of the mRNA takes place in the same cells. Nonradioactive in situ hybridization revealed that distribution of the mRNA encoding GHR was correlated with the developmental stage of the follicle. Whereas in primordial and primary follicles the oocyte showed distinct amounts of the transcript encoding GHR, in tertiary follicles the mRNA was predominantly localized in the cells of the cumulus oophorus. GHR mRNA was also expressed in the large granulosa lutein cells, in the germinal epithelium, and in the endothelial cells of ovarian vessels. Colocalization of the GHR protein showed a distribution pattern identical to that of the mRNA. In calves, oocyte and follicle cells changed GHR expression in the same way as in the adult ovary. During embryonic development of the ovary, distinct amounts of the mRNA encoding GHR were found in primordial follicles shortly before birth. Our results imply that the GHR is involved in ovarian ontogenesis, especially in early folliculogenesis. PMID- 9746734 TI - Rat prostaglandin D2 synthetase: its tissue distribution, changes during maturation, and regulation in the testis and epididymis. AB - The changes in glutathione-independent prostaglandin D2 synthetase (PGD-S) during maturation in the rat were determined in selected organs by an RIA using PGD-S purified from rat cerebrospinal fluid and a monospecific anti-rat PGD-S polyclonal antibody. In a survey of its tissue distribution in various organ extracts and biological fluids, it was found that the concentration of PGD-S was highest in the epididymis-about 6- and 80-fold greater than that in the brain and testis, respectively. During maturation, PGD-S concentration increased steadily in the testis and epididymis; this is in contrast to the pattern of changes in the brain and liver, which showed a general trend of decline. Reverse transcription-polymerase chain reaction and Southern blotting were used to demonstrate the presence of PGD-S mRNA transcript in the testis and in Sertoli and germ cells. In the epididymis, the steady-state PGD-S mRNA level was highest in the caput, followed by the cauda and corpus. Orchiectomy induced a drastic reduction of PGD-S concentration in all three epididymal compartments. Administration of dihydrotestosterone (DHT) failed to restore the reduced epididymal PGD-S level except in the caput epididymis, where 4 days after DHT treatment the level of PGD-S was restored to about 50% of the pre-orchiectomized level; this suggests that the epididymal PGD-S level is not entirely regulated by androgen and that another yet to be identified testicular factor(s) is likely to be involved in its regulation. Germ cell-conditioned medium was also shown to stimulate PGD-S expression in the Sertoli cell. These results illustrate that PGD S is an important molecule in testicular and epididymal function and that it is likely involved in spermatogenesis and sperm maturation. PMID- 9746735 TI - Isoforms of human recombinant follicle-stimulating hormone: comparison of effects on murine follicle development in vitro. AB - The effects of three isoforms derived from recombinant human FSH on ovarian follicle development in vitro were characterized for the first time. The three subfractions comprised discrete pI ranges of 3. 6-4.6 (acid), 4.5-5.0 (mid), and 5.0-5.6 (least acidic). Follicular growth, estradiol secretion, and antral formation were assessed for each fraction of isoforms in a range of concentrations over a 5-day culture period. Least acidic FSH produced, at and above 1.5 ng/ml, a high percentage of follicles growing above the size threshold necessary for antral formation, whereas mid and acid FSH induced similar growth only at higher concentrations (7.5 ng/ml and 50 ng/ml, respectively). Least acidic FSH specifically induced the most rapid growth of follicles during preantral development. Acid FSH at all concentrations stimulated estradiol-17ss secretion later during culture and antral formation in a lower proportion of follicles than did least acidic and mid FSH. It can be concluded 1) that the least acidic isoform induced fastest preantral growth, producing the largest antral follicles at the lowest dose of all three fractions and 2) that the less and mid acidic isoforms had more impact on stimulation of estradiol production and antral formation than the acid isoform. PMID- 9746736 TI - Thrombospondin-1 expression in human myometrium before and during pregnancy, before and during labor, and in human myometrial cells in culture. AB - The activation of latent transforming growth factor beta (L-TGFbeta) is essential for the action of TGFbeta, which, in turn, is involved in the regulation of expression of some progesterone-responsive genes. One mechanism by which TGFbeta is activated involves thrombospondin (TSP), a protein that binds extracellular proteins. Immunoreactive TSP (irTSP) protein and TSP-1 mRNA in myometrial tissues of ovulatory and pregnant women were localized by immunohistochemistry and in situ hybridization. IrTSP and TSP-1 mRNA were randomly distributed in myometrial smooth muscle cells of some, but not all, tissues of pregnant women at term before labor; but in some areas of most of these tissues, irTSP was intense and commonly localized extracellularly. Intense irTSP and TSP-1 mRNA in myocytes were more common in myometrium during labor. In myometrium from ovulatory women (n = 26), irTSP was localized primarily in vascular smooth muscle cells and was detected occasionally in scattered myocytes. Little TSP-1 mRNA was demonstrable by in situ hybridization in vessels or myocytes of myometrial tissue from ovulatory women (n = 7). By Northern analysis of total RNA, TSP-1 mRNA was detected in myometrial tissue of pregnant women and in human myometrial smooth muscle cells in culture. The levels of TSP-1 mRNA in myometrial tissues of pregnant women during labor (n = 18) were greater than those in myometrium at > 37 wk gestation before labor began (n = 25, p < 0.001). The ratios of TSP-1 to glyceraldehyde 3-phosphate dehydrogenase mRNAs in 3 myometrial tissues during oxytocin-induced labor were not statistically different from those in myometrium during spontaneous labor but were greater than those in myometrium before labor (p < 0.05). The level of TSP-1 mRNA in confluent human myometrial cells in culture was relatively high, was increased by treatment with fetal bovine serum, and was decreased by treatment with platelet-derived growth factor or activators of adenylyl cyclase or protein kinase C. Myometrial cells in culture constitute a useful model for studying the regulation of TSP-1 gene expression in human myometrium. PMID- 9746737 TI - Factors affecting meiotic and developmental competence of primary spermatocyte nuclei injected into mouse oocytes. AB - Mature mouse oocytes that have received the nuclei of pachytene primary spermatocytes (or metaphase I chromosomes of primary spermatocytes) can develop into fertile offspring. However, success rate in this study was low. No more than 3.8% of transferred 2-cell embryos arising from spermatocyte-injected oocytes developed to full term. Nevertheless, the birth of normal offspring seems to suggest that at least in some primary spermatocytes the functional genomic imprinting is complete before transfer and/or consolidated after the transfer. Although injected spermatocyte nuclei could undergo two successive meiotic divisions within oocytes, abnormalities of both divisions were commonly observed, and sister chromatids often separated prematurely during the second meiotic division. Chromosome breakage/rearrangements were also frequently seen before the first cleavage. Such abnormalities of chromosome behavior are probably the major causes of the poor preimplantation development of zygotes arising from primary spermatocyte-injected oocytes. Thus, clinical use of primary spermatocytes as substitutes for spermatozoa in assisted fertilization is not advisable until the causes of chromosomal abnormalities are better understood through extensive animal studies. PMID- 9746738 TI - Changes in expression of contractile FP and relaxatory EP2 receptors in pregnant rat myometrium during late gestation, at labor, and postpartum. AB - Prostaglandins synthesized at parturition may act via specific myometrial receptors as mediators of uterine contractions. Several isoforms of eicosanoid (prostaglandin) receptors, identified by pharmacological means, are linked to contractile (FP, EP1, EP3) or relaxatory (EP2, EP4, IP, DP) intracellular pathways. Changes in mRNA expression of the contractile FP and the relaxatory EP2 receptor were measured in myometrium throughout gestation, at parturition, and postpartum. Timed pregnant rats were killed at 0900 h on Day 16, 18, 20, 21, 21.5, or 22 (parturition) of pregnancy or one day postpartum (n = 5 animals/group). A longitudinal section of myometrium was removed, total RNA was extracted, and semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) was performed for glyceraldehyde phosphate dehydrogenase (GAPDH), calponin, EP2, and FP receptor mRNA expression. Complementary DNA products were run on agarose gels and visualized, and the quantity of cDNA product was measured against a DNA mass ladder. RT-PCR product identity was confirmed by restriction enzyme cleavage. EP2 receptor mRNA expression was highest at Day 16 and declined significantly to Day 21.5 and one day postpartum (p < 0.05, Student-Newman-Keuls procedure). Expression of FP receptor mRNA was low at Day 16 of gestation and increased significantly until delivery (p < 0.05, ANOVA) at Day 22, then fell to prepartum levels at one day postpartum. Myometrial activity at parturition may change from an active quiescent to an active contractile state in concert with a decline in expression of the relaxatory EP2 receptors and up-regulation of contractile FP receptors. PMID- 9746739 TI - Rat oocytes fertilized in modified rat 1-cell embryo culture medium containing a high sodium chloride concentration and bovine serum albumin maintain developmental ability to the blastocyst stage. AB - A suitable chemically defined culture medium for 1-cell rat embryos (mR1ECM) was modified to obtain sperm penetration, and the developmental competence of oocytes fertilized in the medium was compared to that of oocytes fertilized in a traditional medium, modified Krebs-Ringer bicarbonate medium (mKRB). Sperm penetration was not observed when polyvinyl alcohol was replaced with BSA in mR1ECM (mR1ECM-BSA); the incidence was improved only when the osmolarity in mR1ECM-BSA was increased to that in mKRB (310 mOsm) by addition of NaCl. The proportion of oocytes penetrated in mR1ECM-BSA with NaCl increased (71.6 +/- 6.9%), which was not different compared to that in mKRB (76.7 +/- 13.7%). High incidences of sperm penetration (88.8 +/- 4.1% to 93.1 +/- 5.1%) were also observed when NaCl concentration in mR1ECM-BSA was increased from 76.7 mM to 100 130 mM. The incidence of embryos developing to the morula and blastocyst stages was higher when fertilized in mR1ECM-BSA containing 110-130 mM NaCl (91-94%) than in mKRB (70%). A total of 5 offspring were obtained after transfer of the morulae and blastocysts (69 embryos/7 females). These results demonstrate that a high developmental ability of rat embryos to the blastocyst stage is attained when the embryos have been fertilized in mR1ECM-BSA containing 110-130 mM NaCl and then cultured in mR1ECM. PMID- 9746740 TI - Rat epididymal sperm quantity, quality, and transit time after guanethidine induced sympathectomy. AB - Guanethidine, a chemical that selectively abolishes peripheral noradrenergic nerves, was used to investigate the role of sympathetic innervation in the maintenance of epididymal sperm quantity and quality. Four groups of 10 adult male rats each were treated daily for 21 days, by i.p. injections, with either 0 (saline vehicle), 6.25, 12.5, or 25 mg/kg guanethidine. Norepinephrine content was reduced to undetectable levels in the cauda epididymidis in all guanethidine groups after 3 wk of treatment and was reduced to 7.4% of the control values after 1 wk of 6.25 mg/kg treatment. While body weight gain was significantly decreased at 12.5 and 25 mg/kg compared to that in controls, there was a significant increase in the weights of the seminal vesicles/coagulating glands in all treated groups. The number of homogenization-resistant spermatids per testis and the daily sperm production per testis remained unchanged. The weight of the epididymis was significantly increased at 6.25 and 12.5 mg/kg. Moreover, the number of cauda epididymal sperm and the transit time were increased significantly at 6.25 mg/kg (10.2 days) compared to values in the control cauda (6.3 days). Neither serum testosterone levels nor LH was affected in a dosage related manner. There were no effects of guanethidine treatment on cauda epididymal sperm motility or morphology. A quantitative analysis of detergent extracted cauda epididymal sperm proteins by SDS-PAGE revealed no differences, but there were diminutions in seven proteins in homogenates of caput/corpus tissue. Histologic analysis of testis and epididymis sections revealed no differences between control and denervated animals. In a subsequent experiment the lowest effective dosage (6.25 mg/kg) was given to rats for 1 wk, and an increased number of cauda epididymal sperm and a delay in sperm transit were observed. Our results indicate that low-dosage guanethidine exposure denervates the epididymis within 1 wk, thereby delaying epididymal transit; however, neither 1- nor 3-wk exposure produces qualitative changes in the sperm. PMID- 9746741 TI - Fertility of rat epididymal sperm after chemically and surgically induced sympathectomy. AB - Guanethidine, a chemical that selectively blocks sympathetic noradrenergic neurons, was used to investigate the role of sympathetic innervation in the fertility of rat epididymal sperm, using both natural mating and in utero insemination protocols. This animal model correlates, at least in part, with spinal cord injury (SCI) in men. Adult male rats were treated daily by i.p. injections, for 21 or 42 days, with 0 or 6.25 mg/kg guanethidine. To compare the effects of guanethidine-induced sympathectomy with those following surgically induced sympathectomy, the inferior mesenteric ganglion and the proximal hypogastric nerves were removed in another group of rats. Both chemically and surgically induced sympathectomy increased the weight of the epididymis and seminal vesicles/coagulating glands as well as the number and the transit time of cauda epididymal sperm. Neither serum testosterone levels nor LH was affected by treatment with guanethidine. Using natural mating, no litters were produced by guanethidine-treated rats. Chemically denervated rats failed to produce copulatory plugs or ejaculate into the uterus. However, distal cauda epididymal sperm from chemically or surgically denervated rats displayed normal fertilization ability (80%) using in utero inseminations. In addition, the sperm of denervated rats did not show abnormal sperm chromatin structure using an assay that detects DNA damage. We conclude that sympathectomy delays the transit of sperm through the cauda epididymidis and produces ejaculatory dysfunction but does not compromise sperm quality in the distal cauda epididymidis. Moreover, these data provide compelling evidence that there is no association between the prolonged transit time of sperm within the epididymis, i.e., pre-ejaculatory sperm aging, and the fertility of those sperm, which has important implications for artificial insemination using sperm from men with SCI. PMID- 9746742 TI - Beta transforming growth factors (TGFss) at the porcine conceptus-maternal interface. Part I: expression of TGFbeta1, TGFbeta2, and TGFbeta3 messenger ribonucleic acids. AB - Spatial and temporal mRNA expression of beta transforming growth factors (TGFbeta1, TGFbeta2, and TGFbeta3) in porcine uterus and conceptuses was determined during the peri-implantation period (Days 10-14 of gestation). Northern blotting identified a major 3. 5-kilobase (kb) and a minor 2.5-kb transcript for TGFbeta1 mRNA. TGFbeta2 transcripts were 6.2 kb, 5.4 kb, and 2.7 kb, and a single 3. 5-kb transcript was detected for TGFbeta3. With semiquantitative in situ hybridization analysis, progressive increases were detected in TGFbetas 1, 2, and 3 mRNA expression in uterine luminal epithelium (ULE), uterine glands (UGs), and underlying stroma (stroma spongiosum, SS) from Days 10 through 14 of gestation (p < 0.05). In myometrium, TGFbeta mRNA expression did not differ between Days 10 through 14 of gestation. In porcine conceptuses, TGFbetas 1, 2, and 3 mRNA expression was detected in trophectoderm, endoderm, embryonic ectoderm, and mesoderm. For the three TGFbeta isoforms examined, mRNA expression increased 2- to 4-fold in trophectoderm and endoderm between Days 10 and 14 of gestation. TGFbeta1 mRNA levels increase significantly in embryonic ectoderm, but not mesoderm, between Days 12 and 14 of gestation; during that same time, TGFbeta2 mRNA levels increased, but no change was detected in TGFbeta3 mRNA levels, in embryonic ectoderm and mesoderm. Progressive increases in TGFbeta mRNA expression in conceptus trophectoderm, endoderm, ULE, UGs, and SS suggest important roles for these growth factors in porcine conceptus development during the peri-implantation period. PMID- 9746743 TI - Beta transforming growth factors (TGFbeta) at the porcine conceptus-maternal interface. Part II: uterine TGFbeta bioactivity and expression of immunoreactive TGFbetas (TGFbeta1, TGFbeta2, and TGFbeta3) and their receptors (type I and type II). AB - Porcine uterine tissues were collected from Days 10 to 14 of gestation (peri implantation period) or corresponding days of the estrous cycle. Results indicated a marked increase in beta transforming growth factors (TGFbeta1, TGFbeta2, and TGFbeta3) and TGFbeta receptor (type I and type II) immunostaining in uterine luminal epithelium (ULE) between Days 10 and 14 of gestation, but there was no increase in ULE immunostaining on the corresponding days of the estrous cycle. Uterine glands and stroma were intensely immunopositive in pregnant gilts for TGFbeta isoforms and their receptors, but immunostaining was weak to undetectable in cycling gilts. No differences were detected in myometrium, in which immunostaining was moderate in both cycling and pregnant gilts. Additionally, TGFbeta2 and TGFbeta receptor (type I and type II) immunostaining was detected in uterine monocyte/macrophage-like cells. Western blotting detected the presence of all three TGFbeta isoforms in uterine luminal flushings. The CCL64 cell TGFbeta bioassay detected bioactive TGFbetas++ in uterine luminal flushings on Days 12, 13, an 14 of gestation. These results strongly indicate that uterine expression of TGFbetas and their receptors is pregnancy specific and that bioactive TGFbetas are present at the conceptus maternal interface in the peri-implantation period in pigs. Thus TGFbetas are likely to be involved in autocrine-paracrine interactions between the maternal uterus and the conceptus. PMID- 9746744 TI - Sperm and oocyte treatments to improve the formation of male and female pronuclei and subsequent development following intracytoplasmic sperm injection into bovine oocytes. AB - This study assessed pronuclear formation, the chromosomal constitution, and the developmental capacity of bovine zygotes formed by intracytoplasmic injection of oocytes with sperm, treated or not with dithiothreitol (DTT). Oocytes were matured in vitro for 22-24 h and then centrifuged so that sperm, prepared by swim up in the presence or absence of 5 mM DTT, could be injected into the cleared area of the ooplasm. Injected oocytes were activated by treatment with 5 microM ionomycin (5 min) and, after a 3-h interval, with 1.9 mM 6-dimethylaminopurine (DMAP) for 3 h. They were then cocultured with bovine oviductal epithelial cells in M199. Sperm treatment resulted in a significantly higher proportion of male pronucleus formation 16 h after injection (40% vs. 11%; p < 0.0001) and a significantly higher rate of blastocyst development (24% vs. 10%; p < 0.005). Sixty-one percent of blastocysts produced with treated sperm were diploid. Of 12 blastocysts produced with treated sperm and sexed by a polymerase chain reaction, 4 were male and 7 female, and in one a definite diagnosis could not be made. Embryo transfer (2 embryos per heifer) resulted in pregnancies in 6 of 16 recipients at Day 49, but none was carried to term. These results show that the efficiency of bovine intracytoplasmic sperm injection can be improved by sperm pretreatment with DTT and by oocyte activation with ionomycin plus DMAP, although the developmental capacity of the resulting embryos remains limited. PMID- 9746745 TI - Expression of opioid receptors and ligands in pregnant mouse uterus and placenta. AB - The endogenous opioid system has been implicated in the regulation of hormonal secretion, pain perception, and uterine contractility during pregnancy, but there is only limited information about the cellular location of opioid receptor and opioid peptide gene expression in the pregnant rodent uterus and placenta. In this study, we have used in situ hybridization to identify expression sites of mRNAs encoding the delta (delta), kappa (kappa), and mu ( micro) opioid receptors as well as the endogenous opioid peptide precursors proenkephalin (PENK), prodynorphin (PDYN), and proopiomelanocortin (POMC) in pregnant mouse uterus and placenta. Soon after implantation, all three opioid receptor genes as well as POMC and PENK, but not PDYN, were detected in the uterine environment. Each expressed gene exhibited a distinct expression pattern that was generally retained until late gestation. The delta receptor and POMC were coexpressed in the trophoblast giant cells, which remained the only cells of the placenta/uterus to express these two genes throughout gestation. Cells expressing kappa receptors were absent from the placenta but instead were found in the basal part of the decidualized uterine endometrium. While kappa and micro receptors were transiently expressed in the uterine myometrium (until embryonic day 8.5), substantial levels of PENK were continuously detected in this region until at least embryonic day 18. In addition, complementary expression of the micro receptor and PENK genes in the uterus was detected. Taken together, these results suggest multiple roles for the opioid receptors and opioid peptides in maternal adaptation to pregnancy and in supporting embryo growth. PMID- 9746746 TI - Regulation of inducible nitric oxide synthase messenger ribonucleic acid expression in pregnant rat uterus. AB - Nitric oxide synthases catalyze the synthesis of the biomediator, nitric oxide, from L-arginine in a variety of tissues. The expression and regulation of inducible isoform of nitric oxide synthase (NOS II) in the uterus were assessed in this study by reverse transcription-polymerase chain reaction with the use of specific primers. Results showed the following: 1) NOS II mRNA expression in the rat uterus was substantially increased during pregnancy and decreased during labor at term; 2) RU-486 (an antagonist of progesterone) induced preterm labor and was associated with a marked decrease in NOS II mRNA expression to 60.9%, 20.3%, and 2.9% at, respectively, 6, 12, and 24 h after treatment compared with the control value (100%); 3) progesterone administration in pregnant rats significantly increased uterine NOS II gene expression (374.1% vs. 100%); 4) NOS II mRNA in the uterus was significantly reduced by prostaglandin F2alpha (PGF2alpha; 11.6% vs. 100% in control); 5) treatment with progesterone prevented PGF2alpha-induced inhibition in NOS II mRNA expression; 6) ICI 164384, an antiestrogen, significantly increased serum progesterone concentration and stimulated NOS II expression by the uterus in a time-dependent manner; 7) as shown by immunofluorescent studies, cells stained by NOS II antibodies were apparent in the decidual compartment as well as in areas between myometrial cell bundles in the pregnant rat uterus. The density of staining decreased in the specimens at labor and postpartum. We conclude that NOS II gene expression in the rat uterus was enhanced during pregnancy and decreased during labor and postpartum. NOS II in rat uterus is up-regulated by progesterone and down regulated by estrogens and prostaglandins, consistent with their role in uterine activity regulation during pregnancy and labor. PMID- 9746747 TI - Differential expression of Ped gene candidates in preimplantation mouse embryos. AB - The Ped (preimplantation embryonic development) gene influences the rate of mouse preimplantation embryonic development and subsequent survival. Four similar tandem genes in the Q region of the major histocompatibility complex-Q6, Q7, Q8, and Q9-were identified as Ped gene candidates. In this study, expression of these genes during preimplantation development was examined and quantitated by reverse transcription-polymerase chain reaction and single nucleotide primer extension assays in order to investigate their contribution to the Ped gene phenotype. The Q7/Q9 gene pair was found to be transcribed in preimplantation mouse embryos, whereas transcription of the Q6/Q8 gene pair was undetectable. Both Q7 and Q9 are expressed in embryos from one Ped fast strain, C57BL/6, while only the Q9 gene is expressed in another Ped fast strain, B6.K2. These results suggest that both the Q7 and Q9 genes can function as the Ped gene in the mouse. Interestingly, the expression pattern of the Q7 and Q9 genes in preimplantation embryos is the same as in splenic lymphocytes. However, the Q6 and Q8 genes are expressed in splenic lymphocytes but not in preimplantation embryos. Treatment of mouse preimplantation embryos with interferon gamma (gamma-IFN) did not induce expression of the Q6/Q8 genes but enhanced expression of the Q7/Q9 genes. The mechanism of this differential transcription pattern is currently under investigation. PMID- 9746749 TI - Pulsatile luteinizing hormone secretion in the ovariectomized, thyroidectomized red deer hind following treatment with dopaminergic and opioidergic agonists and antagonists. AB - Two experiments were conducted to determine whether dopaminergic or opioidergic pathways are modulated by thyroid gland secretions for seasonal suppression of LH secretion in red deer hinds. Ovariectomized (n = 5) or ovariectomized and thyroidectomized (n = 4) hinds, treated with estradiol implants, received the dopamine agonist bromocriptine or the antagonist sulpiride during pulse bleeds in July (breeding season) and October (nonbreeding season). Comparison of July and October mean plasma LH concentration (3.5 +/- 1.3, 0.7 +/- 0.1 ng/ml, respectively), pulse frequency (1.9 +/- 0.4, 0.7 +/- 0.2 pulses/4 h), and pulse amplitude (1.3 +/- 0.5, 0.7 +/- 0. 02 ng/ml) showed lower (p < 0.05) levels in October, and these levels were not significantly affected by thyroidectomy or drug treatment. In the absence of estradiol implants, the hinds received bromocriptine or morphine during the breeding season (July) and their antagonists, sulpiride or naloxone, respectively, in the nonbreeding season (November). In euthyroid hinds there was a seasonal decrease (p < 0.05) in mean plasma LH concentration, pulse frequency, and pulse amplitude, which did not occur in thyroidectomized hinds. There were no effects of drug treatment on LH concentration except for a small increase following sulpiride in November. Plasma prolactin concentration was significantly increased by antagonists and decreased by agonists on most occasions. We conclude that in red deer hinds, seasonal regulation of LH secretion does not involve dopamine or endogenous opioids and the thyroid gland is required specifically for LH suppression in the absence of estradiol. PMID- 9746748 TI - Expression of steroidogenic acute regulatory protein messenger ribonucleic acid is limited to theca of healthy bovine follicles collected during recruitment, selection, and dominance of follicles of the first follicular wave. AB - Expression of mRNA encoding steroidogenic acute regulatory protein (StAR) in bovine follicles during recruitment and selection was examined. Dairy heifers (4 5/time period) were ovariectomized at 12, 24, 36, 48, 60, 72, 84, or 96 h after initiation of the first follicular wave (Time 0) after estrus. Follicles were collected and stored at -80 degrees C until sectioning. Expression of StAR mRNA was localized by in situ hybridization and quantified by image analysis. Expression of StAR mRNA was first detected in theca interna of antral follicles as small as 0.5 mm in diameter and increased with increasing follicular size (>/= 4 mm; r = 0.75; p < 0.001). StAR mRNA was undetectable in granulosa of healthy follicles at any size or stage of follicular wave examined. However, granulosa or luteinized granulosa of some advanced or late atretic follicles expressed StAR mRNA. During recruitment, StAR mRNA expression in theca cells was similar among recruited follicles (4-8 mm). During selection of dominant follicles (36-48 h), StAR mRNA was expressed in theca of more than one follicle (7-9 mm); therefore, expression of StAR mRNA may not be associated with dominant follicle selection. StAR mRNA in theca was higher (p < 0.05) at 48 h after initiation of the first follicular wave than at 12, 24, and 36 h, and it remained elevated thereafter through 96 h. Dominant follicles expressed more (p < 0.01) StAR mRNA in theca than did subordinate healthy follicles. Healthy follicles expressed higher (p < 0.05) StAR mRNA in theca than atretic follicles. In summary, levels of StAR mRNA increased in theca with stage of follicular wave and size of follicles. Follicular atresia was associated with reduced expression of StAR mRNA in theca cells. The results indicate that expression of StAR mRNA in theca may not be the primary limiting factor for follicular recruitment and selection. PMID- 9746750 TI - Transient expression of a translation initiation factor is conservatively associated with embryonic gene activation in murine and bovine embryos. AB - In the present study the abundance of mRNAs for eukaryotic translation initiation factors eIF-1A (formerly known as eIF-4C), -2alpha, -4A, -4E, and -5 was examined in in vivo-derived mouse embryos throughout preimplantation development using a semiquantitative reverse transcription-polymerase chain reaction assay. Although the mRNA profile for each gene is unique, only mRNA for eIF-1A transiently increases during embryonic gene activation (EGA) at the 2-cell stage, and this was confirmed by an independent hybridization-based assay. In in vitro-developed bovine embryos, mRNA for eIF-1A was transiently detected at the 8-cell stage, when the major activation of the genome occurs in this species. As in the mouse, detection in 8-cell bovine embryos was sensitive to the transcriptional inhibitor alpha-amanitin. It was also observed at the same time relative to cleavage in embryos cultured in defined medium under a reduced oxygen environment, and in medium supplemented with serum and somatic cells in 5% CO2 in air. Neither the chronology of early cleavage divisions nor the yield of bovine blastocysts differed in these culture media. Our results suggest that transient expression of eIF-1A in the mouse and cow is a conserved pattern of gene expression associated with EGA in mammals. PMID- 9746751 TI - Positive association between expression of follicle-stimulating hormone beta and activin betaB-subunit genes in boars. AB - This study tested our hypothesis that inhibin/activin (I/A) betaB subunit and not follistatin (FS) gene expression relates positively to plasma FSH concentrations in the anterior pituitary gland of boars. Mature crossbred boars (n = 12) were selected for divergence in plasma FSH concentrations, and their anterior pituitary glands were evaluated for expression of the FSHbeta, I/A ssB, FS, calmodulin, and GnRH receptor (GnRH-R) genes by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) and/or RNase protection assays (RPAs). Expression of I/A ssB was greater (p < 0. 01) in the six boars with high FSH than in the six with low FSH; expression of the I/A betaB-subunit gene was positively correlated to that of the FSHbeta gene (RT-PCR: r = 0.96; p < 0.01; RPA: r = 0.68; p < 0.05). In contrast, expression of the FS (p > 0.10), GnRH-R (p > 0. 08), and calmodulin (p > 0.10) genes was similar in the two groups of boars. Additionally, expression of the FSHbeta gene was correlated positively with pituitary and plasma FSH concentrations (r = 0.69 and 0.88, respectively; p < 0.05). These results support the hypothesis that activin B is partially responsible for elevated FSH concentrations in boars. Furthermore, the expression difference of the calmodulin gene observed previously between Meishan and White Composite boars represents a breed difference unrelated to FSH. PMID- 9746752 TI - Ontogeny of stem cell factor receptor (c-kit) messenger ribonucleic acid in the ovine corpus luteum. AB - Stem cell factor (SCF) is a pleiotropic growth factor that is expressed by the ovine corpus luteum throughout its life span by both small and large steroidogenic cells. Determination of the action of SCF, however, requires localization of its receptor, c-kit; therefore, the objectives of the present study were to identify and localize c-kit within corpora lutea. Two cDNAs encoding different portions of the c-kit molecule were amplified by the polymerase chain reaction. The first was a 558-base pair (bp) cDNA encoding portions of the transmembrane and tyrosine kinase domains; the second was a 632 bp cDNA encoding most of the ligand-binding domain. Expression of c-kit was quantified by RNase protection assay of total cellular RNA collected on Days 3, 7, 10, 13, and 16 (n = 4, 4, 5, 4, and 4 per group, respectively) of the estrous cycle (Day 0 = estrus). The level of c-kit mRNA was low early in the luteal phase, reached (p < 0.05) maximum levels on Day 13, and then decreased (p < 0.01) on Day 16. On Day 3 (n = 4), c-kit was expressed in a cell-specific manner throughout the corpus luteum; identity of the specific cell types expressing c kit could not be determined at this stage. On Day 14 (n = 4), c-kit did not appear to be expressed within large luteal cells but was prominently expressed in cells that surrounded large luteal cells and that possessed the morphological characteristics of small luteal cells and endothelial cells. Given the temporal regulation of c-kit expression within the corpus luteum, these data suggest that luteal SCF may act locally. PMID- 9746753 TI - Identification of specific relaxin-binding cells in the human female. AB - Relaxin is secreted during pregnancy, but it has no verified effects in humans. The objective of the present study was to identify the cells containing specific relaxin-binding sites in the uterine cervix, vagina, uterus, mammary glands, mammary nipples, and term placenta in the human. The uterine cervix, vagina, and uterus were obtained from hysterectomy specimens. Mammary glands and nipples were obtained after modified radical mastectomy. Placenta was obtained after normal delivery. Tissue samples were cut into slices (0.5-3 cm3), frozen in liquid nitrogen, and cryosectioned (8 microm). Cells that bind relaxin were identified by sequential application of biotinylated porcine relaxin probe, antibiotin immunoglobulin G conjugated to 1 nm colloidal gold, and silver enhancement for signal amplification. Relaxin bound with specificity to epithelial cells, smooth muscle cells, and blood vessels in the cervix, vagina, uterus, and mammary nipples; to epithelial cells and blood vessels in the mammary glands; and to skin of the mammary nipples. In addition, relaxin bound to individual cell types within the term placenta (amnion epithelium, syncytiotrophoblasts, blood vessels), and to sebaceous glands within the nipples. We conclude that the specific relaxin-binding cells probably contain relaxin receptors. Identification of putative relaxin receptors may provide insight into physiological and/or therapeutic roles of relaxin in the human. PMID- 9746754 TI - Calcium homeostasis in early hamster preimplantation embryos. AB - The development in culture of 1-cell hamster embryos prior to the completion of fertilization is not well understood. In this study it was observed that culture for only 6 h of these early 1-cell embryos collected before pronuclei formation (3 h post-egg activation; PEA) significantly increased intracellular free calcium levels (194.3 +/- 3.1 nM) compared to levels in similarly aged 1-cell embryos collected from the oviduct at 9 h PEA, after pronuclei formation is complete (134.2 +/- 6.8 nM). Not only was the developmental competence of cultured 3-h PEA embryos with elevated intracellular free calcium levels compromised as compared with that of embryos collected from the oviduct at 9 h PEA; these embryos also had impaired cytoplasmic mitochondrial distribution (ratio of 0.62 +/- 0. 06 for cultured embryos compared to 0.44 +/- 0.04 for in vivo-developed embryos) and decreased lactate metabolism (2.93 +/- 0. 22 pmol/embryo per 3 h for cultured embryos compared to 5.37 +/- 0. 36 for in vivo-developed embryos). This impairment in mitochondrial distribution and function and reduced development in culture by 3-h PEA embryos appears related to the ability to regulate intracellular calcium homeostasis. Intracellular free calcium levels were reduced by culture with increased medium magnesium concentrations, calcium channel inhibitors nifedipine or verapamil, or an intracellular calcium chelator. All of these treatments also stimulated development of 3-h PEA embryos to the morula/blastocyst stages and prevented impairment in mitochondrial organization and function. Conversely, culture with low medium magnesium and high calcium concentrations that increased intracellular free calcium levels resulted in low development and reduced mitochondrial function. Therefore, it appears that removal of the early embryo from the oviduct results in an inability to regulate intracellular calcium levels. As increased magnesium concentrations, nifedipine, and verapamil inhibit L-gated calcium channels, it may be a loss of regulation of these channels that alters calcium homeostasis resulting in impaired developmental competence. PMID- 9746756 TI - gamma-Chain dysfibrinogenemias: molecular structure-function relationships of naturally occurring mutations in the gamma chain of human fibrinogen. PMID- 9746755 TI - Estrogen and lipopolysaccharide stimulation of prostacyclin production and the levels of cyclooxygenase and nitric oxide synthase in ovine uterine arteries. AB - Several enzymes play a role in vasodilation, including cyclooxygenase, which converts arachidonic acid into prostaglandins, and nitric oxide synthase, which converts arginine to citrulline and yields nitric oxide. The effects of endogenous and exogenous estrogen and lipopolysaccharide on uterine artery production of prostacyclin, and levels of cyclooxygenase and nitric oxide synthase were examined. Uterine arteries collected from ewes during the follicular (Day -1 to 0, Day 0 = estrus) or luteal (Day 10) phase were treated in vitro with lipopolysaccharide. In addition, ovariectomized ewes were treated in vivo with estradiol-17beta (5 microg/kg; 120 min) or a vehicle control; arteries from the uteri were treated in vitro with lipopolysaccharide. After 24 h of lipopolysaccharide treatment, culture media were collected for measurement of 6 keto-prostaglandin F1alpha (the stable metabolite of prostacyclin). These uterine arteries were homogenized, and the level of cyclooxygenase and nitric oxide synthase was determined by Western analysis. Lipopolysaccharide stimulated (p < 0.02) prostacyclin production by uterine arteries from both follicular- and luteal-phase sheep although phase of the estrous cycle did not affect prostacyclin responses (p = 0.56) to lipopolysaccharide. In contrast, uterine arteries from ovariectomized sheep treated with estradiol-17beta produced more prostacyclin (p < 0.001) in response to lipopolysaccharide than did uterine arteries from ovariectomized sheep treated with the vehicle control. There was no effect of phase (follicular or luteal) of the estrous cycle on either cyclooxygenase-1 or -2 gene expression. Lipopolysaccharide increased (p = 0.0002) gene expression of cyclooxygenase-2, but not cyclooxygenase-1, in both follicular and luteal-phase ewes, which was significantly correlated (r2 = 0.91, p = 0.003) with uterine artery production of prostacyclin. Uterine arteries from follicular phase sheep expressed significantly more nitric oxide synthase-III after lipopolysaccharide exposure than did uterine arteries from luteal-phase ewes (p = 0.03). In contrast, nitric oxide synthase-II was not detected in uterine arteries after lipopolysaccharide exposure. These results suggest that estrogen plays a role in regulating uterine artery responses to lipopolysaccharide. PMID- 9746757 TI - DNA-Dependent protein kinase activity correlates with clinical and in vitro sensitivity of chronic lymphocytic leukemia lymphocytes to nitrogen mustards. AB - The objective of this study is to investigate the role of DNA-dependent protein kinase (DNA-PK) in the chronic lymphocytic leukemia (CLL) lymphocyte response to nitrogen mustard therapy. DNA-PK is a nuclear serine/threonine kinase that functions in DNA double-strand break repair and in the joining process in recombination mechanisms. In a series of 34 patients with B-CLL, either untreated (n = 16) or resistant to chlorambucil (n = 18), the kinase activity of the complex, as determined by its capacity to phosphorylate a peptide substrate in vitro, is increased in the resistant samples as compared with the untreated ones (24.4 +/- 2.6 arbitrary units [a.u.] [range, 12.7 to 55.8 a.u.] versus 8.1 +/- 2.8 a.u. [range, 0.9 to 44.5 a.u.], respectively (P < .0001]), independent of other clinical and biological factors. Linear regression analysis shows an excellent correlation between the level of DNA-PK activity and the inherent in vitro sensitivity of CLL lymphocytes to chlorambucil (r = .875, P =.0001). The regulation of DNA-PK activity was associated with increased DNA-binding activity of its regulatory subunit, the Ku heterodimer, in resistant samples. These results suggest that this activity is a determinant in the cellular response to chlorambucil and participates in the development of nitrogen mustard-resistant disease. The increase in DNA-PK activity might contribute to the enhanced cross link repair that we previously postulated to be a primary mechanism of resistance to nitrogen mustards in CLL. PMID- 9746758 TI - Expression status of BCL-6 and syndecan-1 identifies distinct histogenetic subtypes of Hodgkin's disease. AB - The tumor cells in most cases of Hodgkin's disease (HD) have been recently recognized to originate from the B-cell lineage, but their precise differentiation stage is not fully clarified. Recently, we have reported that the histogenesis of B-cell lymphomas may be assessed by monitoring the expression pattern of BCL-6, a transcription factor expressed in germinal center (GC) B cells, and CD138/syndecan-1 (syn-1), a proteoglycan associated with post-GC, terminal B-cell differentiation. In this study, we have applied these two markers to the study of HD histogenesis. We have found that in nodular lymphocyte predominance HD (NLPHD) tumor cells consistently display the BCL-6(+)/syn-1(-) phenotype, indicating their derivation from GC B cells. Conversely, classic HD (CHD) is heterogeneous because the tumor cells of a fraction of CHD display the BCL-6(-)/syn-1(+) phenotype of post-GC B-cells, whereas another fraction of CHD is constituted by a mixture of tumor cells reflecting the GC (BCL-6(+)/syn-1(-)) or post-GC (BCL-6(-)/syn-1(+)) phenotypes. BCL-6(-)/syn-1(+) tumor cells of CHD are mostly found surrounded by T cells expressing CD40L, consistent with the observation that CD40 signaling downregulates BCL-6 expression. These data indicate that tumor cells of NLPHD uniformly display a GC B-cell phenotype, whereas the phenotype of tumor cells of CHD appears to be modulated by the surrounding cellular background, particularly CD40L+ reactive T cells. PMID- 9746759 TI - The Fanconi anemia proteins FAA and FAC function in different cellular compartments to protect against cross-linking agent cytotoxicity. AB - Fanconi anemia (FA) is an autosomal recessive disease characterized by chromosomal instability, bone marrow failure, and a high risk of developing malignancies. Although the disorder is genetically heterogeneous, all FA cells are defined by their sensitivity to the apoptosis-inducing effect of cross linking agents, such as mitomycin C (MMC). The cloned FA disease genes, FAC and FAA, encode proteins with no homology to each other or to any known protein. We generated a highly specific antibody against FAA and found the protein in both the cytoplasm and nucleus of mammalian cells. By subcellular fractionation, FAA is also associated with intracellular membranes. To identify the subcellular compartment that is relevant for FAA activity, we appended nuclear export and nuclear localization signals to the carboxy terminus of FAA and enriched its localization in either the cytoplasm or the nucleus. Nuclear localization of FAA was both necessary and sufficient to correct MMC sensitivity in FA-A cells. In addition, we found no evidence for an interaction between FAA and FAC either in vivo or in vitro. Together with a previous finding that FAC is active in the cytoplasm but not in the nucleus, our results indicate that FAA and FAC function in separate subcellular compartments. Thus, FAA and FAC, if functionally linked, are more likely to be in a linear pathway rather than form a macromolecular complex to protect against cross-linker cytotoxicity. PMID- 9746760 TI - A novel mutation in the coding sequence of the FY*B allele of the Duffy chemokine receptor gene is associated with an altered erythrocyte phenotype. AB - The Duffy blood group system is of clinical and biological significance. Antibodies to Duffy antigens are responsible for some cases of transfusion incompatibility and newborn hemolytic disease. The Duffy protein is a receptor for the Plasmodium vivax erythrocyte-binding protein and is also a receptor for various chemokines (thus renamed Duffy Antigen Receptor for Chemokines [DARC]). The two Duffy polymorphic antigens, Fya and Fyb (coded by the FY*A and FY*B alleles), are present on erythrocyte membranes. The Fy(a-b-) phenotype is the predominant one in populations of black people and also occurs in other populations, including some non-Ashkenazi Jewish groups. The Fy(a-b-) phenotype has been associated with a mutation in the FY*B promoter at the GATA box that abolishes the expression of erythrocyte Duffy protein. We describe here a novel mutation, present in the FY*B coding sequence (271C --> T), that is associated with some Fy(b-) phenotypes among non-Ashkenazi Jews and among Brazilian blacks. The mutation is present in Fy(b-) individuals, who have wild-type FY*B GATA and carry the previously described 304G --> A substitution. The 271C --> T and 304G - > A can be identified by restriction enzyme-generated restriction fragment length polymorphisms. The 271C --> T substitution represents a considerable change in chemical nature (Arg91 --> Cys), one which may affect the antigenic determinants of DARC, and thus be of clinical significance. The mutation may have implications for some physiological roles of DARC and be of interest in malaria research and in studies of population genetics. PMID- 9746761 TI - Caspases mediate retinoic acid-induced degradation of the acute promyelocytic leukemia PML/RARalpha fusion protein. AB - All-trans-retinoic acid (RA) treatment induces morphological remission in acute promyelocytic leukemia (APL) patients carrying the t(15;17) and expressing the PML/RARalpha product by inducing terminal differentiation of the leukemic clone. RA treatment induces downregulation of PML/RARalpha and reorganization of the PML nuclear bodies. These events have been proposed to be essential for the induction of APL cell differentiation by RA. Here, we show that in the APL-derived NB4 cell line as well as in myeloid precursor U937 cells expressing the PML/RARalpha (U937/PR9) and in blasts from APL patients, the PML/RARalpha fusion protein is cleaved by a caspase 3-like activity induced by RA treatment. In fact, a caspase 3-like activity is detectable in PML/RARalpha expressing cells after RA treatment, and selective caspase inhibitor peptides are able to prevent the RA induced degradation of the fusion protein in vivo and in vitro. Using recombinant caspases and PML/RARalpha deletion mutants we mapped a caspase 3 cleavage site (Asp 522) within the alpha-helix region of the PML component of the fusion protein. The extent of PML/RARalpha cleavage directly correlates with the ability of RA to restore the normal PML nuclear bodies (NBs) pattern. However, RA-induced differentiation is not prevented by the persistence of the fusion product and occurs in the absence of normally structured PML NBs. These results indicate that PML/RARalpha is directly involved in conferring RA sensitivity of APL cells and that the RA-induced reassembly of PML NBs is the consequence of the disappearance of PML/RARalpha. PMID- 9746762 TI - Deficient major histocompatibility complex class II antigen presentation in a subset of Hodgkin's disease tumor cells. AB - Hodgkin's disease is a common malignancy of the lymphoid system. Although the scarce Hodgkin and Reed-Sternberg (HRS) tumor cells in involved tissue synthesize major histocompatibility complex (MHC) class II and costimulatory molecules such as CD40 or CD86, it is unclear whether these tumor cells are operational antigen presenting cells (APC). We developed an immunofluorescence-based assay to determine the number of MHC class II molecules present on the surface of single living HRS cells. We found that in fresh Hodgkin's disease lymph node biopsies, a subset of HRS cells express a substantial number of surface MHC class II molecules that are occupied by MHC class II-associated invariant chain peptides (CLIP), indicating deficient loading of MHC class II molecules with antigenic peptides. Cultured Hodgkin's disease-derived (HD) cell lines, however, were found to express few MHC class II molecules carrying CLIP peptides on the cell surface and were shown to generate sodium dodecyl sulphate (SDS)-stable MHC class II alphabeta dimers. In addition to showing deficient MHC class II antigen presentation in a subset of HRS cells, our results show that the widely used HD cell lines are not ideal in vitro models for the disease. The disruption of MHC class II-restricted antigen presentation in HRS cells could represent a key mechanism by which these tumor cells escape immune surveillance. PMID- 9746764 TI - Transduction of murine bone marrow cells with an MDR1 vector enables ex vivo stem cell expansion, but these expanded grafts cause a myeloproliferative syndrome in transplanted mice. AB - Attempts to expand repopulating hematopoietic cells ex vivo have yielded only modest amplification in stem cell numbers. We now report that expression of an exogenous human multi-drug resistance 1 (MDR1) gene enables dramatic ex vivo stem cell expansion in the presence of early acting hematopoietic cytokines. Bone marrow cells were transduced with retroviral vectors expressing either the MDR1 gene or a variant of human dihydrofolate reductase (DHFR), and then expanded for 12 days in the presence of interleukin-3 (IL-3), IL-6, and stem cell factor. When these cells were injected into nonirradiated mice, high levels of long-term engraftment were only seen with MDR1-transduced grafts. To verify that expansion of MDR1-transduced repopulating cells had occurred, competitive repopulation assays were performed using MDR1 expanded grafts. These experiments showed progressive expansion of MDR1-transduced repopulating cells over the expansion period, with a 13-fold overall increase in stem cells after 12 days. In all of the experiments, mice transplanted with expanded MDR1-transduced stem cells developed a myeloproliferative disorder characterized by high peripheral white blood cell counts and splenomegaly. These results show that MDR1-transduced stem cells can be expanded in vitro using hematopoietic cytokines without any drug selection, but enforced stem cell self-renewal divisions can have adverse consequences. PMID- 9746763 TI - Induction of endothelial PAS domain protein-1 by hypoxia: characterization and comparison with hypoxia-inducible factor-1alpha. AB - Hypoxia results in adaptive changes in the transcription of a range of genes including erythropoietin. An important mediator is hypoxia-inducible factor-1 (HIF-1), a DNA binding complex shown to contain at least two basic helix-loop helix PAS-domain (bHLH-PAS) proteins, HIF-1alpha and aryl hydrocarbon nuclear receptor translocator (ARNT). In response to hypoxia, HIF-1alpha is activated and accumulates rapidly in the cell. Endothelial PAS domain protein 1 (EPAS-1) is a recently identified bHLH-PAS protein with 48% identity to HIF-1alpha, raising the question of its role in responses to hypoxia. We developed specific antibodies and studied expression and regulation of EPAS-1 mRNA and protein across a range of human cell lines. EPAS-1 was widely expressed, and strongly induced by hypoxia at the level of protein but not mRNA. Comparison of the effect of a range of activating and inhibitory stimuli showed striking similarities in the EPAS-1 and HIF-1alpha responses. Although major differences were observed in the abundance of EPAS-1 and HIF-1alpha in different cell types, differences in the inducible response were subtle with EPAS-1 protein being slightly more evident in normoxic and mildly hypoxic cells. Functional studies in a mutant cell line (Ka13) expressing neither HIF-1alpha nor EPAS-1 confirmed that both proteins interact with hypoxically responsive targets, but suggest target specificity with greater EPAS-1 transactivation (relative to HIF-1alpha transactivation) of the VEGF promoter than the LDH-A promoter. PMID- 9746765 TI - The natural history of fetomaternal alloimmunization to the platelet-specific antigen HPA-1a (PlA1, Zwa) as determined by antenatal screening. AB - Immunization against the human platelet antigen (HPA)-1 alloantigen is the most common cause of severe fetal and neonatal thrombocytopenia. Fetal therapy has substantial risks and its indications need better definition. Of 24,417 consecutive pregnant women, 618 (2.5%) were HPA-1a negative of whom 385 entered an observational study. All were HLA-DRB3*0101 genotyped and screened for anti HPA-1a. Their partners and neonates were HPA-1 genotyped and the latter were assessed by cord blood platelet counts and cerebral ultrasound scans. Anti-HPA-1a was detected in 46 of 387 pregnancies (12.0%; 95% CI 8.7%-15.2%). All but one were HLA-DRB3*0101 positive (odds ratio 140; 95% CI 19-1035; P< .00001). One baby died in utero, and of 26 HPA-1a-positive babies born to women with persistent antenatal antibodies, 9 were severely thrombocytopenic (8 with a count <10 x 10(9)/L, 1 with a large porencephalic cyst), 10 were mildly thrombocytopenic, whereas 7 had normal platelet counts. Severe thrombocytopenia was significantly associated with a third trimester anti-HPA-1a titer >/= 1:32 (P = . 004), but was not observed in babies of women with either transient or postnatal-only antibodies. HPA-1a alloimmunization complicates 1 in 350 unselected pregnancies, resulting in severe thrombocytopenia in 1:1,200. HPA-1a and HLA-DRB3*0101 typing combined with anti-HPA-1a titration allows selection of the majority of pregnancies at risk of severe thrombocytopenia. PMID- 9746766 TI - Early treatment of acute graft-versus-host disease with high- or low-dose 6 methylprednisolone: a multicenter randomized trial from the Italian Group for Bone Marrow Transplantation. AB - Ninety-five patients undergoing an allogeneic bone marrow transplant (BMT) and developing acute graft-versus-host disease (aGvHD) were randomized to receive low dose intravenous 6-methylprednisolone (6MPred; 2 mg/kg /d; n = 47) or high-dose 6MPred (10 mg/kg/d; n = 48) for 5 days, with subsequent tapering doses. On day 5 patients not responding or progressing on low-dose 6MPred could be switched to high-dose 6MPred. All patients, aged 1 to 55 years, were recipients of unmanipulated BMT from HLA identical sibling donors. Patients were stratified at randomization for age (/= 20 years), disease (acute leukemia, chronic myeloid leukemia [CML], nonneoplastic disease), disease status (early/advanced), and GvHD prophylaxis (cyclosporin/cyclosporin + methotrexate). Primary endpoints were response to treatment and evolution of aGvHD to grade III-IV. Secondary endpoints were cytomegalovirus (CMV) infections, transplant-related mortality (TRM), and relapse. The median interval between BMT and treatment was 12 days (6 to 43). Results in the two groups (2 v 10 mg/kg) were as follows: response of aGvHD 68% versus 71% (P = .9), evolution to aGvHD grade III-IV 17% versus 20% (P = . 6), CMV infections 55% versus 60% (P = .7), 3-year actuarial TRM 28% versus 32% (P = .7), relapse 17% versus 7% (P = .1). The actuarial survival at 3 years was 63% versus 62% (P = .9) with a median follow up of 580 and 778 days. On day 5 of therapy, 26 patients assigned to low-dose (2 mg/kg) 6MPred were switched to a higher dose of 6MPred because of no response or progression. Their actuarial TRM was 46%, which is significantly higher than TRM of patients who responded on 2 mg/kg and continued with tapering doses (TRM = 16%, P = .007). In conclusion, early treatment of acute GvHD with 6MPred 10 mg/kg/d does not improve the response rate as compared with 2 mg/kg/d, nor does it prevent evolution to aGvHD grade III-IV. CMV infections, TRM, and survival were also comparable. A group of patients at high risk of TRM can be identified after 5 days of treatment with 6MPred 2 mg/kg and could be eligible for alternative forms of therapy. PMID- 9746767 TI - BCL-6 gene mutations in posttransplantation lymphoproliferative disorders predict response to therapy and clinical outcome. AB - Posttransplantation lymphoproliferative disorders (PT-LPDs) represent a heterogeneous group of Epstein-Barr virus-associated lymphoid proliferations that arise in immunosuppressed transplant recipients. Some of these lesions regress after a reduction in immunosuppressive therapy, whereas some progress despite aggressive therapy. Morphological, immunophenotypic, and immunogenotypic criteria have not been useful in predicting clinical outcome. Although structural alterations in oncogenes and/or tumor suppressor genes identified in some PT-LPDs correlate with a poor clinical outcome, the presence of these alterations has not been a consistently useful predictor of lesion regression after reduction of immunosuppression. We examined 57 PT-LPD lesions obtained from 36 solid organ transplant recipients for the presence of mutations in the BCL-6 proto-oncogene using single-strand conformation polymorphism and sequence analysis, followed by correlation with histopathologic classification and clinical outcome, which was known in 33 patients. BCL-6 gene mutations were identified in 44% of the specimens and in 44% of the patients; none were identified in the cases classified as plasmacytic hyperplasia. However, mutations were present in 43% of the polymorphic lesions and 90% of the PT-LPDs diagnosed as non-Hodgkin's lymphoma or multiple myeloma. BCL-6 gene mutations predicted shorter survival and refractoriness to reduced immunosuppression and/or surgical excision. Our results suggest that the BCL-6 gene structure is a reliable indicator for the division of PT-LPDs into the biological categories of hyperplasia and malignant lymphoma, of which only the former can regress on immune reconstitution. The presence of BCL-6 gene mutations may be a useful clinical marker to determine whether reduction in immunosuppression should be attempted or more aggressive therapy should be instituted. PMID- 9746768 TI - Phase III study comparing methotrexate and tacrolimus (prograf, FK506) with methotrexate and cyclosporine for graft-versus-host disease prophylaxis after HLA identical sibling bone marrow transplantation. AB - We report the results of a phase III open-label, randomized, multicenter trial comparing tacrolimus/methotrexate to cyclosporine/methotrexate for graft-versus host disease (GVHD) prophylaxis after HLA-identical sibling marrow transplantation in patients with hematologic malignancy. The primary objective of this study was to compare the incidence of moderate to severe (grade II-IV) acute GVHD. Secondary objectives were to compare the relapse rate, disease-free survival, overall survival, and the incidence of chronic GVHD. Patients were stratified according to age (<40 v >/=40) and for male recipients of a marrow graft from an alloimmunized female. There was a significantly greater proportion of patients with advanced disease randomized to tacrolimus arm (P = . 02). The incidence of grade II-IV acute GVHD was significantly lower in patients who received tacrolimus than patients in the cyclosporine group (31.9% and 44.4%, respectively; P = .01). The incidence of grade III-IV acute GVHD was similar, 17.1% in cyclosporine group and 13.3% in the tacrolimus group. There was no difference in the incidence of chronic GVHD between the tacrolimus and the cyclosporine group (55.9% and 49.4%, respectively; P = .8). However, there was a significantly higher proportion of patients in the cyclosporine group who had clinical extensive chronic GVHD (P = . 03). The relapse rates of the two groups were similar. The patients in the cyclosporine arm had a significantly better 2 year disease-free survival and overall survival than patients in the tacrolimus arm, 50.4% versus 40.5% (P = .01) and 57.2% versus 46.9% (P = .02), respectively. The significant difference in the overall and disease-free survival was largely the result of the patients with advanced disease, 24.8% with tacrolimus versus 41.7% with cyclosporine (P = .006) and 20.4% with tacrolimus versus 28% with cyclosporine (P = .007), respectively. There was a higher frequency of deaths from regimen-related toxicity in patients with advanced disease who received tacrolimus. There was no difference in the disease-free and overall survival in patients with nonadvanced disease. These results show the superiority of tacrolimus/methotrexate over cyclosporine/methotrexate in the prevention of grade II-IV acute GVHD with no difference in disease-free or overall survival in patients with nonadvanced disease. The survival disadvantage in advanced disease patients receiving tacrolimus warrants further investigation. PMID- 9746769 TI - A special fluorescent in situ hybridization technique to study peripheral blood and assess the effectiveness of interferon therapy in chronic myeloid leukemia. AB - Using a highly sensitive fluorescence in situ hybridization method with probes for BCR and ABL1 (D-FISH), we studied 37 paired sets of bone marrow and blood specimens, collected within 24 to 96 hours of each other, from 10 patients before and during treatment for chronic myeloid leukemia (CML). The normal range for 500 interphase nuclei was 1.5 hours), it was composed of ferritin and LMW-Fe. The kinetics of iron release were identical for HH monocytes. A high percentage of the total amount of iron was released as Hb both by viable normal and HH monocytes, suggesting that iron release as Hb is a physiologic process, which may occur whenever the erythrocyte-processing capacity of macrophages is exceeded. Most remarkably, HH monocytes released twice as much iron in a LMW form as control cells. Iron released in the form of LMW-Fe readily binds to plasma transferrin and may contribute to the high transferrin saturation and the occurrence of circulating nontransferrin-bound iron observed in HH patients. PMID- 9746793 TI - Transgenic mice expressing human fetal globin are protected from malaria by a novel mechanism. AB - Studies in vitro by Pasvol et al (Nature, 270:171, 1977) have indicated that the growth of Plasmodium falciparum in cells containing fetal hemoglobin (HbF = alpha2gamma2) is retarded, but invasion is increased, at least in newborn cells. Normal neonates switch from about 80% HbF at birth to a few percent at the end of the first year of life. Carriers of beta-thalassemia trait exhibit a delay in the normal HbF switch-off, which might partially explain the protection observed in populations with this gene. To study this hypothesis in vivo, we used transgenic (gamma) mice expressing human Agamma and Ggamma chains resulting in 40% to 60% alpha2Mgamma2 hemoglobin, infected with rodent malaria. Two species of rodent malaria were studied. P chabaudi adami causes a nonlethal infection, mainly in mature red blood cells (RBC). P yoelii 17XNL is a nonlethal infection, invading primarily reticulocytes, whereas P yoelii 17XL is a lethal variant of P yoelii 17XNL and causes death of mice in approximately 1 to 2 weeks. Data indicate that this strain may cause a syndrome resembling cerebral malaria caused by P falciparum (Am J Trop Med Hyg, 50:512, 1994). In gamma transgenic mice infected with P chabaudi adami, the parasitemia rose more quickly (in agreement with Pasvol) than in control mice, but was cleared more rapidly. In mice infected with P yoelii 17XNL, a clear reduction in parasitemia was observed. Interestingly, splenectomy before this infection, did not reverse protection. The most striking effect was in lethal P yoelii 17XL infection. Control mice died between 11 to 13 days, whereas gamma mice cleared the infection by day 22 and survived, a phenomenon also observed in splenectomized animals. These results suggest that HbF does indeed have a protective effect in vivo, which is not mediated by the spleen. In terms of mechanisms, light microscopy showed that intraerythrocytic parasites develop slowly in HbF erythrocytes, and electron microscopy showed that hemozoin formation was defective in transgenic mice. Finally, digestion studies of HbF by recombinant plasmepsin II demonstrated that HbF is digested only half as well as hemoglobin A (HbA). We conclude that HbF provides protection from P falciparum malaria by the retardation of parasite growth. The mechanism involves resistance to digestion by malarial hemoglobinases based on the data presented and with the well-known properties of HbF as a super stable tetramer. In addition, the resistance of normal neonates for malaria can now be explained by a double mechanism: increased malaria invasion rates, reported in neonatal RBC, will direct parasites to fetal cells, as well as F cells, and less to the approximately 20% of HbA containing RBC, amplifying the antimalarial effects of HbF. PMID- 9746794 TI - Early phagocytosis of glucose-6-phosphate dehydrogenase (G6PD)-deficient erythrocytes parasitized by Plasmodium falciparum may explain malaria protection in G6PD deficiency. AB - In population-based studies it has been established that inherited deficiency of erythrocyte (E) glucose-6-phosphate dehydrogenase (G6PD) confers protection against severe Plasmodium falciparum (P falciparum) malaria. Impaired growth of parasites in G6PD-deficient E in vitro has been reported in some studies, but not in others. In a systematic analysis, we have found that with five different strains of P falciparum (FCR-3, KI, C10, HB3B, and T9/96), there was no significant difference in either invasion or maturation when the parasites were grown in either normal or G6PD-deficient (Mediterranean variant) E. With all of these strains and at different maturation stages, we were unable to detect any difference in the amount of P falciparum-specific G6PD mRNA in normal versus deficient parasitized E. The rate of 14C-CO2 production from D-[1-14C] glucose (which closely reflects intracellular activity of G6PD) contributed by the parasite was very similar in intact normal and deficient E. By contrast, in studies of phagocytosis of parasitized E by human adherent monocytes, we found that when the parasites were at the ring stage (ring-stage parasitized E [RPE]), deficient RPE were phagocytosed 2.3 times more intensely than normal RPE (P = .001), whereas there was no difference when the parasites were at the more mature trophozoite stage (trophozoite-stage parasitized E [TPE]). Phagocytic removal markers (autologous IgG and complement C3 fragments) were significantly higher in deficient RPE than in normal RPE, while they were very similar in normal and deficient TPE. The level of reduced glutathione was remarkably lower in deficient RPE compared with normal RPE. We conclude that impaired antioxidant defense in deficient RPE may be responsible for membrane damage followed by phagocytosis. Because RPE, unlike TPE, are nontoxic to phagocytes, the increased removal by phagocytosis of RPE would reduce maturation to TPE and to schizonts and may be a highly efficient mechanism of malaria resistance in deficient subjects. PMID- 9746795 TI - Rh-deficiency of the regulator type caused by splicing mutations in the human RH50 gene. AB - The Rh polypeptides and the glycoproteins Rh50, CD47, LW, and glycophorin B, which interact in the red blood cell membrane to form a multisubunit complex, are lacking or are severely reduced in the Rh-deficiency syndrome. We previously reported that in several Rhnull patients the RH50 gene was altered at the coding sequence level, resulting in either a single amino acid substitution or the synthesis of a truncated polypeptide. In the present report, we have detected two mutations in the intronic region of the RH50 gene that identify a new molecular mechanism involved in Rh-deficiency. The first mutation affected the invariant G residue of the 3' acceptor splice-site of intron 6, causing the skipping of the downstream exon and the premature termination of translation. The second mutation occurred at the first base of the 5' donor splice-site of intron 1. Both these mutations were found in homozygote state. RNase protection assays demonstrated that the Rh50 mRNA level was strongly reduced or undetectable in the 3' and 5' splice mutants, respectively. The different mutations affecting the RH50 gene are indicative of an heterogeneous mutational pattern, which further supports the hypothesis that the lack of the Rh50 protein may prevent the assembly or transport of the Rh membrane complex to the red blood cell surface. PMID- 9746796 TI - The PIG-A mutation and absence of glycosylphosphatidylinositol-linked proteins do not confer resistance to apoptosis in paroxysmal nocturnal hemoglobinuria. AB - Paroxysmal nocturnal hemoglobinuria (PNH) is a clonal stem cell disorder characterized by complement-mediated hemolysis and deficient hematopoiesis. The development of PNH involves an acquired mutation in the X-linked PIG-A gene, which leads to incomplete bioassembly of glycosylphosphatidylinositol (GPI) anchors and absent or reduced surface expression of GPI-linked proteins. The origin and mechanisms by which the PNH clone becomes dominant are not well understood, but recently resistance to apoptosis has been postulated. To test the hypothesis that the PIG-A mutation and absence of GPI-linked surface proteins directly confer resistance to apoptosis, we isolated peripheral granulocytes from 26 patients with PNH and 20 normal controls and measured apoptosis induced by serum starvation. Granulocytes from patients with PNH were relatively resistant to apoptosis (38.8% +/- 14.1%) as compared with granulocytes from controls (55.0% +/- 12.0%, P < .001). However, this resistance to apoptosis was not related to the dominance of the PNH clone because patients with a low percentage of GPI deficient granulocytes had a similar rate of apoptosis as those with a high percentage of GPI-deficient granulocytes. Similarly, the resistance to granulocyte apoptosis was not influenced by the degree of neutropenia or a prior history of aplastic anemia. To investigate formally the importance of GPI-linked surface proteins in apoptosis, we introduced the PIG-A cDNA sequence into the JY5 GPI-negative B-lymphoblastoid cell line using two different methods: (1) stable transfection of a plasmid containing PIG-A, and (2) stable transduction of a retroviral vector containing PIG-A. We then measured rates of apoptosis induced either by Fas antibody, serum starvation, or gamma-irradiation. With each stimulus, apoptosis of JY5 with stable surface expression of GPI-linked proteins was not statistically different from the parent JY5 cell line or the JY25 (GPI positive) cell line. Our data confirm that granulocytes from patients with PNH have a relative resistance to apoptosis as compared with normal granulocytes. However, this resistance does not vary with the level of expression of GPI-linked proteins, and stable introduction of PIG-A cDNA with correction of GPI-linked surface expression does not change the rate of apoptosis. Taken together, our data do not support the hypothesis that the PIG-A mutation and absence of GPI linked surface proteins directly confer resistance to apoptosis in PNH. We conclude that the resistance to apoptosis in PNH is not related to the PIG-A mutation, indicating that other factors must be important in the origin of this phenomenon and the clonal dominance observed in PNH. PMID- 9746797 TI - Plasma endothelin-1, cytokine, and prostaglandin E2 levels in sickle cell disease and acute vaso-occlusive sickle crisis. AB - The relative contributions of microvascular inflammation and vasomotor dysregulation to the development of acute vaso-occlusive crisis in sickle cell disease have been intensely studied. The present observational study was designed to examine the levels of circulating proinflammatory cytokines, anti-inflammatory cytokines, and vasoactive mediators during and after acute painful crisis. In symptomatic sickle cell patients, plasma levels of endothelin-1 and prostaglandin E2 were elevated during crises compared with healthy African-American controls. These levels had decreased, but not normalized, when patients were seen 1 to 3 weeks after discharge from hospital. Other mediators (tumor necrosis factor alpha [TNFalpha], interleukin-1beta [IL-1beta], IL-6, IL-8, and IL-10) were neither elevated in asymptomatic sickle cell disease nor in acute vaso-occlusive crisis. As a potent long-acting mediator of vasoconstriction and inflammation, endothelin 1 may play a key role in the cycle of ischemia and inflammation that initiates and sustains pain of crisis. The downregulatory effects of prostaglandin E2 on immune cell function may contribute to the increased susceptibility to infection observed in patients with sickle cell disease. PMID- 9746798 TI - Nonobese diabetic/severe combined immunodeficiency (NOD/SCID) mouse as a model system to study the engraftment and mobilization of human peripheral blood stem cells. AB - Mobilized CD34(+) cells from human peripheral blood (PB) are increasingly used for hematopoietic stem-cell transplantation. However, the mechanisms involved in the mobilization of human hematopoietic stem and progenitor cells are largely unknown. To study the mobilization of human progenitor cells in an experimental animal model in response to different treatment regimens, we injected intravenously a total of 92 immunodeficient nonobese diabetic/severe combined immunodeficiency (NOD/SCID) mice with various numbers of granulocyte colony stimulating factor (G-CSF) -mobilized CD34(+) PB cells (ranging from 2 to 50 x 10(6) cells per animal). Engraftment of human cells was detectable for up to 6.5 months after transplantation and, depending on the number of cells injected, reached as high as 96% in the bone marrow (BM), displaying an organ-specific maturation pattern of T- and B-lymphoid and myeloid cells. Among the different mobilization regimens tested, human clonogenic cells could be mobilized from the BM into the PB (P = .019) with a high or low dose of human G-CSF, alone or in combination with human stem-cell factor (SCF), with an average increase of 4.6 fold over control. Therefore, xenotransplantation of human cells in NOD/SCID mice will provide a basis to further study the mechanisms of mobilization and the biology of the mobilized primitive human hematopoietic cell. PMID- 9746799 TI - Host reactive donor T cells are associated with lung injury after experimental allogeneic bone marrow transplantation. AB - Noninfectious lung injury is common after allogeneic bone marrow transplantation (BMT), but its association with acute graft-versus-host disease (GVHD) is unclear. Using a murine BMT system where donor and host differ by multiple minor histocompatibility (H) antigens, we investigated the nature of lung injury and its relationship both to systemic GVHD and host-reactive donor T cells. Lethally irradiated CBA hosts received syngeneic BMT or allogeneic (B10.BR) T-cell depleted (TCD) bone marrow (BM) with and without the addition of T cells. Six weeks after BMT, significant pulmonary histopathology was observed in animals receiving allogeneic BMT compared with syngeneic controls. Lung damage was greater in mice that received allogeneic T cells and developed GVHD, but it was also detectable after TCD BMT when signs of clinical and histologic acute GVHD were absent. In each setting, lung injury was associated with significant alterations in pulmonary function. Mature, donor (Vbeta6(+) and Vbeta3(+)) T cells were significantly increased in the broncho-alveolar lavage (BAL) fluid of all allogeneic BMT recipients compared with syngeneic controls, and these cells proliferated and produced interferon-gamma (IFN-gamma) to host antigens in vitro. These in vitro responses correlated with increased IFN-gamma and tumor necrosis factor-alpha (TNF-alpha) in the BAL fluid. We conclude that alloreactive donor lymphocytes are associated with lung injury in this allogeneic BMT model. The expansion of these cells in the BAL fluid and their ability to respond to host antigens even when systemic tolerance has been established (ie, the absence of clinical GVHD) suggest that the lung may serve as a sanctuary site for these host reactive donor T cells. These findings may have important implications with regard to the evaluation and treatment of pulmonary dysfunction after allogeneic BMT even when clinical GVHD is absent. PMID- 9746800 TI - A role for transforming growth factor-beta1 in the increased pneumonitis in murine allogeneic bone marrow transplant recipients with graft-versus-host disease after pulmonary herpes simplex virus type 1 infection. AB - To gain further insights in the pathogenesis of herpesvirus pneumonia in allogeneic bone marrow transplant recipients, transplanted mice (B10.BR --> CBA) with graft-versus-host disease (GVHD) and control mice (transplanted mice without GVHD and normal CBA mice) were infected intranasally with herpes simplex virus type 1 (HSV-1). When compared with infected control mice, infected allogeneic transplant recipients with GVHD showed increased periluminal mononuclear cell infiltrates. However, infected allogeneic transplant recipients with GVHD showed lower virus content in the lung tissue than infected control mice. High concentrations of transforming growth factor-beta 1 (TGF-beta1) were detected in the bronchoalveolar lavage (BAL) fluid of mock-infected allogeneic transplant recipients with GVHD, which increased slightly after infection. Anti-TGF-beta treatment of allogeneic transplant recipients with GVHD significantly decreased the histological evidence of pneumonitis at day 4 after HSV-1 infection. We conclude that allogeneic transplant recipients with GVHD have (1) increased pneumonia, (2) highly elevated levels of TGF-beta1 in the BAL fluid, and (3) reduced pulmonary virus content after HSV-1 infection. Our data suggest that the newly recognized dysregulation of cytokine (TGF-beta1) production may be more important than the viral load for the increased severity of HSV-1 pneumonia in allogeneic transplant recipients with GVHD. PMID- 9746802 TI - Reduced spectrin-ankyrin binding in a South African hereditary elliptocytosis kindred homozygous for spectrin St Claude. PMID- 9746801 TI - How old are dense red blood Cells? The dog's tale. PMID- 9746803 TI - Genetic heterogeneity of congenital dyserythropoietic anemia type II. PMID- 9746804 TI - Increased levels of endothelin-1 in plasma of sickle cell anemia patients. PMID- 9746805 TI - Activating mutations of the transmembrane domain of MPL in vitro and in vivo: incorrect sequence of MPL-K, an alternative spliced form of MPL. PMID- 9746806 TI - Prediction of human herpesvirus 6 infection after allogeneic bone marrow transplantation. PMID- 9746808 TI - Evidence for genetic homogeneity in a familial platelet disorder with predisposition to acute myelogenous leukemia (FPD/AML) PMID- 9746807 TI - Clinical significance of Arg306 mutations of factor V gene. PMID- 9746809 TI - Human RhDel is caused by a deletion of 1,013 bp between introns 8 and 9 including exon 9 of RHD gene. PMID- 9746810 TI - Sterilisation of married couples: husband versus wife sterilisation. AB - Sterilisation has been increasing in the United States in recent decades. Using the National Survey of Families and Households, this paper examines sterilisation among married couples using event history techniques, viewing husband and wife sterilisation as competing risks. Wives are more likely to experience sterilisation and at shorter durations of marriage. Number of children has a curvilinear effect on sterilisation, increasing and then decreasing its likelihood. Wives who are older than their husbands are more likely to get sterilised themselves. Black and Hispanic husbands are more likely to undergo sterilisation. PMID- 9746811 TI - The interacting effects of prices and weather on population cycles in a preindustrial community. AB - The exogenous cycles and population dynamics of the community at Penrith, Cumbria, England, have been studied (1557-1812) using aggregative analysis, family reconstitution and time series analysis. This community was living under marginal conditions for the first 200 years and the evidence presented is of a homeostatic regime where famine, malnutrition and epidemic disease acted to regulate the balance between resources and population size. This provides an ideal historic population for an investigation of the direct and indirect effects of malnutrition. Throughout the period studied, a short wavelength oscillation in grain prices was apparently the major external factor that drove exogenous cycles in mortality, birth rate, and migration. In particular, the different responses of children to variations in food supply are emphasised; fluctuations in poor nutrition correlated significantly with the variations in mortality rates for infants (probably indirectly during pregnancy and directly during the first year of life) and for young children (via susceptibility to lethal infectious diseases). Migratory movements contributed to the maintenance of homeostasis in the population dynamics. A medium wavelength cycle in low winter temperatures was associated with a rise in adult mortality which, in turn, promoted an influx of migrants into this saturated habitat. A model incorporating these interacting associations between vital events and exogenous cycles is presented: grain prices were an important density-dependent factor and constituted the major component of the negative feedback of this population and drove the exogenous, short wavelength mortality cycles. Cycles of births and immigration provide a positive feedback for the build-up of susceptibles and the initiation of smallpox epidemics and increased population size. PMID- 9746812 TI - Stability of marital unions and fertility in Nigeria. AB - Using nationally representative data, it is shown that marital unions are relatively stable in Nigeria. Remarriage rates are high so little time is lost between unions. Consequently, the fertility of women who have experienced marital disruption is only slightly lower than for those in stable unions. Their slightly lower parity may be a function of a high incidence of reproductive impairment, which is a major reason for divorce and separation in Nigeria. PMID- 9746813 TI - Some factors affecting marital distances in the Outer Hebrides. AB - Some factors affecting marital distances have been studied in two Outer Hebridean islands, Harris (843 marriages) and Barra (444 marriages), over the period 1855 1990. In each island marital distances fell before 1900, but then rose to their greatest values after the 1950s. Fisherman generally married at the shortest distances and men in land-based occupations at the longest. The depression in the fishing industry during the 1880s and early 1890s was associated with reductions in marital distances, especially among fisherman. In the different regions of Harris, marital distances were least in the south-east, where settlement was most dense, and greatest in the south-west where it was most sparse. When the association between marital frequencies and inter-settlement distances was studied, it was found that for Harris there was, overall, a trend to endogamous and short-distance marriage. However, this trend was only slight during 1955-90. For Barra a similar trend was found before 1955, but thereafter there appeared to be virtually no connection between marital frequency and inter-settlement distance. Thus the only constraint on marriage was the spatial distribution of settlements. In this situation the chances of random mating with respect to distance are maximised. Application of 'Central Place' theory suggested that only since 1946 can any tendency be detected to regard Tarbert in Harris or Castlebay in Barra as Central Places, at least as far as marriage is concerned. In each island the tendency appears to be limited to the settlements closest to the Central Place. PMID- 9746814 TI - Sexual networks and the transmission of HIV in London. AB - This paper discusses ways in which empirical research investigating sexual networks can further understanding of the transmission of HIV in London, using information from a 24-month period of participant observation and 53 open-ended, in-depth interviews with eighteen men and one woman who have direct and indirect sexual links with each other. These interviews enabled the identification of a wider sexual network between 154 participants and contacts during the year August 1994-July 1995. The linked network data help to identify pathways of transmission between individuals who are HIV+ and those who are HIV-, as well as sexual links between 'older' and 'younger' men, and with male prostitutes. There appears to be considerable on-going transmission of HIV in London. The majority of participants reported having had unprotected anal and/or vaginal sex within a variety of relationships. The implications of these findings for policies designed to prevent the transmission of HIV are discussed. PMID- 9746815 TI - An ecologic study of the relationship between mean birth weight, temperature and calorie consumption level in Japan. AB - This study reports an ecologic analysis of the relationship between mean birth weight (MBW) and nutritional, medical, and social variables, using 1982 data for 47 prefectures in Japan. Correlation analysis showed that variables that correlated significantly with MBW were mean temperature (r = 0.63), total calorie intake (r = 0.56), and body mass index of women (r = 0.60). It is concluded that the MBW level in a given community represents the long and short term nutritional condition of mothers, and that the level may follow Bergman's law: 'In the same species, the body size of an animal increases along with latitude; that is, the lower the temperature, the larger the body size.' PMID- 9746816 TI - Sex on campus: a preliminary study of knowledge, attitudes and behaviour of university students in Delhi, India. AB - Eight hundred and eighty-seven students from two major universities in Delhi, India, were surveyed, using a self-administered questionnaire, about their sexual knowledge, attitudes and behaviour. The data show that female students seem to be rejecting traditional Indian repressive sexual standards of premarital and non procreative sex and the gender differences are beginning to narrow. Despite their sexual awareness, the students were highly ignorant of the facts of life. Being male and married did not make them more knowledgeable. PMID- 9746817 TI - Infant feeding practices and child health in Bolivia. AB - The effects of breast-feeding and supplementation practices on recent diarrhoea occurrence and stunted growth are modelled using logistic regression techniques. Data from the Demographic and Health Survey of Bolivia, 1989, show that, among children aged 3-36 months at the date of interview, the benefits of breast feeding to child health were most pronounced among children living in rural poverty. Reduced breast-feeding among these children increased the likelihood of diarrhoea and stunted growth. In addition, the introduction of solid foods to currently lactating infants negatively influenced child health. PMID- 9746818 TI - Infertility treatment and multiple birth rates in Britain 1938-94. A comment. PMID- 9746819 TI - Coital frequency among married and cohabiting couples in the United States. A comment. PMID- 9746820 TI - Prior and proximate causes of infant survival in Ghana, with special attention to polygyny. A comment. PMID- 9746821 TI - The impact of nutritional status on body fat distribution patterns in pre- and postmenopausal females. AB - This study examines the impact of nutritional status, classified by body mass index, on sex specific fat distribution patterns dependent on menopausal status in 467 pre-, peri- or postmenopausal females. Absolute and relative amounts of upper and lower body fat were estimated by means of dual energy X-ray absorptiometry. It was found that low weight, independent of menopausal status, leads to the typical gynoid pattern of fat distribution while excess weight and obesity result in the android pattern of distribution in pre- and postmenopausal women. PMID- 9746822 TI - Impact of migration, environment and socioeconomic conditions on blood pressure of Sikhs. AB - This study documents the impact of migration, environment and caste on the blood pressure of Sikhs living in the UK and their peers in the native Punjab state of India. A sample of 449 adult Sikhs, males and females, were studied in the Punjab state, and in Southall, in the Ealing borough of London and Handsworth in Birmingham in the UK. There is minor variation in the blood pressure of members of various castes among the Sikhs living in the Punjab. The pattern in blood pressure broadly corresponds with the economic status of the individuals, with well-off groups showing higher blood pressure compared to the poorer groups. The blood pressure of migrants, though higher than that of their sedente peers, is not significantly so. Age at arrival in the UK and the length of exposure to the new environment were generally not significantly related to variation in blood pressure. There seems to be a secular shift in the blood pressure values of the native Punjabi Sikhs compared to earlier studies, possibly because of the rise in civic disturbances and social unrest in the Punjab during recent years. PMID- 9746823 TI - Motives underlying healthy eating: using the Food Choice Questionnaire to explain variation in dietary intake. AB - The Food Choice Questionnaire (FCQ), which measures the reported importance to a given individual of nine factors underlying food choice, and a food frequency questionnaire, were administered to 241 participants, who were also required to classify their diet as either 'standard', 'low in red meat' or 'vegetarian'. Respondents describing their diet as low in red meat attributed greater importance to health, natural content, weight control and ethical concern in their food choice than did those who described their diets as standard, whereas vegetarians differed significantly from those with a standard diet only on the score for ethical concern. Differences between men and women and between students and non-students in the frequency of consumption of a number of foods were shown to be mediated by differences in the importance attached to FCQ factors. Thus the generally healthier diets of women compared to men appeared to be accounted for by the greater importance attributed by women to weight control, natural content and ethical concerns. PMID- 9746824 TI - Assessing and interpreting birth spacing goals in Costa Rica. AB - A procedure for assessing birth spacing goals, an important component of fertility preferences, is proposed and applied to 1993 Costa Rican data. Based on a reverse or backward survival analysis, preferred birth intervals are estimated to range between 3.5 and 4.5 years (1.5 years for the interval union to first birth). These intervals are 2 or 3 years shorter than crude estimates from data on open or last closed intervals, which are upwardly biased by selection and left censoring effects. To achieve these spacing preferences, a cohort must spend about two-thirds of the time using contraception (one-third in the interval union to first birth). An inverse association between desired family size and desired birth interval is evident only in parity-specific analyses. PMID- 9746825 TI - From efficacy to effectiveness: selecting indicators for a community-based lactational amenorrhoea method promotion programme. AB - This paper reviews the results of clinical trials and community studies of lactational amenorrhoea and its role as a contraceptive method (LAM). Indicators which are used in efficacy trials and effectiveness interventions are compared and sets of indicators of effectiveness appropriate to community-based LAM programmes are recommended. A five-tiered ecological framework is used to facilitate selection of indicators which range from individual to policy level outcomes. The indicator framework is intended as a tool for health practitioners in family planning and maternal and child health service delivery settings who are interested in designing programmatic interventions for the promotion of LAM, particularly among less well-educated women of lower socioeconomic communities. PMID- 9746826 TI - Postpartum amenorrhoea in rural eastern Uttar Pradesh, India. AB - This paper calculates the mean duration of the postpartum amenorrhoea (PPA) and examines its demographic, and socioeconomic correlates in rural north India, using data collected through 'retrospective' (last but one child) as well as 'current status' (last child) reporting of the duration of PPA. The mean duration of PPA was higher in the current status than in the retrospective data; the difference being statistically significant. However, for the same mothers who gave PPA information in both the data sets, the difference in mean duration of PPA was not statistically significant. The correlates were identical in both the data sets. The current status data were more complete in terms of the coverage, and perhaps less distorted by reporting errors caused by recall lapse. A positive relationship of the mean duration of PPA was found with longer breast-feeding, higher parity and age of mother at the birth of the child, and the survival status of the child. An inverse relationship was found with higher education of a woman, higher education of her husband and higher socioeconomic status of her household, these variables possibly acting as proxies for women's better nutritional status. PMID- 9746827 TI - Water contact patterns and behavioural knowledge of schistosomiasis in south-west Nigeria. AB - Human water contact patterns were studied in two resettlement communities at the Oyan Reservoir in south-west Nigeria in all four seasons in 1991 and 1992. Water contact was most intensive in the afternoon and in the hot dry season, but different types of activities exhibited different daily and seasonal patterns. Both communities were highly endemic for urinary schistosomiasis. However, knowledge regarding schistosomiasis transmission was very limited and the infection was, in spite of a very high frequency of blood in the urine, not considered a major public health problem. Most water contacts were of either a recreational (swimming, bathing) or economic (fishing) nature, and age- and sex related patterns were evident. The overall level of exposure peaked in the 10-14 years age group but water contact of an economic nature peaked in the 20-39 years age group. Females had generally more water contact than males. PMID- 9746828 TI - The prevalence and demographic characteristics of consanguineous marriages in Pakistan. AB - Consanguineous marriages are strongly preferred in much of West and South Asia. This paper examines the prevalence and sociodemographic correlates of consanguineous unions in Pakistan using local and national data. Information from 1011 ever-married women living in four multi-ethnic and multi-lingual squatter settlements of Karachi, the main commercial centre of the country, are compared with data from the national 1990/91 Pakistan Demographic and Health Survey (PDHS), based on information provided by 6611 women. Both sets of results indicate that approximately 60% of marriages were consanguineous, over 80% of which were between first cousins. The mean coefficients of inbreeding (F) in the present generation were 0.0316 and 0.0331 for the Karachi and PDHS data respectively. In both surveys the prevalence of consanguineous unions appeared to be unchanged over the past three to four decades. Consanguineous unions were more common among women who were illiterate or had only primary level education, were first or second generation migrants from rural areas of Pakistan or, in the PDHS, lived in rural areas, and whose parents were also consanguineously married. PMID- 9746829 TI - Contraceptive use and desire for more children in two rural districts of Sierra Leone. AB - Data from a 1993 household survey in rural Sierra Leone revealed that, among women aged 12-49 with at least one child younger than 5, about 13% were using a contraceptive method and about 67% wanted more children. These rates differ from those reported for the 1980s and 1970s, when the contraceptive use rate was around 6% and more than 85% of women desired more children, suggesting a trend towards fertility limitation over the years. Logistic regression analysis showed that contraceptive use was positively associated with age, number of living children, age at marriage, education, and economic status, and negatively associated with Islamic and traditional religious affiliations. Government and other health clinics, hospital, and government paramedical personnel were the major sources of contraceptive supplies. The lower desire for more children might relate to demographic pressure from the recent improved child survival rate compounded by recent economic hardship. PMID- 9746830 TI - Factors influencing continuation of IUD use in south India: evidence from a multivariate analysis. AB - This paper studies the correlates of IUD continuation, particularly in relation to quality of service provided in Karnataka, South India, by using a discrete time logit model. Provision of follow-up services had a moderate influence on continuation, and medical check-up at insertion influenced the experience of side effects. But these effects were trivial in comparison with the overriding influence of motivational variables and of reported side effects. The IUD is retained for a limited duration in rural India where it is used mainly as a spacing device by lowly motivated young women who discontinue the method at the slightest feeling of discomfort or abnormality. If the IUD were recommended to older women for limiting childbearing by emphasizing its reversibility, it would enhance the method's popularity and improve the levels of contraceptive use among younger women. PMID- 9746831 TI - Seasonality of induced abortion in North Carolina. AB - This paper examines the seasonality of induced abortion in North Carolina between 1980 and 1993. Distinct seasonal patterns are found, with a peak in February and a valley in September. These patterns correspond to the implicit seasonality of conceptions associated with the seasonality of birth pattern. One notable difference from the general pattern is among unmarried women aged 18 and younger. They have the February peak and an additional peak in August that may be associated with the summer vacation from school. PMID- 9746832 TI - Infant and child mortality in Bangladesh: age-specific effects of previous child's death. AB - This study examines whether mortality of two adjacent siblings in families is age specific and is modified by the MCH-FP programme and fertility and mortality declines in Matlab, Bangladesh, using data for singleton births during 1977-78, 1985-86 and 1989-90 in the treatment (MCH-FP) and comparison areas. Logistic regression was used to estimate the net effects of survival status of elder siblings on mortality of younger siblings in the neonatal, postneonatal and toddler periods, controlling for birth order, previous birth interval, maternal age, education and religion, household possession of valuable items and sex of the child. Odds of neonatal and postneonatal deaths of younger siblings were found to be higher if the elder sibling had died at the same age than if the sibling had survived infancy. Toddler mortality was lower if the elder sibling had died in infancy. The association between two siblings'mortality risks did not decline over time in either area. The results suggest that a family history of child deaths by age is important to identify when subsequent infants would be at a higher risk of dying. PMID- 9746833 TI - Chronic energy deficiency in women from rural Bangladesh: some socioeconomic determinants. AB - This paper explores a number of socioeconomic factors thought to explain the wide prevalence of undernutrition among rural Bangladeshi women. The 1992 baseline survey data of the BRAC-ICDDR,B Joint Research Project at Matlab were used. Anthropometry was performed on a random sub-sample of 1462 currently married, non pregnant women between 15 and 49 years of age. Women's nutritional status was defined in terms of Body Mass Index (BMI = wt in kg/ht in m2). Compared with women from better-off households, the mean weight (41.2 vs 43.0 kg; p < 0.0001), mid-upper arm circumference (MUAC) (22.1 vs 22.7; p < 0.0001), and BMI (18.5 vs 19.1; p < 0.0001) of poor women were consistently lower. However, no significant difference in mean height was found between the two groups. The results showed that women aged more than 35 years are twice as likely to have a BMI < 18.5 compared with younger women. Both years of schooling received and socioeconomic status are found to be important predictors of women's BMI. Women who have received one or more years of formal education are nearly half as likely to suffer chronic energy deficiency (BMI < 18.5) than women with no schooling. Again, better-off women are found to be 0.77 times less likely to have chronic energy deficiency than women from poor households. The implications of these findings in improving the nutritional status of rural Bangladeshi women are discussed. PMID- 9746834 TI - Genetic drift in present human populations: survey of a Mediterranean region (La Alpujarra, South-east Spain). AB - This paper presents information on the opportunity for genetic drift in La Alpujarra, a rural population of the Spanish Mediterranean. This region is characterized by its historical isolation. The analyses were based on a survey of 45% of the population. The Effective Population Size (NE = 1986), the Effective Migration Rate (M(E) = 12.4) and the Coefficient of Breeding Isolation (CBI = 246.2) of Lasker & Kaplan (1964) were calculated. The value of the latter index indicates that genetic drift in the Alpujarrenian population is negligible at present because of the cessation of reproductive isolation due to the development of communication networks. PMID- 9746835 TI - A demographic and health survey of Spanish female sex workers: HIV prevalence and associated risk factors. AB - In order to examine the prevalence of HIV infection and associated risk factors amongst Spanish female prostitutes a multicentre (n = 22) cross-sectional study was conducted between November 1989 and January 1991. Data collected included socioeconomic characteristics, sexual behavior and health status together with blood samples. A total of 1633 prostitutes were recruited into the study. Of these, 1433 (87.8%) consented to blood samples being taken and 180 (12.6%) were HIV positive. HIV seroprevalence was 54.7% for intravenous drug users (IVDUs) versus 3.7% for non-IVDUs. Previous imprisonment, hepatitis B and a partner who was an IVDU were significant predictors of HIV infection according to multivariate logistic regression models. PMID- 9746836 TI - Attitudes of men towards family planning in Mbeya region, Tanzania: a rural-urban comparison of qualitative data. AB - Family planning programmes in Tanzania date back to the 1950s. By the early 1990s, however, only 5-10% of women of childbearing age used contraceptives in the country. Low contraceptive prevalence in Tanzania is reportedly attributable to men's opposition to family planning. This paper employs focus groups to explore the role of Tanzanian men in family planning. More specifically, it presents a rural-urban comparison of the attitudes of men in Mbeya region, Tanzania, to family size preference, sex composition, partners' communication on family planning matters and contraceptive behavior. Findings indicate that men express positive attitudes towards fertility-regulating methods. There is, moreover, little rural-urban variation in male attitudes towards family planning in the study area. Possible reasons for this normative convergence (including structural similarities and rural-urban migration between the two communities) are discussed. PMID- 9746837 TI - Reproduction, risk and reality: family planning and reproductive health in northern Vietnam. AB - In collaboration with the National Committee for Population and Family Planning, a study was conducted in 1994 in two Vietnamese communes to provide community level information on women's reproductive health and behaviors. A survey of 504 rural and 523 urban women collected five-year histories of reproduction, contraception, abortion and symptoms of Reproductive Tract Infections (RTI). This analysis focuses on the relationships between women's individual characteristics, use of family planning and abortion, and reported RTI symptoms. The findings reveal that IUDs do not raise women's likelihood of experiencing RTI symptoms in either commune. A recent abortion, however, strongly increases women's likelihood of having RTI symptoms in the rural commune, while low-socioeconomic status is associated with RTI symptoms in the urban commune. PMID- 9746838 TI - Impact of rural-urban migration on fertility: a population ecology analysis in the Kombio, Papua New Guinea. AB - The Anjangmui dialect group of the Kombio in Papua New Guinea has experienced a rapid increase in rural-urban migration since European contact commenced in the 1930s. Population ecology analyses of birth and migration histories for 240 Anjangmui women showed a higher total marital fertility rate in the cohort born in 1940-59 than that born in 1920-39. A decline in the age at menarche for nutritional reasons, and reduction in the birth interval for behavioral reasons, may explain the fertility difference between cohorts. Comparison of age-specific marital fertility rates between migrants in urban areas and non-migrants in rural areas revealed higher rates among migrants in the 15-19 and 20-24 year age groups, but lower rates in the 25-29 year or older age groups; the total marital fertility rate for migrants was lower than that for non-migrants. The differences may be attributable to the different influences of birth control practices on fertility reduction between the migrants in urban areas and non-migrants in rural areas. It is suggested that rural-urban migration in the Anjangmui during the post-contact period has had the effect of reducing fertility in the population as a whole. PMID- 9746839 TI - Risk factors of low birthweight. A comment. PMID- 9746840 TI - [Therapy for pain: oxycodon--a new delayed-action opioid with effective analgesic action]. PMID- 9746841 TI - [How dangerous are high-resistance bacteria in intensive care units?]. PMID- 9746842 TI - [NMDA-receptor antagonist in pain therapy]. AB - The treatment of patients suffering from chronic neuropathic pain remains a clinical challenge, particularly in cases where opioid therapy fails to provide sufficient pain relief. Spinal sensitization might be one cause for induction and maintenance of such states of pain, frequently accompainied by symptoms like allodynia, hyperalgesia and temporal summation of second pain. Experimental data concerning the role of NMDA-mediated processes in central sensitization and the effects of NMDA receptor antagonists in different models of neuropathic pain are reviewed. In clinical trials ketamine and other NMDA receptor blocking agents caused a significant reduction of hypersensitive states of pain, but nearly all authors described psychomimetic and other side effects. Aminoadamantanes like memantine and amantadine also have NMDA blocking properties and are widely used in the treatment of Parkinson's disease. Further clinical studies may reveal whether these substances will play a role as adjuncts in future pain treatment. Improving the efficacy of opioids by blocking NMDA receptor-mediated activity constitutes another clinically relevant concept for pain management. Numerous experiments have shown synergistic effects of NMDA antagonists and opioids in analgesia, while the development of opioid tolerance was prevented. PMID- 9746843 TI - [Use of the laryngeal mask in oral and dental surgery]. AB - OBJECTIVE: The suitability of the laryngeal mask (LM) for anesthesia in oral and dental surgery of the face was investigated considering in particular the risks of aspiration and dislocation. We also examined acceptability to the surgeon. METHODS: In a prospective study, a total of 105 patients (ASA I-III) was included. Provided with flexible LMs, patients were operated upon the third molars (n = 64), around the dentoalveolar area (n = 32) and extraorally (n = 9). The number of placing efforts, preoperative leakage pressure, cuff pressure and complications occurring during the course of anesthesia were recorded. After the operation, the procedure was compared to endotracheal anesthesia (ETA) by the surgeon. A retrospective study comparing LM with ETA was performed on 1395 patients examining the time in between two operations, the period from the end of operation to the beginning of control of vital signs in the recovery room, and the time spent in the recovery room. The number of patients necessitating a change of anesthesia from LM to ETA was recorded. RESULTS: In 33 patients (31%), intraoperative leakage due to dislocation was observed. 33 patients (31%) had obstruction of the airway. Only when leakage occurred together with obstruction of the airway, SaO2 fell below 94% (n = 11). This was seen in particular during exposure of the wisdom teeth. In 2 of these cases, SaO2 decreased below 70 or 34% respectively. Aspiration of blood, gastric juice or dental and bone fragments was not observed. In one case, ETA became necessary. 19 patients complained of local pains (cough, sore throat, dysphagia). Operative conditions for the surgeon were comparable to oral ETA in 79% of the cases. The time in between two operations decreased about 35%, the period from the end of operation until first control of vital signs in the recovery room was reduced of about 41%, and the time spent in the recovery room decreased by 37% when compared to ETA. Out of 1111 anesthesias performed with the LM, 17 had to be exchanged for ETA. CONCLUSION: Leakage due to dislocation of the LM and airway obstruction only leads to a relevant risk of SaO2 to drop if both components occur simultaneously. With increasing experience and close cooperation between surgeon and anesthesiologist, they may be almost completely avoided so that even exposure of the lower, wisdorri teeth will seldom iiecessiiate the LM to be replaced by ETA. In relation to the tongue depressor, cuff pressure should be kept at low levels in order to obtain better flexibility of the LM. The LM provides sufficient protection against aspiration of intraoperative fluid in the pharynx. Acceptability to the surgeon is high because of good operative conditions and shortened periods in between two operations when compared to ETA. Improved protective reflex responses after the operation and its advantages when used in patients with tightness of the jaw make the LM a suitable instrument for anesthesia in oral and dental surgery. PMID- 9746845 TI - [Methicillin-reistant Staphylococcus aureus: risk factors for infection colonization and clonal heterogeneity in intensive care units]. AB - PURPOSE: The aim of this study was to determine the risk factors associated with colonisation/infection by methicillin-resistant Staphylococcus aureus (MRSA) and to demonstrate the chain of infections by genotyping of all MRSA isolates. METHODS: A total of 6143 microbiological samples from 1753 patients was obtained at a surgical ICU of Frankfurt University hospital during 1995. RESULTS: MRSA was detected in 1.6% of patients and three members of staff (3.3%). Typing of these 31 MRSA-strains (first isolates) by macrorestriction analysis of chromosomal DNA revealed nine different genotypes. More than 60% of all isolates belonged to one type that was confirmed to be closely related to the "South Germany" epidemic strain. A strong correlation between severity of underlying disease and length of hospitalisation on the one hand and detection of MRSA on the other could be demonstrated. Detection of MRSA was significantly more common in patients with adult respiratory distress syndrome (ARDS), sepsis, kidney failure, prolong respiratory treatment and in patients with prolonged phases of haemodynamic instability. It appeared that the mortality rate of MRSA-infected patients was higher (28.6%) than the mortality rate of all patients (6.5%). CONCLUSION: Following of a strict hygiene regime is important to prevent clonal spread of MRSA and especially to protect immunocompromised patients from complicating infections. PMID- 9746844 TI - [Anesthesia in cesarean section in the FRG in 1997. Results of a nationwide survey]. AB - A survey of all German hospitals providing obstetric anaesthesia in 1997 (n = 1061, recovery rate 82% comprising 115,000 Caesarean sections revealed that most Caesarean sections (CS) are performed under general anaesthesia (GA). For elective CS, the average was 63%, and 82% for urgent (non-emergency) section. Succinylcholine is the standard neuromuscular blocker for intubation. Of the regional techniques, epidural continuous anaesthesia (ED) is preferred for elective CS (59%) over subarachnoid (SA, 10%) and combined epidural and subarachnoid anaesthesia (CSE). In urgent CS, SA is used more often (56%) than ED (42%) and CSE. With increasing number of births per year, the use of regional techniques is more common. PMID- 9746846 TI - [Alternative airways]. PMID- 9746847 TI - [New artificial respiration concepts in acute respiratory insufficiency--which will last ?]. PMID- 9746848 TI - [New artificial respiration modes]. PMID- 9746849 TI - [Liquid ventilation and surfactant]. PMID- 9746850 TI - [NO-inhalation: what are the indications?]. PMID- 9746851 TI - [Delayed lumbar epidural hematoma. Discussion of the risk factors: hypertension, anticoagulation and spinal anesthesia]. AB - This case report deals with the very rare complication of an intraspinal haematoma: a 70 year old male underwent anticoagulation because of supraventricular dysrhythmias followed by two transient ischaemic attacks. He accidentally received an intramuscular injection for tetanus prophylaxis and developed a deep intramuscular haematoma, which was operated upon, after normalisation of coagulation parameters, under spinal anaesthesia. No primary complication was noted. Heparin therapy was started perioperatively, on the day of the operation. Sixteen days postoperatively, the patient resumed oral anticoagulation; 3 months later he developed a progressive cauda-equina-syndrome due to an epidural haematoma at the level of L2 to L4. This could be diagnosed by magnetic resonance imaging, but not by computed tomography. Acute surgical decompression was performed. The haematoma showed various ages as suspected by the intraoperative morphology and proven by histological examination. The neurological findings improved, and 6 months after rehabilitation only slight neurological deficits remained. Hypertension, anticoagulants, and spinal anaesthesia are discussed as risk factors for this complication. PMID- 9746852 TI - [25th Neonatal and Infant Respiration Symposium. March 10-14, Vail, Colorado]. PMID- 9746853 TI - [Fulminant gas embolism in arthroscopy of the shoulder joint from use of CO2. Ch. Bauereis, A. Schifferdecker, J. Buttner, H. Hempfling: AINS 199631: 654-657]. PMID- 9746855 TI - Autoantibodies in systemic lupus erythematosus. AB - Cationic germ line gene-encoded anti-DNA antibodies appear to cause inflammatory lesions via deposition and in situ formation of immune complexes, but also perhaps through apoptosis of glomerular mesangial cells. Somatic mutation is not critical for the expression of the electrical net charge of anti-DNA. In one transgenic mouse model, the nephrogenicity of anti-DNA was dependent on the expression of interleukin alpha by epidermal cells. Anti-P autoantibodies are present in the serum of healthy children but are masked by IgG antibodies. Some anti-P antibodies appear to be a subset of antilymphocyte autoantibodies. It was confirmed that anti-Ro/SS-A antibodies react with structurally unrelated conformational epitopes. The combined results of immunofluorescence and enzyme linked immunosorbent assay in the detection of antineutrophil cytoplasmic antibody has a 99.5% specificity for vasculitis among patients with connective tissue diseases. Antinucleosome antibodies are highly prevalent in patients with systemic lupus erythematosus and show an inverse correlation with nucleosome plasma levels. Transfected T-cell receptor alpha chains specific for nucleosomal autoepitopes from a Th clone that accelerates lupus nephritis, recognized the nucleosomal epitopes presented by antigen presenting cell-bearing 1-A molecules, as well as human DR molecules, via the classical major histocompatibility complex class II groove. IgG antibodies against (beta 2 -glycoprotein (GP)-1 are significantly associated with thrombosis in patients with antiphospholipid syndrome, with a specificity and sensitivity that ranges from 85% to 98% and 32% to 54%, respectively. The lupus anticoagulant activity, or the lack of it, of anti-beta 2-glycoprotein-1 appears to reside on the epitope specificity. Beta 2 glycoprotein-1 binds to apoptotic cells; in turn, anti-beta 2-glycoprotein-1 facilitate apoptotic cell clearance by macrophages. The recently described anti A2/RA33 autoantibody appears to recognize an epitope capable of distinguishing patients with lupus or with rheumatoid arthritis from those with mixed connective tissue disease. PMID- 9746856 TI - Lymphocytes, cytokines, inflammation, and immune trafficking. AB - During the past year several novel reports have added new knowledge to our understanding of the pathogenesis of system lupus erythematosus (a) a novel pathway for the presentation of autoantigens to autoreactive T cells was revealed. Universally binding nucleosomal epitopes are productively recognized by autoreactive T cells by binding to the T-cell receptor-alpha chain; (b) circulating T cells from patients with lupus commonly display a deficiency of the T-cell receptor zeta chain, and upon ligation of their cell-surface antigen receptor overproduce tyrosine phosphorylated proteins; (c) lupus and lupus nephritis are associated with a low-binding FcgammaRIIIA (CD16) polymorphism that crosses ethnic barriers; (d) the pathogenetic role of the cytokine interleukin-10 is expanding, because it is reportedly overproduced not only by cells from lupus patients but also by cells from their healthy relatives, and its overproduction in vitro is correlated with increased apoptotic cell death and with lymphopenia. PMID- 9746854 TI - The genetics of lupus. AB - Genetic factors strongly influence the risk for systemic lupus erythematosus (SLE). Studies in both animal models and humans suggest that SLE is a complex trait with contributions from multiple genes. Recent genetic studies have shown that polymorphisms at several loci, including the major histocompatibility complex, complement proteins, immunoglobulin receptors, cytokines, and other as yet unmapped genes, are associated with SLE. Ethnic factors are also important because some of these genetic associations are only found in certain populations. Murine models of SLE have provided rational candidate loci to begin genome-wide studies in humans. Promising results are now emerging from such studies. PMID- 9746858 TI - Clinical development in the management of lupus in the neonate, child, and adolescent. AB - The recent literature in pediatric aspects of systemic lupus erythematosus has focused primarily on the spectrum and management of antiphospholipid syndrome in children; this review summarizes developments in this area. In addition, the neonatal lupus syndrome is discussed in detail, and newer approaches to the management of children and adolescents with systemic lupus erythematosus will be summarized. PMID- 9746857 TI - 1998 update on antiphospholipid antibodies. AB - The field of antiphospholipid antibodies continues to evolve, with major contributions from both clinical research and laboratory studies. Antiphospholipid antibodies remain one of the more common causes of acquired hypercoagulability, both in patients with systemic lupus erythematosus and in patients who have no known connective tissue disease. In recent years, great progress has been made in identifying the protein targets. In addition, knowledge of the clinical syndrome that we call antiphospholipid antibody syndrome has also progressed. Finally, new insights into treatment continue to be gained from clinical trails. PMID- 9746860 TI - Cognitive and physical measures in rehabilitation of patients with lupus. AB - During the past 2 years, four papers describing new research studies and three papers reviewing the literature were published. These seven papers address longstanding issues, including 1) the presence or absence of cognitive impairment in systemic lupus erythematosus (SLE), 2) the prevalence of such impairment within SLE and related conditions (eg, antiphospholipid antibody syndrome, active vs inactive SLE), 3) the specific pattern of cognitive functions impaired and the possibility of a "lupus-specific" pattern, 4) the etiopathogenesis of cognitive impairments and the potential role of confounding factors such as corticosteroid use or depression, and 5) longitudinal studies examining the course of SLE related cognitive impairment. The review period also includes two studies using both static and functional neuroimaging to identify the potential relation between neuroanatomic or neurophysiologic abnormalities and cognitive impairment in patients with SLE. No papers specific to Sjogren's syndrome were identified. PMID- 9746859 TI - Therapeutic advances in systemic lupus erythematosus. AB - In treating systemic lupus erythematosus, clinicians need to consider not only the organ involvement and the complications of therapy, but also associated conditions such as premature atherosclerosis and thrombosis. A variety of agents are now available to treat cutaneous disease, including the antimalarials and thalidomide. Controversy exists about the most appropriate immunosuppressive regimen in severe disease. Experience with using cyclosporine, mycophenolate mofetil, dehydroepiandrosterone, and intravenous immunoglobulin is increasing, but plasmapheresis has not been shown to be of benefit. Autologous bone marrow transplantation has been performed. The advent of the biological era, particularly with monoclonal antibodies, gives promise of more targeted therapy. PMID- 9746861 TI - Evolving concepts of diagnosis, pathogenesis, and therapy of Sjogren's syndrome. AB - Differences in diagnostic criteria for Sjogren's Syndrome (SS) have led to confusion in the research literature and in clinical practice. A particular challenge is the clinical diagnosis of the patients with sicca symptoms, fibromyalgia, chronic fatigue, vague cognitive defects, and a low titer antinuclear antibody. Until recently, many of these patients would have been classified as primary SS using the European criteria. A suggested revision of the European criteria will require inclusion of anti SS-A antibody or characteristic minor salivary gland biopsy, leading to greater agreement between European and San Diego criteria. Recent studies have emphasized that lacrimal and salivary gland flow involves an entire "functional" unit that includes the mucosal surface (the site of inflammation), efferent nerve signals sent to the midbrain (lacrimatory and salvatory nucleus), efferent neural signals from the brain, and acinal/ductal structures in the gland. Thus, symptoms of dryness or pain can result from interferences with any part of this functional unit. The initiating antigens in SS remain unknown, but immune reactivity against SS-A, SS-B, fodrin, alpha- amylase, and carbonic anhydrase have been demonstrated in patients with established disease. The inflammatory process in the gland releases metalloproteinases that alter the relationship of epithelial cells to their matrix, an interaction that is necessary for glandular function and survival. Therapies for SS remain inadequate. In SS patients with immune-mediated extraglandular manifestation (ie, lung, kidney, skin, nerve), the therapeutic approach is similar to systemic lupus erythematosus, although these therapies have relatively little effect on tear or saliva flow. PMID- 9746862 TI - Pathogenesis and mechanisms of inflammation in the childhood rheumatic diseases. AB - The presence of CD4+ cells in the synovium of children with juvenile rheumatoid arthritis has led to the generally accepted hypothesis that aberrant activation or regulation of acquired immunity is central to the pathogenesis of this family of diseases; however, this hypothesis remains unproven, and, indeed, a specific role for T cells in the process of chronic synovitis in rheumatoid disease has yet to be identified for either adults or children. In contrast, processes associated with innate immunity are undeniably involved in the pathophysiology of chronic synovitis in both rheumatoid arthritis and juvenile rheumatoid arthritis. The presence of neutrophils in the synovial fluid, complement activation, and immune complex accumulation in the synovial fluid and serum all indicate an active inflammatory process. It is reasonable to hypothesize, therefore, that there are important clues to the cause of juvenile rheumatoid arthritis to be gleaned from a more careful study of inflammation or innate immunity. This review will focus on the roles of complement, immune complexes, and the vascular endothelium in the pathogenesis of juvenile rheumatoid arthritis. In so doing, it will reexamine the dogma of the central role of the T cell in juvenile rheumatoid arthritis disease pathogenesis and offer new paradigms with which to understand this and other rheumatic diseases of childhood. PMID- 9746863 TI - Juvenile rheumatoid arthritis and spondyloarthropathies. AB - This paper reviews the current literature on the clinical aspects of juvenile rheumatoid arthritis (JRA) and the juvenile spondyloarthropathies. The classification of the juvenile arthritides remains controversial. A sibling pair registry established a role for genetic influences on the onset and course types of JRA. Even in the absence of steroid treatment, children with JRA demonstrated decreased bone mineral density and an impairment of linear growth. Magnetic resonance imaging was found to be helpful in detecting subtalar or sacroiliac involvement. Studies were published on the use of azathioprine, cyclosporine, and cyclopyhosphamide in the treatment of severe JRA. The lack of severe liver toxicity was shown in patients having received high total doses of methotrexate. An international agreement was reached on defining improvement in JRA. Several studies found an improved long-term outcome in patients with JRA or the juvenile spondyloarthropathies. PMID- 9746864 TI - Intra-articular corticosteroids in the treatment of juvenile rheumatoid arthritis. AB - Intra-articular injection of long-acting insoluble corticosteroids produces rapid resolution of active arthritis in nearly all injected joints. Almost all of our information on the use of intra-articular corticosteroids in children comes from observational or retrospective analyses or, by inference, from studies in adult patients with arthritis. The duration of response has been found to vary according to the subtype of arthritis, the dose of injected steroids, the accuracy of injection, the duration of disease prior to injection, and possibly the age of the patient. Although the duration of follow-up in most studies has been short, intra-articular steroid therapy seems to be remarkably free of clinically important detrimental effects. Side effects are relatively uncommon and include subcutaneous atrophy and radiologically detectable structural changes or calcification. There is transient suppression of endogenous cortisol production, which may not be clinically important. Although intra-articular steroid therapy is most effective in pauciarticular juvenile rheumatoid arthritis, there are still no solid data to indicate whether it should be used earlier in the course of the disease instead of or along with systemic anti inflammatory therapy. It has been suggested that repeated injection of the same joint decreases the likelihood of a favorable response. There are still many unanswered questions about how steroids exert their beneficial effects. Newer imaging techniques promise to provide insight into the mechanism of action and possibly to a more informed basis for the use of intra-articular steroids. PMID- 9746865 TI - Childhood systemic lupus erythematosus and neonatal lupus syndrome. AB - Systemic lupus erythematosus in children can present with a wide spectrum of disease manifestations. Significant organ system involvement appears to be more severe in children than in adults. Central nervous system disease continues to be difficult to diagnose because of the lack of sensitive and specific diagnostic tests. Renal function is the major determinant of long-term prognosis and management in children with lupus. Identification of patients who are most at risk for progression of renal disease and aggressive treatment, including corticosteroids and immunosuppressive agents, are indicated. Genetic susceptibility studies in lupus reveal multiple contributions from HLA and non HLA genes. Current concepts regarding apoptosis and DNA-protein complexes and autoreactive T-cell help for anti-DNA antibody production suggest novel directions for therapies. New understandings of the pathogenesis of neonatal lupus syndrome and congenital heart block reveals important information about prospective monitoring and management of mothers and fetuses at risk. PMID- 9746866 TI - Genetic diseases with rheumatic manifestations in children. AB - Many nonrheumatic diseases of childhood present with musculoskeletal abnormalities. A significant proportion of these disorders have a genetic basis, many involving defects in structural proteins of the connective tissue. Chief among these are collagen mutations resulting in spondyloepiphyseal dysplasias and Ehlers-Danlos syndrome, as well as fibrillin defects associated with Marfan's syndrome. A variety of other chromosomal anomalies are associated with musculoskeletal abnormalities, and may result from as yet unidentified connective tissue defects. In addition, metabolic diseases may result in findings of hyper- or hypomobility, or carpal tunnel syndrome. Helpful clinical clues to identify nonrheumatologic musculoskeletal disease, as well as recent advances in our understanding of the genetic basis of several of these disorders, are reviewed here. PMID- 9746868 TI - Systemic lupus erythematosus and Sjogren's syndrome. PMID- 9746869 TI - Pediatric and heritable disorders. PMID- 9746867 TI - Bone and joint infections in children. AB - Recent reports of joint and bone infections in children have provided several new insights into prompt and accurate diagnosis of these potentially destructive processes. In the HIV era, some reports have focused specifically on musculoskeletal infections at unusual sites with unusual organisms. Other reports further document the fascinating and perplexing entity of chronic recurrent multifocal osteomyelitis. Several new treatment trends are worth noting. Acute staphylococcal osteomyelitis in many children may be better treated at reduced costs by minimizing surgical intervention, shortening the duration of antibiotic courses and hospital stays, switching quickly to the oral route for antibiotics, and not using serum bactericidal levels. Early Lyme disease, Lyme arthritis, and neuroborreliosis may respond just as well to oral doxycycline as to intravenous ceftriaxone at a much lower cost and a decreased risk of complications. PMID- 9746870 TI - [What is the value of biomechanical knowledge for surgery of the hand?]. AB - Cartilage and bone as supportive tissues are submitted to mechanical stresses and react to these in a very specific manner. Functional adaptation to the quality and magnitude of the acting forces is the result. In this article, the basic principles of functional adaptation of cartilage and bone will be presented. On the basis of these principles, promising functional concepts and clinical treatments may be established; this will be shown by some examples concerning the hand. PMID- 9746871 TI - [Changes in the form of the interosseous hood during extension and flexion of the metacarpophalangeal joint]. AB - Finger flexion initiated at the distal and proximal interphalangeal joint level forces the extensor tendon to move distally. Therefore, the interosseous hood fixed to the extensor apparatus moves distally, too. The proximal part of the hood which is located at the level of the metacarpal head during extension of the finger slides distally along the basis of the proximal phalanx. Due to the much smaller cross-section of the phalanx compared to the metacarpal head, the palmar border of the interosseous hood is shifted palmarly. This produces a two-fold effect concerning the metacarpophalangeal joint. First, it leads to an increasing flexion moment of the intrinsic muscles during flexion of the metacarpophalangeal joint. Second, the same mechanism improves the ability of abduction and adduction in the metacarpophalangeal joint during extension of the finger. In this position, the proximal part of the interosseous hood covers the metacarpal head and the strong palmar bundles of the hood are pushed to the ulnar and radial sides resulting in a greater distance to the abduction/adduction axis of the MP joint. The other effect concerns the proximal interphalangeal joint. The described transformation of the interosseous hood during flexion especially of its proximal part causes a curved deformation of the strong palmar border of the interosseous hood. The power of the intrinsic muscles inserting at the interosseous hood is passed along this smooth curve on its way to the dorsal side of the proximal interphalangeal joint thus allowing a continuous extension of the proximal interphalangeal joint in all flexion phases of the MP-joint. The typical transformation of the interosseous hood is regulated by the form of the underlying bone and ligament apparatus and can be understood as a passive mechanism effecting in a senseful change of muscle function as the active element during finger flexion and extension. These not yet described morphological data concerning the transformation of the interosseous hood during finger flexion and the functional interpretation complete the former described mechanism of flexion by Landsmeer (1955) and Landsmeer and Long (1965). PMID- 9746872 TI - [Effect of implants in the metacarpal head and proximal phalanx on mobility of the metacarpophalangeal joint. An anatomic study]. AB - Percutaneous application of Kirschner wires, fixator pins, or other implants near the metacarpophalangeal joint may lead to postoperative disability of movement with subsequent joint stiffness. To determine the best location for the application of implants into the metacarpal head and the basis of the proximal phalanx, 57 Kirschner wires were introduced into the metacarpal head and into the bases of the proximal phalanx and the resulting stiffness has been measured. Kirschner wires inserted at the level of the dorsal tubercle or proximal to the tubercle, do not lead to a noteworthy stiffness. Whenever possible, this tubercle should be taken as the distal border for inserting implants. If the position of the fracture line makes it necessary to implant wires into the metacarpal head itself, a more dorsal application is preferable. If a palmar placement cannot be avoided, the implantation should be done in neutral position of the metacarpophalangeal joint, because a transfixation of the tense collateral ligament in flexion leads to a maximum of postoperative stiffness. The application of wires into the basis of the proximal phalanx does not influence the range of movement of the metacarpophalangeal joint. PMID- 9746873 TI - [Palmar and dorsal nail anlage of the small finger. A case report]. AB - A congenital malformation of a 18-month-old boy is presented. Palmar and dorsal surface of the small finger presented a complete nail. Active flexion of the PIP and DIP joints was not possible. The small finger displayed typical dorsal skin both dorsally and palmarly. Flexion creases were absent. The palmar nail was removed, and the defect was covered by a cross-finger flap. PMID- 9746874 TI - [Bilateral congenital radio-ulnar synostosis with hyperpronation--findings and surgical therapy]. AB - Congenital proximal synostoses of the ulna and radius do not require surgery in most of the cases, since the proximal synostosis can be sufficiently compensated by the wrist joint. In cases of considerable functional deficit, the synostosis has to be separated to enable re-ossification in a more advantageous position. We report on a patient with bilateral synostosis and a significant functional deficit due to the grotesque hyperpronation of the forearms. Following operative longitudinal separation and redressive casts, a good correction was obtained. X ray imaging in lateral view documented the improved forearm position. PMID- 9746875 TI - [Pediatric scaphoid fractures--treatment and prognosis]. AB - From 1990 to 1994, we treated 28 children aged eight to fourteen years, with a scaphoid fracture. In eight of them, the initially suspected fracture could not be verified by X-ray earlier than two weeks after the injury. Eleven more patients--after false initial suspect of fracture--were discharged after two weeks without any complaints and negative X-ray control. These were not included in the series. Patients with radiologically evident fracture or clinically typical history and symptoms of scaphoid fracture--even without positive X-ray- were treated by below-elbow-thumb spica-cast for two weeks. After that period, all patients went for repeated clinical and radiological examination. All children with radiologically visible fracture or continuous, typical complaints underwent cast fixation for another four weeks. Symptom-free patients without radiological evidence of fracture were discharged. One displaced fracture was reduced and stabilized with a screw. In follow-up examination after six to forty months, all fractures showed good radiological consolidation, four patients reported occasional pain on straining the wrist. Scaphoid fractures in children seem to be more difficult to diagnose than to treat. Nondisplaced or slightly displaced fractures can be treated easily with plaster cast. Fractures that show dislocation of more than 1 mm should undergo open reduction and stabilization by a screw. PMID- 9746876 TI - [Dynamic splinting after flexor tendon injuries of the hand in childhood]. AB - Ninety severed flexor tendons were repaired in 24 boys and 14 girls in a four year-period at the Department of Pediatric Surgery in Graz. Mean age was 4.9 years. In 16 patients (42%) additional lesions were found. Direct repair was performed with modified techniques of Kirchmayr (23) or Kleinert (8). In four fingers, flexor tendon reconstructions were carried out. Postoperatively, the dynamic Kleinert splint was used for early nonresistive motion, beginning on average on the second postoperative day. Intensive training of motion was performed in physiotherapeutic sessions during a mean period of five months, postoperatively. In five cases tenolysis was performed; in one case a rupture of a sutured tendon had to be corrected. 27 patients (71%) with 42 injured fingers were followed-up on average 2.7 years after the operation. In accordance with the Buck-Gramcko classification, very good results were achieved in 37 cases (88%), and a poor result in one patient (2%). Atraumatic technique, dynamic splinting, consequent physiotherapeutic training by experienced physiotherapists, and well informed parents will yield excellent results after flexor tendon repair in the paediatric age group. PMID- 9746877 TI - [Case report of segmental neurofibromatosis of the 3rd finger ray]. AB - We report on a 36-year-old woman suffering from a segmental neurofibromatosis solely confined to the third finger. Microscopically, the tumor showed mesenchymal tissue containing myxoid material with features of the nerve sheath- myxoid neurofibroma. The tumor was characterized by the peculiar tendency to grow in a multifocal pattern involving the distribution of every nerve of one finger, by an extreme proneness to local recurrences as well as by a concomitant bone defect. PMID- 9746878 TI - [Actinic brachial plexus lesion]. AB - Radiation-induced brachial plexus lesions are progressive and irreversible complications. Until now, there is no way to successful prevention and treatment of this problem. In our series, relief of pain could be achieved by neurolysis in some cases, but there was no recovery of sensory and motor function. In order to improve the vascularity and nerve tissue regeneration, we performed muscle or gliding tissue flaps after neurolysis in our department. Since 1975, 25 patients who developed radiation-induced plexopathy were treated in our department. We followed 18 patients to evaluate the benefits of our surgical intervention. None of the patients had improvement of their sensory or motor impairment. Relief of severe pain was achieved in 83% either by neurolysis only with or without muscle or gliding tissue flap. In some cases, paresis worsened postoperatively. We also observed a return of severe pain after the operation. PMID- 9746879 TI - [Value of neurophysiological studies in diagnostic verification of carpal tunnel syndrome]. AB - The success of surgical treatment of carpal tunnel syndrome depends largely on correct diagnosis. Nerve conduction studies are usually recommended, especially in cases where history and clinical signs are not very clear. The aim of this study is to show whether nerve conduction studies are helpful in confirming the diagnosis. Fifty-two cases of carpal tunnel decompression (39 patients, 13 male, 26 female, mean age 54 years) were included in the study. Prior to surgery, all patients underwent standardized nerve conduction studies. One patient who developed RSD was excluded. In 33 cases the history was typical for carpal tunnel syndrome (Phalen 1966). The surgical result was classified excellent in 67% and good in 33%. Nerve conduction studies had failed to confirm the diagnosis in five cases, although duration as well as severity of symptoms did not differ from the other cases. In 18 cases the patients' history was considered atypical. Nerve conduction studies indicated carpal tunnel syndrome in 89%. The result of median nerve decompression was satisfactory in 78% (excellent in 45% and good in 33%) and unsatisfactory in 22% of the cases. Notably patients with unsatisfactory results had positive nerve conduction studies. The Phalen and Hoffmann-Tinel testing produced no false-positive results in these cases. General analysis showed that age, severity and duration of symptoms did not significantly influence the result. However, patients with atypical symptoms had a higher incidence of radiological evidence of cervical spondyloarthritis (50 versus 30% in patients with typical symptoms). The results of our nerve conduction studies indicated a sensitivity of 85% and a specificity of 92%, comparable favourably with other reports. We conclude that in patients with classical symptoms nerve conduction studies are not absolutely necessary. However, when symptoms are atypical the diagnosis cannot be based on nerve conduction studies alone. The false-positive results delivered by the nerve conduction studies may be explained with the "double crush" theory, since cervical spondyloarthritis prevailed in patients with atypical symptoms. PMID- 9746880 TI - [Damage pattern caused by high pressure water jets and high pressure water abrasive jets to the hand]. AB - Water and abrasive water jets have been developed as efficient tools for different purposes in industrial manufacturing, cleaning and dismantling, and- experimentally--in drilling and milling. In contrast to other high pressure techniques, these machines produce power densities up to 250 kW/mm2. In order to assess the range of destruction caused by water as well as abrasive water jets in human hands, cadaveric forelegs of pigs were targeted with constant nozzle geometry and working distance between cadaver and nozzle. Variable parameters investigated included pressure and time of exposure. Reproducible injury patterns were found in the forefoot of the pig, allowing for conclusions to be drawn with respect to damage assessment in the human hand. PMID- 9746881 TI - [Does the open palm technique for surgery of Dupuytren's contracture extend treatment and disability duration? A retrospective study]. AB - In the operative treatment of Dupuytren's contracture stage II to IV according to Iselin and Dieckmann (1951), we often prefer the open-palm technique by McCash (1964). In the past ten years, we have treated 1461 cases of Dupuytren's disease. With the assistance of a health insurance company, we followed up 214 cases. When compared to those cases treated with primary wound closure, there was no difference in regard to return to work time. PMID- 9746882 TI - [Terminology collection of radiologic concepts of modern sectional imaging methods of the hand]. PMID- 9746884 TI - Peritoneal lavage with aprotinin in patients with severe acute pancreatitis. Effects on plasma and peritoneal levels of trypsin and leukocyte proteases and their major inhibitors. AB - CONCLUSION: Although high-dose aprotinin given intraperitoneally to patients with severe acute pancreatitis seems to inhibit activated trypsin in the peritoneal cavity, the treatment has little effect on the balance between proteases and antiproteases. Plasma levels of leukocyte proteases were high in all the patients, indicating leukocyte activation to be an important feature of the pathophysiology of severe acute pancreatitis. A surprise finding was that the patients had higher peritoneal levels of pancreatic secretory trypsin inhibitor (PSTI) after the lavage procedure. BACKGROUND: Although most studies have shown protease inhibitor therapy to have little or no effect on acute pancreatitis, in an earlier study we found that very high doses of the protease inhibitor aprotinin given intraperitoneally to patients with severe acute pancreatitis seemed to reduce the need of surgical treatment for pancreatic necrosis. In the present study we have further analyzed plasma and peritoneal samples from the same patients to ascertain whether the aprotinin treatment affects the balance between proteases and endogenous antiproteases. METHODS: In a prospective double blind randomized multicenter trial, 48 patients with severe acute pancreatitis were treated with intraperitoneal lavage. One group (aprotinin group, n = 22) was also treated with high doses (20 million KIU given over 30 h) of aprotinin intraperitoneally. The remaining 26 patients made up the control group. The protease-antiprotease balance was studied by measuring immunoreactive anionic trypsin (irAT), cationic trypsin (irCT), complexes between cationic trypsin and alpha 1-protease inhibitor (irCT-alpha 1 PI), leukocyte elastase and neutrophil proteinase 4 (NP4), as well as the endogenous protease inhibitors, pancreatic secretory trypsin inhibitor (PSTI), alpha 2-macroglobulin (alpha 2M), alpha 1 protease inhibitor (alpha 1 PI), antichymotrypsin (ACHY), and secretory leukocyte protease inhibitor (SLPI). Intraperitoneal levels were studied before and after the lavage procedure, and plasma levels were followed for 21 d. RESULTS: The control group had lower plasma levels of SLPI and analysis of peritoneal fluid showed the reduction of irCT-alpha 1 PI to be more pronounced in the aprotinin group. None of the other variables measured differed significantly between the two groups. All patients had very high levels of leukocyte elastase and NP4 both in peritoneal exudate and in plasma. Peritoneal levels of PSTI were higher after the lavage procedure in contrast to the other measured variables that all showed lower peritoneal levels after the lavage. PMID- 9746883 TI - New concepts in understanding the pathophysiology of chronic pancreatitis. PMID- 9746885 TI - Oral pancreatic enzyme therapy in the control of diabetes mellitus in tropical calculous pancreatitis. AB - CONCLUSION: Pancreatic enzyme supplementation therapy conducted during a 6 mo period produces better control of diabetes, improvement in nutrition, and overall improvement in quality of life in patients with tropical calculous pancreatitis. BACKGROUND: Tropical calculous pancreatitis is characterized by abdominal pain, pancreatic calculi, and diabetes. The diabetes in insulin resistant. The brittle diabetes may be owing to defective glucagon secretion and erratic absorption of nutrients. METHODS: Patients with tropical calculous pancreatitis with diabetes and chronic pancreatitis were studied for fasting and postprandial plasma glucose, glycosylated hemoglobin, serum cholesterol, triglycerides, and calcium, liver function, and plasma C-peptide. All patients were given Creon and evaluated for quality of life. RESULTS: Clinical parameters of the patients showed considerable improvement at the end of the trial. Abdominal pain, steatorrhea, and sense of well being improved. There was a significant reduction in postprandial plasma glucose and glycosylated hemoglobin. PMID- 9746887 TI - Characteristics of pancreatic carcinoma in the elderly. AB - CONCLUSION: Lymphogenous as well as hematogenous metastases were significantly less frequent in the elderly group of patients, although local invasion was comparable. Survival was comparable between both groups although palliative therapy alone was significantly more frequent in the elderly. BACKGROUND: The relative and absolute numbers of elderly patients continue to increase, as does the incidence of pancreatic carcinoma. To determine the optimal therapy for elderly patients with pancreatic carcinoma, we examined their clinicopathological features. METHODS: The clinical and histopathological features of pancreatic carcinoma in patients 70 yr of age or older (n = 89) were compared with those in patients aged 69 yr or less (n = 184). RESULTS: A total of 273 patients showed histologically tubular adenocarcinomas and their major variants. The male:female ratio peaked at 1:0.3 in patients under 49 yr old but gradually decreased to 1:1.2 in those aged over 80 yr. There were no significant differences between the two groups in the resectability, prognosis, location, or histology of the tumor. Hematogenous and lymphogenous metastases were detected at autopsy in 68 and 61% of patients old than 70, and in 82 and 80% of the younger group. PMID- 9746886 TI - Complications requiring reoperation following pancreatectomy. AB - CONCLUSIONS: In this series, the overall reoperative rate following pancreatic surgery is 9%. Complications following pancreatectomy that require reoperation fall into four categories: hemorrhage, infectious, delayed gastric emptying, and anastomotic leak. A delay in the management of these types of complications can be fatal. BACKGROUND: Despite the improvement in the morbidity and mortality rates associated with pancreatic resection, complication still arise that require surgical intervention. This study reviews the pancreatic surgical experience at a major medical center to determine the overall reoperative complication rate. STUDY DESIGN: From 1985 to 1995, 107 patients underwent pancreatic resection. There were 50 pancreaticoduodenectomies, 20 total pancreatectomies, and 37 distal pancreatectomies for 102 periampullary or pancreatic cancers and five for chronic pancreatitis. The operative mortality rate was 6.5% and the morbidity rate was 43%. Ten patients (9%) developed complications that required reoperation. RESULTS: Re-exploration was performed in five patients for hemorrhage. Four patients had bleeding intra-abdominally and one had a suture line bleed. One patient developed a wound infection and fascial necrosis which necessitated reoperation. Three patients were explored for sepsis and one was found to have a pancreatic leak. One patient had persistent gastric outlet obstruction and he required conversion of the gastrojejunostomy to a Roux-en-y anastomosis. The mortality rate for re-exploration was 3/10 (30%). PMID- 9746889 TI - Primary hyperparathyroidism and acute pancreatitis during pregnancy. Report of a case and a review of the English and Japanese literature. AB - The simultaneous occurrence of primary hyperparathyroidism (PHPT) and pancreatitis during pregnancy is very rare. We present a case of concurrent PHPT and pancreatitis in pregnancy and review 13 cases reported in the English and Japanese literature. Two maternal and three fetal deaths occurred. Mortality seemed to be related to delayed resection of the parathyroid tumor. Morphologically, severe pancreatitis was only seen in three cases, whereas even edematous or focal pancreatitis caused the same symptoms as clinically severe pancreatitis. Acute pancreatitis should be kept in mind in the differential diagnosis of unexplained nausea and abdominal pain during pregnancy, and hyper-or normocalcemia in the severe form of pancreatitis should be a clue to concurrent PHPT. PMID- 9746888 TI - Effects of epidermal growth factor on neonatal pancreatic growth in the guinea pig. AB - CONCLUSION: EGF and/or transforming growth factor-alpha (TGF-alpha) are likely to be important in the rapid pancreatic growth that occurs in the neonatal guinea pig. BACKGROUND: Rapid pancreatic growth is observed during the neonatal period in the guinea pig. The growth factors that are involved are not known but may include members of the EGF family. METHODS: Mini-osmotic pumps were implanted on the day of birth for continuous infusion of EGF (30 microgram/d). Pancreatic DNA, RNA, and protein contents were determined at 4 and 15 d, along with wet weights of the pancreas, duodenum, jejunoileum, colon, and gallbladder. Pancreatic EGF and TGF-alpha concentrations were measured in adult controls, in control neonates at 1, 4, 8, and 15 d, and also at 4 and 15 in guinea pigs receiving either EGF or the cholecystokinin receptor antagonist devazepide (25 nmol/kg/h). RESULTS: EGF infusion significantly increased the weight of the stomach and duodenum at 4 d and all the gastrointestinal organs, including the pancreas, at 15 d. Exogenous EGF increased pancreatic DNA, RNA, and protein content at 4 and 15 d. Endogenous EGF and TGF-alpha concentrations in the pancreas were significantly higher at birth than in adults (P < 0.001) and P < 0.01, respectively) and declined during the first 2 wk postpartum. At 15 d, EGF concentrations remained significantly higher than adult levels (P < 0.01), but TGF-alpha concentrations had declined to adult values. Infusion of EGF decreased concentrations of endogenous EGF in the pancreas at 4 and 15 d (both P < 0.05) and decreased TGF-alpha concentrations at 4 d (P < 0.001). Devazepide infusion caused a significant decrease in endogenous pancreatic EGF concentrations at 15 d (P < 0.05). PMID- 9746890 TI - Indications for surgical resection of metastatic ocular melanoma. A case report and review of the literature. AB - CONCLUSIONS: Ocular melanoma can metastasize to the gallbladder and porta hepatic nodes and mimic pancreatic carcinoma. If one suspects metastatic disease, a complete metastatic work-up must be done prior to surgery to prevent unnecessary surgery. If no distant disease is present or the patient is symptomatic, metastatic disease should be resected. PURPOSE: To review the literature pertaining to the spread of ocular melanoma and to determine if distant disease should be resected. PATIENTS AND METHODS: A 44-yr-old Egyptian male presented to an outside institution with mid-epigastric and right upper quadrant abdominal pain. His past medical history was significant for a left orbital enucleation for uveal melanoma in 1982. On physical examination, there was no supraclavicular adenopathy and no skin lesions were noted. There was a mass in the right upper quadrant. The total bilirubin was 4.8 mg/dL. A computed tomography showed a mass in the head of the pancreas and portal vein involvement could not be determined. RESULTS: The patient was taken to the operating room and a pancreatico duodenectomy was performed for a cystic mass in the head of the pancreas. Final pathology revealed metastatic melanoma in the gallbladder and an enlarged, cystic lymph node growing into the head of the pancreas replaced with metastatic melanoma. The patient did well post-operatively and was discharged home on the eighth post-operative day. PMID- 9746891 TI - Pancreatic metastasis from a lung cancer. Preoperative diagnosis and management. AB - A case of a surgically resectable solitary metastasis to the pancreas from a lung cancer, confirmed by immunohistochemical staining (PE-10), is reported. ERCP revealed meniscoid interruption of the main pancreatic duct, which is uncommon in patients with primary pancreatic cancers of the pancreas in our hospital. This patient lived for 29 mo after the surgical resection of the pancreatic lesion. Therefore when metastasis limited to the pancreas is evident on clinical imaging, surgical management may be more optimal than other treatment approaches. PMID- 9746892 TI - Quality of life assessment in chronic pancreatitis. PMID- 9746893 TI - EORTC QLQ-30. A breach of copyright. PMID- 9746894 TI - Diverse rationalities and multiple realities in illness and healing. PMID- 9746895 TI - In search of the good: narrative reasoning in clinical practice. AB - Based on ethnographic work among North American occupational therapists, I compare two forms of everyday clinical talk. One, "chart talk," conforms to normative conceptions of clinical rationality. The second, storytelling, permeates clinical discussions but has no formal status as a vehicle for clinical reasoning. I argue that both modes of discourse provide avenues for reasoning about clinical problems. However, these discourses construct very different clinical objects and different phenomena to reason about. Further, the clinical problems created through storytelling point toward a more radically distinct conception of rationality than the one underlying biomedicine as it is formally conceived. Clinical storytelling is more usefully understood as a mode of Aristotle's "practical rationality" than the technical rationality of modern (enlightenment) conceptions of reasoning. PMID- 9746896 TI - Moral reasoning and the meaning of cancer: causal explanations of oncologists and patients in southern Mexico. AB - Moral themes were a striking feature of the causal explanations for female cancers discussed by oncologists and patients in an ethnographic study of hospital-based cancer care in southern Mexico. These explanations integrate general biomedical explanations with everyday expectations and experiences, giving meaning to otherwise arbitrary events. Analysis of case examples shows that causal models incorporate local constructs about what constitutes a virtuous life, especially in terms of class-and gender-based values. Although patients and physicians draw on similar concepts of moral order, they apply these constructs in distinct ways. Because physicians' explanations are necessarily framed in terms of object, their causal stories employ generalized presumptions about how categories of persons behave (e.g., women, the lower class). In contrast, patients' explanations are framed in terms of subject; they are based on the specific details of their personal history. The article examines the distinct perspectives of physicians and patients, and provides an illustration of how biomedical culture articulates with the local moral constructs of a particular community. PMID- 9746897 TI - On the rationality of decision-making studies: Part 1: Decision models of treatment choice. AB - With reference to both critiques and empirical studies, the theoretical and methodological grounding of anthropological research on medical decision making is examined in this article, giving particular attention to the construction and evaluation of cognitively oriented decision models. A decision-modeling study carried out in the Mexican village of Pichataro (in conjunction with James C. Young) frames an exploration of some of the tensions and points of contention about the aims and designs of cognitively oriented studies of decision modeling. While a decision model can provide a reasonably good guide to an understanding of treatment actions and the culturally based rationality that underlies them, such models fall short when they are oriented primarily around predicting treatment accounts. They should also attend to the jointly cultural, personal, social, and cognitive constructive processes through which meaning is conferred upon the occurrence of illness. PMID- 9746898 TI - On the rationality of decision-making studies: Part 2: Divergent rationalities. AB - A study of treatment decision making in an Anishinaabe community in Manitoba, Canada was designed to be comparable with an earlier project carried out in a Mexican town. One objective was to compare the resulting decision models. For both communities, a decision-making perspective was compatible with how individuals talked about actions taken in response to illness, and it proved to be a useful means for learning about the process of seeking care. At the same time, a decision-modeling approach is better suited to explaining treatment actions taken in the Mexican community than in the Anishinaabe community. I suggest that this finding reflects the variable potentiality, in the Anishinaabe community, for affliction and its treatment to be constructed within a cultural framework in which the underlying assumptions differ from those implicit in studies of decision modeling. PMID- 9746899 TI - Varieties of reasoning in medical anthropology. PMID- 9746900 TI - Methodological approaches to the study of reasoning. PMID- 9746901 TI - Repairing broken rules: care-seeking narratives for menstrual problems in rural Mali. AB - Narratives play an important role in the organization of therapeutic action in rural Mali. This article provides structural and interpretive analyses of a young, French-speaking Dogon woman's accounts of her efforts to manage her menstrual bleeding and threatened infertility. Through her personal narratives she creates social arenas to recruit support, negotiate changes in her family relationships, and enhance her standing as a member of the community. Beginning with the accounts of her fear and helplessness, the narrator integrates past events into her unfolding present and achieves a meaningful resolution of her problem. Her narratives weave together encounters with family members, friends, and healers to describe a therapeutic itinerary that acquires significance as a transformative experience. PMID- 9746902 TI - [Tolcapon potentiates the Levodopa effect]. PMID- 9746903 TI - [Current pathogenetic and immunologic aspects of multiple sclerosis]. PMID- 9746904 TI - Modulating excitatory synaptic neurotransmission: potential treatment for neurological disease? AB - Excitatory neurotransmission at many CNS synapses depends upon AMPA-type glutamate receptors. Derangements in AMPA receptor-mediated synaptic transmission may be a contributing factor in neurological and neurodegenerative diseases and could be a target for therapeutic intervention. Drugs that positively modulate AMPA receptors by reducing AMPA receptor desensitization and/or slowing AMPA receptor deactivation, such as thiazide derivative (cyclothiazide, diazoxide, IDRA 21) and benzoylpiperidine derivatives (1-BCP, CX516, aniracetam), facilitate AMPA receptor-mediated processes and may have beneficial therapeutic effects. For example, AMPA modulators facilitate long-term potentiation, which may be important for memory storage, and facilitate memory encoding in behavioral experiments. Thus, AMPA modulators might ameliorate memory deficits that occur in dementia, such as Alzheimer's disease. However, AMPA receptor-mediated excitotoxicity may occur with excessive AMPA receptor activation such as in seizures or ischemia, and positive AMPA modulators would promote neuronal injury under those conditions. Regardless of the ultimate clinical utility of positive AMPA modulators, their discovery and study have already provided significant insight into the physiology and structural determinants of important AMPA receptor properties. This review attempts to synthesize a variety of studies that have utilized these AMPA modulators to gain insight into fundamental as well as clinically relevant AMPA receptor-mediated processes. PMID- 9746905 TI - Enhancement of outward potassium current may participate in beta-amyloid peptide induced cortical neuronal death. AB - In light of recent evidence implicating the upregulation of outward K+ current in mediating several forms of neuronal apoptosis, we tested the hypothesis that such an upregulation might specifically contribute to the pathogenesis of beta-amyloid peptide (A beta)-induced neuronal death. Exposure to A beta fragment 25-35 (20 microM) or 1-42 (20 microM) enhanced the delayed rectifier K+ current IK, shifting its activation voltage relationship toward hyperpolarized levels and increasing maximal conductance, but did not affect the transient K+ current IA or charybdotoxin-sensitive BK current. Reducing IK by adding the channel blocker tetraethylammonium (TEA, 5 mM) or raising extracellular K+ to 25 mM attenuated A beta-induced neuronal death, even in the presence of nifedipine or gadolinium to block associated increases in Ca2+ influx. The IA blocker 4-aminopyridine (4-AP, 5 mM) and the CI- channel blocker anthracene-9-carboxylic acid (ACA, 500 microM) were not neuroprotective. These data raise the intriguing possibility that manipulations aimed at reducing outward K+ current may provide an approach to reducing neuronal degeneration in patients with Alzheimer's disease. PMID- 9746906 TI - Identification of a familial mutation associated with GABA-transaminase deficiency disease. AB - GABA-transaminase (GABA-T) deficiency disease is a rare recessive disorder characterized by abnormal development, seizures, and high levels of GABA in serum and cerebrospinal fluid. Although some patients are the offspring of consanguineous marriages, most are not. To identify the molecular basis of this disease, we have determined the sequence of human GABA-T cDNA. We have compared the GABA-T cDNA sequences in cultured cells derived from six healthy controls with those from a GABA-T-deficient patient and both parents. Our data indicate that GABA-T deficiency disease may result from an allele that encodes an R220K substitution. PMID- 9746907 TI - A novel immunophilin ligand: distinct branching effects on dopaminergic neurons in culture and neurotrophic actions after oral administration in an animal model of Parkinson's disease. AB - Protection or regeneration of the dopaminergic (DA) system would be of significant therapeutic value for Parkinson's disease. Immunophilin ligands, such as FK506, can produce neurotrophic effects in vitro and in vivo, but their immunosuppressive effects make them unsuitable for neurological application. This study demonstrates that a novel, nonimmunosuppressive immunophilin ligand (V 10,367) increased the number of neurites extended by tyrosine hydroxylase positive (TH+) DA neurons in embryonic day 14 primary DA neuronal cultures. In contrast, the immunosuppressive immunophilin ligand FK506 increased the length of TH+ neurites. After oral administration in MPTP-treated mice, V-10,367 completely protected against MPTP-induced loss of striatal TH+ axonal density, while FK506 did not. These experiments demonstrate that nonimmunosuppressive immunophilin ligands specifically increase neurite branching in primary DA neuronal culture and possess neurotrophic actions in vivo with potential application to neurodegenerative disease. PMID- 9746908 TI - Additive effects of PS1 and APP mutations on secretion of the 42-residue amyloid beta-protein. AB - Humans harboring missense mutations in the presenilin 1 (PS1) gene undergo progressive cerebral deposition of the 42-residue amyloid beta-protein (A beta 42) at an early age and develop severe Alzheimer's disease. A beta 42 is selectively elevated in the conditioned media of cells expressing mutant but not wild-type PS1, indicating that presenilin mutations alter APP processing. Here we analyze the effects of various PS1 mutant constructs on the cellular production of A beta 42. A construct expressing only the PS1 N-terminal endoproteolytic fragment with the mutation Y115H causes no significant increase in A beta 42, whereas a full-length PS1 construct with the same mutation does. This result suggests that the pathogenic effect of mutant presenilins is produced by the full length molecule even though only a minor proportion of total PS1 occurs as holoprotein in tissues and cell lines. We demonstrate that the effects of two different PS1 mutations are additive when engineered into the same PS1 molecule. Therefore, two mutations alter gamma-secretase processing of APP more than one and the PS1 mutations described to date do not cause the maximum possible PS1 mediated rise in A beta 42. When a PS1 mutation was expressed in cells carrying the APPV717I mutation, A beta 42 rose dramatically to become the predominant secreted A beta species, an observation of interest for transgenic modeling of AD. Our results are consistent with the hypothesis that presenilin is a major regulator of the proteolytic processing of APP by gamma-secretases. PMID- 9746909 TI - Effects of streptozotocin-induced hyperglycemia on brain damage following transient ischemia. AB - Hyperglycemia is known to aggravate ischemic brain damage. The present experiments were undertaken to explore whether hyperglycemia caused by streptozotocin-induced diabetes exacerbates brain damage following transient brain ischemia as it does in animals acutely infused with glucose. Experimental diabetes was induced by injection of streptozotocin in rats which were subjected to 10 min of forebrain ischemia either 1 week (1-wk) or 4 weeks (4-wk) after the induction of diabetes. Normoglycemic rats exposed to the same duration of ischemia and sham-operated diabetic rats served as controls. The animals underwent evaluation of clinical outcome and histopathological analysis of brain damage. Postischemic seizures developed in 35.3 and 42.1% of 1-wk and 4-wk diabetic hyperglycemic animals, respectively. The incidence of seizure was not different between the two groups. None of the diabetic animals with plasma glucose concentrations below 12 mM exhibited seizure activity. The extent and distribution of brain damage were similar between 1-and 4-wk diabetic animals. In the CA1 and in the subicular regions of hippocampus, both diabetic hyperglycemic and normoglycemic animals showed 70-80% cell death. Diabetic hyperglycemic animals had more severe neuronal necrosis in the parietal cortex than normoglycemic animals. In diabetic hyperglycemic animals, neuronal damage involved additional brain structures, e.g., cingulate cortex, thalamus nuclei, substantia nigra, pars reticulata, and the hippocampal CA3 sector, i.e., structures in which neurons were not affected in normoglycemic ischemic subjects at this duration of ischemia. These findings demonstrate that diabetic hyperglycemic animals frequently develop postischemic seizures and that streptozotocin-induced hyperglycemia results exacerbated postischemic brain damage of the same density and distribution as in acutely glucose-infused animals. PMID- 9746910 TI - [Grading of astrocytomas and oligodendrogliomas]. AB - The histologic determination of the degree of tissue anaplasia and grade of malignancy of gliomas is based upon qualitative histological features (nuclear pleomorphism, mitoses, endothelial proliferation, tumor necrosis). This grading approach is influenced by the subjective interpretation of the pathologist, especially concerning the weighting of criteria (scant, moderate, pronounced). An observer-independent approach seems to be feasible by abandoning the concept of parameter weighting in favor of an binary approach noting only the presence or absence of these structure parameters. This grading procedure is recognized in the revised WHO classification of brain tumors for common type astrocytomas (Ste. Anne-Mayo System, SAMS). Our results indicate that a similar approach is also suitable for grading purposes of oligodendrogliomas and mixed gliomas. Our recent investigations on glioma grading showed, both for astrocytomas and oligodendrogliomas, that a two-tiered grading scheme distinguishing only "low grade" and "high grade" cases was prognostically relevant. For all glioma entities the onset of tumor angiogenesis with endothelial proliferation and contrast enhancement in CT and MRI seems to be the key criterion indicating irreversible tumor progression to the "high" malignancy grade. PMID- 9746911 TI - [Molecular cancer disposition diagnosis exemplified by colorectal carcinoma. What is the contribution of pathology?]. AB - During the last few years, the molecular basis of several cancer predisposition syndromes has been discovered which offers new tools for cancer prevention and early detection. This will be demonstrated in one of the most frequent hereditary cancer syndromes, namely the hereditary nonpolyposis colorectal cancer (HNPCC) which accounts for about 5% to 8% of CRC. Thereby, families with exclusively CRC (Lynch type I syndrome) and those with extracolonic cancers especially of endometrium, stomach, small bowel and upper urinary tract (Lynch type II syndrome) can be discriminated. At the molecular level, HNPCC is caused by germline mutations in one of the mismatch repair genes (hMSH2, hMLH1, hMSH6, hPMS2). Thus, nucleotide mispairings occurring particularly within simple repetitive genomic sequences (microsatellites) during replication are no longer be repaired properly and can be demonstrated by PCR as so-called microsatellite instability (MSI). Since more than 90% of HNPCC associated and only about 15% of sporadic CRC show MSI, this test is a useful tool for HNPCC screening. In case of a negative result HNPCC is highly unlikely. In positive cases (with > or = 2 out of 5 unstable defined microsatellite markers) the definite molecular diagnosis can only be obtained by sequencing the mismatch repair genes from the patient's blood or normal DNA. As immunohistochemistry reveals loss of hMSH2 or hMLH1 expression in most MSI positive CRC, these data provide useful information for the sequencing strategy. Molecular tumor screening by MSI test and immunohistochemistry is recommended in patients i.) with a positive family history (acc. to the Amsterdam criteria), ii.) suffering from multiple HNPCC related carcinomas, iii.) with HNPCC related cancer before 45 ys of age, and iv.) with right-sided CRC exhibiting medullary, signet-ring or mucinous differentiation. Finally, these tests as well as genetic counseling and treatment of the patient need to be done by an interdisciplinary approach. Thereby, the pathologist can substantially contribute to identify HNPCC related carcinomas either by clinical or morphological criteria and to initiate the molecular screening test. PMID- 9746912 TI - [Pattern of various cytokeratins of normal vulva, vulvar intraepithelial neoplasia (VIN) and vulvar carcinoma]. AB - The presence of specific keratins can be of diagnostic value for studying normal and neoplastic epithelium of the vulva. The aim of the present study was to investigate normal, preneoplastic and neoplastic epithelium of the vulva. Keratins 5, 6 and 18, identified by a polyspecific anti-human CK antibody (clone LP 34, DAKO), and the keratin subtypes 7, 10, 14, 18, 19 and 20 of normal, dysplastic and malignant vulval epithelium (paraffin-embedded sections) were detected by immunohistochemical APAAP staining. Keratins 5, 6, and 18 (clone LP 34) and keratin subtype 10 are expressed in the upper third of the normal vulval epithelium. In mild and moderate intraepithelial neoplasia only a few cells express these keratins. In patients with severe intraepithelial neoplasia (VIN III) the expression of these keratins seems to be associated with recurrence of the disease. In biopsy specimens of patients without recurrence we find positive results for keratins 5, 6 and 18 (clone LP 34) and keratin 10. If patients have a recurrence of the disease, expression of these keratins is only diffuse or is absent. The expression of these keratin subtypes in vulval carcinomas is mostly seen in differentiated cells. There was no association between recurrence and keratin pattern. We have not found any other expression of the tested keratin subtypes in VIN and in vulval carcinoma. PMID- 9746913 TI - [Juvenile pleomorphic parotid adenoma of embryonal structure]. AB - Juvenile pleomorphic adenoma of the parotid gland represents an extremely rare tumour entity and is comparable to congenital tumours of the salivary glands concerning its embryonal structure. The clinical detection of the tumour in a 7 year-old girl does not exclude that the tumour had developed either earlier or immediately after the birth. The high cellularity and the evidence of primitive epithelial and myoepithelial cellular structures do not justify its classification as a malignant tumour. However the presence of embryonal tissue structures associated with the end of the third month of embryogenesis is characterized by more solid cell formations in partly verticulate arrangement. The absence of further differentiation into lobular structures and differentiated duct or acinic cell formations may be due to cell arrest. Differential diagnosis of juvenile pleomorphic adenoma must distinguished it from other congenital salivary gland tumours (e.g. congenital basal cell adenoma, hybrid basal cell adenoma-adenoid cystic carcinoma, sialoblastoma, salivary gland anlage tumour. PMID- 9746914 TI - [Acquired, cystic kidney disease in chronic dialysis patients: a retrospective study of 125 autopsies. 1: Cysts]. AB - In acquired cystic kidney disease cysts develop in kidneys with impaired excretory function. Bleeding, cyst infection and a possible relation to kidney cell carcinoma resulted in an interdisciplinary concern about this disease. Kidney tissue slides from 125 autopsies of dialysis patients were studied. Of totally 967 cysts, 52% were lined by a cuboidal epithelium, with either clear or eosinophilic cytoplasm; 34% showed a flat epithelium. Multilayered epithelium was present in 8.3% and bleeding stigmata in 5.3%. Cyst counts rose with duration of dialysis, with a higher average cyst count in men. There was no correlation between cysts and (1) age, at the time of death of at the beginning of dialysis, (2) type of renal disease, (3) blood group, (4) body weight, (5) coronary heart disease, and (6) diabetes mellitus. More studies on the development of cysts, parameters of cyst formation and complications are necessary, as they are the basis for the interpretation of new imaging techniques, as well as, for a clinical risk assessment. PMID- 9746915 TI - [Primary bronchial malignant melanoma]. AB - A 53-year-old man presented a melanotic lung tumor which was based in the bronchus of the left lower lobe and closed the left main bronchus. After laser therapy, left lobectomy with sleeve resection was carried out. Complications after the surgery required resection of the rest of the left lung and thoracoplasty. Based on the histological and immunohistochemical findings, the tumor was classified as a malignant melanoma. There was no past history of an excision or a fulguration of a cutaneous, mucous membrane, or ocular lesion. Examination of the skin and the eyes did not yield any evidence of another primary tumor. We conclude that the lesion represents a primary malignant melanoma of the respiratory tract, a rare neoplasm of which only 21 cases have been confirmed. The patient does not have any evidence of tumor in the relatively short follow-up period of 10 months. PMID- 9746917 TI - [Myositis proliferans: a pseudosarcomatous lesion in soft tissue]. AB - Myositis proliferans is a reactive, intramuscular soft tissue disease characterized by fibroblast and myofibroblast proliferation, showing similarities to the phase-like development seen in the general pathology of wound healing and hypertrophic scars. Immunohistochemically, a combined expression of vimentin and alpha-sm actin is seen in the spindle-shaped cell formations. The decisive histological preparations is supported by immunohistochemical techniques, especially in the differentiation from sarcoma. If a definite diagnosis is made, incomplete excision may suffice. PMID- 9746916 TI - [Nevus cell aggregates in axillary lymph nodes in simultaneous mucinous breast carcinoma]. AB - Nevus cell aggregates in lymph nodes are uncommon. This benign phenomenon may be difficult to differentiate from metastatic neoplasia. We report the case of a 56 year-old patient who underwent breast biopsy, followed by radical mastectomy including lymphadenectomy. Histological examination revealed solid cell aggregates as foreign tissue in the capsule of 1 of 11 identified lymph nodes devoid of any keratin immunoreaction. Strong immunohistological staining for the S-100 protein confirmed the diagnosis of nevus cell aggregates. PMID- 9746918 TI - [In situ PCR and PCR in situ hybridization of paraffin-embedded tissue. New diagnostic possibilities in pathology]. AB - In situ polymerase chain reaction (PCR) and PCR in situ hybridization are new diagnostic tools in dermatopathology. These techniques can combine the sensitivity of PCR with the advantage of in situ hybridization to localize specific cellular structures. With optical control of the PCR reaction product in these techniques, false-positive results due to contamination can be minimized. Several working protocols have been established which allow detection of DNA and RNA sequences in a rapid and reproducible manner. Two different methods have been established to detect the amplification product, the indirect PCR in situ hybridization (PCRisHyb), and the direct in situ PCR (isPCR). An easy and reproducible method for PCRisHyb and isPCR in formalin-fixed and paraffin embedded tissue is described and the two techniques are compared. Using both isPCR and PCRisHyb, the amplification product can be visualized with an optimal morphological preservation of the tissue. Indirect PCRisHyb showed a slightly higher specificity whereas direct isPCR was the quicker, easier and less expensive method. PMID- 9746920 TI - [Meningeal melanocytoma]. AB - A 38-year-old man presented with an intracranial extra-axial tumour at the base of the left posterior fossa which proved to be a meningeal melanocytoma. Meningeal melanocytoma is a rare, benign melanotic tumour of the leptomeninges occurring predominantly in the posterior fossa or the upper spinal cord in adults. It shows characteristic cytologic features with isomorphic epitheliod or spindle-shaped cells, often with prominent nucleoli and a variable content of intracytoplasmic melanin. It usually lacks signs of malignancy such as high mitotic rate, necroses or infiltrative growth and shows a low labeling index in proliferation marker studies. Its immunohistological profile with S-100 protein-, vimentin- and HMB-45-positive tumour cells is similar to that of (primary or metastatic) malignant melanoma. This differential diagnosis is crucial because of the totally different therapeutic and prognostic implications. Therefore, everyone dealing with surgical neuropathology should be familiar with the rare, but clinically important diagnosis of meningeal melanocytoma. PMID- 9746919 TI - [Economic evaluation of telepathology]. AB - Telepathology, defined as the practice of pathology over a distance by viewing images transmitted from a remote site, was previously analyzed mainly with respect to technical equipment and diagnostic accuracy compared with conventional histological diagnosis. In this paper, telepathology is analyzed with regard to economic points of view, demonstrating the background for the cost-saving analysis of telepathology. The acquisition cost and the operating costs of a telepathology system are compared with potential cost-saving and other benefits from telepathology. This analysis may help to decide whether or not telepathology should be established in a hospital. PMID- 9746921 TI - [Helicobacter pylori and stomach carcinoma. Is a preventive intervention study of value?]. PMID- 9746922 TI - [Persistent udder edema as a herd problem: a literature review]. AB - The data presented in literature suggest that disturbances of volume regulation do not have a role in the pathogenesis of persistent udder oedema. Instead, they favour local mammary problems. The presence of both mast cells and oedema, especially in subcutaneous tissue, focuses attention on the role of mast cells in the pathogenesis of persistent udder oedema via increased endothelial permeability due to histamine release. The dissociation constants (Kd) for the high-affinity glucocorticoid receptor of normal and oedematous udders are not significantly different, and nor are there differences between the total quantity of dexamethasone bound (Bmax) and mast cell density in normal and oedematous udders. It has been shown that, in cattle, the concentration of histamine is higher in milk than in plasma. In addition, the urine histamine concentration is affected by the diet. Treatments to reduce udder oedema include combined salt and water restriction, supplementation with potassium or vitamin E (alpha-tocopheryl acetate), sodium restriction, and administration of hydrochlorothiazide intramuscularly, chlorothiazide orally, or sodium-acetazolamide orally and/or parenterally (i.v./i.m.). A low cation-anion balance in a diet containing 1.6% calcium also seems to reduce mammary oedema formation. PMID- 9746924 TI - [Equine reproduction: a much-enlarged ovary]. PMID- 9746923 TI - [Bacteriologic detection and sensitivity determination combined: a rapid test for use in practice?]. AB - In this investigation the results obtained with a testkit for detecting bacteria and their antibiotic sensitivity (ABcheck, Vetoquinol) were compared with those obtained using standard bacteriological procedures. Ten urine samples, ten skin samples, and ten cerumen samples were examined in parallel. The results indicate that the testkit is not reliable with respect to whether antibiotics should be administered and with respect to which antibiotics should be used. PMID- 9746925 TI - [Predecessors: veterinarians from earlier times. 29. William Dick (1793-1866)]. PMID- 9746926 TI - [Nutrition of dogs and cats has scientific interest. IAMS Nutrition Symposium 1998]. PMID- 9746927 TI - ["Homeopathy opening up." Homeopathy under guarantees of quality and disability]. PMID- 9746930 TI - [The political power of elders: an illusion or a reality? A gap in science]. PMID- 9746931 TI - [Older chronic patients and their care provisions in the first half of the twentieth century]. AB - Contrary to popular beliefs the Dutch nursing home came into existence not after, but long before 1945. The special care units for the old chronic patient (developed between 1900 and 1950) were a response to the problematic status of these patients in the workhouses and general hospitals. In both institutions, they occupied 'the wrong bed'. The workhouses were subject to a lot of criticism because the provided housing for the complete range of elderly persons. This criticism triggered a growing need for specialized and separated housing facilities for the able and disabled elderly. In general hospitals the elderly patients occupied beds needed for other patients, while the care was too expensive. This problem increased because of the ageing of the hospital population. The latter was caused by the age-related increase of both the admission ratio and the average length of stay. In reaction to these problems, inside and outside the existing facilities, special care units with emphasis on nursing, were developed for the elderly chronic patient. Thus, during the first half of this century this provided the foundation of the future nursing homes, which developed so successfully after 1950. PMID- 9746932 TI - [Are active neurons a better defense against aging in Alzheimer's disease?]. AB - This article deals with the question whether metabolic activity of neurons interferes with their survival during brain aging and Alzheimer's disease (AD). This 'use it or lose it' concept assumes that active neurons have a better chance to survive these conditions. We have monitored activity changes in human hypothalamic nuclei, that show differential survival patterns in aging and AD. The size of the Golgi apparatus (GA) was measured in e.g. the nucleus basalis of Meynert (NBM), that is severely affected in AD, and in the vasopressin (AVP) containing neurons of the supraoptic nucleus (SON) that remain very stable and show no cell loss. In the affected NBM, a strong reduction in activity was found in AD, whereas in the stable SON, an increased activity was present in both conditions. These findings agree with the concept that activation is associated with pronounced stability in aging and AD. Another hypothalamic nucleus is the biological clock (SCN), which is very sensitive to light input. It loses about 35% of its AVP cells in old rats. In order to test the hypothesis that extra stimulation prevents degeneration, the SCN in old rats was activated by means of an increased light input. This could indeed prevent the age-related loss of AVP neurons in the SCN in low light conditions. Increased light also restored the age related decreased amplitude in the sleep-wake rhythm. Furthermore, in AD patients, increased amounts of environmental light improved day-night rhythms and reduced behavioural disturbances. These observations are in line with the 'use it or lose it' concept. Furthermore, oxidative damage to the DNA was studied as a) it may accumulate during neuronal aging, and b) activated cells repair their DNA more efficiently. Whereas biochemical measurements of 8OHDG levels were not different in aging or AD, in situ end labeling, that detects fragmented DNA histologically, showed many positive neurons and glial cells in the AD, but not control, hippocampus, whereas in SON and PVN, hardly any damage was detected, which agrees with the 'use it or lose it' concept. Supported by related literature, we conclude that activation may be effective for neuronal maintenance during aging and in AD, and may provide a fruitful basis in the search for future treatment strategies in AD. PMID- 9746933 TI - [Aging of the crystalline lens and presbyopia]. AB - The crystalline lens changes both optically and mechanically with age: it becomes less transparent and more difficult to deform. Using a new ultrasound technique, Continuous Ultrasonographic Biometry (CUB), we were able to demonstrate that presbyopia is caused by increased stiffness of the lens due to changes in lens fibre membranes and cytoskeleton. These results are important for further research on the ageing processes that lead to cataract and presbyopia, and the treatment of these conditions. PMID- 9746934 TI - [Cognitive deficits in Parkinson's disease]. AB - In neuropsychological studies of Parkinson's disease, cognitive deficits are frequently reported, but the nature of these deficits is not clear. As far as cognitive deficits are manifest in parkinsonian patients at an early stage of the disease, many studies tend to describe them as fitting a frontal syndrome. As a consequence of dysfunction of the striatum, the (pre)frontal cortex receives deficient input from the striatum, which might explain the similarity of the cognitive deficits of parkinsonian patients with those of patients with frontal dysfunction. The present studies provide evidence that the cognitive deficits of parkinsonian patients display a certain similarity with those of patients with frontal dysfunction at the level of the ultimate performance, but that the underlying processes have a distinct character. Parkinsonian patients exhibit a decrease in self-generated problem-solving. This deficit is manifest at a level of cognitive function, which goes beyond task or domain. Among all disease variables, only severity of the disease and especially rigidity proved to be related consistently to this decrease in self-generated problem-solving. PMID- 9746935 TI - [Future time perspectives of the elderly; an empirical study rooted in theory]. AB - Which factors other than age play a role in the future time perspective of elderly people? In the literature no consensus exists on these factors. Besides, the usual approach of future time perspective as 'extension' seems limited and less suited for elderly people. Therefore, in this study the future time perspective is defined and measured as the expectation of the future with respect to new or enduring possibilities (a positive perspective) or increasing restrictions (a negative perspective). The role of 'a sense of control'--by some authors emphasized as an important factor in the future time perspective--is elaborated under the concept of 'competence belief'. Based on a theoretical framework, an elaboration of the antecedents of the future time perspective of elderly people is suggested. This led to specific expectations, which are empirically tested in a group of 4792 elderly of age 57 years and over. The results show that age indeed has an in-fluence on the future time perspective of elderly, but resources play a relatively larger role than age. This holds especially for vitality, social contact and affection, and to a lesser degree for income and activity. Also the expected positive relationship between competence belief was controlled for. The main conclusion is that elderly people show a more positive view towards the future, the more vitality, social contacts and affection they have, and the more they believe themselves associated to be competent. PMID- 9746936 TI - Bacteriophage biology and bacterial virulence. PMID- 9746937 TI - Identification of a competence regulon in Streptococcus pneumoniae by genomic analysis. PMID- 9746938 TI - The INT1 of Candida albicans. PMID- 9746939 TI - Pathogenesis of chronic bacterial infections. PMID- 9746940 TI - Structure, function and stability of enzymes from the Archaea. AB - The Archaea include microorganisms growing in some of the most extreme environments on earth. Consequently, their cellular components are remarkably stable entities and have considerable potential in the biotechnology industry. Here, we review the structure of archaeal enzymes in the context of their ability to function at extremes of temperature, salinity, pH and pressure. PMID- 9746941 TI - Starting up yeast glycolysis. AB - Successfully igniting the yeast glycolytic flux during the transition from gluconeogenic to fermentative growth seems to be a matter of balance and coordination between a multitude of events. The contours of the sugar sensing and signalling pathways that regulate this transition are only beginning to emerge. PMID- 9746942 TI - Segregation of minichromosomes in trypanosomes: implications for mitotic mechanisms. AB - In addition to 11 pairs of housekeeping chromosomes, the genome of Trypanosoma brucei contains approximately 100 minichromosomes that are probably involved in the ability of the parasite to evade the host's immune response. This minichromosomal population is segregated on the mitotic spindle. How this is achieved provides insight into potential segregation mechanisms for small DNA molecules in eukaryotic microorganisms. PMID- 9746943 TI - beta-Lactamases: protein evolution in real time. AB - The evolution and spread of bacteria resistant to beta-lactam antibiotics has progressed at an alarming rate. Bacteria may acquire resistance to a given drug by mutation of pre-existing genes or by the acquisition of new genes from other bacteria. One ongoing example of these mechanisms is the evolution of new variants of the TEM and SHV beta-lactamases with altered substrate specificity. PMID- 9746944 TI - Interactions between Mycobacterium tuberculosis and host cells: are mycobacterial sugars the key? AB - Mycobacterium tuberculosis has evolved successful strategies to invade and persist within macrophages. Intimate pathogen-macrophage contacts dictate receptor choice and probably specify the intracellular fate of these microorganisms. Binding to specific receptors, such as complement receptor type 3, could provide an advantage. These interactions appear to involve surface polysaccharides and glycolipids. PMID- 9746945 TI - Causes of death of wild birds of the family Fringillidae in Britain. AB - The provision of supplementary food for wild birds in gardens during the winter months is common in the UK, but it is possible that it may precipitate infectious diseases in the birds. This paper describes the results of postmortem examinations of 116 wild finches carried out over a period of four years. The two commonest causes of death in areas where high mortality had been reported were infections with the bacteria Salmonella typhimurium DT40 and Escherichia coli O86. Coccidia of the genera Atoxoplasma or Isospora were found in several of the birds but were considered to be incidental. Megabacteria were also identified in some of the birds, for the first time in flocks of wild birds in the UK, but they were not considered to be significant. PMID- 9746946 TI - Potential of assisted breeding techniques for the conservation of endangered mammalian species in captivity: a review. AB - An alarming worldwide extinction of animal species is taking place as a result of the activities of the increasing global human population. The original ranges of many animal species are being reduced and fragmented and, in some cases, they have been reduced to perilously small relict populations. The adverse genetic consequences of these restrictions are becoming clear, as are possible methods for their alleviation. The concept of ex situ genetic management of small captive populations of endangered species with a view to re-introducing them into the wild is attracting increasing interest. Modern reproductive techniques will play an important role in such programmes, and it is likely that an increasing number of veterinarians will become involved. However, the literature describing the aims and methods of reproductive genetic management is scattered and often not readily available to interested veterinary surgeons. The aim of this review is to deal with this problem by describing some potential approaches to the captive breeding of endangered species. PMID- 9746947 TI - Inactivation of foot-and-mouth disease virus by heat, formaldehyde, ethylene oxide and gamma radiation. PMID- 9746948 TI - Activity of moxidectin against bots and lungworm in equids. PMID- 9746949 TI - Comparison of a detergent scrub and a swab technique for the quantification of aerobic bacteria on canine skin. PMID- 9746950 TI - Hypertrophic osteopathy associated with ovarian carcinoma in a mare. PMID- 9746951 TI - Calf welfare legislation. PMID- 9746952 TI - Udder oedema in dairy cows. PMID- 9746953 TI - Mastitis in dairy herds associated with Corynebacterium bovis. PMID- 9746954 TI - Study on scrapie resistance. PMID- 9746955 TI - 'Alopecia X' in chows, pomeranians and samoyeds. PMID- 9746956 TI - [Iatrogenic Creutzfeldt-Jakob disease]. PMID- 9746957 TI - [How dangerous is amalgam?]. PMID- 9746958 TI - Salt and hypertension at the close of the millenium. AB - "Can that which is unsavory be eaten without salt?" This question was directed at none other than God by Job, who also had other important problems to ponder. The question posed in this review is the notion that essential hypertension is induced and/or sustained by an unnecessarily high salt intake. If this is indeed the case, then a reduction of salt intake might prevent or effectively treat essential hypertension. A cross-sectional epidemiological study of salt intake in populations showed a positive association of sodium excretion with median blood pressure and the prevalence of hypertension; however, when four disparate populations were deleted, the associations disappeared. A Scottish report on a similarly large population minimized the importance of dietary sodium. A recently analysis of the National Health and Nutrition Examination Survey (NHANES) data base does not support the idea that lower salt intake improves all-cause or cardiovascular mortality; however, the analysis is not without weaknesses. Salt sensitivity is based on the idea that some persons might be more susceptible to salt-induced effects on blood pressure than others. Indeed, several monogenic syndromes exhibit marked salt-sensitivity and their clarification has facilitated our understanding of basic mechanisms. Allelic variants of several genes may be important in salt-sensitive patients with essential hypertension. Meta-analyses of intervention trials in patients with essential hypertension show about a 5 mm Hg decrease in blood pressure with salt restriction. Among the normotensive, this decrease is less than 2 mm Hg. In terms of efficacy, salt restriction has not been shown superior to weight loss or a "vegetable" diet. Nonpharmacological approaches in hypertensive patients should be based on a comprehensive approach. PMID- 9746959 TI - Effects of dental amalgam and its components of histamine release from human basophils and tissue mast cells. AB - Recent studies have shown that metal ions can be released from dental amalgam or other dental materials, and can cause toxic effects on various cells. In this study, the effects of amalgam-conditioned culture medium (ACCM), components of amalgam (Ag+, Cu2+, Sn2+, Hg2+) and dental composite-conditioned culture medium (CCCM) on histamine release from human blood basophils (healthy subjects, n = 3) and tissue mast cells (n = 3) were analyzed. ACCM and CCCM were prepared using either fresh or 6-weeks-aged specimens. Of the metal ions tested, Ag+, and Hg2+ were found to induce histamine release from basophils (Ag+, 0.33 mM: 83 +/- 11% vs Hg2+, 0.33 mM: 100% vs control medium: 5 +/- 5%) and mast cells (Ag+, 0.33 mM: 91 +/- 16% vs Hg2+, 0.33 mM: 99 +/- 1% vs control: 2 +/- 1%), whereas no effects were seen with Cu2+ and Sn2+. Neither ACCM from freshly prepared amalgam nor ACCM from 6-weeks aged amalgam, produced histamine release in basophils or mast cells. Inductively coupled plasma atomic emission spectrometry (ICP) revealed that the Ag(+)- and Hg(2+)-concentrations in ACCM were below the range in which histamine release occurred. Similar to ACCM, no effects on basophils or mast cells were observed with CCCM. In summary, our data show that distinct metal ions present in dental amalgam, can induce (toxic) histamine liberation from basophils and mast cells. However, the amounts of metal ions released from amalgam apparently were too low, to cause histamine release. PMID- 9746960 TI - [Improved kidney function with intravenous prostaglandin E1 in patients with terminal heart failure]. AB - In end stage congestive heart failure activation of a series of compensatory mechanisms increase renal vascular resistance and impair renal function. Prostaglandin E1 is increasingly used in the treatment of severe heart failure for its vasodilating actions. In various experimental settings prostaglandin E analogues are known to improve renal function by modulating renal filtration pressure and redistribution of renal blood flow. However, prostaglandin E1 decreases systemic blood pressure and thus, also renal perfusion pressure, a fact by which renal function might be further compromized in heart failure patients. The aim of the study was to evaluate the effects of prostaglandin E1 on excretory renal function in patients with end stage heart failure and to prove the hypothesis, that the well known local actions of prostaglandins on renal microcirculation might outweigh the negative impact of an expected decrease in perfusion pressure. 25 patients with terminal congestive heart failure were investigated. 13 patients received prostaglandin E1 at a dose of 13.5 +/- 1.9 ng/kg/min in combination with constant rates of dopamine and dobutamine (group A), 12 patients received prostaglandin E1 at a dose of 10.3 +/- 1.7 ng/kg/min without catecholamines (group B). There was no significant difference in prostaglandin dosages between groups. Kidney function was assessed by measuring plasma creatinine and urea nitrogen, urinary output, creatinine clearance, osmotic and free water clearance at baseline and after 72 h of infusion therapy. Hemodynamic parameters were measured by using a balloon tipped pulmonary arterial catheter. Hemodynamic measurements during infusion showed a significant improvement in all patients. At the same time as expected mean arterial pressure decreased in both groups (p < 0.001). Nevertheless, in both groups a significant increase of creatinine clearance during infusion was observed (in group A from 45 ml/min to 78 ml/min., p < 0.05, in group B from 59 ml/min to 105 ml/min., p < 0.001). Creatinine clearance in group B (without catecholamines) reached higher levels than group A (p < 0.05). Urinary volumes did not change during infusion therapy, whereas free water clearance significantly decreased, as an indication of an improvement of renal concentrations ability. We conclude, that in patients with end stage heart failure continuous infusion of prostaglandin E1 improves excretory kidney function. These findings suggest that the local effects of prostaglandin E1 on renal microcirculation can counterregulate the negative impact of prostaglandins on renal perfusion pressure. PMID- 9746961 TI - [Malignant hemangioendothelioma of the thyroid gland: new results on pathogenesis, therapy and prognosis]. AB - BACKGROUND: Malignant hemangioendothelioma of the thyroid is a rare tumor predominantly described in areas with endemic goiter like the Alpine regions. The estimated incidence of the disease is between 0.15 and 0.25 per 100,000 inhabitants per year for Western Austria. The prognosis is reported to be dismal. MATERIALS AND METHODS: Between 1982 and 1995, 10 cases with immunohistochemically confirmed malignant hemangioendotheliomas of the thyroid were referred to our department for postoperative or palliative treatment. Two patients with clear margins at surgery received no adjuvant radiotherapy and were only observed. By surgery, clear margins (R-0 resection) were achieved in 5, microscopic residuals (R-1) were left in 3, and gross residual disease in 1 patient. One patient had an inoperable primary tumor. Postoperative radiotherapy was administered in 6 cases, 4 of them additionally received the radiosensitizer razoxane. Total tumor doses ranged between 58 and 65 Gy. RESULTS: Local tumor control was achieved in 9 of 10 patients; 4 of 10 lived longer than 4 years. The median survival time has not yet been reached and is presently between 7.5 and 21+ months.--Noteworthy is a complete regression of 2 lung metastases in a 72-year-old man by a combination of vindesine, razoxane and radiotherapy. The patient is still in complete remission under a maintenance therapy with vindesine and razoxane since 14 months.--It may also be of interest that 4 of the 10 patients were strongly exposed to vinyl chloride and other polymeric materials during their occupational life. CONCLUSIONS: This small series may indicate that the outcome of this disease may not be uniformly deleterious, and that the resistance to radiotherapy reported in the literature may be questioned.--The data offer new evidence of the occurrence of vinyl chloride-induced angiosarcomas outside the liver, and support observations which have already been published in case reports. PMID- 9746962 TI - Prevalence of self-reported screening mammography and impact on breast cancer mortality in Austria. AB - Mammography for breast cancer screening has been available in Austria free of charge since 1974 and has been explicitly recommended for mass screening since 1980. The type of screening is opportunistic. Evaluation has to rely on population-based data (mortality, stage distribution, screening prevalence). In a representative cross-sectional study (women aged 40-79 years, n = 566, conducted in 1995) 58% reported at least one screening mammography; the lowest prevalence was found in the age group 70-74 years: 39.3%, the highest in the age group 50-54 years: 71.4%. 23.1% reported at least 2 mammograms within an interval of less than 2 years; lowest prevalence 70-74 years: 10.7%, highest 50-54 years: 35.7%. Age-standardized mortality rate has stabilized since 1985. Since 1980 age specific rates have increased significantly in all age groups > 54 years, but a decreasing tendency in most groups has been noted since around 1990. The incidence rates of stage II or worse tumors have increased significantly since 1982, except in the age groups 40-44 and 55-59 years; however, during the past 3 to 6 years the absolute rates of incidence of advanced tumors has decreased in alle age groups. The reduction of advanced cancers at diagnosis, followed by a reduction of mortality rates is plausible related to screening prevalence. More uniform decreasing trends should be expected in the years to come. If not, discontinuation of the current opportunistic form of screening without monitoring and evaluation, financed by public money, has to be discussed. PMID- 9746963 TI - [Sub-pectoral implantation of defibrillators: intraoperative data and long-term outcome]. AB - The present study examined the perioperative mortality and morbidity and lead related complications in patients who had a defibrillator with a transvenous lead system and subpectoral implantation of the generator. Fifty-four out of 57 consecutive patients (95%) received a transvenous lead system. One patient had an acceptable defibrillation threshold with an additional subcutaneous patch whereas no sufficient defibrillation threshold was found in another 2 patients. Two patients died due to congestive heart failure after implantation. Perioperative complications were observed in 4 patients (7%) including pericardial effusion, pocket hematoma, injury of the plexus brachialis and a pneumothorax. None of these complications required surgical intervention. Fifty-five patients were discharged from the hospital. During 27 +/- 10 months none of these patients died. Lead-related complications were observed in 3 patients (5.5%) including microdislocation in two and a outer conductor fracture in one of the lead. CONCLUSION: Technical advancement such as a non-thoracotomy lead system and smaller devices have made the onc-incision approach and subpectoral implantation of the ICD generator clinical routine. Nevertheless complications related to the lead system can occur. Therefore frequent controls of patients with ICD are necessary. PMID- 9746965 TI - "Choose a lesser of two evils" Vinzenz Kern (1760-1829) and the introduction of vaccination in the Slovenian territories of the Habsburg monarchy--on the 200th anniversary of Jenner's discovery of the small-pox vaccination. PMID- 9746964 TI - [Creutzfeldt-Jakob disease in a dura transplant recipient: first observation in Austria]. AB - Dura mater grafts can lead to Creutz-feldt-Jakob disease (CJD) as late complication (dura-CJD). So far 61 dura-CJD cases have been described worldwide. We report here the first dura-CJD case in Austria. A 50-year-old man had a traumatic open frontobasal skull fracture with tearing of dura mater in 1977. Reconstructive surgery used Lyodura (braun Melsungen AG, BRD). Lyodura was derived from pooled cadaveric dura. Ten years after the dural transplantation, the patient developed gait ataxia, paresthesia of both legs, myoclonus and visual disturbance. CT was unremarkable. EEG showed diffuse unspecific changes. The patient died 5 months after onset of disease. Neuropathological examination showed typical histopathology of CJD. Immunocytochemistry detected typical type prion protein (PrP) deposits and scattered PrP plaques in cerebral and cerebellar cortex, basal ganglia and spinal cord. Cerebellar white matter contained numerous PrP miniplaques. This pattern is unusual for sporadic CJD, but is similar to that in CJD after human growth hormone treatment. In our patient and 13/19 earlier described cases with dural graft covering the cerebrum ("central inoculation"), cerebellar disturbance was the initial symptom. Therefore, cerebellar signs are characteristic as initial symptoms in iatrogenic CJD, irrespective of central (cerebral dura mater graft) or peripheral inoculation (e.g., human growth hormone treatment). These data do not support the hypothesis that primary cerebellar symptoms in iatrogenic CJD after peripheral inoculation reflect migration of the infectious agent from the periphery via spinal cord and cerebellum to the cerebrum. PMID- 9746966 TI - ["Auschwitz, NS-medicine and its victims" Comments by 3 human genetics scientists on the book by Ernst Klee]. PMID- 9746967 TI - [Epidemiology of schizophrenic disorders]. AB - Epidemiological research has demonstrated that schizophrenic disorders disorders occur in all countries and in all cultures with a relatively comparable frequency and with the same symptomatics. Furthermore, it was also demonstrated that the incidence of this disorder has not increased in the past 180 years. The clinical epidemiological findings have activated various research approaches: The protective effect of estrogen appears to explain the later occurrence of schizophrenic disorders and their more favourable course in women. Other areas of applied epidemiology (comorbidity research, ecological and social epidemiology) and their effects on suicide prevention are also discussed. Thanks to improved knowledge of prevalence and incidence, of distribution and the various social and ecological factors in schizophrenic disorders epidemiological research has been able to demonstrate that it has acquired increasing significance for basic research. PMID- 9746968 TI - [The course of schizophrenic illnesses]. AB - There is only a limited amount of safe knowledge on the course of schizophrenic psychoses, although 100 years have passed since their first description as "Dementia praecox". 2 main reasons may account for this: On the one hand most studies suffer from more or less serious methodological shortcomings, which do not allow to generalize the results. On the other hand schizophrenic psychoses are possibly not one homogeneous disease with one uniform etiology and a relatively uniform course, but rather a group of diseases with hetereogenous causes and therefore hetereogenous courses as well. PMID- 9746969 TI - [Problems in evaluating new antipsychotic drugs]. AB - A number of novel antipsychotics were registered and introduced into clinical practice in the last decade. These include olanzapine, quetiapine, risperidone, sertindole and zotepine as well as ziprasidone, which is still in the registration process. It quickly became apparent, that it is not always easy to translate results from phase II and III clinical trials into everyday clinical practice. In this context, we discuss methodological aspects that mainly deal with selection of patients for clinical trials and clinical trial methodology. Next to that, an overview of the current knowledge concerning novel antipsychotics is given. There is no doubt that these drugs broaden the therapeutic spectrum made available to patients suffering from schizophrenia. On the other hand, it is evident that there is still a need for a critical evaluation of the risk-benefit-ratio of novel antipsychotics. Clinical psychiatrists also face the challenge to modify some of the traditional treatment approaches. These prerequisites will allow the embedding of novel antipsychotics into modern integrative treatment concepts of schizophrenia. PMID- 9746970 TI - [Rehabilitation in schizophrenia--guidelines for including psychosocial measures]. AB - Today schizophrenic patients live most of their life outside institutions. Social relationships and their quality have, therefore, become practically important. However, they are ambiguous in nature: they can mean a stress for the patient, but they can also be used for therapeutic purposes. It has been shown in many treatment studies, that the combination of psychosocial and medication strategies can improve the long-term outcome of schizophrenia. It is contended that the inclusion of the patient and his or her relatives as partners improves outcome. PMID- 9746971 TI - [Compliance problems in treatment of schizophrenic patients]. AB - Compliance is the degree of adherence to an appropriate medical advise. Especially in longterm treatment, therapeutic success depends largely on patient compliance. In the treatment of schizophrenia with antipsychotic agents a non compliance rate up to 80% is the reason for the difference between the relatively good outcome of controlled treatment studies and the bad results in clinical reality. 4 groups of variables can be identified, which influence compliant behaviour: patient-related factors, factors related to the patients environment, physician-related factors, and medication-related factors. Possibilities to ameliorate the compliance of schizophrenic patients during long-term therapy are discussed. PMID- 9746972 TI - [Schizophrenia--an illness and its treatment reflected in public attitude]. AB - Stigmatization with regard to mental illness and especially schizophrenia has been known from time immemorial. Meanwhile the negative attitudes have become metaphorical for unpredictability, violence, and bizarrely (grotesquely) contradictory behaviour. Persons concerned with these prejudices are excluded from the society and there is little willingness for contact. Particularly afflicted are also the relatives of schizophrenic persons. Media and motion pictures play an essential role in the maintenance of these negative attitudes. There will be suggestions (stimuli), especially by opinion leaders, to reach a change of the attitudes towards diseases and their methods of treatment. PMID- 9746973 TI - [Experimental xenotransplantation]. PMID- 9746974 TI - [Xenotransplantation: what is its future?]. AB - The ultimate solution to the growing shortage of organs for transplantation could be the use of animal organs or tissues. Xenotransplantation of organs of all kinds, size and number would be available to be transplanted to patients with endstage diseases. Advances in technology, as well as the accelerating rate of progress in biochemical, biological and genetic fields, means that it will be possible to bridge times until allografts are available or that successful engraftment becomes possible. Open questions in immunology or metabolism as well as the transfer of infectious material from animal to man need, however, major efforts to be solved. An optimistic outlook can be given, that the problems of xenotransplantation are solved in the next decade, or certainly in the next generation. PMID- 9746975 TI - [Transgenic pigs for xenotransplants for humans]. AB - Microinjection of foreign DNA is currently the only successful procedure to achieve gene transfer in farm animals, though at low efficiency (1-3% transgenic offspring). Integration of the transgene occurs random, expression is normally independent from the number of integrated copies, but can be affected by the site of integration. In most cases, the transgene is inherited according to the Mendelian rules. To achieve xenotransplantation complement regulating gene constructs have been integrated and expressed in transgenic pigs. Upon transfer of hearts from such transgenic pigs to primates the hyperacute rejection response was overcome and an average survival of approximately 40 days was obtained. It is expected that organs from transgenic pigs will be clinically available within the next 5 to 10 years after open questions in particular on the potential transmission of pathogens have been clarified. PMID- 9746976 TI - [Transgenic swine as potential organ donors? Results of the ex-vivo hemoperfusion hDAF transgenic kidney with human blood]. AB - Kidney xenotransplantation is not yet a realistic clinical treatment modality. However, during the last decades more than 30 kidneys from other species have been transplanted into humans; some of the kidneys sustained some function up to 60 days. Recent progress in genetic engineering has raised the possibility to create large transgenic animals which express human complement regulatory proteins (CRP). Since early complement activation is believed to be the main triggering event for xenograft destruction, complement regulation by species specific CRP should avoid hyperacute rejection in transspecies transplantation. The perfusion of hDAF-transgenic pig kidneys with human blood was not associated with the morphological signs of hyperacute rejection when compared to non transgenic control organs. Specific immunohistology could demonstrate that the transgene was sufficient to regulate complement activation beyond C3 despite the endothelial deposition of xenoantibodies. In the future, these organs could be further optimized and ultimately tested in a clinical pilot protocol under appropriate immunosuppression. PMID- 9746977 TI - [Significance of the complement system for xenotransplantation: strategies for therapeutic intervention]. AB - Hyperacute graft rejection triggered by the activation of the recipient's complement system represents the major obstacle to successful xenotransplantation. After the binding of preformed antibodies to vascular glycoproteins complement-induced activation and injury of endothelial cells with subsequent thrombosis leads to rapid destruction of foreign tissues. Inhibition of complement activation is therefore considered as a prerequisite for xenograft survival. Recent animal and cell culture experiments suggest that support of the physiological regulation of the complement system appears to be most promising. Besides the application of soluble complement inhibitors (e.g. soluble complement receptor 1, sCR1; C1 inhibitor) the genetic transfer of human membrane-bound complement regulatory proteins (e.g. DAF, CD59) offers new chances to protect the xenograft against the cytolytic complement attack. Results from the authors' experiments shall be included in a short overview to the issue. PMID- 9746978 TI - [Phenotypic and functional characterization of porcine T-lymphocytes]. AB - Monoclonal antibodies (mAb) against leucocyte differentiation antigens have altered the way in which immunologists examine the immune system. These mAb allow to identify distinct surface molecules on leukocyte populations, by which these cells can be classified, isolated and studied for their functional properties. This review summarises the knowledge about differentiation antigens useful in the characterisation of porcine T-lymphocytes. Furthermore it focuses on several properties of porcine T-lymphocytes and T-lymphocyte subpopulations. PMID- 9746979 TI - [Cellular immune reactivity in xenogenic "human-anti-pig" transplant combination]. AB - The worldwide lack of human organ donors puts the pig as potential xenogeneic donor species into the prime of interest. Aim of the present in vitro study is the analysis of T-cell activation in the clinically attractive combination "pig to-human". Peripheral human blood leukocytes (hPBL) and peripheral porcine blood leukocytes (pPBL) were co-cultured for 4-8 days in the xenogeneic mixed lymphocyte reaction (xMLR) and cell proliferation was measured by 3H-thymidine uptake. Both cell populations were separated into T-cells and antigen presenting cells (APC) to analyze direct and indirect antigen recognition. The results show that (a) activation of human T-cells occurs, (b) the strength of activation depends e.g. on the human responder ("high" and "low" responders), (c) the strength of activation is independent of the responder's HLA-DR status, and (d) direct T-cell activation dominates over indirect activation. Thus, T-cell activation is another immunological barrier that has to be overcome before xenotransplantation can be clinically approached. PMID- 9746980 TI - [Effect of blood viscosity on the function of isolated perfused porcine kidney after cold preservation]. AB - With the new method of gas exchange and dialysis carried out simultaneously with a standard dialysis module, for the first time normothermic ex-vivo whole blood perfusion of pig kidneys becomes practicable under nearly physiological conditions. To show the possibility of animal experiment replacements we use slaughter house kidneys to investigate functional abilities of the perfusion method. Slaughter house kidneys show a state of acute ischemic renal failure due to the unavoidable time of ischemia and cold preservation. Nevertheless they show a specific functional metabolism up to 4 hours of perfusion time. For the future, the new perfusion model seems suitable to investigate acute organ rejection and perfusion phenomena on pig kidneys, which are most interesting for xenotransplantation. PMID- 9746981 TI - [Isolation of islands of Langerhans from human and porcine pancreas for transplantation to humans]. AB - The enormous variability of donor factors and organ procurement related variables prevent a constant isolation success, thus reducing the potential number of clinical islet transplants. Since the availability of intact and viable pancreatic donor tissue intended for islet transplantation is limited, the porcine pancreas was selected as a potential source of xenogeneic islets for human recipients. The differences of islet histomorphology between porcine and human pancreas result in a higher intrinsic fragility of porcine islets during collagenase digestion. Nevertheless, if the isolation method is modified to inhibit factors potentially toxic to pig islets, reproducibility of isolation success is higher in the pig as in the human due to a lower variability in donor characteristics and the opportunity of preselection in regard to age and race. If xenograft rejection can be overcome and the risk of xenosis can be minimized, the logistic prerequisites for xenotransplantation of large amounts of viable pig islets into human recipients with insulin dependent diabetes are fulfilled. PMID- 9746982 TI - [Graft rejection in experimental xenogenic transplantation of isolated islands of Langerhans of the pig]. AB - Prevention of the occurrence of diabetes-specific vascular complications is the final aim of clinical islet transplantation. Pancreatic islets isolated from adult pigs may be a suitable tissue source to transplant a large number of type 1 diabetic patients. Acute cellular rejection may be finally overcome by clinically applicable protocols for tolerance induction. However, primary nonfunction of the graft, as regularly observed in the porcine islet-to-rat xenotransplantation model, may be an additional problem. In this paper, species-specific inflammatory and immunological mechanisms are discussed which prevent early porcine islet graft function in rats but not in mice. PMID- 9746983 TI - [Immuno-isolation of xenogenic islands of Langerhans in a tissue engineered autologous cartilage capsule]. AB - Islet transplantation is a potential cure for diabetes mellitus. The major problem for clinical application remains the prevention of transplant rejection without major side effects. Broad application in early disease will make the usage of xenogeneic tissue necessary. Immunoisolation is an experimental strategy to prevent rejection, by separating the transplanted allogeneic or xenogeneic cells from the host immune system using a barrier device. Current methods of immunoisolation use artificial, not completely inert materials as barrier devices and induce an unwanted foreign body reaction. Using recipient own cells for encapsulation the foreign body reaction could be prevented. This study describes a new method of encapsulation of islets of Langerhans within a capsule of chondrocytes, which may serve as an immunoisolation barrier utilizing the immunoprivileged properties of the chondrocyte matrix and demonstrates the functional survival of the encapsulated islets in vivo. PMID- 9746984 TI - [Occult tumor cells of malignant melanoma and consequences for surgical treatment]. AB - Immediate and complete surgical excision is the standard mode of treatment for primary malignant melanoma. There is still a controversy about the adequate resection margins. In this study we looked for microscopic satellites in order to estimate the extent of local therapy. METHOD: 19 patients with malignant melanoma of the trunk or extremities, treated by wide excision, were included. Clinical courses were documented. Postoperative follow-up was 1.4 years in the mean. In primary tumours and cutaneous excisions we looked for microscopic satellites with routine histology and immunohistochemistry (APAAP-technique, monoclonal antibody HMB-45). RESULTS: Using APAAP-technique in combination with routine histology, we could show that microscopic satellites only occurred in specimens of melanomas of more than 5 mm thickness. In the two cases where evident microscopic satellites were present a systemic dissemination obviously evolved already before the first surgical treatment. Therefore, in those cases wide resection margins do not provide an increasing chance of survival. CONCLUSION: The idea to remove microscopic satellites by wide surgical excisions and thus to reduce the risk for systemic dissemination and local recurrence cannot be supported. PMID- 9746985 TI - [Stepwise concept for treatment of complex anal fistulas]. AB - Most anal fistulas can be easily dealt with by simple fistulotomy. So called complex fistulas-in-ano need a differentiated, individually tailored surgical approach in order to avoid recurrence and sphincter incompetence. Complex fistulas comprise either tracks with high trans-, supra-, or extrasphincteric extension or fistulas that are complicated by multiple side branches, chronic inflammatory disease, previous operations etc. Prior to treatment a thorough preoperative diagnostic work-up is warranted. A precise intraoperative evaluation is paramount to allow radical excision of all inflamed tissue, often necessitating anal sphincter division with subsequent reconstruction. The treatment plan involves staged operations over a period of many months, usually with the (laparoscopic) fashioning of a protective stoma at the primary operation. Analysing our patients in the study period from 1/95 to 12/96 our different surgical approaches and their results are presented and discussed. During this period 96 patients with a fistula-in-ano were operated upon in the Department of Surgery at Wurzburg University Hospital, of which 11 (11.5%) had complex disease. We encountered one early and one late recurrence as well as a parastomal hernia and a stoma prolapse. Anal continence was re-assessed three months following reversal of colostomy. All patients (n = 7) who had perfect continence preoperatively remained unchanged. Preoperatively, four patients were incontinent for gas and liquid stool. Two of these were fully continent, one remained unchanged at re-assessment. The fourth patient did not undergo stoma reversal as yet, because all examinations revealed an incompetent sphincter. This patient is therefore fully incontinent. Successful treatment of complex anal fistulas needs an individual approach and planning over a lengthy period of time, requiring a high level of motivation on the part of both patient and surgeon. PMID- 9746986 TI - [Radical D2,D3 lymph node excision of stomach carcinoma in situs inversus totalis and malrotation of the colon]. AB - The combination of Situs inversus totalis (SIT) with incomplete rotation of the colon is an extremely rare anomaly and especially of interest when presented with advanced gastric cancer. The routine en-bloc resection of gastric cancer with D2,D3 lymphadenectomy according to the recommendations of the Japanese Research Society for Gastric Cancer [5] can, as in this case, present tactical operative problems, due to the mirror-image anatomy and potential vessel anomalies, which might complicate the operative procedure. The preoperative diagnosis of SIT is not only essential to allow an adequate operative approach but furthermore to prevent vessel injury. PMID- 9746987 TI - [Duplication of the stomach as a rare cause of cystic epigastric tumor]. AB - Duplications of the gastrointestinal tract are congenital anomalies seen in about 0.2% of all children. These include the rare gastric duplications. Latter diagnosis is usually made in the first months after birth on recurrent vomiting by detection of an abdominal tumor. The most important imaging modality is ultrasonography. The case of a prematurely born child weighing 1900 g is presented in whom at the age of three weeks a gastric duplication of the greater curvature was diagnosed and who was successfully treated by resection. The postoperative follow-up for 24 months was uncomplicated. PMID- 9746988 TI - [Increased CA-19-9 caused by a splenic cyst: a rare differential diagnosis]. AB - Adenocarcinoma of the gastrointestinal tract, particularly primary adenocarcinoma of the pancreas lead to elevation of serum carbohydrate 19-9 (CA19-9) levels. Some benign disorders, like inflammatory diseases also cause CA19-9 tumor marker elevation. An extremely rare entity are benign splenic cysts which express this tumormarker as reported here. PMID- 9746989 TI - [Laparoscopic versus conventional appendectomy--a comparison with reference to early postoperative complications]. AB - OBJECTIVE AND METHODS: To compare the complications of laparoscopic appendectomy (LA) and conventional appendectomy (CA) 930 consecutive patients from 1989 until 1997 were analysed retrospectively. RESULTS: Conventional appendectomy was performed in 330 patients, laparoscopic in 554 patients and another 46 patients required conversion after laparoscopy. The groups were similar in sex ratio, age and degree of inflammation. Postoperative complications occurred in 8.78%. There were less complications in the LA-group (4.69%) than in the CA-group (13.33%) (p < 0.01), especially wound infections were found less in the LA-group (1.8% vs. 11.21%, p < 0.01). The incidence of intraabdominal abscesses was similar in the LA and CA group (1.44% vs. 1.52%). The differences between the groups are not influenced by complicating appendicitis (perforation or abscess). Systemic complications were similar for LA and CA (0.72% and 0.61%), but were seen more often after conversion (6.52%, p < 0.01). CONCLUSIONS: This retrospective analysis shows that laparoscopic appendectomy significantly reduces postoperative complications, especially wound infections. The authors consider laparoscopic appendectomy to be the procedure of choice in patients with acute appendicitis. PMID- 9746990 TI - [Laparoscopic ligamentum-teres-plasty for treatment of gastroesophageal reflux]. PMID- 9746991 TI - [Anesthesia in surgery]. PMID- 9746992 TI - Benign and malignant prostatic obstruction. PMID- 9746993 TI - Finasteride in the treatment of patients with moderate symptoms of benign prostatic hyperplasia. AB - This was a prospective study involving 27 patients with moderate symptoms of benign prostatic hyperplasia (BPH) treated continuously with 5 mg of finasteride daily for one year. There was improvement in clinical BPH symptoms in 22 patients (81.48%), increase in urinary flow rates by a mean of 2.2 mls/sec in 20 patients (74.07%) and a mean decrease in prostate volume of 20.9% in 25 patients (92.59%) comparable to the findings of the other investigators. No patient on finasteride therapy developed acute urinary retention suggesting reduced risk. The reversal in BPH progression stems from the ability of finasteride to reduce prostate volume thus relieving urinary obstruction and to decrease BPH symptoms and increase urinary flow rates. Finasteride therapy was well tolerated in this study. No adverse effect was observed except impotence in one patient (3.7%) and loss of libido in another patient (3.7%). For symptomatic relief in men with moderate obstructive symptoms of BPH, finasteride should be considered an effective alternative to watchful waiting. These findings warrant further investigations and may signal a positive change in the role of medical therapy in the future long term management of BPH. PMID- 9746994 TI - Screening for urinary tract infection in children with cancer. AB - Neutropaenia and immunosuppression place children on treatment for malignancies at a high risk for infections. We undertook to determine the prevalence of urinary tract infection (UTI) in children on treatment for cancer at the Kenyatta National Teaching and Referral hospital. With the understanding that many laboratories in the rural areas of the country lack appropriate facilities for confirmation of UTI, it was also important to evaluate simple and inexpensive screening methods against a "gold standard" in this cross sectional study. One hundred and eighty six children between the ages of five and 14 years admitted in Kenyatta hospital with leukaemia or lymphoma were enrolled. Besides clinical evaluation, urinalysis and culture and sensitivity were performed on all the subjects. Urine culture was considered the "gold standard" for diagnosis for UTI. The prevalence of UTI was 8.1% (CI = 6.1, 10.1). Only five out of 15 patients were symptomatic. E. coli and klebsiella spp. were responsible for 93.4% of the infections. Presence of pyuria, defined as five or more pus cells per high power field, had a sensitivity of 80.0%, specificity of 97.1% and a positive predictive value of 70.6% while comparative values associated with a positive nitrite test were 60%, 97.7% and 96%. Other clinical and laboratory tests had low sensitivity. UTI is a relatively frequent infection in children on cancer treatment. Screening for pyuria is simple, inexpensive and an accurate method of diagnosing UTI in children on treatment for lymphohaematopoietic malignancies in situations where facilities for urine culture are unavailable. PMID- 9746995 TI - Breast cancer and risk factors in an African population: a case referent study. AB - Numerous studies have investigated different aspects of breast cancer including the known risk factors in the Western world. In sub-Saharan Africa breast cancer in women despite being a public health problem has not received attention from national health providers. This case-referent study investigated the association between a number of risk factors and breast cancer in Tanzania. There was a strong evidence of breast cancer risk increasing with parity chi 2 = 10.6; p = 0.001. However lactation did not show a significant protective effect against breast cancer (OR = 0.6, p = 0.5). Other factors investigated included age at menarche, age at first sex, marital status, age at menopause and duration of menstruation. Statistical analysis revealed that these factors were not significantly associated with breast cancer in these Tanzanian women. The influence of these results are discussed in terms of possible intervention strategies. PMID- 9746996 TI - T-cell subset counts and serum immunoglobulin concentrations in patients with chronic renal failure at the Kenyatta National Hospital. AB - This study was designed to determine whether there was any difference in the T cell subset counts and serum immunoglobulin concentrations in patients with chronic renal failure as compared to normal controls. Ninety individuals participated in the study. These were divided into three groups as follows; (i) 30 subjects with normal renal function; (ii) 30 subjects with chronic renal failure (CRF)(creatinine clearance 10-50 mls/min), not requiring haemodialysis and; (iii) 30 subjects with end stage renal disease (creatinine clearance < 10 mls/min) on haemodialysis. The subjects in the three groups were matched for age and sex. In addition, it was ascertained that none of the subjects was on any medication or suffered from any ailment known to interfere with the immune system. The T-cell subset counts were carried out using flow cytometry while the serum concentration of immunoglobulins was measured using the radio immunodiffusion method. Patients with CRF, whether on haemodialysis or not, had significantly lower lymphocyte counts as a proportion of total white cell count (19% and 19.2% respectively versus 39%) and low absolute CD4 cell counts per mm3 (337 +/- 94 and 449 +/- 116 respectively versus 891 +/- 360) and CD8 cell counts per mm3 (437 +/- 234 and 490 +/- 176 respectively versus 644 +/- 228) as compared to normals, with no statistically significant difference between the two groups with CRF. The CD4: CD8 ratios in the three groups studied were 1.487 +/- 0.233, 0.961 +/- 0.326 and 0.751 +/- 0.167 respectively, being significantly higher in normal controls than in any of the groups with CRF (p < 0.05) and in the group with CRF not requiring dialysis than in those requiring it (p < 0.05). The serum concentration of immunoglobulins in the two groups with CRF were similar to those in the group with normal renal function. It is concluded that CRF represents a state of immunodeficiency not significantly corrected by haemodialysis. PMID- 9746997 TI - Minimal invasive surgery: a district hospital experience. AB - This was a prospective analysis of the first 162 patients who underwent biliary and nonbiliary minimally invasive (video laparoscopic) procedures in the Royal Commission Medical Centre (RCMC) over two periods separated by a one year interval (September 1993-September 1994)-(October 1995-February 1996). One hundred and fifty patients had video laparoscopic cholecystectomy (VLC). Thirty four males and 116 females with a mean age of 39.7 years (range 16-80). Forty two patients (28%) were admitted as emergency (37 acute cholecystitis, 5 acute pancreatitis). The indication for VLC was symptomatic gall stones. The VLC was accomplished successfully in 144 patients (96%). Six patients (2 electives and 4 emergency) required a conversion for various reasons, unfavourable anatomy being the commonest. Ten patients with preoperative evidence of a dilated common bile duct, with or without stones had an ERCP done in another hospital 200 km away. The median operative time was 100 minutes (range 30-270 minutes) There were three major complications (one CBD injury, one bleeding from gall bladder bed and one post operative acute pancreatitis) and 6 minor complications (urethral bleeding, atelectasis post-operative pyrexia, umbilical port cellulitis, prolonged ileus and acute anxiety state). The median hospital stay was 72 hours for successful VLC. Twenty five per cent of the patients did not require any narcotic analgesic. Twelve patients (7.4%) had one or another non-biliary video laparoscopic procedure. Our results suggest that VLC can be offered and performed safely in the majority of patients presenting with acute and/or chronic cholecystitis and that the results we achieved in a district hospital are comparable to other series. We conclude that VLC will continue to be demanded by patients and non biliary video laparoscopic procedures which were slow to develop in our hospital will continue to need special training, interest and expertise before it can be adopted as a routine. PMID- 9746998 TI - Acute rheumatic fever in southern Saudi Arabia. AB - Forty six attacks of acute rheumatic fever (ARF) in forty patients were diagnosed between November 1987 and August 1995. Thirty four were initial attacks and 12 were recurrences. Arthritis was the commonest feature, 84.8%. Carditis occurred in 65.2% of the group, 67.6% of the initial attacks and 58.3% of the recurrences; however, the frequency of moderate/severe carditis was higher in recurrences, 25% versus 11.8%. Of those with carditis, mitral regurgitation occurred in 93.3%, aortic regurgitation in 16.7% and significant tricuspid regurgitation in 6.7%. Mitral stenosis was not encountered. No mortality occurred during ARF. Chorea, erythema marginatum and subcutaneous nodules were infrequent. These data are similar with those from a previous study which demonstrated the mild nature of ARF in Saudi Arabia, but showed higher frequency of carditis and suggested the frequency of carditis was not significantly higher during recurrences as compared to frequency of moderate/severe carditis. PMID- 9746999 TI - Arrival in the labour ward in second stage of labour--any prognostic significance? AB - A comparative descriptive study was carried out to determine whether, in uncomplicated term pregnancies with the foetus in vertex presentation, there were any differences in maternal or foetal outcome between women who arrived in the labour ward in second stage of labour and those who arrived in early active phase. There were two hundred and seventeen women each in the study and comparison groups. There were no significant differences between the two groups as regards age, parity, marital status and level of education. Women in the comparison group were better antenatal clinic attendants. Those in the study group were more likely to have indicated that they had problems with transportation. They also had considerably shorter labours and all achieved spontaneous vaginal deliveries; a significant proportion (10.6%) of the comparison group had interventional deliveries. The incidence of episiotomies, lower genital tract injuries, manual removal of placenta and postpartum haemorrhage after vaginal delivery were not different between the two groups. Babies born to mothers in the study group were significantly lighter, by about 170 gms, and had a lower incidence of low one-minute Apgar scores. There were no significant differences in the rates of admission to the neonatal intensive care unit or in early neonatal deaths. Arrival in the labour ward in second stage of labour prognosticates non-interventional delivery without any increased risk of adverse outcome to the mother or her baby. PMID- 9747000 TI - Prevalence of intestinal parasites among children in southern Sudan. AB - A study was conducted in southern Sudan to determine the prevalence of intestinal parasites among school children. A total of 275 stool samples which were examined using formol-ether concentration techniques yielded 15 different species of parasites. Hook worm with a prevalence of 13.1% was the predominant nematode followed by Strongyloides stercoralis (3.3%), Trichostrongylus (2.5%), Schistosoma mansoni (2.2%) and Trichuris trichiura (1.8%). Ascaris lumbricoides and cestodes were not detected in this population. Intestinal protozoans were common. Entamoeba coli (37.8%), Entamoeba histolytica (28.4%) and Giardia lamblia (9.8%). Children in the age group 6-10 years old were the most affected followed by the 11-15 year-old age group. The infection rate was slightly higher in males than females. PMID- 9747001 TI - Effect of insecticides on vital activity, hepatic enzymes and red blood cell acetyl cholinesterase activity of rabbits in Makkah. AB - Developing animals and invertebrate are markedly more sensitive to acute toxicity through exposure to insecticides. Varieties of insecticides are used for hygienic control in Makkah holy places. The present study examines the acute effects of commonly used insecticides in Makkah area. Rosfin as an organophosphorus, Airlen as a pyrethroid and Sulvac as a carbamate derivative were tested for their effects on vital activities, hepatic transaminases, serum triglycerides and acetyl cholinesterase activity of rabbits. The insecticides were tested in same and double concentration used for insect control by Makkah's municipal authorities. Compressed air was used as a source of pressure for spraying wooden boxes designed for habitation of animals during the experiments. There were no significant changes in vital activities of rabbits in both concentrations. However, serum glutamate pyuvate transaminase (SGPT) and serum glutamate oxaloacetate transaminase (SGOT) showed irregular changes (mild decrease or increase) in all groups, while triglyceride showed mild rise after six days exposure in case of double concentration. Acetyl cholinesterase showed mild increase in activity after five minutes incubation time, but there was unnoticed increase in activity after 15, 25, 35 and 45 minutes of incubation. In case of Airlen, the activity increased after five minutes of incubation and decreased thereafter. In conclusion, insecticides used in the holy places of Makkah area have no apparent effects on vital activity, acetyl cholinesterase activity and showed no significant effect on rabbit hepatic transaminases and serum triglyceride. PMID- 9747002 TI - Predictors of low birthweight at the community level. AB - The outcome of pregnancy was studied in 148 women over a two year period in a rural area of Kenya as part of a prospective longitudinal study whose main objective was to study the functional effects of mild to moderate malnutrition. Data were collected on maternal anthropometric variables monthly, haemoglobin levels were determined by blood samples taken every six months, food intake was based on two days each month of actual weight and recall. Each woman's past reproductive history was established at the beginning of the study. Birth weight was taken and recorded within seventy two hours of delivery. Discriminant analysis was used to identify predictors of low birthweight. The analysis was based on 123 cases who had complete data on all the variables used in the equation. Of those included in the analysis, 14 women (11%) delivered low birthweight babies and 109 had normal birthweight babies. Results of the discriminant analysis showed that mid upper arm circumference (MUAC), body mass index (BMI), Blood haemoglobin levels (HB) and socioeconomic status (SES), are the best predictors of low birthweight. Ranked in order of relative contribution to birthweight they are BMI, HB, MUAC and SES. Low birthweight prevalence was determined as being 11.2 per cent. Eighty per cent of all known cases were correctly classified using the four variables. As a screening tool for low birthweight this model with four variables has 93% sensitivity, 78.4% specificity, 35.13% positive predictive value and 98.98% negative predictive value. The results suggest that it is possible to identify women at high risk for delivering low birthweight babies at the community level. PMID- 9747003 TI - Hypertension in east Africans and others of African descent: a review. AB - As the number of fatalities from cardiovascular diseases declines in western industrial nations, an opposite trend is observed in the East African region. Inter-regional variations in the prevalence of vascular disorders have been attributed to socioeconomic, psychosocial and heritable physiological parameters. Although faulty mineralocorticoid metabolism and the dysfunctional kidney are prominent features of circulatory problems, many current studies are focused on membrane receptors, transmembrane ion transport mechanisms, ion channels and the possible genetic polymorphisms that determine the characteristics of those molecular structures in the vascular system of normal or hypertensive persons. In this review, a composite of the data available on each of the above parameters and its significance in the pathogenesis of hypertension in the industrial West and transforming economies of East Africa is presented. PMID- 9747004 TI - Facial and ear dimensions in term Nigerian neonates. AB - A study of selected facial and ear dimensions in 200 term and appropriate-for gestational age neonates delivered at the University College Hospital, Ibadan, Nigeria, was carried out with the aim of describing their normal range of variation and providing reference values for clinical use. The features studied included inner canthal distance, outer canthal distance, palpebral fissure length, nasolabial distance, oral intercommissural length, total ear length and ear length above the eyeline. The inner canthal distance ranged from 1.6 to 2.5 cm with a mean of 2.1 (SD 0.2) cm while the outer canthal distance ranged from 5.2 to 7.2 cm with a mean of 6.1 (SD 0.4) cm. The oral intercommissural length ranged from 2.5 to 4.0 cm with a mean of 3.1 (0.3) cm and the total ear length ranged from 2.4 to 4.0 cm with a mean of 3.2 (SD 0.3) cm. Three per cent of the neonates had the whole ear located completely below the eye line. No significant sex differences in the mean values of any of the dimensions studied were found. The findings of the study are consistent with those of other workers who have documented shorter ears in the Negro neonate compared to his Caucasian counterpart. It is suggested that local values derived from well-defined populations be used as reference in the evaluation of the child with dysmorphic features in order to avoid errors of classification due to racial variations in the range of normal. PMID- 9747005 TI - Umbilical cord prolapse: a clinical study of 60 cases seen at Obafemi Awolowo University Teaching Hospital, Ile-Ife. AB - The incidence of umbilical cord prolapse at Obafemi Awolowo University teaching hospital complex, Ile-Ife over a ten year period was 0.42% (one in 240 deliveries). The incidence was observed to be higher among the unbooked patients (76.7%). Analysis of the 60 cases reveals that multiparity, unengaged presenting part from cephalo-pelvic disproportion, prematurity, prelabour spontaneous rupture of membranes, breech presentation, and multiple pregnancy were the major contributory factors. The perinatal mortality (36.7%) was significantly higher than that of the hospital which was 8% (P < 0.05). The perinatal mortality rate was higher among the unbooked patients (86.4%). Caesarean section gave better results except when the cervix was fully dilated. Early resort to Caesarean section, proper and adequate antenatal care and properly supervised hospital delivery is recommended. PMID- 9747006 TI - Investigation into the cercariacidal and miracidiacidal properties of Endod (Phytolacca dodecandra) berries (type 44). AB - Aqueous extract of ground Endod (Phytolacca dodecandra) berries (Type 44) was investigated for its cercariacidal and miracidiacidal properties. Aqueous extract of the berries prevented snails from being infected by miracidia at a concentration of 4 ppm. Assessment of cercariacidal activity of Endod berries indicated that mortality of cercariae exposed to aqueous extract of Endod berries increased with increase in concentration of the test material and exposure time. Viability assessment test showed that pre-treatment of the cercariae with 12 ppm of the extract completely inhibited infection of mice by cercariae and significantly reduced tissue egg deposition and worm establishment in the mice (ANOVA, P < 0.05) The potential use of (Phytolacca dodecandra) berries against schistosome larval stages in fresh water in a schistosomiasis control program is discussed. PMID- 9747007 TI - Re: Sexually transmitted infection: agents to look out for. PMID- 9747008 TI - [Inflammatory intestinal diseases and their extra-intestinal manifestations observed at Liban]. AB - 101 cases classified as inflammatory bowel disease at the Hotel-Dieu de France Hospital between 1982 and 1994 were investigated. Files containing a sure diagnosis and complete clinical and biological investigations were included. 65 files were retained for the study: 48 cases had ulcerative colitis (UC), 17 had Crohn's disease (CD). UC averaged 4.7 admissions per 10,000 admissions to the hospital, while CD averaged 1.54 per 10,000. The UC/CD ratio was 2.8 and the Female/Male ratio was 1.4 for UC and 0.9 for CD. Medium follow-up was 6.4 years. 30% of our group was followed for more than 10 years. Overall frequency of extra intestinal manifestations was similar to that reported worldwide. This is especially true for UC. However, no cases of ankylosing spondylitis, sclerosing cholangitis or erythema nodosum were found. One case of pyoderma gangrenosum was found in the series of CD. Extra-intestinal manifestations were found in 54% of UC patients and 94% of CD patients, probably because milder cases of this disease were misclassified as infectious diarrhea. PMID- 9747009 TI - Earthquakes: health outcomes and implications in Lebanon. AB - Earthquakes remain damaging to human life and property. Recent reports point out that there is 50% chance to have a damaging earthquake in the next 50 years in Lebanon. Major health outcomes of earthquakes include physical injuries, cardiovascular effects and psychological reactions. Physical injuries are correlated with entrapment, higher number of floors in buildings and behavior at time of impact. Cardiovascular effects of earthquakes are either immediate or delayed. Immediate cardiovascular effects are exemplified by increased coronary death while delayed effects are reflected by the increase in coronary risk factors such as serum triglycerides and heart rate. The severity of PTSD decreases as the distance from the epicenter of the earthquake increases. Preventive measures such as implementation of safety building codes, medical emergency readiness and public education should be carried out in areas threatened by earthquakes. Also special mental health programs should be executed following an earthquake. PMID- 9747010 TI - Management of posttraumatic posterior urethral disruption. AB - We reviewed our experience with 17 cases of posterior urethral disruption due to traumatic pelvic injuries. In all cases, a suprapubic cystostomy was performed at first. For blunt injuries, urethroplasty was delayed for 6 months in average. For most of the penetrating injuries (3/4), we performed immediate debridement and primary repair. Resulting bulbous or membranous strictures less than 3 cm long were treated with one-stage perineal excision-reanastomosis urethroplasty. Membranous strictures longer than 3 cm were managed with a combined transpubic perineal repair, while bulbous defects longer than 3 cm were treated with a scrotal pedicled island flap. The overall restricture rate was 25%. Those having had initial repeated urethrotomies displayed a 100% restenosis rate. Incontinence rate was 12.5% Erectile dysfunction occurring in 42% of our patients is a sequela of the pelvic injury and was found to be directly related postoperatively to its presence at the time of surgery. PMID- 9747011 TI - Breast cancer screening: recommendations and controversies. PMID- 9747012 TI - Cardiac arrhythmias and the autonomic nervous system. PMID- 9747013 TI - [Auricular vulnerability: its definition and practical application]. PMID- 9747015 TI - Long-term management of atrial fibrillation. PMID- 9747014 TI - Antiarrhythmic drugs: how to use and to evaluate them. PMID- 9747016 TI - [Test of inclination or tilt test: concerning children and adolescents]. PMID- 9747017 TI - [Management of Wolff-Parkinson-White syndrome]. PMID- 9747018 TI - [Profile of the care of arterial hypertension: questionnaire addressed to Lebanese physicians]. PMID- 9747019 TI - Drug related illness leading to hospitalization at the American University of Beirut-Medical Center. PMID- 9747020 TI - The role of baccalaureate nursing students in the matrix of health promotion. AB - The Declaration of Alma Ata in 1978 put primary health care into the forefront of government health agendas around the world. The Ottawa Charter followed in 1986, emphasizing the role of health promotion for enhancing the wellness of populations and the potential cost savings related to illness care. The findings from research and evaluation in health promotion indicate that multiple approaches, such as mass media campaigns, school-based education, community education, community development, environmental initiatives, and implementation of legislation, are most likely to bring about sustained change (Garrard, 1990). Health education is a component of health promotion and is directed toward enhancing the skills of individuals, families, and communities to change behavior. Behavior change, however, will not be achieved by isolated health education campaigns but by a combination of strategies, methods, and activities at a local, state, and federal level. The type of campaigns that are implemented by nursing students is one intervention that forms part of the matrix of health promotion. This article describes the experience of a School of Nursing in the application of health education by students in a variety of settings in the community. These campaigns help the students to connect theory to nursing practice in a creative way that provides immediate and long-term benefits and may enhance the health of the community. PMID- 9747021 TI - A comparison of traditional folk healing concepts with contemporary healing concepts. AB - Health professional interest in folk healing concepts as a means of complementing contemporary health care has flourished within recent years. Yet, despite this seemingly new area of study, many basic tenets of folk healing are consistent with principles ascribed to by modern health professionals. In this article, I elucidate several concepts shared by traditional folk healing systems and the contemporary health care system. These concepts include origin of illness, harmony and balance, motion, colors, symbols, and family and community involvement. The analysis of these concepts should enable nurses to have an advanced understanding of transcultural healing practices, as well as a heightened perspective on the shared dimensions of folk and contemporary health care systems. PMID- 9747022 TI - Eastern Indians' childbearing practices and nursing implications. AB - The purpose of this article is to assist nurses to understand and provide culturally sensitive perinatal care to Eastern Indians residing in the United States. Traditional practices during pregnancy, labor and delivery, and after delivery are described. The strategies that are culturally relevant and sensitive to the needs of the clients are suggested. PMID- 9747023 TI - Breastfeeding among low-income women with and without peer support. AB - This research examined the effect of peer support on breastfeeding duration and exclusivity (breastfeeding without supplements) in a population of low-income women during the first 3 months postpartum. Participants in the peer counselor group (n = 18) exhibited higher rates of exclusive breastfeeding across time than those without a counselor (n = 18), and more exclusive breastfeeding was associated with long duration overall. Mother's career plans had the greatest effect on duration of breastfeeding. Women who intended to return to work, attend school, or both breastfed 6 to 9 weeks less than participants who intended to stay home. Attendance at a breastfeeding class and knowing someone who had breastfed was significantly correlated with a longer duration of breastfeeding. Nutritionists from the Women, Infants and Children (WIC) Program were the primary source of breastfeeding information. Two main factors discouraged women from breastfeeding: returning to work, school, or both and the perception of a diminished milk supply. Greater emphasis should be placed on prenatal breastfeeding education for low-income women, and their mothers and grandmothers should be included. Peer support is one important component of social support in the area of breastfeeding that community health nurses (CHNs) can utilize. CHNs are in a unique position to assist working mothers, provide support, and develop educational programs to enhance breastfeeding success in this population. PMID- 9747025 TI - VNTR (variable number of tandem repeat) sequences as transcriptional, translational, or functional regulators. AB - VNTR (variable number of tandem repeat) markers, also called single-copy minisatellites, were originally isolated from human DNA as highly informative restriction fragment length polymorphisms for mapping purposes. Evidence has lately emerged that some VNTR sequences play significant roles in the regulation of transcription, and that some may also influence the translational efficiency or stability of mRNA, or modify the activity of proteins by altering their structure. Some apparent associations of VNTR sequences with personality traits or with susceptibility to diseases have strengthened the likelihood that these tandemly-repeated genomic elements are of physiological and biological importance. In this review, we summarize recent progress in efforts to clarify mechanisms involving VNTR sequences. PMID- 9747024 TI - Preschool children at risk for repeat injuries. AB - Home-related injuries are a major problem in young children. This study investigated maternal and child characteristics related to repeat injuries in preschool children. A secondary data analysis was conducted on a national probability sample of African American and White mothers who participated in the longitudinal study of the National Maternal and Infant Health Survey. Repeat injuries in the African group were associated with poor maternal health status, maternal use of alcohol, and Ipecac in the home. Significant factors for the White group were male child, unmarried mother, difficulty managing the child, maternal depression, poor maternal health status, and Ipecac in the home. Findings support the need to identify high-risk children and to provide interventions specially aimed at amelioration of those factors related to repeat injuries. PMID- 9747026 TI - Association and linkage of LDLR gene variation with variation in plasma low density lipoprotein cholesterol. AB - The role of common variation in the low density lipoprotein (LDL) receptor gene (LDLR) as a determinant of variation in plasma LDL cholesterol in normolipidemic populations is not well established. To address this question, we used both association and linkage analysis to evaluate the relationship between plasma LDL cholesterol and genetic variation in LDLR and in three other candidate genes for lipoprotein metabolism, namely, APOE, PONI, and LPL. We studied a sample of 719 normolipidemic Alberta Hutterites, who comprised 1217 sib pairs. Variation in each of the four candidate genes was significantly associated with variation in plasma LDL cholesterol, but the average effects of the alleles were small. In contrast, sib pair analysis showed that only the LDLR gene variation was linked with variation in plasma LDL cholesterol (P = 0.026). Thus, the common LDLR gene variation was both associated with and linked to variation in plasma LDL cholesterol, suggesting that there is a functional impact of structural variation in LDLR on plasma LDL cholesterol in this study sample. However, the absence of linkage of variation in LDL cholesterol with the other three candidate genes, in particular APOE, is consistent with a lower sensitivity of linkage analysis compared with association analysis for detecting modest effects on quantitative traits. Attributes such as the genetic structure of the study sample, the amount of variance attributable to the locus, and the information content of the marker appear to affect the ability to detect genotype-phenotype relationships using linkage analysis. PMID- 9747027 TI - A novel missense mutation of the tissue-nonspecific alkaline phosphatase gene detected in a patient with hypophosphatasia. AB - Hypophosphatasia is a rare heritable inborn error of metabolism characterized by abnormal bone mineralization associated with a deficiency of alkaline phosphatase. The clinical expression of hypophosphatasia is highly variable, ranging from death in utero to pathologic fractures first presenting in adulthood. We investigated the tissue-nonspecific alkaline phosphatase (TNSALP) gene from a Japanese female patient with hypophosphatasia. By a quantitative polymerase chain reaction (PCR) method, the amount of TNSALP mRNA appeared to be almost equal to that in normal individuals. Gene analysis clarified that the hypophosphatasia originated from a missense mutation and a nucleotide deletion. The missense mutation, a C--> T transition at position 1041 of cDNA, results in an amino acid change from Leu to Phe at codon 272, which has not yet been reported. The previously reported deletion of T at 1735 causes a frame shift mutation downstream from Leu at codon 503. Family analysis showed that the mutation 1041T and the deletion 1735T had been inherited from the proband's father and mother, respectively. An expression experiment revealed that the mutation 1041T halved the expression of alkaline phosphatase activity. Using homology analysis, the Leu-272 was confirmed to be highly conserved in other mammals. PMID- 9747028 TI - Heterozygosities and allelic frequencies of 358 dinucleotide-repeat marker loci in the Japanese population. AB - We examined 64 normal Japanese chromosomes to determine the heterozygosities and allelic frequencies of 358 dinucleotide-repeat marker loci spanning the whole human genome. Comparisons of the data for each marker in the Japanese population sample with data for the same markers among Caucasian samples in the Genome Database (GDB) revealed a slightly lower average of heterozygosity in Japanese (71% vs 79%). Although the majority of the markers were as informative as in Caucasians, some in our sample were uninformative due to low heterozygosity; 38 loci revealed heterozygosities lower than 50% and 11 of these were less than 30%. Furthermore, allelic distributions at many of the marker loci were quite different in the two racial groups. Since such differences will influence statistical analyses between markers and disease loci, our data will be essential for linkage analyses, sib-ship pair analyses, and association studies involving the Japanese population. Therefore we have archived this database on a home page on the Internet (http:@www.ims.u-tokyo.ac.jp/nakamura/Yamane.html++ +). PMID- 9747029 TI - Expression and chromosomal localization of KIAA0369, a putative kinase structurally related to Doublecortin. AB - Neuropathy in vertebrates can be a consequence of failure of genes involved in the nervous system to be expressed at the correct times and levels during embryonic life. Recently, a brain specific gene, Doublecortin, was cloned and was shown to have mutations in X-linked lissencephaly and double cortex syndrome. KIAA0369 is a putative kinase that is structurally related to Doublecortin. We compared the expression of KIAA0369 with that of Doublecortin, both of which were expressed specifically or predominantly in fetal brain among 20 different tissues examined. The deduced products of both genes contain a unique domain (the Doublecortin [DC] domain), but KIAA0369 also contains a calmodulin-dependent kinase (CaM kinase)-like domain following the DC domain. We found at least four splicing variants of KIAA0369: KIAA0369-AS (type A, short version), KIAA0369-AL (type A, long version), KIAA0369-BS (type B, short version), and KIAA0369-BL (type B, long version). KIAA0369-B, which lacked the DC domain and maintained the kinase domain, was expressed in adult as well as fetal brain, but the variants that included the DC domain, KIAA0369-A, were expressed predominantly in fetal brain. These results suggest that the DC domain plays an important role in the development of the nervous system. In the adult brain, KIAA0369 was expressed in all 15 different regions examined, more intensely in cerebral cortex, occipital pole, frontal lobe, amygdala, and hippocampus, and less intensely in corpus callosum and thalamus. The murine homologs of Doublecortin and KIAA0369 were not detectable in 7-day mouse embryos, but both genes were expressed extensively in 11-day embryos. Human KIAA0369 was mapped by fluorescence in situ hybridization (FISH) to chromosome 13q13-q14.1. The presence of genes related to neuropathy has been reported in this locus. PMID- 9747030 TI - Interstitial deletion of chromosome 7q in a patient with Williams syndrome and infantile spasms. AB - Interstitial deletion of 7q11.23-q21.11 was identified by cytogenetic methods in a 4-year-old boy with Williams syndrome (WS) and infantile spasms. Deletion of the elastin (ELN) gene and the DNA polymorphic markers, D7S1870, D7S2490, D7S2518, and D7S2421, were identified in the patient, but the loci for D7S653 and D7S675 were not involved. Zackowski et al. (1990) reported that 6 of 16 patients with the interstitial deletion of 7q11.2-q22 had abnormal electro encephalograms, or seizures, or both, and that infantile spasms were present in 2 of the 6 patients. WS is a well defined developmental disorder characterized by distinct facial features, gregarious personality, and congenital heart defects. Seizures are not generally associated with this syndrome. WS commonly is characterized by deletion of the loci for ELN and D7S1870, but not those for D7S2490, D7S2518, or D7S2421. This suggests that a gene responsible for infantile spasms is located in the 2.7-cM interval between loci D7S1870 and D7S675. PMID- 9747032 TI - Molecular phylogenetics of the hominoid Y chromosome. AB - The Human Y-chromosome plays a central role in sex determination, and is composed of DNA sequences homologous to the Y-chromosome, families of Y-specific repetitive DNA sequences, and single copy sequences. We investigated the chromosomal location of Y-specific DNA sequences, in the chimpanzee (Pan troglodytes), gorilla (Gorilla gorilla), and orangutan (Pongo pygmaeus) by the fluorescence in situ hybridization (FISH) technique. The Yq subtelomeric DNA sequences (DYS427) have been observed to be intact at the presumed loci. Also, the amelogenin gene (AMELY, Yp11.2) revealed sequence homology and positional conservation in the higher primates, except in gorilla where positional divergence was observed. PMID- 9747031 TI - A novel mutation of coproporphyrinogen oxidase (CPO) gene in a Japanese family. AB - Hereditary coproporphyria (HCP) is an autosomal dominant disease characterized by a deficiency of coproporphyrinogen oxidase (CPO). Only 11 mutations of the gene have been reported to date as the mutations responsible for HCP. We report here a novel mutation of the gene responsible for the disease in a Japanese family. Analysis of the polymerase chain reaction (PCR) amplified DNA fragments of the gene by direct-sequencing and/or cloning-based sequencing methods revealed the gene abnormality responsible for the disease. The mutation found was a single base deletion of T at nt position 526, which results in frame shift and truncation of coded protein at amino acid position 204. Screening of pre symptomatic cases seemed to be possible by PCR restriction analysis using restriction enzyme Xcm I. PMID- 9747033 TI - Identification of amplified DNA sequences on double minute chromosomes in a leukemic cell line KY821 by means of spectral karyotyping and comparative genomic hybridization. AB - Double minute chromosomes (dmin) are cytogenetic hallmarks of amplified genes. Using spectral karyotyping (SKY) and comparative genomic hybridization (CGH), we identified the origin of amplified DNA in a leukemic cell line, KY821, that harbors numerous dmin. The SKY revealed that the DNA sequences of dmin are derived from materials of chromosome 8, and CGH showed a high degree of overrepresentation only at 8q22-24, indicating that in KY821 only chromosomal material of 8q22-24, containing MYC, is amplified in dmin. An approach combining SKY with CGH should facilitate efforts to identify novel chromosomal regions of gene amplification and contribute information about genetic lesions that underly neoplastic tumors. PMID- 9747034 TI - Joint laxity, vitreoretinal degeneration, facial abnormalities, and generalized skeletal alterations: a new syndrome? AB - A Japanese girl with a hitherto unknown combination of malformations is reported. The cardinal features included hyperextensibility of the joints, vitreoretinal degeneration with cataracts, and facial abnormalities, comprising hypertelorism, prominent eyes, downslanting of the palpebral fissures, mid-face recession with a short nose, deformed auricles, and microretrognathia with a high arched palate. Skeletal survey revealed multiple wormian bones, hypoplastic facial bones and mandible, narrow thorax with wavy ribs, narrow ilia, and coxa valga with slight broadening of the proximal femora, findings of which were individually minor, but the assemblage of which assisted in the syndromic identification. Although skin biopsy did not contribute to the causal clarification, it was tempting to speculate that the syndromic constellation of the present disorder resulted from an underlying defect of connective tissues. PMID- 9747035 TI - Y-specific DNA polymorphisms of the YAP element and the locus DYS19 in the Korean population. AB - The Y Alu polymorphic (YAP) element (DYS287) and the Y-linked tetranucleotide microsatellite locus DYS19 were examined in samples from a total of 455 unrelated males in the Korean population. The frequency of the YAP allele was found to be 1.3% (6/455) in the Korean population. These results are consistent with previous reports that showed the YAP element to be absent in most Asian populations, with the exception of the high frequency of the YAP allele in the Japanese population. All five common alleles at the DYS19 locus were identified in this study, The C allele was the most frequent (197/455), followed by the D (119/455), B (78/455), E (41/455), and A (20/455) alleles. Seven combination haplotypes (DYS287/DYS19) were found, and the mean combination haplotype diversity in the Korean population appeared to be 0.71. Based on results of these two loci, Japanese and Korean populations may share some common genetic structure that could reflect recent gene flow and some amount of admixture of Y chromosomes between these two populations. PMID- 9747036 TI - Novel MEN1 gene mutations in familial multiple endocrine neoplasia type 1. AB - The recent isolation of the gene responsible for multiple endocrine neoplasia type 1 (MEN 1) has enabled direct genetic diagnosis for people with endocrine tumors and family members of affected patients. Although MEN 1 is rarely recognized in the Japanese population compared to its prevalence in Caucasians, we have previously reported a high prevalence of this disease in a limited area (Nagano Prefecture; population, 2.15 million). In this communication, we report mutations of the MEN1 gene in kindreds living in Nagano Prefecture. The absence of a common mutation among these kindreds indicates that the high prevalence of MEN 1 in this area is not due to a regional accumulation of patients descended from a common ancestor. This result implies that the prevalence of MEN 1 in other areas of Japan could also be higher than had been thought. PMID- 9747037 TI - Isolation, tissue expression, and chromosomal assignment of human RGS5, a novel G protein signaling regulator gene. AB - The regulator of G-protein signaling (RGS) proteins have recently been identified as signal transduction molecules which have structural homology to SST2 of Saccharomyces cerevisiae and EGL-10 of Caenorhabditis elegans. Multiple genes homologous to SST2 are present in higher eukaryotes, and the group of these genes is termed the RGS family. RGS proteins are involved in the regulation of heterotrimeric G-proteins by acting as GTPase-activators. A putative new member of the RGS family was isolated from a neuroblastoma cDNA library. The amino acid sequence deduced from the cDNA possessed all consensus motifs of the RGS domain and showed closest homology to mouse RGS5 (90% identical), indicating that it was human RGS5 (hRGS5). The messenger RNA of hRGS5 was abundantly expressed in heart, lung, skeletal muscle, and small intestine, and at low levels in brain, placenta, liver, colon, and leukocytes. The chromosome localization of the gene in the 1q23 region was determined by a monochromosomal hybrid panel and a radiation hybrid panel. PMID- 9747038 TI - Connatal Pelizaeus-Merzbacher disease: a missense mutation in exon 4 of the proteolipid protein (PLP) gene. AB - We investigated the proteolipid protein (PLP) gene in two brothers in a Japanese family with a connatal form of Pelizaeus-Merzbacher disease (PMD). Direct sequencing of the PLP gene revealed an A-to-T transition in exon 4, which led to an Asp-to-Val substitution at residue 202. Their mother was confirmed to be heterozygous for the mutation. The mutation was not found in 78 X-chromosomes of normal Japanese individuals. A correlation between the clinical severity of the disease in the brothers and the Asp202-to-Val mutation in the PLP gene was suggested. PMID- 9747040 TI - Dinucleotide repeat polymorphism in the first intron of the CSR gene. AB - The CSR (cellular stress response) gene encodes a protein that structurally resembles the macrophage scavenger receptor, and is a potent regulator of intracellular reactive oxygen intermediates. We found a polymorphic dinucleotide repeat in the first intron of the CSR gene. This polymorphism will be a useful genetic marker to study diseases associated with oxidative stress. PMID- 9747039 TI - Assignment of the ZIP kinase gene to human chromosome 19p13.3 by somatic hybrid analysis and fluorescence in-situ hybridization. AB - A cDNA for a putative member of the serine/threonine kinase family was cloned from an adult human testis cDNA library. The predicted translation product was identical to ZIP kinase, which has been suggested to play an important role in the induction of apoptosis. The messenger RNA was ubiquitously expressed in various tissues. The chromosomal location of the gene was determined by fluorescence in-situ hybridization and polymerase chain reaction-based analyses with both a human/rodent monochromosomal hybrid cell panel and a radiation hybrid mapping panel. This gene was mapped on the q13.3 region of chromosome 19. PMID- 9747041 TI - Novel polymorphisms in the beta ig-h3 gene. AB - We found three novel polymorphisms in the beta ig-h3 gene in patients with gelatinous drop-like corneal dystrophy: (1) a substitution from CTC to CTT at codon 472 that did not alter an amino acid; (2) a substitution from GCG (Ala) to GTG (Val) at codon 480; and (3) a substitution from C to T in intron 10, three nucleotides upstream from the acceptor site of exon 11. The allelic frequencies of the C:T polymorphism at codon 472 and in intron 10 in the Japanese population were estimated to be 0.778:0.222 and 0.954:0.046, respectively. Although the codon 480 substitution was not observed in 54 unrelated healthy Japanese people, the substation did not co-segregate with the disease phenotype, suggesting that this was a rare, non-deleterious alteration. PMID- 9747042 TI - The optometrist as the patient. PMID- 9747043 TI - The excitement of your profession (Southern California College of Optometry Commencement Address) PMID- 9747044 TI - Proton-beam irradiation of subfoveal choroidal neovascular membranes in presumed ocular histoplasmosis syndrome: a case report. AB - BACKGROUND: Presumed ocular histoplasmosis syndrome (POHS) refers to a choroidopathy that is characterized by the presence of multiple peripheral atrophic chorioretinal scars, peri-papillary atrophy, and choroidal neovascular membranes (CNVM), usually in or adjacent to the fovea. In the United States, POHS is an important cause of loss of central visual acuity in patients between the ages of 20 and 50 years. A number of treatment options for subfoveal and juxtafoveal CNVMs in POHS have been under investigation, including laser photocoagulation, surgical excision of the CNVM, and radiation therapy. CASE REPORT: A 28-year-old women was referred to our office reporting decreased depth perception and finger-counting vision in the right eye for the duration of 1 month. A diagnosis of POHS with subfoveal CNVM was made and the patient was referred for an experimental protocol of proton-beam irradiation. Four months after her initial visit, the patient returned, reporting blurry vision with a blind spot in her left eye. A subfoveal CNVM in the left eye was subsequently treated with irradiation as well. Seven months after the initial treatment, visual acuities were 20/20 in each treated eye. CONCLUSION: Although is currently an experimental procedure, proton-beam irradiation appears to be a promising treatment for subfoveal CNVM in patients with POHS. PMID- 9747045 TI - Ocular prostheses. PMID- 9747046 TI - The enigma of giant cell arteritis: multidisciplinary management of two cases. AB - BACKGROUND: Giant cell arteritis is an enigmatic disease that is characterized by chronic granulomatous inflammation of the walls of large and medium-sized arteries. The process has a predilection for the extradural cranial arteries, which include the ophthalmic and the posterior ciliary arteries. A multisymptom disease of older individuals, giant cell arteritis often manifests challenging issues and diagnostic dilemmas. CASE REPORTS: We illustrate two cases with initial symptoms of intermittent headache, malaise, and decreased visual acuity that were incorrectly diagnosed or ultimately misdiagnosed. The first case represents a patient who was diagnosed as having migraine headache and an erythrocyte sedimentation rate (ESR) that was interpreted by the attending physician as too low to warrant temporal artery biopsy. The second case is that of a patient who had a history of headaches, jaw claudication, and numerous medical evaluations. CONCLUSION: Giant cell arteries is an enigmatic disease with multiple manifestations. The differential diagnoses can range from temporal mandibular joint dysfunction to tension headache. Imminent vision loss as a sequelae of this condition warrants careful review of ocular and constitutional history and prompt treatment. PMID- 9747047 TI - Adult onset foveomacular vitelliform dystrophy: a case report. AB - BACKGROUND: Pattern dystrophies of the retinal pigment epithelium, an arrangement of a pattern of dots, lines, or branches, are infrequent fundus abnormalities. Adult onset foveomacular vitelliform dystrophy (AOFVD) is considered a subtype of pattern dystrophy. Onset occurs during middle age, with an accumulation of yellow gray macular deposits in the deeper retinal layers. Typically electro-oculograms are mildly subnormal or normal. Genetic studies suggest an autosomal dominant inheritance with variable penetrance. CASE REPORT: A case of a 56-year-old Hispanic women with a 1-month onset of "wavy moving vision" in both eyes is presented. Previous ocular and family history were unremarkable. Ophthalmoscopic examination revealed yellow circumscribed subretinal lesions of one-third to one half disk diameter in the foveal centers of both eyes. Fluorescein angiography revealed a circumscribed area of mottled hyperfluorescence surrounding a central hypofluorescent spot. The patient was diagnosed with AOFVD. Examination of the patient's children revealed retinal pigment epithelial disturbances in the two oldest daughters, consistent with pattern dystrophies. CONCLUSION: Our investigation supports an autosomal dominant inheritance pattern, as seen on pedigree. The presence of different pattern dystrophies within the same family suggests a common etiologic continuum. PMID- 9747048 TI - The contribution of glycation to cataract formation in diabetes. AB - BACKGROUND: Despite extensive research, the mechanisms responsible for Type II diabetic cataract formation are still unknown. Recent data have favored non enzymatic glycation. However, the pathways by which hyperglycemia leads to cataract are still unknown. Two possible routes were explored; modification of the lens proteins leading to Advanced Glycation Endproduct (AGE) formation and modification of the ATPase pumps leading to osmotic stress. METHODS: The extent of AGE formation was monitored in fetal bovine eyes using non-tryptophan fluorescence. The amount of carbohydrate bound the proteins was measured after reduction with radiolabelled sodium borohydride. Secondary structure estimations were performed by analysis of data obtained using circular dichroism. The effect of glycation on the Na, K-ATPase ion pumps was investigated by comparing the uptake of radioactive 86Rb in the presence of high concentrations of glucose and fructose. RESULTS: These studies were aimed at determining which of these mechanisms is the more likely route for cataract formation. The first mechanism was examined using two approaches. Firstly, by investigating the effects of increased glucose on the secondary and tertiary structure of lens proteins. Detailed analysis of the structures of the lens proteins in the presence of 200 mM glucose revealed that only alpha-crystallin was slightly affected. More important, however, this change did not lead to any significant aggregation. The second approach involved comparing the mechanism of action and possible benefits of anti-glycating agents. CONCLUSION: It is concluded that Type II diabetes cataracts are unlikely to arise as a result of AGE formation, but rather they form because of disruption of the cells, as a result of osmotic stress, brought about by glycation of the ion pumps. PMID- 9747049 TI - Managed care and eye services to diabetic patients: a study of participant behavior. AB - BACKGROUND: Effective managed care requires the active participation of health professionals and patients alike. The essential elements in management of diabetes involve well-defined professional clinical protocols, adequate patient education, and adherence to these management guidelines by all parties. This study was designed to evaluate how well the involved parties cooperate in this endeavor. METHODS: Health plan records were searched for identified diabetic patients. This information was cross-referenced to vision care claims information. Clinical records of diabetic patients who had an eye examination during the study period were peer-reviewed for evidence of a diabetes notation. RESULTS: Of 2825 diabetic patients identified, we obtained the clinical eye records of 554 for review. Seventy percent of those records noted the presence of diabetes. Ninety-seven percent of the eye examinations included a funds evaluation. Only 56% of records noted any patient education or-- at a minimum- notations about recall advice. CONCLUSION: The nature of diabetes requires that health professionals be more aggressive in the management of individuals at risk or diagnosed with diabetes. PMID- 9747051 TI - Physicians need to stand up to managed care. PMID- 9747050 TI - Clinical diorama. PMID- 9747052 TI - An 'apples-to-apples' comparison of training hours for DOs, PAs, NPs. PMID- 9747053 TI - Need to tailor headache supplement to DOs. PMID- 9747055 TI - Hepatitis C: caution up ahead. PMID- 9747056 TI - Benefits of direct glomerular filtration rate (GFR) determination versus creatinine-based tests for evaluating renal function. AB - To diagnose renal disease earlier and thereby reduce the number of patients with endstage renal disease, accurate, specific diagnostic tests are necessary. The author explains the benefits of using tests that rely on the glomerular filtration rate measurement rather than on traditional creatinine-based diagnostic tests. PMID- 9747057 TI - Pericardial cyst: an incidental finding. AB - Pericardial cysts are rare mediastinal cysts occurring with an incidence of 1 in 100, 000. Characteristically, they occur along the right border of the heart. Their size varies from 1.0 cm to 15 cm, and they are often asymptomatic. Patients with symptoms usually have atypical chest pain. In the case reported here, a 37 year-old man complained of nonproductive cough. Chest x-ray film revealed a pericardial cyst that appeared as a large echolucent unilocular mass along the left border of the heart. Diagnosis was confirmed with the use of both computed tomography and transthoracic echocardiography. PMID- 9747058 TI - Prevention and control of hypertension and diabetes in an underserved population through community outreach and disease management: a plan of action. AB - Hypertension and diabetes are overrepresented in the African-American population and can be particularly devastating in this population. These diseases share genetic predisposition, medical risk factors, and environmental influences as etiologic factors, and they may be interrelated, at least in part, by obesity and accompanying hyperinsulinemia. Noncompliance with treatment plans is a significant barrier to health improvement in both diseases, but increased attention to patient involvement in care is a potential solution to this long standing problem. The Baltimore Alliance for the Prevention and Control of Hypertension and Diabetes was established in January 1998 to promote care to the underserved community of West Baltimore, Maryland, and to improve outcomes of hypertension and diabetes. Based at the University of Maryland School of Medicine, the Baltimore Alliance comprises a community health worker program, a church-based education and screening effort, managed care and pharmaceutical company (Hoechst Marion Roussel) partners, a health policy and services research group, and inpatient/outpatient clinical care sites in the health system. Mobilization, cultural relevance, and partnership are employed to ensure that the Alliance's goals of increased patient enrollment and retention in treatment programs will be achieved. Thereby, improved outcomes--clinical, humanistic, and economic--will result. Novel as well as classic approaches to patient education, compliance, and goal achievement are being pursued. Complete expert systems for hypertension and diabetes disease management are being created and will be implemented in the near future. Baseline practices and current outcomes are being identified to act as historical controls. The organization and administration of the Alliance will serve as a prototype that others may follow. PMID- 9747059 TI - Noncirrhotic portal hypertension in the adult: case report and review of the literature. AB - Noncirrhotic portal hypertension results from thrombosis of the extrahepatic portal vein that subsequently is recanalized. Liver function is preserved. In the adult, esophageal variceal hemorrhage is the most common presentation and may occur years after the portal vein thrombosis. We report the case of a 34-year-old man who presented with recurrent esophageal variceal hemorrhage. After ultrasonographic and angiographic evaluation, a diagnosis of idiopathic noncirrhotic portal hypertension was made. Due to recurrent esophageal variceal bleeding, the patient required surgical intervention to control bleeding. The incidence of noncirrhotic portal hypertension is unknown. Multiple etiologies may cause the disorder, although nearly half are idiopathic. The pathogenesis, clinical manifestations, diagnostic evaluation, natural history, prognosis, and management of noncirrhotic portal hypertension are discussed. Endoscopic management of esophageal variceal bleeding is the preferred therapy. However, when endoscopic treatment fails to control variceal hemorrhage, a distal splenorenal shunt is likely to be the most successful operation. PMID- 9747060 TI - Adult-onset Still's disease associated with G6PD deficiency: a case report and literature review. AB - A 37-year-old man presented with symptoms consistent with adult-onset Still's disease. Fever and leukocytosis were prominent, and the patient was started on high-dose aspirin for possible acute rheumatic fever. He developed severe anemia as a result of glucose-6-phosphate dehydrogenase deficiency. His treatment was changed to naproxen, and he recovered with restoration of his hematologic parameters. Although Still's disease is frequently accompanied by mild-to moderate anemia, the development of severe anemia should raise the possibilities of hemolysis secondary to glucose-6-phosphate dehydrogenase deficiency. PMID- 9747061 TI - Role of an outpatient clinic in screening chronic complications of diabetes: a model for diabetes managed care. AB - The purpose of this study was to evaluate the need for an outpatient clinic for screening chronic complications of diabetes mellitus and to explore the major risk factors for such complications. A total of 558 patients (293 men and 265 women, aged 61.4 +/- 10.0 yr) with non-insulin-dependent diabetes mellitus were recruited. All examinations were performed in all patients except for those with previously known complications. A nonmydriatic fundus camera was used to detect retinopathy. Microalbuminuria was detected with a semiquantitative method. A monofilament, semiquantitative tuning fork and neurometer were used to detect peripheral neuropathy. The relationships of demographic and metabolic factors with diabetic complications were analyzed. Among the 558 patients, 443 (79.3%) were found to have at least one chronic complication. Less than half (41.5%) of patients had been identified as having a complication(s) before screening. The rates of undiagnosed complications ranged from 46.7% to 83.4% for each complication. The duration of diabetes, hemoglobin A1c (HbA1c), and systolic blood pressure (BP) were strongly associated with microvascular complications (p = 0.009, 0.018 and 0.037, respectively). The microvascular complication rates reached a plateau when HbA1c reached 8.0% at least among patients with a systolic BP of less than 130 mmHg. Our findings indicate that undiagnosed complications (average, 58.5%) can be found with routine screening, increasing the chances for prompt attention and early intervention. The duration of diabetes, HbA1c, and systolic BP were strongly associated with microvascular complications. Diabetes care can be improved by the implementation of a screening clinic in daily practice. Identification of the specific risk factors in a defined population in specific clinical settings will allow early modification of interventions for optimal diabetes care. PMID- 9747062 TI - Clinical manifestations and prognostic features of acute methamphetamine intoxication. AB - The prevalence of amphetamine abuse and the frequency of emergency department visits for amphetamine intoxication have increased dramatically worldwide. In this study, we retrospectively investigated the relationship between the prognostic features and clinical manifestations among patients admitted to the emergency department of a university hospital for acute methamphetamine intoxication during a 6-year period. Data collected included gender, age, route of abuse, time between drug exposure and arrival at the emergency department, estimated dose, signs and symptoms, laboratory values, and complications. Emergency therapy and cooling procedures were also recorded. After excluding 26 patients with multiple-drug intoxication, 18 patients (male-to-female ratio, 11:7) were include in the analysis. The mean age was 25.6 years. Thirteen patients survived and five died. Patients who died often presented with coma (80% vs 0%, p = 0.002), shock (60% vs 8%, p = 0.044), convulsions (100% vs 23%, p = 0.007), oliguria (80% vs 0%, p = 0.002), and high body temperature (41.4 +/- 0.5 degrees C vs 39.4 +/- 2.1 degrees C, p = 0.005). Furthermore, patients who died had significantly higher concentrations of blood urea nitrogen (8.7 +/- 2.1 vs 5.6 +/- 2.0 mmol/L, p = 0.01) and serum creatinine (212 +/- 71 vs 115 +/- 27 mumol/L, p = 0.033), and lower values of arterial pH (7.12 +/- 0.12 vs 7.34 +/- 0.10, p = 0.03), than those who survived. In the fatality group, the most common complication was rhabdomyolysis with acute renal failure (5 of 5); multiple organ failure resembling that from heatstroke was the leading cause of death from acute methamphetamine intoxication. In conclusion, the adverse prognostic features in patients with acute methamphetamine intoxication include coma, shock, convulsion, oliguria, and high core temperature. Acidosis, volume depletion, and ischemic renal damage were potential risk factors for development of acute renal failure in these patients. PMID- 9747063 TI - Insulin resistance in patients with Klinefelter's syndrome and idiopathic gonadotropin deficiency. AB - Patients with Klinefelter's syndrome (KS) have hypergonadotropic hypogonadism and have a higher incidence of diabetes mellitus. Patients with idiopathic gonadotropin deficiency (IGD) also have hypogonadism but have no proven impaired glucose metabolism. Because high serum testosterone concentrations are thought to correlate with insulin resistance, we assessed the relationship of testosterone concentration with insulin resistance in patients with KS or IGD and normal subjects. Seven patients with KS, six with IGD, and seven normal individuals (controls) were enrolled. Insulin resistance was evaluated by two methods: the total area under the curve (AUC) and the incremental AUC of serum insulin concentrations in response to a 75-g oral glucose load, and the insulin suppression test. KS patients had significantly higher follicle-stimulating hormone and luteinizing hormone concentrations than the normal controls, while IGD patients did not. The plasma testosterone concentrations were significantly lower in both KS and IGD groups than in controls. The incremental AUC and total AUC were higher in both KS and IGD patients than in normal subjects. The steady state plasma glucose concentrations of the KS and IGD groups were significantly higher than that of the normal group, while the steady-state plasma insulin concentrations were similar in all three groups. After log transformation, the plasma testosterone concentration was negatively related to steady-state plasma glucose concentration in all three groups (r = -0.58, p = 0.019). In conclusion, insulin resistance was consistently noted in patients with KS and IGD. Plasma testosterone concentration is inversely related to insulin resistance. PMID- 9747064 TI - RET protooncogene mutations in patients with apparently sporadic medullary thyroid carcinoma. AB - We examined RET protooncogene mutations in sporadic medullary thyroid carcinoma (MTC), using polymerase chain reaction (PCR)-based sequencing. DNA was extracted from tumor tissue and peripheral blood leukocytes of seven unrelated individuals with apparently sporadic MTC. Oligonucleotide primers were selected to amplify exons 10, 11, 13, 15, and 16 of the RET protooncogene, to examine the sequences of codons 609, 611, 618, and 620 of exon 10, codon 634 of exon 11, codon 768 of exon 13, codon 883 of exon 15, and codon 918 of exon 16. Direct DNA sequencing from PCR products was then performed. The results showed that one patient had a somatic mutation at codon 918 (ATG-->ACG), causing a Met-->Thr substitution. One patient had a de novo germline mutation at codon 634 (TGC-->CGC), causing a Cys- >Arg substitution. Another patient had a germline mutation at codon 634 (TGC- >TTC), causing a Cys-->Phe substitution. In the remaining four cases, no RET mutations were found. Unexpectedly, two offspring of the patient (a female) with a germline mutation at codon 634 (TGC-->TTC) harbored homozygous alleles for the mutation; because the father did not carry this mutation, the other affected allele was suspected to have resulted from a de novo germline mutation of paternal origin. One of these offspring was subsequently diagnosed as having MTC. Our findings suggest that all patients with apparently sporadic MTC should be screened for the RET protooncogene by molecular analysis to detect occult or de novo multiple endocrine neoplasia 2 (MEN 2) or familial MTC. This would allow early treatment of affected family members. PMID- 9747065 TI - Randomized, double-blind, placebo-controlled study of transdermal nicotine patch for smoking cessation. AB - Smoking cessation is an arduous process because nicotine withdrawal syndrome, which occurs following sudden interruption of nicotine use in long-term smokers, frequently prevents them from giving up the habit. Nicotine supplement systems may relieve smokers' nicotine withdrawal symptoms and, thus, help in the process of abstinence from smoking. The purpose of the present study was to investigate the effectiveness and safety of a 30-mg transdermal nicotine patch in a smoking cessation program for Chinese smokers. In this randomized, double-blind, placebo controlled study, 30 heavy smokers, who had smoked more than 20 cigarettes per day for more than a year, were treated with 30-mg transdermal nicotine patches, and 32 heavy smokers were given placebo patches during a 6-week smoking cessation program. The clinical characteristics of the two groups were similar. After 6 weeks, the use of the transdermal nicotine patch was associated with markedly reduced nicotine dependence and severity of withdrawal symptoms. Nineteen (63%, 95% confidence interval, CI, 46%-80%) of the smokers treated with the transdermal nicotine patch had successfully quit smoking at the end of the program (6 weeks) and nine (30%, 95% CI 14%-46%) remained abstinent 1 year later. In contrast, only 11 (34%, 95% CI 18%-50%) of the smokers in the placebo group had successfully stopped smoking after 6 weeks, and three remained abstinent 1 year later (9%, 95% CI 0%-19%). However, there was no statistically significant difference between the two groups of smokers after 1 year of follow-up (p = 0.08). Side-effects were minimal and did not affect the efficacy of the skin patch. The results indicate that the transdermal nicotine patch is an effective aid in smoking cessation programs. PMID- 9747066 TI - Optimal timing of retina examinations for premature infants. AB - Developments in modern neonatal intensive care have resulted in increased survival of very premature infants. Along with this increase in survival, there has been a concomitant increase in the frequency of retinopathy of prematurity (ROP). We investigated the relationship between the severity and the time course of ROP as well as the optimal time for retinal examinations of premature infants of various birth weights and gestational ages. A total of 80 premature infants were enrolled for analysis. The mean postnatal age of infants at the time of diagnosis was 7.3 +/- 3.4 weeks for stage 1 retinopathy, 9.2 +/- 5.8 for stage 2 retinopathy, 9.5 +/- 3.8 for stage 3 retinopathy, 10.7 +/- 3.7 for threshold disease, and 11.7 +/- 3.2 for stage 4 retinopathy. The mean postconceptional age of infants at the time of diagnosis was 35 +/- 3 for stage 1 retinopathy, 36.4 +/ 3 for stage 2 retinopathy, 37.6 +/- 3.5 for stage 3 retinopathy, 38.4 +/- 3.5 for threshold disease, and 40 +/- 3.7 for stage 4 retinopathy. The age at the time of the initial detection of ROP was from the ninth to 10th week among infants weighing less than 1,000 g at birth and from the fifth to seventh week among those weighing 1,000 g or more at birth. However, the postconceptional age at the time of initial detection of ROP for the whole group was 36 weeks and was not influenced by birth weight or gestational age. Therefore, we suggest that postconceptional age, rather than postnatal age, should be used to decide the timing of retinal examinations for premature infants. PMID- 9747067 TI - Anesthesia for pediatric herniorrhaphy or hydrocelectomy: comparison of propofol/ketamine and thiopentone/halothane. AB - Total intravenous anesthesia has recently become available for ambulatory surgery. It has the advantages of decreased air contamination from volatile anesthetics and decreased exposure of operating room personnel to volatile anesthetics. The purpose of this study was to compare the anesthetic properties of propofol/ketamine (total intravenous) anesthesia and thiopentone/halothane (intravenous and gaseous) anesthesia for herniorrhaphy or hydrocelectomy in children. Sixty children aged 2 to 7 years scheduled for herniorrhaphy or hydrocelectomy were allocated to two groups. The propofol/ketamine group (group 1) received a loading dose of intravenous propofol 3 mg/kg followed by propofol infusion 200 micrograms/kg/minute; additional bolus doses of propofol 1 mg/kg were given as needed or the infusion dose was increased or decreased by 33 micrograms/kg/minute as needed. Ketamine 1 mg/kg was administered intravenously 2 to 3 minutes before herniorrhaphy or hydrocelectomy to reinforce the analgesic and anesthetic effects of propofol. The thiopentone/halothane group (group 2) received intravenous thiopentone 6 mg/kg followed by halothane with 40% oxygen using a mask. Group 2 patients maintained spontaneous breathing with intermittent assistance and group 1 patients maintained spontaneous natural airway breathing during anesthesia. The scores on the postoperative assessment scale were higher in group 2 patients, indicating poorer anesthesia recovery characteristics, but the differences were not significant. Pain on injection was more frequent in group 1 (12/32) than in group 2 (2/28). The incidence of vomiting in group 2 (6/28) was significantly higher than in group 1 (0/32). We conclude that propofol/ketamine allows patients to maintain spontaneous natural airway breathing during anesthesia, and its analgesic and anesthetic effects are comparable to those of thiopentone/halothane. Propofol/ketamine is appropriate for pediatric herniorrhaphy and hydrocelectomy. It can be recommended for pediatric ambulatory surgery. PMID- 9747068 TI - Intramedullary versus extramedullary tibial alignment guides in total knee arthroplasty. AB - The intramedullary alignment guide is superior to the extramedullary guide for preparation of the femur in total knee arthroplasty. However, there is controversy over which guide is more appropriate for the tibial sector. We retrospectively compared the accuracy of the intramedullary and extramedullary guides for tibial cutting in patients undergoing total knee arthroplasty. Total knee arthroplasty was performed in 100 knees (68 patients) during a 2-year period. The intramedullary rod was used for preparation of the femur in all cases. For the tibia, each guide system was used in 50 knees. The intramedullary rod was not used in tibias with extreme deformity where the rod could not pass at least two-thirds of the way through the medullary canal. Standing anteroposterior radiographs of the hip to the ankle were taken before surgery and 2 to 6 months postoperatively. The angle formed by the intersection of the tibial mechanical axis and the undersurface of the tibial component (tibial component angle) was measured to check the accuracy of the tibial alignment system. We found no significant differences in the mechanical axis, tibiofemoral alignment, or the tibial component angle between the two groups. The proximal tibial cuts were within 2 degrees of the ideal (90 degrees) in 84% of knees treated with the intramedullary guide, and in 82% of those with the extramedullary guide (p > 0.1). These findings suggest that both guide systems can yield satisfactory alignment. If the tibia is not badly deformed, the intramedullary rod can produce tibial cuts as accurately as the extramedullary system. PMID- 9747069 TI - Cat-scratch disease caused by Bartonella henselae: the first case report in Taiwan. AB - We report a typical case of cat-scratch disease caused by Bartonella henselae, in Taiwan. A 20-year-old man developed right axillary lymphadenopathy 2 weeks after being scratched on his right hand by a kitten. The axillary lymphadenopathy resolved gradually and spontaneously after 10 weeks without specific treatment. Serologic tests were not done during the acute stage of the event. However, an immunofluorescent antibody test performed during the convalescent stage was positive for B. henselae antibodies, and the concentration dropped by fourfold 2 months later. Histopathologic examination of a biopsy specimen from the right axillary lymph node revealed findings characteristic of cat-scratch disease including multiple foci of microabscesses surrounded by histiocytes and infiltration by plasma cells and lymphocytes. This is the first reported case of cat-scratch disease in Taiwan, with a history of contact with a cat, a positive serologic test for B. henselae infection and characteristic histopathologic findings of cat-scratch disease which met the criteria for diagnosis. PMID- 9747070 TI - Lung cancer in pregnancy: report of two cases. AB - Lung cancer during pregnancy is rare, although the number of case reports has been increasing in recent years. Herein, we describe two cases of lung carcinoma complicating pregnancy with different presentations and outcomes, and review the relevant literature. The first case involved a 31-year-old patient with squamous cell carcinoma with multiple bone metastases. The initial symptoms were productive cough and dyspnea on exertion during the second trimester of pregnancy, to which the patient paid little attention. Chemoradiation was started 1 month postpartum, soon after the diagnosis was made, but with little response. She died at home several days after palliative radiotherapy. The second case involved a 34-year-old patient with poorly differentiated lung carcinoma with brain metastasis. Left hemiparesis had developed initially during the third trimester. She underwent excision of the metastatic brain tumor and received radiotherapy to the left lung tumor and brain. The patient is still alive after a follow-up period of more than 1 year. Delayed diagnosis may be the main problem in the management of lung cancer during pregnancy, because of misinterpretation of common respiratory symptoms and physicians' reluctance to use radiologic imaging studies owing to concerns over the safety of the fetus. Thus, we suggest chest radiographs with abdominal lead shielding for pregnant patients with protracted cough and hemoptysis. Treatment of unresectable lung cancer during pregnancy generally consisted of radiation therapy with or without chemotherapy in previous reports, but the optimal therapy is still unknown, owing to inadequate case numbers and insufficient follow-up data. PMID- 9747071 TI - Intestinal obstruction and peritonitis resulting from gastrointestinal histoplasmosis in an AIDS patient. AB - Gastrointestinal histoplasmosis complicated by intestinal obstruction and peritonitis has not been reported. We report a case of gastrointestinal histoplasmosis in a 27-year-old patient with acquired immunodeficiency syndrome (AIDS). The patient was a Chinese man from Thailand with a history of intravenous drug use and unprotected sex with female prostitutes. He was admitted for prolonged fever, abdominal pain, and diarrhea. Colonoscopy revealed volcano-like ulcers and tumors, while computed tomography of the abdomen showed a colon tumor and hypoattenuated lymphadenopathy of the retroperitoneum. Histopathologic examination as well as cultures of colon biopsy specimens and an aspirate from the retroperitoneal lymphadenopathy revealed Histoplasma capsulatum. Intestinal obstruction and peritonitis requiring surgical intervention developed, despite amphotericin B therapy. Histoplasmosis should be included in the differential diagnosis in AIDS patients who present with colon tumors, retroperitoneal lymphadenopathy, and peritonitis. PMID- 9747072 TI - Drug resistance patterns of tuberculosis in Taiwan. AB - To evaluate the patterns of drug resistance of Mycobacterium tuberculosis in Taiwan, a total of 1,091 isolates collected from patients from January 1996 through December 1996 were tested for drug susceptibility using the absolute concentration method at the Taiwan Provincial Chronic Disease Control Bureau. The overall drug rate of resistance to at least one drug was 35.5%. Among the 249 isolates from patients who had never been treated for tuberculosis, 16.1% were resistant to one or more drugs; 1.6% were resistant to at least isoniazid and rifampin. Of 200 patients with prior antituberculosis treatment, 67.0% had isolates resistant to one or more drugs and 46.0% had isolates resistant to at least isoniazid and rifampin. We conclude that drug-resistant M. tuberculosis is an important issue in tuberculosis treatment in Taiwan, especially when dealing with patients with a prior history of antituberculosis treatment. More aggressive interventions, such as directly observed therapy, short-course, are needed to improve the cure rate of pulmonary tuberculosis and to decrease resistance rates. PMID- 9747073 TI - Facing challenges: managed care and HCFA. PMID- 9747074 TI - The medical record is not a billing record. PMID- 9747075 TI - Aetna US Healthcare responds. PMID- 9747076 TI - MAG fights to revamp E & M codes. PMID- 9747077 TI - Contracting in the managed care market: a defining challenge for organized medicine. PMID- 9747078 TI - Prison health: a matter of priority. PMID- 9747079 TI - The death of Thomas Wolfe: a 60-year retrospective. AB - It seems likely that an old tubercular lesion of the lung had been activated during Wolfe's acute pneumonia and that the disease had spread to the brain. The chest x-ray with the right upper lobe infiltrate was certainly typical of pulmonary tuberculosis, and the spinal fluid findings were characteristic of tuberculous meningitis with 230 cells, 75% of which were mononuclear, which Dandy felt "practically made the diagnosis." Tuberculosis was so prevalent worldwide in the early 20th Century, around the time of Wolfe's birth, but began to decline dramatically in the early 1950s with the introduction of modern chemotherapy and isoniazid (INH). In the U.S. the number of reported cases of TB decreased from 84,304 to in 1953 to 22,201 in 1985. Unfortunately, however, the number of cases has been increasing since 1985, due to a variety of factors including decline in public funding for TB control, HIV infection, immigration, homelessness, substance abuse, and incomplete therapy. Today, Wolfe would have been easily treated and probably cured. Would earlier diagnosis of his condition have made any difference in the outcome? The answer is uncertain, since sanatorium therapy (rest and environment) and surgery were the only available treatments at the time of his illness. Had he lived in a later generation, he might not have met his death at the age of 37. PMID- 9747080 TI - Lemonade out of lemons--tinnitus retraining therapy. PMID- 9747081 TI - Ten indications for the electron beam CT scan of the coronary arteries. PMID- 9747082 TI - Health care corporate compliance programs. PMID- 9747083 TI - Our own science. PMID- 9747084 TI - Should ability to pay influence provision of epidural analgesia/anesthesia to women in labor? PMID- 9747086 TI - Being prepared for a pregnant code blue. AB - Cardiopulmonary arrest is rare in pregnancy. To provide the most proficient care to a pregnant woman requiring cardiopulmonary resuscitation, nurses must first understand the physiologic changes that normally occur in pregnant women. This article reviews the physiologic adjustments made by the human body during pregnancy and the manner in which these can affect efforts during resuscitation. Preparation and organization are essential in implementing care during resuscitation of a pregnant woman, which is a crisis situation for everyone involved. PMID- 9747085 TI - Prevalence of feeding tube placement errors & associated risk factors in children. AB - PURPOSE: The purpose of this study was to determine the prevalence of feeding tube placement errors in children. DESIGN AND METHODS: The hospital records of 201 children having both an enteral tube and at least one radiograph showing tube placement were retrospectively reviewed. Chart review was also used to determine the risk factors associated with these errors. Tube placement error was defined as tube tip or orifices in the esophagus or intestine (if the tube was supposed to be in the stomach) or tip or orifices in the esophagus or stomach (if the tube was supposed to be in the intestine.) RESULTS: On the first day, a radiograph documenting tube placement showed that 32 of the 201 children (15.9%) had tube placement errors. Overall, 53 tube placement errors were evident during the 385 observation days on which radiographs were obtained (13.8%). Of the 201 children, 42 (20.9%) had experienced tube placement errors at some time during the period reviewed. Over all radiograph days, activity level was independently related to radiographic tube placement (p = < 0.02), with more errors among active children. Also, classification regression tree analysis showed that age, level of consciousness (alert or comatose versus semicomatose), abdominal distention, vomiting, and orogastric tubes were associated with more tube placement errors. NURSING IMPLICATIONS: Nurses need to be especially careful in assessing tube placement if the pediatric client has one or more of the identified risk factors. Health care providers need to carefully weigh the benefits and risks of feeding by nasal or oral enteral tubes versus the benefits and risks of feeding by endoscopically or surgically placed enteral tubes. PMID- 9747087 TI - Perinatal hepatitis B: update & recommendations. AB - The hepatitis B virus (HBV) is the most prevalent chronic infectious disease in the world, and should be better understood by nurses caring for families. Perinatal acquisition is the major cause of infection in infants and children. Without vaccine during infancy, 90% of infants born to women positive for the virus will go on to become lifelong carriers. There are significant sequelae associated with HBV infection, ranging from fulminant HBV to chronic liver disease to an increased risk for carcinoma. A comprehensive prevention and treatment strategy has been developed by the Centers for Disease Control and Prevention, which includes screening of all pregnant women for the presence of HBV, the administration of hepatitis B immunoglobulin (HBIG) at birth, and the administration of hepatitis B vaccine at birth, at 1 month of age, and at 6 months of age. Nurses working in the perinatal and pediatric specialties must understand the implications of HBV to help prevent transmission and to assist in the coordination of care and advocacy for affected populations. The community health implications for the care of women and children with HBV are clear, giving nurses the opportunity to develop a closer linkage between hospital- and community-based nursing practice. PMID- 9747088 TI - A review of research instruments for use during the postpartum period. AB - Locating existing instruments available to measure key variables is critical to the success of a research study. However, this process can be a time-consuming activity. In this article, selected instruments available for use in research during the postpartum period are reviewed. These instruments are divided into two categories. The first category deals with instruments that focus on mothers only. These questionnaires measure mothers' perceptions of their deliveries, their adaptation to motherhood, and their breastfeeding experiences. The second category centers on instruments that both parents can complete. These instruments measure parents' perceptions of their parenting role, sense of competence, problem-solving ability, and perceptions of their newborns. The psychometric properties of each instrument are described, and selected nursing research studies that used the instrument are reviewed. PMID- 9747089 TI - Proactive nursing: the evolution of a task force to help women with postpartum depression. AB - In response to several women who presented with postpartum depression in 1 year, a group of nurses developed a task force made up of hospital nurses, obstetricians, psychiatrists, pediatricians, family practitioners, lactation specialists, home care nurses, and mental health counselors. The purposes of this task force were to educate health care professionals about postpartum depression, to help identify women who might be affected, and to develop interventions for adjusting to parenthood. This article details the evolution of that task force, and how it has assisted not only the women but also the health care providers involved. PMID- 9747090 TI - Expanding the newborn screen: terrific or troubling? AB - Amid controversy about cost-benefit ratios and ethical issues of discrimination and presumed consent, individual U.S. states are on the verge of expanding the newborn screen. The success of population-based newborn screening for genetic and metabolic disorders has been called the miracle of our times. Rapid scientific growth in genetic mapping and laboratory testing has resulted in increased genetic testing in the adult population as well. Caution must be exercised, however, before mass population testing of newborns is considered. Proposed testing for treatable but incurable conditions such as cystic fibrosis and human immunodeficiency virus (HIV) remains controversial. Health professionals must meet the challenge of educating themselves and others in order to advocate for children and families. Their efforts should extend to the legislative arena where decisions to amend the newborn screen are made. Nurses, genetic counselors, and other health professionals are in key positions to conduct research in this area to expand knowledge about the implications of genetic testing for the families they serve. PMID- 9747091 TI - Ceftriaxone sodium. PMID- 9747092 TI - Bed rest in complicated pregnancy. PMID- 9747093 TI - Maternal serum alpha-fetoprotein screening. PMID- 9747100 TI - [Changes in and acceptance of surgical and noninvasive therapy procedures in cholecystolithiasis]. AB - BACKGROUND: The treatment of cholecystolithiasis has changed fundamentally in recent years due to the development of non-surgical techniques (extracorporeal shockwave lithotripsy [ESWL], oral litholysis) and the implementation of laparoscopic cholecystectomy. PATIENTS AND RESULTS: Retrospective analysis of 2270 patients (1649 women, 621 men; age: 47.2 +/- 14 years) presenting with gallstone disorders in a university medical outpatients department between 1988 and 1992 in order to be instructed as to the most suitable therapy method bear witness to the rapid change in therapeutic procedure. Laparoscopic removal of the gallbladder has virtually supplanted conventional cholecystectomy, and within 5 years the proportionate role of ESWL has declined from 21 to 12%. Over the years, the proportion of patients requiring no therapeutic intervention remained constant (at about 20%). The therapeutic recommendations of the "experts" were implemented in almost 80% of cases. The majority of patients were satisfied with the chosen therapeutic approach (surgery: 93.0%, ESWL: 77.6%), although 44% of ESWL-patients and 36% of surgically managed patients reported complaints which persisted even after completion of therapy. Despite unsuccessful ESWL (residual fragments or recurrent stones) 58/95 (61%) of interviewed patients would again give preference to this non-invasive modality in the event of a renewed therapeutic decision. CONCLUSION: Only a few years after its introduction, laparoscopic cholecystectomy has asserted itself as the predominant treatment option. But as far as acceptance and preference by the patient are concerned extracorporeal shockwave lithotripsy--as a non-invasive treatment modality--also enjoys high popularity and can be recommended as an alternative to surgery in suitable patients chosen according to the currently established stringent selection criteria. PMID- 9747101 TI - [Chemotherapy of alveolar echinococcosis with benzimidazoles. A prospective long term study]. AB - BACKGROUND: Mebendazole and albendazole are the drugs of choice for the treatment of alveolar echinococcosis. In this prospective study we present and evaluate the outcome of the long-term treatment with both drugs. PATIENTS AND METHODS: Forty four patients were treated with either mebendazole or albendazole and they were followed up for an average of 42 months. Success of treatment was defined as non progression for more than 1 year. RESULTS: The overall success-rate was approximately 80% (35/44). An initial regimen was recurrence-free in 64% of cases under mebendazole and in 73% of cases under albendazole. Half of the cases with recurrent disease could be stabilized after changing the therapeutic regimen. Seven patients received a continuous regimen with albendazole. They were observed over an average of 19 months without signs of progression nor significant side effects. CONCLUSION: This open-labelled prospective study demonstrates the high therapeutic efficacy of both mebendazole and albendazole with similar response rates in the treatment of alveolar echinococcosis. In Germany, serum levels for mebendazole can easily be obtained at numerous institutes, while serum levels for albendazole are rarely available. On the other hand, albendazole reduces costs by over 40%. A simplified mode of intake and a reduced number of side effects argue in favor of the preferred use of albendazole. Albendazole in alveolar echinococcosis is only licensed for intermittent application. Nonetheless, continuous treatment may be considered in inoperable cases or progressive disease. PMID- 9747102 TI - [Economic evaluation of various hepatitis B vaccination strategies in children and adolescents]. AB - AIM: In this cost-effectiveness study 4 different vaccination strategies against hepatitis B in children and adolescents are evaluated and compared with the situation without immunization. EXAMINATION: Projections are made for the population of the today's adolescents underage 15 and the newborns of the next 30 years. The number of avoided hepatitis B virus (HBV) infections and the cases of disease as well as the costs associated with treatment and vaccination are determined. The course of incidence of the hepatitis B virus is observed for different age groups. RESULTS: Compared to the situation without any vaccination against hepatitis B, a decrease of the remaining infections of at least 18,900 up to 46,600 could be expected during the next 30 years. The treatment costs for the remaining cases of disease could be reduced by 0.4 up to 1.6 billions DM. The remaining expenditures for treatment and vaccination would be limited to 2.3 up to 3.4 billions DM. The net costs of a vaccination are determined as about 14,200 up to 63,000 DM per avoided case of infection. Considering the commonly accepted number of unreported cases of hepatitis B as to be the 5- to 10 fold of the known incidence, all of the 4 compared vaccination strategies will be cost-effective and associated with net savings of about 5,900 up to 36,400 DM per avoided case of hepatitis B virus infection during 30 years. The epidemiological situation will be positive influenced by such a mass vaccination. The minimization of incidence is shown for the different age groups. CONCLUSION: Considering these economical arguments, first the vaccination of all adolescents between age 11 to 15 and second the vaccination of all children/adolescents between age 0 to 15 are the preferable strategies. The immunization of all children/adolescents between age 0 to 15 is the most effective strategy from an epidemiological point of view. PMID- 9747104 TI - [Concept of stress ulcer prevention. Is re-thinking necessary?]. AB - BACKGROUND: The efficiency of stress ulcer prophylaxis in the prevention of gastrointestinal bleeding in critically ill patients has led to its widespread use. The lower incidence of stress ulcer bleeding, the side-effects and the cost of the prophylaxis have made it necessary targeting this preventive therapy to those patients most likely to benefit. Metaanalysis of studies on patients who received no stress ulcer prophylaxis showed few critically ill patients with important gastrointestinal bleeding. INDICATIONS: Patients who benefit most from receiving stress ulcer prophylaxis are critically ill patients with coagulopathy, or those requiring mechanical ventilation for more than two days. In patients with headinjuries, widespread burns or severe hypotension, the effects of stress ulcer prophylaxis have not been fully researched, but we would recommend administering stress ulcer prophylaxis in these cases. TREATMENT: Following a recent metaanalysis, stress ulcer prophylaxis is performed either with H2 blockers (ranitidine, famotidine) or sucralfate. PMID- 9747103 TI - [Leptin--new knowledge on the pathogenesis of obesity]. AB - Cloning of the ob-gene and characterization of its gene product leptin has led to the identification of a satiety factor, which signals the amount of peripheral fat stores to the central nervous system and regulates further feeding behaviour, thus playing a central role in the regulation of body weight. Soon after cloning of the ob-gene, a leptin-binding receptor has been identified in the central nervous system as well as in various peripheral organs. A feedback loop between peripheral fat stores and leptin receptors in the central nervous system appears to play an important role in normal body weight regulation. In contrast to human obesity, which associated with leptin resistance of uncertain etiology, the obesity syndromes associated with several animal models are now known to result from the interruption of the feedback loop at different points. Moreover, leptin may play a role in manifestation of insulin resistance and type II diabetes. Since the identification of leptin, a vast number of studies have been conducted to assess the molecular mechanisms and signal transduction pathways that are involved in the development and manifestation of obesity. From the large body of data generated to date, novel concepts of the regulation of energy balance and target strategies to control human obesity should soon be forthcoming. PMID- 9747105 TI - [Vulvar involvement as a rare extra-intestinal manifestation of Crohn disease]. AB - BACKGROUND: Crohn's disease may involve all parts of the gastrointestinal tract. Extraintestinal manifestations with and without continuity to the intestine are described. The most common complications are the involvement of bone (articulations), liver and eyes. Crohn's disease of the vulva is rare and only a few cases have been reported in the literature. CASE REPORT: The 45-year-old woman has a 15-year history of a marked chronic-inflammatory bowel disease. The diagnosis of Crohn's disease was never confirmed histologically. In 1984, she developed a painful vulvar swelling and erythema. A fungal infection was suspected and she was treated with topical antimycotic medication, but there wasn't any improvement. In July 1995, biopsies of the vulva showed a granulomatous inflammation consistent with Crohn's disease. Oral metronidazol therapy (20 mg/kg/day) was started. After 2 months, the erythema had healed completely but the vulvar swelling remained. CONCLUSIONS: The treatment of this extraintestinal manifestation is very difficult. Systemic or topical application of steroids is without response in most cases. Surgical excision showed good results in a few cases. A long-term treatment with oral metronidazol 20 mg/kg/day could be a chance for healing without adverse effects. PMID- 9747106 TI - [Postinfectious bilateral leg edema and lumbar sciatica]. PMID- 9747107 TI - [Significance of philosophy, bio-mathematics, biometry and model theory for medicine]. PMID- 9747108 TI - [Why?--Philosophical difficulties with the causal question]. PMID- 9747110 TI - Aspects of 1,25-dihydroxyvitamin D3 binding sites in fish: an autoradiographic study. AB - The distribution of specific binding sites for vitamin D3 in adult female and male Xiphophorus helleri is studies after injection of tritiated 1,25 dihydroxyvitamin D3 (vitamin D) by thaw-mount autoradiography. Five hours after injection of labeled vitamin D specific nuclear binding is present in brain, pituitary, skin, gills, cartilage, gut, liver, pancreas, spleen, kidney, muscle, ovary, and testis. Cytoplasmic binding exists strongest in gills, gut, and kidney while it is comparatively weak in hepatocytes. In reproductive organs cytoplasmic retention of radioactivity is also present in oocytes. Weak nuclear labeling exists in interstitial cells in ovary. Conspicuous nuclear labeling exists in active lobules of testis, while inactive lobules show occasionally a few labeled cells. The results demonstrate specific binding and retention of vitamin D in many target organs of teleost fish, suggesting an extensive and multifunctional regulatory role of this steroid hormone of sunlight. PMID- 9747109 TI - The role of reparative processes in teratogenesis. AB - Introducing the review article the multiple relations between reparative processes and prenatal development are briefly discussed. In continuation the theoretical framework of possible and plausible repair in teratogenesis is outlined and documented by some, so far existing direct and indirect examples from experimental and human anatomo-clinical observations. The theoretical and practical importance of the problem and the still open questions to be answered by further investigations are mentioned. PMID- 9747111 TI - Cellular and subcellular antioxidants involved in liver susceptibility to xenobiotics. AB - Our previous studies have shown that some important serum erythrocyte and liver antioxidative defense factors--ceruloplasmine, catalase, superoxid dismutase, glutathione -SH and protein-SH groups--were sensitively affected by xenobiotics impact in rat. The data are completed here with the subcellular level of these factors. Dose or time exposure induced appreciable changes of the antioxidative factors (catalase, superoxid dismutase, ceruloplasmine) suggesting their redistribution between the subcellular fractions. The liver morphological investigations in all experimental variants express the aggressivity of Carbofuran administered alone and the "correctional" effects of the modulators' interference. PMID- 9747112 TI - Ethanol stimulates the formation of free oxygen radicals in the brain of newborn rats. AB - The possibility of free radicals effects in ethanol-induced teratogenesis was investigated by determining the production of reactive oxygen species (ROS) in the neonatal rat brain. Ethanol 33% was administered daily, by i.p. injection from day 8 of pregnancy to day 6-8 p.n. The presence of the lipid peroxidation process (indicating ROS formation) was determined by using a qualitative and quantitative analysis of malondialdehyde (MDA). An important increase of MDA was found suggesting the involvement of ROS in the pathogenetic mechanism of alcohol embryo- and fetopathy. PMID- 9747113 TI - Electronmicroscopic observations regarding the presence of natriuretic granules in the ventricle of patients with cardiopathies. AB - As part of studies regarding the human myocardium biology in hereditary and acquired cardiopathies, we showed the presence of natriuretic granules (NG) not only in atriums but also in ventricles, in ten of our cases with intraoperative myocardium biopsies. Ultrastructurally, the natriuretic granules occur in lesions produced by hemodynamic and consecutively hypoxic disturbances, being present at the ventricular level, too, both near the Golgi apparatus that secrets them, and diffusely, in cardiomyocytes, beneath the altered organelles and phagolysosomes. Their aspect in similar in cardiac malformations (DSIA. Fallot tetralogy) and in mitral valvulopathies, their abundance being in connection with congestive heart failure (CHF). Although predominant in CHF class I and II, they occur also in the severe decompensations class III and IV, being perhaps felt at the cardiac level as they are released in the blood. This phenomenon is expressed by electronmicroscopic presence of natriuretic granules in the subsarcolema and the increased plasma level of natriuretic peptide, according to biochemical findings reported in the literature. They produce a vasodilating, diuretic, natriuretic effect, contributing to blood pressure regulation and testifying in the neuroendocrine role of human myocardium. PMID- 9747114 TI - Immunocytomorphological study on the pathogenesis of ankylosing spondylarthritis. AB - The present investigation is based on the cytomorphological, histopathological (HE, VG, PAS-Alcian, Safranin 0, Gomori), histoenzymological (acid phosphatase, chondroitinsulphatase, peroxidase) and immunological (rheumatoid factor (RF), circulating immune complexes (CIC), anticolagen II antibodies and C reactive protein (CRP) study on ankylosing spondylarthritis (2.5 cases). The synovial fluid (SF) synoviocytogram showed cytosis (6.067/mm3), with polynucleosis (65.19%) and ragocytosis (17.73%) as compared with the hydrarthrosie SF characterized by lymphocytosis (47%). Enzymological findings revealed phosphatasic and myeloperoxidasic activity in the ragocytary polymorphonuclear (PMNs) and mononuclear cells. Histopathologically, the severe forms of AS correlated with villous chronic synovitis, associated to processes of obliterating vascularitis, fibrosclerosis, necrosis and calcification of disintegrated synovial structures. The articular cartilage was severly damaged, while osseous necrobiosis was noted at the osteocartilaginous junction. Histoenzymologically, the chondrocytes and synovial macrophages showed lysosomal and oxidative enzymatic activity. Immunological assessments (72 sera and 25 synovial fluid samples) showed pathological values of circulating immune complexes, anticollagen antibodies and C reactive protein. Correlation of immunocytomorphological findings demonstrates the involvement of immunologic and enzymatic factors in the pathogenesis of AS. PMID- 9747115 TI - Cytologic diagnosis of transitional cell carcinoma of the urinary bladder. Comparison with endoscopical and pathological findings on 538 cases. AB - There were investigated 583 cases with tumors of the urinary bladder and 612 patients with non-malignant diseases of the urinary tract. Samples of voided urine were taken from all cases and direct smears fixed by drying were stained by rapid blue polychrome-tanin Dragan method Cytological results were compared with endoscopical and pathological findings. The overall rate of real positive results was 91.7% and false negative results were noticed in 8.3% of cases. A direct relationship between real positive results and histological "G" was found. Causes for false negative results were: tumor developed in a bladder diverticulum, calcified tumor, irradiated tumor, insufficient quantity of voided urine, chronic urinary infections and underestimation of cytological criteria of cellular malignancy. There were 9 false positive results in patients with nonmalignant diseases, due to lithiasis, chronic renal failure and chronic urinary infections. The cytological grade of differentiation was performed by the method purposed by Friedman and Ash, and concordance with the standard histological finding was 76.4%. Urine cytology is thought to be a useful method in the primary diagnosis and recurrences of transitional cell carcinoma of the urinary bladder, in all patients with hematuria, recurrent infections of the urinary tract and neglected lithiasis. PMID- 9747116 TI - Computer-based image analysis of nucleoli in prostate carcinoma. AB - The authors have evaluated the nucleoli in 37 cases of prostatic carcinoma. The increase in Gleason's grade 1 correlated with an increase in the percentage of nucleolated nuclei. The increase in the percentage of bi- and trinucleolated nuclei was accompanied by a decrease of the uninucleated nuclei. The values of morphometrical parameters studied in the individual nucleoli (area, perimeter, diameters) increased in Gleason grade 2 tumors as compared with grade 1 tumors. In tumors higher grades there was, however, a progressive decrease of these values. PMID- 9747117 TI - Histopathologic and immunohistochemic correlations in virus B chronic hepatitis. AB - Complete diagnosis of chronic hepatitis relies on exploring the liver by bipsic punction, performing the classic histopathologic and immunohistochemic exams. We worked out viral antigens hepatocytes by using avidin-biotin-peroxidase complex technique as following: Ag HBs placed in cytoplasm or at the level of the cell membrane. Ag HBc preferably placed in nucleus and, a small part of it, in cytoplasm. Ag HD present especially in nucleus. A correlation between tissular antigen expression and hepatic histopathologic aspect was established. Two main types of viral expression were remarked: a regressive type reflected by cytoplasmatic Ag HBs in the absence of generalised nuclear Ag HBc--situations linked to persistent chronic hepatitis: an aggressive type characterised by the presence of the focal nuclear Ag HBc, cytoplasmatic Ag HBc or antigen HD- situations linked to active chronic hepatitis with various degrees of severity. PMID- 9747118 TI - Microscopic and ultrastructural aspects in retinoblastoma. AB - A number of 12 retinoblastoma cases were studied. We had in view a number of histopathological aspects (tumor type, neighbouring structures invasion) which are considered to be decisive in appreciating the vital prognosis. We mention that undifferential retinoblastoma of histopathological type was met in 8 cases and the neighbouring structures invasion was present in 7 of them. We also studied the ultrastructura of the tumor cells belonging to the malignant tumor. PMID- 9747120 TI - Immunohistochemical markers in the morphological diagnosis of lung carcinoma. AB - Conventional histopathological criteria based on light microscopy are used in pulmonary oncologic pathology in order to establish the diagnosis of tumor, but most frequently they are insufficient, accurate diagnosis requiring ultrastructural and immunohistochemical investigations. The method of immunostaining allowed some molecular marker to be evaluated. Some of them seem to be important in carcinogenesis as a general process, while others have high specificity for lung tumors. Estimation of EGFR and c-erbB-2 protein immunoreactivity showed a significantly stronger staining with NSCLC and was correlated to the poor differentiation of the tumors, undergoing an aggressive biological behavior and an unfavorable prognosis. The expression of p53 protein was found in 19 cases by immunostaining with DO-7 antibody. Immunotracing of more than 50% of the tumoral cells was a predictive factor for the progression of the disease. The growing rate of tumoral proliferative activity was evaluated by immunotracing technique (MIB-1), allowing the Ki-67 index of labeling to be calculated. PMID- 9747119 TI - Considerations on endometrial epithelial metaplasia. AB - Our study aimed to the various histopathological aspects of the endometrial epithelial metaplasia and the endometrial lesions which appear more frequently associated to these changes. Being considered as isolated, endometrial epithelial metaplasia has no prognostic relevance. It is important that the histopathologist recognize it, because its association to some benign lesions (especially to the endometrial glandular hyperplasia) may generate confusion with carcinoma. PMID- 9747121 TI - Histologic criteria for the diagnosis of gastric low-grade malt lymphoma. AB - Low-grade B cell lymphoma of the stomach have the features of mucosa-associated lymphoid tissue (MALT) and is characterised by lymphoid hyperplasia and infiltrations of lymphocytes into glandular epithelium or lymphoepithelial lesions. It may be difficult to distinguish low-grade lymphomas from benign inflammatory lymphoid infiltrates. Nine primary gastric lymphomas and 20 benign lymphoid hyperplasia were investigated for the type of tumour cells, lymphoepithelial lesions. Dutcher bodies, cytologic atypia, the density of lymphoid infiltrates, invasion of the muscularis mucosae, germinal centers, reactive epithelial atypia and acute inflammation. Our results suggest that B cells proliferation, lymphoepithelial lesions, cytologic atypia and Dutcher bodies are very important for the diagnosis of low-grade MALT lymphoma. None of these was seen in inflammatory infiltrates. The presence of germinal centers, acute inflammation and reactive epithelial atypia does not exclude a diagnosis of low-grade gastric lymphoma. PMID- 9747123 TI - [Current developments in radiologic diagnosis of pancreatic diseases]. PMID- 9747122 TI - Calcifying odontogenic cyst: report of two cases and review of literature. AB - The "Histological Typing of Odontogenic Tumours" (W.H.O., 1992) classified the Calcifying Odontogenic Cyst (C.O.C.) into two variants: the non-neoplastic cystic C.O.C. and the odontogenic ghost cell tumour, which is predominantly solid. We reported two cases of C.O.C.: a case with intraosseous development and another with extraosseous localisation, in the soft tissue of the alveolar area. The first case represents a cyst delimited by a squamous, non-keratinized epithelium, thickened in some areas through the accumulation of ghost cells (big pale staining cells with a non-staining nuclear area). The connective tissue wall contains small ameloblastoma like islands. Dysplastic dentine islands, adjacente to the basal layer of the epithelium or in the connective tissue wall were also observed. The second case was a well-delimitated tumour consisting of ameloblastoma-like islands with numerous ghost cells inside. Islands of dysplastic dentine with psammomathous calcifications also exist. In certain histological sections microcystic aspects surrounded by ghost cells, dentinoid and ameloblastoma-like structures were noticed. The histochemical reaction for keratin and the immunohistochemical reaction for epithelial membrane antigen and for citokeratin were positive for ghost cells, suggesting their epithelial origin. Through this article we try to render pathologists sensitive with a particular and rare maxillary tumour. PMID- 9747124 TI - Experiences with gadodiamide, a non-ionic contrast agent, in MRI of brain metastases. AB - 29 patients suffering from brain metastases were studied with a single (0.1 mmol/kg body weight) and triple dose (0.3 mmol/kg) of gadodiamide, a non-ionic MR contrast agent. The study was performed using an extended imaging protocol with application of T1-weighted images in three orientations. Number of definite and suspected metastases, lesion contrast, edema delineation, and flow artefacts were evaluated. Most of the definite metastases were found in triple dose images. However, the total lesion number could only be determined by looking on all sequences as a whole. Furthermore, the number of suspected metastases could be significantly reduced with triple dose images, but once more all sequences were needed to do so. Flow artefacts were significantly increased by triple dose, whereby lesion contrast and edema delineation was considerably improved. The conclusion is that the combination of high dose application of the contrast agent and an optimized imaging protocol facilitates sufficient imaging of brain metastases. This is important for therapeutical considerations, because patients with more than 1-2 metastases will be treated primarily with radiation therapy instead of neurosurgery. PMID- 9747125 TI - [Embolization of the bronchial artery in recurrent hemoptysis]. PMID- 9747126 TI - [A new parameter for the determination of dose distribution in radiotherapy]. PMID- 9747127 TI - [Radiodiagnosis of elbow and shoulder--questions by the clinician]. PMID- 9747128 TI - [What is your diagnosis? Papillary fibroelastoma of the right coronary aortic sinus--embolism with 2 transient cerebral ischemia attacks]. PMID- 9747129 TI - [Reaction of the vascular wall to mechanical damage]. PMID- 9747130 TI - [Transanal endoscopic microsurgery--experiences at the Zurich University Hospital]. AB - The advantages of TEM (transanal endoscoic microsurgery) are minimally invasive, exact and full thickness excision of tumors in the rectum and a very low morbidity with excellent comfort for the patient. In a retrospective study all transanal endoscopic operations at Zurich University hospital in the last 5 years have been analyzed (n = 18). 11 adenomas and 5 carcinomas of the rectum have been resected with TEM (one mucosectomy, 16 full wall resections and one segmental resection of the rectum). In the group of the carcinomas there were four preoperatively known carcinomas, one T1 carcinoma was discovered postoperatively in the analyzed tissue. Among the four known carcinomas was one T1 carcinoma, two T2 carcinomas (one of them was thought to be a T1 preoperatively) and one T3 carcinoma. One patient with T2 carcinoma wanted specifically a minimally invasive procedure, the other one with T2 carcinoma was an older patient who didn't qualify for laparotomy. The patient with T3 carcinoma also had a malignant lymphoma. The operation was tolerated well by all the patients. There was one case of peritoneal perforation treated laparscopically and one case of postoperative bleeding. An incontinence of gas in one patient disappeared after 3 months. There was no adjuvant treatment in the group of the T1 carcinomas. One patient with a postoperative T2 carcinoma did not want a chemotherapy. The other two patients with T2 and T3 were polymorbid. Among the resected adenomas there was no case of recurrence. One T2 carcinoma recurred. These results show that transanal endoscopic microsurgery (TEM) is an excellent technique to treat ademomas and T1 carcinomas of the rectum with the advantages of full thickness excision under good vision, a minimal rate of recurrence and maximal patient comfort. The indications for transanal microsurgery are rare. The techically demanding operation is not always simple and should be performed in larger centers only. PMID- 9747131 TI - [Pre-eclampsia and its psychosocial sequelae]. AB - In the present work some psychosomatic conditions in the setting of preeclampsia are described. The important psychosocial consequences for women suffering from this disease and the drawback for their partners will be elucidated. Preeclampsia as a disease including hypertension, proteinuria and generalized edema is often associted with generalized seizures occuring most commonly at the end of the second trimenon of pregnancy. The disease bears a heavy risk for the mothers as well as for her unborn child. Until now the exact pathophysiological basis of the disease has not been entirely elucidated. For the pregnant woman and her psychosocial surrounding the outbreak of the disease is in most cases unexpected. During development of the disease she has to face a role change from a so far normal pregnancy to a high-risk situation. This may change also the attitude to the unborn child by herself and her partner. The preterm delivery induced therapeutically, together with the succeeding problems for the newborn complete the high psychosocial stress related to the entire situation. Therefore it is useful and important to offer psychosocial support to the mother as well as to her parter during the illness and the time after delivery. PMID- 9747132 TI - [Systemic corticosteroid therapy in rheumatology, advantages and risks]. AB - Apart from the therapy of autoimmune diseases, corticosteroids have an important position in the treatment of rheumatoid arthritis. Corticosteroids are used after the failure of non-steroidal antiinflammatory agents or of the basis therapies to control the illness. When the rheumatoid arthritis is accompanied by a systemic disease, they will be used earlier and in higher dosages. For polymyalgia rheumatica, independently of an association with temporal arteritis, corticosteroids are the therapy of choice. Risks of long-time corticosteroid therapy are a higher incidence of infection and bone demineralisation, especially in postmenopausal women. A careful prevention with Calcium and Vitamin D must be carried out systematically. The demineralisation can be limited by the use of Deflazacort, a corticosteroid, which decreases the loss of calcium. PMID- 9747134 TI - [Case from general practice. Agranulocytosis]. PMID- 9747135 TI - [Case from general practice. Bradycardic atrial flutter]. PMID- 9747136 TI - Blue Cross Blue Shield responds. PMID- 9747138 TI - Texas students take initiative. PMID- 9747137 TI - Advice about cognitive dysfunction. PMID- 9747139 TI - MedBytes. PMID- 9747140 TI - Forum on ethics. A lesson in informed consent. PMID- 9747141 TI - A conversation with the governor. Interview by Carlos Hamilton Jr and John P. Howe 3rd. PMID- 9747142 TI - The world of medicine in Texas. A look at international medical school graduates. PMID- 9747143 TI - The ultimate litmus test. PMID- 9747144 TI - Millennium meltdown. PMID- 9747145 TI - Statewide attitudes and behavior on child abuse and neglect in Texas. PMID- 9747146 TI - Gastric ulcer presenting as gastroesophageal reflux and apnea in a term neonate. AB - Apnea in the neonatal period frequently is associated with prematurity. Full-term infants who develop apnea usually have associated clinical conditions such as infection, shock, metabolic disorders, neonatal abstinence syndrome, intracranial pathology, and gastroesophageal reflux. Gastric ulcer also is a rare phenomenon in the neonatal period. We describe a full-term infant presenting with apnea. Upon investigation, a 6-channel pneumocardiogram revealed central apnea and multiple episodes of low esophageal pH (< 4), which is suggestive of gastroesophageal reflux. This was confirmed by an upper gastrointestinal series. A small antral ulcer crater also was demonstrated. When assessing the etiology of apnea in a full-term infant, gastroesophageal reflux and gastric ulcer should be considered. PMID- 9747147 TI - Calciphylaxis: a syndrome of skin necrosis and acral gangrene in chronic renal failure. AB - BACKGROUND: Calciphylaxis is a rare condition of rapidly extending ischemic skin necrosis or acral gangrene of fingers, toes or penis in patients with chronic renal failure. It may be accompanied by extensive metastatic calcification of soft tissues. Histology of infarcted tissues shows prominent medial calcification and intimal hyperplasia of subcutaneous arteries and/or digital arteries, respectively. The pathogenesis of calciphylaxis is only poorly understood. Most patients have hyperparathyroidism and an elevated calcium-phosphate-product, which is thought to be a major pathogenetic factor of calciphylaxis. PATIENTS AND METHODS: All published cases of calciphylaxis including nine of own (155 patients in total) from 1936 through 1996 were reviewed and subjected to statistical meta analysis (Fisher's exact test). RESULTS: Proximal locations of necrosis (thighs, buttocks, trunk) carried an unfavourable prognosis (63% mortality) compared to distal locations (calves, forearms, fingers, toes, penis) with 23% mortality (p < 0.0001). Parathyroidectomy was associated with a favourable outcome (p < 0.004). Diabetics with chronic renal failure had acral gangrene in 61% compared to 34% of the non-diabetic calciphylaxis-patients (p < 0.007). CONCLUSIONS: The present analysis of all published cases of calciphylaxis is limited by patient selection and publication bias. The unfavourable prognosis of patients with proximal necrosis is impressive and might justify an early and aggressive treatment in such cases. However, the general benefit of parathyroidectomy remains debatable. Hyperparathyroidism should be managed primarily by conservative means. Parathyroidectomy should be reserved for patients with very high parathyroid hormone level and calcium-phosphate-product or with a rapidly progressive disease. PMID- 9747148 TI - Localisation of protein Z in vascular lesions of patients with atherosclerosis. AB - BACKGROUND: The deposition of protein Z was investigated in atherosclerotic vascular lesions of patients with diabetes mellitus or atherosclerotic vascular disease without diabetes in comparison to controls. PATIENTS AND METHODS: Protein Z antigen was evidenced by immunohistochemistry in arteries of 5 healthy control patients, 11 diabetic patients with arterio-occlusive disease and 7 patients suffering from arterio-occlusive disease without diabetes. For immunohistochemistry, a commercially available antibody was taken as first antibody, and immunopositivity was evaluated independently by two investigators (J.G.; I.K.) as negative (0), positive (+) and strongly positive (++). The results were assessed by the Whitney-Mann-Wilcoxon test. RESULTS: Macrovascular endothelial cells were stained positive for protein Z in all arteries studied. Arteries of controls did not show significant immunopositivity in cells other than macrovascular endothelial cells, while the microvascular endothelial cells of control arteries were largely negative. The proliferating subendothelial space in atherosclerotic vascular lesions showed significant immunopositivity for protein Z. In contrast to control arteries, the microvascular endothelial cells of the proliferating areas stained positive. The staining pattern of the subendothelial space was similar in atherosclerotic vessels independent of the risk factor for atherosclerosis. Plaques were immunopositive for protein Z, too. CONCLUSIONS: Protein Z is present in atherosclerotic vascular lesions of diabetic and non-diabetic patients, but not in the subendothelial space and microvascular endothelial cells of healthy controls. Since protein Z-positivity was detected in microvascular endothelium as well as in extra-vascular deposits around plaques, it may play a role in the development of these lesions. PMID- 9747150 TI - Discrepancy of clinical and angiographic results in the follow-up of percutaneous transluminal renal angioplasty (PTRA). AB - BACKGROUND: To investigate the value of recurrent hypertension as indicator for renal artery patency after PTRA clinical and angiographic follow-up results were analysed. PATIENTS AND METHODS: results of 66 follow-up angiographies and blood pressure measurements were available in 55 patients after technically and clinically successful PTRA. Out of 43 patients with atherosclerosis, in 33 was recurrent hypertension present, and in ten patients none. Of 12 angiographies in patients with fibromuscular dysplasia nine were done because of recurrent hypertension. RESULTS: In atherosclerosis, 21 (64%) out of 33 patients with recurrent hypertension showed a re-stenosis by > 70%, and 12 (36%) none. Out of the ten follow-up angiographies performed in patients with no recurrence, two showed an unexpected stenosis. In fibromuscular dysplasia, stenoses were present in seven cases (77%). Four of them were at the site of PTRA (44%), while three were at a new site (33%). The follow-up angiographies in three patients with no recurrence showed patent arteries. Out of 11 further angiographies carried out because of hypertension after repeated PTRA, only three revealed re-stenosis. CONCLUSIONS: In only about 35% of all patients with recurrent hypertension re stenosis was shown in angiography but in 15% of asymptomatic patients. Recurrence of hypertension after PTRA and renal artery stenosis is not well correlated. Thus, follow-up should be performed not only clinically but also by direct examination of renal artery patency such as by ultrasound. PMID- 9747149 TI - Angiotensin-converting enzyme gene insertion/deletion polymorphism and peripheral arterial occlusive disease. AB - BACKGROUND: The deletion polymorphism of the ACE gene is linked to a high risk of cardiovascular disease due to the permanent activation of the local and systemic renin-angiotensin systems (RAS). The aim of this prospective study was 1. to compare the ACE insertion/deletion polymorphism in individuals with a healthy vasculature with that of patients suffering from peripheral arterial occlusive disease (PAOD), and 2. to determine whether associations existed between specific clinical parameters and the ACE genotype which the PAOD patients expressed. PATIENTS AND METHODS: Determinations of ACE I/D gene polymorphism were made using a polymerase chain reaction (PCR) technique on 98 patients with clinical stage II PAOD according to Fontaine and 240 healthy individuals who served as controls. All patients and controls came from central Germany. Clinical variables which included duration of clinical symptoms, a familial history of the disease, arteriosclerosis score (ASF, providing an estimate of the extent of atheromatosis at femoral artery bifurcation) and plasma ACE activity were correlated with the genotypes taking the cardiovascular disease risk factors which were present into consideration. RESULTS: Differences in ACE genotypes between patients with PAOD (D/I: 0.57/0.43) and control group individuals (D/I: 0.59/0.41) were not observed. In comparison with the II genotype, the DD genotype was associated with a shorter duration of disease (p = 0.01), a positive family medical history (p = 0.022) and a higher plasma ACE activity (p = 0.026). The ASF did not correlate with the ACE I/D gene polymorphism. CONCLUSION: Evidence that the deletion allele is linked to the manifestation of PAOD could not be found in the patients studied. One can assume, however, that the deletion allele has a progression promoting effect on the disease. PMID- 9747151 TI - Does the measurement of digital arterial pressure using the Doppler ultrasonic technique without testing for vasospasm to detect digital arterial occlusions make good sense? AB - BACKGROUND: An attempt was made to determine whether measurement of the systolic pressure with the Doppler ultrasonic technique can be used to detect occlusions of the digital arteries. PATIENTS AND METHODS: A total of 92 patients (44 women, 48 men) with an average age of 47.5 years (range: 19-82 years) were investigated. Of these, 17 were diagnosed with a primary Raynaud's syndrome, 45 with a secondary, and 5 patients with a suspected secondary Raynaud's syndrome; 25 patients had digital occlusions of varying genesis. In all patients, the Doppler ultrasonic technique was applied to measure the systolic pressure of the digital arteries of both hands at a room temperature of 21-24 degrees Celsius without any additional thermal (warm or cold) provocation or use of vasodilatory agents; the results obtained were compared with arteriographs of the hands. RESULTS: While the sensitivity of the Doppler ultrasonic technique was a very low 55%, the specificity was a very high 95%. With a prevalence of 68%, the positive predictive value was 96%, the negative predictive value 50%. In the case of angiographically patent digital arteries with no spastic narrowing, the Doppler ultrasonic-measured systolic pressure was up to 30 mmHg above, and up to 25 mmHg below, the systolic pressure measured in the ipsilateral brachial artery. A pressure gradient between upper arm and digital arteries of this order of magnitude therefore does not exclude pathological changes. Only angiography was able reliably to detect vascular stenoses and occlusions. CONCLUSION: Reliable exclusion of digital arterial occlusions using the Doppler ultrasonic technique without testing for vasospasm is not possible. Only occlusions of digital arteries together with simultaneous occlusions of hand arteries can be reliably detected. Owing to the considerable scatter of the systolic blood pressure gradient between brachial and digital arteries, the Doppler ultrasonic technique cannot be used to distinguish between patent and occluded digital arteries without the use of additional vasodilation. PMID- 9747152 TI - Influence of changes in arterial blood pressure and peripheral arterial resistance on peak systolic velocity ratio. AB - BACKGROUND: Peak systolic velocity ratio has been described as a parameter to determine the degree of arterial stenosis. But there is very little information about the influence of changes in arterial blood pressure or peripheral arterial resistance during exercise on the peak systolic velocity ratio. PATIENTS AND METHODS: Peak systolic velocity was calculated before and in arterial stenosis in 35 patients with only single stenosis in the femoral or iliacal arteries under 4 different conditions: a) twice under resting conditions as a control, b) increased blood pressure by arm activation but unchanged peripheral vascular resistance, c) increased blood pressure by leg activation with a reduced peripheral vascular resistance by metabolic vasodilatation, d) decreased blood pressure associated with pharmacologically reduced peripheral resistance (10 mg nifedipine). RESULTS: Peak systolic velocity ratio was: a) 5.8 +/- 3.7 and 5.7 +/ 3.3, b) 5.6 +/- 3.6 (the increase in systolic arterial blood pressure was 20 +/- 3 mmHg), c) 6.3 +/- 4.4 (increase in systolic arterial blood pressure was 21 +/- 3 mmHg), d) 5.6 +/- 3.4 (decrease in systolic arterial blood pressure was 18 +/- 8 mmHg) without being significantly different from each other. The correlation factors of the peak systolic velocity ratios to the angiographic diameter reduction were between 0.737 and 0.847. Although the mean values suggest that there is no influence from the different exercise tests or nifedipine application on the peak systolic velocity ratio single stenosis demonstrated large reproducible differences. CONCLUSION: The influence of changes in arterial blood pressure and peripheral resistance on peak systolic velocity ratio appeared small. But a single stenosis showed large increases or decreases of peak systolic velocity ratio possibly due to vasomotion of the prestenotic or stenotic arterial segment. PMID- 9747153 TI - Comparative results of thrombolysis treatment with rt-PA and urokinase: a pilot study. AB - BACKGROUND: The aim of the following prospective study was to investigate whether patients benefited from locoregional lysis treatment of recent deep leg vein thrombosis after 1 year. PATIENTS AND METHODS: The prospective study included 69 patients aged between 22 and 58 years, in whom recent lower leg vein and popliteal vein thromboses were diagnosed by phlebography. Patients were randomized to one of three treatment groups: one group was treated for a maximum of 7 days with full heparinization and daily dose of 20 mg rt-PA administered locoregionally over a period of 4 hours; a second group received 100,000 IU/h urokinase locoregionally for a maximum of 7 days, in addition to full heparinization; and in the third group (control group), intravenous heparin infusions after PTT constituted the only form of treatment. All patients were given phenprocoumon from day 7 and received compression treatment. Before treatment began and before the course of phenprocoumon started, phlebography and colour duplex sonography examinations were carried out. After 12 months, follow up duplex sonography was conducted to evaluate the reflux times over the popliteal vein and the degree of patency of the deep leg veins. RESULTS: Complete lysis was achieved in 6 of 22 patients in the recombinant tissue plasminogen activator (rt-PA) group and in 11 of 22 patients in the urokinase group. At follow-up examination after 12 months, there were serious post-thrombotic changes in 14 of 22 patients in the rt-PA group, in 9 of 22 patients in the urokinase group and in 15 of 22 patients in the group of patients who received no lysis treatment. CONCLUSION: Patients with recently formed thromboses in the lower leg and popliteal veins who underwent 7 days of locoregional lysis treatment with urokinase demonstrated significantly fewer clinical symptoms of post-thrombotic syndrome after 1 year than those who received locoregional treatment with rt-PA over a similar period or a control group treated with anticoagulants only. PMID- 9747154 TI - Thrombosis of the muscular calf veins--reference to a syndrome which receives little attention. AB - BACKGROUND: Thrombosis of the soleal and gastrocnemial veins seldom receive very much attention. It is thought that they are frequently the starting point for deep vein thrombosis of the lower extremity. PATIENTS AND METHODS: 125 patients with clinically suspected deep vein thrombosis of the leg were examined by means of duplex sonography and phlebography. The ultrasound examination was performed on the lower limb in the form of a compression sonography. RESULTS: From 137 legs examined, a total of 82 cases of deep vein thrombosis were diagnosed. Sonographically and/or phlebographically, the soleal and/or gastrocnemial veins were found to be involved in 65 cases of thrombosis, i.e. 79% of all thrombosis. 25% of all cases of deep vein thrombosis of the leg were isolated muscular vein thrombosis. Pain in the calf while walking, similar to muscular soreness after exertion, was typical of the isolated muscular vein thrombosis. The muscular veins were involved in all the deep vein thrombosis of the leg of the 3- and 4 layer type. Diagnosis of soleal and gastrocnemial vein thrombosis is quite possible by means of sonography and phlebography. The sensitivity and specificity of the compression sonography were 88% and 95% respectively, compared to the phlebography. CONCLUSION: In patients suffering from pain in the calf, the soleal and gastrocnemial veins should be carefully included in the sonographic or phlebographic assessment. Due to the risk of deep vein thrombosis, isolated muscular vein thrombosis should receive treatment appropriate for a deep vein thrombosis of the calf, and its development be checked. PMID- 9747155 TI - Hand-arm vibration syndrome with proximal ulnar artery occlusion. AB - The case of a man exposed during 25 years to vibration while maneuvering a heavy earth moving tractor is reported. The first clinical manifestation of hand-arm vibration syndrome was a bilateral Raynaud's phenomenon followed five years later by digital necrosis. The arteriography revealed a proximal and bilateral ulnar artery occlusion. Bilateral median nerve conduction abnormalities were also present. Vibration exposure level was much higher than the threshold level proposed by the European Commission. Laboratory examinations for vasculitis and other vascular diseases were all negative. The purpose of this report is to show that vibration can be responsible for proximal occlusion of a medium sized artery and severe neurovascular abnormalities which must be distinguished from the usual vasospastic Raynaud's phenomenon and the classical hypothenar hammer syndrome. An early and correct diagnosis is crucial because continued repetitive trauma can result in irreversible ischemic injury and loss of digits. PMID- 9747156 TI - Cystic adventitial degeneration and entrapment syndrome of the popliteal artery as a differential diagnosis of popliteal stenosis or occlusion in the younger age group. AB - Differential diagnosis in angiographically found popliteal artery stenosis or occlusion comprises some distinct and clinically important entities that should be considered as management and prognosis may vary considerably. We present two patients with the final diagnosis of cystic adventitial disease of the popliteal artery and popliteal artery entrapment syndrome. Angiographic findings and the value of additional diagnostic imaging are discussed. PMID- 9747157 TI - [Arteriovenous fistula of the jugular vein: detection and follow-up with color coded duplex ultrasound]. AB - A 71-year-old man presented with a pulsating swelling in the lower right neck region. On clinical examination palpation revealed a local thrill and auscultation the typical systolic vascular sounds. On sonography the typical findings for an AV fistula with large widening of the jugular vein and entry of an extremely rapid jet stream into the vein were detected. The entry point was under a venous valve whose movements apparently influenced the jet stream since the inflow was only seen intermittently and only in the systolic phase. The shunt volume of the fistula was apparently small: indirect signs of an arteriovenous short circuit could not be seen on duplex sonography. The low-flow fistula was not demonstrated by arterial DSA. Thus, the arterial sources could not be identified. Sonographic monitoring over 5 years at first showed no changes while at the last follow-up the fistula was no longer detectable. The suspected cause of the arteriovenous fistula was a vascular wall perforation effected some years previously during insertion of a central venous catheter. We further assume that the anticoagulation therapy with Marcumar started at that time had prevented spontaneous closure of the fistula over a period of several years. This case illustrates the value of colour-coded duplex sonography for the differentiation of pulsating tumors in the neck. In individual cases of AV fistulas with low shunt volumes the method is apparently superior even to angiography. PMID- 9747158 TI - [Pelvic and leg vein thrombosis in azygous and hemi-azygous vein continuity syndrome and complete agenesis of the inferior vena cava]. AB - A patient with a complete agenesis of the inferior vena cava is presented. In this rare anomaly the blood of the pelvic veins is drained through the dilated venae lumbales, vena azygos and vena hemiazygos into the superior vena cava. The malformation was detected within the scope diagnostics of deep vein thrombosis. The diagnosis was made by sonography, computed tomography and magnetic resonance angiography. PMID- 9747159 TI - [Hereditary benign telangiectasia: a rare form of primary telangiectasia with successful treatment with flash-lamp-pumped pulsed dye laser]. AB - We report about multiple cutaneous telangiectases occurring over 3 generations. The pattern of inheritance, the morphology of the lesions, and the absence of hemorrhagic episodes are consistent with an unusual variant of primary telangiectasia described as hereditary benign telangiectasia. Differential diagnosis and pathogenesis of this rare type of telangiectasia is discussed. The lesions have been treated with the flashlamp-pumped pulsed dye laser. The lesions were completely removed by two treatments. Six months later, the patient had no recurrence of the telangiectases. Without producing scars or permanent pigmentary changes a very good cosmetic result has been achieved and stigmatisation and psychic burden of this patient has been relieved. PMID- 9747160 TI - Excising the reexcision: stereotactic core-needle biopsy decreases need for reexcision of breast cancer. AB - There is debate regarding use of the stereotactic core-needle biopsy (SCNB) for highly suspicious mammographic lesions. This study compares a serial group of mammography-detected breast cancer patients treated before and after the use of SCNB. We studied 113 consecutive nonpalpable breast cancers between 1994 and 1996. Altogether 47 patients were diagnosed by wire-localized breast biopsy (wire group) and the next 66 consecutive breast cancer patients by SCNB (stereo group). Negative margins were found more often in the stereo group than in the wire group (77% vs. 38%, p < 0.001). Reexcision was required more frequently in the wire group than in the stereo group (68% vs. 21%, p < 0.001), and one-staged surgical procedures were done more often in the stereo group than the wire group (79% vs. 21%, p < 0.001). The volume of the initial wide excision was much larger in the stereo group than in the wire group (p = 0.002). Those in the wire group required 50% more operations per patient (1.8 vs. 1.2) than the stereo group. A significant cost savings can be estimated in the stereo group compared with the wire group. The use of SCNB was associated with breast excisions of larger volume, negative margins, and decreased need for reexcision. Simultaneous adjunct procedures resulted in one-stage operations, improving cost savings. The use of SCNB for nonpalpable breast cancer benefits the patient, the surgeon, and the payor. It should be undertaken prior to the first surgical procedure. PMID- 9747161 TI - Short-term outcome and predictors of adverse events following pulmonary thromboendarterectomy. AB - Pulmonary complications including hypoxemia, right heart failure, and prolonged ventilation may follow pulmonary thromboendarterectomy (PTE) performed via cardiopulmonary bypass (CPB) with deep hypothermic circulatory arrest. Seventeen adult patients have undergone PTE at the University of Maryland Medical System during the preceding 3 years. From these patients, clinical and hemodynamic parameters were tabulated pre-CPB, post-CPB, at admission to the intensive care unit (ICU), and prior to discontinuation of invasive monitoring in the ICU. Data on anthropometric variables, survival, and times of extracorporeal circulation, mechanical ventilation, and hospital stay were also collected. The mean values for pulmonary arterial systolic and diastolic pressures and pulmonary vascular resistance (PVR) decreased significantly from pre-CPB values after PTE (all p < 0.05). Mild mixed acidosis present at ICU admission resolved prior to discharge (p = 0.002). The length of mechanical ventilation time was positively correlated with the absolute post-CPB PVR and negatively correlated with the relative change in central venous pressure (CVP) from pre-CPB to post-CPB values (r = 0.75, p = 0.037). Of the pre-CPB anthropometric variables, only body mass index was significantly higher in nonsurvivors (p = 0.037). Pulmonary artery pressures and vascular resistance fall significantly after PTE. A lower post-CPB PVR and a relatively decreased (i.e., from pre-CPB values) CVP predict reduced length of postoperative ventilation but not of the hospital stay. Mortality appears increased in patients with a large body habitus. PMID- 9747162 TI - Transpelvic gunshot wounds: routine laparotomy or selective management? AB - Mandatory exploration is the standard method for managing patients with gunshot wounds to the abdomen and back. This policy is associated with a high incidence of unnecessary laparotomies and significant morbidity. Reports from our center have shown that a policy of selective management, based on clinical findings, is safe in such patients. Patients with bullet trajectories that carry a high likelihood for intraabdominal organ injury may constitute a subgroup at particular risk. The need for routine or selective exploration in similar patients must be assessed. Therefore we decided to analyze patients with transpelvic gunshot wounds. The objective of the study was to examine if a policy of selective management of patients with transpelvic gunshot wounds is safe. This prospective study was conducted at an academic level I trauma center. We admitted 37 patients with transpelvic gunshot wounds over a 12-month period. All patients were managed according to a protocol that dictated laparotomy in the presence of significant clinical findings (peritoneal signs, hemodynamic instability, gross hematuria, rectal bleeding) and observation in the absence of the above. Additional diagnostic workup was performed only in appropriate cases rather than routinely. Nineteen (51.3%) patients were immediately operated on the basis of clinical findings. Sixteen of these laparotomies were therapeutic. Eighteen (48.6%) patients were initially observed. Subsequently, three of them underwent exploration for development of abdominal tenderness. All three laparotomies were nontherapeutic. The remaining 15 (40.5%) patients were successfully managed nonoperatively. There were no delays in diagnosis or missed injuries. Clinical examination had a sensitivity of 100% and specificity of 71.4% in detecting the need for laparotomy. A policy of selective management is thus safe, even for patients who suffer gunshot wounds with a high likelihood for intraabdominal organ injury. Clinical examination, supported by additional studies in appropriate cases, is the main method of selecting patients for operation or nonoperative treatment. PMID- 9747163 TI - Arterial reconstruction: justified for patients with intermittent claudication? AB - The objective of this study was to evaluate the effects of arterial reconstruction in patients with intermittent claudication. A total of 243 patients (305 limbs) underwent lower extremity vascular reconstruction at our institution from 1979 to 1995. They were assessed by physical examination, pulse volume recordings, segmental pressure, Duplex ultrasonography, and intravenous subtraction arteriography to evaluate the effects of arterial reconstruction. Surviving patients (220 limbs) were enrolled for evaluation of outcome during the follow-up period. There were 59 deaths during the follow-up period. The cumulative life-table 5-year patency rates were 90% +/- 3%, 73% +/- 6%, and 74% +/- 10% for aortoiliac, infrainguinal, and aortofemorodistal arterial reconstructions, respectively. Among 129 repairs in the aortoiliac region, Fontaine stages I, II, and III were found in 109 limbs (84.5%), 17 limbs (13.2%), and 2 limbs (1.5%), respectively. There was one (0.7%) minor amputation. There was Fontaine stage I in 50 limbs (76.9%), Fontaine stage II in 14 limbs (21.5%), and Fontaine stage III in 1 limb (1.5%) for repairs in the infrainguinal region. Among the 26 aortofemorodistal repairs, there was Fontaine stage I in 21 limbs (81%) and Fontaine stage II in 5 limbs (19%). There was a statistically significantly higher incidence of Fontaine stage I than Fontaine stage II or III in aortofemoral, infrainguinal, and aortofemorodistal arterial reconstructions (p < 0.0001). Arterial reconstruction for patients at Fontaine stage II offered benefits and improved quality of life at follow-up. Arterial reconstruction for patients at Fontaine stage II offers benefits and improved quality of life at follow-up. It was concluded that the significant improvement in quality of life after arterial reconstruction warrants continued use of the procedure in patients with intermittent claudication. PMID- 9747164 TI - Manometric findings of the upper esophageal sphincter in esophageal achalasia. AB - Pharyngeal and upper esophageal sphincter (UES) manometry was performed in 15 patients with esophageal achalasia and compared with that in 10 healthy controls. Neither the pharyngeal contraction pressure nor the UES resting pressure were significantly different between the two groups, although the UES residual pressure in patients with achalasia was significantly increased compared with that in controls. Pneumatic dilatation of the lower esophageal sphincter (LES) was performed in these patients. After successful LES dilatation, the increased UES residual pressure in patients with esophageal achalasia decreased significantly. Our results suggest that UES relaxation in patients with esophageal achalasia is incomplete compared with that in healthy adults. This UES abnormality is not a primary defect but a secondary phenomenon. PMID- 9747166 TI - Carcinomatous infiltration into the submucosa as a predictor of lymph node involvement in early gastric cancer. AB - The clinicopathologic features of 114 patients with resectable early gastric cancer (EGC) invading the submucosa were examined retrospectively with respect to lymph node involvement and the possibility of performing a minimally invasive operation. Patients were divided into node-positive (n = 25) and node-negative (n = 81) groups. Among several pathologic factors, the diameter of the tumor and lymphatic involvement were significantly correlated with nodal involvement. Within the submucosal layer the depth of invasion and the horizontal cancerous expansion also correlated with lymph node disease (p < 0.05). The size of the tumor did not correlate with the length of submucosal infiltration (r = 0.12, p = 0.1). Patients with both slight invasion into the submucosa and less than 5 mm of horizontal expansion were often negative for lymph node involvement and thus may benefit from local surgery as an alternative to gastrectomy. PMID- 9747165 TI - Total or subtotal gastrectomy for gastric carcinoma? A study of quality of life. AB - The aim of this study was to compare quality of life after total gastrectomy (TG) with that after subtotal gastrectomy (STG) for gastric carcinoma. The value of the routine use of TG de principe in the treatment of gastric carcinoma, wherever the tumor may be sited in the stomach, remains controversial. The advocates of TG contend that when it can be performed safely, with relatively low operative mortality and morbidity, it yields better long-term survival than STG. Most surgeons, however, believe that the routine use of TG increases both operative mortality and morbidity and the risk of nutritional deficiency in the long term, without improving survival. TG may also be associated with poorer outcome in terms of quality of life (QOL), but the evidence for this is tenuous. Forty-seven consecutive patients who had undergone potentially curative (R0) gastric resection for carcinoma were studied: 26 had undergone TG and 21 STG. A radical D2 lymph node dissection had been performed in each, and all patients were free from recurrence at the time of the study. QOL was measured before operation and 1, 3, 6, and 12 months after operation by means of five questionnaires to measure functional outcome: the Rotterdam symptom checklist (RSCL), the Troidl index, the hospital anxiety and depression (HAD) scale, activities of daily living score, and Visick grades. Before operation there was no significant difference in QOL between the two groups of patients. At 1 year after operation, however, patients who had undergone STG had a significantly better QOL than patients who had undergone TG: Their median RSCL score was lower (10 versus 19 respectively, p < 0.05), and their Troidl index was higher (11 versus 9 respectively, p < 0.05). The QOL of patients who underwent STG was also significantly better after operation than it had been before operation, whereas the QOL of the TG group was not significantly better after operation than before operation. The QOL of patients was found to be significantly better after STG than after TG for gastric carcinoma. Because operative mortality is greater and long-term survival is no better after TG than after STG, the latter is recommended as the treatment of choice for tumors of the distal stomach. PMID- 9747167 TI - Effect of duodenal distension on the pyloric sphincter and antrum and the gastric corpus: duodenopyloric reflex. AB - The mechanical effect of balloon distension of the duodenum on the stomach was studied in 10 mongrel dogs with a mean weight of 14.8 +/- 3.2 kg. The response of the pyloric sphincter and antrum as well as of the corpus of the stomach to duodenal distension by a balloon filled with water in increments of 2 ml, up to 6 ml, was determined. The test was repeated after anesthetizing the pyloric sphincter and antrum and the duodenum, each at a separate time. In 5 of 10 dogs the effect of duodenal distension on the vagotomized stomach was studied. Duodenal distension with 2 ml of water produced an increase in the pyloric sphincter pressure (p < 0.05) and a decrease in the antral pressure (p < 0.05); it had no effect on corporeal pressure (p > 0.05). Distension with 4 ml and 6 ml produced the same effect as 2 ml (p > 0.05). The anesthetized pyloric sphincter and antrum did not respond to duodenal distension. Likewise, the pyloric sphincter and antrum showed no response to distension of the anesthetized duodenum or of the duodenum after vagotomy. Pyloric sphincter contraction and antral dilatation upon duodenal distension suggest a reflex relation we call the duodenopyloric reflex. This reflex appears to prevent duodenopyloric reflux. Moreover, the antrum dilates probably to accommodate more gastric contents. PMID- 9747168 TI - Perforation due to metastatic tumors of the ileocecal region. AB - We reviewed our department's medical records between April 1986 and April 1994 to identify patients who showed acute abdominal symptoms requiring surgical treatment due to metastatic tumors of the small intestine. In group A, seven patients (30%) were treated for acute peritonitis, and all were found to have an intestinal perforation due to hematogenous metastases (group A). In group B, 16 patients (70%) were treated for an intestinal obstruction, and all were found to have disseminated tumors of the small intestine (group B). In group A all tumors were isolated and located exclusively in the ileocecal region, whereas all tumors in group B showed peritoneal dissemination, with no predominant anatomic localization. In general, the intestinal tumors in group A originated from cancers of the upper aerodigestive tract, whereas those in group B originated from advanced cancers in the abdominal cavity. The tumors were significantly smaller and the period between the onset of symptoms from the original malignancy and the onset of abdominal symptoms (perforation or obstruction) was significantly shorter in group A. In conclusion, intestinal metastases located in the ileocecal region have unique clinicopathologic features and so should be recognized as a distinct disease entity. Therefore when patients with a known upper aerodigestive malignancy exhibit acute abdominal symptoms, intestinal metastasis to the ileocecal region, necrotic changes, and perforation should be considered in the differential diagnosis. PMID- 9747169 TI - Wound healing of intestinal anastomosis after digestive surgery under septic conditions: participation of local interleukin-6 expression. AB - This study aimed to evaluate the integrity of anastomotic wound healing after digestive surgery under septic conditions and to observe local interleukin-6 (IL 6) expression around the anastomotic segment. Experimental animals were separated into lipopolysaccharide (LPS) and control groups. Each was injected with LPS or normal saline solution into the peritoneal cavity 24 hours before transection and anastomosis of the colon. The anastomotic bursting pressure (ABP) and tissue hydroxyproline concentration (HP) were measured as indicators of wound healing. Immunohistochemical staining for IL-6 was performed on tissue samples obtained from the anastomotic segment, lung, liver, and kidney. The reactive cells were counted by light microscopy. The ABP and HP were significantly lower in the LPS group than the control group 7 days after the surgery. In the LPS group, IL-6 expression around the anastomotic segment was enhanced 1 and 6 hours after surgery but suppressed 24 hours afterward. In contrast, IL-6 expression in lung, liver, and kidney was enhanced in the LPS group 24 hours after surgery but not in the control group. It is suggested that anastomotic wound healing is impaired after digestive tract surgery under septic conditions, and local IL-6 expression participates in wound healing. PMID- 9747170 TI - Increased serum interleukin-8: correlation with poor prognosis in patients with postoperative multiple organ failure. AB - This study investigated whether cytokines and colony-stimulating factors can predict prognosis in patients with postoperative multiple organ failure (MOF). We evaluated 14 patients with postoperative MOF who underwent operation for cardiovascular disease. Seven patients recovered from MOF (survivors) and seven did not recover and died (nonsurvivors). The white blood cell (WBC) count, granulocyte colony-stimulating factor, monocytic colony-stimulating factor, interleukin-6 (IL-6), and IL-8 were measured on the day the patients were judged to be in MOF and each week thereafter until the patients recovered or died. Survivors and nonsurvivors were equivalent in terms of age, gender, proportion of use of extracorporeal circulation, operation time, volume of blood transfusion, time from operation to the onset of MOF, the MOF score, proportion of bacteremia, duration of MOF, and number of failed organs. The mean duration of MOF was less than 2 weeks in both groups; therefore the measurements were compared on the first day of MOF and 1 week later. No significant differences between the two groups in terms of WBC counts, colony-stimulating factors, and IL-6 levels were noted. However, the serum level of IL-8 was significantly higher in nonsurvivors than in survivors. Patients with a high serum levels of IL-8 at the time of MOF had a poor prognosis. PMID- 9747171 TI - Portal vein thrombosis following splenectomy for hematologic disease: prospective study with Doppler color flow imaging. AB - We report the results of a prospective series of 60 consecutive splenectomies for hematologic disorders performed between February 1995 and May 1996. The portal venous flow of all the patients (34 men and 26 women with a mean age of 54.1 years) was systematically studied before and after intervention with Doppler color imaging (on the day before the intervention and on the 7th and 30th postoperative days). The objective of this study were to determine the real frequency of asymptomatic portal or splenic venous thrombosis (PSVT) after hematologic splenectomy. The intervention began with exteriorization of the spleen and the tail of the pancreas; ligation of the splenic vein was performed close to its junction with the inferior mesenteric vein. Twenty-three complications (38.3%) were noted with three deaths (5%). One symptomatic PSVT (1.6%) and three asymptomatic PSVTs (6.7%) were diagnosed and treated with no deaths. Three risk factors of PSVT, recognized by all the authors, were present in these four cases: large splenomegaly, thrombocytosis, or myeloproliferative disorder. The systematic ultrasonographic (US) examinations increased the frequency of diagnosis of PSVT sevenfold during the perioperative period. Patients with marked splenomegaly associated with lymphoma, chronic lymphocytic leukemia, or myeloid metaplasia probably require systematic US monitoring during follow-up, but this must be determined by further study. PMID- 9747172 TI - Repeat hepatectomy for recurrent colorectal metastases. AB - Recurrence rates after hepatic resection in patients with colorectal metastases are reported to range from 47% to 80%. Hepatic recurrence is seen in 35% to 50% of patients. Aggressive surgical resection appears to be a worthwhile treatment in patients with recurrent hepatic metastases to promote longer patient survival because surgical resection remains the only curative therapy available. This is a retrospective review of our experience with 15 patients undergoing repeat hepatic resection culled from 67 patients undergoing initial hepatectomy for metastatic colorectal cancer. Of 67 patients who underwent hepatectomy for colorectal hepatic metastases, 33 developed hepatic recurrence at a median interval of 23 months (range 1-176 months) after the first hepatectomy. The second hepatectomy was performed in 15 patients 5 to 29 months after the first hepatectomy, with no mortality. The mean operating time and blood loss at the second hepatectomy were similar to those at the first hepatectomy. The mean hospital stay at the second hepatectomy was significantly shorter than that at the first hepatectomy. The cumulative survival rate for the 15 patients was 42.4% at 3 years and 21.2% at 5 years, respectively, which compared favorably with the survival rate of the 67 patients who underwent initial hepatectomy. Patients who underwent the second hepatectomy had significantly higher survival rates from the first hepatectomy than the 18 patients with unresectable hepatic recurrence. Repeat hepatectomy can be performed safely and provides long-term survival rates similar to those of first hepatectomies. In appropriately selected patients, repeat hepatectomy for colorectal metastases is a worthwhile treatment. PMID- 9747173 TI - Efficacy of PdB in preventing intraoperative risk of infectious diseases. AB - The objectives of this study were to (1) determine the number of punctures surgeons and assistants suffer during operations involving a laparotomy during the intraabdominal and closure phases; and (2) determine if the number of puncture injuries during wound closure can be reduced using a new surgical instrument (PdB) that protects the surgeon's hands and the patient's viscera against needlesticks. For the first objective, all laparotomies performed during 1 month (n = 52) were controlled, collecting the gloves used and determining the number of perforations. For the second objective, a randomized prospective controlled study, involving two series of 100 medial laparotomies, was carried out. The incidence of perforations was 29% during the intraabdominal phase and 16% during the wound closure phase. The glove perforation rate while closing medial laparotomies was 31.5% if the PdB was not used and 3% if the PdB was used (p < 0.0001). The glove perforation rate during laparotomy is significant, but with the use of the PdB this incidence can be significantly reduced. PMID- 9747174 TI - Surgical significance of supernumerary parathyroid glands in renal hyperparathyroidism. AB - In secondary hyperparathyroidism (2HPT) fundamentally all parathyroid glands, including supernumerary glands, become hyperplastic, and stimulation of parathyroid glands continues after parathyroidectomy (PTx). Therefore supernumerary glands have special significance during surgery for 2HPT, whether persistent or recurrent HPT. In the present study 570 patients underwent initial total PTx with a forearm autograft. The frequency, type, location, histopathology, and clinical significance of the supernumerary glands were evaluated. At the initial operation 90 supernumerary glands were removed from 82 to 570 patients (14.4%); 12 patients (2.1%) required extirpation of supernumerary glands for persistent/recurrent HPT. Altogether 104 supernumerary glands were identified at operation in 94 of the 570 patients (16.5%). Among these 104 glands, 25 (24.0%) were of the rudimentary, or split, type and 79 (76.0%) of the proper type. Supernumerary glands were most frequently identified in the thymic tongue (53/104, 51.0%); 32 (60.4%) of these 53 glands were identified only microscopically. In 6 of the 570 cases (1.1%), reoperation was required for persistent HPT due to supernumerary glands located in the mediastinum, and 6 patients underwent neck reexploration for recurrence. Histopathologically, 61 of 104 (58.7%) supernumerary glands, including 36 glands recognized only microscopically, showed diffuse hyperplasia, and 43 (41.3%) displayed nodular hyperplasia. Residual small supernumerary glands with diffuse hyperplasia have the potential to be transformed to nodular hyperplasia during long-term hemodialysis. Therefore all parathyroid glands including supernumerary glands should, if possible, be removed at the initial operation. Routine removal of the thymic tongue and careful examination of the regions surrounding the lower poles of the thyroid, especially on the left side, are important steps in the surgical treatment. PMID- 9747175 TI - Historical evolution of hypothermic liver surgery. AB - The clinical application of hypothermia dates back to the surgical treatment of blue babies (1949) and the early days of open heart surgery (1952), when generalized cooling was employed. The induction of hepatic hypothermia began with whole-body cooling in experimental models in 1953 and clinically in 1961. It was designed to minimize the ischemia-reperfusion injury associated with hepatic inflow occlusion. Body surface cooling and cooling via an extracorporeal circuit, however, were not widely accepted for hepatic surgery because of the adverse effects on the extrahepatic organs. Consequently, with the introduction of improved venovenous bypass techniques, in situ cold hepatic perfusion has been used in selected patients since 1971. In situ hypothermic hemihepatic perfusion, introduced in 1995, prevents an ischemic insult to the contralateral hepatic lobe. Topical cooling using ice slush under total or hemihepatic inflow occlusion was reported in 1993. This technique does not require cumbersome hypothermic perfusion equipment. In attempts to minimize intraoperative bleeding by vascular occlusion, the liver surgeon must consider the benefits and technical demands of hepatic hypothermia. PMID- 9747176 TI - The history of surgery. PMID- 9747177 TI - The benefits of antibiotic prophylaxis for groin repair. PMID- 9747178 TI - Antibiotic prophylaxis and open groin repair. PMID- 9747179 TI - A combined Hansch/Free-Wilson approach as predictive tool in QSAR studies on propafenone-type modulators of multidrug resistance. AB - A series of 48 propafenone-type modulators of multidrug resistance was synthesized and their P-glycoprotein inhibitory activity was measured using the daunomycin efflux assay. Both a Free-Wilson and a combined Hansch/Free-Wilson analysis were performed using log P, partial log P and molar refraction values as Hansch descriptors. The results of the Free-Wilson analysis show that modifications on the central aromatic ring generally influence pharmacological activity, whereby in almost all cases a decrease in MDR-modulating potency is observed (Q2cv = 0.66). The combined approach results in equations with remarkably higher predictive power (Q2cv = 0.83), specifically molar refractivity shows high significance in all equations derived. This indicates that polar interactions also contribute to protein binding. PMID- 9747180 TI - Synthesis, structure, and biological activities of some N-(4,5-dihydro-1H imidazol-2-yl)-1,3-dihydrobenzimidazole derivatives. AB - A series of novel N-(4,5-dihydroimidazol-2-yl)-1,3-dihydrobenzimidazole derivatives 2a-d, 3a-d and 4a-p were prepared and their structure was determined by IR and NMR spectroscopic data as well as X-ray analysis of carbonitrile 2a. The compounds were studied as potential inhibitors of the human blood platelet aggregation induced by adrenaline or ADP. Compounds of type 3 proved efficacious for the reduction of arterial blood pressure upon intravenous administration to normotensive rats. PMID- 9747181 TI - Synthesis and immunotropic activity of some 2-aminobenzimidazoles, Part 4. AB - 2-Aminobenzimidazole reacted with selected esters of alpha, beta-unsaturated acids and alpha, beta-unsaturated ketones to form derivatives of 1,2,3,4 tetrahydropyrimido[1,2-a]benzimidazol-2-ones, 1,4-dihydropyrimido[1,2 a]benzimidazoles, and 2-acetylaminobenzimidazole. 2-Cinnamoylaminobenzimidazole, 4-phenyl-1,2,3,4-tetrahydropyrimido[1,2,-a]benzimidazol-2-on e, 4-(benzimidazol-2 ylamino)-4-phenylbutan-2-one, and 4-methyl-2-phenyl-1,2,3,4 tetrahydropyrimido[1,2-a]benzimidazole were tested for their potential activity in immunological assays in the mouse model. Compound 1, at a dose of 100 micrograms/mouse, significantly inhibited the humoral immune response and, at the same time, stimulated the cellular type of that response. The proliferative response of mouse splenocytes to concanavalin A was inhibited only at a higher dose (2 micrograms/ml). The immunotropic activity of 4, on the other hand, was uniformly strongly inhibitory in all applied tests. The suppressive activity of 4 was lower in the cellular immune response and proliferation tests than that of cyclosporin A. PMID- 9747182 TI - Synthesis and preliminary CNS depressant activity evaluation of new 3-[(3 substituted-5-methyl-4-thiazolidinon-2-ylidene)hydrazono]-1H-2- indolinones and 3 [(2-thioxo-3-substituted-4,5-imidazolidinedion-1-yl) imino]-1H-2-indolinones. AB - A series of (+/-) 3-[(3-substituted-5-methyl-4-thiazolidinon-2- ylidene)hydrazono]-1H-2-indolinones (2a-h) and 3-[(2-thioxo-3-substituted-4,5 imidazolidinedion-1-yl)imino] -1H-2-indolinones (3a-g) were synthesized by the cyclization of 3-(4-substituted-thiosemicarbazono)-1H-2-indolinones (1a-h) with ethyl 2-bromopropionate in anydrous ethanolic medium and oxalyl chloride in anhydrous diethyl ether, respectively. The structures of 2 and 3 were confirmed by analytical and spectral data (IR, 1H NMR, 13C NMR, and EIMS). The configuration of 3 was assigned on the basis of 1H NMR and 13C NMR data. 2c, 2d, 2g, 2h, and 3a-g were evaluated for anticonvulsant activity against maximal electroshock (MES) and subcutaneous pentylenetetrazol (ScMet) induced seizures. Among the compounds tested, only 2d exhibited some activity in anticonvulsant identification (Phase I) trials in mice. 2a, 2b, 2d, 2g, 2h, and 3a-g were additionally tested for potentiating effects on pentobarbital induced hypnosis in mice. All of the test compounds increased the sleeping time of pentobarbital significantly (p < 0.05) and the most potent compound was found to be 3a. PMID- 9747183 TI - Biotransformation and toxicity of the lipoxygenase inhibitor 2-hydroxy-5 methyllaurophenone oxime (FLM 5011) on Hep G2 cells. AB - 2-Hydroxy-5-methyllaurophenone oxime (FLM 5011) is an inhibitor of lipoxygenase with antiinflammatory and antiallergic actions. We incubated FLM 5011 on the human immortal cell line Hep G2 and found a similar metabolite spectrum as in rat urine and faeces. We also measured the cytotoxicity of FLM 5011 on Hep G2 monolayers by the amido black assay and found that the influence of cell proliferation is partly due to apoptotic activities. Although the metabolic activity of Hep G2 cells is lower compared to rat hepatocyte cultures they are a suitable test system for biotransformation studies. Their higher proliferation rate allows toxicity to be characterised more exactly. PMID- 9747184 TI - The neuropsychology of 3-D space. AB - The neuropsychological literature on 3-D spatial interactions is integrated using a model of 4 major behavioral realms: (a) peripersonal (visuomotor operations in near-body space), (b) focal extrapersonal (visual search and object recognition), (c) action extrapersonal (orienting in topographically defined space), and (d) ambient extrapersonal (orienting in earth-fixed space). Each is associated with a distinct cortical network: dorsolateral peripersonal, predominantly ventrolateral focal-extrapersonal, predominantly ventromedial action-extrapersonal, and predominantly dorsomedial ambient-extrapersonal systems. Interactions in 3-D space are also regulated neurochemically with dopaminergic and cholinergic excitation associated with extrapersonal activation and noradrenergic and serotonergic excitation associated with peripersonal activation. This model can help explain the 3-D imbalances in prominant neuropsychological disorders. PMID- 9747185 TI - Personality, mood, and cognitive processing of emotional information: three conceptual frameworks. AB - This article reviews evidence for the roles that mood states and personality traits play in the processing of emotion-congruent information across different cognitive tasks. Evidence is reviewed for 3 emotion-congruency frameworks, each summarizing a different route to emotional processing: the traditional approach, a moderation approach, and a mediation approach. Most of the traditional literature includes studies that examine the effects of moods and traits on emotional processing separately; these studies have yielded some inconsistent findings. The moderation and mediation approaches offer potential solutions to the lack of consistency obtained in the traditional literature by allowing for the combined effects of personality traits and mood states on the processing of emotional information. The moderation approach suggests that mood states interact with individual differences in emotion-relevant personality traits to influence emotion-congruent processing. The mediation approach suggests that personality traits predispose individuals to certain mood states, which then influence emotional processing. These approaches provide a framework for understanding the literature and a starting point for future research on emotion-congruent processing. PMID- 9747186 TI - The happy personality: a meta-analysis of 137 personality traits and subjective well-being. AB - This meta-analysis used 9 literature search strategies to examine 137 distinct personality constructs as correlates of subjective well-being (SWB). Personality was found to be equally predictive of life satisfaction, happiness, and positive affect, but significantly less predictive of negative affect. The traits most closely associated with SWB were repressive-defensiveness, trust, emotional stability, locus of control-chance, desire for control, hardiness, positive affectivity, private collective self-esteem, and tension. When personality traits were grouped according to the Big Five factors, Neuroticism was the strongest predictor of life satisfaction, happiness, and negative affect. Positive affect was predicted equally well by Extraversion and Agreeableness. The relative importance of personality for predicting SWB, how personality might influence SWB, and limitations of the present review are discussed. PMID- 9747187 TI - Learning and homeostasis: drug addiction and the McCollough effect. AB - The contribution of conditioned responses (CRs) to homeostasis may be seen by examining seemingly disparate phenomena of color vision (aftereffects and chromatic adaptation) and drug addiction (withdrawal symptoms and tolerance). Color aftereffects may be elicited by nonchromatic stimuli previously paired with color (the McCollough effect, [ME]). Similarly, pharmacological aftereffects may be elicited by nonpharmacological stimuli previously paired with a drug (withdrawal symptoms). The authors summarize evidence indicating that both the ME and withdrawal symptoms are CRs. The chromatic CR is expressed as chromatic adaptation in the presence of color, and the ME in the absence of color. The pharmacological CR is expressed as pharmacological adaptation (tolerance) in the presence of the drug, and withdrawal symptoms in the absence of the drug. Both drug withdrawal symptoms and the ME are manifestations of the contribution of conditioning to normal homeostatic regulation. The authors discuss the implications of this conclusion for understanding regulatory processes and the evolution of behavioral mechanisms. PMID- 9747189 TI - Infrahyoid neck. AB - Imaging is an indispensable tool in patients with clinical suspicion of infrahyoid neck disease. CT and MR imaging can establish a positive diagnosis by showing a true mass (versus a pseudomass). In addition, by defining the exact space of origin of the lesion and its characteristics (CT density, MR signal, homo- or heterogeneity, contour, contrast enhancement), imaging can predict the correct diagnosis. Because it offers multiplanar, multiparameter information, MR imaging, performed with a dedicated coil and appropriate artifact-reduction techniques, usually is the modality of choice. PMID- 9747188 TI - A simplified approach to the spaces of the suprahyoid neck. AB - CT and MR imaging permit exquisite visualization of the complex spatial anatomy of the suprahyoid neck. In spite of the large body of literature devoted to these spaces, their names and locations often are difficult to learn. By knowing the spaces and their components, the radiologist can generate an anatomically based differential diagnosis. This article presents a simplified approach to the various spaces of the suprahyoid neck and their anatomic components. Each space is discussed separately and is accompanied by a table that lists a differential diagnosis based primarily on the normal anatomic contents of the space. PMID- 9747191 TI - The temporal bone. Contemporary diagnostic dilemmas. AB - The entire topic of temporal bone imaging cannot be addressed in a single article. This article discusses the clinical areas in which there have been particularly important advances: inflammatory disease, sensorineural hearing deficit, pulsatile tinnitus, facial nerve dysfunction, and the postoperative temporal bone. The common thread linking those sections is an attempt to emphasize their pitfalls. PMID- 9747190 TI - Imaging of the skull base. AB - Skull-base imaging has been a key factor in the advancement of skull-base surgery. The analysis of MR imaging or CT of the skull base emphasizes important landmarks, which are key to surgical planning. Although the definitive diagnosis usually is done by biopsy, the radiologist can limit the list of possibilities of the identity of a skull base lesion. The apparent site of origin is a key factor. Separation of cystic abnormalities from more solid enhancing abnormalities also is critical. PMID- 9747192 TI - An approach to the diagnostic imaging of jaw lesions, dental implants, and the temporomandibular joint. AB - This article is an overview of three complex and often confusing areas of diagnostic imaging. It provides the reader with a starting point and a frame of reference from which to proceed when confronted with either an unknown jaw lesion, evaluating a potential dental implant patient, or a patient with TMJ internal derangement. PMID- 9747193 TI - Larynx and hypopharynx. AB - Cross-sectional imaging with CT and MR imaging plays an indispensable complementary role to endoscopy in the pretherapeutic work-up of laryngeal and hypopharyngeal cancer and is very valuable for the detection of tumor recurrence after surgery or radiation treatment as well. Close interdisciplinary co operation and the radiologist's familiarity with therapeutic options and challenges are required in order to use these powerful diagnostic tools in an optimal fashion. Other indications for cross-sectional imaging of the larynx and hypopharynx include uncommon neoplasms, benign masses, inflammatory disorders, and trauma. PMID- 9747194 TI - Sinonasal imaging. Anatomy and pathology. AB - This article provides a clear understanding of the pathophysiology of sinonasal inflammatory diseases and the rationale behind endoscopic surgery. Normal anatomy and pertinent anatomic variants that should be included in the radiology report are described. The relative role of CT and MR imaging in evaluation of inflammatory and neoplastic lesions is emphasized. PMID- 9747195 TI - Salivary glands. AB - This article discusses the anatomy, physiology, and pathology of the parotid, submandibular, and sublingual glands, which often are referred to as the major salivary glands. Overall, diseases of the salivary glands are relatively uncommon; however, as an organ system, they have the greatest diversity of pathology. Acute viral and bacterial inflammatory diseases are the most common salivary gland abnormalities; tumors are uncommon. The imaging approach to these lesions is discussed. PMID- 9747196 TI - Oral cavity and pharynx. AB - Imaging of the oral cavity and pharynx often is required in three settings: assessment of an inflammatory mass in association with odontogenic, tonsillar, or pharyngeal infections; determination of the cause of a submucosal mass; and staging of squamous-cell carcinomas. Spread of infection from the oral cavity and pharynx can lead to abscesses in the masticatory space, the retropharyngeal compartment, and in a parapharyngeal location. Submucosal masses include congenital cysts (thyroglossal and dermoid), benign neoplasms (hemangioma, schwannomas, pleomorphic adenomas juvenile angiofibromas), inflammatory cysts (mucous retention cysts, ranulas), and pseudotumors (osteophytes, carotid arteries). Staging of squamous-cell carcinomas must focus on deep invasion, spread to the brain, nerves, mandible, prevertebral muscle, and pre-epiglottic fat. PMID- 9747197 TI - Head and neck lesions. Radiologic-pathologic correlations. AB - Correlating findings on imaging studies with those on histopathologic examination can define the limitations and strengths of the radiologist's imaging armamentarium. Although CT is particularly strong in identifying the character of the matrix of a head and neck lesion, MR imaging has proved superior in the mapping of most malignant neoplasms. Sometimes the combination of CT and MR imaging characteristics of a lesion may yield a specific diagnosis; however, in most instances, the radiologic appearance is sufficiently nonspecific that aspiration cytology or biopsy is required. This article reviews the effectiveness of imaging for characterizing and outlining lesions. PMID- 9747198 TI - [Pruritus and the hepatitis C virus. The MULTIVIRC Unit]. AB - OBJECTIVE: Determine the characteristics of pruritus in a selected population of patients with positive hepatitis C virus serology. PATIENTS AND METHODS: In a retrospective study, we re-examined outpatient reports for patients who consulted for pruritus from January 1993 to April 1996 and were followed in a hepatology unit for hepatitis C infection. The following data were collected: age, sex, risk factors, HIV and HBV serologies, duration of pruritus and diagnosis, ALAT, gamma GT and total bilirubin, METAVIR score, HCV RNA PCR, search for cryoglobulins, antiviral and dermatology treatments. There were 1,060 patients followed in the hepatology unit during this period, including 327 with cirrhosis. RESULTS: Twenty seven patients were retained for study, 16 men and 11 women, mean age 53 years. None of the patients had HIV infection, 7 had a past history of hepatitis B infection (positive for anti-Hbc antibodies). Median duration of pruritus prior to consultation was 3 months (95 p. 100 CI: 3 months-2 years). Pruritus was associated with non-specific lesion in 19 cases (70 p. 100: 95 p. 100 CI: 51-85 p. 100). There were excoriated eczema-like lesions in 11 cases (41 p. 100: 95 p. 100 CI: 15-49 p. 100). Other causes were urticaria in 5 cases (18 p. 100: 95 p. 100 CI: 7-36 p. 100), including 1 case of urticarial vasculitis with cryoglobulinemia, 2 cases with atopic dermatitis, and 1 case of primary biliary cirrhosis. In four cases, lichen planus was associated. Skin biopsies were obtained in 10 patients and showed eczema-like alterations in 9 and urticarial vasculitis in 1. Mean ALAT and gamma GT levels were 2.6 N (95 p. 100 CI: 1.9 N 3.3 N) and 2.2 N (95 p. 100 CI: 1.4 N-3 N) respectively, including 11 cases without cholestasis (normal gamma Gt). PCR was positive in 13 cases out of 15. Cryoglobulinemia was found in 10 cases out of 24. At consultation, 3 patients were given ursolvan, 7 interferon, 1 ursolvan with ribavirin, and 3 an interferon ribavirin combination. Dermatology treatment associated antihistamine agents, emollients, and corticosteroids. This population of hepatology patients referred for pruritus comprised 2.5 p. 100 of all patients (95 p. 100 CI: 1.7-3.6 p. 100). Among them, 1.8 p. 100 (95 p. 100 CI: 1.1-2.7 p. 100) had eczema-like lesions associated with cutaneous xerosis. DISCUSSION: Pruritus in our patient population was generally associated with non-specific excoriations, prurigo or xerosis in 19 cases (70 p. 100). As only ambulatory patients were retained for analysis, this is not a comprehensive population and the percentage of non-specific pruritus, evaluated at 1.8 p. 100, is probably an underestimation. Cholestasis cannot explain alone these manifestations since 11 patients had normal gamma GT levels. Several etiologies could be involved: a direct effect of HCV, interferon. A prospective study should allow an estimation of frequency, risk factors and possible impact on quality of life. PMID- 9747199 TI - [Erythroderma with immunoglobulin deposits along the basal membrane. Pemphigoid erythroderma?]. AB - INTRODUCTION: There are many clinical forms of bullous pemphigoid. Nodular, localised, vegetating, vesicular and dysidrosiform variants have been described. The appartenance to the spectrum of pemphigoid was confirmed by the demonstration of antibodies directed against the 230 and 180 kDa BP antigens. Three cases of erythrodermic pemphigoid were recently described. OBSERVATIONS: We describe two cases of patients suffering from erythroderma for several months associated with a linear deposit of immunoglobulins and complement along the basement membrane. In the first observation, the patient suffered from bullae of the legs before developing erythroderma. The positive direct immunofluorescence of the skin was associated with circulating anti basement membrane antibodies, making therefore the diagnosis of erythrodermic pemphigoid possible. In the second case, the patient never developed blisters. We also found a linear deposit along the basement membrane, localized on the dermic side of a skin treated by molar NaCl. No circulating antibodies were found by indirect immunofluorescence or by immunoblott. Direct immuno electron microscopy showed a deposit of immunoglobulins into the deep layers of the lamina lucida and into a part of the lamina densa. CONCLUSION: These two cases suggest that there are true erythrodermic pemphigoid without blisters. The value of the systematic direct immunofluorescence examination of the skin of erythroderma should be evaluated. PMID- 9747200 TI - [Actinic prurigo of childhood. 3 familial cases associated with HLA-DR 0407]. AB - BACKGROUND: Actinic prurigo, as idiopathic skin reaction involving light-exposed areas, was first described in American Indians. Actinic prurigo was early considered to be a particular form at polymorphous phototoxicity, but can be identified as a specific entity on the bases of clinical features and epidemiological characteristics. CASE REPORTS: Three children in the same family developed photosensitive reactions early in childhood with characteristic polymorphous and persistent eczema-like or papulo-nodular pruriginous lesions which predominated in light-exposed areas and appeared several hours after exposure to sun. The lesions persisted during the winter season. The lesions followed a chronic course but tended to improve at puberty. Clinical laboratory tests, serum and urine porphyrin levels and antinuclear factors were normal. Histology and photobiology explorations gave non-specific results. DISCUSSION: These observations have three points in common with actinic prurigo observed in American Indians. HLA typing showed that our three patients, as in white patients in Great Britain, had a significant association with a specific HLA DR1 subtype: DRB1*0407. This DRB1*0407 alleles could play a role in initiating the immune response to a light-induced peptide antigen. This particular genetic predisposition, if confirmed in other studies, would be an additional argument for distinguishing actinic prurigo as a specific polymorphous phototoxicity entity. PMID- 9747201 TI - [Lichenoid contact dermatitis from the ink of a red pen]. AB - INTRODUCTION: Numerous cutaneous inflammatory reactions have been reported in literature, after using red dyes; most of them are lichenoid reactions and occur after tattooing. Few reports have mentioned inflammatory complications after using red ink. CASE REPORT: A 21-year-old woman was examined for papular erythematous, pruritic lesions on the back of her left hand. They developed within the area of a red inked pen writing, a few days before. There was an isomorphic reaction on the cheek which was in contact with the hand during sleep. Diagnosis of lichen reaction to red ink was made, and histologically confirmed on the hand. DISCUSSION: We report the first case to our knowledge of lichen reaction to red inked pen, which can be compared to hypersensitivity reactions to red pigment reported in literature. A Koebner phenomenon by friction, usual with lichen, is possible, but doesn't explain the reaction on the cheek. A contact dermatitis to red pigment of ink can also be suggested, although mostly eczema like reactions have so far been reported; it seems moreover that lichenoid reactions reported only occur after intradermal injection of pigment during tattooing. This contact mechanism would nevertheless explain both localizations of lichen reaction in our patient. PMID- 9747202 TI - [Werner's syndrome]. AB - BACKGROUND: Werner's syndrome associates early aging in young adults, small height, cataract, glucose intolerance, hypogonadism, skin ulcers, vascular calcifications and osteoporosis. CASE REPORT: We report a new case of Werner's syndrome in a 34-year-old man with suggestive alterations of the skin and endocrine anomalies in addition to hypospadias, urethral stenosis, bilateral mega ureter and chronic renal failure. DISCUSSION: The diagnosis of Werner's syndrome in our patient was unquestionable because of the clinical presentation and the familial context. However, the urology anomalies have not been reported in this syndrome. A simple coincidence cannot be excluded. PMID- 9747203 TI - [Photosensitive prurigo in AIDS]. AB - INTRODUCTION: A case of photosensitive prurigo during AIDs is reported. This is the second case in the literature. We discuss the relations between HIV infection and photodermatoses. OBSERVATIONS: A woman, known to be HIV seropositive from 1990, developed during the spring 1990 a prurigo on light exposed areas who received the next year. A photobiological investigation was performed, showing a polymorphic light eruption induced by UVB. DISCUSSION: Patients infected with HIV have a high prevalence of UV radiation responsive skin diseases. On the other hand, UVA radiations, UVB and UVC have been shown to induce activation and replication of HIV. PUVA therapy and UVB therapy have shown their efficacity in the treatment of many photodermatoses associated with HIV infection, without any worsening of the illness. Many questions are not yet solved in the relationship between HIV and photosensitivity and the photobiological investigation should be more frequently done. PMID- 9747204 TI - [Multiple isolated cutaneous myxomas]. AB - INTRODUCTION: Cutaneous myxomas are rare. They are more often single; when they are multiple, they may be one of the component of the Carney's syndrome. CASE REPORT: We report a case of multiple and isolated cutaneous myxomas arising at 19 year-old. Diagnosis was confirmed by histologic studies. Examination failed to reveal other cutaneous or visceral features. DISCUSSION: The multiplicity of cutaneous myxomas is an essential element for diagnosis of Carney's syndrome; other components and notably atrial myxomas can appeared over time. Normal visceral explorations don't eliminated the diagnosis, and follow-up including screening echocardiography must be ensured before concluding, as in our case, as multiple and isolated cutaneous myxomas. PMID- 9747205 TI - [Pretibial epidermolysis bullosa. A rare form of epidermolysis bullosa simplex]. AB - INTRODUCTION: Pretibial epidermolysis bullosa had been classified as a rare localized form of autosomal dominant dystrophic epidermolysis bullosa. OBSERVATION: We report a sporadic case of a patient suffering from bullous lesions induced by minor trauma on pretibial skin. The lesions healed with atrophic scars. No milia formation was observed. The mapping of dermoepidermal junction by LH 7:2 and GB3 monoclonal antibodies was normal. By electron microscopy, numerous perinuclear vacuoles were observed and the cleavage occurred within the basal keratinocytes. DISCUSSION: This patient had clinical features in accordance with a diagnosis of pretibial epidermolysis bullosa. However, in contrast to previous case reports, the ultrastructural pattern was this of an epidermolysis bullosa of simplex type. PMID- 9747206 TI - [Atypical manifestations in familial type 1 Waardenburg syndrome]. AB - BACKGROUND: Waardenburg syndrome is an uncommon genetic disorder. Four clinical types are recognized. Three responsible genes have been identified (PAX 3: for type I syndrome, MITF and EDN3 for types II and IV respectively). CASE REPORT: We report the case of a patient with Waardenburg type I morphotype who had atypical neurological manifestations. Decisive elements for diagnosis were the presence of Waardenburg syndrome in the family and, in affected kin, a mutation causing a shift in PAX 3 gene reading. DISCUSSION: This case confirms the variability of Waardenburg signs within one family. The association of unusual neurological manifestations in the proband suggested that Vogt Koyanagi Harada disease may have been associated and may show some relationship with familial Waardenburg syndrome. PMID- 9747207 TI - [Erythema multiforme caused by saquinavir]. AB - BACKGROUND: Saquinavir is a protease inhibitor used for the treatment of HIV infection. Adverse skin reactions have been rare. We report here the first case of erythema multiforme in a patient given saquinavir. CASE REPORT: A 32-year-old man was seropositive for HIV and consulted due to the development of round maculo papular lesions centered on a bulla and two erosive lesions of the palate five days after the introduction of saquinavir. Histology was compatible with erythema multiforme. After withdrawal of saquinavir, the skin and mucosal lesions regressed in 15 days, with no recurrence at 3 months. DISCUSSION: Adverse skin reactions to saquinavir are exceptional (eruptions, pruritus). We describe here the first case of erythema multiforme caused by saquinavir (imputability criteria 12 BO). Due to the structural analogy of saquinavir with other protease inhibitors (indiravir, ritonavir, nelfinavir) it would be difficult to prescribe a compound of the same class. PMID- 9747208 TI - [Acquired tufted angioma in an adult]. AB - INTRODUCTION: Acquired vascular tumors have been raising new interest since Kaposi's illness was discovered in human immunodeficiency virus infection. We herein report a case of a rare different entity of acquired vascular tumor, described by E. Wilson-Jones for the first time in 1976. CASE REPORT: A 69-year old woman had had for several months an asymptomatic erythematous lesion on the face gradually increasing in size. Histopathologic findings were pathognomonic of "tufted" angioma. The patient did not receive any treatment because of uncertain efficiency and because her lesion was well tolerated. DISCUSSION: "Acquired tufted angioma" is a benign angioma of the skin mainly occurring in children and young adults of both sexes. Histopathologic findings are pathognomonic. Typically, tufted angioma enlarges for a few years and then ceases growing and remains stable. Spontaneous regression may occur. The principal differential diagnoses are Kaposi's illness and low grade malignant angiosarcoma. Pulsed dye laser in the most efficient treatment. PMID- 9747209 TI - [Acquired tufted angioma in an adult: failure of pulsed dye laser therapy]. AB - BACKGROUND: Tufted angioma, described by Wilson Jones in 1976, is a benign acquired vascular tumor occurring in children or young adults, usually located on the neck or the upper part of the thorax. Pathology examination confirms the diagnosis showing well-limited lobules in the dermis composed of tight clusters of capillaries without atypical cells. CASE REPORT: An 81-year-old woman consulted for a large extensive angiomatous lesion involving the neck and shoulder which had developed over two years. Histopathology reported tufted angioma. Treatment with pulsed dye laser was unsuccessful. DISCUSSION: Different treatments have been proposed for tufted angiomas: surgery, cryotherapy, interferon, argon laser and pulsed dye laser. This is a second cases of unsuccessful treatment, perhaps due to deep extension of the angioma. PMID- 9747210 TI - [Maffucci syndrome associated with epidermoid carcinoma of the nasopharynx]. AB - BACKGROUND: Maffucci's syndrome is a dysembryoplasia of the mesoderm, explaining the dual involvement of cartilage and vascular tissue. The risk of malignant degeneration or associated tumors is high in this uncommon disease. We report a case of Maffucci's syndrome associated with squamous cell carcinoma of the cavum. CASE REPORT: A 37-year-old man consulted for multiple angiomas and chondromas which had developed since childhood. The diagnosis of Maffucci's syndrome was obvious. Radiological exploration of the limbs confirmed chondromatosis and biopsy of the cavum performed because of the occurrence of epistaxis, revealed squamous cell carcinoma. DISCUSSION: Maffucci's angiochondromatosis is a rare, non-hereditary but sometimes congenital disease. Angiomas predominate on the hands and feet. Sarcomatous degeneration is the main complication of these chondromas. Angiomas rarely become malignant, but when they do are generally more aggressive than chondromas. The frequency of neoplasia in Maffucci's syndrome would suggest that there is a supplementary oncogenic factor. Several types of malignancy associated with Maffucci's syndrome have been reported, but to our knowledge there has been no publication concerning an association between Maffucci's syndrome and squamous cell carcinoma of the cavum. PMID- 9747211 TI - [Cutaneous pseudolymphoma caused by carbamazepine]. AB - INTRODUCTION: Drug-induced cutaneous pseudolymphomas are poorly reported. They clinically and pathologically mimic malignant lymphomas but their evolution is benign after drug withdrawal. CASE REPORT: We herein reported a case of carbamazepine++-induced pseudolymphoma in a 30 year-old woman of particular interest because of the paucity of clinical symptoms (only six papules of less than one centimetre each). COMMENTS: Phenytoins, carbamazepine++, barbiturates and ACE inhibitors are the main drugs inducing cutaneous pseudolymphomas. These usually mimic T-cell lymphomas. The clinical spectrum of cutaneous pseudolymphomas is broad including severe aspects, such as erythroderma or tumoral eruption and benign appearing one as in our case. Drug investigation is mandatory for each patient with lymphoma appearing cutaneous infiltrates because dramatic and definitive healing of the lesion is always expected in case of pseudolymphoma. PMID- 9747212 TI - [Chronic depigmentation due to positive patch tests for methacryate derivatives]. AB - INTRODUCTION: Many chemical products are known to induce depigmentation. This phenomenon was never reported with methacrylates which are components of acrylic resine. CASE REPORT: A female patient with artificial nails developed contact dermatitis. Localized depigmentation at the site of positive patch tests to methacrylates derivatives was observed. DISCUSSION: The chemical substance could have a direct influence either by its toxic effect or by the induced inflammatory reaction. PMID- 9747213 TI - [Case for diagnosis. Neuro-sarcoidosis]. PMID- 9747214 TI - [Case for diagnosis. Keratosis follicularis]. PMID- 9747215 TI - [Epidermotropic cutaneous T cell lymphomas]. PMID- 9747216 TI - [Epidemiologic concepts useful to dermatologists]. PMID- 9747217 TI - [Use of ultraviolet rays in childhood]. PMID- 9747219 TI - [Rare autochthonous parasitic diseases in dermatology]. PMID- 9747218 TI - [Prevention of facial herpetic infections after chemical peel and dermabrasion: new treatment strategies in the prophylaxis of patients undergoing procedures of the perioral area]. PMID- 9747220 TI - [Epstein-Barr virus research by in situ hybridization in 65 cutaneous T cell epidermotropic lymphomas]. AB - BACKGROUND: The Epstein-Barr virus (EBV) is a highly mutagenic virus known to be the cause of several types of lymphoma. There has been some controversy concerning EBV in cutaneous T-cell lymphomas. The aim of this study was to search for EBV with a sensitive method: in situ hybridization in 65 patients with cutaneous T-cell lymphomas. PATIENTS AND METHODS: From 1990 to 1995, 158 samples from 65 patients with cutaneous T-cell lymphoma (2 stage IA, 12 IB, 4 IIA, 29 IIB, 16 Sezary syndrome, 2 stage IV) were collected. In situ hybridization with EBER and Bam W probes recognizing the viral latency genes were used to search for EBV. RESULTS: EBV was evidenced with at least one of the two probes in 43 samples (26 p. 100). Prior to alpha interferon treatment, 18 p. 100 of the samples were positive for EBER compared with 18 p. 100 for Bam W. After alpha interferon treatment, there was a significantly higher percentage of EBER positive samples (39 p. 100; p = 0.03). Inversely, there was no difference for the Bam W probe (p = 0.2). Clinical stage had no effect on the presence of EBV (p = 0.18). CONCLUSION: Our series evidenced the variable presence of EBV, identified by in situ hybridization, in cutaneous T-cell lymphoma. Few infiltrating cells are infected. This would be an argument in favor of an indirect role of the EBV in the transformation process. In addition, alpha interferon increases the life time of EBERs, sensitizing detection of this latency gene. PMID- 9747221 TI - [Muscular damage during isotretinoin treatment]. AB - BACKGROUND: The effect of isotretinoin on muscle is considered to be uncommon and benign. We analyzed the files of all our patients given isotretinoin over a 5 year period and determined the incidence and gravity of its effect on muscles. MATERIALS AND METHODS: Sixty treatments with isotretinoin were studied. Myalgia and elevated CPK observed at the pretherapeutic consultation and each month or 2 months thereafter were recorded. RESULTS: Muscle symptoms were noted in 60 p. 100 of the cases: myalgia in 51 p. 100 and elevated CPK in 41 p. 100. In this group, male sex (H/F = 2.6) and sports activities (70 p. 100) were found more often. In 5 patients, CPK level was 5 times the normal, defining rhabdomyolysis. One patient interrupted treatment because of persistently high CPK levels. DISCUSSION: Myalgia and elevated CPK signal skeletal muscle involvement. These signs were more frequent in our series than in reports in the literature, probably because we systematically looked for them. Besides use of isotretinoin, one case of viral infection and sports activities, no other explanatory cause could be found. Isotretinoin could have a potentializing effect on other myotoxicity inducers (drugs, infection, fever, muscular exertion...). The benign nature of the muscle effect appears to be validated although there were some patients who had persistent and/or severe signs. PMID- 9747222 TI - [Absence of HHV-8 virus detected in immature hemangiomas in infants]. AB - INTRODUCTION: The aim of our study was to search for the presence of HHV-8 DNA sequences in Biopsy specimens from hemangioma of the infancy. MATERIAL AND METHODS: The study included 9 biopsies from hemangioma. DNA of human beta-globin gene and HHV-8 were searched for by PCR using specific primers. Amplified products were revealed after an hybridization with an internal probe digoxigenin labelled. RESULTS: Human beta-globin gene could be detected in all samples illustrating the absence of PCR inhibitor. HHV-8 could never be detected in samples analyzed. DISCUSSION: Our study does not imply any causative role of HHV 8 in the pathogenesis of hemangioma. This result must be confirmed by serologic studies. PMID- 9747223 TI - [Serum and salivary immunoglobins A in atopic dermatitis. Prospective and comparative case control study]. AB - IgA system has been poorly studied in patients with atopic dermatitis (AD). Previous studies have showed that a transient serum IgA deficiency in infancy could lead to atopic disease. In addition, decrease in salivary IgA has been demonstrated in patients with AD. The purpose of our work was to study the IgA system both in serum saliva in patient with AD. PATIENTS AND METHOD: We conducted a controlled prospective study from January 1994 to May 1996. 46 patients with AD and 52 healthy volunteers matched for sex and age were included. Atopic patients fulfilled at least three major and three minor features defined by Hanifin and Rajka. None above atopic criteria were present in the control group. Saliva was collected using a small cylinder of a cotton-wool-like substance (Salivette) kept in the buccal fold. Serum and saliva samples were assayed for IgA using standard nephelometric method and time-resolved immunofluorometric assay. Secretory IgA were assayed by a sandwich-type enzyme linked immunosorbent assay. Blood eosinophils and serum IgE were also evaluated. RESULTS: IgA and secretory IgA were detected in all serum and saliva collected. No statistically significant difference were observed in serum or in saliva for both IgA and secretory IgA between patients with AD and controls. As expected, blood eosinophils and serum IgE were significantly increased in patients with AD. DISCUSSION: None patients (atopic or control) exhibited IgA deficiency. Although no statistically significant, a trend to higher concentrations of serum and salivary IgA was observed in patients with AD suggesting a stimulation of mucosa-associated lymphoid tissue in these patients. PMID- 9747224 TI - [Cutaneous dirofilariasis from Dirofilaria repens. A case contracted in Gironde]. AB - INTRODUCTION: Cutaneous dirofilariasis caused by Dirofilaria repens is a parasitic disease in dogs and cats. It is exceedingly rare in man who is an occasional host. The parasite fails to reach maturity. The true incidence of dirofilariasis is probably greater than recognized. This can be explained by the apparent benign nature of the lesions that may not warrant excision. CASE REPORT: A 12-year-old girl presented a cutaneous nodular lesion involving the forehead. Surgical excision allowed diagnosis: cutaneous dirofilariasis caused by Dirofilaria repens. The child had always stayed in mainland France. Histopathological examination showed the worm trial with inflammatory granulomatous reaction of the deep dermis. The parasite extraction allowed successful treatment. DISCUSSION: This new case of cutaneous dirofilariasis emphasizes that this parasitic disease is present in mainland France. The infection is more common in Southern France. The present report describes a pediatric case in Gironde, where it is exceedingly rare. Microscopic examination identified, and it was unusual, a male worm. PMID- 9747225 TI - [Salmonella enteritidis septicemia manifesting as a suppurated thrombophlebitis]. AB - BACKGROUND: Non-typloid salmonella can cause septicemia and extradigestive disorders in immunodepressed adults. These frequent diseases can be life threatening. CASE REPORT: A 76-year-old woman was treated with corticosteroid therapy for 1 year for suppurated thrombophlebitis of the right greater saphenous vein. Weight loss, fever at 41 degrees C and Salmonella enterididis isolated from blood cultures and skin samples led to the diagnosis of septicemia with multiple septic foyers including the venous endothelium and surrounding soft tissue. DISCUSSION: In Western countries, there has been an uprise in the frequency of low-grade salmonella infections by food poisoning usually causing acute diarrhea. S. enterididis can also cause severe infectious syndromes with multiple septic localizations, main in patients with a compromised immune reaction. In our cases, Salmonella enteritidis septecemia was revealed by an unusual situation. In the literature, inaugural signs usually involve the heart or arteries, but our patient had isolated foyers involving the superficial venous network. This is exceptional especially since there was no iatrogenic venous catheter insult. For our patient, favoring factors were the long-term corticosteroid therapy and altered venous network. The portal of entry could not be clearly identified but the discovery of a sigmoid diverticulosis would be an argument favoring a digestive origin. Medical and surgical management with resection of the necrosed tissues and two adapted antibiotics in a long-term regimen led to a successful outcome. PMID- 9747226 TI - [Cutaneous manifestations during disseminated trichosporonosis in an AIDS patient]. AB - BACKGROUND: Trichosporon beigelii, causal agent of white piedra can cause disseminated infection in immunodepressed subjects. Systemic infections due to this pathogen have been reported mainly in neutropenic patients and rarely in AIDS patients. CASE REPORT: A 36-year-old HIV+ man from Senegal was hospitalized for fever and meningoencephalitis associated with skin lesions. T. beigelii was isolated from skin biopsies and cerebrospinal fluid cultures. The patients was treated with amphotericin B with regression of the skin lesions. The diagnosis of disseminated T. beigelii infection was retained. DISCUSSION: Disseminated T. beigelii infections are known to occur in immunodepressed subjects, especially in case of neutropenia. In our patient, the presence of two proven localizations (meninges and skin) and the favorable outcome with amphotericin B favored disseminated infection. The good response to treatment can probably be explained by the absence of neutropenia. Skin lesions are frequent, usually occurring as disseminated papulae or purpural nodules. Pathology examination and skin biopsy culture can provide rapid diagnosis allowing appropriate treatment. PMID- 9747228 TI - [Multiple congenital smooth-muscle hamartomas]. AB - BACKGROUND: Smooth muscle hamartoma is an uncommon lesion. Diagnosis is usually made at birth in infants presenting a plaque with minimal or no infiltration and covered with long dark hairs. Congenital forms with multiple plaques are rarely reported. CASE REPORT: A 5-day-old infant (normal pregnancy and delivery) had plaques localized on the buttocks, the left thigh, leg and shoulder and the right ankle. The plaques were minimally infiltrative and covered with long black hairs. Histology examination showed hyperplastic smooth muscle bundles with varying orientation. The diagnosis was smooth muscle hamartoma. The rest of the clinical examination was normal. CONCLUSION: This case of congenital smooth muscle hamartoma showed a particular form with partially regressive multiple plaques. PMID- 9747227 TI - [Allogeneic bone marrow transplantation in congenital erythropoietic porphyria. Gunther's disease]. AB - INTRODUCTION: The congenital erythropoietic porphyria (Gunther's disease) (CEP) is a rare autosomal recessively metabolic disease due to the deficit of uroporphyrinogen III cosynthetase, fourth enzyme of the porphyrin-heme biosynthesis. This disease is characterized by severe cutaneous photosensitivity with profound skin lesions, hemolytic anemia and excess of uroporphyrin I excretion. The vital prognosis is very bad and until now, no treatment seems to be efficient. Bone marrow transplantation seems to be able to correct the enzymatic deficit that causes the disease because it is located in the bone marrow. OBSERVATION: We report the case of a four and a half year old girl who received an allogeneic bone marrow transplantation (BMT) at the age of two. Despite an encouraging result, the first transplantation failed. A second allogeneic transplantation was attempted eight months later with the same HLA identical heterozygous donor and bone marrow engrafment succeeded. Twenty one months after the second bone marrow transplantation, clinical and biological results are still excellent. DISCUSSION: No classical treatment of CEP really proved its efficiency and no one was curative. CEP resulting from an homozygous deficiency in uroporphyrinogen III cosynthetase, enzyme that takes part in the porphyrin-heme biosynthesis which is principally located in the erythropoietic system of the bone marrow, substitution of this defective lineage by BMT was a very attractive treatment to correct this anomaly. The first bone marrow transplantation attempted on an affected child in 1990 in Manchester failed because the patient died of infections complications. After the failure of the first transplantation, our little patient is now healed twenty one months after the second BMT and biochemical anomalies are corrected. If a long follow up is necessary to appreciate the long-term efficiency of this treatment, allogenic bone marrow transplantation seems to cure Gunther's disease and must be proposed as the treatment of this affection. PMID- 9747230 TI - [Langerhans histiocytosis and acute monoblastic leukemia type LMA4]. AB - INTRODUCTION: Cutaneous manifestations of myelodysplastic syndromes are rare and polymorphous. They can be the direct expression of the hematological disease or represent signs of a vasculitis or a neutrophilic syndrome. Myelodysplastic syndromes progress sometimes toward an acute leukemia. OBSERVATION: A 53-year-old woman suffering from myelodysplastic syndrome presented for several months a cutaneous vasculitis without any histological specificity. In time, such presentation was complicated by the simultaneous occurrence of a cutaneous Langerhans cell histiocytosis and an acute monoblastic leukemia type LMA 4. The disease was rapidly fatal. DISCUSSION: The complication of a myelodysplastic syndrome by concurrent Langerhans cell histiocytosis and acute monoblastic leukemia suggests a pathogenic relationship between these two latter disorders. PMID- 9747229 TI - [Bullous lichen sclerosus after radiotherapy]. AB - BACKGROUND: Bullous lichen sclerosus is an uncommon observation after radiotherapy and can be misdiagnosed as a radiodermitis or recurrence of the neoplasia. CASE REPORT: Two women developed bullous lichen sclerosus after radiotherapy. The delay after radiotherapy was 4 months and 10 years respectively. Irradiation dose was 60 and 64 grays. The lesions covered more than the irradiated zone in one case. Stabilization or regression of the lesions was obtained with cases I topical corticosteroids, with acitretine in one case. DISCUSSION: These cases, and similar cases reported in the literature, underline the fact that the condition has only been reported in women. This is probably because of the nature of the neoplasias treated (breast cancer in 20 cases and cervical cancer in 1). The delay to onset of the skin lesions are quite variable. Lichen sclerosus or morphea may be observed in the irradiated zone but may also appear at a distance. These lesions are not associated with recurrence of the initial cancer. As no association between breast cancer and localized sclerodermia has been found, the causal role is probably played by radiotherapy, producing a Koebner phenomena in predisposed tissue. PMID- 9747231 TI - [Acyclovir-resistance zona in a immunocompromised HIV seronegative patient]. AB - INTRODUCTION: Resistance to antiviral therapy is getting actually more frequent. Immunocompromised host are more concerned with this problem. OBSERVATION: We present a case of disseminated zoster resisting to acyclovir (ACV) therapy, but healing with foscarnet in a man treated with chemotherapy for lymphoma and seronegative for HIV. CI50 of VZV strain was 48 microM for ACV, which was 2.8 times higher than value of the reference OKA strain tested simultaneously, which confirmed the resistance for ACV. DISCUSSION: Immunocompromised patients often present varicella zoster virus (VZV) infection. They usually heal in response to ACV therapy, but some HIV infected patients have already presented with resistant strains of VZV. This case is the first described in a non-HIV infected patient. Foscarnet therapy resulted twice in complete healing because of its direct activity on viral DNA polymerase, so it is efficaceous therapy for patients with thymidine-kinase-deficient ACV-resistant VZV infection. PMID- 9747232 TI - [Blue nevus of the scalp associated with a meningeal melanocytoma]. AB - INTRODUCTION: Headache opposite to a blue nevus of the scalp can reveal intra cranial melanotic lesions. CASE REPORT: A 25-year-old man caucasian was admitted to hospital for a first generalized tonic-clonic seizure. For six months, he has had episodic frontal-temporal right headache opposite to a blue pigmentary cutaneous congenital lesion in frontal territory (histology confirmed benign blue nevus). Neurologic examination noted a right congenital hereditary ophtalmoplegia. Cerebral MRI showed a right rolandic tumor with diffuse leptomeninge infiltration. This patient was operated of a meningeal melanocytoma with leptomeninges melanosis. DISCUSSION: The apparition of headache related to a blue nevus must lead to realize a cerebral MRI to look for a neuroectodermic hamartoma: melanotic tumor (in particular melanoma), or leptomeninges melanosis with high potential of degeneration. Meningeal melanocytoma is a rare benign spinal or intra cranial melanotic tumor. PMID- 9747233 TI - [Infantile acne and isotretinoin]. AB - We report a case of severe infantile cystic acne, which required a treatment with oral isotretinoin. CASE REPORT: The patient, a 22-month-old boy, presented cystic acne since 6 months of age. The child was otherwise healthy and endocrine evaluation was normal. Previous treatments had no effect, oral isotretinoine led to cure without secondary effects. COMMENTS: The etiology of infantile acne is not clearly defined, but may result from a persisting androgen-driven stimulation of sebaceous glands. Oral isotretinoin is safe and effective in case of recalcitrant infantile acne, but close monitoring is necessary because of the well-known side effects of oral retinoids. PMID- 9747234 TI - [Case for diagnosis. Disseminated miliary lupus of the face]. PMID- 9747235 TI - [Case for diagnosis. Multiple symmetrical Launois-Bensaude lipomatosis]. PMID- 9747236 TI - [Physiology and physiopathology of the epidermal unit of melanization]. PMID- 9747237 TI - [History and nosology of rheumatoid purpura]. PMID- 9747238 TI - [Darier-Ferrand tumor. Cytogenetic status]. PMID- 9747239 TI - [What is a hamartoma?]. PMID- 9747240 TI - [Plasma tryptophan bioavailability during chronic urticaria]. AB - OBJECTIVES: Chronic idiopathic urticaria is known to have psychogenic component with a triggering or favoring effect. Different tests or evaluation scales have been unable to identify a specific psychological profile. Erythrocyte-specific membrane transport of tyrptophan (TRP), the main plasma precursor of cerebral serotonin synthesis, controls, by a erythrocyte-specific storage and release mechanism, circulating TRP homeostasis. Bioavailability of circulating TRP is a factor controlling serotonin synthesis in the brain. An evaluation of the rate of TRP transfer could be a biochemical approach to chronic urticaria more informative than psychological tests. PATIENTS AND METHODS: A kinetic study of L TRP influx into circulating erythrocytes was conducted in 17 patients with chronic urticaria with no detectable cause and in 35 healthy controls. Blood samples were marked with 3H-TRP. Maximum L-TRP-specific influx (Vmax) was expressed in mumol/cell/min. The urticaria patients also underwent psychological testing to determine anxiety and depression scores using standardized scales (Hamilton). RESULTS: Mean Vmax was not significantly difference between the two groups. Vmax values were quite similar in all the control subjects but showed wide dispersion in the urticaria group. Three subgroups were found in the urticaria patients depending on Vmax: those with Vmax equivalent in control levels (+2 SD), those with Vmax less then 2 SD (29% of the patients) and those with Vmax greater than 2 SD of control levels (23% of the patients). Thus more than 50% of the urticaria patients had perturbed erythrocyte-specific L-TRP influx. The anxiety and depression scores obtained from the psychological evaluation were not correlated with Vmax. DISCUSSION: Erythrocyte-specific TRP membrane transport, evaluated by Vmax. Would not appear to be perturbed in chronic urticaria. Even though the urticaria patients could be divided into three groups according to their Vmax, the mean value was not significantly different from that in controls. These findings do not allow a conclusion concerning a perturbation of bioavailability of plasmatic TRP and any possible central serotoninergic dysfunction in chronic urticaria. PMID- 9747241 TI - [Tinea capitis in Creteil. Trends over ten years]. AB - INTRODUCTION: We analyzed tinea capitis data in a Paris suburban area over a 11 year period from (1985-1995) to evaluate epidemiology trends. PATIENTS AND METHODS: The following data were collected for patients seen at the Creteil myco dermatology clinic with cultures positive for tinea capitis: sex, age, ethnic origin, fungal culture. RESULTS: Tinea capitis was observed in 336 cases (56 p. 100 females). Eight percent of the patients were under the age of 10 years and 11 p. 100 over 20 years. Trichophyton soudanense was isolated in 45 p. 100 of the patients. Anthropophilic agents rose over the 10 year period while the number of zoophilic agents remained stable. Specific dermatophytes appeared to predominate in populations of different ethnic origin. There was a two-fold increase in the number of tinea capitis cases in the 1990-1995 period compared with the five previous years. DISCUSSION: The percentage of adults with tinea capitis (11 p. 100) is higher than the 5 p. 100 reported in the literature. The rise in the number of anthropophilic tinea capitis cases resulted from an increase in T. soudanense (originating in Africa), probably related to the increasing immigrant population. This agent was identified in 95 p. 100 of the patients of African origin. Differing lifestyles and transmission between school children makes it quite difficult to interpret the correlation between ethnic origin and specific dermatophytes. PMID- 9747242 TI - [Severe hemangiomas treated with interferon alpha-2b: seven cases]. AB - INTRODUCTION: High-dose corticosteroids are the primary means of controlling alarming hemangiomas. However, only 2/3 of these life-threatening hemangiomas regress or are stabilized with corticosteroids: and for others there is no regularly safe and effective treatment. Contradictory publications have been reported about the efficacy of interferon (INF) alpha in hemangiomas, we report our experience concerning the management of 7 cases. PATIENTS AND METHODS: Seven infants have been treated with INF alpha-2b in our Pediatric Dermatology Unit. All hemangiomas were corticosteroid resistant. Hemangiomas were located on head and neck in 6 cases and on lower limb in one case in association with platelet trapping syndrome. Mean age at beginning of treatment was 7.5 months and mean duration of treatment was 5 months. INF alpha-2b was given daily in subcutaneous injections of 3 millions units per square meter of body surface area. RESULTS: After one month of treatment, 2 cases of facial hemangiomas showed a dramatic improvement, and INF alpha-2b was maintained for one year. One case was stabilized, but INF was ineffective in 3 cases after 2 months of treatment (the child with platelet trapping syndrome died at 4 months of age). The treatment had to be stopped in one case because of hepatitis. No long term side effects were noted with a mean follow-up of 27 months. CONCLUSION: INF alpha can be an interesting alternative therapy for infantile hemangiomas. However, only a minority of patients can be considered as excellent responders at current dosages and long term secondary effects cannot be excluded. PMID- 9747243 TI - [Epithelioid sarcoma manifesting as chronic plantar arch ulceration]. AB - BACKGROUND: Epitheloid sarcoma is an uncommon malignant soft tissue tumor observed in the distal extremities of young men. We report a case of long standing ulceration of the sole which was found to be an epithelioid sarcoma. CASE REPORT: A 78-year-old woman had an indolent ulceration of the left sole for several months. Physical examination disclosed a well demarcated 3-cm ulcerated lesion with a red center and flat edges. Skin sections confirmed the diagnosis of epithelioid sarcoma. Cells stained positively for anti-vimentin, anti-cytokeratin and anti-epithelial membrane antigen, but not for anti-S100 protein and anti actin antibodies. Wide local excision was performed. DISCUSSION: Epithelioid sarcoma is an uncommon malignant tumor which apparently differentiates from mesenchymatous cells. It is usually observed in the distal extremities in young males, predominantly the hands and forearms. The tumor presents a firm, flesh colored indolent nodule. Ulceration usually develops and involves the subcutis and deeper soft tissue, particularly fascial planes, aponeuroses and tendon sheaths. Treatment is wide surgical excision with or without radiotherapy. The case reported here on the sole of the foot in a 78-year-old women is unusual. Clinicians should be aware that the initial biopsy may not be contributive and that repeated biopsies may be necessary for positive diagnosis. PMID- 9747244 TI - [Cutaneous sarcoidosis secondary to inhalation of wall insulating material particles]. AB - BACKGROUND: Sarcoidosis is a systemic disease defined by multiple granulomas. We report a case of sarcoidosis which occur concomitantly or secondary to foreign body granuloma of the lung. CASE REPORT: A 50-year-old women presented with Lofgren syndrome, subcutaneous granulomatous nodular lesions on the arms and legs. Computed tomography revealed a foreign body granuloma of the lung centered on particles of mural isolation material that the patient had inhaled accidentally. Analysis of the foreign body particles showed non crystalline silica, calcite monohydrate and phenol resin. DISCUSSION: Recent studies support the hypothesis that sarcoidosis is an antigen-driven disease involving pulmonary T-cell activation. The antigen-mediated reaction may be caused by infectious agents, particularly mycobacteria, occupational and environmental agents (beryllium) or as in our observation following inhalation of mural isolation material particles. PMID- 9747245 TI - [Pheochromocytoma manifesting as toe necrosis]. AB - BACKGROUND: Cutaneous manifestations of pheochromocytoma other than sweating, and facial pallor during paroxysmal episodes of hypertension are exceptional. CASE REPORT: We observed partial necrosis of the fourth toes which revealed pheochromocytoma. DISCUSSIONS: Signs of peripheral vascular disease are uncommon during the course of pheochromocytoma. Only four cases have been reported in the literature. Occurrence of distal necrosis in combination with hypertension and palpable pulses is suggestive of pheochromocytoma requiring assay of urinary catecholamines. The pathogenic mechanisms of necrosis would be vasospasm of cutaneous vessels due to excessive plasmatic catecholamine levels and thrombocytosis as an aggravating factor. PMID- 9747246 TI - [Bullous erythema nodosum leprosum. A case report in French Guiana]. AB - BACKGROUND: Polar and borderline lepromatosis leprosy can be complicated by type 2 reactional states, including erythema nodosum leprosum. CASE REPORT: A 36-year old man was treated for lepromatous leprosum. He consulted for recurrent erythema nodosum leprosum. Certain nodular lesions were bullous. Histopathology demonstrated major dermal edema causing bullous blisters. Treatment with thalidomide led to rapid regression. DISCUSSION: The bullous form of erythema nodosum leprosum is rarely described in the literature. It raises the diffential diagnosis with other bullous dermatoses, particularly Sweet's syndrome. PMID- 9747247 TI - [Superior vena cava thrombosis manifesting as vasomotor facial flushes]. AB - BACKGROUND: The first sign in the reported case of superior vena cava thrombosis secondary to a pacemaker lead, was exceptional: facial flush. CASE REPORT: A 52 year-old woman had a pacemaker for 10 years for rhythm disorders. She developed facial flush triggered by exercise and anteflexion. The clinical examination revealed collateral thoracic circulation, suggesting thrombosis of the superior vena cava which was confirmed by the angiocavogram. DISCUSSION: Vasomotor flush is an uncommon and misleading initial sign of superior vena cava thrombosis. Induction by exercise and anteflexion is characteristic. Due to the increasing number of implanted patients, clinicians should be aware that pacemaker leads are an uncommon cause of superior vena cava thrombosis. PMID- 9747248 TI - [Pachydermoperiostosis with extramedullary hematopoiesis without myelofibrosis]. AB - BACKGROUND: In three previous reports of primary hypertrophic osteoarthropathy, an associated extramedullary hematopoiesis was related to myelofibrosis. CASE REPORT: A 44-year-old male patient with primary hypertrophic osteoarthropathy diagnosed when he was 34-year-old was referred to our hospital with an abdominal mass fortuitously detected. DISCUSSION: The present case is unique for the patient developed an extramedullary hematopoiesis without associated myelofibrosis. It suggests the possible intervention of growth factors common to the skin fibroblasts and the blood progenitor cells in the pathogenesis of primary osteoarthropathy. PMID- 9747249 TI - [Cutaneous manifestations of erysipeloid septicemia]. AB - BACKGROUND: Rouget du porc, or swine erysipelas, usually occurs in man as Rosenbach's erysipeloid. Septicemic forms are more uncommon and can be associated with dermal involvement far from the site of inoculation. We report a case in a patient given corticosteroid therapy for systematic lupus. CASE REPORT: A 50-year old farmer was seen with fever, infiltrative erythema of the long finger and dorsal lesions on the ring finger which developed after a skin lesions caused by a duck. The diagnosis of septicemic rouget du porc was made after isolating the germ from blood cultures. There was no associated endocarditis. Fever and skin lesions totally regressed after treatment with ceftriaxone. DISCUSSION: The diagnosis of erysipeloid was supported by epidemiologic arguments and characteristic clinical features. The corticosteroid therapy was probably a favoring factor for development of septicemia. Positive diagnosis is usually obtained from blood culture but the germ can be isolated from skin biopsies at the site of inoculation. Our patient was free of endocarditis which should always be suspected. Endocarditis is frequent and often fatal. Intravenous high-dose penicillin G is recommanded treatment. PMID- 9747251 TI - [Urticaria and anaphylactic shock from benfluorex. Two case reports]. PMID- 9747250 TI - ["Blueberry muffin baby"]. AB - BACKGROUND: Blueberry muffin baby is a characteristic neonatal syndrome characterized by multiple dark-bluish skin nodules. The clinical significance and prognosis of this syndrome are variable. CASE REPORT: A male child was born to non-consanguinous parents. At birth, a polymalformative syndrome associated macrostomy, bilateral cryptochidy and hexadactyly. There were also about twenty firm dark-bluish skin nodules disseminated over the entire body. These skin lesions regressed spontaneously within one month. Pathology examination of a skin nodule showed lymphomonocyte proliferation. Immunostaining favored T cell infiltration without monoclonal proliferation. Medullar genome mapping showed evidence of a fragile site on the end of chromosome 20. At 8 months the child had normal development. DISCUSSION: We attributed this blueberry muffin baby syndrome to T cell proliferation but we were unable to distinguish between extramedullary leukopoiesis and leukemia. Despite the absence of systematic disease and the complete regression, no exact diagnosis and prognosis could be established in the case. The association of blueberry muffin baby syndrome with a polymalformative syndrome was probably related to a genetic anomaly on chromosome 20 not previously reported. PMID- 9747252 TI - [Coronary disease during subacute lupus erythematosus]. PMID- 9747253 TI - [Case for diagnosis. Fabry's disease]. PMID- 9747254 TI - [Case for diagnosis. Actinic granuloma]. PMID- 9747255 TI - [Diagnosis of "fish odor syndrome" by urine nuclear magnetic resonance proton spectrometry]. AB - BACKGROUND: Trimethylaminuia is an unusual observation, often termed fish odor syndrome. The condition results from reduced ability to oxidize trimethylamine (TMA), which has a fishy odor, into odorless trimethylamine N-oxide (TMAO). METHOD: Proton nuclear magnetic resonance spectroscopy (MRS) was used as a simple and rapid method to detect TMA and TMAO in the same urine sample without pretreatment. Subjects were considered to have deficient N-oxidation of TMA if the TMAO/TMA ratio was greater than 80 p. 100 (heterozygous) or 65 p. 100 (homozygous). DISCUSSION: Direct proton RRS analysis of urine is well suited for diagnosis of fish odor syndrome. It can be used to detect heterozygous patients and also provides an easily implemented follow-up tool. PMID- 9747256 TI - [Management of severe atopic dermatitis]. PMID- 9747258 TI - [Is delayed diagnosis still the main prognostic factor in melanoma?]. PMID- 9747257 TI - [Palmoplantar psoriasis: diagnosis and therapeutic strategy]. PMID- 9747259 TI - [Melanoma in the Strasbourg University Hospital. A 30-year study]. AB - INTRODUCTION: The incidence of melanoma has increased more than the incidence of any other cancer in the past twenty years. There is no cure for advanced-staged melanoma and it's early diagnosis has become a main issue of health policy. Therefore, precise epidemiological data are needed. These data depend on the geographical setting and very few data are available for France. We studied these data in Strasbourg. PATIENTS AND METHODS: All patients hospitalized for melanoma between 1960 and 1989 at the academic dermatology department, Strasbourg, were included in this retrospective, monocentric, study. The diagnosis of melanoma was confirmed in all cases by a dermatopathologist. Clinical, histopathologic and epidemiological data of the patients were recorded. RESULTS: Six hundred seventeen patients with a mean age of 52 years were included. The number of new patients hospitalized for melanoma grew steadily. The mean tumor thickness was 2.31 mm (+/- 1.59) and it remained unchanged between 1970 and 1989. The mean duration between the first clinical signs of melanoma and excision of the tumor was 22 months. Only 4 p. 100 of the melanomas were diagnosed by means of routine examination, and this concerned almost exclusively patients hospitalized after 1980. Eighty five p. 100 of the patients had localized melanoma (stage I) at time of diagnosis. Forty two p. 100 of the patients developed metastasis. The mean five year survival rate was 68 p. 100. CONCLUSION: The number of new patients hospitalized each year for melanoma grew steadily, but the mean tumor thickness remained unchanged. This indicates that the ratio "thick"/"thin" melanomas has remained unchanged between 1960 and 1989 and that the number of melanomas of any thickness has increased. Clinical data show an unawareness of the local population of the dangers of pigmented lesions during the reference period. This unawareness can be partially explained by the fact that no specific information campaign has ever taken place in this area. These data suggest that such a campaign should be recommended. PMID- 9747260 TI - [Pachydermodactyly: seven new cases]. AB - INTRODUCTION: Pachydermodactyly is a superficial benign digital fibromatosis usually involving the proximal portions of the fingers. It is clinically characterized by an asymptomatic, bulbous, soft-tissue swelling around proximal phalanges and interphalangeal joints. We report here seven new cases of pachydermodactyly. CASE REPORTS: Seven patients (4 F, 3 M) ranging in age from 14 to 63 years were studied. Two of them were affected by tuberous sclerosis; two other patients were sisters, one of whom was affected by the transgrediens form of pachydermodactyly. The personal history of two male patients revealed the compulsive habit of interlacing the fingers. Finally a 23-year-old patient was affected by the localized pachydermodactyly. In all the patients roentgenogram and echography of the affected fingers as well as histological and ultrastructural of a cutaneous biopsy examination were carried out. The results of the tests confirmed the diagnosis of pachydermodactyly. DISCUSSION: Our data suggest that pachydermodactyly is underestimated rather than rare and more frequent in females than in males as until now reported in the literature. We suggest classifying pachydermodactyly into five types: classic pachydermodactyly frequently associated with mechanical trauma, monopachydermodactyly or localized pachydermodactyly, transgrediens pachydermodactyly in which the cutaneous thickness extends to the metacarpophalangeal areas, familial pachydermodactyly which may by transgrediens and pachydermodactyly associated with tuberous sclerosis. PMID- 9747261 TI - [Orbital edema induced by psychotropic drugs?]. AB - INTRODUCTION: Drugs known to be responsible for orbital edema are calcium channel blockers, rifampicin, atracurarium, methylprednisolone and mannitol. We describe three patients with orbital edema induced by psychotropic drugs administrated for a long period of time. OBSERVATIONS: These patients were aged 51 to 73 years, and had orbital edema predominant in the morning, but persisting during the day. All the other causes of orbital edema--including steatoblepharon--were ruled out by medical history and investigations. In one case the edema decreased after the interruption of nearly all psychotropic drugs. DISCUSSION: The exclusive orbital localization of the edema suggests a local or regional mechanism that could be an exacerbation of the physiological morning orbital edema induced by decreased lymphatic flow. This flow is usually enhanced by gravity and automatic palpebral blinking. PMID- 9747262 TI - [Giant cell fibroblastoma associated with Darier-Ferrand dermatofibrosarcoma in an adult]. AB - INTRODUCTION: Giant cell fibroblastoma is a rare mesenchyma tumor of childhood having many similarities with dermatofibrosarcoma protuberans in adults. OBSERVATION: We report the case of a 28-year-old woman presenting a subcutaneous inter-mammary mass associating both tumors. Immunohistochemistry showed an expression of CD 34 only by dermatofibrosarcoma protuberans cells. DISCUSSION: It is important to stress: the rarity of this association, the difficulty to confirm the diagnosis and to establish the links between these two tumors: simple association (as in our case), transformation or recurrence of giant cell fibroblastoma in dermatofibrosarcoma protuberans. PMID- 9747263 TI - [Kasabach-Merritt syndrome on a congenital tufted angioma]. AB - INTRODUCTION: Kasabach-Merritt syndrome is a very rare disease of infancy, with profound thrombocytopenia and a mild to severe consumption coagulopathy; this biological phenomenon is difficult to control. CASE REPORT: A 1-month old boy had a congenital plaque-like lesion in the calf. It was a biopsy-proven tufted angioma. Five weeks later, Kasabach-Merritt syndrome developed. After failure of ticlopidine + aspirin, and oral betamethasone treatment, thrombocytopenia was cured with vincristine treatment, then the leg lesion slowly continued to shrink after cessation of the treatment. It had disappeared before the age of 1 year. DISCUSSION: We highlighted two points: 1) Kasabach Merritt does not appear as a complication of a classic hemangioma (infantile, "cellular", "capillary", involuting-type), as it has long been thought. In our experience, it develops on a different endothelial cell proliferation, in this case a congenital tufted angioma, but it can also engraft on a kaposiform hemangioendothelioma. 2) These patients are difficult to treat because, up to now, no single treatment has given constant by good results. Vincristine was recently introduced in the treatment of Kasabach-Merritt syndrome, with excellent, rapid outcome. CONCLUSION: What seems a therapeutic progress in a difficult field needs further control. PMID- 9747264 TI - [Seborrheic keratosis erupting in a tattoo]. AB - INTRODUCTION: Decorative tattoos have been associated with inflammatory reactions and transmission of infectious diseases. Cutaneous tumors have rarely been reported. CASE REPORT: We report the case of a 26-year-old man who presented eruptive seborrheic keratoses strictly localized on the area of a decorative tattoo. No other lesion was present anywhere else on the cutaneous surface. Three years later the lesions remained stable. COMMENTS: To our knowledge, this is the first report of eruptive seborrhelc keratoses on a tattoo. In our observation, the role of human papillomavirus contamination during tattoo procedure is discussed. PMID- 9747266 TI - [Disseminated cutaneous leishmaniasis revealing human immunodeficiency virus infection]. AB - BACKGROUND: Unlike visceral leishmaniasis, cutaneous leishmaniasis is uncommonly described in patients with human immunodeficiency virus infection. Diffuse cutaneous forms due to Leishmania infantum are always accompanied by visceral parasite infection. CASE REPORT: We report a case of cutaneous leishmaniasis without visceral extension which was the inaugural sign of immunodeficiency virus infection. Polymerase chain reaction amplification of the genome was used to identify Leishmania infantum. Pentamidine (2 injections at the dose of 4 mg/kg, separated by one week) followed by maintenance therapy at the same dose every two weeks was given. Clinical cure was obtained after the initial injections and the intracellular parasite infestation had disappeared at the histology control. DISCUSSION: This case was unusual in that it was the inaugural sign of HIV infection. In addition, it is the first reported case of diffuse cutaneous leishmaniasis without visceral extension of Leishmania infantum in an HIV infected patient. As no therapeutic consensus has been established, pentamidine would appear to be interesting for this population which also requires pneumocystosis prophylaxy. PMID- 9747265 TI - [Epidermolysis bullosa acquisita and hepatitis C]. AB - INTRODUCTION: Epidermolysis bullosa acquisita is a bullous dermatosis. Its etiology remains unknown and the efficacy of its treatment is low. OBSERVATION: We report the first association between epidermolysis bullosa acquisita, chronic hepatitis C and cryoglobulinemia, healing with interferon alpha and ribavirine. DISCUSSION: We suggest a role for hepatitis C virus in the pathogenesis of epidermolysis bullosa acquisita. We suppose a synthesis of autoimmune antibodies in a dysimmune environment. Interferon alpha and ribavirine might be a new therapeutic avenue but further studies are necessary to confirm it. PMID- 9747267 TI - [Hair follicles and vascular endothelial growth factor]. PMID- 9747269 TI - [Legality in the prescription of dermatologic agents]. PMID- 9747268 TI - [Maxilene and alopecia]. PMID- 9747270 TI - [Case for diagnosis. Blue nevus]. PMID- 9747271 TI - [Case for diagnosis. Reiter syndrome]. PMID- 9747272 TI - [Pigmented eccrine poroma]. PMID- 9747273 TI - [Morphea: classification and management]. PMID- 9747274 TI - [Sensitivity and specificity revisited: significance of the terms in analytic and diagnostic language]. AB - Imprecise usage of the terms "sensitivity" and "specificity" produces confusion in the diagnostic use sophisticated laboratory test results. "Analytical sensitivity" represents the smallest amount of substance in a sample that can accurately be measured by an assay. "Analytical specificity refers to the ability of an assay to measure one particular organism or substance, rather than others, in a sample. An assay's analytical sensitivity and analytical specificity are distinct from that assay's clinical diagnostic sensitivity and diagnostic specificity. Diagnostic "sensitivity" is the percentage of persons who have a given disorder who are identified by the assay as positive for the disorder. High analytical sensitivity does not guarantee acceptable diagnostic sensitivity. "Diagnostic sensitivity" is the percentage of persons who do not have a given condition who are identified by the assay as negative for the condition. False positive reactions occur because of sample contamination and diminish the diagnostic specificity of the assay. The terms "sensitivity" and "specificity" should be used with the requisite adjectives because the "diagnostic" and the "analytical" meanings of these terms are very different. PMID- 9747275 TI - [The monthly question. Is the systematic use of standard set of allergens lawful?]. PMID- 9747276 TI - [Drug hypersensitivity syndrome: research of etiology]. PMID- 9747277 TI - [Importance of bFGF ("basic fibroblast growth factor") for diagnosis and treatment of hemangiomas]. AB - OBJECTIVES: Hemangiomas of infancy follow a characteristic three-phases course: proliferation, involution, regressed Proliferative endothelial cells predominate during the proliferative phase. Moreover it has been shown that patients with active angiogenesis have elevated levels of urinary bFGF (basic Fibroblast Growth Factor). PATIENTS AND METHODS: Here we report our preliminary results of urinary bFGF assay (ELISA) for the diagnosis and follow up of severe hemangioma. We also assayed bFGF in normal infants, in patients with large vascular malformations and in infants with Kasabach-Merritt syndrome. RESULTS: In the control group, urinary bFGF was elevated in new borns but nul or very low in infants. Urinary bFGF levels were normal, i.e. very low in 4 patients with a vascular malformation. In infants with a clinically proliferative hemangioma, urinary bFGF was elevated in 8 among the 10 studied. bFGF levels guided treatment in 9 patients. Urinary bFGF was elevated in 4 patients with Kasabach-Merritt syndrome. DISCUSSION: Angiogenesis is regulated by angiogenic and inhibitory factors. The angiogenic factor bFGF is an autocrine growth factor for endothelial cells and hemangioma endothelial cells expressing bFGF in their cytosol during the proliferative phase. As suggested by J. Folkman and his group, assay of urinary bFGF appears useful in differentiating between hemangioma and vascular malformation and for follow up of treated patients. PMID- 9747278 TI - [Dorfman-Chanarin syndrome]. AB - BACKGROUND: Dorfman-Chanarin syndrome is an uncommon condition characterized by non-bullous congenital ichtyosiform erythrodermia and lipid vacuoles in circulating leukocytes. CASE REPORT: We report an unusual presentation in a child who had a dry congenital ichtyosiform erythroderma. Blood smears revealed lipid vacuoles in granulocyte cytoplasm, leading to the diagnosis of Dorfman-Chanarin syndrome. The child also had liver and ophthalmologic involvement. DISCUSSION: Dorfman-Chanarin syndrome is a rare autosomic recessive hereditary disease (27 cases reported in the literature) related to the accumulation of neutral lipids in organ tissues. Clinical manifestations are dry congenital ichtyosiform erythroderma and lipid vacuoles in circulating granulocytes. The syndrome may be expressed more or less severely in several organs. Diagnosis is confirmed on blood smears. The vacuoles can also be observed in smears of heterozygous subjects and can serve as a screening test. The pathogenesis of Dorfman-Chanarin syndrome is poorly understood but appears to be related to perturbed intracellular triglyceride catabolism. Treatment is symptomatic. PMID- 9747279 TI - [Epidermolysis bullosa acquisita and metastatic cancer of the uterine cervix]. AB - INTRODUCTION: Epidermolysis bullosa acquisita may be associated to a systemic or malignant disease. Here, we report a case of epidermolysis bullosa acquisita associated with a carcinoma of the cervix. CASE REPORT: A 44-year-old woman presented inflammatory, bullous and erosive mucocutaneous lesions. Investigations lead to the diagnosis of epidermolysis bullosa acquisita and revealed pelvic metastases originating from a poorly differentiated carcinoma. The cutaneous lesions completely regressed after the treatment of the tumor but reappeared with tumoral relapse. DISCUSSION: This is the first report of an association of epidermolysis bullosa acquisita and carcinoma of the cervix. The clinical course of these two entities suggests that the EBA in this case may be paraneoplastic. PMID- 9747280 TI - [Multiple and unilateral angiofibromas of the face: forme fruste of Bourneville tuberous sclerosis]. AB - BACKGROUND: Multiple facial angiofibromas are considered a characteristic and pathognomonic feature of tuberous sclerosis. In contrast, the observation of localized multiple angiofibromas limited to one side of the face is a uncommon. OBSERVATIONS: We cared for two patients with multiple clustered unilateral angiofibromas (although one patient also presented a few angiofibromas on the other side) without other features associated with the sclerosis tuberous complex. DISCUSSION: The possibility that these patients presented an independent disorder or a minor form of tuberous sclerosis is discussed. PMID- 9747281 TI - [Hypersensitivity syndrome during Mycoplasma pneumoniae infection]. AB - BACKGROUND: Skin manifestations have been described in 25% of patients with Mycoplasma pneumoniae infection. CASE REPORT: We report a case of Mycoplasma pneumoniae infection in a 29-year-old man who developed a polymorphous erythema like reaction. Skin manifestations were associated with voluminous lymph node enlargement and high eosinophil levels both in serum and alveolar lavage. Seroconversion against Mycoplasma pneumoniae IgG was documented with ELISA. The clinical course was favorable with erythromycin. DISCUSSION: ELISA IgG seroconversion is sufficient to confirm the diagnosis of Mycoplasma pneumoniae infection as this test has an 80-90% specificity. This case was unusual by its clinical presentation and high eosinophil counts in serum and tissue samples, similar to what is found in drug-induced hypersensitivity. PMID- 9747282 TI - [Pyostomatitis-pyodermatitis vegetans uncovering a case of Crohn disease]. AB - BACKGROUND: Pyostomatitis-pyodermatitis vegetans is an uncommon condition associated with chronic inflammatory bowel disease in 75% of the cases, usually hemorrhagic rectocolitis. CASE REPORT: A 48-year-old man was referred for recent development of pustulous lesions of the lips and buccal mucosa and weight loss. He complained of abdominal pain and intermittent diarrhea which had persisted for more than one year. During the last three months, a pseudotumoral plaque with a pustulous rim had developed over the two distal phalanxes of the right middle finger in association with ungueal lysis and nodular, vegetating and crusted lesions on the lateral aspect of the left arm. Small pustules covered the entire buccal mucosa excepting the tongue and the glans forming a typical snail trace aspect. Bacterial and mycological samples were negative. The histology reports for skin and mucosa were similar: epithelial hyperplasia, intra- and subepithelial granulocyte micro-abscesses and polymorphous infiltration of the superficial derma with numerous neutrophils and eosinophils. There was a discrete interkeratinocytic fluorescence at direct immunofluorescence but indirect immunofluorescence was negative. Anti-desmogleine 1 immunolabeling showed typical normal skin uptake and immunotransfer was negative. Digestive tract endoscopy and histopathology examination of the bowel specimens confirmed the diagnosis of Crohn's disease. Clinical manifestations improved dramatically with prednisone. DISCUSSION: This case of pyostomatitis-pyodermatitis vegetans involved several aspects rarely reported in the literature: a) the cutaneomucosal signs were inaugural; b) the association with Crohn's disease; c) the presence of lesions to the genital mucosa; d) the unusual localization of the inaugural skin manifestations. This clinical entity has now been clearly distinguished from pemphigus vegetans. There was however a long debate on the similar clinical, histological and even immunological expressions. We suggest that pyostomatitis pyodermatitis vegetans belongs to the spectrum of neutrophilic dermatoses and other authors even propose it is a clinical form of pyoderma gangrenosum. PMID- 9747283 TI - [Langerhans-cell histiocytosis and Erdheim-Chester disease: probably not a fortuitous association]. AB - BACKGROUND: Erdheim Chester disease (MEC) is a rare non-Langerhans cell histiocytosis characterized by multi-visceral involvement. We report a case of MEC associated with Langerhans cell histiocytosis (HCL). CASE REPORT: A 46-year old women presented skin and vulvar localization of HCL associated with typical MEC bone involvement. Despite chemotherapy (vinblastine) and prednisone, the disease progressed to involve the central nervous system, leading to fatal outcome. Post-mortem examination showed HCL in skin, MEC in bones and central nervous system, and intermediate histiocytic proliferation in the encephalon. DISCUSSION: Usually, MEC and HCL are considered as distinct entities. MEC is characterized by a xanthogranulomatous proliferation of CD 68+ foamy histiocytes nested in fibrosis, and HCL by a proliferation of PS 100+ and CD1a+ Langerhans cells. However, our observation, as well as previous reports, suggests that MEC is part of the HCL spectrum. PMID- 9747284 TI - [Male predominance of leg ulcer in Morocco]. PMID- 9747285 TI - [Risk of viral infection and the CO2 laser]. PMID- 9747286 TI - [What is a hamartoma?]. PMID- 9747287 TI - [Diagnostic case. Angiotrophic lymphoma with T-cell immunophenotype]. PMID- 9747288 TI - [Diagnostic case. Keratoderma of the palm creases]. PMID- 9747290 TI - [Contribution of immunohistochemistry to diagnosis of cutaneous tumors]. PMID- 9747289 TI - [Compression therapy of chronic venous insufficiency]. PMID- 9747291 TI - [Question of the month. Should sunscreening agents be evaluated like drugs?]. PMID- 9747292 TI - [Disease associations: risk of necessity?]. PMID- 9747293 TI - ["Hepatitis, the vaccine, multiple sclerosis and lichen": the story and its morale]. PMID- 9747294 TI - [Lichen planus in children: role of the campaign for hepatitis B vaccination]. AB - INTRODUCTION: Cutaneous lichen planus in children is a rare entity. To study its specific clinical and therapeutic features, we report on 8 cases seen in 5 years in a pediatric dermatology clinic. In parallel, this study addresses the role of HBV immunization campaigns in childhood lichen planus. OBSERVATIONS: Six boys and two girls with a diffuse form of lichen planus were studied. The mean age was 11 years. Four children were of skin types V and VI. Lesions followed Blaschko's lines in two children. One case was associated with atopic dermatitis. In half the cases--the most recent--the eruption was noted after HBV immunization. Biopsy specimens from seven children were reviewed and confirmed the diagnostic of lichen planus. Oral retinoids or corticosteroids were used successfully in six cases. DISCUSSION: Half of our cases confirmed the atypical features of lichen planus in childhood. This study confirms the predominance of lichen planus in children with pigmented skin suggesting a genetic predisposition. The eruption following Blaschko's lines suggests that a clonal keratinocytic population is the target of lichenoid inflammation. HBV immunization could be a stimulus triggering a cytotoxic lymphocyte-mediated reaction. The proportion of affected children was low in comparison with all children immunized. So lichen planus arises probably in predisposed children. This justifies surveillance in the pediatric and dermatologic medical communities. PMID- 9747295 TI - [Chronic radiodermatitis after interventional cardiac catheterization. Four cases]. AB - BACKGROUND: Fluoroscopic and cineradiographic procedures expose patients undergoing coronarography to high doses of ionizing irradiation. CASE REPORT: We report four cases of radiodermitis following cardiac catheterization. A 69-year old man developed a radio-induced ulceration on the left scapular region in 1991 which required excision with skin graft. He had undergone 3 coronarographies and 2 angioplasties from 1989 to 1991. In 1992, a 59-year-old women developed a hard dorsal lesion with central ulceration and scar formation requiring excision and graft. From 1990 to 1992, she had undergone two coronary dilatations with angioplasty during one procedure. An atrophic necrotic wound situated under the right nipple developed in a 63 year old man. Excision with flap reconstruction was performed in 1993, two years after an unsuccessful angioplasty then two vessel bypass. In a fourth case, a 52 year-old woman developed a telangiectasic ulceration on the right breast in 1990. The diagnosis of radio-induced dermitis was confirmed in 1996 and the patient was treated by excision. She had had three angioplasties in 1989. DISCUSSION: Four other cases of radio-induced dermatitis following cardiac catheterism have been reported in the literature since 1996. Six other cases were also recently reported in France. All of these patients had undergone coronarography with transluminal coronary angioplasty. Besides coronarography, irradiation exposure is greatest for guide and balloon insertion required for dilatation procedures. Angioplasty is particularly dangerous because the irradiation beam is focused on the stenosis while the entire coronary network is concerned for coronarography. In most cases of radio-induced dermatitis following cardiac catheterism, the diagnosis is usually evident from the clinical context and the localization of the coronary lesion. In many cases however, the long delay to onset may make diagnosis a difficult task. In addition, the radiation dose delivered to the skin during cardiac procedures is not measured. PMID- 9747296 TI - [Actinic lichen planus: 32 cases]. AB - OBJECTIVES: Actinic lichen planus is a variant form of lichen planus located on light-exposed areas, generally in children or young adults with dark skin living in tropical countries. Three forms have been described: annular, pigmented and dyschromic forms. The aim of this work was to determine the epidemiological, anatomic and clinical features of actinic lichen planus in our region. PATIENTS AND METHODS: A retrospective study included 32 patients with actinic lichen planus on the basis of clinical features and histological findings. The geographical origin, age, sex, phototype and clinical and histological characteristics were recorded for each case. RESULTS: The incidence of actinic lichen planus was estimated at 1 case per million inhabitants per year. Age in our patients ranged from 7 to 47 years (mean = 17 years), with female predominance (F/M = 2.5). Twenty-six patients were from the Sfax area. Phototypes ranged from III to V, mainly type IV (43.7%). Onset was usually in spring (in 18 cases). Facial involvement was the most frequent (91%) and the annular form predominated (84%); the pigmented form was found in 4 patients and the dyschromic form in 1. Histology showed typical actinic lichen planus in 18 cases and lichen planus in 14 cases. DISCUSSION: Compared with data reported in the literature, our series showed a low incidence, younger patients and greater female predominance. The annular form appeared to be the most specific. The pigmented form raises problems of differential diagnosis with melasma. Certain histological signs are highly suggestive. PMID- 9747297 TI - [Faun tail and sacral hemangioma associated with occult spinal dysraphism]. AB - INTRODUCTION: Hemangiomas are the most frequent skin tumor of childhood. Usually, a "wait and see" policy is adopted. However, a sacral hemangioma may reveal occult neurodysraphism. CASE REPORT: MRI discovered lipomyelomeningocele and a tethered spinal chord in an asymptomatic 4-month-old boy with sacral hemangioma and faun tail. Because of absence of neurological defect only surgery of the caudal appendage was performed for the moment. DISCUSSION: Midline lombosacral lesions, as well as lipomas, hirsutism or pilonidal cyst, may be associated with occult spinal defect; the most severe is tethered chord. Renal or ano-genital anomalies can be also associated. CONCLUSION: MRI is necessary in case of midline sacral hemangioma to detect underlying anomalies. PMID- 9747298 TI - [Infiltrating lipoma]. AB - BACKGROUND: Deep or infiltrating lipoma is an often misdiagnosed clinical entity. We report two typical cases. CASE REPORTS: Two women, aged 92 and 62 years, were seen for a tumefaction on an upper limb with progressively increasing volume. The clinical presentation suggested deep lipoma, confirmed by magnetic resonance imaging and histology which eliminated liposarcoma, the main differential diagnosis. Surgical excision was successful. There has been no recurrence. DISCUSSION: These two observations recall the clinical, diagnostic and therapeutic features of infiltrating lipoma. PMID- 9747299 TI - [Neutrophilic eccrine hidradenitis associated with relapse of acute myeloblastic leukemia]. AB - BACKGROUND: Neutrophilic eccrine hidradentitis is a recently described clinical entity. Most reported cases have occurred in patients given chemotherapy for acute myelogenous leukemia, suggesting a drug induced mechanism. Some authors have considered however that neutrophilic eccrine hidradenitis belongs to the group of neutrophilic dermatoses. CASE REPORT: We observed neutrophilic eccrine hidradentitis in a 48-year-old man which developed when he suffered a relapse of acute leukemia. He had not been given chemotherapy in the preceding months. DISCUSSION: This case favors the hypothesis that neutrophilic eccrine hidradenitis is associated with myeloid hemotology disorders and not a complications secondary to treatment. PMID- 9747300 TI - [Endemic subcutaneous Dirofilaria repens filariasis in Southern France]. AB - BACKGROUND: Human dirofilariosis is endemic in southern France. The causal agent is Dirofilaria repens transmitted to man by mosquitos. The human organism is an accidental host while the dog is the reservoir. Approximately 60 cases have been reported in France, mainly in southern continental areas and Corsica. Most cases involve subcutaneous and occular manifestations. CASE REPORT: A 47-year old woman living in the Var department in southern france consulted for a subcutaneous nodule with the aspect of an epidermoid cyst. At incision, the nodule was found to contain a helminth found at parasite examination to be Dirofilaria repens. DISCUSSION: The prevalence of endemic subcutaneous dirofilariosis is probably underestimated as the clinical expression is non-specific and spontaneous cure is common. The diagnosis of epidermoid cyst is frequently suggested. History taking does not reveal travel to intertropical areas. The helminth is discovered when the nodule is opened, also providing successful cure. PMID- 9747301 TI - [Membranous lipodystrophy caused by chemotherapy]. AB - INTRODUCTION: Membranous lipodystrophy represents a peculiar dermatopathologic type of cystic forming adipose tissue necrosis. In skin pathology two distinct entities are currently known: a primary idiopathic type and a secondary type found in association with various cutaneous or systemic diseases (lupus erythematosus, scleroderma, dermatomyositis, venous disorders, trauma, diabetes mellitus...). This secondary type is more common. CASE REPORT: We report the case of a female patient treated with cytostatic regimen for metastatic adenocarcinoma of the breast. She was seen with cyclic painful attacks of hypodermitis of lower limbs following drug infusions. Attacks finally vanished after treatment withdrawal. Membranous lipodystrophy was observed microscopically. DISCUSSION: This is, to the best of our knowledge, the first case of membranous lipodystrophy occurring during cytostatic treatment. The origin of such a phenomenon remains nuclear but cystic formation occurs after adipocyte necrosis and membranes are thought to be formed from dead cells plasma membrane remnants. Primary cell injury could be due to ischemia to which adipose tissue is especially susceptible. In our case, role of circulatory troubles or cytostatic chemotherapy could be equally discussed. However cyclic attack of painful hypodermatitis following drug administration was strongly in favour of treatment responsibility. Moreover clearing of lesions after treatment discontinuation brought more arguments for this last hypothesis. PMID- 9747302 TI - [Dermatomyositis and polymyositis in Lome (Togo)]. PMID- 9747303 TI - [Acne, hyperandrogenism and resistance to oral isotretinoin: 23 cases]. PMID- 9747304 TI - [Case for diagnosis. Chronic atrophic polychondritis]. PMID- 9747306 TI - [Ocular manifestations of retinoids]. PMID- 9747305 TI - [Case for diagnosis. Toxocariasis]. PMID- 9747307 TI - [Treatment of warts]. PMID- 9747308 TI - [Management of drug resistant dyshidrotic eczema]. PMID- 9747309 TI - [PubMed]. PMID- 9747310 TI - [Dermatoses caused by estrogens and progesterone: an upcoming nosology]. PMID- 9747311 TI - [Neurofibromatosis: pat and prospects]. PMID- 9747312 TI - [Poisonous caterpillars in French Guyana. 5 cases]. AB - BACKGROUND: Certain caterpillars produce venomous substances and cases of human envenomation are regularly published. CASE REPORTS: We report 5 cases of caterpillar-induced envenomation observed in French Guyana. The caterpillar bites produced variable clinical manifestations. Automeris liberia provoked acute pain and skin necrosis: Dirphia tarquinia, erythema; Hylesia, persistent erythematous plaque (4 days); Megalopyge, erythematous and edematous lesions at the site of the bite and distant skin lesions; Automeris, syncopal pain and edematous infiltration of the thigh lasting several days. DISCUSSION: Caterpillar envenomation necessitates consultation, emergency unit care, or even hospitalization. It is important to identify the causal caterpillar species in case of envenomation in order to evaluate risk. Lonomia achelous must always be suspected because this species can cause major fibrinogenolysis. PMID- 9747313 TI - [Treatment of mucocutaneous leishmaniasis with pentamidine isothionate]. AB - BACKGROUND: The mucocutaneous form of New World cutaneous leishmaniasis is caused, theoretically, by Leishmania (viannia) braziliensis. Pentad valency antimonic compounds are widely used for treatment of New World cutaneous leishmaniasis. The rate of recurrence is often high when these drugs are used for mucocutaneous forms. Pentamidine has been considered as a second line treatment although little data is available. PATIENTS AND METHODS: Seventeen patients with mucocutaneous leishmaniasis were treated with pentamidine isothionate, 4 mg/kg every 48 hours. A complete oto-rhino-laryngological examination was performed once a week to determine the course of the lesions. Treatment was discontinued when the lesions healed. A follow-up oto-rhinolaryngological examination was also performed after treatment withdrawal and every three months thereafter for an undetermined period. RESULTS: Lesions healed with pentamidine isothionate in 16 patients (94%). Mean dosage required was 2872 mg (2025-4320 mg) for a mean treatment duration of 22 days (12-32 days). Mean follow-up was 13.3 months (3-37 months). Leishmaniasis relapsed in one patient 4 months after treatment withdrawal. DISCUSSION: Pentamidine isothionate is an effective treatment for mucocutaneous leishmaniasis, providing cure with a low rate of recurrence. PMID- 9747314 TI - [Aggravation of acne by isotretinoin. 6 cases, predictive factors]. AB - OBJECTIVE: Acne flare-ups are frequent in the early phase of isotretinoin treatment. Severity varies from one patient to another. Clinical factors favoring a potentially severe course were assessed on the basis of 6 cases. CASE REPORTS: Six male patients, mean age 16.5 years, with inflammatory acne with a major retentional component were studied. Isotretinoin administered at a daily dose 0.5 mg/kg led to explosive development of massive nodulocystic lesions or pyogenic granulomas within two months. The lesions healed at withdrawal of isotretinoin and administration of antibiotics and antiinflammatory drugs. There was important scar sequellae. DISCUSSION: Four concomitant factors were identified which contribute to the development of acne flare-ups: sex (male), young age, retentional form of acne and daily isotretinoin dose 0.5 mg/kg. PMID- 9747315 TI - [High resolution ultrasound imaging: value in treatment of basocellular carcinoma by cryosurgery]. AB - OBJECTIVE: We conducted a prospective evaluation of the contribution of high resolution ultrasound imaging prior to cryosurgery for basocellular carcinoma and in search for recurrence. PATIENTS AND METHODS: All patients seen between 1992 and 1994 at the skin tumor clinic and treated by cryosurgery were included. Ultrasound imaging using 20 MHz prototype was performed prior to cryosurgery and 2 months later. RESULTS: Among 101 patients treated, 112 basocellular carcinomas were treated by cryosurgery. Ultrasound imaging provided good visualization of the tumor limits in all cases. The ultrasound aspect was anechogenic, often with rare areas of highly dense echoes. The tumor limits described by ultrasound imaging were larger than the clinical limits in 32% of the cases. In 8 of the 16 cases of recurrent tumors, the ultrasound examination revealed the recurrence first. In the other 8 cases, clinical manifestations were confirmed by ultrasonography. In our series, recurrence of basocellular carcinoma was statistically more frequent when the depth of the tumor was 3 mm (ultrasonographic measurement) or when the lateral limits established by ultrasound assessment were greater than the clinical evaluation. DISCUSSION: These findings demonstrate that high-resolution ultrasound imaging of basocellular carcinomas prior to cryosurgery: 1) visualizes tumor limits allowing adapted cryosurgery, 2) identifies factors with predictive value for recurrence, 3) can identify recurrences early. Ultrasound imaging of the skin is a useful non invasive technique for pre- and post-therapeutic assessment of skin tumors and could be a particularly useful tool for "blind" cryosurgery destruction of skin tumors. PMID- 9747316 TI - [Dermatitis caused by estrogens]. AB - BACKGROUND: We report a case of sensitization to estrogen. CASE REPORT: A 40-year old woman consulted for skin disorders which followed a cyclic pattern. At each menses, the patient developed pruritus and erythematous papulovesicular lesions over the members and trunk. Estraderm patch contact dermatitis was evident. Prick and patch tests with alcoholic solutions of estrone alone were positive. Serum tests were positive for anti-ethinyl-estradiol antibodies and anti-progesterone antibodies. DISCUSSION: Autoimmune dermatitis can be caused by sensitization to endogenous or exogenous sex hormones. Clinical manifestations and histological findings are variable and non-specific. The cyclic nature of the manifestations is however quite suggestive. Positive prick and patch tests performed with alcohol solutions of the hormones may give the diagnosis and serum tests may be positive for specific anti-steroid antibodies. These complementary explorations are however difficult to perform and interpret and definitive diagnosis is based on an association of clinical findings, skin tests, laboratory tests and the clinical course. In case of progesterone sensitization, the treatment of choice is estrogen inhibition of ovulation. For estrogen sensitization, anti-estrogen treatment appears to be more effective. Finally, bilateral ovariectomy may be required in difficult cases. PMID- 9747317 TI - [Cutaneous-splanchnic neurofibromatosis]. AB - BACKGROUND: Segmental neurofibromatosis (NF V) is ten times less frequent than Recklinghausen disease. Would the risk of visceral involvement in this uncommon form of neurofibromatosis warrant systematic imaging procedures? CASE REPORT: A 31-year-old man consulted for a voluminous plexiform neurofibroma in the left lumbar area. More ventrally, on the left side, there was also a cafe au lait spot. There were no Lisch nodules. The chest and abdominopelvic computed tomography and magnetic resonance imaging showed intramuscular tumoral extension, two neurofibromas in the 9th intercostal space and a voluminous 5-cm tumor situated in the left adrenal area. After resection pathology examination of the surgical specimen confirmed the diagnosis of ganglioneuroma. DISCUSSION: In this patient, all the neurofibromas and the cafe of lait spot developed in the territories of the left T10 and adjacent spinal roots. This was also true for the ganglioneuroma which developed on the deep sympathetic ramus to the adrenal gland which originates essentially from roots T8 to T11. This would place this case in the second subgroup of NF V in Roth's classification. Only six other cases have been reported in the literature. Such deep localizations are very likely to be underestimated, raising the problem of their detection and the correct protocol to follow asymptomatic forms, especially to detect disease progression to malignant degeneration which has a poor prognosis. Patients with a NF V should receive genetic counselling with a search for a family history, other signs of neurofibromatosis and Lisch nodules. In young patients, the risk of deep asymptomatic spread underlines the importance of regional computed tomographic or magnetic resonance explorations. PMID- 9747318 TI - [Maffucci syndrome: a false venous malformation? A case with hemangioendothelioma with fusiform cells]. AB - BACKGROUND: Maffucci syndrome occurs as a sporadic disease. Progressive onset of both cutaneous vascular lesions (considered to be of venous type) and bony enchondromatous tumors (similar to those seen in Ollier disease) occurs throughout childhood. We report a case of a woman with Maffucci syndrome whose cutaneous vascular lesions revealed spindle cell hemangioendothelioma. CASE REPORT: An italian woman developed severe bone distortion and dwarfism due to multiple enchondromas, first diagnosed as Ollier disease during childhood. At puberty, multiple vascular nodules appeared mainly on the limbs, leading to the diagnosis of Maffucci syndrome. Clinical data suggested the diagnosis of cutaneous venous anomalies: blue color of some nodules, phleboliths, arteriographic pattern. Histopathological examination of the skin specimen showed features of spindle cell hemangioendothelioma, e.g. nodules of dense spindle cell infiltration in combination with dysplastic vessels. DISCUSSION: The few reports available do not clearly evidence the underlying histopathology progression of the lesions over years in a given patient. Skin lesions are classified among venous malformations. Bony enchondromatous involvement of the limbs is common and reported in 9 out of 10 patients. Both vascular and bony lesions classically develop from childhood to adulthood. Spindle cell hemangioendothelioma is a vascular tumor recently described. Cellular spindling in association with vascular spaces must not be misdiagnosed as Kaposi sarcoma. Some of the reported cases of spindle cell hemangioendothelioma had Maffucci syndrome. It is unknown whether Maffucci syndrome occurs in association with venous malformation or whether it is always present in the cutaneous vascular lesions of the disease. PMID- 9747319 TI - [Photosensitivity as presenting sign of HIV infection. Control with triple antiretroviral therapy]. AB - BACKGROUND: We report a case of photosensitivity which occurred as the presenting sign of HIV infection. Photosensitivity regressed completely after introducing antiretroviral tritherapy. CASE REPORT: A 44-year-old woman developed a photo distributed eczematous eruption which did not respond to topical steroids or hydroxychloroquine. Histologic examination showed eczematous dermatitis. T-cell marker analysis showed a majority of CD8 cells in the infiltrate. The patient was found to be HIV-positive and CD4 counts were markedly reduced to 190/mm3 while CD8 counts were increased to 1260/mm3. Antiretroviral tritherapy cured the photosensitivity. Cure was apparently related to increased CD4 lymphocyte and normalized CD8 lymphocyte counts. DISCUSSION: Photosensitivity was the presenting disorder of HIV infection in this case. Cure of the photosensitivity with antiretroviral tritherapy has not been reported previously. CD8 T-cell infiltration and very low CD4/CD8 ratio would appear to play a key role in the pathogenesis of photosensitivity in these patients. PMID- 9747321 TI - [Cutaneous malacoplakia: a pediatric case]. AB - INTRODUCTION: Cutaneous malakoplakia is an inflammatory disease characterized by granulomatous accumulation of distinctive phagocytic macrophages. It occurs mainly in visceral or orificial areas; the condition is rarely purely cutaneous, and appears to be extremely rare in childhood. CASE REPORT: A facial cutaneous crusted lesion was diagnosed as cutaneous malakoplakia in an immunocompetent child. The lesion had been excised twice and it had recurred, and the diagnosis was made possible only with a third biopsy, after a 2-year chronic expansion. This third biopsy revealed a dense granulomatous inflammation with numerous phagocytic histiocytes containing abundant fine granules and round Michaelis Gutmann bodies, both staining with PAS, Perls and von Kossa. Biopsy cultures revealed only growth of two different streptococcus (group B) strains. The lesion resolved after a 4-month period of antibiotic therapy, including roxithromycin, ampicillin and trimethoprim-sulfamethoxasole. DISCUSSION: Diagnosis of malakoplakia is mainly made by histopathologic examination of tissue excision or biopsies. There are no specific clinical features. Most reported cases of this uncommon phagocytic reaction to common bacteria have developed in the genitourinary areas (71 p. 100); purely cutaneous localisation, as in our patient, are rare (4 p. 100). Intracytoplasmic granules may result from phagolysosomes and incomplete bacterial killing, with subsequent deposit of iron and calcium in the phagocytic macrophages. A number of reported cases have affected immunocompromised patients with either congenital immunodeficiency or secondary immunodeficiency. The most effective treatment option is based on a protracted antibiotherapy, using drugs that easily permeate the macrophages, e.g. quinolones and trimethoprim-sulfamethoxasole. Lesion may recur after surgical excision. PMID- 9747320 TI - [Pigmented nodular tenosynovitis. A misdiagnosed etiology of isolated tumefaction of fingers or toes]. AB - BACKGROUND: Pigmented nodular tenosynovitis (more commonly called giant cell tumor) is a benign tumor of synovial joint or tendon sheaths. CASE REPORT: A 17 year-old girl had a tumefaction of the first toe. The mass was firm and painless and had been present for one year. Radiological studies revealed a cystic area of the first phalanx. Magnetic resonance imaging demonstrated a poorly vascularized tissular lesion. Surgical excision of the tumor was performed. Macroscopic and histologic findings confirmed the diagnosis of pigmented nodular tenosynovitis. DISCUSSION: Clinical and radiological aspects of nodular tenosynovitis are characteristic. This diagnosis should be made by dermatologists. Eighty percent of cases occur in fingers and more rarely in toes. Patients are usually females (60 p. 100). A subcutaneous mass is the most common presenting sign whereas pain or joint swelling are rare. Optimal treatment is surgical resection. The only risk is recurrence (20 p. 100). PMID- 9747322 TI - [Diagnostic case. Primary HIV infection]. PMID- 9747323 TI - [Case for diagnosis. Herpes rugbiorum]. PMID- 9747324 TI - [Drug-induced acrosyndromes]. PMID- 9747325 TI - [Managing hyperandrogenism]. PMID- 9747326 TI - [Lingual amyloidosis]. PMID- 9747327 TI - [Elimination protocols in atopic dermatitis]. PMID- 9747328 TI - Stability of HBV DNA in cell lines and nude mouse-passaged tissues derived from human hepatocellular carcinoma. AB - Human hepatitis B virus (HBV) infection has been closely linked to the occurrence of hepatocellular carcinoma (HCC). Hepatoma cell lines and nude mouse-passaged hepatoma tissues were used in this report to study the HBV DNA status in these cells after passage. DNA was extracted from seven hepatoma cell lines and three nude mouse passaged HCC lines. Southern blot hybridization technique was performed with either cloned HBV whole genome or subgenomic DNA fragments as probes to analyze the presence of HBV DNA. Integration of HBV DNA fragments was detected in one mouse passaged tissue, R. Hybridization with HBV subgenomic DNA revealed that there were some DNA rearrangements of the integrated HBV DNA in R. However, the integrated HBV DNA could not be detected in the cell line derived from R after in vitro cultivation for 2 years. Both episomal form and integrated HBV DNA were detected in a cell line NTU-h3. Episomal form HBV DNA ih NTU-h3 changed after several passages. HBV DNA in NTU-h3 was unstable after in vitro cultivation. Therefore, we concluded that the presence of HBV DNA might not be essential for the maintenance of the tumorigenicity of hepatoma and the nude mouse system was more stable for maintaining HBV DNA in HCC. PMID- 9747329 TI - [Analysis of Salmonella serovars in Taiwan by the phase induction method]. PMID- 9747331 TI - A study on the in vitro susceptibility of Trichomonas vaginalis to metronidazole. AB - The susceptibility of 32 clinical isolates of Trichomonas vaginalis to metronidazole has been studied under aerobic and anaerobic conditions. Of 32 patients with vaginal trichomoniasis, 27 (84%) were cured by a standard metronidazole treatment regimen (200 mg thrice daily for seven days). The geometric means of minimum inhibitory concentration (MIC) under aerobic and anaerobic conditions for these isolates were 2.0 and 0.4 micrograms/ml, respectively; the geometric means of minimum lethal concentration (MLC) under aerobic and anaerobic conditions were 7.4 and 2.0 micrograms/ml, respectively. However, for those five patients with treatment failure, the geometric means of MIC for these isolates under aerobic and anaerobic conditions were 6.8 and 1.1 micrograms/ml, respectively; the geometric means of MLC under aerobic and anaerobic conditions were 18 and 5.5 micrograms/ml, respectively. Trichomonads reisolated from patients after treatment failure had similar susceptibility to metronidazole as before treatment. However, all five women were cured by a second course of metronidazole treatment. Although primary treatment failure was common when isolates of T. vaginalis had aerobic MLC values of > 18 micrograms/ml or anaerobic MLC values > 5.5 micrograms/ml, two cases with isolates having high MLC values (aerobic: 20 micrograms/ml, anaerobic: 5 micrograms/ml) responded well to the standard treatment. It was evident that no metronidazole-resistant trichomonads were found in this study. PMID- 9747330 TI - Biological and immunological studies on a low virulence isolate of the Tulahuen strain of Trypanosoma cruzi. AB - Reduced virulence for mice was characterized in an isolate (LV1) of a clone of the Tulahuen strain of Trypanosoma cruzi. LV1 caused long term chronic parasitemias which were measured for 140 days in both C3H/He and BALB/c mice inoculated with 1 x 10(5) trypanosomes/mouse. In contrast to the acute and rapidly lethal Tulahuen strain infections in both strains of mice, all of the animals survived the LV1 infections. Sera of C3H/He mice infected with the Tulahuen strain or LV1 isolate displayed similar titers in an enzyme-linked immunosorbent assay (ELISA) when reacted against homologous or heterologous extracts of epimastigote stages of the trypanosomes. Western blot reactions of the Tulahuen, Raccoon V, LV1 isolates, and the closely related European bat parasite, Trypanosoma dionisii defined shared antigens between the strains and species, while some appeared to be strain- and species-specific. The studies indicate a mutational event(s) resulted in reduced virulence and suggest that survival of the mice infected with T. cruzi is not correlated with high ELISA antibody titers. Since lower antibody titers are exhibited by mice infected with LV1 than mice infected with the Tulahuen strain, survival may be dependent on the specificities of the antibodies synthesized during the infections, cell mediated immune responses, and/or biochemical factors of the LV1 isolate which control virulence and differ from those of the original Tulahuen strain. PMID- 9747332 TI - [A rapid and simple method for direct genotype determination of ABO blood group]. AB - We report the use of an allele-specific polymerase chain reaction (ASPCR) format together with low melting temperature agarose gel electrophoresis which allows rapid identification of the six major genotypes of the ABO blood group. Four sequence specific primer sets, each specific for a different set of ABO alleles, were used. Twenty individuals, whose ABO genotypes were previously determined by serological and family analysis, were typed with this new approach. A 100% correlation between serology and the ASPCR was found. This method is rapid, simple, and reproducible. Potential applications include identification of ABO subgroups and variants, paternity testing, as well as forensic science. PMID- 9747333 TI - Bilophila wadsworthia isolated from clinical specimens in Taiwan. AB - Bilophila wadsworthia, a newly discovered anaerobic gram-negative species, has been found in such various clinical specimens as bile, pus, blood et cetera, in addition to appendicitis specimens. The formation of black center colonies in bacteroides bile esculin (BBE) agar is one of the important characteristics of the species. PMID- 9747334 TI - Identification and analyses of periodontal pathogens in Taiwan by microbiological tests. AB - The purpose of this study was to use microbiological tests for diagnosis of periodontal diseases in Taiwan. Anaerobic culture, direct microscopy, indirect immunofluorescence (IF), and biochemical tests were used to examine 336 samples for the specific microorganisms in subgingival plaque. The results indicated that gram-negative species and motile bacteria were less frequently detected, and in lower proportion, in samples from healthy sites. The bacteria found frequently in healthy group were the coccal forms. However, Bacteroides forsythus detected by IF showed a close association with periodontal inflammation. Porphyromonas gingivalis was found with about 53% frequency in the periodontitis group; in more than half the samples the proportion was above 5%. Actinobacillus actinomycetemcomitans was recovered with 48% frequency of periodontitis group. Other cultivable species including Campylobacter rectus, Capnocytophaga species, Centipeda periodontii, Eikenella corrodens, Fusobacterium nucleatum, Prevotella intermedia, Selenomonas species, and the Spirochetes were detected in a significantly higher proportion in periodontitis group. The results strongly support the use of microbiological tests as adjuncts to diagnosis, and for assessment of the importance of microbiota in periodontal disease. PMID- 9747335 TI - Susceptibility of avian mycoplasmas isolated in Taiwan to 21 antimicrobial agents. AB - Twenty-one antimicrobial agents were incorporated individually into Frey's agar to evaluate their inhibitory activities against 86 isolates of avian mycoplasmas recently detected in Taiwan. Among them, 45 and 37 isolates were found positive with Mycoplasma gallisepticum and Mycoplasma synoviae fluorescent antibody conjugate, respectively. Twenty-one other isolates were unable to be identified by the above 2 conjugates. All of the field isolates were highly sensitive (with MIC50 < 1 microgram/ml) to enrofloxacin, gentamicin, myplabin, tiamutin and tylosin. However, those field isolates were highly resistant (with MIC50 > 32 micrograms/ml) to apramycin, chlortetracycline (CTC), erythromycin (ER), flumequine (FI), nalidixic acid (NA), oxolinic acid (OA), oxytetracycline (OTC) and spiramycin (SP). The inhibitory activities of the antibiotics which possessed an MIC90 of 50 micrograms/ml or less against local isolates were, in decreasing order, enrofloxacin (< 0.004 microgram/ml), gentamicin (1.53 micrograms/ml), tiamutin (1.81 micrograms/ml), tylosin (3.2 micrograms/ml), streptomycin (SM; 12.0 micrograms/ml), colistin (13.1 micrograms/ml), chloramphenicol (14.0 micrograms/ml), spectinomycin (15.0 micrograms/ml), myplabin (16.0 micrograms/ml), spiramycin (30.0 micrograms/ml), minocycline (32.0 micrograms/ml). The MIC90 of OA, CTC, SM, FI, SP, OTC, ER or NA was greater than 50 micrograms/ml; which work poorly in the control of mycoplasmoses. Since the antibiotic control policy is quite loose in Taiwan, many antimicrobial agents are often freely used in clinics, with a resulting gradual decrease in the inhibitory activity to the avian mycoplasmas. PMID- 9747336 TI - Partial nucleotide sequence of Japanese encephalitis virus ling strain genome and comparison of the encoded structural proteins and nonstructural protein NS1 among Japanese encephalitis virus strains. AB - Approximately 4000 nucleotides of the 5'-terminal portion of Japanese encephalitis virus (JEV) Ling strain genome were cloned and sequenced. This nucleotide sequence and its encoded C-PrM-E-NS1 polyprotein sequences were also compared with the corresponding sequences of other JE virus strains. Results demonstrated that the nucleotide sequence homology varied from 97.1 to 99.3% and the amino acid homology 98.6 to 98.9%. Based on homology, the Ling strain was closer to the Beijing-1 strain than to the SA14 and JaOArS982 strains. However, only on comparison of the E sequence, which neutralization, hemagglutination inhibition and complement fixation antigenic determinants are located, between Ling and other JEV strains demonstrated that nucleotide sequence homology varied from 97.1% to 99.3% and amino acid homology from 98.6% to 99.2%. The Ling strain JEV is more closely related to the Beijing-1 strain than to the Nakayama NIH, SA14 and JaOArS982 strains in that order. Based on this analysis, the Taiwanese JEV strain appears to be more closely related to the Chinese strain than to the Japanese strain. Also, JEV strains isolated in humans are more closely related to each other than to JEV strains of mosquito isolates. PMID- 9747337 TI - [Preliminary surveillance of tetanus antitoxin in Taiwan]. PMID- 9747339 TI - [Seroprevalence of human immunodeficiency virus infection in Guinea-Bissau, west Africa]. AB - The seroprevalence of human immunodeficiency virus (HIV) infection in Guinea Bissau, West Africa, was determined by enzyme-linked immunosorbent assay (ELISA). From January 1987 to February 1993, 590 patients from the outpatient and inpatient departments of Regional Hospital at Canchungo, Cacheu, Guinea-Bissau were studied. The overall seropositive rate was 16%. Patients in the age between 25 and 54 accounted for 78% of HIV-positive cases. The seropositive rate according to the diagnosis was: 6% in pregnant women, 40% in patients with gonorrhea/syphilis, 14% in patients with vaginitis and 22% in patients with active pulmonary tuberculosis. The seropositivity for HIV-1/2 in the pregnant women might reflect the seroprevalence in general population of Guinea-Bissau. Accordingly, the estimated population infected by HIV would be sixty thousands in Guinea-Bissau. Both sexually-transmitted diseases and tuberculosis were the risk factors for HIV infection. This study shows that HIV infection is a critical problem of public health in Guinea-Bissau. Strategies to prevent the seeding of HIV are of great importance. Moreover, the members of medical mission from our country must keep alert for preventing HIV infection. PMID- 9747338 TI - [A simplified emulsification method for antigens with oil adjuvants]. AB - In order to enhance the immune response, people usually add adjuvant to the antigen in immunization. Freund's adjuvant is one of the most used. It can prolong the duration of the antigen staying in the body of the animal, and through continuous stimulation of the antigen the production of antibody is increased. This paper describes a few points in facilitating the emulsification of the antigen and making it more effective. The process can be summarized as follows: (1) Force a small amount of adjuvant from syringe B by pushing it into syringe A filled with antigen solution to mingle with the latter. (2) Push the same volume of the mix from syringe A back to syringe B slowly. (3) Repeat the above mixing process until the mixed portion has become milky white. (4) Gradually increase the volume by small amounts and each time do the same mixing until final completion. After immunization with the mixture, potent sera were always obtained. It has become a routine in this laboratory to use such mixture in its immunization tasks. PMID- 9747340 TI - A rapid method for determination of susceptibility of Mycobacterium tuberculosis to isoniazid, using acridinium-ester-labeled DNA probe. AB - A rapid isoniazid (INH) susceptibility test was developed for Mycobacterium tuberculosis by using acridinium-ester (AE)-labeled DNA probe. The method was applied to two reference strains and 20 clinical isolates, then compared with the standard proportion method. By comparing the difference of log relative light units (RLUs) M. tuberculosis cultures incubating in the presence and absence of INH, INH susceptibility could be determined with the AE-DNA probe after three to five days of incubation. The difference of log RLUs of susceptible strains was statistically significantly lower than that for resistant strains after incubation for three to five days. The cutoff value was defined as "mean + one standard deviation" of the difference between log RLU(INH) and log RLU(no INH) on Day 5. By this criterion, agreement between the AE-DNA probe and the proportion concentration method was found in 19 of 20 susceptibility tests (95%). The AE-DNA probe test is rapid and non-isotopic, and may provide a useful alternative method for INH susceptibility testing for clinical isolates of M. tuberculosis. PMID- 9747341 TI - Partial purification of hydrogenase from Anabaena sp. CH3 with heat treatment. AB - The methylviologen-dependent hydrogenase of Anabaena sp. CH3 is unstable at 4 degrees C and in air. All the purifying procedures were carried out at 25 degrees C and under argon atmosphere. The enzyme was partially purified by the following steps: heat treatment, DEAE-cellulose ion-exchange chromatography and gel filtration on TSK-Fractogel. Experimental results indicated that the heat treatment procedure was beneficial for the separation of the enzyme from phycobiliproteins. PMID- 9747342 TI - Lipopolysaccharide binding and antibacterial activities of a synthetic peptide representing amino acids 90-101 of bactericidal/permeability-increasing protein. AB - The bactericidal/permeability-increasing protein (BPI) of polymorphonuclear leukocytes is a potent antibacterial agent specific for gram-negative bacteria. BPI can bind to lipopolysaccharide (LPS) and neutralize its toxicity. However, little is known about the specific site and mechanisms of the BPI involved in this LPS binding and antibacterial activities. This study compared the amino acid sequences among BPI, cecropin A, magainin 2, and polymyxin B, and identified a common structure among these four bactericidal agents. They share a basic amphipathic alpha helix motif (Baah). A short peptide that represents amino acids 90-101 of BPI was then synthesized to test if it possessed any LPS binding and antibacterial activities. Results from in vitro lymphocyte culture indicated this peptide was able to inhibit LPS-induced lymphocyte proliferation, suggesting that it may interact with LPS. This LPS binding ability of BPI peptide 90-101 was further supported by the results from HPLC assays which showed the mobility of the peptide shifted in the presence of LPS. Furthermore, the antibacterial spectra of this peptide and cecropin peptide 1-11 were very similar to that of polymyxin B, even though the antibacterial activities of these two peptides were less potent than that of polymyxin B. In addition, the antibacterial activities of these two peptides and polymyxin B were inhibited by free LPS or a high concentration of MgCl2. These results thus suggest that a common structure (Baah) and antibacterial mechanism may be involved in these antibacterial agents. PMID- 9747343 TI - [Assessment of a new four-hour diagnostic kit--RapID onE system for the identification of enteric bacteria]. AB - RapID onE System is a newly developed four-hour rapid diagnostic kit for the identification of enteric bacteria. To know the effectiveness of this system, we used 125 strains of oxidase-negative, gram-negative bacilli for this evaluation. Except for Acinetobacter calcoaceticus, all the bacilli belong to family Enterobacteriaceae. The bacterial strains of this assessment belong to 12 genera and 20 species. Among them, 84 strains were freshly isolated from clinical specimens and 41 strains were frozen (-70 degrees C) stock clinical isolates. The results show that 115 (92.0%) strains were correctly identifed to the species level. It yielded 92.9% and 90.2% of correct identification of fresh isolates and frozen stocks, respectively. In this paper, the reading criteria of RapID onE System would also be discussed. PMID- 9747344 TI - Allopurinol hypersensitivity syndrome. AB - Allopurinol hypersensitivity syndrome (AHS) is an infrequent but life-threatening adverse reaction of allopurinol therapy. The records of 38 patients with the allopurinol hypersensitivity syndrome evaluated at the Veterans General Hospital Taipei were reviewed. The clinical pictures included fever, rash, leukocytosis, eosinophilia, impaired renal function and hepatocellular injury. Nine patients died (24%) and the major cause of death was infection. The use of corticosteroids increased neither survival nor mortality rate. Twenty-six percent of patients were treated with allopurinol for asymptomatic hyperuricemia, which was not an established indication of the drug, should be avoided. The most important factor of mortality was toxic epidermal necrolysis (TEN) (p < 0.001 compared with other skin lesions). As there is no way to identify the risk group of patients or to make effective treatment for AHS, the only means of minimizing the incidence of AHS is to limit the allopurinol therapy to accepted indications and to adjust the dosage for the patient's renal function. PMID- 9747345 TI - Isolation of Bacillus cereus in the feces of healthy adults in Taipei City. AB - Fecal specimens from 100 healthy adults were collected and examined for the presence of Bacillus cereus which has been associated with 28% of the outbreaks of food poisoning on Taiwan within the last 3 years. Total isolate rate from these specimens was 8%. Variations in isolation rates were found not only in sexes, but also in different age-groups. Therefore, presence of B. cereus in the feces of healthy adults may be unpredictable and relate to foods consumed or to other factors. Obviously, an isolation rate of B. cereus as high as 30% during the outbreak investigation is still not a strong evidence to implicate this organism as an etiological agent. PMID- 9747346 TI - Development of new functional immobilized microbial cell systems and their applications. AB - Immobilized cell technology has been playing a vital role in the development of fermentation processes. For the past several years, I have been working on immobilized cell systems with an aim of developing novel immobilized biosystems where physical, chemical as well as biological functions are incorporated into the immobilization carrier. By efficiently integrating these new functions with the innate abilities of immobilized cells, the area where immobilized cell systems can be utilized will expand, and the process efficiency will be greatly improved. PMID- 9747347 TI - Identification and characterization of a protease produced by Vibrio parahaemolyticus in iron-limited medium. AB - Two proteolytic proteins (about 43 and 90 kDa) were produced by clinical strains of Vibrio parahaemolyticus cultured in iron-limited medium. The 43 kDa-protease was partially purified by ammonium sulfate precipitation, ultrafiltration fractionation and DEAE-Sephacel chromatography. This protease had an optimum pH range of 7 to 8, and an optimum reaction temperature of about 40 degrees C. It was heat-labile, being partially inactivated by heat-treatment at 60 or 90 degrees C for 10 min. The protease hydrolyzed casein, gelatin, elastin, collagen and hemoglobin. As a chymotrypsin-like protease, it was inhibited only by the chymostatin among seven protease inhibitors tested. Activity of this protease was partially inhibited by 1 mM of Co2+, Cu2+, Zn2+ and Hg2+ and slightly enhanced by Ca2+ and Ba2+. It was completely inactivated by orthophenanthroline (OPA), and the OPA-inactivated sample was partially reactivated by Ca2+ and Fe2+. In conclusion, this 43-kDa protease of V. parahaemolyticus was an unstable neutral chymotrypsin-like metalloprotease; Ca2+ and/or Fe2+ was essential for its activity or stability. PMID- 9747348 TI - Clindamycin resistance transfer in Bacteroides fragilis. AB - Clindamycin is one of the antimicrobial agents most commonly used against anaerobes. Resistance to clindamycin in Bacteroides fragilis has been increasing recently. Thirty strains of clindamycin-resistant (including multi-resistant) B. fragilis were collected for study of cross-resistance to beta-lactam agents and beta-lactam--beta-lactamase inhibitor and resistance transferability. Imipenem was the most active drug against these 30 isolates. Resistance to clindamycin was transferred to a recipient in 12 out of 30 donor strains by using filter-mating. Of 12 transconjugants, only three had detectable plasmids by alkaline lysis method and the remaining nine strains lacked plasmids. The transfer frequencies ranged from 10(-4) to 10(-6). The role of plasmid in the resistance transfer was not certain. However, the results suggest that non-plasmid-mediated transfer accounted for the majority of the transfers of clindamycin-resistance of B. fragilis in this study. Tetracycline resistance was co-transferred from six donors. There was no evidence of co-transference of beta-lactam resistance under the selection marker of clindamycin, beta-lactam, or both. Therefore, non-plasmid mediated transfer may play an important role in dissemination of resistance transfer in B. fragilis in Taiwan. PMID- 9747350 TI - Light-emitting aeromonas and plesiomonas generated by transconjugating luxAB from Escherichia coli. AB - The transposon derivative has been placed on a transposition suicide vector to yield pDB30 in Escherichia coli WA803. A simple method, using a Tn5 derivative Tn5-Lux, has been successfully devised for the introduction and stable expression of the bioluminescence property in Pseudomonas sp., Agrobacterium sp., and Rhizobium sp. In this study, there was also successful mating between Escherichia coli WA803(pDB30) and strains of Acromonas hydrophila and Plesiomonas shigelloides. These bacteria emitted bioluminescence after they gained pDB30 by transconjugation. PMID- 9747349 TI - [Screening and rapid identification of Bacillus thuringiensis mutants]. AB - Mutants of Bacillus thuringiensis subsp. kurstaki NTU 9 and Bt 158, which were isolated previously for using the diamondback moth as a target insect in Taiwan, were screening by either protein electrophoresis of intracellular proteins or enzyme-linked immunosorbent assay (ELISA). The optimal conditions of effective protein electrophoresis were (1) 24-hour cells harvested from nutrient broth were crashed by petite glass beads followed by centrifugation. And (2) the supernatant pretreated by heating at 60 degrees C for 2 minutes was electrophoresed with 7.5% native PAGE at 110 voltages. On ELISA, the antiserum used was obtained from rabbits immunized with Bt 158 crystal protein. Optimal antigen coating concentration of ELISA, attained by chequer-board titration method, was 10 micrograms/ml. Antigens (crystal protein) in samples were detected by competitive inhibition method with antiserum diluted to 10(4) fold. By using protein electrophoresis and ELISA methods, two isolates A 71 and BN 11, were denoted respectively as qualitative and quantitative mutants of Bacillus thuringiensis. PMID- 9747351 TI - A preliminary report on Borrelia burgdorferi infection in the Taiwan area. AB - A total of 273 serum specimens from different areas and sources were tested against Borrelia burgdorferi antigens by enzyme-linked immunosorbent assay method. Positive rates of serological reactions were 3% and 58% for healthy persons and syphilis patients, respectively. Obviously, there was a lot of cross reaction in the venereal disease group. Meanwhile, positive rates were 3% and 13% in the sera collected from Taiwan and Orchid Island, respectively. This difference may reflect a less developed environment in the latter. Since reported cases of Lyme disease in Taiwan are rare, serologic tests are usually adapted for rapid diagnosis in common laboratories. As for disease confirmation, clinical observations, epidemiological data and exposure in an endemic area must also be considered. PMID- 9747353 TI - Distinct patterns of collagen gene expression are seen in normal and keloid fibroblasts grown in three-dimensional culture. AB - The purpose of this study was to examine collagen gene expression in various types of scar fibroblasts as well as normal fibroblasts in a novel three dimensional culture system and to compare them with those in a monolayer culture system. Cells in three-dimensional culture formed multiple layers within the self produced dense extracellular matrix and formed a dermis-like structure. In monolayer culture, both normal and scar fibroblasts continued to express high levels of mRNA for pro alpha 1(I) and pro alpha 1(III) collagens. However, in three-dimensional culture, the mRNA levels gradually declined in normal fibroblasts. In contrast, mRNA levels remained high in keloid and hypertrophic scar fibroblasts. Atrophic scar fibroblasts demonstrated similar changes to normal fibroblasts in three-dimensional culture. When we compared mRNA expression in fibroblasts from the centre and the edge of hypertrophic scar, cells from the centre showed a persistently decreased level of collagenase mRNA expression. These results suggest that the mRNA expression pattern of pro alpha 1(I) and pro alpha 1(III) collagens varies depending on the culture system. Fibroblasts from keloids and hypertrophic scar may have a defective system of down-regulation in extracellular matrix metabolism. PMID- 9747352 TI - Fibrillin microfibrils are reduced in skin exhibiting striae distensae. AB - Striae distensae (striae: stretch marks) are a common disfiguring condition associated with continuous and progressive stretching of the skin--as occurs during pregnancy. The pathogenesis of striae is unknown but probably relates to changes in those structures that provide skin with its tensile strength and elasticity. Such structures are components of the extracellular matrix, including fibrillin, elastin and collagens. Using a variety of histological techniques, we assessed the distribution of these extracellular matrix components in skin affected by striae. Pregnant women were assessed for the presence of striae, and punch biopsies were obtained from lesional striae and adjacent normal skin. Biopsies were processed for electron microscopy, light microscopy and immunohistochemistry. For histological examination, 7 microns frozen sections were stained so as to identify the elastic fibre network and glycosaminoglycans. Biopsies were also examined with a panel of polyclonal antibodies against collagens I and III, and fibrillin and elastin. Ultrastructural analysis revealed alterations in the appearance of skin affected by striae compared with that of normal skin in that the dermal matrix of striae was looser and more floccular. Light microscopy revealed an increase in glycosaminoglycan content in striae. Furthermore, the number of vertical fibrillin fibres subjacent to the dermal epidermal junction (DEJ) and elastin fibres in the papillary dermis was significantly reduced in striae compared with normal skin. The orientation of elastin and fibrillin fibres in the deep dermis showed realignment in that the fibres ran parallel to the DEJ. However, no significant alterations were observed in any other extracellular matrix components. This study identifies a reorganization and diminution of the elastic fibre network of skin affected by striae. Continuous strain on the dermal extracellular matrix, as occurs during pregnancy, may remodel the elastic fibre network in susceptible individuals and manifest clinically as striae distensae. PMID- 9747354 TI - Human keratinocytes express transcripts for three isoforms of parathyroid hormone related protein (PTHrP), but not for the parathyroid hormone/PTHrP receptor: effects of 1,25(OH)2 vitamin D3. AB - Parathyroid hormone-related protein (PTHrP) is strongly expressed in the epidermis and has been implicated in the regulation of growth and differentiation of keratinocytes. PTHrP has N-terminal sequence homology with parathyroid hormone (PTH) and binds to the type I PTH/PTHrP receptor, but earlier reports suggest that keratinocytes do not possess this cell surface receptor. In order to determine which PTHrP mRNA isoforms are expressed by keratinocytes and whether the type I receptor mRNA is present, we designed specific primers for reverse transcriptase-polymerase chain reaction. The interaction of PTHrP with other promoters of keratinocyte differentiation is unclear. In particular, 1,25(OH)2D3 is also fundamental in calcium homeostasis and induces changes in intracellular calcium. We therefore investigated the effect of 1,25(OH)2D3 on PTHrP mRNA expression and protein production in cultured human keratinocytes. Cells were incubated for 3 days at concentrations of 1.25(OH)2D3 of 10(-10)-10(-6) mol/L. PTHrP in culture supernatant, measured by two site immunoradiometric assay, was 915 +/- 98 PTHrP fmol/mg of cell layer protein in untreated cultures decreasing to 570 +/- 113 with 10(-8) mol/L and 402 +/- 24 with 10(-6) mol/L 1,25(OH)2D3 (mean +/- SEM, P < 0.01, n = 6). Transcripts for all three PTHrP isoforms (139, 141 and 173 amino acids) were detectable in keratinocyte mRNA. Corresponding to the decrease in PTHrP protein we demonstrated a reduction in all three PTHrP mRNA transcripts after 3 days' incubation with 1,25(OH)2D3 over a concentration range 10(-10)-10(-6) mol/L. Repeated studies failed to detect type I PTH/PTHrP receptor mRNA in human keratinocytes, either in control cultures or in the presence of 1,25(OH)2D3. We have shown that keratinocytes produce abundant PTHrP and that this is modulated by 1,25(OH)2D3, suggesting a physiological role. Further studies are required to investigate the relative expression of PTHrP isoforms, their role in keratinocyte signalling and the receptors involved. PMID- 9747355 TI - Erythema multiforme lesions are associated with expression of a herpes simplex virus (HSV) gene and qualitative alterations in the HSV-specific T-cell response. AB - A common form of erythema multiforme, herpes-associated erythema multiforme (HAEM), occurs following infection with herpes simplex virus (HSV). Here we report that HSV gene expression and the qualitative nature of the virus-specific T-cell responses are related to HAEM lesion development. Skin from HAEM lesions and 1-3 months healed HAEM lesional skin were positive for the viral DNA polymerase gene (Pol) by polymerase chain reaction. However, gene expression as determined by immunohistochemistry with Pol-specific antibody was seen only in HAEM lesions, suggesting that lesion development is associated with Pol gene expression. Similar HSV-specific T-cell lymphoproliferative responses were seen in peripheral blood mononuclear cells (PBMCs) from patients with acute or healed HAEM lesions or HSV lesions and from HSV-seropositive patients with unrelated inflammatory diseases. However, the T-cell receptor variable (V beta) chain repertoire of HSV-stimulated PBMCs obtained from HAEM lesions was altered; the prevalence of some families of variable chain (namely V beta 16 and V beta 19) was reduced, whereas the prevalence of others was increased (namely V beta 2 and V beta 7). V beta 2 cells were found in HAEM lesional skin positive for Pol antigen, suggesting that these cells home to viral antigen-positive skin. PMID- 9747356 TI - Precise ultrastructural localization of in vivo deposited IgG antibodies in fresh perilesional skin of patients with bullous pemphigoid. AB - Bullous pemphigoid (BP) is a blistering skin disease in which the patient develops autoantibodies to the epidermal basement membrane zone. Using postembedding immunogold electron microscopy, we previously demonstrated that autoantibodies against the 230-kDa BP antigen (BPAG1) bind to the intracellular hemidesmosomal component of normal skin, whereas those against the 180-kDa BP antigen (BPAG2) bind only along the plasma membrane of hemidesmosomes. The purpose of the present study was to elucidate the precise localization of the in vivo deposited IgG antibodies in fresh perilesional skin of patients with BP. Samples of fresh perilesional skin were obtained from three patients with BP whose sera reacted only with BPAG1, only with BPAG2, and with both BPAG1 and BPAG2 upon immunoblotting using epidermal extracts. Cryofixed and cryosubstituted skin samples were used as a substrate for on-surface immunolabelling. In all three cases, most of the gold particles were observed close to the plasma membrane of the basal keratinocytes. A quantitative analysis revealed that most (> 80%) of the in vivo deposited IgG antibodies in the three cases were localized within 10 nm inside to 50 nm outside of the cell membrane, with a single peak observed 0-10 nm outside of the cells (> 50%). This distribution corresponded to the location of BPAG2, but not to that of BPAG1. These findings suggest that most, if not all, of the in vivo deposited IgG antibodies in the perilesional skin of BP are directed against BPAG2, rather than against BPAG1, thus further supporting the crucial role of anti-BPAG2 autoantibody in the initial stage of blister formation in BP. PMID- 9747357 TI - Lichen planus pemphigoides is a heterogeneous disease: a report of five cases studied by immunoelectron microscopy. AB - Lichen planus pemphigoides (LPP) is a rare and controversial disease. It is characterized by bullae arising on lichen planus papules and on uninvolved skin, subepidermal bullae in histology, and linear deposits of IgG and C3 along the basal membrane zone on immunofluorescence of peribullous skin. Our goal was to identify the localization of the target antigen in cases of LPP. Five patients diagnosed with LPP on clinical, histological and immunofluorescence criteria were explored by immunoelectron microscopy and immunoblot. Our results show that the target antigen in LPP is not unique. The localization of the immune deposits was consistent with a diagnosis of bullous pemphigoid in two cases, of cicatricial pemphigoid in two cases and of epidermolysis bullosa acquisita in one case. Our study supports the view that LPP is a heterogeneous condition in which lichen planus may induce different subepidermal acquired bullous dermatoses. PMID- 9747358 TI - Immunoreactivity of alpha-melanocyte-stimulating hormone, adrenocorticotrophic hormone and beta-endorphin in cutaneous malignant melanoma and benign melanocytic naevi. AB - Melanocyte-stimulating hormone (MSH) has been reported to enhance the experimental metastatic behaviour of melanoma cells in the mouse model. alpha-MSH production and MSH receptor (melanocortin 1 receptor gene) expression have been detected in cultured normal human melanocytes and metastasized melanomas. The exact role of MSH in the metastatic behaviour of human melanoma cells is, however, not yet known. To clarify a possible role of proopiomelanocortin (POMC) derived peptides, including alpha-MSH, in melanoma development and progression, we analysed immunohistochemically the localization of alpha-MSH adrenocorticotrophic hormone (ACTH) and beta-endorphin in various kinds of benign pigmented naevocytic lesions and malignant melanomas. Three of 21 samples of common and dysplastic naevi showed detectable alpha-MSH staining in naevus cells, and five and six of 15 samples were weakly positive for ACTH and beta-endorphin staining, respectively. In melanoma samples, 24 of 45, 23 of 39 and 30 of 42 samples showed positive staining with alpha-MSH, ACTH and beta-endorphin antibodies, respectively. Furthermore, staining for all three antibodies was noted to be more intense and diffuse in samples of nodular melanoma, vertically growing acral lentiginous melanoma and superficial spreading melanoma as well as metastatic lesions compared with those of naevi. Although it is yet to be determined whether or not this strong staining for POMC-derived peptides in advanced melanoma cells indicates a role of autocrine or paracrine regulation, our results suggest a possible involvement of POMC gene products in melanoma progression. PMID- 9747360 TI - Influence of genetic and environmental factors on melanocytic naevi: a lesson from Turner's syndrome. AB - Females with Turner syndrome (TS) are alleged to have increased numbers of melanocytic naevi. Although a high count of acquired melanocytic naevi (AMN) is one of the major risk factors for melanoma, this malignancy has been reported only rarely in patients with TS. The purpose of this study was to explore the effects of environmental and genetic factors on AMN count and density in TS. AMN count and density in 24 patients with TS treated with growth hormone (GH). 24 GH treated females with GH deficiency (GHD) and 24 normal females were compared in a cross-sectional study. The average AMN density in TS was 50 naevi/m2 as compared with 18 naevi/m2 in the GHD group and 24 naevi/m2 in normal controls (P = 0.001 and P = 0.004, respectively). Duration of GH therapy did not correlate with AMN count (P = 0.44) or AMN density (P = 0.81). The pattern of distribution of naevi between constantly exposed, intermittently exposed and unexposed skin was similar in all groups. Sun exposure was the major factor that affected the regional AMN densities in the control groups, but not in the TS group. The findings of our study indicate that the effects of environmental factors on AMN count and density may vary among genetically different populations. A review of the literature suggested that melanoma is no more prevalent in TS than in the general population. PMID- 9747359 TI - Urocanic acid isomers in patients with basal cell carcinoma and cutaneous malignant melanoma. AB - Urocanic acid (UCA) is a major chromophore for ultraviolet (UV) radiation in the skin. On UV exposure, the naturally occurring trans-isomer converts to the cis isomer in a dose-dependent manner. Accumulating evidence indicates that cis-UCA acts as an initiator of the UV-induced suppression of certain skin immune functions. This immunomodulation is recognized as an important factor in the development of skin cancer. In this study, pigmentation and UCA isomers were measured in 29 patients with previous basal cell carcinoma (BCC), 23 patients with previous cutaneous malignant melanoma (MM), and 32 healthy controls. Measurements were performed on UV-exposed (forehead, upper back) and UV non exposed (buttock) skin. No significant differences in pigmentation percentage, total UCA concentration, relative (%) or absolute (nmol/cm2) cis-UCA concentration were observed between the groups in any of the body sites studied. The net production of cis-UCA after irradiation with a single test UV dose was evaluated. The relative production of cis-UCA following irradiation was significantly higher in both cancer groups when compared with the control group, while no significant difference was found between the BCC and the MM patients. PMID- 9747361 TI - Malignant melanoma in Cape Town, South Africa. AB - There is a world-wide increase in the incidence of cutaneous malignant melanoma among white people. Absence of accurate population-based data on the incidence of melanoma in South Africa prompted a study to determine the incidence, anatomical sites and pathological details of melanoma in Cape Town. In a prospective study from 1 January 1990 to 31 December 1995, all the histopathology reports of melanoma presenting in a geographically defined area of Cape Town, were actively retrieved from every pathologist practising in this area. The data evaluated included information on age, sex, ethnic group and location of residence. Details of melanoma comprised body site, Clark level of invasion, Breslow thickness in millimetres and histogenetic type. The histology slides were reviewed by a panel in those cases where the recorded information was ambiguous or incomplete. A final number of 595 reports of primary invasive cutaneous melanomas in white people was analysed. Of these 50.3% were men and 49.7% women. The overall age standardized incidence rate was 24.4 per 100,000 per annum (27.5 for men and 22.2 for women). There was no change in the incidence rate over the study period. Most melanomas in both sexes (74% of women and 71% of men) were < 1.5 mm Breslow thickness. Results of this study indicate a high incidence rate of melanoma in white South Africans, comparable with that in Australia, which demands urgent preventive health measures. PMID- 9747362 TI - Mohs' surgery of periocular basal cell carcinoma using formalin-fixed sections and delayed closure. AB - Mohs' surgery of periocular basal cell carcinoma (BCC) ensures a high cure rate with maximal preservation of normal tissue. The formalin-fixed paraffin-embedded tissue technique allows Mohs' surgery to be performed using routine pathology facilities and permits the efficient use of operating room personnel and theatre time. The inevitable delay between excision and closure may potentially result in a poor functional and cosmetic outcome, particularly around the eye. We prospectively studied all patients with periocular BCC treated with this technique at our unit between 1985 and 1996. One hundred and twenty-three periocular BCCs in 120 patients were treated. Microscopic clearance was achieved in all cases. Closure was performed on average 5 days after the initial excisional stage. Closing procedures included direct closure, flaps and grafts. Significant complications affecting outcome were noted in only two patients. Eighty-eight per cent of patients assessed had a functional and cosmetic result regarded as excellent, good or adequate. Mohs' surgery of periocular BCC using formalin-fixed paraffin-embedded tissue and delayed closure results in a satisfactory functional and cosmetic outcome and offers a viable alternative to the frozen section fresh tissue technique. PMID- 9747363 TI - Ruby laser treatment for hirsutism: clinical response and patient tolerance. AB - We report the results of a study of the ruby laser in normal mode for treatment of hirsutism in 43 patients with skin types I-IV. A pulse width of 950 microseconds, a 4 mm spot and a fluence sufficient to produce minimal whitening of the epidermis were used (mean 48 J/cm2). In each patient, one site was treated once only, and a second site received four treatments at monthly intervals. After a single treatment, hair counts had reduced to a mean of 46% (median 67, reduced to 26) of the pretreatment values at 1 month, but increased to 80% (median 42) at 3 months and were 66% (median 37) at 6 months. One month after three treatments, hair counts had reduced to a mean of 29% (median 60, reduced to 15) of pretreatment values, and 3 months after four treatments at the same site, counts had increased to 44% (median 21). Patient tolerance of the treatment was good and higher fluences tended to be tolerated at consecutive visits. The first treatment was followed by mild, short-lived erythema and swelling in 60% (25 of 42) patients. Fourteen per cent (six of 42) experienced mild blistering and 33% (14 of 42) crusting. More severe reactions were seen infrequently. These results indicate that normal mode ruby laser treatment is well tolerated. Sustained reduction in hair counts can be achieved for at least 6 months, and multiple treatments produce greater clinical effects. PMID- 9747364 TI - Porphyria cutanea tarda and hepatitis C virus infection. AB - We studied the prevalence of hepatitis C virus (HCV) infection in 20 Japanese patients with sporadic-type porphyria cutanea tarda (PCT). Seventeen of the 20 patients (85%) had anti-HCV antibodies. Biochemical remission was observed in nine patients, six of whom still had positive HCV RNA copies. These results suggest that HCV infection is a triggering factor for PCT in Japan. However, continuous HCV infection seems to exert little influence on the maintenance of abnormal porphyrin metabolism. Hepatocellular carcinoma (HCC) developed in five of the 17 HCV-positive patients, three of whose PCT was in remission. Four of these patients showed chronic active hepatitis or cirrhosis on liver biopsy. PCT patients with HCV infection should be followed up long-term because of the possibility of HCC. To evaluate the risk of HCC, liver biopsy may be required, even when the patient is in biochemical remission. PMID- 9747365 TI - Patch testing for Compositae allergy. AB - The main allergenic constituents of Compositae plants are the sesquiterpene lactones (SLs). In recent years, a mixture of three SLs, each with a different sesquiterpene skeleton (alantolactone, dehydrocostuslactone or costunolide), has been routinely used to detect Compositae allergy. The purpose of our study was to establish the value of testing with a range of Compositae extracts. Ninety-seven consecutive patients with exposure pattern dermatitis or hand eczema and gardening as a hobby or occupation were patch tested to the European standard series including SL mix 0.1% pet., Compositae extracts and other relevant allergens. Twenty-six of the 97 patients tested showed allergic reactions to SL mix or Compositae extracts. Fifteen of these showed positive reactions to both the SL mix and Compositae extracts. Eleven patients showed a negative reaction to the SL mix but positive reaction to the Compositae extracts. Dandelion elicited a positive reaction in eight of the 11 SL mix-negative patients and three patients reacted to feverfew. The SL mix failed to detect 38% of our Compositae-sensitive patients. Dandelion extract alone detected 73% of SL-negative patients. Additional testing with feverfew extract would have detected 91% of the SL mix negative individuals. Our study highlights the importance of testing the response of SL-negative patients to additional Compositae extracts when there is a clinical suspicion of Compositae allergy. PMID- 9747366 TI - Flucloxacillin in the treatment of atopic dermatitis. AB - Although colonization of atopic dermatitis by Staphylococcus aureus is universal and bacterial infection is common, it is not known whether antibiotic therapy is helpful in eczematous children who do not have any signs suggestive of bacterial infection. Fifty children aged 1-16 years with atopic dermatitis took part in a randomized double-blind placebo-controlled study of 4 weeks treatment with oral flucloxacillin, with an 8-week follow-up period. The change in the mean of the log10 of the counts/cm2 of S. aureus after 4 weeks of treatment was significantly different for patients receiving treatment, compared with the change for those receiving the placebo (P = 0.008). However, the difference in the change at 14 days after stopping treatment was not significant (P = 0.32). Methicillin resistant strains of S. aureus were cultured from five children during or after treatment. Flucloxacillin did not improve the symptoms or clinical appearance of atopic dermatitis and only temporarily changed skin colonization by S. aureus. PMID- 9747367 TI - Delusions of parasitosis. A psychiatric disorder to be treated by dermatologists? An analysis of 33 patients. AB - Delusions of parasitosis is a rare disorder in which patients have the false and fixed belief that they are infested by parasites. It is a psychiatric disorder, but patients usually present to a dermatologist and referral to a psychiatrist is almost always rejected. Treating a patient with delusions of parasitosis requires patience and tact. The neuroleptic pimozide is the treatment of choice, but a significant problem is convincing the patient to take the drug. We report a study of 33 patients (13 men and 20 women) with delusions of parasitosis. The mean age at onset was 56.9 years and the mean duration of symptoms before attending the department of dermatology was 1.3 years. Pimozide (Orap) was prescribed for 24 patients, but only 18 patients took it. Follow-up information was available for 18 patients: five had full remission, four were less symptomatic, five were unchanged and four had died of unrelated causes. PMID- 9747368 TI - 8-hydroxydeoxyguanosine in urine as an index of oxidative damage to DNA in the evaluation of atopic dermatitis. AB - 8-hydroxydeoxyguanosine (8-OHdG) is one of the products which are excreted in urine as a result of oxidative damage to DNA. We investigated the feasibility of using 8-OHdG in urine as an index for oxidative damage to DNA in atopic dermatitis (AD). Seventeen patients with long-standing AD and 17 healthy volunteers were enrolled in this study. The severity of AD was evaluated by SCORAD index. Eosinophils, total IgE and lactate dehydrogenase-5 in peripheral blood were measured as clinical parameters for AD. A newly developed enzyme linked immunosorbent assay method was used to measure urine 8-OHdG. The AD patients showed significantly higher levels (P < 0.0001) of 8-OHdG in their urine than corresponding controls. Urine 8-OHdG levels showed as strong a positive correlation as other haematological parameters did using the SCORAD index. Thus, we conclude that the urine 8-OHdG levels can also serve as a biochemical index of tissue damage and can act as a useful tool in the clinical evaluation of AD. PMID- 9747369 TI - Phagocytosis and oxidative burst by neutrophils in patients with recurrent furunculosis. AB - Neutrophil phagocytosis of fluorescently labelled Staphylococcus aureus and oxidative burst by the neutrophils were assessed by flow cytometry in 22 patients with recurrent furunculosis and in 17 controls. Phagocytosis and oxidative burst were not found to be significantly different between the patients and controls. Low serum iron concentrations were demonstrated in six patients (27%). In these patients with hypoferraemia, oxidative burst was significantly lower than in the patients without hypoferraemia and in the controls. These data suggest that hypoferraemia may be an important predisposing factor in a subgroup of patients with recurrent furunculosis in impairing oxidative killing by neutrophils. PMID- 9747370 TI - The significance of the 'dust-like particles' pattern of immunofluorescence. A study of 66 cases. AB - Subacute cutaneous lupus erythematosus (SCLE) is a marker of a unique subset of lupus erythematosus patients. A 'dust-like particles' direct immunofluorescence (DIF) staining pattern, which consists of fine granular particles of immunoglobulin(s) scattered through the epidermis and the cellular infiltrates of the dermis, was reported to be specific for SCLE. In this study, we assessed the real specificity of this staining pattern, which had not yet been evaluated. We systematically searched for the dust-like particles staining pattern among the 4374 skin biopsy specimens submitted for direct cutaneous immunofluorescence during a 7-year period (1989-96). The corresponding patient records were reviewed. Dust-like particles were observed in 66 samples originating from 60 patients. Only 53% of the patients had SCLE. The remaining patients had systemic lupus erythematosus with visceral involvement (17%), discoid lupus erythematosus (3%), mixed connective tissue disease (2%). Sjogren syndrome (2%) and other diseases. Eighty-five per cent of the patients had connective tissue disease. Seventy-seven per cent of the patients were positive for antinuclear antibodies, but only 36% were positive for anti-Ro (SSA) antibodies. This study shows that the dust-like particles staining pattern is not specific for SCLE, but is highly suggestive of connective tissue disease. The nature of the antigen responsible for the immunoglobulin deposition and the prognostic significance of this DIF pattern remain to be established. PMID- 9747371 TI - Annular lichen planus showing a change in metallothionein expression on immunohistochemistry. AB - The immunohistochemical localization of metallothioneins (MTs), low-molecular weight metal-binding proteins, was investigated in a case of annular lichen planus (LP) that enlarged centrifugally with healing in the centre after a month. The annular lesion was found to exhibit the early, developed and late stages of LP, as judged by the clinical and histopathological findings. Immunostaining for MTs was increased around the lesion, discontinuous around the rim of the erythema and undetectable in the centre of the lesion. MT expression was examined in eight specimens of idiopathic LP. Discontinuous immunostaining for MTs was observed in six specimens and increased staining around the lesion was observed in two specimens. These results suggest that MT expression changes depending on the inflammatory stage of LP. Although the role of MTs in dermatological disease is still unknown, their expression might be related to the pathological process of LP. PMID- 9747372 TI - Reticulolinear aplasia cutis congenita of the face and neck: a distinctive cutaneous manifestation in several syndromes linked to Xp22. AB - A distinct form of aplasia cutis congenita presenting as linear facial skin defects has been described under a variety of names as Xp deletion syndrome. MIDAS (microphthalmia, dermal aplasia and sclerocornea) syndrome, MLS (microphthalmia and linear skin defects) and Gazali-Temple syndrome. The syndrome is lethal in males, and its severity in females varies from a relatively mild residual facial scarring with short stature to lethal developmental organ malformations. A new case with peculiar ultrastructural findings is presented. A review of the literature suggests that these associations represent a series of contiguous-gene syndromes. PMID- 9747373 TI - Discoid lupus erythematosus associated with a primary immunodeficiency syndrome showing features of non-X-linked hyper-IgM syndrome. AB - Hyper-IgM (HIM) syndrome is a rare primary immunodeficiency disorder. Approximately 120 cases have been described in the literature world-wide. Features of HIM include low serum IgG, a very low IgA with normal or high IgM levels. Autoimmune phenomena are recognized associations but connective tissue disorders have so far not been described in HIM patients. We report the case of a 19-year-old Indian woman with an immunodeficiency syndrome characteristic of non X-linked HIM who developed discoid lupus erythematosus. Anti-double-stranded DNA antibodies were negative. Antibodies to extractable nuclear antigens were positive for Ro and nRNP, with evidence that they were of both IgG and IgM class. Treatment with hydroxychloroquine and topical steroids was successful. PMID- 9747374 TI - Recurrent annular erythema in juvenile chronic myelogenous leukaemia. AB - Juvenile chronic myelogenous leukaemia (JCML) is a rare haematological malignancy of myelomonocytic lineage that affects patients less than 4 years of age and is known as an entity different from adult-type chronic myelogenous leukaemia. In JCML, skin manifestations are relatively common but most of them have been reported as a non-specific eruption, which histologically may show changes resembling neurofibromatosis or xanthogranuloma. We present a 2-year-old boy with JCML who developed a recurrent annular erythema in which leukaemic infiltrates were confirmed histologically, even though his bone marrow examination suggested that be remained in haematological remission. PMID- 9747375 TI - Cutaneous vascular infarcts secondary to spontaneous platelet aggregation. AB - We report a 23-year-old woman presenting with multiple cutaneous infarcts who was found to have spontaneous platelet aggregation (SPA). Her skin lesions showed a good response to aspirin therapy and platelet aggregation returned to normal. Subsequently, she was found to have and was treated for pulmonary tuberculosis. The SPA may have been related to this via the formation of immune complexes. Isolated cutaneous infarcts have not previously been described in association with SPA. The role of spontaneous and increased platelet aggregation in skin disorders is discussed. PMID- 9747376 TI - Basal cell carcinoma with neuroid type nuclear palisading: a report of three cases. AB - Nuclear palisading is a characteristic feature which is typically seen in neural tumours such as neurilemmoma, and also in some other tumours. We report here three patients with basal cell carcinoma who showed histological patterns similar to nuclear palisading. To our knowledge, this is the first such case report in the medical literature; we apply the term 'neuroid-type nuclear palisading' to these cases. In our patients, the spindle-shaped tumour cells were tightly packed and the nuclei were arranged uniformly to form this rare feature. PMID- 9747377 TI - Mycosis fungoides presenting as keratosis lichenoides chronica. AB - We report a patient with a 17-year history of reticulated keratotic papules on the trunk and limbs, and telangiectatic eruption on the face, in whom the diagnosis of keratosis lichenoides chronica was first established. However, the biopsies showed an epidermotropic infiltrate of small irregular CD4+ lymphocytes, and detection of a T-cell clone in the lesions by polymerase chain reaction confirmed the diagnosis of mycosis fungoides. Thus, keratosis lichenoides chronica can be an unusual and potentially misleading presentation of mycosis fungoides. PMID- 9747378 TI - Actinic superficial folliculitis. AB - We report a 31-year-old man with sterile follicular pustules on the shoulders, trunk and arms, recurring every year within 48-72 h of the year's first exposure to the sun. Both clinical and histological characteristics match those of the condition first described in 1985 under the name actinic superficial folliculitis. We discuss possible relationships with other similar conditions. PMID- 9747379 TI - Follicular pustulous granuloma annulare. AB - We report a 73-year-old woman presenting with recurrent eruptions of generalized follicular pustules. Histological examination revealed several palisading necrobiotic granulomas with mucin deposits, with a perifollicular distribution. A dense neutrophilic infiltrate in the upper portion of affected follicular structures gave rise to pustulous lesions. Scaly papules and pseudovesiculous lesions over the palms with deeper necrobiotic granulomas involving sweat glands and epidermal perforation coexisted in some of the eruptions with generalized pustules. We propose the term follicular pustulous granuloma annulare for this peculiar form of granuloma annulare, which widens the clinicopathological spectrum of presentation of this disease. PMID- 9747380 TI - Surgical treatment of granuloma inguinale. AB - Granuloma inguinale is an indolent, progressive, ulcerative and granulomatous skin disease caused by Calymmatobacterium granulomatosis. It is generally treated with antibiotics. However, long-standing and complicated disease requires surgical treatment. Two patients with extensive and multiple perianal fistulas and abscesses unresponsive to medical treatment were managed with radical surgical resection. The first patient healed by primary intention, but a diverting colostomy was made for the second patient and the tissue defect was closed with a rotation flap. Follow-up at 4 years revealed the disappearance of the symptoms and the absence of recurrence in both patients. PMID- 9747381 TI - Human onychomycosis caused by Trichophyton equinum transmitted from a racehorse. AB - We report fingernail onychomycosis caused by Trichophyton equinum in a farmer who breeds racehorses. In addition to the thumbnail, T. equinum had infected one of the racehorses. Oral terbinafine cured the infection in the farmer. PMID- 9747382 TI - The association of HLA-DQ7 with bullous pemphigoid is restricted to men. AB - This study examines in detail the HLA associations of 74 patients (40 women and 34 men) with bullous pemphigoid (BP) and compares their immunogenetic profile with that of 604 unrelated control subjects (238 women and 366 men). Correlations were sought between HLA antigens and the various BP disease parameters investigated. The presence of milia was the only clinical or laboratory finding which was linked with a specific HLA antigen, HLA-DQ6, in both men and women with BP (P < 0.01). BP has previously been linked with the HLA-DQ7 antigen and this association was confirmed in 39 of our patients (14 women and 25 men). Twelve of these patients (four women and eight men) were homozygous for HLA-DQ7. The association of HLA-DQ7 with BP was gender-restricted and only significant for men (P < 0.01). No equivalent HLA disease susceptibility risk factor could be identified for our female BP patients. This difference in HLA association between men and women with BP has not been reported previously, and its significance for disease pathogenesis is not known. No specific link could be found between HLA DQ7 and BP for any of the clinical, immunofluorescence, western blotting, treatment or prognostic disease factors studied. PMID- 9747383 TI - Elevated soluble Fas levels in herpes zoster patients. PMID- 9747384 TI - Atopic eczema in monozygotic twins with Dubowitz syndrome. PMID- 9747385 TI - Mesothelioma-associated antiphospholipid antibody syndrome presenting with cutaneous infarction and neuropathy. PMID- 9747386 TI - Allergic contact dermatitis to acrylates in diathermy plates. PMID- 9747388 TI - Suction pads related to thumb sucking and chewing. PMID- 9747387 TI - Repetitive hair pulling associated with schizophrenia. PMID- 9747389 TI - Does ultraviolet radiation exposure influence S100 beta protein plasma levels? PMID- 9747390 TI - Malignant melanoma after psoralen and ultraviolet A (PUVA) therapy. PMID- 9747391 TI - Successful treatment of erythrodermic psoriasis with mycophenolate mofetil. PMID- 9747392 TI - Intravenous immunoglobulins for pemphigus vulgaris: adjuvant or first choice therapy? PMID- 9747393 TI - Subcutaneous sarcoidosis worsened by cyclosporin treatment for pyoderma gangrenosum. PMID- 9747394 TI - Disgust--the forgotten emotion of psychiatry. PMID- 9747396 TI - People with dementia can remember. Implications for care. PMID- 9747395 TI - Reserpine exhumed. PMID- 9747397 TI - Self-injury and violence in people with severe learning disabilities. AB - BACKGROUND: Psychiatry in severe and profound learning disability is essentially behavioural psychiatry. Some clinical and research observations of disorders of behaviour in this group are summarised in this study. METHOD: After inspection of the literature, I postulated a clinical syndrome of violence and self-injury in the severely learning disabled. A check-list of behavioural symptoms was developed and used in a community survey. RESULTS: Behaviour, assessed by the check-list, supported the existence of organic behaviour disorder, as did small scale psychophysiological testing. CONCLUSIONS: Self-injury is strongly associated with violence, and with severe and profound learning disability. Pathophysiology of violence and self-injury may include high levels of psychophysiological arousal demonstrated by unstable EEGs. Reduction of arousal by antipsychotic medication is associated with clinical improvement in violent and self-injurious behaviours. PMID- 9747398 TI - Self-injurious behaviour as part of genetic syndromes. AB - BACKGROUND: The purpose of this paper is to review the association between genetic syndromes and self-injurious behaviour. METHOD: The information available from the literature on the subject of self-injurious behaviours and genetic syndromes was collated and presented with a critical appraisal. RESULTS: Self injurious behaviours are associated with some genetic syndromes. However, the causal relationship between the genetic syndromes and the self-injurious behaviour remains far from clear. CONCLUSIONS: Although self-injurious behaviour has been shown to be the part of a broader phenotype in many genetic disorders, the specificity and sensitivity of these behaviours in this context remain unclear. PMID- 9747399 TI - Psychopharmacology of severe self-injury associated with learning disabilities. AB - BACKGROUND: Many sedative and antipsychotic agents have been used in the management of severe self-injury associated with learning disabilities. Their efficacy has been questioned. Recent research has identified some biological abnormalities associated with severe self-injury and allowed a more rational selection of treatment. METHOD: Review of published literature, including trials, previous reviews and case reports. REPORTS: There is evidence for the efficacy of opiate antagonists in the management of severe self-injury, and recent research has identified potential methods of predicting treatment response. Dopamine D1 antagonists and some agents affecting serotonin turnover may also be of benefit. CONCLUSIONS: More rational psychopharmacological treatments for severe self injurious behaviour may become available. Such treatments are difficult to evaluate for methodological and ethical reasons. They usually involve the clinical use of compounds for unlicensed indications, rather than trials of agents developed specifically to treat severe self-injurious behaviour. Combining psychopharmacological and psychological interventions may provide additional benefits. PMID- 9747400 TI - Psychological interventions in self-injurious behaviour. Working with people with a learning disability. AB - BACKGROUND: Psychological approaches to working with people with learning disabilities who self-injury have developed over the past 30 years. METHOD: The major literature is reviewed and an ecological framework is described which emphasises the importance of environmental, interpersonal and intrapersonal dynamics to understanding the multi-factorial nature of self-injury. Case examples are given. RESULTS: Self-injury is seen as essentially communicative and functionally adaptive; it is the person's best attempt to deal with abusive, neglecting or traumatic environments or events. CONCLUSIONS: The persistence of self-injurious behaviour once established, requires an interdisciplinary approach which addresses comprehensively the variety of factors which have contributed to the development and maintenance of self-injury. PMID- 9747401 TI - Auditing electroconvulsive therapy. The third cycle. AB - BACKGROUND: This is the third large-scale audit in the past 20 years and compares the practice of electroconvulsive therapy (ECT) in England and Wales with the standards derived from the Royal College of Psychiatrists' 2nd ECT handbook. METHOD: Facilities, equipment practice, personnel and training were systematically evaluated during visits to all ECT clinics in the former North East Thames and East Anglia regions and Wales. All other English ECT clinics were surveyed with a postal questionnaire. Information was obtained for 184 (84%) of the 220 ECT clinics identified. RESULTS: Although some aspects of ECT administration had improved since the last audit in 1991, overall only one-third of clinics were rated as meeting College standards. Only 16% of responsible consultants attended their ECT clinic weekly and only 6% had sessional time for ECT duties. Fifty-nine per cent of all clinics had machines of the type recommended by the College and 7% were still using machines considered outdated in 1989. Only about one-third of clinics had clear policies to help guide junior doctors to administer ECT effectively. CONCLUSIONS: Twenty years of activity by the Royal College of Psychiatrists and three large-scale audits have been associated with only modest improvement in local practice. PMID- 9747402 TI - Home self-assessment of obsessive-compulsive disorder. Use of a manual and a computer-conducted telephone interview: two UK-US studies. AB - BACKGROUND: Two studies tested whether subjects with obsessive-compulsive disorder could successfully use BT STEPS, a computer-aided system, to perform self-assessment for self-treatment of obsessive-compulsive disorder by exposure and ritual prevention. METHOD: Subjects were given a self-guiding manual and could use a touch-tone telephone to access computer-controlled Interactive Voice Response interviews at their convenience from home. Using the BT STEPS system, patients rated themselves and worked out a plan for individually tailored self exposure therapy. RESULTS: Outcomes were similar in the two studies. Of the 63 subjects who used BT STEPS, 84% completed the self-assessment module. Most calls were made outside usual office hours. As expected, subjects did not improve merely by completing self-assessment. However, completion of self-assessment predicted later improvement with self-exposure therapy. CONCLUSIONS: Most subjects successfully completed self-assessment using BT STEPS from their homes. DECLARATION OF INTEREST: BT STEPS is a trademark of Pfizer, Inc. I.M.M., L.B. and J.H.G. have a financial interest in BT STEPS. PMID- 9747403 TI - Randomised controlled trial of compliance therapy. 18-month follow-up. AB - BACKGROUND: A randomised controlled trial was conducted in an acute treatment setting to examine the effectiveness of compliance therapy, a brief pragmatic intervention targeting treatment adherence in psychotic disorders, based on motivational interviewing and recent cognitive approaches to psychosis. METHOD: Seventy-four patients with psychotic disorders according to DSM-III-R criteria recruited from consecutive admissions to an acute in-patient unit, received 4-6 sessions of either compliance therapy or non-specific counselling, and were followed-up over 18 months. The principal outcome measures were observer-rated compliance, attitudes to treatment, insight and social functioning. RESULTS: Significant advantages were found for the compliance therapy group post-treatment on measures of insight, attitudes to treatment and observer-rated compliance which were retained over the follow-up period. Global social functioning improved relatively more over time in the compliance therapy group compared with the control group. Survival in the community prior to readmission was significantly longer in the compliance therapy group. CONCLUSIONS: The results support the effectiveness of compliance therapy in improving functioning and community tenure after an acute psychotic episode. PMID- 9747405 TI - Amnestic people with Alzheimer's disease who remembered the Kobe earthquake. AB - BACKGROUND: Emotional memory is a special category of memory for events arousing strong emotions. To investigate the effects of emotional involvement on memory retention in individuals with Alzheimer's disease we studied peoples' memories of distressing experiences during a devastating earthquake. METHODS: Fifty-one subjects with probable Alzheimer's disease who experienced the Kobe earthquake at home in the greater Kobe area were studied. Memories of the earthquake were assessed 6 and 10 weeks after the disaster in semi-structured interviews, and were compared with memories of a magnetic resonance imaging (MRI) examination given after the earthquake. RESULTS: Forty-four (86.3%) of the subjects remembered the earthquake and 16 (31.4%) of subjects remembered the MRI experience. Factual content of the earthquake was lost in most of the subjects. CONCLUSIONS: Fear reinforces memory retention of an episode in subjects with Alzheimer's disease but does not enhance retention of its context, despite repeated exposure to the information. PMID- 9747404 TI - Cost-effectiveness evaluation of compliance therapy for people with psychosis. AB - BACKGROUND: Non-compliance rates with antipsychotic medication can be high, and the personal and societal costs are considerable. A new psychological intervention, compliance therapy seeks to improve compliance and patient outcomes and reduce treatment costs. METHOD: A randomised controlled study examined the cost-effectiveness of compliance therapy compared to non-specific counselling over 18 months for 74 people with psychosis admitted as inpatients at the Maudsley Hospital. Bivariate and multivariate analyses were conducted to test for differences and to explore inter-patient cost variations. RESULTS: Compliance therapy is more effective and is no more expensive. Consequently, compliance therapy is more cost-effective than non-specific counselling at six, 12 and 18 months. CONCLUSIONS: There are compliance, outcome and cost-effectiveness arguments in favour of compliance therapy in preference to non-specific counselling. PMID- 9747406 TI - Epidemiology of paranoid symptoms in an elderly population. AB - BACKGROUND: Elderly people with paranoid symptoms are a taxing group for medical and social services, but studies of the prevalence of these symptoms in the general elderly population are rare. This study aimed to estimate the community prevalence and to identify some associated variables. METHOD: A community samples of 1420 elderly people, was extensively examined by nurses and physicians. RESULTS: Paranoid ideation was found in 6.3% of the sample. The prevalence in people with cognitive dysfunction (n = 381, 12.1%) was higher than in those without (n = 1039, 2.6%). Once cognitive impairment had been controlled the associated variables were: being divorced, being female, having depressive symptoms, using psychotropic drugs, having no friends or visitors, using community care and being an immigrant. CONCLUSION: Paranoid symptoms in this elderly population were associated most strongly with cognitive impairment. Other associated variables pointed to a higher level of social isolation than others in the community. PMID- 9747407 TI - Detecting postnatal depression in Chinese women. Validation of the Chinese version of the Edinburgh Postnatal Depression Scale. AB - BACKGROUND: We evaluated the utility of the Chinese version of the Edinburgh Postnatal Depression Scale (EPDS) and measured the prevalence of major depression six weeks after confinement among Chinese women in Hong Kong. METHOD: A prospective cohort of 145 women completed the EPDS, the 12-item General Health Questionnaire (GHQ) and the Beck Depression Inventory (BDI) six weeks after giving birth. They were then assessed with the Structured Clinical Interview for DSM-III-R, non-patient version (SCID-NP) to establish psychiatric diagnosis. The criterion validity of EPDS was tested against this clinical diagnosis, and the concurrent validity against the GHQ and BDI scores was also evaluated. The internal consistency of the scales was measured by Cronbach's alpha coefficient. RESULTS: The Chinese EPDS had satisfactory psychometric properties and a cut-off score of 9/10 is recommended for screening depressive illness in a general postnatal population. At six weeks postpartum, 5.5% of the study population suffered from major depression. CONCLUSIONS: The Chinese EPDS will be useful for screening for postnatal depression. PMID- 9747408 TI - Prolactin response to d-fenfluramine is blunted in people with anorexia nervosa. AB - BACKGROUND: Several studies have explored serotonin (5-HT) transmission in people with anorexia nervosa, but their results have been inconsistent. METHOD: According to a double-blind placebo-controlled design, plasma prolactin response to the specific serotonergic probed d-fenfluramine was investigated in 10 under weight and two normal-weight women with anorexia, and in 12 age-matched healthy females. Eating-related psychopathology, depressive and obsessive--compulsive symptoms, and aggressiveness were measured by appropriate rating scales. RESULTS: Compared with healthy control subjects, the women with anorexia showed reduced baseline prolactin and oestrogen levels and increased basal cortisol concentrations. The prolactin response to d-fenfluramine was blunted and did not correlate with psychopathological measures. CONCLUSIONS: These results support a dysfunction of 5-HT transmission in anorexia nervosa. This dysfunction does not seem to be related to concomitant depressive or obsessive--compulsive symptoms or to the level of aggressiveness of the patients. PMID- 9747409 TI - Post-traumatic stress disorder in children and adolescents following road traffic accidents. AB - BACKGROUND: Post-traumatic stress disorder (PTSD) can be a persistent and disabling psychiatric disorder. There is little systematic research into the psychiatric consequences of road traffic accidents (RTAs) in children and adolescents. METHOD: A consecutive sample of 8-16-year-olds attending an accident and emergency department following RTAs were screened for PTSD. Potential cases and their parent(s) were interviewed with semi-structured research instruments about six weeks and six months after the accident. RESULTS: Fifty-three (45%) of the 119 subjects fell above PTSD cut-off on the Frederick's Reaction Index. Thirty-three (75%) of the 44 cases met DSM-IV criteria for PTSD. In half of these other psychiatric disorders were present, including major depressive disorder and anxiety disorders. Being female, involvement in car accidents and pre-existing depression and anxiety were associated with developing PTSD. Seventeen per cent of the sample continued to be symptomatic six months after the accident. CONCLUSIONS: PTSD is a common consequence of RTAs. Liaison with accident and emergency departments would enhance the early detection and follow-up of children at risk of developing PTSD. PMID- 9747410 TI - Treatment of severe personality disorder. PMID- 9747411 TI - Economics of attachment disorders. PMID- 9747412 TI - PTSD and victims of torture. PMID- 9747413 TI - Cost-effective community psychiatry. PMID- 9747414 TI - Sulpiride augmentation on schizophrenia. PMID- 9747415 TI - Amisulpride in schizophrenia. PMID- 9747416 TI - Systematic does not necessarily mean comprehensive. PMID- 9747417 TI - What counts as clinical research? PMID- 9747418 TI - Rapid intravenous detoxification in heroin addiction. PMID- 9747419 TI - Moclobemide in social phobia. PMID- 9747420 TI - Liaison between adolescent and adult services in early-onset schizophrenia. PMID- 9747422 TI - Can transsexualism remit? PMID- 9747421 TI - Somatoform dissociation is unlikely to be a result of indoctrination by therapists. PMID- 9747423 TI - The membrane current (I(f)) in human atrial cells: implications for atrial arrhythmias. PMID- 9747424 TI - Once too little, now too much. PMID- 9747425 TI - Evolution and clinical implications of the myofibroblast concept. PMID- 9747426 TI - "Myocardial stunning" remaining questions. PMID- 9747427 TI - Cardiac protein phosphorylation: functional and pathophysiological correlates. AB - Protein phosphorylation acts a pivotal mechanism in regulating the contractile state of the heart by modulating particular levels of autonomic control on cardiac force/length relationships. Early studies of changes in cardiac protein phosphorylation focused on key components of the excitation-coupling process, namely phospholamban of the sarcoplasmic reticulum and myofibrillar troponin I. In more recent years the emphasis has shifted towards the identification of other phosphoproteins, and more importantly, the delineation of the mechanistic and signaling pathways regulating the various known phosphoproteins. In addition to cAMP- and Ca(2+)-calmodulin-dependent kinase processes, these have included regulation by protein kinase C and the ever-emerging family of growth factor related kinases such as the tyrosine-, mitogen- and stress-activated protein kinases. Similarly, the role of protein dephosphorylation by protein phosphatases has been recognized as integral in modulating normal cardiac cellular function. Recent studies involving a variety of cardiovascular pathologies have demonstrated that changes in the phosphorylation states of key cardiac regulatory proteins may underlie cardiac dysfunction in disease states. The emphasis of this comprehensive review will be on discussing the role of cardiac phosphoproteins in regulating myocardial function and pathophysiology based not only on in vitro data, but more importantly, from ex vivo experiments with corroborative physiological and biochemical evidence. PMID- 9747428 TI - The control of Ca release from the cardiac sarcoplasmic reticulum: regulation versus autoregulation. AB - This review discusses the mechanism and regulation of Ca release from the cardiac sarcoplasmic reticulum. Ca is released through the Ca release channel or ryanodine receptor (RyR) by the process of calcium-induced Ca release (CICR). The trigger for this release is the L-type Ca current with a small contribution from Ca entry on the Na-Ca exchange. Recent work has shown that CICR is controlled at the level of small, local domains consisting of one or a small number of L-type Ca channels and associated RyRs. Ca efflux from the s.r. in one such unit is seen as a 'spark' and the properties of these sparks produce controlled Ca release from the s.r. A major factor controlling the amount of Ca released from the s.r. and therefore the magnitude of the systolic Ca transient is its Ca content. The Ca content depends on both the properties of the s.r. and the cytoplasmic Ca concentration. Changes of s.r. Ca content and the Ca released affect the sarcolemmal Ca and Na-Ca exchange currents and this acts to control cell Ca loading and the s.r. Ca content. The opening probability of the RyR can be regulated by various physiological mediators as well as pharmacological compounds. However, it is shown that, due to compensatory changes of s.r. Ca, modifiers of the RyR only produce transient effects on systolic Ca. We conclude that, although the RyR can be regulated, of much greater importance to the control of Ca efflux from the s.r. are effects due to changes of s.r. Ca content. PMID- 9747429 TI - Fifteen years experience with finger arterial pressure monitoring: assessment of the technology. AB - We review the Finapres technology, embodied in several TNO-prototypes and in the Ohmeda 2300 and 2300e Finapres NIBP. Finapres is an acronym for FINger Arterial PRESsure, the device delivers a continuous finger arterial pressure waveform. Many papers report on the accuracy of the device in comparison with intra arterial or with noninvasive but intermittent blood pressure measurements. We compiled the results of 43 such papers and found systolic, diastolic and mean accuracies, in this order, ranging from -48 to 30 mmHg, from -20 to 18 mmHg, and from -13 to 25 mmHg. Weighted for the number of subjects included pooled accuracies were -0.8 (SD 11.9), -1.6 (8.3) and -1.6 (7.6) mmHg respectively. Subdividing the pooled group according to criteria such as reference blood pressure, place of application, and prototype or commercial device we found no significant differences in mean differences or SD. Measurement at the finger allows uninterrupted recordings of long duration. The transmission of the pressure pulse along the arm arteries, however, causes distortion of the pulse waveform and depression of the mean blood pressure level. These effects can be reduced by appropriate filtering, and upper arm 'return-to-flow' calibration to bring accuracy and precision within AAMI limits. For the assessment of beat-to beat changes in blood pressure and assessment of blood pressure variability Finapres proved a reliable alternative for invasive measurements when mean and diastolic pressures are concerned. Differences in systolic pressure are larger and reach statistical significance but are not of clinical relevance. Finger arteries are affected by contraction and dilatation in relation to psychological and physical (heat, cold, blood loss, orthostasis) stress. Effects of these phenomena are reduced by the built-in Physiocal algorithm. However, full smooth muscle contraction should be avoided in the awake patient by comforting the patient, and covering the hand. Arterial state can be monitored by observing the behaviour of the Physiocal algorithm. We conclude that Finapres accuracy and precision usually suffice for reliable tracking of changes in blood pressure. Diagnostic accuracy may be achieved with future application of corrective measures. PMID- 9747430 TI - Polymorphic ventricular tachycardias induced by D-sotalol and phenylephrine in canine preparations of atrioventricular block: initiation in the conduction system followed by spatially unstable re-entry. AB - OBJECTIVE: Polymorphic ventricular tachycardias (PVT) occur spontaneously in canine hearts under the combination of D-sotalol (S), bradycardia and phenylephrine (PE). We investigated the hypotheses that: (1) the activation patterns of the initial PVT beats would be consistent with an origin in the ventricular conduction system; and (2) the inhomogeneous prolongation of repolarisation intervals can provide refractory barriers for re-entrant activity. METHODS: Unipolar electrograms were recorded from 127 epicardial (EPI) sites with a sock electrode array as well as from intramural and endocardial sites during PVTs. Electrograms were analysed to generate isochronal maps and measure the spatial distribution of activation-recovery intervals (ARI). RESULTS: Under S (9.9-14.5 mg.l-1), spontaneously terminating PVTs (cycle length of 270 +/- 43 ms, n = 45) (mean +/- s.d.) occurred when a PE bolus (10-50 micrograms.kg-1) was injected. The first beat of the PVTs occurred with a coupling interval of several hundred ms to the preceding idioventricular beat (IDV) without any bridging activity and its earliest EPI breakthrough occurred in areas overlying the terminations of the right or left bundle branch. ARI values measured in IDV (295 +/- 47 ms) were significantly prolonged prior to PVT (462 +/- 92 ms). Prolongation was greater in apical than in basal epicardial areas, and at endocardial than epicardial sites (to > 500 ms). Maximum delays > 200 ms developed in the regions of marked ARI prolongation and, in later beats, circus movement re-entry occurred around refractory barriers, shifting between various regions of the ventricles. CONCLUSION: Thus, PVTs occurring spontaneously under conditions of delayed repolarisation originate from shifting sites in the ventricular conduction system and re-entrant activity shifting between various regions of the ventricle may occur in later beats of the more sustained arrhythmias. PMID- 9747431 TI - Angiotensin converting enzyme inhibition, AT1 receptor inhibition, and combination therapy with pacing induced heart failure: effects on left ventricular performance and regional blood flow patterns. AB - BACKGROUND: AT1 receptor activation has been demonstrated to cause increased vascular resistance properties which may be of particular importance in the setting of congestive heart failure (CHF). The overall goal of this study was to examine the effects of ACE inhibition (ACEI) alone, AT1 receptor blockade alone and combined ACEI and AT1 receptor blockade on LV pump function, systemic hemodynamics and regional blood flow patterns in the normal state and with the development of pacing induced CHF, both at rest and with treadmill induced exercise. METHODS AND RESULTS: Pigs (25 kg) were instrumented in order to measure cardiac output (CO), systemic (SVR) and pulmonary vascular (PVR) resistance, neurohormonal system activity, and myocardial blood flow distribution in the conscious state and assigned to one of 4 groups: (1) rapid atrial pacing (240 bpm) for 3 weeks (n = 7); (2) ACEI (benazeprilat, 3.75 mg/day) and pacing (n = 7); (3) AT1 receptor blockade (valsartan, 60 mg/day) and rapid pacing (n = 7); and (4) ACEI and AT1 receptor blockade (benazeprilat/valsartan, 1/60 mg/day, respectively) and pacing (n = 7). Measurements were obtained at rest and with treadmill exercise (15 degrees, 3 miles/h; 10 min) in the normal control state and after the completion of the treatment protocols. With rapid pacing, CO was reduced at rest and with exercise compared to controls. ACEI or AT1 blockade normalized CO at rest, but remained lower than control values with exercise. Combination therapy normalized CO both at rest and with exercise. Resting SVR in the CHF group was higher than controls and SVR fell to a similar degree with exercise; all treatment groups reduced resting SVR. With exercise, SVR was reduced from rapid pacing values in the ACEI and combination therapy groups. PVR increased by over 4-fold in the rapid pacing group both at rest and with exercise, and was reduced in all treatment groups. In the combination therapy group, PVR was similar to control values with exercise. Plasma catecholamines and endothelin levels were increased by over 3-fold with chronic rapid pacing, and were reduced in all treatment groups. In the combination therapy group, the relative increase in catecholamines and endothelin with exercise were significantly blunted when compared to rapid pacing only values. LV myocardial blood flow at rest was reduced in the rapid pacing only and monotherapy groups, but was normalized with combination therapy. CONCLUSION: These findings suggest that with developing CHF, combined ACE inhibition and AT1 receptor blockade improved vascular resistive properties and regional blood flow distribution to a greater degree than that of either treatment alone. Thus, combined ACEI and AT1 receptor blockade may provide unique benefits in the setting of CHF. PMID- 9747432 TI - The cardiac adrenergic system in ischaemia: differential role of acidosis and energy depletion. AB - OBJECTIVE: Acute myocardial ischaemia has been shown to modulate the beta adrenergic system and to activate protein kinase C. The aim of this study was to investigate if two important components of ischaemia, i.e. energy depletion or acidosis, may contribute to these changes. METHODS: Isolated rat hearts were perfused either with anoxia (in the absence of oxygen) or with cyanide in the absence of glucose as models of energy depletion with a loss of high energy phosphates. Alternatively, isolated hearts were perfused with acidic modified Krebs-Henseleit solution to induce acidosis. RESULTS: Energy depletion induced by cyanide perfusion leads to an increase of beta-adrenergic receptors (81 +/- 7 vs. 50 +/- 3 fmol/mg protein, p < or = 0.05) comparable to the changes observed in ischaemia, yet without any change of total adenylyl cyclase activity or protein kinase C activity. Similar, yet less pronounced changes were induced by anoxic perfusion. Acidic perfusion, in contrast, promotes a translocation of protein kinase C to the plasma membranes, suggesting its rapid activation. Additionally, an increased total forskolin-stimulated activity of adenylyl cyclase (515 +/- 16 vs. 428 +/- 17 pmol/min/mg, p < or = 0.05) was observed. Both were comparable to the sensitization observed in early ischaemia. In acidosis, the density of beta adrenergic receptors remained unaltered. CONCLUSIONS: These data suggest that the regulation of cardiac beta-adrenergic receptors is susceptible to energy depletion, but not to acidosis, whereas the intracellular enzymes both adenylyl cyclase and protein kinase C may be regulated by intracellular acidosis. This is the first differentiation of distinct components of ischaemia modulating the beta adrenergic signal transduction pathway. Both components may be operative in concert in acute myocardial ischaemia and may contribute to the regulation of these components of signal transduction observed in acute ischaemia. PMID- 9747433 TI - Effects of vasodilators and perfusion pressure on coronary flow and simultaneous release of nitric oxide from guinea pig isolated hearts. AB - OBJECTIVE: The aims were to validate the use of a direct reading NO electrode, to compare the effects of diverse acting drugs on altering coronary flow (CF) and NO release, and to examine the effects of altered perfusion pressure on flow-induced changes in NO concentration [NO] in the hemoglobin free effluent of guinea pig isolated hearts. METHODS: Hearts were isolated and perfused initially at a constant perfusion pressure (55 mmHg) with a modified Krebs-Ringer's solution equilibrated with 97% O2 and 3% CO2 at 37 degrees C. Heart rate, left ventricular pressure, CF, and effluent pH, pCO2, pO2, and NO generated current were monitored continuously on-line. Effluent was sampled for L-citrulline. Percent O2 extraction and O2 consumption were calculated. [NO] was quantitated with a sensitive amperometric sensor (sensitivity > or = 1 nmol/l approximately 3 pA) and a selective gas permeable membrane. RESULTS: The electrode was not sensitive to changes in solution pO2, flow, or pressure. The electrode was sensitive to pCO2 (-0.50 nmol/l/mmHg) and temperature (+24.5 nmol/l/degree C), so coronary effluent pCO2 was measured to compensate for a small decrease in pCO2 that occurred with an increase in coronary flow, and effluent temperature was rigidly controlled. Serotonin, bradykinin, and nitroprusside increased NO release along with CF, whereas nifedipine, butanedione monoxime, zaprinast, and bimakalim comparably increased CF but did not increase [NO] or NO release. Increases in CF (ml/g/min) and NO release (pmol/g/min), respectively, were 5.0 +/- 1 and 100 +/- 17 for 1 mumol/l serotonin, 7.5 +/- 1 and 148 +/- 18 for 100 nmol/l bradykinin, and 7.8 +/- 1 and 173 +/- 28 for 100 mumol/l nitroprusside. The increases in effluent NO by bradykinin were proportional to the increases in L-citrulline. Tetraethylammonium decreased CF, but did not change NO release, indomethacin changed neither CF nor NO release, and NG-nitro-L-arginine methyl ester (L-NAME) reduced CF by 2.6 +/- 1 ml/g/min and NO release by 25 +/- 8 pmol/g/min. An increase of CF of 8.0 +/- 0.3 ml/g/min, produced by increasing perfusion pressure from 25 to 90 mmHg, increased [NO] by 30 +/- 4 nmol/l; L-NAME but did not reduce the pressure-induced increase in CF, but reduced the increase in [NO] to 10 +/- 5 nmol/l. CONCLUSIONS: This study demonstrates in intact hearts real-time release of NO by several vasodilator drugs and by pressure-induced increases in flow (shear stress) and attenuation of these effects by L-NAME. PMID- 9747434 TI - The effect of changing excitation frequency on parallel conductance in different sized hearts. AB - OBJECTIVE: An important component of the ventricular volume measured using the conductance catheter technique is due to parallel conductance (Vc), which results from the extension of the electric field beyond the ventricular blood pool. Parallel conductance volume is normally estimated using the saline dilution method (Vc(saline dilution)), in which the conductivity of blood in the ventricle is transiently increased by injection of hypertonic saline. A simpler alternative has been reported by Gawne et al. [12]. Vc(dual frequency) is estimated from the difference in total conductance measured at two exciting frequencies and the method is based on the assumption that parallel conductance is mainly capacitive and hence is negligible at low frequency. The objective of this study was to determine whether the dual frequency technique could be used to substitute the saline dilution method to estimate Vc in different sized hearts. METHODS: The accuracy and linearity of a custom-built conductance catheter (CC) system was initially assessed in vitro. Subsequently, a CC and micromanometer were inserted into the left ventricle of seven 5 kg pigs (group 1) and six 50 kg pigs (group 2). Cardiac output was determined using thermodilution (group 1) and an ultrasonic flow probe (group 2) from which the slope coefficient (alpha) was determined. Steady state measurements and Vc estimated using saline dilution were performed at frequencies in the range of 5-40 kHz. All measurements were made at end-expiration. Finally, Vc was estimated from the change in end-systolic conductance between 5 kHz and 40 kHz using the dual frequency technique of Gawne et al. [12]. RESULTS: There was no change in measured volume of a simple insulated cylindrical model when the stimulating frequency was varied from 5-40 kHz. Vc(saline dilution) varied significantly with frequency in group 1 (8.63 +/- 2.74 ml at 5 kHz; 11.51 +/- 2.65 ml at 40 kHz) (p = 0.01). Similar results were obtained in group 2 (69.43 +/- 27.76 ml at 5 kHz; 101.24 +/- 15.21 ml at 40 kHz) (p < 0.001). However, the data indicate that the resistive component of the parallel conductance is substantial (Vc at 0 Hz estimated as 8.01 ml in group 1 and 62.3 ml in group 2). There was an increase in alpha with frequency in both groups but this did not reach significance. The correspondence between Vc(dual frequency) and Vc(saline dilution) methods was poor (group 1 R2 = 0.69; group 2 R2 = 0.22). CONCLUSION: At a lower excitation frequency of 5 kHz a smaller percentage of the electric current extends beyond the blood pool so parallel conductance is reduced. While parallel conductance is frequency dependent, it has a substantial resistive component. The dual frequency method is based on the assumption that parallel conductance is negligible at low frequencies and this is clearly not the case. The results of this study confirm that the dual frequency technique cannot be used to substitute the saline dilution technique. PMID- 9747435 TI - Cyclosporine A limits myocardial infarct size even when administered after onset of ischemia. AB - OBJECTIVE: The role of the immunosuppressant cyclosporine A as a preconditioning mimetic in the rabbit heart was examined. METHODS: Cyclosporine A, a potent protein 2B or calcium/calmodulin-dependent phosphatase (PP) inhibitor, was administered isolated rabbit hearts starting either 15 min prior to or 10 or 20 min after the onset of a 30 min period of regional ischemia and continuing until the onset of reperfusion. The effect of pretreatment with a second PP2B antagonist, FK-506, was also examined. In an additional protocol L-NAME was perfused for 50 min starting 5 min before the 45-min infusion of cyclosporine A. After 2 h of reperfusion infarct size was measured with triphenyltetrazolium chloride. In a second study left ventricular biopsies of isolated rabbit hearts were obtained to measure the effect of cyclosporine A on dephosphorylation of [32P] phosphorylase kinase by calcium/calmodulin-dependent phosphatases. RESULTS: Pretreatment with cyclosporine A resulted in only 10.0%, infarction of the risk zone, significantly less than that in untreated control hearts (28.7%, p < 0.001) but comparable to the extent of infarction in ischemically preconditioned hearts (10.0% p < 0.001 vs. control). Equivalent protection was also observed in hearts with treatment delayed for 10 min following the onset of ischemia (10.4% infarction, p < 0.001 vs. control). However, protection waned when cyclosporine A was administered only during the last 10 min of the 30-min ischemic period (25.5% infarction, p = n.s. vs. control). Pretreatment with FK-506 also resulted in myocardial salvage (10.4% infarction, p < 0.001 vs. control). When hearts were exposed to a co-infusion of L-NAME and cyclosporine A, protection was still evident (18.1% infarction, p < 0.05 vs. L-NAME), although not as robust as that seen with the PP2B blocker alone. In hearts pretreated with cyclosporine A dephosphorylation of [32P] phosphorylase kinase by calcium/calmodulin-dependent phosphatases was inhibited by 67%. CONCLUSIONS: Cyclosporine A and FK-506, potent PP2B inhibitors, can protect the ischemic rabbit heart, and at least cyclosporine A continues to be effective when infusion is delayed until after the onset of ischemia. The mechanism of this protection may be related to inhibition of phosphatases and prolongation of the phosphorylation state of ischemic cells. PMID- 9747436 TI - Preferential inhibition of Ikr by MCI-154, a putative cardiotonic Ca2+ sensitizer, in guinea pig atrial cells. AB - OBJECTIVE: To define the electrophysiologic mechanism(s) by which MCI-154, a putative Ca2+ sensitizer, produces a positive inotropic response without a positive chronotropic response, we examined effects of MCI-154 on the action potential of atrial preparations and the membrane currents of atrial myocytes. METHODS: The action potentias were recorded from left atrial and sinoatrial node preparations of guinea pigs by the use of standard microelectrode techniques. The whole-cell membrane currents were recorded from enzymatically-dissociated guinea pig atrial myocytes using conventional patch clamp techniques. RESULTS: In isolated left atria, MCI-154 increased the developed tension in a concentration dependent manner. MCI-154 at concentrations of 10 and 100 microM increased the action potential duration (APD) in left atria stimulated at 0.5 Hz. In sinoatrial node preparations MCI-154 at a concentration of 100 microM produced a negative chronotropic response and prolonged APD. In single right atrial myocytes, MCI-154 at concentrations of 10 and 100 microM failed to increase the inward L-type Ca2+ current, but decreased the delayed rectifier K+ current (IK) in a concentration dependent manner. MCI-154 decreased IK elicited by short depolarizing pulses more markedly than that induced by long depolarizing pulses. In addition, MCI-154 produced only a little inhibition of IK in the presence of E-4031, a specific blocker of rapidly activating component of IK (IKr). CONCLUSIONS: MCI-154 preferentially blocks IKr and the inhibitory action on IKr may be partly involved in the negative chronotropic and positive inotropic responses in atrial preparations. PMID- 9747437 TI - Free radical involvement in doxorubicin-induced electrophysiological alterations in rat papillary muscle fibres. AB - OBJECTIVE: This work was designed to determine whether the doxorubicin-induced changes in heart electrical activity are due to increased free radical production and membrane oxidative damage. METHODS: Four groups of rats (60 days old) were used. One group was untreated and the others were treated with doxorubicin (DXR), DXR and vitamin E, and DXR and N-acetylcysteine (NAC), respectively. DXR was administered by single i.p. injection (20 mg/kg b.wt.). Vitamin E was administered by ten daily i.m. injections (100 mg/kg), while NAC (100 mg/kg) was injected i.p. 1 h before and 7 h after DXR. The effectiveness of the drug in inducing oxidative stress in different tissues and of the antioxidants in offering protection was established by determining antioxidant capacity, susceptibility to oxidative stress, and lipid peroxidation in heart, liver, and blood. The drug effect on heart electrical activity was determined by measuring the heart rate in vivo and action potential configuration in papillary muscle fibres in vitro. Heart lipid peroxidation and electrical activity were also examined in both vitamin E and NAC-treated rats. RESULTS: DXR treatment decreased antioxidant capacity and increased lipid peroxidation and susceptibility to oxidative stress in heart and blood, but not in liver. DXR administration to rats treated with antioxidants did not produce significant changes in antioxidant capacity and susceptibility to oxidative stress even in heart and blood. Furthermore, lipid peroxidation in heart and liver from DXR- and vitamin E treated rats, and in liver from DXR- and NAC-treated rats was lower than in untreated controls. DXR treatment also increased the duration of ventricular action potentials in untreated rats, but not in antioxidant-treated rats. The treatment of control animals with the antioxidants affected lipid peroxidation, but not cardiac electrical activity. CONCLUSIONS: The protection offered by antioxidants against electrophysiological alterations indicates a free radical involvement in such alterations. In contrast, although electrical modifications are associated with increased peroxidative processes and both are prevented by the antioxidants, it is not yet clear whether a causative relationship exists between them. PMID- 9747438 TI - Phenylephrine-induced stimulation of Na+/Ca2+ exchange in rat ventricular myocytes. AB - OBJECTIVE: The effect of an alpha-adrenergic agonist, phenylephrine, on the Na+/Ca2+ exchange current in rat ventricular myocytes was investigated. METHODS: The Na+/Ca2+ exchange current was measured at room temperature in rat ventricular myocytes as the whole-cell current induced by addition of extracellular Na+ and Ca2+, while blocking Na+ current by setting the holding potential at -30 mV, K+ currents by intracellular Cs+, TEA+ and by extracellular Ba2+, Ca2+ current by nifedipine and Na+ pump current by ouabain or by 0 extracellular K+. RESULTS: Under these experimental conditions, application of external Na+ and Ca2+ induced a current which was further increased by phenylephrine. Phenylephrine (80 microM) increased the current by up to 31.0 +/- 5.4% of control at all membrane potentials tested both below and above the reversal potential. The reversal potential (+21.0 +/- 3.2 mV), which corresponded with the theoretical reversal potential for the Na+/Ca2+ exchange current under our ionic conditions (+21.3 mV), was not changed by phenylephrine (+23.2 +/- 4.1 mV). Applying phenylephrine in the absence of Na+/Ca2+ exchange (0 Na+e, 0 Ca2+e) did not change the current. The effect was resistant to propranolol, a beta-adrenergic blocker, but prevented by prazosin, an alpha-receptor antagonist, by neomycin, an inhibitor of phospholipase C, and by chelerythrine, a selective inhibitor of protein kinase C. Phorbol 12-myristate 13-acetate failed to stimulate the current. The effect remained similar under conditions of high (HEPESi = 5 mM) and low (HEPESi = 0.5 mM) intracellular pH buffering. CONCLUSION: Our data indicate that phenylephrine stimulates the Na+/Ca2+ exchange, both in the forward and the reverse modes, probably via a protein kinase C-dependent pathway. PMID- 9747440 TI - Selective dysregulation of nitric oxide synthase type 3 in cardiac myocytes but not coronary microvascular endothelial cells of spontaneously hypertensive rat. AB - OBJECTIVE: Recent studies indicate that endothelial type nitric oxide synthase (NOS3) modulates cardiac systolic and diastolic function and the inotropic responsiveness to beta-adrenergic agonists, and may affect myocardial oxygen consumption. Although NOS3 is a constitutive protein, its levels of expression can be modified by various physiological and pathophysiological stimuli. We investigated whether the cell-specific expression of NOS3 mRNA and protein are altered in cardiac hypertrophy. METHODS: Left ventricular cardiac myocytes and coronary microvascular endothelial cells were freshly isolated from 12 week old male spontaneously hypertensive rat (SHR) and matched normotensive Wistar rat hearts. NOS3 protein levels were assessed by Western analysis, and mRNA levels by RT-PCR and Southern blotting. RESULTS: Left ventricular/body weight ratios were significantly increased in SHR compared to Wistar controls, indicating significant hypertrophy. The levels of NOS3 protein were markedly decreased in SHR compared to Wistar cardiac myocytes (by approximately 85%). By contrast, the expression of NOS3 mRNA normalized for GAPDH was increased approximately 3 fold in SHR cardiac myocytes relative to Wistar controls. In freshly isolated microvascular endothelial cells, however, levels of NOS3 protein and NOS3 mRNA were similar between the two groups. CONCLUSIONS: The expression of NOS3 is selectively altered in cardiac myocytes but not coronary microvascular endothelial cells of young SHR hearts, with a marked decrease in NOS3 protein but an increase in NOS3 mRNA. This dysregulation of NOS3 could contribute to contractile dysfunction in left ventricular hypertrophy. PMID- 9747439 TI - Proteolysis of connexin43-containing gap junctions in normal and heat-stressed cardiac myocytes. AB - OBJECTIVE: The present studies were performed to examine the degradation of connexin43-containing gap junctions by the lysosome or the proteasome in normal and heat-stressed cultures of neonatal rat ventricular myocytes. METHODS: Primary cultures were prepared from neonatal rat ventricular myocytes. Connexin43 was detected by immunoblotting, immunofluorescence, or immunoprecipitation. Gap junction profiles were detected by transmission electron microscopy. RESULTS: Immunoblots of whole cell lysates demonstrated increased levels of connexin43 in cultures treated with lysosomal inhibitors (chloroquine, leupeptin, E-64, or ammonium chloride) or proteasomal inhibitors (lactacystin or ALLN). Pulse-chase experiments showed that the half-life of connexin43 was 1.4 h in control cultures, but was prolonged to 2.0 or 2.8 h in cultures treated with chloroquine or lactacystin, respectively. Immunofluorescence and electron microscopy showed a significant increase in the number of gap junction profiles in myocytes treated with either chloroquine or lactacystin. Heat treatment of cultures (43.5 degrees C for 30 min) produced a rapid loss of connexin43 as detected by immunoblotting or immunofluorescence. Heat-induced connexin43 degradation was prevented by simultaneous treatment with lactacystin, ALLN, or chloroquine. Connexin43 levels and distribution returned to normal by 3 h following a heat shock and were resistant to a subsequent repeat heat stress. The heat shock also led to production of HSP70 as detected by immunoblotting. CONCLUSIONS: These data suggest that Cx43 gap junctions in myocytes are degraded by the proteasome and the lysosome, that this proteolysis can be augmented by heat stress, and that inducible factors such as HSP70 may protect against Cx43 degradation. PMID- 9747441 TI - Super-normal 201Tl retention in hibernating myocardium: an ex-vivo study using the failing human heart. AB - OBJECTIVE: Although the relationship between delayed 201Tl distribution and blood flow in acutely ischemic and infarcted myocardium has been widely explored in the experimental setting, its behaviour in chronically hypoperfused dysfunctioning human myocardium has not yet been evaluated. METHODS: In tissue samples of excised failing hearts taken from ischemic (IHD) patients and idiopathic dilated cardiomyopathy (IDC) controls, we evaluated the relationship between delayed 201Tl retention (4 h redistribution), blood flow (assessed by means of 99mTc labelled human albumin microspheres injected during transplantation) and biochemically-assessed fibrosis. 201Tl activity was expressed as the percent of the activity in the region with highest flow and the least fibrosis. RESULTS: Fibrosis and 201Tl activity were inversely related (r = -0.62, P = 0.0001). In IDC controls, low flows corresponded to uniformly preserved 201Tl retention. In IHD, 46 segments with flows < or = 0.60 ml.min-1.g-1 and 20 segments with flows > 0.60 ml.min-1.g1 showed matching delayed 201Tl retention and flow values; in the remaining 27, there was a disproportionately high tracer accumulation in comparison with flow (flow/201Tl mismatch). Despite significantly less fibrosis and lower flows, the mismatch segments showed significantly greater. 201Tl activity than the segments with concordantly high tracer retention and flow values. Conversely, at equivalent flow rates, the mismatch regions had less fibrosis than the areas with concordantly depressed 201Tl activity and perfusion. CONCLUSIONS: This super-normal 201Tl retention in hibernating myocardium may indicate a mechanism of cell adaptation to chronic hypoperfusion. PMID- 9747442 TI - Human beta-myosin heavy chain mRNA prevalence is inversely related to the degree of methylation of regulatory elements. AB - OBJECTIVE: Methylation of cytosine in CG dinucleotides within regulatory elements is believed to silence gene expression. These dinucleotides occur in certain important regulatory elements in the promoter region of the human beta-myosin heavy chain (beta-MHC) gene. We therefore investigated whether methylation of these elements correlates with beta-MHC gene transcription in human 'expressing' (right atrial) and 'non-expressing' (peripheral blood leucocytes) cells. METHODS: We employed 2 techniques to assess promoter methylation: (i) analysis of the susceptibility to digestion of a particular CCGG restriction site in the promoter region when genomic DNA is cleaved with the restriction endonucleases MspI (methylation-insensitive) and HpaII (methylation-sensitive), and (ii) the bisulphite-PCR method to examine in detail the methylation patterns of 3 important regulatory elements that contain CG dinucleotides. beta-MHC mRNA expression in right atrium and leucocytes was assessed using reverse transcription-PCR with specific primers that do not detect alpha-MHC cDNA. RESULTS: The digestion pattern observed with MspI or HpaII indicated that the CCGG site was almost completely methylated in leucocytes, but relatively unmethylated in atrial myocardium from the same patients. When methylation was examined with the bisulphite-PCR method we found a reciprocal relationship between the level of beta-MHC mRNA expression in leucocytes and atrial myocardium and the degree of methylation of CG dinucleotides in the 5' regulatory elements of the gene. CONCLUSIONS: Tissue-specific methylation of the human beta-MHC gene promoter may play a role in determining the pattern of expression of this gene. Furthermore, alteration of the level of methylation may underlie the changes in transcription of this gene that occur, for example, when atrial or ventricular myocardium hypertrophies. PMID- 9747443 TI - Pharmacologic activation of the human coronary microcirculation in vitro: endothelium-dependent dilation and differential responses to acetylcholine. AB - OBJECTIVES: In vivo studies of the human coronary resistance circulation cannot control for indirect effects of myocardial metabolism, compression, and neurohumoral influences. This study directly examined the vasodilator responses of the human coronary microcirculation to both receptor-dependent and independent agonists. METHODS: Atrial arterioles were dissected from human right atrial appendage (103 +/- 2 microns diameter, n = 185 vessels from 145 patients) obtained at the time of cardiopulmonary bypass and left ventricular vessels from explanted human hearts (148 +/- 10 microns diameter, n = 57 vessels from 18 patients). After dissection, vessels were mounted onto pipettes in Kreb's buffer under conditions of zero flow and at a constant distending pressure of 60 mmHg. Drugs were applied extraluminally and steady state changes in diameter measured with videomicroscopy. RESULTS: After contraction by endothelin or spontaneous tone, increasing concentrations of adenosine diphosphate (ADP) produced a similar dose-dependent dilation in vessels from atria (maximum 89 +/- 4%, n = 76) and ventricles (maximum 74 +/- 9%, n = 10). The dilation to ADP was abolished by mechanical removal of the endothelium. Similar dilator responses were found to bradykinin, substance P, arachidonic acid, and the calcium ionophore A23187 in both atria and ventricle. In contrast, acetylcholine (ACh) constricted all atrial vessels (-58 +/- 3%, n = 63) regardless of patient age or underlying disease. This constriction was attenuated by denudation, but not affected by inhibition of nitric oxide synthase or cyclo-oxygenase. Microvessels isolated from human ventricle exhibited a heterogeneous response to ACh with dilation being the predominant response. CONCLUSIONS: We conclude that isolated human coronary arterioles demonstrate endothelium-dependent dilation. However, the response to acetylcholine is unique with vasoconstriction in atrial vessels and dilation in ventricular arterioles. PMID- 9747444 TI - Local and systemic delivery of low molecular weight heparin stimulates the reendothelialization after balloon angioplasty. AB - OBJECTIVE: Recent investigations revealed the importance of endothelial cell integrity and function in the pathogenesis of restenosis after angioplasty. Agents which stimulate reendothelialization may prevent restenosis after interventional procedures. The results of in vitro studies suggested that heparin and low molecular weight heparin administration may enhance the recovery of the endothelium. In this study the extent of endothelial denudation and the occurrence and time course of endothelial regeneration after experimental balloon angioplasty followed by subcutaneous or local delivery of low molecular weight heparin was investigated. METHODS: A total of 102 rabbits were included in the study. An atheromatous plaque was induced by electrical stimulation in the right carotid artery of the animals. All animals underwent balloon angioplasty. Thirty two rabbits received no further medical treatment. Twenty-five rabbits received subcutaneous low molecular weight heparin reviparin (400 anti-Xa-units/day) during the following 7 days. In 25 animals the dilated arterial segments were treated locally with reviparin (1500 anti-Xa-units/4 ml, 2 atm) using a porous balloon (2.5 mm, 35 holes, diameter 75 microns). Twenty animals served as control group without intervention. The vessels were excised 3, 7, 14, 28 and 56 days following intervention. Sections were stained with an antibody against von Willebrand factor and PECAM 1 to confirm the endothelial origin of the lining cells. After bromodeoxyuridine labeling, the extent of proliferation was determined by using a monoclonal antibody. In addition, morphometric analysis of the intimal and medial area was performed. RESULTS: Three days after balloon angioplasty histomorphological analysis showed a reduction of about 60% of the preinterventional endothelial cell number in all three groups. Already one week after intervention there was a significantly higher number of endothelial cells in both groups of low molecular weight heparin treated animals compared to the untreated group (s.c. group 144 +/- 33, local group 142 +/- 32 versus untreated 79 +/- 17 endothelial cells, p < or = 0.05). This significant difference was maintained during the following four weeks and demonstrated a 2-fold increase in endothelial proliferation in the heparin treated animals compared with the untreated group. In addition, immunohistological examination showed a significant decrease in smooth muscle cell proliferation in the s.c. and local reviparin treated animals and a subsequent reduction of intimal thickening. CONCLUSION: Local delivery of low molecular weight heparin promotes reendothelialization and contributes to the inhibition of smooth muscle cell proliferation. PMID- 9747445 TI - In vivo cardiovascular reactivity and baroreflex activity in diabetic rats. AB - OBJECTIVES: Abnormalities of the cardiovascular system, e.g. impaired vasoreactivity and changes in baroreflex control of heart rate, are known to occur in experimental diabetes. It is not clear whether these cardiovascular dysfunctions are direct consequences of cardiovascular deficits and/or have autonomic neuropathy as a cause. METHODS: To differentiate between cardiovascular deficits or neuronal impairment as a cause for these cardiovascular dysfunctions, we tested the effects of the ACTH4-9 analogue, Org 2766, a neurotrophic compound without cardiovascular effects, on arterial pressure, heart rate and baroreflex control of heart rate. At 15 weeks, rats were made diabetic by injection of streptozotocin, and from 0-6, 6-12 or 12-18 weeks thereafter 3 groups of rats were treated with Org 2766. These effects were evaluated during phenylephrine induced increases, and sodium nitroprusside-induced decreases, in blood pressure, in rats that had been diabetic for various periods (2-42 weeks). RESULTS: Throughout, both depressor response and maximal vasodilator activity in response to sodium nitroprusside were significantly (P < 0.05) reduced as compared to those of the non-diabetic controls. The pressor response of the diabetic rats to phenylephrine was only significantly (P < 0.05) reduced at 4, 6 and 12 weeks, and at 18 weeks, the diabetic rats were either hypo- or normoresponsive; Org 2766 did not restore the disturbed pressor response. From weeks 4 to 42 both maximal decrease in heart rate and sensitivity of baroreflex-mediated bradycardia in the diabetic rats were significantly less (P < 0.05) than those in the non-diabetic controls. Org 2766 restored the diminished baroreflex-mediated bradycardia of diabetic rats to non-diabetic control levels at 6 weeks, had an ameliorating effect at 12 weeks and no effect at 18 weeks. CONCLUSIONS: Time-dependent decreases in baroreflex sensitivity in diabetic rats was demonstrated and a much less steep decline of baroreflex sensitivity occurred in non-diabetic control rats. The ACTH4-9 analogue, Org 2766, when given immediately upon the induction of diabetes seem to delay the development of autonomic neuropathy, which suggests that cardiovascular factors appear to be of minor importance. PMID- 9747446 TI - Morphological and functional changes in coronary vessel evoked by repeated endothelial injury in pigs. AB - OBJECTIVE: We examined the morphological changes induced by repeated endothelial denudation in coronary artery (CA), as well as functional changes in the endothelium-dependent and smooth muscle responses to various vasoactive agents during the process of intimal thickening. METHODS: We observed vascular responses in denuded and non-denuded portions of pig CA while being fed a normal diet (n = 11, N group) or 2% cholesterol diet (n = 25, C group) to intracoronary acetylcholine (ACh), 5-hydroxytryptamine (5-HT), substance P (SP), and isosorbide dinitrate (ISDN) with and without the nitric oxide synthesis inhibitor N omega nitro-L-arginine methyl ester (L-NAME, 10 mg/kg i.v.) over a period of 8 weeks. Balloon endothelial denudation of the left anterior descending CA was carried out every 2 weeks. RESULTS: In N group, maximum vasoconstriction was obtained with ACh 2 weeks after the first denudation [26 +/- 5% vs. 1 +/- 1% pre-denudation, p < 0.05]. L-NAME did not affect ACh-induced CA diameter changes. Thereafter, the response to ACh was attenuated by repeated denudation in N groups. However, the degree of 5-HT-induced CA narrowing at the denuded portion increased from 7 +/- 4% (0 week) to 88 +/- 8% (8 weeks) (p < 0.05). The changes resulted in severe myocardial ischaemia, and suggested that endothelium-dependent vasodilation was progressively attenuated while hyperreactivity of vascular smooth muscle simultaneously increased. Vasodilation induced by SP was attenuated somewhat, but ISDN-induced vasodilation was preserved. Although mild hypercholesterolaemia was induced in C group, the vascular responses to these vasoactive agents did not differ from those of N group. CONCLUSIONS: Repeated CA endothelial injury and regeneration induce the change of morphology and vascular reactivity in the denuded portion regardless of atherogenic diet. This study strongly suggests that intimal thickening caused by repeated endothelial injury and regeneration induces specific vascular responses to vasoactive agents. Moreover, it is also suggested that during the progression of intimal thickening, increased vascular smooth muscle contraction and decreased endothelium-dependent dilation appear in a stimulus-dependent manner, often leading to severe coronary vasoconstriction accompanied with definitive ECG ST change. PMID- 9747447 TI - Permissive effect of nitric oxide in arachidonic acid induced dilation in isolated rat arterioles. AB - OBJECTIVE: Prostaglandins and nitric oxide play an important role in the regulation of arteriolar tone. L-Arginine analogues inhibit nitric oxide formation, but may also inhibit arachidonic-acid induced dilation. Nitric oxide was found to stimulate cyclooxygenase activity in cultured endothelial cells. Therefore, we hypothesized that the non-specific inhibition of prostaglandin related dilation by L-arginine analogues is a consequence of the absence of nitric oxide. METHODS: To test this hypothesis, arteriolar segments from rat cremaster muscle were studied in a pressure myograph at 75 mmHg. Segments developed spontaneous tone, the diameter reduced from 179 +/- 3 to 98 +/- 3 microns (n = 41). In this condition, responses to exogenous arachidonic acid (1 microM) were recorded and compared with responses after addition of L-NNA, and addition of either SNAP, nitroprusside or 8-Br-cGMP in the presence of L-NNA. RESULTS: Inhibition of basal nitric oxide production with L-NNA (0.1 mM) reduced arachidonic acid-induced dilation (from 52 +/- 9 to 31 +/- 6 microns). In the presence of L-NNA, responses to arachidonic acid were augmented when exogenous nitric oxide was also present (SNAP, 31 +/- 6 microns vs. 75 +/- 5 microns; nitroprusside, 31 +/- 8 microns vs. 42 +/- 7 microns). Responses were not augmented with the second messenger of nitric oxide-mediated dilation 8-Br-cGMP (37 +/- 9 microns vs. 32 +/- 9 microns). CONCLUSIONS: These results indicate that nitric oxide directly increases arachidonic acid-induced dilation. Thus, the non specific effect of L-arginine analogues can be explained by a permissive effect of nitric oxide on endothelial arachidonic acid metabolism. PMID- 9747448 TI - Characterization of the hyperpolarization-activated inward current in isolated human atrial myocytes. AB - OBJECTIVE: The hyperpolarization-activated inward current (I(f)) has been discussed to contribute to arrhythmias in rat hypertrophied and human failing ventricular myocardium. Cat atrial myocytes were found to exhibit variable size of I(f). In the present study, we evaluate characteristics of I(f) in human atrial myocardium and investigate if human atria might exhibit any electrophysiological heterogeneity in the diastolic voltage range. METHODS AND RESULTS: The whole-cell patch-clamp technique was used to record I(f) in isolated myocytes from 96 human right atrial appendages. I(f) was observed in 95% (ruptured-patch; 141/146) to 100% (perforated-patch; 18/18) of myocytes showing typical current properties, i.e. time- and voltage-dependence, block by [Cs+]o, permeability for K+, Na+ and Li+, and current increase with raising [K+]o. Using the perforated-patch technique Boltzmann distributions yielded an activation threshold of -60 to -70 mV and half maximal activation at -89.3 +/- 0.7 mV (n = 18). Isoproterenol (10(-6) mol/l) shifted I(f) activation by +7 mV (7/7) using the perforated-patch technique, but only inconsistently shifted I(f) activation using the ruptured-patch method (6/21). Based on the relative current size of I(f) and IK1 three cell types could be distinguished (n = 26). In myocytes with a prominent I(f), I(f) density was found to be larger (in [K+]o 25 mmol/l at -80 mV: -0.78 +/- 0.23 pApF-1; n = 7) than in cells with predominant IK1 (in [K+]o 25 mmol/l at -80 mV: -0.02 +/- 0.01 pApF-1; n = 4) (P < 0.05). CONCLUSIONS: I(f) is present in most human atrial myocytes. Many current properties are similar to those described for I(f) in mammalian pacemaker cells. The relative current size of I(f) and IK1 were found to be variable in different myocytes. Whether I(f) may favor spontaneous diastolic depolarization in individual human atrial myocytes exhibiting predominantly I(f) in vivo remains to be defined, as current size is very small under physiological [K+]o. PMID- 9747449 TI - Molecular cloning of genes differentially regulated by TNF-alpha in bovine aortic endothelial cells, fibroblasts and smooth muscle cells. AB - OBJECTIVE: Tumor necrosis factor-alpha (TNF-alpha) is a pleiotropic-cytokine binding to and thereby stimulating vascular cells. TNF-alpha mediated intermediate stimulation of vascular cells is believed to play a pivotal role in the development of arteriosclerosis. While extensive information has recently become available on gene induction by TNF-alpha, less is known about gene suppression by TNF-alpha in vascular cells. Endothelial cells are the first cell layer within the vessel wall interacting with circulating, cytokine releasing cells. Therefore, they were selected as target for these study. METHODS: A differential screening approach has been used to isolate cDNAs whose abundance was suppressed by incubating bovine aortic endothelial cells (BAEC) for 6 h with 1 nM TNF-alpha. The gene expression of 6 isolated cDNAs after TNF-alpha was investigated by dot blots and nuclear run-on analysis in BAEC. The investigated genes were partially or completely sequenced. Differential expression after TNF alpha stimulation of BAEC, bovine fibroblasts and vascular smooth muscle cells (SMC) was studied by Northern blots. RNA transcripts of the clone C7 in aortic aneurysms were examined by in situ hybridization. RESULTS: 49 independent cDNAs were isolated by the differential screening approach and 6 clones were further analyzed. These genes were downregulated in a time and dose dependent manner in BAEC. Sequence analysis revealed that 3 cDNAs encoded previously unidentified genes (C1, C5, C7), while 3 encoded known genes: connective tissue growth factor (CTGF; A1), fibronectin (A8) and the mitochondrial genome (B1). A1 and B1 were suppressed in BAEC, fibroblasts and SMC, whereas A8, C1, C5 and C7 were not uniformly downregulated in the investigated cells. C7 RNA transcripts were exclusively induced in the endothelium of an uninflamed aortic aneurysm. The transcripts were undetectable in an inflamed aortic aneurysm and control vessels. CONCLUSIONS: Gene suppression is a prominent feature of the intermediate effect of TNF-alpha on endothelial cells. Differences in the expression of the tested genes in endothelial cells, fibroblasts and vascular smooth muscle cells open possibilities for the study of cellular interactions in the vascular wall in disease situations with high local TNF-alpha concentrations. PMID- 9747450 TI - Induction of nitric oxide synthase in human vascular smooth muscle: interactions between proinflammatory cytokines. AB - OBJECTIVE: We have attempted to demonstrate the induction of inducible nitric oxide synthase in human vascular tissue and define the capacity of different cytokines to induce this enzyme. METHODS: Segments of human arteries were stimulated with lipopolysaccharide (10 micrograms/ml), interleukin-1 beta (5 U/ml), tumor necrosis factor-alpha (10 U/ml), and interferon-gamma (200 U/ml). Cytokines were either used alone or in certain combinations, as well as in the presence of L-NG-monomethyl-arginine (100 mumol/l) or cycloheximide (1 mumol/l). Induction was assessed by measurement of mRNA expression, immunocytochemical localisation of the expressed protein, nitric oxide synthase activity and levels of nitrite, a product of nitric oxide formation. RESULTS: PCR analysis showed the presence of mRNA for iNOS in stimulated samples which could be inhibited by cycloheximide. There was positive staining with an antibody against human iNOS in the media of stimulated vessel segments. Stimulated segments were also shown to contain Ca(2+)-independent nitric oxide synthase activity. The cytokines and lipopolysaccharide together gave a significant rise in levels of nitrite in the medium after 36 and 48 h, which was inhibited by L-NG-monomethyl-arginine and cycloheximide. Only interferon-gamma incubated alone was capable of increasing nitrite levels. This effect was enhanced by co-incubation with either interleukin 1 beta, tumor necrosis factor-alpha or lipopolysaccharide. CONCLUSION: We have shown that increased production of nitrite by human vascular tissue in response to cytokines is associated with induction of iNOS as shown at the molecular and protein levels, and further supported by the presence of increased Ca(2+) independent nitric oxide synthase activity following cytokine stimulation. PMID- 9747451 TI - Adenoviral vectors: not to be sneezed at. PMID- 9747452 TI - Cell death and cancer: replacement of apoptotic genes and inactivation of death suppressor genes in therapy. AB - This review provides a critical evaluation of the increasing use of gene therapy in the treatment of malignancies to induce active cell death (ACD, apoptosis). This approach is consistent with the notion that cancer is an anomalous accumulation of cells largely resulting from diminished cell death. The review details the main genes potentially useful for therapy. Among these, p53 has received the majority of the investigators' attention and provided encouraging results. Even greater hope is offered by newly tried direct inducers of apoptosis, such as bax, bclXs and caspases. Another fruitful direction is the association of apoptosis-inducing gene transfer with radio- and chemotherapy, which are also inducers of ACD. There is a delicate balance between cell gain through mitosis and cell loss in neoplasia because spontaneous apoptosis is widely present in tumors. In fact, the tumor environment favors bystander cell killing which appears to be a fundamental mechanism insofar as the rate of observed cell mortality cannot be accounted for by the known methods of gene transduction with efficiencies far below 100%. We conclude that apoptosis offers a mainstream approach for cancer gene therapy since ACD is highly inducible and only limited gains in malignant cell apoptosis may displace tumors from growth to regression. PMID- 9747453 TI - Poly (lactic-glycolic) acid copolymer encapsulation of recombinant adenovirus reduces immunogenicity in vivo. AB - Adenovirus-mediated gene transfer has application to the treatment of diseases of the central nervous system. We demonstrate that a limitation to its use in vivo is an inability to redose to the brain. We show that one factor inhibiting re dosing is the development of neutralizing anti-adenoviral antibodies. Encapsulation of recombinant adenovirus vectors in poly(lactic/glycolic acid) (PLGA) copolymer enables infection in vitro, in the presence of neutralizing antibodies and results in the release of viable virus for over 100 h. Importantly, encapsulated adenovirus also shows diminished immunogenicity in vivo. Mice immunized with encapsulated recombinant adenoviral vectors show a greater than 45-fold reduction in anti-adenovirus titers relative to non encapsulated vectors. An extended release formulation of adenovirus that reduces viral immunogenicity and sequesters the viral particle form antibody exposure may improve in vivo efficacy. PMID- 9747454 TI - Adenovirus-mediated wild-type p53 overexpression inhibits endothelial cell differentiation in vitro and angiogenesis in vivo. AB - Gene therapy with the tumor suppressor gene p53 induces cancer cell apoptosis in vitro and in vivo and inhibits tumor growth in nude mice. We hypothesized that, in addition to cancer cell apoptosis, a replication-deficient adenovirus vector which carries the cDNA for human wild-type p53 (AdCMV.p53) may also modulate endothelial cell function and inhibit angiogenesis. Human umbilical vein endothelial cells (HUVEC) were infected at different multiplicities of infection (MOI) with either AdCMV.p53, the control vector AdCMV.null or were not infected. Western blot analysis showed p53 overexpression up to 7 days after infection with AdCMV.p53. HUVEC proliferation was either not affected (20 and 50 MOI) or inhibited to comparable levels (100 MOI; P < 0.05) in AdCMV.p53- and AdCMV.null infected versus uninfected cells. HUVEC differentiation into capillary-like structures on reconstituted basement membrane proteins (Matrigel) was assessed 48 h after infection (100 MOI). After 18 h on Matrigel the capillary-like network formed by AdCMV.p53-infected HUVEC was less extensive than that formed by both AdCMV.null-infected and uninfected control cells (P < 0.05 versus either control). In contrast, conditioned medium from AdCMV.p53-infected HUVEC did not modulate endothelial cell differentiation on Matrigel. The effect of AdCMV.p53 on angiogenesis in vivo was assessed by injecting this vector subcutaneously in mice; 3 days later Matrigel containing basic fibroblast growth factor (bFGF) was injected at the same site. In other experiments AdCMV.p53 was injected simultaneously with an Ad vector coding for vascular endothelial growth factor (AdCMV.VEGF165) into the rat perirenal fat tissue. AdCMV.p53 significantly inhibited neovascularization induced by bFGF within the Matrigel plugs (P < 0.05) or by AdCMV.VEGF165 in the fat tissue (P < 0.05). Thus, the anti-angiogenic effect of Ad-mediated wild-type p53 overexpression may contribute to the ability of this viral vector to inhibit tumor growth. PMID- 9747455 TI - A novel method for the direct quantification of gene transfer into cells using PCR in situ. AB - There are several limitations to current methods for the detection of target genes following gene transfer. We report a novel PCR in situ procedure which overcomes many of these and permits the direct quantification of gene transfer in individual cells. PCR amplification of a proviral specific nucleotide sequence in target cells was followed by in situ hybridization using fluorescent probes complementary to different regions of the amplicon. Many of the problems previously encountered using in situ PCR, particularly the generation of false positive results and extracellular leakage of PCR products, were overcome by modifications of existing protocols. Positive cells were readily identified by fluorescence microscopy and a high sensitivity, specificity and correlation coefficient were demonstrated in mixing experiments using varying proportions of known provirus positive and negative cells. The method was applied successfully to identify low numbers of gene-modified hematopoietic cells in clinical specimens in a trial of retrovirus-mediated gene transfer into blood forming stem cells. This approach is simple and reliable, has the potential for use in a variety of gene therapy applications and may become the method of choice for the assessment of gene transfer efficacy. PMID- 9747456 TI - Inhibition of intimal hyperplasia after vein grafting by in vivo transfer of human senescent cell-derived inhibitor-1 gene. AB - The senescent cell-derived inhibitor (sdi)-1 (p21) protein has been identified as a downstream mediator of the tumor suppressor p53 in the cell cycle regulation. In this study, we focused on the function of sdi-1 gene in inhibiting vascular smooth muscle cell (VSMC) proliferation after vein grafting in a rabbit model. To test the hypothesis, we transfected human sdi-1 gene by an intra-operative approach. Accompanied by markedly increased sdi-1 protein, the significant increase in PCNA-stained VSMCs in vein grafts was inhibited by transfection of sdi-1 gene. Moreover, at 2 weeks after transfection, transfer of sdi-1 gene resulted in a significant inhibition in neointimal formation, compared with control vector. Of importance, immunohistological studies determining the expression pattern of myosin heavy isoforms, adult type specific SM2 and embryonic specific SMemb/NMHC-B, demonstrated expression of the adult phenotype of VSMCs in the neointima of sdi-1 gene-transfected vein grafts at 2 weeks after the operation, while the neointima was predominantly composed of embryonic phenotype of VSMCs in the control grafts. Overall, these results demonstrate that a single intraluminal incubation of human sdi-1 gene can result in a significant inhibition of neointimal formation after vein grafting, associated with phenotypic change of VSMCs from neonatal to adult type in a rabbit model. Inhibition of hyperplasia in a graft model by transfection of sdi-1 gene may be due to the change in VSMC phenotype from neonatal to adult, in addition to the inhibition of VSMC growth. PMID- 9747457 TI - Recombinant adenoviral delivery for in vivo expression of scFv antibody fusion proteins. AB - Antibodies and their recombinant fragments have enormous potential for therapy of malignant and other diseases, but there can be problems associated with their production and purification in the quantities required for therapeutic use. We investigated the use of gene therapy for the production of such recombinant antibody fragments in vivo. We generated two recombinant adenoviruses expressing the single chain Fvs (scFvs) fused to murine GM-CSF (mGM-CSF). The scFvs used are MFE-23 which binds to a human tumour-associated marker carcino-embryonic antigen (CEA) and B1.8 which binds the hapten 4-hydroxy-3-nitro-5-iodo-phenylacetyl (NIP). Using scFvs to target GM-CSF to tumour cells should reduce the systemic toxicity of GM-CSF but retain its ability as a cytokine to induce systemic immune responses to tumour targets. Cell lines infected with the recombinant adenoviruses in vitro express and secrete high levels of the scFv.mGM-CSF fusion proteins. The scFv retains specificity while the mGM-CSF portion is fully bioactive and there is no detectable degradation of the fusion product. We also demonstrated effective in vivo expression of the scFv.mGM-CSF proteins. C57BI/6 mice injected intravenously with the adenovirus encoding the MFE-23.mGM-CSF fusion produce high levels of the fusion protein by 2 days after infection. The scFv.mGM-CSF protein can be detected in the serum, at biologically active levels, for at least 20 days and the level of protein produced is related to the amount of adenovirus injected. This approach has the potential to streamline the testing of the many therapeutic strategies based on recombinant scFvs and we are currently testing these constructs in an animal model for antitumour activity. PMID- 9747458 TI - IL-1/IL-3 gene therapy of non-small cell lung cancer (NSCLC) in rats using 'cracked' adenoproducer cells. AB - Cytokine gene therapy was studied in established L42 tumours in syngeneic rats. L42 is a transplantable non-immunogenic non-small cell lung cancer (NSCLC). Genes coding for human interleukin-1 alpha and for rat interleukin-3 beta were transferred by injecting producer cells of recombinant adenovirus vectors into the tumour in attempts to achieve high concentrations of the cytokines inside the tumor without systemic toxicity. Limited tumour growth delay was obtained with viable producer cells. For logistic reasons stocks of pooled frozen producer cells allowed intensive treatment of groups of tumour bearing rats. The cells were lysed by thawing before administration. Ten daily injections of such 'cracked' producer cells induced reproducible tumour responses. These were due to local release of cytokines, not to systemic effects. Growth retardation also occurred in contralateral tumours which were not injected. When rats carrying established tumours were vaccinated with lysates of tumours collected during treatment with 'cracked' producer cells, significant tumour growth retardation was obtained. We speculate that both cytokines, if produced at sufficiently high concentrations in tumours, induce inflammation which in turn initiates an immune response against tumours growing at a distant site. These findings seem to justify further exploration of IL-1 and IL-3 gene transfer for the treatment of cancers. PMID- 9747459 TI - Superiority of the ear pinna over muscle tissue as site for DNA vaccination. AB - Three different vaccination sites were compared for efficiency of immunization with naked DNA. Using the bacterial lacZ gene as a model, all three sites of the mouse (skeletal muscle, dermis of abdominal skin or of the ear pinna) could express the gene product beta-gal but varied in expression time with muscle tissue showing the longest expression. Expression time, however, did not correlate with immune response intensity. The ear pinna was by far the most effective and muscle the least effective priming site for specific humoral and cytotoxic T cell-mediated immune responses. Following intra-pinna DNA inoculation, beta-gal expressing cells were detectable around the injection site and in the major draining lymph node. Efficiency of immunization was also dependent on the promoter and expression vector used. The cytomegalus virus promoter driven pCMV beta vector was superior to the Moloney murine leukemia virus LTR driven BAG vector. LacZ DNA immunization was also compared with cell based vaccination with lacZ-transfected tumor cells, in which case again the pinna was the best site for inducing strong immune responses. Tumor-specific T cell responses could also be well induced in the pinna, leading to cytotoxic T lymphocyte induction and protective antitumor immunity. Thus, the pinna was found to be a privileged site for induction of antitumor responses and for genetic immunization, an important finding of immediate practical and potential future clinical implications. PMID- 9747460 TI - Imaging and tissue biodistribution of 99mTc-labeled adenovirus knob (serotype 5). AB - Hepatic sequestration of systemically administered adenoviral vectors reduces the number of viral particles available for delivery to other tissues. The biological basis of this phenomenon was investigated using a new in vivo technique which permitted imaging in real time. Recombinant adenovirus serotype 5 knob (Ad5K) was radiolabeled with the gamma-emitter 99mTc (half-life = 6 h). Scatchard analysis of the 99mTc-Ad5K showed specific, high-affinity binding to U293 cells (Kd = 1.4 +/- 0.5 nM), demonstrating that the radiolabeling process had no effect on receptor binding. In vivo dynamic imaging with an Anger gamma camera revealed that the liver binding followed an exponential rise to maximum, with a measured 100% extraction efficiency. Initially, the liver binding capacity was 3.1 +/- 0.4 micrograms Ad5K, equivalent to approximately 17,000 Ad5K molecules per liver cell. Liver binding was blocked by preincubation of Ad5K with neutralizing anti Ad5K antibody; a 50% reduction in liver uptake was demonstrated by imaging. Unlabeled Ad5K was more effective in blocking liver uptake of 99mTc-Ad5K, whereas irrelevant unlabeled Ad3K had no effect. Imaging data for the liver uptake studies were in agreement with biodistribution determined by removing and measuring tissues. These data demonstrated that in vivo imaging is a sensitive tool for measuring changes to liver tropism. Similar imaging techniques can be applied to adenovirus vectors to measure specific targeting for gene therapy. PMID- 9747462 TI - In vivo expression of therapeutic human genes for dopamine production in the caudates of MPTP-treated monkeys using an AAV vector. AB - An adeno-associated virus (AAV) vector, expressing genes for human tyrosine hydroxylase (TH) and aromatic amino acid decarboxylase (AADC), demonstrated significantly increased production of dopamine in 293 (human embryonic kidney) cells. This bicistronic vector was used to transduce striatal cells of six asymptomatic but dopamine-depleted monkeys which had been treated with the neurotoxin MPTP. Striatal cells were immunoreactive for the vector-encoded TH after stereotactic injection for periods up to 134 days, with biochemical effects consistent with dopamine biosynthetic enzyme expression. A subsequent experiment was carried out in six more severely depleted and parkinsonian monkeys. Several TH/aadc-treated monkeys showed elevated levels of dopamine near injection tracts after 2.5 months. Two monkeys that received a beta-galactosidase expressing vector showed no change in striatal dopamine. Behavioral changes could not be statistically related to the vector treatment groups. Toxicity was limited to transient fever in several animals and severe hyperactivity in one animal in the first days after injection with no associated histological evidence of inflammation. This study shows the successful transfection of primate neurons over a period up to 2.5 months with suggestive evidence of biochemical phenotypic effects and without significant toxicity. While supporting the idea of an in vivo gene therapy for Parkinson's disease, more consistent and longer lasting biochemical and behavioral effects will be necessary to establish the feasibility of this appraoch in a primate model of parkinsonism. PMID- 9747461 TI - Pre-existing herpes simplex virus 1 (HSV-1) immunity decreases, but does not abolish, gene transfer to experimental brain tumors by a HSV-1 vector. AB - The influence of pre-existing anti-herpes simplex type 1 (HSV-1) immunity on HSV 1 vector-mediated gene transfer to glioma cells was analyzed in this gene marking study using intracranial D74 gliomas in syngeneic Fischer rats. The HSV-1 mutant virus used, hrR3, is defective in ribonucleotide reductase and bears the marker genes E. coli lacZ and HSV-1 thymidine kinase (HSVtk). Initial marker gene expression in tumors 12 h after direct virus injection was reduced in immunized animals to about 15% of that in nonimmunized animals. Marker gene expression in both sets stayed at initial levels for 2 days after intratumoral injection and declined markedly on day 5. Inflammatory infiltrates in the tumor were more prominent in HSV-1-immunized, as compared with nonimmunized animals, at 12 and 24 h, but appeared similar at 2-5 days after injection. By day 10, the immune reaction had subsided in immunized animals and macrophages remained only in nonimmunized animals. In conclusion, gene transfer to brain tumors using a HSV-1 vector was greatly reduced, but not completely abolished, under pre-immunization conditions. Pre-existing antibodies to HSV-1 may also serve a positive role in providing an increased margin of safety in intracranial application of HSV-1 vectors by limiting spread of the virus within the brain and to other tissues. PMID- 9747463 TI - Systemic long-term delivery of antibodies in immunocompetent animals using cellulose sulphate capsules containing antibody-producing cells. AB - Implantation of capsules containing antibody-producing cells into patients would potentially permit systemic long-term delivery of antibodies and might, thus, be useful in the development of surveillance treatments for cancers and severe viral diseases. We show that cellulose sulphate (CS) capsules containing hybridoma cells, when implanted subcutaneously or in the intraperitoneal cavity, can be used for delivering monoclonal antibodies into the blood-stream of immunocompetent mice for at least several months. In contrast to capsules implanted into the intraperitoneal cavity, which remain mobile and nonvascularized, capsules implanted under the skin form neo-organs which become vascularized within days. This may explain the higher blood concentration of the antibody we have observed in the latter case. Importantly, neither an isolating fibrosis nor an obvious inflammatory response was detected at the capsule implantation sites during observation periods as long as 10 months. Finally, no anti-idiotypic immune response against the ectopically delivered antibody was shown to occur. This rules out any potent adjuvant effect of the cellulose sulphate matrix that might have stimulated a neutralizing humoral response. Taken together, our data indicate that encapsulation of antibody-producing cells into CS might be used in antibody-based gene/cell therapy approaches. PMID- 9747464 TI - Chemoprotective gene transfer I: transduction of human haemopoietic progenitors with O6-benzylguanine-resistant O6-alkylguanine-DNA alkyltransferase attenuates the toxic effects of O6-alkylating agents in vitro. AB - Retroviral transduction was used to introduce cDNAs encoding two mutants of human O6-alkylguanine-DNA alkyltransferase (hAT), one of which (hATPA) is 16 times more resistant to O6-benzylguanine (O6-beG), and the other (hATPA/GA) which is almost totally refractory to inactivation relative to the wild-type protein, into K562 human erythroleukaemic cells. A colony-forming assay was used to demonstrate significant protection (P < 0.001) against mitozolomide or temozolomide toxicity in K562 clones expressing either hAT mutant, as determined from an in vitro assay of activity. However, protection against these agents was reduced in hATPA expressing cells in the presence of 1 microM O6-beG and was lost in the presence of 20 microM O6-beG while cells expressing hATPA/GA retained protection even in the presence of 20 microM O6-beG (P < 0.001). Using primary human cord blood derived CD34+ haemopoietic cells in which PCR analysis indicated that up to 70% of progenitors were transduced with retroviral constructs harbouring hATPA/GA, we observed significant protection of the granulocyte-macrophage colony-forming cells against mitozolomide (P < 0.05) and temozolomide (P < 0.001) induced toxicity in the presence of O6-beG. These findings indicate that retrovirus mediated expression of hATPA/GA in primitive primary human haemopoietic cells is possible and does provide O6-beG-resistant protection for these cells. Using this strategy in patients may simultaneously permit attenuated myelosuppression and increased sensitivity of tumour cells to the effects of O6-alkylating agent chemotherapy. These data, taken together with the study reported by Chinnasamy et al in the accompanying article in this issue showing reduced toxicity and clastogenicity in murine haemopoietic progenitors, make a compelling case to test this strategy clinically. PMID- 9747465 TI - Chemoprotective gene transfer II: multilineage in vivo protection of haemopoiesis against the effects of an antitumour agent by expression of a mutant human O6 alkylguanine-DNA alkyltransferase. AB - Murine bone marrow cells were transduced ex vivo with a retrovirus encoding an O6 benzylguanine (O6-beG) insensitive, double mutant form of the human DNA repair protein O6-alkylguanine-DNA alkyltransferase (hATPA/GA). In animals reconstituted with the transduced bone marrow, about 50% of cells in the multipotent spleen colony-forming cells (CFU-S) and lineage restricted granulocyte-macrophage (GM CFC) haemopoietic progenitor populations were found to be carrying the transgene and this correlated with the frequency of bone marrow cells and spleen colonies which stained positive for hATPA/GA by immunocyto-chemistry. Expression of hATPA/GA was associated with significant in vivo protection of both CFU-S (P = 0.001) and GM-CFC (P < 0.024) against the toxicity of the antitumour methylating agent, temozolomide, given in combination with O6-beG. Expression of hATPA/GA also led to a reduction in the frequency of combined O6-beG/temozolomide-induced micronuclei seen in polychromatic erythrocytes (P < 0.003). This study is the first to demonstrate in vivo protection of multipotent haemopoietic progenitors against the toxic and clastogenic effects of an O6-alkylating agent in the presence of O6-beG. It also represents the first report of reduced clastogenesis as a consequence of expression of an O6-beG-resistant ATase. In the accompanying article we report hATPA/GA-mediated resistance of human CD34+ haemopoietic progenitors to combined O6-beG/O6-alkylating agent toxicity. Together these two reports suggest that a gene therapy strategy whereby protection of normal haemopoietic tissue may be combined with O6-beG-mediated tumour sensitisation may be efficacious in achieving an increase in therapeutic index. PMID- 9747467 TI - Influence of the DNA complexation medium on the transfection efficiency of lipospermine/DNA particles. AB - Dioctadecylamidoglycylspermine (DOGS, Transfectam) is a cationic lipid able to interact with DNA to form complexes that mediate efficient gene transfer into various eukaryotic cells. The state of condensation of the plasmid changes with the medium composition. We therefore investigated to what extent the DNA condensation buffer influences the transfection efficiency of Transfectam/DNA particles. Our results show that in a variety of cell lines, a greater than 100 fold difference in luciferase gene expression is observed with Transfectam/DNA complexes at a +/- charge ratio of 0.75 depending on the conditions of complex formation. The best transfection conditions consisted of particles formed in RPMI medium, NaHCO3/Na2HPO4 or sodium citrate solutions. Mixing in a 150 mM sodium chloride solution (as recommended) resulted in lower gene expression. When the helper lipid 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) was present in the DNA/cationic lipid formulation, the increase in reporter activity was also observed, although to a lower extent. Thus, choosing the optimal conditions for formulating DNA/lipid complexes considerably reduces the amount of lipid and DNA needed to obtain maximum gene transfer. PMID- 9747466 TI - Targeting of a cholecystokinin-DNA complex to pancreatic cells in vitro and in vivo. AB - The carboxy terminal octapeptide of cholecystokinin (CCK8) is a hormone that binds high affinity receptors in a number of tissues including pancreas and pancreatic tumours. As part of our studies to develop effective gene therapy for the treatment of pancreatic cancers, we have investigated various gene delivery systems that depend on CCK8 receptor targeting. In this paper, we describe the synthesis of a CCK8-DNA complex designed to deliver foreign DNA to cholecystokinin receptor-positive cells. CCK8 was ligated to avidin and then complexed to linearised biotinylated DNA (pSV-CAT). The uptake of 32P-labelled CCK8-DNA complex by rat pancreatic acini was linear with time over 4 h with 65 70% of uptake inhibited by 100 nM CCK8. The complex appeared to be internalised since it could not be removed by acid wash. When administered intra-arterially, the complex was rapidly removed from the circulation with no evidence of targeted delivery to the pancreas. However, following a single intraperitoneal dose, the pancreas accumulated 5-8% of the total administered complex by 24 h. These results suggest that peptide-dependent gene delivery to CCK receptor positive cells in vivo is feasible but, when administered directly into the circulation, diffusional barriers across the endothelium may limit distribution to peripheral tissues. Intraperitoneal administration therefore may be a useful alternative for targeting the pancreas. PMID- 9747468 TI - Treatment of sequelae after facial paralysis: a global approach. PMID- 9747469 TI - The molecular genetics of inherited deafness--current knowledge and recent advances. PMID- 9747470 TI - Level I dissection for laryngeal and hypopharyngeal cancer: is it indicated? AB - Squamous cell carcinoma of the larynx and hypopharynx tends to metastasize frequently to cervical lymph nodes, the location of which depends mainly on the site of the primary lesion. Five anatomical levels of cervical nodes have consequently been defined to standardize the terminology used to describe which lymph node groups are at risk for metastatic spread. Level I includes the submental and submandibular triangles. This review considers the role of these triangles in neck dissection and concludes that, unless there is clear evidence of spread, the inclusion of the level I triangles in the neck dissection is unwarranted since these nodes are not really at risk. There is therefore an important role for selective neck dissection in suitable cases of squamous cell carcinoma of the larynx and hypopharynx. PMID- 9747471 TI - Pre-operative and per-operative factors conditioning long-term facial nerve function in vestibular schwannoma surgery through translabyrinthine approach. AB - Facial nerve function was evaluated in 103 patients, after vestibular schwannoma removal through the translabyrinthine approach. The mean follow-up was 43 months (minimum six months). Grade I facial function was achieved in 100 per cent of stage I schwannomata compared with 36 per cent of stage IV schwannomata. Grade I or II facial function was found in 78 per cent of homogeneous schwannomata, compared with 48 per cent of heterogeneous schwannomata. Facial function was preserved in 89 per cent of cases, if the angle between the internal auditory canal and the schwannoma was > 66 degrees, compared with 54 per cent if the angle was < 66 degrees. There was 82 per cent of normal facial function when the nerve appeared normal after tumour removal, compared with 18 per cent when the nerve was traumatized. When the ratio (stimulation threshold at the internal auditory canal/stimulation threshold at brainstem) was < 2, post-operative facial function was preserved in 87 per cent of cases, compared with 13 per cent when the ratio was > 2. PMID- 9747472 TI - Combined transfacial and neurosurgical approach to malignant tumours of the ethmoid sinus. AB - In order to understand the risks and benefits of a combined transfacial and neurosurgical procedure for neoplasms of the ethmoid sinus, we reviewed all patients who underwent this surgical approach in our department between 1986 and 1994. The study included 41 patients. Pathological diagnoses included adenocarcinoma (31 patients), squamous cell carcinoma (three patients), aesthesioneuroblastoma (three patients), other (four patients). The overall morbidity rate was 39 per cent, and the post-operative mortality rate was 2.5 per cent. Complications were statistically more likely in patients with bone skull base reconstruction. The main carcinologic failures were local recurrences (24 per cent) and metastases (22 per cent). The one-year, three-year and five-year Kaplan Meir survival rates were respectively 84 per cent, 53 per cent and 36 per cent. In conclusion, the mortality and morbidity were acceptable, especially when no bone skull base reconstruction was performed. Better local control justifies a combined procedure with post-operative radiotherapy when tumours involve or reach the skull base. PMID- 9747473 TI - Changes in the male voice at puberty: vocal fold length and its relationship to the fundamental frequency of the voice. AB - Ultrasound measurements of the vocal folds were taken for a number of boys passing through puberty. The boys were grouped according to their pubertal stage as defined by Tanner and there was a gradual increase in the length of the vocal folds as puberty progressed. The fundamental frequency of the boys' speaking voice was recorded via laryngography and a good correlation between the length of the vocal folds and the frequency of the voice was seen. The sudden drop in frequency seen between Tanner stages 3 and 4 did not correlate with similar changes in the length of the vocal folds at this time but stroboscopic findings suggest a change in the structure and mass of the vocal folds at this time of maximum frequency change. PMID- 9747474 TI - The predictive value of p53, MDM-2, cyclin D1 and Ki67 in the progression from low-grade dysplasia towards carcinoma of the larynx. AB - To evaluate the predictive role of the oncogenes p53, MDM-2 and cyclin D1, and the proliferative marker Ki67, in the progression from low-grade dysplasia to carcinoma of the larynx. We studied immunohistochemically a series of 32 low grade pre-neoplastic laryngeal lesions, 10 of which progressed to invasive carcinoma. Immunoreactivity in more than 10 per cent of the dysplastic cells was detected in five cases immunostained with anti-p53 (approximately 15 per cent), in two with anti-MDM-2 (approximately six per cent), and 11 with anti-Ki67 antibodies (approximately 34 per cent), whereas none of the cases showed cyclin D1 overexpression. No significant association was found between p53 and MDM-2 immunoreactivity and the evolution to carcinoma; on the contrary, Ki67 expression was detectable in all but one of the 10 cases developing an infiltrative tumour (90 per cent), and in two of the 22 cases that did not progress (approximately nine per cent) (p = 0.01). These findings indicate that immunohistochemical assessment of the proliferative index in bioptic samples of dysplastic laryngeal mucosa may be useful in selecting patients who should undergo a more specific follow-up evaluation. PMID- 9747475 TI - A qualitative assessment of randomized controlled trials in otolaryngology. AB - In 1996 the CONSORT statement made recommendations on the strict reporting of randomized controlled trials (RCT). This will facilitate the future assessment of such trials and will highlight those trials that have been performed suboptimally and whose results may be biased. We have devised a scoring system, based on CONSORT, to assess RCT quality and by reading each original paper in full we have now assessed the quality of trials published from 1966 to 1995. The mean score for trials identified was 7.3 out of a maximum 12 points. No one journal was significantly better than the others. Trials in rhinology are reported better than head and neck oncology trials (mean scores 7.6 and 6.5 respectively). The past 30 years has not seen an improvement in the quality of the trials. The reporting of RCTs in the ENT literature is poor. CONSORT guidelines now exist and trialists are encouraged to adopt them when conducting future clinical trials. PMID- 9747476 TI - Multidirectional observations of the larynx using transurethral rigid endoscopes during direct laryngoscopy. AB - Microscopic direct laryngoscopy (microlaryngoscopy) under general anaesthesia is the optimal method of observing the larynx. However, as microlaryngoscopy does not allow precise observations of the ventricle, inferior surface of the vocal fold and subglottis, multidirectional observations of the larynx using transurethral rigid endoscopes were performed during direct laryngoscopy. This endoscopic technique has been shown to be clinically useful in the diagnosis and treatment of laryngeal lesions. The equipment and methods are introduced herein, and a representative care is presented. PMID- 9747477 TI - Carbon dioxide laser stomaplasty for tracheostomal stenosis. AB - A method of treating tracheostomal stenosis post-laryngectomy is described. The carbon dioxide (CO2) laser is used to fashion and ablate two triangular areas lateral to the stenosed stoma to provide an immediate enlarged stoma for comfortable breathing. This simple procedure is done under local anaesthesia, is almost bloodless, safe and takes just 10 minutes. Over the last five years eight patients underwent this procedure and seven had a satisfactory stoma without the need to use a tracheostomy tube. PMID- 9747478 TI - Multiple myeloma presenting with external ear canal mass. AB - The manifestations of multiple myeloma are protean and related to bony osteolytic lesions, and to medullar and renal insufficiency. We report a patient who presented with otalgia as the inaugural symptom of multiple myeloma. Local irradiation combined with systemic chemotherapy led to the disappearance of the temporal bone mass and the accompanying symptoms. To date, 24 months after the diagnosis, the patient is still in remission. The literature on otological involvement in multiple myeloma is reviewed. Symptoms are non-specific and include hearing loss, tinnitus, dizziness, facial paralysis, and otalgia. The diagnosis of multiple myeloma should be considered in the presence of a temporal bone mass. PMID- 9747479 TI - Hearing preservation despite labyrinthectomy for resection of giant cholesteatoma with middle fossa extension. AB - A case is described of an extensive acquired cholesteatoma of the middle ear cleft which had invaded the middle cranial fossa and produced a mass effect on the temporal lobe. It had also extended into the labyrinth without causing elevation in the bone conduction threshold. Furthermore, even after total bony labyrinthectomy, there was very little elevation in these thresholds. The literature relating to hearing preservation after labyrinthectomy is reviewed. PMID- 9747480 TI - Primary malignant lymphoma in the cerebellopontine angle. AB - Primary malignant lymphoma is an uncommon disease of the central nervous system (CNS). Immuno-compromised patients are at high risk of development of malignant lymphoma. We describe a case of primary CNS lymphoma presenting as a solitary cerebellopontine angle lesion. The patient had undergone extirpation of rectal cancer four years previously. Malignant lymphoma presenting as a cerebellopontine angle mass is extremely rare, with only 10 such cases (seven were primary, and the other secondary) previously reported. PMID- 9747481 TI - Sudden onset aphonia caused by a Japanese-style bath. AB - An 86-year-old man was referred by his family physician to our clinic because of sudden onset aphonia immediately after a Japanese-style bath. On examination, the only abnormality was aphonia, with an otherwise normal physical examination. However, we found vocal fold oedema on laryngeal fibrescopy. For treatment, he was immediately given 30 mg prednisolone intravenously. Twelve hours after injection the oedema had completely disappeared and his aphonia had resolved. The patient was healthy for three months after returning home. However, at the beginning of a cold winter night he again complained of sudden onset hoarseness, after taking a Japanese-style bath. Japanese-style baths are completely different from Western-style baths. There is a temperature difference of almost 30 degrees C between the inside and outside of a bath; the transition may represent a type of physical exercise in elderly and exhausted individuals. This difference could cause a cold or heat-induced allergic reaction. We strongly recommend a laryngeal study in case of sudden onset aphonia. PMID- 9747482 TI - Traumatic laryngocoele. AB - This is the first reported case of a laryngocoele developing after laryngeal trauma. A 26-year-old man sustained a shotgun injury to his larynx. A large number of shotgun pellets was removed from his left vestibular fold. He subsequently developed a left-sided laryngocoele, probably due to fibrosis around the neck of the saccule. The laryngocoele was removed by an external approach. PMID- 9747483 TI - Vocal fold palsy following vinca alkaloid treatment. AB - We report two patients with Hodgkin's disease on chemotherapy, one with vincristine, the other with vinblastine, who were referred to the ENT department with a vocal fold palsy. These drugs are commonly used in the treatment of haematological malignancies, and head and neck sarcomas. Examination showed a unilateral immobile fold. No causative local pathology was identified, and they were otherwise thought to be achieving remission, but the development of a fold palsy suggested a poor response to treatment. One patient's palsy resolved following completion of chemotherapy, but the other patient's persists, despite an otherwise good response to the treatment. The vinca alkaloids are neurotoxic, usually causing a peripheral neuropathy, but cranial mononeuropathies are a rare side-effect. A total of 27 previous cases of vocal fold palsy have been reported with the use of the vinca alkaloids, and such a lesion should not be assumed to be due to advancing lymphoma. PMID- 9747484 TI - Laryngeal aspergillosis in an acquired immunodeficiency syndrome patient. AB - Aspergillosis is a rare infection. It varies considerably in its virulence depending on whether or not the patient affected is immunodepressed. The serious form is characterized by vascular invasion with haematogenic dissemination, tissue infarction and necrosis. The most often affected site is the lung. A laryngeal localization is exceptional and usually results from dissemination from a lower airways portal of entry. Diagnosis is difficult: cultures are only positive in 50 per cent of cases, while in 60 per cent of cases the characteristic histological features reminiscent of sprigs of mistletoe are only found on postmortem examination. In view of the high morbidity and mortality in acquired immunodeficiency syndrome (AIDS) sufferers, this condition must be treated early and aggressively. Treatment is based essentially on amphotericin B and itraconazole. PMID- 9747485 TI - Excision and low-dose radiotherapy for refractory laryngeal granuloma. AB - Laryngeal granulomas are uncommon lesions of an inflammatory origin. They are conventionally managed by simple excision with the occasional use of adjuvant treatment depending on the aetiological factors. Unfortunately, recurrences can occur, requiring repeated excision. Some lesions are refractory to this approach and alternative management approaches include excision and immediate adjuvant radiotherapy. The use of radiotherapy in the management of benign disease can be limited by the risk of induction of late malignancy and informed consent of a patient must include an assessment of this risk. We describe a case of refractory laryngeal granuloma successfully treated by excision and immediate radiotherapy in a patient occupationally dependent upon his voice. PMID- 9747486 TI - Polymerase chain reaction in the diagnosis of parotid gland tuberculosis. AB - A parotid gland mass with presenting features of malignancy is a diagnostic and therapeutic challenge. The histological nature of the lesion must be clearly determined before proceeding with facial nerve sacrificing surgery. Although rare, tuberculosis of the parotid gland must be included in the differential diagnosis of a parotid gland mass especially when the social characteristics of the patient suggests a mycobacterial infection. Primary tuberculosis of the parotid gland is generally encountered among populations with a high incidence of pulmonary disease. The difficulty in the differential diagnosis of a parotid gland malignancy may be helped by a high degree of clinical suspicion, since laboratory tests generally do not identify the specific causative organism. This article reports the first case of parotid gland tuberculosis with clinical and radiodiagnostical features simulating malignancy in which the diagnosis was confirmed by the polymerase chain reaction (PCR). PMID- 9747487 TI - Sudden parotid swelling due to spontaneous haemorrhage. AB - This paper presents an interesting case of a severe spontaneous haemorrhage within the parotid gland in an adult. A rapidly enlarging parotid mass with absence of causative trauma, inflammation or vascular abnormality, raised the suspicion of a neoplasm. Ultrasound, computed tomography (CT) and tissue biopsy, however, have shown only changes suggestive of previous haemorrhage and no evidence of malignancy. A 30-year review of the literature revealed no similar case reported previously. PMID- 9747488 TI - Papillary endothelial hyperplasia (Masson's tumour) of the maxillary sinus. AB - In 1923, Masson described a neoplastic process consisting of papillary hyperplasia of the endothelial cells, with a consequent obliteration of the vascular lumen, followed later by degenerative changes. Masson coined the term vegetant intravascular haemangioendothelioma, however, these days it is more commonly known as papillary endothelial hyperplasia (PEH), or by the pseudonym, Masson's tumour. Although relatively rare, there are numerous accounts of PEH in the literature, describing its predilection for the head and neck region. Our case report describes the finding of a PEH within the paranasal sinuses, a site not previously mentioned even in the largest of series found on literature search. We will then discuss the relevant histological features of the lesion, and its natural history. PMID- 9747489 TI - Solitary fibrous tumour of the deep soft tissues of the neck. AB - A case of a solitary fibrous tumour arising in the deep soft tissues of the neck is reported. This rare tumour has not previously been described in this site. We discuss the clinical presentation and pathological features. PMID- 9747490 TI - A caution in the use of laryngeal sprays. 1898. PMID- 9747491 TI - Psychophysical evidence for a magnocellular pathway deficit in dyslexia. AB - The relationship between reading ability and psychophysical performance was examined to test the hypothesis that dyslexia is associated with a deficit in the magnocellular (M) pathway. Speed discrimination thresholds and contrast detection thresholds were measured under conditions (low mean luminance, low spatial frequency, high temporal frequency) for which psychophysical performance presumably depends on M pathway integrity. Dyslexic subjects had higher psychophysical thresholds than controls in both the speed discrimination and contrast detection tasks, but only the differences in speed thresholds were statistically significant. In addition, there was a strong correlation between individual differences in speed thresholds and reading rates. These results support the hypothesis for an M pathway abnormality in dyslexia, and suggest that motion discrimination may be a more sensitive psychophysical predictor of dyslexia than contrast sensitivity. PMID- 9747493 TI - Multifocal ERG reveals long distance effects of a local bleach in the retina. AB - To examine the distribution of ERG-activity in the central visual field after local bleaching of the fovea, multifocal electroretinograms were recorded in eight normal volunteers before, during and after recurrent light exposure. During bleaching (90% bleached pigment), the response density (scalar product) of the foveal area (0-2 degrees eccentricity) decreased from 10.7 +/- 3.5 to 4.1 +/- 1.9 nV/degree2 (P < 0.001). The average activity in the extrafoveal macular area was unchanged, while the amplitudes were frequently (in 53 of 54 areas) enhanced at 5 30.5 degrees eccentricity. Here the average response density changed from 3.1 +/- 0.9 to 3.5 +/- 1.0 nV/degree2 (P < 0.001). A fast recovery of foveal responses after cessation of bleaching occurred. Besides a strong decrease of response in the directly bleached area, local bleaching led to enhanced activity mainly 3-27 degrees distant from the bleached area. PMID- 9747492 TI - Chromatic induction: border contrast or adaptation to surrounding light? AB - Chromatic induction from a surrounding light is measured with an additional remote field outside the surround. Chromatic induction from the surround into a central test field is found to be attenuated by a remote inhomogeneous 'checkerboard', composed of squares at two different chromaticities. A uniform remote field, on the other hand, either at the average or at the most extreme chromaticity of the 'checkerboard', has a weaker effect on chromatic induction than the inhomogeneous field, implying that chromatic contrast within the remote region is a critical factor. The complete set of experiments is accounted for by chromatic contrast gain control: chromatic induction, mediated by a neural signal for contrast at the edge of the test, is attenuated by contrast within the remote region. A contrast gain control set by variation in chromaticity over a broad area can contribute to the stable color appearance of surfaces embedded within complex scenes by minimizing chromatic induction from locally adjacent regions. PMID- 9747494 TI - Independent speed-tuned global-motion systems. AB - Several experiments were conducted to investigate the role of speed in global motion processing; the extraction of the direction of motion of a small subset of coherently-moving (signal) dots in a stimulus in which the other (noise) dots move in random directions. The specific aim of the experiments was to determine whether multiple speed-tuned global-motion systems exist. The results of these experiments are: (1) when the signal dots were chosen from a group of dots moving at 1.2 degrees s-1, the speed of additional-noise dots had to be below 4.8 degrees s-1 for them to affect global-motion extraction; (2) the addition of static dots did not impair the extraction of a global-motion signal carried by dots moving at 1.2 degrees s-1; (3) noise dots moving at 1.2 degrees s-1 impaired the extraction of a global-motion signal from dots moving at 10.8 degrees s-1, though not to the same extent as dots moving at a higher speed; and (4) these results were dependent upon speed, not spatial-step size or luminance contrast. These results are interpreted as indicating that global-motion extraction occurs within at least two independent speed tuned systems. One of these systems is sensitive to high speeds and the other to low speeds. PMID- 9747495 TI - Local and global factors affecting the coherent motion of gratings presented in multiple apertures. AB - Using stimuli composed of two independent gratings viewed through multiple apertures, we investigate a number of parameters affecting the integration of locally ambiguous motions into globally coherent motion. In four experiments, we varied local factors (grating spatial frequency, speed, contrast, duty cycle, orientation) and global factors (degree of similarity and common fate between the gratings, and symmetry in the configuration of the grating pattern) and examined their effects on global motion coherence. Our results, confirming accounts offered by previous investigators, indicate that local competition between motion signals generated by contours (ambiguous) and their line terminations (unambiguous) is important in determining global motion coherence in multiple aperture stimuli. Our results also indicate that global factors can affect perceived coherence independently of local motion signals, suggesting the involvement of higher-level motion areas and a role for non-motion processes such as those involved in pattern and form perception. Comparing motion coherence with other two-dimensional (2-D) stimuli (plaids) shows that 2-D multiple-aperture stimuli are not analogous and that coherence models derived from plaid stimuli do not account for the data. PMID- 9747497 TI - Veridical perception of global motion from disparate component motions. AB - Although it is in principle possible to determine the direction of motion of an object by combining the motion of its one-dimensional oriented contours (Fennema CL, Thompson WB. Comput. Graph. Image Processing 1979;9:301-315) there is still much debate on whether human observers can do so. The Intersection Of Constraint (IOC) rule proposed by Adelson and Movshon (Adelson EH, Movshon JA. Nature 1982;300:523-525), although compatible with the veridical object's motion, was challenged by recent psychophysical data obtained with type II plaids or lines moving behind apertures: perceived direction of motion is biased toward the vectorial average of the component motions, rather than in the direction predicted by the IOC rule. Since the velocity predicted by the vectorial rule is inconsistent with the physical velocity, its use leads to the puzzling prediction that the perceived position of a moving object becomes inconsistent with its actual position. In the present paper, the perceived path of a figure defined by its one-dimensional contours and moving behind apertures along a circular trajectory is compared with the discrepant predictions of the IOC and of the Vectorial model. The results show that the perceived path is close to veridical with these stimuli, therefore challenging the idea that the visual system uses a vector averaging rule. PMID- 9747496 TI - Motion detection and directional tuning. AB - A random dot pattern was presented which made two jumps in various directions with a variable delay between them. The jumps occurred at the frame transitions of a 3-frame apparent motion sequence. The variation in detectability with the directional difference and the temporal separation of the jumps allows us to make inferences about directional tuning and the temporal response of the motion detection mechanism. The detectability of a pair of jumps was highest when the delay between the jumps was short and the difference in the jump directions was small. In all cases the data were well fitted with a vector version of the speed energy model earlier proposed by Simpson. The model supposes that the two input vectors are temporally filtered, squared and integrated. Using the model, the autocorrelation function of the motion system's temporal impulse response can be found. This function shows the filter to be lowpass. According to the model, the shape of the threshold or d' locus as a function of the difference in the directions of the two jumps does not show the tuning of a motion mechanism. A tuned mechanism will respond well to a jump in its preferred direction, but less well to any other jump. Instead we show that the apparent tuning evident in the threshold and d' loci is due to the way in which the two jump vectors, each fully recovered, are combined in a vector sum. PMID- 9747498 TI - The optimal motion stimulus: comments on Watson and Turano (1995) AB - Watson and Turano (Vision Research 1995;35:325-336) described experimental research aimed at determining the motion stimulus that the visual system detects best. They reported conflicting results in the determination of the optimal spatial size and they interpreted them as an effect of probability summation. They also reported disagreement with earlier results of Watson et al. (Nature 1983;302:419-422). This study shows (i) that probability summation is not responsible for those results and (ii) that they can be explained as a consequence of the method that was used to search for the optimal stimulus. PMID- 9747499 TI - On the time course of exogenous cueing effects: a commentary on Tassinari et al. (1994) PMID- 9747500 TI - The congenital and see-saw nystagmus in the prototypical achiasma of canines: comparison to the human achiasmatic prototype. AB - We applied new methods for canine eye-movement recording to the study of achiasmatic mutant Belgian Sheepdogs, documenting their nystagmus waveforms and comparing them to humans with either congenital nystagmus (CN) alone or in conjunction with achiasma. A sling apparatus with head restraints and infrared reflection with either earth- or head-mounted sensors were used. Data were digitized for later evaluation. The horizontal nystagmus (1-6 Hz) was similar to that of human CN. Uniocular and disconjugate nystagmus and saccades were recorded. See-saw nystagmus (SSN), not normally seen with human CN, was present in all mutants (0.5-6 Hz) and in the one human achiasmat studied thus far. This pedigree is an animal model of CN and the SSN caused by achiasma or uniocular decussation. Given the finding of SSN in all mutant dogs and in a human, achiasma may be sufficient for the development of congenital SSN and, in human infants, SSN should alert the clinician to the possibility of either achiasma or uniocular decussation. Finally, the interplay of conjugacy and disconjugacy suggests independent ocular motor control of each eye with variable yoking in the dog. PMID- 9747501 TI - Static aspects of accommodation: age and presbyopia. AB - Although the progressive reduction in accommodative amplitude with increased age is well documented, little is known about several other aspects of static or steady-state accommodation to provide a comprehensive assessment of changes related to age and presbyopia. Static components of accommodation (tonic accommodation, depth-of-focus, slope of the stimulus/response function, and accommodative controller gain) were assessed objectively using an infrared (IR) optometer in 30 human subjects aged 21-50 years; depth-of-focus was also determined psychophysically as was accommodative amplitude. Tonic accommodation and the amplitude of accommodation decreased with increased age, whereas the subjective depth-of-focus increased; the other parameters remained unchanged. The decrease in tonic accommodation and amplitude of accommodation was attributed to biomechanical factors, whereas the increase in subjective depth-of-focus was believed to result from increased tolerance to defocus related to the gradual onset of presbyopia. Constancy of the objective depth-of-focus suggested absence of age effects on the neurologic control of reflex accommodation, whereas the lack of systematic change in slope and controller gain provided support for the Hess-Gullstrand theory of accommodation and presbyopia. PMID- 9747502 TI - Surface orientation from texture: ideal observers, generic observers and the information content of texture cues. AB - Perspective views of textured, planar surfaces provide a number of cues about the orientations of the surfaces. These include the information created by perspective scaling of texture elements (scaling), the information created by perspective foreshortening of texels (foreshortening) and, for textures composed of discrete elements, the information created by the effects of both scaling and foreshortening on the relative positions of texels (position). We drive a general form for ideal observers for each of these cues as they appear in images of spatially extended textures, (e.g. those composed of solid 2-D figures). As an application of the formulation, we derive a set of 'generic' observers which we show perform near optimally for images of a broad range of surface textures, without special prior knowledge about the statistics of the textures. Using simulations of ideal observers, we analyze the informational structure of texture cues, including a quantification of lower bounds on reliability for the three different cues, how cue reliability varies with slant angle and how it varies with field of view. We also quantify how strongly the reliability of the foreshortening cue depends on a prior assumption of isotropy. Finally, we extend the analysis to a naturalistic class of textures, showing that the information content of textures particularly suited to psychophysical investigation can be quantified, at least to a first-order approximation. The results provide an important computational foundation for psychophysical work on perceiving surface orientation from texture. PMID- 9747503 TI - Discrimination of planar surface slant from texture: human and ideal observers compared. AB - In order to quantify the ability of the human visual system to use texture information to perceive planar surface orientation, I measured subjects' ability to discriminate planar surface slant (angle away from the fronto-parallel) for a variety of different types of textures and in a number of different viewing conditions. I measured the subjects' discrimination performance as a function of surface slant, field of view size and surface texture structure. I compared the subjects' performance with that of ideal observers derived for each of the available texture cues--texel position, scaling and foreshortening. The results can be summarized by four points: (i) subjects' discrimination performance improves dramatically with increasing surface slant, tracking the performance of the ideal observers; (ii) subjects can integrate texture information over a large range of visual angles; (iii) comparisons between human subjects and ideal observers show that the human observers rely to some degree on foreshortening information; and (iv) similar comparisons show that in using foreshortening information, subjects rely to some extent on a prior assumption of isotropy. PMID- 9747504 TI - The role of neural and optical factors in limiting visual resolution in myopia. AB - The myopic growth process has the potential to modify both the optical and neural performance of the eye. We provide three simple models, based on different types of retinal stretching, to predict changes in neural resolution resulting from axial length increases in myopia. These predictions are compared to visual acuity (VA) measures in 34 subjects with refractive errors ranging from plano to -14 D. Our results show a reduction in VA with increasing myopia but not in a manner predicted by our models. We discuss the relative contribution of optical and neural factors to the reduction in visual resolution in myopia. PMID- 9747505 TI - Does dark adaptation exacerbate diabetic retinopathy? Evidence and a linking hypothesis. AB - The paper reviews evidence that before any change in diabetics' fundi, changes occur to blood flow, ERG and visual functions. In the case of colour vision and contrast sensitivity, the changes are partially reversed by breathing oxygen, and therefore are the result of retinal hypoxia. There are also other evidences that hypoxia is a major factor in the development of diabetic retinopathy (DR). Therefore in diabetics with early retinopathy, but normal photopic vision, functional disturbance might appear in dark adaptation, since in such circumstances, (as shown by Linsenmeier and his colleagues) the already low retinal PO2 markedly decreases. This hypothesis has been tested and results consistent with the hypothesis (and with a number of older reports) have been obtained. The significance of this finding to early DR is discussed, and a mechanism suggested whereby prolonged periods of hypoxia during dark adaptation could generate changes in retinal capillaries. Such periods occur each night, and their elimination in diabetics could be therapeutic. PMID- 9747507 TI - Inhibition of bFGF activity by complement C1s: covalent binding of C1s with bFGF. AB - The first complement component C1s formed large aggregates with bFGF when bFGF and C1s were incubated at 37 degrees C overnight. Under non-reducing conditions, a part of the aggregates did not penetrate into 5% polyacrylamide gel in the presence of SDS, and the rest penetrated into 5% gel but not into 12% gel. The aggregates were dissociated into monomers by reducing with 2-mercaptoethanol. Both active and inactive C1s formed aggregates with bFGF. In addition, a portion of bFGF was degraded by active C1s but not by inactive C1s. Aggregates were not formed when 2-mercaptoethanol (2 mM) was added to the incubation mixture. After the incubation with C1s the growth-stimulating activity of bFGF was measured by using human umbilical vein endothelial cells (HUVEC) as indicator cells. The aggregate formation between C1s and bFGF significantly reduced the activity of bFGF. PMID- 9747506 TI - Oxidized low-density lipoprotein decreases the induced nitric oxide synthesis in rat mesangial cells. AB - Recently, the close relation between oxidized low density lipoprotein (Ox-LDL) and the progression of glomerular injury has been demonstrated. The nitric oxide (NO) pathway in glomerular mesangial cells may be a potential target for the adverse effects of Ox-LDL in the development of glomerular injury. In this study, we treated cultured rat mesangial cells (RMC) with Fe(2+)-oxidized LDL and then stimulated the cells with lipopolysacharride (LPS, 10 micrograms ml-1). The LPS induced NO production, assessed by NO2-concentrations in cultured supernatants, decreased from 7.83 nmol per 10(6) cells in control to 4.00 nmol per 10(6) cells and 1.67 nmol per 10(6) cells in RMC preincubated with Ox-LDL at 20 micrograms ml 1 and 40 micrograms ml-1, respectively (P < 0.01). Native LDL had no significant effects on LPS-induced NO production. Using the reverse transcription-polymerase chain reaction (RT-PCR) technique, we could not detect significant alteration of inducible NO synthase (iNOS) mRNA levels in RMC preincubated with Ox-LDL. Our results suggest that Ox-LDL decreases induced NO production in RMC, which may contribute to the adverse effects of Ox-LDL in progressive glomerular injury. The mechanisms of this decrease may not involve changes of iNOS genic transcription. PMID- 9747508 TI - Investigation of the effect of theophylline administration on total, free, short chain acyl and long-chain acyl carnitine distributions in rat renal tissues. AB - This study is conducted to investigate the effect of oral theophylline administration on total (TC), free (FC), short-(SC), long-chain acyl (LC), acyl (AC) carnitine distributions as well as the ratio of acyl to free carnitine (AC/FC) in rat renal tissues. Theophylline was administrated at 100 mg kg-1 body weight day-1, and effects were monitored after a treatment period that lasted between 1 week and 5 weeks. The results indicated that theophylline administration leads to significantly higher concentrations of TC, FC, SC, L and AC in renal tissues as compared to those of control and placebo groups (P < 0.001). Moreover, the ratio of AC/FC was significantly increased (P < 0.001) as compared to either control or placebo groups. These changes may result from theophylline-enhanced mobilization of lipids from adipose tissues, which consequently stimulates an increased carnitine transport into the renal tissues to form acylcarnitines for subsequent beta-oxidation inside the renal mitochondria. PMID- 9747509 TI - Mitochondrial sensitivity to AZT. AB - The possibility of tissue-specific effects regarding mitochondrial sensitivity to AZT was evaluated in this study. When mitochondria isolated from liver, kidney, skeletal and cardiac muscle were oxidizing glutamate, a dose-dependent inhibition by AZT of state 3 respiration was observed; using succinate as substrate the inhibition occurred only in skeletal and cardiac muscle mitochondria. The same results were obtained with FCCP-uncoupled mitochondria. NADH oxidase of intact and disrupted mitochondria, isolated from all four tissues was strongly inhibited. Succinate oxidase activity was inhibited by AZT only in intact mitochondria from skeletal and cardiac muscles, suggesting the involvement of succinate transport systems. Similarly, inhibition by the drug of the hydrolytic activity of H(+)-ATPase was observed only in mitochondria of these tissues. These effects taken together, indicate a tissue/carrier-specific inhibition in vitro, although its precise mechanism requires further research. PMID- 9747510 TI - Oxygen tension regulates heme oxygenase-1 gene expression in mammalian cell lines. AB - The gene expression of heme oxygenase-1 (HO-1) was studied in mammalian cell lines exposed to hyperoxia. Northern blot analysis demonstrated that hyperoxic exposure increased the HO-1 mRNA levels in various types of cells, including human hepatoma (HepG2) cells. This increase was time- and dose-dependent, and reversible. The HO-1 mRNA levels in HepG2 cells were increased to 2.3- and 4.2 fold of the control by hyperoxic exposure of 6 and 23 h, respectively. Cycloheximide and actinomycin D inhibited the increases in the HO-1 mRNA level produced by hyperoxia, indicating that response to hyperoxia is dependent on de novo protein synthesis and mRNA transcription. Antioxidants, desferrioxamine (DES) and o-phenanthroline (OP) partially inhibited the HO-1 mRNA elevation by hyperoxia. In addition to hyperoxia, sodium arsenite (NaAsO2), cadmium chloride (CdCl(2)) and hydrogen peroxide (H2O2), which are reactive oxygen intermediates (ROI) generators, increased the HO-1 mRNA level by 11-, 22- and 2.5-fold, respectively. OP, an antioxidant and a bivalent metal chelator, blocked the HO-1 mRNA elevation induced either by hyperoxia or by the three ROI generators. In contrast to OP, N-acetylcysteine (NAC), an antioxidant and membrane-permeable reducing reagent, enhanced the HO-1 mRNA elevation induced by hyperoxia, although NAC inhibited the mRNA elevation induced by NaAsO2, CdCl2 and H2O2. These results indicate that oxygen tension regulates HO-1 gene expression and suggest that hyperoxia-specific and redox-sensitive regulators may be involved in hyperoxia mediated HO-1 gene expression. PMID- 9747511 TI - The effect of Walker-256 tumour development upon Kupffer cell metabolism. AB - The liver plays a central role in the establishment and maintenance of the cachectic state in rats bearing extra-hepatic tumours. Kupffer cells, which as macrophages, show a strong relationship between metabolism and function could be involved in the alterations observed in the disruption of many functions of the organ as a whole. To assess whether the metabolic/functional pattern of Kupffer cells was altered by cachexia we have investigated the utilization of glucose, glutamine and palmitate by the cells from tumour-bearing and control rats. We have found an enhanced utilization of the three substrates by the cells from tumour-bearing rats as compared with controls, which was related to greater energy production through the Krebs cycle and enhanced production of precursors for the synthesis of the many substances the cells secrete when activated. The use of palmitate as substrate was also augmented in these cells, in the opposition to the observation in stimulated peritoneal macrophages. The availability of palmitate however, was not associated with a reduction of glucose or glutamine consumption. The cycle of interconversion, free fatty acids/triacyglycerol in Kupffer cells from tumour-bearing rats was also found to be increased, as was hydrogen peroxide production. Taken together the results suggest an increased utilization of substrates for both energy production and for synthetic processes (e.g. NADPH for hydrogen peroxide production). PMID- 9747512 TI - Content of dolichol and retinol in isolated rat non-parenchymal liver cells. AB - The liver sinusoids, that are considered as a functional unit, harbour four types of sinusoidal cells (Ito, Kupffer, endothelial and pit cells). Dolichol content has been determined in many tissues and subcellular compartments, alteration has been reported in many types of liver injury, but until now no data are available on its content in every type of sinusoidal non-parenchymal liver cells. Dolichol and retinol metabolism might intersect in their traffic in biological membranes. Intercellular as well as intracellular exchange of retinoids is an essential element of important processes occurring in liver cells. It has been suggested that the role of dolichol, besides being a carrier of oligosaccharides in the biosynthesis of N-linked glycoproteins, may be to modify membrane fluidity and permeability, and facilitate fusion of membranes. Dolichol in the membrane is intercalated between the two-halves of the phospholipid bilayer, but its exact disposition is not known and the movement and distribution of retinoid in membranes may vary with the geometry of the membranes. Therefore the aim of this study is to obtain a global understanding of the sinusoidal system regarding dolichol and retinol content in each type of isolated rat liver sinusoidal cell, in normal conditions and after vitamin A administration. The information that can be drawn from the present results is that with normal vitamin A status of the animal, the dolichol content is almost uniform in all liver cells. After vitamin A supplementation, a great increase of dolichol, together with the known increase of retinol, can be measured only in a subpopulation of the Ito cells, the Ito-1 subfraction. Therefore in the cells that are present in the hepatic sinusoid, different pools of dolichol may have separate functions. Because retinol traffic among cells, membranes and plasma still remains to be fully understood, roles of dolichol in the exchange of vitamin A among sinusoidal liver cells are discussed. PMID- 9747513 TI - CD36 mediates binding of soluble thrombospondin-1 but not cell adhesion and haptotaxis on immobilized thrombospondin-1. AB - In this study, we examined the binding of soluble TSP1 (and ox-LDL) to CD36 transfected cells and the mechanisms by which immobilized TSP1 mediated attachment and haptotaxis (cell migration towards a substratum-bound ligand) of these transfected cells. CD36 cDNA transfection of NIH 3T3 cells clearly induced a dramatic increase in binding of both soluble [125I]-TSP1 and [125I]-ox-LDL to the surface of CD36-transfected cells, indicating that there was a gain of function with CD36 transfection in NIH 3T3 cells. Despite this gain of function, mock- and CD36-transfected NIH 3T3 cells attached and migrated to a similar extent on immobilized TSP1. An anti-TSP1 oligoclonal antibody inhibited CD36 transfected cell attachment to TSP1 while function blocking anti-CD36 antibodies, alone or in combination with heparin, did not. A series of fusion proteins encompassing cell-recognition domains of TSP1 was then used to delineate mechanisms by which NIH 3T3 cells adhere to TSP1. Although CD36 binds soluble TSP1 through a CSVTCG sequence located within type 1 repeats, 18,19CD36 transfected NIH 3T3 cells did not attach to immobilized type 1 repeats while they did adhere to the N-terminal, type 3 repeats (in an RGD-dependent manner) and the C-terminal domain of TSP1. Conversely, Bowes melanoma cells attached to type 1 repeats and the N- and C-terminal domains of TSP1. However, CD36cDNA transfection of Bowes cells did not increase cell attachment to type 1 repeats compared to that observed with mock-transfected Bowes cells. Moreover, a function blocking anti-CSVTCG peptide antibody did not inhibit the attachment of mock- and CD36 transfected Bowes cells to type 1 repeats. It is suggested that CD36/TSP1 interaction does not occur upon cell-matrix adhesion and haptotaxis because TSP1 undergoes conformational changes that do not allow the exposure of the CD36 binding site. PMID- 9747514 TI - Aerobic microbial degradation of aromatic sulfur-containing compounds and effect of chemical structures. AB - Batch data of aerobic microbial degradation rate constants Kb of phenylthio, phenylsulfinyl and phenylsulfonyl acetates have been determined, and the qualitative relationships between their Kb and chemical structures were analyzed. The phenylthio acetates were most subject to microbial transformation, followed by the phenylsulfonyl acetates. The compounds with a methoxy group were easier degraded than those with a isopropoxy group. The nitro-group and chloro-group on benzene were shown to lower the biodegradability, while the nitro-group at the para-position had stronger side effect on degradation than at the ortho-position. PMID- 9747515 TI - A qualitative field method for monitoring pesticides in the edge-of-field runoff. AB - A field method is described, which allows the qualitative estimation of pesticide contamination in the edge-of-field runoff. The method employs cheap and easy-to use runoff sampling bottles, which were installed in an agricultural stream catchment over a period of three growing seasons. During this time 18 runoff events were detected, in nine of which insecticide contamination was measured (maximum concentrations: lindane 0.7 microgram l-1 and 12.7 micrograms kg-1, parathion 20 micrograms l-1 and 728 micrograms kg-1, fenvalerate 18.4 micrograms l-1 and 924 micrograms kg-1). These insecticides were detected mainly as particle bound chemicals. On about 80% of the occasions the presence or absence of runoff measured in the field was in agreement with a simulation of runoff presence or absence using the runoff model KINEROS. PMID- 9747516 TI - Characterization of three major toxaphene congeners in arctic ringed seal by electron ionization and electron capture negative ion mass spectrometry. AB - Three environmentally significant chlorinated bornane (CHB) congeners were extracted from Arviat ringed seal blubber and identified by using gas chromatography/mass spectrometry (HRGC/HRECNIMS (CH4), low resolution EIMS, and linked field scanning). They are referred to as TS2 (Parlar#39, B8-531) [2-exo,3 endo,5-exo,6,6,8b,9c,10c (or 10a)-octachlorobornane], TS3 (Parlar#40, B8-1414) [2 endo,3-exo,5-endo,6-exo,8c,9b,10a,10c (or 10b)-octachlorobornane] and TS4 (Parlar#42, Toxicant A, B8-806/809) [2-exo,3-endo,6,6,8b,8c,9c,10c (or 10a) octachlorobornane/2-exo,3-endo,6,6,8b,9b,9c,10a (or 10b)-octachlorobornane]. This is the first time Toxicant A, known to be the most toxic CHB congener in technical toxaphene, has been found in any significant concentration in a marine mammal. PMID- 9747517 TI - Toxicological evaluation of constructed wetland habitat sediments utilizing Hyalella azteca 10-day sediment toxicity test and bacterial bioluminescence. AB - A toxicological evaluation was conducted on wetland habitats created as a result of run-off from agricultural areas. These temporary wetlands were created by using drop pipes as a means of reducing erosional cutting in agricultural fields. Toxicity bioassays utilizing bacterial bioluminescence and Hyalella azteca were used to assess sediment pore water and whole sediment, respectively. Inhibition of bacterial bioluminescence was initially used to determine relative toxicities of pore water from ten wetland sites. Constructed wetland sites were compared to the University of Mississippi Biological Field Station, a relatively pristine reference site. The H. azteca ten day sediment toxicity test was utilized to assess sediment from four selected sites using survival and growth as toxicological endpoints. Results from the toxicological evaluation, along with extensive ecological evaluations, were used to assess the best approach for implementation of temporary wetland habitats with existing agricultural practices. PMID- 9747518 TI - The assimilation of contaminants from suspended sediment and algae by the zebra mussel, Dreissena polymorpha. AB - Since their invasion into the Great Lakes, zebra mussels, Dreissena polymorpha, have increased the water clarity in Lake St. Clair and Lake Erie due to their extensive particle filtration. Because these particles contain sorbed contaminants, the potential for contaminant accumulation from both suspended sediment and algae were examined. Sediment or algae were dosed with selected radiolabeled polycyclic aromatic hydrocarbon congeners and/or hexachlorobiphenyl (HCBP). Assimilation efficiencies were measured and depended on food quality. Zebra mussels, 17 +/- 2 mm long, assimilated 58.3 +/- 13.5% of the pyrene and 44.7 +/- 5.8% of the benzo(a)pyrene (BaP) from sediment particles with a particle clearance rate of 493-897 ml/g tissue/h. However, assimilation efficiencies were 91.7 +/- 3.7% for pyrene, 91.9 +/- 1.4% for BaP, 96.6 +/- 1.4% for chrysene, and 97.7 +/- 0.5% for HCBP from suspended algae. Algal particle clearance rates for the mussels ranged from 47-143 ml/g tissue/h. Thus, zebra mussels efficiently accumulated non-polar contaminants sorbed to algae, while a smaller fraction of the sediment-associated contaminant was bioavailable. Furthermore, the contaminants sorbed onto suspended sediment particles were quickly removed from the water and deposited as pseudofeces. The pseudofeces production was positively correlated with filtration rate and suspended particle concentrations. PMID- 9747519 TI - Atrazine and metolachlor residues in Brookston CL following conventional and conservation tillage culture. AB - Atrazine and metolachlor are extensively used in Ontario, Canada for control of broadleaf weeds and annual grasses in corn. Conservation tillage may alter the physical and biological environment of soil affecting herbicide dissipation. The rate of dissipation of these two herbicides in soil from conventional, ridge and no-tillage culture was followed. Herbicide dissipation was best described by first order reaction kinetics. Half life, the time for herbicide residues to dissipate to half their initial concentration, was unaffected by tillage. Half life for atrazine and metolachlor was similar and ranged from 31 to 66 d. The rate of dissipation decreased in dry years when soil moisture content was low. In a dry year, herbicide residues during the growing season were significantly greater on ridge tops than in the other tillage treatments. However, after harvest no differences in herbicide residues were detected among tillage treatments. Residues of atrazine (6 to 9% of applied) and metolachlor (4 to 6%) were detected in soil before planting a year after application. De-ethyl atrazine, the primary degradation product of atrazine, increased in concentration during the growing season with the greatest concentrations measured at harvest and in years when atrazine dissipated fastest. De-ethyl atrazine one year after application accounted for about 12% of the remaining triazine residue. These herbicide residues would not be phytotoxic to subsequent crops but are a potential source for leaching to ground and surface waters. PMID- 9747520 TI - Organochlorine pesticides and polychlorinated biphenyl congeners in wild terrestrial mammals and birds from Chubu region, Japan: interspecies comparison of the residue levels and compositions. AB - In order to understand the residue levels of organochlorine compounds (OCs) and their accumulation patterns in wildlife inhabiting Chubu region, Japan, the concentrations of polychlorinated biphenyls (PCBs), hexachlorocyclohexane isomers (HCHs), DDT compounds (DDTs) and hexachlorobenzene (HCB) were measured in 8 species of terrestrial mammals and 10 species of birds. In view of feeding habits, the contamination levels of OCs were found to be higher in omnivorous mammals than in herbivorous ones, and in fish-eating ones and raptores than in omnivorous birds. In fox and dog, PCB-180 (2, 2', 3, 4, 4', 5, 5' heptachlorobiphenyl) was the most dominant PCB congener, while in the other species PCB-153 (2, 2', 4, 4', 5, 5'-hexachlorobiphenyl) was the most persistent. The ratios of lower chlorinated PCB congeners (tri- to tetra-) to total PCBs were larger in fish-eating birds than in the other birds. The results indicate that the compositions of PCB congeners would reflect the differences of feeding habits and xenobiotic metabolizing systems among each species. PMID- 9747521 TI - Responses of the red blood cells from two high-energy-demand teleosts, yellowfin tuna (Thunnus albacares) and skipjack tuna (Katsuwonus pelamis), to catecholamines. AB - In fishes, catecholamines increase red blood cell intracellular pH through stimulation of a sodium/proton (Na+/H+) antiporter. This response can counteract potential reductions in blood O2 carrying capacity (due to Bohr and Root effects) when plasma pH and intracellular pH decrease during hypoxia, hypercapnia, or following exhaustive exercise. Tuna physiology and behavior dictate exceptionally high rates of O2 delivery to the tissues often under adverse conditions, but especially during recovery from exhaustive exercise when plasma pH may be reduced by as much as 0.4 pH units. We hypothesize that blood O2 transport during periods of metabolic acidosis could be especially critical in tunas and the response of rbc to catecholamines elevated to an extreme. We therefore investigated the in vitro response of red blood cells from yellowfin tuna (Thunnus albacares) and skipjack tuna (Katsuwonus pelamis) to catecholamines. Tuna red blood cells had a typical response to catecholamines, indicated by a rapid decrease in plasma pH. Amiloride reduced the response, whereas 4,4'diisothiocyanatostilbene-2,2' disulphonic acid enhanced both the decrease in plasma pH and the increase in intracellular pH. Changes in plasma [Na+], [Cl-], and [K+] were consistent with the hypothesis that tuna red blood cells have a Na+/H+ antiporter similar to that described for other teleost red blood cells. Red blood cells from both tuna species were more responsive to noradrenaline than adrenaline. At identical catecholamine concentrations, the decrease in plasma pH was greater in skipjack tuna blood, the more active of the two tuna species. Based on changes in plasma pH, the response of red blood cells to catecholamines from both tuna species was less than that of rainbow trout (Oncorhynchus mykiss) red blood cells, but greater than that of cod (Gadus morhua) red blood cells. Noradrenaline had no measurable influence on the O2 affinity of skipjack tuna blood and only slightly increased the O2 affinity of yellowfin tuna blood. Our results, therefore, do not support our original hypothesis. The catecholamine response of red blood cells from high-energy-demand teleosts (i.e., tunas) is not enhanced compared to other teleosts. There are data on changes in cardio-respiratory function in tunas caused by acute hypoxia and modest increases in activity, but there are no data on the changes in cardio-respiratory function in tunas accompanying the large increases in metabolic rate seen during recovery from exhaustive exercise. However, we conclude that during those instances where high rates of O2 delivery to the tissues are needed, tunas' ability to increase cardiac output, ventilation volume, blood O2 carrying capacity, and effective respiratory (i.e., gill) surface area are probably more important than are the responses of red blood cells to catecholamines. We also use our data to investigate the extent of the Haldane effect and its relationship to blood O2 and CO2 transport in yellowfin tuna. Yellowfin tuna blood shows a large Haldane effect; intracellular pH increases 0.20 units during oxygenation. The largest change in intracellular pH occurs between 40-100% O2 saturation, indicating that yellowfin tuna, like other teleosts, fully exploit the Haldane effect over the normal physiological range of blood O2 saturation. PMID- 9747523 TI - Energetic cost of hovering flight in a nectar-feeding bat measured with fast response respirometry. AB - Hover-feeding glossophagine bats provide, in addition to the hummingbirds, a second vertebrate model for the analysis of hovering flight based on metabolic measurement and aerodynamic theory. In this study, the power input of hovering Glossophaga soricina bats (11.9 g) was measured by standard respirometry and fast response (< 0.2 s) oxygen analysis. Bats needed 5-7 s after a rest-to-flight transition to return to a respiratory steady state. Therefore, only hovering events preceeded by a 7-s flight interval were evaluated. VO2 during hovering fluctuated with a frequency of 3-5 Hz, which corresponded in frequency to the licking movement of the tongue. During hovering, bats often may have hypoventilated as indicated by reduced VO2 and a respiratory exchange ratio (RER) well below the steady-state value of 1. Steady-state oxygen consumption (and derived power input) during hovering was estimated to be 27 (25-29) ml O2 g-1 h-1 (158 W kg-1 or 1.88 W) in the 11.9-g bats as indicated by three independent findings: (1) VO2 was 26 ml O2 g-1 h-1 after 6.5 s of hovering, (2) the mean RER during single hovering events was at its steady-state level of 1 only at oxygen uptake rates of 25-29 ml g-1 h-1, and (3) when the oxygen potentially released from estimated oxygen stores was added to the measured oxygen uptake, the upper limit for oxygen consumption during hovering was found to be 29 ml O2 g-1 h-1. Hovering power input was about 1.2 times the value of minimum flight power input (Winter and von Helversen 1998) and thus well below the 1.7-2.6 difference in power output postulated by aerodynamic theory (Norberg et al. 1993). Mass specific power input was 40% less than in hummingbirds. Thus, within the possible modes of hovering flight, Glossophaga bats seem to operate at the high-efficiency end of the spectrum. PMID- 9747522 TI - Short-day enhancement of immune function is independent of steroid hormones in deer mice (Peromyscus maniculatus). AB - The effects of photoperiod and steroid hormones on immune function were assessed in male and female deer mice (Peromyscus maniculatus). In experiment 1, male deer mice were castrated, castrated and given testosterone replacement, or sham operated. Half of each experimental group were subsequently housed in either long (LD 16:8) or short days (LD 8:16) for 10 weeks. Short-day deer mice underwent reproductive regression and displayed elevated lymphocyte proliferation in response to the T-cell mitogen concanavalin A, as compared to long-day mice. In experiment 2, female deer mice were ovariectomized, ovariectomized and given estrogen replacement, or sham-operated. Animals from each of these experimental groups were subsequently housed in either LD 16:8 or LD 8:16 for 10 weeks. Short day deer mice underwent reproductive regression and displayed reduced serum estradiol concentrations and elevated lymphocyte proliferation in response to concanavalin A, as compared to long-day mice. Surgical manipulation had no effect on lymphocyte proliferation in either male or female deer mice. Neither photoperiod nor surgical manipulation affected serum corticosterone concentrations. These results confirm that both male and female deer mice housed in short days enhance immune function relative to long-day animals. Additionally, short-day elevation in splenocyte proliferation appears to be independent of the influence of steroid hormones in this species. PMID- 9747524 TI - Effects of divided attention on encoding and retrieval processes in human memory: further support for an asymmetry. AB - Despite a tradition in cognitive psychology that views encoding and retrieval processes in human memory as being similar, F. I. M. Craik, R. Govoni, M. Naveh Benjamin and N. D. Anderson (1996) have recently shown that notable differences exist between the 2 when divided-attention manipulations are used. In this article, the authors further examined this asymmetry by using several manipulations that changed task demands at encoding and retrieval. The authors also used a secondary-task methodology that allowed a microlevel analysis of the secondary-task costs associated with encoding and retrieval. The results illustrated the resiliency of retrieval processes to manipulations involving different task demands. They also indicated different loci of attention demands at encoding and retrieval. The authors contend that whereas encoding processes are controlled, retrieval processes are obligatory but do require attentional resources for their execution. PMID- 9747525 TI - A decrement-to-familiarity interpretation of the revelation effect from forced choice tests of recognition memory. AB - In the revelation effect, the probability of labeling a target or a lure as "old" on item recognition tests increases if just prior to their recognition judgment, participants first identify a disguised version of the test item. The same occurs with interpolated tasks that occur just prior to a recognition judgment if the task shares constituents with the test items. One explanation of this test bias is an increased feeling of familiarity that comes from the identification stage preceding the recognition judgment (e.g., D. C. LeCompte, 1995; C. R. Lou, 1993). This study's finding in 4 experiments that 2-alternative forced-choice recognition either yields no effects of revelation or an "antirevelation" effect, even when both items were studied or nonstudied, is incongruent with this explanation. The authors argue that revelation decrements familiarity, and this results in a more liberal criterion shift. They also argue that their theory is more consistent with previous empirical data. PMID- 9747526 TI - On the recollection of specific- and partial-source information. AB - Memory judgments can be based on information that is more or less specific with respect to the source of an item. The authors introduce a procedure and multinomial model for measuring specific- and partial-source information. In 2 experiments, participants heard words spoken by 4 different voices: 2 male voices and 2 female voices. During the test, participants were required to remember who spoke the test items (e.g., Male 1, Male 2, Female 1, Female 2, or new word). Participants often remembered information about the gender of the source (i.e., partial-source information) when they did not remember information that identified the source itself (i.e., specific-source information). Dividing attention during retrieval impaired participants' memory for specific-source information (i.e., voice information) but did not affect memory for partial source information (i.e., gender information). PMID- 9747527 TI - The integration of object properties. AB - Is a complex object learned as a whole from the onset of learning, or is property recall fragmented and only integrated with repeated experience? The authors studied the recall of 3-property visual objects and found that even when property recall was low, recall coherence was high. Properties not organized as visual objects did not show high recall coherence. In addition, the authors failed to find evidence that spatial location, spatial proximity, spatial distinctiveness, distinctive motion, or perceptual grouping induced by common motion affected the integration of properties. Unitariness, temporal contiguity, and intention, however, were factors that influenced memory coherence. The authors concluded that integration occurs when properties are encoded as the constituents of a cognitive structure. PMID- 9747528 TI - Item repetition in short-term memory: Ranschburg repeated. AB - In serial recall from short-term memory, repeated items are recalled well when close together (repetition facilitation), but not when far apart (repetition inhibition; the Ranschburg effect). These effects were re-examined with a new scoring scheme that addresses the possibility that repetitions are distinct tokens in memory. Repetition facilitation and repetition inhibition proved robust, and were shown to interact with the temporal grouping of items (Experiment 1), which affected the probability of detecting repetition (Experiments 2A and 2B). It is argued that detection of a repetition is necessary for repetition facilitation, attributable to the tagging of immediate repetition, whereas the failure to detect or remember a repetition results in repetition inhibition, attributable to an automatic suppression of previous responses and a bias against guessing repeated items (Experiment 3). The findings are discussed in relation to models of short-term memory and the phenomenon of repetition blindness. PMID- 9747529 TI - Surprise and schema strength. AB - Through 4 experiments, the author investigated the effects of stimuli discrepant with schemas of varying strength on 3 components of surprise: the interruption of ongoing activities (indexed by response time increase), the focusing of attention on the schema-discrepant event (indexed by memory performance), and the feeling of surprise (indexed by self-reports). Response times were consistently found to increase with schema strength. This effect was attributed to the increasing difficulty of schema revision. In contrast, memory for the schema-discrepant event was not affected by schema strength, supporting the hypothesis that schema discrepant stimuli are stored in memory with a distinct tag. Finally, self reports of surprise intensity varied with schema strength only if they were made immediately after the surprising event without any intervening questions, suggesting that self-reports of surprise are highly susceptible to memory distortions. PMID- 9747530 TI - Updating a situation model: a memory-based text processing view. AB - Previous research (J. E. Albrecht & E. J. O'Brien, 1993) demonstrated that readers were aware of an inconsistency between an earlier described characteristic of a protagonist and a subsequent target action carried out by the protagonist. In a series of 5 experiments, a qualification was added to the described characteristic that restricted the conditions under which the characteristic was operative. According to the here-and-now view of mapping, readers should use this qualification to maintain a fully updated model of the protagonist in active memory and should not experience comprehension difficulty when reading the target action. In contrast, according to the memory-based text processing view, the qualification would not be part of the active discourse model. Instead, it would be reactivated when the target action was read. Thus, readers should still experience comprehension difficulty. Results of all 5 experiments were consistent with the memory-based text processing view. PMID- 9747531 TI - Retrieval from temporally organized situation models. AB - Time is an important part of establishing situations in the world. As such, temporal information should be reflected in the organization of information into situation models. This article reports 3 experiments that explore whether people will integrate sets of related facts into situation models in a time-based fashion. People memorized lists of facts and then took a speeded recognition test. A retrieval interference methodology was used to assess whether they had integrated the facts into situation models. The presence of interference indicated a lack of integration. In contrast, a marked reduction or an absence in interference indicated integration. In 2 experiments, time-based integration was observed when common time periods were referred to by either events (e.g., "when the camera flashed") or verb tense (i.e., past, present, and future). A 3rd experiment demonstrated that common time periods alone are not sufficient; the information must be allowed to occur potentially within the same situation. PMID- 9747532 TI - Verbs and nouns are organized and accessed differently in the mental lexicon: evidence from Hebrew. AB - A masked priming paradigm was used to examine the role of the root and verbal pattern morphemes in lexical access within the verbal system of Hebrew. Previous research within the nominal system had showed facilitatory effects from masked primes that shared the same root as the target word, but not when the primes shared the word pattern (R. Frost, K. I. Forster, & A. Deutsch, 1997). In contrast to these findings, facilitatory effects were obtained for both roots and word patterns in the verbal system. In addition, verbal pattern facilitation was obtained even when the primes were pseudoverbs consisting of illegal combinations of roots and verbal patterns. Significant priming was also found when the primes and the targets contained the same root. The results are discussed with reference to the factors that may determine the lexical status of morphological units in lexical organization. A model of morphological processing of Hebrew words is proposed. PMID- 9747533 TI - Confidence-accuracy inversions in scene recognition: a remember-know analysis. AB - S. E. Clark (1997) offered a modified signal-detection explanation of the confidence-accuracy inversions observed in E. Tulving's (1981) experiments. In addition to replicating E. Tulving (1981), we had participants make "remember familiar" judgments. Confidence and accuracy dissociated across subjective reports. Response confidence differed only for judgments based on familiarity, whereas accuracy differed only for "remember" responses. S. E. Clark's model does not predict this, nor can it mimic "remember" performance across all conditions. We propose that although "knowing" can be accommodated within an equal variance signal-detection account, "remembering" is governed by contextual constraints that influence the distinctiveness of information upon which participants rely during reports. The current paradigm is a pictorial analogue to H. L. Roediger and K. B. McDermott's paradigm (1995) in that participants claim to explicitly remember thematically related items that were not actually seen during study. PMID- 9747534 TI - Response bias in visual serial order memory. AB - The recency-to-primacy shift represents a major challenge for all theories that attempt to explain the effects of serial order on memory. At short retention intervals, strong recency and no primacy effects occur, but as the retention interval increases, recency is attenuated and primacy increases. In 2 experiments, 24 participants were presented with sets of 4 unfamiliar faces and were asked to state the serial position of a probe face after 0 or 10 s. The predicted recency-to-primacy shift was obtained with accuracy responses. However, the distribution of responses also showed that there was a change in response bias with retention interval. When this was corrected for, the recency-to-primacy shift was eliminated. Response bias is suggested as the underlying cause of the recency-to-primacy shift in this task. PMID- 9747535 TI - Purification and immunological characterization of a recombinant trimethylflavonol 3'-O-methyltransferase. AB - A flavonol O-methyltransferase cDNA clone (pF3'OMT) from Chrysosplenium americanum was expressed in Escherichia coli Top 10 and the recombinant protein was purified to near homogeneity by affinity chromatography on chelation resin and gel filtration on Superose 12 columns. The purified protein was enzymatically active as a 42 kDa monomer and exhibited strict specificity for position 3' of 3,7,4'-trimethylquercetin. In did not accept the mono- or dimethyl analogs, the parent aglycone quercetin or the phenylpropanoids, caffeic and 5-hydroxyferulic acids as substrates; thus indicating its involvement in the later steps of polymethylated flavonol synthesis in this plant. The K(m) values of the enzyme for 3,7,4'-trimethylquercetin as substrate and S-adenosyl-L-methionine as co substrate were 7.2 and 20 microM, respectively. The enzyme activity was strongly inhibited by both Ni2+ and rho-chloromercuribenzoate and was restored by the addition of EDTA or beta-mercaptoethanol, respectively. Antibodies raised against the F3'OMT recombinant protein recognized a protein band migrating at the expected molecular mass of the enzyme on SDS-polyacrylamide gels of protein extracts prepared from various sources. This implies a high degree of structural similarity among these enzymes that is also corroborated by their hydropathy profiles. PMID- 9747536 TI - Allium chemistry: identification of organosulfur compounds in ramp (Allium tricoccum) homogenates. AB - Supercritical fluid (SF) extracts of homogenized ramp (Allium tricoccum Ait.) were separated and characterized with liquid chromatography coupled with atmospheric pressure chemical ionization mass spectrometric identification. The profiles of SF extracts of aqueous homogenates of ramp bulbs from three different seasons and growing regions revealed that the thiosulfinates were major components. In addition, some of the cepaenes (alpha-sulfinyldisulfides) found in extracts of onion juice, as well as allyl containing cepaenes (2-propenyl l-(2 propenylsulfinyl)propyl disulfide), are present in the ramp extracts. The amount of allicin in ramp bulb homogenates ranged from approximately 10% to 50% of that found in extracts of aqueous garlic homogenates. The greater amount of the methyl 1-propenyl thiosulfinates in the ramp extracts relative to that found in the garlic extracts correlates with the flavor characteristics of ramp bulbs. PMID- 9747537 TI - Jasmonate modulates development- and light-regulated alkaloid biosynthesis in Catharanthus roseus. AB - Methyl jasmonate, a chemical inducer of secondary metabolism, has been shown to promote vindoline biosynthesis in developing seedlings, as a result of induction of tryptophan decarboxylase (TDC) and desacetylvindoline 4-hydroxylase (D4H). The present studies suggest that jasmonate-based induction of TDC and D4H activities involves modulation of transcriptional, post-transcriptional and post translational controls. The effects of jasmonate on both enzymes were transient with maximum TDC activity appearing 12 h earlier than that of D4H. Jasmonate treatment of etiolated seedlings neither enhanced TDC activity nor could it replace the light requirement for D4H induction. Jasmonate, therefore, appears to modulate events which are already triggered by developmental and environmental specific controls. Salicylic acid, another chemical inducer of secondary metabolism, was ineffective in activating either TDC or D4H under the experimental conditions used. PMID- 9747538 TI - Differential regulation and distribution of acridone synthase in Ruta graveolens. AB - Cell suspension cultures of Ruta graveolens L. accumulate polyketide metabolites such as acridone alkaloids and flavonoid pigments. Whereas flavonoid synthesis is induced by light, the production of alkaloids can be enhanced in dark-cultured cells by treatment with fungal elicitors. Acridone synthase (ACS) catalyzes the committed condensing reaction of acridone biosynthesis yielding 1,3-dihydroxy-N methylacridone from N-methylanthraniloyl- and malonyl-CoAs. The reaction proceeds in a manner analogous to that of chalcone synthase (CHS) which catalyzes the first committed step in flavonoid biosynthesis and cDNA and protein sequences of Ruta ACS possess a high degree of sequence homology to heterologous CHSs. ACS transcript abundance and specific activity were monitored in cultured R. graveolens cells irradiated either continuously with white light or treated with fungal elicitor over a period of 24 h and found to increase transiently upon elicitor treatment and to decrease upon light irradiation. Immunodetection with a rabbit polyclonal ACS antiserum revealed that the amounts of ACS polypeptide decreased slightly in light-irradiated cells but increased in elicitor-treated Ruta cells. Fluorescence microscopy and tissue print hybridizations were employed to aid in localizing the sites of storage and biosynthesis of acridone alkaloids in Ruta plants. Yellow fluorescing alkaloids were detected particularly in root tissue adjacent to the rhizodermis, but also in the endodermis and vascular tissue of the hypocotyl. ACS transcript abundance in situ followed the same spatial pattern, indicating that the synthesis of acridones likely proceeds at all sites of deposition rather than exclusively in the root. Expression in planta and the induction response of ACS suggest that the alkaloids serve as phytoanticipins or phytoalexins in the defense of Ruta particularly to soil-borne pathogens or as feeding deterrents. PMID- 9747539 TI - Changes in arginine, PAL activity, and nematode behavior in salinity-stressed citrus. AB - De novo arginine biosynthesis has been described as a response of citrus to a range of stresses. It is often noted that stress in plants enhances susceptibility to herbivory and pathogenic attack. Using a citrus and nematode (Tylenchulus semipenetrans) system, the effects of salinity stress on nematode behavior, amino acids (particularly arginine), and phenylalanine ammonia lyase (PAL) activity was investigated. The hypothesis was tested that under salinity stress, citrus grows more slowly and produces arginine in response to high levels of in vivo ammonia, resulting in lower PAL activity and increased susceptibility to nematode attack. After 30 days of high salinity (0.1 M NaCl), plants exhibited a 38% reduction in growth, 35% reduction in PAL activity, and had 54% higher infection rates. PAL activity was inversely correlated (P < or = 0.05) with salinity level and with increase in arginine concentration. PMID- 9747540 TI - Monoterpene biosynthesis in the liverwort Conocephalum conicum: demonstration of sabinene synthase and bornyl diphosphate synthase. AB - (-)-Sabinene is the major monoterpene produced by a European strain of the liverwort Conocephalum conicum. A cell-free extract from in vitro cultured plants catalysed the cyclization of geranyl diphosphate to sabinene. The responsible monoterpene cyclase was partially purified and characterized as an operationally soluble enzyme of M(r) 65,000, with a pH optimum at 7.5 and a requirement for a divalent metal ion as the only cofactor, with Mg2+ preferred. The general properties of the sabinene synthase from C. conicum resemble those of other monoterpene cyclases from gymnosperms and angiosperms. A North American strain of the liverwort produces (+)-bornyl acetate as the major monoterpene and it was demonstrated that bornane-type monoterpenes are derived from geranyl diphosphate in this liverwort, as in higher plants, by the action of bornyl diphosphate synthase. PMID- 9747541 TI - Antimycobacterial evaluation of germacranolides. AB - The minimum inhibitory concentrations (MIC) against Mycobacterium tuberculosis and M. avium of parthenolide, costunolide, 1 (10)-epoxycostunolide and other germacranolide-type sesquiterpene lactones and derivatives were determined by use of a radiorespirometric bioassay. Structure-activity relationship studies with natural and semisynthetic sesquiterpene lactones suggested that the alpha methylene-gamma-lactone moiety is an essential, but not sufficient, structural requirement for significant in vitro activity against M. tuberculosis and M. avium. PMID- 9747542 TI - Muricoreacin and murihexocin C, mono-tetrahydrofuran acetogenins, from the leaves of Annona muricata. AB - Bioactivity-directed fractionation of the leaves of Annona muricata L. (Annonaceae) resulted in the isolation of two new Annonaceous acetogenins, muricoreacin (1) and murihexocin C (2). Compounds 1 and 2 showed significant cytotoxicities among six human tumor cell lines with selectivities to the prostate adenocarcinoma (PC-3) and pancreatic carcinoma (PACA-2) cell lines. PMID- 9747543 TI - Mental health research in general practice: from head counts to outcomes. PMID- 9747544 TI - Palliative terminal care. PMID- 9747545 TI - Palliative care at home: an audit of cancer deaths in Grampian region. AB - BACKGROUND: Ninety per cent of the last year of life of cancer patients is spent at home. Some studies have suggested that care in this setting is often suboptimal. Information on the standard of palliative care delivered at home by general practitioners (GPs) and their teams is limited, and clarification of the problems faced is needed. AIM: To audit the home-based palliative care of patients dying of cancer. METHOD: Matched postal questionnaires were sent to the GPs and nurses of 1086 successive patients dying of cancer in whatever setting in the Grampian region of Scotland some six weeks after the death to establish the professionals' perception of symptom control, communication problems, use of services, and information given to patients and relatives. RESULTS: Response rates were 88.8% for GPs (964 out of 1086) and 87.1% for nurses (325 out of 375 that were passed on to nurses). Two-thirds of patients received palliative care at home. Pain was poorly controlled in 15.7%, and poor control of other symptoms ranged from 13.8% (nausea and vomiting) to 21% (depression and dyspnoea). Communication difficulties were present in 93.7% of cases, although only 5.2% of these were of a major nature. District nurses were involved in 76.7% of cases and Macmillan nurses in 28.0%. Twenty-six per cent of referrals to district nurses were assessed as being late in the course of the illness. Patients were fully informed about the diagnosis in 66.3% of cases and about the prognosis in 55.4%. General practitioners were more likely to report the presence of communication problems between themselves and the patient (when compared with nurses: 43.9% versus 28.0%), more likely to report that patients were 'not at all informed' about self-help groups (57.5% versus 36.3%), and were less likely to report the involvement of occupational therapists (21.8% versus 39.7%). CONCLUSIONS: Levels of reporting of poor symptom control by professionals was much lower than levels reported by relatives in other studies, but there was no difference between the reporting of GPs and nurses. However, a number of areas were identified where care could be enhanced by improved teamwork and further education and training in symptom control, as well as in communication, use of services, and information provision. PMID- 9747546 TI - Nurse triage for house call requests in a Tyneside general practice: patients' views and effect on doctor workload. AB - BACKGROUND: Demand for consultations in primary care has risen recently, necessitating a change in working practices. As part of this process, the possible contribution of practice nurses in the telephone assessment of home visit requests merits attention. AIMS: To survey the views of our patients encountering our nurse triage system for home visit requests, set up in June 1995, and to plot its effect on the routine visiting workload of our doctors and thus their availability at the surgery. METHOD: The outcome of each request was categorized as: doctor to visit (DV), surgery consultation with doctor (SC), nurse advice given and accepted (NA), or call passed to doctor for advice (DA). Frequency data from September 1995 to December 1996 were recovered. Questionnaires for self-completion were sent to all those requesting a routine weekday house call during two four-week periods in 1995 and 1996. RESULTS: Analysable activity data revealed 1764 house call requests, with 41% DV, 18% SC, 24% NA, and 8% DA. In the first survey, 121 questionnaires were sent out and 84 returned (69% response rate) and, in the second, the corresponding figures were 113, 85, and 75%. About 80% of responders reported that they were satisfied with the help received from the nurse. CONCLUSIONS: Nurse triage of house call requests has led to more efficient care for our patients, as we have increased the availability of surgery consultations by reducing the number of house calls made by our general practitioners. PMID- 9747547 TI - Improving general practitioner clinical records with a quality assurance minimal intervention. AB - BACKGROUND: Although good medical records have been associated with good care, there is considerable room for their improvement in general practice. AIM: To improve the quality of general practice medical records at minimal cost. METHOD: A total of 150 randomly sampled general practitioners (GPs) in suburban Brisbane, Australia, were randomized in a controlled trial to receive or not receive an intervention. The intervention consisted of 6 to 12 one-hour monthly meetings when the pairs of GPs assessed samples of each other's medical records using a 12 item instrument. This was developed previously by a process of consensus of general practice teachers. Mean scores of 10 medical records selected at random from before the intervention started and one year later were compared. RESULTS: After the intervention, the increase in the total score (for which the maximum possible was 18) for the intervention GPs (from a baseline of 11.5 to 12.3) was not significantly greater than for the controls (from 11.4 to 11.7). Legibility and being able to determine the doctor's assessment of the consultation were significantly improved. The post-intervention increase of 1.06 (9.3%) of the total scores of the 47% of intervention GPs who complied with the intervention was significantly greater than that for the controls. CONCLUSION: The quality assurance activity improved some components of the quality of GPs' clinical records. However, the improvement was small, and the search for activities for Australian GPs that demonstrate an improvement in the quality of their practice must continue. PMID- 9747548 TI - Palliative terminal cancer care in community hospitals and a hospice: a comparative study. AB - BACKGROUND: Despite palliative care being an accepted role of community hospitals, there is little quantitative evidence of the type of care provided. AIM: To obtain quantitative data comparing palliative cancer care provided in 12 community hospitals in 10 towns (approximately 350 medical beds) and in a consultant-led purpose-built hospice (12 beds). METHOD: Retrospective medical and nursing case note analysis over one year of cancer deaths in the former Exeter Health District. RESULTS: A total of 171 community hospital and 116 hospice casenotes were analysed. Hospice patients had significantly different reasons for admission compared with community hospital patients (P < 0.001), with pain and symptom control being more frequent and terminal nursing care less frequent reasons for admission to the hospice. Community hospital length of stay was significantly longer than hospice length of stay (P = 0.002; mean community hospital stay 16 days, mean hospice stay eight days). Symptoms on admission differed significantly. Drug prescribing on admission and at death and indications of active treatment of symptoms were broadly similar. Community hospital patients received more investigations than hospice patients, linked to the observation that around one in ten community hospital patients were admitted for investigation and active treatment. Community hospital medical notes were significantly less likely to meet minimum quality standards than were hospice notes (81/171 vs. 18/116; P < 0.001), with major deficiencies in the areas of examination, progress reporting, and absence of confirmation of death. CONCLUSIONS: This study confirms the role of community hospitals in palliative terminal cancer care. Differences in care between community hospitals and a hospice have been demonstrated that may reflect either different admission populations to each setting or differences in the way care was delivered. PMID- 9747549 TI - Dying from cancer in community hospitals or a hospice: closest lay carers' perceptions. AB - BACKGROUND: Despite there being around 400 community hospitals in the United Kingdom, there is little published research on the quality of service provided by these hospitals. AIM: To compare the quality of terminal cancer care in community hospitals with a hospice as assessed by patients' closest lay carer (relative or friend). METHOD: Structured interview (or questionnaire based on the interview proforma) with closest lay carers of all patients dying over one year in 12 community hospitals in east Devon and a purpose-built hospice in the city of Exeter. RESULTS: A total of 292 cases (176 in community hospitals and 116 in a hospice) were identified, resulting in 238 carers being eligible for interview or questionnaire survey. Overall, 106 successful interviews and 55 questionnaires were completed, giving a response rate of 67.6%. Carers gave a near unanimous vote of excellence for the total care given by the hospice, while around 40% of carers of patients in community hospitals considered that improvements were possible. Community hospitals attracted more negative comments than hospices, with criticism being directed at problems of communication, lack of nursing staff, and lack of support in bereavement. Carers of hospice patients were significantly more likely to be present at the time of death than those of community hospital patients [45/70 (64%) vs. 31/89 (35%); chi 2 = 13.6, P < 0.001], an observation possibly because nursing staff in community hospitals are less experienced at dealing with terminally ill patients and such hospitals have fewer adequate facilities. CONCLUSIONS: Lay carers indicated great satisfaction with care given in the hospice and less satisfaction with care given in the community hospitals. However, the community hospitals are non-specialist units with far lower levels of trained staff. Improvements in terms of the communication skills of doctors and nurses, specific training for nurses in palliative care, and structured bereavement care could be made without necessarily increasing staffing numbers. PMID- 9747550 TI - Survey of research activity, training needs, departmental support, and career intentions of junior academic general practitioners. AB - BACKGROUND: Recent changes in the organization of the National Health Service have created new roles and responsibilities for academic general practice. Previous work on the constraints and opportunities of a career in academic general practice is largely anecdotal and is often based on the views of more senior members of the profession. AIM: To survey the research activity, perceived level of training, support needs, and career intentions of junior academic general practitioners (GPs). METHOD: A postal, validated, semistructured questionnaire was sent to the 121 eligible junior academic GPs in the academic departments of general practice in the United Kingdom and Dublin. Main outcome measures were 'research activity score', as measured by publications in peer reviewed journals and involvement in research projects, 'training score' devised from 13 skills required for both research and teaching, and perceived level of departmental support assessed by six different support mechanisms. RESULTS: Response rate was 89% (n = 108). Forty-six responders (43%) had no publications. Twenty-five responders (23%) had no principal project. Thirty-nine responders (37%) had a mentor. Research activity appeared to be dependent on sex, having a predominantly research role rather than a full-time teaching role, and a positive perception of academic training (P < 0.05). Increasing departmental 'support scores' and length of time in the department were both significantly associated with more positive perceptions of academic training (P < 0.05). Only 29 (27%) responders wanted to progress to senior positions within academic general practice. CONCLUSION: Training and departmental support and guidance available to junior academics in primary care are perceived as variable and often inadequate. If academic general practice is to thrive, improved academic training is required, such as taught Master's degrees, supervised personal projects or 'apprenticeship' as a co-investigator, and improved methods of departmental support. PMID- 9747551 TI - General practitioners' perceptions of private health screening: too much paper, anxiety, and reassurance. AB - There is no evidence to support the practice of screening consultations that include general physical examinations and batteries of tests; however, many patients may choose, or be sent by their employers, to have private full health screening (FHS). General practitioners (GPs) are routinely sent the results of these screening examinations and are expected to deal with any subsequent care required. GPs recognize some positive aspects of FHS, but in our survey there was a groundswell of dislike for these examinations because of uncertainty about patient benefit (raised anxiety or false assurance) and a potential to irritate the GP. The implications for workload were minimal but resented. GPs would welcome a precise summary of significant findings and for the screening doctor to take greater responsibility for follow-up. PMID- 9747552 TI - Do women with HIV infection consult with their GPs? AB - In a cohort of 106 HIV-positive women, 86 (81%) were registered with a general practitioner (GP) and 71 (83%) had a GP who was aware of their HIV status. GPs were primarily consulted for perceived non-HIV-related problems and prescriptions. This is encouraging. However, primary and secondary care services should aim to increase the proportions of HIV-positive individuals with access to primary care. PMID- 9747553 TI - A review of tonsillectomy for recurrent throat infection. AB - Tonsillectomy is most frequently carried out for recurrent throat infection, but there is uncertainty about its effectiveness. This paper reviews the evidence of its effectiveness obtained from a search of the Cochrane database and MEDLINE for randomized controlled trials comparing tonsillectomy with non-surgical management of recurrent throat infection. The results show that the effectiveness of a procedure such as tonsillectomy, needs to be considered in the light of its adverse effects. Attempts should be made to inform patients about the uncertainty surrounding the procedure. PMID- 9747554 TI - Literature in our medical schools. AB - Despite many relevant benefits, the study of literature has been rejected by medical schools this century. However, the role of literature and the arts is coming to the fore again in many branches of medicine, including education, leading to a broader approach to medical practice than the purely scientific approach. This is likely to enrich the profession and individuals therein. As well giving as a wider general education, areas of medical training and practice that a literary education will benefit directly include critical reading and appraisal, communication skills, history taking, 'surrogate experience', understanding the role of the physician, ethics, and self-expression. Many of these are central to our understanding of good medical practice. PMID- 9747555 TI - Acute sinusitis and antibiotic treatment. PMID- 9747556 TI - Urine sample collection. PMID- 9747557 TI - Community hospitals. PMID- 9747558 TI - Telling the truth. PMID- 9747559 TI - Prevalence and treatment of dizziness. PMID- 9747560 TI - Group D streptococcal throat infection. PMID- 9747561 TI - Screening: the inadequacy of population registers. PMID- 9747562 TI - Screening for abdominal aortic aneurysms in general practice. PMID- 9747563 TI - Developments in cardiovascular ultrasound. Part 2: Arterial applications. AB - Many of the changes resulting from arterial disease can be measured, using Doppler ultrasound for measurement of blood velocity and B-scan imaging for measurement of tissue structure and composition. Wall thickness, the degree of arterial narrowing and plaque volume can be measured using B-scan imaging, and 3D ultrasound can be used to improve the accuracy of measurements of plaque volume and for improved visualisation of complex arterial geometries. Measurement of the dynamic properties of the arterial wall permits estimation of wall elasticity and plaque motion. From the Doppler signal, measurements of blood velocity are used to estimate the degree of arterial narrowing and volumetric flow, although measurement errors can be large. Wall shear stress can be estimated by measuring the velocity gradient at the vessel wall. The problems of inadequate spatial resolution and interference from overlying tissue are largely removed when intravascular systems are used, and these have superior capability in the assessment of arterial structure and tissue composition. However, measurement of quantities relating to blood flow is more difficult using the intravascular approach, as the indwelling cather disturbs the blood flow pattern, and currently, assessment of flow and vessel cross-section are not performed at the same site. PMID- 9747564 TI - Gabor spectra for Doppler echocardiography. AB - Doppler echocardiography on current ultrasonography machines makes use of short time Fourier spectrograms (STFTs) to display the patterns of blood velocities in the heart chamber. From these sonologists infer the functional parameters, such as valve area, that have pathological significance. After mention of the lacuna in such an STFT display, the Gabor transform and associated Gabor spectrogram evaluation techniques are described. Finally, comparative views of ultrasound machine display with PC-based STFT and Gabor displays for two typical patients are shown, with inferences in support of the proposed Gabor display. PMID- 9747565 TI - Imaging and modelling from serial microscopic sections for the study of anatomy. AB - A system is considered for segmenting noisy intensity images and consequent three dimensional object reconstruction from a set of planar contours. A new semi automatic method for the extraction of contours from a sequence of cross sectional images based on an active contour model (ACM) is proposed. The dynamic ACM proceeds along the sequence of cross-sections following a non-rigid motion, in accordance with the organ boundary. Image texture information is also employed in the model. Problems associated with topological reconstruction from planar contours are addressed, and several criteria promoting semi-automatic topological reconstruction are introduced. The proposed system is successfully applied to the processing of real data related to animal embryonic organs, proving that the system allows detailed modelling of irregular objects. The reconstructed models can be observed in wire-frame, solid, transparent or stereoscopic semi transparent format. The human-computer interaction implemented in the procedure assists with problems of feature identification and object manipulation about an arbitrary axis. PMID- 9747566 TI - Non-radiological technique for 3D imaging of intestinal endoscopes: computerised graphical 3D representation of endoscope and skeleton. AB - Colorectal cancer is a common malignancy but as yet there is no agreement regarding the optimal method for screening. Colonoscopy is theoretically the investigation of choice. The examination can, however, be difficult to perform and the average trainee requires at least 200 supervised examinations to become proficient. Colonoscopy takes on average about half an hour per patient and sedation is normally required because of painful instrument looping. The authors previously developed a non-radiological method of visualising the path of the endoscope using magnetic drive coils under the patient and a chain of sensors in the biopsy channel of the instrument. The computer-generated grey-scale images produced in real time were deemed unsatisfactory and the anatomical markers confusing. A new computer graphics system is described in which a much more realistic endoscope and, if necessary, skeleton can be produced. The wire-frame octagonal representation should help in the detailed analysis of colonoscopy using existing endoscopes and aid in future computer design and testing of novel instruments incorporating worm or snake-like properties. PMID- 9747567 TI - Semi-automatic tool for segmentation and volumetric analysis of medical images. AB - Segmentation software is described, developed for medical image processing and run on Windows. The software applies basic image processing techniques through a graphical user interface. For particular applications, such as brain lesion segmentation, the software enables the combination of different segmentation techniques to improve its efficiency. The program is applied for magnetic resonance imaging, computed tomography and optical images of cryosections. The software can be utilised in numerous applications, including pre-processing for three-dimensional presentations, volumetric analysis and construction of volume conductor models. PMID- 9747568 TI - Focusing and targeting of magnetic brain stimulation using multiple coils. AB - Neurones can be excited by an externally applied time-varying electromagnetic field. Focused magnetic brain stimulation is attained using multiple small coils instead of one large coil, the resultant induced electric field being a superposition of the fields from each coil. In multichannel magnetic brain stimulation, partial cancellation of fields from individual coils provides a significant improvement in the focusing of the stimulating field, and independent coil channels allow targeting of the stimuli on a given spot without moving the coils. The problem of shaping the stimulating field in multichannel stimulation is analysed, and a method is derived that yields the driving currents required to induce a field with a user-defined shape. The formulation makes use of lead fields and minimum-norm estimation from magneto-encephalography. Using these methods, some properties of multichannel coil arrays are examined. Computer assisted multichannel stimulation of the cortex will enable several new studies, including quick determination of the cortical regions, the stimulation of which disrupts cortical processing required by a task. PMID- 9747569 TI - Application of the matching pursuit method for structural decomposition and averaging of phonocardiographic signals. AB - The paper evaluates the performance of an automatic adaptive time-frequency method to detect each cardiac cycle of a phonocardiogram (PCG) and extract average heart sounds and PCG cycles. The proposed method combines a global search of the PCG, in terms of the energy distribution of the most important components, with a local search relating to the specific events found within a cardiac cycle. The method is applied to 100 PCG recordings from 50 patients with an aortic bioprosthetic valve. The performance of the proposed method is compared with a commonly used semi-automatic method that is based on the combined analysis of an electrocardiogram (ECG) and the PCG signal. Results show that the proposed method clearly outperforms the semi-automatic method, especially in the case of patients with malfunctioning bioprostheses. By eliminating the need to record an ECG as the time-reference signal, this method reduces hardware overheads when analysis of PCG signals is the primary aim. It is also independent of subjective human judgment for selection of reference templates and threshold levels. Furthermore, the method is robust to artefacts, background noise and other kinds of signal interferences. With minor modifications, the procedure described could be applied to other types of biomedical signal in order to extract coherent transient components and identify specific events. PMID- 9747570 TI - Feature extraction for on-line EEG classification using principal components and linear discriminants. AB - The study focuses on the problems of dimensionality reduction by means of principal component analysis (PCA) in the context of single-trial EEG data classification (i.e. discriminating between imagined left- and right-hand movement). The principal components with the highest variance, however, do not necessarily carry the greatest information to enable a discrimination between classes. An EEG data set is presented where principal components with high variance cannot be used for discrimination. In addition, a method based on linear discriminant analysis (LDA), is introduced that detects principal components which can be used for discrimination, leading to data sets of reduced dimensionality but similar classification accuracy. PMID- 9747571 TI - Frequency-domain analysis of cerebral autoregulation from spontaneous fluctuations in arterial blood pressure. AB - The dynamic relationship between spontaneous fluctuations of arterial blood pressure (ABP) and corresponding changes in cerebral blood flow velocity (CBFV) is studied in a population of 83 neonates. Static and dynamic methods are used to identify two subgroups showing either normal (group A, n = 23) or impaired (group B, n = 21) cerebral autoregulation. An FFT algorithm is used to estimate the coherence and transfer function between CBFV and ABP. The significance of the linear dependence between these two variables is demonstrated by mean values of squared coherence > 0.50 for both groups in the frequency range 0.02-0.50 Hz. However, group A has significantly smaller coherences than group B in the frequency ranges 0.02-0.10 Hz and 0.33-0.49 Hz. The phase response of group A is also significantly more positive than that of group B, with slopes of 9.3 +/- 1.05, and 1.80 +/- 1.2 rad Hz-1, respectively. The amplitude frequency response is also significantly smaller for group A in relation to group B for the frequency range 0.25-0.43 Hz. These results suggest that transfer function analysis may be able to identify different components of cerebral autoregulation and also provide a deeper understanding of recent findings by other investigators. PMID- 9747572 TI - Accuracy of single-dipole inverse solution when localising ventricular pre excitation sites: simulation study. AB - Different factors are investigated that may affect the accuracy of an inverse solution that uses a single-dipole equivalent generator, in a standardised inhomogeneous torso model, when localising the pre-excitation sites. An anatomical model of the human ventricular myocardium is used to simulate body surface potential maps (BSPMs) and magnetic field maps (MFMs) for 35 pre excitation sites positioned on the epicardial surface along the atrioventricular ring. The sites of pre-excitation activity are estimated by the single-dipole method, and the measure for the accuracy of the localisation is the localisation error, defined as the distance between the location of the best-fitting single dipole and the actual site of pre-excitation in the ventricular model. The findings indicate that, when the electrical properties of the volume conductor and lead positions are precisely known and the 'measurement' noise is added to the simulated BSPMs and MFMs, the single-dipole method optimally localises the pre-excitation activity 20 ms after the onset of pre-excitation, within 0.71 +/- 0.28 cm and 0.65 +/- 0.30 cm using BSPMs and MFMs, respectively. When the standard torso model is used to localise the sites of onset of the pre-excitation sequence initiated in four individualised torso models, the maximum errors are as high as 2.6-3.0 cm (even though the average error, for both the BSPM and MFM localisations, remains within the 1.0-1.5 cm range). In spite of these shortcomings, it is thought that single-dipole localisations can be useful for non-invasive pre-interventional planning. PMID- 9747573 TI - Telemetry system for monitoring anterior cruciate ligament graft forces in vivo. AB - Quantifying changes in the tension of an anterior cruciate ligament (ACL) graft in vivo during rehabilitative exercises is useful for developing the optimum rehabilitation for patients who have had reconstructive surgery. The purpose of the work reported is to design, build and test a telemetry system that can measure the in vivo ACL graft tension post-operatively. A commercially available fixation device is modified to sense the graft tension, house electronic components, transmit an output signal and pass the power generating signal. A transcutaneous inductive link is used to power the implanted telemetry electronics. The current difference technique is used to measure changes in two resistance strain gauges that monitor shear strain developed on the femoral fixation device by the ACL graft. This current regulates a frequency-modulated output signal that is transmitted using a new technique. Harnessing the ionic and volume conduction properties of the body fluids, the new technique involves injecting current subcutaneously into the tissue and then sensing the potential developed on the skin by surface electrodes. The waveform shape, amount of charge injected, charge density and current density are regulated to avoid tissue damage, pain and unwanted muscular stimulation. A signal conditioning board detects and converts the output to an analogue voltage for collection by a computer data-acquisition system. A performance evaluation demonstrates that the telemetry system either meets or exceeds all of the criteria necessary for the application. PMID- 9747574 TI - Determination of total body water by multifrequency bio-electric impedance: development of several models. AB - Multifrequency bio-electronic impedance analysis (MF BIA) measurements are taken from a heterogeneous group of patients, varying in size between obese and slim. The measuring system uses four electrodes: two current and two potential electrodes. Three new models are developed to calculate total body water (TBW) from the BIA data, and the resulting TBW values are compared with TBW determined by D2O dilution. The results demonstrate that the most simple model provides the best TBW values. For individual patients, TBW can be determined by means of bioimpedance measurement with an accuracy of 3 litres. In the most simple model (model 1), the body is electrically represented by a cylinder, and corrections are made for the amount of fat. This is an extension of the model used by Xitron. In the more advanced models (2 and 3), the body is represented by a cylinder for the trunk, and truncated cones represent the arms and legs. In model 2, delta TBW amounts to 3 litres. It is shown that the resistance of the trunk is proportional to the square root of the length. In model 3, it is assumed that subcutaneous fat is a poor conductor of electric current. An equation is developed that describes the partition of subcutaneous fat, and the fat layer is then removed from the cones representing arms and legs and from the cylinder that models the trunk. PMID- 9747575 TI - Automated neural network detection of wavelet preprocessed electrocardiogram late potentials. AB - The aim of the study is to investigate the potential of a feedforward neural network for detecting wavelet preprocessed late potentials. The terminal parts of a simulated QRS complex are processed with a continuous wavelet transform, which leads to a time-frequency representation of the QRS complex. Then, diagnostic feature vectors are obtained by subdividing the representations into several regions and by processing the sum of the decomposition coefficients belonging to each region. The neural network is trained with these feature vectors. Simulated ECGs with varying signal-to-noise ratios are used to train and test the classifier. Results show that correct classification ranges from 79% (high-level noise) to 99% (no noise). The study shows the potential of neural networks for the classification of late potentials that have been preprocessed by a wavelet transform. However, clinical use of this method still requires further investigation. PMID- 9747576 TI - Electrostatic charge characteristics of Der p1 allergen-carrying particles and the house dust mite dermatophagoides pteronyssinus. AB - Control of the house dust mite allergen has received considerable attention owing to its importance in some allergic diseases. One aspect of dust mites and their allergen-carrying faecal particles that has not been reported on, which may have allergen control applications, is the electrostatic charge they carry in the natural environment. To promote tribo-electric charging, household dust containing dust mite allergen and live house dust mites are separately agitated while in contact with either polypropylene, nylon or earthed metal. The charged dust and mites are subsequently subjected to electrostatic separation and collection. Results for concentrations of the house dust mite allergen, Der p1, indicate that, when subjected to nylon, Der p1 carrier particles appear to be predominantly positively charged. Similarly, when subjected to polypropylene, Der p1 carrier particles also appear to be positively charged. Reduction of excess free charge by agitation against earthed metal does not appear to affect the observed charging characteristics, indicating that the positive charge may be bound or inherent in the Der p1 carrier particles. In contrast, house dust mites exposed to nylon appear to be generally charging negative, whereas mites exposed to polypropylene appear to be charging positive. The observed electrostatic characteristics of the mites and Der p1 carrying particles will be useful in the future development of electrostatic allergen control methods. PMID- 9747577 TI - Computer analysis system of blood oxygen saturation in an animal hypoxia model. AB - An experimental animal hypoxia model has been developed. It consists of two sensors (an in vitro and in vivo model), an experimental device and a computer signal processing system. This method can easily be applied to determine and analyse blood oxygen saturation at various hypoxia levels. It can also be used to evaluate the accuracy of pulse oximetry over a wide range of oxyhemoglobin desaturation levels. The DC and AC components of recorded red and infra-red signals, the dual-wavelength ratio R12 and the reading of a pulse oximeter (SpO2) can be automatically calculated and displayed on a computer screen. Preliminary results of the animal hypoxia test indicate that the measurements made by the instrument correlate well with the oxygen saturation readings of the automatic blood gas analyser AVL945. The computer analysis system is suitable for repeated estimations in the animal model. PMID- 9747579 TI - On-line control of cellular adhesion with impedance measurements using interdigitated electrode structures. AB - Critical parameters to be assessed in cell culture are the number of viable cells and cell viability. Growth, product formation, toxicity effects and the overall success of cell culture can depend largely on these. With interdigitated electrode structures (IDES) adherent cells are cultured directly on a pair of interdigitated electrodes, and the impedance of the system gives insight into the adhesive behaviour of the cells. The signal is influenced by the changes in number, growth and morphological behaviour of adherently growing cells, mainly owing to the insulating effects of the cell membranes. Five different cell lines are used, and their divergent behaviour is monitored over a period of four days, from inoculation of the cells to killing of the cells at the end of the experiments. Even when the cells from close monolayers, great fluctuations in the impedance signal can be observed. Nevertheless, for a more complete description of cellular systems, other parameters, such as acidification and respiration, have to be included in the measuring system. PMID- 9747578 TI - Engineering the healing of the rabbit medial collateral ligament. AB - A biological approach to improve healing of the medial collateral ligament (MCL) was investigated by exploring the use of therapeutic growth factors based on in vitro and in vivo experiments. The in vitro cell culture studies involved screening a variety of growth factors to select those that exhibit the most positive effects on cell proliferation and extracellular matrix synthesis. The selected growth factors were applied in vivo to a rabbit model where the MCL was ruptured. Biomechanical and histological evaluations are performed to determine whether the selected growth factors can enhance the properties of the healed MCL, whether these improvements are dose dependent, and whether combinations of growth factors can enhance MCL healing to a greater extent than individual growth factors. In vitro studies showed that epidermal growth factor (EGF) and platelet derived growth factor-BB (PDGF-BB) have the greatest effect on ligament fibroblast proliferation, whereas transforming growth factor-beta 1 (TGF-beta 1) superiorly promotes extracellular matrix synthesis. These growth factors were then applied in vivo at different dosages, in isolation and in combination, and the ligaments were evaluated six weeks post-operatively. Tensile testing of the femur-MCL-tibia complexes (FMTCs) revealed that the specimens treated with a high dose of PDGF-BB have ultimate load, ultimate elongation and energy absorbed to failure values that are significantly greater than those from the other groups. The high dose of PDGF-BB was more effective than the low dose, indicating a dose dependency. The addition of TGF-beta 1 to PDGF-BB did not lead to any further increases in the structural properties of the FMTC. These encouraging results suggest that PDGF-BB may be a potential growth factor to enhance the quality of the healing ligament. PMID- 9747581 TI - A review of sunscreen safety and efficacy. AB - The use of sunscreen products has been advocated by many health care practitioners as a means to reduce skin damage produced by ultraviolet radiation (UVR) from sunlight. There is a need to better understand the efficacy and safety of sunscreen products given this ongoing campaign encouraging their use. The approach used to establish sunscreen efficacy, sun protection factor (SPF), is a useful assessment of primarily UVB (290-320 nm) filters. The SPF test, however, does not adequately assess the complete photoprotective profile of sunscreens specifically against long wavelength UVAI (340-400 nm). Moreover, to date, there is no singular, agreed upon method for evaluating UVA efficacy despite the immediate and seemingly urgent consumer need to develop sunscreen products that provide broad-spectrum UVB and UVA photoprotection. With regard to the safety of UVB and UVA filters, the current list of commonly used organic and inorganic sunscreens has favorable toxicological profiles based on acute, subchronic and chronic animal or human studies. Further, in most studies, sunscreens have been shown to prevent the damaging effects of UVR exposure. Thus, based on this review of currently available data, it is concluded that sunscreen ingredients or products do not pose a human health concern. Further, the regular use of appropriate broad-spectrum sunscreen products could have a significant and favorable impact on public health as part of an overall strategy to reduce UVR exposure. PMID- 9747580 TI - Regulation of neurofilament interactions in vitro by natural and synthetic polypeptides sharing Lys-Ser-Pro sequences with the heavy neurofilament subunit NF-H: neurofilament crossbridging by antiparallel sidearm overlapping. AB - Neurofilaments are organised into parallel bundles in axons through crossbridges formed by lateral projections of neurofilament subunits. Pure neurofilaments form gels in vitro, consisting of interconnected parallel arrays of filaments regulated by the phosphorylation level of neurofilament subunits. Neurofilament associated polypeptides sharing phosphorylated epitopes with the repetitive lysine-serine-proline (Lys-Ser-Pro) motifs of the neurofilament heavy subunit sidearm are characterised: they regulate in vitro the neurofilament gelation kinetics in a concentration- and phosphorylation-dependent manner. Studies with synthetic peptides show that interactions between neurofilaments involve both acid and base amino acid residues of neurofilament sidearms and demonstrate the opposite effects of peptides containing either one (inhibition) or two (activation) Lys-Ser-Pro motifs. Electron microscopy reveals an organised network of native neurofilament sidearms, regulated by the phosphorylation level of neurofilament subunits, suggesting a structural transition between intra- and inter-neurofilament sidearm interactions. These results favour the hypothesis of a mechanism of neurofilament crossbridging through the variable antiparallel overlapping of the phosphorylable Lys-Ser-Pro domains of neurofilament sidearms from adjacent filaments, following an equilibrium regulated by neurofilament associated proteins, bivalent cations and the phosphorylation level of Lys-Ser Pro motifs from both neurofilament sidearms and neurofilament-associated proteins. PMID- 9747582 TI - Exclusive free radical mechanisms of cellular photosensitization. AB - In order to determine the specific effects of radical-induced reactions in the absence of complicating excited-state pathways, four different thiohydroxamic esters and their parent molecule, N-hydroxypyridine-2(1H)-thione, have been studied in murine L1210 leukemia cells for their ability to produce photobiological damage. Irradiation (lambda exc = 355 nm) of cells in the presence of thiopyridone esters, specific photolytic precursors of sulfur-, carbon- and oxygen-centered radicals, caused toxicity that was unambiguously demonstrated to result from radical photosensitization mechanisms. Cellular morphological changes were observed following irradiation but apoptosis was not found to take place. A good correlation was evident between lipid peroxidation, measured by the thiobarbituric acid method, and phototoxicity, assessed by the trypan blue exclusion assay, indicating that the ester derivatives exert their effects mainly in plasma and/or subcellular membranes. Irradiation performed under deaerated conditions also induced significant phototoxicity but the effects of deaeration were dependent on the ester used and are discussed in terms of the nature of the primary radical species generated in each case. Irradiation of L1210 cells in the presence of N-hydroxypyridine-2(1H)-thione, a nonspecific, photochemical source of hydroxyl radical, was also found to trigger phototoxicity and lipid peroxidation. However in this case, photodamage cannot yet be definitely attributed to a radical or type II mechanism although the apparent oxygen independence of phototoxicity would indicate that type II contribution is not significant. PMID- 9747583 TI - Light dosimetry for intraperitoneal photodynamic therapy in a murine xenograft model of human epithelial ovarian carcinoma. AB - Few studies have been published to date measuring spatially resolved fluence rates in complex tissue geometries. Here the light distributions of three different intraperitoneal light delivery geometries in a murine ovarian cancer model were investigated to assess their influence on the tumorcidal efficacy of photodynamic therapy (PDT). In vivo fluence rate measurements in the peritoneal cavities of mice, with the light intensity being mapped in three transverse planes, were performed using fiber-optic detectors. Three different source fiber designs and placements were tested for their ability to provide uniform irradiation of the peritoneal cavity. The biological response to a PDT protocol comprising three separate treatments administered at 72 h intervals, each consisting of a 0.25 mg kg-1 intraperitoneal injection of benzoporphyrin derivative-mono acid ring A followed 90 min later by delivery of 15 J of 690 nm light, was measured. The tissue response was evaluated by measuring the number of remaining visible lesions and the total residual tumor mass. Fluence rate measurements showed large variations in the fluence rate distribution for similar intended treatments. The most uniform and reproducible illumination was achieved using two 18 mm long cylindrical emitting optical fibers. The biological response was comparable to that produced when a flat-cleaved end optical fiber is used to illuminate the four quadrants of the abdomen sequentially. While a good reproducibility in tumor induction in this animal model exists, no correlation was found between the fluence rate distribution measured in one group of animals and the biological response in a separate group of similarly treated animals. Due to the large intra-animal variability in fluence rate distribution, representative fluence rate mapping in complex tissue geometries is of limited value when applied to an individual PDT treatment. Thus, surveillance of the fluence rate during individual treatments will be required for acceptable PDT dosimetry. To improve the versatility of this particular animal model for PDT research, a large number of extended sources are required to increase uniformity of the illumination in order to reduce unwanted cytotoxic side effects resulting from foci of high fluence rates. In this way, subsequent increase of the total energy delivered to the tumor may be possible. PMID- 9747585 TI - Wavelength-dependent photoacoustic calorimetry study of melanin. AB - Photoacoustic calorimetry is used to examine the energy dissipation in melanin under physiological conditions (pH 7.2) following irradiation by UV and visible (VIS) light. Four different excitation wavelengths were examined: 264 nm, representative of UVC radiation, 351 nm and 400 nm (UVA-I radiation) and 527 nm, representative of VIS radiation. Following absorption at 527 nm, essentially all of the photon energy is released nonradiatively on a sub-nanosecond of excitation. Similar results are observed at 400 nm. At 351 nm, most of the energy was released as heat; a small amount of energy was retained (5 +/- 5%). When melanin is excited at 264 nm, 29 +/- 7% of the photon energy is retained by the molecule for a time period longer than a few hundred nanoseconds. These results show that a long-lived excited state or reactive intermediate is generated upon UV irradiation, whereas all of the excitation energy is dissipated nonradiatively in the visible portion of the spectrum. These results establish that the photochemistry of melanin is wavelength dependent. PMID- 9747584 TI - DNA photodamage, repair, gene induction and genotoxicity following exposures to 254 nm UV and 8-methoxypsoralen plus UVA in a eukaryotic cell system. AB - The induction and repair of different types of photodamage and photogenotoxicity in eukaryotic cells have been the subject of many studies. Little is known about possible links between these phenomena and the induction of DNA damage-inducible genes. We explored this relationship using the yeast Saccharomyces cerevisiae, a pertinent eukaryotic model. Previous results showed that the photogenotoxic potential of 8-methoxypsoralen (8-MOP) plus UVA is higher than that of UV (254 nm). Moreover, the induction of the ribonucleotide reductase gene RNR2 by UV and 8-MOP plus UVA in an RNR2-LACZ fusion strain and the formation of DNA double strand breaks (dsb) as repair intermediates after such treatments suggest that the latter process could involve a signal for gene induction. To further substantiate this, we measured the induction of the DNA repair gene RAD51 in RAD51-LACZ fusion strains using the dsb repair and recombination deficient mutant rad52 and the corresponding wild type, and we determined the formation of dsb by pulsed-field gel electrophoresis. After treatments, the resealing of dsb formed as repair intermediates was impaired in the rad52 mutant. At equal doses, i.e. the same number of lesions, the induction of the RAD51 gene by UV or 8-MOP plus UVA was significantly reduced in the rad52 mutant as compared with the wild type. The same was true when equitoxic doses were used. Thus, the RAD52 repair pathway appears to play an important role not only in dsb repair but also in gene induction. Furthermore, the signaling pathways initiated by DNA damage and its processing are somewhat linked to the photogenotoxic response. PMID- 9747586 TI - Probing the binding region of the single-stranded DNA-binding domain of rat DNA polymerase beta using nanosecond-pulse laser-induced cross-linking and mass spectrometry. AB - In recent years, there has been a significant number of studies in which UV light has been used as a reagent to induce cross-links in nucleic acid-protein complexes. An area of considerable interest among those interested in structural biology is the garnering of information about the sites of cross-linking within the protein and nucleic acid members of photolinked conjugates, under the assumption that such knowledge should lead to identification of contact regions or sites within the native complexes. In this paper, we present our results from a photocross-linking study of the complex of the single-stranded DNA-binding domain of rat DNA polymerase beta (pol beta-ss) with the oligonucleotide d(ATATATA). In this study, we have used single nanosecond laser pulses as the cross-linking reagent and matrix-assisted laser desorption/ionization-time of flight mass spectrometry as an analytical tool to identify cross-linked peptides purified from proteolytic digests of the cross-linked complex. Six cross-linked peptides have been identified in tryptic digests of the protein-oligonucleotide conjugates that result from irradiation of the pol beta-ss-d(ATATATA) complex with a single laser pulse. Comparisons with NMR data in the literature for the same complex show that each of the cross-linked peptides contains amino acids that are in contact with the nucleic acid component of the complex. PMID- 9747587 TI - Ultraviolet-A-dependent inhibition of cytoplasmic aconitase activity of iron regulatory protein-1 in NCTC 2544 keratinocytes. AB - The aconitase activity of the cytoplasmic iron regulatory protein-1 of NCTC 2544 keratinocytes is effectively inhibited by physiological doses of UVA. The time course of the photoinactivation is biphasic. A fast step is first observed corresponding to about 50% inactivation after exposure to 5 J/cm2 of UVA followed by a much slower photoinactivation at higher doses. The water-soluble antioxidant N-acetylcysteine only partially inhibits the photoinduced inactivation of the cytoplasmic aconitase function, whereas the lipophilic vitamin E, the iron chelator, desferrioxamine and the superoxide dismutase inhibitor, diethyldithiocarbamate do not protect at all. As a consequence, reactive oxygen species such as O2-., H2O2 and lipid peroxides and hydroperoxides seem to play a rather minor role in the inactivation induced by the UVA photooxidative stress although an oxidative stress produced by O2-. and H2O2 is known to inhibit reversibly and effectively cytoplasmic aconitase activity in mammalian cells. PMID- 9747588 TI - Release of inflammatory mediators (PGE2, IL-6) by fenofibric acid-photosensitized human keratinocytes and fibroblasts. AB - Ultraviolet-A radiation has weak effects on the release of inflammatory mediators by skin cells due to the poor overlap between UVA wavelengths and the absorption spectra of the relevant chromophores of key biomolecules. However, this situation could be very different in the presence of a photosensitizing drug. To investigate this issue, we have irradiated human skin cells (keratinocytes and fibroblasts) in the presence of fenofibric acid (the active phototoxic metabolite of fenofibrate). The results of this research show a dual effect on the production/release of inflammatory mediators: the synthesis of the proinflammatory cytokine interleukin-6 becomes strongly inhibited at photosensitizer concentrations that clearly stimulate the production of prostaglandins (PGE2) by skin cells. We have found evidences showing that the de novo synthesis of cytokines is inhibited in photosensitized cells due to the fact that cellular mRNA is degraded. Interestingly, when the medium taken from irradiated cultures is added to nonexposed cells, a significant stimulation of cytokine synthesis is observed that can be inhibited by anti-PGE2 antibodies. These observations may be relevant in vivo, where prostaglandins released by photosensitized skin cells could stimulate cytokine synthesis by underlying, nonirradiated cells. PMID- 9747589 TI - Photomodification of cardiac membrane: chaotic currents and high conductance states in isolated patches. AB - We have demonstrated previously that photomodification permeabilizes cardiac cells as evidenced by activation of a whole-cell leak current. In this paper we report that photomodification induces in cell-attached and inside-out cardiac membrane patches a chaotic current. Unlike current recordings from many protein ion channels that show stepwise amplitude changes associated with open and closed states of the channel, the chaotic current consists of variable amplitude spike like transitions. The amplitudes of these spikes can vary from tenths to tens of picoamperes at a constant transmembrane potential. We provide evidence that the chaotic current is transmembrane rather than trans-seal and has a voltage dependency expected for current flow through nonspecific conductance pathways. Photomodification can also induce high conductance states (greater than 500 pS) in cell-attached and inside-out patches. We present evidence that the high conductance state is also not related to seal breakdown. Our results suggest that both the chaotic current activity in and high conductance state of photomodified cardiac membrane patches result from the opening of many small conductance, nonspecific pathways through the membrane. PMID- 9747590 TI - Stationary headband for clinical time-of-flight optical imaging at the bedside. AB - Conventional brain-imaging modalities may be limited by high cost, difficulty of bedside use, noncontinuous operation, invasiveness or an inability to obtain measurements of tissue function, such as oxygenation during stroke. Our goal was to develop a bedside clinical device able to generate continuous, noninvasive, tomographic images of the brain using low-power nonionizing optical radiation. We modified an existing stage-based time-of-flight optical tomography system to allow imaging of patients under clinical conditions. First, a stationary head band consisting of thin, flexible optical fibers was constructed. The headband was then calibrated and tested, including an assessment of fiber lengths, the existing system software was modified to collect headband data and to perform simultaneous collection of data and image reconstruction, and the existing hardware was modified to scan optically using this headband. The headband was tested on resin models and allowed for the generation of tomographic images in vitro; the headband was tested on critically ill infants and allowed for optical tomographic images of the neonatal brain to be obtained in vivo. PMID- 9747591 TI - A study of the uptake of toluidine blue O by Porphyromonas gingivalis and the mechanism of lethal photosensitization. AB - The purpose of the study was to determine the distribution of the photosensitizer toluidine blue O (TBO) within Porphyromonas gingivalis and the possible mechanism(s) involved in the lethal photosensitization of this organism. The distribution of TBO was determined by incubating P. gingivalis with tritiated TBO (3H-TBO) and fractionating the cells into outer membrane (OM), plasma membrane (PM), cytoplasmic proteins, other cytoplasmic constituents and DNA. The percentage of TBO in each of the fractions was found to be, 86.7, 5.4, 1.9, 5.7 and 0.3%, respectively. The involvement of cytotoxic species in the lethal photosensitization induced by light from a heliumneon (HeNe) laser and TBO was investigated by using deuterium oxide (D2O), which prolongs the lifetime of singlet oxygen, and the free radical and signlet oxygen scavenger L-tryptophan. There were 9.0 log10 and 2 log10 reductions in the presence of D2O and H2O (saline solutions), respectively, at a light dose of 0.44 J (energy density = 0.22 J/cm2), suggesting the involvement of singlet oxygen. Decreased kills were attained in the presence of increasing concentrations of L-tryptophan. The effect of lethal photosensitization on whole cell proteins was determined by measuring tryptophan fluorescence, which decreased by 30% using 4.3 J (energy density = 4.3 J/cm2) of light. Effects on the OM and PM proteins were determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. There was evidence of change in the molecular masses of several PM proteins and OM proteins compared to controls. There was evidence of damage to the DNA obtained from irradiated cells. Scanning electron microscopic studies showed that there was coaggregation of P. gingivalis cells when sensitized and then exposed to laser light. These results suggest that lethal photosensitization of P. gingivalis may involve changes in OM and/or PM proteins and DNA damage mediated by singlet oxygen. PMID- 9747592 TI - Treatment of malignant melanoma by high-peak-power 1064 nm irradiation followed by photodynamic therapy. AB - Irradiation of B16 pigmented melanoma subcutaneously transplanted in C57 mice with a single 650 mJ pulse (10 ns) of 1064 nm light from a Q-switched Nd: YAG laser caused instantaneous bleaching of the pigmented tissue. Visual and histological examination of the resulting gray-colored tumor revealed the breakdown of melanosomes with no detectable alteration of the normal and tumor overlying skin. Histological examination of the irradiated tumor showed some degree of vascular damage; the depth of the photodamage was not affected by the successive delivery of three consecutive light pulses. The bleached tumor grew at a modestly slower rate but the high-peak-power (HPP) laser treatment did not affect the tumor concentration of a photodynamic sensitizer Si(i.v.) naphthalocyanine (isoBO-SiNc) intravenously injected 24 h before Nd:YAG irradiation. Treatment of the B16 pigmented melanoma by photodynamic therapy (PDT: 1 mg/kg isoBO-SiNc, 300 mW/cm2, 520 J/cm2) from a 774 nm diode laser immediately after the 1064 nm irradiation resulted in a 16 day delay of tumor regrowth, which was markedly longer than the delay (ca 6 days) obtained after PDT under identical conditions without the preirradiation. Thus, pretreatment of pigmented tumors with HPP 1064 nm light appears to enhance their susceptibility to conventional PDT. The tumor response was further enhanced by repeating the combined HPP/PDT treatment at an interval of 10 days (regrowth delay: 27 days), as well as by applying hyperthermia immediately after HPP/PDT (regrowth delay: ca 34 days). PMID- 9747593 TI - Clinical evaluation of the cutaneous phototoxicity of 5,10,15,20-tetra(m hydroxyphenyl)chlorin. AB - The cutaneous phototoxic reaction induced by intravenous injection of 5,-10,-15, 20-tetra(m-hydroxyphenyl)chlorin (mTHPC) has been clinically evaluated in patients undergoing photodynamic therapy. These tests were performed on the backs of 23 patients with a solar simulator at various times after drug administration ranging from 5 h to 57 days. The mTHPC doses ranged from 0.1 to 0.3 mg/kg, and the illuminations lasted from 30 s up to 8 min. These tests have shown that the duration of the skin photosensitization induced after a typical therapeutic dose of mTHPC (0.15 mg/kg) is less important than with Photofrin (2 mg/kg). The level of mTHPC in the skin was also assessed in vivo and at times corresponding to the irradiations using an optical fiber-based spectrofluorometer. This study indicates that the light-induced fluorescence spectroscopy of mTHPC enables prediction of the degree of photosensitivity of the skin. PMID- 9747594 TI - Possible mechanisms of vascular relaxation induced by pulsed-UV laser. AB - This study was designed to examine the mechanism of vasorelaxation induced by pulsed-UV laser. Luminal diameters of rat femoral arteries were measured prior to and following krypton-fluoride excimer laser irradiation of 248 nm in wavelength. The diameter was enlarged to 1.3 times the preirradiated size at 1 or 10 Hz irradiation when the fluence was over 2.0 mJ/pulse/mm2, while the diameter reached 1.8 times at 100 Hz with a fluence of 0.8 mJ/pulse/mm2. Vasorelaxation by the 100 Hz irradiation was inhibited when the artery was pretreated with methylene blue but was enhanced with superoxide dismutase. Pathological analysis revealed an ablation crater and vacuole formation in the vessel at 1 or 10 Hz irradiation, but these changes were not remarkable in the 100 Hz-exposed sample. These findings suggest that vasorelaxation induced by the pulsed UV irradiation at 1 or 10 Hz results from structural alteration of vascular smooth muscle by the ablation crater or vacuolization. On the other hand, a possible mechanism of vasorelaxation at the 100 Hz irradiation is partially related to nitric oxide. PMID- 9747595 TI - Comparison of the in vivo photodynamic threshold dose for photofrin, mono- and tetrasulfonated aluminum phthalocyanine using a rat liver model. AB - The photodynamic threshold dose in normal rat liver was determined from the measured depth of necrosis following surface irradiation. The threshold was determined for the photosensitizing drugs Photofrin and monosulfonated aluminum chlorophthalocyanine, AlPcS1, at 24 h postinjection and was found to be (3.4 x/divided by 1.3) x 10(18) and (8.2 x/divided by 1.5) x 10(18) photons cm-3, respectively, compared with the previously reported value of (38 +/- 2) x 10(18) photons cm-3 for the tri/tetrasulfonated phthalocyanine, AlPcS4. These values were independent of drug concentration or total light fluence. For all three drugs the depth of tissue necrosis decreased as the time between drug and light administration increased from 10 min to 72 h. This decrease can be attributed both to the change in the tissue drug concentration as well as to changes in the efficiency of photodynamic therapy for producing tissue damage, related to the photodynamic necrosis threshold. The threshold values for all three photosensitizers were lowest at 10 min post injection: (1.4 x/divided by 1.4) x 10(18), (1.6 x/divided by 1.3) x 10(18) and (23 x/divided by 1.3) x 10(18) photons cm-3 for Photofrin, AlPcS1 and AlPcS4, respectively. The changes in necrosis threshold with time may be due to an initial change from entirely vascular to a combination of vascular and cellular damage, with later redistribution of the photosensitizer to targets at the subcellular level. PMID- 9747596 TI - Light irradiation of mouse spermatozoa: stimulation of in vitro fertilization and calcium signals. AB - Irradiation of mouse spermatozoa by 630 nm He-Ne laser was found to enhance the intracellular calcium levels and fertilizing potential of these cells. The effect of light on calcium transport and on fertilization rate was abrogated in the absence of Ca2+ during the irradiation time, indicating that the effect of light is Ca2+ dependent. The stimulatory effect of light on Ca2+ uptake was abolished in the presence of a voltage-dependent Ca(2+)-channel inhibitor nifedipine, indicating the involvement of a plasma membrane voltage-dependent Ca2+ channel. Furthermore, the stimulatory effect of light was completely inhibited by the mitochondrial uncoupler FCCP, indicating that laser irradiation might affect the mitochondrial Ca2+ transport mechanisms. A causal association between laser irradiation, reactive oxygen species (ROS) generation and sperm function was indicated by studies with ROS scavengers, superoxide dismutase (SOD) and catalase, and exogenous hydrogen peroxide. The SOD treatment, which enhanced H2O2 production, resulted in increased Ca2+ uptake and enhanced fertilization rate. On the other hand, catalase, which decomposes H2O2, impaired the light-induced stimulation in Ca2+ uptake and the fertilization rate. Taken together, the data suggest that H2O2 might be involved in the irradiation effects, and indeed laser irradiation enhances the production of H2O2 by spermatozoa. These results indicate that the effect of 630 nm He-Ne laser irradiation is mediated through the generation of H2O2 by the spermatozoa and that this effect plays a significant role in the augmentation of the sperm cells' capability to fertilize metaphase II-arrested eggs in vitro. PMID- 9747597 TI - Development of a fiber optic probe to measure NIR Raman spectra of cervical tissue in vivo. AB - The goal of this study was to develop a compact fiber optic probe to measure near infrared Raman spectra of human cervical tissue in vivo for the clinical diagnosis of cervical precancers. A Raman spectrometer and fiber optic probe were designed, constructed and tested. The probe was first tested using standards with known Raman spectra, and then the probe was used to acquire Raman spectra from normal and precancerous cervical tissue in vivo. Raman spectra of cervical tissue could be acquired in vivo in 90 s using incident powers comparable to the threshold limit values for laser exposure of the skin. Although some silica signal obscured tissue Raman bands below 900 cm-1, Raman features from cervical tissue could clearly be discerned with an acceptable signal-to-noise ratio above 900 cm-1. The success of the Raman probe described here indicates that near infrared Raman spectra can be measured in vivo from cervical tissues. Increasing the power of the excitation source could reduce the integration time to below 20 s. PMID- 9747598 TI - Interactive processes during interlimb coordination: combining movement patterns with different frequency ratios. AB - The present study examined the formation of a movement pattern that was added to an ongoing coordinative regime across different limb combinations. It was hypothesized that the addition of the secondary mode would perturb the ongoing primary mode by adding rhythmic complexity to the task requirements. Furthermore, the formation of the secondary mode was predicted to be affected by the ongoing coordination pattern. In Exp. 1, a primary multifrequency mode (2:1 ratio) was performed while a secondary isofrequency mode (1:1 ratio) was initiated midway into the trials, whereas the reversed dual-pattern conditions were examined in Exp. 2. The results from both experiments showed that the multifrequency mode deteriorated across limb combination under dual-pattern as compared to single pattern conditions. The isofrequency mode was also affected under combined pattern conditions, but its degradation was a function of the limb combination under consideration. In particular, the non-homologous limbs, which demonstrated less stable behavior than the homologous limbs under single-pattern conditions, were affected most strongly when confronted with the simultaneous production of the multifrequency mode. In addition, anti-phase movements deteriorated more than in-phase movements, supporting indirectly the contention that afferent feedback monitoring complexity differs for the two movement configurations. The findings of this study suggest that manipulation of task requirements can be used to examine pattern durability and formation in view of dynamical perturbations. PMID- 9747599 TI - Lexical access to inflected words as measured by lateralized visual lexical decision. AB - In two lateralized visual lexical decision experiments conducted with normal subjects, we studied hemispheric performance in the recognition of case-inflected Finnish nouns. Previous research employing mainly locative cases has indicated that such noun forms undergo morphological decomposition. The present experiments extend this finding to syntactic cases by showing that nouns with partitive or genitive endings take longer to recognize and elicit more errors than otherwise comparable monomorphemic nominative singular nouns. Morpheme-based recognition of all case-inflected forms would be a particularly appropriate solution for mental lexicon in highly inflecting languages like Finnish: it saves storage space and enables fast recognition of inflected forms not encountered before. In real words, morphological structure did not interact with visual field. However, particularly demanding, morphologically decomposable nonwords elicited more errors in the left visual field/right hemisphere than did nondecomposable nonwords. Our results suggest that at least in Finnish, both hemispheres are capable of morpheme-based access, but this mechanism is more accurate in the left than in the right hemisphere. PMID- 9747600 TI - Apoptosis in rat renal proximal tubular cells induced by cadmium. AB - Cadmium chloride can induce DNA fragmentation, a biochemical characteristic of apoptosis in renal epithelial LLC-PK, cells. This study was extended to determine the in vivo effects of this heavy metal on apoptosis. Nephrotoxicity was induced by a single intravenous administration of cadmium-metallothionein (0.15 mg metallothionein-bound cadmium/kg body weight) to male Jcl:Wistar rats. DNA fragmentation was seen in the kidney 12 h after injection of cadmium metallothionein without a concurrent release of lactate dehydrogenase in urine. Cycloheximide (3 mg/kg) inhibited cadmium-induced DNA fragmentation, suggesting that protein synthesis might be required for the induction of cell death by this metal. Apoptotic cells were identified in proximal tubular cells by in situ DNA 3'-end labeling. Furthermore, chromatin condensation in the apoptotic population of renal proximal tubular cells was noted. Data thus suggest that cadmium produces biochemical and morphological alterations in kidney, which are characteristic features seen in apoptosis. PMID- 9747601 TI - Basolateral uptake of mercuric conjugates of N-acetylcysteine and cysteine in the kidney involves the organic anion transport system. AB - Renal uptake and disposition of administered inorganic mercury were studied in rats that had undergone an acute bilateral ureteral ligation shortly before being injected intravenously with a nontoxic 0.5 micromol/kg dose of inorganic mercury with or without 2 micromol/kg N-acetylcysteine or cysteine. Bilateral ureteral ligation was performed in an attempt to reduce glomerular filtration to negligible levels, which in turn permitted the study of the basolateral uptake of inorganic mercury. The disposition of mercury was studied in the kidneys, liver, and blood 1 h after treatment. In rats given only mercuric chloride, the renal burden of mercury was approximately 20% of the administered dose of mercury. This confirms previous observations implicating a basolateral mechanism in the renal uptake of inorganic mercury. Coadministration of inorganic mercury with either N acetylcysteine or cysteine caused a significant increase in the renal uptake of mercury 1 h after treatment, particularly in the rats treated with inorganic mercury plus N-acetylcysteine. The enhanced uptake of mercury in the kidneys was due to increased uptake of mercury in the renal cortex and outer stripe of the outer medulla. Interestingly, the rate of uptake of mercury was so great in the rats treated with inorganic mercury plus N-acetylcysteine that the renal burden of mercury was virtually equivalent to that generally detected in normal animals administered the same dose of inorganic mercury as mercuric chloride. Pretreatment with para-aminohippuric acid (PAH) (which is a potent inhibitor of the organic anion transport system) caused significant reductions in the renal uptake and burden of inorganic mercury in all the rats administered inorganic mercury, regardless of whether the inorganic mercury was coadministered with N acetylcysteine or cysteine. Overall, the findings from the present study provide additional evidence that there is basolateral uptake of inorganic mercury in the kidneys, and that the primary or sole mechanism is dependent on the activity of the organic anion transporter. The present findings also show that cysteine and N acetylcysteine enhance the basolateral uptake of mercuric ions in the kidney when they are coadministered with inorganic mercury, presumably in the form of mercuric conjugates. Moreover, it appears that mercuric conjugates of N acetylcysteine are taken up more avidly at the basolateral membrane than mercuric conjugates of cysteine. Furthermore, the basolateral uptake of mercuric conjugates of N-acetylcysteine or cysteine in the kidney is due primarily to a mechanism involving the organic anion transport system. PMID- 9747602 TI - Environmental particulate-mediated cytokine production in lung epithelial cells (A549): role of preexisting inflammation and oxidant stress. AB - Epidemiologic data show that air pollution particulates cause adverse pulmonary health effects, especially in individuals with preexisting lung disease. We sought to model in vitro preexisting lung inflammation in order to investigate the hypothesis that "primed" lung epithelial cells will exhibit enhanced phlogistic responses [e.g., interleukin-8 (IL-8) production] to particulate air pollution. Exposure of tumor necrosis factor alpha (TNF-alpha) primed or control A549 cells to the air pollution particulates, residual oil fly ash (ROFA), and the known pathogenic dust alpha-quartz, but not inert TiO2, caused increased IL-8 production in primed cells compared to normal cells in a concentration-dependent manner (particle concentration range 0-200 microg/ml). We hypothesized that oxidant mechanisms may be involved in the cellular response to particulates. Addition of the antioxidant N-acetylcysteine (NAC, 1.0 mM) decreased ROFA and alpha-quartz-mediated IL-8 production by approximately 50% in normal and TNF alpha-primed A549 cells. In addition, exposure of A549 cells to ROFA caused a substantial (and NAC inhibitable) increase in oxidant levels as measured by fluorometry (DCFH oxidation). These data suggest that (1) lung epithelial cells primed by inflammatory mediators can show enhanced cytokine production after exposure to air pollution particulates, and (2) oxidant stress is a key mechanism for this response. PMID- 9747603 TI - Lead-induced changes in spermatozoa function and metabolism. AB - The relationships between sperm reactive oxygen species (ROS) generation, the capacitation process and acrosome reaction, and the spermoocyte penetration rate (SOPR) were investigated to understand the effect of lead toxicity on sperm functions and the mechanisms of these effects. Male Sprague-Dawley rats received weekly intraperitoneal injections of 20 mg or 50 mg lead acetate/kg or 20 mg or 50 mg sodium acetate/kg (control) for 6 wk. Serum testosterone was measured by radioimmunoassay. In cauda epididymal spermatozoa, the chemiluminescence was measured to evaluate the sperm ROS generation. Chlortetracycline fluorescence assay was used to study the status of capacitation and acrosome reaction on fresh cauda epididymal spermatozoa and after 2, 4, or 24 h of incubation with 5 mg/ml bovine serum albumin. In lead-exposed rats, the serum testosterone levels were reduced, and the percentage of capacitation and the chemiluminescence were significantly increased in fresh cauda epididymal spermatozoa. The serum testosterone levels were negatively associated with the percentage of acrosome reacted spermatozoa. Sperm chemiluminescence was positively correlated with the percentage of both capacitated and acrosome-reacted spermatozoa. The SOPR was negatively associated with the percentage of both capacitated and acrosome reacted spermatozoa. In summary, this study showed that male rats exposed to lead had decreased serum testosterone levels and that this metal produced early onset of capacitation by one of the pathways of ROS generation. These effects might consequently result in premature acrosome reaction and reduced zona-intact oocyte penetrating capability. PMID- 9747604 TI - NOAEL and LOAEL determinations of acute hepatotoxicity for chloroform and bromodichloromethane delivered in an aqueous vehicle to F344 rats. AB - Chloroform (CHCl3) and bromodichloromethane (BDCM) are generally the two most prevalent disinfection by-products formed during chlorination of drinking water, and both have been shown to be hepatotoxic, nephrotoxic, and carcinogenic in rodents. As the toxicity of these trihalomethanes (THMs) has most often been studied with corn oil as the vehicle of administration, the objectives of this study were to assess hepatotoxicity after exposure to single, low dosages of CHCl3 and BDCM given orally in an aqueous vehicle to estimate a lowest-observed adverse-effect level (LOAEL) and a no-observed-adverse-effect level (NOAEL) and to compare toxic potency. Ninety-day-old male Fischer 344 rats were gavaged with either 0.125, 0.1875, 0.25, 0.5, 0.75, 1.0, or 1.5 mmol CHCl3 or BDCM/kg body weight in 10% Alkamuls EL-620 (5 ml/kg body weight). At 24 h postgavage, serum was collected for analysis of clinical chemistry indicators of liver damage. Both CHCl3 and BDCM induced dose-dependent hepatotoxicity; serum alanine aminotransferase, aspartate aminotransferase, and sorbitol dehydrogenase were elevated significantly over control at 1.5, 1.0, and 0.5 mmol/kg. At these dose levels after 24 h, the two THMs appeared to be equipotent hepatotoxicants. Additional assessments at later time points demonstrated that BDCM causes more persistent liver damage than CHCl3 (Lilly et al., 1997). At 0.25, 0. 1875, and 0. 125 mmol of either THM/kg, significant increases over control were not detected for any measured endpoint. Therefore, these data indicate that the acute, oral NOAELs and LOAELs for liver toxicity are 0.25 and 0.5 mmol/kg, respectively, for both CHCl3 and BDCM. These determinations should provide a basis to establish new exposure limits for One-Day Health Advisories for these prevalent THMs. PMID- 9747605 TI - infoRAD at RSNA '98. Radiological Society of North America. PMID- 9747606 TI - I don't know, but I'll find out. PMID- 9747608 TI - CT in blunt chest trauma: indications and limitations. AB - Computed tomography (CT) is the imaging modality of choice in the assessment of patients with clinical or radiographic findings suggestive of aortic injury, bone fracture, or diaphragmatic tear following blunt chest trauma. Contrast material enhanced spiral CT allows detection of both subtle and more obvious aortic tears. CT has overall greater sensitivity than radiography in the detection of pulmonary lacerations and pneumothoraces. CT may be indicated in cases of suspected tracheobronchial injury. CT is of limited use in the assessment of rib fractures because such injuries are of limited clinical significance and can usually be identified at radiography; however, CT provides optimal visualization of thoracic spine fractures and superior assessment of suspected sternal fractures or sternoclavicular dislocation. Targeted spiral CT with sagittal and coronal reformatted images has increased sensitivity and specificity over that provided by conventional axial CT in the detection of diaphragmatic injury. Optimal CT assessment requires careful attention to technique, including the use of intravenously administered contrast material and multiplanar reconstructed images, as well as an awareness of potential pitfalls. Although in many cases diagnosis can be made with confidence on the basis of CT findings, further investigation is often needed to confirm the diagnosis. PMID- 9747607 TI - Imaging of mediastinal lymph nodes: CT, MR, and FDG PET. AB - The evaluation of mediastinal lymph nodes is an important aspect of staging in patients with non-small cell lung cancer. Anatomic imaging of lymph nodes with computed tomography (CT) and magnetic resonance (MR) imaging has been limited by the relatively low sensitivity and specificity of these techniques. Advances in physiologic imaging of mediastinal lymph nodes with 2-[fluorine-18] fluoro-2 deoxy-D-glucose (FDG) positron emission tomography (PET) have resulted in improved diagnostic accuracy in the determination of nodal status. Despite the limitations of CT, this technique still plays an important role by aiding in the selection of the most appropriate procedure for staging, by guiding biopsy, and by providing anatomic information for visual correlation with FDG PET images. At present, anatomic MR imaging of lymph nodes is primarily a problem-solving tool for cases with inconclusive CT results. Physiologic MR imaging with iron oxide is an exciting area of investigation, and the accuracy of this technique is being assessed in clinical trials. Anatomic and physiologic imaging techniques should be considered complementary rather than competitive imaging strategies. PMID- 9747609 TI - Radiographic and CT findings of blunt chest trauma: aortic injuries and looking beyond them. AB - Increasingly, helical CT is being used to screen trauma patients for aortic injury. Most aortic injuries visible at CT occur at or near the level of the ligamentum arteriosus; these injuries manifest as mediastinal hematoma, aortic contour deformity, intimal flaps, intraluminal debris, pseudoaneurysm, and pseudocoarctation. In the process of searching for aortic injury, however, the radiologist should not overlook other serious and more common thoracic injuries. Tracheobronchial tears appear at CT and radiography with persistent pneumothorax, subcutaneous emphysema, "fallen lung" sign, and malposition of endotracheal tube. The ruptured diaphragm, which tears more often on the left, appears asymmetric, irregular, or discontinuous, with herniation of bowel or viscera into the chest. In esophageal rupture, CT and radiography demonstrate left pneumothorax, pneumomediastinum, subcutaneous emphysema, and pleural effusion and atelectasis on the left. CT is better than trauma radiography for depicting fractures of the thoracic vertebral bodies and ribs, as well as for revealing pulmonary contusions and lacerations. CT is also useful for demonstrating unsuspected injuries caused by seat belts. Observation of these injuries should prompt a search for other serious internal organ injuries. PMID- 9747610 TI - Cartilaginous disorders of the chest. AB - Cartilaginous disorders of the thorax can arise in the parenchyma, airways, chest wall, and axial skeleton. At radiography, pulmonary hamartoma is characterized by "popcorn" calcification or fat density, either of which is diagnostic. Bronchiectasis is best demonstrated at high-resolution computed tomography (CT) and has a "tramline" or "signet ring" appearance. Tracheopathia osteochondroplastica appears at CT as multiple sessile submucosal nodules with or without calcification along the cartilaginous portion of the trachea. In relapsing polychondritis, the trachea and mainstem bronchi have diffuse or focal thickening with luminal narrowing at radiography. Costochondritis of the chest wall has become more prevalent with increased intravenous drug abuse and may be demonstrated at CT as soft-tissue swelling along with underlying cartilaginous fragmentation and bone destruction. Enchondromas are expansile and may display a calcified cartilaginous matrix at radiography. In osteochondroma, the thickness of the cartilaginous cap determines the likelihood of malignant degeneration. At radiography, chondroblastomas have a round contour, sharp margins, and cortical scalloping, whereas chondrosarcomas are large masses with indistinct margins, cortical breakthrough, and soft-tissue extension. By identifying either a process affecting a cartilage-containing structure or a cartilaginous matrix within a lesion, the chest radiologist may be able to narrow the list of differential diagnostic possibilities substantially. PMID- 9747611 TI - Radiation-induced changes in bone. AB - Radiation therapy has important applications in curative, adjuvant, and palliative therapy for a wide range of malignant conditions. Evidence of radiation therapy may be seen on radiologic images obtained subsequent to therapy. Bone growth disturbances may be observed in the immature axial or appendicular skeleton. Complications in the mature skeleton include osteoradionecrosis, pathologic fracture, and radiation-induced neoplasms. Radiologic features of mandibular osteoradionecrosis include ill-defined cortical destruction without sequestration. In osteoradionecrosis of the ribs, clavicle, scapula, and humerus, radiography may demonstrate osteopenia, disorganization and coarsening of trabecular architecture, and cortical irregularity; computed tomography more clearly depicts subtle fractures, alterations in bone architecture, and dystrophic soft-tissue calcification. In osteoradionecrosis of the spine, hematopoietic cellular elements of the spinal marrow are replaced with fat, which has high signal intensity on T1-weighted magnetic resonance images and intermediate signal intensity on T2-weighted images. Radiation-induced changes in the pelvis include osteopenia, increased bone density, and widening and irregularity of the sacroiliac joints. Radiation-induced osteochondromas are radiographically identical to those that arise spontaneously. Radiographic findings in radiation-induced sarcoma demonstrate an aggressive pattern of bone destruction. Awareness of the varied radiographic manifestations of radiation induced changes in bone and correlation with clinical features and the radiation field will usually allow distinction of these changes from those associated with other pathologic conditions. PMID- 9747612 TI - The false-negative mammogram. AB - In general, failure to detect or correctly characterize breast cancer can be attributed to one of four main factors: inherent limitations of screen-film mammography, inadequate radiographic technique, subtle or unusual lesion characteristics, and interpretation error. The restricted latitude and display contrast of screen-film mammography are among the significant factors that result in decreased visualization of breast tumors and microcalcifications in patients with dense fibroglandular tissue. Unlike the inherent limitations of screen-film mammography, a poor radiographic technique can be improved on and should be eliminated. Crucial components of a well-performed mammographic examination are correct positioning, adequate compression, and proper image exposure. Lesion characteristics that may lead to a false-negative mammogram include small size, a site where visualization is difficult, visualization on only one view, a benign or probably benign appearance, lack of a desmoplastic reaction, and slow or no apparent growth. Causes of interpretation error include suboptimal viewing conditions, outside distractions, lack of a systematic approach, oversight of a subtle lesion because of an obvious finding, lack of knowledge of clinical findings, imprecise correlation with results of other studies, and nonbelief. Recognition of these various factors should help decrease the rate of false negative mammograms. PMID- 9747613 TI - Translumbar placement of inferior vena caval catheters: a solution for challenging hemodialysis access. AB - Access to the central venous circulation for hemodialysis has traditionally been achieved via the subclavian or jugular venous routes. With ongoing improvements in medical management, many hemodialysis recipients develop exhaustion of these routes and require alternative means of central venous access. Inferior vena caval (IVC) catheters have been placed with a percutaneous translumbar approach to allow central venous access for chemotherapy, harvesting of stem cells, and total parenteral nutrition. Translumbar placement of IVC catheters has become accepted by some as a useful and reliable alternative in patients who require long-term hemodialysis but have exhausted traditional access sites. IVC catheters have been placed in patients with IVC filters, and IVC filters have been placed in patients with IVC catheters. Complications include those associated with central venous catheters, for example, sepsis, fibrin sheaths, and thrombosis. A complication specific to placement of IVC hemodialysis catheters is migration of the catheter into the subcutaneous soft tissues, retroperitoneum, or iliac veins. Translumbar placement of IVC catheters is performed only in patients considered to have few or no other medical options and is not intended as a primary means of central venous access. PMID- 9747614 TI - Pancreatic disease in children and young adults: evaluation with CT. AB - In children with pancreatic disease, computed tomography (CT) has a primary role in the evaluation of pancreatitis, trauma, and malignancy. At CT, pancreatic abnormalities may manifest as pancreatic enlargement (tumor, acute pancreatitis), pancreatic atrophy (cystic fibrosis, chronic pancreatitis), cystic lesions (pseudocysts, congenital simple cysts, autosomal dominant polycystic kidney disease, von Hippel-Lindau disease, cystic fibrosis, cystic neoplasms), or fatty replacement (cystic fibrosis, Shwachman-Diamond syndrome, history of steroid therapy, Cushing syndrome, Johanson-Blizzard syndrome, obesity). CT is the best modality for evaluation of pancreatitis, allowing detection of pancreatic abnormalities as well as abnormal extrapancreatic fluid collections. In children who have undergone blunt abdominal trauma, CT has been shown to be the best initial imaging study, being more sensitive than ultrasound for detection of pancreatic injury. In neoplastic conditions, CT demonstrates the extent of disease, enables characterization of the tissue components of the tumor, and allows accurate posttreatment follow-up. Although the various diseases of the pancreas may have overlapping appearances at CT, the correct diagnosis can often be made on the basis of the CT findings in combination with the clinical history, laboratory data, and the patient's age. PMID- 9747615 TI - Non-squamous cell neoplasms of the larynx: radiologic-pathologic correlation. AB - A variety of benign and malignant non-squamous cell neoplasms may affect the larynx. Most of these uncommon laryngeal neoplasms are located beneath an intact mucosa, making diagnosis difficult with endoscopy alone, and sampling errors may occur if only traditional superficial biopsies are performed. In some laryngeal neoplasms, radiologic evaluation allows the correct diagnosis. Hemangiomas have very high signal intensity at T2-weighted magnetic resonance (MR) imaging and strong enhancement at both computed tomography (CT) and MR imaging after administration of contrast material. Phleboliths, which are pathognomonic for hemangiomas, are easily identified at CT. Chondrogenic tumors typically manifest with coarse or stippled calcifications at CT. Because of their high water content, chondrogenic tumors have very high signal intensity on T2-weighted MR images, whereas only moderate enhancement is observed after administration of contrast material. Lipomas typically manifest at both CT and MR imaging as homogeneous nonenhancing lesions. They are isoattenuating to subcutaneous fat at CT and isointense relative to subcutaneous fat with all MR pulse sequences. Metastases from renal adenocarcinoma typically demonstrate strong contrast enhancement and flow voids at MR imaging, and metastases from melanotic melanoma usually have high signal intensity on T1-weighted MR images and low signal intensity on T2-weighted images owing to the paramagnetic properties of melanin. Although radiologic findings are nonspecific in most other non-squamous cell neoplasms of the larynx (eg, Kaposi sarcoma, hematopoietic tumors, tumors of the minor salivary glands, metastases from amelanotic melanoma), cross-sectional imaging can play an important role in the diagnostic work-up of these unusual tumors by delineating the extent of submucosal tumor spread and directing the endoscopist to the appropriate site for the deep, transmucosal biopsies needed to establish the diagnosis. In addition, CT and MR imaging are crucial for posttherapeutic monitoring and early detection of local recurrence. PMID- 9747616 TI - Enchondroma versus chondrosarcoma in the appendicular skeleton: differentiating features. AB - Distinction of enchondroma versus intramedullary chondrosarcoma affecting the appendicular skeleton (proximal to the metacarpals and metatarsals) is a frequent diagnostic dilemma. The authors studied a large series of patients with these lesions (92 with enchondromas, 95 with chondrosarcomas) using statistical assessment of both clinical parameters and numerous radiologic manifestations on images from multiple modalities to identify differentiating features. Multiple clinical and imaging parameters demonstrated statistically significant differences between enchondroma and chondrosarcoma, particularly pain related to the lesion, deep endosteal scalloping (greater than two-thirds of cortical thickness), cortical destruction and soft-tissue mass (at computed tomography or magnetic resonance imaging), periosteal reaction (at radiography), and marked uptake of radionuclide (greater than the anterior iliac crest) at bone scintigraphy. All of these features strongly suggested the diagnosis of chondrosarcoma. These criteria allow distinction of appendicular enchondroma and chondrosarcoma in at least 90% of cases. PMID- 9747617 TI - Radioactive decay. AB - When a parent radionuclide decays to its daughter radionuclide by means of alpha, beta, or isomeric transition, the decay follows an exponential form, which is characterized by the decay constant lambda. The decay constant represents the probability per unit time that a single radioatom will decay. The decay equation can be used to provide a useful expression for radionuclide decay, the half-life, the time when 50% of the radioatoms present will have decayed. Radiotracer half life has direct implications in nuclear imaging, radiation therapy, and radiation safety because radionuclide half-life affects the ability to evaluate tracer kinetics and create appropriate nuclear images and also affects organ, tumor, and whole-body radiation dose. The number of radioatoms present in a sample is equal to the activity, defined as the number of transitions per unit time, divided by the decay constant; the mass of radioatoms present in a sample can be calculated to determine the specific activity (activity per unit mass). The dynamic relationship between the number of parent and daughter atoms present over time may lead to radioactive equilibrium, which takes two forms--secular and transient -and has direct relevance to generator-produced radionuclides. PMID- 9747618 TI - Phase-contrast radiography. AB - For the past 100 years, the paradigm for radiography has been premised on absorption as the sole means of contrast formation and on ray optics as the basis for image interpretation. A new conceptual approach to radiography has been developed that includes phase (ie, refractive) contrast and requires wave optics for proper treatment. This new approach greatly increases the amount of information that can be obtained with radiographic techniques and is particularly well suited to the imaging of soft tissue and of very small features in biologic samples. A key feature of the present technique of phase-contrast radiography is the use of a microfocus x-ray source about an order of magnitude (< or = 20 microm) smaller than that used in conventional radiography. Phase-contrast radiography offers a number of improvements over conventional radiography in a clinical setting, especially in soft-tissue imaging. These improvements include increased contrast resulting in improved visualization of anatomic detail, reduced absorbed dose to the patient, inherent image magnification and high spatial resolution, use of harder x rays, and relative ease of implementation. More technologically advanced detectors are currently being developed and commercialized, which will help fully realize the considerable potential of phase contrast imaging. PMID- 9747619 TI - MR diffusion imaging in stroke: review and controversies. AB - Magnetic resonance (MR) diffusion imaging allows detection of cerebral ischemia within minutes of onset, and the temporal evolution of diffusion characteristics enables differentiation of acute from chronic stroke. T2-weighted MR imaging demonstrates infarcted tissue but fails to demonstrate acutely ischemic regions. Furthermore, the similar signal intensity characteristics of acute and chronic stroke on T2-weighted images limit the ability to determine the acuteness of an infarct. Diffusion imaging thus has tremendous potential for helping direct the treatment of acute ischemic stroke. Controversy exists over the pathophysiology of underlying changes in diffusion and the reversibility of changes after reperfusion in humans. There is also a lack of reproducibility in the time course of diffusion changes between research centers. Use of optimal diffusion imaging strategies results in increased conspicuity of ischemic regions and increased reproducibility of diffusion constants between research centers. An understanding of the principles of diffusion imaging and current controversies in the field is necessary for optimal application of this technique in the evaluation and treatment of cerebral ischemia. PMID- 9747620 TI - Enhancing Web applications in radiology with Java: estimating MR imaging relaxation times. AB - Java is a relatively new programming language that has been used to develop a World Wide Web-based tool for estimating magnetic resonance (MR) imaging relaxation times, thereby demonstrating how Java may be used for Web-based radiology applications beyond improving the user interface of teaching files. A standard processing algorithm coded with Java is downloaded along with the hypertext markup language (HTML) document. The user (client) selects the desired pulse sequence and inputs data obtained from a region of interest on the MR images. The algorithm is used to modify selected MR imaging parameters in an equation that models the phenomenon being evaluated. MR imaging relaxation times are estimated, and confidence intervals and a P value expressing the accuracy of the final results are calculated. Design features such as simplicity, object oriented programming, and security restrictions allow Java to expand the capabilities of HTML by offering a more versatile user interface that includes dynamic annotations and graphics. Java also allows the client to perform more sophisticated information processing and computation than is usually associated with Web applications. Java is likely to become a standard programming option, and the development of stand-alone Java applications may become more common as Java is integrated into future versions of computer operating systems. PMID- 9747621 TI - Giant cell tumor of the skull. PMID- 9747622 TI - General case of the day. End-stage sarcoidosis. PMID- 9747623 TI - US case of the day. Diffuse large B-cell lymphoma (posttransplant lymphoproliferative disorder [PTLD]). PMID- 9747624 TI - Breast imaging case of the day. Intracystic papillary carcinoma of the breast. PMID- 9747625 TI - Neuroradiology case of the day. Burkitt lymphoma. PMID- 9747626 TI - Pediatric case of the day. Prune-belly syndrome (Eagle-Barrett syndrome, triad syndrome). PMID- 9747627 TI - Neoplasms associated with Paget disease. PMID- 9747628 TI - Paper radiologic images. PMID- 9747629 TI - Advances in immunotherapy of hematologic malignancies. AB - The chimeric anti-CD20 antibody rituxamab, as well as radiolabeled anti-CD20 monoclonal antibodies, have demonstrated significant activity against B-cell non Hodgkin's lymphoma. Idiotype vaccination in remission may prevent relapse in follicular non-Hodgkin's lymphoma. The campath-1H antibody has activity in chronic lymphocytic leukemia, and additional unconjugated, radiolabeled, and drug conjugated monoclonal antibodies (anti-CD45 and anti-CD33) have shown activity in acute myeloid leukemia. Adoptive cellular therapy is active against posttransplantation relapse and lymphoproliferative disorders in most patients, and complications of graft-versus-host disease may be controlled by suicide gene transfection of the donor lymphocytes. PMID- 9747630 TI - Chronic lymphocytic leukemia. AB - Recent studies have improved our understanding of the cytogenesis, biology, and therapy of chronic lymphocytic leukemia (CLL). This review highlights this recent progress reported over the past year. We have improved our understanding of the cytogenetic abnormalities in CLL and soon may see identification of new tumor suppressor genes that may be deleted in the leukemia cells of a large number of patients with this disease. We have achieved a better understanding of the surface antigens that help govern the pattern of tissue-infiltration of leukemia cells in vivo. Studies on the immune pathophysiology of CLL are providing clues to potential mechanisms leading to the immunodeficiency associated with this disease. Combination chemotherapy with purine analogues is showing promise for improved efficacy in CLL. Finally, new therapies incorporating bone marrow transplantation, and possibly gene therapy, increasingly are being considered for the therapy of patients with this disease. PMID- 9747631 TI - Recent advances in the treatment of multiple myeloma. AB - Multiple myeloma represents the second most common hematologic malignancy, with nearly 15,000 new cases each year in the United States. Although further understanding of the pathogenesis of this B-cell malignancy has been made, the disease remains incurable with a median survival of approximately 3 years. The identification of new genetic events in the malignant cells themselves may lead to new potential therapies. Moreover, recent identification of the new human herpesvirus 8 in the supporting cells of the bone marrow of these patients will likely change approaches to this disease in the laboratory and the clinic. Further development of new high-dose therapy approaches has led to a reduction in treatment-related mortality with an improvement in overall survival. Treatment with the bisphosphonate pamidronate reduces skeletal complications and may also improve overall survival of these patients. PMID- 9747632 TI - Advances in the management of Hodgkin's and non-Hodgkin's lymphoma. AB - Recent developments in the management of lymphoma continue to refine our approach to the disease. There is increasing acceptance of the International Lymphoma Study Group classification for non-Hodgkin's lymphoma. Identification of a new viral agent associated with certain subtypes of lymphoma and new immunologic therapies change our perspective on the disease. New data on treatment results further elucidate the role of radiotherapy and high-dose therapy in Hodgkin's disease and the non-Hodgkin's lymphomas. PMID- 9747633 TI - Molecular basis of leukemogenesis. AB - Somatically acquired translocations play a major role in the pathogenesis of human leukemias. These rearrangements frequently alter the structure or expression of genes encoding key regulatory proteins involved in normal hematopoiesis. Analysis of these proteins in the human leukemias and in genetically manipulated mouse models has greatly increased our understanding of the mechanisms of leukemogenesis. This information also has led to improvements in our ability to deliver the optimal treatment intensity to individual patients and to the development of novel leukemia-specific therapies. PMID- 9747635 TI - Multidrug resistance in leukemia. AB - Resistance of tumors to chemotherapeutic agents is an important factor that limits the successful treatment of a wide range of malignancies. The multidrug resistant (MDR) phenotype is well recognized in clinical samples, and it has been extensively studied, particularly in acute myeloid leukemia. One of the principle mechanisms underlying this MDR phenotype is the active cellular extrusion of chemotherapeutic agents by the multidrug resistance protein MDR1 (p glycoprotein). More recently, other drug resistance proteins, notably the multidrug resistance-associated protein, MRP, and the lung resistance protein, LRP, have also been implicated in multidrug resistance. This review summarizes recent contributions in this field. In particular, it focuses on recent efforts to better measure MDR in clinical samples, a critical step in correctly determining the MDR phenotype of a patient's tumor. PMID- 9747634 TI - Supportive care in hematologic malignancies. AB - Supportive care in hematologic malignancies includes a wide range of topics. We have selected the following issues for a review of recently published developments: new insights into the benefits and risks of established hematopoietic growth factors (HGF), such as granulocyte- or granulocyte macrophage colony-stimulating factor (G-CSF, GM-CSF); the emerging role of newly introduced HGFs such as keratinocyte-growth factor (KGF) and thrombopoietin; the prophylactic and therapeutic use of amifostine, a cytoprotective agent; the role of hematopoietic growth factors and the demethylating agent decitabine in myelodysplastic syndromes (MDS); infectious complications of anticancer therapy; and, recent improvements in and complications of transfusional therapy, including the renewed interest in granulocyte transfusions. PMID- 9747636 TI - Recent advances in the biology and treatment of childhood acute lymphoblastic leukemia. AB - As cure rates in childhood acute lymphoblastic leukemia edge toward 80%, the focus of research is shifting to better means of identifying and treating resistant cases. This new emphasis has stimulated progress in several areas. Recent findings suggest that poor early responses to therapy and detection of minimal residual disease at the postremission induction period by immunologic methods are reliable indicators of an adverse prognosis warranting modification of treatment. In this regard, timely administration of intensified chemotherapy, including a second reinduction/intensification phase, may nullify the adverse prognosis conferred by a delayed response to induction therapy. Comparative analysis of survival outcomes in T-cell patients who received chemotherapy or cranial irradiation (12 Gy) to prevent overt leukemia in the central nervous system suggests that the latter modality should be retained for cases with leukocyte counts > 100 x 10(9)/L. Recent innovations in histocompatibility matching, prevention of graft-versus-host disease, and antiviral prophylaxis have enhanced the applicability of hematopoietic stem cell transplantation, making this procedure available to candidates lacking matched sibling donors. Finally, demonstration that acute lymphoblastic leukemia has an angiogenic phase in bone marrow raises the possibility of effective treatment with antiangiogenic agents, such as endostatin. Remaining challenges in the treatment of childhood leukemia include 1) the development of specific and more effective therapy for high-risk cases and 2) the reduction of long-term complications associated with intensive chemotherapy and cranial irradiation. PMID- 9747637 TI - Chronic myeloid leukemia. AB - Progress has been made in understanding BCR-ABL-positive leukemias. A new transcript (p230BCR-ABL) has been characterized that is associated with Ph positive chronic neutrophilic leukemia. The ATM protein appears to be a regulator of ABL activity in response to irradiation damage. Pathways linking BCR-ABL to the BCL-2 family of proteins may be active in Philadelphia-positive cells and inhibit apoptosis. The 62-kD protein constitutively phosphorylated in chronic myeloid leukemia progenitors has been cloned. Ph-negative long-term culture initiating cells are detectable in many chronic myeloid leukemia patients. The combination of interferon alfa and cytarabine appears to be superior to interferon alfa alone. Autografting with in vivo-purged stem cells may induce prolonged remissions. Specific inhibitors of the BCR-ABL tyrosine kinase are becoming available. PMID- 9747638 TI - Current world literature. Hematologic malignancies. PMID- 9747639 TI - The belief in conventional medicine and adherence to treatment in non-insulin dependent diabetes mellitus patients. AB - We investigated the role of belief in conventional medicine, the type of medical care, and familiar and socioeconomic factors on the adherence to treatment in non insulin-dependent diabetes mellitus (NIDDM) patients. In a cross-sectional design, we selected 156 patients from two institutions, who agreed to fill out a questionnaire, which included general data, socioeconomic level, somatometric data, type of medical care, complications, if they had friends and relatives with diabetes, the family function, and a score on the belief in conventional medicine. Factors associated with adherence to diet and medication were analyzed. Patients had a mean age of 55.6 years and 8.9 years since diagnosis. A total of 51.3% of them were not covered by social security, and 62.8% received attention by a general physician. Patients under the care of a specialist had better adherence to diet and medication, and better belief in conventional medicine. The principal factor associated with adherence to medication and diet was the belief in conventional medicine (p < 0.001 in both). Adherence to diet was also associated with the socioeconomic level (p=0.001) and years since diagnosis (p=0.004). Adherence to medication was also associated with schooling (p=0.001). We concluded that belief in conventional medicine is strongly associated with adherence to treatment and other factors such as schooling, socioeconomic level, and medical care under a specialist; adherence to diet was better in patients with more years since diagnosis. PMID- 9747640 TI - Survival following alpha particle pituitary irradiation for diabetic retinopathy. AB - The purpose of this study was to evaluate the mortality experience of persons with longstanding diabetes who had received pituitary irradiation for diabetic retinopathy compared to a matched group of persons with diabetes who had not had pituitary ablation. The irradiated cohort consisted of 167 patients treated at the Donner Pavilion (Lawrence Berkeley Laboratory, University of California, Berkeley), and the comparison cohort was the population evaluated in the Wisconsin Epidemiologic Study of Diabetic Retinopathy (WESDR). Survival analyses were performed comparing the two cohorts using three different sets of matching criteria, each more restrictive than the previous analyses. The three different strategies were (1) matched only on severity of diabetic retinopathy; (2) matched on severity of retinopathy and age; and (3) matched on severity of retinopathy, age, gender, and hypertension status. Tests of comparison were the log-rank test, the Wilcoxon test, and the likelihood ratio test. For the model matching only on severity of retinopathy, mean survival was 8.3 years for the WESDR group and 9.4 years for the ablated group (p > 0.05 for all three statistical tests). For the model matched on retinopathy and age, mean survival was 8.9 years for the WESDR group and 9.2 years for the ablated group (p=0.05 log-rank test, 0.32 Wilcoxon test, and 0.06 likelihood ratio test). For the model matching on retinopathy, age, gender, and hypertension status, mean survival was 8.9 years for the WESDR group and 11.6 years for the ablated group (p=0.72 log-rank test, 0.08 Wilcoxon test, and 0.82 likelihood ratio test). These data are compatible with the notion that pituitary ablation, and therefore induced pituitary growth hormone deficiency, may not decrease survival in those with severe diabetic retinopathy. PMID- 9747641 TI - Reduced expression of a novel peptide, prostacyclin-stimulating factor, in the kidneys of streptozotocin-induced diabetic rats. AB - Prostacyclin (PGI2) produced by vascular endothelial cells (ECs) is a potent vasoactive prostanoid involved in maintenance of vessel wall homeostasis. Reduced PGI2 synthesis by vascular ECs could be a mechanism of pathogenesis in the development of vascular lesions such as diabetic angiopathy. Recently, we purified and cloned a novel bioactive peptide, PGI2-stimulating factor (PSF), which stimulates PGI2 production by vascular ECs. PSF may act on vascular ECs in a paracrine and/or autocrine fashion to regulate PGI2 synthesis. Decreased PSF production in the vessel wall may result in an imbalance of prostanoid synthesis, leading to the development of vascular lesions such as diabetic angiopathy. Our immunohistochemical study demonstrated that PSF is located in vascular resident cells such as vascular smooth muscle cells (SMCs) and ECs, as well as in bronchial SMCs. Moreover, PSF mRNA was found to be expressed in various tissues in Wistar rats, particularly in the kidneys and lungs. The present study demonstrated that streptozotocin (STZ)-induced diabetic rats showed less PSF mRNA expression in the kidneys (PSF mRNA/28S rRNA ratio; STZ versus control; 1.7+/-0.2 versus 2.5+/-0.2, p < 0.05) and reduced immunohistochemical staining for PSF in arteries in the kidney. However, in the lungs, there were no changes in tissue PSF mRNA expression (STZ versus control; 10.9+/-0.9 versus 11.5+/-1.0, NS) or in the extent of PSF staining in bronchial SMCs of STZ-induced diabetic rats. These findings suggest that decreased expression of PSF in renal vessels of STZ-induced diabetic rats may cause an imbalance of prostanoid synthesis, leading to the development and progression of vascular damage in the kidney. PMID- 9747642 TI - Ascorbic acid clearance in diabetic nephropathy. AB - The incidence of cardiovascular disease is increased in diabetic nephropathy. Increased oxidative stress in diabetes is believed to play an important role in the pathogenesis of atherosclerosis in diabetes. Since antioxidant vitamins, such as ascorbic acid, often are reduced in diabetes, we hypothesized that the renal clearance of ascorbic acid is increased in patients with diabetic nephropathy. Thirty-seven subjects with diabetic nephropathy were studies: 18 had microalbuminuria (30-300 mg/day albuminuria); the remainder had clinical nephropathy (> 300 mg/day albuminuria). Indices of glycemic control (glucose, hemoglobin A1C) and renal function (albuminuria and creatinine clearance) were measured in addition to serum and urinary ascorbic acid levels. Results showed that subjects with clinical nephropathy had lower mean plasma ascorbic acid (p=0.0009) and higher renal clearance of ascorbic acid (p=0.005) than those with microalbuminuria. Bivariate analysis revealed an inverse correlation between creatinine clearance and AA clearance (r=-0.42, p=0.009). There was a significant linear association between the quantity of albuminuria and ascorbic acid clearance (r=0.49, p=0.002). Thus, patients with diabetic nephropathy have reduced ascorbic acid levels due to increased ascorbic acid clearance. The decrease in antioxidant defense that arises from the low levels of vitamin C may contribute to the increased cardiovascular morbidity and mortality observed in this population. PMID- 9747643 TI - Decreased urinary kallikrein with hyperglycemia in patients with short-term insulin-dependent diabetes mellitus. AB - The aim of the study was to evaluate the role of urinary kallikrein in the regulation of renal hemodynamics and sodium handling in insulin-dependent diabetes mellitus (IDDM), and to test the effect of acutely induced hyperglycemia. Urinary kallikrein excretion was evaluated (1) under basal conditions and after stimulation with i.v. furosemide (0.5 mg x kg(-1)), (2) during glycemic clamp-induced eu- and hyperglycemia (5 and 12 mmol/L) and, (3) during time-controlled euglycemia in 21 short-term IDDM patients without microalbuminuria and in 18 weight-, age- and gender-matched healthy controls. Sodium excretion and renal hemodynamics using the clearances of inulin and para amino-hippuric acid were measured during examinations in both groups. The baseline urinary kallikrein excretion during clamp-induced euglycemia was comparable in diabetic and control subjects (10.89+/-5.98 versus 10.38+/-3.73 mUE x min(-1)), whereas it was decreased in the baseline for furosemide (5.77+/-3.22 versus 10.9+/-3.7 mUE x min(-1); p < 0.01) and even after furosemide administration (12.0+/-1.6 versus 21.3+/-2.0 mUE x min(-1); p < 0.01) while the patients were hyperglycemic. During intravenous dextrose-induced hyperglycemia, the urinary kallikrein excretion significantly declined in diabetic patients (10.89+/-5.98 versus 5.45+/-0.88 mUE x min(-1); p < 0.01), whereas it did not change in controls (10.38+/-3.73 versus 12.55+/-5.47 mUE x min(-1)). A decrease in the fractional excretion of sodium and an attenuated rise in natriuresis after furosemide administration have been found in diabetic compared to control subjects. There were no significant relationships between kallikrein excretion and (1) renal hemodynamics, which was comparable in both groups, or (2) plasma renin activity, plasma and urine aldosterone and cortisol. We conclude that short term IDDM without renal hemodynamic alterations is associated with decreased basal and furosemide-stimulated kallikrein excretion, which is directly related to the blood glucose level. The decreased activity of the renal kallikrein-kinin system might be involved in the increased tendency to sodium retention in diabetic patients. PMID- 9747644 TI - Lipid- and glucose-lowering efficacy of Plantago Psyllium in type II diabetes. AB - The beneficial effect of dietary fiber in the management of type II diabetes is still controversial and has not been totally demonstrated. The purpose of this study was to determine the plasma-lowering effects of 5 g t.i.d. of Plantago Psyllium, as an adjunct to dietary therapy, on lipid and glucose levels, in patients with type II diabetes. Patients were randomly selected from an outpatient clinic of primary care to participate in a double-blind placebo controlled study in which Plantago Psyllium or placebo was given in combination with a low fat diet. One hundred twenty-five subjects were included in the study that consisted in a 6-week period of diet counseling followed by a 6-week treatment period. Fasting plasma glucose, total plasma cholesterol, LDL cholesterol, HDL cholesterol and triglyceride levels were measured every 2 weeks. The test products (Psyllium or placebo) were supplied to subjects in identically labeled foil packets containing a 5-g dose of product, to consume three doses per day (of 5 g each one), before regular meals. There was an excellent tolerance to Psyllium, without significant adverse effects. No significant changes were observed in the patient's weight for both groups (not significant). Fasting plasma glucose, total cholesterol, LDL cholesterol, and triglycerides levels, showed a significant reduction (p < 0.05), whereas HDL cholesterol increased significantly (p < 0.01) following Psyllium treatment. Our results show that 5 g t.i.d. of Psyllium is useful, as an adjunct to dietary therapy, in patients with type II diabetes, to reduce plasma lipid and glucose levels, resolving the compliance conflict associated with the ingest of a great amount of fiber in customary diet. PMID- 9747645 TI - Patient and family reflections on the use of subcutaneous insulin to prevent diabetes: a retrospective evaluation from a pilot prevention trial. AB - A retrospective, semi-structured telephone interview was used to collect data from 28 of 31 families who participated in a pilot study testing subcutaneous insulin as a means to prevent or delay insulin-dependent diabetes mellitus (IDDM) onset. Interviews were conducted an average of 3 years after the initiation of the subcutaneous insulin protocol. Both the high-risk person (if > or = 8 years of age) and a family member (spouse or parent) were interviewed. Most participants reported that they were distressed to learn that they or a family member were at risk for IDDM, and families readily agreed to initiate subcutaneous insulin therapy. More children than adults reported insulin injections and blood glucose tests were "hard" or "very hard," but noncompliance was more common in adults. Of the high-risk participants interviewed, 58% of children and 100% of adults reported experiencing hypoglycemia, although episodes requiring someone else's assistance were rare, occurring in 38% of the children and 27% of the adults. Participants remained enthusiastic about trial participation; most favored screening programs to identify those at risk for IDDM, believed screening should be conducted regardless of age, believed subcutaneous insulin prevented or delayed IDDM onset, and would recommend subcutaneous insulin therapy to another high-risk individual. In addition, more than 40% of children, parents, and spouses reported that they would have benefited from access to a mental health professional at some point during the trial. PMID- 9747646 TI - Acute Charcot arthropathy in patients with diabetes mellitus: healing times by foot location. AB - Foot deformity and lower extremity dysfunction are debilitating complications of diabetes mellitus which often lead to significant permanent disability. Acute diabetic neuroarthropathy (Charcot arthropathy) directly leads to foot deformity, subsequent lower-extremity complications and may lead to lower-extremity amputation, if not identified and managed appropriately. The purpose of this study is to report the healing times of acute-onset neuropathic arthropathies (fractures, joint subluxations or dislocations) in individuals with diabetes mellitus by foot location using the ambulatory method of total-contact casting (TCC). In addition, the identification of critical subject characteristic which influence healing outcomes were determined. The results indicate all (100%) of the acute (Charcot) fractures, subluxations, or dislocations healed in an average of 86+/-45 days. Acute Charcot arthropathies of the ankle, hindfoot, or midfoot take longer to heal by TCC than arthropathies localized to the forefoot. Adherence to partial weight bearing with assistive devices during casting and early institution of cast immobilization are critical factors associated with shorter healing times using TCC. Physicians, rehabilitation specialists and third party payers should be aware of the length of time required to heal acute Charcot foot arthropathies at all locations of the foot using TCC. PMID- 9747647 TI - Effects of mild chronic heat exposure on the concentrations of thiobarbituric acid reactive substances, glutathione, and selenium, and glutathione peroxidase activity in the mouse liver. AB - To determine whether mild and chronic heat stress leads to oxidative stress and to differentiate such effects of different exposure periods, we kept male ICR mice at an ambient temperature of either 35 degrees C or 25 degrees C for 6 hours, 3 days, or 7 days and measured the concentrations of thiobarbituric acid reactive substances (TBARS), glutathione (GSH), selenium (Se), and glutathione peroxidase (GSH-Px) activities in the liver. Since the food consumption of the heat-exposed group was only half that of the control, we prepared pair-fed groups, which were kept at 25 degrees C and whose food consumption were limited to those of the heat-exposed group for the 3-day and the 7-day exposure. TBARS concentrations of the liver was significantly higher in the heat group than the control after the 3-day exposure, while there was no significant difference among the groups after the 7-day exposure. There was no significant difference in GSH concentrations between the heat-exposed group and the control after the 7-day exposure, when the GSH concentration of the pair-fed group was significantly lower than that of the control. Hepatic cytosolic Se GSH-Px activity in the heat group was significantly less than that in the control group after the 6-hour exposure and it tended to be lower in the heat group than that of the control group after the 7-day exposure, while there was no difference in the total GSH-Px activity among the three groups. Our results showed that mild and chronic heat exposure may cause oxidative damage to organisms and that GSH-related anti oxidative systems would play an important role to defensive reaction. PMID- 9747648 TI - Biodistribution study of murine monoclonal anti-GD3 antibody in nude mice bearing human melanoma xenografts for development of immunoscintigraphy. AB - Reactivity of the monoclonal antibody with the tumor markers is known to be different between cultured cells in vitro and transplanted tumors in vivo. The monoclonal antibody should be investigated regarding its specific accumulation in tumor-bearing mice for immunodetection or immunotherapy. We studied the biodistribution of radiolabeled monoclonal anti-GD3 antibody (IgM) in normal mice and nude mice bearing human melanoma xenografts. Tissue-to-blood distribution ratios of the antibody in the liver, spleen and kidney increased with time in both normal and melanoma-transplanted mice, but no significant changes were observed in other normal tissues up to 5 days after injection. Specific accumulation of the monoclonal anti-GD3 antibody in the grafted human melanoma (HMV-II) was observed 4 and 5 days after injection. On the other hand, no specific accumulation of standard murine IgM in the tissue of HMV-II was observed in mice bearing the HMV-II xenograft 5 days after injection. Because the tissue to-blood ratio of the distribution in the tissue of HMV-II became larger than that of other tissues 4 and 5 days after administration, 4 days after the administration of the monoclonal anti-GD3 antibody were required for immunoscintigraphy. Accumulation of the monoclonal anti-GD3 antibody in other human melanomas (HMV-I, HMY-1 and SK-MEL188) inoculated into mice was also observed 4 days after the antibody administration. The monoclonal anti-GD3 antibody used in this study would be useful in immunodetection or immunotherapy. PMID- 9747649 TI - The effect of short-term octreotide administration on the histologic structure of stomach, duodenum, jejunum, colon, liver and gallbladder in an experimental pancreatitis model. AB - It is known that long-term administration of octreotide leads to changes in the histology of intraabdominal organs and plasma biochemical values. The purpose of the present study was to evaluate the histological effect of short-term octreotide administration on digestive organs in the experimentally induced pancreatitis by ligating pancreatic duct. The sham operation was performed on 20 rabbits in Groups 1 and 2. Acute pancreatitis was induced by pancreatic duct ligation in 20 rabbits in Groups 3 and 4. Octreotide was administered subcutaneously to the rabbits in Groups 2 and 4 at a dosage of 10 microg/kg/day for 7 days. The animals were sacrificed at the end of day 7, blood and tissue samples were collected. There was no histological changes in the stomach, duodenum, gallbladder, or small and large intestines of those group which received octreotide, while hepatic bile duct proliferation, bile duct epithelium proliferation, periportal inflammation and venous stasis were observed in liver histology. In conclusion, one-week octreotide administration in this experimental acute pancreatitis model was not associated with pathologic changes in digestive organs except liver. PMID- 9747650 TI - Age-related effects of rolipram on [3H]quinuclidinyl benzilate and [3H]phorbol 12,13-dibutyrate binding in the rat brain. AB - Cholinergic neurotransmission and protein kinase C (PKC) in the brain play important roles in the processes of cognitive function. In this study, we examined the effect of chronic treatment with rolipram, a 3',5'-cyclic adenosine monophosphate (cyclic AMP)-selective phosphodiesterase inhibitor, on age-related changes in [3H]quinuclidinyl benzilate (QNB) and [3H]phorbol 12,13-dibutyrate (PDBu) binding, which labeled brain muscarinic cholinergic receptors and PKC, respectively. Rolipram was administered per os to young (15 weeks old) and old (80 weeks old) Wistar rats at dosage of 0.01 mg/kg and 0.1 mg/kg once a day over 4 weeks. Then, quantitative in vitro autoradiography was performed. Control old rats showed elevations in [3H]PDBu binding in the hippocampus and the cerebellum compared to young rats, but [3H]QNB binding was largely unchanged. Chronic treatment of the old rats with the higher dose of rolipram led to reductions in [3H]QNB and [3H]PDBu binding in many brain regions. However, the same treatment of the young rats induced no or minimal effect. Thus, the response of the brain to rolipram was different between young and old rats. These results suggest that the cyclic AMP-selective phosphodiesterase system in the brain is modified during aging, modulating subsequently cholinergic neurotransmission and PKC activity exclusively in old rat brains. PMID- 9747651 TI - Experimental ablation of emphysematous rat lung with Nd: YAG laser: lung changes studied by histopathology and SEM. AB - Laser ablation has been employed as a therapeutic measure for chronic pulmonary emphysema. As yet, however, its effect is not understood on firm pathological basis. We aimed to study, both histopathologically and using Scanning Electric Microscopy (SEM), the changes produced by irradiation with contact Neodymium yttrium aluminum garnet laser (Nd: YAG laser) in rat lungs with experimentally induced emphysema. Emphysema was produced in 34 rats by instilling elastase via airways. Eight weeks after the instillation, the emphysematous left lung was irradiated under thoracotomy with contact Nd: YAG laser at a power of 5 watts. The animals were sacrificed in acute as well as chronic phase for histopathological observation of lung and scanning electron microscopy. Laser caused necrotic and inflammatory changes in the subpleural zone of lung. Immediately after irradiation, the alveolar septa were destroyed as visualized by SEM, only leaving the elastic skeleton. In a chronic phase, the necrotic zone was collapsed and replaced with a thick fibrous scar which seemed to serve more or less to keep the organ from being excessively inflated. In this model, irradiation induces subpleural dense scarring, which, by "encasing" an emphysematous lung, is expected to more or less normalize the excessive compliance. PMID- 9747652 TI - The acoustic reflex for filtered broadband stimuli: a lesser contribution of the lower frequency neurons. AB - The effects of the cut-off frequency of the filtered broadband stimuli on the human acoustic reflex (AR) were examined to observe the relation between the area of excitation in the cochlea and the AR response. The results obtained have indicated that all the input from the cochlear region does not equally contribute to trigger the AR equally; i.e., there is a lesser contribution from the frequency region below 700 Hz. PMID- 9747653 TI - Differential localization of mRNAs for mammalian trps, presumptive capacitative calcium entry channels, in the adult mouse brain. AB - Mammalian homologues for Drosophila trp are likely to be candidates for capacitative calcium entry channels. By in situ hybridization histochemistry, we have demonstrated that the mRNAs for four species of the mouse homologues (Mtrps 1, -3, -4, -6) were differentially expressed in the adult mouse brain. Mtrp-1 mRNA was expressed widely throughout the gray matters, while the expression for Mtrp-3 was dominant in the cerebellar Purkinje cells, the olfactory mitral cells and the striatal large-sized intrinsic neurons. Mtrp-4 mRNA was evident in the olfactory bulb, the septum, the hippocampal neuronal layers, and the cerebellar granule cell layer, while the expression for Mtrp-6 was rather confined to the dentate granule cell layer. Their differential localization suggests that the individual homologues exert their functions in region-specific and neuron specific manners in the calcium signaling. PMID- 9747654 TI - A case of hemolytic uremic syndrome associated with emphysematous cholecystitis and a liver abscess. AB - Hemolytic uremic syndrome (HUS) is characterized by microangiopathic hemolytic anemia, thrombocytopenia and acute renal failure. Most cases of HUS are characterized by a prodromal phase of diarrhea and melena, and affect mainly children. Here we report a unique case of adult-onset HUS that was associated with emphysematous cholecystitis and a liver abscess. The patient did not suffer from diarrhea or melena on admission, but abdominal CT scans revealed emphysematous cholecystitis and a liver abscess. Cholecystectomy was performed and the liver abscess was drained. Cultures of the bile and liver abscess contents were negative, but the serum samples had antibodies against Escherichia coli (E. coli) O157. The patient was anuric for 14 days, and underwent hemodialysis that was repeated 15 times and plasma exchanges 6 times. She recovered from acute renal failure but with inadequate urinary concentrating ability as a sequela. Histopathological examination of renal biopsy specimens on the 83rd hospital day revealed almost normal glomeruli and patchy atrophy of tubules with an increase of interstitium. This is a very rare case of HUS associated with emphysematous cholecystitis and a liver abscess successfully treated with aggressive supportive care. It is possible that an infection with verotoxin-producing E. coli O157 caused the disease. PMID- 9747655 TI - Suppression of gene expression and production of interleukin 13 by dexamethasone in human peripheral blood mononuclear cells. AB - We examined the effect of dexamethasone on the gene expression and production of interleukin (IL)-13 by human peripheral blood mononuclear cells from healthy controls. The gene expression was assessed semiquantitatively by sequential transcription polymerase chain reaction and Southern blot analysis, and the production of this cytokine was assessed by enzyme-linked immunosorbent assay (ELISA). Dexamethasone suppressed IL-13 gene expression induced by stimulation with phytohemagglutinin and phorbol 12-myristate 13-acetate in a dose-dependent manner, with 96% suppression at 10(-6) M, and also suppressed the increased production of IL-13. This is suggested to be one of the mechanisms by which glucocorticoids suppress allergic inflammation. PMID- 9747656 TI - Quinolone photoallergy: photosensitivity dermatitis induced by systemic administration of photohaptenic drugs. AB - Quinolone antibacterial agents are well known to elicit photosensitivity as a side effect. The photoallergenicity of fluoroquinolones, the representative quinolone derivatives, is mainly derived from their photohaptenic moiety. When epidermal cells are irradiated with ultraviolet A light in the presence of fluoroquinolones, quinolone photoadducts are formed in the treated cells. This photomodification is thought to be an initial step for sensitization and elicitation of this photoallergy, and quinolone-photoderivatized Langerhans cells are capable of stimulating immune T cells in mice. In the murine model, fluoroquinolone photoallergy is mediated by Th1 cells bearing T cell receptor Vbeta 13. There is a broad photoantigenic cross-reactivity among fluoroquinolones in recognition by T cells and immunoglobulins. Therefore, it is most likely that fluoroquinolones carry the same photoantigenic epitope, which is recognized by Vbetal3+ T cells, leading to fluoroquinolone photosensitivity and cross reactivity. PMID- 9747657 TI - The photoprotective effect of 1,25-dihydroxyvitamin D3 on ultraviolet light B induced damage in keratinocyte and its mechanism of action. AB - We investigated the photoprotective effect of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) both in vivo and in vitro, revealing its relationship with glutathione, a well-known antioxidant. We also probed into the possible mechanism of photoprotection of 1,25(OH)2D3 through immunohistochemical study for metallothionein (MT). At the same time, endogenous antioxidant effect of 1,25(OH)2D3 was examined. Survival of cultured human keratinocytes was decreased when the cells were irradiated with ultraviolet light-B (UVB) at doses above 30 mJ/cm2. But in the presence of 1,25(OH)2D3 (12 nM), the decrease of survival of keratinocytes by UVB was diminished. The formation of sunburn cells by UVB irradiation in the skin of ICR mice was inhibited by topical application of 1,25(OH)2D3, regardless of prior glutathione depletion. Immunohistochemical staining revealed that 1,25(OH)2D3 induced the expression of MT, a potent radical scavenger, mainly in the basal layer of ICR mice skin. 1,25(OH)2D3 neither inhibited peroxidation of plasma lipids nor interacted with superoxide, nor removed hydrogen peroxide as an antioxidant. These findings suggest that 1,25(OH)2D3 has photoprotective effect not related with glutathione or its endogenous antioxidant property. Rather, it could be attributed to 1,25(OH)2D3 induced MT and its capacity to prevent radical-related damage in UVB irradiation. PMID- 9747658 TI - Sensitivity of cross-reacting antihuman antibodies in formalin-fixed porcine skin: including antibodies to proliferation antigens and cytokeratins with specificity in the skin. AB - Although no animal is a perfect skin model for the study of toxicological and therapeutic agents, structurally the pig may be superior to even non-human primates. Because our work involves effects of toxicological and therapeutic agents on the skin, we wanted to identify stains which may prove useful as well as determine cross-reactivity of some newer antihuman antibodies. We performed a battery of formalin-fixed skin from weanling pigs and minipigs. The battery of antibodies included LCA, CD3, OPD-4, CD34, UCHL-1, L-26, KP-1, MAC-387, Factor XIIIa, Leu-7, S-100 protein, HMB-45, GFAP, synaptophysin, neurofilament protein, ubiquitin, vimentin, type IV collagen, laminin, fibronectin, Factor VIII related antigen, Desmin-M, smooth muscle actin, cytokeratin 7, cytokeratin 20, AEI/AE3, CAM 5.2, EMA, GCDFP, Ki-67, and PCNA. Immunohistochemical stains for CD3, Leu-7, S-100 protein, type IV collagen, laminin, Factor VIII related antigen, GFAP, synaptophysin, neurofilament protein, ubiquitin, smooth muscle actin, vimentin, Desmin-M, cytokeratin 7, cytokeratin 20, AE1/AE3, CAM 5.2, Ki-67 and PCNA showed consistent cross-reactivity. In formalin-fixed tissue, only antibodies to lymphoreticular cells showed poor cross-reactivity. A high percentage of the remaining antibodies did show good cross-reactivity but with some interesting similarities and differences in specificity. PMID- 9747659 TI - Search for human T-lymphotropic virus type I carriers among northeastern Chinese. AB - Human T-lymphotropic virus type I (HTLV-I) is thought to be the causative agent of adult T-cell leukemia/ lymphoma (ATL). This virus infection is endemic in southwestern Japan, parts of Africa and the Caribbean Islands. We examined sera of 1645 subjects of Liaoning province, northeastern China to detect HTLV-I carriers in an effort to reveal the migratory route taken by the early Japanese (Jomon people). As a result, all sera were found to be negative as tested by particle-agglutination (PA), immunofluorescence (IF), enzyme-linked immunoabsorbent (ELISA) and Western blotting methods. This suggests that the Jomon people, who are thought to have brought HTLV-I to the Japan archipelago tens of thousands of years ago, did not come from northeast China. PMID- 9747660 TI - Taurin-conjugated ursodeoxycholic acid has a reversible inhibitory effect on human keratinocyte growth. AB - Tauroursodeoxycholic acid (TUDC) is one of the most hydrophilic taurin conjugated bile acids. TUDC has a suppressive effect on DNA synthesis in primary cultured rat hepatocytes. In this study, we investigated the growth inhibitory effect of TUDC on cultured human keratinocytes. TUDC suppressed the proliferation of keratinocytes in a dose dependent fashion, as measured by both cell counts and 5 bromo-2'-deoxyuridine (BrdU) uptake. Keratinocytes reproliferated and reached almost the same cell number as control after removal of TUDC from the medium. TUDC (1 mM) had no effect on the cell viability, as measured by the dye exclusion test. Epidermal sheets stratified in the presence of TUDC appeared thinner than those stratified without TUDC. These results suggest that TUDC has a reversible growth suppressive effect on human keratinocytes through the mechanism other than cytotoxicity and would be applicable for the treatment of hyperproliferative skin disorders such as psoriasis. PMID- 9747661 TI - Epidermal cell kinetics of pig skin in vivo following UVB irradiation: apoptosis induced by UVB is enhanced in hyperproliferative skin condition. AB - We investigated the effects of ultraviolet B (UVB) irradiation on pig epidermal sunburn cell (apoptotic cell) formation. Expression of p53 tumor suppressor gene product, p21 (WAF1/CIP1), and proliferating cell nuclear antigen (PCNA) was also determined immunohistochemically. Apoptotic cells appeared at 12 h and reached a peak at 48 h following 2 MED-UVB irradiation. The formation of sunburn cells was confirmed by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling (TUNEL) method. p53-positive cells, and p21-positive cells appeared at 6 h, and 12 h, respectively, following the UVB-irradiation. The peak of p53 positive cells was observed at 24 h, and that of p21-positive cells was at 48 h. No expression of TUNEL-, p53-, or p21-positive cells was detected in non irradiated epidermis. The increase in PCNA-positive cells was observed at 24 h and reached its peak at 96 h following the UVB-irradiation. Flow cytometric analyses indicated a decrease in S-phase cells at 24 h, that was followed by their increase at 96 h. Cells in G2/M phase were also considerably decreased at 6 h and 48 h following the UVB-irradiation, and was followed by their increase thereafter. The [3H]thymidine uptake and mitotic counts remained low up until 48 h, and then both parameters increased reaching their peaks at 72 96 h. Effects of UVB irradiation were also determined in tape stripping-induced hyperproliferative epidermis. The numbers of UVB-induced apoptotic cells and PCNA positive cells were markedly enhanced in the tape stripping-treated epidermis, while the numbers of p53- and p21-positive cells were not significantly altered. No induction of apoptosis, p53, or p21 was observed by tape stripping alone. Our results indicate that UVB irradiation induces G1 arrest, prolonged S, and G2/M block of epidermal keratinocytes as well as apoptosis. These processes provide a G1 check point and the elimination of possibly hazardous cells carrying DNA damage, respectively. Our results also indicate that the UVB-induced apoptotic process is enhanced in hyperproliferative skin condition suggesting that apoptosis is closely associated with cell cycle progression. PMID- 9747662 TI - Protein kinase C-alpha levels are inversely associated with growth rate in cultured human dermal fibroblasts. AB - Human dermal fibroblasts are known to express the alpha, delta, epsilon, and zeta isoforms of protein kinase C (PKC). We asked whether the growth of human dermal fibroblasts correlates with expression of a particular PKC isoform. Of total PKC activity measured in the presence of calcium, a condition permissive for activation of all PKC isoforms, 75%) was contributed by PKC-alpha, suggesting that PKC-alpha is the dominant isoform in human dermal fibroblasts. We then further studied PKC-alpha under different culture conditions and in cultures derived from different aged donors. In both subconfluent and confluent cultures, total PKC activity and the level of PKC-alpha protein were consistently higher in slowly proliferating adult cells than in more rapidly proliferating newborn cells. Moreover, in newborn fibroblasts density strongly influenced these parameters. At subconfluent density, when cells were dividing exponentially, total PKC activity was 345+/-63 cpm/,ug protein; whereas at confluent density, when cells were growth arrested, it was 6-7 fold higher, 2334+/-50 cpm/ug protein. Immunoblot analysis using a specific monoclonal antibody against PKC alpha exhibited a similar 6-7 fold increase in the level of PKC-alpha protein at confluent density. However, in adult cells, density had no influence on the already high total activity or level of PKC-alpha. To further determine whether the increases in the levels of total PKC activity and the alpha isoform correlate with the decreased growth rate, a characteristic of both adult donor-derived and confluent cells, total PKC activity and the level of PKC-alpha in subconfluent quiescent cells was compared to that in paired exponentially growing cells at the same density. Total PKC activity was 8836+/-71 cpm/microg protein in subconfluent quiescent cells versus 4415+/-175 cpm/microg protein in dividing cells. The level of PKC-alpha protein was also 2-3 fold higher in quiescent than in growing cultures. However, the amount of PKC-alpha mRNA in these two conditions was identical as determined by northern blot analysis. Taken together, these results suggest an inverse relationship between the levels of total PKC activity and PKC alpha protein and fibroblast growth rate that is regulated at the post transcriptional level. PMID- 9747663 TI - Candida is not involved in the development of of periungual psoriatic lesion. AB - Infection can be a trigger or an aggravating factor in psoriasis, and in particular, bacterial focal infection is well documented to exacerbate psoriasis presumably through its superantigen. Since fungi could produce superantigen as well, and periungual involvement is often seen in psoriatic patients, we examined if local Candida infection plays some pathogenetic role or not in periungual lesions. In 15 psoriatic patients with periungual involvement, Candida infection was examined by microscopic study, culture, and antifungal treatment. We found no evidence suggestive of the relationship between Candida and psoriatic changes in periungual area. PMID- 9747664 TI - Harmonising adverse drug reaction terminology: the role of the Council for International Organizations of Medical Sciences. AB - Health professionals from different countries are known to differ considerably in their use of medical terminology, including the terminology used for adverse drug reactions (ADRs) and in the exact meanings attributed to terms. To remedy this situation, the Council for International Organizations of Medical Sciences (CIOMS) has attempted to provide definitions and basic requirements for proper use of ADR terms. The work has concentrated on terms liable to be misinterpreted and those used for serious and frequently reported ADRs. For every selected term a short monograph has been prepared. It consists of a preamble, definition, basic requirements for use of the term and additional comments, if any. In cooperation with medical experts, drug regulators and the pharmaceutical industry, 13 papers have been published so far. Approximately 160 terms have been defined and work on another 50 terms continues. The full collection of monographs will eventually appear in the form of a book and CD-ROM intended to help doctors fill in case reports, and regulatory agencies and the pharmaceutical industry assess reports. Pharmaceutical companies receive numerous reports of suspected ADRs from medical practitioners and other prescribing professionals. Each company is required to transmit these reports to the drug regulatory agency of the country, or countries, in which the drug is used. Therefore, in addition to receiving the correct name of the ADR, collecting and evaluating centres, regardless of whether they are part of a regulatory agency or a pharmaceutical company, need to be provided with sufficient supporting data to be convinced that what is reported was what was actually observed, and that the ADR term used represents the observed event. PMID- 9747666 TI - Safety and tolerability of moxonidine in the treatment of hypertension. AB - Classical centrally acting antihypertensive agents lower blood pressure by reducing excessive sympathetic tone; however, their clinical use is limited by an adverse effect profile resulting from alpha2-adrenoceptor agonism. Moxonidine is a new centrally acting agent showing selective agonism of imidazoline I1 receptors, but very little alpha2-adrenoceptor agonism. The safety and tolerability of moxonidine was reviewed over an 8-year period (1989 to 1997), including 74 clinical trials and an estimated 370000 patient-years of exposure. Dry mouth and somnolence were the most frequently reported adverse events, followed by headache and dizziness. In phase II to IV controlled studies in patients with hypertension (n = 1460), the incidence of dry mouth was 8 to 9%, somnolence 5 to 8% and headache 6%, as recorded by spontaneous reporting; the percentage of patients discontinuing treatment because of adverse events did not exceed 4%. Subgroup analyses revealed no differences in adverse events related to age or gender. Moxonidine did not exacerbate concomitant conditions such as diabetes mellitus or chronic obstructive pulmonary disease, or interact pharmacokinetically with concurrent medications such as hydrochlorothiazide, digoxin and glibenclamide (glyburide). Coadministration of moxonidine with lorazepam resulted in small additional impairments in tasks requiring attention. A similar distribution of adverse events was observed in uncontrolled studies (n = 1058). The incidence and severity of dry mouth and somnolence were found to decrease with increasing exposure to moxonidine over a period of up to 2 years. Serious adverse events were rare in all trials and could not be attributed to administration of moxonidine. Post-marketing surveillance of the adverse effect profile of moxonidine detected 2 additional adverse effects: nausea and allergic skin reactions. The safety profile of moxonidine, combined with proven antihypertensive efficacy, suggests that it may have an important role to play in the management of mild-to-moderate hypertension. PMID- 9747665 TI - Adverse effects of opioid agonists and agonist-antagonists in anaesthesia. AB - The traditional view of opioids held that the individual opioid agonists shared the same mechanism of action, differing only in their potency and pharmacokinetic properties. However, recent advances in opioid receptor pharmacology have made this view obsolete. Distinguishing features of the synthetic opioid agonists are related, at least in part, to variation in affinity and intrinsic efficacy at multiple opioid receptors. Respiratory depression is the opioid adverse effect most feared by anaesthesiologists. Specific kappa-receptor agonists produce analgesia with little or no respiratory depression. There are a number of commercially available kappa-receptor partial agonist drugs, the so-called agonist-antagonist or nalorphine-like opioids, which appear to have a limited effect on breathing. Within the series of fentanyl analogues there are differences in behaviour towards particular opioid receptors and there is evidence for subtle differences in respiratory depressant effects. Pethidine (meperidine) causes histamine release and myocardial depression, while the fentanyl analogues do not. Pethidine has atropine-like effects on heart rate, while fentanyl analogues reduce heart rate by a vagomimetic action. Severe bradycardia or even asystole is possible with fentanyl analogues, especially in conjunction with the vagal stimulating effects of laryngoscopy. Fentanyl analogues often produce minor reductions in blood pressure, and occasionally severe hypotension by centrally mediated reduction in systemic vascular resistance. Muscle rigidity and myoclonic movement occurs frequently during induction of anaesthesia with larger doses of opioids. Fentanyl and alfentanil have been reported to produce localised temporal lobe electrical seizure activity in patients with complex partial epilepsy. There are probably fewer biliary effects with agonist-antagonist opioids than the agonist opioids. The mechanism of adverse effects after spinal administration is distinctly different for morphine, which is very water soluble, compared with more lipid-soluble opioids. The systemic absorption of morphine after intrathecal or epidural administration is very slow, resulting in long duration of analgesia and low plasma concentrations, while lipid-soluble opioids are rapidly absorbed into the circulation and redistributed to the brain. The serotonin syndrome may result from coadministration of pethidine, dextromethorphan, pentazocine or tramadol with monoamine oxidase inhibitors (MAOIs) or selective serotonin (5 hydroxytryptamine; 5-HT) reuptake inhibitors (SSRIs). There are clinically important interactions between opioids and hypnosedatives, resulting in synergistic effects on sedation, breathing and blood pressure. PMID- 9747667 TI - A risk-benefit assessment of antileukotrienes in asthma. AB - The antileukotriene drugs are the first new therapeutic agents approved for the treatment of asthma in more than 20 years. The currently available compounds are orally active and either prevent the cysteinyl leukotrienes from binding to and activating the cysLT-1 receptor in the lung (leukotriene receptor antagonists) or inhibit leukotriene synthesis (leukotriene synthesis inhibitors). Studies performed in individuals without asthma and patients with asthma reveal that antileukotrienes prevent the bronchoconstriction produced by exercise, cold-air, allergen, aspirin (acetylsalicylic acid) and sulphur dioxide. Except for the setting of aspirin sensitivity where the antileukotrienes are nearly uniformly effective, individual responses to them are variable with complete protection in some, no protection in others and a modest degree of protection in the majority. The antileukotrienes bronchodilate the airways of patients with baseline bronchoconstriction, although usually not as well as beta-agonists. When given for weeks to months they rapidly improve pulmonary function and symptoms in patients with mild-to-moderate asthma, and probably in patients with more severe asthma as well, and these improvements persist for the duration of treatment. Here too, their beneficial effects are variable and not predictable based on clinical criteria. Recent studies suggest they can reduce asthma-induced airway inflammation and are equal or more effective than sodium cromoglycate, but equal or less effective than low-to-moderate dosages of inhaled corticosteroids. Initial experience with the antileukotrienes reveals limited toxicity and what appears to be a favourable therapeutic-to-toxic ratio. However, exposure of more patients with differing characteristics for longer periods of time is needed to substantiate this initial impression. The exact role of the antileukotrienes in the treatment of asthma remains to be determined, as does the relative potency of the various agents. PMID- 9747669 TI - Preclinical development of low toxicity drugs: focus on zanamivir, an anti influenza drug. AB - Developing novel compounds with low toxicity may present more difficulties for pharmaceutical companies than developing compounds with known class-related effects. The absence of clearly identified toxicity may be a consequence either of an inadequate or poorly designed toxicity programme or of the very low toxicity of the novel compound. To enable an informed risk assessment to be undertaken prior to registration, regulatory authorities must satisfy themselves that all efforts to fully evaluate the toxicity profile of a novel compound have been made. Zanamivir is a novel antiviral agent developed for the treatment and prevention of influenza when administered by the oral inhaled route. The toxicology programme for zanamivir was designed to support both a short term treatment indication for patients clinically diagnosed with influenza and a longer term treatment indication for the prevention of influenza. The toxicology studies demonstrated that zanamivir has very low toxicity and no drug-specific toxicities were observed in animal toxicity studies. Systemic plasma concentrations 1336-fold those achieved in clinical use were not associated with significant adverse effects. In the absence of dose-limiting toxicity in animal studies and in an attempt to identify target-organ toxicity, the high dosage level in all repeat dose studies was selected to be the maximum practicable. In the rat, nonspecific effects were seen in the respiratory tract following long term inhaled administration and in the kidneys following continuous infusion. However, these nonspecific effects were consequences of the excessive dosages administered and are not related specifically to zanamivir; thus, they are without relevance to the clinical use of this agent. PMID- 9747668 TI - Drug safety issues in pregnancy following transplantation and immunosuppression: effects and outcomes. AB - Successful pregnancy outcomes are possible after solid organ transplantation. While there are risks to mother and fetus, there has not been an increased incidence of malformations noted in the newborn of the transplant recipient. It is essential that there is closely coordinated care that involves the transplant team and an obstetrician in order to obtain a favourable outcome. Current data from the literature, as well as from reports from the National Transplantation Pregnancy Registry (NTPR), support the concept that immunosuppression be maintained at appropriate levels during pregnancy. At present, most immunosuppressive maintenance regimens include combination therapy, usually cyclosporin or tacrolimus based. Most female transplant recipients will be receiving maintenance therapy prior to and during pregnancy. For some agents, including monoclonal antibodies and mycophenolate mofetil, there is either no animal reproductive information or there are concerns about reproductive safety. The optimal (lowest risk) transplant recipient can be defined by pre-conception criteria which include good transplant graft function, no evidence of rejection, minimum 1 to 2 years post-transplant and no or well controlled hypertension. For these women pregnancy generally proceeds without significant adverse effects on mother and child. It is of note that the epidemiological data available to date on azathioprine-based regimens are favourable in the setting of a category D agent (i.e. one that can cause fetal harm). Thus, there is still much to learn regarding potential toxicities of immunosuppressive agents. The effect of improved immunosuppressive regimens which use newer or more potent (and potentially more toxic) agents will require further study. PMID- 9747670 TI - The protein family of RNA helicases. AB - RNA helicases represent a large family of proteins that have been detected in almost all biological systems where RNA plays a central role. They are ubiquitously distributed over a wide range of organisms and are involved in nuclear and mitochondrial splicing processes, RNA editing, rRNA processing, translation initiation, nuclear mRNA export, and mRNA degradation. RNA helicases are described as essential factors in cell development and differentiation, and some of them play a role in transcription and replication of viral single stranded RNA genomes. Comparisons of the conserved sequences reveal a close relationship between them and suggest that these proteins might be derived from a common ancestor. Biochemical studies have revealed a strong dependence of the unwinding activity on ATP hydrolysis. Although RNA helicase activity has only been demonstrated for a few examples yet, it is generally believed that all members of the largest subgroups, the DEAD and DEAH box proteins, exhibit this activity. PMID- 9747671 TI - Telomere structure, replication and length maintenance. AB - Telomeres are the termini of linear eukaryotic chromosomes consisting of tandem repeats of DNA and proteins that bind to these repeat sequences. Telomeres ensure the complete replication of chromosome ends, impart protection to ends from nucleolytic degradation, end-to-end fusion, and guide the localization of chromosomes within the nucleus. In addition, a combination of genetic, biochemical, and molecular biological approaches have implicated key roles for telomeres in diverse cellular processes such as regulation of gene expression, cell division, cell senescence, and cancer. This review focuses on recent advances in our understanding of the organization of telomeres, telomere replication, proteins that bind telomeric DNA, and the establishment of telomere length equilibrium. PMID- 9747672 TI - Motion automated perimetry identifies early glaucomatous field defects. AB - OBJECTIVE: To determine if motion automated perimetry can identify early glaucomatous visual field defects in patients with suspected glaucoma (by disc), those with ocular hypertension, and those with primary open-angle glaucoma. METHODS: Motion automated perimetry, a foveally centered motion test, and standard visual field tests were conducted on one randomly selected eye of normal patients (n = 38), patients with suspected glaucoma (by disc) (n = 28), patients with ocular hypertension (n = 18), and patients with primary open-angle glaucoma (n = 21). Subjects' performance on both motion tests were compared with their performance on standard perimetry. RESULTS: Perimetric motion thresholds significantly distinguished the groups (P< or =.001), while the foveally centered motion test was unable to separate them (P< or =.32). Of the total patients, 90.5% of those with glaucoma, 39.3% of those with suspected glaucoma, 27.8% of those with ocular hypertension, and 5.3% of the normal subjects had abnormal results on motion automated perimetry testing. Perimetric motion thresholds were significantly correlated with standard visual field thresholds (P< or =.001). CONCLUSION: Motion automated perimetry identifies visual field defects in patients who already show standard visual field loss as well as in a moderate percentage of those with suspected glaucoma and ocular hypertension, indicating that the testing of discrete locations might be necessary for increased diagnostic utility. PMID- 9747673 TI - Foveal outer retinal function in eyes with unexplained visual symptoms or acuity loss. AB - OBJECTIVE: To determine whether foveal outer retinal dysfunction is common in eyes with unexplained visual symptoms or acuity loss. DESIGN: Prospective study. PARTICIPANTS: Seventy-three eyes of 44 consecutive patients with unexplained visual symptoms or acuity loss, 39 eyes of 39 control subjects, and 12 eyes of 7 patients with known maculopathy. INTERVENTION: Foveal cone electroretinography (ERG) and letter recognition perimetry. MAIN OUTCOME MEASURES: Foveal cone ERG data. RESULTS: Abnormal foveal cone ERG data were recorded in 35 (48%) of 73 eyes (23 [52%] of 44 patients). Among these 35 eyes, amplitude was lower than in normal controls (P<.001) and was correlated with visual acuity and the number of letter recognition perimetry errors (P<.05 for both). The latter was higher in eyes with abnormal retinal responses than in symptomatic eyes with normal responses (P<.01). However, initial symptoms, visual acuity, and macular appearance did not differentiate between these 2 groups. Foveal cone ERG test vs retest data showed consistent results. CONCLUSION: Foveal outer retinal dysfunction is a common underlying mechanism of previously unexplained visual symptoms or acuity loss. Foveal cone ERG testing should be considered early in the evaluation of eyes with this presentation. PMID- 9747674 TI - Foveal dysfunction and central visual field loss in glaucoma. AB - OBJECTIVE: To determine whether foveal function distal to the ganglion cell layer is an independent predictor of central visual field function in glaucoma. SETTING: University affiliated hospital and private practice. PARTICIPANTS: Twenty-seven eyes (27 patients) with normal-pressure glaucoma, 10 eyes (10 patients) with primary open-angle glaucoma, and 47 eyes of 47 matched normal volunteers. INTERVENTION AND MAIN OUTCOME MEASURES: Foveal cone electroretinogram (ERG) amplitude, relative optic cup to disc area and their relations to Humphrey full-threshold 30-2 visual field central 4-point mean total deviation (C4MTD) and pattern deviation (C4MPD). RESULTS: Foveal cone ERG amplitude was subnormal in 14 (37.8%) of the 37 glaucomatous eyes and lower in the glaucoma group compared with normal eyes (P<.01). The C4MTD and C4MPD were lower in glaucomatous eyes with subnormal amplitudes compared with those with normal amplitudes (P<.01 and P<.05, respectively). Amplitude was directly correlated with C4MTD (P<.01) and C4MPD (P<.01). Relative optic cup to disc area was inversely correlated with C4MTD (P<.001) and C4MPD (P<.001). Partial correlation analysis revealed that amplitude and relative optic cup to disc area were independent predictors of C4MTD and C4MPD. CONCLUSION: Foveal function distal to the ganglion cell layer and optic disc cupping independently predict central visual field function in glaucoma. PMID- 9747675 TI - Foveal cysts: a premacular hole condition associated with vitreous traction. AB - OBJECTIVE: To define the clinical findings, cause, and outcome of patients with foveal cysts due to vitreous traction. METHODS: Follow-up of 18 patients with foveal cysts and no posterior vitreous detachment (PVD). Changes were documented in visual acuity, the appearance of the fovea, or the development of a macular hole or PVD. We studied 8 eyes using the retinal thickness analyzer. RESULTS: On follow-up, 9 of 23 eyes did not develop a PVD and still had a foveal cyst; 8 of 23 developed a full-thickness macular hole; 4 of 23 developed a PVD with resolution of the cyst; and 2 eyes underwent vitrectomy for the cyst before a full-thickness hole developed. Analysis with the retinal thickness analyzer showed splitting within the middle retinal layers and in some cases unroofing or absent inner retinal layers in the center of the cyst. CONCLUSIONS: Foveal cysts are caused by vitreous traction. These eyes may remain stable, develop full thickness holes, or develop a PVD with resolution of the cystic changes. A foveal cyst seems to be a common finding in patients with foveal traction from a variety of mechanisms. PMID- 9747676 TI - Inherited retinal arteriolar tortuosity with retinal hemorrhages. AB - BACKGROUND: Familial arteriolar tortuosity is an autosomal dominant disorder affecting the retinal arterioles. OBJECTIVES: To report a pedigree with this disorder and describe a systemic workup to determine whether this vascular abnormality is limited to the eye. RESULTS: A 58-year-old woman referred for retinal hemorrhages was found to have retinal arteriolar tortuosity of both eyes, especially in the macular area. Her 63-year-old brother had a history of retinal hemmorhages beginning at age 18 years and had similar fundoscopic examination findings. The proband had an extensive systemic workup, including magnetic resonance imaging, and cardiac and renal angiography, that failed to demonstrate any other sequelae of this inherited ocular syndrome. However, each member of the family expressing this phenotype did have hypertension. CONCLUSION: Inherited retinal arteriolar tortuosity is an autosomal dominant disorder limited to the eye, at least in this pedigree, within the sensitivity of the systemic workup we used. PMID- 9747677 TI - Visual outcomes following lensectomy and vitrectomy for combined anterior and posterior persistent hyperplastic primary vitreous. AB - OBJECTIVE: To determine the visual outcome after surgery for persistent hyperplastic primary vitreous using modern vitreoretinal techniques. DESIGN: Retrospective medical record review during a 5-year period (June 1992 to June 1997). Information recorded for each patient included age, medical history, sex, results of preoperative ocular examination, age at diagnosis, procedure performed, intraoperative and postoperative complications, location and number of sclerotomy sites, type of aphakic rehabilitation, amblyopic therapy given, final visual acuity, and length of follow-up. RESULTS: Fourteen patients who underwent surgical management of combined anterior and posterior persistent hyperplastic primary vitreous were identified. Eleven patients underwent aphakic rehabilitation and aggressive amblyopic therapy consisting of occlusive therapy for several waking hours each day. One additional older patient received aphakic rehabilitation only. Ten eyes (71%) achieved a visual acuity of 20/300 or better, and 8 (57%) obtained a final visual acuity of 20/100 or better. Average length of follow-up was 22 months (range, 4-57 months). Nine patients were fitted with an aphakic soft contact lens, 2 older patients had a posterior chamber intraocular lens placed at the time of vitrectomy, and 1 patient wore aphakic spectacles. CONCLUSIONS: With modern vitreoretinal techniques, aphakic rehabilitation, and aggressive amblyopic therapy, useful vision can be obtained in the majority of patients with combined anterior and posterior persistent hyperplastic primary vitreous. PMID- 9747678 TI - Disinfection of eyelid speculums for retinopathy of prematurity examination. AB - OBJECTIVE: To evaluate the effectiveness of 70% isopropyl alcohol swabs in disinfecting eyelid speculums after examination for retinopathy of prematurity. METHODS: Two phases. Phase 1: 46 autoclave-sterilized eyelid speculums randomized into either a cleaned or control group following examination for retinopathy of prematurity. Speculums in the cleaned group were disinfected with a 70% isopropyl alcohol swab while control speculums were not cleaned. Bacterial and fungal cultures were then obtained. Phase 2: 20 autoclave-sterilized eyelid speculums inoculated with a clinically relevant dilution of adenovirus serotype 5 or herpes simplex virus type 2. Inoculated speculums were randomized into either a cleaned or control group. RESULTS: Phase 1: 17 (70.8%) of 24 cultures from the cleaned group yielded bacteria compared with 21 (95.5%) of 22 controls. Fungi were isolated from only 1 control and from no cleaned speculums. Phase 2: all speculums inoculated with adenovirus supported growth of the organism irrespective of cleaning with 70% isopropyl alcohol swabs. None of 5 cleaned speculums inoculated with herpes simplex virus type 2 supported viral growth, compared with 3 (60%) of 5 cultures positive for growth in the control group. CONCLUSION: Cleaning eyelid speculums with 70% isopropyl alcohol swabs provided inadequate disinfection against bacteria following examination for retinopathy of prematurity and against adenovirus in a laboratory simulation. PMID- 9747680 TI - Gelatinase B and A expression after laser in situ keratomileusis and photorefractive keratectomy. AB - OBJECTIVE: To compare the expression of gelatinases in the corneal epithelium and stroma after laser in situ keratomileusis (LASIK) and photorefractive keratectomy (PRK). METHODS: Rabbit eyes were treated with LASIK (n=11), PRK (n=12), or corneal flap construction (n=12); 4 eyes served as unwounded controls. Zymography was performed on the central epithelium and the stroma 1, 3, and 7 days after surgery to determine the expression of gelatinases. RESULTS: Epithelial expression of gelatinase B in the LASIK group (0%-25%) was lower than that in the PRK group at all time points (50%-100%) and was identical to the corneal flap group. Stromal expression of gelatinases A and B was similar after LASIK and PRK, but was minimal after corneal flap construction at all time points. Epithelial expression of gelatinase A was similar for the first 3 days after LASIK and PRK but not thereafter. CONCLUSIONS: Gelatinase B epithelial expression was up regulated after PRK but not after LASIK. Gelatinase B stromal expression was up regulated after both procedures. CLINICAL RELEVANCE: Differences in wound healing and subepithelial scarring after these 2 procedures may be related to gelatinase B. PMID- 9747679 TI - Intraorbital implants after enucleation and their complications: a 10-year review. AB - BACKGROUND: Many different types of orbital implants have been used after enucleation. Associated complications such as infection, exposure, extrusion, and ptosis have been reported. OBJECTIVE: To describe 342 consecutive patients who underwent enucleation with intraorbital implant placement at the Bascom Palmer Eye Institute, University of Miami School of Medicine, Miami, Fla, during the past 10.5 years and their complications. METHODS: Medical records of orbital implantation after enucleation performed by 3 surgeons (T.G.M., D.T., and T.J.) were reviewed retrospectively. Demographic data, ocular diagnosis, previous ophthalmic surgery, implant characteristics, and postoperative complications were described in all patients, with a minimum of 2 months' follow-up, using a standardized format. RESULTS: Eleven complications were observed in 7 patients. Four patients had exposure of the implant, and 1 of these patients developed associated infection. Three patients developed pyogenic granulomas, 1 patient developed ptosis requiring surgical intervention, 1 patient had long-term orbital discomfort, and 1 patient developed an inclusion cyst. CONCLUSION: Complications after enucleation with orbital implant placement are minimal and are observed with both porous and acrylic orbital implants. PMID- 9747681 TI - Intraocular concentrations of chemotherapeutic agents after systemic or local administration. AB - OBJECTIVES: To investigate the concentrations of carboplatin and etoposide achieved in the aqueous and vitreous humors after intravenous infusion in nonhuman primates, and to investigate whether local administration of carboplatin might result in higher concentrations in the vitreous humor. METHODS: Macaca fascicularis primates were treated with 1 of 3 regimens: (1) intravenous carboplatin (18.7 mg/kg), etoposide (5 mg/kg), and vincristine sulfate (0.05 mg/kg), (2) peribulbar carboplatin (10 mg/mL), or (3) episcleral balloon carboplatin (10 mg/mL). Concentrations of chemotherapeutic agents were measured in the plasma and in the aqueous and vitreous humors. RESULTS: No measurable amount of etoposide was detected in the aqueous or vitreous humor after intravenous administration. Mean measured peak vitreous concentration of carboplatin after intravenous administration was 0.31 microg/mL, which was 1% of the peak plasma value. Mean measured peak vitreous concentrations of carboplatin after peribulbar or episcleral balloon administration were 2.38 microg/mL and 2.95 microg/mL, respectively, which represent 7.68- and 9.52-fold increases over the concentration achieved after intravenous administration. No serious toxic effect was observed in any animal. CONCLUSIONS: Peribulbar and episcleral balloon administration of carboplatin seemed to be safe and resulted in higher vitreous concentrations than intravenous administration in this model. These results suggest that these alternate routes of delivery should be explored in children with vitreous seeding of retinoblastoma. PMID- 9747682 TI - Effect of 8-iso prostaglandin E2 on aqueous humor dynamics in monkeys. AB - OBJECTIVE: To evaluate the effects of 8-iso prostaglandin E2 (8-iso PGE2; prosta 5,13-dien-1-oic acid,11,15-dihydroxy-9-oxo-,[5Z,8beta-11X,13E,15 S]-) on the intraocular pressure (IOP), outflow facility, and aqueous humor flow rates in normal monkeys and monkeys with glaucoma. METHODS: The IOP was measured before and as long as 6 hours after the topical application of 8-iso PGE2 to 1 eye of 6 normal monkeys and to the glaucomatous eye of 8 monkeys with unilateral laser induced glaucoma. The pupil diameter was measured at the same times as the IOP measurements in the normal monkeys. Tonographic outflow facility and fluorophotometric flow rates of aqueous humor were measured in 6 normal monkeys before and after drug treatment. RESULTS: In normal monkeys, a single dose of 0.1% 8-iso PGE2 reduced (P<.01) the IOP for 4 hours in the treated eyes with a maximum (mean +/- SEM) reduction of 3.2 +/- 0.2 mm Hg, compared with the contralateral control eyes. The pupil size was smaller (P<.01) in the treated eyes by as much as 1.0 +/- 0.2 mm for 4 hours. In 8 glaucomatous monkey eyes, the application of 0.05% and 0.1% 8-iso PGE2 reduced the IOP (P<.01) for as long as 2 and 5 hours, respectively. The maximum reduction in the IOP was 4.6 +/- 0.8 mm Hg (0.05%) and 6.0 +/- 0.8 mm Hg (0.1%) compared with baseline measurements. The magnitude and duration of the ocular hypotensive effect were enhanced with twice a-day administration for 5 consecutive days. Outflow facility in normal monkey eyes was increased (P<.05) by 48% in the treated eyes, and aqueous humor flow was unchanged (P>.10), compared with vehicle-treated contralateral control eyes. Mild eyelid edema, conjunctival edema, hyperemia, and discharge appeared in some eyes treated with the 0.1% drug concentration. CONCLUSIONS: The use of 8-iso PGE2 reduces the IOP in both normal and glaucomatous monkey eyes. An increase in outflow facility appears to account for most of the IOP reduction in normal monkeys. CLINICAL RELEVANCE: The application of 8-iso PGE2 may have potential for the treatment of glaucoma as an outflow facility-increasing drug. PMID- 9747683 TI - Efficacy of hyaluronidase in reducing increases in intraocular pressure related to the use of viscoelastic substances. AB - OBJECTIVE: To evaluate the efficacy of hyaluronidase in preventing increases in intraocular pressure related to injections of hyaluronan-containing viscoelastic substances. METHODS: Twenty-five white rabbits were divided into 5 groups. In groups 1 through 4, 0.15 mL of aqueous humor was removed and replaced with 0.10 mL of a viscoelastic substance in both eyes. Additionally, 10 units of hyaluronidase (0.05 mL) was injected in the anterior chamber of the right eye, whereas the left eye was injected with a volumetrically equivalent dose of balanced saline solution. Viscoelastic substances tested were Healon and Healon GV (Pharmacia & Upjohn, Kalamazoo, Mich), Viscoat (Alcon Laboratories, Fort Worth, Tex), and Ocucoat (Storz Ophthalmics, Clearwater, Fla). In group 5, right eyes were injected with 10 units of hyaluronidase and the left eyes were treated with balanced saline solution. RESULTS: After injections of viscoelastic substance, intraocular pressure rose rapidly, reaching a peak at approximately 46 hours after injection and returning to preinjection levels within 24 hours. Hyaluronidase significantly decreased intraocular pressure when used with Healon, Healon GV, and Viscoat, but not with Ocucoat. When injected in the absence of viscoelastic, hyaluronidase appeared to decrease intraocular pressure, but this result was not statistically significant. CONCLUSIONS: Injections of hyaluronidase into the anterior chamber of rabbits effectively prevent increases in intraocular pressure induced by hyaluronan-containing viscoelastic substances. This effect may be related to the ability of hyaluronidase to cleave hyaluronan moieties. PMID- 9747684 TI - Patterns of emergency department visits for disorders of the eye and ocular adnexa. AB - OBJECTIVE: To characterize the magnitude and patterns of visits to the emergency department (ED) for problems related to the eye and ocular adnexa. METHODS: The National Hospital Ambulatory Medical Care Survey was used to obtain information on ED visits in the United States for conditions of the eye and ocular adnexa in 1993. Patients were identified by International Classification of Diseases, Ninth Revision, Clinical Modification, codes. National projections were based on a staged probability design. RESULTS: There were 2.32 million projected ED visits for problems of the eye and ocular adnexa in 1993. Forty-nine percent of visits were for injuries, two thirds of which occurred in males. Thirty-five percent of injuries occurred in the home and 18% occurred in the workplace. Only 3% of patients required hospitalization. Most patients had private insurance, but substantial variations in coverage existed for patients who used the ED for injury- vs non-injury-related care. CONCLUSIONS: Emergency departments in the United States provide a large amount of eye care, much of which is for conditions other than trauma. Differences in insurance coverage for injury- and non-injury related eye care indicate that factors other than medical urgency are involved in the decision to use ED services. Further studies are needed to determine the cost effectiveness and quality of ocular-related ED visits. PMID- 9747685 TI - Celebrating women in ophthalmology. PMID- 9747686 TI - Assessment of optic disc topography for diagnosing and monitoring glaucoma. PMID- 9747687 TI - Ocular manifestations of Whipple disease: an atypical presentation. AB - A 62-year-old man developed bilateral granulomatous iridocyclitis after uncomplicated cataract surgery. On ophthalmic examination, we found moderate inflammation in the anterior chamber and vitreous, with granular crystalline deposits on the iris, intraocular lens, and capsular bag. Biopsy of the lens capsule and vitreous revealed periodic acid-Schiff-positive, diastase-resistant bacilli consistent with Tropheryma whippelii. Electron microscopy and polymerase chain reaction confirmed the diagnosis of Whipple disease. A jejunal biopsy specimen also revealed T whippelii. Treatment with trimethoprim-sulfamethoxazole, cefixime, rifampin, and doxycycline resulted in improvement of systemic symptoms, but intraocular inflammation persisted. Intraocular inflammation was eventually reduced with the intravenous administration of ceftriaxone sodium. PMID- 9747689 TI - Coexistence of 3 tumors of neural crest origin: neurofibroma, meningioma, and uveal malignant melanoma. AB - OBJECTIVE: To describe the clinical findings in a patient who developed a neurofibroma, meningioma, and choroidal melanoma. METHODS: Clinical and histopathological findings of the case are reviewed and presented. RESULTS: The patient had a right superolateral periorbital neurofibroma, a right sphenoid wing meningioma, and a left choroidal juxtapapillary malignant melanoma. All 3 tumors are derived from neural crest cells. CONCLUSIONS: To our knowledge, this is the first report of a patient with this combination of 3 neural crest-derived tumors. This case is most appropriately classified as a complex neurocristopathy, a disorder involving the aberrant and pathological proliferation of multiple tissues derived from neural crest cells. PMID- 9747688 TI - Parry-Romberg syndrome associated with intracranial vascular malformations. AB - We describe a 23-year-old woman with iridocyclitis, enophthalmos, facial hemiatrophy, and transient numbness of her contralateral upper and lower extremities. The patient was found to have white matter densities in the right hemisphere in magnetic resonance T2-weighted images and vascular malformations involving right vertebral, right carotid, and right anterior cerebral arteries. Histopathologic evaluation of a biopsy specimen of anterior orbital fat and lacrimal gland revealed fibrosis and chronic inflammation. These findings were consistent with the diagnosis of progressive facial hemiatrophy (Parry-Romberg syndrome) in association with iridocyclitis and intracranial vascular malformations. PMID- 9747690 TI - Bilateral ptosis and lower eyelid ectropion secondary to cutaneous leishmaniasis. AB - A 73-year-old white woman had a 14-month history of an extensive, disfiguring facial lesion involving the cheeks, nose, and eyelids, resulting in exposure keratopathy. A biopsy of the facial lesion established the diagnosis of cutaneous leishmania, and the lesion responded to treatment with itraconazole. PMID- 9747692 TI - Hemifacial atrophy and primary corneal endothelial failure. PMID- 9747691 TI - Inadvertent instillation of nonophthalmic antiseptic drops due to packaging similarity. PMID- 9747693 TI - Chronic angle-closure mimicking rubeotic glaucoma in an adult with retinopathy of prematurity. PMID- 9747694 TI - Progressive outer retinal necrosis in a patient with rheumatoid arthritis. PMID- 9747695 TI - Cat-scratch disease manifesting as unifocal helioid choroiditis. PMID- 9747696 TI - Scleral buckle infection with ciprofloxacin-resistant Pseudomonas aeruginosa. PMID- 9747697 TI - Positive temporal artery biopsy 6 months after prednisone treatment. PMID- 9747698 TI - Subepidermal calcified nodule. PMID- 9747699 TI - Free-floating cyst in the anterior chamber. PMID- 9747700 TI - Intralenticular hemorrhage following blunt ocular trauma. PMID- 9747701 TI - Capillary nonperfusion of the retina in diabetes mellitus. PMID- 9747702 TI - Routine antibiotic sensitivity testing for corneal ulcers. PMID- 9747703 TI - Central corneal thickness. PMID- 9747704 TI - Mapping of 5-methylcytosine residues in Nicotiana tabacum 5S rRNA genes by genomic sequencing. AB - Genomic sequencing was used to localise 5-methylcytosine residues in individual DNA strands of 5S rRNA genes in tobacco. The density of methylation along the sequence was high in both strands, exceeding the average methylation density of the tobacco genome. Besides methylation of CG and CNG sequences, considerable amounts of mC were found in non-symmetrical sites. Among 69 sequenced clones obtained from leaf DNA we did not detect any non-methylated clone, and Southern blot hybridisation analysis also failed to suggest the presence of methylation free 5S rDNA units in the tobacco genome. Differences were observed among methylation patterns of individual sequenced clones. This heterogeneity reflects either heterogeneity among individual members of 5S rRNA gene cluster or differences among individual cells. Methylation of CNG and non-symmetrical sites can be efficiently reduced by treatment with dihydroxypropyladenine, an inhibitor of S-adenosylhomocysteine hydrolase. PMID- 9747706 TI - Linkage analysis in tetraploid potato and association of markers with quantitative resistance to late blight (Phytophthora infestans). AB - We have constructed a partial linkage map in tetraploid potato which integrates simplex, duplex and double-simplex AFLP markers. The map consists of 231 maternal and 106 paternal markers with total map lengths of 990.9 cM and 484.6 cM. The longer of the two cumulative map lengths represents approximately 25% coverage of the genome. In tetraploids, much of the polymorphism between parental clones is masked by 'dosage' which significantly reduces the number of individual markers that can be scored in a population. Consequently, the major advantage of using AFLPs--their high multiplex ratio--is reduced to the point where the use of alternative multi-allelic marker types would be significantly more efficient. The segregation data and map information have been used in a QTL analysis of late blight resistance, and a multi-allelic locus at the proximal end of chromosome VIII has been identified which contributes significantly to the expression of resistance. No late blight resistance genes or QTLs have previously been mapped to this location. PMID- 9747705 TI - The chromatin structure of the GAL1 promoter forms independently of Reb1p in Saccharomyces cerevisiae. AB - Positive and negative regulation of the GAL1 promoter of the yeast Saccharomyces cerevisiae results from a network of interactions between transcription factors and chromatin. In this study we used footprinting procedures to characterize these interactions in vivo. DNase I analysis of the GAL1 upstream activating sequence (UAS(GAL1/10)) showed expected Gal4 activator protein binding during growth in galactose, and also revealed binding of the Reb1 protein (Reb1p) during growth in glucose. In addition, we mapped to nucleotide resolution a positioned nucleosome that, in the inactive promoter, packages DNA between the UAS(GAL1/10) and the GAL1 TATA sequence, leaving both of these elements nucleosome free. The nucleosome footprint was lost when the promoter was activated. Surprisingly, mutation of the Reb1p binding site had no effect on nucleosome positioning or on the kinetics or extent of activation or repression of either the GAL1 or GAL10 promoters under any of the conditions assayed. PMID- 9747707 TI - Exopolysaccharide (EPS) synthesis in Bradyrhizobium japonicum: sequence, operon structure and mutational analysis of an exo gene cluster. AB - The nucleotide sequence of a 8330-bp DNA fragment from Bradyrhizobium japonicum 110spc4 was determined. Sequence analysis revealed that six ORFs were present and the deduced amino acid sequences were homologous to enzymes involved in exopolysaccharide (EPS) biosynthesis. The genes appear to be organized into at least four different operons. One gene was found to be homologous to exoB, which encodes a UDP-galactose 4'-epimerase. Other ORFs were homologous to UDP-hexose transferases and one ORF showed similarity to Sinorhizobium (Rhizobium) meliloti ExoP, which has been suggested to be involved in EPS chain-length determination. A set of deletion and insertion mutants was constructed and the resulting B. japonicum strains were tested for their symbiotic traits. Deletion mutant deltaP22, which lacks the C-terminal part of ExoP, the UDP-hexose transferase ExoT and the N-terminal part of ExoB, shows a delayed nodulation phenotype and induces symptoms of plant defense reactions; its EPS does not contain galactose and no high molecular weight fraction is synthesized. In contrast, insertion mutant EH3, which expresses an exoP gene product that is truncated in its putative periplasmic domain, produced an EPS containing both HMW and LMW fractions. However, the interaction of EH3 with soybeans was severely perturbed. As a rule, only the initial steps of nodule formation were observed. PMID- 9747708 TI - The gene for indole-3-acetyl-L-aspartic acid hydrolase from Enterobacter agglomerans: molecular cloning, nucleotide sequence, and expression in Escherichia coli. AB - A 5.5-kb DNA fragment containing the indole-3-acetyl-aspartic acid (IAA-asp) hydrolase gene (iaaspH) was isolated from Enterobacter agglomerans strain GK12 using a hybridization probe based on the N-terminal amino acid sequence of the protein. The DNA sequence of a 2.4-kb region of this fragment was determined and revealed a 1311-nucleotide ORF large enough to encode the 45-kDa IAA-asp hydrolase. A 1.5-kb DNA fragment containing iaaspH was subcloned into the Escherichia coli expression plasmid pTTQ8 to yield plasmid pJCC2. Extracts of IPTG-induced E. coli cultures containing the pJCC2 recombinant plasmid showed IAA asp hydrolase levels 5 to 10-fold higher than those in E. agglomerans extracts. Homology searches revealed that the IAA-asp hydrolase was similar to a variety of amidohydrolases. In addition, IAA-asp hydrolase showed 70% sequence identity to a putative thermostable carboxypeptidase of E. coli. PMID- 9747709 TI - Cloning and characterization of the dnaK heat shock operon of the marine bacterium Vibrio harveyi. AB - We cloned the DNA region of the Vibrio harveyi chromosome containing the heat shock genes dnaK and dnaJ and sequenced them. These genes are arranged in the chromosome in the order dnaK-dnaJ, as in other proteobacteria of the alpha and gamma subdivisions. The dnaK gene is 1923 nucleotides in length and codes for a protein of 640 amino acid residues, with a predicted molecular mass of 69,076 Da and 81.2% similarity to the DnaK protein of Escherichia coli. The V. harveyi dnaJ gene has a coding sequence of 1158 nucleotides. The predicted DnaJ protein contains 385 amino acids, its calculated molecular mass is 41,619 Da and it has 74.7% similarity to the DnaJ protein of E. coli. Northern hybridization experiments with RNA from V. harveyi cells and a DNA probe carrying both the dnaK and dnaJ genes showed a single, heat-inducible transcript, indicating that these genes form an operon. Primer extension analysis revealed five heat-inducible transcriptional start sites upstream of the dnaK gene, two of which (T1 and T4) are preceded by sequences typical of the E. coli heat shock promoters recognized by the sigma 32 (sigma32) factor. Location of these promoters is highly similar to that of the E. coli dnaK promoters. No transcriptional start sites were detected upstream of the dnaJ gene. The V. harveyi dnaKJ operon cloned in a plasmid in E. coli cells was transcribed in a sigma32 dependent manner and the size of the transcript, the kinetics of transcription, and the transcriptional start sites were as in V. harveyi cells. This indicates a high conservation of the transcriptional heat shock regulatory elements between E. coli and V. harveyi, both belonging to the gamma subdivision of proteobacteria. We tested the ability of the cloned dnaKJ genes to complement E. coli dnaK and dnaJ mutants and found that V. harveyi DnaJ restored a thermoresistant phenotype to dnaJ mutants and enabled lambda phage to grow in the mutant cells. V. harveyi DnaK did not suppress the thermosensitivity of dnaK mutants but complemented the dnaK deletion mutant with respect to growth of lambda phage. V. harveyi DnaK, in contrast to DnaJ, failed to modulate the heat shock response in E. coli. Our results suggest that the DnaK chaperone may be more species specific than the DnaJ chaperone. PMID- 9747710 TI - Paracentromeric sequences on tomato chromosome 6 show homology to human satellite III and to the mammalian CENP-B binding box. AB - In order to isolate centromeric sequences from tomato (Lycopersicon esculentum) chromosome 6, a large-scale RAPD screen was performed. Among 1500 polymorphic RAPD markers tested, 100 were identified as chromosome 6-specific, using a L. esculentum substitution line carrying chromosome 6 from L. pennellii. Fifty-seven of these markers proved to originate from L. pennellii, and of these, 13 were genetically mapped between the morphological markers yellow virescent (yv) and thiaminless (tl), which flank the centromere. These markers could be assigned to three genetic loci, with 11 of the markers mapping to a single locus. Further resolution of this cluster was achieved using radiation-induced deletions that removed yv or tl but not the centromere. Seven markers were shown to be located outside all of the deletions. These seven markers and three of the other markers of the cluster were cloned and sequenced. Five of the clones are present as single-copy or low-copy-number sequences and five represent middle repetitive sequences. Three sequences show homology to the mammalian CENP-B binding box; clone AG12 contains two of these boxes and also shows homology to human satellite III. PMID- 9747711 TI - Role of multiple trans-acting regulators in modifying the effect of the retrotransposon copia on host gene expression in Drosophila. AB - We have examined the combinatorial effect of five trans-acting modifiers of white apricot, a copia retrotransposon-induced allele of the white eye-color gene. Quantitation of steady-state levels of copia transcripts for various modifier combinations at different developmental stages using northern analysis reveals nonadditivity and epistasis. Interestingly, extensive overexpression of copia transcripts caused by the modifier Ufo is altered up to twofold in level when Ufo is combined with either of the modifiers Mow or Wow. In general, double combinations do not elevate the level of copia transcripts more than twofold. The role of suppressor mutations in modifying the effect of transposable elements on host gene expression is discussed. PMID- 9747712 TI - Cytosine methylation at CG and CNG sites is not a prerequisite for the initiation of transcriptional gene silencing in plants, but it is required for its maintenance. AB - Transgenes integrated into plant chromosomes, and/or endogenous plant genes, may be subjected to epigenetic silencing at the transcriptional or post transcriptional level. Transcriptional inactivation is correlated with hypermethylation of CG/CNG sites at the silent loci. It is not known whether local hypermethylation is part of the inactivation process, or just an outcome of the silent state. To address this issue, we generated transgenic tobacco lines containing a selectable marker gene controlled by a derivative of the 35S promoter of the cauliflower mosaic virus (CaMV) devoid of CG and CNG methylation acceptor sites. Silencing was triggered by crossing to the silencer locus of tobacco line 271. This line contains inactive and methylated copies of the 35S promoter and is able to silence homologous promoter copies at ectopic chromosomal positions. The mutated promoter lacking CG/CNG methylation acceptor sites was as susceptible to Trans-silencing as the unmodified 35S promoter control. Thus, methylation at CG and CNG sites is not a prerequisite for the initiation of epigenetic gene inactivation. Interestingly, while methylation of the remaining cytosines is usually only slightly affected by silencing, it was significantly increased in the absence of CG/CNG sequences. Since this sequence preference is the same as that of known methyltransferases, this may imply that silencing is accompanied or directly followed by recruitment of methyltransferase, which, in the absence of cytosines in the optimal sequence context, modifies other C residues in the affected area. However, silencing without CG/CNG methylation was immediately relieved in the absence of the silencer. Thus, CG/CNG methylation is probably essential for the maintenance of previously established epigenetic states. PMID- 9747713 TI - Molecular cloning and characterization of the gene encoding N'-[(5' phosphoribosyl)-formimino]-5-aminoimidazole-4-carboxamide ribonucleotide (BBM II) isomerase from Arabidopsis thaliana. AB - We have isolated an Arabidopsis BBM II isomerase cDNA from an Arabidopsis cDNA library, by means of functional complementation of the E. coli hisA mutant strain HfrG6. The isolated cDNA encodes a polypeptide of 304 amino acids with a calculated molecular weight of 33,363. Sequence comparison with the HIS6 proteins of yeasts revealed that Arabidopsis BBM II isomerase contains an N-terminal extension of approximately 40 amino acids that shows the general properties of chloroplast transit peptides. This finding is consistent with the localization of other histidine biosynthetic enzymes, such as imidazoleglycerolphosphate dehydratase and histidinol dehydrogenase, in the chloroplasts in higher plants. The primary structure of the mature protein was 50% and 42% identical, respectively, to the HIS6 proteins of Schizosaccharomyces pombe and Saccharomyces cerevisiae, respectively, while no prominent sequence similarity to the bacterial BBM II isomerase was found. That the isolated Arabidopsis cDNA actually encodes a functionally active BBM II isomerase activity was confirmed in an in vitro enzyme assay using a crude extract prepared from strain HfrG6 transformed with the Arabidopsis BBM II isomerase cDNA. PMID- 9747714 TI - Identification of the DNA-binding sites for two response regulators involved in control of bacteriocin synthesis in Lactobacillus plantarum C11. AB - In Lactobacillus plantarum C11, bacteriocin production has previously been shown to be an inducible process, in which a secreted peptide, produced by the host itself, is involved. The inducing factor, designated plantaricin A (PlnA), is a bacteriocin-like peptide encoded by a gene (plnA) located on the same operon as the genes for a two-component regulatory system (plnBCD). This system consists of a histidine kinase (PlnB) and two response regulators (PlnC,D), and belongs to a recently defined subfamily of two-component regulatory systems, which are activated by secreted peptide pheromones through a quorum-sensing mechanism. We show here that the two response regulators PlnC and PlnD bind specifically to imperfect direct repeats found within the adjacent promoter of the plnABCD operon, and to similar sequences found within the promoter regions of two nearby operons containing bacteriocin structural genes (plnEFI and plnJKLR). Binding of PlnC and PlnD was increased two to three fold in the presence of acetyl phosphate. The results suggest that bacteriocin synthesis in L. plantarum C11 is regulated by the DNA-binding activity of the two response regulators PlnC and PlnD. PMID- 9747715 TI - Herpesviral thymidine kinases: laxity and resistance by design. PMID- 9747716 TI - Analysis of the basal and inducible activities of the ICPO promoter of herpes simplex virus type 1. AB - Sequences from -420 to -70 from the ICPO transcriptional start site of herpes simplex virus type 1 are dispensable for reactivation from latency. A putative cAMP-response element (CRE) outside of this region was non-functional in both murine neuroblastoma (NB41A3) and rat pheochromocytoma (PC12) cells. Also, poor binding of cAMP-response element binding protein (CREB) was observed. Sequences from -95 to -37 are important for constitutive activity in NB41A3, PC12 and baby hamster kidney (BHK) cells. The TATA box and Sp1 site were also shown to be major contributors to constitutive activity. Finally, high constitutive activity of a deleted construct (-420 to -1) in NB41A3 and BHK cells suggests transcription initiates upstream of -420 in the absence of VP16. The implications of these observations regarding ICPO expression during the virus life-cycle are discussed. PMID- 9747717 TI - Reconstitution of CD8 T cells is essential for the prevention of multiple-organ cytomegalovirus histopathology after bone marrow transplantation. AB - Cytomegalovirus (CMV) infection in the period of temporary immunodeficiency after haematoablative treatment and bone marrow transplantation (BMT) is associated with a risk of graft failure and multiple-organ CMV disease. The efficacy of immune system reconstitution is decisive for the prevention of CMV pathogenesis after BMT. Previous data in murine model systems have documented a redundancy in the immune effector mechanisms controlling CMV. CD8 T cells proved to be relevant but not irreplaceable as antiviral effectors. Specifically, in a state of long term in vivo depletion of the CD8 T-cell subset, CD4 T cells were educed to become deputy effectors controlling CMV by a mechanism involving antiviral cytokines. It is of medical importance to know whether one can trust in this 'flexible defence' in all clinical settings. It is demonstrated here that reconstitution of CD8 T cells is crucial for the prevention of fatal multiple organ CMV disease under the specific conditions of BMT. PMID- 9747718 TI - Functional regions of the human cytomegalovirus protein pUL97 involved in nuclear localization and phosphorylation of ganciclovir and pUL97 itself. AB - In order to identify functional regions of the human cytomegalovirus protein pUL97 (i) different 5' fragments of the UL97 open reading frame (ORF) were fused to the coding region of the green fluorescent protein and (ii) recombinant vaccinia viruses (rVV) were generated carrying two full-length and 11 mutated UL97 ORFs. The results indicated the presence of an N-terminal region within pUL97 which changed the intracellular distribution of the fusion proteins. pUL97 was localized in the nucleus, but not in the nucleoli, and was detected in the nuclear matrix fraction. Expression of all pUL97 mutants could be confirmed by Western blot analysis. pUL97-associated ganciclovir (GCV) phosphorylation in rVV infected cells, determined quantitatively by HPLC analysis, was abolished completely using individual UL97 deletion mutants. Phosphorylation of full-length and some of the mutated pUL97 was detected in cells infected with the rVVs. The UL97 constructs carrying point mutations from GCV-resistant HCMV isolates at positions 460M, 520H, 594V, and the 4 aa deletion 590AACR593, also resulted in decreased but not abolished phosphorylation of GCV in the rVV system, whereas the phosphorylation of pUL97 itself was not influenced. The rVV system is a suitable method for quantitatively testing the functional relevance of pUL97 mutations. PMID- 9747719 TI - Transcription of the human cytomegalovirus natural killer decoy gene, UL18, in vitro and in vivo. AB - Multiplex RT-PCR analysis of human cytomegalovirus (HCMV) replication in human fibroblasts showed transcription of the natural killer (NK) cell decoy gene, UL18, from 72 h onwards. Transcription of glycoprotein B (gpUL55; a late gene) occurred from early time-points and peaked at 24 h post-infection. UL18 mRNA was also detected in the peripheral blood mononuclear cells of organ transplant recipients with HCMV viraemia, especially those with HCMV DNA virus loads greater than 10(5) genomes/ml whole blood. Thus, UL18 is produced via a low abundance transcript late during the infectious cycle at a time coincidental with the increased risk of NK cell lysis as a consequence of class I HLA down-regulation. PMID- 9747720 TI - NF-kappaB only partially mediates Epstein-Barr virus latent membrane protein 1 activation of B cells. AB - The latent membrane protein 1 (LMP1) of Epstein-Barr virus (EBV) is required for EBV-induced immortalization of human B cells and causes tumorigenic transformation of cell lines. LMP1 expression induces phenotypic changes resembling B cell activation, such as cell size increase and up-regulation of cell surface activation markers. LMP1 contains two domains that activate the transcription factor NF-kappaB, one through interactions with TRAF proteins and the other with the TRADD protein. The purpose of the present study was to investigate the importance of NF-kappaB induction in the up-regulation of the B cell activation markers ICAM-1 and CD71 by LMP1. This study shows that expression of LMP1 activates transcription from p50/p65- and c-Rel-responsive promoters, and that this activity can be completely inhibited by expression of a dominant inhibitory IkappaB mutant. ICAM-1 and CD71 are nevertheless up-regulated by LMP1 in primary B cells and cell lines expressing the dominant IkappaB. Furthermore, LMP1-induced cell size increase of primary B cells was unaffected by IkappaB expression. It was concluded that even when LMP1 is unable to activate NF-kappaB, it is still capable of inducing certain characteristics of activated B cells, strongly suggesting that LMP1 can also activate cells independently of NF-kappaB. PMID- 9747721 TI - Bovine papillomavirus transmission and chromosomal aberrations: an experimental model. AB - Enzootic haematuria and urinary bladder cancer in cattle are associated with feeding on bracken fern and bovine papillomavirus (BPV) infection. An increased rate of chromosomal aberrations in peripheral blood lymphocytes from chronically affected haematuric cows raised in bracken fern pastures has been reported, suggesting the presence of BPV in the peripheral blood of afflicted animals. The purpose of the present investigation was to examine the role of peripheral blood as a potential BPV-transmitting agent and search for clastogenic effects in experimentally infected animals kept on a bracken fern-free diet. Healthy cows were inoculated with blood samples of haematuric animals every two weeks for 18 months. Recipient cows, their offspring, donor animals and a control group were kept on a bracken fern-free diet throughout the experiment. Clinical and molecular analyses for detection of BPV infection were carried out periodically in all groups. Short-term lymphocyte cultures were performed to assess chromosomal aberration levels. The donor cows, the recipient cows and their offspring presented increased levels of chromosomal aberrations. BPV-2 DNA was identified by Southern blotting, PCR and cycle-sequencing of PCR products in peripheral blood of donor and recipient animals and in the progeny of recipient animals. Data support both the concept that BPV can be transmitted through blood and the hypothesis that infection with the virus causes the clastogenic alterations observed in the present experimental model. The presence of BPV-2 DNA and chromosomal alterations in peripheral blood of offspring at the moment of birth is evidence for vertical transmission of BPV. PMID- 9747723 TI - T-antigen-dependent transcriptional initiation and its role in the regulation of human neurotropic JC virus late gene expression. AB - The multifunctional protein of papovaviruses, T-antigen, regulates the virus lytic cycle partly by exerting transcriptional control over viral and cellular gene expression. In this study, the ability of the T-antigen of human neurotropic JC virus (JCV) to enhance expression from the virus late promoter has been further examined. By deletion analysis, a T-antigen-responsive region was mapped within the JCV 98 bp enhancer/promoter between nucleotides 139 and 168. Interestingly, T-antigen appears to mediate transactivation by increasing expression from a basal transcriptional initiation site and through a novel T antigen-dependent initiation site (TADI). The TADI element contains a region homologous to initiator (Inr) sequences and is sufficient to confer T-antigen responsiveness to a heterologous minimal promoter. Electrophoretic mobility shift and UV crosslinking analyses demonstrate that multiple cellular proteins interact with both single- and double-stranded forms of this sequence. Mutations within the TADI element which abolish T-antigen-mediated transcriptional activation also prevent the formation of specific nucleoprotein complexes. These data suggest that the ability of JCV T-antigen to regulate JCV late gene expression may be partly due to the formation of specific nucleoprotein complexes and transcriptional initiation from the TADI site on the viral promoter. PMID- 9747722 TI - Mutant canine oral papillomavirus L1 capsid proteins which form virus-like particles but lack native conformational epitopes. AB - Recently, the L1 capsid protein of canine oral papillomavirus (COPV) has been used as an effective systemic vaccine that prevents viral infections of the oral mucosa. The efficacy of this vaccine is critically dependent upon native L1 conformation and, when purified from Sf9 insect cells, the L1 protein not only displays type-specific, conformation-dependent epitopes but it also assembles spontaneously into virus-like particles (VLPs). To determine whether VLP formation was coupled to the expression of conformation-dependent epitopes, we generated a series of N- and C-terminal L1 deletion mutants and evaluated their ability to form VLPs (by electron microscopy) and to react with conformation dependent antibodies (by immunofluorescence microscopy). We found that (a) deletion of the 26 C-terminal residues generated a mutant protein which formed VLPs efficiently and folded correctly both in the cytoplasm and in the nucleus; (b) further truncation of the L1 C terminus (67 amino acids) resulted in a capsid protein which formed VLPs but which failed to express conformational epitopes; (c) deletion of the first 25 N-terminal amino acids also abolished expression of conformational epitopes (without altering VLP formation) but the native conformation of this deletion mutant could be restored by the addition of the human papillomavirus type 11 N terminus. These results demonstrate that VLP formation and conformational epitope expression can be dissociated and that the L1 N terminus has a critical role in protein folding. In addition, it appears that correct L1 protein folding is not dependent upon the nucleoplasmic environment. PMID- 9747724 TI - Transcription-positive cofactor 4 enhances rescue of adeno-associated virus genome from an infectious clone. AB - While Rep proteins are required for adeno-associated virus (AAV) replication, little is known about cellular proteins that interact with Rep. We demonstrate here that transcription-positive cofactor 4 (PC4, p15) fused to Gal4-activating domain interacted with both AAV-2 and AAV-3 Rep proteins fused to Gal4 DNA binding domain, leading to reporter activation in the yeast two-hybrid system. In addition to its coactivating function, PC4 recently has been shown to be involved in replication of simian virus 40. To study a functional role for the PC4-Rep protein interaction, 293-31 cells were cotransfected with a PC4 expression plasmid and an infectious clone of AAV-3, followed by super-infection with helper adenovirus. A significantly increased number of AAV-3 genomes were rescued in PC4 transfected cells. Our results support a possible involvement of PC4 in AAV replication and may be used in efficient production of AAV vectors for gene therapy. PMID- 9747725 TI - Binding of bovine parvovirus to erythrocyte membrane sialylglycoproteins. AB - Bovine parvovirus (BPV), an autonomous parvovirus, haemagglutinates human type O erythrocytes and infects certain bovine cells in culture. Little is known about the receptor to which it attaches, either on nucleated host cells or on erythrocytes. Haemagglutination assays and radiolabelled virus-binding tests measuring the effects of trypsin, chymotrypsin, neuraminidase, phospholipase C and sodium periodate on attachment of BPV to receptors indicated that BPV interacted with N-acetylneuraminic acid-containing (sialyl) glycoproteins. SDS polyacrylamide gel separation of erythrocyte ghost proteins and virus overlay protein-binding revealed BPV binding to glycophorin A. Confirmation testing showed BPV binding to purified glycophorin A on dot blots and on gels containing membrane glycophorin A and purified glycophorin A. Further, in competition assays, purified glycophorin A completely inhibited the BPV haemagglutination reaction. The results of this study indicate that BPV binds to sialated membrane glycoproteins, one of which is the major erythrocyte membrane glycoprotein, glycophorin A. PMID- 9747726 TI - Characterization of novel circovirus DNAs associated with wasting syndromes in pigs. AB - Porcine circovirus (PCV) was initially recognized as a contaminant of continuous pig kidney cell lines and was not thought to be pathogenic. Antibodies reactive to the cell culture isolate of PCV (PCV PK-15) are prevalent in the swine population worldwide. Recently, PCV PK-15-like antigen and nucleic acid were demonstrated in lesions associated with wasting syndromes in pigs in North America and Europe. Monoclonal antibodies raised to circoviruses isolated from pigs with wasting syndromes highlighted differences between these circoviruses and the PCV PK-15 cell culture isolate. This has led to speculation that a new pathogenic PCV may have emerged in the swine populations of several countries. We report the cloning and characterization of novel circovirus DNAs purified from virus isolates made from tissues of North American and European pigs with wasting syndromes. These North American and European circoviruses form a closely related group at the nucleotide sequence level (> 96% intra-group nucleotide sequence identity) but exhibit < 80% nucleotide sequence identity with the PCV PK-15 cell culture isolate. This report provides evidence for a new type of possibly pathogenic PCV. We propose that these new circoviruses should be referred to as PCV2 as opposed to the original PK-15 cell culture isolate, which should be referred to as PCV1. PMID- 9747727 TI - Translational regulation of hepatitis B virus polymerase gene by termination reinitiation of an upstream minicistron in a length-dependent manner. AB - Hepatitis B virus (HBV) polymerase (P) gene is translated from the bicistronic pregenomic RNA with the core (C) gene in the first cistron. The P ORF is preceded by the C AUG and three AUG codons within the C region, where a minicistron of 7 amino acids can potentially be translated. Our results indicate that the efficiency of the P gene translation initiation was about 10% of that of the C gene when both genes were fused in-frame to a lacZ reporter in an mRNA similar in structure to the pregenomic RNA. By mutational analysis, about 74% of the translation initiation of HBV P gene was shown to be by ribosomes that reinitiated after terminating translation of this minicistron, while the rest was by two mechanisms: one by ribosomes leaky scanning through every upstream AUG and the other by ribosomal backwards scanning to the P AUG after finishing the translation of the C gene. The efficiency of termination-reinitiation depended on the size of the minicistron, i.e. the reinitiation efficiency decreased about 50% when the size increased from 24 nt to 57 nt. When a 44 nt HBV sequence comprising the minicistron was inserted at the 5' untranslated region of the cat gene, CAT expression was regulated in a similar way to that of the HBV P gene. Moreover, when transfection occurred with an HBV expression plasmid containing an inactivated minicistron, production of virus-like particles dropped to about one third of the wild-type level, suggesting that the termination-reinitiation mechanism is indeed important for HBV P gene expression. PMID- 9747728 TI - Characterization of HLA-B57-restricted human immunodeficiency virus type 1 Gag- and RT-specific cytotoxic T lymphocyte responses. AB - HLA-B57 has been shown to be strongly associated with slow disease progression in human immunodeficiency virus type 1 (HIV-1)-infected patients from the Amsterdam Cohort. Since HIV-1-specific CTL can control and eliminate virus-infected cells, we sought to characterize the dominant HLA-B57-restricted CTL responses at the epitope level. It was found that HLA-B57-restricted CTL responses were targeted at multiple proteins of HIV-1, with CTL specific for Gag and RT being the most pronounced. Gag-specific CTL recognized peptides ISPRTLNAW (aa 147-155) and STLQEQIGW (aa 241-249), which had previously been reported as HLA-B57-restricted. The RT-specific CTL response in one long-term survivor studied in great detail persisted for > 10 years and was dominated by HLA-B57-restricted CTL that recognized the newly defined epitope IVLPEKDSW (RT(LAI), aa 244-252). This epitope could be recognized in the context of both HLA-B*5701 and HLA-B*5801. Interestingly, three epitope variants of IVLPEKDSW were observed, which coincided with the strongest detectable CTL response to RT. One variant (T2E7) was not recognized by IVLPEKDSW-specific CTL despite the fact that this variant bound to HLA-B*5701 with a similar affinity as the index peptide. Finally, only viruses which contained the epitope index sequence were obtained suggesting efficient virus control by CTL. In conclusion, we report the characterization of dominant HIV-1 Gag- and RT-derived, HLA-B57-restricted CTL epitopes which are associated with longer time to AIDS. Further characterization of CTL responses restricted by HLA-B57 and other protective HLA alleles may contribute to the development of effective AIDS vaccines. PMID- 9747729 TI - The simian immunodeficiency virus mnd(GB-1) strain uses CXCR4, not CCR5, as coreceptor for entry in human cells. AB - The simian immunodeficiency virus (SIV) mnd(GB-1) strain, isolated from a mandrill, replicates in a human T cell line, CEM cells, and is inhibited by the CXC-chemokines, stromal cell-derived factor 1alpha and 1beta (SDF-1alpha/SDF 1beta), the natural ligands for CXCR4. The IC50 was around 70-80 ng/ml, which corresponds to the IC50 of SDF-1alpha/SDF-1beta for T-tropic human immunodeficiency virus type 1 (HIV-1) and HIV-2. The specific anti-CXCR4 MAb 12G5 inhibited replication of SIVmnd at an IC50 of 1 microg/ml. Also, the IC50 of 8 ng/ml for SIVmnd of the bicyclam AMD3100, a specific CXCR4 antagonist, is comparable with its IC50 for T-tropic HIV-1 and HIV-2 strains. Two other SIV strains, SIVagm3 and SIVmac251, were insensitive to SDF-1alpha/SDF-1beta, anti CXCR4 MAb and AMD3100. SIVmnd replicates only in HOS.CD4 cells expressing CXCR4 and not in HOS.CD4 transfectants expressing CCR1, CCR2b, CCR3, CCR4 or CCR5. This is, to our knowledge, the first SIV strain found to use CXCR4 and not CCR5 as a main coreceptor for entering human cells. PMID- 9747730 TI - Identification of a 374 amino acid protein encoded by RNA segment 6 of influenza C virus. AB - Unspliced mRNA from RNA segment 6 of influenza C virus contains a single open reading frame that potentially encodes a polypeptide of 374 amino acids. This polypeptide, which has not been identified as yet, is predicted to contain the complete amino acid sequence of the matrix protein, M1, as well as that of a small integral membrane protein, CM2. Here, we found that small amounts of two previously unrecognized proteins with apparent molecular masses of 42 (P42) and 44 kDa (P44) were immunoprecipitated with immune serum against CM2. The electrophoretic mobilities of P42 and P44 varied depending on virus strain, indicating that they are virus-coded. Treatment of infected cells with tunicamycin and digestion of immunoprecipitated proteins with various endoglycosidases revealed that P42 is modified by the addition of a high-mannose oligosaccharide chain to generate P44. A monoclonal antibody against M1, like anti-CM2 serum, was able to immunoprecipitate both the P42 and P44 proteins. Furthermore, the tryptic peptide map of either P42 or P44 was indistinguishable from the map of the mixture of M1 and CM2. These results, taken together, suggest strongly that P42 and P44 correspond to the 374 amino acid protein encoded by unspliced RNA segment 6 mRNA and its N-glycosylated form, respectively. PMID- 9747731 TI - Enhanced neutralization of human respiratory syncytial virus by mixtures of monoclonal antibodies to the attachment (G) glycoprotein. AB - Neutralization of human respiratory syncytial virus (HRSV) by monoclonal antibodies specific for the G protein was evaluated in a microneutralization test. Only certain antibodies showed some degree of neutralization, reflected in a reduction of virus titre (10-20 times maximum), compared with negative controls. In contrast, a pool of antibodies that recognized conserved, group specific and strain-specific epitopes showed a significant increase in virus neutralization (up to 500-1000 times). By testing binary, tertiary and quaternary combinations, four antibodies were identified which showed maximal effect in the neutralization test. These findings are discussed in terms of the location of antibody binding sites in the G protein primary structure and their relevance for HRSV neutralization and immunobiology. PMID- 9747732 TI - Circulation patterns of genetically distinct group A and B strains of human respiratory syncytial virus in a community. AB - Human respiratory syncytial virus (HRSV) is classified into two major groups, A and B, each of which contains multiple variants. To characterize the molecular epidemiology of HRSV strains over time, sequencing studies of a variable region of the attachment protein gene from a single community in the United States during 5 successive years were performed. Phylogenetic analysis revealed distinct clades (genotypes) that were further classified in subtypes based on > or = 96% nucleotide similarity. Five genotypes and 22 subtypes among 123 group A HRSV isolates, and four distinct genotypes and six subtypes among 81 group B HRSV isolates were identified. One to two genotypes or subtypes accounted for > or = 50% of isolates from a given year. A shift in the predominant genotype or subtype occurred each year such that no genotype or subtype predominated for more than 1 of the 5 study years. The consistency in the displacement of the predominant strain suggests that a shift, even within the same group, is advantageous to the virus. It was hypothesized that the 'novel' strain is better able to evade previously induced immunity in the population and consequently either circulates more efficiently or is more pathogenic. The yearly shift in HRSV strains may contribute to the ability of HRSV to consistently cause yearly outbreaks of HRSV disease. These results also suggest that isolates may need to be characterized as to both group and genotype to fully understand protective immunity after natural infection and efficacy studies of candidate vaccines. PMID- 9747733 TI - Sequence analysis of a functional polymerase (L) gene of bovine respiratory syncytial virus: determination of minimal trans-acting requirements for RNA replication. AB - The complete nucleotide sequence of a functional clone of the large polymerase (L) gene of bovine respiratory syncytial virus (BRSV) strain A51908 was determined by analysis of cloned cDNAs obtained from genomic and mRNAs. The BRSV L gene is 6573 nt in length and the derived polypeptide has 2162 aa. Alignment of the sequences of the BRSV L gene, and its encoded protein, with sequences of the L gene and protein of human respiratory syncytial virus strain A2 showed 77% identity at the nucleotide level and 84% identity at the amino acid level. By comparison, the L gene and protein of avian pneumovirus showed only 50% identity at the nucleotide level and 64% identity at the amino acid level. A minigenome was constructed to encode a BRSV vRNA analogue containing the gene for chloramphenicol acetyltransferase (CAT) under the control of putative BRSV transcription motifs and flanked by the BRSV genomic termini. Transfection of plasmids encoding the BRSV minigenome, nucleocapsid protein (N), phosphoprotein (P) and L protein, each under the control of T7 promoter, into cells infected with a vaccinia virus recombinant expressing the T7 RNA polymerase gave rise to CAT activity and progeny with the minigenome. This result indicates that the N, P and L proteins are necessary and sufficient for transcription and replication of the BRSV minigenome and are functional. Further, inclusion of small amounts of the M2 protein along with the N, P and L proteins greatly augmented minigenome transcription. PMID- 9747734 TI - Constitutive overexpression of the major inducible 70 kDa heat shock protein mediates large plaque formation by measles virus. AB - Induction of the cellular stress response elevates cytoplasmic levels of heat shock proteins (HSPs) belonging to multiple families. When infected with canine distemper virus or measles virus (MV), cells containing elevated HSPs support increased viral gene expression and cytopathic effect. The present work tests the hypothesis that increases in the major inducible 70 kDa HSP (hsp72) are sufficient to mediate the effect of stress response induction on infection phenotype. Human astrocytoma cells (U373) were stably transfected with the human hsp72 gene under control of the beta-actin promoter. Constitutive overexpression of hsp72 was demonstrated in multiple clones by Western blot analysis of cytoplasmic total protein. Southern blot analysis of cell DNA confirmed the recovery of genetically distinct clones. Infection of these clonal populations with MV resulted in increased viral transcript production relative to infected control cell lines. Increased transcript production was associated with increased viral membrane glycoprotein expression and cytopathic effect (i.e., mean plaque area). Increases in cytopathic effect were due to the emergence of a large plaque phenotype from a small plaque-purified inoculum, mimicking the effect of cellular stress response induction upon viral infection phenotype. Large plaque phenotypic variants reported in the literature are associated with enhanced neurovirulence, a fact that highlights the potential significance of physiologic elevations in hsp72 (e.g., fever-induced) that accompany in vivo viral infection. PMID- 9747735 TI - Induction of neutralizing antibodies by synthetic peptides representing the C terminus of coxsackievirus A9 capsid protein VP1. AB - The arginine-glycine-aspartic acid motif at the C terminus of coxsackievirus A9 capsid protein VP1 has been shown to play a role in specific attachment of the virus to alpha(v)beta3 integrin on the host cell surface. The C-terminal region of the VP1 protein has also been shown to be highly antigenic by using peptide scanning techniques. To find out whether this region contains a neutralizing epitope, three overlapping peptides covering the C-terminal end of VP1 were synthesized and rabbit antisera were raised against these peptides. Neutralization of the virus was observed with all three antipeptide antisera in A549 cells and with two antisera in RD cells. PMID- 9747736 TI - Antiviral activity of bovine collectins against rotaviruses. AB - The antiviral activity against rotaviruses of three bovine collectins, conglutinin, collectin-43 (CL-43) and bovine SP-D, was examined. As shown by ELISA and Western blot, all three collectins bound to the Nebraska calf diarrhoea virus bovine strain of rotavirus, and specifically to the VP7 glycoprotein. Inhibition by mannose or EDTA confirmed that binding was mediated through the lectin domains of the collectins. Binding resulted in haemagglutination inhibition and neutralization of rotavirus infectivity, CL-43 displaying the highest activity in both types of assay. In contrast, conglutinin was the most potent of the three collectins against influenza virus A/HKx31. Neutralization of rotaviruses by the lectins was dependent on glycosylation of VP7. Furthermore, rotaviruses adapted to growth in Madin-Darby bovine kidney cells, and thus bearing carbohydrate of bovine origin, remained sensitive to neutralization, although slightly less so than virus stocks propagated in the monkey kidney cell line MA104. These findings provide the first description of antiviral activity of collectins against a non-enveloped virus and may indicate a potential role for collectins in host defence against bovine rotavirus infection. PMID- 9747737 TI - Mutational analysis of bean yellow dwarf virus, a geminivirus of the genus Mastrevirus that is adapted to dicotyledonous plants. AB - Bean yellow dwarf virus (BeYDV) is an atypical member of the geminivirus genus Mastrevirus that infects dicotyledonous plants. BeYDV DNA contains six open reading frames (ORFs) with the capacity to encode proteins in excess of 10 kDa. Two virion-sense ORFs (V1 and V2) and two complementary-sense ORFs (C1 and C2) have homologues in all mastreviruses, while ORFs C3 and C4 are not conserved. To investigate their functions, each of the ORFs has been truncated by either frame shifting or the introduction of a stop codon. We demonstrate that an ORF V1 mutant replicated efficiently in Nicotiana tabacum protoplasts but was unable to systemically infect Phaseolus vulgaris and Datura stramonium, consistent with a role for V1 protein in virus movement. However, the mutant was able to systemically infect Nicotiana benthamiana although the onset of symptoms was appreciably delayed in comparison with wild-type virus. Disruption of ORF V2, encoding the coat protein, prevented systemic infection of all three hosts but the mutant replicated in protoplasts. Both ORF C1 and ORF C2 were essential for replication in protoplasts. Modification of the complementary-sense splice donor and acceptor sequences also prevented replication. Removal of the intron prevented systemic infection, although the intronless mutant was able to produce functional replication-associated protein (Rep) and replicated efficiently in protoplasts. ORFs C3 and C4 were not required for systemic infection. Our results indicate that four ORFs are spatially and functionally conserved in mastreviruses that infect both monocotyledonous and dicotyledonous plants. PMID- 9747739 TI - Identification and characterization of the Cydia pomonella granulovirus cathepsin and chitinase genes. AB - A 3.2 kb BamHI-EcoRI fragment of the Cydia pomonella granulovirus (CpGV) genome was subcloned and characterized. Sequence analysis revealed two complete and one partial open reading frames (ORFs). ORF7L is predicted to encode a 66.7 kDa protein (594 amino acid residues) that is 57% identical (amino acid sequence) to the chiA gene (ORF126) of Autographa californica nucleopolyhedrovirus (AcMNPV), encoding a chitinase. ORF8R is 333 amino acids in length and shows high similarity (between 64% and 67%) with baculovirus cathepsins. The partial ORF, ORF5L, is related to AcMNPV ORF145 of unknown function. Phylogenetic trees were constructed for both chitinase and cathepsin sequences from baculoviruses and other species. In both cases, the baculovirus sequences were monophyletic but with a deep division between the GVs and NPVs, suggesting both genes were present in an ancestral virus prior to the separation of the two genera. However, these studies did not provide definitive evidence for the origin of either protein in baculoviruses. To investigate CpGV cathepsin function, a rescue experiment was performed using a Bombyx mori NPV (BmNPV) mutant (BmCysPD) which lacks a functional cathepsin (cath) gene. Larvae infected with BmCysPD-Cp.cat, a BmCysPD derivative carrying CpGV cath, showed similar symptoms to wild-type BmNPV infected insects, confirming that CpGV cath encodes a functional cathepsin. Primer extension analysis of mRNA from BmCysPD-Cp.cat infected cells showed that CpGV cath transcription was initiated from a consensus late transcription motif (ATAAG) within the CpGV sequences, indicating that a CpGV late promoter motif was recognized in this NPV system. PMID- 9747738 TI - cDNA cloning and molecular characterization of cherry green ring mottle virus. AB - The complete nucleotide sequence of the cherry green ring mottle virus (CGRMV) genome was determined to be 8372 nt excluding a 3' poly(A) tail. Based on computer analysis and sequence comparison, five open reading frames (ORFs) were identified on the virion strand encoding: a putative RNA-dependent RNA polymerase, a triple gene block and a coat protein. Two other ORFs with Mr values over 10,000 and internal to the helicase and coat protein genes, but of unknown function, were also identified. Sequence and genome structure comparisons with other filamentous viruses indicated that CGRMV is most similar to apple stem pitting virus, some carlaviruses and potexviruses. However, it is different from members of any of these virus groups in regard to sequence homology and genome organization. A chimeric fusion coat protein was expressed in E. coli and antibodies specific for the CGRMV coat protein were raised in rabbits. The antibody was used in Western blot analyses to detect the CGRMV coat protein in infected cherry tissue. PMID- 9747740 TI - Apoptosis resulting from superinfection of Heliothis zea virus 1 is inhibited by p35 and is not required for virus interference. AB - Superinfection of Spodoptera frugiperda insect cells that are persistently infected with Heliothis zea 1 (Hz-1) virus induces general cellular apoptosis and subsequently results in homologous virus interference. Since apoptosis correlates closely with both a significant decrease in yield of virus progeny and expansion of virus infection among cells, further experiments were designed to verify the direct association of apoptosis with homologous interference. It was found that superinfection-induced apoptosis can be efficiently blocked by the stable transfection of p35 into cells before or after the establishment of persistent virus infection. However, persistently infected cells are still strongly resistant to the challenge of Hz-1 virus, indicating that the induction of apoptosis is not essential for the resulting homologous Hz-1 virus interference. Replication and transcription of viral genomes are greatly retarded upon Hz-1 virus superinfection of persistently infected cells, whether stably transfected with p35 or not, suggesting that upon superinfection, the decreasing yield of virus progeny in these persistently infected cells is caused by a blockage early after virus infection. PMID- 9747741 TI - Survival in breast cancer patients after the first episode of hypercalcaemia. AB - OBJECTIVES: To investigate hypercalcaemia (serum ionized calcium (S-Ca2+) > 1.35 mmol L(-1)) in breast cancer patients before and after the introduction of bisphosphonates and the effect of disease- and treatment-related factors on survival. DESIGN: Prospective and retrospective registration of covariates. SETTING: A department of oncology in a university hospital. SUBJECTS: A consecutive cohort of 212 hypercalcaemic patients never treated with bisphosphonate was identified prospectively (period 1) and 193 patients with metastases were classified into three groups: mild (S-Ca2+ < 1.48: n=102). moderate (1.48 < or = S-Ca2+ < or = 1.60; n=41). and severe hypercalcaemia (S Ca2+ > 1.60 mmol L(-1); n=50). Fifty-one patients with severe hypercalcaemia all treated with bisphosphonate except one were identified retrospectively (period 2). RESULTS: For period 1 median survival was 6.7 months. Survival was significantly decreased in the two groups with the highest initial S-Ca2+ (P < 0.0001). Median survival times in severely hypercalcaemic patients from periods 1 and 2 were 1.4 (9 5(% confidence interval 0.8-2.1) and 2.2 (95% confidence interval 1.3-3.1) months, respectively. In a Cox model for period 1 significant covariates were: WHO performance, extent of metastases, whether systemic anticancer treatment could be given, and haemoglobin, but not S-Ca2+. CONCLUSION: Prognosis is poor in hypercalcaemic breast cancer patients with WHO performance 3 4 and advanced metastatic disease when effective systemic treatment can no longer be offered. Bisphosphonate treatment does not seem to improve survival in severe hypercalcaemia. Antihypercalcaemic treatment of mild malignancy-associated hypercalcaemia appears not to be vital. Therapeutic efforts should be aiming at patients with moderate hypercalcaemia. PMID- 9747742 TI - Diagnosis of pulmonary embolism by measurement of alveolar dead space. AB - OBJECTIVE: Pulmonary embolism (PE) gives rise to alveolar dead space, which can be measured with a single breath test for CO2 (SBT-CO2). The characteristics of the SBT-CO2 are different in PE and other common conditions giving rise to alveolar dead space, notably airways disease. An analysis of alveolar dead space focusing on the late part of the breath (fDlate) has been suggested as a method for diagnosis of PE. Our aim was to evaluate this technique by comparison with lung scintigraphy. METHODS: We randomly selected patients with clinical suspicion of PE. SBT-CO2 and lung scintigraphy were performed on the same day. The scintigraphies were reviewed and classified as high, intermediate and low probability of PE. RESULTS: Out of 223 patients able to be evaluated, there were 20 of the high, 29 of the intermediate and 174 of the low probability category. There were large differences between the means of fDlate in the high and the intermediate and in the high and the low categories. We obtained a sensitivity of 85% and a specificity of 93% for diagnosis of PE, based on high and low probability categories. If a patient with previous PE, but no scintigraphic evidence of current PE, is excluded the sensitivity increases to 90%. CONCLUSIONS: This study provides further support for the measurement of fDlate by the SBT-CO2 as a diagnostic test in patients with suspicion of PE. The test should be especially useful in small hospitals without access to pulmonary scintigraphy or pulmonary angiography. PMID- 9747743 TI - Disease spectrum of patients with antineutrophil cytoplasmic autoantibodies of defined specificity: distinct differences between patients with anti-proteinase 3 and anti-myeloperoxidase autoantibodies. AB - OBJECTIVE: To compare the disease spectrum of consecutive patients with antineutrophil cytoplasmic autoantibodies directed against proteinase 3 (anti PR3) or myeloperoxidase (anti-MPO). DESIGN: Retrospective analysis. SETTING: Three teaching hospitals in the Netherlands. MAIN OUTCOME MEASURES: Clinical features at presentation, histopathological characteristics and outcome. SUBJECTS: All consecutive patients who tested positive for anti-PR3 (n=46) or anti-MPO (n=46) over an 8-year-period. RESULTS: At diagnosis, patients with anti PR3 had a higher vasculitis activity index than patients with anti-MPO (P < 0.001). The mean (SD) number of affected organs in the anti-PR3 group exceeded that of the anti-MPO group (3.9 (1.4) and 2.2 (1.1), respectively; P < 0.01). The combination of renal and respiratory tract involvement was present in as many as 78.3% of patients with anti-PR3 and in only 23.9% of patients with anti-MPO (P < 0.01). Renal-limited disease exclusively occurred in patients with anti-MPO. Granulomas were found in 41.3% of anti-PR3- but in only 4.3% of anti-MPO-positive patients (P < 0.01). All anti-PR3-positive patients had Wegener's granulomatosis or microscopic polyangiitis. By contrast, diagnoses in the anti-MPO group were more diverse: idiopathic necrotizing crescentic glomerulonephritis (26.1%), microscopic polyangiitis (26.1%). Churg-Strauss syndrome (4.3%), Wegener's granulomatosis (2.2%), giant cell arteritis (2.2%), clinically suspected vasculitis (19.6%), as well as miscellaneous nonvasculitic disorders (19.6%). During follow-up, 10 anti-PR3-positive patients had 11 relapses whereas only 3 patients with anti-MPO relapsed (P=0.04). CONCLUSION: A large divergence was seen in the disease spectrum between patients with anti-PR3 and those with anti-MPO. In particular, extra-renal disease manifestations, granuloma formation and relapses were more prominent in anti-PR3- than in anti-MPO-positive patients. PMID- 9747744 TI - Prophylactic splenectomy and cholecystectomy in mild hereditary spherocytosis: analyzing the decision in different clinical scenarios. AB - OBJECTIVES: Patients with mild hereditary spherocytosis (HS), i.e. with haemolysis without anaemia. have an increased risk of gallstone formation, erythroid aplasia and haemolytic crisis. Since the effect of prophylactic splenectomy on life expectancy has not been established, we conducted a decision analysis comparing prophylactic splenectomy and cholecystectomy with no surgery. DESIGN: The available data on surgery and disease outcomes were modelled to estimate the effects of the different interventions on the quality-adjusted life expectancy. The early phase outcomes depicted surgery-related mortality and incorporated compliance and the adverse effects of prophylaxis against post splenectomy infections. The late phase outcomes were framed by a Markov cohort analysis. RESULTS: For patients without gallstones, surgery was of no benefit. For those with gallstones the preferred strategies were found to be splenectomy and cholecystectomy before the age of 39 when asymptomatic, and before 52 when accompanied by occasional biliary colic. Cholecystectomy alone proved to be the preferred strategy in older patients with occasional biliary colic. For patients of up to 52 years of age and candidates for cholecystectomy because of recurrent biliary colic, the best strategy was to combine this procedure with splenectomy. Sensitivity analysis showed that the results were sensitive to the incidence of post cholecystectomy syndrome. Most remarkably an extreme sensitivity to compliance with post splenectomy infection prophylaxis was demonstrated. CONCLUSIONS: Our model suggested that combined prophylactic splenectomy and cholecystectomy provide a substantial gain in quality-adjusted life expectancy for young patients and adults with mild HS and gallstones. PMID- 9747745 TI - Humoral immune response to Chlamydia pneumoniae in twin discordant for smoking. AB - OBJECTIVES: The aim of this study was to investigate whether there is a relationship between smoking and Chlamydia pneumoniae specific antibody levels in generally healthy subjects, and whether this possible relationship is dose dependent. DESIGN: Match pair study. SUBJECTS: The study population comprised of 111 same-gender twin pairs from the Finnish Twin Cohort who in a previous study reported the highest discordance between smoking assessed as pack-years. MAIN OUTCOME MEASURES: Smoking and background data were obtained by a questionnaire, and C. pneumoniae specific serum IgG and IgA antibodies were measured by the micro-immunofluorescence (mIF) test. RESULTS: A significantly higher proportion of men with a history of smoking had elevated levels (a titre of > or=40) of serum IgA antibodies (P=0.003), whereas in women, a significant difference between the pairs was found in the proportion of IgG seropositive (a titre of > or=128) subjects (P=0.03). Conditional logistic regression analysis revealed that the risk for elevated IgA antibodies suggestive of chronic infection was significantly increased in current or former smokers in men (odds ratio 5.0 with 95% confidence intervals of 1.45-17.3). No dose-response effect was found between smoking and IgG or IgA titres, neither even if men and women were analysed separately. CONCLUSION: Smoking was significantly associated with elevated IgA antibody levels in men, supporting indirectly the hypothesis that smoking is a contributory factor in the establishment of chronic C. pneumoniae infection. PMID- 9747746 TI - Comparison between subcutaneous and intravenous interleukin-2 treatment in HIV disease. AB - OBJECTIVES: Therapy of HIV-infection using intravenous interleukin-2 (IL-2) is known to be effective in terms of increasing CD4-counts but is associated with significant side-effects and hospitalization. However, the combination with protease-inhibitor therapy has not been tested yet. The aim of the present study was to investigate the safety and efficacy of intravenous vs. subcutaneous IL-2 regimes using 9 Mio I.U. of IL-2, in combination with protease-inhibitor therapy. DESIGN: All patients were treated with a combination of two reverse transcriptase inhibitors and a protease-inhibitor prior to IL-2 administration for at least 6 weeks. Ten patients were assigned to the intravenous IL-2 group (group A). 10 to the subcutaneous group (group B). RESULTS: In both treatment groups, CD4 count significantly increased shortly after the end of therapy (group A: 223% over baseline [day 7]; group B: 264% over baseline [day 7]). During the follow-up CD4 counts slowly decreased thereafter but remained above baseline 3 months following IL-2 treatment. The CD8 lymphocytes showed a similar but less pronounced pattern with a maximum at day 7 (group A: 116% over baseline, group B: 158% over baseline) and reached baseline earlier in the follow-up-period. Altogether the CD4/CD8-ratio was elevated through long periods on follow-up. Throughout follow up, there were no apparent changes in viral load during IL-2 therapy in either groups. IL-2 therapy was administered for a mean time of 4.2+/-0.1 days in the intravenous group and of 4.8+/-0.1 days in the subcutaneous group until therapy was terminated at day 5 or due to side-effects. Only 1/10 patients completed the 5-day course of intravenous therapy in contrast to 6/10 in the subcutaneous group. CONCLUSIONS: Subcutaneous interleukin-2 using 9 Mio IU day(-1) in combination with protease-inhibitors showed equal efficacy as intravenous therapy and was associated with less side-effects. PMID- 9747747 TI - Metabolic abnormalities related to cardiovascular risk in primary hyperparathyroidism: effects of surgical treatment. AB - OBJECTIVES: Untreated primary hyperarathyroidism (pHPT) is accompanied by an excessive morbidity in circulatory disorders, associated with blood pressure and diabetes. The aim of the present study was to further penetrate the impact of pHPT on glucose, urate, lipid and lipoprotein concentrations, known to be interrelated metabolic cardiovascular risk factors. DESIGN: Longitudinal study of patients with pHPT before and 1 year after surgical treatment. SETTING: Departments of Internal Medicine and Surgery, Lund University Hospital. SUBJECTS: One hundred and seventeen consecutive patients with pHPT referred to surgical treatment. At presentation. 11 patients had previously diagnosed diabetes mellitus. INTERVENTION: All patients were successfully operated for pHPT. MAIN OUTCOME MEASURES: Fasting blood glucose and serum concentrations of cholesterol, triglyceride and urate were determined before and 1 year after surgery. The concentration of LDL- and HDL-cholesterol was separately analyzed in 21 cases. These data as well as the systolic and diastolic blood pressure were related to intact PTH and ionized calcium at presentation. Glomerular filtration was separately measured pre-operatively and related to the urate values. RESULTS: While the mean value for glucose remained unchanged among 11 patients with previously diagnosed diabetes at presentation, a significant decrease of glucose from 5.03+/-0.13 to 4.71+/-0.08 mmol/L (P < 0.05) was found among patients without known diabetes. Out of these patients, eight had diabetic glucose values at presentation, decreasing from 8.35+/-0.54 to 5.10+/-0.35 mmol/L (P < 0.05), and 12 had glucose values indicating impaired glucose tolerance, decreasing from 5.94+/-0.06 to 5.10 +/-0.38 mmol/L (P < 0.05) after surgery. Total cholesterol and trigylceride concentrations were not changed. However, male patients had significantly lower triglyceride levels at follow-up, 1.16+/-0.09 mmol/L compared to 1.57+/-0.14 mmol/L before surgery (P < 0.05). Significantly lower triglyceride values were also found among patients with glucose values indicating impaired glucose tolerance at presentation. The LDL/HDL cholesterol ratio remained normal. The serum level of urate decreased in both male and female patients after surgery, and was positively correlated to the PTH and ionized calcium values and inversely correlated to renal function before treatment. There was no significant correlation between calcium or PTH and the other metabolic variables studied. PMID- 9747748 TI - A nurse-managed weight reduction programme for obstructive sleep apnoea syndrome. AB - OBJECTIVES: This longitudinal, clinical intervention study was designed to investigate whether pulmonary departments can set up a cost-effective weight and lifestyle programme as primary treatment of obstructive sleep apnoea syndrome (OSAS). SETTING: A weight reduction programme (1 year) in a pulmonary department for outpatients in Helsinki University Central Hospital. SUBJECTS: A total of 24 (23 men) moderately obese (body mass index [BMI], 30-40 kg m(-2)) patients with newly diagnosed OSAS. Interventions. The first 6 weeks consisted of a very low calorie diet (VLCD, 500 kcal day(-1)) and thereafter normal food low in calories. There were altogether 12 group meetings for behavioural management. MAIN OUTCOME MEASURES: Daytime somnolence, BMI and oxygen desaturation index of 4% (ODI4) were measured prior to the programme, at the end of the VLCD phase and at 1 year. RESULTS: The programme was easy to administer without any serious side-effects. At 1 year, patients had lost a mean of 33% of their overweight (mean weight at baseline 110 kg, after 99 kg) and their ODI4 indexes improved significantly (P < 0.005). There was no correlation between the amount of weight loss and improvement in ODI4 indexes. The cost per patient was about half the cost of treatment with nCPAP (nasal continuous positive airway pressure) for 1 year at our hospital. CONCLUSIONS: A nurse-managed programme with VLCD and behavioural management is safe and effective on an outpatient basis. Weight loss should be encouraged in OSAS in patients with moderate overweight. The amount of weight loss needed for improvement of OSAS is unique to each individual. PMID- 9747749 TI - Acute lung injury due to parvovirus pneumonia. AB - OBJECTIVE: Human parvovirus B19 is responsible for the common childhood exanthematous illness, erythema infectiosum. Adults infected with B19 have been reported to develop a febrile illness associated with arthritis. Life threatening parvovirus infections in non-immunocompromised adults have not been reported to date. We report a previously healthy middle aged female who developed severe parvovirus pneumonia. The patient recovered with supportive care provided in the ICU. Parvovirus may represent an under diagnosed cause of community acquired pneumonia. PMID- 9747750 TI - Sheehan's syndrome: differential diagnosis in the acute phase. AB - Many studies have been done in the later course of Sheehan's syndrome, but very few have documented the acute phase with clinical, endocrine and imaging data. We present the case of a young woman complaining of severe headache after delivery, who developed hypopituitarism. Magnetic resonance imaging (MRI) disclosed the presence of an enlarged non haemorrhagic pituitary gland. Follow-up MRI showed a spontaneous and rapid shrinkage of the pituitary, within 20 days, which appeared as an empty sella 3 months later. Sheehan's syndrome may initially closely mimic hypophysitis, or the necrosis of an adenoma. We discuss the differential diagnoses, important for the best therapeutic management. PMID- 9747751 TI - Microbiological and epidemiological investigation of cholera epidemic in Ukraine during 1994 and 1995. AB - The Ukraine cholera epidemic of 1994 and 1995 was caused by Vibrio cholerae O1, serotype Ogawa, biotype El Tor. This epidemic was centred in the area around Respublika Krim (Crimea) and Mykolajiv, and spread to include parts of southern Ukraine. Cases of cholera occurred between September and November of 1994 and between June and October of 1995. The 32 fatalities among 1370 recorded cases (case fatality ratio, 2.3%) occurred throughout the course of the epidemic. V. cholerae from patients with cholera produced cholera toxin and were resistant to multiple antibiotics, though no resistance plasmids were found. Conjugation experiments suggested that resistance to multiple antibiotics may be present on a self-transmissible genetic element. Environmental sources of V. cholerae O1 El Tor included sewage, sea and surface water, and fresh water and marine fish. All but one of the environmental V. cholerae isolated during the epidemic were very similar to selected isolates from patients at the same time, supporting the role of these environmental sources in the spread of disease. PMID- 9747752 TI - Investigation of the 1994-5 Ukrainian Vibrio cholerae epidemic using molecular methods. AB - Thirty-seven Vibrio cholerae and four non-cholera Vibrio isolates from Ukraine, including strains from the epidemic of 1994-5, were analysed by molecular methods. Results from PFGE and ribotyping indicated that all Ukrainian toxigenic V. cholerae were closely related to each other and to an isolate from a patient from Pakistan. A non-toxigenic river water strain obtained during the height of the epidemic was more distantly related to these V. cholerae strains, while the Vibrio parahaemolyticus isolates and Vibrio alginolyticus isolate were not closely related to V. cholerae or each other. ERIC- and REP-PCR allowed the differentiation of strains identical by other methods. The results obtained confirm that the epidemic Ukrainian strains are most closely related to seventh pandemic strains from Asia and support a hypothesis that the Ukrainian epidemic of 1994-5 was caused by toxigenic environmental strains surviving since the time of the 1991 Ukrainian epidemic or before. PMID- 9747753 TI - Epidemiology and properties of heat-stable enterotoxin-producing Escherichia coli serotype O169:H41. AB - Enterotoxigenic Escherichia coli (ETEC) serotype O169:H41 organisms have become the most prevalent ETEC in Japan since the first outbreak in 1991. It was assumed that the outbreaks were due to clonal spread of this new ETEC serotype. The relationship of 32 strains isolated from 6 outbreaks were examined for biotype, antibiotic susceptibility, enterotoxigenicity, protein banding pattern, lipopolysaccharide banding pattern, plasmid analysis, and ribotyping. Further, the strains were examined by haemagglutination, surface hydrophobicity, and the ability to adhere to HEp-2 cells. The present study suggests that the outbreaks were caused by multiple clones of STp-producing O169:H41 since they showed differences in ribotype and outer membrane protein banding patterns. The strains did not agglutinate human or bovine red blood cells in a mannose-resistant manner. They adhered to HEp-2 cells in a manner resembling enteroaggregative E. coli. Five strains were examined by dot-blot tests for the colonization factor antigens CFA/I, CS1, CS2, CS3, CS4, CS5, CS6, CS7, PCFO159, PCFO166 and CFA/III. Although four strains expressed CS6, no structure for CS6 was identified. A strain that the anti-CS6 MAbs did not react with could adhere to HEp-2 cells in mannose resistant manner; thus, it is unlikely that CS6 play an important role in the adhesion to the cells. Electron microscopy studies of the O169:H41 strains suggested that curly fimbriae, a possible new colonization factor, may be playing an important role in the adhesion of the bacteria to HEp-2 cells. In conclusion, outbreaks due to ETEC O169:H41 were caused by multiple clones, and the strains should be examined in detail for a possible new colonization factor. PMID- 9747754 TI - Shigella infections among children in Andaman--an archipelago of tropical islands in Bay of Bengal. AB - Shigellosis is common among children in the Andaman and Nicobar islands. Our experience showed two distinct features of shigellosis within a span of 3 years in 1994-6: (i) changing patterns of serotype or subtype specific shigellosis and (ii) emergence of multidrug resistant isolates with changing R-patterns. The rate of isolation was 10.4-27.9% with the rate of isolation of Shigella flexneri interchanging with S. dysenteriae alternately. In 1994, S. flexneri superseded S. dysenteriae (48.6% vs. 33.3%; P < 0.05) while S. dysenteriae dominated over S. flexneri in 1995 (54.7% vs. 34.0%; P < 0.05). The picture reversed again in 1996 (63.0% vs. 22.2%; P < 0.05). Among shigellae isolates, the commonest serotypes were S. dysenteriae type 1 and S. flexneri type 2a. Isolated shigellae were of multidrug resistant type. Seven R-patterns were observed in 1994, while 8R patterns were observed during the next year with the emergence of nalidixic acid resistance. In 1996, emergence of gentamicin resistance was also observed. All isolates were resistant to ampicillin and sensitive to quinolones. The MIC of nalidixic acid and gentamicin are > or = 128 microg/ml and > or = 64 microg/ml respectively. These changing trends in shigellosis has important public health significance. PMID- 9747755 TI - Laboratory investigation and comparison of Salmonella Brandenburg cases in New Zealand. AB - An apparent increase in the incidence of S. Brandenburg in New Zealand, coupled with the possibility that the virulence of the organism may also be changing, prompted this study. Three typing methods: macro-restriction fragment length polymorphism (MRFLP) profiling using pulsed field gel electrophoresis (PFGE), plasmid profiling and antimicrobial susceptibility profiling were used to determine strain diversity amongst 115 recent and historical isolates of S. Brandenburg from both human cases and non-human sources. Antimicrobial resistance was noted only in three isolates. Plasmids of varying sizes were found in 31 isolates. MRFLP analysis resulted in 13 different patterns. Combining the three sets of typing data yielded 24 composite types. Comparison of composite type, isolation date and geographical location of case allowed the retrospective recognition of seven potential clusters during the 5-year study period. Composite types of 24 (80%) of the non-human isolates tested were indistinguishable from human isolates, suggesting that human infection may be via a number of vehicles. Although not cost-effective for routine use on all salmonella isolates, the methods used in this study are an important adjunct to serotyping for discrimination within an emerging serotype. PMID- 9747756 TI - Reduction of campylobacter infections in broiler flocks by application of hygiene measures. AB - Transmission routes of Campylobacter spp. in broilers and possibilities for prevention of infections were studied on two Dutch broiler farms. The occurrence of Campylobacter spp. was studied in successive broiler flocks, in the environment of the farms and in some of the parent flocks involved. Isolates of Campylobacter spp. were typed by using randomly amplified polymorphic DNA (RAPD) analysis. The results indicate that broiler flocks become infected from environmental sources. The typing results suggest that on one farm transmission of Campylobacter spp. occurred from cattle to broilers via the farmer's footwear. After several campylobacter positive broiler cycles hygiene measures, including thorough cleaning and disinfection procedures, change of footwear at the entrance of each broiler house, control of vermin and other hygienic precautions, were introduced on both farms in order to prevent transmission of Campylobacter spp. from the farm environment to the broilers. The results indicate that the application of hygiene measures significantly reduced campylobacter infections of broiler flocks on both farms. PMID- 9747757 TI - The prevalence of Vibrio spp. in drinking water and environmental samples in Vellore South India. AB - The prevalence of Vibrio cholerae in drinking water, lakes and sewage outfalls during July and August 1996 in Vellore, India was determined. Drinking water samples were collected on single occasions from 12 sites in different geographic areas of the town where cholera had been reported. Samples of water, plankton and sediment were collected from fixed sites at three lakes on three occasions separated by at least 3 days during the course of the study. Samples from open sewers were taken from two representative sites in four areas of the town. Bacteria isolated from samples were identified by standard biochemical tests and isolated strains of V. cholerae tested for their ability to agglutinate O1 and O139 antisera. Water samples from lakes were also tested for the presence of V. cholerae O1 and O139 by fluorescent antibody staining. Non-O1, non-O139 strains of V. cholerae were detected in 41% of drinking water samples and 100% of water, sediment and plankton samples from the test lakes. Eighty-seven per cent of open sewers sampled contained viable non-O1, non-O139 V. cholerae. Fluorescent antibody staining gave positive results for V. cholerae O1 and O139 for all water samples from the three lake sites. Strains of Aeromonas spp. were isolated from 58% of drinking water samples and from 66% of sediment, 77% of plankton and 55% of water samples from lakes. All open sewers sampled contained Aeromonas spp. PCR amplification employing specific primers demonstrated that none of the non agglutinating V. cholerae isolates contained the ctx operon. The non-O1, non-O139 V. cholerae isolates showed different patterns of antibiotic resistance to ampicillin, ciprofloxacin, chloramphenicol, tetracycline and trimethoprim. PMID- 9747758 TI - M genotyping and DNA fingerprinting of Streptococcus pyogenes isolates from an area of central Italy. AB - M protein gene typing was used to analyse Streptococcus pyogenes clinical isolates collected between 1983 and 1995 in an area of central Italy from patients presenting different types of infections; the same isolates were also characterized by means of DNA fingerprinting. M type 1 was the most common (50% of study strains), followed by M types 4, 12 and 6. The proportion of M type 12 decreased with time, whereas M type 1 increased, in agreement with data obtained in many different areas. Most invasive strains belonged to types M1 (30%) and M12 (30%); on the other hand, the M1 type did frequently occur also among non invasive isolates. DNA fingerprinting showed a correlation between M types and DNA patterns. This report provides epidemiological information from a geographic area not sampled recently, and further shows the usefulness of the M genotyping technique, which offers potential advantages over conventional serological typing methods. PMID- 9747759 TI - Dynamics of the meningococcal carrier state and characteristics of the carrier strains: a longitudinal study within three cohorts of military recruits. AB - Three cohorts of Danish male military recruits (n = 1069) were studied for pharyngeal meningococcal carriage during 3 months at different seasons: 39-47% of entrants were meningococcal carriers and the carriage rate remained constant over time and season. However, individual changes in the carrier state occurred frequently, and after 3 months 34% had changed carrier state on one or more occasions. Initially, a loss of carriage predominated; on the other hand almost 20% of non-carriers had acquisition of meningococci within the first month. The serological phenotypes of the 670 carrier strains were compared with those of 261 invasive strains recovered concurrently from patients with meningococcal disease country-wide. Both carrier strains and invasive strains were phenotypically heterogeneous. Almost 60% of the invasive strains belonged to three phenotypes: B:15:P1.7, 16, C:2a:P1.2, 5 and C:2b:P1.2, 5. In contrast, these phenotypes only amounted to 3.2% of the carrier strains, among which no phenotype was found with a prevalence above 4.9%. However, 30% of the carrier strains had serological phenotypes identical to those of 80% of the invasive strains. Our results indicated that the transmission rate of potential pathogenic carrier strains did not differ from that of other carrier strains. PMID- 9747760 TI - Molecular variation of meningococcal serotype 4 antigen genes. AB - Changes in the frequency of serogroup B non serotypable (B:NT) meningococci isolated in England and Wales were investigated by T-track fingerprint analysis, DNA nucleotide sequence determination, and serotyping by whole cell ELISA and dot blot assay. Seventy-three per cent of the isolates designated as B:NT by the Meningococcal Reference Unit (MRU) dot blot assay during 1993-4, expressed variants of the serotyping antigen, PorB, that were serotype 4 by whole cell ELISA. T-track fingerprint patterns of these and other 'serotype 4' isolates revealed five distinct porB alleles which were shown by nucleotide sequence determination to encode different peptide sequences. Differential binding of the 'serotype 4' mAbs MN14G21 and 5DC4C8G8 in whole cell ELISA and dot blot assays was the result, (i) of differences in the peptide sequence of predicted surface loop I and (ii) an amino acid deletion in predicted loop VI of the PorB protein. PMID- 9747761 TI - Transmission risk of Borrelia burgdorferi sensu lato from Ixodes ricinus ticks to humans in southwest Germany. AB - The risk of Borrelia burgdorferi infection and the value of antibiotic prophylaxis after tick bite are controversial. In this study, performed in two areas of southwestern Germany, ticks were collected from 730 patients and examined by the polymerase chain reaction (PCR) for B. burgdorferi. To assess whether transmission of B. burgdorferi occurred, the patients were clinically and serologically examined after tick removal and during follow-up examinations. Data from all tick bites gave a total transmission rate of 2.6% (19 patients). Eighty four ticks (11.3%) were PCR positive. Transmission occurred to 16 (26.7%) of 60 patients who were initially seronegative and could be followed up after the bite of an infected tick. These results indicate that the transmission rate from infected ticks in Europe is higher than previously assumed. Examination of ticks and antibiotic prophylaxis in the case of positivity appears to be indicated. PMID- 9747762 TI - Outbreaks of waterborne infectious intestinal disease in England and Wales, 1992 5. AB - Following the introduction of an improved surveillance system for infectious intestinal disease outbreaks in England and Wales, the Public Health Laboratory Service Communicable Disease Surveillance Centre received reports of 26 outbreaks between 1 January 1992 and 31 December 1995 in which there was evidence for waterborne transmission of infection. In these 26 outbreaks, 1756 laboratory confirmed cases were identified of whom 69 (4%) were admitted to hospital. In 19 outbreaks, illness was associated with the consumption of drinking water from public supplies (10 outbreaks) or private supplies (9 outbreaks). The largest outbreak consisted of 575 cases. In 4 of the remaining 7 outbreaks, illness was associated with exposure to swimming pool water. Cryptosporidium was identified as the probable causative organism in all 14 outbreaks associated with public water supplies and swimming pools. Campylobacter was responsible for most outbreaks associated with private water supplies. This review confirms a continuing risk of cryptosporidiosis from chlorinated water supplies in England and Wales, and reinforces governmental advice to water utilities that water treatment processes should be rigorously applied to ensure effective particle removal. High standards of surveillance are important for prompt recognition of outbreaks and institution of control measures. As microbiological evidence of water contamination may be absent or insufficient to implicate a particular water supply, a high standard of epidemiological investigation is recommended in all outbreaks of suspected waterborne disease. PMID- 9747763 TI - Seasonal trends of viral respiratory tract infections in the tropics. AB - To evaluate the seasonal trends of viral respiratory tract infections in a tropical environment, a retrospective survey of laboratory virus isolation, serology and immunofluorescence microscopy in two large general hospitals in Singapore between September 1990 and September 1994 was carried out. Respiratory tract viral outbreaks, particularly among infants who required hospitalization, were found to be associated mainly with respiratory syncytial (RSV) infections (72%), influenza (11%) and parainfluenza viruses (11%). Consistent seasonal variations in viral infections were observed only with RSV (March-August) and influenza A virus (peaks in June, December-January). The RSV trends were associated with higher environmental temperature, lower relative humidity and higher maximal day-to-day temperature variation. Although the influenza A outbreaks were not associated with meteorological factors, influenza B isolates were positively associated with rainfall. These data support the existence of seasonal trends of viral respiratory tract infections in the tropics. PMID- 9747764 TI - Estimating influenza-related hospitalization in The Netherlands. AB - The purpose of this study was to examine the impact of influenza on hospitalization in The Netherlands. Two methods were applied to estimate this effect: (a) regression analysis and (b) comparison of hospitalization in epidemic years with non-epidemic years. Hospital discharge rates in 1984-93 have been considered. The study shows that, during the period studied, on average, almost 2700 people were hospitalized for influenza per annum, and that influenza was diagnosed as the main cause for hospitalization in only a fraction of these hospitalizations (326: 12%). From an economic perspective, these results imply that the cost-effectiveness of vaccination against influenza may be severely underestimated when looking only at changes achieved in the number of hospitalizations attributed to influenza. PMID- 9747765 TI - Surveillance of small round structured virus (SRSV) infection in England and Wales, 1990-5. AB - Data from the national surveillance scheme for general outbreaks of intestinal disease, and the national laboratory reporting scheme were used to describe the epidemiology of small round structured virus (SRSV) infections in England and Wales. Between 1990 and 1995, there were 7492 laboratory reports of SRSV. Rates of reported illness were highest among infants, young children and the elderly. During 1992-5, some 707 SRSV outbreaks were reported. Outbreaks in hospital wards and residential facilities for the elderly accounted for 76% of the total, and annual numbers increased more than sixfold over the study period. There were wide regional variations in the numbers of SRSV outbreaks and laboratory reports. Both sporadic cases and outbreaks in the community are likely to be underestimated, but these passive surveillance systems provide an insight into the burden of SRSV infection among the institutionalized elderly. PMID- 9747766 TI - The 1993 dengue 2 epidemic in Charters Towers, North Queensland: clinical features and public health impact. AB - In 1993 an epidemic caused by dengue virus type 2 occurred in several North Queensland population centres. Charters Towers, estimated population 10,000, had 155 officially notified cases. An analysis of symptoms was undertaken using a random sample of 1000 residents to determine specificity of symptoms, the subclinical infection rate, and to establish the true extent of the epidemic. Retrospective diagnoses of dengue fever were based on the presence of both serum dengue 2 neutralizing antibody and presence of symptoms. An estimated 20% of the population had dengue fever. The rate of subclinical infections in this epidemic was 14.6%. There were no symptoms that were specific for dengue fever. Bleeding occurred more frequently in people who recalled a previous dengue infection during a dengue 1 epidemic 12 years earlier (55.6% vs. 16.8%, P = 0.003). Surveillance for future epidemics should be based on serological and virological confirmation of dengue virus infection amongst symptomatic patient. PMID- 9747767 TI - Implementation and evaluation of a measles/rubella vaccination campaign in a campus university in the UK following an outbreak of rubella. AB - An age shift in rubella infection to young adults has occurred in Scotland since the introduction of a first dose measles, mumps and rubella (MMR) vaccination in 1988 and a second dose measles/rubella (MR) vaccination in 1994/95. The Health Board was alerted to an outbreak of rubella at Stirling University by the notification of 6 cases amongst male students aged 18-28 years with dates of onset between 3 March and 21 March 1996. In response, a MR vaccination campaign was conducted to enhance population immunity to rubella within the university population and to reduce the likelihood of further cases. A total of 1795 students, staff and visitors were vaccinated. Vaccine coverage of 46% was estimated to be sufficient to boost rubella immunity in full time male students in university accommodation to 88.7-91.0%, just above the upper critical level of herd immunity for rubella of 85-88%. Students in colleges and universities in the UK will remain at increased risk of outbreaks of rubella and measles until the cohort who have received a two dose schedule of MR form the bulk of the college population. It may be prudent for tertiary education colleges and other institutions in the UK with young adults living in shared residential accommodation to offer MR vaccination to new entrants, targeting those who have not previously received the vaccine, between now and the year 2000. PMID- 9747768 TI - Indirect methods for estimating prevalent HIV infections: adults in England and Wales at the end of 1993. AB - Two indirect methods were used to estimate the point prevalence of HIV infection in England and Wales at the end of 1993 using data on diagnosed HIV infections, AIDS cases, HIV-related deaths and HIV testing behaviour from unlinked anonymous surveys. The methods estimated the proportion of all prevalent HIV infections that diagnosed infections represented. Most of those exposed to HIV infection through injecting drug use or sexual intercourse between men had had their infections diagnosed compared to less than half of those exposed through heterosexual intercourse. The total estimated number of prevalent infections was 22,350 for the diagnosis interval method and 20,540 for the test history method, and about 56-57% of these were in homo/bisexual men. These indirect methods are cheap and simple applications of surveillance data which provide estimates that compare favourably with those produced by more complex methods. PMID- 9747769 TI - Risk factors for cervical presence of human papillomavirus DNA among women at risk for HIV infection. DIANAIDS Collaborative Study Group. AB - Risk factors for cervical infection with human papillomavirus (HPV) were assessed among 236 Italian women at risk for human immunodeficiency virus (HIV) infection (intravenous drug users (IVDU) or sexual partners of males at risk for HIV infection). All study participants underwent a structured interview, determination of HIV serostatus and detection of HPV cervical infection by means of polymerase chain reaction (PCR). Overall, the cervical presence of HPV DNA was ascertained in 86 of these 236 women (36.4%), while squamous intraepithelial lesions (SIL) were diagnosed in 57 (24.1%). HPV-infected and non-infected women did not differ in age, education and cigarette smoking. A statistically significant trend in the risk of HPV infection with increasing number of lifetime sexual partners was noted (P = 0.01), but such trend was attenuated in multivariate analysis (multiple logistic regression (MLR) odds ratio (OR) for > or = 20 partners vs 1 = 1.6, 95% confidence intervals (CI): 0.4-5.9). A nearly threefold higher risk of HPV cervical infection emerged among IVDU women (MLR-OR: 2.7, 95% CI: 1.4-5.0), and this difference was not influenced by HIV serostatus. The prevalence of HIV infection was higher among HPV-positive than HPV-negative women (62.8% and 54.0%, respectively) (MLR-OR = 1.9, 95% CI: 0.9-3.8), and the proportion of women with less than 200 CD4+ cells/mm3 was slightly and not significantly higher among HPV-positive (47.1%) than negative women (37.2%). PMID- 9747770 TI - HBV and HCV infection among non-European Union immigrants in North-East Italy. AB - The status of hepatitis B virus (HBV) and hepatitis C virus (HCV) infection among non-European Union (non-EU) immigrants in North-East Italy was evaluated. Among the 1683 individuals tested the prevalence of HBsAg was 8.9% (150 subjects) and of HBV antibodies (anti-HBc with/without anti-HBs) was 38.9% (654 subjects). The distribution of HBV serological markers showed significant differences according to region of origin; the highest prevalence of infection (76.9%) and carriage (16.1%) was found in immigrants from sub-Saharan Africa. Among the 933 individuals screened for HCV infection, prevalence of antibody was much lower (0.9%) than that observed in the Italian general population (3.2-12.6%). The large number of HBV carriers among immigrants could increase the number of new adult infections due to life-style habits or professional risks in the host population. In contrast, the risk of HCV spread from non-EU immigrants is very low. PMID- 9747771 TI - Hepatitis B outbreak at Glenochil prison during January to June 1993. AB - This data linkage study examined the extent of hepatitis B transmission and co infection with HIV among 636 former inmates of Glenochil prison, Scotland, during an outbreak of bloodborne diseases in 1993 which was related to needle sharing. Eleven inmates imprisoned during the first half of 1993 presented with hepatitis B infection, of whom co-infection with HIV was detected in six. Based on dates of test results in relation to time of imprisonment, seven definitely acquired their hepatitis B infection within the prison. Only two infections were reported to Scotland's hepatitis B register and neither could be prison-linked. This outbreak of hepatitis B is the first of its kind to be reported but not the first to have occurred. It not only highlights the urgency for measures to prevent further spread of infection among prisoners but also illustrates the need for comprehensive surveillance of hepatitis B infection, and the need for a protocol on how to manage such outbreaks and on how to establish the extent of transmissions when acute hepatitis B occurs in prison. PMID- 9747772 TI - Absence of relationship between Schistosoma japonicum and hepatitis B virus infection in the Dongting lake region, China. AB - In order to determine whether infection with Schistosoma japonicum is related to a higher rate of infection with hepatitis B virus and/or to a higher probability of HBsAg chronic carriage, a population based survey was carried out in China in which HBV markers were studied in 112 subjects with long-lasting S. japonicum infection, and 93 subjects with no S. japonicum infection 37.5% of the cases and 40.9% of controls showed no markers of HBV infection. The prevalence rate of HBsAg was 12.5% in the cases and 12.9% in the controls. For anti-HBc and anti-HBs the figures were 59.8% and 59.8%, and 27.9% and 35.0%, respectively. These data do not support the hypothesis of an interaction between infection with hepatitis B virus and S. japonicum. PMID- 9747773 TI - Seasonality and factors associated with cryptosporidiosis among individuals with HIV infection. AB - The seasonality and factors associated with Cryptosporidium infection were assessed in a cohort of HIV-infected patients in Los Angeles County to better define the epidemiology of cryptosporidiosis among individuals with HIV. Data were analysed from a cohort of 4247 patients > or = 13 years of age with HIV infection enrolled from four outpatient facilities in Los Angeles, 1990-6. Cryptosporidiosis was diagnosed in 120 (2.8%) patients. Among the 1296 individuals with complete follow-up until death, cryptosporidiosis occurred in 69 (5.3%). The seasonal rate of cryptosporidiosis showed a modest bimodal trend with the highest rates occurring in March-May and September-October. There was no difference in the rate of cryptosporidiosis for the periods of heaviest rainfall (December-March) and low rainfall (April-November). Infection rates were higher among males (1.59 per 100 person-years) than females (0.92) and lower in blacks (0.98) than other racial/ethnic groups (1.80). A significant trend of decreasing cryptosporidiosis was observed with increasing age, with the highest rate (2.34) in the 13-34 year age group. A strong association between cryptosporidiosis and CD4+ count was noted. These data suggest that cryptosporidiosis among HIV infected individuals in Los Angeles County exhibits a modest spring and fall seasonality. This pattern of occurrence of cryptosporidiosis appears temporally unrelated to local rainfall patterns. Our findings suggest that HIV-infected men, individuals in younger age groups and those with CD4+ lymphocyte counts < 100 x 10(6)/l are at increased risk of cryptosporidiosis. Blacks with HIV infection appear less likely than other racial/ethnic groups to be diagnosed with Cryptosporidium infection. These results may provide insight into possible routes of transmission and sources of cryptosporidiosis infection in individuals with HIV. PMID- 9747774 TI - Comparisons of ELISA and Western blot assays for detection of Cryptosporidium antibody. AB - A seroprevalence survey was conducted using ELISA and Western blot (WB) assays for antibody to three Cryptosporidium antigens on 380 blood donors in Jackson County, Oregon. The purpose was to determine if either assay could detect serological evidence of an outbreak which occurred in Talent, Oregon 6 months earlier. The ELISA, which tested for combined IgG, IgA and IgM, and the WB, which tested separately for IgG and IgA, detected an almost twofold increase in serological response for persons who consumed Talent drinking water during the previous 11 months. The increases, however, were statistically significant (P < 0.05) only for the WB. The identification of serological evidence of infection, using sera collected 6 months after the end of the outbreak in a population not selected because of cryptosporidiosis-like illness, suggests that assays of Cryptosporidium-specific IgG and IgA may assist in estimating the magnitude of asymptomatic infections in the population. PMID- 9747775 TI - A two-year follow-up survey of antibody to Cryptosporidium in Jackson County, Oregon following an outbreak of waterborne disease. AB - To estimate the duration of Cryptosporidium-specific antibody, a Western blot assay measured antibody in paired sera from 124 residents of Jackson County, Oregon collected 0.5 and 2.5 years after the end of an outbreak in Talent, Jackson County. The outcome measure was the intensity of antibody responses, (which may approximate to a titre), to 27-kDa and 15/17-kDa antigens. Intensity of response to the 27-kDa antigen(s) declined to 54% of the 1992 value while responses to a 15/17-kDa antigen(s) remained close to the initial values. Increasing age of the donor predicted higher intensity of antibody to the 15/17 kDa antigen(s) in both the initial (P = 0.004) and follow-up (P = 0.038) surveys. No relationship was observed between age and antibody intensity for the 27-kDa antigen(s) during either survey (P > 0.10). Both the initial and follow-up surveys showed significant elevations in antibody intensity for Talent residents, possibly indicating a high endemic rate of infection/re-infection or high levels of chronic infection. PMID- 9747776 TI - Serpulina pilosicoli, waterbirds and water: potential sources of infection for humans and other animals. AB - Serpulina pilosicoli was isolated from 8 of 43 (19%) faecal specimens obtained from feral waterbirds sampled around a small lake at Perth Zoological Gardens, Western Australia, and from 3 of 7 (43%) samples of the lake water. The organism was only isolated from 1 of 204 (0.5%) samples from captive birds and animals in the zoological collection. Multilocus enzyme electrophoresis analysis of the isolates showed that they were genetically diverse, and none had identical electrophoretic profiles as those previously obtained from human beings, dogs, pigs and other avian species. To determine the survival time of S. pilosicoli in water, cells of strain 1648 were seeded into lake and tap water, and incubated at 4, 25 and 37 degrees C. The organism could be recultured from lake water for up to 66 days at 4 degrees C, and for 4 days at 25 degrees C. A healthy human volunteer who drank water seeded with S. pilosicoli strain Wes B became colonized, and developed abdominal discomfort and headaches. Contamination of water by faeces may represent a source of S. pilosicoli infection for both humans and animals. PMID- 9747777 TI - At-risk individuals in Feline Immunodeficiency Virus epidemiology: evidence from a multivariate approach in a natural population of domestic cats (Felis catus). AB - Prevalence of Feline Immunodeficiency Virus (FIV) infection was measured during 6 consecutive years in a natural rural population of domestic cats. Sex, age, weight, origin, group size and presence of antibodies to FIV were recorded for each sampled cat. Logistic regressions were used to estimate the influence of the recorded parameters on infection. FIV prevalence rates are as high as 19.6% in the total population, and do not statistically change between years, after controlling for changes in samples' age structure. FIV infection is characterized by risk factors linked to aggressive behaviour: old mature male adults having dispersed are more likely to be infected. A study of the cats group size and of the spatial distribution of infected individuals indicates the absence of infection clusters in males, and suggests the importance of roaming in the spreading of FIV. In conclusion, FIV infection spreads, with low contagiousness, mainly between particularly aggressive individuals, and the virus is endemic in this population. PMID- 9747778 TI - Pseudomonas aeruginosa as a cause of infectious diarrhoea. AB - Pseudomonas aeruginosa is not generally considered a cause of infectious diarrhoea. However, it was the predominant organism isolated from the faeces of 23 unrelated, hospital outpatients investigated in the course of a year for persistent (> 1 week duration) diarrhoea. To investigate the possible aetiological role of P. aeruginosa, these patient histories were reviewed and a selection of their faecal isolates were investigated in vitro (n > or = 10) and in vivo (n = 2) for virulence. The patients had a mean age of 60 years, were receiving antibiotics and/or had an underlying illness. Extensive microbiological investigations identified no other potential or recognized enteropathogen in the faeces of 20 of these patients. More than 40% of the isolates tested were able to adhere to HEp-2 cells and exhibited twitching motility (type IV pili), properties indicative of their ability to colonize the human intestine. Cytotoxic activity was demonstrated in bacterium-free cell supernatants of over 80% of isolates; supernatants of four isolates tested in infant mice were weakly enterotoxigenic. Two isolates intragastrically inoculated into clindamycin pre-treated rats established persistent infections and induced signs and symptoms of enteritis. Overall these findings suggest that P. aeruginosa can cause diarrhoea particularly in immunodeficient individuals. PMID- 9747779 TI - Analytic model to predict the strength of tendon repairs. AB - We developed an analytic model to predict suture load-sharing immediately after flexor tendon repair in the hand. Tendon repair was mathematically modeled as two nonlinear springs in parallel, representing separate core and peripheral sutures that were in series with a third nonlinear spring representing the tendon. To serve as a basis for, and validation of, our analytic model, fresh human flexor digitorum profundus tendons were harvested and mechanically tested either intact or after surgical repair in a variety of ways: core suture alone, superficial peripheral suture alone, deep peripheral suture alone, core suture plus superficial peripheral suture, and core suture plus deep peripheral suture. The stiffness and strength of the composite repairs predicted with use of the analytic model were comparable with those determined experimentally. Furthermore, the model predicted inequities in suture load-sharing, with 64% of the applied load carried by the peripheral suture when it was placed superficially, as compared with 77% when the peripheral suture was placed deep. Our results demonstrate a disparity in load-sharing within composite suture systems, the rectification of which may lead to significant improvement in the repair strength. To this end, we expect that our analytic model will serve as a basis for the design of more efficient, and consequently stronger, suture techniques. PMID- 9747780 TI - Use of mesenchymal stem cells in a collagen matrix for Achilles tendon repair. AB - This investigation tested the hypothesis that delivering mesenchymal stem cell seeded implants to a tendon gap model results in significantly improved repair biomechanics. Cultured, autologous, marrow-derived mesenchymal stem cells were suspended in a collagen gel delivery vehicle; the cell-gel composite was subsequently contracted onto a pretensioned suture. The resulting tissue prosthesis was then implanted into a 1-cm-long gap defect in the rabbit Achilles tendon. Identical procedures were performed on the contralateral tendon, but only the suture material was implanted. The tendon-implant constructs were evaluated 4, 8, and 12 weeks later by biomechanical and histological criteria. Significantly greater load-related structural and material properties were seen at all time intervals in the mesenchymal stem cell-treated tendons than in the contralateral, treated control repairs (p < 0.05), which contained suture alone with natural cell recruitment. The values were typically twice those for the control tissues at each time interval. Load-related material properties for the treated tissues also increased significantly over time (p < 0.05). The treated tissues had a significantly larger cross-sectional area (p < 0.05), and their collagen fibers appeared to be better aligned than those in the matched controls. The results indicate that delivering mesenchymal stem cell-contracted, organized collagen implants to large tendon defects can significantly improve the biomechanics, structure, and probably the function of the tendon after injury. PMID- 9747781 TI - Viability of fibroblast-seeded ligament analogs after autogenous implantation. AB - Fibroblast-seeded collagen scaffolds or ligament analogs are potentially useful for reconstruction of the anterior cruciate ligament of the knee. To provide lasting benefits, the seeded cells must survive implantation within the harsh synovial environment of the knee joint. Our objective was to determine the in vivo fate of autogenous fibroblast-seeded ligament analogs as a function of fibroblast source (anterior cruciate ligament or skin), implantation site (knee joint or subcutaneous space), and time after implantation (1, 2, 4, 6, or 8 weeks). Before implantation, fibroblasts were labeled with PKH26-GL, a fluorescent membrane dye. Immediately after retrieval of the implant, the viability of the labeled seeded cells was assessed under a fluorescent microscope. Viable seeded fibroblasts remained attached to the collagen fibers within the ligament analogs for at least 4 weeks (skin fibroblasts) or 6 weeks (anterior cruciate ligament fibroblasts) after implantation. A larger number of viable seeded cells were consistently observed in the subcutaneous space than in the knee joint. Scaffold resorption prevented observation at the 8-week time period. Fibroblast-seeded ligament analogs remained viable for prolonged periods in the knee joint and therefore have the potential to influence the formation and remodeling of neoligament tissue after reconstruction of the anterior cruciate ligament. PMID- 9747782 TI - Comparison of mRNA levels for matrix molecules in normal and disrupted human anterior cruciate ligaments using reverse transcription-polymerase chain reaction. AB - Midsubstance samples of anterior cruciate ligaments from seven normal human cadaver knees (16-50 years old) were harvested and compared with midsubstance pieces of scarred anterior cruciate ligaments from 30 patients (15-40 years old). RNA was isolated from each ligament, and the expression of type-I collagen, type III collagen, biglycan, decorin, lumican, and tissue inhibitor of metalloproteinase-1 was evaluated by quantitative reverse transcription polymerase chain reaction with use of beta-actin as the housekeeping gene. Data for injured ligaments were further compared statistically as a function of time after injury to better define patterns of cellular expression over time. Our hypothesis was that injured ligaments would show minimal cellular activity and decreasing activity over time. The results revealed that both normal and injured anterior cruciate ligaments contain cells that express mRNA for all molecules studied. However, cells in injured ligaments express much higher, but still proportional, quantities of message for type-I collagen and type-III collagen (p < 0.000001) and higher quantities of biglycan (p < 0.02) and tissue inhibitor of metalloproteinase-1 (p < 0.0003) than do cells in normal anterior cruciate ligaments. These levels remained elevated for longer than 1 year after injury. Linear regression analysis showed biglycan expression correlated with time from injury (r2 = -0.69; p = 0.007). These results collectively demonstrate that injured human anterior cruciate ligaments contain cells that express scar-like molecules and that the injured ligaments are likely continuing to remodel matrix over time. Furthermore, they suggest that human anterior cruciate ligaments have not failed to heal due to the failure of scar formation per se. The quality and quantity of this scar remain questionable; however, the possibility of its enhancement as a healing strategy for human anterior cruciate ligaments cannot be dismissed. PMID- 9747783 TI - Semiquantitative reverse transcription-polymerase chain reaction analysis of mRNA for growth factors and growth factor receptors from normal and healing rabbit medial collateral ligament tissue. AB - Growth factors and their receptors play an essential role in the development, maturation, and response to injury of all tissues. A number of studies have explored the possibility of improving ligament healing with exogenous growth factors. However, limited data is available regarding the endogenous growth factor network in ligaments on which any exogenous growth factors must impact. The purpose of this study was to assess the endogenous growth factor network with molecular techniques. By the sensitive reverse transcription-polymerase chain reaction technique, transcripts for a number of growth factors and receptors were detected with RNA isolated from normal and healing rabbit medial collateral ligament tissues. These include transforming growth factor-beta1, insulin-like growth factors I and II, basic fibroblast growth factor, endothelin-1, and the receptors for insulin and insulin-like growth factor II. Semiquantitative reverse transcription-polymerase chain reaction analysis of RNA from normal and scar tissues from the medial collateral ligament revealed that the levels of several transcripts were elevated in the scar tissue. It was not possible to confirm biological activity because of the hypocellularity of the tissues; however, the results obtained indicate that the reverse transcription-polymerase chain reaction approach to defining the endogenous growth factor-receptor phenotype is feasible, and further definition should contribute to the development of rational approaches to exogenous therapy to improve healing. PMID- 9747784 TI - Collagen fiber sliding during ligament growth and contracture. AB - It is hypothesized that the sliding of collagen fibers past one another plays an important role in changes of ligament length during growth or contracture. To explore this possibility, we used the fluorescent dye dichlorotriazinyl fluorescein to stain collagen fibers perpendicular to their orientation in a rat medial collateral ligament model. Growth, contracture, and control models (with rats weighing 50-75 g in the first and 500-600 g in the latter two groups) were studied. In the contracture model, the medial collateral ligament was transected distally. Marking sutures were used to verify the presence of growth or contracture in each medial collateral ligament. Fluorescence photomicrography after 2 weeks demonstrated stained collagen fibers protruding from either side of the original mark as one would expect, in either growth or contracture, if the fibers slid past one another and away from their initial location during changes in length. By measuring the initial and final widths of the growth and contracture model marks and correlating them to controls that had minimal growth (rats grow throughout their life) and were free of contracture, we have provided evidence that collagen sliding plays a significant role in changes in ligament length. PMID- 9747785 TI - Method to assess in vivo knee stability longitudinally in an animal model of ligament injury. AB - The purpose of this study was to develop a method to prospectively quantify passive knee stability in an animal model of joint injury over time. Knee stability is defined here as the amount of translation or rotation of the tibia relative to the femur for a given application of force or moment, respectively. Five animals that had undergone transection of the anterior cruciate ligament and three control animals that had undergone a sham operation were anaesthetized and positioned in a stereotaxic frame. Motion of the tibia relative to the femur was quantified with use of reflective markers secured to modified bone pins and a three-dimensional motion analysis system. External forces and moments in the transverse plane of the tibia were measured with use of force transducers based on a strain-gauge design. Longitudinal measurements of knee stability were made before either sham surgery (control animals) or transection of the ligament (experimental animals), immediately after surgery, and at 2 and 4 months after transection. The results showed that the animals tolerated the procedures well and that systematic measurements could be obtained. The method described here has the practical advantage over cross-sectional experimental designs in that the number of subjects can be decreased while maintaining statistical power and has the further conceptual advantage that individual changes can be accounted for over time. PMID- 9747786 TI - Longitudinal measurement of tibial motion relative to the femur during passive displacements in the cat before and after anterior cruciate ligament transection. AB - Passive anterior-posterior displacement and medial-lateral rotation of the tibia on the femur in the feline knee were assessed before transection of the anterior cruciate ligament, immediately after transection, and 2 and 4 months after transection. Four anaesthetized experimental and three sham-operated control animals were positioned in a stereotaxic frame. Motions of the tibia relative to the femur were measured with use of 60-Hz video motion analysis, while a strain gauged system allowed measurement of forces and moments applied to the tibia. Displacement at 15 N of anterior force and 30 degrees of knee flexion increased by an average of 6 mm following transection, and stiffness decreased by an average of 6 N/mm. At 2 and 4 months following transection, there were statistically significant reductions in this abnormal displacement. Stiffness during anterior displacement of the tibia at 30 degrees increased significantly from immediately after transection to 4 months. At 90 degrees, mean anterior displacement decreased from 5.1 mm immediately after transection to 2.9 mm at 4 months. Medial rotation at 30 degrees of knee flexion was significantly decreased from a mean of 16.5 degrees after transection to a mean of 10.7 degrees at 4 months. Changes in medial rotation at 90 degrees, lateral rotation at 90 degrees, and lateral rotation at 30 degrees were not statistically significant. These results indicate a significant change in secondary constraints to tibial motion in response to knee instability. PMID- 9747787 TI - In situ calibration of miniature sensors implanted into the anterior cruciate ligament part I: strain measurements. AB - The goals of this study were to (a) evaluate the differential variable reluctance transducer as an instrument for measuring tissue strain in the anteromedial band of the anterior cruciate ligament, (b) develop a series of calibration curves (for simple states of knee loading) from which resultant force in the ligament could be estimated from measured strain levels in the anteromedial band of the ligament, and (c) study the effects of knee flexion angle and mode of applied loading on output from the transducer. Thirteen fresh-frozen cadaveric knee specimens underwent mechanical isolation of a bone cap containing the tibial insertion of the anterior cruciate ligament and attachment of a load cell to measure resultant force in the ligament. The transducer (with barbed prongs) was inserted into the anteromedial band of the anterior cruciate ligament to record local elongation of the instrumented fibers as resultant force was generated in the ligament. A series of calibration curves (anteromedial bundle strain versus resultant force in the anterior cruciate ligament) were determined at selected knee flexion angles as external loads were applied to the knee. During passive knee extension, strain readings did not always follow the pattern of resultant force in the ligament; erratic strain readings were often measured beyond 20 degrees of flexion, where the anteromedial band was slack. For anterior tibial loading, the anteromedial band was a more active contributor to resultant ligament force beyond 45 degrees of flexion and was less active near full extension; mean resultant forces in the range of 150-200 N produced strain levels on the order of 3-4%. The anteromedial band was also active during application of internal tibial torque; mean resultant forces on the order of 180-220 N produced strains on the order of 2%. Resultant forces generated by varus moment were relatively low, and the anteromedial band was not always strained. Mean coefficients of variation for resultant force in the ligament (five repeated measurements) ranged between 0.038 and 0.111. Mean coefficients of variation for five repeated placements of the strain transducer in the same site ranged from 0.209 to 0.342. Insertion and removal of this transducer at the anteromedial band produced observable damage to the ligament. In our study, repeatable measurements were possible only if both prongs of the transducer were sutured to the ligament fibers. PMID- 9747788 TI - In situ calibration of miniature sensors implanted into the anterior cruciate ligament part II: force probe measurements. AB - The arthroscopically implantable force probe transducer, which measures the effects of local ligament fiber tension, was inserted into the anteromedial band of the anterior cruciate ligament after measurements with the differential variable reluctance transducer were completed in Part I of this study. The overall goals in Part II remained the same, with additional experiments included to determine the sensitivity of output voltage from the transducer to medial lateral placement of the device within the anteromedial band and to depth of placement within a given insertion hole. Calibration curves of output voltage from the arthroscopically implantable force probe transducer versus resultant force in the ligament were generated during a separate series of knee-loading experiments identical to those performed in Part I. The output voltage for a given probe placement was highly sensitive to the depth of implantation into the anteromedial band. When the probe was completely buried within the ligament, voltage outputs were often sporadic or absent even though surface fibers had clearly developed tension. When the probe was only partially inserted into the hole, such that the end of the probe was slightly proud to the surface, voltage output was significantly higher as the device measured tension in the superficial fibers. Voltage outputs for proud placement were always significantly higher than corresponding voltages for deep placements for all test conditions. With proud placements, voltage outputs were not sensitive to small deviations in medial lateral position within the anteromedial band. Mean coefficients of variation for output voltage (four repeated placements of the probe into the same central hole) ranged from 0.156 to 0.359 (deep and proud insertions). Output voltage from the probe generally followed the pattern of resultant force in the ligament during passive knee extension. For anterior tibial loading, the contribution of deep fibers to resultant force did not depend on the knee flexion angle at which the test was conducted; the contribution of superficial fibers was greatest beyond 45 degrees of flexion and least at full extension. The contributions of the anteromedial band to resultant force in the ligament were not significantly different between the three modes of loading (anterior tibial force, internal tibia torque, and varus moment) at either 0 or 10 degrees of flexion; this was true for both superficial and deep fibers. We found it necessary to secure the probe within the insertion site using a suture (for both deep and proud placements) to obtain repeatable readings. Puncturing the anteromedial band clearly produced tissue damage; the insertion hole often produced a permanent plane of cleavage in the anteromedial band. However, this tissue damage did not alter the overall ability of the ligament to generate resultant force. PMID- 9747789 TI - Mechanical properties of wrist extensor tendons are altered by the presence of rheumatoid arthritis. AB - The in vitro mechanical properties of 14 wrist extensor tendons salvaged at surgery from patients with inflammatory (rheumatoid) arthritis and noninflammatory arthrosis were measured in uniaxial tension and compared. The rheumatoid tendons had higher extensibility at low stresses, lower stiffness in the linear portion of the stress-strain curve, greater rates of stress relaxation, and lower ultimate strengths than did the nonrheumatoid tendons. Differences in tangent modulus, stress remaining at 100 seconds, and ultimate tensile strength were significant at the 95% confidence level. In vivo, mechanically impaired tendons may play an important role in destabilization of the wrist in patients with rheumatoid arthritis. PMID- 9747790 TI - Local tissue properties in bone healing: influence of size and stability of the osteotomy gap. AB - To characterize the site-specific mechanical and histological properties in fracture repair and to relate these properties to the initial mechanical situation, an experimental fracture model was used in the metatarsus of 42 sheep. The mechanical situation of a transverse osteotomy was described by three gap sizes (1, 2, or 6 mm) and two amounts of strain (7 or 31%). An external fixator that allowed a defined axial movement provided control of these settings. Nine weeks following surgery, the healing area was dissected and tensile and compressive properties were measured in subregions of the fracture gap and the periosteal callus. The central, sagittal section was used for quantitative histology. We found the quality of the tissue along the osteotomy line to be most important for regaining mechanical stability. Increasing the size of osteotomy gaps resulted in poorer mechanical and histological qualities, and the repair process was less complete. Interfragmentary strain did not significantly influence the repair process. The smaller strain levels had already stimulated the secondary repair process, and this stimulatory effect could not be further enhanced by increasing the amount of strain. Our finding that large gaps between bone segments were not as well healed as were smaller gaps suggests that it is advantageous to avoid large gaps in fracture treatment. PMID- 9747791 TI - Nonlinear dependence of loading intensity and cycle number in the maintenance of bone mass and morphology. AB - The daily stress stimulus theory of bone adaptation was formulated to describe the loading conditions necessary to maintain bone mass. This theory identifies stress/strain magnitude and loading cycle number as sufficient to define an appropriate maintenance loading signal. Here, we extend the range over which loading cycle number has been evaluated to determine whether the daily stress stimulus theory can be applied to conditions of very high numbers of loading cycles at very low strain magnitudes. The ability of a relatively high-frequency (30-Hz) and moderate-duration (60-minute) loading regimen to maintain bone mass in a turkey ulna model of disuse osteopenia was evaluated by correlating the applied strain distributions to site-specific remodeling activity. Changes in morphology were investigated following 8 weeks of disuse compared with disuse plus daily exposure to 108,000 applied loading cycles sufficient to induce peak strains of approximately 100 microstrain. A strong correlation was observed between the preservation of bone mass and longitudinal normal strain (R = 0.91) (p < 0.01). The results confirm the strong antiresorptive influence of mechanical loading and identify a threshold near 70 microstrain for a daily loading cycle regimen of approximately 100,000 strain cycles. These results are not consistent with the daily stress stimulus theory and suggest that the frequency or strain rate associated with the loading stimulus must also play a critical role in the mechanism by which bone responds to mechanical strain. PMID- 9747792 TI - Effects of injurious compression on matrix turnover around individual cells in calf articular cartilage explants. AB - The effects of mechanical injury on the metabolism of cartilage matrix are of interest for understanding the pathogenesis of osteoarthrosis and the development of strategies for cartilage repair. The purpose of the present study was to examine the effects of injury on matrix turnover in a calf articular cartilage explant system for which the effects of mechanical loading on cell activity and the cell-mediated pathways of matrix metabolism are already well characterized. New methods of quantitative autoradiography were used in combination with established biochemical and biomechanical techniques for the analysis of cell and matrix responses to acute mechanical injury, with particular attention to the processes of localized matrix turnover in the cell-associated matrices of individual chondrocytes. Matrix deposition and turnover around cells in control explants was spatially dependent, with the highest rates of proteoglycan deposition and turnover and the lowest rates of collagen deposition (as indicated by [3H]proline autoradiography) occurring in the pericellular matrix. Injurious compression was associated with (a) an abrupt decrease in the tensile load carrying capacity of the collagen matrix, apparently associated with mechanical failure of the tissue, (b) a considerable but subtotal decrease in cell viability, marked by the emergence of an apparently inactive cell population interspersed within catabolically active but abnormally large cells, and (c) sustained, elevated rates of proteoglycan turnover, particularly in the cell associated matrices of apparently viable cells, which involved the increased release of aggregating species in addition to a spectrum of degradation fragments that were also in controls. These results may represent an in vitro model for the responses of chondrocytes and the cartilage extracellular matrix to mechanical injury. PMID- 9747793 TI - Changes in cell, matrix compartment, and fibrillar collagen volumes between growth-plate zones. AB - To define the contributions of changes in cell, matrix compartment, and fibrillar collagen volumes to longitudinal bone growth, we measured the differences in cell, pericellular/territorial matrix and interterritorial matrix volumes, and fibrillar collagen concentrations between the upper proliferative and lower hypertrophic zones of the proximal tibial physes of six miniature pigs. The mean numerical density of cells decreased from 110,000 cells/mm3 in the upper proliferative zone to 59,900 cells/mm3 in the lower hypertrophic zone. The mean cell volume increased nearly 5-fold (from 1,174 to 5,530 microm3), and the total matrix volume per cell increased 46% (from 8,040 to 11,760 microm3/cell) between the upper proliferative and lower hypertrophic zones. Both the pericellular/territorial matrix volume per cell and the interterritorial matrix volume per cell increased between the upper proliferative and lower hypertrophic zones; the pericellular/territorial matrix volume per cell increased 61% (from 4,580 to 7,390 microm3/cell), whereas the interterritorial matrix volume per cell increased 26% (from 3,460 to 4,370 microm3/cell). The total increase in mean cell volume of 4,356 microm3 exceeded the total increase in mean matrix volume per cell of 3,720 microm3; the total mean pericellular/territorial matrix volume per cell increased more than the total mean interterritorial matrix volume per cell (2,810 compared with 910 microm3/cell). Fibrillar collagen concentration was greater in the interterritorial matrix than in the pericellular/territorial matrix in both zones and increased in both matrix compartments between the upper proliferative and lower hypertrophic zones. The amount of fibrillar collagen per cell also increased in both matrix compartments between the upper proliferative and lower hypertrophic zones (from 1,720 to 3,100 microm3/cell in the pericellular/territorial matrix and from 1,490 to 2,230 microm3/cell in the interterritorial matrix; thus, the total amount of fibrillar collagen per cell increased from 3,210 to 5,530 microm3/cell). Growth rate was inversely related to the cell numerical density in the upper proliferative and lower hypertrophic zones and was directly related to interterritorial matrix volume per cell in the upper proliferative zone and to pericellular/territorial matrix volume per cell in the lower hypertrophic zone. These results show that cell enlargement contributes more to longitudinal bone growth than does increased matrix volume, that increased pericellular/territorial matrix volume makes a greater contribution to growth than does increased interterritorial matrix volume, and that the total amount of fibrillar collagen per cell increases between the upper proliferative and lower hypertrophic zones. The differences between the two matrix compartments in increase in volume, fibrillar collagen concentration, and amount of fibrillar collagen per cell strongly suggest that they differ not only in matrix organization but in rate of matrix accumulation and assembly and that these differences give the two compartments different roles in skeletal growth. PMID- 9747794 TI - Effect of ciprofloxacin on the proliferation of osteoblast-like MG-63 human osteosarcoma cells in vitro. AB - Locally applied antibiotic therapy is gaining popularity for the treatment of infections associated with open fractures and posttraumatic osteomyelitis. With use of local techniques, ciprofloxacin levels as high as 1,300 microg/ml, or over 200 times the bone levels achieved with intravenous administration, have been reported. To study the possible effects of ciprofloxacin on bone, osteoblast-like cells from the MG-63 human osteosarcoma cell line were studied. The cells were grown in antibiotic-free media and exposed to concentrations of ciprofloxacin at 0, 10, 100, 200, and 1,000 microg/ml to establish an initial dose-response curve. Media containing the appropriate dose of ciprofloxacin were changed every 24 hours. Cell number and [3H]thymidine incorporation per cell were determined at 0, 24, and 72 hours. A second dose-response curve was performed at concentrations of 0, 10, 20, 40, and 80 microg/ml. Three experiments, each with four observations, were performed. The results of this study demonstrated that ciprofloxacin caused significant decreases (p < 0.05) in cell number at 40 microg/ml at 24 hours and 20 microg/ml at 72 hours. [3H]thymidine incorporation per cell decreased significantly at levels of 80 microg/ml at 24 hours and 20 microg/ml at 72 hours. The authors conclude that reported local levels of ciprofloxacin seen in vivo inhibit the proliferation of human osteoblast-like cells in vitro. PMID- 9747795 TI - Analysis of optimal range of socket orientations in total hip arthroplasty with use of computer-aided design simulation. AB - A three-dimensional computer-aided design model of a total hip replacement was used to study the effects of anteversion and abduction of the acetabular component and anteversion and varus-valgus angulation of the femoral component on the range of hip flexion and extension that could be obtained without component impingement. Impingement of the component was defined as impingement between the neck of the femoral component and the edge of the acetabular component. To achieve an angle of hip flexion greater than 90 degrees and an extension angle greater than 30 degrees without component impingement, the optimal angulations were found to be between 1 and 30 degrees of anteversion and 30 and 50 degrees of abduction of the acetabular component, as well as 10 degrees of anteversion of the femoral component. When the valgus angulation of the femoral component was reduced from 7 to 0 degrees, the allowable range of flexion without impingement increased under the same conditions of acetabular-component orientation and femoral-component anteversion. Significant inverse correlations were found between the anteversion angle of the acetabular component and both the lumbar lordosis angle and the sacrohorizontal angle. PMID- 9747796 TI - Lengths of hamstrings and psoas muscles during crouch gait: effects of femoral anteversion. PMID- 9747797 TI - Substrate profiles and expression of caffeoyl coenzyme A and caffeic acid O methyltransferases in secondary xylem of aspen during seasonal development. AB - Seasonal expression of caffeoyl-CoA O-methyltransferase (EC 2.1.1.104) was analyzed in aspen developing secondary xylem in parallel with caffeate O methyltransferase (EC 2.1.1.68). Enzyme activity and mRNA levels for both enzymes peaked in the middle of the growing season. These results strongly suggest that both forms of O-methyltransferase were actively participating in lignin precursor biosynthesis during the growing season. To determine the role of each enzyme form, xylem extracts from two days in the growing season were assayed with four substrates: caffeoyl-CoA, 5-hydroxyferuloyl-CoA, caffeate acid and 5 hydroxyferulic acid. Recombinant forms of caffeoyl-CoA and caffeate O methyltransferase were also assayed with these substrates. The recombinant enzymes have different substrate specificity with the caffeoyl-CoA O methyltransferase being essentially specific for CoA ester substrates with a preference for caffeoyl-CoA, while caffeate O-methyltransferase utilized all four substrates with a preference for the free acid forms. We suggest that caffeoyl CoA O-methyltransferase is likely to be responsible for biosynthesis of lignin precursors in the guaiacyl pathway and may represent a more primitive enzyme form leftover from very early land plant evolution. Caffeate O-methyltransferase is more likely to be responsible for lignin precursor biosynthesis in the syringyl pathway, especially since it can catalyze methylation of 5-hydroxyferuloyl-CoA quite effectively. This latter enzyme form then may be considered a more recently evolved component of the lignin biosynthetic pathways of the evolutionarily advanced plants such as angiosperms. PMID- 9747798 TI - Insertional inactivation of the tomato polygalacturonase gene. AB - The site-selected insertion (SSI) procedure was used to generate insertional knockout mutations in the gene for tomato polygalacturonase (PG), a critical enzyme in fruit ripening. Previously, it had been shown that the Dissociation (Ds) elements in a select group of tomato plants frequently inserted into PG, at least in somatic tissues. DNA isolated from pollen produced by progeny of these plants was screened by SSI to identify plants likely to transmit the insertions in PG to progeny. These results identified one family as likely candidate for yielding germinally transmitted insertions. Four thousand progeny were screened and five were found containing germinally transmitted Ds insertions in PG, one of which contained two Ds insertions in PG. The Ds elements were stabilized by genetically removing the transposase and four of the five insertions were recovered as homozygous in the next generation. Enzymatic analysis of fruit from these individuals demonstrated that there was at least a 1000-fold reduction in polygalacturonase levels in those plants bearing Ds insertions in PG exons. Individuals with modified PG sequences due to the sequence footprint, resulting from excision of the element, were identified using the single-strand conformational polymorphism (SSCP) method. Enzymatic analysis of fruit from a plant homozygous for one such excision allele showed a significant reduction in polygalacturonase activity. Since there is no transgenic material left in PG, this demonstrates the ability to modify a gene of commercial value in planta and subsequently removing all transgenic material. PMID- 9747799 TI - Cloning and characterization of cold-regulated glycine-rich RNA-binding protein genes from leafy spurge (Euphorbia esula L.) and comparison to heterologous genomic clones. AB - Leafy spurge (Euphorbia esula) is a perennial weed which is capable of acclimating to sub-freezing temperatures. We have used the differential display technique to identify and clone a cDNA for a cold-regulated gene (cor20) which hybridizes to mRNAs that accumulate specifically during the cold acclamation process. The cor20 cDNA was used to isolate two different genomic clones. Both clones were similar but not identical to each other and the cDNA. Sequence analysis of the genomic clones indicated that they share considerable homology to a group of glycine-rich RNA-binding protein genes. Comparison of the promoter region from the three clones (Ccr1 from Arabidopsis. BnGRP10 from Brassica napus, and GRRBP2 from Euphorbia esula) have identified at least two conserved motifs. CAGC is most likely involved in cold regulation and AACCCYAGTTA, is conserved but has no known function. RNAs which hybridize to cor20 reach maximal expression in less than 2 days after exposure of the plant to temperatures of 5 degrees C, and remains at high levels in the plant for at least 30 days so long as the plant is left in the cold. These RNAs drop to control levels within 24 h when the plant is returned to normal growing temperatures. Transcripts which hybridize to cor20 do not accumulate under conditions of drought or heat stress. These transcripts are induced in response to low temperatures in roots, stems and leaves, but are expressed constitutively in tissue culture at control temperatures. PMID- 9747800 TI - Overexpression of deltaEmBP, a truncated dominant negative version of the wheat G box binding protein EmBP-1, alters vegetative development in transgenic tobacco. AB - As a first step toward elucidating the in vivo function of plant bZIP proteins and their related G-box cis elements, we have introduced a dominant negative inhibitor of G-box-dependent transcriptional activation into tobacco plants by transforming them with a truncated EmBP-1 gene (deltaEmBP) containing the DNA binding and dimerization domains under the control of the CaMV 35S promoter. Five independent lines of transgenic plants expressing deltaEmBP were identified, as demonstrated by immunodetection of the transgenic protein in leaf extracts, and the ability of the protein to bind a target G-box DNA sequence. The transgenic plants exhibited an abnormal phenotype characterized by interveinal chlorosis, growth inhibition and weakening of stems and petioles, the severity of which positively correlated with deltaEmBP expression and G-box DNA binding capability. Furthermore, development of chlorosis and growth inhibition was dependent on growth irradiance. Low light promoted the development of interveinal chlorosis and growth inhibition in the transgenic plants, whereas high light conditions led to near-complete amelioration of the abnormal phenotype. Transgenic plants under both light regimes showed signs of impaired stem and petiole function which was not observed in wild-type tobacco. RhcS gene expression was not significantly altered by deltaEmBP expression, suggesting that down-regulation of this gene was not responsible for the altered phenotype. The results suggest that G-box elements specific for the EmBP-1 class of bZIP proteins have an important developmental function in vegetative plant tissues, and that the trans-dominant negative mutant approach is a useful tool for continued in vivo functional analysis of bZIP transcription factors and their corresponding cis elements in plants. PMID- 9747801 TI - Identification of promoter elements in a low-temperature-responsive gene (blt4.9) from barley (Hordeum vulgare L.). AB - The blt4 barley gene family encodes non-specific lipid transfer proteins and has been shown, by in situ localisation, to be expressed in the epidermal cells of leaves. The transcriptionally controlled, low-temperature-responsive member of this gene family, blt4.9, is predominantly expressed in shoot meristems. The promoter region (1938 bp) of blt4.9 contains sequence motifs which have been implicated in responses to low temperature, abscisic acid and other environmental factors. Deletion analysis showed that a 42 bp sequence proximal to, but not including, the CAAT and TATA boxes, confers enhanced low-temperature response to a reporter gene in a barley shoot explant transient expression system. Other promoter regions were shown to contain negative and positive regulatory regions. Electrophoretic mobility shift analysis (EMSA) was used with nuclear proteins from either low-temperature- or control-temperature-treated plants to further investigate the blt4.9 promoter. Synthetic oligonucleotides were used to identify a hexanucleotide, CCGAAA, within the 42 bp, low-temperature-responsive promoter region, as the binding site of a low-mobility nuclear protein complex. This complex was present in nuclear extracts from both low-temperature-treated and control plants and was the only complex formed within this region. Mutation of the CCGAAA motif within the low-temperature-responsive 42 bp promoter sequence reduced low-temperature responsiveness to basal levels. A related upstream element, CCGAC, known to be a low-temperature-responsive element in other plants, did not bind to nuclear proteins in this study. It is proposed that the hexanucleotide CCGAAA, at -195 from the first ATG, is involved in the low temperature response of blt4.9 in barley. PMID- 9747802 TI - Arabidopsis thaliana vegetative storage protein (VSP) genes: gene organization and tissue-specific expression. AB - We have previously identified two cDNAs encoding vegetative storage proteins (VSPs) in Arabidopsis thaliana. Unlike soybean in which VSPs accumulate at high levels in leaves, A. thaliana VSP mRNAs are abundant in flowers. To understand tissue-specific expression and possible roles of VSPs on reproductive organ development, genes corresponding to VSPs (Vsp1 and Vsp2) and their putative promoters were characterized in this study. Genomic sequence analysis revealed that Vsp1 and Vsp2 resemble each other except in their introns, and that these two genes were organized in a tandem array with an interval of 6 kb in a region. The expression patterns of Vsp1 and Vsp2 were examined using transgenic A. thaliana plants carrying a promoter from Vsp1 or Vsp2 fused to a bacterial beta glucuronidase (GUS) reporter gene. The promoter from Vsp1 expressed its effect in gynoecia, especially in styles, the basal and distal ends of ovaries and in siliques, whereas the promoter from Vsp2 showed its activity in vegetative shoots, petioles, peduncles and receptacles of floral organs. These results suggest that expression of Vsp1 and Vsp2 may be developmentally regulated in A. thaliana. In the transgenic plants, the GUS activity was induced by wounding in an area around the mid-rib of leaves. Therefore, Vsp1 and Vsp2 promoters appear to have elements required for both tissue specificity and wounding. PMID- 9747803 TI - Characterization of the rice pathogen-related protein Rir1a and regulation of the corresponding gene. AB - In rice (Oryza sativa L.), local acquired resistance against Pyricularia oryzae (Cav.), the causal agent of rice blast, can be induced by a preinoculation with the non-host pathogen Pseudomonas syringae pv. syringae. We have cloned a cDNA (Rir1a) and a closely related gene (Rir1b) corresponding to transcripts that accumulate in leaf tissue upon inoculation with P. syringae pv. syringae. The cDNA encodes a putative 107 amino acid protein, Rir1a, that exhibits a putative signal peptide cleavage site in its hydrophobic N-terminal part and a C-terminal part that is relatively rich in glycine and proline. The Rir1b gene contains a Tourist and a Wanderer miniature transposable element in its single intron and encodes a nearly identical protein. Rir1a is similar in sequence (ca. 35% identical and ca. 60% conservatively changed amino acids) to the putative Wir1 family of proteins that are encoded by pathogen-induced transcripts in wheat. Using antibodies raised against a Rir1a-fusion protein we show that Rir1a is secreted from rice protoplasts transiently expressing a 35S::Rir1a construct and that the protein accumulates in the cell wall compartment of rice leaves upon inoculation with P. syringae pv. syringae. Possible roles of Rir1a in pathogen defense are discussed. PMID- 9747804 TI - Water-deficit-responsive proteins in maritime pine. AB - Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) was used to identify drought-responsive proteins during progressive water deprivation of two-year old maritime pine seedlings. Stress was applied by withholding water during vegetative growth. Needles were sampled before, during and after the stress. Out of about 1000 spots that were quantified by computer analysis, 38 responded during stress. Some proteins were accumulated while others were suppressed. One to three internal microsequences were obtained for 11 proteins, 10 of which were identified on the basis of sequence homologies. These proteins are quite diverse and are involved in photosynthesis, cell elongation, antioxidant metabolism and lignification. PMID- 9747805 TI - Irregular patterns of transgene silencing in allohexaploid oat. AB - An irregular pattern of transgene silencing was revealed in expression and inheritance studies conducted over multiple generations following transgene introduction by microprojectile bombardment of allohexaploid cultivated oat (Avena sativa L.). Expression of two transgenes, bar and uidA, delivered on the same plasmid was investigated in 23 transgenic oat lines. Twenty-one transgenic lines, each derived from an independently selected transformed tissue culture, showed expression of both bar and uidA while two lines expressed only bar. The relationship of the transgenic phenotypes to the presence of the transgenes in the study was determined using (1) phenotypic scoring combined with Southern blot analyses of progeny, (2) coexpression of the two transgenic phenotypes since the two transgenes always cosegregated, and (3) reactivation of a transgenic phenotype in self-pollinated progenies of transgenic plants that did not exhibit a transgenic phenotype. Transgene silencing was observed in 19 of the 23 transgenic lines and resulted in distorted segregation of transgenic phenotypes in 10 lines. Silencing and inheritance distortions were irregular and unpredictable. They were often reversible in a subsequent generation of self pollinated progeny and abnormally segregating progenies were as likely to trace back to parents that exhibited normal segregation in a previous generation as to parents showing segregation distortions. Possible causes of the irregular patterns of transgene silencing are discussed. PMID- 9747806 TI - The Arabidopsis Athb-8, -9 and -14 genes are members of a small gene family coding for highly related HD-ZIP proteins. AB - We report the isolation and characterization of two Arabidopsis homeobox genes highly related to the Athb-8 gene. The full-length cDNAs encode proteins of 841 and 852 amino acids which we have designated Athb-9 and -14, respectively. Athb 8, -9 and -14 are members of a small family of HD-Zip proteins (HD-ZIP III) characterized by a HD-Zip motif confined to the N-terminus of the polypeptide. The spatial organization of the HD-Zip domain of Athb-8, -9 and -14 is different from that of the Athb-1 (a member of the HD-ZIP I family) and Athb-2 (a member of the HD-ZIP II family) HD-Zip domains. DNA binding analysis performed with random sequence DNA templates showed that the Athb-9 HD-Zip (HD-Zip-9) domain, but not the Athb-9 HD alone, binds to DNA. The HD-Zip-9 domain recognizes a 11 bp pseudopalindromic sequence (GTAAT(G/C)ATTAC), as determined by selecting high affinity binding sites from random-sequence DNA. Moreover, gel retardation assays demonstrated that the HD-Zip-9 domain binds to DNA as a dimer. These data support the notion that the HD-ZIP III domain interacts with DNA recognition elements in a fashion similar to the HD-ZIP I and II domains. PMID- 9747808 TI - Identification of preferred binding sites of a light-inducible DNA-binding factor (MNF1) within 5'-upstream sequence of C4-type phosphoenolpyruvate carboxylase gene in maize. AB - MNF1 is a factor which specifically binds to a 318 bp fragment (-1012 to -695) in the 5'-flanking region of the C4-type phosphoenolpyruvate carboxylase gene in Zea mays (Yanagisawa et al., Mol Gen Genet 224 (1990) 325-332). The most preferred binding site of MNF1 determined by a 2 bp mutation-scanning assay was an octamer sequence, GTGCCCTT, which is located within the repeated sequences (RS1; -886 to 849, -846 to -807). Furthermore, a PCR-mediated selection-amplification assay identified both the octamer sequence, GTGCCC(A/T)(A/T), and an additional sequence. CC(G/A)CCC, the latter of which was similar to the Sp1 sites in vertebrates. Specific binding of MNF1 to each of the supposed binding sites was confirmed with double-stranded monomers as probes. Considering native molecular mass of MNF1 (ca. 500 kDa), a protein complex is expected. In addition, MNF1 is anticipated to have two distinct DNA-binding proteins since the MNF1 binding to CCGCCC element was 1,10-phenanthroline-dependent whereas the MNF1 binding to the octamer was independent. Wide distribution of the MNF1 binding sequences within the 1 kb promoter region accounts for broad interactions of MNF1. Moreover, specific DNA binding due to MNF1, which was not observed in the nuclear extract derived from germinated and cultivated plants in darkness, appeared after a white light pulse. This finding suggests the involvement of the protein complex in the light-dependent transcriptional control in the gene expression. PMID- 9747807 TI - Induction of mRNA accumulation corresponding to a gene encoding a cell wall hydroxyproline-rich glycoprotein by fungal elicitors. AB - The Hrgp (hydroxyproline-rich glycoprotein) gene codes in maize for one of the most abundant proteins of the cell wall. HRGPs may contribute to the structural support of the wall and they have also been involved in plant defense mechanisms. This second aspect has been tested for the Hrgp gene in maize where, in contrast with the situation in dicot species, the gene is encoded by a single-copy sequence. Hrgp mRNA accumulation is induced in maize suspension-cultured cells by elicitors, isolated either from maize pathogenic or non-pathogenic fungi. The induction of Hrgp mRNA accumulation by elicitor extracted from Fusarium moniliforme has been studied in detail. The level of induction depends on elicitor concentration and remains high until at least 24 h. Ethylene and protein phosphorylation appear to be involved in the transduction pathway of Hrgp gene activation by the F. moniliforme elicitor but not by 5 microM methyl jasmonate or 1 mM salycilic acid. Different compounds known to participate in plant stress responses such as ascorbic acid or reduced glutathione have also a positive effect on Hrgp mRNA accumulation. PMID- 9747809 TI - Wheat DNA polymerase CI: a homologue of rat DNA polymerase beta. AB - We have previously described a low-molecular-weight DNA polymerase (52 kDa) from wheat embryo: DNA polymerase CI (pol CI). This enzyme shares some biochemical properties with animal DNA polymerase beta (pol beta). In this report, we analyse pol CI in wheat embryo germination. Immunodetection and measurement of the enzyme activity show that wheat pol CI remains at a constant level during germination, whereas dramatic changes of the replicative DNA polymerase A and B activities were previously reported. We observe that the level of pol CI in physiological conditions (embryo germination and dividing cell culture) is in agreement with a pol beta-type DNA polymerase. By microsequencing of the electroblotted 52 kDa polypeptide, we determined the sequence of a dodecapeptide from the N-terminal region. A comparative analysis of the N-terminal pol CI peptide with some mammalian pol beta sequences shows a clear homology with helix 1 of the N terminal ssDNA domain (residues 15 to 26) of the rat pol beta. Thus, the helical structure of this region should be conserved in the wheat peptide. This represents the first evidence of a partial primary structure of a beta-type DNA polymerase in plants. PMID- 9747810 TI - Identification of tomato Lhc promoter regions necessary for circadian expression. AB - Expression of the light-harvesting complex protein genes (Lhc) is under the control of a circadian clock. To dissect the molecular regulatory components of the circadian clock a promoter deletion analysis of four tomato Lhc genes was performed in transgenic tobacco plants. The important 5'-upstream promoter regions are present at different positions relative to the transcription start site of Lhc b1*1, b1*2, Lhc a3 and Lhc a4. A short sequence of 47 nucleotides is necessary for conferring circadian Lhc mRNA oscillations. Sequence alignment of the specified promoter regions revealed a novel motif 'CAANNNNATC'. This motif is conserved in 5'-upstream regions of clock controlled Lhc genes and overlaps with a sequence relevant in phytochrome mediated gene expression. PMID- 9747811 TI - A monocot pollen-specific promoter contains separable pollen-specific and quantitative elements. AB - The region of the promoter of the pollen-specific maize gene, ZM13, from -119 to 37 was analyzed by a linker-scanning type of substitution mutagenesis and two areas were shown to be responsible for pollen expression: a proximal region delineated by mutations from -84 to -53 that conferred pollen specificity, and an upstream region delineated by a mutation from -107 to -102 (Q-element) that could increase the expression of the proximal region but showed no ability to cause expression in pollen on its own. Replacement of both of these areas with other sequences including the CaMV 35S promoter failed to replace activity. PMID- 9747812 TI - Cloning and characterization of two maize cDNAs encoding cinnamoyl-CoA reductase (CCR) and differential expression of the corresponding genes. AB - Cinnamoyl-CoA Reductase (CCR, EC 1.2.1.44) catalyses the first step of the lignin pathway. Two full-length cDNAs identified by sequence analysis as CCR-encoding cDNAs were isolated from a maize root cDNA library. These two cDNAs designated ZmCCR1 and ZmCCR2 exhibit 73% sequence conservation at the nucleotide level for their coding regions and are relatively divergent at their 5'- and 3' untranslated regions. They both contain a common signature which is thought to be involved in the catalytic site of CCR. Northern blot analysis indicated that ZmCCR2 was expressed at very low levels in roots whereas ZmCCR1 was widely expressed in different organs. The high level of ZmCCR1 gene expression along the stalk suggests that the corresponding enzyme is probably involved in constitutive lignification. PMID- 9747813 TI - Radiation pneumonitis as a function of mean lung dose: an analysis of pooled data of 540 patients. AB - PURPOSE: To determine the relation between the incidence of radiation pneumonitis and the three-dimensional dose distribution in the lung. METHODS AND MATERIALS: In five institutions, the incidence of radiation pneumonitis was evaluated in 540 patients. The patients were divided into two groups: a Lung group, consisting of 399 patients with lung cancer and 1 esophagus cancer patient and a Lymph./Breast group with 78 patients treated for malignant lymphoma, 59 for breast cancer, and 3 for other tumor types. The dose per fraction varied between 1.0 and 2.7 Gy and the prescribed total dose between 20 and 92 Gy. Three-dimensional dose calculations were performed with tissue density inhomogeneity correction. The physical dose distribution was converted into the biologically equivalent dose distribution given in fractions of 2 Gy, the normalized total dose (NTD) distribution, by using the linear quadratic model with an alpha/beta ratio of 2.5 and 3.0 Gy. Dose-volume histograms (DVHs) were calculated considering both lungs as one organ and from these DVHs the mean (biological) lung dose, NTDmean, was obtained. Radiation pneumonitis was scored as a complication when the pneumonitis grade was grade 2 (steroids needed for medical treatment) or higher. For statistical analysis the conventional normal tissue complication probability (NTCP) model of Lyman (with n=1) was applied along with an institutional dependent offset parameter to account for systematic differences in scoring patients at different institutions. RESULTS: The mean lung dose, NTDmean, ranged from 0 to 34 Gy and 73 of the 540 patients experienced pneumonitis, grade 2 or higher. In all centers, an increasing pneumonitis rate was observed with increasing NTDmean. The data were fitted to the Lyman model with NTD50=31.8 Gy and m=0.43, assuming that for all patients the same parameter values could be used. However, in the low dose range at an NTDmean between 4 and 16 Gy, the observed pneumonitis incidence in the Lung group (10%) was significantly (p=0.02) higher than in the Lymph./Breast group (1.4%). Moreover, between the Lung groups of different institutions, also significant (p=0.04) differences were present: for centers 2, 3, and 4, the pneumonitis incidence was about 13%, whereas for center 5 only 3%. Explicitly accounting for these differences by adding center dependent offset values for the Lung group, improved the data fit significantly (p < 10(-5)) with NTD50=30.5+/-1.4 Gy and m=0.30+/-0.02 (+/-1 SE) for all patients, and an offset of 0-11% for the Lung group, depending on the center. CONCLUSIONS: The mean lung dose, NTDmean, is relatively easy to calculate, and is a useful predictor of the risk of radiation pneumonitis. The observed dose-effect relation between the NTDmean and the incidence of radiation pneumonitis, based on a large clinical data set, might be of value in dose-escalating studies for lung cancer. The validity of the obtained dose-effect relation will have to be tested in future studies, regarding the influence of confounding factors and dose distributions different from the ones in this study. PMID- 9747814 TI - Lung and heart dose volume analyses with CT simulator in radiation treatment of breast cancer. AB - PURPOSE: Radiation pneumonitis and cardiac effects are directly related to the irradiated lung and heart volumes in the treatment fields. The central lung distance (CLD) from a tangential breast radiograph is shown to be a significant indicator of ipsilateral irradiated lung volume. Retrospective analysis of the pattern of dose volume of lung and heart with actual volume data from a CT simulator in the treatment of breast cancer is presented with respect to CLD. METHODS AND MATERIALS: The heart and lung volumes in the tangential treatment fields were analyzed in 108 consecutive cases (52 left and 56 right breast) referred for CT simulation. All patients in this study were immobilized and placed on an inclined breast board in actual treatment setup. Both arms were stretched over head to avoid collision with the scanner aperture. Radiopaque marks were placed on the medial and lateral borders of the tangential fields. All patients were scanned in spiral mode with slice width and thickness of 3 mm each, respectively. The lung and heart structures as well as irradiated areas were delineated on each slice and respective volumes were accurately measured. The treatment beam parameters were recorded and the digitally reconstructed radiographs (DRRs) were generated for the measurement of the CLD and analysis. RESULTS: Using CT data the mean volume and standard deviation of left and right lungs were 1307.7+/-297.7 cm3 and 1529.6+/-298.5 cm3, respectively. The magnitude of irradiated volume in left and right lung is nearly equal for the same CLD that produces different percent irradiated volumes (PIV). The left and right PIV lungs are 8.3+/-4.7% and 6.6+/-3.7%, respectively. The PIV data have shown to correlate with CLD with second- and third-degree polynomials; however, in this study a simple straight line regression is used to provide better confidence than the higher order polynomials. The regression lines for the left and right breasts are very different based on actual CT data. The slopes of regression lines for the left and right lung are 0.6%/mm and 0.5%/mm, respectively which is statistically different with thep value of 0.01. A maximum heart PIV of >3.0% is observed in 80% of the patients. The heart PIV is inversely correlated with gantry angle and weakly correlated with CLD. CONCLUSIONS: The CT-simulator provides accurate volumetric information of the heart and lungs in the treatment fields. The lung PIV is directly correlated to the CLD (0.6%/mm and 0.5%/mm for the left and right lungs). Left and right lungs have different volumes and hence, different regression lines are recommended. An additional 12% lung volume could be irradiated in the supraclavicular field. Heart volume is not correlated with the CLD. The heart PIV is associated to the beam angle. Heart volume may not be accurately visualized in a tangential radiograph; however, this can be easily seen in a DRR with contour delineation and can be minimized with proper beam parameters iteratively with a virtual simulator. Lung and heart PIV along with dose volume histograms (DVH) are essential in reducing pulmonary and cardiac complications. PMID- 9747815 TI - Predictive factors for late toxicity after endobronchial brachytherapy: a multivariate analysis. AB - PURPOSE: To determine the predictive factors associated with hemoptysis and radiation bronchitis after endobronchial brachytherapy by univariate and multivariate analyses METHODS AND MATERIALS: One hundred forty-nine patients underwent endobronchial brachytherapy and were divided into three therapeutic groups: group 1: patients treated with palliative intent (n=47); group 2: patients treated with curative intent (small endobronchial tumors without mediastinal or general dissemination: n=73); group 3: patients also receiving external irradiation (n=29). One hundred twelve patients had previously received external irradiation. Brachytherapy was delivered with a dose per fraction ranging from 4 to 7 Gy and a prescription point between 0.5 and 1.5 cm, usually 1 cm from the source center. Two to six fractions were delivered according to the therapeutic group and clinical situation. The influence of the following variables on the incidence of hemoptysis or radiation bronchitis was studied: age, sex, Karnofsky score, therapeutic group, histologic type, endoscopic tumor length, dose per fraction, total brachytherapy dose, total external beam irradiation dose, total dose (brachytherapy dose plus external irradiation dose), volumes of the 100% and 200% isodoses, and volumes of the 7 and 14 Gy isodoses. RESULTS: We observed 11 hemoptyses (7.4%), 10 were lethal. All but one occurred in patients with progressive disease. Two clinical factors were significantly associated with hemoptysis by univariate analysis: palliative group (p=0.009) and endobronchial tumor length (p=0.004). No technical factors seem to be implicated in the occurrence of hemoptysis. Only endobronchial tumor length remained in the multivariate model (p=0.02). Radiation bronchitis was observed in 13 cases (8.7%). By univariate analysis, a good Karnofsky score (p=0.02), curative treatment (p=0.02), and tumor location on trachea and main stem bronchus (p=0.002) were significantly associated with this complication. Two technical factors were also incriminated: the total dose (p=0.04) and the 100% isodose volume (p=0.02). By multivariate analysis, only the tumor location retained statistical significance (p=0.009). CONCLUSION: Hemoptysis is most likely due to disease progression, with the bleeding being facilitated by brachytherapy. Some rare cases could be a direct complication of brachytherapy itself, particularly when tumors are located in the upper lobes. In contrast, radiation bronchitis occurred more frequently in patients with controlled disease, and was significantly influenced by tumor location and technical factors (dose and volumes treated). Technical improvements should increase the therapeutic ratio. PMID- 9747817 TI - Oxygenation of squamous cell carcinoma of the head and neck: comparison of primary tumors, neck node metastases, and normal tissue. AB - BACKGROUND: Most previous oxygenation measurements of head and neck tumors have mainly been performed in neck nodes. We investigated, therefore, the relationship between the pO2 status of primary tumors, cervical neck node metastases and normal tissues. PATIENTS AND METHODS: 30 patients with histologically proven advanced stage III-IV squamous cell carcinoma of head and neck underwent pretreatment polarographic pO2 measurements with a pO2 histograph (Eppendorf, Hamburg, Germany). We obtained data on oxygenation of 23 primary tumors, of 22 neck node metastases, and of 30 contralateral sternocleidomastoid muscles. In 15 cases, we were able to perform measurements in all three regions in the same individual. results: A highly significant correlation existed between the median pO2 of primary tumors and their neck node metastases (p=0.0001), as well as between the proportion of pO2 values < or =2.5 mmHg and +/-5.0 mmHg (p=0.0001, p=0.001) in both anatomic sites. The average pretreatment median PO2 was 14.7 mmHg (range 0.2-58.5 mmHg) in primary tumors, 13.7 mmHg (range 1.9-50.3 mmHg) in neck node metastases, and 43.8 mmHg (range 20.8-67.7 mmHg) in sternocleidomastoid muscles. In all cases, the oxygenation of malignant tissue was below that of the corresponding muscle. There was also a weak, but significant, correlation between hemoglobin level and the median pO2 of the primary tumors, as well as between hemoglobin concentration and the proportion of values below 5 mmHg at the primary site (p=0.017, p=0.003). CONCLUSIONS: Primary tumors and their regional lymph node metastases in advanced squamous cell carcinoma of the head and neck show comparable patterns of oxygenation in terms of the median pO2 and the proportion of hypoxic measurements. This report suggests that, in patients with such carcinomas, the oxygenation data obtained at one site are related to tumor oxygenation at other sites, so that measurements in any anatomic site would be sufficient to estimate a tumor's oxygenation status. The weak correlation between pO2 and hemoglobin level requires further investigation. PMID- 9747816 TI - Neoadjuvant chemotherapy and hyperfractionated radiotherapy with concurrent low dose chemotherapy for squamous cell esophageal carcinoma. AB - PURPOSE: We conducted a prospective study of neoadjuvant treatment for squamous cell carcinoma of the esophagus, modifying the chemotherapy protocol by adding l folinic acid and giving bifractionated radiotherapy with a cis diaminedichloroplatinum (CDDP) injection before each fraction. METHODS AND MATERIALS: Thirty-two patients, 30 men, 2 women, mean age 56.2+/-8.9 years, with resectable squamous cell carcinoma of the esophagus (TNM stage I=4, IIA=4, IIB=13, III=11) were included. Chemotherapy, CDDP (80 mg/m2 D2), 5-fluorouracil (5-FU; 600 mg/m2, D1-4), and l-folinic acid (200 mg/m2, D1-4), was given in two sessions with a 3-week interval during which the patients received radiotherapy (45 Gy), two fractions per day (150 cGy/fraction). A 3-mg injection of CDDP was given prior to each fraction. Patients underwent surgery 4 to 7 weeks after neoadjuvant therapy. RESULTS: No severe side effects were observed in 12 patients. Grade 3 effects (WBC, platelets, mucositis) occurred in 16 patients and grade 4 effects (platelets, mucositis) in four including 1 death due to septicemia with an infected catheter. Surgery was performed in 29 patients; 26 had resectable tumors (81%). Operative mortality was 10%. The 26 surgical specimens showed complete response (n=18), persistent microscopic residues (n=4), or not significant modification (n=4). Survival at 1, 2, and 3 years was 81, 61, and 51.6% and disease-free survival was 75, 59, and 54% respectively. CONCLUSIONS: This new therapeutic combination is aggressive and associated with a high postoperative mortality but has a remarkable histological effect since complete response was achieved in 56% (95% CI: 39-73%) of the patients and 3-year survival reached 52%, a very high rate in our experience. PMID- 9747818 TI - Phase I dose escalating trial of hyperfractionated pre-operative chemoradiation for locally advanced rectal cancer. AB - PURPOSE: To determine the acute toxicity, post-operative complications, pathologic response and extent of downstaging to high dose pre-operative radiation using a hyperfractionated radiation boost and concurrent chemotherapy in a prospective Phase I trial. MATERIALS & METHODS: To be eligible for this study, patients had to have adenocarcinoma of the rectum less than 12 cm from the anal verge with either Stage T4 or T3 but greater than 4 cm or greater than 40% of the bowel circumference. All patients received 45 Gy pelvic radiation (1.8 Gy per fraction). Subsequent radiation was given to the region of the gross tumor with a 2 cm margin. This "boost" treatment was given at 1.2 Gy twice daily to a total dose of 54.6 Gy for Level I, 57 Gy for Level II, and 61.8 Gy for Level III. 5-FU was given at 1g/m2 over 24 hours for a four day infusion during the first and sixth weeks of radiation, with the second course concurrent with the hyperfractionated radiation. Surgical resection was carried out 4-6 weeks following completion of chemoradiation (in curative cases) and additional adjuvant chemotherapy consisting of 5-FU and Leucovorin was given for an additional 4 monthly cycles Days 1 through 5 beginning four weeks post surgery. RESULTS: Twenty-seven patients, age 40-82 (median 61), completed the initial course of chemoradiation and are included in the analysis of toxicity. The median follow-up is 27 months (range 8-68). Eleven patients were treated to a dose of 54.6 Gy, nine patients to 57 Gy, and seven patients to 61.8 Gy. Twenty-one patients had T3 tumors, and six patients T4 tumors. Grade III acute toxicity from chemoradiation included proctitis (5 patients), dermatitis (9), diarrhea (five), leukopenia (1), cardiac (1). Grade IV toxicities included one patient with diarrhea (on dose Level I) and one patient (on dose Level III) with cardiac toxicity (unrelated to radiation). Surgical resection consisted of abdominal perineal resection in 16 and low anterior resection in 7. Four patients did not undergo a curative resection; three initially presented with metastases and one developed metastasis during the pre-operative regimen. Post-operative complications included pelvic or perineal abscess in two (on dose Levels I & II), and delayed wound healing in two (one of whom, on dose Level III, developed perineal wound dehiscence requiring surgical reconstruction). Of the 23 patients who had a curative resection, four manifested pathologic complete responses (17.4%). Thirteen of 23 patients (57%) had evidence of pathologic downstaging and only 1/23 patients (on dose Level I) had a positive resection margin. Of these 23 patients (with a minimum follow-up of 8 months), the patient with positive margins was the only one who developed a local failure (Fisher's Exact p=.04). The 3-year actuarial OS, DFS and LC rates are 82%, 72% and 96%, respectively. Twelve of 13 patients (92% at 3 years) > or = 61 years vs. 5/10 patients (45% at 3 years) < 61 years remained disease-free (log-rank p=0.017). CONCLUSION: This regimen of high dose pre-operative chemoradiation employing a hyperfractionated radiation boost is feasible and tolerable and results in significant downstaging in locally advanced rectal cancer. The vast majority of patients (96%) achieved negative margins, which appears to be a prerequisite for local control (p= 0.04). Older age (> or =61 years) was a significant predictor for improved DFS. This regimen (at dose Level III, 61.8 Gy) is currently being tested in a Phase II setting. PMID- 9747819 TI - Sphincter preservation in rectal cancer with preoperative radiation therapy and coloanal anastomosis: long term follow-up. AB - BACKGROUND: To determine if preoperative radiation therapy allows sphincter preservation in the treatment of rectal cancer. METHODS: Thirty six patients with the diagnosis of invasive, resectable, primary adenocarcinoma of the rectum limited to the pelvis were enrolled on a Phase I/II trial of preoperative radiation therapy plus low anterior resection/coloanal anastomosis. By preoperative assessment, all patients had invasive tumors (5,T2; 31,T3) involving the distal half of the rectum and clinically required an abdominoperineal resection. The median tumor size was 3.8 cm [range: 1.5-7 cm] and the median distance from the anal verge was 4 cm [range: 3-7 cm]. The whole pelvis received 46.80 Gy followed by a 3.60 Gy boost to the primary tumor bed. The median follow up was 56 months [range: 4-121 months]. RESULTS: Of the 35 patients who underwent resection, 5 (14%) had a complete pathologic response and 27 (77%) were able to successfully undergo a low anterior resection/coloanal anastomosis. The incidence of local failure was crude: 17% and 5-year actuarial: 21%. The 5-year actuarial survival was 64%. Analysis of sphincter function using a previously published scale was performed at the time of last follow-up in the 27 patients who underwent a low anterior resection/coloanal anastomosis. Function was good or excellent in 85%. The median number of bowel movements/day was 2 (range: 0-8). CONCLUSIONS: Our data suggest that preoperative radiation therapy allows sphincter preservation in 77% of selected patients who would otherwise require an abdominoperineal resection, and 85% have good to excellent sphincter function. Given the moderate local failure rate, we now routinely use preoperative combined modality therapy plus postoperative chemotherapy for patients with clinical T3 disease. PMID- 9747820 TI - Adjuvant therapy for pancreatic cancer: back to the future. AB - PURPOSE: The role of adjuvant therapy in the management of pancreatic cancer, resected with curative intent, remains controversial. This editorial review updates the status of adjuvant therapy in this context and introduces the first North American co-operative group study in this arena in roughly 20 years. RESULTS: To the extent that there has been a "standard" of care in this context, it has been defined in large part by the early work of the Gastrointestinal Study Group (GITSG). Their trial was activated in the mid 1970's using split course radiation therapy and bolus 5-FU. In the intervening 20 + years the morbidity/mortality of pancreaticoduodenectomy (PDD) has been dramatically reduced; concurrently, understanding of prognostic factors impacting on outcomes for resected patients has been significantly enhanced. In major centers the mortality of PDD is roughly 1% and survival has been shown to correlate with a number of factors including tumor size, nodal involvement, and margin status. With currently available techniques doses of continuous course radiation therapy in the range of 50-55 Gy to sites of pancreatic tumor resection and adjacent lymph node regions have been given in a number of trials with acceptable morbidity. 5-FU sequencing and administration have been advanced and gemcitabine, an agent with clear radiosensitizing properties, has been approved for use against pancreatic cancer. CONCLUSIONS: Following PDD increasing numbers of physiologically intact patients are confronting the survival statistics associated with resected pancreatic cancer. Their interest in improved therapeutic outcomes, combined with the noted improvements in radiation and chemotherapeutic management, has set the stage for renewed and intensified study. Accordingly, the intergroup mechanism of the Cancer Therapy and Evaluation Program (CTEP) of the NCI has designed, approved, and activated a modern Phase III, adjuvant protocol incorporating recently gained knowledge in this management context. Prospective randomization will be utilized to compare gemcitabine and 5 FU as single agents before and after chemoradiotherapy with 5-FU. Successful and timely completion of this newly activated intergroup study, RTOG 97-04, will establish a current, cooperative group experience, data base, and standard in the context of adjuvant therapy for pancreatic cancer and serve to provide momentum for further studies. PMID- 9747821 TI - Cost- and time-sparing simplified conformal therapy for prostate cancer: is it feasible? AB - PURPOSE: It was hypothesized that using a simplified technique of volumes profiles determination (STVPD) based on CT data sets (correlate and projection) would increase the target dose without increasing the bladder and rectal dose obtained by conventional simulation techniques. To test this hypothesis, patients referred for radical radiation treatment for prostate carcinoma were prospectively evaluated by performing treatment planning using standard simulation, with (SSB) and without corner blocks (SSWB), STVPD, and 3D beam's eye view. METHODS AND MATERIALS: Twenty-one patients with prostate carcinoma (stage B: 7; stage C: 14) underwent four treatment planning procedures where the field arrangement was defined by standard simulation (SSB and SSWB), STVPD, and 3D beam's eye view (BEV) with a four field (10 MV photons) box technique. Dose volume histograms (DVHs) for the planning target volume (PTV), bladder, and rectum (relatives to the four techniques) were generated for all patients and compared; average percentage dose to the bladder and rectum were also calculated. RESULTS: STVPD and 3D BEV treated an increased percentage of PTV at 95% isodose level, in comparison to standard simulation (with and without blocks). No statistically significant differences were found between the two techniques. A significant reduction of irradiated bladder volume was found between 3D BEV and STVPD versus simulation with and without blocks (mean percentage dose: 77.3%, 81.8%, 93.5%, and 92.6% respectively). No marked differences were recorded in rectal irradiation (mean percentage dose: 53.1%, 53.7%, 51.9%, and 50.2% respectively). Time required for treatment planning (excluding CT scan and definitive simulation) was less than 15 minutes for STVPD and more than 120 minutes for 3D BEV. CONCLUSION: Our results confirm the inadequacy of standard simulation. It is possible, with conformal therapy, to increase the dose to the PTV, decreasing the irradiated volume of the bladder. The absence of sparing effect in the rectum is discussed. Using a box technique, STVPD can be used routinely to define the PTV in patients with prostate cancer, reducing the time required for treatment planning, with dosimetric results similar to those of 3D BEV. PMID- 9747822 TI - Differences in gross target volumes on contrast vs. noncontrast CT scans utilized for conformal radiation therapy treatment planning for prostate carcinoma. AB - PURPOSE: To compare the gross target volumes (GTVs) (prostate and seminal vesicles) defined on noncontrast and contrast-enhanced computed tomography (CT) images used for three-dimensional conformal treatment planning (3DCRT). METHODS AND MATERIALS: From 1993 to 1996, 39 patients referred for radiation therapy for adenocarcinoma of the prostate underwent pretreatment pelvic CT scanning with and without intravenous (i.v.) contrast for treatment planning purposes. Seven patients were excluded because of incomplete data sets. The prostate and seminal vesicles were outlined by the same physician on all images of 32 patients. On 18 CT exams, the prostate and seminal vesicles were blindly outlined a second time by the same physician to evaluate intraphysician consistency. Discrepancies between the GTVs outlined with and without contrast and between the first and second outline on the same study were assessed by calculating the projected area in the anterior-to-posterior (AP) and right lateral (RLAT) beam's-eye view (BEV). To assess the magnitude, frequency, and direction of discrepancies between the two GTVs, the extension of the GTVs in six directions (right, left, anterior, posterior, cephalad, and caudal) was determined. RESULTS: The GTV outlined with contrast was larger in all directions, except caudal, in the majority of patients. The change in the GTV with contrast was significant in the cephalad (p=0.0003) and right (p=0.0007) directions, but not in the other directions. Although the increase with contrast in any direction was usually small (average < or =5 mm), these changes resulted in a significant increase in GTV area in both the AP and RLAT BEV (9.0%, p=0.0017 and 8.2%, p=0.023, respectively). The intraphysician variability in outlining the prostate/ seminal vesicles was minimal. CONCLUSIONS: The addition of i.v. contrast does appear to make a significant difference in how the prostate and seminal vesicles are outlined by an experienced observer. The increase in area of the target, found when contrast is used, should be taken into consideration when designing the treatment fields for patients with carcinoma of the prostate. PMID- 9747824 TI - Cost minimization analysis of high-dose-rate versus low-dose-rate brachytherapy in endometrial cancer. Gynecology Tumor Group. AB - PURPOSE: Endometrial cancer is a common, usually curable malignancy whose treatment frequently involves low-dose-rate (LDR) or high-dose-rate (HDR) brachytherapy. These treatments involve substantial resource commitments and this is increasingly important. This paper presents a cost minimization analysis of HDR versus LDR brachytherapy in the treatment of endometrial cancer. METHODS AND MATERIALS: The perspective of the analysis is that of the payor, in this case the Ministry of Health. One course of LDR treatment is compared to two courses of HDR treatment. The two alternatives are considered to be comparable with respect to local control, survival, and toxicities. Labor, overhead, and capital costs are accounted for and carefully measured. A 5% inflation rate is used where applicable. A univariate sensitivity analysis is performed. RESULTS: The HDR regime is 22% less expensive compared to the LDR regime. This is $991.66 per patient or, based on the current workload of this department (30 patients per year) over the useful lifetime of the after loader, $297,498 over 10 years in 1997 dollars. CONCLUSION: HDR brachytherapy minimizes costs in the treatment of endometrial cancer relative to LDR brachytherapy. These results may be used by other centers to make rational decisions regarding brachytherapy equipment replacement or acquisition. PMID- 9747823 TI - Preoperative chemoradiation for advanced vulvar cancer: a phase II study of the Gynecologic Oncology Group. AB - PURPOSE: To determine the feasibility of using preoperative chemoradiotherapy to avert the need for more radical surgery for patients with T3 primary tumors, or the need for pelvic exenteration for patients with T4 primary tumors, not amenable to resection by standard radical vulvectomy. METHODS AND MATERIALS: Seventy-three evaluable patients with clinical Stage III-IV squamous cell vulvar carcinoma were enrolled in this prospective, multi-institutional trial. Treatment consisted of a planned split course of concurrent cisplatin/5-fluorouracil and radiation therapy followed by surgical excision of the residual primary tumor plus bilateral inguinal-femoral lymph node dissection. Radiation therapy was delivered to the primary tumor volume via anterior-posterior-posterior-anterior (AP-PA) fields in 170-cGy fractions to a dose of 4760 cGy. Patients with inoperable groin nodes received chemoradiation to the primary vulvar tumor, inguinal-femoral and lower pelvic lymph nodes. RESULTS: Seven patients did not undergo a post-treatment surgical procedure: deteriorating medical condition (2 patients); other medical condition (1 patient); unresectable residual tumor (2 patients); patient refusal (2 patients). Following chemoradiotherapy, 33/71 (46.5%) patients had no visible vulvar cancer at the time of planned surgery and 38/71 (53.5%) had gross residual cancer at the time of operation. Five of the latter 38 patients had positive resection margins and underwent: further radiation therapy to the vulva (3 patients); wide local excision and vaginectomy necessitating colostomy (1 patient); no further therapy (1 patient). Using this strategy of preoperative, split-course, twice-daily radiation combined with cisplatin plus 5-fluorouracil chemotherapy, only 2/71 (2.8%) had residual unresectable disease. In only three patients was it not possible to preserve urinary and/or gastrointestinal continence. Toxicity was acceptable, with acute cutaneous reactions to chemoradiotherapy and surgical wound complications being the most common adverse effects. CONCLUSION: Preoperative chemoradiotherapy in advanced squamous cell carcinoma of the vulva is feasible, and may reduce the need for more radical surgery including primary pelvic exenteration. PMID- 9747825 TI - Carcinoma of the uterine cervix in Saudi Arabia: experience in the management of 164 patients with stage-I & -II disease. AB - BACKGROUND: Earlier stages cervical cancer has been customarily treated with radiation therapy, surgery, or combination thereof. We present our experience in the management of stage-I and -II patients in a major cancer center in the Kingdom of Saudi Arabia. METHODS: Between 1979 and 1991, 164 patients were treated and closely followed at a tertiary care medical center. RESULTS: Patients accounted for 0.78 % of all cancer patient referral. More cases with earlier stages (41.3 %) were referred since 1986, compared to a lower referral (26.1%) during the earlier part of the study (p=0.027). Age ranged from 21 to 80 years with a median of 46.5 years. Clinical stages at presentation included Stage-IA (3.0%), IB (28.7%), IIA (11.6%), and IIB (56.7%). Majority (87.2%) had squamous cell carcinoma, while the rest, had adenocarcinoma (9.1%) or other malignancies (3.6%). Among the 143 patients with squamous cell cancer, eighteen had attempted radical resection, 101 were treated with radiation, and 24 had both modalities. For squamous cancer patients, fifty-one (35.7%) had disease relapse either locally (19 cases), distally (23 patients) or both combined (9 patients). The pattern of failure was unrelated to stage of disease, histological diagnosis or the mode of therapy initially administered. The cumulative five and ten year's survival for squamous cancer patients was 68.3% and 57.9% respectively. Better survival was noted for patients with smaller sized tumors, free parametrium, and Stage-I disease. When all factors were considered in the regression model, only the status of parametrial involvement was found to be of significance. CONCLUSIONS: Cervical cancer is relatively rare in Saudi Arabia. With the improvement in health care delivery, more patients were lately seen at earlier stages of disease. With radiation therapy, two thirds of patients survived five years. The extent of parametrial involvement was the best predictor for long term survival. PMID- 9747826 TI - Is there a role for a brachytherapy vaginal cuff boost in the adjuvant management of patients with uterine-confined endometrial cancer? AB - PURPOSE/OBJECTIVE: Many patients who have uterine-confined endometrial cancer with prognostic factors predictive of recurrence are treated with adjuvant pelvic radiation. The addition of a brachytherapy vaginal cuff boost is controversial. MATERIALS AND METHODS: Between 1983 and 1993, 270 patients received adjuvant postoperative pelvic irradiation following hysterectomy for Stage I or II endometrial cancer. Group A includes 173 patients who received external beam irradiation alone (EBRT), while group B includes 97 patients who received EBRT with a vaginal brachytherapy application. The median dose of EBRT was 45 Gy. Vaginal brachytherapy consisted of a low dose rate ovoid or cylinder in 41 patients, a high dose rate cylinder in 54 patients, and a radioactive gold seed implant in two patients. The median follow-up time was 64 months. The two groups were compared in terms of age, histologic grade, favorable versus unfavorable histology, capillary space invasion, depth of myometrial invasion, and pathologic stage. RESULTS: Chi-square analysis revealed that the only difference between the two groups was the presence of more Stage II patients in group B (38% versus 14%). No difference was detected for 5 year pelvic control and disease-free survival rates between groups A and B. CONCLUSION: There is no suggestion that the addition of a vaginal cuff brachytherapy boost to pelvic radiation is beneficial for pelvic control or disease-free survival for patients with Stage I or II endometrial cancer. Prospective randomized trials designed to study external irradiation alone versus external beam treatment plus vaginal brachytherapy are unlikely to show a positive result. Because EBRT provides excellent pelvic control, protocol development for uterine-confined corpus cancer should focus on identifying patients at risk for recurrence as well as other means of augmenting EBRT (e.g. addition of chemotherapy) in order to improve disease free survival in those subgroups. PMID- 9747827 TI - The presence of proliferative breast disease with atypia does not significantly influence outcome in early-stage invasive breast cancer treated with conservative surgery and radiation. AB - PURPOSE: To evaluate the influence of the benign background breast-tissue change of atypical hyperplasia (AH) on outcome in patients with early-stage invasive breast cancer treated with conservative surgery and radiation. MATERIALS AND METHODS: Four hundred and sixty women with Stage I--II breast cancer treated with conservative surgery and radiation from 1982-1994 had pathologic assessment of their background adjacent benign breast tissue. The median follow-up was 5.6 years (range 0.1-15). The median age was 55 years (range 24-88). Of these, 23% had positive axillary nodes; 25% received adjuvant chemotherapy (CMF or CAF) with (9%) or without (17%) tamoxifen. Of the total, 24% received adjuvant tamoxifen alone. The patients were divided into 2 groups: 131 patients with atypical hyperplasia (ductal, 99 patients; lobular, 20 pts; and type not specified, 12 pts), and 329 patients with no proliferative changes or proliferative changes without atypia. RESULT: A statistically significant difference was observed between the 2 groups for method of detection, primary tumor size, presence of lobular carcinoma in situ (LCIS), pathologic nodal status, region(s) treated with radiation, and type of adjuvant therapy. Patients with atypical hyperplasia (AH) had smaller primary tumors (T1 80% vs. 70%) more often detected solely by mammography (51% vs. 36%) with negative axillary nodes (87% vs. 73%) and radiation treatment to the breast only (93% vs. 78%). LCIS was observed in 9% of the patients with AH and 3% of those without AH. Patients with AH more often received tamoxifen alone (32% vs. 21%), rather than chemotherapy (15% vs. 29%). There were no statistically significant differences between the 2 groups for race, age, menopausal status, family history, histology, histologic subtype DCIS when present, the presence or absence of an extensive intraductal component, final margin status, estrogen or progesterone receptor status, use of re excision, or total radiation dose to the primary. The 5- and 10-year actuarial ipsilateral breast tumor recurrence rates were 2% and 12% for patients with AH and 4% and 8% for those without AH (p=0.44). Younger women or those with a positive family history of breast cancer with AH did not have an increased rate of breast failure when compared to similar patients without AH. There were no significant differences in the 5- and 10-year actuarial rates of distant metastases (AH 5- and 10-year 7% and 7%, no AH 5- and 10-year 8% and 16%,p=0.31), regional node recurrence (AH 1% and 1%, no AH 1% and 1%,p=0.71), contralateral breast cancer (AH 3% and 3%, no AH 3% and 8%,p=0.71), overall survival (AH 95% and 86%, no AH 95% and 89%, p=0.79), or cause-specific survival (AH 98% and 95%, no AH 96% and 91%,p=0.27). Subset analysis for ipsilateral breast tumor recurrence, distant metastases, overall, and cause-specific survival for T1 vs. T2 tumors and path node-negative vs. path node-positive patients revealed no significant differences between the 2 groups. CONCLUSION: AH was not associated with an increased risk of ipsilateral breast tumor recurrence or contralateral breast cancer in this study of patients with invasive breast cancer treated with conservative surgery and radiation. Therefore, the presence of proliferative changes with atypia in background benign breast tissue should not be a contraindication to breast-conservation therapy. PMID- 9747828 TI - Long-term follow-up of axillary node-positive breast cancer patients receiving adjuvant tamoxifen alone: patterns of recurrence. AB - PURPOSE: To determine the patterns, incidence and risk factors for local-regional recurrence in patients with Stage II and III breast cancer treated with adjuvant tamoxifen alone, without adjuvant radiation. MATERIAL AND METHODS: The records of patients referred to the London Regional Cancer Centre with a diagnosis of breast cancer between 1980-1989 were reviewed. During this time period, it was the policy of the institution to omit local-regional radiation to patients receiving adjuvant systemic therapy. One hundred and fifty axillary node-positive Stage II and III breast cancer patients received adjuvant tamoxifen alone without postoperative local-regional radiation; these patients form the basis of this report. RESULTS: Median follow-up was 67 months for the entire patient group and 85 months for the living patients. During this time, 42% of patients developed a recurrence, 22% first recurred in local-regional sites. The total incidence of local-regional recurrence (including those patients who first relapsed with systemic metastases) was 30%. Of the segmental mastectomy patients, 13% had recurrences in the intact breast. Of the modified radical mastectomy patients, 10% developed chest wall recurrences. Five percent of recurrences were first in the axilla and 6% in the supraclavicular nodes. Five-year actuarial survival for the entire patient group was 79% and disease-free survival was 60%, with a median disease-free survival time of 87 months. Five-year local-regional relapse-free survival was 76%. Five-year local-regional relapse-free survival was < 76% for those patients with 4 or more positive axillary nodes, regardless of tumor size. On univariable analysis, positive resection margins, number of positive axillary nodes, menopausal status, and negative estrogen and progesterone receptors were significant for isolated local-regional recurrence. On multivariable analysis, only positive resection margins and negative receptors remained significant. In terms of regional recurrence specifically, negative estrogen and progesterone receptor status and positive resection margins were, again, prognostically significant. CONCLUSIONS: Postmenopausal women receiving adjuvant tamoxifen who have positive resection margins, > or = 4 positive axillary nodes and/or negative estrogen and progesterone receptors, are at higher risk of local and regional recurrence and should, therefore, receive local-regional radiation. PMID- 9747829 TI - A multidisciplinary study investigating radiotherapy in Ewing's sarcoma: end results of POG #8346. Pediatric Oncology Group. AB - PURPOSE: To determine if involved field radiation (IF) is equivalent to standard whole bone radiation (SF) in local tumor control; to establish patterns of failure following treatment; and to determine response, event-free survival (EFS), and overall survival rates from multidisciplinary therapy in Ewing's sarcoma. METHODS AND MATERIALS: Between 1983 and 1988, 184 children with Ewing's sarcoma were enrolled onto Pediatric Oncology Group 8346 (POG 8346). A total of 178 (97%) met eligibility criteria; 6 had pathology other than Ewing's sarcoma. Induction chemotherapy of cyclophosphamide/doxorubicin (adriamycin )(C/A) x 12 weeks was followed by local treatment either surgery or radiation therapy and C/A, dactinomycin, and vincristine for 50 weeks. Resection was advised for patients with small primary tumors if accomplished without functional loss. Forty patients were randomized to receive SF, whole bone radiation to 39.6 Gy plus a 16.2 Gy boost (total 55.8 Gy) or IF to 55.8 Gy, and the remainder were assigned to IF radiation. RESULTS: Of 178 eligible patients, 141 (79%) had localized disease and 37 (21%) had metastases at presentation. Their 5-year EFS was 51% (SE 5%) and 23% (SE 7%) respectively. The response rate to induction chemotherapy was 88% (28% complete, 60% partial), but after radiotherapy the response rate increased to 98%. Thirty-seven of the localized patients underwent resection, of whom 16 (43%) required postoperative radiotherapy; the 5-year EFS of these surgical patients was 80% (SE 7%). The remaining 104 localized patients were eligible for randomization or assignment to receive radiotherapy; the 5-year EFS of these patients was 41% (SE 5%), with no significant difference in EFS between those randomized to SF vs. IF. Site of primary tumor correlated with 5-year EFS: distal extremity 65% (SE 8%), central 63% (SE 10%), proximal extremity 46% (SE 8%), and pelvic-sacral 24% (SE 10%) (p=0.004). Initial tumor size did not correlate significantly with EFS. Patterns of failure among the 141 localized patients revealed 23% of patients experienced a local failure, while 40% had a systemic failure. The 5-year local control rate for the surgical patients +/- postoperative radiotherapy was 88% (SE 6%), while for the patients undergoing radiotherapy alone it was 65% (SE 7%). There was no difference in local control between those randomized to SF vs. IF. The 5-year local control rate for the patients with pelvic-sacral tumors was 44% (SE 15%), significantly worse than the local control rates for those with central tumors 82% (SE 8%), distal extremity 80% (SE 8%), or proximal extremity 69% (SE 9%) (p=0.023). However, quality of radiotherapy correlated with outcome. Patients who had appropriate radiotherapy had a 5-year local control of 80% (SE 7%), while those with minor deviations had 5-year local control of 48% (SE 14%), and those with major deviations had a local control of only 16% (SE 15%) (p=0.005). The local failure was within an irradiated volume in 62% of patients, outside the irradiated volume in 24% of cases, while the precise location could not be determined in the remaining 14%. CONCLUSIONS: As most failures in Ewing's sarcoma are systemic, improved EFS requires more effective systemic chemotherapy. Adequate IF radiotherapy requires treatment to appropriate volumes as defined by MRI imaging and full radiation doses. Pretreatment review of radiologic images with a musculoskeletal radiologist to determine appropriate tumor volumes, as well as use of conformal radiotherapy techniques are important for improved outcome. PMID- 9747830 TI - A brain tumor dose escalation protocol based on effective dose equivalence to prior experience. AB - PURPOSE: The current study describes the design of a dose escalation protocol for conformal irradiation of primary brain tumors that preserves the safe experience of a previous, sequential dose escalation scheme while enabling the delivery of substantially higher effective doses to a central target volume. METHODS AND MATERIALS: Normalized isoeffective composite dose distributions were formed for 20 patients treated on the original protocol (which specified three progressively smaller planning target volumes [PTVs]) using the linear quadratic model (here corrected to equivalent 2 Gy fractions using alpha/beta=10 Gy). These distributions were investigated and a new protocol was designed to preserve a similar level of efficacy and lack of toxicity for the outer volumes, but allowing a higher dose to the inner PTV. Treatment plans were then investigated to determine if the objectives of the new protocol were achievable. In particular, plans that simultaneously achieved all biological treatment planning objectives (all fields treated each day) were investigated. Finally, the success of the protocol design was demonstrated by analysis of the effective dose distributions of 10 patients treated using the new protocol. RESULTS: The composite normalized isoeffective minimum doses to the outer PTVs (PTV3 and PTV2) in the original protocol were close to 60 Gy and 75 Gy, respectively, and these values are specified as the minimum doses to those volumes for the new protocol. Homogeneity requirements to maintain equivalence for the outer target volume domains are: not more than 25% of [PTV3 exclusive of PTV2] >75 Gy; and not more than 50% of [PTV2 exclusive of PTV1] >85 Gy. Treatment plans using multiple noncoplanar arrangements of beams and static intensity modulation treat all volumes at each session. DVHs of the normalized isoeffective dose distributions reveal the equivalence of the new protocol plans to the sequential plans in the previous protocol as well as the ability to achieve a higher dose of 90 Gy to the isocenter of PTV1 (+/-5% homogeneity required). CONCLUSION: The ability to incorporate past experience through use of the linear quadratic model in the design of a new dose escalation protocol is demonstrated. PMID- 9747831 TI - Patterns of failure following treatment for medulloblastoma: is it necessary to treat the entire posterior fossa? AB - PURPOSE: Craniospinal radiation (CSRT) followed by a boost to the entire posterior fossa (PF) is standard postoperative therapy for patients with medulloblastoma. A large proportion of recurrences after treatment are local, with approximately 50-70% of recurrences occurring in the PF. It is unclear, however, whether these failures are occurring in the original tumor bed or outside the tumor bed, but still within the PF. With improved diagnostic imaging, better definition of tumor volumes, and the use of three-dimensional conformal therapy (3D CRT), we may be able to restrict the boost volume to the tumor bed plus a margin without compromising local control. This retrospective study analyzes the patterns of failure within the PF in a series of patients treated with radiation therapy (RT). METHODS: From July 1986 through February 1996, 114 patients >18 months and <18 years with medulloblastoma were treated at the University of Michigan and Children's Hospital of Philadelphia, with RT following surgical resection. Of 114, 27 (24%) were found to have a recurrence and form the basis for this study. RT consisted of CSRT followed by a boost to the entire posterior fossa. Some patients received adjuvant chemotherapy. Patient's preoperative magnetic resonance imaging (MRI) and/or computerized tomography (CT) studies were used to compare the original tumor volume with the specific region of local relapse. Failure was defined as MRI or CT evidence of recurrence or positive cerebrospinal fluid cytology. Relapse was scored as local, if it was within the original tumor bed, and regional if it was outside of the tumor bed but still within the PF. RESULTS: The median age of the 27 patients who relapsed was 8.6 years. Three patients were <3 years old. Of 27, 21 had disease localized to the PF. Of 26, 22 patients received chemotherapy during their treatment regimen; 1 patient did not have information on systemic treatment. The median dose of RT to the craniospinal axis was 32.5 Gy and to the PF was 55.2 Gy. The median time to recurrence was 19.5 months. Local failure within the tumor bed as any component of first failure occurred in 52% (14 of 27) of all failures, but as the solitary site of first failure in only 2 of 27 failures. Of 14 patients who failed in the tumor bed, 11 also failed in the spine, 8 of 14 also failed within the PF but outside the tumor bed, and 7 of 14 failed in all three locations. Local failure within the PF but outside the tumor bed as any component of first failure occurred in 41% (11 of 27) of all failures, but as the solitary site of first failure in only 1 of 27 failures. Of 11 patients who failed in the PF but outside the tumor bed, 9 also failed in the spine, 8 also failed within the tumor bed, and 7 failed in the all three locations. Of the failures outside the tumor bed but still within the PF, 7 of 11 failed in the leptomeninges, 1 in the brainstem parenchyma, and 3 in the PF parenchyma. Of 7 who failed in the PF leptomeninges, 6 also failed within the spine. Failure within the spine as any component of first failure occurred in 70% (19 of 27) of all failures and as the only site of first failure in 5 of 27 patients. Of 19 patients who failed in the spine, 11 also failed in the tumor bed, 9 also failed within the PF but outside the tumor bed, and 9 failed in the all three locations. CONCLUSIONS: Leptomeningeal failure is a common component of failure and occurs in the leptomeninges of the PF, as well as the spine. Isolated tumor bed failure is a rarely observed event and occurred in only 2 of 27 failures described here. Similarly, parenchymal (nonleptomeningeal) failures in the PF but outside of the tumor bed were rare: 4 patients recurred in this manner, only 1 of whom was an isolated event without other sites of recurrence. Our data suggest that, when the entire PF is treated, very few failures develop in isolation in the PF outside the tumor bed. Further studies will be necessary to determine if RT to the tu PMID- 9747832 TI - Medulloblastoma: time-dose relationship based on a 30-year review. AB - PURPOSE: Time-dose relationships have proven important in many cancer sites. This study evaluates the time factors involved in the successful postoperative radiotherapy of medulloblastoma, based on a 30-year experience in a single institution. METHODS AND MATERIALS: Fifty-three patients with medulloblastoma received postoperative craniospinal radiotherapy with curative intent between 1963 and 1993. Seven patients (13%) underwent biopsy alone, 28 patients (53%) had subtotal excision, and 18 patients (34%) had gross total excision. Eleven patients received adjuvant chemotherapy. The mean posterior fossa dose was 53.1 Gy; most patients received 54.0 Gy (range, 34.3 to 69.6 Gy). For 41 patients receiving once-a-day therapy, the mean dose was 50.6 Gy (range, 34.3 to 56.0 Gy). For 12 patients receiving twice-a-day therapy, the mean dose was 61.8 Gy (range, 52.6 to 69.6 Gy). Minimum follow-up was 2 years, and median follow-up was 10.7 years. Survival, freedom from relapse, and disease control in the posterior fossa were calculated using the Kaplan-Meier method, and multivariate analysis was performed for prognostic factors. Variables related to radiotherapy were examined, including dose to the craniospinal axis, dose to the posterior fossa, fractionation (once-a-day vs. twice-a-day), use of adjuvant chemotherapy, risk group [high (> or =T3b or > or =M1) or low (< or =T3a and M0-MX)], interval between surgery and radiotherapy (excluding patients receiving chemotherapy before radiotherapy), and duration of radiotherapy. RESULTS: At 5 and 10 years, overall survival rates were 68 and 64%, respectively, and freedom-from-relapse rates were 61 and 52%, respectively. Rates of disease control in the posterior fossa at 5 and 10 years were 79 and 68%, respectively. At 5 years, absolute survival rates after biopsy alone, subtotal excision, and gross total excision were 43, 67, and 78%, respectively (p=0.04), and posterior fossa control rates were 27, 89, and 83%, respectively (p=0.004). Duration of the treatment course was the only radiotherapy-related variable with a significant impact on freedom from relapse and posterior fossa control. For patients whose radiation treatment duration was < or =45 days, posterior fossa control was 89% at 5 years, compared with 68% for those treated for >45 days (p=0.01). Duration of treatment also affected freedom from relapse at 5 years: < or =45 days (76%) compared with >45 days (43%), p=0.004. CONCLUSION: Our study demonstrates that if adequate doses are used, then radiotherapy treatment duration will significantly affect the outcome in terms of control of disease in the posterior fossa and freedom from relapse. Fractions of at least 1.75 Gy given once a day, or a twice-a-day regimen should yield optimal local control results. PMID- 9747833 TI - Prognostic factors derived from recursive partition analysis (RPA) of Radiation Therapy Oncology Group (RTOG) brain metastases trials applied to surgically resected and irradiated brain metastatic cases. AB - PURPOSE: (a) To identify the prognostic factors that determine survival after surgical resection and irradiation of tumors metastatic to brain. (b) To determine if the prognostic factors used in the recursive partition analysis (RPA) of brain metastases cases from Radiation Therapy Oncology Group (RTOG) studies into three distinct survival classes is applicable to surgically resected and irradiated patients. METHOD: The medical records of 125 patients who had surgical resection and radiotherapy for brain metastases from 1985 to 1997 were reviewed. The patients' disease and treatment related factors were analyzed to identify factors that independently determine survival after diagnosis of brain metastasis. The patients were also grouped into three classes using the RPA derived prognostic parameters which are: age, performance status, state of the primary disease, and presence or absence of extracranial metastases. Class 1: patients < or = 65 years of age, Karnofsky performance status (KPS) of > or =70, with controlled primary disease and no extracranial metastases; Class 3: patients with KPS < 70. Patients who do not qualify for Class 1 or 3 are grouped as Class 2. The survival of these patients was determined from the time of diagnosis of brain metastases to the time of death. RESULTS: The median survival of the entire group was 9.5 months. The three classes of patients as grouped had median survivals of 14.8, 9.9, and 6.0 months respectively (p=0.0002). Age of < 65 years, KPS of > or = 70, controlled primary disease, absence of extracranial metastases, complete surgical resection of the brain lesion(s) were found to be independent prognostic factors for survival; the total dose of radiation was not. CONCLUSION: Based on the results of this study, the patients and disease characteristics have significant impact on the survival of patients with brain metastases treated with a combination of surgical resection and radiotherapy. These parameters could be used in selecting patients who would benefit most from such treatment. PMID- 9747834 TI - A randomized trial of three single-dose radiation therapy regimens in the treatment of metastatic bone pain. AB - PURPOSE: To investigate efficacy of three single dose radiation therapy (RT) regimens in the treatment of painful bone metastasis. MATERIAL AND METHODS: Patient self-assessment by using pain chart enabled evaluation of response to treatment that consisted of either one of the three single fractions of 4 Gy (group I; n=109), 6 Gy (group II; n=108), or 8 Gy (group III; n=110). RESULTS: Patients in groups II and III had higher complete response rate than those in group I, but not significantly, and with no difference between group II and III. However, both patients in group II (73%) and group III (78%) had significantly higher overall response rates when compared to those observed in group I (59%) (I vs II, p=0.025; I vs III, p=0.0019), and with no difference between groups II and III (p=0.39). Patients in group III had shortest time to the occurrence of any pain relief which was significantly better than those observed in group I (Welch's t-test, p=0.012), with no difference between group I and II and group II and III, respectively. There was no difference between the three treatment groups in duration of response and retreatment rate. No effect of histology or metastatic site treated was found. No pathological fractures or spinal cord compressions were observed during the 8 weeks post-RT. CONCLUSION: Results of this study seem to confirm that 8 Gy could be considered as probably "lowest" optimal single fraction RT in the treatment of painful bone metastasis, although single fraction RT of 4 Gy should not be easily discarded due to its applicability in specific cases. Since single fraction RT of 6 Gy achieved results not different from that obtained with 8 Gy, further studies are warranted in order to get more informations about "lowest" optimal single fraction RT in the treatment of painful bone metastasis. PMID- 9747835 TI - Changes in histology and fibrogenic cytokines in irradiated colorectum of two murine strains. AB - PURPOSE: A strain difference in the development of radiation-induced fibrosis of the colorectum was recently observed. C57B1/6 mice developed colorectal obstruction with significantly higher incidence compared to C3Hf/Kam mice after partial volume irradiation with 30 Gy. Previous reports have demonstrated differences in cytokine mRNA levels in fibrosis-prone and -resistant mice after lung irradiation. The aims of this study are to determine if there are strain differences in: 1) the histology of the lesion, 2) mRNA levels for transforming growth factor beta (TGFbeta) isoforms and tumor necrosis factor alpha (TNFalpha), and 3) immunohistochemical staining patterns using antibodies against the TGFbeta isoforms and latency-associated peptide (LAP). METHODS AND MATERIALS: The colorectum of male C3Hf/Kam (C3H) and C57B1/6 (B6) mice were irradiated using a dose/length combination (30 Gy to 13.7 mm) that resulted in 10 or 100% incidence of obstruction by 6 months in each strain, respectively. Colorectal tissue was removed from 6 hours to 120 days after irradiation as well as from obstructed mice and prepared for histology, RNase protection assay, and immunofluorescence. RESULTS: Distinct differences in the histological phenotype for the two strains were observed at times preceding obstruction. Samples from B6 mice showed increased hyperplastic crypts, colitis cystica profunda, and fibrosis within the lamina propria, compared to identically treated C3H mice. Fibrosis in the lamina propria of B6 mice appeared early, beginning at 75 days after irradiation, and was progressive, whereas fibrosis in C3H mice appeared simultaneous with obstruction and may have been a reaction to ulceration. No consistent strain difference in mRNA levels for TGFbeta1, 2, 3 or TNFalpha were observed, although mRNA levels of TGFbeta1 and TNFalpha were significantly elevated in both strains relative to nonirradiated controls. Immunofluorescent staining for TGFbeta1, 3 and LAP was observed in hyperplastic crypts and colitis cystica profunda adjacent to regions of fibrosis, histological changes that were present predominately in the B6 strain. CONCLUSIONS: The response of the colorectum to irradiation involves changes in the expression of several different cytokines. However, the lack of a consistent strain difference in TGFbeta1, 2, 3 and TNFalpha mRNA levels, despite strain differences in both the incidence of colorectal obstruction and histological features preceding obstruction, suggests that mRNA changes in these fibrogenic cytokines are not the critical determinant of the strain difference and are not related to the process of radiation-induced colorectal fibrosis in these mouse strains. Strain-dependent differences were observed in the localization of active TGFbeta, but these differences were related to the histological changes specifically found in the irradiated colon of the B6 strain. PMID- 9747836 TI - An experimental model of acute encephalopathy after total body irradiation in the rat: effect of liposome-entrapped Cu/Zn superoxide dismutase. AB - PURPOSE: To develop an experimental model of acute encephalopathy following total body irradiation in rats and to define the therapeutic effect of liposome entrapped Cu/Zn superoxide dismutase. METHODS AND MATERIALS: A total of 120 4 month-old rats received 4.5 Gy total body irradiation (TBI) while 120 rats received sham irradiation. A behavioral study based on a conditioning test of negative reinforcement, the one-way avoidance test, was performed 5 hours before irradiation and repeated the following days. Subcutaneous treatment was started 1 hour after irradiation and repeated daily for 2 weeks. In both the irradiated and sham group, three subgroups were defined according to the treatment received: liposome-entrapped Cu/Zn superoxide dismutase (0.5 mg/kg), liposomes only, normal saline. RESULTS: This work comprised two consecutive studies. In study A (90 rats) the one-way avoidance test was administered daily from day 0 to day 4 with a recall session at day 14. In study B (validation phase in 150 rats) the behavioral test was performed only from day 0 to day 6. Before irradiation, all rats showed a similar behavioral response. Study A (6 groups of 15 rats): Following TBI, irradiated rats treated with liposomes only or saline demonstrated a significant delay in learning the one-way avoidance test in comparison with sham-irradiated rats (0.05 < p <0.001 depending upon the day of evaluation and the subgroup type). In contrast, irradiated rats treated with liposome-entrapped Cu/Zn superoxide dismutase did not differ from sham-irradiated rats. Study B (6 groups of 25 rats): The results were the same as those in study A, demonstrating a significant delay in the learning of the test in the liposome and saline treated irradiated rats in comparison with sham-irradiated rats (0.02 < p < 0.001). The irradiated rats, treated with liposome-entrapped Cu/Zn superoxide dismutase did not differ from the sham-irradiated controls. CONCLUSION: This study indicates that a relatively low dose of total body irradiation induces a substantial acute learning dysfunction in the rat. This effect is prevented by the administration of liposome-entrapped Cu/Zn superoxide dismutase. PMID- 9747837 TI - ASTRO research fellowship: the role of BCL-2 and glutathione in an antioxidant pathway to prevent radiation-induced apoptosis. AB - PURPOSE: The expression of the bcl-2 proto-oncogene has been associated with resistance to radiation-induced apoptosis. There is evidence that the bcl-2 protein acts in an antioxidant pathway to block the effects of reactive oxygen species that mediate apoptosis possibly by increasing the levels of intracellular glutathione. Our hypothesis is that pretreatment of radiation-sensitive cells, known to lack bcl-2 expression, with antioxidants will reduce radiation-induced apoptosis. For this purpose, the apoptotic response to radiation and the intracellular levels of GSH were tested before and after pretreatment with antioxidants in two murine lymphoma cell lines, a radiation-resistant, bcl-2- expressing (LY-ar) line and a radiation-sensitive, non-bcl-2-expressing (LY-as) line. METHODS AND MATERIALS: LY-ar and LY-as cells were irradiated at 0,1,2,3, and 4 hours before collection. The intracellular levels of reduced (GSH) and oxidized (GSSG) glutathione were determined by the use of the fluorescent dye o phthalaldehyde. LY-as cells were treated with GSH ethyl-ester for 1 and 2 hours after irradiation. Apoptotic response was measured by the DNA fragmentation assay. The radiation dose was 2.5 Gy. RESULTS: After irradiation, the apoptotic rate of LY-ar and LY-as cells was 10-20% and 50-70% respectively. LY-ar cells had higher intracellular GSH and GSSG levels compared to LY-as cells by 69.9% and 91.9% respectively and the GSH/GSSG ratio in LY-ar and LY-as cells was 15.09 and 17.09 respectively. GSH levels did not change during the first 2 hours after irradiation; however, there was a 49% and 84% reduction at 3 and 4 hours after irradiation, respectively, times at which the LY-as cells have already fragmented their DNA. Treatment of LY-as cells with GSH ethyl-ester at a concentration of 7 mM for 1 and 2 hours resulted in 70% and 231% increases in the intracellular GSH levels respectively. Treatment of LY-as cells with GSH ethyl-ester for 1 and 2 hours also conferred a 25-50% decrease in their apoptotic response after irradiation. CONCLUSIONS: GSH and GSSG levels are elevated in radiation resistant, bcl-2-expressing murine lymphoma cells compared to radiation sensitive, non-bcl-2-expressing cells. GSH levels decline only in radiation sensitive cells after irradiation but this appears to occur at the time of apoptotic cell death. Exogenous thiols increase intracellular GSH levels and repress radiation-induced apoptosis. In conclusion, intracellular thiols appear to be involved in protecting cells from apoptotic cell death. Further investigation should be directed in identifying substances which by lowering intracellular thiols may result in sensitization to radiation. PMID- 9747838 TI - Oxidative damage of mitochondrial and nuclear DNA induced by ionizing radiation in human hepatoblastoma cells. AB - PURPOSE: Since reactive oxygen species (ROS) act as mediators of radiation induced cellular damage, the aim of our studies was to determine the effects of ionizing radiation on the regulation of hepatocellular reduced glutathione (GSH), survival and integrity of nuclear and mitochondrial DNA (mtDNA) in human hepatoblastoma cells (Hep G2) depleted of GSH prior to radiation. METHODS AND MATERIALS: GSH, oxidized glutathione (GSSG), and generation of ROS were determined in irradiated (50-500 cGy) Hep G2 cells. Clonogenic survival, nuclear DNA fragmentation, and integrity of mtDNA were assessed in cells depleted of GSH prior to radiation. RESULTS: Radiation of Hep G2 cells (50-400 cGy) resulted in a dose-dependent generation of ROS, an effect accompanied by a decrease of reduced GSH, ranging from a 15% decrease for 50 cGy to a 25% decrease for 400 cGy and decreased GSH/GSSG from a ratio of 17 to a ratio of 7 for controls and from 16 to 6 for diethyl maleate (DEM)-treated cells. Depletion of GSH prior to radiation accentuated the increase of ROS by 40-50%. The depletion of GSH by radiation was apparent in different subcellular sites, being particularly significant in mitochondria. Furthermore, depletion of nuclear GSH to 50-60% of initial values prior to irradiation (400 cGy) resulted in DNA fragmentation and apoptosis. Consequently, the survival of Hep G2 to radiation was reduced from 25% of cells not depleted of GSH to 10% of GSH-depleted cells. Fitting the survival rate of cells as a function of GSH using a theoretical model confirmed cellular GSH as a key factor in determining intrinsic sensitivity of Hep G2 cells to radiation. mtDNA displayed an increased susceptibility to the radiation-induced loss of integrity compared to nuclear DNA, an effect that was potentiated by GSH depletion in mitochondria (10-15% intact mtDNA in GSH-depleted cells vs. 25-30% of repleted cells). CONCLUSION: GSH plays a critical protective role in maintaining nuclear and mtDNA functional integrity, determining the intrinsic radiosensitivity of Hep G2. Although the DNA repair is a complex process that is not yet completely understood, the protective role of GSH probably does not seem to involve the repair of classical DNA damage but may relate to modification of DNA damage dependent signaling. PMID- 9747839 TI - A solid water pelvic and prostate phantom for imaging, volume rendering, treatment planning, and dosimetry for an RTOG multi-institutional, 3-D dose escalation study. Radiation Therapy Oncology Group. AB - PURPOSE: With increased interest in 3-D conformal radiation therapy and dose escalation, it is necessary to provide advanced techniques to assure quality in treatment delivery. Multi-institutional trials for these newer treatment techniques require methods of verifying the consistency of treatments between the participating institutions. For this reason, a phantom was designed to address the quality and consistency of Radiation Therapy Oncology Group (RTOG) 3-D prostate treatment protocol. METHODS AND MATERIALS: A solid water pelvic and prostate phantom for imaging, volume rendering, treatment planning, and dosimetry applications for performing comprehensive quality assurance has been designed and fabricated. Its configuration was based upon CT slices obtained from a patient study. Individual slices were machined with corresponding contours of the prostate, bladder, rectum, and the left and right femurs. Most of the phantom is made of solid water (Gammex/RMI, Middleton, WI), while the femurs are made of bone-equivalent material. The CT numbers from patient images were used to adjust the solid water composition within the organ volumes, providing image contrast from the remainder of the phantom. Cylindrical insertion grooves are machined in the phantom to allow placement of ionization chambers and thermal luminal dosimeters (TLDs) for dosimetry applications. During imaging, the cavities are filled with rods fabricated from solid water material. RESULTS: The phantom is being used to evaluate the consistency of a range of processes in radiation therapy simulation, planning, and delivery of 3-D-based treatments for prostate cancer. CONCLUSION: The ultimate study objective is to use the phantom to evaluate the accuracy and consistency of treatments delivered by institutions participating in national collaborative clinical trials involving 3-D conformal dose escalation. PMID- 9747840 TI - A critical evaluation of the planning target volume for 3-D conformal radiotherapy of prostate cancer. AB - PURPOSE: To determine an adequate planning target volume (PTV) margin for three dimensional conformal radiotherapy (3D CRT) of prostate cancer, the uncertainties in the internal positions of the prostate and seminal vesicles (SV) and in the treatment setups were measured. METHODS AND MATERIALS: Weekly computed tomography (CT) scans of the pelvis (n=51) and daily electronic portal images (n=1630) were reviewed for eight patients who received seven-field 3D CRT for prostate cancer. The CT scans were registered in three dimensions to the original planning CT scan using commercially available software to measure the center-of volume (COV) motion of the prostate and SV. The daily portal images were registered to the corresponding simulation films to measure the setup displacements. The standard deviation (SD) of the internal organ motions was added to the SD of the setups in quadrature to determine the total uncertainty. Positive directions were left, anterior, and superior. Rotations necessary to register the CT scans and portal images were minimal and not further analyzed. RESULTS: The mean motion for the COV of the prostate+/-the SD was 0+/-0.9 mm in the left-right (LR), 0.5+/-2.6 mm in the anterior-posterior (AP), and 1.5+/-3.9 mm in the superior-inferior (SI) directions. The mean motion for the COV of the SV+/-the SD was 0.3+/-1.7 mm in the LR, 0.7+/-3.8 mm in the AP, and 0.9+/-3.5 mm in the SI directions. For all patients the mean isocenter displacement+/-the SD was 0+/-3.1 mm in the LR, 1.4+/ 3.0 mm in the AP, and -0.4+/-2.1 mm in the SI directions. The total uncertainty for the prostate was 3.2 mm, 4.0 mm, and 4.4 mm in the LR, AP, and SI directions, respectively. For the SV, the total uncertainty was 3.5, 4.8, and 4.1 mm in the LR, AP, and SI directions, respectively. CONCLUSIONS: PTV margins of 10 to 16 mm are required to encompass all (99%) possible positions of the prostate or SV during 3D CRT. PTV margins of 7 to 11 mm will encompass the measured uncertainties with a 95% probability. PTV margins of 5 mm may not adequately cover the intended volume. PMID- 9747841 TI - Radiation therapy port films: a quality assurance study. AB - PURPOSE: The purpose of this study was to assess the port film acceptance rate in a large community practice setting and to catalog the reasons for rejection. METHODS: Between December 1993 and July 1996, a quality assurance monitor log was maintained on 4,150 patients who underwent a total of 4,450 treatment courses. Port films were taken at the beginning and at the half way point in the treatment course. A total of 20,735 port films were compared with the matching simulation films. We recorded the site being treated, the radiation oncologist who reviewed the films and the reason for rejection. RESULTS: The monthly acceptance rate varied from a low of 67% to a high of 83%, with a gradual upward trend. The single most common reason for rejecting films was a centering problem-12% of all films taken were rejected for this reason. The next most common problems were block placement or body setup errors that caused 3.4% and 2.7% of the films to be rejected, respectively. Average acceptance rates between 10 different sites (abdomen, brain, breast, chest, extremities, head and neck, pelvis, prostate, rectum and spine) varied from 68% to 80%. Individual differences between 12 radiation oncologists reviewing the films varied from 67% to 87%. CONCLUSIONS: A detailed analysis of field localization errors allowed us to identify areas where improvement was needed and suggested that specific guidelines for acceptance would help reduce the variability noted in the acceptance rate between sites and physicians. PMID- 9747842 TI - Whole-body dose from tomotherapy delivery. AB - PURPOSE: To measure whole-body dose in tomotherapy of the head and neck region resulting from internal patient scatter and linear accelerator leakage. METHODS AND MATERIALS: Treatments are performed using a commercial computer-controlled intensity modulated radiation therapy planning and delivery system (Peacock, NOMOS Corp.) and a 6-MV linear accelerator (Clinac 6/100, Varian Corp.). The patient dose outside the treatment field is measured in a water-equivalent phantom using thermoluminescent dosimetry. The whole-body dose components from internal scatter and leakage are separately determined. The use of fixed-portal leakage and scattered radiation measurements to estimate the whole-body dose from tomotherapy is evaluated. RESULTS: The internally scattered dose is significant near the target, but becomes negligible relative to the leakage dose beyond 15 cm from the target. Dose at 10 cm from the target volume, due to internal scatter and leakage, is approximately 2.5% of the total target dose, reducing to 0.5% at 30 cm. The measured dose is relatively uniform throughout the phantom. CONCLUSION: The whole-body dose equivalent from a tomotherapy treatment is greater than that from conventional radiation therapy. Further studies are required to assess the trade-off between improved dose distribution conformality and a possible slight increase in radiation-induced fatal malignancies. The accuracy of using fixed-portal leakage and scattered dose measurements to estimate the whole-body dose from tomotherapy treatments is adequate, if the appropriate fixed-portal field size equivalent is used. PMID- 9747843 TI - The sugarcane bacilliform badnavirus promoter is active in both monocots and dicots. AB - Regions of the sugarcane bacilliform badnavirus genome were tested for promoter activity. The genomic region spanning nucleotides 5999-7420 was shown to possess promoter activity as exemplified by its ability to drive the expression of the coding region of the uidA gene of Escherichia coli, in both Avena sativa and Arabidopsis thaliana. In A. sativa, the promoter was active in all organs examined and, with the exception of the anthers where the expression was localized, this activity was constitutive. In A. thaliana, the promoter activity was constitutive in the rosette leaf, stem, stamen, and root and limited primarily to vascular tissue in the sepal and the silique. The transgene was inherited and active in progeny plants of both A. sativa and A. thaliana. PMID- 9747844 TI - Ribonucleoprotein particles of quiescent maize embryonic axes. AB - Certain RNA molecules are known to be sequestered and stored as ribonucleoprotein particles (RNPs) in many different tissues, particularly at some stages of metabolic quiescence. In this research RNPs from embryonic axes of mature maize seeds were isolated by sucrose and CsCl gradient centrifugation and characterized based on their RNA and protein contents. Two types of RNP particles of non ribosomal nature were identified by northern blot analysis with specific probes: the 7S RNP and the signal recognition particle (SRP) particles which contain 5S rRNA and 7S RNA respectively. The proteins associated to these RNA molecules were the transcription factor TFIIIA-homologous protein associated to 7S RNP, and the p72, p68 and p54-GTPase proteins associated to SRP. PMID- 9747845 TI - Amino-terminal and hydrophobic regions of the Chlamydomonas reinhardtii plastocyanin transit peptide are required for efficient protein accumulation in vivo. AB - Nucleus-encoded chloroplast proteins of vascular plants are synthesized as precursors and targeted to the chloroplast by stroma-targeting domains in N terminal transit peptides. Transit peptides in Chlamydomonas reinhardtii are considerably shorter than those in vascular plants, and their stroma-targeting domains have similarities to both mitochondrial and chloroplast targeting sequences. To examine Chlamydomonas transit peptide function in vivo, deletions were introduced into the transit peptide coding region of the petE gene, which encodes the thylakoid lumen protein plastocyanin (PC). The mutant petE genes were introduced into a plastocyanin-deficient Chlamydomonas strain, and transformants that accumulated petE mRNA were analyzed for PC accumulation. The most profound defects were observed with deletions at the N-terminus and those that extended into the hydrophobic region in the C-terminal half of the transit peptide. PC precursors were detected among pulse-labeled proteins in transformants with N terminal deletions, suggesting that these precursors cannot be imported and are degraded in the cytosol. Intermediate PC species were observed in a transformant deleted for part of the hydrophobic region, suggesting that this protein is defective in lumen translocation and/or processing. Thus, despite its shorter length, the bipartite nature of the Chlamydomonas PC transit peptide appears similar to that of lumen-targeted proteins in vascular plants. Analysis of the synthesis, stability, and accumulation of PC species in transformants bearing deletions in the stroma-targeting domain suggests that specific regions probably have distinct roles in vivo. PMID- 9747846 TI - Expression of enzymatically active, recombinant barley alpha-glucosidase in yeast and immunological detection of alpha-glucosidase from seed tissue. AB - An alpha-glucosidase cDNA clone derived from barley aleurone tissue was expressed in Pichia pastoris and Escherichia coli. The gene was fused with the N-terminal region of the Saccharomyces cerevisiae alpha-factor secretory peptide and placed under control of the Pichia AOX1 promoter in the vector pPIC9. Enzymatically active, recombinant alpha-glucosidase was synthesized and secreted from the yeast upon induction with methanol. The enzyme hydrolyzed maltose > trehalose > nigerose > isomaltose. Maltase activity occurred over the pH range 3.5-6.3 with an optimum at pH 4.3, classifying the enzyme as an acid alpha-glucosidase. The enzyme had a Km of 1.88 mM and Vmax of 0.054 micromol/min on maltose. The recombinant alpha-glucosidase expressed in E. coli was used to generate polyclonal antibodies. The antibodies detected 101 and 95 kDa forms of barley alpha-glucosidase early in seed germination. Their levels declined sharply later in germination, as an 81 kDa alpha-glucosidase became prominent. Synthesis of these proteins also occurred in isolated aleurones after treatment with gibberellin, and this was accompanied by a 14-fold increase in alpha-glucosidase enzyme activity. PMID- 9747847 TI - Cre/lox-mediated site-specific integration of Agrobacterium T-DNA in Arabidopsis thaliana by transient expression of cre. AB - The Cre/lox system was used to obtain targeted integration of an Agrobacterium T DNA at a lox site in the genome of Arabidopsis thaliana. Site-specific recombinants, and not random events, were preferentially selected by activation of a silent lox-neomycin phosphotransferase (nptII) target gene. To analyse the effectiveness of Agrobacterium-mediated transfer we used T-DNA vectors harbouring a single lox sequence (this vector had to circularize at the T-DNA left- and right-border sequences prior to site-specific integration) or two lox sequences (this vector allowed circularization at the lox sequences within the T-DNA either prior to or after random integration, followed by targeting of the circularized vector), respectively. Furthermore, to control the reversibility of the integration reaction, Cre recombinase was provided transiently by using a cotransformation approach. One precise stable integrant was found amongst the recombinant calli obtained after transformation with a double-lox T-DNA vector. The results indicate that Agrobacterium-mediated transformation can be used as a tool to obtain site-specific integration. PMID- 9747848 TI - Use of alternate splice sites in granule-bound starch synthase mRNA from low amylose rice varieties. AB - The rice Waxy gene encodes a granule-bound starch synthase (GBSS) necessary for the synthesis of amylose in endosperm tissue. We have previously shown that a CT microsatellite near the transcriptional start site of the GBSS gene can distinguish 7 alleles that accounted for more than 80% of the variation in apparent amylose content in an extended pedigree of 89 US rice cultivars (Oryza sativa L.). Furthermore, all the cultivars with 18% or less amylose were shown to have the sequence AGTTATA at the putative leader intron 5' splice site, while all cultivars with a higher proportion of amylose had AGGTATA. Here we demonstrate that this single-base mutation reduces the efficiency of GBSS pre-mRNA processing and results in alternate splicing at three cryptic sites. The predominant 5' splice site in CT18 low-amylose varieties is 93 bp upstream of the splice site used in intermediate and high amylose varieties and is immediately 5' to the CT microsatellite that we previously demonstrated to be tightly correlated with amylose content. Use of the leader intron 5' splice site at either -93 or -1 in conjunction with the predominant 3' splice site results in formation of a small open reading frame 38 bp upstream of the normal ATG and out of frame with it. This open reading frame is not produced when any of the 5' leader intron splice sites are used in conjunction with an alternate 3' splice site five bases further downstream which was observed in all rice varieties tested. PMID- 9747849 TI - A ca. 40 kDa maize (Zea mays L.) embryo dehydrin is encoded by the dhn2 locus on chromosome 9. AB - Dehydrins (LEA D11 proteins) are the products of multigene families in a number of higher plants. To date, however, only one dehydrin locus, dhn1 (a major embryo and drought-induced protein of ca. 18 kDa) has been placed on chromosome 6L of the genetic linkage map of maize. The presence of a larger, ca. 40 kDa embryo protein that is also specifically detected by anti-dehydrin antibodies had been observed in some maize inbreds, including B73, suggesting that other dhn loci may exist. The ca. 22 kDa and ca. 40 kDa immunopositive proteins were purified from B73 and their amino acid compositions determined. The two proteins' amino acid compositions are typical of dehydrins, yet they differ from each other, indicating that they are distinct dhn gene products. Different size alleles for both proteins, or presence/absence in the case of the ca. 40 kDa protein, were evident from comparisons of embryo proteins of various maize inbreds. Analysis of segregating F2 progeny derived from self-pollination of F1 hybrids from four crosses (B73 x OH43, Mo17 x A632, AHO x A632, Latente x A632) revealed that alleles of the two genes assort independently. Map positions of the two dhn loci were then determined using two maize recombinant inbred line (RIL) mapping populations. The predicted map position of the gene controlling production of the ca. 22 kDa protein confirmed that this protein is the product of the dhn1 gene. The gene encoding the ca. 40 kDa dehydrin-like protein maps to a new locus on chromosome 9S near wx1, which we have named dhn2. PMID- 9747850 TI - Expression analysis of a sucrose carrier in the germinating seedling of Ricinus communis. AB - This study describes the expression of a sucrose carrier at various developmental stages in Ricinus communis. A partial-length cDNA clone, RcSUT1, was isolated by RT-PCR from Ricinus seedling RNA. This is almost identical to a sucrose carrier cDNA, Rscr1, which has previously been isolated by library screening. However, we have observed a very different expression pattern in the seedling to that previously reported. Northern analysis, with RcSUT1 as a probe, revealed high expression of a 2 kb transcript in the cotyledons of the germinating seedling; transcript levels were similar in cotyledons harvested 3-6 days after germination. A much lower level of this transcript was detected in the root, hypocotyl and endosperm RNA of the seedling and very low levels were also present in the sink and source leaves of the mature plant. This pattern of expression was also reflected at the protein level with an antipeptide antibody raised to part of the RcSUT1 deduced amino acid sequence. Tissue print hybridisation analysis of the hypocotyl revealed that the sucrose carrier transcripts were localised to the phloem cells of the vascular bundles. A more detailed analysis of sucrose carrier gene expression in the cotyledons of the germinating seedling was carried out by in situ hybridisation; the strongest signals were observed from the lower epidermal layer and the phloem, consistent with an active loading role for these cells. An ultrastructural study of the cells in the lower epidermis showed that they have wall ingrowths which are characteristic of transfer cells. The results are discussed in relation to the physiological role of the sucrose carrier in the Ricinus seedling and to the pathways of sucrose movement from endosperm to the sieve elements in the cotyledons. PMID- 9747851 TI - Molecular cloning of an Arabidopsis cDNA encoding a dynamin-like protein that is localized to plastids. AB - Dynamin-related proteins are high molecular weight GTPase proteins found in a variety of eukaryotic cells from yeast to human. They are involved in diverse biological processes that include endocytosis in animal cells and vacuolar protein sorting in yeast. We isolated a new gene, ADL2, that encodes a dynamin like protein in Arabidopsis. The ADL2 cDNA is 2.68 kb in size and has an open reading frame for 809 amino acid residues with a calculated molecular mass of 90 kDa. Sequence analysis of ADL2 revealed a high degree of amino acid sequence similarity to other members of the dynamin superfamily. Among those members ADL2 was most closely related to Dnm1p of yeast and thus appears to be a member of the Vps1p subfamily. Expression studies showed that the ADL2 gene is widely expressed in various tissues with highest expression in flower tissues. In vivo targeting experiments showed that ADL2:smGFP fusion protein is localized to chloroplasts in soybean photoautroph cells. In addition experiments with deletion constructs revealed that the N-terminal 35 amino acid residues were sufficient to direct the smGFP into chloroplasts in tobacco protoplasts when expressed as a fusion protein. PMID- 9747852 TI - Apple ACC-oxidase and polygalacturonase: ripening-specific gene expression and promoter analysis in transgenic tomato. AB - Levels of 1-aminocyclopropane-1-carboxylate (ACC) oxidase and polygalacturonase (PG) mRNAs were characterized during ripening of Royal Gala, Braeburn and Granny Smith apples. Both ACC-oxidase and PG mRNAs were up-regulated in ripening fruit of all three cultivars. Expression in Royal Gala was detected earlier than in Braeburn and Granny Smith, relative to internal ethylene concentration. Genomic clones corresponding to the ACC-oxidase and PG mRNAs expressed in ripe apple fruit were isolated and ca. 2 kb of each promoter was sequenced. The start point of transcription in each gene was mapped by primer extension, and sequences homologous to elements in other ethylene-responsive or PG promoters were identified. The fruit specificity of the apple ACC-oxidase and PG promoters was investigated in transgenic tomato plants using a nested set of promoter fragments fused to the beta-glucuronidase (gusA) reporter gene. For the ACC-oxidase gene, 450 bp of 5' promoter sequence was sufficient to drive GUS expression, although this expression was not specific to ripening fruit. Larger fragments of 1966 and 1159 bp showed both fruit and ripening specificity. For the PG gene, promoter fragments of 1460 and 532 bp conferred ripening-specific expression in transgenic tomato fruit. However GUS expression was down-regulated by 2356 bp of promoter, suggesting the presence of a negative regulatory element between positions -1460 and -2356. PMID- 9747853 TI - Retrotransposon-like sequences integrated into the genome of pineapple, Ananas comosus. AB - Retrotransposon-like sequences have been serendipitously detected in the genome of commercial pineapple, Ananas comosus. The sequence from a 2.6 kb cloned fragment of this element had greatest similarity to the del1 Lilium henryi retrotransposon and the gypsy/Ty3 group of retroelements. The order of the genes from 5' to 3' was reverse transcriptase, ribonuclease H and integrase. The integrase domain contained the amino acid sequence motifs which have been associated with recognition of the long terminal repeats and with the cutting/joining reactions required for integration of similar retroelements into the host genome. The retrotransposon existed as a population of variable sequences which were dispersed throughout the genome of pineapple. Southern hybridisation showed that the retrotransposon had integrated repeatedly into the pineapple genome. The reading frame of the element was not interrupted by stop codons, suggesting that it is still potentially capable of transposing. This is the first report of a retrotransposon in pineapple, which we have called deal (for dispersed element of Ananas). PMID- 9747854 TI - Expression of antisense chalcone synthase RNA in transgenic hybrid walnut microcuttings. Effect on flavonoid content and rooting ability. AB - Walnut somatic embryos (Juglans nigra x Juglans regia) were transformed with a vector containing a neomycin phosphotransferase II, a beta-glucuronidase and an antisense chalcone synthase (chs) gene. This antisense construct included a 400 bp cDNA fragment of a walnut chs gene under the control of the duplicated CaMV 35S promoter. Molecular, biochemical and biological characterizations were performed both on transformed embryos propagated by secondary somatic embryogenesis and on microshoots developed by in vitro culture of embryonic epicotyls from somatic embryos. Thirteen transformed lines with the vector containing the antisense chs gene, one line with only the gus and nptII genes and one untransformed line were maintained in tissue culture. Six of the antisense lines were shown to be flavonoid-deficient. They exhibited a strongly reduced expression of chs genes, very low chalcone synthase activity and no detectable amounts of quercitrin, myricitrin, flavane-3-ols and proanthocyanidins in stems. Rooting tests showed that decreased flavonoid content in stems of antisense chs transformed lines was associated with enhanced adventitious root formation. Free auxin and conjugated auxin contents were determined during the latter phase of the micropropagation, and no variations were detected between control and antisense chs transformed lines. The in vitro plants developed a large basal callus and apical necrosis upon auxinic induction and the transformed lines highly deficient in flavonoids were more sensitive to exogenous application of indolebutyric acid (IBA). PMID- 9747855 TI - Tissue- and stage-specific expression of a soybean (Glycine max L.) seed maturation, biotinylated protein. AB - A cDNA clone GmPM4 which encodes mRNA species in mature or dry soybean seeds was characterized. DNA sequence analysis shows that the deduced polypeptides have a molecular mass of 68 kDa. GmPM4 proteins have a relatively high amino acid sequence homology with a major biotinylated protein isolated from pea seeds, SBP65, but both of these proteins differ markedly from that of presently known biotin enzymes. The accumulation of GmPM4 mRNA is detectable in the leaf primodium and the vascular tissues of the hypocotyl-radicle axis of mature seeds, and the GmPM4 proteins are present at high levels in dry and mature soybean seeds, but not in fresh immature seeds. It degrades rapidly at the early stage of seed germination. These proteins are boiling-soluble and biotinylated when they are present endogenously in soybean seeds; however, the same recombinant protein expressed in Escherichia coli is boiling-soluble, but it is not biotinylated. PMID- 9747856 TI - Processing of a chimeric protein in chloroplasts is different in transgenic maize and tobacco plants. AB - Transgenic maize (Zea mays L.) and tobacco (Nicotiana tabacum Petit Havana SR1) plants have been generated, which overproduce a mitochondrial Nicotiana plumbaginifolia manganese superoxide dismutase (MnSOD) in chloroplasts. For this, the mature MnSOD-coding sequence was fused to a chloroplast transit peptide from a Pisum sativum ribulose-1,5-bisphosphate carboxylase (Rubisco) gene and expression of the chimeric gene was driven by the cauliflower mosaic virus (CaMV) 35S promoter. The transgenic MnSOD gene product was correctly targeted to the chloroplasts both in maize and tobacco. However, despite the use of the CaMV 35S promoter, the MnSOD was predominantly localized in the chloroplasts of the bundle sheath cells of maize. Furthermore, the transit peptide was cleaved off at a different position in maize and tobacco. PMID- 9747857 TI - Maize mitochondrial seryl-tRNA synthetase recognizes Escherichia coli tRNA(Ser) in vivo and in vitro. AB - In our studies to analyze the structure/function relationships among cytoplasmic and organellar seryl-tRNA synthetases (SerRS), we have characterized a Zea mays cDNA (SerZMm) encoding a protein with significant similarity to prokaryotic SerRS enzymes. To demonstrate the functional identity of SerZMm, the gene sequence encoding the putative mature protein was cloned. This construct complemented in vivo a temperature-sensitive Escherichia coli serS mutant strain. The mature SerZMm protein overexpressed in Escherichia coli efficiently aminoacylated bacterial tRNA(Ser) in vitro, while yeast tRNA was a poor substrate. These data identify SerZMm as an organellar maize seryl-tRNA synthetase, the first plant organellar SerRS to be cloned. The analysis of its N-terminal targeting signal suggests a mitochondrial function for the SerZMm protein in maize. PMID- 9747858 TI - Should selection of adjuvant chemotherapy for patients with breast cancer be based on erbB-2 status? PMID- 9747860 TI - Biotechnology industry roars into the millennium. PMID- 9747859 TI - Is there a downside to elderly women undergoing screening mammography? PMID- 9747861 TI - New Pap test technologies hit heavy seas but sales keep flying. PMID- 9747862 TI - Bisphosphonates: lingering questions about their use. PMID- 9747863 TI - FDA assigns drug master file to European cancer group. PMID- 9747864 TI - New computer program assesses a woman's risk for developing breast cancer. PMID- 9747865 TI - Second lung cancers in patients after treatment for an initial lung cancer. AB - BACKGROUND: Prospectively and retrospectively identified patient cohorts that were successfully treated for primary lung cancer have been followed to document the rate of development of and the effectiveness of treatment of second lung cancers. This review was performed to assess rates of second lung cancer development, factors associated with the development of these cancers, and the success of their treatment. METHODS: The MEDLINE database was searched to identify articles published in English concerning lung cancers, second primary cancers, treatment of these cancers, and patient survival. RESULTS: The risk of developing a second lung cancer in patients who survived resection of a non-small cell lung cancer is approximately 1%-2% per patient per year. Approximately one half of the patients who develop second non-small-cell lung cancers can have these tumors resected. The median survival from diagnosis of a second lung cancer in these patients is between 1 and 2 years, with a 5-year survival of approximately 20% (range, 4%-32%). The average risk of developing a second lung cancer in patients who survived small-cell lung cancer is approximately 6% per patient per year. For patients who survived small-cell cancer, the risk increases from approximately 2% to greater than 10% per patient per year 10 years after initial treatment. Only 7% (range, 6%-12%) of patients treated for small-cell lung cancer survive 2 years or more. Survivors who continue to smoke cigarettes have an increased risk of developing a second lung cancer. CONCLUSIONS: In patients surviving an initial lung cancer, the cumulative risk for the development of a second primary lung cancer makes this cancer a common cause of death. The high risk of developing a second lung cancer makes patients with these cancers an important population for study of surveillance strategies and chemoprevention agents. PMID- 9747866 TI - erbB-2, p53, and efficacy of adjuvant therapy in lymph node-positive breast cancer. AB - BACKGROUND: We have previously reported that high expression of the erbB-2 gene (also known as HER-2/neu and ERBB2) in breast cancer is associated with patient response to dose-intensive treatment with cyclophosphamide, doxorubicin (Adriamycin), and 5-flurouracil (CAF) on the basis of short-term follow-up of 397 patients (set A) with axillary lymph node-positive tumors who were enrolled in Cancer and Leukemia Group B (CALGB) protocol 8541. METHODS: To validate those findings, we conducted immunohistochemical analyses of erbB-2 and p53 protein expression in an additional cohort of 595 patients (set B) from CALGB 8541, as well as a molecular analysis of erbB-2 gene amplification in tumors from all patients (sets A and B). Marker data were compared with clinical, histologic, treatment, and outcome data. RESULTS: Updated analyses of data from set A (median follow-up, 10.4 years) showed an even stronger interaction between erbB-2 expression and CAF dose, by use of either immunohistochemical or molecular data. A similar interaction between erbB-2 expression and CAF dose was observed in all 992 patients, analyzed as a single group. However, for set B alone (median follow up, 8.2 years), results varied with the method of statistical analysis. By use of a proportional hazards model, the erbB-2 expression-CAF dose interaction was not significant for all patients. However, in the subgroups of patients randomly assigned to the high- or the moderate-dose arms, significance was achieved. When patient data were adjusted for differences by use of a prognostic index (to balance an apparent failure of randomization in the low-dose arm), the erbB-2 expression-CAF dose interaction was significant in all patients from the validation set B as well. An interaction was also observed between p53 immunopositivity and CAF dose. CONCLUSIONS: The hypothesis that patients whose breast tumors exhibit high erbB-2 expression benefit from dose-intensive CAF should be further validated before clinical implementation. Interactions between erbB-2 expression, p53 expression, and CAF dose underscore the complexities of predictive markers where multiple interactions may confound the outcome. PMID- 9747867 TI - erbB-2 and response to doxorubicin in patients with axillary lymph node-positive, hormone receptor-negative breast cancer. AB - BACKGROUND: Overexpression of the erbB-2 protein by breast cancer cells has been suggested to be a predictor of response to doxorubicin. A retrospective study was designed to test this hypothesis. METHODS: In National Surgical Adjuvant Breast and Bowel Project protocol B-11, patients with axillary lymph node-positive, hormone receptor-negative breast cancer were randomly assigned to receive either L-phenylalanine mustard plus 5-fluorouracil (PF) or a combination of L phenylalanine mustard, 5-fluorouracil, and doxorubicin (PAF). Tumor cell expression of erbB-2 was determined by immunohistochemistry for 638 of 682 eligible patients. Statistical analyses were performed to test for interaction between treatment and erbB-2 status (positive versus negative) with respect to disease-free survival (DFS), survival, recurrence-free survival (RFS), and distant disease-free survival (DDFS). Reported P values are two-sided. RESULTS: Overexpression of erbB-2 (i.e., positive immunohistochemical staining) was observed in 239 (37.5%) of the 638 tumors studied. Overexpression was associated with tumor size (P=.02), lack of estrogen receptors (P=.008), and the number of positive lymph nodes (P=.0001). After a mean time on study of 13.5 years, the clinical benefit from doxorubicin (PAF versus PF) was statistically significant for patients with erbB-2-positive tumors--DFS: relative risk of failure (RR)=0.60 (95% confidence interval [CI]=0.44-0.83), P=.001; survival: RR=0.66 (95% CI=0.47 0.92), P =.01; RFS: RR=0.58 (95% CI=0.42-0.82), P=.002; DDFS: RR=0.61 (95% CI=0.44-0.85), P=.003. However, it was not significant for patients with erbB-2 negative tumors-DFS: RR=0.96 (95% CI=0.75-1.23), P=.74; survival: RR =0.90 (95% CI=0.69-1.19), P=.47; RFS: RR=0.88 (95% CI=0.67-1.16), P=.37; DDFS: RR=1.03 (95% CI=0.79-1.35), P=.84. Interaction between doxorubicin treatment and erbB-2 overexpression was statistically significant for DFS (P=.02) and DDFS (P=.02) but not for survival (P= .15) or RFS (P=.06). CONCLUSIONS: These data support the hypothesis of a preferential benefit from doxorubicin in patients with erbB-2 positive breast cancer. PMID- 9747868 TI - Tamoxifen for prevention of breast cancer: report of the National Surgical Adjuvant Breast and Bowel Project P-1 Study. AB - BACKGROUND: The finding of a decrease in contralateral breast cancer incidence following tamoxifen administration for adjuvant therapy led to the concept that the drug might play a role in breast cancer prevention. To test this hypothesis, the National Surgical Adjuvant Breast and Bowel Project initiated the Breast Cancer Prevention Trial (P-1) in 1992. METHODS: Women (N=13388) at increased risk for breast cancer because they 1) were 60 years of age or older, 2) were 35-59 years of age with a 5-year predicted risk for breast cancer of at least 1.66%, or 3) had a history of lobular carcinoma in situ were randomly assigned to receive placebo (n=6707) or 20 mg/day tamoxifen (n=6681) for 5 years. Gail's algorithm, based on a multivariate logistic regression model using combinations of risk factors, was used to estimate the probability (risk) of occurrence of breast cancer over time. RESULTS: Tamoxifen reduced the risk of invasive breast cancer by 49% (two-sided P<.00001), with cumulative incidence through 69 months of follow-up of 43.4 versus 22.0 per 1000 women in the placebo and tamoxifen groups, respectively. The decreased risk occurred in women aged 49 years or younger (44%), 50-59 years (51%), and 60 years or older (55%); risk was also reduced in women with a history of lobular carcinoma in situ (56%) or atypical hyperplasia (86%) and in those with any category of predicted 5-year risk. Tamoxifen reduced the risk of noninvasive breast cancer by 50% (two-sided P<.002). Tamoxifen reduced the occurrence of estrogen receptor-positive tumors by 69%, but no difference in the occurrence of estrogen receptor-negative tumors was seen. Tamoxifen administration did not alter the average annual rate of ischemic heart disease; however, a reduction in hip, radius (Colles'), and spine fractures was observed. The rate of endometrial cancer was increased in the tamoxifen group (risk ratio = 2.53; 95% confidence interval = 1.35-4.97); this increased risk occurred predominantly in women aged 50 years or older. All endometrial cancers in the tamoxifen group were stage I (localized disease); no endometrial cancer deaths have occurred in this group. No liver cancers or increase in colon, rectal, ovarian, or other tumors was observed in the tamoxifen group. The rates of stroke, pulmonary embolism, and deep-vein thrombosis were elevated in the tamoxifen group; these events occurred more frequently in women aged 50 years or older. CONCLUSIONS: Tamoxifen decreases the incidence of invasive and noninvasive breast cancer. Despite side effects resulting from administration of tamoxifen, its use as a breast cancer preventive agent is appropriate in many women at increased risk for the disease. PMID- 9747869 TI - Diagnostic testing following screening mammography in the elderly. AB - BACKGROUND: To provide some sense of the general frequency and timing of diagnostic testing following screening mammography in the United States, we investigated the experience of women screened in the Medicare population. METHODS: By use of Medicare's National Claims History System, we identified a cohort (n=23172) of women 65 years old or older screened during the period from January 1, 1995, through April 30, 1995, and tracked each woman over the subsequent 8 months for the performance of additional breast imaging and biopsy procedures. Using two claims-based definitions for newly detected breast cancer, we also estimated the positive predictive value of screening mammography. RESULTS: For every 1000 women aged 65-69 years who underwent screening, 85 (95% confidence interval [CI]=79-91) had follow-up testing in the subsequent 8 months; 76 (95% CI=71-82) had additional breast imaging, and 23 (95% CI=20-26) had biopsy procedures. Corresponding numbers for women aged 70 years or more were similar. Some women underwent repeated examinations; 13% of those receiving diagnostic mammograms had more than one; 11% of those undergoing biopsy procedures had more than one. About half of the women who underwent a biopsy had the procedure more than 3 weeks after the imaging test upon which the decision to perform a biopsy was presumably made. The estimated positive predictive value of an abnormal screening mammogram (defined as a mammogram that engendered additional testing) was 0.08 (95% CI=0.06-0.10) for women aged 65-69 years and 0.14 (95% CI=0.12 0.16) for women aged 70 years or more. CONCLUSION: Additional testing is a frequent consequence of screening mammography and may require a considerable period of time to come to closure. The need for additional testing, however, is weakly predictive of cancer. PMID- 9747870 TI - Benzo[a]pyrene diol epoxide and bleomycin sensitivity and susceptibility to cancer of upper aerodigestive tract. AB - BACKGROUND: Tobacco smoking is an established risk factor for cancers of the upper aerodigestive tract, and measurement of chromosomal aberrations, i.e., chromatid breaks, induced in lymphocytes in vitro by bleomycin has been shown to be a predictor of risk for these cancers. In a case-control study, we recruited case subjects who were previously treated with surgery and/or radiotherapy for stage I or stage II squamous cell carcinoma of the head and neck to test the hypothesis that lymphocytic chromatid breaks induced by benzo[a]pyrene diol epoxide (BPDE), a tobacco mutagen, may also be associated with risk of developing cancers of the upper aerodigestive tract. METHODS: Case subjects were matched to control subjects on the basis of age, sex, ethnicity, and smoking status. Primary lymphocytes from 67 case subjects and 81 control subjects were treated with 2 microM BPDE for 24 hours, and the frequency of induced chromatid breaks was determined. All statistical tests were two-sided. RESULTS: Lymphocytes from case subjects compared with lymphocytes from control subjects showed significantly more breaks per cell induced by BPDE (mean+/-standard deviation, 0.77+/-0.38 versus 0.49+/-0.25; P<.001). Lymphocytes from 64.2% of case subjects were sensitive to BPDE (using a cutoff value of > or =0.60 break per cell). Subjects in the highest quartile of chromatid breaks had an approximately 20-fold increased risk of cancer compared with those in the lowest quartile after adjustment for age, sex, ethnicity, and smoking status. The association between BPDE sensitivity and cancer risk was higher in former smokers than in current smokers and higher in younger patients than in older patients. Subjects with sensitivity to both BPDE and bleomycin were at a 19.2-fold increased risk of cancer compared with those who were not sensitive to either agent. CONCLUSIONS: Mutagen sensitivity assays may aid in identifying individuals at risk of cancer, and use of parallel assays with two mutagens may improve risk predictability. PMID- 9747871 TI - Potentiation of lonidamine and diazepam, two agents acting on mitochondria, in human glioblastoma treatment. AB - BACKGROUND: Cellular metabolism in glioblastoma multiforme, the most common primary brain tumor in humans, is characterized by a high rate of aerobic glycolysis that is dependent on mitochondria-bound hexokinase. Moreover, high levels of glucose utilization and tumor aggressiveness in glioblastoma are associated with a high density of mitochondrial benzodiazepine receptors. We sought to inhibit glioblastoma metabolism by simultaneously inhibiting hexokinase with lonidamine and binding benzodiazepine receptors with diazepam. METHODS: Cellular glioblastoma metabolism in five glioblastoma cell lines was assessed in vitro by measuring cell proliferation (by use of a tetrazolium-based colorimetric assay, measurement of DNA synthesis, and assessment of cell cycle distribution), by measuring membrane fluidity (by fluorescence polarization measurement of cells stained with a fluorescent probe), and by measuring changes in intracellular pH. Immunodeficient nude mice bearing subcutaneous xenografts of human glioblastoma cells were used to assess the antitumor activities of lonidamine and diazepam; the mice were treated twice daily with lonidamine (total daily dose of 160 mg/kg body weight) and/or diazepam (total daily dose of 1 mg/kg body weight) for 10 consecutive days. RESULTS: When used in combination, the two drugs had a stronger effect on glioblastoma cell proliferation and metabolism in vitro than did either agent used alone. In vivo, the combination of lonidamine and diazepam was significantly more effective in reducing glioblastoma tumor growth than either drug alone (two-sided P<.01, Mann-Whitney U test, comparing growth of treated tumors with that of untreated tumors); this tumor growth retardation was maintained as long as treatment was given. CONCLUSION: The combination of lonidamine and diazepam--drugs that target two distinct mitochondrial sites involved in cellular energy metabolism--potentiates the effects of the individual drugs and may prove useful in the treatment of human glioblastomas. PMID- 9747872 TI - Resistance to Fas-mediated apoptosis: activation of caspase 3 is regulated by cell cycle regulator p21WAF1 and IAP gene family ILP. AB - The death receptor Fas transduces apoptotic death signaling mediated by caspases. In the present study, human hepatoma HepG2 cells showed the Fas-mediated apoptosis mediated by caspase, especially caspase 3, only in the presence of actinomycin D. Interestingly, cytosolic proteins extracted from intact HepG2 cells induced caspase 3 inactivation. Our results reveal that this inactivation was triggered by the direct inhibition of activated caspase 3 by IAP gene family ILP. In addition, a 53 kDa protein was co-immunoprecipitated with anti-human caspase 3 antibody from intact HepG2 cells. This protein was a complex-protein of procaspase 3 and the cell cycle regulator p21WAF1 (p21). P21 bound to only procaspase 3, but not to activated caspase 3. We also demonstrate that p21 protein-loaded HepG2 cells resist to Fas-mediated apoptosis even in the presence of actinomycin D. Here we report that caspase 3 inactivation for the resistance to Fas-mediated apoptosis is induced by a procaspase 3/p21 complex formation and direct inhibition of activated caspase 3 by ILP. PMID- 9747873 TI - The SH2-containing adapter protein GRB10 interacts with BCR-ABL. AB - Bcr-Abl is an oncogenic tyrosine kinase expressed in tumor cells of CML and a subset of ALL which in its unregulated and activated state is thought to cause cell transformation and leukemia. Bcr-Abl contains several autophosphorylation sites which serve as potential docking sites for SH2-containing signaling molecules. Mutational analysis has indicated that these autophosphorylation sites play a critical role in the transforming capability of Bcr-Abl. It has been shown that the SH2-containing adapter protein Grb2 binds to the autophosphorylation site Tyr(p)177 whereby it couples Bcr-Abl to the Ras pathway. The biological consequences of this interaction, however, are presently unclear. A Tyr177 mutated Bcr-Abl which lacks the ability to interact with the Grb2-SH2 domain still transforms myeloid cells and generates tumors in nude mice. We performed a yeast two-hybrid screen to identify signaling proteins which bind to distinct Bcr Abl autophosphorylation sites. Autophosphorylation of Bcr-Abl in yeast was accomplished by using the DNA binding protein LexA which permits dimerization and crossphosphorylation of the fused bait. Using a LexA-Bcr-Abl full length fusion protein as bait, we identified several SH2-containing proteins. Among them we confirmed molecules already shown by others to interact with Bcr-Abl, in vivo, including Grb2, PI-3-kinase and Crk indicating that dimerization in yeast leads to autophosphorylation of tyrosine residues crucial for Bcr-Abl signaling in vivo. More importantly, we identified the SH2-containing protein Grb10 as a new binding partner for Bcr-Abl. This binding occurs in a phosphotyrosine-dependent manner at Bcr sites of Bcr-Abl. Both Abl and Bcr alone, as well as a kinase defective Bcr-Abl, failed to interact with Grb10 in yeast. Mutational analysis uncovered a new SH2 binding site in Bcr-Abl located between Bcr aa242-446, which is different from the Grb2 binding site. Binding could be demonstrated in vitro and also in vivo as shown by co-immunoprecipitation analysis in CML cells. Using a temperature sensitive Bcr-Abl stably overexpressed in hematopoetic cells, we demonstrated that complex formation of Grb10 with Bcr-Abl was kinase activation dependent in vivo. Notably, a Bcr-Abl mutant protein (Bcr/1-242-Abl) which lacks the ability to interact with Grb10 partially alleviated IL-3 dependence of Ba/F3 cells, indicating that the Grb10/Bcr-Abl interaction is important for Bcr-Abl induced IL-3 independence of Ba/F3 cells. In addition, the Bcr/1-242-Abl mutant has a reduced capacity to induce focus formation in fibroblasts. PMID- 9747874 TI - Differential expression and functionally co-operative roles for the retinoblastoma family of proteins in epidermal differentiation. AB - Terminal differentiation requires cell cycle withdrawal, suggesting the involvement of negative cell cycle controllers in the process. We have analysed the involvement of the retinoblastoma family of proteins (pRb, p107 and p130) in epidermal proliferation and differentiation. These proteins play key roles as inhibitors of cell cycle progression and are involved in muscle and neuron differentiation. We found that during in vitro differentiation of human HaCaT keratinocytes, pRb, p107 and p130 are sequentially expressed, in contrast to the co-expression observed during cell cycle progression in the same cells. Immunofluorescence studies on skin sections revealed the presence of pRb and p107 in basal and suprabasal cell layers, whilst p130 is restricted to cells already committed to differentiation in the suprabasal compartments. To explore the functional significance of the differential expression of these proteins, transfection experiments were performed in HaCaT keratinocytes. We observed that the forced over-expression of pRb, p107 or p130 individually did not induce differentiation of the transfected cells. However, the co-transfection of pRb and p107 induced the expression of early differentiation markers (keratin k10) and triple transfectants pRb+p107+p130 expressed markers representative of later stages of epidermal differentiation (involucrin). Finally, we observed that these three proteins repress keratinocyte proliferation, although to a different extent (p107>pRb> or =p130). These results indicate that the members of the pRb family play specific, yet coordinated roles during epidermal differentiation, and that the ordered progression along the different stages of this process results from the effects of different combinations of these proteins. PMID- 9747875 TI - p21Waf1 inhibits the activity of cyclin dependent kinase 2 by preventing its activating phosphorylation. AB - Prostaglandin A2 (PGA2), a potent inhibitor of the growth of many cell types, inhibits G1 phase cyclin dependent kinases (cdk). Although PGA2 suppresses cyclin D1 and elevates p21Waf1 levels, it was the failure of cdk2 to become activated by phosphorylation which correlated best with growth inhibition. In kinetic studies, cdk2 activation was inhibited efficiently only if p21Waf1 levels increased prior to the activating phosphorylation; suggesting that p21Waf1 had blocked this phosphorylation. This model was confirmed in cells from p21Waf1 knockout mice where PGA2 was completely unable to block the activating phosphorylation of cdk2, or inhibit cdk2 activity. As expected, growth inhibition of p21Waf1(-/-) cells was not observed at PGA2 concentrations which inhibited cdk2 activity and growth of p21Waf1(+/+) cells. PMID- 9747876 TI - Molecular features of a new human lymphoma cell line carrying both BCL2 and BCL6 gene rearrangements. AB - Chromosomal translocations and/or their molecular equivalents involving the BCL6 gene on 3q27 band have been suggested to be involved in the development of non Hodgkin's lymphoma of B-cell type (B-NHL). The rearrangement of BCL6 sometimes coexists with other translocations specific to B-NHL. Here, we report a novel B cell lymphoma cell line, YM, established from a patient with diffuse large cell lymphoma. The YM cells expressed B-cell-associated antigens in addition to mu delta/kappa monoclonal immunoglobulin. Southern blot analysis of DNA from YM cells demonstrated rearrangement of the BCL2 gene within the 5' flanking region (5'-BCL2). Polymerase chain reaction (PCR) using primer pairs for the BCL2 exons 1 and 2, and for the constant region of the immunoglobulin kappa light chain gene (IGkappa) revealed PCR products encompassing the 5'-BCL2/IGkappa fusion, indicating that the YM cells had a t(2;18)(p11;q21) translocation. The BCL6 gene was rearranged at a point within the first intron, and cloning of the rearranged BCL6 revealed unidentified sequences juxtaposed to the 5' side of the gene. The isolated clones were mapped to 16p11.2 by high resolution fluorescence in situ chromosomal hybridization. Thus, the YM cells carried a 3q27 translocation involving 16p11.2 as a partner. Chromosome painting of metaphase spreads confirmed that the YM cells had both t(2;18) and t(3;16). Northern blot analysis using a fragment immediately adjacent to the breakpoint on 16p11.2 revealed transcriptional activity within this locus. The YM cells expressed abundant transcripts with aberrant sizes from BCL2 and BCL6, indicating deregulated overexpression of the two genes resulting from the t(2;18) and t(3;16). The YM cell line will therefore be useful to study whether BCL2 and BCL6 genes collaborate in the pathogenesis of B-NHL. PMID- 9747877 TI - Human BAG-1/RAP46 protein is generated as four isoforms by alternative translation initiation and overexpressed in cancer cells. AB - Previously, a Bcl-2-interacting protein, BAG-1, was cloned from mouse cells and was shown to interact with several other proteins and to be important for inhibition of apoptosis. Human BAG-1 (hBAG-1) cDNA, recently isolated by us and two other groups, has been shown to be identical to a hormone receptor-binding protein, RAP46. However, different molecular masses of hBAG-1 protein products were noted by these three groups. Here we demonstrated that hBAG-1 protein was expressed as four isoforms, designated p50, p46, p33 and p29, with apparent molecular masses of 50 kDa, 46 kDa, 33 kDa and 29 kDa, respectively. Deletion, site-directed mutagenesis and in vitro transcription/translation analysis showed that the four protein products of hBAG-1 were expressed by alternative initiation from four different start codons through a leaky scanning mechanism. Furthermore, we demonstrated that the distinct forms of hBAG-1 have different subcellular localizations, suggesting that they may have distinct functions in the cells. Characterization of hBAG-1 RNA and protein also showed that hBAG-1 was overexpressed in human cervical, breast and lung cancer cell lines. Taken together, these data clarify the conflicting observations reported in the literature and suggest that hBAG-1 is expressed as four forms of protein products, which may play a differential role in apoptosis and oncogenesis of human cells. PMID- 9747878 TI - Effector domain mutants of Rho dissociate cytoskeletal changes from nuclear signaling and cellular transformation. AB - The small GTP-binding Rho proteins control a variety of biological activities, including organization of the actin cytoskeleton, regulation of gene expression and cellular transformation. In contrast, Ras proteins do not induce actin stress fibers, but potently transform cells which exhibit a morphology clearly distinct from that caused by activated forms of Rho. To investigate whether nuclear signaling and oncogenic potential of Rho are a consequence of its profound effect on cytoskeletal organization, we replaced each amino acid in the Rho effector loop with those of Ras, or replaced conserved residues with others known to result in differential signaling capability when introduced into Ras and Rac1. These Rho mutants did not gain the ability to induce the MAPK, JNK or p38 pathways but, surprisingly, all Rho effector loop mutants still continued to induce actin stress fiber formation. However, three of these Rho mutants, with substitutions of leucine-39, glutamic acid-39, or cysteine-42, lost the ability to stimulate gene transcription via the serum response factor (SRF) and failed to induce neoplastic transformation. Thus, these results indicate that cytoskeletal changes are not sufficient to induce the transformed phenotype, and that Rho effector molecules regulating the actin cytostructure are distinct from those signaling to the nucleus and subverting normal growth control. PMID- 9747879 TI - Nuclear targeting of Bax during apoptosis in human colorectal cancer cells. AB - Homeostasis in colonic epithelial cells is regulated by the balance between proliferative activity and cell loss by apoptosis. Because epithelial cells at the apex of colonic crypts undergo apoptosis and proliferative activity is usually restricted to the base of the crypts, it has been proposed that the limited availability of growth factor-signals at the upper portions of the crypts may trigger apoptosis. In the present studies, we investigate the mechanism of apoptosis mediated by growth factor deprivation in colorectal carcinoma cells by delineating the possible involvement of Bax and its subcellular localization. We report that inhibition of epidermal growth factor receptor (EGFR) tyrosine kinase activity and downregulation of EGFR by anti-EGFR mAb 225 induces apoptosis in human colorectal carcinoma DiFi and FET cells. Induction of apoptosis was preceded by enhanced expression of newly synthesized Bax protein, and required protein synthesis. In the mAb 225-treated cells, Bax was redistributed from the cytosol to the nucleus and subsequently, to the nuclear membranes. The observed induction of Bax expression by mAb 225 was not associated with p53 induction. However, mAb 225 treatment also triggered relocalization of p53 from the cytosol to a nuclear membrane-bound form. Induction of Bax and its redistribution to the nucleus of DiFi cells during apoptosis was also demonstrated in response to butyrate, a physiological relevant molecule in colonic epithelial cells as it is the principal short-chain fatty acid produced by bacterial fermentation of dietary fiber in colonic epithelium. Using immunofluorescence and confocal microscopy, we observed that Bax is predominantly localized in the cytosol, but during apoptosis it is localized both inside and along the nuclear membrane. Taken together, these findings suggest that apoptosis induced by growth factor deprivation or butyrate may involve the subcellular redistribution of Bax in human colorectal carcinoma cells. PMID- 9747880 TI - Direct association of YY-1 with c-Myc and the E-box binding protein in regulation of glycophorin gene expression. AB - We previously reported that YY-1, a versatile transcription factor, regulates expression of glycophorin gene by binding to its locus control region-like region (Gp-LCR) in combination with E-box binding protein during murine erythroleukemia (MEL) cell differentiation. In the present work, we demonstrated that YY-1 and c Myc, a nuclear oncoprotein, were physically associated in vivo and that down regulation of c-Myc liberated free YY-1 from its complex, resulting in the functional binding of YY-1 to the Gp-LCR. We also showed that the E-box binding protein (EBP) which bound to E-box was physically associated with YY-1, facilitated binding of YY-1 to the neighboring site and their combinatorial binding may stimulate the GpLCR mediated enhancement of erythroid-specific transcription of glycophorin gene in MEL cells. PMID- 9747881 TI - Co-amplification of a novel cyclophilin-like gene (PPIE) with L-myc in small cell lung cancer cell lines. AB - Specific genetic alterations affecting proto-oncogenes of the myc gene family are frequently detected in human lung cancer. Among 11 SCLC cell lines with L-myc gene amplification, four were found to have alteration of the RLF gene by Southern blot and RT-PCR analyses. One cell line, NCI-H378, contained aberrantly sized L-myc-hybridizing bands by Southern and Northern blot hybridization but had no alteration of RLF. Some L-myc-hybridizing cDNAs from NCI-H378 contained a novel sequence with close homology to the cyclophilins joined to antisense L-myc exon 2 sequence. Full length cDNAs isolated from human skeletal muscle containing only the novel sequence identify open reading frames of 301 and 296 amino acids and differ in the C-terminal region by 22 and 17 amino acids. This gene, tentatively named PPIE (peptidyl-prolyl cis-trans isomerase E), has 83% amino acid identity with the central conserved region of cyclophilin A, is evolutionarily conserved by Southern blot, and exhibits differential tissue expression with highest levels found in muscle and brain. Co-amplification of PPIE was observed in seven of eleven L-myc amplified cell lines. Analysis of radiation hybrids suggests that the gene order is RLF-PPIE-L-myc on chromosome 1p and pulse-field gel electrophoresis localizes all three genes to an 800 megabase Mlu I fragment. The prognostic and functional consequences of PPIE gene amplification in SCLC can now be determined. PMID- 9747882 TI - Cyclin D3: requirement for G1/S transition and high abundance in quiescent tissues suggest a dual role in proliferation and differentiation. AB - The mammalian D-type cyclins D1, D2, and D3 activate the cyclin-dependent kinases CDK4 and CDK6 in G1 and thereby promote the cell's commitment to enter S phase. To elucidate the extent of functional overlap among the D-type cyclins, we have examined several aspects of the least characterized member of this subfamily of G cyclin proteins, cyclin D3. Microinjection of cyclin D3-neutralizing antibody inhibited G1/S transition in human (IMR-90) and rat (R12) diploid fibroblasts, indicating that analogous to cyclins D1 and D2, cyclin D3 is essential for timely progression through G1. In contrast to cyclins D1 and D2, cyclin D3 was (i) ubiquitously expressed among a panel of 70 human cultured cell types; (ii) strongly upregulated upon induction of HL-60 leukaemia cells to differentiate; and (iii) accumulated to high levels in a wide range of quiescent cell types in mouse and human differentiated tissues. Complementary analyses of human biopsies and mouse tissues at different stages of foetal and postnatal development revealed lineage-dependent transient or long-term accumulation of the cyclin D3 protein, correlating with initiation/establishment or maintenance of the mature phenotypes, respectively. Our data support the notion that the biological roles of the individual D-type cyclins are not fully redundant, and suggest a possible dual role for cyclin D3 in cell proliferation and induction and/or maintenance of terminal differentiation. PMID- 9747883 TI - In vivo hepatocyte proliferation is inducible through a TNF and IL-6-independent pathway. AB - Recent studies in mice harboring a targeted disruption of genes encoding TNF receptor 1 (TNFR-1) or Interleukin 6 (IL-6) suggested a critical role for TNF and IL-6 in initiation of liver regeneration after 2/3 partial hepatectomy. However, hepatocyte proliferation can also occur following treatment with agents that do not induce tissue loss (primary mitogens). To determine whether the above cytokines could also be involved in mitogen-induced liver cell proliferation, we studied the hepatocyte proliferative response after treatment with primary mitogens in mice knock-out for TNFR-1 or IL-6. Our results showed no difference in the proliferative response of the liver between the wild type and the knock out mice following treatment with the mitogens 1,4-bis[2-(3,5 dichloropyridyloxy)]benzene (TCPOBOP), or the peroxisome proliferator, ciprofibrate, suggesting that TNF or IL-6 may not play a major role in this type of proliferation. Gel shift assay indicated that TCPOBOP-induced hepatocyte proliferation is not associated with activation of STAT3 transcription factor, a major target of IL-6 and other growth factors/cytokines. Our results thus indicate that hepatocyte proliferation can be induced by at least two different pathways; compensatory regeneration being TNF and IL-6-dependent, and mitogen induced direct hyperplasia which does not require TNF or IL-6. PMID- 9747884 TI - DNA damage triggers DRB-resistant phosphorylation of human p53 at the CK2 site. AB - The sequence-specific DNA binding activity of p53 is negatively regulated by a C terminal domain whose phosphorylation in vitro can activate the latent DNA binding function of the protein. The DNA binding activity of p53 is a core component of its stress-activated transcription function, yet it is not yet clear whether phosphorylation within the C-terminal domain plays a role in the p53 damage response in vivo. As the casein kinase 2 (CK2) site at serine 392 is the C terminal phosphorylation motif that exhibits the most pronounced conservation at the primary amino acid level, we have focused on determining whether the CK2 site is modified in vivo and whether radiation effects the extent of that phosphorylation. Using antibodies that can detect serine 392-phosphorylation of p53, we demonstrate that UV radiation can trigger extensive phosphorylation at the CK2 site. The CK2 inhibitor, 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB), can partially inhibit the UV-induced phosphorylation at serine 392, suggesting that CK2 is one of the major serine 392-kinases. However, a striking increase in UV-induced serine 392 phosphorylation and p53 transactivation function at higher levels of DRB suggests that a DRB-resistant/stress-activated pathway may target serine 392 in vivo. These data demonstrate that radiation induced phosphorylation of p53 can occur in vivo at serine 392 and implicate a CK2-independent signal cascade that can function to modulate serine 392 phosphorylation in cells. PMID- 9747885 TI - EMS1 gene expression in primary breast cancer: relationship to cyclin D1 and oestrogen receptor expression and patient survival. AB - The EMS1 and CCND1 genes at chromosome 11q13 are amplified in about 15% of primary breast cancers but appear to confer different phenotypes in ER positive and ER negative tumours. Since there are no published data on EMS1 expression in large series of breast cancers we examined the relationship of EMS1 expression with EMS1 gene copy number and expression of mRNAs for cyclin D1 and ER. In a subset of 129 patients, where matched tumour RNA and DNA was available, EMS1 mRNA overexpression was associated predominantly with gene amplification (P = 0.0061), whereas cyclin D1 mRNA overexpression was not (P = 0.3142). In a more extensive series of 351 breast cancers, there was no correlation between cyclin D1 and EMS1 expression in the EMS1 and cyclin D1 overexpressors (P = 0.3503). Although an association between EMS1 mRNA expression and ER positivity was evident (P = 0.0232), when the samples were divided into quartiles of EMS1 or cyclin D1 mRNA expression, the increase in the proportion of ER positive tumours in the ascending EMS1 mRNA quartiles was not statistically significant (P = 0.0951). In marked contrast there was a significant stepwise increase in ER positivity in ascending quartiles of cyclin D1 mRNA (P = 0.030). A potential explanation for this difference was provided by the observation that in ER positive breast cancer cells oestradiol treatment resulted in increased cyclin D1 gene expression but was without effect on EMS1. The relationship between EMS1 expression and clinical outcome was examined in a subset of 234 patients with median follow-up of 74 months. High EMS1 expression was associated with age > 50 years (P = 0.0001), postmenopausal status (P = 0.0008), lymph node negativity (P = 0.019) and an apparent trend for worse prognosis in the ER negative subgroup. These data demonstrate that overexpression of EMS1 mRNA is largely due to EMS1 gene amplification, is independent of cyclin D1 and ER expression and, in contrast to cyclin D1, is not regulated by oestrogen. Independent overexpression of these genes may confer different phenotypes and disease outcomes in breast cancer as has been inferred from recent studies of EMS1 and CCND1 gene amplification. PMID- 9747886 TI - The Fc receptor (FcR) gamma subunit is essential for IgE-binding activity of cell surface expressed chimeric receptor molecules constructed from human high affinity IgE receptor (Fc epsilon RI) alpha and FcR gamma subunits. AB - The Fc receptor (FcR) gamma subunit was originally discovered as a homodimeric subunit of the high-affinity IgE receptor (FcepsilonRI). But it was recently found to be a common signal-generating subunit of Fc receptors including IgG Fc receptors (FcgammaRs) and IgA Fc receptor (FcalphaR), and furthermore to generate a signal also with stimuli through non-immune receptors. In addition, it plays an essential role in cell-surface expression of the FcepsilonRI and the FcgammaRIIIA isoform and also regulates cell-surface expression and ligand-binding affinity of the FcgammaRI. In this report, we addressed the possibility that the FcRgamma could affect the correct folding of the IgE-binding region of the FcepsilonRIalpha subunit by using the chimeric receptor molecules constructed from human FcepsilonRIalpha and FcRgamma. Furthermore, we demonstrated that the seven amino acid residues in the C-terminal region on the extracellular domain of the FcepsilonRlalpha were essential for maintaining the IgE-binding activity of the FcepsilonRIalpha exodomain on the cell membrane and/or may affect the correct folding of the alpha subunit itself within the cell. PMID- 9747887 TI - The role of T cell receptor dimerization for T cell antagonism and T cell specificity. AB - T cell responses are highly specific and T cell receptors (TCRs) can recognise subtle differences in major histocompatibility complex (MHC)-peptide complexes. While nominal peptide antigens usually act as full agonists that trigger the whole spectrum of T cell responses, some peptides exhibiting mutations at the TCR MHC/peptide contact site stimulate only a fraction of T cell responses (partial agonists) or may even inhibit T cell activation by full agonists (antagonist). The present study analyses mathematically the role of TCR-dimerization for T cell antagonism and T cell specificity in general. It demonstrates that T cell antagonists can effectively inhibit TCR-dimerization and that this mechanism can sufficiently explain all aspects of T cell antagonism. The kinetic model of T cell activation proposes that increasing the time required for effective TCR signaling is the most effective mechanism to increase the discriminatory capacity of TCRs. Our results indicate that TCR-oligomerization is an alternative and efficient mechanism to ensure T cell specificity. PMID- 9747888 TI - T-cell receptor repertoires utilized in response to linear peptides representing an immunodominant MHC class II restricted T-cell epitope are far more diverse than that utilized in response to the same epitope in the nominal antigen. AB - Previous analyses of the T-cell receptor (TCR) repertoire utilized in response to the 1-102 fragment of the lambda cI repressor protein and specific for the immunodominant amino acid 12-26 region in the context of I-Ek, have shown this repertoire to be extremely restricted. In contrast, here we show that the TCR repertoires utilized in two strains of I-Ek expressing mice in response to two linear peptides representing this immunodominant region are diverse. Despite their extensive diversity, these repertoires are somewhat overlapping. In addition, structural similarities were observed between the full lambda cI fragment (1-102) and peptide elicited TCR repertoires, including frequent use of the Valpha2 family of gene segments, particularly among peptide (12-26) elicited TCRs cross-reactive with 1-102/I-Ek. Nevertheless, these data indicate that it may be difficult to mimic the immune response to an immunodominant epitope of a protein antigen via immunization with linear peptides containing the amino acid sequence of that epitope. Possible explanations for differences in the levels of TCR diversity among T cells responding to an epitope present in a nominal antigen as compared to T cells responding to linear peptide antigens containing this same epitope are discussed. PMID- 9747889 TI - Molecular basis of IgE-recognition of Lol p 5, a major allergen of rye-grass pollen. AB - Grass pollen, especially of rye-grass (Lolium perenne). represents an important cause of type I allergy. Identification of IgE-binding (allergenic) epitopes of major grass pollen allergens is essential for understanding the molecular basis of interaction between allergens and human IgE antibodies and therefore facilitates the devising of safer and more effective diagnostic and immunotherapy reagents. The aim of this study was to identify the allergenic epitopes of Lol p 5, a major allergen of rye-grass pollen, immunodissect these epitopes further so that the amino acid residues critical for antibody binding can be determined and investigate the conservation and nature of these epitopes within the context of the natural grass pollen allergens. Peptides, 12-13 amino acid residues long and overlapping each other by 4 amino acid residues, based on the entire deduced amino acid sequence of the coding region of Lol p 5, were synthesised and assayed for IgE-binding. Two strong IgE-binding epitopes (Lol p 5 (49-60) and (265-276), referred to as peptides 7 and 34, respectively) were identified. These epitopes were further resolved by truncated peptides and amino acid replacement studies and the amino acid residues critical for IgE-binding determined (Lol p 5 (49-60) residue Lys57 and (265-276) residue Lys275). Sequences of these epitopes were conserved in related allergens and may form the conserved allergenic domains responsible for the cross-reactivity observed between pollen allergens of taxonomically related grasses. Furthermore, due to its strong IgE-reactivity, synthetic peptide Lol p 5 (265-276) was used to affinity-purify specific IgE antibodies which recognised proteins of other clinically important grass pollens. further indicating presence of allergenic cross-reactivity at the level of allergenic epitope. Moreover, Lol p 5 (265 276) demonstrated a strong capacity to inhibit IgE-binding to natural rye-grass pollen proteins highlighting the antibody accessibility to these sequences within the context of the natural allergens. Strong IgE-binding epitopes of Lol p 5 have been identified down to single critical amino acid residues and are shown to occur as linear or continuous domains in the natural conformation of natural Lol p 5 and other group 5 grass pollen allergens. The fact that such an allergenic synthetic epitope has the capacity to strongly inhibit IgE-binding to natural allergens highlight its potential for use as a candidate in future therapeutics to treat pollen associated allergies. PMID- 9747890 TI - A dominant V beta bias in the CTL response after HSV-1 infection is determined by peptide residues predicted to also interact with the TCR beta-chain CDR3. AB - Many T cell responses are dominated by restricted TCR expression and can range from repeated usage of particular TCR Vbeta- and/or Valpha-elements, to the preferential usage of both V- and J-elements, often in conjunction with conserved V-D-J or V-J junctional sequences. Cytotoxic T lymphocytes specific for a Kb restricted determinant from the herpes simplex virus glycoprotein B (gB) preferentially express a dominant TCRBV10 beta-chain with sequence conservation of a tryptophan-glycine located in the V-D junction. Here we have examined whether immunisation of C57BL/6 mice with the gB-peptide can mimic the CTL response seen after HSV-1 infection. Immunisation with the gB-peptide resulted in the generation of gB-specific CTL that showed a similar TCRBV10 bias to that observed after HSV-1 infection. When the gB-determinant was expressed as a part of a fusion protein, immunised mice again exhibited the TCRBV10 bias with the junctional sequence conservation in the responding CTL. C57BL/6 mice were then immunised with variants of the gB-peptide that contained amino acid substitutions at positions previously predicted to contact the TCR beta-chain CDR3. Analysis of the TCRBV usage of variant specific CTL lines showed that substitutions at the TCR-contact positions 4, 6 and 7 of the gB-peptide resulted in a loss of the TCRBV10 bias. These results suggest that the TCRBV10 bias seen in gB-specific CTL after HSV-1 infection is due to antigenic selection by the minimal peptide and is determined by residues proposed to contact the TCR beta-chain CDR3. PMID- 9747891 TI - Fas has a crucial role in the progression of experimental autoimmune encephalomyelitis. AB - To investigate the role of Fas in experimental autoimmune encephalomyelitis (EAE) in mice, we examined the susceptibility of EAE in C57BL/6 (B6).lpr mice lacking Fas. The frequency of myelin oligodendrocyte glycoprotein (MOG)-induced EAE in B6.lpr mice was significantly lower than that in B6 mice (19% vs 94%). However, no significant difference was observed between them in either the lymphocyte proliferation response or antibody reactivity to MOG. In addition, the histological examination and semiquantitative reverse transcriptase polymerase chain reaction analysis revealed that the infiltration of inflammatory cells and the up-regulation of gene expression for inflammatory cytokines occurred in the central nervous system (CNS) of B6.lpr mice immunized with MOG, even if they showed no clinical sign. These results indicate that Fas may contribute to the pathogenesis of EAE and may play a crucial role in the expansion of inflammation and/or myelin destruction in the CNS rather than in the activation of encephalitogenic T cells in the periphery and/or the breakdown of blood brain barrier. PMID- 9747892 TI - Modulation of innate immune functions by intracerebroventricularly applied neuropeptide Y: dose and time dependent effects. AB - Centrally applied neuropeptide Y (NPY) interacts with the autonomic nervous system and the hypothalamo-pituitary-adrenal (HPA) axis activity. Since these physiological systems have been shown to modulate innate immune functions, the effects of intracerebroventricular (i.c.v.) NPY administration on leukocyte subsets in the blood, spleen and intravascular pool of the lung, blood granulocyte chemiluminescence response, and splenic natural killer (NK) cell mediated lysis were studied in Lewis rats. Concentration-dependent NPY effects were tested at 15 min and 24 h post i.c.v. injection at dosages of 10(-6) M, 10( 9) M, and 10(-12) M. Time dependent effects were investigated at 15 min, 1 h and 24 h after i.c.v. administration of 10(-9) M NPY. Compared to saline controls, an increased number of granulocytes and NK cells in the blood, associated with a decreased granulocyte function and NK cytotoxicity was observed 15 min following NPY infusion. This initial immunosuppression was followed by long lasting stimulatory effects of NPY on the functional capacity of both cell populations when tested at 1 h and 24 h. The dosage of i.c.v. 10(-6) M NPY produced no changes, whilst 10(-9) M produced maximal, and 10(-12) M still significant effects. Results provide evidence that centrally applied NPY influences innate immunity in a dose and time dependent fashion. Cell mobilization from the vascular marginal pool is likely to be an underlying mechanism for the initial immunosuppression. PMID- 9747893 TI - Role of peroxynitrite and poly (ADP-ribosyl) synthetase activation in cardiovascular derangement induced by zymosan in the rat. AB - Peritoneal administration of zymosan in the rat induced a severe inflammatory process characterised by an increase in the plasma levels of nitrite and nitrate, stable metabolites of nitric oxide (NO) and in the levels of peroxynitrite, as measured by the oxidation of the fluorescent dye dihydrorhodamine 123, at 18 hours zymosan challenge. Immunohistochemical examination demonstrated a marked increase in the immunoreactivity to nitrotyrosine, a specific "footprint" of peroxynitrite, in the aorta of zymosan-shocked rats. In ex vivo experiments, thoracic aorta rings of zymosan-treated rats showed a reduced contraction to noradrenaline and reduced responsiveness to the relaxant effect to acetylcholine (vascular hyporeactivity and endothelial dysfunction, respectively). Treatment of zymosan-shocked rats with 3-aminobenzamide or Nicotinamide, inhibitors of poly ADP-ribosil synthetase (PARS) activity reduced the production of peroxynitrite and significantly prevented the cardiovascular dysfunction. Our data suggest that peroxynitrite and PARS activation play a role in the zymosan-induced cardiovascular derangements in the rat. PMID- 9747894 TI - Age-dependent telomere shortening is slowed down by enrichment of intracellular vitamin C via suppression of oxidative stress. AB - Telomeres in eukaryotic somatic cells are destined to the age-dependent shortening, which has not been demonstrated to correlate to direct lesion of telomeric DNA by reactive oxygen intermediates (ROI); still less explicable is the inhibitory effect of ROI-scavenging on telomere shortening. Here, we succeeded in artificial slowdown of age-dependent telomere shortening to 52-62% of the untreated control, in human vascular endothelial cells, by addition of the oxidation-resistant type of ascorbic acid (Asc), Asc-2-O-phosphate (Asc2P), which concurrently achieved both extension of cellular life-span and prevention of cell size enlargement indicative of cellular senescence. The results are attributable to a 3.9-fold more marked enrichment of intracellular Asc (Asc(in)) by addition of Asc2P, subsequently dephosphorylated before or during transmembrane influx, than by addition of Asc itself, and also attributed to diminution of intracellular ROI to 53% of the control level by Asc2P; telomerase activity was at a trace level and underwent an age-dependent decline, which was significantly decelerated by Asc2P. Thus, age-dependent telomere-shortening can be decelerated by suppression of intracellular oxidative stress and/or by telomerase retention, both of which are achieved by enriched Asc(in) but not by extracellular Asc overwhelmingly more abundant than Asc(in). PMID- 9747895 TI - The Hsp90-specific inhibitor, geldanamycin, blocks CD28-mediated activation of human T lymphocytes. AB - The 90 kDa heat shock protein (Hsp90) is a molecular chaperone aiding the folding of nuclear hormone receptors and protein kinases. Hsp90-mediated folding can be disrupted by the Hsp90-specific drug, geldanamycin. Here we provide evidence for the inhibition of the CD28-specific BW 828 antibody-mediated activation of human T lymphocyte proliferation, IL-2 secretion and IL-2 receptor expression by geldanamycin. Our results suggest that the major cytoplasmic chaperone, Hsp90, plays an important role in CD28-mediated T lymphocyte activation. PMID- 9747896 TI - Increase in calcium responsiveness of cardiac myofilament activation in ovariectomized rats. AB - To evaluate a possible role of ovarian sex hormones in the Ca2+ responsiveness of cardiac myofilament activation, the relations of pCa (-log Ca2+ molar concentration) to actomyosin adenosine triphosphatase (ATPase) activity of isolated myofibrillar preparations from 8-10 week ovariectomized (Ovx) rat hearts were compared with those from sham-operated hearts. Deficiency of ovarian sex hormones in plasma of ovariectomized rats was indirectly verified by a significant reduction in uterine weights. Body weights of the ovariectomized rats were significantly greater than those of sham-operated controls. Despite a significant increase in heart weight of 10 week ovariectomized animals, the percent of heart weight-to-body weight ratio was not different from control group. The maximum myofibrillar ATPase activity at pH 7.0 was significantly suppressed after ovariectomy in both eight and ten week groups. However, the maximum ATPase activity at pH 6.5 was significantly suppressed only in 10 week ovariectomized hearts. Surprisingly, in every condition with depressed maximum myofibrillar ATPase activity, the pCa-actomyosin ATPase relationships of ovariectomized cardiac myofilaments demonstrated a significant leftward shift in pCa50 (-log half-maximally Ca2+ activation) from those of sham-operated controls. There was, however, no change in the Hill-coefficient of these cardiac myofilaments after ovariectomy. Analysis of myofilament proteins using gel electrophoresis demonstrated neither change nor loss of any thin filament proteins. These results indicate a possible modulating effect of ovarian sex hormone deficiency on the Ca2+ responsiveness of cardiac myofilament activation by induction of myofilament Ca2+ hypersensitivity but suppression of maximum myofibrillar ATPase activity. PMID- 9747897 TI - Perturbation by lysophosphatidylcholine of membrane permeability in cultured vascular smooth muscle and endothelial cells. AB - Lysophosphatidylcholine (LPC), a major component of oxidized low-density lipoprotein found in atherosclerotic arterial walls, has been shown to have insignificant effect on arterial contraction but cause an impairment of endothelium-dependent relaxation (EDR). The aim of this study was to compare the degree of LPC-induced perturbation in the plasma membrane of cultured aortic smooth muscle cells (SMC) and endothelial cells (EC). In contractility studies phenylephrine (PE) elicited a sustained contraction and a subsequent addition of acetylcholine (ACh) caused an almost complete relaxation. Preincubation of endothelium-intact aortic rings with LPC did not significantly affect PE-elicited contraction but substantially inhibited ACh-triggered relaxation. Such inhibition by LPC was both concentration- and time-dependent. LPC also inhibited relaxation triggered by extracellular ATP and cyclopiazonic acid. Exposure of cultured EC to LPC (30 microM) resulted in an elevation of [Ca2+]i with a lag period of some 25 min. Following [Ca2+]i elevation, addition of Ni2+ resulted in a rapid entry of this ion into the cell. In addition, fura-2 leak-out was observed. Exposure of cultured SMC to 30 microM LPC also resulted in [Ca2+]i elevation and Ni2+ entry. However, LPC did not cause fura-2 leak-out in SMC. Also, LPC raised [Ca2+]i at a slower rate in SMC than in EC. Our results suggest that the plasma membrane of EC is more susceptible to LPC-induced derangement than that of SMC. This may contribute in part to the selective impairment of EDR by LPC. PMID- 9747898 TI - Effects of renal cytoprotective agents on erythrocyte membrane stability. AB - To elucidate potential mechanisms of ischemic renal injury, investigators often use drugs that interfere with specific pathological pathways and study their protective efficacy in in vitro models of ischemia, such as isolated renal proximal tubules subjected to hypoxia. However, the protective effects of certain drugs may depend on non-specific membrane-stabilizing properties. We have studied the effects of several drugs on membrane integrity using osmotic lysis of erythrocytes as a model system. Freshly isolated rabbit erythrocytes were subjected to a hypotonic shock, and the protective effects of various calcium channel blockers, phospholipase inhibitors, free fatty acids, the NO-synthase inhibitor L-NAME, the amino acid glycine and its receptor-analogue strychnine, and two chloride channel blockers were examined. Most agents protected erythrocytes against hypotonic hemolysis when added to the medium in the same concentration range as used in suspensions of hypoxic proximal tubules. Only the protective agents that proposedly act via a blockade of chloride influx (glycine, strychnine and the chloride channel blockers), did not attenuate hypotonic hemolysis. The erythrocyte hemolysis assay may provide an easy and rapid method to screen for non-specific membrane-stabilizing effects of potentially cytoprotective agents. PMID- 9747899 TI - Characterization of antihistamines using biphasic cutaneous reaction in BALB/c mice. AB - Effects of 11 histamine H1 receptor antagonists on IgE-mediated biphasic cutaneous reaction in mice were examined. The immediate phase reaction (IPR) assessed at 1 hour after antigen application was significantly inhibited by all antihistamines examined. The inhibition of IPR by cetirizine and mequitazine were potent, but those by cyproheptadine and diphenhydramine were weak. The later phase reaction (LPR) assessed at 24 hours after antigen application was inhibited by chlorpheniramine, oxatomide, ketotifen, mequitazine, emedastine, terfenadine and azelastine. The inhibition of LPR by emedastine was potent, but those by ketotifen and terfenadine were only partial. Emedastine inhibited both IPR and LPR comparably. Present results indicate that H1 receptor activation is involved in the IPR of the biphasic cutaneous reaction, and that the blockade of H1 receptors at IPR does not contribute to the attenuation of following LPR. Histamine H1 receptor antagonists inhibiting the LPR have a property distinct from H1 receptor antagonism, which may have an additional benefit for the treatment of allergic diseases. PMID- 9747900 TI - A potent nonpeptide neuropeptide Y Y1 receptor antagonist, a benzodiazepine derivative. AB - We describe here a nonpeptide neuropeptide Y Y1 receptor antagonist, 2,4-dioxo 1,5-bis(2-oxo-2-orthotolyl-ethyl)-3-[3-[3-([3-[3-(3-p iperidin-1-ylmethyl phenoxy)-propylcarbamoyl]-propyl]-car bamoyloxymethyl)-phenyl]-ureido]-2,3,4,5 tetrahydro-1H-benzo[b][1,4]diaz epine (Compound 1), which was previously synthesized as a linked type of dual cholecystokinin (CCK)-B and histamine H2 receptor antagonist. Compound 1 competitively inhibited [125I]peptide YY (PYY) binding to Y1 receptors in human neuroblastoma SK-N-MC cells with Ki of 6.4 +/- 1.0 nM, while it had no effect on [125I]PYY binding to Y2 or Y5 receptors even at 1 microM. Functionally, Compound 1 inhibited the Y1 receptor-mediated increase in cytosolic free Ca2+ concentration and Y1 receptor-mediated attenuation of cAMP accumulation in a dose-dependent manner with IC50 values of 95 +/- 5 and 320 +/- 10 nM in SK-N-MC cells, respectively. Neither its CCK-B receptor antagonistic moiety of Compound 1 (Compound 2) nor its histamine H2 receptor antagonistic moiety of Compound 1 (Compound 3) had any effect on [125I]PYY binding, suggesting that the entire structure of Compound 1 is essential for Y1 receptor blocking activity. It showed no significant activity (IC50 > 1 microM) in 30 receptor binding assays and 5 enzyme assays, with the exception of CCK-B and histamine H2 receptors. We conclude that Compound 1 is a useful molecule not only for studying the physiological role of neuropeptide Y but also for exploring more specific Y1 receptor antagonists. PMID- 9747901 TI - Antinociceptive analogs of orphanin FQ/nociceptin(1-11). AB - The presence of pairs of basic amino acids within the sequence of orphanin FQ/nociceptin (OFQ/N) peptide, the endogenous ligand for the ORL1/KOR-3 receptor, has raised the possibility that processing might generate pharmacologically important truncated peptides, including OFQ/N(1-11). OFQ/N(1-11) is pharmacologically active in vivo with a potency comparable to OFQ/N. Several tyrosine-containing analogs of OFQ/N(1-11) have been synthesized and examined for antinociceptive activity. Like OFQ/N(1-11), [Tyr1]OFQ/N(1-11), [Tyr10]OFQ/N(1-11) and [IodoTyr10]OFQ/N(1-11) given supraspinally in mice were antinociceptive in the tailflick assay in mice. The tyrosine analogs showed similar potencies as OFQ/N(1-11) but longer durations of action. This response was readily reversed by the opioid antagonist naloxone despite poor affinities for these analogs at opioid receptors. Another compound, [Tyr11]OFQ/N(1-11) was highly epileptogenic, inducing naloxone-sensitive seizures in greater than 50% of the mice tested at doses comparable to those examined with the other analogs. These results indicate that it is possible to make analgesic OFQ/N(1-11) analogs. The activity of [IodoTyr10]OFQ/N(1-11) suggests that it may prove useful as a radioligand in exploring potential OFQ/N(1-11) binding sites. PMID- 9747902 TI - Naltrexone, but not atropine or yohimbine, antagonizes suppression of formalin induced spinal sensitization by intrathecal nociceptin. AB - We investigated the effects of spinal nociceptin on formalin-induced spinal sensitization and examined the role of the opioidergic, alpha 2-adrenergic and muscarinic cholinergic receptors in the nociceptin-produced suppression of spinal sensitization. The results demonstrated that spinal nociceptin suppressed the formalin-induced spinal sensitization in a dose-dependent manner (1, 5 and 10 nmol). The inhibitory effect of 10 nmol of nociceptin on spinal sensitization, was readily antagonized by naltrexone, but not by atropine or yohimbine. Each of the antagonists, naltrexone, atropine or yohimbine, alone had no effect on the formalin-induced spinal sensitization. Our results show that spinal nociceptin elicits dose-dependent, naltrexone-reversible suppression of spinal sensitization evoked by injection of formalin. PMID- 9747903 TI - I1-imidazoline receptors: an update. AB - BACKGROUND: The site of the hypotensive action of imidazoline compounds, such as clonidine, was first identified within the nucleus reticularis lateralis of the rostroventrolateral part of the medulla (NRL/RVLM). It was shown that imidazolines and related substances reduced blood pressure when applied in this area whereas no catecholamine was capable of such an effect. IMIDAZOLINE-SPECIFIC BINDING: We previously suggested the existence of receptors specific for imidazoline-like compounds that differed from the alpha-adrenergic receptors. Imidazoline-binding sites were subsequently reported in the brain and in a variety of peripheral tissues, including the human kidney, and as expected these specific binding sites do not bind the catecholamines. The imidazoline-binding sites are classified into two subgroups: the I1-type, which is sensitive to clonidine and idazoxan, and the I2-type, sensitive to idazoxan and largely insensitive to clonidine. Numerous studies have confirmed the involvement of these receptors in various regulations and pathological processes, hypertension being the most notable. DRUGS: Functional studies have confirmed that the hypotensive effects of clonidine-like drugs involve I1-imidazoline receptors while their most frequent side effects only involve alpha2-adrenergic receptors. Recent studies have shown that a contribution of both receptor types might be necessary to trigger the hypotensive effect of central origin. Rilmenidine, an oxazoline analogue to the imidazolines, has been proposed as the prototype of a new class of antihypertensive drugs selective for I1-imidazoline receptors. At hypotensive doses, this drug is devoid of any significant sedative effect. As with clonidine, it evokes hypotension when injected into the NRL region and it completely displaces the [3H]clonidine bound to specific imidazoline-binding sites in human medullary membrane preparations, but it has proved more selective for cerebral imidazoline receptors than clonidine. This selectivity might explain the low incidence of side effects evoked by rilmenidine. CONCLUSION: Rilmenidine is the first example of a drug exhibiting a favourable selectivity between I1 imidazoline receptors and alpha2-adrenergic receptors, for example reducing blood pressure but avoiding sedation and mouth dryness. PMID- 9747904 TI - Site and receptors involved in the sympathoinhibitory actions of rilmenidine. AB - INTRODUCTION: It is well accepted that centrally acting antihypertensive agents such as rilmenidine reduce blood pressure through inhibition of the sympathetic nervous system. The central receptor involved, be it a central imidazoline receptor or an alpha2-adrenoceptor and its location, is less certain. METHODS AND RESULTS: The present paper reviews studies from our laboratory examining these questions by using antagonists with differing affinity for imidazoline receptors and alpha2-adrenoceptors. In addition, we have used various routes of administration in conscious and anaesthetized normotensive rabbits. We found that rilmenidine was more potent in its hypotensive action when administered into the fourth ventricle than when given intravenously and considerably more potent when injected into the rostroventrolateral medulla (RVLM) compared to the nucleus of the solitary tract (NTS). By contrast, alpha-methylnoradrenaline, which acts solely through alpha2-adrenoceptors to produce hypotension, was similarly potent in both the NTS and RVLM. Injections of rilmenidine into the RVLM reduced renal sympathetic tone and produced a marked inhibition of renal sympathetic baroreflex responses. The pattern of renal sympathetic baroreflex effects of rilmenidine administered into the RVLM was similar to the effects of the fourth ventricular or intravenous administration. These studies together support the view that the RVLM is the major site of action. We have determined the relative importance of imidazoline receptors and alpha2-adrenoceptors in the inhibition of renal sympathetic nerve activity produced by rilmenidine administered into the RVLM, the fourth ventricle or intravenously. Initial studies in conscious rabbits showed that intravenous or fourth ventricular administration of rilmenidine induced renal sympatho-inhibition which was preferentially reversed by idazoxan or efaroxan (imidazoline/alpha2-adrenoceptor antagonist) compared to 2 methoxyidazoxan (alpha2-adrenoceptor antagonist). In anaesthetized rabbits, administration of idazoxan into the RVLM reversed the inhibition of the renal sympathetic activity induced by RVLM or intravenous administration of rilmenidine. In contrast, idazoxan had no effect on the responses produced by the alpha2-adrenoceptor agonist alpha-methylnoradrenaline. CONCLUSION: Our studies suggest that rilmenidine given systemically acts primarily on imidazoline receptors in the RVLM to reduce sympathetic tone and to modulate sympathetic baroreflexes. PMID- 9747905 TI - Complementary antihypertensive action of rilmenidine on the pressure-natriuresis relationship and sodium preference in spontaneously hypertensive rats. AB - Previously, changes in position and slope of the pressure-natriuresis relationship have been used to characterize antihypertensive drugs in basic research. Rilmenidine may chronically reduce arterial pressure via central nervous system and renal imidazoline receptors. The present experiments were used to examine the shift in the pressure-natriuresis relationship during rilmenidine administration. We examined the effects of twice daily doses (1 and 3 mg/kg) for 6 days on the pressure-natriuresis relationship determined for control and treated spontaneously hypertensive rats (SHR) drinking tap water or 1% NaCl. The pressure-natriuresis relationship was shifted to the left for the 3 mg/kg dose and the slope was no different from the control. These experiments also indicated that rilmenidine might have an effect on sodium preference which was confirmed in a third series of experiments by permitting control and treated (3 mg/kg) SHR access to both tap water and 1% NaCl. This lack of change in slope indicates that, during rilmenidine treatment, the arterial pressure is relatively insensitive to sodium intake. The shift to the left indicates a restoration of the pressure-natriuresis relationship after chronic treatment with rilmenidine and a resetting of the long-term blood pressure control. Rilmenidine also reduces salt appetite in the SHR. PMID- 9747906 TI - High blood pressure management: potential benefits of I1 agents. AB - SYMPATHETIC NERVOUS SYSTEM AND HYPERTENSION: Biochemical, electrophysiological, pharmacological and haemodynamic findings support the existence of sympathetic nervous system activation in primary human hypertension. Analysis of regional sympathetic nervous system function, using both neurophysiological methods for measuring sympathetic nerve firing rates, and neurochemical techniques for quantifying regional noradrenaline spillover to plasma has demonstrated activation of the sympathetic nervous outflows to the heart, the kidneys, and skeletal muscle vasculature, particularly in younger patients. The initiating cause of this sympathetic nervous stimulation is unknown, but estimation of central nervous system noradrenaline turnover in hypertensive patients, using measurements of the washout of noradrenaline and its lipophilic metabolites into the internal jugular veins, indicates that activation of forebrain pressor noradrenergic nuclei is the probable underlying mechanism. CONSEQUENCES OF INCREASED SYMPATHETIC ACTIVITY: The sympathetic activation present in human hypertension no doubt contributes to the blood pressure elevation, and is a legitimate target for therapeutic intervention with imidazoline receptor-binding agents such as rilmenidine. In addition, the sympathetic nervous activation seems to have adverse consequences in hypertensive patients beyond initiating the blood pressure elevation. There is evidence that neural vasoconstriction has metabolic effects, in skeletal muscle impairing glucose delivery to muscle, causing insulin resistance and hyperinsulinaemia, and in liver retarding postprandial clearing of lipids, contributing to hyperlipidaemia. Cardiac sympathetic activation is demonstrably a cause of sudden death in heart failure patients; a comparable arrhythmogenic effect is probable in hypertension. A trophic effect of sympathetic activation on cardiovascular growth is also likely, contributing to the development of left ventricular hypertrophy. Rilmenidine, through its central nervous system actions, has been demonstrated to powerfully reduce sympathetic nervous activity in essential hypertension patients. INHIBITING THE SYMPATHETIC SYSTEM: As the clinical consequences of sympathetic nervous activation in essential hypertension appear to go beyond that of hypertension pathogenesis, extending to a causal influence in atherosclerosis development, cardiovascular hypertrophy and cardiac arrhythmias, it is possible that, of all antihypertensive drugs, those inhibiting the sympathetic nervous system might best reduce cardiovascular risk. This remains to be tested. PMID- 9747907 TI - Clinical relevance blood pressure variability. AB - Blood pressure fluctuates continuously over time, either spontaneously or in response to a variety of external stimulations. The occurrence of these continuous and often marked blood pressure variations is not only of pathophysiologic interest, but it may also have a clinical relevance. Indeed, it has been shown that the occurrence of pronounced blood pressure changes at the time of the physician's visit may introduce errors in the diagnosis of hypertension and in the assessment of the efficacy of antihypertensive treatment. Moreover, several studies have reported that the end-organ damage of hypertension is significantly and independently related to the degree of blood pressure variability during the day and night. This was shown by reports that assessed blood pressure variability by a variety of different methods, i.e. by computing the 24 h or daytime blood pressure standard deviation, the degree of morning blood pressure rise or that of night-time blood pressure fall, the frequency of blood pressure peaks over the 24 h, and the blood pressure increases under stressful conditions or during physical exercise. Results from a recent follow-up study have provided evidence that the degree of blood pressure variability may also have prognostic relevance in hypertensive patients. Thus, optimal antihypertensive treatment might also need to reduce the degree of blood pressure fluctuations together with the 24 h average blood pressure levels. Until recently, however, available antihypertensive drugs have been ineffective in buffering blood pressure variability or have even been responsible for an increase in the degree of blood pressure fluctuations. Further studies are needed to assess whether recently developed antihypertensive agents, and in particular those able to induce a smooth reduction in blood pressure over the 24 h or to modulate the sympathetic influences exerted on the cardiovascular system, may represent better tools to reduce the magnitude of an enhanced blood pressure variability in hypertensive patients over the 24 h. Recent progress in technology has offered us more powerful tools to address this issue. They include devices for continuous noninvasive ambulatory blood pressure monitoring (Portapres, TNO), and techniques for a more comprehensive analysis of all components which contribute to overall blood pressure variability (broad-band spectral analysis). PMID- 9747908 TI - Noradrenaline spillover and microneurography measurements in patients with primary hypertension. AB - Quantification of sympathetic nervous activity is important in hypertension as the sympathetic system may play both long-term and short-term roles in cardiovascular regulation; it may be involved in the initiation of hypertension, the complications of hypertension, and anti-adrenergic drugs also reduce mortality. Of the clinical tests available at present, microneurography and noradrenaline overflow are measures of different aspects of sympathetic activity. Other indirect measures including reflex testing, autonomic blockade, and power spectral analysis provide indices of sympathetically mediated responses. The latter are therefore influenced by variations in function not only of the sympathetic system but also the organ mediating the response under investigation. Plasma noradrenaline concentration measurements are confounded by the influence of noradrenaline plasma clearance on plasma concentration. Antecubital venous samples represent the venous drainage of the forearm and are not typical of other vascular beds. The inability to detect regional differentiation of sympathetic responses is a feature of all 'global' measures of sympathetic function. Microneurographic recordings of sympathetic nerve fibre firing rates provide direct and continuous measurements of efferent activity to the skin and muscle. Regional noradrenaline spillover measurements, performed with infusions of radiolabelled noradrenaline and sampling from centrally placed catheters, may be used clinically to study internal organs not accessible to nerve recording with microneurography. Primary hypertension is characterized by preferential activation of the cardiac, skeletal muscle and renal sympathetic outflow particularly in younger patients. Central nervous system overflow of noradrenaline is also increased, providing an attractive rationale for the investigation of new centrally acting antihypertensive drugs. Activation of forebrain pressor noradrenergic nuclei may be an underlying mechanism for sympathetic activation in a proportion of patients with primary hypertension. PMID- 9747909 TI - Preventive effect of rilmenidine on the occurrence of neurogenic ventricular arrhythmias in rabbits. AB - BACKGROUND: Centrally acting antihypertensive drugs bearing an imidazoline or a related chemical structure inhibit sympathetic nervous output to the heart and vascular beds, and enhance parasympathetic tone. Cardiac ischaemia and ventricular arrhythmias that can result from hypertension are likely to benefit from such effects. OBJECTIVE: To investigate the effects of rilmenidine, an oxazoline with antihypertensive properties, in a model of neurogenically induced ischaemic ventricular arrhythmias. METHODS AND RESULTS: Bicuculline, a alpha aminobutyric acid (GABA(A)) receptor antagonist, was administered intracisternally in pentobarbitone anaesthetized rabbits; 10 microg/kg intracisternal bicuculline induced polymorphic ventricular ectopic beats and ventricular tachycardia, while blood pressure increased by about 50-60% and heart rate in sinus rhythm decreased by about 20%. Rilmenidine pretreatment (10 min), either administered intravenously (0.01, 0.1, 1 mg/kg) or intracisternally (3, 10, 30 microg/kg), dose-dependently prevented the occurrence of bicuculline induced arrhythmias and, because of a lower baseline, the blood pressure values reached were less when compared with controls. Intracisternal idazoxan (15 microg/kg) had no significant antiarrhythmic effect but antagonized, in part, the haemodynamic and antiarrhythmic effects of rilmenidine (1 mg/kg intravenously; 30 microg/kg intracisternally). CONCLUSION: The antiarrhythmic effects observed with rilmenidine are mainly mediated by blunting the bicuculline-induced increase in the sympathetic nervous output to the heart and the vascular beds. These effects of rilmenidine are likely to originate from action on the central as well as on the peripheral nervous systems. Direct coronary or cardiac effects might also play a role, in particular at low non-hypotensive intravenous doses. PMID- 9747910 TI - Rilmenidine normalizes fructose-induced insulin resistance and hypertension in rats. AB - OBJECTIVE: The aim of this study was to determine the effects of rilmenidine (an antihypertensive drug that lowers blood pressure by decreasing sympathetic outflow) in an animal model of hypertension associated with insulin resistance, i.e. rats fed on a high-fructose diet. DESIGN: Wistar rats were fed for 4 weeks either on a standard diet (S group) or on a high-fructose diet (F group; 34.5% fructose). In half of the rats in the F group, rilmenidine (1 mg/kg per day) was added to the drinking water for the last 2 weeks of the diet (FR group). RESULTS: Body weight gain was higher in the F than in the S rats (66+/-8g versus 45+/-8g, P< 0.05), but was prevented by rilmenidine treatment (32+/-2g). Arterial systolic blood pressure was increased in F rats (162+/-2 versus 155+/-2 mmHg, P< 0.05), rilmenidine reduced this value to normal (149+/-3 mmHg). Glucose tolerance, glucose turnover rate, and insulin secretion were not modified by the diet or by the drug. However, during a euglycemic hyperinsulinemic clamp, glucose utilization was lower (10+/-1 versus 14+/-1.5 mg/min per kg; P< 0.05) and hepatic glucose production higher (1+/-0.01 versus 0 mg/min per kg, P< 0.01) in F than in S rats. These changes in insulin action were totally abolished by rilmenidine. CONCLUSIONS: These data demonstrate that rilmenidine can ameliorate the deleterious effects of a high-fructose diet, i.e. weight gain, hypertension, and resistance to the effects of insulin. PMID- 9747911 TI - Sympatho-adrenal mechanisms regulating cardiovascular hypertrophy in primary hypertension: a role for rilmenidine? AB - OBJECTIVE: Overactivity of the sympatho-adrenal system has long been considered a major factor contributing to blood pressure elevation in primary hypertension in humans and experimental animals. Our aim has been to elucidate its role in the development of cardiovascular hypertrophy in hypertensives. METHODS: Two studies have been performed in spontaneously hypertensive rats (SHR). One involved irreversible inhibition of the sympatho-adrenal system in newborn SHR using a sympathectomy procedure combined with prolonged alpha1-adrenoceptor blockade. The other involved reversible, long-term inhibition of the sympatho-adrenal system in young but mature SHR, by treatment with rilmenidine, a centrally active antihypertensive agent interacting with imidazoline receptors. Their effects on cardiovascular structure were examined. RESULTS: Sympathectomy plus alpha1 adrenoceptor blockade prevented the development of cardiac and vascular hypertrophy in adolescent SHR and these effects were maintained later in life. Rilmenidine administered to older (9-week) SHR also attenuated cardiac hypertrophy, abolished perivascular fibrosis associated with the intramyocardial vessels, and normalized vessel structure in the richly sympathetically innervated mesenteric vasculature. These effects were only partially related to the level of blood pressure reduction. CONCLUSIONS: Inhibition of the sympatho-adrenal system not only reduces blood pressure to normotensive levels in SHR but also has beneficial effects on cardiovascular structure, potentially reducing risk factors for cardiac and renal abnormalities frequently seen in long-term hypertensives. Therapeutically, these effects are likely to be achieved with rilmenidine. PMID- 9747912 TI - Regression of left ventricular hypertrophy in hypertensive patients after 1 year of treatment with rilmenidine: a double-blind, randomized, controlled (versus nifedipine) study. AB - OBJECTIVE: To assess the effect of 1-year treatment with rilmenidine, an oxazoline compound that exerts its antihypertensive effects through binding to imidazoline receptors in the brainstem, on left ventricular hypertrophy (LVH) secondary to essential, mild-to-moderate hypertension [supine diastolic blood pressure (DBP)95-115 mmHg]. METHODS: We performed a double-blind, randomized, controlled (versus slow-release nifedipine) trial. Adjustment of treatment took place every month (M) between inclusion (MO) and an evaluation after 6 months (M6), then during M9 and after 1 year (M12) to achieve supine DBP values < or = 90 mmHg. Patients were dropped from our study if they had DBP> 95mmHg during two consecutive visits or DBP>115 mmHg on one occasion. The daily dosage of rilmenidine was 1 mg, and could be increased to 2 mg/day. The daily dosage of slow-release nifedipine was started from the beginning at the maximum dosage of 40 mg/day, so that there was no true adjustment of treatment despite the allocation of patients to a different unit in the case of DBP> 95 mmHg. The primary criterion was the change in left ventricular mass index (LVMI, g/m2), assessed by echocardiography, between MO and M12 for patients who completed the trial. RESULTS: After a 1-month placebo run-in period, 76 patients were selected and 73 were included (35 treated with rilmenidine and 38 treated with nifedipine). Fifteen patients withdrew from the study and two completed the study with a major deviation from protocol, leaving 56 patients (24 treated with rilmenidine and 32 treated with nifedipine) for a per-protocol analysis. Baseline demographic characteristics and history of arterial hypertension for the rilmenidine and nifedipine groups were similar, for included patients and for those taken into account for the per-protocol analysis. Between MO and M12, DBP in members of the per-protocol population was adequately controlled for those in the rilmenidine group (102.7+/-4.6 versus 88.5+/-7.1 mmHg, respectively) and for those in the nifedipine group (102.7+/-5.1 versus 85.6+/-79 mmHg, respectively). During MO, LVMI of patients in the rilmenidine group (176.9+/-41.3 g/m2) was slightly higher than that of patients in the nifedipine group (172.6+/-35.1 g/m2). During M12, LVMI was observed to have decreased both for patients in the rilmenidine group (to 154.8+/-40.2 g/m2, a decrease of 22.1+/-23.3 g/m2, P< 0.001) and for those in the nifedipine group (to 145.6+/-36.4 g/m2, a decrease of 26.9+/-29.5 g/m2, P< 0.001) but the difference between these two groups was not significant (P= 0.5). CONCLUSION: One-year treatment with a daily dosage of 1 or 2 mg rilmenidine achieves a significant reduction of left ventricular mass, which is not statistically different than that occurring with a daily dosage of 40 mg of slow-release nifedipine. PMID- 9747913 TI - Unstable mutations and neurodegenerative disorders. AB - Trinucleotide repeat expansions are involved in an increasing number of neurodegenerative disorders. Eight disorders are caused by translated CAG expansions with sizes usually below 100-200 repeats. Expansions are observed in unrelated genes, and the threshold above which the disease becomes manifest varies according to the locus. There is a strong negative correlation between age at onset and the number of repeats. Direct molecular diagnosis, which is now possible, allows classification according to genotype, thereby multiplying the number of related disorders. Molecular analysis is also useful to diagnose disorders with variable and overlapping clinical features. Recent findings suggest that intranuclear inclusions are a characteristic of disorders with translated CAG expansions. Their formation might constitute an important step in the pathological process. Friedreich ataxia is the first disorder caused by a trinucleotide repeat expansion located within an intron. The clinical spectrum of the disease and its diagnostic criteria have been recently reevaluated in a large series of patients. Interestingly, Friedreich's ataxia is now thought to be associated with intramitochondrial iron accumulation. Frataxin, the protein that is mutated, might normally be responsible for mitochondrial iron homeostasis in tissues that are affected by the disease. PMID- 9747914 TI - Secondary dystonias. AB - Secondary or symptomatic dystonias are (1) often accompanied by other neurological deficits. (2) begin suddenly at rest and occur at rest from the onset, (3) are associated with different hereditary and environmental causes. From an aetiological point of view, secondary dystonias can be caused by focal brain lesions of various origin, neurodegenerative disorders, metabolic disorders of the central nervous system (CNS), and several drugs and chemicals that affect the basal ganglia, thalamus and brain stem. Furthermore, secondary (focal) dystonias can be caused by peripheral injury. In the following review, we will discuss epidemiology, genetics, pathogenesis, neuroimaging, neuropathology, clinical manifestation, clinical course and differential diagnosis of secondary dystonias. Therapeutic options are given depending on the aetiology and the topological type of dystonia. PMID- 9747915 TI - Clinical correlates of granulomas in muscle. AB - We evaluated the clinical and myopathological features of all patients with granulomas in muscle biopsy specimens identified over a 5-year period (1992-1996) at the Washington University Medical Center. Ten patients were found to have granulomas in their muscle biopsy specimens. Of these, eight patients had myopathic changes. Seven had dysphagia as a major functional difficulty during the course of their disease. None had elevated levels of serum creatine kinase (CK). Four of the patients with myopathy had systemic sarcoidosis and relatively severe proximal weakness with functional disability. Treatment with corticosteroids was followed by marked improvement in strength and functional disability. The four other patients with myopathy had no systemic signs of sarcoidosis. Weakness was especially prominent distally in three of these patients. The two patients in this group treated with corticosteroids did not improve. The final two patients, who had granulomas in muscle but no myopathic changes, had clinical syndromes of mononeuritis multiplex and eosinophilic fasciitis (Shulman syndrome). We conclude that granulomatous myopathy, in the presence or absence of systemic sarcoidosis, is commonly associated with dysphagia (87%) and a normal serum CK. Clinical features in patients with sarcoidosis included severe proximal weakness with functional disability that often responded to corticosteroid treatment. Granulomatous myopathy without systemic sarcoidosis was associated with milder, but more predominantly distal weakness. PMID- 9747916 TI - The role of morpho-volumetric and memory correlations in the diagnosis of early Alzheimer dementia. AB - The aim of the present study was to assess selective atrophy of the temporal lobe and amygdala in the early stages of Alzheimer dementia (AD). Magnetic resonance imaging (MRI) measurements and the presence of highsignal lesions (HSL) were analysed in 31 patients with mild to moderate probable AD and 22 controls. In the AD group, MRI findings were compared with cognitive variables and specific features of memory functions. Alzheimer patients showed a significant reduction in volumetric measurement compared with controls in the total volume (P < 0.01), temporal lobe (P < 0.01) and amygdala (P < 0.05). The temporal lobe/brain volume ratio was also significantly reduced in AD subjects (P < 0.05). Atrophy of temporal structures was significantly related to the degree of episodic and semantic memory impairment according to a material-specific effect. No significant correlations between amygdala and cognitive variables were found. The results of our study confirm the usefulness of measures of temporal lobe atrophy assessed with MRI in the diagnosis of AD. In contrast, HSL are relatively common in AD patients (12/31 cases) and were not related to volumetric findings, severity of dementia or functional disability. PMID- 9747917 TI - Somatosensory evoked potential studies in internal capsule and corona radiata infarction. AB - To document the somatosensory evoked potential (SEP) changes in capsular and corona radiata infarction and correlate these with clinical and radiological findings, 15 patients with corona radiata and 16 with internal capsular infarction were studied. The mean age of the patients was 55 years (range 26-80), and 6 of them were female. In the patients with corona radiata infarction, median N9-N20 conduction time was abnormal in 4 cases, which correlated with sensory abnormalities in 1. In 3 of these patients, infarction was located in the anterior two-thirds and in 1 there was total corona radiata infarction. The amplitude of N20 potential on the affected side was reduced in 1 patient. In the capsular infarction group, N9-N20 conduction time was abnormal in 1 patient only who had total involvement of the posterior limb of the internal capsule. The amplitude of N20 was reduced in another patient. There were 4 patients who had abnormal sensory findings, but their SEPs were normal. At 3 months, the SEP changes remained stable in all of the patients who were followed up. The SEP changes did not correlate with changes in sensation or 3-month outcome as assessed by the Barthel index score. The lack of clinicoradiological and SEP correlation may be owing to variation on the organisation of sensory pathways in the corona radiata and internal capsule. PMID- 9747918 TI - Electrophysiological brain stem investigations in idiopathic narcolepsy. AB - Narcolepsy is associated with various rapid eye movement (REM) sleep abnormalities. Distinct brain stem areas seem to play a prominent role in REM sleep regulation. Recent magnetic resonance imaging (MRI) studies have led to conflicting findings concerning the presence of structural brain stem lesions in patients with idiopathic narcoleptic syndrome. However, multimodal electrophysiological brain stem investigations may reveal functional brain stem abnormalities even in the absence of MRI abnormality. Therefore we investigated brain stem function in 12 idiopathic narcoleptic patients by systematically studying tegmental brain stem pathways. All of the patients met the diagnostic criteria of the International Classification of Sleep Disorders, with typical changes in polysomnography and the multiple sleep latency test. Electrophysiological investigations comprised masseter reflex, blink reflex, masseter inhibitory reflex, early auditory evoked potentials and electrooculography with vestibular testing. In no patient were electrophysiological brain stem abnormalities observed. Our findings do not support the existence of a relevant brain stem lesion in narcoleptic patients with normal neurological status. PMID- 9747919 TI - The effect of oral ethanol consumption on eye movements in healthy volunteers. AB - Horizontal and vertical eye movements were recorded and analysed with an infrared photoelectric technique in 12 healthy volunteers under various blood alcohol concentrations (0.0, 0.5, 1.0 g/kg body weight, [% per thousand]). The predictive smooth-pursuit tracking and saccadic eye movements were studied in response to unpredictable target jumps and during scanning of a classical kitchen scene and a traffic scene. Smooth-pursuit eye movement gain value decreased dose-dependently and was compensated by an increased number of catch-up saccades. With increasing blood alcohol concentrations peak velocities of horizontal and vertical visually guided reflexive saccades decreased while their latencies to the target increased. At blood alcohol concentrations of 0.5% per thousand and 1.0% per thousand healthy volunteers showed significantly longer mean fixation durations and a lower total number of exploratory saccades when scanning both the classical kitchen scene and the traffic scene. Surprisingly, in both of these scanning tasks the total fixation duration or the relative number of exploratory saccades increased in those scene sectors in which exciting situations were presented. Additionally, the time interval needed to foveate these exciting areas for the first time increased, probably due to an attention deficit. In conclusion, these findings indicate that alcohol consumption impairs the velocity and initiation of saccadic and smooth-pursuit eye movements, but that subjects can nevertheless still recognize exciting and relevant areas of visual scenes. The significant increase in fixation time, however, does not allow scanning of the entire visual scene during an adequate period of time. Therefore the reduced visual exploration caused by alcohol reflects an impaired sensorimotor processing of active visual perception. PMID- 9747920 TI - Paralytic shellfish poisoning: clinical and electrophysiological observations. AB - In paralytic shellfish poisoning a mollusc contaminated with a toxin (saxitoxin) causes a potentially lethal disease, clinically characterised by gastrointestinal and neurological symptoms, of which possible respiratory depression is the most serious. The toxin acts by blocking the sodium channels. We report 9 Portuguese patients with this disease. The mollusc was identified as Mytilus edulis, contaminated with the dinoflagellate Gymnodinium catenatum, and the toxin saxitoxin. Our patients had a benign clinical course with cerebellar ataxia as the most severe neurological impairment. Eight out of 9 patients had neurophysiological investigations, the largest number so far reported. Motor and sensory conduction velocities and amplitudes were normal. The proximal conduction times, as assessed by F waves, showed delayed conduction and decreased frequency, which returned to normal in few weeks. The somatosensory evoked potentials confirmed normal peripheral and central sensory conduction. The rich vascular supply at root level of the sodium channels of the proximal motor nerves may explain the greater vulnerability to toxin damage. The typically transient and quickly reversible nerve dysfunction caused by ion channel blockade is reported. PMID- 9747921 TI - Skeletal muscle disturbances may precede clinical and laboratory evidence of autoimmune hypothyroidism. PMID- 9747922 TI - Progressive multifocal leukoencephalopathy presenting as a solitary gray matter lesion. PMID- 9747923 TI - Opsoclonus following Epstein-Barr virus infection. PMID- 9747924 TI - Juvenile distal spinal muscular atrophy: a case with Arnold-Chiari malformation. PMID- 9747925 TI - Medical Editors' Trial Amnesty (META) PMID- 9747926 TI - Amyotrophic lateral sclerosis: an introduction. AB - Amyotrophic lateral sclerosis (ALS) is a progressive, neurodegenerative disease that results in the degeneration of lower and upper motor neurones in the brain and the spinal cord. Early onset and modern therapies, including assisted ventilation, improve survival in this disease, although its cause remains uncertain. Amongst the possible causes are deficiency of nerve growth factor, deficient glutamate re-uptake, autoimmunity and mutation of superoxide dismutase 1 gene. Additional factors may be industrial pollutants and occupational exposure to chemicals associated with welding and soldering. The criteria for the diagnosis of ALS, proposed by the World Federation of Neurology, are presented together with a review of the clinical features of the disease. PMID- 9747927 TI - Ethical considerations in disease management of amyotrophic lateral sclerosis: a cross-cultural, worldwide perspective. AB - Amyotrophic lateral sclerosis (ALS) is universally fatal. Technological advances have provided a means to impact upon, without radically improving, the natural history of the disease. In addition, we now have the capability of potentially identifying patients who are pre-symptomatic carriers of the rare heritable forms of the disease. These capabilities provide the basis for the numerous ethical dilemmas that face patients, physicians, and agencies responsible for health care expenditures; dilemmas that can only be amplified between cultures. This paper attempts to address some of the major ethical issues germane to the care of ALS patients. It discusses the emergence of autonomy as the reigning principle of medical ethics in the United States and its potential conflict with the ethical dilemma of limited resource allocation. Finally, it attempts to compare and contrast, in an admittedly anecdotal and fragmentary fashion, the perspective of other cultures regarding the care of ALS patients. PMID- 9747928 TI - Mechanical ventilation in amyotrophic lateral sclerosis: a cross-cultural perspective. AB - Mechanical ventilation is known to be an effective means of relieving symptoms of chronic hypoventilation and prolonging life in patients with amyotrophic lateral sclerosis (ALS). Various methods of mechanical ventilation are available to patients with ALS. However, attitudes towards mechanical ventilation in ALS vary widely across different cultures, and even within a given medical system. This article describes differences and similarities between a North American, a European and a Japanese approach, based on the respective medical and cultural traditions. The common goal is to provide optimal palliative care to patients with ALS. PMID- 9747929 TI - Nutritional management in amyotrophic lateral sclerosis: a worldwide perspective. AB - Although respiratory failure is the primary cause of death in patients with amyotrophic lateral sclerosis (ALS), the management of nutritional status is important to enhancing the quality of life and optimising the timing of interventive techniques. Progressively weakening muscles impair the patient's ability to eat, and nearly all patients with ALS develop severe dysphagia. If nutritional support is not provided, food and fluid consumption may be greatly restricted, leading to weight loss and malnutrition. This may be compounded by impaired respiratory functions, which place increased energy demands on the patient. This paper describes the nutritional needs of ALS patients from a worldwide and cross-cultural perspective. In particular, the differences between a paternalistic and a patient-centred approach to treatment are addressed. The need for further study into the nutritional status of ALS patients and the issue of parenteral and enteral nutritional therapy, particularly percutaneous endoscopic gastrostomy, are discussed. PMID- 9747930 TI - Disease management: the example of amyotrophic lateral sclerosis. AB - Disease management is defined as any medical or pharmaceutical intervention designed to improve both outcomes for the patient and overall cost-effectiveness of the health plan. Disease management focuses on the patient throughout the entire course of the disease, involving both health providers and third-party payers. It requires structured management of change, inter- and intra professional communication and access to information, identification of pertinent economic and clinical outcomes, and the establishment of guidelines, computerized systems and quality assurance. The concept of disease management remains controversial, primarily because its effectiveness is untested. Furthermore, if only economic outcomes are considered, ethical problems such as the selection of populations (for example, the exclusion from health care of people deemed 'too old') will emerge. As a model of neurodegenerative disease, amyotrophic lateral sclerosis (ALS) is a suitable condition for disease management. Many possible targets for disease management initiatives in ALS can be defined, including training, communication, education, guidelines for diagnosis, follow-up, clinical trials and treatments. Medico-economic studies need to be improved if accreditation is planned. Much remains to be done to improve the therapy and disease management of ALS. However, the identification of optimal treatment will improve care of ALS patients, particularly in less affluent countries. PMID- 9747931 TI - Intracellular localization of viral RNA in Eimeria necatrix of the domestic fowl. AB - The intracellular localization of viral RNA in three different stages of Eimeria necatrix, namely, the first- and second-generation meronts and the macrogamonts, were examined at the light microscopy level by in situ hybridization. Digoxigenin labeled riboprobes generated from partial cDNA clones from 5.6-kb and 4.5-kb dsRNA (pzenv1 and pzenv2) were used in this study. Viral RNA was found to be confined to the cytoplasm of the eimerian host; no viral RNA was detected in chicken tissue. The intense hybridization signal observed in the cytoplasm of the immature meronts with the SP6 riboprobe of pzenv1 or the T7 riboprobe of pzenv2 was probably due to their hybridization to positive strands that had been extruded from the virus during transcription. In mature meronts the intensity of the signal is lower, signifying a decrease in transcription activity. Viral replicase activity may thus be synchronized with the growth phase of the eimerian host. PMID- 9747932 TI - Immunoelectron microscopic evidence for release of eosinophil granule matrix protein onto microfilariae of Onchocerca volvulus in the skin after exposure to amocarzine. AB - The involvement of eosinophils in the host reaction to microfilariae (mf) of Onchocerca volvulus was studied by immunohistochemistry and immunoelectron microscopy. Skin biopsies were obtained from patients after transepidermal administration of the microfilaricide amocarzine. At 20-28 h after the application of amocarzine, mf were degenerated or dead and a marked eosinophil parasite adherence (EPA) reaction was seen, with intense staining for intra- and extracellular eosinophil granule proteins such as eosinophil cationic protein (ECP) surrounding the mf. Immunoelectron microscopically the eosinophil granule matrix in intact and necrotic eosinophils was specifically labeled, whereas granules whose matrix had dissolved showed no specific gold particle binding. As specific labeling was seen on lowly electron-dense material adjacent to matrix depleted granules, the material was regarded as released eosinophil granule matrix material. Intact and necrotic eosinophils, matrix-containing as well as matrix-depleted eosinophil granules, and released eosinophil granule matrix material were observed on the surface of damaged mf and between collagen fibers. The coincidence of mf degeneration, EPA reaction, and release of eosinophil granule matrix material on damaged mf and collagen fibers indicated a role of eosinophils and eosinophil granule matrix protein in the host reaction to mf after amocarzine application. PMID- 9747933 TI - Differential expression of the estrogen-regulated proto-oncogenes c-fos, c-jun, and bcl-2 and of the tumor-suppressor p53 gene in the male mouse chronically infected with Taenia crassiceps cysticerci. AB - Chronic infection with Taenia crassiceps cysticerci produces a 200-fold increase in serum estradiol levels in male mice. The aim of this study was to investigate the expression pattern of c-fos and c-jun, two estradiol-regulated genes, as well as that of p53 and bcl2 in the testes, spleen, and thymus of male mice infected with T. crassiceps cysticerci. In parasitized animals the c-fos mRNA content was significantly increased in all tissues studied, whereas the c-jun mRNA content was increased only in the thymus. The p53 mRNA content was markedly reduced in all tissues of the parasitized animals analyzed, whereas bcl-2 gene expression was abolished in the thymus. On the other hand, thymic cell analysis performed by flow cytometry showed a diminution in the content of CD3+, CD4+, and CD8+ subpopulations in the parasitized mice. Our results suggest that the increase in estradiol levels of the host should change the expression pattern of several genes that participate in apoptosis regulation in the thymus of male mice during chronic infection with T. crassiceps cysticerci. PMID- 9747934 TI - The peroxidoxin 2 protein of the human parasite Onchocerca volvulus: recombinant expression, immunolocalization, and demonstration of homologous molecules in other species. AB - The peroxidoxin protein of the filarial parasite Onchocerca volvulus (OvPXN-2) belongs to a group of highly conserved antioxidant molecules. For a more detailed characterization of this protein and for determination of its expression pattern the OvPXN-2 protein was recombinantly expressed as a His-tagged protein. Under reducing conditions the recombinant protein had an apparent molecular mass of 28 kDa. Considering the size of the His-tag and the FLAG epitope introduced to the recombinant protein, this size is in agreement with that of the native protein identified in O. volvulus extract. Antiserum raised against the recombinant protein was used for immunolocalization. In O. volvulus the antigen is predominantly expressed in the hypodermis and particularly the lateral and median chords show high levels of expression. The protein is also expressed strongly in the hypodermis of infective larvae and more weakly in microfilariae. Related cross-reacting proteins were detected in several Onchocerca species and other filariae. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis in combination with Western blotting revealed proteins with almost identical mobility in extracts prepared from O. ochengi, O. gibsoni, and Dirofilaria immitis. PMID- 9747935 TI - Identification of the 150-kDa surface antigen of Entamoeba histolytica as a galactose- and N-acetyl-D-galactosamine-inhibitable lectin. AB - A monoclonal antibody that reacts with a 150-kDa protein of Entamoeba histolytica on Western immunoblotting under nonreducing conditions inhibits the adherence and cytotoxicity of the ameba to mammalian cells in vitro. Affinity purification of solubilized trophozoites using the monoclonal antibody and electrophoresis yielded three glycoproteins with molecular masses of 150, 170, and 260 kDa, suggesting the existence of either a common epitope or the close association of these proteins. The 260-kDa fraction was identified as the well-known galactose (Gal)- and N-acetyl-D-galactosamine (GalNAc)-inhibitable lectin. The 150- and 170 kDa fractions seemed to exist as part of a 380-kDa native protein with an isoelectric point of pH 6.9. The N-terminal amino acid sequence of the 150-kDa protein was unique, indicating that the protein was not a degraded product of the 260-kDa lectin. By gel filtration, the 260-kDa lectin and the 150/170-kDa protein could be separated. When Chinese hamster ovary cells were pretreated with the fraction consisting of the 150/170-kDa protein the adherence of trophozoites to Chinese hamster ovary cells was competitively inhibited to a level equivalent to that observed for the 260-kDa lectin. The inhibitory effect was lost in the presence of Gal and GalNAc but was not influenced by the presence of glucose. These results demonstrate that the 150/170-kDa protein is a Gal/GalNAc inhibitable lectin. The existence of a sugar-binding domain in the protein was confirmed by Gal-affinity chromatography. PMID- 9747936 TI - Toxoplasma gondii: an ultrastructural study of host-cell invasion by the bradyzoite stage. AB - The invasion of Toxoplasma gondii tachyzoites and bradyzoites was followed in bovine kidney cells via electron microscopy. The process of invasion differed between bradyzoites and tachyzoites. In the early stages of entry there was evidence of localised formation of membrane projections in the host cell adjacent to the parasite. Parasite reorientation and rhoptry release appeared to be necessary for invasion; however, the tight junction could not be clearly discerned and there was no evidence of constriction or of any membrane shedding from the parasite. The resulting parasitophorous vacuole was smaller than the tachyzoite vacuole and parasites were frequently found to lie immediately under the host cell membrane. The vacuole was rapidly adapted by the release and formation of an intra-phagosomal membrane network, while the parasitophorous vacuole formed a relationship with host-cell endoplasmic reticulum. PMID- 9747937 TI - Superoxide dismutase and total antioxidant status of larvae and adults of Trichostrongylus colubriformis, Haemonchus contortus and Ostertagia circumcincta. AB - Superoxide dismutase (SOD), a cytosolic enzyme that is specific for scavenging superoxide radicals, is involved in protective mechanism(s) in tissue injury following oxidative processes and phagocytosis. The presence of SOD activity in larval and adult Trichostrongylus colubriformis, Haemonchus contortus and Ostertagia circumcincta was examined using a xanthine-xanthine oxidase assay and by polyacrylamide gel electrophoresis (PAGE) and non-denaturing sodium dodecyl sulfate (SDS)-PAGE followed by specific enzyme staining. Total antioxidant status was determined using the Randox Laboratories kit. The infective larval stages (L3) of the three species contained 8-10 times more activity than the corresponding adults. SOD activity from adult parasites was sensitive to KCN and SDS and may therefore belong to a Cu/Zn and Mn class of enzymes. SOD from the larvae was sensitive only to KCN, suggesting that it may belong to a Cu/Zn class of enzymes. Insignificant interspecies variation was observed when SOD isozyme profiles of larvae were compared. PAGE showed at least five bands of SOD activity with molecular weights of between 18 and 205 kDa. Examination of total antioxidant status showed that non-enzymatic antioxidant potential was also present, but only in the infective larvae. The level of antioxidants in the three genera of larvae studied was similar and amounted to about 0.33-1.07 microM/mg of protein. PMID- 9747938 TI - Development of an enzyme-linked immunosorbent assay for detection of antibodies to Babesia bigemina in cattle. AB - Monoclonal antibodies, directed against a 58-kDa Babesia bigemina merozoite antigen that reacted strongly with immune sera from experimentally and naturally infected cattle in Western blots, were used to develop a competitive-inhibition enzyme-linked immunosorbent assay (ELISA). As based on the testing of 70 antibody positive sera from experimentally infected cattle and 166 antibody-negative sera collected in non-endemic areas of Australia, the sensitivity and specificity of the ELISA were 95.7% and 97.0%, respectively. In sequential sera collected from six calves during the course of experimental B. bigemina infections the ELISA detected seroconversion at about 10 days post-inoculation. The specificity of the ELISA was not affected by the presence of antibodies to B. bovis, Anaplasma marginale or Theileria buffeli. In 42 sera from cattle experimentally infected with B. bovis but negative for B. bigemina the specificity of the ELISA was 95.2%. The use of a competitive-inhibition ELISA format detecting only antibody directed against a single epitope on the 58-kDa antigen appears to have overcome many of the specificity problems that have plagued serological tests for B. bigemina in the past. The test should be useful for epidemiology studies, particularly in areas where B. bovis and B. bigemina have overlapping distributions. PMID- 9747939 TI - Echinococcus multilocularis: relationship between susceptibility/resistance and liver fibrogenesis in experimental mice. AB - To analyze collagen and other matrix protein deposits in experimental alveolar echinococcosis as well as the expression of lysyl oxidase, the enzyme that initiates the first steps in the pyridinoline cross-linking of collagen, and to establish a relationship between resistance/susceptibility to Echinococcus multilocularis larval growth and fibrogenesis, we compared AKR/J mice (susceptible to E. multilocularis infection) with NMRI mice (resistant hosts) in this study. Collagen deposits in the lesions were evaluated using a colorimetric method; the nature of matrix proteins involved in the periparasitic fibrosis and lysyl oxidase expression were assessed using immunostaining on tissue sections. The results obtained in this sequential study confirm that fibrogenesis is an important aspect of the host immune reaction against parasitic development and that both the extent and the course of matrix protein deposition differ in the liver of susceptible and resistant mice, respectively. The long-lasting expression of alpha-actin and lysyl oxidase by host cells in NMRI mice suggests that in this resistant strain, fibrosis was not only more developed but also more highly cross-linked and, thus, less sensitive to collagenases than in susceptible mice. A very strong expression of lysyl oxidase by parasitic cells was observed in both strains of mice; the observation that E. multilocularis itself has a role in lysyl oxidase cross-linking of host collagens can be hypothesized and would be a new example of parasite-host interplay. PMID- 9747940 TI - Murine schistosomiasis mansoni: experimental analysis of bone marrow and peripheral myelopoiesis. AB - In schistosomiasis a systemic hyperplasia of the monomacrophagic cell lineage is associated with its mild modifications in myelograms and hemograms. We monitored the in vitro proliferation of myeloid precursors obtained from bone marrow, blood, spleen, and liver. The macrophage colony-forming unit (M-CFU) numbers were stable in bone marrow but increased progressively in spleen and in liver, reaching in each organ the values equivalent to one femur. The bone marrow had an increased production and enhanced capacity to release M-CFU. Their quantitative increase in blood and in peripheral tissues of schistosome-infected mice was associated with their qualitative modifications: augmented proliferative capacity, enhanced adhesion, and accelerated differentiation. The accelerated release of monomacrophage progenitors and their enhanced proliferation in peripheral tissues potentially account for the relatively low involvement of the bone marrow and for an efficient in situ production of phagocytes, which participate in host reactions to parasites. PMID- 9747941 TI - Stable and unstable environments influence the genetic diversity of the nematode Teladorsagia circumcincta, a parasite of small ruminants. AB - Acclimation of animal parasitic nematodes in the laboratory is the result of environmental disturbance; moderate numbers of infective larvae are introduced into and develop in a few naive hosts (versus many hosts with a resistance status to parasite infection under natural conditions), and stable conditions such as convenient moisture and temperature (versus the unstable climatic environment in the field) are offered to the free-living stages. The acclimation of sheep and goat lines of the nematode Teladorsagia circumcincta in lambs was arranged in the following putative order of increasing disturbance: sheep line and high success of experimental infection, sheep line and poor success of infection, goat line and high success of infection, sheep line with very poor success of infection, goat line and poor success of infection, and sheep isofemale line with founder and inbreeding effects. The genetic variability was assessed using the enzymes glucose-phosphate isomerase, lactate dehydrogenase, malate dehydrogenase, mannose phosphate isomerase, and phosphoglucomutase, in starch-gel electrophoresis. The ranking on increasing Fst values (increasing genetic differentiation) observed between initially introduced and twice-passaged generations ranged from 0.003 (sheep line in sheep with high infective success) to 0.19 (sheep isofemale line) and matched to a certain extent with disturbance. The introduction of a goat line in sheep was a major disturbance, whereas in sheep lines the major factor of variation was due to the founder effect, i.e., the effective number of nematodes introduced to seed the acclimated lines. The deficiency in heterozygotes, which remains largely unexplained, was not modified during acclimation. In most cases the introduction of worms from nature resulted in lower overall genetic variability in the subsequent laboratory-reared populations. PMID- 9747942 TI - Improved in vitro cultivation of larvae of the bovine lungworm Dictyocaulus viviparus. AB - Parasitic larvae of Dictyocaulus viviparus are of major importance for the development of immunity in cattle. The conditions for in vitro cultivation of D. viviparus larvae as well as their morphology have thus far been only poorly investigated. Exsheathed larvae were cultivated in vitro in RPMI-1640 (Gibco-BRL, pH 7.2) containing 20% newborn Calf serum, 200 U Moronal/ml, and 200 U penicillin/streptomycin/ml. Incubation was performed at 39.5 degrees C at 0, 5%, 10%, and 20% CO2. Average development rates to third-moult (3 M) or fourth-stage (L4) larvae at 5% CO2 incubation were 8.33% (SD +/- 7.76%), 22.52% (SD +/- 13.09%) at 10% CO2, and 38.01% (SD +/- 15.63%) at 20% CO2. These differences were statistically significant. Some morphological features of these larvae are described. PMID- 9747943 TI - The role of type-1 and type-2 T-helper immune responses in HIV-1 vaccine protection. AB - The dichotomy of type-1 and type-2 T-helper (Th) immune responses is thought to be an obstacle to develop Human immunodeficiency virus-type- (HIV-1) vaccines capable of inducing effective cellular as well as humoral immune responses. Macaca mulatta were immunized using two different HIV-1sf2 envelope vaccine strategies, based on either immune-stimulating complexes (ISCOM) or chimeric Fowlpox (FP) vaccines. One month following the third immunization all animals were heterologously challenged with simian/human immunodeficiency virus (SHIVsf13). Vaccinated monkeys, which were protected had the highest levels of both type-1 and type-2 HIV-1 specific T-helper cell (Th) responses in addition to the highest homologous and heterogenous virus neutralizing antibodies. To determine how long Th responses persisted and if they correlated with protection, animals were rechallenged after waiting for four months without re-boosting. Macaques which maintained the highest gp120-specific type-1 (IFN-gamma) responses were protected, while there was evidence of viral clearance in two others. These findings demonstrate the importance of both or mixed type-1 and type-2 Th responses in HIV-1 vaccine induced immunity while suggesting a possible role of persistent type-1 responses in maintaining protective immunity over time. PMID- 9747944 TI - Chimeric SHIV that causes CD4+ T cell loss and AIDS in rhesus macaques. AB - By animal to animal passage in rhesus and pig-tailed macaques, we developed a rhesus model of HIV-1 disease in humans. Rhesus macaques infected with a cell free stock of SHIVKU-2 developed CD4+ T cell loss, primary lentiviral encephalitis and pneumonia, and AIDS. Six of nine rhesus macaques died within eight months post-inoculation, while the remaining three are at five, five, and eight months post-inoculation, respectively. Animals infected by either mucosal or parenteral routes of infection had a similar course of infection. PMID- 9747945 TI - Presence of circulating CTL induced by infection with wild-type or attenuated SIV and their correlation with protection from pathogenic SHIV challenge. AB - The aim of this study was to evaluate the role of CTLs in the protection from challenge with pathogenic SHIV in macaques vaccinated with attenuated virus. More specifically, we have analyzed the CTL response in cynomolgus macaques vaccinated/infected with the attenuated SIVmacC8 or the wild-type SIVmacJ5 and correlated these responses to the protection from SHIV89.6P challenge. SIVmacC8 vaccinated monkeys demonstrated a broader CTL response than the SIVmacJ5-infected animals. Nevertheless, CTL against some proteins in SIVmacC8-vaccinated monkeys became progressively more difficult to detect through the day of challenge. In regards to protection from superinfection with SHIV89.6P, neither the presence of circulating CTL nor the CTL precursor frequency against any of the tested proteins correlated with the outcome of the challenge when SIVmacJ5- and SIVmacC8 infected animals were analyzed together. By analyzing the SIVmacC8-vaccinated animals separately, only the protected animal had detectable CTL precursors with moderate frequencies against all three tested proteins at the day of challenge. PMID- 9747946 TI - Sequence diversity of SIV(Mne) Nef in vivo and in vitro. AB - We have compared nef gene sequences isolated by PCR from peripheral blood lymphocyte DNA of macaques which had been inoculated with either biologically or molecularly cloned SIV(Mne). Two samples from each animal obtained either early after infection (week 2-8) or after significant CD4+ depletion (week 21-137) were analyzed. Three substitutions in the predicted Nef amino acid sequence were seen in all animals at the late time point, and two more in all but one. Two of the common exchanges are located about 40 residues apart in the Nef core sequence, but are in proximity on the tertiary structure as judged by computer modelling using the structure of the HIV Nef core protein as a guide. Most recurring in vivo changes replaced a residue found in the cloned Nef sequence with one present in a consensus derived by aligning the Nef sequences of the SIVsm/HIV-2 groups. Animals inoculated with virus already containing the "late version" nef gene developed a more aggressive disease. The macaque adapted (MA)nef conferred a threefold higher infectivity to the cloned virus, but had no effects on CD4 downregulation. Propagation of virus with MAnef in tissue culture resulted in the rapid emergence of variants with newly attenuated nef. These findings suggest that the selective pressure on nef in vivo and in vitro are different. PMID- 9747947 TI - In vitro infection of primate PBMC with simian/human immunodeficiency virus, SHIV(SF33A): correlation to in vivo outcome. AB - The macaque/SIV animal system is an important model for studying AIDS pathogenesis and for evaluating the efficacy of vaccines and anti-viral therapeutics. However, differences between HIV-1 and SIV envelope proteins exist that render the SIV/macaque model of limited value when examining envelope determinants of retroviral pathogenesis. To overcome this problem, we utilized a chimeric virus, SHIV(SF33), containing the env gene from HIV-1SF33 in the context of the molecular clone SIVmac239, in the macaque animal model. In this study SHIV(SF33A), a pathogenic virus that evolved in vivo from a rhesus macaque infected intravenously with the molecular clone SHIV(SF33) was used in both in vitro and in vivo studies. By using a cell culture system, we examined the biological properties of our parental and animal-adapted chimeric viruses and compared in vitro susceptibility to in vivo studies. PMID- 9747948 TI - Natural history of SIVmac BK28 and H824 infection in Macaca nemestrina. AB - The natural histories of disease progression induced by two closely related molecular clones of SIVmac were evaluated to determine the utility of these viruses for modeling fast and slow progression to AIDS in Macaca nemestrina. Viral and immune parameters were measured to determine differential progression. Survival time, viral load and CD4+ T cell decline all were indicative of distinct rates of progression, while early measurements of interferon-gamma (IFNgamma) producing cells did not indicate significant differences. PMID- 9747949 TI - SIVrcm infection of macaques. AB - In a prior report, we described the isolation and characterization of SIVrcm, a distinct primate lentivirus found in a household pet Red-Capped Mangabey (RCM) in Gabon. SIVrcm is divergent from HIV-1 and HIV-2/SIV families of primate lentiviruses. In this report, additional in vitro replication studies and the results of SIVrcm infection in macaques are presented. SIVrcm causes little cytopathic effedct in Molt 4 Clone 8 cells and in rhesus and human PBMCs. In vivo, SIVrcm is non-pathogenic after 200 days in rhesus macaques and after one year in cynomolgous macaques, but does cause a chronic infection in both macaques. PMID- 9747950 TI - In vitro HIV-1 infection in Macaca nemestrina PBMCs is blocked at a step beyond reverse transcription. AB - Various stages in the lifecycle of HIV-1 were investigated in Macaca nemestrina and humans in vitro. Early events were analyzed by end point dilution DNA PCR with HIV-1 and SHIV infected PBMCs, while p24 and p27 ELISA assays were used to analyze core antigen production from infected cells. The results demonstrate that a step in the virus life cycle, beyond reverse transcription is blocked for HIV-1 infection in macaque cells. PMID- 9747951 TI - Longitudinal analysis of behavioral, neurophysiological, viral and immunological effects of SIV infection in rhesus monkeys. AB - A model is proposed in which a neurovirulent, microglial-passaged, simian immunodeficiency virus (SIV) is used to produce central nervous system (CNS) pathology and behavioral deficits in rhesus monkeys reminiscent of those seen in humans infected with human immunodeficiency virus (HIV). The time course of disease progression was characterized by using functional measures of cognition and motor skill, as well as neurophysiologic monitoring. Concomitant assessment of immunological and virological parameters illustrated correspondence between impaired behavioral performance and viral pathogenesis. Convergent results were obtained from neuropathological findings indicative of significant CNS disease. In ongoing studies, this SIV model is being used to explore the behavioral sequelae of immunodeficiency virus infection, the viral and host factors leading to neurologic dysfunction, and to begin testing potential therapeutic agents. PMID- 9747952 TI - Expression and in vitro evaluation of rhesus macaque wild type (wt) and modified CC chemokines. AB - Several human CC chemokines have been shown to inhibit HIV/ SIV infection in vitro, providing the rationale for their potential use in vivo. However, because of their inherent physiological effect, such chemokines are reasoned to be of limited therapeutic value due to potential side effects. The knowledge that amino terminus modified or deleted human RANTES retains its receptor binding properties but loses its signaling properties has provided a means to use such modified chemokines in vivo for possible therapeutic benefits. In efforts to test the efficacy of such modified chemokines, our laboratory has cloned, sequenced, and prepared recombinant forms of wild-type (wt) and amino-terminus modified rhesus macaque chemokines MIP-1alpha, MIP-1beta, and RANTES. These sets of chemokines were tested for their potential to inhibit SIV infection and induce signaling. The data showed that whereas wt chemokines retained both virus inhibitory and signaling functions, corresponding amino-terminus modified chemokines only showed virus inhibitory effects without detectable signaling effects. Such reagents will be valuable for evaluation of their therapeutic potential in vivo, either alone or as adjuncts to other chemotherapeutic drugs. PMID- 9747953 TI - Dendritic cells from skin and blood of macaques both promote SIV replication with T cells from different anatomical sites. AB - The SIV-macaque system offers the opportunity to study the pathogenesis and immune aspects of a primate retroviral infection in which immunodeficiency also develops, much like HIV infection in humans. Since it is known that human dendritic cells (DCs) are involved in HIV replication, mature cytokine-generated DCs obtained from precursors in the blood and skin-derived DCs were isolated from healthy rhesus macaques and compared with respect to their ability to support SIV infection. Here, it is shown for both skin- and blood-derived DCs that i) virus production depends on both DCs and T cells, ii) this occurs similarly with T cells from blood, skin, spleen, or lymph nodes, and iii) DCs can transmit virus equally to syngeneic and allogeneic T cells. No differences between DCs from skin or blood were observed. Therefore, the easily accessible blood-derived DCs of macaques provide an appropriate population to study the role of DCs in immunodeficiency virus infection. PMID- 9747954 TI - Intracellular cytokine expression in the CD4+ and CD8+ T cells from intestinal mucosa of simian immunodeficiency virus infected macaques. AB - Isolated intraepithelial lymphocytes (IEL) and lamina propria lymphocytes (LPL) from jejunum of SIV infected animals were examined for alterations in basal cytokine expression by RT-PCR. Remarkable changes in IFNgamma and IL-10 RNA levels were observed in IEL and LPL in SIV infection while IL-4 and IL-2 RNA levels remained unaltered. In addition, the CD4+ and CD8+ LPL were examined for intracellular cytokine production following mitogenic activation by flowcytometry. Both CD4+ and CD8+ T lymphocytes in intestinal mucosa retained the potential to produce IFNgamma in response to mitogenic stimulation in vitro, without a remarkable change in IL-4 production. The dominant IFNgamma cytokine response could be one of the major contributing factors in SIV associated enteropathy. PMID- 9747955 TI - Down-modulation of the ZAP-70 protein tyrosine kinase in macaque T lymphocytes infected with SIVsmmPBj14. AB - The simian immunodeficiency virus SIV-PBj14 is the most virulent primate lentivirus identified to date. Other SIV strains, including the parental SIVsmm9, require mitogen-activated peripheral blood mononuclear cells (PBMC) for replication in vitro; however, SIV-PBj14 replicates in quiescent pig-tailed macaque PBMC and induces cellular proliferation, consistent with its in vivo pathogenesis. To identify mechanisms involved in SIV-PBj14-induced T-cell proliferation, kinases important in early T-cell receptor-mediated activation pathways were studied. Immunoblot analyses showed that ZAP-70 protein, a tyrosine kinase, was downregulated, primarily in CD8+ T cells, as early as 30 minutes after in vitro infection of quiescent macaque PBMC with SIV-PBj 14. Furthermore, this downregulation required the presence of either CD4+ T cells or adherent cells or both cell populations. In agreement with the in vitro results, ZAP-70 expression was downregulated in macaque PBMC, spleen, and rectal lymph node cells as early as 2 days after rectal inoculation of pig-tailed macaques with SIV PBj14. This phenomenon, however, was not observed in cells obtained from distal lymph nodes to which the virus had not disseminated, implying that the presence of SIV-PBj14 is necessary to induce downregulation of ZAP-70. PMID- 9747956 TI - Myocardial metabolism studied during warm blood antero-retrograde reperfusion in ischaemic human hearts. AB - We propose modified warm blood antegrade-retrograde reperfusion (WBARR) of arrested hearts as a metabolic model with which to study substrate exchange and energy metabolism during the recovery phase after 90 min of ischaemia in man. Eleven anaesthetized patients undergoing aorto-coronary bypass were studied during WBARR. The protocol was designed as follows: period 1, a warm blood reperfusion with potassium (3 min); period 2, a warm blood reperfusion without potassium (2 min). The perfusion flow rate averaged 250+/-2 ml/min at the beginning of period 1 and 218+/-19 ml/min at the beginning and at the end of period 2; the perfusion was performed antegradely and retrogradely in the arrested hearts. Samples were simultaneously taken from the coronary venous sinus (CVS) and from the aortic root needle (AR). At the beginning of WBARR lactate release was 85+/-44 micromol/min and at the end it had significantly decreased to 21+/-99 micromol/min (P<0.03). Simultaneously, non-esterified fatty acids (NEFA) and beta-hydroxy-butyrate were initially released (71+/-61 and 22+/-66 micromol/min, respectively), while at the end of the WBARR there was an uptake of both NEFA (20+/-22 micromol/min; P<0.01) and beta-hydroxy-butyrate (12+/-35 micromol/min; P=0.290). Alanine, glycerol and branched chain amino acid balance across the heart did not significantly change. In summary after 90 min of ischaemia the heart energy metabolism is mainly anaerobic and based on glucose consumption, with lactate, NEFA and amino acids, which are mainly released. After 5 min of WBARR (recovery from ischaemia), lactate release is significantly reduced and NEFA becomes the energy supply of the heart. In conclusion, (1) WBARR is a valuable method with which to study myocardial metabolism in anaesthetized humans and may be combined with the use of tracers; (2) the study of myocardial metabolism in arrested hearts eliminates the imprecisions arising from the noncontinuous coronary blood flow; (3) NEFA become an important source of energy utilized by human hearts in the recovery phase from ischaemia. PMID- 9747957 TI - Environmental modulation of D-fructose insulinotropic action. AB - At concentrations in excess of about 80 mmol/l, D-fructose stimulates insulin release from rat islets incubated in the absence of any other exogenous nutrient, an optimal secretory response being recorded in the 240 to 320 mmol/l range. D galactose and 3-O-methyl-D-glucose fail to reproduce the insulinotropic action of D-fructose. At a concentration of D-fructose close to the threshold value for such an insulinotropic action (80 mmol/l), as little as 1.0-4.0 mmol/l D-glucose is sufficient to increase insulin release, with a sigmoidal concentration response relationship similar to that otherwise evoked by much higher concentrations of the aldohexose. The release of insulin evoked by D-fructose (240 mmol/l) is abolished in the absence of Ca2+ or presence of KCN (2.0 mmol/l), partially inhibited by 3-O-methyl-D-glucose (80 mmol/l) or D-mannoheptulose (1.0 mmol/l), and potentiated by forskolin (10 micromol/l), theophylline (1.4 mmol/l), cytochalasin B (21 micromol/l) and glibenclamide (5 micromol/l). These findings indicate that the stimulation of insulin release by high concentrations of D fructose corresponds to an active secretory process modulated by the metabolic fate of the hexose, the availability of ATP, the activity of ATP-sensitive K+ channels, the extracellular concentration of Ca2+, the cell content in cyclic AMP and the motile events under the control of the microfilamentous cell web. PMID- 9747958 TI - Elevated hepatocyte growth factor in sera from patients with insulin-dependent diabetes mellitus. AB - Hepatocyte growth factor (HGF) is a pleiotropic cytokine known to be involved in tissue regeneration and repair. We measured serum levels of HGF in patients with insulin-dependent diabetes mellitus (type 1). The patients were divided into four groups: (1) 10 patients at clinical presentation before insulin treatment; (2) 19 patients with newly diagnosed type 1 diabetes (diabetes duration 1/2-3 years); (3) 14 patients with long-standing type 1 diabetes without renal involvement (diabetes duration >10 years, and urinary albumin excretion (UAER) <20 microg/ min); and (4) 20 patients with long-standing type I diabetes with renal involvement (diabetes duration >10 years and UAER 20-500 microg/min). Sera from 24 age- and sex-matched healthy blood donors constituted a control group. The HGF levels of the four groups were (mean +/- SD); group 1, 0.74+/-0.14; group 2, 0.78+/-0.40; group 3, 0.86+/-0.42; group 4, 0.79+/-0.27 ng/ml, compared to 0.43+/ 0.24 ng/ml in the control group (P<0.0008). HGF levels were not significantly different between the four patient groups. The elevated serum HGF levels did not correlate with complications related to type 1 diabetes, such as UAER, retinopathy and macrovascular complications, suggesting that HGF levels were not associated with the type 1 diabetes complications. In conclusion, our results show that type 1 diabetic patients have increased serum HGF levels compared with controls and that HGF is elevated to the same extent in newly diagnosed as well as in long-standing type 1 diabetes. PMID- 9747959 TI - Hypoglycaemic symptoms described by diabetic children and their parents. AB - Hypoglycaemic events are frequent complications of insulin-dependent diabetes mellitus in children. The signs and symptoms referred to by young children can be difficult to understand and often seem to be different from those described by their parents. We analysed the hypoglycaemic symptoms described by a group of patients and their parents. We studied 40 pairs consisting of a parent and a diabetic child by using a structured questionnaire with 27 items concerning different symptoms of hypoglycaemia. The mean+/-SD age of the children was 10.4+/ 2.4 years, with duration of disease 6.2+/-2.1 years and their HbA1c was 8.2 2.0%. For the statistical analysis we used the principal component analysis. All the children followed a multiple injection regimen. The frequency and intensity of the hypoglycaemic signs described by patients and parents were similar both for neuroglycopenic (uncoordination, confusion, odd behaviour, dizziness) and autonomic symptoms (trembling, sweating, pounding heart, hunger). Moreover, our questionnaire showed a high frequency of behavioural changes. In conclusion, from the analysis of the questionnaires collected, we found that both parents and children gave almost the same score to the symptoms observed. This means that there is a concordance between the symptoms reported by the children and those reported by their parents. PMID- 9747960 TI - Relationship between non-enzymatic glycosylation and changes in serum insulin like growth factor-1 (IGF-1) and IGF-binding protein-3 levels in patients with type 2 diabetes mellitus. AB - The possible occurrence of increased non-enzymatic glycosylation of serum insulin like growth factor binding protein-3 (IGFBP-3) in vivo and the changes that would simultaneously occur in serum levels of IGFBP-3 and insulin-like growth factor-1 (IGF-I) were investigated. We measured levels of IGF-I and IGFBP-3 and the degree of glycation of total serum protein and IGFBP-3, in serum samples obtained from patients with poorly controlled non-insulin-dependent diabetes (type 2) and from age-matched non-diabetic controls. Type 2 diabetic patients had significantly higher glycated serum protein (GlyP) levels. GlyP significantly correlated with age in the control (r = 0.315, P<0.05) but not in the type 2 diabetes group. Control and diabetic subjects had comparable serum IGF-I levels and in both groups IGF-I levels tended to decrease with age (r = -0.567, P<0.001 and r = 0.465, P<0.05 for control and type 2 diabetic subjects, respectively). In the type 2 diabetes group, IGF-I levels showed a negative correlation with serum GlyP values (r = -0.476, P<0.05). Type 2 diabetic and control patients had comparable serum IGFBP-3 levels, which were significantly higher in diabetic patients in the older age subgroups. A negative correlation was found between IGFBP-3 levels and age in the control (r = -0.705, P<0.001) and in the type 2 diabetes groups (r = 0.463, P<0.05). A significant negative correlation was found between IGFBP-3 levels and GlyP in control (r = -0.449, P<0.002) but not in type 2 diabetic subjects. The mean glycated IGFBP-3 (GlyIGFBP-3) levels were higher in the oldest type 2 diabetic patients. In these patients, GlyIGFBP-3 was negatively associated with IGF-I levels (r = -0.447, P<0.05). The IGF-I/IGFBP-3 molar ratio was significantly reduced in the 46-60-year-old type 2 diabetic group, whereas the IGF-I/IGFBP-3 ratio was positively and significantly correlated with GlyP levels only in the control group (r = 0.489, P<0.01). Our results show that: a) increased non-enzymatic glycosylation of IGFBP-3 occurs in vivo; and b) this effect is accompanied by an increase in IGFBP-3 levels. These results suggest that the IGF-I/IGFBP-3 system is another target for the metabolic derangements of type 2 diabetes. Its alterations might play a role in diabetic complications. PMID- 9747961 TI - Residual beta-cell function and spontaneous clinical remission in type 1 diabetes mellitus: the role of puberty. AB - To investigate the role of puberty on spontaneous clinical remission and on secretion of residual C-peptide during the first year of type 1 diabetes mellitus, we studied 77 pre-pubertal, 39 pubertal and 41 post-pubertal type 1 diabetic patients. Spontaneous partial clinical remission (HbA1c within the normal range and insulin dose less than 0.3 U x kg(-1) body weight x day(-1) lasting for at least 10 days) decreased with duration of diabetes: months 3 vs 6 vs 12, respectively 13 vs 7 vs 4% (P<0.025). Remission was higher in post pubertal than pubertal and prepubertal patients: month 6 respectively 20 vs 5 vs 1% (P<0.001). Secretion of C-peptide was significantly lower in pre-pubertal than the other two groups of patients. Basal and stimulated C-peptide secretion were higher in patients in clinical remission than in those who were not: basal value 0.4 (0.26-0.53) vs 0.28 (0.14-0.4) nmol/l (P<0.05); stimulated value 0.63 (0.5 0.95) vs 0.56 (0.31-0.74) nmol/l (P<0.05). Spontaneous remission is less frequent in children and adolescent patients than in adult post-pubertal patients, but different mechanisms may be involved. Low residual insulin secretion seems implicated in children meanwhile low insulin sensitivity could be more important in pubertal patients. PMID- 9747962 TI - Effect of doxazosin in mild to moderate hypertensive patients with insulin dependent diabetes mellitus. AB - Diabetic patients often develop hypertension, and the presence of both hypertension and diabetes doubles the risk of death from coronary heart disease (CHD). Moreover, the presence and importance of abnormalities such as high low density lipoprotein (LDL) cholesterol and triglycerides levels as CHD risk factors in insulin-dependent diabetes mellitus type 1 have been downplayed, while increasing evidence suggests that the management of type 1 patients should include control of dyslipidemia and hyperglycemia and an effective antihypertensive treatment able also to reduce risk factors for coronary artery events. In this study we assessed the antihypertensive and metabolic effects of doxazosin in hypertensive patients with type 1 diabetes. We show that the drug normalizes blood pressure, and while no improvement in glucose control was observed, it reduced total cholesterol and increased HDL cholesterol as well as the HDL to total cholesterol ratio. The changes of the various parameters studied, including the calculated CHD risk score based on the Framingham equation, suggest that doxazosine can reduce the CHD risk for hypertensive type 1 patients. PMID- 9747963 TI - Effects of glucagon-like peptide-1 (7-36)amide on insulin stimulated rat skeletal muscle glucose transport. AB - Glucagon-like peptide-1 binding sites have been reported in peripheral tissues including muscle. However, the potential extra-pancreatic effects of glucagon like peptide-1(7-36)amide are controversial. To evaluate whether glucagon-like peptide-1(7-36)amide has any effects on skeletal muscle glucose transport, isolated rat soleus muscles were incubated in increasing concentrations of insulin (0-150 nmol/l) in the presence or absence of 1 nmol/l glucagon-like peptide-1(7-36)amide for 3 h. Subsequently glucose transport was measured as uptake of [3H]-O-methylglucose. It was found that glucagon-like peptide-(7 36)amide has a small but significant stimulating effect on skeletal muscle glucose transport independent of the insulin concentration (P<0.01). However, because of the magnitude of the observed effect, the physiological importance of glucagon-like peptide-1(7-36)amide on skeletal muscle glucose metabolism is questionable. PMID- 9747964 TI - Reproducibility of erythrocyte sodium-lithium countertransport activity and ambulatory blood pressure measurements in type 1 diabetes mellitus. AB - We aimed to study the reproducibility of sodium-lithium countertransport [SLCT] activity and ambulatory blood pressure monitoring [ABPM] in type 1 diabetes. We did this by performing repeated measurements of SLCT activity and ABPM in 11 recent-onset diabetic children and in 11 patients with longer duration of diabetes. Both parameters were related to microalbuminuria. In the older group of diabetic children a significant correlation [r = 0.78; P<0.005] in SLCT activity between the first and second study was observed [514.3+/-186.4 vs 491.0+/-148.0 micromol/l erythrocytes/h]. Diurnal systolic and diastolic blood pressure were comparable at both time points within the same group of diabetic children [in group 1: 102.6+/-6.1 vs 108.6+/-7.6 mmHg N.S.; in group 2: 113.4+/-10.6 vs 114.0+/-7.8 mmHg N.S. Diastolic blood pressure in group 1: 57.4+/-4.8 vs 65.7+/ 6.9 mmHg N.S., in group 2: 70.6+/-9.1 vs 68.5+/-5.3 mmHg N.S.]. Moreover, there was a significant correlation in both diurnal and nocturnal systolic blood pressure between the first and second study in the whole diabetic population. Both SLCT activity and blood pressure values obtained by ABPM were found to be reproducible individual characteristic markers in type 1 diabetic children. PMID- 9747965 TI - Crosses between diabetic BB/OK and wild rats confirm that a third gene is essential for diabetes development. AB - Several crossing studies with diabetic BB rats have shown that in addition to the lymphopenia (Iddm1) and the MHC class II genes of the RT1U haplotype (Iddm2) there are further non-MHC genes essential for diabetes development. Because diabetes-resistant inbred rat strains may be homozygous for one of the diabetogenic non-MHC genes, masking the expression of diabetogenic genes and leading to an underestimation of the number of diabetogenic genes, we crossed wild and diabetic BB/OK rats. The F1 hybrids were backcrossed onto diabetic female (BC1W-F, n=97) and male BB/OK rats (BC1W-M, n=98) transferred to a specified-pathogen-free environment and studied for the frequency and age at onset of diabetes up to an age of 30 weeks. Comparing the results of these BC1 W hybrids with similarly derived hybrids using diabetes-resistant DA rats (BC1DA-F, n=113; BC1DA-M, n=216), the diabetes frequency in total was comparable indicating the action of three recessive genes. The percentage of diabetics in Iddm1 and Iddm2 homozygotes confirmed the existence of the third gene, Iddm3, but there were some sex differences; significantly more male than female BC1W-F and significantly more BC1DA-M than BC1DA-F males were diabetic. Regarding the age at onset, the BC1W-F hybrids manifested not only significantly earlier, but also more uniformly than BC1DA-F and BC1-M hybrids. PMID- 9747966 TI - Incidence of type 1 diabetes mellitus in children in Slovenia during the years 1988-1995. AB - The incidence of type 1 diabetes mellitus in Slovenian children aged 0-14 years was studied between 1 January 1988 and 31 December 1995. The crude annual incidence rate of the disease (per 100000) over this 8-year period was 8.00 (95% C. I. 6.98-9.02) for both sexes (7.18 for boys and 8.87 for girls). Thus, the incidence standardized to the world population was 7.59 (95% C. I. 6.57 - 8.61). Male/female ratios were 1.33 in the age group 0-4 years, 0.66 in the age group 5 9 years, and 0.83 in the age group 10-14 years. The study has proven that the incidence of type 1 diabetes in Slovenia is similar to that in other central European countries where the population is of different ethnic origin. However, a remarkably higher incidence of the disease in girls than boys except in the age group below 5 years of age was found which needs further investigation. PMID- 9747967 TI - The molecular karyotype of the megabase chromosomes of Trypanosoma brucei and the assignment of chromosome markers. AB - We present the molecular karyotype of the megabase chromosomes of Trypanosoma brucei stock TREU927/4 (927). We have identified 11 diploid chromosomes ranging in size from 1 to 5.2 Mb approximately and pairs of homologues differ in size by up to 15%. A total of 401 cDNA probes were hybridised to T. brucei stock 927 chromosomes and 168 chromosome-specific markers were defined. Most of these markers were hybridised to the separated chromosomal DNA of two other cloned field isolates and four F1 progeny clones from a laboratory cross. The chromosomes vary in size by up to two and a half times between stocks and the DNA content of the 11 pairs of homologues varies by up to 33% in different stocks. Stock 927 contains the smallest chromosomes and the least nuclear genomic DNA. Nevertheless, all 11 syntenic groups of cDNA probes are maintained in all stocks. In the F1 hybrids only we have identified one extra PFG band to which none of our probes hybridise. We have shown that probes thought to be specific for the bloodstream-form variant surface glycoprotein expression sites hybridise to different chromosomes in different stocks and may hybridise to either one or both of a homologous pair of chromosomes. We have also determined the chromosomal location of the ribosomal RNA gene arrays. PMID- 9747968 TI - Protein prenyl transferase activities of Plasmodium falciparum. AB - Prenylated proteins have been shown to function in important cellular regulatory processes including signal transduction. The enzymes involved in protein prenylation, farnesyl transferase and geranylgeranyl transferase, have been recent targets for development of cancer chemotherapeutics. We have initiated a systematic study of protein prenyl transferases of the malaria parasite, Plasmodium falciparum, to determine whether these enzymes can be developed as targets for antimalarial chemotherapy. We report here the identification of protein farnesyl transferase and protein geranylgeranyl transferase-I in the malaria parasite, P. falciparum. The farnesyl transferase has been partially purified from the cytosolic fraction through ammonium sulfate precipitation and Mono-Q chromatography. Farnesyl and geranylgeranyl transferase-I activities are present at all stages of P. falciparum intraerythrocytic development with maximum specific activity in the ring stage. Geranylgeranyl transferase-I specific activity is two times that of farnesyl transferase in the ring stage. Peptidomimetics and prenyl analogues of protein farnesyl transferase substrates were tested as in vitro inhibitors of partially purified P. falciparum prenyl transferase and of malaria parasite growth. The peptidomimetics were significantly more potent inhibitors than lipid substrate analogues of both the activity of Mono-Q purified enzyme and parasite growth in intraerythrocytic cultures. Exposure of the parasite to the peptidomimetic L-745,631 also showed significant inhibition of morphological development beyond the trophozoite stage. These studies suggest the potential of designing or identifying differential inhibitors of P. falciparum and mammalian prenyl transferases as an approach to novel malaria therapy. PMID- 9747969 TI - A Plasmodium chabaudi protein contains a repetitive region with a predicted spectrin-like structure. AB - cDNA and genomic DNA clones covering the entire open reading frame (ORF) for a Plasmodium chabaudi 96V protein were isolated. From the first ATG codon the intronless gene codes for a 229-kDa protein. Antisera raised against recombinant polypeptides coded by two different regions of the gene reacted with a 240/225 kDa doublet on Western blots of parasite extracts. In immunofluorescence studies the same sera detected the antigen at the apical end of the merozoite, possibly in rhoptry organelles. In Western blotting experiments the recombinant polypeptides were recognised by antibodies induced by natural infection. A 364 amino acid residue repetitive region, based on 32 11-mer repeats divided by two 6 mer repeats into three blocks, is located in the centre of the protein. Analysis of this repetitive region led us to propose a model in which each of the three units forms an alpha-helical coiled-coil triple-helix containing a possible leucine-histidine zipper. Each unit resembles in structure the units present in spectrin. The repeat region is flanked by predicted heptad based alpha-helical coiled-coil regions, and we propose that the protein forms a dimer. The 229-kDa protein has the overall character of a cytoskeletal protein. We have named the 229-kDa protein repetitive organellar protein (ROPE) and suggest that ROPE may be involved in the process of invasion, possibly by interacting with the erythrocyte cytoskeleton, and that the leucine histidine-zipper may be involved in molecular mimicry of spectrin. PMID- 9747970 TI - Expression and cellular localization of Trypanosoma cruzi type II DNA topoisomerase. AB - Topoisomerases are enzymes that participate in many cellular functions involving topological manipulation of DNA strands. There are two types of topoisomerases in the cell: (a) type I topoisomerases; and (b) type II topoisomerases (topo II). Previously we have cloned and sequenced the gene encoding Trypanosoma cruzi topo II (TcTOP2). This study group has raised an antiserum against recombinant type II DNA topoisomerase (TctopoII) to study the expression of this gene during T. cruzi differentiation and to determine the cellular location of the enzyme. Western blot analysis showed that T. cruzi TctopoII is expressed in the replicative epimastigotes but not in the infective and non-replicative trypomastigotes. However, slot blot analysis of RNAs extracted from epimastigotes and metacyclic trypomastigotes showed that the mRNA encoding the enzyme is present in both developmental stages of the parasite. Confocal laser microscopy using the antiserum raised against recombinant TctopoII showed that the enzyme is located exclusively in the nucleus of the parasite. Similar results were obtained by immunofluorescence analysis of Crithidia fasciculata. However, monoclonal antisera against the corresponding enzyme extracted from C. fasciculata recognizes a kinetoplast protein in both T. cruzi and Crithidia. PMID- 9747971 TI - Antifolate resistance due to new and known Plasmodium falciparum dihydrofolate reductase mutations expressed in yeast. AB - Two new dihydrofolate reductase (DHFR) mutations were recently discovered in Plasmodium falciparum samples from an area of Bolivia with high rates of in vivo resistance to pyrimethamine-sulfadoxine: a Cys-->Arg point mutation in codon 50 and a five amino acid insertion after codon 30, termed the Bolivia repeat. We used a yeast expression system to screen these new DHFR mutants, as well as all of the other known DHFR mutant genotypes, against four antifolates: pyrimethamine, cycloguanil, chlorcycloguanil, and WR99210. The prodrug proguanil was also evaluated. The primary 108-Asn mutation, the known secondary mutations 51-Ile, 59-Arg and 164-Leu, as well as the 50-Arg mutation, all progressively enhanced pyrimethamine resistance in naturally observed combinations with one another, with the presence of 164-Leu most significantly increasing resistance. Cycloguanil and chlorcycloguanil resistance were most impacted by 164-Leu and the paired 16-Val/108-Thr. Proguanil had no effect on malaria DHFR. All DHFRs analyzed were sensitive to WR99210. The Bolivia repeat did not markedly affect drug sensitivity. We conclude that malaria DHFR can be reliably, rapidly and inexpensively analyzed in yeast for activity against a broad spectrum of antifolates. This system may be useful for initially characterizing newly discovered genotypes before proceeding to P. falciparum transfection; for large scale geographic surveys of drug resistance; and for screening new antifolates or new antifolate combinations for their effectiveness against a large panel of DHFR mutants. PMID- 9747972 TI - Upregulation of Jun and Fos family members and permanent JNK activity lead to constitutive AP-1 activation in Theileria-transformed leukocytes. AB - Theileria parasitises bovine leukocytes and transforms them into proliferating, metastatic tumours, where the infection resembles a leukaemia-like disease. We have studied the signal transduction pathways leading to activation of the transcription factor AP-1 in different transformed leukocytes. Parasite infection leads to an up-regulation of all members of the Jun/Fos family of proteins and surprisingly, this occurs in the absence of any detectable ERK, or p38 MAP kinase activity. In the parasitised B-sarcoma TBL3, AP-1 induction occurs in the absence of any JNK activity. In contrast, in infected macrophage and B-cell lines, AP-1 transcriptional activity is strictly associated with the parasite-induced constitutive activation of JNK and subsequent c-Jun N-terminal phosphorylation. Thus, constant AP-1 transcriptional activity involves both an upregulation in the levels of Jun and Fos proteins and constitutive JNK activation. PMID- 9747973 TI - The molecular characterization of the major polar tube protein gene from Encephalitozoon hellem, a microsporidian parasite of humans. AB - The microsporidia are obligate intracellular protozoan parasites of increasing importance as human pathogens, which are characterized by a small resistant spore with a single polar filament that coils around the sporoplasm. When stimulated, the polar filament rapidly everts out of the spore to form a hollow polar tube through which the sporoplasm passes, thus serving as a unique mechanism of transmission. A genomic library of the human microsporidium Encephalitozoon hellem was screened using a polyclonal rabbit antibody (anti-PTP Eh55) produced to the major HPLC purified polar tube protein (PTP) of E. hellem. This antibody localized to intrasporal polar filaments and extrasporal polar tubes of E. hellem by immunogold electron microscopy confirming the polar tube specificity of the antibody. A total of 14 anti-PTP Eh55 reactive genomic clones were identified and purified. A PTP gene was identified consisting of 1362 bp coding for 453 amino acids. The N-terminus of the translated protein consists of aputative N-terminal signal sequence of 22 amino acids, which when cleaved results in a mature protein of 431 amino acids with a predicted molecular mass of 43 kDa. The protein has a high proline content (14.6%) and contains a central domain of six alternating tandem repeats of 20 amino acids. After ligation of the gene into a glutathione S transferase (GST) expression vector, a fusion protein was produced that reacted by immunoblotting with the polar tube specific anti-PTP Eh55. The gene was present as a single copy in the genome and there was no homology with other known genes. As the polar tube is a critical structure for the transmission of this organism to a new host cell, further study of PTPs may lead to the development of new therapeutic strategies and diagnostic tests. PMID- 9747974 TI - A cell cycle model for the tachyzoite of Toxoplasma gondii using the Herpes simplex virus thymidine kinase. AB - Toxoplasma gondii (RH strain) tachyzoites were transfected with a plasmid containing a fusion of the chloramphenicol acetyl transferase and the Herpes simplex virus-2 thymidine kinase coding regions and transgenic parasites obtained by chloramphenicol selection. CTK11, a single high expressing clone was isolated based on immunofluorescence and contained approximately five integrated copies of the fusion sequence. Lysates prepared from this clone displayed thymidine kinase activity of 2.9 pmol min(-1) microg(-1) protein, whereas thymidine kinase activity was not detected in lysates from the parental RH strain. Growth of CTK11 tachyzoites was fully inhibited in 5 microM ganciclovir and thymidine and in 2.5 microM 5-bromo-2'-deoxyuridine. While the inhibitory effects of ganciclovir were lethal, low concentrations of thymidine (10 microM) were largely reversible. Asynchronously growing CTK11 tachyzoites were found to contain major G1 (1 N) and S phase (1 N+) distributions as determined by relative propidium iodide fluorescence and with reference to the haploid (1 N) DNA content of a T. gondii sporozoite population. CTK11 tachyzoites blocked 4 h in 10 microM thymidine exhibited mean fluorescence consistent with a 1 N complement of DNA indicating growth was arrested in G1. Following the removal of excess thymidine, parasites immediately entered S phase, thus confirming the late G1 block. Parasites with a 2 N complement of DNA (G2 + M) first appear at 2 h post-release, while 1 N (G1) parasites re-appear at 3 h suggesting the length of S phase is < or = 2 h and that of G2 + M is < or = 1 h. Within 7 h, parasites had transited G2 + M and much of G1 and re-entered S of the subsequent cell cycle--a time consistent with the doubling of these parasites in culture. Thus, the CTK11 tachyzoite cell cycle is similar to those of higher eukaryotic cells and is characterized by major G1 and S phases and a relatively short G2 + M. PMID- 9747975 TI - Conserved organization of genes in trypanosomatids. AB - Trypanosomatids are unicellular protozoan parasites which constitute some of the most primitive eukaryotes. Leishmania spp, Trypanosoma cruzi and members of the Trypanosoma brucei group, which cause human diseases, are the most studied representatives of this large family. Here we report a comparative analysis of a large genomic region containing glucose transporter genes in three Salivarian trypanosomes (T. brucei, T. congolense and T. vivax), T. cruzi and Leishmania donovani. In T. brucei, the 8 kb (upstream) and 14 kb (downstream) regions flanking the glucose transporter genes cluster contain two and six new genes, respectively, six of them encoding proteins homologous to known eukaryotic proteins (phosphatidylinositol 3 kinase, ribosomal protein S12, DNAJ and three small G-proteins--Rab1, YPT6 and ARL3). This gene organization is identical in T. brucei, T. congolense and T. vivax suggesting that Salivarian trypanosomes have a high level of conservation in gene organization. In T. cruzi and Leishmania, the overall organization of this cluster is conserved, with insertion of additional genes when compared with T. brucei. Phylogenetic reconstitution based on glucose transporters is in accord with the monophyly of the genus Trypanosoma and the early separation of T. vivax within Salivarian trypanosomes. On the basis of gene organization, biochemical characteristics of isoforms and phylogeny, we discuss the genesis of the glucose transporter multigene family in Salivarian trypanosomes. PMID- 9747976 TI - Single nucleotide resolution of promoter activity and protein binding for the Leishmania tarentolae spliced leader RNA gene. AB - In Kinetoplastid protozoa, trans-splicing is a central step in the maturation of nuclear mRNAs. In Leishmania, a common 39 nt spliced-leader (SL) is transferred via trans-splicing from the precursor 96 nt SL RNA to the 5' terminus of all known protein-encoding RNAs. In this study, promoter elements of the L. tarentolae SL RNA gene have been identified with respect to transcriptional activity and putative transcription factor binding. We have mapped the essential regions in the SL RNA gene promoter at single nucleotide resolution using both in vivo transcription and in vitro protein/DNA binding approaches. Two regions located upstream of the SL RNA gene were identified: a GN3CCC element at -39 to 33 and a GACN5G element at -66 to -58 were essential for SL RNA gene transcription in stably transfected cells. Consistent with other known bipartite promoter elements, the spacing between the GN3CCC and GACN5G elements was found to be critical for proper promoter function and correct transcription start point selection, as was the distance between the two elements and the wild-type transcription start point. The GACN5G element interacts specifically and in a double-stranded form with a protein(s) in Leishmania nuclear extracts. The degree of this protein DNA interaction in vitro correlated with SL RNA gene transcription efficiency in vivo, consistent with a role of the protein as a transcription factor. The core nucleotides GACN5G fit the consensus PSE promoter structure of pol II-transcribed snRNA genes in metazoa. PMID- 9747977 TI - A small spliceosomal-type intron occurs in a ribosomal protein gene of the microsporidian Encephalitozoon cuniculi. PMID- 9747978 TI - The circumsporozoite protein of Plasmodium chabaudi contains a large, pre-region I repeat domain. PMID- 9747979 TI - Telomeric sequences of Cryptosporidium parvum. PMID- 9747980 TI - Uptake of an antiplasmodial protease inhibitor into Plasmodium falciparum infected human erythrocytes via a parasite-induced pathway. PMID- 9747981 TI - A Plasmodium falciparum hemolytic activity. PMID- 9747982 TI - An upstream regulatory element containing two nine basepair repeats regulates expression of the Entamoeba histolytica hgl5 lectin gene. PMID- 9747983 TI - Development of new photopolymerizable dental sealants. AB - This paper describes the results obtained in the optimization of the composition of dental sealants in relation to the nature and proportions of monomer mixtures and photoinitiating system employed. The quantification and variation of certain parameters which determine the quality of a dental sealant (such as viscosity and penetrating power, residual double bonds, solubility and absorption, volume shrinkage and certain specific mechanical properties) have resulted in the development of new formulations. The composition which has achieved the best results of all the above properties was that corresponding to the monomer mixture bis-GMA/tri(ethylene glycol) dimethacrylate (TEGDMA) 40/60 wt%, and the photoinitiating system camphorquinone (CQ) with co-initiators N,N,3,5 tetramethyaniline (TMA) or N,N-dimethyl-p-toluidine (DMPT) in the ratio 1:1. The final properties and characteristics of the obtained formulations are superior to those of commercial dental sealants currently in use. PMID- 9747984 TI - Effect of surface properties on the antithrombogenicity of silk fibroin/S carboxymethyl kerateine blend films. AB - Polymeric blends of silk fibroin (SF) and S-carboxymethyl kerateine (SCMK) were prepared by the solvent casting method to study the effect of surface properties on the antithrombogenicity. The films of SF/SCMK showed better antithrombogenic properties than SF or SCMK alone. Among them, the film containing 50 wt% SCMK showed the best antithrombogenicity. When the SF/SCMK films were treated with methanol, the antithrombogenicity of the films was scarcely affected except the SF-rich ones. The enhanced antithrombogenic properties were explained in terms of polarity of the surface. The blend films showed an enhancement of polar contribution to surface free energy (gamma(P)S and polar stabilization energy (I(SW)). SF-rich films showed high gamma(P)S and I(SW) values when treated with methanol. This change of surface properties was considered to be due to the fact that the conformational transition from random coil structure to beta-structure of proteins may have affected the surface properties, especially the polar properties. PMID- 9747985 TI - Adhesion of different bacterial strains to low-temperature plasma treated biomedical PVC catheter surfaces. AB - In this study, firstly five different bacteria (i.e. Coagulase positive and negative staphylococcus, Streptococcus pyogenes, Escherichia coli, Pseudomonas aeruginosa) with their different strains were isolated and used. The contact angle, surface free energy, p-xylene adhesion, and zeta potential of these bacteria were in the range of 43-69 deg, 45.4-61.8 erg cm(-2), 2.3-80.3%, and from -650.2 to + 17.5 mV, respectively. Most of the bacteria were negatively charged. Attachment of these bacteria to PVC catheter and its DMAEMA- and AAc plasma treated forms were investigated. Bacterial attachment to the hydrophobic PVC catheter was high. Both plasma treatments caused significant drops in bacterial attachment in most of the cases. The effects of AAc-plasma treatment was more significant. PMID- 9747986 TI - A swine model for the evaluation of efficacy of anti-microbial catheter coatings. AB - A swine model was developed to investigate the efficacy of percutaneous venous catheters with anti-microbial coatings. The catheters used in the study consisted of silver-coated and uncoated catheters, both designed for percutaneous venous access. Five commercial pigs were each implanted with three venous catheters and followed for a period of 90 days. Two of the three catheters were coated and one was uncoated. To evaluate the percutaneous aspects of the catheters in the model, two venous access catheters were implanted percutaneously, parallel to the dorsal midline. These catheters were just caudal to the region that is dorsal to the scapula in each animal. In each case, the catheter to the left of the dorsal midline was silver-coated while the catheter to the right of the dorsal midline was uncoated. A silver-coated catheter was also implanted in the left external jugular vein of each animal and buried subcutaneously in order to evaluate the elution of the coating through the body under venous contact. Over the 90 day period, the concentration of silver in the blood rose to a mean peak level of 23.2 ppb following implantation of the catheters and then decreased after the second post-surgery week. The histological evaluation and macroscopic inspection at necropsy revealed minimal tissue response to both coated and uncoated materials. Data on bacterial growth indicated that bacteria were present at the terminal subcutaneous end of two of the uncoated percutaneous catheters. Based upon serum silver levels, exudate formation, histological examination, and bacterial growth information, the swine model was deemed to be suitable for testing the efficacy of catheters containing anti-microbial coatings. PMID- 9747987 TI - Studies on blood compatibility of terpolymers composed of methyl methacrylate, methoxypolyethyleneglycol methacrylate, and dimethylsiloxane methacrylate. AB - Terpolymers composed of methyl methacrylate (MMA), polydimethylsiloxane methacrylate (PDMSMA), and methoxypolyethyleneglycol methacrylate (MPEGMA), having different compositions were synthesized. Platelets were not adsorbed onto terpolymer surfaces composed of 50 wt% MMA, 25 wt% PDMSMA and 25 wt% MPEGMA, while on terpolymers with the other compositions, platelet adsorption and fibrin clot were observed. It was shown that PDMS segment was predominant on these terpolymer surfaces via XPS. Receding contact angles of terpolymers, on which no platelet was observed, showed intermediate values between PDMS- and MPEG-rich surfaces. It was suggested that these terpolymers had blood compatibility. PMID- 9747988 TI - Modelling the kinetics of antigen-antibody reactions at particle enhanced optical immunoassays. AB - Functionalized latexes coated by antibodies are used in diagnostic tests for the detection of antigens in biological fluids. A simple kinetic model is presented which is related to the optical monitoring of the formation of specific complexes between antigen and antibody amplified by latex beads. The antibodies are chemically coupled onto chloromethylstyrene (CMST) particles. The kinetic model is able to describe the immunoprecipitin curves of immunolatex beads. The number of fitting parameters is relatively reduced (only three), and the meaning of these parameters can be interpreted in terms of the chemical equilibrium constant, the percentage of active IgG on the latex beads, and optical response. The model explains very well the optical response of immunolatex prepared by covalent coupling of antibodies on polymer carriers. PMID- 9747989 TI - Anticoagulant and antiprotease activities of a heparinoid sulfated glucoside bearing polymer. AB - We studied anticoagulant and antiprotease activities of the poly(glucosyloxyethyl methacrylate) sulfate [poly(GEMA)-sulfate] in plasma and purified enzyme systems in order to evaluate the anticoagulant behavior of a heparin-like sulfated glucoside-bearing polymer. As a result, we found that poly(GEMA)-sulfate can inhibit some coagulation proteases, although its antiprotease behavior differed from those of heparin and dextran sulfate. Poly(GEMA)-sulfate could not enhance antithrombin activity; therefore, we did not observe any significant inhibition of Factor Xa via antithrombin. However, we found that poly(GEMA)-sulfate was able to inhibit thrombin through the activation of heparin cofactor II. In addition, poly(GEMA)-sulfate was able to inhibit Tenase. In our previous research. we found that the anticoagulant activity of poly(GEMA)-sulfate is due primarily to the formation of an insoluble complex with fibrinogen. This paper showed that the antiprotease activities of poly(GEMA)-sulfate contribute to some extent to its anticoagulant activity. PMID- 9747991 TI - Comparison of bone regeneration in a rabbit skull defect by recombinant human BMP 2 incorporated in biodegradable hydrogel and in solution. AB - The objective of this study is to compare bone regeneration induced by recombinant human bone morphogenetic protein-2 (rhBMP-2) incorporated into a biodegradable gelatin hydrogel with that by rhBMP-2 in aqueous solution. After treating rabbit skull defects of 6 mm diameter with the two rhBMP-2 dosage forms, both of them increased the bone mineral density (BMD) at the skull defects with implantation time to a significantly higher extent than a rhBMP-2-free aqueous solution and a rhBMP-2-free empty gelatin hydrogel (p < 0.05). There was no quantifiable difference in BMD between the two dosage forms of rhBMP-2. Histological examination revealed that the integrity of newly generated bone increased with the rhBMP-2 dose, irrespective of the dosage form. The bone defect was filled with regenerated bone 21 days after treatment. PMID- 9747992 TI - The Kund Jansen lecture: Amputee rehabilitation--finding the niche. PMID- 9747990 TI - Effect of polyelectrolyte complex (PEC) on human periodontal ligament fibroblast (HPLF) functions in the presence of glucocorticoids. AB - Cell functions in vivo are stimulated by extracellular matrices, vitamins, growth factors, and hormones. In this paper, the effects of glucocorticoids, dexamethasone (Dex), and Cortexrone (Cor) on the growth and differentiation of human periodontal ligament fibroblast (HPLF) were discussed in relation to a polyelectrolyte complex (PEC) consisting of polysaccharides (chitin, cellulose derivatives, and chitosan) as a tissue-culture material. A Dex-treatment at a concentration of 10(-)-10(-7) M inhibited one-half of HPLF growth in comparison with 10(-9) M Dex-treatment and no additive medium and produced aggregates on the chitosan-sulfated chitin PEC (SPECs) with regard to the degree of sulfate substitution. On the chitosan-sulfated cellulose PEC, 10(-7)-10(-9) M Dex treatment promoted HPLF growth and inhibited the production of aggregates. On the other hand, a Cor-treatment, a mineral corticoid, which inhibits the interaction between Dex and its receptor, increased HPLF growth on SPEC141, but the HPLF did not construct aggregates. A Dex and Cor mixture-treatment inhibited one-third HPLF growth in comparison with 10(-5) M Dex-treatment and produced aggregates on PEC. The cooperative effect of both the culture material and hormones was found to control HPLF growth and morphology. The alkaline phosphatase (ALPase) activities of HPLF increased with an increase in the Dex and Cor concentration. The value of Dex-treated HPLF ALPase activity demonstrated a two-fold increase from that with Cor-treatment. The ALPase activity of Dex and Cor mixture-treated HPLF on PEC decreased with an increase in the Cor concentration, because Cor increased HPLF growth on PEC. In using carboxymethylated chitin derivatives as the polyanion, HPLF decreased in cell growth and produced aggregates in the absence of the additives, suggesting that PEC induces HPLF differentiation using only the stimulation of the material surface. PMID- 9747993 TI - The influence of prior stroke on the prosthetic rehabilitation of lower limb amputees. AB - Concurrent stroke is believed to have an adverse influence on the process and outcome of prosthetic rehabilitation, but there is limited published evidence for this. The aim of this study was to establish a clearer picture in order to assist decision making for both patients and professionals. Demographic and clinical data were collected from all lower limb amputees referred from North and West Yorkshire for prosthetic rehabilitation. Additional data were collected from all new lower limb amputees in three of the referring health districts, irrespective of prosthetic referral. Patients with prior stroke were less likely to be referred for prosthetic rehabilitation. Improved mobility and independence were seen following prosthetic rehabilitation irrespective of prior stroke. The group with prior stroke compared well with the non-stroke group in terms of walking aid usage, but a smaller proportion of the stroke group were able to walk 30 m without stopping and there were trends for smaller gains in independence in the stroke group. Nevertheless, this study demonstrates that prosthetic rehabilitation can be successful in a selected amputee population with prior stroke. In those who continue prosthetic use for one year, outcome is similar to that in patients without stroke. PMID- 9747994 TI - Comparison of the lightweight Camp Normal Activity Foot with other prosthetic feet in trans-tibial amputees: a pilot study. AB - Clinically relevant information regarding the useability of prosthetic feet is scarce. The industry is not obliged to perform clinical studies before marketing the product. Clinicians however are limited in their possibilities (organisation and finance) to determine the useability of a technical product. This small study is an example of how in general the useability of a technical product is established in clinical practice. The Camp Normal Activity Foot (CNAF), a carbon prosthetic foot, was compared objectively and subjectively with a number of other prosthetic feet (same price bracket) in three subjects with trans-tibial (TT) amputations. The CNAF is low in weight and has favourable stiffness and hysteresis properties. The stiffness of the pylon of the CNAF seems to be limited as also is the possibility of adaptation of the CNAF. The CNAF distinguishes itself, in this study, but not convincingly with respect to the energy consumption in walking, the step-time parameters (symmetry) and the subjective judgement of the users. An additional virtue of the CNAF seems to be its light weight. PMID- 9747995 TI - Morphological changes during early trans-tibial prosthetic fitting. AB - Morphological changes in the amputation stump may have serious implications regarding the suspension and fit of the prosthetic socket. In an earlier study (Lilja and Oberg, 1997) the authors have shown that the volume of the trans tibial amputation stump decreases according to a negative power function after amputation, and that the stump volume does not stabilise until four months after the operation. In the present study, Magnetic Resonance Imaging (MRI) technique was used to examine morphological changes in the amputation stump after trans tibial amputation in a small number of cases. The authors expected to find a decrease in the cross-sectional area of the stump and of the separate muscles similar to the findings in earlier studies. However, two different patterns were found. The cross-sectional area of the entire stump as well as that of the medial muscle group changed according to the authors' hypothesis, i.e. an initial fast decrease, followed by a more moderate decrease of the area. In the lateral muscle group another pattern was found. After an initial rapid decrease the area increased, sometimes to a magnitude larger than the initial value. After the amputation the lateral muscle group may acquire a new function, contributing to the suspension of the socket. Despite the limited number of patients, this study presents findings which may be important in the clinical fitting of trans-tibial prostheses. PMID- 9747996 TI - An "appropriate technology" trans-femoral prosthesis, using materials available in Nepal. AB - The use of western components and materials for prostheses is prohibitively expensive in most developing countries. In addition, local customs and conditions vary considerably from those for which the prostheses were designed. For these reasons, a trans-femoral prosthesis has been developed in Pokhara, Nepal, using entirely locally available materials, and with a view to fulfilling local requirements as far as possible. This paper describes the materials and fabrication technique for the component parts of the prosthesis, the local conditions for which it was developed, and a three year follow-up of the first prosthesis issued. Only one serious design fault was discovered during this period, and a modification to the fabrication procedure introduced. The authors believe that if this trend continues, this style of prosthesis may be useful in the future for Nepali amputees and perhaps also in other countries, particularly where mass production of components is not practical. PMID- 9747997 TI - Conventional versus microchip controlled pneumatic swing phase control for trans femoral amputees: user's verdict. AB - A questionnaire survey of 22 selected transfemoral amputees who were switched from pneumatic swing phase control knee joints (PSPC) to microprocessor controlled intelligent knee joints (IP) is presented. On overall rating all respondents considered the IP to be an improvement or a great improvement compared to the PSPC and none decided to revert back or wished to use their previous PSPC prosthesis on a regular basis. However the IP was not rated to be superior in every area of the questionnaire. Walking at different speeds, walking further, reduction of energy consumption were the main areas where subjective improvements were perceived by the amputees themselves. It is strongly believed that patients' own views should be an important and integral part of the evaluation of new prosthetic technology. PMID- 9747998 TI - An audit of the quality of the stump and its relation to rehabilitation in lower limb amputees. AB - An audit was undertaken amongst the lower limb (adult) amputees, 60 unilateral transfemoral (TF) and 72 unilateral trans-tibial (TT), who attended a Disablement Services Centre (DSC) during a one year period, to determine whether amputees with better quality stumps (as assessed by a scoring system used at the Centre) achieve better outcome from prosthetic rehabilitation and whether there is any relation between the construction of the stumps and the grade of surgeons. At eighteen months (minimum follow up of six months) there were 31 (52%) TF and 54 (75%) TT amputees wearing prostheses. Some 44 amputees with Grade A stumps (score of 60 and over, out of a possible 100) needed 154 days to achieve the predicted mobility grade, 15 (34%) of them needed alteration of prosthesis, attended the Centre every 42 days and achieved the activity score of -25.7; 41 amputees with Grade B stumps (scores less than 60) needed 206 days to achieve the predicted mobility grade, 24 (58.5%) of them needed alteration of prosthesis, attended the Centre every 29 days and achieved the activity score of -39.1 (less active than Grade A). The trainee surgeons (registrars, staff grade surgeons and SHOs) produced 26 Grade A stumps out of 67 amputations (40%) and the Consultants and the Senior Registrars (senior team) produced 37 Grade A stumps out of 65 amputations (57%). However, only 36% of amputees were prescribed prostheses at their first attendance (60% Grade A, and 40% Grade B). PMID- 9747999 TI - Criteria for prosthetic provision: "he who pays the piper calls the tune". AB - Previously instituted policies regarding prosthetic limb provision had been deemed dependable. A follow-up home visit study showed that 18 of 60 patients that had been provided with prostheses, did not make use of them. Analysis showed that three categories of patients made up the large majority of the non-users; double amputees, blind persons and those with psychiatric disorders. In order to attempt to eliminate the wastage of prosthetic provision to non-ambulators a new policy decision was made. Doubtful ambulators and those from the three aforementioned categories will be initially provided with temporary prostheses. Only after a period of months of temporary prosthetic usage at home will a decision be made as to whether a permanent prosthesis will be issued. PMID- 9748000 TI - Powered prosthetic hands in very young children. AB - Myoelectric prostheses are generally not provided in the United Kingdom for children before the age of 3 1/2 years. Following the introduction of a smaller sized electric hand in the United Kingdom in 1993 the authors decided to introduce electrically powered hands for a group of congenital upper limb deficient children at a much younger age compared to normal practice. Eleven children were introduced to powered prosthetic hands at an average age of 20.6 months. At the review carried out for the purpose of this paper. 72.7% of these children appeared to be successfully using these powered prostheses. Fitting these young children with powered prostheses and encouraging acceptance and operation of the prostheses appeared to be much less of a problem than might have been anticipated. The parents of all these children have very much liked the introduction of powered hands at this early age and have contributed positively to the prosthetic programme. The authors' experience suggests that introduction of a powered prosthesis at a much earlier age can be a more suitable alternative than provision of a body-powered prosthetic device while waiting to reach an older age before a powered prosthesis is considered. PMID- 9748001 TI - Low-bandwidth telemedicine for remote orthotic assessment. AB - A model for performing remote orthotic assessments using low-bandwidth computer communication technology (video conferencing) was developed, tested, and evaluated. System evaluation involved comparing a series of remote assessments with on-site assessments. While most on-site and on-line results were similar, discrepancies which occurred were attributed to between-clinician differences, measurement technique differences, technical and learning obstacles at the start of the project, and within subject variations during the day. On-line assessment efficiency improved with each on-line session and corresponded with increased confidence in the system, easier system use, and better overall satisfaction. An on-line debriefing session was held with all project clinicians. These clinicians supported continued use of the communication system for rehabilitation consultation and education. Clinically. preliminary face-to-face meetings and a regular practice schedule were recommended. Technically, the system was considered good; however, suggested improvements included using a high quality speaker-phone system, streamlining the video capture process, and providing more reliable telecommunication connections. PMID- 9748002 TI - The essentiality of histo- and cytochemical studies of skeletal muscle in the investigation of neuromuscular disease. 1962. PMID- 9748003 TI - Faiths that failed? Professionalism, technology, and science in 20th-century America. PMID- 9748004 TI - Recombinant human nerve growth factor and diabetic polyneuropathy. PMID- 9748005 TI - Hereditary neuropathy with liability to pressure palsies: a patient's point mutation in a mouse model. PMID- 9748006 TI - Touching and timing consciousness. PMID- 9748007 TI - Amino acids and the brain: too much, too little, or just inappropriate use of a good thing? PMID- 9748008 TI - Practice parameter: Stroke prevention in patients with nonvalvular atrial fibrillation. Report of the Quality Standards Subcommittee of the American Academy of Neurology. PMID- 9748009 TI - Prevention of stroke in patients with nonvalvular atrial fibrillation. AB - OBJECTIVE: To review the risk and pathogenesis of stroke associated with nonvalvular atrial fibrillation (AF) and the efficacies and risks of stroke prevention strategies. BACKGROUND: About 16% of ischemic strokes are associated with AF; AF is an independent risk factor for stroke. METHODS: Review of the literature, focusing on 13 randomized trials of antithrombotic therapy. RESULTS: The overall risk of stroke in AF patients averages about 5%/y, but with wide variation depending on the presence of coexistent thromboembolic risk factors. AF patients with low (about 1% per year), moderate (about 3% per year), and high (about 6% per year) stroke risks have been identified, but the generalizability of risk stratification schemes to clinical practice has not been fully assessed. AF patients with prior stroke or transient ischemic attack, even if remote, are at highest risk (about 12% per year). Adjusted-dose warfarin (target International Normalized Ratio [INR] 2-3) is highly efficacious for preventing stroke in AF patients (about 70% risk reduction) and is safe for selected patients, if carefully monitored. Aspirin has a modest effect on reducing stroke (about 20% risk reduction). The numbers of AF patients that would need to be treated with warfarin instead of aspirin for 1 year to prevent one ischemic stroke are about 200, 70, and 20 for those with low, moderate and high risk, respectively. CONCLUSIONS: Many patients with nonvalvular AF have substantial rates of ischemic stroke. Stratification of stroke risk identifies AF patients who benefit most and least from lifelong anticoagulation. Warfarin is recommended for high-risk AF patients who can safely receive it. Aspirin may be indicated for those with a low stroke risk and for those who cannot receive warfarin. For AF patients considered to have a moderate risk of stroke, individual bleeding risk during anticoagulation and patient preference should particularly influence the choice of antithrombotic prophylaxis. PMID- 9748010 TI - Interferon beta in the treatment of multiple sclerosis: mechanisms of action. AB - Interferon beta (IFN-beta) has been shown in several clinical trials to have efficacy in MS. Its mechanism of action, however, remains unclear. In this review, several biological activities of IFN-beta are highlighted, including its inhibitory effects on proliferation of leukocytes and antigen presentation. Furthermore, IFN-beta may modulate the profile of cytokine production toward that of the anti-inflammatory phenotype, and this appears to occur in the systemic circulation and within the CNS. Finally, IFN-beta can reduce T-cell migration by inhibiting the activity of T-cell matrix metalloproteinases. These activities are likely to act in concert to account for the mechanism of IFN-beta in MS. PMID- 9748011 TI - Cerebral amyloid angiopathy: prospects for clinical diagnosis and treatment. AB - This article reviews diagnosis of cerebral amyloid angiopathy (CAA) during life and possible approaches to prevention. A clinical diagnosis of "probable CAA" can be made in patients aged 60 years or older with multiple hemorrhages confined to lobar brain regions and no other cause of hemorrhage. Gradient-echo MRI facilitates diagnosis by showing previous hemorrhages with high sensitivity. This technique can also mark the progression of CAA, as 50% of studied patients developed new petechial hemorrhages during 1.5 years of follow-up. The apolipoprotein E epsilon2 and epsilon4 alleles are associated with increased risk and earlier age of first hemorrhage, but are neither sensitive nor specific for CAA. The major remaining challenges are to develop new markers for the presence of CAA and treatments to block vascular amyloid deposition and vessel breakdown. PMID- 9748012 TI - Recombinant human nerve growth factor in the treatment of diabetic polyneuropathy. NGF Study Group. AB - BACKGROUND: Preclinical studies have demonstrated that nerve growth factor may prevent or reverse peripheral neuropathy. We have therefore tested the effects of recombinant human nerve growth factor in patients with diabetic polyneuropathy. METHODS: A total of 250 patients with symptomatic diabetic polyneuropathy randomly received either placebo or one of two doses of recombinant human nerve growth factor for 6 months. Patients were assessed for symptoms and signs of polyneuropathy before and after treatment. RESULTS: Compared with placebo, recombinant human nerve growth factor led to significant improvement after 6 months of treatment, as measured by the sensory component of the neurologic examination, two quantitative sensory tests, and the impression of most subjects that their neuropathy had improved. Three prospectively identified multiple endpoint analyses indicated improvements in the nerve growth factor treatment groups over the placebo group in all three analyses (p = 0.032; p = 0.008; p = 0.005). Recombinant human nerve growth factor was well tolerated, with injection site discomfort reported as the most frequent adverse event. CONCLUSIONS: Recombinant human nerve growth factor appears to be safe and shows preliminary evidence of efficacy in patients with symptomatic diabetic polyneuropathy. PMID- 9748014 TI - Genotypic-phenotypic variations in a series of 65 patients with familial amyloid polyneuropathy. AB - OBJECTIVE: To investigate the genotypic-phenotypic variations in a series of patients with familial amyloid polyneuropathy (FAP). BACKGROUND: Progress in molecular genetics has led to the identification of point mutations in the transthyretin (TTR) gene in FAP--a dominantly inherited neuropathy with a fatal outcome. These findings have modified the management of patients with small-fiber neuropathy and allow genetic counseling. METHODS: We performed a clinical and molecular genetic study with screening of the TTR gene mutations and associated haplotypes in 65 patients from 29 unrelated families of French ancestry. RESULTS: We detected nine heterozygous point mutations segregating with FAP. Fourteen families (48%) carried the common methionine (Met) 30 substitution. Seven kindreds (24%) had previously unreported TTR variants, namely asparagine 35, serine (Ser) 91, phenylalanine (Phe) 77, and Ser 116. At least two different haplotypes were associated with each of the following: Met 30, Phe 77, and valine 107, suggesting that multiple founders occurred for each variant. Only 35% of the index patients had affected relatives. Other patients had a sporadic presentation. All progressed to a severe sensorimotor and autonomic neuropathy with frequent cardiac involvement (80%). On average, a late age at onset (54.3 +/ 13.3 years) and a disease duration shorter than 10 years were observed for virtually all variants. CONCLUSION: The heterogeneity of the TTR variants, the late age at onset, and the short duration of the disease found in our patients contrast with the presentation of FAP in Portugal. These findings must be taken into account in the management of both patients and asymptomatic carriers. PMID- 9748013 TI - A novel PMP22 point mutation causing HNPP phenotype: studies on nerve xenografts. AB - BACKGROUND: Hereditary neuropathy with liability to pressure palsies (HNPP) in most cases is caused by a deletion in chromosome 17p11.2-12 or, rarely, mutations resulting in a functional loss of one copy of the peripheral myelin protein 22 (PMP22) gene. Point mutations that lie deep within transmembrane (TM) domains causing major structural changes in PMP22 are associated with severe neuropathy. METHODS: A 25-year-old asymptomatic woman with a normal neurologic examination volunteered as a control subject. Electrophysiologic studies showed multiple entrapment neuropathies, prompting a search for a genetic defect. In addition, sural nerve fascicles from the subject were grafted into the cut ends of the sciatic nerve of nude mice and studied at 2, 6, and 8 weeks and compared with controls. RESULTS: Direct sequencing of the PMP22 gene revealed a G-->A transition at position 202 in axon 3 of the PMP22 gene. To determine if this was a causative mutation rather than a polymorphism, 102 DNA samples from controls were studied; none showed a similar base pair change. In the nerve xenografts, there was a marked delay at the onset of myelination and an impairment in the regenerative capacity of the nude mice axons engulfed by the mutant human Schwann cells. The axon tips were enlarged and demonstrated neurofilament density increase. Neurofilament density distribution histograms were bimodal in xenografts as well as in the subject's sural nerve. CONCLUSION: This study provides unequivocal evidence that a base pair change causing a Val30Met substitution at the junction of the first TM domain and the extracellular loop of PMP22 results in the HNPP phenotype. PMID- 9748015 TI - Denervation of eccrine glands in patients with familial amyloidotic polyneuropathy type I. AB - OBJECTIVE: To study the alterations in the structure and innervation of eccrine glands in familial amyloidotic polyneuropathy (FAP) type I with Val 30 Met transthyretin mutation. BACKGROUND: Anhidrosis of the distal lower limbs is a prominent feature of FAP type I. METHODS: Qualitative and morphometric study of amyloid deposition, eccrine glands, and their innervation in nine patients with FAP type I (duration of sensory symptoms, 8.4 +/- 3.9 years [mean +/- SD]; range, 3 to 15 years) and seven control subjects. RESULTS: On light microscopy, the endoneurium of cutaneous nerve fascicles had no definite amyloid deposition. Amyloid deposition was observed around eccrine glands in seven of nine patients. On electron microscopy, no focal destruction and degeneration of eccrine glands or ducts and of Schwann cell processes with or without nerve terminals or unmyelinated axons were observed in relation to adjacent amyloid deposition. Secretory vacuoles and granules of dark cells were markedly decreased in some secretory coils. Nerve terminals and unmyelinated axons of eccrine glands were considerably fewer in patients than in control subjects, and denervation was prominent in all patients. A few nerve terminals and unmyelinated axons of eccrine glands were present in patients who had experienced sensory symptoms for 3, 5, and 6 years, but were absent in patients with sensory symptoms for more than 7 years. CONCLUSIONS: Eccrine glands are markedly to totally denervated in patients with FAP type I and chronic sensory symptoms. The extent of denervation indicates the severity of autonomic denervation and therefore may suggest the timing of liver transplantation. PMID- 9748016 TI - Physiology of somatosensory perception: cerebral lateralization and extinction. AB - OBJECTIVE: To demonstrate the effects of cerebral lateralization and temporal dynamics on somatosensory perception. BACKGROUND: We postulated that perceptual thresholds for simple somatosensory stimuli would be less in the left than the right hand, and that a left/right asymmetry in extinction would exist in healthy right-handed subjects (but not in left-handed subjects). During the course of these experiments we also examined the controversy concerning the temporal dynamics of somatosensory perception. METHODS: A total of 126 healthy subjects (age range, 6 to 73 years) participated in the study. Effects of handedness, age, vigilance, gaze, and temporal interval on somatosensory perception were examined in a series of experiments. Brief electric pulses were applied to the index finger of one or both hands. RESULTS: Perceptual thresholds are lower in the left than the right hand of healthy right-handed subjects in a large cohort across a wide age range. Left-handed subjects have no overall asymmetry. Even after compensation for baseline threshold differences, single stimuli in right-handed subjects are perceived more readily in the left than the right hand, and left hand targets are more difficult to mask. Leftward eye/head gaze lowers thresholds in both hands of right-handed subjects (compared with right or straight gaze). Extinction was consistently maximal when the mask followed the target by 50 to 100 msec. CONCLUSIONS: The findings demonstrate clearly that left/right perceptual thresholds for simple somatosensory stimuli are asymmetric in healthy right-handed subjects. Both central and peripheral asymmetries exist. The central asymmetry and gaze effects are consistent with right cerebral dominance for externally directed attention. Access of somatosensory stimuli to conscious awareness is delayed and particularly vulnerable to disruption at 50 to 100 msec after onset of the stimulus. PMID- 9748017 TI - The incidence of dementia: a meta-analysis. AB - OBJECTIVE: To carry out a meta-analysis of the age-specific incidence of all dementias, including AD and vascular dementia. BACKGROUND: Several meta-analyses have been carried out on dementia prevalence, but none on its incidence. METHODS: We used loess-curve fitting to analyze data from 23 published studies reporting age-specific incidence data. RESULTS: The incidence of both dementia and AD rose exponentially up to the age of 90 years, with no sign of leveling off. The incidence rates for vascular dementia varied greatly from study to study, but the trend was also for an exponential rise with age. There was no sex difference in dementia incidence (p = 0.21), but women tended to have a higher incidence of AD in very old age, and men tended to have a higher incidence of vascular dementia at younger ages. East Asian countries had a lower incidence of dementia than Europe (p = 0.0004), and also tended to have a lower incidence of AD. CONCLUSIONS: The incidence of dementia rises exponentially to the age of 90 years. Any sex differences are small, and incidence is lower in East Asia than in Europe. PMID- 9748018 TI - Polymorphism of the prion protein is associated with cognitive impairment in the elderly: the EVA study. AB - BACKGROUND: Little is known about the role of the prion protein (PrP(sen)/gene PRNP). PRNP knockout mice studies suggest that PrP(sen) may be involved in CNS degeneration. This observation prompted us to examine the influence of PRNP genetic variability on cognitive abilities in the elderly. METHODS: In a community-based sample of 1,163 subjects aged 59 to 71 years, we characterized the valine (Val) and methionine (Met) allele of the PRNP polymorphism at codon 129. The effect of this polymorphism was estimated on the Mini-Mental State Examination (MMSE) and on a global composite score built from a battery of nine different neuropsychological tests. The results were adjusted for age, gender, education, and apolipoprotein E (apoE) polymorphism. RESULTS: Cognitive impairment (MMSE score < 24) was present in 2.5% of the Met-Met individuals, 2.9% of the Met-Val individuals, and 7.0% of Val-Val subjects (p = 0.02). Subjects homozygous for the PRNP Val allele had a lower MMSE and global score than the two other genotypes (p < 0.003). This effect was of the same magnitude as that of the apoE epsilon4 allele on cognitive performances. Both apoE epsilon4 and PRNP Val allelic effects were additive. CONCLUSION: This observation suggests that variability of the PRNP locus may be associated with cognitive performance in the elderly. This result, if confirmed, offers potential clues for the role of PRNP in the human brain. PMID- 9748019 TI - Serum interferon beta-1a (Avonex) levels following intramuscular injection in relapsing-remitting MS patients. AB - BACKGROUND: The pharmacokinetics of IFNbeta-1a in MS patients are poorly understood. We have previously reported an ELISA sensitive and specific for measuring serum IFNbeta-1b levels in patients with MS. OBJECTIVE: We describe an ELISA to measure interferon beta-1a (Avonex) in the serum of MS patients following IM administration. METHODS: We have developed an ELISA for detecting serum IFNbeta-1a in MS patients receiving 6 million units (MU) of IFNbeta-1a, IM once weekly. The specificity of this ELISA was confirmed by the lack of cross reactivity with other cytokines except for IFNbeta-1b. RESULTS: Serum IFNbeta-1a levels were measured at 3 and 6 months after initiating treatment with IFNbeta-1a in 10 MS patients. At 3 months, all 10 patients had detectable levels ranging from 68 to 86 IU/mL. At 6 months, IFNbeta-1a could be detected in the serum of all but three patients, with levels ranging from 64 to 81 IU/mL. A kinetic study of IFNbeta-1a serum levels in a separate group of six MS patients who had been receiving IFNbeta-1a for several months was carried out. Blood was drawn before and 2, 4, 6, 8, and 24 hours after IFNbeta-1a injection. Peak serum IFNbeta-1a levels were observed at 8 hours and became undetectable at 24 hours after injection. CONCLUSION: The described ELISA may have useful clinical applications in examining the correlation between serum IFNbeta-1a levels and clinical efficacy. PMID- 9748020 TI - Major histocompatibility complex class II alleles and the course and outcome of MS: a population-based study. AB - BACKGROUND: The major histocompatibility complex (MHC) has been consistently associated with susceptibility to MS and the course of several other human autoimmune diseases. A putative association between the course and severity of MS and the MHC remains controversial. METHODS: DR and DQ genotyping by either restriction fragment length polymorphism or sequence-specific PCR-based typing in 119 patients representing 73.4% of the population with MS evaluated in a cross sectional disability survey and 100 healthy controls from Olmsted County, Minnesota. RESULTS: We found a positive association between MS susceptibility and the DR15-DQ6 and DR13-DQ7 haplotypes, and we found a negative association with the DR1-DQ5 haplotype. We found a trend to a positive association of primary progressive MS with DR4-DQ8 and DR1-DQ5 and an association of "bout onset" MS with DR17-DQ2. We did not find an association with disease severity, as defined by EDSS/duration. CONCLUSION: Lack of consistency between different studies may be due to regional variation in MS and limitations of power but likely indicate a minor effect of MHC class II genes on the course and severity of MS. PMID- 9748021 TI - A systematic study of oligodendrocyte growth factors as candidates for genetic susceptibility to MS. French Multiple Sclerosis Genetics Group. AB - OBJECTIVE: To test 23 genes coding for growth factors and their receptors as candidates for MS genetic susceptibility in 84 multiplex families of French origin by linkage analysis. BACKGROUND: Epidemiologic studies have indicated that genetic susceptibility in MS exists. To identify MS susceptibility genes, association and linkage studies were performed with candidate genes suggested by the pathology of MS. The most consistent result was genetic association and linkage of MS to human leukocyte antigen (HLA) DR15. Recent advances in the knowledge of MS pathology have suggested that the oligodendrocyte, the myelin forming cell in the CNS, and its growth factors might play a crucial role in MS. METHODS: Fifty-two polymorphic markers within or flanking 23 candidate genes were used. Data were analyzed with the maximum likelihood score (MLS) approach. We also searched for a genetic interaction with HLA. RESULTS: Negative results were obtained for all candidate genes. The lower limits of the relative risk (Xs) possibly excluded for any candidate gene ranged from 1.3 to 2.8. Positive MLS values (up to 0.93) were observed for transforming growth factor beta 3 (TGFbeta3) in HLA DR15-associated families, suggesting a possible role for this growth factor in interaction with HLA. CONCLUSIONS: Oligodendrocyte growth factors do not play a significant role in MS genetic susceptibility, at least in the tested sample. TGFbeta3, the only gene highlighted by this study, deserves further analysis. PMID- 9748022 TI - Increased release of interleukin-12p40 in MS: association with intracerebral inflammation. AB - OBJECTIVE: The p40 subunit of interleukin (IL)-12 was recently demonstrated in active lesions in MS. We tested whether the p40 subunit of IL-12 can also be detected in CSF and serum of patients with this disease and, if so, whether release is associated with inflammatory disease activity. RESULTS: This study demonstrates an increased (up to 1,000-fold) compartmentalized release of the p40 subunit but not of the heterodimer p70 in MS. Release of IL-12p40 correlated with classic markers of CNS inflammation (CSF cell counts, immunoglobulin G index) and was significantly increased in patients with gadolinium-enhancing plaques on MRI. Moreover, release of IL-12p40 was associated with CSF levels of myelin basic protein as a measure of myelin degradation. CONCLUSION: These results suggest a role of IL-12p40 in the pathophysiology of MS. PMID- 9748023 TI - Serial magnetization transfer imaging to characterize the early evolution of new MS lesions. AB - OBJECTIVE: To explore the temporal relation of demyelination and blood-brain barrier breakdown during new lesion formation. BACKGROUND: Conventional MRI appears sensitive for detecting changes due to MS, but may be limited by poor pathologic specificity. By indirectly assessing protons bound to rigid macromolecules, magnetization transfer (MT) imaging may provide information relating to tissue structure and, by inference, myelin integrity. METHODS: Gadolinium contrast-enhanced MRI and MT imaging were performed at weekly intervals for 3 months in three patients with MS. For each enhancing lesion, the largest corresponding area of proton density hyperintensity seen during the study was outlined and magnetization transfer ratio (MTR) calculated at each time point from coregistered calculated MTR images. Lesions greater than 20 mm2, not affected by partial volume effects, and first enhancing after the baseline study were analyzed. Two-dimensional registration software allowed accurate evaluation of MTR in regions both before and after the initial appearance of MS lesions. RESULTS: Mean lesion MTR decreased significantly during the first week of enhancement (29.6 percent units [pu] immediately pre-enhancement versus 28.2 pu at first documented stage of enhancement). No significant MTR reduction was noted before this. CONCLUSION: The lack of observable change in MTR before the first detectable gadolinium enhancement within MS lesions suggests that blood-brain barrier disruption is closely related to, but not preceded by, demyelination. PMID- 9748024 TI - Survival and predictors of disability in Turkish MS patients. Turkish Multiple Sclerosis Study Group (TUMSSG). AB - OBJECTIVE: To examine the natural history, survival, and prognostic factors in a sample of Turkish MS patients. METHOD: This multicenter study included 1,259 definite MS patients diagnosed according to the criteria of Poser et al. Actuarial analysis of selected disability levels of 3, 6, 8, and 10 achieved with the Expanded Disability Status Scale (EDSS); a multivariate Cox regression analysis for prognostic factors related to time to reach EDSS > or = 6; and Pearson's correlation coefficient for individual factors were performed. RESULTS: The survival (+/- SE) at 15 years from onset was 94.6 +/- 2.9%, and at 25 years was 89.0 +/- 5.8%. The disability reached by 15 years was EDSS > or = 3 in 66.4%, EDSS > or = 6 in 41.2%, EDSS > or = 8 in 10.5%, and EDSS = 10 in 5.4%. The most significant unfavorable prognostic factors were progressive course (relative risk [RR], 3.73; CI, 2.71 to 5.13) and sphincter symptoms at onset (RR, 1.86; CI, 1.23 to 2.82), followed by male sex, motor symptoms at onset, and a high attack frequency within the first 5 years. Primary progressive disease was correlated positively with male sex (r = 0.0895, p = 0.001), older age (r = 0.1807, p = 0.000), and motor (r = 0.1433, p = 0.000) or sphincter symptoms (r = 0.1001, p = 0.000) at onset, unlike relapsing-remitting and secondary progressive disease. CONCLUSIONS: Although a slightly better prognosis is observed in the Turkish MS population, early prognostic factors are similar to most of the previous Western series. Primary progressive disease, mostly seen in older men with motor and sphincter involvement at onset, has a worse prognosis and may represent a distinct behavioral variant of MS. PMID- 9748025 TI - A placebo-controlled crossover study of rizatriptan in the treatment of multiple migraine attacks. Rizatriptan Multiple Attack Study Group. AB - OBJECTIVE: To examine the safety and efficacy of rizatriptan 10 mg PO in the treatment of multiple migraine attacks. BACKGROUND: Rizatriptan is a potent and rapidly absorbed 5-HT1B/1D receptor agonist. Efficacy and general safety have been examined in controlled trials treating single migraine attacks. In the current placebo-controlled study, we report constancy of safety and efficacy of rizatriptan for patients treating four discrete migraine attacks. METHODS: Patients with moderate or severe migraine (n = 473) were randomized to one of five sequence groups, in which each patient was to treat four migraine attacks. Patients in four groups received rizatriptan 10 mg for three of four attacks and placebo for the remaining attack. Patients in the fifth group received rizatriptan 10 mg for four attacks. Headache severity, functional disability, and migraine symptoms were measured immediately before dosing and at 0.5, 1, 1.5, 2, 3, and 4 hours postdose. RESULTS: After the first attack, response rates were 77% for rizatriptan and 37% for placebo (p < 0.001). Similar efficacy of rizatriptan, ranging from a 75 to 80% response, was observed in each of the subsequent attacks with no evidence of tolerance to therapeutic effects. Most patients (93%) responded to rizatriptan 10 mg during the first or second attack. Adverse experiences were generally mild and transient, the most common being dizziness and somnolence. Incidence of adverse experiences per attack decreased after the first attack. CONCLUSIONS: Rizatriptan 10 mg PO is efficacious and generally well tolerated in acute migraine. Its efficacy is maintained throughout the treatment of multiple, discrete migraine attacks. PMID- 9748026 TI - Association between dopamine receptor genes and migraine without aura in a Sardinian sample. AB - BACKGROUND: Migraine seems to be caused by a combination of environmental and genetic factors. Clinical and pharmacologic evidence supports the hypothesis that dopaminergic transmission is involved in the pathogenesis of migraine. OBJECTIVE: The current report concerns a genetic study to test the involvement of genes for dopamine (DA) receptors D2 (DRD2), D3 (DRD3), and D4 (DRD4) in migraine without aura, particularly in a subgroup with enhanced DA sensitivity. METHODS: For the first time, a family-based association method--the Transmission Disequilibrium Test (TDT)--was used to examine an isolated population, such as Sardinians. We studied 50 nuclear families of patients affected by migraine without aura. The subgroup of dopaminergic migraineurs was selected based on the presence of both nausea and yawning immediately before or during the pain phase of migraine. RESULTS: No association was detected using the TDT between DRD3, DRD4, and migraine without aura either in the overall sample or in the subgroup. No difference was observed in DRD2 allelic distribution in the overall sample, although the allelic distribution at the DRD2 locus differed significantly in the subgroup of dopaminergic migraineurs (p = 0.004). Allele 1 of the TG dinucleotide intronic noncoding polymorphism of the DRD2 locus was the individual allele that appeared to be in disequilibrium with migraine without aura (p = 0.02). CONCLUSIONS: Our data suggest that a genetic approach could be useful in providing molecular support to the hypothesis that hypersensitivity of the dopaminergic system may represent the pathophysiologic basis of migraine, at least in a subgroup of patients. PMID- 9748027 TI - Orthostatic headaches caused by CSF leak but with normal CSF pressures. AB - OBJECTIVE: To report that the syndrome of orthostatic headaches caused by CSF leak can be seen with persistently normal CSF pressures. BACKGROUND: CSF leak or shunt overdrainage is known to cause orthostatic headaches and diffuse pachymeningeal gadolinium enhancement (DPGE), typically associated with unmeasurable or very low CSF pressures. METHODS: Of 40 consecutive patients with orthostatic headaches and DPGE, all had low or unmeasurable CSF pressures, except seven patients who had consistently normal CSF pressures and are thus reported. All had undergone multiple CSF examinations. RESULTS: Two patients had overdraining shunts, and five had documented CSF leaks. One refused treatment, but the other six patients responded to surgical treatment or epidural blood patch with complete resolution of symptoms and related MRI abnormalities. CONCLUSIONS: Some patients with symptomatic CSF leaks may have CSF opening pressures that are consistently within normal limits. In the presence of convincing clinical features and imaging abnormalities, a normal CSF pressure should not discourage the clinician from searching for a source of CSF leak. PMID- 9748028 TI - Chronic traumatic brain injury in professional soccer players. AB - OBJECTIVE: To determine the presence of chronic traumatic brain injury in professional soccer players. METHODS: Fifty-three active professional soccer players from several professional Dutch soccer clubs were compared with a control group of 27 elite noncontact sport athletes. All participants underwent neuropsychological examination. The main outcome measures were neuropsychological tests proven to be sensitive to cognitive changes incurred during contact and collision sports. RESULTS: The professional soccer players exhibited impaired performances in memory, planning, and visuoperceptual processing when compared with control subjects. Among professional soccer players, performance on memory, planning, and visuoperceptual tasks were inversely related to the number of concussions incurred in soccer and the frequency of "heading" the ball. Performance on neuropsychological testing also varied according to field position, with forward and defensive players exhibiting more impairment. CONCLUSION: Participation in professional soccer may affect adversely some aspects of cognitive functioning (i.e., memory, planning, and visuoperceptual processing). PMID- 9748029 TI - Pallidotomy and bradykinesia: implications for basal ganglia function. AB - BACKGROUND AND OBJECTIVE: The scientific rationale for pallidotomy as a treatment for PD is that the lesion will reduce excessive tonic inhibition of the thalamus, thereby allowing movement to proceed more normally. If true, then PD patients who move slowly while on medication should increase movement speed following pallidotomy. To test this we used a simple motor task to determine if pallidotomy leads to an improvement in "on" motor performance when those movements are impaired before surgery. METHODS: Nine patients with PD performed elbow flexion movements "as fast as possible" while they were "on" before and 1 month after pallidotomy. Patients with mild PD and healthy control subjects were also tested. RESULTS: The clinical effects of pallidotomy were typical of those found in other studies. "Off" Unified Parkinson's Disease Rating Scale scores improved and dyskinesias were reduced. Although before surgery the patients were far slower while they were "on" than the groups of mild PD patients and healthy control subjects, there was no change in mean peak velocity while they were "on" after pallidotomy. There was no change in other mean "on" motor performance measures such as peak acceleration, peak deceleration, initiation time, and symmetry. There was a decrease in the variability of peak acceleration, symmetry, and initiation time. CONCLUSION: Despite the clinical efficacy of pallidotomy while patients were "off," bradykinesia of elbow flexion movements while patients were "on" is not affected by pallidotomy. Therefore, we conclude that the bradykinesia observed in this experiment is due to a mechanism other than excessive tonic inhibition of the motor thalamus. Our results are consistent with the idea that pallidotomy reduces the noise from the abnormally functioning basal ganglia. PMID- 9748030 TI - The metabolic anatomy of tremor in Parkinson's disease. AB - OBJECTIVE: To identify regional metabolic brain networks related specifically to the presence of tremor in PD. BACKGROUND: The pathophysiology of parkinsonian tremor is unknown. Because tremor in PD occurs mainly in repose, we used resting state PET with 18F-fluorodeoxyglucose (FDG) to identify specific metabolic brain networks associated with this clinical manifestation. METHODS: We studied two discrete groups of eight PD patients with and without tremor using FDG/PET. Both patient groups were matched for gender, age, and Unified Parkinson Disease Rating Scale ratings for akinesia and rigidity. Ten normal volunteer subjects served as controls. RESULTS: Network analysis with the Scaled Subprofile Model was performed in two steps. 1) We computed the expression of the PD-related pattern (PDRP) identified by us previously in each of the PD patients and control subjects. Although PDRP subject scores were abnormally elevated in the combined PD cohort (p < 0.005), these values did not differ in the PD patient groups with and without tremor (p = 0.36). 2) We used SSM to analyze the data from the combined PD cohort comprising both patient groups. We found that PD patients with tremor were characterized by increased expression of a metabolic network comprising the thalamus, pons, and premotor cortical regions. Subject scores for this pattern were elevated in the tremor group compared with the atremulous patient group and the normal control group (p < 0.005). CONCLUSIONS: The findings suggest that PD patients with tremor are characterized by distinct increases in the functional activity of thalamo-motor cortical projections. Modulation of this functional anatomic pathway is likely to be the mechanism for successful interventions for the relief of parkinsonian tremor. PMID- 9748031 TI - Early dopaminergic drug-induced hallucinations in parkinsonian patients. AB - OBJECTIVE: To characterize patients who develop hallucinations early in the course of dopaminergic therapy for Parkinson's disease (PD) and contrast them with patients developing hallucinations after chronic drug treatment. METHODS: Parkinsonian patients who met diagnostic criteria for PD, experienced hallucinations, had a detailed hallucination interview at the onset time of their first hallucination, and had a 5-year clinical follow-up or an autopsy in those 5 years were identified and divided into two groups for comparison: 12 patients who developed early hallucinations within 3 months of starting levodopa therapy and 58 PD patients who developed hallucinations after 1 year of dopaminergic therapy. We contrasted the quality, content, diurnal nature, and emotional elements of the hallucinations, as well as the 5-year follow-up data on diagnosis, disease course, community home or nursing home outcome, and mortality. RESULTS: Both groups experienced a predominance of visual hallucinations, visions of people and animals, and vivid colors and definition. Features distinctive to the early onset hallucinating patients included visions that persisted in daytime as well as nighttime, frightening content with paranoia, and accompanying nonvisual hallucinations, either auditory, olfactory, tactile, or combinations thereof. At the 5-year follow-up, none of the early onset hallucinators had PD as their sole disorder. Four of the 12 had an underlying psychiatric illness that included hallucinations or psychosis preceding their parkinsonism by several years. In the other eight patients at the 5-year follow-up, their parkinsonism evolved to include additional signs that were no longer consistent with PD. The primary diagnoses were diffuse Lewy body disease and Alzheimer's disease (AD) with extrapyramidal signs. Patients with early drug-induced hallucinations had significantly greater placement to nursing homes and greater mortality. CONCLUSIONS: Early onset drug-related hallucinations are not typical of PD. Their presence should signal an investigation of two alternative diagnoses, either a comorbid psychotic illness (often unrevealed by the patient initially) or an evolving parkinsonism-plus syndrome. PMID- 9748032 TI - Electromyography in cervical dystonia: changes after botulinum and trihexyphenidyl. AB - BACKGROUND: The value of physical examination in detecting involved neck muscles in cervical dystonia (CD) is uncertain and little is known about changes in electromyographic (EMG) features after botulinum toxin type A (BTA) treatment. METHODS: In a double-blind, randomized study we recorded the EMG activities of 420 neck muscles in 42 patients with CD before and after treatment with BTA or trihexyphenidyl. We regarded any needle EMG activity higher than 100 microV as the gold standard for involuntary involvement of a muscle in the dystonic posture and compared this with the results of physical examination. We calculated EMG total scores by adding the scores of the individual muscles. RESULTS: Physical examination had a low predictive value in the detection of involved muscles. There was a significant correlation between changes in EMG total scores and changes in clinical measurements. We observed increased EMG activity in 20% of noninjected muscles after BTA treatment and in 27% of noninjected muscles after trihexyphenidyl treatment. A switch from one most active muscle to another was seen equally in both groups and had no influence on clinical response. CONCLUSION: Physical examination alone is not sufficient to detect involved muscles, and repeated, simultaneous EMG-guided application of BTA may be helpful. In addition to clinical measurements, changes in EMG activity due to treatment can be used as a physiologic measure in evaluating treatment response. Increased activity of noninjected muscles and a switch from one most active muscle to another are not related to BTA treatment, but are probably pathophysiologic phenomena of CD itself. PMID- 9748033 TI - Putamen volume in idiopathic focal dystonia. AB - OBJECTIVE: To determine whether the volume of the putamen is abnormal in patients with idiopathic focal dystonia. BACKGROUND: The cause of adult-onset focal dystonia is unknown, but substantial evidence suggests that the putamen may be abnormal in this condition. Cell loss and gliosis have been suggested. We hypothesized that this might be reflected as abnormal putamen volume on MRI. DESIGN AND METHODS: A high-resolution MRI was acquired in 13 adults with cranial or hand dystonia and 13 normal individuals matched for age and sex. Putamen volume was measured using a stereologic method (Study 1). In a replication study, another rater measured putamen volume using manual tracing and direct voxel count (Study 2). Neither rater was aware of the diagnosis, and the order of measurement was random in each study. RESULTS: In Study 1, putamen measurements were reasonably accurate (coefficient of error, approximately 6%). The putamen was 13% larger in patients, both in absolute terms (p = 0.03) and after covarying total brain volume (p = 0.02). In Study 2, putamen volumes correlated with those measured in Study 1 (intraclass correlation coefficient, 0.68 to 0.83). The putamen was 8% larger in patients (p = 0.06) and was larger in the patient than in the matched control subject in 10 of 13 pairs (p = 0.046). CONCLUSION: We find no evidence of putaminal atrophy or degeneration in adult-onset idiopathic focal dystonia. In fact, in this group, the putamen is about 10% larger in patients than in matched control subjects. This finding may reflect a response to the dystonia or may relate to its cause. PMID- 9748034 TI - Effect of selegiline on mortality in patients with Parkinson's disease: a meta analysis. AB - INTRODUCTION: The Parkinson's Disease Research Group of the United Kingdom (PDRG UK) reported increased mortality in PD patients treated with levodopa plus selegiline compared with those treated with levodopa alone. METHODS: We performed a meta-analysis on five long-term, prospective, randomized trials of selegiline in patients with untreated PD. Included in the analysis were four randomized, double-blind, placebo-controlled studies and one randomized, double-blind, placebo-controlled study of 2 years' duration followed by long-term, open follow up. RESULTS: The mean duration of follow-up was 4.1 +/- 1.8 years. There were 14 deaths in 297 selegiline-treated patients (4.7%) and 17 deaths in 292 non selegiline-treated patients (5.8%). The hazard ratio for mortality was 1.02 (95% CI 0.44 to 2.37; p = 0.96). An analysis restricted to patients receiving only levodopa with or without selegiline noted 11 deaths in 257 levodopa/selegiline treated patients (4.3%) and 11 deaths in 254 patients treated with levodopa alone (4.3%). The hazard ratio was 1.06 (95% CI 0.44 to 2.55; p = 0.90). Death rate per 1,000 patient years was 11.4 in the selegiline group and 14.2 in the nonselegiline group. Kaplan-Meier survival curves reflecting pooled survival data showed no significant difference in duration of survival. The hazard ratio was 0.84 (95% CI 0.41 to 1.70; p = 0.63) for selegiline- versus non-selegiline treated patients and 1.05 (95% CI 0.46 to 2.43; p = 0.91) for selegiline/levodopa versus levodopa-treated patients. CONCLUSION: These results contrast with those of the PDRG-UK study and demonstrate no increase in mortality associated with selegiline treatment whether or not patients also received levodopa. PMID- 9748035 TI - Seesaw nystagmus associated with involuntary torsional head oscillations. AB - OBJECTIVE: To assess the diagnostic value of eye-head coupling in seesaw nystagmus (SSN). BACKGROUND: SSN is a rare binocular disorder characterized by alternating skew deviation and conjugate ocular torsion. METHODS: We examined a patient with a congenital nystagmus that switched to a pendular SSN on near viewing and was associated with involuntary torsional head oscillations. RESULTS: The binocular torsional eye movements were in phase with the clinically visible head oscillations (i.e., head movements were not compensatory for the torsional eye movements). CONCLUSION: This finding suggests that torsional eye-head coupling in pendular SSN has a common pathologic origin. We suggest that alternating vertical disparity of both eyes in pendular SSN is compatible with an oscillating signal acting on an intact vestibular system. The absence of brainstem lesions on high-resolution MRI supports this assumption. PMID- 9748036 TI - Vestibular exercises improve central vestibulospinal compensation after vestibular neuritis. AB - OBJECTIVE AND BACKGROUND: Animal experiments have shown that central vestibular compensation of unilateral peripheral vestibular lesions can be improved by vestibular exercises. There are, however, no equivalent clinical studies on the efficacy of such specific physiotherapy on acute unilateral peripheral vestibular lesions in humans. DESIGN AND METHODS: To quantify the differential effects of specific vestibular exercises on central compensation in patients with an acute/subacute unilateral vestibular lesion (vestibular neuritis), we determined the time course of recovery of 1) the ocular torsion (OT) for the vestibulo ocular system, 2) the subjective visual vertical (SVV) for perception, and 3) the total sway path (SP) values for postural control in 19 patients with and 20 patients without vestibular exercises. All patients had a persisting peripheral vestibular deficit for at least 30 days (statistical end point). RESULTS: Although normalization of OT and SVV was similar in the control and physiotherapy groups, the total SP values on day 30 after symptom onset differed significantly: 3.2 +/- 1.9 m/min in the physiotherapy group and 16.9 +/- 6.1 m/min in the control group (ANOVA, p < 0.001). CONCLUSIONS: This prospective clinical study suggests that specific vestibular exercises improve vestibulospinal compensation in patients with acute peripheral vestibular lesions. PMID- 9748037 TI - CADASIL in a North American family: clinical, pathologic, and radiologic findings. AB - OBJECTIVE: To expand the reported phenotypic range of cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL). BACKGROUND: Despite numerous patient reports, our knowledge of the phenotypic range of CADASIL remains incomplete. METHOD: We performed clinical, pathologic, and radiologic examinations on members of a family with CADASIL. RESULTS: The proband is a 61-year-old man with a history of migraine and depression who has experienced multiple subcortical infarctions resulting in a stepwise decline. Neuropsychological testing documented a dementia syndrome with frontal lobe features and neurologic examination noted a left hemiparesis and a right-sided palmomental reflex. Brain biopsy with light microscopy revealed a nonatherosclerotic small-vessel angiopathy with periodic acid-Schiff positive granular changes in the media and white matter gliosis, with unremarkable cortex. Genetic testing confirmed a Notch3 mutation. The proband's first cousin has a history of depression, one seizure possibly resulting from an acute stroke, and a learning disorder. Neuropsychological testing demonstrated deficits in executive function and neurologic examination noted persistent extraneous adventitial movements, poor coordination, and primitive reflexes. Skin biopsy with electron microscopy demonstrated granular osmiophilic material within the basement membrane of vascular smooth muscle cells, which is considered to be pathognomonic of CADASIL. The proband's older son and younger son have histories of migraine and depression, respectively, and both also had learning disorders. MRI revealed diffuse white matter disease extending into the temporal lobes, and lacunar infarctions in these four nonhypertensive patients. Other family members have experienced migraine, recurrent stroke, dementia, and depression. CONCLUSIONS: CADASIL is a genetic basis for vascular dementia that may be manifest earlier in life than previously reported. PMID- 9748038 TI - Double-blind evaluation of subthalamic nucleus deep brain stimulation in advanced Parkinson's disease. AB - OBJECTIVE: To examine objectively the clinical effects of subthalamic nucleus (STN) deep brain stimulation (DBS) in advanced PD. METHODS: Our initial seven consecutive patients with medication-refractory motor fluctuations and levodopa induced dyskinesias undergoing chronic STN DBS underwent a standardized preoperative evaluation followed by a 2-day double-blind evaluation of efficacy 6 to 12 months after electrode implantation. Diaries documenting motor fluctuations and dyskinesias were also completed preoperatively and postoperatively. RESULTS: In the medication-off state, turning the stimulators on resulted in improvement in mean total Unified Parkinson's Disease Rating Scale (UPDRS) motor score by 58% including the following improvements in composite scores: akinesia 57%, rigidity 52%, tremor 82%, and gait and postural stability 49%. Additionally, the medication-off state improved 17% without stimulation, possibly as a result of electrode insertion alone or carry-over of chronic stimulation. In the medication on, stimulation-on state, all major features of parkinsonism improved and total UPDRS motor score improved 41% compared with before surgery. Activities of daily living were improved while off medication 30%, and levodopa-induced dyskinesias were reduced 83% while total drug dosage was decreased 40%. With chronic stimulation, patients reported that the percentage of time spent in the "on" state (without dyskinesias) increased from 26% to 52% and "off" time decreased from 30% to 6%. Operative complications including cognitive worsening were not uncommon. CONCLUSIONS: STN DBS is a promising new surgical option for the treatment of advanced PD. The marked clinical benefits obtained in these severely disabled patients outweighed the adverse effects. PMID- 9748039 TI - Rabies: a possible explanation for the vampire legend. AB - In the 18th century, belief in vampires--allegedly dead persons who left their graves and killed people and animals--raised great concern in the Balkans and an extensive debate in Europe. This historic phenomenon still awaits a comprehensive explanation. This article proposes that rabies may have played a key role in the development of the vampire legend, given the coincident time of the phenomena and the striking similarities between them. PMID- 9748040 TI - Ahomocysteinemia in molybdenum cofactor deficiency. AB - We report an infant with molybdenum cofactor deficiency (MCD) and a unique clinical presentation of hemiplegia, hypotonia, dystonia, and bilateral basal ganglia changes. Biochemistry revealed absent serum homocysteine, low concentrations of plasma cystine, high levels of urinary S-sulfocysteine and sulfite, and high levels of oxypurines in serum and urine. The depletion of cysteine and cystine through reaction with sulfite suggests that other thiols and thiol-dependent proteins may be similarly depleted. Ahomocysteinemia may be a clue to the mechanism of cytotoxicity in MCD. PMID- 9748041 TI - Cerebral manifestation of Wilson's disease successfully treated with liver transplantation. AB - The main indication for orthotopic liver transplantation (OLTx) in Wilson's disease (WD) is severe hepatic decompensation. Our 15-year-old patient is the second case to date in whom OLTx was performed because of neurologic manifestations resulting from WD. His initial condition involving recurrent headaches, tremor, and athetoid hand movements progressively deteriorated during therapy with D-penicillamine, zinc sulfate, and trientine until he was severely dysarthric, unable to walk, and bedridden. After OLTx, his neurologic condition became almost normal. PMID- 9748042 TI - Genetic analysis enables definite and rapid diagnosis of cerebrotendinous xanthomatosis. AB - Mutations in the sterol 27-hydroxylase gene (CYP27) cause cerebrotendinous xanthomatosis (CTX). Early diagnosis of CTX is crucial because treatment with chenodeoxycholic acid can prevent or reverse some of the neurologic disability associated with the disease. We report the identification of three types of mutations (Arg441Trp, Arg372Gln, and Arg441Gln) in the CYP27 gene in five patients with suspected CTX from four unrelated families by restriction endonuclease analysis. PMID- 9748043 TI - Reversible peripheral neuropathy induced by a single administration of high-dose paclitaxel. AB - Peripheral neuropathy (PN) is the main side effect with cycles of paclitaxel at standard doses (175 mg/m2 for 21 days). Administration of a single high-dose paclitaxel (HDP) is a novel approach for the treatment of cancer. We have prospectively measured neurotoxicity induced by HDP during a phase I trial. Nineteen patients were treated with escalating doses of paclitaxel by 24-hour infusion. In our study, PN induced by HDP was moderate, reversible, and not dose limiting. Severe PN was seen in patients who had received previous neurotoxic chemotherapy, and caution on the administration of HDP in this setting is warranted. PMID- 9748045 TI - Congenital cytoplasmic body myopathy with survival motor neuron gene deletion or Werdnig-Hoffmann disease. AB - A 5-week-old boy became rigid and developed cardiac arrest after receiving succinylcholine. He was resuscitated and ventilated but died at 5 months. Muscle biopsy demonstrated no neurogenic features and numerous cytoplasmic bodies, suggesting the possibility of congenital myopathy with cytoplasmic bodies. However, molecular analysis revealed a homozygous deletion of exons 7 and 8 of the survival motor neuron (SMN) gene, suggesting that the patient had Werdnig Hoffmann disease. We recommend that every patient with congenital cytoplasmic body myopathy be tested for SMN gene deletion. PMID- 9748044 TI - Familial amyotrophic lateral sclerosis with widespread vacuolation and hyaline inclusions. AB - This report presents a familial amyotrophic lateral sclerosis (FALS) patient with widespread vacuoles and hyaline inclusions strongly immunostained with the anti superoxide dismutase (SOD1) antibody. The overall pathologic similarity between our non-SOD1-linked FALS patient and transgenic mice expressing a mutated human SOD1 gene suggests that common pathogenetic mechanisms other than an SOD1 mutation exist in the development of these diseases. PMID- 9748046 TI - Minimal number of plasma exchanges needed to reduce immunoglobulin in Guillain Barre syndrome. AB - Plasma exchange (PE) in Guillain-Barre syndrome (GBS) probably removes pathogenic antibodies. Results of a recent clinical study by the French Cooperative Group suggest that at least two sessions of PE are required. As an alternative procedure, we examined the effect of the number of PEs on the reduction of immunoglobulins in 11 patients. A significant immunoglobulin decrease was obtained in the first two sessions but not in subsequent ones. Based on the French trial results and our findings, we conclude that at least two PEs are needed for treating GBS. PMID- 9748047 TI - Large-scale deletions in a Chinese infant associated with a variant form of Werdnig-Hoffmann disease. AB - A Chinese male infant with arthrogryposis multiplex congenita (AMC), ventricular and atrial septal defects, and Werdnig-Hoffmann disease (WHD) had deletions of the telomeric copy of the survival motor neuron (SMN(T)) and neuronal apoptosis inhibitory protein genes. Children with AMC or congenital heart disease, or both, and motor neuron disease should undergo testing for SMN(T) deletion. This rare association further illustrates the variable phenotypic expressions of WHD. PMID- 9748048 TI - Gabapentin in orthostatic tremor: results of a double-blind crossover with placebo in four patients. AB - We treated four patients affected by orthostatic tremor (OT) with gabapentin in increasing doses (300 to 2,400 mg/d). OT was evaluated with patients' self monitoring scales, tremor rating scales, electromyography (EMG) showing the 14- to 18-Hz frequencies, and EMG frequency analysis. All patients had transitory responses to clonazepam. Gabapentin induced disappearance of OT in three patients and consistent reduction in one. Crossover to placebo induced reappearance of tremor. PMID- 9748049 TI - CAG repeat number correlates with the rate of brainstem and cerebellar atrophy in Machado-Joseph disease. AB - We compared the CAG repeat length and the severity of the brainstem and cerebellar atrophy visualized by MRI in 30 patients with Machado-Joseph disease. We found a strong correlation between the CAG repeat number and the quotient of the degree of atrophy divided by age at examination. These results suggest that the rate of disease progression is dependent on the CAG repeat size and disease progression may commence at birth. PMID- 9748050 TI - The predictive value of CSF oligoclonal banding for MS 5 years after optic neuritis. Optic Neuritis Study Group. AB - The predictive value of CSF oligoclonal banding for the development of clinically definite MS (CDMS) within 5 years after optic neuritis was assessed in 76 patients enrolled in the Optic Neuritis Treatment Trial. The presence of oligoclonal bands was associated with the development of CDMS (p = 0.02). However, the results suggest that CSF analysis is only useful in the risk assessment of optic neuritis patients when brain MRI is normal and is not of predictive value when brain MRI lesions are present at the time of optic neuritis. PMID- 9748051 TI - Lewy body in neurodegeneration with brain iron accumulation type 1 is immunoreactive for alpha-synuclein. AB - In familial PD, a mutation of the alpha-synuclein gene has been identified. Alpha synuclein also was revealed in Lewy bodies in idiopathic PD. Lewy bodies in neurodegeneration with brain iron accumulation type 1 (NBIA 1; Hallervorden-Spatz syndrome) were found to show immunostaining for alpha-synuclein/precursor of non A beta component of Alzheimer's disease amyloid, indicating that alpha-synuclein is commonly associated with the formation of Lewy bodies in other sporadic and familial neurodegenerative diseases apart from PD. PMID- 9748052 TI - Pathologic and biochemical studies of juvenile parkinsonism linked to chromosome 6q. AB - We report the results of pathologic and biochemical studies in a patient with 6q linked autosomal recessive juvenile parkinsonism (AR-JP). Neuronal loss and gliosis were restricted to the substantia nigra and the locus ceruleus. No Lewy bodies were found, but neurofibrillary tangles and argyrophilic astrocytes were seen in the cerebral cortex and brainstem nuclei. The later findings, which have not been reported previously in AR-JP, suggest the pathologic heterogeneity of 6q linked AR-JP. PMID- 9748053 TI - Proton magnetic resonance spectroscopy in primary blepharospasm. AB - Single-volume proton magnetic resonance spectroscopy, localized to basal ganglia, was carried out in 10 patients with primary blepharospasm (PB) to assess the levels of N-acetyl aspartate (NAA), creatine-phosphocreatine, and choline containing compounds. NAA was reduced significantly in patients compared with control subjects. This result suggests a striatal neuronal loss in PB. PMID- 9748054 TI - Effects of the serotonin1B/1D receptor agonist zolmitriptan on motor cortical excitability in humans. AB - Oral administration of zolmitriptan, a novel 5-hydroxytriptamine receptor agonist, to eight healthy volunteers significantly reduced motor cortical excitability as tested by paired transcranial magnetic stimulation (TMS) at short interstimulus intervals. Zolmitriptan did not change motor thresholds to TMS or electromyographic silent period durations. We conclude that zolmitriptan acts centrally by reducing the inhibition within the motor cortex. The results suggest that the action of zolmitriptan on motor cortical excitability does not result from changes at the level of the cell membrane but from the influence on GABAergic inhibitory interneurons. PMID- 9748055 TI - Brain infarction following 5-fluorouracil and cisplatin therapy. AB - Five patients with oropharyngeal cancer treated with 5-fluorouracil and cisplatin had ischemic stroke within 2 to 5 days after the drug infusion. This occurred during the second course of chemotherapy in three patients, and during the third course in two patients. There may be a relation between treatment and brain infarction because 1) there was no other cause identified despite extensive tests, including postmortem examination in one patient; 2) there was a short delay between treatment infusion and stroke; and 3) there was a similar pattern of ischemic stroke after the second or third course of chemotherapy. PMID- 9748056 TI - Diffusion-weighted MRI characterizes the ischemic lesion in transient global amnesia. AB - We present a patient with transient global amnesia (TGA) whose diffusion-weighted MRI (DWI) showed increased signal in the splenium of the corpus callosum and in the left parahippocampal gyrus. The absence of high signal on the corresponding apparent diffusion coefficient (ADC) images supports the diagnosis of an acute infarction. This finding provides a temporal relation between cerebral ischemia and infarction in the territory of posterior cerebral artery and in certain cases of TGA. An early means of detecting ischemia in TGA by DWI may influence clinical decisions made in patient evaluation and management. PMID- 9748057 TI - A maternal complex partial seizure in labor can affect fetal heart rate. AB - We describe a 33-year-old woman who had a complex partial seizure during labor. Intrauterine pressure catheter and fetal heart monitoring during the seizure revealed a strong, prolonged uterine contraction and simultaneous significant fetal heart rate deceleration for 3.5 minutes. This patient demonstrates that complex partial seizures may result in uterine hyperactivity during labor, which may result in fetal hypoxia. PMID- 9748058 TI - The value of interphase cytogenetics in cytology for the diagnosis of leptomeningeal metastases. AB - We studied the use of fluorescence in situ hybridization (FISH) in CSF to enhance the diagnostic yield for the detection of malignancy on the first lumbar puncture in patients clinically suspected of having leptomeningeal metastases (LMM). Although repeated lumbar punctures were still needed in some patients, the use of FISH did speed up the diagnosis in approximately one-third of the patients clinically suspected of having LMM with atypical cells at first cytology. This eliminates the need for repeated lumbar punctures in these patients and enables an earlier start of treatment. PMID- 9748059 TI - Acute therapy for cluster headache with sumatriptan: findings of a one-year long term study. AB - The efficacy, safety, and tolerability of subcutaneous sumatriptan in the acute treatment of cluster headache were investigated in a multicenter study over a period of up to 1 year. A total of 2,031 attacks were evaluated in 52 patients. Therapy was successful in 88% of all attacks. Freedom from pain within 15 minutes in more than 90% of all attacks treated was reported by 42% of the patients, and no decline in efficacy occurred during the course of the study. Adverse events were reported by 62% of the patients. PMID- 9748060 TI - The importance of formal serum iron studies in the assessment of restless legs syndrome. PMID- 9748061 TI - Successful treatment of a patient with severe refractory myasthenia gravis using mycophenolate mofetil. PMID- 9748062 TI - Myasthenic hand. PMID- 9748063 TI - Varicella zoster virus-associated focal vasculitis without herpes zoster: recovery after treatment with acyclovir. PMID- 9748064 TI - Reversible occipital-parietal encephalopathy syndrome in an HIV-infected child. PMID- 9748065 TI - Basilar artery occlusion associated with pathological crying: 'folles larmes prodromiques'? PMID- 9748066 TI - Amiodarone-induced severe prolonged head-positional vertigo and vomiting. PMID- 9748067 TI - Steroid myopathy in cancer patients. PMID- 9748068 TI - Electroencephalography and survival in patients with Alzheimer's disease. PMID- 9748069 TI - Disruption of short-duration timing associated with damage to the suprachiasmatic region of the hypothalamus. PMID- 9748070 TI - Lesion laterality, neglect, and anosognosia. PMID- 9748071 TI - Inaccuracy of the ICD-9-CM in identifying the diagnosis of ischemic cerebrovascular disease. PMID- 9748072 TI - Inaccuracy of the ICD-9-CM in identifying the diagnosis of ischemic cerebrovascular disease. PMID- 9748073 TI - Inaccuracy of the ICD-9-CM in identifying the diagnosis of ischemic cerebrovascular disease. PMID- 9748074 TI - Quality improvement in neurology residency programs. PMID- 9748075 TI - Orthostatic tremor associated with a pontine lesion or cerebellar disease. PMID- 9748076 TI - Reliability of the NINDS Myotatic Reflex Scale. PMID- 9748077 TI - Pain and opioid analgesics in Guillain-Barre syndrome. PMID- 9748078 TI - Recurrent hemorrhage in cerebral amyloid angiopathy. PMID- 9748079 TI - Dopamine and migraine. PMID- 9748080 TI - Insulin, diabetes mellitus, Alzheimer's disease, and apolipoprotein E. PMID- 9748081 TI - Improvement in the polyneuropathy associated with familial amyloid polyneuropathy after liver transplantation. PMID- 9748082 TI - Pain after thalamic stroke: right diencephalic predominance and clinical features in 180 patients. PMID- 9748083 TI - Pain after thalamic stroke: right diencephalic predominance and clinical features in 180 patients. PMID- 9748084 TI - The D2 receptor NcoI allele: absence of allelic association with migraine with aura. PMID- 9748085 TI - Exercise-induced increase in core temperature does not disrupt a behavioral measure of sleep. AB - On separate nights 90 to 30 min before typical bedtime, eight physically active men completed three conditions: seated rest, low-intensity and moderate-intensity cycle exercise. Low-and moderate-intensity exercise had no significant effect on sleep onset latency, the number of awakenings, total sleep time or sleep efficiency as measured by the Sleep Assessment Device. Mean core body temperature was higher during sleep after moderate intensity (36.80+/-0.02 degrees C) exercise compared to both the low-intensity exercise (36.67+/-0.02 degrees C) and rest (36.51+/-0.02 degrees C) conditions. It is concluded that a 1-h bout of moderate-intensity exercise performed shortly before bed elevates core body temperature before and during sleep; however, this elevated temperature does not disrupt behavioral measures of sleep obtained in the home environment in physically active male college students who were somewhat sleep deprived. PMID- 9748086 TI - Alterations in swim stress-induced analgesia and hypothermia following serotonergic or NMDA antagonists in the rostral ventromedial medulla of rats. AB - Serotonergic, NMDA, or opioid antagonists in the rostral ventromedial medulla (RVM) reduce morphine analgesia elicited from the periaqueductal gray (PAG). Continuous (CCWS) and intermittent (ICWS) cold-water swims elicit respective naltrexone-insensitive and naltrexone-sensitive analgesic responses. CCWS analgesia is reduced by systemic NMDA receptor antagonism and by systemic, but not intrathecal serotonergic antagonism. ICWS analgesia is reduced by both systemic and intrathecal serotonergic antagonism, but unaffected by systemic NMDA antagonism. The present study evaluated whether serotonergic (methysergide: 5-10 microg) or competitive [AP7 (2-amino-7-phosphonoheptanoic acid): 0.01-0.1 microg] or non-competitive [MK-801 (dizocilipine maleate): 0.3-3 microg] NMDA antagonists in the RVM altered CCWS and ICWS analgesia and hypothermia as well as basal nociceptive latencies. Methysergide in the RVM significantly potentiated CCWS, but not ICWS analgesia. In contrast, AP7 in the RVM significantly potentiated ICWS analgesia. Antagonist-induced changes in either hypothermia or basal nociception failed to account for any alterations in stress-induced analgesia. These data suggest that serotonergic, but not NMDA, receptors in the RVM may mediate collateral inhibition between mesencephalic morphine analgesia and naltrexone-insensitive CCWS analgesia. PMID- 9748087 TI - Effects of dopaminergic drugs on locomotor activity in teleost fish of the genus Oreochromis (Cichlidae): involvement of the telencephalon. AB - Single Oreochromis niloticus and O. mossambicus were placed in an unfamiliar white basin for 21 min, and their activity in this open-field situation was recorded from overhead on video. Apomorphine added to the water (2-8 mg/liter) caused a significant increase in locomotor activity, as assessed by the frequency that a fish swam over a rectilinear array of lines drawn on the base. This effect was attenuated by chlorpromazine (2 mg/liter) and abolished by the D1 antagonist SCH-23390 (1 mg/liter); the D2 antagonist metoclopramide (8 mg/liter) had no effect. Removal of both hemispheres of the telencephalon abolished the response to apomorphine, whereas removal of only one hemisphere or cauterization of the nostrils had no effect. It is concluded that the role of the dopaminergic system in the regulation of locomotor activity is reminiscent of the mammalian mesolimbic, rather than the nigrostriatal, system but that further studies are required to determine the source of the dopaminergic innervation and its likely telencephalic targets. PMID- 9748088 TI - Untrained mice discriminate MHC-determined odors. AB - Immune recognition occurs when foreign antigens are presented to T-lymphocytes by molecules encoded by the highly polymorphic genes of the major histocompatibility complex (MHC). House mice (Mus musculus) prefer to mate with individuals that have dissimilar MHC genes. Numerous studies indicate that mice recognize MHC identity through chemosensory cues; however, it is unclear whether odor is determined by classical, antigen-presenting MHC loci or closely linked genes. Previous studies have relied on training laboratory mice and rats to distinguish MHC-associated odors, but there are several reasons why training experiments may be inappropriate assays for testing if MHC genes affect odor. The aim of this study was to determine whether classical MHC genes affect individual odors and whether wild-derived mice can detect MHC-associated odors without training. In the first experiment, we found that wild-derived mice can be trained in a Y-maze to detect the odors of mice that differ genetically only in the MHC region. In the second and third experiments, we used a naturalistic habituation assay and found that wild-derived mice can, without training, distinguish the odors of mice that differ genetically only at one classical MHC locus (dm2 mutants). PMID- 9748089 TI - Time-course studies on plasma glucose, insulin, and cortisol in sea bass (Dicentrarchus labrax) held under different photoperiodic regimes. AB - Daily variations of insulin, cortisol, and glucose are studied in animals adapted to two different photoperiodic regimes, with the intervals between feeding times and photoperiodic events kept constant. Data support the existence of a daily rhythm of plasma glucose which seems to be photoperiodic. In contrast, the daily patterns of insulin in sea bass seem to be mainly influenced by feeding time; however, an effect of photoperiod can not be excluded. When the digestive tract is absent of food, insulin levels are generally minimal at feeding times and maximal during the inter-meals periods, suggesting the central control of insulin secretion during short-term food deprivation. Contrarily, the nadir values of plasma cortisol were reached at midday during the inter-meal period and peak plasma levels were evident at both light onset and offset. Disruption between metabolite and hormone patterns suggest that they are under different controls. Such results could be explained under the existence of a multioscillator system, including a food entrainable oscillator in addition to the master light entrainable oscillator. PMID- 9748091 TI - Albumin enhances sleep in the young rat. AB - Rats 4 to 7 days after weaning received intraperitoneal (i.p.) injections of vehicle (baseline day), and either serum (2 mL of lyophilized rabbit serum), 140 mg of rat albumin, or hyperosmotic NaCl (experimental day). Injections were given 1 h before light onset. Sleep-wake activity and cortical brain temperature were recorded during the subsequent 12-h light period. The intensity of non-rapid eye movement sleep (NREMS) was characterized by the power density values of the electroencephalogram slow-wave activity. The sera and albumin preparations enhanced both NREMS and slow-wave activity for 5 to 6 h starting during Hour 2 after light onset. Rapid eye movement sleep (REMS) tended to decrease. Modest (0.6 degrees C maximum deviation) biphasic changes were observed in cortical brain temperature with initial decreases for 3 h followed by rises between Hours 3 and 9 of the light period. There were no differences in the sleep responses to albumin between male and female rats. Albumin also enhanced NREMS in young rats on a protein-rich diet. A significant negative correlation was found between the NREMS promoting activity of albumin injections and the body weight of the rats. NaCl solution with the same osmolarity as that of the albumin solution failed to alter sleep. I.p. albumin injection elicited significant increases in the concentrations of cholecystokinin-like immunoreactivity in the plasma. Sleep promoting materials (hormones) in the albumin fraction, the calorigenic or nutritional value of proteins, the release of somnogenic cytokines by albumin, or endogenous humoral mechanisms stimulated by proteins (e.g., cholecystokinin or the somatotropic axis) might mediate the enhanced sleep after albumin. PMID- 9748090 TI - A search for the metabolic signal that sensitizes lateral hypothalamic self stimulation in food-restricted rats. AB - Food deprivation and restriction increase the rewarding potency of food, drugs of abuse, and electrical brain stimulation. Based on evidence that the rewarding effects of these stimuli are mediated by the same neuronal circuitry, lateral hypothalamic self-stimulation (LHSS) was used to investigate the involvement of various metabolic signals in the sensitization of reward. In Experiment 1, glucoprivation with 2-deoxy-d-glucose (150 mg/kg, intraperitoneally (i.p.)) and lipoprivation with nicotinic acid (150 mg/kg, subcutaneously (s.c.)), individually and in combination, failed to affect the LHSS threshold in ad lib. fed rats. These results suggest that signals associated with acute shortage of metabolic substrate do not sensitize reward. Because numerous responses to more prolonged negative energy balance are mediated by neuropeptide Y (NPY), the effect of exogenous neuropeptide Y upon LHSS was investigated in Experiment 2. Intraventricular infusion of orexigenic neuropeptide Y doses (2.0, 5.0, and 12.5 g), in ad lib.-fed rats, had no effect on LHSS threshold. In Experiment 3, other concomitants of prolonged negative energy balance--high circulating levels of free fatty acids (FFA) and beta-hydroxybutyrate (HDB)-were investigated. Nicotinic acid (250 mg/kg, s.c.), which suppressed serum HDB and FFA levels, had no effect on LHSS in food-restricted or ad lib.-fed rats. Mercaptoacetate (68.4 mg/kg, i.p.), which suppressed serum HDB levels and exacerbated the elevation of FFA levels, also had no effect. Thus, the brain reward system, if modulated by these substances, is not affected by transient, though marked, changes in their levels. To investigate the effect of a sustained increase in levels of FFA and HDB, a "ketogenic" diet was employed. Although this diet produced a fourfold increase in serum HDB levels, it had no effect on LHSS thresholds. Moreover, the failure of mercaptoacetate (68.4 mg/kg, i.p.) to decrease LHSS thresholds in these rats supports the conclusion that acute shortage of metabolic substrate does not sensitize reward. Other possible mechanisms of reward sensitization, including sustained decreases in circulating insulin and leptin and increases in corticosterone, are discussed. PMID- 9748092 TI - Exercise-induced slow waves in the EEG of cats. AB - The aim of the present study was to investigate the early signs of fatigue. Cats were chronically implanted with electrodes in the frontal and occipital cortical areas, and a thermocouple was inserted into the nasal orifice to record respiratory rate. After a recovery of 10 days, the animals were trained for running on treadmill. On the day before recording, a catheter was tied into one of the common carotid arteries to record arterial blood pressure. The electroencephalogram (EEG), arterial blood pressure, and respiration were recorded continuously. At the time of deceleration of running high amplitude, slow waves appeared both in the sensorimotor and occipital cortical regions. The power spectra showed a significant increase in frequencies of 1-6 Hz in the sensorimotor cortex, and of 1-10 Hz in the occipital cortex, with a great increase in the total power. During rest the pre-running, brain activity reappeared gradually. The arterial blood pressure, the heart rate, and the respiratory rate were elevated during running, but no special changes occurred at the onset of the slow waves in the EEG. The blood glucose level was somewhat higher after the first 2-min running than the pre-running level. It is concluded that the appearance of slow waves in the EEG is an early manifestation of fatigue. The cardiorespiratory changes and the blood sugar concentrations play no role in the slowing of the electrocorticogram. The present results show the involvement of brain mechanisms in the onset of tiredness. PMID- 9748093 TI - Relations between oral stereotypies, open-field behavior, and pituitary-adrenal system in growing dairy cattle. AB - The aim of this study was to investigate possible differences between calves with or without stereotypies concerning their behavioural reaction to an acute stress situation such as an open field and their behavioural and pituitary-adrenal responses to long-term tethering. Behavioural observations, open field tests, sampling for baseline adrenocorticotropic hormone (ACTH), and adrenocortical response tests after synthetic ACTH administration were made on 48 4- to 7-months old dairy calves housed in tether stalls. Behavioural observations and blood sampling for baseline ACTH and cortisol determination after synthetic ACTH were repeated a year later in the same animals. Individual stereotypy levels showed a high correlation between calf values and heifer values (p < 0.001). Baseline ACTH in the calves was related to individual stereotypy levels (p < 0.05) in that the calves with higher stereotypy levels had lower ACTH values. The release of cortisol after injection of synthetic ACTH was considerably higher in the animals as heifers than when they were calves (p < 0.001). There was a relation between adrenocortical response to ACTH and stereotypy level in the heifers, showing that the higher the stereotypy level, the lower the cortisol response (p < 0.05). In the open field tests, the calves with the highest stereotypy levels moved around least but explored most. In conclusion, this study shows that growing dairy cattle with relatively high levels of oral stereotypies differ from individuals devoid of, or with low stereotypy levels, in behavioural response patterns to a short-term stressor such as an open field in adreno-cortical responses to exogenous ACTH and in baseline ACTH after 2 weeks of tethering. PMID- 9748094 TI - Short-term effects of macronutrient preloads on appetite and energy intake in lean women. AB - This study investigated the relative satiating hierarchy of the four energy providing macronutrients (fat, carbohydrate (CHO), protein, and alcohol) in lean women. On four separate occasions, the composition of an iso-energetic lunch preload was manipulated in 12 lean (BMI < 25 kg/m2) women. The four treatments comprised a 1-MJ baseline meal and drink (40% fat, 48% CHO, 12% protein) to which was covertly added: 1) + 1MJ protein; 2) + 1MJ alcohol; 3) + 1MJ CHO; and 4) + 1MJ fat. Prior to and at 30-min intervals, subjects completed 100-mm visual analogue scales rating subjective hunger and satiety. Ninety min following the preload, an ad lib. lunch meal was given (40% fat, 48% CHO, and 12% protein) and energy intake (EI) measured. Energy intake at the lunch meal was 2195 (880, SD) kJ, 2772 (1191, SD) kJ, 2502 (681, SD), kJ and 2558 (1050, SD) kJ for the protein, alcohol, CHO, and fat preloads, respectively. There was no significant difference between the pleasantness of the preloads (p > 0.05). Macronutrient composition had a significant effect on short-term hunger (F = 3.19; p < 0.05), subjects being less hungry after the protein preload. Subjects also had a lower energy intake after the protein preload (F = 3.11; p < 0.05). We conclude that only protein has a differential short-term satiating effect when incorporated iso energetically and at a similar energy density into the diet. PMID- 9748095 TI - Temporal dynamics of corticosterone elevation in successive negative contrast. AB - A negative contrast effect in consummatory behavior that occurred when rats were shifted from 32% to 4% sucrose was accompanied by elevations in corticosterone that were measurable at 10 and 20 min but not at 40 min after the second postshift day. No corticosterone elevations were found to accompany contrast at 10 or 15 min after the first postshift day in these experiments nor immediately after the first postshift day in an earlier experiment. The present study also found an anticipatory elevation in plasma corticosterone in shifted animals just before their placement in the apparatus on the second postshift day. These results are discussed in terms of a multistage hypothesis of successive negative contrast. The anticipatory elevation is discussed in terms of uncontrollability and unpredictability of aversive events. PMID- 9748096 TI - Involvement of GABA(A) and GABA(B) receptors in the mediation of discriminative stimulus effects of gamma-hydroxybutyric acid. AB - The present study was designed to further investigate the pharmacological profile of the discriminative stimulus effects of gamma-hydroxybutyric acid (GHB). Drugs acting at the gamma-aminobutyric acid (GABA)B receptor (baclofen and CGP 35348), GABA(A)/benzodiazepine receptor complex (diazepam), N-methyl-D-aspartate (NMDA) receptor complex (dizocilpine), and cannabinoid receptor (WIN 55,212-2) were tested for substitution or blockade of the GHB interoceptive cue in rats trained to discriminate either 300 or 700 mg/kg of GHB i.g. from water in a T-maze, food reinforced drug discrimination paradigm. Baclofen completely substituted for both training doses of GHB; however, its potency in substituting for GHB increased as the training dose of GHB was increased. CGP 35348 partially and completely blocked the cue elicited by 300 and 700 mg/kg of GHB, respectively. In contrast, diazepam partially substituted for 300 mg/kg of GHB, while failing to produce a GHB-appropriate response in the rat group trained to the higher GHB dose. Neither dizocilpine nor WIN 55,212-2 substituted for GHB. Collectively, these data suggest that: a) GHB produces a compound stimulus; and b) GABA(B)- and GABA(A) mediated cues are prominent components of the mixed stimulus of GHB. However, the quality (i.e., the proportion of the component cues) of the stimulus varies as the training dose of GHB is increased; indeed, the contribution of the GABA(A)- and GABA(B)-mediated cues were smaller and greater, respectively, at 700 and 300 mg/kg of GHB training doses. PMID- 9748097 TI - Effects of strawbedding on physiological responses to stressors and behavior in growing pigs. AB - To study the effects of environmental enrichment on physiological responses to stressors and behavior in growing pigs, pigs were housed in either a poor environment (standard farrowing pens followed by standard rearing and fattening pens) or in an enriched environment (larger farrowing pens followed by larger rearing and fattening pens, provision of straw). Body temperature, heart rate and salivary cortisol were measured during baseline conditions and in response to relocation, isolation and restraint. Pigs housed in the poor environment performed more manipulative social behavior directed to penmates than pigs housed in the enriched environment. Physiological responses to the stressors were the same for enriched- and poor-housed pigs. Surprisingly, enriched-housed pigs had significantly higher baseline salivary cortisol concentrations, especially at 14 and 17 weeks of age. Moreover, enriched housed pigs had a lower baseline body temperature at 17 weeks of age. Thus, provision of straw has an effect on behavior, baseline HPA-axis activity and baseline body temperature in growing pigs. PMID- 9748098 TI - ACTH and beta-endorphin in transcendental meditation. AB - We have evaluated the effect of Transcendental Meditation (TM) on the hypothalamo hypophyseal-adrenal axis diurnal rhythms through the determination of hormone levels. Blood samples were taken at 0900 hours. and at 2000 hours. These samples were taken from 18 healthy volunteers who regularly practice TM and from nine healthy non-meditators. Cortisol, beta-endorphin, and adrenocorticotropic hormone (ACTH) were measured at both hours. TM practitioners showed no diurnal rhythm for ACTH and for beta-endorphin (ACTH, pg/mL, mean +/- SE; 13.8+/-1.2 - 12.1+/ 1.5/beta-endorphin, pg/mL; 14.4+/-1.5 - 17.2+/-1.9, at 0900 hours and 2000 hours, respectively), in contrast to control subjects, who showed normal diurnal rhythm for these hormones and for cortisol (ACTH, pg/mL; 19.4+/-1.9 - 11.9+/-2.2/beta endorphin, pg/mL; 25.4+/-1.7 - 17.7+/-1.1/Cortisol, ng/mL; 201.4+/-13.2 - 71.3+/ 6.5, at 0900-2000 hours, respectively, p < 0.01 in the three cases). Practitioners of TM with similar anxiety levels to those of the control group showed a different pattern in the daytime secretion of pituitary hormones. TM thus appears to have a significant effect on the neuroendocrine axis. Because cortisol levels had a normal pattern in the TM group, these results may be due to a change in feedback sensitivity caused by this mental technique. PMID- 9748099 TI - Influence of the composition of a meal taken after physical exercise on mood, vigilance, performance. AB - The metabolic and behavioral effects of nutrients after exercise on vigilance level, performance, and mood have been minimally studied and have given contradictory results. In order to increase the understanding of the relationships between nutrition, exercise and performance, this experiment compared the effects on mood and performance of a protein- rich meal and a protein- poor meal, eaten just after an acute session of exercise. Vigilance and mood were evaluated by visual analog scales, and memory was measured by memory search task from the AGARD STRES battery, based on the Sternberg paradigm. Forty two subjects were involved in this experiment. All subjects participated in the study of the effect of exercise after two kinds of meals (protein and nonprotein). Two groups of fourteen subjects we used to evaluate the effect of the exercise and the effect of the delay of meal intake after exercise in the two kinds of diet. The results show no difference in memory performance between exercise and rest conditions, nor between "protein" and "no protein" meal groups. They do show, however, that subjects feel happier after a meal with protein than after a meal without protein. The effects of the "no protein" meal on drowsiness differ with the glucide content of the meal. Subjects are less drowsy when they eat between 125 and 150 g of glucide than when they eat more than 150 g. The rousing effect induced by physical exercise is counterbalanced when subjects eat more than 150 g of carbohydrate. The anxiolytic effect of glucide is re established. PMID- 9748100 TI - Consumption of electrolytes and quinine by mouse strains with different blood pressures. AB - Daily fluid intakes were measured using two-bottle tests in female mice of inbred strains with high (BPH/2), normal (BPN/3) or low (BPL/1) blood pressure. The mice were offered a choice between water and different concentrations of NaCl (37.5 600 mM), KCl (1-400 mM), CaCl2 (1-100 mM) and quinine hydrochloride (0.003-1.0 mM). Compared with the normotensive strain, the hypertensive mice had higher water and total fluid intakes, and lower intakes of NaCl, KCl (only 200 mM) and quinine; the hypotensive mice had higher intakes of KCl (only 10-50 mM) and lower intakes of CaCl2 and quinine. These data suggest that fluid and salt intake are not linearly related to blood pressure, but are independently determined in these strains. Certain concentrations of the salts were preferred relative to water, which depended on mouse genotype: the BPN/3 and BPL/1 mice strongly preferred 37.5-150 mM NaCl, the BPL/1 mice preferred 10-100 mM KCl, and the BPN/3 mice preferred 1-10 mM CaCl2. PMID- 9748101 TI - Neural substrates for leptin and neuropeptide Y (NPY) interaction: hypothalamic sites associated with inhibition of NPY-induced food intake. AB - Intracerebroventricular (i.c.v.) injection of leptin, the adipocyte hormone, inhibits neuropeptide Y (NPY)-induced feeding in the rat. To identify the neural substrate for leptin and NPY interaction in the hypothalamus, we evaluated the expression of c-fos-like immunoreactivity (FLI), an early marker of neuronal activation, in response to icv administration of leptin, NPY and leptin plus NPY. As expected, leptin significantly decreased NPY-induced feeding in leptin plus NPY-treated rats. A comparative evaluation of the number of FLI-positive neurons in hypothalamic sites showed that both leptin and NPY activated FLI in the parvocellular subdivision of the paraventricular nucleus (pPVN), dorsomedial nucleus (DMN) and ventromedial nucleus (VMN). NPY also augmented the FLI response in the magnocellular PVN (mPVN) and supraoptic nucleus (SON), two sites where leptin alone was ineffective. Combined leptin and NPY treatment significantly decreased the number of FLI-positive neurons in the magnocellular PVN but increased their number in the dorsomedial nucleus as compared to the number of FLI-expressing neurons in these sites after NPY and leptin alone. Because there is morphologic evidence of a link between magnocellular PVN and dorsomedial nucleus, these results suggest the functional involvement of leptin plus NPY responsive elements in these sites in reduction of NPY-induced feeding by leptin. PMID- 9748102 TI - Effects of angiotensin II on sexual function, blood pressure, and fluid intake are differentially affected by AT-1 receptor blockade. AB - We have previously reported that third ventricular administration of angiotensin II (ANG II) immediately before mating tests suppressed copulatory behavior in male rats. The present studies examine the effects of short- (3 days) and long term (21 days) intracerebroventricular (i.c.v.) infusion of ANG II (6 microg/h), on parameters of copulatory behavior, fluid intake, and blood pressure in sexually experienced male Long Evans rats. Further, to test the hypothesis that suppression of masculine copulatory behavior by ANG II involves interaction with the angiotensin AT-1 receptor, a highly selective nonpeptide antagonist (L 158,809) was administered in the drinking water (25 mg/liter) to a group of ANG II-infused animals. I.c.v. infusion of ANG II was associated with increases in systolic blood pressure and fluid intake. In copulatory tests after 3, 9 and 15 days of infusion, rats infused with ANG II exhibited increased latencies to the initiation of copulatory behavior and to ejaculation, as well as increased intervals to reinitiate copulatory behavior after the ejaculation. Administration of L-158,809 blocked the effects of i.c.v. infusion of ANG II on systolic blood pressure and fluid intake. Further, L-158,809 attenuated the effects of i.c.v. infusion of ANG II on parameters of copulatory behavior. Data from this study provide support for a modulatory role for ANG II in the regulation of sexual behavior. In addition, this regulation seems to involve the AT-1 receptor. PMID- 9748103 TI - Effects of age and ethanol on dopamine and serotonin release in the rat nucleus accumbens. AB - Neural functions in the nucleus accumbens (ACC) play an important role in alcohol drinking behavior. In the present study, we observed the effects of age and ethanol (EtOH) on dopamine (DA) and serotonin (5-HT) release in the ACC of freely moving 4-, 10-, and 16-month-old rats using brain microdialysis techniques. After co-perfusion with 200 mM ethanol, ACC DA, and 5-HT release were decreased significantly in 16-month-old rats compared to those at 4 months old. ACC DA and 5-HT neurons of aged rats were less sensitive to ethanol. On the other hand, both basal extracellular DA and 5-HT release in the ACC were significantly higher in 16-month-old than in 4-month-old rats. Therefore, aging results in opposite changes in basal and alcohol-induced DA and 5-HT release in the ACC. PMID- 9748104 TI - Effects of social status after mixing on immune, metabolic, and endocrine responses in pigs. AB - The effects of social rank on immune, metabolic, and endocrine responses were studied in 10 newly mixed groups of German Landrace pigs (9 individuals each) at an age of 12 weeks. Immediately after mixing, the agonistic interactions (AI) of all group members were continuously recorded over 3 days (10 h daily). An individual dominance value (DV) was calculated by the number of wins minus defeats in relation to all decisive fights (DV < or = 0, subordinate; DV > 0, dominant). Blood samples were taken 24 h before and 3 days after mixing. The data showed that the social status had a significant effect on lymphocyte proliferation in responses to different mitogens: socially dominant pigs had higher proliferative response than subordinate pigs. In addition, during the observation period the lymphocyte activation by mitogens increased in the dominant animals and decreased in the subordinate animals with increasing number of agonistic interactions. The rise in total serum IgG concentration 3 days after mixing was higher in dominant pigs compared with subordinates. The dominance status did not significantly affect plasma metabolic levels nor cortisol concentrations. However, mixing appeared to increase glucose and total protein values and to decrease alkaline phosphatase and cortisol levels in both, dominant and subordinate pigs. In conclusion, mitogen induced cell proliferation seems to be a valuable marker for acute social stress in pigs. PMID- 9748105 TI - Subdiaphragmatic vagotomy does not block sleep deprivation-induced sleep in rats. AB - Cytokines, such as interleukin-1beta (IL-1beta), are involved in physiological sleep regulation and in the sleep responses to sleep deprivation. Sleep deprivation increases systemic cytokine levels and recent evidence suggests that cytokine-to-brain communication occurs via the vagus nerve. Furthermore, the vagus nerve plays a role in sleep responses elicited by feeding and vagal activity affects electroencephalographic (EEG) activity. Thus, this study examined sleep-wake activity and brain temperature (Tbr) responses to sleep deprivation in subdiaphragmatically vagotomized and sham-operated rats. In control rats, 6 h of total sleep deprivation significantly increased nonrapid eye movement sleep (NREMS), rapid eye movement sleep (REMS), and electroencephalographic slow-wave activity during nonrapid eye movement sleep. Brain temperature was significantly increased during the 6 h of sleep deprivation and decreased following sleep deprivation. Vagotomy had no significant effects on any of these variables. These results indicate that the subdiaphragmatic vagus nerve is not critical in the sleep and thermoregulatory responses after 6 h of sleep deprivation. Together with other data, the current results suggest that central pools of interleukin-1 are important in moderate sleep deprivation induced sleep responses and that vagotomy does not disrupt the ability to increase sleep using a well-known sleep-inducing stimulus likely mediated by brain cytokines. PMID- 9748106 TI - Intracerebroventricular glucagon-like peptide-1 (7-36) amide inhibits sham feeding in rats without eliciting satiety. AB - Glucagonlike peptide-1 (7-36) amide (GLP-1) and its receptors are present in several brain regions and may play a role in the physiological control of feeding. To investigate the effect of GLP-1 on eating in the absence of postingestive food stimuli, rats were implanted with gastric cannulas for sham feeding and lateral ventricular cannulas for infusion of GLP-1. Rats (n = 10) sham fed 0.8 mol/L sucrose for 45 min, beginning 5 min after intracerebroventricular (icv) infusion of 2.5 microL of artificial cerebrospinal fluid with 0-30 microg of GLP-1 . Behaviors were observed each minute using a time-sampling technique. Additionally, lick-by-lick records of the microstructural pattern of sucrose intake were made during the first 15 min of each test for five rats receiving 3 and 10 microg of GLP-1. GLP-1 decreased sham fed intake by as much as 50%, but GLP-1 did not terminate sham feeding. The frequency of observations of feeding was decreased, but the frequency of resting, the terminal item in the behavioral sequence of postprandial satiety in real feeding rats, did not reliably increase. No abnormal behaviors were observed. Although GLP-I did not affect the latency to begin sham feeding, it significantly reduced the initial rate of licking. GLP-I did not affect the motor aspects of licking, because the interlick intervals within individual bursts of licking or overall lick efficiency were normal. These data suggest that intracerebroventricular infusions of GLP-1 inhibit sham feeding by decreasing the orosensory positive feedback that drives licking, rather than by activating physiological satiating mechanisms or nonspecific mechanisms such as aversion or motor incapacity. PMID- 9748107 TI - Medial prefrontal lesion deficits involving or sparing the prelimbic area in the rat. AB - The rat medial prefrontal cortex (PFC) is believed to play a central role in working memory and selective attention processes. More recently, it has been shown that the effects of large PFC lesions on working memory may be due to the prelimbic area of the PFC. The aim of the present study was to compare the effects of lesions of the prelimbic area with PFC lesions that involved or spared the prelimbic area on shuttlebox avoidance and radial maze learning in rats. The findings indicate that rats with PFC lesions that spared the prelimbic area were impaired at avoidance but not radial arm maze learning, whereas rats with prelimbic lesions or PFC lesions that included this area were impaired on the radial arm maze but not the avoidance learning task. Results support the notion that the medial frontal cortex of the rat is a functionally dissociable region and suggest that the prelimbic area appears to be critical for working memory, but less so for attention processes. PMID- 9748108 TI - Ketamine blocks a conditioned taste aversion (CTA) in neonatal rats. AB - These experiments explored the effects of glutamate, N-methyl-D-aspartate (NMDA) receptor blockade on the formation, retention, and expression of conditioned taste aversion (CTA) in young rats. Previous data from our laboratory suggested that ketamine administration potentiates a CTA in E18 rat fetuses. The current studies investigated this phenomenon in neonates. High-pressure liquid chromatography (HPLC) methods were used to determine the amount of ketamine that must be injected intraperitoneally (i.p.) to achieve brain ketamine levels in neonates comparable to those found in the fetuses from our previous experiments. Then, on their day of birth, Sprague-Dawley rat pups received injections of either 0.1, 10, or 70 mg/kg of ketamine HCI, i.p. or a Sal control injection. One half hour later, pups were injected orally with either Saccharin (Sac; 10 microL of 0.3%) or water followed by an injection of either lithium chloride (LiCl; 81 mg/kg) or Sal (i.p.). The CTA was evaluated in two different tests. Two weeks after conditioning, the dam was anesthetized and the frequency with which pups attached to Sac-painted nipples versus nipples painted with water was measured (i.e., the nipple taste test, NTT). Controls for state-dependent learning were run in which 10 mg/kg of ketamine or saline (Sal) was administered before both taste aversion conditioning and the NTT. After weaning, the CTA was also evaluated by measuring the amount of Sac (0.3%) or water consumed during a two bottle test. Neonates that received Sal control injections before the Sac + LiCl pairing acquired CTAs and avoided Sac-painted nipples. However, the pups injected with ketamine on the conditioning day only (P0) did not avoid Sac-painted nipples (as compared to controls). Pups that had ketamine both at the time of CTA training and testing, or just before the NTT, also failed to avoid Sac-painted nipples. Ketamine's acute effects apparently influenced the outcome of the NTT of state-dependent control subjects. Rat pups that received the highest doses of ketamine (10 or 70 mg/kg) and tasted Sac on P0 later failed to show a neophobia for Sac-painted nipples. Whereas, rat pups that received the high dose of ketamine and water on P0, later exhibited a neophobic response. These data suggest that ketamine did not impair the animal's ability to taste Sac. These data reflecting a ketamine-induced blockade of neonatal CTAs may be contrasted with our previous findings in which ketamine potentiated fetal CTAs. However, they are in consonance with data from adult rats suggesting that ketamine can cause an amnesia for CTAs. NMDA receptor blockade may shape memory formation in a manner that is dependent on the stage of brain development. PMID- 9748109 TI - Manipulation of NMDA-receptor activity alters extinction of an instrumental response in rats. AB - The effects of pharmacologically manipulating N-methyl-D-aspartate(NMDA)-receptor activity were examined during extinction of an appetitive instrumental response in rats. After reaching acquisition criterion, subjects were treated with the antagonist dizocilpine maleate (MK801; 0.1 mg/kg), the agonist D-cycloserine (3 mg/kg), or vehicle-alone (control) and tested during a non-reinforced (extinction) session. The antagonist decreased the average number of responses occurring during the test session whereas the agonist increased the average number in contrast to controls. The effect on retention performance may be mediated by differential influence on the N-methyl-D-aspartate-dependent synaptic plasticity that occurs during associative learning. In conjunction with other studies, these data suggest that N-methyl-D-aspartate agonism may be an effective intervention for memory dysfunction. PMID- 9748111 TI - Plasma cocaine levels and locomotor activity after systemic injection in virgin and in lactating maternal female rats. AB - We have determined the temporal pattern of plasma cocaine levels and increased activity that result from acute systemic injections of cocaine to female rats in two different endocrine and behavioral states, in nonmaternal virgins and in lactating maternal dams. Plasma levels of cocaine as well as ambulatory and rearing activity were determined every 30 min for a total of 300 min after subcutaneous injections of either 10, 20, or 40 mg/kg of cocaine. Virgin females had no prior drug history, whereas lactating, maternal dams had received two cocaine injections before activity testing. Within 30 min after an injection, cocaine in the plasma and activity were substantially elevated, and generally remained so for 270-300 min. Overall, plasma cocaine levels and activity were well correlated and followed a predictable dose-response pattern. The onset, peak, duration, and decline of activity corresponded generally to the onset, peak, duration, and decline of plasma cocaine. For virgins, mean ambulatory activity increased 2.5-4.0-fold over baseline, whereas in lactating females activity increased 5-11-fold over baseline. Stereotypy did not occur. Although the general responsivity of these females to cocaine was very similar to that reported for males, there are differences in the timing of peak activity and the return of activity to baseline when the virgins and the lactating dams are compared to each other and to reports by others on male rats. These data support the hypothesis that endocrine or behavioral state may influence the responsiveness of animals to cocaine. PMID- 9748110 TI - Conditioned inhibition of antibody response and CD4 positive cells. AB - Animals were assigned to three experimental groups, conditioned (cyclophosphamide glucose treatment; C group), non-conditioned (cyclophosphamide-commercial pellet treatment; NC group) and placebo (saline solution-glucose treatment; P group). The three groups were injected intraperitoneally (i.p.) with sheep red blood cells. An hemagglutination assay according to standard procedures was performed along with the flow cytofluorometric analysis of leukocyte surface antigens CD4, CD8 and CD45. C group consumed less food on Days 3 and 6 than on Day 0, NC animals showed no changes in food consumption throughout the experimental period, whereas an increasing trend was observed for P animals. The proportion of T lymphocytes expressing CD8 and CD45 did not differ significantly among C, NC and P groups. Group C showed the lowest proportion of T lymphocytes bearing CD4, whereas Group P displayed the highest. The antibody response was lower in the Group C than in the groups NC and P. We conclude that, although the mechanisms by which humoral conditioned immunosuppression occurs in mice is still unknown, a reduced T helper-mediated activation of B-cells may play an important role in producing conditioned humoral response. PMID- 9748112 TI - The role of specific macronutrient availability in the effect of food restriction on length of lactational diestrus in rats. AB - In lactating rats, food restriction for the first two weeks postpartum extends the period of lactational diestrus by about 1 week. In these studies we investigated whether this effect results from caloric restriction or the reduced availability of a specific macronutrient. In Experiment 1 lactating rats nursing litters of eight pups were assigned to one of four conditions: 1) ad lib. fed; 2) protein-restricted; 3) carbohydrate-restricted; and 4) fat-restricted. Animals in all the restricted conditions were given access to 50% of ad lib. intake of the appropriate nutrient for Days 1-14 postpartum and ad lib. access to the other two macronutrients. In Experiment 2, ad lib. supplementation from one macronutrient source was provided to lactating rats given restricted access to a composite diet. No differential effect of specific macronutrient deprivation or supplementation on length of lactational diestrus was observed in these studies. Thus, the results of both studies are consistent with the hypothesis that caloric restriction plays a primary role in inducing the prolongation of lactational diestrus in food-restricted rats. PMID- 9748113 TI - Increased GABA-gated chloride ion influx in the hypothalamus of low-anxiety rats. AB - While it is generally accepted that y-aminobutyric acid type A (GABA(A))-receptor agonists decrease anxiety by facilitating the neuronal influx of chloride, the site of action within the brain is not clearly delineated. To gain further insight into the locus of anxiolytic action of GABA in the brain, we measured the distribution of GABA-stimulated chloride influx in seven regions of the brain from high- and low-anxiety rats. Our results show a significant increase in GABA gated chloride influx in the hypothalamus of rats exhibiting low anxiety. The role of the hypothalamus in the regulation of anxiety is briefly discussed. PMID- 9748114 TI - New apparatus for studying behavioral thermoregulation in rats. AB - The operant system described here contains a box that can be convectively heated or cooled. A rat moves freely in the box. Its location is monitored photoelectrically while its deep body temperature is monitored by a telemetry system. In heat-escape experiments, hot air (40 degrees C) flows through the box. When the rat enters a reward zone the air source is switched and cold air (0 degrees C) flows through the box for a given period (30 s). Conversely, in cold escape experiments cold air flows through the box and when the rat enters the reward zone the air source is switched to a warm one. Experiments show that rats quickly learn to stay near the reward zone and move in and out of it periodically. This system is based on behavior more natural than the frequently used lever-pressing response, and has many advantages for use in studies involving behavioral thermoregulation. PMID- 9748115 TI - Mammalian bombesin-like peptides extend the intermeal interval in freely feeding rats. AB - We evaluated the effects of systemic delivery of amphibian bombesin and its mammalian homologues on the length of the postprandial intermeal interval. Adult male rats, feeding ad libitum, were injected intraperitoneally (i.p.) 5 min after the end of the first nocturnal meal with 0 (vehicle), 2.5, 5, or 10 nM/kg of tetradecapeptide bombesin (BN), gastrin-releasing peptide(1-27) (GRP1-27), the C terminal decapeptide of GRP(18-27) (GRP18-27), the C-terminal decapeptide of neuromedin B (NMB23-32), or combinations of equimolar doses of GRP1-27 and NMB23 32. BN produced a potent, dose-related extension (maximum of 177%) of the first postprandial intermeal interval; GRP1-27 produced a lesser but significant prolongation (maximum of 47%); the combination of GRP1-27 and NMB23-32 produced an intermediate prolongation (maximum of 70%); GRP18-27 alone and NMB23-32 alone failed to produce any significant change. Peptide effects were limited to the first postprandial intermeal interval. The results demonstrate that systemic, postprandial injection of BN, GRP1-27, or the combination of GRP1-27 and NMB23-32 extends the duration of the postprandial intermeal interval. The results suggest that the endogenous peptides, released in the gastrointestinal tract by ingested food, have a potent satiety action, selectively lengthening the intermeal interval. PMID- 9748116 TI - The relationship between high-dose treatment and combination chemotherapy: the concept of summation dose intensity. AB - The most important variables for the clinical use of antitumor agents (AAs) are dose and combination chemotherapy. The objectives of this study were to analyze the relationship between these two variables and to propose a unified conceptual framework for the construct and interpretation of clinical trials. Definitions and variables with respect to dose include potency, therapeutic index, standard dose, efficacy, relative efficacy, dose-limiting toxicity (DLT), dose rate, dose density, dose intensity, and fractional dose intensity. Our overarching concept, that is, summation dose intensity (SDI), was calculated in several ways, depending upon the nature of the data, and included the relative efficacy method, the unit regimen method, and the high dose method. The SDI concept was then applied to disease categories and strategies to determine its usefulness and effectiveness in integrating dose and combinations. The tumors and settings were: mustargen-vincristine-procarbazine-prednisone in Hodgkin's disease, combination chemotherapy for acute lymphocytic leukemia in children, metastatic breast cancer including dose and combinations, selected other solid tumors, alternating chemotherapy, and high dose studies in the leukemias and lymphomas. SDI was effective in integrating and quantifying dose and combination chemotherapy. For classical AAs, the implication of SDI for the construct and analysis of clinical trials was emphasized. In addition to new drug development, emphasis should be given to reducing or eliminating DLTs, such as those of the marrow, now and, in the future, those of the gastrointestinal tract toxicity and other DLTs. The above was derived from and applies to the classical AAs. Whether they will apply to, with appropriate adjustment, agents with significantly different dose response curves, such as biotherapeutics and hormonal agents, remains to be determined. PMID- 9748117 TI - Clinical impact of pharmacokinetically-guided dose adaptation of 5-fluorouracil: results from a multicentric randomized trial in patients with locally advanced head and neck carcinomas. AB - A significant link between 5-fluorouracil (5FU) plasma concentration and its therapeutic activity has been demonstrated in colon and head and neck cancer patients for 5FU used as a continuous infusion. Dose adjustment based on pharmacokinetic follow-up has been proposed to decrease hematological and digestive toxicities, but the clinical impact of this approach has not yet been demonstrated. A randomized multicentric study was conducted to evaluate the clinical interest of 5FU dose adaptation guided by pharmacokinetics. One hundred twenty-two head and neck cancer patients were randomly assigned to receive induction chemotherapy with cisplatin (100 mg/m2, day 1) and 5FU (96-h continuous infusion), either at standard dose (St-arm; 4 g/m2) or at a dose adjusted according to the 5FU area under the curve (AUC0-48h; PK-arm). In total, 106 patients were evaluable for toxicity and response. In the PK-arm (n = 49), 5FU doses and area under the curve were significantly reduced during cycle 2 and cycle 3 (P < 0.001) as compared with the St-arm (n = 57). Grade 3-4 neutropenia and thrombopenia were significantly more frequent in the St-arm as compared with the PK-arm (17.5% versus 7.6%, respectively; P = 0.013). No grade 3-4 mucositis occurred in the PK-arm, whereas 5.1% was observed in the St-arm (P < 0.01). The objective response rate was comparable in the two treatment arms: 77.2% in the St arm versus 81.7% in the PK-arm. The present study is the first to demonstrate, in a randomized design, the clinical interest of an individual 5FU dose adaptation based on pharmacokinetic survey, in terms of therapeutic index improvement. PMID- 9748118 TI - Immunocytochemical detection of somatostatin receptors sst1, sst2A, sst2B, and sst3 in paraffin-embedded breast cancer tissue using subtype-specific antibodies. AB - The long-acting somatostatin analogue octreotide (SMS 201-995) inhibits growth of certain breast cancer cell lines in vivo and in vitro. Because the antiproliferative action of octreotide depends on at least the presence of somatostatin receptors, it is crucial to determine the pattern of somatostatin receptor protein expression on the tumor cells. In the present study, we have raised polyclonal antibodies to somatostatin receptor subtypes (ssts) sst1, sst2A, sst2B, and sst3 using peptides corresponding to their COOH-terminal sequences. These antisera were used for immunocytochemical staining of paraffin sections of 33 primary breast cancers. Somatostatin receptor-like immunoreactivity (Li) was predominantly localized to the plasma membrane of the tumor cells. In the vast majority of positively stained tumors, somatostatin receptor-Li was uniformly present on nearly all tumor cells. Both the level and the pattern of expression of ssts varied greatly between individual carcinomas. sst2A-Li and/or sst2B-Li was detectable in 28 tumors (85%); among these, 14 tumors (42%) showed particularly high levels of sst2-Li. sst1-Li was found in 17 (52%) cases and sst3-Li in 16 (48%) cases. The expression of ssts was independent of patient age, menopausal status, diagnosis, histological grade, and levels of estrogen and progesterone receptors. The immunocytochemical determination of somatostatin receptor status allows direct detection of receptor protein on the tumor cells and, hence, may provide more precise information than reverse transcription-PCR for predicting response to octreotide therapy in breast cancer. PMID- 9748119 TI - Prevention of intestinal toxic effects and intensification of irinotecan's therapeutic efficacy against murine colon cancer liver metastases by oral administration of the lipopeptide JBT 3002. AB - The induction of severe diarrhea limits the usefulness of the DNA topoisomerase I inhibitor irinotecan (CPT-11) in the treatment of advanced colon cancer. We investigated whether oral administration of the new synthetic bacterial lipopeptide, JBT 3002, encapsulated in phospholipid liposomes could prevent damage to the intestinal epithelium and lamina propria and thus allow for the parenteral administration of high-dose irinotecan to mice with established syngeneic CT-26 colon cancer liver metastases. Treatment of mice with four daily i.p. injections of 100 mg/kg irinotecan was effective against liver metastases but also resulted in loss of body weight and early death. Histopathological examination of the intestines after this treatment revealed loss of villi, epithelial vacuolation, decrease in the number of cells in the crypts in S-phase, increase in the number of apoptotic cells, and reduction in the number of lymphocytes in the lamina propria. In contrast, treatment of mice with the same irinotecan regimen after oral administration of JBT 3002 produced highly significant inhibition of liver metastases without detectable damage to the intestines. Studies that used irinotecan administered once a week for 3 weeks after pretreatment with oral JBT 3002 demonstrated significantly intensified eradication of established CT-26 liver metastases compared with treatment with once-weekly irinotecan alone. Histological studies revealed that the liver metastases in mice treated with oral JBT 3002 and i.p. irinotecan contained a higher number of macrophages than metastases in mice treated with either drug alone. In vitro studies revealed that irinotecan produced direct antiproliferative effects but JBT 3002 did not. Tumor cells exposed to both irinotecan and macrophages activated by JBT 3002 were highly susceptible to lysis. These data show that oral administration of JBT 3002 can prevent irinotecan-induced gastrointestinal toxic effects and maintain the integrity of the lamina propria, thus allowing for intensification of irinotecan therapy against liver metastases from colon cancer. PMID- 9748120 TI - Immunohistochemical detection of NAD(P)H:quinone oxidoreductase in human lung and lung tumors. AB - NAD(P)H:quinone oxidoreductase (NQO1) is a flavoenzyme that catalyzes the two electron reduction of quinones and related compounds. With the use of biochemical assays, NQO1 has been shown to be overexpressed in many types of cancer, including non-small cell lung cancer (NSCLC). NQO1 can bioactivate antitumor quinones such as mitomycin C, and new quinone-based drugs are currently being developed to target this enzyme in tumors such as NSCLC. Because there is no information on the cell-specific expression of NQO1 in lung, the purpose of this study was to examine the expression of NQO1 in human NSCLC, small cell lung cancer, carcinoid lung tumors, and normal lung using immunohistochemistry. A high level of NQO1 protein expression was detected by immunohistochemistry in NSCLC (adenocarcinoma, squamous cell carcinoma, and bronchoalveolar carcinoma), but no NQO1 protein could be detected in small cell lung cancer or carcinoid lung tumors. In addition, NQO1 protein expression was examined by immunohistochemistry in normal lung tissue. A high level of NQO1 protein expression was detected by immunohistochemistry in normal lung respiratory epithelium, with the highest levels of expression observed in ciliated columnar epithelial cells. Significant amounts of NQO1 protein were also detected in the vascular endothelium and adipocytes. These data demonstrate that NQO1 is overexpressed in NSCLC. Cells in normal lung also contain marked NQO1 protein and may be damaged by drugs activated by NQO1. These data validate NSCLC as a target for NQO1-directed agents and suggest that the potential for lung toxicity be considered in the preclinical development of quinone-based antitumor drugs. PMID- 9748121 TI - The effects of local hyperthermia on the catabolism of a radioiodinated chimeric monoclonal antibody. AB - Local hyperthermia has been shown to increase the tumor uptake and tumor:normal tissue ratios of radiolabeled monoclonal antibodies (mAbs) in athymic mouse xenograft models. The current study was undertaken to determine whether this behavior was related in part to alterations in mAb catabolism by local hyperthermia. Human/mouse chimeric 81C6 mAb reactive with tenascin and a nonspecific control mAb were labeled with 125I using Iodo-Gen and given to separate groups of athymic mice bearing s.c. D-54 MG human glioma xenografts. Half of the animals were then subjected to 4-h tumor-localized hyperthermia at 41.8 degrees C, a protocol previously shown to enhance the specific tumor uptake of the mAb in this xenograft model. The tumor, serum, liver, kidney, and urine were collected from heated as well as control animals 4 and 24 h after injection of the mAb and analyzed by SDS-PAGE and trichloroacetic acid precipitation. At 24 h, a significantly higher percentage of 81C6 was present as intact mAb in the tumors harvested from heated animals compared with those from controls. Unexpectedly, intact mAb was found in the urine of mice immediately after hyperthermia, but not in unheated control animals. We conclude that local hyperthermia decreases the catabolism of the mAb in the tumor and increases the urinary excretion of the mAb through a transient increase in glomerular permeability. PMID- 9748122 TI - Differential uptake of estramustine phosphate metabolites and its correlation with the levels of estramustine binding protein in prostate tumor tissue. AB - Estracyt (EMP) has been used for the treatment of hormone refractory prostate cancer for many years. Recently, new data from combination studies have given rise to new interest in this old drug. Explanations for the synergy found in the clinic are many, but one major factor may be the previous indication that the drug accumulates in the prostate tumor. We have, therefore, examined the level of the four metabolites, estromustine (EoM), estramustine (EaM), estrone, and estradiol in the tumor and serum of 14 patients with T2 and T3 prostate cancer receiving a single i.v. dose of 600 mg of EMP, about 12 h before radical prostatectomy. Because it has been suggested that the uptake into the prostate tumor is due to binding to the estramustine binding protein (EMBP), we have in addition measured the level of EMBP in the prostate tumor tissue. The main serum and tissue metabolite in all patients was EoM followed by EaM, estrone, and estradiol. The levels for EoM ranged from 63.8-162.8 ng/ml in the serum and from 64.8-1209 ng/ml in the prostate tumor, resulting in a mean ratio for serum to tumor of 1:5. The levels for EaM ranged from 8.3-51.4 ng/ml in the serum and 73.9 563.4 ng/ml in the tumor, giving a mean ratio for serum to tumor of 1:13. The levels of EMBP were higher in T3 tumors than in T2 tumors, 54.1 and 40.7 ng/g tissue, respectively. A significant correlation was found between the levels of EaM (r = 0.60) and the levels of EMBP in the tumor. These data demonstrate that 12 h after a single i.v. dose of 600 mg of EMP the levels of the cytotoxic metabolites EoM and EaM are substantially higher in the tumor than in the serum of the same patient and that a correlation exists between the levels of EaM in the tumor and the levels of EMBP. Thus, this supports the hypothesis that the EMBP is responsible for the retention of EoM and EaM in the prostate tumor. PMID- 9748123 TI - Comparison of 5-fluorouracil pharmacokinetics in patients receiving continuous 5 fluorouracil infusion and oral uracil plus N1-(2'-tetrahydrofuryl)-5 fluorouracil. AB - Plasma 5-fluorouracil (5-FU) levels were compared in the same patients after approximately equimolar doses (1.9 mmol/ m2/day) of 5-day continuous i.v. infusion of 5-FU (CIFU) and oral administration of a formulation of two combined pharmacological agents, uracil (U) plus N1-(2'-tetrahydrofuryl)-5-fluorouracil (ftorafur or FT), a prodrug of 5-FU. Ten patients received CIFU for 5 days, then, after a week wash-out period, began the 28-day oral UFT regimen, which was given in three daily divided doses. Following 1 h of CIFU, the plasma 5-FU levels reached a steady state of 0.6+/-0.2 microM (mean+/-SD; day 1), which was maintained for the entire 5-day infusion period (0.6+/-0.1 microM). In contrast, the maximum 5-FU concentrations (Cpmax) generated from oral UFT at 1 h after dose administration on days 1 and 5 were 2.1+/-1.5 microM and 2.3+/-1.9 microM, respectively, which were higher than the steady-state levels during CIFU. These high 5-FU levels disappeared with an apparent elimination half-life (tl/2,beta) of 5.2+/-2.4 h (day 1) and 7.2+/-3.9 h (day 5). On day 1 of both regimens, CIFU patients had significantly larger 5-FU area under the concentration versus time curve (AUC0-8h) values (4.4+/-1.3 microM.h) than the AUC value when they received the UFT regimen (2.6+/-1.7 microM.h; P = 0.02). However, by day 5, there were no significant differences between AUC0-8 h values in patients receiving CIFU and UFT, respectively (4.8+/-1.5 microM.h versus 3.8+/-2.2 microM.h; P = 0.30)]. On day 5, the average concentration of the metabolite 5-FU at steady-state (Css,aver) within dose interval of 8 h (0.48+/-0.28 microM) for the oral UFT treatment is comparable with the Cpss values of 5-FU from CIFU-treated patients. The post-UFT generated 5-FU pharmacokinetic parameters (higher Cp(mx, comparable Css,aver, equal AUC values, and longer apparent t1/2,beta of 5-FU) may make oral UFT a preferred method of delivering this fluoropyrimidine over CIFU. In addition, oral UFT would eliminate the incidence of venous thrombosis and catheter-related infections sometimes seen in patients treated with CIFU. Furthermore, the convenience and decreased cost of oral administration may be preferable for many patients, particularly those receiving 5-FU for palliation. PMID- 9748124 TI - In vivo inhibition of aromatization by exemestane, a novel irreversible aromatase inhibitor, in postmenopausal breast cancer patients. AB - The effect of exemestane (6-methylenandrosta-1,4-diene-3,17-dione) 25 mg p.o. once daily on in vivo aromatization was studied in 10 postmenopausal women with advanced breast cancer. Aromatization was determined before treatment and after 6 8 weeks on therapy by administering a bolus injection of [3H]androstenedione (500 microCi) and [14C]estrone (5 microCi) followed by measurement of the isotope ratio of urinary estrogens after high-performance liquid chromatography purification. In addition, plasma endogenous estrogens were measured with highly sensitive radioimmunoassays after separation with high-performance liquid chromatography. Treatment with exemestane suppressed whole body aromatization from a mean pretreatment value of 2.059% to 0.042% (mean suppression of 97.9%). Plasma levels of estrone, estradiol, and estrone sulfate were found to be suppressed by 94.5%, 92.2%, and 93.2%, respectively. This is the first study revealing near total aromatase inhibition in vivo with the use of a steroidal aromatase inhibitor. The observation that exemestane is a highly potent aromatase inhibitor, together with the fact that the drug is administered p.o. and causes limited side effects, suggests that exemestane is a promising new drug for the treatment of hormone sensitive breast cancer. PMID- 9748125 TI - Phase I study of a five-day dose schedule of 4-Ipomeanol in patients with non small cell lung cancer. AB - The mammalian pulmonary toxin 4-ipomeanol (IPO) is activated by the cytochrome P450 system in bronchial Clara cells in animals. The resulting metabolites bind rapidly to macromolecules, producing localized cytotoxicity. IPO has in vitro and in vivo antitumor activity in non-small cell lung cancer (NSCLC) and thus was proposed as a lung cancer-specific antitumor agent. We have completed a directed Phase I trial in patients with NSCLC. Forty-four patients (34 men and 10 women) with NSCLC were treated with IPO. All but two patients had an Eastern Cooperative Oncology Group performance status of 0 or 1. They received 91 courses of therapy with i.v. IPO; 82 courses were administered daily for five days, and 9 were single bolus doses. The dose-limiting toxicity of elevated serum transaminases was observed in three of seven patients at 922 mg/m2/day. The maximum tolerated dose was 693 mg/m2/day on 5 consecutive days every 3 weeks. One patient developed grade 4 pulmonary toxicity at 167 mg/m2/day. There was no significant hematological or renal toxicity. No objective antitumor responses were observed. Pharmacokinetic analysis of 39 patients from day 1 of IPO administration showed biexponential elimination with mean half-lives of 8.6 (alpha half-life) and 76 min (beta half-life). There was a linear relationship between the area under the plasma drug concentration-time curve and the dose of IPO. There was no significant difference between the pharmacokinetic parameters measured on day 1 and day 5. Using a 4-day in vitro cytotoxicity assay, two tumor cell lines established from patients treated at 693 mg/m2/day had IC50s of approximately 6 mM, a concentration more than 75-fold higher than the plasma levels measured in these patients. Thus, although the total amount of drug administered per cycle on a daily times five dose schedule is more than 2.5-fold higher than the recommended single daily dose, IPO is unlikely to be a useful drug for patients with lung cancer. PMID- 9748126 TI - Cell-mediated immunological responses in cervical and vaginal cancer patients immunized with a lipidated epitope of human papillomavirus type 16 E7. AB - Human papillomavirus (HPV) infection has been causally associated with cervical cancer. We tested the effectiveness of an HLA-A*0201-restricted, HPV-16 E7 lipopeptide vaccine in eliciting cellular immune responses in vivo in women with refractory cervical cancer. In a nonrandomized Phase I clinical trial, 12 women expressing the HLA-A2 allele with refractory cervical or vaginal cancer were vaccinated with four E786-93 lipopeptide inoculations at 3-week intervals. HLA-A2 subtyping was also performed, and HPV typing was assessed on tumor specimens. Induction of epitope-specific CD8+ T-lymphocyte (CTL) responses was analyzed using peripheral blood leukapheresis specimens obtained before and after vaccination. CTL specificity was measured by IFN-gamma release assay using HLA A*0201 matched target cells. Clinical responses were assessed by physical examination and radiographic images. All HLA-A*0201 patients were able to mount a cellular immune response to a control peptide. E786-93-specific CTLs were elicited in 4 of 10 evaluable HLA-A*0201 subjects before vaccination, 5 of 7 evaluable HLA-A*0201 patients after two vaccinations, and 2 of 3 evaluable HLA A*0201 cultures after all four inoculations. Two of three evaluable patients' CTLs converted from unreactive to reactive after administration of all four inoculations. There were no clinical responses or treatment toxicities. The ability to generate specific cellular immune responses is retained in patients with advanced cervical cancer. Vaccination with a lipidated HPV peptide epitope appears capable of safely augmenting CTL reactivity. Although enhancements of cellular immune responses are needed to achieve therapeutic utility in advanced cervical cancer, this approach might prove useful in treating preinvasive disease. PMID- 9748127 TI - Phase I study of the duocarmycin semisynthetic derivative KW-2189 given daily for five days every six weeks. AB - The duocarmycins represent a new group of antitumor antibiotics produced by Streptomyces that bind to the minor groove of DNA. KW-2189 is a water-soluble semisynthetic derivative of duocarmycin B2, with significant activity in murine and human tumor models. We conducted a Phase I trial of KW-2189 in patients who had solid tumors that were refractory to standard chemotherapy or for whom no more effective therapy existed. KW-2189 was administered as a rapid i.v. bolus daily for 5 days every 6 weeks. Twenty-two patients were enrolled and received a total of 31 cycles of KW-2189. Leukopenia, neutropenia, and thrombocytopenia were the dose-limiting toxicities, with nadirs occurring at medians of 36, 38, and 29 days, respectively, at the 0.04 mg/m2/day dose level. Nonhematological toxicities were mild, although one patient developed grade 3 fatigue. Four patients had stable disease over two to four cycles of treatment and showed no cumulative toxicity. The mean t1/2, plasma clearance, and steady-state volume of distribution were 13.5 min, 1,287 ml/min/m2, and 10,638 ml/m2, respectively. Pharmacokinetics were similar on days 1 and 5, with no drug accumulation in plasma. The active metabolite DU-86 was not consistently found in patient plasma. For Phase II trials, when the 5 days every 6 weeks schedule was used, 0.04 mg/m2/day KW-2189 appears to be the maximal tolerated dose, especially for patients who have received prior chemotherapy. At this dose level, the drug was well tolerated, and the toxicities were acceptable. PMID- 9748128 TI - Effect of neoadjuvant androgen deprivation on circulating prostate cells in the bone marrow of men undergoing radical prostatectomy. AB - Our objective was to determine the effect of neoadjuvant hormonal therapy on the presence of circulating prostate cells in patients undergoing radical prostatectomy for prostate cancer. A total of 60 patients at high risk for extraprostatic disease were analyzed for the presence of circulating prostate cells using reverse transcriptase PCR (RTPCR) amplification of the prostate specific antigen mRNA. Twenty-nine patients underwent radical prostatectomy for a clinical T2b-c tumor or a stage T1c-T2a tumor and a serum prostate-specific antigen level > or =10ng/ml (radical prostatectomy alone), and 31 similarly staged patients received neoadjuvant hormonal therapy before radical prostatectomy (neoadjuvant). Bone marrow samples were used for RTPCR analysis. Twenty-four percent and 58% of the radical-prostatectomy-alone patients and neoadjuvant patients had organ-confined disease, respectively (P = 0.007). In the radical-prostatectomy-alone group, 77% and 14% of patients with extraprostatic and organ-confined disease were RTPCR positive, respectively (P = 0.03). However, in the neoadjuvant group, 46% and 28% of patients with extraprostatic and organ confined disease were RTPCR positive, respectively (P = 0.29). For patients that were RTPCR positive, 45 % of the neoadjuvant patients had organ-confined disease compared with 6% in the radical-prostatectomy-alone patients (P = 0.018). These data suggest that a subset of the neoadjuvant patients are converted to organ confined disease without eliminating the prostate cells in the bone marrow. Our data suggest that hormonal therapy before radical prostatectomy decreases the occurrence of extraprostatic disease but, to a lesser degree, the incidence of circulating prostate cells. This may partially explain why hormonal therapy before radical prostatectomy has not improved disease-free survival. PMID- 9748129 TI - Changes in folate status as determined by reduction in total plasma homocysteine levels during leucovorin modulation of 5-fluorouracil therapy in cancer patients. AB - We measured plasma total homocysteine (tHcy) in 14 patients (13 patients with colorectal cancer and 1 patient with breast cancer) during their first treatment with 5-fluorouracil (5-FU) plus leucovorin [LV (5-FULV)]. Eight of these patients were investigated a second time after 3-10 cycles (median, 4 cycles) with 5-FULV. Each cycle consisted of two administrations of 5-FU (500 mg/m2) and LV (60 mg/m2) given 24 h apart. The first administration of 5-FULV on day 1 of the first cycle induced a rapid reduction of the tHcy level from 12.5 micromol/liter (10.4-15.1 micromol/liter; geometric mean with 95% confidence interval of the mean) to 9.1 micromol/liter (7.5-11.1 micromol/liter) in 24 h. tHcy remained stable at this level after the second administration of 5-FULV. In addition, the 5-FULV regimen caused a concurrent 4-fold increase in both serum and erythrocyte folate. The fifth cycle with 5-FULV had only marginal effects on the tHcy level. 5-FU without LV modulation had no effect on the plasma tHcy or folate status in eight breast cancer patients. Our data establish the reduction of tHcy as a responsive indicator of LV pharmacodynamics. PMID- 9748130 TI - Tumor angiogenesis and micrometastasis in bone marrow of patients with early gastric cancer. AB - In a subset of patients with early gastric cancer, there were recurrences of the disease after a curative resection had been done. Direct evidence of tumor seeding in distant organs at the time of surgery for gastric cancer is not available. An immunocytochemical assay for epithelial cytokeratin protein may fill this gap because it is a feature of epithelial cells that would not normally be present in bone marrow. From 1994-1997, the bone marrow of 45 patients with early gastric cancer was examined for tumor cells, using immunocytochemical techniques and an antibody reacting with cytokeratin, a component of the intracytoplasmic network of intermediate filaments. Intratumoral microvessels were stained with anti-CD31 monoclonal antibody. Clinicopathological characteristics were determined for subjects with cytokeratin-positive cells in the bone marrow. Of these 45 patients, 9 (20.0%) had cytokeratin-positive cells in the bone marrow at the time of primary surgery. These positive findings were not related to tumor advance-related factors of lymph node metastasis and distinct lymphatic and vascular invasion. Microvessel density in the primary tumor exceeded 2-fold in cytokeratin-positive cells, compared with findings in negative cells (P < 0.05). Tumor cells in bone marrow are indicative of the general disseminative metastasis in patients with early gastric cancer, and the metastatic potential was closely related to angiogenesis in the primary tumor. PMID- 9748131 TI - Disialoganglioside G(D2) loss following monoclonal antibody therapy is rare in neuroblastoma. AB - Ganglioside GD2 is abundant on human neuroblastoma (NB). Monoclonal antibody 3F8 targeted to GD2 may have imaging and therapeutic potential. Antigen-negative clones can escape immune-mediated attack, leading to clinical resistance or recurrence. Among 95 evaluable patients treated i.v. with 3F8 (94 stage 4 and 1 stage 3), 66 received nonradiolabeled 3F8, 11 received 131I-labeled 3F8 (8-28 mCi/kg) with autologous bone marrow rescue, and 18 received both forms of treatment. Prior to treatment, 91 patients tested positive for GD2 reactivity by bone marrow immunofluorescence (n = 68), tumor immunohistochemistry (n = 20), or diagnostic radioimmunoscintigraphy only (n = 3). Of 62 patients who had refractory or recurrent NB following 3F8 treatment, 61 (98%) tested positive for GD2 reactivity by bone marrow immunofluorescence (n = 51) or tumor immunohistochemistry (n = 10). The sole tumor that lost GD2 expression underwent phenotypic transformation into a pheochromocytoma-like tumor. The persistence of GD2 expression in refractory or recurrent NB suggests that complete antigen loss is an uncommon event and cannot account for treatment failure. PMID- 9748132 TI - Limitations of the reverse transcription-polymerase chain reaction method for the detection of carcinoembryonic antigen-positive tumor cells in peripheral blood. AB - To examine the limitations of reverse transcription (RT)-PCR for the detection of circulating tumor cells in blood, we established a RT-PCR for carcinoembryonic antigen (CEA). Whole blood (10(7) nucleated cells) was mixed with cells from the colon cancer cell line LS174T (concentrations ranging from 0 to 10(6) cells). RT PCR was performed to detect CEA mRNA in blood under various conditions. Healthy blood donors (n = 24) were examined by the established method for detecting CEA mRNA in blood. We were able to show that there is a detection limit for RT-PCR of 10 tumor cells in total and of 1 tumor cell in 10(5) nucleated cells. To obtain these results, a high number of PCR cycles (first PCR, 30 cycles; nested PCR, 45 cycles) was required. Under these PCR conditions, we found a positive PCR signal in 33% of healthy blood donors (n = 8). To overcome this problem, we reduced the nested PCR to 35 cycles. At that point, none of the controls showed a positive signal for CEA, and there was a subsequent decrease of the detection limit to 1 tumor cell in 10(2)-10(3) nucleated cells, lower than the detection limit of an immunocytological examination (1 tumor cell in 10(4) nucleated cells). When the amplification was performed with the tumor cells only and with no nucleated blood cells present, under exactly the same conditions, there was still a detection limit of 1 tumor cell in 106 nucleated cells. Our data clearly show that there is a severe loss of expected sensitivity of RT-PCR if it is performed in blood or nucleated blood cells. We conclude that PCR for CEA mRNA expression is not more sensitive than immunocytology and is, furthermore, plagued by the problem of a high percentage of false positive results. PMID- 9748133 TI - Circulating p53 antibodies as early markers of oral cancer: correlation with p53 alterations. AB - p53 aberrations are early events in the pathogenesis of betel- and tobacco related oral malignancies. Accumulation of p53 protein in oral lesions may elicit a humoral immune response against p53 protein in these patients. p53 antibodies (Abs) were analyzed in 183 sera obtained from patients with premalignant or malignant oral lesions and normal individuals by enzyme-linked immunoassay using recombinant p53 protein as antigen. These results were correlated with accumulation of p53 protein in patients' matched oral tissue specimens. Circulating p53 Abs were observed in 24 of 70 (34%) cancer patients and 15 of 50 (30%) patients with premalignant oral lesions. p53 Abs showed a significant association with increase in tumor size and dedifferentiation of tumors, factors indicative of poor prognosis. Expression of p53 protein was analyzed in 43 matched oral lesions (18 premalignant and 25 malignant cases). All the p53 seropositive patients (7 leukoplakia and 11 squamous cell carcinoma) showed elevated levels of p53 protein in matched oral lesions. However, the total number of patients seropositive for p53 Abs was lesser than that of patients exhibiting p53 protein accumulation in oral lesions. Four of the 63 normal healthy individuals who were heavy consumers of tobacco (smoking/chewing) and betel were found to be positive for p53 Abs. Detection of circulating p53 Abs in patients with premalignant oral lesions suggests that humoral immune response against p53 protein is an early event in oral oncogenesis and may be a surrogate marker for both p53 alteration and preclinical cancer. PMID- 9748134 TI - Specific transcription factors prognostic for prostate cancer progression. AB - We have previously identified (M. Wang et al., Oncol. Res., in press, 1998) an enhancer element [human tissue inhibitor of metalloproteinase-1 enhancer-1 (HTE)] for the human tissue inhibitor of metalloproteinase-1 promoter that binds a novel zinc finger, cysteine-rich transcription factor (CRTF). In this study, we have used electrophoretic mobility shift assays to examine the relative level of expression of CRTF, jun/fos, and IFN-gamma responsive signal transducer activators of transcription (STATs) that bind specific HTE, activator protein, and IFN-gamma (Fcy and interferon regulatory factor) response motifs in tumor lines and human prostate tissue [i.e., normal (n = 3); benign prostatic hyperplasia (BPH; n = 12); high grade prostate intraepithelial neoplasia (PIN; n = 10); and prostate cancer adenocarcinoma (PCA; n = 61) plus seminal vesicle (n = 10) tissues]. The data showed that CRTF was overexpressed in PCA (Gleason's score, 10>8>6>5>4) compared with BPH, PIN, seminal vesicle, and normal tissues. To a much lesser degree, jun/fos and STAT 1 were also elevated in PCA compared to BPH, PIN, and normal tissues. In addition, blinded studies showed that CRTF and jun/fos were present at low levels in organ-confined specimens but at significantly elevated levels (P < 0.001) in samples exhibiting capsular penetration and localized spread, which indicated that CRTF and perhaps jun/fos were markers for cancer progression. PMID- 9748135 TI - Angiogenin expression and prognosis in primary breast carcinoma. AB - Angiogenin is a protein originally isolated as an inducer of new blood vessel growth, and it has been reported to be an effective substrate for tumor cell adhesion. To understand the role of angiogenin in cancer progression, we evaluated the expression of angiogenin in 459 cases with primary breast carcinoma and in 40 benign breast specimens using an immunoassay. Higher angiogenin concentrations were observed in carcinomas in comparison with fibrocystic disease (mean, 17.3 versus 10.9 ng/mg; P = 0.008), but not with fibroadenomas. We selected 5 ng/mg cytosol protein of angiogenin as the normal cutoff for primary breast carcinoma. Eighty-eight percent of carcinomas expressed elevated angiogenin levels and 12% had low levels. We observed an association between elevated levels of angiogenin and low/ moderate histological grade (P = 0.001) and small tumor size (P = 0.026), but not with age, menopausal status, lymph node status, stage of disease, or hormonal receptor status. With a median follow-up of 31 months, breast cancer patients with elevated angiogenin levels had significantly longer disease-free survival (DFS) than patients with low angiogenin (log-rank, P = 0.003). This effect was equally observed in node negative and node-positive cases. In a multivariate analysis of DFS, only angiogenin, tumor size, and histological grade showed statistical significance. A multivariate analysis of overall survival showed that angiogenin and tumor size were the only significant variables. Serum samples from the breast cancer patients at the time of surgery were available in 194 cases. We evaluated the levels of circulating angiogenin using the same immunoassay as in tumor tissue. Serum angiogenin levels were higher in cancer patients than in 40 healthy controls (mean, 401.2 versus 206.0 ng/ml; P < 0.0001). In breast cancer patients, we observed no correlation between the serum concentrations and the tissue levels of angiogenin (r = 0.115; P = 0.110). In addition, serum levels of angiogenin did not have a prognostic impact on the DFS of breast cancer patients (log-rank, P = 0.581). Our results indicate that elevated levels of tissue angiogenin, but not of circulating angiogenin, are a favorable prognostic factor in primary breast carcinoma, which is consistent with a role of angiogenin as a cancer cell substrate. PMID- 9748137 TI - Augmented membrane type 1 matrix metalloproteinase (MT1-MMP):MMP-2 messenger RNA ratio in gastric carcinomas with poor prognosis. AB - The activation of zymogen and the amount of proteinase and its inhibition are important in determining the eventual activity of matrix-degrading enzymes involved in tumor aggressiveness. To evaluate a gene complement leading to matrix metalloproteinase 2 (MMP-2; Mr 72,000 gelatinase) activity, membrane type 1 MMP (MT1-MMP), urokinase-type plasminogen activator, MMP-2, and tissue inhibitor of metalloproteinase 2 transcriptional levels were measured in gastric carcinoma biopsies. Comparative tumor:normal tissue reverse transcription-PCR in a cohort of 25 patients revealed up to a 10-fold difference in the expression of MT1-MMP, a metalloproteinase that has been proposed as a membrane receptor activator of MMP-2; a 1-unit increment resulted in a 30% risk to survival. A 20% risk also resulted from a 1-unit increment in the MT1-MMP: MMP-2 ratio, which showed differences of up to 15-fold. Instead, the expression of urokinase-type plasminogen activator, which trips off a cascade ending in the activation of MMP 2, as well as the expression of MMP-2 itself and its inhibitor, tissue inhibitor of metalloproteinase 2, lacked correlation with patient follow-up. Zymography revealed MMP-2 activities that were often in conflict with the transcription results and also with follow-up. The results suggest the evaluation of MT1-MMP and/or MT1-MMP:MMP-2 transcription as a new preoperative molecular-level prognostic factor for gastric carcinoma. PMID- 9748136 TI - Reduced folate carrier gene expression in childhood acute lymphoblastic leukemia: relationship to immunophenotype and ploidy. AB - Reduced folate carrier (RFC) transcripts in human leukemias were measured by a competitive PCR assay. Total RNAs were reverse transcribed and amplified in the presence of competitive templates for RFC and beta-actin. RFC transcripts were normalized to transcripts for beta-actin. In a series of K562 sublines, a approximately 30-fold range of RFC transcripts measured by PCR assay closely agreed with results of Northern analysis and varied in proportion to RFC protein on Western blots and [3H]methotrexate transport. RFC transcripts varied over a 88 fold range in 49 specimens from 48 children with acute lymphoblastic leukemia (ALL). Median RFC transcripts were similar for 15 T-cell and 33 B-precursor ALL samples (RFC/beta-actin = 6.13 x 10(-3) and 7.92 x 10(-3), respectively) and for 41 diagnostic (7.20 x 10(-3)) and 8 relapse (5.58 x 10(-3)) samples. Whereas PCR measurements of RFC transcripts approximated changes in methotrexate transport in B-precursor ALL blasts (n = 10), for T-ALL blasts (n = 12) there was no apparent relationship between these parameters. For hyperdiploid B-precursor blasts (n = 11) with greater than 52 chromosomes and three to five copies of chromosome 21, the median RFC transcript level was approximately 3-fold higher than that for diploid B-precursor blasts. RFC transcripts were also elevated for two of three B precursor specimens with acquired trisomy 21. Our results suggest that RFC gene expression is far more predictive of methotrexate uptake capacity in B-precursor than T-ALL and that increased copies of chromosome 21 in B-precursor ALL blasts are generally associated with increased RFC transcripts. Hence, the good prognosis for children with hyperdiploid B-precursor ALL treated with antimetabolite-based chemotherapy and the high levels of methotrexate and methotrexate polyglutamates accumulated may, in part, reflect elevated RFC gene expression and capacities for methotrexate transport. PMID- 9748138 TI - Expression profile of prostate-specific antigen messenger RNA assessed by in situ hybridization is a novel prognostic marker for patients with untreated prostate cancer. AB - The present study was undertaken to define the relationship between histological grade (Gleason grade) and prostate-specific antigen (PSA) mRNA expression and to evaluate the level of PSA mRNA expression as a possible prognostic marker for untreated prostate cancers. The primary grade areas of 104 prostatic biopsy specimens were analyzed for the expression of PSA mRNA and its protein by nonradioactive in situ hybridization and immunohistochemistry, respectively. A multivariate survival analysis was performed to examine the correlation between PSA mRNA expression and several clinicopathological parameters, e.g., the immunostaining level of PSA protein in biopsy specimens. The percentage of specimens positive for PSA mRNA increased significantly with advanced histological grade. Image analysis of the signal intensity for PSA mRNA showed a significant correlation between the signal intensity in both primary and secondary grade areas of each specimen and the histological grade (P < 0.0001). Only 26.0% of specimens positive for PSA protein were also positive for PSA mRNA (and vice versa, 6.7%). Other tumors were either positive for both (66.3%) or negative for both (1.0%). When the Cox's proportional hazards regression model was used to analyze cancer-specific survival, untreated patients with higher levels of PSA mRNA expression in the higher grade (representing higher grade of either primary or secondary grade) area of tumors were at high risk for cancer related death (P = 0.017). Furthermore, in cancer-specific survival curves based on PSA mRNA expression status, patients with high levels of PSA mRNA expression in the higher grade area of tumors had a significantly poorer prognosis (P = 0.001), compared with those with tumors expressing low levels of PSA mRNA. Our results suggested that analysis of PSA mRNA expression in specific areas in biopsy specimens of patients with untreated prostate cancer may provide a good assessment of prognosis of prostate cancers. PMID- 9748139 TI - Significance of vessel count and vascular endothelial growth factor in human esophageal carcinomas. AB - The purpose of this study was to determine the angiogenic profile of human esophageal carcinomas. The expression of vascular endothelial growth factor (VEGF) was examined in 6 esophageal carcinoma cell lines and 119 human esophageal carcinoma tissues by Northern blot analysis and immunohistochemistry, respectively. Immunohistochemistry using antibodies against CD34 (endothelial cell specific) was carried out on archival specimens, and microvessels were quantitated by counting vessels in a x200 field in the most vascular area of the tumor. All of the cell lines constitutively expressed VEGF mRNA at various levels. A total of 71 of 119 (59.7%) tumors showed intense VEGF immunoreactivity in the cytoplasm of cancer cells. Vessel count was significantly higher in the VEGF-positive tumors than it was in the VEGF-negative tumors. VEGF expression correlated with the depth of tumor invasion, tumor stage, venous invasion, and lymphatic invasion. The survival rate of patients with high vessel density in the tumor was significantly worse than that of patients with low vessel density in the tumor. There was a tendency for poorer prognosis in the group with VEGF positive tumors compared with that of the group with VEGF-negative tumors. Overall, these results suggest that VEGF is associated with tumor progression by stimulating angiogenesis in human esophageal carcinoma. PMID- 9748140 TI - UCN-01 suppresses thymidylate synthase gene expression and enhances 5 fluorouracil-induced apoptosis in a sequence-dependent manner. AB - UCN-01, a protein kinase C/cyclin-dependent kinase inhibitor, suppressed thymidylate synthase (TS) protein expression in a dose-dependent manner with near complete suppression at 1 microM after a 24-h exposure in human gastric cancer cell line SK-GT5. Other protein kinase C/cyclin-dependent kinase inhibitors, including flavopiridol and safingol, had a similar effect on TS protein expression, but to a lesser degree. Moreover, UCN-01 repressed the induction of TS after 5-fluorouracil (FU) exposure by 90-95% and significantly enhanced the induction of apoptosis by FU from 4-8% with either FU or UCN-01 alone to 46+/-1% (P < 0.005 versus either single drug, reverse sequence, or the combination) when UCN-01 was given after FU. The effect of UCN-01 on TS was associated with a dose dependent suppression of the E2F-1 protein, a transcriptional activator of TS. Northern blot analysis revealed that TS mRNA levels decreased gradually as the concentration of UCN-01 increased, but that E2F-1 mRNA levels remained relatively unchanged. UCN-01 may provide a novel way to enhance cellular sensitivity toward FU by means of suppressing TS expression mediated mainly by down-regulation of E2F-1. PMID- 9748141 TI - Photodynamic therapy of naturally occurring tumors in animals using a novel benzophenothiazine photosensitizer. AB - 5-Ethylamino-9-diethylaminobenzo[a]phenothiazinium chloride (EtNBS) is a novel photodynamic therapy (PDT) photosensitizer with efficacy against experimental murine tumors. In this preliminary study, dogs and cats with naturally occurring tumors were treated with EtNBS-PDT to determine safety and efficacy. Fifteen treatments were performed on 13 animals (9 treatments in 8 cats and 6 treatments in 5 dogs), generally using 400 J of 652 nm light. Two feline sublingual squamous cell carcinomas (SCCs) responded briefly (minor response). Six feline facial SCCs were treated, resulting in two partial responses and four long-term complete responses (CR). Two canine intraoral SCCs were treated; one responded minimally for 2 weeks (minor response), and one achieved long-term CR. One canine cutaneous mast cell tumor achieved CR, and one canine ocular mast cell tumor responded briefly. One canine ocular melanoma did not respond to treatment. Systemic reactions included nausea associated with photosensitizer injection in two cats and two dogs, elevated body temperatures during treatment in two dogs, elevated body temperature 2 days after PDT in one cat, and inappetance for 2 weeks in one cat. A peripheral neuropathy of undetermined cause occurred in one cat 2 weeks after PDT and resolved without treatment. Local reaction was well tolerated in 13 of 15 treatments. All animals were exposed to normal daylight after less than 5 days (mean, 3.5 days) without residual photosensitization. EtNBS-PDT is safe for dogs and cats and has activity against selected naturally occurring tumors, with an overall objective response rate (partial response + CR) of 61.5%. PMID- 9748143 TI - Desensitization and sensitization of cells to fluoropyrimidines with different antisenses directed against thymidylate synthase messenger RNA. AB - Previous studies have shown that the cytotoxicity of fluoropyrimidines is mediated, in large part, by inhibition of the enzyme thymidylate synthase (TS). The aim of this study was to determine whether the chemosensitivity of human cancer cells to fluoropyrimidines could be increased by decreasing TS expression with antisense oligodeoxyribonucleotides (ODNs). ODNs (18-mers) targeted at the AUG translational initiation site of TS mRNA inhibited translation in a sequence- and dose-dependent manner in a rabbit reticulocyte lysate in vitro translation system. Treatment of human colon cancer HT-29 cells with antisense ODNs decreased TS catalytic activity in the cells in a dose-dependent manner over a short period, but the longer-term effect of the TS antisense ODN treatment was actually to increase the amount of TS in the cells and to decrease their sensitivity to 5 fluoro-2'-deoxyuridine (FdUrd). However, when human nasopharyngeal cancer KB31 cells were transfected with a plasmid (pHaMAGRP) construct containing the TS antisense fragment (+ 1 to + 422) under the control of a glucose-regulated promoter, the expression of both TS protein and TS catalytic activity was decreased by nearly 30% (P = 0.014), and sensitivity of these cells to FdUrd was enhanced by approximately 8-fold (P = 0.021). No changes in the levels of expression of TS protein or FdUrd-associated cytotoxicity were observed in control, vector-transfected cells. No change was observed in the sensitivity of transfected cells toward either cisplatin or Adriamycin. These results show that the level of expression of TS in human malignant cells can be down-regulated with antisense TS RNA, and their sensitivity to fluoropyrimidines can, thereby, be increased. PMID- 9748142 TI - Interferon-alpha-induced activation of signal transducer and activator of transcription proteins in malignant melanoma. AB - IFN-alpha2b has been used to treat patients with malignant melanoma who are at high risk for recurrence after surgical resection. However, its exact mechanism of action is unknown. I hypothesized that IFN-alpha exerts a direct effect on the melanoma cell via the activation of signal transducer and activator of transcription (STAT) proteins. Cell lysates from melanoma cell lines and patient tumor samples stimulated with IFN-alpha were incubated with radiolabeled oligonucleotides representing the high affinity sis-inducible element of c-fos and the IFN stimulated response element and then analyzed for STAT activation using the electrophoretic mobility shift assay. Melanoma cell lines showed no evidence for constitutive STAT activation in the absence of cytokine stimulation but exhibited rapid activation of STAT1 and STAT2 once treated with IFN-alpha. A distinct dose-response curve was noted with maximal STAT activation occurring at approximately 10(5) units/ml IFN-alpha. Genistein, a protein tyrosine kinase inhibitor, completely suppressed IFN-alpha-induced STAT activation. Malignant melanoma tumors obtained from 17 patients exhibited dose-dependent activation of STAT1 and STAT2 in response to treatment with IFN-alpha. Pretreatment of patient melanoma tumor cells with IFN-gamma resulted in a 4 log-fold decrease in the IFN alpha concentration required for STAT activation and promoted the increased expression of STAT1, STAT2, and p48. In summary, IFN-alpha consistently activated STAT1 and STAT2 proteins in melanoma cell lines and in melanoma tumors obtained directly from patients. This signaling pathway was dramatically sensitized to IFN alpha by pretreatment of melanoma cells with low concentrations of IFN-gamma. These results provide molecular evidence to support the hypothesis that the clinical response to IFN-alpha may be mediated, in part, by a direct effect on the melanoma cell. These results also suggest a potential role for IFN-gamma in the treatment of malignant melanoma. PMID- 9748144 TI - Characterization of a novel bispecific antibody that mediates Fcgamma receptor type I-dependent killing of tumor-associated glycoprotein-72-expressing tumor cells. AB - A bispecific antibody was made by chemical conjugation of Fab' fragments from humanized antibodies specific for tumor-associated glycoprotein-72 (TAG-72) and high-affinity immunoglobulin receptor, FcgammaA receptor type I (FcgammaRI). The purified anti-TAG-72 x anti-FcgammaRI (HCC49xH22) bispecific antibody had an approximate Mr of 111,000, consistent with a F(ab')2, and bound specifically to KLEB and LS174T tumor cell lines, which express the TAG-72 tumor antigen. Furthermore, HCC49x H22 was shown to simultaneously bind to KLEB cells and a soluble FcgammaRI fusion protein, demonstrating the bifunctional nature of the molecule. Using IFN-gamma-treated monocytes as effector cells, concentrations of the bispecific antibody in the range of 1-10,000 ng/ml mediated specific lysis of TAG-72-positive tumor cells. In contrast, the bispecific antibody did not promote antibody-dependent cellular cytotoxicity of a cell line that was negative for TAG 72 antigen. Importantly, the antibody-dependent cellular cytotoxicity activity of the bispecific antibody was significantly greater than that of the monoclonal antibody HCC49. These in vitro data indicate that the humanized bispecific antibody HCC49xH22 has the appropriate specificity and functional activity for further evaluation as potential immunotherapy for TAG-72-positive malignancies. PMID- 9748145 TI - Functionality of the progesterone receptor in ovarian cancer and its regulation by estrogen. AB - Here, we sought to obtain evidence that the progesterone receptor (PR) may be functional in ovarian cancer and regulated by estrogen. Megestrol acetate inhibited growth of the PR-positive PE04 ovarian carcinoma xenograft but not the PR-negative HOX 60 xenograft. PR concentration was higher in early-stage (I/II) tumors than in advanced-stage (III/IV) tumors (P = 0.007) and in tumors of endometrioid histology compared to other carcinoma subtypes (P = 0.009). Patients with a tumor PR concentration of >40 fmol/mg protein had significantly improved survival over those patients whose tumors contained <40 fmol/mg (P = 0.0007; log rank). Evidence of PR regulation by estrogen was obtained by endocrine manipulation of the PE04 xenograft. PR content of PE04 xenografts fell from 145 to 7 fmol/mg protein in ovariectomized mice and was 2 fmol/mg in male mice. Administration of 17-beta-estradiol increased PR content to 745 fmol/mg. In primary ovarian carcinomas, PR was significantly associated with ER concentrations (P < 0.0001), suggesting regulation of PR levels by estrogen. This association was present for tumors of endometrioid histology (P < 0.0001) but not for those with serous histology (P = 0.31). These data point to the regulation of PR levels by estrogen in ovarian cancer and to a mediatory role for PR in the inhibition of growth induced by progestin. PMID- 9748146 TI - Toxicity and dose-response studies of 1,25-(OH)2-16-ene-23-yne vitamin D3 in transgenic mice. AB - The vitamin D3 analogue 1,25-(OH)2-16-ene-23-yne vitamin D3 (16,23-D3) in doses with low systemic toxicity has been demonstrated to inhibit retinoblastoma growth in transgenic mice. This study examines the dose-dependent response for inhibition of tumor growth in transgenic mice with retinoblastoma and evaluates the in vivo toxicity of 16,23-D3 in nontransgenic mice. Transgenic 8-10-week-old mice with retinoblastoma (n = 119) were randomly assigned to groups receiving 1.0, 0.75, 0.5, 0.35, 0.2, or 0.05 microg of 16,23-D3 and a vehicle alone (control) group i.p. five times a week for 5 weeks. An additional control group received no injection. Eyes were enucleated one week after the end of treatment, and tumor areas were measured. To determine the toxic dose, transgene-negative littermates received 0.5, 1.0, 1.5, 2.5, 3.5, 4.5, or 5.0 microg of 16,23-D3, and control groups received vehicle alone, 5 days a week for 5 weeks. Serum calcium levels were measured, and necropsies were performed on animals from each group. In the dose-response study, tumor growth inhibition was greatest in the group receiving 0.35 microg (55% inhibition; P = 0.0056) and was also significant in the group receiving 0.5 microg (42% inhibition; P = 0.036). The systemic toxic effects due to hypercalcemia occurred at doses of > or =1.0 microg. 16,23-D3 inhibits tumor growth at doses > or =0.35 microg and shows toxic effects at doses > or =1.0 microg related to hypercalcemia in mice fed an unrestricted diet. No toxicity was observed with lower doses. PMID- 9748147 TI - Antimestatic and antitumor activities of interleukin 10 in transfected human prostate PC-3 ML clones: Orthotopic growth in severe combined immunodeficient mice. AB - We have permanently transfected human prostate PC-3 ML tumor cells and examined the influence of interleukin 10 (IL-10) production on tumor growth and metastasis following orthotopic implantation in the prostate gland of severe combined immunodeficient mice. Measurements of tumor volume after 5, 8, and 12 weeks indicated that tumor volume was negatively correlated with the amount of IL-10 production. Likewise, the extent of metastasis was inversely related to the amount of IL-10 produced. Following i.v. injection, the IL-10-expressing clones also failed to metastasize to the bone marrow. Controls showed that PC-3 ML and PC-3 ML mock clones grew rapidly and metastasized when implanted orthotopically or injected i.v. via the tail vein. Mouse survival curves showed that all of the mice injected orthotopically with the PC-3 ML clones died by about 14-16 weeks, whereas the PC-3 ML-IL10a or PC-3 ML-ILl0b clones induced only 10-20% death after 23-24 weeks. Likewise, survival studies showed a high death rate by approximately 30 days with PC-3 ML mock cells but <10% death by 12 weeks with the IL-10 transfected clones injected i.v. via the tail vein. The data strongly suggest that IL-10 production blocks tumor growth and metastasis in severe combined immunodeficient mice. PMID- 9748148 TI - Combination E2F-1 and p53 gene transfer does not enhance growth inhibition in human squamous cell carcinoma of the head and neck. AB - Ample data exist contending that wild-type p53 and E2F-1 cooperate to mediate apoptosis, that E2F-1-mediated apoptosis is p53 dependent in some situations, and that E2F-1 can induce accumulation of p53 in mammalian cells. These data support the investigation of the biological consequences of combined wild-typep53 and E2F 1 overexpression in human squamous cell carcinoma of the head and neck (SCCHN) for the purpose of developing apoptosis-inducing molecular intervention strategies for the management of this devastating disease. The recombinant adenovirus (Ad) vectors Ad-p53 and Ad-E2F-1 were used for wild-type p53 and E2F-1 gene transfers, respectively, into SCCHN cell lines TU138 and TU167. SCCHN cells transduced with either p53, E2F-1, or both underwent in vitro growth analysis, which revealed that simultaneous p53 and E2F-1 gene transfer did not result in enhanced growth inhibition. To explain our growth assay findings on the basis of potential negative molecular interactions between E2F-1 and p53, Western and Northern blotting analyses were performed to investigate the differential expression of the downstream p53-transactivated genes, p21Waf1 and BAX, under various p53 and E2F-1 gene transfer conditions. Whereas Western immunoblotting demonstrated that E2F-1 antagonized p53 induction of p21Waf1 and BAX, Northern blotting revealed that this interference was pretranslationally regulated and p53 dependent. Coimmunoprecipitation assay confirmed that the wild-type p53 and E2F-1 gene products formed protein-protein complexes in our cell lines. Our in vitro data demonstrated that in SCCHN, E2F-1 interferes with induction of p53 transactivated genes, probably through the formation of protein-protein complexes. Simultaneous p53 and E2F-1 gene transfer is not therapeutically advantageous in this in vitro model of SCCHN. PMID- 9748149 TI - Nuclear expression of YB-1 protein correlates with P-glycoprotein expression in human osteosarcoma. AB - The Y-box-binding protein, YB-1, is a member of the DNA-binding protein family. It binds to the Y-box, an inverted CCAAT box, in the promoter region of the human multidrug resistance 1 gene, which encodes P-glycoprotein (P-gp). Nuclear localization of YB-1 protein has been reported to be associated with the intrinsic expression of P-gp in human breast cancer. We studied the immunohistochemical expression of YB-1 protein in 69 untreated biopsy specimens of conventional osteosarcomas and compared it with the expression of P-gp. Furthermore, cell proliferation, as determined by the MIB-1-labeling index (MIB-1 LI), was measured by immunohistochemistry. In all 69 untreated osteosarcomas, YB 1 protein was expressed in the cytoplasm. In 32 of 69 (46%) cases, YB-1 was also localized in the nucleus. The expression of P-gp was evident in 23 of these 32 cases, and there was a significant correlation between the nuclear expression of YB-1 and P-gp expression (P < 0.0001). Chondroblastic osteosarcoma expressed YB-1 in the nucleus more frequently (eight of nine cases) than did other types of osteosarcoma, whereas P-gp was also frequently expressed in chondroblastic subtype. There was no correlation between the nuclear expression of YB-1 and histological grade. The MIB-1-LI was significantly higher in cases showing the nuclear expression of YB-1 (MIB-1-LI averaged 22.56 in cases with only cytoplasmic expression of YB-1 but averaged 28.20 in cases with cytoplasmic and nuclear expression of YB-1; P = 0.0477). In human osteosarcoma, nuclear localization of YB-1 protein was associated with the expression of P-gp, suggesting that YB-1 could be a prognostic marker for multidrug resistance in osteosarcoma. PMID- 9748150 TI - Immunohistochemical detection of multidrug resistance protein in human lung cancer and normal lung. AB - Monoclonal antibody QCRL-1 is highly specific for a defined linear epitope in a relatively poorly conserved region of the human multidrug resistance protein (MRP). We have used QCRL-1 to examine MRP expression in archival and fresh snap frozen samples of untreated small cell (SC) and non-small cell (NSC) lung cancers (LCs), as well as normal lung. We found that the majority (87%) of all histological subtypes of NSCLC had detectable levels of MRP in most of the tumor mass. In a substantial proportion of adenocarcinomas (55%) and squamous cell carcinomas (28%), immunoreactivity approached that obtained with the highly multidrug resistant cell line H69AR from which the MRP was originally cloned. Both the level and frequency of MRP expression in untreated SCLC was significantly lower than in NSCLC. The MRP was detectable in only 56% of SCLC tumors and, in most cases, was expressed in small focal clusters of cells. Immunofluorescence studies of tumor tissue and normal lung confirmed the plasma membrane location of the MRP. However, in normal bronchial epithelium and seromucous glands, unlike in tumor cells, the MRP was detected only on basolateral membranes. In addition, strong MRP immunoreactivity was detected in reactive type II pneumocytes present in hyperplastic alveoli, but not in normal type I and type II pneumocytes. No potentially confounding correlation independent of its possible role in drug resistance was observed between MRP expression in untreated NSCLC and any clinicopathological parameter examined, including overall survival. PMID- 9748151 TI - Why sex and recombination? AB - REVIEW Most higher organisms reproduce sexually, despite the automatic reproductive advantage experienced by asexual variants. This implies the operation of selective forces that confer an advantage to sexuality and genetic recombination, at either the population or individual level. The effect of sex and recombination in breaking down negative correlations between favorable variants at different genetic loci, which increases the efficiency of natural selection, is likely to be a major factor favoring their evolution and maintenance. Various processes that can cause such an effect have been studied theoretically. It has, however, so far proved hard to discriminate among them empirically. PMID- 9748152 TI - The evolutionary dynamics of sex determination. AB - REVIEW There is substantial cytogenetic data indicating that the process of sex determination can evolve relatively rapidly. However, recent molecular studies on the evolution of the regulatory genes that control sex determination in the insect Drosophila melanogaster, the nematode Caenorhabditis elegans, and mammals suggest that, although certain sex determination regulatory genes have evolved relatively rapidly, other sex determination regulatory genes are quite conserved. Thus, studies of the evolution of sex determination, a process that appears to have elements that undergo substantial evolutionary change and others that may be conserved, could provide substantial insights into the kinds of forces that both drive and constrain the evolution of developmental hierarchies. PMID- 9748153 TI - Evolution of gamete recognition proteins. AB - REVIEW Although fertilization has been studied for more than a century, the cell surface proteins mediating the process are only now becoming known. Gamete interaction in animals appears to be molecularly complex. Although it is difficult to generalize at present, diversity of structure may be a recurring theme in the evolution of fertilization proteins. Examples of rapid evolution of fertilization proteins by positive selection are known, and concerted evolution can influence the differentiation of gamete recognition proteins between closely related species. PMID- 9748154 TI - Sexual selection, receiver biases, and the evolution of sex differences. AB - REVIEW Recent approaches to analyzing the evolution of female mating preferences emphasize how historical influences on female receiver systems can bias the evolution of male traits that females find attractive. These studies combine animal behavior, sensory biology, phylogenetics, and artificial neural network models. They attempt to understand why specific phenotypes involved in sexual selection have evolved, rather than merely determining whether such traits and preferences are adaptive. It is now clear that traits and preferences often do not coevolve via genetic correlations, that female mating preferences for a given male trait are influenced by adaptations and constraints outside of the context of female responses to that particular trait, and that receiver biases can explain much of the diversity in male signaling phenotypes. It also appears that an understanding of historical effects will prove valuable in investigating why neural and cognitive systems respond to sensory stimuli as they do. PMID- 9748155 TI - Sex and conflict. AB - Evolutionary conflict occurs when the deterministic spread of an allele lowers the fitness either of its bearer or of other individuals in the population, leading to selection for suppressors. Sex promotes conflict because associations between alleles are temporary. Differing selection on males and females, sexual selection, and differences in transmission patterns between classes of nuclear and cytoplasmic genes can all give rise to conflict. Inert Y chromosomes, uniparental inheritance of cytoplasmic genes, mating strains and sexes, and many features of sexual behavior may have evolved in part as a result of evolutionary conflict. Estimates of its quantitative importance, however, are still needed. PMID- 9748156 TI - Ancient mantle in a modern arc: osmium isotopes in izu-bonin-mariana forearc peridotites AB - Mantle peridotites drilled from the Izu-Bonin-Mariana forearc have unradiogenic 187Os/188Os ratios (0.1193 to 0.1273), which give Proterozoic model ages of 820 to 1230 million years ago. If these peridotites are residues from magmatism during the initiation of subduction 40 to 48 million years ago, then the mantle that melted was much more depleted in incompatible elements than the source of mid-ocean ridge basalts (MORB). This result indicates that osmium isotopes record information about ancient melting events in the convecting upper mantle not recorded by incompatible lithophile isotope tracers. Subduction zones may be a graveyard for ancient depleted mantle material, and portions of the convecting upper mantle may be less radiogenic in osmium isotopes than previously recognized. PMID- 9748157 TI - Semiconductor nanocrystals as fluorescent biological labels. AB - Semiconductor nanocrystals were prepared for use as fluorescent probes in biological staining and diagnostics. Compared with conventional fluorophores, the nanocrystals have a narrow, tunable, symmetric emission spectrum and are photochemically stable. The advantages of the broad, continuous excitation spectrum were demonstrated in a dual-emission, single-excitation labeling experiment on mouse fibroblasts. These nanocrystal probes are thus complementary and in some cases may be superior to existing fluorophores. PMID- 9748158 TI - Quantum dot bioconjugates for ultrasensitive nonisotopic detection. AB - Highly luminescent semiconductor quantum dots (zinc sulfide-capped cadmium selenide) have been covalently coupled to biomolecules for use in ultrasensitive biological detection. In comparison with organic dyes such as rhodamine, this class of luminescent labels is 20 times as bright, 100 times as stable against photobleaching, and one-third as wide in spectral linewidth. These nanometer sized conjugates are water-soluble and biocompatible. Quantum dots that were labeled with the protein transferrin underwent receptor-mediated endocytosis in cultured HeLa cells, and those dots that were labeled with immunomolecules recognized specific antibodies or antigens. PMID- 9748160 TI - Origin of multikilometer earth- and mars-crossing asteroids: A quantitative simulation AB - Orbital dynamic simulations show that many asteroids in the main asteroid belt are driven toward Mars-crossing orbits by numerous weak mean motion resonances, which slowly increase the orbital ellipticity of the asteroids. In addition, half of the Mars-crossing asteroids (MCAs) transition to Earth-crossing asteroids (ECAs) in less than 20 million years. This scenario quantitatively explains the observed number of large ECAs and MCAs. PMID- 9748159 TI - Europa's differentiated internal structure: inferences from four Galileo encounters. AB - Radio Doppler data from four encounters of the Galileo spacecraft with the jovian moon Europa have been used to refine models of Europa's interior. Europa is most likely differentiated into a metallic core surrounded by a rock mantle and a water ice-liquid outer shell, but the data cannot eliminate the possibility of a uniform mixture of dense silicate and metal beneath the water ice-liquid shell. The size of a metallic core is uncertain because of its unknown composition, but it could be as large as about 50 percent of Europa's radius. The thickness of Europa's outer shell of water ice-liquid must lie in the range of about 80 to 170 kilometers. PMID- 9748161 TI - The formation of substellar objects induced by the collision of protostellar disks AB - Simulations of a close encounter between two protostars, each surrounded by a relatively massive disk, resulted in the ejection of some of the disk material into a tidal tail. A portion of the tail condensed into an object with a mass in the range of 5 to 10 jovian masses. This mechanism may explain the existence of the single objects of substellar mass that have recently been discovered. PMID- 9748162 TI - Bax and adenine nucleotide translocator cooperate in the mitochondrial control of apoptosis. AB - The proapoptotic Bax protein induces cell death by acting on mitochondria. Bax binds to the permeability transition pore complex (PTPC), a composite proteaceous channel that is involved in the regulation of mitochondrial membrane permeability. Immunodepletion of Bax from PTPC or purification of PTPC from Bax deficient mice yielded a PTPC that could not permeabilize membranes in response to atractyloside, a proapoptotic ligand of the adenine nucleotide translocator (ANT). Bax and ANT coimmunoprecipitated and interacted in the yeast two-hybrid system. Ectopic expression of Bax induced cell death in wild-type but not in ANT deficient yeast. Recombinant Bax and purified ANT, but neither of them alone, efficiently formed atractyloside-responsive channels in artificial membranes. Hence, the proapoptotic molecule Bax and the constitutive mitochondrial protein ANT cooperate within the PTPC to increase mitochondrial membrane permeability and to trigger cell death. PMID- 9748163 TI - Dorsal-ventral signaling in the Drosophila eye. AB - The development of the Drosophila eye has served as a model system for investigations of tissue patterning and cell-cell communication; however, early eye development has not been well understood. The results presented here indicate that specialized cells are established along the dorsal-ventral midline of the developing eye by Notch-mediated signaling between dorsal and ventral cells, and that Notch activation at the midline plays an essential role both in promoting the growth of the eye primordia and in regulating eye patterning. These observations imply that the developmental homology between Drosophila wings and vertebrate limbs extends to Drosophila eyes. PMID- 9748164 TI - The evolution of agriculture in ants AB - Cultivation of fungi for food by fungus-growing ants (Attini: Formicidae) originated about 50 million years ago. The subsequent evolutionary history of this agricultural symbiosis was inferred from phylogenetic and population-genetic patterns of 553 cultivars isolated from gardens of "primitive" fungus-growing ants. These patterns indicate that fungus-growing ants succeeded at domesticating multiple cultivars, that the ants are capable of switching to novel cultivars, that single ant species farm a diversity of cultivars, and that cultivars are shared occasionally between distantly related ant species, probably by lateral transfer between ant colonies. PMID- 9748165 TI - Impaired spatial learning after saturation of long-term potentiation. AB - If information is stored as activity-driven increases in synaptic weights in the hippocampal formation, saturation of hippocampal long-term potentiation (LTP) should impair learning. Here, rats in which one hippocampus had been lesioned were implanted with a multielectrode stimulating array across and into the angular bundle afferent to the other hippocampus. Repeated cross-bundle tetanization caused cumulative potentiation. Residual synaptic plasticity was assessed by tetanizing a naive test electrode in the center of the bundle. Spatial learning was disrupted in animals with no residual LTP (<10 percent) but not in animals that were capable of further potentiation. Thus, saturation of hippocampal LTP impairs spatial learning. PMID- 9748166 TI - Protein kinase C isotypes controlled by phosphoinositide 3-kinase through the protein kinase PDK1. AB - Phosphorylation sites in members of the protein kinase A (PKA), PKG, and PKC kinase subfamily are conserved. Thus, the PKB kinase PDK1 may be responsible for the phosphorylation of PKC isotypes. PDK1 phosphorylated the activation loop sites of PKCzeta and PKCdelta in vitro and in a phosphoinositide 3-kinase (PI 3 kinase)-dependent manner in vivo in human embryonic kidney (293) cells. All members of the PKC family tested formed complexes with PDK1. PDK1-dependent phosphorylation of PKCdelta in vitro was stimulated by combined PKC and PDK1 activators. The activation loop phosphorylation of PKCdelta in response to serum stimulation of cells was PI 3-kinase-dependent and was enhanced by PDK1 coexpression. PMID- 9748167 TI - Metapopulation dynamics, abundance, and distribution in a microecosystem AB - The experimental fragmentation of landscapes of a natural ecosystem resulted in declines in the abundance and distribution of most species in the multispecies animal community inhabiting the landscapes and the extinction of many species. These declines caused the deterioration of the positive interspecific relation between local population abundance and distributional extent in this community. However, when patches were connected by habitat corridors, an immigration "rescue effect" arrested declines in both abundance and distribution and maintained the observed positive relation between them. These results demonstrate the importance of metapopulation dynamics and landscape connectivity for the persistence of populations in fragmented landscapes. PMID- 9748168 TI - Alcohol and cancer. Still no clear evidence to link specific beverages to specific cancers. PMID- 9748169 TI - Cyclo-oxygenase 2 and breast cancer prevention. Non-steroidal anti-inflammatory agents are worth testing in breast cancer. PMID- 9748170 TI - Albumin: don't confuse us with the facts. Rather than fulminating, seek to answer the questions raised. PMID- 9748171 TI - Flushing away the fat. Weight loss during trials of orlistat was significant, but over half was due to diet. PMID- 9748172 TI - US managed care: has the UK anything to learn? Fishbowl medicine is here to stay. PMID- 9748173 TI - Where's the chief knowledge officer? To manage the most precious resource of all. PMID- 9748174 TI - Benzodiazepine use in pregnancy and major malformations or oral cleft: meta analysis of cohort and case-control studies. AB - OBJECTIVE: To determine if exposure to benzodiazepines during the first trimester of pregnancy increases risk of major malformations or cleft lip or palate. DESIGN: Meta-analysis. SETTING: Studies from 1966 to present. SUBJECTS: Studies were located with Medline, Embase, Reprotox, and from references of textbooks, reviews, and included articles. Included studies were original, concurrently controlled studies in any language. INTERVENTIONS: Data extraction and quality assessment were done independently and in duplicate. MAIN OUTCOME MEASURES: Maternal exposure to benzodiazepines in at least the first trimester; incidence of major malformations or oral cleft alone, measured as odds ratios and 95% confidence intervals with a random effects model. RESULTS: Of over 1400 studies reviewed, 74 were retrieved and 23 included. In the analysis of cohort studies fetal exposure to benzodiazepine was not associated with major malformations (odds ratio 0.90; 95% confidence interval 0.61 to 1. 35) or oral cleft (1.19; 0.34 to 4.15). Analysis of case-control studies showed an association between exposure to benzodiazepines and development of major malformations (3.01; 1.32 to 6.84) or oral cleft alone (1.79; 1.13 to 2.82). CONCLUSIONS: Pooled data from cohort studies showed no association between fetal exposure to benzodiazepines and the risk of major malformations or oral cleft. On the basis of pooled data from case-control studies, however, there was a significant increased risk for major malformations or oral cleft alone. Until more research is reported, level 2 ultrasonography should be used to rule out visible forms of cleft lip. PMID- 9748175 TI - Population based cohort study of the association between alcohol intake and cancer of the upper digestive tract. AB - OBJECTIVE: To examine the relation between different types of alcoholic drinks and upper digestive tract cancers (oropharyngeal and oesophageal). DESIGN: Population based study with baseline assessment of intake of beer, wine, and spirits, smoking habits, educational level, and 2-19 years' follow up on risk of upper digestive tract cancer. SETTING: Denmark. SUBJECTS: 15 117 men and 13 063 women aged 20 to 98 years. MAIN OUTCOME MEASURE: Number and time of identification of incident upper digestive tract cancer during follow up. RESULTS: During a mean follow up of 13.5 years, 156 subjects developed upper digestive tract cancer. Compared with non-drinkers (drinkers of <1 drink/week), subjects who drank 7-21 beers or spirits a week but no wine were at a risk of 3.0 (95% confidence interval 1.5 to 6.1), whereas those who had the same total alcohol intake but with wine as >=30% of their intake had a risk of 0.5 (0.2 to 1.4). Drinkers of >21 beers and spirits but no wine had a relative risk of 5.2 (2.7 to 10.2) compared with non-drinkers, whereas those who drank the same amount, but included wine in their alcohol intake, had a relative risk of 1.7 (0.6 to 4. 4). CONCLUSION: A moderate intake of wine probably does not increase the risk of upper digestive tract cancer, whereas a moderate intake of beer or spirits increases the risk considerably. PMID- 9748176 TI - Population based case-control study of sick leave in postmenopausal women before diagnosis of hyperparathyroidism. AB - OBJECTIVE: To analyse sick leave in women at risk of primary hyperparathyroidism before its diagnosis. DESIGN: Case-control study nested within a screened cohort of postmenopausal women. Cases were women with hyperparathyroidism without prior knowledge of their disease and no traditional symptoms or complications. Controls were women from the screened population without hyperparathyroidism. SETTING: Population based screening within a Swedish community. SUBJECT: 48 case-control pairs of women aged 55-70 years. MAIN OUTCOME MEASURE: Sick leave during the 5 years before diagnosis. RESULTS: Total duration of sickness benefits was longer in the cases than controls, and this discrepancy included sick leave on full time or half time and for periods of longer than a week. Cases had an increased risk of sick leave more than half of the investigated time compared with controls (odds ratio 12). Doctors' certificates showed that the overrepresented sick leave in the cases related mainly to cardiovascular diseases. CONCLUSION: Asymptomatic mild primary hyperparathyroidism in postmenopausal women is accompanied by a previously unrecognised morbidity, which has consequences for clinical management of the disorder and its impact on the health economy. PMID- 9748177 TI - Long term relative survival after surgery for abdominal aortic aneurysm in western Australia: population based study. AB - OBJECTIVE: To determine the long term relative survival of all patients who had surgery for abdominal aortic aneurysm in Western Australia during 1985-94. DESIGN: Population based study. SETTING: Western Australia. SUBJECTS: All patients who had had surgery for abdominal aortic aneurysm in Western Australia during 1985-94. MAIN OUTCOME MEASURES: Morbidity and mortality data of patients admitted and surgically treated for abdominal aortic aneurysm in Western Australia during 1985-94. Elective, ruptured, and acute non-ruptured cases were analysed separately. Independent analyses for sex and patients aged 80 years or more were also undertaken. Postoperative (>30 days) relative survival was assessed against age and sex matched controls. RESULTS: Overall, 1475 (1257 men, 218 women) cases were identified. The crude five year survival after elective surgery, including deaths within 30 days of surgery, was 79% for both men and women. When compared with a matched population the five year relative survival after elective surgery was 94.9% (95% confidence interval 89.9% to 99.9%) for men but only 88.0% (76.3% to 99.7%) for women. The five year relative survival of those aged 80 years and over was good: 116.6% (89.1% to 144.0%) compared with 92.4% (87.7% to 97.0%) for those under 80 years of age (men and women combined). Cardiovascular disease caused 57.8% of the 341 deaths after 30 days. CONCLUSION: In a condition such as abdominal aortic aneurysm, which occurs in elderly patients, relative survival is more clinically meaningful than crude survival. The five year relative survival in cases of elective and ruptured abdominal aortic aneurysm was better in men than in women. This is probably because of greater comorbidity in women with abdominal aortic aneurysm and this deserves more attention in the future. The long term survival outcome in octogenarians supports surgery in selected cases. PMID- 9748178 TI - Effect of sex of fetus on asthma during pregnancy: blind prospective study. PMID- 9748179 TI - A dangerous custom at funerals PMID- 9748180 TI - Surgical epidemic PMID- 9748181 TI - World health organisation PMID- 9748182 TI - Having your research ideas stolen PMID- 9748184 TI - Fashion and medicine PMID- 9748183 TI - Attributes of clinical guidelines that influence use of guidelines in general practice: observational study. AB - OBJECTIVE: To determine which attributes of clinical practice guidelines influence the use of guidelines in decision making in clinical practice. DESIGN: Observational study relating the use of 47 different recommendations from 10 national clinical guidelines to 12 different attributes of clinical guidelines for example, evidence based, controversial, concrete. SETTING: General practice in the Netherlands. SUBJECTS: 61 general practitioners who made 12 880 decisions in their contacts with patients. MAIN OUTCOME MEASURES: Compliance of decisions with clinical guidelines according to the attribute of the guideline. RESULTS: Recommendations were followed in, on average, 61% (7915/12 880) of the decisions. Controversial recommendations were followed in 35% (886/2497) of decisions and non-controversial recommendations in 68% (7029/10 383) of decisions. Vague and non-specific recommendations were followed in 36% (826/2280) of decisions and clear recommendations in 67% (7089/10 600) of decisions. Recommendations that demanded a change in existing practice routines were followed in 44% (1278/2912) of decisions and those that did not in 67% (6637/9968) of decisions. Evidence based recommendations were used more than recommendations for practice that were not based on research evidence (71% (2745/3841) v 57% (5170/9039)). CONCLUSIONS: People and organisations setting evidence based clinical practice guidelines should take into account some of the other important attributes of effective recommendations for clinical practice. PMID- 9748185 TI - Guidelines in general practice: the new Tower of Babel? PMID- 9748186 TI - Legibility of doctors' handwriting: quantitative comparative study. PMID- 9748187 TI - Netlines PMID- 9748188 TI - Male pattern androgenetic alopecia. PMID- 9748189 TI - Treatment resistant epilepsy or convulsive syncope? PMID- 9748191 TI - Tea for two and two for tea PMID- 9748190 TI - Oxygen therapy in chronic lung disease. PMID- 9748193 TI - Action on clinical audit: progress report 2. PMID- 9748192 TI - When placebo controlled trials are essential and equivalence trials are inadequate. PMID- 9748194 TI - Human albumin administration in critically ill patients. Evidence needs to be shown in paediatrics. PMID- 9748195 TI - Effects of the Heartbeat Wales programme. Effects of government policies on health behaviour must be studied. PMID- 9748196 TI - Antiretroviral combination therapy and HIV infection. Such treatment improved CD4 counts in Scottish patients. PMID- 9748198 TI - Mainuddin ahmed PMID- 9748197 TI - Technical ability to treat male factor infertility must not overtake academic knowledge. PMID- 9748199 TI - Question marks remain over PCGs PMID- 9748200 TI - Letter from america PMID- 9748201 TI - GPs and junkies PMID- 9748202 TI - Selling cheese-a beginner's guide PMID- 9748203 TI - Data, data, data. Give Me peace and knowledge PMID- 9748204 TI - No link exists between exposure to benzodiazepines in pregnancy and cleft palate PMID- 9748205 TI - Beer and spirits, but not wine, raise the risk of upper digestive tract cancer PMID- 9748207 TI - Men and octogenarians do well after surgery for abdominal aortic aneurysms PMID- 9748206 TI - Women later diagnosed with hyperparathyroidism take more sick leave PMID- 9748208 TI - Effective guidelines take implementation into account PMID- 9748209 TI - Asthma gets worse in women pregnant with girls PMID- 9748210 TI - Enzyme catalytic power minireview series. PMID- 9748211 TI - The low barrier hydrogen bond in enzymatic catalysis. PMID- 9748212 TI - Control of cell cycle progression by fibronectin matrix architecture. AB - Developmental patterning and differentiation, maintenance of parenchymal cell function, and the size, shape, and invasiveness of tumors are all orchestrated by cell interactions with the extracellular matrix. Here we show that the fibrillar structure of fibronectin (FN) matrix encodes essential regulatory cues and controls cell proliferation and signaling through changes in matrix architecture. A matrix assembled from native FN stimulated cell growth. In contrast, a mutant FN (FNDeltaIII1-7) that contains all known cell binding motifs but forms a structurally distinct matrix inhibited progression from G0/G1 into S phase. Furthermore, FNDeltaIII1-7 suppressed the stimulatory capacity of native FN and induced different levels of tyrosine phosphorylation of pp125(FAK). The differential effects on cell growth were ablated by blocking formation of matrix fibrils. Thus, modification of matrix architecture provides a novel approach to control cell proliferation. PMID- 9748213 TI - Polyunsaturated fatty acids decrease expression of promoters with sterol regulatory elements by decreasing levels of mature sterol regulatory element binding protein. AB - Membrane physiology, plasma lipid levels, and intracellular sterol homeostasis are regulated by both fatty acids and cholesterol. Sterols regulate gene expression of key enzymes of cholesterol and fatty acid metabolism through proteolysis of the sterol regulatory element-binding protein (SREBP), which binds to sterol regulatory elements (SRE) contained in promoters of these genes. We investigated the effect of fatty acids on SRE-dependent gene expression and SREBP. Consistent results were obtained in three different cell lines (HepG2, Chinese hamster ovary, and CV-1) transfected with SRE-containing promoters linked to the luciferase expression vector. We show that micromolar concentrations of oleate and other polyunsaturated fatty acids (C18:2-C22:6) dose-dependently (0.075-0.6 mmol) decreased transcription of SRE-regulated genes by 20-75%. Few or no effects were seen with saturated free fatty acids. Fatty acid effects on SRE dependent gene expression were independent and additive to those of exogenous sterols. Oleate decreased levels of the mature sterol regulatory element-binding proteins SREBP-1 and -2 and HMG-CoA synthase mRNA. Oleate had no effect in sterol regulation defective Chinese hamster ovary cells or in cells transfected with mutant SRE-containing promoters. We hypothesize that unsaturated fatty acids increase intracellular regulatory pools of cholesterol and thus affect mature SREBP levels and expression of SRE-dependent genes. PMID- 9748214 TI - A human RNA polymerase II transcription termination factor is a SWI2/SNF2 family member. AB - We obtained protein sequence information from Drosophila factor 2, an ATP dependent RNA polymerase II transcription termination factor, and discovered that it was identical to a SWI2/SNF2 family member called lodestar. Portions of putative human and Caenorhabditis elegans homologues were found in the sequence data bases and a complete cDNA for the human factor was generated using polymerase chain reaction techniques. Recombinant human factor 2 was produced in a baculovirus expression system, purified, and characterized. Similar to the authentic Drosophila factor, the human factor displayed a strong double-stranded DNA-dependent ATPase activity that was inhibited by single-stranded DNA and exhibited RNA polymerase II termination activity. Both factors were able to work on elongation complexes from either species. We discuss the mechanism of termination by factor 2 and the implications for the role of factor 2 in cellular activities. PMID- 9748215 TI - Reversibility of scrapie inactivation is enhanced by copper. AB - The only known difference between the cellular (PrPC) and scrapie-specific (PrPSc) isoforms of the prion protein is conformational. Because disruption of PrPSc structure decreases scrapie infectivity, restoration of the disease specific conformation should restore infectivity. In this study, disruption of PrPSc (as monitored by the loss of proteinase K resistance) by guanidine hydrochloride (GdnHCl) resulted in decreased infectivity. Upon dilution of the GdnHCl, protease resistance of PrP was restored and infectivity was regained. The addition of copper facilitated restoration of both infectivity and protease resistance of PrP in a subset of samples that did not renature by the simple dilution of the GdnHCl. These data demonstrate that loss of scrapie infectivity can be a reversible process and that copper can enhance this restoration of proteinase K resistance and infectivity. PMID- 9748216 TI - Phosphorylation of the cytoplasmic tail of syndecan-4 regulates activation of protein kinase Calpha. AB - Syndecans are transmembrane proteoglycans capable of carrying both heparan and chondroitin sulfate chains. The cytoplasmic tail of syndecan-4 was recently reported to undergo in vivo phosphorylation on Ser183 in the membrane-proximal part of the tail (Horowitz, A., and Simons, M. (1998) J. Biol. Chem. 273, 10914 10918). However, the functional consequences of this event remain unknown. The cytoplasmic tail of syndecan-4 is known to undergo multimerization and to activate protein kinase Calpha (PKCalpha), with both events depending on the presence of the commonly occurring phospholipid phosphatidylinositol 4,5 bisphosphate (PIP2). In the present investigation we found that phosphorylation of Ser183 produced a 10-fold reduction in the ability of syndecan-4 to activate PKCalpha, without affecting its ability to bind the PKC. Because Ser183 is adjacent to positively charged lysine groups that resemble PIP2-binding regions in several other proteins, phosphorylation of this serine may affect the binding affinity of the syndecan-4 cytoplasmic tail to PIP2. We found that the Ser183 phosphorylated cytoplasmic tail of syndecan-4 has indeed a significantly lower affinity to PIP2 compared with the nonphosphorylated tail. Furthermore, Ser183 phosphorylation abolished PIP2-dependent oligomerization of syndecan-4 cytoplasmic tails. We conclude that Ser183 phosphorylation regulates syndecan-4 dependent activation of PKCalpha by reducing the affinity to PIP2 and inhibiting the oligomerization of syndecan-4 cytoplasmic tails. These results further support the role of syndecan-4 in signal transduction in endothelial cells. PMID- 9748217 TI - The chaperone BiP/GRP78 binds to amyloid precursor protein and decreases Abeta40 and Abeta42 secretion. AB - Recent studies of cellular amyloid precursor protein (APP) metabolism demonstrate a beta-/gamma-secretase pathway resident to the endoplasmic reticulum (ER)/Golgi resulting in intracellular generation of soluble APP (APPsbeta) and Abeta42 peptide. Thus, these intracellular compartments may be key sites of amyloidogenic APP metabolism and Alzheimer's disease pathogenesis. We hypothesized that the ER chaperone immunoglobulin binding protein (BiP/GRP78) binds to and facilitates correct folding of nascent APP. Metabolic labeling and immunoprecipitation of transiently transfected human embryonic kidney 293 cells demonstrated co precipitation of APP with GRP78, revealing their transient interaction in the ER. Maturation of cellular APP was impaired by this interaction. Furthermore, the levels of APPs, Abeta40, and Abeta42 recovered in conditioned medium were lower compared with cells transfected with APP alone. Co-expression with APP of GRP78 T37G, an ATPase mutant, almost completely blocked cellular APP maturation as well as recovery of APPs, Abeta40, and Abeta42 in conditioned medium. The inhibitory effects of GRP78 and GRP78 T37G on Abeta40 and Abeta42 secretion were magnified by co-expression with the Swedish mutation of APP (K670N/M671L). Collectively, these data suggest a transient and direct interaction of GRP78 with APP in the ER that modulates intracellular APP maturation and processing and may facilitate its correct folding. PMID- 9748218 TI - Heparan sulfate/heparin N-deacetylase/N-sulfotransferase. The N-sulfotransferase activity domain is at the carboxyl half of the holoenzyme. AB - Glycosaminoglycan N-acetylglucosaminyl N-deacetylases/N-sulfotransferases are structurally related enzymes that play an important role in the biosynthesis of heparan sulfate and heparin. They are dual catalytic, single membrane-spanning polypeptides of approximately 850-880 amino acids that catalyze the N deacetylation of N-acetylglucosamine of glycosaminoglycans followed by N sulfation of the same sugar. On the basis of homologies of these proteins with other N-acetylglucosaminyl N-deacetylases involved in the biosynthesis of chitin and putative deacetylases from bacteria, we have constructed two soluble chimeras between protein A and the amino- and carboxyl-terminal halves of the above mastocytoma holoenzyme. The carboxyl-terminal chimera half (amino acids 479-880) was able to catalyze the N-sulfation of glucosamine of heparan sulfate with a similar affinity for its two substrates, adenosine 3'-phosphate 5'-phosphosulfate and heparan sulfate, as the holoenzyme. However, the reaction only occurred at 30 degreesC and not at 37 degreesC, both temperatures at which the holoenzyme was active. The Vmax of the chimera was 10-20-fold slower than that of the holoenzyme. Soluble chimeras between protein A and amino acids 43-521 and 43-680 of the holoenzyme were unable to catalyze the N-deacetylation of the bacterial N acetylglucosaminyl-glucuronic acid polymer K5 under conditions where the holoenzyme was active. The recent appearance in genome data banks of homologs to the N-sulfotransferase domain and now the direct demonstration that this domain catalyzes this reaction raises the possibility that both N-deacetylation and N sulfation activities of the holoenzyme might have emerged as gene fusions during evolution. PMID- 9748219 TI - Posttranslational formation of formylglycine in prokaryotic sulfatases by modification of either cysteine or serine. AB - Eukaryotic sulfatases carry an alpha-formylglycine residue that is essential for activity and is located within the catalytic site. This formylglycine is generated by posttranslational modification of a conserved cysteine residue. The arylsulfatase gene of Pseudomonas aeruginosa also encodes a cysteine at the critical position. This protein could be expressed in active form in a sulfatase deficient strain of P. aeruginosa, thereby restoring growth on aromatic sulfates as sole sulfur source, and in Escherichia coli. Analysis of the mature protein expressed in E. coli revealed the presence of formylglycine at the expected position, showing that the cysteine is also converted to formylglycine in a prokaryotic sulfatase. Substituting the relevant cysteine by a serine codon in the P. aeruginosa gene led to expression of inactive sulfatase protein, lacking the formylglycine. The machinery catalyzing the modification of the Pseudomonas sulfatase in E. coli therefore resembles the eukaryotic machinery, accepting cysteine but not serine as a modification substrate. By contrast, in the arylsulfatase of Klebsiella pneumoniae a formylglycine is found generated by modification of a serine residue. The expression of both the Klebsiella and the Pseudomonas sulfatases as active enzymes in E. coli suggests that two modification systems are present, or that a common modification system is modulated by a cofactor. PMID- 9748220 TI - The subsites structure of bovine pancreatic ribonuclease A accounts for the abnormal kinetic behavior with cytidine 2',3'-cyclic phosphate. AB - The kinetics of the hydrolysis of cytidine 2',3'-cyclic phosphate (C>p) to 3'-CMP by ribonuclease A are multiphasic at high substrate concentrations. We have investigated these kinetics by determining 3'-CMP formation both spectrophotometrically and by a highly specific and quantitative chemical sampling method. With the use of RNase A derivatives that lack a functional p2 binding subsite, evidence is presented that the abnormal kinetics with the native enzyme are caused by the sequential binding of the substrate to the several subsites that make up the active site of ribonuclease. The evidence is based on the following points. 1) Some of the unusual features found with native RNase A and C>p as substrate disappear when the derivatives lacking a functional p2 binding subsite are used. 2) The kcat/Km values with oligocytidylic acids of increasing lengths (ending in C>p) show a behavior that parallels the specific velocity with C>p at high concentrations: increase in going from the monomer to the trimer, a decrease from tetramer to hexamer, and then an increase in going to poly(C). 3) Adenosine increases the kcat obtained with a fixed concentration of C>p as substrate. 4) High concentrations of C>p protect the enzyme from digestion with subtilisin, which results in a more compact molecule, implying large substrate concentration-induced conformational changes. The data for the hydrolysis of C>p by RNase A can be fitted to a fifth order polynomial that has been derived from a kinetic scheme based on the sequential binding of several monomeric substrate molecules. PMID- 9748221 TI - Activation of proliferator-activated receptors alpha and gamma induces apoptosis of human monocyte-derived macrophages. AB - Peroxisome proliferator-activated receptors (PPARs) have been implicated in metabolic diseases, such as obesity, diabetes, and atherosclerosis, due to their activity in liver and adipose tissue on genes involved in lipid and glucose homeostasis. Here, we show that the PPARalpha and PPARgamma forms are expressed in differentiated human monocyte-derived macrophages, which participate in inflammation control and atherosclerotic plaque formation. Whereas PPARalpha is already present in undifferentiated monocytes, PPARgamma expression is induced upon differentiation into macrophages. Immunocytochemistry analysis demonstrates that PPARalpha resides constitutively in the cytoplasm, whereas PPARgamma is predominantly nuclear localized. Transient transfection experiments indicate that PPARalpha and PPARgamma are transcriptionally active after ligand stimulation. Ligand activation of PPARgamma, but not of PPARalpha, results in apoptosis induction of unactivated differentiated macrophages as measured by the TUNEL assay and the appearance of the active proteolytic subunits of the cell death protease caspase-3. However, both PPARalpha and PPARgamma ligands induce apoptosis of macrophages activated with tumor necrosis factor alpha/interferon gamma. Finally, PPARgamma inhibits the transcriptional activity of the NFkappaB p65/RelA subunit, suggesting that PPAR activators induce macrophage apoptosis by negatively interfering with the anti-apoptotic NFkappaB signaling pathway. These data demonstrate a novel function of PPAR in human macrophages with likely consequences in inflammation and atherosclerosis. PMID- 9748222 TI - Mediation of cyclic AMP signaling by the first intracellular loop of the gonadotropin-releasing hormone receptor. AB - The gonadotropin-releasing hormone (GnRH) receptor, which is a unique G protein coupled receptor without a C-terminal cytoplasmic domain, activates both inositol phosphate (InsP) and cAMP signaling responses. The function of the highly basic first intracellular (1i) loop of the GnRH receptor in signal transduction was evaluated by mutating selected residues located in its N and C termini. Replacements of Leu58, Lys59, Gln61, and Lys62 at the N terminus, and Leu73, Ser74, and Leu80 at the C terminus, caused no change in binding affinity. The agonist-induced InsP and cAMP responses of the Q61E and K59Q,K62Q receptors were also unaffected, but the L58A receptor showed a normal InsP response and an 80% decrease in cAMP production. At the C terminus, the InsP response of the L73R receptor was normal, but cAMP production was reduced by 80%. The EC50 for GnRH induced InsP responses of the S74E and L80A receptors was increased by about one order of magnitude, and the cAMP responses were essentially abolished. These findings indicate that cAMP signaling from the GnRH receptor is dependent on specific residues in the 1i loop that are not essential for activation of the phosphoinositide signaling pathway. PMID- 9748223 TI - Import into mitochondria, folding and retrograde movement of fumarase in yeast. AB - A single translation product of the FUM1 gene encoding fumarase is distributed between the cytosol and mitochondria of Saccharomyces cerevisiae. All fumarase translation products are targeted and processed in mitochondria before distribution. Here we show that targeting of fumarase is coupled to translation and initially involves insertion of the protein across the mitochondrial membranes and processing by the matrix protease. Rapid folding of fumarase may determine its requirement for coupling of its translocation with translation and unique route of distribution. The amino termini of most fumarase molecules are translocated across the mitochondrial membranes and processed. Unlike the in vivo situation where these molecules are released into the cytosol, in vitro they remain externally attached to the mitochondria, thereby positioned for release from the organelle. Our model suggests that fumarase displays a unique mechanism of targeting and distribution, which occurs cotranslationally and involves folding and retrograde movement of the processed protein back through the translocation pore. PMID- 9748224 TI - Phenylalanine hydroxylase from Chromobacterium violaceum. Uncoupled oxidation of tetrahydropterin and the role of iron in hyroxylation. AB - A gene encoding phenylalanine hydroxylase has been cloned from Chromobacterium violaceum and expressed in Escherichia coli. The purified phenylalanine hydroxylase contains copper, which does not support enzymatic activity. Upon removal of copper by dithiothreitol (DTT), the enzyme contains substoichiometric amounts of calcium and zinc but little or no redox-active metal ions. The copper depleted hydroxylase catalyzes the phenylalanine-dependent oxidation of 6, 7 dimethyltetrahydropterin (DMPH4) by O2 in a reaction in which phenylalanine is not hydroxylated and does not appear to undergo a chemical change, and hydrogen peroxide is produced. Analogs of phenylalanine also activate the oxidation of DMPH4. Both the copper-phenylalanine hydroxylase and the copper-depleted hydroxylase catalyze the hydroxylation of phenylalanine in the presence of DTT and FeSO4 in a reaction in which hydrogen peroxide is not produced. The apparent values of Km for Fe2+ and DTT are 0.28 microM and 1.1 mM, respectively, at 1.0 mM phenylalanine, 120 microM DMPH4 and pH 7. 4 and 23 degreesC. The apparent value of kcat is 14.3 s-1 under these conditions. Glutathione, mercaptoethanol, and dihydrolipoate support the hydroxylation of phenylalanine essentially as well as DTT. Incubation of copper-depleted hydroxylase with FeSO4, phenylalanine, and DTT followed by gel permeation chromatography leads to an iron-hydroxylase containing approximately 1 molecule of iron per molecule of enzyme. The iron-hydroxylase displays an optical absorption band extending from 300 to 600 nm, and it catalyzes the hydroxylation of phenylalanine at the same maximum rate as the iron activated hydroxylase but does not require added Fe2+. We conclude that iron participates in the hydroxylation of phenylalanine. Iron is not required for the oxidation of DMPH4, although it may exert a modest acceleration effect. A hypothetical mechanism is presented wherein the reaction of iron with the putative 4a-hydroperoxy-DMPH4 leads to 4a-hydroxy-DMPH4 and a high valent iron oxy species. The iron-oxy species is postulated to react with phenylalanine in the hydroxylation process. PMID- 9748225 TI - Synergy between actin depolymerizing factor/cofilin and profilin in increasing actin filament turnover. AB - The mechanism of control of the steady state of actin assembly by actin depolymerizing factor (ADF)/cofilin and profilin has been investigated. Using Tbeta4 as an indicator of the concentration of ATP-G-actin, we show that ADF increases the concentration of ATP-G-actin at steady state. The measured higher concentration of ATP-G-actin is quantitatively consistent with the increase in treadmilling, caused by the large increase in the rate of depolymerization from the pointed ends induced by ADF (Carlier, M.-F. , Laurent, V., Santolini, J., Didry, D., Melki, R., Xia, G.-X., Hong, Y., Chua, N.-H., and Pantaloni, D. (1997) J. Cell Biol. 136, 1307-1322). Experiments demonstrate that profilin synergizes with ADF to further enhance the turnover of actin filaments up to a value 125 fold higher than in pure F-actin solutions. Profilin and ADF act at the two ends of filaments in a complementary fashion to increase the processivity of treadmilling. Using the capping protein CapZ, we show that ADF increases the number of filaments at steady state by 1. 3-fold, which cannot account for the 25 fold increase in turnover rate. Computer modeling of the combined actions of ADF and profilin on the dynamics of actin filaments using experimentally determined rate constants generates a distribution of the different actin species at steady state, which is in quantitative agreement with the data. PMID- 9748226 TI - Amino acids 430-570 in apolipoprotein B are critical for its binding to microsomal triglyceride transfer protein. AB - Several studies have demonstrated protein-protein interactions between microsomal triglyceride transfer protein (MTP) and apolipoprotein B (apoB). However, the binding sites involved in these interactions have not been elucidated. To identify an MTP binding site in apoB, we have expressed several apoB sequences as fusion proteins with the eight-amino acid FLAG peptide. The chimeras were transiently expressed in COS cells, and conditioned media were used to study the binding of these sequences to either immobilized or soluble MTP. A polypeptide containing amino acids 270-570 (B:270-570), but not 1-300, bound to MTP. AGI-S17, an antagonist of apoB-MTP binding, inhibited the binding of B:270-570 to MTP but not to M2, a monoclonal antibody that recognizes the FLAG peptide. These data indicated that B:270-570 contains an MTP binding site. Next, sequences within 270 570 were subjected to C-terminal truncations at natural proline residues. B:270 509 bound less efficiently than B:270-570, whereas, B:270-430 and other shorter chimeras did not bind to MTP. Furthermore, truncations at amino acids 502 and 509 decreased MTP binding by 73 and 42%, respectively. These data indicate that B:430 570 in the alpha1-globular domain of apoB plays a crucial role in MTP binding and presumably in the initiation and maturation of apoB-containing lipoproteins. PMID- 9748227 TI - Identification of sorting determinants in the C-terminal cytoplasmic tails of the gamma-aminobutyric acid transporters GAT-2 and GAT-3. AB - In order to perform their physiologic functions, polarized epithelial cells must target ion transport proteins to the appropriate domains of their plasma membranes. Molecular signals responsible for polarized sorting have been identified for several membrane proteins which span the bilayer once. Most ion transport proteins are polytopic, however, and little is known of the signals responsible for the targeting of this class of polypeptides. Members of the gamma aminobutyric acid (GABA) transporter family are polytopic membrane proteins found endogenously in both epithelial cells and neurons. We have identified narrowly defined sequences which are required for the proper accumulation of two members of this transporter family in Madin-Darby canine kidney cells. The highly homologous GABA transporter isoforms, GAT-2 and GAT-3, localize to the basolateral and apical surfaces, respectively, when expressed stably in Madin Darby canine kidney cells. We have generated deletion constructs and chimeric transporters composed of complimentary portions of GAT-2 and GAT-3. We find that information which directs their differential sorting is present in the C-terminal cytoplasmic tails of these two polypeptides. A sequence of 22 amino acids at the C terminus of GAT-2 is required for the transporter's basolateral distribution and is capable of directing GAT-3 to the basolateral surface when appended to the C terminus of this normally apical polypeptide. The deletion of 32 amino acids from the C terminus of GAT-3 causes this transporter to become mislocalized to both surfaces. Moreover, removal of the final three amino acids of GAT-3 (THF) similarly disrupts its apical sorting. The GAT-3 C-terminal sequence resembles motifs which interact with PDZ domains, raising the possibility that the steady state distribution of GAT-3 at the apical plasmalemmal surface requires a protein protein interaction mediated by its extreme C-terminal cytoplasmic tail. These data provide the first characterization of a protein-based signal required for the apical distribution of a membrane protein. PMID- 9748228 TI - Specific activation of Smad1 signaling pathways by the BMP7 type I receptor, ALK2. AB - BMP7 and activin are members of the transforming growth factor beta superfamily. Here we characterize endogenous activin and BMP7 signaling pathways in P19 embryonic carcinoma cells. We show that BMP7 and activin bind to the same type II receptors, ActRII and IIB, but recruit distinct type I receptors into heteromeric receptor complexes. The major BMP7 type I receptor observed was ALK2, while activin bound exclusively to ALK4 (ActRIB). BMP7 and activin elicited distinct biological responses and activated different Smad pathways. BMP7 stimulated phosphorylation of endogenous Smad1 and 5, formation of complexes with Smad4 and induced the promoter for the homeobox gene, Tlx2. In contrast, activin induced phosphorylation of Smad2, association with Smad4, and induction of the activin response element from the Xenopus Mix.2 gene. Biochemical analysis revealed that constitutively active ALK2 associated with and phosphorylated Smad1 on the COOH terminal SSXS motif, and also regulated Smad5 and Smad8 phosphorylation. Activated ALK2 also induced the Tlx2 promoter in the absence of BMP7. Furthermore, we show that ALK1 (TSRI), an orphan receptor that is closely related to ALK2 also mediates Smad1 signaling. Thus, ALK1 and ALK2 induce Smad1-dependent pathways and ALK2 functions to mediate BMP7 but not activin signaling. PMID- 9748229 TI - Contribution of proteasomal beta-subunits to the cleavage of peptide substrates analyzed with yeast mutants. AB - Proteasomes generate peptides that can be presented by major histocompatibility complex (MHC) class I molecules in vertebrate cells. Using yeast 20 S proteasomes carrying different inactivated beta-subunits, we investigated the specificities and contributions of the different beta-subunits to the degradation of polypeptide substrates containing MHC class I ligands and addressed the question of additional proteolytically active sites apart from the active beta-subunits. We found a clear correlation between the contribution of the different subunits to the cleavage of fluorogenic and long peptide substrates, with beta5/Pre2 cleaving after hydrophobic, beta2/Pup1 after basic, and beta1/Pre3 after acidic residues, but with the exception that beta2/Pup1 and beta1/Pre3 can also cleave after some hydrophobic residues. All proteolytic activities including the "branched chain amino acid-preferring" component are associated with beta5/Pre2, beta1/Pre3, or beta2/Pup1, arguing against additional proteolytic sites. Because of the high homology between yeast and mammalian 20 S proteasomes in sequence and subunit topology and the conservation of cleavage specificity between mammalian and yeast proteasomes, our results can be expected to also describe most of the proteolytic activity of mammalian 20 S proteasomes leading to the generation of MHC class I ligands. PMID- 9748230 TI - Imidazole binding to Rhodobacter capsulatus cytochrome c2. Effect of site directed mutants on ligand binding. AB - Although ligand binding in c-type cytochromes is not directly related to their physiological function, it has the potential to provide valuable information on protein stability and dynamics, particularly in the region of the methionine sixth heme ligand and the nearby peptide chain that has been implicated in electron transfer. Thus, we have measured the equilibrium and kinetics of binding of imidazole to eight mutants of Rhodobacter capsulatus cytochrome c2 that differ in overall protein stability. We found that imidazole binding affinity varies 70 fold, but does not correlate with overall protein stability. Instead, each mutant exerts an effect at the local level, with the largest change due to mutant G95E (glycine substituted by glutamate), which shows 30-fold stronger binding as compared with the wild-type protein. The kinetics of imidazole binding are monophasic and reach saturation at high ligand concentrations for all the mutants and wild-type protein, which is attributed to a rate-limiting conformational change leading to breakage of the iron-methionine bond and providing a binding site for imidazole. The mutants show as much as an 18-fold variation in the first order rate constant for the conformational change, with the largest effect found with mutant G95E. The kinetics also show a lack of correlation with overall protein stability, but are consistent with localized effects on the dynamics of hinge region 88-102 of the protein, which changes conformation to permit ligand binding. These results are consistent with R. capsulatus cytochrome c2 stabilizing the complex through hydrogen bonding to the imidazole. The larger effects of mutant G95E on equilibrium and kinetics are likely to be due to its location within the hinge region adjacent to heme ligand methionine 96, which is displaced by imidazole. PMID- 9748231 TI - Regulation of DNA-dependent protein kinase by the Lyn tyrosine kinase. AB - The Src-like protein-tyrosine kinase Lyn is activated by ionizing radiation and certain other DNA-damaging agents, whereas the DNA-dependent protein kinase (DNA PK), consisting of the catalytic subunits (DNA-PKcs) and Ku DNA-binding components, requires DNA double-stranded breaks for activation. Here we demonstrate that Lyn associates constitutively with DNA-PKcs. The SH3 domain of Lyn interacts directly with DNA-PKcs near a leucine zipper homology domain. We also show that Lyn phosphorylates DNA-PKcs but not Ku in vitro. The interaction between Lyn and DNA-PKcs inhibits DNA-PKcs activity and the ability of DNA-PKcs to form a complex with Ku/DNA. These results support the hypothesis that there are functional interactions between Lyn and DNA-PKcs in the response to DNA damage. PMID- 9748232 TI - Lipid binding ridge on loops 2 and 3 of the C2A domain of synaptotagmin I as revealed by NMR spectroscopy. AB - The C2A domain of synaptotagmin I, which binds Ca2+ and anionic phospholipids, serves as a Ca2+ sensor during excitation-secretion coupling. We have used multidimensional NMR to locate the region of C2A from rat synaptotagmin I that interacts, in the presence of Ca2+, with phosphatidylserine. Untagged, recombinant C2A was double-labeled with 13C and 15N, and triple-resonance NMR data were collected from C2A samples containing either Ca2+ alone or Ca2+ plus 6:0 phosphatidylserine. Phospholipid binding led to changes in chemical shifts of backbone atoms in residues Arg233 and Phe234 of loop 3 (a loop that also binds Ca2+) and His198, Val205, and Phe206 of loop 2. These residues lie along a straight line on a surface ridge of the C2A domain. The only other residue that exhibited appreciable chemical shift changes upon adding lipid was His254; however, because His254 is located on the other side of the molecule from the phospholipid docking site defined by the other residues, its shifts may result from nonspecific interactions. The results show that the "docking ridge" responsible for Ca2+-dependent membrane association is localized on the opposite side of the C2A domain from the transmembrane and C2B domains of synaptotagmin. PMID- 9748233 TI - Identification of amino acid residues that form part of the ligand-binding pocket of integrin alpha5 beta1. AB - Arg-Arg-Glu-Thr-Ala-Trp-Ala (RRETAWA) is a novel ligand peptide for integrin alpha5 beta1, which blocks alpha5 beta1-mediated cell adhesion to fibronectin (Koivunen, E., Wang, B., and Ruoslahti, E. (1994) J. Cell Biol. 124, 373-380). Here we have localized the binding site for RRETAWA on alpha5 beta1 using inhibitory monoclonal antibodies (mAbs) and site-directed mutagenesis. A cyclic peptide containing this sequence (*CRRETAWAC*) had little effect on the binding of most anti-alpha5 and anti-beta1 mAbs to alpha5 beta1 but completely blocked binding of the anti-alpha5 mAb 16 in a directly competitive manner. Hence, the binding site of RRETAWA appears to closely overlap with the epitope of mAb 16. *CRRETAWAC* also acted as a direct competitive inhibitor of the binding of Arg Gly-Asp (RGD)-containing fibronectin fragments to alpha5 beta1, suggesting that the binding site for RRETAWA is also closely overlapping with that for RGD. However, differences between the binding sites of RRETAWA and RGD were apparent in that (i) RGD peptides allosterically inhibited the binding of mAb 16 to alpha5 beta1, and (ii) several mAbs that perturbed binding of alpha5 beta1 to RGD had little effect on binding of alpha5 beta1 to RRETAWA. A double mutation in alpha5 (S156G/W157S) blocked the interaction of both RRETAWA and mAb 16 with alpha5 beta1 but had no effect on fibronectin binding or on the binding of other anti alpha5 mAbs. Ser156-Trp157 is located near the apex of a putative loop region on the upper surface of a predicted beta-propeller structure formed by the NH2 terminal repeats of alpha5. Our findings suggest that this sequence forms part of the ligand-binding pocket of alpha5 beta1. Furthermore, as Ser156-Trp157 is unique to the alpha5 subunit, it may be responsible for the specific recognition of RRETAWA by alpha5 beta1. PMID- 9748234 TI - Direct binding of p130(Cas) to the guanine nucleotide exchange factor C3G. AB - p130(Cas) (Cas; crk-associated substrate) belongs to a new family of docking molecules. It contains one Src homology (SH) 3 domain in its amino-terminal region followed by a region containing binding motifs for SH2 and SH3 domains. To gain further insight into Cas signaling we used the SH3 domain of Cas in a two hybrid screen to search a human placenta library for binding partners. The screen confirmed a previous finding of its binding to the focal adhesion kinase (FAK) but also identified C3G, a guanine nucleotide exchange factor. We found direct interaction between Cas and C3G in vitro and in vivo. A series of analysis with C3G deletion mutants revealed a proline-rich Cas-binding site (Ala0-Pro1-Pro2 Lys3-Pro4-Pro5-Leu6-Pro7) located NH2-terminal to the previously characterized Crk binding motifs in C3G. Mutagenesis studies showed that Pro1, Lys3, and Pro4 within the ligand-binding site are critical for high affinity interaction. These results, combined with sequence alignments of proline-rich binding elements from proteins known for Cas binding, define the consensus sequence XXPXKPX which is recognized by the CasSH3 domain. Cas shows structural characteristics of a docking molecule and may serve to bring C3G to specific compartments within the cell. PMID- 9748235 TI - The human chitotriosidase gene. Nature of inherited enzyme deficiency. AB - The human chitinase, named chitotriosidase, is a member of family 18 of glycosylhydrolases. Following the cloning of the chitotriosidase cDNA (Boot, R. G., Renkema, G. H., Strijland, A., van Zonneveld, A. J., and Aerts, J. M. F. G. (1995) J. Biol. Chem. 270, 26252-26256), the gene and mRNA have been investigated. The chitotriosidase gene is assigned to chromosome 1q31-q32. The gene consists of 12 exons and spans about 20 kilobases. The nature of the common deficiency in chitotriosidase activity is reported. A 24-base pair duplication in exon 10 results in activation of a cryptic 3' splice site, generating a mRNA with an in-frame deletion of 87 nucleotides. All chitotriosidase-deficient individuals tested were homozygous for the duplication. The observed carrier frequency of about 35% indicates that the duplication is the predominant cause of chitotriosidase deficiency. The presence of the duplication in individuals from various ethnic groups suggests that this mutation is relatively old. PMID- 9748236 TI - Action currents generate stepwise intracellular Ca2+ patterns in a neuroendocrine cell. AB - It is believed that specific patterns of changes in the cytosolic-free calcium concentration ([Ca2+]i) are used to control cellular processes such as gene transcription, cell proliferation, differentiation, and secretion. We recently showed that the Ca2+ oscillations in the neuroendocrine melanotrope cells of Xenopus laevis are built up by a number of discrete Ca2+ rises, the Ca2+ steps. The origin of the Ca2+ steps and their role in the generation of long-lasting Ca2+ patterns were unclear. By simultaneous, noninvasive measuring of melanotrope plasma membrane electrical activity and the [Ca2+]i, we show that numbers, amplitude, and frequency of Ca2+ steps are variable among individual oscillations and are determined by the firing pattern and shape of the action currents. The general Na+ channel blocker tetrodotoxin had no effect on either action currents or the [Ca2+]i. Under Na+-free conditions, a depolarizing pulse of 20 mM K+ induced repetitive action currents and stepwise increases in the [Ca2+]i. The Ca2+ channel blocker CoCl2 eliminated action currents and stepwise increases in the [Ca2+]i in both the absence and presence of high K+. We furthermore demonstrate that the speed of Ca2+ removal from the cytoplasm depends on the [Ca2+]i, also between Ca2+ steps during the rising phase of an oscillation. It is concluded that Ca2+ channels, and not Na+ channels, are essential for the generation of specific step patterns and, furthermore, that the frequency and shape of Ca2+ action currents in combination with the Ca2+ removal rate determine the oscillatory pattern. PMID- 9748237 TI - Cytochrome c heme lyase activity of yeast mitochondria. AB - A highly efficient in vitro system was established for measuring by high performance liquid chromatography the formation of holocytochrome c by yeast mitochondria. Holocytochrome c formation required reducing agents, of which dithiothreitol was the most effective. With biosynthetically made, pure Drosophila melanogaster apocytochrome c and Saccharomyces cerevisiae mitochondria, the activity of cytochrome c heme lyase amounted to about 800 fmol min-1 mg-1 mitochondrial protein. The kinetics were typical Michaelis-Menten (Km approximately 1 nM), as were those of mitoplasts with broken outer membranes (Km approximately 3 nM). As tested with mitoplasts, holocytochromes c from a range of species were found to be competitive inhibitors of heme lyase at physiological concentrations, providing a mechanism for controlling this concentration in vivo. Apocytochrome c associated with yeast mitochondria in two phases of Kd approximately 2 x 10(-10) and 10(-8) M, respectively, whereas mitoplasts had lost the high affinity binding. A site-directed mutant of apocytochrome c (lysines 5, 7, and 8 replaced by glutamine, glutamic acid, and asparagine) was found to be converted to holocytochrome c (Km approximately 3.3 nM; maximal activity unchanged), even though the mutations completely eliminated the high affinity binding. Thus, the high affinity binding of apocytochrome c to mitochondria is not directly related to holocytochrome c formation. PMID- 9748238 TI - Crystallographic and spectroscopic studies of native, aminoquinol, and monovalent cation-bound forms of methylamine dehydrogenase from Methylobacterium extorquens AM1. AB - Various monovalent cations influence the enzymatic activity and the spectroscopic properties of methylamine dehydrogenase (MADH). Here, we report the structure determination of this tryptophan tryptophylquinone-containing enzyme from Methylobacterium extorquens AM1 by high resolution x-ray crystallography (1.75 A). This first MADH crystal structure at low ionic strength is compared with the high resolution structure of the related MADH from Paracoccus denitrificans recently reported. We also describe the first structures (at 1.95 to 2.15 A resolution) of an MADH in the substrate-reduced form and in the presence of trimethylamine and of cesium, two competitive inhibitors. Polarized absorption microspectrophotometry was performed on single crystals under various redox, pH, and salt conditions. The results show that the enzyme is catalytically active in the crystal and that the cations cause the same spectral perturbations as are observed in solution. These studies lead us to propose a model for the entrance and binding of the substrate in the active site. PMID- 9748239 TI - The nuclear receptors peroxisome proliferator-activated receptor alpha and Rev erbalpha mediate the species-specific regulation of apolipoprotein A-I expression by fibrates. AB - Fibrates are widely used hypolipidemic drugs which activate the nuclear peroxisome proliferator-activated receptor (PPAR) alpha and thereby alter the transcription of genes controlling lipoprotein metabolism. Fibrates influence plasma high density lipoprotein and its major protein, apolipoprotein (apo) A-I, in an opposite manner in man (increase) versus rodents (decrease). In the present study we studied the molecular mechanisms of this species-specific regulation of apoA-I expression by fibrates. In primary rat and human hepatocytes fenofibric acid, respectively, decreased and increased apoA-I mRNA levels. The absence of induction of rat apoA-I gene expression by fibrates is due to 3 nucleotide differences between the rat and the human apoA-I promoter A site, rendering a positive PPAR-response element in the human apoA-I promoter nonfunctional in rats. In contrast, rat, but not human, apoA-I transcription is repressed by the nuclear receptor Rev-erbalpha, which binds to a negative response element adjacent to the TATA box of the rat apoA-I promoter. In rats fibrates increase liver Rev-erbalpha mRNA levels >10-fold. In conclusion, the opposite regulation of rat and human apoA-I gene expression by fibrates is linked to differences in cis-elements in their respective promoters leading to repression by Rev-erbalpha of rat apoA-I and activation by PPARalpha of human apoA-I. Finally, Rev-erbalpha is identified as a novel fibrate target gene, suggesting a role for this nuclear receptor in lipid and lipoprotein metabolism. PMID- 9748240 TI - Substrate recognition of tRNA (Guanosine-2'-)-methyltransferase from Thermus thermophilus HB27. AB - Transfer RNA (guanosine-2'-)-methyltransferase (Gm-methylase, EC 2.1. 1.32) from Thermus thermophilus HB27 is one of the tRNA ribose modification enzymes. The broad substrate specificity of Gm-methylase has so far been elucidated using various species of tRNAs from native sources, suggesting that the common structures in tRNAs are recognized by the enzyme. In this study, by using 28 yeast tRNAPhe variants obtained by transcription with T7 RNA polymerase, it was revealed that the nucleotide residues G18 and G19 and the D-stem structure are essentially required for Gm-methylase recognition, and that the key sequence for the substrate is pyrimidine (Py)17G18G19. The other conserved sequences were found not to be essential, but U8, G15, G26, G46, U54, U55, and C56 considerably affected the methylation efficiency. These residues are located within a limited space embedded in the L-shaped three-dimensional structure of tRNA. Therefore, disruption of the three-dimensional structure of the substrate tRNA is necessary for the catalytic center of Gm-methylase to be able to access the target site in the tRNA, suggesting that the interaction of Gm-methylase with tRNA consists of multiple steps. This postulation was confirmed by inhibition experiments using nonsubstrate tRNA variants which functioned as competitive inhibitors against usual substrate tRNAs. PMID- 9748241 TI - Negative regulation of Rho family GTPases Cdc42 and Rac2 by homodimer formation. AB - The Rho family GTPases are tightly regulated between the active GTP-bound state and the inactive GDP-bound state in a variety of signal transduction processes. Here the Rho family members Cdc42, Rac2, and RhoA were found to form reversible homodimers in both the GTP- and the GDP-bound states. The homophilic interaction of Cdc42 and Rac2, but not RhoA, in the GTP-bound state, caused a significant stimulation of the intrinsic GTPase activity, i.e. the activated form of Cdc42 and Rac2 acts as GTPase-activating proteins toward Cdc42-GTP or Rac2-GTP. The dimerization of the GTPases appeared to be mediated by the carboxyl-terminal polybasic domain, and the specific GTPase-activating effects of Cdc42 and Rac2 were also attributed to the structural determinant(s) in the same region of the molecules. Moreover, similar to the case of Cdc42 and Cdc42GAP interaction, Cdc42 GDP interacted with tetrafluoroaluminate and Cdc42-GTPgammaS (guanosine 5'-3-O (thio)triphosphate) to form a transition state complex of the GTPase-activating reaction in which the carboxyl-terminal determinant(s) of the GTPgammaS-bound Cdc42 plays a critical role. These results provide a rationale for the fast rate of intrinsic GTP hydrolysis by Cdc42 and Rac and suggest that dimerization may play a role in the negative regulation of specific Rho family GTPases mediated by the carboxyl-terminal polybasic domain. PMID- 9748242 TI - A ubiquinone-binding site regulates the mitochondrial permeability transition pore. AB - We have investigated the regulation of the mitochondrial permeability transition pore (PTP) by ubiquinone analogues. We found that the Ca2+-dependent PTP opening was inhibited by ubiquinone 0 and decylubiquinone, whereas all other tested quinones (ubiquinone 5, 1,4-benzoquinone, 2-methoxy-1,4-benzoquinone, 2,3 dimethoxy-1, 4-benzoquinone, and 2,3-dimethoxy-5,6-dimethyl-1,4-benzoquinone) were ineffective. Pore inhibition was observed irrespective of the method used to induce the permeability transition (addition of Pi or atractylate, membrane depolarization, or dithiol cross-linking). Inhibition of PTP opening by decylubiquinone was comparable with that exerted by cyclosporin A, whereas ubiquinone 0 was more potent. Ubiquinone 5, which did not inhibit the PTP per se, specifically counteracted the inhibitory effect of ubiquinone 0 or decylubiquinone but not that of cyclosporin A. These findings define a ubiquinone binding site directly involved in PTP regulation and indicate that different quinone structural features are required for binding and for stabilizing the pore in the closed conformation. At variance from all other quinones tested, decylubiquinone did not inhibit respiration. Our results define a new structural class of pore inhibitors and may open new perspectives for the pharmacological modulation of the PTP in vivo. PMID- 9748243 TI - Superoxide dependence of the toxicity of short chain sugars. AB - Erythrose inhibited the growth of a sodA sodB strain of Escherichia coli under aerobiosis; but did not inhibit anaerobic growth of the sodA sodB strain, or the aerobic growth of the superoxide dismutase (SOD)-competent parental strain. A SOD mimic protected the sodA sodB strain against the toxicity of erythrose as did the carbonyl-blocking reagents hydrazine and aminoguanidine. Three carbon sugars, such as glyceraldehyde and dihydroxy acetone, and the two carbon sugar glycolaldehyde, were similarly toxic in an O-2-dependent manner. An unidentified dialyzable component in E. coli extract augmented the oxidation of short chain sugars, and this was partially inhibitable by SOD. The toxicity of the short chain sugars appears to be because of an O-2-dependent oxidation to alpha, beta dicarbonyl compounds. In keeping with this view was the O-2-independent toxicity of methylglyoxal. PMID- 9748244 TI - The glucose transporter of Escherichia coli with circularly permuted domains is active in vivo and in vitro. AB - The bacterial phosphotransferase system (PTS) consists of two energy-coupling soluble proteins (enzyme I and HPr) and a large number of inner membrane transporters (enzymes II) that mediate concomitant phosphorylation and translocation of sugars and hexitols. The transporters consist of three functional units (IIA, IIB, IIC), which occur either as protein subunits or domains of a multidomain polypeptide. The membrane-spanning IIC domain contains the substrate binding site; IIA and IIB are phosphorylation domains that transfer phosphate from HPr to the transported sugar. The transporter complexes of the PTS are good examples for variation of design by modular assembly of domains and subunits. The domain order is IIC-IIB in the membrane subunit of the Escherichia coli glucose transporter (IICBGlc) and IIB-IIC in Salmonella typhimurium sucrose transporter (IIBCScr). The phosphorylation domain of IICBGlc was translocated from the carboxyl-terminal to the amino-terminal end of the IIC domain, and the activity of the circularly permuted form was optimized by variation of the length and the composition of the interdomain linker. IIBapCGlc with an alanine-proline rich interdomain linker has 70% of the control specific activity after purification and reconstitution into proteoliposomes. These results indicate that the amino-terminal end of IICBGlc must be on the cytoplasmic side of the inner membrane, that membrane insertion of the IIC domain is insensitive to the modification of its amino-terminal end, and that a domain swap as it could occur by a single DNA translocation event can rapidly lead to a functional protein. However, IIB could not be substituted for by glucokinase. Fusion proteins between the IIC domain and glucokinase do not transport and phosphorylate glucose in an ATP-dependent mechanism, although the IIC moiety displays transport activity upon complementation with soluble subclonal IIB, and the glucokinase moiety retains ATP-dependent nonvectorial kinase activity. This indicates that IIC and IIB are two cooperative units and not only sequentially acting upon a common substrate, and that translocation of glucose must be conformationally coupled to the phosphorylation/dephosphorylation cycle of IIB. PMID- 9748245 TI - An investigation of the metabolism of valine to isobutyl alcohol in Saccharomyces cerevisiae. AB - The metabolism of valine to isobutyl alcohol in yeast was examined by 13C nuclear magnetic resonance spectroscopy and combined gas chromatography-mass spectrometry. The product of valine transamination, alpha-ketoisovalerate, had four potential routes to isobutyl alcohol. The first, via branched-chain alpha ketoacid dehydrogenase to isobutyryl-CoA is not required for the synthesis of isobutyl alcohol because abolition of branched-chain alpha-ketoacid dehydrogenase activity in an lpd1 disruption mutant did not prevent the formation of isobutyl alcohol. The second route, via pyruvate decarboxylase, is the one that is used because elimination of pyruvate decarboxylase activity in a pdc1 pdc5 pdc6 triple mutant virtually abolished isobutyl alcohol production. A third potential route involved alpha-ketoisovalerate reductase, but this had no role in the formation of isobutyl alcohol from alpha-hydroxyisovalerate because cell homogenates could not convert alpha-hydroxyisovalerate to isobutyl alcohol. The final possibility, use of the pyruvate decarboxylase-like enzyme encoded by YDL080c, seemed to be irrelevant, because a strain with a disruption in this gene produced wild-type levels of isobutyl alcohol. Thus there are major differences in the catabolism of leucine and valine to their respective "fusel" alcohols. Whereas in the catabolism of leucine to isoamyl alcohol the major route is via the decarboxylase encoded by YDL080c, any single isozyme of pyruvate decarboxylase is sufficient for the formation of isobutyl alcohol from valine. Finally, analysis of the 13C labeled products revealed that the pathways of valine catabolism and leucine biosynthesis share a common pool of alpha-ketoisovalerate. PMID- 9748246 TI - Redox-controlled ligand exchange of the heme in the CO-sensing transcriptional activator CooA. AB - The transcriptional activator CooA from Rhodospirillum rubrum contains a b-type heme that acts as a CO sensor in vivo. CooA is the first example of a transcriptional regulator containing a heme as a prosthetic group and of a hemeprotein in which CO plays a physiological role. In this study, we constructed an in vivo reporter system to measure the transcriptional activator activity of CooA and prepared some CooA mutants in which a mutation was introduced at Cys, His, Met, Lys, or Tyr. Only the mutations of Cys75 and His77 affected the electronic absorption spectra of the heme in CooA. The electronic absorption spectra, EPR spectra, and the transcriptional activator activity of the wild-type and mutant CooA proteins indicate that 1) the thiolate derived from Cys75 is the axial ligand in the ferric heme, but it is not coordinated to the CO-bound ferrous heme; 2) Cys75 is protonated or displaced in the ferrous heme; and 3) His77 is the proximal ligand in the CO-bound ferrous heme and probably also in the ferrous heme, but it is not coordinated to the ferric heme. NMR spectra reveal that the conformational change around the heme, which will trigger the activation of CooA by CO, takes place upon the binding of CO to the heme. PMID- 9748247 TI - Extreme C terminus of G protein alpha-subunits contains a site that discriminates between Gi-coupled metabotropic glutamate receptors. AB - Metabotropic glutamate receptors (mGlu receptors), the Ca2+-sensing receptor, gamma-aminobutyric acid type B receptors, and one group of pheromone receptors constitute a unique family (also called family 3) of heptahelical receptors. This original family shares no sequence similarity with any other G protein-coupled receptors. The identification and comparison of the molecular determinants of receptor/G protein coupling within the different receptor families may help identify general rules involved in this protein/protein interaction. In order to detect possible contact sites important for coupling selectivity between family 3 receptors and the G protein alpha-subunits, we examined the coupling of the cyclase-inhibiting mGlu2 and mGlu4 receptors to chimeric alphaq-subunits bearing the 5 extreme C-terminal amino acid residues of either Galphai, Galphao, or Galphaz. Whereas mGlu4 receptor activated all three chimeric G proteins, mGlu2 receptor activated Galphaqi and Galphaqo but not Galphaqz. The mutation of isoleucine -4 of Galphaqz into cysteine was sufficient to recover coupling of the mutant G protein to mGlu2 receptor. Moreover, the mutation of cysteine -4 of Galphaqo into isoleucine was sufficient to suppress the coupling to mGlu2 receptor. Mutations at positions -5 and -1 had an effect on coupling efficiency, but not selectivity. Our results emphasize the importance of the residue -4 of the alpha-subunits in their specific interaction to heptahelical receptors by extending this finding on the third family of G protein-coupled receptors. PMID- 9748248 TI - The role of tyrosine phosphorylation of cortactin in the locomotion of endothelial cells. AB - Cortactin, a filamentous actin cross-linking protein and a substrate of Src protein tyrosine kinase, is phosphorylated at tyrosine residues upon stimulation by extracellular signals. We have previously demonstrated that the filamentous actin cross-linking activity of cortactin is attenuated by Src (Huang, C., Ni, Y., Gao, Y., Haudenschild, C. C., and Zhan, X. (1997) J. Biol. Chem. 272, 13911 13915). In vitro, tyrosine phosphorylation of cortactin occurs specifically within the region between the proline-rich sequence and the Src homology 3 domain. Among the nine tyrosine residues in this region, mutations at Tyr421, Tyr466, and Tyr482 significantly reduced Src-meditated tyrosine phosphorylation both in vitro and in vivo. Ectopic expression of wild-type cortactin in ECV304, a spontaneously transformed human umbilical endothelial cell line, resulted in an enhanced cell migration. In contrast, overexpression of a cortactin mutant deficient in tyrosine phosphorylation impaired the migration of endothelial cells. These findings reveal an intracellular signaling mechanism whereby the motility of endothelial cells is regulated by a Src-mediated tyrosine phosphorylation of cortactin. PMID- 9748249 TI - Tyrosine phosphorylation of focal adhesion kinase stimulated by hepatocyte growth factor leads to mitogen-activated protein kinase activation. AB - Focal adhesion kinase (FAK) is a cytoplasmic tyrosine kinase involved in integrin mediated signal transduction pathway. In this report, we describe that the treatment of hepatocyte growth factor (HGF) stimulates a significant increase in the tyrosine phosphorylation of FAK in human embryonic kidney 293 cells. This stimulation is independent of cell adhesion or the integrity of the actin cytoskeleton, suggesting potentially different mechanisms by which the HGF receptors and integrins regulate the tyrosine phosphorylation of FAK. Our results also suggest that the activation of Src upon HGF stimulation is likely to be one, if not the only, of the mechanisms responsible for the HGF-induced tyrosine phosphorylation of FAK. Furthermore, we showed that a mutation in the Grb2 binding site Tyr-925 of FAK partially abolishes its increase in HGF-induced phosphorylation. Finally, we demonstrated that HGF stimulates the association of FAK with Grb2 in vitro and in intact cells and provided evidence that FAK might contribute to the activation of mitogen-activated protein kinase through Ras in HGF signaling by functioning as an adapter molecule. PMID- 9748250 TI - AKAP15 anchors cAMP-dependent protein kinase to brain sodium channels. AB - The voltage-sensitive sodium channel is regulated by cAMP-dependent protein kinase (PKA) phosphorylation. Using purified preparations of rat brain sodium channels, we have shown that the alpha subunit was phosphorylated by a co purifying protein kinase. The co-purifying kinase was stimulated by cAMP and phosphorylated PKA substrate peptides. Both the regulatory and catalytic subunits of PKA were detected by immunoblotting in purified sodium channel preparations. Bound PKA was immunoprecipitated with anti-SP19 antibodies directed against the sodium channel alpha subunit. PKA bound to sodium channels phosphorylated the sodium channel alpha subunit on the same four serine residues as observed with exogenously added PKA, indicating that association with the sodium channel does not restrict the sites of phosphorylation. Analysis of proteins with high affinity for the type II alpha regulatory subunit of PKA in a gel overlay assay identified a 15-kDa cAMP-dependent protein kinase-anchoring protein (AKAP) in these preparations. Determination of its amino acid sequence by mass spectrometry revealed two peptides identical to AKAP15, a recently described AKAP that targets PKA to skeletal muscle calcium channels. The co-purifying AKAP was also immunoreactive with antibodies generated against AKAP15, and antibodies directed against AKAP15 co-immunoprecipitated the sodium channel. Our results indicate that PKA is bound to brain sodium channels through interaction with AKAP15. Association of AKAP15 with both skeletal muscle calcium channels and brain sodium channels suggests that it may have broad specificity in targeting PKA to ion channels for regulation. PMID- 9748251 TI - Molecular interaction between the Fyn-associated protein SKAP55 and the SLP-76 associated phosphoprotein SLAP-130. AB - It has previously been reported that in resting T-lymphocytes the protein tyrosine kinase p59 constitutively co-precipitates with four phosphoproteins of 43, 55, 85, and 120 kDa, respectively. We have recently cloned the 55-kDa protein that was termed Src kinase-associated phosphoprotein of 55 kDa (SKAP55). Here we demonstrate that the recently characterized SH2-domain-containing leukocyte protein 76-associated phosphoprotein of 130 kDa (SLAP-130) is one of the components of the Fyn complex and that it also co-precipitates with SKAP55 in human T-cells. We establish that SKAP55 and SLAP-130 associate with each other when both molecules are co-expressed in COS cells. By co-transfection of truncated mutants of SKAP55 and SLAP-130 as well as by using the two-hybrid selection system, we further demonstrate that the association between SLAP-130 and SKAP55 is direct and involves the Src homology 3 domain of SKAP55 and the proline-rich sequence of SLAP-130. PMID- 9748252 TI - Cardiac fibroblasts arrest at the G1/S restriction point in response to interleukin (IL)-1beta. Evidence for IL-1beta-induced hypophosphorylation of the retinoblastoma protein. AB - Although responsible for only approximately one-third of the overall myocardial mass, the interstitial fibroblasts of the heart serve a fundamental role in establishing the functional integrity of myocardium and are the major source of myocardial extracellular matrix production. Their importance in clinical medicine is underscored by the observation that fibroblast numbers increase in response to several pathologic circumstances that are associated with an increase in extracellular matrix production, such as long standing hypertension and myocardial injury/infarction. Up to the present time, however, there has been little information available on either the kinetics of the cardiac fibroblast cell cycle, or the fundamental mechanisms that regulate its entry into and exit from the cell cycle. Previous work from our laboratory examining the effects of interleukin (IL)-1beta on myocardial growth and gene expression in culture indicated that cardiac fibroblasts have a diminished capacity to synthesize DNA in response to mitogen in the presence of this cytokine. The mechanism of IL 1beta action was not clear, however, and could have resulted from action at several different points in the cell cycle. The investigations described in this report indicate that IL-1beta exerts its effect on the fibroblast cell cycle at multiple levels through altering the expression of cardiac fibroblast cyclins, cyclin-dependent kinases, and their inhibitors, which ultimately affect the phosphorylation of the retinoblastoma gene product. PMID- 9748253 TI - Superoxide generation from endothelial nitric-oxide synthase. A Ca2+/calmodulin dependent and tetrahydrobiopterin regulatory process. AB - It has been previously shown that besides synthesizing nitric oxide (NO), neuronal and inducible NO synthase (NOS) generates superoxide (O-2) under conditions of L-arginine depletion. However, there is controversy regarding whether endothelial NOS (eNOS) can also produce O-2. Moreover, the mechanism and control of this process are not fully understood. Therefore, we performed electron paramagnetic resonance spin-trapping experiments to directly measure and characterize the O-2 generation from purified eNOS. With the spin trap 5,5 dimethyl-1-pyrroline-N-oxide (DMPO), prominent signals of O-2 adduct, DMPO-OOH, were detected from eNOS in the absence of added tetrahydrobiopterin (BH4), and these were quenched by superoxide dismutase. This O-2 formation required Ca2+/calmodulin and was blocked by the specific NOS inhibitor N-nitro-L-arginine methyl ester (L-NAME) but not its non-inhibitory enantiomer D-NAME. A parallel process of Ca2+/calmodulin-dependent NADPH oxidation was observed which was also inhibited by L-NAME but not D-NAME. Pretreatment of the enzyme with the heme blockers cyanide or imidazole also prevented O-2 generation. BH4 exerted dose dependent inhibition of the O-2 signals generated by eNOS. Conversely, in the absence of BH4 L-arginine did not decrease this O-2 generation. Thus, eNOS can also catalyze O-2 formation, and this appears to occur primarily at the heme center of its oxygenase domain. O-2 synthesis from eNOS requires Ca2+/calmodulin and is primarily regulated by BH4 rather than L-arginine. PMID- 9748254 TI - Random sequence mutagenesis and resistance to 5-fluorouridine in human thymidylate synthases. AB - Thymidylate synthase (TS) catalyzes the methylation of dUMP to dTMP and is the target for the widely used chemotherapeutic agent 5-fluorouracil. We used random sequence mutagenesis to replace 13 codons within the active site of TS and obtain variants that are resistant to 5-fluorodeoxyuridine (5-FdUR). The resulting random library was selected for its ability to complement a TS-deficient Escherichia coli strain, and sequence analysis of survivors found multiple substitutions to be tolerable within the targeted region. An independent selection of the library was carried out in the presence of 5-FdUR, resulting in a more limited spectrum of mutations. One specific mutation, C199L, was observed in more than 46% of 5-FdUR-resistant clones. A 5-FdUR-resistant triple mutant, A197V/L198I/C199F, was purified to apparent homogeneity. Kinetic studies with the substrate dUMP indicate that this mutant is similar to the wild type in regards to kcat and Km values for dUMP and the cosubstrate CH2H4-folate. In contrast, equilibrium binding studies with the inhibitor, FdUMP, demonstrate that the dissociation constant (Kd) for FdUMP binding into the ternary complex was 20-fold higher than values obtained for the wild-type enzyme. This 5-FdUMP-resistant mutant, or others similarly selected, is a candidate for use in gene therapy to render susceptible normal cells resistant to the toxic effects of systemic 5 fluorouracil. PMID- 9748255 TI - Contributions of the domains of the Bacillus subtilis response regulator Spo0A to transcription stimulation of the spoIIG operon. AB - Spo0A is a response regulator that controls entry into sporulation by specifically stimulating or repressing transcription of critical developmental genes. Response regulators have at least two domains: an output transcription regulation domain and a receiver domain that inhibits the output domain. Phosphorylation of the receiver domain relieves the inhibition. We examined the in vitro transcription activation mechanism for Spo0A, phosphorylated Spo0A (Spo0A approximately P), and a deletion mutant that consists solely of the C terminal output domain (Spo0ABD). Both Spo0A approximately P and Spo0ABD stimulated transcription from the spoIIG promoter 10-fold more efficiently than Spo0A. Spo0A approximately P and Spo0ABD induced DNA denaturation by RNA polymerase in the -10 recognition region, whereas Spo0A did not. DNase I footprint assays revealed that phosphorylation enhanced binding of intact Spo0A to the 0A boxes, while the binding of Spo0ABD was similar to that of Spo0A. Thus, activation of Spo0A by phosphorylation is not primarily due to enhanced DNA binding. The presence of a phosphorylated N terminus increased the stability of the ternary complex at the spoIIG promoter. We propose that the primary effect of phosphorylation is to expose an RNA polymerase interaction domain to promote transcription from PspoIIG. PMID- 9748256 TI - The low density lipoprotein receptor active conformation of apolipoprotein E. Helix organization in n-terminal domain-phospholipid disc particles. AB - Lipid association is a prerequisite for receptor interactions of apolipoprotein E (apoE). Disc complexes of the N-terminal 22-kDa apoE3 receptor binding domain and dimyristoylphosphatidylcholine display full receptor binding activity. Studies have been performed to characterize conformational adaptations of the globular, lipid-free four-helix bundle structure that culminate in stable association of its amphipathic alpha-helices with a lipid surface. Helix-lipid interactions in bilayer disc complexes can conceivably adopt two orientations: parallel or perpendicular to the phospholipid acyl chains. Evidence based on infrared dichroism, geometrical arguments, and x-ray crystallography support the view that defined helical segments in the four-helix bundle realign upon lipid association, orienting perpendicular to the phospholipid fatty acyl chains, circumscribing the bilayer disc. Thus, it is likely that paired helical segments align in tandem, presenting a convex receptor-active surface. PMID- 9748257 TI - Identification of a Gialpha binding site on type V adenylyl cyclase. AB - The stimulatory G protein alpha subunit Gsalpha binds within a cleft in adenylyl cyclase formed by the alpha1-alpha2 and alpha3-beta4 loops of the C2 domain. The pseudosymmetry of the C1 and C2 domains of adenylyl cyclase suggests that the homologous inhibitory alpha subunit Gialpha could bind to the analogous cleft within C1. We demonstrate that myristoylated guanosine 5'-3-O-(thio)triphosphate Gialpha1 forms a stable complex with the C1 (but not the C2) domain of type V adenylyl cyclase. Mutagenesis of the membrane-bound enzyme identified residues whose alteration either increased or substantially decreased the IC50 for inhibition by Gialpha1. These mutations suggest binding of Gialpha within the cleft formed by the alpha2 and alpha3 helices of C1, analogous to the Gsalpha binding site in C2. Adenylyl cyclase activity reconstituted by mixture of the C1 and C2 domains of type V adenylyl cyclase was also inhibited by Gialpha. The C1b domain of the type V enzyme contributed to affinity for Gialpha, but the source of C2 had little effect. Mutations in this soluble system faithfully reflected the phenotypes observed with the membrane-bound enzyme. The pseudosymmetrical structure of adenylyl cyclase permits bidirectional regulation of activity by homologous G protein alpha subunits. PMID- 9748258 TI - Characterization of functional domains of an embryonic stem cell coactivator UTF1 which are conserved and essential for potentiation of ATF-2 activity. AB - We have recently cloned a cDNA encoding an embryonic stem cell transcriptional coactivator termed UTF1 from the mouse F9 teratocarcinoma cell line (Okuda, A., Fukushima, A., Nishimoto, M., Orimo, A., Yamagishi, T., Nabeshima, Y., Kuro-o, M., Nabeshima, Y., Boon, K., Keaveney, M., Stunnenberg, H.G., and Muramatsu, M. (1998) EMBO J. 17, 2019-2032). Here we have cloned a cDNA for human UTF1 and identified two highly conserved domains termed conserved domain (CD)1 and CD2. Human UTF1, like that of mouse, binds to ATF-2 and the mutagenesis analyses reveal that the leucine zipper motif within the CD2 of the UTF1 and metal binding motif of ATF-2 are involved in this interaction. The factor also binds to TATA binding protein containing complex. By means of immunoprecipitation analysis, we mapped two domains which are independently able to bind to the complex. Importantly, both domains are located within the conserved domains (one in CD1 and the other in CD2). Furthermore, transient transfection analyses point out the importance of these domains for activating ATF-2. Thus, these results suggest that these two conserved domains identified here play important roles in activating specific transcription at least in part by supporting physical interaction between the upstream factor, ATF-2, and basal transcription machinery. PMID- 9748259 TI - Evidence for a symmetrical requirement for Rab5-GTP in in vitro endosome-endosome fusion. AB - Early endosome fusion, which has been extensively characterized using an in vitro reconstitution assay, is Rab5-dependent. To examine the requirement for Rab5 on both fusion partners, we prepared cytosol and endosomes depleted of Rab5. Unlike control cytosol, Rab5-depleted cytosol was only marginally active in the in vitro endosome fusion. However, fusion could be restored by the addition of wild-type Rab5 or Rab5 D136N, a mutant whose nucleotide specificity favors xanthine over guanine. The addition of Rab5 D136N restored fusion only in the presence of XTP. In the absence of XTP or in the presence of XDP, Rab5 D136N failed to restore fusion. When fusion was carried out with endosomal vesicles depleted of Rab GTPases (by preincubation of vesicles with GDP dissociation inhibitor), together with cytosol immunodepleted of Rab5, fusion was virtually absent. We then used immunodepleted cytosol and GDP dissociation inhibitor-treated vesicles to determine whether Rab5 is required by both fusion partners. Using separate sets of endosomal vesicles, we found that priming both sets of Rab5-depleted vesicles with Rab5 Q79L, a GTPase-defective mutant, substantially stimulated endosome fusion. Priming one set of vesicles with Rab5 Q79L and a second set of vesicles with Rab5 S34N failed to activate fusion. When both sets of Rab5-depleted vesicles were primed with Rab5 D136N supplemented with XTP, endosome fusion was stimulated, similar to that observed with Rab5 Q79L. However, when one set of vesicles was preincubated with Rab5 D136N plus XTP and the second set with Rab5 D136N and XDP, no stimulation of fusion was observed. We conclude that Rab5-GTP is required on both fusion partners for docking and fusion of early endosomes. To confirm the fusion of Rab5-GTP-positive vesicles in vivo, we expressed GFP-Rab5 Q79L in fibroblasts and observed fusion of Rab5-positive vesicles. We failed to record fusion of Rab5-positive vesicles with Rab5-negative vesicles. We conclude that Rab5-GTP is required on both sets of endosomes for fusion in vitro and in living cells. PMID- 9748260 TI - NHE3 kinase A regulatory protein E3KARP binds the epithelial brush border Na+/H+ exchanger NHE3 and the cytoskeletal protein ezrin. AB - Cyclic AMP is a major second messenger that inhibits the brush border Na+/H+ exchanger NHE3. We have previously shown that either of two related regulatory proteins, E3KARP or NHERF, is necessary for the cAMP-dependent inhibition of NHE3. In the present study, we characterized the interaction between NHE3 and E3KARP using in vitro binding assays. We found that NHE3 directly binds to E3KARP and that the entirety of the second PSD-95/Dlg/ZO-1 (PDZ) domain plus the carboxyl-terminal domain of E3KARP are required to bind NHE3. E3KARP binds an internal region within the NHE3 C-terminal cytoplasmic tail, defining a new mode of PDZ domain interaction. Analyses of cellular distribution of NHE3 and E3KARP expressed in PS120 fibroblasts show that NHE3 and E3KARP are co-localized on the plasma membrane, but not in a distinct juxtanuclear compartment in which NHE3 is predominantly expressed. The distributions of NHE3 and E3KARP were not affected by treatment with 8-bromo-cAMP. As shown earlier for the human homolog of NHERF, we also found that the cytoskeletal protein ezrin binds to the carboxyl-terminal domain of E3KARP. These results are consistent with the possibility that E3KARP and NHERF may function as scaffold proteins that bind to both NHE3 and ezrin. Since ezrin is a protein kinase A anchoring protein, we suggest that the scaffolding function of E3KARP binding to both ezrin and NHE3 localizes cAMP dependent protein kinase in the vicinity of the cytoplasmic domain of NHE3, which is phosphorylated by elevated cAMP. PMID- 9748261 TI - A role for Saccharomyces cerevisiae fatty acid activation protein 4 in regulating protein N-myristoylation during entry into stationary phase. AB - Saccharomyces cerevisiae contains four known acyl-CoA synthetases (fatty acid activation proteins, Faaps). Faa1p and Faa4p activate exogenously derived fatty acids. Acyl-CoA metabolism plays a critical role in regulating protein N myristoylation by the essential enzyme, myristoyl-CoA:protein N myristoyltransferase (Nmt1p). In this report, we have examined whether Faa1p and Faa4p have distinct roles in affecting protein N-myristoylation as cells transition from growth in rich media to a growth-arrested state during nutrient deprivation (stationary phase). The colony-forming potential of 10 isogenic strains was defined as a function of time spent in stationary phase. These strains contained either a wild type or mutant NMT1 allele, and wild type or null alleles of each FAA. Only the combination of the Nmt mutant (nmt451Dp; reduced affinity for myristoyl-CoA) and loss of Faa4p produced a dramatic loss of colony forming units (CFU). The progressive millionfold reduction in CFU was associated with a deficiency in protein N-myristoylation that first appeared during logarithmic growth, worsened through the post-diauxic phase, and became extreme in stationary phase. Northern and Western blot analyses plus N myristoyltransferase assays showed that Nmt is normally present only during the log and diauxic/post-diauxic periods, indicating that N-myristoylproteins present in stationary phase are "inherited" from these earlier phases. Moreover, FAA4 is the only FAA induced during the critical diauxic/early post-diauxic transition. Although substitution of nmt1-451D for NMT1 results in deficiencies in protein N myristoylation, these deficiencies are modest and limited by compensatory responses that include augmented expression of nmt1-451D and precocious induction of FAA4 in log phase. Loss of Faa4p from nmt1-451D cells severely compromises their capacity to adequately myristoylate Nmt substrates prior to entry into stationary phase since none of the other Faaps are able to functionally compensate for its absence. To identify Nmt1p substrates that may affect maintenance of proliferative potential during stationary phase, we searched the yeast genome for known and putative N-myristoylproteins. Of the 64 genes found, 48 were successfully deleted in NMT1 cells. Removal of any one of the following nine substrates produced a loss of CFU similar to that observed in nmt1 451Dfaa4Delta cells: Arf1p, Arf2p, Sip2p, Van1p, Ptc2p, YBL049W (homology to Snf7p), YJR114W, YKR007W, and YMR077C. These proteins provide opportunities to further define the molecular mechanisms that regulate survival during stationary phase. PMID- 9748262 TI - Homeodomain-interacting protein kinases, a novel family of co-repressors for homeodomain transcription factors. AB - A novel family of cofactors that differentially interact with homeoproteins have been identified via a yeast two-hybrid screen. The proteins contain a conserved protein kinase domain that is separated from a domain that interacts with homeoproteins and hence are termed homeodomain-interacting protein kinases (HIPKs): HIPK1, HIPK2, and HIPK3. We show that HIPKs are nuclear kinases using GFP-HIPK fusion constructs. The DNA binding activity of the NK-3 homeoprotein is greatly enhanced by HIPK2, but this effect is independent of its phosphorylation by HIPK2. In cultured cells, HIPKs localize to nuclear speckles and potentiate the repressor activities of NK homeoproteins. The co-repressor activity of HIPKs depends on both its homeodomain interaction domain and a co-repressor domain that maps to the N terminus. Thus, HIPKs represent a heretofore undescribed family of co-repressors for homeodomain transcription factors. PMID- 9748263 TI - UDP-galactose:ceramide galactosyltransferase is a class I integral membrane protein of the endoplasmic reticulum. AB - UDP-galactose:ceramide galactosyltransferase (CGalT) transfers UDP-galactose to ceramide to form the glycosphingolipid galactosylceramide. Galactosylceramide is the major constituent of myelin and is also highly enriched in many epithelial cells, where it is thought to play an important role in lipid and protein sorting. Although the biochemical pathways of glycosphingolipid biosynthesis are relatively well understood, the localization of the enzymes involved in these processes has remained controversial. We here have raised antibodies against CGalT and shown by immunocytochemistry on ultrathin cryosections that the enzyme is localized to the endoplasmic reticulum and nuclear envelope but not to the Golgi apparatus or the plasma membrane. In pulse-chase experiments, we have observed that newly synthesized CGalT remains sensitive to endoglycosidase H, confirming the results of the morphological localization experiments. In protease protection assays, we show that the largest part of the protein, including the amino terminus, is oriented toward the lumen of the endoplasmic reticulum. CGalT enzyme activity required import of UDP-galactose into the lumen of the endoplasmic reticulum by a UDP-galactose translocator that is present in the Golgi apparatus of CHO cells but absent in CHOlec8 cells. Finally, we show that CGalT activity previously observed in Golgi membrane fractions in vitro, in the absence of UDP-glucose, is caused by UDP-glucose:ceramide glucosyltransferase. Therefore all galactosylceramide synthesis occurs by CGalT in vivo in the lumen of the endoplasmic reticulum. PMID- 9748264 TI - Tarantula hemocyanin shows phenoloxidase activity. AB - An enzyme generally catalyzes one well defined reaction with high specificity and efficiency. We report here in contrast that the copper protein hemocyanin of the tarantula Eurypelma californicum exhibits two different functions. These occur at the same active site. While hemocyanin usually is an oxygen carrier, its function can be transformed totally to monophenoloxidase and o-diphenoloxidase activity after limited proteolysis with trypsin or chymotrypsin. N-acetyldopamine (NADA) is more effectively oxidized than L-dopa or dopamine. This irreversible functional switch of tarantula hemocyanin function is limited to the two subunits b and c of its seven subunit types. A conserved phenylalanine in the hemocyanin molecule acts as a placeholder for other substrates that are phenylalanine derivatives. The proteolytic cleavage removes an N-terminal fragment, including the critical phenylalanine residue, which opens an entrance for substrates. Therefore no new arrangement of the active site, with its two copper atoms and the mu - eta2:eta2 bound O2 molecule, is necessary to develop the catalytic function. PMID- 9748265 TI - Sequence characteristics, subcellular localization, and substrate specificity of DYRK-related kinases, a novel family of dual specificity protein kinases. AB - DYRK1 is a dual specificity protein kinase presumably involved in brain development. Here we show that the kinase belongs to a new family of protein kinases comprising at least seven mammalian isoforms (DYRK1A, DYRK1B, DYRK1C, DYRK2, DYRK3, DYRK4A, and DYRK4B), the yeast homolog Yak1p, and the Drosophila kinase minibrain (MNB). In rat tissues, DYRK1A is expressed ubiquitously, whereas transcripts for DYRK1B, DYRK2, DYRK3, and DYRK4 were detected predominantly in testes of adult but not prepuberal rats. By fluorescence microscopy and subcellular fractionation, a green fluorescent protein (GFP) fusion protein of DYRK1A was found to accumulate in the nucleus of transfected COS-7 and HEK293 cells, whereas GFP-DYRK2 was predominantly detected in the cytoplasm. DYRK1A exhibited a punctate pattern of GFP fluorescence inside the nucleus and was co purified with the nuclear matrix. Analysis of GFP-DYRK1A deletion constructs showed that the nuclear localization of DYRK1A was mediated by its nuclear targeting signal (amino acids 105-139) but that its characteristic subnuclear distribution depended on additional N-terminal elements (amino acids 1-104). When expressed in Escherichia coli, DYRK1A, DYRK2, DYRK3, MNB, and Yak1p catalyzed their autophosphorylation on tyrosine residues. The kinases differed in their substrate specificity in that DYRK2 and DYRK3, but not DYRK1A and MNB, catalyzed phosphorylation of histone H2B. The heterogeneity of their subcellular localization and substrate specificity suggests that the kinases are involved in different cellular functions. PMID- 9748266 TI - Gelatinase B/lacZ transgenic mice, a model for mapping gelatinase B expression during developmental and injury-related tissue remodeling. AB - Matrix metalloproteinases (MMPs) drive normal tissue remodeling and are implicated in a wide range of pathologies. Although MMP activity is controlled at multiple levels, the primary regulation of MMP activity is transcriptional. The transcriptional promoter elements required for MMP gene expression in cultured cells have been defined, but this has not been extended to the in vivo situation. In this paper, we show that the DNA sequences between -522 and +19 of the rabbit gelatinase B gene (MMP-9) (as characterized in the transgenic mouse line 3445) constitute a minimal promoter that drives appropriate developmental and injury induced reporter gene expression in transgenic mice. We further show that the expression and activity of three transcription factors (NF-kappaB, AP-2, and Sp1) that control the activity of the gelatinase B promoter are selectively induced in the epithelium migrating to heal a wound. Although promoter activity parallels expression of the endogenous gene in cell cultures, we show by several criteria that cell cultures cannot model many aspects of promoter regulation in vivo. This study reveals that the transgenic mouse line 3445 might be a useful model for investigating the regulation of gelatinase B expression in vivo and for identifying and characterizing new drugs that can control gelatinase B gene transcription. PMID- 9748267 TI - The medium subunits of adaptor complexes recognize distinct but overlapping sets of tyrosine-based sorting signals. AB - Tyrosine-based sorting signals conforming to the motif YXXO (Y is tyrosine, X is any amino acid, and O is an amino acid with a bulky hydrophobic side chain (leucine, isoleucine, phenylalanine, methionine, valine)) interact with the medium (mu) subunits of clathrin adaptor (AP) complexes. We have analyzed the selectivity of interaction between YXXO signals and the mu1, mu2, and mu3 (A or B) subunits of the AP-1, AP-2, and AP-3 complexes, respectively, by screening a combinatorial XXXYXXO library using the yeast two-hybrid system. All the medium subunits were found to prefer proline at position Y+2, suggesting that YXXO signals are stabilized by a bend in the polypeptide backbone. Other than for this common preference, each medium subunit favored specific sets of residues at the X and O positions; these preferences were consistent with the proposed roles of the different adaptor complexes in rapid endocytosis and lysosomal targeting. A considerable specificity overlap was also revealed by these analyses, suggesting that additional factors, such as the context of the signals, must be important determinants of recognition. PMID- 9748268 TI - Expression of Id1 results in apoptosis of cardiac myocytes through a redox dependent mechanism. AB - We have constructed a recombinant adenovirus (Ad.Id1) that allows for efficient expression of the helix-loop-helix protein Id1. After infection with Ad.Id1, neonatal cardiac myocytes display a significant reduction in viability, which was proportional to the level of Id1 expression. A similar effect was observed in adult myocytes. Morphological and biochemical assays demonstrated that Id1 expression resulted in myocyte apoptosis. In contrast, expression of Id1 in endothelial cells, vascular smooth muscle cells, or fibroblasts did not affect the viability of these cells. Along with the induction of apoptosis, the expression of Id1 in neonatal cardiac myocytes resulted in an increase in the level of intracellular reactive oxygen species. The source of these reactive oxygen species appears to be the mitochondria. Reducing the ambient oxygen concentration or treatment with a cell-permeant H2O2 scavenger prevented Id1 stimulated apoptosis in cardiac myocytes. These results suggest that the expression of Id1 leads to the induction of apoptosis in cardiac myocytes through a redox-dependent mechanism. PMID- 9748269 TI - Molecular cloning and characterization of TIEG2 reveals a new subfamily of transforming growth factor-beta-inducible Sp1-like zinc finger-encoding genes involved in the regulation of cell growth. AB - Sp1-like zinc finger transcription factors are involved in the regulation of cell growth and differentiation. Recent evidence demonstrating that mammalian cells express novel, yet uncharacterized, Sp1-like proteins has stimulated a search for new members of this family. We and others have recently reported that the transforming growth factor (TGF)-beta-regulated gene TIEG encodes a new Sp1-like protein that inhibits cell growth in cultured cells. Here we report the identification, nuclear localization, DNA binding activity, transcriptional repression activity, and growth inhibitory effects of TIEG2, a novel TGF-beta inducible gene related to TIEG. TIEG2 is ubiquitously expressed in human tissues, with an enrichment in pancreas and muscle. TIEG2 shares 91% homology with TIEG1 within the zinc finger region and 44% homology within the N terminus. Biochemical characterization reveals that TIEG2 is a nuclear protein, which, as predicted from the primary structure, specifically binds to an Sp1-like DNA sequence in vitro and can repress a promoter containing Sp1-like binding sites in transfected Chinese hamster ovary epithelial cells. Furthermore, functional studies using [3H]thymidine uptake and MTS (3-(4, 3-dimethyltiazol-2-yl)-5-(3 carboxymethoxyphenyl)-2-(4-su lfophenyl)-2 H-tetrazolium) assays demonstrate that the overexpression of TIEG2 in Chinese hamster ovary cells inhibits cell proliferation. Thus, TIEG2, together with TIEG1, defines a new subfamily of TGF beta-inducible Sp1-like proteins involved in the regulation of cell growth. PMID- 9748271 TI - Cellular folates prevent polyglutamation of 5, 10-dideazatetrahydrofolate. A novel mechanism of resistance to folate antimetabolites. AB - Mouse L1210 cell variants were selected for resistance to 5, 10 dideazatetrahydrofolate, a potent inhibitor of the first folate-dependent enzyme in de novo purine synthesis, glycinamide ribonucleotide formyltransferase. The drug-resistant phenotype selected was conditional to the folate compound used to support growth: grown on folic acid cells were 400-fold resistant, whereas they were 2.5-fold more sensitive to 5,10-dideazatetrahydrofolate than wild-type L1210 cells when grown on folinic acid. In folic acid-containing media, polyglutamation of 5, 10-dideazatetrahydrofolate was markedly reduced, yet folylpolyglutamate synthetase activity was not different from that in parental L1210 cells. Resistance was due to two changes in membrane transport: a minor increase in the Km for 5, 10-dideazatetrahydrofolate influx, and a major increase in folic acid transport. Enhanced folic acid transport resulted in an expanded cellular content of folates which blocked polyglutamation of 5,10-dideazatetrahydrofolate. We propose that polyglutamation of 5,10-dideazatetrahydrofolate is limited by feedback inhibition by cellular folates on folylpolyglutamate synthetase, an effect which reflects a mechanism in place to control the level of cellular folates. Although the primary alteration causative of resistance is different from those reported previously, all 5, 10-dideazatetrahydrofolate resistance phenotypes result in decreased drug polyglutamation, reflecting the centrality of this reaction to the action of 5,10-dideazatetrahydrofolate. PMID- 9748270 TI - Two forms of collagen XVII in keratinocytes. A full-length transmembrane protein and a soluble ectodomain. AB - The cDNA sequence of human collagen XVII predicts an unusual type II transmembrane protein, but a biochemical characterization of this structure has not been accomplished yet. Using domain-specific antibodies against recombinant collagen XVII fragments, we identified two molecular forms of the collagen in human skin and epithelial cells. Full-length collagen XVII appeared as a homotrimeric transmembrane molecule of three 180-kDa alpha1(XVII) chains. The globular intracellular domain was disulfide-linked, and the N-glycosylated extracellular domain of three 120-kDa polypeptides was triple-helical at physiological temperatures. A second, soluble form of collagen XVII in keratinocyte culture media was recognized with antibodies to the ectodomain, but not the endodomain. The soluble form exhibited molecular properties of the collagen XVII ectodomain: a triple-helical, N-glycosylated molecule of three 120 kDa polypeptides. Northern blot analysis with probes spanning either the distal 5'or the distal 3' end of the collagen XVII cDNA revealed an identical 6-kb mRNA, suggesting that both the 180- and 120-kDa polypeptides were translated from the same mRNA, and that the 120-kDa polypeptide was generated post-translationally. In concert, keratinocytes harboring a homozygous nonsense mutation in the COL17A1 gene synthesized neither the 180-kDa alpha1(XVII) chain nor the 120-kDa polypeptide. Finally, treatment of normal keratinocytes with a synthetic inhibitor of furin proprotein convertases, decanoyl-RVKR-chloromethyl ketone, prevented the generation of the 120-kDa polypeptide. These data strongly suggest that the soluble 120-kDa polypeptide represents a specifically cleaved ectodomain of collagen XVII, generated through furin-mediated proteolytic processing. Thus, collagen XVII is not only an unusual type II transmembrane collagen, but the first collagen with a specifically processed, soluble triple-helical ectodomain. PMID- 9748272 TI - Mutations in the reduced folate carrier gene which confer dominant resistance to 5,10-dideazatetrahydrofolate. AB - L1210/D3 mouse leukemia cells are resistant to 5, 10-dideazatetrahydrofolate due to expansion of cellular folate pools which block polyglutamation of the drug (Tse, A., and Moran, R. G. (1998) J. Biol. Chem. 273, 25944-25952). These cells were found to have two point mutations in the reduced folate carrier (RFC), resulting in a replacement of isoleucine 48 by phenylalanine and of tryptophan 105 by glycine. Each mutation contributes to the resistance phenotype. Genomic DNA from resistant cells contained both the wild-type and mutant alleles, but wild-type message was not detected. Folic acid was a much better substrate, and 5 formyltetrahydrofolate was a poorer substrate for transport in L1210/D3 cells relative to L1210 cells. Enhanced transport of folic acid was due to a marked, approximately 20-fold, decrease in the influx Km. Influx of methotrexate and 5,10 dideazatetrahydrofolate were minimally altered. Transfection of mutated rfc cDNA into RFC-null L1210/A cells produced the substrate specificity and 5, 10 dideazatetrahydrofolate resistance observed in the L1210/D3 line. Transfection of the mutant cDNA into wild-type cells also conferred resistance to 5,10 dideazatetrahydrofolate. We conclude that the I48F and W105G mutations in RFC caused resistance to 5, 10-dideazatetrahydrofolate, that the region of the RFC protein near these two positions defines the substrate-binding site, that the wild-type allele was silenced during the multistep development of resistance, and that this mutant phenotype represents a genetically dominant trait. PMID- 9748273 TI - Insulin-like growth factor-I inhibits the stress-activated protein kinase/c-Jun N terminal kinase. AB - The pathways involved in the cellular responses to the insulin-like growth factors (IGFs) are numerous and vary according to cell type. Following activation of the IGF-I receptor, the mitogen-activated protein kinase and phosphatidylinositide 3'-kinase (PI3'K) pathways are activated and result in cellular proliferation and inhibition of apoptosis. In this study, we analyzed the IGF-I effect on the stress-activated protein kinase/c-Jun N-terminal kinase (JNK) activity using human embryonic kidney 293 cells, 293 cells transiently expressing hemagglutinin-JNK, and 293 cells stably expressing a hemagglutinin-JNK transgene. In all cell types, endogenous or transfected JNK activity was strongly stimulated by anisomycin or tumor necrosis factor-alpha, and 10 nM IGF-I pretreatment suppressed the induced JNK activity. To determine whether the effect of IGF-I on JNK activity involves the mitogen-activated protein kinase or PI3'K pathway, we used the specific MEK1 inhibitor PD098059 and the PI3'K inhibitor LY 294002. PD098059 did not alter the IGF-I suppressive effect on stressor-induced JNK activity, but LY 294002 suppressed the IGF-I effect. Moreover, in transiently transfected parental 293 cells expressing dominant-negative Akt, anisomycin increased JNK activity was not suppressed by pretreatment with IGF-I. Our results demonstrate that the action of IGF-I on JNK in these cells is via PI3'K and Akt. PMID- 9748274 TI - Identification of PKDL, a novel polycystic kidney disease 2-like gene whose murine homologue is deleted in mice with kidney and retinal defects. AB - Polycystin-1 and polycystin-2 are the products of PKD1 and PKD2, genes that are mutated in most cases of autosomal dominant polycystic kidney disease. Polycystin 2 shares approximately 46% homology with pore-forming domains of a number of cation channels. It has been suggested that polycystin-2 may function as a subunit of an ion channel whose activity is regulated by polycystin-1. Here we report the identification of a human gene, PKDL, which encodes a new member of the polycystin protein family designated polycystin-L. Polycystin-L has 50% amino acid sequence identity and 71% homology to polycystin-2 and has striking sequence and structural resemblance to the pore-forming alpha1 subunits of Ca2+ channels, suggesting that polycystin-L may function as a subunit of an ion channel. The full-length transcript of PKDL is expressed at high levels in fetal tissues, including kidney and liver, and down-regulated in adult tissues. PKDL was assigned to 10q24 by fluorescence in situ hybridization and is linked to D10S603 by radiation hybrid mapping. There is no evidence of linkage to PKDL in six ADPKD families that are unlinked to PKD1 or PKD2. The mouse homologue of PKDL is deleted in Krd mice, a deletion mutant with defects in the kidney and eye. We propose that PKDL is an excellent candidate for as yet unmapped cystic diseases in man and animals. PMID- 9748275 TI - Catabolism of phenylacetic acid in Escherichia coli. Characterization of a new aerobic hybrid pathway. AB - The paa cluster of Escherichia coli W involved in the aerobic catabolism of phenylacetic acid (PA) has been cloned and sequenced. It was shown to map at min 31.0 of the chromosome at the right end of the mao region responsible for the transformation of 2-phenylethylamine into PA. The 14 paa genes are organized in three transcription units: paaZ and paaABCDEFGHIJK, encoding catabolic genes; and paaXY, containing the paaX regulatory gene. The paaK gene codes for a phenylacetyl-CoA ligase that catalyzes the activation of PA to phenylacetyl-CoA (PA-CoA). The paaABCDE gene products, which may constitute a multicomponent oxygenase, are involved in PA-CoA hydroxylation. The PaaZ protein appears to catalyze the third enzymatic step, with the paaFGHIJ gene products, which show significant similarity to fatty acid beta-oxidation enzymes, likely involved in further mineralization to Krebs cycle intermediates. Three promoters, Pz, Pa, and Px, driven the expression of genes paaZ, paaABCDEFGHIJK, and paaX, respectively, have been identified. The Pa promoter is negatively controlled by the paaX gene product. As PA-CoA is the true inducer, PaaX becomes the first regulator of an aromatic catabolic pathway that responds to a CoA derivative. The aerobic catabolism of PA in E. coli represents a novel hybrid pathway that could be a widespread way of PA catabolism in bacteria. PMID- 9748276 TI - The CC chemokine monocyte chemotactic peptide-1 activates both the class I p85/p110 phosphatidylinositol 3-kinase and the class II PI3K-C2alpha. AB - The cellular effects of MCP-1 are mediated primarily by binding to CC chemokine receptor-2. We report here that MCP-1 stimulates the formation of the lipid products of phosphatidylinositol (PI) 3-kinase, namely phosphatidylinositol 3,4 bisphosphate and phosphatidylinositol 3,4,5-trisphosphate (PI 3,4,5-P3) in THP-1 cells that can be inhibited by pertussis toxin but not wortmannin. MCP-1 also stimulates an increase in the in vitro lipid kinase activity present in immunoprecipitates of the class 1A p85/p110 heterodimeric PI 3-kinase, although the kinetics of activation were much slower than observed for the accumulation of PI 3,4,5-P3. In addition, this in vitro lipid kinase activity was inhibited by wortmannin (IC50 = 4.47 +/- 1.88 nM, n = 4), and comparable concentrations of wortmannin also inhibited MCP-stimulated chemotaxis of THP-1 cells (IC50 = 11.8 +/- 4.2 nM, n = 4), indicating that p85/p110 PI 3-kinase activity is functionally relevant. MCP-1 also induced tyrosine phosphorylation of three proteins in these cells, and a fourth tyrosine-phosphorylated protein co-precipitates with the p85 subunit upon MCP-1 stimulation. In addition, MCP-1 stimulated lipid kinase activity present in immunoprecipitates of a class II PI 3-kinase (PI3K-C2alpha) with kinetics that closely resembled the accumulation of PI 3,4,5-P3. Moreover, this MCP-1-induced increase in PI3K-C2alpha activity was insensitive to wortmannin but was inhibited by pertussis toxin pretreatment. Since this mirrored the effects of these inhibitors on MCP-1-stimulated increases in D-3 phosphatidylinositol lipid accumulation in vivo, these results suggest that activation of PI3K-C2alpha rather than the p85/p110 heterodimer is responsible for mediating the in vivo formation of D-3 phosphatidylinositol lipids. These data demonstrate that MCP-1 stimulates protein tyrosine kinases as well as at least two separate PI 3-kinase isoforms, namely the p85/p110 PI 3-kinase and PI3K C2alpha. This is the first demonstration that MCP-1 can stimulate PI 3-kinase activation and is also the first indication of an agonist-induced activation of the PI3K-C2alpha enzyme. These two events may play important roles in MCP-1 stimulated signal transduction and biological consequences. PMID- 9748277 TI - A conserved element in the serine protease domain of complement factor B. AB - Factor B and C2 are serine proteases that carry the catalytic sites of the complement C3 and C5 convertases. Their protease domains are activated by conformational changes that occur during convertase assembly and are deactivated upon convertase dissociation. Factor B and C2 share an 8-amino acid conserved sequence near their serine protease termini that is not seen in other serine proteases. To determine its importance, 24 factor B mutants were generated, each with a single amino acid substitution in this region. Whereas most mutants were functionally neutral, all five different substitutions of aspartic acid 715 and one phenylalanine 716 substitution severely reduced hemolytic activity. Several aspartic acid 715 mutants permitted the steps of convertase assembly including C3b-dependent factor D-mediated cleavage and activation of the high affinity C3b binding site, but the resulting complexes did not cleave C3. Given that factor B and C2 share the same biological substrates and that part of the trypsin-like substrate specificity region is not apparent in either protein, we propose that the conserved region plays a critical role in the conformational regulation of the catalytic site and could offer a highly specific target for the therapeutic inhibition of complement. PMID- 9748278 TI - Glucosylceramide synthase inhibitor inhibits the action of nerve growth factor in PC12 cells. AB - Previous studies have shown that the ceramide analogue, D-threo-1-phenyl-2 decanoylamin-3-morpholino-propanol (D-PDMP), inhibits glucosylceramide synthase and thus leads to extensive depletion of glycosphingolipids derived from glucosyl ceramide. Our previous studies have shown that cholera toxin B subunit, which specifically binds to the cell surface ganglioside GM1, and GM1 itself can enhance the action of nerve growth factor (NGF) in responsive cells by enhancing the NGF-induced autophosphorylation of the high affinity NGF receptor, Trk. Using D-PDMP, we examined the effects of the inhibition of the biosynthesis of glycosphingolipids on intracellular NGF signaling pathway. D-PDMP was found to inhibit NGF-induced neurite outgrowth of PC12 cells. Moreover, D-PDMP clearly inhibited NGF-induced autophosphorylation of Trk and prevented the activation of phosphatidylinositol 3-kinase and mitogen-activated protein kinase, downstream targets of Trk-initiated intracellular protein kinase cascades. These effects of D-PDMP were abolished by the addition of GM1 but not by the addition of other ganglioside subspecies to the culture medium. Furthermore, the effect of D-PDMP seemed to be specific for the Trk receptor, because intracellular signaling pathway of epidermal growth factor was not affected by D-PDMP. Dimethylsphingosine and the cell-permeable analogue, C2-ceramide, did not show such a strong inhibitory effect on neurite outgrowth or on the autophosphorylation of Trk. The present results and our previous observations clearly demonstrate that Trk requires endogenous gangliosides, especially GM1, for its normal function in mediating the neurotrophic activity of NGF at least in PC12 cells. PMID- 9748279 TI - RGSZ1, a Gz-selective regulator of G protein signaling whose action is sensitive to the phosphorylation state of Gzalpha. AB - Regulators of G protein signaling (RGS) are a family of proteins that attenuate the activity of the trimeric G proteins. RGS proteins act as GTPase-activating proteins (GAPs) for the alpha subunits of several trimeric G proteins, much like the GAPs that regulate the activity of monomeric G proteins such as Ras. RGS proteins have been cloned from many eukaryotes, and those whose biochemical activity has been characterized regulate the members of the Gi family of G proteins; some forms can also act on Gq proteins. In an ongoing effort to elucidate the role of Gzalpha in cell signaling, the yeast two-hybrid system was employed to identify proteins that could interact with a mutationally activated form of Gzalpha. A novel RGS, termed RGSZ1, was identified that is most closely related to two existing RGS proteins termed RetRGS1 and GAIP. Northern blot analysis revealed that expression of RGSZ1 was limited to brain, and expression was particularly high in the caudate nucleus. Biochemical characterization of recombinant RGSZ1 protein revealed that RGSZ1 was indeed a GAP and, most significantly, showed a marked preference for Gzalpha over other members of the Gialpha family. Phosphorylation of Gzalpha by protein kinase C, an event known to occur in cells and that was previously shown to influence alpha-betagamma interactions of Gz, rendered the G protein much less susceptible to RGSZ1 action. PMID- 9748281 TI - Identification of the rat adapter Grb14 as an inhibitor of insulin actions. AB - We cloned by interaction with the beta-subunit of the insulin receptor the rat variant of the human adapter Grb14 (rGrb14). rGrb14 is specifically expressed in rat insulin-sensitive tissues and in the brain. The binding of rGrb14 to insulin receptors is insulin-dependent in vivo in Chinese hamster ovary (CHO) cells overexpressing both proteins and importantly, in rat liver expressing physiological levels of proteins. However, rGrb14 is not a substrate of the tyrosine kinase of the receptor. In the two-hybrid system, two domains of rGrb14 can mediate the interaction with insulin receptors: the Src homology 2 (SH2) domain and a region between the PH and SH2 domains that we named PIR (for phosphorylated insulin receptor-interacting region). In vitro interaction assays using deletion mutants of rGrb14 show that the PIR, but not the SH2 domain, is able to coprecipitate insulin receptors, suggesting that the PIR is the major binding domain of rGrb14. The interaction between rGrb14 and the insulin receptors is almost abolished by mutating tyrosine residue Tyr1150 or Tyr1151 of the receptor. The overexpression of rGrb14 in CHO-IR cells decreases insulin stimulation of both DNA and glycogen synthesis. These effects are accompanied by a decrease in insulin-stimulated tyrosine phosphorylation of IRS-1, but insulin receptor autophosphorylation is unaltered. These findings suggest that rGrb14 could be a new downstream signaling component of the insulin-mediated pathways. PMID- 9748280 TI - RGSZ1, a Gz-selective RGS protein in brain. Structure, membrane association, regulation by Galphaz phosphorylation, and relationship to a Gz gtpase-activating protein subfamily. AB - We cloned the cDNA for human RGSZ1, the major Gz-selective GTPase-activating protein (GAP) in brain (Wang, J., Tu, Y., Woodson, J., Song, X., and Ross, E. M. (1997) J. Biol. Chem. 272, 5732-5740) and a member of the RGS family of G protein GAPs. Its sequence is 83% identical to RET-RGS1 (except its N-terminal extension) and 56% identical to GAIP. Purified, recombinant RGSZ1, RET-RGS1, and GAIP each accelerated the hydrolysis of Galphaz-GTP over 400-fold with Km values of approximately 2 nM. RGSZ1 was 100-fold selective for Galphaz over Galphai, unusually specific among RGS proteins. Other enzymological properties of RGSZ1, brain Gz GAP, and RET-RGS1 were identical; GAIP differed only in Mg2+ dependence and in its slightly lower selectivity for Galphaz. RGSZ1, RET-RGS1, and GAIP thus define a subfamily of Gz GAPs within the RGS proteins. RGSZ1 has no obvious membrane-spanning region but is tightly membrane-bound in brain. Its regulatory activity in membranes depends on stable bilayer association. When co reconstituted into phospholipid vesicles with Gz and m2 muscarinic receptors, RGSZ1 increased agonist-stimulated GTPase >15-fold with EC50 <12 nM, but RGSZ1 added to the vesicle suspension was <0.1% as active. RGSZ1, RET-RGS1, and GAIP share a cysteine string sequence, perhaps targeting them to secretory vesicles and allowing them to participate in the proposed control of secretion by Gz. Phosphorylation of Galphaz by protein kinase C inhibited the GAP activity of RGSZ1 and other RGS proteins, providing a mechanism for potentiation of Gz signaling by protein kinase C. PMID- 9748282 TI - Activated thyroglobulin possesses a transforming growth factor-beta activity. AB - Thyroglobulin (Tg), the thyroid hormone precursor, is a major protein component in the thyroid gland and may have other important functions. Here, we show that bovine Tg inhibited 125I-labeled transforming growth factor-beta1 (125I-TGF beta1) binding to cell-surface TGF-beta receptors in mink lung epithelial cells with an IC50 of approximately 300 nM. After disuccinimidyl suberate (DSS) modification, reduction/alkylation, treatment with 8 M urea, 0. 1% SDS, or acidic pH (pH 4-5), Tg exhibited a approximately 5-10-fold increase of 125I-TGF-beta1 binding inhibitory activity with IC50 of approximately 30-60 nM. This inhibitory activity was an intrinsic property of the Tg and could not be segregated from Tg protein by 5% SDS-polyacrylamide gel electrophoresis or by immunoprecipitation using antiserum to Tg. Untreated Tg did not affect DNA synthesis but blocked the TGF-beta-induced inhibition of DNA synthesis in mink lung epithelial cells. After DSS activation, Tg possessed TGF-beta agonist activity and inhibited DNA synthesis of mink lung epithelial cells and rat thyroid cells. The activated Tg also exerted a small but significant TGF-beta agonist activity in transcriptional activation of plasminogen activator inhibitor-1. These results suggest that Tg possesses an authentic TGF-beta activity which can be induced by chemical modifications and treatments with denaturing agents and acidic pH. PMID- 9748283 TI - The DNA replication-related element (DRE)/DRE-binding factor system is a transcriptional regulator of the Drosophila E2F gene. AB - Two mRNA species were observed for the Drosophila E2F (dE2F) gene, differing with regard to the first exons (exon 1-a and exon 1-b), which were expressed differently during development. A single transcription initiation site for mRNA containing exon 1-b was mapped by primer extension analysis and numbered +1. We found three tandemly aligned sequences, similar to the DNA replication-related element (DRE; 5'-TATCGATA), which is commonly required for transcription of genes related to DNA replication and cell proliferation, in the region upstream of this site. Band mobility shift analyses using oligonucleotides containing the DRE related sequences with or without various base substitutions revealed that two out of three DRE-related sequences are especially important for binding to the DRE-binding factor (DREF). On footprinting analysis with Kc cell nuclear extracts and a glutathione S-transferase fusion protein with the N-terminal fragment (1 125 amino acid residues) of DREF, all three DRE-related sequences were found to be protected. Transient luciferase expression assays in Kc cells demonstrated that the region containing the three DRE-related sequences is required for high promoter activity. We have established transgenic lines of Drosophila in which ectopic expression of DREF was targeted to the eye imaginal disc cells. Overexpression of DREF in eye imaginal disc cells enhanced the promoter activity of dE2F. The obtained results indicate that the DRE/DREF system activates transcription of the dE2F gene. PMID- 9748284 TI - The N-terminal methionine is a major determinant of the DNA binding specificity of MEF-2C. AB - Members of the MEF-2 family of transcriptional regulators positively modulate the activity of basic helix-loop-helix proteins in both myogenic and neurogenic cell lineages. Previous work had shown that MEF-2C(2-117), a protein fragment comprising the dimerization and DNA-binding domains of MEF-2C but lacking the N terminal methionine, bound to AT-rich DNA sequences with high affinity. MEF-2C(2 117) did not discriminate between different AT-rich sequences. We now report the in vitro DNA binding properties of a MEF-2C fragment containing the N-terminal methionine. Measurements of the apparent dissociation constants of the complexes of GG-MEF-2C(1-117) revealed that different AT-rich sequences are bound with different affinities; in particular MEF site containing DNA (CTATAAATAG) is bound preferentially to DNA containing a SRF site (CATAAATG). Strikingly, when the shorter AT run consisted of six alternating thymines and adenines, almost wild type affinity was observed. Irrespective of the particular DNA sequence, all circular dichroism spectra of the DNA complexes of GG-MEF-2C(1-117) were superimposable and characterized by an identical maximal ellipticity at 269.5 nm, suggesting similar DNA conformations. Bending analysis by circular permutation assay revealed that on complex formation MEF-2C(2-117) induced cognate DNA to bend by 49 degrees, while heterologous DNA remained unbent. In the presence of the N-terminal methionine, however, all DNA sequences were bent by 70 degrees. The above results suggest an important function for the N-terminal methionine in properly orientating MEF-2C on the DNA. PMID- 9748286 TI - Functional map of a placenta-specific enhancer of the human leukemia inhibitory factor receptor gene. AB - We recently reported a placenta-specific enhancer in the human leukemia inhibitory factor receptor (LIFR) gene and now show detailed characterization of the 226-base pair enhancer (-4625/-4400 nucleotides). Four of twenty-two mutants in linker analysis showed reduced promoter activities to 45, 30, 10, and 10%, respectively. Specific binding of region A (-4617/-4602) with nuclear extract was competed by a known Oct-1 oligo and supershifted by Oct-1 antibody. Specific binding of region B (-4549/-4535) was competed by a GATA oligo, but could not be supershifted by four GATA antibodies. Nevertheless, mutagenesis showed that critical bases in region B were identical to the GATA core motif, indicating that region B may bind to a novel GATA family transcription factor. The other two adjacent regions designated as region C (-4464/-4445) showed no known consensus binding sites, and their specific placental JEG-3 nuclear extract binding was not evident in nonplacental nuclear extracts and was not competed by a trophoblast specific element (TSE), indicating that region C is a novel placenta-specific element (PSE, CATTTCCTGAACTAGTTTTT). Footprinting localized the binding boundary of PSE-binding protein (PSEB), and three Gs were found to be important for specific PSE binding. UV cross-linking showed that PSEB had a molecular mass of approximately 160 kDa, substituting the PSE with two previously reported placenta elements TSE or chorionic somatomammotropin enhancer factor 1 (CSEF-1) motifs resulted in markedly different promoter activities, indicating that PSEB is indeed different from TSE binding protein or CSEF-1. These results are the first demonstration that a novel PSE is the major element for placenta-specific enhancer activity in human LIFR gene. PMID- 9748285 TI - Stimulation of p53-mediated transcriptional activation by the p53-binding proteins, 53BP1 and 53BP2. AB - p53 is a tumor suppressor protein that controls cell proliferation by regulating the expression of growth control genes. In a previous study, we identified two proteins, 53BP1 and 53BP2, that are able to bind to wild type but not to mutant p53 via the DNA-binding domain of p53. We isolated cDNAs expressing a full-length human 53BP1 clone, which predicts a protein of 1972 residues that can be detected in the H358 human lung carcinoma cell line. The 53BP1 and 53BP2 genes were mapped to chromosomes 15q15-21 and 1q41-42, respectively. Immunofluorescence studies showed three types of staining patterns for 53BP1 as follows: both cytoplasmic and nuclear, homogeneous nuclear, and a nuclear dot pattern. In contrast, 53BP2 localized exclusively to the cytoplasm, and this pattern did not change upon coexpression of wild type p53. Although our previous study revealed that p53 is not able to bind simultaneously to either 53BP1 or 53BP2 and to DNA carrying a consensus binding site, both 53BP1 and 53BP2 enhanced p53-mediated transcriptional activation and induced the expression of a p53-dependent protein, suggesting that these proteins might function in signal transduction pathways to promote p53 activity. PMID- 9748287 TI - Cloning and disruption of caPLB1, a phospholipase B gene involved in the pathogenicity of Candida albicans. AB - The Candida albicans PLB1 gene was cloned using a polymerase chain reaction-based approach relying on degenerate oligonucleotide primers designed according to the amino acid sequences of two peptide fragments obtained from a purified candidal enzyme displaying phospholipase activity (Mirbod, F., Banno, Y., Ghannoum, M. A., Ibrahim, A. S., Nakashima, S., Yasuo, K., Cole, G. T., and Nozawa, Y. (1995) Biochim. Biophys. Acta 1257, 181-188). Sequence analysis of a 6.7-kilobase pair EcoRI-ClaI genomic clone revealed a single open reading frame of 1818 base pairs that predicts for a pre-protein of 605 residues. Comparison of the putative candidal phospholipase with those of other proteins in data base revealed significant homology to known fungal phospholipase Bs from Saccharomyces cerevisiae (45%), Penicillium notatum (42%), Torulaspora delbrueckii (48%), and Schizosaccharomyces pombe (38%). Thus, we have cloned the gene encoding a C. albicans phospholipase B homolog. This gene, designated caPLB1, was mapped to chromosome 6. Disruption experiments revealed that the caplb1 null mutant is viable and displays no obvious phenotype. However, the virulence of strains deleted for caPLB1, as assessed in a murine model for hematogenously disseminated candidiasis, was significantly attenuated compared with the isogenic wild-type parental strain. Although deletion of caPLB1 did not produce any detectable effects on candidal adherence to human endothelial or epithelial cells, the ability of the caplb1 null mutant to penetrate host cells was dramatically reduced. Thus, phospholipase B may well contribute to the pathogenicity of C. albicans by abetting the fungus in damaging and traversing host cell membranes, processes which likely increase the rapidity of disseminated infection. PMID- 9748288 TI - Hypoxia regulates beta-enolase and pyruvate kinase-M promoters by modulating Sp1/Sp3 binding to a conserved GC element. AB - The transcription rates of glycolytic enzyme genes are coordinately induced when cells are exposed to low oxygen tension. This effect has been described in many cell types and is not restricted to species or phyla. In mammalian cells, there are 11 distinct glycolytic enzymes, at least 9 of which are induced by hypoxia. Recent reports described a role for the hypoxia-inducible factor-1 (HIF-1) in the transcriptional activation of lactate dehydrogenase A, aldolase-A, phosphoglycerate kinase, and enolase-1 genes. It is not known whether the HIF-1 factor acts exclusively to regulate these genes during hypoxia, or how the other genes of the pathway are regulated. In this paper, we describe analyses of the muscle-specific pyruvate kinase-M and beta-enolase promoters that implicate additional mechanisms for the regulation of glycolytic enzyme gene transcription by hypoxia. Transient transcription of a reporter gene directed by either promoter was activated when transfected muscle cells were exposed to hypoxia. Neither of these promoters contain HIF-1 binding sites. Instead, the hypoxia response was localized to a conserved GC-rich element positioned immediately upstream of a GATAA site in the proximal promoter regions of both genes. The GC element was essential for both basal and hypoxia-induced expression and bound the transcription factors Sp1 and Sp3. Hypoxia caused the progressive depletion of Sp3 determined by DNA binding studies and Western analyses, whereas Sp1 protein levels remained unchanged. Overexpression of Sp3 repressed expression of beta enolase promoters. It is concluded that hypoxia activates these glycolytic enzyme gene promoters by down-regulating Sp3, thereby removing the associated transcriptional repression. PMID- 9748289 TI - DAD1 is required for the function and the structural integrity of the oligosaccharyltransferase complex. AB - Asparagine-linked glycosylation is a highly conserved protein modification reaction that occurs in all eukaryotic organisms. The oligosaccharyltransferase (OST), which has its active site exposed on the luminal face of the endoplasmic reticulum (ER), catalyzes the transfer of preassembled high mannose oligosaccharides onto certain asparagine residues of nascent polypeptides. The mammalian OST complex was initially thought to be composed of three transmembrane proteins, ribophorin I (RI), ribophorin II (RII), and OST48. Most recently, a small integral membrane protein of 12 kDa called DAD1 has been identified as an additional member of the mammalian OST complex. A point mutation in the DAD1 gene is responsible for the temperature-sensitive phenotype of a baby hamster kidney derived cell line (tsBN7) that undergoes apoptosis at the non-permissive temperature. Furthermore, the mutant protein DAD1 is not detectable in tsBN7 cells 6 h after shifting the cells to the non-permissive temperature. This temperature-sensitive cell line offered unique opportunities to study the effects caused by the loss of one OST subunit on the other three subunits and also on N linked glycosylation. Western blot analysis of cell lysates showed that after 6 h at the non-permissive temperature, steady-state levels of the ribophorins were reduced by about 50%, and OST48 was barely detectable. On the other hand, steady state levels of other components of the rough ER, such as the alpha-subunits of the TRAP (translocon-associated membrane protein) and the Sec61 complex, which are components of the translocation apparatus, are not affected by the instability of the OST subunits. Furthermore, N-glycosylation of the ribophorins was seriously affected 6 h after shifting the cells to the non-permissive temperature, and after 12 h they were synthesized only in the non-glycosylated form. As may be expected, this defect in the OST complex at the non-permissive temperature caused also the underglycosylation of a secretory glycoprotein. We concluded that degradation of DAD1 at the non-permissive temperature not only affects the stability of OST48 and the ribophorins but also results in the functional inactivation of the OST complex. PMID- 9748290 TI - The active streptococcal hyaluronan synthases (HASs) contain a single HAS monomer and multiple cardiolipin molecules. AB - The functional sizes of the two streptococcal hyaluronan synthases (HASs) were determined by radiation inactivation analysis of isolated membranes. The native enzymes in membranes from Group A Streptococcus pyogenes HAS and Group C Streptococcus equisimilis HAS were compared with the recombinant proteins expressed in Escherichia coli membranes. Based on their amino acid sequences, the masses of these four proteins as monomers are approximately 48 kDa. In all cases, loss of enzyme activity was a simple single exponential function with increasing radiation dose. The functional sizes calculated from these data were identical for the four HASs at approximately 64 kDa. In contrast, the sizes of the proteins estimated by the loss of antibody reactivity on Western blots were essentially identical at 41 kDa for the four HAS species, consistently lower than the functional size by approximately 23 kDa. Matrix-assisted laser desorption time of flight mass spectrometry analysis of purified S. pyogenes HAS-H6 and S. equisimilis HAS-H6 gave masses that differed by <0.07% from the predicted monomer sizes, which confirms that neither protein is posttranslationally modified or covalently attached to another protein. Ongoing studies indicate that the purified HAS enzymes require cardiolipin (CL) for maximal activity and stability. When irradiated membranes were detergent solubilized and the extracts were incubated with exogenous CL, the residual level of HAS activity increased. Consequently, the calculated functional size decreased by approximately 23 kDa to the expected size of the HAS monomer. The approximately 23-kDa larger size of the functional HAS enzyme, compared with the HAS monomer, is due, therefore, to CL molecules. We propose that the active streptococcal HA synthases are monomers in complex with approximately 16 CL molecules. PMID- 9748291 TI - Terminator-specific recycling of a B1-Alu transcription complex by RNA polymerase III is mediated by the RNA terminus-binding protein La. AB - Efficient synthesis of many small abundant RNAs is achieved by the proficient recycling of RNA polymerase (pol) III and stable transcription complexes. Cellular Alu and related retroposons represent unusual pol III genes that are normally repressed but are activated by viral infection and other conditions. The core sequences of these elements contain pol III promoters but must rely on fortuitous downstream oligo(dT) tracts for terminator function. We show that a B1 Alu gene differs markedly from a classical pol III gene (tRNAiMet) in terminator sequence requirements. B1-Alu genes that differ only in terminator sequence context direct differential RNA 3' end formation. These genes are assembled into stable transcription complexes but differ in their ability to be recycled in the presence of the La transcription termination factor. La binds to the nascent RNA 3' UUUOH end motif that is generated by transcriptional termination within the pol III termination signal, oligo(dT). We found that the recycling efficiency of the B1-Alu genes is correlated with the ability of La to access the 3' end of the nascent transcript and protect it from 3'-5' exonucleolytic processing. These results illuminate a relationship between RNA 3' end formation and transcription termination, and La-mediated reinitiation by pol III. PMID- 9748292 TI - Thiazolidinediones inhibit lipoprotein lipase activity in adipocytes. AB - The thiazolidinediones troglitazone and BRL 49653 improve insulin sensitivity in humans and animals with insulin resistance. Adipose tissue lipoprotein lipase is an insulin-sensitive enzyme. We examined the effects of thiazolidinediones on lipoprotein lipase expression in adipocytes. When added to 3T3-F442A, 3T3-L1, and rat adipocytes in culture, troglitazone and BRL 49653 inhibited lipoprotein lipase activity. This inhibition was observed at concentrations as low as 0.1 microM and within 2 h after addition of the drug. Lipoprotein lipase activity was inhibited in differentiated adipocytes as well as the differentiating cells. Despite this decrease in enzyme activity, these drugs increased mRNA levels in undifferentiated 3T3-F442A and 3T3-L1 cells and had no effect on mRNA expression or synthesis of lipoprotein lipase in differentiated cells. Western blot analysis showed that these drugs did not affect the mass of the enzyme protein. Lipoprotein lipase activity in cultured Chinese hamster ovary cells was not inhibited by troglitazone. Glucose transport, biosynthesis of lipids from glucose or the biosynthesis of proteins were unaffected by thiazolidinediones in differentiated cells, whereas glucose transport and lipid biosynthesis were increased when these drugs were added during differentiation. These results show that troglitazone and BRL 49653 have a specific, post-translational inhibitory effect on lipoprotein lipase in adipocytes, yet they promote lipid accumulation and differentiation in preadipocytes. PMID- 9748293 TI - Myocyte enhancer factor-related B-MEF2 is developmentally expressed in B cells and regulates the immunoglobulin J chain promoter. AB - Immunoglobulin J chain gene expression is induced by the delivery of a lymphokine signal to antigen-activated B cells in a primary immune response. A major interleukin 2 (IL-2)-responsive region that contains two adjacent control elements (JA and JB) exists within the J chain promoter. Transcription factor PU.1 positively regulates J chain gene expression by binding to one of the control elements (JB) in the J chain promoter. In the present study we have determined that a myocyte enhancer factor 2 (MEF2)-related nuclear factor, named B-MEF2, positively regulates the J chain gene promoter activity via the second control element (JA). An in vitro translated MEF2 family member, MEF2C, was found to bind the JA site with identical properties as endogenously expressed B-MEF2 in B cell lines. Moreover, in vivo experiments showed that a dominant negative mutant of MEF2C blocked B-MEF2 regulation of the J chain promoter. Consistent with its role as positive regulator of J chain gene expression, B-MEF2 levels were enhanced in highly differentiated B cells. In addition, induction of an IL-2 responsive presecretor cell line BCL1 with IL-2 or IL-5 (which up-regulates J chain gene expression) resulted in an increased expression of B-MEF2. We conclude that a MEF2-related transcriptional factor, B-MEF2, acts as a stage-specific positive regulator of J chain gene expression in the B cell lineage. PMID- 9748294 TI - Cloning and characterization of the 5'-flanking region for the human topoisomerase III gene. AB - The human DNA topoisomerase III (hTOP3) gene encodes a topoisomerase homologous to the Escherichia coli DNA topoisomerase I subfamily. To understand the mechanisms responsible for regulating hTOP3 expression, we have cloned the 5' flanking region of the gene coding for the hTOP3 and analyzed its promoter activity. The presence of a single transcription initiation site was suggested by primer extension analysis. The hTOP3 gene promoter is moderately high in GC content and lacks a canonical TATA box, suggesting that hTOP3 promoter has overall similarity to promoters of a number of housekeeping genes. Examination of the promoter sequence indicated the presence of four Sp-1 consensus binding sequences and a putative initiator element surrounding the transcription initiation site. Transient expression of a luciferase reporter gene under the control of serially deleted 5'-flanking sequences revealed that the 52-base pair region from -326 to -275 upstream of the transcription initiation site includes a positive cis-acting element(s) for the efficient expression of hTOP3 gene. On the basis of gel mobility shift and supershift assays, we demonstrated that both YY1 and USF1 transcription factors can bind to the 52-base pair region. When HeLa cells were transiently transfected with a mutant construct which had disabled both YY1- and USF1-binding sites, the luciferase activity was greatly reduced, suggesting that these binding elements play a functional role in the basal activation of the hTOP3 promoter. Transfection studies with mutations that selectively impaired YY1 or USF1 binding suggested that both YY1 and USF1 function as activators in the hTOP3 promoter. PMID- 9748295 TI - Differential stimulation of cholesterol and unsaturated fatty acid biosynthesis in cells expressing individual nuclear sterol regulatory element-binding proteins. AB - Three sterol regulatory element-binding proteins (SREBP-1a, -1c, and -2) stimulate transcription of genes involved in synthesis and receptor-mediated uptake of cholesterol and fatty acids. Here, we explore the individual roles of each SREBP by preparing lines of Chinese hamster ovary (CHO) cells that express graded amounts of nuclear forms of each SREBP (designated nSREBPs) under control of a muristerone-inducible nuclear receptor system. The parental hamster cell line (M19 cells) lacks its own nSREBPs, owing to a deletion in the gene encoding the Site-2 protease, which releases nSREBPs from cell membranes. By varying the concentration of muristerone, we obtained graded expression of individual nSREBPs in the range that restored lipid synthesis to near physiologic levels. The results show that nSREBP-2 produces a higher ratio of synthesis of cholesterol over fatty acids than does nSREBP-1a. This is due in part to a selective ability of low levels of nSREBP-2, but not nSREBP-1a, to activate the promoter for squalene synthase. nSREBP-1a and -2 both activate transcription of the genes encoding stearoyl-CoA desaturase-1 and -2, thereby markedly enhancing the production of monounsaturated fatty acids. nSREBP-1c was inactive in stimulating any transcription at the concentrations achieved in these studies. The current data support the emerging view that the nSREBPs act in complementary ways to modulate the lipid composition of cell membranes. PMID- 9748296 TI - Insulin-like growth factor I stimulates tyrosine phosphorylation of p130(Cas), focal adhesion kinase, and paxillin. Role of phosphatidylinositol 3'-kinase and formation of a p130(Cas).Crk complex. AB - Addition of insulin growth factor-I (IGF-I) to quiescent Swiss 3T3 cells rapidly induced tyrosine phosphorylation of the p130Crk-associated substrate (p130(Cas)), a novel adaptor protein localized at focal adhesions. Half-maximal effect was obtained at 0. 6 nM. IGF-I also promoted the formation of a complex between p130(Cas) and c-Crk and elicited a parallel increase in the tyrosine phosphorylation of p125(Fak) and paxillin. IGF-I-induced p130(Cas), p125(Fak), and paxillin tyrosine phosphorylation could be dissociated from mitogen-activated protein kinase kinase, p70(S6K), and protein kinase C activation. In contrast, the structurally unrelated phosphatidylinositol 3-kinase inhibitors wortmannin and LY294002 markedly attenuated the increase in tyrosine phosphorylation of p130(Cas), p125(Fak), and paxillin induced by IGF-I. Cytochalasin D, which disrupts the network of actin microfilaments, completely prevented tyrosine phosphorylation of p130(Cas), p125(Fak), and paxillin and the formation of a p130(Cas). Crk complex in response to IGF-I. Thus, our results identified a phosphatidylinositol 3-kinase-dependent pathway that requires the integrity of the actin cytoskeleton to induce tyrosine phosphorylation of p130(Cas), p125(Fak), and paxillin in response to IGF-I and suggest that tyrosine phosphorylation of these focal adhesion proteins, together with the recruitment of c-Crk into a complex with p130(Cas), may play a novel role in IGF-I signal transduction. PMID- 9748297 TI - A high fat diet impairs stimulation of glucose transport in muscle. Functional evaluation of potential mechanisms. AB - A high fat diet causes resistance of skeletal muscle glucose transport to insulin and contractions. We tested the hypothesis that fat feeding causes a change in plasma membrane composition that interferes with functioning of glucose transporters and/or insulin receptors. Epitrochlearis muscles of rats fed a high (50% of calories) fat diet for 8 weeks showed approximately 50% decreases in insulin- and contraction-stimulated 3-O-methylglucose transport. Similar decreases in stimulated glucose transport activity occurred in muscles of wild type mice with 4 weeks of fat feeding. In contrast, GLUT1 overexpressing muscles of transgenic mice fed a high fat diet showed no decreases in their high rates of glucose transport, providing evidence against impaired glucose transporter function. Insulin-stimulated system A amino acid transport, insulin receptor (IR) tyrosine kinase activity, and insulin-stimulated IR and IRS-1 tyrosine phosphorylation were all normal in muscles of rats fed the high fat diet for 8 weeks. However, after 30 weeks on the high fat diet, there was a significant reduction in insulin-stimulated tyrosine phosphorylation in muscle. The increases in GLUT4 at the cell surface induced by insulin or muscle contractions, measured with the 3H-labeled 2-N-4-(1-azi-2,2, 2-trifluoroethyl)-benzoyl-1,3-bis-(D mannose-4-yloxy)-2-propyla min e photolabel, were 26-36% smaller in muscles of the 8-week high fat-fed rats as compared with control rats. Our findings provide evidence that (a) impairment of muscle glucose transport by 8 weeks of high fat feeding is not due to plasma membrane composition-related reductions in glucose transporter or insulin receptor function, (b) a defect in insulin receptor signaling is a late event, not a primary cause, of the muscle insulin resistance induced by fat feeding, and (c) impaired GLUT4 translocation to the cell surface plays a major role in the decrease in stimulated glucose transport. PMID- 9748298 TI - Proteoglycans mediate cationic liposome-DNA complex-based gene delivery in vitro and in vivo. AB - The factors controlling cationic liposome-DNA complex (CLDC)-based gene transfer in cells and in animals are poorly understood. We found that cell surface heparin/heparan sulfate-bearing proteoglycans mediate CLDC-based gene transfer and expression both in cultured cells and following intravenous gene delivery into animals. CLDC did not transfect Raji cells, which lack proteoglycans, but did efficiently transfect Raji cells stably transfected with the proteoglycan, syndecan-1. Fucoidan, heparin, or dextran sulfate, all of which are highly anionic polysaccharides, each blocked CLDC-mediated transfection both in cultured cells and following intravenous injection into mice, but had no effect on transfection by either recombinant adenovirus infection or electroporation. Intravenous pretreatment of mice with heparinases, which specifically cleave heparan sulfate molecules from cell surface proteoglycans, blocked intravenous, CLDC-mediated transfection in mice, confirming that proteoglycans mediate CLDC gene delivery in vivo. Modulation of proteoglycan expression may prove useful in controlling the efficiency of, as well as targeting the sites of, CLDC-based gene transfer in animals. PMID- 9748299 TI - A novel glutathione peroxidase in bovine eye. Sequence analysis, mRNA level, and translation. AB - Bovine ciliary body contains a selenium-independent glutathione peroxidase (GPX) with a molecular mass of about 100 kDa that is composed of four identical subunits and exhibits no glutathione S-transferase activity. In this study, we isolated cDNA clones and determined the nucleotide sequence to deduce the primary structure of the enzyme. The cDNA contained 672 base pairs encoding a polypeptide with an estimated molecular mass of 25,064 Da. Translation of bovine ciliary mRNA produced a protein which was immunologically indistinguishable from GPX and showed high enzyme activity. The encoded amino acid sequence of the protein was 95% identical with that of a human keratinocyte gene product expressed in response to keratinocyte growth factor. It also showed sequence identity to bacterial alkyl hydroperoxide reductases and thiol specific antioxidant enzymes. GPX mRNA level was highest in the ciliary body, followed by the retina and iris. In various rat organs, the level of GPX mRNA was highest in the lung, followed by the muscle, liver, eye, heart, testis, thymus, kidney, and spleen. A very low level of mRNA was detected in the brain. Enzyme-linked immunosorbent assay with an antibody raised against the NH2-terminal sequence of GPX detected GPX protein in all rat tissues examined. PMID- 9748300 TI - Competitive inhibition of choline phosphotransferase by geranylgeraniol and farnesol inhibits phosphatidylcholine synthesis and induces apoptosis in human lung adenocarcinoma A549 cells. AB - We have previously shown that, among various isoprenoids, farnesol and geranylgeraniol specifically induced actin fiber disorganization, growth inhibition, and apoptosis in human lung adenocarcinoma A549 cells (Miquel, K., Pradines, A., and Favre, G. (1996) Biochem. Biophys. Res. Commun. 225, 869-876). Here we demonstrate that isoprenoid-induced apoptosis was preceded by an arrest in G0/G1 phase. The isoprenoid effects were independent of protein prenylation and of mitogen-activated protein kinase activity. Moreover, geranylgeraniol and farnesol induced a rapid inhibition of phosphatidylcholine biosynthesis at the last step of the CDP-choline pathway controlled by choline phosphotransferase and not at the level of CTP:phosphocholine cytidylyltransferase, the key enzyme of the pathway. Inhibition of choline phosphotransferase was confirmed by in vitro assays on microsomal fractions, which clearly showed that the isoprenoids acted by competitive inhibition with the diacylglycerol binding. Exogenous phosphatidylcholine addition prevented all the biological effects of the isoprenoids, including actin fiber disorganization and apoptosis, suggesting that inhibition of phosphatidylcholine biosynthesis might be the primary event of the isoprenoid action. These data demonstrate the molecular mechanism of geranylgeraniol and farnesol effects and suggest that the mevalonate pathway, leading notably to prenylated proteins, might be linked to the control of cell proliferation through the regulation of phosphatidylcholine biosynthesis. PMID- 9748301 TI - An unusual high-Km hexokinase is expressed in the mhAT3F hepatoma cell line. AB - In most hepatoma cells, the high-Km GLUT2/glucokinase proteins are replaced by the ubiquitous low-Km GLUT1/hexokinase type I proteins. In the mhAT3F hepatoma cells, the stimulatory effect of glucose on gene expression and glycogen accumulation was not maximal at 5 mmol/liter glucose. This response to high glucose is observed in mhAT3F cells, where GLUT2 was expressed, but not glucokinase (assessed by Northern blotting and reverse transcription-polymerase chain reaction). A low-Km hexokinase activity (19.6 +/- 3.8 milliunits/mg of protein) was present, but a high-Km (40 mmol/liter) hexokinase activity (13.9 +/- 2.5 milliunits/mg) was also detected in mhAT3F cells. The high-Km hexokinase activity was dependent on both ATP (or PPi) and glucose in the assay and was recovered in a 10-50-kDa fraction after filtration. A 30-kDa protein was detected using an anti-glucokinase antibody and localized by confocal microscopy at the same sites as glucokinase in hepatocytes. In FAO cells, the high-Km hexokinase activity and 30-kDa protein were not found. We conclude that a high-Km hexokinase activity is present in mhAT3F cells. This might explain why the effects of glucose on gene expression were not maximal at a glucose concentration of 5 mmol/liter. A 30-kDa protein identified using an anti-glucokinase antibody may be responsible for this activity present in mhAT3F cells. PMID- 9748302 TI - Leptin action in intestinal cells. AB - The adipocyte hormone leptin activates signal transducer and activator of transcription 3 (STAT3) in the hypothalamus, mediating increased satiety and increased energy expenditure. To date, leptin-mediated activation of the STAT pathway in vivo has not been established in tissues other than hypothalamus. We now describe leptin receptor expression and in vivo signaling in discrete regions of the mouse gastrointestinal tract. Expression of the functional isoform leptin receptor (OB-Rb) is restricted to the jejunum and is readily detected by RT-PCR in isolated enterocytes from this site. Intravenous injection of leptin rapidly induced nuclear STAT5 DNA binding activity in jejunum of +/+ and obese (ob/ob) mice but had no effect in the diabetic (db/db) mouse that lacks the OB-Rb isoform. In addition, an induction of the immediate-early gene c-fos is observed in jejunum in vivo. Leptin-mediated induction of a number of immediate-early genes and activation of STAT3 and STAT5 in a human model of small intestine epithelium, CACO-2 cells, corroborate this effect. Furthermore, intravenous leptin administration caused a significant 2-fold reduction in the apolipoprotein AIV transcript levels in jejunum 90 min after a fat load. Our results suggest that the epithelium of jejunum is a direct target of leptin action, and this activity is dependent on the presence of OB-Rb. Lack of leptin or resistance to leptin action in this site may contribute to obesity and its related syndromes by directly affecting lipid handling. PMID- 9748303 TI - The carboxyl terminus of Pneumocystis carinii glycoprotein A encodes a functional glycosylphosphatidylinositol signal sequence. AB - Pneumocystis carinii pneumonia is a hallmark disease associated with AIDS. An abundant glycoprotein, termed gpA, on the surface of P. carinii is considered an important factor in host-parasite interactions. The primary structure of ferret P. carinii gpA contains a carboxyl-terminal sequence characteristic of a signal for glycosylphosphatidylinositol (GPI) anchors. Here we report the capacity for this gpA carboxyl sequence to direct attachment of a secreted protein, human growth hormone (hGH), to the membranes of COS cells. A control fusion protein (hGHDAF37) was obtained which, under the direction of the GPI signal from decay accelerating factor, directs hGH cell surface expression. A construct (phGH2 1A30) was created similar to hGHDAF37 by fusing hGH to the putative GPI signal sequence encoded in the terminal 30 residues from a ferret P. carinii gpA cDNA clone. By indirect immunofluorescent staining, hGH was detected on the surface of COS cells transfected with phGH2-1A30; this surface location was confirmed by confocal laser cytometry. Metabolic labeling with [3H]ethanolamine and subsequent immunopurification of hGH from cells transfected with phGH2-1A30 confirmed that a lipid moiety characteristic of a conventional GPI anchor was linked covalently to hGH, and cell surface hGH2-1A30 fusion protein was sensitive to enzymatic cleavage by phosphatidylinositol-phospholipase C. Furthermore, hGH2-1A30 recombinant protein cofractionated with 5'-nucleotidase, a classical GPI-anchored membrane marker. Together, these results indicate that the carboxyl-terminal residues of ferret P. carinii gpA constitute a biologically functional GPI consensus domain, thus providing a potential mechanism for antigenic variation of P. carinii gpA during P. carinii pneumonia. PMID- 9748304 TI - Subunit folding and assembly steps are interspersed during Shaker potassium channel biogenesis. AB - In the voltage-dependent Shaker K+ channel, distinct regions of the protein form the voltage sensor, contribute to the permeation pathway, and recognize compatible subunits for assembly. To investigate channel biogenesis, we disrupted the formation of these discrete functional domains with mutations, including an amino-terminal deletion, Delta97-196, which is likely to disrupt subunit oligomerization; D316K and K374E, which prevent proper folding of the voltage sensor; and E418K and C462K, which are likely to disrupt pore formation. We determined whether these mutant subunits undergo three previously identified assembly events as follows: 1) tetramerization of Shaker subunits, 2) assembly of Shaker (alpha) and cytoplasmic beta subunits, and 3) association of the amino and carboxyl termini of adjacent Shaker subunits. Delta97-196 subunits failed to establish any of these quaternary interactions. The Delta97-196 deletion also prevented formation of the pore. The other mutant subunits assembled into tetramers and associated with the beta subunit but did not establish proximity between the amino and carboxyl termini of adjacent subunits. The results indicate that oligomerization mediated by the amino terminus is required for subsequent pore formation and either precedes or is independent of folding of the voltage sensor. In contrast, the amino and carboxyl termini of adjacent subunits associate late during biogenesis. Because subunits with folding defects oligomerize, we conclude that Shaker channels need not assemble from pre-folded monomers. Furthermore, association with native subunits can weakly promote the proper folding of some mutant subunits, suggesting that steps of folding and assembly alternate during channel biogenesis. PMID- 9748305 TI - Synergistic activation of the N-methyl-D-aspartate receptor subunit 1 promoter by myocyte enhancer factor 2C and Sp1. AB - The N-methyl-D-aspartate (NMDA) subtype of glutamate receptor plays important roles in neuronal development, plasticity, and cell death. NMDA receptor subunit 1 (NR1) is an essential subunit of the NMDA receptor and is developmentally expressed in postnatal neurons of the central nervous system. Here we identify on the NR1 promoter a binding site for myocyte enhancer factor 2C (MEF2C), a developmentally expressed neuron/muscle transcription factor found in cerebrocortical neurons, and study its regulation of the NR1 gene. Co-expression of MEF2C and Sp1 cDNAs in primary neurons or cell lines synergistically activates the NR1 promoter. Disruption of the MEF2 site or the MEF2C DNA binding domain moderately reduces this synergism. Mutation of the Sp1 sites or the activation domains of Sp1 protein strongly reduces the synergism. Results of yeast two hybrid and co-immunoprecipitation experiments reveal a physical interaction between MEF2C and Sp1 proteins. The MEF2C DNA binding domain is sufficient for this interaction. Dominant-negative MEF2C interferes with expression of NR1 mRNA in neuronally differentiated P19 cells. Growth factors, including epidermal growth factor and basic fibroblast growth factor, can up-regulate NR1 promoter activity in stably transfected PC12 cells, even in the absence of the MEF2 site, but the Sp1 sites are necessary for this growth factor regulation, suggesting that Sp1 sites may mediate these effects. PMID- 9748306 TI - Transcriptional regulation of the generic promoter III of the rat prolactin receptor gene by C/EBPbeta and Sp1. AB - Three promoters are operative in the rat prolactin receptor gene as follows: promoter I (PI) and II (PII) are specific for the gonads and liver, respectively, and promoter III (PIII) is common to several tissues. To investigate the mechanisms controlling the activity of promoter III, its regulatory elements and transcription factors were characterized in gonadal and non-gonadal cells. The TATA-less PIII domain was localized to the region -437 to -179 (ATG +1) containing the 5'-flanking region and part of the non-coding first exon. Within the promoter domain, a functional CAAT-box/enhancer binding protein (C/EBP) ( 398) and an Sp1 element (-386), which bind C/EBPbeta and Sp1/Sp3, respectively, contribute individually to promoter activation in gonadal and non-gonadal cells. However, significant redundancy was demonstrated between these elements in non gonadal cells. Additionally, an element within the non-coding exon 1 (-338) is also required for promoter activity. Activation of PIII by the widely expressed Sp1 and C/EBPbeta factors explains its common utilization in multiple tissues. Moreover, whereas the rat and mouse PIII share similar structure and function, the mouse PI lacks the functional SF-1 element and hence is inactive. These findings indicate that promoter III is of central importance in prolactin receptor gene transcription across species. PMID- 9748307 TI - Intracellular maturation of the mouse metalloprotease disintegrin MDC15. AB - Metalloprotease disintegrins are a family of membrane-anchored glycoproteins that play a role in fertilization, myoblast fusion, neuronal development, and cleavage of the membrane-anchored cytokine tumor necrosis factor-alpha. Here, we report the cloning and cDNA sequencing of the mouse metalloprotease disintegrin MDC15 and an analysis of its processing in the secretory pathway. A notable difference between mMDC15 and its putative human orthologue (hMDC15, metargidin) is the presence of the peptide sequence TDDC instead of the RGDC found in the disintegrin domain of hMDC15. In a Western blot analysis the majority of mMDC15 was found to lack the pro-domain in all mouse tissues examined. Pulse-chase experiments in transiently transfected COS-7 cells suggest that mMDC15 is processed by a pro-protein convertase in a late Golgi compartment, since (i) addition of brefeldin A or monensin blocks pro-domain removal, (ii) all detectable processed mMDC15 is endoglycosidase H -resistant, and (iii) a recombinant soluble form of the trans-Golgi network pro-protein convertase furin can mimic mMDC15 processing in vitro. Cell-surface trypsinization revealed that more than half of mature mMDC15 is intracellular. Immunolocalization provided evidence for a strong perinuclear accumulation in a region resembling the trans Golgi network and/or endosomal compartments. This study provides the first characterization of the intracellular processing of a metalloprotease disintegrin, and highlights the potential role of pro-protein convertases in removal of the inhibitory pro-domain. These results further suggest possible intracellular functions for mMDC15, such as in protein maturation, in addition to a potential role in cell-surface proteolysis or cell adhesion. PMID- 9748308 TI - Purification of soluble cytochrome b5 as a component of the reductive activation of porcine methionine synthase. AB - In mammals, methionine synthase plays a central role in the detoxification of the rogue metabolite homocysteine. It catalyzes a transmethylation reaction in which a methyl group is transferred from methyltetrahydrofolate to homocysteine to generate tetrahydrofolate and methionine. The vitamin B12 cofactor cobalamin plays a direct role in this reaction by alternately accepting and donating the methyl group that is in transit from one substrate (methyltetrahydrofolate) to another (homocysteine). The reactivity of the cofactor intermediate cob(I)alamin renders the enzyme susceptible to oxidative damage. The oxidized enzyme may be returned to the catalytic turnover cycle via a reductive methylation reaction that requires S-adenosylmethionine as a methyl group donor, and a source of electrons. In this study, we have characterized an NADPH-dependent pathway for the reductive activation of porcine methionine synthase. Two proteins are required for the transfer of electrons from NADPH, one of which is microsomal and the other cytoplasmic. The cytoplasmic protein has been purified to homogeneity and is soluble cytochrome b5. It supports methionine synthase activity in the presence of NADPH and the microsomal component in a saturable manner. In addition, purified microsomal cytochrome P450 reductase and soluble cytochrome b5 reconstitute the activity of the porcine methionine synthase. Identification of soluble cytochrome b5 as a member of the reductive activation system for methionine synthase describes a function for this protein in non-erythrocyte cells. In erythrocytes, soluble cytochrome b5 functions in methemoglobin reduction. In addition, it identifies an additional locus in which genetic polymorphisms may play a role in the etiology of hyperhomocysteinemia, which is correlated with cardiovascular diseases. PMID- 9748309 TI - Functional and structural properties of the mitochondrial outer membrane receptor Tom20. AB - Tom20 is an outer mitochondrial membrane protein that functions as a component of the import receptor complex for cytoplasmically synthesized mitochondrial precursor proteins. The human homologue, hTom20, consists of an N-terminal membrane anchor region predicted between aa5-25 and a soluble cytosolic domain from aa30 to 145. To analyze the properties of hTom20, we have expressed several truncations of the cytosolic domain as fusion proteins with glutathione S transferase. Our studies reveal that the cytosolic region of hTom20 is a monomeric protein in solution containing two domains which are involved in different functions of the receptor. The N-terminal region is involved in membrane binding (aa30-60) and recognition of the cleavable matrix targeting signals (aa50-90). In addition, we have demonstrated that the receptor recognizes the alpha-helical state of the matrix targeting signal. The dissociation constant for this interaction in the presence of a detergent which induces this secondary structure is 0.6 microM, one-fifth the value in the absence of detergent. In aqueous solution, the region between aa30 and 60 is loosely folded and stabilized against proteolytic cleavage by interaction with detergents or a matrix targeting signal. Our work further shows that the remainder of the cytosolic domain of hTom20, aa60-145, is a compactly folded globular domain containing a region (aa90 145) that is critical for the recognition of proteins bearing internal signal sequences such as the uncoupling protein and porin. PMID- 9748310 TI - Characterization of the N-terminal targeting signal binding domain of the mitochondrial outer membrane receptor, Tom20. AB - hTom20 is an outer mitochondrial membrane receptor involved in protein translocation. The cytosolic domain (aa30-145) and selected truncated versions of this domain were overexpressed and purified to study the structure-function relationship of this protein. Our studies reveal that the secondary structure of the cytosolic domain is very resistant to unfolding by guanidine-HCl and urea and is stabilized mainly by hydrophobic interactions. However, the tertiary structure of the N-terminal targeting signal binding domain (aa30-90) is more flexible. The first 30 amino acids of the cytosolic domain (aa30-60) are involved in recognizing N-terminal targeting signals and in stabilizing the cytosolic domain on the lipid surface. Moreover, we show that specifically aa30-48 interact with the membrane surface; a construct containing aa48-145 will only bind to the membrane surface in the presence of an N-terminal targeting signal peptide. The C terminal region of hTom20 (aa141-145) interacts with the N-terminal region of hTom20, helping to stabilize the proper conformation of the N-terminal targeting signal binding domain. Finally, hTom20 interacts with the N-terminal targeting signal of preornithine carbamyl transferase fused to dihydrofolate reductase very weakly (Kd = 8 microM), as would be expected if this interaction was the first in a series orchestrated by the import receptor complex to draw the targeted protein into the mitochondrion. PMID- 9748311 TI - Proton selective substate of the mitochondrial permeability transition pore: regulation by the redox state of the electron transport chain. AB - The permeability transition pore of rat liver mitochondria can be closed by chelating free Ca2+, with respect to the passage of large molecules such as mannitol and sucrose. However, an apparent H+-conducting substate remains open under these conditions, as indicated by the persistence of maximal O2 consumption rates and by the failure to recover a membrane potential. Agents which favor a closed pore, such as cyclosporin A, ADP, Mg2+, or bovine serum albumin, do not close the H+-conducting substate, but it closes spontaneously when respiration becomes limited by the availability of O2. Closure provoked by an O2 limitation requires free Mg2+ in the sub-micromolar concentration range and becomes less efficient with increasing time spent in the presence of free Ca2+. The H+ conducting substate is apparently regulated by the redox status of the electron transport chain, with a reduced form favoring closure. A physical association (or equivalence) between the pore and one of the respiratory chain complexes is supported. These characteristics suggest that the transition is irreversible in vivo, if it involves a small fraction of total mitochondria, and would lead to their elimination and/or replacement by the cell. The implications of this proposal are considered, as they relate to a possible role for the transition in cellular apoptosis and the elimination of mitochondria containing mutated DNA. PMID- 9748312 TI - A conformational change of the photoactive bacteriopheophytin in reaction centers from Rhodobacter sphaeroides. AB - It is demonstrated by ENDOR and Special TRIPLE spectroscopy that two distinct radical anion states of the intermediate electron acceptor (I), a bacteriopheophytin, can be freeze-trapped in isolated photosynthetic reaction centers of Rhodobacter sphaeroides. The formation of these states depends on the illumination time prior to freezing and the temperature. The first state, I1.-, is metastable and relaxes irreversibly at T approximately 160 K to the second state, I2.-. Experiments on quinone depleted as well as mutant reaction centers help to exclude the possibility that other cofactors besides the bacteriopheophytin in the A-branch, PhiA, are reduced during the trapping procedure. In particular, two mutants are investigated, in which the hydrogen bonds to PhiA that exist in the wild type are removed. These mutants are EL(L104), in which Glu at position L104 near the 13(1)-keto group of PhiA is replaced by Leu, and WF(L100), in which Trp at position L100 near the 13(2) methyl ester of PhiA is replaced by Phe. Both mutations have characteristic effects on both I.- states. In addition, the replacement of Thr at position M133 near the 13(1)-keto group of the inactive bacteriopheophytin and of Gly at position M203 near the 13(1)-keto group of the accessory bacteriochlorophyll in the A-branch by Asp causes no changes of the electronic structure of I.-. The two I.- states are interpreted in terms of a reorientation of the 3-acetyl group of PhiA after reduction. Possible implications for the initial charge separation process are discussed. PMID- 9748313 TI - The primary structures of the low-redox potential diheme cytochromes c from the phototrophic bacteria Rhodobacter sphaeroides and Rhodobacter adriaticus reveal a new structural family of c-type cytochromes. AB - The complete amino acid sequence of the low-redox potential cytochrome c-551.5 from Rhodobacter sphaeroides was determined by automated Edman degradation combined with mass spectroscopy. There are 139 residues and two typical Cys-X-X Cys-His heme-binding sites. A homologous low-redox potential cytochrome was also sequenced from Rhodobacter adriaticus and was found to contain 126 residues. It is 53% identical to that of Rb. sphaeroides and has two internal deletions of one and five residues. The Rhodobacter diheme cytochromes are 21-24% identical to the translated open reading frame SLL1886 from Synechocystis sp. PCC6801. There are at least two deletions of five and eight residues in the 188-residue cyanobacterial protein. Each of the three cytochromes has more histidines than it needs to bind the two hemes, but conserved histidines located 23 residues after the first heme and 14-19 residues before the second heme are likely to be the sixth heme ligands. There is no evidence for gene doubling and no similarity to any other known cytochromes. The measured helix content of 24% is much less than normal for c-type cytochromes. These proteins thus appear to be representative of an entirely new class of c-type cytochromes. PMID- 9748314 TI - Conversion of mitochondrial cytochrome b5 into a species capable of performing the efficient coupled oxidation of heme. AB - Histidine-63, one of the heme axial ligands in outer mitochondrial membrane cytochrome b5 (OM cyt b5) has been replaced by a methionine. The H63M variant performs the efficient and regioselective coupled oxidation of heme in order to produce >90% of the alpha-isomer of verdoheme. The variant was characterized by electronic, EPR, and NMR spectroscopic studies which indicate that the ferric form is a high-spin species whose heme is coordinated by histidine-39 in the proximal site and likely by water in the distal site. The coordination of methionine to the ferric heme was ruled out on the basis of NMR spectroscopic studies. Addition of imidazole to a solution of the ferric variant results in the formation of a species axially coordinated by imidazole and histidine-63. The reduction potential of the variant was found to be +110 mV in the absence of exogenous imidazole and -92 mV in the presence of imidazole. These values compare well with the reduction potential of myoglobin (50 mV) and wild-type OM cyt b5 ( 102 mV), respectively, consistent with the axial ligation described above. The ferrous variant, on the other hand, is a low-spin species coordinated by histidine-39 and methionine-63. Carbon monoxide (CO) readily displaces Met-63 from its coordination site on the ferrous heme, whereas CO cannot completely displace Met-63 from its coordination site on verdoheme. Consequently, the mechanism of inhibition for the oxidation of verdoheme to iron-biliverdin in the H63M variant appears to be similar to that observed for the heme-heme oxygenase complex in the presence of CO. PMID- 9748315 TI - Direct voltammetric observation of redox driven changes in axial coordination and intramolecular rearrangement of the phenylalanine-82-histidine variant of yeast iso-1-cytochrome c. AB - Direct square-wave and cyclic voltammetric electrochemical examination of the yeast iso-1-cytochrome c Phe82His/Cys102Ser variant revealed the intricacies of redox driven changes in axial coordination, concomitant with intramolecular rearrangement. Electrochemical methods are ideally suited for such a redox study, since they provide a direct and quantitative visualization of specific dynamic events. For the iso-1-cytochrome c Phe82His/Cys102Ser variant, square-wave voltammetry showed that the primary species in the reduced state is the Met80 Fe2+-His18 coordination form, while in the oxidized state the His82-Fe3+-His18 form predominates. The addition or removal of an electron to the appropriate form of this variant serves as a switch to a new molecular form of the cytochrome. Using the 2 x 2 electrochemical mechanism, simulations were done for the cyclic voltammetry experiments at different scan rates. These, in turn, provided relative rate constants for the intramolecular rearrangement/ligand exchange and the equilibrium redox potentials of the participating coordination forms: kb,AC = 17 s-1 for Met80-Fe3+-His18 --> His82-Fe3+-His18 and kf,BD > 10 s-1 for His82 Fe2+-His18 --> Met80-Fe2+-His18; E0' = 247 mV for Met80-Fe3+/2+-His18 couple, E0' = 47 mV for His82-Fe3+/2+-His18 couple, and E0' = 176 mV for the cross-reaction couple, His82-Fe3+-His18 + e- --> Met80-Fe2+-His18. Thermodynamic parameters, including the entropy of reaction, DeltaS0'Rxn, were determined for the net reduction/rearrangement reaction, His82-Fe3+-His18 + e- --> Met80-Fe2+-His18, and compared to those for wild-type cytochrome, Met80-Fe3+-His18 + e- --> Met80-Fe2+ His18. For the Phe82His variant mixed redox couple, DeltaS0'Rxn = -80 J/mol.K compared to DeltaS0'Rxn = -52 J/mol.K for the wild-type cyt c couple without rearrangement. Comparison of these entropies indicates that the oxidized His82 Fe3+-His18 form is highly disordered. It is proposed that this high level of disorder facilitates rapid rearrangement to Met80-Fe2+-His18 upon reduction. PMID- 9748316 TI - The active site of the bacterial nitric oxide reductase is a dinuclear iron center. AB - A novel, improved method for purification of nitric oxide reductase (NOR) from membranes of Paracoccus denitrificans has been developed. The purified enzyme is a cytochrome bc complex which, according to protein chemical and hydrodynamic data, contains two subunits in a 1:1 stoichiometry. The purified NorBC complex binds 0.87 g of dodecyl maltoside/g of protein and forms a dimer in solution. Similarly, it is dimeric in two-dimensional crystals. Images of these crystals have been processed at 8 A resolution in projection to the membrane. The NorB subunit is homologous to the main catalytic subunit of cytochrome oxidase and is predicted to contain the active bimetallic center in which two NO molecules are turned over to N2O. Metal analysis and heme composition implies that it binds two B-type hemes and a nonheme iron but no copper. NorC is a membrane-anchored cytochrome c. Fourier transform infrared spectroscopy shows that carbon monoxide dissociates from the reduced heme in light and associates with another metal center which is distinct from the copper site of heme/copper oxidases. Electron paramagnetic resonance spectroscopy reveals that NO binds to the reduced enzyme under turnover conditions giving rise to signals near g = 2 and g = 4. The former represents a typical nitrosyl-ferroheme signal whereas the latter is a fingerprint of a nonheme iron/NO adduct. We conclude that the active site of NOR is a dinuclear iron center. PMID- 9748317 TI - Change in environment of the P1 side chain upon progression from the Michaelis complex to the covalent serpin-proteinase complex. AB - Serpins inhibit proteinases by forming a kinetically trapped intermediate during a suicide substrate inhibition reaction. To determine whether the kinetic trap involves a repositioning of the P1 side chain of the serpin following formation of the initial Michaelis complex, we used the tryptophan of a P1 M-->W variant of human alpha1-proteinase inhibitor as a fluorescent reporter group of the environment of the P1 side chain. The P1W variant was a valid model serpin and formed SDS-stable complexes with both trypsin and chymotrypsin with a stoichiometry of inhibition close to 1.0. Rates of inhibition of chymotrypsin for wild-type and variant alpha1-proteinase inhibitor differred only approximately 1.8-fold. Rates of inhibition of trypsin were, however, 25-fold lower for the variant than for the wild-type inhibitor. Steady-state fluorescence spectra showed a change in environment for the P1 side chain upon forming both covalent complex with trypsin or chymotrypsin and noncovalent complex with anhydrochymotrypsin. The P1 environments in the chymotrypsin and anhydrochymotrypsin complexes were, however, different. Fluorescence quenching studies confirmed the burial of the P1 side chain upon formation of both the noncovalent and covalent complexes, but were not able to discriminate between the solvent accessibility in these complexes. Stopped-flow fluorescence measurements resolved the covalent intramolecular reaction that led to covalent complex and showed that, during the course of the covalent reaction, the environment of the P1 side chain changed consistent with a repositioning relative to residues of the proteinase active site as part of formation of the trap. This repositioning is likely to be a crucial part of the trapping mechanism. PMID- 9748318 TI - Mutual stabilization of VL and VH in single-chain antibody fragments, investigated with mutants engineered for stability. AB - A set of six mutants of the levan binding single-chain Fv (scFv) fragment A48 (ABPC48), which have the identical light chain but differ gradually in the stability of the heavy chain, was generated. This was achieved by introducing one or both of the stabilizing mutations H-K66R and H-N52S into the VH domain of the A48 wild-type protein, which is naturally missing the conserved disulfide bridge in VH, and into the cysteine-restored variant A48cys scFv. The stabilizing effects of these two mutations in VH, which had been selected in the context of a disulfide-free derivative of this scFv fragment [Proba, K., et al. (1998) J. Mol. Biol. 275, 245-253], were found to be additive and transferable to the cysteine restored variant of the A48 scFv, thereby generating extremely stable VH domains. The equilibrium denaturation of these scFv fragments was compared with the corresponding isolated VL domain and two of the different isolated VH domains. In the scFv fragment, the VL domain was found to be stabilized by a more stable VH domain, and, conversely, the VH domain was stabilized by a more stable VL domain. A folding intermediate with nativelike VH and denatured VL was found at equilibrium, if VH was significantly more stable than VL. In all other cases, a cooperative unfolding of the scFv was observed. We explain this observation with different contributions of intrinsic domain stability and extrinsic stabilization provided by the partner domain in the single-chain antibodies. PMID- 9748319 TI - Combination of gene targeting and substrate trapping to identify substrates of protein tyrosine phosphatases using PTP-PEST as a model. AB - Identification of physiological substrates of protein tyrosine phosphatases is a key step in understanding the function of these enzymes. We have generated fibroblast cell lines having a gene-targeted PTP-PEST in order to identify potential substrates with the premise that specific substrates of this enzyme would exist in a hyperphosphorylated state. Analysis of the profile of the phosphotyrosine proteins in the PTP-PEST -/- cells revealed the presence of hyperphosphorylated proteins of 180, 130, and 97 kDa when compared to control cells. The p130 was identified as p130(Cas), and direct immunoprecipitates of p130(Cas) demonstrate that this protein is constitutively hyperphosphorylated in cells lacking PTP-PEST. In addition, p130(Cas) was also isolated by the substrate trapping mutant of PTP-PEST in the PTP-PEST -/- cell lysates. Interestingly, we have demonstrated for the first time that PTP-PEST, through its first proline rich sequence 332PPKPPR337, interacts with other members of the p130(Cas) family (Hef1 and Sin) via their SH3 domain in vitro. This result suggests that Hef1 and Sin could also be potential substrates of PTP-PEST. In conclusion, we have combined genetic and biochemical strategies to allow the identification of PTP PEST substrates. This experimental approach could potentially be used to identify substrates of other PTPases. Furthermore, the Cas-like molecules Hef1 and Sin associate via their SH3 domains with a proline-rich motif found on PTP-PEST, suggesting the possibility that PTP-PEST could be a general modulator of the Cas family of proteins. PMID- 9748320 TI - Elucidation of the structural basis for the slow reactivity of thrombin with plasminogen activator inhibitor-1. AB - Plasminogen activator inhibitor-1 (PAI-1) is a serine protease inhibitor of the serpin superfamily which rapidly inactivates tissue plasminogen activator (tPA), but reacts with thrombin at a much slower rate. Based on the previous mutagenesis studies and the X-ray crystal structure of the thrombin E192Q-bovine pancreatic trypsin inhibitor (BPTI) complex, the structural basis for the slow reactivity of thrombin with PAI-1 is investigated in this study. In the crystal structure of the thrombin E192Q-BPTI complex, the reactive site loop of BPTI is stabilized in a canonical conformation by several productive interactions (e.g., Glu39 of thrombin is ion-paired to the P5' Arg, and Gln192 is hydrogen-bonded to the P2 and P4 backbone carbonyls of BPTI). PAI-1 contains Glu residues at both the P4' and P5' positions, and previous mutagenesis studies suggest that these residues make productive interactions with Arg39 of tPA as well as with two other positively charged residues present on the 39-loop of the protease (chymotrypsin numbering). Glu39 and Glu192 of thrombin would be unable to make such productive interactions with PAI-1. Instead, their repulsive interactions with the similarly charged residues and/or the backbone carbonyls of the PAI-1 reactive site loop could restrict the reaction. To test this, the rate constants (k2) for the PAI-1 inactivation of wild-type, E39K, E39Q, E192Q, E192M, and E39K/E192Q thrombins were determined. The inactivation rates of E39K [k2 = (4.3 +/- 0.2) x 10(4) M-1 s 1] and E39Q [k2 = (1.0 +/- 0.1) x 10(4) M-1 s-1] were 50- and 12-fold faster than the inactivation of wild-type thrombin [k2 = (8.6 +/- 0. 5) x 10(2) M-1 s-1], respectively. Relative to thrombin, the PAI-1 inactivation rates were improved 31 fold for E192Q [k2 = (2.7 +/- 0. 5) x 10(4) M-1 s-1] and 5-fold for E192M [k2 = (4.3 +/- 0.8) x 10(3) M-1 s-1] thrombins. With the double mutant E39K/E192Q, the inactivation rate [k2 = (5.4 +/- 0.4) x 10(5) M-1 s-1] was improved 628-fold over wild-type thrombin. These results suggest that repulsive interactions and/or lack of productive electrostatic interactions between PAI-1 and Glu39 and Glu192 of thrombin are responsible for the slow reaction of thrombin with this serpin. PMID- 9748321 TI - Demonstration of exosite I-dependent interactions of thrombin with human factor V and factor Va involving the factor Va heavy chain: analysis by affinity chromatography employing a novel method for active-site-selective immobilization of serine proteinases. AB - In an essential step of blood coagulation, factor V is proteolytically processed by thrombin to generate the activated protein cofactor, factor Va, and to release the activation fragments E and C1. For the identification and characterization of sites of thrombin binding to factor V and its activation products, a new method was developed for immobilizing thrombin and other serine proteinases specifically (>/=92%) through their active sites and used in affinity chromatography studies of the interactions. Interactions of factor V with exosite I of thrombin were shown to regulate the factor V activation pathway from the 93% +/- 12% inhibition of the rate of activation correlated with specific binding of hirudin54-65 to this exosite. Chromatography of factor V on active-site-immobilized thrombin showed only a weak interaction, while the factor Va heterodimer bound specifically and with apparently higher affinity, in an interaction that was prevented by hirudin54-65. The heavy chain of subunit-dissociated factor Va bound to immobilized thrombin, while the light-chain subunit and fragment E had no detectable affinity. These results demonstrate a previously undescribed, exosite I-dependent interaction of thrombin with factor Va that occurs through the factor Va heavy chain. They support the further conclusion that similar exosite I dependent binding of thrombin to the heavy-chain region of factor V contributes to recognition of factor V as a specific thrombin substrate and thereby regulates proteolytic activation of the protein cofactor. PMID- 9748322 TI - Heparan sulfate proteoglycans control intracellular processing of bFGF in vascular smooth muscle cells. AB - Basic fibroblast growth factor (bFGF) is a potent mitogen for vascular smooth muscle cells (VSMC) and has been implicated in a number of vascular disorders. bFGF interacts with high-affinity receptors and heparan sulfate proteoglycans (HSPG) at the cell surface. HSPG have been demonstrated to enhance bFGF binding to its receptors, yet no known role for HSPG in modulating postbinding events has been identified. In the present study, we analyzed bFGF internalization, intracellular distribution, degradation, and stimulation of DNA synthesis within native and HSPG-deficient VSMC. HSPG-deficient VSMC were generated by treating cells with sodium chlorate to inhibit the sulfation of HSPG. We found that stimulation of DNA synthesis by bFGF in chlorate-treated VSMC was markedly reduced as compared with native cells, even at doses of bFGF where receptor binding was similar in the two conditions. This was not a general lack of mitogenic potential, as the addition of calf serum, or epidermal growth factor, stimulated DNA synthesis to a similar extent in native and chlorate-treated cells. Analysis of the accumulation of internalized bFGF within cytoplasmic and nuclear fractions of native and HSPG-deficient VSMC showed striking differences. At early time points (0-2 h), nearly identical amounts of bFGF were observed in the cytoplasmic fractions under both conditions, yet significant amounts of bFGF were only found in the nuclear fractions of native cells. At later time points (2 48 h), the amount of cytoplasmic bFGF was significantly greater in the native compared to HSPG-deficient cells, and nuclear deposition of bFGF began to reach similar levels under both conditions. Furthermore, the intracellular half-life of bFGF was dramatically prolonged in native compared to HSPG-deficient cells, in part, due to decreased bFGF degradation in native cells. Thus, HSPG appears to accelerate nuclear localization, increase cytoplasmic capacity, and inhibit intracellular degradation of bFGF in VSMC. Modulation of intracellular processing of bFGF by HSPG might control the biological activity of bFGF in VSMC. PMID- 9748323 TI - Activation of distinct transcription factors in neutrophils by bacterial LPS, interferon-gamma, and GM-CSF and the necessity to overcome the action of endogenous proteases. AB - Human neutrophils can be induced to actively transcribe a number of early response genes, in particular those encoding cytokines, chemokines, and the high affinity surface receptor for IgG, FcgammaRI. Although little is known to date about the regulation of gene transcription in neutrophils, several indications point to a role for distinct transcription factors, such as members of the NF kappaB and STAT families. In this study, we investigated whether these transcription factors become activated under stimulatory conditions which are known to induce gene transcription in neutrophils. Unexpectedly, we found that conventional procedures employed to prepare cellular extracts cause the release of proteolytic activities that are normally stored in intracellular granules, resulting in the degradation of various NF-kappaB/Rel and STAT proteins. To circumvent this problem, we developed an alternative procedure which allowed us to show that in neutrophils, LPS and TNFalpha induce a NF-kappaB DNA-binding activity which essentially consists of p50/RelA dimers, and that IFNgamma promotes the binding of STAT1 homodimers to the IFNgamma response region of the FcgammaRI promoter. Moreover, we report that neutrophil stimulation with GM-CSF results in the formation of a STAT5-containing DNA-binding activity. Collectively, the current findings open new perspectives about mechanisms that are likely to regulate gene transcription in neutrophils. In addition, the procedure described herein could prove useful in other cell types that express high levels of endogenous proteases. PMID- 9748324 TI - Spontaneous inactivation of human tryptase involves conformational changes consistent with conversion of the active site to a zymogen-like structure. AB - The conformational changes accompanying spontaneous inactivation and dextran sulfate (DS) mediated reactivation of the serine protease human tryptase were investigated by analysis of (i) intrinsic fluorescence, (ii) inhibitor binding, and (iii) catalytic efficiency. Spontaneous inactivation produced a marked decrease in fluorescence emission intensity that was reversed by the addition of DS. Fluorescence decreases at high (4.0 microM) and low (0.1 microM) tryptase concentrations were similar at early times and coincided with loss of enzymatic activity but deviated significantly from activity loss at later times by showing a difference in the extent of change. The fluorescence losses were best described by a two-step kinetic model in which the major decrease correlated to activity loss (t1/2 of 4.3 min in 0.2 M NaCl, pH 6.8, 30 degrees C) and was followed by a further decrease (t1/2 approximately 60 min) whose extent differed with tryptase concentration. The ability to bind the competitive inhibitor p-aminobenzamidine was reversibly lost upon spontaneous inactivation, providing evidence for conformational changes affecting the major substrate binding site (S1-pocket). Estimation of catalytic efficiency using an active site titrant showed that the specific activity of tryptase remained unchanged upon inactivation and reactivation. Return of enzymatic activity, intrinsic fluorescence, and the S1 pocket appeared to occur in the same time frame (t1/2 approximately 3 min). These studies indicate that spontaneous inactivation involves reversible changes which convert the active site to a nonfunctional state. The association of activity loss with an intrinsic fluorescence decrease and loss of the S1-pocket is consistent with the disruption of a critical ionic bond at the active site. Formation of this ionic bond is the basis of zymogen activation for the chymotrypsin family of serine proteases. PMID- 9748325 TI - Mechanism-based inactivation of cytochrome P450 2B1 by 8-methoxypsoralen and several other furanocoumarins. AB - Of several furanocoumarins [5-methoxypsoralen (5-MOP), 8-methoxypsoralen (8-MOP), 5-hydroxypsoralen (5-OH-P), 8-hydroxypsoralen (8-OH-P), 4',5'-dihydro-8-MOP (DH-8 MOP), and psoralen (P)] tested as mechanism-based inactivators (MBIs) of purified reconstituted cytochrome P450 (P450) 2B1, 8-MOP was found to be the most potent (KI, kinact, and partition ratio of 2.9 microM, 0.34 min-1, and 1.3, respectively). The inactivation was not prevented by reactive oxygen species scavengers or nucleophilic trapping agents and proceeded with a decrease in P450 spectral content. Liquid chromatography (LC) separation of the reconstituted enzyme mixture, followed by liquid scintillation counting, indicated that [14C]-8 MOP binding was specific to the apoprotein of P450 2B1 with a binding stoichiometry of 0.7:1. The major metabolites formed by P450 2B1 from the furanocoumarins that were MBIs were characterized by LC electrospray ionization tandem mass spectrometry (ESI-MS/MS) as dihydro diols. Results from H218O incorporation experiments supported initial oxidation of 8-MOP and P to an epoxide which can react with some nucleophilic active site residue and inactivate the enzyme or partition to a dihydro diol metabolite by hydrolytic ring opening. On the other hand, 5-MOP was converted to an epoxide or gamma-keto enal intermediate prior to inactivation or dihydro diol formation. Comparison of the ESI mass spectra of P450 2B1 and furanocoumarin exposed P450 2B1, indicated a mass difference consistent with the covalent addition of a furanoepoxide to P450 2B1. PMID- 9748326 TI - Superoxide produced in the heme pocket of the beta-chain of hemoglobin reacts with the beta-93 cysteine to produce a thiyl radical. AB - The role of the beta-93 cysteine residue in the hemoglobin autoxidation process has been delineated by electron paramagnetic resonance. At low temperatures (8 K) after incubation at 235 K, free radical signals were detected. An analysis of the free radical spectrum produced implies that, besides the superoxide radical expected to be formed during autoxidation, an isotropic free radical is produced with a giso of 2.0133. This g value is consistent with that expected for a sulfur radical. Blocking the beta-93 sulfhydryl group with N-ethylmaleimide was found to eliminate the formation of the isotropic radical, but not the superoxide. This finding confirms the assignment of the isotropic radical as a thiyl radical originating from the oxidation of the cysteine SH group. A kinetic analysis of the time course for the formation of both the superoxide and thiyl radicals is consistent with a reversible electron transfer process between superoxide in the heme pocket of the beta-chains and the cysteine residue. This reaction is expected to produce both a thiyl radical and a peroxide. Direct evidence for peroxide production comes from the detection of a transient Fe(III) heme peroxide complex. The significance of the electron transfer process producing a thiyl radical is discussed. It is shown that the formation of the thiyl radical decreases the rate of autoxidation for the beta-chain and reduces heme degradation attributed to the reaction of superoxide with the heme. The insights gained from these low-temperature studies are believed to be relevant to room temperature autoxidation. PMID- 9748327 TI - Tryptophan-containing mutant of human (group IIa) secreted phospholipase A2 has a dramatically increased ability to hydrolyze phosphatidylcholine vesicles and cell membranes. AB - Human nonpancreatic (group IIa) secreted phospholipase A2 (human sPLA2) is associated with a number of inflammatory disorders in which the extracellular concentrations of this enzyme can become highly elevated. It is probable that the enzyme normally acts as an acute-phase protein whose function is to facilitate the removal of infectious organisms or damaged host cells as part of the normal inflammatory response. The enzyme shows negligible activity with phosphatidylcholine (PC) vesicles and cell membranes, presumably reflecting the enzyme's lack of ability to bind productively to such condensed neutral interfaces. Mammalian pancreatic enzymes show modest activity with such interfaces and contain a unique tryptophan at position 3, which is part of the presumptive interfacial binding surface of these enzymes. Human sPLA2 does not contain tryptophan. The amphiphilic indole side chain of tryptophan is noted for its ability to penetrate the lipid interface of membranes, and tryptophan residues appear to be associated with the ability of lipases and phospholipases A2 to bind to and hydrolyze such interfaces. We have investigated in detail the properties of a V3W mutant of human sPLA2, which has a unique tryptophan on the interfacial binding surface of this enzyme. Although this enzyme shows a modest ( approximately 50%) reduction in activity when anionic substrates are used under standard assay conditions, the activity of the enzyme on phosphatidylcholine vesicles and cell membranes is dramatically increased compared with human sPLA2. This is particularly the case with small unilamellar vesicles of PC, where activity is enhanced over 250-fold compared to the almost zero activity expressed by human sPLA2. This enhanced activity is best explained by increased interfacial binding and activation of the V3W mutant and is not due to enhanced active-site binding and hydrolysis. The results highlight the important role that tryptophan residues can play in interfacial binding, particularly to condensed zwitterionic interfaces. The interfacial characteristics of the mutant human enzyme now resemble more closely the mammalian pancreatic enzymes that already have a tryptophan at position 3. PMID- 9748328 TI - Allosteric activation of Arabidopsis threonine synthase by S-adenosylmethionine. AB - Plant threonine synthase, in contrast to its bacterial counterpart, is strongly stimulated by S-adenosylmethionine via a noncovalent interaction [Giovanelli et al. (1984) Plant. Physiol. 76, 285-292]. The mechanism of activation remained, however, largely unknown. To further characterize this unusual role for S adenosylmethionine, the Arabidopsis thaliana threonine synthase was overexpressed in Escherichia coli, purified to homogeneity, and then used for kinetic and enzyme-bound pyridoxal 5'-phosphate fluorescence equilibrium-binding experiments. We observed that the activating effect of S-adenosylmethionine results from an 8 fold increase in the rate of catalysis and from a 25-fold decrease in the Km value for the O-phosphohomoserine substrate. The data can be well fitted by a kinetic model assuming binding of two S-adenosylmethionine molecules on the native enzyme. We suggest that the dramatic modifications of the enzyme kinetic properties originate most presumably from an allosteric and cooperative transition induced by S-adenosylmethionine. This transition occurs at a much faster rate in the presence of the substrate than in its absence. PMID- 9748329 TI - Structure-function relationships in side chain lactam cross-linked peptide models of a conserved N-terminal domain of apolipoprotein E. AB - Bioactive peptides have multiple conformations in solution but adopt well-defined conformations at lipid surfaces and in interactions with receptors. We have used side chain lactam cross-links to stabilize secondary structures in the following peptide models of a conserved N-terminal domain of apolipoprotein E (cross-link periodicity in parentheses): I, H2N-GQTLSEQVQEELLSSQVTQELRAG-COOH (none); III, [sequence; see text] (i to i + 3); IV,[sequence; see text] (i to i + 4); IVa, [sequence, see text] (i to i + 4) (lactams above the sequence, potential salt bridges below the sequence). We previously demonstrated [Luo et al. (1994) Biochemistry 33, 12367-12377; Braddock et al. (1996) Biochemistry 35, 13975 13984] that peptide III, containing lactam cross-links between the i and i + 3 side chains, enhances specific binding of LDL via a receptor other than the LDL receptor. Peptide III in solution consists of two short alpha helices connected by a non alpha helical segment. Here we examine the hypothesis that the domain modeled by peptide III is one antipode of a conformational switch. To model another antipode of the switch, we introduced two strategic modifications into peptide III to examine structure-function relationships in this domain: (1) the spacing of the lactam cross-links was changed (i to i + 4 in peptides IV and IVa) and (2) peptides IV and IVa contain the two alternative sequences at a site of a possible end-capping interaction in peptide III. The structure of peptide IV, determined by 2D-NMR, is alpha helical across its entire length. Despite the remarkable degree of structural order, peptide IV is biologically inactive. In contrast, peptides III and possibly IVa contain a central interruption of the alpha helix, which appears necessary for biological activity. These and other studies support the hypothesis that this domain is a conformational switch which, to the extent that it models apolipoprotein E itself, may modulate interactions between apo E and its various receptors. PMID- 9748330 TI - Kinetic mechanism of Escherichia coli asparagine synthetase B. AB - Escherichia coli asparagine synthetase B (AS-B) catalyzes the synthesis of asparagine from aspartate, glutamine, and ATP. A combination of kinetic, isotopic labeling, and stoichiometry studies have been performed to define the nature of nitrogen transfer mediated by AS-B. The results of initial rate studies were consistent with initial binding and hydrolysis of glutamine to glutamate plus enzyme-bound ammonia. The initial velocity results were equally consistent with initial binding of ATP and aspartate prior to glutamine binding. However, product inhibition studies were only consistent with the latter pathway. Moreover, isotope-trapping studies confirmed that the enzyme-ATP-aspartate complex was kinetically competent. Studies using 18O-labeled aspartate were consistent with formation of a beta-aspartyl-AMP intermediate, and stoichiometry studies revealed that 1 equiv of this intermediate formed on the enzyme in the absence of a nitrogen source. Taken together, our results are most consistent with initial formation of beta -aspartyl-AMP intermediate prior to glutamine binding. This sequence leaves open many possibilities for the chemical mechanism of nitrogen transfer. PMID- 9748331 TI - The reaction mechanism for CD38. A single intermediate is responsible for cyclization, hydrolysis, and base-exchange chemistries. AB - Human recombinant CD38 catalyzes the formation of both cyclic ADP-ribose and ADP ribose products from NAD+ and hydrolyzes cyclic ADP-ribose to ADP-ribose. The corresponding GDP products are formed from NGD+. The enzyme was characterized by substrate and inhibition kinetics, exchange studies, rapid-quench reactions, and stopped-flow-fluorescence spectroscopy to establish the reaction mechanism and energetics for individual steps. Noncyclizable substrates NMN+ and nicotinamide-7 deaza-hypoxanthine dinucleotide (7-deaza NHD+) were rapidly hydrolyzed by the enzyme. The kcat for NMN+ was 5-fold higher than that of NAD+ and has the greatest reported kcat of any substrate for CD38. 7-deaza-NHD+ was hydrolyzed at approximately one-third the rate of NHD+ but does not form a cyclic product. These results establish that a cyclic intermediate is not required for substrate hydrolysis. The ratio of methanolysis to hydrolysis for cADPR and NAD+ catalyzed by CD38 increases linearly with MeOH concentration. Both reactions produce predominantly the beta-methoxy riboside compound, with a relative nucleophilicity of MeOH to H2O of 11. These results indicate the existence of a stabilized cationic intermediate for all observed chemistries in the active site of CD38. The partitioning of this intermediate between cyclization, hydrolysis, and nicotinamide-exchange unites the mechanisms of CD38 chemistries. Steady-state and pre-steady-state parameters for the partition and exchange mechanisms allowed full characterization of the reaction coordinate. Stopped-flow methods indicate a burst of cGDPR formation followed by the steady-state reaction rate. A lag phase, which was NGD+ concentration dependent, was also observed. The burst size indicates that the dimeric enzyme has a single catalytic site formed by two subunits. Pre-steady-state quench experiments did not detect covalent intermediates. Nicotinamide hydrolysis of NGD+ precedes cyclization and the chemical quench decomposes the enzyme-bound species to a mixture of cyclic and hydrolysis products. The time dependence of this ratio indicated that nicotinamide bond-breakage occurs 4 times faster than the conversion of the intermediate to products. Product release is the overall rate-limiting step for enzyme reaction with NGD+. PMID- 9748332 TI - ATP/GTP hydrolysis is required for oxazole and thiazole biosynthesis in the peptide antibiotic microcin B17. AB - In the maturation of the Escherichia coli antibiotic Microcin B17, the product of the mcbA gene is modified posttranslationally by the multimeric Microcin synthetase complex (composed of McbB, C, and D) to cyclize four Cys and four Ser residues to four thiazoles and four oxazoles, respectively. The purified synthetase shows an absolute requirement for ATP or GTP in peptide substrate heterocyclization, with GTP one-third as effective as ATP in initial rate studies. The ATPase/GTPase activity of the synthetase complex is conditional in that ADP or GDP formation requires the presence of substrate; noncyclizable versions of McbA bind to synthetase, but do not induce the NTPase activity. The stoichiometry of ATP hydrolysis and heterocycle formation is 5:1 for a substrate that contains two potential sites of modification. However, at high substrate concentrations (>50Km) heterocycle formation is inhibited, while ATPase activity occurs undiminished, consistent with uncoupling of NTP hydrolysis and heterocycle formation at high substrate concentrations. Sequence homology reveals that the McbD subunit has motifs reminiscent of the Walker B box in ATP utilizing enzymes and of motifs found in small G protein GTPases. Mutagenesis of three aspartates to alanine in these motifs (D132, D147, and D199) reduced Microcin B17 production in vivo and heterocycle formation in vitro, suggesting that the 45 kDa McbD has a regulated ATPase/GTPase domain in its N-terminal region necessary for peptide heterocyclization. PMID- 9748333 TI - Role of the propeptide and gamma-glutamic acid domain of factor IX for in vitro carboxylation by the vitamin K-dependent carboxylase. AB - The vitamin K-dependent gamma-glutamyl carboxylase catalyzes the processive carboxylation of specific glutamates in a number of proteins related to blood coagulation and bone. To address the independent importance of the propeptide, gamma-carboxyglutamic acid (Gla) domain and elements beyond the Gla domain of factor IX in vitamin K-dependent carboxylation, we have examined the kinetics of carboxylation of peptides containing (1) propeptide and Gla domain, (2) the Gla domain alone, (3) uncarboxylated bone Gla protein, (4) propeptide followed by the entire uncarboxylated factor IX molecule, and (5) the factor IX propeptide followed by a non-Gla domain sequence. Our studies indicate that peptides with a covalently linked propeptide have Km values similar to the physiological substrate of the carboxylase. In contrast, the Gla domain of factor IX has a >/=230-fold higher Km for the carboxylase than the corresponding peptide with a covalently linked propeptide. This contrasts with bone Gla protein, another vitamin K-dependent protein, which appears not to require a covalently linked propeptide for high-affinity binding to the carboxylase. Analysis of the carboxylation products of a propeptide/non-Gla domain substrate indicate that it is carboxylated multiple times in a processive manner. These studies show that the perceived binding affinity of the carboxylase substrate and processivity is conferred by the propeptide without requiring the conserved Gla domain sequences and that factor IX and bone Gla protein may have distinct mechanisms of interacting with the carboxylase. PMID- 9748334 TI - Ligand-induced conformational transitions in Escherichia coli phosphofructokinase 2: evidence for an allosteric site for MgATP2-. AB - The binding of ligands to phosphofructokinase 2 (Pfk-2) from Escherichia coli induces changes in the fluorescence emission properties of its single tryptophan residue, Trp88, suggesting that upon binding the protein undergoes a conformational change. This fluorescence probe was used to determine the presence of an allosteric site for MgATP2- in the enzyme. Fructose 6-phosphate (fructose-6 P), the first substrate that binds to the enzyme with an ordered bi-bi mechanism, increases the fluorescence up to 30%. The saturation curve for this compound is hyperbolic with a Kd of 6 microM. The titration of Pfk-2 with MgATP2- causes a quenching of fluorescence of about 30% of its initial value, with a blue shift of 7 nm in the emission maximum. The response is cooperative with a Kd of 80 microM and a Hill coefficient of 2. The interaction of MgATP2- cannot take place at the active site in the absence of fructose-6-P, due to the ordered kinetic mechanism. Addition of compounds that bind to the catalytic site of Pfk-2, such as ATP4- or Mg-AMP-PNP, did not produce significant changes in the fluorescence spectrum of Trp88. However, in the absence of Mg2+, the addition of ATP4- to the enzyme fructose-6-P complex shows a hyperbolic increase of fluorescence of 8%. Acrylamide steady-state quenching experiments for different enzyme-ligand complexes of Pfk-2, indicate that the tryptophan in the enzyme-MgATP2- complex is exposed to a much smaller extent to the solvent than in the free enzyme or in the enzyme-fructose-6-P complex. The effect of the binding of fructose-6-P or MgATP2- on the polarization fluorescence of the emission of Trp88 in Pfk-2 indicates changes in the local mobility of the Trp88 in both enzyme complexes. The average lifetime for Trp88 in Pfk-2 was found to be unusually large, approximately 7.7 ns, and did not vary significantly with the ligation state of the enzyme, which demonstrates that the quenching or enhancement of fluorescence induced by the ligands is mainly due to the complex formation with Pfk-2. These results demonstrate the presence of an allosteric site for MgATP2- in Pfk-2 from E. coli, responsible for the inhibition of the enzyme activity by this ligand. PMID- 9748335 TI - Kinetic studies on the effect of yeast cofilin on yeast actin polymerization. AB - The effect of yeast cofilin on the kinetics of polymerization of yeast actin has been examined at 20 degrees C at both pH 8.0 and 6.6. In the absence of cofilin, the kinetic data may be described by a simple nucleation-elongation mechanism. Kinetic data in the presence of cofilin suggests a complex dependence on the cofilin concentration. At low cofilin-to-actin ratios, cofilin increases the rate of polymerization in a way best fit by assuming filament fragmentation. The apparent fragmentation rate constants increase with increasing cofilin concentration leveling off above a cofilin-to-actin ratio of 1:8 and are independent of pH. At higher cofilin-to-actin ratios, a nonpolymerizable cofilin G-actin complex forms resulting in a decreased rate of polymerization. The data from fluorescence photobleaching recovery experiments at low cofilin-to-actin ratios are consistent with the presence of severed filaments at both pH 8 and 6.6. However, at pH 8 and a cofilin-to-actin ratio of 1:16, about 40-50% of the total actin is present as G-actin after polymerization while at pH 6.6 little or no G-actin is present at the same cofilin-to-actin ratio. The results suggest some cooperativity with respect to cofilin binding to filamentous actin which may be pH dependent. PMID- 9748336 TI - A specific amino acid sequence at the head-rod junction is not critical for the phosphorylation-dependent regulation of smooth muscle myosin. AB - It has been suggested that the structure at the head-rod junction of smooth muscle myosin is important for the phosphorylation-mediated regulation of myosin motor activity. To investigate whether a specific amino acid sequence at the head rod junction is critical for the regulation, three smooth muscle myosin mutants in which the sequence at the N-terminal end of S2 is deleted to various extents were expressed in Sf9 cells; 28, 56, and 84 amino acid residues, respectively, at the position immediately C-terminal to the invariant proline (Pro849) were deleted, and the S1 domain was directly linked to the downstream sequence of the rod. The mutant myosins were expressed, purified, and biochemically characterized. All three myosin mutants showed a stable double-headed structure based upon electron microscopic observation. Both the actin-activated ATPase activity and the actin translocating activity of the mutants were completely regulated by the phosphorylation of the regulatory light chain. The actin sliding velocity of the three mutant myosins was the same as the wild-type recombinant myosin. These results indicate that a specific amino acid sequence at the head rod junction is not required for the regulation of smooth muscle myosin. The results also suggest that there is no functionally important interaction between the regulatory light chain and the heavy chain at the head-rod junction. PMID- 9748337 TI - A novel high-affinity inhibitor for inward-rectifier K+ channels. AB - Inward-rectifier K+ channels are a group of highly specialized K+ channels that accomplish a variety of important biological tasks. Inward-rectifier K+ channels differ from voltage-activated K+ channels not only functionally but also structurally. Each of the four subunits of the inward-rectifier K+ channels has only two instead of six transmembrane segments compared to the voltage-activated K+ channels. Thus far, there are no high-affinity ligands that directly target any inward-rectifier K+ channel. In the present study, we identified, purified, and synthesized a protein inhibitor of the inward-rectifier K+ channels. The inhibitor, called tertiapin, blocks a G-protein-gated channel (GIRK1/4) and the ROMK1 channel with nanomolar affinities, but a closely related channel, IRK1, is insensitive to tertiapin. Mutagenesis studies show that teritapin inhibits the channel by binding to the external end of the ion conduction pore. PMID- 9748338 TI - Steady-state and pre-steady-state kinetic analysis of 8-oxo-7,8-dihydroguanosine triphosphate incorporation and extension by replicative and repair DNA polymerases. AB - The kinetics of 8-oxo-7,8-dihydroguanosine triphosphate (8-oxo-dGTP) incorporation into DNA by Escherichia coli polymerases I exo- (KF-) and II exo- (Pol II-), HIV-1 RT reverse transcriptase (HIV-1 RT), and bacteriophage T7 exo- (T7(-)) were examined to determine the misincorporation potential for 8-oxo-dGTP and to investigate the role of base pairing symmetry in DNA polymerase fidelity. 8-Oxo-dGTP was found to be a poor substrate for the four polymerases, with insertion efficiencies >10(4)-fold lower than for dGTP incorporation. Insertion efficiencies of 8-oxo-dGTP were also consistently lower than for incorporation of dNTPs opposite template 8-oxo-G, previously studied in this laboratory. In steady state reactions, T7(-) had a high preference for 8-oxo-dGTP insertion opposite A (97%) and HIV-1 RT, KF-, and Pol II- preferred to insert 8-oxo-dGTP opposite C. Misinsertion frequencies for 8-oxo-dGTP also varied considerably from frequencies of misinsertion at template 8-oxo-G adducts for Pol II-, HIV-1 RT, and T7(-). Pre steady-state incorporation of 8-oxo-dGTP opposite C (but not opposite A) by HIV-1 RT, KF-, and Pol II- displayed biphasic curves, with rates of initial incorporation 2- to 11-fold lower than normal dGTP incorporation. Although extension past template 8-oxo-G adducts had previously been shown to occur preferentially for the mispair, extension past primer 8-oxo-G:template A or C pairs was variable. The low and comparable estimated Kd values for dGTP and 8-oxo dGTP binding to HIV-1 RT alone or HIV-1 RT.DNA complexes indicated that the initial binding was nonselective and had high affinity. The large difference (>3 orders of magnitude) in kinetic Kdapp values for 8-oxo-dGTP and dGTP binding to HIV-1 RT.DNA indicates that there are contributions to the kinetically determined Kdapp (such as conformational change and/or phosphodiester bond formation) which may be involved in the selection against 8-oxo-dGTP. The differences in binding (Kdapp), incorporation, and extension kinetics of 8-oxo-dGTP compared to normal dNTP incorporation at template 8-oxo-G adducts indicate that polymerase fidelity does not depend solely upon the overall geometry of Watson-Crick base pairs and reflects the asymmetry of the enzyme active site. PMID- 9748339 TI - Spectroscopic characterization of a DNA-binding domain, Z alpha, from the editing enzyme, dsRNA adenosine deaminase: evidence for left-handed Z-DNA in the Z alpha DNA complex. AB - Double-stranded RNA adenosine deaminase (ADAR1) is an ubiquitous enzyme in metazoa that edits pre-mRNA changing adenosine to inosine in regions of double stranded RNA. Zalpha, an N-terminal domain of human ADAR1 encompassing 76 amino acid residues, shows apparent specificity for the left-handed Z-DNA conformation adopted by alternating (dGdC) polymers modified by bromination or methylation, as well as for (dGdC)13 inserts present in supercoiled plasmids. Here, a combination of circular dichroism, fluorescence, and gel-retardation studies is utilized to characterize recombinant Zalpha peptide and to examine its interaction with DNA. Results from laser-Raman spectroscopy experiments provide direct evidence for the existence of Z-DNA in peptide-DNA complexes. PMID- 9748340 TI - Optimization of alternate-strand triple helix formation at the 5'CpG3' and 5'GpC3' junction steps. AB - Oligonucleotide-directed triple helix formation normally requires a long tract of oligopyrimidine.oligopurine sequence. This limitation can be partially overcome by alternate-strand triple helix (or switch triple helix) formation which enables recognition of alternating oligopurine/oligopyrimidine sequences. The present work is devoted to the optimization of switch triple helix formation at the 5'CpG3' and 5'GpC3' junction steps by combination of base triplets in Hoogsteen and in reverse Hoogsteen configurations. Rational design by molecular mechanics was first carried out to study the geometrical constraints at different junction steps and to propose a "switch code" which would optimize the interactions at junctions. These predictions were further checked and validated experimentally by gel retardation and DNase I footprinting assays. It was shown that the choice of an appropriate linker nucleotide in the switching third strand plays an important role in the interaction between oligonucleotides and alternating oligopurine/oligopyrimidine target sequences at different junctions: (i) the addition of a cytosine at the junction level in the oligonucleotide optimizes the crossover at the 5'CpG3' junction, whereas (ii) the best crossover at the 5'GpC3' junction step is achieved without any additional nucleotide. These results provide a useful guideline to extend double-stranded DNA sequence recognition by switch triple helix formation. PMID- 9748341 TI - Mechanism and utility of an RNA-cleaving DNA enzyme. AB - We previously reported the in vitro selection of a general-purpose RNA-cleaving DNA enzyme that exhibits a catalytic efficiency (kcat/KM) exceeding that of any other known nucleic acid enzyme [Santoro, S. W. and Joyce, G. F. (1997) Proc. Natl. Acad. Sci. U.S.A. 94, 4262-4266]. This enzyme contains approximately 30 deoxynucleotides and can cleave almost any RNA substrate under simulated physiological conditions, recognizing the substrate through two Watson-Crick binding domains. The kinetics of cleavage under conditions of varying pH, choice of divalent metal cofactor, and divalent metal concentration are consistent with a chemical mechanism involving metal-assisted deprotonation of a 2'-hydroxyl of the substrate, leading to substrate cleavage. Kinetic measurements reveal that the enzyme strongly prefers cleavage of the substrate over ligation of the two cleavage products and that the enzyme's catalytic efficiency is limited by the rate of substrate binding. The enzyme displays a high level of substrate specificity, discriminating against RNAs that contain a single base mismatch within either of the two substrate-recognition domains. With appropriate design of the substrate-recognition domains, the enzyme exhibits a potent combination of high substrate sequence specificity and selectivity, high catalytic efficiency, and rapid catalytic turnover. PMID- 9748342 TI - Structural constraints in the HIV-1 reverse transcriptase-primer/template complex for the initiation of DNA synthesis from primer tRNALys3. AB - The topography and functional implications of the complex formed in vitro between human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) and its primer tRNALys3 were studied in this work. On the basis of previous results showing the high affinity both of the native primer, tRNALys3, as well as that of mismatched short oligonucleotide primers for HIV-1 RT, we synthesized chimeric primers containing tRNALys3 linked to U and T residues of different lengths. We found that the affinity of the oligonucleotide primers for HIV-1 RT is dramatically increased when linked to primer tRNA. Our results also show that in the tRNA.RT complex, before annealing tRNALys3 to the retroviral RNA genome, the 3'-terminal nucleotide of tRNALys3 is positioned at a distance of one nucleotide unit away from the template in the active polymerization site of the enzyme. PMID- 9748343 TI - Structure of Salmonella typhimurium nrdF ribonucleotide reductase in its oxidized and reduced forms. AB - The first class Ib ribonucleotide reductase R2 structure, from Salmonella typhimurium, has been determined at 2.0 A resolution. The overall structure is similar to the Escherichia coli class Ia enzyme despite only 23% sequence identity. The most spectacular difference is the absence of the pleated sheet and adjacent parts present in the E. coli R2 structure; the heart-shaped structure loses its tip. From sequence comparisons, it appears that this feature is shared with all other class Ib enzymes and, in this respect, is more like the mammalian class Ia enzymes. Both the oxidized and reduced iron forms have been investigated. In the ferric iron center, both iron ions are octahedrally coordinated and bridged by one carboxylate and one oxide ion. The ferrous form has lost the bridging oxide ion but is bridged by two carboxylates. Accompanying the change in redox state, helix E changes its conformation from one covering the metal center in the oxidized form to a more open reduced form. A narrow channel is opened which may permit easier access of oxygen to the ferrous iron site and to efficiently generate the tyrosyl radical. PMID- 9748344 TI - 14N electron spin-echo envelope modulation of the S = 3/2 spin system of the Azotobacter vinelandii nitrogenase iron-molybdenum cofactor. AB - Wild-type nitrogenase MoFe protein shows a deep 14N electron spin-echo envelope modulation (ESEEM) arising from a nitrogen nucleus (N1) coupled to the S = 3/2 spin system of the FeMo-cofactor of the MoFe protein. A previous ESEEM study on altered MoFe proteins generated by substitutions at the alpha-195-histidine position suggested that alpha-195-histidine provides a hydrogen bond to the FeMo cofactor but is not the source of the 14N1 modulation [DeRose et al. (1995) Biochemistry 34, 2809-2814]. This study also raised the possibility of a correlation between ESEEM spectroscopic properties and the nitrogenase phenotype. We now report ESEEM studies on altered MoFe proteins with substitutions at residues alpha-96-arginine, alpha-359-arginine, and alpha-381-phenylalanine to (i) assign the first-shell hydrogen bonding as revealed by the 14N modulation; (ii) explore the mechanistic relevance of the ESEEM signatures to nitrogenase activity; and (iii) study microscopic changes within the polypeptide environment of the FeMo-cofactor. Present ESEEM data reveals that two kinds of 14N modulations are present in wild-type MoFe protein. A new 2-dimensional procedure for high-precision analysis of the ESEEM data of the MoFe proteins shows that the deep wild-type ESEEM modulation (denoted N1) has a hyperfine-coupling constant of Aiso = 1.05 MHz and nuclear quadrupole coupling parameters of e2qQ = 2.17 MHz, eta = 0.59; the other (denoted N2) has a smaller hyperfine coupling of Aiso = approximately 0.5 MHz and e2qQ = approximately 3.5 MHz, eta = approximately 0.4. The N2 ESEEM pattern is more obvious when unmasked by substitutions that result in the loss of the deep N1 modulation. Correlations of the ESEEM properties and catalytic activities of the altered MoFe proteins suggest that (i) the side chain of the alpha-359-arginine is the source of the deep ESEEM N1 modulation; (ii) one or both of the amide nitrogens of alpha-356-glycine/alpha-357-glycine are responsible for the weak N2 modulation; (iii) substitution of the nonpolar alpha 381-phenylalanine residue, as well as substitution of either the alpha-195 histidine or alpha-359-arginine residues, can eliminate the N1 interaction with FeMo-cofactor; and (iv) ESEEM can be used to detect slight reorientations of FeMo cofactor within its polypeptide pocket, although the mechanistic relevance of the loss or perturbation of the hydrogen-bonding interactions between FeMo-cofactor and polypeptide environment has not yet been established. PMID- 9748345 TI - Is a hydrophobic amino acid required to maintain the reactive V conformation of thiamin at the active center of thiamin diphosphate-requiring enzymes? Experimental and computational studies of isoleucine 415 of yeast pyruvate decarboxylase. AB - The residue I415 in pyruvate decarboxylase from Saccharomyces cerevisiae was substituted with a variety of uncharged side chains of varying steric requirements to test the hypothesis that this residue is responsible for supporting the V coenzyme conformation reported for this enzyme [Arjunan et al. (1996) J. Mol. Biol. 256, 590-600]. Changing the isoleucine to valine and threonine decreased the kcat value and shifted the kcat-pH profile to more alkaline values progressively, indicating that the residue at position 415 not only is important for providing the optimal transition state stabilization but also ensures correct alignment of the ionizable groups participating in catalysis. Substitutions to methionine (the residue used in pyruvate oxidase for this purpose) or leucine (the corresponding residue in transketolase) led to greatly diminished kcat values, showing that for each thiamin diphosphate dependent enzyme an optimal hydrophobic side chain evolved to occupy this key position. Computational studies were carried out on the wild-type enzyme and the I415V, I415G, and I415A variants in both the absence and the presence of pyruvate covalently bound to C2 of the thiazolium ring (the latter is a model for the decarboxylation transition state) to determine whether the size of the side chain is critically required to maintain the V conformation. Briefly, there are sufficient conformational constraints from the binding of the diphosphate side chain and three conserved hydrogen bonds to the 4'-aminopyrimidine ring to enforce the V conformation, even in the absence of a large side chain at position 415. There appears to be increased coenzyme flexibility on substitution of Ile415 to Gly in the absence compared with the presence of bound pyruvate, suggesting that entropy contributes to the rate acceleration. The additional CH3 group in Ile compared to Val also provides increased hydrophobicity at the active center, likely contributing to the rate acceleration. The computational studies suggest that direct proton transfer to the 4'-imino nitrogen from the thiazolium C2H is eminently plausible. PMID- 9748346 TI - Cyclophilin and trigger factor from Bacillus subtilis catalyze in vitro protein folding and are necessary for viability under starvation conditions. AB - Cyclophilin (the product of the ppiB gene) and the trigger factor (the product of the tig gene) are the only cytosolic peptidyl-prolyl cis-trans isomerases that are known in Bacillus subtilis. Both enzymes catalyze the in vitro refolding of ribonuclease T1, a reaction that is limited in rate by a prolyl cis/trans isomerization. The efficiency of cyclophilin as a folding catalyst is only modest with a kcat/KM value of 3.8 x 10(4) M-1 s-1, but the trigger factor shows an almost 40-fold higher specific activity with a kcat/KM value of 1.4 x 10(6) M-1 s 1. This high catalytic activity originates from the tight binding to the protein substrate as reflected in both the low KM value of 0.5 microM and in the strong inhibition of the trigger factor by unfolded proteins. By use of a protein folding assay, the concentrations of cyclophilin and the trigger factor in the cytosol of B. subtilis could be determined as 26 and 35 microM, respectively. Together they account for the entire folding activity that is detectable in crude extracts of wild-type B. subtilis cells. The genes encoding cyclophilin and the trigger factor in the B. subtilis chromosome were disrupted individually and simultaneously. Even in combination, these disruptions had no effect on cell viability in rich medium or under several stress conditions, such as heat, osmotic, or oxidative stress. However, in poor medium and, in particular, in the absence of amino acids, the growth of the double mutant strain was strongly decelerated, indicating that the prolyl isomerases become essential for growth under starvation conditions. It is not yet known whether this function relates to the catalysis of the proline-limited folding of essential proteins. PMID- 9748347 TI - Kinetic parameters for the vesicular acetylcholine transporter: two protons are exchanged for one acetylcholine. AB - The vesicular acetylcholine transporter (VAChT) mediates ACh storage in synaptic vesicles by exchanging cytoplasmic ACh with vesicular protons. This study sought to determine the stoichiometry of exchange by analysis of ligand binding and transport kinetics. The effects of different pH values inside and outside, external ACh concentrations, and electrical potential gradients on ACh transport by vesicles isolated from the electric organ of Torpedo were determined using a pH-jump protocol. The equilibrium binding of a high-affinity analogue of ACh is inhibited by protonation with a pKa of 7.4 +/- 0.3. A two-proton model fits the transport data much better than a one-proton model does, and uptake increases at more positive internal electrical potential, as expected for the two-proton model. Thus, the results support the two-proton model. The transport cycle begins with binding of external ACh to outwardly oriented site 2 (KACho = 20 mM) and protonation of inwardly oriented site 1 (pKa1 = 4.73 +/- 0.05). Loaded VAChT reorients quickly (73 000 min-1) and releases ACh to the inside (KAChi = 44 000 mM) and the proton to the outside. Unloaded, internally oriented site 2 binds a proton (pKa2 = 7.0), after which VAChT reorients (150 +/- 20 min-1) in the rate limiting step and releases the proton to the outside to complete the cycle. Rate constants for the reverse direction also were estimated. Two protons provide a thermodynamic driving force beyond that utilized in vivo, which suggests that vesicular filling is regulated. Other phenomena related to VAChT, namely the time required to fill synaptic vesicles, the fractional orientation of the ACh binding site toward cytoplasm, orientational lifetimes, and the rate of nonquantal release of ACh from cholinergic nerve terminals, were computer-simulated, and the results are compared with physiological observations. PMID- 9748348 TI - Two subunits of heptaprenyl diphosphate synthase of Bacillus subtilis form a catalytically active complex. AB - Heptaprenyl diphosphate synthase of Bacillus subtilis, which participates in the biosynthesis of the side chain of menaquinone-7, is composed of two dissociable subunits, component I and component II, which are encoded by two cistrons in a novel gene cluster of gerC operon [Zhang, Y.-W., et al. (1997) J. Bacteriol. 179, 1417-1419]. This enzyme essentially requires the coexistence of both subunits for its catalysis. Expression vector systems for the two structural genes, gerC1 and gerC3, were constructed separately, and the two components were overproduced in Escherichia coli cells. After purification, their dynamic interactions in forming a catalytically active complex were investigated by gel filtration and immunoblotting analyses. When a mixture of the two components that had been preincubated in the presence of Mg2+ and farnesyl diphosphate was subjected to Superdex 200 gel filtration, a significant elution peak appeared in a region earlier than those observed when they were chromatographed individually. This fraction contained both components I and II, and it corresponded to a molecular mass that is in accord with the sum of the values of the two components. Cross linking studies indicate that the two essential subunits, farnesyl diphosphate, and Mg2+ form a ternary complex which seems to represent a catalytically active state of the heptaprenyl diphosphate synthase. On the other hand, no complex was formed in the presence of isopentenyl diphosphate or inorganic pyrophosphate and Mg2+. A photoaffinity analogue of farnesyl diphosphate was shown to preferentially label the component I protein, suggesting that component I possesses a specific affinity for the allylic substrate. Furthermore, the photoaffinity labeling of component I significantly increased in the presence of component II. The mechanism of catalysis of this unique heteromeric enzyme is understood by assuming that association and dissociation of the two subunits facilitate turnover of catalysis for the synthesis of the amphipathic product from soluble substrates. PMID- 9748350 TI - Synthesis and pharmacology of a new AMPA-kainate receptor agonist with potent convulsant activity. PMID- 9748349 TI - Structure-activity relationship of synthetic phosphoinositolglycans mimicking metabolic insulin action. AB - Phosphoinositolglycan (PIG) molecules have been implicated to stimulate glucose and lipid metabolism in insulin-sensitive cells and tissues in vitro and in vivo. The structural requirements for this partial insulin-mimetic activity remained unclear so far. For establishment of a first structure-activity relationship, a number of PIG compounds were synthesized consisting of the complete or shortened/mutated glycan moiety derived from the structure of the glycosylphosphatidylinositol (GPI) anchor of the GPI-anchored protein, Gce1p, from yeast. The PIG compounds were divided into four classes according to their insulin-mimetic activity in vitro with the typical representatives: compound 41, HO-SO2-O-6Manalpha1(Manalpha1-2)-2Manalpha1 (6-HSO3)- -6Manalpha1-4GluNb eta1 6(D)inositol-1,2-(cyclic)-phosphate; compound 37, HO-PO(H)O-6Manalpha1(Manalpha1 2)-2Manalpha1-6Manal pha1-4GluNbeta1-6( D)inositol-1,2-(cyclic)-phosphate; compound 7, HO-PO(H)O-6Manalpha1-4GluN(1-6(L)inositol-1,2-(cyclic)-ph osp hate; and compound 1, HO-PO(H)O-6Manalpha1-4GluN(1-6(L)inositol. Compounds 41 and 37 stimulated lipogenesis up to 90% (at 20 microM) of the maximal insulin response but with differing concentrations required for 50% activation (EC50 values 2.5 +/ 0.9 vs 4.9 +/- 1.7 microM) as well as glycogen synthase (4.7 +/- 1 vs 9.5 +/- 1.5 microM) and glycerol-3-phosphate acyltransferase (3.5 +/- 0.8 vs 8.0 +/- 1.1 microM). Compound 7 was clearly less potent (20% of the maximal insulin response at 100 microM), whereas compound 1 was almost inactive. This relative ranking in the insulin-mimetic potency between members of the PIG classes (e.g., 41 > 37 >> 7 > 1) was also observed for the (i) activation of glucose transport and glucose transporter isoform 4 translocation in isolated normal and insulin-resistant adipocytes, (ii) inhibition of lipolysis in adipocytes, (iii) stimulation of glucose transport and glycogen synthesis in isolated normal and insulin-resistant diaphragms, and (iv) induction of tyrosine phosphorylation of insulin receptor substrate-1 (IRS-1) in diaphragms. The complete glycan core structure (Man3-GluN) of typical GPI anchors including a mannose side chain and the inositolphosphate moiety was required for maximal insulin-mimetic activity of the PIG compounds with some variations possible with respect to the type of residues coupled to the terminal mannose/inositol as well as the type of linkages involved. These data argue for the potency and specificity of the interaction of PIG molecules with putative signaling component(s) (presumably at the level of the IRS proteins) in adipose and muscle cells which finally lead to insulin-mimetic metabolic activity even in insulin-resistant states. PMID- 9748351 TI - New antipsychotic agents with serotonin and dopamine antagonist properties based on a pyrrolo[2,1-b][1,3]benzothiazepine structure. AB - The development of a synthetic approach to the novel pyrrolo[2, 1 b][1,3]benzothiazepine and its derivatives and their biological evaluation as potential antipsychotic drugs are described. In binding studies these compounds proved to be potent 5-HT2, D2, and D3 receptor ligands. The more potent benzothiazepine (+/-)-3b was resolved into its enantiomers by using HPLC techniques. In vitro testing confirmed that (-)-3b is a more potent D2 receptor ligand, maintaining high affinity for 5-HT2 receptors. In contrast, the (+)-3b enantiomer presents a 35 times higher affinity for 5-HT2 than for dopamine D2 receptors with a similar dopamine D1 receptor affinity to that of (-)-3b. Overall, (+)-3b shows an "atypical" neuroleptic binding profile, while (-)-3b has a more "classical" profile. Furthermore pharmacological and biochemical testing displayed that the novel benzothiazepine (+/-)-3b is able to increase the extracellular levels of dopamine in the rat striatum and causes a dose-related suppression of apomorphine-induced locomotor activity. At low doses (+/-)-3b does not induce catalepsy, showing atypical antipsychotic properties similar to those of olanzapine. These heterocyclic compouds represent new leads for the development of novel antipsychotic drugs with atypical properties. PMID- 9748352 TI - Strategies to target kyotorphin analogues to the brain. AB - The design, synthesis, and pharmacological evaluation of brain-targeted chemical delivery systems (CDS) for a kyotorphin analogue (Tyr-Lys) are described. The brain-targeted compound contains the active peptide in a packaged, disguised form, flanked between the lipophilic cholesteryl ester on the C-terminus and the 1, 4-dihydrotrigonellyl redox targetor, attached to the N-terminus through strategically selected L-amino acid(s) spacer. It was found that for successful brain targeting, the epsilon-amine of Lys needs to be also converted to a lipophilic function. Through sequential enzymatic bioactivation, the Tyr-Lys dipeptide is released in a sustained manner, producing significant and prolonged analgesic activity as demonstrated by the rat tail latency test. An alternate strategy was also employed. Lys was replaced by a redox amino acid pair, Nys+ left and right arrow Nys, the nicotinamide left and right arrow 1,4 dihydronicotinamide analogues of Lys (Nys+ is 2-amino-6-(3-carbamoyl-1 pyridiniumyl)hexanoic acid). The Nys form is lipophilic and facilitates delivery in addition to the C- and N-terminal lipophilic functions. Enzymatic oxidation to Nys+ provides the lock-in, followed by removal of the lipophilic groups, releasing Tyr-Nys+ from the brain-targeted analogue (BTRA). Nys+ was shown to be an effective substitution for Arg or Lys. The activities of the CDS and BTRA, respectively, were antagonized by naloxone, supporting the designed brain targeted processes. The most potent compound is the two-proline spacer containing CDS (CDS-PP), followed by the BTRA. PMID- 9748353 TI - Design and synthesis of new potent C2-symmetric HIV-1 protease inhibitors. Use of L-mannaric acid as a peptidomimetic scaffold. AB - A study on the use of derivatized carbohydrates as C2-symmetric HIV-1 protease inhibitors has been undertaken. L-Mannaric acid (6) was bis-O-benzylated at C-2 and C-5 and subsequently coupled with amino acids and amines to give C2-symmetric products based on C-terminal duplication. Potent HIV protease inhibitors, 28 Ki = 0.4 nM and 43 Ki = 0.2 nM, have been discovered, and two synthetic methodologies have been developed, one whereby these inhibitors can be prepared in just three chemical steps from commercially available materials. A remarkable increase in potency going from IC50 = 5000 nM (23) to IC50 = 15 nM (28) was observed upon exchanging -COOMe for -CONHMe in the inhibitor, resulting in the net addition of one hydrogen bond interaction between each of the two -NH- groups and the HIV protease backbone (Gly 48/148). The X-ray crystal structures of 43 and of 48 have been determined (Figures 5 and 6), revealing the binding mode of these inhibitors which will aid further design. PMID- 9748354 TI - Pyrimidine thioethers: a novel class of HIV-1 reverse transcriptase inhibitors with activity against BHAP-resistant HIV. AB - A series of pyrimidine thioethers was synthesized and evaluated for inhibitory properties against wild-type HIV-1 reverse transcriptase (RT) and an RT carrying the resistance-conferring mutation P236L. Modifications of both the pyrimidine and the functionality attached through the thioether yielded several analogues, which demonstrated activity against both enzyme types, with IC50 values as low as 190 nM against wild-type and 66 nM against P236L RT. Evaluation of a select number of pyrimidine thioethers in cell culture showed that these compounds have excellent activity against HIV-1IIIB-WT and retain good activity against a laboratory-derived HIV-1MF delavirdine-resistant variant. PMID- 9748355 TI - Design of self-coded combinatorial libraries to facilitate direct analysis of ligands by mass spectrometry. AB - The direct analysis of selected components from combinatorial libraries by sensitive methods such as mass spectrometry is potentially more efficient than deconvolution and tagging strategies since additional steps of resynthesis or introduction of molecular tags are avoided. A substituent selection procedure is described that eliminates the mass degeneracy commonly observed in libraries prepared by "split-and-mix" methods, without recourse to high-resolution mass measurements. A set of simple rules guides the choice of substituents such that all components of the library have unique nominal masses. Additional rules extend the scope by ensuring that characteristic isotopic mass patterns distinguish isobaric components. The method is applicable to libraries having from two to four varying substituent groups and can encode from a few hundred to several thousand components. No restrictions are imposed on the manner in which the "self coded" library is synthesized or screened. PMID- 9748356 TI - Inhibition of monoamine oxidase-B by condensed pyridazines and pyrimidines: effects of lipophilicity and structure-activity relationships. AB - A number of condensed pyridazines and pyrimidines were synthesized and tested for their monoamine oxidase-A (MAO-A) and MAO-B inhibitory activity. Their lipophilicity was examined by measuring partition coefficients and RP-HPLC capacity factors, revealing some peculiar electronic and conformational effects. Further insights were obtained by X-ray crystallography and a thermodynamic study of RP-HPLC retention. Structure-activity relations highlighted the main factors determining both selectivity and inhibitory potency. Thus, while most of the condensed pyridazines were reversible inhibitors of MAO-B with little or no MAO-A effects, the pyrimidine derivatives proved to be reversible and selective MAO-A inhibitors. Substituents on the diazine nucleus modulated enzyme inhibition. A QSAR analysis of X-substituted 3-X-phenyl-5H-indeno[1,2-c]pyridazin-5-ones showed lipophilicity to increase MAO-B and not MAO-A inhibitory activity. PMID- 9748357 TI - N'-Phenylindol-3-ylglyoxylohydrazide derivatives: synthesis, structure-activity relationships, molecular modeling studies, and pharmacological action on brain benzodiazepine receptors. AB - A series of N'-phenylindol-3-ylglyoxylohydrazides, isosters of the N-benzylindol 3-ylglyoxylamide derivatives previously described by us, were synthesized and tested for their ability to displace [3H]Ro 15-1788 from bovine brain membranes. These compounds were designed with the aim of obtaining products which could exert an in vivo activity, thanks to a higher hydrosolubility and consequently a better bioavailability. Affinity was restricted to the derivatives unsubstituted in the 5 position of the indole nucleus (1, 6, 9, 12, 15, 18, 23, and 26), with Ki values ranging from 510 to 11 nM. The most active compounds (6, 9, 23, and 29) proved to be effective in antagonizing pentylenetetrazole-induced seizures. Molecular modeling studies were performed to rationalize the lack of affinity of hydrazides with a chloro or a nitro group in the 5 position of the indole nucleus. It was hypothesized that the conformational preference of the hydrazide side chain, characterized by a gauche disposition of lone pairs and substituents about the N-N bond, prevents all hydrazides from binding to the receptor similarly to other classes of indole analogues previously investigated. The potency of 5-H hydrazides was attributed to a binding mode which is not feasible for 5-Cl and 5-NO2 counterparts. This theoretical model of ligand-receptor interaction permitted a more stringent interpretation of structure-affinity relationships of hydrazides and of recently described benzylamide derivatives (Da Settimo et al. J. Med. Chem. 1996, 39, 5083-5091). PMID- 9748358 TI - Melatonin receptor ligands: synthesis of new melatonin derivatives and comprehensive comparative molecular field analysis (CoMFA) study. AB - The CoMFA methodology was applied to melatonin receptor ligands in order to establish quantitative structure-affinity relationships. One hundred thirty-three compounds were considered: they were either collected from literature or newly synthesized in order to gain information about the less explored positions. To this end, various melatonin derivatives were prepared and their affinity for quail optic tecta melatonin receptor was tested. Compounds were aligned on the putative active conformation of melatonin proposed by our previously reported pharmacophore search, and their relative affinities were calculated from the displacement of 2-[125I]-iodomelatonin on different tissues expressing aMT receptors. Compounds were grouped into three sets according to their topology. Subset A: melatonin-like compounds; subset B: N-acyl-2-amino-8-methoxytetralins and related compounds; subset C:N-acyl-phenylalkylamines and related compounds. CoMFA models were derived for each set, using the steric, electrostatic, and lipophilic fields as structural descriptors; the PLS analyses were characterized by good statistical parameters, taking into account the heterogeneity of the binding data, obtained with different experimental protocols. From the CoMFA model for the melatonin-like compounds, besides the well-known positive effect of 2-substitution, a low steric tolerance for substituents in 1, 6, and 7, and a negative effect of electron-rich 4-substituents were observed; the information provided by the newly synthesized compounds was essential for these results. Moreover, a comprehensive model for the 133 compounds, accounting for a common alignment and a common mode of interaction at the melatonin receptor, was derived (Q2 = 0.769, R2 = 0.905). This model validates our previously reported pharmacophore search and offers a clear depiction of the structure-affinity relationships for the melatonin receptor ligands. PMID- 9748359 TI - The frontier orbital phase angles: novel QSAR descriptors for benzene derivatives, applied to phenylalkylamine hallucinogens. AB - A new empirical electronic descriptor, obtained from a molecular orbital calculation and applicable to benzene derivatives, is proposed. It is shown that this descriptor, the frontier orbital phase angle, correlates very strongly with the pharmacological activity in humans of a large series of hallucinogenic phenethylamines. In the largest QSAR study on such hallucinogens yet reported, it is demonstrated that the phase of mixing of degenerate frontier orbitals of benzene to form the frontier orbitals of the drug results in the best electronic descriptor yet found for hallucinogenic activity in phenylalkylamines. PMID- 9748360 TI - Inactivation of S-adenosyl-L-homocysteine hydrolase and antiviral activity with 5',5',6',6'-tetradehydro-6'-deoxy-6'-halohomoadenosine analogues (4' haloacetylene analogues derived from adenosine). AB - Treatment of a protected 9-(5, 6-dideoxy-beta-D-ribo-hex-5-ynofuranosyl)adenine derivative with silver nitrate and N-iodosuccinimide (NIS) and deprotection gave the 6'-iodo acetylenic nucleoside analogue 3c. Halogenation of 3-O-benzoyl-5,6 dideoxy-1, 2-O-isopropylidene-alpha-D-ribo-hex-5-enofuranose gave 6-halo acetylenic sugars that were converted to anomeric 1,2-di-O-acetyl derivatives and coupled with 6-N-benzoyladenine. These intermediates were deprotected to give the 6'-chloro 3a, 6'-bromo 3b, and 6'-iodo 3c acetylenic nucleoside analogues. Iodo compound 3c appears to inactivate S-adenosyl-L-homocysteine hydrolase by a type I ("cofactor depletion") mechanism since complete reduction of enzyme-bound NAD+ to NADH was observed and no release of adenine or iodide ion was detected. In contrast, incubation of the enzyme with the chloro 3a or bromo 3b analogues resulted in release of Cl- or Br- and Ade, as well as partial reduction of E-NAD+ to E-NADH. Compounds 3a, 3b, and 3c were inhibitory to replication of vaccinia virus, vesicular stomatitis virus, parainfluenza-3 virus, and reovirus-1 (3a < 3b < 3c, in order of increasing activity). The antiviral effects appear to correlate with type I mechanism-based inhibition of S-adenosyl-L-homocysteine hydrolase. Mechanistic considerations are discussed. PMID- 9748361 TI - Synthesis and biological activity of certain 4'-thio-D-arabinofuranosylpurine nucleosides. AB - A series of 4'-thio-D-arabinofuranosylpurine nucleosides was prepared and evaluated as potential anticancer agents. The details of a convenient and high yielding synthesis of the carbohydrate precursor 1-O-acetyl-2,3,5-tri-O-benzyl-4 thio-D-arabinofuranose (6) are presented. Proof of structure and configuration at all chiral centers of the nucleosides was obtained through an X-ray crystal structure of 9alpha as well as through NOE experiments on 9beta and 9alpha. All six target compounds were evaluated in a series of human cancer cell lines in culture. Two target compounds, beta anomers with diaminopurine (12) and guanine (16) as the bases, had significant cytotoxicity. One of these compounds (12) was selected for animal studies but was found to have no selectivity at the maximum tolerated dose in the murine colon 36 tumor model. PMID- 9748362 TI - Extended aromatic furan amidino derivatives as anti-Pneumocystis carinii agents. AB - The syntheses of nine new derivatives of 2, 5-bis[4-(N-alkylamidino)phenyl]furans with extended aromatic systems are reported. The interaction of these dicationic furans with poly(dA)poly(dT) and with the duplex oligomers d(CGCGAATTCGCG)2 and d(GCGAATTCGC)2 was determined by Tm measurement, and the effectiveness of these compounds against the immunosuppressed rat model of Pneumocystis carinii was evaluated. At a screening dose of 10 micromol/kg, 4 of the 12 amidino furans described here are more active than the parent compound 1. In general, extension of the aromatic system in the absence of a substitution of the amidino nitrogens resulted in higher affinity for DNA than the parent compound as judged by the larger DeltaTm values and suggests enhanced van der Waals interactions in the amidino furan-DNA complex. Three of the compounds, 3, 5, and 11, yield cysts counts of less than 0.1% of control when administered at a dosage of 10 micromol/kg. Compound 3, which does not have an extended aromatic system, is the most active derivative. Although a direct correlation between anti-P. carinii activity and DNA binding affinity was not observed, all compounds which have significant activity have large DeltaTm values. PMID- 9748363 TI - Selective inhibitors of human lactate dehydrogenases and lactate dehydrogenase from the malarial parasite Plasmodium falciparum. AB - Derivatives of the sesquiterpene 8-deoxyhemigossylic acid (2, 3-dihydroxy-6 methyl-4-(1-methylethyl)-1-naphthoic acid) were synthesized that contained altered alkyl groups in the 4-position and contained alkyl or aralkyl groups in the 7-position. These substituted dihydroxynaphthoic acids are selective inhibitors of human lactate dehydrogenase-H (LDH-H) and LDH-M and of lactate dehydrogenase from the malarial parasite Plasmodium falciparum (pLDH). All inhibitors are competitive with the binding of NADH. Selectivity for LDH-H, LDH M, or pLDH is strongly dependent upon the groups that are in the 4- and 7 positions of the dihydroxynaphthoic acid backbone. Dissociation constants as low as 50 nM were observed, with selectivity as high as 400-fold. PMID- 9748364 TI - Synthesis of reagents for the construction of hypusine and deoxyhypusine peptides and their application as peptidic antigens. AB - Two new synthetic methods which allow access to (2S)-deoxyhypusine, natural (2S,9R)-hypusine, (2S,9S)-hypusine, and deoxyhypusine- and hypusine-containing peptides are described. The methods involve both the construction of a deoxyhypusine reagent in which the alpha-nitrogen protecting group is orthogonal to the N-7 and N-12 protecting groups and an alternate synthesis of our previous hypusine reagent, a synthesis which provides for better stereochemical control at C-9. Synthetic hypusine and deoxyhypusine can be generated from these reagents. The hypusine-containing hexapeptide (Cys-Thr-Gly-Hpu-His-Gly) is conjugated to ovalbumin (OVA), keyhole limpet hemocyanin (KLH), and a bis-maleimide; KLH conjugates are also made with the deoxyhypusine- and lysine-containing hexapeptides. Monoclonal antibodies are generated to the hypusine-containing hexapeptide-OVA conjugate in mice. These are isolated and screened against the hypusine-containing hexapeptide-KLH and hypusine-containing hexapeptide-bis maleimide conjugates, as well as against the deoxyhypusine-containing and lysine containing hexapeptide-KLH conjugates. These antibodies may be useful in localizing intracellular hypusine-containing peptides as well as peptides containing hypusine analogues. PMID- 9748365 TI - Effect of polyamine analogues on hypusine content in JURKAT T-cells. AB - The availability of synthetic hypusine and deoxyhypusine has made it possible to develop analytical methods which allow for the measurement of these compounds in various tissues. The methods involve dansylation of extracts from the pellet remaining after perchloric acid precipitation of cell or tissue homogenates, followed by high-performance liquid chromatography. To demonstrate the utility of this approach, the impact of four polyamine analogues, N1,N11-diethylnorspermine (DENSPM), N1,N14-diethylhomospermine (DEHSPM), 1,6,12-triazadodecane [(4,5) triamine], and 1,7, 13-triazatridecane [(5,5) triamine], on hypusine levels in a human T-cell line (JURKAT) is evaluated. All four analogues are active in controlling cell growth and compete well with spermidine for the polyamine transport apparatus. After 144 h of exposure to JURKAT cells, DENSPM reduces putrescine to below detectable limits and spermidine to 10% of the level in control cells. The other three analogues diminish both putrescine and spermidine to below detectable limits. The effectiveness with which the compounds lower spermine levels is DENSPM > DEHSPM > (4,5) triamine > (5,5) triamine. The analogues decrease the activities of ornithine decarboxylase and S adenosylmethionine decarboxylase in a similar fashion. Of the four polyamines, DENSPM and DEHSPM are potent at lowering intracellular hypusine levels after 144 h: 59 +/- 9% and 73 +/- 12% of control levels, respectively. The other two analogues have marginal effects. PMID- 9748366 TI - Naamidine A is an antagonist of the epidermal growth factor receptor and an in vivo active antitumor agent. AB - The known 2-aminoimidazole alkaloid naamidine A (1) was isolated from a Fijian Leucetta sp. sponge as an inhibitor of the epidermal growth factor (EGF) receptor. The compound exhibited potent ability to inhibit the EGF signaling pathway and is more specific for the EGF-mediated mitogenic response than for the insulin-mediated mitogenic response. Evaluation in an A431 xenograft tumor model in athymic mice indicated that naamidine A exhibited at least 85% growth inhibition at the maximal tolerated dose of 25 mg/kg. Preliminary mechanism of action studies indicate that the alkaloid fails to inhibit the binding of EGF to the receptor and has no effect on the catalytic activity of purified c-src tyrosine kinase. PMID- 9748367 TI - Novel peptidyl phosphorus derivatives as inhibitors of human calpain I. AB - Dipeptidyl phosphorus compounds were synthesized as potential bioisosteric mimics of peptide alpha-ketoesters and alpha-ketoacids. alpha-Ketophosphonate Cbz-Leu Leu-P(O)(OCH3)2 (1b), containing an alpha-ketoester bioisostere, inhibits human calpain I with an IC50 = 0.43 microM. The potency of 1b compares very favorably with that of alpha-ketoester Cbz-Leu-Leu-CO2Et (IC50 = 0.60 microM). Monomethyl ketophosphonate Cbz-Leu-Leu-P(O)(OH)(OCH3) (1a, IC50 = 5.2 microM), an alpha ketoacid mimic, is less potent. Dibutyl and dibenzyl alpha-ketophosphonates 1c,e,f are much less potent calpain inhibitors than dimethyl alpha ketophosphonate 1b. alpha-Ketophosphinate 1g (IC50 = 0.37 microM) and alpha ketophosphine oxide 1h (IC50 = 0.35 microM) are also potent calpain inhibitors. PMID- 9748368 TI - Symplostatin 1: A dolastatin 10 analogue from the marine cyanobacterium Symploca hydnoides. AB - A new solid tumor selective cytotoxic analogue of dolastatin 10 (1) has been isolated from the marine cyanobacterium Symploca hydnoides, collected near Guam. This metabolite has been assigned the trivial name symplostatin 1 (2). This discovery supports the proposal that many compounds isolated from the seahare Dolabella auricularia, the original source of the dolastatins, are of dietary origin. PMID- 9748369 TI - Two new steroidal alkaloids, 20-isoveratramine and verapatuline, from the roots and rhizomes of veratrum patulum AB - Roots and rhizomes of Veratrum patulum L. (Liliaceae), used as a source of the Chinese crude drug "Li-lu", have yielded two new steroidal alkaloids, 20 isoveratramine (1) and verapatuline (2), along with three known alkaloids, veratramine (3), veratrosine (4), and jervine (5). Structures of new alkaloids 1 and 2 were determined to be a C-20 epimer of 3 and N-(methoxycarbonyl)jervine, respectively, by the use of spectral data including 2D NMR. PMID- 9748370 TI - New melampolides from Schkuhria schkuhrioides. AB - The novel melampolides (11R)-11,13-dihydro-schkuhriolide (7), (11S)-11,13-dihydro schkuhriolide (8), and schkuhrioidiol (11), along with the known constituents, frutescin (1), schkuhriolide (2), frutescinic acid (4), allo-schkuhriolide (5), and epoxyschkuhriolide (6) were isolated from the aerial parts of Schkuhria schkuhrioides. The structures of the new compounds were determined by spectroscopic methods. Compounds 1, 2, 4, 5, and 6 displayed no significant cytotoxic or antimicrobial activities. PMID- 9748371 TI - The characterization and structure-activity evaluation of toxic norditerpenoid alkaloids from two Delphinium species. AB - A new N-(methylsuccinimido)anthranoyllycoctonine norditerpenoid alkaloid, geyerline, has been isolated and characterized from extracts of the poisonous larkspur Delphinium glaucum. A previously described norditerpenoid alkaloid, grandiflorine, has also been isolated from Delphinium geyeri. Both alkaloids are closely related structurally to the potent neurotoxin methyllycaconitine, established as the primary toxin in many larkspurs poisonous to cattle. Mouse bioassay tests showed grandiflorine to possess toxicity comparable to methyllycaconitine, while its synthetically derived monoacetate, grandiflorine acetate, and geyerline are significantly less toxic. PMID- 9748372 TI - Anti-AIDS agents. 30. Anti-HIV activity of oleanolic acid, pomolic acid, and structurally related triterpenoids. AB - Oleanolic acid (1) was identified as an anti-HIV principle from several plants, including Rosa woodsii (leaves), Prosopis glandulosa (leaves and twigs), Phoradendron juniperinum (whole plant), Syzygium claviflorum (leaves), Hyptis capitata (whole plant), and Ternstromia gymnanthera (aerial part). It inhibited HIV-1 replication in acutely infected H9 cells with an EC50 value of 1.7 microg/mL, and inhibited H9 cell growth with an IC50 value of 21.8 microg/mL [therapeutic index (T. I.) 12.8]. Pomolic acid, isolated from R. woodsii and H. capitata, was also identified as an anti-HIV agent (EC50 1.4 microg/mL, T. I. 16.6). Although ursolic acid did show anti-HIV activity (EC50 2.0 microg/mL), it was slightly toxic (IC50 6.5 microg/mL, T. I. 3.3). A new triterpene (11) was also isolated from the CHCl3-soluble fraction of R. woodsii, though it showed no anti-HIV activity. The structure of 11 was determined to be 1beta-hydroxy-2 oxopomolic acid by spectral examination. Based on these results, we examined the anti-HIV activity of oleanolic acid- or pomolic acid-related triterpenes isolated from several plants. In addition, we previously demonstrated that derivatives of betulinic acid, isolated from the leaves of S. claviflorum as an anti-HIV principle, exhibited extremely potent anti-HIV activity. Accordingly, we prepared derivatives of oleanolic acid and evaluated their anti-HIV activity. Among the oleanolic acid derivatives, 18 demonstrated most potent anti-HIV activity, with an EC50 value of 0. 0005 microg/mL and a T. I. value of 22 400. PMID- 9748373 TI - Three dolabellanes and a macrolide from the sponge Dysidea sp. from Palau. AB - A specimen of Dysidea sp. from Palau contained three new dolabellane diterpenes 1 3 and an unstable 14-membered macrolide, arenolide (4), which showed modest cytotoxicity. From a chemotaxonomic viewpoint, these metabolites are so unusual that we discuss their possible origin. PMID- 9748375 TI - New glycosides from ajuga decumbens AB - A new phenethyl alcohol glycoside, galactosylmartynoside (1), and a new abietatriene-type diterpene glycoside, ajugaside A (2), were isolated from the whole plants of Ajuga decumbens, together with known phenethyl alcohol glycosides (3 and 4) and iridoid glycosides (5-7). Chemical structures were elucidated on the basis of spectral data. Of these compounds, 8-acetylharpagide (6) exhibited the strongest inhibitory effect on Epstein-Barr virus activation induced by 12-O tetradecanoylphorbol-13-acetate. PMID- 9748374 TI - Further purification and structural analysis of calcium spirulan from Spirulina platensis. AB - An antiviral sulfated polysaccharide, calcium spirulan (Ca-SP), isolated from Spirulina platensis, was subjected to further purification. Ca-SP was found to be composed of rhamnose, 3-O-methylrhamnose (acofriose), 2,3-di-O-methylrhamnose, 3 O-methylxylose, uronic acids, and sulfate. The backbone of Ca-SP consisted of 1,3 linked rhamnose and 1,2-linked 3-O-methylrhamnose units with some sulfate substitution at the 4-position. The polymer was terminated at the nonreducing end by 2,3-di-O-methylrhamnose and 3-O-methylxylose residues. PMID- 9748376 TI - Identification and synthesis of novel chlorinated p-anisylpropanoid metabolites from bjerkandera species AB - Analysis of the EtOAc extracts from the culture medium of Bjerkandera sp. BOS55 and B. fumosa revealed the presence of two novel chlorinated metabolites. Their structures were unambiguously established as erythro-1-(3',5'-dichloro-4' methoxyphenyl)-1, 2-propanediol (2) and 1-(3'-chloro-4'-methoxyphenyl)-3-hydroxy 1-propanone (3) through synthesis of authentic samples and comparison of retention times and mass spectral data. The production of trametol (1) by Bjerkandera sp. BOS55 was also a new finding. PMID- 9748377 TI - New p-terphenyl and polyketide metabolites from the sclerotia of Penicillium raistrickii. AB - Three new p-terphenyls (1-3), a new xanthone derivative (4), and two known fungal metabolites (5 and 6) have been isolated from the sclerotia of Penicillium raistrickii (NRRL 2039). The structures for 3,3"-dihydroxy-6' desmethylterphenyllin (1); 3'-demethoxy-6'-desmethyl-5'-methoxycandidusin B (2); 6'-desmethylcandidusin B (3); and 1,3,5, 6-tetrahydroxy-8-methylxanthone (4) were determined on the basis of HRMS and NMR data. Although compounds 1 and 4 exhibited mild antiinsectan and antibacterial activity, griseofulvin (5) was responsible for most of the activity of the sclerotial extract in dietary assays against the corn earworm Helicoverpa zea. PMID- 9748378 TI - Briarane diterpenes from the Indian Ocean gorgonian Gorgonella umbraculum. AB - Three briarane diterpenes, junceellin (1), praelolide (2), and compound 3 (of which 3 is new), were isolated from the gorgonian Gorgonella umbraculum (EII & Sol), and their structures were established on the basis of their spectral data. PMID- 9748379 TI - An A-type proanthocyanidin from Prunus armeniaca. AB - Roots of Prunus armeniaca yielded a new A-type proanthocyanidin whose structure was assigned as ent-epiafzelechin-3-O-p-hydroxybenzoate-(4alpha-->8, 2alpha-->O- >7)-epiafzelechin (1). The structure of 1 was determined through extensive 1D and 2D NMR studies. PMID- 9748380 TI - Antipruritic dinaphthofuran-7,12-dione derivatives from the pericarp of Impatiens balsamina. AB - Dinaphthofuran-7,12-dione derivatives named balsaminones A (1) and B (2) were isolated from the pericarp of Impatiens balsamina L. together with the known compound 2-methoxy-1,4-naphthoquinone (3). Their structures were elucidated by spectral techniques. These compounds have significant antipruritic activity. PMID- 9748381 TI - Indole alkaloids from the tunicate Aplidium meridianum. AB - Five new indole alkaloids, meridianins A-E (1-5), have been isolated from the tunicate Aplidium meridianum, which was collected at a depth of 100 m near the South Georgia Islands, and their structures were elucidated by spectroscopic techniques. Compounds 2-5 showed cytotoxicity toward murine tumor cell lines. PMID- 9748382 TI - Tolyporphins J and K, two further porphinoid metabolites from the cyanobacterium Tolypothrix nodosa. AB - Two new porphinoids, tolyporphins J (1) and K (2), have been isolated from the terrestrial cyanobacterium, Tolypothrix nodosa (HT-58-2) and identified by NMR and mass spectral analysis. The activities of tolyporphins J and K in cell sensitization and drug accumulation assays for multidrug resistance (MDR) reversal were compared with those of tolyporphin A. Unusual NMR spectroscopic shifts were observed for tolyporphin J (1) in CDCl3. PMID- 9748384 TI - Eletefine, a stephaoxocane alkaloid from cissampelos glaberrima AB - A novel isoquinoline alkaloid bearing an oxocane ring (stephaoxocane type (Kashiwaba, N.; Morooka, S.; Kimura, M.; Ono, M.; Toda, J.; Suzuki, H.; Sano, T. J. Nat. Prod. 1996, 59, 803)), has been isolated from the roots of Cissampelos glaberrima. This compound was given the trivial name eletefine (1) and its structure assigned on the basis of spectroscopic data and conversion to the corresponding ketone (2). PMID- 9748383 TI - Isolation and absolute structures of enantiomeric 1,2-bis(4-hydroxy-3 methoxyphenyl)-1,3-propanediol 1-O-glucosides from the bark of Hovenia trichocarpa. AB - Two 1,2-bis(4-hydroxy-3-methoxyphenyl)-1,3-propanediol 1-O-glucosides, hovetrichosides A (1) and B (2), were isolated from the bark of Hovenia trichocarpa. Their structures were established by extensive NMR experiments and chemical methods. Compounds 1 and 2 were (1R), (2S)-1-(4-hydroxy-3-methoxyphenyl) 2-(4-hydroxy-3-methoxyphenyl)-1, 3-propanediol 1-O-beta-D-glucopyranoside and (1S), (2R)-1-(4-hydroxy-3-methoxyphenyl)-2-(4-hydroxy-3-methoxyphenyl)-1, 3 propanediol 1-O-beta-D-glucopyranoside, respectively. PMID- 9748385 TI - Synthesis of geranyl phenyl ethers based on the cytotoxic monoterpenoids from the liverwort genus trichocolea AB - The synthesis of three geranyl ethers, methyl 4-[(2E)-3, 7-dimethyl-5-oxo-2,6 octadienyloxy]-3-methoxybenzoate (1), methyl 4-[(2E)-3,7-dimethyl-2,6 octadienyloxy]-3-methoxybenzoate (2), and methyl 4-[(2E)-3,7-dimethyl-2,6 octadienyloxy]-3-hydroxybenzoate (3), previously isolated from New Zealand Trichocolea liverworts, is described. Differing reactivity of the hydroxy groups of methyl protocatachuate toward alkylation and the ability of alkyl groups to migrate from oxygen to an ortho-oxygen in these benzene derivatives are noted. PMID- 9748386 TI - Antimalarial principles from Artemisia indica. AB - Activity-guided investigation of Artemisia indica Willd. has led to isolation of exiguaflavone A, exiguaflavone B, maackiain, and 2-(2, 4-dihydroxyphenyl)-5,6 methylenedioxybenzofuran. Exiguaflavones A and B exhibit in vitro antimalarial activities of 4.60 x 10(-6) and 7.05 x 10(-6) g/mL, respectively, against Plasmodium falciparum. PMID- 9748387 TI - A revised structure for crotaramosmin from crotolaria ramosissima AB - The structure of crotaramosmin has been reassigned to 1-(5-hydroxy-2, 2-dimethyl 2H-chromen-6-yl)-3-(4-hydroxyphenyl)-propanone (1) as determined by extensive NMR investigation. PMID- 9748389 TI - UV-Guided isolation of alantrypinone, a novel Penicillium alkaloid. AB - Fumiquinazoline F (1) and alantrypinone (2) have been isolated as the two major metabolites of Penicillium thymicola. The structure of 2, which contains a new ring structure, was elucidated by analysis of spectroscopic data including 2D NMR. The absolute configuration of 2 was established by a single-crystal X-ray diffraction study. PMID- 9748388 TI - Antimicrobial activity of 8-alkyl- and 8-phenyl-substituted berberines and their 12-bromo derivatives. AB - The 8-alkyl- (3-6), 8-phenyl- (7), 12-bromo- (8), 8-alkyl-12-bromo- (9-12), and 12-bromo-8-phenyl- (13) berberine derivatives were prepared and tested for their antimicrobial activity in vitro to evaluate structure-activity relationships. Introduction of the alkyl or phenyl group and the bromine atom into the C-8 and C 12 positions of berberine (1), respectively, led to significant increases of the antimicrobial activity. In both the 8-alkyl- and 8-alkyl-12-bromo-berberines (3-6 and 9-12, respectively), the antibacterial activity increased as the length of the aliphatic chain increased. The exception was the activity against Candida albicans and Escherichia coli, which did not always increase as the alkyl side chain lengthened. Among the compounds tested, 12-bromo-8-n-hexylberberine (12) was 64, 256, 128, 16, and 32 times more active against Staphylococcus aureus, Bacillus subtilis, Salmonella enteritidis, E. coli, and C. albicans, respectively, in comparison to the clinically used berberine. Compound 12 was also found to be 8, 16, and 128 times more active against S. aureus, S. enteritidis, and C.albicans, respectively, than kanamycin sulfate, but was of the same order of activity against B. subtilis, and only one-fourth as active against E. coli. PMID- 9748390 TI - A new isoflavone glycoside from dalbergia nigra AB - The new 5-hydroxy-6, 7-dimethoxy-4'-O-(6-O-D-apio-beta-D-furanosyl-beta-D glucopyranosyl)i soflavone (1) has been isolated from Dalbergia nigra. The structure was elucidated using extensive spectroscopic analysis (1D and 2D NMR, MS, IR, UV). PMID- 9748391 TI - A new iridoid from Scrophularia auriculata ssp. pseudoauriculata. AB - A new iridoid glycoside, scrovalentinoside (1), was isolated from the MeOH extract of the aerial parts of Scrophularia auriculata L. ssp. pseudoauriculata. The structure of the new compound 1 was elucidated as 6-O-(2", 3"-di-O-acetyl-4" O-p-methoxy-cinnamoyl)-alpha-L-rhamnopyranosyl catalpol by spectroscopic methods. The known iridoid glycoside, scropolioside A; two saponins, verbascosaponin A and verbascosaponin; and the phenylethanoid glycoside, verbascoside, were also isolated. PMID- 9748392 TI - New mycalolides from the marine sponge Mycale magellanica and their interconversion. AB - Three new macrolides, 30-hydroxymycalolide A (4), 32-hydroxymycalolide A (5), and 38-hydroxymycalolide B (6), were isolated from the marine sponge Mycale magellanica. Their structures were assigned on the basis of spectroscopic data. They were cytotoxic against L1210 cells with IC50 values of 0.019, 0.013, and 0.015 microg/mL, respectively. Chemical interconversion of the known mycalolides A-C (1-3) with 4-6 established their stereochemical relationships. PMID- 9748393 TI - Isolation, identification, and biological activity of isopersin, a new compound from avocado idioblast oil cells AB - A new compound, (12Z,15Z)-1-hydroxy-4-oxo-heneicosa-12,15-dien-2-yl acetate, isopersin (2), has been isolated from avocado idioblast oil cells. In artificial diet bioassays, 2 showed no effects on either larval survivorship or growth of early-instar beet armyworm Spodoptera exigua. In contrast, the isomeric persin (1), (12Z, 15Z)-1-acetoxy-2-hydroxy-4-oxo-heneicosa-12,15-diene, reduces larval growth at equivalent concentrations (500 &mgr;g g-1). Compound 2 is not very stable and isomerizes readily to 1. Both compounds are acid-labile, rearranging rapidly to alkylfuran 3 in the presence of traces of acid. PMID- 9748394 TI - Novel betaines from the marine sponge Agelas dispar. AB - Three novel betaine alkaloids, called aminozooanemonin (1), pyridinebetaine A (2), and pyridinebetaine B (3), have been isolated from the Caribbean sponge Agelas dispar. Their structures were determined by FABMS, IR, UV, and 1D and 2D NMR spectroscopic experiments. Aminozooanemonin and pyridinebetaine A showed moderate antibacterial activity. PMID- 9748396 TI - Cytotoxic amides from the octocoral telesto riisei PMID- 9748395 TI - Isolation and synthesis of an alpha-malamic acid derivative from Justicia ghiesbreghtiana. AB - A polar extract of leaves of Justicia ghiesbreghtiana yielded N-(2-hydroxy-4,5 dimethoxyphenyl)-(S)-alpha-malamic acid, 1. Incomplete spectral analysis yielded a hypothetical structure, which was then proven by total synthesis. Coupling of the trifluoroacetate of malic anhydride (trifluoroacetoxysuccinic anhydride) with an arylamine provided the key to regiospecific preparation of the alpha- rather than beta-malamic acids. PMID- 9748397 TI - Synthesis of 17,20alpha/beta-dihydroxy-4-pregnen-3-one and 5beta-pregnanes in spermatozoa of primary and 17alpha-methyltestosterone-induced secondary male grouper (Epinephelus coioides). AB - The in vitro metabolism of [3H]17-hydroxyprogesterone by spermatozoa of primary and 17alpha-methyltestosterone (MT)-induced secondary male grouper (Epinephelus coioides) has been examined. With only the labeled precursor, the four predominant 5beta-pregnanes (17, 20alpha-dihydroxy-5beta-pregnan-3-one; 5beta pregnan-3alpha,17, 20alpha-triol; 3alpha,17-dihydroxy-5beta-pregnan-20-one; and 17hydroxy-5beta-pregnan-3,20-dione) accounted for 75% of the total product yield, with the remainder comprising 17, 20alpha-dihydroxy-4-pregnen-3-one (17,20alphaP) and traces (3%) of 17,20beta-dihydroxy-4-pregnen-3-one. This is the first report of the synthesis of these 5beta-pregnanes by teleost spermatozoa. The addition of excess unlabeled precursor caused a marked shift to the synthesis of only 17,20alphaP. The steroidogenic profile in spermatozoa of the primary and secondary males appears similar in both cases. The shift to 17,20alphaP synthesis at high substrate concentration may suggest a gonadotropin (GtH) control of 17, 20alphaP synthesis, involving the competition for 17P between the high-activity, low-capacity 5beta-reductases and the low-activity, high-capacity 20alpha-HSD. Poor spermiation in MT-induced males may thus be due to the lack of 17P arising from low GtH secretion in vivo in the secondary males. PMID- 9748398 TI - Thyroid hormones promote early metamorphosis in grouper (Epinephelus coioides) larvae. AB - The response of grouper larvae to the thyroid hormones, thyroxine (T4) and triiodothyronine (T3), was examined. Two-, 3-, and 4-week-old grouper larvae were reared in seawater containing either T4 or T3 at 0.01, 0.1, and 1 ppm. T4 or T3 induced metamorphosis in all age groups in a dose-dependent manner. Regardless of the size of the larvae, metamorphosis was completed in 2 days in larvae treated with 1 ppm of either T4 or T3; 3-4 days in larvae exposed to 0.1 ppm; and 5-6 days in larvae immersed in 0.01 ppm. None of the fish in the control group completed metamorphosis during this period. Compared with the control fish, survival rates were higher in groups exposed to 0.01 ppm and lower in those exposed to 1 ppm of T3. In 4-week-old larvae, T4 treatment (0.01 to 1.0 ppm) resulted in higher survival compared to the control. These results suggest that a dose of 0.01 ppm is appropriate for acceleration of metamorphosis and improvement of survival in 3- and 4-week-old grouper larvae. A lower dose may be apropriate for earlier stages. PMID- 9748399 TI - Genomic structure and expression sites of three gonadotropin-releasing hormone genes in one species. AB - In the teleost fish, Haplochromis burtoni, gonadotropin-releasing hormone (GnRH) peptide has been localized to three distinct regions in the brain. Each GnRH population is associated with expression of a distinct cDNA as previously described. Here we report the complete genomic sequences encoding these three forms and compare their structural organization, putative regulatory elements, and expression patterns in the body. All three genes share a common structure of four exons: the first exon encodes the 5' untranslated region; the second exon encodes the signal sequence, GnRH decapeptide, and the 5' end of the GnRH associated peptide (GAP); the third exon consists entirely of GAP coding sequence; and the fourth exon encodes the 3' end of GAP and the 3' untranslated region. Each of the three GnRH genes has been shown previously to have a distinct spatial expression pattern in the brain, and here we use reverse transcription and cDNA amplification to demonstrate that each gene is expressed in the body. The gene encoding the releasing form, ?Ser8?GnRH, is expressed in the heart, liver, spleen, kidney, and testis, as well as in the preoptic area. The ?His5Trp7Tyr8?GnRH gene is expressed in the testis as well as in the midbrain. The ?Trp7Leu8?GnRH gene is expressed in the testis and the terminal nerve area. We examined the 500 bp upstream of exon 1 in all three H. burtoni genes and identified putative binding sites for glucocorticoid receptor, androgen receptor, and progesterone receptor, as well as the transcription factors Ap-1 and Sp-1. The genomic sequence encoding the terminal nerve form of GnRH (i.e., ?Trp7Leu8?GnRH) in H. burtoni is remarkably similar to that encoding the presumed releasing form of GnRH in salmonids, especially in the 3' intergenic region. Taken together with phylogenetic and mRNA localization data in salmonids, these data suggest that the gene encoding the releasing form of GnRH in salmonids may not yet be described. PMID- 9748400 TI - Glucagon- and NPY-related peptide-immunoreactive cells in the gut of sea bass (Dicentrarchus labrax L.): a light and electron microscopic study. AB - Glucagon and peptide of the neuropeptide Y (NPY) family immunoreactivities were studied in the gut of sea bass (Dicentrarchus labrax) using antisera against bovine/porcine glucagon, porcine glucagon, glicentin (10-30), bovine pancreatic polypeptide (PP), peptide tyrosine tyrosine (PYY), salmon PYY (sPYY), and NPY. Glucagon-, glicentin-, PYY-, and NPY-immunoreactive (ir) cells were detected in the stomach, and glucagon-, PP-, PYY-, sPYY-, and NPY-ir cells in the intestine. PP, PYY, and NPY immunoreactivities coexisted in intestinal endocrine cells (NPY like peptide containing cells), in some of which there was also glucagon immunoreactivity. Preabsorption tests indicated that different products of the glucagon gene(s) are probably expressed in the stomach and intestine of sea bass and that the peptides belonging to the NPY family in the endocrine cells of the intestine are more similar to NPY than to other peptides of this family. Glucagon ir cells in the stomach, and glucagon/NPY-like containing cells in the intestine, were characterized by conventional and immunogold electron-microscopic techniques. The glucagon cells had secretory granules with a clotted content, the gold particles being observed in both the core and the halo. Glucagon/NPY-like cells showed two types of secretory granules differing in size, both of which were immunogold labeled with anti-NPY and anti-sPYY; the smaller granules were weakly immunogold labeled with anti-glucagon. PMID- 9748401 TI - The expression of insulin-like growth factor-1 during adipogenesis in vivo: effect of thyroxine. AB - The expression of insulin-like growth factor-1 (IGF-1) was examined in subcutaneous (SQ) adipose tissue from 105-day pig fetuses by ribonuclease protection assays and in situ hybridization using a porcine IGF-1 riboprobe. Fetuses were hypophysectomized (hypox) at 70 days of gestation and at 90 days thyroxine (T4) pellets were implanted into some of the hypox fetuses. Fetuses were removed and SQ tissues collected on day 105 of gestation. RNase protection assays followed by scanning laser densitometry revealed that IGF-1 mRNA in SQ adipose tissue in hypox fetuses was significantly decreased to 19.8 +/- 1.2% of control values. T4 treatment increased the expression of IGF-1 by 174 +/- 26.6% of values for hypox fetuses. Using in situ hybridization we showed that fetal hypophysectomy reduced and T4 treatment increased expression of IGF-1 mRNA in the outer SQ adipose tissue layer (P < 0.05). However, T4 treatment after hypox did not influence IGF-1 expression in the inner SQ layer (P > 0.05). IGF-1 was expressed around capillaries, in small fat cells, and in fibroblasts in loose and dense connective tissue. Large fat cells, however, did not express IGF-1. Collectively, we concluded that (1) IGF-1 mRNA was decreased after hypox and increased by T4 treatment in SQ tissue of 105-day fetuses; (2) The expression of the IGF-1 gene was more sensitive to T4 treatment after hypox in outer SQ tissue than in inner SQ tissue; (3) Most stromal cells produced IGF-1 mRNA, and as a result they may influence adipogenesis in the outer layer of SQ adipose tissue; and (4) Once fat cells enlarged, expression of IGF-1 was not detected. Therefore, these studies provide evidence that IGF-1 may mediate the influence of T4 on fetal adipogenesis. PMID- 9748402 TI - Cloning of cDNAs for the pituitary glycoprotein hormone alpha subunit precursor molecules in three amphibian species, Bufo japonicus, Rana catesbeiana, and Cynops pyrrhogaster. AB - We have isolated cDNA clones encoding molecules of putative glycoprotein hormone alpha subunit precursors from their pituitary cDNA libraries for a toad, bullfrog and newt. The insert of the isolated toad cDNA was 562 bp long containing the 5' untranlated, coding and 3'-untranslated regions of 38, 363 and 161 bp, respectively. The insert of the bullfrog cDNA was 604 bp long containing the 5' untranslated, coding and 3'-untranslated regions of 70, 366, and 168 bp, respectively. In the newt, a composite cDNA sequence was estimated from four isolated partial clones. It was 694 bp long and contained the 5'-untranslated region of 89 bp, coding region of 366 bp, and 3'-untranslated region of 91 bp or longer. Amino acid sequences deduced from coding regions of the isolated clones showed that the signal peptides consist of 24 residues and the mature proteins of 96 (toad) or 97 residues (bullfrog and newt). In all three species, an insertion of an amino acid residue was found between residues 26 and 27 of the alpha subunit molecule sequence of all other vertebrate species studied. Interestingly, the percentage identities of the entire amino acid sequence between amphibian and mammalian (or avian) alpha subunits are lower than those between lungfish and mammalian (or avian) alpha subunits. This suggests that amino acid substitutions have occurred more frequently during the course of evolution in the alpha subunit molecule of amphibians than in that of other tetrapod vertebrates, although the biological significance of this is not known. PMID- 9748403 TI - Blocking of KC1O4-induced metamorphosis in premetamorphic sea lampreys by exogenous thyroid hormones (TH); effects of KC1O4 and TH on serum TH concentrations and intestinal thyroxine outer-ring deiodination. AB - Immediately premetamorphic larval sea lampreys (Petromyzon marinus) (>/=120 mm in length) were treated for 4, 8, or 16 weeks with one of two concentrations of either exogenous thyroxine (T4; 1 or 0.5 mg/L) or 3,5,3'-triiodothyronine (T3; 1 or 0.25 mg/L) in the presence or absence of the goitrogen potassium perchlorate (KC1O4; 0.05%) as well as with KC1O4 alone. Larvae from all treatments were examined for signs of metamorphosis, changes in serum T4 and T3 concentrations (serum T4 and serum T3), and changes in intestinal T4 outer-ring (5') deiodination to T3 (T4ORD). KC1O4 depressed both serum T4 and T3 and induced metamorphosis in 80% of larvae treated for 8 weeks or longer. However, neither effect was observed in larvae exposed to KC1O4 combined with either thyroid hormone (TH). These data confirm previous suggestions that exogenous TH blocks KC1O4-induced metamorphosis by elevating serum TH concentrations, and provide evidence that declines in serum TH concentrations are mandatory for precocious metamorphosis. Serum T4, but not serum T3, was elevated following exogenous T4 treatment in the presence or absence of KC1O4. This maintenance of serum T3 at control levels, in the presence of a T4 challenge, was not due to decreases in intestinal T4ORD activity, since T4ORD activity was not affected by any treatments in the study. Exogenous T3 elevated both serum T4 and T3. However, serum T3 in T3-treated larvae decreased with time, suggesting a stringent T3 regulation. Elevation of serum T4 following T3 treatment may have been a result of either inhibition of T4 metabolism, or stimulation of T4 secretion by the endostyle. Based on these results, we conclude that (i) exogenous TH blocks KClO4 induced metamorphosis in sea lampreys and (ii) serum T3 is maintained at control levels despite elevations in serum T4 (its immediate precursor), but this does not involve any changes in intestinal T4ORD activity. PMID- 9748404 TI - Serotonergic stimulation of avian prolactin secretion requires an intact dopaminergic system. AB - In order to ascertain if prolactin (PRL) secretion stimulated by serotonin (5-HT) was mediated by the dopaminergic system, both dopamine (DA) and 5-HT were infused into the third ventricle of anesthetized laying turkeys in conjunction with the D1 DA receptor antagonist R(+)-SCH-23390 HCl or the 5-HT receptor antagonist Mianserin HCl. When infused alone at the rate of 10 nmol/min, both DA and 5-HT increased circulating PRL levels significantly (P < 0. 05) after 20 min (111.0 +/ 19.0 to 334.0 +/- 115.1 ng/ml and 93.3 +/- 13.2 to 173.8 +/- 33.3 ng/ml, respectively). When infused in conjunction with the D1 DA antagonist, the stimulatory effect of both DA and 5-HT on PRL release was significantly suppressed. The 5-HT antagonist completely blocked 5-HT-stimulated PRL release when infused at the rate of 10 nmol/min. When the 5-HT antagonist was co-infused with DA, PRL levels significantly increased (33.5 +/- 3.3 to 112.2 +/- 19.7 ng/ml) after 30 min, an increase that did not differ significantly from that caused by DA alone (63.2 +/- 16.4 to 144.5 +/- 44.1 ng/ml). These data suggest that 5-HT stimulation of PRL secretion requires an intact dopaminergic system and that 5-HT synapses are located further upstream than DA synapses along the PRL releasing pathway within the avian hypothalamus. PMID- 9748405 TI - Involvement of multiple biotransformation processes in the metabolic elimination of testosterone by juvenile and adult fathead minnows (Pimephales promelas). AB - Steroid hormone metabolic clearance pathways are susceptible to induction and suppression resulting from exposure to many xenobiotics. These biochemical effects have the potential to alter steroid hormone homeostasis and, ultimately, steroid hormone-dependent processes such as growth, development, and reproduction. In this study, the metabolic clearance of 17beta-hydroxy-4 androsten-3-one (testosterone) by adult male, adult female, and juvenile fathead minnows (Pimephales promelas) was evaluated. Individual elimination metabolites were identified and rates of metabolite elimination were quantified. Fathead minnows produced a variety of testosterone metabolites including oxido-reduced, hydroxylated, and conjugated derivatives. Metabolites identified by TLC/GC/MS included 4-androstene-3,17-dione (androstenedione), 17beta-hydroxy-5alpha androstan-3-one (5alpha-dihydrotestosterone), 5alpha-androstane-3alpha,17beta diol (3alpha-androstanediol), 5alpha-androstane-3beta,17beta-diol (3beta androstanediol), 17beta-hydroxy-4-androstene-3,11-dione (11-ketotestosterone), 16beta-hydroxy-4-androsten-3-one (16beta-hydroxytestosterone), and 6beta-hydroxy 4-androsten-3-one (6beta-hydroxytestosterone). Testosterone and its metabolites were eliminated in both free and conjugated form. Adult male, adult female, and juvenile fathead minnows eliminated the same profile of testosterone metabolites. However, adult females eliminated androstanediols at a significantly greater rate than did males, and juvenile fish eliminated nearly all testosterone metabolites at greater weight-normalized rates than did adults. These results demonstrate that fathead minnows extensively metabolize testosterone leading to its elimination and provide the foundation upon which the effects of xenobiotics on testosterone metabolism can be assessed. PMID- 9748406 TI - GTH II but not GTH I induces final maturation and the development of maturational competence of oocytes of red seabream in vitro. AB - The effects of red seabream gonadotropins (PmGTH I and PmGTH II) on the induction in vitro of germinal vesicle breakdown (GVBD) and the development of maturational competence (responsiveness to maturation-inducing steroid) were examined in the oocytes of red seabream. PmGTH II was highly effective in inducing GVBD in both maturationally incompetent (45.6 +/- 3.2% GVBD at a concentration of 300 ng/ml) and competent oocytes (42.3 +/- 0.4% GVBD at a concentration of 300 ng/ml). 17,20beta-Dihydroxy-4-pregnen-3-one (DHP, 10 ng/ml) increased the frequency of GVBD induced by PmGTH II. PmGTH I (33, 100, 300, and 900 ng/ml) was unable to induce GVBD at any tested concentration in the presence or absence of DHP. Actinomycin D (1 microgram/ml) and cycloheximide (1 microgram/ml) totally inhibited the PmGTH II-induced GVBD in the presence and in the absence of DHP. Both PmGTH I and PmGTH II stimulated in vitro production of 11-ketotestosterone in sliced testes of red seabream in a similar potency. These results indicate that PmGTH II, but not PmGTH I, induces the final maturation of oocytes, as well as the development of the maturational competence of oocytes, in red seabream. PMID- 9748407 TI - Competitive binding of xenobiotic oestrogens to rat alpha-fetoprotein and to sex steroid binding proteins in human and rainbow trout (Oncorhynchus mykiss) plasma. AB - The ability of a variety of "environmental oestrogens" to compete with radiolabelled steroids to rat alpha-fetoprotein (AFP) and to sex steroid binding proteins was investigated in human and rainbow trout (Oncorhynchus mykiss) plasma. For [3H]oestradiol binding to AFP, diethylstilbestrol and 4 nonylphenoxyacetic acid showed significant competition at concentrations about 100-fold greater than oestradiol (relative binding affinities approximately 1% c.f. oestradiol). All other compounds (phytooestrogens: coumestrol, daidzein, genistein; others: 4-nonylphenol, 4-tert-octylphenol, 4-nonylphenoldiethoxylate, 4-tert-butylphenol, bisphenol-A (Bis-A), bis(2- ethylhexl)phthalate, dioctylphthalate, dibutyl phthalate, 2, 4'DDT (op' enantiomer), 2,4'-DDE (mixed enantiomers), kepone) showed only very weak or no competition (relative binding affinities <<0.1% c.f. oestradiol). The situation for both human and fish plasma was very similar, with only very high concentrations (>>1000 fold more than the natural ligand) of a few of the compounds showing any ability to displace the natural ligand. These results suggest that environmental oestrogenic agents are unlikely to produce biological effects by displacing endogenous steroids from plasma steroid binding proteins unless they are present in very high concentrations. PMID- 9748408 TI - Longitudinal fecal steroid excretion in maned wolves (Chrysocyon brachyurus). AB - This study used a fecal steroid monitoring technique to evaluate reproductive cycles in male (4) and female (15) maned wolves, endangered South American canids. A radiolabeled testosterone infusion on a male revealed a fast and predominantly fecal route of excretion for this steroid. Testosterone was also excreted as eight unidentified metabolites, which was not the primary form of this steroid quantified in our assays. Fecal steroid concentrations (estradiol, E2; progestins, P; testosterone, T) in males and acyclic, nonpregnant (pseudo pregnant), and pregnant females were monitored over four breeding seasons (October-January). Significant differences were detected between longitudinal P profiles of cyclic and acyclic females during estrus, luteal phase, and after birth/end of pseudo-pregnancy. Concentrations of P were also significantly higher in pregnant, compared to nonpregnant females, from proestrus to the end of the pregnant luteal phase. Although levels of T were higher in males than in females throughout the breeding season, no cyclicity in male fecal T concentrations was detected. Values of fecal P, T, and the ratio P/T were useful for differentiating gender and detecting pregnancy in females. Similarities to available data on other canids and the management and conservation implications of these findings were discussed. PMID- 9748409 TI - Characterization of bradykinin-related peptides generated in the plasma of six sarcopterygian species (African lungfish, amphiuma, coachwhip, bullsnake, gila monster, and Gray's monitor). AB - Incubation of heat-denatured plasma from six species occupying different evolutionary positions within the Sarcopterygian lineage [the dipnoan, Protopterus annectens (African lungfish); the urodele, Amphiuma tridactylum (three-toed amphiuma); the colubrid snakes, Pituophis melanoleucus sayi (bullsnake) and Masticophis flagellum (coachwhip); and the lizards Heloderma suspectum (Gila monster) and Varanus Grayi (Gray's monitor)] with trypsin generated bradykinin-related peptides that were detected by radioimmunoassay using an antiserum raised against mammalian bradykinin (BK). The peptides were purified by HPLC and their primary structures were established as lungfish [Tyr1,Gly2,Ala7,Pro8]BK, amphiuma [Phe1,Ile2, Leu5]BK, bullsnake and coachwhip [Val1,Thr6]BK, Gila monster [Leu2, Thr6]BK, and Gray's monitor [Thr6]BK. Monitor BK is identical to the peptide generated in turtle and alligator plasma and coachwhip/bullsnake BK shows one amino acid substitution (Ala1 --> Val) compared with the peptide generated in the plasma of the python. The data provide further evidence for the widespread occurrence of a kallikrein-kininogen system in nonmammalian vertebrates but indicate that the primary structure of BK has been poorly conserved during evolution. PMID- 9748410 TI - Control of vascular smooth muscle cell growth in fowl. AB - In adult domestic fowl, angiotensin (ANG) receptors are present in the vascular smooth muscles (VSM) and in the endothelium, mediating vasorelaxation via endothelium-derived relaxing factor/cGMP. ANG II-induced relaxation is minor in chicks and becomes more marked as they mature but diminishes in adult birds, whereas ANG II neither relaxes nor contracts endothelium-denuded aortae from mature chickens. The present study examines in cultured fowl aortic SM cells whether (1) ANG II stimulates or inhibits VSM cell growth and, if so, whether this growth-stimulatory or -inhibitory effect changes with maturation/aging, and (2) S-nitroso-N-acetylpenicillamine (SNAP), a nitric oxide donor, and cGMP attenuate the basal or stimulated VSM cell growth. [Asp1, Val5]ANG II (native fowl ANG II, 10(-6) M) markedly increased (increase from vehicle control, 226.5%; P < 0.01) [3H]thymidine (Thd) incorporation into DNA of quiescent VSM cells (first subculture) from 6-week-old chicks. This growth-stimulating effect was reduced with age (41.4, 29.6, and 3.2% at 9, 19, and 43 weeks of age, respectively). In contrast, platelet-derived growth factor (PDGF, 20 ng/ml) increased [3H]Thd incorporation similarly in chicks, pullets, and hens. Furthermore, ANG II significantly (45.9%, P < 0.01) attenuated the growth promoting effect of fetal calf serum in cultured VSM cells from 6-week-old chicks. This inhibitory effect also decreased in older birds. ANG II showed neither a growth-stimulatory nor -inhibitory effect in cultured neointimal cells. SNAP attenuated dose dependently (20-60 microM) the basal and PDGF-induced VSM cell growth, whereas cGMP inhibited basal growth only at a high dose (100 microM). These results indicate that in fowl VSM cells, ANG II is mitogenic and antimitogenic in chicks but not in mature birds, suggesting that phenotypic modulation occurs in the ANG receptors/signaling mechanism with maturation/age or in neointimal cells, whereas the mitogenic mechanism via PDGF remains in both young and mature birds. PMID- 9748412 TI - Sixth international symposium on reproductive physiology of fish PMID- 9748411 TI - Characterization of estrous cyclicity in the sable antelope (Hippotragus niger) through fecal progestagen monitoring. AB - Fecal progesterone metabolite monitoring techniques were validated for the sable antelope and used to characterize ovarian cycle dynamics and reproductive seasonality in a captive population at the National Zoological Park's Conservation and Research Center. Hormone was homogeneously distributed within fecal samples. Longitudinal fluctuations in fecal progesterone metabolites were consistent with typical luteal phase patterns and corresponded closely with changes in serum progesterone. The lag time from an im injection of progesterone to peak excretion in feces was 16 h. The pattern of births showed a slight peak in the summer (May-July), but year-round endocrine monitoring of six nonpregnant females showed no evidence of seasonality in ovarian activity. Females exhibited 11-14 estrous cycles per year, averaging 24.2 +/- 0.9 days in length. Luteal and interluteal phases were 18.4 +/- 0.9 and 5.8 +/- 0.4 days in length, respectively. Although only a small number of hippotragine antelope species have been studied, data indicate that they exhibit considerable interspecific variability in estrous cycle length and reproductive seasonality and thus may be a useful group for further investigation of factors regulating fertility. PMID- 9748413 TI - Does oxidized low-density lipoprotein occur in vivo? PMID- 9748414 TI - Cerebrovascular reactivity measurement using near-infrared spectroscopy. PMID- 9748415 TI - Importance of glutamine metabolism in murine macrophages and human monocytes to L arginine biosynthesis and rates of nitrite or urea production. AB - 1. The intermediates of biochemical cycles are commonly utilized for biosynthetic processes; thus at least one intermediate must be replenished de novo to provide constant flux through the cycle. The utilization of L-arginine for NO synthesis in macrophages may thus reduce the concentration of intermediates of the urea cycle. It is possible that a glutamine-utilizing pathway exists in mononuclear phagocytes that may connect with the urea cycle.2. In this paper we report that mouse peritoneal resident and Bacillus Calmette-Guerin (BCG)-activated macrophages and human monocytes are capable of utilizing glutamine at high rates, contain sufficient activity of the enzymes required to convert glutamine to citrulline (and subsequently citrulline to arginine) to account for observed rates of nitrite synthesis in the absence of extracellular L-arginine, and will release nitrite when exposed to intermediates of the proposed glutamine- >arginine pathway.3. The rate of nitrite production (in the absence of extracellular arginine) was reduced by culturing macrophages or monocytes in the presence of the glutaminase inhibitor 6-diazo 5-oxo norleucine.4. The rate and extent of arginase secretion, glutamine utilization, nitrite production (basal and lipopolysaccharide-stimulated) and phosphate-dependent glutaminase activity from BCG-activated macrophages was increased compared with resident cells.5. We suggest that the elevated arginase secretion rates in activated macrophages would effectively increase the intracellular concentration of arginine available for conversion to NO via inducible nitric oxide synthase, the expression of which is known to increase on activation of macrophages or monocytes. Additionally, the rate of L-arginine biosynthesis from glutamine may be increased on immunostimulation of the macrophage. PMID- 9748416 TI - Renal arginine metabolism in fasted rats with subacute short bowel syndrome. AB - 1. Arginine can be produced in the kidney from citrulline. An important source of circulating citrulline is the intestinal breakdown of glutamine. Consequently, partial enterectomy leads to decreased plasma citrulline levels. The aim of the present study was to investigate the effect of diminished arterial citrulline levels on renal arginine production and total-body free arginine pools.2. Renal amino acid metabolism was studied 24 h after 75% small bowel resection in rats fasted overnight (16 h) (n=12; total fast 40 h). Sham-operated (n=9) and non operated 16-h and 40-h fasted controls were studied in parallel (n=8/n=7). During anaesthesia, L-(2, 3-3H)-arginine and para-aminohippuric acid were infused until steady state. Subsequently, arterial and renal venous blood samples were taken. Concentrations of para-aminohippurate and amino acids and specific activity of arginine and citrulline were measured to calculate renal plasma flow, net renal uptake or release, and unidirectional influx or efflux of arginine and citrulline, as well as whole-body arginine turnover.3. Arterial citrulline was decreased in enterectomized rats compared with sham-operated rats (23+/-3 versus 44+/-6 microM). Net renal citrulline uptake and arginine release were almost stoichiometric (-36+/-7 and 38+/-6 nmol.min-1. 100 g-1 body weight respectively in sham-operated rats) and were both diminished by 50% in enterectomized versus sham-operated rats. In all groups, net renal arginine production accounted for less than 10% of whole-body rate of arginine appearance (488 nmol.min-1.100 g-1 body weight in the sham group). Despite decreased net renal citrulline consumption and renal arginine production in enterectomized rats, whole-body rate of arginine appearance and arterial arginine did not change significantly.4. In conclusion, net renal arginine production is reduced 24 h after 75% enterectomy in fasted rats. However, this does not have important effects on whole-body arginine production. PMID- 9748417 TI - Studies in vivo of omega-gliadins in gluten sensitivity (coeliac sprue disease). AB - 1. Highly purified omega-gliadins from wheat were used to challenge gluten sensitized individuals. Characteristic responses by mucosal CD3(+) and gamma delta+ lymphocytes were demonstrated. Each lymphocyte subset showed an increase within 8-12 h post-challenge, indicating a specific response by the rectal mucosa to this gliadin species.2. Available sequence data for the omega-gliadins and homologous proteins from barley and rye indicate a common repeating octapeptide motif (consensus PQQPFPQQ). The results indicate, therefore, that the octapeptide repeat, or a contained sequence such as PQQP, plays an important role in the mucosal immunopathology of gluten sensitivity. PMID- 9748418 TI - Isolation and characterization of the viscous, high-molecular-mass microbial carbohydrate fraction from faeces of healthy subjects and patients with Crohn's disease and the consequences for a therapeutic approach. AB - 1. An earlier study by our group revealed that the viscosity of faeces from patients with Crohn's disease is significantly lower than that of healthy subjects. This is due to low concentrations of a high-molecular-mass carbohydrate, probably of bacterial origin. The cause of this phenomenon might be the impaired barrier function of the gut mucosa. Low viscosity may allow close contact of intestinal contents (bacterial products and toxins) with the intestinal wall. This could play a role in the maintenance of the disease.2. The first aim of this study was to investigate the high-molecular-mass carbohydrate fraction, responsible for viscosity, in detail. We also tried (in a pilot study) to raise the intestinal viscosity of patients with Crohn's disease with the undegradable food additive hydroxypropylcellulose (E463), in an attempt to alleviate clinical symptoms.3. The high-molecular-mass fraction (>300 kDa) responsible for faecal viscosity was sensitive to lysozyme and contained high levels of muramic acid. It was concluded that this material consisted mainly of peptidoglycan polysaccharides and was consequently of bacterial origin. The muramic acid in material from patients with Crohn's disease was 7.5 (1.5-13.9)%, which was less than in healthy subjects [11.4 (8.5-24.1)%; P=0.0004]. Furthermore, viscosity in material from patients with Crohn's disease was found to be half [14.9 (1.0-33.6) cP] of that found in healthy subjects [35.0 (2.7 90.7) cP; P=0.004].4.A daily dose of 1 g of hydroxypropylcellulose caused an increase in faecal viscosity in patients with Crohn's disease (from 1.4 to 2.3 cP) and in healthy subjects (from 4.9 to 7.5 cP). Faecal consistency improved in patients with Crohn's disease (from watery and loose to formed) and the defecation frequency decreased from 3-4 to about 2 times a day. No changes in defecation patterns were found in healthy subjects.5. These data indicate that the high-molecular-mass fraction that is responsible for faecal viscosity is peptidoglycan. Furthermore, a daily dose of a hydroxypropylcellulose solution to increase the viscosity of the intestinal contents of patients with Crohn's disease might be beneficial. This approach merits further study. PMID- 9748419 TI - Inhibition of renal 11beta-hydroxysteroid dehydrogenase in vivo by carbenoxolone in the rat and its relationship to sodium excretion. AB - 1. The type 2 isoform of 11beta-hydroxysteroid dehydrogenase, an enzyme which converts cortisol or corticosterone to inactive 11-ketosteroid metabolites, is thought to be responsible for preventing access of endogenous glucocorticoids to mineralocorticoid receptors in the distal nephron; although direct in vivo evidence for this is still lacking. We have examined whether graded inhibition of renal 11beta-hydroxysteroid dehydrogenase activities in vivo results in corresponding changes in urinary electrolyte excretion due to exposure of mineralocorticoid receptors to circulating endogenous glucocorticoids.2. Anaesthetized rats were infused intravenously with vehicle alone or with one of three doses of carbenoxolone: 0.06, 0.6 or 6 mg/h. After measurement of renal electrolyte excretion, the kidneys were snap-frozen in liquid nitrogen and 11beta hydroxysteroid dehydrogenase activities were measured directly by enzyme assay in the presence of NAD+ or NADP+.3. A dose-dependent inhibition of renal 11beta hydroxysteroid dehydrogenase activities was observed: the low, intermediate and high doses of carbenoxolone causing approximately 50%, 80% and >90% inhibition respectively. Only with the high dose was an effect on renal function observed (decreased fractional Na+ excretion and urinary Na+/K+ ratio).4. The poor correlation between the extent of inhibition of renal 11beta-hydroxysteroid dehydrogenase and altered urinary Na+ excretion, apparent at the lower doses of carbenoxolone, suggests either that 11beta-hydroxysteroid dehydrogenase has considerable functional reserve, or that it may not be the only mechanism determining mineralocorticoid receptor specificity in the distal nephron. PMID- 9748420 TI - A comparison of the development of renal hypertension in male and female rats. AB - 1. The objective of this study was to determine the influence of gender on the development of renal hypertension in Sprague-Dawley rats using the Goldblatt two kidney, one-clip (2K1C) model. In addition, this study examined the effect of ovariectomy upon the development of hypertension in female rats.2. At 10 weeks of age, male, intact female and ovariectomized female rats underwent clipping of the right renal artery or sham operation. Tail-cuff plethysmography was used to monitor the systolic blood pressure of all animals for 7 weeks post-clipping or sham operation. Rats were sub-grouped according to whether or not they developed hypertension (systolic blood pressure >=150 mmHg).3. Within 2 to 3 weeks of clipping, hypertension was induced in only 53% (n=120) of the intact female 2K1C rats, but in 83% (n=18) of the male and 78% (n=18) of the ovariectomized female rats.4. Seven weeks after right renal artery clipping, plasma renin activity was determined in a subset of each group and was found to be 5-6 fold higher in male (17.29+/-4.04 ng angiotensin I.h-1.ml-1) and ovariectomized female (9.71+/-1.25 ng angiotensin I.h-1.ml-1) hypertensive rats compared with their respective normotensive or sham-operated counterparts (3.39+/-0.58 ng angiotensin I.h-1.ml-1 and 1.60+/-0.41 ng angiotensin I.h-1.ml-1 respectively) (P<0.05, analysis of variance). In contrast, the plasma renin activity measured in intact female hypertensive rats was not significantly different from that measured in the corresponding 2K1C normotensive or sham-operated groups.5. These results indicate that the success rate of inducing renal hypertension in Sprague-Dawley rats is higher in males than in intact females. Furthermore, these results suggest that the induction of 2K1C hypertension may be influenced by ovarian hormones. PMID- 9748421 TI - Arrhythmogenic effect of ventriculography in patients with left ventricular dilation and/or hypertrophy. AB - 1. This study examined the effect of an acute injection of contrast medium on generation of arrhythmias in diseased hearts in man.2. Subjects were 100 patients in sinus rhythm undergoing cardiac catheterization in whom good quality echocardiograms could be obtained. The subjects comprised 78 males and 22 females aged 37-83 years.3. Arrhythmia induced by left ventricular angiography ranged from nil to brief bursts of ventricular tachycardia. There was a strongly positive relationship between left ventricular internal dimension in diastole (LVIDd) and induced arrhythmia. Out of 26 patients, 25 developed arrhythmia when LVIDd>=5 cm (96%), but only 24 out of 74 patients developed arrhythmia when LVIDd<5 cm (32%) (P<0.001). In non-dilated hearts where K+<4.0 mmol/l, arrhythmia developed in 100% (10 out of 10) of those with left ventricular hypertrophy (LVH), but in only 40% (8 out of 20) without LVH (P<0. 005). Where K+>=4.0 mmol/l, no arrhythmia occurred in patients with LVH but was present in 52% (31 out of 60) of patients without LVH (P<0.005). There were no relationships with age, end-diastolic pressure, blood pressure, ischaemic heart disease or sex of patient. 4. These data support the view that acute injection of contrast medium in humans induces arrhythmias dependent upon the underlying state of the heart, with potentially complex interplay between left ventricular dimension, hypertrophy and potassium status, supporting similar observations in experimental animals. PMID- 9748422 TI - Administration of albumin to patients with sepsis syndrome: a possible beneficial role in plasma thiol repletion. AB - 1. Albumin is often administered intravenously to critically ill patients as a volume expander, to combat hypoalbuminaemia, and to decrease hyperbilirubinaemia. There is, however, an ongoing debate concerning the therapeutic benefit of the former which is an expensive form of treatment.2. Albumin has several biological functions, in particular as a ligand binder. It also acts as an extracellular transition metal ion-binding and radical-scavenging antioxidant. These functions are influenced by the presence of an exposed thiol group (cys 34) on the surface of the albumin molecule. 3. The ability of infused albumin to influence the plasma thiol pool, and hence antioxidant potential, was investigated in patients with sepsis syndrome.4. Plasma thiol levels rose rapidly after albumin infusion and remained elevated even after plasma albumin levels had declined significantly, due to interstitial leakage. Data are suggestive of some form of thiol exchange in the plasma of these patients between albumin and molecules containing oxidized thiol groups.5. Administration of albumin to patients with sepsis syndrome leads to a sustained increase in plasma thiols. Thiols have several important antioxidant functions, and thiol repletion in these patients, who are known to suffer from oxidative stress, may have beneficial antioxidant effects. Antioxidant repletion may represent an important facet of clinically administered albumin. PMID- 9748423 TI - Porcine hepatic response to sepsis and its amplification by an adrenergic receptor alpha1 agonist and a beta2 antagonist. AB - 1. We investigated the effect of adrenergic receptor stimulation or inhibition on the hepatic ultrastructural changes in a porcine faecal peritonitis model of multi-organ failure. We infused either the alpha1 adrenergic receptor agonist methoxamine or the beta2 adrenergic receptor antagonist ICI 118551 during 8 h of the study.2. Anaesthetized pigs (25-30 kg) were divided into four non-septic groups (control, non-septic, non-septic methoxamine and non-septic ICI 118551) and three septic groups (septic, septic methoxamine and septic ICI 118551).3. Changes in hepatic ultrastructure were measured by morphometric analysis. The septic group was significantly worse than all the non-septic groups. Septic methoxamine and septic ICI 118551 were significantly worse than the septic group.4. Septic methoxamine and septic ICI 118551 had a significantly increased perisinusoidal space; septic methoxamine had significant hepatocyte vacuolation.5. Hepatic ultrastructural changes were independent of hepatic blood flow.6. Septic methoxamine had significant myocardial depression.7. The alpha1 adrenergic receptor agonist methoxamine or the beta2 antagonist ICI 118551 both amplified the hepatic injury normally found during sepsis in our porcine model.8. These findings suggest that during sepsis a protective endogenous beta2 adrenergic receptor-mediated anti-inflammatory response is activated via cell membrane transduction to stimulate the trimeric G-protein complex Gs and activate the second cell messenger cAMP.9. In addition, it is likely that alpha1 adrenergic receptor agonists amplify the inflammatory response by stimulating the cell-surface receptor-linked trimeric G-protein complex to activate Gq and the second cell messenger phospholipase C. PMID- 9748424 TI - Randomized clinical trial comparing the effectiveness of two dietary interventions for patients with hyperlipidaemia. AB - 1. Intervention trials in free-living populations have shown relatively small reductions in risk factors for cardiovascular disease, including lipid levels, and have led some to question whether diet is an effective treatment for hyperlipidaemia. However, behaviour change is a complex process and it is possible that standard intervention methods fail to motivate people sufficiently to comply with dietary advice.2. This study applied motivational interviewing, a style of behaviour change counselling, to dietary education for people with hyperlipidaemia. One-hundred and twenty-one patients with hyperlipidaemia who had been referred to a hospital dietetic department for dietary advice were randomized to receive either standard or motivational dietary interventions for a period of 3 months. Outcomes assessed included dietary knowledge, stage of change, dietary intakes, lipid levels and body mass indices. 3. From baseline, both methods of dietary intervention resulted in self-reported changes in dietary habits and knowledge, statistically significant reductions in intake of total fat (from 32.8% to 28.4%), saturated fat (from 11.4% to 9.2%) and energy intakes [ 239 kcal (-999.98 kJ)/day], and in body mass indices (-0.45 kg/m2). Serum cholesterol did not change significantly in either intervention group.4. Motivational and standard dietary interventions achieved statistically significant changes in reported dietary knowledge and behaviour, and led to a reduction in body weight, but not serum cholesterol. Whether this lack of effect is real or due to subjects overestimating true dietary change cannot be determined. Change in body weight was associated with a reported change in energy intake; this provides some support for there having been a real change in intake. PMID- 9748425 TI - Identification of oxidized low-density lipoprotein in human serum by NMR spectroscopy. AB - 1. In this study we compared the 500 MHz 1H-NMRs from native and oxidized low density lipoproteins. 2. The measurements revealed a characteristic pattern of three resonances in spectra from oxidized, but not from native low-density lipoprotein at 1.17 p.p.m., 1.18 p.p.m. and 1.20 p.p.m. (relative to 3 trimethylsilyl-[2,2,3, 3-2H4]-propionate).3.A quantitative comparison between these resonances in sera from patients with coronary heart disease and healthy control subjects revealed that the intensity was significantly higher in patients with coronary heart disease (1.17 p.p.m.: 0.026+/-0.014 versus 0.015+/-0.019; 1.18 p.p.m.: 0.032+/-0.011 versus 0.017+/-0.021; 1.20 p.p.m.: 0.030+/-0.066 versus 0.010+/-0.005; P<0.05 compared with healthy control subjects for each resonance).4.Fractionation of sera from patients with coronary heart disease revealed that the resonances equal to those obtained from experimentally oxidized low-density lipoprotein are indeed caused by the low-density lipoprotein fraction of the sera.5. When the NMRs from sera were calibrated with oxidized low-density lipoprotein prepared by Cu2+ oxidation, a concentration of 66.5+/-28.6 microgram/ml and 36.3+/-23.7 microgram/ml (P<0.05) was estimated in patients with coronary heart disease and healthy subjects respectively. Elevated levels of oxidized low-density lipoprotein also occurred in those patients with normal serum concentrations of total low-density lipoprotein.6. The study shows a simple method to measure oxidized low-density lipoprotein in human serum and may gain interest to assess the cardiovascular risk factor profiles more completely. PMID- 9748426 TI - Evaluation of potential factors affecting the measurement of cerebrovascular reactivity by near-infrared spectroscopy. AB - 1.Near-infrared (IR) spectroscopy is based on the relative transparency of skin, skull and brain to the light in the near-IR region (700-1100 nm) and on the oxygen-dependent tissue absorption changes of haemoglobin.2. We evaluated the most relevant factors (reproducibility, venous return, age and sex) that might affect reliability of near-IR spectroscopy to test CO2 cerebrovascular reactivity.3.Thirty-four healthy volunteers were enrolled in the study. The protocol consisted of a 3-min baseline, a 3-min hypercapnia (5% CO2 in air) and a 2-min recovery. Transcranial Doppler sonography measurements were simultaneously performed. The CO2 reactivity test was repeated on 27 subjects after 1 h to assess reproducibility. CO2 reactivity was also evaluated at different body positions (supine, 35 degrees Trendelenburg and 35 degrees reverse Trendelenburg), and over a gradual increase of the inspired CO2.4. Changes in near-IR spectroscopy and transcranial Doppler sonography parameters were significantly correlated with variations of end-tidal CO2 (P<0.005). A significant correlation between the reactivity indexes of near-IR spectroscopy parameters and flow velocity was also found (P<0.01). A high reproducibility was also found for deoxyhaemoglobin (rI=0.76), oxyhaemoglobin (rI=0.68) and flow velocity (rI=0.60) reactivity indexes. No significant differences between the reactivity indexes of different body positions were found (P>0.05). The reactivity index of oxyhaemoglobin and deoxyhaemoglobin decreased (P<0.05) and increased (P<0.01) with age respectively.5. We found that near-IR spectroscopy is a reliable and reproducible method for the evaluation of cerebrovascular reactivity and might be considered, after appropriate validation, for the assessment of patients with cerebrovascular disease. PMID- 9748427 TI - Contractile response of isolated human hepatic arteries to alpha-adrenoceptor agonists is not impaired in patients with cirrhosis. AB - 1. Impaired vasoconstriction in animals with cirrhosis is maintained in isolated vessels in vitro, indicating an intrinsic alteration in function or structure of the cells in the vascular wall. This may be due to receptor down-regulation, a defect in post-receptor signal transduction or overproduction of vasodilator compounds. This investigation examined the role of these mechanisms in modulating alpha-adrenoceptor-mediated contraction in hepatic arteries from patients with advanced cirrhosis. 2. Hepatic arteries were obtained from subjects with and without cirrhosis for functional investigation in vitro. Endothelial cell function was assessed using endothelium-dependent (acetylcholine) and independent (3'-morpholinosydnonimine) vasodilators. alpha-Adrenoceptor-mediated contraction was assessed by constructing cumulative concentration-response curves to the alpha1-selective agonist phenylephrine, the non-selective adrenoceptor agonist noradrenaline and the receptor-independent vasoconstrictor potassium chloride. 3. None of the vessels used in this study had an intact endothelium but endothelium independent relaxation was not different in arteries from subject with (79.5+/ 10.16%; n=23) and without (84.45+/-18%; n=20) cirrhosis. Phenylephrine, noradrenaline and potassium chloride produced contractions that were of similar size (P>0.05) in arteries from subjects with (10.10+/-0.97 g, 8.85+/-1.03 g and 8.56+/-0.65 g respectively) and without (10.42+/-1.23 g, 9.58+/-1.39 g and 8. 62+/-0.98 g respectively) cirrhosis. The sensitivities (pD2) of the responses to these agonists were also similar (P<0.05) in arteries from patients with cirrhosis (5.45+/-0.10, 5.60+/-0.12 and 1.57+/-0. 03 respectively) and those from non-cirrhotic donors (5.58+/-0.11, 5. 67+/-0.11 and 1.54+/-0.05 respectively).4. Contraction of the denuded hepatic artery was unaffected by cirrhosis indicating that vascular abnormalities in this condition in man are not due to an intrinsic alteration of smooth muscle cell function in hepatic conduit arteries. PMID- 9748428 TI - Inhaled corticosteroid therapy reduces the early morning peak in cortisol and aldosterone. AB - 1. As mineralocorticoid and adrenocorticoid activity are both under the diurnal control of adrenocorticotropic hormone secretion, we aimed to evaluate whether the normal circadian rhythm of cortisol and aldosterone secretion was suppressed by inhaled corticosteroid therapy.2.Ten normotensive patients with mild-moderate asthma, mean age 24.0 (S.D. 9.8) years and mean arterial pressure 90.7 (9.8) mmHg, were studied in a double-blind, randomized crossover design comparing placebo with fluticasone propionate, 1000 microgram administered twice daily at 08:00 h and 20:00 h. After 5 days of repeated dosing at steady state, measurements were made of plasma cortisol and aldosterone at midnight and 08:00 h.3. With placebo there was a significant (P<0.05) difference between cortisol values at 08:00 h (588.6+/-83.8 nmol/l) and midnight (109.6+/-35.0 nmol/l), whereas after treatment with fluticasone propionate there was no significant difference between levels at 08:00 h (143.3+/-57.4 nmol/l) and midnight (64.3+/ 22.3 nmol/l). For cortisol at 08:00 h there was also a significant (P<0.05) difference between placebo and fluticasone propionate. The same pattern was observed for aldosterone. Plasma aldosterone levels at 08:00 h after treatment with placebo (129.6+/-30.9 nmol/l) were significantly different (P<0. 05) to those seen at midnight (40.4+/-6.2 nmol/l). After treatment with fluticasone propionate, there was no significant difference between levels at midnight (55.4+/-11.7 nmol/l) and 08:00 h (64. 8+/-12.7 nmol/l).4. These results show that inhaled corticosteroid therapy abolishes the circadian rhythm of aldosterone and cortisol secretion. This may have possible implications for patients taking inhaled corticosteroids in terms of the beneficial cardiac effects of suppressing early morning aldosterone. PMID- 9748429 TI - Colicin import into Escherichia coli cells. PMID- 9748430 TI - Genomic analysis reveals chromosomal variation in natural populations of the uncultured psychrophilic archaeon Cenarchaeum symbiosum. AB - Molecular phylogenetic surveys have recently revealed an ecologically widespread crenarchaeal group that inhabits cold and temperate terrestrial and marine environments. To date these organisms have resisted isolation in pure culture, and so their phenotypic and genotypic characteristics remain largely unknown. To characterize these archaea, and to extend methodological approaches for characterizing uncultivated microorganisms, we initiated genomic analyses of the nonthermophilic crenarchaeote Cenarchaeum symbiosum found living in association with a marine sponge, Axinella mexicana. Complex DNA libraries derived from the host-symbiont population yielded several large clones containing the ribosomal operon from C. symbiosum. Unexpectedly, cloning and sequence analysis revealed the presence of two closely related variants that were consistently found in the majority of host individuals analyzed. Homologous regions from the two variants were sequenced and compared in detail. The variants exhibit >99.2% sequence identity in both small- and large-subunit rRNA genes and they contain homologous protein-encoding genes in identical order and orientation over a 28-kbp overlapping region. Our study not only indicates the potential for characterizing uncultivated prokaryotes by genome sequencing but also identifies the primary complication inherent in the approach: the widespread genomic microheterogeneity in naturally occurring prokaryotic populations. PMID- 9748431 TI - A new class of Caulobacter crescentus flagellar genes. AB - Eight Caulobacter crescentus flagellar genes, flmA, flmB, flmC, flmD, flmE, flmF, flmG, and flmH, have been cloned and characterized. These eight genes are clustered in pairs (flmAB, flmCD, flmEF, and flmGH) that appear to be structurally organized as operons. Homology comparisons suggest that the proteins encoded by the flm genes may be involved in posttranslational modification of flagellins or proteins that interact with flagellin monomers prior to their assembly into a flagellar filament. Expression of the flmAB, flmEF, and flmGH operons was shown to occur primarily in predivisional cells. In contrast, the flmCD operon was expressed throughout the cell cycle, with only a twofold increase in predivisional cells. The expression of the three temporally regulated operons was subject to positive regulation by the CtrA response regulator protein. Mutations in class II and III flagellar genes had no significant effect on the expression of the flm genes. Furthermore, the flm genes did not affect the expression of class II or class III flagellar genes. However, mutations in the flm genes did result in reduced synthesis of the class IV flagellin proteins. Taken together, these data indicate that the flm operons belong to a new class of flagellar genes. PMID- 9748432 TI - Isolation of Candida glabrata homologs of the Saccharomyces cerevisiae KRE9 and KNH1 genes and their involvement in cell wall beta-1,6-glucan synthesis. AB - The Candida glabrata KRE9 (CgKRE9) and KNH1 (CgKNH1) genes have been isolated as multicopy suppressors of the tetracycline-sensitive growth of a Saccharomyces cerevisiae mutant with the disrupted KNH1 locus and the KRE9 gene placed under the control of a tetracycline-responsive promoter. Although a cgknh1Delta mutant showed no phenotype beyond slightly increased sensitivity to the K1 killer toxin, disruption of CgKRE9 resulted in several phenotypes similar to those of the S. cerevisiae kre9Delta null mutant: a severe growth defect on glucose medium, resistance to the K1 killer toxin, a 50% reduction of beta-1,6-glucan, and the presence of aggregates of cells with abnormal morphology on glucose medium. Replacement in C. glabrata of the cognate CgKRE9 promoter with the tetracycline responsive promoter in a cgknh1Delta background rendered cell growth tetracycline sensitive on media containing glucose or galactose. cgkre9Delta cells were shown to be sensitive to calcofluor white specifically on glucose medium. In cgkre9 mutants grown on glucose medium, cellular chitin levels were massively increased. PMID- 9748433 TI - New potential cell wall glucanases of Saccharomyces cerevisiae and their involvement in mating. AB - Biotinylation of intact Saccharomyces cerevisiae cells with a nonpermeant reagent (Sulfo-NHS-LC-Biotin) allowed the identification of seven cell wall proteins that were released from intact cells by dithiothreitol (DTT). By N-terminal sequencing, three of these proteins were identified as the known proteins beta exoglucanase 1 (Exg1p), beta-endoglucanase (Bgl2p), and chitinase (Cts1p). One protein was related to the PIR protein family, whereas the remaining three (Scw3p, Scw4p, and Scw10p [for soluble cell wall proteins]) were found to be related to glucanases. Single knockouts of these three potential glucanases did not result in dramatic phenotypes. The double knockout of SCW4 and the homologous gene SCW10 resulted in slower growth, significantly increased release of proteins from intact cells by DTT, and highly decreased mating efficiency when these two genes were disrupted in both mating types. The synergistic behavior of the disruption of SCW4 and SCW10 was partly antagonized by the disruption of BGL2. The data are discussed in terms of a possible counterplay of transglucosidase and glucosidase activities. PMID- 9748434 TI - Isolation and characterization of high-osmolarity-sensitive mutants of fission yeast. AB - For the fission yeast Schizosaccharomyces pombe, adaptation to high-osmolarity medium is mediated by a mitogen-activated protein (MAP) kinase cascade, involving the Wis1 MAP kinase kinase and the Sty1 MAP kinase. The MAP kinase pathway transduces an osmotic signal and accordingly regulates the expression of the downstream target gene (gpd1(+)) that encodes NADH-dependent glycerol-3-phosphate dehydrogenase, in order to adaptively accumulate glycerol inside the cells as an osmoprotectant. We previously characterized a set of high-osmolarity-sensitive S. pombe mutants, including wis1, sty1, and gpd1. In this study, we attempted to further isolate novel osmolarity-sensitive mutants. For some of the mutants isolated, profiles of glycerol production in response to the osmolarity of the growth medium were indistinguishable from that of the wild-type cells, suggesting that they are novel types. They were classified into three distinct types genetically and, thus, were designated hos1, hos2, and hos3 (high osmolarity sensitive) mutants. One of them, the hos1 mutant, was characterized in detail. The hos1 mutant was demonstrated to have a mutational lesion in the known ryh1(+) gene, which encodes a small GTP-binding protein. Disruption of the ryh1(+) gene results not only in osmosensitivity but also in temperature sensitivity for growth. It was also found that the delta ryh1 mutant is severely sterile. These results are discussed with special reference to the osmoadaptation of S. pombe. PMID- 9748435 TI - Sucrose is a nonaccumulated osmoprotectant in Sinorhizobium meliloti. AB - Intracellular accumulation of sucrose in response to lowered water activity seems to occur only in photosynthetic organisms. Here we demonstrate, for the first time, the potent ability of this common sugar, supplied exogenously, to reduce growth inhibition of Sinorhizobium meliloti cells in media of inhibitory osmolarity. Independently of the nature of the growth substrates and the osmotic agent, sucrose appears particularly efficient in promoting the recovery of cytoplasmic volume after plasmolysis. Surprisingly, sucrose is not accumulated by the bacteria at an osmotically efficient level. Instead, it strongly stimulates the accumulation of the main endogenous osmolytes glutamate and N acetylglutaminylglutamine amide (NAGGN). Examining cell volume changes during the hyperosmotic treatment, we found a close correlation between the enhancement of the osmotically active solute pool and the increase in cell volume. Sucrose shares several features with ectoine, another nonaccumulated osmoprotectant for S. meliloti. Overall, osmoregulation in S. meliloti appears to be strongly divergent from that in most bacteria. PMID- 9748436 TI - Limonene-1,2-epoxide hydrolase from Rhodococcus erythropolis DCL14 belongs to a novel class of epoxide hydrolases. AB - An epoxide hydrolase from Rhodococcus erythropolis DCL14 catalyzes the hydrolysis of limonene-1,2-epoxide to limonene-1,2-diol. The enzyme is induced when R. erythropolis is grown on monoterpenes, reflecting its role in the limonene degradation pathway of this microorganism. Limonene-1,2-epoxide hydrolase was purified to homogeneity. It is a monomeric cytoplasmic enzyme of 17 kDa, and its N-terminal amino acid sequence was determined. No cofactor was required for activity of this colorless enzyme. Maximal enzyme activity was measured at pH 7 and 50 degrees C. None of the tested inhibitors or metal ions inhibited limonene 1,2-epoxide hydrolase activity. Limonene-1,2-epoxide hydrolase has a narrow substrate range. Of the compounds tested, only limonene-1,2-epoxide, 1 methylcyclohexene oxide, cyclohexene oxide, and indene oxide were substrates. This report shows that limonene-1,2-epoxide hydrolase belongs to a new class of epoxide hydrolases based on (i) its low molecular mass, (ii) the absence of any significant homology between the partial amino acid sequence of limonene-1,2 epoxide hydrolase and amino acid sequences of known epoxide hydrolases, (iii) its pH profile, and (iv) the inability of 2-bromo-4'-nitroacetophenone, diethylpyrocarbonate, 4-fluorochalcone oxide, and 1, 10-phenanthroline to inhibit limonene-1,2-epoxide hydrolase activity. PMID- 9748438 TI - Poly-beta-hydroxybutyrate turnover in Azorhizobium caulinodans is required for growth and affects nifA expression. AB - Azorhizobium caulinodans is able to fix nitrogen in the free-living state and in symbiosis with the tropical legume Sesbania rostrata. The bacteria accumulate poly-beta-hydroxybutyrate (PHB) under both conditions. The structural gene for PHB synthase, phbC, was inactivated by insertion of an interposon. The mutant strains obtained were devoid of PHB, impaired in their growth properties, totally devoid of nitrogenase activity ex planta (Nif-), and affected in nucleotide pools and induced Fix- nodules devoid of bacteria. The Nif- phenotype was the consequence of the lack of nifA transcription. Nitrogenase activity was partially restored to a phbC mutant by constitutive expression of the nifA gene. However, this constitutive nifA expression had no effect on the nucleotide content or on growth of the phbC mutant. It is suggested that PHB is required for maintaining the reducing power of the cell and therefore the bacterial growth. These observations also suggest a new control of nifA expression to adapt nitrogen fixation to the availability of carbon and reducing equivalents. PMID- 9748437 TI - Mutation analysis of PobR and PcaU, closely related transcriptional activators in acinetobacter. AB - Acinetobacter PobR and PcaU are transcriptional activators that closely resemble each other in primary structure, DNA-binding sites, metabolic modulators, and physiological function. PobR responds to the inducer-metabolite p-hydroxybenzoate and activates transcription of pobA, the structural gene for the enzyme that converts p-hydroxybenzoate to protocatechuate. This compound, differing from p hydroxybenzoate only in that it contains an additional oxygen atom, binds to PcaU and thereby specifically activates transcription of the full set of genes for protocatechuate catabolism. Particular experimental attention has been paid to PobR and PcaU from Acinetobacter strain ADP1, which exhibits exceptional competence for natural transformation. This trait allowed selection of mutant strains in which pobR function had been impaired by nucleotide substitutions introduced by PCR replication errors. Contrary to expectation, the spectrum of amino acids whose substitution led to loss of function in PobR shows no marked similarity to the spectrum of amino acids conserved by the demand for continued function during evolutionary divergence of PobR, PcaU, and related proteins. Surface plasmon resonance was used to determine the ability of mutant PobR proteins to bind to DNA in the pobA-pobR intergenic region. Deleterious mutations that strongly affect DNA binding all cluster in and around the PobR region that contains a helix-turn-helix motif, whereas mutations causing defects in the central portion of the PobR primary sequence do not seem to have a significant effect on operator binding. PCR-generated mutations allowing PobR to mimic PcaU function invariably caused a T57A amino acid substitution, making the helix-turn helix sequence of PobR more like that of PcaU. The mutant PobR depended on p hydroxybenzoate for its activity, but this dependence could be relieved by any of six amino acid substitutions in the center of the PobR primary sequence. Independent mutations allowing PcaU to mimic PobR activity were shown to be G222V amino acid substitutions in the C terminus of the 274-residue protein. Together, the analyses suggest that PobR and PcaU possess a linear domain structure similar to that of LysR transcriptional activators which largely differ in primary structure. PMID- 9748439 TI - Structure and mechanism of action of the protease that degrades small, acid soluble spore proteins during germination of spores of Bacillus species. AB - The germination protease (GPR) of Bacillus megaterium initiates the degradation of small, acid-soluble proteins during spore germination. Trypsin treatment of the 46-kDa GPR zymogen (termed P46) removes an approximately 15-kDa C-terminal domain generating a 30-kDa species (P30) which is stable against further digestion. While P30 is not active, it does autoprocess to a smaller form by cleavage of the same bond cleaved in conversion of P46 to the active 41-kDa form of GPR (P41). Trypsin treatment of P41 cleaves the same bond in the C-terminal part of the protein as is cleaved in the P46-->P30 conversion. While the approximately 29-kDa species generated by trypsin treatment of P41 is active, it is rapidly degraded further by trypsin to small inactive fragments. These results, as well as a thermal melting temperature for P41 which is 13 degreesC lower than that for P46 and the unfolding of P41 at significantly lower concentrations of guanidine hydrochloride than for P46, are further evidence for a difference in tertiary structure between P46 and P41, with P46 presumably having a more compact stable structure. However, circular dichroism spectroscopy revealed no significant difference in the secondary structure content of P46 and P41. The removal of approximately 30% of P46 or P41 without significant loss in enzyme activity localized GPR's catalytic residues to the N-terminal two-thirds of the molecule. This finding, as well as comparison of the amino acid sequences of GPR from three different species, analysis of several site-directed GPR mutants, determination of the metal ion content of purified GPR, and lack of inhibition of P41 by a number of protease inhibitors, suggests that GPR is not a member of a previously described class of protease. PMID- 9748440 TI - Characterization of an A-factor-responsive repressor for amfR essential for onset of aerial mycelium formation in Streptomyces griseus. AB - A-factor (2-isocapryloyl-3R-hydroxymethyl-gamma-butyrolactone) is essential for the initiation of aerial mycelium formation in Streptomyces griseus. amfR is one of the genes which, when cloned on a low-copy-number plasmid, suppresses the aerial mycelium-negative phenotype of an A-factor-deficient mutant of S. griseus. Disruption of the chromosomal amfR gene resulted in complete abolition of aerial mycelium formation, indicating that amfR is essential for the onset of morphogenesis. Cloning and nucleotide sequencing of the region upstream of amfR predicted an operon consisting of orf5, orf4, and amfR. Consistent with this idea, Northern blotting and S1 mapping analyses suggested that these three genes were cotranscribed mainly by a promoter (PORF5) in front of orf5. Furthermore, PORF5 was active only in the presence of A-factor, indicating that it is A-factor dependent. Gel mobility shift assays showed the presence of a protein (AdpB) able to bind PORF5 in the cell extract from an A-factor-deficient mutant but not from the wild-type strain. AdpB was purified to homogeneity and found to bind specifically to the region from -72 to -44 bp with respect to the transcriptional start point. Runoff transcriptional analysis of PORF5 with purified AdpB and an RNA polymerase complex isolated from vegetative mycelium showed that AdpB repressed the transcription in a concentration-dependent manner. It is thus apparent that AmfR as a switch for aerial mycelium formation and AdpB as a repressor for amfR are members in the A-factor regulatory cascade, leading to morphogenesis. PMID- 9748441 TI - Filamentous bacteriophages of Vibrio parahaemolyticus as a possible clue to genetic transmission. AB - We have previously reported the isolation and characterization of two filamentous bacteriophages of Vibrio parahaemolyticus, designated Vf12 and Vf33. In this study, to understand the potential of these phages as tools for genetic transmission, we investigated the gene structures of replicative-form (RF) DNAs of their genomes and the distribution of these DNAs on chromosomal and extrachromosomal DNAs. The 7,965-bp nucleotide sequences of Vf12 and Vf33 were determined. An analysis of the overall gene structures revealed that Vf12 and Vf33 had conserved regions and distinctive regions. The gene organization of their conserved regions was similar to that of CTX phage of Vibrio cholerae and coliphage Ff of Escherichia coli, while their distinctive regions were characteristic of Vf12 and Vf33 phage genomes. Southern blot hybridization testing revealed that the filamentous phage genomes integrated into chromosomal DNA of V. parahaemolyticus at the distinctive region of the phage genome and were also distributed on some plasmids of V. parahaemolyticus and total cellular DNAs of one Vibrio damsela and one nonagglutinable Vibrio strain tested. These results strongly suggest the possibilities of genetic interaction among the bacteriophage Vf12 and Vf33 genomes and chromosomal and plasmid-borne DNAs of V. parahaemolyticus strains and of genetic transmission among strains through these filamentous phages. PMID- 9748442 TI - Expression of the Kdp ATPase is consistent with regulation by turgor pressure. AB - The kdpFABC operon of Escherichia coli encodes the four protein subunits of the Kdp K+ transport system. Kdp is expressed when growth is limited by the availability of K+. Expression of Kdp is dependent on the products of the adjacent kdpDE operon, which encodes a pair of two-component regulators. Studies with kdp-lac fusions led to the suggestion that change in turgor pressure acts as the signal to express Kdp (L. A. Laimins, D. B. Rhoads, and W. Epstein, Proc. Natl. Acad. Sci. USA 78:464-468, 1981). More recently, effects of compatible solutes, among others, have been interpreted as inconsistent with the turgor model (H. Asha and J. Gowrishankar, J. Bacteriol. 175:4528-4537, 1993). We re examined the effects of compatible solutes and of medium pH on expression of Kdp in studies in which growth rate was also measured. In all cases, Kdp expression correlated with the K+ concentration when growth began to slow. Making the reasonable but currently untestable assumptions that the reduction in growth rate by K+ limitation is due to a reduction in turgor and that addition of betaine does not increase turgor, we concluded that all of the data on Kdp expression are consistent with control by turgor pressure. PMID- 9748444 TI - Formation of single-stranded DNA during DNA transformation of Neisseria gonorrhoeae. AB - Neisseria gonorrhoeae is naturally competent for DNA transformation. In contrast to other natural prokaryotic DNA transformation systems, single-stranded donor DNA (ssDNA) has not previously been detected during transformation of N. gonorrhoeae. We have reassessed the physical nature of gonococcal transforming DNA by using a sensitive nondenaturing native blotting technique that detects ssDNA. Consistent with previous analyses, we found that the majority of donor DNA remained in the double-stranded form, and only plasmid DNAs that carried the genus-specific DNA uptake sequence were sequestered in a DNase I-resistant state. However, when the DNA was examined under native conditions, S1 nuclease-sensitive ssDNA was identified in all strains tested except for those bacteria that carried the dud-1 mutation. Surprisingly, ssDNA was also found during transformation of N. gonorrhoeae comA mutants, which suggested that ssDNA was initially formed within the periplasm. PMID- 9748443 TI - Effect of slow growth on metabolism of Escherichia coli, as revealed by global metabolite pool ("metabolome") analysis. AB - Escherichia coli growing on glucose in minimal medium controls its metabolite pools in response to environmental conditions. The extent of pool changes was followed through two-dimensional thin-layer chromatography of all 14C-glucose labelled compounds extracted from bacteria. The patterns of metabolites and spot intensities detected by phosphorimaging were found to reproducibly differ depending on culture conditions. Clear trends were apparent in the pool sizes of several of the 70 most abundant metabolites extracted from bacteria growing in glucose-limited chemostats at different growth rates. The pools of glutamate, aspartate, trehalose, and adenosine as well as UDP-sugars and putrescine changed markedly. The data on pools observed by two-dimensional thin-layer chromatography were confirmed for amino acids by independent analysis. Other unidentified metabolites also displayed different spot intensities under various conditions, with four trend patterns depending on growth rate. As RpoS controls a number of metabolic genes in response to nutrient limitation, an rpoS mutant was also analyzed for metabolite pools. The mutant had altered metabolite profiles, but only some of the changes at slow growth rates were ascribable to the known control of metabolic genes by RpoS. These results indicate that total metabolite pool ("metabolome") analysis offers a means of revealing novel aspects of cellular metabolism and global regulation. PMID- 9748445 TI - CheZ has no effect on flagellar motors activated by CheY13DK106YW. AB - The behaviors of both cheZ-deleted and wild-type cells of Escherichia coli were found to be very sensitive to the level of expression of CheZ, a protein known to accelerate the dephosphorylation of the response regulator CheY-phosphate (CheY P). However, cells induced to run and tumble by the unphosphorylated mutant protein CheY13DK106YW (CheY**) failed to respond to CheZ, even when CheZ was expressed at high levels. Therefore, CheZ neither affects the flagellar motors directly nor sequesters CheY**. In in vitro cross-linking studies, CheY** promoted trimerization of CheZ to the same extent as wild-type CheY but failed to induce the formation of complexes of higher molecular weight observed with CheY P. Also, CheY** could be cross-linked to FliM, the motor receptor protein, nearly as well as CheY-P. Thus, to CheZ, CheY** looks like CheY, but to FliM, it looks like CheY-P. PMID- 9748446 TI - hrcA, encoding the repressor of the groEL genes in Streptomyces albus G, is associated with a second dnaJ gene. AB - Expression of the principal chaperones of the heat shock stimulon of Streptomyces albus G are under the negative control of different repressors. The dnaK operon is regulated by hspR, the last gene of the operon (dnaK-grpE-dnaJ-hspR). hsp18, encoding a member of the small heat shock protein family, is regulated by orfY, which is in the opposite orientation upstream of hsp18. The groES-groEL1 operon and the groEL2 gene are regulated differently. They present tandem copies of the CIRCE element found in the 5' region of many heat shock genes and shown to act in Bacillus subtilis as an operator for a repressor encoded by hrcA (hrc stands for heat regulation at CIRCE). We report the identification in S. albus of a new heat shock operon containing hrcA and dnaJ homologs. Disruption of hrcA increased the transcription of the groES-groEL1 operon and of the groEL2 gene. These features were lost when the mutant was complemented in trans by an intact copy of hrcA. Despite considerable accumulation of the GroE chaperones in the hrcA mutant, there was no effect on formation of the aerial mycelium and sporulation, indicating that neither hrcA nor the level of groE gene expression is directly involved in the regulation of Streptomyces morphological differentiation. PMID- 9748447 TI - Rare homologous gene targeting in Histoplasma capsulatum: disruption of the URA5Hc gene by allelic replacement. AB - URA5 genes encode orotidine-5'-monophosphate pyrophosphorylase (OMPpase), an enzyme involved in pyrimidine biosynthesis. We cloned the Histoplasma capsulatum URA5 gene (URA5Hc) by using a probe generated by PCR with inosine-rich primers based on relatively conserved sequences in OMPpases from other organisms. Transformation with this gene restored uracil prototrophy and OMPpase activity to UV-mutagenized ura5 strains of H. capsulatum. We attempted to target the genomic URA5 locus in this haploid organism to demonstrate homologous allelic replacement with transforming DNA, which has not been previously done in H. capsulatum and has been challenging in some other pathogenic fungi. Several strategies commonly used in Saccharomyces cerevisiae and other eukaryotes were unsuccessful, due to the frequent occurrence of ectopic integration, linear plasmid formation, and spontaneous resistance to 5-fluoroorotic acid, which is a selective agent for URA5 gene inactivation. Recent development of an efficient electrotransformation system and of a second selectable marker (hph, conferring hygromycin B resistance) for this fungus enabled us to achieve allelic replacement by using transformation with an insertionally inactivated Deltaura5Hc::hph plasmid, followed by dual selection with hygromycin B and 5-fluoroorotic acid, or by screening hygromycin B-resistant transformants for uracil auxotrophy. The relative frequency of homologous gene targeting was approximately one allelic replacement event per thousand transformants. This work demonstrates the feasibility but also the potential challenge of gene disruption in this organism. To our knowledge, it represents the first example of experimentally directed allelic replacement in H. capsulatum, or in any dimorphic systemic fungal pathogen of humans. PMID- 9748449 TI - A complex control system for transcriptional activation from the sid promoter of bacteriophage P4. AB - The sid gene promoter (Psid), which controls expression of the late genes from satellite phage P4, is activated by a unique class of small DNA-binding proteins. The activators from both satellite and helper phages stimulate transcription from Psid. These activators bind to sites centered at position -55 in all the helper and satellite phage late promoters. P4 Psid is unique in that it has an additional activator binding site centered at position -18 (site II). We have constructed a mutant of site II that no longer binds activators. Transcription under the control of satellite phage activators is increased by the site II mutation. In contrast, helper phage activators do not show this increase in transcription from Psid mutated at site II. Competition gel shift analysis reveals that the P4 satellite phage activator, Delta, binds eightfold better to site II than to site I. The products of the sid transcription unit are needed only when a helper phage is present; thus, the satellite phage activators repress transcription until the helper is present to supply a nonrepressing activator. PMID- 9748448 TI - Induction of Ca2+-calmodulin signaling by hard-surface contact primes Colletotrichum gloeosporioides conidia to germinate and form appressoria. AB - Hard-surface contact primes the conidia of Colletotrichum gloeosporioides to respond to plant surface waxes and a fruit-ripening hormone, ethylene, to germinate and form the appressoria required for infection of the host. Our efforts to elucidate the molecular events in the early phase of the hard-surface contact found that EGTA (5 mM) and U73122 (16 nM), an inhibitor of phospholipase C, inhibited (50%) germination and appressorium formation. Measurements of calmodulin (CaM) transcripts with a CaM cDNA we cloned from C. gloeosporioides showed that CaM was induced by hard-surface contact maximally at 2 h and then declined; ethephon enhanced this induction. The CaM antagonist, compound 48/80, completely inhibited conidial germination and appressorium formation at a concentration of 3 microM, implying that CaM is involved in this process. A putative CaM kinase (CaMK) cDNA of C. gloeosporioides was cloned with transcripts from hard-surface-treated conidia. A selective inhibitor of CaMK, KN93 (20 microM), inhibited (50%) germination and appressorium formation, blocked melanization, and caused the formation of abnormal appressoria. Scytalone, an intermediate in melanin synthesis, reversed the inhibition of melanization but did not restore appressorium formation. The phosphorylation of 18- and 43-kDa proteins induced by hard-surface contact and ethephon was inhibited by the treatment with KN93. These results strongly suggest that hard-surface contact induces Ca2+-calmodulin signaling that primes the conidia to respond to host signals by germination and differentiation into appressoria. PMID- 9748450 TI - Degradation of 2,4,6-trichlorophenol by Phanerochaete chrysosporium: involvement of reductive dechlorination. AB - Under secondary metabolic conditions, the lignin-degrading basidiomycete Phanerochaete chrysosporium mineralizes 2,4, 6-trichlorophenol. The pathway for the degradation of 2,4, 6-trichlorophenol has been elucidated by the characterization of fungal metabolites and oxidation products generated by purified lignin peroxidase (LiP) and manganese peroxidase (MnP). The multistep pathway is initiated by a LiP- or MnP-catalyzed oxidative dechlorination reaction to produce 2,6-dichloro-1,4-benzoquinone. The quinone is reduced to 2,6-dichloro 1,4-dihydroxybenzene, which is reductively dechlorinated to yield 2-chloro-1,4 dihydroxybenzene. The latter is degraded further by one of two parallel pathways: it either undergoes further reductive dechlorination to yield 1, 4-hydroquinone, which is ortho-hydroxylated to produce 1,2, 4-trihydroxybenzene, or is hydroxylated to yield 5-chloro-1,2, 4-trihydroxybenzene, which is reductively dechlorinated to produce the common key metabolite 1,2,4-trihydroxybenzene. Presumably, the latter is ring cleaved with subsequent degradation to CO2. In this pathway, the chlorine at C-4 is oxidatively dechlorinated, whereas the other chlorines are removed by a reductive process in which chlorine is replaced by hydrogen. Apparently, all three chlorine atoms are removed prior to ring cleavage. To our knowledge, this is the first reported example of aromatic reductive dechlorination by a eukaryote. PMID- 9748451 TI - Roles of the Escherichia coli small heat shock proteins IbpA and IbpB in thermal stress management: comparison with ClpA, ClpB, and HtpG In vivo. AB - We have constructed an Escherichia coli strain lacking the small heat shock proteins IbpA and IbpB and compared its growth and viability at high temperatures to those of isogenic cells containing null mutations in the clpA, clpB, or htpG gene. All mutants exhibited growth defects at 46 degrees C, but not at lower temperatures. However, the clpA, htpG, and ibp null mutations did not reduce cell viability at 50 degrees C. When cultures were allowed to recover from transient exposure to 50 degrees C, all mutations except Deltaibp led to suboptimal growth as the recovery temperature was raised. Deletion of the heat shock genes clpB and htpG resulted in growth defects at 42 degrees C when combined with the dnaK756 or groES30 alleles, while the Deltaibp mutation had a detrimental effect only on the growth of dnaK756 mutants. Neither the overexpression of these heat shock proteins nor that of ClpA could restore the growth of dnaK756 or groES30 cells at high temperatures. Whereas increased levels of host protein aggregation were observed in dnaK756 and groES30 mutants at 46 degreesC compared to wild-type cells, none of the null mutations had a similar effect. These results show that the highly conserved E. coli small heat shock proteins are dispensable and that their deletion results in only modest effects on growth and viability at high temperatures. Our data also suggest that ClpB, HtpG, and IbpA and -B cooperate with the major E. coli chaperone systems in vivo. PMID- 9748452 TI - ClpB in a cyanobacterium: predicted structure, phylogenetic relationships, and regulation by light and temperature. AB - The sequence of a genomic clone encoding a 100-kDa stress protein of Plectonema boryanum (p-ClpB) was determined. The predicted polypeptide contains two putative ATPase regions located within two highly conserved domains (N1 and N2), a spacer region that likely forms a coiled-coil domain, and a highly conserved consensus CK2 phosphorylation domain. The coiled-coil region and the putative site of phosphorylation are not unique to p-ClpB; they are present in all ClpB sequences examined and are absent from the ClpB paralogs ClpA, ClpC, ClpX, and ClpY. Small quantities of a 4.5-kb p-clpB transcript and 110-kDa cytosolic p-ClpB protein were detected in cells grown under optimal conditions; however, increases in the quantities of the transcript and protein were observed in cells grown under excess light and low temperature conditions. Finally, we analyzed ClpA, ClpB, and ClpC sequences from 27 organisms in order to predict phylogenetic relationships among the homologs. We have used this information, along with an identity alignment, to redefine the Clp subfamilies. PMID- 9748453 TI - Succinoglycan is required for initiation and elongation of infection threads during nodulation of alfalfa by Rhizobium meliloti. AB - Rhizobium meliloti Rm1021 must be able to synthesize succinoglycan in order to invade successfully the nodules which it elicits on alfalfa and to establish an effective nitrogen-fixing symbiosis. Using R. meliloti cells that express green fluorescent protein (GFP), we have examined the nature of the symbiotic deficiency of exo mutants that are defective or altered in succinoglycan production. Our observations indicate that an exoY mutant, which does not produce succinoglycan, is symbiotically defective because it cannot initiate the formation of infection threads. An exoZ mutant, which produces succinoglycan without the acetyl modification, forms nitrogen-fixing nodules on plants, but it exhibits a reduced efficiency in the initiation and elongation of infection threads. An exoH mutant, which produces symbiotically nonfunctional high molecular-weight succinoglycan that lacks the succinyl modification, cannot form extended infection threads. Infection threads initiate at a reduced rate and then abort before they reach the base of the root hairs. Overproduction of succinoglycan by the exoS96::Tn5 mutant does not reduce the efficiency of infection thread initiation and elongation, but it does significantly reduce the ability of this mutant to colonize the curled root hairs, which is the first step of the invasion process. The exoR95::Tn5 mutant, which overproduces succinoglycan to an even greater extent than the exoS96::Tn5 mutant, has completely lost its ability to colonize the curled root hairs. These new observations lead us to propose that succinoglycan is required for both the initiation and elongation of infection threads during nodule invasion and that excess production of succinoglycan interferes with the ability of the rhizobia to colonize curled root hairs. PMID- 9748454 TI - Structural organization of virulence-associated plasmids of Yersinia pestis. AB - The complete nucleotide sequence and gene organization of the three virulence plasmids from Yersinia pestis KIM5 were determined. Plasmid pPCP1 (9,610 bp) has a GC content of 45.3% and encodes two previously known virulence factors, an associated protein, and a single copy of IS100. Plasmid pCD1 (70,504 bp) has a GC content of 44.8%. It is known to encode a number of essential virulence determinants, regulatory functions, and a multiprotein secretory system comprising the low-calcium response stimulation that is shared with the other two Yersinia species pathogenic for humans (Y. pseudotuberculosis and Y. enterocolitica). A new pseudogene, which occurs as an intact gene in the Y. enterocolitica and Y. pseudotuberculosis-derived analogues, was found in pCD1. It corresponds to that encoding the lipoprotein YlpA. Several intact and partial insertion sequences and/or transposons were also found in pCD1, as well as six putative structural genes with high homology to proteins of unknown function in other yersiniae. The sequences of the genes involved in the replication of pCD1 are highly homologous to those of the cognate plasmids in Y. pseudotuberculosis and Y. enterocolitica, but their localization within the plasmid differs markedly from those of the latter. Plasmid pMT1 (100,984 bp) has a GC content of 50.2%. It possesses two copies of IS100, which are located 25 kb apart and in opposite orientations. Adjacent to one of these IS100 inserts is a partial copy of IS285. A single copy of an IS200-like element (recently named IS1541) was also located in pMT1. In addition to 5 previously described genes, such as murine toxin, capsule antigen, capsule anchoring protein, etc., 30 homologues to genes of several bacterial species were found in this plasmid, and another 44 open reading frames without homology to any known or hypothetical protein in the databases were predicted. PMID- 9748455 TI - HrpW of Erwinia amylovora, a new harpin that contains a domain homologous to pectate lyases of a distinct class. AB - Harpins, such as HrpN of Erwinia amylovora, are extracellular glycine-rich proteins that elicit the hypersensitive reaction (HR). We identified hrpW of E. amylovora, which encodes a protein similar to known harpins in that it is acidic, rich in glycine and serine, and lacks cysteine. A putative HrpL-dependent promoter was identified upstream of hrpW, and Western blot analysis of hrpL mutants indicated that the production of HrpW is regulated by hrpL. HrpW is secreted via the Hrp (type III) pathway based on analysis of wild-type strains and hrp secretion mutants. When infiltrated into plants, HrpW induced rapid tissue collapse, which required active plant metabolism. The HR-eliciting activity was heat stable and protease sensitive. Thus, we concluded that HrpW is a new harpin. HrpW of E. amylovora consists of two domains connected by a Pro and Ser-rich sequence. A fragment containing the N-terminal domain was sufficient to elicit the HR. Although no pectate lyase activity was detected, the C-terminal region of HrpW is homologous to pectate lyases of a unique class, suggesting that HrpW may be targeted to the plant cell wall. Southern analysis indicated that hrpW is conserved among several Erwinia species, and hrpW, provided in trans, enhanced the HR-inducing ability of a hrpN mutant. However, HrpW did not increase the virulence of a hrpN mutant in host tissue, and hrpW mutants retained the wild type ability to elicit the HR in nonhosts and to cause disease in hosts. PMID- 9748456 TI - The Pseudomonas syringae pv. tomato HrpW protein has domains similar to harpins and pectate lyases and can elicit the plant hypersensitive response and bind to pectate. AB - The host-specific plant pathogen Pseudomonas syringae elicits the hypersensitive response (HR) in nonhost plants and secretes the HrpZ harpin in culture via the Hrp (type III) secretion system. Previous genetic evidence suggested the existence of another harpin gene in the P. syringae genome. hrpW was found in a region adjacent to the hrp cluster in P. syringae pv. tomato DC3000. hrpW encodes a 42. 9-kDa protein with domains resembling harpins and pectate lyases (Pels), respectively. HrpW has key properties of harpins. It is heat stable and glycine rich, lacks cysteine, is secreted by the Hrp system, and is able to elicit the HR when infiltrated into tobacco leaf tissue. The harpin domain (amino acids 1 to 186) has six glycine-rich repeats of a repeated sequence found in HrpZ, and a purified HrpW harpin domain fragment possessed HR elicitor activity. In contrast, the HrpW Pel domain (amino acids 187 to 425) is similar to Pels from Nectria haematococca, Erwinia carotovora, Erwinia chrysanthemi, and Bacillus subtilis, and a purified Pel domain fragment did not elicit the HR. Neither this fragment nor the full-length HrpW showed Pel activity in A230 assays under a variety of reaction conditions, but the Pel fragment bound to calcium pectate, a major constituent of the plant cell wall. The DNA sequence of the P. syringae pv. syringae B728a hrpW was also determined. The Pel domains of the two predicted HrpW proteins were 85% identical, whereas the harpin domains were only 53% identical. Sequences hybridizing at high stringency with the P. syringae pv. tomato hrpW were found in other P. syringae pathovars, Pseudomonas viridiflava, Ralstonia (Pseudomonas) solanacearum, and Xanthomonas campestris. DeltahrpZ::nptII or hrpW::OmegaSpr P. syringae pv. tomato mutants were little reduced in HR elicitation activity in tobacco, whereas this activity was significantly reduced in a hrpZ hrpW double mutant. These features of hrpW and its product suggest that P. syringae produces multiple harpins and that the target of these proteins is in the plant cell wall. PMID- 9748457 TI - Carbon-source-dependent expression of the PalkB promoter from the Pseudomonas oleovorans alkane degradation pathway. AB - Pseudomonas oleovorans GPo1 can metabolize medium-chain-length alkanes by means of an enzymatic system whose induction is regulated by the AlkS protein. In the presence of alkanes, AlkS activates the expression of promoter PalkB, from which most of the genes of the pathway are transcribed. In addition, expression of the first enzyme of the pathway, alkane hydroxylase, is known to be influenced by the carbon source present in the growth medium, indicating the existence of an additional overimposed level of regulation associating expression of the alk genes with the metabolic status of the cell. Reporter strains bearing PalkB-lacZ transcriptional fusions were constructed to analyze the influence of the carbon source on induction of the PalkB promoter by a nonmetabolizable inducer. Expression was most efficient when cells grew at the expense of citrate, decreasing significantly when the carbon source was lactate or succinate. When cells were grown in Luria-Bertani rich medium, PalkB was strongly down-regulated. This effect was partially relieved when multiple copies of the gene coding for the AlkS activator were present and was not observed when the promoter was moved to Escherichia coli, a heterologous genetic background. Possible mechanisms responsible for PalkB regulation are discussed. PMID- 9748458 TI - Ndd, the bacteriophage T4 protein that disrupts the Escherichia coli nucleoid, has a DNA binding activity. AB - Early in a bacteriophage T4 infection, the phage ndd gene causes the rapid destruction of the structure of the Escherichia coli nucleoid. Even at very low levels, the Ndd protein is extremely toxic to cells. In uninfected E. coli, overexpression of the cloned ndd gene induces disruption of the nucleoid that is indistinguishable from that observed after T4 infection. A preliminary characterization of this protein indicates that it has a double-stranded DNA binding activity with a preference for bacterial DNA rather than phage T4 DNA. The targets of Ndd action may be the chromosomal sequences that determine the structure of the nucleoid. PMID- 9748460 TI - Genetic analysis of mecillinam-resistant mutants of Caulobacter crescentus deficient in stalk biosynthesis. AB - Stalk synthesis in Caulobacter crescentus is a developmentally controlled and spatially restricted event that requires the synthesis of peptidoglycan at the stalk-cell body junction. We show that the beta-lactam antibiotic mecillinam prevents stalk synthesis by inhibiting stalk elongation. In addition, mecillinam causes an increase in the diameter of the stalk at the stalk-cell body junction. We describe two mutations that confer resistance to mecillinam and that prevent stalk elongation. These mutations are probably allelic, and they map to a locus previously not associated with stalk synthesis. PMID- 9748459 TI - Cell division inhibition in Salmonella typhimurium histidine-constitutive strains: an ftsI-like defect in the presence of wild-type penicillin-binding protein 3 levels. AB - Histidine-constitutive (Hisc) strains of Salmonella typhimurium undergo cell division inhibition in the presence of high concentrations of a metabolizable carbon source. Filaments formed by Hisc strains show constrictions and contain evenly spaced nucleoids, suggesting a defect in septum formation. Inhibitors of penicillin-binding protein 3 (PBP3) induce a filamentation pattern identical to that of Hisc strains. However, the Hisc septation defect is caused neither by reduced PBP3 synthesis nor by reduced PBP3 activity. Gross modifications of peptidoglycan composition are also ruled out. D-Cycloserine, an inhibitor of the soluble pathway producing peptidoglycan precursors, causes phenotypic suppression of filamentation, suggesting that the septation defect of Hisc strains may be caused by scarcity of PBP3 substrate. PMID- 9748461 TI - Regulation of Escherichia coli secA by cellular protein secretion proficiency requires an intact gene X signal sequence and an active translocon. AB - secA is translationally regulated by the protein secretion proficiency state of the Escherichia coli cell. This regulation was explored by making signal sequence mutations in the gene upstream of secA, gene X, which promotes secA translational coupling. Gene X signal sequence mutants were constitutive for secA expression, while prlA alleles partially restored secA regulation. These results show that interaction of the pre-gene X protein with the translocon is required for proper secA regulation. Furthermore, gene X signal sequence mutations disrupted secA regulation only in the cis configuration. We propose that nascent pre-gene X protein interacts with the translocon during its secretion to constitute the secretion sensor. PMID- 9748462 TI - The gene for 16S rRNA methyltransferase (ksgA) functions as a multicopy suppressor for a cold-sensitive mutant of era, an essential RAS-like GTP-binding protein in Escherichia coli. AB - Era, a Ras-like GTP-binding protein in Escherichia coli, has been shown to be essential for growth. However, its cellular functions still remain elusive. In this study, a genetic screening of an E. coli genomic library was performed to identify those genes which can restore the growth ability of a cold-sensitive mutant, Era(Cs) (E200K), at a restrictive temperature when expressed in a multicopy plasmid. Among eight suppressors isolated, six were located at 1 min of the E. coli genomic map, and the gene responsible for the suppression of Era(Cs) (E200K) was identified as the ksgA gene for 16S rRNA transmethylase, whose mutation causes a phenotype of resistance to kasugamycin, a translation initiation inhibitor. This is the first demonstration of suppression of impaired function of Era by overproduction of a functional enzyme. A possible mechanism of the suppression of the Era cold-sensitive phenotype by KsgA overproduction is discussed. PMID- 9748463 TI - Suppression of the ptsH mutation in Escherichia coli and Salmonella typhimurium by a DNA fragment from Lactobacillus casei. AB - A DNA fragment from Lactobacillus casei that restores growth to Escherichia coli and Salmonella typhimurium ptsH mutants on glucose and other substrates of the phosphoenolpyruvate:carbohydrate phosphotransferase system (PTS) has been isolated. These mutants lack the HPr protein, a general component of the PTS. Sequencing of the cloned fragment revealed the absence of ptsH homologues. Instead, the complementation ability was located in a 120-bp fragment that contained a sequence homologue to the binding site of the Cra regulator from enteric bacteria. Experiments indicated that the reversion of the ptsH phenotype was due to a titration of the Cra protein, which allowed the constitutive expression of the fructose operon. PMID- 9748464 TI - Bradyrhizobium japonicum FixK2, a crucial distributor in the FixLJ-dependent regulatory cascade for control of genes inducible by low oxygen levels. AB - Bradyrhizobium japonicum possesses a second fixK-like gene, fixK2, in addition to the previously identified fixK1 gene. The expression of both genes depends in a hierarchical fashion on the low-oxygen-responsive two-component regulatory system FixLJ, whereby FixJ first activates fixK2, whose product then activates fixK1. While the target genes for control by FixK1 are unknown, there is evidence for activation of the fixNOQP, fixGHIS, and rpoN1 genes and some heme biosynthesis and nitrate respiration genes by FixK2. FixK2 also regulates its own structural gene, directly or indirectly, in a negative way. PMID- 9748465 TI - Identification of multiple sigma54-dependent transcriptional activators in Vibrio cholerae. AB - In the pathogenic bacterium Vibrio cholerae, the alternate sigma factor sigma54 is required for expression of multiple sets of genes, including an unidentified gene(s) necessary for enhanced colonization within the host. To identify sigma54 dependent transcriptional activators involved in colonization, PCR was performed with V. cholerae chromosomal DNA and degenerate primers, revealing six novel and distinct coding sequences with homology to sigma54-dependent activators. One sequence had high homology to the luxO gene of V. harveyi, which in that organism is involved in quorum sensing. Phenotypes of V. cholerae strains containing mutations in each of the six putative sigma54-dependent activator genes identified one as a probable ntrC homologue. None of the mutant strains exhibited a defect in the ability to colonize infant mice, suggesting the presence of additional sigma54-dependent activators not identified by this technique. PMID- 9748466 TI - H-NS regulates DNA repair in Shigella. AB - We report a new role for H-NS in Shigella spp.: suppression of repair of DNA damage after UV irradiation. H-NS-mediated suppression of virulence gene expression is thermoregulated in Shigella, being functional at 30 degrees C and nonfunctional at 37 to 40 degrees C. We find that H-NS-mediated suppression of DNA repair after UV irradiation is also thermoregulated. Thus, Shigella flexneri M90T, incubated at 37 or 40 degrees C postirradiation, shows up to 30-fold higher survival than when incubated at 30 degrees C postirradiation. The hns mutants BS189 and BS208, both of which lack functional H-NS, show a high rate of survival (no repression) whether incubated at 30 or 40 degrees C postirradiation. Suppression of DNA repair by H-NS is not mediated through genes on the invasion plasmid of S. flexneri M90T, since BS176, cured of plasmid, behaves identically to the parental M90T. Thus, in Shigella the nonfunctionality of H-NS permits enhanced DNA repair at temperatures encountered in the human host. However, pathogenic Escherichia coli strains (enteroinvasive and enterohemorrhagic E. coli) show low survival whether incubated at 30 or 40 degrees C postirradiation. E. coli K-12 shows markedly different behavior; high survival postirradiation at both 30 and 40 degrees C. These K-12 strains were originally selected from E. coli organisms subjected to both UV and X irradiation. Therefore, our data suggest that repair processes, extensively described for laboratory strains of E. coli, require experimental verification in pathogenic strains which were not adapted to irradiation. PMID- 9748467 TI - Nucleotide sequence and characterization of cdrA, a cell division-related gene of Helicobacter pylori. AB - We identified cell division-related gene cdrA in Helicobacter pylori HPK5. The putative gene product, CdrA, is a 367-amino-acid polypeptide that exhibited a high level of homology to conserved hypothetical ATP-binding protein HP0066 of H. pylori 26695, except in the N-terminal region, and showed some similarity to the FtsK/SpoIIIE family proteins. We isolated a cdrA-disrupted mutant by allelic exchange mutagenesis. Because of the low transformation frequency, the possibility that a suppressing mutation would be found in the obtained cdrA mutant was discussed. A repressive role for CdrA on cell division was suggested by the observations that the wild-type strain formed filamentous cells in a high salt level medium at early stationary phase, while a cdrA-disrupted mutant did not show such an abnormality. In addition, the wild-type strain adopted coccoid forms in the stationary phase, whereas the cdrA-disrupted mutant remained mostly as short rods. Furthermore, the cdrA-disrupted mutant regained the filamentation phenotype when the intact cdrA gene was introduced by allelic exchange. Taken together, these observations show that the cdrA gene plays an important role in the cell growth of H. pylori. PMID- 9748468 TI - Growth medium-dependent regulation of Myxococcus xanthus fatty acid content is controlled by the esg locus. AB - We compared the cellular fatty acid profiles of Myxococcus xanthus cells grown in either a Casitone-based complex medium or a chemically defined medium. The cells grown in the complex medium had a much higher content of the abundant branched chain fatty acid iso-15:0 and several other branched-chain species. The higher branched-chain fatty acid content of the cells grown in the complex medium was dependent on the esg locus, which encodes the E1alpha and E1beta components of a branched-chain keto acid dehydrogenase (BCKAD) multienzyme complex involved in branched-chain fatty acid biosynthesis. Cells grown in the complex medium were also found to have a higher level of esg transcription and more BCKAD enzyme activity than cells from the chemically defined medium. The level of esg transcription appears to be an important factor in the growth medium-dependent regulation of the M. xanthus branched-chain fatty acid content. PMID- 9748470 TI - Two amino acid residues of transposase contributing to differential transposability of IS1 elements in Escherichia coli. AB - Escherichia coli W3110 contains four types of IS1 elements in the chromosome. Using an insertion element entrapping system, we collected 116 IS1 plasmid insertion mutants, which resulted from a minimum of 26 independent IS1 insertion events. All of them had insertions of IS1 of the IS1A (IS1E and IS1G) type. Inspection of the transposase sequences of the four IS1 types and the IS1 of the resistance plasmid R100 showed that two amino acid residues, His-193 and Leu-217 of transposase, might contribute to differential transposability of IS1 elements in W3110. The two amino acid residues of the transposase in IS1A (IS1E and IS1G) were altered separately by site-directed mutagenesis, and each mutant was found to mediate transposition at a frequency about 30-fold lower than that of IS1A (IS1E and IS1G). Thus, the assumption that His-193 and Leu-217 of transposase contribute to differential transposability of IS1 elements in W3110 was confirmed. PMID- 9748469 TI - Molecular characterization of the complete 23F capsular polysaccharide locus of Streptococcus pneumoniae. AB - The complete DNA sequence of the capsular locus 23F of Streptococcus pneumoniae is presented. The 18.6-kb cps23f locus is composed of 18 open reading frames flanked at the 5' and 3' ends by the genes dexB and aliA, an arrangement similar to those of some of the other identified cps loci. PMID- 9748471 TI - Structure-function relationships in the ezrin family and the effect of tumor associated point mutations in neurofibromatosis 2 protein. AB - Ezrin, radixin and moesin (ERM proteins) link cell adhesion molecules to the cytoskeleton, modulate cell morphology and cell growth and are involved in Rho mediated signal transduction. Merlin, the tumor suppressor in neurofibromatosis 2, is a diverged member of the ezrin family, but its function is at least partially similar to the ERM proteins. In the N-domain, the ezrin family belongs to the band 4.1 superfamily. Secondary structure predictions made separately for the ezrin and band 4.1-tyrosine phosphatase families give a similar pattern for the homologous N-domains, indicating that both families have a similar binding site for the integral membrane proteins. The alpha-domain shows a strong coiled coil prediction, that can be involved in the protein dimerization. The C-terminal actin-binding site in the ERM proteins and the actin-binding helix in the villin headpiece have a common amino acid motif. In merlin, the published tumor associated single amino acid mutations in the N-domain are located in the conserved sites, and they affect mainly the predicted helices and strands, indicating that these mutations cause the disease primarily by disturbing the protein structure. In the alpha- and C-domains, some of the mutations break the helical structures. Some known mutations are observed at a site potentially interacting with cell adhesion molecules. We will also discuss the implications of the evolutionary information and the actin-binding models in the ezrin family. PMID- 9748472 TI - Psychosine blocks quisqualate-induced glutamate excitotoxicity in hippocampal CA sector neurons. AB - Quisqualate is a potent specific agonist for Group 1 metabotropic glutamate receptors (mGluR's), that activate G protein-coupled phospholipase C (PLC) in a molecular signal-transduction mechanism that raises cytoplasmic Ca2+ and, when excessive, damages hippocampal neurons. Psychosine (beta-galactosylsphingosine), a cationic lysosphingolipid occurring naturally in nervous tissues, dose dependently inhibited PLC activation induced by metabotropic alpha 1-adrenergic receptor signaling in cultured rat brain astrocytes in vitro. In the present study, we have tested neuroprotective efficacy of psychosine in vivo, in a rat model of glutamate excitotoxicity induced by intracerebroventricular (i.c.v.) administration of quisqualate. A sublethal i.c.v. dose of quisqualate caused episodes of prolonged akinesia and convulsions, and major damage to pyramidal neurons of the hippocampal CA1 and CA3 sector, but not to granule cell neurons of the dentate gyrus. Prior infusion of psychosine greatly attenuated quisqualate induced behaviors, and fully prevented destruction by quisqualate of vulnerable hippocampal neurons. Psychosine may prove useful in prophylaxis of neurodegenerative disorders that arise from intensive hippocampal Group 1 mGluR stimulation. PMID- 9748474 TI - The C-terminus of human bleomycin hydrolase is required for protection against bleomycin-induced chromosomal damage. AB - Mammalian bleomycin hydrolases (BH) are enzymes with proven exopeptidase activity responsible for deamidation of the beta-aminoalanine moiety in bleomycin and are thought to limit the therapeutic efficacy of the drug. We have recently determined that the highly conserved BH-like domain in the C-terminus of human bleomycin hydrolase (hBH) is critical both for in vitro aminopeptidase and bleomycin metabolizing activities. To determine if hBH protects mammalian cells against bleomycin clastogenic effect, we transfected CHO cells with plasmids encoding hBH or C-terminal truncated forms and evaluated the level of chromatid breaks after bleomycin exposure. CHO cells expressing hBH had 50% less chromatid breaks after bleomycin treatment compared with mock transfected cells. The eight amino acid bleomycin hydrolase-like domain in the C-terminus, which does not contain any of the putative active site amino acids, was essential for protection against bleomycin induced chromatid breaks. These results demonstrate that intracellular hBH levels can influence the clastogenic action of bleomycin and that the C-terminus has a functional role in the biological activity of hBH. PMID- 9748473 TI - LAR tyrosine phosphatase receptor: proximal membrane alternative splicing is coordinated with regional expression and intraneuronal localization. AB - Examination of null-mutant Drosophila and Leukocyte Common Antigen-Related (LAR) deficient transgenic mice has demonstrated that the LAR protein tyrosine phosphatase (PTP) receptor promotes neurite outgrowth. In the absence of known ligands, the mechanisms by which LAR-type PTP receptors are regulated are unknown. We hypothesized that an alternatively spliced eleven amino acid proximal membrane segment of LAR (LAR alternatively spliced element-a; LASE-a) contributes to regulation of LAR function. Human, rat and mouse LAR cDNA sequences demonstrated that the predicted eleven amino acid inserts in rat and mouse are identical and share nine of eleven residues with the human insert. LASE-a splicing led to the introduction of a Ser residue into LAR at a position analogous to Ser residues undergoing regulated phosphorylation in other PTPs. In situ studies revealed increasingly region-specific expression of LASE-a containing LAR transcripts during postnatal development. RT-PCR analysis of cortical and hippocampal tissue confirmed that the proportion of LAR transcripts containing LASE-a decreases during development. Immunostaining of cultured PC12 cells, cerebellar granule neurons, dorsal root ganglia and sciatic nerve sections with antibody directed against the LASE-a insert demonstrated signal in cell bodies but little if any along neurites. In contrast, staining with antibody directed to a separate domain of LAR showed accumulation of LAR along neurites. The findings that LASE-a splicing is conserved across human, rat and mouse, that the LASE-a insert introduces a Ser at a site likely to be targeted for regulated phosphorylation and that developmentally regulated splicing is coordinated with specific regional and intraneuronal localization point to important novel potential mechanisms regulating LAR-type tyrosine phosphatase receptor function in the nervous system. PMID- 9748475 TI - DNA damage evaluated by the alkaline comet assay in lymphocytes of humans anaesthetized with isoflurane. AB - In the present paper, we report data on the possible DNA damage, induced in vivo by isoflurane using the alkaline single cell gel electrophoresis technique (SCGE comet assay) in patients before/after anaesthesia and in control group. Twelve patients, aged 22-66 years old, were anaesthetized for elective abdominal surgery with isoflurane in oxygen for 120-162 min (mean: 133.2 min). Venous blood samples were obtained from the patients before the induction of anaesthesia, at 60 and 120 min of anaesthesia and on the first, third and fifth following days of anaesthesia. SCGE was examined in 100 cells from each specimen graded as undamaged, intermediate and tailed nuclei. The number of undamaged nucleus was almost same in control and in patients before anaesthesia. However, significant differences were observed in proportion of undamaged, intermediate and tailed nucleus of patients at 60 and 120 min of anaesthesia and on the first day. DNA damage started to return to normal rates after the third day of anaesthesia and were almost identical with the rates of control group five days later. PMID- 9748476 TI - Structure of DNA topoisomerases. AB - Over the last several years topoisomerases have finally begun to yield to high resolution structural studies. These models have greatly aided our understanding of the mechanisms of topoisomerase catalysis and drug interactions. This review will cover advances in the structural biology of topoisomerases and discuss their implications for topoisomerase function. PMID- 9748477 TI - Cytogenetic adaptive response in cultured human lymphocytes: dependence on the time of exposure to adapting and challenging doses of gamma-rays. AB - Human lymphocytes from 16 healthy donors were exposed in vitro to an adapting dose of gamma-rays (0.05 Gy) at G0, or G1, or G1/S stage of the cell cycle and subsequently to a challenging dose of gamma-rays at G1, or G1/S, or S (1 Gy), or G2 (0.5 Gy) stage. Frequencies and distributions of the induced chromosome aberrations were analyzed in first-division metaphases. The data averaged over the donors revealed the protective action of the adapting exposure under the irradiation schemes with the challenging dose delivered at S or G2 stage. The majority of aberrations induced at these stages belonged to the chromatid type, and their yield was significantly higher in G2-exposed cells than in S-exposed cells. However, the relative reduction of the challenging dose effect (about 34%) in the adapted cells did not depend on the magnitude of this effect, and its value remained the same (within the experimental error) if aberrations were subdivided into chromosome and chromatid types or grouped as total deletions and total fragments. The adaptive response was not revealed under the schemes with the challenging dose delivered at G1 or G1/S stage. Analysis of the individual results showed that, in one and the same donor, the adaptive response could be observed under one irradiation scheme and not observed under other schemes, the most effective schemes being those with the challenging dose delivered at G2 stage. Four donors, however, did not show the adaptive response even under such schemes. Data on aberration distributions suggested that different repair processes, rather than a unique one, may underlie the adaptive response. PMID- 9748478 TI - Desensitization of delta-opioid-induced mobilization of Ca2+ stores in NG108-15 cells. AB - Activation of delta-opioid receptors in NG108-15 cells induces the release of calcium from an inositol 1,4,5-trisphosphate- sensitive intracellular store. We used fura-2-based digital imaging to study the effects of prolonged exposure to agonist on opioid-induced increases in [Ca2+]i. Exposure to D-Ala2-E-Leu5 enkephalin (DADLE) (1 microM) for 30 min completely desensitized NG108-15 cells to a second DADLE-induced response. The cells recovered gradually over 25 min following washout of DADLE. The desensitization was not due to depletion of intracellular calcium stores and bradykinin failed to cross-desensitize the DADLE evoked response, although both agonists mobilized the same Ca2+ store. Desensitization induced by 100 nM DADLE was overcome by a higher concentration of DADLE (100 microM). Treatment with 8-cpt-cAMP (0.1 mM) for 30 min did not influence the DADLE-induced increases in [CA2+]i. Phorbol dibutyrate (PdBu) (1 microM) blocked the response completely. Treatment with the inhibitor of cyclic nucleotide-dependent kinases H8 (1 microM) for 45 min did not prevent DADLE induced desensitization. Treatment with the protein kinase C (PKC) inhibitors staurosporin (10 nM) and GF-109203X (200 nM) for 45 min reduced desensitization. However, down-regulation of PKC by 24 h exposure to PdBu (1 microM) failed to prevent the DADLE-induced desensitization in NG108-15 cells. Thus, we conclude that multiple pathways participated in desensitization of delta-receptor-mediated Ca2+ mobilization, one of which includes PKC. PMID- 9748479 TI - Genetic analysis of adenovirus E1A: induction of genetic instability and altered cell morphologic and growth characteristics are segregatable functions. AB - Single multifunctional oncoproteins contribute to genomic instability development, but relationships between one or more oncoprotein-associated activities and genetic changes accompanying tumor cell progression are uncertain. Using NIH 3T3 derivative EN/NIH 2-20 containing transcriptionally silent neomycin phosphotransferase gene (neo) integrants with undetectable spontaneous reactivations, we studied wild-type (WT) and mutant adenovirus E1A-induced neo reactivation by neo-allelic rearrangement. WT E1A expression, yielding differential splice transcripts 12S and 13S and resulting in altered cell morphologic and growth characteristics, produced neo reactivations in 9 of 21 subclones (median rate per cell, 35 x 10(-6); range, 0.33 x 10(-6) to 936 x 10( 6)). Only 3 of 17 cell lines expressing CTdl976, a '12S' functional equivalent inducing altered cell morphologic and growth characteristics while lacking the 13S trans activation domain, yielded neo reactivations (range, 0.33 x 10(-6) to 0.67 x 10(-6)). One of 21 subclones expressing NTdl646, an E1A mutant retaining the trans domain but lacking p300 binding activity and the ability to alter cell morphologic and growth characteristics, produced neo reactivations (8.7 x 10( 6)). Other E1A mutants, all lacking the ability to alter cell morphologic and growth characteristics while binding pRb but variously lacking the trans domain and binding for p107 and/or p300, displayed undetectable neo-reactivations. 98 EN/NIH 2-20 derivatives coexpressing complementary mutant E1As exhibited altered morphologic and growth features, but only 10 of these produced neo reactivations, and maximum rates (14 x 10(-6)) were substantially lower than those in comparably derived, morphologically altered E1AWT-expressing counterparts (497 x 10(-6)). These findings suggest that maximum rates of gene reactivations by genomic rearrangement require the collective activities of functional domains assembled in single multifunctional proteins (or complexes) while altered cell morphologic and growth characteristics may arise through comparable sets of functional domains distributed across more than one protein (or complex). PMID- 9748480 TI - Testosterone differentially regulates expression of GnRH messenger RNAs in the terminal nerve, preoptic and midbrain of male tilapia. AB - The purpose of the present study was to examine the regulation of three molecular variants of gonadotropin-releasing hormone (GnRH)-encoding mRNAs by testosterone in the male tilapia Oreochromis niloticus. Tilapias castrated for two weeks were injected intraperitoneally with sesame oil or 5 microgram/g testosterone for 7 days. In situ hybridization histochemistry was performed using 35S-labelled 30 mer antisense oligonucleotide probes complementary to exon two (bases 1-30) of salmon-, seabream-, and chicken II-GnRH. Computerized image analysis was performed to quantify GnRH mRNA expression in the terminal nerve ganglia (nucleus olfactoretinalis) and in individual cells of the preoptic area and the midbrain tegmentum. Testosterone treatment significantly elevated terminal nerve salmon GnRH mRNA, reduced preoptic seabream-GnRH mRNA but had no effect on midbrain chicken II-GnRH mRNA levels. The total number and size of preoptic and midbrain GnRH mRNA-containing neurons or the total volume of the terminal nerve ganglia in testosterone-treated animals did not differ significantly from oil-treated animals. The midbrain chicken II-GnRH neurons are not targets of testosterone. These results demonstrate for the first time differential regulation of subpopulations of GnRH neurons with molecular diversity and different topography. PMID- 9748481 TI - Properties of the caspases. AB - Caspases comprise a structurally related group of cysteine proteases that share a dominant primary specificity for cleaving peptide bonds following Asp residues. Present in the cytosol of all animals, the caspases participate in proteolytic pathways required for executing programmed cell death, or apoptosis. In mammals the caspases have also evolved a function in activating proinflammatory cytokines. We review the current knowledge of the substrate specificity, structure, and activation mechanisms of human caspases and relate these findings to their fundamental biologic role. PMID- 9748482 TI - Bacterial and archeal type I topoisomerases. AB - Bacterial and archeal type I topoisomerases, including topoisomerase I, topoisomerase III and reverse gyrase, have different potential roles in the control of DNA topology including regulation of supercoiling and maintenance of genetic stability. Analysis of their coding sequences in different organisms shows that they belong to the type IA family of DNA topoisomerases, but there is variability in organization of various enzymatic domains necessary for topoisomerase activity. The torus-like structure of the conserved transesterification domain with the active site tyrosine for DNA cleavage/rejoining suggests steps of enzyme conformational change driven by DNA substrate and Mg(II) cofactor binding, that are required for catalysis of change in DNA linking number. PMID- 9748483 TI - A comparison of delta 9-THC and anandamide induced c-fos expression in the rat forebrain. AB - Rats were injected with the cannabinoid receptor agonists delta 9-THC (5 mg/kg) or anandamide (20 mg/kg) and assessed for changes in body temperature and locomotor activity. Their brains were then examined for the expression of the immediate early gene c-fos. Similar reductions in body temperature and locomotor activity were seen with delta 9-THC and anandamide although there was evidence, in line with previous reports, to suggest a shorter duration of action of anandamide. delta 9-THC and anandamide caused equally high levels of c-fos expression in the paraventricular nucleus of the hypothalamus and the lateral septum. Both drugs also increased c-fos expression in the central nucleus of the amygdala although the effect was greater with delta 9-THC. Only delta 9-THC caused significant increases in c-fos expression in the nucleus accumbens and caudate-putamen. These differences may be linked to differential activation of cannabinoid receptor subtypes or to differences in efficacy in activating second messenger systems linked to cannabinoid receptors. These findings complement evidence of qualitative differences in the actions of anandamide and delta 9-THC emerging from tests of drug discrimination, cross-tolerance, conditioned place preference and anxiety. PMID- 9748484 TI - Biphasic changes in the levels of N-methyl-D-aspartate receptor-2 subunits correlate with the induction and persistence of long-term potentiation. AB - N-Methyl-D-aspartate glutamate receptors (NMDAR) form ion channels made up of polypeptides from two classes of subunits; NR1 is obligatory for function whereas members of the NR2 class regulate the properties of the channel. Long-term potentiation (LTP) of synaptic transmission is an event largely dependent on NMDAR activation, and is studied as the primary cellular model of memory in the mammalian brain. While there has been a focus on non-NMDARs in mediating the expression of LTP, we report here biochemical evidence for plasticity of the NMDAR that is associated with LTP persistence in awake animals. Following the establishment of LTP in perforant path synapses of the dentate gyrus, we observed a rise in NR2B protein levels 48 h post-tetanus which was dependent upon activation of NMDARs during the tetanization, and which strongly correlated with the degree of LTP measured at this time-point. We also observed a transient increase in both NR2B and NR2A protein levels 20 min post-tetanus that returned to control levels by 4 h. These early increases were not observed in anaesthetized animals which do not sustain persistent LTP. Our data demonstrate a marked plasticity of NMDAR subunit expression, which may affect LTP persistence, as well as the subsequent ability to induce LTP at previously activated synapses. PMID- 9748485 TI - Comparison of ethylene, propylene and styrene 7,8-oxide in vitro adduct formation on N-terminal valine in human haemoglobin and on N-7-guanine in human DNA. AB - Epoxides react at various nucleophilic sites in macromolecules such as haemoglobin and DNA. To study the reaction rate constants of ethylene oxide (EO), propylene oxide (PO) and styrene 7,8-oxide (SO) towards two of these positions, i.e., the N-terminal valine in haemoglobin and N-7-guanine in DNA was the central aim of this investigation. These two reactive sites are the most studied haemoglobin and DNA adducts, respectively. Further attention, therefore, was also paid to the applicability in vivo of the in vitro determined reaction constants. The determination of the second-order rate constants between EO and PO and N terminal valine in Hb [2.7 l (mol Hb h)-1 and 1.0 l (mol Hb h)-1, respectively] were consistent with the literature values. The constants for the reaction with N 7-guanine [16x10(-3) l (mol DNA nucleotide h)-1 and 7. 7x10(-3) l (mol DNA nucleotide h)-1, respectively] were lower than previously published values, probably due to differences in the methodology used. The use of the in vitro obtained values to model the in vivo situation lead to a consistent picture for EO and PO. In contrast, for SO the in vitro ratio between the adduct formation on N-terminal valine [1.5 l (mol Hb h)-1] and N-7-guanine [0.71x10(-3) l (mol DNA nucleotide h)-1] was about two orders of magnitude greater than for the in vivo situation. This was probably due to a lower than expected reactivity of SO towards N-terminal valine in vivo. Further research is needed to elucidate whether the use of SO in vitro, contrasting with the in vivo experiments in which SO was metabolically formed from styrene, could entail an explanation for this discrepancy. Concerning the methodological part, the use of dipeptide standards to replace the alkylated globins as standard lead to an improvement of the method. Especially the commercial availability of the standards, their stability and accurately known adduct content will make them to the standards of choice in the future. PMID- 9748486 TI - An age-associated correlation between cellular bioenergy decline and mtDNA rearrangements in human skeletal muscle. AB - Post-mitotic tissues such as skeletal muscle develop a tissue bioenergy mosaic during the process of normal aging that eventually culminates into a bioenergetically diverse tissue containing cells ranging in their oxidative phosphorylation capacity from normal to grossly defective. The mosaic is postulated to develop continuously from birth with the relative proportions of cytochrome c oxidase (COX) proficient (positive) and COX deficient (negative) muscle fibers differing dramatically as a function of age. Generally, young individuals only display the rare fiber deficient in COX activity while aged individuals show a significantly higher proportion of negative fibers. There appears to be a random element governing which cells will be affected. Consequently, adjacent cells within a given tissue may exhibit vastly differing COX activities. Multiple mitochondrial DNA (mtDNA) deletions also appear to accumulate in skeletal muscle, similarly displaying a dramatic disparity as a function of age. Our previous findings have indicated that the accumulation of multiple mtDNA deletions, along with a concurrent decrease in wild-type mtDNA, strongly correlates with the age-associated decrease in COX activity observed in skeletal muscle. Although no definitive associations were established at the cellular level, an important prediction arose from this study. Cells that accumulate large numbers of mitochondrial mutations and have reduced levels of full-length mtDNA would be expected to be severely affected and show reduced COX activity as a consequence. Cells that accumulate fewer mutations or retain adequate amounts of wild-type mtDNA would be predicted to be less affected or even retain normal oxidative metabolism. In order to establish a link associating COX activity to the status of mtDNA within individual fibers, we developed single cell extra-long PCR (XL-PCR). The procedure was used to assess the relative concentration of full-length mtDNA with respect to any mtDNA deletions detected in individual human skeletal muscle fibers of 'pre-established' COX activity. Single cell XL-PCR analysis of COX positive fibers dissected from a 5-year old and 90-year old individual showed that 80% or more of the fibers contained full length mtDNA and few, if any, mtDNA rearrangements. COX deficient or COX intermediate fibers taken from the same individuals, by contrast, depicted a heterogeneous population of rearranged mtDNA species with no detectable full length mtDNA. The data presented here indicates that COX deficient muscle fibers extracted from individuals, regardless of age, were accompanied by extensive mtDNA rearrangements and reduced levels of full-length mtDNA. This provides compelling evidence linking mtDNA mutations to COX activity decline in skeletal muscle and has important implications when considering the molecular basis of the aging process. PMID- 9748487 TI - Diencephalic and mesencephalic projections to rhombencephalic reticular nuclei in lampreys. AB - Behavioral studies in lampreys of the northern genera, Ichthyomyzon, reveal that sensory inputs initiate and modulate locomotion by activation of reticulospinal (RS) neurones, which constitute the primary descending system involved in motor activity. The interneurones relaying afferent vestibular, trigeminal, lateral line, cutaneous and proprioceptive inputs are localized in the rhombencephalic region of the lamprey brainstem, unlike the visual inputs that are relayed in the mesencephalic region. The knowledge of diencephalic-mesencephalic cell distributions that project to the RS neurones is limited. They were isolated by iontophoretically injecting cobalt-lysine in vitro into the middle (MRRN) and posterior (PRRN) rhombencephalic reticular nuclei of Petromyzon marinus and Ichthyomyzon unicuspis, Fourteen of 31 injections were successful (MRRN, 7; PRRN, 7). Cell groups were labeled ipsilateral to the injection site in the thalamus (corpi geniculati; pars dorsalis thalami lateralis and medialis; nucleus (n.) subhabenularis lateralis), in the epithalamus (n. commissura posteriori) and in the pretectum. Cell groups were labeled bilaterally within the dorsal region along the diencephalic-mesencephalic border (caudal pretectum and rostral tectum opticum), in tectum opticum, torus semicircularis, and tegmentum mesencephali. There were more backfilled cells from MRRN injections (538-6466 cells) than from PRRN injections (53-553 cells) (MW Rank Sum, p < 0.001). The cell bodies were less than 40 microns long ipsilateral to the injection site, and longer contralaterally. Those greater than 50 microns were backfilled from PRRN injections. The location and organization of the cell groups identified is comparable to that of other vertebrates. PMID- 9748489 TI - DNA gyrase and topoisomerase IV: biochemical activities, physiological roles during chromosome replication, and drug sensitivities. AB - DNA gyrase and topoisomerase IV are the two type II topoisomerases present in bacteria. Though clearly related, based on amino acid sequence similarity, they each play crucial, but distinct, roles in the cell. Gyrase is involved primarily in supporting nascent chain elongation during replication of the chromosome, whereas topoisomerase IV separates the topologically linked daughter chromosomes during the terminal stage of DNA replication. These different roles can be attributed to differences in the biochemical properties of the two enzymes. The biochemical activities, physiological roles, and drug sensitivities of the enzymes are reviewed. PMID- 9748488 TI - Neurotrophic action of lipocortin 1 derived from astrocytes on cultured rat cortical neurons. AB - The lipocortins are a family of structurally related proteins, namely an annexin family, that exerts a variety of cellular functions through Ca2+-dependent binding to phospholipase A2 [EC 3.1. 1.4], including a crucial role in the central nervous system (CNS) such as antipyrogenic, thermoregulatory and neuroprotective agents in vivo. To elucidate the paradigm of lipocortin 1 functions in the CNS, we have first demonstrated (1) the induction and subsequent extracellular secretion of LC1 by glucocorticoid in cultured rat astrocytes, and (2) neurotrophic activities (survival-promoting, neuritogenic and synaptogenic actions on rat cortical neurons) of recombinant LC1. Time-and dose-dependent experiments of a synthetic glucocorticoid, dexamethasone (DEX), on rat cortical astrocytes in culture revealed that the expression of the intracellular LC1 mRNA and protein were significantly augmented by DEX (1 microM). In addition, DEX evoked an extracellular secretion of LC1 without its cytotoxic effects. Furthermore, the recombinant LC1 appeared to promote not only the survival and neurite outgrowth but also the synaptogenesis of embryonal rat cortical neurons. These results suggest that LC1 induced and selectively released from astrocytes by either endogenously or exogenously introduced glucocorticoids may play a specific and essential role on development and regeneration of the central nervous system. PMID- 9748490 TI - Rat liver phosphoribosylpyrophosphate synthetase is activated by free Mg2+ in a manner that overcomes its inhibition by nucleotides. AB - Phosphoribosylpyrophosphate synthetase is activated by Pi and free Mg2+ as an essential activator and inhibited by nucleotides, especially ADP and GDP. The rat liver enzyme is a complex aggregate of two highly homologous catalytic subunits (PRS I and PRS II) and two associated proteins (PAP39 and PAP41). PRS I is more sensitive to inhibition by ADP and GDP than is PRS II. The native liver enzyme showed a weaker sensitivity to inhibition by nucleotides than expected from its composition. To further understand the regulation of the liver enzyme, kinetic studies of each subunit component and the liver enzyme regarding Mg2+ activation and inhibition by ADP and GDP were carried out. Assay conditions were designed to keep free Mg2+ at constant concentrations. (1) GDP, as MgGDP, did not affect the apparent Km values of PRS I for MgATP and ribose-5-phosphate but did dramatically increase the apparent Ka value for free Mg2+. (2) In contrast, ADP, as MgADP, increased the Km value for MgATP of PRS I as well as the Ka value for free Mg2+. (3) High concentrations of free Mg2+ almost completely nullified the inhibitory effect of MgGDP and partly that of MgADP on PRS I. (4) At low free Mg2+ concentrations within the physiological range, inhibition by the nucleotides is of physiological significance and conversely, variation in free Mg2+ concentrations critically affects the enzyme activity in the presence of inhibitory nucleotides. (5) The response of PRS II and the native liver enzyme is similar to that of PRS I, while the effects of MgGDP and MgADP were smaller than that on PRS I. (6) We propose that MgGDP binds to a regulatory site of PRS I and PRS II and MgADP to the substrate MgATP site and also the regulatory site. The allosteric interaction of the regulatory site and the Mg2+ binding site is also considered. PMID- 9748491 TI - Effects of antioxidants on streptozotocin-induced clastogenesis in mammalian and insect cells. AB - The effects of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT) and the hydroxyl radical scavenger mannitol (MAN) on the clastogenesis induced by STZ in Chinese hamster ovary (CHO) and mosquito cells were investigated. The addition of liposome-entrapped SOD, CAT and MAN to both cell lines caused a significant decrease in the yield of STZ-induced chromosome aberrations (p<0.05). However, the inhibitory effect was more evident in CHO (40.6-67.5%) than in mosquito (15.2-53.6%) cells. These findings indicate that the chromosome damage induced by STZ can be partially inhibited through the incorporation of antioxidant compounds into the cells and suggest that free radicals are involved in the clastogenesis by STZ. PMID- 9748492 TI - Genetic damage induced by methylglyoxal and methylglyoxal plus X-rays in Drosophila melanogaster germinal cells. AB - The effect of methylglyoxal (MG) and MG administered prior to X-irradiation was investigated in Drosophila melanogaster germinal cells using the sex-linked recessive lethal (s.l.r.l.), II-III autosomal translocation (AT) and X-chromosome nondisjunction (ND) tests. For the s.l.r.l. test the males were either injected with MG (0.5 M, 0.75 M or 1.7 M) or fed for 24 h (1 M) and two 24 h broods (A and B) were obtained. For the AT test the males were injected with MG 1.7 M and the same brooding scheme was followed. ND was tested in females fed on MG 1 M. The only effect observed after MG treatment was a significant increase on the yield of s.l.r.l. with MG 1.7 M. In the combined treatments MG was administered prior to irradiation with 20 Gy of X-rays and the induction of s.l.r.l. and AT was assessed. Pre-treatment with MG 0.75 M and 1.7 M enhanced the frequency of s.l.r.l. in cells sampled in brood B, consisting mainly of the rather hypoxic late spermatids. It is suggested that this radiosensitizing effect could be ascribed to a decrease in the level of glutathione due to the metabolization of MG. PMID- 9748493 TI - Upregulation of amyloid precursor protein gene promoter in rat primary hippocampal neurons by phorbol ester, IL-1 and retinoic acid, but not by reactive oxygen species. AB - The APP gene promoter has multiple regulatory sequences, some of which may contribute to the neuropathology of Alzheimer's disease (AD). In this study, we investigated the effects of phorbol ester (PMA), IL-1, retinoic acid and reactive oxygen species on APP promoter activity in primary hippocampal neurons. We transfected neurons with either of two APP promoter constructs, a -2.8 kb and a shorter -488 bp upstream fragment fused to the chloramphenicol transferase (CAT) reporter gene. We demonstrated that phorbol 12-myristate-13 acetate (PMA), retinoic acid and IL-1 all stimulated both APP promoter constructs in hippocampal neurons after 24 h treatment. PMA and IL-1 treatments led to 2-fold increases of APP promoter activity. Retinoic acid induced a 3-fold increase. In addition, the magnitude of APP promoter responses to PMA and IL-1 treatment was similar between APP -2.8 kb and -488 bp plasmid transfected neurons. This suggests that the AP-1 sequence at -350 to -344 in the APP promoter may mediate the stimulatory effects of PMA and IL-1, as previously observed in endothelial and HeLa cells. In contrast, hydrogen peroxide, which was shown to activate NF-kappaB in primary neurons, failed to stimulate APP promoter activity, suggesting that the regulatory elements in the APP promoter may not respond to reactive oxygen species. Overall, these data indicate that APP expression in primary neurons can be modulated by PMA, IL-1 and retinoic acid. However, the contribution of reactive oxygen to Alzheimer's disease may not be directly related to the activation of the APP gene promoter but instead to neuronal damage associated with oxidative stress. Since elevated levels of IL-1 have been observed in AD brain, IL-1 could contribute to development of Alzheimer's disease by stimulating APP synthesis in primary neurons. PMID- 9748494 TI - Butyrylcholinesterase-catalysed hydrolysis of aspirin, a negatively charged ester, and aspirin-related neutral esters. AB - Although aspirin (acetylsalicylic acid) is negatively charged, it is hydrolysed by butyrylcholinesterase (BuChE). Catalytic parameters were determined in 100 mM Tris buffer, pH 7.4, in the presence and absence of metal cations. The presence of Ca2+ or Mg2+ (<100 mM) in buffer did not change the Km, but accelerated the rate of hydrolysis of aspirin by wild-type or D70G mutant BuChE by 5-fold. Turnover numbers were of the order of 5000-12000 min-1 for the wild-type enzyme and the D70G and D70K enzymes in 100 mM Tris, pH 7.4, containing 50 mM CaCl2 at 25 degreesC; Km values were 6 mM for wild-type, 16 mM for D70G and 38 mM for D70K. People with 'atypical' BuChE have the D70G mutation. The apparent inhibition seen at high aspirin concentration was not due to inhibition by excess substrate but to spontaneous hydrolysis of aspirin, causing inhibition by salicylate. The wild-type and D70G enzymes were competitively inhibited by salicylic acid; the D70K enzyme showed a complex parabolic inhibition, suggesting multiple binding. The effect of salicylate was substrate-dependent, the D70K mutant being activated by salicylate with butyrylthiocholine as substrate. Km value for wild-type enzyme was lower than for D70 mutants, suggesting that residue 70 located at the rim of the active site gorge was not the major site for the initial encounter aspirin-BuChE complex. On the other hand, the virtual absence of affinity of the W82A mutant for aspirin indicated that W82 was the major residue involved in formation of the Michaelis complex. Molecular modelling of aspirin binding to BuChE indicated perpendicular interactions between the aromatic rings of W82 and aspirin. Kinetic study of BuChE-catalysed hydrolysis of different acetyl esters showed that the rate limiting step was acetylation. The bimolecular rate constants for hydrolysis of aspirin by wild-type, D70G and D70K enzymes were found to be close to 1x106 M-1 min-1. These results support the contention that the electrostatic steering due to the negative electrostatic field of the enzyme plays a role in substrate binding, but plays no role in the catalytic steps, i.e. in the enzyme acetylation. PMID- 9748495 TI - Bleomycin sensitivity test in the exposed and reference human populations. AB - Sensitivity to bleomycin was investigated in lymphocytes collected from three groups of males: 30 occupationally exposed cokery workers, 38 environmentally exposed Silesian citizen and 35 rural inhabitants. The data were analyzed at both the individual and group levels. The first analysis has revealed a substantial interindividual variability in the level of generated breaks (breaks per cell, b/c). This variability was independent of the age of the donor, smoking habit and X-ray exposure as tested in the multiple regression model. The means per group for the occupationally and environmentally exposed persons were almost the same with the values of 0.674 and 0.639, respectively. These two groups differed significantly from the rural population (b/c=0.448, p<0.001 by MANOVA). The reproducibility of the assay was satisfying (p>0.49 by the Wilcoxon matched paired test) after omitting 7 out of 49 repeatedly sampled donors. Those persons exhibited extremely high b/c rates in the first sampling. PMID- 9748496 TI - Clinical applications of quinolones. AB - The quinolone antimicrobials are the class of inhibitors of bacterial topoisomerases that has been developed most fully for clinical use in human medicine. Initial members of the class had their greatest potency against Gram negative bacteria, but newly developed members have exhibited increased potency against Gram-positive bacteria and soon agents will be available with additional activity against anaerobic bacteria, providing a broad spectrum of potency. After nalidixic acid, the earliest member of the class which was used for treatment of urinary tract infections, the later fluoroquinolone congeners have had sufficient potency, absorption, and distribution into tissue for additional uses in treatment of sexually transmitted diseases, infections of the gastrointestinal tract, respiratory tract, skin, and bones and joints. Tolerability of these agents in usual doses has been good. Acquired bacterial resistance resulting from clinical uses has occurred in particular among staphylococci and Pseudomonas aeruginosa. Intense drug use and ability of resistant pathogens to spread have also contributed to development of resistance in initially more susceptible pathogens such as Escherichia coli and Neisseria gonorrhoeae in certain settings. Preservation of the considerable clinical utility of the quinolone class for the long term will be affected by the extent to which their use is judicious. PMID- 9748497 TI - Radiation-induced long-lived radicals which cause mutation and transformation. AB - Using electronic spin resonance (ESR), we found a new type of radical with a long life-time in cells (T1/2>20 h) and which may play a more important role in the induction of mutation and transformation than either the active, short-lived, H, or OH radicals. When cells were treated with dimethyl sulfoxide (DMSO) and l ascorbic acid (AsA) just before irradiation, the short-lived radicals were well scavenged. On the other hand, if cells were treated with the scavengers 20 min after irradiation, then AsA scavenged the long-lived radicals, but DMSO did not. AsA treatment 20 min after the start of irradiation drastically reduced both the frequencies of mutation at the hypoxanthine guanine phosphoribosyl transferase (HGPRT) locus in human cells and morphological transformations in mouse m5S cells, but DMSO treatment did not. In addition, AsA treatment 20 h after irradiation also reduced the mutation frequency in human cells. These results suggested that mutations and morphological transformation are probably caused by the presence of long-lived radicals in the cells, rather than by short lived radicals, and that AsA reacts efficiently with long-lived radicals, resulting in a decrease of the mutations and transformations induced. PMID- 9748498 TI - The influence of DNA ploidy of a human tumor cell line on the frequencies of micronuclei or chromosome aberrations after irradiation. AB - To develop methods for assessing the intrinsic cellular radiosensitivity, it is important to evaluate the relationship between DNA ploidy of cells and frequencies of micronuclei (MN) or chromosome aberrations after irradiation. From the original human fibrosarcoma cell line HT-1080, we isolated two clones which have different chromosome ploidy: clone 5 is pseudodiploid and clone 1 is heteroploid. We examined the radiosensitivity of the two clones using a clonogenic cell survival assay and a cytokinesis-block MN assay, and by scoring chromosome aberrations using the premature chromosome condensation (PCC) method combined with a fluorescence in situ hybridization (FISH) procedure immediately and at 24 h after irradiation. The MN frequency increased according to the irradiation dose in both clones. The MN frequency of clone 1 was significantly higher than that of clone 5 regardless of whether the assay was performed immediately or 24 h after irradiation. However, when the numbers of MN were normalized by the DNA index of each clone, a significant difference in the frequency of MN was not observed. In the PCC and FISH studies, there was a linear relationship between the radiation dose and the initial breaks of chromosome 4, but the breaks of clone 1 were much more frequent than those of clone 5. Twenty four h after irradiation, the chromosome 4 breaks of clone 1 were observed much more frequently than those of clone 5 at the same radiation dose. When the numbers of chromosome 4 breaks were normalized by the number of chromosome 4 in each clone without radiation, no such difference in the number of breaks was observed. These findings demonstrated that the DNA content or chromosome ploidy influenced the induction of the MN or chromosome aberrations in HT-1080 cells after irradiation. PMID- 9748499 TI - Synaptic activity-dependent modulation of mitochondrial gene expression in the rat hippocampus. AB - In order to identify genes that may underlie the maintenance of long-term potentiation (LTP) at perforant path synapses, complementary DNA libraries were synthesised from dentate gyrus total RNA extracts prepared 48 h after the induction of LTP and from control dentate gyrus extracts. Through differential screening of the LTP library we have identified the mitochondrial 12S rRNA (mt12SrRNA) as a transcript that was elevated at this late time. Northern blot analyses showed that the elevation in mt12SrRNA expression began around 8 h and persisted for at least 2 weeks post-tetanus. We then examined the expression patterns of other mitochondrially-encoded genes and demonstrated a similar elevation in their expression. mt12SrRNA levels were also elevated in other hippocampal regions, including areas CA3 and CA1 and were elevated following low frequency stimulation or in the presence of an N-methyl-D-aspartate receptor antagonist where induction of LTP was precluded. Taken together, these observations suggest that a long-lasting up-regulation of energy production may be triggered by synaptic activity and this activity need not be of sufficient strength to induce LTP, but may be related to the induction of a metaplastic state. PMID- 9748500 TI - Resistance of butyrylcholinesterase to inactivation by ultrasound: effects of ultrasound on catalytic activity and subunit association. AB - The effects of 20 kHz ultrasound on catalytic activity and structure of the tetramer of wild-type human butyrylcholinesterase (BChE) from plasma and recombinant D70G mutant enzyme were studied at constant temperature. Effects on catalytic properties of both enzymes were investigated by kinetic analysis under ultrasound irradiation using a neutral substrate (o-nitrophenylbutyrate), a positively charged substrate (butyrylthiocholine), and a negatively charged substrate (aspirin). Effects on structure of highly purified wild-type BChE were followed by gel electrophoresis and activity measurements at Vmax after ultrasound treatment. Unlike hydrostatic pressure, mild ultrasound had moderate effects on catalytic parameters of BChE-catalyzed hydrolysis of substrates. For both wild-type and D70G, Km increased slightly with butyrylthiocholine and o nitrophenylbutyrate under ultrasound irradiation, suggesting that these effects of ultrasound were not due to the periodic variation of pressure but rather to shear forces that took off substrate from the peripheral site and altered diffusion to the active site. By contrast, affinity of the D70G mutant for aspirin slightly increased with ultrasound power, suggesting that ultrasound induced microstreaming unmasked peripheral residues involved in recognition and initial binding of the negatively charged substrate. Results support the contention that Km is a composite affinity constant, including dissociation constant of the first encounter enzyme-substrate complex on the peripheral site. Small changes in catalytic activity may have resulted from ultrasound-induced subtle conformational changes altering the active site reactivity. Short ultrasound irradiation induced a faint transient enzyme activation, but prolonged irradiation caused partial dissociation of the tetrameric enzyme and irreversible inactivation. Partial dissociation was related to enzyme microheterogeneity, i.e., nicked (C-terminal segment depleted) tetramers were less stable than native tetramers. The resistance of the native tetramer to ultrasound-induced dissociation was ascribed to the existence of an aromatic amino acid array on the apolar side of the C-terminal helical segment of subunits, the four subunits being held together in a four-helix bundle containing the aromatic zipper motifs. Aromatic/aromatic interactions between the four helical segments are thought to be enhanced by ultrasound-generated pressure. PMID- 9748501 TI - Pharmacological evidence for GABAergic and glutamatergic involvement in the convulsant and behavioral effects of glutaric acid. AB - The effect of intrastriatal administration of glutaric acid (GTR), a metabolite that accumulates in glutaric acidemia type I (GA-I), on the behavior of adult male rats was investigated. After cannula placing, rats received unilateral intrastriatal injections of GTR buffered to pH 7.4 with NaOH or NaCl. GTR induced rotational behavior toward the contralateral side of injection and clonic convulsions in a dose-dependent manner. Rotational behavior was prevented by intrastriatal preadministration of DNQX and muscimol, but not by the preadministration of MK-801. Convulsions were prevented by intrastriatal preinjection of muscimol. This study provides evidence for a participation of glutamatergic non-NMDA and GABAergic mechanisms in the GTR-induced behavioral alterations. These findings may be of value in understanding the physiopathology of the neurological dysfunction in glutaric acidemia. PMID- 9748502 TI - Influence of vindesine exposure on the micronucleus formation and cell survival in V79 cells. AB - Effect of different concentrations (0, 1, 5, 10, 20 and 50 nM) of vindesine sulphate was studied on clonogenicity and micronucleus (MN) formation in V79 (Chinese hamster lung fibroblasts) cells. Exposure of V79 cells to vindesine for 6 h resulted in a concentration dependent decline in cell survival. The frequency of micronuclei (MN) increased in a concentration dependent manner at 16, 22 and 28 h post-exposure. The frequency of MN increased significantly after 5 to 50 nM drug exposure at 16 and 22 h post-treatment, while increasing post-exposure time to 28 h resulted in a significant increase in MN frequency at all exposure doses of vindesine. The statistical evaluation of concurrent concentrations at various time periods showed a non-significant difference in MN frequency among various post-exposure time periods, except 16 h and 28 h for 50 nM, where a significant decline in the MN frequency was observed at 28 h compared to 16 h post-exposure. The cell proliferation indices showed a concentration dependent decline in the frequency of binucleate cells and this decline was linear quadratic. The increasing drug concentration resulted in a concentration dependent decline in cell survival. While the frequency of micronuclei increased the cell survival decreased and the relationship between cell survival and micronucleus induction was linear quadratic. PMID- 9748503 TI - Differential expression of SAPK isoforms in the rat brain. An in situ hybridisation study in the adult rat brain and during post-natal development. AB - MAPK pathways transduce a broad variety of extracellular signals into cellular responses. Despite their pleiotropic effects and their ubiquitous distribution, surprisingly little is known about their involvement in the communication network of nerve cells. As a first step to elucidate the role of MAPK pathways in neuronal signalling, we studied the distribution of SAPK alpha/JNK2, SAPK beta/JNK3, and SAPK gamma/JNK1, three isoforms of SAPK/JNK, a stress-activated MAPK subfamily. We compared the mRNA localisation of the three main isoforms in the adult and developing rat brain using in situ hybridisation. In the adult brain, SAPK alpha and beta were widely but heterogeneously distributed, reproducing the pattern of a probe that does not discriminate the isoforms. Differently, high labelling for the SAPK gamma probe was exclusively localised in the endopiriform nucleus and medial habenula. Intermediate staining was detected in the hippocampus. During post-natal development, SAPK beta showed the same localisation as in the adult. Nevertheless, the semi-quantitative analysis of optical densities showed significantly different mRNA levels. In the adult, SAPK gamma signal was weak, whereas in newborn rats the labelling was intense and widely distributed. SAPK gamma mRNA levels decreased during development, to reach the low signals detected in the adult. These results suggest that in the central nervous system SAPK-type MAP kinases perform significant physiological functions which are particularly relevant during post-natal development. The distinct distribution patterns of SAPK isoforms in the adult rat brain support the hypothesis that separate functions are performed by the products of the three SAPK genes. PMID- 9748504 TI - Corrigendum to: 'Mutagenicity of p-aminophenol in escherichia coli WP2 uvrA/pKM101 and its relevance to oxidative DNA damage' PMID- 9748505 TI - Autoradiographic visualisation of axonal transport of adenosine A1 receptors along the rat vagus nerve and characterisation of adenosine A1 receptor binding in the dorsal vagal complex of hypertensive and normotensive rats. AB - The present study had employed in vitro receptor autoradiography with [3H]DPCPX to visualise the presence of adenosine A1 receptors on the rat nodose ganglion, which contains the perikarya of vagal afferent neurons projecting the the nucleus tractus solitarius (NTS). In addition, unilateral vagal ligation resulted in an accumulation of [3H]DPCPX binding adjacent to the ligatures, indication that adenosine A1 receptors are subject to axoplasmic flow along the rat vagus nerve. Radioligand binding assays were utilised to characterise the properties of adenosine A1 receptors in the dorsal vagal complex (NTS, area postrema and dorsal motor nucleus of the vagus) of pup and adult normotensive (Wistar Kyoto, WKY) and hypertensive (spontaneously hypertensive, SHR) rats. Saturation binding indicated that the affinity (KD) of [3H]DPCPX, and the binding site density (Bmax) were not different between the adult WKY and SHR, although the pup SHR had a lower KD value than the pup WKY rat. Competition binding assays revealed complex differences between the two rat strains; however, with respect to hypertension, the affinity of the selective adenosine A1 agonist, cyclohexyladenosine (CHA), was markedly reduced in the membranes from SHR (Ki approximately 93 nM) compared to WKY (approximately 6 nM). Such an observation is consistent with the attenuated responses of SHRs to intra-NTS injections of adenosine. PMID- 9748506 TI - Investigating the biological functions of DNA topoisomerases in eukaryotic cells. AB - DNA topoisomerases participate in nearly all events relating to DNA metabolism including replication, transcription, and chromosome segregation. Recent studies in eukaryotic cells have led to the discovery of several novel topoisomerases, and to new questions concerning the roles of these enzymes in cellular processes. Gene knockout studies are helping to delineate the roles of topoisomerases in mammalian cells, just as similar studies in yeast established paradigms concerning the functions of topoisomerases in lower eukaryotes. The application of new technologies for identifying interacting proteins has connected the studies on topoisomerases to other areas of human biology including genome stability and aging. These studies highlight the importance of understanding how topoisomerases participate in the normal processes of transcription, DNA replication, and genome stability. PMID- 9748507 TI - Molecular cloning of the dnaK locus, and purification and characterization of a DnaK protein from Bacillus brevis HPD31. AB - Using part of the dnaK gene from Bacillus subtilis as a probe, a 4. 4-kbp SacI BglII fragment of chromosomal DNA of Bacillus brevis, a protein-hypersecreting bacterium, was cloned. Nucleotide sequencing revealed 3 open reading frames in the order of grpE-dnaK-dnaJ homologues. We purified DnaK protein to homogeneity from B. brevis HPD31 harboring a multi-copy dnaK expression plasmid. Purified DnaK showed ATPase activity which was synergistically stimulated 14-fold by the addition of glutathione S-transferase-DnaJ and glutathione S-transferase-GrpE fusion proteins. DnaK hydrolyzed not only ATP but also CTP, UTP, and GTP at about 40% of the efficiency of ATP. The specific activity of DnaK-ATPase was 7.25x10-3 unit/mg protein (the turnover number against ATP was 0.47 min-1) under our assay conditions. The DnaK dimers dissociated into monomers on addition of ATP, GTP, CTP, UTP and ATPgammaS, but not ADP or AMP. DnaK formed a stable complex with permanently unfolded carboxymethylated alpha-lactalbumin but not with native alpha-lactalbumin, and this complex was dissociated by addition of ATP/Mg. Formation of this complex was inhibited in the presence of inorganic phosphate. PMID- 9748508 TI - The heterocyclic amine binding receptors of Lactobacillus gasseri cells. AB - Lactobacillus gasseri is a common inhabitant of human intestine. The L. gasseri strains SBT10239 and SBT10241 have shown high antimutagenicity and binding properties with different heterocyclic amines. In order to identify the cell wall components involved in binding with the heterocyclic amines, the cells and cell walls of L. gasseri strains were subjected to different chemical and enzymatical treatments, prior to the binding experiments. The results indicated that the binding receptors for heterocyclic amines are the carbohydrate moieties of the cell wall. Binding of the heterocyclic amines with L. gasseri cell walls and the carbohydrate content showed high correlation coefficient, whereas it was insignificant or negative with protein content. The lectin binding studies revealed that the glucose molecules of the cell wall has a significant role in binding the heterocyclic amines. The inhibition caused by the lectin Concanavalin A was reversed when treated with methyl glucoside, a competitive inhibitor of Concanavalin A and restored the binding of heterocyclic amine with the cells. PMID- 9748509 TI - Rapid sodium channel augmentation in response to inflammation induced by complete Freund's adjuvant. AB - The mechanisms by which inflammation induces a chronic pain state are poorly understood. Following the induction of many painful conditions, an increase in the spontaneous firing rate of neurons is often observed in peripheral sensory ganglia. Since ion channels are essential mediators of spike generation and impulse conduction, it is reasonable to postulate that local changes in ion channel expression might underlie the changes in membrane excitability. Such alterations may serve to enhance the efficiency by which painful stimuli are transduced and then conducted to the central nervous system. In these studies, we employed immunocytochemical methods to investigate the changes in sodium channel expression in dorsal root ganglia of rats following a subcutaneous injection of complete Freund's adjuvant, an inducer of chronic inflammation. We find that sodium channel immunoreactivity within primary sensory neurons is dramatically increased within 24 h of the complete Freund's adjuvant injection. These changes persist in small neurons for at least 2 months and roughly parallel the time course of behaviorally measured changes in pain thresholds. Thus, the regulation of sodium channel synthesis may play a role in the generation and maintenance of the hyperesthetic state seen in chronic inflammation. PMID- 9748510 TI - Distribution of neuropeptide Y receptor expression in the rat suprachiasmatic nucleus. AB - Neurones of the suprachiasmatic nucleus constitute the mammalian circadian clock which receives photic information via the retino-hypothalamic tract and to some extent non-photic information via the geniculo-hypothalamic tract. The majority of neurones in the geniculo-hypothalamic tract contains neuropeptide Y and both in vitro and in vivo physiological experiments have demonstrated that neuropeptide Y administered directly into the suprachiasmatic nucleus has the capacity to phase-shift the endogenous circadian rhythm of these neurones. The recent cloning of multiple mammalian neuropeptide Y receptors enabled us to perform an in situ hybridization histochemical study identifying expression of distinct neuropeptide Y receptor encoding mRNAs in the suprachiasmatic nucleus. It was seen that Y1 and Y5 receptor mRNA is highly expressed in neurones of the ventrolateral portion of the suprachiasmatic nucleus while neither Y2 nor Y4 receptor mRNA could be detected in the nucleus. These experiments demonstrate that post-synaptic neuropeptide Y mediated events in the suprachiasmatic nucleus are likely to be mediated by either of these receptors. PMID- 9748511 TI - Generation of a genetic polymorphism in clonal populations of the bacterium Streptomyces ambofaciens: characterization of different mutator states. AB - In Streptomyces ambofaciens, colony pigmentation is an unstable character. Very unstable mutants selected from twelve wild type (WT) subclones of S. ambofaciens ATCC23877 were investigated. This research showed that the polymorphism in colony pigmentation had distinct features. The first aspect is the coexistence of four types of colonies: pigmented colonies (Pig+), pigment-defective colonies (Pigcol ), pigmented colonies harboring pigment-defective sectors (Pigsec+) or pigment defective papillae (Pigpap+). The second feature was revealed by the study on Pigpap+ colonies. We showed that WT progeny after 14 days of growth consisted almost totally of Pigpap+ colonies. Pigpap+ colonies were also found to be genetically different from each other. Characterization of twelve colonies presenting more than 20 papillae (Hyperpap colonies) led to the isolation of twelve mutator strains which produced at high frequency Pigcol- and Hyperpap colonies. Each exhibited a specific mutator phenotype and were distinct from each other. Such strains constituted a part of the polymorphism observed in each of the WT progeny and also generated a high variability. Finally, we showed that pigment-defective papillae were mutants and constituted a new form of genetic instability. PMID- 9748512 TI - Neuropeptide Y in relation to carbohydrate intake, corticosterone and dietary obesity. AB - Neuropeptide Y (NPY) is known to stimulate eating behavior and to be related to behavioral patterns of carbohydrate ingestion. The present report investigates this relationship further to: (1) characterize the specific NPY projection activated in different dietary paradigms; (2) understand associated changes in circulating hormones that may mediate dietary effects on NPY neurons; and (3) determine whether endogenous NPY in conditions with macronutrient diets can be linked to body fat. Male albino Sprague-Dawley rats were tested in two feeding paradigms, one in which the rats were given a choice of the macronutrients, carbohydrate, fat or protein, or the other involving a single diet varying in carbohydrate of fat content. These studies consistently demonstrated a close association between the ingestion of carbohydrate and NPY levels, specifically in the arcuate nucleus (ARC) and medial portion of the paraventricular nucleus (PVN) of the hypothalamus. In addition to revealing increased NPY activity in animals that naturally select high carbohydrate when given a choice of macronutrients, a single diet with 65% carbohydrate (10% fat), compared to a control diet with 45% carbohydrate (30% fat), significantly potentiates NPY gene expression and NPY immunoreactivity, as determined by in situ hybridization and immunohistochemistry. A further lowering of carbohydrate to 15% has little effect on NPY. Studies of medial hypothalamic fragments in vitro also reveal enhanced NPY release from hypothalamic tissue taken from rats maintained on high carbohydrate diet. Together with NPY, circulating corticosterone (CORT) levels are also highest in a high-carbohydrate condition and positively correlated with NPY in the ARC. An association between NPY and adiposity in these dietary conditions is indicated by significantly higher levels of NPY in the medial PVN in rats with high body fat, whether consuming a high-carbohydrate of high-fat diet. This evidence, linking NPY to carbohydrate intake and circulating CORT, suggests a role for this peptide in glucose homeostasis that is normally exhibited under conditions when carbohydrate stores are low. Disturbances in this homeostatic process, associated with hyperinsulinemia and higher levels of NPY, become evident with only a moderate rise in body fat on a high-carbohydrate as well as high-fat diet. PMID- 9748513 TI - Regulated expression during development and following sciatic nerve injury of mRNAs encoding the receptor tyrosine phosphatase HPTPzeta/RPTPbeta. AB - Three major isoforms of the receptor protein tyrosine phosphatase HPTPzeta/RPTPbeta (RPTPzeta/beta) have been previously identified, two with identical transmembrane and intracellular catalytic domains that differ by virtue of a long cysteine-free extracellular region, and a soluble proteoglycan called phosphacan that lacks the transmembrane and carboxy-terminal catalytic domains. To determine whether these RPTPzeta/beta variants are produced by alternative mRNA splicing of a common primary transcript, we performed genomic Southern analysis and characterized several rat cDNA and genomic RPTPzeta/beta clones. These studies indicated that the three major transcripts which encode phosphacan and the two RPTPzeta/beta phosphatase variants are encoded by a single gene, and further that additional alternative mRNA splicing is likely to result in the deletion of a 7 amino acid insert from the intracellular juxtamembrane region of both long and short phosphatase isoforms. Simultaneous quantitation of the three major isoforms by RNase protection analysis indicated that the mRNA encoding phosphacan had the highest relative abundance in the CNS while that encoding the short phosphatase isoform was most abundant relative to the other RPTPzeta/beta variants in the PNS. Following peripheral nerve crush, all RPTPzeta/beta mRNAs, including phosphacan and the phosphatase variants with and without the 21 base insert, were significantly induced in the distal segments of the sciatic nerve with a time course that correlated well with the response of Schwann cells to this injury. PMID- 9748514 TI - Identification of the creatine binding domain of creatine kinase by photoaffinity labeling. AB - A new photoaffinity probe with a benzophenone group, N-dibenzylphospho-N'-(4 benzoyl)-benzylguanidine (BzPG), has been synthesized on the basis on our previously described creatine kinase bisubstrate analog. BzPG is also a bisubstrate type analog whose photoinsertion is inhibited by the natural substrates of creatine kinase. When rabbit CK-MM is irradiated in the presence of BzPG then cleaved by CNBr, one labeled peptide can be purified by reverse phase HPLC and sequenced. This sequence of 31 amino acids (Ala30-Val60) contains a region which could be responsible for isoenzyme selectivity and another one just preceding the 11 amino acid peptide (Asp61-Thr70) very recently described as a putative creatine binding site. This second peptide was deduced from the comparison of 18 amino acid sequence alignments. We proposed the creatine binding site to be essentially a peptide from Lys39 to Val71. PMID- 9748515 TI - Mechanism of action of eukaryotic DNA topoisomerase I and drugs targeted to the enzyme. AB - DNA topoisomerase I is essential for cellular metabolism and survival. It is also the target of a novel class of anticancer drugs active against previously refractory solid tumors, the camptothecins. The present review describes the topoisomerase I catalytic mechanisms with particular emphasis on the cleavage complex that represents the enzyme's catalytic intermediate and the site of action for camptothecins. Roles of topoisomerase I in DNA replication, transcription and recombination are also reviewed. Because of the importance of topoisomerase I as a chemotherapeutic target, we review the mechanisms of action of camptothecins and the other topoisomerase I inhibitors identified to date. PMID- 9748516 TI - Correlations between DNA and cytogenetic damage induced after chemical treatment and radiation. AB - The induction of damage in human lymphocytes has been compared after treatment in vitro with two different agents, the chemical o-phenylenediamine (o-PDA) and gamma irradiation, in the alkaline single cell gel electrophoresis (Comet) assay, and after cytogenetic analysis. The chemical treatment caused dose-related increases in DNA damage in the Comet assay and cytogenetic damage in the first and second metaphases. The results revealed a very strong association between the two types of damage. Correlation coefficients were from 0.95 to 0.97. From previous studies, high correlation coefficients of 0.99 and 0.97 in the same assays were also evaluated for X-rays and fast neutrons, respectively. On the basis of such results, we suggest that the Comet assay responses provide a good prediction of cytogenetic damage. Thus, because of its simplicity and rapidity, the Comet assay would appear to be a very useful tool for determining the genotoxicity of environmental agents. PMID- 9748517 TI - Iron removal from monoferric human serum transferrins by 1, 2-dimethyl-3 hydroxypyridin-4-one, 1-hydroxypyridin-2-one and acetohydroxamic acid. AB - The kinetics of iron removal from both forms of human serum monoferric transferrin by three ligands, 1, 2-dimethyl-3-hydroxypyridin-4-one (L1), 1 hydroxypyridin-2-one and acetohydroxamic acid, have been evaluated at pH 7.4 and 25.0 degreesC. In almost all cases the rate of iron removal follows simple saturation kinetics with respect to the ligand concentration. No spectroscopically distinct intermediates are observed during the iron removal reaction, which is consistent with a mechanism in which the rate-limiting step in iron removal is a protein conformational change. In the presence of chloride or perchlorate, most systems continue to follow simple saturation kinetics, but with significantly different kmax values. Chloride accelerates iron release from both transferrin binding sites, while perchlorate accelerates iron release from the C terminal site but retards iron release from the N-terminal site. When the hydrochloride salt of L1 is used to prepare the L1 stock solution, the allosteric effect of the chloride produces a continuing increase in the rate of iron removal with increasing ligand concentration, so that one no longer observes simple saturation kinetics. A least squares fit of kobs vs. the ligand concentration for L1.HCl shows that the allosteric effect of the chloride not only enhances the first-order term for iron removal but also doubles the apparent kmax for the saturation term. This supports the view that allosteric binding of anionic ligands contributes to the observed variation in kmax among different ligands. A detailed description of this allosteric effect is not yet possible because the effect varies significantly from system to system, depending upon the specific anion that is binding at the allosteric site, the ligand that is used to remove the iron, and the transferrin lobe from which iron is removed. PMID- 9748518 TI - Barbiturates decrease the expression of vascular endothelial growth factor in hypoxic cultures of porcine brain derived microvascular endothelial cells. AB - Vascular endothelial growth factor (VEGF) is known to be produced in higher amounts during hypoxia by a variety of cell types and has been shown to increase the permeability of brain derived microvascular endothelial cells (BMEC) during hypoxia by an autocrine mechanism. Because the barbiturates, methohexital (MH) and thiopental (TP), induced a dose-dependent reduction in hypoxia-induced permeability changes of BMEC, the effect of both barbiturates on the VEGF expression during hypoxia was investigated. Both barbiturates decreased the hypoxia-induced expression of VEGF in BMEC in a concentration-dependent manner. This effect is partly caused by the impairment of the hypoxia-induced VEGF mRNA stabilization. VEGF-induced permeability changes during normoxia were unaffected by the barbiturates suggesting that MH and TP are directly reducing hypoxia induced VEGF synthesis. In conclusion, the inhibiting effect of these barbiturates on the hypoxia-induced VEGF expression results in the decreased permeability of the BMEC monolayer during hypoxia, which may contribute to the described neuroprotective action of barbiturates by reduction of brain edema formation. PMID- 9748519 TI - Fibroblast growth factor receptor-1 expression in the cortex and hippocampus in Alzheimer's disease. AB - Localization of fibroblast growth receptor (FGFR)-1 immunoreactivity was investigated immunochemically in postmortem brain tissue of Alzheimer's disease (AD) and age-matched control cases using a rabbit polyclonal antibody and a mouse monoclonal antibody specific for FGFR-1. In control cases, FGFR-1 immunoreactivity was identified in astrocytes in white matter and in hippocampal pyramidal neurons. In AD cases, the immunoreactivity in reactive astrocytes surrounding senile plaques was increased. The pattern of FGFR-1 immunoreactivity was confirmed in selected cases by in situ hybridization for FGFR-1 mRNA. Immunoreactivity using a monoclonal antibody demonstrated a similar distribution pattern. The localization of FGFR-1 is consistent with previous reports on the involvement of FGF-1 and FGF-2 in AD. PMID- 9748520 TI - Profiles of chemically-induced tumors in rodents: quantitative relationships. AB - The rodent carcinogenicity bioassay has been used for several decades for evaluating hundreds of chemicals, with the two aims of better understanding the etiologies of cancer, and of assessing the hazard posed by environmental and industrial chemicals. This has generated an enormous wealth of data and information on the phenomenon of chemical carcinogenicity. However, this information cannot be appreciated easily, since too many details may obscure the general trends present in the data; on the contrary, the use of computerized data analysis techniques suitable for the exploration of large databases makes its investigation much more fruitful, and its results more reliable. For this work, we collected a database of 536 rodent carcinogens, and we investigated the profiles of tumors (target organs) induced in the four experimental systems which are usually employed (rat and mouse, male and female). The analysis was performed with an Artificial Neural Network called Kohonen Self-Organizing Map, which is a computer-intensive method aimed at making the relevant information emerge automatically from the data itself. The analysis generated a global view, as well as a quantitative measure of the associations among the individual tumor types, and among the tumor profiles induced by the chemicals. In the complex interplay between the organ and species specificity of tumor induction, the species specificity generally overcame organ specificity, except for a few tumors (namely Lymphatic System, Brain, Forestomach, Stomach and Thyroid Gland). Moreover, the species specificity was remarkably stronger than the trans-species sex specificity. For three chemical classes (Aromatic Amines, Electrophilic/Alkylating Agents, Nitroarenes) most represented in the database, we investigated the hypothesis that a single mechanism of interaction with DNA would produce one, or a few very similar tumor profiles. Our analysis pointed out that no obvious association exists between chemical/mode of action class, and tumor profile. On the contrary, none of these classes induces a single tumor or pattern of tumors, but rather it appears that each class produces tumors at a wide range of sites. This suggests that an important determinant of the differences in tumor profile are the events that surround the ultimate mechanism of interaction with DNA. PMID- 9748521 TI - Marked, sustained expression of a novel 150-kDa oxygen-regulated stress protein, in severely ischemic mouse neurons. AB - The 150-kDa oxygen-regulated protein (ORP150) first was described with reference to the central nervous system in cultured astrocytes subjected to dense hypoxia. Subsequently its transcript was found in macrophages within human aortic atherosclerotic plaques, suggesting a role in protecting cells under hypoxic stress. In a mouse model of permanent focal brain ischemia, we aimed to elucidate the constitutive cellular localization in vivo of ORP150 in the central nervous system as well as the sequential alteration in its mRNA and protein expression during this severe ischemic insult. Immunohistochemical study demonstrated that ORP150 protein normally is present predominantly in neurons. The 78-kDa glucose regulated protein, which is another well-known stress protein retained in the endoplasmic reticulum, also was stained in neurons. During the first 3 h after ischemia, ORP150 antigenicity was markedly enhanced in severely damaged neurons, while the amount of the glucose-regulated protein was decreased. Preceding this change, orp150 mRNA was selectively induced in neurons undergoing postischemic cytoskeletal proteolysis, as early as 1 h after middle cerebral artery occlusion. These results indicated that ORP150 might be regulated by transcriptional level as for many stress proteins, but unlike previously described other stress proteins it was translated in the center of ischemic lesions despite nearly complete energy depletion. In this paper, the biological potentials of ORP150 protein in the setting of brain ischemia in vivo will also be discussed. PMID- 9748522 TI - Functional integration among 5-hydroxytryptamine receptor families in the control of female rat sexual behavior. AB - Serotonin (5-HT) receptor interaction in the control of female rat lordosis behavior was examined. Ovariectomized rats, with bilateral implants in the ventromedial nucleus of the hypothalamus (VMN), were hormonally primed with 25 micrograms estradiol benzoate and 500 micrograms progesterone. Rats were infused with the 5-HT3 receptor antagonist, 3-tropanyl-indole-3 carbonylate (tropisetron; 500 ng), or were coinfused with the 5-HT3 receptor antagonist and the 5-HT2A/2C receptor agonist, 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI; 500, 1500, or 2000 ng). Additional ovariectomized, hormone-primed rats received bilateral VMN infusions with the 5-HT1A receptor agonist, 8-hydroxy-2-(di-n-propylamino) tetralin (8-OH-DPAT; 200 ng), or were coinfused with the 5-HT1A receptor agonist and the 5-HT3 receptor agonist, m-chlorophenyl-biguanide (mCPBG; 250, 500, or 1000 ng). Lordosis behavior was observed prior to VMN infusion, during the infusion and for 30 consecutive minutes thereafter. Tropisetron reduced the lordosis to mount (L/M) ratio in every animal investigated but the decline was attenuated by coinfusion with DOI. Similarly, the L/M ratio declined following infusion with 8-OH-DPAT and the decline was dose-dependently reduced by coinfusion with mCPBG. Only the 5-HT3 receptor agonist altered the quality of the lordosis reflex. These studies provide evidence that the effects of 5-HT on female rat lordosis behavior involve the integrated activity of at least 3 different 5-HT receptor families. PMID- 9748523 TI - Crystal structure of bovine annexin VI in a calcium-bound state. AB - The crystal structure of a calcium-bound form of bovine annexin VI has been determined with X-ray diffraction data to 2.9 A by molecular replacement. Six Ca2+ ions were found, five in AB loops, one in a DE loop. Two loops (II-AB, which binds calcium, and V-AB, which does not) have conformations that differ significantly from those in calcium-free, human recombinant annexin VI. There are only small differences between the calci- and the apo-annexin VI in the rest of the molecule. Calcium by itself does not promote a major conformational change. PMID- 9748524 TI - Temporal characteristics of the protective effect of aminoguanidine on cerebral ischemic damage. AB - We investigated the temporal profile of the reduction in focal cerebral ischemic damage exerted by aminoguanidine (AG), an inhibitor of inducible nitric oxide synthase (iNOS). In anesthetized spontaneously hypertensive rats, the middle cerebral artery (MCA) was occluded distal to the origin of the lenticulostriate arteries. Rats were treated with vehicle (saline) or AG (100 mg kg-1, i.p.) immediately after MCA occlusion and, thereafter, two times per day. Rats were sacrificed 1(n = 7), 2(n = 8), 3 (n = 6) or 4 days (n = 5) after MCA occlusion. Injury volume (mm3) was determined in thionin-stained sections using an image analyzer. Volumes were corrected for ischemic swelling. Administration of AG up to 2 days after MCA occlusion did not reduce cerebral ischemic damage (p < 0.05 from vehicle; t-test). Treatment for a longer period decreased injury volume, the reduction averaging 21 +/- 5% at 3 days (p < 0.05) and 30 +/- 9% at 4 days (p < 0.05). Aminoguanidine did not affect ischemic brain swelling (p > 0.05). Administration of AG did not substantially modify arterial pressure, arterial blood gases, pH, hematocrit, plasma glucose and rectal temperature. We conclude that the protective effect of AG is time-dependent and occurs only when the drug is administered for longer than 2 days, starting after induction of ischemia. Because iNOS enzymatic activity develops more than 24 h after MCA occlusion [C. Iadecola, X. Xu, F. Zhang, E.E. El-Fakahany, M.E. Ross, Marked induction of calcium-independent nitric oxide synthase activity after focal cerebral ischemia, J. Cereb. Blood Flow, Metab. 14 (1995) 52-59; C. Iadecola, F. Zhang, X. Xu, R. Casey, M.E. Ross, Inducible nitric oxide synthase gene expression in brain following cerebral ischemia, J. Cereb. Blood Flow Metab. 15 (1995) 378-384.], the data support the hypothesis that the protective effect of AG is medicated by inhibition of iNOS in the post-ischemic brain. PMID- 9748525 TI - Clinical applications of the camptothecins. AB - The camptothecin topoisomerase I-targeting agents are new class of antitumor drugs with demonstrated clinical activity in human malignancies, such as colorectal cancer and ovarian cancer. Currently, irinotecan and topotecan are the most widely used camptothecin analogs in clinical use and clinical trials are ongoing to better characterize their spectra of clinical activity, to determine their optimal schedules of administration and to define their use in combination with other chemotherapeutic agents. Newer camptothecin analogs in clinical development, such as 9-aminocamptothecin, 9-nitrocamptothecin, GI147211 and DX 8951f, are also being studied to determine if they have improved toxicity and efficacy profiles compared with existing analogs. Other potential clinical applications include the use of camptothecin derivatives as radiation sensitizers or as antiviral agents. The successful development of the camptothecins as antitumor agents highlights the importance of topoisomerase I as a target for cancer chemotherapy. PMID- 9748526 TI - Phosphorylation and spatiotemporal distribution of KW8 (NDRF/NeuroD2), a NeuroD family basic helix-loop-helix protein. AB - KW8, a NeuroD family basic helix-loop-helix protein, was initially cloned during the course of screening for the genes related to long term potentiation in rat hippocampal slice. Its homologue NDRF/NeuroD was also reported. In this report its phosphorylation and spatiotemporal distribution was studied. KW8 was expressed not only during embryonic and neonatal periods but also in adults. In adult, KW8 was expressed only in brain tissues, such as the cerebral cortex, hippocampus and cerebellum. Immunohistological studies revealed that KW8 was localized in the nuclei of neurons. On immunoblotting of rat brain tissue, COS-1 cells and Neuro2A cells overexpressing KW8, this protein was detected as several diffuse bands. Alkaline phosphatase treatment reduced the molecular weights of these bands. Metabolic labeling with 32Pi in COS-1 cells confirmed that the KW8 protein was phosphorylated in vivo. Some of the physiological functions of KW8 might be regulated by this phosphorylation. In yeast, the GAL4 fusion protein containing the C-terminal region of KW8 activated transcription of the reporter gene, suggesting that KW8 had transcriptional activity. PMID- 9748527 TI - Enhanced sensitivity to topoisomerase inhibitors in synchronous CHO cells pre treated with 5-azacytidine. AB - Multidrug combination has been shown to be very useful to improve antitumor activity as well as to reduce the toxicity of different anti-cancer drugs. We have evaluated the interaction between the hypomethylating agent 5-azacytidine and the topoisomerase I and topoisomerase II inhibitors Camptothecin (CPT) and 4' (9-acridinylamino) methanesulfon-m-anisidide (m-AMSA) respectively, based on the hypothesis that through the alteration of chromosome replication timing following DNA hypomethylation, the number of replication forks in early S phase might increase, so enhancing the probability of a collision between a blocked cleavable complex (DNA-topo I-CPT or DNA-topo II-m-AMSA) and a replication fork. We have tested the capacity of CPT and m-AMSA to induce chromosomal aberrations as well as reproductive cell death in synchronous cultured Chinese hamster ovary cells after a pretreatment with 5-azacytidine with positive results. PMID- 9748528 TI - Systemic d-amphetamine administration causes a reduction of kynurenic acid levels in rat brain. AB - Tissue levels of the endogenous excitatory amino acid receptor antagonist kynurenic acid (KYNA) and of its bioprecursor L-kynurenine were measured in rats of different ages after d-amphetamine administration. In adult animals, extracellular KYNA concentrations were also determined in vivo by hippocampal microdialysis. In the adult brain, d-amphetamine caused a transient, dose dependent decrease in tissue content and extracellular levels of KYNA, reaching a nadir of approximately 70% of control values after 1 h at 5 mg/kg. Quantitatively similar decrements were observed in four different brain regions. Seven, 14 and 28-day-old pups were particularly sensitive to the drug, showing a reduction in forebrain KYNA levels to 25%, 40% and 35% of control values, respectively, 1 h after the administration of 5 mg/kg d-amphetamine. Notably, no changes in brain L kynurenine levels and in liver L-kynurenine and KYNA concentrations were found after d-amphetamine administration. Thus, endogenous monoamines released by d amphetamine may interfere with the transamination of L-kynurenine to KYNA specifically in the brain. These results suggest that d-amphetamine increases excitatory amino acid receptor function temporarily by reducing the levels of endogenous KYNA. PMID- 9748529 TI - Activation of gadd153 expression through transient cerebral ischemia: evidence that ischemia causes endoplasmic reticulum dysfunction. AB - The expression of the gene encoding the C/EBP-homologous protein (CHOP), which is also known as growth arrest and DNA-damage-inducible gene 153 (gadd153), has been shown to be specifically activated under conditions that disturb the functioning of the endoplasmic reticulum (ER). To investigate a possible role of ER dysfunction in the pathological process of ischemic cell damage, we studied ischemia-induced changes in gadd153 expression using quantitative PCR. Transient cerebral ischemia was produced in rats by four-vessel occlusion. In the hippocampus, ischemia induced a pronounced increase in gadd153 mRNA levels, peaking at 8 h of recovery (6.4-fold increase, p<0.01), whereas changes in the cortex were less marked (non-significant increase). To elucidate the possible mechanism underlying this activation process, gadd153 mRNA levels were also evaluated in primary neuronal cell cultures under two different conditions, both leading to a depletion of ER calcium pools in the presence or absence of an increase in cytoplasmic calcium activity. The first procedure, exposure to thapsigargin, an irreversible inhibitor of ER Ca2+-ATPase, caused a marked increase in gadd153 mRNA levels both in cortical and hippocampal neurons, peaking at 12-18 h after treatment. The second procedure, immersion of cells in calcium free medium supplemented with EGTA, caused only a transient increase in gadd153 mRNA levels, peaking at 6 h of recovery, indicating that a depletion of ER calcium stores in the absence of an increase in cytoplasmic calcium activity is sufficient to activate neuronal gadd153 expression. The results imply that transient cerebral ischemia disturbs the functioning of the ER and that these pathological changes are more pronounced in the hippocampus compared to the cortex. PMID- 9748530 TI - Positive responses to carcinogens in the yeast DEL recombination assay are not due to selection of preexisting spontaneous revertants. PMID- 9748531 TI - Identification of rat brain p42(IP4), a high-affinity inositol(1,3,4, 5)tetrakisphosphate/phosphatidylinositol(3,4,5)trisphosphate binding protein. AB - Inositol(1,3,4,5)tetrakisphosphate (InsP4) and phosphatidylinositol(3,4,5)trisphosphate (PtdInsP3) are two potential second messengers with a still largely unknown mode of action. We recently cloned the 42 kDa protein p42IP4 previously purified from pig cerebellum, which binds InsP4 (Kd approximately 2 nM) and PtdInsP3 with comparable affinities (Stricker et al., FEBS Lett. 405 (1997) 229). The protein p42IP4 (pig) is highly homologous to centaurin-alpha, a larger protein of 46 kDa, derived from a rat brain cDNA library clone (Hammonds-Odie et al., J. Biol. Chem. 271 (1996) 18859). Here we investigated whether also p42IP4 is expressed in rat brain and how it might be related to centaurin-alpha. When we carried out RT-PCR using mRNA from brain of rats of different ages we obtained several clones corresponding to p42IP4, but not to centaurin-alpha. The existence of p42IP4 in rat brain is supported by the following findings: (1) biochemical analysis of the purified rat brain protein shows inositol phosphate ligand affinities identical to those of the protein from other species; (2) Western blot analysis of rat brain membrane fractions using a peptide-specific antiserum revealed only the 42 kDa protein (p42IP4), but did not give evidence for the occurrence of a larger 46 kDa centaurin-alpha-like protein in rat brain; and (3) the amino acid sequences deduced from p42IP4 cDNA are highly homologous in several species and are confirmed by protein fragment microsequences. Thus, p42IP4 from rat brain which has two pleckstrin homology domains is a protein largely conserved between different species and most likely has an important function in inositol phosphate or inositol lipid signal transduction. PMID- 9748532 TI - Dose-response relationships in chemical carcinogenesis: superposition of different mechanisms of action, resulting in linear-nonlinear curves, practical thresholds, J-shapes. AB - The shape of a carcinogen dose-cancer incidence curve is discussed as the result of a superposition of dose-response relationships for various effects of the carcinogen on the process of carcinogenesis. Effects include direct DNA damage, e.g., by covalent binding, indirect DNA damage, e.g., by increased formation of reactive oxygen species or interaction with DNA replication or chromosome integrity. The 'fixation' of a DNA adduct as a heritable mutation depends on its pro-mutagenic potency and on the rates of DNA repair and DNA replication. Endogenous and unavoidable DNA damage is responsible for a background rate of the process of mutagenesis and carcinogenesis and forms the basis of spontaneous cancer incidence. For DNA-reactive carcinogens, linearity of the dose response at the low-dose end is expected. With increasing dose, saturation of DNA repair can introduce a sublinearity (example: dimethylnitrosamine). Stimulation of cell division as a result of high-dose toxicity and regenerative proliferation also results in a sublinear deviation from low-dose linearity. If the DNA-damaging potency of the carcinogen is low in comparison with the high-dose effects, the linear part of the low dose-cancer incidence curve might be hidden within the background variability. Under such conditions, 'practical thresholds' could be discussed (formaldehyde). If a carcinogen increases the rate of cell division or the level of oxidative stress at high dose but has an antimitogenic or antioxidative effect at low dose, a J-shaped (or: U-shaped) curve with a decrease of the spontaneous tumor incidence at low dose could result (caffeic acid; TCDD). This phenomenon has been observed even under conditions of a genotoxic contribution (ionizing radiation; diesel exhaust particles). For a mechanism based assessment of a low-dose cancer risk, information on the various modes of action and modulations should be available over the full dose range, and models should be refined to incorporate the respective information. PMID- 9748533 TI - Pick's disease: cytoskeletal changes in the hypothalamic lateral tuberal nucleus. AB - Basolateral portions of the human hypothalamus contain an extended nuclear gray, the lateral tuberal nucleus (LTN), which undergoes conspicuous pathological changes in a number of neurodegenerative diseases. The present study points to the severe affliction of this nucleus in Pick's disease (PID). Immunoreactions for abnormally phosphorylated tau-protein permit identification of the permutations. Only a fraction of the abnormal fibrillary material developing in the course of the disease shows a pronounced argyrophilia. Key features are the Pick bodies (PBs) which contain an argyrophilic material. Unusual non-spherical PBs develop in the LTN as flat structures with peripheral indentations. Small teardrop-like Pick neurites (PNs) emerge in varicose widenings of neuronal processes and display a much weaker argyrophilia. The characteristic alterations seen in PID reliably can be differentiated from lesions of the LTN which slowly emerge in the course of Alzheimer's disease (AD). PMID- 9748534 TI - A comparative study of in vivo mutation assays: analysis of hprt, lacI, cII/cI and as mutational targets for N-nitroso-N-methylurea and benzo[a]pyrene in Big Blue mice. AB - We have compared the response of the native hprt gene and the lacI, cII, and cI transgenes in Big Blue B6C3F1 mice following treatment with either N-nitroso-N methylurea (MNU) or benzo[a]pyrene (BaP). Three weeks after mutagen treatment splenic T cells were isolated from the animals, and samples were either cultured to measure mutation at the native hprt locus or used to extract genomic DNA for transgene mutation analysis. Phage rescued from extracted DNA were plated in the presence of 5-bromo-4-chloro-3-indolyl-beta-d-galactopyranoside (X-gal) to score lacI mutations, or plated on a hflAB lawn to score cII and cI mutants. With MNU hprt mutant frequency increased in a dose-related, sublinear manner up to 78-fold above background at the highest dose tested (20 mg/kg). In comparison, the lacI transgene yielded only a 3.1-fold increase at this dose, and the cII and cI transgenes did not show any increase. With 150 mg/kg BaP a 5.8- and 8.7-fold increase in mutant frequency was observed at hprt and lacI, respectively, while only a 1.3-fold increase was observed at cII. DNA sequencing revealed an increase in GC-->TA transversions among the cII mutants, suggesting that the increase was related to BaP exposure. No significant increase in cI mutant frequency was observed. Therefore, the order of mutation assay sensitivity was hprt>lacI>cII/cI with MNU, and hprt approximately lacI> cII/cI with BaP. While the hflAB selection system offers significant advantages with respect to cost and effort when compared to the lacI assay, additional evaluation of its sensitivity is warranted. PMID- 9748535 TI - Physiological regulation of eukaryotic topoisomerase II. AB - Topoisomerase II is an essential enzyme in all organisms with several independent roles in DNA metabolism. In this article we review our knowledge on the regulation of the expression and catalytic activity of topoisomerase II in both lower and higher eukaryotes. Current data indicate that the regulation of topoisomerase II gene expression is complex, with positive and negative controls in evidence at the level of both promoter activity and mRNA stability. Similarly, the activity of the mature enzyme can be regulated by the action of several different protein kinases. Of particular interest is the cell cycle-dependent phosphorylation of topoisomerase II, including multiple, mitosis-specific modifications, which are proposed to regulate the essential chromosome decatenation activity of the enzyme. PMID- 9748536 TI - Early localization of mRNA coding for 5-HT1A receptors in human brain during development. AB - The distribution of 5-HT1A receptor mRNA in the human brain was studied in neonatal, children and adult cases by means of in situ hybridization histochemistry, using an oligonucleotide derived from the coding region of the human receptor. A prenatal pattern of development was observed. The hippocampus, raphe nuclei and neocortex presented high levels of hybridization already at the fetal/neonatal stage, fully comparable to the adult. A high and transient hybridization signal was found in cerebellum. These results support a role for 5 HT1A receptors in the regulation of neural development. PMID- 9748538 TI - Pregnenolone reverses the age-dependent accumulation of glial fibrillary acidic protein within astrocytes of specific regions of the rat brain. AB - Although aged-related modifications of astrocytes have been frequently described, little is known so far about the signals responsible for these modifications. Since it is well demonstrated that astrocytes are highly responsive to a variety of steroids, we hypothesized that modifications of cerebral astrocytes may result from the age-related decrease of circulating steroids. In the present study, we investigated the effects of the chronic administration of pregnenolone (PREG), the precursor of all steroid hormones, on the age-related extension of astrocytic processes in various brain regions. In adult (2-3 month-old) and aged (22-24 month-old) rats, quantitative image analysis was used to estimate, within each region, the number of astrocyte cell bodies immunostained (IS) for S100, and the surface occupied by astrocytic cell bodies and processes IS for glial fibrillary acidic protein (GFAP). In all regions, the surface occupied by GFAP-IS structures was increased in the aged vs. the adult rats, whereas no significant modifications were observed in the number of S100-IS cell bodies. Chronic administration of PREG to aged rats induced a marked decrease in the surface occupied by GFAP-IS structures in the cortex, amygdala and thalamus, without any significant effect on the number of S100-IS cell bodies present in these regions. By contrast, PREG had no significant effect when administered to adult animals. These data suggest that decreased levels of circulating steroid hormones may be responsible for the age-dependent modifications of the astrocytes present in various brain regions, and that these modifications can be at least partly corrected by the administration of PREG. PMID- 9748537 TI - Comparison of DNA adduct levels associated with oxidative stress in human pancreas. AB - DNA adducts associated with oxidative stress are believed to involve the formation of endogenous reactive species generated by oxidative damage and lipid peroxidation. Although these adducts have been reported in several human tissues by different laboratories, a comparison of the levels of these adducts in the same tissue samples has not been carried out. In this study, we isolated DNA from the pancreas of 15 smokers and 15 non-smokers, and measured the levels of 1,N6 etheno(2'-deoxy)guanosine (edA), 3, N4-etheno(2'-deoxy)cytidine (edC), 8-oxo-2' deoxyguanosine (8-oxo-dG), and pyrimido[1,2-alpha]purin-10(3H)-one (m1G). Using the same DNA, the glutathione S-transferase (GST) M1, GSTT1, and NAD(P)H quinone reductase-1 (NQO1) genotypes were determined in order to assess the role of their gene products in modulating adduct levels through their involvement in detoxification of lipid peroxidation products and redox cycling, respectively. The highest adduct levels observed were for m1G, followed by 8-oxo-dG, edA, and edC, but there were no differences in adduct levels between smokers and non smokers and no correlation with the age, sex or body mass index of the subject. Moreover, there was no correlation in adduct levels between edA and eC, or between edA or edC and m1G or 8-oxo-dG. However, there was a significant correlation (r=0.76; p<0.01) between the levels of 8-oxo-dG and m1G in human pancreas DNA. Neither GSTM1 nor NQO1 genotypes were associated with differences in any of the adduct levels. Although the sample set was limited, the data suggest that endogenous DNA adduct formation in human pancreas is not clearly derived from cigarette smoking or from (NQO1)-mediated redox cycling. Further, it appears that neither GSTM1 nor GSTT1 appreciably protects against endogenous adduct formation. Together with the lack of correlation between m1G and edA or edC, these data indicate that the malondialdehyde derived from lipid peroxidation may not contribute significantly to m1G adduct formation. On the other hand, the apparent correlation between m1G and 8-oxo-dG and their comparable high levels are consistent with the hypothesis that m1G is formed primarily by reaction of DNA with a base propenal, which, like 8-oxo-dG, is thought to be derived from hydroxyl radical attack on the DNA. PMID- 9748539 TI - Conditional coupling of striatal dopamine D1 receptor to transcription factors: ontogenic and regional differences in CREB activation. AB - The coupling of striatal dopamine D1 receptors to c-fos transcription exhibit all or-none regional and ontogenic differences: the D1 agonist SKF 38393 fails to induce c-fos expression in the striatum, except during the early postnatal period in the striosomes, or in the caudal extremity of the striatum in adult animals. In an attempt to better delineate the mechanism responsible for interrupting or enabling this conditional coupling of D1 receptors to c-fos transcription we have examined, through immunocytochemistry and gel shift assay, the activation of the cyclic AMP-response element binding protein (CREB) transcription factor in response to the D1 agonist in the murine striatum. Phosphorylated-CREB (P-CREB) immunoreactivity in response to the dopamine D1 agonist (+/-)SKF 38393 (15 mg/kg, i.p.) was prominent in the caudal extremity of the striatum in adult animals (P90). In neonatal (P5) mice, P-CREB immunoreactive neurons were observed both in the caudal and in the rostral parts of the striatum, without obvious patchy distribution. Gel shift assays performed on nuclear protein extracts from either the rostral or the caudal part of striatal tissue of neonatal (P5) or adult (P90) mice provided quantitative assessment, showing differences both in the amplitude and in the time course of the response, since P-CREB binding in adults culminated 45 min after (+/-)SKF 38393 (15 mg/kg, i.p.) injection, wheareas the peak value appeared as soon as 10 min after injection in P5 mouse pups, suggesting the involvement of partly distinct transduction pathways. PMID- 9748540 TI - 1H-NMR on line monitoring of water activity during lipase catalysed esterification. AB - 1H-NMR spectroscopy is used to determine simultaneously the water activity (aw) and the time course of an esterification reaction catalysed by a lipase. Chemical shifts signals of hydroxylic hydrogens in fast exchange (i.e the average hydroxylic signal of acid, alcohol and water) varies with water activity and ester content. Calibration curves have been established from model media composed of the substrates and various ester contents, at different water activities, in order to mimic a reaction medium. One relationship is established between water activity, hydroxylic hydrogen signal chemical shift and ester content. In order to estimate the water activity evolution as a function of time, this last relationship is applied to the hydroxylic hydrogen chemical shift measured in a reaction medium where the Rhizomucor miehei lipase in a powder form is suspended in the liquid substrates. This alternative way of determining the water activity based on hydroxylic hydrogen chemical shift presents some advantages over more classical means, i.e. time saved and inaccuracies avoided by monitoring without handling the sample. PMID- 9748541 TI - Sex differences in estrogenic regulation of preproenkephalin mRNA levels in the medial preoptic area of prepubertal rats. AB - Opioids have been implicated in sexual differentiation of the brain and in the regulation of reproductive behavior and endocrinology of mammals. Previous studies have indicated that estrogen administration in adults regulates preproenkephalin MRNA levels in several hypothalamic brain nuclei. We have determined preproenkephalin mRNA levels in estrogen-treated juvenile male and female rats to investigate the developmental pattern of estrogenic regulation of enkephalinergic neurons in the medial preoptic area. Rats were treated with estradiol benzoate (20 microgram/kg/day) or oil from day 21 to 23. Sections of the medial preoptic area (mPOA) were studied by in situ hybridization histochemistry at the single cell level and quantified with the assistance of an image analysis system. Our data indicate that males contain higher levels of preproenkephalin mRNA per neuron than females. In addition, our results indicate that estrogen causes an upward shift in the amount of mRNA expressed per cell, females demonstrating a greater response to estrogen than males. An increase in soma cell area following estrogen treatment was observed only in female mPOA enkephalinergic neurons. Taken together, these results indicate a sex difference in total preproenkephalin levels and in estrogenic regulation of preproenkephalin mRNA in the POA of juvenile rats. These results are discussed in relation to the differential role opioids may play in male and female reproductive physiology. PMID- 9748542 TI - Propranolol attenuates haloperidol-induced Fos expression in discrete regions of rat brain: possible brain regions responsible for akathisia. AB - Neuroleptics induce several extra-pyramidal side effects, such as akathisia, acute dystonia and parkinsonism. Although recently developed atypical neuroleptics ameliorate some of these side effects, akathisia remains a common and severely distressing adverse reaction. Several drugs are reported to be of clinical use for the pharmacological treatment of akathisia. In particular, the beta-adrenoceptor blocker, propranolol, has been widely used for the treatment of akathisia, but it does not ameliorate other extra-pyramidal side effects. To identify the neural substrates of akathisia, we investigated the effects of propranolol on haloperidol-induced Fos expression in rat brain. Haloperidol (1 mg/kg) induced Fos-positive nuclei in several regions of the brain, including the cingulate cortex area 3, piriform cortex nucleus accumbens, caudate-putamen, ventral lateral septum and parietal cortex. Pretreatment with propranolol (5 mg/kg) reduced the number of Fos-positive nuclei in the cingulate cortex area 3, the piriform cortex and area 1 of the parietal cortex. Injection of vehicle by itself tended to increase Fos expression in the cingulate cortex area 3 and the piriform cortex. Considering the functions of these brain regions, we speculate that the most plausible neural framework for haloperidol-induced akathisia involves area I of the parietal cortex, but possible roles for the cingulate cortex area 3 and the piriform cortex cannot be ruled out. PMID- 9748543 TI - DNA adducts in humans after exposure to methylating agents. AB - Human exposure to methylating agents appears to be widespread, as indicated by the frequent occurrence of methylated DNA adducts in human DNA. The high incidence of methylated DNA adducts even in humans thought not to have suffered extensive exposure to environmental methylating agents implies that chemicals of endogenous origin, probably N-nitroso compounds such as the strongly carcinogenic N-nitrosodimethylamine (NDMA), may be primarily responsible for their formation and raises the question of the carcinogenic risks associated with such exposure. In addition to accumulation of DNA damage, other factors (such as induced cell proliferation) appear to be important in determining the probability of induction of mutation or cancer by NDMA, implying that high to low dose risk extrapolations should not be based on the assumption of dose- or even adduct-linearity. Comparative studies of the accumulation and repair of methylated adducts in humans and animals treated with methylating cytostatic drugs do not reveal significant species differences. Based on this and the dosimetry of adduct accumulation in rats chronically exposed to very low doses of NDMA, it is suggested that the exposure needed to account for the levels of adducts found in human DNA may be of the order of hundreds of micrograms NDMA (or equivalent) per day, a level of exposure which may well represent a significant carcinogenic hazard for man. PMID- 9748544 TI - The role of tryptophan residues in Escherichia coli arginyl-tRNA synthetase. AB - The effect of N-bromosuccinimide (NBS) on the activity of Escherichia coli arginyl-tRNA synthetase (ArgRS) was studied. The results showed that only one tryptophan residue was easy of access to the reagent and was closely related to enzyme activity. When all the five tryptophan residues in ArgRS were changed via site-directed mutagenesis singly into Ala, the aminoacylation activity of the Trp162 mutated enzyme decreased seriously, while the other four mutant enzymes retained almost the same activity as the native one. The oxidation of the five mutant enzymes with NBS demonstrated that only the mutation of Trp162 resulted in the loss of sensitivity to the reagent. These results strongly suggest that Trp162 is more accessible to NBS and is related to enzyme activity. Furthermore, the far-UV CD spectroscopy of the mutant enzyme ArgRS162WA showed little change in its secondary structure. Finally, studies on the kinetics of the mutant enzyme ArgRS162WA in aminoacylation reaction showed that the reduction in activity could be attributed to the decrease in the values of kcat and kcat/Km for arginine. The thermodynamic calculation indicates that this mutation causes a decrease of the binding energy by 2.7 kJ/mol. Our data suggest that Trp162 is involved in the binding of arginine and in the transition state stabilization. PMID- 9748545 TI - Mechanism of action of eukaryotic topoisomerase II and drugs targeted to the enzyme. AB - Topoisomerase II is a ubiquitous enzyme that is essential for the survival of all eukaryotic organisms and plays critical roles in virtually every aspect of DNA metabolism. The enzyme unknots and untangles DNA by passing an intact helix through a transient double-stranded break that it generates in a separate helix. Beyond its physiological functions, topoisomerase II is the target for some of the most active and widely prescribed anticancer drugs currently utilized for the treatment of human cancers. These drugs act in an insidious fashion and kill cells by increasing levels of covalent topoisomerase II-cleaved DNA complexes that are normally fleeting intermediates in the catalytic cycle of the enzyme. Over the past several years, we have made considerable strides in our understanding of the catalytic mechanism of topoisomerase II and the mechanism of action of drugs targeted to this enzyme. These advances have provided novel insights into the physiological functions of topoisomerase II and have led to the development of more efficacious chemotherapeutic regimens and novel anticancer drugs. Considering the importance of topoisomerase II to the eukaryotic cell and to cancer chemotherapy, it is essential to understand its enzymatic function and pharmacological properties. Therefore, this review will discuss the mechanism of action of eukaryotic topoisomerase II and topoisomerase II-targeted drugs. PMID- 9748546 TI - Ovarian steroids differentially regulate the expression of PRL-R in neuroendocrine dopaminergic neuron populations: a double label confocal microscopic study. AB - The aims of this study were (1) to identify the possible hypothalamic targets for a short prolactin (PRL) feedback in the adult female rat by identifying DAergic neuron populations expressing PRL receptor (PRL-R); (2) to describe the effect of ovarian steroids on the expression of PRL-R and (3) to compare the distribution of both the extracellular (EC) and ligand binding (LB) domains of the PRL-R on the hypothalamic dopaminergic neurons by applying double label immunocytochemistry for the different domains of PRL-R and for tyrosine hydroxylase (TH). Five- to six-month-old female rats were ovariectomized (OVX) and implanted with either 17 beta-estradiol (E2), progesterone (P4) or received an E2 and a P4 implant (E2 + P4) at the same time. In the periventricular nucleus and in the dorsomedial portion of the middle arcuate nucleus, a dramatic increase in PRL-REC immunoreactivity was observed in E2 implanted rats. This increase was attenuated in E2 + P4 rats, but P4 treatment alone had no effect. Changes in PRL REC expression were paralleled by changes in serum PRL levels. Interestingly, PRL REC expression in the rostral arcuate nucleus decreased in P4 implanted rats, however, P4 did not attenuate the E2-induced increase in PRL-REC density. PRL-REC immunostaining was observed on the membrane, in the cytoplasm and in the nucleus. PRL-RLB immunoreactivity was also detectable in the TH positive neurons, but no nuclear staining was observed with this antibody. However, we found a strong PRL RLB immunostaining in the ependymal lining of the 3rd ventricle and in the processes of tanycytes projecting to the median eminence. These data indicate that (1) all neuroendocrine DAergic cells can be targets for PRL, (2) expression of PRL-R is differentially affected by ovarian steroids in the different TH cell populations, (3) PRL-RLB domain may be involved in trafficking PRL in the median eminence. PMID- 9748547 TI - Molecular cloning in the marmoset shows that semenogelin is not the precursor of the TRH-like peptide pGlu-Glu-Pro amide. AB - Two peptides with similar structures to thyrotropin-releasing hormone (TRH), pGlu Glu-Pro amide and pGlu-Phe-Pro amide, have been identified in human seminal fluid and it has been shown that one of these peptides, pGlu-Glu-Pro amide, has the ability to increase the capacitation of sperm cells, consistent with a role in fertility. In order to select a species in which there is a high degree of expression of the genes that code for 'TRH-like' peptides, we have determined the levels of these peptides in the prostate, pancreas and thyroid of a range of species including rat, rabbit, ox, marmoset, macaque and man. The peptides were extracted from the tissues and purified before determination by RIA with TRH antibody. In addition, trypsin digestion and TRH RIA was used to investigate the presence of N-extended forms. The highest concentrations of TRH-immunoreactive peptides were found in the tissues of the marmoset, Callithrix jacchus. Ion exchange chromatography demonstrated that marmoset thyroid contained principally authentic TRH, the pancreas contained both TRH and TRH-like peptides while the prostate contained TRH-like peptides alone. Further purification by HPLC showed that the main TRH-immunoreactive peptide in marmoset prostate was pGlu-Glu-Pro amide and a second component was identified as pGlu-Phe-Pro amide. The results indicate that the biosynthesis of these peptides could be studied to advantage in the marmoset. The biosynthetic precursors of the TRH-like peptides have not been identified. To examine whether pGlu-Glu-Pro amide might originate from semenogelin, we determined the sequence of semenogelin in the marmoset. It exhibited a high degree of homology with human semenogelin-I, but in place of the Lys-Gln-Glu-Pro sequence that might give rise to pGlu-Glu-Pro amide, marmoset semenogelin possessed the sequence Ser-Gln-Asp-Gln which cannot serve as a precursor for a TRH-like peptide. Further evidence was obtained by Northern blot analysis of a range of marmoset tissues. The results showed that semenogelin is not present in marmoset prostate. It is concluded that pGlu-Glu-Pro amide originates from a precursor distinct from semenogelin, both in marmoset and in man. PMID- 9748548 TI - On the origins of tumor mutations in cancer genes: insights from the p53 gene. PMID- 9748549 TI - Characterization of the hatching enzyme from embryos of an anuran amphibian, Rana pirica. AB - The culture medium in which prehatching embryos of the frog, Rana pirica, were cultured (hatching medium) solubilized the vitelline coat (VC) of unfertilized eggs and contained molecules reactive to antibodies (anti-UVS.2) against the Xenopus hatching enzyme (HE). The hydrolyzing activity of the hatching medium against Pro-Phe-Arg-MCA was inhibited dose-dependently by the same antibodies. Using anti-UVS.2 as a probe, we purified two distinct 56 kDa molecules exhibiting Pro-Phe-Arg-MCA hydrolyzing activity. These 56 kDa molecules, which were separable on anion exchange chromatography, were the same with respect to VC solubilizing activity and a substrate specificity for various MCA-peptides, and they were regarded as charge isomers that function as the HE. The hydrolyzing activity against Pro-Phe-Arg-MCA of HE was optimal at pH of 7.6, with the apparent Km value of 250 microM at 30 degreesC. The activity was strongly inhibited by DFP and EDTA, and was accelerated by extremely low concentrations of Mg2+ and Zn2+, indicating the serine protease and metalloprotease nature of the HE. The HE was glycosylated and was present as a putative proenzyme form of 63 kDa. PMID- 9748550 TI - Protein kinase C: a paradigm for regulation of protein function by two membrane targeting modules. PMID- 9748551 TI - Chronic low-dose lesion equilibrium along genes: measurement, molecular epidemiology, and theory of the minimal relevant dose. AB - Spontaneous mutations seem to be caused almost entirely by endogenous lesions. The pattern of these lesions along a gene represents an equilibrium between damage and repair. A pattern can be measured using ligation-mediated PCR (polymerase chain reaction) and a large chronic dose of a suspected endogenous mutagen. A study using dimethylsulfate-induced 7meGuanine lesions indicates that the exogenously induced pattern depends on how methyl purine glycosylase recognizes sequence context and, for this lesion, the pattern may be independent of the mutagen's dose. PMID- 9748552 TI - Catalytic inhibitors of DNA topoisomerase II. AB - Catalytic inhibitors of mammalian DNA topoisomerase II have been found recently in natural and synthetic compounds. These compounds target the enzyme within the cell and inhibit various genetic processes involving the enzyme, such as DNA replication and chromosome dynamics, and thus proved to be good probes for the functional analyses of the enzyme in a variety of eukaryotes from yeast to mammals. Catalytic inhibitors were shown to be antagonists against topoisomerase II poisons. Thus bis(2,6-dioxopiperazines) have a potential to overcome cardiac toxicity caused by potent antitumor anthracycline antibiotics such as doxorubicin and daunorubicin. ICRF-187, a (+)-enantiomer of racemic ICRF-159, has been used in clinics in European countries as cardioprotector. Furthermore, bis(2,6 dioxopiperazines) enhance the efficacy of topoisomerase II poisons by reducing their side effects in preclinical and clinical settings. Bis(2,6 dioxopiperazines) per se among others have antitumor activity, and one of their derivatives, MST-16 or Sobuzoxane, bis(N1-isobutyloxycarbonyloxymethyl-2, 6 dioxopiperazine), has been developed in Japan as an anticancer drug used for malignant lymphomas and adult T-cell leukemia in clinics. PMID- 9748553 TI - Studies on the interaction between 1,2,3,4-tetrahydro-beta-carboline and cigarette smoke: a potential mechanism of neuroprotection for Parkinson's disease. AB - 1,2,3,4-Tetrahydro-beta-carboline (TH beta C) is an endogenous or environmental neurotoxic factor putatively involved in the development of Parkinson's disease (PD). As part of our efforts to characterize the mechanism of the reported protection of smoking against PD, we have examined the interaction between TH beta C and cigarette smoke. We found that TH beta C reacts in vitro and under physiological conditions with some components of cigarette smoke to form N2 (cyanomethyl)-TH beta C (CM-TH beta C), N2-(gamma-cyanoethyl)-TH beta C (CE-TH beta C), N2-(1'-cyanopropyl)-TH beta C (CP-TH beta C), N2-(1'-cyanobutyl)-TH beta C (CB-TH beta C) and N2-formyl-TH beta C (F-TH beta C). Significant differences in the recovery of some of these TH beta C-derivatives were obtained for Burley and Bright tobacco. Several of the reported compounds showed reversible and competitive MAO-A inhibitory properties. The detection of some of these compounds in rat brain after chronic administration of TH beta C and a solution of cigarette smoke proved that the reported interactions also occur in vivo. These results are discussed as a potential mechanism of neuroprotection in the development of PD. PMID- 9748554 TI - Radiation induced chromosomal instability in human T-lymphocytes. AB - Chromosomal instability in proliferating mammalian cells is characterized by a persistent increase of chromosomal aberrations and rearrangements occurring de novo during successive cell generations. Recent results from many laboratories using a variety of cells and cytogenetic end points show that this phenotype can be induced by low as well as high LET irradiation. A typical feature of chromosomal instability in primary human G0-lymphocytes exposed to gamma irradiation at both high dose rate (45 Gy h-1) and low dose rate (0.024 Gy h-1) is the appearance of novel aberrations in the clonal progeny of the irradiated cell, many generations after the exposure. The same phenotype was observed in lymphocytes that were allowed to recover for 5 days in G0 after the radiation exposure, as well as in hprt-mutant T cell clones. These results demonstrate that neither the acute genotoxic stress caused by high dose rate as compared to low dose rate irradiation, nor a hypothesized conflict between mitogen induced growth stimulation and growth arrest due to radiation damage, seem to be critical conditions for the development chromosomal instability in these cells. In contrast to observations in other cells, no evidence of a persistent decrease of cloning ability was observed in the progeny of radiation-exposed human lymphocytes, and no alteration was observed in their sensitivity to a second radiation exposure. Furthermore, the frequency of CA-repeat length variation at three loci was not increased in the progeny of X-irradiated T cells as compared to non-irradiated cells, which indicates that microsatellite instability is not part of the chromosomal instability phenotype in human T-lymphocytes. PMID- 9748555 TI - Characterization of neurons in the area of the medullary lateral reticular nucleus responsive to noxious visceral and cutaneous stimuli. AB - In halothane-anesthetized rats, 283 caudal medullary neurons responsive to colorectal distension (CRD) were characterized using extracellular electrodes. Neurons inhibited by CRD (n = 82) were in the area dorsal to the lateral reticular nucleus (LRN). Most neurons excited by CRD (n = 130) were located within or immediately adjacent to the LRN, were excited by noxious heat and/or noxious pinch of at least half the body surface and were called bilateral nociceptive specific (bNS) neurons. bNS neurons had accelerating responses to graded CRD (threshold: 20 +/- 2 mmHg). Ten of twelve bNS neurons tested could be antidromically activated by electrical stimulation of the midline cerebellum. Other neurons excited by CRD (n = 71) had mixed responses to cutaneous stimuli and were generally located in the area dorsal to the LRN. Increases in blood pressure due to intravenous phenylephrine did not significantly alter the spontaneous activity of neurons excited by CRD, but altered spontaneous activity (12 excited, four inhibited) in all neurons tested which were inhibited by CRD. Decreases in blood pressure produced by intravenous nitroprusside produced a reciprocal response in most neurons inhibited by CRD and had a delayed onset (20 30 s after bolus administration) excitatory effect on 21 of 27 units excited by CRD. Combined with other studies, these data suggest a role for neurons within and adjacent to the LRN in the modulation of visceral nociception. They also implicate a role for the cerebellum in visceral nociceptive processing. PMID- 9748556 TI - African elephant myoglobin with an unusual autoxidation behavior: comparison with the H64Q mutant of sperm whale myoglobin. AB - Elephant myoglobins both from Asian and African species have a glutamine in place of the usual distal (E7) histidine at position 64. We have isolated native oxymyoglobin directly from the skeletal muscle of African elephant (Loxodonta africana), and examined the autoxidation rate of oxymyoglobin (MbO2) to metmyoglobin (metMb) as a function of pH in 0.1 M buffer at 25 degreesC. As a result, African elephant MbO2 was found to be equally resistant to autoxidation as sperm whale myoglobin. However, the elephant myoglobin exhibited a distinct rate saturation below pH 6. Kinetic analysis of the pH profiles for the autoxidation rate has disclosed that African elephant MbO2 does not show any proton-catalyzed process, such as the one that can play a dominant role in the autoxidation reaction of sperm whale myoglobin by involving the distal histidine as its catalytic residue. Such a greater stability of African elephant MbO2 at low pH could be explained almost completely by the single H64Q mutation of sperm whale myoglobin. In African elephant aqua-metmyoglobin the Soret band was considerably broadened so as to produce another peak in the pentacoordinate 395 nm region. This unique spectral feature was therefore analyzed to show that the myoglobin is in equilibrium between two species, depending upon the presence or absence of a water molecule at the sixth coordinate position. PMID- 9748557 TI - Cancer predictive value of cytogenetic markers used in occupational health surveillance programs: a report from an ongoing study by the European Study Group on Cytogenetic Biomarkers and Health. AB - The cytogenetic endpoints in peripheral blood lymphocytes: chromosomal aberrations (CA), sister chromatid exchange (SCE) and micronuclei (MN) are established biomarkers of exposure for mutagens or carcinogens in the work environment. However, it is not clear whether these biomarkers also may serve as biomarkers for genotoxic effects which will result in an enhanced cancer risk. In order to assess this problem, Nordic and Italian cohorts were established, and preliminary results from these two studies indicated a predictive value of CA frequency for cancer risk, whereas no such associations were observed for SCE or MN. A collaborative study between the Nordic and Italian research groups, will enable a more thorough evaluation of the cancer predictivity of the cytogenetic endpoints. We here report on the establishment of a joint data base comprising 5271 subjects, examined 1965-1988 for at least one cytogenetic biomarker. Totally, 3540 subjects had been examined for CA, 2702 for SCE and 1496 for MN. These cohorts have been followed-up with respect to subsequent cancer mortality or cancer incidence, and the expected values have been calculated from rates derived from the general populations in each country. Stratified cohort analyses will be performed with respect to the levels of the cytogenetic biomarkers. The importance of potential effect modifiers such as gender, age at test, and time since test, will be evaluated using Poisson regression models. The remaining two potential effect modifiers, occupational exposures and smoking, will be assessed in a case-referent study within the study base. PMID- 9748558 TI - Biochemical and molecular aspects of genetic disorders of bilirubin metabolism. AB - Bilirubin, the oxidative product of heme in mammals, is excreted into the bile after its esterification with glucuronic acid to polar mono- and diconjugated derivatives. The accumulation of unconjugated and conjugated bilirubin in the serum is caused by several types of hereditary disorder. The Crigler-Najjar syndrome is caused by a defect in the gene which encodes bilirubin UDP glucuronosyltransferase (UGT), whereas the Dubin-Johnson syndrome is characterized by a defect in the gene which encodes the canalicular bilirubin conjugate export pump of hepatocytes. Animal models such as the unconjugated hyperbilirubinemic Gunn rat, the conjugated hyperbilirubinemic GY/TR-, and the Eisai hyperbilirubinemic rat, have contributed to the understanding of the molecular basis of hyperbilirubinemia in humans. Elucidation of both the structure of the UGT1 gene complex, and the Mrp2 (cMoat) gene which encodes the canalicular conjugate export pump, has led to a greater understanding of the genetic basis of hyperbilirubinemia. PMID- 9748559 TI - Binding sites for exogenous and endogenous non-competitive inhibitors of the nicotinic acetylcholine receptor. AB - The nicotinic acetylcholine receptor (AChR) is the paradigm of the neurotransmitter-gated ion channel superfamily. The pharmacological behavior of the AChR can be described as three basic processes that progress sequentially. First, the neurotransmitter acetylcholine (ACh) binds the receptor. Next, the intrinsically coupled ion channel opens upon ACh binding with subsequent ion flux activity. Finally, the AChR becomes desensitized, a process where the ion channel becomes closed in the prolonged presence of ACh. The existing equilibrium among these physiologically relevant processes can be perturbed by the pharmacological action of different drugs. In particular, non-competitive inhibitors (NCIs) inhibit the ion flux and enhance the desensitization rate of the AChR. The action of NCIs was studied using several drugs of exogenous origin. These include compounds such as chlorpromazine (CPZ), triphenylmethylphosphonium (TPMP+), the local anesthetics QX-222 and meproadifen, trifluoromethyl-iodophenyldiazirine (TID), phencyclidine (PCP), histrionicotoxin (HTX), quinacrine, and ethidium. In order to understand the mechanism by which NCIs exert their pharmacological properties several laboratories have studied the structural characteristics of their binding sites, including their respective locations on the receptor. One of the main objectives of this review is to discuss all available experimental evidence regarding the specific localization of the binding sites for exogenous NCIs. For example, it is known that the so-called luminal NCIs bind to a series of ring-forming amino acids in the ion channel. Particularly CPZ, TPMP+, QX-222, cembranoids, and PCP bind to the serine, the threonine, and the leucine ring, whereas TID and meproadifen bind to the valine and extracellular rings, respectively. On the other hand, quinacrine and ethidium, termed non-luminal NCIs, bind to sites outside the channel lumen. Specifically, quinacrine binds to a non-annular lipid domain located approximately 7 A from the lipid-water interface and ethidium binds to the vestibule of the AChR in a site located approximately 46 A away from the membrane surface and equidistant from both ACh binding sites. The non-annular lipid domain has been suggested to be located at the intermolecular interfaces of the five AChR subunits and/or at the interstices of the four (M1-M4) transmembrane domains. One of the most important concepts in neurochemistry is that receptor proteins can be modulated by endogenous substances other than their specific agonists. Among membrane-embedded receptors, the AChR is one of the best examples of this behavior. In this regard, the AChR is non-competitively modulated by diverse molecules such as lipids (fatty acids and steroids), the neuropeptide substance P, and the neurotransmitter 5 hydroxytryptamine (5-HT). It is important to take into account that the above mentioned modulation is produced through a direct binding of these endogenous molecules to the AChR. Since this is a physiologically relevant issue, it is useful to elucidate the structural components of the binding site for each endogenous NCI. In this regard, another important aim of this work is to review all available information related to the specific localization of the binding sites for endogenous NCIs. For example, it is known that both neurotransmitters substance P and 5-HT bind to the lumen of the ion channel. Particularly, the locus for substance P is found in the deltaM2 domain, whereas the binding site for 5-HT and related compounds is putatively located on both the serine and the threonine ring. Instead, fatty acid and steroid molecules bind to non-luminal sites. More specifically, fatty acids may bind to the belt surrounding the intramembranous perimeter of the AChR, namely the annular lipid domain, and/or to the high-affinity quinacrine site which is located at a non-annular lipid domain. Additionally, steroids may bind to a site located on the extracellular hydrophi PMID- 9748560 TI - Clinical applications of anticancer drugs targeted to topoisomerase II. AB - Agents which 'poison' the enzyme topoisomerase II, have proven to be useful drugs for cancer treatment. Six antineoplastic drugs, which target topoisomerase II (doxorubicin, daunorubicin, idarubicin, mitoxantrone, etoposide and teniposide) are currently approved for clinical use in the United States. In this paper, the strategies and goals of cancer chemotherapy are summarized for the non-clinician. The use, pharmacology and toxicity of each of the six currently approved topoisomerase II inhibiting agents are reviewed. PMID- 9748561 TI - Effect of neutrophil depletion in acute cerebritis. AB - Inhibition of the host's neutrophil response has been proposed as one means to reduce tissue damage in acute inflammation. If this approach can be applied in acute central nervous system (CNS) infection, the long-term morbidity, which occurs in CNS infection, might be reduced. Previous studies in models of CNS infection yielded conflicting results whether neutrophil depletion might be protective. To determine whether neutrophil depletion reduces tissue necrosis and cerebrovascular injury in experimental bacterial cerebritis, we depleted circulating neutrophils with an IgM monoclonal antibody, RP3, given after the start of the infection. RP3 treatment successfully depleted circulating neutrophils and reduced the extent of neutrophil influx into the cerebritis region. The extent of tissue necrosis, measured histologically, and the regional increase of blood-brain barrier (BBB) permeability were not inhibited by neutrophil depletion, and in animals treated with RP3 alone, the extent of tissue necrosis and BBB permeability tended to be larger than in S. aureus inoculated controls. We conclude that host neutrophils do not add to the tissue and cerebrovascular damage created by the intracerebral inoculation of a pathogenic bacteria, and the neutrophils serve to diminish local damage in the setting of a cerebritis. PMID- 9748562 TI - Evidence that the influence of ganglion cell axons on astrocyte morphology is mediated by action spike activity during development. AB - In many mammal retinas, the morphology of astrocytes is strongly influenced by nearby axons of ganglion cells. Astrocyte processes stretch along the axons, fine extensions of the processes contact node-like specialisation of the axon membrane and the morphology of the adult astrocyte is strongly determined by this relationship. The mechanism which attracts astrocyte processes to contact specific regions of the axon membrane is not known however. This study presents evidence that in the neonatal cat blocking the impulse activity of ganglion cells with the Na+-channel blocker tetrodotoxin (TTX) leads to a loss of the axon related morphology of astrocytes. The morphological change induced in astrocytes by TTX was greater in younger animals and could not be detected in the adult. Conversely, if the TTX block was maintained for 4 postnatal weeks the changes induced in astrocytes persisted at least to 13 weeks. The TTX-induced loss of axon-related morphology in astrocytes suggests that the signal by which axons attract astrocyte processes to contact the axonal membrane in ways which modify astrocyte morphology is released by action spike activity during development. PMID- 9748563 TI - Kinetic study of chemoselective acylation of amino-alditol by immobilized lipase in organic solvent: effect of substrate ionization. AB - The kinetics of the lipase-catalyzed synthesis of oleoyl-N-methylglucamide and 6 O-oleoyl-N-methylglucamine in organic systems were investigated. We have shown that in apolar media, the ionic state of substrates and the ionic state of enzyme microenvironment play an important role in immobilized Candida antarctica lipase activity and chemoselectivity of the reaction. In order to define the optimal conditions of the reaction, to obtain the highest initial rate for amide formation, the influence of acid/N-methylglucamine molar ratio is studied. This ratio determines the protonation states of substrates and of ionizable groups of catalytic site, on which the enzyme activity is dependent. To confirm our hypothesis, we have added to the medium a non-reactive base which is not a substrate of the enzyme. We observed that when the acid/base ratio is higher than 1, the initial rate of ester synthesis increases whereas that of amide synthesis decreases. On the opposite, when the acid/base ratio is lower than 1, the initial rate of amide synthesis becomes preponderant. PMID- 9748564 TI - Transgenic systems in studies on genotoxicity of alkylating agents: critical lesions, thresholds and defense mechanisms. AB - Transgenic systems, both cell lines and mice with gain or loss of function, are being used in order to modulate the expression of DNA repair proteins, thus allowing to assess their contribution to the defense against genotoxic mutagens and carcinogens. In this review, questions have been addressed concerning the use of transgenic systems in elucidating critical primary DNA lesions, their conversion into genotoxic endpoints, low-dose effects, and the relative contribution of individual cellular functions in defense. It has been shown that the repair protein alkyltransferase (MGMT) is decisive for protection against methylating and chloroethylating compounds. Protection pertains also to tumor formation, as revealed by the response of MGMT transgenic and knockout mice. Overexpression of genes involved in base excision repair (N-methylpurine-DNA glycosylase, apurinic endonuclease, DNA polymerase beta) is in most cases not beneficial in increasing the protection level, whereas their down-modulation or inactivation increases cellular sensitivity. This indicates that non-repaired base N-alkylation lesions and/or repair intermediates possess genotoxic potential. Modulation of mismatch repair and poly(ADP)ribosyl transferase has also been shown to affect the cellular response to alkylating agents. Furthermore, the role of Fos, Jun and p53 in cellular defense against alkylating mutagens is discussed. PMID- 9748565 TI - Mechanism of factor IXa inhibition by antithrombin in the presence of unfractionated and low molecular weight heparins and fucoidan. AB - Heparin exerts its anticoagulant activity by catalysing the inhibition of coagulation proteases by antithrombin (AT). Its main target is thrombin but it also catalyses the inhibition of the other serine-proteases of the coagulation cascade, such as factor IXa (fIXa). The aim of this study was to compare the catalysis of inhibition of blood fIXa by antithrombin in the presence of several sulfated polysaccharides with anticoagulant activity, i.e. heparin, three widely used in therapeutics low molecular weight heparins (LMWH) and fucoidan. Plots of the second-order rate constants of the fIXa-antithrombin reaction vs. the concentration of added heparin and LMWH are bell-shaped and fit the kinetic model established for thrombin-antithrombin reaction by Jordan R., Beeler D., Rosenberg R. (1979) J. Biol. Chem., 254, 2902-2913. In the ascending branch, the catalyst (C) binds quickly to the inhibitor (I) to form a catalyst-inhibitor (CI) complex which is more reactive towards the enzyme (E) than the free inhibitor, leading to the formation of an inactive enzyme-inhibitor complex (EI) and the release of free catalyst, in a rate-limiting second step. After a maximum corresponding to an optimal catalyst concentration, the decrease in the reaction rate was in keeping with the formation of a catalyst-enzyme (CE) complex, whose inactivation by the CI complex was slower than that of the free enzyme. Maximum second-order rate constants for the inhibition of fIXa by AT were 105, 6.8, 12.24 and 22 microM-1 min-1 with heparin, Enoxaparin, Fraxiparin and Fragmin, respectively, leading to 3500-, 225-, 405- and 728-fold increases in the inhibition rate in the absence of polysaccharide, respectively. Fucoidan yielded 23-fold increase in the fIXa-antithrombin interaction rate. The kinetic profiles obtained with this polysaccharide exhibited ascending branch which correlated well with the kinetic model based on the formation of binary complexes (CI or CE). Fucoidan was covalently conjugated with a fluorescent probe (DTAF) and used in conjunction with fluorescence anisotropy to follow its binding to antithrombin, heparin cofactor II (HCII), thrombin and fIXa. The binding of fucoidan to these proteins occurred with low affinities when compared to heparin and LMWH. Fucoidan had higher affinity for the inhibitor HCII compared to antithrombin and enzymes. These data suggest that binding of heparins and fucoidan to the inhibitor (CI) is required for the polysaccharide-dependent enhancement in the rate of neutralization of the enzyme by the inhibitor. PMID- 9748566 TI - Association between the amygdala and nucleus of the solitary tract in mu-opioid induced feeding in the rat. AB - The central nucleus of the amygdala (CNA) and the nucleus of the solitary tract (NTS) are important in the regulation of ingestive behavior. We evaluated whether opioid-opioid signaling between the CNA and rostral NTS (rNTS) affect feeding behavior. To test this, rats were doubly cannulated with one cannula placed in the rNTS and one cannula in the CNA, allowing for co-administration of an opioid agonist into one site and an opioid antagonist into the other. Tyr-D-Ala-Gly-(me) Phe-Gly-ol (DAMGO) (2 nmol) injected into the CNA (CNA DAMGO) increased feeding more than two-fold compared to the vehicle-injected rats. This increase in food intake was blocked when doses of 26.5 and 79 nmol of naltrexone (NTX) were injected into the rNTs. In the reverse situation, rNTS DAMGO increased food intake above control levels, and CNA NTX blocked DAMGO-induced feeding when administrated in doses of 26.5 and 79 nmol. This suggests that a bi-directional opioid-opioid signaling pathway exists between the CNA and the rNTS which influences feeding via mu-opioid receptors. PMID- 9748567 TI - The role of alternative splicing in regulating agrin binding to muscle cells. AB - The binding of agrin to the muscle cell surface can induce radical changes in the topography and physiology of the cell membrane, resulting in the organization of postsynaptic components opposite the nerve terminal. Alternative splicing of agrin mRNA yields several isoforms, which vary in their cellular expression, developmental profile, and acetylcholine receptor (AChR) clustering activity. Neurons and muscle cells express several of these agrin isoforms. To address the role of alternative splicing in regulating agrin's function, we compared the effects of splicing at the y and z sites of agrin (denoted 'Agy,z'). Agrin isoforms bound differently to the myotube surface: Ag0,0 and Ag4,0 showed much higher levels of binding than Ag4,8. The artificial splice form Ag0,8 showed binding levels similar to Ag4,8. Visualization of the bound agrin after an acute incubation revealed that each isoform associated with the cell surface in a distinct pattern. These binding patterns changed following stimulation of the myotubes with Ag4,8 for 4 h (which induces the clustering of AChRs). Ag4,8 binding sites were concentrated at >90% of the induced AChR clusters, while those for Ag4,0, Ag0,8, and Ag0,0 were enriched at 70%, 50% and 25%, respectively. Together, these observations indicate that alternatively spliced forms of agrin recognize at least partially non-overlapping populations of binding sites on the cell surface, and that the eight amino acid insert is the dominant factor influencing the level of the agrin binding to the cell surface. Further, some of these populations redistribute to AChR clusters upon agrin stimulation. PMID- 9748568 TI - DNA sequence selectivity of topoisomerases and topoisomerase poisons. AB - Chemical agents able to interfere with DNA topoisomerases are widespread in nature, and some of them have clinical efficacy as antitumor or antibacterial drugs. Drugs which have as a target DNA topoisomerases could be divided into two categories: poisons and catalytic inhibitors. Classical topoisomerase poisons stimulate cleavage in a sequence-selective manner, yielding drug-specific cleavage intensity pattern. The mechanisms of drug interaction with DNA topoisomerases, the DNA sequence selectivity of the action of topoisomerase II poisons and the identification of structural determinants of their activity have suggested that topoisomerase II poisons may fit into a specific pharmacophore, constituted by a planar ring system with DNA intercalation or intercalation-like properties, and protruding side chains interfering with the protein side of the covalent enzyme-DNA complex. The complete definition of the diverse pharmacophores of topoisomerase II poisons will certainly be of value for the design of new agents directed to specific genomic sites, and more effective in the treatment of human cancer. PMID- 9748569 TI - A simple screening test for variant transthyretins associated with familial transthyretin amyloidosis using isoelectric focusing. AB - Variant forms of the plasma protein transthyretin (TTR) are associated with the most frequently occurring type of familial systemic amyloidosis. Organ system involvement in transthyretin type amyloidosis (ATTR) is often similar to that which occurs in light chain amyloid disease (AL). The proper diagnosis of ATTR is important since treatment (liver transplantation) differs from that in AL (chemotherapy). We present a two-step test to screen sera for variant TTRs using non-denaturing gel electrophoresis performed in 7.5% acrylamide (PAGE) followed by isoelectric focusing (IEF) between pH 4.0 and 7.0 in 2.5 M urea. Serum samples from 110 patients with amyloidosis and their relatives were tested using this IEF technique and compared to genetic mutation results. Sera from patients with ATTR who underwent liver transplantation were also examined prior to and following surgery. IEF analysis showed the presence of both wild-type and variant TTR in 74 of the 110 serum samples tested. Genomic DNA from peripheral blood was used to identify TTR gene mutations in 77 of the 110 patients. Fifteen variants including Val122Ile, preponderant in the African-American population, could be demonstrated by IEF. The sensitivity of IEF was 96% (74/77) and the specificity was 100% (33/33). The predictive values for a positive or negative result were 100% (74/74) and 92% (33/36), respectively. There were no false-positive results and 4% (3/77) false-negative results. In sera from patients with ATTR who underwent liver transplantation, variant TTR was detected by IEF before, but not after, surgery. A simple, accurate, sensitive method is presented as a useful screening test for variant transthyretins associated with ATTR. PMID- 9748570 TI - Cyclooxygenase 2 mRNA expression in rat brain after peripheral injection of lipopolysaccharide. AB - Inducible cyclooxygenase 2 (COX 2) converts arachidonic acid to prostaglandins, which are thought to mediate various peripheral lipopolysaccharide (LPS)-induced central effects, including generation of fever and activation of the hypothalamic pituitary-adrenal axis. To localize prostaglandin production in the brain following peripheral LPS administration, COX 2 mRNA expression was examined by in situ hybridization histochemistry in rats injected intraperitoneally (i.p.) or intravenously (i.v.) with various doses of LPS or saline. Constitutive expression of COX 2 mRNA was found in neurons of cortex, hippocampus, and amygdala, but not in cells of the blood vessels. COX 2 mRNA levels were not altered in saline injected animals as compared to non-injected controls. In LPS-injected animals, no consistent changes of neuronal COX 2 mRNA expression were observed. COX 2 mRNA expression appeared ex novo at 0.5-h post-injection in cells closely associated with blood vessels, however, ex novo labeling of the number of labeled cells increased to a peak at 2 h and subsided gradually to basal levels by 24 h. Initially, labeling was observed in cells comprising major surface-lying blood vessels and meninges. Later, vascular and perivascular cells associated with smaller penetrating blood vessels were labeled. This pattern of COX 2 mRNA induction is independent of the route and dose of the LPS injection. The induced COX 2 mRNA producing cells are identified as endothelial and leptomeningeal cells. Changes in COX 2 mRNA expression were not observed in circumventricular organs. These results suggest that peripheral LPS induces a rapid increase in COX 2 production throughout the vasculatures of the brain, which could affect the neuronal activity of widespread brain regions by elevating the levels of prostaglandins. PMID- 9748571 TI - Developmental characteristics of neuronal nitric oxide synthase (nNOS) immunoreactive neurons in fetal to adolescent human brains. AB - The developmental characteristics of the neuronal nitric oxide synthase (nNOS) immunoreactive neurons in the human brain were studied. In the frontal lobe, nNOS immunoreactive cells appeared as early as 18 gestational weeks (GW) in the subcortical plate and then increased predominantly in the subcortical white matter during the fetal period, while weakly immunoreactive neurons were found in the cortical II-IV layers after 26 GW. In the basal ganglia, immunoreactive neurons could be detected in the striatum as early as 13 GW, and then showed a transient increase with peaks at 23-24 GW and 33-36 GW in the putamen and caudate nucleus, respectively. In the cerebellum, immunoreactivity was detected in the Purkinje and basket cells after 23 GW and 31 GW, respectively. The immunoreactivity of internal granule cells was constantly weak. In the brain stem, constant and intense immunoreactive neurons were found in the central gray, pedunculopontine tegmental nucleus, solitary tract nucleus, and lateral reticular nucleus. The immunoreactivity in the neurons of the pontine nucleus and inferior olivary nucleus was transiently increased, with peaks at 38-40 GW and 23-24 GW, respectively. This characteristic nNOS development suggests that transient nNOS hyperproduction may contribute to neuron maturation as well as vulnerability in each period and region, and NO may play an important role in the basic development of human brain functions. PMID- 9748572 TI - Mutagenic activity of carcinogens detected in transgenic rodent mutagenicity assays at dose levels used in chronic rodent cancer bioassays. AB - Data on transgenic rodent mutagenicity of five human carcinogens were summarised and compared with the results from rodent carcinogenicity studies. Four out of five carcinogens showed mutagenic activity already at daily dose levels which induced cancer in long-term rodent bioassays in at least one target tissue of carcinogenesis. In several of these studies, even single dose applications were sufficient to significantly increase the mutation frequency in vivo. Other genotoxic carcinogens required application of multiple dosing at dose-levels used in rodent cancer bioassays to show their in vivo mutagenicity. A rodent respiratory tract carcinogen, 1,2-dibromoethane (DBE), following inhalation exposure, displayed no mutagenic activity, neither in lung nor in nasal mucosa, at a single 2-h exposure to 30 ppm, which is below the highest concentration used in a NTP cancer bioassay. In contrast, after multiple treatment for 10 days at the same daily doses, a significant increase of the mutation frequency in nasal mucosa was apparent. We conclude, that especially when studying new chemicals in these transgenic rodent mutation assays, a multiple dosing protocol should be preferred. For dose selection, the same criteria could be applied as for chronic rodent bioassays. PMID- 9748573 TI - Inhibition of post-ischemic reperfusion injury of the kidney by diamine oxidase. AB - To elucidate the role of histamine in the pathogenesis of post-ischemic reperfusion injury of tissues, the effect of diamine oxidase (DAO) was studied on the changes in renal functions induced by 30 min occlusion followed by reperfusion of the renal vessels of unilaterally nephrectomized rats. Kinetic analysis using radiolabeled albumin revealed that vascular permeability of the kidney increased markedly after reperfusion. Although the intensity of neutrophil dependent chemiluminescence of the blood remained unchanged during the occlusion, it increased significantly after reperfusion. Histological examination revealed a marked degeneration of glomeruli and proximal tubules in the reperfused kidney. Transtubular transport of phenolsulfophthalein (PSP) decreased markedly after reperfusion with concomitant increase in plasma levels of creatinine. Intravenously administered DAO markedly inhibited the reperfusion-induced increase in vascular permeability, preserved the structure of the kidney and normalized the rate of clearance of PSP and creatinine. Combined use of diphenylhydramine and ranitidine also inhibited the reperfusion injury of the kidney. These results suggested that histamine and its receptors might play critical roles in post-ischemic reperfusion injury of the kidney. PMID- 9748574 TI - Acetonitrile-protein interactions: amino acid solubility and preferential solvation. AB - The solubility of amino acids and the preferential solvent interaction of hen-egg lysozyme in acetonitrile (AN)-water mixtures (<60 w/v% AN) were investigated by means of densimetric and refractometric methods at 25 degreesC. The free energy of transfer from water to aqueous AN was negative for most nonpolar side-chains of amino acids and positive for the peptide group, the extent being comparable to those for methanol and ethanol systems. Addition of AN to an aqueous solvent was thus suggested to weaken the hydrophobic interaction and to enhance the peptide peptide hydrogen bond therein leading to the denaturation of proteins. A parallel examination by circular dichroism confirmed that the conformation of lysozyme (pH 3) remains native in aqueous AN up to 40% but changes to the helix-rich form at higher AN concentrations. At all solvent compositions up to 50% AN (pH 3), however, lysozyme was preferentially hydrated probably due to a local salting-out of the AN molecules from the charges on the protein surface, indicating the increase of the chemical potential of the protein. These results are discussed in relation to the role of AN as an eluting organic solvent in reverse-phase chromatography. PMID- 9748576 TI - Leukemia inhibitory factor and NGF regulate signal transducers and activators of transcription activation in sympathetic ganglia: convergence of cytokine- and neurotrophin-signaling pathways. AB - We have used the response of the superior cervical ganglia (SCG) to axotomy to investigate interactions between neuropoietic cytokines and neurotrophins. Postganglionic sympathetic axotomy leads to a prolonged leukemia inhibitory factor (LIF)-dependent activation of signal transducers and activators of transcription (STAT) factors. To study regulation of LIF-dependent activation of STAT proteins and to mimic the loss of target-derived NGF resulting from postganglionic axotomy in vivo, SCG were explanted into media lacking NGF and activation of STAT proteins was assessed by electrophoretic mobility shift assay. Like postganglionic axotomy in vivo. STAT proteins were activated for up to 8 days after explantation of SCG in vitro. SCG cultured in the presence of NGF showed decreased STAT binding when compared to ganglia cultured in NGF-free media. This inhibition of STAT activation by NGF was only present in ganglia cultured for more than 5 days and was mimicked by brain-derived neurotrophic factor (BDNF). The serine kinase inhibitor H7 augmented the increase of STAT binding produced by explantation, suggesting the presence of a labile repressor of STAT activation in the SCG. These data indicated that the neuropoietic cytokine-signaling pathway interacts with neurotrophin and H7-sensitive-signaling pathways to regulate activation of STAT proteins in sympathetic neurons. Moreover, these data suggest that one of the mechanisms leading to prolonged activation of STAT proteins after postganglionic axotomy in vivo is loss of target-derived neurotrophins. PMID- 9748575 TI - Cell death induced by topoisomerase-targeted drugs: more questions than answers. AB - Chemotherapeutic agents that target topoisomerase I and II set into motion a series of biochemical changes that culminate in cell death, but only under some conditions. The realization that stabilization of covalent topoisomerase-DNA complexes is not sufficient to insure cell death has prompted investigators to examine various aspects of the drug-induced death process itself. Several discrete steps along this pathway have been identified, including (a) the processing of stabilized cleavage complexes into frank DNA strand breaks; (b) sensing of the DNA damage, leading to activation of stress-associated signaling pathways and cell cycle arrest; and (c) activation of a preexisting group of enzymes and enzyme precursors, typified by the cysteine-dependent aspartate directed proteases (caspases), that catalyze the relatively orderly biochemical cascade of terminal events known as apoptosis. The present review discusses the evidence that these steps occur after treatment with etoposide or camptothecin, the two prototypic topoisomerase poisons that are commonly studied. As in any emerging area, a large number of questions remain to be answered about the process of cell death induced by topoisomerase-directed drugs. PMID- 9748577 TI - Low dose effects of chemicals as assessed by the flow cytometric in vivo micronucleus assay. AB - Using flow cytometric automation of the mouse in vivo, micronucleus assay increases the sensitivity of the test. This is achieved through a very large increase in the number of cells scored, by a factor of 100x, which in turn greatly reduces the sampling error. With this method, dose-response relationships of in vivo micronucleus induction for four model agents mitomycin C (MMC), diepoxybutane (DEB), cyclophosphamide (CPA), and colchicine (COL) were studied at low dose levels. For the three clastogens MMC, DEB and CPA, linear dose-response relationships were found over the dose ranges studied, even in the very low dose region (defined as the dose region where the frequency of micronucleated erythrocytes is less than twice the baseline frequency). This is consistent with the view that no threshold should exist for genotoxic agents which target DNA. For COL a dose range was found, in which the frequency of micronucleated erythrocytes did not increase with dose, possibly indicating an in vivo threshold. The flow cytometric in vivo micronucleus assay represents one possibility for in vivo low dose-response studies. PMID- 9748578 TI - Identification of novel genes that are differentially expressed in human colorectal carcinoma. AB - By using mRNA differential display technology, we have identified three cDNA clones, myl 3, myl 4, and myl 6, to show significant changes in expression between human colorectal tumor and paired normal tissue. Northern blot analysis indicated that clone myl 3 expression was elevated in normal tissue, and clone myl 4 expression was elevated in tumor tissue. Nucleotide sequence analysis revealed that clones myl 3 and myl 4 have not been previously identified. However, clone myl 6 appears to be the human homolog of the 3' end region of tissue inhibitor of metalloproteinase 3 (TIMP-3). Northern blot analysis showed that a 2.5 kb TIMP-3 transcript was expressed at a much higher level in normal tissue than the colorectal tumor. PMID- 9748579 TI - Postnatal development of neuron number and connections in the suprachiasmatic nucleus of the hamster. AB - We had previously found a ca. 30% cell death during the prenatal ontogeny of the suprachiasmatic nucleus (SCN) of lambs. The period of neuron death was preceded by the establishment of the retinohypothalamic connections, a major input to this nucleus that allows the entrainment to light of the circadian rhythms generated by the SCN. The present study determined the temporal relationship between the period of ontogenetic neuron death and the establishment of the principal afferent and efferent connections of the SCN in hamsters. We found that during the first 3 postnatal days the SCN volume increases mainly by the addition of cells. After a peak 6140 neurons on each side during the third postnatal day, the SCN underwent an acute decrease of about 40% in neuron number, which led to the final adult complement of neurons, estimated in 3400 neurons per nucleus. The connections of the SCN were studied by placing DiI crystals either into the optic nerve, or into the SCN of brains fixed at different ages. We found, in agreement with previous studies, that retinal axons can be detected after the fifth postnatal day, that is, after the large decrease in neuron number. As for the SCN efferents, they began to invade the appropriate targets during the second postnatal day, followed by a large increase in the density of these efferent projection in the subsequent days. In conclusion, the massive neuronal death in the SCN was preceded by the formation of efferent connections, and followed by the formation of the retinohypothalamic tract. PMID- 9748580 TI - Neural control of left ventricular contractility in the dog heart: synaptic interactions of negative inotropic vagal preganglionic neurons in the nucleus ambiguus with tyrosine hydroxylase immunoreactive terminals. AB - Recent physiological evidence indicates that vagal postganglionic control of left ventricular contractility is mediated by neurons found in a ventricular epicardial fat pad ganglion. In the dog this region has been referred to as the cranial medial ventricular (CMV) ganglion [J.L. Ardell, Structure and function of mammalian intrinsic cardiac neurons, in: J.A. Armour, J.L. Ardell (Eds.). Neurocardiology, Oxford Univ. Press, New York, 1994, pp. 95-114; B.X. Yuan, J.L. Ardell, D.A. Hopkins, A.M. Losier, J.A. Armour, Gross and microscopic anatomy of the canine intrinsic cardiac nervous system, Anat. Rec., 239 (1994) 75-87]. Since activation of the vagal neuronal input to the CMV ganglion reduces left ventricular contractility without influencing cardiac rate or AV conduction, this ganglion contains a functionally selective pool of negative inotropic parasympathetic postganglionic neurons. In the present report we have defined the light microscopic distribution of preganglionic negative inotropic neurons in the CNS which are retrogradely labeled from the CMV ganglion. Some tissues were also processed for the simultaneous immunocytochemical visualization of tyrosine hydroxylase (TH: a marker for catecholaminergic neurons) and examined with both light microscopic and electron microscopic methods. Histochemically visualized neurons were observed in a long slender column in the ventrolateral nucleus ambiguus (NA-VL). The greatest number of retrogradely labeled neurons were observed just rostral to the level of the area postrema. TH perikarya and dendrites were commonly observed interspersed with vagal motoneurons in the NA VL. TH nerve terminals formed axo-dendritic synapses upon negative inotropic vagal motoneurons, however the origin of these terminals remains to be determined. We conclude that synaptic interactions exist which would permit the parasympathetic preganglionic vagal control of left ventricular contractility to be modulated monosynaptically by catecholaminergic afferents to the NA-VL. PMID- 9748581 TI - Changes in cannabinoid receptor binding and mRNA levels in several brain regions of aged rats. AB - We have recently found that cannabinoid receptor binding and gene expression markedly decreased in extrapyramidal structures of aged rats. The present study was designed to analyze the possible existence of similar aging-induced changes in cannabinoid receptor binding and gene expression in brain regions other than extrapyramidal areas, but that also contain a significant population of cannabinoid receptors, such as the cerebellum, hippocampal structures, limbic and hypothalamic nuclei, the cerebral cortex and others. To this end, we analyzed cannabinoid receptor binding, using autoradiography, and cannabinoid receptor mRNA levels, using in situ hybridization, in slide-mounted brain sections obtained from young (3 month old) and aged (> 2 year old) rats. Results were as follows. In the cerebellum, aged rats exhibited a marked decrease in cannabinoid receptor binding in the molecular layer (-33.3%), although accompanied by no changes in mRNA levels in the granular layer. In the cerebral cortex, a small, although statistically significant, decrease in binding was found in the deep layer (VI) (-18.3%) of aged rats, whereas no changes were found in the superficial layer (I). As in the case of the cerebellum, mRNA levels did not change in the cerebral cortex layers (II-III and V-VI). The different regions of the Ammon's horn of the hippocampus exhibited similar cannabinoid receptor binding levels in aged and young rats. Interestingly, mRNA levels decreased in aged rats to a small, but statistically significant, extent (CA1: -26.1%; CA2: 21.6%; CA3: -14.4%). This was also seen in another hippocampal structure, the dentate gyrus (-14.6%), although in this region binding levels increased in aged rats (+28.4%). Two hypothalamic structures, the arcuate nucleus and the ventromedial hypothalamic nucleus, exhibited decreased cannabinoid receptor binding in aged rats (-31.1% and -30.3%, respectively), but this was not seen in the medial preoptic area. This was accompanied by no changes in mRNA levels in the ventromedial hypothalamic nucleus. In the limbic structures, aged rats exhibited similar binding levels to young rats. This was seen in the nucleus accumbens, septum nuclei and basolateral amygdaloid nucleus. However, mRNA levels slightly decreased in the basolateral amygdaloid nucleus (-13.4%), whereas they were not altered in the septum nuclei. Finally, other brain structures, such as the central gray substance and the brainstem, exhibited similar binding levels in aged and young rats. However, it is important to note that mRNA levels increased significantly (+211.2%) in the brainstem of aged rats, an area where the levels of binding and mRNA were very low in young rats. This marked increase may be related to an increase in the presence of glial elements in this region, as revealed by the increase in the immunoreactivity for glial fibrillary acidic protein observed in the brainstem of aged rats as compared to young animals. In summary, senescence was associated with changes in cannabinoid receptors in the cerebellum, the cerebral cortex, limbic and hypothalamic structures, the hippocampus and other brain regions. However, the changes observed (i) were not as marked and relevant as those early reported in extrapyramidal areas, and (ii) exhibited regional differences that might be attributed to the different roles played by these receptors in each region. Of particular relevance by their magnitude were the aging-induced decrease in binding found in the cerebellum and the hypothalamus, and the increase in mRNA levels observed in the brainstem. The latter might be related to an increase in the presence of glial cells which might contain cannabinoid receptor mRNA. PMID- 9748582 TI - Equine CRISP-3: primary structure and expression in the male genital tract. AB - Although originally described in the male rodent genital tract, cysteine-rich secretory proteins (CRISPs) are expressed in a variety of mammalian tissue and cell types. The proteins of the male genital tract have been observed associated to spermatozoa and are believed to play a role in mammalian fertilization. Here we describe the identification and primary structure of the first equine member of the CRISP family. Equine CRISP-3 is transcribed and expressed in the stallion salivary gland, in the ampulla and the seminal vesicle. It displays all 16 conserved cysteine residues and shows 82% homology to human and 78% to guinea pig CRISP-2 (AA1, TPX 1) and 77% to human CRISP-3. In contrast to other mammalia, in the horse CRISP-3 is synthesized in great amounts in the accessory sexual glands, ampulla and seminal vesicle, thus allowing the isolation of equine CRISP-3 in amounts suitable for biochemical, physiological and structural studies from stallion seminal plasma. PMID- 9748583 TI - Mutation analysis using the restriction site mutation (RSM) assay. AB - The restriction site mutation (RSM) assay (see Steingrimsdottir et al. [H. Steingrimsdottir, D. Beare, J. Cole, J.F.M. Leal, T. Kostic, J. Lopez-Barea, G. Dorado, A.R. Lehmann, Development of new molecular procedures for the detection of genetic alteration in man, Mutat. Res. 353 (1996) pp. 109-121] for a review) has been developed as a genotypic mutation detection system capable of identifying mutations occurring in restriction enzyme sites of genomic DNA. Here we will report the steps taken to overcome some of the initial problems of the assay, namely the lack of quantitative data and limited sensitivity, the aim being to achieve a methodology suitable for the study of low dose chemical exposures. Quantitative data was achieved in the RSM assay by the inclusion of an internal standard molecule in the PCR amplification stage, thus allowing the calculation of both spontaneous and induced mutation frequencies. The sensitivity of the assay was increased through the discovery that intron sequences of genomic DNA accumulated more mutations in vivo compared to the exons, presumably due to differential selective pressure within genes [G.J.S. Jenkins, I.deG. Mitchell, J.M. Parry, Enhanced restriction site mutation (RSM) analysis of 1, 2 dimethylhydrazine-induced mutations, using endogenous p53 intron sequences, Mutagenesis 12 (1997) pp. 117-123]. This increased sensitivity was examined by applying the RSM assay to analyse the persistence of N-ethyl-N-nitrosourea (ENU) induced mutations in mice testes. Germ line mutations were sought in testes DNA 3, 10 and 100 days after ENU treatment. Mutations were detected in exons and especially intron regions, the intron mutations were more persistent, still being detected 100 days post-chemical treatment. Assignment of these mutations as ENU induced was complicated in some cases where the spontaneous mutation level was high. This theme of mutation persistence was further investigated by studying the presence of 4-nitroquinoline-1-oxide (4-NQO)-induced DNA mutations in vitro. This study also analysed the relationship between DNA adduct formation and DNA mutation induction by the concurrent RSM analysis and 32P post-labelling analysis of 4-NQO treated human fibroblasts. The results demonstrated that early DNA mutations detected 4 days post-treatment by the RSM assay were probably ex vivo mutations induced by Taq polymerase misincorporation of 4-NQO adducted DNA, due to the maximum levels of 4-NQO adducts being present at this time point. A later mutational peak, after the adduct level had declined, was assumed to be due to DNA sequence changes produced in the fibroblasts by the in vivo processing of DNA adducts. PMID- 9748584 TI - Mutagenic properties of topoisomerase-targeted drugs. AB - Topoisomerases maintain DNA structure by relieving torsional stress occurring in DNA during transcription, replication and cell division. Topoisomerases are of two main types, causing transient breaks in one (type I) or both (type II) and strands of DNA, and a number of clinical anticancer drugs are thought to act by inhibiting religation of these transient breaks. Topoisomerase II appears to have a close association with the SMC (stable maintenance of chromosomes) family of proteins involved in organisation of the chromatin in a series of loops on the proteinaceous chromosomal scaffold. Inhibition of topoisomerase II function can result in deletions of such loops, probably mediated by reciprocal exchange of topoisomerase subunits. Disruption of topoisomerase I and/or II function during DNA replication results in smaller DNA deletions and other mutations, probably arising from non-homologous recombination. Inhibition of topoisomerase II action during mitosis and meiosis can cause incomplete separation of chromatids and chromosomes, with the consequent production of genomic mutations. Topoisomerase mediated mutagenicity is important because it can lead not only to drug resistance but also to drug-induced secondary cancers. Mutagenicity of topoisomerase-directed agents has been underestimated in the past, since these drugs are not usually capable of reacting covalently with DNA and usually have low mutagenicity in microbial assays. PMID- 9748585 TI - Advanced glycation end products induce crosslinking of collagen in vitro. AB - We have investigated the effect of advanced glycation end products (AGEs) on the crosslinking of collagen. The potential pathological significance of AGEs and the altered metabolism of ascorbic acid (ASA) in diabetes have prompted us to investigate the role of ASA in the crosslinking and advanced glycation of collagen. Rat tail tendons were incubated with ASA and dehydroascorbic acid (DHA) under physiological conditions of temperature and pH, and the crosslinking and the level of AGEs were analyzed. Analysis of crosslinking was conducted by pepsin solubility and cyanogen bromide digestion. Level of AGEs was estimated by enzyme linked immunosorbent assay (ELISA) using antibodies raised against AGE ribonuclease. It was noted that ASA and DHA induced crosslinking of collagen and stimulated the formation of AGEs. It was also noted that these pathways were dependent on oxidative conditions. Similarly incubation of collagen with AGEs, prepared by the in vitro incubation of bovine serum albumin (BSA) with glucose, also resulted in increased crosslinking. The extent of crosslinking was dependent on the duration of incubation. The novel finding of this study, which is in contrast to the earlier reports on glucose-induced crosslinking of collagen, was that AGEs-induced crosslinking of collagen was not inhibited by radical scavengers and the metal chelator. EDTA, whereas glucose-induced crosslinking of collagen was almost completely prevented by free radical scavengers. The increased fluorescence intensity observed in collagen incubated with AGEs was also not prevented by radical scavengers. Estimation of AGEs by ELISA revealed an increased accumulation of AGEs in collagen incubated with AGE-BSA. The inhibitory effect of aminoguanidine and aspirin on AGEs-induced modification of collagen, strongly suggests that the amino-carbonyl interaction between AGEs and collagen may play a key role in the crosslinking process. The results obtained in this study indicate that soluble AGEs can directly induce crosslinking of collagen and this process is independent of oxidative conditions. From these results it may be hypothesized that glucose, under oxidative conditions, reacts with proteins to form potentially reactive end products called AGEs. These AGEs, once formed, could induce crosslinking of collagen even in the absence of both glucose and oxygen. PMID- 9748587 TI - Inhibition of acetylcholinesterase by an alkylpyridinium polymer from the marine sponge, Reniera sarai. AB - Large polymeric 3-alkylpyridinium salts have been isolated from the marine sponge Reniera sarai. They are composed of N-butyl(3-butylpyridinium) repeating subunits, polymerized head-to-tail, and exist as a mixture of two main polymers with molecular weights without counterion of about 5520 and 18900. The monomer analogue of the inhibitor, N-butyl-3-butylpyridinium iodide has been synthesized. This molecule shows mixed reversible inhibition of acetylcholinesterase. The polymers also act as acetylcholinesterase inhibitors and show an unusual inhibition pattern. We tentatively describe it as quick initial reversible binding, followed by slow binding or irreversible inhibition of the enzyme. This kinetics suggests that there are several affinity binding sites on the acetylcholinesterase molecule where the polymer can bind. The first binding favors binding to other sites which leads to an apparently irreversibly linked enzyme-inhibitor complex. PMID- 9748588 TI - pH and volume homeostasis in trypanosomatids: current views and perspectives. PMID- 9748586 TI - Regulated expression of the homeobox gene, rPtx2, in the developing rat. AB - Using degenerate primers designed to amplify genes containing homeodomains, we have used reverse transcription and polymerase chain reaction to amplify and clone a rat homeobox gene. Based on the nucleotide and predicted amino acid sequences, the rat cDNA clone contains a high degree of sequence similarity to murine genes which are members of the paired-like class of homeobox genes (Ptx2, Otlx2, solurshin and Ptx1). Considering the high degree of sequence similarity and similar restricted expression patterns, we have named the cloned rat gene rPtx2 (rat Ptx2 homolog). Northern analysis revealed two rPtx2 transcripts expressed in the developing rat brain. Yet, only a single gene was detected by Southern blot hybridization, suggesting that multiple messages are the result of alternative transcriptional initiation, splicing or processing of a common message. The expression pattern of rPtx2 was further delineated by in situ hybridization to rat embryos. Within the brain, tissue specific expression was observed in the differentiating neural cells of the posterior hypothalamus, tegmentum, and rhombomere r1. Expression was also observed in the developing pituitary, maxilla, mandible, tongue and umbilical cord. To further study the control of Ptx2 gene expression, we used an in vitro model for neural differentiation by treating mouse embryonic stem cells with retinoic acid. Within 24 h and prior to detection of a neural phenotype in the culture, murine Ptx transcripts were induced and remained elevated for at least 6 days. This suggests that retinoic acid may be an important inductive signal which regulates the developmental and tissue-specific expression of Ptx2. PMID- 9748589 TI - The tachykinin NK1 receptor antagonist GR205171 prevents vagal stimulation induced retching but not neuronal transmission from emetic vagal afferents to solitary nucleus neurons in dogs. AB - Tachykinin NK1 receptor antagonists injected into the medulla oblongata are known to abolish vomiting induced by vagal afferent stimulation. Emetic vagal afferents have been shown to synapse with neurons in the medial solitary nucleus (mNTS), which suggests that substance P is a transmitter in the synapse. To examine this possibility, the effects of GR205171, an NK1 receptor antagonist, on retching and mNTS neuronal responses to the stimulation of abdominal vagal afferents were investigated in decerebrate dogs. GR205171 (0.05-0.7 mg kg-1, i.v.) abolished retching induced by either vagal or mNTS stimulation within 5 min. Firing of mNTS neurons in response to pulse-train and sustained vagal stimulation did not change even after the abolition of retching. Similarly, GR205171 did not have any effects on mNTS evoked potentials induced by pulse-train vagal stimulation. In about 20% of mNTS neurons, the peak firing frequency was facilitated to about 150% with repetitive pulse-train vagal stimulation. This facilitation remained even after the abolition of retching. Administration of GR205171 (1 mg ml-1, 30 microliters) into the 4th ventricle abolished retching, with latencies in excess of 120 min These results suggest that substance P does not participate in synaptic transmission between emetic vagal afferents and mNTS neurons in dogs. PMID- 9748590 TI - The mutagenic activity of ethylnitrosourea at low doses in spermatogonia of the mouse as assessed by the specific-locus test. AB - Ethylnitrosourea is the most efficient chemical mutagen in spermatogonial stem cells of the mouse and its mutagenic activity has been intensively studied. The pertinent specific-locus mutation test results for a discussion of low dose effect studies have been summarized and indicate: (1) A threshold dose response best characterizes the relationship between dose and mutation rate. (2) The reduced effectiveness of ethylnitrosourea in the low dose range is likely due to a saturable repair process. (3) The recovery of the saturable repair process as assessed in fractionated dose experiments is long (ca. 168 h). The dynamics of stem cell spermatogonia suggests a long time interval before the cell population passes through at least one cell division and this may be relevant to an interpretation of the fractionation effects. (4) There is a slight but important discrepancy between the predicted and observed mutagenic activity of ethylnitrosourea in the low dose range. This is interpreted to be due to the differences between a mathematical abstraction and the biological realities of the system being studied. PMID- 9748591 TI - The role of dietary choline in the beneficial effects of lecithin on the secretion of biliary lipids in rats. AB - Earlier studies showed that dietary soybean lecithin increases biliary lipid secretion, which mainly comes from the contribution of high density lipoprotein (HDL) and hepatic microsomal pools of phosphatidylcholine and cholesterol. In addition, a lecithin diet enhances bile secretion and prevents bile acid-induced cholestasis. This study evaluated the contribution of choline, a component of lecithin, to the observed effect of lecithin on biliary secretory function. Rats were fed either a control diet (CD), a choline diet (ChD) or a lecithin-enriched diet (LD) for 2 weeks. Results showed that like LD, ChD induced an increase in bile flow and bile acid secretion rate when compared with the control diet. However, unlike LD, ChD did not significantly increase biliary phospholipids and cholesterol output. An increase of hydrophilic bile acids (i.e. ursodeoxycholic and muricholic acids) in bile of rats fed choline could explain why the biliary phospholipid and cholesterol secretion was not increased. During taurocholic acid infusion, both experimental diets increased bile flow and the bile acid secretion rate maximum (BASRm). The cholestasis usually observed after the BASRm is reached was inhibited by ChD and LD. Both diets induced a decrease in plasma cholesterol (total and HDL), however, only LD induced statistically significant changes. Analysis of total cholesterol and phospholipid content of microsomes and canalicular membranes indicated no statistically significant difference between control and experimental groups either under basal conditions or after bile acid infusion. Similarly, the phospholipid classes and fatty acid composition of biliary phosphatidylcholine were not altered by feeding ChD and LD. We conclude that choline contributes to the beneficial effect of a lecithin diet on bile secretion. It is postulated that this effect may be attributed to modulation of HDL and an enhancement of the cholesterol and phospholipid pools destined for biliary secretion. PMID- 9748592 TI - The response of eukaryotic topoisomerases to DNA damage. AB - Beyond the known mutagenic properties of DNA lesions, recent evidence indicates that several forms of genomic damage dramatically influence the catalytic activities of DNA topoisomerases. Apurinic sites, apyrimidinic sites, base mismatches, and ultraviolet photoproducts all enhance topoisomerase I-mediated DNA cleavage when they are located in close proximity to the point of scission. Furthermore, when located between the points of scission of a topoisomerase II cleavage site, these same lesions (with the exception of ultraviolet photoproducts) greatly stimulate the cleavage activity of the type II enzyme. Thus, as found for anticancer drugs, lesions have the capacity to convert topoisomerases from essential cellular enzymes to potent DNA toxins. These findings raise exciting new questions regarding the mechanism of anticancer drugs, the physiological functions of topoisomerases, and the processing of DNA damage in the cell. PMID- 9748593 TI - Progressive internalization of beta-adrenoceptors in the rat liver during different phases of sepsis. AB - Changes in the distribution of beta-adrenoceptors (beta ARs) in the plasma membrane and the light vesicle fractions of rat liver during different phases of sepsis were studied using [3H]dihydroalprenolol binding and photoaffinity labeling with [125I]iodocyanopindolol diazirine. Sepsis was induced by cecal ligation and puncture (CLP). Septic rats exhibit an initial hypermetabolic (hyperglycemic) phase (9 h after CLP; early sepsis) followed by a hypometabolic (hypoglycemic) phase (18 h after CLP; late sepsis). The radioligand studies show that in the plasma membranes, the density of beta ARs was decreased by 28-32% and 46-69% during the early and the late phases, respectively, of sepsis. In the light vesicles, the density of beta ARs was increased by 25-30% and 30-35% during the early and the late phases, respectively, of sepsis. The total number of the receptor binding sites (the sum of that in plasma membrane plus light vesicle) was decreased by 11-12% and 21-35% during the early and the late phases, respectively, of sepsis. These results indicate that beta ARs were progressively internalized from surface membranes to the intracellular sites and, furthermore, they were underexpressed in the rat liver during the progression of sepsis. Since hepatic glucose metabolism is known to be regulated by catecholamines, in part, through beta AR mediation, an internalization/underexpression of hepatic beta ARs may play a role in the altered glucose homeostasis during sepsis, particularly in the late hypometabolic phase of sepsis. PMID- 9748595 TI - Postnatal development of dopamine D4-like receptors in rat forebrain regions: comparison with D2-like receptors. AB - Development of dopamine D4-like receptors in rat caudate-putamen (CPu), nucleus accumbens (NAc), frontal cortex, hippocampus, and entorhinal cortex was assessed at seven points between postnatal days 7 and 60 by computed in vitro receptor autoradiography, and compared with dopamine (DA) D2-like receptors. Density of radioligand binding to both receptor types increased from day 7 to a peak at day 28 in caudate-putamen (D4, 3.3-fold; D2, 4.3-fold) and nucleus accumbens (2.9- and 3.6-fold), then declined by 28%-33% over days 35-60 to adult levels in both brain regions. In hippocampus, frontal and entorhinal cortex, both receptor types increased by 3.8- to 5.8-fold from day 7 to maximal levels at day 35 that remained unchanged to day 60. These findings suggest: (1) D4- and D2-like receptors follow a similar course of development in several cortical, extrapyramidal, and limbic regions of rat forebrain; (2) elimination of excessive receptors of both types occurred in caudate-putamen and nucleus accumbens but not in the other brain regions. PMID- 9748594 TI - Oligomerisation of Ca2+-regulatory membrane components involved in the excitation contraction-relaxation cycle during postnatal development of rabbit skeletal muscle. AB - The skeletal muscle excitation-contraction-relaxation cycle matures during the first weeks after birth and protein-protein interactions are believed to be essential for proper Ca2+ regulation. We therefore studied potential changes in the oligomerisation of key components of the Ca2+-regulatory membrane system during postnatal myogenesis. In contrast to a decrease in calreticulin, the Ca2+ binding proteins calsequestrin and sarcalumenin increased in abundance in microsomes isolated from muscle between postnatal days 1 and 41. While the expression of the fast Ca2+-ATPase increased, its slow-twitch isoform decreased. The junctional component triadin, the 53 kDa sarcoplasmic reticulum glycoprotein, as well as the dihydropyridine receptor increased in abundance, while no major changes in the expression of the ryanodine receptor were observed. Crosslinking analysis revealed that the fast Ca2+-ATPase, alpha1-dihydropyridine receptor and calsequestrin exhibit a more pronounced tendency to oligomerise in adult muscle fibres as compared to early postnatal stages. Interestingly, adult calsequestrin exists not only as a 63 kDa form but also as stable molecular species of higher molecular mass. These findings imply that during postnatal development, protein protein interactions within the Ca2+-regulatory membrane system become more complex and oligomerisation appears to be an essential prerequisite for the proper physiological functioning of key membrane proteins in matured skeletal muscle fibres. PMID- 9748596 TI - Statistical models for low dose exposure. AB - Extrapolation of health risks from high to low doses has received a considerable amount of attention in carcinogenic risk assessment over decades. Fitting statistical dose-response models to experimental data collected at high doses and use of the fitted model for estimating effects at low doses lead to quite different risk predictions. Dissatisfaction with this procedure was formulated both by toxicologists who saw a deficit of biological knowledge in the models as well as by risk modelers who saw the need of mechanistically-based stochastic modeling. This contribution summarizes the present status of low dose modeling and the determination of the shape of dose-response curves. We will address the controversial issues of the appropriateness of threshold models, the estimation of no observed adverse effect levels (NOAEL), and their relevance for low dose modeling. We will distinguish between quantal dose-response models for tumor incidence and models of the more informative age/time dependent tumor incidence. The multistage model and the two-stage model of clonal expansion are considered as dose-response models accounting for biological mechanisms. Problems of the identifiability of mechanisms are addressed, the relation between administered dose and effective target dose is illustrated by examples, and the recently proposed Benchmark Dose concept for risk assessment is presented with its consequences for mechanistic modeling and statistical estimation. PMID- 9748597 TI - Decreased endothelial cell glutathione and increased sensitivity to oxidative stress in an in vitro blood-brain barrier model system. AB - Using a cell culture model of the blood-brain barrier (BBB) we have evaluated the role of endothelial cell glutathione in protecting barrier integrity against nitric oxide (NO)-induced oxidative stress. The co-culture of human umbilical vein endothelial cells (ECV304) with rat (C6) glioma cells, or incubation with glioma cell or primary astrocytic conditioned medium, resulted in a decline in endothelial cell glutathione. Exposure to a single addition of NO gas induced a rapid breakdown in model barrier integrity in endothelial/glioma co-cultures. Addition of NO gas or tumour necrosis factor-alpha (TNF-alpha) also resulted in a loss of membrane integrity, as measured by an enhanced release of lactate dehydrogenase, only from endothelial cells treated with glioma conditioned medium. Furthermore, assessment of viability in endothelial cells grown alone or treated with glioma conditioned medium, by propidium iodide labelled flow cytometry. demonstrated no difference in the number of positively stained cells after NO exposure. These results indicate that when enhanced endothelial monolayer barrier formation occurs via astrocytic-endothelial interactions, cellular glutathione levels are decreased. This renders the barrier cells, under these conditions, more susceptible to oxidative stress but does no necessarily lead to greater cell death. PMID- 9748598 TI - Secondary leukemias induced by topoisomerase-targeted drugs. AB - The major established cause of acute myeloid leukemia (AML) in the young is cancer chemotherapy. There are two forms of treatment-related AML (t-AML). Each form has a de novo counterpart. Alkylating agents cause t-AML characterized by antecedent myelodysplasia, a mean latency period of 5-7 years and complete or partial deletion of chromosome 5 or 7. The risk is related to cumulative alkylating agent dose. Germline NF-1 and p53 gene mutations and the GSTT1 null genotype may increase the risk. Epipodophyllotoxins and other DNA topoisomerase II inhibitors cause leukemias with translocations of the MLL gene at chromosome band 11q23 or, less often, t(8;21), t(3;21), inv(16), t(8;16), t(15;17) or t(9;22). The mean latency period is about 2 years. While most cases are of French American-British (FAB) M4 or FAB M5 morphology, other FAB AML subtypes, myelodysplastic syndrome (MDS), acute lymphoblastic leukemia (ALL) and chronic myelogenous leukemia (CML) occur. Between 2 and 12% of patients who receive epipodophyllotoxin have developed t-AML. There is no relationship with higher cumulative epipodophyllotoxin dose and genetic predisposition has not been identified, but weekly or twice-weekly schedules and preceding l-asparaginase administration may potentiate the risk. The translocation breakpoints in MLL are heterogeneously distributed within a breakpoint cluster region (bcr) and the MLL gene translocations involve one of many partner genes. DNA topoisomerase II cleavage assays demonstrate a correspondence between DNA topoisomerase II cleavage sites and the translocation breakpoints. DNA topoisomerase II catalyzes transient double-stranded DNA cleavage and rejoining. Epipodophyllotoxins form a complex with the DNA and DNA topoisomerase II, decrease DNA rejoining and cause chromosomal breakage. Furthermore, epipodophyllotoxin metabolism generates reactive oxygen species and hydroxyl radicals that could create abasic sites, potent position-specific enhancers of DNA topoisomerase II cleavage. One proposed mechanism for the translocations entails chromosomal breakage by DNA topoisomerase II and recombination of DNA free ends from different chromosomes through DNA repair. With few exceptions, treatment-related leukemias respond less well to either chemotherapy or bone marrow transplantation than their de novo counterparts, necessitating more innovative treatments, a better mechanistic understanding of the pathogenesis, and strategies for prevention. PMID- 9748599 TI - Renal and hepatic 1 alpha-hydroxylation of 25-hydroxyvitamin D3 in piglets suffering from pseudo vitamin D-deficiency rickets, type I. AB - The piglets examined suffer from rickets and have symptoms similar to those of classic pseudo vitamin D-deficiency rickets, type I (PVDRI), including plasma concentrations of 1 alpha, 25-dihydroxyvitamin D3 considerably lower than in healthy control piglets. It has been suggested that the rachitic piglets have a defective renal 1 alpha,25-dihydroxyvitamin D3 production. The present study shows that partially purified mitochondrial and microsomal cytochrome P450 from kidney and liver of both rachitic and control animals is able to catalyze 1 alpha hydroxylation of 25-hydroxyvitamin D3. The renal mitochondrial 1 alpha hydroxylase activity was higher in the rachitic piglets whereas the renal microsomal 1 alpha-hydroxylase activity was decreased. The immunodetectable levels in kidney of a mitochondrial 1 alpha-hydroxylase (CYP27) and a microsomal 1 alpha-hydroxylase (vitamin D3 25-hydroxylase) were correlated with the 1 alpha hydroxylase activities. The results suggest that the renal microsomal 1 alpha hydroxylase is affected by the rachitic condition. It is concluded that the primary genetic defect of systemic 1 alpha,25-dihydroxyvitamin D3 deficiency in the rachitic PVDRI piglets does not reside in a defective function or absence of renal mitochondrial 25-hydroxyvitamin D3 1 alpha-hydroxylase. From this, it may also be concluded that PVDRI in man and pig appear to be two different forms of the disease. PMID- 9748600 TI - Ontogenesis of hydroxyindole-O-methyltransferase gene expression and activity in the rat pineal gland. AB - Postnatal development of hydroxyindole-O-methyltransferase (HIOMT) mRNA expression, HIOMT activity and melatonin content was investigated in the rat pineal gland from birth to adulthood (62-day old). For each age, animals were sacrificed at two different time-points: midday and midnight. HIOMT mRNA was first detectable one day after birth and maximal diurnal levels were reached at the end of the first week. A 2-fold nocturnal increase appeared significantly 11 days after birth. HIOMT activity was detectable from 5 days of life and significant day/night variations did not appear before 21 days after birth. Appearance of melatonin synthesis and rhythm followed that of HIOMT mRNA. The 10 day delay observed between the appearance of the nocturnal increase in HIOMT activity and expression is discussed in term of differential regulation of both HIOMT mRNA expression and HIOMT activity. PMID- 9748601 TI - An evaluation of the marmoset Callithrix jacchus (sagui) as an experimental model for the dyslipoproteinemia of human Schistosomiasis mansoni. AB - Human infection with the parasite Schistosoma mansoni is a relatively common occurrence in regions of South America and is associated with liver dysfunction and dyslipoproteinemia. Specifically, the activity of plasma lecithin:cholesterol acyltransferase (LCAT) activity is reduced, the concentration of plasma cholesterol esters falls, phospholipid concentrations are elevated and erythrocyte membranes become cholesterol enriched. Previous studies have utilized rodents (rats and mice) as experimental models to study the dyslipoproteinemia induced by S. mansoni infection. However, the plasma lipoprotein profiles in these animals is very different from humans and infection is not accompanied by decreases in LCAT activity or cholesterol enrichment of their erythrocyte membranes. Here we have evaluated the suitability of the marmoset Callithrix jacchus (sagui) which is small and readily available in Brazil, as a potential animal model for the study of the dyslipoproteinemia of S. mansoni infections. The plasma lipoprotein compositions and distributions in sagui, unlike rats or mice, approximate those of man with the LDL representing a major lipoprotein species. The molecular species of phospholipids, cholesterol esters and triglycerides present in sagui plasma are also very similar to man, whereas those of rats and mice favor the longer chain more unsaturated species, Sagui, like rodents, can be successfully infected with S. mansoni and after 60 days, this results in a 50% reduction in plasma LCAT activity, an 11% reduction in plasma cholesterol esters, an absolute increase of 46% in plasma phospholipids and an 18% increase in the cholesterol content of erythrocyte membranes. These changes are qualitatively and quantitatively very similar to those previously reported following human infections. Based upon these changes, and the observation that the plasma lipoprotein profile of sagui and human is similar, we conclude that C. jacchus (sagui) is an appropriate animal model for the study of dyslipoproteinemia associated with S. mansoni infections. PMID- 9748602 TI - Population monitoring: experience with residents exposed to uranium mining/milling waste. AB - More emphasis should be placed upon using biomarkers to address potential health risk among populations exposed to high concentrations of environmental toxicants. Among these studies, those which integrate exposure measurements with analyses of validated biomarkers may provide more reliable information for risk assessment and disease prevention. We have used a multidisciplinary approach to elucidate potential health hazards in a population living around uranium mining/milling facilities. The study included 24 target and 24 control residents who were matched for age and gender and selected based on time of residence in the study areas and proximity to mining/milling sites. Environmental samples were analyzed for uranium-238 (238U) concentrations and lead isotope ratios using inductively coupled plasma-mass spectrometry (ICP-MS) procedures, and blood samples were collected for cytogenetic analysis. We found that the 238U concentrations in soil samples were significantly higher than those in the control areas. In addition, the concentrations in the surface soil were significantly higher than in the subsurface soil (p<0.05) from target areas indicating environmental contamination by the mining/milling activities. Lead isotope data from soil samples taken near a railroad transfer location was significantly different from those of other sites, indicating contamination by non-native ore transported from sources outside of the region to local milling facilities for processing. Therefore, local residents have been exposed to low levels of radioactive contamination from the mining/milling activities on a daily basis for many years. From our cytogenetic analysis, the target population had more chromosome aberrations than the controls, although the differences were not significant (p<0.05). However, using our challenge assay, cells from the target population had a significantly abnormal DNA repair response, compared to cells from the same control population. In conclusion, the observed environmental contamination by uranium is consistent with the observed genotoxic effects in the target residents. Therefore, the residents have increased health risk and some of the health problems will most likely be related to exposure to the radioactive contaminants. Since the chromosome aberration frequency revealed increased, but not significant differences between the exposed and the control populations, we conclude that the health risk among the exposed residents is similar to those among nuclear workers. PMID- 9748603 TI - Structure-activity analysis of brevinin 1E amide, an antimicrobial peptide from Rana esculenta. AB - Brevinin 1E, consisting of 24 amino acid residues, from Rana esculenta has potent antimicrobial and hemolytic activity. From a structural point of view, this peptide has a N-terminal hydrophobic region, a proline hinge region in the middle and a C-terminal loop region delineated by an intra-disulfide bridge, which is a common structural feature of antimicrobial peptides from Rana species. To investigate the structural features for antimicrobial and hemolytic activity, truncated and linearized brevinin 1E amides were synthesized and characterized. A deletion of three amino acids from the N-terminal region did not greatly affect antimicrobial activity but dramatically reduced hemolytic activity. The contribution of the intra-disulfide bridge to antimicrobial and hemolytic activity was somewhat different between brevinin 1E amide and truncated fragments. In brevinin 1E amide, the elimination of the intra-disulfide bridge did not greatly affect antimicrobial and hemolytic activity whereas the elimination of the intra-disulfide bridge in the truncated fragments did not decrease antimicrobial activity but did decrease hemolytic activity. Circular dichroism spectra and the retention time on the C18 reverse phase column revealed that the intra-disulfide bridge (i, i+6) formed an amphipathic loop which increased hydrophobicity and helped to induce the alpha-helical structure in the membrane-mimetic environment. Even though the intra-disulfide bridge and the N terminal region were responsible for the alpha-helical structure and hydrophobicity, these two structural features were not essential for antimicrobial activity. The hemolytic activity of brevinin 1E amide and its analogs also correlated well with the retention time rather than the alpha helicity. PMID- 9748604 TI - Decreased central histamine in the amygdaloid kindling rats. AB - This study was conducted to elucidate the role of central histamine (HA) in seizure susceptibility. We stimulated the left amygdala of rats to produce amygdaloid kindling. We sacrificed rats 1 h, 1 week and 1 month after the last kindled seizure, and measured the histamine contents and the histidine decarboxylase (HDC) activities of various brain regions. One hour after the last kindled seizure, we found significant decreases in HA levels in the bilateral amygdala, hippocampus and diencephalon in the kindled group. The HDC activities of the bilateral amygdala and diencephalon were lower in the kindled group than in the control group. One week after the last kindled seizure, we also found a significant decrease in the HA level in the bilateral amygdala. No significant change was found in HA content or HDC activity 1 month after the last kindled seizure. These results suggest that kindling suppresses HA synthesis and that the reduced HA content is maintained until 1 week after the last kindled seizure. The reduced HA may play a role in the acquired kindled seizure susceptibility. PMID- 9748605 TI - Experience-dependent changes in the importance of N-methyl-D-aspartate (NMDA) receptors for visual transmission in superior colliculus. AB - The excitatory amino acid transmitter glutamate mediates visual activity in the superficial grey layer (SGS) of superior colliculus. At eye opening N-methyl-d aspartate receptors (NMDA-rs) convey little of the visual response, but with age their role in visual transmission increases to a peak at P21, then falls to the lower adult level. Visual deprivation which begins before eye opening causes NMDA rs to assume a greater importance for visual transmission in SGS. Here we explore the possibility that these experience-dependent changes in the role of NMDA-rs in the SGS are limited by age. We find that the effects of visual deprivation on NMDA-r mediated visual activity are recoverable even after extensive dark rearing. Also, a short episode of visual experience is sufficient to allow the normal situation to be established and subsequent dark rearing is ineffective. Four-day periods of visual experience beginning at P14 or P25 have the same effect. Given that NMDA-rs take little part in visual transmission prior to P18, these data prompt a reconsideration of the role of NMDA-r mediated sensory transmission in the mechanisms by which early environmental experience influences the development of the visual system. PMID- 9748606 TI - Hypothyroidism renders liver mitochondria resistant to the opening of membrane permeability transition pore. AB - Membrane permeability was examined in liver mitochondria isolated from hypothyroid rats. It was found that such a thyroid status provides substantial protection from membrane leakiness as induced by Ca2+ loading. Thus, these mitochondria are less prone to undergoing permeability transition than mitochondria from euthyroid rats. The above conclusion was reached on the basis of the following two facts: (1) hypothyroid mitochondria are not strictly dependent on the addition of ADP to retain high matrix Ca2+ concentrations, and (2) carboxyatractyloside, antimycin A or carbonyl cyanide-m-chlorophenyl hydrazone failed to promote Ca2+ efflux. We discuss the possible relevance of the low content of membrane cardiolipin as well as the low expression of the adenine nucleotide translocase as responsible for the resistance to membrane damage. PMID- 9748607 TI - IL-13 increases the cPLA2 gene and protein expression and the mobilization of arachidonic acid during an inflammatory process in mouse peritoneal macrophages. AB - Pretreatment of mouse peritoneal macrophages with interleukin-13 (IL-13) potentiates the mobilization of arachidonic acid (AA) and the production of HETEs but does not affect the production of cyclooxygenase metabolites triggered by the suboptimal concentration of an inflammatory agonist (opsonized-zymosan). Cycloheximide suppresses these effects of IL-13 suggesting that de novo protein synthesis is involved. Indeed, IL-13 induces a time-dependent increase in the levels of cytosolic PLA2 (cPLA2) protein and mRNA. This study demonstrates a new pathway for IL-13 to modulate the inflammatory process in macrophages via modifications of cPLA2 expression and subsequent AA mobilization. PMID- 9748608 TI - Stimulation of nerve growth-factor and substance P expression in the iris trigeminal axis of diabetic rats--involvement of oxidative stress and effects of aldose reductase inhibition. AB - In rats with streptozotocin-induced diabetes, we measured increased (by 61%; P < 0.05) mRNA for nerve growth factor (NGF) in the iris together with increased (by 82%; P < 0.05) mRNA for preprotachykinin (the substance P precursor) in the trigeminal ganglion, suggesting that increased NGF was driving increased substance P gene expression. In other diabetic rats, these changes were prevented by treatment with either an antioxidant (butylated hydroxytoluene; 1% by diet) or an aldose reductase inhibitor (ARI) (sorbinil; 25 mg/kg/day p.o.) and the sorbinil treatment was associated with significant inhibition of polyol pathway intermediates in both lens and sciatic nerve. This suggests that polyol pathway activity in the lens may translate to oxidative stress-driving stimulation of NGF gene expression in the iris. The change is selective for NFG, because expression of the analogous neurotrophin, neurotrophin-3 (NT-3), was unaltered in the same irises. These changes suggest that oxidative stress and/or inflammation can drive up NGF expression in diabetes--a mechanism that might participate in iritis. PMID- 9748609 TI - Butadiene: species comparison for metabolism and genetic toxicology. AB - The synthetic monomer 1,3-butadiene and its metabolites have been reviewed in various in vitro and in vivo metabolic studies and in genetic toxicology assays. The species differences have been compared. PMID- 9748610 TI - Genotype-phenotype correlations in mucopolysaccharidosis type I using enzyme kinetics, immunoquantification and in vitro turnover studies. AB - Fibroblasts from 16 patients with known alpha-L-iduronidase gene mutations and different clinical phenotypes of mucopolysaccharidosis type I (MPS I) were investigated in order to establish genotype/phenotype correlations. Enzyme kinetic studies were performed using the specific alpha-L-iduronidase substrate iduronosyl anhydro[1-3H]mannitol-6-sulfate. Specific residual enzyme activities were estimated using the kinetic parameters and an immunoquantification assay which determines levels of alpha-L-iduronidase protein. Cells were cultured in the presence of [35S]sulfate and the in vivo degradation of accumulated labelled glycosaminoglycans measured after different chase times. Residual enzyme activity and different amounts of residual enzyme protein were present in extracts from 9 of 16 cell lines covering a wide spectrum of clinical severity. Catalytic capacity, calculated as the product of kcat/Km and ng iduronidase protein per mg cell protein, was shown in most cases to be directly related to the severity of clinical phenotype, with up to 7% of normal values for patients with the attenuated form of MPS I (Scheie) and less than 0.13% for severely affected patients (Hurler) In vitro turnover studies allowed further refinement of correlations between genotype and phenotype. Scheie disease compared to Hurler disease patients were shown to accumulate smaller amounts of glycosaminoglycans that were also turned over faster. A combination of turnover and residual enzyme data established a correlation between the genotype, the biochemical phenotype and the clinical course of this lysosomal storage disorder. PMID- 9748611 TI - Thiol-dependent metal-catalyzed oxidation of copper, zinc superoxide dismutase. AB - Superoxide dismutase (SOD) is a key enzyme in the antioxidant system of the cells. When exposed to a metal-catalyzed oxidation (MCO) system composed of Fe3+, O2, and thiol as an electron donor copper, zinc SOD (CuZnSOD) was susceptible to oxidative modification and damage as indicated by the loss of activity, fragmentation and aggregation of peptide as well as by the formation of carbonyl groups. Oxidative damage to CuZnSOD was inhibited by diethylenetriaminepentaacetic acid as well as by free radical scavengers and spin trapping agents. The results of the present study indicate that hydrogen peroxide may be generated from a thiol/Fe3+/O2 system and that hydroxyl free radicals, produced by metal-catalyzed Fenton reactions, may be the ultimate species mediating the SOD damage. Incubation with the MCO system resulted in the release of Cu ions from CuZnSOD. Incubation with the thiol-MCO did not significantly increase the formation of 2-oxohistidine in CuZnSOD. The lack of formation of 2 oxohistidine, as well as the pronounced preventive effect of spin-traps on the thiol-MCO-mediated damage to CuZnSOD, indicates that inactivation might actually be predominantly due to global oxidation rather than a site-specific oxidation. The thiol-MCO-mediated damage to SOD may result in the perturbation of cellular antioxidant defense mechanisms and subsequently lead to a pro-oxidant condition. PMID- 9748612 TI - Prosaposin prevents programmed cell death of rat cerebellar granule neurons in culture. AB - Prosaposin, the precursor of sphingolipid activator proteins (saposin A-D), has been reported to be a neurotrophic factor in vitro and in vivo. Prosaposin mRNA is transiently expressed at a high level in developing cerebellum during the period of granule cell proliferation and maturation, suggesting its significance during development of cerebellum. Here we investigated the neuroprotective effect of prosaposin on cerebellar granule neurons, exposing primary cerebellar granule cells to low K+ which induced programmed cell death. Prosaposin rescued mature cerebellar granule neurons in a bimodal manner. A similar neuroprotective effect was obtained using TX14(A), a 14-mer neurotrophic peptide derivative of prosaposin. An additive neuroprotective effect was observed between BDNF and TX14(A), but not between IGF-1 and TX14(A). Prosaposin rescued 60% of nifedipine sensitive cerebellar granule neurons as well as IGF-1, while BDNF did not. Furthermore, the neuroprotective action of prosaposin was inhibited by LY294002, a specific inhibitor of PI 3-kinase. These findings indicated that prosaposin had a trophic effect upon newborn cerebellar granule cells and that the neuroprotective action was similar to that of IGF-1 rather than BDNF. Prosaposin may play a role in cerebellar development during programmed cell death of cerebellar neurons. PMID- 9748613 TI - Mechanisms that account for the selective release of arachidonic acid from intact cells by secretory phospholipase A2. AB - The current study examined mechanisms that account for the selective release of arachidonic acid (AA) from cells by secretory phospholipase A2 (sPLA2). Initial studies demonstrated that low concentrations of group I and group III PLA2 isotypes and an sPLA2-enriched extract from bone marrow-derived mast cells (BMMC) selectively released AA from mast cells. Much higher concentrations of group II PLA2 were required to release comparable quantities of AA. Group I PLA2 also selectively released AA from another mast cell line (CFTL-15) and a monocytic cell line (THP-1). In contrast, high concentrations of group I PLA2 were required to release fatty acids from a promyelocytic cell line (HL-60) and this release was not selective for AA. Binding studies revealed that cell types (BMMC, CFTL-15 and THP-1) which selectively released AA also had the capacity to specifically bind group I PLA2. However, group II PLA2, which did not selectively release AA from cells, also did not specifically bind to these same cell types. Additional studies revealed that sPLA2 binding to the mast cell receptor was attenuated after stimulation with antigen or ionophore A23187. Reverse transcriptase polymerase chain reaction analyses indicated the presence of mRNA for the sPLA2 receptor in BMMC, CFTL-15 and THP-1 and the absence of this mRNA in HL-60. Final studies demonstrated that p-aminophenyl-alpha-D-mannopyranoside BSA, a known ligand of the sPLA2 receptor, also selectively released AA from mast cells but not from HL-60 cells. These experiments indicated that receptor occupancy alone (without PLA2 activity) is sufficient to induce the release of AA from mast cells. Together, these data reveal that specific isotypes of sPLA2 have the capacity to selectively release AA from certain cells by their capacity to bind to sPLA2 receptors on the cell surface. PMID- 9748614 TI - Effect of macrophage suppression with silica on the proliferation of Schwann cells during Wallerian degeneration. AB - Effects of macrophage suppression on Schwann cell proliferation in Wallerian degeneration was investigated in vitro with thymidine autoradiography. Sciatic nerves of C57BL mice were transected shortly after an intraperitoneal injection of silica dust and Schwann cells were harvested at 1, 2, 3, 5, 7, and 10 days post-transection. In silica treated mice, Schwann cells from post-transected nerve stumps proliferated despite marked suppression of macrophages, although there was some initial delay. The results of this study suggest that the mechanism of Schwann cell proliferation is multifactorial and in addition to the influence of growth factors mediated and released by macrophages, other factor(s) such as degenerated axolemma may play a more pivotal role in Schwann cell proliferation in vivo. PMID- 9748615 TI - Inhibition of long-term potentiation in developing rat visual cortex but not hippocampus by in utero exposure to polychlorinated biphenyls. AB - The neurotoxic potential of polychlorinated biphenyls (PCBs) depends on the structure of the congener as well as on the age of the exposure. We exposed rats prenatally to a coplanar congener (PCB-77) or to a non-coplanar congener (PCB-47) and measured the amount of long-term potentiation (LTP) at postnatal days 11-19 in the visual cortex and hippocampus. While PCB-77 exposure affected LTP statistically significantly in cortical but not hippocampal slices, the exposure to PCB-47 was much less effective. PMID- 9748616 TI - Isoform-specific effects of salts on nitric oxide synthase activity. AB - We investigated the effects of salts on the properties of the neuronal, endothelial, and inducible isoforms of nitric oxide synthase (nNOS, eNOS, and iNOS), and found pronounced isoform-specific effects on NOS-catalyzed L citrulline formation. Salts inhibited iNOS monotonously, whereas nNOS and eNOS were stimulated up to 3-fold at low, and inhibited at high (>/=0.1-0.2 M) salt concentrations. The effectivities of different ions mostly followed the Hofmeister series, indicating that the effects can for a large part be ascribed to changes in protein solvation. Km(Arg) increased in the presence of NaCl, demonstrating the importance of charge interactions for substrate binding. The coupling of NADPH oxidation to NO production was not affected by KCl. Salts (Glu)). Wild-type (wt) PAI 1 revealed specific activities of 80+/-9% (mean+/-S.D., n=4) of the theoretical maximum value towards t-PA. PAI-1-ovalbumin, PAI-1-antithrombin III and PAI-1-P13 (Val-->Glu) revealed specific activities of 86+/-15%, 77+/-11%, and 100+/-30% respectively, towards t-PA and similar inhibitory properties towards u-PA. Surprisingly, upon inactivation at 37 degreesC, the active conformation of the PAI-1 mutants converted partly into a substrate conformation (i.e. 52+/-5.2%, 55+/-8.2% and 46+/-4.6% for PAI-1-ovalbumin, PAI-1-antithrombin III and PAI-1-P13 (Val-->Glu), respectively) and partly into a latent conformation. This is in contrast to active wtPAI-1 which, as expected, is converted to the latent conformation (i.e. 86+/-1.0%). In conclusion, even though replacement of the P13 to P10 region of PAI-1 by the corresponding region of a non-inhibitory serpin or of an inhibitory serpin, does not directly affect its inhibitory properties, the nature of the amino acids in this region and of P13 in particular, contributes to its conformational transitions. PMID- 9748635 TI - Manganese-stimulated phosphatidylinositol headgroup exchange in rat liver microsomes. AB - Manganese-dependent, CMP-independent incorporation of myo-[3H]inositol into phospholipids of rat liver microsomes was studied in an attempt to clarify the physiological significance of this headgroup-exchange reaction. The enzyme responsible worked best with Mn2+ as a co-factor, but Mg2+ at physiological concentrations supported a significant rate of incorporation. The K(m) for myo inositol was around 11 microM, yet incorporation of myo-[3H]inositol was unaffected by as much as 5 mM choline, ethanolamine, glycerol or serine; as this is a reversible reaction, these data imply that phosphatidylinositol is the most likely lipid substrate. Similarly, other inositols showed an apparent affinity at least two orders of magnitude lower than myo-inositol. Glucosamine alpha 1-6 myo inositol also had a low affinity for the enzyme, making it unlikely that this headgroup-exchange activity is part of a metabolic pathway for glycosyl phosphatidylinositols. The phosphatidylinositol radiolabelled by headgroup exchange was deacylated and deglycerated, and the resulting inositol phosphate headgroup cochromatographed on anion exchange HPLC with myo-inositol l-phosphate. The simplest interpretation of all the data is the apparent paradox that this enzyme functions at a slow rate under physiological conditions to remove the myo inositol headgroup from phosphatidylinositol, only to replace it with another myo inositol. PMID- 9748636 TI - Distributions of periventricular projections of the paraventricular nucleus to the median eminence and arcuate nucleus. AB - Neuronal projections from the periventricular subnucleus of the hypothalamic paraventricular nucleus to the median eminence and the arcuate nucleus were investigated in the rat by the anterograde tract-tracer, Phaseolus vulgaris leucoagglutinin. The vast majority of labeled fibers coursed ventrally along the third ventricle and distributed in the external layer of the median eminence bilaterally, with ipsilateral predominance moving caudalwards. Periventricular fibers also terminated in the arcuate nucleus, but this innervation was exclusively ipsilateral. PMID- 9748637 TI - Organization and structure of NADH-dependent glutamate synthase gene from rice plants. AB - Genomic clones for NADH-dependent glutamate synthase (NADH-GOGAT; EC 1.4.1.14) were obtained from a genomic library of rice (Oryza sativa L. cv. Sasanishki). A genomic clone (lambdaOS42, 14 kb) covered an entire structural gene and a 3.7 kb 5'-upstream region from the first methionine. Another clone (lambdaOS23, 14 kb) contained a 2.8 kb 3'-downstream region from the stop codon. A 7047 bp long clone (lambdaOSR51) consisting of full length cDNA for NADH-GOGAT was isolated from a cDNA library prepared using mRNA from roots of rice seedlings treated with 1 mM NH4Cl for 12 h. The presumed transcribed region (11.7 kb) consisted of 23 exons separated by 22 introns. Rice NADH-GOGAT is synthesized as a 2166 amino acid protein with a molecular mass of 236.7 kDa that includes a 99 amino acid presequence. DNA gel blot analysis suggested that NADH-GOGAT occurred as a single gene in rice. Primer extension experiments map the transcription start of NADH GOGAT to identical positions. The 3. 7 kb 5'-upstream region was able to transiently express a reporter gene in cultured rice cells. Putative motifs related to the regulation of NADH-GOGAT gene expression were looked for within the 5'-upstream region by database. PMID- 9748639 TI - Animal models for studying the action of topoisomerase I targeted drugs. AB - Almost 30 years after the unsuccessful clinical evaluation of camptothecin sodium, there has been a revival in interest in this class of agent that poisons topoisomerase I. Currently there are four camptothecin analogues in clinical trials each at different levels of advancement. Clinical data suggest that patterns of antitumor activity and toxicity profiles differ between analogues. In preclinical models antitumor activity appears to be highly schedule-dependent. Here we review rodent and human tumor models used in evaluation of efficacy, and models used to predict toxicities of these compounds. The major limitation of rodent models is that the mouse tolerates significantly greater systemic exposure to each camptothecin analogue than do patients. This leads to a false overprediction of potential clinical activity. However, responses of human tumor xenografts in mice are highly predictive of responses of clinical cancer when camptothecins are administered at dose levels achieving similar systemic exposure in mice. Development of assays that identify analogues that maintain therapeutic activity in mice, but have less species differential toxicity, particularly to the hematopoietic system, may provide an early screen to select compounds having greater clinical utility. PMID- 9748638 TI - Methylene-interrupted double bond in polyunsaturated fatty acid is an essential structure for metabolism by the fatty acid chain elongation system of rat liver. AB - Some plant oils contain non-methylene-interrupted polyunsaturated fatty acids (NMIFAs). Pinolenic acid (all cis delta-5,9,12/18:3) and columbinic acid (trans,cis,cis delta-5,9,12/18:3) are NMIFAs that exist in pine seed oil and columbine seed oil, respectively. We investigated the double bond position of fatty acid recognized by the fatty acid chain elongation system (FACES) of rat liver using NMIFAs as experimental tools. In the total elongation assay, amounts of C2 unit chain-elongated metabolites of pinolenic acid and columbinic acid were 32% and 11%, respectively, compared to that of gamma-linolenic (all cis delta 6,9,12/18:3) as the substrate. In the condensation reaction assay, the rate limiting step of FACES, the conversion rates of pinolenic acid and columbinic acid to the corresponding C20 beta-keto fatty acids were 19% and 9% of that of gamma-linolenic acid, respectively. The formation of elongated metabolite of podocarpic acid (all cis delta-5,11,14/20:3) was only 7% of that of arachidonic acid (all cis delta-5,8,11,14/20:4). From these results it was concluded that the condensing enzyme of FACES could recognize the methylene-interrupted cis double bond structure vicinal to the carboxyl group in the fatty acid molecule. PMID- 9748640 TI - An intracellular role for pancreatic bile salt-dependent lipase: evidence for modification of lipid turnover in transfected CHO cells. AB - Pancreatic bile salt-dependent lipase (BSDL) hydrolyzes cholesteryl esters, triglycerides and phospholipids. BSDL is also capable of transferring free fatty acid to cholesterol. BSDL has been detected in many cells including fetal and tumor cells, hepatocytes, macrophages and eosinophils and in tissues such as adrenal glands and testes. The enzyme may be secreted or located within subcellular compartments such as the endoplasmic reticulum or the cytosol. Although the role of the secreted enzyme is well documented, that of the intracellular form(s) is still hypothetical. In the present study, we addressed the effects of BSDL on cell lipid metabolism. For that purpose, the cDNA of rat BSDL was transfected into CHO K1 cells (CHO K1-BSDL clone) which were then loaded with [3H]oleic acid. The results demonstrate that the transfected BSDL is secreted; in spite of that, a large fraction of catalytically active BSDL is found in cell lysate. The lipid metabolism of transfected cells is affected and BSDL induces an enhanced incorporation of [3H]oleic acid in cholesteryl esters whereas fatty acid incorporation in phosphatidylcholine is decreased. These effects were particularly important in the cytosol of transfected cells where transfected BSDL preferentially locates. These data suggested that BSDL could be implicated in the cycle of the cellular homeostasis of cholesterol which is particularly affected in tumoral cells leading to cholesteryl ester storage within cytosolic lipid droplets. PMID- 9748641 TI - Specificity of a wheat gluten aspartic proteinase. AB - The substrate and peptide bond specificities of a purified wheat gluten aspartic proteinase (GlAP) are studied. GlAP shows maximum gluten hydrolysing activity at pH 3.0. At this pH, especially the wheat high molecular weight glutenin subunits (HMW-GS) and to a lesser extent the low molecular weight glutenin subunits and gliadins are hydrolysed. GlAP has no obvious effect on albumins and globulins. In its action on oxidised insulin B-chain, GlAP forms eight peptides and has high specificity for peptide bonds located between amino acid residues with large hydrophobic side chains (Leu, Phe, Tyr) but the peptide bond Glu13-Ala14 is also hydrolysed. Although structurally quite similar to a barley aspartic proteinase, the peptide bond specificity of GlAP towards oxidised insulin B-chain resembles slightly more that of a cardoon aspartic proteinase, cardosin B. HMW-GS 7, purified from cultivar Galahad-77, is rapidly hydrolysed by GlAP. N-Terminal amino acid sequence data show that GlAP cleaves at least one Met-Ile peptide bond at the end of the N-terminal domain and two Val-Leu peptide bonds in the repetitive domain of HMW-GS 7. PMID- 9748642 TI - Homocysteine stimulates the production and secretion of cholesterol in hepatic cells. AB - Homocysteinemia and hypercholesterolemia are important risk factors associated with the occurrence of arteriosclerotic vascular diseases. A positive correlation between plasma levels of homocysteine and cholesterol was found in homocysteinemic patients as well as in experimental animals. In the present study, the effect of homocysteine on the production and secretion of cholesterol in human hepatoma cell line HepG2 cells was investigated. When cells were incubated with 4 mM homocysteine, the amounts of total cholesterol produced as well as the cholesterol secreted by these cells were significantly increased (from 32 +/- 5 to 74 +/- 5 nmol/mg cellular protein). Further biochemical analyses revealed that the increase in cholesterol was resulted from an enhancement in the production and secretion of the unesterified cholesterol with no concomitant change in the level of cholesteryl esters. The activity of intracellular 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase was markedly elevated by 131% and 190% after cells were incubated with homocysteine for 24 and 48 h. Homocysteine also stimulated the secretion of apo B100 by HepG2 cells (from 0.84 +/- 0.11 to 1.37 +/- 0.12 micrograms apolipoprotein B/mg cellular protein). Our results demonstrate that homocysteine stimulates the production and secretion of cholesterol and apolipoprotein B100 in HepG2 cells. The increase in the production of cholesterol induced by homocysteine may contribute to the pathogenesis of arteriosclerosis. PMID- 9748643 TI - Vaccinia virus DNA topoisomerase: a model eukaryotic type IB enzyme. AB - Vaccinia topoisomerase has proven to be an instructive model system for mechanistic studies of the type IB family of DNA topoisomerases. The catalytically relevant functional groups at the active site and the circumferential topoisomerase-DNA interface were correctly surmised by mutational and footprint analysis of vaccinia topoisomerase in advance of structure determinations by X-ray crystallography. It is now evident from multiple crystal structures that the catalytic domains of type IB topoisomerases and site specific recombinases derive from a common ancestral strand transferase capable of forming a DNA-(3'-phosphotyrosyl)-enzyme intermediate. A constellation of conserved amino acids catalyzes attack of the tyrosine nucleophile on the scissile phosphate. Domain dynamics and DNA-induced conformational changes within the catalytic domain are likely to play a role in triggering strand scission and coordinating the strand exchange or strand passage steps. PMID- 9748645 TI - Enhancement of phospholipase D activity following baculovirus and adenovirus infection in Sf9 and COS-7 cells. AB - In order to purify the human phospholipase D1 (hPLD1) for analysis of its functional properties, we applied a baculovirus-based high-expression system. As expected, Sf9 cells infected with a baculovirus encoding for the hPLD1 displayed a 7.5-fold increase in PLD activity compared to uninfected cells. Sf9 cells infected with the wild-type (WT) and other recombinant baculoviruses were used as an expression control. Surprisingly, all baculoviruses tested led to a 3-5 fold increase in basal PLD activity when compared to uninfected cells. To further characterize the nature of the increased PLD activity, the influence of ADP ribosylation factor (ARF) and phorbol 12-myristate 13-acetate (PMA) was studied. In contrast to membranes containing the hPLD1, the PLD activity in membranes from uninfected and WT-infected Sf9 cells was not stimulated by ARF. PMA did not affect the increase in PLD activity in any case. To further study whether the virus-mediated increase in PLD activity is a more general phenomenon, we infected COS-7 cells with recombinant and WT adenoviruses. Only the infection with the WT adenovirus resulted in an approx. 2-fold increase in PLD activity. Our results demonstrate for the first time that a viral infection elevates the PLD activity in insect and mammalian cells. PMID- 9748644 TI - Potential use of a novel lipase from Aspergillus carneus in deacetylation reactions. AB - A novel lipase from Aspergillus carneus has been used in organic solvents for efficient regioselective and chemoselective deacetylation of the peracetates of polyphenolic aromatic ketones, esters and amides. A reversal of regioselectivity was observed as compared with the results obtained during deacetylation with porcine pancreatic lipase (PPL). PMID- 9748646 TI - Pancreatic lipase-related protein 1 (PLRP1) is present in the pancreatic juice of several species. AB - Pancreatic lipase-related protein 1 (PLRP1) was purified from human, canine, porcine and rat pancreatic juices. The four PLRP1s were identified using microsequencing methods after performing gel filtration on Ultrogel AcA-54 followed by chromatography on Heparin-Sepharose cation-exchanger. Polyclonal antibodies specific to human PLRP1 (HPLRP1) were raised in the rabbit using a synthetic decapeptide from HPLRP1. The results of Western blotting analysis showed that these antibodies recognized native HPLRP1 and recombinant HPLRP1 produced by insect cells, and cross-reacted only with rat PLRP1 (RPLRP1). No significant lipolytic activity was observed with native canine PLRP1 and recombinant HPLRP1 on various glycerides, phospholipid and vitamin esters, or on cholesterol esters. It was established for the first time that this protein is secreted in variable amounts by the adult exocrine pancreas of several species. PMID- 9748647 TI - Catabolic fate of dietary trilinoleoylglycerol hydroperoxides in rat gastrointestines. AB - To elucidate whether dietary lipid peroxides are absorbed in the body, the catabolic fate of trilinoleoylglycerol hydroperoxides (TL-OOH), in the gastrointestines of rats was examined. Oxidized trilinoleoylglycerol with a peroxide value of 1000 meq/kg, 0.5 or 20 mg, was dosed intragastrically to rat together with 59.5 or 40 mg unoxidized trilinoleoylglycerol, respectively. The fate of TL-OOH in gastric and intestinal lumina was determined by high performance liquid chromatography periodically until 240 min after treatment. At low dose, TL-OOH was soon broken down to linoleic acid hydroperoxides (LA-OOH) and hydroxyls, probably through gastric lipases, whereas at high dose, TL-OOH was retained in the stomach. In both cases, TL-OOH did not reach the intestines, though the unoxidized lipids moved to the intestines. When LA-OOH was given intragastrically, the lipids decomposed in the stomach, and linoleic acid hydroxyls, hexanal, 9-oxononanoic acid, and two novel compounds were detected 30 min after treatment. The novel compounds were identified to be epoxyketones, 11 oxo-12,13-epoxy-9- and 11-oxo-9,10-epoxy-12-octadecenoic acids. Thus, dietary TL OOH was broken down in the stomach releasing, LA-OOH which decomposed further, and did not reach the intestines. PMID- 9748648 TI - Bacteriophage T4, a model system for understanding the mechanism of type II topoisomerase inhibitors. AB - Bacteriophage T4 provides a simple model system for analyzing the mechanism of action of antitumor agents that inhibit DNA topoisomerases. The phage-encoded type II topoisomerase is sensitive to many of the same antitumor agents that inhibit mammalian type II topoisomerase, including m-AMSA, ellipticines, mitoxantrone and epipodophyllotoxins. Results from the T4 model system provided a convincing demonstration that topoisomerase is the physiological drug target and strong evidence that the drug-induced cleavage complex is important for cytotoxicity. The detailed molecular steps involved in cytotoxicity, and the mechanism of recombinational repair of inhibitor-induced DNA damage, are currently being analyzed using this model system. Studies with the T4 topoisomerase have also provided compelling evidence that topoisomerase inhibitors interact with DNA at the active site of the enzyme, with each class of inhibitor favoring a different subset of cleavage sites based on DNA sequence. Finally, analysis of drug-resistance mutations in the T4 topoisomerase have implicated certain regions of the protein in drug interaction and provided a strong link between the mechanism of action of the antibacterial quinolones, which inhibit DNA gyrase, and the various antitumor agents, which inhibit mammalian type II topoisomerase. PMID- 9748649 TI - Protein kinase activities in ripening mango, Mangifera indica L., fruit tissue. III. Purification and characterisation of a calcium-regulated protein kinase. AB - A calcium-dependent protein kinase (PK-III), not requiring calmodulin for activity, was purified from extracts of ripening mango fruit tissue. Purification was achieved by ammonium sulfate fractionation and sequential anion exchange-, hydrophobic interaction-, dye ligand affinity- and gel filtration chromatography; which allowed a recovery of 1-5% of the total available kinase activity. The final specific activity in the presence of 1 mM Ca2+ was consistently 9 nmol min 1mg-1. The purified enzyme was a monomer with a Mr of 49000, but was resolved by denaturing electrophoresis into two related protein bands of 49 and 45 kDa. Enzyme activity was activated >30-fold by micromolar amounts of free calcium and was dependent upon millimolar Mg2+ or Mn2+ concentrations. Calmodulin (1 microM) had no effect on PK-III activity but the calmodulin antagonists, calmidazolium and chlorpromazine, inhibited PK-III in a dose-dependent manner over a range of 0 to 100 microM. The results suggest a regulatory domain that is similar to calmodulin. PK-III phosphorylated histone III-S and to a lesser extent casein, but did not phosphorylate histone II-S, phosvitin or protamine sulfate. The enzyme phosphorylated substrate proteins on either serine or threonine but not tyrosine. Some endogenous substrates and the ability to autophosphorylate were revealed by autoradiographic studies. PK-III displayed a broad pH optimum (pH 6.6 9.5), and the optimum reaction temperature with histone III-S as substrate was 35 degreesC. The kinetic reaction mechanism of PK-III was studied by using casein as substrate. The KmATP and Kmcasein of PK-III were determined as 10 microM and 1.0 mg ml-1, respectively. PMID- 9748650 TI - Dietary hydroperoxides of linoleic acid decompose to aldehydes in stomach before being absorbed into the body. AB - Our previous study (Biochim. Biophys. Acta 1393 (1998) 336-348, this issue) found that dietary hydroperoxides of trilinoleoylglycerol were broken down, releasing linoleic acid hydroperoxides (LA-OOH) in the stomach without reaching the intestines. The present paper describes the catabolic fate of LA-OOH in rat gastrointestines, in an attempt to elucidate those products which can be absorbed into the body. At an intragastric dose of 6.5 or 18 mumol, LA-OOH was not transported to the intestines as determined by HPLC. At large doses (200 or 800 mumol), much greater than that in the daily diet, there was partial leakage of LA OOH to the intestines. The periodical fate was analyzed with 17.2 mumol [U-14C]LA OOH chemically and radiochemically. Exemplifying the product composition at 30 min after treatment (as percentage of dosed amount), 27% unchanged LA-OOH, 9.7% epoxyketones, 3.5% hydroxyls (LA-OH), 2.4% decomposed aldehydes, and 13% unknown products were found in the gastric lumen. Another 25% was incorporated in the gastric tissue, and the other 6.4% occurred in the intestinal lumen and tissue as decomposed aldehyde. The LA-OH further decomposed to aldehydes with time in the stomach. When an aldehyde mixture was prepared and dosed, significant increases in hexanal and 4-hydroxynonenal were detected in the liver 15 h later. These results show that the dietary LA-OOH is decomposed to aldehydes in the stomach and that aldehydes are partly absorbed into the body. PMID- 9748651 TI - DNA topoisomerases: a new twist for antiparasitic chemotherapy? AB - The parasitic protozoa are notorious for their bizarre cellular structures and metabolic pathways, a characteristic also true for their nucleic acids. Despite these florid differences from mammalian cells, however, it has proven surprisingly difficult to devise novel chemotherapy against these pathogens. In recent years, the DNA topoisomerases from parasites have been the focus of considerable study, not only because they are intrinsically interesting, but also because they may provide a target for much-needed new antiparasitic chemotherapy. PMID- 9748653 TI - The amino acid sequences of carboxypeptidases I and II from Aspergillus niger and their stability in the presence of divalent cations. AB - The amino acid sequences of serine carboxypeptidase I (CPD-I) and II (CPD-II), respectively, from Aspergillus niger have been determined by conventional Edman degradation of the reduced and vinylpyridinated enzymes and peptides hereof generated by cleavage with cyanogen bromide, iodobenzoic acid, glutamic acid cleaving enzyme, AspN-endoproteinase and EndoLysC proteinase. CPD-I consists of a single peptide chain of 471 amino acid residues, three disulfide bridges and nine N-glycosylated asparaginyl residues, while CPD-II consists of a single peptide chain of 481 amino acid residues, has three disulfide bridges, one free cysteinyl residue and nine glycosylated asparaginyl residues. The enzymes are closely related to carboxypeptidase S3 from Penicillium janthinellum. Both Ca2+ and Mg2+ stabilize CPD-I as well as CPD-II, at basic pH values, Ca2+ being most effective, while the divalent ions have no effect on the activity of the two enzymes. PMID- 9748652 TI - Periplasmic and fimbrial SefA from Salmonella enteritidis. AB - Salmonella enteritidis produces thin, filamentous fimbriae composed of the fimbrin subunit SefA. Although insoluble in most detergents and chaotropic agents, these fimbriae were soluble at pH 10.5. Furthermore, in sodium dodecyl sulfate, these fibers depolymerized into monomers, dimers and other multimers of SefA, which precipitated on removal of the detergent. In contrast, unassembled periplasmic SefA fimbrins purified from Escherichia coli expressing cloned sefA and sefB were readily soluble in aqueous solution. Fimbrial and periplasmic SefA also differed in their reaction with an anti-SEF14 monoclonal antibody and in their surface hydrophobicity, indicating that the two forms had different properties. Precise mass measurements of periplasmic and fimbrial SefA by mass spectroscopy showed that these variations were not due to post-translational modifications. Periplasmic SefA consisted primarily of intact as well as some N terminally truncated forms. The main 24 amino acid, N-terminally truncated form of periplasmic SefA was present as a 12.2 kDa monomer which had a low tendency to dimerize whereas intact periplasmic SefA was present as a 34.1 kDa homodimer. Intact periplasmic SefA also formed stable multimers at low concentrations of chemical cross-linker but multimerization of the truncated form required high concentrations of protein or cross-linker. Thus, SefA fimbrins appear to multimerize through their N-termini and undergo a conformational change prior to assembly into fibers. Within these fibers, subunit-subunit contact is maintained through strong hydrophobic interactions. PMID- 9748654 TI - Specificity of Pseudomonas aeruginosa serralysin revisited, using biologically active peptides as substrates. AB - The characterization of the specificity of alkaline protease from Pseudomonas aeruginosa has not yet been clearly defined. Some previous results suggested that its specificity was influenced more by amino acids far from the hydrolyzed peptide bond, than by amino acids in P1 or P'1 position. From other data, it was a C-(COOH)-type endoprotease where the preferential amino acid in P1 position was an arginine residue. We have studied the hydrolysis of several biologically active peptides. Many various sites of cleavage have been characterized but no arginine in P1 position was found, despite the presence of arginine in the peptide sequence. In fact P1 and P'1 position could be occupied by various amino acids. It seems unlikely that Pseudomonas alkaline protease may only be considered as a protease specific to arginine in P1 position. On the other hand, we have observed that increase of the peptide chain length led to an important increase of the hydrolysis rate, suggesting an extended number of subsites. PMID- 9748655 TI - Phosphorylation of Mycobacterium leprae heat-shock 70 protein at threonine 175 alters its substrate binding characteristics. AB - We have examined the functional properties including autophosphorylation of the Mycobacterium leprae Hsp70 homologue. Recombinant M. leprae Hsp70 had pH optima for its adenosine triphosphatase and autophosphorylating activities which were near pH 8 and 6, respectively. Both these activities were inhibited by reduced and alkylated bovine pancreatic trypsin inhibitor, but not other tested substrates. Autophosphorylation was augmented by up to 25 mM Ca2+. Using site directed mutagenesis to construct two Thr-->Ala mutants at positions 175 and 193, and phosphoamino acid analysis, it was shown that Thr175 was the dominant threonine residue autophosphorylated in M. leprae Hsp70. Phosphorylation led to an increased affinity for a model polypeptide substrate, reduced and alkylated bovine albumin. These properties are compared with those of the DnaK protein of Escherichia coli. PMID- 9748656 TI - Steady-state kinetic mechanism, stereospecificity, substrate and inhibitor specificity of Enterobacter cloacae nitroreductase. AB - Enterobacter cloacae nitroreductase (NR) is a flavoprotein which catalyzes the pyridine nucleotide-dependent reduction of nitroaromatics. Initial velocity and inhibition studies have been performed which establish unambiguously a ping-pong kinetic mechanism. NADH oxidation proceeds stereospecifically with the transfer of the pro-R hydrogen to the enzyme and the amide moiety of the nicotinamide appears to be the principal mediator of the interaction between NR and NADH. 2,4 Dinitrotoluene is the most efficient oxidizing substrate examined, with a kcat/KM an order of magnitude higher than those of p-nitrobenzoate, FMN, FAD or riboflavin. Dicoumarol is a potent inhibitor competitive vs. NADH with a Ki of 62 nM. Several compounds containing a carboxyl group are also competitive inhibitors vs. NADH. Yonetani-Theorell analysis of dicoumarol and acetate inhibition indicates that their binding is mutually exclusive, which suggests that the two inhibitors bind to the same site on the enzyme. NAD+ does not exhibit product inhibition and in the absence of an electron acceptor, no isotope exchange between NADH and 32P-NAD+ could be detected. NR catalyzes the 4-electron reduction of nitrobenzene to hydroxylaminobenzene with no optically detectable net formation of the putative two-electron intermediate nitrosobenzene. PMID- 9748657 TI - Conformational changes of the leukocyte NADPH oxidase subunit p47(phox) during activation studied through its intrinsic fluorescence. AB - The leukocyte NADPH oxidase of neutrophils is a membrane-bound enzyme that catalyzes the production of O-2 from oxygen using NADPH as the electron donor. Dormant in resting neutrophils, the enzyme acquires catalytic activity when the cells are exposed to appropriate stimuli. During activation, the cytosolic oxidase components p47phox and p67phox migrate to the plasma membrane, where they associate with cytochrome b558, a membrane-integrated flavohemoprotein, to assemble the active oxidase. Oxidase activation can be mimicked in a cell-free system using an anionic amphiphile, such as sodium dodecyl sulfate or arachidonic acid, as an activating agent. In whole cells and under certain circumstances in the cell-free system the phosphorylation of p47phox mediates the activation process. It has been proposed that conformational changes in the protein structure of cytosolic factor p47phox may be an important part of the activation mechanism. We show here that the total protein steady-state intrinsic fluorescence (an emission maximum of 338 nm) exhibited by the tryptophan residues of p47phox substantially decreased when p47phox was treated with anionic amphiphiles. A similar decrease in fluorescence was also observed when p47phox was phosphorylated with protein kinase C. Furthermore, a red shift of emission maximum and an increase of quenching by ionic quenchers and acrylamide were observed in the presence of activators. These results indicate the occurrence of a conformational change in the protein structure of p47phox. We propose that this alteration in conformation results in the appearance of a binding site through which p47phox interacts with cytochrome b558 during the activation process. PMID- 9748658 TI - Interaction of second and third domains of Japanese quail ovomucoid with ten mammalian trypsins. AB - Second and third domains were prepared from Japanese quail ovomucoid and association equilibrium constants, Kas, were measured at 25 degreesC and pH 8 for these domains with trypsins from ten mammalian species: cat, cow, dog, guinea pig, hog, horse, man, rabbit, rat, and sheep. The values ranged from 108 M-1 to 1010 M-1 for the second domain-trypsin associations and from 106 M-1 to 108 M-1 for the third domain-trypsin associations. Changes in Ka values for the interactions between the trypsins and each domain are attributed to slight changes in surface conformation caused by the residue changes in the inhibitor binding region other than the S1 pocket of the trypsin species. The representative of such residue changes is assumed to be the one observed at residue 217 of trypsin molecule. Concerning each trypsin, the Ka value with the second domain was always higher than that with the third domain. However, the ratios between the two equilibrium constants varied from 3 to 60 depending upon trypsin species. This means that amino acid changes in enzyme-contact residues other than the P1 site of the Kazal-type inhibitor can make it possible to recognize even a slight difference in inhibitor-binding surface among the enzymes with the same S1 pocket and highly similar overall three-dimensional structure. PMID- 9748659 TI - Protein trans-splicing and functional mini-inteins of a cyanobacterial dnaB intein. AB - A 429 aa theoretical intein is encoded in the dnaB gene (DNA helicase) of the cyanobacterium Synechocystis sp. strain PCC6803. This intein is shown to be capable of protein splicing with or without its native exteins when tested in E. coli cells. A centrally located 275 amino acid sequence (residues 107-381) of this intein can be deleted without loss of the protein splicing activity, resulting in a functional mini-intein of 154 aa in size. Efficient in vivo protein trans-splicing was observed when this mini-intein was split into a 106 aa N-terminal fragment containing intein motifs A and B, and a 48 aa C-terminal fragment containing intein motifs F and G. These results indicate that the N- and C-terminal regions of the Ssp DnaB intein, whether covalently linked with each other or not, can come together through non-covalent interaction to form a protein splicing domain that is functionally sufficient and structurally independent from the centrally located endonuclease domain of the intein. PMID- 9748660 TI - Site-directed mutagenesis of surfactant protein A reveals dissociation of lipid aggregation and lipid uptake by alveolar type II cells. AB - Surfactant protein A (SP-A) binds to dipalmitoylphosphatidylcholine (DPPC) and induces phospholipid vesicle aggregation. It also regulates the uptake and secretion of surfactant lipids by alveolar type II cells. We introduced the single mutations Glu195-->Gln (rE195Q), Lys201-->Ala (rK201A) and Lys203-->Ala (rK203A) for rat SP-A, Arg199-->Ala (hR199A) and Lys201-->Ala (hK201A) for human SP-A, and the triple mutations Arg197, Lys201 and Lys203-->Ala (rR197A/K201A/K203A) for rat SP-A, into cDNAs for SP-A, and expressed the recombinant proteins using baculovirus vectors. All recombinant proteins avidly bound to DPPC liposomes. rE195Q, rK201A, rK203A, hR199A and hK201A function with activity comparable to wild type SP-A. Although rR197A/K201A/K203A was a potent inducer of phospholipid vesicle aggregation, it failed to stimulate lipid uptake. rR197A/K201A/K203A was a weak inhibitor for lipid secretion and did not competed with rat [125I]SP-A for receptor occupancy. From these results, we conclude that Lys201 and Lys203 of rat SP-A, and Arg199 and Lys201 of human SP-A are not individually critical for the interaction with lipids and type II cells, and that Glu195 of rat SP-A can be replaced with Gln without loss of SP-A functions. This study also demonstrates that the SP-A-mediated lipid uptake is not directly correlated with phospholipid vesicle aggregation, and that specific interactions of SP-A with type II cells are involved in the lipid uptake process. PMID- 9748661 TI - Subunit conformation and dynamics in a heterodimeric protein: studies of the hybrid isozyme of creatine kinase. AB - Several physical properties of creatine kinase (EC 2.7.3.2) isozymes MM (CK-MM, muscle-type) and BB (CK-BB, brain-type), both homodimers, and isozyme MB (CK-MB), a heterodimer, were compared to determine how formation of the hybrid modifies subunit conformation and dynamics. Circular dichroic spectra revealed additional alpha-helical content for the hybrid isozyme. Double-beam absorption difference spectra between CK-MB and a stoichiometric mixture of CK-MM and CK-BB revealed decreased exposure of intrinsic chromophores in the hybrid. The relative intensity of the intrinsic fluorescence of CK-MB was between the two homodimers, but was 16% closer to the less fluorescent CK-MM. Steady state anisotropy spectra and decay of the anisotropy of CK derivatized on a single subunit with the fluorescent sulfhydryl reagent 5-[2-(iodoacetyl)amino-ethyl]aminonaphthalene-1 sulfonate indicated that the derivatized sites are more flexible in the heterodimer. The slow component in the anisotropy decay suggests that hybridization results in a small increase in the packing density or contraction of overall conformation of the B-subunit. The KM for MgATP with singly derivatized CK-MB was the same as the KM for the native enzyme. However, derivatization of a single subunit caused the Vmax to decrease by greater than 50%, which indicates that subunit-subunit interactions may modulate the activity of CK. A model for assembly of CK-MB is proposed which includes subunit characteristics more similar to those found in the muscle-type homodimer than in the brain-type homodimer and increased flexibility of the active site domain of both subunits. PMID- 9748662 TI - The five cysteine residues located in the active site region of bovine aspartyl (asparaginyl) beta-hydroxylase are not essential for catalysis. AB - In previous chemical modification studies on bovine aspartyl (asparaginyl) beta hydroxylase, cysteines were implicated as critical catalytic residues. Using site directed mutagenesis, the five cysteine residues located in a highly conserved region of the enzyme identified as the active site were individually mutated to alanine. Substitutions at cysteine 637, 644, 656, 681, and 696 resulted in active mutant enzymes indicating that these residues are not required for catalysis. PMID- 9748663 TI - Molecular cloning and sequence determination of a novel aspartic proteinase from Antarctic fish. AB - In the present report, we describe a novel aspartic proteinase from the liver of two Antarctic fish species. The nucleotide sequences of the cDNA obtained from the two fishes show 90% identity with each other but only 58% identity with aspartic proteinases from other sources. Sequence analysis shows features for the Antarctic enzymes which are not present in related enzymes of other organisms. PMID- 9748664 TI - Carbamoyl-phosphate synthetase II in kinetoplastids. AB - Genes for carbamoyl-phosphate synthetase II (CPS II), the first enzyme of de novo pyrimidine biosynthesis, were cloned from kinetoplastids, Trypanosoma cruzi and Leishmania mexicana. T. cruzi CPS II gene encodes a protein of 1524 amino acids that encompasses the glutaminase and CPS domains, but incorporates neither aspartate carbamoyltransferase nor dihydroorotase. The residue corresponding to lysine 993 of Escherichia coli CPS, a residue that characterizes the CPS inhibited by UMP and that is replaced by tryptophan in those inhibited by UTP, is in kinetoplastids a hydrophilic glutamine, in line with the preferential inhibition by UDP of kinetoplastid CPS II. PMID- 9748665 TI - Genomic DNA sequence of a rice gene coding for a pullulanase-type of starch debranching enzyme. AB - A genomic DNA containing a rice (Oryza sativa L., cv. Norin-8) gene coding for a pullulanase-type starch debranching enzyme (EC 3.2.1. 41) was sequenced (EMBL/GenBank/DDBJ accession number AB012915). Along the 15, 248 bp DNA, the pullulanase gene is split into 26 exons. The four pullulanase consensus regions are positioned in the middle portion of the sequence and are separated by long introns and 1-3 exons. Comparison of the rice cv. Norin-8 pullulanase genomic structure with that of barley pullulanase (limit dextrinase) (F. Lok et al., EMBL/GenBank/DDBJ accession number AF022725) indicates that most of the pullulanase exons are highly conserved. Alignment of the nucleotide bases of rice exon 8 with those of barley exon 8-intron 8-exon 9 fragment suggests that the 85 bp internal sequence of rice exon 8 was originally an intron, a possibility further indicated by the absence in barley and spinach (A. Renz et al., EMBL/GenBank/DDBJ accession number X83969) pullulanases of amino acid residues encoded by the 85 bp fragment. PMID- 9748666 TI - Heterogeneity of Atlantic salmon troponin-I. AB - Three non-identical, full length troponin-I (Tn-I) clones were isolated from an Atlantic salmon myotomal (trunk) muscle cDNA library. The primary structures, which are predicted to range from 172 to 180 amino acids in length, exhibit similar percent identity scores when compared with fast, slow and cardiac specific Tn-Is from higher vertebrates. When the sequence data are considered along with the results of Western blotting it is evident that Tn-I is more heterogeneous in Atlantic salmon than has been previously shown in higher vertebrates. PMID- 9748667 TI - Corrigendum to: hatching enzyme from the sea-squirt ciona intestinalis: purification and properties. PMID- 9748669 TI - Re-evaluating the role of vascular changes in the differential diagnosis of Alzheimer's disease and vascular dementia. AB - Alzheimer's disease (AD) and vascular dementia (VaD) are the two most common causes of dementia, and much effort has been devoted to their differential diagnosis. However, current epidemiological, clinical and neuropathological evidence points to a substantial overlap between AD and VaD and suggests that vascular pathology, the traditional cornerstone of the differential diagnosis between the two entities, may not represent as clear a line of demarcation as originally believed. It may be time to reevaluate the dichotomy between AD and VaD. PMID- 9748670 TI - Neurofibrillary tangles and Alzheimer's disease. AB - The neuropathological diagnosis of Alzheimer's disease relies on the presence of both neurofibrillary tangles and senile plaques. The number of neurofibrillary tangles is tightly linked to the degree of dementia, suggesting that the formation of neurofibrillary tangles more directly correlates with neuronal dysfunction. The regional pattern of areas affected by neurofibrillary tangles formation during the course of the disease is relatively stereotyped. Neurofibrillary tangles are composed of highly phosphorylated forms of the microtubule-associated protein tau. Phosphorylated tau proteins accumulate early in neurons, even before formation of neurofibrillary tangles, suggesting that an imbalance between the activities of protein kinases and phosphatases acting on tau is an early phenomenon. The latter might be related to changes in signalling through transduction cascades, since many of the protein kinases generating phosphorylated tau species participate in signalling pathways. The accumulation of neurofibrillary tangles and phosphorylated tau species is associated with disturbances of the microtubule network and, as a consequence of the latter, of axoplasmic flows. The mechanistic relationship between the formation of neurofibrillary tangles and senile plaques is still little understood and in vivo formation of neurofibrillary tangles in experimental models has not yet been achieved. Future animal models, e.g. transgenic animals expressing combined key human proteins, will hopefully reproduce faithfully all the major cellular lesions of the disease. PMID- 9748671 TI - A unique autosomal dominant disorder with indifference to pain: clinicopathologic correlation of indifference to pain and thalamic gliosis. AB - We present updated information on a previously reported kindred with an autosomal dominant disorder variably expressed as indifference to pain, dementia, and ataxia. Additional clinical and radiological information is presented, as are autopsy results form the index case. In addition to evidence of Alzheimer's disease, the autopsy revealed bilateral thalamic gliosis, which may be a neuroanatomic substrate for the indifference to pain seen in this patient. To our knowledge, this is the first reported association of thalamic gliosis and indifference to pain. PMID- 9748672 TI - Detecting new lesion formation in multiple sclerosis: the relative contributions of monthly dual-echo and T1-weighted scans after triple-dose gadolinium. AB - In this study, we evaluated the frequency of formation of new lesions on brain magnetic resonance imaging (MRI) from patients with relapsing-remitting multiple sclerosis (MS) and defined the relative contributions of unenhanced and enhanced MRI. Every 4 weeks for 3 months, dual-echo and postcontrast T1-weighted (5 min after the injection of 0.3 mmol/kg gadolinium-DTPA) scans were obtained from 28 patients with relapsing-remitting MS. New lesions were defined as those present on dual-echo and/or postcontrast T1-weighted scans but with no corresponding MRI abnormalities on any of the preceding scans. A total of 111 newly formed lesions were detected during the follow-up on dual-echo and postcontrast T1-weighted scans (i.e., an average of 1.3 lesions per patient per month). Ninety-eight (88%) of such lesions were seen on both dual-echo and postcontrast T1-weighted scans, whilst 13 (12%) were seen only with one of the two techniques: 9 only on dual echo and 4 only on postcontrast scans. Five of the 98 new lesions seen by both techniques were seen by postcontrast scans 1 month before their appearance on dual-echo scans. Our study suggests that both dual-echo and postcontrast T1 weighted scans are useful to detect newly formed lesions in patients with MS. This is of importance when using MRI to monitor the efficacy of treatments which may halt MS lesion formation. PMID- 9748673 TI - Contribution of the supplementary motor area and anterior cingulate gyrus to pathological grasping phenomena. AB - To investigate the relationship between the site of brain damage and characteristics of the pathological grasping phenomena, we examined different varieties of the reaction in a consecutive series of 28 patients with unilateral hemispheric damage due to stroke. Patients with a lesion relatively confined to the supplementary motor area (n = 4) constantly exhibited a grasp reflex, mainly in the hand contralateral to the lesion, but they never showed a groping reaction. By contrast, patients with damage primarily involving the anterior cingulate gyrus (n = 3) developed the groping reaction in the hand contralateral to the lesion, but they had only a very mild grasp reflex in that hand. Patients with damage involving both the supplementary motor area and the anterior cingulate gyrus (n = 12) showed the grasp reflex and groping reaction mainly in the hand contralateral to the lesion. Patients with damage to the medial parietal lobe (n = 2), those with damage to the lateral convexity of the hemisphere (n = 6), and a patient with damage confined to the corpus callosum did not exhibit such grasping phenomena. From these observations, we conclude that the grasp reflex is closely related to a lesion of the supplementary motor area, whereas the groping reaction is bound to a lesion of the anterior cingulate gyrus. PMID- 9748674 TI - Indicators and prevalence of malnutrition in motor neurone disease. AB - We investigated the nutritional status of a cross-sectional sample of 47 patients with motor neurone disease (MND). Anthropometric measurements (mid-arm circumference and triceps skinfold thickness) and dietary analysis were taken by a trained observer. 21% of patients were moderately to severely malnourished. Patients with bulbar onset were no more likely to be malnourished than patients with limb onset. Measurements of mid-upper-arm muscle circumference indicated that men were more malnourished than women (p = 0.002). We conclude that malnutrition is more prevalent than generally appreciated in patients with MND, including those who do not have swallowing difficulties. PMID- 9748675 TI - CAG repeat expansion in an italian family with spinocerebellar ataxia type 2 (SCA2): a clinical and genetic study. AB - We report an Italian family in which molecular genetic analysis showed expansion of CAG repeats indicative of the SCA2 genotype. This family confirms that ataxia, ophthalmoparesis and sensory peripheral neuropathy are the salient features of the SCA2 phenotype. In the present cases, early onset and mental deterioration were important additional findings. Nerve biopsy findings were compatible with a chronic axonopathy. We found a direct correlation between length of triplet expansion and severity of the clinical symptoms. Of particular interest is the late-onset phenotypical expression in a patient with 34 repeats. PMID- 9748677 TI - Considerations regarding prostate biopsies. AB - OBJECTIVES: To review the current clinical practice concerning prostate biopsies and indications for prostate biopsies and to study the value of biopsies in grading and staging of prostate cancer. METHODS: The literature from 1990 onwards was reviewed systematically. A selection out of the huge number of publications concerning the subject was made based on the relevance of the study (e.g., number of patients, study design). RESULTS: Transrectal ultrasound-guided biopsies have become a routine procedure in urological practice and can be performed safely. Antibiotic prophylaxis is recommended generally. Sextant biopsies should be performed when no lesion is visible or added to lesion-directed biopsies in case of a visible lesion. The indications for biopsies and for repeat biopsies are discussed. The indication for biopsies remains a problem in spite of prostate specific antigen and prostate specific antigen derived indexes and in spite of new imaging techniques. The value of prostate biopsy in grading and staging is limited, and care should be taken not to base treatment decisions on prostate biopsy results only and not to compare treatment results based on biopsy data. CONCLUSIONS: Prostate biopsies have become a routine procedure in urology. Although very helpful in many cases, their limitations should be kept in mind. More efforts will have to be made to reduce the (too) large number of negative biopsies by improvement of imaging techniques and development of more sensitive and specific tumor markers. PMID- 9748676 TI - Acute disseminated encephalomyelitis following Pontiac fever. AB - We report the case of a 35-year-old woman who developed headache and psychosis and gradually became comatose within 3 weeks after a flu-like infection. MRI revealed bifrontal demyelination consistent with acute disseminating encephalomyelitis (ADEM). Two different cerebrospinal fluid samples were positively tested for Legionella cincinnatiensis by direct sequencing of a PCR amplified Legionella-specific fragment. This result made it possible to interpret the initial symptoms as Pontiac fever. We think it most likely that this is a case of ADEM following the very rare situation of a systemic infection with L. cincinnatiensis. A review of the literature on Legionella-associated encephalopathy suggests that some of these cases may also have had ADEM. PMID- 9748678 TI - Use of three additional mid biopsies to improve local assessment of prostate cancer in patients with one positive sextant biopsy. AB - OBJECTIVES: Routine sextant biopsies have proven useful in the diagnosis and local staging of prostate cancer. A single positive biopsy is more frequently associated with a smaller tumor and a low risk of positive margins. Nevertheless, the risk of positive margins in patients with 1 positive sextant biopsy remains high (20%). A better assessment of local invasion is therefore needed. In addition to standard sextant biopsies, we routinely obtain 3 additional mid biopsies from the apex to the base of the prostate. The aim of this study is to analyze the contribution of these 3 additional mid biopsies to local staging. METHODS: From 1988 to 1996, 177 men underwent sextant biopsies plus 3 additional mid biopsies prior to radical prostatectomy; 59 men had 1 positive sextant biopsy, and 13 also had 1-3 positive mid biopsies. The pathological results of the prostatectomy specimens from these 13 men (group A) were compared with those of the 46 men with only 1 positive sextant biopsy (group B), by means of the Fisher and Mann-Whitney tests. RESULTS: The two groups were similar in terms of age, preoperative prostate-specific antigen, the Gleason score of positive biopsies, the weight of the specimen, the Gleason specimen score, tumor volume and pathological stage. Positive surgical margins were found in 53.8% of group A and 19.4% of group B patients (p = 0.002). The location of the positive additional biopsies matched that of the positive surgical margins. CONCLUSIONS: Additional mid biopsies improve the local assessment of prostate cancer in patients with a single positive sextant biopsy, identifying significantly more positive margins when these additional mid biopsies are positive and indicating the location of the positive surgical margins. These informations could be helpful to avoid positive surgical margins during radical prostatectomy. PMID- 9748679 TI - Supersensitive PSA-monitored neoadjuvant hormone treatment of clinically localized prostate cancer: effects on positive margins, tumor detection and epithelial cells in bone marrow. AB - OBJECTIVE: The present study was done to investigate the effects of supersensitive PSA-controlled inductive treatment on positive margins, detection of tumor and epithelial cells in bone marrow of 101 patients with untreated and clinically localized prostatic carcinoma (cT1-3N0M0). METHODS: Hormonal treatment was given until PSA (DPD Immulite(R) third-generation assay) reached <0.1 ng/ml or the nadir value, as shown by two consecutive measurements at monthly intervals. RESULTS: The resultant median duration of treatment was 6 months (range 3-22). Ninety-three (93%) of our patients reached a PSA value <0.1 ng/ml. The nadir of 6 patients (6%) was between 0.1 and 0.3 ng/ml, and it remained >0.3 ng/ml in only 1 case. Of the 101 patients, 82 had a measurable hypoic lesion on initial transrectal ultrasound. 84% of these became smaller, 7.5% remained unchanged and 8.5% increased. Of the 101 prostatectomy specimens, 20 (20%) were margin-positive. The incidence of affected margins was relatively high (35% from 55 patients) with cT3 tumors, but almost negligible (2% from 46 patients) in cT1 2 tumor. Our pathologists, despite their great experience in evaluating hormonally treated prostates (>500 cases) and using immunohistochemical staining, were unable to detect carcinoma in 15 (15%) specimens. Whereas only 2 (4%) of the 55 cT3 specimens were without detectable tumor, this incidence rised to 28% (13 of 46 prostates) in patients with cT1-2 tumors. Of the initial 29 patients with epithelial cells in bone marrow, only 4 (14%) remained positive after controlled induction and all of them had fewer cells than before. CONCLUSION: Endocrine induction controlled by a supersensitive PSA assay and continued until reaching PSA nadir is highly effective in clearing surgical margins and eliminating tumor cells from bone marrow. It seems to be clearly superior to the conventional 3 months of pretreatment at least in cT1-2 tumors in respect to surgical margins and detectability of tumor in the resected prostate. A definitive statement about the value of endocrine induction can only be given by prospective randomized studies, with optimal drugs, doses and treatment time. But the conventional 3 months of pretreatment are far from exploiting the possibilities of this therapeutic option. PMID- 9748680 TI - The natural history of untreated lower urinary tract symptoms in middle-aged and elderly men over a period of five years. AB - OBJECTIVES: To describe changes in untreated lower urinary tract symptoms (LUTS), levels of bothersomeness and interference in selected living activities arising from LUTS in a cohort of men followed up for a period of 5 years. METHODS: 1,994 men aged 40-79 years registered in four health centres in Scotland completed a urinary symptom questionnaire in the community. 1,177 (71% of original eligible participants) were followed up at 5 years. RESULTS: The mean level of LUTS and the level of bothersomeness they caused increased universally between baseline and 5 years. The largest increases in mean symptom levels occurred for straining, hesitancy, incomplete emptying, weak stream and nocturia, and for mean levels of bothersomeness, i.e. dribbling, incomplete emptying, weak stream, frequency, nocturia and intermittency. The increase in mean interference level in selected living activities arising from LUTS was less marked. The progression of LUTS over time in individual men was very variable; however, the overall trend was one of continuing deterioration. CONCLUSION: If left untreated, overall levels of middle aged and elderly mens' LUTS, bothersomeness and interference in selected activities caused by these symptoms will increase over time. PMID- 9748682 TI - The Stamey procedure for stress incontinence: long-term results. AB - OBJECTIVE: To assess the long-term results of the Stamey vesicourethral suspension for stress urinary incontinence. METHODS: A total of 72 case records of women who underwent the Stamey procedure in the years 1985-1991 was reviewed. Every patient had a full preoperative evaluation including filling cystometry, urethral pressure profilometry and imaging of the kidneys and bladder. The long term subjective results were determined by a telephone interview with the patients, utilizing a standard questionnaire. A successful result was defined as no need for pads under any circumstances. Every patient who reported on persistent or recurrent incontinence was requested to return for reevaluation in the urology clinic. RESULTS: The long-term success rate of the Stamey procedure in 63 evaluable patients was 69.8% (mean follow-up: 90 months). In 19 women, the operation had failed. About 80% of the failures were evident within 2 years after surgery. The long-term success rate of surgery in 28 women with pure stress incontinence was 93%. When mild irritative symptoms were present before surgery (29 patients) the subjective success rate was 65.5%, and in 6 patients with severe urge symptoms the success rate was only 33%. The difference between these groups was highly significant (p = 0.003). No correlation was found between the age of the patient at surgery, the number of children delivered by the patient, the grade of stress incontinence, the duration of symptoms before surgery or a history of a previous operation for stress incontinence and the success rate of the operation. CONCLUSIONS: The Stamey vesicourethral suspension is characterized by a relatively high and long-standing success rate. This procedure has an important place in the treatment of women with urinary stress incontinence. PMID- 9748681 TI - Prevalence of urinary incontinence among Spanish older people living at home. AB - OBJECTIVE: To estimate the prevalence and characteristics of urinary incontinence (UI) in the noninstitutionalized elderly population of Madrid, Spain. METHODS: We carried out a cross-sectional study in a representative sample of all community dwelling people aged 65 or over. Subjects were interviewed in their homes. The question: Do you currently experience any difficulty in controlling your urine? . In other words, does your urine escape involuntarily? was used to identify UI. Type of UI, use of absorbents and specific drugs were also assessed, as well as consultation behavior. RESULTS: 589 persons were interviewed (response rate: 71.2%). The prevalence of UI was 15.5%. No significant difference was observed between men and women. Urge UI was the main type for men and mixed UI for women. Use of pads was referred by 20.2%. A total of 34.3% of subjects never went to the doctor for their problem (25.2% of men and 39.4% of women). CONCLUSION: Compared to other populations the overall prevalence of UI in Spanish elders living at home is relatively high. A very small difference by gender was found, although a lower response rate in women could in part explain this unexpected finding. PMID- 9748683 TI - Ureteroscopy in the treatment of ureteral stones: over 10 years' experience. AB - OBJECTIVE: Rapid development of endourology and the invention of more and more advanced ureteroscopes and other instruments used in ureteral lithotripsy have made the traditional methods of treatment become very rare. METHODS: We present our experience in ureteral lithotripsy resulting from 1,982 ureteroscopy (URS) procedures, performed because of ureteral stones. Before URS, percutaneous nephrostomy tube (PCNT) was created in 264 (16.7%) cases. We also present our own technique, called the 'Jeromin maneuver', which involves pressing the abdominal wall by the assistant's hand, facilitating URS in difficult cases. RESULTS: Good results after the first URS procedure of removing ureteral stones were obtained in 1, 364 (86.6%) patients out of 1,575. In the remaining 211 (13.4%) cases, URS was performed two or more times. The overall failure rate was 3.6%. In the vast majority of cases, URS procedures were performed without dilatation of the ureteral orifice and splinting. The most important complications of URS were: perforation of the ureteral wall with periureteral leak which necessitated surgery (4 patients), ureteral stenosis which necessitated endoscopic reparation (4 patients) and stenosis of the ureteral orifice which necessitated endoscopic reparation in 2 patients. CONCLUSIONS: Routine dilatation of the ureteral orifice before the URS procedure and splinting with a D-J catheter are unnecessary; in case of a narrow ureter and very large prostatic adenoma, URS should not be attempted, because of the high risk of serious damage of the ureter. URS is a safe procedure but requires a highly experienced urologist. PMID- 9748684 TI - Prospective comparative study with intracavernous sodium nitroprusside and prostaglandin E1 in patients with erectile dysfunction. AB - PURPOSE: To compare the effectiveness of intracavernous administration of sodium nitroprusside and prostaglandin E1 to induce penile erection in men with erectile dysfunction. MATERIAL AND METHODS: 100 patients with erectile dysfunction entered the study prospectively. As part of the diagnostic workup, each patient received an intracavernous injection of 20 microg prostaglandin E1 and a second injection of 600 microg sodium nitroprusside 1-7 days later. A tourniquet was placed at the base of the penis before each injection. The data recorded included time required to initiate tumescence, local and systemic side effects, objective and subjective quality of erections, duration of tumescence and patient satisfaction by means of a personal questionnaire. RESULTS: Prostaglandin E1 induced better overall responses than sodium nitroprusside, the difference being almost significant (p = 0.055). The overall duration of erections was also significantly longer with prostaglandin E1 (mean 81.3 min) than with sodium nitroprusside (mean 65.4 min; p < 0.04). 67% of the patients considered the erections induced with prostaglandin E1 to be of better quality than those with sodium nitroprusside, and only 11% stated that sodium nitroprusside was superior. Side effects were minimal with both drugs, the most frequent side effect being systemic hypotension, which was induced by sodium nitroprusside in 7% of the patients. CONCLUSIONS: The moderate risk of systemic hypotension and the lower potency of sodium nitroprusside to induce erections compared to prostaglandin E1 rules out sodium nitroprusside as a routine alternative intracavernous drug in men with erectile dysfunction at the doses employed. Sodium nitroprusside, however, could be used in patients who have intolerance or penile pain with intracavernous prostaglandin E1. PMID- 9748685 TI - Treatment of erectile dysfunction by an external ischiocavernous muscle stimulator. AB - OBJECTIVE: The aim of the present study was to evaluate the therapeutic potency of an electrotherapy of striated ischiocavernous muscles in patients with erectile dysfunction. PATIENTS AND METHODS: Transcutaneous electrostimulation of striated ischiocavernous muscles by self-adhesive penile or perineal skin electrodes was performed in 48 patients with erectile dysfunction. 6/48 patients (R) responded to intracavernous pharmacotherapy while 42/48 (NR) did not show significant penile rigidity even to intracavernous papaverine/phentolamine/PGE1 triple drug medication. RESULTS: Within the observation time of 3 months, 10/48 patients dropped out. 22/38 patients reported a penile rigidity for sufficient sexual intercourse whereby 3/22 NR required additional intracavernous pharmacotherapy. Penile rigidity could be objectivated by triple drug medicaton in 12/14 NR after ischiocavernous muscle stimulation (EIS) therapy. 5/6 R were treated successfully for premature erection loss. During EIS treatment neither discomfort nor complications could be observed. CONCLUSION: Transcutaneous electrostimulation of ischiocavernous muscles is a new, noninvasive therapy for the improvement of penile rigidity. The clinical results underline the importance of the striated ischiocavernous muscles for penile rigidity. PMID- 9748686 TI - Detrusor overactivity and penile erection in patients with lower lumbar spine lesions. AB - OBJECTIVES: To investigate urodynamically detrusor overactivity and evaluate surgical outcome in patients with lower lumbar spine lesions. METHODS: Eighty patients with spine lesions below the third lumbar spine including 31 patients with intervertebral disc prolapse (IDP) (mean age: 38.2 years) and 49 patients with spinal canal stenosis (SCS) (mean age: 56.7 years) were studied. Urinary symptoms and urodynamic data were recorded before and after orthopedic surgery. RESULTS: Detrusor overactivity was noted in 17 patients: 3 IDP patients (10%) and 14 SCS patients (29%). Of these 17 patients (9 males and 8 females), 14 (82%) had voiding symptoms and 15 (88%) had storage symptoms. One patient was asymptomatic. Intermittent claudication was noted in 14 patients: 8 had urgency, 11 urge incontinence and 4 male patients showed erection on walking. Of 10 patients followed up after surgery, detrusor overactivity disappeared in 5 patients, improved in 1 patient and remained unchanged in 4 patients. CONCLUSIONS: Detrusor overactivity was found in lower lumbar spine lesions. This phenomenon seemed to be caused by the irritation of sacral roots, especially on walking. PMID- 9748687 TI - One-stage repair of hypospadias--experience with 856 cases. AB - OBJECTIVES: 856 cases of hypospadias, operated over a period of 7 years, using various methods of one-stage repair, are reviewed in order to assess their functional and cosmetic results and their rate of complications. PATIENTS AND METHODS: 856 boys with an average age of 3.5 years were classified according to the classification proposed by Duckett (anterior, middle and posterior hypospadias). Magpi, Mathieu's (flip-flap technique) and pedicled flaps tubularized or as onlay were utilized, depending upon the severity of the anomaly. RESULTS: Satisfactory functional and cosmetic results without complications were obtained in 736 cases (85.9%). We had early complications with 72 cases consisting of 64 fistulas (7. 5%) and 8 cases of breakdown (0.93%), and 48 cases of delayed complications consisting of 26 meatal retractions (3.1%) and 22 cases of meatal stenosis (2.5%). PMID- 9748688 TI - Surgical repair of anterior hypospadias with fish-mouth meatus and intact prepuce based on anatomical characteristics. AB - PURPOSE: A variant form of anterior hypospadias, called a megameatus and intact prepuce (MIP), is thought to be less amenable to conventional distal hypospadias repair. The feasibility of using the standard technique with a parameatal-based foreskin flap is described herein. MATERIALS AND METHODS: Nine children with the MIP variant underwent repair. A foreskin flap for urethroplasty was harvested from either the ventral (Mathiew) or unilateral site. The glans was split along with the cleft glanular groove to create the glans wings. The flap was laid on the urethral plate to form a neourethra, and glanulomeatoplasty was completed by approximation of the glans wings. Sleeve reapproximation of the penile foreskin was performed for uncircumcised skin closure. RESULTS: The functional and cosmetic results of the procedure were excellent in 8 cases including 1 with temporary postoperative edema of redundant foreskin. The last case underwent excision of the ventral excess foreskin for cosmetic reasons. CONCLUSIONS: Although the etiology of the MIP variant remains obscure, the urethral plate distal to the meatus is uniformly pliable and healthy in this variant. Furthermore, the ventral portion just proximal to the meatus is well developed and not atretic so that the parameatal ventral foreskin is safely harvested for onlay urethroplasty. PMID- 9748690 TI - DNA flow cytometry reveals depressed spermatogenetic activity in the contralateral testis within 24 h of ipsilateral spermatic cord torsion independently of the presence of the testis and epididymis. AB - An experimental study was planned to evaluate if contralateral testicular deterioration following ipsilateral torsion requires the presence of a twisted ipsilateral testis and/or epididymis. Five groups, each containing 6 rats, were established. The groups underwent sham operation, epididymo-orchiectomy, testicular torsion, torsion following subepididymal orchiectomy and torsion following epididymo-orchiectomy. After 24 h, the contralateral testes were harvested and the percentage of haploid cells was determined by DNA flow cytometry. Ipsilateral torsion, in the presence and absence of testis and epididymis, significantly decreased the number of haploid cells compared to sham and epididymo-orchiectomy procedures. Torsion of only the ipsilateral spermatic cord and adjacent vasculature seemed to suffice to damage the contralateral testis. Since the presence of the ipsilateral testis and epididymis is not mandatory for this acquired damage, which occurs within 24 h, a role for a preexisting congenital defect and autoimmunity seems unlikely. PMID- 9748689 TI - Histological evaluation of the effects of electropermeabilization after administration of bleomycin on bladder cancer in the rat. AB - OBJECTIVE: The aim of this study was to examine the usefulness of an electropermeabilization method in the administration of a chemotherapeutic agent in bladder cancer in the rat. MATERIALS AND METHODS: N-Butyl-N-(4-hydroxybutyl) nitrosamine was used to induce bladder tumors in Wistar rats. Rats in group 1 were given a sham treatment. Rats in group 2 received an injection of bleomycin (BLM). Rats in group 3 received a series of eight 100- micros square-wave pulses at an electric field intensity of 1,000 V/cm (electropermeabilization) applied to the bladder. Rats in group 4 were treated with square-wave pulses after the injection of BLM. Nine days after the various treatments, all the bladders were removed and examined histopathologically. RESULTS: Histopathological examination revealed severe damage to tumors in group 4 exclusively. CONCLUSION: The cytotoxic effect of BLM on bladder cancer tissue was enhanced by electropermeabilization. PMID- 9748692 TI - Repetitive exon 45/46-related sequences of human Ca2+ channel alpha1C subunit gene. AB - We found in the Ca2+ channel alpha1C subunit gene a new repetitive element of three paired exon 45/46-related sequences. We also identified a new exon 45/46 related sequence in the human genome and mapped it by fluorescence in situ hybridization to the 12p11.2 and 12p13.2-p13.1 bands. These positions are not recognized by DNA probes generated from the 5'- and 3'-terminal regions of the alpha1C gene. A possible existence of a new genomic homologue of the alpha1C subunit gene is discussed. PMID- 9748691 TI - 160Thr mutation in the rhodopsin gene associated with retinitis pigmentosa. AB - Mutations in the rhodopsin gene were studied in 23 unrelated Spanish patients with sporadic retinitis pigmentosa (RP). A codon 160 Thr C-->A transition was found in 4 of the 23 patients vs. none of the 159 controls (p < 0.001) suggesting that this mutation may be an informative marker in RP. PMID- 9748693 TI - New DNA polymorphisms define ethnically distinct haplotypes in the human transferrin receptor gene. AB - In a study of transferrin receptor (TFR) polymorphism in different ethnic groups using PCR and restriction cleavage we found a new Hin6I polymorphism in intron 7 and confirmed a tentative BanI polymorphism in exon 4 reported by Evans and Kemp [Gene 1997;199:123-131]. In all ethnic groups there was a complete and highly significant (p < 10(-10)) linkage disequilibrium where all BanI 1 alleles were linked to Hin6I 1 alleles. Furthermore in the European populations, but not in the Chinese, there was a close correlation between the three BanI-Hin6I haplotypes and the alleles of a previously described three-allelic RsaI polymorphism in the TFR gene studied by Southern blotting. There were distinct ethnic differences in TFR allele and haplotype frequencies. Thus the Saamis were significantly different from the other European ethnic groups, and the Lithuanians had a significantly increased frequency of the BanI 2-Hin6I 1 haplotype, suggesting that this marker may be informative in tracing prehistoric migrations and admixture by Baltic peoples. The new TFR polymorphisms and haplotypes may also be useful markers in studies of interactions with the transferrin and hemochromatosis genes, the genetic influence on body iron stores and disease associations. PMID- 9748694 TI - Adult-onset primary open angle glaucoma does not localize to chromosome 2cen-q13 in North American families. AB - Glaucoma is one of the leading causes of irreversible blindness in the world and is characterized by elevated intraocular pressure, optic nerve atrophy, and progressive visual field loss. Primary open angle glaucoma (POAG) is the most common subtype of glaucoma in the United States. Recently, Stoilova and coworkers [Genomics 1996;36:142-150] identified a locus for POAG on chromosome 2 (2cen-q13) in families primarily located in the United Kingdom. We examined families with POAG identified within the US for linkage to the 2cen-q13 locus. A total of 18 families with POAG were used in the analysis. Of 77 family members, 46 were classified as affected and 31 were either glaucoma suspects or considered normal. Eight highly polymorphic and informative markers flanking and distributed throughout the region were used. Parametric lod score analysis was performed using both a dominant and recessive low penetrance or 'affecteds-only' model. Multipoint affected sibpair exclusion mapping was also performed. Lod score (both models) and sibpair analysis excluded linkage of the POAG phenotype to the 2cen q13 region in these families. These data suggest that the chromosome 2cen-q13 locus does not explain a substantial amount of genetic variation in familial POAG. PMID- 9748695 TI - DNA analysis of the fragile X syndrome in an at risk pediatric population in croatia: simple clinical preselection criteria can considerably improve the cost effectiveness of fragile X screening studies. AB - Advances in understanding the molecular basis of the fragile X syndrome, the most common cause of inherited mental retardation, have elicited new prospects for population-based studies identifying affected individuals and fragile X families, thus contributing in prevention of the disease. In comparison with numerous fragile X screening studies where unselected groups of individuals with mental retardation, developmental delay, learning disability or autistic-like behaviour had been observed, we performed fragile X analysis on clinically preselected individuals. The group we studied consisted of 108 children with mental retardation of unknown cause or positive family history who had at least one physical and/or behavioural characteristic often observed among fragile X individuals. A relative high frequency of the fragile X positive cases (13% overall, 17.3% in males) was detected, suggesting that simple preselection criteria can considerably increase the proportion of fragile X-positive cases, and therefore, improve the cost-effectiveness of fragile X testing. Retrospective clinical analysis using a simplified six-item fragile X checklist confirmed that scoring criteria can be used to additionally preselect individuals at risk. Our results also indicate that this syndrome is underdiagnosed in Croatia and that a further effort must be made to detect unrecognised cases. PMID- 9748696 TI - Low-order polynomial trends of female-to-male map distance ratios along human chromosomes. AB - Recombination rates in humans tend to be sex specific. For a given map interval delimited by genetic markers, the difference between male and female recombination rates may be measured by the ratio, R, of female-to-male map distance. On average over all chromosomes, R is close to 2, but this ratio is region specific. The spatial variation of R can be captured by a low-order (linear, quadratic, etc.) trend across the length of the chromosome. Chromosome maps have been constructed that take into account such trends. Resulting map distances tend to be more accurate than when such trends are ignored. These maps may be obtained at URL ftp://linkage.rockefeller.edu/SexSpecMaps/. PMID- 9748697 TI - No evidence for an association between a variant of the mast cell chymase gene and atopic dermatitis based on case-control and haplotype-relative-risk analyses. AB - Atopic dermatitis (AD) is a chronic relapsing dermatitis which belongs to the group of atopy-related diseases as well as asthma and allergic rhinitis. As a probable genetic risk which may contribute to the organ specificity of AD, an association between AD and a genetic variant of the gene encoding mast cell chymase (MCC), which has chymotrypsin-like specificity and is abundant in skin mast cells, has been reported in a Japanese population. We tried to confirm the role of this polymorphism in the development of AD in a Japanese population. A case-control analysis using 100 AD patients and 101 controls did not show a significant difference in the frequency of the BB genotype between the patient and control groups (odds ratio 1. 12, p = 0.81). The haplotype relative-risk analysis using 69 patient-parents trios did not suggest an association (chi2 = 0.177, p = 0.92). Thus, we failed to confirm the association between the polymorphism in the MCC gene and AD in the Japanese population. PMID- 9748698 TI - Multi-locus nonparametric linkage analysis of complex trait loci with neural networks. AB - Complex traits are generally taken to be under the influence of multiple genes, which may interact with each other to confer susceptibility to disease. Statistical methods in current use for localizing such genes essentially work under single-gene models, either implicitly or explicitly. In genomic screens for complex disease genes, some of the marker loci must be in tight linkage with disease susceptibility genes. We developed a general multi-locus approach to identify sets of such marker loci. Our approach focuses on affected sib pair data and employs a nonparametric pattern recognition technique using artificial neural networks. This technique analyzes all markers simultaneously in order to detect patterns of locus interactions. When applied to previously published sib pair data on type I diabetes, our approach finds the same genes as in the published report in addition to some new loci. For a specific two-locus model of inheritance, the power of our approach is higher than that of the currently used analysis standard. PMID- 9748699 TI - A novel in frame deletion of codons 188-190 in the hMSH2 gene of a slovenian patient with hereditary non-polyposis colorectal cancer. PMID- 9748700 TI - Identification of a CA/TG repeat polymorphism proximal to the human DLX3 gene. PMID- 9748701 TI - Detection of the apoB-3500 mutation in a Russian family with coronary heart disease. PMID- 9748702 TI - Parental HLA genes, hormone level and offspring sex ratios: a reply to James' letter. PMID- 9748703 TI - Fomitellic acids, triterpenoid inhibitors of eukaryotic DNA polymerases from a basidiomycete, fomitella fraxinea PMID- 9748704 TI - Comparison of clinical efficacy of inhaled glucocorticoids. AB - Ratios of clinical efficacy of inhaled steroids are determined by re-evaluation of 29 clinical studies including 6538 patients. The method is based on the calculation of the areas under the curves (AUCs) describing the increase of morning peak flow over baseline during the period of treatment with inhaled glucocorticoids. Dividing the AUC by the applied dose results in a normalized AUC for a dose of 1 mg. Effects of treatments are compared for pairs of glucocorticoids tested in carefully matched groups using the normalized AUCs. Additionally, the effect of the drug delivery system is taken into account according to the literature because of its influence on the effective dose deposed in the lung. Thus, ratios of treatment success are determined: fluticasone propionate (FP, CAS 80474-14-2) versus budesonide (Bud, CAS 51333-22 3) 1:0.309, FP versus beclomethasone dipropionate (BDP, CAS 5534-09-8)1:0.561, FP versus flunisolide (CAS 3385-03-3) 1:0.339, FP versus triamcinolone acetonide (TAAC, CAS 76-25-5) 1:0.206, BDP versus Bud 1:0.609. Ratios for clinical efficacy correspond with the ratios of relative receptor affinities of these pairs of glucocorticoids. Besides the dominant influence of the receptor mediated effect the influence of local metabolisation in case of BDP is discussed as well as the effect of tissue binding in case of FP. PMID- 9748705 TI - Synthesis of a new homologous series of p-chlorophenyl alcohol amides, their anticonvulsant activity and their testing as potential GABAB receptor antagonists. AB - The anticonvulsant activity of a homologous series of p-chlorophenyl alcohol amides is described. The new compounds (+/-)-2-hydroxy-2-(4'-chlorophenyl) butyramide (2), (+/-)-3- hydroxy-3-(4'-chlorophenyl)pentanamide (4) and (+/-)-4 hydroxy-4-(4'-chlorophenyl)-hexanamide (6), were prepared and tested for their anticonvulsant activity. Compounds 2, 4, and 6 exhibited significant activity in seizures provoked by pentylenetetrazol. Chlorine in the para position of the phenyl ring increased both their potency at the peak effect and the duration of their anticonvulsant activity. The anticonvulsant activities of (+/-)-3-hydroxy-3 phenylpentanamide (3) and compound 4 were antagonized by DL-baclofen. This, and the protecting activity of 3 in the genetic-absence-epilepsy rats of the Strasbourg model suggest that the phenyl alcohol amides could be GABAB receptor antagonists. PMID- 9748706 TI - Effects of morphine, nalbuphine and pentazocine on gastric emptying of indigestible solids. AB - The effects of morphine (CAS 57-27-2), nalbuphine (CAS 20594-83-6) and pentazocine (CAS 359-83-1) on gastric emptying of indigestible solids were studied. In rats, which were fasted for 24 h, either morphine, nalbuphine, pentazocine or saline was injected intraperitoneally, and ten steel balls (1.0 mm in diameter) were inserted into the stomach. At 3 h, the number of balls which had passed into the small intestine was counted. If these drugs inhibited gastric emptying, naloxone was injected concurrently to study the mechanisms of the inhibitory effect. Morphine, nalbuphine and pentazocine significantly inhibited gastric emptying (ED50:0.041 [95% confidence interval: 0.0078-0.14] mg kg-1, 0.0012 [0.00037-0.0081] mg kg-1 and 0.81 [0.41-1.30] mg kg-1, respectively). Naloxone 0.3 mg kg-1 antagonized the inhibitory effect of both morphine 1.3 mg kg 1 (ED75) and nalbuphine 0.005 mg kg-1 (ED75). In contrast, naloxone 0.3 or 1.0 mg kg-1 did not antagonize the effect of pentazocine 1.9 mg kg-1 (ED75), but a higher dose of naloxone (3.0 mg kg-1) did so. Therefore, in the rat, morphine, nalbuphine and pentazocine inhibit gastric emptying of indigestible solids, and it is likely that morphine and nalbuphine inhibit the emptying through the same opioid receptor, whereas pentazocine does so through a different receptor interaction. PMID- 9748707 TI - Effect of the novel T-selective calcium channel antagonist mibefradil on kidney function in comparison with amlodipine. AB - In the present study, the renal effects of mibefradil (CAS 116666-63-8), a novel calcium channel antagonist which more selectively blocks T-type than L-type calcium channels, was tested by applying clearance techniques to anaesthetized rats. The effects of mibefradil on kidney function and on arterial blood pressure were compared with those of the long acting dihydropyridine-type calcium antagonist amlodipine (CAS 88150-42-9). The results show that, within a dosage range of 0.1 to 1.0 mg/kg i.v., mibefradil induced a dose-dependent decrease of arterial blood pressure. Kidney function was not significantly affected at a dose of 0.1 mg/kg. By increasing the dose to 0.3 mg/kg, mibefradil induced a significant increase in urine flow, renal sodium, chloride and potassium excretion. Also fractional sodium and chloride excretions were significantly enhanced at this dose. The diuretic and saluretic effects of mibefradil were accompanied by a significant increase in the glomerular filtration rate. At the highest dose of 1 mg/kg used, the blood pressure lowering effect of mibefradil was most pronounced and glomerular filtration rate rose only slightly and not significantly. At this dose, the enhancement of urine flow and urinary electrolyte excretion was smaller than at the dose of 0.3 mg/kg. The actions of mibefradil were qualitatively similar to those of the dihydropyridine derivate amlodipine which at a dose of 0.3 mg/kg produced nearly identical renal effects to mibefradil, but exerted stronger antihypertensive effects. This study demonstrates that mibefradil shares with amlidopine the property to induce, at appropriate doses, diuretic and saluretic effects with a concomitant increase in glomerular filtration rate. PMID- 9748708 TI - Changes in plasma and urinary norepinephrine following transdermal clonidine in spontaneously hypertensive rats. AB - To support a long-lasting antihypertensive effect of transdermal clonidine (CAS 4205-90-7), changes in plasma norepinephrine (NE) levels and urinary NE excretion as indices of the sympathetic nervous activities were investigated following transdermal and oral clonidine in conscious spontaneously hypertensive rats. Plasma NE levels were significantly reduced for 24 h during transdermal application of clonidine patch at 1.5 and 4.5 mg/kg on the back of each rat. Oral clonidine at 100 microgram/kg also lowered plasma NE levels. However, significant falls in the levels lasted only for 4 h after oral dosing. Urinary NE excretion was significantly decreased during both 4-8 and 8-24 h periods, and during an 8 24 h period following transdermal clonidine at 1.5 and 4.5 mg/kg, respectively. Significant decrease in urinary NE excretion was also produced during a 4-8 h period following oral clonidine at 100 micrograms/kg. Total urinary NE excretion during a 0-24 h period was dose-dependently reduced following transdermal clonidine, but was not altered following oral dosing. These findings suggest that the sympathoinhibitory effect of transdermal clonidine is more persistent than that of oral clonidine. Therefore, long-lasting antihypertensive effect of transdermal clonidine is closely associated with the sustained suppression of the sympathetic nervous activity. PMID- 9748709 TI - Pharmacokinetics of danaparoid sodium, dalteparin sodium and heparin determined by inhibitory effect on the activated coagulation factor X activity after single intravenous administration in rabbits. AB - The inhibitory effect on the activated coagulation factor X activity (anti-Xa activity) in plasma and urine of danaparoid sodium (DAS, CAS 9005-49-6) was compared with that of dalteparin sodium (DLS, CAS 9041-08-1) and heparin (CAS 9005-49-6) after single intravenous administration at a dose of 640 anti-Xa U/kg to male rabbits. The elimination of half-life of DAS was 9.90 h and was 6.0 times longer than that of DLS and 16.5 times longer than that of heparin. The area under the plasma concentration-time curve (AUC) of DAS was 47.13 +/- 14.55 anti Xa U.h/ml and was 2.4 times larger than that of DLS and 2.9 times larger than that of heparin. The urinary cumulative excretion of anti-Xa activity of DAS and DLS was 42.6 +/- 6.4% and 16.4 +/- 0.8% of dose, respectively, in 24 h after dosing, respectively. But the anti-Xa activity in urine was not detected at any sampling points after administration of heparin. DAS has a longer elimination half-life and a higher renal excretion of anti-Xa activity than that of DLS and heparin. Therefore, in comparison to DLS and heparin, it seems that the anticoagulant activity of DAS has a long duration. PMID- 9748710 TI - Bioavailability of escin after administration of two oral formulations containing aesculus extract. AB - In a steady-state cross-over study in 18 healthy volunteers, the relative bioavailability of beta-escin (CAS 11072-93-8) after oral administration of a new immediate release enteric-coated test formulation containing aesculus extract was evaluated in comparison with a prolonged-release reference preparation. The subject received the test and the reference preparation in randomised sequence for 7 days each with no washout period in between. The daily dose was 50 mg escin b.i.d. Blood samples for pharmacokinetic profiling were taken on the 7th treatment day of each period over a full 24-h cycle of two successive dosing intervals. For the determination of beta-escin serum concentrations, a highly specific radioimmunoassay (RIA) was used. Generally, escin serum concentrations were lower during the second dosing interval (night) than during the first interval, probably indicating a drug by food interaction. (The morning dose was given after overnight fasting whereas the evening dose was given between meals). Test and reference demonstrated bioequivalence with regard to the extent of absorption; for the AUC (0-24 h p.a.), the 90% confidence interval ranged from 84% to 114% (point estimate: 98%). The differences observed for rate parameters can be disregarded due to the generally slow elimination and the wide therapeutic concentration range of escin. PMID- 9748711 TI - Effects of inhaled KAA-276, a selective histamine H1 receptor antagonist, on antigen- and histamine-induced bronchoconstriction in animals. AB - The antiasthmatic profile of KAA-276 (1-[1-(4-fluorophenylmethyl)-1H benzimidazole-2-yl]-5-[2-[4-(2- carboxethyl) phenyl]ethyl]-1,5-diazacyclooctane sulfate, CAS 167264-26-8), a newly synthesized histamine H1 receptor antagonist, given by inhalation as an aerosol was investigated and compared with the profiles obtained using other routes of administration. When given by inhalation, or by intravenous or oral routes, KAA-276 inhibited antigen-induced bronchoconstriction in rats with ID50 (a dose to inhibit the antigen-induced response by 50%) values of 0.054%, 1 mg/kg, and 51.2 mg/kg, respectively. KAA-276 prevented the histamine induced wheal reaction in rats dose-dependently with ID50 values of 0.22% by inhalation, 0.18 mg/kg by the intravenous route, and 2.3 mg/kg by the oral route. To judge from these results, inhaled KAA-276, unlike intravenous or oral KAA-276, had no inhibitory effect on the histamine-induced wheal reaction at a dose (0.054%) that is effective against the antigen-induced airway asthmatic response. Inhaled KAA-276 suppressed antigen-induced bronchoconstriction in actively sensitized guinea pigs, and histamine-induced bronchoconstriction in monkeys. These results suggest that inhalation of KAA-276 would benefit patients with bronchial asthma without inducing unwanted systemic effects. PMID- 9748712 TI - High performance liquid chromatographic method for the determination of lobenzarit disodium in a sustained release tablet formulation. AB - A rapid and simple high performance liquid chromatographic method is described and validated for the determination of lobenzarit disodium (CAS 64808-48-6) in a sustained release tablet formulation. The calibration graph was linear over the range 20-105 micrograms/ml. The sensitivity (discriminator capacity) was 2.079 micrograms/ml. The coefficient of variations for repeatability and reproducibility were less than 1.60% and 1.30%, respectively. The accuracy of the method did not depend on lobenzarit concentration in tablets. The mean recovery was found to be 100.62%. The method was selective, even when degradation products were present. PMID- 9748713 TI - Comparison of the effects of various spasmolytic drugs on isolated human and porcine detrusor smooth muscle. AB - The spasmolytic activity of flavoxate (CAS 15301-69-6), anticholinergic agents oxybutynin (CAS 5633-20-5), and trospium chloride (CAS 10405-02-4), drugs commonly utilized in the therapy of hyperactive bladder, and phosphodiesterase (PDE) inhibitors papaverine (CAS 58-74-2) and vinpocetine (CAS 42971-09-5) on muscarinic contractions of detrusor smooth muscle strips isolated from human and porcine urinary bladder was studied in vitro using the organ bath technique. Trospium chloride was most effective in relaxing contractions elicited by muscarinic stimulation, while flavoxate was significantly less effective than all other drugs tested. The relaxing potency of oxybutynin was greater than those of PDE-inhibitors papaverine and vinpocetine but 3,000 fold less significant than those of trospium chloride. The effects of the individual drugs on muscarinic tension of both human and porcine detrusor muscle strips were nearly equal. The present results suggest that the pig might be an appropriate animal model for the study of effects of spasmolytic substances on the contractility of urinary bladder smooth muscle in vitro. PMID- 9748714 TI - Amino derivatives of phenyl alkyl thiophene as inhibitors of bone resorption. Structure-activity relationship. AB - Metabolism of arachidonic acid through the 5-lipoxygenase (LO) pathway generates compounds that stimulate osteoclastic bone resorption; since LO metabolites might play a role in bone loss due to excessive resorption it was tried to develop a series of antiresorptive agents starting from an already known LO inhibitor. Of the 35 compounds synthesized, 11 strongly inhibited (10 mumol/l) retinoic acid induced bone resorption in cultured mouse calvariae; they were also tested for their effect on LO activity using rat peritoneal neutrophils, but no correlation could be drawn between inhibition of LO and bone resorption. Other pathways, still to be identified, must therefore be targeted by these compounds even though LO inhibition might contribute to their effects on bone. Two compounds selected for further studies were found active on parathyroid hormone-induced osteolysis, while they had no effect on basal resorption; they must, therefore, act at some key point in the process of activation of osteoclastic resorption. This series of compounds may represent a new way for the treatment of bone loss due to excessive resorption. PMID- 9748715 TI - Uptake into leishmania donovani promastigotes and antileishmanial action of an organometallic complex derived from pentamidine. AB - Iridium (Ir)-(COD)-pentamidine tetraphenylborate (CAS 225-75-4) was selected from a primary screening to be evaluated in vitro on three Leishmania (L.) strains comparatively to pentamidine used as reference compound. The IC50 values obtained from in vitro evaluation on promastigotes of L. major CRE 26, L. donovani DD8 and L. donovani LV9 were 3.9, 23.5, and 3.3 mumol/l for Ir-(COD)-pentamidine tetraphenylborate and 1.6, 7.7, and 3.9 mumol/l for pentamidine isethionate, respectively. Cytotoxicity on mouse peritoneal macrophages led to determine a chemotherapeutic index of 1.7 for Ir-(COD)-pentamidine tetraphenylborate and 4 for pentamidine. Considering L. donovani DD8, the uptake of iridium complex by the promastigotes was shown to be saturable with a Km value of 17.4 mumol/l and Vmax of 1.3 nmol/mg protein/2 h. After 2 and 4 h incubation of treated promastigotes in drug free medium the absence of Ir-complex efflux is in favour of intracellular drug binding. As a matter of fact iridium complex was shown to bind ribosomal subunits in vitro, with no effect on macromolecular biosynthesis. PMID- 9748716 TI - Effect of octreotide on the pathology of hepatic schistosomiasis. AB - In clinical practice, octreotide (CAS 83150-76-9) has its greatest impact in the management of bleeding varices. The present work is the first one which was undertaken to investigate the possible use of octreotide as an antifibrotic agent and to study its effect on hepatic vasculature in Schistosoma mansoni infection. The material of this investigation consisted of two groups of albino mice (A, B), subdivided each into normal control, infected control, subgroups treated with octreotide, praziquantel (CAS 55268-74-1), and a combination of octreotide and praziquantel. Groups A and B were sacrificed at the 8th week and the 18th week post infection, respectively. By analysis of the obtained results, octreotide induced a reduction of the portal pressure, the weight of the spleen and the liver, the liver egg load (number of eggs) granuloma size and cellularity, and of the degree of hepatic fibrosis quantified by serum N-terminal peptide of type III procollagen in serum, serum laminin and tissue collagen using a Picrosirius red dye assay. Moreover, the biochemical state of hepatocytes has been improved. The subgroups treated with octreotide in association with praziquantel revealed better results than the subgroups treated with praziquantel alone. These obtained data were analysed in terms of histological extent of liver fibrosis in sections stained with Masson trichrome and sirius red, hepatocytic and sinusoidal changes at an ultrastructural level and by immunohistochemical demarcation of endothelial cells of blood vessels through the determination of factor VIII-related antigen. The promising results detected in this study may encourage to further investigate the positive findings of this drug with the intention of its possible application on a clinical level. PMID- 9748717 TI - Pharmacokinetics of cefdinir and its transfer to dialysate in patients with chronic renal failure undergoing continuous ambulatory peritoneal dialysis. AB - Cefdinir (CAS 91832-40-5) was administered orally as a 100-mg capsule (Cefzon) to a total of 12 patients with chronic renal failure undergoing continuous ambulatory peritoneal dialysis (CAPD) to investigate changes in the serum concentrations, excretion rate into the dialysate and serum-protein binding of cefdinir. Cmax values were 1.64-4.34 micrograms/ml, t1/2 values were 10.8-21.9 h., and AUC values were 31.1-73.1 micrograms.h/ml (0-30 h) in four patients given a single oral dose of 100 mg of cefdinir as a capsule. About 1 microgram/ml of cefdinir had still remained in the blood of all the patients 24 h after administration. The serum concentrations of cefdinir were dose-dependent in four patients of each group who were given an oral daily dose of 100 mg for 3 to 8 days and 200 mg (2 capsules) for 4 to 14 consecutive days. No marked change in laboratory test values or clinical symptoms before and after administration were observed in these dose regimes. Protein levels of 5.17-5.71 g/day were eliminated from the peritoneal dialysate and urine. Cefdinir inhibited 90 to 100% of the clinical isolates such as Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli and other enteric bacteria causing catheter infection and peritonitis, and its antibacterial activity was stronger than that of amoxicillin (CAS 26787-78-0) or cefaclor (CAS 53944-73-3) against these clinical isolates. PMID- 9748719 TI - 20th Congress of the European Society of Parenteral and Enteral Nutrition. Nice, France, 16-19 September 1998. Abstracts. PMID- 9748718 TI - Appropriate mammalian expression systems for biopharmaceuticals. AB - Process development for biopharmaceuticals is dictated by product quality, drug safety and economy of the manufacturing process. Not surprisingly, these factors also play a key role in the evaluation of mammalian cell expression systems to be used in the production of pharmacologically active glycoproteins. To date, the most prominent candidates for efficient expression of glycoproteins are mammalian cell lines such as mouse fibroblast cells (C 127-BPV), Chinese hamster ovary cells (CHO-DHFR, CHO-NEOSPLA, CHO-GS), mouse myeloma cells (NSO-GS) as well as transgenic animals carrying c-DNA or genomic DNA which codes for the protein of interest. The expression titer in the case of glycoproteins is mainly determined by the promoter construct, the site of integration into the chromosome, the copy number and the type of protein in question. Based on expression titer, CHO NEOSPLA and NSO-GS expression systems are most effective in the production of monoclonal antibodies and, to a lesser extent, of recombinant DNA derived proteins. However, based on overall product yield, expression of recombinant DNA derived proteins in transgenic animals is by far the most promising system. Therefore, for proteins required in large quantities, transgenic expression systems offer an attractive choice. However, cost of goods for products for which the dosage or the overall annual quantities are low, is dominated by downstream processing, filling, lyophilization and packaging and not by the fermentation process. Such proteins are preferentially produced by classical mammalian cell culture systems. Concerns which have to be addressed with respect to drug safety in the transgenic animal approach are the size of the herd, genetic stability from animal to animal, variation in productivity and in impurity profiles during lactation periods, microbial, viral, mycoplasma and prion contaminants, the dependence on health status and the life span of the animal. In a number of cases glycosylation of the protein is relevant for the prevention of immunogenicity of the protein, the pharmacological activity, the pharmacokinetic profile, solubility and stability against proteolysis. The glycosylation pattern, depending on protein structure, is influenced by the enzymatic system of the host cell as well as by fermentation conditions. Therefore, selection of host cells and culture conditions must take into account the requirement for a specific and stable glycosylation pattern. For the assessment of glycovariants, a number of protein analytical methods such as peptide mapping, isoelectric focusing, oligosaccharide mapping, MALDI-TOF (matrix assisted laser desorption mass spectrometry-time of flight), capillary electrophoresis and specific potency assays are available. In our experiments, glycosylation of proteins expressed in CHO cells was demonstrated to be very stable. Only extreme process times, cultivation methods and ammonium ion concentrations had an influence on the glycosylation profile. Among the three products investigated--tissue plasminogen activator (t-PA), interferon omega and soluble intercellular adhesion molecule (s ICAM)--t-PA expressed the most stable glycosylation pattern. Only at extreme ammonium concentrations an increase of mannose-5 structures was observed, whereas biantennary complex structures were reduced. On the other hand, interferon omega and s-ICAM showed greater susceptibility to increased ammonium concentrations and to adherent cultivation. Such conditions induced quantitative changes to the glycosylation pattern favoring the appearance of higher branched structures. Short cultivation times resulted in more heterogenous oligosaccharide structures. Since the glycosylation of the three proteins is different in the same host cell, the amino acid sequence of the protein apparently influences the glycosylation pattern and its sensitivity to culture conditions. In NSO-mouse myeloma cells, production of s-ICAM is two times as high as in CHO cells PMID- 9748720 TI - Joint Congress of the European Association of Nuclear Medicine and the World Federation of Nuclear Medicine and Biology. 30 August-4 September 1998, Berlin, Germany. Abstracts. PMID- 9748721 TI - Dietary influence on the urinary excretion of polyamines. AB - The amino groups of amino acids, the constituents of proteins, are catabolized in the urea cycle. One intermediate of this cycle, ornithine, is a precursor molecule of polyamines. The influence of dietary protein intake on the production and excretion of polyamines in the urine is yet unclear. The aim of this study was to investigate the excretion of polyamines in the urine following three days of creatine-free, creatine-free and low-polyamine diet in four persons. On the fourth day they were loaded with creatine-free, creatinine-free and low-polyamine high-protein diet (80 g/70 kg body weight). High-protein diet resulted in no increase of urinary polyamine excretion. The low-polyamine diet caused a non significant decrease in urinary polyamine excretion (by 15%). PMID- 9748723 TI - [Cerebrospinal fluid proteins in the diagnosis of disorders of the blood cerebrospinal fluid barrier in central nervous system diseases]. AB - BACKGROUND: Many neurological diseases are connected with the dysfunction of blood-CSF barrier. The quantitative determination of CSF proteins has already been used in the diagnosis of barrier impairments and inflammatory diseases of the central nervous system. PATIENTS: Serum and CSF, totaling 264 samples, were obtained from 15 controls and 117 patients with various diseases of the nervous system. Laurell's electroimmunoassay was used for estimation of albumin and IgG levels in serum and CSF. CSF-protein profile was evaluated according to Reiber's graph for the evaluation of the CSF-protein profile. RESULTS: The graph for the protein profile can be divided into 5 functionally different parts (1--normal range, 2, 3, 4--different types of barrier dysfunctions and 5--local humoral response in CNS without any barrier impairment). There was a good correlation of CSF-protein profiles and neurological diseases in our group of patients. CONCLUSIONS: According to our results, Reiber's graph was helpful for the diagnosis of blood-CSF-barrier dysfunctions. The graph has the following advantages: a) possibility of simultaneous assessment of the functional state of blood-CSF-barrier and the inflammatory response of the CNS, b)sensitivity for the determination of pathological local IgG-production in CNS and c) minimal number of protein assays necessary. PMID- 9748722 TI - [The effect of uric acid, creatine phosphate and carnitine on lipid peroxidation in cerebral cortex and myocardium homogenates]. AB - BACKGROUND: Uric acid as the product of purine nucleotide degradation is an integrate component of blood plasma. This metabolite is considered to be one of the important naturally occurring antioxidants building up the antioxidation system of the organism. Creatine phosphate and carnitine are important substances participating in energy metabolism of the cells. Energy production is closely related to the level of reduction systems and thus also to the antiradical ability of the cell. By this mechanism could creatine phosphate and carnitine improve the antioxidative capacity of the cell. METHODS: In homogenates of rat brain cortex and myocardium was the production of oxygen radicals stimulated by mixture of Fe2+ ions and ascorbate. Oxygen radicals may induce lipid peroxidation by the means of the reaction with lipid structures. We tried to inhibit the process of lipid peroxidation by addition of uric acid, creatine phosphate and carnitine into the incubation medium. Intensity of lipoperoxidation was measured by detection of substances giving positive reaction with thiobarbituric acid (TBA) in homogenates of brain cortex and myocardium. RESULTS: Uric acid in concentrations of 1 and 0.5 mmol.l-1 markedly inhibits the production of compounds reacting with TBA. This effect was not found in 0.05 mmol.l-1 concentration. Creatine phosphate and carnitine in 1 mmol.l-1 concentrations also decreased the value of lipid peroxides in homogenates of brain cortex, but their effect was lower than the effect of uric acid. This effect was not seen in myocardium homogenates. PMID- 9748724 TI - [Toxicologic importance of iron and copper atoms and their relation to reactive oxygen metabolites]. AB - Free radicals are, on the one hand, necessary for the physiological function of some systems but, on the other had, they play an important pathologic role. The formation of free radicals can be the result of various conditions and can initiate various diseases. Their formation may also be a consequence of certain pathological state of the organism. In this way generated free radicals can cause in the secondary to the damage organism. The metabolism of free radicals is significantly influenced by transition metals. These metals are present in the organism at given oxidative state chelated in the proteins. In this form the metalloproteins to have unique catalytical and redox properties. The transition metals are a part of an active centre of many enzymes. Iron and copper are the predominant transition metals in human organism. These metals are vitally important, but if present in high concentration, in unsuitable oxidative state and at improper site, they can catalyse the formation of highly toxic reactive metabolities of oxygen, for example hydroxyl radicals. The toxic damage may be direct if the metals are present in high oxidative state Fe(IV) or Fe(V). These "ferryl" compounds are strong prooxidants. The organism maintains the iron metabolism in equilibrium. If the iron plasma concentration reaches 40 micro mol/L, it is a sign, that iron is released from physiological protein structures and forms so called "catalytically active iron". In this form iron can be involved in Fenton reaction in which hydroxyl radical is formed. The article discussed the toxic effect of "catalytically" active iron at a given oxidative state with its influence on some diseases. PMID- 9748725 TI - [Changes in phospholipids after repeated induced sublethal spinal cord ischemia in rabbits]. AB - BACKGROUND: Degradation of membrane bound phospholipids in CNS during ischaemia begins with extreme rapidity. Sublethal ischaemia influences ischaemic tolerance in the affected neurons and is stressful enough to induce neuronal changes such as postischaemic hypoperfusion, transient suppression of protein synthesis and induction of stress (HSP) proteins. It seems, that the nature of factors responsible for ischaemic tolerance may involve the activation of multiple different systems. MAIN PURPOSE: The aim of this study was to investigate the changes of phospholipids in gray matter regions of spinal cord following sublethal ischaemia repeated in long intervals of reperfusion. METHODS: Male rabbits, weight range 2.5-3.5 kg were used in the experiment. They were divided in following groups : 1. control animals; 2. animals subjected to 25 min ischaemia; 3. animals subjected to 25 min ischaemia and 3 h of reperfusion; 4. animals subjected to sublethal (8-8-9 min) ischaemia repeated in long-lasting (8 8-24 h) intervals of reperfusion. Phospholipids were separated by thin layer chromatography, lipidic phosphorus was assessed spectrophotometrically. RESULTS: Sublethal ischaemia repeated in long-lasting intervals of reperfusion increased the concentration of phospholipids to control levels in all gray matter regions. The resynthesis in the dorsal horns, of PC and PE in the ventral horns and of PC in the intermediate zone. CONCLUSIONS: An excessive renewal of phospholipids after sublethal ischaemia repeated in longer intervals of reperfusion was most pronounced in the eh dorsal horns of the spinal cord and can be the result of many defensive cellular mechanisms. PMID- 9748726 TI - [Inhibition of lipid peroxidation in erythrocytes by aminoguanidine, resorcylidine aminoguanidine and their oxygen and sulfur analogs in diabetic rats]. AB - This study examined the effect of aminoguanidine (AG) and its structural analogs semicarbazide (SK) and thiosemicarbazide (TSK), as well as their condensation products with 2,4-dihydroxybenzaldehyde-resorcylidene aminoguanidine (RAG), resorcylidene thiosemicarbazone (RTSKon), and resorcylidene semicarbazone (RSKon) on erythrocyte lipid peroxidation in rats with diabetes mellitus induced by hydrogen peroxide. All of the tested compounds at concentrations 1 mmol.l-1 in incubation mixture significantly inhibited the formation of malondialdehyde (MDA), an end product of lipid peroxidation, as assessed by its thiobarbituric acid reactivity. AG and RAG were the most effective inhibitors of lipid peroxidation 90%). It was also found, that RSKon and RTSKon were more potent inhibitors of lipid peroxidation (70 and 80%) compared to Sk and TSK (50%). We suppose that this increase of inhibitory effect by compounds with resorcylidene group may be due to the formation of quinone structure. PMID- 9748727 TI - [Importance of advanced glycation end products--AGE products]. AB - Recent experimental findings suggest that free oxygen radicals and AGEs may be significantly involved in the onset and development of chronic diabetic complications and Alzheimer's disease. The presented review summarizes knowledge on structure and rise of these products in vitro and in vivo and the chemical and biological properties of advanced glycation endproducts are discussed. Strategy of influencing development and prevention of diabetic complications in the near future involves a potentially promising antiglycation therapy and supplementation by antioxidants. PMID- 9748728 TI - [Neurochemical changes associated with ischemic-reperfusion injury of the CNS]. AB - Ischaemia-reperfusion injury of the central nervous system is the third leading cause of death in the European countries. Since nerve cells are exclusively dependent on glucose oxidation, reduction of the glucose and oxygen delivery to the neurons affect all cellular metabolic pathways. Decrease of ATP production and its concentration as well as activation of ionic gradients, seems to be a primary process. Several degrading cellular enzymes, as lipases, phospholipases, proteases and endonucleases, are activated, in contrast to the severe inhibition of protein and phospholipid synthesis. Signal transduction pathways, playing a role in the regulation of cellular metabolism, are depressed due to the energy deprivation. After certain time of ischaemia, reperfusion does not lead to restoration of normal cell metabolism and intracellular situation could be even worsen, depending on the neuron type and duration of ischaemic period. Inhibition of glucose oxidation, decrease of ATP concentration, inhibition of proteosynthesis and signal transduction disturbances were observed during the period of reperfusion. Despite of proteosynthesis inhibition, several proteins, especially transcription and growth factors are overproduced. The loss of intracellular homeostasis, as the consequence of the above mentioned processes, could lead to neuronal death. PMID- 9748729 TI - [The effect of ischemia and ischemia-reperfusion on ion transport systems]. AB - Interruption of cerebral blood flow leads to dissipation of ionic gradients as the consequence of ionic channel overstimulation and ionic pump failure. The aim of this work was to study the possible effects of ischaemia and ischaemia followed by reperfusion on biochemical properties of endoplasmic calcium pump and synaptosomal sodium pump and sodium/calcium exchanger. The results presented in this study showed that 15 minute ischaemia led to the inhibition of all three ionic transport systems, however in different degrees. 60 minute reperfusion following 15 minute ischaemia led to partial recovery of calcium pump and sodium/calcium exchanger. The activity of sodium pump was still significantly depressed. Ischaemia and ischemia followed by reperfusion did not affect kinetic parameters of calcium pump. On the other side, both ischaemia and ischaemia reperfusion led to an increase of sodium pump affinity to ATP and a decrease of the enzyme affinity to potassium. The possible causes of the changes, as the alteration of membrane structure or altered enzymes phosphorylation are discussed in the study. In addition to the inhibitory effect of ischaemia-reperfusion injury, intracellular water accumulation, as the possible consequence of altered ion homeostasis, is documented by nuclear magnetic resonance (imaging). PMID- 9748730 TI - [Human cloning. (Do we have too much courage or not enough?) New perspectives in medicine]. AB - Cloning mammals including man has become a real possibility of these years. The result obtained in sheep and cattle demonstrated that technical problems have been solved and now other disciplines--philosophy, ethics, law etc are confronted with this challenge. On one side we have to look for all risks and adopt adequate responsibility, on the other side to consider medical benefits by solving problems with reproduction, organ transplantation and inherited diseases. As the last but not least we have to establish formal rules and to introduce the conception into all disciplines involved, in the first place into genetics. Trying to contribute to the adaptation of formal genetics to the new situation we propose to link the donor and the recipient of the nucleus with a triple line in pedigree schemes. PMID- 9748731 TI - [The modern approach to diagnosis and treatment of pollen allergy]. AB - Pollinosis is a seasonal multisytemic allergic disease the incidence of which is rising in the second half of the 20th century worldwide as well and also in the Czech Republic. The most frequent clinical manifestation of pollinosis is allergic rhinoconjunctivitis, less frequent are asthmatic complaints, there are however also other manifestations depending on the affected target organ. The diagnosis is based on a carefully recorded case-history, more accurate information and confirmation of the diagnosis is obtained from skin tests, in some cases assessment of specific IgE antibodies. The most important therapeutic procedure is prevention or at least restricted contact with pollen. Causal therapy involves properly indicated and performed specific immunotherapy by high quality standardized allergens. Pharmacotherapy of pollinosis, includes general treatment, in particular using second generation antihistaminic substances, and local treatment of nasal symptoms of pollinosis, the use of nasal steroids mastocyte stabilizers and topical antihistamines. In the treatment of ocular symptoms dominates the use of mastocyte stabilizers and topical antihistamines. For rapid control of acute nasal and ocular manifestations preparations containing decongestants are useful. An integral part of treatment is always systematic education of patients. PMID- 9748732 TI - [Immunomodulation in the treatment of atopic dermatitis]. AB - In the submitted review the authors present information on possibilities of immunomodulating treatment of atopic dermatitis. Immunosuppressive treatment comprises systemic corticosteroid administration (possibly combined with cytostatic immunosuppressive preparations), cyclosporin A and phototherapy. Immunopotentiating treatment of AD makes use mainly of modulating preparations with the nature of physiological substances close to the immune system, substances obtained from natural biological sources (thymic hormones) or in recent years more frequently substances of recombinant origin (interferons, interleukins). The use of immunopotentiating drugs of chemical origin (synthetic substances) is used less frequently in AD. The use of intravenous immunoglobulin preparations for severe forms of atopic dermatitis is only just starting. PMID- 9748733 TI - [Fatty acid composition of plasma phosphatidylcholine and levels of lipids and lipoproteins in hyperlipoproteinemia. II. Relation with B lipoproteins]. AB - BACKGROUND: Epidemiological trials provided evidence that the cholesterol concentration in lipoproteins B, i.e. VDL, IDL and LDL, correlate significantly with the incidence of ischaemic heart disease (IHD). The objective of the present study was to assess how the fatty acid composition in plasma phosphatidyl choline affects the total and LDL cholesterol, triglyceride and apolipoprotein B concentrations in subjects with primary hyperlipoproteinaemia and dyslipidaemia. METHODS AND RESULTS: In a group of 142 subjects with primary hyperlipoproteinaemia and dyslipidaemia the concentrations of plasma lipids, lipoproteins apolipoproteins and fatty acids in plasma phosphatidyl choline (PC) were assessed. The authors provided evidence by discriminant analysis where the dependent variables were the lower quintiles (Q1 + Q2) and the upper quintiles (Q4 + Q5) of concentrations of total cholesterol, triglycerides, LDL-cholesterol and apolipoprotein B and the independent variables were FA concentrations in plasma PC, that the total cholesterol concentration was inversely associated with the concentration of docosahexaenic acid (22:6n-3). The concentration of LDL cholesterol correlated inversely with the concentration of palmitoleic acid (16:1n-7). Triglyceridaemia was inversely associated with the linoleic acid concentration (18:2n-6). The concentration of apolipoprotein B correlated positively with myristic acid (14:0) and negatively with concentrations of oleic acid (18:1n-9) and linoleic acid (18:2n-6). CONCLUSIONS: The submitted results indicate that the fatty acid concentrations of PC in plasma are significantly and markedly correlated with concentrations of total cholesterol, triglycerides, LDL cholesterol and apolipoprotein B. It is possible that atherogenic lipoproteins may be favourably influenced not only by the amount of fat but also by a suitable fatty acid composition. PMID- 9748734 TI - [Helicobacter pylori CagA antigen antibodies]. AB - BACKGROUND: CagA antigen of Helicobacter pylori is highly immunogenic in humans. There is an increasing evidence that infection with CagA-positive strains is related to the development of peptic ulcer disease, atrophic gastritis, or gastric cancer. The aim of our study was to assess seropositivity to CagA in a group of 95 clinically symptomatic adults who underwent gastroduodenoscopy and to correlate results to their disease characteristics. METHODS AND RESULTS: Serum immunoglobulin G antibodies to CagA detected by ELISA kit (Helicobacter p120, Viva Diagnostika, Germany) were compared to standard IgG specific antibodies against a pool of H. pylori antigens Synelisa Pin plate, ELIAS, Germany). Immunoglobulin G antibodies to CagA were present in 5/31 (16%) serum samples from H. pylori negative persons and 10/28 (36%) serum samples from H. pylori positive patients without peptic ulcer disease compared with 8/11 (73%) H. pylori positive patients with peptic ulcer disease in the past, 11/13 (85%) H. pylori positive patients with duodenal ulcers or duodenitis and 4/5 (80%) H. pylori positive (1/7, 14% H. pylori negative) serum samples from patients with gastric resection for peptic ulcers in the past. Serum levels of antibodies to CagA in the groups of patients with peptic ulcer disease in the past, with present duodenal ulcers of duodenitis and in H. pylori infected patients with gastric resection were significantly higher then those of H. pylori infected patients without peptic ulcer disease (P < 0.05). On the other hand, there was no significant difference in the presence of the specific antibodies against at pool of H. pylori antigens between these four groups. CONCLUSIONS: These data suggest that serologic response to the CagA antigen is more prevalent in H. pylori positive persons with present or past peptic ulceration than among infected persons without peptic ulcer disease. The presence of antibodies to CagA in H. pylori positive persons may be useful for the identification of patients with higher risk or more severe disease. PMID- 9748735 TI - [Hypertension in diseases of the kidney. Pathogenesis and therapy]. AB - Renal disease is the cause of hypertension in about 5% of all hypertonics. Patients with renal hypertension are threatened by cardiovascular complications of hypertension even more frequently than patients with essential hypertension. Hypertension is moreover an important factor in the progression of renal insufficiency. In the pathogenesis of renal hypertension an important role is played by sodium and fluid retention and activation of the renin-angiotensin system. Progression of rental insufficiency can be retarded only by more strict control of hypertension than in patients with normal renal function. Optimal treatment is administration of angiotensin converting enzyme inhibitor which moreover in the majority of patients retards the progression of renal insufficiency more markedly than other antihypertensive drugs. PMID- 9748736 TI - [Paroxysmal nocturnal hemoglobinuria and its association with aplastic anemia]. AB - Paroxysmal nocturnal haemoglobinuria (PNH) is an acquired clonal disorder of haematopoiesis. Clinically it is characterized by intravascular haemolysis, venous thrombosis and often by bone marrow hypoplasia. Haemolysis and thrombosis develop as a consequence of deficiency of several proteins on the cell membrane of the affected clone of blood elements. This is caused by somatic mutations in the PIG-A gene, which encodes an enzyme involved in the biosynthesis of glycosylphosphatidylinositol (GPI) anchor. Spectrum of mutations in the PIG-A gene is different to that observed in other genes. The mutations are mainly small deletions and insertions causing frameshift; large deletions are rare. Recently, however, a 88 base pairs direct tandem repeat insertion has been reported in a patient with PNH developed on the background of aplastic anaemia (AA). The peculiar pattern of the PIG-A gene mutations and the finding that more than one mutated clone is commonly present in patients with PNH might suggest that some form of hypermutability, caused by decreased DNA stability, deficient repair or increased generation of mutagens, might underline PNH. As most mutations cause cell death, it would explain the hypoplastic nature of the disorder and its association with AA. Other models of pathogenesis of PNH are also discussed. PMID- 9748737 TI - [Selection criteria for treatment of severe hyperlipoproteinemias with LDL apheresis]. AB - LDL (low density lipoprotein) - apheresis has been established as an alternative management of severe hypercholesterolaemia after failure of conventional diet and drug therapy. General indication criteria for LDL-apheresis have yet been established. Indication guidelines in USA, Europe and japan are based on whether coronary heart disease is present and on the degree of lDL cholesterol elevation after treatment with diet and maximal drug therapy. It is reasonable to consider LDL apheresis therapy for: 1. patient with coronary heart disease and LDL cholesterol 4.9 mmol/l (190 mg/dl); 2. patients without coronary heart disease, but at high risk for disease (due to an LDL cholesterol above 6.4 mmol/l (250 mg/dl), a first-degree relative with premature coronary heart disease, and the presence of one or more additional risk factor. The therapeutical goal with present coronary heart disease is lDL cholesterol less than 3.4 mmol/l (130 mg/dl), with asymptomatic coronary heart less than 5.2 mmol/l (200 mg/dl). In addition, LDL apheresis is recommended for the management of all patients with homozygous familial hypercholesterolaemia due to the very high risk of coronary heart disease and the poor response to usual lipid-lowering treatments. In the end present two typical cases, treated by LDL-apheresis. PMID- 9748739 TI - [Erythrocyte membrane sodium transport in patients with type I diabetes mellitus (insulin-dependent diabetes mellitus)]. AB - BACKGROUND: Disorders in sodium metabolism such as an increased total body exchangeable sodium, were found in diabetic patients, although the underlying mechanisms were not clear. The aim of the study was to evaluate red blood cell sodium transport in patients with insulin dependent diabetes mellitus (IDDM) without diabetic nephropathy. METHODS AND RESULTS: Renal hemodynamics using the clearance of inulin and para-amino-hippuric acid during euglycemic clamp and red blood cell sodium transport were examined in 13 IDDM patients without microalbuminuria and in 12 weight-, age- and sex-matched healthy controls. Despite normal renal hemodynamics and intracellular sodium concentrations (6.57 +/- 1.45 vs 5.95 +/- 0.60 mmol/l), IDDM patients had lowered clearance of sodium (2.22 +/_ 1.11 vs 3.24 +/- 1.32 ml/min; p < 0.01) and increased activity of natrium-lithium countertransport compared to C (0.76 +/- 0.50 vs 0.31 +/- 0.22 mmol.l-1 .h-1; p < 0.01). No significant differences between IDDM and C were found in Na+-K+ pump (7.95 +/- 1.95 vs 6.9 +/- 0.99 mmol.l-1 .h-1), in Na+-K+ cotransport (0.68 +/- 0.82 vs 0.82 +/- 0.71 mmol.l-1 .h-1) and in passive Na+ permeability (0.11 +/- 0.05 vs 0.09 +/- 0.02 mmoll.l-1 .h-1). CONCLUSIONS: IDDM patients without signs of diabetic nephropathy have shown changes in sodium lithium countertransport which could play a role in the pathogenesis of diabetic nephropathy and hypertension in the course of the disease. PMID- 9748738 TI - [Mitochondrial myopathy, deafness and type 2 diabetes mellitus with tRNALeu(UUR) point mutation in mitochondrial DNA]. AB - BACKGROUND: A heteroplasmic A3243G point mutation in tRNALeu(UUR) gene of mitochondrial DNA (mtDNA) is found in patients with MELAS syndrome (Mitochondrial Encephalomyopathy, Lactic Acidosis and Stroke-like episodes), less frequently in patients with other dominating clinical features, such as deafness, diabetes mellitus type 2, hypertrophic cardiomyopathy, renal problems or inborn development defects. Present report describes histochemical, enzymatic and molecular biology studies of the family with clinical variant of meals syndrome. METHODS AND RESULTS: A 45-year-old woman with progressive muscle weakness, external ophtalmoplegia, perceptive deafness, ischemic heart disease, diabetes mellitus type 2 and hyperlactacidemia was metabolically investigated because the multiorgan problems indicated mitochondrial origin of the disease. Muscle biopsy revealed pronounced myopathic changes, ragged red fibers and decreased activity of respiratory chain enzymes - succinate cytochrome c reductase (< 5% control) and cytochrome c oxidase (< 10% control). Restriction fragment analysis of mtDNA from muscle, blood and hair follicles detected heteroplasmic A -> G mutation in the position 3243 of the tRNALeu(UUR) gene, which was more pronounced in muscle (28% of total mtDNA) than in blood (12%) or in hair follicles (10%). No mutation was found in blood and hair follicles of patient's mother and daughter. CONCLUSIONS: Diagnostics of mitochondrial diseases require close collaboration of clinicians with specialised laboratories. Treatment of mitochondrial disorders is only symptomatic, however, early diagnosis of the molecular defect is important for genetic counselling. PMID- 9748740 TI - [Hypertension in kidney failure and its treatment]. AB - Hypertension is found in more than 80% of patients with terminal chronic renal failure. Its etiology is multifactorial, however sodium and fluid retention are decisive. The basic treatment involves attainment and maintenance of optimal "dry weight". Antihypertensive treatment is necessary in only roughly 30% patients. Hypertension after renal transplantation is most frequently associated with immunosuppressive treatment (steroids, cyclosporin), however, it is important to consider also the role played by chronic graft rejection and rule out stenosis of the graft artery. Severe hypertension complicates also some types of acute renal failure, while malignant hypertension may be its cause. PMID- 9748741 TI - [Case reports of disturbances of effective osmolality regulation in disorders of the central nervous system]. AB - The topic of the effective osmolality disorder in the patients with the central nervous system injury is documented with the four selected cases from the case collection monitored in the Department of Anesthesia and Intensive Care. The syndroms were diagnosed using the computer programme evaluating renal functions in relation to homeostasis. Authors present two case reports of the Inappropriate ADH Secretion Syndrome (IADHS), one of the Cerebral Salt Wasting Syndrome (CSWS) and one of Diabetes Insipidus (DI). The cases were recorded with the aid of the computer programme working with the 13 routinely monitored values and 12 output parameters. Two case studies were retrospective. Mentioned four cases are the example of the effective use of the currently monitor values in the intensive care setting. According to the incidence of the effective osmolality disorders, inputing quickness and programme simplicity authors reccomend the routine use of the programme in the central nervous system damaged patients. PMID- 9748742 TI - [Use of cyclosporin A in hematology]. AB - We present an overview of clinical conditions where cyclosporin A (CyA) could be used in the treatment of hematological diseases. CyA is an effective immunosuppressive agent. It has long been successfully used in organ transplants and bone marrow transplants. The strong immunosuppressive effect of CyA makes this agent useful in various autoimmune diseases. The indication of CyA in certain hematological diseases has recently been steadily expanding. Based on the influence of CyA on cell immunity, the creation of cytokines and subsequently on humoral immunity we try to elucidate or justify its use in specific hematological diseases with regards to their pathophysiology. The common denominator to the diseases discussed below is a certain form of autoaggression. We include for example idiopathic autoimmune hemolytic anemia (AIHA) idiopathic thrombocytopenic (ITP), Evans syndrome, pure red cell aplasia. In certain sense we can include here even aplastic anemia (AA) and myelodysplastic syndrome (MDS). Furthermore, it is possible to use CyA for its ingerence to the proliferation and activation of T-cells in some T-lymphoproliferations and in histiocytoses. In B lymphoproliferation the use of CyA will probably require further studies. We briefly note the possibility of influencing multiple drug resistance (MDR) with CyA. PMID- 9748743 TI - [Quality of life in patients with chronic obstructive pulmonary disease and bronchial asthma]. AB - Review of data in the literature on the quality of life and its assessment in chronic obstructive lung disease and in bronchial asthma. The authors mention the most frequently used types of questionnaires and results achieved when using them. General questionnaires include the Sickness Impact Profile or the short version of a very detailed questionnaire which has 36 questions with sub questions (SF-36 = Short Form-36). Specific questionnaires are focused on certain questions concerning different diseases. These questionnaires include SGRQ (St. George's Respiratory Questionnaire) which is used mainly in chronic obstructive lung disease. For this disease also the CRQ was developed (Chronic Respiratory Questionnaire) but its section on dyspnoea is not standardized. For evaluation of the quality of life of asthmatic patients several questionnaires exist, in particular for children. Several questions call for further standardization. The value of questionnaires is, however, beyond dount. They elucidate the situation which does not ensue even from detailed functional examination of the lungs or immunological examination. It appraises bodily and mental functions of man, restriction of his activity, the sensation of comfort and general evaluation of his health. Thus "classical" evaluation methods are extended by now non traditional ways of appraisal of diseases which have a high prevalence and thus also great impact in the population. PMID- 9748744 TI - [Accidents and children]. AB - BACKGROUND: The accident rate in the population, especially injuries to children and youths are in the forefront of attention in all the industrialized countries. The reason is that accidents are the main cause of all deaths in children of over one year of age and in adolescents and accident-linked morbidity in children is second only to diseases of the respiratory tract. The aim of the study was to obtain more exact data than those that were available in the Czech Republic previously and to create conditions for a draft of preventive measures that would bring rapid and effective results. METHODS AND RESULTS: The accident rate in children has been followed up in all age groups attending elementary school. The basic series was comprised of 192,832 children 6.0 to 15.11 years old. An accident was registered in 31 550 pupils. Registered were all accidents by the children in the course of one calendar year and had been treated by a physician. There has been found a statistically highly significant greater total accident rate in boys (18.7%) than in girls (13.9%) by the chi-square test P = 0.00001 at P < 0.05. The most frequent of injuries were fractures, dislocation of joints (55.2%) with girls being affected more frequently than boys. Most frequently the accidents took place at school (25.0%) and at home (21.5%). The greatest number of accidents took place during physical education and organized sports (29.4%) and during games unorganized sports (28.6%). A relatively large number of injuries was sustained in conflicts with a schoolmate (7.4%). CONCLUSION: Much greater attention should be devoted to the prevention of injuries in children, namely at home and at school, because it is not only a health problem that is in question, but also a social and economic one. Abroad, annual losses are estimated at thousands of millions of USD. PMID- 9748745 TI - [Regional chemotherapy in experimental cholangiocarcinoma in pigs]. AB - BACKGROUND: Primary cholangiocarcinoma (PCC) accounts for cca 9% of all carcinomas of the liver and biliary system. Its prognosis of always unfavourable and treatment is difficult. METHODS AND RESULTS: In 25 pigs strain BU PCC was induced chemically with the aim to test the action of regional chemotherapy on the tumor tissue. For treatment 5-fluorouracil, 5-fluoro-2-deoxyuridine, mitomycin C and doxorubicin was used either as monotherapy or in combinations. During chemotherapy a significant reduction of the tumour or fibrotization occurred, as compared with animals not receiving treatment. The median of survival was prolonged from 51.3 to 210.3 days. CONCLUSIONS: Regional chemotherapy is a significant constituent of oncological treatment of PCC. Complications of treatment are negligible and the rest for further treatment are encouraging. PMID- 9748746 TI - [Hypertension and its treatment in pregnancy]. AB - Hypertensive disorders complicate approximately 10% of all pregnancies, about half due to transient and essential hypertension and the rest due to preeclampsia that continues to be a major contributor to maternal and perinatal mortality. However, when hypertensive pregnancies are carefully monitored, the neonatal mortality is low. Therefore, identification of women destined to have preeclampsia is essential, and it is the major purpose of the new classification proposed by M. A. Brown and M. L. Buddle to better stratify those hypertensive pregnant women who are at high risk and need intensive inpatient management. Prophylactic low-dose aspirin appeared to prevent preeclampsia in some studies and to be reasonably safe; however, the effectiveness in reducing the incidence of severe preeclampsia and improving pregnancy outcome remains uncertain. The basic therapy for hypertension during pregnancy is now hydralazine, labetalol and methyldopa; for preeclampsia the cornerstone for treatment is magnesium sulphate and hydralazine intravenously, and small doses of diazoxide, if necessary. Diuretics have a dubious place in treatment of hypertension during pregnancy, and ACE-inhibitors are contraindicated. In severe preeclampsia and eclampsia, the only solution is delivery; better knowledge of etiology and pathogenetics is needed for effective and safe treatment of gestational hypertension, as well as careful blood pressure monitoring and adequate laboratory control. PMID- 9748747 TI - [Paleopathology and medicine]. PMID- 9748748 TI - [Epithelioid hemangioendothelioma of the liver. Accidental detection of a solitary focus resembling a hepatic cyst]. AB - The authors describe the case-history of a young woman with a solitary epithelioid haemangioendothelioma of the liver which on initial sonographic and CT examination reminded of a simple liver cyst. Aspiration biopsy under US control confirmed a solitary tumor evaluated under the influence of results of imaging methods, admitting the presence of a capsule, as a probable cholangiogenic adenoma developing into malignity. The patient was subjected to surgical extirpation of the tumor and the definite diagnosis of its vascular origin was assessed only during a detailed examination of the resected tissue. After an eight-month follow up the patient has no signs of relapse. PMID- 9748749 TI - [Relation between Helicobacter pylori infection and ischemic heart disease]. AB - Recently published studies discuss some of the extragastric associations of infection caused by Helicobacter pylori. The relation of coronary heart disease and Helicobacter pylori has been probably first reported by Mendall et al. and this topic still requires larger prospective studies to assess the risks and explanation of the exact mechanism of chronic bacterial infection in development of coronary heart disease in later life. In this overview we tried to summarise available information on this topic. PMID- 9748750 TI - [Bleeding gastroduodenal ulcers from the viewpoint of the surgeon]. AB - The article has discoursed of a diagnosis and a therapy of bleeding peptic ulcer. There has been used the group of the patients admitted at The Surgical Unit of The Faculty Hospital Motol in Prague during two and half years. Urgent endoscopy and endoscopic treatment of indicated findings have been performed as a common procedures. The surgical therapy has been necessary for the part of this patients. Operation's indications, types of the operations and their timing have been discussed. PMID- 9748751 TI - [Disturbances of effective osmolality regulation in disorders of the central nervous system and possible methods of monitoring]. AB - Authors deal in detail with the pathophysiology of the osmolal regulation. Besides hyperosmolality the secretion of antidiuretic hormone (ADH) in increased by hypovolemia and hypotension. Secretion of ADH is lowered in hypoosmolal states. All other mechanisms are preferebly volume regulating and they influence mainly retention and excretion of sodium. Authors discuss homeostatic effects of the renin-angiotensin-aldosteron system, effects of renal failure with prevailing glomerular or tubular function disorder, impact of diuretics, natriuretic peptides, digitalis-like hormone, urodilantin and influence of the other solutes. Disorders of the effective osmolality regulation are frequent in the cerebral affections that originate from trauma, vascular disease, inflammation or tumors. Hypoosmolality and hyponatremia are presented in two different conditions: Inappropriate Vasopressin Secretion Syndrome (IADHS) and Cerebral Salt Wasting Syndrome (CSWS). Quick differential diagnose is important because the treatment of both syndromes is essentially different. Typical cause of hypernatremia is central diabetes insipidus (DI). The group of available calculated renal function parameters is applied in the differential diagnosis of these syndromes. They are creatinin clearance, excretion fraction of water and sodium, electrolyte clearance and electrolyte free water clearance. Investigation of ADH and natriuretic peptide could be even misleading. Pathophysiologic consequence of the state given by inappropriate elevation of one hormone can be the elevation of the second one. PMID- 9748752 TI - [Attitudes toward condoms and experience with their use among sex education teachers]. AB - BACKGROUND: The sex education is influenced by knowledge and attitudes. The aim of presented study was to evaluate the attitudes toward condom among sex education teachers and their personal experience with its use. METHODS AND RESULTS: Attitudes toward condoms were measured by 10-items anonymous questionnaires according to Zverina and Lippert. Personal experience was subjectively assumed in 5-point scale. The samples consists of 230 sex education teachers, among them 193 (84%) women. The average age was 40.5 +/- 9.7 years, in the range from 19 to 60 years. 37 teachers (16%) assigned themselves as religious. Condom was evaluated as a good protection against HIV and other STD in almost all answers. Three fifths of teachers considered condom to be an excellent contraception. 48% teachers did not know if the majority of women dislike condom. One tenth respondents rejected the opinion that the men dislike condoms. One tenth of teachers has never experienced condom personally. 12 percent stated to use it regularly, among them are more represented men. The opinions about quality of condom and decreased sensitivity by condom differ believers from atheists and users from non-users. CONCLUSIONS: Attitudes toward condom are predominantly liberal among Czech sex education teachers. The personal experience with use is common but not regular. The religion has small influence toward the attitudes and use. PMID- 9748753 TI - L-arginine treatment in ischemia/reperfusion injury. AB - We tested whether treatment with exogenous L-arginine, the precursor of nitric oxide (NO), could protect the skeletal muscle from ischemia/reperfusion (I/R) injury. A rabbit hindlimb I/R model (2.5 h ischemia/2 h reperfusion) was used. Morphological changes were elucidated by morphometry. Plasma concentrations of malondialdehyde (pMDA), as well as L-arginine and L-citrulline content in the plasma and skeletal muscle were measured. I/R injury in the skeletal muscle was manifested by development of prominent interstitial edema (fraction of interfiber area was 26.23% vs 15.09% in sham operated control, p < .005) and severe microvascular constriction (capillary area was 11.41 microns2 vs 16.92 in control, p <.005). These changes were accompanied by increased pMDA levels, indicating a process of lipid peroxidation in the cell membranes. L-arginine treatment (4 mg/kg/min intravenously, for 1 h, infusion initiated 30 min before reperfusion) caused an intracellular accumulation of this amino acid in the SM. Intracellular concentrations of L-citrulline increased (201.0 mumol/dm3 after reperfusion vs 176.0 before ischemia onset, p < .005), suggesting stimulated endogenous NO synthesis. L-arginine treatment protected capillary constriction (capillary area was 17.64 microns2 vs 11.41 in the untreated animals, p < .0005) and reduced interstitial edema after reperfusion (fraction of interfiber area was 17.80% vs 26.23 in untreated animals, p < 0.005). The protective effect of L arginine treatment on I/R injury of SM may be related to its ability to prevent microvascular constriction and reduce permeability disorders by the stimulation of endogenous NO production. PMID- 9748754 TI - [Thromboelastography--an instrument for the researcher or clinician?]. AB - Thromboelastography is a method which is used experimentally since 1948. Since the end of the eighties it is experiencing a certain revival also in clinical medicine. The submitted case-record presents this technique as a very useful aid in the differential diagnosis of postoperative haemorrhagic conditions in cardiosurgery. Its application can facilitate aimed treatment of some typical disorders of haemocoagulation. PMID- 9748755 TI - [Recommendations for specialized care of alcohol and drug dependence and pathologic gambling]. PMID- 9748756 TI - [Effects of carvedilol, a sympatholytic, in experimental alloxan diabetes in laboratory rats]. AB - A possible effect of the sympatholytic carvedilol on alloxan-induced diabetes mellitus in the laboratory rat was examined in experiments. The animals were divided into a group treated with carvedilol in a single daily dose of 10mg/kg in 1 ml of diluting solution i.p and the control group which received only diluting solution in the pertinent amount. The values of malondialdehyde and glucose in the serum, diuresis and total losses of sugar in the urine within 24 hours were estimated and histopathological examination of the kidneys of the treated and control groups was performed. The results show an effect of the tested dose of the drug, primarily in the region the proximal renal tubule. PMID- 9748757 TI - [Anti-inflammatory activity of (cresoxyacetate)copper complexes]. AB - In groups of mononuclear aqua(cresoxyacetato)copper(II) complexes of the composition [Cu(ROCH2COO)2(H2O)n] (R= 2- and 3-methylphenyl, n = 2; 4 methylphenyl, n = 3) and binu- clear phenazone(o-, m-, p cresoxyacetato)copper(II) complexes of the composition [Cu2(ROCH2COO)4(phz)2] the anti-inflammatory activity was assayed in rat paw carrageenan-induced edema. The effects of the complexes were compared with each other and with those of the free acids. Salicylic acid and its cupric salt (tetrahydrate) were used as standards of comparison. All copounds were applied i.p. in a single dose of 50 mumol/kg body weight (calculated for the RCOO-fragment). In general, all Cu(II) complexes tested were clearly more effective (71-88%) than the corresponding free isomeric cresoxyacetic acids (47, 37, 45%), with the exception of diaquabis (o cresoxyacetato)copper(II) complex (43%). Copper(II) salicylate tetrahydrate (57%) and salicylic acid (41%) were less active, too. The observed activities of complexes are discussed in relation to their structures. PMID- 9748758 TI - [Antimicrobial activity of (N-salicylidene-DL-aspartate- and (N-salicylidene-L asparaginate)-copper complexes with pyrazole-type ligands]. AB - By a reaction of salicylaldehyde (Scl) with the corresponding amino acids and by the next complexation reaction of the formed Schiff bases with Cu2+ ions in an aqueous-alcoholic medium, aqua (N-salicylideneaminoalkanoato)copper(II) complex chelates of the composition Cu(Scl-DL-Asp(2-)) (H2O)2, Ip and Cu(Scl-L-Asn(2 )(H2O), In were prepared. The monodiazole complexes with pyrazole IIp and IIn (as monohydrate) as well as with 3,5-dimethylpyrazole IIIp a IIIn were prepared by replacing the molecule of H2O in the parent aquacomplexes with the diazoles under the same reaction conditions. Using a routine dilution micromethod, the antimicrobial activity of the prepared complexes and free diazoles was tested against Staphylococcus aureus, Escherichia coli and Candida albicans. Only a significant antistaphylococcus activity was found (highest for the complex IIn; MIC = 39 micrograms/cm3). All chelates (Ip,n-IIIp,n) were more effective (MIC = 39-156 micrograms/cm3) than both pyrazole (312 micrograms/cm3)and 3,5 dimethylpyrazole (625 micrograms/cm3) alone. The relationship between the coordination-chemical properties and the biological effects of the complexes studied is discussed. PMID- 9748759 TI - [Phospholipase A2: characteristics and function]. AB - Phospholipase A2 (PLA2) hydrolyses membrane phospholipids (PL) and it may release arachidonic acid (AA)--the precursor of eicosanoids--from the sn-2 position. PLA2 and metabolites of its catalytic activity participate in many processes in the organism: metabolism of lipids, inflammation and immune reactions, membrane and tissue reparation, proliferation, and others. PLA2 as also an important element in the signal transduction. In the present article, PLA2 is characterised, classified into several types, and its mechanism of action together with its possible role in the disease processes are described. The attention is aimed at two forms of PLA2: The secretory (PLA2) of the type II which is associated with the inflammation injury, and the cytosolic PLA2 which is the main catalyst in the liberation of AA and which participates in the signal transduction. Other forms of PLA2 has been also described. PMID- 9748760 TI - [Structure of the renin-angiotensin system and its significance in the body]. AB - Renin-angiotensin system (RAS) is a methabolic pathway producing octapeptid angiotensin II (AII) with vasoconstrictive effects, which is also involved in the regulation of water homeostasis and electrolyte balance in the organism. Genes encoding individual components of the RAS can be considered as "candidate genes" for some cardiovascular diseases e.g. hypertension or myocardial infarction. Besides the circulating RAS, also local tissue systems exist. Local RASs are involved in various physiological functions e.g. regulation of growth and proliferation, apoptosis or signal transmission where AII serves as the neurotransmitter. PMID- 9748761 TI - [Cross-regulation between muscarine and beta-adrenergic receptors]. AB - Muscarinic and beta-adrenergic receptors belongs to the family of G protein coupled receptors. Their activation brings about nearly antagonistic effects in most tissues. We demonstrate on the model of myocardial cells, that their regulation is mutually interconnected and that the cross-regulation could represent new level of homeostasis maintenance. PMID- 9748762 TI - [Medical cybernetics in Czechoslovakia--the first steps]. AB - During fifties there were at most few tens of persons in this country who believed in the future of computers and cybernetics. One group of such enthusiasts, headed by Antonin Svoboda, was working at a construction of the first Czech computer SAPO. The other group tried to analyse, anticipate, and prepare in advance various applications for the new systemic conceptions and for the information processing machines. Members of both groups met for discussions which opened prospects to the future and influenced many of other activities for a long time. At the early sixties, the Czechoslovak Cybernetic Society was established at the Czechoslovak Academy of Sciences and in 1962 the Main Problem Committee for the Medical Cybernetics was founded at the Department of Health. It coordinated majority of the research programmes in the medical cybernetics and informatics. In 1967-1969 the Committee prepared an extensive project of a medical information system (ZIS), but its accomplishment was finally blocked by the then authorities. However, interests for that topics kept growing and the new working places equipped with available computer technology were formed at the health and clinical centres. The first tentative lectures in medical cybernetics and biocybernetics at our faculty were introduced into the students curricula in the late sixties. Thematically, medical cybernetics subsequently differentiated into the medical informatics, simulations of biological and medical systems, and the biosignal analysis. The growing interest enabled to hold conferences since the middle of seventies, some of which were held periodically, sometimes with international participation. It is not possible in brevity to include the whole spectrum to those goal-directed activities nor to appraise adequately their future significance. PMID- 9748763 TI - [Evaluation of education at Danish medical schools]. PMID- 9748764 TI - [Personal experience with classification of yeast microorganisms. I. The combined biotyping method of Mencl and Otcenasek and typing using the "killer" phenomenon]. AB - Using a group of 150 isolates of Candida albicans, C. glabrata, C. krusei, C. tropicalis, C. parapsilosis a C. kefyr the differentiating capacity of two biotyping systems was tested-the combined method according to Mencl and Otcenasek, and typing using the so-called killer phenomenon. With the combined method comparable results with the original work of the authors were obtained. This applies to the number of biotypes as well as to the ratio of the dominant biotype. As regards the differentiating characteristics of different biotypes the two studies differed fundamentally. As to typing, using the "killer" phenomenon, its practical usefulness was tested, the differentiating capacity of the method was, however, very much influenced by the small number of available killer positive yeast strains. PMID- 9748765 TI - [Viral hepatitis B morbidity in the Trebisov District after introduction of vaccination]. AB - Hepatitis B morbidity in the Trebisov district (eastern Slovakia) exceeded several times the average morbidity in Slovakia in the first half of the eighties. High morbidity of children in the age groups 0-4 and 5-9, frequent extraparenteral transmission in families with sick members or with HBsAg carries (34.9%), led to the introduction of the regular vaccination of all children in their first year of life. Already during the first years of this vaccination, an important decrease of morbidity in vaccinated age groups was observed. PMID- 9748766 TI - [Giardiasis in clinical data]. AB - The authors hospitalized and treated in 1990-1994 at their Clinic 127 patients where giardiasis was the main or secondary diagnosis. They wanted to draw attention to the increasing prevalence of the disease, to clinical manifestations and problems associated with treatment. Clinicians should consider the possible presence of this disease in different gastrointestinal manifestations, chronic hepatitis, hepatopathies as well as in acute IgM anti-HAV, HBsAg and HCV negative hepatitis. PMID- 9748767 TI - [Urethritis of chlamydial origin]. AB - Using the direct immunofluorescence method with a Chlamyset of Orion Co. Finland for detection of Chlamydia trachomatis, the authors examined smears from male and female patients with manifestations of non-specific urethritis and cervicitis. In women also specimens from the cervix were examined. The IF examinations of smears from the urethra of 72 men were positive in 25, i.e. 34.7%. IF examinations of 70 women with manifestations of cervicitis and urethritis were positive in 17, i.e. 24.3% from the urethra and in 20 cases, i.e. 28.6% from the cervix. The positive findings of Chlamydia trachomatis indicate the importance of diagnosing urethritis of chlamydial origin and aimed treatment. PMID- 9748768 TI - [Tularemia, "volume" two]. AB - As a continuation of Libich's monograph (Tularemie. Prague, Avicenum 1981) the author presents findings assembled in experiments pertaining to postinfectious immunity on a model of intracellular infection with the microorganism Francisella tularensis. PMID- 9748769 TI - [Benzodiazepines are just a tool in the clinical work. Different opinions are both right and wrong]. PMID- 9748770 TI - [Increased risk of malignancy in primary sclerosing cholangitis. Liver transplantation is a therapeutic alternative if no cholangiocarcinoma occurs]. PMID- 9748771 TI - [Interpretation of the law on involuntary commitment by the National Board of Health and Welfare is against the intentions of the law!]. PMID- 9748772 TI - [Earmark the money for internship and residency!]. PMID- 9748773 TI - [Why nothing about the causes of violence?]. PMID- 9748774 TI - [Do we really want to decide about our death?]. PMID- 9748775 TI - [Pulmonary embolism remains an overlooked diagnosis. Proposed strategy in suspected cases]. AB - The problem of misdiagnosed pulmonary embolism (PE) is reviewed in the light of the introduction of new diagnostic methods such as spiral tomography (helical scanning). Despite new diagnostic methods, the frequency of misdiagnosed PE will not be reduced unless PE is suspected at the right juncture. PE should always be suspected in the presence of 'pulmonary syndromes' and venous thromboembolic risk factors. A strategy for the management of cases of suspected PE is proposed in the article: 1, recognition of the presence of cardiopulmonary disease, 2, determination of the clinical probability of PE; 3, confirmation (or exclusion) of PE; 4, determination of its severity and prognosis; and 5, choice of treatment. A two-part algorithm for use in patients with stable or unstable haemodynamics is also presented. PMID- 9748776 TI - [New therapeutic principle in severe hyperhidrosis. Botulinum toxin injections can replace cutting of nerves]. PMID- 9748777 TI - [Treatment of facial blushing with endoscopic thoracal sympathicotomy. 85 per cent of patients are satisfied, but there are adverse effects]. AB - Endoscopic transthoracic sympathicotomy, otherwise an established treatment for palmar hyperhidrosis, was used to treat patients troubled by facial blushing, one of the commonest symptoms of social phobia. The results were evaluated by means of a questionnaire answered by 90 per cent (219/244) of the patients, who rated their symptoms on a visual analogue scale (0-10) after a mean follow-up of eight months. According to the ratings, blushing was significantly reduced from a mean (+/- SEM) of 8.7 +/- 0.1 to 2.2 +/- 0.2 (p < 0.0001). Of the series as a whole, 85 per cent declared themselves satisfied with the outcome. PMID- 9748778 TI - [Demographic factors affect the occurrence of severe mental retardation. Is intermarriage a factor of significance also in Sweden?]. PMID- 9748779 TI - [New knowledge of heredity in inflammatory bowel disease. Specific gene mapping in a EU-project]. AB - Chronic inflammatory bowel diseases, ulcerative colitis and Crohn's disease, are steadily increasing in prevalence, and one half to one per cent of the Swedish population are currently estimated to be affected. The aetiology remains unknown, but is probably multifactorial. Both dietary, microbiological and immunological causes have been discussed. Clinical studies, including several Swedish studies, have also shown genetic factors to be crucially involved. Findings in sophisticated molecular biological studies suggest certain specific genes to be involved, and a current EU project in which Sweden is participating has been launched to map the mode of inheritance in detail. PMID- 9748780 TI - [Statins have other beneficial properties besides their cholesterol lowering effect]. AB - Statins, i.e. HMG-CoA (3-hydroxymethyl-glutaryl co-enzyme A) reductase inhibitors, are widely used in the treatment of hyperlipidaemia. Several large trials published during recent years have clearly shown treatment with statins to reduce coronary heart disease morbidity and mortality rates, the beneficial effects being manifest sooner than expected. Despite the smallness of changes in lumen diameter during statin treatment, as measured with coronary angiography, the decrease in clinical events has been impressive. The question has therefore arisen of whether statins have other beneficial effects in addition to their lipid-lowering property. Several studies have been made of the effects of statin treatment on such variables as endothelial function, cellular immunity, lipoprotein oxidation, rheological factors, and the stabilisation of atherosclerotic plaque. Although the various statins differ in structure, there is a lack of comparative studies. However, available data suggest the beneficial effects of statins on other variables to be probably dependent on their primary lipid-lowering property. PMID- 9748781 TI - [A five-year follow-up of 115 patients treated with anticoagulants. Bleeding complications may be underestimated]. AB - The prevalence of anticoagulant treatment in Sweden has increased in recent years. However, such treatment may be associated with a risk of serious complications. At 5-year follow-up of 115 primary care patients treated with anticoagulants in 1992, 39 were found to have died, eight of anticoagulant induced bleeding complications (six of intracranial haemorrhages, and two after profuse gastro-intestinal haemorrhages). In only two of these cases, had an adverse reaction been diagnosed. The fatal complication rate was estimated to be 2.1 per cent per treatment year. There were 17 major complications requiring hospitalisation, the estimated rate being 4.4 per cent per treatment year. Three patients died of thromboembolic episodes during anticoagulant treatment, and a further three after completed treatment. PMID- 9748782 TI - [Alendronate-induced severe esophagitis. A rare and severe reversible side-effect illustrated by three case reports]. AB - The bisphosphonate, alendronate sodium (e.g. Fosamax), a bone resorption inhibitor used to treat osteoporosis, has occasionally been reported to cause severe oesophagitis. The characteristic endoscopy findings include oesophageal ulceration with discoloured exudate, a narrowed lumen and denuded, haemorrhagic mucosa. The oesophagitis heals on discontinuation of alendronate medication, and the institution of gastric acid suppression treatment. The article consists in discussion of such adverse reactions, illustrated by three case reports. As many of the patients selected for alendronate treatment are elderly and handicapped, to minimise the risk of serious side-effects it is important not only to give detailed instructions regarding medication, but also to ensure that they are properly understood. As a history of swallowing problems was a feature of all three cases reported, caution is recommended before treating such patients with alendronate sodium. PMID- 9748783 TI - [Increased safety requirements concerning products of medical technology. New rules make CE labeling compulsory in most cases]. PMID- 9748784 TI - [Benzodiazepines are the right choice i certain anxiety disorders. A recent report from the WHO confirms the evidence-based experience]. PMID- 9748785 TI - [Bone mineral density in patients on long-term therapy with levothyroxine]. AB - The aim of the study was to determine the influence of replacement and suppressive thyroxine therapy on bone mineral density (BMD). 30 postmenopausal women; 19 on replacement therapy (dose 1.22 +/- 0.35 micrograms/kg; duration 11.4 +/- 7.2 years) and 11 on suppressive therapy (dose 1.45 +/- 0.71 micrograms/kg; duration 9.5 +/- 7.2 years). Controls were 60 healthy women matched for age and menopausal status. BMD at the lumbar spine (L2-L4), femoral neck, Ward's triangle and trochanter was measured by dual-energy absorptiometry. Forearm BMD at distal site was measured by single-photon absorptiometry. Mean thyroid hormone values and TSH were within normal limits, although the patients on suppressive therapy had significantly higher T3 (p < 0.05) than the patients on replacement therapy. BMD on each site was significantly lower in the replacement treated group than in controls. BMD in patients on suppressive therapy was lower, but not significantly, compared to controls. Thyroxine therapy could have an adverse effect on BMD. The magnitude of bone loss depends on the serum level of thyroid hormones and on the functional state of thyroid hormone receptor in bone tissue, as well. PMID- 9748786 TI - [Cytomegalovirus retinitis in patients with human immunodeficiency virus infection]. AB - Cytomegalovirus retinitis (CMVR) is a common opportunistic infection and a major cause of blindness in patients with AIDS. The aim of this study was to determine the frequency, clinical course and outcome of CMVR in patients treated at the University Hospital of Infectious Diseases "Dr. Fran Mihaljevic" in Zagreb in the period from January 1995 to April 1996. CMVR was diagnosed in 8 (27.5%) of 29 patients with AIDS. The median CD4 lymphocyte count in patients with CMVR was 44 per mm3, six patients had less than 50 per mm3. Five patients died during the study period, the mean survival being 5.5 months. CMVR was present in both eyes in 5 (62.5%) patients at the time of diagnosis. Blindness in both eyes developed in 3 (37.5%) patients. In order to recognize and promptly treat CMVR frequent ophthalmologic examinations should be performed in patients with advanced HIV disease. PMID- 9748787 TI - [Does an early heart attack in a parent indicate increased cardiovascular risk in the offspring?]. AB - In a 5-year period analyzed were cardiovascular risk factors in 55 patients suffering from premature coronary accidents (below the age of 45) and in their 97 offsprings, aged 3-26 years. In 50 of these children (51.6%), 22 girls and 28 boys, in addition to the obvious positive family history, detected were other cardiovascular risk factors. The most common among them was hypercholesterolemia (in 45.0%), followed by arterial hypertension (40.0%), obesity (32.0%) and smoking (24.0%). Majority of these offsprings (64.0%) had only one additional risk factor. The average blood pressure, relative weight and cholesterol levels were significantly higher in these very children than in all the coronary offsprings (p < 0.05), and even more so (p < 0.01) than in the matching control sample. PMID- 9748788 TI - [The Johanson-Blizzard syndrome]. AB - A 17 year and 10 month old boy with Johanson-Blizzard syndrome is presented as a case report for the first time. Diagnosis has been established on the basis of craniofacial abnormalities: microcephalia, parietal skin and bone defects, sparse hair with frontal up sweep, alae nasi hypoplasia, irregular dentition and nasolacrimal fistula, with mental insufficiency, partial exocrine pancreatic insufficiency and low birth-weight and length, hypotonia and failure to thrive in infancy. Congenital cataract and hiatus sacralis apertus are additional signs that have never been described in the literature concerning Johanson-Blizzard syndrome. PMID- 9748789 TI - [Comparison of the results of PCA knee joint endoprosthesis in patients with rheumatoid arthritis and gonarthrosis]. AB - Hundred and forty two porous-coated anatomic (PCA) total condylar arthroplasties were performed from 1985 to 1991 in hundred and twenty-four patients, ninety seven women and twenty-seven men. The diagnosis was osteoarthritis in 96 cases and rheumatoid arthritis in 46 cases. The mean follow-up was 88 months, range from 51 to 137 months. All operated patients were evaluated based on survivorship of the endoprosthesis with the cumulative survivorship method according to Kaplan Meier method. The end point was defined as endoprosthesis in situ. The overall cumulative survival rate for mean follow up time was 77%. The survival rate of the rheumatoid arthritis group was significantly higher (82.5%) than that of the osteoarthritis group (73.8%). 108 PCA arthroplastes were evaluated regarding Baltimore's score. The mean postoperative Baltimore's score was 68.7. In the rheumatoid arthritis group score was 74.8 and in the osteoarthritis group it was 65.9 but this difference is not statistically significant (p = 0.06). PMID- 9748790 TI - [Treatment of femoral neck fractures with bipolar hemiarthroplasty]. AB - The aim of the study was determine the effectiveness of treatment with bipolar prosthesis in elderly patients with the femoral neck fractures. Patients admitted from 1980 through 1994 were analysed. Over this period, a total of 152 hip joint bipolar prostheses were implanted. Postoperative complications occurred in 9.2% of cases (luxations of endoprostheses 2.63%, loosening of the femoral component 1.31%, infections 2.63%, intrahospital mortality 1.31%, fractures of the endoprosthetic stem 0.65% and fracture of the polyethylene cup 0.65%). Ten (6.5%) hips required revision surgery. Twenty eight patients were followed-up. Excellent results were reported in twenty-three patients (82.1%), good results in four (14.3%), and fair results in one patient (3.6%). There were no poor results. In summary, we feel that bipolar hemiarthroplasty is the method of choice in elderly patients with femoral neck fractures. PMID- 9748791 TI - [Use of methadone in the treatment of narcotic addiction]. AB - In spite of the 30-year experience in the use of methadone in heroin addicts, there is no consensus about the value of that method. In view of the opinion that resistance to methadone treatment is partly caused by ignorance, the aim of this paper is to improve the knowledge about the method. The paper offers explanation of: (1) metabolic basis of heroin addiction, (2) methadone pharmacology, (3) psychosocial aspects of the addiction, (4) methadone treatment procedure, (5) aims and (6) results of the treatment. A demand for designing the treatment program on the basis of expert knowledge rather than on political and moral judgments is stressed. Topics in program organization are described: indications, daily dosage and its administration, treatment duration, professional job-sharing and control of the treatment. A review of medical goals--individual-oriented and social-oriented goals of the treatment is made. "Therapeutic milieu", as well as treatment dose and treatment duration have been recognized as the main determinants of the outcome. Evaluation criteria and treatment have been elaborated as potential basis for further research. PMID- 9748792 TI - [Membrane metalloendopeptidase (CD10/CALLA): distribution, physiologic and pathophysiologic functions and its inhibitors]. AB - Membrane metalloendopeptidase EC 3.4.24.11 (Enkephalinase, neutral endopeptidase, NEP) is a cellular ectoenzyme, immunophenotypically identified as the leukocyte cluster of differentiation CD10 or CALLA (common acute lymphoblastic leukemia antigen). Immunological, biochemical and molecular biology techniques have identified tis cell membrane feature in various organs: brain, cardiovascular system, lung, placenta, kidney etc. The CD10 immunophenotype is a common feature of lymphoblasts in acute lymphoid leukemia not expressing the T- or B-markers. The enzymatic activity of CD10/NEP possibly influences normal lymphocyte ontogeny by proteolytic cleavage of the regulatory peptides. The substrates of CD10/NEP in the kidneys are (see the list of abbreviations) ANP, adrenomedullin and PAMP; in the brain, the substrates are enkephalins and oxytocin; in the lung, bombesin, BLP, GRP, neuromedin C, substance P and neurokinin A; in the cardiovascular system, angiotenisin II, bradykinin and CGRP; in the gut, VIP; on the neutrophil membrane, fMLP etc. Some substrates are not strictly tissue-specific, e.g. substance P. Preclinical and clinical trials explore possibilities of therapeutic application of the inhibitors of neutral endopeptidase, such as thiorphan in the management of pain, diarrhoea, depression, arterial hypertension and asthma. Other possibilities of application include the treatment of hyalinomembranous disease and prevention of neurotoxicosis in tetanus and botulism. PMID- 9748793 TI - Principles of the photodynamic therapy and its impact on the immune system. AB - Photodynamic therapy (PDT) is an encouraging procedure for treatment and diagnosis of a wide variety of malignant processes. There are three main mechanisms by which tumors are destroyed during PDT: direct cell destruction, damage to the vascular stroma and immune elimination. PDT of high intensity, targeted to a large number of cells results in immunosuppression. Low dose/energy or localized PDT stimulates the immune response against the tumor. The review reports recently performed animal experimental studies as well as the first human clinical studies. PMID- 9748794 TI - Prenatal determination of fetal RhD (rhesus positive) type by an amplification of DNA. AB - Examination of RhD genotype (Rhesus D gene) from amniotic cells (or chorionic villi cells) by PCR amplification of DNA can be done at early stage of pregnancy. Due to some missing exons in the Rh partial D variant (e.g. DVI), it is necessary to use different PCR systems to get relevant results. The localization of primers in our three PCR systems is on the different exons (10, 7, and 4 + 5). The advantage of PCR technique is prenatal detection of RhD of fetus from nonerythrocytes suspension (e.g. from an amnion fluid cell sediment) in comparison to a "standard" haemagglutination serological technique which uses blood erythrocytes only. The possibility of this technique to distinguished the heterozygous (D/d) or homozygous (D/D) fathers can help the clinicians in decisions about the management of further prevention of the hemolytic disease of newborn. PMID- 9748795 TI - Dent's disease--the hypercalciuric variant of Fanconi's syndrome. AB - Dent's disease is a rare type of proximal renal tubular defect characterized by hypercalciuria, low-molecular-weight (LMW) proteinuria, nephrocalcinosis and slowly progressive renal failure, short stature and osteopenia in children with clinical symptoms of rickets. This "hypercalciuric rickets" was originally described by Charles Dent and Max Friedman in 1964 [1]. The disease is probably linked to the X chromosome so that males are much more severely affected than females. PMID- 9748796 TI - [Comparison of adjuvant chemotherapy in breast carcinoma with a combination of cyclophosphamide, methotrexate, 5-fluorouracil (CMF) and AC (doxorubicin, cyclophosphamide). Initial results of a national cooperative study]. AB - The aim of this multicentric, prospective randomized trial is to evaluate and to compare, effects and toxicities of two chemotherapeutic combinations (AC and CMF) in adjuvant treatment of breast cancer. Both combinations were given in equitoxic doses and number of cycles was only four. There are 106 women treated for breast cancer T1c-3a, N0-1, M0 in the study. After surgery the patients were randomized, 54 for AC combination and 52 for CMF. We evaluate toxicity of this treatment in all patients in the study. Hematological and nonhematological side effects were comparable in both groups except alopecia (in the group AC was 100%). The study is not finished yet. Preliminary analysis does not show any difference between these two groups. PMID- 9748797 TI - [Interactions of cytokines with their receptors in the regulation of hematopoiesis, clinical and diagnostic applications]. AB - A short review on the interaction of hematopoietic growth factors with their receptors in the regulation of hematopoiesis. The introductory notes on the nature of cytokines and cytokine receptors, their biological and physicochemical properties as well as clinical and diagnostic consequences are mentioned. PMID- 9748799 TI - [Emergency medicine--who is the winner and who is the loser?]. PMID- 9748798 TI - [Risk factors for ischemic heart disease in patients with chronic fatigue syndrome]. AB - Risk factors of coronary artery disease (CAD) between a group of patients suffering of chronic fatigue syndrome (CFS) and a control group of healthy persons (whose exercise activity was not health-limited) were compared. Thirty three patients (27 women, 6 men, average age 39.9 +/- 11.7 years) and the same number of controls matched in age (39.8 +/- 10.3 years), gender and body weight. The Minnesota Questionnaire (by Taylor) and the Compendium of Physical Activities (by Ainsworth) were used to estimate total energetic expenditure in exercise activity as well as in job. The risk factors of CAD in the patients with CFS were not higher than in the control group. Aerobic physical fitness, basic anthropometric data, blood pressure, spectrum of blood lipoproteins, blood uric acid and smoking habits were not different between the compared groups. Patients suffering from CFS had lower total energetic expenditure in exercise activity. Nevertheless, this significant difference in sports activity was not large enough to cause any difference in risk factors of CAD between the CFS patients and the control group. PMID- 9748800 TI - [Pathogenesis of Helicobacter pylori infection]. PMID- 9748801 TI - [Gonorrhea--from a public health problem to a rarity]. PMID- 9748802 TI - [Should the oldest old be surgically treated?]. PMID- 9748803 TI - [Selective serotonin uptake inhibitors and interactions]. PMID- 9748804 TI - [Mortality resulting from femoral neck fractures in Norway 1980-94]. AB - Based on information obtained from the central register of death certificates held by the Norwegian National Bureau of Statistics trends in mortality resulting from fractura colli femoris were analysed for the period 1980-94. Mortality decreased by 30% (95% CI; 26%-34%) among women aged 60-98 years, whereas it remained constant among men. For both genders the risk of death from fractura colli femoris increased with age by approximately 20% per year. These changes are discussed in relation to the alleged epidemic of fractura colli femoris. PMID- 9748805 TI - [Ten years of the Lubinus Interplanta hip prostheses]. AB - This study presents the results of 332 Lubinus Interplanta total hip arthroplasties implanted in the period 1987-96 at the County Hospital of Laerdal, Norway. Kaplan-Meier survival curves based on reoperations reported to the Norwegian Arthroplasty Register were completed with the aid of a questionnaire to all patients, and by clinical and radiographic re-examination of the most symptomatic hips. The estimated survival of Lubinus Interplanta versus aseptic loosening was 99.6% after five years and 98.7% after ten years. The total revision rate was 0.7% after five years and 1.6% after ten years. Dawson's questionnaire demonstrated a very substantial improvement in quality of life and functional ability one to ten years postoperatively. Patients' perceptions demonstrated that indications for surgery and the results gained were similar to results obtained by other investigators. The least successful number (18%) of arthroplasties from a subjective point of view were reexamined, but the survival rate after re-examination showed no significant discrepancy compared with results based on figures from the Norwegian Arthroplasty Register. Despite the good results, changes in the choice of materials and routines are recommended. PMID- 9748806 TI - [Femoral shortening osteotomy for chronic hip dislocation in patients with cerebral palsy]. AB - Hip dislocation of several years duration in cerebral palsy needs treatment only if the patient has serious complaints. With the aim of reducing pain and problems with sitting function and perineal hygiene, we performed shortening osteotomy of the femur in 15 patients (12 girls and 3 boys) with spastic quadriplegia or diplegia at mean age of 14 (8-26) years. The patients were severely mentally and physically retarded, and only one patient had gait function, with support. A subtrochanteric shortening osteotomy of 3-5 cm was performed. The mean follow-up period was 5 (1-10) years. The symptomatic effect of the operation was good. The patients and parents were satisfied because the pain disappeared and the patients had less spasticity and stiffness. Complications were seen in two patients in the form of skin necrosis under both heels; this was caused by the plaster. Although reduction of the dislocation was not the aim of the surgery, radiographs at follow-up of 16 operated hips showed that five hips were reduced, whereas 11 hips remained subluxated or dislocated. We conclude that shortening osteotomy of the femur produces good symptomatic effects, probably due to reduction of the abnormally high muscle tension across the hip joint. PMID- 9748807 TI - [Decentralized high-dose cytostatic treatment with autologous stem cell support]. AB - In the past high-dose chemotherapy with autologous stem cell support in the treatment of certain types of cancer, was centralized to two hospitals in Norway. Almost three years ago it was decided that the treatment should be offered by all five university hospitals. In the northernmost university hospital of Norway, Tromso, peripheral stem cells were harvested from 29 patients after successful mobilization with chemotherapy and granulocyte colony-stimulating factor (G-CSF). After high-dose chemotherapy, more than 2 x 10(6) CD34-positive stem cells/kg were transplanted in 24 patients and a sign of reconstitution of bone marrow function was achieved with mean time for neutrophils > 0.5x10(9)/l, 9.8 days and for platelets > 20x10(9)/l, 10.8 days. No treatment-related deaths have occurred. Transplantation of selected CD34-positive stem cells has been performed in one patient. Recovery was comparable to the recovery of patients who had undergone transplantation with unselected products. This indicates that even small centres performing as few as ten procedures per year may offer high-dose chemotherapy with autologous stem cell support safely and successfully. PMID- 9748808 TI - [Gonorrhea caused by fluoroquinolone resistant gonococci]. AB - We describe a case of 4-fluoroquinolone resistant gonorrhoea where treatment failed on several occasions and where there may have been reinfections. Since 1995 peroral single dose 4-fluoroquinolones have been recommended as first preference treatment for uncomplicated gonorrhoea in Norway. However, Neisseria gonorrhoeae has a high ability to create resistance, and the emergence of 4 fluoroquinolone resistant strains is causing worldwide concern. An increasing number of 4-fluoroquinolone resistant gonococcal strains have also been observed in Norway, and the number of failures in the recommended treatment regime are increasing. It is proposed that 4-fluoroquinolone resistant strains be reported nationally. PMID- 9748809 TI - [Prognostic factors in squamous cell carcinomas of the head and neck]. AB - A review of prognostic factors in oral, pharyngeal and laryngeal carcinomas is presented. Distinctions are made between different kinds of prognostic predictors: external, host, and tumour predictors. The latter includes localization, stage, histological grading, DNA ploidy and biochemical characteristics. Finally, ideas on how best to utilize this knowledge and the implementation of any prophylactic measures are detailed. PMID- 9748810 TI - [Physiopathology of Helicobacter pylori infection]. AB - The gastric juice of Helicobacter pylori-infected individuals contains substantially higher levels of phospholipase A2 (PLA2) than that of individuals who are not infected. We present a new theory for how this H. pylori-induced PLA2 activity in gastric juice may play a major role in the development of peptic ulcer disease. When activated at neutral pH (pH 6.5-7.0), PLA2 may damage the surfactant-like, phospholipid-rich layer which constitutes an important part of the mucus barrier. Pepsin and other proteases, activated at low pH (pH 1.0-3.5), may then denature and cleave PLA2-exposed proteins. Peptic ulcers therefore tend to develop in regions exposed to changing luminal pH, such as the duodenal bulb when acid production is high or normal, or in the stomach when acid secretion is low. PMID- 9748811 TI - [New diagnostic criteria in Marfan syndrome]. AB - Marfan's syndrome is a relatively frequent autosomal dominant condition which is due to structural or quantitative changes in a connective tissue protein, fibrillin. The syndrome is associated with life-threatening changes in the aorta and serious manifestations in many different organ systems. Unclear diagnostic criteria and lack of use of the criteria in clinical practice may have led to overdiagnosing this syndrome in individuals with a long and slender habitus. This in turn can lead to negative consequences for both the individual and his or her family. Failure of diagnosis may cause even more harm, in particular because of the risk of sudden cardiac events. In 1996 an international group of experts proposed a set of revised criteria for Marfan's syndrome which takes into account molecular findings and family history (the Gent criteria). It is important that all practising physicians are aware of these criteria in order to prevent over- and underdiagnosing. A correct diagnosis is of major importance for medical follow-up, genetic counselling, habilitation, and counselling with regard to education and occupation. PMID- 9748812 TI - [High-resolution computer tomography of the lungs]. AB - High-resolution CT (HRCT) is a technique developed over the last decade. It optimises spatial resolution and provides details similar to those obtainable from gross pathologic specimens. HRCT of the lungs provides an accurate assessment of the pattern and distribution of many disease processes that in the conventional chest radiograph are occult or non-specific. HRCT is an established technique and the method of choice for evaluating a variety of pulmonary diseases. In this review we describe the modifications in CT technique that are instrumental in obtaining HRCT, and we also give examples of normal and pathologic findings in general. PMID- 9748813 TI - [Drug-induced hepatic injuries]. AB - This article provides a survey on various aspects of drug-induced hepatic dysfunction, including practical considerations for handling patients with suspected drug-induced liver affection. The assessment of causality is discussed on the basis of liver enzyme abnormalities, the onset and course of the reaction, and the response to readministration of the drug. Issues on when to withdraw a suspected drug and how to prescribe potentially hepatotoxic drugs to patients with liver diseases are also presented. PMID- 9748814 TI - [Interaction between sumatriptan and selective serotonin uptake inhibitors]. AB - Migraine is common among patients who suffer from depression, and this category of patients often needs drug treatment for both diseases. A pharmacist consulted the Regional Drug Information Centre in the western part of Norway (RELIS 3) about the combined use of sumatriptan and fluoxetine, as the product information on sumatriptan warns of a possible interaction between these drugs. This possibility was evaluated by the Drug Information Centre, and it was concluded that the combination of sumatriptan and a selective serotonin reuptake inhibitor is not contraindicated. This conclusion was based on both theoretical and clinical considerations. PMID- 9748815 TI - [Integrated biology--the future biomedical science]. PMID- 9748816 TI - [House calls should not be a rarity]. PMID- 9748817 TI - [House calls strictly on significant medical indications]. PMID- 9748818 TI - [Peroral anticoagulant therapy]. PMID- 9748819 TI - [Cholesterol levels in Norwegian adolescents]. PMID- 9748820 TI - [Development of smoking habits in Norway]. PMID- 9748821 TI - [Follow-up after colorectal cancer. Undeserved neglect of CEA-analysis]. PMID- 9748822 TI - [Serum cobalamin levels in elderly psychiatric patients with depression and dementia]. PMID- 9748823 TI - [Adverse effects of zopiclone]. PMID- 9748824 TI - [Physicians as executioners]. PMID- 9748825 TI - [Quality requirements also for publications on alternative medicine]. PMID- 9748826 TI - [The Tidsskrift also for students?]. PMID- 9748827 TI - [The internship is a part of basic medical education--improvement urgently required]. PMID- 9748828 TI - [Rehabilitation as regional policy practice]. PMID- 9748829 TI - [Use of risk number in chronic diseases. Number needed to cheat?]. PMID- 9748830 TI - [Significant challenges in neurology--also for health administrators!]. PMID- 9748831 TI - [Attitude to and practice of Norwegian gynecologists concerning hormone replacement therapy in climacteric]. AB - A questionnaire was sent to all 475 members of the Norwegian Gynaecological Society. It was based on a similar study previously performed in Denmark and Sweden. 85% of the members returned the questionnaire. 382 (80%) had answered the questions; 153 (40%) women, 228 (60%) men, and one case where the sex was not stated. The mean age was 48 years (SD 10). The male gynaecologists had a more liberal attitude towards hormone replacement therapy than their female counterparts, 43% of them recommending oestrogen for all women, compared to 31% of the female gynaecologists. The younger doctors were more restrictive in their recommendations but attitudes became more liberal the older the doctors were. Among gynaecologists over 55 years, 49% of males and 50% of females recommended oestrogen for all women. The final decision as to whether or not to take hormone replacement therapy was most often made by the patient herself (61%). The majority of both female (86%) and male (75%) gynaecologists considered risk factors for heart disease to be an indication for oestrogen. In perimenopausal women, 356 (93%) preferred oral cyclical oestrogen combined with progestagen, whereas in postmenopausal women 333 (87%) preferred to take oral oestrogen combined with progestagen continuously. PMID- 9748832 TI - [Use of hormone replacement therapy among female gynecologists and partners of male gynecologists in Norway]. AB - In 1997 a questionnaire on hormone replacement therapy was sent to all 475 members of the Norwegian Society for Obstetrics and Gynaecology. There was an 80% response rate. Among the questions asked, were some concerning the members' personal use of hormones. 36 of the 153 female gynaecologists were menopausal or had climacteric complaints. 28 (78%) out of the 36 used oestrogen, four (11%) had contraindications, and another four (11%) were not in need of therapy. Of 228 male gynaecologists 96 had partners who had either climacteric symptoms or were menopausal, and 74 (77%) of these were receiving hormone treatment. The reasons given for not using oestrogen were lack of symptoms requiring therapy in 14 cases (15%) and contraindications in one case (1%). In seven cases (7%) no information was provided. In 1996 about 19% of Norwegian women aged 45 to 69 years used hormone replacement therapy. The use of hormones among female gynaecologists and partners of male gynaecologists is thus four times higher than in the rest of the population. PMID- 9748833 TI - [Physicians and other practitioners of acupuncture in Norway--education, theoretical orientation and practice]. AB - We conducted a survey for the purpose of obtaining information on training, theoretical orientation and practice among various categories of practitioners of acupuncture in Norway. Particular attention was paid to physicians, compared with other groups of practitioners. A questionnaire was sent to 161 persons who had attended Norwegian Medical Acupuncture Seminars (Norske legers akupunkturkurs) and to 274 persons found under "Acupuncture" in the Yellow Pages of the telephone directory. The response rate was 80%, questionnaires being returned by 298 practitioners. A significantly higher percentage of physicians, as opposed to other practitioners, had less than 120 hours of acupuncture training. 30% of the physicians, compared to 60% in other groups, had more than 10 acupuncture consultations a week. Physicians performed only a relatively small number of different acupuncture methods. There was a relatively high number of men among the acupuncturists. Hence, acupuncture seems to be a practice with masculine appeal. 67% of the physicians used one or more Chinese medical concepts associated with acupuncture. 45% stated that they found non-scientific explanations for how acupuncture works reliable. We therefore argue that one can identify parallel processes in the development of acupuncture. Some elements of acupuncture have been integrated in a scientifically defined reality. At the same time, this survey indicates another process: it shows that many practitioners use traditional Chinese medical concepts. This may indicate that some practitioners have changed their view on what constitutes a reliable picture of reality. PMID- 9748834 TI - [The annual neurological conferences 1990-97--extensive scientific presentation for Norwegian neurologists]. AB - 324 free oral presentations were put forward during the Norwegian annual neurology conferences in 1990-1997. The lead authors of 216 presentations (67%) were employed at university hospitals; 108 authors were in employment at other hospitals or institutions. 159 lead authors (49%) were specializing in neurology and these candidates became increasingly active as presenters during the period in question. International collaboration occurred in 27 of the presentations (8%), nine of the presenters working with colleagues in the USA and seven with colleagues in Sweden. The presentations were classified as either mainly clinical, with 275 presentations (85%), or mainly basal science, with 31 presentations. In the remaining 18 presentations administrative or historic literary topics were discussed. The disorders most frequently dealt with were epilepsy (88 presentations), cerebrovascular disorders (52 presentations), and autoimmune disorders (47 presentations). There was an increasing trend towards the latter two topics during the conference period. In conclusion, this survey shows neurology to be an expansive specialty with a focus on active development and research. PMID- 9748835 TI - [Narcolepsy in children--a diagnostic and therapeutic challenge]. AB - Narcolepsy is a socially and psychologically disabling disease that most often develops in adolescence or early adulthood. In a number of studies about one third of the patients had experienced the first symptoms before the age of 15. The diagnosis and subsequent treatment is usually not established until several years later, often ten to fifteen years after the appearance of the first symptoms. If unrecognized and untreated, narcolepsy may lead to serious psychological and social problems during childhood and early adulthood, which may in turn cause difficulties in social adjustment later in life. This stresses the importance of early diagnosis. The diagnosis is based on clinical and polysomnographic criteria. In children, however, the clinical symptoms and polysomnographic findings may be atypical, making it difficult to establish a definite diagnosis. Three patients, two five-year olds and one four and a half year old, are presented to illustrate the problems and considerations which must be taken into account in the diagnosis and treatment of children with narcolepsy. PMID- 9748836 TI - [Should treatment of risk factors of cardiovascular diseases be directed only by absolute risk?]. AB - Consensus groups have recommended using the baseline absolute risk of disease over five to ten years rather than relative risk when treatment with lipid lowering or antihypertensive drugs is considered. Targeting patients with a coronary event rate of 20% over ten years ensures that society's investment in drugs yields substantial benefits by reducing the incidence of premature disease. However, the use of absolute risk is most appropriate among the middle-aged. Because all elderly persons have a high absolute risk of disease, almost all may be eligible for drug treatment. The costs of postponing death after the age of 70 are therefore substantial. Among young persons with familial hypercholesterolemia on the other hand, the consequences of a death before middle-age are so enormous that most patients are treated with drugs even though absolute risk is very low. An additional problem with using absolute risk is that the risk of atherosclerotic disease accumulates over time. Calculation of benefit on the basis of short-term trials might underestimate long-term benefit. In the future, non-invasive measures of atherosclerosis and new markers of risk may provide valuable information on risk stratification in primary prevention. PMID- 9748838 TI - [Practice is learned by practice]. AB - Efficient continuing education for experienced physicians should build upon and reflect doctors' concrete everyday experiences and self-defined learning needs. Reciprocal practice visits among general practitioners, afford excellent opportunities for colleagues to see and be seen, to reflect, discuss, learn from each other and develop networks, using commonly experienced consultations as a starting point. Practice visits have been credited within the Norwegian continuing medical education system since 1989, but very few practitioners have made use of the method. We have developed a course where groups of 10-12 experienced general practitioners conduct four whole day practice visits over a 2 month period. Group sessions, totalling 20 hours, are used to develop cohesion and trust among colleagues through discussions and exercises. In the later stages, plenary sessions are used to present and discuss experiences from all of the practice visits, and participants are encouraged to develop ideas and strategies concerning their professional future. The participants in the three courses arranged so far (n = 31) were general practitioners with, on average, ten years of experience. Evaluations were positive, and emphasized the value of the course as an opportunity to break the relative isolation of general practice, and share experiences, insights, frustrations and ideas with colleagues. PMID- 9748837 TI - [Female sex hormones increase the risk of breast cancer]. AB - The incidence of breast cancer in women in increasing, partly due to changes in age distribution in the population, and partly due to a real increase in risk. Changes in family patterns may, to some extent, explain the increased risk since giving birth to a first child late in life and bearing few children both increase the risk of breast cancer. The influence of female sex steroids on the breast plays a central role, but the biological mechanism is not clearly understood. There is a certain amount of risk involved in using hormonal medication (oral contraceptives or postmenopausal hormone replacement), but on ceasing to take the medication, risk will revert to the expected rate within a few years. Future epidemiological research on breast cancer will concentrate on events occurring during hormonally potent phases of life, such as growth and development during the fetal period, and sexual and somatic maturation during adolescence. Until now only modest interest has been shown in researching these two particular phases, but both may be important for the natural course of breast cancer. PMID- 9748839 TI - [How does professional competence develop?]. AB - The reality of medical practice demands not only proficiency but also educational provisions, an aspect which traditionally has been given little attention in medicine. The competence of a general practitioner does not consist solely of professional knowledge and practical skills; it consists to a large extent of personal (intuition and creativity based on personal experience) and communicative elements, both of which are necessary in managing the unpredictable complexity and the many controversies of general practice. Such competence is difficult to develop outside the practice. By sharing the experience of actual consultations with experienced colleagues, and reflecting upon this experience within a small group, general practitioners will probably develop better professional judgement and increased awareness of their learning needs, as well as of their limitations and potential. This article discusses these topics using the authors' personal experiences in the continuous medical education effort which is based on reciprocal practice visits among a small group of experienced general practitioners. PMID- 9748840 TI - [Electronic house call]. AB - Telemedicine technology will soon be widely available in physicians' surgeries and patients' homes. In this article we discuss the broader implications of its widespread use among physicians, patients, and society as a whole, focusing on the main vehicle of communication between physician and patient; the "electronic home visit" or "electronic house call". After exploring the new relationship that is likely to develop between patients and physicians as a consequence of the use of telemedicine, we then discuss the anticipated broader social, economic and legal consequences of the widespread use of this technology. PMID- 9748841 TI - [Hemochromatosis--molecular genetic screening of blood donors?]. PMID- 9748842 TI - [Why aren't the Norwegian clinicians doing their research abroad?]. PMID- 9748843 TI - [Advantages of problem-based learning]. PMID- 9748844 TI - [Problem-based learning. Free statements of a small and insignificant but strong opposition]. PMID- 9748845 TI - ["Too HOT" blood pressure?]. PMID- 9748846 TI - [Iron balance and boys during puberty]. PMID- 9748847 TI - [Who is to define non-beneficial treatment?]. PMID- 9748848 TI - [Sick-listing based on failing basis?]. PMID- 9748849 TI - [Lumbar disk prolapse and chemonucleolysis]. PMID- 9748850 TI - [Down or synthetic stuffings in pillows--what is better?]. PMID- 9748851 TI - [Prehospital thrombolysis in acute myocardial infarction]. PMID- 9748852 TI - [Development of district psychiatric service in Denmark]. PMID- 9748853 TI - [The Internet and psychiatry]. PMID- 9748854 TI - [International health. A vision on more equal possibilities for more people to achieve a healthy and meaningful life]. PMID- 9748855 TI - [Familial hemiplegic migraine]. AB - Familial hemiplegic migraine (FHM) is a rare subtype of migraine with aura. It is inherited as an autosomal dominant trait. A gene for FHM has been assigned to chromosome 19. This gene codes for a brain-specific calcium channel, and is responsible for FHM in 55% of the FHM families. Other FHM families have been linked to two different locations on chromosome 1. These locations contain possible candidate genes coding for calcium-and potassium channels. Thus FHM is a genetically heterogenous ion channel disorder, which is caused by at least three different genes. About 29% of the FHM families also have cerebellar ataxia, these families have all been linked to chromosome 19. The identification of the genes for FHM may be a key to the identification of the gene/genes for migraine with and without aura. PMID- 9748856 TI - [Fragile X syndrome. Diagnosis, genetics and clinical findings]. AB - The most common heritable form of mental retardation is the fragile X syndrome. It is X-linked and affects 1:1500-1:4000 boys. In Denmark 230 affected individuals are known, thereby rendering it underdiagnosed. Only minor dysmorphic traits are associated with the syndrome, more pronounced in boys, namely a long face with large, prominent ears, and macroorchidism postpubertally. The psychological manifestations are autistic features, hyperactivity and deviant behavior. Today no medical curative treatment is available but much can be gained from social and educational intervention. The syndrome is caused by a dynamic mutation on the X chromosome and displays a remarkable pattern of inheritance. Carrier diagnosis and prenatal diagnosis are feasible. This article describes the clinical, diagnostic and genetic aspects of fragile X syndrome. PMID- 9748857 TI - [Effects of establishment of community psychiatry in Aalborg]. AB - The aim of the present study was to illustrate the effects of community-based psychiatry. The catchment area was divided into three homogeneous districts, East, North and West. Teams were established on 1.9. 1990, 1.10.91 and 1.5.1992, respectively. Social, diagnostic and treatment related data were gathered from two cross-sectional investigations (I: February 1992 and II: February 1993) and from in-patients and out-patients files. In cross-section I a majority of long term ward patients and hospital-based employment offers was found in the district where the community district team had not yet been established. In the district where the first community district team was established most primary target patients were treated. In cross-section II the hospital-based psychiatric service were more homogeneously distributed between the districts. The establishment of community-based psychiatric teams resulted in new referrals, and increasing numbers of patients becoming attached to the psychiatric teams, but crowding and use of compulsory measures in hospital also increased. PMID- 9748858 TI - [Variations in hospital based psychiatric service between Aalborg and Aarhus. A registry-based 10-year investigation]. AB - The study was based on data from the Danish Psychiatric Central Register. It consisted of 11,753 persons with permanent residence in the municipalities of Aalborg or Arhus, who had 32,557 admissions to a psychiatric hospital in Denmark during the period 1982-1991. The admission rates declined by 30%. The involuntary admission rate increased during the later years of the period, it was higher at the Psychiatric Hospital in Arhus (PHA). The bedday use per admission was 5.5 weeks on average at Aalborg Psychiatric Hospital (AAPH) and nine weeks on average at PHA. Through the period the bed rate (beds in use) was gradually reduced by 40%, most significantly at PHA. It can be concluded that the treatment at AAPH was less time-consuming. This is mainly caused by there being fewer beds available. Indices for quality of outcome did not give reason for conclusions in regard to differences in quality of treatment. PMID- 9748859 TI - [Percutaneous transluminal coronary angioplasty in acute myocardial infarction. A prospective controlled study]. AB - In the present study we compared the outcome of primary percutaneous coronary angioplasty (PTCA) (PTCA without prior or concomitant administration of thrombolytic drugs) in 82 consecutive patients with acute myocardial infarction (AMI) with the outcome of 82 AMI patients, who were treated with intravenous thrombolysis. The thrombolysis patients were prospectively matched to the angioplasty patients regarding age, sex, duration of symptoms and infarct localisation. The in-hospital mortality was 3.7% in the PTCA group versus 4.9% in the thrombolysis group. Thrombolysis-treated patients had increased use of diuretics and ACE-inhibitors as compared to PTCA-treated patients. The mean ejection fraction was 52 +/- 11% in the PTCA group versus 47 +/- 10% (p = 0.01) in the thrombolysis group. We conclude that initial Danish experience with primary PTCA is promising, and that this treatment may favourably affect the outcome of acute myocardial infarction. PMID- 9748860 TI - [Can radioactive iodine be used in the treatment of diffuse non-toxic goiter?]. AB - Traditional treatment modalities of diffuse nontoxic goitre are thyroid hormone suppression or surgery. When treating nodular nontoxic goitre with 131I treatment, a reduction in thyroid volume to about 50% is seen. In the present study we evaluated the effect of 131I treatment in 21 patients treated for a diffuse nontoxic goitre and followed by evaluation of thyroid volume measured by ultrasound. Thyroid volume declined in all patients from median of 66 ml (range 27-160 ml) to 21 ml (9-108 ml) over a year, a reduction of 62%. Three patients developed hypothyroidism in the follow-up period (14%), one of these had a temporary hyperthyroid fase. In conclusion, 131I treatment of diffuse nontoxic goitre reduces thyroid volume by approximately 60% within 12 months. Hypothyroidism developed in 14% during a limited follow-up period. PMID- 9748861 TI - [Bipolar affective disorder. A retrospective study of 158 patients in a well defined geographical region]. AB - A cohort of 158 patients was identified from 723 patients admitted in 1990 or treated as outpatients in 1991 for affective disorders in three university hospitals in Copenhagen, covering a well-defined catchment area. The cohort was subclassified for seasonal pattern according to DSM-III-R. Meteorological data of temperature, hours of sunshine, rainfall and wind-velocity expressed as mean values/month were obtained from the Institute of Meteorology in Copenhagen. There was no relationship between the annual dis- tribution of affective episodes and meteorological data, neither in the total cohort nor in the subgroup of patients with seasonal patterns The present study does not support the hypothesis that a relation exists between start of a new episode and the season or climatic condition. PMID- 9748862 TI - [Incidence of child neglect and child abuse in the region of Copenhagen]. AB - The aim of the study was to evaluate the incidence of physical violence, neglect and sexual abuse against children as reported to the local authorities, in the county of Copenhagen during the year 1993. A questionnaire was mailed to the local authorities in the 18 districts in the county and to 18 schools and 18 general practitioners (GP's). They were asked whether, and how many new, verified and suspected cases of physical, emotional and sexual abuse had come to their attention in 1993. A total of 300 cases of child abuse were identified in the area with a total population of 675.000 i.e. a total incidence of 2.7 cases per 1000 children aged 0-17 years (2.7%), with 0.7% for physical violence, 0.5% for sexual abuse and 1.5% for emotional neglect. Some 180 cases were considered verified and 120 cases were suspected. A considerable variation between the different districts was noted, i.e. the local incidence varied from 0-5.4%. A statistically significant inverse correlation between the average local tax income and the incidence of child abuse and neglect was found. There were several examples of schools having knowledge of more cases than the local authorities. Very few cases came to be attention of the GP. PMID- 9748863 TI - [SeHCAT scanning in bile acid malabsorption]. AB - Chronic diarrhea caused by bile acid malabsorption (BAM) is usually divided into three groups. Type 1 is associated with ileal disease or ileal resection; type 2 is idiopathic, and type 3 is BAM associated with certain predisposing conditions. We evaluated the applicability of the SeHCAT test as a routine investigation of different types of suspected BAM. Detailed information about 298 patients were obtained from retrospective review of patient records. All 68 patients with ileal resections had abnormal SeHCAT retention (median 0.6%; range 0-13%). Of 42 patients with non-resected Crohn's disease or radiation injury, BAM was found in 28 cases. A diagnosis of BAM type 2 was established in 33 of 150 patients with unexplained chronic diarrhoea. For patients tested for possible BAM type 3, the SeHCAT values were significantly lower compared to type 2 patients. For BAM type 1, the SeHCAT test is only recommended in non-resected patients. Idiopathic BAM seems to be more common than recognized. The presence of certain predisposing conditions might strengthen the indication for SeHCAT testing. PMID- 9748864 TI - [Benign paroxysmal torticollis. Recurrent involuntary twisting of the head in infants and young children]. AB - Benign paroxysmal torticollis occurs in infancy and early childhood. The etiology is unknown, although a vasomotor labyrinthine pathophysiology is possible. We report four cases with onset at ages of two to six months of recurrent episodes of torticollis and discomfort persisting between three hours and seven days. In three cases the torticollis was alternating. Photography or videorecording made by the parents during the attacks were helpful for the diagnosis in three cases. MRI may in some cases be necessary to rule out a space occupying lesion in the posterior fossa. PMID- 9748865 TI - [Hepatitis C virus transmission between two brothers]. AB - We present a case where a 30 year-old male by accident probably infected his older brother with hepatitis C-virus. After two months this older brother suffered from acute hepatitis C, and within 16 months he showed progression to chronic hepatitis C. PMID- 9748866 TI - [Aging and the elderly. New answers to old questions]. PMID- 9748867 TI - ["Prime-MD"--a quick method for diagnosis of mental disease in general practice. Is the documentation correct?]. PMID- 9748868 TI - [The curse of ticks]. PMID- 9748869 TI - [Comment on a critical survey of a Cochrane review on the treatment of tonsillitis]. PMID- 9748870 TI - [A status article on the effect of antibiotics in tonsillitis]. PMID- 9748871 TI - [Contribution to statins?]. PMID- 9748872 TI - [Changes in thrombocyte aggregation in primary hypothyroidism]. AB - The authors examined the thrombocyte aggregation in 10 controls and 17 patients with the diagnosis of primary hypothyroidism before and after 2 months substitution treatment with levothyroxine. They recorded a significantly reduced intensity of the aggregation response in untreated patients as compared with controls after adrenaline (p < 0.01), ADP (p < 0.01) but not after ristocetin. Impaired thrombocyte aggregation was observed in 11 of 17 patients, i.e. in 65%. After treatment the thrombocytopathy improved in 7 of 11 patients (63%), in four it persisted. Except one female patient the thrombocytopathy improved in all patients with manifest hypothyroidism. In patients with the latent form of hypothyroidism probably an independent coincidence of elevated TSH levels and impaired thrombocyte function was involved. The authors did not detect any cases of acquired von Willebrand's disease. In the conclusion the authors mention that impaired thrombocyte aggregation is a frequent phenomenon after thyroxine treatment. It may be of clinical significance when combined with other changes of haemostasis or in conjunction with the use of some drugs. PMID- 9748873 TI - [Treatment of endocrine orbital disease by elimination of the thyroid gland. The effect of unsuccessful elimination, severe pretibial myxedema and persistence of hypothyroidism after removal in long-term results]. AB - The reason why soon after elimination of the thyroid gland protrusion of the bulbi does not recede and adequate regression of soft orbital tissue infiltration does not occur is at first the short time which has elapsed after elimination. The result usually is recorded only later. The long-term cause of minor improvement is the impossibility to combine surgery of the thyroid gland and radioiodine with corticoids, as well as incomplete thyroid elimination and presence of residues and finally pretibial myxoedema associated with a persisting high level of antibodies against the receptor for TSH. Hypothyroidism after the eliminating dose must not be left too long, so far a period of 3-4 weeks seems adequate. PMID- 9748874 TI - [Determination of cardiac troponin-I in prediction of thrombolysis]. AB - Acute myocardial infarction (AMI) is a disease with high morbidity and mortality. Diagnosis of AMI using common methods (classical biochemistry, ECG) fails even in the fifth part of patients so that other noninvasive diagnostic methods are preferred. Recently, the biochemical analysis has been restored in the case of AMI diagnostics and also in prediction of coronary reperfusion after administration of a fibrinolytic agent. A suitable markers of AMI diagnostics is a combination of myoglobin and cardial troponin-I which is reported as a marker with high specificity and sensitivity. To determine coronary reperfusion, the examination of cardial troponin-T and CK-MB mass is recommended. In the literature, there exist isolated papers dealing with dynamics of cTn-I suitable for prediction of coronary revascularization. However, these papers do not report any adequate algorithm and subsequently mathematical differences between successful thrombolysis and failing thrombolysis. Therefore the aim of our study was to describe dynamics of cTn-I changes in AMI patients treated by thrombolysis. The study comprised of 8 AMI patients with delay from the occurrence of pains to fibrinolysis application under 4 hours (delay 4 hrs). These probands were examined for concentration of cTn-I and CK-MB mass in 3-hour intervals in the first 48 hours after admission to the clinic and further in 6 hour intervals from the hour 48 to the hour 90 after admission. All probands had a successful reperfusion (estimated using CK-MB peak, in 4 patients reperfusion was verified by subsequent coronarography). However, a simple mathematical prediction of coronary reperfusion after acute myocardial infarction by means of cTn-I dynamics determination is not possible due to relatively low cTn-I differences in individual analyses (CK-MB mass analysis shows more significant differences). Thus, in order to determine coronary revascularization, we recommend to use common analyses of dynamics of cTn-T or CK-MB mass. PMID- 9748875 TI - [Levels of vitamins A, E and C in serum and gastric juice in relation to gastric mucosa and occurrence of Helicobacter pylori]. AB - Colonization of the gastric mucosa with Helicobacter pylori H.p. reduces the vitamin C concentration of gastric juice. Eradication of H.p. within four weeks after completed treatment does not exert a significant effect on changes in the concentration of vitamins A, E and C in gastric juice or serum. Despite this after eradication a rising trend of vitamin E in gastric juice was recorded. Substitution of vitamin C and E in gastritis associated with colonization with H.p. has a favourable effect and may reduce the risk of malignization. PMID- 9748877 TI - [The short-acting insulin analog Lispro (Humalog) in the treatment of diabetes- comparison of preprandial and postprandial administration and comparison with treatment using Humulin R]. AB - The results of a clinical trial comprising 162 type 1 and 2 diabetics who took for 12 weeks Humalog in cartridges revealed that administration of this insulin before or 20 minutes after a meal does not affect the blood sugar level in a major way. None of the patients developed after Humalog administration local or general allergic manifestations. Hypoglycaemic episodes grade I and II were not more frequent during treatment with human insulins. The mean compensation of diabetes (HbA1c) remained unchanged. 80% of the diabetics are statistically significantly satisfied with Humalog treatment as compared with Humulinem R administration. PMID- 9748876 TI - [Treatment of multiple myeloma with high-dose chemotherapy and transplantation of autologous hematopoietic stem cells and subsequent maintenance therapy with interferon alfa-2b or interferon alfa 2b and dexamethasone. Report of the ongoing study of the "4W" Czech Myeloma Group]. AB - We report our results with high-dose chemotherapy in previously untreated multiple myeloma patients (4 courses of VAD chemotherapy, collection of PBSC after priming with cyclophosphamide, 5 g/m2, high-dose chemotherapy with melphalan, 200 mg/m2). Second transplantation was indicated only for patients who did not achieve remission after the first high-dose therapy (paraprotein lower than 25% of the pretreatment value). For the second transplantation melphalan (200 mg/m2) with methylprednisolone (1.5 g for 5 days) were used as conditioning regimen. After high-dose therapy all patients were randomized into two arms of maintenance therapy: interferon alpha-2b or sequential maintenance therapy (interferon alpha-2b for 3 months followed after 4 week pause by 40 mg of dexamethasone days 1-4, 10-13 and 20-23. The administration of interferon alpha was resumed four weeks after the last dexamethasone for next three months. The maintenance therapy continued for 48 months or until the progression. Fifty-five patients were enrolled in the study from January 1996 to August 1997. Thirty-five patients have undergone the first transplantation and 57% of them reached complete remission. There were 10% of non-responders after the first high-dose regimen. The mean time to reach white blood cell count above 1 x 10(9)/L after the application of high dose melphalan and platelets more than 50 x 10(9)/L were 12.2 (range 6-16 days) and 12.4 (range 0-25 days), respectively. Grade 4 mucositis according to SWOG classification requiring total parenteral nutrition was presented in 40% of the patients. The mean number of 1 unit of platelets and 2 units of packed red blood cells transfusions were given within the posttransplant period. Early transplant related mortality was 3%. This paper describes the response and tolerance of each particular step of therapy. The follow-up has been too short to evaluate event-free and overall survivals. PMID- 9748878 TI - [What dosage is sufficient in combined inhalation therapy (fenoterol + ipratropium bromide) in patients with exacerbation of chronic obstructive lung disease?]. AB - The authors compared in a prospective study the bronchodilatating and undesirable effects of combined inhalation treatment (phenoterol + ipratropium bromide) in the treatment of patients with exacerbation of chronic obstructive pulmonary disease, using different dosages. The patients were divided at random into two groups--group one inhaled berodual sol 3.5 ml/day (i.e. 1.75 mg phenoteroli + 0.875 mg ipratropii bromidium), the second group had a dose of double size. During the trial the authors monitored the peak expiration rate, the heart and respiration rate, blood gases and the subjective state of dyspnoea, using a 10 cm line. By comparison of bronchodilatating and undesirable effects they reached the conclusion that a daily dose of 3.5 ml berodual sol. is sufficiently effective. Increasing the daily dose to 7 ml did not produce a greater therapeutic effect nor increase the risk of undesirable effects. PMID- 9748879 TI - [Needle-like inclusions in liver cells are specific diagnostic signs of porphyria cutanea tarda]. AB - Needle-shaped cytoplasmic liver cell inclusions are considered to represent a feature typical of porphyria cutanea tarda (PCT). Histological sections of 849 liver biopsies were stained with ferricyanide reduction reaction for demonstration of the inclusions. They were detected in 18 cases, and in all of them the diagnosis PCT was clinically and/or biochemically confirmed. In our group of patients, PCT was associated with alcoholic liver disease in three cases, with chronic hepatitis C in three cases and with hepatocellular carcinoma in two cases. In the liver with hepatocellular carcinoma, the inclusions were present only in non-neoplastic liver tissue, not in the tumourous tissue. No inclusions were found in the liver tissue of patients without clinical signs of PCT. PMID- 9748880 TI - [Schnitzler's syndrome]. AB - The authors describe the development of disease in a patient with monoclonal immunoglobulin IgM, urticarial morphoeae and intensive pain in the region of the pelvic bones due to osteolysis and osteosclerosis. The combination of these symptoms corresponds with the so-called Schnitzler syndrome which is analyzed in detail in the discussion. The urticarial manifestations diminish only temporarily during corticoid treatment, other drugs have no effect. 2-chlorodeoxyadenosine, the most effective drug in treatment of m. Waldenstrom, suppressed the skin manifestations temporarily. However, the concentration of monoclonal IgM did not decline even after two cycles and therefore the authors did not proceed with it during subsequent cycles. The severity of urticaria called for a permanent daily dose of Prednisone. The second symptom, bone pain, was however influenced by the administration of pamidronate (Aredia), using an initial dose of 120 mg per month. Now the patient takes a maintenance dose of 60 mg/month. The authors describe the case to draw attention to a rare cause of urticaria and the possibility to treat bone pain with bisphosphonates. PMID- 9748881 TI - [Controlling the progression of diabetic nephropathy]. AB - Diabetic nephropathy is a serious microangiopathic complication of type 1 and 2 diabetes and accounts in a major way for the high morbidity and invalidity of diabetics. Although progression is inevitable when the disease is advanced, there are ways how to prevent its development and retard its progress. Because reliable prediction of diabetic nephropathy is impossible, primary prevention is essential in all diabetic patients. As soon as the diagnosis of diabetes is established, the blood sugar level should be checked systematically and permanent satisfactory compensation of diabetes should be ensured (HbA1c less than 6.5%) which is the main principle throughout the subsequent course of the disease. The principle of secondary prevention the objective of which is to prevent the development of manifest nephropathy with permanent proteinuria, is to monitor microalbuminuria and maintenance of a normal blood pressure. With regard to pathophysiological circumstances in therapy angiotensin converting enzyme inhibitors are preferred. The objective of tertiary prevention is retardation of renal insufficiency by fortified hypotensive therapy, correction of hyperlipoproteinaemia, dietary protein restriction and adequate compensation of diabetes. In case of renal failure dialyzation treatment or transplantation must not be delayed. PMID- 9748882 TI - [Treatment of urinary infections in chronic renal insufficiency]. AB - The author describes some differences in the course of treatment of urinary pathway infections in prolonged reduced renal function. He emphasizes changes from the aspect of the clinical course of urinary infections and changes in the pharmacokinetics and pharmacodynamics of antibiotics and principles for selection of therapy are outlined. PMID- 9748883 TI - [ATP-sensitive potassium channels and substances affecting them]. AB - ATP-sensitive potassium channels are the site of action of the sulphonylurea derivatives that are used to treat non-insulin-dependent diabetes mellitus. These ATP-sensitive potassium channels are also found in myocardial cells and in vascular smooth muscle cells. The sulphonylurea derivatives have been reported to cancel the cardioprotective effects by blocking the opening of these channels in myocardium and vascular smooth muscles. A new sulphonylurea derivative, glimepiride, has been shown to be devoid of vascular ATP-sensitive potassium channel binding properties. The so called potassium-channel-openers, on the other hand, are expected to be used in the treatment of hypertension, ischemic heart disease and asthma bronchiale. PMID- 9748884 TI - [Clinical use of glycoprotein IIb/IIIa receptor blockers in invasive cardiology]. AB - Acute coronary syndromes (unstable angina pectoris, acute myocardial infarction) are still the main cause of mortality and morbidity not only in economically advanced countries. Although coronary angioplasty has become modern treatment of stable angina as well as of acute coronaries, acute occlusion of the vessel is still a serious problem. The prognosis of patients undergoing coronary angioplasty can be substantially improved by accurate evaluation of the risk of possible complications during or shortly after the procedure and by development of more effective anticoagulants. Research provided evidence that platelets play a decisive role in the formation of occlusive thrombi and that the thrombocyte glycoproteins IIb/IIIa are the basic mediator of platelet aggregation. Newly developed inhibitors of these receptors reduce acute ischaemic complications after percutaneous transluminal coronary angioplasty. Conclusions of various trials support the inclusion of glycoprotein IIb/IIIa inhibitors in the standard therapeutic protocol of patients undergoing coronary angioplasty, in particular those with a high risk because of acute occlusion of the vessel after operation. PMID- 9748885 TI - [The value of studies of quality of life in chronic respiratory tract diseases in the framework of holistic medicine]. AB - Quality of Life (QOL) is a new area of research evaluating the psychical, functional and psychological components of human life. According to current understanding of health issues the measurement of morbidity or mortality does not estimate the health status and describe the influence of illness on human life. The theoretical framework of Health Related Quality of Life is largely based on a multidimensional perspective of human as physical, psychological and social functioning and well-being, along the WHO definition of health. QOL assessment could be carried out using different psychological methods. There are several questionnaires developed to assess the quality of life in patients. The measurement instruments are generic (used in wide range of health related issues), domain specific (concerning some important outcome such as social support, coping), and disease specific which are used to assess patients with particular health problems. QOL assessment could be used as the measurements in pharmacoeconomics and clinical trials. Polish QOL Initiative Group produce Polish version of existing questionnaires--for example The Asthma Quality of Life Questionnaire was registered in Polish language version. Quality of Life researches facilitate doctors to understand patient's perceiving of the health state and help them to live a fulfilling life. PMID- 9748886 TI - Porownanie skutecznosci piracetamu i dekstranu 40 tys. u chorych w podeszlym wieku z niedokrwiennym udarem mozgu. AB - Piracetam is believed to restore the metabolism of glucose and 02, and also to prevent the vasospasm of small arteries in the penumbra of ischaemic focus. Taking the above into account we tried to estimate its effect in elderly patients with stroke. The studied group consisted of 47 patients, aged 60-78 years (mean 67,4 yrs), with the first ischaemic episode. 23 patients were treated with 500 ml of Dextran 40 daily and 60 ml of placebo given separately. The rest of the patients was treated with the same dose of Dextran 40 and 12 g of Piracetam as 60 ml i.v. bolus. The neurological status of the patients was estimated using Scandinavian Neurological Stroke Scale (SNSS). In the patients treated with Dextran and placebo there were no changes in the total score of SNSS, both after 10 and 28 days, comparing with the initial status. In the patients treated with Dextran and Piracetam total SNSS score improved significantly after 10 days (p < 0.05) and the effect was increased after 28 days (p < 0.02). The effect of the treatment with Piracetam was especially accentuated in the patients with aphasia (n = 13), whose status showed the most powerful improvement both after 10 (p < 0.03) and 28 days (p < 0.02). The author believes that Piracetam instituted within the first hours after stroke might improve the neurological status of the patients, especially those with aphasia. PMID- 9748887 TI - Self--administered cough cardiopulmonary resuscitation (c-CPR) in patients threatened by MAS events of cardiovascular origin. AB - Sudden cardiac death caused by cardiac arrhythmia's or asystolies in patients with coronary heart disease can often be avoided if resuscitation is administered immediately, preferably before the patient loses consciousness. In cases when rapid help is not available usually death occurs. We have studied a method of cardiopulmonary resuscitation (CPR) which can be self--administered by patients trained in recognizing imminent arrival of life-threatening Morgani Adams Stokes (MAS) events. The recent study comprised the three methods of investigation in three separate groups of patients: the first group underwent invasive procedures (20 pts), the second non invasive Doppler studies (31 pts) and the third in-and outhospital clinical observations (115 pts). The results indicate that evoked coughing can effectively prevent fainting and maintaining consciousness until conventional CPR help becomes available. PMID- 9748888 TI - [Acute renal failure during pregnancy, labor and puerperium in women]. AB - The reasons and course of acute renal failure during pregnancy, labour and puerperium were presented in 30 women treated in Provincial Hospital in Kielce. The most frequent reason was haemorrhage--15 (50%) women, sepsis--10 (33,3%) women and preeclampsia--eclampsia--2(6, 6%) women. 15 women died as the consequence of multiple organ failure. Among 15 women who survived renal function has returned completely in 13 (86, 6%) ones. In remaining 2 (13, 4%) women chronic renal failure persisted. PMID- 9748889 TI - [The evaluation of action of anesthetic drugs: Xylodont and Mepidont]. AB - The aim of the study was to establish the quality of the therapeutical action- of two local anaesthetics Xylodont and Mepidont, produced by MOLTENI pharmaceutical firm. The investigation was performed in 199 patients of our clinic undergoing teeth extractions or other small surgical procedures. The following data were specified in a questionnaire: name of a drug, applied doses, range of anaesthetic effect, kind and duration of a surgical procedure. We also paid the very attention to the systemic side effects of the used drugs. The significant number of surgical procedures (96, 5%) were painless. The application of additional drug was necessary only in few cases. The obtained results show that the tested anaesthetics Xylodont and Mepidont are safe and effectively acting drugs suitable for local anaesthesia. PMID- 9748890 TI - [The evaluation of the psychosomatic development in children after infectious and idiopathic cholestatic jaundice in neonatal and infantile period]. AB - We carried out the analysis of psychomotoric and somatic development of 38 children after the period of 6--12 months since suffering from neonatal and infantile jaundice (infectious and idiopathic). We did not confirm retardation of the psychomotoric development in the examined group, however we confirmed deficiency of body mass in 45% of patients. Cholestatic jaundice in this group did not influence the children's. psychomotoric development significantly in the following years. It seems, that deficiency of the body mass of examined children after cholestatic jaundice is mostly dependent on the gestational and labour risk factors or small birth weight rather than on clinical course of disease and etiological factors of jaundice. PMID- 9748891 TI - [Liver and endocrine system. Part II: pituitary-gonadal axis and adrenal gland activity disturbances]. AB - Up to date studies have demonstrated that more and more young people are affected by liver diseases. There is a close relationship between the liver and endocrine system as far as hormone inactivation and synthesis of protein binding hormones in the liver are related. Impairment of the hepatocyte function may lead to disturbed homeostasis of the endocrine system. In part II current opinions on plasma levels of pituitary--gonadal and--adrenal axis hormones and their binding proteins in chronic hepatic disorders are presented. PMID- 9748892 TI - [Some aspects of the Clostridium difficile infection]. AB - Clostridium difficile is now regarded as the most common nosocomial enteric pathogen. C. difficile infection has a wide spectrum of a clinical presentation ranging from asymptomatic carriage to the fulminant colitis. Antibiotic therapy is the most important risk factor in pathogen contagion, however other factors are also involved. Typical pathophysiology: 1. alteration of the indigenous colonic flora by antibodies, 2. ingestion of spores, 3. colonization by Clostridium difficile, 4. production of its toxins. Both entherotoxin A and cytotoxin B are active in human colon. The mode of action of these toxins is already quite well known. The main treatment includes withdrawal of the inducing agents, supported occasionally by oral Vancomycin and Metronidazole. Relapse is a major complication. PMID- 9748893 TI - [Large granular lymphocytic leukemia]. AB - Large Granular Lymphocyte (LGL) Leukaemia is uncommon proliferative disorder of LGL lymphocytes. Classification, clinical features and treatment of LGL leukaemia are described. PMID- 9748894 TI - [The lack of parental acceptance of the stress associated with neoplastic disease of the child as a cause of treatment failure]. AB - Three unsuccessful cases of neoplastic diseases in children were described. No parents cooperation with treatment center led after good initial response to the progression of disease and the death of children. The reason of such parents' decisions was incapability to manage the stress caused by child's disease. PMID- 9748895 TI - [Multi-organ tuberculosis in a child: diagnosis and therapeutic problems]. AB - Diagnostic and therapeutic problems in the course of tuberculosis in a 4 year old boy were described. First symptoms of disease such as specific inflammation of lymph nodes were observed in THE third month of liFe. After typical antituberculous treatment the signs of disease regressed. In consequence of the contact with an individual expeCtorating tubercle bacilli superinfection and generalization of the disease took place what led to the boy's death. PMID- 9748896 TI - [Acute cerebellar ataxia in a 5-year-old boy. Clinical warnings]. AB - A case of acute cerebellar ataxia caused by ECHO virus 30.5-year-old boy admitted to the Clinic of Gastroenterology and Nutrition in Warsaw, in September, 1996, complaining of headache, dizziness, weakness, somnolence, dysarthria and an unsteady walk. On neurological examination he had imparied coordination, rombergism, generalized hypotonia. There was no history of exposure to contagious diseases, ear discharge, convulsions, trauma. Parents suggested that the child could have swallowed an unidentified pill--toxicological tests ruled out poisoning. The diagnosis is based on the clinical examination and amplification ECHO virus from CSF. PMID- 9748897 TI - [The traumatic amputation of the forearm complicated by pneumocephalus]. AB - The case of 37-year old man is reported who had traumatic amputation of the right forearm together with brachial plexus and cervical roots and third right rib fracture without pneumothorax. Due to a large damage of the limb replantation was dropped and only surgical elaboration of the wound was made. A few hours after the operation, dyspnoe and headache appeared. New x-ray picture showed right pneumothorax and pneumocephalus with continuing air shadow in cervical vertebral canal. In our opinion the air from pleura might go through cervical intramuscular space to the holes in dura after avulsed roots causing pneumocephalus. PMID- 9748898 TI - [Report on a conference on "Breast carcinoma: prevention, early detection and treatment"]. PMID- 9748900 TI - "I see ads that promote Ensure and Sustacal to older adults. I'm 65; are these supplements worthwhile"? PMID- 9748899 TI - Nutrition. The neglected, nourishing bean. PMID- 9748901 TI - Low back pain. Exercising options for a healthy back. PMID- 9748902 TI - Atrial fibrillation. When a fluttering heart needs attention. PMID- 9748903 TI - Depression and physical decline. PMID- 9748904 TI - Lowering cholesterol. PMID- 9748905 TI - Feasibility of a nurse staffed lift team. AB - The present investigation evaluated the feasibility of creating a lift team with existent medical center nursing staff. Four nurse volunteers at low risk for injury were trained in proper lifting techniques and equipment use. Scheduled and paged maximal assistance transfers were performed by rotating pairs of team members over 21 days. The team successfully completed 94% of requested lifts, averaging 25 per day. Transfers between bed and chair were performed most frequently with the aid of a transfer belt. Greatest demands for team services were on a nursing home unit and an extended/subacute care unit. Unit staff responded positively to the team. Further research is needed to determine the team's impact on unit wide staff injuries and to evaluate the long term effects of working as a lift team member. PMID- 9748906 TI - An effective CPR home learning system. A program evaluation. AB - 1. Program evaluation using a self instruction CPR home learning system indicated that people could learn CPR without attending a formal class. Reported training time for adult CPR averaged 1 hour, and ease of understanding was rated 4.9 on a 5 point scale. 2. Use of a home learning system can benefit a workplace by reducing reliance on a CPR instructor, reducing total training time, and exposing families and others in the community to the learning experience. 3. This program evaluation suggests that this training methodology supports the mission of AHA, and also can meet corporate objectives. PMID- 9748907 TI - Preventing upper extremity cumulative trauma disorders. An approach to employee wellness. AB - 1. Employee education and training about upper extremity anatomy/physiology, risk factors and intervention strategies, including ergonomics, may promote wellness and prevent injuries in the workplace. 2. It is important that nurses in the workplace be proactive in the development and implementation of injury prevention and education programs related to CTDs. 3. Reducing known risk factors and educating workers could have financial benefit for management and should result in healthier, happier employees. PMID- 9748908 TI - Creating constructive outcomes in conflict. AB - 1. Conflict and disagreement are a fact of business life. Effort toward optomizing differences rather than minimizing them is a value added activity- leading to greater creativity, increasing levels of respect in relationships, and better solutions. 2. Proactively looking at potential conflict--where diasgreeing parties are often inherent and/or predictable--can save energy, relationships, and costly mistakes. Diagnosing or "reading" a situation and planning an approach is wise. 3. Several options or responses are available when facing conflict. Knowing when to use a given response is an important interpersonal skill. Relying on learned, habitual, and exclusive approaches to conflict may be limiting. 4. Implementation of effective conflict resolution is a function of attitude, initiative, and flexibility. An exploratory posture and a willingness to learn are constructive in attempting to reach agreements with optimum short and long term effect. PMID- 9748909 TI - Charles Jeffress talks about his priorities for OSHA in the 21st century. Interview by Eileen Lukes. PMID- 9748911 TI - AAOHN Advisory: automatic external defibrillator intervention program. PMID- 9748910 TI - AAOHN Position Statement: use of automatic external defibrillators. PMID- 9748912 TI - Tinnitus as an early indicator of permanent hearing loss. A 15 year longitudinal study of noise exposed workers. AB - A retrospective study was designed to evaluate tinnitus (ringing or other sounds in the ears or head) as a potential early indicator of permanent hearing loss in a population of noise exposed workers. Data were examined from 91 male employees working in environments with noise levels ranging from 8 hour time weighted averages of 85 to 101 dBA over a period of 15 years. Results of annual audiometric testing were obtained as part of an ongoing hearing conservation program conducted since 1971 by ESCO Corporation, a steel foundry located in the Portland, Oregon metropolitan area. Results indicate the prevalence of tinnitus increases more than two and one half times for workers experiencing maximum threshold shifts > or = 15 decibels in hearing level (dBHL). Results also provide evidence that reports of tinnitus at the time of annual audiometric testing may be useful in identifying workers at greater risk for developing significant shifts in hearing thresholds. PMID- 9748913 TI - Medical surveillance of the lead exposed worker. Current guidelines. AB - 1. The "lead standards" established by OSHA for general industry in 1978 and the construction industry in 1993 require employers and clinicians to follow very specific guidelines for protecting lead exposed workers. Depending on the level of exposure, medical surveillance may be legally required. 2. Lead affects multiple body systems and can cause permanent damage. Low level exposures that in the past were thought safe are now considered hazardous as new information emerges about the toxicity of lead. 3. Lead poisoning, if undetected, often results in misdiagnosis and costly care. Adults are exposed to lead in many different workplace settings. All clinicians caring for lead exposed workers need to be informed about the health effects of lead, employer and physician responsibilities, and worker rights. 4. Occupational and environmental health nurses can help identify workers at risk and prevent lead poisoning by education and early intervention through collaboration with the workers, the employer, the company physician, and other health and safety professionals. PMID- 9748914 TI - Hearing conservation. Community outreach program for high school students. AB - 1. A community service project for a local constituency of the AAOHN involved teaching hearing conservation classes in junior high school shop classes. 2. Pre test/post-test comparisons showed that students benefited from the 1 hour class, including indications they planned to use hearing protection devices in the presence of loud noise. 3. Occupational health nurses' expertise in the area of hearing conservation can enhance work-school partnerships that strengthen communities. PMID- 9748915 TI - Domestic violence in the workplace. AB - 1. It is important for workers, management, and health and safety personnel to recognize a history of domestic violence may place an individual at risk for acts of violence at work. 2. Employers and unions are beginning to recognize domestic violence may impact the economic well being of a workplace by decreasing productivity and attendance, and by increasing insurance and health care costs. 3. Few companies have policies related to general safety, general workplace violence, or specifically to domestic violence. All departments within the corporation or business should help to develop and implement these policies, and training in the policies must include all levels of the business. PMID- 9748916 TI - HIV litigation update. PMID- 9748917 TI - Social role quality and psychological well being in employed black and white midlife women. AB - This study examined relationships among job, partner, and parent role quality and psychological well being in midlife black (n = 51) and white (n = 56) women employed in occupations varying by socioeconomic status (SES). Oversampling for black women ensured balanced occupational representation, allowing investigation uncontaminated by SES. Instruments included Baruch and Barnett's Rewards and Concerns Scales, Bradburn Affect Balance Scale, and Center for Epidemiological Studies Depression Scale (CES-D). Better well being scores were reported by black women than whites, and by professionals than non-professionals. However, when race, occupational group, and menopausal status were held constant in a multiple regression analysis, partner role quality was significantly related to both well being scores, parent role quality was related to life satisfaction only, and job role quality was not related to either. Nurses in the workplace can help women identify problematic aspects of their multiple social roles, and facilitate resolution of problems to improve worker health. PMID- 9748918 TI - Prostate cancer screening in the workplace. Employer costs. AB - Recognition of the mortality and morbidity associated with prostate cancer has resulted in employer based screening programs. This retrospective cohort study identified the employer costs of prostate cancer screening and referrals due to abnormal test results. The subjects were 385 men enrolled in a workplace screening program at a single employer between 1993 and 1995. Screening consisted of digital rectal examination (DRE) annually for enrolled employees aged 40 years and older, plus annual prostate specific antigen (PSA) testing for those 50 and older, and those 40 and older and considered at high risk. Data related to the health care and lost productivity costs of screening and referrals for abnormal test results were collected and analyzed. The total cost of screening was $44,355, or approximately $56 per screening encounter (788 DREs; 437 PSAs). Abnormal screening tests resulted in 52 referrals. Upon further evaluation, 42% were found to have an enlargement, 29% a node, and 12% benign prostatic hyperplasia. Only one malignancy was found. The total cost of additional referrals was $31,815, or 42% of the cost of screening plus referrals. As the cost per screening encounter was low, prostate cancer screening in the workplace is an efficient alternative. PMID- 9748919 TI - Linking resources to process in disability management. Successful program. AB - 1. Occupational health services need to link resources to business process to product to support the business goals of the corporate organization. 2. Use of business process steps in business plan development provides an organized, comprehensive approach to effective product development. 3. It is essential to demonstrate a cost reduction which is reflected in the business organization's ROI, resulting in the shareholder's increased per share earnings. PMID- 9748920 TI - Project management skills. AB - 1. Project management skills are important to develop because occupational and environmental health nurses are increasingly asked to implement and manage health related projects and programs. 2. Project management is the process of planning and managing project tasks and resources, and communicating the progress and results. This requires the coordination of time, tasks, equipment, people, and budget. 3. Three main critical skill areas are needed to be an effective project manager: behavioral skills such as negotiation, conflict resolution, and interpersonal problem solving; use of project management tools to manage project tasks and resources; and effective communication skills. PMID- 9748921 TI - Writing effective reports. PMID- 9748922 TI - Finding information on the Internet. PMID- 9748923 TI - Research award: PCNPs and occupational illness or injury. PMID- 9748924 TI - Nursing older people: learning from experience. PMID- 9748925 TI - Outcomes steering practice: when the ends determine the means. AB - A discussion of outcomes and outcomes measurement will be presented through an examination of achievements of a collaborative project awarded a federal best practice grant in 1995. The project, which established a new service for women of nonEnglish-speaking background, was part of a national venture which sought to benchmark best practice in the primary health care field. The process of benchmarking demanded that the approach to outcomes and outcomes measurement adopted by the project participants be articulated. The approach adopted acknowledges that outcomes are affected by the processes involved in care, which in turn are dependent upon the organisational environment in which care is offered and experienced. Both these contributing factors to outcomes can be hard to measure and, therefore, could be at risk of being omitted from the outcome equation in these economically stringent times. This paper aims to contribute to debate on the topic by recommending an approach to outcomes measurement which is comprehensive, values quality and embraces the short, medium and longer term. PMID- 9748926 TI - Seeking comfort through prayer. AB - The aim of the research was to discover the experience and meaning of prayer of patients in hospital when undergoing coronary artery bypass graft surgery. Ethnographic-type post discharge interviews with 13 participants formed the major data source. Using the grounded theory method, the basic social psychological process was labeled seeking comfort through prayer. This was the process of engaging in prayer with God. The participants believed that God listened to their prayers and answered them when they were seeking comfort. It was this reassurance that gave them the strength to face uncertainty and possible death, and also gave them comfort in their psychophysiological condition at the time. Seeking comfort had three stages (maintaining or re-establishing a relationship with God; making peace with God; and asking God to be with them during the hospitalisation). This last stage involved five levels of prayer (acquiescence, instinctive prayer, survival, confiding and honouring). PMID- 9748927 TI - Elderly Finnish people's experiences with coping at home. AB - The aim of this study was to describe the experiences of elderly Finnish people with coping at home. Twenty elderly inhabitants of the city of Oulu in Finland, over 75 years of age, who live at home were interviewed. Content analysis was used as the method. According to the analysis, elderly people's coping at home consisted of social contacts, daily events as the substance of life, and previous life experiences. Social contacts consisted of family, public health services, and neighbours. Daily events as the substance of life included taking part in activities of daily living. The contents of previous life experiences consisted of thinking back to organize one's life experiences and their influence on one's own life. The factors that promoted coping at home were maintenance of health, the experience of well-being, and security. The findings of this study indicate that by supporting elderly people to cope at home, it is possible to influence their sense of safety and well-being and hence their quality of life. PMID- 9748928 TI - Treating skin tears in nursing home residents: a pilot study comparing four types of dressings. AB - A pilot study was conducted to compare four types of dressings used to treat skin tears in nursing home residents. Wounds treated with a non-occlusive dressing healed more quickly than those dressed with occlusive dressings. The results suggest that ease of use and product wastage are important considerations when treating skin tears. The pilot study also highlights the need for further research into skin tear management and the need for ongoing education for nurses regarding skin integrity risk assessment and product information. PMID- 9748929 TI - A descriptive study examining postdischarge patient needs after laser ablation and transuretheral resection of the prostate. AB - This descriptive study identifies and describes the postdischarge experiences in two groups of patients with benign prostate hyperplasia. The two patient groups included 12 patients who underwent laser ablation and 12 patients who underwent traditional transuretheral prostatectomy. A questionnaire was developed and used to examine the postoperative experiences of the two groups after discharge. The questionnaire obtained demographic information from medical records and examined the types of assistance required at home, the need to contact health professionals, difficulties encountered after catheter removal and the amount of time until resolution of symptoms and a return to normal activities. The laser ablation group were discharged home with an indwelling catheter, and thus were required to complete additional questions regarding their experiences of going home with a catheter and leg bag. The results indicate the laser ablation group went home with an indwelling catheter and reported a higher frequency of symptom distress and technical problems in caring for the leg bag. The laser ablation group also sought more assistance from health professionals. These problems were not demonstrated in the traditional transuretheral prostatectomy group, who remained in hospital until the catheter was removed. This study demonstrates the need for further nursing research examining patient issues that occur during the postdischarge recovery phase. PMID- 9748930 TI - School nurse assessment of primary school children: analysis of data from the school entrant health questionnaire. AB - The School Entrant Health Questionnaire (SEHQ) has been used by the School Nursing Program in Victoria since April, 1997. The SEHQ assists school nurses in developing a health profile of primary grade children and in discriminating between children with problems and children without problems. The SEHQ assesses children in 11 domains: general health, medications, immunisations, dental health, speech/language, hearing, vision, disabilities, general development, behaviour and emotional well-being, and family stress. The SEHQ was found to be reliable and valid, and to provide an excellent means of distinguishing between students who had problems and needed intervention and those who did not. This paper presents an analysis of data from the first testing of the SEHQ. PMID- 9748931 TI - Restoring homeostasis in a residential care facility through behaviour modification. AB - A verbally aggressive, 32-year-old male with a traumatic brain injury was admitted to a unit in an aged care facility for residential care. The homeostasis of the unit was disrupted by the resident's verbal aggression and other inappropriate behaviours. With the guidance of a neuropsychologist, nursing staff were able to use behaviour modification to successfully replace the disruptive behaviours with more socially appropriate ones. A series of positive rewards was implemented in response to socially appropriate behaviour, whilst inappropriate behaviours received a negative reward. Several disruptive behaviours were affected by the single treatment implemented. This interdependence of targeted behaviours was viewed as a clinical advantage, as it served to provide a more rapid restoration of homeostasis to the unit. The use of a single-subject, multiple baseline design in this case study demonstrates that disruptive behaviours may be reversible. PMID- 9748932 TI - They don't pay you girls enough. PMID- 9748933 TI - To live, not only to survive. PMID- 9748934 TI - United Kingdom: caring differently: intermediate care--an alternative approach to service provision in response to client need and workforce changes in the NHS. PMID- 9748935 TI - The rhythm of life. PMID- 9748936 TI - New approaches to health and well-being for dementia day-care clients, family carers and day-care staff. AB - This study was conducted in one multicultural dementia day-care centre over a period of 18 months. It introduced a gentle hand treatment for clients using three essential oils. The study evolved out of the process of action research where the family carers and day-care staff participated with the researchers to choose, design, develop and evaluate a hand treatment programme. Data was collected through in-depth interviews pre- and post-treatment, focus group discussions, client observation logbooks and a disability scale. The findings indicate a positive strengthening of the relationship between the person with dementia and their family carer, and an improvement in feelings of health and well-being for both. The specific improvements for clients include increased alertness, self-hygiene, contentment, initiation of toileting, sleeping at night and reduced levels of agitation, withdrawal and wandering. Family carers have reported less distress, improved sleeping patterns and feelings of calm. They also found the treatment useful in helping them manage the difficult behaviours exhibited by their relative with dementia. The benefits of this treatment for nursing practice are that it is safe, effective and easily administered by staff in any setting. PMID- 9748937 TI - Analysis of the stressful effects of hospitalisation and source isolation on coping and psychological constructs. AB - This quantitative research has attempted to investigate the psychological effects of hospitalisation and source isolation, and assessed whether were they influential in affecting a patient's cognitive coping with these two stressors. The research evaluated whether isolating a person because of an infection was a more stressful event (causing negative effects on four measured psychological constructs) than routine hospital admission. The research was conducted in two large District General Hospitals and one elderly care hospital. Individuals admitted to one of the research sites, and who satisfied the sample criteria, were adopted. The total number of subjects was 40. The research design was quasi experimental (post test only control group design), using a quantitative approach. Following a period of hospitalisation or isolation subjects in the control group (Group 1, hospitalised subjects n = 20) and subjects in the experimental group (Group 2, isolated subjects, n = 20) were given the following to complete: the Hospital Anxiety and Depression Scale, the Health Illness (Powerlessness) Questionnaire, and the Self Esteem Scale. These measured four psychological constructs: anxiety, depression, self esteem and sense of control. The quantitative data generated were analysed using descriptive statistics and the Student's t-test. The findings confirmed and validated previous research that hospitalisation results in many negative feelings that have detrimental effects on psychological well being and coping. However, more significantly, infected subjects who were isolated demonstrated feelings of anxiety, and depression that were significantly higher, and feelings of self esteem and sense of control that were significantly lower than those demonstrated by hospitalised subjects. Thus it could be argued that isolation has an even greater negative effect on their coping. Further research therefore needs to examine how specific nurse interventions can ameliorate the identified negative effects of isolation and so facilitate effective coping and positive psychological well being. PMID- 9748938 TI - Nurses' views on reporting medication incidents. AB - The purpose of this project was to identify nurses' beliefs about medication incident reporting. A new medication incident form was developed and trialled in six clinical units. Forty-three nurses from these areas were recruited to participate in the project, with a 20-point self-reporting questionnaire and focus group discussions being used to collect the data. Theme analysis of the data was undertaken with the results of the project indicating nurses report medication incidents that are life threatening to patients, but do not want identifying information collected about themselves. This situation represents nurses' fear of reprimand from those in authority and may also indicate an unwillingness to accept responsibility for errors in which they may be merely the final player in a complex series of events. The results of the project also highlight problems associated with self-reported medication incident monitoring and challenge its effectiveness in gathering data required by managers and staff development educators. PMID- 9748939 TI - Life control and health in view of qualitative and quantitative research. AB - In this paper the experiences of life control and health of Finnish people are described and compared by reference to two studies with different methodologies. The goal of the qualitative study was to describe and understand the human being's health as an individual way of existence on the basis of life descriptions. The qualitative data were gathered through thematic interviews (n = 60, men and women, aged 30-50 years). The data were analyzed with the grounded theory method. The analysis showed life control to be a core category that characterizes health experiences. On the basis of the analysis, the content of concept of life control was defined to describe the data. The goal of the quantitative study was to produce information on life control among young males, their health, health behaviour, experiences of stress and life situations as well as the associations between these dimensions. The definition of life control was based on Antonovsky's theory of sense of coherence and on the study of Soderqvist and Backman. The quantitative data were gathered with questionnaires from 2500 (response rate 60%). The data were processed using cross-tabulation and multivariate data analysis (discriminant analysis). The connections between life control and health were obvious in both studies. To a certain extent the results were parallel, but there were also some differences which are discussed here as a starting point for developing further the research on life control and health in the nursing science context. PMID- 9748940 TI - Meaning-making for family carers in nursing homes. AB - This ontological hermeneutic study highlights the importance of understanding the human experience of family caring. In contrast to much family caregiving research which focuses upon the home care situation, this study involved in-depth audiotaped conversational interviews and observations with 14 family carers who continue to care within a nursing home context. Thematic analysis of the transcribed interviews/field notes uncovered a number of common themes of meaning which highlight the nature of family caring experience in nursing homes. Discussion of such findings will challenge practitioners to reconceptualise the nurse resident-family carer relationship, appreciate the many ways in which a family member's involvement in care provides meaning and significance in their lives, and understand family carers through a process of human relating which fosters families' meaningful involvement in caring within a nursing home context. PMID- 9748941 TI - Creating a nursing development unit in a dementia care context. AB - This paper describes the process through which a Nursing Development Unit (NDU) was created in a 29-bed unit in which nurses care for severely demented residents. The question about whether or not ethics clearance is required for the development phase of NDUs is discussed, and the framework in which to develop the NDU described. The benefits of using Participatory Action Research as the basis for undertaking a continuous process of reflection and change is highlighted. It is also suggested that two key elements necessary for NDUs are that they are 'owned' by the staff who work in them and that they are supported by senior management of the organisations in which they exist. PMID- 9748942 TI - Patient aggression in a general hospital setting: do nurses perceive it to be a problem? AB - Aggressive incidents in the general hospital setting have been recognised as problematic for health care workers. Despite an awareness of the problem by nursing staff, there is little known about these incidents. A survey was therefore conducted in a metropolitan tertiary hospital to determine the prevalence and extent of patient aggression in order to direct future management strategies. A survey tool elicited information about the aggressor, factors leading to the incident, the nature of the incident, how it was managed and the outcome. Sixty-eight incidents of aggression were reported over a five-month period; all were reported by nursing staff. The majority of incidents occurred after hours when staff resources were limited. Frequent actions taken by staff to manage aggressive patients were chemical and physical restraint. Nurses identified that many of the incidents were unavoidable despite the aggression management training they had received. This paper outlines strategies taken by the hospital to address the issues identified in the survey. PMID- 9748943 TI - Reviews of units of care: combining practice, research and quality assurance. AB - That nurses should base their practice on the best available knowledge is generally accepted by the profession. However, the operationalisation of this ideal if difficult for hard pressed clinicians. We believe that responsibility for assimilating current knowledge in a practical format belongs to all nurses working in an organisation. This paper describes a project where clinicians, administrators and academics have collaborated to rewrite standards of practice in a format that includes up to date evidence and practical measures for appraising outcomes. This work is a practical example of quality assurance activities serving to create opportunities for collaboration between nurses who have chosen to practice, teach, manage and/or research nursing. PMID- 9748944 TI - [Nutrition of premature infants and of infants with low birth weight with mother's milk or donated human milk]. PMID- 9748945 TI - [Nutrition during the first days of life]. PMID- 9748946 TI - [Illustrations of nosocomial pediatric infections]. PMID- 9748947 TI - [Premature infants and their parents--integration of parents into the neonatal intensive care unit]. PMID- 9748948 TI - [Lightning out of the blue sky?--what one should know about epilepsy]. PMID- 9748949 TI - [When religious problems come up during conversations with parents]. PMID- 9748950 TI - [Pediatric nursing at home. The care of sick children by organizations for home pediatric care]. PMID- 9748951 TI - [Patient care planning--one more thing]. PMID- 9748952 TI - [Legislation for the protection of mothers. The employer has the right to demand an investigation of an occupational prohibition]. PMID- 9748953 TI - [We shall fashion our own future]. PMID- 9748954 TI - [Delegates meeting of the Swiss Nursing Association. Opposing the pressure]. PMID- 9748956 TI - [Everybody has inalienable rights]. PMID- 9748958 TI - [As the patient is positioned...]. PMID- 9748957 TI - [Employment on call: unacceptable]. PMID- 9748959 TI - [A little known element of modern ulcer therapy: moist therapy of chronic skin ulcers]. PMID- 9748961 TI - [Decubitus ulcer. An unusual plan for prevention. Where the shoe hurts]. PMID- 9748962 TI - [Decisions concerning the future]. PMID- 9748965 TI - [A magic chariot in the hospital]. PMID- 9748966 TI - [Treatment of kidney failure with peritoneal dialysis: an interesting alternative]. PMID- 9748967 TI - [For me peritoneal dialysis is a solution while waiting for a new kidney. Interview by Brigitte Kocher-Longrich]. PMID- 9748968 TI - [Quality intensive care. Does psychiatry need humaneness?]. PMID- 9748969 TI - [The Lausanne College of Nursing Education. New perspectives]. PMID- 9748970 TI - [Passion and compassion in nursing]. PMID- 9748971 TI - Coping with 'slippery slope' questions. PMID- 9748973 TI - The history and impact of worksite wellness. AB - Employers now pay an estimated 30% of the national health care bill. Wellness is defined as "a composite of physical, emotional, spiritual, intellectual, occupational, and social health; health promotion is the means to achieve wellness." Worksite wellness programs have developed largely in response to cost containment efforts combined with the worksite health promotion movement. Worksite wellness efforts require the use of a model that targets reversible or alterable behaviors such as smoking, weight management, blood pressure monitoring, and stress management. Most employers in the health care arena are seen as doing too little to promote wellness among its own employees. PMID- 9748972 TI - Finding value in nursing care: a framework for quality improvement and clinical evaluation. AB - Fiscal constraints have heightened attention to health care costs and patient outcomes as measures of health care system effectiveness. Determining which patient and costs outcomes nurses may be held accountable for requires differentiating the impact of dependent, independent and interdependent nursing activities. A nursing role effectiveness model that includes a number of structural variables is offered to help track quality improvement and research activities. Some of the nurse-sensitive patient outcomes that have been identified include: freedom from complications, clinical outcomes, functional health outcomes, knowledge outcomes, perceived health benefit (or satisfaction), and costs outcomes. This model can be used to evaluate the effectiveness of current as well as evolving nurse roles, processes, and structural changes. PMID- 9748974 TI - A point of view: why point-of-care places are not free marketplaces. AB - Current wisdom holds that health care is a business and "as such must abide by market principles." Most nurses are not well enough versed in economic theories to credibly critique health care delivery decisions based on economic theories. The relationship of market principles to health care realities is described in basic terms to encourage nurses to "optimize patient care and influence health care policy." Physicians, who control all access points to the health care system, have enjoyed a 40-year market dominance that is "rapidly being replaced by insurance companies and for-profit investors." Providers' decisions to treat or not to treat are strongly influenced by whether the patient is in a fee-for service or capitated payment environment. PMID- 9748975 TI - Basic statistics for clinical pathway evaluation. AB - Clinical pathways (using accepted benchmark goals) are among the most widespread tools used to enhance outcomes and contain costs within a constrained length of stay (LOS). The author describes the need for selection and use of designated basic statistics in the "definition and evaluation of the impact of clinical pathways in hospitals." The use of both the mean and median data on resource use and LOS information (to evaluate the effectiveness of various clinical pathways) is advised, as the influence of outlier data will thus be revealed. The ultimate goal of such evaluation is the identification of statistics that can be readily "understood by and communicated among providers and consumers of health care services." The correlation of statistical variables such as resource utilization (including ICU stays) and LOS, can reveal patterns not easily seen otherwise. PMID- 9748976 TI - Interviewing job candidates' references: a key hiring strategy. PMID- 9748977 TI - Gleaning patterns from practice: intelligent systems in ambulatory care. PMID- 9748978 TI - Understanding the Y2K problem in health care: party's over, we're out of time. PMID- 9748979 TI - Health care fraud: hemorrhage from the health care system. PMID- 9748980 TI - Management is taught, leadership is learned. AB - Management can be taught. Leadership must be experienced to be learned. The great leaders have evolved their abilities through on-the-job experiences that prepare them to lead by a variety of successes and failures. The quicker you get started on this journey to leadership, the more time you will have to build a legacy of positive outcomes. The patient care unit manager is a very valuable person. However, we need many more leaders who can think out of traditional boxes, lead people enthusiastically into the new world of health care, and transform our profession and the health care system to more effectively meet the health needs of our society. The best we can do is to empower people to start this journey of leadership early and seriously in their careers. PMID- 9748981 TI - On measuring and managing ... PMID- 9748983 TI - Patient-focused care: what managers should know. AB - Implementation of PFC is considered a "rational" strategic choice in that it is thought to decrease the cost of providing health care while increasing the quality of services. Published research and evaluation studies that describe PFC are analyzed and the information is synthesized in hopes of discovering and describing commonalties among definitions, goals, and principles underlying PFC. The commonly accepted description of PFC is a model which "seeks to integrate the organization's values and culture with the operational excellence vision and processes to transform the institution into a customer-focused organization." Staff satisfaction is addressed by encouraging staff to plan and execute their clinical work in ways that are most responsive to patient needs. Grouping similar patients, bringing services closer to these patients, and appropriate cross training of multidisciplinary care providers to enhance continuity of care are seen as the four most common elements described in most of the literature. PMID- 9748982 TI - Forecasting the nursing workforce in a dynamic health care market. AB - The ability to discern the interacting factors that affect supply and demand for nurses could help nurse educators and nurse leaders allocate resources to meet these needs. Forecasting models must take into account the interactions of three crucial groups of health care providers--physicians, nurse practitioners, and physician's assistants. Buerhaus has noted that market size, wages, preferences for nursing services, and availability of substitutes influence the demand for nursing services. Changes in nurse supply resulting from Medicare reimbursement for nursing services have not been studied, though it could safely be projected that such reimbursement will increase nurse supply. Nurses with baccalaureate degrees and advanced practice preparation will be in the greatest demand in ambulatory care, managed care, public health, and home care settings, raising concerns again that the educational mix is in need of adjustment upwards. PMID- 9748984 TI - Adult daycare: an entrepreneurial opportunity for nursing. AB - By the year 2000 13% of the general U.S. population will be made up of those over 65. Many states are seeking cost-effective ways to provide structure and some assistance for these individuals to help keep them in their own homes and out of institutions. The National Institute on Adult Daycare (NIAD) defines adult daycare as "a community-based program designed to meet the needs of minimally impaired adults through an individualized plan of care for part of a 24-hour day." Between 1980 and 1990 the number of centers grew from 1,200 to 3,000; by the year 2000, NIAD estimates that there will be a need for 10,000 adult daycare centers. The authors advise interested nurse entrepreneurs to consider the potential for establishing and running such centers and provide a roadmap for developing a business plan. A careful market analysis in your selected community should be the first step in the process of developing a workable business plan. PMID- 9748986 TI - What is your legacy? PMID- 9748985 TI - A multisite study of nurse staffing and patient occurrences. AB - Restructuring of nursing care models has led to more "non-professional" caregivers, sometimes called unlicensed assistive personnel (UAPs) who provide more of the basic delegable direct patient care activities in collaboration with RNs. The purpose of this study, wherein data were collected from 39 units in 11 hospitals, was to determine the relationship between different levels of nurse staffing and patient outcomes (adverse occurrences). Using and tracking the same indicators of patient quality outcomes over a significant time period in different institutions with similar patient groups would greatly enhance the usefulness of such data. Among the more surprising findings in this study was the "non-linear" relationship between the proportion of RNs in the staff mix and MAEs. As the proportion of RNs on a unit increased from 50% to 85% "the rate of MAEs declined, but as the RN proportion increased from 85% to 100% the rate of MAEs increased." Further investigations are needed to explain this finding. PMID- 9748987 TI - Fighting the cold war: how information technology can help. PMID- 9748988 TI - Update on patient privacy legislation. AB - The administrative simplification provisions of HIPAA will establish the first national standards for the electronic transmission of health care transactions with which all federal programs (DOD, Medicare, and Medicaid) and all private health plans must comply. Individuals and organizations should prepare themselves, their systems, and their processes to meet these new administrative and financial data standards and requirements. The benefits of standardized electronic transactions on achieving a single paper-free claims submission to be used by all providers and payers is of obvious benefit. Not so obvious are consequences associated with limiting the access and use of existing data repositories on a variety of clinical, administrative, and research functions. It is critically important in this age of increased accountability for fiscal restraint and improving the outcomes of entire patient populations that clinicians, managers, organizations, and researchers to use data for a variety of clinical, quality improvement/evaluation, and research purposes. Administrative simplification and protecting individual privacy should not be achieved by overly bureaucratic and restrictive responses that impede epidemiologic and health services research, quality improvement activities, and optimization strategies for improving the health of populations. While the health system understands the need for some increased regulation to ensure the privacy of individual patient privacy in the "wired" world solutions must be found and overly restrictive consequences associated with prohibiting access to data must be resolved. More than ever, the entire system requires data to inform every level and type of decision made. Legislation and bureaucratic processes that do not understand and support responsible data-driven decision-making will serve to roll-back, not advance health system improvement. As we prepare ourselves for HIPAA compliance and the expectations of the benefits it will achieve, we must wait to see what impact it will have on clinical, administrative, and research functions concerned with improvement. PMID- 9748989 TI - Growth strategies to optimize the functions of telephonic nursing call centers. AB - The changes occurring in the health care delivery system afford ideal opportunities for call centers to expand their essential functions. Two obvious and timely services that can be adapted to the call center are outcomes management and disease management. These services benefit from the central role that telephonic nurses can play in clinical assessment and data collection and analysis. Other new services, such as gate-keeping functions, may also be relevant to call centers. The information and technology specialization of expert clinicians who practice "sightless" nursing make call centers the new clinical epicenter in the service capabilities of health care networks. PMID- 9748990 TI - The strategic use of humility. AB - Authentic humility is strategic for the success of leaders. It is strategic from the perspective of the egotistically free relationship humility establishes with the staff, patients, and others. The lack of strategic humility explains the inevitable downfall of people who are motivated to lead by power and status. It also explains why leaders who always remember where they came from, and who are motivated by what they can do for patients and staff, succeed. PMID- 9748991 TI - Clinical effectiveness for nurses, midwives and health visitors. PMID- 9748992 TI - Nutrition. PMID- 9748993 TI - Quality time. PMID- 9748994 TI - In search of common ground. Interview by Rebecca Coombes. PMID- 9748995 TI - Will the truth still hurt?. Interview by Caroline White. PMID- 9748997 TI - How should nurses respond to patients who are blind? PMID- 9748996 TI - Ironically it was a terrible night on duty. PMID- 9748998 TI - How to grow more leaders. PMID- 9748999 TI - Learning to fly. PMID- 9749000 TI - United States. Interview by Rob Garbett. PMID- 9749001 TI - A long time coming. PMID- 9749002 TI - Practical procedures for nurses. Last offices--3. PMID- 9749003 TI - In line for promotion. AB - Many nurses are unfamiliar with the issues surrounding public health promotion. Elizabeth Meerabeau reports on what they mean for education and training. PMID- 9749004 TI - Good news down the line. PMID- 9749005 TI - A nurse in agony. PMID- 9749006 TI - Periodic registration is a personal responsibility, but employers are expected to check it. PMID- 9749007 TI - An alternative to inpatient care for clients with acute mental illness. PMID- 9749008 TI - Bereavement support for people with learning disabilities. PMID- 9749009 TI - Research, audit and networking: who's in the lead? AB - This article compares the progress made by nursing development units compared with matched units without NDU status on research, audit and networking activities. Research and dissemination is significantly greater in NDUs than in comparators. Research in NDUs focuses on local attempts to improve practice and has some way to go before it can be generalised to health care settings beyond the unit where it was carried out. There is no difference in number of audits undertaken by NDUs and comparators but NDUs carried out more audits to evaluate nursing work and comparators did proportionately more on practice topics and user issues. There is no association between research and audit activity; more audit does not result in less research Networking activity by staff is significantly greater for NDUs, suggesting they are seen as a resource for other centres. NDUs are active in research and networking--significantly more so than comparators. PMID- 9749010 TI - Dirt alert. PMID- 9749011 TI - Food fears. PMID- 9749012 TI - Parade and prejudice. AB - Nurses at Craigavon hospital near Drumcree braved bomb threats and violence during the Orange parades weekend to carry on with their work. Adrian O'Dowd was invited to join them. PMID- 9749013 TI - Cold comfort. AB - The government's comprehensive spending review gives the NHS a welcome cash boost. But unions say it will backfire without investment in the workforce. Nursing Times reporters gauge the prevailing mood. PMID- 9749014 TI - Fever pitch. Interview by Barbara Millar. PMID- 9749015 TI - The fallout from Viagra means it is time to come clean about NHS rationing. PMID- 9749016 TI - A spin doctor for the NHS. Surely a sign of the times. PMID- 9749017 TI - Some nurse academics are short-sighted about where research results should be published. PMID- 9749018 TI - The government's obsession with spin doctoring--putting the best gloss on any story--is rebounding, causing damage. PMID- 9749019 TI - The future of contraception. PMID- 9749020 TI - Writing the changes for nurses. AB - Prescribing powers for family planning nurses has been a heated and topical issue for decades and progress has been impossibly slow. Jane Urwin wonders if this extended role for nurses will ever materialise. PMID- 9749021 TI - Sex and disability. PMID- 9749022 TI - Teenage sex talk. PMID- 9749023 TI - Practical procedures for nurses. Nutrition assessment. PMID- 9749024 TI - Gilles de la Tourette syndrome. PMID- 9749025 TI - The voice of the people. PMID- 9749026 TI - Partnership in action. AB - Patient participation groups do work. Pat Goodwin explains how she has been able to give something back to the practice where she has been registered for over 15 years. PMID- 9749027 TI - The plague wars. PMID- 9749029 TI - Changes to the ENB's Family Planning Course have made further training impractical and almost impossible for practice nurses. PMID- 9749028 TI - The great leveller. PMID- 9749030 TI - The endocrine system. Hypothyroidism. PMID- 9749031 TI - Assessment of haemostasis. AB - Major surgery and some therapies carry with them the chance of severe bleeding. Recognising who is at risk and knowing what to do if it happens are important parts of the nurse's job. Debbie Field explains. PMID- 9749032 TI - Self-destruction in the pancreas. PMID- 9749033 TI - Self-management in primary care. PMID- 9749034 TI - We can work it out. PMID- 9749035 TI - Helping students find a place in the team. AB - Confusion still surrounds the issue of supernumerary status. Annette Lankshear recommends a pragmatic approach to the education of Project 2000 students in the wards. PMID- 9749037 TI - Age no bar to a caring career. PMID- 9749036 TI - Survey looks at problems in university nursing research. PMID- 9749038 TI - To the House of Lords. PMID- 9749039 TI - The cost of overseas aid. PMID- 9749040 TI - Hail Marie Curie. PMID- 9749041 TI - Education and training on domestic violence is necessary to enable organisations to support their staff and tackle the dangerous myths surrounding it. PMID- 9749042 TI - The disease of deceit. PMID- 9749043 TI - The hospital hoppers. PMID- 9749044 TI - The legacy of Allitt. AB - Since nurse Beverly Allitt's Munchausen's syndrome by proxy led her to kill children in her care, recruiting managers have been wary of mental illness. Now it is time for a more positive approach, say Sheelagh Brewer and Chris Cox. PMID- 9749046 TI - Face value. PMID- 9749045 TI - Help that goes direct to the heart of the matter. PMID- 9749048 TI - Workers' health undermined. PMID- 9749047 TI - Practical procedures for nurses. Catheter specimen of urine. PMID- 9749049 TI - Anchors aweigh. AB - It is appropriate that a ship named after Britain's best-known sailor should give people with disabilities the chance to sail the open seas. Nurse Jane de Burgh helped out on a voyage of self-discovery. PMID- 9749050 TI - Would you credit it? PMID- 9749051 TI - Information for patients having bladder augmentation. AB - Bladder augmentation improves quality of life, but it carries risks. Nurses are in a prime position to ensure patients give fully informed consent before surgery, as Catherine Savage explains. PMID- 9749052 TI - Primary health care for people with learning disabilities. AB - Health screening and education for people with learning disabilities is often given low priority by service providers, but the input of community nurses can make a great difference. Helen Cook charts the success of two teams. PMID- 9749053 TI - Problems and procedures of day-case angiography. PMID- 9749054 TI - Avoiding drug administration errors: the way forward. PMID- 9749055 TI - The medicine wheel and the millennium. AB - Do we place too much importance on rules and protocols in the work environment? The holistic approach argues Alys Harwood, can bring clarity and purpose to management roles. PMID- 9749056 TI - The pitfalls of practice nursing. PMID- 9749057 TI - Great expectations. PMID- 9749060 TI - [The Euro is coming. By 2002 the German mark will be finished]. PMID- 9749058 TI - [The industry wants to intensify dealings with foreign countries]. PMID- 9749061 TI - [The infrastructure for quality assurance is missing]. PMID- 9749062 TI - [50 years of Pflegezeitschrift: nurses are madly writing]. PMID- 9749063 TI - [Nursing as a problem-solving and a relational process: the individual patient's situation has to be considered]. PMID- 9749065 TI - [Educational trip to New York: ... to see how the others are managing]. PMID- 9749064 TI - [The caregiver decides--or does he? 2. Often the guardianship court has to decide]. PMID- 9749066 TI - [System of quality assurance in the hospital: nursing often has the avant garde role of a lone fighter]. PMID- 9749067 TI - [Social dynamics of start and end situations: every ending is the beginning of something new]. PMID- 9749068 TI - [Quality management at the University of Pittsburgh Medical Center Presbyterian Hospital, Pennsylvania. Report on a practical stage]. PMID- 9749069 TI - [Prognostic factors of infiltrating tumors of the bladder]. AB - In France, invasive bladder cancer is the more frequent urologic malignancy after prostate carcinoma. Treatment of bladder cancer is radical cystectomy. New therapeutic approaches such as chemoradiation combination for a conservative procedure, neoadjuvant or adjuvant chemotherapy are still developing. In this way, a rigorous selection of patients is needed. This selection is based on prognostic criteria that could be divided into four groups: i) the volume of the tumor including the tumor infiltration depth, the nodal status, the presence or not of hydronephrosis and the residual tumor mass after transuretral resection; ii) the histologic aspects of the tumor including histologic grading, the presence or not of an epidermoid metaplasia, of in situ carcinoma or of thrombi; iii) the expression of tumor markers (tissue polypeptide antigen, bladder tumor antigen); iv) the biologic aspects of the tumor as ploidy, cytogenetic abnormalities, expression of Ki67, expression of oncogenes or tumor suppressor genes, expression of tumor antigens or growth factor receptors. This paper reviews the prognostic value of the various parameters. PMID- 9749070 TI - [Flow cytometry in cancer of the bladder]. AB - We reassessed the use of DNA flow cytometry in bladder cancers on the basis of our research and already published findings. We discuss technical aspects underlying the validity of the results. Currently, the validity of DNA flow cytometry is established by parametric analysis of the DNA content of tumor cells found in the course of multiple biopsies of the tumor. In addition, we examine the results obtained with bladder washings and, in some cases, the results of biopsies of the bladder mucosa which may appear normal under cystoscopy. The complementarity of these examinations appears to be essential. Our experience confirms the results already published, suggesting that the frequency of DNA aneuploidy increases significantly according to the grade and the tumor stage. However, clinical interpretation of DNA flow cytometry results calls for some caution. There is a general consensus not to use these results in the screening of bladder cancers. However, DNA flow cytometry is particularly useful in the follow-up of carcinoma in situ since DNA aneuploidy is almost always present. DNA flow cytometry is also useful in the stratification of superficial grade 2 tumors. Finally, during the follow-up of invasive tumors, the persistence or appearance of DNA aneuploidy may be attributed to therapeutic resistance. PMID- 9749071 TI - [Therapeutic approach to bladder cancer classified as T3]. PMID- 9749072 TI - [Treatment of cancer of the bladder in Quebec. Results of a multicenter survey]. PMID- 9749073 TI - [Treatment of superficial tumors of the bladder]. AB - Superficial bladder cancer includes T1, Ta, TIS transitional cell carcinomas of the TNM classification. These tumors represent a spectrum disease with different biological behavior. Some tumors may recur on the same mode for years, others have a definite metastatic risk in the short range. The main difficulty in the management of these tumors is to predict their potential aggressiveness based on clinicopathological parameters. Their management is based on the initial endoscopy and histopathological analysis. Molecular markers will reach the clinical level in order to better predict the prognosis and improved the non invasive tools for follow-up. For low risk tumors, transurethral resection (TUR) and surveillance allow an adequate tumor control. For high risk tumors, conservative treatment is based on a complete transurethral resection which need confirmation by a second look TUR and prophylactic endovesical instillations of bacille Calmette Guerin (BCG). The response to the first BCG cycle represent a crucial indicator of tumor behavior. For intermediate risk lesions, the advantage of prophylactic endovesical instillations have been finally established either using BCG or chemotherapy (Mitomycine C). However in this setting, various therapeutic modalities (maintenance therapy, dose, repeat cycles) are proposed and their relative efficacy remain to be established. For this later group of tumor it is more likely that the use of maintenance therapy or repeated courses increase the chance of conservative treatment. Other therapeutic modalities exist (oral interferon inductors, photochemotherapy) but remain second line or investigative therapies. In conclusion, progress in the understanding of the natural history of bladder cancer allow a better management of these tumor in order to optimize the ratio between the oncologic efficacy and quality of life. PMID- 9749074 TI - [Endoscopic surveillance of superficial papillary tumors of the bladder]. AB - All cases of superficial bladder tumor treated in 1991 at the urology department of La Pitie hospital were reviewed to study the course of superficial bladder tumors with regard to the risk of muscle infiltration and the value of endoscopic follow-up. In 1991, 73 patients (63 men and 10 women) with a mean age of 64 years had TUR for superficial bladder tumor. Patients had repeated cystoscopy at 3 and 6 months and then on a yearly basis for 5 years. Fourty-eight patients had TIS tumors, and 19 patients had T1 tumors. Five patients were lost to follow-up. Sixty-eight patients were followed for 5 years. Of 48 patients with stage TIS tumors, 39 (81.5%) did not show any deterioration at histological examination, eight patients (16.5%) had infiltration of the lamina propria by tumor (pT1) and the tumor invaded muscle in one patient (2%). Of the 19 patients with stage T1 tumors at the first TUR eight (47%) had no pathologic deterioration, but nine (53%) developed muscle invasion (pT2). Two patients were lost to follow-up. Invasion of the lamina propria present at diagnosis or during the subsequent course is considered to be an indicator of high risk of progression to muscle invasive disease (over 50% of cases). The prognostic accuracy of both the grade and stage needs to be enhanced using molecular markers. PMID- 9749075 TI - [Combination of endoscopic resection and external radiotherapy in carcinoma of the bladder]. AB - The authors present results that were obtained by combining endoscopic resection and external beam radiation therapy in a population of 55 patients presenting with bladder cancer. They also explain the difficulties they encountered in the management of bladder cancer at the Ibn Rochd Cancer Center (Casablanca, Morocco). PMID- 9749076 TI - [Vesical replacement after cystectomy for tumor of the bladder]. AB - Replacement of bladder after cystectomies avoids urinary diversion. It allows suboptimal natural miction, especially during the day. Current surgical techniques allow reconstruction of the bladder with detubulated ileal small bowel. This major procedure is achievable if the patient performance status is satisfactory and if urethra can be conserved. Bladder replacement is also feasible in women in selected cases. There is a need for long-term follow-up because of the risk of urological complications, especially ureteral stenosis and metabolic disorders. PMID- 9749077 TI - [Survival of patients with infiltrating tumors of the bladder treated with cystectomy]. AB - We reviewed the results of infiltrating bladder cancer treated by radical cystectomy to evaluate survival. Between January 1989 and December 1992, a total of 58 consecutive cystectomies or anterior pelvectomies performed on 48 men and 10 women (mean age 63.2 years) in our department were retrospectively evaluated. Four patients were lost to follow-up and the mean follow-up was 72 months. Pathologic staging was as follows: stage pT0,A,1: 13.5%, stage pT2: 17.5%, stage pT3a: 12%, stage pT3b: 36%, stage pT4: 21%. The year probability of the overall survival was 60% for p T2-p T3a patients, 15% for pT3b patients, and 9% for pT4 patients, respectively. Overall, 53.5% of patients died of cancer, 7.5% of intercurrent disease, and 39% were alive. The cancer related death rate was 12% for pT2-pT3a patients, and 82% for pT3b-pT4 patients. The 5- year probability of specific survival was 80% for pT2-pT3a patients, 15% for pT3b patients, and 9% for pT4 patients, respectively. Infiltrating bladder cancer still has a high mortality rate. Radical cystectomy may be considered to be a curative procedure for stages pT2 and pT3a. Adjuvant chemotherapy and/or radiotherapy seem necessary at stages pT3b and pT4. Preoperative criteria need to be better defined to reduce understaging. PMID- 9749078 TI - [Surgical treatment of cancer of the bladder]. AB - In the Saguenay-Lac-Saint-Jean area, an abnormal incidence of bladder cancer bas been identified. This incidence is an effect of the important industrial environment and the etiology has been identified. A review of the surgical treatment is actually done. Following this evaluation, we have made a choice in our medical environment (the ileal conduit or the detubularised ileal bladder for a replacement after a radical cystectomy). For the 1970 to 1997 period, a statistic statement has been done and shows the patients survival in accordance with the grade and the stage. A more appropriate treatment should be given in T3A and T3B stages. In conclusion, we have instituted a group of stimulus in the industrial environment to improve the survival for the next decades. PMID- 9749079 TI - [Role of radiotherapy in cancer of the bladder in Saguenay-Lac Saint-Jean]. AB - Bladder cancer is more frequent in Quebec, especially in Saguenay-Lac Saint-Jean than in other Canadian provinces and in the USA. From 1983 to 1996, only 78 patients presenting with bladder cancer received external beam radiation therapy. Sixty-eight were treated with curative intent. Overall survival rates were 70% at 3 years, 66% at 5 years, and 40% at 10 years. Retrospective analysis of these cases and literature review show that preoperative radiation therapy is useful in the management of bladder cancer, especially in T3 tumors. It is also useful for patients whose tumor objectively responds to radiation therapy, without an increase in morbidity. PMID- 9749080 TI - [Interstitial brachytherapy in infiltrating cancer of the bladder. The Nancy experience]. AB - PURPOSE: From 1975 to 1996, 98 patients with infiltrative vesical carcinomas were treated at the Centre Alexis Vautrin by conservative surgery and interstitial brachytherapy (192lr). The mean follow-up was about eight years. From this retrospective non randomized study, we tried to determine the tolerance to this treatment. MATERIALS AND METHODS: There were 86 men and 12 women. The mean age was 63 years. We found three pTis tumors, 28 stage pT1 tumors, 38 stage pT2 tumors, 24 stage pT3A tumors, four stage pT3B tumors and one stage Tx tumors. The therapeutic scheme consisted of pelvic radiation therapy (3 fractions of 3,5 Gy) immediately followed by lymphadenectomy (for stage pT3 tumors) and by cystotomy or partial cystectomy during which we inserted brachytherapy plastic tubes. The delivered dose was 50 Gy for superficially infiltrative tumors and 30 Gy for deeply infiltrative tumors; at the lowest dose, the treatment ended with external beam irradiation. RESULTS: At 5 years the control rate was 72%, the specific survival 80% and the global survival 71%. Twenty-nine patients had a local recurrence. Of these, seven underwent total cystectomy. Thirty-seven patients developed 43 complications; 35 were intravesical, 10 (28%) were estimated to be higher than grade 2 because of technical problems that led us to modify the technique. CONCLUSION: It is essential to develop close collaboration between surgeons and brachytherapists, to select patients and to use a rigorous technique. Interstitial brachytherapy for infiltrative vesical carcinomas thus yields both high local control and satisfying results in regard to patient's well being. PMID- 9749081 TI - [Conservative treatment of infiltrating cancer of the bladder: neoadjuvant chemotherapy and radiotherapy]. AB - PURPOSE: Retrospective evaluation of tolerance and efficacity of a combination of chemotherapy and radiotherapy in non metastatic invasive cancer of the bladder. MATERIALS AND METHODS: From 1984 to 1995, 36 patients presenting with invasive urothelial carcinoma of the bladder were treated conservatively with primary chemotherapy: cisplatin-methotrexate-vinblastine (24 patients), methotrexate vinblastine-doxorubicin-cisplatin (11 patients) or carboplatin (one patient). Fourty-six Gy were then delivered by external radiation therapy to the pelvis, with a complement of 10 to 20 Gy to the bladder. Seventeen patients received a concomitant chemotherapy with cisplatin. RESULTS: The intolerance to chemotherapy was hematologic and digestive. One patient died from bone marrow depletion. External irradiation was well tolerated, except for one patient whose treatment was stopped at 48 Gy. Fifteen tumours (44%) were in complete remission (CR) after chemotherapy and 23 (64%) at the end of treatment. With a median follow up of 64.6 months, 13 out of 23 patients in CR had a relapse (ten local relapses, three metastatic); five salvage cystectomies were done. The median survival and 26 months and the probability of survival at 5 years was 43%. All but two patients surviving 5 years had a functional bladder. CONCLUSION: Neoadjuvant chemotherapy leads to CR in 44% of patients and CR is observed in 64% of the patients after radiation therapy. However, the survival rate at 5 years is insufficient, even if the rate of bladder conservation is high. PMID- 9749082 TI - [Concurrent radio-chemotherapy in infiltrating cancer of the bladder: a new therapeutic approach?]. AB - Until now, radical cystectomy has been considered the most effective treatment for invasive bladder cancer. However, it fails to cure more than 50% of patients and can result in a mediocre quality of life. In an effort to improve cure rates, combined modality regimens have been investigated. Despite the preliminary results of early clinical trials, randomized trials have most often failed to show any benefit from neoadjuvant or adjuvant chemotherapy or radiotherapy. One of the major progress brought by radiotherapy has been the wide use of conservative treatment in several cancer, and in the recent years promising results with concomitant radiochemotherapy have been published. Use of the conservative approach in bladder cancer now appears to be a tangible reality for selected patients, but these combined modalities have not yet been tested in randomized trials. PMID- 9749083 TI - [Treatment of cancer of the bladder in elderly patients with an intra-arterial chemotherapy and radiotherapy combination: 10-year experience]. AB - PURPOSE: Analysis of the results obtained in elderly (75 years and older) included a phase II trial combining intra-arterial cisplatin and concurrent radiation into invasive bladder cancer. PATIENTS AND METHODS: Thirty-five patients (28 males and 7 females) were accrued from 1985 to 1996. There were 1 Ta, 4 T2, 11 T3A, 12 T3B, 3 T4A, and 4 T4B patients. Nine had unilateral hydronephrosis and two bilateral hydronephrosis. There were 28 transurethral resections which were incomplete in 23 patients. Intra-arterial cisplatin was given as 2-4 hours infusion (60-90 mg/m2) split through both internal iliac arteries on day 1, 14, 21, and 42. Irradiation to the pelvis was started on day 14 and consisted of 40 Gy/20 fractions followed by a boost of 20 Gy/10 fractions to the tumor with margins of 2 cm. RESULTS: Thirty (86%) completed fully the protocol. One patient died from sepsis secondary to the treatment. The tumor response was evaluable in 29 patients and complete response was observed for 27 of them. Five of these 27 patients had an isolated bladder relapse which was salvaged by cystectomy in two patients. There were 11 deaths from bladder cancer (31% of the patients): 9 from distant metastase, one from local failure, and one from treatment. CONCLUSION: This combined modality yields excellent results with high complete response rate and good tolerance. This approach may therefore be particularly appropriate for the elderly. PMID- 9749084 TI - [Treatment of infiltrating cancer of the bladder with cisplatin, fluorouracil, and concurrent radiotherapy: results of a pilot study]. AB - PURPOSE: Pilot study to assess treatment feasibility and results of a 2-drug chemotherapy (CT) regimen administered concurrently with radiotherapy (RT) for muscle-invasive bladder cancer. MATERIALS AND METHODS: Fourty-six patients were included into a prospective study from December 1992 to April 1996. The median age was 66 years. Thirty-seven percent of the patients had T3B-T4 tumors, and 46% had benefited from prior macroscopically complete transurethral resection (TUR). Pelvic irradiation consisted of 50.4 Gy in 28 fractions over 39 days. Concurrent CT consisting of cisplatin (80 mg/m2/d1) and 5-fluorouracil (800 mg/m2/d2 to 5) by continuous infusions (5-FU) was delivered during the first and fifth weeks of radiation therapy. Twenty-three patients received two additional courses of adjuvant CT with cisplatin, methotrexate and vinblastin (MCV). RESULTS: The median follow-up was 38 months. The feasibility of concurrent CT-RT was excellent: 96% of the patients completed radiotherapy and 100% of them received the two courses of P-FU. The acute toxicity was mild: no hematological toxicity or renal toxicity over grade II, 4 cases of bowel or rectal reversible grade III toxicity and 2 cases of reversible grade III cystitis. A complete response was achieved in 30 out of the 42 evaluable patients (65.2%). Nine patients received an immediate salvage treatment (3 TUR, 3 additional radiotherapy and 3 cystectomies). Ten patients had local failure. Projected 3-year locoregional control was 49% for the 46 patients. Projected overall 3-year survival was 53%. Functional results were good for disease-free patients with preserved bladder: 1 grade I, 3 grade II, and no grade III cystitis. CONCLUSION: Concurrent 2-drug chemoradiotherapy with cisplatin and 5-fluorouracil is feasible without major toxicity and offers a potentially curative and conservative treatment for patients with localized muscle-invasive bladder cancer. PMID- 9749085 TI - [Concurrent chemotherapy and hyperfractionated radiotherapy for non-operable carcinoma of the bladder]. AB - Two sequences of concomitant chemotherapy with 5-FU + cisplatin and hyperfractionated radiotherapy (67.2 Gy/56 fractions/38 days, two fractions of 1.2 Gy per day spaced to 6 hours) in the bladder volume were administered to 17 patients with infiltrative carcinoma of the bladder (1 pT1, 6 pT2, 8 pT3a, 2 pT3b or pT4, 8/17 N0-2). They were separated by an interval of 21 days with an intermediate bladder tumor evaluation. A complete histological response was observed in 69% (11 of 16 evaluable patients). With a mean follow-up of 21 months, the bladder preservation was stated in 8/11 (73%) of patients treated, of whom 6 with a complete tumor response, and 2 with a pTa recurrence; these results are similar to other series reported with conventional radiation therapy. No deficit in the bladder was observed in these eight patients. PMID- 9749086 TI - [Results of long-term treatment of inoperable cancer of the bladder with cisplatin and concurrent irradiation: prognostic factors of local control and survival]. AB - PURPOSE: Therapeutic strategies for muscle invasive bladder cancer are currently evolving. A recent European randomized study has shown that neoadjuvant chemotherapy does not improve the chance of cure or and radiotherapy would provide better results but there is a need to identify by prognostic factors patients who may benefit from such a conservative strategy. MATERIAL AND METHODS: One hundred and nine patients with localized muscle-invasive bladder cancer, who were not candidates for radical cystectomy, were treated with concomitant cisplatin and radiation therapy. Their mean age was 71. Thirty-six percent of the patients had T3B-4 tumors, and 37% had benefited from prior macroscopically complete transurethral resection (TUR). Pelvic irradiation consisted of 40 to 45 Gy and was followed by a boost to the bladder to a total dose of 55 to 60 Gy. Continuous infusion cisplatin (20 to 25 mg/m2/d for 5 days) was delivered during the second and fifth weeks of radiation therapy. RESULTS: Median follow-up was 73 months. The projected 5-year locoregional control rate was 43% for the 109 patients and 55% for the 86 patients with complete response. The projected overall 5-year survival rate was 36% for all patients and 44% for complete responders. Univariate analysis of prognostic factors was carried out for local control, and survival. The local control was statistically better in patients with good performance status, T2-3A, complete initial TUR, and in patients without hydronephrosis. In terms of overall survival, four factors were significant: the performance status, T-stage, absence of hydronephrosis, and complete response. By multivariate analysis, performance status, hydronephrosis and T-stage were significant factors for local control, while T-stage and complete response were the strongest determinants for survival. CONCLUSION: Concurrent cisplatin and radiation therapy is a potentially locally curative treatment for 43% of patients with muscle-invasive bladder cancer not candidates for radical surgery. Clinical T-stage and hydronephrosis have a significant and independent prognostic value on local control but appears not discriminant enough to select patients for conservative treatment. PMID- 9749087 TI - [Chemotherapy of metastatic cancer of the bladder. Apropos of 50 patients treated with M-VAC]. AB - The most effective chemotherapy of metastatic bladder cancer in M-VAC (methotrexate, vinblastine, cisplatin), as it was shown M-VAC by phase III trials. The complete remission rate is 15-20%. The toxicity is severe, and M-VAC cannot be delivered to fragile patients. Survival is improved. This increase in survival results in an increased incidence in brain metastases. The Cochin Hospital experience with 50 patients treated with M-VAC for a metastatic bladder cancer is presented. PMID- 9749089 TI - [Radioprotection and radiotherapy: the new regulatory texts]. PMID- 9749088 TI - [The role of new drugs in the treatment of locally advanced urothelial tumors of the bladder]. AB - Although the optimal management of muscle-invasive bladder cancer remains a continued subset of controversy, results achieved with either radical cystectomy, or radiation therapy or combinations of the two lead to cure rates of only 50% with up to 50% of patients who will develop metastases, usually within two years. For the past ten years, chemotherapy of advanced urothelial tumors has focused on cisplatin-based combinations such as methotrexate-vinblastine-adriamycin cisplatin (M-VAC), cisplatin-methotrexate-vinblastine (CMV) or cisplatin adriamycin-cyclophosphamide (CISCA). Such regimens have provided interesting results with responses rate up to 70% and some long term survival (in 10 to 20% of patients), but this has focused achieved with moderate to severe toxicity. Thus, if the chemosensitivity of advanced urothelial tumors is obvious, it is clear that a therapeutic plateau has been reached and only the search for new active agents can provide any hope to improve the results of these established regimens. Recently some single agents under investigation including taxanes (paclitaxel and docetaxel) and gemcitabine have been identified and have entered clinical development. Preliminary data of several phase II studies are now available and they appear promising, providing the basis of new combinations regimens that will attempt to improve results that have been achieved with standard regimens during the past decade. The activity of these new single agents and new combination are reviewed. PMID- 9749090 TI - Chemoradiation for malignant epithelial tumors. AB - Combinations of the chemotherapy and radiation therapy are increasingly a part of treatment strategies for cancer. Although directed at distant metastasis, they may have little effect on the primary tumor and regional lymph nodes. The local tumor must be controlled to 1) eliminate life-threatening complications, 2) decrease the risk of distant metastasis, and 3) permit organ conservation. Induction chemotherapy has been shown to be effective in decreasing distant metastasis, but it does affect local tumor control in common epithelial tumors. Concurrent single agent chemotherapy can increase local control with radiation therapy and combination chemotherapy concurrent with irradiation can both improve local control and decrease distant metastasis. Combination chemoradiation therapy has become a major part of research strategies of the Radiation Therapy Oncology Group (RTOG): it has been shown to improve local control and survival in carcinomas of the esophagus and nasopharynx. Combination chemoradiation has increased organ conservation in carcinoma of the anal canal and it is under investigation for carcinomas of the larynx (laryngeal preservation), other head and neck sites, lung, and cervix. The equation--increased local tumor control + decreased distant metastasis = increased survival--is the paradigm. Systemic therapy with cytotoxic drugs or hormones needs to address both potential sites of failure. PMID- 9749091 TI - [Radiotherapy-induced solid tumors: review of the literature and risk assessment]. AB - Although the first radiation-induced solid tumor was reported as early as 1902, the risk of second tumor has been underestimated by radiation oncologists who treated large numbers of patients with either benign or malignant diseases. Since then, numerous epidemiological studies yielded better knowledge of the risks of radiation-induced malignancies. For instance, radiation-induced tumors are the first cause of death 10 years after treatment for Hodgkin's disease. We present here a literature review of the risks of radiation-tumors in various organs, related to the irradiation dose, age, and associated diseases. The most sensitive organs are the thyroid, central nervous system, breast, bone, and lung, especially in smokers. Higher risks are observed with increasing doses, the shape of the dose-response curve depending on the tumor type, when the irradiation is performed in children or young adults and in patients with retinoblastomas. The risk of radiation-induced tumors should lead the radiation oncology committee to reconsider the dose and technique of irradiation and to reduce the use of ionizing radiation in benign diseases. PMID- 9749092 TI - [Results of the conservative surgical and irradiation treatment of 132 nonpalpable ductal carcinomas in situ of the breast]. AB - PURPOSE: Retrospective analysis of results of treatment of 132 subclinical ductal carcinomas in situ, non-palpable. MATERIAL AND METHODS: Patients were treated with limited surgery and 70 Gy radiation therapy (70 Gy). RESULTS: With a median follow-up of 7 years, the total recurrence rate was 6%, and the actuarial rate at 5 years 4% and at 10 years 13% at. These have no influence on recurrence on the specific actuarial survival rate which was 100% at 10 years. In spite of five infiltrating recurrences of seven, no metastasis appeared 48 months after the salvage surgery. The global rate of breast preservation was 92% at 7 years. DISCUSSION AND CONCLUSION: Therapeutic indications were developed taking into account the present analysis and a literature review (2,338 in situ ductal carcinomas, palpable or not, treated with conservative surgery, with or without adjuvant radio-therapy). PMID- 9749093 TI - [Clinical and biological evaluation of the response to neoadjuvant hormone therapy before radiotherapy in nonmetastatic cancers of the prostate]. AB - PURPOSE: The aim of this study was to evaluate the clinical and biochemical response to neoadjuvant hormonal therapy (NAHT) before radical external radiotherapy (EBRT). MATERIALS AND METHODS: From June 1986 to June 1994, 105 patients with histologically proven and non-metastatic prostate adenocarcinoma (stage B2-C2) received a short induction hormonal therapy (median: 3 months) with a luteinizing hormone releasing hormone (LHRH) analog associated with an anti androgen followed in all cases by EBRT (66 Gy). All patients underwent a prostate specific acid (PSA) determination, pelvic computed tomography (CT) scan and bone scan before the combined treatment. Response, treatment toxicity and PSA concentration were analyzed after the NAHT, 3 months after the completion of radiotherapy and every 6 months there after. Relapse was defined by PSA elevations above 4 ng/mL or two consecutive elevation above 1 ng/mL. RESULTS: Median follow-up was 52 months. According to the Withmore-Jewett clinical classification, 85 tumors were stage C. Pre-treatment PSA (PSAi) was above 20 ng/mL in 63.8% of the patients (median PSAi: 26 ng/mL). A clinical evaluation and a PSA determination (PSAPH) were both performed for all patients after NAHT. Most of the time, urinary obstructive symptoms disappeared with androgen ablation; tumor volume regression exceeded 50% in 99 cases and was complete in 50 cases. Median PSAPH was 0.6 ng/mL for the entire group. Clinical and biochemical tumor response were coherent: 84% of patients with clinical total remission had a PSAPH < 1 ng/mL. PSAPH value was significantly correlated with tumor stage and pre treatment PSAi: among the 11 patients with a PSAPH > 4 ng/mL, ten were stage C and nine had a PSAi > 20 ng/mL. A PSAPH value exceeding 4 ng/mL predicted biochemical relapse (P < 0.0001). CONCLUSION: We conclude that biochemical response to hormonal therapy has a major prognostic value before EBRT can help to identify patients for an adjuvant hormonal therapy. PMID- 9749094 TI - [Low-dose postoperative vaginal brachytherapy of adenocarcinoma of the endometrium]. AB - PURPOSE: Surgery is the primary treatment for endometrial carcinoma. Methods of complementary treatment are still debated, with the potential association of external radiotherapy and/or brachytherapy before or after surgery. This study was aimed at evaluating local control and complications rates in a series of patients treated by hysterectomy followed by postoperative vaginal low-dose rate brachytherapy (BT) combined with pelvic irradiation in case of poor prognosis factors. PATIENTS AND METHODS: From 1978 to 1993, 101 patients were treated at the Centre Alexis-Vautrin, France according to this scheme. Forty five had deep myometrial invasion, and thirteen cervical involvement. Fifty patients received pelvic irradiation (median dose 46 Gy) combined with BT (dose 14 Gy, median volume 127 cm3); 51 patients had BT alone (dose 60 Gy, median volume 71 cm3). RESULTS: The 5-year overall survival rate was 83% and the local control rate 97% with a median follow-up of 7 years. Multivariate analysis showed two factors of bad prognosis, i.e., deep myometrial invasion and cervical involvement. Three severe complications occurred in two patients for whom the treated volume was larger than the theoretical target volume. Eleven patients developed metastases. CONCLUSION: Results obtained from this series are comparable with those of previous studies, particularly in regard to pre-operative BT. The complication rate is also satisfactory and depends on the irradiation precision and the definition of the target volume. PMID- 9749095 TI - [Is cerebral tomoscintigraphy with 99mTc-MIBI useful in the diagnosis of local recurrence in patients with malignant gliomas?]. AB - PURPOSE: 99mTc-MIBI, an alternative radiopharmaceutical for myocardial perfusion study has been proposed for use as a tumor imaging agent, including breast cancer, lung cancer, lymphomas, melanomas, and brain tumors. After routine radiation therapy, deteriorating clinical status or treatment failure may be due to either radiation changes or recurrent tumor. CT and MRI offer imperfect discrimination of tumor viability and radionecrosis. MATERIALS AND METHODS: Thirty-five malignant glioma patients with clinical deterioration were studied retrospectively. Tomoscintigraphy was performed 15 minutes after intravenous injection of 1110 Mbq 99mTc-MIBI. The images were obtained from a dual headed gamma camera using fan beam collimator. Transverse, coronal and sagittal views were reconstructed. RESULTS: A dramatic MIBI uptake was found in 31 patients. This uptake was correlated to tumor recurrence proven by histological fragments and/or the rapid, fatal evolution of these patients. Death occurred after the brain SPECT had been performed for those cases showing MIBI uptake, an average 5.48 months later. No MIBI uptake was found for these four remaining patients: their evolution can be currently considered to be a disease-free time. CONCLUSIONS: According to our results, the sensibility and specificity of 99mTcMIBI brain SPECT seems to be high. Moreover, this investigation is more accurate for discriminating tumor recurrence from radionecrosis than a CT scan or MRI. PMID- 9749096 TI - [Radiotherapy of epidemic Kaposi's sarcoma: the experience of the Henri-Mondor Hospital (643 patients)]. AB - PURPOSE: Retrospective analysis of results of radiotherapy in epidemic Kaposi sarcoma at the Henri-Mondor hospital. MATERIAL AND METHODS: From June 1986 to December 1996, 643 patients presenting with acquired immunodeficiency syndrome (AIDS)-related epidemic Kaposi's sarcoma were treated with irradiation at the Oncology Department of Henri Mondor University Hospital. Three-hundred eighty seven patients (60%) had previously received a treatment with interferon (259 patients, 40.2%), vinblastine (225 patients, 34.5%), doxorubicin (22 patients, 3.4%), bleomycin (19 patients, 2.9%), and/or antiviral treatment (216, 33.5%). The radiotherapy was delivered by 4 MeV or 8 MeV electron beam for extended cutaneous fields and 45-100 kV x-ray for localized fields. The delivered dose was 20 Gy in 2 weeks (2.5 Gy/fraction, 4 fractions/week) followed by 2 weeks rest and second series of 10 Gy in 1 week. For oral cavity lesions, we used a series of 15.2 Gy was delivered (1.9 Gy/fraction, 4 fractions/week), followed for three patients by a 3 week rest and by a similar second series of 15.2 Gy. RESULTS: Six hundred and twenty-one patients were evaluable and the objective response rate was 92%, with a complete regression of clinical and functional symptoms for all patients. The skin tolerance was good, with 7.3% grade I reactions, 69.3% of grade II reactions, and 23.4% grade III reactions. There was a correlation between recurrence rate and the occurrence of opportunistic infections. CONCLUSION: This analysis shows the efficacy of dose radiotherapy for treatment of epidemic Kaposi sarcoma. PMID- 9749097 TI - [Evaluation tests of computer systems concerning tri-dimensional dose calculations]. AB - The development of irradiation techniques in radiotherapy shows a clear tendency towards the systematic use of three-dimensional (3D) information. Great efforts are being made to set up 3D conformal radiotherapy. Consequently, in the aim of greater coherence and accuracy, "the dosimetric tool" must also meet the requirements of 3D radiotherapy, as it plays a role in the treatment chain. To know if the treatment planning system is a "3D", "2D" or even "1D" system, one should not be satisfied with reading the technical documentation and the program algorithm description nor entirely trust the constructor's assertions. It is essential to clearly and precisely evaluate the possibilities of the treatment planning system. Even if it is proved not to satisfy perfectly all the tests which would qualify it as a real 3D calculation system, the study of the test results helps to give clear explanations of the dosimetric results. Two series of test cases are proposed. The first series allows us to understand in which conditions the treatment planning system takes into account the scatter influence in a volume. The second series is designed to inform us about the capability of the dose calculation algorithm when the medium encloses non-homogeneities. These test cases do not constitute an exhaustive "check-list" able to tackle completely the question of 3D calculation. They are submitted as examples and should be considered as an evaluation methodology for the software implanted in the treatment planning system. PMID- 9749098 TI - [39th Meeting of the American Society for Therapeutic Radiology and Oncology, Orlando (Florida), 19-23 October, 1997]. PMID- 9749100 TI - [Conformal radiotherapy in cancer of the prostate]. PMID- 9749099 TI - [9th European Cancer Conference: EECO 9 Hamburg (Germany), 14-18 September, 1997]. PMID- 9749101 TI - [General principles of conformal radiotherapy]. PMID- 9749102 TI - [Indications for tridimensional conformal radiotherapy in cancer of the prostate]. PMID- 9749103 TI - [Conformal radiotherapy: arguments against]. PMID- 9749104 TI - [History of radiosurgery]. PMID- 9749105 TI - [Dosimetry of small-size photon beams]. AB - A high performance standard radiotherapy treatment unit could be used to perform stereotactic radiosurgery. The dosimetric aspects of stereotactic irradiation with small size photon beams (energies from 5 to 25 MV produced by electron linear accelerator or gamma-rays produced by cobalt-60 treatment unit) are analyzed. The diameter of circular beams used are 5 to 40 millimeters wide at the isocenter of the treatment unit. The dosimetry of small x-ray fields is complicated by two factors: the relationship between detector size and field size dimensions, and the lack of lateral electron equilibrium. The large dose gradients outside the beam's central axis require dosimetry techniques with higher spatial resolution. To determine the best dosimetry system for measurements at the beam's small focal point, particularly for measurement of the field size dependent on output factors, several different detectors were investigated: ionization chamber, silicon diode, diamond detector, thermoluminescent dosimeter, and film. Ionization chamber, which presents a sensitive volume smaller than 0.02 cm3, is the most commonly recommended detector for field diameter above 8 mm. Current representative examples of dosimetric measurements for different x-ray energies, including percent depth dose, tissue maximum ratios, beam profiles (off axis ratios), and output factors, are presented and discussed. As well, the dosimetric characteristics of small photon beams are detailed. PMID- 9749106 TI - [Progress in dosage optimization for stereotactic radiosurgery]. AB - Stereotactic radiosurgery is a technique for treatment of intracranial lesions requiring high precision in all steps--from image acquisition to final irradiation. One of most difficult steps is the treatment planning phase, consisting of determination of irradiation parameters sufficient to cover the target volume by avoiding sensitive volumes. A manual and empirical definition can be very long and difficult, especially in the case of complex target volumes situated in sensitive zones. As in conventional radiotherapy, stereotactic radiosurgery has taken advantages from dosimetric optimization. The question is: "What is the configuration of irradiation parameters used in order to obtain the treatment plan by satisfying defined constraints?". The purpose of this article is to summarize optimization methods used in radiosurgery and to describe the technical alternatives proposed for this treatment as well as the possibilities of plan evaluation between different techniques. This purpose will be illustrated by the optimization methodology used in the Center Oscar Lambret of Lille, France for the radiosurgical treatment with linear accelerator. PMID- 9749107 TI - [Progress in optimizing dosimetry plans in stereotactic radiotherapy in the Salt Group (Saint-Anne-Lariboisiere-Tenon]. AB - We began intracranial stereotactic irradiation under the direction of O Betti 11 years ago. At the present time, we believe it is interesting to present the methodologies of the SALT (Saint-Anne-Lariboisiere-Tenon) group. Up to the present time we have irradiated 693 patients using a single fraction. Arteriovenous malformations (AVMs) represented the majority (90%) of treated lesions. Irradiation protocol has little changed since 1986, and the localization of the target volume was performed in the neurosurgery department of St Anne Hospital, France. The stereotactic images (computerized tomography [CT], angiography) were sent to the radiotherapy department of Tenon Hospital through the French public digital network NUMERIS. Protocol was realized using the stereotactic ARTEMIS-3D/Dosigray TPS. The lesion volume was filled by one or more spherical or elliptical subvolumes using the "Associated Target Methodology". The interactive adjustment of subvolumes was based on the 3D graphical representations of the lesion. The direct optimization of the irradiation space was performed by managing parameters provided by the DDB (Dosimetric Data Base) such as the number of arcs, their angular position, as well as the starting and the ending point of each arc. The evaluation of the calculated dose distribution was made using quantitative parameters. The second method of optimization was based on the minibeam intensity modulation using a mathematical theory of inverse problems and singular value decomposition (SVD) analysis. At the present time, due to technical reasons, linear accelerators do not permit the modulation of intensity of arctherapy. Thus we transformed the profiles of irregular forms into rectangular profiles of modulated ponderation, with each optimized plan being evaluated before its implementation. The criteria of evaluation were derived from the differential and cumulative dose volume histograms (DVH). The DVHs permitted the evaluation of the volumes of underdosage and overdosage inside the lesion and in the healthy tissue, respectively. Using DVHs, we have defined parameters such as the conform factor and the homogeneity index. We stress that the methodology of protocol optimization is valid for single or multiple fractions as well as for intra- and extra-cranial irradiation. PMID- 9749108 TI - [Dosimetric quality control in stereotactic radiotherapy with the aid of radiosensitivity]. AB - Typical dosimeters used in stereotactic radiation therapy, such as ionization chambers, films, and thermoluminescent diodes, allow basic physical measurements. They are, however, neither well suited to discern small target volumes with high dose gradient, nor suitable for three-dimensional (3D) dose measurements. Gel dosimetry is becoming more and more interesting, owing to magnetic resonance imaging (MRI). It permits isocenter position planning verification of accuracy and the precision of the 3D dose mapping in the brain (when irradiated in realistic conditions), especially when several different targets are concerned. Many authors have assessed stereotactic radiation therapy quality control using different gels, and different irradiation procedures. This paper consists of the review of these different methods to assess quality control. Gel dosimetry cannot provide absolute dose measurements. However, gels can be used to check the 3D dose mapping with a high degree of detail. In our experiment, the difference between the stereotactic frame center and the isocenter is about 1 mm. The difference between the theoretical isodoses obtained by the treatment planning system and the experimental isodoses obtained by the MRI gray level calibration is also about 1 mm, the order of magnitude of the MRI pixel size. PMID- 9749109 TI - [Stereotactic localization in medical imaging. Technical and methodologic aspects]. AB - Stereotactic neurosurgery and stereotactic radiation therapy require the three dimensional localization of lesions for biopsy or for treatment planning. The aim of this paper is the description of methods used in the different imaging modalities: x-ray teleradiography, digital subtracted angiography, computed tomography, and nuclear magnetic resonance imaging. The simple pin-target locating techniques are distinguished from those serving to the definition of volumes target necessary to treatment planning. Performances and difficulties of these techniques are emphasized. The specific methodology developed in Lille is described as an example. Organizational aspects and necessary quality controls for a good progress of the entire procedure, from imaging to treatment, are also discussed. PMID- 9749110 TI - [Technical evolution of irradiation in stereotactic conditions: dose fractionation]. AB - The development of non-invasive head fixation systems, allowing 3D determination of the target coordinates, has lead to the increased use of fractionated stereotactic irradiation. These systems have been checked for accuracy and the mean precision of repositioning has been evaluated to +/- 1 mm. With the mean geometrical accuracy set at +/- 1 mm, a 2 mm safety margin is usually added to the clinical target volume in order to define the planning target volume. Quality assurance procedures must conform to the required precision of the technique while remaining realistic in day-to-day use relative to planned conventional treatments. Biologically different from single dose irradiation, the fractionated stereotactic irradiation completes the range of techniques used in the treatment of intra-cerebral lesions. PMID- 9749111 TI - [Radiosurgical activity in France. Surveys of the French Society of Oncologic Radiotherapy (SFRO)]. AB - The total record of 3,407 radiosurgical treatments performed from 1986 to 1996 is described per center and per diagnosis. Treatment of arteriovenous malformation (AVM) remains the principal indication (approximately 40%) before treatment of metastasis (approximately 20%). Annual incidence of radiosurgery for AVM can be estimated to 5 per million inhabitants per year in France. First therapeutic results of AVM series are described. PMID- 9749112 TI - [Radioanatomy of cerebral arteriovenous malformations]. AB - New imaging modalities permit detailed knowledge of the anatomy of cerebral arteriovenous malformations. Magnetic resonance imaging (MRI) provides morphological data, size and topography of the nidus, anatomic relationship, as well as dynamic information particularly with the use of MR angiography. Selective and hyperselective cerebral angiography provide information about the angioarchitecture and search for associated vascular abnormalities such as aneurysms. It is therefore possible to distinguish malformations associated with a high risk of hemorrhage and to define the indications for radiosurgery. PMID- 9749113 TI - [Radiosurgery of cerebral arteriovenous malformations. Therapeutic strategies]. AB - Modalities of management of arterio-venous malformations are discussed according to natural history and size of lesion, patient clinical status, and potential side effects of available techniques (microsurgery, radiosurgery, ambolization). Clinical cases illustrate this discussion. PMID- 9749114 TI - [Radiosurgery or microsurgery of vestibular schwannomas?]. AB - PURPOSE: Microsurgical excision is the reference treatment of vestibular schwannomas. Yet, morbidity and functional risk of this surgery is significant, as Pellet et al have demonstrated in their study. In order to define the role of gamma knife surgery we have designed a prospective study concerning effectiveness and tolerance of this treatment. PATIENTS AND METHODS: Between July 14, 1992 and August, 1997, 400 patients were included. All the treatments were carried out according to a homogeneous methodology, with multiisocentric planning. We use low marginal doses dependent mainly on the treatment volume: 14 Gy for small stade II tumors, 12 Gy or less for larger tumors and 16 Gy for intracanalicular tumors. The evaluation protocol included preoperative examination with clinical examination, House grading, Shirmer's test, tonal and vocal audiometry, brain stem electrical response audiometry (BERA), vestibular caloric and pendular tests, magnetic resonance imaging (MRI) or computerized tomography (CT) scan; control at 6 months, 1 year and 2 years with clinical examination, tonal audiometry and MRI and/or CT scan; at 3 years the preoperative examination was repeated and a questionnaire based on Pellet's concerning functional results was completed. RESULTS: Among the 400 treatments, 80% were first intended treatment of unilateral vestibular schwannoma. At the time of the analysis, 100 consecutive patients with unilateral schwannoma (treated in first intention) had a 3 year follow-up (results concerned these 100 patients). Average age was 61 years (17-82 years). According to Portmann's classification, five patients presented a stade 1 tumor, 60 a stade 2 tumor, 33 a stade 3, and two a stade 4 tumor. According to House's grading at preoperative examination, there was 86 (86%) grade 1 tumors, 12 grade 2, two grade 3, and at 3 year follow-up: 77 (94%) grade 1 and five grade 2 (17 patients had no House grading). At preoperative examination, three patients presented an hemispasm; at 3 years this had disappeared for all patients. Two others patients presented a transient hemispasm at 8 and 11 months, respectively. At preoperative examination, four patients presented with facial numbness and 14 with hypoesthesia. At 3 year follow-up, trigeminal function was normal for all patients but one, for whom this had only improved. Seven others patients presented a transient numbness or hypoesthesia. At preoperative examination, five patients presented hydrocephalus without cerebro spinal fluid (CSF) shunting. At 3 year follow-up, seven patients presented CSF shunting: three presented a preoperative hydrocephalus, three a hydrocephalus after treatment and one a hydrocephalus secondary to tumor growth. Average age for these six patients was 71 years. Audiological classification was based on Gardner-Robertson's. Seventy percent of patients with normal hearing maintained useful hearing, and 50% of patients with useful hearing maintained serviceable hearing. Three (3%) patients (two with stade 2 tumors and one with a stade 3) had microsurgical excision at 16, 35 and 36 months, respectively. During microsurgical excision no unusual difficulty was encountered. Seventeen questionnaires investigating functional outcome and quality of life were completed: 100% (63% in Pellet's study) of the patients answered they had no facial motion disturbance, 49% (17% in Pellet's study) had no ocular problems, 20% (in Pellet's study 55%) had subjective trigeminal problems, 8% (in Pellet's study 13%) had deglutition problems, none (16% in Pellet's study) had other eating problems, 91% (61% in Pellet's study) had no change in their life. Mean hospitalization stay was 3 days (for 23 days in Pellet's study). All the patients who worked, except one, had the same professional activity (66% in Pellet's study). Mean work cessation was 7 days (130 days in Pellet's study). (ABSTRACT TRUNCATED) PMID- 9749115 TI - [Radiosurgery of brain metastasis]. AB - Radiosurgery has recently provided an alternative to conventional therapy for the treatment of brain metastases. This non-invasive technique delivers a single large fraction of ionizing radiation to a well-defined small intracranial target as brain metastases. After a computerized tomography (CT) with stereotactic frame in place for tumor localization, a dosimetric study was performed. The sharp dose gradient of radiation reduces the dose to the surrounding normal structures at a minimal level (> 10%). The prescribed dose at the periphery of the lesion varies from 8 to 27 Gy with a combined whole brain irradiation and from 20 to 35 Gy without any irradiation. Radiosurgery has been reported to be highly efficacious with a local control rate of 86% (not increased size without local recurrence). Brain metastases from melanoma and renal carcinoma are usually resistant to conventional irradiation and are highly sensitive with this technique. The morbidity is very low with a symptomatic edema rate of 5-10% at 2 years, resolved with corticosteroids. A radiation necrosis has been reported in less than 5% of cases. The patients with a good performance status, without any extracranial metastasis and with a solitary brain metastases have presented the best survival rate. New brain metastases have occurred in 20 to 30% of the cases during the follow-up. Eleven to 25% of patients died from their intracranial disease and the others from the extracranial evolution of the cancer. The median survival was still poor, ranging from 8 to 12 months. Radiosurgery is a good choice for surgically inaccessible and recurrent tumors. It represents an alternative to the neurosurgery with or without whole brain irradiation, taking into account different prognostic factors and morbidity rate. The local control and the survival rates without neurologic symptom should be considered the major endpoints of different ongoing randomized studies for evaluating the role of the radiosurgery. PMID- 9749116 TI - [Fractionated stereotactic radiotherapy: results in hypophyseal adenomas, acoustic neurinomas, and meningiomas of the cavernous sinus]. AB - PURPOSE: In order to optimize cerebral benign tumor irradiation, fractionated stereotactic radiotherapy allows a focused-volume irradiation (2.1 cm3, 16 mm diameter) under standard fractionation conditions. Results of a retrospective and multicentric analysis are presented. PATIENTS AND METHODS: Fractionated stereotactic radiotherapy uses the ballistic principles of the radiosurgery: stereotactic localization, multi-beam irradiation, secondary collimation, three dimensional dosimetry. Standard fractionation is possible with a re-locatable non invasive stereotactic device. The technique has been used for treating pituitary adenomas (86 patients), acoustic neuromas (32 patients) and cavernous meningiomas (26 patients). RESULTS: 1) pituitary adenomas: cumulative tumoral objective response rates (42 patients) were respectively 42%, 69% and 88% at 24, 48 and 60 months. The cumulative endocrinologic objective-response rates (32 patients) were respectively 53%, 75% and 85% at 24, 48 and 60 months. The cumulative risk of radio-induced hormonal deficiency varied from 18% (growth hormone [GH]) to 42% for TSH (thyroid stimulating hormone) at 48 months. No other complication was observed; 2) acoustic neuromas: 33 tumors, < 25 mm in diameter, were treated in 32 patients. Tumor control was observed in 29/33 tumors: 14 were stable, 15 decreased and three progressed. Useful hearing was maintained in 9/10 patients. Only three patients (9%) presented persistent complications; 3) cavernous meningioma: 17/19 clinical responses were noted, 20 tumoral stabilisations, one partial response and one progression (22 magnetic resonance imaging [MRI] evaluable patients). One unilateral radio-induced blindness was observed. CONCLUSION: For these benign tumors, the focused target volume obtained by the fractionated stereotactic radiotherapy seems to be better adapted to the treatment of limited benign tumors than standard radiotherapy. The use of standard fractionation reduces the risk of severe normal tissue damage, sometimes observed for radiosurgery and inherent in the use of single fraction. PMID- 9749117 TI - [Radiotherapy in stereotactic conditions (radiosurgery) in malignant brain tumors: clinical research]. AB - We have looked for trials which are in progress in the field of stereotactic radiotherapy (radiosurgery) of malignant brain tumors. Most randomized trials are conducted by the Radiation Therapy Oncology Group (RTOG) or the European Organization for Research and Treatment of Cancer (EORTC) and assess the role of radiosurgery in treatment of high grade glioma and brain metastasis. PMID- 9749118 TI - [Point of view of the radioneurologist]. AB - Since the 1970s, neuroradiology has benefited from significant advances and has become less and less invasive. SALT group (Saint-Anne-Lariboisiere-Tenon), created in 1986, treats and follows patients presenting with arteriovenous malformation with surgery, embolization and/or radiosurgery. Treatment failures and complications are analyzed in order to better define indications and improve techniques of treatment which benefit from advances in equipment and software, as well as in radiobiology and genetics. PMID- 9749120 TI - [The point of view of the radiotherapist]. AB - During the last decade, stereotactic radiotherapy has widely improved in France. Thus one should study the present situation and its future trend. QUANTITATIVE NEED: Considering single dose radiotherapy, there are about 900 to 1,000 cases treated per year. However, the trend towards more fractionated treatment will disturb this temporary equilibrium; thus more machine time will be necessary. QUALITATIVE NEED: Stereotactic radiotherapy is practiced by multi-disciplinary teams including physicians, physicists and scientific specialists. Radiotherapists and physicist are responsible for treatment planning and evaluation as well as for clinical and methodological research. Accordingly, they should possess computers, treatment planning systems, etc. Such teams are necessary to carry out complex irradiations. GENERAL EVOLUTION: Fractionation of irradiation nowadays seems mandatory for most intracranial tumors except metastases and small regular arteriovenous malformations. Heterogeneity of lesion dose is related to the geometry and the physics of convergent fixed or mobile beams. It can be improved and the healthy tissue irradiation can be diminished using the multi-isocentric planning for complex lesion or with micro multi leaf collimators. MODALITIES OF STEREOTACTIC RADIOTHERAPY ACCORDING TO LESION TYPE: For neurinomas of the acoustic nerve, fractionated stereotactic radiotherapy yields few of the complications published after single dose stereotactic radiotherapy. The same can be said for meningiomas although some series reported very few complications after single dose stereotactic radiotherapy. Solitary metastases without systemic evolution, not situated on the mid-line, are favorable candidates for palliative single dose stereotactic radiotherapy. The conjunction with total brain irradiation seems to be useful. Small arteriovenous malformations will be treated with single dose stereotactic radiotherapy, whereas voluminous and/or geometrically complex nidus could benefit from protons or photon beams modulated by micro multi leaf collimators and a few fractions. EXTRA CRANIAL STEREOTACTIC RADIOTHERAPY: Single dose stereotactic radiotherapy and fractionated stereotactic radiotherapy will be used as boost in various situations such as massif facial and in all sorts of tumors in the body specially when lesions are close to critical organs. PMID- 9749119 TI - [Point of view of the neurosurgeon]. AB - Stereotactic radiosurgery, a term introduced by Leksell, was born more than 40 years ago, but has made great strides for the last 15 years. There is no consensus among neurosurgeons as to the best device (gamma knife, linear accelerator), the treatment doses, and the clinical indications of stereotactic radiosurgery. Therefore, this report is the viewpoint of one neurosurgical team only. In the radiosurgery literature, there is no clear evidence of better results with the gamma-knife or the linear accelerators. With regard to clinical applications, cerebral arteriovenous malformations are the more widely accepted indications of radiosurgery, since a cerebral angiography can confirm the disappearance of the nidus of an arteriovenous malformation treated by stereotactic radiosurgery. Usually, small and deep arteriovenous malformations can be treated by stereotactic radiosurgery only. Nevertheless, the treatment of the other arteriovenous malformations more often require procedures including one or several of the following treatment methods: microneurosurgery, interventional neuradiology, stereotactic radiosurgery. Stereotactic radiosurgery in acoustic schwannomas, skull base meningiomas, especially those involving the cavernous sinus, brain metastases, and pituitary tumors seem attractive. Contrary to arteriovenous malformations, the lack of criteria for cure requires a long follow up and carefully controlled trials to confirm the efficiency of stereotactic radiosurgery for these tumors. On the other hand, experience of stereotactic radiosurgery for astrocytomas and glioblastomas is limited, and few publications are available. Furthermore, because of the infiltrating growth, a major impact of stereotactic radiosurgery for these tumors is presumably not to be expected. Lastly, a close multidisciplinary approach seems absolutely necessary to define the best indications of stereotactic radiosurgery and to improve its clinical results. PMID- 9749121 TI - [Stereotactic radiotherapy: nomenclature of acts]. AB - Acts of neurosurgery, neuroradiology and radiation therapy are not always identified in the French "nomenclature generale des actes professionnels" (NGAP) (general nomenclature of professional acts). Project of a new "nomenclature commune des actes professionnels" (NCAP) (communal nomenclature of professional acts) is described and discussed. PMID- 9749123 TI - [Limitations and perspectives of postoperative radiotherapy in bronchial cancer]. AB - The role of postoperative irradiation for lung cancer remains a controversial issue. The available data suggest a reduction in local relapse in cases of positive mediastinal lymph node, but how this benefit translates into survival is not known. The current indications include tumors with positive mediastinal lymph node and incomplete resection with micro- or macroscopical residue. Nevertheless, postoperative irradiation requires a meticulous technique to avoid inducing life threatening complications to vital organs such as the heart or the lung. PMID- 9749122 TI - [Total body irradiation: current indications]. AB - The choice of dose and fractionation for total body irradiation is made difficult by the large number of considerations to be taken into account. The outcome of bone marrow transplantation after total body irradiation can be understood in terms of tumour cell killing, engraftment, and normal tissue damage, each of these endpoints being influenced by irradiation-, disease-, transplant-, and patient-related factors. Interpretation of clinical data is further hampered by the overwhelming influence of logistic constraints, the small numbers of randomised studies, and the concomitant variations in total dose and fraction size or dose rate. So far, three cautious conclusions can be drawn in order to tentatively adapt the total body irradiation schedule to clinically-relevant situations. Firstly, the organs at risk for normal tissue damage (lung, liver, lens, kidney) are protected by delivering small doses per fraction at low dose rate. This suggests that, when toxicity is at stake (e.g., in children), fractionated irradiation should be preferred, provided that interfraction intervals are long enough. Secondly, fractionated irradiation should be avoided in case of T-cell depleted transplant, given the high risk of graft rejection in this setting. An alternative would be to increase total (or fractional) dose of fractionated total body irradiation, but this approach is likely to induce more normal tissue toxicity. Thirdly, clinical data have shown higher relapse rates in chronic myeloid leukaemia after fractionated or low dose rate total body irradiation, suggesting that fractionated irradiation should not be recommended, unless total (or fractional) dose is increased. Total body irradiation-containing regimens, primarily cyclophosphamide/total body irradiation, are either equivalent to or better than the chemotherapy-only regimens, primarily busulfan/cyclophosphamide. Busulfan/cyclophosphamide certainly represents a reasonable alternative, especially in patients who may not be eligible for total body irradiation because of prior irradiation to critical organs. PMID- 9749124 TI - [Preoperative concurrent radiochemotherapy for cancer of the rectum]. AB - PURPOSE: To evaluate retrospectively treatment-related morbidity of concurrent radiotherapy and chemotherapy for rectal cancer. PATIENTS AND METHODS: Between 1992 and 1995, 38 patients (median age: 60) were treated for locally advanced resectable rectal cancer. Median dose of radiotherapy was 45 Gy/25 fractions/5 weeks. Chemotherapy consisted of two courses of 5-fluorouracil and leucovorin administered during the first and the fifth weeks of radiotherapy. Median dose of 5-fluorouracil was 350 mg/m2/day, and median dose of leucovorin was 20 mg/m2/day, day 1 to day 5. Surgery was performed 5 weeks after completion of radiotherapy. RESULTS: Before surgery, one patient died of febrile neutropenia and sepsis after two cycles of chemotherapy and 45 Gy. Main pre-operative grade 3-4 toxicities were respectively: neutropenia: 3%; nausea/vomiting: 3%; diarrhea: 3%; proctitis: 5%; radiation dermatitis: 8%. Twenty-six patients underwent a low anterior resection and 11 an abdomino-perineal resection. A temporary colostomy was performed in 12 patients. Pathologic complete response rate was 27%. There was one post-operative death due to thromboembolic disease. Major post-operative grade 3-4 complications were: pelvic infection: 14%; abdominal infection: 5%; perineal sepsis: 8%; anastomotic dehiscence: 8%; cardiac failure: 5%. Delayed perineal wound healing was observed in six patients. No significant prognosic factor of post-operative complications has been observed. Median duration of hospitalization was 22 days. With a median follow-up of 24 months, 2-year overall and disease-free survival rates were 82 and 64%. CONCLUSION: Tolerance of preoperative concurrent chemoradiotherapy was acceptable. Ongoing controlled studies will assess the impact of this combined treatment on survival. PMID- 9749126 TI - [Importance of radiotherapy in stereotactic conditions (radiosurgery) in brain metastasis: experience and results of the Hopital Pitie-Salpetriere Group]. AB - PURPOSE: Retrospective analysis of the influence of clinical and technical factors on local control and survival after radiosurgery for brain metastasis. PATIENTS AND METHODS: From January 1994 to December 1996, 42 patients presenting with 71 metastases underwent radiosurgery for brain metastasis. The median age was 56 years and the median Karnofsky index 80. Primary sites included: lung (20 patients), kidney (seven), breast (five), colon (two), melanoma (three), osteosarcoma (one) and it was unknown for three patients. Seventeen patients had extracranial metastasis. Twenty-four patients were treated at recurrence which occurred after whole brain irradiation (12 patients), surgical excision (four) or after both treatments (eight). Thirty-six sessions of radiosurgery have been realized for one metastasis and 13 for two, three or four lesions. The median metastasis diameter was 21 mm and the median volume 1.7 cm3. The median peripheral dose to the lesion was 14 Gy, and the median dose at the isocenter 20 Gy. RESULTS: Sixty-five metastases were evaluable for response analysis. The overall local control rate was 82% and the 1-year actuarial rate was 72%. In univariate analysis, theoretical radioresistance (P = 0.001), diameter less than 3 cm (P = 0.039) and initial treatment with radiosurgery (P = 0.041) were significantly associated with increased local control. Only the first two factors remained significant in multivariate analysis. No prognostic factor of overall survival was identified. The median survival was 12 months. Six patients had a symptomatic oedema (RTOG grade 2), only one of which requiring a surgical excision. CONCLUSION: In conclusion, 14 Gy delivered at the periphery of metastasis seems to be a sufficient dose to control most brain metastases, with a minimal toxicity. Better results were obtained for lesions initially treated with radiosurgery, theoretically radioresistant and with a diameter less than 3 cm. PMID- 9749127 TI - [Treatment of A2-B2 prostatic cancer: results of a hormonoradiotherapy combination in the PSA era]. AB - PURPOSE: Prospective analysis of results of combined neoadjuvant hormonetherapy and external beam radiation therapy in A2-B2 prostate adenocarcinoma. PATIENTS AND METHODS: Between 1986 and 1994, 36 patients with clinical stage A2 (five patients), B1 (12 patients) and B2 (19 patients) N0 adenocarcinoma of the prostate declined for radical surgery, underwent a brief neoadjuvant hormonal therapy before external beam radiotherapy at our radiation therapy department. They all had a PSA determination before the combined treatment and no evidence of local extension or metastatic spread. They were followed clinically and with serial PSA levels for a median time of 58 months. Relapse was defined by a PSA level > or = 2.5 ng/mL. RESULTS: Median pre-treatment PSA level was 16.5 ng/mL; 16 patients had less than 15 ng/mL. Combined treatment was very well tolerated. After 3 months of neoadjuvant hormonetherapy, digital rectal examination was normalized in 27 cases with a PSA value < or = 1 ng/mL in 23. Only four tumors have relapsed (one local failure, two metastases and one PSA failure). The single factor that predicted biochemical relapse was pre-treatment PSA level: the 5-year actuarial rate of PSA failure when PSA level < 15 ng/mL was 0% and 27.5% if it was > or = 15 ng/mL (p = 0.05). During follow-up, only two patients suffered grade 2 rectitis and seven complained a total impotency. CONCLUSION: This limited study advocates hormonal neoadjuvant therapy and radiotherapy association in intracapsular prostatic carcinoma in patients declined for surgery or when pre treatment PSA is above 15 ng/mL, with mild acute and late toxicity. PMID- 9749128 TI - [Treatment of inoperable non-small cell lung cancer: induction chemotherapy or hyperfractionated radiotherapy? Apropos of 2 recent therapeutic trials]. PMID- 9749125 TI - [Treatment of cancers limited to the uterine cervix with simple hysterectomy using a vaginal approach after brachytherapy]. AB - PURPOSE: In patients with early cervix carcinoma, both radiotherapy and surgery or combined modalities provide effective therapies. In the two last modalities, recommended surgery is radical hysterectomy. The purpose of this prospective study was to assess the value of a limited vaginal hysterectomy after brachytherapy in patients without any unfavorable prognostic factor. PATIENTS AND METHODS: Twenty-two patients (stage Ia2 with vascular invasion: three patients, stage Ib 1:19 patients) with 1 cm median maximal tumor size and with previous negative laparoscopic lymphadenectomy (median number of lymph nodes: 12) underwent a limited vaginal hysterectomy 6 weeks after utero-vaginal brachytherapy. RESULTS: Two mild intra-operative complications were noted. Venous hemorrhage (100 mL) occurred in one patient during lymphadenectomy and another patient presented bladder injury during hysterectomy. These two complications were successfully controlled with no need for laparotomy. Only one late complication was observed: bladder grade G2 (b). With a 29 months follow-up (20 48 months), no recurrence was reported. CONCLUSION: These results appear promising in patients with very early cervix carcinoma but remain to be confirmed on a larger scale. PMID- 9749129 TI - [3rd Colloquium of Fundamental and Applied Radiobiology: "from radiobiology to the clinic". Sophia-Antipolis, 15-20 June, 1997]. PMID- 9749130 TI - [Brachytherapy in France in 1995. Final results of the national survey]. AB - A survey questionnaire was sent to the 189 French departments of radiation Oncology and 166 responded by brachytherapy and 358 shielded rooms were available. In Low Dose Rate (LDR) 81 departments used Cesium sources (159 afterloaders, 1,060 sources). Iridium wires were used by 84 departments (673 meters used). Only six departments used other elements. Twenty-six departments were equipped with high dose rate after loaders (HDR) all of them also using LDR techniques for most of the patients. A total of 9,160 patients were treated: 7,868 with LDR and 1,292 with HDR. The common sites treated by LDR were uterovaginal (4,300), breast (1,415), head and neck (1,049), skin (610), anorectal (220) and urologic (70). HDR was used for vaginal cuff (628), bronchi (371), oesophagus (232). PDR just started (33 patients) for a feasibility trial. The rate of patients treated by brachytherapy is around 6-8% of the irradiated patients, but the indications vary is each department. The diffusion of the techniques, and new indications should increase the number of patients being treated by brachytherapy. PMID- 9749131 TI - [Report of French radiotherapy scientific publications. Years 1995 and 1996]. PMID- 9749132 TI - [Improved survival in patients with locally advanced prostate cancer treated with radiotherapy and goserelin]. PMID- 9749133 TI - [Prophylactic cranial irradiation is indicated following complete response to induction therapy in small cell lung cancer: results of a multicentre randomized trial]. PMID- 9749135 TI - On some clinically useful measures of the accuracy of diagnostic tests. PMID- 9749134 TI - [Prevention of heterotopic ossification about the hip: final results of two randomized trials in 410 patients using either preoperative or postoperative radiation therapy]. PMID- 9749137 TI - [Can echocardiography identify mildly hypertensive patients at high risk, left untreated, based on current guidelines?]. AB - OBJECTIVE: To determine if the decision to treat uncomplicated mild hypertension with drugs, in accordance with the WHO/ISH guidelines based on a series of blood pressure (BP) measurements over six months, resulted in the treatment of patients at high risk, on the basis of echocardiography. BACKGROUND: The value of echocardiography in mild hypertension management remains is unclear. METHODS: One hundred and eighteen patients with mild hypertension (90 to 105 mmHg diastolic BP and/or 140 to 180 mmHg systolic BP) were examined by echocardiography at inclusion and followed up for 6 months by a single physician unaware of the echographic results. RESULTS: Drug treatment was given to 48 patients, and 70 remained untreated. Treated patients had higher echographic indices than untreated patients (all p < 0.05): LV mass/body surface area (82.8 +/- 15.9 vs 74.7 +/- 15.0 g/m2), interventricular septal thickness (9.7 +/- 1.7 vs 8.5 +/- 1.3 mm). LV posterior wall thickness (8.4 +/- 1.1 vs 7.8 +/- 1.1 mm), relative wall thickness (0.37 +/- 0.06 vs 0.34 +/- 0.06). Left ventricular (LV) geometry was normal in 98 patients, and 20 had LV concentric remodeling. The 10-year coronary disease risk (Framingham equation) was higher in treated patients than in untreated patients (10.0% vs 6.3%; p < 0.002), and in the 20 patients with concentric remodeling than in those with normal LV geometry (10.4%) vs 4.2%; p < 0.005). Nine of these 20 patients were still untreated at the end of the six month follow-up period. CONCLUSION: Rigorous application of the WHO-ISH clinical guidelines in a group of mild hypertensive patients, led to the treatment of patients with slightly higher LV mass and more concentric LV geometry than were found in those not treated. However, a high-risk subgroup, with concentric remodeling, was not identified and left untreated. PMID- 9749138 TI - [Echocardiographic assessment of the left ventricular fractional shortening/end systolic stress relation in untreated hypertensive patients]. AB - The present study was designed to assess the left ventricular (LV) function by the fractional shortening (FS)/end-systolic stress (ESS) relation in untreated hypertensive patients. METHODS: Two patient groups were examined: group 1 was made up of 50 patients with essential mild hypertension (44 +/- 14 years) and group 2 consisted of 16 normotensives (35 +/- 12 years). LV mass, endocardial FS, mid-wall FS and meridional ESS were measured according to the classic formulas, as well as the 24-hour ambulatory blood pressure (ABP). FS/ESS relation was analysed in hypertensive patients by 2 different ways: 1 degree graphically, with the 95% confidence interval of the normal FS/ESS relation as a reference: 2 degrees statistically, by comparing the observed FS values with those FS value expected from ESS, using equations derived from normal subjects (McMemar chi 2 test, or binomial rule). RESULTS: The graphic examination allowed for the identification of a depressed, normal or increased LV function in, respectively, 7, 31 and 12 patients, using the endocardial FS/ESS relation, against 30, 18 and 2 patients, using the mid-wall FS/ESS relation. Likewise, the statistical analysis of the differences between the observed FS values and the expected FS values showed 13 discrepancies, all of them being related to an overestimation of the LV function, an assessed by the endocardial FS/ESS relation (p < 0.01). Such a trend turned out to be more significant in patients with an increased ABP (n = 31; p < 0.05) than in "white coat" hypertensives (n = 9; p < 0.08): in contrast, it was of the same extent in patients with an increased LV mass (n = 17; p < 0.05) as in those with a normal LV mass (n = 33; p < 0.05). CONCLUSIONS: 1/After adjustment for ESS, mid-wall FS (a direct measure of myocardial fiber shortening) was more often decreased than endocardial FS (reflecting chamber dynamics). 2/Mid wall FS was also found to be a better tool for assessing LV function in patients with normal LV mass, and, to a lesser extent, in "white coat" hypertensives. PMID- 9749139 TI - [Effect of reduced arterial compliance on the diastolic function of the heart]. AB - BACKGROUND AND OBJECTIVE: Reduced arterial compliance is associated with age, hypertension and left ventricular hypertrophy. It is unclear if and how reduced arterial compliance affects the diastolic properties of the heart. PATIENT POPULATION: We examined 64 normotensive and 27 hypertensive individuals, (31 females, 60 males). Their ages ranged from 19 to 78 years (mean 43 +/- 14.7 y), BP ranged from 80/50 to 180/110 mmHg. Exclusion criteria were atrial fibrillation, coronary or peripheral vascular disease, diabetes, anemia, hypercholesterolaemia, as well as antihypertensive drug therapy. METHODS: Diastolic function was assessed by Doppler measurement of E and A wave velocities of Mitral flow. E acceleration and deceleration slopes and times. LV mass was calculated from an M- mode echocardiogram. Arterial compliance was assessed by an automatic pulse wave velocity (PWV) measurement. RESULTS: Age and PWV correlated significantly with the mitral E and A waves, the E acceleration time. E deceleration and E deceleration time. No significant correlations were found between SBP, DBP or LV mass and the parameters of diastolic function (see table). PMID- 9749140 TI - [Effects of high sodium intake on ventricular remodeling in mice]. AB - BACKGROUND: Despite extensive research, controversy still exists regarding the role of dietary sodium intake on hypertension and left ventricular (LV) hypertrophy. Echocardiography is a powerful tool to assess LV hypertrophy and recent technical developments allow now its use in small animals. METHODS: We examined the effect of high sodium intake on LV geometry using echocardiography in mice. Three groups of Swiss mice were submitted, for 8 weeks, to different salt diets (0.6, 2 and 4% NaCl; n = 12, n = 8 and n = 12 respectively). LV end diastolic (ED) septal and posterior wall thicknesses, LV ED diameter were measured at baseline, 4 and 8 weeks. RESULTS: At baseline, heart rate, LV ED septal and posterior wall thicknesses and LV ED diameter were similar between groups. At 8 weeks, for similar heart rate, LV ED posterior wall thickness were not different (0.6%: 0.64 +/- 0.01, 2%: 0.62 +/- 0.08 and 4%: 0.67 +/- 0.03 mm respectively) but LV septal wall thickness ass increased in a salt diet dependent manner (0.6%: 0.63 +/- 0.01, 2%: 0.75 +/- 0.01, 4%: 0.80 +/- 0.02 mm, p < 0.01). This increase was correlated with urinary sodium excretion (r = 0.84, p < 0.01) but occurred in the absence of change in arterial pressure (tail-cuff plethysmography; 0.6%: 135 +/- 6.2%: 127 +/- 4 and 4%: 139 +/- 9 mmHg respectively). The in-vivo interventricular septal remodeling was confirmed in perfused fixed preparations of hearts. CONCLUSION: Echocardiography allows precise measurements of regional LV wall dimensions in mice, and high sodium intake, in the absence of hypertension, induces interventricular septal remodeling. PMID- 9749141 TI - [Role of B1 receptors in the endothelial protective effect of ischaemic preconditioning]. AB - The aim of this study was to assess whether the cardioprotective effect of ischaemic preconditioning (IPC) on endothelial function in resistance coronary arteries of the rat involves activation of kinin receptors. Isolated rat hearts perfused under constant flow conditions were exposed to 30 min of partial ischaemia (flow rate 1 mL/min) followed by 20 min of reperfusion. Preconditioning was performed with 5 min zero-flow ischaemia and 10 min reperfusion before the 30 min ischaemia. After the 20-min reperfusion period, coronaries were precontracted with U-46619 0.1 microM, and the coronary response to the endothelium-dependent vasodilator, serotonin (5-HT, 10 microM), was compared to that of the endothelium independent vasodilator, sodium nitroprusside (SNP, 3 microM). Kinin B1 and B2 receptors were blocked with perfusion of either [Lys0, Leu8, des-Arg0]-Bradykinin 30nM (LLDBK) or Hoe 140 10 nM (Hoe) respectively, started 15 min before IPC or a corresponding sham period and stopped just before the 20-min reperfusion period. In untreated hearts, ischemia diminished selectively 5-HT-induced vasodilatation, compared to sham hearts (without ischaemia). The vasodilatation by SNP was unaffected after ischaemia and reperfusion. Preconditioning in untreated hearts preserved the vasodilatation produced by 5-HT. Treatment of hearts with either Hoe or LLDBK had no effect on the vasodilatation produced by both 5-HT and SNP in sham hearts. Pre-treatment with Hoe did not block the protective effect of IPC on the 5-HT vasodilatation. LLDBK halved the protective effect of IPC on endothelium dependent vasodilatation. In addition, the protective effect of BK on the endothelial function in the isolated rat heart was blocked by LLDBK. These results suggest that IPC and exogenous kinin perfusions afford protection to endothelial function in resistance coronary arteries of the rat partially by activation of B1 kinin receptors. B2 receptors do not play any role in that protection. PMID- 9749142 TI - [Suprasystolic dP/dt max an additional parameter of contractile cardiac function obtained by cuff oscillometric tracings]. AB - Technological evolution allowed to record high fidelity traces that--when analysed by complex mathematical systems--may provide extremely detailed and new information about all the factors involved in the determinism of pulse wave. Suprasystolic waves, i.e. those recorded immediately before systolic pressure, may be regarded as similar to aortic pressure waves evaluated during cardiac catheterization. Suprasystolic dP/dt max was calculated from the profile of pulse wave recorded by the DynaPulse, an automatic portable non-invasive oscillometric method to simultaneously measure BP and analyse arterial waveforms, in 10 normal healthy subjects (age 37 +/- 5) and 5 subjects with ischaemic dilatative cardiomyopathy (age 41 +/- 7) whose ejection fraction--invasively assessed--was < 40%. The 24 h dP/dt max curves were analysed by parametric and non parametric tests. We found a significant difference (p < 0.001) in the average 24-h dP/dt max between healthy subjects (471 +/- 36.7 mmHg/sec) and patients with impaired cardiac function (271 +/- 54.2 mmHg/sec). The average daytime and nighttime dP/dt max values showed significantly higher values in normal subject in comparison to patients with heart failure (daytime 7.23 h: 529 +/- 74 mmHg/s versus 227 +/- 64 mmHg/s, p < 0.001; nighttime: 572 +/- 82 mmHg/s versus 202 +/- 67 mmHg/s, p < 0.001). We also found a difference in the occurrence of acrophases, at similar blood pressure value, i.e. the highest dP/dt values occurred during the night in normal subject, the opposite in ischaemic patients. Furthermore, the dP/dt max correlates only with systolic blood pressure. PMID- 9749143 TI - [Models of arterial pressure using a Windkessel type model: role of the functional arterial properties]. AB - The Windkessel model is a linear model which does not take into account the structural and functional variations of the arteries related to the pulsatility of the blood pressure (BP) and its variations between systole and diastole. OBJECTIVE: To analyse the performance of a BP modelisation where the parameters of AC are adjusted in a dynamic fashion according to a curvilinear relationship of the arterial properties (compliance) in relationship to the BP between systole and diastole. DESIGN AND METHODS: 9 control subjects (age 25 +/- 3). The non invasive measures of the radial BP waveform (Millar tonometry) was compared to that constructed by an electric simulator reproducing the model in a sysmetrical network subdivised into 121 segments where we introduced for each subject: at cardiac level, the aortic stroke volume (Doppler echocardiography), and at the radial level, the dynamic values of compliance and diameter according to an arc tangent model (measured by arterial echography NiUS02). RESULTS: The BP obtained by the adjusted model, where the AC parameter follows the curvilinear, relationship dV/dP measured experimentally, was not significantly different from the experimental, while in the constant model (AC at mean BP level) the systolic BP was different. CONCLUSION: This work shows in an experimental way the limits inherent in simplification in the Windkessel modelisation of the vascular system with constant parameters. It shows in a conduction artery the influence of the functional properties of the arterial wall on the level of systolic and diastolic BP. PMID- 9749144 TI - [Cardiovascular risk factors and shift work in men living in Haute-Garonne, France]. AB - OBJECTIVE: The goal of this study was to examine the relationship between shift work and cardiovascular risk factors in a cross sectional survey. MATERIAL AND METHODS: 2 610 men of the PRIME study, aged 50-60 years, residing in Haute Garonne (France) and selected from health centers and various firms were screened. White-collar workers, retired and disabled men at the time of the study were excluded so the analysis was performed on a sample of 1,341 subjects. Work was categorized as day work (n = 1,161) and three types of shift work: 2 x 8 (n = 108), 3 x 8 (n = 41) and night (n = 31). A comparison of cardiovascular risk factors was performed in the different types of work. RESULTS: Univariate analysis showed that systolic blood pressure was significantly higher among 2 x 8 workers (132.7 +/- 17.4 mmHg) than in other shift workers and day workers (126.9 +/- 15.1 mmHg). The same difference was observed for diastolic blood pressure. Plasma triglycerides were significantly higher among night workers (1.60 +/- 0.76 g/l) than among day workers (1.29 +/- 0.81 g/L). Logistic regression analysis showed that the risk of hypertension, defined as systolic (> or = 140 mmHg) and diastolic (> or = 90 mmHg) blood pressure and/or antihypertensive drug, was 2 times higher among 2 x 8 shift workers (OR: 2.14, 95% CI [1.42-3.23]) than among day workers after adjustment for BMI, alcohol and tobacco consumption and plasma triglyceride levels. The same result (OR: 1.99, 95% CI [1.28-3.12] was obtained with a different definition of hypertension (systolic > or = 160 mmHg and/or diastolic > or = 95 mmHg blood pressure and/or antihypertensive drug). Among night workers the risk of triglycerides > 2 g/L was higher (OR: 2.52, 95% CI [1.06-6.01] than among day workers. The risk of both hypertension (140/90 mmHg) and hypertriglyceridemia in comparison to day workers was (OR: 2.01, 95% CI [1.33 3.03]) and (OR: 2.44, 95% CI [1.14-5.21]) for 2 x 8 shift workers and night shift workers respectively. CONCLUSION: The results of this study, putting into evidence a relationship between shift work, hypertension and high level of triglycerides, justify a careful screening by occupational physicians of cardiovascular risk factors among shift workers. PMID- 9749145 TI - [Ankle-arm blood pressure index (AABPI) in hemodialysis patients]. AB - AABPI, calculated as the ratio of systolic ankle/systolic arm blood pressure, has been recently found to be a strong predictor of cardiovascular and overall mortality in hemodialysis patients. The aim of our study was to confirm the role of this test in dialysis patients, a population with high prevalence of vascular diseases. Two hundred and twenty-six patients were studied, of which the AABPI could be measured in 217. There were 134 males (61%) and 83 females (39%) with a mean age of 61.3 +/- 17.4 years. The mean AABPI for the studied patients was 1.02 +/- 0.26; a past history of coronary artery (CAD), and/or cerebrovascular (CVD), and/or peripheral vascular disease (PVD) was present in 97 (45%) of these patients. This latter group had a mean AABPI less than controls with any vascular diseases (0.9 vs 1.1 p < 0.0001). For patients with or without CAD and PVD, the AABPI was respectively 0.84 +/- 0.3 vs 1.06 +/- 0.23 (p < 0.001) and 0.91 +/- 0.28 vs 1.08 +/- 0.22 (p < 0.001). In the group of patients with CAD, CAV, and PVD the positive and negative predictive value of AABPI was respectively of 66 and 74%. Diabetes was associated with a significantly lower AABPI (p < 0.02), gender did not influence AABPI. Significant positive correlation was found between AABPI and age (r2 = 0.46; p < 0.01). For patients with CAD, CVD and/or PVD no differences were found in serum lipid parameters (triglycerides, cholesterol, HDL-cholesterol, and lipoprotein a). Cumulative survival curves showed a lower mortality in patients with AABPI. 0.9 (Logrank test p < 0.001). We confirm that AABPI is a powerful non-invasive marker for the presence of systemic atherosclerotic disease in hemodialysis patients. PMID- 9749146 TI - [Do WHO-ISH guidelines identify high risk mild hypertensive patients?]. AB - OBJECTIVE: To assess the ability of 1993 WHO-ISH recommendations to identify patients who need drug treatment. METHODS: 268 hypertensive patients with suspected mild hypertension were preselected for this study at their first visit at the referral center. 123 were included after a short in-hospital work-up when they fulfilled the 1993 WHO-ISH criteria for mild hypertension (90-105 mmHg diastolic BP and/or 140-180 mmHg systolic BP). Echocardiography was performed in all patients by the same investigator according to ASE convention. The combined 10-year risk of stroke and coronary heart disease was calculated with the Framingham equation. Patients were then followed up for six months by the same physician blinded to echographic results and risk calculations and applying the WHO-ISH guidelines (monthly BP measurement and subjective assessment of risk). Five patients were excluded, for reasons unrelated to the protocol. RESULTS: The decision of drug treatment was taken at the 1st, 2nd, 3rd, 4th, 5th, 6th monthly visit after work-up in 2, 6, 25, 7, 2 and 6 patients, respectively. Among these 118 patients, 48 patients (29 male, 19 female) were eventually treated and 70 (49 male, 30 female) remained untreated. BP s at preselection and on a day of work-up were similar in both groups. Patients in whom drug treatment was prescribed were older and had higher lef ventricular mass (LVM) than untreated patients, but only 2 of them (all in the treated group) had LVM values above usual thresholds (LVM > 125 g/m2, in men and women). Stroke and coronary risks were both higher in treated than in untreated patients (p < 0.05). The physician using the guidelines decided to treat only 19 of the 38 patients with a 10-year risk < 10% (true positive), whereas she decided to treat with drugs 12 patients among the 44 with a 10-year risk < 5%. CONCLUSION: The difference in LVM between untreated and treated patients support the validity of the WHO-ISH guidelines, but the measurement of LVM did not bring much information for managing the individual patient. Application of these guidelines did not satisfactorily identify high risk patients and could lead to over-treatment of low risk patients. PMID- 9749147 TI - [Frequency of improper diagnosis of hypertension]. AB - The aim of this work is to estimate the real prevalence of hypertension in a population declared hypertensive by general practitioners. This prospective study has lasted 30 months from October 1994 to March 1997). It has concerned 2,151 patients that had been declared hypertensive by general practitioners. Each patient had 2 visits at 15 days interval. During each visit the blood pressure (BP) is measured 4 times at 5 min interval with a mercury tensiometer. The patient remains supine for 20 min. Patients declared normotensive on these 8 measurements are controlled once every 6 months. Those that are declared hypertensive are distributed in 2 groups: the patients that have a systolic blood pressure (SBP) superior to 160 mm of mercury (mmHg) and/or diastolic blood pressure (DBP) superior to 95 mmHg are treated with the habitual follow-up. Those that have a SBP and DBP between 140-160 mmHg and 90-95 mmHg have a control visit at the end of the first and the third month. After this period patients whose BP remains between 140-160 mmHg and 90-95 mmHg have an ambulatory blood pressure measurement (ABPM). This study included 1,635 women and 516 men. Average age was 54 +/- 11 years. The average of 8 measurements of SBP/DBP was 148.3 +/- 22.5/93 +/- 13 mmHg. After 8 measurements, out of 2,151 patients, 37.8% (841 patients) were declared normotensive and 62.2% (1,337 patients) had a SBP and/or DBP > 140/90 mmHg. This group was controlled after one month and 3 months. A subgroup (254 patients) was declared normotensive, and the 588 patients whose BP remained between 140-160 mmHg and 90-95 mmHg had an ABPM : 481 (48%) were then declared normotensive and 307 (52%) were declared normotensive patients. After repeated controls, with the use of ABPM, only 37.3% were truly hypertensive. This study confirms that a long follow-up is necessary before labelling a patients as hypertensive. BP can remain abnormal for weeks and months. A large group of "hypertensive" patients are normotensive people with a white coat effect or so labelled because of errors in technique or the absence of a sufficient delay so that BP returns to normal. PMID- 9749148 TI - [A software for comparing ambulatory blood pressure measurements (ABPM)]. AB - The software (MAPA-PC) for personal computers, designed by the author, provides precise and fast studies of ABPM, especially when comparing two anti-hypertensive treatments. It implies strict definitions of groups (e.g. dippers), indices (peak, trough, peak/trough ratio), and the type of smoothing of measured values: means (over 1, 2 or 3 hours) or smoothing by functions (polynomials or trigonometric series). The software calculates standard parameters (nighttime and daytime means, peak, trough...) as well as more original values (nighttime and period from points of inflexion in the curve, slope in these points, coefficients of variation on differences between 2 ABPM of a given subject...). It uses statistical tests (Mann and Whitney's U. Pearson's chi square, Snedecor's F) to compare results of treatment. It provides many types of curves (blood pressure lines, box-plots) which show kinetics and variability of blood pressure. The user can compare various types of smoothing (e.g. 2 vs 3 harmonics in Fourier series). The software was used on 237 records of 79 patients, in a randomized trial. It was written with Turbo Pascal and works on DOS personal computers. It is interesting for epidemiologic studies or clinical trials in which ABPM are used. PMID- 9749149 TI - [A quantitative analysis of a predictive model of ambulatory blood pressure monitoring integrating physical activity recording]. AB - OBJECTIVE: To determine how much of the variations of blood pressure during a 24 hour period could be accounted for by a change in activity using an accelerometer to detect the physical activity and establish a predictive model. MATERIALS AND METHODS: 18 healthy subjects (mean age 25 +/- 2 yrs) were studied during daily life (24 hours) twice one week apart. The systolic and diastolic blood pressure, heart rate (HR), and time of measure were recorded by ambulatory monitoring using Spacelabs (4 measures per hour). A portable digital memory device was designed for the 24 hours ambulatory monitoring of HR (ECG) and physical activity. This device consists of an ECG Holter (ELA medical model Cinesis with digital memory) and a three piezoresistive type accelerometer sensors (prototype ELA research) able to record physical activity in the 3 space dimension. RESULTS: The data of the first recording were compared to the predicated values from the application of a logarithmic model of activity to the second recording. The model then predicted 53 +/- 19% of the systolic BP values of the test day. The mean individual difference for a given time period of one hour between the measured and the predicted systolic BP from the model was 1.45 +/- 3.1 mmHg with a range of [-6.9; 3.4 mmHg]. The mean individual systolic BP difference for the same given time period of one hour but without predictive model was 1.29 +/- 10 mmHg with a range of [-28; 43 mmHg]. CONCLUSION: This study show that 3 D accelerometer is an easy tool to program individual model of ambulatory blood pressure variability. The introduction of this qualitative method seems logical in therapeutic trial. PMID- 9749150 TI - [Ambulatory blood pressure, intima media thickness, global cardiovascular risk and therapeutic decisions]. AB - OBJECTIVE: To determine the influence of ambulatory blood pressure monitoring (ABPM), carotid intima media thickness (IMT) and global cardiovascular risk on the therapeutic strategies issued from our hypertension unit. METHODS: All essential uncomplicated and never treated hypertensive patients referred to our hypertension unit between 1996 and 1997 for etiologic or target organ damage evaluation were considered eligible. We excluded diabetics and patients with renal disease who need a specific therapeutical approach. 54 patients (44.7 +/- 10.1 years) were included (40 men). All patients underwent an ABPM measurement. The right common carotid IMT measurement had been performed (0.06 +/- 09 mm). The global cardiovascular risks were assessed with the Framingham prediction chart taking into account age, sex, total cholesterol, smoking status and systolic office blood pressure. According to the therapeutic decision three groups were made up: group 1 lifestyle counselling (n = 13), group 2 single drug therapy (n = 31), and group 3 combination therapy (n = 10). RESULTS: No significant difference was found in age, sex ratio, prevalence of severe hypertension, office systolic blood pressure, body mass index, global cardiovascular risk between the three groups. In contrast ABP (24 h ABP mmHg: group 1: 128.23 +/- 6.91/79.7 +/- 6.4; group 2: 140.48 +/- 9.7/97.48 +/- 8.17; group 3: 152.4 +/- 15.35/99.4 +/- 12.14 p < .0001) and IMT (group 1: 10.55 +/- .09, group 2: .59 +/- .07, group 3: .66 +/- .11 p = .02), were significantly higher in group 3 than in group 2 and in group 1. The percentage of white coat hypertensives was higher in group 1 than in group 3 (group 1: 61.5%, group 2: 3.2%, group 3: 0%). In the whole population, the higher was the global cardiovascular risk, the higher was the common carotid intima media thickness. In this study the global cardiovascular risks are not related to therapeutic decisions. Therapeutic strategies are influenced by ABP level and by the vascular remodeling which depends partly on the global cardiovascular risk. PMID- 9749151 TI - [Arterial hypertension and socioeconomic status in a representative population of French railway drivers]. AB - High blood pressure (HDP) is known as a cardiovascular risk factor depending both on environmental and socio-economic factors. METHODS: From October 1993 to october 1994 a cross sectional study was carried out among 1,855 French railway drivers (FRD) representative of the 17,432 males FRD, aged 25 to 54 years. Age, weight, height, hip and waist, smoking, living area, type of train they drove (goods, suburban, TGV, inter-city trains), their grade (3 grades) were recorded. HBP corresponds to systolic blood pressure (BP) 160 mmHg or diastolic BP > or = 95 mmHg or to normal BP under antihypertensive medication. The analysis was carried out according to 2 age classes: 20-36/37-54 years. RESULTS: In our sample 8.5% of FRD suffered from HBP, 3.5% in the younger class, and 13.5% in the other one. Using univariate analysis, among the oldest, subjects with lower grades suffered more often from HBP (19%). Paris area was more often related to HBP for the 20-36 years (6.7%). This was the case for the oldest living in Paris area (19.7%), and the North East (15.2%). Subjects with central obesity (19.9 vs 6%), and hyperlipidemic FRD (20.9 vs 10.9%) were more often related to HBP in the 35 54 years group Whatever the type of train they drove no difference was found. In multivariate analysis, (stepwise logistic regression BMDP LR) independent HBP factors are: age OR 3.4 IC 1.9-5.9 (20-36 vs 37-54), central obesity OR 1.7 IC 1.1-2.6, tobacco consumption OR 2.1 IC 1.2-3.5 (smokers vs non smokers), ex smokers: OR 2.3 IC 1.3-3.9 (Ex-smokers vs non smokers), living area (all regions vs South East in the Mediterranean border). Nevertheless, grades and type of train they drove were not independent factors. CONCLUSION: These results show the determining part played by environmental factors: age, central obesity, living area and tobacco consumption in the determinism of HBP in professional background: these factors can account for the difference observed in professional factor (grade). PMID- 9749152 TI - [Variations of blood pressure during the month of Ramadan]. AB - The goal of this work is to study the consequences of the last on variations of the blood pressure (BP) in the course of 24 hours. From 1994 to 1997 we have selected 99 hypertensive patients and studied their BP profile. This study included 72 women and 27 men. Their age varies from 22 to 72 years (average 56.7 +/- 9 years). All these patients has an ambulatory blood pressure measurement (ABPM) before the fast and during Ramadan. Before Ramadan the period of the sleep goes from 10 pm +/- 1 h to 8 am +/- 1 h. During the month of Ramadan, the sleep lasts from 0 h +/- 1 to 9 am +/- 1 h. [table: see text] No statistically significant difference is noted between these 2 periods neither for the systolic BP (SBP) nor for the diastolic BP (DBP), for the BP of 24 hours, and the diurnal and nocturnal periods. We have then the compared the hourly average on 24 hours of the 99 patients. We observed that during the month of Ramadan the peak of the awakening is delayed by 2 hours and the nocturnal through is delayed by 1 hour. After this study, which is the first one to deal with variations of blood pressure during the fast of Ramadan we can confirm that in patients with essential hypertension without complications, the fast is well supported. The variations of BP are minimal and are related to the variations of the sleep, activity and eating pattern. PMID- 9749153 TI - [Twenty-four hour time and frequency domain variability of systolic blood pressure and heart rate in an experimental model of arterial hypertension plus obesity]. AB - Modifications of heart rate (HR) and systolic blood pressure (SBP) variabilities (V) have been reported in the human syndrome arterial hypertension plus insulin resistance. The aim of this study was to characterize the 24 h SBPV and HRV in both time and frequency domains during weight increase in dogs fed ad libitum with a high fat diet. Implantable transmitter units for measurement of blood pressure and heart rate were surgically implanted in five beagle male dogs. BP and HR were continuously recorded using telemetric measurements during 24 hours, before and after 6 and 9 weeks of hypercaloric diet in quiet animals submitted to a 12h light-dark cycle. To study nychtemeral cycle of SBP and HR, two periods were chosen: day (from 6.00 h to 19.00 h) and night (from 23.00 h to 6.00 h). Spontaneous baroreflex efficiency was measured using the sequence method. Spectral variability of HR and SBP was analyzed using a fast Fourier transformation on 512 consecutive values and normalized units of low (LF: 50-150 mHz, reflecting sympathetic activity) and high (HF: respiratory rate +/- 50 mHz, reflecting parasympathetic activity) frequency bands were calculated. The energy of total spectrum (from 0.004 to 1 Hz) was also studied. Body weight (12.4 +/- 0.9 vs 14.9 +/- 0.9 kg, p < 0.05). SBP (132 +/- 1 vs 147 +/- 1 mmHg, p < 0.05) significantly increased after 9 weeks of hypercaloric diet. A nycthemeral HR rhythm was present at baseline (day: 79 +/- 1 vs night: 71 +/- 1 bpm) but not after 9 weeks (day: 91 +/- 4 bpm ; night: 86 +/- 2 bpm). Concomitantly, the efficiency of spontaneous baroreflex decreased at 6 weeks (36 +/- 1 vs 42 +/- 2 mmHg/ms, p < 0.05). A significant decrease in HF energy of HRV was found after 6 but not after 9 weeks. LF energy of SBPV was increased at 6 but not at 9 weeks (table). [table: see text] In conclusion, this study shows that an hyperlipidic and hypercaloric diet induces transient variations in autonomic nervous system activity which could be the physiopathological link between obesity, insulin resistance and arterial hypertension. PMID- 9749154 TI - [Autonomic contribution to the blood pressure and heart rate variability changes in early experimental hyperthyroidism]. AB - A great deal of uncertainty persists regarding the exact nature of the interaction between autonomic nervous activity and thyroid hormones in the control of heart rate (HR) and blood pressure (BP). Thyrotoxicosis was produced by a daily intraperitoneal (i.p.) injection of L-thyroxine (0.5 mg/kg body wt in 1 ml of 5 mM NaOH for 5 days). Control rats received i.p. daily injections of the thyroxine solvant. Autonomic blockers were administered intravenously: atropine (0.5 mg/kg), atenolol (1 mg/kg), atenolol + atropine or prazosin (1 mg/kg). Eight animals were studied in each group. Thyroxine treatment was sufficient to induce a significant degree of tachycardia (423 +/- 6 vs 353 +/- 4 bpm; p < 0.001, unpaired Student's tests), systolic BP elevation (142 +/- 3 vs 127 +/- 2 mmHg; p < 0.001), pulse pressure increase (51 +/- 2 vs 41 +/- 2 mmHg, p < 0.01), cardiac hypertrophy (1.165 +/- 0.017 vs 1.006 +/- 0.012 g, p < 0.001), weight loss (-21 +/- 2 g; p < 0.001) and hyperthermia (37.8 +/- 0.1 vs 37.0 +/- 0.1 degrees C, p < 0.001). The intrinsic HR observed after double blockade (atenolol + atropine) was markedly increased after treatment with thyroxine (497 +/- 16 vs 373 +/- 10 bpm, p < 0.05). Vagal tone (difference between HR obtained after atenolol and intrinsic HR) was positively linearly related to intrinsic HR (r = 0.84; p < 0.01). Atenolol neither modified HR nor BP variability in rats with hyperthyrodism. The thyrotoxicosis was associated with a reduction of the 0.4 Hz component of BP variability (analyses on 102.4 sec segments, modulus 1.10 +/- 0.07 vs 1.41 +/- 0.06 mmHg; p < 0.01). Prazosin was without effect on this 0.4 Hz component in these animals. These data show a functional diminution of the vascular and cardiac sympathetic tone in experimental hyperthyroidism. Increased intrinsic HR resulting from the direct effect of thyroid hormone on the sinoatrial node is the main determinant of a tachycardia leading to a subsequent rise in cardiac output. The resulting BP elevation could reflexly induce a vagal activation and a sympathetic (vascular and cardiac) inhibition. PMID- 9749155 TI - [Brain mineralocorticoid receptor and blood pressure control in the conscious normotensive rat]. AB - Brain control of arterial blood pressure in man and in animals has been studied increasingly over the last few decades. Despite our knowledge about short term regulation (chemoreceptor and baroreceptor reflexes) there is much more uncertainty about the degree of involvement of brain mechanisms in long term control of blood pressure, and in hypertension. The last decade, a role of brain mineralocorticoid receptors (MR) has been outlined in animal experiments. Stimulation of brain MR by aldosterone or related mineralocorticoids induces an increase in blood pressure and hypertension under chronic conditions. These effects of mineralocorticoid excess can be blocked by specific MR antagonists administered centrally. Stimulation of brain glucocorticoid receptors, as compared to stimulation of brain MR, has an opposite effect, i.e. decreases blood pressure. As in the typical peripheral target organs of aldosterone, in the brain, enzymatic protection of MR against glucocorticoids appears to play an important role. We showed that in conscious normotensive rats intracerebroventricular (i.c.v.) injection of a specific MR antagonist (RU28318) in a low dose (10 ng) decreases blood pressure by about 20% without affecting heart rate. A similar but smaller effect (not statistically significant) was observed in conscious female rats. Only in the male rats an increased diuresis occurred and this may have contributed to the observed hypotension. We conclude that hypertension caused by mineralocorticoid excess may depend on a concerted action of the steroid on the kidney and on the brain. The mechanism by which brain MR increases blood pressure is unknown. It is possible that increased sympathetic outflow and renal mechanisms are involved. Interference with brain MR not only affects blood pressure but it also has effect on renal function. PMID- 9749156 TI - [Effects of hydroxyl radicals on purified angiotensin I converting enzyme]. AB - Somatic angiotensin-converting enzyme (ACE) is a protein which contains two similar domains (N and C), each possessing a functional active site. The relationship between ACE, its natural substrates and oxygen free radicals is starting to be explored. On one hand, superoxide anions production is induced by angiotensin II and on the other hand, activated polynuclear neutrophils, through free radicals generation, alter endothelial ACE activity. In this study, we examined the impact of hydroxyl radicals (.OH) on purified ACE. .OH were produced using a generator: 2,2'-azo-bis 2-amidinopropane (GRH) provided by Lara-Spiral (Fr). GRH (3 mM), in a time-dependent fashion, inhibited ACE activity. When ACE was co-incubated for 4 h with GRH, its activity decreased by 70%. Addition of dimethylthiourea (DMTU: 0.03 to 1 mM) or mannitol + methionine (20/10 mM), two sets of .OH scavengers, produced a dose-dependent protection on ACE activity. To examine whether oxidation of thiol groups in the ACE molecule could be involved in the action of GRH, the effects of thiol reducing agents: mercaptoethanol and dithiotreitol (DTT) were investigated. These compounds produced a dose-dependent and significant protection; with 100% protection at 0.2 and 0.3 mM for mercaptoethanol and at 0.1 mM for DTT. The hydrolysis of two natural and domain specific substrates were also explored. The hydrolysis of angiotensin I preferentially cleaved by the C domain was significantly (p < 0.01) inhibited by 57, 58 and 69% in contact with 0.3, 1 and 3 mM GRH [in nmol angio II formed/min/nmol of ACE, n = 4; 35.9 +/- 0.6 (control), 15.5 +/- 2.8 (GRH : 0.3 mM), 15.1 +/- 0.5 (1), 10.9 +/- 0.6 (3)]. The hydrolysis of the hemoregulatory peptide (hp), preferential substrate for the N domain was not affected by GRH at 0.3 mM and inhibited by 28% (not significant) by 1 mM GRH [in nmol ph hydrolized/min/nmol ACE, n = 4; 12.6 +/- 1.9 (control), 14.9 (GRH : 0.3 mM), 8.3 +/- 4.0 (1). These results demonstrated that .OH affect ACE activity and could suggest a privileged impact of GRH on the C domain. The precise sites of action of .OH remain unknown. The Cys residues near the active centers, by forming disulphide bridges during the oxidation could be of critical importance. Further studies will be needed to determine whether oxidative stress again ACE can be involved in the genesis of inflammatory vascular pathologies. PMID- 9749157 TI - [Functional decoupling of left ventricular beta-adrenoceptor in a canine model of obesity-hypertension]. AB - OBJECTIVE: To assess cardiac beta-adrenoceptors (beta-AR) in an obesity hypertension model. METHODS: Six male beagle dogs (aged 35 +/- 5 months) receiving during 30 weeks a high-fat diet with 60% uncooked beef fat were compared to 6 normal beagle dogs. With right auricular and left ventricular samples we analysed cardiac beta-AR density through binding study using [125I] cyanopindolol. beta 1 and beta 2 densities were obtained by competition with CGP 20712A. Affinity state of beta-AR was assessed by competition with isoproterenol. Noradrenaline plasma level was assayed by HPLC. Left ventricular mass (LV mass) was measured by echocardiography. Results are expressed as mean +/- SE. All comparisons were performed using a variance analysis (*: p < 0.05). RESULTS: Systolic blood pressure was significantly higher in obesses (245 +/- 8 vs 197 +/- 10 mmHg in controls). Diastolic blood pressure did not differed between both groups (93 +/- 3 vs 84 +/- 3 mmHg in controls). Noradrenaline plasma levels were similar in both groups (276 +/- 30 vs 235 +/- 50 pg/mL in controls). Obesses were characterized by higher LV mass (80 +/- 24 vs 67 +/- 15 g in controls*). Right auricular and left ventricular beta-AR densities were not different in obesses (57 +/- 6 and 67 +/- 4 fmoles/mg protein) and in controls (68 +/- 7 and 63 +/- 9 fmoles/mg protein). The beta 1-AR proportion was the same in obesses and controls in right auricule (63 +/- 4 vs 64 +/- 3% in controls) and left ventricule (59 +/- 3 vs 60 +/- 4% in controls). The proportion of beta-AR receptors in a high affinity state was similar in right auricular samples (69 +/- 4 vs 67 +/- 3%) in controls) but was significantly different in left ventricule (28 +/- 6 vs 74 +/- 6%) in controls). CONCLUSION: Left ventricular beta-adrenoceptors came under a specific desensibilisation independent of plasma noradrenaline levels. This functional decoupling of beta-adrenoceptors may account for the progressive systolic dysfunction of hypertensive cardiomyopathy. PMID- 9749158 TI - [Hypotensive effect of endothelin-1 in a rat model of pre-eclampsia]. AB - Hypertensive pregnant rats with inhibition of NO synthase are frequently considered as model of pre-eclampsia with proteinuria, hypertension and elevated endothelin (ET-1) blood levels. We describe here the cardiovascular in vivo effects of ET-1 in this rat model since ET-1 and NO are both important vasoactive mediators in uteroplacental circulation. From day 13 of gestation 2 groups of Wistar female rats were fed control (C) or nitroarginine enriched diet (0.063%, Treated: T). On gestational day 20 mean arterial pressure (MAP, mmHg) was measured via a carotid catheter in pentobarbital (60 mg/kg) anesthetized rats. After chronic NO synthase inhibition hypertension develops; MAP on day 20: 158 +/ 2.2 in T and 113 +/- 2.2 in C, p < 0.001. ET-1 bolus injection (0.1 nmol/kg) is rapidly followed by a decrease in blood pressure significantly more important in T: -46 +/- 5.1 than in C: -30 +/- 2.2. In vivo depressor effect is blocked by the specific antagonist BQ-788. After inhibition of cycloxygenase with acetylsalicylic acid (27 mumol/kg, 30 min before) the hypotension is not modified. Since NO and PGI2 productions are not expected in our conditions, vasodepressor effect can be explained by an endothelial hyperpolarazing factor (EDHF). In conclusion in vivo ET-1 hypotensive effects in pregnant rats are mediated by ETB receptors and more pronounced in hypertensive NO-deprived animals. PMID- 9749159 TI - [Evaluation of the effects of experimental hypertension on monocyte-endothelial cell interactions]. AB - Adhesion of monocytes to endothelial cells is considered as one of the initial factors leading on the long term to the development of atherosclerosis. We evaluated whether hypertension affects adhesion of monocytes on rat carotid endothelium, and whether this adhesion may be modified by a chronic treatment with L-arginine, the physiological precursor of nitric oxide (NO). Hypertension was induced in Dahl rats using a sodium-rich diet (8%), in the absence or the presence of L-arginine (1.25 mg/kg/day). After 1 month, the carotid arteries were isolated, opened longitudinally, and incubated in the presence of monocytes previously rendered fluorescent by incubation with tetramethyl rhodamine isothiocyanate (TRITC), and adherent cells were counted under fluorescence microscopy. Monocyte adhesion was minimal in carotid arteries isolated from normotensive rats (13 +/- 5, n = 8). Hypertension induced a marked, significant increase in monocyte adhesion (97 +/- 17; n = 10; p < 0.01 vs normotensive). This increased adhesion was significantly reduced by chronic treatment with L-arginine (37 +/- 13; n = 12, p < 0.05 vs untreated hypertensive). Thus, hypertension was associated with an increased adhesion of monocytes, which is probably due to a decrease production of NO. The increased adhesion was partly prevented by L arginine, possibly secondary to an increased production of NO. Such an increased adhesion of monocytes may contribute to the increased cardiovascular risk in hypertension. PMID- 9749160 TI - [Effect of chronic bradykinin infusion on angiotensin II hypertension in rats]. AB - In previous studies, we demonstrated that in ANG II-treated rats, prevention of cardiac hypertrophy (CH) by enalapril was blunted by bradykinin (BK) blockade by Hoe140. The putative role of BK was assessed by chronic exogenous BK infusion and in 46 male Sprague-Dawley rats infused with ANG II. ANG II (200 ng/kg/min) alone and associated with BK at low (BKlow, 15 ng/kg/day), mid (BKmid, 100 ng/kg/day) and high doses (BKhigh, 100 ng/kg/min) were delivered by Alzet osmotic pumps for 10 days and compared to control animals (Veh). Values of systolic arterial pressure (SAP, mmHg) in conscious rats and heart weight (HW, mg/g bw) at the end of the study are reported below. Results were submitted to ANOVA and are expressed as mean +/- SEM. PMID- 9749161 TI - [Renal alterations in L-NAME hypertension: influence of losartan and bosentan]. AB - The influence of losartan and the endothelin A and B receptor antagonist, bosentan was assessed on the alterations in renal hemodynamic and function as well as urinary albumin excretion (taken as an index of renal lesions) associated with L-NAME hypertension. L-NAME was given for 4 weeks (20 mg/100 mL in the drinking (fluid) followed by a 2-week period of concomitant treatment with L-NAME and losartan or bosentan (30 and 30 mg/kg, gavage). A group of rats received L NAME without additional treatment and a group of rats were not given L-NAME and served as normotensive controls. Systolic arterial pressure (SAP) was measured before L-NAME, and before and every 5 days of losartan or bosentan treatment period. Urinary excretion of albumin (UAlb) was determined before and at the end of treatment period. Under anesthesia, glomerular filtration rate (GFR) and renal plasma flow (RPF) were estimated by the clearance method and the filtration fraction was calculated [FF = 100* (GFR/RPF)]. PMID- 9749162 TI - [The Hypertension Optimal Treatment Study: efficacy and tolerability on the 36th month]. AB - The international, prospective, randomized HOT study was aimed at determining the influence of a targeted BP reduction on cardiovascular morbidity and mortality. Patients were randomly allocated to 3 DBP targets (< 80, < 85, < 90 mmHg). In addition, the impact of a coprescription of aspirin was studied. The BP target had to be reached within 3 months, according to a well-defined strategy : felodipine 5 mg o.d. as a 1st intention drug, 1, 2 or 3 additional drugs, if necessary, on the following steps. BP measurements were made, using an oscillometric automatic device (Hestia). From April 1992 to October 1994, 18,790 patients with an age range 50-80 years, coming from 26 countries, entered the study. The data collected on the 36th month were in agreement with those obtained on the 12th and the 24th months. Baseline DBP was reduced by 21, 23 and 25 mmHg in the 90, 85 and 80 mmHg target groups, respectively. The rate of patients whose DBP reached the target, obviously increased from the 3rd to the 12th month: from 43 to 56%, 60 to 70%, 74 to 83% in the 90, 85 and 80 mmHg, target groups, respectively. From the 2nd to the 3rd year, BP control was further improved, with a slightly higher rate of controlled patients in the elderly (age > 60 y), especially in the 80 mmHg target group. From inclusion to the 3rd month, one-drug treated patients decreased, whereas 2- or 3-drug treated patients increased. Felodipine-treated patients decreased on the 36th month, but remained over 80%. From the 6th to the 36th month, additional prescription of a betablocker or an ACE-inhibitor increased from 36 to 39%, and from 23 to 28%, respectively; moreover, the side-effects rate decreased from 10.5 to 3.6%, with a special decline in ankle edema from 4 to 1%. In conclusion, the BP reduction observed on the 36th month was of the same extent as that observed in the first months. It seems obviously possible to reach a targeted DBP and to maintain it over time, along with a good acceptability of the treatment. Targeted DBP could be more easily achieved in elderly patients, possibly due to a better drug compliance. PMID- 9749164 TI - [Influence of global cardiovascular risk assessment on the management of hypertension in southwestern France]. AB - 1,160 subjects aged 35-64 years were recruited by the Haute-Garonne MONICA center, and selected by stratified randomization on age and size of home area. The hypertensive group included: 176 subjects newly diagnosed as hypertensives (blood pressure > or = 140/90 mmHg), 86 known but untreated hypertensives and 178 hypertensives under treatment. For each subject a score of coronary heart disease risk based on the Framingham point score probability algorithm was calculated. The prevalence of hypertension was 37.9%. Among the 440 subjects considered as hypertensives, 60% were aware of having hypertension. Only 30% of the 178 patients treated achieved blood pressure control. The population as a whole was at low coronary heart disease risk (< 5% at 10 years); the groups at higher risk were newly diagnosed hypertensives and treated hypertensives. Among known hypertensives, the risk level was higher in treated compared with untreated. In this survey 1) the prevalence of hypertension was high; 2) only 30% of treated hypertensives were below 140/90 mmHg; 3) usual care failed to recognize 40% of hypertensives at same risk level as treated ones; 4) treated hypertensives had higher coronary heart disease risk than untreated known hypertensives. The hypertension therapeutic strategy could be based on the reduction of blood pressure below the threshold 140/90 mmHg rather than on the absolute cardiovascular risk. PMID- 9749163 TI - [Influence of some parameters on the blood pressure reduction under treatment: experience from the Hypertension Optimal Treatment Study]. AB - The HOT study is the largest controlled therapeutic trial conducted to date in hypertension. This international, prospective, randomised trial is designed to determine the optimal blood pressure to be obtained during treatment, in order to achieve optimal reduction of complications and cardiovascular mortality. The HOT study is conducted according to the PROBE methodology (Prospective Randomised Open Blinded Endpoints. It has three objectives: 1) to evaluate the relationship between the development of major cardiovascular events and the DBP target level (DBP < or = 90, DBP < or = 85 or DBP < or = 80 mmHg. 2) to evaluate the relationship between the development of major cardiovascular events and real DBP observed, 3) to determine whether low-dose acetylsalicylic acid (75 mg/day) provides an additional benefit in terms of cardiovascular morbidity and mortality in treated hypertensive subjects. Between April 1992 and October 1994, 18,790 patients, between the ages of 50 and 80 years (26 countries), were randomised to these 3 target DBP groups and several parameters likely to influence the blood pressure fall were identified. The reduction of DBP was all the more pronounced the higher the baseline DBP. For a baseline DBP equal to 100 mmHg, the mean fall was 18 mmHg and for a baseline DBP equal to 110 mmHg, the mean fall was 27 mmHg. Advanced age was also found to be a factor promoting reduction of DBP, which increased after the age of 65 years. Analysed in relation to the type of treatment, this more marked reduction in the elderly showed that monotherapy with a calcium channel blocker was very effective on DBP and especially after the age of 75 years. The data of the study also showed that SBP decreased in parallel to DBP, but to an even greater extent. Thus, a 10 mmHg reduction of DBP induces a reduction of SBP by approximately 20 mmHg. This reduction of SBP related to the level of DBP was even more marked the higher the baseline SBP and the higher the target DBP objective. Thus 80% of patients in the DBP < or = 80 mmHg group had an SBP < or = 150 mmHg during treatment. Among the other factors, weight appeared to clearly influence reduction of DBP, as the reduction of DBP was more marked the lower the patient's weight. In contrast, the body mass index was poorly correlated with the reduction in blood pressure. The blood pressure reduction, regardless of weight, was more marked when a stricter blood pressure objective was adopted (DBP < or = 80 mmHg). Finally, in the particular case of elderly subjects, treatment with a calcium channel blocker appeared to be very effective in reducing the blood pressure. PMID- 9749165 TI - [Identification and prediction of responders to a therapy. A model and its preliminary application to hypertension]. AB - The effect of a given treatment for a given disease may be estimated from randomized controlled clinical trials, expressed as a single treatment effect averaged over the trial population. However, in recent years there has been an increasing willingness to individualize therapeutic decisions. The method we report here identifies responders by assessing the individual probability of an event, according to the treatment group. We used a treatment-stratified Cox regression model including interaction between treatment and patient's covariates, with common regression coefficients for treated and untreated, except for the special case of a prognostic variable which has an interaction with treatment. Further, we used a discriminate function based on the final model, representing the absolute individual therapeutic effect, to identify the patients to be treated according to a given threshold of clinical efficacy. The model was explored on the INDANA database (which pools individual patient data from clinical trials of anti-hypertensive drug intervention). Data on 36,444 patients, from five randomized controlled trials were included. The results show the relationship between the proportion of avoided events among the avoidable ones, and the proportion of patients treated who were responders, as a function of the threshold of absolute benefit defining responders. The confidence intervals of the absolute therapeutic benefit for each individual were calculated, by using the Monte Carlo simulation method. A comparison of the survival of treated and controlled individuals, in both subgroups of responders and non responders, illustrated the relevance of the model. We conclude that the tools for predicting individual therapeutic benefit do exist. It will be important to assess the reproducibility of these results in other models or in other populations before widespread application. It will be necessary to have a properly computerized environment and to train doctors to use these tools. PMID- 9749166 TI - [Ischemic renal diseases have become the most frequent causes of end stage renal disease in the elderly]. AB - In the 80s it was established that atherosclerotic renal artery stenosis (ARAS) is a leading cause of renal insufficiency and that this condition ranks among the rare etiologies of chronic renal failure amenable to improvement or stabilization particularly in the white. Nephroangiosclerosis (NAS) is an increasing cause of ESRD in the western countries, especially in blacks. Epidemiological data on the vascular nephropathies leading to ESRD are still rare. In this study, we compare annual incidence of ESRD due to ARAS and NAS during two five-year periods: period A = 1982-1986, period B = 1992-1996. The region of the survey comprised 410,664 inhabitants (99.6% of Caucasians), of whom 100,230 were aged over 60 years. Diagnosis of ARAS required arteriography and that of NAS a renal biopsy. Undetermined vascular nephropathy was diagnosed when ESRD patients had had previously no arteriography or no histological examination. Major results were as follow (A vs B, incidence = n/million inhabitants): 1) Increasing incidence of ESRD due to all causes: 76 vs 95 per million, mean age at ESRD 56 vs 62 yrs, percentage of patients over 65 yrs 28 to 59% (p < 0.001). 2) Increasing incidence of ESRD due to vascular nephropathies: 5.5 vs 27.5 per million (p < 0.0001) in general population and 22 vs 110 per million (p < 0.0001) in population aged over 60 years, mean age at ESRD 68 vs 73 yrs. 3) Increasing incidence of different types of ischemic renal diseases leading to ESRD: ARAS 2.5 vs 15 per million (p < 0.0001) in general population and 10 vs 60 per million (p < 0.001) in those aged over 60 yrs, mean age 69 vs 74 yrs, NAS: 1 vs 8 and 4 vs 32 per million (p < 0.001), mean age 67 vs 72 yrs, undetermined VN 0.5 vs 2.5 and 2 vs 10 per million, 65 vs 73 yrs. Our study demonstrates that ischemic renal diseases 1) have become the most frequent causes of ESRD (27% of all patients and 43% of those aged over 6C years) in the Caucasian elderly. 2) are the only cause of increasing incidence of ESRD in this French region. PMID- 9749167 TI - [Genetic study of renal artery fibromuscular dysplasia]. AB - The aim of this study was to conduct a formal pedigree analysis of the involvement of the elastin gene in families. From 140 subjects with renal FMD documented on angiography, family cases with documented renal artery fibromuscular dysplasia (FMD) and to test pedigrees were constructed and familial cases defined by angiographic evidence of FMD in at least one sibling. Familial screening was made either by echodoppler for asymptomatic subjects or by digital intravenous angiography for hypertensive subjects. Linkage analysis at the elastin gene locus was performed in these families with two polymorphic markers: one diallelic RFLP located in exon 16 and one multiallelic CA repeat located in intron 17 of the elastin gene. Fourteen pedigrees (10%) were obtained including nine sibling pairs, four trios and one vertical transmission from a father to his daughter. Most affected subjects were females (84%) but familial cases were more frequently bilateral than sporadic cases (80% vs 49%, p = 0.07). Pedigrees analysis was compatible with an autosomal dominant mode of inheritance and suggested in these families an age and sex-dependent incomplete penetrance model. Linkage analysis resulted in a maximum two-point lod score of 0.06 at theta = 0.20 using the dinucleotide CA repeat. Analysis of the diallelic marker revealed similar frequencies in affected and non affected subjects. This study highlights the role of genetics factors in approximately 10% of FMD cases. The elastin gene does not seem to be involved in the pathogenesis of FMD. PMID- 9749168 TI - [Transplant renal artery stenosis: long term effect of angioplasty on arterial pressure control and renal function]. AB - We assessed the long-term (M +/- SE: 68 +/- 3 months) arterial pressure and renal function of cadaveric kidney transplant recipients with and without significant (> 70% diameter reduction) transplant renal artery stenosis (TRAS) at angiography. Baseline clinical, immunological and outcome data for 26 patients with TRAS (incidence of TRAS: 6.6%) before and following angioplasty and 72 patients without stenosis at angiography were reviewed and analyzed. The 2 groups were similar with respect to recipient sex ratio and age (45 vs 46), duration of transplantation (7 months), cause of renal failure, donor sex and age, HLA antigen mismatches and titers of anti-HLA antibodies, CMV infection and anti-CMV antibodies in donors and recipients. The technical success of angioplasty was 92.3%. Restenosis was documented in 6/26 patients (23.1%). Revascularization resulted in a decrease of arterial pressure and number of antihypertensive medications and a lower serum creatinine compared to baseline values. The long term arterial pressure and serum creatinine levels were similar in patients with and without stenosis. In conclusion, TRAS after revascularization had no detectable influence on the long-term arterial pressure control and renal function within a follow-up period of 68 +/- 3 months. PMID- 9749169 TI - [Renal artery stenosis and chronic renal failure in NIDDM]. AB - The NIDDM patient, willingly with high blood pressure and atheroma, has frequently an abnormal renal function. Must a renal artery stenosis (RAS) be searched as a determining or favorising cause? We have searched RAS by color duplex scan, in 60 consecutive NIDDM patients with altered renal function (creatinine clearance < or = 60 mL/min). Metabolic blood pressure (ABPM), cardiovascular and renal investigations have been realised. The population was composed of 22F/38M with middle age: 70.7 +/- 6.2 yrs, diabetic duration: 11.6 +/ 8 yrs, the plasma creatinine was: 161 +/- 78 mumol/L and clearance: 40 +/- 13 mL/min. Thirty eight had albuminuria, 28 had plasma creatinine > or = 150 mumol/L. All patients had high blood pressure. Significative RAS (> or = 70%) was detected in 15 patients (25%) by color duplex scan and proved with arteriography (n = 10) or angio NMR (n = 5). Twelve (80%) had unilateral stenosis (4 thrombosis), 3 (20%) bilateral stenosis. Renal US lead the diagnosis in 10 patients (66%): unilateral or bilateral hypotrophy. Those 15 patients had these following characteristics: 4F/11M (sex R : 0.36), middle age: 70.8 +/- 7.2 yrs, diabetic duration: 14.3 +/- 7.5 yrs, HbA1c was at 8.4 +/- 2%, 8 (53%) patients require insuline and 5 have retinopathy, plasma creatinine was at 169 +/- 6 mumol/L; 32% of patients with plasma creatinine > or = 150 mumol/L had RAS (n = 9/60%), creatinine clearance was at 38 +/- 12 mL/min (7/47% < or = 30 mL/min), 9 (60%) had macroalbuminuria and 5 (33%) microalbuminuria. All hypertensive patients were treated (mean SBP: 148 +/- 16, mean DBP: 82 +/- 7 mmHg) and had 62 +/- 28% SBP escape and 33 +/- 19% DBP escape. Ten had severe hypertension (at least 3 hypotensive drugs), 12 received CEI; 8 (53%) were smokers; 14 (93%) had one or more macroangiopathies (10/66% coronary heart diseases, 7/46% lower limbs arteritis, 6/40% carotid atheroma); 13 of these macroangiopathies are severe. In conclusion, renal failure (especially evolutive and/or treated with CEI) in NIDDM must call up a RAS (25%) specially in elderly males with a long diabetes duration, severe hypertension and macroangiopathies. This patient profile must lead to a color duplex scan to confirm the diagnosis already suspected by the renal echography. PMID- 9749170 TI - [Modulation by nitric oxide of vasopressin induced renal vasoconstriction varies with perfusate viscosity in the isolated rat kidney]. AB - Previously we reported that AVP is a potent vasoconstrictor in the TYRODE's perfused rat kidney. In vivo however AVP elicited only minor effects on renal blood flow. We hypothetized that differences in shear stress, particularly related to differences in viscosity could be involved. In this study, we investigate the role of perfusate viscosity in the modulation of AVP-induced renal vasoconstriction by NO. Experiments were performed in kidneys isolated from male Sprague-Dawley rats (220 g). Kidneys were perfused at a constant flow of 8 mL/min, in a recirculating system, with TYRODE's solutions supplemented with 6% bovin serum albumin (BSA) or 4.7% Ficoll 400 (Ficoll). The viscosities relative to water were respectively of 1.33 (BSA), 2.32 (Ficoll) and 1.03 (TYRODE). Concentration-response curves to AVP were constructed in the absence or presence of 100 microM N omega-nitro-L-arginine (L-NA), an inhibitor of NO synthase, and compared to those obtained in kidneys perfused with TYRODE's solution. AVP elicited a concentration-dependent renal vasoconstriction, with a progressive shift of the curves to the right and a small decrease in the maximum response when the kidneys were perfused with perfusates of increasing viscosities: logEC50 = -9.9 +/- 0.1 (TYRODE, n = 14), -9.7 +/- 0.1 (BSA, n = 5), -9.0 +/- 0.1 (Ficoll, n = 5) (m +/- e.s.m. Anova, p < 0.001); Emax = 34 +/- 1, 31 +/- 2 and 26 +/- 3 mmHg/mL/min (Anova, p < 0.001). L-NA abolished the differences between kidneys perfused with solutions of different viscosities in logEC50 for vasopressin (10.3 +/- 0.1, 10.4 +/- 0.1 and 10.5 +/- 0.1, n = 5-11, Anova, NS) but did not affect Emax values. In conclusion, present results show that 1) AVP-induced renal vasoconstriction is modulated according to the viscosity of perfusate and 2) NO is involved in this effect. Viscosity, a major determinant of shear stress, should be considered in hemodynamic studies performed on isolated kidneys. PMID- 9749171 TI - [Familial aggregation of blood pressure over 17 years in children and young adults]. AB - Familial aggregation of blood pressure is well known although its causes remain controversial. The aim of the present study is to evaluate the presence of a familial aggregation for blood pressure and body mass index over a 17-year period, in order to evaluate the importance of a primary prevention strategy beginning in familial environment. DESIGN AND METHODS: A longitudinal cohort study was constructed from two cross-sectional surveys 17 years apart: 1,032 individuals, of both sexes, aged 5 to 24 years were seen in the initial study, and their parents whenever possible. Correlation coefficients and stepwise regression analysis were used to analyse the relationship between parents and children. RESULTS: The correlation between parents' and children BP are: systolic BP-0.34 (p < 0.01) and diastolic BP 0.19 (0.05); and for the anthropometric variables are: height-0.29 (0.01); weight-0.41 (p < 0.01); ponderal index -0.21 (p < 0.05); tricipt skinfold-0.21 (p < 0.05). All the coefficients are statistically significative. The variance of children's SBP and DBP explained through a stepwise regression analysis was 47%. The children's weight, skinfold, ponderal index, and parents' SBP and DBP were accepted by the model. CONCLUSION: The relation between BP and obesity variables suggest that a large proportion of familial aggregation for BP may be explained by aggregation for obesity, still after 17 years. PMID- 9749172 TI - [Automobile drivers' licensing and heart rhythm disorders. On what criteria should recommendations be bases?]. PMID- 9749173 TI - [Isolated prolapse of the posterior leaflet of the mitral valve. Results of reconstructive surgery]. AB - Out of 522 patients undergoing mitral valve reconstruction for mitral regurgitation between 1988 and June 1994, the authors studied 159 cases of isolated mitral regurgitation by prolapse of the posterior mitral leaflet. There were 98 men (62%) and 61 women (38%), with an average age of 58.4 +/- 10.4 years. The functional class and ejection fraction were 2.8 +/- 0.11 and 0.66 +/- 0.2 respectively. In 155 patients, surgery consisted in quadrangular resection of the prolapsed tissue, followed in 83 cases by sliding posterior valvuloplasty and in 72 cases by plicature of the annulus. In 4 cases, the prolapse was treated by implantation of artificial chordae tendinae. A Carpentier-Edwards ring was inserted in all cases. There were no hospital deaths. Echocardiography was performed before discharge from hospital and showed satisfactory mitral valve function in 98% of cases: slight systolic anterior motion (SAM) was observed in one case. All patients were followed up for an average of 3.67 +/- 0.10 years. At six years, survival was 93 +/- 7%; moreover, 93 +/- 7% and 97 +/- 3% of patients had no thromboembolic or haemorrhagic complications. Six patients were reoperated, three of them in the first year of follow-up. At six years, 95 +/- 5% of patients were free of reoperation and 81 +/- 11% were free of all complications. The authors conclude that the excellent medium term survival and the low rate of complications are evidence in favour of conservative surgery for treatment of mitral regurgitation due to prolapse of the posterior mitral leaflet. PMID- 9749174 TI - [Long-term follow-up of anatomic heart transplantation. Apropos of 60 patients with a mean follow-up of 36 months]. AB - The aim of this study was to collect different problems seen in long-term evolution of patients who had anatomical cardiac transplantation and to compare with those seen in patients with standard transplantation. During the mean follow up of 36 months, we analysed different data of 60 patients mean aged 51, who underwent anatomical cardiac transplantation. Six patients (10%) died within the 30 days after surgery. No patient needed the use of permanent pacemaker. Echocardiographic examination found normal atrial shape. One month after surgery, echocardiography described 16 tricuspid regurgitations (22.66%) and 8 mitral regurgitations (13.33%), 1 year later, there was respectively 13.33 and 6.66% tricuspid and mitral regurgitation. We had 8 late deaths: 1 sudden death, 2 chronic rejections, 1 pancreatitis and 4 cancers. The survival analysis pointed out 84% at 1 year, 80 at 2 years, 78 at 3 years and 73 at 5 years. Six months after surgery, 80% of patients were treated for high blood pressure; 85% had serum creatinine level equal or superior to 13 mg/L, with mean serum ciclosporin at 130 ng/mL. At the 3rd month, 6 endomyocardial biopsies were equal or superior to grade 2 rejection (International Society for Heart Transplantation). Between the 3rd and 12th month, 3 endomyocardial biopsies were equal or superior to grade 2 rejection, and the same between the 12th and 24th month. The infections rate was 0.8 episode per patient. Long term follow-up of anatomical cardiac transplantation faces the same problems as in standard cardiac transplantation. It is better to perform anatomical cardiac transplantation because of its early postsurgical advantages. Long term care is the same as in standard cardiac transplantation. PMID- 9749175 TI - [The value of transesophageal echocardiography for the diagnosis of pulmonary embolism with acute pulmonary heart disease]. AB - Transoesophageal echocardiography is a method of visualising intracardiac thrombi and could therefore be useful for the diagnosis of pulmonary embolism, but its diagnostic value is unknown. The authors carried out a prospective study with this diagnostic tool in massive pulmonary embolism. The study protocol was to perform transthoracic echocardiography in patients with suspected acute pulmonary embolism and then to perform transoesophageal echocardiography when there were signs of acute cor pulmonale. The results of both echocardiographic investigations were compared with two reference radiological techniques: the spiral CT scan and/or pulmonary angiography. Fifty-six patients underwent transthoracic echocardiography. In the 34 patients with transthoracic echocardiographic signs of acute cor pulmonale, the positive predictive value of the investigation for pulmonary embolism was 91% and the negative predictive value was 54%. Twenty of these 34 patients underwent transoesophageal echocardiography. The sensitivity and specificity for the diagnosis of proximal embolism were 85% and 86% respectively. The limitations of the method were poor visualisation of the left pulmonary artery in which only one thrombus was detected, compared with 6 by spiral CT scan, and the absence of visualisation of lobar arteries. Consequently, the real sensitivity of transoesophageal echocardiography for visualisation of all thrombi in the pulmonary arteries in acute cor pulmonale was only 55%. In acute cor pulmonale, the diagnostic value of transoesophageal echocardiography is poor because the sensitivity for visualisation of intra-pulmonary arterial thrombi is low compared with other radiological techniques. However, in patients with proximal emboli in the right or main pulmonary artery, the diagnosis may be established in a few minutes without the need of other more invasive techniques. Nevertheless, normal transoesophageal echocardiography does not rule out the presence of proximal in the left pulmonary artery or distal emboli in the lobar arteries. PMID- 9749176 TI - [Coronary surgery on the beating heart under extracorporeal circulation in high risk patients. An acceptable compromise?]. AB - Coronary artery surgery with cardioplegia in high risk patients carries a risk of myocardial ischaemia and, without cardiopulmonary bypass, is not always technically feasible. The authors assessed an alternative, surgery on the beating heart with haemodynamic assist by cardiopulmonary bypass in 43 consecutive patients with poor left ventricular function (mean ejection fraction: 0.26), evolving myocardial ischaemia or acute myocardial infarction, old age (mean: 79.5 years) and comorbid conditions. Results were assessed mainly on clinical criteria. In addition, 9 patients had pre- and post-cardiopulmonary bypass measurements of markers of myocardial ischaemia (troponine Ic) and systemic inflammation (interleukines 6 and 10, elastase). In 6 cases, right atrial biopsy was analysed for expression of messenger ribonucleic acid coding for heat shock protein (HSP) 70; the data were compared with those of patients operated under warm blood cardioplegia. There was one cardiac death and one myocardial infarction. Myocardial conservation was confirmed by the minimal increase in troponine Ic levels and the significant increase in HSP 70 in RNA suggesting myocardial adaptation to stress. On the other hand, the minimal concentrations of mediators of inflammation were not significantly changed. In selected high risk patients, coronary revascularisation on the beating heart under cardiopulmonary bypass could be a valuable alternative. It conserves the potentially deleterious effects of cardiopulmonary bypass but peroperative global myocardial ischaemia, an important factor in the aggressivity of cardiac surgery, is eliminated. PMID- 9749177 TI - [Comparison of gas exchange and hemodynamic variables during 2 types of exercise tests. Cycle-ergometry and the ergometric table]. AB - The development of stress echocardiography on an ergometric table has increased the number of stress tests in the decubitus position, whereas most of the information currently available concerns stress tests in the sitting position or on the treadmill. In order to study the influence of this position of stress testing, the authors compared the results obtained in a series of 15 patients without cardiac disease (Group I) and another series of 15 coronary patients (Group II) undergoing the two types of stress testing, in the vertical position on a bicycle ergometer and in the lateral decubitus position on the ergometric table. Effort tolerance on the bicycle ergometer was significantly greater in terms of work load (202 +/- 35 vs 180 +/- 36 watts (p < 0.001) in the controls, and 120 +/- 32 vs 106 +/- 22 watts (p < 0.05) in the coronary group), of duration of effort (19 +/- 3 vs 16 +/- 3 minutes (p < 0.001) in the controls and 10 +/- 3 vs 8 +/- 2 minutes (p < 0.05) in the coronary patients), of heart rate (190 +/- 10 vs 172 +/- 21 beats/min (p < 0.005) in controls and 118 +/- 19 vs 111 +/- 14 beats/min (p < 0.05) in the coronary patients). On the other hand, blood pressure and O2 saturation tended to be greater during exercise in the decubitus position: SBP 200 +/- 23 vs 196 +/- 27 mmHg (NS) in the controls and 158 +/- 21 vs 166 +/- 23 mmHg (NS) in the coronary patients; DBP 97 +/- 10 vs 102 +/- 27 mmHg (NS) in the controls and 85 +/- 6 vs 90 +/- 10 mmHg (NS) in the coronary patients; O2 sat 96.8 +/- 1 vs 97.6 +/- 0.8% (p < 0.05) in the coronary patients. The anaerobic threshold and peak VO2 were much higher during exercise in the sitting position: oxygen consumption at the threshold 14.8 +/- 3.8 vs 12.6 +/- 2.3 ml.kg-1.min-1 (p < 0.01), peak VO2 22.2 +/- 5.9 vs 18.8 +/- 4.7 ml.kg-1.min-1 (p < 0.01) in the coronary patients. The results of this study show that the cardiovascular stimulation obtained in the decubitus position is not identical to that obtained by traditional exercise stress testing, particularly in coronary patients. PMID- 9749178 TI - [Measurement of the left ventricular mass using MRI with automatic determination of the endocardial and epicardial contours]. AB - This study describes a method of automatic border detection of the left ventricular endocardium and epicardium associating three methods of segmentation (increase of region, border detection and adaptive threshold), applicable to the evaluation of ventricular mass and volume by magnetic resonance imaging. Despite slight underestimation, the spin-echo sequence used in 9 small pigs provided a value of left ventricular mass close to that observed ex vivo (r = 0.97, SEE = 6.05 g). Clinical validation using a rapid gradient-echo sequence was undertaken and compared with manual border detection carried out by three independent, trained operators. The study population included healthy subjects and patients with global or segmental left ventricular dysfunction with or without ventricular deformation. The correlations between automatic and manual detection were satisfactory both for calculation of left ventricular mass (r = 0.93, SEE = 13 g) and measurement of surfaces (r = 0.91, SEE = 1.4 cm2). The concordance of the two methods was confirmed by the Bland and Altman test. Cardiac magnetic resonance imaging may provide accurate and reproducible measurements of left ventricular mass within acceptable acquisition and image processing times for routine use. Although the clinical value of such a method is accepted both for establishing the prognosis and guiding management, studies of the cost/efficacy ratio should be undertaken to situate magnetic resonance imaging with respect to other non invasive techniques of investigation of left ventricular function. PMID- 9749179 TI - [Evaluation of the delays in treatment of myocardial infarction. Results of a survey in Alsace]. AB - In order to assess the conditions of access to emergency care of acute myocardial infarction in Alsace, the authors carried out a survey in all hospitals and medical clinics in the region. All subjects admitted for acute myocardial infarction in the region between 3rd December 1995 and 3rd April 1996 were included. The study population comprised 405 persons. The onset of symptoms usually occurred at the patient's home (85% of cases). The first call was made to the general practitioner in 65% of cases. The emergency ambulance transported 40% of patients. The median time to hospital admission was 5 h 15 (average 21 h); the delay was greater in patients over 65 years of age (6 h 42 versus 3 h 51, p < 0.01). This mainly resulted from a delay in calling the doctor by the patients. Thirty nine per cent of patients underwent a myocardial revascularisation procedure (thrombolysis: 27%, direct coronary angioplasty: 12%). Therefore, ten years after a similar study, this survey shows that the delay to hospital admission has not improved and is still too long for effective emergency therapy to be given. In a region where ischaemic heart disease accounts for 10% of all deaths, a multidisciplinary approach is required to elaborate a regional policy for optimising the management of acute myocardial infarction. PMID- 9749180 TI - [Late-occurrence hemolytic anemia after mitral valve bioprosthesis. 2 cases]. AB - The authors report the cases of two patients admitted to hospital for investigation of haemolytic anaemia. Both had undergone, 10 and 12 years previously, mitral valve replacement with a Ionescu-Shiley bioprosthesis. In both cases, in the absence of signs of cardiac failure, Doppler echocardiography showed mitral regurgitation. The association of haemolytic anaemia and dysfunction of the bioprosthesis led to redux valve replacement and correction of the anaemia. Haemolytic anaemia was the presenting sign of bioprosthetic valve dysfunction requiring replacement of the prosthesis. This complication is common with mechanical valve prostheses but much more rare in bioprosthetic valves. PMID- 9749181 TI - [Myocardial infarction due to spontaneous coronary dissection during the postpartum period]. AB - The authors report the case of a 34 year old woman, admitted to hospital because of myocardial infarction two months after delivery of her fifth child. The infarction was caused by spontaneous dissection of the left main coronary and left anterior descending arteries. Twenty-three months later, the patient was well with medical therapy. This case is an example of spontaneous post-partum coronary dissection which is the commonest cause of infarction occurring in that period. The physiopathology of this complication is not fully understood. The prognosis is poor, lethal in two thirds of cases. However, it must be emphasised that coronary dissection may regress spontaneously. Patients were previously referred systematically for surgery, but now it is usually reserved for cases with persistent myocardial ischaemia despite medical therapy. Angioplasty may be preferred despite the risks and may be successful in some cases. PMID- 9749182 TI - [Atrial defibrillators or implantable atrioverters. Initial results]. AB - The atrial defibrillator is a new non-pharmacological treatment of atrial fibrillation (AF) for restoration of sinus rhythm. This device has two programmable modes: automatic or activated by the physician or patient. In the automatic mode, the device delivers a shock synchronous with the R wave to restore sinus rhythm when AF is detected. Two patients with paroxysmal AF resistant to pharmacological therapy were included in a study to assess the efficacy and safety of the atrial defibrillator in the mode activated by the physician. The device implanted in the pectoral region is connected to 3 electrodes, two for atrial defibrillation and sensing positioned in the coronary sinus and right atrium respectively and a sensing and pacing electrode in the right ventricle. The right ventricle is paced if a post-shock pause is detected. It is possible to interrogate the device with a programmer using its Holter function and so determine the number of episodes of AF sensed and treated. The number, intensity and energy of the shocks and the parameters of ventricular stimulation are programmable. In these two patients, the atrial defibrillator effectively reduced prolonged episodes of AF with a follow-up of 12 and 7 months. No pro-arrhythmic effects were observed. Further clinical evaluation is under way to assess this new mode of treatment, including the mode activated by the patient, safety and tolerance of the shocks. In our two patients, the treatment of prolonged episodes of AF was followed by reduction of many short or asymptomatic episodes. PMID- 9749183 TI - [Communication between the left ventricle and the right atrium in infectious endocarditis. Diagnosis using Doppler-echocardiography]. AB - The diagnosis of a communication between the left ventricle and right atrium was made by transthoracic and transoesophageal echocardiography in a 67 year old man with a recurrence of a methicillin-resistant staphylococcus aureus infectious endocarditis complicating aortic valve replacement with a bioprosthesis seven weeks previously. This diagnosis was confirmed at surgery; the left ventricular right atrial communication was closed by suturing its edges and a new aortic valve prosthesis was implanted. Unfortunately, the patient died 4 months later of myocardial dysfunction although the infectious endocarditis seemed to have been sterilised by antibiotic therapy. Doppler echocardiography, especially using the transoesophageal approach is the best diagnostic method for rare complications of infectious endocarditis, usually of the aortic valve, the prognosis of which is improved by early surgery and appropriate antibiotic therapy for the causal organisms. PMID- 9749184 TI - [Mycotic aneurysm in acute bacterial mitral valve endocarditis. Apropos of a case]. AB - A 24 year old man presented with acute endocarditis of the mitral valve. Rupture of a mycotic cerebral aneurysm on the 20th day was successfully treated by interventional catheterisation. Several days later, he underwent mitral valvuloplasty under good conditions. The postoperative period was uncomplicated but emergency surgery was required for a mycotic aneurysm of the superior mesenteric artery. The patient was discharged from hospital without severe neurological sequellae and with a continent mitral valve. PMID- 9749185 TI - [Medical progress is as rapid and important in its management as that in its contents]. PMID- 9749186 TI - [The impact of the data collection method on the medico-economic classification of admissions for myocardial infarction at the public hospitals in Lyon]. AB - The aim of the PMSI (Programme de Medicalisation du Systeme d'Information) is to describe the activity of hospitals for budget allocation. To control the quality of this information, the authors carried out a study comparing the classification in homogenous disease groups (HDG) obtained from the PMSI with that obtained from the epidemiological data base of the PRIMA trial for patients admitted to the Civil Hospitals of Lyon for myocardial infarction between September 1st 1993 and January 31st 1995. Six hundred and fifty standardised hospital summaries were reconstituted form PRIMA data and grouped using the GENRSA 3 software. Five hundred and forty-one of these hospital stays were found in the PMSI data base and grouped. The concordance not due to chance between the two classifications was then assessed by the global kappa coefficient. It was less than the 40% threshold under which concordance not due to chance is considered to be unlikely. The discordances were essentially due to the presence of an associated diagnosis classifying the hospital stay in the HDG corresponding to complicated myocardial infarction. The presence of a classifying associated diagnosis was observed significantly more often in the PRIMA than in the PMSI data base. This results in an underestimation of the hospital activity and could have important repercussions in terms of budget allocation. PMID- 9749187 TI - [Implantation of coronary stents in diabetic patients. Short- and medium-term clinical results]. AB - Coronary balloon angioplasty is associated with a high incidence of restenosis in diabetics and of revascularisation of the culprit lesion and increased long-term mortality. The authors report the short and medium-term results of coronary stenting in diabetics. Between May 1995 and April 1997, 2,182 patients underwent coronary stenting. This population included 272 diabetics : 58 insulin-dependent and 214 non-insulin dependent (oral antidiabetics). Stents were implanted in vessels with mean diameters of 3 mm and over. During the hospital period, the complication rate (mortality, non-fatal myocardial infarction, emergency coronary bypass surgery, subacute thrombosis of the stent) was comparable in non diabetics, insulin-dependent and non-insulin-dependent diabetics (2.55, 0 and 2.0% respectively). One patient (0.5%) died and another (0.5%) had non-fatal myocardial infarction (thrombosis of the stent) in the non-insulin-dependent group. No complications were observed in the insulin-dependent diabetic group. The mean clinical follow-up of 13 months (3-26 months) was respected in 93 and 97% of diabetics. The non-fatal myocardial infarction rate and revascularisation of the culprit lesion were comparable in the insulin and non-insulin-dependent groups (0 versus 0.5% and 8.2 versus 10.5% respectively) but global mortality was higher in the insulin-dependent diabetics (9.3 versus 2.4%). PMID- 9749188 TI - [Mitral valvuloplasty for asymptomatic or pauci-symptomatic mitral insufficiency]. AB - The aim of this study was to assess the results of mitral valvuloplasty for chronic asymptomatic or paucisymptomatic mitral regurgitation. Of 584 patients operated for chronic mitral regurgitation between January 1989 and December 1994, 175 were in NYHA Classes I and II and made up the study population. All had chronic grade 3 or 4/4 mitral regurgitation suitable for mitral valvuloplasty. The average follow-up was 34.3 months. Mitral valvuloplasty was performed in 174 patients, the other patient requiring mitral valve replacement. Three patients died (1.7%) and the actuarial 5 year survival was 98.2 +/- 1.0%. The probability of absence of reoperation and absence of thrombo-embolic complications at 5 years were 97 +/- 0.8% and 96.3 +/- 1.7% respectively. The residual regurgitation at Doppler echocardiography was minimal or absent in 94% of patients at the last follow-up control. The mean end-systolic and end-diastolic left ventricular dimensions decreased from 40.0 +/- 6.8 mm and 64.8 +/- 7.0 mm before surgery to 34.6 +/- 6.7 mm (p < 0.001) and 52.7 +/- 7.4 mm (p < 0.001) at the last control. The authors conclude that conservative mitral valve surgery for NYHA Classes I and II patients with chronic mitral regurgitation is feasible with a low risk and is associated with a significant reduction in ventricular volumes and stability of valvular continence at medium-term. When performed by teams trained in techniques of mitral valvuloplasty, these results suggest that surgery should be performed early. PMID- 9749189 TI - [Blood flow in the isovolumetric relaxation phase in heart transplant patients. Its use in the noninvasive diagnosis of acute rejection]. AB - The aim of this study was to assess a Doppler-echocardiographic parameter which has not been previously reported for the diagnosis of acute cardiac rejection. The parameter was left ventricular isovolumic relaxation blood flow. Eighty patients who had undergone orthoptic cardiac transplantation were followed up regularly with echocardiography for a period of 2 years. In all, 495 echocardiographic studies were performed and the results compared with those of endomyocardial biopsy performed on the same day (11.4 echocardiographic studies per patient). In the absence of cardiac rejection, isovolumic relaxation Doppler signal was recorded in all patients (364/387 echo studies). This was a positive signal directed towards the apex detected by continuous mode Doppler in the apical position, arising along the interventricular septum in the mid part of the left ventricle (82% of cases) or from the basal region of the septum (18% of cases) and lasting throughout the phase of isovolumic relaxation. The maximal velocity was 0.53 +/- 0.08 m/s (range 0.32 to 0.73 m/s) : the velocity-time integral was 34 +/- 33 cm. This signal was associated with medioventricular endosystolic acceleration of blood flow in 75% of cases. The incidence of the isovolumic relaxation flow signal decreased in cardiac rejection with no significant changes in the other usual Doppler-echocardiographic parameters except for a significant decrease in the ejection fraction in the group with severe rejection. In the group with mild rejection (n = 89) an isovolumic relaxation flow signal was only observed in 52 cases (including 29 in whom immunosuppressive treatment was not increased). In patients with moderate rejection (n = 12) there were only 5 cases in which a isovolumic relaxation flow signal was recorded, and in the group with severe rejection (n = 7), the signal could only be recorded in 1 case. The authors conclude that the absence of an isovolumic relaxation blood flow signal in a cardiac transplant patient is a reliable sign of cardiac rejection with an excellent specificity (94%). The absence of this signal is a sensitive indicator of severe rejection (86%) but less so for moderate (58%) or mild rejection (42%). PMID- 9749190 TI - [Dual-chamber implantable automatic defibrillators. Experiences apropos of 16 cases]. AB - In view of the large number of inappropriate shocks observed in patients with implanted defibrillators, improved detection of ventricular arrhythmias has become a major objective. The addition of an atrial catheter has been proposed to improve discrimination between ventricular and non-ventricular arrhythmias. Besides this function, the additional catheter could be used for DDD pacing without risk of interaction between the pacemaker and defibrillator. The authors report their initial experience in 16 patients implanted with a DDD pacemaker. The indication was resuscitated sudden death (N = 5) or ventricular tachycardia (N = 11). The choice of a DDD defibrillator was justified by a bradycardia (N = 9), haemodynamic factors (N = 4) or supraventricular tachycardia (N = 3). The devices used were the Defender 9001 (ELA Medical SA, France, N = 3), the Ventak AV 1810 and the Ventak AV II DR 1821 (Guidant/CPI, Inc. USA, N = 11 and N = 2 respectively). There were three immediate complications. After 2 to 29 months' follow-up, 5 patients had received appropriate treatment by their devices. Five patients had inappropriate shocks : one patient received a shock triggered by electrical interference, two others had no active sensing algorithme when the shocks were delivered, and the other two had an activated algorithme with 1/1 conduction of a supraventricular arrhythmia. No recurrences were recorded after reprogramming the device. DDD or VDD pacing was permanent in 9 patients and intermittent in 3 others. Seven patients had dilated cardiomyopathy and severe cardiac failure and were clinically improved by dual chamber pacing. In many patients, candidates for a defibrillator, this new generation of devices has improved specificity of arrhythmia detection and cardiac pacing without risk of interaction. The authors propose a classification of the indications for a DDD defibrillator. PMID- 9749191 TI - [Cardiac abscess in infectious endocarditis. A multicenter study apropos of 233 cases. The Working Group on Valvulopathy of the French Society of Cardiology]. AB - The aim of this retrospective multicenter study was to determine present characteristics of infectious endocarditis complicated by abscess and to identifying predictive factors of mortality. The files of 233 patients with infectious endocarditis complicated by perivalvular abscesses between January 1989 and December 1993 were analysed. Two hundred and thirteen patients underwent medico-surgical treatment (175 aortic and 38 mitral abscesses) and 20 patients underwent medical treatment alone (17 aortic and 3 mitral abscesses). The abscess was observed on native valves in 156 cases and valve prostheses in 77 cases. The causative organism was identified in 69% of cases : the commonest organism was the staphylococcus. The diagnostic sensitivity of transthoracic and transoesophageal echocardiography was 36 and 80% respectively. The operative mortality at one month was 16%. Patients over 65 years of age, staphylococcal infection, renal failure and fistulisation of the abscess, were identified as independent predictive factors of mortality at one month. The survival rate three months after surgery was 75 +/- 10% and 59 +/- 11% at 27 months. An age over 65, staphylococcal infection, uncontrolled infection, circumferential abscess and fistulisation were independent predictive factors of global mortality (the first month and after). The mortality rate in unoperated patients was 40%: cardiac failure and fistulisation of the abscess detected by echocardiography were predictive factors of mortality on univariate analysis. PMID- 9749193 TI - [3-dimensional reconstruction technic using power Doppler. In vitro use on normal and stenosed pulmonary artery bifurcations]. AB - Power Doppler imaging is an ultrasonic technique representing circulating red cells as a colour signal independent of their direction and angle of analysis. This noninvasive technique provides a morphological representation of blood flow within a blood vessel. This in vitro study in pulsed flow enabled three dimensional reconstruction of blood flow in the bifurcation of the pulmonary artery with automatic shifting of the ultrasound probe and an integral digital transfer of the echographic data. The software for treating the images and for volumic reconstruction of the flow is explained. This technique provides a quantitative method of assessing residual intrastenotic flow with a simple computer system. PMID- 9749192 TI - [Infections secondary to implantation of cardiac pacemakers]. AB - Infectious complications of pacemaker implantation are not common but may be particularly severe. Localised wound infections at the site of implantation have been reported in 0.5% of cases in the most recent series with an average of about 2%. The incidence of septicaemia and infectious endocarditis is lower, about 0.5% of cases. The operator's experience, the duration of the procedure and repeat procedures are considered to be predisposing factors. The main cause of these infections is though to be local contamination during the implantation. The commonest causal organism is the staphylococcus (75 to 92%), the staphylococcus aureus being the cause of acute infections whereas the staphylococcus epidermis is associated with cases of secondary infection. The usual clinical presentation is infection at the site of the pacemaker but other forms such as abscess, endocarditis, rejection of the implanted material, septic emboli and septic phlebitis have been described. The diagnosis is confirmed by local and systemic biological investigations and by echocardiography (especially transoesophageal echocardiography) in cases of right heart endocarditis. There are two axes of treatment: bactericidal double antibiotherapy and surgical ablation of the infected material either percutaneously or by cardiotomy. Though controversial, and unsupported by scientific evidence, the role of systematic, preoperative, prophylactic antibiotic therapy in the prevention of these complications seems to be increasing. PMID- 9749194 TI - [False aneurysm of the left ventricle. A sometimes late finding]. AB - Left ventricular pseudo-aneurysm is a rare complication of myocardial infarction, usually of the inferior wall. It is generally a sudden event due to rupture of the heart which is contained by the pericardium. The outcome is usually rapidly fatal by secondary rupture or adiastole. The authors report a case of pseudo aneurysm of the left ventricle measuring 3.5 cm in diameter observed following a small inferior wall myocardial infraction in a diabetic patient with a history of inferior wall myocardial infarction 38 years previously. This case is interesting because of the silent character of the pseudo-aneurysm, very probably complicating the previous infarct. PMID- 9749195 TI - [Cardiogenic shock presenting a cardiac sarcoidosis. Apropos of a case]. AB - The authors report a case of cardiac sarcoidosis in a 38 year old patient presenting initially with cardiogenic shock. The diagnosis was made by myocardial biopsy. The patient underwent cardiac transplantation for terminal, refractory cardiac failure but postoperative complications led to the death of the patient a few weeks later. This rare observation should be noted because the causal disease may benefit from specific therapy. PMID- 9749196 TI - [Clinical expression of papillary fibroelastoma. Apropos of a case]. AB - The authors review the literature of the clinical features of papillary fibroelastomas in the light of a new case. These benign tumours of the endocardium may be distinguished from Lambl's vegetations by their site and size. Some workers suggest that they correspond to giant Lambl's vegetation and could be a form of "aging" of the valvular endocardium. Nevertheless, Lambl's vegetations are always present after 10 years of age but the papillary fibroelastoma is rarely detected by echocardiography and there have been few case reports. They are essentially cardiac valve tumours (73% of valvular tumours) and may give rise to serious clinical symptoms, sudden death by migration or coronary obstruction, systemic embolism, especially from left heart lesions. However, they can be situated at any point of the endocardium. The diagnosis of a valvular or an endocardial tumour is based on echocardiography which, though not always accurate, gives a better aetiological diagnosis. In cases of symptomatic tumours, surgery (usually simple ablation) is indicated with a low operative risk and cure of symptoms. Tumours discovered by chance pose very difficult problems of management and may lead to diagnostic or preventive surgery. PMID- 9749197 TI - [Opening ceremonies of the European session of the French Society of Cardiology (15 January 1998)]. PMID- 9749199 TI - [Financial report fiscal 1997]. PMID- 9749200 TI - [Address of the outgoing President]. PMID- 9749201 TI - [Address of the incoming President]. PMID- 9749202 TI - [Pediatric cardiology is a form of preventive medicine]. PMID- 9749203 TI - [Anatomic evaluation of ostium secundum atrial septal defects by tridimensional echocardiography]. AB - The decision to close an ostium secundum atrial septal defect by interventional catheterisation implies knowing its size, form and the relationship of its borders to neighbouring structures as accurately as possible. Three-dimensional echocardiography provides unique views of the interatrial septum and the authors set out to assess its performance. Ten patients, aged 8 to 20 years, included in a multicenter European clinical trial of closure of atrial septal defects with the CardioSEAL prostheses, were examined by transoesophageal echocardiography with three-dimensional reconstruction of the interatrial septum viewed from the left or right atrium. The septal defect had a very variable morphology, round, oval raquet-shaped and occasionally multiple. The surface area of these defects varied by about 70% during the cardiac cycle, maximal during ventricular systole and minimal during atrial systole. The maximal diameter measured by two dimensional transoesophageal echocardiography underestimated that measured by three-dimensional echocardiography by about 30%. Two patients had a juxta-aortic caudal border or a juxta-superior vena caval cephalic border making the defect unsuitable for catheter insertion of a CardioSEAL occluder. On the other hand, another patient had an adequate juxta-aortic border although it seemed too narrow with conventional imaging techniques. The authors conclude that three-dimensional reconstruction of transoesophageal echocardiography is the best method of selecting candidates for closure of ostium septum atrial septal defect by intervantional catheterisation. PMID- 9749204 TI - [Short- and medium-term results of cavo-pulmonary anastomosis]. AB - The authors report the medium term results of total cavo-pulmonary connections in 50 patients treated between 1988 and 1997. POPULATION: the cardiac lesions were single ventricle (n = 27), tricuspid atresia (n = 11), mitral atresia (n = 2), pulmonary atresia with intact septum (n = 2) or more complex lesions (n = 8). The mean age at surgery was 75 +/- 6 months. There were no operative deaths but two early reoperations were required. After surgery, oxygen saturation increased from 76 to 91% (p = 0.001). Early complications included: 11 mild cardiac failures, 19 pericordial or pleural effusions and 6 supraventricular arrhythmias. At medium term (average follow-up of 59 months), there were no fatalities and all were in NYHA Classes I and II. Cyanosis was observed in 12 patients: right-to-left atrial shunt (n = 7), 2 of which were closed percutaneously, shunt from the azygos system to the suprahepatic veins in 5 cases with Kawashima, 2 of which were reoperated, pulmonary arterio-venous fistula (n = 3). Six patients had arrhythmias and 3 had regressive thromboses. At the end of follow-up, 8 patients remained cyanotic, 4 had antiarrhythmic therapy and 2 had mild ventricular dysfunction. In conclusion, the results of total cavo-pulmonary connections are satisfactory at short and medium term and may be proposed for patients with complex univentricular cardiac malformations. PMID- 9749205 TI - [Clinical, hemodynamic and angiographic results of total cavo-pulmonary connection]. AB - The aim of the study was to assess the short and medium term results of total cavo-pulmonary connection based on analysis of the functional status, the cavo pulmonary circulation and the surgical techniques, and the hepatic consequences. Fifteen patients with congenital defects beyond repair were treated by total cavo pulmonary connection at Tours between March 1st 1992 and July 30th 1996. There were 12 children (mean age: 6.3 years) and 3 adults aged 25 to 28. Results were assessed by clinical examination, hepatic function tests and cardiovascular investigations including right heart catheterisation with angiography in 14 patients. There were no fatalities. Seven patients were in functional Class I and 8 in Class II at medium term (average follow-up of 33 months). Hepatic function was mildly abnormal in all patients with an increase in serum bilirubin and gamma GT, and a decrease in the coagulation factors. The mean pressures in the atrial channel were 12 mmHg (9-16 mmHg), in the superior vena 13.2 mmHg (10-18 mmHg), in the right pulmonary artery 9.5 mmHg (7-15 mmHg) and 11.6 mmHg (8-16 mmHg) in the left pulmonary artery. Significant residual stenosis of a pulmonary branch was observed in 2 cases. The cavo-pulmonary anastomoses were out of line, one from the other, in all cases. The atrial channel was tubular in 9 cases and dilated with slight stagnation of the contrast medium in its inferior region in 5 cases. Total cavo-pulmonary connection transformed the clinical status of these patients but was associated with minor abnormalities of liver function. The quality of the cavo-pulmonary circulation and the surgical anastomoses was estimated to be satisfactory in the majority of cases. PMID- 9749206 TI - [Can partial cavo-pulmonary connection be considered an alternative to the Fontan procedure?]. AB - The disappointing long-term results of the Fontan procedure led the authors to assess substitution with partial cavo-pulmonary connections for definitive palliative treatment of single ventricle malformations. One hundred and fifteen patients with a mean age of 4.3 +/- 4.5 years (1 month-22 years) were treated by termino-lateral anastomosis between the superior vena cava and corresponding pulmonary artery, either of necessity because of a contraindication to total cavo pulmonary connections (31 cases) or electively (84 cases). Another source of pulmonary flow was preserved or added in 76% of children, the operative mortality was 4% and the secondary mortality 3.5%. Significant complications were observed in 15% of cases with a secondary morbidity of 13%. Reoperation was required in 18 cases (16%). In fact, the death rate was higher in indications of necessity (19 versus 3.6% in elective procedures). Similarly, the number of serious complications (veno-venous or pulmonary arterio-venous fistulae, ventricular dysfunction) was higher in this group than in patients undergoing an elective procedure (23 versus 2.4%). After 4.8 +/- 3.2 years' follow-up, 73% of children treated of necessity and 95% of children treated electively were well despite mild cyanosis (average saturation of 86 +/- 6%). On condition that these results are confirmed at long-term as present follow-up is relatively short, this strategy would seem to be justified, providing mild cyanosis with little functional impairment is accepted, so avoiding the serious complications of Fontan-like circulations. PMID- 9749207 TI - [Cavo-pulmonary anastomosis associated with intracardiac repair of Ebstein anomaly. Value in high-risk patients]. AB - The surgical prognosis of Ebstein's anomaly depends on the quality of tricuspid valve repair and right ventricular function. In patients with right ventricular failure, a decrease in afterload was attained by a cavo-bipulmonary anastomosis associated with the intraventricular repair. Fifty-nine out of 111 patients operated for Ebstein's anomaly were considered to be at high risk and were selected for this study. The inclusion criteria were one or more of the following factors: massive tricuspid regurgitation, extensive atrialisation of the right ventricle, poor right ventricular function, chronic atrial fibrillation. This population was divided into two groups with the same preoperative features: Group I (45 operated patients: tricuspid valvuloplasty with longitudinal plicature of the right ventricule: Group II (14 operated patients): same intracardiac repair as Group I and associated cavo-bipulmonary anastomosis. The operative mortality was 24% (11/45) in Group I and 7% (1/14) in Group II (p < 0.05). The 5 year actuarial was 68.6% in Group I and 61.8% in Group II (NS). The reoperation rate was 11% (5/45) in Group I and 0% in Group II. In Group II, the persistence of significant tricuspid regurgitation was better tolerated and the frequence of reoperation was decreased with respect to Group I. The authors conclude that high risk patients with Ebstein's anomaly have a lower operative mortality and improved functional tolerance when there is persistent tricuspid regurgitation after cavo-bipulmonary anastomosis. PMID- 9749208 TI - [Aortocoronary bypass in children. Apropos of 6 cases]. AB - The improvement in diagnostic techniques for myocardial ischaemia and in imaging of coronary anomalies in children has increased the number of coronary revascularisation procedures. Moreover, the indications are more diverse with the emergence of acquired coronary disease which was unknown until recently, related to operations comprising manipulation of the coronary arteries. The authors report their experience of 7 internal mammary artery bypass grafts in 6 children aged 3 days to 11.5 years (average 2 years): 3 left main coronary stenosis after arterial switch for transposition of the great arteries, 1 peroperative lesion of an intramural left coronary in a case of simple transposition of the great arteries, 1 congenital atresia of the right ostium associated with Tetralogy of Fallot, and 1 right coronary occlusion with giant aneurysms of the left coronary in a case of Kawasaki's disease. There were no operative fatalities. All the coronary grafts were patent at early postoperative control. These results indicate clearly that the method is very feasible from a very early age. Long term patency, the growth of the anastomosis and of the distal vessels are questions which await a reply in the future. PMID- 9749209 TI - [Does NMR provide information complementary to cardiac catheterization in aortic coarctation?]. AB - The results of magnetic resonance imaging (MRI) in the investigation of coarctation of the aorta were assessed and compared with those of cardiac catheterisation. This was a retrospective study of a series of 24 patients aged 14 +/- 4 years with a coarctation treated and documented by MRI. The investigation was performed with a high field 1.5 tesla (Vision, Siemens) system. Twenty-one children underwent comparative retrograde cardiac catheterisation with angiography and measurement of the peak-to-peak pressure gradient across the isthmus. No significant difference in the measurements of the aorta in MRI spin echo, gradient echo and retrograde aortic angiography were observed. On the other hand, there was a slight correlation between the degree of stenosis measured by MRI and the peak-to-peak haemodynamic gradient (r = 0.40). Seven patients had a loss of signal at the level of the aortic isthmus on MRI angiography which correlated with the haemodynamic gradient (p = 0.04). The authors conclude that MRI is a reliable non-invasive technique of investigating coarctations of the aorta. It gives accurate morphological data concerning the stenosis and blood flow. MRI should be part of the investigations of coarctation of the aorta, especially in poor indication to be able to correct it or consider the results of angioplasty or surgical correction. PMID- 9749210 TI - [Mid-term results of treatment of aortic coarctation in neonates]. AB - Between 1990 and 1997, 122 neonates aged 8.7 +/- 7.5 days, 75 boys (61.4%), were referred for coarctation of the aorta which was isolated (54 cases) or associated with one (52 cases) or more (20 cases) ventricular septal defects. Hypoplasia of the aortic arch, diagnosed in 52 cases, was more common in children with ventricular septal defects (p < 0.05). The diagnosis was later in isolated coarctation (10.6 +/- 6.8 days) than in cases with shunts (7.8 +/- 7.7 days) and/or hypoplasia of the aortic arch (5.1 +/- 4.3 days). One hundred and nineteen patients were operated, including 112 of left thoracotomy (24 had pulmonary artery banding in addition) at the age of 1.1 +/- 2.7 months, and 7 by sternotomy of first intention for aortic repair and closure of ventricular septal defect. After thoracotomy, closure of the ventricular septal defect was undertaken at 11.3 +/- 10.8 months in children who had undergone previous pulmonary banding and at 3.5 +/- 2.4 months in the absence of banding. Early mortality after aortic repair was 2.5% and late mortality 9.5%, higher in cases of large ventricular septal defects and hypoplasia of the aortic arch (p < 0.001). Follow-up varied from 55 days to 7.8 years (3.99 +/- 2.24 years). Global survival was 97.5% at 1 month and 98.2% at 8 years. In coarctation with ventricular septal defect survival was 95.6% at 1 month and 74.7% at 8 years with a worse prognosis in cases with large single interventricular shunts. Restenosis was observed in 28.5% of cases, 2.25 +/- 3.8 months after aortic surgery (88.5% of cases before the 6th month) and was generally treated by percutaneous aortic angioplasty (10 cases performed 13.5 +/- 12 months after surgery). In all, two factors seemed to increase the risk of death (hypoplasia of the aortic arch and large ventricular septal defects) and restenosis was observed in 1 out of 4 cases, usually before the 6th postoperative month. PMID- 9749211 TI - [Study of the cardiorespiratory response and chronotropic function after correction of tetralogy of Fallot. Important role of residual pulmonary regurgitation]. AB - Effort tolerance is reduced after correction of Tetralogy of Fallot. This prospective study investigated the cardiorespiratory response and the chronotropic function (mean follow-up 11.1 years) of 70 patients (43 boys and 27 girls) with an average age of 14.9 +/- 7.2 years (group 1) compared with 65 normal, sedentary subjects paired red for age and gender (group 2). All underwent exercise testing (Bruce protocol) with measurement of respiratory gases. Quantification of pulmonary regurgitation was performed by Doppler echocardiography. The chronotropic response to exercise was assessed by calculating the mean of slopes established by chronotropic metabolic relationship of Wilkoff. The cardiorespiratory response to exercise was abnormal in group 1: the duration of exercise (11.3 vs 13.6 min; p = 0.005), peak VO2 (35.5 vs 46 ml/min/kg; p < 0.001) and anareobic threshold (8.3 vs 9.2 min; p = 0.001) were decreased. Maximal heart rate (172 vs 190bpm; p < 0.001) and the mean of the metabolic-chronotropic slopes (0.68 vs 0.83; p < 0.001) were decreased in the patient group, showing abnormal chronotropic response to exercise. The latter seemed to be related to the severity of pulmonary regurgitation. The duration of exercise (10.6 vs 11.5 min; p = 0.001), peak VO2 (33 vs 37 ml/min/kg; p < 0.001), maximal heart rate (161 vs 177 bpm; p = 0.002) and the mean of the slopes of the metabolic-chronotropic relationship (0.59 vs 0.72; p < 0.001) were decreased in patients with moderate to severe pulmonary regurgitation. This study shows that significant pulmonary regurgitation is responsible for a poor cardiorespiratory response to exercise and for an abnormal chronotropic response which seems to be multifactorial but probably related to an adaptation favouring left ventricular filling during exercise. PMID- 9749212 TI - [5-year results of arterial correction in transposition of great vessels]. AB - A prospective study was performed on in-hospital patients between June 1985 and July 1992 to assess the 5 year results of surgical detransposition of the great arteries. Clinical examination, electrocardiography, echocardiography, right and left heart catheterisation with selective coronary angiography, isotopic right and left ventricular ejection fractions at rest and with infusion of dobutamine and SestaMibi myocardial perfusion scintigraphy at rest and with dipyridamole, were performed during the 5th year after surgery. Twenty-six children underwent this protocol: eight others did not come for examination because they had moved from the region, one of whom had suffered regressive postoperative myocardial infarction. All patients were asymptomatic and had only minor electrocardiographic changes. Stenosis of the pulmonary tract was observed in 38.5% but only one case of stenosis at the origin of the right pulmonary artery required percutaneous angioplasty, which was successful. Pulmonary regurgitation was a common echocardiographic finding (65.4% of cases) but rarely severe (1/26: 3.9%). Aortic regurgitation was also observed commonly (53.8%), nearly always mild, grade I (13/14 cases). No significant stenosis of the aortic anastomosis was observed. The right and left ventricular ejection fractions were normal at rest except in one case and all values improved with dobutamine. Myocardial scintigraphy did not show any perfusion defect and there was no stenosis or occlusion of the coronary arteries at coronary angiography. The authors conclude that the results of arterial detransposition at 5 years are satisfactory in this series, with no cases of major obstructive lesions, major ventriculo-arterial regurgitation, ventricular dysfunction or coronary lesions. However, longer term trials with larger numbers of patients are required to determine the real incidence of coronary lesions and the long-term outcome of the pulmonary valve in the systemic position. PMID- 9749213 TI - [Left intra-atrial membrane without pulmonary vein obstruction: benign condition of progressive evolution? Apropos of 7 cases]. AB - Left triatrial heart is defined as division of the left atrium into two chambers, proximal and distal, by a variably perforated membrane. The data of appearance of symptoms, often in early childhood, is related to the degree of obstruction and the presence or not of an inter-atrial shunt. Widescale usage of echocardiography, the investigation of choice for this diagnosis, has led to the detection of this abnormality in older patients, sometimes asymptomatic, without pulmonary hypertension. Three adults were referred for transthoracic and transoesophageal echocardiography to investigate systemic embolic disease (2 cerebral, 1 mesenteric). Two other adults underwent the same investigations for diagnosis of the aetiology of atrial fibrillation with mitral regurgitation. Two cases were asymptomatic children, one with a clinically benign murmur and the other with ventricular extrasystoles with no malignant features. In these seven cases, transthoracic (n = 5) and/or transoesophageal (n = 7) echocardiography demonstrated a left atrial membrane corresponding to the classical description of cor triatrium. The Doppler study showed no obstruction in 6 cases and minimal obstruction in 1 case. In our series, as in similar cases reported in the literature, the diagnosis of a left atrial membrane did not lead to surgery. Although we do not know the long-term outcome of this abnormality in asymptomatic children, the observations of complications in the adult suggest a potential of evolution which poses the question of optimal management. PMID- 9749214 TI - [Neonatal ventricular tachycardia]. AB - Ventricular tachycardia is rare and poorly understood in the neonate. The authors undertook a retrospective study in 2 foetus and 8 neonates aged 1 to 20 days at the time of diagnosis. The tachycardia was permanent in 2 cases, observed in runs of variable variation in the other 8, incessant in 7 of these cases. Only two cases were symptomatic: cardiac failure with shock 16 hours after birth and hydramnios at 16 weeks gestation. The electrocardiographic criteria of ventricular tachycardia (wide QRS complexes of different morphology to the sinus QRS complexes, atrioventricular dissociation) were fulfilled in all patients. The arrhythmia was monomorphic 9 times out of 10 with a fixed (3 cases) or variable (7 cases) rate which was always > 150/min. Intravenous magnesium sulphate in the severe and permanent forms, oral betablockers in forms triggered by acceleration of the sinus rhythm, oral amiodarone alone or associated in one case with propranolol were prescribed but three neonates were not treated, either from the outset or after inefficacy of amiodarone: nine of the patients were cured and are treatment-free 12 to 24 months later: the other patient has a slow, well tolerated ventricular tachycardia. No aetiology was detected in 9 cases; the other had a metabolic disease of B-oxidation of long chain fatty acids. The authors conclude that isolated, idiopathic ventricular tachycardia of the neonate usually carry a good prognosis which is not dependent on the tachycardia of the permanence of the arrhythmia. Simple treatment (betablocker or amiodarone) is usually associate with restoration of sinus rhythm and definitive cure during the first year of life. PMID- 9749215 TI - [Result and outcome of prenatal detection of congenital cardiopathies at the Cote d'Or over a period of 9 years]. AB - Antenatal echocardiographic diagnosis of congenital heart disease in 699 women living in the department of the Cote d'Or between 1988 and 1996 revealed 39 cardiac abnormalities which were later confirmed by anatomical examination or echocardiography. During the same period, 157 cardiac malformations which could have been detected in the antenatal period were diagnosed after birth. The detection rate of congenital heart disease was 19.8% over the whole study period, and 33.3% in the last three years. The malformations which were most easily diagnosed were hypoplastic left ventricle, single ventricle, atriventricular canal and severe obstruction of the ventricular outflow tracts. Twenty-one abortions (53.8%) were undertaken because of the severity of the cardiac lesion or of a genetic abnormality. Seventeen children were live born (43.5%) but, globally, they had severe malformations and the mortality rate was 84.5% whereas in the children diagnosed post-partum, the lesions were generally less severe and the mortality was 19.7%. The detection rate of congenital heart disease depends on the screening of first intention. Thus, 90% of the lesions diagnosed in this series were referred because of a foetal heart abnormality and, in this indication, echographic expertise was by far the most accurate with 33% of confirmation. As other workers have reported, teaching programmes and regional collaboration between physicians implicated in antenatal care are essential. PMID- 9749216 TI - [Aneurysm of the vein of Galen and cardiac insufficiency. Current therapeutic approach]. AB - Aneurysm of the ampulla of Galien is a rare but serious cause of cardiac failure in the neonate or child. The diagnosis is suspected on finding a continuous murmur on auscultation of the skull. It is an intracranial arterio-venous malformation which is sometimes responsible for a very important shunt between the arterial (carotid or vertebral) and venous systems. The vital and neurological prognosis of these children is classically very poor. The authors report the cases of the last three consecutive children aneurysms of the ampulla of Galien followed up at Grenoble Hospital in the last five years. The clinical presentations were very different, corresponding to the usual three forms described in the literature: a reputedly very severe form with cardiac failure at the 3rd day of life requiring multiple percutaneous embolisations; a neonatal form revealed by an isolated macrocranium; and a form diagnosed late (4.5 years) in the investigation of effort dyspnoea which regressed after two sessions of embolisation. All three children are alive and have normal psychomotor development for their age. A multidisciplinary approach involving neonatal physicians, paediatric cardiologists and neurologists, with complementary non invasive investigations such as transfontanellar ultrasonography, electroencephalography, MRI and echocardiography are necessary to optimise the management and limit the neurological sequellae in children with this type of malformation. Cerebral arteriography and percutaneous embolisation under general anaesthesia by a trained team gave very good medium-term results in two of these cases, lightening the usual pessimism surrounding this condition. PMID- 9749217 TI - [Idiopathic dilatation of the right atrium simulating Ebstein's anomaly. Apropos of a case diagnosed in utero]. AB - In October 1992, severe dilatation of the right atrium was detected in a 35 week foetus. The septal leaflet of the tricuspid valve seemed to be displaced distally causing massive tricuspid regurgitation. The diagnosis of Ebstein's anomaly was made and confirmed after birth. Refractory right heart failure occurred at the age of 10 months and the little girl was operated. At surgery, the right atrium was very dilated; the tricuspid valve was normally positioned and was normally constituted. The right atrial wall, partially resected, contained few muscular fibres and showed patchy fibroelastosis. Four years after surgery, the child remains asymptomatic. Dilatation of the right atrium and the tricuspid annulus caused pseudo-displacement of the septal leaflet of the tricuspid valve. This, combined with the importance of tricuspid regurgitation, led to the erroneous diagnosis of Ebstein's anomaly. It is important to differentiate idiopathic right atrial dilatation from Ebstein's anomaly because surgery is much more difficult in the latter case. PMID- 9749218 TI - [Double outlet right ventricle with sub-aortic obstruction caused by hypertrophy of the parietal band. Echographic image apropos of a case]. AB - The authors report the case of an 18 month old infant with double-outlet right ventricle and subaortic ventricular septal defect with severe subaortic obstruction. Echocardiography showed the subaortic obstruction to be due to severe hypertrophy of the parietal band which grew upwards to the aortic annulus. A peak instantaneous Doppler gradient of 72 mmHg was recorded between the right ventricle and the aorta. The hypertrophy of the band was secondary to pulmonary artery banding performed in the neonatal period at the same time as cure of an isthmic coarctation of the aorta. Surgical exploration confirmed the echocardiographic findings a circular subaortic muscular stenosis related to hypertrophy of the parietal band. Resection of this band liberated the left ventricular outflow tract and made possible a tunnelisation procedure between the left ventricle and the aorta. Postoperative Doppler echocardiography showed the absence of subaortic stenosis. PMID- 9749219 TI - [Fetal supraventricular tachycardia associated with anasarca: poor prognosis despite treatment. Apropos of two cases]. AB - Two cases of foetal supraventricular tachycardia with hydrops with fatal outcomes illustrate the poor general prognosis of this condition. The absence of therapeutic consensus, of large series in the existing literature, does not prevent logical and reasonable management based on rhythmological, pharmacological and prognostic criteria. A combined approach associating antiarrhythmic therapy by the transplacental and intrafunicular approaches seems acceptable now that funicular puncture can be undertaken easily, and certain antiarrhythmic molecules suggest encouraging results. It is important to try to assess the haemodynamic tolerance by foetal Doppler echocardiography because the foetal prognosis depends on ischaemic cerebral lesions induced by anoxia, difficult to diagnose in utero: when observed, aggressive and occasionally dangerous therapies to foetus and mother may be justified in these extreme situations of foetoplacental hydrops. PMID- 9749220 TI - [Recurrent false infundibulo-pulmonary aneurysm after complete correction of Fallot tetralogy]. AB - Infundibulo-pulmonary aneurysm is a rare complication of complete correction of Tetralogy of Fallot and its recurrence has not been previously reported. A girl with Tetralogy of Fallot with two small pulmonary branches underwent complete correction at 3 years of age with widening of the infundibulum, the pulmonary annulus and artery with a pediculated pericordial path. Five years later, the left parasternal systolic murmur increased in intensity due to an infundibulo pulmonary aneurysm and severe stenosis of the bifurcation of the pulmonary artery confirmed by echocardiography and catheterisation. The child was reoperated with resection of the aneurysm and widening of the pulmonary tract and its two branches with a Dacron patch. Three years later, the aneurysm and pulmonary stenoses recurred and required percutaneous angioplasty and stenting. The inadequacy of the result led to a further surgical procedure. PMID- 9749221 TI - [Value of the spiral angio-scanner with three-dimensional reconstruction in the surgical strategy of unifocalization. Apropos of a case]. AB - One of the difficulties of surgical treatment of pulmonary atresia with patent septum by unifocalisation resides in the accurate diagnosis of the different collateral vessels to the lung in order to optimise the surgical approach: anterior or posterolateral thoracotomy, and to determine the type of operation: one or two stages repair. Conventional angiography, even using different views, cannot always give an accurate representation of the anatomy of the different collateral vessels, especially their relationship to the bronchial structures. The authors report the contribution of spiral angioscanner with three dimensional reconstruction in the determination of the operative strategy of a case of pulmonary atresia with patent septum. PMID- 9749222 TI - [Vigilance about material: an ethical necessity]. PMID- 9749223 TI - [Hospital results of angioplasty in multiple coronary vessel disease in 1990-1991 and 1994-1995. What conclusions should be drawn about the improvement of results with respect to the relevance of data of randomized trials comparing angioplasty and surgery]. AB - The aim of this study was to determine whether advances in angioplasty techniques have improved results in multiple vessel coronary disease and to compare present results with those reported in randomised trials comparing angioplasty and surgery. The hospital results of two cohorts of multivessel coronary patients treated by angioplasty during two different periods were compared (group 1: 1990 1991. group 2: 1994-1995). The first period corresponded to the inclusion period of randomised trials comparing surgery and angioplasty. The patients in group 2 (n = 449) were older than those in group 1 (n = 424), had more triple vessel disease, more severe angina and more previous angioplasty attempts. Moreover, there were more cases of unfavourable lesions. Nevertheless, the clinical success rate was high in group 2 (92% vs 84%; p < 0.001) and the major complication rate (death, myocardial infarction or emergency bypass surgery) was lower (2.9% vs 6.1%; p = 0.02). The main technical difference between the two periods concerned the use of coronary stents (12% vs 8%; p < 0.001). The fact of being in group 2 was identified by multivariate analysis as an independent predictor for clinical success and a lower major complication rate. The authors conclude that, since the publication of randomised trials comparing angioplasty with coronary surgery, the hospital results of angioplasty have significantly improved. This should be taken into account in considering the clinical applications of the results of these trials. PMID- 9749224 TI - [Comparison of echocardiographic automatic border detection and magnetic resonance imaging. Measurements of the left ventricle in normal subjects]. AB - Echocardiographic automatic border detection (ABD) has been the object of several studies with diverging results. The aim of this study was to verify the validity of ABD measurements by comparison with magnetic resonance imaging (MRI). Twenty healthy subjects underwent measurement of end systolic surface (ESA) and end diastolic surface areas (EDA) and fraction of surface variation (FSV), end systolic volume (ESV), end diastolic volume (EDV and ejection fraction (EF). These results were compared with the same parameters measured by cine MRI and a study of the variability of interpretation was performed on the echocardiographic parameters. An ABD analysis was possible in 80% of the study population. The correlations were satisfactory between the EDA and EDV (EDA; r = 0.84; SD = 1.9 cm2; EDV; r = 0.90; SD = 12 ml) with acceptable confidence intervals (CI) (EDA; [ 4.02; 1.19 cm2]/EDV; [26; +7.9 ml]) and an underestimation of ABD values (EDA; 9%/EDV: -10%). With regards to the end systolic measurements, the correlations were not as good (ESA: r = 0.68; SD = 1.5 cm2/EDV: r = 0.59; +12 ml) with a more important measurement error (ESA: -2.05; +3.45 cm2)/EDA: (-9; +27 ml) and an overestimation of the ABD values (ESA; +10%; ESV; +18%). No correlation was observed between the FSV and EF. The intra and inter-observer errors were compared with those of conventional echocardiography (intra-observer error; 10.7 16.9%/inter-observer error; 10.8-16.6%). The authors conclude that ABD has a non negligeable measurement error which limits its application in clinical practice. New transducers, automatisation of gain adjustment and new technologies should improve ABD measurements. PMID- 9749225 TI - [The cardiowest total artificial heart: experience of 29 cases]. AB - The authors studied the outcome of multi-organ failure in 29 patients with terminal cardiac failure and maintained with a Cardiotest total artificial heart whilst waiting for cardiac transplantation. Pre-implantation organ dysfunction was defined by the following criteria; assisted respiration of over 3 days, total billirubin and creatinine levels of over 2 mg/dL, a platelet count of less than 80,000/mL or a prothrombin ratio of less than 50% and central nervous system disturbances. Fourteen patients died during the period of circulatory assistance and 71% of deaths were due to multi-organ failure. Pre-implantation plasma total bilirubin levels were significantly higher in patients who died of multi-organ failure (p = 0.04). Eighty per cent of patients who died of multi-organ failure had at least 3 criteria of organ dysfunction before implantation of the artificial heart compared with only 37% in the other patients (p = 0.04). Finally, systemic vascular resistances before implantation were significantly lower in patients who died of multi-organ failure. The results of this study suggest that multi-organ failures does not develop during the period of circulatory assistance but represents an aggravation of a preexisting morbid condition. This observation should lead to a limitation of the indications of total circulatory assistance in some cases and, above all, to earlier intervention before multi-organ failure becomes irreversible. PMID- 9749226 TI - [Percutaneous coronary angioscopy in the diagnosis of cardiac graft coronary disease: comparison with the results of angiography]. AB - Coronary disease in cardiac transplant patients is a major factor in the limitation of long term survival. The aim of this study was to compare the results of angioscopy with those of coronary angiography performed systematically every 18 months in our center. Twenty-nine patients (31 angioscopies) were assessed 38 +/- 21 months after transplantation. The appearance observed by angioscopy were: 1) normal, 2) yellow pigmentation of the arterial surface, 3) elevated plaque < 50%, 4) elevated plaque > or = 50% stenosis. Angiography was: 1) normal, 2) iregularities of the lumen or < 50% stenosis, 3) > or = 50% stenosis. The films were viewed by two independent investigators. Angioscopy was performed on the left anterior descending artery (N = 35), the left circumflex (N = 24) and the right coronary artery (N = 9). One to three arterial segments were examined per vessel (total of 117 segments: average 3.8 segments per patient). Angioscopy was uniterpretable in 13/117 (11%) of cases. Of the 81 (78%) segments considered normal at coronary angiography, only 55 seemed normal at angioscopy (68%). Of the 23 segments considered to be abnormal at coronary angiography, all were also considered to be abnormal at angioscopy. The authors conclude that coronary angioscopy seems to be more sensitive than coronary angiography for the detection of coronary disease due to chronic rejection. Prospective studies are required to determine whether the infra-angiographic angioscopic lesions correspond to earlier stages of coronary disease of the cardiac graft. PMID- 9749227 TI - [Calcium inhibitors in the management of hypertensive patients on cyclosporine]. AB - Cyclosporine (CsA) is an immunosuppressor widely used in all postoperative transplantation protocols. Nephrotoxicity and hypertension are the major secondary effects of cyclosporine. The CsA acts on two essential regulatory mechanisms of the blood pressure: the extracellular volume and the systemic vascular resistances. The incidence of hypertension in patients treated with CsA varies from 10% to 80% in the literature. In view of two main mechanismes implicated in CsA-induced hypertension the most logical therapeutic approach would be to use diuretics and calcium inhibitors. The major drawback of diuretic therapy is the risk of hyperuricaemia and attacks of gout, the predisposition to which is already increase by treatment with CsA. In addition, renal failure may rarely be observed. The calcium channel blockers are the drugs of choice in the treatment of CsA-induced hypertension. Not only are they effective in decreasing the blood pressure, but they also have a major advantage of having a nephroprotective effect against CsA. Cyclosporine is metabolised in the liver by cytochrome P450 3A4 dependant enzymes. Many pharmacological interferences have been described with this drug and other pharmacological agents. This type of interaction has been described with certain calcium inhibitors which inhibit hepatic and digestive degradation of cyclosporine and therefore increase its plasma concentrations. The choice of calcium inhibitors should be based on criteria of efficacy and tolerance and on the drug's pharmacokinetics. The interaction between cyclosporine and some calcium inhibitors exposes the patients to the risk of overdosage when treatments is instituted and of underdosage when the treatment is withdrawn. PMID- 9749228 TI - [Aorto-coronary dysplasia and pseudoxanthoma elastica]. AB - The authors report the case of a 50 year old man with pseudowanthoma elastica with a history of myocardial infarction and severe aortic regurgitation. Angiography showed multiple coronary artery aneurysms and aneurysmal dilatation of the aortic annulus. The outcome after triple coronary bypass surgery with aortic valve replacement in a valved Bentall conduit was favourable. Pseudoxanthoma elastica is a rare condition in which the prognosis depends on the degree of vascular involvement. In this context, coronary artery aneurysms and aneurysmal dilatation of the aorta are rare complications. PMID- 9749229 TI - [Left ventricular aneurysm in human immunodeficiency virus infection: a case report]. AB - The authors report the case of a 30 year old man with a left ventricular aneurysm who was seropositive to HIV 1 and HIV 2. The patient was stage IVC 1 (AIDS related complex) by the "Center for Disease Control" classification. The clinical presentation was pyrexia, loss of weight, micropolyadenopathy and cardiac failure. The electrocardiogramme showed low voltage in the peripheral leads with a QS morphology in S2, S3 and aVF and abrasion, of the R wave in the precordial leads. Doppler echocardiography demonstrated a large left ventricular aneurysm with a wide neck. Despite treatment with a diuretic, angiotensin converting enzyme inhibitor and anticoagulants, the patient died suddenly. Autopsy confirmed the wide necked left ventricular aneurysm. This would appear to be the first report of this form of cardiac disease during HIV infection. However, a simple coincidence of the two pathologies cannot be excluded. PMID- 9749231 TI - [Good usage of randomization]. PMID- 9749232 TI - [Diagnostic value of ST depression corrected for heart rate in the post-exercise recovery period]. AB - This study assessed the diagnostic value of two new electrocardiographic criteria of coronary artery disease: the ST/HR index and the slope of the linear relationship between ST segment changes and the heart rate during the first three minutes of the post-exercise recovery period. These two criteria were compared to the standard criteria (> or = 1 mm horizontal or descending ST depression or > or = 2 mm ascending ST depression) to Detrano's ST/HR exercise index (> 1.6 microV/bpm in coronary patient), the exercise ST/HR slope (> or = 2.4 microV/bpm in coronary patients) and the exercise recovery loop (clockwise in normal and anticlockwise in coronary patients) in 88 subjects investigated for suspected coronary artery disease who underwent a computerised exercise stress test and coronary angiography (25 single vessel, 21 double vessel, 20 triple vessel disease; 22 with no significant coronary disease). The ROC identified thresholds of abnormality of the ST/HR recovery index at > or = 2.1 microV/bpm and of the ST/HR recovery slope at > or = 2.52 microV/bpm. Global comparison of the areas under the ROC showed the diagnostic superiority of the exercise ST/HR indices (0.96) over the standard criteria (0.92) and recovery indices (0.86) but without statistically significant values (p = 0.65 and p = 0.15 respectively). The ST/HR index and slope during recovery identify coronary disease with a diagnostic accuracy of 80% and 77% respectively which is similar to that (84%) of the standard ST criteria. The exercise-recovery loop was less accurate (64%). PMID- 9749230 TI - [Renovascular hypertension due to renal artery thrombosis]. AB - The authors report the case of a 74 year old woman admitted to hospital for severe hypertension with unilateral renal artery thrombosis. Recanalisation of the renal artery was obtained by transluminal angioplasty leading to rapid control of the hypertension. Dynamic renal scintigraphy with MAG 3 confirmed the viability of the kidney distal to the thrombosis and, secondarily, the functional recovery of the affected kidney. PMID- 9749233 TI - [Comparative efficacy and risks of low molecular weight heparins and thrombolysis in massive pulmonary embolism without cardiogenic shock]. AB - The aim of this retrospective study was to assess pulmonary reperfusion by scintigraphy, the risks of recurrent embolism and of bleeding complications at the 7th day and 3rd month in 2 groups of patients admitted to hospital for massive pulmonary embolism without cardiogenic shock treated by intravenous thrombolysis (Group I) and by subcutaneous low molecular weight heparin (Group II) paired by Miller's index. The basal characteristics of the two groups, each comprising 31 patients, were comparable with respect to the severity of the pulmonary embolism with an average global scintigraphic defect of 40.6 +/- 13.5% in Group I and 39 +/- 13.7% in Group II. The scintigraphic changes at the 7th day were comparable with a relative improvement of 55 and 51% respectively and at 3 months of 74% in both groups. There was no significant difference in terms of recurrence of embolism (3 versus 0% at the 7th day and 3% in each group at 3 months) or of bleeding complications (13 and 10% at the 7th day and 10 and 6% at 3 months respectively). Low molecular weight heparin seems to be as effective as intravenous thrombolysis for the treatment of massive pulmonary embolism without shock. This result requires confirmation by a large scale prospective randomised trial. PMID- 9749234 TI - [Influence of time (4 years) on the results of programmed ventricular stimulation]. AB - The reproducibility of programmed ventricular stimulation has been previously demonstrated for periods of a few hours to several months. It has not been studied over longer intervals. The aim of this study was to assess the reproducibility of the method at long-term (> 2 years). Forty-six patients with underlying cardiac disease underwent two programmed ventricular stimulations in the absence of antiarrhythmic treatment at intervals of 2 to 6 years (mean 4 years). None of the patients had myocardial infarction or cardiac surgery during this period. The protocol was identical: up to 3 extra-stimuli were delivered in the two right ventricular sites over 3 cycles. Twenty-eight patients had inducible sustained monomorphic ventricular tachycardia during the first investigation (Group I): the investigation was negative in the remaining 18 patients (Group II). During the second investigation, 26 of the 28 patients in Group I had inducible ventricular tachycardia, the rate of which decreased from 206 +/- 50 bpm to 196 +/- 54 bpm. The induced ventricular tachycardia was slower in 15 patients and faster in 5 patients. The mode of induction was different in 12 cases. In Group II, 4 patients (22%) had inducible sustained ventricular tachycardia at the second investigation. The authors conclude that the reproducibility of programmed ventricular stimulation remains good in the long term in subjects within inducible tachycardia, demonstrating the stability of the arrhythmogenic substrate; the frequency of this tachycardia is generally slower. In subjects with an abnormal initial investigation who became symptomatic, it may be useful to repeat programmed ventricular stimulation. PMID- 9749235 TI - [Signal averaging electrocardiography in chronic alcoholism]. AB - Cardiovascular death is the main cause of mortality in chronic alcoholics, perhaps due to a pro-arrhythmogenic effect of alcohol associated with infraclinical myocardial lesions. The authors investigated prospectively 41 patients (average age: 49.7 years) who were chronic alcoholics but had no acute alcoholic episodes for cardiac disease (ECG, signal averaging for late ventricular potentials, echocardiography and Holter ECG monitoring) and hepatic disease (liver biopsy). The history of alcoholism was 14 +/- 9 years, the quantity of alcohol ingested before they stopped drinking being 89 +/- 31 grammes/day. Thirty per cent of patients displayed 2 or 3 criteria of late ventricular potentials (LP). The authors demonstrated a correlation between the daily quantity of alcohol consumed before stopping drinking and the duration of the filtered QRS complex (p = 0.02). Moreover, the frequency of fatty infiltration found on liver biopsy, greater in alcoholics with LP (35% versus 19%, p = 0.025) correlated with the amplitude of the last 40 ms of the average QRS (p = 0.0485), with the duration of potentials of less than 40 microvolts (p = 0.05) and, above all, with the number of criteria of LP (p = 0.02). Finally, the presence of LP was also related to the following biological abnormalities: GGT (p = 0.027), ASAT (p = 0.046), ALAT (p = 0.039). The ECG abnormalities may reflect early infra-clinical myocardial lesions secondary to cellular metabolic abnormalities perhaps analogous to the fatty hepatic changes. However, the prognostic value of these signal-averaging ECG abnormalities remains unknown. PMID- 9749236 TI - [Arrhythmogenic effects of extracorporal lithotripsy in desynchronized mode with a later generation lithotriptor]. AB - Extracorporal lithotripsy has transformed the treatment of renal stones. However, the shock waves can generate arrhythmias which may be severe. To prevent them, the shocks were synchronised to the absolute refractory period of the cardiac cycle with a corollary, an increase in the procedure time. The latest generation of lithotriptors have a number of technical improvements allowing desynchronisation but with a high theoretic risk of arrhythmias. The aim of this study was to assess the arrhythmogenic effects of desynchronised shocks by Holter ECG monitoring during the procedure. This was a prospective randomised study: 25 patients with no previous cardiovascular history were included: 15 men and 10 women with a mean age of 45 years. The originality of this study was the delivery of shock waves in two phases (synchronised versus desynchronised) in a random fashion, the order being unknown to the cardiologist interpreting the recordings. In the synchronised mode, none of the patients developed an arrhythmia. On the other hand, in the desynchronised mode, hyperexcitability was observed in 7 patients, supraventricular in 6 cases (atrial extrasystoles and atrial doublets) and ventricular in 4 cases (VES, ventricular doublets and 1 unsustained ventricular tachycardia). These arrhythmias were asymptomatic and regressed spontaneously. The authors conclude that although they are arrhythmogenic, desynchronised procedures with the latest lithotriptors are acceptable, provided that the patients undergo a cardiological examination beforehand to identify high risk patients and that there is adequate monitoring during the procedure in the presence of an anaesthetist. PMID- 9749237 TI - [Frequency and nature of atheroma at the site of spasm of angiographically normal or subnormal coronary arteries: an endocoronary ultrasonographic study]. AB - Anatomical studies suggest that sites of coronary spasm are subject to early atherosclerosis. Coronary angiography is unable to confirm the lesions or provide information about their nature. On the other hand, endocoronary ultrasound is able to identify and, it is hoped, to determine the frequency and composition of the lesions. Nineteen patients with chest pain and angiographically normal or subnormal coronary arteries were included in a prospective study (16 men and 3 women: average age 53 +/- 10 years). Four patients had spontaneous spasm and in the other 15, spasm was induced by intravenous injection of ergometrine (6 micrograms/kg). After countering the spasm with isosorbide dinitrate, the site of spasm and adjacent segments were examined by endocoronary ultrasound. Localised vasospasm which was stenotic in 14 cases and obstructive in 5 cases, was observed. The ECG was unchanged in 4 cases and showed ST-T segment changes in 15 cases. The artery affected was the left anterior descending in 10 cases, the left circumflex in 2 cases and the right coronary in 7 cases. A plaque of atheroma, defined as significant intimal thickening, was detected in 18 out of the 19 cases. This atheroma was classified as soft in 17 cases and hard in one case. The authors conclude that vasospasm is not only associated with a plaque of atheroma, nearly always suspected at coronary angiography, but also its composition is nearly always soft (lipidic) from ultrasonographic data. PMID- 9749238 TI - [Low-dose dobutamine echocardiographic assessment of reversible contractile dysfunction (myocardial stunning) after myocardial infarction]. AB - Low dose (5 to 10 micrograms/min) dobutamine echocardiography was used to predict the presence of reversible contractile dysfunction (myocardial stunning) after myocardial infarction successfully revascularised in the acute phase of primary angioplasty. The investigation was undertaken in 40 patients, 4 +/- 1 days after inaugural myocardial infarction. The left ventricle was divided into 16 segments. Viable myocardium was diagnosed when the initial regional wall motion score decreased by at least 2. Resting echocardiography was performed at 2 months to evaluate the effective recovery of regional wall motion (myocardial viability). The presence of contractile reserve was documented by dobutamine echocardiography in 18 patients (45%). The sensitivity, specificity and positive and negative predictive values of dobutamine echocardiography for the diagnosis of myocardial viability were 82, 83, 78 and 86% respectively. The negative predictive value was high in all dysnergic segments (86%). The positive predictive value of the investigation was however higher in hypokinetic than in akinetic segments (73 vs 21%; p < 0.05). The recovery of regional wall motion during follow-up was statistically related to higher initial left ventricular ejection fraction (p < 0.04), the presence of angiographically documented collateral circulation before revascularisation (p = 0.007), the contractile response to dobutamine (p = 0.0001) and was observed more frequently in hypokinetic than in akinetic segments (p < 0.05). These results show that low-dose dobutamine echocardiography is a sensitive and specific investigation for predicting irreversible myocardial damage after successful primary angioplasty in acute myocardial infarction. However, even in the absence of residual coronary stenosis, the presence of viable myocardium is only identified specifically in hypokinetic segments. PMID- 9749239 TI - [Heart rate variability and vagal syncope in the young adult]. AB - Heart rate variability is a sign of sympathetic activity. The authors compared two study populations of young males aged 19 to 30 years: population T comprised 15 healthy volunteers who had two negative tilt tests, one under basal conditions and the other after a bolus of isoproterenol; population S comprised 12 patients without cardiac or other disease, who were followed up for malaise and in whom the basal tilt test was positive, confirming the vagal origin of syncope. Temporal and spectral (total power, low frequency 0.04-0.15 Hz, hight frequency 0.16-0.40 Hz) data was obtained concerning heart rate variability from 24 hour Holter monitoring. The main difference between the two study populations was in the temporal data over 24 hours especially with respect to the heart rate (T = 73.5 +/- 6.9; S = 65.4 +/- 6.2/min; p = 0.004) and the percentage of successive R R intervals varying by more than 50 ms (PNN 50) (T = 20.2 +/- 8.3%; S = 30.7 +/- 10.2%; p = 0.024). At night, the lowest SDANN/5 (standard deviation of RR intervals over periods of 5 minutes) were observed in group S (67.2 +/- 16.7 ms vs 87.3 +/- 24.4 ms; p = 0.026). No statistically significant differences between the two groups was observed in the spectral data. The temporal data of heart rate variability on Holter ECG monitoring over 24 hours could therefore have a good predictive value of the vagal origin of syncope in young adults. PMID- 9749240 TI - [At the heart of Internet]. AB - Internet combines features of both a "coffee shop" and a "very large library". It is different from these two institutions in the innovating approach to information and its communication. The information is obtained by navigators, who recover documents dispersed in the four corners of the World Wide Web. The hypertext links between these documents explain the originality of the Web. They are a permanent invitation to look "somewhere else" to enrich the original site. Electronic messengers facilitate the exchange of information and documents. Room for discussion is available for all in the forums and to a well identified community in the lists of users. Internet is a decentralised and anarchic system. In order not to get lost, the sign posts must be followed. To avoid being a nuisance, there are rules of good conduct. You must not expect too much in order not to be disappointed: absolute security (with the risk of not having access) and perfect adequation of content (because of the risk of the loss of freedom of expression). PMID- 9749241 TI - [Double stenosing valve disease after mediastinal radiotherapy: a case report of a 31 year-old man]. AB - Cardiac complications of radiotherapy for cancer, especially lymphoma and breast cancer, are well documented. The three tunics of the heart can be affected. However, valvular disease is rare and, when present, is usually regurgitant. Stenosis is very rare. The authors report the case of a 31 year old man who developed double mitro-aortic valvular stenosis 20 years after mediastinal radiotherapy associated with aortic regurgitation, right coronary stenosis and inflammatory epicardo-pericarditis with effusion. Surgery was undertaken and associated double aortic and mitral valve replacement and right coronary by pass grafting. PMID- 9749242 TI - [Multiple myxoma of the right ventricle with infiltration of the pulmonary infundibulum and arteries]. AB - The authors report a case of multiple myxoma of the right ventricle treated surgically. One of the myxomas infiltrated the right ventricular ourflow tract, the main pulmonary artery and its branches. This case underlines the difficulty of radical resection of infiltrating cardiac myxomas. PMID- 9749243 TI - [Myocardial infarction by complete thrombosis of the left main coronary artery: emergency treatment with angioplasty with implantation of a coronary stent and follow-up at one year: a case report]. AB - Acute occlusion of the left main coronary artery is usually responsible for cardiogenic shock, severe arrhythmias or sudden death. Despite the widespread use of emergency coronary angiography in acute myocardial infarction, occlusion of the left main coronary artery is rarely observed and its treatment remains controversial. The authors report the case of a young man with a previous history of radiotherapy for Hodgkin's disease, admitted for acute myocardial infarction due to complete thrombosis of the left main coronary artery treated as an emergency by percutaneous transluminal angioplasty and implantation of a Palmaz Schatz stent. There were no complications of the procedure and the patient was asymptomatic one year later. PMID- 9749244 TI - [Do tight aortic stenoses justify a surgery on a patient older than 75 years]. PMID- 9749245 TI - [Plasma noradrenaline and the prognosis of chronic cardiac failure: a multicenter study]. AB - Plasma noradrenaline is little used in evaluating the prognosis of cardiac failure because of the theoretical necessity of interrupting treatment for a few days before blood sampling. The present study reevaluated the prognostic value of this parameter with blood sampling performed during treatment and then 48 hours after withdrawal of treatment in 192 patients with chronic stable cardiac failure at an advanced stage (64% of patients in Classes III or IV with an average ejection fraction of 28.5 +/- 13.5%). During follow-up (average 43 months) there were 51 deaths and 17 transplants. None of the patients were lost to follow-up. Univariate analysis of 52 variable observers during the initial phase of evaluation found in decreasing order of predictive value for death plasma noradrenaline levels before and after withdrawal of treatment for 48 hours. Serum sodium, age, systolic mean and diastolic pulmonary artery pressures. In multivariate analysis: noradrenaline with or without withdrawal of treatment, hyponatraemia and systolic pulmonary artery pressure. Actuarial survival curves distinguished the following parameters: noradrenaline levels became predictive at concentrations of over 210 pg/mL and there was a significant difference in survival with respect to 4 levels of serum noradrenaline (with or without treatment) > 300 pg/mL, 300 to 600 pg/mL and > 900 pg/mL. This serum noradrenaline measured without withdrawal of treatment (especially angiotensin converting enzyme inhibitors) is a powerful predictor of mortality, carrying a progressively poorer prognosis as the concentration increases. PMID- 9749246 TI - [Extra-coronary atherosclerosis in documented coronary patients]. AB - The prevalence and severity of extracoronary atherosclerosis in 728 patients (572 men and 156 women; average age 59 years) referred for coronary angiography and who had a history of coronary disease for at least 2 years, were assessed by ultrasonography. This population was divided into 3 groups: Group I, 115 patients without lesions at coronary angiography: Group II, 76 patients with mild coronary stenosis ang Group III, 537 patients with at least one severe coronary artery stenosis, a group which included 294 cases of single vessel disease. The authors observed a strong correlation between the presence, severity and diffusion of the coronary artery disease and ultra sonographic signs of peripheral arterial disease: the frequency increased regularly from 45% in Group I to 88% in patients with triple vessel disease in Group III. About two thirds of patients in Group III had carotid or lower limb atherosclerosis and half of them had atherosclerosis or aneurysm of the abdominal aorta. Severe peripheral lesions were not common but all the aortic aneurysms were observed in Group III. Similarly, simultaneous disease in all three peripheral arterial territories ranged from 12% in Group I to 51% in Group III. The risk of finding peripheral arterial disease was increased in patients with coronary artery disease compared with normal subjects. For each peripheral localisation, the risk was two-fold in cases of mild coronary disease and three or four-fold in patients with triple vessel disease in whom the risk of finding at least one severe peripheral lesion was multiplied by ten. The authors conclude that the prevalence and severity of ultrasonographic peripheral atherosclerosis in documented coronary patients was closely related to the presence, severity and diffusion of the coronary lesions. PMID- 9749247 TI - [Coronary angiography by a radial artery approach: feasibility, learning curve. One operator's experience]. AB - The aim of this study was to assess the feasibility of the radial artery approach for coronary angiography in a standard population of presumed coronary patients and to continue the assessment for a sufficiently long period of time to perfect the technique, evaluate the learning curve and prepare a randomised comparison with the femoral approach. The radial artery was used for coronary angiography in 800 patients after exclusion of about 25% of patients, mainly because of a negative Allen's maneuver. With the exception of acute myocardial infarction, there was no selection based on symptoms and transradial catheterisation was attempted irrespective of age, sex, weight or height. The representative nature of the study population was confirmed by the results of the procedure (normal: 20%, single vessel disease: 30%, double vessel disease: 26%, triple vessel disease: 18% and left main disease: 5.4%). The right radial artery was used in 94% of cases. Successful radial puncture/catheterisation was obtained in 97% of cases: 100% of left coronary arteries and 99% of right coronary arteries were catheterised, the left ventricle in 98% of cases, the internal mammary arteries in 100%, and venous bypass grafts in 95%. The average duration of the whole procedure was 19 +/- 9 minutes. This decreased regularly with operator experience and judicious choice of catheters. The best choice seemed to be a single catheter for both coronary arteries, either an Amplatz or a Champ catheter. There were two probably avoidable coronary complications and two transient neurological events but no clinically significant vascular complication. The radial artery seemed to be a good approach for routine coronary angiography and may now be compared with the femoral approach. It should help expand the practice of ambulatory coronary angiography. PMID- 9749248 TI - [Temporal and spectral analysis of heart rate variability in primary dilate cardiomyopathy: evaluation by case control study]. AB - Temporal and spectral analysis of heart rate variability over 24 hours (HRV) was undertaken in 89 patients with primary dilated cardiomyopathy (DCM) confirmed by left ventriculography with normal coronary angiography and compared with 60 control subjects. The left ventricular ejection fraction was 35 +/- 12% in the DCM patients (71 men and 18 women: age 51 +/- 11 years). Clinical signs of cardiac failure were observed in 66% of patients, requiring medication in 62% of patients (diuretics: 47%, digitalis: 45% and ACE inhibitors: 33%). The HRV was significantly lower in the DCM patients than in the control group, even in the absence of clinical signs of cardiac failure. The global HRV was correlated to left ventricular fractional shortening (r = 0.5, p < 0.001) and peak oxygen consumption on exercise (r = 0.56, p < 0.01), but was independent of the degree of left ventricular dilatation, pulmonary capillary pressure and cardiac index. It was not significantly different in cases of sustained or non-sustained ventricular tachycardia on Holter ECG or in cases with late ventricular potentials on signal averaging of the surface ECG. During follow-up of 51 +/- 35 months, patients with decreased HRV on the global indices and those reflecting sympathetic activity had a much higher risk of cardiovascular death and of cardiac transplantation (p < 0.01). The authors conclude that HRV is decreased in DCM. This is mainly related to the degree of left ventricular dysfunction and is independent of the ventricular arrhythmogenic substrate. The HRV may also identify subgroups of patients with DCM at high risk of cardiovascular death or of haemodynamic decompensation requiring cardiac transplantation. PMID- 9749249 TI - [A prospective comparative study of coronary angiography and endocoronary ultrasonography in the detection of coronary lesions after cardiac transplantation]. AB - Coronary angiography is the reference method for the detection of coronary disease of the cardiac grafts which threatens the long-term prognosis of cardiac transplantation. The primary results of treatment for slowing, stabilising or even improving coronary transplant disease are encouraging and make necessary the development and evaluation of reliable diagnostic methods. The authors undertook a prospective study of 48 asymptomatic patients with normal graft wall motion between January 1995 and March 1997 to compare the results of coronary angiography and endocoronary ultrasonography. The patients had been transplanted in the 10 years preceding the study. The results of the two methods were concordant in 33 cases (69%) (NS), for the confirmation (9 cases) or the information of coronary transplant disease (24 cases). The results were contradictory in 15 cases (31%): in 12 cases, endocoronary ultrasonography showed signs of coronary disease whereas the coronary angiography was estimated to be normal: in the remaining 3 cases, coronary angiography was abnormal but no signs of coronary disease were found on endocoronary ultrasonography. The specificity of coronary angiographic detection was 89% and therefore very satisfactory, but its sensitivity (43%) was poor. In addition, endocoronary ultrasonography allows analysis of the extension of coronary lesions to unstenosed segments, the quantification of intimal thickening. Therefore, endocoronary ultrasonography should become the reference investigation for coronary disease of cardiac transplants. PMID- 9749251 TI - [Outcome of type I acute aortic dissection operated after 70 years of age. A retrospective study of operated dissection of the aorta in the over 70 years old]. AB - The aim of this study was to assess the perioperative mortality and long-term outcome of Type I dissection of the aorta operated in patients over 70 years age. Of the 87 dissections of the aorta operated between 1988 and 1995, 19 concerned patients aged 71 to 79 (average 74.1 +/- 2.4 years). The procedure was replacement of the ascending aorta with gluing of the false lumen in call ases. Two patients also underwent aortic valve replacement and three also had replacement or repair of the aortic arch. Eleven patients (56%) had circulatory arrests lasting an average of 33 minutes (10-86 minutes). The mortality rate at 30 days was 31.5% (6/19): 3 deaths were due to bleeding, 1 to mesenteric infarction, 1 to cardiac arrhythmia and 1 to a cerebrovascular accident. The morbidity was 53%, mainly due to neurological complications, chest infection and renal failure. After an average period of 36.8 months (range: 3 to 75 months) with no patient lost to follow-up, the late mortality was 23% (3/13), giving actuarial survival rates at 1.5 and 6 years of 63%, 47.5% and 32%, respectively (including the operative mortality). Or the survivors, 9 were in NYHA Classes I II and 1 in class III. One patient developed a hemiparesis. The authors conclude that, despite high mortality and morbidity at 30 days, long-term survival and its quality are arguments in favour of surgical management, even in elderly patients. PMID- 9749250 TI - [Acoustic quantification of right ventricular dimensions and systolic function]. AB - In view of the important prognostic significance of right ventricular systolic function, there have been many non-invasive studies of this subject. The majority of these studies have been limited by difficulties in modelisation of this geometrically complex cardiac chamber. Automatic border detection by acoustic quantification based on the back scatter of ultrasound provides a "direct" method of analysing right ventricular dimensions and functions. The authors undertook a prospective study of 34 patients to evaluate the reliability of this technique in measuring the surfaces and fractional shortening of the right ventricle. The feasibility was 92%. The correlation coefficients between the manual two dimensional technique and automatic border detection were 0.81 for the end diastolic surface areas, 0.85 for the end systolic surface areas and 0.74 for the fractional shortening. Compared with the isotopic ejection fraction, the correlation coefficient was 0.73. The authors conclude that acoustic quantification is a feasible and reliable technique of measuring right ventricular dimensions and its contractile function. PMID- 9749252 TI - [Reproducibility of heart rate variability in the chronic phase of myocardial infarction]. AB - Evaluation of heart rate variability is a common method of assessing autonomic nervous system function and its effects on heart rate in different conditions. The reproducibility of the technique is not known in the chronic phase of myocardial infarction. The aim of this study was therefore to assess the reproducibility of the measurement in 54 subjects who were clinically stable with no change in treatment at a distance from acute or semi-recent (> 2 years) myocardial infarction, after an interval of one month. The temporal and spectral analysis of heart rate variability included measurement of the standard deviation of the normal RR intervals (SDNN), on the mean heart rate, the percentage of RR intervals greater than 50 ms than the adjacent interval (pNN50), the coefficient of variability (CV), the square root of the differences between successive RR intervals (rMSSD), the power of low frequencies (LF) and high frequencies (HF) and of the fractional spectral power (LF/HF). No significant changes in these parameters were observed. Analysis of individual variations showed that the heart rate was the most stable parameter: for evaluation of vagal tone, the rMSSD showed less variability than the pNN50 and HF. The presence of cardiac disease did not influence these results. The authors conclude that parameters of evaluation of heart rate variability in temporal and spectral analysis are globally reproducible in stable subjects. However, individual values may change from one measurement to another. Nevertheless, abnormal variability is constantly observed at the second investigation and, similarly, normal variability also remains unchanged. These individual variations suggest that, for the demonstration of change in these parameters of variability with treatment, large population groups must be studied. PMID- 9749253 TI - [Reversible graft dysfunction after cardiac transplantation]. AB - The authors report 3 cases of major graft dysfunction after cardiac transplantation which recovered completely with biventricular mechanical assistance in 4 to 8 days. All three cases were primary biventricular graft failures in patients with normal preoperative pulmonary resistances. These early dysfunctions (with no signs of myocardial infarction on electro- or echocardiography and in the absence of abnormal increased peri-operative enzyme levels) associated with total functional recovery conforming to the definition of the phenomenon of myocardial stunning. These results argue in favour of aggressive management of primary graft dysfunction. PMID- 9749254 TI - [Spontaneous rupture of the ascending aorta: a diagnostic and therapeutic emergency. Report of 2 cases]. AB - The authors report two cases of spontaneous rupture of the ascending aorta complicating atheromatous disease but without traumatic or infectious factors. The clinical presentation is very similar to that of dissection of the aorta. The diagnosis is based on non-invasive and invasive investigations showing localised abnormalities of the aortic wall suggestive of localised dissection and haemopericardium. Surgery shows transverse tearing of the aortic wall without dissection of the media. It is a surgical emergency, the deceptive presentation of which should not be missed. PMID- 9749255 TI - [Left atrial myxoma and coronary embolism]. AB - The authors report the case of a 49 year old man referred for a preoperative evaluation of a left atrial myxoma. During the hospital admission, the patient had an episode of sudden anginal chest pain associated with electrocardiographic changes of anteroseptoapical myocardial infarction. Emergency coronary angiography showed occlusion of the middle segment of the left anterior descending artery whereas it had been absolutely normal on the coronary angiogramme performed a few days beforehand. The diagnosis of coronary embolism from the left atrial myxoma was made and an emergency coronary angioplasty was performed followed by surgical ablation of the tumour. PMID- 9749256 TI - [Asymptomatic acute cytolytic hepatitis due to fluindione and associated with resistance to treatment: a case report and review of the literature]. AB - The authors report a case of asymptomatic acute cytolytic hepatitis due to fluindione (Previscan) associated with relative and selective resistance to this drug prescribed as a relay of heparin therapy for deep venous thrombosis following immobilisation of a sprained ankle in a 22 year-old patient. The main complications of oral anticoagulants are bleeding. Very severe immuno-allergic complications, especially hepatic, have been described with phenindione therapy. A review of the literature revealed a very low incidence of fluindione-induced hepatitis (7 cases), only two of which were fully documented since the commercialization of this molecule in 1971. No fatalities were reported: resistance to the treatment seemed to be associated. The exact mechanism (toxicity or immuno-allergy) has not been determined although the fact that fluindione is one of the indanedione family is in favour of the latter hypothesis. PMID- 9749257 TI - [Thrombosis of the inferior vena caval and right atrium in amoebic abscess of the liver]. AB - Amoebic abscess of the liver is sometimes complicated by deep venous thrombosis but extension to the right atrium is rarely observed. The authors report the case of inferior vena caval thrombosis extending to the right atrium in a case of amoebic hepatic abscess. The patient was treated initially by antibiotherapy with metronidazole associated with intravenous anticoagulation. Rapid extension of the thrombus despite this treatment led to the initiation of thrombolysis. There were no embolic complications and the outcome was good. Apart from the rarity of this complication, this case poses the problem of the management of these patients. No previous reports of the use of thrombolysis were found in the medical literature. In the light of previous publications and the present case, the authors suggest investigation by CT scanning, echocardiography and venous Doppler ultrasonography in all cases of hepatic amoebic abscess. PMID- 9749258 TI - [Let us face these challenges together!]. PMID- 9749259 TI - [Evaluation of echo-guided pericardiocentesis in cardiac tamponade]. AB - Between April 1982 and December 1995, 78 consecutive patients with an average age of 57 +/- 13 years underwent echo-guided pericardiocentesis in the intensive care unit for poorly tolerated pericardial effusions. The patients were admitted to the cardiology departments of Ambroise-Pare Hospital at Boulogne (n = 44). Gilles de-Corbeil Hospital at Corbeil-Essonnes (n = 31) and Val-de-Grace Hospital in Paris (n = 3). The underlying aetiologies were malignant disease (n = 31), idiopathic (n = 13), post-surgery (n = 7), infection (n = 7), autoimmune (n = 6), post-radiotherapy (n = 6), post-myocardial infarction (n = 3), chronic renal failure (n = 3) and coagulation defects (n = 2). Pericardial puncture was undertaken by the subxiphoid (n = 77) or left parasternal (n = 1) approaches under guidance of echocardiography. Intra-pericardial contrast was used to verify the position of the catheter. The average volume of liquid drained was 580 +/- 390 mL. After pericardiocentesis, continuous drainage was continued in 17 patients for an average duration of 63 +/- 29 hours. The total average volume was 750 +/- 330 mL. The major complications were a) three deaths during the puncture, not caused by the procedure after post-mortem study, b) ten right ventricular punctures with no consequences in 9 cases, c) two cases of shock, one of which was due to a pre-existing septicaemia of pulmonary origin, d) two non-sustained ventricular arrhythmias. The minor incidents were six vasovagal syndromes during the procedure and four paroxysmal supraventricular arrhythmias. Emergency surgical drainage was required (n = 3) for a failed procedure and late surgical drainage (n = 12) for persistence or recurrence of the effusion. No surgical drainage was required in the 17 patients placed under continuous aspiration. Echo guided pericardiocentesis is a simple procedure and provides rapid haemodynamic relief in subjects generally in serious condition. Continuous aspiration may help avoid the need for surgical drainage for persistence or recurrence of the effusion. PMID- 9749260 TI - [Long-term results of aorto-bifemoral prostheses in the surgery for atheromatous stenosis of the aortic bifurcation]. AB - Seven hundred patients operated consecutively by the same surgical team for atheromatous stenosis of the aortic bifurcation were followed up for 20 years with only 5 patients lost to follow-up. There were 94.5% of men with a mean age of 58 years. The operative mortality was 2.7% with few deaths due to true cardiovascular causes (0.7% of patients). The secondary mortality was very high with two main causes: cancer (39% of patients) and cardiovascular diseases (37%). Other causes were responsible for only 24% of deaths. The principal complications of the prostheses were: infection (0.6% of operated patients), thrombosis (6.7%), pseudo-aneurysm of the aorta (1.57%) and pseudo-aneurysm of Scarpa's triangle (4%). The benefits of surgery are unquestionable both on terms of survival, as amputation, bed confinement and invalidity are avoided, and in terms of function, as amputation was avoided in 84% of the 32% of patients in Stages III or IV before surgery. Only 5% of patients were amputed during the observation period. Moreover, 79% of survivors had a good functional result at 15 years. Improvement of results depends on better hygienic measures, systematic screening for high risk cancers and a better management of the arterial disease with early treatment of other arterial diseases (coronary, carotid) in order to reduce postoperative and mostly medium- and long-term cardiovascular mortality. PMID- 9749261 TI - [Doppler tissue imaging of pre-ejection left ventricular wall dynamics in normal subjects]. AB - Pre-ejectional left ventricular wall motion has been demonstrated clinically by angiography. Intramyocardial wall velocities generated by cardiac contraction may be measured by Doppler tissue imaging. The aim of this study was to detect pre ejectional wall motion and to analyse its sequencer. A long axis M Mode with simultaneous septal and posterior wall imaging was performed in 11 normal subjects (age 37 +/- 15 years) with velocity analysis between the electrocardiographic Q wave and the onset of ejection by digitised analysis between the electrocardiographic Q wave and the onset of ejection by digitised images with automatic velocity extraction (3.8 ms) along a horizontal subendocardial line. The total duration of the pre-ejectional periods in conventional and Doppler tissue imaging are compared. Oscillatory velocimetric appearances with alternate colours of adjacent bands in each wall and a mirror image between walls was observed. The mean and peak velocities of the first four bands were significantly different between the walls (p < 0.001) as were the absolute values between bands 2 (p < 0.02) and 3 (p < 0.006). The duration of band 2, related to motion mainly towards the center of the ventricular chamber exceeded that of the adjacent bands (septum p < 0.02, posterior wall p < 0.001). The correlation coefficient for total duration of the pre-ejectional period between Doppler tissue imaging and conventional Doppler was 0.83, p < 0.05 for the interventricular septum and 0.76, p < 0.04 for the posterior was. The authors conclude that regional pre-ejectional wall motion can be recorded. During isovolumic contraction, there is motion predominantly towards the center of the left ventricular chamber of the two walls, confirming previous angiographic findings. Its timing suggests that wall motion proceeds the increase in ventricular pressure. PMID- 9749262 TI - [Long-term outcome of a false lumen after surgical correction of type A acute aortic dissection]. AB - The long-term outcome (64.3 +/- 45 months) of 44 patients operated for acute dissection of at least the ascending aorta was assessed by regular clinical examination and annual CT scan. The diameter of the aorta at different levels was measured at each CT scan for all patients. Initially, 7 patients (16%) had acute dissection limited to the ascending aorta; none had a false lumen after surgery. No signs of aneurysmal dilatation were observed during follow-up of these patients. In the 37 other cases (84%) dissection of the aorta extended beyond the innominate artery; the false lumen remained patent distal to the prosthetic tube replacing the ascending aorta in 34 patients (92%). The false lumen was partially thrombosed in 8% of patients, leading to distal emboli in 1 patient. Moderate increases (less than 15 mm) in diameter of the false lumen were observed in 32% of patients; more severe dilatation (over 20 mm) was observed in 12% of patients. The management of dilatation of the false lumen is not standard; it depends mainly on the rate of progression and the clinical consequences. It is hoped that extension of the initial repair to the aortic arch, when the intimal tear is situated in this zone, will reduce the short and long-term progression of the false lumen. PMID- 9749263 TI - [Alcoholic cardiomyopathy and heart transplantation]. AB - The recognition of alcoholic cardiomyopathy in patients with dilated cardiomyopathy is essential as they may regress, at least partially in a relatively short period, with abstention. The clinical history is the key to diagnosis because no other specific feature can identify the cause. Between January 1984 and July 1995, 26 candidates for cardiac transplantation with dilated cardiomyopathy and chronic alcoholism improved after withdrawal of alcohol. None of these patients was placed on the surgical waiting list. Patients with ischaemic cardiomyopathy, valvular disease or previous surgery for valvular hypertensive or congenital heart disease, documented viral myocarditis or connective tissue diseases, were excluded. The diagnostic criterion of chronic alcoholism was a total alcohol consumption of 292 kg and a duration of alcohol abuse of over 10 years. In addition to the clinical features, biological, electrocardiographic, echocardiographic and haemodynamic parameters were analysed. The mean age of the patients was 48 +/- 8 years. There were 25 men and 1 woman. The total mean alcohol consumption was 1,492 kg. The average follow-up period was 63 +/- 41 months. The interval between the onset of symptoms and abstention was 25 months. Haemodynamic improvement was observed in 25 cases. The average interval between alcoholic abstention and recovery was 11.7 months. One patient died suddenly. Improvement of symptoms, decrease of the cardiothoracic ratio and improvement of echocardiographic parameters were statistically significant. The increase in angiographic or isotopic ejection fraction and cardiac index and the decrease in mean pulmonary artery pressures were also statistically significant. These results confirmed the diagnosis of alcoholic cardiomyopathy. Therefore, patients with chronic alcohol abuse and dilated cardiomyopathy must be identified and treated for this problem and not placed on the waiting list for cardiac transplantation unless no improvement is observed after about 3 months of abstention. PMID- 9749264 TI - [Rapid decrease of serum endothelin-1 levels after treatment with ACE inhibitors in chronic cardiac failure]. AB - Circulating endothelin-1, a very strong peptide vasoconstrictor first discovered in 1988, is raised in cardiac failure. This increase contributes to the deleterious effects of cardiac failure. Although specific anti-endothelin drugs are under development in this condition, the effects of more commonly used drugs on circulating endothelin-1 levels are not well known. The aim of this study was to evaluate the effects of ACE inhibitor therapy on plasma endothelin-1 levels in cardiac failure. The plasma endothelin-1 levels were measured in 24 patients (stages III and IV of the NYHA classification), before and after treatment with angiotensin converting enzyme inhibitor (Captopril: test doses, then 75 mg/day). A control group of 10 paired patients was used to shake off the effects of bed rest and hospital salt-free diet. The initial endothelin-1 levels were high but equivalent in the control and study groups: 9.13 +/- 1.87 fmol/mL vs 8.98 +/- 1.92 fmol/mL. Plasma endothelin-1 decreased significantly in the study group 72 hours after beginning ACE inhibitor therapy (7.44 +/- 1.95, p < 0.02) but remained higher than normal (p < 0.01), whereas the values in the control group remained the same. This study demonstrates a decreases in plasma endothelin-1 levels 72 hours after onset of ACE inhibitor therapy in patients with stable severe cardiac failure. This results, already suggested by many experimental studies and certain ancillary clinical trials, could explain some of the beneficial effects of ACE inhibitors in this condition. PMID- 9749265 TI - [QT interval and drugs. Recommendation for drug prescription for patients with long QT syndrome. Clinical Research Group of INSERM 4940 12: Diagnostic Clinic of Congenital Long QT Syndrome]. AB - The genetics of the long QT syndrome are now better understood. However, there is much heterogeneity as three different genes have already been identified affecting the function of sodium and potassium channels. The aim of these recommendations is to draw up a list of drugs which are contraindicated or not recommended in patients with congenital long QT syndromes. The conraindicated drugs are those with which torsades de pointe have already been described. Drugs not recommended are substances which are not electrohysiologically neutral and for which, in view of their modes of action, their metabolism or belonging to a particular therapeutic class, make them very difficult to use in those patients. It is therefore better not to prescribe them whenever possible in this condition. These substances belong mainly to cardiovascular (especially antiarrhythmic), psychotropic, anti-infectious and antiallergic groups of drugs. PMID- 9749266 TI - [Epidemiology of paroxysmal auricular fibrillation]. AB - The epidemiology of paroxysmal atrial fibrillation (PAF) is poorly known because of the difficulties in setting up trials to study this condition. Its biannual incidence is about 2 per thousand. Its prevalence is 1 to 2% in a population of over 65 years of age. Paroxysmal atrial fibrillation is often asymptomatic. An initial episode of atrial fibrillation may remain a single event in a number of cases. Isolated paroxysmal atrial fibrillation progresses to permanent atrial fibrillation in about 20% of cases, usually if there is underlying cardiac disease. Rheumatic valve disease, cardiac failure, hypertension, previous myocardial infarction and cerebrovascular accidents are often associated with paroxysmal atrial fibrillation. Embolic complications are rare if paroxysmal atrial fibrillation is isolated. PMID- 9749267 TI - [Transformation of left bundle branch block into simulated bundle branch block after ablation of the right bundle in a patient with branch-to-branch reentry ventricular tachycardia]. AB - Masquerading bundle branch block associates left bundle branch block in the standard lead and right bundle branch block in the precordial leads. Mr R., 67 year old, was referred for investigation of syncope. He had a history of idiopathic dilated cardiomyopathy (normal coronary arteries; EF: 14%, CI: 2.2 l/min/m2 at later investigations). The ECG showed LBBB with left axis deviation, a PR interval at the upper limits of normal and ventricular premature beats. During observation, he had another syncopal episode and the ECG showed wide complex tachycardia (160 bpm) reduced by external cardioversion. Electrophysiological investigations showed inducible VT due to bundle branch reentry. The HV interval in sinus rhythm was 80 ms. Radiofrequency ablation of the right bundle led to first degree AVB with masquerading bundle branch block with an increased HV interval of 120 ms. The usual facility of ablation of the right bundle branch block is an argument in favour of the hypothesis whereby masquerading bundle branch block is a variety of RBBB with severe conduction defects of the two branches. PMID- 9749268 TI - [Carcinoid heart disease]. AB - Carcinoid cardiac disease is a common complication of metastatic carcinoid tumours. It is characterized by tricuspid regurgitation and pulmonary stenosis. A 68 years old woman with a metastatic carcinoid tumour was admitted to hospital for congestive cardiac failure secondary to severe tricuspid regurgitation. Typical carcinoid lesions of the tricuspid and pulmonary valves were observed at echocardiography. A double valve replacement was performed with a favourable outcome. Postoperative echocardiography showed a significant improvement in right ventricular function. Surgical management of carcinoid valvular heart disease of NYHA Stage III patients is associated with an improved 2 years survival (from 8 to 40%) despite a high operative mortality (about 27%). Cardiac surgery remains the only hope of long-term survival with a spectacular improvement in symptoms. PMID- 9749270 TI - [Therapeutic strategies in acute myocardial infarction]. PMID- 9749269 TI - [Ehlers-Danlos disease revealed during pregnancy through the diagnosis of aortic dissection]. AB - Dissection of the aorta is a serious condition but rare in young women, and occurring during the 3rd trimester of pregnancy. The main risk factors are hypertension and diseases of the connective tissue or of collagen (Marfan's syndrome and Ehlers-Danlos disease). The authors report a case of dissection of the aorta managed in a pluridisciplinary manner by the anaesthetists, cardiologists, obstetricians and cardiothoracic surgeons, which resulted in a favourable outcome for both mother and baby. The diagnosis of Ehlers-Danlos disease was made from the onset and, over a period of 10 years with CT scan and annual echocardiographic follow-up, total replacement of the supra-coronary aorta was performed in several stages. PMID- 9749271 TI - [Physiopathology of myocardial infarction: the acute coronary occlusion]. AB - Rupture of the atheromatous plaque, thrombosis and local vasoconstriction are involved in the genesis of acute myocardial infarction. The vulnerability of the plaque depends on its histological structure. Its fragility is related to the size of the lipid core, the thinness of the fibrous capsule and the inflammatory reaction. External aggression favourites rupture. This triggers both thrombogenesis by bringing the blood cells into contact with thrombogenic subendothelial factors and local vasoconstriction due to endothelial dysfunction. Although rupture of the plaque is an unpredictable event, there is a circadian variability the highest incidence of infarction being between 6 a.m. and midday. Comprehension of the physiopathology of myocardial infarction has opened up new therapeutic approaches which should reduce the incidence of plaque rupture. Prevention is based on stabilisation of the plaque by dietary hygiene, lipid lowering drugs and, maybe, in the future, by local application of antisense oligonucleotides. Finally, anti-aggregant therapy (aspirin or anti-GIIb-IIIa) could prevent the formation or extension of the thrombus. PMID- 9749272 TI - [Antithrombotic agents in acute myocardial infarction]. AB - Coronary thrombosis, which is responsible for myocardial infarction, is a complex phenomenon involving the interaction of the arterial wall, the coagulation system and the platelets. Better understanding of the molecular biology of thrombosis has led to the rapid development of antithrombotic therapy. The limitations of aspirin and heparin have promoted the development of new molecules whose site of action on platelets or at different stages of coagulation are known. Some of them are the object of large scale international trials. Some results have been disappointing such as those with the direct antithrombins: others are promising and in the phase of evaluation, such as the inhibitors of glycoproteins GP IIb IIIa. PMID- 9749273 TI - [Transluminal coronary angioplasty in the acute phase of myocardial infarction]. AB - The objective of the treatment of myocardial infarction is to reestablish patency of the occluded artery as soon as possible. Two methods have been validated: intravenous thrombolysis which is easy to perform, and transluminal coronary angioplasty requiring expensive infrastructures and a skilled medical team but which has a higher success rate of restoring arterial patency. Angioplasty is indicated in cardiogenic shock and cases in which there is diagnostic uncertainty or a contraindication to thrombolysis. In addition, its superiority over thrombolysis has been clearly demonstrated in the following indications: 1) primary angioplasty if proper facilities with an experienced team are available in less than 45 minutes and 2) after failed thrombolysis (rescue angioplasty). The use of stents improves the results of primary angioplasty. Angioplasty and thrombolysis are not rival techniques: the choice depends on local conditions (proximity to a catheterization laboratory with a trained medical team) and the clinical context (presence of "high-risk" criteria). Their association (prehospital thrombolysis followed by immediate angioplasty) is the object of prospective clinical trials. PMID- 9749274 TI - [Comparison of primary angioplasty and prehospital thrombolysis in the acute phase of myocardial infarction. CAPTIM Study Group]. AB - The "Comparison of Primary Angioplasty and Prehospital Thrombolysis in the Acute Phase of Myocardial Infarction" (CAPTIM) trial compares the two best strategies of revascularisation in the acute phase of myocardial infarction. CAPTIM is a prospective, randomised, multicenter, open trial. The patients treated by the emergency ambulance service and presenting the criteria of inclusion were randomised and treated either by rtPA instituted at the pick-up point or taken to hospital for primary angioplasty. The main criterion was composite associating death, recurrent myocardial infarction and cerebrovascular accidents identified at 1 month. These events were validated by a committee independent of the study. The number of required patients was assessed to be 1,200 patients to be recruited in 2 years. CAPTIM started in June 1997 and will finish in June 1999. This study, associating the emergency ambulance service and the coronary care unit of 11 different French cities, should help define the priorities in the actions to be undertaken to optimise the treatment of acute myocardial infarction. PMID- 9749275 TI - [Therapeutic strategies in acute myocardial infarction: antiarrhythmic therapy]. AB - Antiarrhythmic therapy is a special case in the therapeutic strategy of acute myocardial infarction. There are very few controlled therapeutic trials and its use is mainly based on clinical experience rather than on scientific evidence. The most common arrhythmias requiring treatment in acute myocardial infarction are atrial fibrillation, ventricular tachycardia and ventricular fibrillation. There is no evidence to support the use Class I antiarrhythmics. Lidocain may be used in some cases. Similarly, contradictory results have been reported with the use of magnesium salts and the general tendency is not to use this ion in acute myocardial infarction. The most commonly used antiarrhythmic agents are the betablockers and amiodarone. The general principles of treatment should be respected: all antiarrhythmic drugs have negative inotropic effects, apart from digitalis. All antiarrhythmics may have a proarrhythmic effect including digitalis, especially in this clinical context. Whenever possible, continuous intravenous infusions are to be preferred to bolus injections. In addition, and when possible, electrotherapy is preferable to antiarrhythmic drug therapy. Finally, a number of cardiac arrhythmias observed in acute myocardial infarction should be "respected" or treated by electrotherapy but never by antiarrhythmic drugs. PMID- 9749276 TI - [Betablocker therapy in acute myocardial infarction]. AB - A review of the results of randomised trials of the use of betablockers in acute myocardial infarction shows by techniques of meta-analysis that their prescription in the early hours of the acute event leads to a reduction in short term mortality: the reduction of risk is 12% as compared with groups which are not given betablockers. In addition to its impact on mortality, early betablocker therapy is associated with a reduction in the risk of recurrence of myocardial infarction and a proven antalgic effect. However, in patients receiving thrombolysis, the beneficial effect on mortality and recurrence of infarction of intravenous betablockers seems less evident. In daily clinical practice, the prescription of betablockers in the first days of infarction has significantly increased over the last ten years. In the USIK trial carried out in France in November 1995, nearly two thirds of patients received betablocker therapy. In the same study, the prescription of betablockers is associated with a reduction in mortality independent of the classical risk factors and the prescription of angiotensin converting enzyme inhibitors. These results confirm the value of this therapeutic class in the acute phase of myocardial infarction. PMID- 9749278 TI - [XVth non pharmacological treatment of cardiac arrhythmias meeting]. PMID- 9749277 TI - [Treatment with angiotension converting enzyme inhibitors]. AB - Eight randomised clinical trials of angiotensin converting enzyme (ACE) inhibitors versus placebo in myocardial infarction have been published in three years, including over 100,000 patients. High risk myocardial infarction carries a poor prognosis with a 25% death rate at 42 months in SAVE, 23% at 15 months in AIRE and 42% at 37 months in TRACE. This form of myocardial infarction benefits the most from treatment with ACE inhibitors: the relative reductions in risk for the previously mentioned trials were 19%, 27% and 22% respectively, and the absolute reductions in risk of death were 4% at 42 months, 6% at 15 months and 8% at 37 months, respectively. Studies including all forms of myocardial infarction involve populations at lower risk. The effects of ACE inhibitors on mortality are then less obvious: from 7.7% to 7.2% at 5 weeks in ISIS-4, from 7.1% to 6.3% at 6 weeks in GISSI-3, from 9.6% to 9.0% at 4 weeks in CCS-1, corresponding to relative reductions in the risk of death of 7%, 12% and 3% respectively and absolute reductions of the risk of death of 0.5% at 5 weeks, 0.8% at 6 weeks and 0.6% at 4 weeks, respectively. Who should be treated? All patients for 4 to 6 weeks or only high risk patients. When should treatment be started? Some investigators institute treatment from the first day but others think it prudent to wait until the second day. For how long? Four to 6 weeks would be sufficient to counteract ventricular remodelling while the benefits are sustained in the long term. But treatment should be continued long term in high risk patients. Which molecule, which dose and how many times, should it be taken per day? The logical answer is molecules with proven efficacy at the dose given in the clinical trials respecting the number of times indicated per day. PMID- 9749279 TI - [Diagnostic criteria of ventricular tachycardia]. AB - When ventricular tachycardia is very rapid or complicates cardiac disease it must be diagnosed as rapidly as possible so as not to delay treatment. A careful analysis of the surface electrocardiogramme is usually sufficient to distinguish ventricular tachycardia from other-wide QRS complex tachycardias when the widening is due to ventricular aberration. The diagnosis is easier when the start of the tachycardia is recorded or when the sinus rhythm is interspersed with ventricular extrasystoles of the same morphology as that of the tachycardia. Similarly, atrioventricular dissociation is diagnostic of ventricular tachycardia but its negative predictive value is weak. Extreme axial deviation of the QRS complexes, concording morphology in leads V1 or V2 and V6 and the analysis of the QRS complexes in the precordial leads nearly always enables identification of supraventricular tachycardia with aberration. On the other hand, the distinction between other causes of wide QRS complexes (supraventricular tachycardia with preexcitation or intraventricular conduction defects) remains difficult in the absence of a reference electrocardiogramme and the clinical context. PMID- 9749280 TI - [Drug therapy of ventricular tachycardia]. AB - Despite the introduction of new therapeutic techniques such as radiofrequency ablation and the implantable defibrillator, the classical opposition of monomorphic ventricular tachycardia in apparently normal hearts and that arising from documented cardiac disease remains useful. In the first case, treatment is only symptomatic whereas, in the second, lethal progression to sudden death must be prevented. Generally speaking, in chronic post-infarct situations, betablockers are underused although they have been shown beyond doubt to reduce cardiovascular mortality. This is probably explained by the fear of possible haemodynamic decompensation in patients who often have left ventricular dysfunction. Nevertheless, different randomised studies of the use of betablockers in cardiac failure have reported reduced mortality with no serious side effects. The use of beta-blockers is therefore advisable, and possible inpatients with or without sustained ventricular tachycardia and underlying cardiac disease. In cases at high risk of sudden death, amiodarone may be associated. Recent randomised studies (MADIT, AVID), comparing the use of implantable defibrillators with those of antiarrhythmic therapy, have shown better results with the implantable defibrillator. However, in these studies, only about 10% of patients received betablockers in the antiarrhythmic treatment groups. This factor has introduced some doubt as to the real benefit of implantable defibrillators. Therefore, a randomised study comparing the efficacy of betablockers with amiodarone against implantable defibrillators is desirable in order to determine the respective indications of each of these two therapeutic modalities. PMID- 9749281 TI - Mapping techniques and catheter ablation of ventricular tachycardia due to coronary artery disease. AB - In contrast to the high success rates for catheter ablation of focal ventricular tachycardia, radiofrequency ablation for ventricular tachycardia due to coronary artery disease has met with limited to variable success. In performing catheter ablation for ventricular tachycardia due to coronary disease, mapping techniques used to locate the isthmus of the reentrant circuit are crucial. Appropriate use of mapping techniques determines the success rate of the procedure. We classify different methods of mapping into indirect and direct. The indirect methods include analysis of the 12-lead electrocardiogram, endocardial pace-mapping, and analysis of electrograms recorded from different parts of the heart during sinus rhythm. The direct methods include activation sequence mapping and entrainment mapping. The direct methods are more important in evaluating ventricular tachycardia due to coronary disease. While using the technique of entrainment mapping, three criteria are important: a) entrainment with concealed fusion; b) post pacing interval within or less than 10 ms of tachycardia cycle length; c) stimulus-to-QRS interval should equal electrogram-to-QRS interval. A high success rate in terminating the tachycardia is seen if these criteria are met. PMID- 9749282 TI - [Implantable automatic defibrillators in the treatment of ventricular tachycardia]. AB - The implantable automatic defibrillator (IAD) is the treatment of choice of malignant ventricular arrhythmias or those resistant to pharmacological or ablative techniques. However, the small number of implantations in France, the presence of many different models, the cost and sophistication of the latest models explain why IAD remain a poorly known therapeutic method because it is highly specialised and reserved for cardiological departments with a rhythmological interest. Several factors suggest that the number of implantations will increase in the future. Firstly, the techniques of implantation have been considerably simplified and associated with technological improvements of the devices, and, secondly, publication of several trials (MADIT, AVID), even if the conclusions in favour of IAD must be carefully interpreted, will change our methods of management of patients with severe ventricular arrhythmias. The current indications of IAD in ventricular tachycardia and future prospects are discussed in the light of new data. PMID- 9749283 TI - [Vasovagal syndromes]. AB - Most cases of dizziness or syncope referred to the emergency department or to services of internal medicine are caused by vasovagal syndromes. They comprise relative bradycardia with vasoplegia, the cardiovascular response to a neurological stimulus. It is possible to distinguish vagal or vasovagal syncope which is very common, the very stereotype reflex syncopes, carotid sinus hypersensitivity sometimes associated with sinus node dysfunction and borderline forms such as orthostatic sinus tachycardia and cerebrovascular syncope. The differential diagnosis is vast, from simple hysteria to severe cardiac disease. Tilt testing should be indicated for diagnosis of most cases of syncope with apparently normal hearts. Therapeutic abstention is the rule, providing certain preventive measures are taken, but, should treatment be necessary, cardiac pacing remains an exceptional modality in vasovagal syncope. Strict clinical and physiopathological studies are still required to determine the long-term prognosis and the underlying mechanisms of these syndromes. PMID- 9749284 TI - [Gaps: practical implications]. AB - The French translations of the word "gap" include "hole, breach, opening". In the Anglo-saxon literature, the conduction gap or the gap phenomenon is defined as the period of the cardiac cycle during which a premature beat is blocked whereas more delayed or more premature beats are conducted. This rare phenomenon, observed mainly during extrasystolic stimulation, was previously called supernormal conduction. The underlying mechanism is distal block in the conduction system occurring when more premature beats lead to delay in the proximal conduction which gives the distal site time to become excitable again. The excitability gap is one of the main characteristics of reentrant arrhythmias. Its presence in a reentrant circuit ensures the regularity and stability of the arrhythmia. It also allows penetration of the circuit by external stimuli. This different extrasystolic stimulation technique (resetting) or rapid fixed rate pacing (entrainment) may be used diagnostically and therapeutically. It enables the identification of the arrhythmia circuit and a critical zone (protected isthmus, zone of slow conduction) which may constitute a target for ablation. It also offers a possibility for terminating the arrhythmia by external stimulation. Finally, the duration of the excitable gap may guide the choice of antiarrhythmic agent during pharmacological cardioversion of a reentrant tachycardia. PMID- 9749285 TI - [Present concepts of accessory pathways]. AB - The possibility of ablation of arrhythmias has revived interest in accessory atrioventricular pathways. The authors propose a classification based on their electrophysiological properties, the nature of the arrhythmias which they induce, their anatomy and localisation. Classical accessory pathways have a rapid "all or nothing" type of conduction and are responsible for the WPW syndrome. They are called bundles of Kent although Kent's description does not correspond exactly to our present concept of their structures. However, some classical accessory pathways do have unidirectional conduction properties. When only retrograde reciprocating orthodromic tachycardia may occur but not preexcitation is observed in sinus rhythm: they have to be differentiated from reciprocating nodal tachycardias. Atypical accessory pathways show decremential conduction and are composed of specific tissues. When the conclusion is mainly or exclusively anterograde, tachycardias may be observed which were generally attributed to nodoventricular Mahaim fibres. When the conduction is essentially retrograde, they usually give rise to chronic junctional tachycardias. PMID- 9749286 TI - [Rhythmic pitfalls in cardiac pacing]. AB - Some electrocardiographic appearances in cardiac pacing may suggest pacemaker dysfunction but in fact the unit may be functioning normally or have a minor fault which is easy to correct by reprogramming. A pacing rate different to that programmed may be due to the rate-response hysteresis or rate smoothing functions. Irregular pacing is often due to phenomena of inappropriate sensing. A pacemaker in the bipolar AAI mode may seem to have no output if the spike is not visible: function in AAI mode should not be interpreted as pacing catheter displacement. The practician may wrongly interpret faulty ventricular sensing in patients with AAI pacemakers and atrial fibrillation when irregular pacing is observed or when ventricular extrasystoles do not inhibit the pacemaker. In dual chamber pacing, the blanking period may result in inadequate ventricular stimulation. The phenomenon of crossed detection or cross-talk is a cause of inappropriate inhibition. Applications of the magnet blocks the sensing function: the magnet pacemaker rate is an indicator of pacemaker end of life. In fact, the magnet induces different behaviours depending on the model of pacemaker which makes it essential to know the special characteristics of each pacemaker. The application of the magnet may trigger arrhythmias or no output of pacemakers at the end of life. The A-V interval may vary with respect to its response to the heart rate, when there is a hysteresis function of the A-V interval, sensing in the "safety" gap or when anti-atrial trachycardia algorithms are activated. The acceleration of a dual-chamber pacemaker may be related to electronic reentrant tachycardia or to an atrial tachycardia: a fault or delayed activation of various anti-arrhythmic algorithms may also cause difficulties in the interpretation of the electrocardiogram. PMID- 9749287 TI - [Recommendations of the French Society of Cardiology for the training of echocardiographers and performing echocardiograms]. AB - The considerable advances achieved in the field of echocardiography have made this investigation an essential diagnostic tool. Under the auspices of the French Society of Cardiology, the Working group on Echocardiography publishes its practical recommendations for optimising the training of echocardiographers (theoretical instruction and practical courses) and for performing echocardiography (understanding the clinical problem, referral to previous examinations, necessary recordings and measurements, and appropriate equipment). In the future, these recommendations should be updated to take into account continuing technical improvements and changes in methods of studying cardiac disease by echocardiography. PMID- 9749288 TI - [Recommendations on the practice of intracavitary diagnostic methods and treatment of cardiac arrhythmias. Diagnostic electrophysiology; interventional electrophysiology; permanent cardiac stimulation; automatic implantable defibrillators]. PMID- 9749289 TI - [Recommendations of the French Society of Cardiology concerning the training of specialists in coronarography and angioplasty, organization of equipment for centers for coronarography and coronary angiography]. PMID- 9749290 TI - [Recommendations of the French Society of Cardiology concerning the practice of exercise tests in adult heart patients]. PMID- 9749291 TI - [Recommendation of the French Society of Cardiology concerning the practice of cardiovascular rehabilitation in adults]. PMID- 9749292 TI - [Recommendations of the French Society of Cardiology concerning the indications and surveillance of oral anticoagulant therapy]. PMID- 9749293 TI - [Recommendations of the French Society of Cardiology on the question of including adults, excepting emergencies, in waiting lists for cardiac transplantation]. PMID- 9749294 TI - [Shoulder complaints in family practice; a simple approach]. AB - In two patients with shoulder complaints, a man aged 56 years and a woman aged 43, the function of the shoulder girdle (structures of the cervical spine, the upper thoracic spine and the upper ribs) played an important role. A simple classification of shoulder complaints is presented in which the influence of the shoulder girdle is recognized: (a) synovial disorder: complaints originating from the structures of the glenohumeral joint, the subacromial space, the acromioclavicular joint or combinations of these, (b) shoulder girdle disorder: the complaints are caused by functional problems in the structures of the shoulder girdle, (c) combination disorder: both a synovial disorder and a functional disorder of the structures of the shoulder girdle are causing the complaints, making it impossible to determine which structure is the primary cause. Considering the limited therapeutic choices (at first nonsteroidal antiinflammatory drugs, followed in case of persistent complaints by injections of analgesics and corticosteroids for synovial complaints, or manipulative therapy for shoulder girdle complaints), a more detailed classification is not needed to determine a successful therapeutic strategy. PMID- 9749295 TI - [Proteins with frameshift mutations in Alzheimer's disease; cause or effect?]. AB - In a recent paper in Science, a novel finding was presented on Alzheimer's disease. The authors describe mutant proteins, brought about by a change in the mRNA reading frame. These so-called +I proteins appear to be associated with neuropathological changes in Alzheimer's disease brains. Two proteins involved in the pathogenesis of the disease were studied: beta-amyloid precursor protein en ubiquitin-B. For both proteins, a deletion of two bases in a small percentage of the mRNAs caused the frame shift. Possible mechanisms and implications are discussed. PMID- 9749296 TI - [Presenilins as a marker of Alzheimer's disease]. AB - Although the sporadic form of Alzheimer's disease is the most common, rare familial variants exist. Approximately 50% of these cases are caused by a mutation in the presenilin genes. Mutations in presinilin genes give rise to Alzheimer's disease in a dominant pattern of inheritance with an early age of onset (< 60 years). Both presenilins (PS-1 and PS-2) are transmembrane proteins localized in the intracellular membranes of the endoplasmatic reticulum and Golgi apparatus. This suggests they play a role in transport or sorting of proteins in the cell. Different lines of evidence directly link presenilin to the formation of beta-amyloid, an important constituent of senile plaques. PS-1 has an essential function during development: mice lacking intact PS-1 are not viable. In addition, structural and functional homologies have been identified between presenilins and Notch signal transduction pathways, which play a role in development. The discovery of the presenilin mutations has provided a new angle to Alzheimer's disease research. Eventually, this will probably greatly contribute to knowledge of the pathogenesis of the disease and in time support the development of novel therapeutic strategies. PMID- 9749297 TI - [Influenza A (H5N1) in Hong Kong: Forerunner of a pandemic or just a scientifically interesting phenomenon and a useful exercise in pandemiology?]. AB - In 1997, 18 influenza patients were detected who were infected with influenza A(H5N1) virus. Six patients died. Presumably most of the patients had acquired the infection directly from chickens with the fowl plague prevalent in China in 1997. These are the first reported cases of isolation of influenza viruses belonging to one of the H4-H15 subtypes from human influenza patients. Man-to-man transmission of the virus has not been demonstrated but cannot be excluded in every case. Genetic analyses of seven of these virus isolates showed that no reassortment with a human or porcine influenza virus had occurred. It is unpredictable whether the H5N1-virus in question will start a pandemic in the next few years. PMID- 9749298 TI - [Prosthesis of the upper extremity]. AB - In functional impairment of a joint due to pain and (or) limited mobility in which conservative measures are inadequate, an operation may be considered. The principal possibilities are arthrodesis and implantation of an artificial joint. Reasons to opt for a prosthesis are involvement of adjacent articulations (in rheumatoid arthritis) and preservation of mobility which, together with stability, is necessary for good articular function. The principal indications for implantation of an artificial joint in the upper extremity are rheumatoid arthritis and (to a lesser degree) osteoarthritis and posttraumatic impairments. The functional results of prostheses in the shoulder, elbow and wrist are good, but the proportion of complications is relatively large (compared with that in hip and knee prostheses) and follow-up so far has only been short. PMID- 9749299 TI - [Results of anterior shoulder decompression surgery according to Neer for shoulder impingement syndrome; little effect on fitness for work]. AB - OBJECTIVE: To evaluate the effect of surgical treatment of the impingement syndrome of the shoulder on the fitness for work. DESIGN: Retrospective. SETTING: Academic Hospital Groningen, department of Orthopaedics, the Netherlands. PATIENTS AND METHODS: A group of 31 patients who had undergone an anterior shoulder decompression between 1 January 1984 and 31 December 1993 because of shoulder symptoms due to impingement lasting more than one year, were subjected in January 1996 to a study of the effect of the treatment on the pain, fitness for work and participation in the labour process. The results were measured using an objectivated score list for pain and function and with a comparison of the labour situations before and after the operation as described by the patients. RESULTS: End results as measured with the score list were fair to good in 97 patients (74%). The numbers of those completely fit for work before operation and at follow-up were identical, viz. 45 out of 131 (34%). Thirty-four patients (26%) had other jobs or adjusted activities and 37 (28%) were completely unfit for work (together 71 patients: 54%); 15 patients (11%) had stopped working for reasons of age. The probability to be definitely rejected for work after the operation was increased significantly by a job causing shoulder overstrain and by having been on sick leave with pay prior to the operation. CONCLUSION: Although the objective results were reasonably good, the percentage of those completely fit for work was not changed appreciably by the surgical treatment. PMID- 9749300 TI - [Leptospirosis in the Netherlands, 1991-1995]. AB - OBJECTIVE: To determine all cases of leptospirosis in the Netherlands in 1991 1995 that were confirmed by serological investigation or culturing. DESIGN: Descriptive, retrospective. SETTING: Department of Biomedical Research, Royal Tropical Institute, Amsterdam, the Netherlands, and Department of Internal Medicine, Academic Medical Centre, Amsterdam, the Netherlands. METHOD: Using data of the Reference Laboratory for Leptospirosis, the number of leptospirosis cases in 1991-1995 was determined and compared with the data of 1986-1990. Additional information was obtained about patients with confirmed leptospirosis or who had been hospitalized because of leptospirosis. RESULTS: The number of confirmed cases dropped from 229 in 1986-1990 to 159 in 1991-1995. This decrease could be attributed mainly to a marked decrease of the number of dairy farm fever (hardjo) cases. There was a clear increase of the number of infections acquired during travel in foreign countries, notably outside Europe. Thus, leptospirosis in the Netherlands shifted from an occupational disease towards a disease due to recreational activities. In about 10% of the patients the disease ran a severe course with Weil's syndrome (icterus, renal failure, and haemorrhages). Eight patients (5%) died. Clinical data of 5 of these 8 patients indicated that they had suffered from Weil's syndrome. PMID- 9749301 TI - [Integrated (biopsychosocial) treatment in the family practice: a pregnant patient with a sacral tumor and severe posttraumatic stress disorder]. AB - In a 25-year-old woman pregnant for the second time after a successful first pregnancy, a locally aggressive, invasive sacrum tumour was diagnosed. The execution of the necessary but potentially mutilating surgical procedures was seriously hampered even during the preparative phase, in spite of the conscious wish of the patient to comply, by her severe psychiatric problems (posttraumatic stress disorder with dissociative symptoms). The Psychiatric Consultation Service took over the case management and an integrated (biopsychosocial) diagnostic investigation was carried out, involving analysis of the problems on four system levels: the biological, the psychological, the social and the health care level. An integrated treatment plan was drafted. By collaboration of the entire multidisciplinary treatment team conditions were secured under which patient would let herself be treated. In this way she was enabled to undergo the necessary procedures, with good results. PMID- 9749302 TI - ['Public health status and perspectives' 1997. IV. Health status and life expectancy weighted]. AB - The construction of composite measures of population health status meets the need to combine data on mortality and morbidity into one single population health index. Such indices can serve in principle to monitor population health status over time, or as support for the allocation of resources. In the framework of the 'Dutch public health status and forecasts report' for 1997 several calculations, new for the Netherlands, are made along the lines of both the 'health expectancy and the 'disability-adjusted life years' (DALY) concepts. PMID- 9749303 TI - ['Public health status and perspectives' 1997. V. Effects of prevention]. AB - For the 'Public health status and forecasts' 1997 attention has been given to the health benefit that could still be achieved by primary and secondary prevention. For selected examples of five types of methods of prevention (health education, vaccination, legislation and regulation, pharmaceutical prophylaxis and screening) the efficacy (the health benefit which can be achieved under optimal circumstances) is compared with the effectiveness (the health benefit achieved in practice). The potential health benefit through interventions aimed at (life style related) determinants is considerable. It is advised to work towards a programmatic prevention strategy giving attention to ascertaining the correct target group and the most suitable intervention strategy, to regular evaluation of the effectiveness, to implementing an intersectoral approach and to clearly establishing the responsibilities at both the central and the local level. PMID- 9749304 TI - [Psychiatric disturbances in asylum seekers and refugees: is psychological trauma the cause?]. PMID- 9749305 TI - [Psychiatric disturbances in asylum seekers and refugees: originating from psychological trauma?]. PMID- 9749306 TI - [Impression of the thighs; lipoatrophia semicircularis]. PMID- 9749307 TI - [Hormonal treatment of constitutionally tall children]. PMID- 9749308 TI - [Painful breasts]. PMID- 9749309 TI - [Painful breasts]. PMID- 9749310 TI - [Polysaccharides of flower plants: structure and physiological activity]. AB - The results of studies on the chemical structure and physiological activity of phanerogam polysaccharides, accumulated within the last two decades, are reviewed. Three types of polysaccharides are considered: rhamnogalacturonans (pectins and related gums and mucilages, type A), acidic arabinogalactans (mainly plant mucilages, gums, and some hemicelluloses, type B), and neutral glucans and heteroglycans (reserve polysaccharides, type C). Various physiological activities of these plant polysaccharides are discussed, with particular emphasis being placed on their immunomodulatory action. The data available on the relationship between chemical structure and physiological activity of plant polysaccharides are considered. Information on the medicinal use of some plants containing physiologically active polysaccharides is presented. PMID- 9749311 TI - [Synthesis and biological properties of artificial antigens based on 280-289 fragment of beta2-gpI]. AB - A series of artificial antigens were synthesized on the basis of the FC(Acm)KNKEKKC(Acm)S peptide from the beta 2-glycoprotein I sequence: lipophilic analogues, the peptide-BSA conjugate, and multiple antigen peptide (MAP) containing eight copies of the peptide on an oligolysyl core. The solid phase method for acylation of the peptide with fatty acids and the HPLC analysis of the acylpeptides were described. Antigenic properties of the resulting compounds were evaluated by CL-ELISA. PMID- 9749313 TI - [Architecture of compact three alpha-helical structures]. AB - Modeling and methodical analysis of possible compact spatial arrangements of three alpha helices bound with connections were carried out. It was suggested to describe the compact three-alpha-helical structures as combinations of alpha helical hairpins, L-shaped structures, V-shaped structures, alpha-alpha comers, and alpha-l-alpha motifs. Practically all the structures resulting from such a modeling were shown to exist in globular proteins. Many small proteins and domains were found to consist of only these three-helical structures. This indicated that each such three-helical structure was stable by itself and capable of independent folding of its polypeptide chain into this structure. PMID- 9749312 TI - [Low-molecular cytolysins and trypsin inhibitors from sea anemone Radianthus macrodactylus. Isolation and partial characterization]. AB - Two low-molecular cytolytic toxins (RmI and RmII) and four trypsin inhibitors were isolated from the aqueous extract of sea anemone Radianthus macrodactylus. The method of isolation involved precipitation with acetone, gel filtration on acrylex P-4, ion-exchange chromatography on CM-32 cellulose, affinity chromatography on trypsin-binding sepharose 4B, ion exchange chromatography on an Ultrapore TSK CM-3SW column, and reversed phase HPLC on a Silasorb C18 column. RmI, RmII, and JnI inhibitor displayed molecular masses 5100, 6100, and 7100 Da, respectively, when subjected to SDS-PAGE. The isoelectric points were 9.2 and 9.3 for RmI and RmII, respectively. The amino acid composition and N-terminal amino acid residue (glycine) were determined for RmI, RmII, and JnI. Both proteins were nontoxic to mice and crabs. Hemolytic activity was determined to be 25 and 20 HU/mg for RmI and RmII, respectively, and their action on erythrocyte membrane was not inhibited by exogenous sphingomyelin. RmI and RmII exhibited antihistamine activity. PMID- 9749314 TI - [Production of recombinant human keratinocyte growth factor in Escherichia coli cells]. AB - Here we describe the synthesis of the gene encoding human keratinocyte growth factor (hKGF) and the construction of vectors for cytoplasmic production of hKGF in Escherichia coli cells. The level of recombinant protein expression was 1-1.5% of the total cell protein. hKGF was purified to homogeneity by three-step ion exchange and affinity chromatography. The protein is biologically active: it stimulates dose-dependent protein synthesis in cultured human epidermal keratinocytes. PMID- 9749315 TI - [Regulation of synthesis of ribosomal protein L7-L12: role of intercistronic rplJL region as an enhancer of translation]. AB - The ability of the Escherichia coli intercistronic rplJL region to initiate effectively the synthesis of the ribosomal protein L7/12, the only ribosomal component present in the ribosome in four copies rather than in one was studied in vivo and in vitro. It was shown that the structural determinants located upstream from the Shine-Dalgarno sequence and sharing structural motifs with the known E. coli translational enhancers are necessary for high activity of this region in translation initiation. These data indicate that mRNA-protein interactions through the ribosomal S1 protein play an important role in the formation of the initiation complex, and an enhancer region within the leader of the L7/12 mRNA serves as a target for this protein. PMID- 9749316 TI - [Stabilized reaction mixture for in vitro mRNA translation]. AB - Here we describe a method for obtaining a ready-to-use stabilized reaction mixture for in vitro translation of mRNA. We also demonstrate the stabilization of a complete translation mixture containing wheat germ extract, amino acids, ATP, GTP, creatine phosphate, creatine kinase, and the reaction buffer by lyophilization in the presence of various sugars. The greatest stabilizing effect is achieved by supplementing the mixture with 10% (mass/volume) trehalose, which is also a unique translation activator, enhancing the translation of various mRNAs. A lyophilized complete translation mixture containing trehalose can be stored at 4-8 degrees C for several months without losing its activity. The mixture can be easily reconstituted by adding an aqueous mRNA solution and retains the potential for reproducible functioning. This allows the employment of such a cell-free translation system for analytical screening of a broad spectrum of compounds inhibiting translation at various stages. PMID- 9749317 TI - [Inactivation and stabilization of urokinase fibrinolytic activity in vitro]. AB - When incubated at 30 degrees C and pH 7.4, urokinase lost fibrinolytic activity (i.e., the plasminogen-activating activity measured by the time of fibrin clot lysis) but completely retained amidase activity. The enzyme inactivation rate depended on the urokinase concentration and, at concentrations of more than 1.5 microM, was described by a second order equation, which indicated that the enzyme underwent autolytic degradation (kaut = 3.8 x 10(-3) M-1 min-1). During incubation, urokinase (54 kDa) was converted into its low-molecular-mass form (33 kDa) and products of the A-chain degradation. The amidase activity did not correlate with the fibrinolytic activity in the cases when the enzyme molecule underwent local unfolding or partial degradation. The optimum mixture of agents for stabilizing the fibrinolytic activity of urokinase was found. PMID- 9749318 TI - [New method of isolation of O-specific polysaccharide from gram-negative bacteria Yersinia pseudotuberculosis]. AB - Using bacteria Yersinia pseudotuberculosis as an example, two procedures for obtaining the O-specific polysaccharide of endotoxins of gram-negative bacteria were compared: the direct acidic hydrolysis of whole cells and the traditional procedure based on preliminary isolation of the corresponding lipopolysaccharide. Analysis of the resulting polysaccharides showed that the isolation from the bacterial biomass gave a high yield of the polysaccharide; the polysaccharide is not degraded; and judging from the 13C NMR data, its structure was completely identical to that of the corresponding product of lipopolysaccharide hydrolysis. Therefore, this procedure is useful for obtaining O-specific polysaccharides for structural studies. PMID- 9749319 TI - Beyond single mothers: cohabitation and marriage in the AFDC program. AB - We investigate the extent and implications of cohabitation and marriage among U.S. welfare recipients. An analysis of four data sets (the Current Population Survey, the National Survey of Families and Households, the Panel Study of Income Dynamics, and the National Longitudinal Survey of Youth) shows significant numbers of cohabitors among recipients of AFDC. An even more surprising finding is the large number of married women on welfare. We also report the results of a telephone survey of state AFDC agencies conducted to determine state rules governing cohabitation and marriage. The survey results indicate that, in a number of respects, AFDC rules encourage cohabitation. Finally, we conduct an analysis of the impact of AFDC rules on cohabitation, marriage, and single motherhood and find weak evidence in support of incentives to cohabit. PMID- 9749320 TI - Changes in assortative mating: the impact of age and education, 1970-1990. AB - Data from the U.S. Census and Current Population Survey are used to examine trends in the propensity to marry or to cohabit by the age and educational attainment of potential partners. Marriage rates declined sharply across all age and educational combinations between 1970 and 1980 and declined more sharply for less-educated persons between 1980 and 1990. The rise in cohabitation compensated somewhat for the decline in marriage rates, but the compensation was unequally spread among age and educational combinations. Highly educated men were more likely, and highly educated women were no more or less likely, to marry than to co-habit with less-educated partners in 1970 and 1980. By 1990, however, educational assortative-mating patterns between these two types of unions were similar. In 1990, marriages and cohabitations involving women who were better educated than their partners were more common than those involving women who were less educated than their partners. In addition, men and women in their early 20s tend to have partners better educated than themselves, but persons in their 30s tend to cross the less-than-high-school/more-than-high-school educational barrier when partners differ in educational attainment. PMID- 9749321 TI - Family and sociodemographic influences on patterns of leaving home in postwar Britain. AB - We identify child-level and parent-level characteristics associated with children's patterns of leaving home. We use a multilevel discrete-time hazards model to examine the impact of family and demographic factors at both levels, and utilize the Alternating Conditional Expectation algorithm optimally to transform the dependent and independent variables. We find that measured variables at both the child and the parent level have important influences, as do period and cohort factors. However, unmeasured parent-level factors have an influence on the departure of children that is broadly similar in magnitude to measured factors. PMID- 9749322 TI - Reassessing the decline in parent-child old-age coresidence during the twentieth century. AB - The share of the elderly living with an adult child decreased monotonically throughout the twentieth century, while the probability of reaching old age and the number of years lived in old age increased. As a result, the expected number of life-years lived with adult children while in old age may have increased, decreased, or stayed the same. I estimate that the number of life-years lived in old-age coresidence with adult children stayed roughly constant between 1900 and 1940, while the rate of coresidence declined. Life-years lived in old-age coresidence then declined substantially between 1940 and 1990. Moreover, the number of life-years lived in old-age coresidence in 1990 would have been roughly half as great as it actually was had there been no improvements in mortality between 1900 and 1990. And if fertility had remained at its 1900 levels, life years lived in old-age coresidence would have been about 45% higher in 1990 than it actually was. The results imply that analyses of the change in familial assistance to the elderly should also consider changes in mortality. PMID- 9749324 TI - Ethnic stratification in northwest China: occupational differences between Han Chinese and national minorities in Xinjiang, 1982-1990. AB - The debate on market reforms and social stratification in China has paid very little attention to China's ethnic minorities. We explored rising occupational stratification by ethnicity in the Xinjiang Uygur Autonomous Region. Analyses of census data from 1982 and 1990 pointed to educational disadvantages faced by ethinic minorities as the most plausible explanation for the change. Multivariate analysis revealed a significant increase in the effect of education on high status occupational attainment but no change in the effect of ethnicity. Net of education, ethnic differences in high-status occupational attainment were negligible. In contrast, large ethnic differences in manufacturing and agricultural occupations persisted after education and geography were statistically controlled. PMID- 9749323 TI - Couple childbearing plans and births in Sweden. AB - We use data from a nationally representative sample of Swedish couples to estimate effects of partners' childbearing plans on the rate of subsequent childbearing. Only 11% of the couples in this sample expressed plans in opposite directions (plan to have a child versus not to have a child), but 24% had differing levels of certainty about their plans. Of the couples in which both partners said they definitely planned to have another child, 44% had a child within two years. If neither partner planned to have another child, less than 2% of couples had a birth. The figure was 6% if the partners had opposing childbearing plans. Thus, both men and women exerted veto power over further childbearing. Disagreements were equally likely to be resolved in favor of the woman as of the man, and effects of partners' plans on the birth hazard did not depend on the couple's gender arrangements, family ideologies, or marital status. We discuss these results in the context of Sweden's public support for gender equality and for childrearing, its pervasive contraceptive regime, and its high rates of cohabitation. We also argue for the collection of data from partners in future family and fertility surveys. PMID- 9749325 TI - Gender, migration, and career trajectories in Malaysia. AB - With data from the Malaysian Family Life Survey, I use a continuous-state hazards model to study the impact of migration on the dynamics of individuals' careers. I distinguish between the effects of family migration and solo migration by gender. The results show that migration alters the career trajectory primarily by accelerating the process of occupational mobility rather than by increasing the level of occupational attainment. Further, the effect of migration on careers varies by type of migration, especially for women. Male-female differences in the outcome of family migration, however, are visible only in transitions into and out of employment. PMID- 9749326 TI - Mortality in Vietnam, 1979-1989. AB - Little is known about past and present mortality in Vietnam, as the first official data on mortality have only recently become available from censuses taken in 1979 and 1989. Using these data, I estimate Vietnamese mortality during the intercensal period using two techniques that rely on age-specific growth rates from two successive age distributions. Intercensal emigration and differential completeness of census enumeration associated with massive outflows of refugees in the wake of the Vietnam War, population-redistribution policies, and a highly mobile population represent important sources of bias for the estimation of intercensal mortality. I incorporate several strategies to minimize bias from these sources and to select the method that is least sensitive to errors associated with them. Life expectancy at birth estimated for the 1979-1989 intercensal period is 61.4 years for males and 63.2 for females. These results suggest a trend of declining mortality between the 1970s and the 1980s and add solid empirical evidence to the debate over whether mortality in Vietnam has been deteriorating or improving. PMID- 9749327 TI - Educational attainment and transitions in functional status among older Taiwanese. AB - Despite considerable research examining the influence of socioeconomic status on health, few studies have considered this relationship as it pertains to older adults in non-Western societies. We attempt to ascertain the influence of education on changes in physical functioning in a rapidly developing country. Data come from the 1989 Survey of Health and Living Status of the Elderly in Taiwan and a follow-up interview in 1993 (N = 4,049, age = 60+). Individuals are conceptualized to be in a state of functional independence or functional limitation at the time of origin, based on their ability to perform three physical functioning tasks. The outcome at the follow-up interview is categorized as functionally independent, limited, or dead, allowing for six probabilities, one from each state of origin to each outcome. These are calculated using a multinomial logit model, controlling for other factors often thought to be associated with health transitions. High levels of educational attainment result in a decreased incidence of functional limitation for those originating in a state of independence. Contrary to expectations, however, education has little influence on those who originate functionally limited. Thus, higher education plays a substantial role in primary prevention of morbidity, delaying the onset of disability, but other factors are more important once limitations begin. We speculate on the reasons behind these findings, including that the results may be culturally dependent. PMID- 9749328 TI - [Expression of the human dystrophin gene in mdx mouse muscle fibers after transfection using liposomes and synthetic oligopeptides]. AB - The number of dysrophin-positive fibers appearing in the femoral quadriceps muscle of mdx mice after injection of the full-length human dystrophin cDNA within the pHSADy plasmid was examined by means of immunohystochemical techniques. Transfection was carried out using lipofectamine (LFA), or synthetic oligopeptide complexes that provided the condensation of plasmid DNA (K8) and its release from endosomes gopeptide complexes that provided the condensation of plasmid DNA (K8) and its release from endosomes (JTS1). The LFA + pHSADy at a dose of 10 micrograms DNA did not affect the number of dystrophin-positive fibers at the site of injection (0.6-0.8%), whereas it caused a statistically significant increase in the number of these fibers in the same muscle of the contralateral leg (up to 2.3%). Injection of the SO + pHSADy complex resulted in the occurrence of dystrophin-positive muscle fibers characterized by a heterogeneous content and the distribution of dystrophin. The greatest number of dystrophin-positive fibers (about 16%) was observed under a ratio of pHSADy to K8 of 1:3 or 1:4. The observed maximal number of dystrophin-positive fibers after a single injection of SO + pHSADy was 3.8%, and it was 17.7% after three injections. These values were statistically significantly higher compared to intact mice (0.6%), the injection of pure plasmid (2.2%), or the intramuscular injection of sucrose (from 0.7 to 1.3%). A relatively high level of transfection (about 5%) was observed after an intracardiac injection of a large dose of the pHSADy (70 micrograms DNA). The perspectives of the targeted delivery of the dystrophin gene into muscles under conditions of parenteral administration are discussed. PMID- 9749329 TI - [Molecular phylogeny of the nuclear 5.8S ribosomal RNA genes in 37 species of Nicotiana genus]. AB - The primary structure of the 5.8S rRNA gene was established in 37 species of the genus Nicotiana representing four different sections of the subgenus Petuniodes: Acuminatae, Alatae, Noctiflorae, and Suaveolentes. On the basis of these data, phylogenetic analysis was performed. The obtained results are in good agreement with the modern systematics of the genus. However, the bootstrap analysis gives only poor statistical support for the branches of the phylogenetic tree owing to a small number of phylogenetically informative positions. PMID- 9749330 TI - [Different patterns of molecular evolution of influenza A viruses in avian and human population]. AB - Patterns of molecular evolution of the influenza virus proteins and genes are discussed. The subsets of all viral genes corresponding to statistically significant clusters on dendrogram were shown to fall into two distinct groups. The first group was characterized by the presence of an exact linear relationship between the year of the strain isolation and the evolutionary distance. The subsets of human influenza virus genes belong to this group. A method for eliminating the "frozen" strains from the subsets and for calculating the evolutionary rates without construction of phylogenetic trees has been elaborated. The substitution rates calculated according to this technique agreed with the data obtained previously. A linear relationship was not observed in the second group. This group was predominantly composed of avian influenza virus genes. The lack of linear correlation pointed to the cocirculation of a large amount of different influenza virus genomic segments in the avian population. An approach for an examination of the role of intragenic recombination in the development of the antigenic subtypes of hemagglutinin is suggested. Our results suggest that recombination did not play a considerable role in this process, and that all modern subtypes of this protein were probably formed before the introduction of the influenza viruses into the human population. These findings are consistent with the hypothesis that influenza viruses penetrated into human population from their pools in avian populations. PMID- 9749331 TI - [Lethal and mutagenic effects of tritium incorporated into position 8 of the purines in phage lambda DNA and the role of the Fpg protein]. AB - The lethal and mutagenic effects of the decay of 3H incorporated in phage lambda DNA as 8-3H-adenosine and 8-3H-guanosine were studied, using the DNA of 8-3H adenine as a labeled DNA precursor. A transmutation component of 3H decay is involved in formation of 8-oxoguanine (8-oxo-G) and 8-oxoadenine (8-oxo-A) residues in phage DNA. The efficiency of phage inactivation (the number of lethal lesions per one tritium decay in the phage genome) for 3H decay in position 8 of purines was the same as that measured in positions 5 and 6 of pyrimidines (alpha = 0.14 +/- 0.01) and virtually did not depend on the fpg-1::kan mutation in the host gene encoding the Fpg protein (formamidepyrimidine-DNA-glycosylase). The efficiency of the mutagenic effect of 3H-purines Em (frequency of c mutations per one 3H decay in the phage genome) was (2.9 +/- 0.3) x 10(-5) in the fpg+ host and (4.6 +/- 0.4) x 10(-5) in the fpg-host. This means that the Fpg protein excised approximately 40% of premutational DNA lesions (probably, 8-oxo-G residues). Induction of the mutagenic SOS system by UV light caused a 1.5-fold increase in the frequency of c mutations induced by 8-3H-purines in fpg+ cells over that in fpg-cells. This suggests that apurinic AP sites produced after the excision of 8 oxo-G by the Fpg protein are substrates for mutagenic SOS repair. PMID- 9749332 TI - [Allozyme heterozygosity, sexual maturation rate, and longevity]. AB - Humans, animals, and plants are shown to have a universal polygenic system maintained by stabilizing selection, which underlies highly significant correlations between multilocus allozyme heterozygosity, the sexual maturation rate, and longevity. For 77 animal and 30 plant species, it was demonstrated that allozyme genomic heterozygosity (1) is positively correlated with rate of sexual maturity (2) (r12 = 0.815, P < 0.001) and negatively correlated with longevity; (3) (r13 = -0.793, P < 0.001). These relationships are significantly affected by the genotype-environment interactions that modify the structure of the metabolism related to growth and development. An increase in the number of heterotic genes involved in growth and development increases the energy expenditure in this ontogenetic period, accelerates the sexual maturation rate, and shortens the life span. PMID- 9749333 TI - [Development of thermotolerance in Drosophila melanogaster line l(1) ts403 with a defect in heat shock protein synthesis]. AB - To elucidate the role of heat shock proteins (HSP) in the formation of resistance to extreme factors and in the development of organismic and cell response to these factors, thermotolerance in a Drosophila melanogaster line with defective HSP synthesis was studied with regard to several criteria: (1) survival of adult females; (2) damage to egg chambers in ovarioles; (3) dynamics of oviposition; (4) frequency of loss and nondisjunction of sex chromosomes in meiosis of females exposed to a heat shock (HS). According to all these criteria, the l(l)ts403 females were more sensitive to a HS 37 degrees C, the exposure at 37 degrees C for 1 h (HS37) than the females of the wild-type line Canton S. Only the data on the first three aforementioned parameters were indicative of thermotolerance development. In files exposed to HS35 followed by HS37, a decrease in nondisjunction and loss of sex chromosomes in the 3- and 4-day ovipositions was observed as compared to the flies exposed to HS37 only. This can be explained by the differences in oviposition dynamics and, consequently, in the realization rate of the oogenesis stages in these two experimental variants. The pleiotropic effect of the l(l)ts403 mutation that led to a disturbance in HSP gene expression at the posttranscriptional level and to an increase in the frequency of sex chromosome nondisjunction in the meiosis of females exposed to HS37 suggests that these processes were connected. As no thermotolerance was revealed by studying the sex chromosome nondisjunction, but thermotolerance was found by estimating the other parameters, we suggest that the product of the gene studied is involved in a signal system operating at a stage that precedes the HS-induced changes in the translation and division apparatuses of a cell. PMID- 9749334 TI - [Population dynamics of the response of the genomic pattern of the mobile genetic element Dm412 in Drosophila on selection for a quantitative trait]. AB - In an isogenic line of Drosophila melanogaster carrying the Mendelian mutation radius incompletus, selection for the total length of two segments of the disrupted longitudinal wing vein was conducted. After gamma-irradiation at a dose of 13 Gy, positive and negative truncation selection became highly effective and was completed in 50 generations. The pattern of mobile genetic element Dm412 was almost completely fixed in the course of selection. In the positive direction of selection, fixations of mobile genetic element (MGE) sites exceeded losses; in the negative direction, this relationship was reversed. The number of MGE sites in the pattern increased from 23 to 33 and to 26 in the positive and negative directions, respectively. The mean heterozygosity of MGE sites decreased respectively ten and six times. The dynamics of some sites (6F, 43B, 66A, 69E, and others) corresponded to that expected with an adaptive response to selection. Two out of these sites (43B and 66A) were previously assigned to hot sites of Dm412 transposition induced by heat shock. Fixation and loss of sites continued on average for tens of generations. Four hypotheses describe the relationship between patterns of polygenes and MGE in the context of explanation of the above facts: (1) genetic drift; (2) the linkage of MGE and polygenes without modification of the latter (hitchhiking); (3) the linkage and modifying effect of MGE on polygenes linked with them; (4) the selection of the "champion" pattern of polygenes and a random or adaptive MGE pattern linked with it. Hypotheses 1 and 2 are unlikely, hypothesis 3 is possible in the case of other selection modes, whereas hypothesis 4 seems to be most plausible. PMID- 9749335 TI - [Biosynthesis of testosterone in the gonads in silver fox embryos after long-term selection for domesticated behavior]. AB - Fetal gonad weight and testosterone content in serum and gonads were analyzed in silver fox every five days from the 35th day of pregnancy until delivery. Fetal testicles were also tested for testosterone production induced by chorionic gonadotropin (CG) in vitro. Pregnant females were sampled from an experimental population subjected to selection for domesticated behavior and a commercial population (control). Fetal gonad weight was significantly lower in domesticated animals than in controls. No differences were revealed in the testosterone contents in their serum and gonads and in the basal production of testosterone in fetal testicles. CG-induced production of testosterone was detectable from the 40th day of fetal development in domesticated animals and from the 50th day in controls. The results obtained suggest that domestication results in the hetero chronic fetal development of the hypophysial-testicular complex in silver fox. PMID- 9749336 TI - [Genotypic variability in populations of the mound-builder mouse Mus spicilegus Pet., 1882, at various life cycle stages]. AB - Genotypic variability in two Moldovian populations of moundbuilder mice Mus spicilegus Pet. was examined at different stages of their life cycle--at the beginning of overwintering in mounds and in the middle of summer in agrocenoses. Thirty allozymes were assayed. Significant differences in allele frequencies and heterozygosity between overwintered and mice born in the year of the experiment were recorded only for Idh-1. This fact was related to the presence of differential mortality among overwintered mice in the populations by the middle of summer. The increase in heterozygosity for Idh-1 in the populations occurred because of "old" mice that lived until the middle of summer, at which point they still have not contributed to the gene pool of the population. The genetic effect of the annual separation of generations of mound-builder mice due to their overwintering in mounds is analyzed. Overwintering has a bottleneck effect on the population. The relatively low genetic variability in mound-builder-mice populations is considered with regard to their biological and ecological features. Based on obtained data, a conclusion on the high genetic stability of moundbuilder-mice populations during their entire life cycle is made. PMID- 9749337 TI - [The ecologic component of the geographic variation of the Northern Eurasian gene pool and its archaeological age]. AB - The problem of the relationship between the state of the gene pool and population health was formulated. The subdivision into latitudinal zones, which is a characteristic feature of the northern Eurasian gene pool, was studied. This subdivision was expressed in the geographical distribution of the second principal component (PC2.G) of the total variance of 100 allelic frequencies for 34 polymorphic loci. It was suggested that this distribution of PC2.G resulted from the genetic adaptation of the population to the latitudinal zonality that is characteristic of northern Eurasia. It was also suggested that the latitudinal geographic variation was not inherent in the northern Eurasian gene pool and was younger than its total historical age. The evolutionary age of the principal components of the gene pool was estimated based on the geography of the principal components of the geographic variability of the late Paleolithic material culture, as well as the paleogeography of the Quaternary period. It was demonstrated that the first principal component of the modern gene pool was almost completely formed in the late Pleistocene; hence, its age is at least 16,000-26,000 years. The age of the second principal component (PC2.G) was estimated at less than 12,000-15,000 years; it was mostly formed in Holocene. A hypothesis was advanced that genetic adaptation occurs at the cost of population health, with the cost increasing as the evolutionary age of the genetic adaptation decreases. It was suggested that the latitudinal zonality of gene pool was accompanied by a similar zonality in the geography of morbidity as a cost of genetic adaptation. PMID- 9749339 TI - [Polymorphism of the HLA-DRB1 04 gene in inhabitants of Tuva]. AB - Data on the HLA-DRB1 04 gene typing among Tuvinians from Kyzyl are presented. Experiments were conducted by using the hybridization of allele-specific oligonucleotides with the amplified fragments of high molecular-weight DNA and reverse hybridization. The HLA-DRB1 0401 allele was most frequent among DR4 positive individuals. The HLA-DRB1 0405 and HLA-DRB1 0403 alleles were found at somewhat lower frequencies. In one individual, the rare HLA-DRB1 0410 subtype differing from HLA-DRB1 0405 subtype by the substitution of glycine for valine in codon 86 was detected. PMID- 9749338 TI - [Diversity of hereditary pathology in the population of Marii El Republic and its differentiation with respect to gene frequencies for hereditary diseases]. AB - The diversity of Mendelian hereditary pathology was studied in Marii El Republic. In total, 276,900 subjects, including 171,151 Maris and 88,714 Russians, living in seven raions (districts) were studied. Fifty-five autosomal dominant disease entities were found, with more than ten diseases having a frequency of 1:50,000 people or higher. In Maris, autosomal recessive hypotrichosis was observed at a relatively high frequency (1:15,337); this disease was not revealed in the Russian population studied earlier. Conversely, no phenylketonuria (PKU) was found in Maris, while it was a relatively common autosomal recessive disease in Russians. Regarding autosomal dominant pathology, 76 disease entities were revealed, with 21 diseases being observed at a frequency of at least 1:50,000. Ten X-linked diseases were found. The numbers of both autosomal recessive and autosomal dominant diseases exhibited a linear relationship with the number of subjects examined. The genetic structure of the Mari population was studied on the basis of data on the genes of recessive diseases. A matrix of Nei's genetic distances was calculated from the frequencies of 45 recessive diseases found in the seven districts studied. The average genetic distance calculated for the 45 loci of autosomal recessive diseases was 0.006175 x 10(-3). Similarly, matrix of genetic distances for five Mari populations was obtained (Medvedevskii and Zvenigovskii raions were not included) based on a total of 32 allelic frequencies for ten polymorphic immune and biochemical loci. The average genetic distance calculated from the ten polymorphic loci was 0.001930, i.e., 2.5 orders of magnitude greater than the average genetic distance for recessive diseases. The matrices of genetic distances for the five Mari populations calculated from the gene frequencies for recessive diseases and for the ten polymorphic systems were largely similar to each other. Thus, the main elements of the genetic structure of the Mari population can be estimated on the basis of gene frequencies for hereditary diseases. In this case, the characteristics of individual populations, which are more or less isolated, and of their interaction are the same as in the case of studying genetic structure with the use of polymorphic biological markers. PMID- 9749340 TI - [Genetic characteristics of the population of Severo-Baikal'skii region of the northern part of the Buryat Republic]. AB - In the populations of seven settlements of Severo-Baikal'skii raion (Buryat Republic), 12 genetic systems were studied: ABO, Rh, MN, P, Lewis, HP, GC, TF, PI, Gm1, ABH, and the cerumen consistency. With respect to most of these genetic systems, the studied populations were either insignificantly different from other Siberian populations or intermediate between Russian populations from the European part of Russia and from indigenous Cis-Baikal populations (Buryats and Evenks). This might be related to outbreeding and selection. The observed parameters of genetic differentiation indicated that the studied system of subpopulations was at an equilibrium, with insignificant differences between subpopulations. PMID- 9749341 TI - [Variability of pericentromeric chromatin in chromosome 2 of ovarian nurse cells during inbreeding in Anopheles atroparvus V.Tiel]. AB - We investigated the variability of pericentromeric chromatin of chromosome 2 in ovarian nurse cells (trophocytes) in two laboratory lines of malaria mosquito Anopheles atroparvus V. Tiel and in their hybrids. One line had been raised by means of sib inbreeding, the other kept at constantly high population density. The inbreeding was shown to result in an increased percentage of chromosomes bearing an achromatinic zone in the centromeric region, which resulted in chromosome breakage. Toxicological tests demonstrated an increase in the sensitivity of the progeny of females with abnormal morphotypes of chromosome 2 to the entomopathogenic bacterium Bacillus thuringiensis israelensis. The appearance of the achromatinic zone is attributed to local chromatin underreplication accompanying chromosome polytenization. Possible reasons for this phenomenon and its implication for adaptation are discussed. PMID- 9749342 TI - [Differentiation of domestic horse and Przewalskis horse using various DNA sequences]. AB - The electrophoretic mobility of seven erythrocyte enzymes and spectra of fragments amplified by RAPD-PCR with primers UBC-85 and UBC-126 were comparatively analyzed in domestic horse and Przewalski's horse. All tested genetic markers were classified into two groups differing in their involvement in differentiation of the two closely related horse species. Markers from different groups differed neither in their type (a polymorphic protein or an amplification product) nor in their biochemical role (for enzymes). PMID- 9749343 TI - [Interpopulation diversity of the gene pool: beta distribution of Wright's F(ST) statistics]. AB - The distribution of FST(i) estimates were studied in representative samples of ith genes for the gene pools of the major human populations, including the populations of Europe, Asia, Africa, Australia, America, North-eastern Eurasia, and five subregions of the latter. An average of 80 FST(i) estimates were analyzed for each sample of marker genes with a level of polymorphism (q) from 0.05 to 0.95. For each gene pool, the empirical distributions of FST(i) estimates were approximated by the main types of theoretical distributions--the normal, chi 2, Weibull, gamma, and beta distributions. In all gene pools, only beta distributions were good approximations of the empirical FST(i) distributions. The parameters of the beta distributions are reported in this study. It was demonstrated that the characteristics of beta distributions for all major gene pools studied could be interpolated to the smaller constituent gene pools. since the use of the traditional parametric tests starts from an assumption of normal distribution, they are inapplicable to analysis of FST statistics. Therefore, the obtained parameters of beta distributions should be used. These parameters allow the confidence intervals of the average FST values to be determined and permit correct comparison between the characteristics of both individual genes and gene pools to be performed. PMID- 9749346 TI - [Eukaryotic DNA topoisomerase and in vitro recombination between circular supercoiled plasmid DNA's in an in vitro system]. AB - Eukaryotic DNA topoisomerase II was shown to catalyze recombination between circular supercoiled plasmid DNAs in vitro. The results obtained are discussed with regard to the organization of eukaryotic chromosomes in the form of topologically independent domains (loops). PMID- 9749344 TI - [Mitochondria DNA markers and genetic demographic processes in neolithic Europe]. AB - Distribution patterns of mitochondrial DNA markers, BamHI-2/AvaII-5(3), BamHI 3/MspI-4, and BamHI-1/AvaII-5(3), and mitotypes with the control region corresponding to the Cambridge reference sequence in European and Middle Eastern populations are discussed with respect to the history of the European populations in the Neolith. It is suggested that distribution of mtDNA markers is associated with the Neolithic invasion of the Caucasoid populations from the Middle East to Europe in accordance with the "wave-of-advance" hypothesis of Ammerman and Cavalli-Sforza. However, genetic differences between regional European population groups indicate a more pronounced demic diffusion in the southeast of Europe, better correlated with the hypothesis of C. Renfrew, while in the northeast the agriculture and the Indo-European languages could have been adopted by the ancient Caucasoid populations without any considerable genetic admixture with at least the women from the Neolithic migration wave from the Middle East. PMID- 9749345 TI - [Action of low doses of gamma-radiation on cytogenetic damage in polychromatophilic erythrocytes of bone marrow in mice in vivo]. AB - The frequency of micronuclei was estimated in bone marrow polychromatic erythrocytes (PCEs) of mice gamma-irradiated in vivo at doses of 5-50 cGy. The dose-effect curve was found to have a complex stepwise pattern with three consequent segments: (1) high radiosensitivity, (2) a significant decrease in radiosensitivity, and (3) an increase in radiosensitivity. PMID- 9749348 TI - Involvement of angiotensin II receptor subtypes during testicular development in rats. AB - Expression of testicular angiotensin II (AT2) receptors in Sprague-Dawley rats at various stages of development (1 and 5 days, 2, 3, 4 and 7 weeks postnatal) were studied by in vitro autoradiography and Northern blot analysis. The receptors were labelled with 125I-[Sar1, Ile8]AT2 and differentiated into two subtypes according to their susceptibility to AT1 (losartan, 5 microM) or AT2 (PD123319, 5 microM) antagonist. Total AT2 receptor binding in the testis was highest at 1 day of age (8.12 +/- 0.35 fmol/mg protein, mean +/- secEM, n = 8) and decreased gradually thereafter (5 days: 6.9 +/- 0.41, 2 weeks: 2.85 +/- 0.10, 3 weeks: 1.64 +/- 0.19, 4 weeks: 0.76 +/- 0.09, 6 weeks: 0.77 +/- 0.09 fmol/mg protein, n = 8 11). AT2 receptor binding was strikingly abundant in 1-day-old rat testis (6.98 +/- 0.34 fmol/mg protein), while considerably less AT1 receptor binding (1.46 +/- 0.19 fmol/mg protein) was observed. The relative amounts of each subtype did not change for the first 3 weeks but the 4-week-old rat testis contained almost exclusively AT1 receptors (0.63 +/- 0.05 fmol/mg protein). Northern blot analysis showed that mRNA expression of both AT1 and AT2 types decreased with age. Microscopic emulsion autoradiography was undertaken to clarify the localization of binding. At 10 days of age, both AT1 and AT2 receptors were present in the interstitial area, whereas seminiferous tubules contained mainly AT2 receptors. At 7 weeks of age, no significant binding was observed in the seminiferous tubule and the interstitial area contained AT1 receptors exclusively. These results demonstrate expression of AT2 receptors in the rapidly growing testis and suggest that change in the levels of AT2 receptor subtypes may be relevant to development and/or growth of the testis. PMID- 9749349 TI - Transurethral resection of cystic and non-cystic ejaculatory duct obstructions. AB - Ejaculatory duct obstructions are diagnosed in approximately 5% of azoospermic men and can be treated by transurethral resection (TURED) or incision of the ducts. Eight patients with azoospermia and ejaculatory duct obstructions were treated by TURED after clinical examination, semen analysis, biochemical analysis of seminal plasma, endocrine analysis, transrectal ultrasonography and testicular biopsy. In 3/3 cases of cystic and in 3/5 cases of non-cystic obstruction. TURED of the stenosis was possible. During a follow-up of 12 months there was an increase in semen volume and sperm count in 3/3 and 3/5 patients, respectively. No pregnancy was achieved during the period up to 12 months. Clinical symptoms such as haemospermia and pain disappeared in all cases. In our cases and another 98 cases of ejaculatory duct obstructions documented in the literature, men of semen quality improved in 38-60% with a pregnancy rate of men 22-31% after TURED. We conclude that there is a correlation between the aetiology of ejaculatory duct obstructions and success rate of TURED. PMID- 9749347 TI - Pituitary-dependent expression of the testicular angiotensin II receptor and its subtypes in rats. AB - Angiotensin II (AT2) has been implicated in the growth and/or differentiation of its target tissues. In the present study, testicular AT2 receptor and its subtypes in hypophysectomized rats were examined using quantitative in vitro autoradiography and Northern blot analysis in an attempt to determine possible involvement of pituitary hormones in their expression. Prepubescent (3 weeks of age) male Sprague-Dawley rats underwent hypophysectomy or sham operation. From 10 days thereafter, they were treated with vehicle, growth hormone, human chorionic gonadotrophin or human menopausal gonadotrophin for 10 days. Testicular AT2 receptors were labelled with 125I-[Sar1,Ile8] AT2 and differentiated into its subtypes (AT1 and FAT2) according to their susceptibility to AT1 (losartan, 5 microM) and AT2 (CGP42112B, 1 microM) antagonists. Hypophysectomy led to a marked increase in AT2 receptor concentration (sham-operated rats: 0.7 +/- 0.2 fmol/mg protein, hypophysectomized rats: 2.5 +/- 0.6 fmol/mg protein, mean +/- SEM, n = 11-12, p < 0.01) with predominant occurrence of AT1 receptors. Both human chorionic gonadotrophin and human menopausal gonadotrophin decreased testicular AT2 receptor concentration, whereas growth hormone did not affect AT2 receptor expression. Northern blot analysis revealed both testicular AT1 and AT2 receptor mRNA expression to be significantly increased after hypophysectomy and reduced by gonadotrophin treatment. These results suggest that the expression of testicular AT2 receptors is regulated by pituitary gonadotrophins and that AT2 may play a role in testicular growth and/or differentiation. PMID- 9749350 TI - Mouse spermatozoa modify their motility parameters and chemotactic response to factors from the oocyte microenvironment. AB - The aim of this work was to evaluate the period of time required for the induction of changes in motility of mouse spermatozoa in response to factors from the microenvironment of oocytes. To determine the effects of the latency time, the period of preincubation before contact with the oocyte product(s), sperm samples were incubated for 15 or 90 min and then exposed to either Dulbecco's Modified Eagle's Medium (DMEM) or to a crude extract of superovulated oocytes (CE). The assays were performed in a Zigmond chamber by filling one compartment with either DMEM or CE, and the other compartment with the sperm suspension. A videomicroscopy system was used for tracking the spermatozoa. Sperm motility analysis was assessed using a semiautomatic objective method, and the following parameters were determined; (1) dynamic parameters: curvilinear velocity, linear velocity and linearity; (2) progressive motility: percentage of spermatozoa showing either circular or linear patterns of movement; and (3) directional motility: percentage of spermatozoa that moved towards either the DMEM or the CE. The results of this work showed that when the spermatozoa contacted soluble factors in CE after only 15 min of previous incubation, there was a significant increase in their dynamic parameters, change in their progressive motility, and induction of directional movement of the spermatozoa towards the CE components, while a longer period of preincubation did not significantly modify these effects. On the other hand, in the presence of culture medium (with or without addition of bovine serum albumin), the spermatozoa needed a more extended period of incubation to significantly increase their dynamic parameters and to modify their progressive motility, while maintaining a random direction of movement. PMID- 9749351 TI - Sperm characteristics and zona pellucida binding in relation to field fertility of frozen-thawed semen from dairy AI bulls. AB - The present study examined the relationship between bull sperm characteristics immediately post-thaw and some characteristics registered after swim-up, including the ability of spermatozoa to bind to homologous zona pellucidae (ZP) in vitro, and fertility after artificial insemination (AI) of 9426 females. Frozen-thawed semen from 22 AI bulls of the Swedish Red and White Breed, represented by 43 different frozen batches (1-4 batches/bull, 2 consecutive ejaculates/batch), was examined with the aim of determining concentration, motility patterns, morphology and membrane integrity. In addition, the frozen thawed spermatozoa were subjected to a swim-up procedure and those separated in this way were tested with two assays of sperm-binding to the ZP of homologous oocytes in vitro (ZBA), using either a relative ZBA index against a control bull of proven high fertility or absolute binding (Absolute ZBA). The correlations of the various sperm traits and 56-day non-return rates (NRR) after field AI were retrospectively examined as single traits and as combinations of traits (combined measures), including regression analysis of significant traits. Among the sperm characteristics, positively significant (p < 0.01) correlations with NRR were found for linear motility post-thawing (r = 0.45-0.59) and the concentration of motile spermatozoa after swim-up (r = 0.43-0.63). Results obtained with the absolute ZBA approach were significantly (p < 0.05) correlated with NRR (r = 0.50), whereas the correlation between NRR and the ZBA index was not significant. The use of combined measures of sperm traits, including the ability to bind to ZP, showed a stronger predictive correlation with NRR (r = 0.68-0.75), compared with single traits. The results suggest that the combined analysis of sperm linear-motility patterns, swim-up separated sperm motility and absolute ZBA can provide a valuable assessment of the fertilizing capacity of AI bull semen. PMID- 9749354 TI - Zinc enrichment in prostasomes. PMID- 9749352 TI - The appearance of basal cells in the developing murine epididymis and their temporal expression of macrophage antigens. AB - This work demonstrates similarities between epididymal basal cells and macrophages in the mouse. Light microscopic studies of the postnatal development of the murine epididymis showed that basal cells were not present before days 12, 14 and 16 in the cauda, caput and corpus epididymis, respectively. An increase in cell number per unit length of tubule perimeter was demonstrated in all segments between days 20 and 27, when testicular fluid and spermatozoa start entering the epididymis. In the adult, there were more basal cells per unit perimeter in the cauda than caput or corpus epididymis. Conspicuous and consistent expression by basal cells of antigens detected by antibodies against tissue-fixed macrophages (F4/80) and mature macrophages (Mac-1) occurred only after they became established within the epithelium. Basal cells in the cauda epididymis did not display either antigen in the adult, although they persisted in the caput region. Such developmental patterns are compatible with the hypothesis that basal cells play a role in immune defence against sperm autoantigens. PMID- 9749353 TI - Prognostic value of male diagnostic profiles in intracytoplasmic sperm injection (ICSI). AB - Possible correlations between male hormone and semen parameters with pregnancy and oocyte fertilization rates following intracytoplasmic sperm injection (ICSI) were investigated. The study is based on 290 couples who underwent ICSI therapy for the first time. The parameters evaluated were male age, serum levels of follicle stimulating hormone (FSH) and testosterone, sperm concentration, sperm motility, normal sperm morphology, index of teratozoospermia (TZI) and sperm vitality. A marginal, barely significant association was found between the fertilization rate and serum FSH levels in the male partner (p = 0.046). There was no relevant association between male parameters and pregnancy rates. The study confirms that male hormonal and semen parameters are of low prognostic value for the outcome of ICSI. PMID- 9749355 TI - Pretelangiectasis and telangiectasis of the bovine liver: a morphological, immunohistochemical and ultrastructural study. AB - Forty-five livers from conventionally slaughtered Holstein-Friesian steers with telangiectasis were studied by histochemical methods, immunolabelling for fibronectin, laminin and type IV collagen, and transmission electron microscopy. None of the previously described changes in telangiectasis (necrosis, hepatitis, thromboembolism, dilatation of the space of Disse by glycogen extruded from hepatocytes and reduced density of the perisinusoidal reticulin framework) were evident. Pretelangiectasis (sinusoidal dilatation) and telangiectasis (blood filled cavities) were characterized by sinusoidal barrier alterations, leading to sinusoidal capillarization; and there was progressive formation of a true basement membrane and perisinusoidal fibrosis. Comparison of bovine liver telangiectasis and human peliosis hepatis suggests that they have a similar pathogenesis. It is suggested that a primary alteration of the sinusoidal barrier is responsible for an increased deposition of basement membrane components (fibronectin, laminin, type IV collagen) in the perisinusoidal region, and fibrosis. These are likely to render the exchange of oxygen and substrates between blood and hepatocytes more difficult and to produce haemodynamic imbalances, leading to hepatocyte atrophy and eventually to sinusoidal disruption. PMID- 9749356 TI - Immunohistochemical and ultrastructural evidence of hog cholera virus infection of megakaryocytes in bone marrow and spleen. AB - Twelve pigs were inoculated with a highly virulent strain of hog cholera virus (HCV) to study viral infection of megakaryocytes in the bone marrow and spleen. Immunohistochemical and ultrastructural examination revealed HCV infection in a small proportion (2.5-9.0%) of these cells from the 2nd to the 9th day after inoculation, at which time the experiment was terminated. Megakaryocyte infection accounts for the presence of viral antigens in platelets. The latter may represent a passive vehicle for spreading the virus in the animal. PMID- 9749357 TI - Lymphadenopathy and non-suppurative meningo-encephalitis in calves experimentally infected with bovine immunodeficiency-like virus (FL112). AB - In an experiment on bovine immunodeficiency-like virus (BIV), the virological and serological aspects of which were reported in an earlier paper, three groups (A, B and C) of three calves were inoculated subcutaneously with a recently isolated strain (FL112). For group B and group C, the virus was suspended in milk, and for group C (controls) the viral suspension was subjected to pasteurization before inoculation. The calves were killed for necropsy 12 months later. Clinical assessment revealed subtle ataxia in two group A calves, which took the form of an intermittent "shifting" (from one leg to another) lameness, and palpable enlargement of the pre-scapular lymph nodes in one group B animal. At necropsy, haemal lymph nodes (0.1 to 0.5 cm in diameter), occurring singly, were observed in all animals. However, in groups A and B (but not C), enlarged haemal lymph nodes (< or = 2 cm in diameter) were also seen, occurring singly and in chains; and in one group A animal they occurred in grape-like clusters. In groups A and B (but not C), histopathological examination revealed generalized hyperplastic changes in lymph nodes, especially the haemal lymph nodes. This finding was particularly striking in the two clinically ataxic animals from group A, which also showed a non-suppurative meningo-encephalitis; the latter was possibly the cause of the subtle clinical signs. This study supports previous findings on lymphadenopathy resulting from experimental infection with BIV. PMID- 9749358 TI - Immunohistochemical characterization of the lesions of feline progressive lymphocytic cholangitis/cholangiohepatitis. AB - The histopathological features of liver biopsies from 20 cats with progressive lymphocytic cholangitis/cholangiohepatitis are reported. These biopsies were subject to immunohistochemical investigation for expression of CD3, CD79. Major Histocompatibility Complex (MHC) Class II molecules, and feline IgG, IgM and IgA. Livers from five normal cats, which were also examined showed constitutive expression of MHC Class II by sinusoidal Kupffer cells and bile duct epithelium, in addition to a population of portal, and bile duct inter-epithelial, CD3+ T lymphocytes. In liver biopsies from cats with the active phase of lymphocytic cholangitis/cholangiohepatitis (n = 11), the portal lymphocytes were predominantly CD3+ T cells that infiltrated bile duct epithelium and periportal hepatic parenchyma, CD79+ B lymphocytes formed distinct aggregates or follicles within the regions of T-cell infiltration. Low numbers of plasma cells were present, and these predominantly expressed IgA. MHC Class II was expressed by Kupffer cells, infiltrating T and B lymphocytes and macrophages. There was membrane and cytoplasmic Class II expression by bile duct epithelium, some vascular endothelium, and fibroblasts within areas of fibrosis. In liver biopsies from cats with chronic lymphocytic cholangitis/cholangiohepatitis (n = 9), there was less in flammation, but the composition of the infiltrates was similar to that in the active phase of disease. The findings provide further evidence for an immune mediated pathogenesis in progressive lymphocytic cholangitis/cholangiohepatitis. PMID- 9749359 TI - Morphological effects of Pasteurella multocida type-D dermonecrotoxin on rat osteosarcoma cells in a nude mouse model. AB - Of 15 athymic nude mice that received subcutaneous implants of a rat osteosarcoma cell line, two groups of four subsequently received either a short (group 1) or a more prolonged (group 2) course of subcutaneous injections of the dermonecrotic toxin (DNT) of Pasteurella multocida type D. The remaining seven mice (controls) received no DNT. Both groups of DNT-treated mice lost body weight as compared with controls. Tumour weight, expressed as a percentage of body weight, increased in the four group 1 mice. Tumours in this group 1 were consistently larger than those in appropriate controls, indicating that this percentage was not simply a function of decreased body weight. The immunohistochemical labelling of proliferating cell nuclear antigen (PCNA) and morphometric analysis of intratumoral necrosis suggested that the DNT had a mitogenic effect and contributed to the neoplastic growth. The presence of foci of neoplastic osteoblasts in the lungs of some DNT-treated mice suggested that the enhanced tumour growth led to an increased incidence of metastasis. PMID- 9749360 TI - Neuropathological study of gazelle herpesvirus 1 (equine herpesvirus 9) infection in Thomson's gazelles (Gazella thomsoni). AB - Gazelle herpesvirus (GHV-1), correctly designated as equine herpesvirus 9, is a new type of equine herpesvirus immunologically related to equine herpesvirus 1 (EHV-1). As a sequel to a virological study, the neuropathology of encephalitis caused by GHV-1 in Thomson's gazelles (Gazella thomsoni) was examined. Seven gazelles died with or without neurological symptoms between early September and mid-October in 1993. No gross abnormalities were observed at necropsy, but all animals had non-suppurative encephalitis, characterized by necrosis and degeneration of neurons, glial reactions and perivascular cuffing in the cerebrum. Five cases showed intranuclear inclusion bodies, with the appearance of herpesvirus in the degenerating neurons. Immunohistochemically, all seven animals showed a positive reaction to EHV-1 antigen in neurons in the necrotic areas of the cortex. The clinical course and morphological features of GHV-1 encephalitis were distinct from those of EHV-1-induced encephalitis in the horse, which is characterized by vasculitis, thrombosis, ischaemia, and lack of intranuclear inclusions in neurons. PMID- 9749361 TI - Detection and distribution of DNA of Actinobacillus pleuropneumoniae in the lungs of naturally infected pigs by in-situ hybridization. AB - Detection and distribution of Actinobacillus pleuropneumoniae was studied in formalin-fixed paraffin wax-embedded lung tissues from 10 naturally infected pigs by in-situ hybridization with a non-radioactive digoxigenin-labelled probe. A 610 base pair cDNA probe from a genomic library of A. pleuropneumoniae was generated by the polymerase chain reaction. All 10 pigs infected with A. pleuropneumoniae serotypes 2, 5, 6, or an untypable strain showed a distinct, positive signal in the degenerate alveolar leucocytes bordering a zone of coagulative necrosis and in the dense zone of degenerated cells in granulation tissue surrounding the necrotic areas. Positive cells typically exhibited dark-brown to black labelled deposits without background staining. A. pleuropneumoniae nucleic acids were more readily detected in areas of coagulative necrosis than in areas of granulation tissue. In-situ hybridization demonstrated that A. pleuropneumoniae primarily infected neutrophils and alveolar macrophages. The technique used was rapid, specific and sensitive, and may prove useful for the diagnosis of A. pleuropneumoniae infection in routinely fixed and processed tissues, obviating the need for bacterial isolation. PMID- 9749362 TI - Optimization of the immunohistochemical demonstration of keratins in paraffin wax embedded mouse skin. AB - The purpose of this study was to develop an immunoperoxidase technique for the detection of cytokeratins in samples of paraffin wax-embedded adult and fetal skin from NMRI mice, with various antibodies (Troma-1, LL001, 8.60, MCK5, MCK6, AF129) that have been tested mainly on fresh-frozen sections. Each antibody was tested with three different fixatives (10% neutral buffered formalin, Bouin's fluid, and 70% ethanol) and two distinct pretreatments (enzymatic digestion with trypsin, or heat treatment). The best results, in terms of non-specific background labelling, morphological preservation and intensity of specific labelling, were obtained (1) for adult skin, by the use of Bouin's fluid, heat pretreatment and antibodies LL001, MCK5, MCK6 or AF129, and (2) for fetal skin, by the use of 70% ethanol, heat pretreatment and antibody Troma-1. Monoclonal antibody 8.60 gave the best results when the use of 70% ethanol was combined with either enzymatic digestion or heat pretreatment. PMID- 9749363 TI - Adrenal cortical epithelial cysts in two saddleback tamarins (Saguinus fuscicollis). AB - Adrenal cysts of epithelial origin were found incidentally in the adrenal cortical tissues of two adult female saddleback tamarins. In one case, a small cluster of minor cysts was located at the cortico-medullary border. The cysts were filled with a periodic acid-Schiff (PAS)-positive, amorphous substance and lined by a cuboidal PAS-negative epithelium, which resembled morphologically the ascending Henle's loops of the kidney. These microcysts were thought to be mesonephric remnants. In the second case, two large cysts of 1 to 3 mm diameter extended throughout the entire cortex. The cysts were filled by a watery, PAS negative fluid and lined by a bi- to multi-layered, cytokeratin-positive epithelium. The basal epithelial layer consisted of cuboidal cells, which became cylindrical to drop-like in appearance towards the cyst lumen. The cysts closely resembled mesothelium-derived adrenal cysts in man. This is the first report of adrenal cysts in non-human primates. PMID- 9749364 TI - Malignant melanoma of the nictitating membrane in a cat (Felis vulgaris). AB - A case of malignant melanoma of the nictitating membrane in a 10-year-old cat is described. The primary unilateral tumour mass appeared to originate from the surface of the nictitating membrane, with infiltrative growth in the orbital cavity, resulting in exophthalmia. No ocular involvement was noted. Metastatic spread was seen in the cerebrum and the lungs. To our knowledge, this is the first reported case of a metastasizing melanoma originating from the nictitating membrane in a cat. PMID- 9749365 TI - Granular cell foci of the uterus in Donryu rats. AB - Eleven cases of uterine granular cell foci were observed in a total of 855 female Donryu rats. All the lesions were microscopical and focal or multifocal in nature, and composed of uniformly large cells with abundant granulated eosinophilic cytoplasm. Histopathologically, immunohistochemically and ultrastructurally they resembled granular cell tumours of the uterus reported in rats and mice. The development of granular cell foci may be the result of hormonal stimulation. PMID- 9749366 TI - The application of three approximate free energy calculations methods to structure based ligand design: trypsin and its complex with inhibitors. AB - Three approximate free energy calculation methods are examined and applied to an example ligand design problem. The first of the methods uses a single simulation to estimate the relative binding free energies for related ligands that are not simulated. The second method is similar, except that it uses only first derivatives of free energy with respect to atomic parameters (most often charge, van der Waals equilibrium distance, and van der Waals well depth) to calculate free energy differences. The last method PROFEC (Pictorial Representation of Free Energy Components), generates contour maps that show how binding free energy changes when additional particles are added near the ligand. These three methods are applied to a benzamidine/trypsin complex. They each reproduce the general trends in the binding free energies, indicating that they might be useful for suggesting how ligands could be modified to improve binding and, consequently, useful in structure-based drug design. PMID- 9749367 TI - Structure-based design of ligands for protein basic domains: application to the HIV-1 Tat protein. AB - A methodology has been developed for designing ligands to bind a flexible basic protein domain where the structure of the domain is essentially known. It is based on an empirical binding free energy function developed for highly charged complexes and on Monte Carlo simulations in internal coordinates with both the ligand and the receptor being flexible. HIV-1 encodes a transactivating regulatory protein called Tat. Binding of the basic domain of Tat to TAR RNA is required for efficient transcription of the viral genome. The structure of a biologically active peptide containing the Tat basic RNA-binding domain is available from NMR studies. The goal of the current project is to design a ligand which will bind to that basic domain and potentially inhibit the TAR-Tat interaction. The basic domain contains six arginine and two lysine residues. Our strategy was to design a ligand for arginine first and then a superligand for the basic domain by joining arginine ligands with a linker. Several possible arginine ligands were obtained by searching the Available Chemicals Directory with DOCK 3.5 software. Phytic acid, which can potentially bind multiple arginines, was chosen as a building block for the superligand. Calormetric binding studies of several compounds to methylguanidine and Arg-/Lys-containing peptides were performed. The data were used to develop an empirical binding free energy function for prediction of affinity of the ligands for the Tat basic domain. Modeling of the conformations of the complexes with both the superligand and the basic domain being flexible has been carried out via Biased Probability Monte Carlo (BPMC) simulations in internal coordinates (ICM 2.6 suite of programs). The simulations used parameters to ensure correct folding, i.e., consistent with the experimental NMR structure of a 25-residue Tat peptide, from a random starting conformation. Superligands for the basic domain were designed by joining together two molecules of phytic acid with peptidic and peptidomimetic linkers. The linkers were refined by varying the length and side chains of the linking residues, carrying out BPMC simulations, and evaluation of the binding free energy for the best energy conformation. The dissociation constant of the best ligand designed is estimated to be in the low- to mid-nanomolar range. PMID- 9749368 TI - Dihydrofolate reductase: a potential drug target in trypanosomes and leishmania. AB - Dihydrofolate reductase has successfully been used as a drug target in the area of anti-cancer, anti-bacterial and anti-malarial chemotherapy. Little has been done to evaluate it as a drug target for treatment of the trypanosomiases and leishmaniasis. A crystal structure of Leishmania major dihydrofolate reductase has been published. In this paper, we describe the modelling of Trypanosoma cruzi and Trypanosoma brucei dihydrofolate reductases based on this crystal structure. These structures and models have been used in the comparison of protozoan, bacterial and human enzymes in order to highlight the different features that can be used in the design of selective anti-protozoan agents. Comparison has been made between residues present in the active site, the accessibility of these residues, charge distribution in the active site, and the shape and size of the active sites. Whilst there is a high degree of similarity between protozoan, human and bacterial dihydrofolate reductase active sites, there are differences that provide potential for selective drug design. In particular, we have identified a set of residues which may be important for selective drug design and identified a larger binding pocket in the protozoan than the human and bacterial enzymes. PMID- 9749369 TI - PAPQMD parametrization of molecular systems with cyclopropyl rings: conformational study of homopeptides constituted by 1-aminocyclopropane-1 carboxylic acid. AB - The suitability of ab initio, semiempirical and density functional methods as sources of stretching and bending parameters has been explored using the PAPQMD (Program for Approximate Parametrization from Quantum Mechanical Data) strategy. Results show that semiempirical methods provide parameters comparable to those compiled on empirical force fields. In this respect the AMI method seems to be a good method to obtain parameters at a minimum computational cost. On the other hand, harmonic force fields initially developed for proteins and DNA have been extended to include compounds containing highly strained three-membered rings, like 1-aminocyclopropane-1-carboxylic acid. For this purpose the cyclopropyl ring has been explicitly parametrized at the AMI level considering different chemical environments. Finally, the new set of parameters has been used to investigate the conformational preferences of homopeptides constituted by 1-aminocyclopropane-1 carboxylic acid. Results indicate that such compounds tend to adopt a helical conformation stabilized by intramolecular hydrogen bonds between residues i and i + 3. This conformation allows the arrangement of the cyclic side chains without steric clashes. PMID- 9749370 TI - Molecular modeling of the human vasopressin V2 receptor/agonist complex. AB - The V2 vasopressin renal receptor (V2R), which controls antidiuresis in mammals, is a member of the large family of heptahelical transmembrane (7TM) G protein coupled receptors (GPCRs). Using the automated GPCR modeling facility available via Internet (http:/(/)expasy.hcuge.ch/swissmod/SWISS-MODEL.+ ++html) for construction of the 7TM domain in accord with the bovine rhodopsin (RD) footprint, and the SYBYL software for addition of the intra- and extracellular domains, the human V2R was modeled. The structure was further refined and its conformational variability tested by the use of a version of the Constrained Simulated Annealing (CSA) protocol developed in this laboratory. An inspection of the resulting structure reveals that the V2R (likewise any GPCR modeled this way) is much thicker and accordingly forms a more spacious TM cavity than most of the hitherto modeled GPCR constructs do, typically based on the structure of bacteriorhodopsin (BRD). Moreover, in this model the 7TM helices are arranged differently than they are in any BRD-based model. Thus, the topology and geometry of the TM cavity, potentially capable of receiving ligands, is in this model quite different than it is in the earlier models. In the subsequent step, two ligands, the native [arginine8]vasopressin (AVP) and the selective agonist [D arginine8]vasopressin (DAVP) were inserted, each in two topologically non equivalent ways, into the TM cavity and the resulting structures were equilibrated and their conformational variabilities tested using CSA as above. The best docking was selected and justified upon consideration of ligand-receptor interactions and structure-activity data. Finally, the amino acid residues were indicated, mainly in TM helices 3-7, as potentially important in both AVP and DAVP docking. Among those Cys112, Val115-Lys116, Gln119, Met123 in helix 3; Glu174 in helix 4; Val206, Ala210, Val213-Phe214 in helix 5; Trp284, Phe287 Phe288, Gln291 in helix 6; and Phe307, Leu310, Ala314 and Asn317 in helix 7 appeared to be the most important ones. Many of these residues are invariant for either the GPCR superfamily or the neurophyseal (vasopressin V2R, V1aR and V1bR and oxytocin OR) subfamily of receptors. Moreover, some of the equivalent residues in V1aR have already been found critical for the ligand affinity. PMID- 9749371 TI - A flexible triangulation method to describe the solvent-accessible surface of biopolymers. AB - A relatively simple protein solvent-accessible surface triangulation method for continuum electrostatics applications employing the boundary element method is presented. First, the protein is placed onto a three-dimensional lattice with a specified lattice spacing. To each lattice point, a box is assigned. Boxes located in the solvent region and in the interior of the protein are removed from the set. Improper connections between boxes and the possible existence of cavities in the interior of the protein which would destroy the proper connectivity of the triangulated surface are taken care of. The remaining set of boxes define the outer contour of the protein. Each free face exposed to the solvent of the remaining set of boxes is triangulated after the surface defined by the free faces has been smoothed to follow the shape of the macromolecule more accurately. The final step consists of a mapping of triangle vertices onto a set of surface points which define the solvent-accessible surface. Normal vectors at triangle vertices are obtained also from the free faces which define the orientation of the surface. The algorithm was tested for six molecules including four proteins; a dipeptide, a helical peptide consisting of 25 residues, calbindin, lysozyme, calmodulin and cutinase. For each molecule, total areas have been calculated and compared with the result computed from a dotted solvent accessible surface. Since the boundary element method requires a low number of vertices and triangles to reduce the number of unknowns for reasons of efficiency, the number of triangles should not be too high. Nevertheless, credible results are obtained for the total area with relative errors not exceeding 12% for a large lattice spacing (0.30 nm) while close to zero for a smaller lattice spacing (down to 0.16 nm). The output of the triangulation computer program (written in C++) is rather simple so that it can be easily converted to any format acceptable for any molecular graphics programs. PMID- 9749372 TI - A semiempirical study on inhibition of catechol O-methyltransferase by substituted catechols. AB - Catechol and endogenous catechol derivatives are readily methylated by catechol O methyltransferase (COMT). In contrast, many catechol derivatives possessing electronegative substituents are potent COMT inhibitors. The X-ray structure of the active site of COMT suggests that the methylation involves a lysine as a general base. The lysine can activate one of the catecholic hydroxyl groups for a nucleophilic attack on the active methyl group of the coenzyme S-adenosyl-L methionine (AdoMet). We studied the effect of dinitrosubstitution of the catecholic ring at the semiempirical PM3 level on the methylation reaction catalysed by COMT. The electronegative nitro groups make the ionized catechol hydroxyls less nucleophilic than the corresponding hydroxyl groups of the non substituted catechol. As a consequence, dinitrocatechol is not methylated but is instead a potent COMT inhibitor. The implications of this mechanism to the design of COMT inhibitors are discussed. PMID- 9749373 TI - Bile salt deconjugation and cholesterol removal from media by Lactobacillus casei. AB - Lactobacillus casei N19 and E5 and Lactobacillus acidophilus L1 and ATCC 43121 were compared for their ability to deconjugate bile salts and remove cholesterol from MRS broth during growth at pH 6.0 and during growth without pH control. Samples grown without pH control dropped to pH 4.2 to 4.5 during 20 h of incubation, depending on the culture. The plate counts indicated that populations in all cultures were near their maximum numbers after 16 h of growth. The amount of cholesterol removed from the broth was similar for both strains of L. acidophilus grown with and without pH control. However, the strains of L. casei differed significantly in the amount of cholesterol removed during growth with or without pH control. Both cultures of L. casei that were grown at pH 6.0 removed very little cholesterol from the broth, but cells grown without pH control removed up to 60 micrograms of cholesterol/ml. All cultures of both species deconjugated 60 to 90% of the bile salts. Lactobacillus acidophilus L1 was the only culture to demonstrate differences between the two pH treatments in the amount of bile salts deconjugated; however, there was no difference in the amounts of cholesterol removed. These results indicate that most of the cholesterol removal from broth by L. acidophilus was due to assimilation, perhaps by the incorporation of cholesterol into the cellular membrane. Lactobacillus casei most likely removes cholesterol from broth by means of the destabilization of cholesterol micelles and the coprecipitation of the cholesterol with the deconjugated bile salts at pH less than 6.0. PMID- 9749375 TI - Changes in physicochemical properties of retort-sterilized dairy beverages during storage. AB - The effects of composition, storage time, and storage temperature on the physicochemical properties of a retort-sterilized dairy beverage were investigated. Drinks with eight formulations were stored at 4, 25, and 37 degrees C for 6 mo and were analyzed monthly for pH, net color difference, apparent viscosity, sedimentation index, homogenization index, particle size index, and soluble calcium. The changes in the physicochemical properties of the beverages increased as storage time and temperature increased. The degree of change was affected by the composition of the product. Sodium tripolyphosphate was implicated in promoting age gelation of samples with 11% nonfat milk solids, but sedimentation was observed in the absence of sodium tripolyphosphate and carrageenan. The apparent viscosity of samples affected the rate of age gelation and sedimentation, both of which increased as viscosity decreased. Interactions between milk fat, carrageenan, and nonfat milk solids were important in determining the apparent viscosity of the beverages and the rate of change observed during storage. PMID- 9749374 TI - The primary structure of caprine PP3: amino acid sequence, phosphorylation, and glycosylation of component PP3 from the proteose-peptone fraction of caprine milk. AB - Proteose-peptone component 3 is a phosphorylated glycoprotein that was isolated from the proteose-peptone fraction of caprine milk. By mass spectrometric analysis, amino acid sequencing, and polymerase chain reaction analysis, the primary structure has been determined and has been shown to contain 136 amino acids. Phosphorylations were identified at Ser30 and Ser41. A partial glycosylation was present at Thr16, and a N-linked glycosylation was present at Asn78. Galactosamine was the amino sugar detected at Thr16. Glucosamine and galactosamine were the amino sugars found in the carbohydrate group linked to Asn78. The caprine amino acid sequence exhibits 88% identity with the bovine proteose-peptone component 3 sequence. However, when compared with the bovine sequence, the caprine sequence contains an insertion of a serine residue at position 25. PMID- 9749376 TI - Effects of controlled heat stress on ovarian function of dairy cattle. 1. Lactating cows. AB - The objective of this experiment was to determine the effects of controlled heat stress on ovarian function of lactating dairy cows. Estrus was synchronized (estrus = d 0), and cows were randomly assigned to either heat stress (n = 11; 29 degrees C, 60% relative humidity) or thermoneutral (n = 11; 19 degrees C, 60% relative humidity) treatment. For cows undergoing heat stress, ambient temperature (19 degrees C) was increased from d 11 to 13 of the estrous cycle (3.3 degrees C/d increase) and remained at 29 degrees C until d 21. Beginning on d 11, the growth and regression of ovarian follicles and corpora lutea were measured by using ultrasonography. Blood was collected daily by coccygeal venipuncture for measurement of serum concentrations of progesterone and estradiol. The second wave dominant follicle was more likely to ovulate in cows in the thermoneutral treatment than in cows undergoing heat stress (91 vs. 18% ovulation, respectively). Patterns of follicular growth in cows under-going heat stress were associated with decreased serum estradiol from d 11 to 21 and on the day of luteolysis. The average day of luteolysis was delayed by 9 d in heat stressed cows. Conclusions were that follicular growth and development and luteolytic mechanisms were compromised in heat-stressed cows; as a result, luteolysis was delayed, and second wave dominant follicles did not ovulate. PMID- 9749377 TI - Effects of controlled heat stress on ovarian function of dairy cattle. 2. Heifers. AB - The objective of this experiment was to determine the effects of controlled heat stress on ovarian function of dairy heifers. Estrus was synchronized in Holstein heifers (estrus = d 0), and heifers then were randomly assigned to either heat stress (n = 10; 33 degrees C, 60% relative humidity) or thermoneutral (n = 11; 21 degrees C, 60% relative humidity) treatment. For heat-stressed heifers, ambient temperature was increased from thermoneutrality to heat stress (33 degrees C) between d 9 and 14 (2.4 degrees C/d increase) after the synchronized estrus and remained between 31 and 33.5 degrees C until d 22. From d 11 to 21, the growth and regression of ovarian follicles and corpora lutea were measured by using ultrasonography, and blood was collected daily for serum progesterone and estradiol analyses. The second wave dominant follicle was larger for the heifers in the thermoneutral environment than for heat-stressed heifers, and ovulation of the second wave dominant follicle occurred in 9 of 11 thermoneutral heifers. For 6 of 10 heat-stressed heifers, the second wave dominant follicle regressed and was replaced by an ovulatory third wave dominant follicle. Smaller follicular size in heat stressed heifers was associated with decreased serum estradiol concentrations between d 11 and 21. Serum concentrations of progesterone during the luteal phase were similar, but luteolysis was delayed in heat-stressed heifers compared with onset in heifers in the thermoneutral treatment. Conclusions were that heat stress inhibited the growth and function of the dominant follicle so that most of the heat-stressed heifers had three follicular waves and a delay in corpus luteum regression. PMID- 9749379 TI - An electronic nose to detect changes in perineal odors associated with estrus in the cow. AB - Changes in perineal odor as estrus is approached could form the basis of a new method of estrus detection. Perineal odor of cyclic cows was monitored. Estrus was identified using ovarian ultrasound, behavioral observations, and plasma assay for progesterone and estradiol. Samples were taken from the dorsal lateral perineal (perivulval) area using cotton bud swabs and presented to an electronic nose. Twelve conducting polymer sensors were used to quantify odor in terms of a change in sensor resistance. Preliminary data (Experiment 1) indicate that the odor signals between the luteal phase and estrus could be distinguished for a group of five cows. In Experiment 2, samples were obtained daily from eight cows during the midluteal phase and from d -2 to d 8 of the cycle (d 0 = day of estrus, induced using cloprostenol). Seven of the eight cows cycled normally. Of the 12 sensors, 7 showed a significant change in resistance that was dependent on the day of the estrous cycle. Basal values were those taken in the luteal phase; values peaked on d -1, rose transiently on d 3, and returned to baseline on d 5 to 6. This pattern is strongly correlated with plasma estradiol concentration. The use of artificial olfaction could enable more accurate detection of estrus and has the potential to increase fertility in cows. PMID- 9749378 TI - Effect of time of artificial insemination on pregnancy rates, calving rates, pregnancy loss, and gender ratio after synchronization of ovulation in lactating dairy cows. AB - In order to assess the optimal time of artificial insemination (AI) in relation to ovulation, lactating dairy cows (n = 732) from herds with rolling herd averages of 9980 to 11,800 kg from three milkings per day were randomly assigned to five groups by stage of lactation and parity. Ovulation was synchronized by administration of GnRH followed 7 d later with PGF2 alpha followed 2 d later with a second treatment with GnRH. Cows were inseminated at 0, 8, 16, 24, or 32 h after the second injection of GnRH (ovulation occurs between 24 and 32 h after GnRH). Pregnancy diagnoses were performed by ultrasound at 25 to 35 d post-AI. Pregnancy rates per AI were similar for the groups inseminated at 0, 8, 16, and 24 h and lower for the group inseminated at 32 h. A significant quadratic effect of treatment suggests that the middle time periods (8, 16, and 24 h) may produce the greatest pregnancy rate per AI. However, the group inseminated at 0 h had lowest pregnancy loss, and the group inseminated at 32 h tended to have the greatest pregnancy loss compared with that of the other groups. The calving rate was similar between the groups inseminated at 0, 8, 16, and 24 h and lower in the group inseminated at 32 h. The time of AI also appeared to affect gender of calf: cows bred at 0 and 32 h having a higher percentage of female offspring. In conclusion, there appears to be substantial flexibility in the time of AI after the second injection of GnRH, and lower reproductive rates were observed only when AI was after the time of ovulation. PMID- 9749380 TI - Immunization of cows with ferric enterobactin receptor from coliform bacteria. AB - The serum and milk immunoglobulin (Ig) G responses of lactating dairy cows were determined following immunization with ferric enterobactin receptor FepA. Escherichia coli 471 was cultured in iron-depleted medium, and outer membrane proteins were extracted by 2% N-lauroylsarcosine sodium salt and 2% Triton X-100. The FepA was isolated from the outer membrane proteins by ion-exchange chromatography. Twenty cows were assigned to four treatment groups of 5 cows blocked by breed and days in milk. Treatment groups were vaccinated with 100 micrograms of FepA, 500 micrograms of FepA, Escherichia coli J5 bacterin, or sterile phosphate-buffered saline. Primary immunization was at approximately 200 d in milk, and booster immunizations were given 14 and 28 d later. Serum and whey IgG titers to FepA in cows vaccinated with FepA were significantly higher than those from cows vaccinated with either E. coli J5 bacterin or phosphate-buffered saline. Serum and whey IgG titers to FepA were elevated by 14 d in cows vaccinated with FepA. Significant differences were not observed between doses of FepA. The degree of cross-reactivity of purified IgG from cows vaccinated with FepA to E. coli and Klebsiella pneumoniae isolates was significantly higher than that to a control isolate that lacked FepA production. Immunization with FepA elicited an immunological response in serum and milk. PMID- 9749381 TI - Influence of route of vaccine administration against experimental intramammary infection caused by Escherichia coli. AB - The route of immunization of a commercially available Escherichia coli J5 bacterin was investigated. Jersey cows were randomly assigned to one of three treatment groups: 1) unvaccinated (control), 2) vaccinated subcutaneously in the neck, and 3) vaccinated in the area of the supramammary lymph node. Cows were vaccinated at drying off and at 2 wk prior to anticipated calving. Two quarters of each cow were challenged with approximately 60 cfu of E. coli at 14 d postcalving. Route of immunization in the neck or the area of the supramammary lymph node did not influence severity of coliform mastitis. However, the mean number of colony-forming units of E. coli recovered from challenged quarters was significantly lower for vaccinated cows than for control cows at 24 h postchallenge. A quicker milk yield recovery following intramammary challenge was also observed for vaccinated cows. Serum immunoglobulin (Ig) G, IgG1, and IgG2 and whey IgG1 and IgG2 antibody titers against E. coli J5 whole-cell antigens were significantly enhanced in vaccinated cows. Somatic cell counts in milk from challenged quarters and rectal temperatures following intramammary challenge were not different for cows across treatment groups. Immunization did not prevent intramammary infection. PMID- 9749382 TI - Effects of resistance to milk flow and the provision of hay on nonnutritive sucking by dairy calves. AB - This study examined effects of resistance to milk flow and the provision of hay on the duration of nutritive sucking and subsequent nonnutritive sucking by dairy calves. In a series of four experiments, 12 male Holstein calves were individually fed milk from an artificial teat. Resistance to milk flow was varied by adjusting the orifice size within the milk supply tube. Using a Latin square design, each calf was fed the same quantity of milk using four orifice sizes (one per day for 4 consecutive d). The duration of nutritive sucking (time required to finish the milk meal) was longer when calves were fed from the smallest orifice size (0.16-cm diameter) than when calves were fed from the largest orifice size (0.55-cm diameter). Calves compensated for resistance to milk flow in an attempt to maintain milk intake; however, changes in sucking rate alone were probably not responsible for observed differences in the rate of milk consumption from different orifice sizes. The duration of nonnutritive sucking (sucking of the teat following the milk meal) was significantly reduced when calves were fed from the smallest orifice compared with that when calves were fed from the largest orifice. Nonnutritive sucking was reduced but not eliminated even with the longest meal duration. The provision of hay to calves following the meal also significantly reduced the amount of nonnutritive sucking. Reduction in flow rate when calves drink milk through a teat and the provision of hay after the meal can reduce the incidence of nonnutritive sucking following the meal and may help to limit cross-sucking in group housing systems. PMID- 9749383 TI - Effects of high calcium intake on fat digestion and bile acid excretion in feces of veal calves. AB - We tested the hypothesis that apparent digestibility of fat by veal calves is determined by the participation of bile acids in the process of fat absorption and is, therefore, negatively associated with bile acid excretion in feces. Veal calves were fed milk replacers that contained whey protein and either a low (5.2 g of calcium/kg of air-dried diet) or high (12.4 g of calcium/kg of air-dried diet) concentration of calcium. The high calcium milk replacer contained extra calcium in the form of calcium formate. Final body weight was not significantly influenced by diet after the milk replacers had been fed for 27 wk. Feces were quantitatively collected during wk 23 of the trial. The high calcium milk replacer reduced apparent fat digestibility by 5.6 percentage units and increased bile acid excretion in feces by 90% compared with the low calcium milk replacer. The extra calcium intake decreased apparent absorption of magnesium and phosphorus. We proposed that a high intake of calcium by veal calves would increase the amount of insoluble calcium, magnesium, and phosphate complexes in the intestinal lumen, which, because of the binding of bile acids, would exclude bile acids from the process of fat digestion and inhibit reabsorption of bile acids. As a result, fat digestion is impaired, and bile acid excretion in feces is enhanced. The complex formation in the ileal lumen also explains why high calcium intake reduces magnesium and phosphorus absorption in veal calves. PMID- 9749384 TI - Influence of genotype and ensiling of corn grain on in situ degradation of starch in the rumen. AB - This trial was conducted to determine the influence of genotype and ensiling of corn grain on the rate and extent of ruminal starch degradation. Two cultivars of corn that differed in texture of the endosperm, dent (Zea mays ssp. indentata) or flint (Zea mays ssp. indentura) were harvested at 30% whole-plant dry matter (DM). After separation from stover and cob, the kernels were coarsely chopped and ensiled or not ensiled. Grains were oven-dried at 40 degrees C and either ground through a 3-mm sieve or left unground. Ruminal DM and starch degradabilities were determined using the in situ technique. The proportion of starch lost through the pores of the bag without degradation was also determined. Mean ruminal DM and starch degradabilities were higher for ground grains than for chopped grains, which could be related to the proportion of DM and starch lost through the pores of the bag. For unensiled, chopped grain, ruminal starch degradability was higher for dent corn than for flint corn (72.3% vs. 61.6%). The ensiling process increased ruminal starch degradability, averaging 5.8 percentage units. The difference in ruminal starch degradability between dent corn and flint corn remained constant whether the corn was unensiled or ensiled (10.7 vs. 11.6 percentage units). PMID- 9749385 TI - Biogenic amines in silage, apparent postruminal passage, and the relationship between biogenic amines and digestive function and intake by steers. AB - A 4 x 4 Latin square experiment was conducted to examine abomasal passage of biogenic amines in steers fed silage and their related effects on intake, digestibility, and digestive function. Thirty percent of the dry matter (DM) in the diets consisted of alfalfa forage, which was fed as either hay or silage. The DM from alfalfa silage DM was substituted at 0, 33, 67, and 100% for DM from alfalfa hay and was fed to four ruminally and abomasally cannulated steers. The roughage component of the diet constituted 50% of the DM and consisted of 60% alfalfa silage or hay and 40% tropical corn silage. The concentrate was composed mainly of ground corn. The concentrations of putrescine and cadaverine in abomasal digesta increased as alfalfa silage in the diet increased. Abomasal recovery of biogenic amines, a product of their concentration in abomasal digesta and the passage of DM through the abomasum, was negatively correlated with intake. Abomasal recovery of most amines was 5 to 20% of intake. Abomasal recovery of cadaverine was correlated with depressed intake. Total DM intake was reduced 8.3 to 25.8% as the proportion of alfalfa silage in the diet increased. Frequency of reticular contractions, intake, ruminal DM digestibility, ruminal outflow, volatile fatty acids, and total tract DM digestibility decreased in steers fed diets that contained more alfalfa silage. Ruminal fluid pH and NH3 concentration increased in steers fed more alfalfa silage; however, mass and the DM percentage of ruminal contents decreased linearly. Postprandial insulin concentrations were quadratically related to the proportion of alfalfa hay or silage in the diet. Intraruminal metabolism of biogenic amines is extensive based on the relatively low quantities recovered in abomasal digesta; however, the amounts recovered in abomasal digesta were related to intake depression and associated physiological effects. PMID- 9749386 TI - Supplementation of rumen-undegradable protein to the diets of early lactation Holstein cows on grass pasture. AB - The objectives were to evaluate supplements differing in rumen-undegradable protein (RUP) on the performance of early lactation cows on grass pasture. Twenty four Holstein cows averaging 68 d of lactation and 39.8 kg/d of milk were rotationally grazed on predominantly Dactylis glomerata pasture for 8 wk. Cows were blocked according to parity, milk yield, and days of lactation and were randomly assigned to a grain mixture with a low or high RUP content. Pasture and the low and high RUP grain mixtures averaged 25.6, 14.7, and 13.7% crude protein and 4.0, 7.0, and 8.4% RUP, respectively (dry matter basis). Grain was fed twice daily at 1 kg/4 kg of milk, pasture provided all forage in the diet, and grain consumption was similar (8.9 kg/d per cow) for cows on both treatments. Total dry matter intakes estimated using chromic oxide were 19.9 and 20.9 kg/d for cows fed low and high RUP grain mixtures. Milk yields did not differ between treatments; means were 34.2 and 35.5 kg/d for cows fed low and high RUP grain mixtures. Multiparous cows tended to yield more milk (36.2 vs. 34.5 kg/d) and milk protein (1.06 vs. 0.98 kg/d) when fed the high RUP grain mixture. Concentrations of plasma urea N and nonesterified fatty acids were unaffected by treatment and averaged 18.7 mg/dl and 307 microeq/L, respectively. Results indicated that a supplemental grain mixture with a high RUP content did not alter milk yield of high yielding cows when pasture was the sole forage; however, milk protein yield tended to be greater for multiparous cows fed the high RUP grain mixture. PMID- 9749387 TI - Effect of ruminally protected amino acids on milk yield and composition of Jersey cows fed whole cottonseed. AB - The objectives of this experiment were to determine whether ruminally protected amino acids (AA) increased milk protein when this content was depressed by the addition of whole cottonseeds in the diets of early lactation Jersey cows. Treatments were 1) a control diet, 2) a diet containing whole cottonseed, and 3) a diet containing whole cottonseed and ruminally protected lysine and methionine. Cows were assigned to treatments at a mean of 7 d postpartum and remained on the experiment for 18 wk. Dry matter intake and yields of milk, milk fat, fat corrected milk, and energy-corrected milk were not affected by treatment. Milk fat content tended to decrease for cows fed diets containing whole cottonseed. However, the percentages of milk protein, total N, and casein N were depressed by the addition of whole cottonseed and were increased by the addition of ruminally protected AA. Plasma concentrations of methionine, but not lysine, were increased when ruminally protected AA were fed, suggesting that lysine was the most limiting. PMID- 9749388 TI - Starch supplementation of grass harvested at two stages of maturity prior to ensiling: intake, digestion, and degradability by dairy cows. AB - The effects of the maturity of grass prior to ensiling and the supplementation of starch to grass silage on apparent digestibility, degradability, rumen content, and feed intake by dairy cows were investigated using a Latin square design. Treatments were silages from early or late cut grass with or without 4 kg of supplemental flaked corn starch. The silage from early cut grass contained more N and sugars than did the silage from late cut grass but was lower in neutral detergent fiber (NDF). Degradation characteristics were not different between the two silages. Apparent digestibilities of dry matter (DM), organic matter (OM), and N in the silage from late cut grass were lower than those in the silage from early cut grass, but NDF digestibility was not affected. Starch supplementation increased the lag phase for DM and OM in both silages, but the rate of degradation was decreased for NDF. Starch supplementation did not influence digestibilities of DM and OM in silage from early cut grass but decreased the digestibilities of DM and OM in silage from late cut grass. Crude protein and NDF digestibilities were decreased for silages from early and late cut grass. Starch supplementation increased NDF in the rumen of cows fed the silage from early cut grass, but NDF was not affected by starch supplementation for cows fed the silage from late cut grass. Rumen-degradable starch negatively influences degradability and apparent OM digestibility; the extent of the decrease is related to the maturity of the NDF. PMID- 9749389 TI - Feed intake relative to stage of lactation for dairy cows consuming total mixed diets with a high or low ratio of concentrate to forage. AB - This experiment examined the effect of feed quality on the relationship between intake and stage of lactation in dairy cows. Two total mixed diets composed of grass silage and concentrate were formulated. The high concentrate total mixed diet was designed to meet energy requirements, and the low concentrate total mixed diet was designed to limit intake. Twenty-four Holstein-Friesian cows were offered the total mixed diets in a full 2 x 2 change-over design with control treatments. The changeover was at 153 d in milk (DIM). For the statistical analyses, two periods of 13 wk, one period before and one period after the changeover, were used. Dry matter intake (DMI), milk yield, body weight, and body condition score were significantly greater for cows fed the high concentrate total mixed diet than for cows fed the low concentrate total mixed diet. Significant interactions between total mixed diet and period were observed for DMI and milk yield. However, no significant residual effects of changing from one total mixed diet to the other were observed. The interactions were due to substantially different slopes of DMI and milk yield relative to DIM for cows fed the two different total mixed diets. For cows fed the low concentrate total mixed diet, there was no effect of stage of lactation on DMI; the slope was 0. For cows fed the high concentrate total mixed diet, there was a significant decline in DMI as lactation progressed. PMID- 9749390 TI - Long-term effects of acetate and propionate on voluntary feed intake by midlactation cows. AB - Our objective was to separate the effects of physical fill and acetate production in the regulation of voluntary feed intake. Eight ruminally fistulated Holstein cows in midlactation were fed a low forage diet or a high forage diet with or without continuous ruminal infusion of buffered acetate or propionate in a duplicated 4 x 4 Latin square with 21-d periods. Cows consumed about 3.5% of body weight as dry matter, and voluntary dry matter intake (DMI) was approximately 6% greater when cows were fed the low forage diet than when cows were fed the high forage diet. Infusion of 7.1 Mcal of net energy for lactation as acetate or propionate resulted in a reduction in DMI relative to the DMI when the high forage diet was fed alone; propionate infusion reduced intake more than did acetate infusion. Consumption of neutral detergent fiber was approximately 1.19 and 1.25% of body weight when cows were fed the low and high forage diets, respectively. Milk production was approximately 35 kg/d regardless of the diet fed, but an increase in milk fat production by cows receiving the acetate or propionate infusion resulted in an increase in fat-corrected milk. Neither neutral detergent fiber fill nor a threshold for acetate utilization appeared to limit DMI. PMID- 9749391 TI - Inhibition of sulfate reduction to sulfide by 9,10-anthraquinone in in vitro ruminal fermentations. AB - We studied the effects of sulfur and 9,10-anthraquinone on in vitro ruminal fermentation and production of hydrogen sulfide. A complete, pelleted diet containing 26.8% acid detergent fiber, 15.9% crude protein, and 0.25 to 0.29% sulfur was used as the basal substrate. Fermentations were conducted using the basal substrate and various sulfur additions (elemental sulfur, thiosulfate, calcium sulfate, and sodium sulfate) with or without varying amounts of 9,10 anthraquinone. An increase in the sulfur content of the substrate to > 1.0% with various sources of sulfur had minimal effects on concentrations of volatile fatty acids, but the production of hydrogen sulfide increased. High amounts of 9,10 anthraquinone (10 and 25 ppm of fluid) decreased the molar proportion of acetate and decreased the production of methane and hydrogen sulfide. However, 9,10 anthraquinone increased the molar proportions of propionate and butyrate. Approximately 70% of 9,10-anthraquinone was recovered after 24 h of in vitro ruminal fermentation. These findings suggest that 9,10-anthraquinone has the potential to reduce the production of methane and hydrogen sulfide in ruminal fermentations. PMID- 9749392 TI - Use of multinational data to improve national evaluations of Holstein bulls. AB - The usefulness of multinational data for the improvement of national estimates of genetic merit of Holstein bulls was assessed. For 222 bulls, combined US-Canadian evaluations and evaluations from the US only from January 1993 for milk, fat, and protein yields were compared with their US only evaluations from August 1997. The correlations between the 1993 and 1997 evaluations and the standard deviations of differences in evaluations from added data favored the evaluations from the US only because of a part-whole relationship; often 1997 data were largely from US only data from 1993. However, the results for 35 bulls with reliability increases of > 5% indicated that combining US and Canadian evaluations improved the prediction of future evaluations. The value of foreign data also was assessed from national and international evaluations on the scales of Canada, Germany, and the US. The changes from 1996 national evaluations to either 1996 international evaluations or 1997 national evaluations were compared to determine whether adding international data at the earlier time could provide a useful prediction of subsequent change in national evaluations. Although the degree of agreement among differences from added national and international data varied, international evaluations did provide useful information beyond the more limited national data available at the same time. PMID- 9749393 TI - Genetic parameters of health disorders, and relationships with 305-day milk yield and conformation traits of registered Holstein cows. AB - A total of 4368 first lactation records for Holstein cows from 30 herds was used to estimate genetic parameters for yield, conformation traits, and the binary coded disease traits of udder edema, milk fever, retained placenta, metritis, displaced abomasum, ketosis, cystic ovary, mastitis, and lameness. Data on health, parentage, and yield came from an on-farm program for record keeping and management. Test day production data were obtained from British Columbia DHI. Type classification data were received from the Holstein Association of Canada. Heritabilities of disease traits were low ranging from 0 to 0.05. Exceptions were lameness (0.16) and ketosis (0.39). Correlations of disease traits with 305-d milk yield and of selected type traits with retained placenta, displaced abomasum, mastitis, and lameness were estimated. Phenotypic correlations did not substantially differ from 0 except for the correlation between lameness and rear leg set (0.37). Genetic correlations between disease traits and milk yield were mostly positive (0.02 to 0.44). Only retained placenta had a negative genetic correlation with milk yield (-0.28). Genetic correlations ranged from 0 to 0.37 between udder conformation traits and mastitis, from -0.38 to 0.09 between leg conformation traits and lameness, and from -0.11 to 0.38 between rump conformation and retained placenta. The results suggest that selection based solely on yield may increase the incidence of disease. Selection on conformation traits can help reduce the incidence of disease, although genetic correlations are low. PMID- 9749394 TI - Influence of production circumstances and economic evaluation criteria on economic comparison of breeds and breed crosses. AB - The ranking of genotypes (i.e., breeds and breed crosses) for economic performance depends on the production circumstances of the herd and the criteria for economic evaluation. In this study, the effects of evaluation criteria and production circumstance are quantified using data from the literature on six genotypes. The economic evaluation criteria measured at herd level included lifetime profit and lifetime profit expressed per day of calving interval, per day of productive lifetime, per day of total herd life, per unit of milk, and per unit of feed energy. Four production circumstances were studied; these circumstances included constraints on the output of milk and on herd use of concentrates, roughages, and both concentrates and roughages. Profit was determined based on the sale of milk, calves, cull cows, and heifers and the expenses incurred for concentrates and roughages, animal deaths, milking time, interest, and other inputs. Results indicate that production circumstances and evaluation criteria largely influence the ranking of genotypes and, therefore, the outcome of economic comparisons. The genotype that ranks the best under a certain criteria and circumstance ranks differently when these situations are altered. In economic comparisons of genotypes aimed at offering recommendations for implementation in smallholder dairy production systems in the tropics, the choice of the evaluation criteria should be determined by the limitation or constraint that characterizes the area where the genotypes are to produce or are currently producing. For example, for situations in which feed availability is limiting, genotypes should be ranked on total profit per unit of feed energy. PMID- 9749395 TI - Partial budget of the discounted annual benefit of mastitis control strategies. AB - The objective of this study was to rank the benefits associated with various mastitis control strategies in simulated herds with intramammary infections caused by Streptococcus agalactiae, Streptococcus spp. other than Strep. agalactiae, Staphylococcus aureus, coagulase-negative staphylococci, and Escherichia coli. The control strategies tested were prevention, vaccination for E. coli, lactation therapy, and dry cow antibiotic therapy. Partial budgets were based on changes caused by mastitis control strategies from the mean values for milk, fat, and protein yields of the control herd and the number of cows that were culled under a fixed mastitis culling criterion. Each annual benefit (dollars per cow per year) of a mastitis control strategy was compared with the revenue for the control herd and was calculated under two different milk pricing plans (3.5% milk fat and multiple-component pricing), three net replacement costs, and three prevalences of pathogen-specific intramammary infection. Twenty replicates of each control strategy were run with SIMMAST (a dynamic discrete event stochastic simulation model) for 5 simulated yr. Rankings of discounted annual benefits differed only slightly according to milk pricing plans within a pathogen group but differed among the pathogen groups. Differences in net replacement costs for cows culled because of mastitis did not change the ranking of control strategies within a pathogen group. Both prevention and dry cow therapy were important mastitis control strategies. For herds primarily infected with environmental pathogens, strategies that included vaccination for mastitis caused by E. coli dominated strategies that did not include vaccination against this microorganism. PMID- 9749396 TI - Germicidal activity of a chlorous acid-chlorine dioxide teat dip and a sodium chlorite teat dip during experimental challenge with Staphylococcus aureus and Streptococcus agalactiae. AB - Three postmilking teat dips were tested for efficacy against Staphylococcus aureus and Streptococcus agalactiae in two separate studies using experimental challenge procedures that were recommended by the National Mastitis Council. The first study evaluated a barrier teat dip product containing chlorous acid chlorine dioxide as the germicidal agent, and the second study evaluated a sodium chlorite product with a barrier component as well as a sodium chlorite product without a barrier component. The chlorous acid-chlorine dioxide teat dip reduced new intramammary infections (IMI) caused by Staph. aureus by 91.5% and reduced new IMI caused by Strep. agalactiae by 71.7%. The barrier dip containing sodium chlorite reduced new IMI caused by Staph. aureus and Strep. agalactiae by 41.0 and 0%, respectively. The nonbarrier dip containing sodium chlorite reduced new IMI caused by Staph. aureus by 65.6% and reduced new IMI caused by Strep. agalactiae by 39.1%. Teat skin and teat end conditions were evaluated before and after the second study; no deleterious effects among dipped quarters compared with control quarters were noted for the two sodium chlorite products. PMID- 9749397 TI - A summary of the reasons why farmers cull cows. AB - A study was conducted to determine why cows are culled, whether cows are culled for multiple reasons, and whether farm characteristics can help explain why cows are culled. The primary reasons for culling were reproduction (i.e., failure to conceive), mastitis, and low production. For 35% of all cows that were culled, a secondary reason for culling was assigned by the farmer, and, for 11% of all cows that were culled, a tertiary reason was recorded. Using a weighting factor of 5:3:1 (primary:secondary:tertiary reasons for culling), a culling score was computed for each reason within a herd. Reproduction was the primary reason for culling, production was second, and mastitis was third. Much variation existed as to why dairy farmers culled cows. Computed mean culling scores suggested that culling for mastitis was lower in high producing Holstein herds, and culling for abortion was higher in high producing Holstein herds. Culling for reproduction was higher in high producing Holstein herds, and culling for production was lower in high producing Holstein herds. Culling for mastitis and production were significantly lower in high producing Holstein herds than in non-Holstein herds. A system that allows the documentation of multiple reasons for culling and the computation of a composite score appears to be reasonable for the assessment of culling management. PMID- 9749398 TI - Breed group effects on milk production of Brazilian crossbred dairy cows. AB - Lactation records (n = 2362) of 1402 crossbred cows in 22 cooperating dairy herds in southeastern Brazil were evaluated. Cows were mixtures of Zebu (Gir, Guzera, and unknown) and European breeding (mostly Holstein). Lactation milk yields were expressed as total, 3050-d, or deviated 305-d yields, either adjusted or unadjusted for days in milk (DIM). Mean DIM was 280. Arithmetic means unadjusted for DIM were 1942, 1666, and 5 kg per record. Milk yields of daughters from sires of 6/8 and 7/8 European breeds were higher than yields of daughters from sires of 5/8 European breeds when data were either adjusted or unadjusted for DIM. The differences associated with breed group of sire were only slightly reduced when records were adjusted for DIM. There was no evidence of a decline in milk yield as the fraction of European breeding of the sire increased from 6/8 to 7/8. For a given breed group of sire, whether the grandsire was purebred or crossbred had no detectable effect. These results should be useful in determining strategies for crossbreeding of dairy cows in tropical areas, particularly when crossbred sires are used. PMID- 9749399 TI - Revised guidelines for the control of methicillin-resistant Staphylococcus aureus infection in hospitals. British Society for Antimicrobial Chemotherapy, Hospital Infection Society and the Infection Control Nurses Association. PMID- 9749400 TI - A hospital outbreak of extended-spectrum beta-lactamase-producing Klebsiella pneumoniae investigated by RAPD typing and analysis of the genetics and mechanisms of resistance. AB - Between July and September 1997 a ceftazidime- and aminoglycoside-resistant strain of Klebsiella pneumoniae infected or colonized seven patients on three paediatric wards at Guy's Hospital in London. The patients were mostly neonates or infants recovering from cardiac surgery for congenital defects. The organism was probably introduced by an asymptomatic patient from Greece and the subsequent outbreak could largely be explained by person-to-person spread on individual wards and frequent transfers of patients between wards. The outbreak was controlled by patient isolation and attention to handwashing, and there were no fatalities. The organisms were non-typeable by serology but had a characteristic RAPD profile. They produced the extended spectrum beta-lactamase SHV-5 and the aminoglycoside-modifying enzymes AAC(6') + probably AAC(3)II, encoded on a conjugative plasmid of approximately 160 kb. Two other patients had multi resistant klebsiellas, one of them an SHV-5 producer and one a TEM-5 producer, but these could be distinguished from each other and from the outbreak strain by serological and RAPD typing and by the genetics and mechanisms of their resistances. Three other multi-resistant enterobacteria were isolated during the outbreak: an Escherichia coli that had acquired the 160 kb resistance plasmid from the epidemic klebsiella, a Citrobacter isolated from one of the patients with the klebsiella but which did not produce SHV-5, and a TEM-5-producing Enterobacter. This outbreak illustrates the importance of screening patients from high-risk areas for multiply-resistant organisms on admission, and the value of bacterial typing and analysis of resistance mechanisms to define the epidemiology of hospital infection. PMID- 9749401 TI - Outbreak of nosocomial urinary tract infections due to Pseudomonas aeruginosa in a paediatric surgical unit associated with tap-water contamination. AB - An outbreak of 14 cases of urinary tract infections by Pseudomonas aeruginosa, including six symptomatic infections, occurred from September to November 1994 in a paediatric surgical unit. During the outbreak, urine samples from patients and multiple samples from the environment of patients were tested for the presence of P. aeruginosa. Bacterial isolates were studied by pulsed-field gel electrophoresis. Genotypic analysis showed that most of the isolates from children were different. Multiple P. aeruginosa isolates were also found in the tap water, as the only putative source of contamination. Two of these isolates were identified in two infected patients, indicating possible direct contamination of patients via tap water and this was related to the distal colonization of faucets. Bacteria were eradicated from tap water by replacement of taps. The cluster of cases of P. aeruginosa urinary infection was, therefore, related to multiple contaminations through tap water. These results illustrate an unexpected risk of nosocomial infection and emphasizes the importance of checking tap water to prevent bacterial contamination through handwashing in contaminated water. PMID- 9749403 TI - Closed tracheal suctioning systems and infection control in the intensive care unit. AB - Closed tracheal suction catheters offer a number of microbiological advantages over the conventional single-use suction catheters. Intensive care staff, however, have experienced difficulties such as pooling of the catheter irrigation saline within the connectors, and hand contamination from condensate which escapes via the irrigation port. Using a descriptive survey design we quantified how frequently these problems occurred. Over an eight-week period, staff completed 923 survey forms. Hand contamination from condensate was reported in 61% of responses. Rinsing the catheter after use was ineffective in 39% of responses, and 70% reported pooling of the saline in the swivel and ventilator connectors. Forty-five percent of responses reported ineffective secretion removal. The infection implications for clinical practice are discussed. PMID- 9749402 TI - Neonatal infections with Pseudomonas aeruginosa associated with a water-bath used to thaw fresh frozen plasma. AB - In our 15-bed neonatal intensive care unit (NICU), four new-borns were found to be colonized or infected with Pseudomonas aeruginosa within a period of one week. To identify the outbreak source, three independent studies were performed: epidemiological investigation, environmental surveillance and genotypic typing of isolates. Although epidemiological investigation by a case-control study revealed no conclusive results, the transfusion of fresh frozen plasma (FFP) and human albumin (HA) appeared to be the factor with highest risk. Environmental surveillance and random amplification of polymorphic DNA (RAPD) of isolates identified a water-bath used to warm FFP and HA as the likely reservoir for the outbreak. Further spread of the organism did not occur after elimination of this water-bath from the NICU. RAPD identified in addition an isolate from an infant hospitalized in the NICU five months before the outbreak with a pattern matching the one of the outbreak cluster. PMID- 9749404 TI - Role of antimicrobial-impregnated polymer and Teflon in the prevention of biliary stent blockage. AB - Biliary stent blockage and microbial colonization is a common complication associated with polyurethane stents used for the relief of bile-duct obstruction caused by benign or malignant disease. In an attempt to overcome this problem the application of a 'Teflon' (polytetrafluoroethylene) stent and an antimicrobial benzalkonium chloride (BZC) impregnated polymer were investigated. The effects of these materials on microbial colonization were compared to a polyurethane stent in vitro in broth or bile. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of BZC for three commonly isolated biliary stent pathogens, Staphylococcus epidermidis, Enterococcus faecium and Enterobacter cloacae were also determined. All the isolates were sensitive to BZC. The growth kinetics of the three organisms in broth and in human pooled bile were similar. Adherence to the BZC impregnated polymer was significantly reduced as compared to the polyurethane and Teflon stents (P < 0.05) in nutrient broth. In bile, fewer organisms attached to the Teflon as compared with the polyurethane stent (P < 0.05) for all organisms. For two of the three test organisms there was less bacterial adherence to the Teflon than to the BZC impregnated polymer. The Teflon and antimicrobial stent materials studied may prevent biliary stent blockage resulting from microbial colonization. PMID- 9749405 TI - Infection control policies for anaesthetic equipment must be comprehensive. PMID- 9749406 TI - Late eocene sivaladapid primate from Guangxi Zhuang Autonomous Region, People's Republic of China. AB - A new genus and species of Sivaladapidae is described from the late Eocene Gongkang Formation, Yongle Basin, western Guangxi Zhuang Autonomous Region, southern China. Guangxilemur tongi, new genus and species, shows a combination of traits that occur separately in earlier and more primitive Asian adapiforms (Hoanghonius and Rencunius) and in Miocene sivaladapines (Sivaladapis and Sinoadapis). Phylogenetic analysis of dental characters suggests that Guangxilemur is closely related to the Miocene sivaladapine clade. Miocene sivaladapines were the latest surviving members of a broad radiation of Eocene adapiforms in Asia that included Hoanghonius, Rencunius, and Wailekia in addition to Guangxilemur. European Periconodon may also be specially related to this primarily Asian clade, but current anatomical data are insufficient to test this possibility adequately. Sivaladapine adapiforms and tarsiid tarsiiforms maintained relictual distributions in southern and/or southeastern Asia far beyond the extirpation of their closest relatives on other Holarctic continents near the Eocene-Oligocene boundary. This temporal persistence was mediated by Asian paleogeography, which allowed virtually continuous access to tropical refugia during a middle Cenozoic interval of climatic deterioration that coincided with the extinction of adapiforms and tarsiiforms in Europe and North America. PMID- 9749407 TI - Phylogenetic analysis of anthropoid relationships. AB - The relationships of anthropoids to other primates are currently debated, as are the relationships among early fossil anthropoids and crown anthropoids. To resolve these issues, data on 291 morphological characters were collected for 57 taxa of living and fossil primates and analyzed using PAUP and MacClade. The dental evidence provides weak support for the notion of an adapid origin for anthropoids, the cranial evidence supports the tarsier-anthropoid hypothesis, and the postcranial evidence supports a monophyletic Prosimii and a monophyletic Anthropoidea. Combining these data into a single data set produces almost universal support for a tarsier-anthropoid clade nested within omomyids. Eosimias and Afrotarsius are certainly members of this clade, and probably basal anthropoids, although the Shanghuang petrosal may not belong to Eosimias. The tree derived from the combined data set resembles the tree derived from the cranial data set rather than the larger dental data set. This may be attributable to relatively slower evolution in the cranial characters. The combined data set shows Anthropoidea to be monophyletic but the features traditionally held to be anthropoid synapomorphies are found to have evolved mosaically. Parapithecines are the sister taxon to crown anthropoids; qatraniines and oligopithecids are more distantly related sister taxa. There is support for a relationship of a Tarsius + Anthropoidea clade with either washakiines on Uintanius. These elements of tree topology remain fairly stable under different assumptions sets, but overall, tree topology is not robust. Previously divergent hypotheses regarding anthropoid relationships are attributable to the use of restricted data sets. This large data set enables the adapid-anthropoid hypothesis to be rejected, and unites Tarsius, Anthropoidea and Omomyiformes within a clade, Haplorhini. However, relationships among these three taxa cannot be convincingly resolved at present. PMID- 9749408 TI - Radiators are cool: a response to Braga & Boesch's published paper and reply. PMID- 9749409 TI - Hominoid phylogeny: inferences from a sub-terminal minisatellite analyzed by repeat expansion detection (RED) PMID- 9749410 TI - Concerning the three stage model of carcass processing at FLK Zinjanthropus: a reply to Capaldo. PMID- 9749413 TI - Combined cytology and colposcopy to screen for cervical cancer in pregnancy. AB - OBJECTIVE: To evaluate the accuracy of cytologic screening in pregnancy through routine colposcopy and to confirm the safety of conservative management of cervical intraepithelial neoplasia (CIN) in pregnancy. STUDY DESIGN: In total, 3,658 pregnant women, screened for cervical cancer with either cytology or colposcopy, were prospectively evaluated. Patients with abnormal findings underwent colposcopically directed biopsy and, in case of CIN, repeat cytology and colposcopy. Biopsy was repeated in case of suspected progression of the lesion. Suspected microinvasion was the only reason for diagnostic conization during pregnancy. After delivery, excisional treatment provided a final specimen from all patients. Diagnostic methods were compared. RESULTS: Comparison between cytology and colposcopy showed 97.1% concordance with a few false positives (2.5%) and false negatives (0.2%). Abnormal cytology and colposcopy, as compared with histology, showed similar concordances, but the risk of underestimation by cytology was significantly higher (P < .05). Initial and final histology of the 63 cases of CIN and microinvasive carcinoma showed 88.9% concordance. Progression of the lesion was not observed. CONCLUSION: These data do not justify combined use of cytology and colposcopy to improve screening for cervical cancer in pregnancy. Delayed treatment of CIN after delivery is safe. PMID- 9749414 TI - Can shoulder dystocia be predicted? Preconceptive and prenatal factors. AB - OBJECTIVE: To evaluate the predictability of shoulder dystocia using preconceptive and prenatal risk factors. STUDY DESIGN: Data from 1,622 term patients with prenatal care prior to 20 weeks who delivered single, vertex fetuses during a consecutive 12-month period were analyzed. Two groups were chosen. The first group was patients whose fetuses experienced shoulder dystocia during delivery (cases). The second group (controls) consisted of the remaining patients, whose fetuses had not experienced shoulder dystocia. The two groups were compared with regard to demographics and pregnancy characteristics. RESULTS: Factors not significantly different between the two groups included were obesity, multiparity, history of diabetes, short maternal stature, postdatism and advanced maternal age. The incidence of macrosomia was significantly higher (P < .001) in cases (35.4%) than in controls (4.8%). Other factors associated with shoulder dystocia were previous shoulder dystocia, concurrent diabetes, prior delivery of a fetus > 4,000 g and excessive weight gain during pregnancy. Many factors previously associated with shoulder dystocia were found to be nonsignificant in our study. CONCLUSION: Macrosomia appears to be the single important factor associated with shoulder dystocia which, even in the presence of significant risk factors, remains largely unpredictable. PMID- 9749412 TI - Update on vertical HIV transmission. AB - OBJECTIVE: To describe the factors that contribute to vertical transmission of human immunodeficiency virus (HIV) and review means of decreasing the risk of transmission. STUDY DESIGN: Medline search of the international English-language literature pertaining to HIV in pregnancy from 1989 to the present. Special emphasis was placed on articles published in the last three years related to vertical transmission as well as to antepartum, intrapartum and postpartum management to reduce transmission. RESULTS: High levels of maternal viral load and more advanced maternal disease are associated with a greater risk of vertical transmission of HIV. Antepartum and intrapartum maternal treatment with zidovudine and postpartum neonatal zidovudine treatment decreases the risk of transmission by two-thirds, at least in patients with earlier stages of the disease. Breast-feeding is a source of postpartum HIV transmission and may double the total transmission rate. CONCLUSION: Zidovudine should be used in pregnancy to decrease the viral load and reduce transmission of HIV to the fetus. Other antiviral agents should be used during pregnancy if indicated, although current information is lacking about their effects on the fetus and any potential benefits in decreasing vertical transmission of HIV. Breast-feeding should be avoided. PMID- 9749415 TI - Prenatal diagnosis of limb-body wall complex. AB - OBJECTIVE: To evaluate prenatal diagnosis of limb-body wall complex (LBWC) by ultrasonography in eight cases. STUDY DESIGN: The diagnosis was based on two of the following: exencephaly/encephalocele with facial clefts, thoracoschisis and/or abdominoschisis and limb defect. The ultrasonographic findings were compared with the autopsy findings in each case. RESULTS: The average weeks of gestation at which malformations were diagnosed by ultrasonography was 21.7 +/- 4.7 (mean +/- SD, n = 8). All eight fetuses were diagnosed as having characteristic abnormalities and six of them as having scoliosis by ultrasonography. Four of the eight were examined for maternal serum alpha fetoprotein (MSAFP); the levels exceeded 2.5 multiples of the mean according to the standard value at our hospital. Chromosomal analysis was performed for six cases and revealed that they were normal in karyotype. All eight cases showed abdominoschisis, scoliosis and abnormalities of the lower extremities. A single umbilical artery was present in seven cases (87.5%), and a short umbilical cord was present in seven (87.5%). CONCLUSION: Ultrasonographic detection of abdominoschisis, scoliosis abnormalities of the lower extremities, a single umbilical artery and a short umbilical cord is important for the prenatal diagnosis of LBWC. An extremely elevated level of MSAFP is also indicative of the complex. PMID- 9749416 TI - Differential binding of estradiol and testosterone to SHBG. Relation to circulating estradiol levels. AB - OBJECTIVE: Sex hormone-binding globulin (SHBG) binds testosterone (T) to a greater degree than it does estradiol (E2), acting as an amplifier of E2 action. However, it is not known whether the relative capacity of SHBG for E2 vs. T is altered by the hormonal milieu. We hypothesized that an increase in circulating E2 levels results in a compensatory increase in the relative binding capacity of SHBG for these hormones, dampening the E2 amplification effect in hyperestrogenic conditions. STUDY DESIGN: Retrospective. RESULTS: As expected, during hMG stimulation there was a significant increase in total and free E2 (28 to 1,986 pg/mL, P < .001; and 0.3 to 20.8 pg/mL, P < .001, respectively) and total T levels (40.3 vs. 78.3 ng/dL, P < .001) from basal to late stimulation. Free T levels increased, but the difference did not reach significance. The binding capacity of SHBG for both E2 and T increased in a proportional manner (980 +/- 340 vs. 1,434 +/- 449 nmol/L, P < .009; and 352 +/- 190 vs. 512 +/- 128 nmol/L, P < .02; respectively) since the ratio of SHBG binding to E2 and T was unchanged. Although the SHBG molar concentration appeared increased, the difference did not reach significance (821 +/- 542 to 1,099 +/- 254 nmol/L). CONCLUSION: A short term, although profound, increase in circulating E2 does not seem to be associated with an increase in the relative binding capacity of the carrier protein for either E2 or T, although an overall increase in binding for both steroids was observed. It is possible that longer periods of exposure to E2 may be necessary to demonstrate a change in the differential binding of this carrier protein with an alteration in the hormonal milieu. PMID- 9749418 TI - Unengaged vertex in nulliparous women in active labor. A risk factor for cesarean delivery. AB - OBJECTIVE: To compare the route of delivery among nulliparous parturients with and without an engaged vertex in the early, active phase of labor. METHODS: Prospectively, the position of the fetal head was ascertained among nulliparous women at 37 weeks' gestation or more in early, active labor (cervical dilation > or = 4 cm with adequate contractions). Sixteen variables, including maternal demographics, obstetric complications and intrapartum course, were examined using chi 2 and logistic regression analysis. RESULTS: Among the 77 patients, 33 (42.8%) had an unengaged vertex and 44 (57.2%) had an engaged vertex in active labor. Of the 22 cesarean deliveries for arrest disorder, 2 were in the engaged and 20 in the unengaged group (P < .001). The mean birth weight was similar among those who had vaginal (3,211 +/- 416 g) and cesarean delivery (3,400 +/- 489 g, P = .08). Univariate analysis indicated that chorioamnionitis (relative risk [RR] 2.6, 95% confidence interval [CI] 1.4-4.9) and unengaged vertex (RR 13.3, CI 3.3 53.0) were associated with cesarean delivery for arrest disorders. When entered into a multiple logistic model, only unengagement was a risk factor for cesarean delivery. The following were not associated with cesarean delivery: maternal demographics, gestational age, estimate of fetal weight, presence or absence of meconium, preeclampsia, diabetes mellitus, private obstetric care or use of epidural anesthesia. CONCLUSION: Among nulliparous parturients, an unengaged vertex is a significant risk factor for cesarean delivery for arrest disorders. PMID- 9749417 TI - Differentiating vulvar intraepithelial neoplasia from nonneoplastic epithelial disorders. The toluidine blue test. AB - OBJECTIVE: To determine the effectiveness of the toluidine blue test in the differentiation of vulvar intraepithelial neoplasia (VIN) and nonneoplastic epithelial disorders (NNEDs). STUDY DESIGN: This retrospective clinical study included all women with VIN (n = 24) and NNED (n = 72) referred to a vulvar clinic at a university hospital during a two-year period. Vulvoscopy, staining of vulvar epithelium with 1% toluidine blue and punch biopsy were performed. RESULTS: Vulvar epithelium demonstrated toluidine blue staining in 100% of the patients with VIN 3, in 83% of women with VIN 1-2, in 50% of the women with squamous cell hyperplasia and in 10% of the women with lichen sclerosus. The differences in staining between the groups were statistically significant (P < .001). The sensitivity of toluidine blue staining for the detection of VIN was 92%; the negative predictive value 96% in teh investigated cohort. The specificity for strong staining was 88%. CONCLUSION: The toluidine blue test is an inexpensive and reliable method of separating VIN from hyperplastic NNED areas and choosing a biopsy site on the vulva. PMID- 9749420 TI - Urine protein dipstick measurements. A screen for a standard, 24-hour urine collection. AB - OBJECTIVE: To determine if the sum of urine protein dipstick values recorded during every void can be used to screen for patients who need a standard, 24-hour urine collection for determination of protein excretion. STUDY DESIGN: Thirty inpatient and 17 outpatient pregnant women undergoing 24-hour urine collection for protein concentration were enrolled. The volume, dipstick protein values and time of void were recorded. The 24-hour quantitative analysis of protein excretion performed by the laboratory was compared to the 24-hour sum of the products of each voided volume and dipstick value (dipstick factor). RESULTS: The dipstick factor significantly correlated with the total 24-hour protein excretion (correlation coefficient 0.84, P < 1 x 10(-7)). A dipstick factor of > or = 300 mg, indicated proteinuria with a sensitivity of 96%, specificity of 90%, positive predictive value of 92% and negative predictive value of 95%. Separately, we found differences in the amount of protein excretion when the 24-hour period was divided into six 4-hour periods, using analysis of variance. Paired t test analysis of the mean protein excretion from 16:00 to 04:00 showed significantly higher results than did the protein excretion from 04:01 to 16:00 (1,197 +/- 356 mg vs. 674 +/- 158 mg, P < .0001). CONCLUSION: The sum of dipstick factors in a 24-hour period is a reliable screening test for identifying patients who need the standard laboratory test for proteinuria. PMID- 9749419 TI - Gamete intrafallopian transfer. Comparison of epidural vs. general anesthesia. AB - OBJECTIVE: To investigate the reproductive outcome of laparoscopic gamete intrafallopian transfer (GIFT) performed under epidural vs. general anesthesia. STUDY DESIGN: Retrospective analysis of 110 consecutive laparoscopic GIFT procedures performed under general or epidural anesthesia. All patients underwent controlled ovarian hyperstimulation employing human menopausal gonadotropin and gonadotropin releasing hormone agonist, given either in the midluteal or early follicular phase. Data were retrieved concerning age, diagnosis, estradiol levels and maximum follicular diameter at the time of human chorionic gonadotropin injection, percentage of mature oocytes retrieved and number of oocytes transferred. These variables were compared using the independent means t test. Pregnancy rates and outcome between the general and epidural anesthesia groups were compared with the chi 2 test. RESULTS: Of the 110 procedures, 84 were performed under general anesthesia, whereas 22 were done using epidural anesthesia. Four procedures started with epidural anesthesia and were converted to general anesthesia because of upper abdominal discomfort. The success rate of epidural anesthesia, therefore, was 85% (22/26). There were no significant differences in the confounding variables between the general and epidural anesthesia groups. However, patients receiving epidural anesthesia had a significantly higher pregnancy rate, 59.1%, and a live birth rate of 40.9% as compared to 31.0% and 21.4%, respectively, for the general anesthesia group. CONCLUSION: Laparoscopic GIFT can be performed safely under epidural anesthesia. Because of the higher pregnancy and live birth rates, epidural is the anesthetic of choice for GIFT. However, a prospective, randomized study is needed to confirm the above observation. PMID- 9749421 TI - Reproductive health in diabetic women. A report of two cases demonstrating the importance of preconception care. AB - BACKGROUND: Optimal glycemic control prior to and during early pregnancy is essential in diabetic women. CASES: Two cases of fetal anencephaly occurred in diabetic women who conceived with medical technical assistance but did not receive preconception counseling. Both women had poor glycemic control periconceptually, as evidenced by elevated glycosylated hemoglobin determinations in the early first trimester. Targeted sonographic evaluations in the second trimester revealed both fetuses to be anencephalic. Both women terminated their pregnancies. CONCLUSION: Women with diabetes should be encouraged to seek preconceptual counseling and achieve tight glycemic control prior to attempting pregnancy. Health care providers play a vital role in stressing the importance of preconception care to these patients. PMID- 9749422 TI - Failure of isotretinoin and interferon-alpha combination therapy for HPV-linked severe vulvar dysplasia. A report of two cases. AB - BACKGROUND: Retinoids (RA) and interferon (IFN) have been reported to be active against a variety of tumors and human papillomavirus (HPV)-related lesions. Because chronic and recurrent HPV-linked vulvar intraepithelial neoplasia 3 (VIN 3) have a high risk of invasion, we evaluated combined therapy of IFN-alpha with 13-cis-retinoic acid (13 cRA) in the treatment of two VIN 3 cases of this type. CASE: Two patients with chronic and recurrent VIN 3 were treated with combined therapy of IFN-alpha (4.5 x 10(6) five times a week) and 13 cRA (1 mg/kg/d) for six months. Clinical regression was observed at the end of treatment in both cases, but histologic features of VIN 3 were still present. CONCLUSION: These data demonstrate the ineffectiveness of the combined regimen of IFN-alpha and 13 cRA with this schedule for a period of six months in recurrent and chronic VIN 3. PMID- 9749423 TI - Hysteroscopic cervical cannulation under ultrasound guidance. A case report. AB - BACKGROUND: Ultrasound guidance has been recommended for various operative hysteroscopic procedures as an alternative to laparoscopic guidance. Ultrasound is noninvasive and may decrease the incidence of uterine perforation. CASE: A 30 year-old nulligravida presented for the evaluation of amenorrhea of two months' duration. She was diagnosed as having cervical obstruction and underwent operative hysteroscopy with cervical cannulation under ultrasound guidance. The patient's menstrual flow returned at the appropriate time without dysmenorrhea. CONCLUSION: Ultrasound guidance during hysteroscopy assisted in the proper orientation and position of the hysteroscope at the time of cannulation, potentially minimizing the risk of uterine perforation. PMID- 9749425 TI - Successful pregnancy in a woman with acute myeloid leukemia treated with high dose whole-body irradiation. AB - BACKGROUND: Although radiotherapy is an integral part of managing certain types of hematologic malignancies, its effect on the reproductive system are well established. We report a case of successful pregnancy in a patient who received high-dose whole-body irradiation (WBI) (1,575 cGy) as part of her treatment for acute myeloid leukemia (AML). CASE: A 26-year-old woman received high-dose cyclophosphamide accompanied by high-dose (1,575 cGy) WBI as part of her treatment for AML when she was 23 years of age. The patient received oral contraceptives before, during and after treatment. After WBI, the patient developed ovarian failure and amenorrhea, which was confirmed by hormonal evaluation. The amenorrhea persisted for one year. No recurrence of AML was found. The patient was placed on hormone replacement therapy (HRT) because of vasomotor changes. An unexpected pregnancy occurred 14 months later; HRT was discontinued. The patient delivered a normal female infant at 38 weeks of gestation. The infant was followed for eight months; her development appeared to be normal. CONCLUSION: In this case report, it is unclear whether pregnancy resulted from active folliculogenesis remote from radiation therapy or from possible ovarian protection rendered by the use of oral contraceptives. The benefit of oral contraceptives in protecting the ovary from radiation injury is unknown and remains an area for future research. PMID- 9749424 TI - Laparoscopically assisted extracorporeal neosalpingostomy. A case report. AB - BACKGROUND: Laparoscopic neosalpingostomies can be time consuming, costly and technically difficult, resulting in suboptimal results in inexperienced hands. Extracorporeal techniques for pelvic and abdominal organs have already been previously reported in the literature, involving such organs as the appendix, ovary and small bowel. CASE: A 33-year-old white woman, gravida 0, diagnosed with a history of primary infertility, underwent diagnostic laparoscopy/hysteroscopy. Stage IV endometriosis was diagnosed and was accompanied with bilateral tubal occlusion with complete obliteration of the fimbriae bilaterally. Extracorporealization of the fallopian tubes was performed followed by microsurgical neosalpingostomies. Operative time for both neosalpingostomies was 20 minutes. CONCLUSION: When controlled for tubal mucosa integrity, previous data from one researcher comparing microsurgical laparotomy and laparoscopic neosalpingostomies shows a higher fertility rate with open surgical techniques. This may be explained, in part, by suboptimal repair of the distal tubes by a laparoscopic technique. This extracorporeal technique may aid in shorter surgery and anesthesia times, higher intrauterine pregnancy rates and higher successful surgical completions. Prospective evaluation of multiple patients will be necessary to assess its efficacy. Even if pregnancy rates do not differ, operative time and cost may be significantly reduced. PMID- 9749426 TI - Bilateral tubal pregnancy. A report of an unusual case. AB - BACKGROUND: Bilateral tubal pregnancies are rare and are usually confirmed simultaneously during the same operation. We report a case in which the right salpingectomy was performed seven weeks before the left salpingectomy. When the right salpingectomy was done, the left uterine tube appeared entirely normal. CASE: A 38-year-old woman underwent laparoscopic surgery for suspected right tubal pregnancy. A right tubal pregnancy was found to have partially aborted into the peritoneal cavity. The left uterine tube was carefully inspected and appeared normal. Histopathology of the right tube showed products of conception and chorionic tissue. Seven weeks after surgery, the woman presented in hemorrhagic shock necessitating emergency laparotomy and left salpingectomy. Histopathology of the left tube confirmed the presence of chronic tissue. The patient did not have coitus between the two salpingectomies. CONCLUSION: The explanation of the presentation is uncertain. However, this case underscores the importance of careful follow-up of patients after laparoscopic surgery for ectopic pregnancies. PMID- 9749427 TI - Successful prenatal digoxin therapy for Ebstein's anomaly with hydrops fetalis. A case report. AB - BACKGROUND: Ebstein anomaly is a rare tricuspid valve anomaly. Some fetuses with Ebstein's anomaly have concurrent severe cardiac function impairment, which results in hydrops fetalis. Most of these fetuses are inevitably terminally ill. No reports have demonstrated the potential prenatal therapy for fetuses under such conditions. CASE: Ebstein's anomaly and hydrops fetalis were detected at 28 weeks' gestation. Tricuspid regurgitation with congestive heart failure was observed. From 28 to 34 weeks' gestation, intrauterine therapy with digoxin, 0.75 mg/d, was administered. The fetal hydrops status improved gradually, while the tricuspid valve regurgitation persisted. At 36 weeks' gestation the fetus was delivered normally. During the neonatal phase, digoxin was continued and gradually tapered off. The tricuspid valve regurgitation and cardiomegaly gradually improved. CONCLUSION: The favorable outcome in this case supports the positive effect of prenatal digoxin therapy for Ebstein's anomaly with hydrops fetalis. In such conditions, upon the appearance of hydrops and congestive cardiac failure, immediate digoxin therapy may be useful. This successful trial encouraged us to manage such fetuses more aggressively. PMID- 9749429 TI - Massive hemoperitoneum from endometriosis of the fallopian tube. A case report. AB - BACKGROUND: Endometriosis is a common gynecologic disease that usually presents with pelvic pain and infertility in the reproductive years. It can be complicated by bleeding, such as hematuria or hemoptysis; however, acute massive hemoperitoneum caused by tubal endometriosis without any concomitant disorder has not been reported previously. CASE: An unusual case of massive hemoperitoneum led to preshock as a result of bleeding from a tubal endometriosis implant in a previously healthy 29-year-old woman without previous history suggesting endometriosis. CONCLUSION: Although the most common gynecologic cause of hemoperitoneum in a reproductive-age woman is ruptured ectopic pregnancy, endometriosis should also be considered, especially after exclusion of pregnancy. PMID- 9749428 TI - Severe pyruvate kinase deficiency anemia. A case report. AB - BACKGROUND: Pyruvate kinase deficiency is a rare cause of hemolytic anemia and, in its most severe form, requires splenectomy in childhood. During pregnancy, severe cases have been traditionally managed with prophylactic blood transfusions to keep the hemoglobin concentration above arbitrary thresholds of 7-8 g/dL. CASE: A case of severe pyruvate kinase deficiency anemia was managed conservatively without blood transfusions even though the hemoglobin concentration reached a nadir of 6.8 g/dL. The perinatal outcome was good. CONCLUSION: In cases of severe pyruvate kinase deficiency anemia, pregnancy per se might not be an indication for prophylactic blood transfusions. PMID- 9749430 TI - Successful treatment of advanced interstitial pregnancy with methotrexate and hysteroscopy. A case report. AB - BACKGROUND: Data concerning medical treatment of interstitial ectopic pregnancies are scarce. These pregnancies are characterized by late and serious clinical manifestations. We report a case of advanced interstitial pregnancy treated successfully by combining methotrexate (MTX) and hysteroscopy. CASE: A routine ultrasonic evaluation of a 10-week pregnancy revealed a right interstitial gestational sac 58 mm in diameter and containing an embryo with a crownrump length of 29 mm and embryonic heartbeats. Serum beta-human chorionic gonadatropin (hCG) level was 97,950 mIU/mL. The patient was treated with a systemic MTX/leucovorin regimen. At the end of the one-week course, no embryonic cardiac activity was detected, and a decrease in beta-hCG levels commenced. Persistent trophoblastic tissue, manifested by a low (26 mIU/mL) beta-hCG level in plateau, was successfully removed by way of hysteroscopy. CONCLUSION: Early detection of interstitial pregnancy may facilitate conservative medical treatment. PMID- 9749431 TI - [Physiologo-biochemical characteristics if Gluconobacter oxydans and prospects for its use in biotechnology and biosensor systems (review)]. AB - Gluconobacter oxydans possesses a unique organization of metabolic systems, which are characterized by reduction of major dissimilation pathways, surface localization of main oxidative enzymes responsible for partial oxidation of carbon substrates, high performance of electron-transport chains, and accumulation of partially oxidized metabolites in the medium. These features allow us to use the cells of these microorganisms in biotechnology for production of several food products and medicines. The use of G. oxydans in biosensors for estimation of concentrations of sugars, aldoses and polyalcohols is promising. Physiological and biochemical features of these microorganisms enabling their use in biotechnology and receptor elements of biosensors are reviewed. PMID- 9749432 TI - [Development of microbiological technology of air deodoration in laboratory industrial conditions using a pilot plant]. AB - Laboratory tests were performed to select a complex of bacterial strains capable of effective deodoration of waste air produced by an animal formulated feed works under elevated temperature with the presence of numerous organic pollutants. The complex included species from the general Nocardia, Rhodococcus, and Comamonas. The biocatalyst was tested in a real industrial process with the use of a pilot plant for microbiological deodoration of waste air. The test lasted for over six months and confirmed the efficiency of the development method of deodoration. PMID- 9749434 TI - [Simplified model of increase in colony diameter during growth of unicellular microorganisms and its use in evaluating the effect of biocides on microbial cells]. AB - A simplified model of increase in colony diameter is proposed. The model uses the size of single cells and several measurements of colony diameter during linear growth for calculating with good approximation the growth curve for the culture from the moment of inoculation. The parameter mu(m)', which is approximately 10% lower than the maximum specific growth rate of the colony biomass, could be also calculated. The effect of copper sulfate on the colony growth of Pseudomonas sp. G-1 was studied using the model. A high concentration of Cu2+ ions was found to result in decreases in the value of mu(m)', colony diameter, and the rate of increase in the colony diameter. PMID- 9749433 TI - [Optimization of hydrolysis of protein substrates by yeast proteases from Saccharomyces carlsbergenis]. AB - Optimum conditions for enzymatic hydrolysis of bovine blood in the presence of brewer's yeast Saccharomyces carlsbergensis were studied. In addition to the amount of blood, yeast, sodium citrate, and ethanol in the mixture, the optimum process duration and temperature were determined. The yield of the major end product (blood hydrolysate) increased from 11 to 55%, and the amino acid composition of the end product was brought to a balanced level as a result of hydrolysis optimization and addition of pancreatin to the reaction mixture. PMID- 9749435 TI - [Coimmobilization on unmodified polymers of the proteolytic enzyme protease S and antimicrobial drugs]. AB - In order to develop materials of prolonged biological effects, the proteolytic enzyme protease S was coimmobilized with antimicrobial drugs chlorhexidine bigluconate and gentamicin sulfate on unmodified cellulose and polyester fibrous materials. The drugs were incorporated into the polymeric composition, which was attached to the materials studied. PMID- 9749436 TI - [Hypochromic effect of signal attenuation in 31P-NMR spectra of linear polyphosphates]. AB - Chemical shifts in 31P-NMR spectra of linear polyphosphates were studied. In each polyphosphate species tested, the sum of signal intensities of the internal (core) phosphate groups was proportional to the concentration of each polyphosphate, but the contribution of such groups to the total intensity of the signal decreased with increasing the length of the polyphosphate chain. An equation for estimating the polyphosphate chain length in biological objects taking into account a decrease in the 31P-NMR spectral intensity is proposed. PMID- 9749437 TI - [Antiprotease and immunostimulating effect of peptide components isolated from cuticle of the chicken stomach]. AB - Peptides with molecular weights of 0.8 to 16 kDa were isolated from the keratinous envelope of chicken muscular stomach cuticle. The peptide preparation had a immunostimulating activity whose qualitative and quantitative traits made it comparable with the activity of known drugs of similar composition. In addition, the peptide preparation reversibly inactivated trypsin and activated chymotrypsin. A low-molecular-weight protein (14.1 kDa) was isolated from the peptide preparation by affinity chromatography on trypsin-Sepharose 4B. This protein was found to be a reversible noncompetitive inhibitor of trypsin. PMID- 9749438 TI - Statistical shortcomings in licensing applications. AB - This paper concerns the statistical work carried out with respect to clinical trials conducted for regulatory purposes. Although the general quality of such work has improved markedly over recent years and is now generally high, a number of shortcomings remain. A few of these arise from failure to follow well established statistical practice. Rather more arise from a poor understanding of areas of known statistical disagreement and from the unsatisfactory use of newer and more advanced techniques. Inadequacies in reporting statistical work are commonplace. Examples of all these shortcomings are provided and emphasis is placed on the value of a statistical contribution to overall summaries such as the clinical expert report. PMID- 9749440 TI - Practical issues in equivalence trials. AB - Equivalence trials aim to show that two treatments have equivalent therapeutic effects. The approach is to define, in advance, a range of equivalence -d to +d for the treatment difference such that any value in the range is clinically unimportant. If the confidence interval for the difference, calculated after the trial, lies entirely within the interval, then equivalence is claimed. Glaxo Wellcome has carried out a series of trials using this methodology to assess new formulations of inhaled beta-agonists and inhaled steroids in asthma. Eleven of these trials are used to review some practical issues in equivalence trials. For the series of asthma trials, a range for peak expiratory flow rate (PEF) from -15 to +15 l/min was chosen to be the range of equivalence. This fitted well with physicians' opinions and with previously demonstrated differences between active and placebo. The choice of the size of the confidence interval should depend on the medical severity of the clinical endpoints under consideration and the level of risk acceptable in assuming equivalence if a difference of potential importance exists. From this point of view, a recommendation in the CPMP Note for Guidance on Biostatistics that 95 per cent confidence intervals should be used is inappropriate. Intent-to-treat (ITT) and per-protocol (PP) analyses were compared for the eleven asthma trials. Confidence intervals were always wider for the PP analysis and this was entirely due to the smaller number of subjects included in the PP analysis. There was no evidence that the ITT analyses were more conservative in their estimates of treatment difference. The need to demonstrate equivalence in both an ITT and a PP analysis in a regulatory trial increases the regulatory burden on drug developers. The relative importance of the two analyses will depend on the definitions used in particular therapeutic areas. Demonstrating equivalence in one population with strong support from the other would be preferred from the Industry viewpoint. In trials with regulatory importance, prior agreement with regulators on the role of ITT and PP populations should be sought. Trial designs will need to take account of the estimated size of the PP population if adequate power is needed for both analyses. Careful design in the series of asthma trials, particularly identifying a population of patients with potential to improve, resulted in notable increases in lung function during the course of the trials for both treatments. This provided reassurance that equivalence was not due to a lack of efficacy for both treatments. In one trial equivalence was demonstrated overall but a treatment by country interaction was noted. However, this interaction could not be attributed to differences in patient characteristics or baseline data between the countries. Study conduct was also similar in the different countries. The conclusion was that the interaction was spurious and that the trial provided good evidence of equivalence. PMID- 9749439 TI - Methodological advances and plans for improving regulatory success for confirmatory studies. AB - Regulatory success for new medical products requires convincing statistical results from planned analyses for relevant and reliable data from well-designed clinical studies. Statistical methodology has a critical role for such success through strengthening the robustness of study findings to sources of bias, variability and spurious events. Areas of particular importance for statistical methodology include the structure for study design, the schedule and procedures for data collection, and plans for primary data analyses. This paper discusses some challenging statistical issues for these areas and suggests potential strategies for addressing them. The objective of these strategies as well as other methodological advances is the minimization of ambiguity in the findings of confirmatory studies. In this way, their effective use can enhance regulatory success. PMID- 9749441 TI - Therapeutic equivalence investigations: statistical considerations. AB - Therapeutic equivalence studies still present problems to regulatory reviewers from many perspectives. This paper is intended to discuss some of these concerns from the statistical viewpoint. There are, however, also some newer approaches which may be particularly useful for the investigation of therapeutic equivalence. PMID- 9749443 TI - A forecasting approach to accelerate drug development. AB - The clinical phase of drug development should be concluded sooner and at a lower cost if primarily only the pivotal and supportive studies were to be conducted. Such improved efficiency requires development of a decision support system that delivers five new capabilities: (i) it enables one to predict a result of a clinical study and to identify those studies that are expected to have an acceptable probability of success; (ii) it will allow one to optimally utilize available pharmacokinetic and pharmacodynamic (PK/PD) data and improve its predictive capability as more data become available; (iii) it will enable one to project useful population results, not just mean results; (iv) predictions will be accompanied by a measure of reliability; and (v) expected initial clinical results will be predictable from animal and related drug class data. With such a tool population targets could be specified very early in the drug development programme, challenged, and then rationally revised at each step during the development process. This report describes progress in developing and testing a clinical trials Forecaster, a prototype for such a system. The Forecaster generates estimates of the joint density for a population of combined PK/PD parameters. That population then serves as a surrogate for the population of individuals. When the resulting joint density is sampled, the obtained sets of parameters may be used to generate data that is statistically indistinguishable from the original experimental data. Such simulated data can be used to validate assumptions, and make inferences on specified population targets that are accompanied by a measure of prediction reliability. We demonstrate use of the forecaster by employing N = 22 PK/PD parameter sets for an orally administered analgesic. PMID- 9749442 TI - Issues for statisticians in pharmaco-economic evaluations. AB - Economic evaluations of new pharmaceutical products are of increasing importance to pharmaceutical companies. In this paper we investigate a number of topics of greater or lesser importance to statisticians who need to involve themselves in pharmaco-economic evaluations. These range from the need to consider whether traditional randomized clinical trials provide the most appropriate setting for an economic evaluation, to the more technical question of how to handle cost effectiveness ratio data, including the issue of the most appropriate inferential apparatus-hypothesis testing, confidence intervals or Bayesian methods. PMID- 9749444 TI - Counts and times to events. AB - A common response recorded in medical statistics involves the occurrence of events. Often this is summarized as a count for each patient. Reasons are given for disaggregating the data as much as possible and instead studying the time to each event. Global counts may hide evolution of the state of the patient over time. This will appear as overdispersion which may wrongly be interpreted as differential frailty among patients. Interval censoring is only important if the rate of events is very high as compared to the time unit of observation chosen. Repeated times to events on the same patient will be interrelated and so should be appropriately treated, as for any repeated measure. Both serial dependence among successive times to events on the same patient and frailty among patients may be present. PMID- 9749445 TI - Some controversies in planning and analysing multi-centre trials. AB - It is shown that a rational approach to planning multi-centre trials will lead to an unequal distribution of patients across centres. Consequently different approaches to estimation will yield different estimates. However, some such approaches are not reasonable and it is concluded that multi-centre trials are less problematic than is commonly supposed. PMID- 9749446 TI - A comparison of various estimators of a treatment difference for a multi-centre clinical trial. AB - When a clinical trial is conducted at more than one centre it is likely that the true treatment effect will not be identical at each centre. In other words there will be some degree of treatment-by-centre interaction. A number of alternative approaches for dealing with this have been suggested in the literature. These include frequentist approaches with a fixed or random effects model for the observed data and Bayesian approaches. In the fixed effects model, there are two common competing estimators of the treatment difference, based on weighted or unweighted estimates from individual centres. Which one of these should be used is the subject of some controversy and we do not intend to take a particular methodological position in this paper. Our intention is to provide some insight into the relative merits of the indicated range of possible estimators of the treatment effect. For the fixed effects model, we also look at the merits of using a preliminary test for interaction assuming a 10 per cent significance level for the test. In order to make comparisons we have simulated a 'typical' trial which compares an active drug with a placebo in the treatment of hypertension, using systolic blood pressure as the primary variable. As well as allowing the treatment effect to vary between centres, we have concentrated on the particular case where one centre is out of line with the others in terms of its true treatment difference. The various estimators that result from the different approaches are compared in terms of mean squared error and power to reject the null hypothesis of no treatment difference. Overall, the approach that uses the fixed effects weighted estimator of overall treatment difference is recommended as one that has much to offer. PMID- 9749448 TI - The management of interim analyses in drug development. AB - The ethical need for interim analyses in long-term trials of life-threatening conditions is well established, but recently there has been increased enthusiasm for the use of interim analyses in other areas of clinical research. Interim analysis is seen as a powerful tool in drug development to help reduce 'time to market' by allowing key decisions to be made earlier on the basis of one or more ongoing clinical trials. However, this goal can only be achieved without serious penalties if interim analyses are properly designed and executed, so that the integrity of the whole drug development programme is maintained. The purpose of this paper is to define procedures for the design and management of clinical trials which involve interim analyses. These procedures are intended to incorporate good clinical and statistical practice, hence maintaining high scientific standards not only of individual trials but also of the overall development programme. Aspects of design, conduct, analysis and dissemination of results are considered, with particular focus on the effective use of data monitoring committees in confirmatory efficacy trials. PMID- 9749450 TI - A framework establishing clear decision criteria for the assessment of drug efficacy. AB - Much has been published on various aspects of data analysis and reporting from clinical trials within the biopharmaceutical environment. This ranges from regulatory guidelines on the format and content of registration dossiers to recommendations on data presentation and the statistical methodologies that are appropriate for the diverse types of data one observes in clinical trials. Little has been written about designing a clinical trial analysis and reporting package that focuses on the decisions that must be made throughout the drug development process. Pharmaceutical companies today are under enormous pressure to develop drugs quickly and (cost-) efficiently. Because of this, drugs often move into the later phases of drug development before evidence from prior phases is completely understood. This provides a challenge to clinical trialists to design and execute a clinical trial programme which can expedite drug development. The statistician, as a clinical trialist, must strive to determine the optimum analytical methodology that facilitates decision making for this clinical trial programme. This paper proposes a new framework for the assessment of efficacy in drug development called the 'one programme, one p-value' framework. This framework will accelerate drug development by providing clear criteria for the decisions which must be made along the way. The 'one programme, one p-value' framework is based on the notion that the clinical trial programme comprises exploratory and confirmatory phases. The use of the likelihood function in the exploratory phase facilitates the decision whether (or when) to move into the confirmatory phase. The confirmatory phase consists of one confirmatory trial with a single hypothesis test of the drug's efficacy; hence 'one p-value'. Sponsor interaction with regulatory agencies is necessary at each decision point. Finally, the paper considers how analysis and reporting of efficacy data can be accomplished from a clinical trial programme as described. PMID- 9749449 TI - Providing evidence of efficacy for a new drug. AB - There are many issues to consider when designing an efficacy package for drug registration. Generally in Europe and the United States, two or more confirmatory trials demonstrating efficacy (p < 0.025, one-tailed) of the test treatment versus a suitable control group must be conducted with a priori definition of a primary efficacy endpoint. Exceptions are possible, and there is always extensive discussion whenever less is proposed or more is required. Every aspect of the basic requirement can be questioned: number of trials; choice of control groups; selection of primary efficacy variables(s); levels of significance; one-tailed versus two-tailed test. These issues will be discussed, and justification is given when proposals are made for deviations from standard practice. Differences between Europe and the U.S. are discussed for certain disease entities. Because the assessment of the weight of evidence in favour of a drug effect is difficult to quantitate, if not impossible, no definitive guidance can be given that is suitable for all circumstances and countries. PMID- 9749447 TI - Multi-centre trial analysis revisited. AB - Analyses of multi-centre trials must consider the effects of the individual centres and the possibility of non-constancy of treatment effect differences among centres. This usually means an ANOVA with terms for centres, treatments, and centre x treatment interactions in practice, at least in the U.S.A. Empirical and conventional Bayes methods provide attractive alternatives to conventional ANOVAs for analysing and reporting the findings from multi-centre trials and do not require more restrictive assumptions than the ANOVA approach. These approaches require regarding the centre effects as random instead of fixed, a view which often will reasonably describe outcomes of clinical trials in spite of the fact that the individual centres certainly do not comprise a random sample of all possible centres. The components of these approaches are well understood and have been employed in related applications such as meta-analysis. Combining them in a way that makes their application to routine multi-centre trial analysis relatively straightforward does not appear to have been described previously, and is what forms the topic of this paper. The empirical Bayes approach leads to useful graphical displays, including one with the data superimposed on probability contours of the joint distribution of the individual centre means and standard deviations, which provides a handy way to identify possible outliers. Covariates can be incorporated without difficulty. The Bayes approach, implemented with Gibbs sampling, provides a convenient way to construct posterior and predictive distributions for a variety of useful statistics. We compare the result of empirical and conventional Bayes analyses with the result of fixed and mixed model ANOVAs applied to data from a multi-centre trial. PMID- 9749451 TI - Statistical issues in pharmacoepidemiological case-control studies. AB - Pharmacoepidemiology is the study of the use and the effect of drugs in large numbers of people. The work relies largely on standard epidemiological principles, which, applied in a new context, bring particular constraints and opportunities. In January 1994, a cluster of children with cystic fibrosis (CF) who developed strictures in the colon was reported. It was suggested that these may have been associated with high strength pancreatic enzyme preparations which had been introduced into use in these patients approximately one year previously. This hypothesis has been evaluated by a case-control study which confirms the association with high-strength pancreatic enzyme usage. The study is described and is used to illustrate statistical issues to provide a framework for future research in statistical methodology in pharmacoepidemiological case-control studies. PMID- 9749452 TI - Biostatistical considerations in pharmacovigilance and pharmacoepidemiology: linking quantitative risk assessment in pre-market licensure application safety data, post-market alert reports and formal epidemiological studies. AB - This paper deals with a conceptual discussion of a variety of statistical concepts, methods and strategies that are relevant to the quantitative assessment of risk derived from safety data collected during the pre- and post-marketing phase of a new drug's life cycle. A call is made for the use of more standard approaches to the analysis of safety data that are statistically and epidemiologically rigorous and for attempts to link the strategies for pre-market safety assessment with strategies for post-market safety evaluation. This link may be facilitated by recognizing the limitations and complementary roles played by pre- and post-market safety data collection schemes and by linking the quantitative analyses utilized for either exploratory or confirmatory purposes of risk assessment in each phase of safety data collection. Examples are provided of studies specifically designed to evaluate risk in a post approval setting and several available guidelines intended to improve the quality of these studies are discussed. PMID- 9749453 TI - Issues for covariance analysis of dichotomous and ordered categorical data from randomized clinical trials and non-parametric strategies for addressing them. AB - Analysis of covariance is an effective method for addressing two considerations for randomized clinical trials. One is reduction of variance for estimates of treatment effects and thereby the production of narrower confidence intervals and more powerful statistical tests. The other is the clarification of the magnitude of treatment effects through adjustment of corresponding estimates for any random imbalances between the treatment groups with respect to the covariables. The statistical basis of covariance analysis can be either non-parametric, with reliance only on the randomization in the study design, or parametric through a statistical model for a postulated sampling process. For non-parametric methods, there are no formal assumptions for how a response variable is related to the covariables, but strong correlation between response and covariables is necessary for variance reduction. Computations for these methods are straightforward through the application of weighted least squares to fit linear models to the differences between treatment groups for the means of the response variable and the covariables jointly with a specification that has null values for the differences that correspond to the covariables. Moreover, such analysis is similarly applicable to dichotomous indicators, ranks or integers for ordered categories, and continuous measurements. Since non-parametric covariance analysis can have many forms, the ones which are planned for a clinical trial need careful specification in its protocol. A limitation of non-parametric analysis is that it does not directly address the magnitude of treatment effects within subgroups based on the covariables or the homogeneity of such effects. For this purpose, a statistical model is needed. When the response criterion is dichotomous or has ordered categories, such a model may have a non-linear nature which determines how covariance adjustment modifies results for treatment effects. Insight concerning such modifications can be gained through their evaluation relative to non-parametric counterparts. Such evaluation usually indicates that alternative ways to compare treatments for a response criterion with adjustment for a set of covariables mutually support the same conclusion about the strength of treatment effects. This robustness is noteworthy since the alternative methods for covariance analysis have substantially different rationales and assumptions. Since findings can differ in important ways across alternative choices for covariables (as opposed to methods for covariance adjustment), the critical consideration for studies with covariance analyses planned as the primary method for comparing treatments is the specification of the covariables in the protocol (or in an amendment or formal plan prior to any unmasking of the study. PMID- 9749455 TI - [Functional reorganizations of the neuronal structures of the cat associative cortex after total sectioning of the corpus callosum]. AB - In given work results of researches functional organization of neural networks at associative cortex level in intact and after total corpus callosum section (Cc sec) of cat's brain. In result of the given researches is established, that after total Cc sec the brain is exposed significant morphological and functional changes. Is shown, that Cc sec is accompanied by infringements in act of reception of food and head orientation of the animal in space. A configuration and characteristics varies conducting ways, opportunity of an exchange the information between is lost by hemispheres, and accordingly, to receive the information from ipsilateral hemifield of vision. The structural and functional characteristics vary of neural structures of both hemispheres: asymmetry of hemispheres in size disappears, orientational and directional characteristics and decreases number of subzones in receptive fields (RF). In RF of this area occur extensive inhibitory zones. Arrangements RF in vision field main borrows contralateral hemifield of vision, in intact a brain large part RF pass in ipsilateral hemifield of vision. Thus in result of these researches is established, that the majority of neural functions of left and right hemispheres at level associative cortex, revealed on intact brain are formed for account transcallosal of connections. In conditions of isolated functioning both hemispheres there are the changes as in ways of the information processing, and in complementary ways of display of space. In result callosotomy of neuron networks in both hemispheres undergo essential changes: the hemispheres lose ability to integrate the set of the parties of subjects, disturbed an opportunity of invariant the descriptions of images, hence the description of the space information occurs on separate attributes of this information. PMID- 9749454 TI - [The cerebral control of the somatosensory and auditory afferent projections to the cerebral cortex in man and animals]. AB - The main purpose of the present paper was consisted in studying of topology (spreading) of the somatosensory and auditory projections in cortex of both brain hemispheres in humans under different functional states conditions: during quiet walking state and during realization of the meditative programme. At the same time for the purpose of verification these steering mechanisms a number of experiments were realized in animals with neurosurgical cutting of brainstem ascending projections, which control the transfer of somatosensory and auditory sensibility. Experimental study was realized with two groups of subjects--8 subjects (age from 25 to 35 years old) and 25 (age from 25 to 40 years old) subjects practicing technique of Transcendental Meditation (TM). In addition to the mentioned above group some groups of animals were used in the experiments. Among them there were used the groups of monkeys (8 macaque rhesus and macaque nemestrina) and cats (10 animals) in conditions of acute experiment, under tiopenthal anesthesia. Two experimental methods were used in the study: electrophysiological for subjects and neurosurgical, additionally for animals. For evaluation of the brain reactivity in subjects registration of the somatosensory, to median nerve stimulation, and auditory, to bilateral application of auditory clicks, evoked potentials (EPs) in the symmetrical cortical structures of the brain was used. Registration of the somatosensory and auditory evoked potentials in animals was realized not only from the cortex, but from the brainstem somatosensory and auditory structures. SSEP in subjects meditators were registered before and during meditation programme. In animals SSEP and AEP registration on the corresponding stimuli realized before and after neurosurgical operation--section of the midbrain tegmentum. Specific alterations of the early (up to 80 ms) and late components SSEP and AEP complexes in forms of topology spreading and diminution of registration areas of these components were obtained during the meditation. Origins of these functional reorganization are discussed from the positions of internal 'feed-back' inhibition. PMID- 9749456 TI - [The nonstability of the EEG: a methodological and experimental analysis]. AB - Theoretical, methodological and methodical aspects on nonstationary EEGs, that is, EEGs whose patterns undergo changes with time are reviewed. The piecewise description of the EEG is confirmed on the base of the own data and results of another investigators. It is analysed the basic approach to statistical evaluation of nonstationary EEGs in the terms of quasi-stationary segments. The special attention is devoted tho the data of segmental organization of the bioelectrical field of the cortex. Taking into consideration new experimental data concerning the time consistency of the segmental descriptions of regional EEGs it is suggested conception of "operational synchrony" as a form of discretial cooperation of cortex process. The theoretical explanations of the phenomenology for piecewise functioning of neuron nets are discussed. PMID- 9749457 TI - [FMRFa and FMRFamide-like peptides (FaRPs) in the pathogenesis of shock]. AB - The cardioactive FMRFa is found in the several animal tissues but the action mechanism and the physiological role of FMRFa and FaRPs is unknown. Authors discussed the hypothesis that FMRFa and FaRPs may act as endogenous sympatoadrenal system modulators; sometimes they can act as adrenaline-like hormones. Their physiological significance may be more clear by the pathological and hypobiotics conditions. PMID- 9749458 TI - [The functional activity, energetics and survival of the heart under hypothermia]. AB - Adequate blood supply to tissue, which is the most important prerequisite for survival of the organisms when put under hypothermal conditions and taken out of these conditions depends, above all, on the cardial function. Detailed exploration of the automatic reactions, response to stimulation, conductivity and systole of hypothermal cardium using a cardio-pulmonary preparation has been reported by V. M. Pokrovskii and co-authors. As far as we are informed, this monographic volume is the only available review of developments in the last decades in studies of cardium under hypothermal conditions. In this article we shall touch upon the issues not covered by the above monograph including energy status of hypothermal cardium and the issues important the physiological and medical perspectives: cardium functional status in situ under accidental hypothermal conditions (hypothermal freezing), and maintaining functionality of cardium after its stoppage in hypothermal conditions. PMID- 9749459 TI - [The functional specificity of the cardiac pacemaker system]. AB - On the basis of the own and literature data the original conception on pacemaker system of the heart experimentally is grounded. The new classification of the specialized structure--functional formations in the heart is suggested. The functional specificity of pacemaker system is considered in the comparative ontogenetic aspect. PMID- 9749460 TI - [The role of the forestomachs in the adaptation of the alimentary function in ruminants]. AB - Dietary character of ruminants in the north is principally different between "nonselective" (cattle, sheep) and "selective" (reindeer, moose) species. Due to the developed polyfunctioning, forestomach in ruminants, primarily rumen and reticulum, are involved in the homeostasis formation in enteral and interior mediums, nutrients deposition and recirculation. That provides efficiency in utilization of deficient in nutrients forage and body reserves of wild ruminants during the winter. The original functional organization of absorption processes and nutrients recirculation across the multilayer epithelium of forestomach also promotes realization of the two levels metabolism during the year and adaptation of wild ruminants to season dynamics of nutrition. PMID- 9749461 TI - [The artificial joint in the biological environment]. AB - The mechanisms of the joint endoprothesis components evolution and wear as well as development periarthric tissue membrane are discussed. The loosing process is connected with immuno-inflammatory reactions in periarthric tissues on products' wear and modified proteins which activate the bone resorption and/or proliferative reactions as ectopic bone formation. Systemic effects of artificial joints can be connected with migration of metal ions and primated in periarthric tissues of immunocompetent cells. It is concluded that in developing new joint endoprotheses there is a necessity in a systemic approach taking into account all well-known peculiarities of this complex biotribological object at present. PMID- 9749463 TI - A piece of my mind. An independent scientist. PMID- 9749464 TI - International group seeks to dispel incontinence "taboo". PMID- 9749465 TI - New medications aid cognition in schizophrenia. PMID- 9749462 TI - [The factors of individual sensitivity to ultraviolet radiation]. AB - Correlations between effective UVR doses for induction of skin erythema, premature aging and cancer, immunosuppression, vitamin D photobiosynthesis and cataract on the one hand and various ethnic signs, sex, age and metabolic states of organism on the other were considered. Individual UV sensitivity by all criteria mentioned with the exception of immunosuppression and cataract is conditioned by specificity of melanin and DNA metabolism, and risk of skin cancer incidence besides that depends of immune system state. Values of risk factors for UVR effects mentioned incidence in individuals of different sex and age with various ethnic and metabolic conditions are presented. PMID- 9749467 TI - Science groups urge creation of food safety chief. PMID- 9749466 TI - "What are you going to do with a 41-year-old man?". PMID- 9749469 TI - General psychiatry. PMID- 9749468 TI - From the Centers for Disease Control and Prevention. Adoption of hospital policies for prevention of perinatal group B streptococcal disease--United States, 1997. PMID- 9749470 TI - Cigarette smoke exposure and hearing loss. PMID- 9749471 TI - Cigarette smoke exposure and hearing loss. PMID- 9749472 TI - Evaluating antismoking advertising campaigns. PMID- 9749475 TI - The Death of Innocents. PMID- 9749474 TI - The Death of Innocents. PMID- 9749473 TI - Haitian diethylene glycol disaster and Dante's darkwood. PMID- 9749476 TI - Montelukast for children with asthma. PMID- 9749477 TI - Physician marketing of nutritional supplements. PMID- 9749478 TI - Biochemical outcome after radical prostatectomy, external beam radiation therapy, or interstitial radiation therapy for clinically localized prostate cancer. AB - CONTEXT: Interstitial radiation (implant) therapy is used to treat clinically localized adenocarcinoma of the prostate, but how it compares with other treatments is not known. OBJECTIVE: To estimate control of prostate-specific antigen (PSA) after radical prostatectomy (RP), external beam radiation (RT), or implant with or without neoadjuvant androgen deprivation therapy in patients with clinically localized prostate cancer. DESIGN: Retrospective cohort study of outcome data compared using Cox regression multivariable analyses. SETTING AND PATIENTS: A total of 1872 men treated between January 1989 and October 1997 with an RP (n = 888) or implant with or without neoadjuvant androgen deprivation therapy (n = 218) at the Hospital of the University of Pennsylvania, Philadelphia, or RT (n = 766) at the Joint Center for Radiation Therapy, Boston, Mass, were enrolled. MAIN OUTCOME MEASURE: Actuarial freedom from PSA failure (defined as PSA outcome). RESULTS: The relative risk (RR) of PSA failure in low risk patients (stage T1c, T2a and PSA level < or =10 ng/mL and Gleason score < or =6) treated using RT, implant plus androgen deprivation therapy, or implant therapy was 1.1 (95% confidence interval [CI], 0.5-2.7), 0.5 (95% CI, 0.1-1.9), and 1.1 (95% CI, 0.3-3.6), respectively, compared with those patients treated with RP. The RRs of PSA failure in the intermediate-risk patients (stage T2b or Gleason score of 7 or PSA level >10 and < or =20 ng/mL) and high-risk patients (stage T2c or PSA level >20 ng/mL or Gleason score > or =8) treated with implant compared with RP were 3.1 (95% CI, 1.5-6.1) and 3.0 (95% CI, 1.8-5.0), respectively. The addition of androgen deprivation to implant therapy did not improve PSA outcome in high-risk patients but resulted in a PSA outcome that was not statistically different compared with the results obtained using RP or RT in intermediate-risk patients. These results were unchanged when patients were stratified using the traditional rankings of biopsy Gleason scores of 2 through 4 vs 5 through 6 vs 7 vs 8 through 10. CONCLUSIONS: Low-risk patients had estimates of 5-year PSA outcome after treatment with RP, RT, or implant with or without neoadjuvant androgen deprivation that were not statistically different, whereas intermediate- and high-risk patients treated with RP or RT did better then those treated by implant. Prospective randomized trials are needed to verify these findings. PMID- 9749479 TI - Competing risk analysis of men aged 55 to 74 years at diagnosis managed conservatively for clinically localized prostate cancer. AB - CONTEXT: The appropriate therapy for men with localized prostate cancer is uncertain. Until results of clinical trials are available, men and their physicians need guidance. OBJECTIVE: To estimate survival based on a competing risk analysis stratified by age at diagnosis and histologic findings for men diagnosed as having clinically localized prostate cancer and who were managed conservatively. DESIGN: Retrospective cohort study. SETTING: Connecticut Tumor Registry. PATIENTS: A total of 767 men with localized prostate cancer diagnosed between 1971 and 1984, aged 55 to 74 years at diagnosis, either not treated or treated with immediate or delayed hormonal therapy, and followed up for 10 to 20 years after diagnosis. MAIN OUTCOME MEASURES: Estimates of the probability of dying from prostate cancer or other competing hazards. RESULTS: Men with tumors that have Gleason scores of 2 to 4, 5, 6, 7, and 8 to 10 face a 4% to 7%, 6% to 11%, 18% to 30%, 42% to 70%, and 60% to 87% chance, respectively, of dying from prostate cancer within 15 years of diagnosis depending on their age at diagnosis. CONCLUSIONS: Men whose prostate biopsy specimens show Gleason score 2 to 4 disease face a minimal risk of death from prostate cancer within 15 years of diagnosis. Conversely, men whose biopsy specimens show Gleason score 7 to 10 disease face a high risk of death from prostate cancer when treated conservatively, even when cancer is diagnosed as late as age 74 years. Men with Gleason score 5 or 6 tumors face a modest risk of death from prostate cancer that increases slowly over at least 15 years of follow-up. PMID- 9749480 TI - Chronic multisymptom illness affecting Air Force veterans of the Gulf War. AB - CONTEXT: Gulf War (GW) veterans report nonspecific symptoms significantly more often than their nondeployed peers. However, no specific disorder has been identified, and the etiologic basis and clinical significance of their symptoms remain unclear. OBJECTIVES: To organize symptoms reported by US Air Force GW veterans into a case definition, to characterize clinical features, and to evaluate risk factors. DESIGN: Cross-sectional population survey of individual characteristics and symptoms and clinical evaluation (including a structured interview, the Medical Outcomes Study Short Form 36, psychiatric screening, physical examination, clinical laboratory tests, and serologic assays for antibodies against viruses, rickettsia, parasites, and bacteria) conducted in 1995. PARTICIPANTS AND SETTING: The cross-sectional questionnaire survey included 3723 currently active volunteers, irrespective of health status or GW participation, from 4 air force populations. The cross-sectional clinical evaluation included 158 GW veterans from one unit, irrespective of health status. MAIN OUTCOME MEASURES: Symptom-based case definition; case prevalence rate for GW veterans and nondeployed personnel; clinical and laboratory findings among veterans who met the case definition. RESULTS: We defined a case as having 1 or more chronic symptoms from at least 2 of 3 categories (fatigue, mood-cognition, and musculoskeletal). The prevalence of mild-to-moderate and severe cases was 39% and 6%, respectively, among 1155 GW veterans compared with 14% and 0.7% among 2520 nondeployed personnel. Illness was not associated with time or place of deployment or with duties during the war. Fifty-nine clinically evaluated GW veterans (37%) were noncases, 86 (54%) mild-to-moderate cases, and 13 (8%) severe cases. Although no physical examination, laboratory, or serologic findings identified cases, veterans who met the case definition had significantly diminished functioning and well-being. CONCLUSIONS: Among currently active members of 4 Air Force populations, a chronic multisymptom condition was significantly associated with deployment to the GW. The condition was not associated with specific GW exposures and also affected nondeployed personnel. PMID- 9749481 TI - Association between licensing examination scores and resource use and quality of care in primary care practice. AB - CONTEXT: Clinical competence is a determinant of the quality of care delivered, and may be associated with use of health care resources by primary care physicians. Clinical competence is assumed to be assessed by licensing examinations, yet there is a paucity of information on whether scores achieved predict subsequent practice. OBJECTIVE: To determine if licensing examination scores were associated with selected aspects of quality of care and resource use in initial primary care practice. DESIGN: Prospective cohort study of recently licensed family physicians, followed up for the first 18 months of practice. SETTING: The Quebec health care system. PARTICIPANTS: A total of 614 family physicians who passed the licensing examination between 1991 and 1993 and entered fee-for-service practice in Quebec. MAIN OUTCOME MEASURES: All patients seen by physicians were identified by the universal health insurance board and all health services provided to these patients were retrieved for the 18 months prior to (baseline) and after (follow-up) the physicians' entry into practice. Medical service and prescription claims files were used to measure rates of resource use (specialty consultation, symptom-relief prescribing compared with disease specific prescribing) and quality of care (inappropriate prescribing, mammography screening). Baseline data were used to adjust for differences in practice population. RESULTS: Study physicians saw a total of 1116389 patients, of whom 113535 (10.2%) were elderly and 83391 (7.5%) were women aged 50 to 69 years. Physicians with higher licensing examination scores referred more of their patients for consultation (3.8/1000 patients per SD increase in score; 95% confidence interval [CI], 1.2-7.0; P = .005), prescribed to elderly patients fewer inappropriate medications (-2.7/1000 patients per SD increase in score; 95% CI, -4.8 to -0.7; P=.009) and more disease-specific medications relative to symptom-relief medications (3.9/1000 patients per SD increase in score; 95% CI, 0.3 to 7.4; P= .03), and referred more women aged 50 to 69 years (6.6/1000 patients per SD increase in score; 95% CI, 1.2-11.9; P = .02) for mammography screening. If patients of physicians with the lowest scores had experienced the same rates of consultation, prescribing, and screening as patients of physicians with the highest scores, an additional 3027 patients would have been referred, 179 fewer elderly patients would have been prescribed symptom-relief medication, 912 more elderly patients would have been prescribed disease-specific medication, 189 fewer patients would have received inappropriate medication, and 121 more women would have received mammography screening. CONCLUSIONS: Licensing examination scores are significant predictors of consultation, prescribing, and mammography screening rates in initial primary care practice. PMID- 9749482 TI - Risk factors for pilot fatalities in general aviation airplane crash landings. AB - CONTEXT: Most pilots survive airplane crash landings in small airplanes. Factors associated with pilot death have not been well studied. OBJECTIVE: To identify factors associated with fatalities in general aviation airplane crash landings. DESIGN: Case-control study. SETTING: The United States. SUBJECTS: All pilots in general aviation crash landings of airplanes with 10 seats or fewer, from 1983 through 1992. MAIN OUTCOME MEASURE: Pilot death. RESULTS: Pilots died in 437 (5.2%) of 8411 crash landings. A fire or explosion on the ground was strongly associated with pilot death (relative risk [RR], 20.4; 95% confidence interval [CI], 15.5-26.9), adjusted for pilot age, pilot flight hours, type of landing gear, and the filing of an instrument flight plan. Pilots who failed to use both lap belt and shoulder harness were more likely to die (adjusted RR, 6.8; 95% CI, 1.8-25.5), as were those who used only the lap belt (adjusted RR, 1.7; 95% CI, 1.3-2.2), compared with pilots who used both restraints. CONCLUSION: Pilots may be able to reduce their risk of death in a crash landing by using lap and shoulder restraints. PMID- 9749483 TI - The urgent need to improve health care quality. Institute of Medicine National Roundtable on Health Care Quality. AB - OBJECTIVE: To identify issues related to the quality of health care in the United States, including its measurement, assessment, and improvement, requiring action by health care professionals or other constituencies in the public or private sectors. PARTICIPANTS: The National Roundtable on Health Care Quality, convened by the Institute of Medicine, a component of the National Academy of Sciences, comprised 20 representatives of the private and public sectors, practicing medicine and nursing, representing academia, business, consumer advocacy, and the health media, and including the heads of federal health programs. The roundtable met 6 times between February 1996 and January 1998. It explored ongoing, rapid changes in health care and the implications of these changes for the quality of health and health care in the United States. EVIDENCE: Roundtable members held discussions with a wide variety of experts, convened conferences, commissioned papers, and drew on their individual professional experience. CONSENSUS PROCESS: At the end of its deliberations, roundtable members reached consensus on the conclusions described in this article by a series of discussions at committee meetings and reviews of successive draft documents, the first of which was created by the listed authors and the Institute of Medicine project director. The drafts were revised following these discussions, and the final document was approved according to the formal report review procedures of the National Research Council of the National Academy of Sciences. CONCLUSIONS: The quality of health care can be precisely defined and measured with a degree of scientific accuracy comparable with that of most measures used in clinical medicine. Serious and widespread quality problems exist throughout American medicine. These problems, which may be classified as underuse, overuse, or misuse, occur in small and large communities alike, in all parts of the country, and with approximately equal frequency in managed care and fee-for-service systems of care. Very large numbers of Americans are harmed as a direct result. Quality of care is the problem, not managed care. Current efforts to improve will not succeed unless we undertake a major, systematic effort to overhaul how we deliver health care services, educate and train clinicians, and assess and improve quality. PMID- 9749484 TI - Quality-of-care research: internal elegance and external relevance. PMID- 9749485 TI - Comparing treatments for localized prostate cancer--persisting uncertainty. PMID- 9749486 TI - Illness among Gulf War veterans: risk factors, realities, and future research. PMID- 9749487 TI - Tools for change: CME on the Internet. PMID- 9749489 TI - JAMA patient page: prostate cancer. PMID- 9749488 TI - Teaching hospital costs: implications for academic missions in a competitive market. AB - CONTEXT: As the managed care environment demands lower prices and a greater focus on primary care, the high cost of teaching hospitals may adversely affect their ability to carry out academic missions. OBJECTIVE: To develop a national estimate of total inpatient hospital costs related to graduate medical education (GME). DESIGN: Using Medicare cost report data for fiscal year 1993, we developed a series of regression models to analyze the relationship between inpatient hospital costs per case and explanatory variables, such as case mix, wage levels, local market characteristics, and teaching intensity (the ratio of interns and residents to beds). SETTING AND PARTICIPANTS: A total of 4764 nonfederal, general acute care hospitals, including 1014 teaching hospitals. MAJOR OUTCOME MEASURES: Actual direct GME hospital costs and estimated indirect GME-related hospital costs based on the statistical relationship between teaching intensity and inpatient costs per case. RESULTS: In 1993, academic medical center (AMC) costs per case were 82.9% higher than those for urban nonteaching hospitals (actual cost per case, $9901 vs $5412, respectively). Non-AMC teaching hospital costs per case were 22.5% higher than those for nonteaching hospitals (actual cost per differences in case, $6630 vs $5412, respectively). After adjustment for case mix, wage levels, and direct GME costs, AMCs were 44% more expensive and other teaching hospitals were 14% more costly than nonteaching hospitals. The majority of this difference is explained by teaching intensity. Total estimated US direct and indirect GME-related costs were between $18.1 billion and $22.8 billion in 1997. These estimates include some indirect costs, not directly educational in nature, related to clinical research activities and specialized service capacity. CONCLUSIONS: The cost of teaching hospitals relative to their nonteaching counterparts justifies concern about the potential financial impact of competitive markets on academic missions. The 1997 GME-related cost estimates provide a starting point as public funding mechanisms for academic missions are debated. The efficiency of residency programs, their consistency with national health workforce needs, financial benefits provided to teaching hospitals, and ability of AMCs to maintain higher payment rates are also important considerations in determining future levels of public financial support. PMID- 9749490 TI - Mission impossible? Respond to the challenge. PMID- 9749491 TI - Low rectal cancer: impact of radiation and chemotherapy on surgical treatment. AB - INTRODUCTION: The aim of this study was to evaluate the impact of combined radiotherapy and chemotherapy (leucovorin and 5-fluorouracil) on the treatment of potentially resectable low rectal cancer using the following end points: 1) toxicity of this combined modality regimen; 2) clinical and pathologic response rate and local control; 3) down-staging of the tumor and its influence on the number of sphincter-saving operations; 4) disease-free interval, patterns of relapse, and overall survival. METHODS: From 1991 to 1996, 118 patients with potentially resectable cases of histologically proven adenocarcinoma and no distant metastases were enrolled into this protocol. All patients were evaluated by clinical and proctologic examination, abdominal computed tomography, transrectal ultrasound, and chest radiography. Therapy consisted of 5,040 cGy (6 weeks) and concurrent leucovorin (20/mg/m2/day) with bolus doses of 5 fluorouracil administered intravenously at 425 mg/m2/day for three consecutive days on the first and last three days of radiation therapy. After two months, all patients underwent repeat evaluation and biopsy of any suspected residual lesions or scar tissue. RESULTS: Median follow-up was 36 months. Toxicity of chemotherapy regimen was minimum. Thirty-six patients (30.5 percent) were classified as being complete responders. In six of these patients, complete response was confirmed by the absence of tumor in the surgical specimens (3 abdominoperineal resections and 3 proctosigmoidectomies with coloanal anastomosis). In the remaining 30 patients, confirmation of a complete response was made by the absence of symptoms, negative findings on physical examination, and biopsy, transrectal ultrasound, and pelvic computed tomographic test results during follow-up. Eighty-two patients (69.4 percent) were considered incomplete responders. Residual lesions had already been identified during the first examination in 74 patients. In the other eight patients, residual tumor was only identified after 3 to 14 months. All patients underwent surgical treatment, except one patient who refused surgery. Eighty seven patients underwent 90 surgical procedures: local excision, 9; coloanal anastomosis, 36; abdominoperineal resection, 4; Hartmann's procedure, 1. Isolated local recurrences occurred in five patients (4.3 percent) and combined local and distant failure in eight patients (6.7 percent). Ninety patients are alive and disease-free at a median follow-up of 36 months. CONCLUSIONS: Combined up-front chemoradiotherapy was associated with tolerable and acceptable side effects. A significant number of patients had complete disappearance of their tumors (30.5 percent) within a median follow-up of 36 months. This regimen spared 26.2 percent of patients from surgical treatment and allowed sphincter-saving management in 38.1 percent of patients who may have required abdominoperineal resection. Preliminary results of this trial suggests a reduction in the number of local recurrences and reinforces the concept that infiltrative low rectal cancer may be initially treated by chemoradiotherapy. PMID- 9749492 TI - Conditional survival estimates in 34,963 patients with invasive carcinoma of the colon. AB - PURPOSE: We report colon cancer survival rates that are conditioned on patients having already survived one or more years after diagnosis. These rates have more meaning clinically, because they consider those patients who have already survived a given period of time after treatment. METHODS: The life table method was used to compute conditional survival rates, using population-based data obtained from the Surveillance, Epidemiology, and End Results Program of the National Cancer Institute. Patients were diagnosed between 1983 and 1987 and followed up through 1994. Relative and observed survival rates are considered. RESULTS: Survival rates up to ten years after diagnosis are reported by stage of disease, gender, and race for colon cancer patients who survived from one to five years after diagnosis. Survival rates are also reported by lymph node involvement. CONCLUSIONS: Five-year and ten-year survival in colon cancer patients having already survived between one and five years after diagnosis continues to be influenced significantly by stage and race. PMID- 9749493 TI - Excision of trocar sites reduces tumor implantation in an animal model. AB - PURPOSE: The purpose of this study was to determine the effect of excising abdominal trocar wound sites after pneumoperitoneum on the rate of trocar site tumor implantation in a hamster model. This would help determine whether tumor cells seed trocar sites during or after pneumoperitoneum. METHODS: A total of 0.5 ml of GW-39 human colon cancer cell suspension at 2.5 percent v/v (8 x 10(5) cells) was injected into the abdomens of 77 hamsters through a midline incision. Animals were subjected to ten minutes of pneumoperitoneum, after placement of four abdominal trocars, and then randomly assigned to undergo either simple suture closure or 4-mm radius trocar wound site excision at the end of the procedure. Gross and microscopic tumor implants were documented seven weeks later. RESULTS: There were three and four deaths in simple suture closure and wound site excision groups, respectively. Of the remaining 35 hamsters in each group, tumor cells implanted at 89 and 78 percent of trocar sites, respectively (P < 0.03). There was no significant difference between the two groups in tumor implantation at midline laparotomy sites. Wound site excision also resulted in fewer palpable tumors (44 vs. 61 percent; P < 0.01) and a lower tumor implantation rate (49 vs. 74 percent; P < 0.05) at all four concurrent sites compared with simple suture closure. CONCLUSIONS: Excision of laparoscopic abdominal trocar wound sites can significantly, but not completely, reduce tumor implantation rate compared with simple wound closure. PMID- 9749495 TI - Follow-up of colorectal cancer: a meta-analysis. AB - PURPOSE: The value of intensive follow-up for patients after resection of colorectal cancer remains controversial. This study reviews all randomized and prospective cohort studies to assess the value of aggressive follow-up. METHODS: The literature was searched from the years 1972 to 1996 for studies reporting on the follow-up of patients with colorectal cancer. Randomized and comparative cohort studies that included history, physical examination, and carcinoembrionic antigen values at least three times a year for at least two years were included in a meta-analysis. Single-cohort studies with intensive follow-up and traditional follow-up were also included in a two-group comparative analysis for each outcome indicator. Outcome indicators were 1) curative resection rates after recurrent cancer, 2) survival rates of curative re-resections, 3) length of survival after recurrence, and 4) cumulative five-year survival. RESULTS: Two randomized and three comparative-cohort studies met these criteria and included 2,005 patients, which were evaluated in the meta-analysis. The cumulative five year survival was 1.16 times higher in the intensively followed group (P = 0.003). Two and one-half times more curative re-resections were performed for recurrent cancer in those patients undergoing intensive follow-up (P = 0.0001). Those patients in the intensive follow-up group with a recurrence had a 3.62 times higher survival rate than the control (P = 0.0004). Fourteen single-cohort studies were also included in the comparative analysis of 6,641 patients. The findings from these aggregated studies support the results of the meta-analysis. CONCLUSION: Our study concludes that intensive follow-up detects more recurrent cancers at a stage amenable to curative resection, resulting in an improvement in survival of recurrences and an increased overall five-year cumulative rate of survival. PMID- 9749494 TI - In vitro detection of occult bone marrow metastases in patients with colorectal cancer hepatic metastases. AB - PURPOSE: The purpose of this study was to assess the immunocytochemical status of bone marrow aspirates from patients with clinically isolated hepatic metastases to test the hypothesis that such findings would allow improved patient selection for liver-directed treatment. METHODS: All patients had biopsy-proven or presumed colorectal cancer metastatic to the liver and were scheduled for an operative procedure for hepatic resection or for hepatic artery catheter and chemotherapy pump implant. Immunocytochemical analysis of bone marrow aspirate smears was performed with a panel of monoclonal antibodies directed toward cytokeratins, Lewis Y antigen and A-33 colorectal epitopes. RESULTS: Data from 80 patients indicated that bone marrow reactivity was present in 9.5 percent of those with resectable hepatic metastases and in 34 percent of those not resected (P = 0.03). No single monoclonal antibody or combination produced better discrimination. CONCLUSIONS: Presence or absence of presumed occult colorectal cancer cells in the bone marrow of patients with isolated hepatic metastases is biologically interesting, but not useful in selecting or altering patient management. PMID- 9749496 TI - Role of follow-up in management of local recurrences of colorectal cancer: a prospective, randomized study. AB - PURPOSE: This prospective, randomized, single-center study was designed to evaluate the influence of follow-up on detection and resectability of local recurrences and on survival after radical surgery for colorectal cancer. METHODS: Between 1987 and 1990, 207 consecutive patients who underwent curative resections for primary untreated large-bowel carcinoma were randomly assigned to a conventional follow-up group (Group A; n = 103) and to an intense follow-up group (Group B; n = 104). All the patients were followed up prospectively, and the outcome was known for all of them at five years. Patients in Group A were seen at six-month intervals for one year, and once a year thereafter. Patients in Group B were checked every three months during the first two years, at six-month intervals for the next three years, and once a year thereafter. RESULTS: Of the 103 patients in Group A, local recurrence was detected in 20; 9 (13 percent) of these patients had colon cancer, and 11 (29 percent) had rectal cancer. Of the 104 patients in Group B, local recurrence was detected in 26; 12 (16 percent) of these patients had colon cancer, and 14 (45 percent) had rectal cancer. Twelve cases (60 percent) of local recurrence in Group A and 24 cases (92 percent) in Group B were detected at scheduled visits (P < 0.05). Local recurrences were detected earlier in patients of Group B (10.3 +/- 2.7 vs. 20.2 +/- 6.1 months; P < 0.0003). Curative re-resection was possible in 2 patients (10 percent) in Group A, 1 with colon cancer and 1 with rectal cancer, and in 17 patients (65 percent) in Group B, 6 with colon cancer and 11 with rectal cancer (P < 0.01). Of the Group B patients who had curative re-resections of local recurrence, 8 (47 percent) were disease-free and long-term survivors as of the last follow-up, and 2 (11.7 percent) were alive, but with a new recurrence. The 2 patients in Group A who had curative re-resections died as a result of cancer. The five-year survival rate in Group A was 58.3 percent and in Group B was 73.1 percent. The difference is statistically significant (P < 0.02). CONCLUSIONS: Our data support use of an intense follow-up plan after primary resection of large-bowel cancer, at least in patients with rectal cancer. PMID- 9749497 TI - Intraperitoneal exfoliated cancer cells in patients with colorectal cancer. AB - PURPOSE: The aims of this study were to evaluate potential predictors of exfoliated free cancer cells in the peritoneal cavity and to assess intraoperative peritoneal lavage cytology as a prognostic indicator in patients with colorectal cancer. METHODS: From 1985 to 1987, intraoperative peritoneal lavage cytology was performed in 140 patients with colorectal cancer. Among them, 88 patients underwent curative resection and 52 patients had noncurative surgery. Cytology was examined twice, i.e., immediately after opening the peritoneal cavity (precytology) and just before closing the abdomen (postcytology). One hundred milliliters of saline was poured into the peritoneal cavity and it was retrieved by suction after irrigation. Cytologic examination was performed after staining with Papanicolaou, Giemsa, periodic acid-Schiff, and Alcian blue stains. RESULTS: Among the 140 patients examined, the incidence of positive cytology in the prelavage was 15 percent, and that in the postlavage was 9 percent, although it was 16 percent in either lavage. Among patients with curative resection, 10 percent had positive cytology. Seven characteristics were identified as features of tumors which are prone to exfoliate cells into the peritoneal cavity: 1) macroscopic peritoneal dissemination, 2) liver metastasis, 3) more than 20 ml of ascites, 4) ulcerated tumors without definite borders, 5) invasion of the serosal surface or beyond, 6) semiannular or annular shape, and 7) moderate or marked lymphatic invasion. In patients undergoing curative surgery, among these features, circumferential involvement was the only one correlated closely with positive cytology (P < 0.02). Positive cytology was associated with a worse outcome. In patients who were resected curatively, the postcytology had a stronger influence on local recurrence than the precytology; the local recurrence rate in patients with positive postcytology was higher than in those with negative postcytology, regardless of the precytology. All patients with cancer cells in the peritoneal cavity at the end of surgery had recurrence. CONCLUSIONS: Seven characteristics were identified as risk factors for exfoliation of cancer cells into the peritoneal cavity in patients with colorectal cancer. These findings may be helpful for the choice of laparoscopic surgery in this era of increasing port-site metastases after laparoscopic procedure. The results of peritoneal lavage cytology at the end of surgery were correlated with the long term postoperative outcome of colorectal cancer. Thus, meticulous follow-up and possibly adjuvant chemotherapy may be beneficial for patients with free cancer cells in lavage fluid, even after curative surgery. PMID- 9749498 TI - Anal sphincter integrity and function influences outcome in rectovaginal fistula repair. AB - PURPOSE: Traumatic sphincter disruption frequently is associated with a rectovaginal fistula, but the effect of a persistent sphincter defect on the outcome of rectovaginal fistula repair is poorly documented. We analyzed the outcome of rectovaginal fistula repairs based on preoperative sphincter status. PATIENTS AND METHODS: We identified 52 women who underwent 62 repairs of simple obstetrical rectovaginal fistulas between 1992 and 1995. Fourteen patients (27 percent) had preoperative endoanal ultrasound studies and 25 (48 percent) had anal manometry studies. Follow-up was by mailed questionnaire in 36 patients (69 percent) and by telephone interview in 12 (23 percent), for a total response rate of 92 percent. Median age was 30.5 (range, 18-70) years, and median follow-up was 15 (range, 0.5-123) months. Twenty-five patients (48 percent) complained of varying degrees of fecal incontinence before surgery. There were 27 endorectal advancement flaps and 35 sphincteroplasties (28 with and 8 without levatoroplasty). RESULTS: Success rates were 41 percent with endorectal advancement flaps and 80 percent with sphincteroplasties (96 percent success with and 33 percent without levatoroplasty; P = 0.0001). Endorectal advancement flap was successful in 50 percent of patients with normal sphincter function but in only 33 percent of patients with abnormal sphincter function (P = not significant). For sphincteroplasties, success rates were 73 vs. 84 percent for normal and abnormal sphincter function, respectively (P = not significant). Results were better after sphincteroplasties vs. endorectal advancement flaps in patients with sphincter defects identified by endoanal ultrasound (88 vs. 33 percent; P = not significant) and by manometry (86 vs. 33 percent; P = not significant). Poor results correlated with prior surgery in patients undergoing endorectal advancement flaps (45 percent vs. 25 percent; P = not significant) but not sphincteroplasties (80 vs. 75 percent; P = not significant). CONCLUSIONS: All patients with rectovaginal fistula should undergo preoperative evaluation for occult sphincter defects by endoanal ultrasound or anal manometry or both procedures. Local tissues are inadequate for endorectal advancement flap repairs in patients with sphincter defects and a history of previous repairs. Patients with clinical or anatomic sphincter defects should be treated by sphincteroplasty with levatoroplasty. PMID- 9749499 TI - Hydrogen peroxide-enhanced transanal ultrasound in the assessment of fistula-in ano. AB - Appropriate classification of the fistulous tracts in patients with fistula-in ano may be of value for the planning of proper surgery. Conventional transanal ultrasound has limited value in the visualization of fistulous tracts and their internal openings. Hydrogen peroxide can be used as a contrast medium for ultrasound to improve visualization of fistulas. PURPOSE: This prospective study evaluates hydrogen peroxide-enhanced ultrasound in comparison with physical examination, standard ultrasound, and surgery in the assessment of fistula-in ano. METHODS: Twenty-one consecutive patients (4 women; mean age, 42 years) with fistula-in-ano were evaluated by local physical examination (inspection, probing, and digital examination), conventional ultrasound, and hydrogen peroxide-enhanced ultrasound before surgery. Ultrasound was performed using a B&K Diagnostic Ultrasound System with a 7-MHz rotating endoprobe. Hydrogen peroxide (3%) was infused via a small catheter into the fistula. The results of physical examination, ultrasound, and hydrogen peroxide-enhanced ultrasound were compared with surgical data as the criterion standard. The additive value of standard ultrasound and hydrogen peroxide-enhanced ultrasound compared with physical examination was also determined. RESULTS: At surgery, 8 intersphincteric and 11 transsphincteric fistulas and 2 sinus tracts (without an internal opening) were found. During physical examination, probing was incomplete in 13 patients, the diagnosis being correct in the other 8 patients (38%) as a low (intersphincteric or transsphincteric) fistula. With conventional ultrasound, the assessment of fistula-in-ano was correct in 13 patients (62%); defects in one or both sphincters could also be found (n = 8). With hydrogen peroxide-enhanced ultrasound, the fistulous tract was classified correctly in 20 patients, the overall concordance with surgery being 95%. The internal opening was found at physical examination in 15 patients (71%), with hydrogen peroxide-enhanced ultrasound in 10 patients (48%), and during surgery in 19 patients (90%). Secondary extensions, confirmed during surgery, were found in five cases. In two patients, a secondary extension with hydrogen peroxide-enhanced ultrasound was not confirmed during surgery. Both patients developed a recurrent fistula. CONCLUSION: Hydrogen peroxide-enhanced ultrasound is superior to physical examination and standard ultrasound in delineating the anatomic course of perianal fistulas. It makes accurate preoperative assessment of the fistula possible and may be of value for the surgeon in planning therapeutic strategy. PMID- 9749500 TI - Comparative study between intra-anal sponge and needle electrode for electromyographic evaluation of constipated patients. AB - PURPOSE: The aim of this study was to compare the intra-anal sponge electrode with the conventional needle electrode for electromyography of the pelvic floor in constipated patients. MATERIALS AND METHODS: Forty consecutive patients (27 females) with a mean age of 64.3 (range, 15-87) years who had chronic constipation were prospectively evaluated for electromyographic evidence of nonrelaxation or paradoxical contraction of the puborectalis and external anal sphincter during simulated defecation. The soft intra-anal sponge electrode and then the concentric needle electrode were used in each patient as an internal control. Furthermore, in all patients, cinedefecography was used as an independent standard to confirm the diagnosis. Agreement was calculated using the kappa statistic. RESULTS: Confirmation of needle electromyography was noted in 19 of 20 patients (95 percent) who had sponge electromyographic evidence of paradoxical activity. Similarly, concurrence was noted in 19 of 20 patients (95 percent) with normal relaxation of the puborectalis observed with the sponge electrode. Agreement between needle and sponge electromyography was very good (kappa = 0.9), between needle electromyography and cinedefecography was fair (kappa = 0.4), and between sponge electromyography and cinedefecography was moderate (kappa = 0.5). Furthermore, needle electromyography was more painful in all patients compared with sponge electromyography. CONCLUSION: The soft sponge surface intra-anal electrode is an excellent alternative to the needle electrode for assessment of puborectalis activity in constipated patients. Sponge electromyography has the advantage of being as accurate as, but less painful than, needle electromyography. PMID- 9749501 TI - Bowel resection for intestinal endometriosis. AB - PURPOSE: The study contained herein was undertaken to evaluate which factors predict a good outcome following intestinal resection for endometriosis. METHODS: A retrospective analysis of all patients undergoing bowel resection for severe (American Fertility Society Stage IV) endometriosis at one institution between the years 1992 and 1996 was conducted using systematic chart review and follow-up by telephone interview. RESULTS: Twenty-nine patients were identified within the study period. The most frequent symptoms were pelvic pain, abdominal pain, rectal pain, and dysmenorrhea. Nearly all patients (93 percent) underwent low anterior resection of the rectum and distal sigmoid. Other intestinal procedures were appendectomy, terminal ileal resection, cecectomy, and sigmoid resection. Thirty four percent of patients had simultaneous total abdominal hysterectomy and bilateral salpingooophorectomy. Complete follow-up was obtained on 26 patients (90 percent; mean follow-up 22.6 (range, 8-63) months). All patients (100 percent) reported subjective improvement. Forty-six percent of patients were "cured" according to the prospectively applied definition (resolution of symptoms without need for further medical or surgical therapy). The only variable analyzed that was associated with "cure" was concomitant total abdominal hysterectomy and bilateral salpingooophorectomy (odds ratio, 12; 95 percent confidence interval, 1.8-81.7). This association remained significant after correcting for age and the presence of gastrointestinal symptoms. CONCLUSION: Intestinal resection can be performed safely in most women with severe endometriosis and bowel involvement, although many of these patients experience persistent or recurrent symptoms. Total abdominal hysterectomy and bilateral salpingooophorectomy at the time of bowel resection correlates with improved outcome. PMID- 9749502 TI - Sacral reflexes: physiology and clinical application. AB - Sacral reflexes consist of motor responses in the pelvic floor and sphincter muscles evoked by stimulation of sensory receptors in pelvic skin, anus, rectum, or pelvic viscera. These responses may be elicited by physical or electrical stimuli. They have been used in research studies of the pathophysiology of pelvic floor and anorectal disorders and many have been recommended for diagnostic use. These reflexes are described and discussed in this review. More rigorous evaluation of their value in the clinical assessment and care of patients with pelvic floor and sphincter disorders is required. Currently direct comparisons of the value of particular responses are generally not available, and few of these reflexes have proven validity for use in clinical diagnosis. PMID- 9749503 TI - Giant colonic diverticulum: report of four cases and review of the literature. AB - PURPOSE: Giant colonic diverticulum are rare, with 103 reported cases in 95 patients. The experience of any one surgeon is limited. We aimed to retrospectively review our experience and to review the literature on origin, pathology, and management of this rare and unusual problem. METHOD: Cases were identified by review of pathologic database and by computerized audit from three hospitals. RESULTS: Five giant colonic diverticulum were identified in four patients, and the pathology and management were reviewed. CONCLUSION: A definition and classification system of giant colonic diverticulum is suggested. Giant colonic diverticulum should be the universal term to cover all colonic diverticulum larger than 4 cm, and we suggest that there are two types based on histology. Literature review reveals 103 reported cases in 95 patients. Type I (87 percent) is a pseudodiverticulum, perhaps related to conventional diverticular disease, whereas Type II (13 percent) is a true diverticulum, which is probably a type of communicating cystic congenital duplication. These lesions tend to occur in the sigmoid colon (93 percent) and present with complications similar to conventional diverticular disease. In the presence of conventional diverticular disease, consideration should be given to anterior resection, and in the absence, diverticulectomy should be considered. PMID- 9749504 TI - Neurofibromatosis of the colon and rectum combined with other manifestations of von Recklinghausen's disease: report of a case. AB - Gastrointestinal involvement of neurofibromatosis occurs in as many as 25 percent of cases. Neurofibroma occurs most frequently in the stomach and jejunum, but the colon may also be involved. This condition is characterized by multiple submucosal neurofibromas of the gastrointestinal tract and cafe au lait pigmentation, bony abnormalities, and neurofibromas of both central and peripheral nerves. The lesions consist of an overgrowth of neural tissue along with other mesenchymal elements. Gastrointestinal neurofibromas may cause occult bleeding, luminal obstruction, or intussusception. Malignant transformation into neurofibrosarcoma is rare. We encountered a case of neurofibromatosis diffusely involving the left colon, the sigmoid colon, and the rectum, which resulted in herniation of the mass through the anus, with intestinal obstruction. The patient also showed skin lesions of the neurofibromatosis. We report this case with a review of the literature. PMID- 9749505 TI - Cellular neurilemoma (schwannoma) of the descending colon mimicking carcinoma: report of a case. AB - We report a rare case of a cellular neurilemoma (schwannoma) of the descending colon, mimicking carcinoma, not accompanied by von Recklinghausen's disease. The differential diagnostic problems are discussed and the possibility of a site specific, modified Schwann cell of myenteric plexus origin is suggested. PMID- 9749506 TI - Usefulness of bipolar scissors for total colectomy. AB - PURPOSE: Total colectomy is frequently used in colorectal surgery. We evaluated the usefulness of bipolar scissors in this operation. METHODS: Twenty patients with ulcerative colitis underwent total colectomy at Hyogo College Hospital. Bipolar scissors were used in ten randomly chosen colectomies. The resected major vessels of the colonic mesentery were stained by hematoxylin and eosin. The bursting pressure of the artery weld was measured with a water-filled, open-tip catheter. RESULTS: Both number of ligations and operating time were reduced by use of bipolar scissors. The cut edge of the major artery was occluded by the weld created, with a mean bursting pressure of 213 (range, 175-250) mmHg. CONCLUSION: Use of bipolar scissors was safe and had the advantage of shortening the operating time by decreasing the number of clamping and ligating procedures. PMID- 9749507 TI - Artificial anal sphincter. PMID- 9749508 TI - Dorsolateral subluxation of hip joints in dogs measured in a weight-bearing position with radiography and computed tomography. AB - OBJECTIVE: develop a radiographic procedure to measure dorsolateral subluxation (DLS) of the femoral head in canine coxofemoral (hip) joints in a weight-bearing position. STUDY DESIGN: DLS measured on a radiographic projection was compared with DLS measured on computed tomography (CT) images of hip joints in a weight bearing position. ANIMALS: A total of 24 dogs of varying ages were examined including Labrador retrievers, greyhounds, and Labrador-greyhound crossbreeds. METHODS: Anesthetized dogs were placed in sternal recumbency in a kneeling position in a foam rubber mold. The stifles were flexed and adducted with the femora perpendicular to, and in contact with, the table. To test for DLS, dogs were imaged in this weight-bearing position (DLS test) with routine radiography and CT. For each hip, the DLS score was determined by measuring the percentage of the femoral head medial to the lateralmost point of the cranial acetabular rim on the dorsoventral radiographic projection and the lateralmost point of the central, dorsal acetabular rim on the CT image. Higher DLS scores indicated better coverage of the femoral head by the acetabulum. DLS scores were compared with the distraction index (DI) by grouping joints according to their probability of developing osteoarthritis (OA) as predicted by the DI. RESULTS: The DLS score in the new position ranged from 29% to 71% for radiography and 15% to 59% for CT. Joints classified as OA unsusceptible had a mean score of 64% +/- 1.5% for radiography and 55% +/- 0.8% for CT (n = 10); hip joints having a high probability of developing OA had a score of 39% +/- 2.6% for radiography and 26% +/- 1.9% for CT (n = 8). When the DLS test was repeated on the same dogs at a different time, the intraclass correlation coefficient for the DLS score on the radiographs was 0.85 (left hip) and 0.89 (right hip). There was a strong correlation (r = .89 for both hips) between the DLS score measured on the weight bearing radiograph and the CT image. A strong correlation also was observed between the DLS score and the DI (r = -.87). The DLS scores for OA unsusceptible joints and joints with a high probability of developing OA were significantly different (P < .05). CONCLUSIONS AND CLINICAL RELEVANCE: The DLS test can be performed with CT or routine radiography to measure variable amounts of DLS in weight-bearing hip joints oriented similarly to those of a standing dog. After additional long-term follow-up studies evaluating the development of OA and breed effects are performed, the DLS method may prove useful in studies of normal and abnormal hip joint development related to canine hip dysplasia. PMID- 9749509 TI - Microvascular free tissue transfer: results in 57 consecutive cases. AB - OBJECTIVE: To evaluate the outcomes and complications in a consecutive series of animals undergoing microvascular reconstructive procedures at two veterinary institutions. STUDY DESIGN: Retrospective study. ANIMALS OR SAMPLE POPULATION: A total of 44 client-owned dogs and one red-necked wallaby. METHODS: The medical records of all animals undergoing reconstructive microsurgical procedures at the Western College of Veterinary Medicine and Michigan State University were reviewed. Microvascular flap survival and related complications were described. Statistical analysis was performed to determine the significance of relationships between operative factors and outcome. RESULTS: A total of 57 microvascular procedures were performed on 55 animals. Reconstruction was required after trauma in 42 animals, after ablative cancer surgery in 11 animals and for correction of congenital tissue aplasia in I animal. Donor tissues included the superficial cervical cutaneous, medial saphenous fasciocutaneous or musculofasciocutaneous, caudal superficial epigastric cutaneous, trapezius muscle or musculocutaneous, caudal sartorius muscle, latissimus dorsi muscle or musculocutaneous, cranial abdominal myoperitoneal, carpal footpad, digital footpad, and vascularized ulnar bone flaps. A total of 53 of 57 flaps (93%) survived. There was a significant relationship between flap failure and level of assistant surgeon experience (P < .05). Latissimus dorsi flaps were significantly more likely to fail when compared with pooled data from all other flap types (P < .01). CONCLUSIONS: The success of microvascular tissue transfer in this case series compares favorably with those reported in human reconstructive microsurgery. Both the primary and assistant surgeon should be practiced in microsurgical technique. Failure of latissimus dorsi flaps was not likely caused by an inherently deficient flap design, but was more likely attributed to the location and severity of trauma at the recipient site, the difficulty in isolating suitable recipient vessels for anastomosis or the absence of a trained assistant surgeon during these procedures. Clinical Relevance-This retrospective study documents the successful application of microvascular technique in a series of clinical cases requiring tissue reconstruction. PMID- 9749510 TI - Complete ligation of extrahepatic congenital portosystemic shunts in nonencephalopathic dogs. AB - OBJECTIVE: To evaluate lack of encephalopathy as a positive prognostic factor for complete ligation of extrahepatic congenital portosystemic shunts in dogs. STUDY DESIGN: Retrospective analysis of case records. ANIMALS: Dogs with extrahepatic congenital portosystemic shunts treated at the Veterinary Medical Teaching Hospital of the College of Veterinary Medicine, Cornell University, from 1985 to 1996. METHODS: The ability to completely ligate the shunting vessel in 12 nonencephalopathic dogs was compared with that in 44 encephalopathic dogs with similar shunts. RESULTS: Clinical signs in the 12 nonencephalopathic dogs were related to ammonium biurate urolithiasis. All 12 dogs had single extrahepatic shunting vessels. The rate of complete ligation in the nonencephalopathic dogs was 92%, whereas the rate of complete ligation in the 44 encephalopathic dogs with single extrahepatic shunts was 59%. The ability to completely ligate the shunt in nonencephalopathic dogs was significantly better (P = .04) than in the encephalopathic dogs. CONCLUSION: Lack of encephalopathy is a positive prognostic factor for complete ligation of single extrahepatic congenital portosystemic shunts. CLINICAL RELEVANCE: In most affected dogs, extrahepatic congenital portosystemic shunts in nonencephalopathic dogs can be completely ligated. PMID- 9749511 TI - Limb salvage surgery for osteosarcoma of the proximal humerus: outcome in 17 dogs. AB - OBJECTIVE: To describe function and identify factors that affect outcome in dogs undergoing limb salvage surgery for osteosarcoma (OS) of the proximal humerus. STUDY DESIGN: A retrospective study of dogs in which OS of the proximal humerus was treated with limb salvage surgery. ANIMALS: 17 client-owned dogs. METHODS: Records were analyzed for functional outcome, recurrence, metastasis, and survival. RESULTS: Outcome was good to excellent in 12% of dogs. Recurrence, metastasis, and survival were significantly affected by completeness of surgical margins. Double plating of the distal allograft-host junction significantly reduced frequency of biomechanical failure. CONCLUSIONS: Limb salvage surgery for OS of the proximal humerus did not result in acceptable function and was fraught with postoperative complications. Outcome was significantly affected by completeness of surgical margins. CLINICAL RELEVANCE: Limb salvage surgery for OS of the proximal humerus in dogs cannot be recommended until improvement in functional outcome and reduction in postoperative complications can be achieved. The dependence of outcome on completeness of surgical margins supports aggressive en bloc resection and marking and evaluating surgical margins. PMID- 9749512 TI - Biomechanical comparison of the trocar tip point and the hollow ground tip point for smooth external skeletal fixation pins. AB - OBJECTIVE: To compare the insertional characteristics of external fixator pins with hollow ground (HG), modified HG, and trocar (T) points. STUDY DESIGN: An acute, in vitro biomechanical evaluation. SAMPLE POPULATION: Thirteen radii from canine cadavers. METHODS: A total of 16 T-tipped and 16 HG-tipped pins were inserted into 8 canine radii. Ten pins of each modification of the HG tip (length of the cutting edge reduced by 0.127 mm and 0.254 mm, respectively) were inserted into another five radii. All pins were inserted with low-speed power drilling and 80 N drilling load. Differences between peak tip temperature, drilling energy, and pullout force were determined for each pin type at both diaphyseal and metaphyseal locations. RESULTS: HG-tipped pins showed a 40% lower tip temperature in diaphyseal bone, a 25% reduction in drilling energy in diaphyseal bone, and a reduction of pullout force in both diaphyseal (65%) and metaphyseal (50%) bone compared with T-tipped pins. HG 0.254-mm pins generated higher tip temperatures and had greater pullout than HG pins in diaphyseal bone. CONCLUSIONS: The HG tip was a more efficient design; however, the reduction in pullout force suggests that, because a better hole was drilled, radial preload is reduced. Reduction of the cutting edge by 0.254 mm increased the pullout force but also increased the temperatures. CLINICAL RELEVANCE: Thermal and microstructural damage are reduced by the HG tip, but pin-bone interface stability is also compromised. The use of a tip with 0.254 mm reduction in the cutting edge may optimize the biological and mechanical factors at the pin-bone interface. PMID- 9749513 TI - A biomechanical evaluation and assessment of the accuracy of reduction of two methods of acetabular osteotomy fixation in dogs. AB - OBJECTIVE: To compare the accuracy of reduction, biomechanical characteristics, and mode of failure of two methods of acetabular osteotomy repair. STUDY DESIGN: Acetabular osteotomies were created in 16 paired hemipelves and stabilized with a screw/wire/polymethylmethacrylate composite fixation technique (SWP) or a 2-mm veterinary acetabular plate (VAP). Eight intact hemipelves were used as controls. SAMPLE POPULATION: Twelve canine cadavers. METHODS: Accuracy of osteotomy reduction was evaluated grossly and by measurement of articular incongruencies formed in polyvinylsiloxane impression casts. Acetabula were loaded in modified bending until failure using a universal testing machine. Data from load deformation curves were used to determine the biomechanical characteristics of the repaired and intact acetabula. Mode of failure was evaluated grossly and radiographically. RESULTS: Osteotomy reduction was superior in acetabula stabilized with SWP. Mean values +/- standard deviation for load at failure and stiffness of the intact acetabula were 2,796 +/- 152.9 N and 267.5 +/- 61.9 N/mm. Corresponding values for SWP and VAP were 1,192 +/- 202.7 N and 136.3 +/- 76.5 N/mm, and 1.100.5 +/- 331.6 N and 110.0 +/- 51.3 N/mm, respectively. The mean load at failure and stiffness of intact acetabula was significantly greater than acetabula stabilized with SWP or VAP. There was no significant difference between SWP and VAP for load at failure or stiffness. Failure of acetabula stabilized with SWP occurred by fracture of the polymethylmethacrylate and ventrolateral bending of the wires. Acetabula stabilized with VAP failed by ventrolateral twisting of the plate and bending of the caudal screws. CONCLUSIONS: SWP and VAP provide comparable rigidity, however, the SWP facilitates more accurate osteotomy reduction. CLINICAL RELEVANCE: These findings support the use of the SWP technique as an alternative method of acetabular fracture repair. PMID- 9749514 TI - Effect of dimethylsulfoxide on articular cartilage proteoglycan synthesis and degradation, chondrocyte viability, and matrix water content. AB - OBJECTIVE: To determine the effects of dimethylsulfoxide (DMSO) exposure on cartilage proteoglycan (PG) synthesis, PG degradation, chondrocyte viability, and matrix water content. STUDY DESIGN: Using a cartilage explant culture system, PG synthesis, PG degradation, matrix water content, and chondrocyte viability were determined for cartilage exposed to DMSO daily for selected periods of time. ANIMALS OR SAMPLE POPULATION: Juvenile bovine (calf) carpometacarpal joint cartilage explants. METHODS: PG synthesis: Explants (n = 30/group) were separated into 10 groups based on the time of daily exposure to 10% DMSO. Exposure time was repeated daily for 3 days. The control group was incubated in basal medium alone for 3 days, with daily medium changes. Once all DMSO exposure times were complete for the third day, PG synthesis was determined by analysis of incorporation of radiolabelled sulfate. Cell viability: Explants (n = 3/group) were subjected to an identical DMSO exposure protocol, and examined histologically. The percentage of viable cells/high power field (hpf) was calculated for each group. PG degradation: Explants (n = 21/group) were preincubated with radiolabelled sulfate, then subjected to a similar DMSO exposure protocol. The medium was collected from all explants daily and assayed for PG content. After 3 days, the explants were digested and total labelled PG content determined. Percent of total explant labelled PG content released into the medium daily was determined for each group. Water content: Explants (n = 21/group) were separated into three treatment groups, one of which had no treatments performed, whereas the other two groups were incubated in basal medium for 72 hours, one with, and one without, 10% DMSO. Wet and dry weights were determined, and percent water calculated, for all three groups. Separate 1-way ANOVA were performed, with appropriate post hoc tests (P < .05). RESULTS: PG synthesis was significantly lower than control for all time periods of DMSO exposure except for 1 and 3 hours, and decreased in a time-dependent manner after the 1-hour exposure time. The mean percentage of viable cells/hpf was significantly lower than control for the 1-, 3-, 9-, 12-, and 24-hour treatment groups. There was no significant difference in PG degradation for any group compared with control for the first 2 days of incubation. All groups except the 24-hour group had a significantly higher degradation compared with control for the third day of incubation. Cartilage exposed to DMSO for 72 hours had a significantly lower water content, and cartilage incubated in basal medium alone for 72 hours had a significantly higher water content than cartilage that received no DMSO and no incubation. CONCLUSIONS: DMSO, in relatively low concentration, is detrimental to articular cartilage PG synthesis in a time-dependent manner. Dehydration of the cartilage and chondrocyte death also occur with increasing time of DMSO exposure. Significant PG degradation occurs on the third day of culture with daily DMSO exposure. CLINICAL RELEVANCE: As a joint lavage solution, DMSO has potentially deleterious effects on the metabolism of chondrocytes. PMID- 9749515 TI - Repair of complete dorsal fracture of the proximal phalanx in two horses. AB - Simple complete dorsal fractures of the proximal phalanx were repaired in 2 mature pleasure horses with cortical bone screws placed in lag fashion. Healing occurred within 12 weeks and both horses returned to their previous performance level of light pleasure riding within 6 months of injury. PMID- 9749516 TI - Complications of balloon catheterization associated with aberrant cerebral arterial anatomy in a horse with guttural pouch mycosis. AB - A 3-year-old Quarter Horse gelding was treated for left guttural pouch mycosis by ligation and balloon catheterization of the left internal carotid artery. Catheter advancement was shorter (10 cm) than the normally reported distance (13 15 cm), but was observed endoscopically during placement as it coursed within the internal carotid artery through the guttural pouch. The horse developed a persistently abnormal respiratory pattern after catheter placement, failed to gain consciousness, developed pulmonary edema, and died 5.5 hours postoperatively. Postmortem examination revealed an aberrant left internal carotid arterial course with location of the embolectomy catheter at the junction of the basilar and caudal cerebellar arteries. Brainstem neuronal necrosis and alveolar and interstitial pulmonary edema were identified on histological examination. Angiography may be used to identify aberrant branching patterns. Failure to identify and occlude aberrant branches may result in fatal epistaxis and Brainstem lesions. PMID- 9749517 TI - Biomechanics of circular external skeletal fixation. PMID- 9749518 TI - Antinociceptive effects of oxymorphone-butorphanol-acepromazine combination in cats. AB - OBJECTIVE: To determine the antinociceptive effects of oxymorphone, butorphanol, and acepromazine individually and in combination to a noxious visceral stimulus in cats. STUDY DESIGN: Randomized, blinded controlled study. ANIMALS: Eight healthy mixed-breed cats (four male, four female) weighing 4.4 +/- 1.2 kg and aged 1 to 2 years old. METHODS: A silastic balloon catheter was inserted per rectum and inflated at various pressures. Physiological parameters (respiratory rate, pulse rate, and blood pressure) were also recorded. Subjects were administered individual and combined intravenous (i.v.) doses of 0.025, 0.05, 0.10, and 0.20 mg/kg oxymorphone and 0.025, 0.05, 0.10, and 0.20 mg/kg butorphanol. A further study of various ratios of butorphanol and oxymorphone (3:1, 2:1, 1:1, 1:2, and 1:3), at a combined equivalent dose of 0.1 mg/kg, was performed in four cats per dose combination. In a separate study, four cats were administered combined i.v. doses of 0.05 mg/kg each of oxymorphone and butorphanol or 0.05 mg/kg each of oxymorphone, butorphanol, and acepromazine. RESULTS: Combined doses of 0.05 and 0.10 mg/kg of oxymorphone and butorphanol showed mainly additive with some synergistic antinociceptive interactions and the combined dose of 0.2 mg/kg of each agent demonstrated additional antinociceptive effects, P < .05. Additional studies showed that various ratios of the two agents at a total combined dose of 0.10 mg/kg i.v. did not produce levels of antinociception that were significantly different from each other, P > .05. Acepromazine (ACE) significantly increased the magnitude of antinociception at 15 minutes when administered in combination with oxymorphone and butorphanol, P < .05. Also, physiological variables were unaffected by these drug combinations. CONCLUSIONS: Low doses of oxymorphone and butorphanol in combination can produce greater levels of antinociception than when used individually. ACE, in conjunction with oxymorphone and butorphanol, produced even greater levels of antinociception than the two-opioid drug combination. CLINICAL RELEVANCE: Oxymorphone, butorphanol, and ACE can be used in combination to produce additive or synergistic effects without adverse effects in cats. These data suggest that ACE and butorphanol at low doses given as preanesthetic medication followed by a mu opioid (eg, oxymorphone) after surgery at low doses may provide an effective method of pain management in the cat. PMID- 9749519 TI - Respiratory depressant and skeletal muscle relaxant effects of low-dose pancuronium bromide in spontaneously breathing, isoflurane-anesthetized dogs. AB - OBJECTIVE: To assess and compare the respiratory depressant and skeletal muscle relaxant effects of two low doses of a nondepolarizing neuromuscular blocker, pancuronium bromide. To determine if a "low dose" of pancuronium bromide can produce selective skeletal muscle relaxation in extraocular muscles sufficient to perform intraocular surgery while sparing or minimizing depression of muscles of ventilation. STUDY DESIGN: Blinded, randomized crossover, placebo controlled study. ANIMALS: Six healthy, adult mongrel dogs weighing 20.8 +/- 1.9 kg. METHODS: Spontaneously breathing, isoflurane-anesthetized dogs received 0.02 mg/kg pancuronium bromide, intravenously (i.v.), (high dose [HD]), 0.01 mg/kg pancuronium bromide, i.v., (low dose [LD]), or saline placebo i.v. in a blinded, randomized crossover study. Indices of patient ventilation including tidal volume (Vt), respiratory rate (RR), and minute ventilation (VE) were recorded throughout the study period. Serial arterial blood gas analyses were performed at timed intervals. Neuromuscular blockade of skeletal muscle was assessed at timed intervals with train-of-four stimulus/response ratios. Eye position scores, based on the degree of ocular rotation from a neutral gaze axis, were assigned by an ophthalmologist who was blinded to the treatment given. RESULTS: VT and VE in HD dogs decreased by 82% from baseline after administration of pancuronium bromide. Similarly, Vt and VE in LD dogs decreased 40% and 55%, respectively. Decreased ventilation in HD dogs corresponded with significant (P< .05) neuromuscular blockade, as indicated by train-of-four ratio less than 75% between 0 and 60 minutes. Eye position scores in HD and LD dogs were suitable for intraocular surgery between 0 and 60 minutes. Eye position scores in five of six control dogs were unsuitable for intraocular surgery at any time period. CONCLUSIONS: LD dogs experienced only transient, mild to moderate respiratory depression compared with HD dogs, which experienced prolonged, moderate to severe respiratory depression. Both LD and HD dogs acquired and maintained eye position scores suitable for intraocular surgery between 0 to 60 minutes. A "low dose" of pancuronium bromide, which would provide adequate extraocular muscle relaxation while minimizing ventilatory depression, was not identified. CLINICAL RELEVANCE: All patients receiving any dose of neuromuscular blocking agents should be closely monitored and receive ventilatory assistance as needed. PMID- 9749520 TI - Recovery from sevoflurane anesthesia in horses: comparison to isoflurane and effect of postmedication with xylazine. AB - OBJECTIVE: To compare recovery from sevoflurane or isoflurane anesthesia in horses. STUDY DESIGN: Prospective, randomized cross-over design. ANIMALS: Nine Arabian horses (3 mares, 3 geldings, and 3 stallions) weighing 318 to 409 kg, 4 to 20 years old. METHODS: Horses were anesthetized on three occasions with xylazine (1.1 mg/kg), Diazepam (0.03 mg/kg intravenously [i.v.]), and ketamine (2.2 mg/kg i.v.). After intubation, they were maintained with isoflurane or sevoflurane for 90 minutes. On a third occasion, horses were maintained with sevoflurane and given xylazine (0.1 mg/kg i.v.) when the vaporizer was turned off. Horses were not assisted in recovery and all recoveries were videotaped. Time to extubation, first movement, sternal, and standing were recorded as was the number of attempts required to stand. Recoveries were scored on a 1 to 6 scoring system (1 = best, 6 = worst) by the investigators, and by three evaluators who were blinded to the treatments the horses received. These blinded evaluators assessed the degree of ataxia present at 10 minutes after each horse stood, and recorded the time at which they judged the horse to be ready to leave the recovery stall. RESULTS: Mean times (+/- SD) to extubation, first movement, sternal, and standing were 4.1 (1.7), 6.7 (1.9), 12.6 (4.6), and 17.4 (7.2) minutes with isoflurane; 3.4 (0.8), 6.6 (3.1), 10.3 (3.1), and 13.9 (3.0) minutes with sevoflurane; and 4.0 (1.2), 9.1 (3.3), 13.8 (6.5), and 18.0 (7.1) with sevoflurane followed by xylazine. Horses required a mean number of 4 (2.3), 2 (0.9), and 2 ( 1.6) attempts to stand with isoflurane, sevoflurane, and sevoflurane followed by xylazine respectively. The mean recovery score (SD) for isoflurane was 2.9 (1.2) from investigators and 2.4 (1.1) from blinded evaluators. For sevoflurane, the mean recovery score was 1.7 (0.9) from investigators and 1.9 (1.1) from evaluators, whereas the recoveries from sevoflurane with xylazine treatment were scored as 1.7 (1.2) from investigators and 1.7 (1.0) from blinded evaluators. CONCLUSIONS: Recoveries appeared to vary widely from horse to horse, but were significantly shorter with sevoflurane than isoflurane, although sevoflurane followed by xylazine was no different from isoflurane. Under the conditions of the study, recoveries from sevoflurane and sevoflurane followed by xylazine were of better quality than those from isoflurane. CLINICAL RELEVANCE: Sevoflurane anesthesia in horses may contribute to a shorter, safer recovery from anesthesia. PMID- 9749522 TI - Lowering the pH optimum of D-xylose isomerase: the effect of mutations of the negatively charged residues. AB - Streptomyces rubiginosus D-xylose isomerase catalyzes the reversible isomerization of D-glucose to D-fructose. The isomerization reaction is maximized in the alkaline region of pH 8.5-8.8. The amino acid residues around two active site histidines (His-54 and His-220) and on the surface of the enzyme were mutated to improve the catalytic efficiency at neutral pH. The mutations have been made by removing the negatively charged residues based upon the sequence comparison of other D-xylose isomerases and the hypothesis proposed by Russell and Fersht (1987). The effects of these substitutions on kinetic parameter, pH dependence, and thermostability were characterized. The kcat values for D56N and E221A mutants on D-glucose are increased by 30-40% over that of the wild-type enzyme at pH 7.3 and the increased activities are maintained between pH 6 and 7.5. However, the surface mutants D65A, D81A, and D163N/E167Q only show 40-60% of the wild-type activity over the entire pH range. The pH activity profiles of the mutants are broader than that of the wild-type enzyme. The optimum pHs and the pKa values for all the mutants are lowered by 0.5-0.8 and 0.1-0.5 units, respectively. The small delta(deltaG) and high Tm values for all the mutants indicate that there is no significant change in the hydrogen bond network in the active site by mutations. These results indicate that D56N and E221A are possible candidates as good catalysts for High-Fructose Corn Syrup (HFCS) production. PMID- 9749521 TI - Plasma concentrations and cardiovascular influence of lidocaine infusions during isoflurane anesthesia in healthy dogs and dogs with subaortic stenosis. AB - OBJECTIVE: To determine the plasma concentrations and cardiovascular changes that occur in healthy dogs and dogs with aortic stenosis that are given an infusion of lidocaine during isoflurane anesthesia. STUDY DESIGN: Phase 1, controlled randomized cross-over trial; Phase 2, before and after trial ANIMALS: Phase 1, 6 healthy dogs (4 female, 2 male) weighing 23.8 +/- 7.4 kg; Phase 2, 7 dogs (4 female, 3 male) with moderate to severe subaortic stenosis (confirmed by Doppler echocardiography) weighing 31.1 +/- 14.5 kg. METHODS: After mask induction, intubation, and institution of positive pressure ventilation, instrumentation was performed to measure hemodynamic variables. After baseline, measurement at an end tidal isoflurane concentration of 1.9% (phase 1) or 1.85% (phase 2), a loading dose infusion of lidocaine at 400 microg/kg/min was given. Phase 1: Maintenance doses of lidocaine were administered consecutively (40, 120, and 200 microg/kg/min) after the loading dose (given for 10, 10, and 5 minutes, respectively) in advance of each maintenance concentrations. Measurements were taken at the end of each loading dose and at 25 and 35 minutes during each maintenance level. The same animals on a different day were given dextrose 5% and acted as the control. Phase 2: Dogs were studied on a single occasion during an infusion of lidocaine at 120 microg/kg/ min given after the loading dose (10 minutes). Measurements occurred after the loading dose and at 25 and 35 minutes. A blood sample for lidocaine concentration was taken at 70 minutes. Data were compared using a one-way ANOVA for phase 1, and between phase 1 and 2. Statistical analysis for phase 2 was performed using a paired t-test with a Bonferroni correction. A P value < or = .05 was considered significant. RESULTS: Phase 1: Plasma lidocaine concentrations achieved with 40, 120, and 200 microg of lidocaine/kg/min were 2.70, 5.27, and 7.17 microg/mL, respectively. A significant increase in heart rate (HR) (all concentrations), central venous pressure (CVP), mean pulmonary arterial pressure (PAP), and a decrease in stroke index (SI) (200 microg/kg/min) were observed. An increase in systemic vascular resistance (SVR) and mean PAP, and a decrease in SI also followed the loading dose given before the 200 microg/kg/min infusion. No other significant differences from the control measurements, during dextrose 5% infusion alone, were detected. Phase 2: Plasma lidocaine concentrations achieved were 5.35, 4.23, 4.23, and 5.60 microg/mL at 10, 25, 35, and 70 minutes, respectively. They were not significantly different from concentrations found in our healthy dogs at the same infusions. A significant but small increase in CVP compared with baseline was noted after the loading dose. There were no significant differences from baseline shown in all other cardiovascular data. There were no statistically significant differences in any measurements taken during the lidocaine infusion between the dogs in phase 1 and phase 2. Dogs with aortic stenosis tended to have a lower cardiac index than healthy dogs at baseline (88 v 121 mL/kg/min) and during lidocaine infusion (81 v 111 mL/kg/min). A small, statistically significant difference in systolic PAP was present at baseline. CONCLUSIONS: There does not appear to be any detrimental cardiovascular effects related to an infusion of lidocaine at 120 microg/kg/min during isoflurane anesthesia in healthy dogs or dogs with aortic stenosis. The technique used in this study resulted in therapeutic plasma concentrations of lidocaine. CLINICAL RELEVANCE: Methods shown in the study can be used in clinical cases to achieve therapeutic lidocaine levels without significant cardiovascular depression during isoflurane anesthesia. PMID- 9749523 TI - Tissue-specific expression and activation of N-methylpurine-DNA glycosylase in thymic carcinomas of transgenic mice expressing the SV40 large T-antigen gene. AB - Simian virus 40 T-tumor antigen (SV40 T-ag) can induce a wide variety of tumors in hamsters and neonatal mice. These tumorigenic effects are predominantly due to the activity of early viral gene products, large T-antigen and small t-antigen. We have analyzed the expression of a DNA repair gene, N-methylpurine-DNA glycosylase (MPG), from different tissues of a non-transgenic (control) and SV40 T-ag expressing transgenic mice at the mRNA level. Expression of the transgene in thymus of adult mice was also detected by the presence of SV40 T-ag mRNA. Non transgenic mice did not express the SV40 T-ag gene in their thymus, while the mRNA for MPG was found in thymus from both of transgenic and non-transgenic mice. The MPG gene was expressed in various tissues and is regulated in a tissue specific manner. Northern blot analysis revealed that the transgenic mice showed considerably higher expression of MPG in the thymic carcinomas. The level of MPG mRNA in the thymic carcinoma was elevated about 5.7 fold, as compared with those found in the control thymus. MPG expression was significantly increased, either directly or indirectly, by the SV40 T-ag gene product. These findings provide the first in vivo observations that the SV40 T-ag gene induced thymic carcinomas associated with the activation of the DNA repair gene, MPG. PMID- 9749524 TI - Coordinated expression of defense-related genes by TMV infection or salicylic acid treatment in tobacco. AB - To understand the coordinated functions of the different classes of defense related genes expressed in plant disease resistance, the expression patterns of pathogenesis related (PR) protein genes and genes involved in antioxidation and the production of secondary metabolites were examined. The expression patterns of the respective defense-related genes were monitored following TMV infection or salicylic acid treatment. Northern blot analyses showed that PR genes such as PR 1, beta-1,3-glucanase and chitinase were strongly induced in tobacco leaves upon TMV infection or salicylic acid treatment. 3-Hydroxy-3-methylglutaryl CoA reductase (HMGR) and phenylalanine ammonialyase (PAL), involved in isoprenoid and phenylpropanoid biosynthesis, respectively, were mildly induced at the late stage of normal hypersensitive response (HR) or after salicylic acid treatment when compared with the PR-gene expressions. However, in acute HR, they were strongly expressed at the early stage. Interestingly, the expression of the antioxidative genes, anionic peroxidase and ascorbate peroxidase, were inversely expressed following TMV infection and salicylic acid treatment. Differential expression of 3 groups of genes involved in plant defense responses are discussed in relation to different signal transduction pathways. PMID- 9749525 TI - Molecular characterization of the virulence gene virG of pTiKU 12. AB - The virulence (vir) genes of Agrobacterium tumefaciens KU12, a Korean strain, were not induced by acetosyringone and the strain showed weak tumor forming ability and broad plant host ranges. We identified complete nucleotide sequence of virG of pTiKU12, an octopine Ti plasmid of this strain. When it was compared with those of other Ti plasmids, pTiKU12 virG contained an open reading frame (ORF) of 726 nucleotides which showed much lower homology (about 77%) than those (above 98%) already known among octopine Ti plasmids and it started with GTG codon instead of TTG found in other Ti plasmids. Only two vir boxes and one promoter region were confirmed in 5'-untranslated region instead of three vir boxes and two promoters which were found in pTiA6 virG. Nevertheless, important amino acids for the functional activity of VirG were so conserved that the virG included in pUCDG could complement a virG mutant Agrobacterium tumefaciens Mx19 in beta-galactosidase activity assays and on plant tumor tests. PMID- 9749526 TI - Exon-intron structure of the human PTK6 gene demonstrates that PTK6 constitutes a distinct family of non-receptor tyrosine kinase. AB - Partial PTK6 (also known as Brk) cDNA was initially isolated by reverse transcription-PCR of normal human melanocyte mRNAs and the full-length cDNA encodes a non-receptor protein tyrosine kinase with an SH3 domain, an SH2 domain, and a kinase catalytic domain. We have cloned the human PTK6 gene by screening human genomic lambda libraries using the full-length PTK6 cDNA as probe. The human PTK6 gene consists of 8 exons encompassing 8.8 kb and all the splicing junctions followed the conserved GT/AG rule. Coding sequence of the PTK6 gene was identical to that of the cDNA cloned from T-47D, human breast tumor cell line. Although the amino acid sequence of the PTK6 polypeptide showed the strongest homology to those of the Src family members of protein tyrosine kinases, exon intron boundaries of the PTK6 gene were quite different from those of the Src family genes, which are evolutionarily conserved. The 813-bp 5'-flanking sequence of the PTK6 gene upstream of a luciferase reporter gene conferred significant promoter activity, at approximately 60% level of the SV40 promoter, in transient expression assays into MCF-7, human breast tumor cell line. PTK6 mRNA was expressed at very high level in colon and at high levels in small intestine and prostate, and at low levels in some tested fetal tissues. These results suggest that PTK6 constitutes an evolutionarily distinct family of non-receptor protein tyrosine kinases and may function as an intracellular signal transducer in specific tissues. PMID- 9749527 TI - Induction of late activation events by Igbeta signaling subunit of B-cell receptor in Jurkat T-cell without tyrosine phosphorylation. AB - T-lymphocyte activation consists of multiple intracellular signaling events, eventually leading to cellular proliferation by the control of cytokine gene expression and the acquisition of diverse effector function. To investigate the functional specificity of ITAM (Immunoreceptor Tyrosine-based Activation Motif), chimeric molecules CD8-zeta, CD8-Igalpha, CD8-Igbeta, which contain the extracellular and transmembrane domains of the human CD8alpha molecule and the cytoplasmic tail of T-cell receptor (TcR) chain, Igalpha or Igbeta subunit of B cell receptor, respectively, were stably expressed in a Jurkat cell line. Upon stimulation with anti-CD8 mAb OKT8, CD8-zeta and CD8-Igalpha chimeric proteins induced tyrosine phosphorylation of various cytoplasmic substrates as seen in TcR stimulation. They were also capable of stimulating IL-2 gene expression in a NF AT dependent manner and inducing CD69 expression on the surface. However, stimulation of CD8-Igbeta can induce activation of CD69 surface expression and IL 2 gene expression equivalent to the level by CD8-Igalpha and CD8-zeta without induction of the tyrosine phosphorylation of intracellular signaling molecules. These results suggested that some of signaling chains containing ITAM may utilize a signal pathway without substrate tyrosine phosphorylation during T-cell activation leading to the IL-2 secretion. PMID- 9749528 TI - Overexpression and characterization of the cDNA encoding the coat protein of cucumber mosaic virus (Strain ABI) isolated in Korea. AB - We constructed a recombinant CMVCP expression vector termed pMALCMV in which cDNA fragment encoding CMVCP is ligated into pMAL-c2, an E. coli expression vector. Overexpression of pMALCMV containing the entire open reading frame of CMV cDNA sequence and the maltose binding protein (MBP) leader gene was facilitated in E. coli TB1 cells, which resulted in the production of a fusion protein of MBP-CMVCP (Mr 67.7 kDa) that was immunoprecipitable with rabbit polyclonal antiserum specific for MBP. The CMVCP (Mr 24.5 kDa) was isolated through a preparative SDS polyacrylamide gel following digestion of the affinity ligand purified fusion protein with Factor Xa. The partial amino acid sequences of the cleaved proteins were confirmed at the amino terminus by peptide sequencing. The CMVCP antiserum was also prepared by intraperitoneal injection of this purified CP into a BALB/c mouse. Immunoblot analysis showed that the purified CMVCP from the Factor Xa cleavage reaction was an authentic overexpression product of the cloned CMVCP. Using an RNA mobility shift assay, it was demonstrated that CMVCP can bind to its own RNA transcript in a concentration dependent manner. However, the complex formed between CMVCP and its RNA was abolished by the addition of a polyclonal antibody that had been raised against CMVCP, confirming that the overexpressed CMVCP specifically interacts with its own RNA. Thus, our results can provide a basis for the development of a hybridoma cell line expressing the monoclonal antibody for CMVCP and molecular cloning of their genes, which may lead to the creation of CMV-resistant transgenic plants. PMID- 9749529 TI - Topoisomerase II-mediated DNA cleavage on the cruciform structure formed within the 5'upstream region of the human beta-globin gene. AB - A 52 base pair alternating purine-pyrimidine (RY) repeat sequence lies in the 5' upstream region of the human beta-globin gene. The structural transition of a plasmid containing this repeat was analyzed by two-dimensional gel electrophoresis. These conformational studies indicate that the 52 bp RY repeat undergoes local transition from the right-handed B-DNA into a cruciform DNA under torsional stress and the transition initiates at a threshold level of negative supercoiling (-sigma = 0.042). The superhelicity-dependent S1 nuclease cleavage sites were mapped only within the RY repeat and no nicking was observed outside of the repeat. In view of the fact that DNA topoisomerase II is highly reactive towards RY repeat which can adopt unusual DNA conformation, we have investigated the effects of the superhelicity-dependent conformational transition of the 52 bp RY repeat on topoisomerase II cleavages. Cleavage reactions were performed on the pRYG plasmid with varying levels of negative superhelical densities ranging from 0 to -0.074. Under the low torsional stress, topoisomerase II cleavage activity at the RY repeat gradually increased with the increasing levels of negative superhelical densities. However, over a threshold level of negative supercoiling for cruciform conformation, the intensities of enzyme cleavage sites at the RY repeat were essentially identical. These results suggest that topoisomerase II can bind and cleave the cruciform structure in a dynamic process identical to duplex B-DNA. PMID- 9749530 TI - Thioltransferase from Schizosaccharomyces pombe: purification to homogeneity and some properties. AB - Two types of thioltransferase were identified in the cytosolic extract of Schizosaccharomyces pombe, a fission yeast. In the present study, the major one of them was purified to homogeneity using chromatography processes such as ion exchange chromatography and gel filtration. Purification was monitored by the transhydrogenase activity of thioltransferase with 2-hydroxyethyl disulfide as a substrate. Its molecular weight was estimated to be about 14,000 on SDS polyacrylamide gel electrophoresis. The purified enzyme catalyzes the reduction of various disulfide compounds such as S-sulfocysteine, L-cystine, and insulin. It was also found to contain the reducing activity on non-disulfide substrates such as dehydroascorbic acid and alloxan. Its activity was greatly activated by high concentrations of reduced glutathione. It was found to be very heat-stable as like other thioltransferases. It was characterized on other aspects such as kinetic parameters and optimal reaction conditions. PMID- 9749531 TI - Cloning, nucleotide sequence, and expression of the DNA ligase-encoding gene from Thermus filiformis. AB - The gene encoding Thermus filiformis (Tfi) DNA ligase was cloned and its nucleotide sequence was determined by the chain-termination method. The primary structure of Tfi DNA ligase was deduced from its nucleotide sequence. The Tfi DNA ligase comprises of 667 amino acid residues and its molecular mass was determined to be 75,936 Da. The deduced amino acid sequence of Tfi DNA ligase showed a 86.5% homology to Tth DNA ligase and 43.5% to E. coli DNA ligase. The Lys-116 of Lys Val-Asp-Gly motif was proposed to be the active residue of Tfi DNA ligase. In comparison with the amino acid composition of DNA ligase, Tfi DNA ligase showed a significant increase in the proportion of charged residues, Arg and Glu, compared to E. coli DNA ligase. The G + C content in the first, second, and third positions of the codons used were 70.3%, 40.3%, and 90.3%, respectively. Codon usage in Tfi DNA ligase was heavily biased towards the use of G + C in the third position. Under tac promoter control, Tfi DNA ligase was overproduced to greater than 9% of E. coli BL26Blue cellular proteins. PMID- 9749532 TI - Hepatitis C virus envelope DNA-based immunization elicits humoral and cellular immune responses. AB - The vaccine development for hepatitis C virus (HCV) is highly urgent to prevent non A and non B hepatitis. It was recently shown that the HCV envelope proteins appeared to the key viral antigens to induce protective immunity. To generate immune responses to the HCV envelope proteins on the DNA-based immunization, various envelope gene-containing plasmids were constructed. For efficient expression and secretion of envelope proteins, the signal sequence of each envelope protein was replaced with either herpes simplex virus type-1 (HSV-1) gD or signal sequence of gD and truncated C-terminal hydrophobic regions of envelope proteins. The intramuscular injection of these plasmids generated a significant level of antibody titers to the E1 and E2 proteins, which maximally reached 850 and 25,000 respectively. The secreted form of each envelope protein and the fusion of the highly immunogenic gD proteins were shown to have no significant effect on generating immune responses to the envelope proteins. In addition, immunized rats appeared to generate antibodies directed to the homologous HVR-1 peptide. Splenic lymphocytes from immunized rats were shown to induce significant T-cell proliferative responses with the stimulation of recombinant E1 and E2 proteins. Our results demonstrated that the HCV envelope-DNA based immunization could elicit both humoral and cellular immune responses. PMID- 9749534 TI - Major identity element of glutamine tRNAs from Bacillus subtilis and Escherichia coli in the reaction with B. subtilis glutamyl-tRNA synthetase. AB - Early investigations revealed that Bacillus subtilis glutamyl-tRNA synthetase [GluRS (bs)] is responsible for aminoacylating both glutamate tRNA [tRNA(Glu) (bs)] and glutamine tRNA [tRNA(Gln) (bs)] with glutamate. The same Bacillus enzyme can also efficiently attach glutamate to one isoacceptor glutamine tRNA [tRNA(Gln) (ec)] of Escherichia coli in vitro but not to tRNA2(Gln) (ec) and tRNA(Glu) (ec). To characterize identity elements of these glutamine tRNAs in the interaction with GluRS (bs), tRNA2(Gln) (ec), tRNA1(Gln) (ec), three other mutant glutamine tRNAs [tRNA2(Gln) (AU) (C34 --> U34), tRNA2(Gln) (12M) (C34 --> U34, 31A-U39 --> 31U-A39), and tRNA2(Gln) (M21) (64C --> G50 --> 64G-C50, 63U-A51 --> 63A-U51)] originated from tRNA2(Gln) (ec), tRNA(Gln) (bs), and a mutant tRNAM(Gln) (bs) whose U at the 34th position (U34), was replaced to C (C34), were produced in E. coli. All of the E. coli glutamine tRNAs containing U34 such as tRNA1(Gln), tRNA2(Gln) (AU), and tRNA2(Gln) (12M) could be charged with glutamate by GluRS (bs), whereas tRNA2(Gln) (ec) and its T-stem mutant tRNA2(Gln) (M21) containing C34 could not be charged by the same enzyme. The unique change of C34 to U34 of tRNA2(Gln) (ec) acquired glutamate acceptor activity by GluRS (bs). This result suggests that the U34 is the major identity element of tRNA1(Gln) (ec) in the recognition by GluRS (bs). The same situation was found in tRNA(Gln) (bs). The glutamate acceptor activity of tRNA(Gln) (bs) disappeared on replacement of U34 to C34. To find out whether modified bases in tRNA(Gln) (bs) are involved in the recognition by GluRS (bs), glutamylation of tRNA(Gln) (bs) produced by in vitro transcription was also examined but the in vitro transcript of tRNA(Gln) (bs) could not be charged with glutamic acid by GluRS (bs). All of these mean that the major recognition element for GluRS (bs) is U at the 34th position of both tRNA(Gln) (bs) and tRNA1(Gln) (ec) as a modified form. PMID- 9749533 TI - Expressed sequence tags of radish flower buds and characterization of a CONSTANS LIKE 1 gene. AB - Expressed sequence tag (EST) analysis was conducted for young flower buds of radish plants. Among a total of 66 ESTs examined, 40 showed a significant similarity to previously identified genes. Twenty-eight ESTs were similar to proteins identified in other plants, 11 were similar to eukaryotic proteins other than plants, and one was similar to a prokaryotic protein. Four clones were selected for further studies. EST clone 81, which showed a homology to germin like proteins was expressed more abundantly in leaves and roots as compared to flower buds. Clone 105 was highly homologous to the translation inhibitor protein and was expressed in all three organs, but the expression level was higher in flower buds and roots. Another EST clone, 133, which shared a significant similarity with the Ran-binding protein, hybridized to two different size transcripts that were detectable only in flower buds. Clone 39 was a homolog of CONSTANS, which is a gene involved in controlling the flowering time in Arabidopsis. The cDNA clone of EST clone 39 containing the entire open reading frame was obtained and designated as RsCOL1 (Raphanus sativus CONSTANS LIKE 1). It was 1049 bp long and contained an open reading frame of 307 amino acid residues (calculated molecular mass = 33.1 kDa). The RsCOL1 protein contained two putative zinc finger motifs in the amino terminal region which were 59% identical to the corresponding region of the Arabidopsis CO protein. The radish protein also contained a predicted nuclear localization domain in the carboxyl terminal region which was 87% identical to the corresponding region of CO. DNA blot analysis revealed that the radish genome contained several genes similar to RsCOL1. RNA blot analysis showed that RsCOL1 was strongly expressed in flower buds at the early bolting stage, and the expression level declined as the flower bud matured. The transcript was also detectable in leaves and roots. In mature flowers, the RsCOL1 transcript was present primarily in carpels. PMID- 9749535 TI - Cloning and characterization of plastid ribosomal protein S16 gene from potato (Solanum tuberosum L. cv Desiree). AB - The plastid ribosomal protein s16 (rps16) gene was cloned from potato (Solanum tuberosum L. ssp. tuberosum cv Desiree) by PCR amplification to obtain a new homologous recombination site of plastid transformation. The potato rps16 genomic clone was 1627 bp in size and the coding region was interrupted by an 859 bp intron. Exon I was 40 bp, encoding 13 amino acids and exon II was 227 bp, encoding a 76 amino acid polypeptide. The nucleotide sequence of the rps16 gene from the "Desiree" potato shared perfect identity with the sequence from the "Superior" potato in the coding region. Three nucleotide substitutions, two nucleotide insertions, and one nucleotide deletion were found between the intron sequence of both "Desiree" and "Superior" cultivars. The amino acid sequence of the potato rps16 gene showed a high level of identity with rice, maize, tobacco, and mustard (84-94%) and a relatively low level compared with Bacillus stearothermophilus and E. coli (27-28%). Expression of the rps16 gene was strong in chloroplasts and transcripts were detectable in amyloplasts, suggesting that the rps16 gene is active in nonphotosynthetic plastids as well as in photosynthetic plastids. These results indicate that the potato rps16 gene can be used as a new homologous recombination site of plastid transformation for potato cultivars. PMID- 9749537 TI - Expression, purification, and characterization of a familial amyotrophic lateral sclerosis-associated D90A Cu,Zn-superoxide dismutase mutant. AB - Cu,Zn-superoxide dismutase (SOD) is known to be a locus of mutation in familial amyotrophic lateral sclerosis (FALS). We cloned the FALS mutant, D90A, and wild type of human Cu,Zn-SOD, overexpressed them in E. coli, purified the proteins, and studied their properties. We investigated their enzymic activities for catalyzing the dismutation of superoxide anions and the generation of free radicals with H2O2 as a substrate. Our results showed that both wild-type and mutant enzymes have identical dismutation activities. However, the hydroxyl radical-generating function of the D90A mutant, as measured using a 2,2'-azinobis (3-ethylbenzthiazoline-6-sulfonate), was enhanced relative to that of the wild type enzyme. Catalysis of this reaction by D90A was more sensitive to inhibition by the copper chelators, penicillamine and diethyldithiocarbamate, than was catalysis by wild-type Cu,Zn-SOD. Our study suggests that this gain-of-function of FALS mutant may, in part, be responsible for the development of FALS symptoms. PMID- 9749536 TI - Characterization of bovine and human cDNAs encoding NAP-22 (22 kDa neuronal tissue-enriched acidic protein) homologs. AB - We have characterized the bovine and the human cDNAs encoding the NAP-22 (22 kDa neuronal tissue-enriched acidic protein) homologs. Both bovine and human cDNAs encode proteins of 227 amino acids. The deduced amino acid sequences of the bovine and the human proteins are 63% and 65% identical, respectively, to that of rat NAP-22 protein, strongly suggesting that both the cDNAs characterized encode NAP-22 proteins. They also share 45% and 41% amino acid sequence identities with chicken CAP-23 (23 kDa cytoskeleton associated protein). Several important protein motifs, including myristoylation and phosphorylation sites, are well conserved in sequences and positions in all three mammalian NAP-22 proteins and chicken CAP-23 proteins. The bovine cDNA was characterized further. Southern analysis of the bovine genomic DNA suggests that the bovine NAP-22 protein is encoded by a single-copy gene. RNA blot analysis revealed that the bovine gene for NAP-22 protein encodes a 1.7 kb transcript that is present only in the brain. Our data suggest that the four proteins, bovine and human NAP-22 homologs, rat NAP-22, and chicken CAP-23, have homologous functions in different organisms. PMID- 9749538 TI - Transcriptional control of the glnD gene is not dependent on nitrogen availability in Escherichia coli. AB - Glutamine synthetase (GS) is one of the most important enzymes in the assimilation of nitrogenous compounds in Escherichia coli and related bacteria. For the control of its activity and biosynthesis, tricyclic cascades of uridylylation/deuridylylation of PII protein, adenylylation/deadenylylation of glutamine synthetase, and phosphorylation/dephosphorylation of Ntr1 are operating, where the regulation of uridylylation/deuridylylation by uridylyl transferase-uridylyl removing enzyme (UT-UR) (the product of the glnD gene) would play the ultimate nitrogen sensing role. However, the possible nitrogen regulatable element in the upstream of the glnD gene has been debated. In the present experiment, we have cloned and sequenced the four minute regions of the Escherichia coli chromosome, where rpsB, map, glnD, and dapD genes have been identified in sequence. We could localize the transcriptional start site at seven nucleotides upstream of the translation initiation codon by primer extension analysis. The nitrogen dependency of the glnD gene has been analyzed by Northern blot, RNase protection, and promoter-luciferase activity assays. These data suggested a constitutive expression of the glnD gene independent of nitrogen availability. From these results, it could be concluded that the ultimate nitrogen sensing device for the bacteria should be the UT-UR itself, through modulation of its activity in response to the nitrogen status rather than its biosynthetic mechanism. PMID- 9749539 TI - Molecular cloning and characterization of a plastid chaperonin 60 alpha subunit from Canavalia lineata. AB - A 2040 bp cDNA encoding plastid chaperonin 60alpha protein was isolated from a cDNA expression library prepared from Canavalia lineata leaves. The sequence analysis of the clone showed a significant homology to those of other plant plastid chaperonin 60alpha. The genomic DNA blot analysis indicated that Canavalia lineata chaperonin 60alpha is encoded by a single gene in the genome. The short-term kinetics of light on the gene expression revealed that the remarkable accumulation of the gene occurred after 24 h. The mRNA was also detectable in dark-grown seedling leaves. PMID- 9749540 TI - Interaction of neurotrophins and retinoic acid in neuronal differentiation. PMID- 9749541 TI - Lateral laryngotomy for the removal of Teflon granuloma. AB - Teflon injection has been used for vocal fold medialization following paralysis. Recently, numerous articles have discussed the complications of Teflon injection, including overinjection, airway obstruction. Teflon granuloma, and an abnormal mass effect creating a decreased vibratory character of the true vocal fold. Multiple techniques for Teflon removal have been described. This report details our experience with complete Teflon granuloma removal via a lateral laryngotomy under local anesthesia. Microscopic dissection of the entire granuloma and the paraglottic space was accomplished in all patients. Due to extensive destruction caused by the granuloma, the vocal ligament was resected in 3 patients; it was partially resected and reanastomosed in 1 case, and spared in 6 patients. Laryngeal reconstruction was accomplished with an inferiorly based sternohyoid muscle rotation flap and arytenoid adduction. Effortful speech secondary to pressed vocal quality resolved in all patients. Near-normal to normal vocal quality was obtained in 4 patients, with the average "voice desirability" improving 60% and the effective glottic width increasing 29%. Factors that contributed to a successful outcome included noninvolvement of the vocal ligament and sparing of the mucosal cover. PMID- 9749542 TI - Meyer procedure for severe laryngotracheal stenosis. AB - Many surgical procedures have been devised to manage laryngotracheal stenosis secondary to trauma. Laryngotracheal atresia is the most severe form and the most difficult to repair. The Meyer procedure is a three-stage operation that provides structural support that is covered with mucosa. A laryngotracheal trough is created and a carved trough-shaped cartilage graft is placed above and lateral to it in the first stage. The skin over the graft is replaced by buccal mucosa in the second stage. In the last stage, the cartilage graft with overlying mucosa is swung onto the trough as a composite flap replacing the anterior and lateral laryngeal and tracheal walls. Attempt at reconstruction was made in 8 patients. All but one lesion was secondary to endotracheal intubation. Two patients were unable to be taken to completion of the third stage. Of the remaining 6 patients, all have a functional voice and only 1 remains cannulated at night. PMID- 9749543 TI - Recording from afferents in the intact recurrent laryngeal nerve during respiration and vocalization. AB - The goal of this study was to determine whether sensory fibers in an intact recurrent laryngeal nerve (RLN) are influenced by respiration or vocalization. Patterns of RLN afferent activity were examined during respiration and evoked vocalization by means of midbrain electrical stimulation in cats anesthetized with alpha-chloralose. Nerve bundles were dissected from an intact RLN, with motor function preserved. The bundles were cut and the laryngeal end was placed on floating bipolar electrodes. Fifteen right RLNs were examined. A total of 9 single and multiunit afferent fibers from 4 cats were isolated and examined during respiration. Four units, analyzed from 3 fibers, showed respiratory phase modulation. Eight units, analyzed from 4 fibers in 1 cat, were observed during vocalization and showed no vocalization phase modulation. The RLN afferents could contribute to reflex modulation of the respiratory cycle, but more extensive sampling would be necessary to preclude effects from vocalization. PMID- 9749544 TI - Effect of penicillin on formation of fibrous adhesions in acute otitis media. AB - Fibrous middle ear adhesions are occasionally encountered in middle ear surgery and may cause a hearing impairment. Although usually associated with chronic otitis media, adhesions are also found following a single episode of experimental acute suppurative otitis media, suggesting a pathogenesis based on the inflammatory process engaging acute infection. In a well-established rat model of pneumococcal acute otitis media, we report on the effect of penicillin V on formation of fibrous middle ear adhesions. Previous studies have shown marked impact of penicillin on mucosal goblet cell density and other histopathologic features. Number, anatomic localization, and histopathologic morphology of adhesions were assessed in a longitudinal study of 25 normal, 25 untreated, and 25 treated rats. Although penicillin administration induced a slight tendency toward fewer ears with adhesions and fewer adhesions per ear, these changes were nonsignificant. Histomorphology and the general pattern of anatomic localization of adhesions were unaffected by penicillin administration. We conclude that administration of penicillin has an inconspicuous effect on the formation of fibrous adhesions in experimental acute otitis media caused by Streptococcus pneumoniae. PMID- 9749545 TI - Mucocele of the sphenoid sinus: a late complication of transsphenoidal pituitary surgery. AB - In an 18-month period, 3 cases of sphenoidal mucocele following pituitary surgery were diagnosed at our institution. Only 1 case of this late-onset disorder has yet been reported as a pitfall of the transsphenoidal route. Symptoms include recurrent headache and visual complications. Diagnosis was delayed because of misinterpretation of the magnetic resonance imaging findings, which actually showed the development of a sphenoid mucocele long before clinical symptoms occurred. These 3 cases suggest that attention should be focused not only on the sella turcica, but also on the sphenoid sinus, in analyzing the magnetic resonance imaging data. The treatment consists of endoscopic transnasal marsupialization, since the mucocele is lined by normal epithelium with an inflammatory reaction that will heal with drainage. At the time of surgery, prevention would require either endoscopic control of the mucosal remnants, in case of sinus exclusion, or leaving the sphenoid sinus air-filled under a sealed sella. PMID- 9749547 TI - Thirty-five-millimeter photography using the Kantor-Berci video laryngoscope. AB - The Kantor-Berci model II laryngoscopes employ a centrally located rigid telescope. Although the primary application is for video laryngoscopy, the system can also be used for 35-mm photography during microlaryngeal operations. The fixed, unchangeable field of view and the great depth of focus make this system ideal for photographic documentation during endolaryngeal surgery without interruption of the procedure. PMID- 9749546 TI - Effects of angiogenic growth factors and a penetrance enhancer on composite grafts. AB - Skin-cartilage composite grafts are invaluable tissues used in facial reconstruction, yet their survival is unpredictable beyond a 1-cm diameter. In this study, the angiogenic growth factors basic fibroblast growth factor (bFGF) and endothelial cell growth factor (ECGF) and a penetrance enhancer (dimethyl sulfoxide [DMSO]) were applied to composite grafts to determine their effects on survival and vascularization. We applied ECGF, bFGF, and DMSO either topically or by intradermal injection to 120 auricular composite grafts (3.0 cm diameter) in New Zealand White rabbits. Dermabrasion was performed in 2 groups to attempt to increase transdermal delivery. Graft viability and vascularity were evaluated 3 weeks later by template analysis and angiography. In the results, ECGF and bFGF, when grouped together, had a 40% increase in vascular ingrowth as compared to controls (p < .001). However, neither ECGF nor bFGF increased graft survival. A coincidental finding was that DMSO with dermabrasion significantly improved graft viability (>100%) with or without an angiogenic agent (p < .02). The potential of DMSO with dermabrasion to increase composite graft viability warrants further investigation. PMID- 9749548 TI - Electroporation therapy of head and neck cancer. AB - The purpose of this paper is to introduce the concept of electroporation therapy and present our results from using this new technique combined with intralesional bleomycin in head and neck cancer patients. Electroporation therapy is a technique wherein high-voltage electric impulses delivered into a neoplasm transiently increase cell membrane permeability to large molecules, including cytotoxic agents. In this phase I/II study, extremely low-dose bleomycin sulfate was electroporated into head and neck malignant neoplasms in 10 patients. Tumor responses included 2 nonresponders, 3 partial responders, and 5 complete responders, with a mean follow-up of 40 weeks. We conclude that this technique offers promising possibilities in the local treatment of head and neck cancer. PMID- 9749549 TI - External rhinoplasty approach for extirpation and immediate reconstruction of congenital midline nasal dermoids. AB - Surgical extirpation is the treatment of choice for nasal dermoids in the pediatric population. Approaches include vertical-dorsal rhinotomy, lateral rhinotomy, transverse rhinotomy, and external rhinoplasty. While short-term results suggest that the last approach is cosmetically most acceptable, the required wide undermining of the nasal tip and dorsum may affect subsequent nasal growth. To assess long-term results, we reviewed our experience with 8 cases encountered over the past 10 years. Modifications of the technique to minimize midline dorsal depression are described. PMID- 9749550 TI - Tracheal neoplasms in children. AB - Primary tracheal neoplasms are extremely rare lesions in the pediatric age group. This study reviews the English-language literature to better characterize these lesions in children and reports 2 additional patients. Reports of only 36 infants and children through adolescence with primary tracheal neoplasms were discovered after an exhaustive literature review of the last 30 years. The data are analyzed with regard to pathology, demographics, symptomatology, site, and percent luminal obstruction. We report 2 additional patients with photographic documentation, imaging studies, and histopathology. Of the 36 previously reported lesions, 64% were characterized as benign and 36% as malignant. Fifty-six percent of all lesions were initially misdiagnosed as asthma. The most common site was the posterior membranous wall of the cervical trachea. In 14 (39%) of the 36 patients, the lesions obstructed more than 50% of the lumen at the time of diagnosis. The timely diagnosis of tracheal masses depends upon maintaining a high index of suspicion and conducting an efficient workup, including definitive evaluation by bronchoscopy. The evaluation and the differential diagnosis of tracheal neoplasms in the pediatric population is discussed. PMID- 9749551 TI - Pharyngeal fibrovascular polyp in a child. AB - Fibrovascular polyp of the upper aerodigestive tract is an uncommon tumor that may present in pediatric patients with symptoms ranging from dysphagia to asphyxiation and death. We present a unique case of a pediatric patient with an asymptomatic fibrovascular polyp noted as an incidental finding on a cervical ultrasound evaluation. This lesion extended from the posterior tonsillar pillar and prolapsed freely into the nasopharynx and esophagus. The literature relevant to this case is reviewed, and the etiology, pathophysiology, and management principles are discussed. PMID- 9749552 TI - Clinical and pathologic characterization of mucosa-associated lymphoid tissue lymphoma of the head and neck. AB - Mucosa-associated lymphoid tissue (MALT) has recently been recognized as a possible site of origin for low-grade lymphomas of the B-cell type. Though relatively rare, these MALT lymphomas may arise within several sites in the head and neck, and often present diagnostic and therapeutic challenges. We review 4 cases of primary MALT lymphoma of the head and neck, treated with surgical excision (3 cases), irradiation (2 cases), and chemotherapy (1 case), to further characterize this new subtype of head and neck malignancy. The mean time from onset of symptoms to histologic diagnosis was 15 months. Fine needle aspiration identified an atypical lymphoid infiltrate in only 1 of 3 patients. Immunohistochemical analysis was essential in establishing the diagnosis of MALT lymphoma in all 4 of the cases, and demonstrated characteristic negative staining for CD3, CD5, and CD43, positive staining for CD20, and monotypic staining for either kappa or lambda light chain immunoglobulin markers. All patients achieved complete remission after primary therapy, and all remain free of disease with follow-up ranging from 6 to 54 months (mean 33 months). The diagnosis of MALT lymphoma should be considered in cases of atypical lymphoid infiltrates in the head and neck, and increased awareness coupled with detailed immunohistochemical analysis is essential to securing an accurate diagnosis. Clinical remission of MALT lymphoma may be achieved with several modalities, but further study will be required to determine the long-term response to treatment. PMID- 9749553 TI - Computed tomographic findings in two cases of cellulitis of the infratemporal fossa with abscess formation. PMID- 9749554 TI - Alveolar soft part sarcoma of the head and neck region. AB - Alveolar soft part sarcoma is a soft tissue malignancy most often found in the extremities of young adults; when these tumors arise in the head and neck area, they usually appear in the orbit or the tongue. Their initial behavior is relatively indolent, but over time a sizable number of these tumors recur locally and metastasize; as such, they are best regarded as fully malignant neoplasms. The derivation of these tumors remains uncertain: while some have suggested that these are tumors of muscle origin and others have postulated a neuroendocrine origin, the evidence accumulated to date is conflicting, and so these neoplasms continue to be regarded as tumors of uncertain origin. Surgical excision is the mainstay of therapy. PMID- 9749555 TI - Cellular deformation: mechanics and mechanisms of physiologic response: introduction to the Emory Symposium. PMID- 9749556 TI - Mechanical deformation of the arterial wall in hypertension: a mechanism for vascular pathology. AB - Hypertension is an independent risk factor for atherosclerotic disease. It has been proposed that the atherogenic potential of hypertension is due to the development of a proinflammatory state within the arterial wall that is, at least in part, a result of the generation of reactive oxygen species. This article proposes that mechanical deformation of the arterial wall is a critical stimulus in this scheme. Data is reviewed that demonstrate that mechanical deformation of the arterial wall stimulates the generation of reactive oxygen species and subsequently results in the upregulation of redox-sensitive proinflammatory gene products. PMID- 9749558 TI - The study of the influence of flow on vascular endothelial biology. AB - It is now recognized that the mechanical environment of a cell has an influence on its structure and function. For the vascular endothelial cell that resides at the interface of the flowing blood and the underlying vessel wall, there is mounting evidence of the importance of flow and the associated wall shear stress in the regulation of endothelial biology. Not only is it a sensitive regulator of endothelial structure and function, but different flow environments will influence endothelial cell biology differently. Furthermore, there may be an interaction between the chemical environment of a cell and its mechanical environment. This is illustrated by the inhibition by steady laminar shear stress of the cytokine induction of VCAM-1. Results also are presented in which flow studies have been conducted using a co-culture model of the vessel wall. These experiments provide evidence of a quiescent endothelium; however, much more needs to be done to engineer the cell culture environment to make it more physiologic. PMID- 9749557 TI - Loading paradigms--intentional and unintentional--for cell culture mechanostimulus. AB - Recent interest in the response of cells or tissues to mechanical stimuli has led to the introduction of a variety of laboratory devices designed to deliver quantified mechanical inputs to culture systems. Such devices commonly rely upon distention of a flexible culture substrate, achieved either by direct platen abutment or by transmural pressure differentials. Unfortunately, the substrate distentions in such systems are often unintentionally nonuniform, and typically also induce motions in the overlying liquid nutrient medium--motions which in turn exert unintended reactive stresses upon the culture layer. In order to characterize the nature of these reactive fluid stresses, computer models have been developed for the nutrient medium flow fields (ie, the velocity and pressure distributions) for three established contemporary cell culture mechanostimulus systems. Temporal and spatial distributions of reactive normal and shear stresses are reported for typical duty cycles in these respective instruments. PMID- 9749559 TI - Skeletal cell stresses and bone adaptation. AB - There is no tissue in which mechanical stresses have been studied in more detail than the skeletal system, this focus arising primarily because bone plays a clear structural role in the body. However, the hypothesis that the skeleton represents an optimally designed structure has contributed remarkably little to our understanding of the development and adaptive capabilities of bone tissue. Recent investigations on the consequences of mechanical, hydrostatic, and electrical stresses on the cells of bone tissue have served to redirect the discussion of bone modeling and remodeling processes. These studies have refocused attention on the importance of chronic low-level dynamic stresses in mediating the physiologic response of bone tissue. Important recent observations suggest that an approach premised on the self-organizational properties of bone tissue may lead to significant improvements in our understanding and control of bone morphologic development, adaptation, and healing. PMID- 9749560 TI - Strain amplification in the bone mechanosensory system. AB - This article discusses the potential mechanisms by which the strain induced at the membrane of an osteocyte may be amplified from the strain experienced by the whole bone due to mechanical loading. These mechanisms address the question of how these mechanical load-induced small strains of (typically) about 0.1% (but up to 0.5%) applied to a whole bone are amplified to strains of 1% or larger at the membrane of the osteocyte buried in its lacuna in the bone matrix. The answer to this question is an important link in the mechanosensory system in bone and in relating in vitro cell studies to in vivo cellular response. PMID- 9749561 TI - Experimental elucidation of mechanical load-induced fluid flow and its potential role in bone metabolism and functional adaptation. AB - Several researchers have developed theories implicating some manifestation of mechanical forces such as stress, strain, and strain energy density for the initiation of cellular processes associated with functional adaptation. The mechanisms underlying dynamic bone growth and repair in response to mechanical stimuli, however, are not fully understood. Load-induced fluid flow has been postulated to provide a mechanism for the transmission of mechanical signals (eg, via shear stresses, enhancement of molecular transport, or electrical effects) and the subsequent elicitation of a functional adaptation response in bone. Although indirect evidence for such fluid flow phenomena can be found in the literature pertaining to strain-generated potentials, experimental studies are inherently difficult. This motivated the authors to develop theoretical as well as ex vivo, in vitro, and in vivo experimental methods for the study of transport processes and fluid flow within bone under well-controlled mechanical loading conditions. By introducing tracer substances such as disulphine blue, procion red, and microperoxidase into the experimental system, transport and fluid flow could be visualized at tissue, cellular, and subcellular levels, respectively. Based on these studies, it could be shown that load-induced fluid flow represents a powerful mechanism to enhance molecular transport within compact bone tissue. Furthermore, the distribution of transport-elucidating tracers is a function of mechanical loading parameters as well as the location within the cross-section of the bone cortex. PMID- 9749562 TI - Effect of mechanical strain on expression of Na+,K+-ATPase alpha subunits in rat aortic smooth muscle cells. AB - This article reviews related studies from the authors' laboratory, which focus on the regulation of vascular Na+,K+-ATPase in hypertension. Earlier studies, including the authors', suggested that Na-pump activity in cardiovascular tissues is subject to regulation during hypertension; most of these studies report a stimulation of the vascular enzyme during established stages of hypertension. To test hypothesis that in vascular smooth muscle, strain resulting from elevated pressure may be a signal initiating a cascade of events leading to increased expression of Na+,K+-ATPase, the authors used cell culture and the Flexercell Strain Unit to apply cyclical stretch to rat aortic smooth muscle cells (ASMC) for several days. These studies demonstrated that mechanical strain induces the upregulation of both the alpha-1 and alpha-2 subunits of Na+,K+-ATPase. Mechanisms underlying these changes appear to involve a transient increase in intracellular sodium entering the cell through stretch-activated channels. Calcium entering the cell via L-type channels did not affect stretch-induced upregulation of the alpha isoforms. In addition, protein kinase C inhibition resulted in inhibition of the Na-pump during stretch, but not under nonstretch conditions. The authors conclude that the stretch component of vascular pressure upregulates the Na+,K+-ATPase catalytic subunits. Intracellular sodium may be a signal for this regulation. In addition, phosphorylation by PKC may be important in stretch-induced short-term regulation of the vascular Na-pump. PMID- 9749563 TI - Effects of mechanical forces on lung-specific gene expression. AB - Fetal breathing movements (FBM) are necessary for fetal lung growth and maturation. The authors analyzed fetal rat lungs cultured with or without lung distension and tracheal ligation, and examined the effects of mechanical stretch on a human pulmonary epithelial cell line (NCI-H441) that shows regulated expression of surfactant proteins (SP-A, SP-B). Cells were grown on silastic membranes and mounted in a Flexercell Strain Unit. Cyclic deformation simulating FBM was achieved by applying a vacuum of 22 kPa (5%-15% radial deformation) at 50 cycles per minute for 2 to 24 hours. Results indicate that static distension for as little as 4 hours decreased steady-state SP-A and SP-B mRNA levels in whole lung (n = 5-6, P < .01). In contrast, cyclic stretching of H441 cells for 24 hours increased SP-B and SP-A expression 2- to 4-fold over controls. Cyclic deformation also significantly enhanced 3H-choline incorporation into saturated phosphatidylcholine. Dynamic mechanodeformation may be a critical stimulus for fetal lung development. PMID- 9749564 TI - Paracrine mediators of mechanotransduction in lung development. AB - The process of normal fetal lung development is dependent on "mild" tissue distension (approximately 3 mm Hg) by fluid, resulting in the production of pulmonary surfactant which is necessary for survival at the time of birth. The mechanical "stretching" of lung tissue triggers a cellular differentiation cycle, in part by stimulating the expression and production of cell phenotype-specific soluble cytokines. Pulmonary cytokines regulate differentiation and metabolic function of neighboring cells. For example, tonic stretching of type II alveolar epithelial cells in monolayer culture stimulates the expression and production of the differentiation factor parathyroid hormone-related peptide (PTHrP), which is released by type II cells and specifically binds to its receptor on contiguous fibroblasts, stimulating the "second messenger" cyclic AMP. Tonic distension of cultured type II cells increases PTHrP production, and distension of fibroblasts in monolayer culture increases their PTHrP responsiveness, suggesting that stretching couples and coordinates the production and receptor-mediated action of PTHrP. These data provide a mechanistic basis for the previously observed hand-in glove spatial pattern of PTHrP and the PTHrP receptor (PTHrPR) in developing terminal airways. PTHrP stimulates specific differentiated functions of fetal lung fibroblasts by: 1) augmenting glucocorticoid binding; 2) increasing metabolic activities directly related to surfactant synthesis, such as lipoprotein lipase elaboration and triglyceride uptake rate; 3) stimulating cytokines, such as interleukins 6 and 11, that can act in a retrograde fashion on epithelial cells; 4) thereby increasing the synthesis of surfactant phospholipids and surfactant-associated proteins, closing this stretch-mediated cell-cell interactive loop. Experimental interruption of this mechanism at any of these steps blocks the spontaneous maturation of the lung in vitro, as evidenced by the inhibition of surfactant production. PMID- 9749565 TI - Use of echocardiography in patients with known or suspected infective endocarditis. PMID- 9749566 TI - Changes in peripheral blood levels and pulse frequencies of GnRH in patients with hypopituitarism. AB - Pituitary dysfunction occasionally results from brain tumors or the surgical resection of brain tumors. The authors examined two patients with hypogonadotropic secondary amenorrhea, who had undergone surgical removal of brain tumors. Changes in immunoreactive gonadotropin-releasing hormone (GnRH) secretion are of interest in patients with a gonadotropin and gonadal steroid deficit, because both steroid and pituitary feedback systems are altered by tumors or tumor resection. The authors thus measured GnRH, luteinizing hormone, and follicle-stimulating hormone levels every 15 minutes for 4 hours by radioimmunoassay and investigated qualitative and quantitative changes in the pulsatile patterns of these hormones in two hypogonadotropic hypogonadism patients. They also performed similar multiple measurements of GnRH in two normal cycle women in follicular phase and two postmenopausal women. The concentration of plasma GnRH in two hypopituitarism patients was compared with that in two normal cycle women and two postmenopausal women. The study showed that the peripheral blood level of GnRH was significantly lower in two hypopituitarism patients than in both normal cycle and postmenopausal women, and that the pulsatile frequency was not different among these three groups. These findings suggest that alteration of feedback systems results in a decrease in the blood level of GnRH, and that pulses of GnRH maintain normal fluctuation despite the alteration of the hormonal circumstances in two hypogonadotropic hypogonadism patients. PMID- 9749567 TI - Emission spectra of colonic tissue and endogenous fluorophores. AB - Autofluorescence emission spectra of normal, adenomatous, and malignant tissues of the colon were compared to that of known fluorophores to indicate the possible causes of tissue fluorescence. Data were collected from normal mucosa (n = 18), adenomatous polyps (n = 32), and adenocarcinoma (n = 18) of the colon. A range of cellular and extracellular fluorophores (elastin, collagen, flavin adenine dinucleotide, nicotinamide adenine dinucleotide, phenylalanine, pyridoxal 5' phosphate, tryptophan, and tyrosine) were similarly examined using a spectrofluorometer with emission and excitation spectrometers. Emission intensities were plotted against wavelength. Wavelengths of peak emission and the width of each peak at half its maximum intensity were measured. Colonic tissue gave four major emission peaks, the wavelengths of which were independent of tissue histology. Tryptophan and collagen type IV appeared to be responsible for two of the peaks. It is possible that NADH may be the cause of a third emission maxima. PMID- 9749568 TI - The natural history of Parkinson's disease. AB - There are still insufficient data on the natural course of Parkinson's disease (PD) owing to lack of standardized longitudinal follow-up studies. Reported progression rates in early PD vary considerably by a factor of 2 to 3. Similarly, data from sequential [18F]dopa PET studies in PD patients have produced variable decline rates of PET indices ranging between 7 and 70% per decade. Risk factors for rapid progression include old age at onset, concomitant major depression, dementia, and akinetic-rigid symptom presentation. The introduction of levodopa into the routine treatment of PD patients had a dramatic impact on symptomatic control without affecting the underlying rate of disease progression. By contrast, monoamine oxidase (MAO) B inhibition by deprenyl monotherapy in early PD was shown to delay the need for levodopa by around 9 months. However, the neuroprotective action disappeared after 2 years of follow-up. Furthermore, deprenyl also failed to influence the subsequent development of levodopa-induced motor complications. Available studies on mortality in PD provide heterogeneous mortality rates, probably because of discrepancies between patient populations with respect to co-morbidity, disease stage at study entry, and diagnostic accuracy. However, the most recent follow-up from the DATATOP cohort suggests normal life expectancy in carefully selected patients without significant co morbidity and with adequate treatment and expert follow-up. PMID- 9749569 TI - The early diagnosis of Parkinson's disease. AB - Current accepted clinical criteria for the diagnosis of Parkinson's disease (PD) provide high sensitivity for detecting parkinsonism but generally show poor specificity for identifying brainstem Lewy body disease. Biochemical markers that can be used to reliably diagnose clinical and preclinical PD have thus far been sought unsuccessfully. It is now known that some PD kindreds have a mutation of the alpha-synuclein gene, but this cannot be used as a genetic marker for most familial and sporadic cases. Functional imaging provides a means of discriminating typical from atypical PD, revealing characteristic patterns of loss of dopaminergic function. In addition, PET and SPECT show preserved levels of striatal metabolism and dopamine receptor binding in PD, whereas levels are reduced in the atypical variants. [18F]Dopa PET can also detect preclinical PD. In one series there was a reported 40% prevalence of preclinical dopaminergic dysfunction in asymptomatic adult relatives of familial PD patients. Finally, PET and SPECT can both be used to follow PD progression objectively. Such studies suggest an annual 4 to 12% loss of dopamine terminal function in early PD and a preclinical disease window of only a few years. In the future, functional imaging is likely to play an increasingly important role in assessing the efficacy of putative neuroprotective agents. PMID- 9749571 TI - Determinants of neuronal vulnerability in neurodegenerative diseases. AB - Selective neuronal vulnerability can be defined anatomically by the differential vulnerability of circuits and neurochemically by the vulnerability of neurons that differentially express particular proteins. The anatomic perspective is exemplified by the vulnerability of the nigrostriatal projection in Parkinson's disease (PD), the degeneration of upper and lower motor neurons in amyotrophic lateral sclerosis (ALS), and the preferential loss of long corticocortical projections in Alzheimer's disease (AD). The neurochemical perspective is reflected in the heightened vulnerability of neurons that normally express high somatodendritic levels of neurofilament (eg, entorhinal and association cortices in AD, the spinal cord in a mouse model of ALS, and the retina in a primate model of glaucoma), as well as the reduced vulnerability of neurons that express calcium-binding proteins (eg, neocortex of AD patients, the spinal cord and brainstem of ALS patients, and the spinal cord of a mouse model of ALS). By combining neurochemical and anatomic correlates of vulnerability, an integrated view of vulnerable neurons is emerging in which characteristics of vulnerable neurons appear to transcend both brain region and disease state, suggesting that neurodegenerative disorders share common mechanisms of degeneration. PMID- 9749570 TI - Alzheimer's disease and Parkinson's disease: distinct entities or extremes of a spectrum of neurodegeneration? AB - Alzheimer's disease (AD) and Parkinson's disease (PD) are generally considered to be separate and distinct disease entities. However, a considerable amount of evidence demonstrates that these disorders share common clinical and neuropathologic features and that overlap between the two conditions is extensive. For example, a significant percentage of AD patients exhibit extrapyramidal features, and many PD patients develop dementia. Similarly, at autopsy many AD patients not only exhibit the neuropathologic features of that disorder but also exhibit nigral pathology, including Lewy bodies. The vast majority of demented PD patients show widespread neurofibrillary tangles and senile plaques as well as Lewy body formation and nigral degeneration. The extent of such overlap is far greater than one would anticipate by chance alone. We argue that such overlap reflects a common pathogenic mechanism for the neurodegeneration encountered within specific vulnerable neuronal populations. Furthermore, we suggest that the current nosologic approach, which attempts to separate AD from PD, fails to properly deal with the issue of overlap and that a new classification of the neurodegenerative disorders should be considered. PMID- 9749572 TI - Epidemiology versus genetics in Parkinson's disease: progress in resolving an age old debate. AB - Determining the relative contributions of environment and heredity to the cause of Parkinson's disease (PD) is more than an academic issue because its resolution dictates future research directions to an enormous degree. This article reviews new advances on both sides of this equation. The recent identification of the genetic mutation responsible for parkinsonism in a large Italian kindred is likely to provide exciting new research opportunities but the mutation does not appear to be responsible for the vast majority of PD. A large twin study also points away from genetic influences as important, at least in patients with disease beginning after the age of 50 years. On the other hand, genetic influences loom large in younger-onset disease. With regard to the environment, epidemiologic studies have provided only broad, thought-tantalizing clues to the cause of the disease. Although rural living, well-water consumption, and exposure to pesticides have emerged as potential risk factors, identification of specific agents is lacking, and aging remains as the only unequivocal risk factor for the disease. The surprisingly strong inverse relationship between cigarette smoking and PD provides an intriguing lead, but novel experimental avenues to pursue this observation are not readily obvious. The amyotrophic lateral sclerosis/dementia/parkinsonism complex in the western Pacific suggests the possibility of long-latency toxins, but pinning down a specific causative agent for this syndrome has eluded investigators to date. Despite the many obstacles ahead, however, research on PD appears to be more robust than ever, and our quest to find its cause appears to be under a full head of steam as we approach the 21st century. PMID- 9749573 TI - Genetics of Parkinson's disease. AB - A genetic contribution to the etiology of Lewy body Parkinson's disease (PD) is now well established, based on the demonstration of a familial aggregation of the disease by case-control and twin studies and on the description of large multigenerational families in whom clinically and pathologically typical PD is inherited in an autosomal dominant fashion. In the largest of these families, a gene locus has been mapped to the long arm of chromosome 4 and a putative disease causing mutation has been identified in the gene for alpha-synuclein. However, analysis of a large number of individuals with sporadic and familial PD suggests that a mutation in this gene is a very rare cause of the disorder, either familial or sporadic. Another locus for autosomal dominantly inherited Lewy body PD has recently been mapped to chromosome 2p13. Possible strategies for the identification of further PD genes are discussed. PMID- 9749574 TI - Genetic risk factors in Parkinson's disease. AB - There is now overwhelming evidence for a role of genetic factors in susceptibility to Parkinson's disease. This review concentrates only on the evidence from candidate association studies because the role of linkage analysis and the successes of positional cloning are dealt with by others in this supplement. Candidate gene studies are a useful and powerful adjunct to the armamentarium of the researcher, but the robustness of the data is difficult to achieve. This is illustrated by the large literature on this subject, only a relatively small proportion of which has been reproduced. A recent study by our group is used to illustrate some of these difficulties, and the possible ways around some of the inherent biases in most of the studies are also discussed. PMID- 9749575 TI - Autosomal dominant Parkinson's disease and alpha-synuclein. AB - Multiple factors have been hypothesized over the years to be contributory and/or causative for Parkinson's disease (PD). Hereditary factors, although originally discounted, have recently emerged in the focus of PD research. The study of a large Italian family with PD using a genome scan approach led to the mapping of a PD susceptibility gene to the 4q21-q23 genomic region, where the gene for alpha synuclein was previously mapped. Mutation analysis of the alpha-synuclein in four unrelated families with PD revealed a missense mutation segregating with the illness. Alpha-synuclein is an abundant presynaptic protein in the human brain with unknown function. It is conceivable that the mutation identified in the PD families may result in self-aggregation and/or decreased degradation of the protein, leading to the development of intracytoplasmic inclusion bodies and eventually to neuronal cell death. Moreover, the discovery of a mutation in the synuclein gene may offer us new insights in the understanding of the pathways that lead to neuronal degeneration. PMID- 9749576 TI - Structure/function in neuroprotection and apoptosis. AB - The three-dimensional conformation of proteins influences their potential to function correctly within the cell. Protein conformational issues are particularly important in neurodegeneration, as has been shown by misfolded protein forming the basis of plaques in Alzheimer's disease and prion diseases. This article focuses on protein structure/function specifically for proteins important in the pathogenesis of neurodegenerative conditions and those involved in apoptosis. The proteins used as examples in this review include alpha synuclein, the promyelocytic leukemia protein, and glyceraldehyde 3-phosphate dehydrogenase. PMID- 9749577 TI - Understanding cell death in Parkinson's disease. AB - Current concepts of the cause of Parkinson's disease (PD) suggest a role for both genetic and environmental influences. Common to a variety of potential causes of nigral cell degeneration in PD is the involvement of oxidative stress. Postmortem analysis shows increased levels of iron, decreased complex I activity, and a decrease in reduced glutathione (GSH) levels. The decrease in GSH levels may be a particularly important component of the cascade of events leading to cell death because it occurs in the presymptomatic stage of PD and may directly induce nigral cell degeneration or render neurons susceptible to the actions of toxins. There is evidence suggesting that oxidative stress might originate in glial cells rather than in neurons, and alterations in glial function may be an important contributor to the pathologic process that occurs in PD. Oxidative damage occurs in the brain in PD, as shown by increased lipid peroxidation and DNA damage in the substantia nigra. Increased protein oxidation is also apparent, but this occurs in many areas of the brain and raises the specter of a more widespread pathologic process occurring in PD to which the substantia nigra is particularly vulnerable. The inability of the substantia nigra to handle damaged or mutant (eg, alpha-synuclein) proteins may lead to their aggregation and deposition and to the formation of Lewy bodies. Indeed, Lewy bodies stain for both alpha synuclein and nitrated proteins. Current evidence enables us to hypothesize that a failure to process structurally modified proteins in regions of the brain exhibiting oxidative stress is a cause of both familial and sporadic PD. PMID- 9749578 TI - Advanced glycation end products in neurodegeneration: more than early markers of oxidative stress? AB - Oxidative stress is believed to play a decisive role in the pathogenesis of Parkinson's disease (PD). In addition, Lewy bodies, densely crosslinked intracellular protein deposits formed from cytoskeletal components, accumulate in presymptomatic stages of the disease. Recent findings indicate that "advanced glycation end products" (AGEs) are the major structural crosslinkers that cause the transformation of soluble neurofilament proteins to insoluble Lewy bodies. AGE formation is increased under conditions of oxidative stress, such as early GSH depletion, that are evident in the substantia nigra of PD patients, and is inhibited by radical scavengers and thiol antioxidants. Because AGEs not only are markers of oxidative stress but are also active participants in cell signaling by activation of glial cells to produce superoxide and nitric oxide, they can be considered part of a vicious cycle, which finally leads to neuronal cell death in the substantia nigra in PD. PMID- 9749579 TI - Mitochondria in the etiology and pathogenesis of Parkinson's disease. AB - Mitochondria play a critical role in cellular energy metabolism. The identification of a respiratory chain defect in Parkinson's disease (PD) provides not only a direct link with toxin models of parkinsonism but also insight into the mechanisms involved in etiology and pathogenesis. The presence of the complex I deficiency in PD substantia nigra and platelets suggests the involvement of a systemic cause. Genomic transplantation studies have been undertaken that involve the transfer to a novel nuclear background of mitochondrial DNA (mtDNA) from PD patients with a complex I defect, followed by both mixed and clonal expansion of the resulting cybrids. The mixed cybrids with the PD mtDNA expressed the complex I defect present in the original PD donor platelets. Clonal expansion of one such mixed cybrid culture produced a spectrum of clones with complex I and complex IV activities, ranging from severe deficiency to normal range, a pattern typical of a heteroplasmic mtDNA mutation. Histochemical, immunohistochemical, and functional assessments of delta psi(m) all showed a pattern in the PD clones typical of that produced by a mtDNA mutation. Patients with focal dystonia and a platelet complex I defect were used as disease controls for the cybrid studies. The mitochondrial abnormality was eradicated by transfer of dystonia mtDNA to a control nuclear background in both mixed and clonal cybrids, with no evidence of clonal heterogeneity. These results help to validate our findings in the PD patients and suggest that the complex I deficiency in dystonia is not due to an abnormality of mtDNA. We hypothesize that the mtDNA defect alone may be the cause of PD in a proportion of patients and may contribute to pathogenesis in others. Identification of the mtDNA genotype responsible for PD may allow the testing of neuroprotective strategies in appropriate patients. PMID- 9749580 TI - Mitochondrial dysfunction in Parkinson's disease. AB - This review discusses the etiology and pathogenesis of Parkinson's disease (PD). Mitochondrial respiratory failure and oxidative stress appear to be two major contributors to nigral neuronal death in PD. Complex I deficiency has been reported by several groups and appears to be one of the basic abnormalities responsible for mitochondrial failure. The principal question is whether or not complex I deficiency is primary or secondary. The second question is whether or not complex I deficiency is localized in the nigrostriatal system or is systemically present. It is our impression that complex I deficiency is not the primary cause but that its deficiency appears to be systemic. The primary cause may be the combination of genetic background and potential nigral neurotoxins. Exposure of nigral neurons to a high risk for oxidative damage because of its high dopamine content may be the reason for more pronounced nigral complex I deficiency compared to systemic organs. Oxidative stress and mitochondrial failure produce a vicious cycle in nigral neurons. To explore the genetic risk factors of sporadic PD, studies on familial PD and parkinsonism are important. Recently, an autosomal dominant form of familial PD was found to be caused by point mutations of the alpha-synuclein gene, and an autosomal recessive familial parkinsonism was mapped to the long arm of chromosome 6 near the Mn-SOD gene locus. Information obtained in these familial cases will contribute to the research on sporadic PD. PMID- 9749582 TI - Glial cells and inflammation in Parkinson's disease: a role in neurodegeneration? AB - The data reviewed here show that, in Parkinson's disease (PD), some dopaminergic neurons are more vulnerable than others to the pathologic process. The glial cells surrounding dopaminergic neurons may be involved in this selective vulnerability. One subpopulation of glial cells, in particular, may play a neuroprotective role by metabolizing dopamine and scavenging oxygen free radicals that are associated with dopamine metabolism. Another subpopulation of glial cells may be deleterious to dopaminergic neurons. This effect may be mediated by the production of nitric oxide and cytokines, which may in turn account for the oxidative stress observed in the substantia nigra of patients with PD. Finally, this inflammatory reaction may result in the induction of apoptosis. PMID- 9749581 TI - Excitotoxicity and nitric oxide in Parkinson's disease pathogenesis. AB - A potential role for excitotoxic processes in Parkinson's disease (PD) has been strengthened by the recent observations that there appears to be a mitochondrially encoded defect in complex I activity of the electron transport chain. An impairment of oxidative phosphorylation will enhance vulnerability to excitotoxicity. Substantia nigra neurons possess N-methyl-D-aspartate receptors and there are glutamatergic inputs into the substantia nigra from both the cerebral cortex and the subthalamic nucleus. After activation of excitatory amino acid receptors, there is an influx of calcium followed by activation of neuronal nitric oxide (NO) synthase, which can then lead to the generation of peroxynitrite. Consistent with such a mechanism, studies of 1-methyl-4-phenyl 1,2,3,6-tetrahydropyridine neurotoxicity in both mice and primates have shown that inhibition of neuronal NO synthase exerts neuroprotective effects. Studies utilizing excitatory amino acid receptor antagonists have been inconsistent in mice but show significant neuroprotective effects in primates. These results raise the prospect that excitatory amino acid antagonists for neuronal NO synthase inhibitors might be useful in the treatment of PD. PMID- 9749583 TI - Neuroprotective and neurorestorative properties of GDNF. AB - Glial cell line-derived neurotrophic factor (GDNF) promotes recovery of the injured nigrostriatal dopamine system and improves motor functions in both rodent and nonhuman primate models of Parkinson's disease (PD). The neurorestorative effects of a single administration of GDNF last for at least 1 month and can be maintained in rhesus monkeys by monthly injections. Adult midbrain dopamine neurons stimulated by GDNF show increased cell size, neurite extent, and expression of phenotypic markers. In parkinsonian nonhuman primates, GDNF treatment improves three of the cardinal features of PD: bradykinesia, rigidity, and postural instability. Although intracerebral administration is necessary because of the blood-brain barrier, intraventricular, intrastriatal, and intranigral routes of administration have been found to be efficacious in rodents and nonhuman primates. GDNF also induces neuroprotective changes in dopamine neurons which are active within hours after trophic factor administration. The powerful neuroprotective and neurorestorative properties of GDNF seen in preclinical studies suggest that trophic factors may play an important role in treating PD. PMID- 9749584 TI - Programmed cell death: does it play a role in Parkinson's disease? AB - In recent years, the possibility that programmed cell death (PCD), which is mediated by genetic programs intrinsic to the cell, may underlie the degeneration of neurons that occurs in Parkinson's disease (PD) and allied disorders has become an important hypothesis. Although PCD was originally identified in tissues as a normal developmental phenomenon, there is no question that it can also occur in neurologic disease and models thereof. The possibility that PCD could occur in dopamine neurons in degenerative disease is made plausible by the observations that natural cell death, with the morphology of apoptosis, does occur in these neurons and that this event is regulated by developmental target interactions. In addition, it has been shown that apoptotic death can be induced in these neurons in some animal models of parkinsonism. We have shown, for example, that apoptosis can be induced during development by intrastriatal injection of the neurotoxin 6 hydroxydopamine. Other investigators have shown that apoptosis can be induced in a chronic model of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyride toxicity. However, investigations in human PD brains have yielded mixed results thus far, with some investigators identifying evidence of apoptotic death but others not. Further investigation of human postmortem tissue will benefit from a more complete understanding of the molecular basis of PCD in dopamine neurons, such that its molecular features can be investigated, rather than strictly relying on the morphologic markers presently available. PMID- 9749585 TI - Mitochondria in neurodegenerative apoptosis: an opportunity for therapy? AB - Apoptotic cell death has been shown to constitute the terminal process in some neurodegenerative diseases, notably Alzheimer's disease and Parkinson's disease (PD). A decrease in mitochondrial membrane potential (delta psiM) causing opening of the permeability transition pore (PTP) in mitochondrial membranes has been implicated as a critical effector of apoptosis in a variety of non-neural cells. Opening of the PTP leads to the release of so-called apoptosis initiation factors that induce the degradative events of apoptosis, such as nuclear chromatin condensation and DNA fragmentation. We have extended those findings to a neuronal model of apoptosis caused by trophic withdrawal, by showing that a decrease in delta psiM is an early event occurring 2 to 6 hours before the degradative events of apoptosis. A deficiency in mitochondrial complex I activity has been demonstrated in the substantia nigra of postmortem brains and several peripheral tissues obtained from PD patients. Because delta psiM is generated by the pumping of protons out across the inner mitochondrial membrane at the mitochondrial complexes, particularly complex I, we hypothesized that the decrease in complex activity could result in a decrease in delta psiM that would render PD substantia nigra neurons vulnerable to apoptosis. In preliminary studies, we have found a decrease in delta psiM in fibroblasts obtained from some PD patients. If a decrease in delta psiM consequent on decreased complex activity is an intrinsic defect in some PD patients, it would open a number of new avenues for the reduction of neuronal apoptosis in PD. The oncoprotein BCL-2 and the scavenger protein SOD-1 have been shown to reduce apoptosis by facilitating closure of the PTP. A number of agents have been shown to maintain BCL-2 and/or SOD-1 synthesis in damaged nerve cells and thereby reduce apoptosis. Other agents, such as cyclosporin A and some benzodiazepine receptor-binding agents, have been found to act directly on the PTP to reduce apoptosis. Accordingly, agents that maintain delta psiM and PTP closure may offer new and effective means of treating neurodegenerative apoptosis. PMID- 9749586 TI - A fluorescent double-labeling method to detect and confirm apoptotic nuclei in Parkinson's disease. AB - In situ end-labeling (ISEL) has become a widely used method to determine whether cells die via apoptosis by detecting double-stranded DNA breaks that are the result of endonuclease digestion. The enzyme terminal deoxynucleotidyl transferase can be used to label the digested 3'-OH ends of DNA with biotin-, digoxigenin-, or fluorescent probe-conjugated nucleotides. However, both single stranded and double-stranded DNA breaks can be labeled by this method and therefore ISEL cannot unequivocally demonstrate apoptosis when used alone. We have developed a fluorescent double-labeling method using ISEL combined with the cyanine dye YOYO-1 that binds to DNA. When combined with confocal laser microscopy and deconvolution analysis, YOYO-1 can demonstrate the presence or absence of nuclear chromatin condensation and thus confirm that ISEL-positive nuclei are indeed apoptotic. Preliminary findings indicate that dopaminergic neurons in the substantia nigra compacta die via apoptosis in Parkinson's disease. PMID- 9749587 TI - Levodopa toxicity and apoptosis. AB - Many in vitro studies have shown that levodopa is a potent toxin which is lethal to various cultured neuronal and non-neuronal cells. The in vitro toxicity of levodopa is linked mainly to its auto-oxidation, which generates a variety of harmful free radical species including superoxide, hydrogen peroxide, and hydroxyl radicals, and also semiquinones and quinones produced via the dopa melanin metabolic route. Such toxic effects of levodopa can be blocked by co treatment with antioxidants, particularly thiol-containing compounds. Several studies have shown that levodopa kills cells by triggering apoptosis, an active, intrinsic cell suicide program. Exposure of cultured neurons to levodopa induced the characteristic apoptotic cascade, including cell shrinkage, membrane blebbing, and nuclear and DNA fragmentation. Although levodopa is extremely toxic in vitro, there is no evidence that it damages nigrostriatal dopaminergic neurons in vivo in experimental animals and in patients with Parkinson's disease (PD). Likewise, although there is some evidence for the occurrence of apoptosis in the parkinsonian substantia nigra, it is not known whether levodopa administration is capable of inducing or accelerating programmed cell death of residual pigmented nigral neurons in PD. PMID- 9749588 TI - Neuroprotection for Parkinson's disease. AB - Levodopa is the most effective drug available for the treatment of Parkinson's disease (PD). As a precursor of dopamine, levodopa causes an increase in brain dopamine, which is depleted in PD. However, long-term levodopa therapy is associated with complications such as dyskinesias, motor fluctuations, and mental status changes. The treatment of these side effects is a major challenge for the clinician. PMID- 9749589 TI - DATATOP: a decade of neuroprotective inquiry. Parkinson Study Group. Deprenyl And Tocopherol Antioxidative Therapy Of Parkinsonism. AB - In 1987, the DATATOP clinical trial was initiated to examine the benefits of deprenyl (selegiline) and alpha-tocopherol in slowing the progression of Parkinson's disease (PD). After 14 +/- 6 (mean +/- SD) months of controlled observation, deprenyl 10 mg/day was found to significantly delay the time until enough disability developed to warrant the initiation of levodopa therapy. This effect was largely sustained during the overall 8.2 years of observation, including open-label deprenyl treatment and a second treatment randomization to continue deprenyl or switch to placebo. There were no accompanying benefits of deprenyl in postponing levodopa-related adverse effects or extending life. Alpha tocopherol produced no benefits. The 2.1% per year mortality rate of the DATATOP cohort was remarkably low, about the same as an age-matched population without PD. Neuroprotective therapy remains an elusive goal for the experimental therapeutics of PD. Advances in understanding pathogenesis, a robust pipeline of rational treatments, and the advent of valid and reliable biologic markers hold promise in the coming decade for developing and achieving neuroprotective therapies for PD. PMID- 9749590 TI - Dopamine agonists and neuroprotection in Parkinson's disease. AB - There is increasing interest in the potential of dopamine agonists to provide a neuroprotective effect and to alter the natural course of levodopa-treated Parkinson's disease (PD). Theoretically, such a protective effect might derive from (a) a levodopa sparing effect, (b) stimulation dopamine autoreceptors resulting in decreased dopamine synthesis, release, and turnover, (c) direct anti oxidant effects, and (d) restoration of dopaminergic tone to the dopamine denervated brain so as to restore inhibition to the subthalamic nucleus and thereby diminish STN-mediated excitotoxicity. Preclinical studies have demonstrated that dopamine agonists reduce dopamine formation in comparison to levodopa, protect cultured dopaminergic neurons from a variety of toxins including levodopa, and protect dopaminergic neurons from toxins and age-related degeneration in some rodent models of parkinsonism. Based on these findings, several clinical trials have been initiated in patients with early PD to test the effect of dopamine agonists on clinical and neuroimaging markers of disease progression. PMID- 9749591 TI - Subthalamic nucleus-mediated excitotoxicity in Parkinson's disease: a target for neuroprotection. AB - Dopamine deficiency causes disinhibition and overactivity of the subthalamic nucleus (STN). Output neurons from the STN are excitatory and use glutamate as a neurotransmitter. They project to the external and internal segments of the globus pallidum (GPe and GPi), the substantia nigra pars reticulata (SNr), and the pedunculopontine nucleus (PPN). In addition, STN neurons provide excitatory innervation to dopaminergic (DA) neurons in the substantia nigra pars compacta (SNc) that contain glutamate receptors. Stimulation of the STN induces bursting activity in SNc dopaminergic neurons. This raises the possibility that the disinhibition of STN neurons that occurs as a result of a dopamine lesion might induce excitotoxic damage in target structures, including the SNc. In addition, the reduction in complex I activity found in the nigra in Parkinson's disease (PD) may cause mitochondrial dysfunction and make SNc dopaminergic neurons vulnerable to even physiologic concentrations of glutamate. We postulate that the dopamine loss that occurs in PD produces augmented STN activity which, in turn, causes further damage to vulnerable dopaminergic neurons, thereby creating a scenario for an increasing cycle of neuronal loss in the SNc. In addition, STN overactivity could, in theory, cause damage to the GPi, SNr, and PPN and thereby account for the development of parkinsonian features that do not respond to levodopa in patients with advanced disease. This hypothesis suggests that pharmacologic or surgical therapies that reduce STN neuronal overactivity or block glutamate receptors in the SNc and other target structures might be neuroprotective and might slow or halt the progression of neurodegeneration in PD. PMID- 9749592 TI - The causes of Parkinson's disease are being unraveled and rational neuroprotective therapy is close to reality. AB - There has been significant progress in our knowledge of the cause, the pathogenesis, and the nature of the mechanism of cell death in Parkinson's disease (PD). Mutations in single genes have now been shown to be able to cause PD but likely only account for a small number of cases. Alternatively, there is evidence that environmental factors play a large role in the majority of cases of sporadic PD. Most likely, genetic factors predispose patients to develop PD if combined with other gene mutations or environmental toxins. Interest has thus focused on factors that contribute to the pathogenesis of neurodegeneration and the mechanism of cell death in an attempt to design a neuroprotective therapy. Oxidant stress, mitochondrial dysfunction, excitotoxicity with excess nitric oxide formation, and glia and inflammatory processes are all thought to contribute to the cell death process and agents that interfere with these events may be neuroprotective. It is now generally held that the final culmination of these events is the induction of apoptosis in nigral dopaminergic neurons and this too offers opportunities for providing neuroprotection. A rational argument can be made for investigating a large number of different approaches or combination of approaches in the hope of developing a meaningful neuroprotective therapy, using clinically relevant indices and neuroimaging markers of nigral dopaminergic neurons. It is evident that conventional approaches to trials that utilize large numbers of patients in search of small incremental effects are costly and time consuming. As such, it will be virtually impossible to test all of the potentially valuable neuroprotective agents that are now at hand, let alone those that will likely soon emerge. We suggest that it may be more profitable to test a large number of agents in a small number of selected patients in search of a more robust neuroprotective effect. In this way, we will reduce the risk of missing a powerful neuroprotective treatment with a treatment that might not otherwise have been studied because of a lack of time, money, or patients. PMID- 9749593 TI - Penetrating observations of dystonia. PMID- 9749594 TI - Rethinking brain tumors in babies and more. PMID- 9749595 TI - Functional brain networks in DYT1 dystonia. AB - Early-onset idiopathic torsion dystonia (ITD) is an autosomal dominant hyperkinetic movement disorder with incomplete penetrance, associated with a 3 base-pair deletion in the DYT1 gene on chromosome 9q34. To determine the metabolic substrates of brain dysfunction in DYT1 dystonia, we scanned 7 nonmanifesting and 10 affected DYT1 carriers and 14 normal volunteers with [18F]fluorodeoxyglucose and positron emission tomography. We found that DYT1 dystonia is mediated by the expression of two independent regional metabolic covariance patterns. The first pattern, identified in an analysis of nonmanifesting gene carriers was designated movement free (MF). This abnormal pattern was characterized by increased metabolic activity in the lentiform nuclei, cerebellum, and supplementary motor areas. The MF pattern was present in DYT1 carriers with and without clinical manifestations and persisted in DYT1 dystonia patients in whom involuntary movements were suppressed by sleep. The second pattern, identified in an analysis of affected gene carriers with sustained contractions at rest, was designated movement related (MR). This pattern was characterized by increased metabolic activity in the midbrain, cerebellum, and thalamus. The expression of the MR pattern was increased in waking DYT1 patients with sustained dystonia, compared with DYT1 carriers who were unaffected or who had dystonia only on action, as well as normal controls. MR subject scores declined significantly with sleep in affected DYT1 patients but not in normal controls. These findings indicate the penetrance of the DYT1 gene is considerably greater than previously assumed. ITD is mediated through the interaction of functional brain networks relating separately to gene status and to abnormal movement. PMID- 9749596 TI - Second malignancies in young children with primary brain tumors following treatment with prolonged postoperative chemotherapy and delayed irradiation: a Pediatric Oncology Group study. AB - Between 1986 and 1990, the Pediatric Oncology Group conducted a study in which 198 children younger than 3 years of age with malignant brain tumors were treated with prolonged postoperative chemotherapy in an effort to delay irradiation and reduce long-term neurotoxicity. Children younger than 2 years of age received 24 months of chemotherapy followed by irradiation, and those between 2 and 3 years of age received 12 months of chemotherapy plus irradiation. Chemotherapy was given in 28-day cycles (AAB, AAB), with cycle A = vincristine (0.065 mg/kg) intravenously on days 1 and 8 and cyclophosphamide (65 mg/kg) intravenously on day 1, and cycle B = cisplatinum (4 mg/kg) intravenously on day 1 and etoposide (6.5 mg/kg) intravenously on days 3 and 4. Five of the 198 children developed second malignancies, with a cumulative risk at 8 years of 11.3% (95% confidence interval [CI], 0-39%). Four of the five second malignancies occurred in children younger than 2 years of age at diagnosis, with a cumulative risk at 8 years of 18.9% (CI, 0-70%). Initial diagnoses were choroid plexus carcinoma (2 children), ependymoma (1 child), desmoplastic infantile ganglioglioma (2 children), and medulloblastoma (1 child). Duration from diagnosis of initial tumor to second malignancy was 33, 35, 57, 66, and 92 months. Three children younger than 2 years of age developed lymphoproliferative disease, that is, myelodysplastic syndrome (2 children), both with monosomy 7 deletions, and acute myelogenous leukemia (1 child), after 24 to 26 cycles of chemotherapy, including 8 cycles of etoposide. Two of 3 received craniospinal irradiation (2,560/3,840 cGy) and (3,520/5,320 cGy). Time to second malignancy was 7 years 8 months, 4 years 9 months, and 2 years 9 months. Two children developed solid tumors, at 5 years 6 months and 2 years 11 months, respectively, after initiation of treatment. A sarcoma developed after 26 cycles of chemotherapy and no irradiation, and a meningioma developed after 12 cycles of chemotherapy and local craniospinal irradiation. Potential causative factors for this high rate of secondary malignancies include prolonged use of alkylating agents and etoposide with or without irradiation. PMID- 9749597 TI - Time course of corticospinal excitability in reaction time and self-paced movements. AB - We used transcranial magnetic stimulation (TMS) to study the time course of corticospinal excitability before and after brisk thumb abduction movements, either in a simple reaction time (RT) paradigm or self-paced. Premovement increase in corticospinal excitability began about 20 msec earlier for self-paced compared with simple RT movements. For both simple RT and self-paced movements after electromyographic (EMG) offset, there was a first period of increased excitability from 0 to 100 msec, followed by a second period from 100 to 160 msec. Corticospinal excitability was decreased from about 500 to 1,000 msec after EMG offset for both types of movements. Our results show that motor preparation that begins 1.5 to 2 seconds before self-paced movement is not associated with increased corticospinal excitability. The first phase of increased corticospinal excitability after EMG offset may be due to activity of motor cortex neuron subthreshold for activating spinal motor neurons, and the second phase may reflect a subthreshold second agonist burst. The period of decreased corticospinal excitability after movement corresponds to the onset of event related synchronization (ERS) of electroencephalographic signals in the 20-Hz band, and supports the hypothesis that ERS may be related to an inactive, idling state of the motor cortex. PMID- 9749598 TI - Decreased striatal monoaminergic terminals in severe chronic alcoholism demonstrated with (+)[11C]dihydrotetrabenazine and positron emission tomography. AB - We used (+)[11C]dihydrotetrabenazine, a new ligand for the type 2 vesicular monoamine transporter, with positron emission tomography to study striatal monoaminergic presynaptic terminals in 7 male severe chronic alcoholic subjects without Wernicke-Korsakoff disease compared with 7 male normal controls of similar ages. We found reduced specific binding in the caudate nucleus and putamen in the alcoholic group, and the difference reached significance in the putamen. Specific binding was not decreased in the thalamus, which was examined as a reference structure. We also detected deficits in blood-to-brain transfer rate, K1, in the same regions of the alcoholic group, with a significant difference in the putamen. K1 was unchanged in the thalamus. The finding of reduced striatal VMAT2 in severe chronic alcoholic patients suggests that nigrostriatal monoaminergic terminals are reduced, with or without loss of neurons from the substantia nigra. The findings suggest that the damaging effects of severe chronic alcoholism on the central nervous system are more extensive than previously considered. PMID- 9749599 TI - Measurement of regional N-acetylaspartate after transient global ischemia in gerbils with and without ischemic tolerance: an index of neuronal survival. AB - We investigated the correlation between N-acetylaspartate (NAA) level and neuronal density in the hippocampal CA1 region of the brain after occlusion of both common carotid arteries for 5 minutes and reperfusion for 3 hours to 4 weeks in gerbils with and without ischemic preconditioning (tolerance). Animals were divided into four groups--the sham operated group, the nonpreconditioning (non-p) group, the single-preconditioning (single-p) group with 2-minute ischemia once 2 days before 5-minute ischemia, and the double-preconditioning (double-p) group with 2-minute ischemia twice 2 days before 5-minute ischemia (n = 6 for each group). The CA1 region was dissected out from freeze-dried sections for high performance liquid chromatographic assay of NAA, and adjacent sections were stained with cresyl violet for measurement of the neuronal density. Both NAA (pmol/microg dry weight) and the neuronal density (cells/mm) decreased in the non p group after 3 days (NAA = 24.0 +/- 3.0; neuronal density = 65 +/- 38 cells/mm) and 7 days (NAA = 17.9 +/- 2.5; neuronal density = 20 +/- 15 cells/mm) and in the single-p group after 7 days (26.4 +/- 3.0, 106 +/- 30) compared with the control group (NAA = 32.9 +/- 3.0; neuronal density = 203 +/- 9 cells/mm). There was no decrease in the double-p group. The NAA level and the neuronal density showed a good linear correlation. The regional NAA level may be used as an index of neuronal viability. PMID- 9749600 TI - Predictive factors for prolonged survival in acquired immunodeficiency syndrome associated progressive multifocal leukoencephalopathy. AB - Progressive multifocal leukoencephalopathy (PML) complicating the acquired immunodeficiency syndrome (AIDS) is typically inexorably progressive with death usually occurring within 6 months of symptom onset. Occasional patients have been observed to survive longer than 1 year, often with remission of clinical features. In this study, we identify predictive factors for prolonged survival in patients with biopsy proven, AIDS-associated PML, by comparing 7 patients with survival exceeding 12 months from symptom onset with 45 patients with shorter survivals. PML was the presenting manifestation of AIDS in 5 (71.4%) of 7 long term survivors compared with 8 (17.8%) of 45 short-term survivors. CD4 T lymphocyte counts were substantially higher in the long-term survivors, with 3 (42.9%) of 7 having counts exceeding 300 cells/mm3 in comparison with only 1 (4.3%) of 23 short-term survivors. Contrast enhancement on radiographic imaging was observed in 3 (50%) of 6 long-term survivors in comparison with 4 (8.9%) of 45 short-term survivors. Neurological recovery and radiographic improvement were not observed in any short-term survivors but were seen in 5 (71.4%) long-term survivors. There was no association between treatment modalities and survival. Predictors of long-term survival in AIDS patients with PML include PML as the heralding manifestation of AIDS, high CD4 T-lymphocyte count at disease onset, lesion enhancement on computed tomographic scan or magnetic resonance imaging, and evidence of recovery of neurological function. PMID- 9749601 TI - Acute inflammatory demyelinating polyradiculopathy in children: clinical and electrodiagnostic studies. AB - Clinical and electrophysiological features in 43 children with acute inflammatory demyelinating polyradiculoneuropathy (AIDP) were retrospectively studied. More than one-third of these children were less than 3 years old. Some distinctive clinical features specific to adults or to children were identified. Initial symptoms such as ataxia and severe limb or back pain were more frequent in children. By using the criteria suggested here, according to our neurophysiological findings, the diagnosis of AIDP could be proposed as early of the first days of illness in 90% of the children and is confirmed during the second week. The neurophysiological evolution was very similar in children and adults except that recovery occurred sooner in children. Prognosis was better in childhood (complete recovery in all but 2 patients with minor disabilities). PMID- 9749602 TI - Copper/zinc superoxide dismutase transgenic brain accumulates hydrogen peroxide after perinatal hypoxia ischemia. AB - Unlike the mature animal, immature mice transgenic for copper/zinc superoxide dismutase (SOD1) have greater brain injury after hypoxia-ischemia than their wild type nontransgenic littermates. To assess the role of oxidative stress in the pathogenesis of this injury, we measured histopathological damage, lipid peroxidation products, enzymatic activities of catalase and glutathione peroxidase, and hydrogen peroxide (H2O2) concentration in these animals before and after hypoxic-ischemic injury. Lipid peroxidation products were significantly increased 2 hours after the insult in both transgenic and nontransgenic brains in hippocampus, the most damaged brain region. Catalase activity did not increase in response to SOD1 overexpression or injury in either group. However, glutathione peroxidase activity, unchanged in response to overexpression, decreased significantly 24 hours after injury in both groups. At 24 hours after injury, greater H2O2 accumulation was observed in transgenic brains. Because SOD1 dismutates superoxide to H2O2, overexpression of SOD1 in the presence of developmentally low activities of the catalytic enzymes glutathione peroxidase and catalase leads to an increased production of H2O2, and may explain the increased brain injury observed after hypoxia-ischemia in neonatal SOD1 mice. PMID- 9749603 TI - Dominant negative effect of GTP cyclohydrolase I mutations in dopa-responsive hereditary progressive dystonia. AB - Hereditary progressive dystonia (HPD) is caused by the mutant gene encoding GTP cyclohydrolase I (GCH). The clinical presentation of this disease varies considerably, and many cases appear to be sporadic. We have previously proposed that this clinical variation may be due to differential expression of the mutant and normal GCH mRNA, presumably at the protein level. To provide support for this proposal, we studied a new Japanese family with HPD, in which 2 members were heterozygous for an exon-skipping mutation. This mutation produced truncated GCH, which shared 180-amino acid residues at the amino terminus of the normal enzyme (GCH180). An affected heterozygote had a higher mutant/normal mRNA ratio than an unaffected heterozygote, consistent with our previous finding in the HPD family with GCH114. A further study, using coexpression of the mutant with wild-type GCH in COS-7 cells, showed that three mutant GCHs inactivated the normal enzyme. GCH114 was most effective in enzyme inactivation, which was followed by GCH180 and a normally occurring mutant GCH209. These results suggested that the dominant negative effect of a mutant GCH on the normal enzyme might be one of the molecular mechanisms determining the heterogeneity of clinical phenotypes of HPD. PMID- 9749604 TI - Silent intracerebral microhemorrhages in patients with ischemic stroke. Amsterdam Vascular Medicine Group. AB - We compared the frequencies of signs of old intracerebral hemorrhages on brain magnetic resonance imaging scans in 66 patients with ischemic stroke, 69 with myocardial infarction, and 86 with peripheral arterial disease (a total of 221 patients). Magnetic resonance imaging scans were independently assessed by two investigators without knowledge of clinical or laboratory data. In 31 patients (14%) we found local cerebral hemosiderin deposits. In 24 patients they were clinically silent. Hemosiderin deposits were significantly more frequent in patients with ischemic stroke (26%) than in patients with myocardial infarction (4%) or peripheral arterial disease (13%). Hemosiderin deposits were associated with cerebral white matter lesions (odds ratio, 5.3; 95% confidence interval, 2.5 12.4). The odds ratios were higher in patients with severe cerebral white matter lesions. Our findings support the hypothesis that cerebral vessels of patients with ischemic stroke are more prone to rupture than those of patients with other manifestations of atherosclerotic disease, which may explain the higher incidence of intracerebral hemorrhages when these patients are treated with oral anticoagulants. The microhemorrhages were associated with cerebral white matter lesions, which suggests that they are another manifestation of cerebral small vessel disease. PMID- 9749605 TI - Acquired idiopathic generalized anhidrosis with isolated sudomotor neuropathy. AB - The pathogenesis and underlying lesion of acquired idiopathic generalized anhidrosis (AIGA) are apparently heterogeneous. We report a patient with AIGA in whom the eccrine glands were histologically normal. However, electron microscopic examination showed markedly low numbers of nerve terminals and unmyelinated axons associated with the eccrine glands. Our laboratory investigations suggest that degeneration of postganglionic sympathetic cholinergic nerve may be the underlying pathogenetic mechanism of anhidrosis in this patient. PMID- 9749606 TI - Overexpression of copper/zinc superoxide dismutase: a novel cause of murine muscular dystrophy. AB - Oxidative injury underlies the cellular injury and cell death in a variety of disease states. In muscular dystrophies, evidence from in vivo and in vitro studies suggests that muscle degeneration may be secondary to an increased susceptibility to oxidative stress. To address the role of free radical metabolism in the pathogenetic process of muscular dystrophies, we examined the muscle of transgenic mice that overexpress copper/zinc (Cu/Zn) superoxide dismutase. Overexpression of this enzyme can sensitize cells to oxidative injury, and Cu/Zn superoxide dismutase activity was elevated approximately fourfold above control levels in skeletal muscle of the transgenic strain. Examination of serum creatine phosphokinase levels in these mice revealed significant elevations after 2 months of age, indicative of active muscle breakdown. By 8 months of age, there was gross atrophy of the quadriceps muscle, and other hindlimb muscles were variably affected. Histologically, there was evidence of widespread muscle necrosis and regeneration, fiber splitting, and replacement of muscle with adipose and fibrous connective tissue, typical of a muscular dystrophy. Associated with the development of this degeneration was an increase in the levels of lipid peroxidation in the muscle of Cu/Zn superoxide dismutase transgenic mice, highlighting the central role of oxidative injury in this pathogenetic process. These results demonstrate that oxidative damage can be the primary pathogenetic process underlying a muscular dystrophy. PMID- 9749607 TI - Genetic association of two chromosome 14 genes (presenilin 1 and alpha 1 antichymotrypsin) with Alzheimer's disease. AB - We have investigated the association of two candidate genes on chromosome 14, presenilin 1 (PS1) and alpha1-antichymotrypsin (ACT), with the risk of sporadic Alzheimer's disease (AD) by using 427 AD cases and 250 controls. The frequency of the ACT*A allele was significantly higher in cases than controls (0.550 vs 0.466). The stratification of the ACT data by PS1 genotypes showed that the risk associated with the ACT*A allele was confined to PS1*1 carriers only. The two site haplotype data for PS1 and ACT showed that the A1 haplotype, carrying the ACT*A and PS1*1 alleles, was more frequent in cases than controls (0.310 vs 0.251), whereas the frequency of the T2 haplotype, carrying the ACT*T and PS1*2 alleles, was lower in cases than controls (0.177 vs 0.237). These data indicate a possible synergistic effect of these two loci on the risk of AD. PMID- 9749608 TI - Infrequent detection of human herpesvirus 6 DNA in peripheral blood mononuclear cells from multiple sclerosis patients. AB - Several studies have suggested an association between human herpesvirus 6 (HHV-6) infection and multiple sclerosis. As HHV-6 is predominantly a T-cell tropic virus, we examined the frequency of detection of HHV-6 genome in peripheral blood mononuclear cells from relapsing-remitting (n = 32) and chronic progressive (n = 14) patients and from healthy (n = 17) and neurological (n = 7) controls. Two sensitive polymerase chain reaction assays were used to target different regions within the HHV-6 genome. Depending on the polymerase chain reaction assay used, the detection of HHV-6 genome ranged from 11.7 to 23.5% (controls), 3.1 to 23.0% (relapsing-remitting), and 14.2 to 28.5% (chronic progressive). Although these observations do not exclude a pathogenic role for HHV-6 in multiple sclerosis, they indicate a lack of correlation between HHV-6 infection of peripheral blood mononuclear cells and the development of multiple sclerosis. PMID- 9749609 TI - Clinical and genetic evaluation of a family with a mixed dystonia phenotype from South Tyrol. AB - The gene causing early-onset torsion dystonia (DYT1) has recently been identified, and two new dystonia genes, one for adult-onset focal dystonia (DYT7) and one for a mixed dystonia phenotype (DYT6), have been mapped. We evaluated clinically a family from South Tyrol (Northern Italy) with 6 definitely affected individuals who display an unusually large phenotypic range of dystonic symptoms. We excluded the GAG deletion in the DYT1 gene and linkage to any of the above mentioned dystonia loci, thus suggesting an as yet undefined dystonia gene in our family. PMID- 9749610 TI - Somatotopical organization of striatal activation during finger and toe movement: a 3-T functional magnetic resonance imaging study. AB - The present study aimed at determining the distribution and somatotopical organization of striatal activation during performance of simple motor tasks. Ten right-handed healthy volunteers were studied by using a 3-T whole-body magnetic resonance unit and echo planar imaging. The tasks consisted of self-paced flexion/extension of the right fingers or toes. Motor activation was found mainly in the putamen posterior to the anterior commissure (10 of 10 subjects) and the globus pallidus (6 subjects), whereas the caudate nucleus was activated in only 3 subjects, and in a smaller area. Thus, performance of a simple motor task activated the sensorimotor territory of the basal ganglia. Within the putamen, there was a somatotopical organization of the foot and hand areas similar to that observed in nonhuman primates. These data suggest that functional magnetic resonance imaging can be used to study normal function of the basal ganglia and should therefore also allow investigation of patients with movement disorders. PMID- 9749611 TI - Evoked potential abnormality scores are a useful measure of disease burden in relapsing-remitting multiple sclerosis. AB - Fifty patients with relapsing-remitting multiple sclerosis were examined and studied with serial evoked potential and magnetic resonance imaging (MRI) measurements as part of a clinical trial. An evoked potential abnormality score (EPAS) for each testing session was calculated consisting of the total number of abnormal tests. The EPAS correlated well with Expanded Disability Status Scale (EDSS) at years 0, 1, and 2, with Spearman correlation coefficient scores of 0.68, 0.66, and 0.72, respectively. MRI lesion volume correlations ranged from 0.27 to 0.34 for the EDSS. EPAS are a potentially useful surrogate measure of clinical disability in relapsing-remitting multiple sclerosis. PMID- 9749612 TI - Correlation between cytomegalovirus infection and IgM anti-MAG/SGPG antibody associated neuropathy. AB - IgM anti-myelin-associated glycoprotein (anti-MAG)/sulfated glucuronyl paragloboside (SGPG) antibody is found in some patients with chronic polyneuropathy (CP). An antigen-driven process is considered to induce this autoantibody, but the agent has yet to be identified. It has been reported that sera from cytomegalovirus (CMV)-infected patients contained anti-SGPG antibody. To clarify the mechanism of the production of the anti-MAG/SGPG antibody, we investigated CMV DNA in sera from 26 patients with IgM anti-MAG/SGPG antibody positive CP. Twenty-three (88%) had CMV DNA. The positive frequency was significantly higher than the frequencies in sera from patients with IgM anti MAG/SGPG-negative CP, the other disease controls, and the normal control subjects. There were no statistical differences in the frequencies of Epstein Barr virus DNA between anti-MAG/SGPG-positive and anti-MAG/SGPG-negative CP and between anti-MAG/SGPG-positive CP and each disease control. Moreover, no herpes simplex virus 1 DNA was detected in the sera from patients with anti-MAG/SGPG positive CP. The strong correlation of anti-MAG/SGPG-positive CP with the presence of serum CMV DNA suggests that CMV infection induces the IgM anti MAG/SGPG antibody. PMID- 9749613 TI - Cerebrospinal fluid F2-isoprostane levels are increased in Alzheimer's disease. AB - Postmortem studies have associated Alzheimer's disease (AD) with regionally increased oxidative damage to brain. Lacking, however, is a specific marker of oxidative damage to brain that may be measured during life. We tested the hypothesis that cerebrospinal fluid (CSF) concentrations of F2-isoprostanes (F2 IsoPs), stable products of arachidonate peroxidation, are increased in CSF of AD patients. CSF from lateral ventricles (VF) was analyzed from 11 AD patients and 11 control subjects who participated in a rapid autopsy program. VF F2-IsoP concentrations were significantly elevated in AD patients compared with control subjects (72 +/- 7 vs 46 +/- 4 pg/ml) and were significantly linearly correlated with brain weight (-0.3 pg/ml/g, r2 = 0.32). These results suggest that quantification of CSF F2-IsoP concentrations may provide a useful biomarker of central nervous system oxidative damage in AD. PMID- 9749614 TI - Search for varicella zoster virus in giant cell arteritis. AB - Polymerase chain reaction and immunohistochemical analyses of formalin-fixed temporal arteries from 10 pathologically verified cases of giant cell arteritis did not reveal varicella zoster virus antigen or DNA. PMID- 9749616 TI - Presence of the anti-Jo-1 autoantibody excludes inclusion body myositis. PMID- 9749615 TI - Glial cytoplasmic inclusions in white matter oligodendrocytes of multiple system atrophy brains contain insoluble alpha-synuclein. AB - Recently, alpha-synuclein was shown to be a structural component of the filaments in Lewy bodies (LBs) of Parkinson's disease (PD), dementia with LBs (DLB) as well as the LB variant of Alzheimer's disease, and this suggests that alpha-synuclein could play a mechanistic role in the pathogenesis of these disorders. To determine whether alpha-synuclein is a building block of inclusions in other neurodegenerative movement disorders, we examined brains from patients with multiple system atrophy (MSA) and detected alpha-synuclein, but not beta- or gamma-synuclein, in glial cytoplasmic inclusions (GCIs) throughout the MSA brain. In MSA white matter, alpha-synuclein-positive GCIs were restricted to oligodendrocytes, and alpha-synuclein was localized to the filaments in GCIs by immunoelectron microscopy. Finally, we demonstrated that insoluble alpha synuclein accumulated selectively in MSA white matter with alpha-synuclein positive GCIs. Taken together with evidence that LBs contain insoluble alpha synuclein, our data suggest that a reduction in the solubility of alpha-synuclein may induce this protein to form filaments that aggregate into cytoplasmic inclusions, which contribute to the dysfunction or death of glial cells as well as neurons in neurodegenerative disorders with different phenotypes. PMID- 9749617 TI - Increased delivery of nerve growth factor to neuronal cell body reduces up regulation of genes. PMID- 9749618 TI - Imaging and multiple sclerosis. PMID- 9749619 TI - CD95 (APO-1/Fas) and Parkinson's disease. PMID- 9749620 TI - Factor V Leiden as a cause of hemiplegic cerebral palsy, neonatal stroke, and placental thrombosis? PMID- 9749621 TI - Pocket Monsters, a popular television cartoon, attacks Japanese children. PMID- 9749622 TI - Pocket Monsters attacks Japanese children via media. PMID- 9749623 TI - Development of monoclonal antibodies specifically recognizing the cyst stage of Entamoeba histolytica. AB - Protozoan cysts were isolated according to a two-step sucrose gradient procedure. Pure cysts of Entamoeba histolytica, in fixed and native states, were injected into BALB/c mice intraperitoneally for immunization. The spleens of these animals were used for fusion with AG8 mouse myeloma cells. Hybridomas were obtained and tested for the recognition of E. histolytica, E. dispar, E. coli, E. hartmanni, Endolimax nana, Jodamoeba biitschlii, and Giardia lamblia. Three monoclonal antibodies were identified that reacted only with cysts and trophozoites of E. histolytica. These can be used for differentiation and identification of E. histolytica in feces. PMID- 9749624 TI - Immunoglobulin M-capture biotin-streptavidin enzyme-linked immunosorbent assay for detection of antibodies to dengue viruses. AB - A biotin-streptavidin system was adapted to an IgM-capture ELISA for detection of dengue antibodies in human sera. To develop this assay, high titers of antibodies to flavivirus were purified by ion-exchange chromatography (DEAE-cellulose) and labeled with biotin. Heavy chain-specific goat anti-human IgM was first bound to the wells of a polystyrene microtiter plate, followed by binding of IgM in test specimens, and the use of tetravalent dengue antigens (dengue 1-4), biotin labeled anti-flavivirus IgG, and streptavidin-peroxidase conjugate. The sensitivity and specificity of the IgM-capture biotin-streptavidin ELISA (IgM-BS ELISA) in acute sera were 83.3% of patients with dengue infection and 95.3% of nondengue-infected cases, respectively. The positive predictive value was 92.4% and the negative predictive value was 89.2%. The efficiency of test was 90.4%. In convalescent sera, the sensitivity and specificity of IgM-BS-ELISA were 100% and 92.6%, respectively. The predictive values of positive and negative results were 90.3% and 100%, respectively. The efficiency of test was 95.6%. The agreement rate of IgM-BS-ELISA and standard hemagglutination inhibition test was good: kappa (kappa) values were 0.79 for acute sera and 0.91 for convalescent sera. The correlation between two methods was quite good, with correlation coefficients (r) of 0.76 for acute sera and 0.85 for convalescent sera (P < 0.001). The results indicate that the IgM-BS-ELISA is highly sensitive, specific, simple to perform, and rapid. PMID- 9749625 TI - Identification of Brazilian flaviviruses by a simplified reverse transcription polymerase chain reaction method using Flavivirus universal primers. AB - We report a simplified reverse transcription-polymerase chain reaction (RT-PCR) method for identification of Brazilian flaviviruses based on the patterns of electrophoretic separation of the amplicons. The RT-PCR was done on the culture fluids of Aedes albopictus C6/36 cells infected with Brazilian flaviviruses, without previous extraction of viral RNA, using Flavivirus universal primers that anneal to highly conserved sequences within the nonstructural protein 5 and 3'- non translated region of the virus genome. Genomes of 13 Brazilian Flavivirus isolates were amplified. It was not possible to amplify the genome of Bussuquara virus. Analysis of the RT-PCR products gave reproducible results and three distinct amplicon patterns were observed. Cacipacore (800-850 basepairs [bp]) and yellow fever viruses (600 bp) yielded a single amplicon; dengue virus types 1 and 2 (650 and 550 bp), dengue virus type 4 (550 and 450 bp), Iguape (650-600 bp and 750-700 bp), St. Louis encephalitis (700 and 650-600 bp), and Rocio viruses (600 and 500-550 bp) yielded two amplicons; and Ilheus virus yielded five amplicons, two larger than 1,000 bp, one 650-700 bp, one 550-600 bp, and one 450-500 bp. The analysis of amplicon DNA sequences of six viruses showed homology with the 3'- nontranslated region of Flavivirus genome. The use of the Flavivirus universal primers in this simple RT-PCR technique is suitable as a screening test for the genus Flavivirus, with the exception of Bussuquara virus, in Brazilian isolates in tissue culture fluid. PMID- 9749626 TI - Serodiagnosis and epidemiology of visceral leishmaniasis in Turkey. AB - Infantile Mediterranean visceral leishmaniasis (IVL) and anthroponotic cutaneous leishmaniasis (ACL) have long been known to exist in the western and southeastern Turkey, respectively. To further study these and other related diseases, a recombinant antigen (rK39) specific to VL was used in an ELISA for serodiagnosis of selected patients and for screening dog reservoir populations in several endemic sites. Among 24 confirmed VL cases from western Turkey, the rK39 ELISA proved to be more sensitive than a combination of cultivation and microscopy of bone marrow aspirates. The specificity of rK39 for leishmaniasis was demonstrated by its lack of cross-reactivity with sera from other human diseases in the same sites. Interestingly, six of the 83 parasitologically proven ACL cases from southeast Turkey were also rK39 positive. The end point titers of the positive VL and CL cases vary from 10(-2) to 10(-5) and from 10(-2) to 10(-3), respectively. The rK39 ELISA was also used to screen 494 apparently healthy dogs from Urfa in southeast Turkey, Manisa/Alasehir near the Aegean Sea, and Karabuk near the Black Sea. Eighteen rK39-positive cases (3.6%), all from the latter two areas, were found to have varying endpoint titers (10(-2)-10(-4)). The high titers predicted increased severity and frequency of the clinical symptoms (i.e., lymphadenopathy, depilation, skin lesion, weight loss and/or death), which were manifested subsequently in 16 of these 18 cases. In addition, more positive canine cases were diagnosed by the rK39 ELISA preclinically than the procedures to detect parasites postsymptomatically in the lymph node aspirates. The use of the rK39 ELISA as a sensitive tool makes it possible to demonstrate coendemicity of canine and human VL, as expected in the case of IVL. The results also point to the possible presence of additional VL types in western Turkey and cutanovisceral type in the southeast part of this country. PMID- 9749627 TI - Variations in fecal Schistosoma japonicum egg counts. AB - Variations in fecal Schistosoma japonicum egg counts were studied in ZhuXi administrative village, JiangXi Province, China. Population stool examinations were collected with duplicate, standard, 41.5-mg Kato-Katz thick smears on seven consecutive days for 570 individuals from two natural (individual) villages: village I with high endemicity and village II with low endemicity. The proportion of individuals with at least one positive count increased from 42.4% after a single measurement to 68.3% after seven measurements in village I (n = 356), and from 17.0% to 36.0% in village II (n = 214), respectively. This demonstrates a very high variation in repeated S. japonicum egg counts and a considerable lack of sensitivity of the Kato-Katz technique; light and moderate infections are especially missed with a single or a few measurements. The observed day-to-day variation in individual egg counts is highly aggregated (variance higher than the mean) and suggestive of a negative binomial distribution. For five individuals on three days, repeated sampling from different locations of a stool specimen shows a clear trend with egg counts decreasing from the beginning of the stool to the end and from the outside layer to the center. Ten multiple samples from a particular subsection (10-30 g) of a stool specimen for 44 positive individuals still showed aggregation in egg counts, particularly for high intensities of infection. This means that the aggregation in repeated daily S. japonicum egg counts cannot be explained alone by a specific day-to-day component and variation in the concentration of eggs at different locations in the stool. There also exists clustering of eggs within parts of the stool. PMID- 9749628 TI - Use of antimalarial drugs in Mali: policy versus reality. AB - Inappropriate use of antimalarial drugs undermines therapeutic efficacy and promotes the emergence and spread of drug-resistant malaria. Strategies for improving compliance require accurate information about current practices. Here we describe Knowledge-Attitude-Practice surveys conducted among health providers and consumers in two Malian villages, one rural and one periurban. All sanctioned providers limited their first choices of antimalarial drug to those recommended by the national malaria control program and reported using correct dosing regimens. However, the majority of consumers in the two villages chose non recommended treatments for malaria and reported suboptimal treatment regimens when they did use recommended drugs. Antimalarial drugs were also widely available from unsanctioned sources, often accompanied by erroneous advice on dosing regimens. This study demonstrates that even when the most peripheral health providers are well-trained in correct use of antimalarial drugs, additional measures directly targeting consumers will be required to improve drug use practices. PMID- 9749630 TI - Failure of penicillin treatment of yaws on Karkar Island, Papua New Guinea. AB - The endemic treponematosis yaws remains a significant cause of morbidity in many tropical countries, despite mass treatment campaigns to eradicate it. An outbreak of yaws in Marup village on Karkar Island, Papua New Guinea in 1988 provided an opportunity to monitor the outcome of treatment with penicillin over an extended period. Thirty-nine children with clinical yaws (6% of 632 examined) were monitored clinically and serologically, for nearly two years after mass treatment of all villagers with the World Health Organization recommended dosages of benzathine penicillin. Lesions resolved within one month of treatment in all but four (10%) children, three of whom were initially successfully retreated. Before treatment, the Venereal Disease Research Laboratory (VDRL) test result was reactive in 67% of the children and treponema-specific IgM antibody test results were reactive in 41%. Within six months of treatment, of those reactive, the VDRL titer decreased significantly in 25 (96%) of 26 and IgM antibody test results became negative in 13 (81%) of 16 children. However, by the end of follow-up, 11 (28%) of the 39 children had developed clinical and/or serologic evidence of relapse. In these children, response to further treatment was slow and, in three, evidence of active infection persisted or recurred, despite repeated courses. Exogenous reinfection was unlikely in this isolated community, in which the occurrence of yaws was closely monitored after universal treatment. Treatment failure was most likely to have been due to reduced susceptibility to penicillin of Treponema pallidum subsp. pertenue. PMID- 9749631 TI - Somatosensory discrimination deficits following pediatric cerebral malaria. AB - Pathologic studies of central nervous system damage in human falciparum malaria indicate primary localization in the cerebral white matter. We report a sensory perceptual investigation of 20 Ghanaian children with a recent history of cerebral malaria who were age-, gender-, and education-matched with 20 healthy control subjects. Somatosensory examinations failed to show any evidence of hemianesthesia, pseudohemianesthesia, or extinction to double simultaneous tactile stimulation. While unilateral upper limb testing revealed intact unimanual tactile roughness discrimination, bimanual tactile discrimination, however, was significantly impaired in the cerebral malaria group. A strong negative correlation (r = -0.72) between coma duration and the bimanual tactile roughness discrimination test was also found. An inefficiency in the integrity of callosal fibers appear to account for our findings, although alternative subcortical mechanisms known to be involved in information transfer across the cerebral hemispheres may be compromised as well. PMID- 9749629 TI - Reduction of fecal contamination of street-vended beverages in Guatemala by a simple system for water purification and storage, handwashing, and beverage storage. AB - Street-vended foods and beverages, an integral part of urban economies in the developing world, have been implicated in cholera transmission in Latin America. To improve the microbiologic quality of market-vended beverages in Guatemala, we tested a simple system consisting of dilute bleach (4.95% free available chlorine) for water purification, narrow-mouth plastic vessels with spigots for disinfecting and storing water and for preparing and storing beverages, handwashing soap, and education in using the system. We conducted a randomized controlled intervention trial among 41 vendors who received the intervention and 42 control vendors, comparing total and fecal coliform bacteria and Escherichia coli contamination of market-vended beverages, stored water, and vendors' hands. Samples were obtained at baseline and at each of six weekly follow-up visits. At baseline, fecal coliform bacteria were found in 40 (48%) market-vended beverages and E. coli in 14 (17%). When compared with samples from control vendors, a significant decrease in total coliform (P < 0.001) and fecal coliform (P < 0.001) bacteria in samples of stored water and beverages sold by intervention vendors was observed over the course of the study. The vessel system was well accepted by vendors. This simple inexpensive system consisting of hypochlorite disinfectant, plastic vessels, soap, and education can significantly reduce fecal contamination of market-vended beverages. PMID- 9749632 TI - High serum calcium in human brucellosis: a case-control study. AB - In a retrospective case-control study of 58 cases of human brucellosis, adjusted mean serum calcium levels were found to be significantly higher in patients with brucellosis compared with controls: mean (95% confidence interval) = 2.39 (2.35 2.42) mmol/L versus 2.30 (2.26-2.34) mmol/L (P = 0.0012). The possible mechanisms underlying the cause of hypercalcemia in human brucellosis are discussed. PMID- 9749633 TI - Ultrasonographic detection of living adult Wuchereria bancrofti using a 3.5-MHz transducer. AB - Adult Wuchereria bancrofti can be readily detected by ultrasound in the lymphatic vessels of the spermatic cord with a 7.5-MHz transducer, but most ultrasound machines in developing countries are equipped with 3.5-MHz transducers. To assess the potential for ultrasound as a tool for diagnosis and epidemiologic assessment in lymphatic filariasis, we compared the performance of 3.5-MHz and 7.5-MHz transducers in 61 men in Recife, Brazil. All men had three ultrasound examinations using a 3.5 MHz transducer and an examination with a 7.5-MHz probe. Using the 7.5-MHz transducer, adult W. bancrofti were detected in 41 men; 81 adult worm nests were detected. Sixty-four (79%) nests were detected with the 3.5 MHz probe, each on all three examinations. The 3.5-MHz probe correctly identified 35 (85.4%) of 41 men as infected; sensitivity increased with lymphatic vessel diameter. Ultrasonographic examination with a 3.5-MHz transducer is a sensitive method for detection of adult W. bancrofti in men and merits consideration as a tool for rapid epidemiologic assessment. PMID- 9749634 TI - Adult Toxocara cati infections in U.S. children: report of four cases. AB - We report four cases of passage of subadult or adult Toxocara cati worms by young children ages 20 months to seven years. Worms were expelled rectally in two cases and in two cases they were vomited. A single worm was passed in two cases, three worms in one case, and 15 worms in the fourth case. All worms that were available for study were identified as T. cati by morphologic criteria, including the arrow shaped cervical alae and the digitiform shape of the male tail. None of the four children exhibited clinical signs of ocular or visceral larva migrans, and in two cases where serum samples were available, neither child had a titer to Toxocara. These results further the argument that these children acquired the worms through the ingestion of immature worms passed by infected cats, not through the ingestion of infective eggs. Although the children were generally not ill as a result of these unusual infections, it does serve to reinforce the public health issue that potential serious consequences can occur where children have exposure to an environment that has been contaminated with cat feces, or, more specifically, infective eggs, and could become infected with larval forms of Toxocara. PMID- 9749636 TI - Transfection and heat-inducible expression of molluscan promoter-luciferase reporter gene constructs in the Biomphalaria glabrata embryonic snail cell line. AB - Studies were initiated to begin developing a genetic transformation system for cells derived from the freshwater gastropod, Biomphalaria glabrata, an intermediate host of the human blood fluke Schistosoma mansoni. Using a 70-kD heat-shock protein (HSP70) cDNA probe obtained from the B. glabrata embryonic (Bge) cell line, we cloned from Bge cells a complete HSP70 gene including a 1-kb genomic DNA fragment in its 5'-flanking region containing sequences indicative of a HSP promoter. Identified in the 5'-half (416 nucleotides) of this genomic fragment were TATA and CAAT boxes, two putative transcription initiation sites, and a series of palindromic DNA repeats with shared homology to the heat-shock element consensus sequence (Bge HSP70(0.5k) promoter). The 3'-half of this upstream flanking region was comprised of a 508-base intron located immediately 5' of the ATG start codon. To determine the functionality of the putative snail promoter sequence, Bge HSP promoter/luciferase (Luc) reporter gene constructs were introduced into Bge cells by N-(1-(2,3-dioleoyloxy) propyl)-N,N,N trimethylammonium methylsulfate (DOTAP)-mediated transfection methods, and assayed for Luc activity 48 hr following a 1.5-hr heat-shock treatment (40 degrees C). Compared with control vectors or the Bge HSP70(0.5k/1.0k) promoter constructs at 26 degrees C, a 10- to 300-fold increase in Luc expression was obtained only in the Bge HSP70 promoter/Luc-transfected cells following heat shock. Results of transfection experiments demonstrate that the Bge HSP70(0.5k) DNA segment contains appropriate promoter sequences for driving temperature inducible gene expression in the Bge snail cell line. This report represents the first isolation and functional characterization of an inducible promoter from a freshwater gastropod mollusc. Successful transient expression of a foreign reporter gene in Bge cells using a homologous, inducible promoter sequence now paves the way for development of methods for stable integration and expression of snail genes of interest into the Bge cell line. PMID- 9749635 TI - The public health significance of urinary schistosomiasis as a cause of morbidity in two districts in Mali. AB - Schistosoma haematobium-related morbidity was studied in the perennial irrigation area of Office du Niger and the small reservoirs area of Plateau Dogon in Mali. Questionnaire, clinical, parasitologic, and ultrasound examination data were collected from 1,041 individuals at the baseline survey in 1991; 705 were re examined one year after treatment. At baseline, the overall prevalence of S. haematobium infection was 55.2%; half of those infected had no clinical symptoms and 30% had pathologic lesions. Both infection and morbidity were more frequent in children than in adults, with a peak prevalence at 7-14 years of age. The rates of lesions were more than twice as high in those heavily infected as in lightly infected individuals. Reagent strip testing for microhematuria was more sensitive in detecting individuals with pathologic lesions than in detecting individuals with infection. One year after treatment with praziquantel, more than 80% of the urinary tract lesions had cleared. It is concluded that S. haematobium related morbidity is frequent in Mali, but passive case detection for treatment would not cover a great deal of early stages of the disease; active intervention using reagent strip testing for microhematuria at the most peripheral levels would be an efficient system for morbidity control and monitoring of control operations. PMID- 9749637 TI - Transformation, development, and transmission of axenically cultured amastigotes of Leishmania mexicana in vitro and in Lutzomyia longipalpis. AB - Axenic cultures of Leishmania mexicana amastigotes were transformed to promastigotes in vitro and in vivo in Lutzomyia longipalpis. In vitro, both exponential phase and stationary phase amastigotes were capable of transforming and growing as promastigotes, but exponential phase amastigotes completed this transition more quickly. In vivo, both populations were capable of establishing infections in sand flies by membrane feeding and could be transmitted to BALB/c mice via bite. A variety of morphologic forms could be observed in vivo, including putative metacyclic promastigotes. Infection rates in sandflies with axenic amastigotes were comparable with those achieved with lesion-derived amastigotes, supporting the use of these cultured forms in studies of parasite biology. PMID- 9749638 TI - Anti-leishmanial IgE antibodies: a marker of active disease in visceral leishmaniasis. AB - Visceral leishmaniasis (VL) is characterized by a depression of the T helper cell type 1 immune response. Although mRNA expression for interleukin-4 (IL-4) is observed, evidence of the role of this cytokine in the pathogenesis of VL has been lacking. Since IL-4 is involved in IgE synthesis, we measured the total IgE and Leishmania antigen-specific IgE antibody levels in sera from patients with VL. Specific IgE antibodies detected by an ELISA technique after absorbing the sera with purified sheep IgG anti-human IgG were found in all 23 patients with VL and were not detected in subjects with subclinical Leishmania chagasi infection (n = 10), Chagas' disease (n = 10), atopic patients (n = 10), and healthy controls (n = 10). Levels of Leishmania-specific IgE (optical density values) before and after treatment were 0.100 +/- 0.03 (mean +/- SD) and 0.028 +/- 0.002, respectively (P < 0.05). These results indicate that a specific IgE response is useful in the diagnosis of active disease and to evaluate response to treatment. PMID- 9749639 TI - Serum IgM antibody response to the galactose-inhibitable adherence lectin of Entameoba histolytica. AB - An ELISA for detection of serum IgM antibodies to the galactose-inhibitable adherence lectin of Entamoeba histolytica revealed that 2.8% of uninfected controls, 0.0% of controls infected with other parasites, 13.4% of asymptomatic amebic infections, 55% of colitis patients, and 77% of amebic liver abscess patients from Cairo, Egypt and Durban, South Africa had serum anti-lectin IgM antibodies. Of acute amebic colitis patients with symptoms for less than one week, only 6% possessed serum IgG anti-lectin antibodies, yet 45% had serum IgM antibodies to the amebic lectin. This compares with 65% of sera in acute colitis patients positive for lectin antigen as determined by ELISA with anti-lectin monoclonal antibodies. In conclusion, an ELISA for serum anti-lectin IgM antibodies appears to have greater clinical utility in the setting of acute amebic colitis than an ELISA for anti-lectin IgG antibodies, but is no more sensitive than an ELISA for detection of lectin antigen in sera. PMID- 9749640 TI - Cryptosporidiosis in urban Zambian children: an analysis of risk factors. AB - In four crowded townships of Lusaka, Zambia, the prevalence of cryptosporidiosis in 222 children with diarrhea was 18%, with marked temporal and geographic variation over the course of one rainy season. Using data on the finding of oocysts of Cryptosporidium parvum in urban water supplies, the areas under study were categorized as high or low risk. Prevalence of cryptosporidiosis in children with diarrhea was higher in high risk areas after stratification by early/late stage of the rains (Mantel-Haenszel odds ratio [OR] = 2.9, 95% confidence interval [CI] = 1.3, 6.7; P = 0.008). Cryptosporidiosis was not associated with keeping animals, nutritional status, or parental education, but was apparently more common in breast fed children (OR = 2.7, 95% CI = 1.1, 6.9; P = 0.01), although the proportion of exclusively breast fed children was not measured. Since most of these infections were of short duration, we conclude that transmission of C. parvum can vary dynamically within one city and over short periods of time, and that water-borne contamination may be a substantial influence. PMID- 9749642 TI - Ecology and demographics of hantavirus infections in rodent populations in the Walker River Basin of Nevada and California. AB - To study the ecologic correlates of hantavirus in deer mice (Peromyscus maniculatus), we sampled 114 sites in the Walker River Basin of Nevada and California in 1995-1996. Blood samples were tested for antibody to hantavirus, and a subset of samples was also tested for virus RNA by reverse transcription polymerase chain reaction. Average prevalence of antibody-positive mice was 17%, with heavier males the most likely to be infected. Antibody prevalence varied within repeatedly sampled sites from 0% to 50% over the course of several months, suggesting possible infection cycles. Although there was no linear correlation between deer mouse density and antibody prevalence on sample sites, more complex relationships between density and prevalence appeared likely. Specifically, infections were less likely where rodent densities were lower than a critical threshold value. However, above this value, density had no effect on prevalence. PMID- 9749641 TI - High prevalence of hantavirus infection in Indian communities of the Paraguayan and Argentinean Gran Chaco. AB - Serologic evidence of past infection with a Sin Nombre-like hantavirus(es) was demonstrated in 78 (40.4%) of 193 Indians living in western Paraguay and in 38 (17.1%) of 222 Indians inhabiting the Salta province of northern Argentina. In both populations seroprevalence increased with age, with the most striking increase occurring at 18 years of age in the Paraguayan population and at 35 years of age in the Salta population. The peak prevalences in both populations (66.6% and 44.0%, respectively) were seen in Indians > 53 years old. Although no sex difference was observed in the Paraguayan Indians, in the Salta population seroprevalence was greater in males than in females. Familiar clustering of the infection was observed. The data indicate that the Indian populations of the Gran Chaco are frequently exposed to and survive infection with a Sin Nombre-like virus(es). Possible explanations of this novel epidemiology are discussed. PMID- 9749643 TI - Mayaro virus fever in French Guiana: isolation, identification, and seroprevalence. AB - This paper reports the first isolation of Mayaro (MAY) virus from a patient infected in French Guiana. The identification was initially performed using immunofluorescent antibody testing with specific mouse antibody, and confirmed by plaque-reduction neutralization testing and reverse transcription-polymerase chain reaction. To determine if MAY virus infection is widespread in French Guiana, a serosurvey was performed to determine the prevalence of antibody to this virus in various ethnic groups and areas of French Guiana. Human sera (n = 1,962) were screened using the hemagglutination inhibition (HI) test. To determine whether MAY virus circulates in the rain forest, a serosurvey in monkey populations was performed. Monkey sera (n = 150) were also screened for antibody to MAY virus using HI testing. Of the human sera tested, 6.3% were positive for anti-MAY virus antibodies. Significant differences in MAY virus seroprevalence between different age groups were observed. Seroprevalence rates increased with age, with a large increase in people 10-19 years of age in comparison with those less than 10 years of age. After adjustment for age, significant differences were also found between places of residence. The prevalence of anti-MAY virus antibody was higher in people living in contact with the forest, especially in the Haut Oyapock area (odds ratio [OR] = 97.7, 95% confidence interval [CI] = 48.2-197.9) and along the Maroni River (OR = 39.7, 95% CI = 20.6-76.6). The ethnic differences observed in this study were probably due to differences in residence. Among monkeys, higher seroprevalence rates were found in Alouatta seniculus (66.0%) than in Saguinus midas (18.2%). Among Alouatta, the seroprevalence increased significantly with weight (and therefore with age). This study indicates that MAY virus is present in French Guiana, and human infections occur in areas where people live near the tropical rain forest. PMID- 9749644 TI - The 1993 dengue 2 epidemic in North Queensland: a serosurvey and comparison of hemagglutination inhibition with an ELISA. AB - An epidemic of dengue type 2 infection occurred in North Queensland during 1992 and 1993. A random serosurvey of 1,000 residents of a population that experienced this epidemic only during 1993 was conducted to determine the proportion of the population at risk for secondary infection in the event of another epidemic with a different serotype. The ability of an ELISA to detect prior exposure to the dengue virus was compared with the hemagglutination inhibition assay. Dengue 2 virus plaque-reduction neutralization assays were performed to evaluate the specificity of the antibody response. Antibodies to dengue virus, or closely related flaviviruses, were detected in 61.9%. Seroprevalence increased with age and correlated well with known previous epidemics in the region. The sensitivity and specificity of the ELISA was 99.2% and 96.2%, respectively. An estimated 26% of the population was infected during the 1993 epidemic. PMID- 9749645 TI - GB virus C/hepatitis G virus infection among Colombian native Indians. AB - To elucidate the prevalence of GB virus C/hepatitis G virus (GBV-C/HGV) infection in Colombian native Indians, serum GBV-C/HGV RNA was assayed in 163 native Indians and 67 members of the general population in Colombia. The native Indians (males:females = 40:123) and the members of the general population (males:females = 20:47) were tested by reverse transcription-semi-nested polymerase chain reaction. Of the 163 native Indians, 10 (6.1%) were positive for GBV-C/HGV RNA, compared with one (1.5%) of 67 from the general population. All Indians were negative for hepatitis B surface antigen and antibody to hepatitis C virus. Of 10 Indians with GBV-C/HGV RNA, the genotype of nine subjects was the Asian type. These data indicated that 1) the prevalence of GBV-C/HGV RNA in Colombian native Indians is high, and 2) GBV-C/HGV was probably brought from Asia and inherited for generations in some native Indian groups. PMID- 9749646 TI - Development and laboratory evaluation of a polymerase chain reaction for monitoring Schistosoma mansoni infestation of water. AB - A sensitive and specific detection of cercariae of human schistosomes is required for better definition of risk of infection. In the present study, we have developed a polymerase chain reaction (PCR) assay for the detection of cercariae of Schistosoma mansoni in water. A simple DNA preparation was adapted for this purpose, and PCR primers were designed based on the 121-basepair highly repeated sequence we previously identified in the genome of S. mansoni. The PCR assay detected as little as 10(-6) ng of S. mansoni DNA, and the high sensitivity enabled the detection of a single cercaria. For trapping of cercariae we adapted a filtration apparatus previously used for separating schistosome eggs from turbid enzymatic digests of tissues. A single cercaria could be detected in repeated tests of water filtrates. Since the target DNA is tandemly arranged, a ladder pattern of the PCR products was demonstrated. A direct relationship was demonstrated between the number of ladder bands of the amplification products, and DNA concentration or number of cercariae. The feasibility of semiquantitation of schistosome larvae in natural water was thus suggested. The potential of the procedures described here for epidemiologic studies is discussed. PMID- 9749647 TI - Allelic diversity at the merozoite surface protein-1 locus of Plasmodium falciparum in clinical isolates from the southwestern Brazilian Amazon. AB - Nucleotide sequences of each variable block in the Plasmodium falciparum merozoite surface protein-1 gene (PfMSP-1) may be grouped into one of two or three possible allelic types, named after the reference isolates MAD20, K1, and RO33. Allelic diversity at this locus basically results from different combinations of allelic types in variable blocks. We used a polymerase chain reaction (PCR)-based strategy to type the variable blocks 2, 4a, 4b, and 10 of the PfMSP-1 gene of P. falciparum isolates from 54 symptomatic malaria patients living in Rondonia, a hypoendemic area in the southwestern Brazilian Amazon. Ten different PfMSP-1 gene types, defined as unique combinations of allelic types in variable blocks, were identified among the 54 isolates. Twenty-one isolates (39%) harbored more than one gene type and two had at least three genetically distinct clones. Hybrid sequences, with a MAD20-type sequence in the 5' segment (4a) and a K1-type sequence in the 3' segment (4b), were quite common in block 4. Direct sequencing of block 4 PCR products revealed a new putative recombination site in four isolates. In contrast with previous studies, the observed distribution of gene types does not deviate significantly from that expected under the null hypothesis of random association between allelic types detected in each variable block. These contradictory data are discussed with reference to the immunoepidemiologic features prevailing in distinct malaria-endemic areas. PMID- 9749648 TI - Gametocyte infectivity by direct mosquito feeds in an area of seasonal malaria transmission: implications for Bancoumana, Mali as a transmission-blocking vaccine site. AB - Infectivity of gametocytemic volunteers living in Bancoumana, a village 60 km from Bamako, Mali, was determined by direct feeds of laboratory-reared Anopheles gambiae s. l. Gametocytemic adolescents (10-18 years old) were as infectious to mosquitoes as younger volunteers and appear to be a more suitable population for testing transmission-blocking efficacy as compared with adults (> 18 years old). To begin to validate the membrane-feeding assay, sera collected from these same volunteers were subjected to a standard membrane-feeding assay. The data suggest that areas with intense but seasonal transmission might be feasible sites for testing transmission-blocking vaccines because of the high gametocytemic rates, high mosquito infectivity rates, and lack of pre-existing humoral-mediated transmission-blocking activity. The differences observed between field-based direct mosquito feeds and laboratory-based membrane feeding assays suggests that caution be used in interpreting Phase I study results in which laboratory-based membrane-feeding assays are used as a surrogate for vaccine efficacy. PMID- 9749650 TI - Heart autonomic innervation during the acute phase of experimental American trypanosomiasis in the dog. AB - Heart autonomic innervation was studied in dogs during the acute phase of the experimental infection with the Berenice-78 strain of Trypanosoma cruzi. A glyoxylic acid-induced fluorescence method for catecholamines and a thiocholine method for demonstrating acetylcholinesterase activity showed the sympathetic and the parasympathetic nerve fibers, respectively. At day 34 of infection, moderate to-intense rarefaction of both cholinergic and noradrenergic nerve fibers occurred in the atria of all animals coincident with moderate to intense myocarditis. In the ventricles, sympathetic denervation was clearly present only when the inflammatory processes were moderate to intense. Preliminary results on the chronic phase indicate that normal autonomic innervation coexists with an incipient chronic fibrosing myocarditis. PMID- 9749649 TI - Treatment of congenital Chagas' disease diagnosed and followed up by the polymerase chain reaction. AB - In 1991 and 1992, a prenatal screening of Trypanosoma cruzi infection was carried out using ELISA and indirect immunofluorescence techniques. A total of 840 blood samples from pregnant women, obtained at the Maternity Ward of the Hospital de Clinicas, National University of Asuncion (Asuncion, Paraguay), and 1,022 samples from the Regional Hospital of the San Pedro Department of Paraguay were examined. It was observed that 7.7% and 10.5%, respectively, of the pregnant women were serologically positive for infection with T. cruzi. When blood samples obtained from newborns on the day of birth or, at the most, on the first few days afterwards were examined by direct microscopic observation, an incidence of congenital transmission of 3% was found. These results are consistent with those of neighboring countries. When a serologic follow-up was conducted on the newborns until six months of age, the incidence of congenital transmission reached 10%. The same incidence rate was obtained when the samples collected during the first days after birth were examined by the polymerase chain reaction (PCR). Fifty-eight infants born to seropositive mothers were followed-up, two of which were positive by direct microscopic observation at birth, and four who were PCR-positive, but microscopy-negative at birth. None of the infants were positive for IgM at birth. The infected babies were treated with benznidazole and were followed-up by serology and PCR for four years. We conclude that the PCR has a clear advantage over conventional techniques for the early detection of congenital transmission of T. cruzi infection, and for monitoring infants undergoing chemotherapy. PMID- 9749651 TI - Nitric oxides in plasma, urine, and cerebrospinal fluid in patients with severe falciparum malaria. AB - It has been suggested that nitric oxide (NO) plays an important role in the pathogenesis of severe falciparum malaria. Since NO has a very short half-life, nitrate and nitrite (NOx) levels, stable metabolites of NO, are used as measures of NO production. We measured plasma NOx levels in 24 adults with severe falciparum malaria on the Thai-Burmese border. After correction for renal function, there was no correlation between plasma NOx levels, or the total amount of NOx excreted in the urine, and disease severity. Plasma NOx levels decreased after the first 48 hr in all patients (P = 0.007), suggesting decreased NO production. The NOx levels in cerebrospinal fluid (CSF) correlated well with plasma NOx levels, but these did not show a correlation with coma depth, and were not significantly different from those in a healthy control group. These findings do not support the hypothesis that excessive NO production contributes to the pathogenesis of severe falciparum malaria. However, local changes in NO production, e.g., in the central nervous system, might not be reflected in the total NOx production or NOx levels in the CSF. PMID- 9749652 TI - Spa therapy: panacea or placebo? PMID- 9749653 TI - Views from funding agencies. National Institute of Mental Health. PMID- 9749654 TI - Use of spa therapy to improve the quality of life of chronic low back pain patients. AB - OBJECTIVES: This study assessed the effectiveness of adding spa therapy to usual drug treatment in chronic low back pain patients. METHODS: A total of 224 patients were assigned randomly to either a treatment (n=128) or a control (n=96) group. Subjects in both groups received usual drug therapy. Those in the treatment group also underwent spa therapy in Vittel, France, for 6 days a week for 3 consecutive weeks. Effectiveness was assessed using a quality-of-life scale (the Duke Health Profile), clinical measures, and the Roland and Morris disability questionnaire. Groups were compared using an analysis of variance with repeated measures. RESULTS: At both 3 weeks and 3 months, patients in the treatment group exhibited significant improvement in measures of: physical and mental dimensions of quality of life, anxiety, depression, pain duration, pain intensity, and functional disability. CONCLUSION: This study suggests that spa therapy is an effective treatment for chronic low back pain patients. PMID- 9749655 TI - Variation in surgical rates: a population study. AB - OBJECTIVES: The objective of this study was to compare the use of certain surgical services by physicians to three other professional groups (architects, lawyers, and nurses) and to the general population. To meet this objective, variations in six surgical procedures were studied: appendectomy, tonsillectomy, herniorrhaphy, cholecystectomy, cesarean section, and hysterectomy. METHODS: This was a cross-sectional study in the city of Valencia (Spain) from September 1992 to September 1993. The surveys were carried out by mail questionnaire. Two thousand ninety-six subjects were recruited by simple random sampling for each of the groups. The questionnaire contained questions referring to the interviewee and to each family member (spouse or partner and up to a maximum of four children). The proportion of subjects who stated that they had undergone a particular type of surgery was compared among the professional groups. The population denominator was the whole Valencia population. RESULTS: The global response rate was 83.7%. With the exception of herniorrhaphy, the rates of surgical procedures obtained for each procedure for the different groups were not significantly different. The frequency of operations for each group differed according to sex and age. The risk of undergoing tonsillectomy was significantly higher among the general population when there were no medical friends or relatives than when there were. CONCLUSIONS: Barriers against access to surgical care did not seem to have played an important role in the use of surgical services. The frequency of operations for the best informed consumers with the most access to these services, that is, physicians, was very similar to that for other population groups. PMID- 9749657 TI - Trends and black/white differences in treatment for nonmetastatic prostate cancer. AB - OBJECTIVES: Controversy and uncertainty surround use of radical prostatectomy, radiation therapy, and conservative symptomatic management in treating elderly men with nonmetastatic prostate cancer. Prior studies have demonstrated variations in use of these therapies by patient age, race, and geographic region. This study examined trends in treatment for nonmetastatic prostate cancer in black and white men aged 65 and older during the period 1986 to 1993. The study also explored factors related to use of initial therapies in these men. METHODS: A cohort of 52,915 men (48,410 white; 4,505 black) obtained from the linked SEER Medicare dataset was used in an observational design. Various sociodemographic and clinical measures were incorporated in the analysis. RESULTS: For both races, use of aggressive therapy had increased with time, although this trend appears to be slowing. Black men were less likely to undergo radical prostatectomy than were white men, but use of radiation therapy did not differ markedly by race. High socioeconomic status and a lack of comorbid conditions were among the factors predictive of aggressive therapy receipt. The relation between race and receipt of aggressive therapy was dependent on whether prostate cancer was detected by transurethral resection of the prostate. Sociodemographic and clinical characteristics explained approximately half the difference between black men and white men in radical prostatectomy use. CONCLUSIONS: This study documents racial differences and changing practice patterns in the treatment of nonmetastatic prostate cancer in elderly men. Further research is required to more fully understand reasons for racial differences, as well as to promote rational use of health care resources. PMID- 9749656 TI - Agreement between administrative files and written medical records: a case of the Department of Veterans Affairs. AB - OBJECTIVES: This study examined the reliability of Department of Veterans Affairs' health information databases concerning patient demographics, use of care, and diagnoses. METHODS: The Department of Veterans Affairs' Patient Treatment files for Main, Bed-section (PTF) and Outpatient Care (OCF) were compared with medical charts and administrative records (MR) for a random national sample of 1,356 outpatient visits and 414 inpatient discharges to Department of Veterans Affairs' facilities between July 1 and September 30, 1995. Records were uniformly abstracted by a focus group of utilization review nurses and medical record coders blinded to administrative file entries. RESULTS: Reliability was adequate for demographics (kappa approximately 0.92), length of stay (agreement=98%), and selected diagnoses (kappa ranged 0.39 to 1.0). Reliability was generally inadequate to identify the treating bedsection or clinic (kappa approximately 0.5). Compared with medical charts, Patient Treatment Files/Outpatient Care Files reported an additional diagnosis per discharge and 0.8 clinic stops per outpatient visit, resulting in higher estimates of disease prevalence (+39% heart disease, +19% diabetes) and outpatient costs (+36% per unique outpatient per quarter). CONCLUSIONS: In the absence of pilot work validating key data elements, investigators are advised to construct health and utilization data from multiple sources. Further validation studies of administrative files should focus on the relation between process of data capture and data validity. PMID- 9749658 TI - Performance of a self-administered mailed version of the Quality of Well-Being (QWB-SA) questionnaire among older adults. AB - OBJECTIVES: The Quality of Well-Being questionnaire is a measure of health related quality of life (HRQoL) that has several desirable properties. Its widespread use has been hindered because it is difficult to administer. To overcome this limitation, a new self-administered form has recently been developed. This study examined the feasibility of using the Quality of Well-Being Self-Administered (QWB-SA) questionnaire in an older population. METHODS: The Quality of Well-Being-Self-Administered questionnaire was sent to 430 community dwelling individuals aged 65 years and older who were randomly selected from primary care physicians' offices. Response patterns, scaling distributions, and the acceptability of the survey were examined for all respondents. The results of the QWB-SA questionnaire were compared to the Sickness Impact Profile (SIP) and the Medical Outcomes Study 36-item Short-Form Health Survey (SF-36) for those individuals who also had completed the latter two surveys approximately 10 months earlier and whose health had not changed substantially in the meantime. RESULTS: Three hundred and one older adults (70%) responded. The mean QWB-SA questionnaire score was 0.7035. The scores were not skewed, and there were no floor or ceiling effects. The mean time to complete the QWB-SA questionnaire was 14.2 minutes, which was significantly shorter than for the SIP (19.3 minutes) but significantly longer than for the SF-36 (12.5 minutes). Subjects rated their satisfaction with the QWB-SA questionnaire somewhat lower than for the SIP and similar to SF-36. Correlations between the QWB-SA questionnaire and the SIP and SF-36 were moderate and were generally stronger for measures of physical health than for other domains such as mental health. CONCLUSIONS: The self-administered QWB questionnaire was acceptable to older respondents, and it correlated with other measures of health-related quality of life. It can be considered as a candidate for some research applications among older adults. PMID- 9749659 TI - Validation of the 36-item short-form Health Survey (Hebrew version) in the adult population of Israel. AB - OBJECTIVES: In the past few years, the SF-36 Health Survey has drawn considerable attention from researchers in non-English-speaking countries. This report contributes to the growing body of literature on this instrument by reporting the results of a national study conducted in Israel. The study examined the psychometric properties of the Hebrew translation based on a sample of the adult population of Israel and evaluated the results from a cross-national perspective. METHODS: The sample included 2,030 adults drawn from the Jewish population, aged 45 to 75 years. The SF-36 Health Survey was administered in face-to-face interviews as part of a broader health study. RESULTS: The pattern of correlations among items and the internal consistency scores pointed to high reliability. Confirmatory factor analysis using the Amos 3.61 program supported the hypothesized factorial structure. Specifically, the items clustered around eight health dimensions, as was found in studies in other societies. Clear and statistically significant differences in the SF-36 Health Survey scores were found among age groups and population groups distinguished by the degree of chronic health problems. CONCLUSIONS: Results of the analysis indicate that the instrument provided an appropriate measure of general health status. The findings clearly indicate that the translation into the Hebrew language and the application of the instrument to a culturally heterogeneous population did not diminish the qualities of the instrument. They also point to certain items that might be modified to reduce problems of synonimity and embeddedness. PMID- 9749660 TI - Predicting outcome in the intensive care unit using scoring systems: is new better? A comparison of SAPS and SAPS II in a cohort of 1,393 patients. GiViTi Investigators (Gruppo Italiano per la Valutazione degli interventi in Terapia Intensiva). Simplified Acute Physiology Score. AB - OBJECTIVES: This study sought to compare the performance of the old and new versions of the Simplified Acute Physiology Score, SAPS and SAPS II, in classifying patients according to the risk of hospital mortality. METHODS: To compare the performance of the two systems, measures of association between the scores and observed mortality were adopted, together with discrimination (area under the Receiver Operating Characteristics curve) and calibration (goodness-of fit statistics) estimates. Subjects were 1,393 eligible patients recruited during 1 month in 1994. The outcome measure was vital status at hospital discharge. RESULTS: SAPS II was associated more strongly with hospital mortality than the earlier version. SAPS II also had better discrimination ability than SAPS (area under Receiver Operating Characteristics curve 0.80 versus 0.74) and predicted an overall number of deaths (416.5) closer to the observed figure (475) than SAPS (267.7). Conversely, neither SAPS nor SAPS II fitted our data. Both P values derived from goodness-of-fit statistics were lower than 0.05. CONCLUSIONS: SAPS II offers a real improvement compared with SAPS in its ability to explain hospital mortality, but its standard parameters do not fit our data from Italy. The role and impact of potential determinants of this lack of fit, such as random errors and confounders related to casemix and/or quality of care should be clarified before this scoring system be used outside formal research projects. Special caution is suggested when SAPS II is adopted to predict mortality to compare intensive care unit performance across different countries and systems of care. PMID- 9749661 TI - A profile of health care utilization of the disabled population in Manitoba. AB - OBJECTIVES: This study profiled health care utilization by disabled and nondisabled individuals in the Canadian province of Manitoba to evaluate the association between health care utilization and disability. METHODS: Age standardized annualized utilization rates were calculated according to sex using longitudinal data on individual encounters with the Manitoba health care system from 1983 to 1990. Associations between severity of disability, number of prior chronic conditions, and prospective utilization were examined using multivariate regressions. RESULTS: Utilization patterns of the mildly disabled and the nondisabled differed only slightly. Severely disabled individuals had much higher rates of contact and consumed more resources, even after controlling for chronic conditions. The severely disabled accounted for 3% of the population and consumed 16% of hospital days and 7% of physician costs annually. CONCLUSIONS: The findings emphasize the importance of incorporating measures of disability in health services research. Both the severity of disability and the number of chronic conditions had independent value in predicting health care utilization. This has important implications for data collection and for the allocation of health care resources for research, which has traditionally been targeted toward fatal chronic conditions. PMID- 9749662 TI - What if socioeconomics made no difference?: access to a cadaver kidney transplant as an example. AB - OBJECTIVES: Several studies have noted the impact of socioeconomic factors on access to expensive medical care, but none of those studies controlled for self reported health and functional status or attitudes about treatment alternatives when analyses were completed. Because these factors may be correlated with socioeconomic status, the failure to control for them may have led to bias in other studies. The authors merged data from secondary sources with telephone survey data from a national sample of 456 end-stage renal disease patients to show how estimates of the effects of socioeconomic factors change when self reported health and functional status and attitudes about treatment are incorporated into statistical models. The authors also showed how kidney transplant rates would change if socioeconomic factors no longer influences organ allocation decisions. METHODS: Weibull proportional hazard analyses were used to show relationships between socioeconomic measures and waiting list entry and kidney transplant rates, before versus after accounting for self-reported health and functional status, attitudes about treatment, and other variables. Simulation analyses were used to estimate the number of waiting list spots and transplant operations that would move from economically advantaged to disadvantaged persons if socioeconomics no longer influenced organ allocation decisions. RESULTS: Incorporating information about health and functional status, attitudes about treatment, and other factors into the hazard models often reduced the estimated impact of socioeconomic measures on the odds of (1) being on a waiting list for a cadaver kidney transplant and (2) receiving a transplant. Simulations showed that 30 to 65 waiting list spots or transplant operations per 1,000 patients would shift from economically advantaged to disadvantaged persons if socioeconomics no longer influenced organ allocation decisions. CONCLUSIONS: Successful efforts to level the playing field would result in substantial redistributions of kidney transplants from economically advantaged to disadvantaged persons. PMID- 9749663 TI - Spanish language translation and initial validation of the functional assessment of cancer therapy quality-of-life instrument. AB - OBJECTIVES: There is a need in the United States for culture-equivalent assessment of health-related quality of life, particularly among people who speak different languages and among those with low literacy skills. This report summarizes the adaptation of the Functional Assessment of Cancer Therapy (FACT) Scales for use with Spanish-speaking cancer patients, including those with low literacy. METHODS: The Spanish language version of the general Functional Assessment of Cancer Therapy scale plus five disease-specific subscales (breast, lung, colorectal, head and neck, HIV infection) were translated, reviewed, and revised, then evaluated in interviews with Spanish-speaking patients from the mainland United States and Puerto Rico. An iterative forward-backward-forward sequence of item translation, expert bilingual/bicultural advisor review, pretesting interviews with 92 patients, and further expert advisory input were used to establish semantic, content, and partial technical equivalence. RESULTS: The Functional Assessment of Cancer Therapy-General and five disease-specific subscales were translated successfully into wording that was easily understood and answered, leading to psychometric and scoring data similar to that of the English version. All but one of the 28 Functional Assessment of Cancer Therapy General items and all of the disease-specific items were seen as culturally relevant. The result is a document that underwent iterative forward-backward translation and evaluation and was pretested successfully with native Spanish speaking oncology patients living in the Central United States and Puerto Rico. CONCLUSIONS: The Functional Assessment of Cancer Therapy-General and five disease specific subscales have been translated successfully into Spanish using a thorough translation and initial validation methodology. The methods and data provide a model for preparing a health status questionnaire for cross-cultural validation. The questionnaire is available for use in clinical trials and clinical practice. PMID- 9749664 TI - Is community-based treatment an add-on or a substitution for hospital treatment of alcoholism? Some evidence from Canada. AB - OBJECTIVES: This study determined whether the development of community treatment of alcohol problems acted as an add-on or a substitution for the utilization of inpatient hospital services in Ontario. METHODS: Complex modelling and graphic analyses using econometric multiple regression techniques were performed on data for the 48 counties of Ontario (Canada) for the period 1972 to 1988, combining both cross-sectional and time series analysis. RESULTS: After controlling for differences in alcohol consumption, in health care characteristics such as the supply of physicians or hospital occupancy rates, and in socioeconomic characteristics of the population, when community treatment became available, hospital utilization for the treatment of alcohol problems decreased and community services were substituting for hospital treatment. In addition, nonresidential services had an overall greater importance in producing this effect (elasticities at the mean of -0.11 to -0.14 depending on the region) than community-based residential treatment. The effect was larger in the southern than in the northern counties of Ontario. Testing of the modelling techniques showed statistically significant and satisfactory modelling of the forces at work. CONCLUSIONS: Where community-based treatment was available, it was used in preference to inpatient hospital treatment; however, there may be a slightly more complex relationship present in the southern urban counties (which contain the larger metropolitan areas) than in the northern and southern rural counties.. PMID- 9749665 TI - Physicians' perceptions of managed care: a structural equation model assessment of key dimensions. AB - OBJECTIVES: To assess the interrelationship of physicians' own interest with that of the methods and the anticipated benefits of managed care, the authors developed a new instrument to assess physician's perception of job satisfaction, risk, need to adapt their practice behavior, quality of care, cost of care, and access under managed care. METHODS: One hundred sixty-one attending physicians of an urban public hospital in a metropolitan area with low to moderate managed care penetration participated. A 24-item questionnaire with good psychometric properties was developed based on literature reviews, qualitative interviews with the key informants, and focus group discussion among a group of selected physician representatives. Confirmatory factory analysis and structural equation models were applied. RESULTS: The study reveals that when physicians perceived that high job satisfaction would ensue, they also perceived that quality and access to care would improve under managed care. Physician's perception of the need to modify their practice behavior was associated with a perception of increasing the cost of care. Risk sharing, from the physician's perspective, did not translate to cost savings as expected by managed care organizations, and only resulted in a fractional improvement on a perception of quality and access of care. CONCLUSIONS: Although this study reports the perceptions of a small group of physicians from a single hospital, the data suggest that increasing quality and decreasing cost may be included in the same equation, if physician job satisfaction also is included through organizational support and user-friendly work environment. PMID- 9749666 TI - Isolation, characterisation and mapping of simple sequence repeat loci in potato. AB - Solanum tuberosum L. DNA sequences containing simple sequence repeat (SSR) motifs were extracted from the EMBL database, cDNA and selectively enriched small-insert DNA libraries. Enrichment was achieved using either triplex affinity capture or single-strand hybridisation selection. One hundred and twelve primer pairs which successfully amplified products of the correct size from potato DNA were ultimately designed and synthesised. Ninety-eight of these revealed length polymorphisms in a panel of four diploid and two tetraploid clones, in agreement with the high information content of this class of markers which has been found in other species. All of the markers were assigned a quality score of 1-5 based on their potential usefulness. Eighty-nine loci from 65 of the primer pairs were located on two genetic linkage maps of potato by segregation analysis of the amplified alleles. Fifty-two of the SSRs were clearly single locus. The maps were aligned using 23 SSR primer pairs and 13 RFLP loci mapped in both populations. The markers described constitute a class which should replace Restriction Fragment Length Polymorphisms (RFLP) as the markers of choice for future genetic studies in potato. The sequences of the primers, together with other information on these markers are provided. PMID- 9749667 TI - Synthesis of glutamine, glycine and 10-formyl tetrahydrofolate is coregulated with purine biosynthesis in Saccharomyces cerevisiae. AB - Glutamine, glycine and 10-formyl tetrahydrofolate are consumed during de novo purine biosynthesis. We have found that, in Saccharomyces cerevisiae, synthesis of these cosubstrates is coregulated with synthesis of enzymes of the purine biosynthetic pathway. Analysis of three genes required for synthesis of glutamine, glycine and 10-formyl tetrahydrofolate (GLN1, SHM2 and MTD1, respectively) shows that their expression is repressed by adenine and requires the transcription factors Baslp and Bas2p. Northern analysis reveals that regulation of SHM2 and MTD1 expression by adenine takes place at the transcriptional level. We also show that Bas1p and Bas2p bind in vitro to the promoters of the SHM2 and MTD1 genes, and that mutations in the consensus Bas1p binding sequences strongly affect expression of these genes in vivo. Finally, we have found that a SHM2-lacZ fusion is expressed at a significantly higher level in a bas2-2 disrupted strain than in bas1-2 or bas1-2 bas2-2 mutant strains. The BAS1-dependent, BAS2-independent expression of SHM2-lacZ suggests that, in the absence of Bas2p, Bas1p can interact with another protein partner to activate SHM2 expression. PMID- 9749668 TI - Structure of the Chlorella Zepp retrotransposon: nested Zepp clusters in the genome. AB - Zepp elements found in the telomeric region of Chlorella chromosomes show the characteristic features of non-viral (LINE-like) retrotransposons, including a poly(A) tail, 5' truncations, a retroviral reverse transcriptase-like ORF and flanking target duplications. We have isolated and characterized a full-length Zepp element (8943 bp long) from Chlorella chromosome V. Some peculiar features of this element, including nested integration, two ORF structures, a long 3' noncoding region and a possible promoter region are compared with those of the Drosophila telomeric retrotransposons HeT-A and TART. The Chlorella chromosome Zepp system appears to represent an intermediate stage between canonical telomerase-telomeres and Drosophila retrotransposon-telomeres. PMID- 9749669 TI - Mutation of the gene for the second-largest subunit of RNA polymerase I prolongs the period length of the circadian conidiation rhythm in Neurospora crassa. AB - The period length of the circadian conidiation rhythm was examined in a mutant strain of Neurospora crassa, un-18, that is temperature sensitive for mycelial growth. The un-18 mutant showed a temperature-sensitive phenotype with respect to both mycelial growth and the period length of the conidiation rhythm. Below 22 degrees C, the un-18 mutation did not affect the period length, but at temperatures between 22 degrees C and 32 degrees C, the period length of the un 18 mutant was approximately 2 h longer than that of the wild-type strain. The un 18+ gene was cloned and was found to encode the second-largest subunit of RNA polymerase I, which is involved in the synthesis of rRNA. These results indicate that a defect in ribosome synthesis, which must result in a lower rate of protein synthesis, lengthens the period of the circadian conidiation rhythm in Neurospora. PMID- 9749670 TI - Single mating type-specific genes and their 3' UTRs control mating and fertility in Cochliobolus heterostrophus. AB - To determine the number of proteins required for mating type (MAT) locus regulated control of mating in Cochliobolus heterostrophus, MAT fragments of various sizes were expressed in MAT deletion strains. As little as 1.5 kb of MAT sequence, encoding a single unique protein in each mating type (MAT-1 and MAT-2), conferred mating ability, although an additional 160 bp of 3' UTR was needed for production of ascospores. No other mating type-specific genes involved in mating identity or fertility were found. Thus, although homologs of the C. heterostrophus MAT-1 and MAT-2 genes exist in the filamentous ascomycetes Neurospora crassa and Podospora anserina, C. heterostrophus does not appear to have mating type-specific homologs of two additional genes required by both N. crassa and P. anserina for successful sexual reproduction. Three genes were identified in the common DNA flanking the MAT locus: a gene encoding a GTPase activating protein and an ORF of unknown function lie 5' while a beta-glucosidase encoding gene lies found 3'. None of these genes appears to be involving in the mating process. PMID- 9749671 TI - mRNA translation in yeast during entry into stationary phase. AB - The expression of some Saccharomyces cerevisiae genes is induced as cells enter stationary phase. Their mRNAs are translated during a period in the growth cycle when the translational apparatus is relatively inert, thereby raising the possibility that these mRNAs compete effectively for a limiting pool of translation factors. To test this idea, the translation of mRNAs carrying different 5'-leaders was compared during exponential growth and after entry into stationary phase upon glucose starvation. Closely related sets of lacZ mRNAs, carrying 5'-leaders from the PYK1, PGK1, RpL3, Rp29, HSP12, HSP26 or THI4 mRNAs, were studied. These mRNAs displayed differing translational efficiencies during exponential growth, but their relative translatabilities were not significantly affected by entry into stationary phase, indicating that they compete just as effectively under these conditions. Polysome analysis revealed that the wild-type PYK1, ACT1 and HSP26 mRNAs are all translated efficiently during stationary phase, when the translational apparatus is relatively inert. Also, significant levels of the translation initiation factors eIF-2alpha, eIF-4E and eIF-4A were maintained during the growth cycle. These data are consistent with the idea that, while translational activity decreases dramatically during entry into stationary phase, yeast cells maintain excess translational capacity under these conditions. PMID- 9749672 TI - Suppression of a +1 T mutation by a nearby substitution in the mitochondrial cox1 gene of Chlamydomonas reinhardtii: a new type of frameshift suppression in an organelle genome. AB - In Chlamydomonas reinhardtii, mutants defective in the cytochrome pathway of respiration lack the capacity to grow under heterotrophic conditions (in darkness on acetate). In the dark- strain duM18, a + 1 T addition in a run of four Ts, located at codon 145 of the mitochondrial cox1 gene encoding subunit I of cytochrome c oxidase, is responsible for the mutant phenotype. A leaky revertant (su11) that grows heterotrophically at a lower rate than wild-type cells was isolated from dum18. Its respiration sensitivity to cyanide was low and its cytochrome c oxidase activity was only 4% of that of the wild-type enzyme. Meiotic progeny obtained from crosses between revertant and wild-type cells inherited the phenotype of the mt- parent, showing that the suppressor mutation, like dum18 itself, is located in the mitochondrial genome. In order to map the su11 mutation relative to dum18, a recombinational analysis was performed on the diploid progeny. It demonstrated that su11 was very closely linked to the dum18 mutation less than 20-30 bp away. The cox1 gene of the su11 revertant was then sequenced. In addition to the + 1 T frameshift mutation still present at codon 145, an A-->C substitution was found at codon 146, leading to the replacement of a glutamic acid by an alanine in the polypeptide chain. No other mutations were detected in the cox1 coding sequence. As the new GCG codon (Ala) created at position 146 is very seldom used in the mitochondrial genome of C. reinhardtii, we suggest that the partial frameshift suppression by the nearby substitution is due to an occasional abnormal translocation of the ribosome (+ 1 base shift) facilitated both by the run of Ts and the low level of weak interaction of alanyl tRNA. PMID- 9749673 TI - Analysis of a Streptomyces antibioticus chromosomal region involved in oleandomycin biosynthesis, which encodes two glycosyltransferases responsible for glycosylation of the macrolactone ring. AB - A 6-kb region from the chromosome of Streptomyces antibioticus, an oleandomycin producer, was cloned and sequenced. This region was located between the 3' end of the gene encoding the third subunit of the oleandomycin type I polyketide synthase and the oleP and oleB genes, which encode a cytochrome P450 monooxygenase and an oleandomycin resistance gene, respectively. Analysis of the nucleotide sequence revealed the presence of five genes encoding a cytochrome P450-like protein (oleP1), two glycosyltransferases (oleG1 and oleG2) involved in the transfer of the two 6-deoxysugars (L-oleandrose and D-desosamine) to the oleandomycin macrolactone ring, a methyltransferase (oleM1), and a gene (oleY) of unknown function. Insertional inactivation of this region by gene disruption generated an oleandomycin non-producing mutant which accumulated a compound that, according to mass spectrometry analysis, could correspond to the oleandomycin macrolactone ring (oleandolide), suggesting that the mutation affects oleandrosyl glycosyltransferase. PMID- 9749674 TI - Downstream elements from the pea albumin 1 gene confer sulfur responsiveness on a reporter gene. AB - The levels of mRNAs for some of the sulfur-rich proteins in seeds are regulated by the level of sulfur supplied to the plants. In peas, there is a mechanism that lowers the level of mRNA for legumin and pea albumin 1 (PA1) when plants are grown under sulfur-deficient conditions. This mechanism acts after transcription initiation. In this study, a gene encoding PA1 was expressed in leaves of transgenic tobacco. Expression of the gene was controlled by the level of sulfur supplied to the plants, mimicking the behaviour of the intact gene in peas. A gene encoding a different high-sulfur protein, ovalbumin, was unresponsive to sulfur status and was used as a reporter gene to test defined regions of the PA1 gene for sulfur responsiveness. These constructs, together with a set of PA1 gene deletions, were tested in transgenic tobacco and yielded the following observations: the PA1 gene was sensitive to sulfur status in the leaf as well as the seed; intron processing of the PA1 transcript was not required for sensitivity to sulfur stress; both the coding region and the 3' flanking regions of the PA1 gene contained sequences which conferred sensitivity to sulfur stress; the sulfur-responsive sequence in the 3' region was contained within a 134 nucleotide segment downstream of the end of the coding sequence. We conclude that there are at least two downstream elements which confer sensitivity to sulfur supply. PMID- 9749675 TI - CRP down-regulates adenylate cyclase activity by reducing the level of phosphorylated IIA(Glc), the glucose-specific phosphotransferase protein, in Escherichia coli. AB - The cellular cAMP level is markedly down-regulated by cAMP receptor protein (CRP) in Escherichia coli. CRP regulates adenylate cyclase both at the level of transcription of its structural gene cya and at the level of enzyme activity. We established a method to determine the phosphorylation state of IIA(Glc), the glucose-specific phosphotransferase protein, in intact cells. We found that IIA(Glc) exists predominantly in the unphosphorylated form in wild-type cells growing in LB medium, while it is largely phosphorylated in crp or cya cells. Disruption of the ptsG gene that codes for the membrane component of the major glucose transporter (IICB(Glc)), and/or the fruF gene coding for FPr (fructose specific hybrid phosphotransferase protein), did not affect the phosphorylation state of IIA(Glc). When IICB(Glc) was overproduced in the presence of glucose, the levels of both cAMP and phosphorylated IIA(Glc) in crp cells were concomitantly decreased to wild-type levels. In addition, when His-90 in IIA(Glc) was replaced by glutamine, both phosphorylation of IIA(Glc) and the overproduction of cAMP in crp cells were eliminated. We also found that extracts of crp+ cells markedly stimulate dephosphorylation of IIA(Glc)-P in vitro. We conclude that CRP-cAMP down-regulates adenylate cyclase primarily by reducing the level of phosphorylated IIA(Glc). The data suggest that unspecified proteins whose expression is under the control of CRP-cAMP are responsible for this regulation. PMID- 9749676 TI - Insertional mutagenesis of Aspergillus fumigatus. AB - We have investigated transformation with heterologous DNA as a method for insertional mutagenesis of Aspergillus fumigatus. Two methods, polyethylene glycol-mediated transformation of protoplasts and electroporation of germinating spores, were used to establish conditions leading to single-copy integration of transforming DNA at different genomic sites. We have assessed the effect of restriction enzyme-mediated integration (REMI) for both methods. Non-REMI protoplast transformation led to integration of multiple copies of transforming DNA in the majority of transformants. Results of REMI with protoplast transformation varied depending on the enzyme used. Low concentrations of several restriction enzymes stimulated transformation, but of ten enzymes investigated only REMI with XhoI and KpnI resulted in single-copy integration of transforming DNA for the majority of transformants. For protoplast transformation with XhoI- or KpnI-based REMI, 50% and 76% of insertions, respectively, were due to integrations at a genomic enzyme site corresponding to the enzyme used for REMI. Electroporation of spores without addition of restriction enzyme resulted in a high transformation efficiency, with up to 67% of transformants containing a single copy of transforming DNA. In contrast to protoplast transformation, electroporation of spores in the presence of a restriction enzyme did not improve transformation efficiency or lead to insertion at genomic restriction sites. Southern analysis indicated that for both protoplast transformation with REMI using KpnI or XhoI and for electroporation of spores without addition of restriction enzymes, transforming DNA inserted at different genomic sites in a high proportion of transformants. PMID- 9749677 TI - Orientation-dependent enhancement by H-NS of the activity of the type 1 fimbrial phase switch promoter in Escherichia coli. AB - Phase variation of type 1 fimbriation in Escherichia coli is associated with the inversion of a 314-bp DNA element, positioned proximal to and upstream of fimA, which encodes the major type 1 fimbrial subunit. This DNA switch region contains a promoter that drives transcription of fimA only when the switch is in the ON orientation. Using chromosomal and plasmid-borne lacZ reporter cassettes, we show here how the global regulator H-NS affects the activity of the fimA promoter. In phase-locked reporter cassettes the activity of the fimA promoter was found to be enhanced in a hns-positive background, but only when the switch was in the ON orientation. Also, the number of fimbriae produced by a phase-locked ON strain was significantly higher in a hns-positive background. By means of competitive gel retardation and DNase I protection assays, H-NS binding to DNA segments adjacent to and within the phase switch region was demonstrated. PMID- 9749678 TI - Answering questions with an electroencephalogram-based brain-computer interface. AB - OBJECTIVE: To demonstrate that humans can learn to control selected electroencephalographic components and use that control to answer simple questions. METHODS: Four adults (one with amyotrophic lateral sclerosis) learned to use electroencephalogram (EEG) mu rhythm (8 to 12Hz) or beta rhythm (18 to 25Hz) activity over sensorimotor cortex to control vertical cursor movement to targets at the top or bottom edge of a video screen. In subsequent sessions, the targets were replaced with the words YES and NO, and individuals used the cursor to answer spoken YES/NO questions from single- or multiple-topic question sets. They confirmed their answers through the response verification (RV) procedure, in which the word positions were switched and the question was answered again. RESULTS: For 5 consecutive sessions after initial question training, individuals were asked an average of 4.0 to 4.6 questions per minute; 64% to 87% of their answers were confirmed by the RV procedure and 93% to 99% of these answers were correct. Performances for single- and multiple-topic question sets did not differ significantly. CONCLUSIONS: The results indicate that (1) EEG-based cursor control can be used to answer simple questions with a high degree of accuracy, (2) attention to auditory queries and formulation of answers does not interfere with EEG-based cursor control, (3) question complexity (at least as represented by single versus multiple-topic question sets) does not noticeably affect performance, and (4) the RV procedure improves accuracy as expected. Several options for increasing the speed of communication appear promising. An EEG-based brain-computer interface could provide a new communication and control modality for people with severe motor disabilities. PMID- 9749679 TI - Topical morphine in a canine model: a pilot study. AB - OBJECTIVE: To determine if topical morphine can enter the synovial cavity and the effect of ultrasound on this process. DESIGN: A randomized control trial to investigate which body fluids morphine enters after topical application. SETTING: A university animal laboratory. SUBJECTS: Ten mongrel dogs raised by the Comparative Medicine Department. All animals were certified to be free of disease, all had received standard scheduled immunizations, and none had been used for any other research. INTERVENTION: Topical morphine and ultrasound or topical morphine and sham ultrasound was applied to the knees of the dogs. Samples were obtained afterward from synovial fluid, serum, and urine, and were analyzed for the presence of morphine. MAIN OUTCOME MEASURES: Blood samples were collected every 60 minutes for 240 minutes, urine samples were collected at 120 minutes and 240 minutes, and synovial joint fluid was collected at 120 minutes and 240 minutes. The process of collection and analysis was the same for dogs treated with topical morphine and ultrasound and those treated with topical morphine and sham ultrasound. Fisher's exact test was used to test for an association between the use of ultrasound and the presence of morphine in the synovial fluid, serum, or urine. Two-sample t tests were used to test for group differences in mean body weight. RESULTS: All samples (synovial fluid, serum, and urine) were negative at time zero. All of the subsequent serum samples were negative for morphine. Two or three of the dogs in each group of five (ultrasound or sham ultrasound) had positive urine and synovial fluid samples at 120 and 240 minutes. Ultrasound did not affect the results. Body weight of the dogs influenced the results, with lighter animals having a significantly larger percentage (p=.03) of synovial fluid samples positive for morphine. CONCLUSION: Ultrasound did not affect the absorption of topical morphine in this canine model. Body weight may have influenced the results. Dogs that tested positive for morphine in synovial fluid had a lower mean body weight than dogs that did not test positive (p=.03). PMID- 9749680 TI - ABILHAND: a Rasch-built measure of manual ability. AB - OBJECTIVE: To apply the Rasch measurement model to the development of a clinical tool for measuring manual (dis)ability (ABILHAND). DESIGN: Manual ability was evaluated in terms of the difficulty perceived by a hand-impaired patient on 57 representative unimanual or bimanual activities. SETTING: A clinical laboratory. PATIENTS: Eighteen rheumatoid arthritis patients (14 women, 4 men) were interviewed after wrist arthrodesis (10 right, 4 left, and 4 both wrists). Their ages ranged from 38 to 77 years, time since diagnosis ranged from 7 to 41 years, and time since surgery ranged from 0.5 to 17 years. MAIN OUTCOME MEASURE: ABILHAND, administered at a mean duration of 7 years after arthrodesis. RESULTS: Forty-six of the 57 items define a common, single manual ability continuum with widespread measurement range and regular item distribution. Items relating to feeding, grooming, and dressing upper body worked consistently with their counterparts in other disability scales. More difficult items extend the measurement range beyond that of most existing manual ability scales. CONCLUSION: Even in a small sample of patients, using the Rasch methodology enabled the investigators to produce a useful scale of manual (dis)ability and to define manual ability as a unique construct, at least in patients with rheumatoid arthritis. PMID- 9749681 TI - The sit-to-stand movement in stroke patients and its correlation with falling. AB - OBJECTIVE: To use kinetic assessment of the sit-to-stand movement as a means of sorting out those stroke patients at risk for falling. DESIGN: A retrospective study, using a force platform to assess sit-to-stand performance and to determine its correlation with falls in stroke patients. SETTING: Hospital-based rehabilitation units. METHODS: Thirty-three stroke patients (18 fallers, 15 nonfallers) and 25 age-matched healthy subjects were included in this study. Subjects sat in an adjustable chair with their feet on two force plates and performed the standing up/sitting down movement at a self-paced, comfortable speed. RESULTS: The rate of rise in force (dF/dT) was significantly lower in stroke fallers than in stroke nonfallers and healthy subjects (23.78+/-17.38, 55.23+/-31.24, and 85.96+/-42.4 percent body weight per second, respectively [p < .005]). The center of pressure sway in mediolateral direction during rising/ sitting down was much greater in stroke fallers than in stroke nonfallers or healthy subjects (p < .05). Body weight distribution was asymmetric on the feet of stroke patients, with much more body weight on their sound side. CONCLUSIONS: The significantly lower rate of rise in force and greater postural sway while rising/sitting down may be useful in identifying stroke patients who are at risk for falling. PMID- 9749682 TI - Methylphenidate in early poststroke recovery: a double-blind, placebo-controlled study. AB - OBJECTIVE: To determine the efficacy and safety of methylphenidate in acute stroke rehabilitation. DESIGN: A prospective, randomized, double-blind, placebo controlled study. PATIENTS AND SETTING: Twenty-one stroke patients consecutively admitted to a community-based rehabilitation unit. INTERVENTION: Three-week treatment of methylphenidate (or placebo) in conjunction with physical therapy. Methylphenidate was started at 5mg and increased gradually to 30mg (15mg at 8:00AM and 15mg at 12:00 noon), and discontinued before discharge. MAIN OUTCOME MEASURES: Mood measures included the Hamilton Depression Rating Scale (HAM-D) and Zung Self-Rating Depression Scale (ZDS). Cognitive status was evaluated using the Mini-Mental State Exam (MMSE). Motor functioning was assessed using the Fugl Meyer Scale (FMS) and a modified version of the Functional Independence Measure (M-FIM). All measures were administered pretreatment and weekly thereafter. Side effects were measured after each increase in dosage and weekly. RESULTS: Patients receiving methylphenidate treatment scored lower on the HAM-D (F(1,18)=5.714, p=.028), lower on the ZDS (F(1,18)=4.206, p=.055), higher on the M-FIM (F(1,18)=5.374, p=.032), and higher on the FMS (F(1,9)=4.060, p=.075) than patients receiving placebo. CONCLUSION: Methylphenidate appears to be a safe and effective intervention in early poststroke rehabilitation that may expedite recovery. PMID- 9749683 TI - Transcutaneous electrical nerve stimulation: effect on peripheral nerve conduction, mechanical pain threshold, and tactile threshold in humans. AB - OBJECTIVES: To investigate the effect of different transcutaneous electrical nerve stimulation (TENS) parameters on nerve conduction in the human superficial radial nerve and on peripheral mechanical pain threshold (MPT) and tactile threshold (TT), and to further the current knowledge of the neurophysiologic effects of TENS. STUDY DESIGN: Fifty healthy human subjects were randomly allocated in equal numbers to a control group or one of four TENS groups to receive electrical stimulation consisting of four combinations of TENS pulse durations (50microsec and 200microsec) and frequencies (4Hz and 110Hz). In the TENS groups, TENS was applied under double-blind conditions for 15 minutes over the superficial radial nerve in the dominant forearm. Over a 1-hour period, compound action potentials, MPT readings, and TT readings were recorded bilaterally. RESULTS: Only one combination of TENS parameters (110Hz, 200microsec) effected consistent changes in all of the variables assessed, ie, TENS produced a significant increase in negative peak latency while simultaneously increasing both MPT and TT. CONCLUSION: The findings from this study suggest that at least part of TENS-mediated hypoalgesia is a consequence of a direct peripheral effect of TENS, although a "central" effect may not be excluded. PMID- 9749684 TI - Anodal block in F-wave studies. AB - OBJECTIVE: To determine whether F-wave results differ with the anode proximal or distal to the cathode, ie, if clinical anodal block exists. DESIGN: Prospective study of 30 healthy volunteers undergoing nerve conduction and F-wave studies in one median nerve. A needle cathode electrode was used with a surface anode placed alternately proximal and distal to the cathode. The same electromyographer performed all studies with a Dantec Counterpoint machine. RESULTS: F-wave latencies were essentially unaffected by distal versus proximal positioning. Minimum, maximum, and mean F-wave latencies correlated extremely highly (r=.973 to .988). For both F-wave and M-response latencies and amplitudes, differences between mean values obtained using the two methods were extremely small and were neither clinically nor statistically significant. The frequency of elicitation of F-waves may (p < .05) have been slightly (3.5%) lower when the anode was in the distal position. CONCLUSION: Anodal block is not seen in F-wave studies when using needle electrode stimulations. Reversing the stimulator does not seem to be required. Further study with surface stimulating electrodes is underway to confirm results. PMID- 9749685 TI - Electrophysiologic evaluation of denervated muscles in incomplete paraplegia using macro electromyography. AB - OBJECTIVE: To evaluate denervated muscles in persons with incomplete paraplegia due to thoracolumbar spinal injury (TLSI) using macro electromyography in determining indications for functional electrical stimulation (FES). DESIGN: A randomized clinical trial and a criterion standard. SETTING: A department of orthopedic surgery in a university hospital. PATIENTS AND OTHER PARTICIPANTS: Eighteen patients with incomplete paraplegia, including 11 with TSLI, and 50 healthy adults. INTERVENTION: Area and amplitude of macro motor unit potential (macro MUP) were measured at the tibialis anterior, the vastus lateralis, and the vastus medialis. The normal limits of macro MUP parameters were defined based on values from healthy subjects. Abnormal denervated muscles were detected by macro EMG and conventional EMG in paralytic patients. The correlation between macro MUP parameter values and muscle forces of the tibialis anterior and quadriceps femoris induced by electrical stimulation was analyzed. MAIN OUTCOME MEASURES: The number of abnormal muscles, parameter values, and muscle force induced by electrical stimulation. RESULTS: Abnormal muscles were found only in the TLSI patients and 13 abnormal muscles were detected by macro EMG only. The abnormal muscles defined by macro EMG showed insufficient contraction induced by electrical stimulation. The increase of parameter value negatively correlated with the muscle force (tibialis anterior area r=-.797, amplitude r=-.866; quadriceps area r=-.866, amplitude r=-.893; p < .001). CONCLUSIONS: These results suggest that macro EMG is useful in detecting denervated muscles, in determining indications for FES, and in predicting FES effects before implantation of electrodes. PMID- 9749687 TI - Rater reliability of Fahn's tremor rating scale in patients with multiple sclerosis. AB - OBJECTIVE: Assessment of movement disorders in patients with multiple sclerosis (MS) is difficult because of the complex nature of the movement disorders. The aim of this study was to determine the reliability of Fahn's Tremor Rating Scale (FTRS) in assessing movement disorders in patients with MS. METHOD: Videos were made of 10 patients with MS showing their rest, postural, action/intention, and goal-related movement disorders as well as their performance of spirometry, a volumetric task, and timed functional tasks. Ratings of tremor were carried out by one rater on two occasions 3 months apart and by 8 raters on one occasion using FTRS. RESULTS: Intrarater reliability was generally very good, with no significant "drift" in ratings over time. Interrater reliability was generally good, with some variation in interpretation of scoring criteria that may reflect raters' backgrounds. CONCLUSION: The FTRS is a reliable and potentially useful tool with which to assess movement disorders in patients with MS. PMID- 9749686 TI - Age, sex, and body mass index as determinants of back and hip extensor fatigue in the isometric Sorensen back endurance test. AB - OBJECTIVE: To study the ability of a widely used isometric back endurance test to measure lumbar back erector muscle fatigue and to assess the influence of age, sex, and body mass index (BMI) on back and hip extensor muscle fatigability (EMG spectral indices). DESIGN: Cross-sectional study of men and women without back problems. SETTING: Occupational health center and rehabilitation clinic in Finland. SUBJECTS: Experiment 1 consisted of 233 consecutive occupational health center customers (133 women, 100 men) without back problems. Experiment 2 consisted of 20 healthy women. INTERVENTION: Subjects performed the isometric Sorensen back endurance test up to 240sec in experiment 1 and to the limit of endurance in experiment 2. OUTCOME MEASURES: Raw surface EMG was recorded bilaterally over the belly of lumbar erector spinae muscles at L1-L2 and L4-L5 levels in experiment 1, and bilaterally over the medial paraspinal muscles at L1 L2, L3-L4, and L5-S1 levels and over the major hip extensor muscles (gluteus maximus and biceps femoris) in experiment 2. In both experiments, time to endurance was recorded (in experiment 1 up to 240sec). The EMG spectral median frequency (MF) decrease over time was used for the assessment of back and hip extensor fatigability. RESULTS: In experiment 1, the rate of change in paraspinal MF was greater in men than in women, indicating greater paraspinal fatigability in men. Multiple regression analysis indicated that the rate of MF decrease (fatigue) during the test was dependent on age and BMI in both sexes and that the effects of age and BMI were more pronounced in women than in men. Correlation analysis revealed that the rate of paraspinal muscle MF decrease was associated with endurance time and BMI in women and with endurance time and age in men. In experiment 2, the paraspinal muscles, as well as the hip extensor muscles, biceps femoris, and gluteus maximus, showed clear decreases in MF during the isometric endurance test in women. MF decrease was highly related to endurance time and BMI in women. CONCLUSIONS: Lumbar paraspinal muscle fatigability during the Sorensen test is influenced by subject characteristics. Further, the hip extensor muscles also significantly fatigue, indicating load sharing between back and hip extensor muscles during the test. According to these results, the validity of this widely used back endurance test in specifically measuring lumbar paraspinal muscle endurance is questionable, as is the direct comparison of test results between women and men. PMID- 9749688 TI - Magnetic resonance imaging of nonhealing pressure ulcers and myocutaneous flaps. AB - OBJECTIVE: To evaluate the use of magnetic resonance imaging (MRI) in making clinical decisions when assessing nonhealing pressure ulcers and nonhealing myocutaneous flaps for the presence of an abscess, osteomyelitis, sinus tracts, and fluid collections. DESIGN: Retrospective review of patient charts and radiographic studies. SETTING: Regional spinal cord injury center. SUBJECTS: Twelve patients who had MRI as part of their evaluation for a nonhealing pressure ulcer or myocutaneous flap. RESULTS: Seven patients had MRI for preoperative evaluation, four with a previous flap that had recurrent breakdown and three with a new grade III or IV ulcer. Five patients had MRI for postoperative evaluation of myocutaneous flaps with delayed healing. MRI was useful in identifying osteomyelitis in three patients and sinus tracts that required surgical revision in six patients. MRI was also used in two patients to assess the size of fluid collections postoperatively in determining whether the patients should be mobilized after surgery. These chronic nonhealing wounds resulted in multiple admissions and lengthy hospital stays and required multiple surgical revisions. Patients who did poorly with healing or had repeated breakdown tended to have concurrent issues such as poor self care, increased age, increased time of spinal cord injury, poor nutrition, or other medical problems. CONCLUSION: Chronic nonhealing pressure ulcers and myocutaneous flaps can be difficult to treat and evaluate with conventional methods. There are multiple reasons for failure to heal. MRI can be a useful tool for identifying some of these factors including osteomyelitis, fluid collections, abcesses, and sinus tracts in the perioperative period. Identifying the appropriate patient populations and clinical indications for the optimal use of MRI should be subject of further study. PMID- 9749689 TI - Cervicocephalic kinesthetic sensibility, active range of cervical motion, and oculomotor function in patients with whiplash injury. AB - OBJECTIVE: To investigate cervicocephalic kinesthetic sensibility, active range of cervical motion, and oculomotor function in patients with whiplash injury. DESIGN: A 2-year review of consecutive patients admitted to the emergency unit after whiplash injury. SETTING: An otorhinolaryngology department. PATIENTS AND SUBJECTS: Twenty-seven consecutive patients with diagnosed whiplash injury (14 men and 13 women, mean age, 33.8yrs [range, 18 to 66yrs]). The controls were healthy subjects without a history of whiplash injury. MAIN OUTCOME MEASURES: Oculomotor function was tested at 2 months and at 2 years after whiplash injury. The ability to appreciate both movement and head position was studied. Active range of cervical motion was measured. Subjective intensity of neck pain and major medical symptoms were recorded. RESULTS: Active head repositioning was significantly less precise in the whiplash subjects than in the control group. Failures in oculomotor functions were observed in 62% of subjects. Significant correlations occurred between smooth pursuit tests and active cervical range of motion. Correlations also were established between the oculomotor test and the kinesthetic sensibility test. CONCLUSION: The results suggest that restricted cervical movements and changes in the quality of proprioceptive information from the cervical spine region affect voluntary eye movements. A flexion/extension injury to the neck may result in dysfunction of the proprioceptive system. Oculomotor dysfunction after neck trauma might be related to cervical afferent input disturbances. PMID- 9749690 TI - Long-term survival of children and adolescents after traumatic brain injury. AB - OBJECTIVE: To obtain information on long-term mortality risk and life expectancy after traumatic brain injury (TBI), to improve planning and for counseling patients and their families. In contrast to the literature for spinal cord injury and other disabilities, there have been few such reports for TBI. DESIGN: Records were reviewed on 946 persons aged 5 to 21 years who had sustained TBI. All were patients who subsequently received disability services in California, 1987 to 1995. RESULTS: The chief predictors of mortality were basic functional skills such as mobility and self-feeding. After the initial high-risk period, mortality risk for TBI was much lower than for similarly functioning persons with cerebral palsy (a comparison group), although after 10 years the two sets of mortality rates had largely converged. For high-functioning persons, life expectancies were only 3 to 5 years shorter than for the general population. By contrast, the remaining life expectancy for those without mobility 6 months after injury was only 15 years. PMID- 9749691 TI - Comparison of the mechanical properties of the heel pad between young and elderly adults. AB - OBJECTIVE: To compare the mechanical properties of the human heel pad between young and aged adults. DESIGN: A 7.5-MHz linear-array ultrasound transducer was incorporated into a specially designed device to measure the thickness of the heel pad under different loads. The heel pad was compressed with serial increments of 0.5kg to a maximum of 3kg and then relaxed sequentially. Then the load-displacement curve of the heel pad during a loading-unloading cycle was plotted. PARTICIPANTS: Convenience sample of 33 volunteers without heel problems, aged 18 to 78 years, were divided into young (less than 40 years) and elderly (older than 60 years) groups. MAIN OUTCOME MEASURES: Unloaded heel-pad thickness, compressibility index, stiffness, and energy dissipation ratio were calculated from the load-displacement curves. Student's t-test was used to compare the mechanical properties of the heel between these two groups. RESULTS: The average unloaded heel-pad thickness was 1.76+/-.20cm in the young group and 2.01+/-.24cm in the elderly group (p < .001). The average compressibility index was 53.3%+/ 7.7% in the young group and 61.3%+/-5.5% in the elderly group (p < .001). Energy dissipation ratio representing shock absorbency of the heel pad, was 23.7%+/-6.9% in the young group and 35.3%+/-10% in the elderly group (p < .001). CONCLUSION: Unloaded heel-pad thickness, compressibility index, and energy dissipation ratio of the heel pad were significantly increased in the elderly group, indicating loss of the elasticity of the heel pad. The loss of elasticity may be responsible for the higher incidence of heel injury in elderly individuals. PMID- 9749693 TI - Alcohol use and readiness to change after spinal cord injury. AB - OBJECTIVE: To describe alcohol use and motivation to change drinking among persons with recent spinal cord injury (SCI). DESIGN: Survey SETTING: Acute inpatient rehabilitation program PATIENTS: Subjects were 58 patients with recent SCI assessed during inpatient rehabilitation. MAIN OUTCOME MEASURES: Short Michigan Alcoholism Screening Test (SMAST), Readiness to Change questionnaire (RTC), and alcohol use questions. RESULTS: Subjects were on average 39 years old, 88% were male, and 86% were Caucasian. Thirty-five percent of the total sample scored in the "alcoholic" range on the SMAST. Twenty-nine (50%) of the sample were considered "at-risk" drinkers. Of these, 6 (21%) were in the precontemplation phase, 13 (45%) were in the contemplation phase, and 10 (34%) were in the action phase with respect to modifying their drinking habits. Multivariate analyses indicated that a positive history of alcoholism and higher daily consumption were associated with greater readiness to change. CONCLUSIONS: Soon after SCI, most at-risk drinkers are at least considering changes in their alcohol use. This situation may represent an underutilized window of opportunity to implement interventions designed to reduce postinjury alcohol abuse and related impairments. PMID- 9749692 TI - Flexion and traction effect on C5-C6 foraminal space. AB - OBJECTIVE: To determine the effects of cervical flexion and traction on foraminal volume and isthmus area at the C5-C6 foraminal space in cadavers. DESIGN: This study evaluated the foraminal space at C5-C6 in cadaver specimens during flexion and traction of the cervical spine. SETTING: An orthopedic biomechanics laboratory and department of radiology of a university medical center. PATIENTS OR OTHER PARTICIPANTS: Nine cadaver cervical spines, C1 through T3, were used in the study. Superficial tissues were dissected, preserving the ligaments. INTERVENTIONS: Proximal and distal portions of the cadaver spines were potted using bone cement. Spines were mounted and imaged with computed tomography in neutral position, 15 degrees of flexion, and maximum flexion with and without 25lbs of axial traction. MAIN OUTCOME MEASURES: The areas and volumes of the foramen were measured and calculated. RESULTS: Flexion alone significantly increased the foraminal volume and isthmus area at C5-C6. Traction resulted in little additional change. CONCLUSIONS: For cervical spines with mild to moderate degenerative changes at C5-C6, cervical flexion with or without traction produces significant increases in foraminal volume and area at the foraminal isthmus. PMID- 9749694 TI - Short-term attenuation of natural killer cell cytotoxic activity in wheelchair marathoners with paraplegia. AB - OBJECTIVE: To investigate homeostasis of the immune system in athletes with spinal cord injuries during and after racing a wheelchair marathon. DESIGN: The study examined changes in the number and function of natural killer (NK) cells in nine male wheelchair marathon athletes (spinal cord injuries between T5 and T12) who completed the 15th Oita International Wheelchair Marathon Race. Blood samples were obtained the day before, immediately after, and 1 day after the race. Blood samples were also obtained from seven age-matched control subjects with spinal cord injuries but who did not exercise regularly. RESULTS: The number of peripheral leukocytes increased (p < .01) immediately after the race. In contrast, the number of peripheral NK cells and NK cell cytotoxic activity significantly decreased from 310+/-130/microL to 133 +/-61/microL and from 42.6%+/-3.0% to 38.2%+/-3.2%, respectively (mean+/-SD), immediately after the race. Plasma cortisol levels were increased after the race. However, all parameters returned to control levels within 24 hours. Measurements in control subjects did not change throughout the experiment. CONCLUSION: These findings suggest that racing a marathon suppressed peripheral NK cell number as well as NK cell cytotoxic activity in wheelchair athletes and that this was probably mediated by increased postrace cortisol levels. Wheelchair marathon athletes are advised to take extra precaution to avoid infection within 24 hours after racing because of the transient suppression of NK cell cytotoxic activity during this period. PMID- 9749695 TI - Patient satisfaction and rehabilitation services. AB - OBJECTIVE: Despite the widespread use of patient satisfaction measures, there has been only a small amount of research and writing on the topic in rehabilitation. This article reviews selectively the large amount of literature on satisfaction in health care, examines work in rehabilitation settings, and highlights issues in patient satisfaction, given the unique circumstances of rehabilitation services. DATA SOURCES: A Medline search was made of the past 10 years using descriptors related to patient satisfaction, rehabilitation, and selected diagnostic categories. Additional sources came from references on satisfaction accumulated by the author over the past 20 years. STUDY SELECTION: Because of the voluminous literature, findings from existing reviews were emphasized, particularly those using meta-analytic methods. All articles that involved satisfaction in rehabilitation settings were included. DATA SYNTHESIS: Research in health care generally shows high levels of satisfaction. Personal aspects of care, including full communication, are the most important predictors, whereas age, education, and social status show weak relationships with rating levels. Dissatisfied patients tend to seek other providers. Higher satisfaction is associated with patient compliance and better outcomes. Levels of satisfaction are especially high in rehabilitation. CONCLUSIONS: Measures of patient satisfaction with rehabilitation should include items regarding progress and degree of return to independent living. Responses of proxies answering in place of patients should not be regarded as equivalent to patients' opinions. The field is in need of standard, validated measures appropriate for various settings. PMID- 9749696 TI - Long-term cardiac ischemia leading to coronary artery bypass grafting in a tetraplegic patient. AB - With increasing survival in the spinal cord injury (SCI) population, coronary heart disease (CHD) is becoming a leading source of morbidity and mortality. Known risk factors and characteristic signs and symptoms of CHD in the general population may be altered or absent in SCI. This report describes the long-term cardiovascular course and outcome of a man with C6 American Spinal Injury Association Impairment Scale A tetraplegia secondary to a motor vehicle crash. Cardiac risk factors included male gender, mild hypercholesterolemia, and sedentary lifestyle. In retrospect, intermittent tooth pain for 13 years was likely an atypical presentation of angina. Because of severe diffuse coronary and carotid atherosclerotic disease, he underwent simultaneous four-vessel coronary artery bypass graft and carotid endarterectomy. This case demonstrates the challenges to the physiatrist in the diagnosis and management of concurrent CHD and SCI, as well as the benefit of appropriate treatment in individuals with SCI. PMID- 9749697 TI - Bone fracture during electrical stimulation of the quadriceps in a spinal cord injured subject. AB - We report a fracture through the lateral femoral condyle of a paraplegic subject caused by electrical stimulation (ES). The subject was a 50-year-old man who 4 years earlier had sustained a complete spinal cord injury (SCI) at level T6. The fracture occurred during ES-induced measurement of maximal isometric torque of the quadriceps with the knee flexed at an angle of 90 degrees. ES was delivered through surface electrodes with biphasic square wave pulses from a constant current stimulator. The torque was calculated to be 93Nm, corresponding to 20.8kg at the ankle. The regional bone mineral density of the entire lower extremities was .83g/cm2, corresponding to 60% of sex- and age-matched able-bodied reference values. Several factors are suspected to have contributed to the fracture: maximal ES in combination with a muscle spasm, severe osteoporosis, increased muscular strength induced by regular ES cycling (twice a week), and testing position with the knee locked in 90 degrees flexion. The risk of fracture as well as various precautions are discussed and should be taken into consideration in future studies. PMID- 9749698 TI - Low back pain caused by a duodenal ulcer. AB - The common diagnoses in low back pain are lumbar strain, lumbosacral radiculopathy, osteoarthritis, degenerative disc disease, spinal stenosis, and sacroiliac joint dysfunction. Unusual causes of low back pain that have been previously identified include abdominal aortic aneurysms, pelvic neoplasms, and retroperitoneal hemorrhages. This report describes a case of back pain that was apparently caused by a duodenal ulcer. A 54-year-old man with no significant medical history presented with a complaint of mid to low back pain (T10-L2), which was diagnosed as joint dysfunction. A comprehensive treatment program was prescribed and the patient was instructed to return to clinic in 4 weeks. Three weeks later, he experienced a syncopal episode followed by coffee ground emesis. He immediately sought medical attention at an emergency room, where he was admitted to the hospital with a diagnosis of upper gastrointestinal bleed. Esophagogastroduodenoscopy showed a large duodenal ulcer, and the patient underwent vagotomy and pyloroplasty. He returned to his physiatrist's office 3 weeks after hospital discharge with minimal back pain. The cause of the back pain proved to be referred visceral pain from his duodenal ulcer. This case is presented to reemphasize the need to include the uncommon phenomena in the differential diagnosis of low back pain. PMID- 9749699 TI - Respiratory rehabilitation. PMID- 9749700 TI - Needle examination in carpal tunnel syndrome. PMID- 9749701 TI - Lumbar spinal stenosis. PMID- 9749702 TI - The Eph receptor tyrosine kinases and the ephrins--roles in nervous system development. PMID- 9749703 TI - The intermediate stage and paradoxical sleep in the rat: influence of three generations of hypnotics. AB - Paradoxical sleep in the rat, cat and mouse is preceded and sometimes followed by a short-lasting intermediate stage characterized by high-amplitude anterior cortex spindles and low-frequency hippocampal theta rhythm. Several neurophysiological arguments suggest that the intermediate stage corresponds to a brief functional disconnection of the forebrain from the brainstem. This paper is devoted to the review of quantitative and qualitative influences of three generations of hypnotics on the intermediate stage-paradoxical sleep couple. Barbiturates, first-generation hypnotics, extend the intermediate stage at the expense of paradoxical sleep. Three benzodiazepines are compared, two with a short half-life (triazolam and midazolam) and one with a long half-life (diazepam). They also decrease sleep occurrence latency and increase the intermediate stage at the expense of paradoxical sleep, except for midazolam, which increases both the intermediate stage and paradoxical sleep at low dose. Zolpidem and zopiclone, hypnotics of third generation, decrease paradoxical sleep but the intermediate stage never substitutes for paradoxical sleep. The results are discussed in relationship to the functional aspects of this turning-point period of sleep. PMID- 9749704 TI - Exocytosis: SNAREs drum up! PMID- 9749705 TI - Enhanced neurotransmitter release is associated with reduction of neuronal branching in a Drosophila mutant overexpressing frequenin. AB - Frequenin is a Drosophila Ca2+ binding protein whose overexpression causes a chronic facilitation of transmitter release at the larval neuromuscular junction and multiple firing of action potentials. These functional abnormalities are similar to those found in other hyperexcitable mutants (Shaker, ether-a-gogo, Hyperkinetic) which, in turn, exhibit increased branching at the motor nerve endings. We report here that mutants which overexpress frequenin have motor nerve terminals with reduced number and length of branches as well as number of synaptic boutons. Similar defects are observed in transgenic flies which have additional copies of the frequenin gene indicating that the phenotype can be adscribed to the overexpression of the protein. The ultrastructure of boutons, however, appears indistinguishable from wild type. In addition, we show here that frequenin overexpression leads also to a down regulation of Shaker proteins expression. The contrast between the observations in frequenin and the other hyperexcitable mutants indicates that nerve terminal morphology and enhanced transmitter release do not have a direct causal relationship. PMID- 9749706 TI - Modulation of bradykinin-induced mechanical hyperalgesia in the rat by activity in abdominal vagal afferents. AB - Bradykinin-induced plasma extravasation and mechanical hyperalgesia are sympathetic-dependent components of inflammation. Noxious stimulation has been found to inhibit bradykinin-induced plasma extravasation by activating the hypothalamo-pituitary-adrenal axis. The sensitivity of this nociceptive neuroendocrine feedback control of inflammation is modulated by activity in subdiaphragmatic vagal afferents. In the present study, we tested the hypothesis that activity in the subdiaphragmatic vagus also modifies bradykinin-induced mechanical hyperalgesia in the rat, using the Randall-Selitto method. Following subdiaphragmatic vagotomy, the baseline paw-withdrawal threshold to mechanical stimulation decreased and bradykinin-induced mechanical hyperalgesia was enhanced. Mechanical hyperalgesia produced by prostaglandin E2, a direct-acting hyperalgesic agent, was not significantly affected by vagotomy. The effect of subdiaphragmatic vagotomy on bradykinin-induced hyperalgesia, but not on baseline paw-withdrawal threshold, was mimicked by coeliac branch vagotomy. Indomethacin blocked the hyperalgesia in normal rats, but not in vagotomized rats, suggesting that bradykinin-induced hyperalgesia in normal rats is mediated by prostaglandins, whose role was unexpectedly diminished after vagotomy. Bradykinin induced hyperalgesia in normal rats was abolished by lumbar sympathectomy but not by sympathetic decentralization (cutting the preganglionic axons). In rats that were both vagotomized and sympathectomized, hyperalgesia induced by low-dose bradykinin was no longer present. These results demonstrate that vagotomy induces a decrease in baseline mechanical paw-withdrawal threshold and an enhancement of bradykinin-induced mechanical hyperalgesia and suggest that these phenomena are generated by actions in peripheral tissues. PMID- 9749707 TI - Neuronal expression of inducible nitric oxide synthase after oxygen and glucose deprivation in rat forebrain slices. AB - Nitric oxide (NO) overproduction has been postulated to contribute significantly to ischaemia-reperfusion neurotoxicity. Inducible or type II NO synthase (iNOS) synthesizes NO in large quantities for long periods of time. Therefore we investigated the expression and localization of iNOS after oxygen and glucose deprivation in rat forebrain slices. In this experimental model, calcium independent NOS activity reached a maximum 180 min after the end of a 20 min oxygen-glucose deprivation period. During the same period of time, the calcium independent activity was absent in control forebrain slices. To test whether this calcium-independent NOS activity was due to the expression of iNOS, the effects of the addition of dexamethasone, cycloheximide and pyrrolidine dithiocarbamate were determined. All of them inhibited the induction of the calcium-independent NOS activity measured in the rat forebrain slices after oxygen and glucose deprivation. Furthermore, oxygen and glucose deprivation caused the expression of the gene encoding iNOS in rat forebrain slices, as assessed by the detection of iNOS message and protein in these samples. A sixfold increase in the iNOS mRNA levels was observed at 180 min and the time-course of the expression of iNOS mRNA was in agreement with the temporal profile of iNOS enzymatic activity. Immunohistochemistry analysis revealed that iNOS was highly expressed in neurones, astrocytes and microglial cells. These results demonstrate for the first time that iNOS is expressed in neurones after oxygen and glucose deprivation, and that this expression occurs in short periods of time. These findings suggest that NO can play an important pathogenic role in the tissue damage that occurs after cerebral ischaemia. PMID- 9749708 TI - Footshock stress but not contextual fear conditioning induces long-term enhancement of auditory-evoked potentials in the basolateral amygdala of the freely behaving rat. AB - In this study, rats were bilaterally implanted with electrodes in the amygdala for chronic recording. Auditory click stimulation evoked in the basolateral nucleus a field potential characterized by three positive components: P1, P2 and P3 (peak latencies around: 10, 20 and 30 ms, respectively) which were each followed by three negative components: N1, N2 and N3 (peak latencies around: 13, 30 and 50 ms, respectively). Animals were divided into three groups (context same, context-different and control). Following footshock administration, animals were either re-exposed to the same conditioning chamber (context-same group) or placed in a different context (context-different group) for electrophysiological and behavioural (evaluation of freezing response) recordings. The two early positive-negative complexes (P1-N1 and P2-N2) increased in amplitude from 2 min to 24 h following footshock in both context-same and context-different groups. No significant difference was observed between these two groups. The demonstration of significantly larger freezing responses in context-same subjects on exposure to the aversive conditioned environment indicated that this similarity of effects was not due to lack of conditioning of context under the experimental conditions chosen. We conclude that footshock stress produces general long-lasting changes in amygdala auditory field potentials that are not significantly affected by contextual fear conditioning. PMID- 9749709 TI - The rhythmicity of cells of the medial septum/diagonal band of Broca in the awake freely moving rat: relationships with behaviour and hippocampal theta. AB - Chronic extracellular recordings were obtained from cells of the medial septum and diagonal band of Broca in rats performing a simple behavioural task. The cells were found to display a variety of bursting patterns phase-locked to hippocampal theta rhythm to a greater or lesser degree. Among phase-locked cells, no systematic distribution in preferential phase could be found, and these cells were shown to maintain their preferential phase for extended periods. Cells were classified into those which showed signs of a broadening of the repolarization phase of their action potential ('inflected': putative cholinergic) and those without ('non-inflected': putative GABAergic). Non-inflected cells tended to fire rhythmic bursts while inflected cells mostly fired in an irregular fashion, although still significantly phase-locked to hippocampal theta. In neither population did the phase-locked cells show any coherent distribution of their preferential phase. Sixty-five per cent of the rhythmically bursting cells showed a significant correlation between the interburst frequency and the animal's running speed. Five cells displayed rhythmic activity only when the rat ran in a specific direction. These results have implications for models of septohippocampal function and the effects of variable septal rhythmicity on the production of hippocampal theta rhythm. PMID- 9749710 TI - Selective scarcity of NMDA receptor channel subunits in the stratum lucidum (mossy fibre-recipient layer) of the mouse hippocampal CA3 subfield. AB - Hippocampal synapses express two distinct forms of the long-term potentiation (LTP), i.e. NMDA receptor-dependent and -independent LTPs. To understand its molecular-anatomical basis, we produced affinity-purified antibodies against the GluRepsilon1 (NR2A), GluRepsilon2 (NR2B), and GluRzeta1 (NR1) subunits of the N methyl-D-aspartate (NMDA) receptor channel, and determined their distributions in the mouse hippocampus. Using NMDA receptor subunit-deficient mice as the specificity controls, section pretreatment with proteases (pepsin and proteinase K) was found to be very effective to detect authentic NMDA receptor subunits. As the result of modified immunohistochemistry, all three subunits were detected at the highest level in the strata oriens and radiatum of the CA1 subfield, and high levels were also seen in most other neuropil layers of the CA1 and CA3 subfields and of the dentate gyrus. However, the stratum lucidum, a mossy fibre-recipient layer of the CA3 subfield, contained low levels of the GluRepsilon1 and GluRzeta1 subunits and almost excluded the GluRepsilon2 subunit. Double immunofluorescence with the AMPA receptor GluRalpha1 (GluR1 or GluR-A) subunit further demonstrated that the GluRepsilon1 subunit was colocalized in a subset, not all, of GluRalpha1 immunopositive structures in the stratum lucidum. Therefore, the selective scarcity of these NMDA receptor subunits in the stratum lucidum suggests that a different synaptic targeting mechanism exerts within a single CA3 pyramidal neurone in vivo, which would explain contrasting significance of the NMDA receptor channel in LTP induction mechanisms between the mossy fibre-CA3 synapse and other hippocampal synapses. PMID- 9749711 TI - Modification of parallel activity elicited by propagating bursts in developing networks of rat cortical neurones. AB - Networks of cultured cortical neurones exhibit regular, synchronized, propagating bursts which are synaptically mediated, and which are hypothesized to play a part in activity-dependent formation of connections during development in vivo. The relationship between the strength of synaptic connections and the characteristics of synchronized propagating bursting, however, is unclear. Modification of synchronized activity in cortical cultures in response to electrical stimulation was examined using multisite electrode array recording. By measuring the response of the network to weak, localized, test stimulation (TS), we observed a potentiation of activity following a relatively stronger inducing stimulation (IS). This potentiation was evident as an increased probability of eliciting bursts by TS, an increased frequency of spontaneous bursts and number of spikes per burst, and increased speed of burst propagation, and it lasted for at least 20 min. Changing the parameters of IS revealed that high frequency tetanic stimulation is not necessary to induce potentiation, while it is essential for IS to produce a regeneratively propagating burst. The results provide a direct demonstration of modification of both the spatial and temporal characteristics of synchronized network activity, and suggest an important physiological role for propagating synchronized bursting, as a mechanism for inducing plastic modifications in the developing cortex. PMID- 9749712 TI - Long-lasting enhanced expression in the rat hippocampus of NMDAR1 splice variants in a kainate model of epilepsy. AB - Chronic epilepsy is associated with increased excitability which may result from abnormal glutamatergic synaptic transmission involving altered properties of N methyl-D-aspartate (NMDA) receptors. To date two gene families encoding NMDA receptor subunits have been cloned, NR1 and NR2. Eight NR1 mRNAs are generated by alternative splicing of exons 5, 21 and 22; the NR1-1 to NR1-4 C-terminal variants exist in the a or b version depending on the presence or absence of the domain encoded by exon 5. Epilepsy was induced in rats by unilateral intra amygdalar injection of kainate and animals were killed from 6 h to 4 months following the injection. Increased NR1 mRNA levels were observed during status epilepticus (6-24 h after the injection), both psilateral and contralateral, while a second wave of NMDAR1 mRNA increase occurred in chronic epileptic animals, between 21 days and 4 months following kainate injection. Our data show: (i) a permanent increase of the NR1-2a and NR1-2b mRNA species (containing exon 22) in all hippocampal fields, both ipsilateral and contralateral, and (ii) an increase of the NR1-3 (a and b) mRNAs (containing exon 21) in the ipsilateral CA1, and NR1-3a mRNA in the ipsilateral dentate gyrus. No long-term changes were observed for the NR1-1 and NR14 splice variants. In the ipsilateral CA3 area a globally decreased mRNA expression was associated with neuronal loss. A possible contribution to the maintenance of the epileptic state by an increased expression of NMDA receptors is discussed. PMID- 9749713 TI - CRE and TRE sequences of the rat tyrosine hydroxylase promoter are required for TH basal expression in adult mice but not in the embryo. AB - Tyrosine hydroxylase (TH), the rate-limiting enzyme in the biosynthesis of catecholamine neurotransmitters, is expressed in a restricted number of areas, and subject to numerous regulations during development and in adulthood. Two transcription factor binding sites present in the proximal region of the TH gene, the TPA-responsive element (TRE) and the c-AMP responsive element (CRE), have been shown to play important roles in TH gene regulation in vitro. In order to elucidate in vivo the role of these two sites, we produced transgenic mice bearing a 5.3-kb fragment from the 5' flanking sequence of the TH gene with mutations in either the CRE-or TRE-sites. Using the intact 5.3-kb fragment fused to two different reporter genes (HSV1-tk and lacZ), we show that this promoter fragment is able to specifically direct expression in catecholaminergic tissues both in adult mice and embryos. Interestingly, the CRE- and TRE-mutated transgenes were not expressed in adult mice, contrary to the situation in embryos where they were specifically expressed in catecholaminergic regions. These results demonstrate that the CRE and TRE play an essential role in basal TH expression in adult tissues in vivo. Moreover, they suggest that distinct transcription factors are involved in TH regulation in developing and adult tissues. In support of this, gel mobility shift experiments revealed a complex present only in embryonic tissues. Taken together, these data highlight the diversity of the mechanisms underlying the establishment and maintenance of the catecholaminergic phenotype. PMID- 9749714 TI - High-affinity copper block of GABA(A) receptor-mediated currents in acutely isolated cerebellar Purkinje cells of the rat. AB - The actions of Cu2+ ions on GABAA receptor-mediated currents in acutely isolated Purkinje cells from rat cerebellum were studied using the whole-cell patch-clamp technique and a rapid perfusion system. Bath application of Cu2+ reduced currents induced by 2 microM gamma-aminobutyric acid (GABA) in a concentration-dependent manner with an IC50 of 35 nM. The Cu2+-induced block of GABA responses was not voltage-dependent. Increasing the GABA concentration (from 2 to 50 microM) decreased the blocking effect of Cu2+. Dose-response analysis for activation of GABAA receptors revealed a twofold decrease in apparent affinity for GABA in the presence of 0.1 microM Cu2+. Recovery from the block required several minutes after removal of Cu2+ from the medium. The block was removed by histidine, which preferentially forms complexes with Cu2+, or by other chelating substances. Application of 10 microM histidine immediately before application of 2 microM GABA completely relieved the block of GABA responses produced by 0.1 microM Cu2+. The effect of histidine was concentration-dependent with an EC50 of 0.75 microM. The results demonstrate that Cu2+ is a potent inhibitor of GABA-evoked responses in rat Purkinje cells. Copper may be an endogenous synaptic modulating factor. Cu2+ toxicity, notably in Wilson's disease, could result to some extent from chronic GABAA receptor blockade. PMID- 9749715 TI - Development and characterization of antibodies directed against the mouse D4 dopamine receptor. AB - Polyclonal antibodies against the mouse D4 dopamine receptor have been developed in order to investigate the anatomical localization of this receptor in the mouse brain. Two antibodies were generated against specific peptides corresponding to predicted extracellular and intracellular regions of the D4 protein. Specificity of these antibodies was demonstrated on human embryonic kidney 293 (HEK 293) cells transfected with different dopamine receptor subtypes; immunoreactivity was detected only in cells transfected with the mouse D4 dopamine receptor cDNA. Following in vitro transcription/translation of the mouse D4 cDNA, a single protein band of 36 kDa was selectively immunoprecipitated with the anti-D4 antibodies. The antibodies also detected a single protein of 36 kDa in Western blot of HEK 293 cells transiently transfected with the mouse D4 receptor. These antibodies were able to detect the D4 receptor in several regions of the mouse brain. In the regions examined, D4 immunoreactivity was found in neurones located in layers II-VI of the frontal and piriform cortices, with the highest concentration in layer II; in scattered neurones in the caudate putamen and in larger neurones in the globus pallidus. In all experiments, both antibodies exhibit the same specificity, and all immunoreactivity could be abolished by preincubation with the corresponding peptide antigen. PMID- 9749716 TI - Reversible impairment of long-term potentiation in transgenic Cu/Zn-SOD mice. AB - Copper/zinc superoxide dismutase (CuZn-SOD) is a key enzyme in the metabolism of oxygen free radicals. The gene encoding CuZn-SOD resides on human chromosome 21 and is overexpressed in Down syndrome (DS) patients. Overexpression of CuZn-SOD in transgenic (Tg) mice and cultured cells creates chronic oxidative stress leading to enhanced susceptibility to degeneration and apoptotic cell death. We have now found that three lines of Tg-CuZn-SOD mice, one of which also overexpresses S100beta, a glial calcium binding protein, are deficient in spatial memory. Furthermore, hippocampal slices taken from these mice have an apparently normal synaptic physiology, but are impaired in the ability to express long-term potentiation (LTP). This effect on hippocampal LTP was abrogated by treatment of slices with the H2O2 scavenger catalase or the antioxidant N-t-butyl phenylnitrone (BPN). It is proposed that elevated CuZnSOD causes an increase in tetanic stimulation-evoked formation of H2O2 which leads to diminished LTP and cognitive deficits in these mice. PMID- 9749717 TI - Developmental regulation of T-, N- and L-type calcium currents in mouse embryonic sensory neurones. AB - We investigated the development of a low (T-type) and two high voltage-activated (N- and L-type) calcium channel currents in large diameter dorsal root ganglion neurones acutely isolated from embryonic mice using the whole-cell patch-clamp technique. The low and high voltage-activated barium currents (LVA and HVA) were identified by their distinct threshold of activation and their sensitivity to pharmacological agents, dihydropyridines and omega-conotoxin-GVIA, at embryonic day 13 (E13), E15 and E17-18, respectively, before, during and after synaptogenesis. The amplitude and density of LVA currents, measured during a -40 mV pulse from a holding potential of -100 mV, increased significantly between E13 and E15, and remained constant between E15 and E17-18. The density of global HVA current, elicited by 0 mV pulse, increased between E13 and E15/E17-18. The density of the N-type current studied by the application of omega-conotoxin-GVIA (1 microM) increased significantly between E13 and E15/E17-18. The use of the dihydropyridine nitrendipine (1 microM) revealed that the density of L-type current remained constant at each stage of development. Nevertheless, application of dihydropyridine Bay K 8644 (3 microM) demonstrated a significant slowing of the deactivation tail current between embryonic days 13 and 15, which may reflect a qualitative maturation of this class of calcium channel current. The temporal relationship between the changes in calcium channel pattern and the period of target innervation suggests possible roles of T-, N- and L-type currents during developmental key events such as natural neurone death and onset of synapse formation. PMID- 9749718 TI - Evidence for the involvement of corticotrophin-releasing hormone in the pathogenesis of traumatic brain injury. AB - The aim of this study was to investigate the role of the neuropeptide corticotrophin-releasing hormone (CRH) in neurodegeneration induced by traumatic brain injury, using a well characterized model of lateral fluid percussion injury in male, Sprague-Dawley rats. In the first series of experiments, CRH gene expression was assessed by in situ hybridization after traumatic brain injury. A bilateral increase in CRH mRNA in the paraventricular nucleus was observed in rats subjected to traumatic brain injury compared with sham-operated controls. A maximal (40%) increase in hybridization signal was detected 2 h after trauma compared with control rat brains. In addition, marked induction of CRH transcripts was found in the ipsilateral amygdala after trauma, but no increase was detected in the ipsilateral cortex around the area of damage. In a separate experiment, the effects of the CRH antagonist, D-Phe CRH(12-41) (25 microg total dose), or appropriate vehicle injected intracerebroventricularly, was tested on infarct volume caused by brain injury. Repeated intracerebroventricular injection of D-Phe CRH(12-41) significantly reduced, by 45%, the volume of cortical damage in injured rats compared with vehicle-treated, trauma animals. The rapid upregulation of CRH gene expression in the paraventricular nucleus and amygdala following lateral fluid percussion injury and the marked neuroprotection achieved by inhibiting CRH action suggest that CRH is involved directly in the pathogenesis of traumatic brain injury. This observation may have important implications for the development of novel therapeutic strategies for treating the neurological consequences of brain trauma and related conditions. PMID- 9749719 TI - Influence of acetylcholinesterase on embryonic spinal rat motoneurones growth in culture: a quantitative morphometric study. AB - Rat spinal motoneurones sampled at day embryonic 15 were purified using a Nycodenz gradient and cultured in defined medium, during 7 days, on glass coverslips coated with poly-L-lysine and laminine. Purified acetylcholinesterase (AChE), ecothiopate, BW 284C51 and fasciculin II, inhibitors of either the catalytic or peripheral site of AChE, were added to the defined medium. Morphological changes of spinal motoneurones were measured using a statistical quantitative morphometric method, allowing the determination of various parameters such as the number of neurites and bifurcations, the length of neurites, the surface and spreading index. Presence of AChE in the medium (4 units/mL) increases the surface and the total length of neurites and axons without any change in the spreading index. When spinal motoneurones were cultured on AChE coated substrate, neurones rapidly migrate and form clusters. Addition of ecothiopate (10(-6) M) in the medium, which selectively blocks the catalytic site of AChE, leads to a slight increase in the number of primary neurite and a decrease of the spreading index during the three first days in culture. BW 284C51 (10(-5) M) which blocks the catalytic site but also affect the peripheral one, significantly reduces the number of primary neurites and increases the number of bifurcations. Fasciculin II, a potent blocker (10(-9)M) of the AChE peripheral site induces a decrease of both primary neurites and bifurcations with a significant increase of the length and growth velocity of the axon, giving a drastic enhancement of the spreading index. These phenomena are discussed in terms of catalytic and non-catalytic function of AChE, including the involvement of the enzyme in adhesive processes, occurring during growth and differentiation of spinal motoneurones. PMID- 9749720 TI - Peptidergic innervation and the nicotinic acetylcholine receptor in the primate basal nucleus. AB - Peptidergic innervation and localization of the neuronal nicotinic acetylcholine receptor (nAChR) was studied in the basal forebrain of Macaca fascicularis in order to provide microstructural proofs for the theory (Changeux et al., 1992) that calcitonin gene-related peptide (CGRP) is responsible for the maintenance of the acetylcholine receptor. Distribution and localization of five neuropeptides, namely substance P (SP), CGRP, neuropeptide Y (NPY), vasoactive intestinal polypeptide (VIP) neurotensin (NT), and the neuropeptides parvalbumin (PV) and the alpha-bungarotoxin- (alpha-BTX-) binding protein was studied by means of light- and electron microscopic pre-embedding immunocytochemistry. Immunohistochemical double staining revealed that large cholinergic principal nerve cells in the basal forebrain, corresponding to cell group Ch4 constituting Meynert's basal nucleus (BNM), and exerting intense choline acetyltransferase (ChAT) immunoreactivity, are synaptically innervated by axons displaying CGRP immunoreactivity. While SP, NPY, PV and CGRP establish dense networks in BNM, innervation by NT and VIP is sparse. Biotinylated alpha-BTX visualizes beaded axons that surround dendrites and perikarya of cholinergic principal cells. Electron microscopic organization of the neuropil in BNM is characterized by a glomerular (or rather cartridge-like) arrangement of axons surrounding dendrites of non-cholinergic principal nerve cells. At least one of the axons establishing the glomerulus (cartridge) exerts CGRP immunopositivity while alpha-BTX immunopositive axons, presynaptic to dendrites of principal cells, are attached to the glomeruli (cartridges) from outside. As alpha-BTX-binding indicates localization of the alpha7 subunit of the neuronal nAChR, the microtopographical arrangement supports the idea that, in a manner similar to that in the neuromuscular junction, CGRP might contribute to the maintenance of nAChR also in BNM. Our results suggest that presynaptic nAChR-s are involved in the regulation of acetylcholine release from a feed-forward amplification mechanism of cholinergic principal cells of BNM. PMID- 9749721 TI - Distribution of L-type calcium channels in rat thalamic neurones. AB - One major pathway for calcium entry into neurones is through voltage-activated calcium channels. The distribution of calcium channels over the membrane surface is important for their contribution to neuronal function. Electrophysiological recordings from thalamic cells in situ and after acute isolation demonstrated the presence of high-voltage activated calcium currents. The use of specific L-type calcium channel agonists and antagonists of the dihydropyridine type revealed an about 40% contribution of L-type channels to the total high-voltage-activated calcium current. In order to localize L-type calcium channels in thalamic neurones, fluorescent dihydropyridines were used. They were combined with the fluorescent dye RH414, which allowed the use of a ratio technique and thereby the determination of channel density. The distribution of L-type channels was analysed in the three main thalamic cell types: thalamocortical relay cells, local interneurones and reticular thalamic neurones. While channel density was highest in the soma and decreased significantly in the dendritic region, channels appeared to be clustered differentially in the three types of cells. In thalamocortical cells, L-type channels were clustered in high density around the base of dendrites, while they were more evenly distributed on the soma of interneurones. Reticular thalamic neurones exhibited high density of L-type channels in more central somatic regions. The differential localization of L-type calcium channels found in this study implies their predominate involvement in the regulation of somatic and proximal dendritic calcium-dependent processes, which may be of importance for specific thalamic functions, such as those mediating the transition from rhythmic burst activity during sleep to single spike activity during wakefulness or regulating the relay of visual information. PMID- 9749722 TI - A role for BDNF in early postnatal rat vestibular epithelia maturation: implication of supporting cells. AB - The early development of the inner ear is largely determined by two members of the neurotrophic family: brain-derived neurotrophic factor (BDNF) and neurotrophin 3 (NT-3). Little information is available on the role of these neurotrophins during the late stages of vestibular development in the rat which take place during the first postnatal weeks. At this period where terminal synaptogenesis and maturation occur, we have investigated the expression and the activity of BDNF, the most important neurotrophin in the vestibular system. Using different experimental approaches, we show that BDNF is released by vestibular epithelia on postnatal day 3 (P3) and continues to have a trophic effect on vestibular neurones in vitro. Immunocytochemistry coupled to confocal microscopy revealed a remarkable evolution in BDNF localization during later stages of development. Whereas BDNF is present in both supporting cells and hair cells at P3, its distribution gradually changed and is highly compartmentalized within the upper part of supporting cells at P8 and P15. In parallel, we observed the presence of a truncated form of the BDNF receptor in sensory hair cells. These results suggest an original role for supporting cells, which could be involved in the release of BDNF during the late stages of synaptogenesis in mammalian vestibular epithelia. In particular, BDNF could participate to the set up of the calyx, a specific nerve structure surrounding type I vestibular hair cells. PMID- 9749723 TI - Schwann cells genetically modified to secrete human BDNF promote enhanced axonal regrowth across transected adult rat spinal cord. AB - The infusion of BDNF and NT-3 into Schwann cell (SC) grafts promotes regeneration of brainstem neurones into the grafts placed in adult rat spinal cord transected at T8 (Xu et al., 1995b). Here, we compared normal SCs with SCs genetically modified to secrete human BDNF, grafted as trails 5 mm long in the cord distal to a transection site and also deposited in the transection site, for their ability to stimulate supraspinal axonal regeneration beyond the injury. SCs were infected with the replication-deficient retroviral vector pL(hBDNF)RNL encoding the human preproBDNF cDNA. The amounts of BDNF secreted (as detected by ELISA) were 23 and 5 ng/24 h per 106 cells for infected and normal SCs, respectively. Biological activity of the secreted BDNF was confirmed by retinal ganglion cell bioassay. The adult rat spinal cord was transected at T8. The use of Hoechst prelabelled SCs demonstrated that trails were maintained for a month. In controls, no SCs were grafted. One month after grafting, axons were present in SC trails. More 5 HT-positive and some DbetaH-positive fibres were observed in the infected vs. normal SC trails. When Fast Blue was injected 5 mm below the transection site (at the end of the trail), as many as 135 retrogradely labelled neurones could be found in the brainstem, mostly in the reticular and raphe nuclei (normal SCs, up to 22, mostly in vestibular nuclei). Numerous neurones were labelled in the ventral hypothalamus (normal SCs, 0). Also, following Fast Blue injection, a mean of 138 labelled cells was present in dorsal root ganglia (normal SCs, 46) and spinal cord (39 vs. 32) rostral to the transection. No labelled spinal neurones rostral to the transection were seen when SCs were not transplanted. Thus, the transplantation of SCs secreting increased amounts of BDNF improved the regenerative response across a transection site in the thoracic cord. Moreover, the enhanced regeneration observed with infected SCs may be specific as the largest response was from neurones known to express trkB. PMID- 9749724 TI - Comparing the effects of selective cingulate cortex lesions and cingulum bundle lesions on water maze performance by rats. AB - The ability of rats to learn the location of a hidden platform in a swim maze was compared in animals with excitotoxic lesions of the anterior or posterior (retrosplenial) cingulate cortex or radiofrequency lesions of the cingulum bundle or fimbria-fornix. Performance of this allocentric spatial task was unaffected by the posterior cingulate cortex lesions, while anterior cingulate cortex damage produced only a mild acquisition deficit. Transection of the fornix and lesions of the cingulum bundle produced similar patterns of impairment on initial acquisition, but the cingulum bundle lesions had less effect on reversal of the task. The results from the water maze, and from a subsequent T-maze alternation task, indicate that cingulum bundle lesions can produce a spatial deficit that is similar, but milder, to that observed after fornix transection. The results of the excitotoxic lesions suggest that previous studies examining conventional cingulate lesions may have been influenced by damage to adjacent fibre tracts, such as the cingulum bundle. PMID- 9749725 TI - N-syndecan and HB-GAM (heparin-binding growth-associated molecule) associate with early axonal tracts in the rat brain. AB - Heparin-Binding Growth-Associated Molecule (HB-GAM)/pleiotrophin is an 18 kDa extracellular matrix- and cell-surface-associated protein shown to enhance neurite outgrowth of perinatal forebrain neurones in vitro. The heparan sulphate proteoglycan N-syndecan (Raulo et al., 1994) has been isolated as a receptor/coreceptor for the HB-GAM. We have investigated, whether HB-GAM and N syndecan could have a similar role in neurite outgrowth and axon guidance in early axonal tracts of brain. In the present study N-syndecan was found to be spatiotemporally associated with the developing axonal tracts already on embryonic day 9 in rat, as revealed by coexpression with class III beta-tubulin, which is one of the earliest neuronal markers (Easter et al., 1993; Brittis et al., 1995). Later, N-syndecan and HB-GAM were detected in the first afferent serotonergic projections arising from the pontine raphe nuclei. The expression pattern of HB-GAM peaked in the developing rhombencephalon at embryonic stage (E) 13-14. At the same time, N-syndecan was expressed in the developing raphe neurones growing neurites towards the diencephalon along HB-GAM immunoreactive pathways. When rhombencephalic neurones were cultured on decreasing concentrations of substrate-bound HB-GAM, E13 neurones showed a significantly better neurite outgrowth response than E11, E16 or E18 neurones. The neurite outgrowth of raphe neurones in vitro was inhibited by adding soluble heparin or N syndecan into the culture medium, whereas addition of chondroitin sulphate had no effect. In a simple pathway assay, E13 raphe neurones selectively preferred attaching and growing neurites on pathways containing HB-GAM as compared with regions containing either laminin or fibronectin alone. Our results suggest that HB-GAM may function as a developmentally regulated cue for rhombencephalic neurones that possess N-syndecan on their cell membrane. PMID- 9749726 TI - The cerebral metabolic effects of manipulating glutamatergic systems within the basal forebrain in conscious rats. AB - N-methyl-D-aspartate (NMDA) and non-NMDA receptor-mediated manipulations of the cortical cholinergic input arising from the basal forebrain differentially affect cognitive function. We used [14C]-2-deoxyglucose autoradiography in conscious rats to map the effects of excitatory amino acid agonist infusions into the nucleus basalis magnocellularis (NBM) on cerebral functional activity, as reflected by local rates of glucose utilization. Acute stimulation of NBM neurones by local infusion of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA), 15 min before glucose use measurement, resulted in glucose use reductions in nine cortical regions innervated by NBM efferents including prefrontal, frontal, sensorimotor and cingulate cortices. NMDA infusions altered glucose use in two cortical areas. Both AMPA and NMDA markedly increased glucose use in the striatum and globus pallidus, with concomitant perturbations in striato-pallidal projection targets including the substantia nigra, entopeduncular nucleus, subthalamic nucleus and lateral habenular nucleus. In contrast, the GABAA agonist muscimol did not affect glucose use in the NBM or neocortical regions, but induced glucose use increases in several subcortical nuclei including the substantia nigra and entopeduncular nucleus. The delayed effects of excitotoxic lesions were assessed 3 weeks after basal forebrain infusions of AMPA, NMDA, ibotenate or quisqualate. Statistically significant glucose use changes only occurred in the hypothalamus after NMDA, and the NBM after ibotenate infusions, although reduced cortical metabolism was apparent following AMPA-induced lesions of the NBM. Results support a dissociation between the functional sequelae of NMDA and non-NMDA receptor-mediated events in the basal forebrain, and long-term compensatory functional adaptation following cortical denervation. PMID- 9749727 TI - Kinetics of AMPA-type glutamate receptor channels in rat caudate-putamen neurones show a wide range of desensitization but distinct recovery characteristics. AB - alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-type glutamate receptor channels of rat caudate-putamen neurones were studied by ultrafast application of agonists to outside-out vesicles taken from medium-sized spiny neurones in thin slices. Upon application of 10 mM glutamate for 50 ms, fast rising and desensitizing currents were activated. Ten to 90% rise time values were approximately 0.5 ms. Dose-response studies revealed an EC50 of 0.63 mM glutamate. In double logarithmic coordinates, the curve had a maximal slope between 1.33 and 1.85 at low concentrations, indicating at least two binding sites for glutamate. Rise time increased with low agonist concentrations, whereas desensitization kinetics showed only a weak dependence on concentration. The time constant of desensitization was fitted with one exponential and ranged between 2 and 11 ms, with a mean of 6.19+/-2.31 ms (n = 239). Following brief glutamate pulses (1 ms) currents decayed with time constants of 2.7+/-0.23 ms (n = 12). Recovery from desensitization was investigated by double-pulse experiments. Recovery time constants fell in two subgroups with respective mean values of 110.6+/-14.2 ms (n = 8) and 288.6+/-33.2 ms (n = 8). By adding low glutamate concentrations to the bath solution, predesensitization of AMPA-type receptors without channel opening could be shown. A 50% reduction in control amplitude was achieved with 5.2+/-2.1 microM (n = 22) glutamate in the background. We hypothesize a circular reaction scheme with at least two binding sites for glutamate to describe activation, desensitization and recovery from desensitization in rat caudate-putamen neurones. PMID- 9749729 TI - Focal ischaemia of the rat brain elicits an unusual inflammatory response: early appearance of CD8+ macrophages/microglia. AB - Cerebral ischaemia leads to profound glial activation and leukocyte infiltration into the infarct area. In this study, we provide evidence for a dual macrophage response in focal ischaemic lesions of the rat brain. We show that a considerable proportion of macrophages in the ischaemic lesions express the CD8alphabeta heterodimer to date only described on CD8+ T cells. As known from other lesion paradigms, CD4+ macrophages were also present. Interestingly, CD8- and CD4 expressing macrophages formed two non-overlapping subpopulations. CD8+ macrophages reached their maximum during the first week with pronounced downregulation thereafter whereas CD4+ cells persisted at high levels into the second week. In contrast to cerebral ischaemia, macrophages in the spleen and in Wallerian degeneration after optic nerve axotomy expressed CD4, but not CD8. In experimental autoimmune encephalomyelitis, CD8 was mainly associated with T cells and very weakly detectable on some ramified cells resembling activated microglia. In conclusion, we show that cerebral ischaemia triggers an unusual inflammatory response characterized by the appearance of CD8+/CD4- macrophages that might exert specific functions in the pathogenesis of ischaemic brain damage. PMID- 9749728 TI - Cytokines promote the survival of mouse cranial sensory neurones at different developmental stages. AB - To investigate when the neurotrophic cytokines ciliary neurotrophic factor (CNTF), leukaemia inhibitory factor (LIF), oncostatin-M (OSM), interleukin-6 (IL 6) and cardiotrophin-1 (CT-1) act on developing sensory neurones and whether they co-operate with neurotrophins in regulating neuronal survival, we studied the in vitro trophic effects of these factors on two well-characterized populations of cranial sensory neurones at closely staged intervals throughout embryonic development. The cutaneous sensory neurones of the trigeminal ganglion, which show an early, transient survival response to BDNF and NT3 before becoming NGF dependent, were supported by CNTF, LIF, OSM and CT-1 during the late fetal period, several days after the neurones become NGF-dependent. At this stage of development, these cytokines promoted the survival of a subset of NGF-responsive neurones. The enteroceptive neurones of the nodose ganglion, which retain dependence on BDNF throughout fetal development, were supported throughout their development by CNTF, LIF, OSM and CT-1, and displayed an additional survival response to IL-6 in the late fetal period. These findings indicate that populations of sensory neurones display different developmental patterns of cytokine responsiveness and show that embryonic trigeminal neurones pass through several phases of differing neurotrophic factor survival requirements. PMID- 9749730 TI - Behavioural effects of subthalamic nucleus lesions in the hemiparkinsonian marmoset (Callithrix jacchus). AB - Recent studies in non-human primates support a role for the subthalamic nucleus in the expression of parkinsonian symptomatology, and it has been proposed that subthalamic lesions may provide a surgical treatment for the symptoms of Parkinson's disease in humans. We have applied a broad range of behavioural tests to characterize the effects of lesions of the subthalamic nucleus on parkinsonian symptoms in the unilateral 6-hydroxydopamine (6-OHDA) lesioned marmoset (Callithrix jacchus). Thirteen marmosets were trained on a battery of behavioural tasks that were conducted at regular intervals before and after surgery. All received unilateral 6-OHDA lesions to the medial forebrain bundle. Seven animals were then given an additional N-methyl-D-aspartate lesion of the ipsilateral subthalamic nucleus, whereas the remaining six animals received a variety of control or sham lesions to the nucleus. The 6-OHDA lesions induced a strong ipsilateral bias in head position; mild-moderate ipsilateral rotation spontaneously and after injection of saline or amphetamine; and contralateral rotation after injection of apomorphine. Hemineglect was evident as delayed initiation of reaches on the contralateral side on the staircase reaching task. Additional subthalamic lesions significantly reversed the bias in head position from ipsilateral to contralateral and decreased neglect as evidenced by improved latencies to initiate reaching on the contralateral side at the staircase. However, deficits in skilled movements persisted in the subthalamic nucleus lesion group in that they did not complete the staircase task any faster than the control group and remained impaired on another task which required reaching into tubes. These behavioural effects demonstrate that excitotoxic lesioning of the subthalamic nucleus can ameliorate some, but not all, parkinsonian-like deficits in the unilateral 6-OHDA lesioned marmoset. PMID- 9749731 TI - Localization of a G protein Gi2 in the cilia of rat ependyma, oviduct and trachea. AB - In previous studies, the localization of a pertussis toxin-sensitive G protein was demonstrated in ependymal cilia, but the identification of the subtype of G protein was inconsistent. To clarify this issue, we studied the localization of Goalpha, Gi1alpha, Gi3alpha and Gi2alpha in the ciliated ependymal cells and in the cilia of some other tissues of rats using specific antibodies. The cilia of the ependymal cells that line the ventricular cavity of the brain were intensely immunoreactive for Gi2alpha, but not for Goalpha, Gi1alpha or Gi3alpha. Immunoblot analysis demonstrated higher levels of Gi2alpha in the ependymal cilia rich pellet than in the motor area of the parietal cortex. At the ultrastructural level, the immunoreactivity specific for Gi2alpha was found predominantly in the cilia, but rarely in the microvilli or the basal bodies of ependymal cells. In cross-sections, the immunoreactivity specific for Gi2alpha was observed only in cell membranes, in particular, in the inner electron-dense leaflet of the trilaminar structure. In addition to that in the ependymal cilia, such specific localization of Gi2alpha was observed in the motile cilia in other tissues, including the oviduct and trachea. By contrast, the stereocilia in the ductus deferens were not immunopositive for Gi2alpha. These findings suggest that Gi2 might play an important role in the signal transduction in ciliary membrane associated function(s) of the ependymal cells, oviduct and trachea. PMID- 9749732 TI - Increased flexibility and selectivity in spatial learning of transgenic mice ectopically expressing the neural cell adhesion molecule L1 in astrocytes. AB - The expression of the neural cell adhesion molecule L1 is altered by neuronal activity and promotes neurite outgrowth in vitro. To study the effects of L1 on learning and synaptic plasticity, transgenic mice have been created which express L1 ectopically in glial fibrillary acidic protein (GFAP) expressing astrocytes. Ninety mice, including GFAP-L1-transgenic mice from two genetic backgrounds and their littermates, were tested for swimming navigation learning in the Morris water maze according to a standardized protocol. While learning the position of an invisible target platform and also relearning its position after relocation, GFAP-L1-transgenic mice spent a greater fraction of their swim time in the target quadrant. Moreover, they showed a more rapid improvement of escape performance during the first day of training. Factor analysis revealed that this difference in swimming pattern could not be explained by non-cognitive factors. Factor analysis also revealed that, during a probe trial, the GFAP-L1-transgenic mice spent comparatively less time in the old target quadrant than predicted by the increased searching they had shown during acquisition learning. Hence, ectopic expression of L1 by astrocytes in mice appears to be linked to a factor which increases behavioural flexibility and selectivity while learning and relearning, but concomitantly may lead to a relative reduction of spatial retention. PMID- 9749733 TI - Gamma frequency oscillation in the hippocampus of the rat: intracellular analysis in vivo. AB - Gamma frequency field oscillations reflect synchronized synaptic potentials in neuronal populations within the approximately 10-40 ms range. The generation of gamma activity in the hippocampus was investigated by intracellular recording from principal cells and basket cells in urethane anaesthetized rats. The recorded neurones were verified by intracellular injection of biocytin. Gamma frequency field oscillations were nested within the slower theta waves. The phase and amplitude of intracellular gamma were voltage dependent with an almost complete phase reversal at Cl- equilibrium potential in pyramidal cells. Basket cells fired at gamma frequency and were phase-locked to the same phase of the gamma oscillation as pyramidal cells. Current-induced depolarization coupled with synaptically induced inhibition resulted in gamma frequency discharge (30-80 Hz) of pyramidal cells without accommodation. These observations suggest that at least part of the gamma frequency field oscillation reflects rhythmic hyperpolarization of principal cells, brought about by the rhythmically discharging basket neurones. Resonant properties of pyramidal cells might facilitate network synchrony in the gamma frequency range. PMID- 9749734 TI - Dexterity in adult monkeys following early lesion of the motor cortical hand area: the role of cortex adjacent to the lesion. AB - Infant monkeys were subjected to unilateral lesions of the motor cortex (mainly its hand representation). After maturation, they showed normal use of the contralateral hand for global grip movements. However, as compared with the ipsilateral hand, precision grip tasks requiring relatively independent finger movements were performed with less dexterity, particularly if adjustments of the wrist position were necessary. The purpose of this study was to investigate mechanisms which may be responsible for the rather well, although not complete, preservation of manipulative behaviour of these adult monkeys. To this end, the hand representations were mapped bilaterally with intracortical microstimulation in the mature monkeys, and the dexterity of both hands assessed quantitatively in a precision grip task. The behavioural effects of reversible inactivations of the primary (M1) and supplementary (SMA) motor cortical areas were then tested. The following were found. (i) The hand contralateral to the lesion exhibited subtle but significant dexterity deficits, as compared with the ipsilateral hand; the deficit was essentially for complex movements requiring dissociation of the thumb index finger pinch from the other digits, involving also an arm rotation. (ii) Reversible inactivation of the M1 hand representation in the intact hemisphere dramatically impaired dexterity of the opposite hand without affecting the ipsilateral hand (contralateral to the early lesion). (iii) A relatively complete hand representation was found to occupy a new territory, medial to the old lesion. (iv) The role of this new displaced representation was crucial for the preserved dexterity of the opposite hand, as evidenced by its functional inactivation. In contrast, inactivation of both SMA cortices did not interfere with the manipulative behaviour. It is thus concluded that the preserved functional capacity of manipulations with the hand opposite the early lesion can be essentially attributed to a cortical reorganization around the old lesion. Under the present experimental conditions, contributions from either the SMA or the intact M1 appear not to be crucial. PMID- 9749735 TI - Compartments within human primary auditory cortex: evidence from cytochrome oxidase and acetylcholinesterase staining. AB - The human primary auditory area (AI) corresponds to granular cortex located on Heschl's gyrus. We studied its pattern of cytochrome oxidase and acetylcholinesterase activity in 10 normal human hemispheres. In cytochrome oxidase-stained coronal sections layer IV was prominent by its dark staining. The overall staining intensity varied along the medio-lateral extent of Al; a 2.0-2.5 mm-wide antero-posterior dark band was present at mid-AI. In acetylcholinesterase stained coronal sections a dark antero-posterior band appeared at the same location, corresponding to the highly granular part of Al. In cytochrome-oxidase stained tangential sections of flattened Al, approximately 500-microm thick alternating dark and light cytochrome oxidase stripes were present in layers III and IV. These stripes were perpendicular to the dark band. Comparison with tonotopic maps of human Al obtained by activation studies suggests that the cytochrome oxidase and acetylcholinesterase dark band is most likely parallel to isofrequency lines and may correspond to the representation of frequencies critical for speech comprehension. The narrow stripes may be related to particular binaural or ampliotopic domains, whose presence is suggested by evidence from electrophysiological recordings in cat Al and from magnetoencephalographic studies in humans. PMID- 9749736 TI - Demarcation of prosencephalic regions by CD15-positive radial glia. AB - A subpopulation of radial glial cells has been identified in the mouse prosencephalon during the second half of embryonic development. This subpopulation, specified by the putative cell adhesion molecule CD15 (LeX, FAL), is arranged in a segmented pattern within the telencephalon and diencephalon. Glial processes, spanning the prosencephalic wall, first appear at E10.5 and remain clearly visible until E19, when staining of discrete nuclei begins to appear. Registration of the correspondence between ventricular and pial surfaces, however, is still possible due to the persistence of individual CD15-positive fibres. These can be traced even when the initial simple linear (radial) orientation between ventricular and pial surfaces becomes complicated and distorted. After birth, CD15 immunoreactivity is distributed in a mosaic pattern in the forebrain. Because radial glial cells provide a scaffolding system for postmitotic neurones, the pattern of CD15-positive fibres in the embryonic prosencephalon may also demarcate future discrete regions of the postnatal brain. PMID- 9749737 TI - Influence of automatic word reading on motor control. AB - We investigated the possible influence of automatic word reading on processes of visuo-motor transformation. Six subjects were required to reach and grasp a rod on whose visible face the word 'long' or 'short' was printed. Word reading was not explicitly required. In order to induce subjects to visually analyse the object trial by trial, object position and size were randomly varied during the experimental session. The kinematics of the reaching component was affected by word presentation. Peak acceleration, peak velocity, and peak deceleration of arm were higher for the word 'long' with respect to the word 'short'. That is, during the initial movement phase subjects automatically associated the meaning of the word with the distance to be covered and activated a motor program for a farther and/or nearer object position. During the final movement phase, subjects modified the braking forces (deceleration) in order to correct the initial error. No effect of the words on the grasp component was observed. These results suggest a possible influence of cognitive functions on motor control and seem to contrast with the notion that the analyses executed in the ventral and dorsal cortical visual streams are different and independent. PMID- 9749738 TI - Anticonvulsant properties of BIBP3226, a non-peptide selective antagonist at neuropeptide Y Y1 receptors. AB - Several lines of evidence indicate that neuropeptide Y (NPY)-mediated neurotransmission in the hippocampus is altered by limbic seizures. The functional consequences of this change are still unresolved and clearly depend on the type of NPY receptors involved. We have investigated the role of NPY Y1 receptor subtypes, which are enriched in the dentate area of the hippocampus, on EEG seizures induced by a local injection of 0.04 microg kainic acid in rats. Intrahippocampal administration of 10 microg BIBP3226 (N2- (diphenylacetyl)-N-[(4 hydroxyphenyl)methyl]D-arginamide), a non-peptide selective antagonist at the NPY Y1 receptors, increased threefold on average (P < 0.01) the time to onset of seizures and reduced the number of seizures and the total time in seizures three- and fourfold, respectively (P < 0.01). Its inactive S-enantiomer BIBP3435 was ineffective on seizure activity. One microgram [Leu31,Pro34]NPY, an agonist at Y1 receptors, did not modify per se the EEG sequelae induced by kainic acid but it antagonized the anticonvulsant effect of BIBP3226. These results indicate that NPY Y1 receptors in the hippocampus are involved in epileptic phenomena and suggest that selective Y1 receptor antagonists may be of value for attenuating limbic seizures. PMID- 9749740 TI - Responses of human primary sensorimotor and supplementary motor areas to internally triggered unilateral and simultaneous bilateral one-digit movements. A high-resolution EEG study. AB - We modelled the responses of human primary sensorimotor areas and supplementary motor area to simple, self-initiated unilateral and simultaneous bilateral middle finger movements using a novel high-resolution electroencephalography technology. The results support the view that these cortical motor areas are involved in parallel and present similar activity in the preparation, initiation, and execution of the contralateral and bilateral movements. Furthermore, the left primary sensorimotor area (dominant hemisphere) appears to be activated more than the right primary sensorimotor area during the preparation and performance of the ipsilateral movements. PMID- 9749739 TI - Blockade of neuropsin, a serine protease, ameliorates kindling epilepsy. AB - The behavioural and electrographical abnormalities associated with seizures in epileptic (kindled) mice correspond with those of human epilepsy. In kindled mice, neuropsin was markedly increased in the hippocampus and cerebral cortices. A single intraventricular injection of monoclonal antibodies specific to neuropsin reduced or eliminated the epileptic pattern noted on electroencephalograms and, as a result markedly inhibited the progression of kindling. Therefore, neuropsin appears to be a key protein controlling pathogenic events in the hippocampus, and thus neuropsin inhibitors might be useful for treatment of epilepsy. PMID- 9749741 TI - Deafferentation up-regulates the expression of the mGlu1a metabotropic glutamate receptor protein in the olfactory bulb. AB - Chemical lesion of olfactory neuroepitheium induced an up-regulation of the mGlu1a metabotropic glutamate receptor protein in the olfactory bulb, as shown by Western blot analysis. At 2 days after the lesion, the increase in the receptor protein was associated with an increase in mGlu1a mRNA levels; in contrast, at longer times after the lesion (16 days), mRNA levels were reduced in spite of the high expression of the receptor protein, perhaps as a result of product inhibition of mGlu1 gene expression. Immunohistochemical analysis showed that the increase in mGlu1a induced by olfactory denervation was confined to the glomeruli, which occupy the external portion of the olfactory bulb. Within these structures, mGlu1a receptors are mainly localized on the distal dendrites of mitral cells, which are innervated by the glutamatergic axons of the olfactory nerve. These results demonstrate that the expression of mGlu1a receptors is up regulated in response to glutamatergic deafferentation, supporting a role for this particular receptor subtype in the physiology of synaptic transmission. PMID- 9749742 TI - A caspase inhibitor blocks ischaemia-induced delayed neuronal death in the gerbil. AB - Caspases play a critical role in the cell death machinery in various cell types. Here we investigated the involvement of caspases in the delayed neuronal death after transient global forebrain ischaemia in the gerbil. Intrahippocampal injection of benzyloxycarbonyl-Asp-CH2-dichlorobenzene (zD), an irreversible inhibitor of caspases, saved hippocampal CA1 neurones from chromatin condensation and DNA fragmentation at post-ischaemia day 4, and these neurones maintained normal morphology at day 8 post-insult. Intrahippocampal injection of interleukin 1beta (IL-1beta) after ischaemic insults did not influence the neuroprotective effect of zD, suggesting that the neuroprotective effect does not depend on the inhibition of mature IL-1beta production. Animals that received zD-injection showed significant improvement in step-through and step-down passive avoidance learning at post-ischaemia days 4 and 5, suggesting that neural functions were preserved in these animals. At post-ischaemia day 4, the cleavage of poly(ADP ribose)polymerase was observed, and this cleavage was almost completely suppressed in zD-injected hippocampus, suggesting involvement of caspase-3 and caspase-3-like caspase in the delayed neuronal death. Our findings indicate that caspases play important roles in the delayed neuronal death after transient global forebrain ischaemia in the gerbil, and suggest that ischaemia-induced brain damage can be blocked by caspase inhibitors. PMID- 9749743 TI - Long-lasting increased pain sensitivity in rat following exposure to heroin for the first time. AB - Acute dependence, defined as a precipitation of somatic signs by an antagonist, may occur after a single administration of an opiate drug. Because hyperalgesia is a consistent sign of the withdrawal syndrome, we tested the effectiveness of heroin, an opiate used by addicts, to induce pain facilitation even after a first exposure to the drug. In opiate-naive rats, subcutaneous injection of heroin induced analgesia followed by allodynia, a decrease in pain threshold. This latter phenomenon was observed in the absence of noxious stimuli and lasted several days. An N-methyl-D-aspartate (NMDA) receptor antagonist, MK-801 prevented such long-lasting allodynia. These results suggest that allodynia is an early sign reflecting neural plasticity associated with the development of dependence. PMID- 9749744 TI - Seizure prediction by non-linear time series analysis of brain electrical activity. AB - Brain electrical activity of 16 patients with temporal lobe epilepsy, recorded intracranially during seizure-free intervals as well as during transitions to the seizure state, was analysed using methods derived from the theory of non-linear dynamics. Long-lasting and marked changes towards low-dimensional system states were found to occur specifically up to 25 min prior to epileptic seizures and allow to predict the occurrence of individual seizures in time. These findings reflect a continuous increase in the degree of synchronicity, and thus open a window for the study of mechanisms generating seizures in humans. This offers new possibilities for therapeutic interventions. PMID- 9749745 TI - Pattern formation in the cerebellum of murine embryonic stem cell chimeras. AB - The cerebellar cortex is subdivided into an elaborate, stereotyped array of transverse zones and parasagittal stripes. It has been speculated that (i) all Purkinje cells derive from 10 to 20 precursors allocated early in embryogenesis and (ii) that pattern formation is based on cell lineage restriction in the founder pool. These hypotheses have been tested by clonal analysis of embryonic stem cell chimeras. Neither speculation is supported: the analysis suggests that Purkinje cells derive from a founder population of > 102 precursors, and that neither cerebellar transverse developmental boundaries nor parasagittal stripes have a clonal origin. We conclude that early lineage restriction plays no role in cerebellar pattern formation. PMID- 9749746 TI - Temporal limits of spatial working memory in humans. AB - An essential feature attributed to working memory is the labile and transient nature of its representations. Using an oculomotor task, we examined the stability of spatial working memory in 16 normal human subjects. Eye movements towards remembered spatial cues (memory-guided saccades) were electro oculographically recorded after memorization delays that varied unpredictably between 0.5 and 30s. A peaked time-course of saccadic targeting errors, with maximal errors around 20s delay, was found, showing that delay-dependent decay of spatial information in working memory occurs, but is time-limited and reverts significantly beyond delays of about 20s. These data (i) indicate temporal limits of spatial working memory and (ii) provide the first behavioural evidence for the existence of two parallely generated mental representations of space that successively control memory-guided behaviour in humans. PMID- 9749747 TI - Basic and acidic fibroblast growth factors protect spinal motor neurones in vivo after experimental spinal cord injury. AB - We studied the effect of a single focal injection of recombinant basic (FGF2) or acidic (FGF1) fibroblast growth factor on the survival of spinal motor neurones at 24 h after a standardized spinal cord contusion injury (SCI) in the rat. Both FGF2 and FGF1 (3 microg), microinjected into the injury site at 5 min post-injury (p.i.), protected at least two functionally important classes of spinal motor neurones, autonomic preganglionic neurones in the intermediolateral (IML) column and somatic motor neurones in the ventral horn (VH). Moreover, there was enhanced choline acetyltransferase (ChAT) immunoreactivity in surviving VH and IML neurones, suggesting an improved functional status. Thus, neurotrophic factors such as FGF2 and FGF1 may contribute to an overall strategy to treat acute SCI and improve recovery of function. PMID- 9749748 TI - Functional magnetic resonance imaging of a human auditory cortex area involved in foreground-background decomposition. AB - Auditory foreground-background decomposition is a pattern recognition process which combines simultaneous and sequential grouping in complex sound sequences. Using functional magnetic resonance imaging with reduced scanner noise and stimulation through a new type of earphones, we investigated the possibility that this process activates topographically distinct areas of human auditory cortex. A basic matching-to-sample task with variable tones (sequential grouping) caused significant activity in three separate landmark-related territories on the supratemporal plane. A similar task in the presence of a strongly masking acoustic background pattern to challenge simultaneous grouping led to the distinction of the subterritory in which foreground signal-related or task related signal properties were exclusively seen. In contrast to the remainder of territories the level of activity and the periodicity of the signal time-course was resistant to the masking influence of the background. This suggests that auditory foreground-background decomposition involves a specialized non-primary auditory cortex field. Generally, the findings demonstrate functional parcellation of auditory cortex for which the evidence in humans, in contrast to other primates, is only indirect to date. PMID- 9749749 TI - Effects of elevated levels of nerve growth factor on the septohippocampal system in transgenic mice. AB - Elevating target-derived levels of nerve growth factor (NGF) in peripheral organs of postnatal mammals is known to enhance the survival of postganglionic sympathetic neurons and to promote the terminal arborization of sympathetic axons within such NGF-rich target tissues. Although increasing levels of NGF in the central nervous system can ameliorate cholinergic function of damaged and aged neurons of the medial septum, it remains undetermined whether the postnatal development of this neuronal population and their projections that innervate the hippocampus are likewise affected by elevated levels of target-derived NGF. To address this question, the cholinergic septohippocampal pathway was examined in adult transgenic mice which display elevated levels of NGF protein production in the dorsal hippocampus during postnatal development. Adult transgenic mice possessed a cholinergic population of septal neurons approximately 15% larger than that seen in age-matched control animals. Despite increased numbers of cholinergic septal neurons, as well as elevated levels of hippocampal NGF, the density of cholinergic septal axons in the outer molecular layer of the hippocampal dentate gyrus of adult transgenic animals was comparable with that found in wild-type controls. These results reveal that elevating levels of target derived NGF during postnatal development can increase the population size of the cholinergic septal neurons but does not alter their pattern of afferent innervation in the hippocampus of adult mice. PMID- 9749750 TI - Differential increases in chromogranins, but not synapsin I, in cortical neurons following spreading depression: implications for functional roles and transmitter peptide release. AB - Experimental damage of cerebral cortex induces a slow-moving depolarization and subsequent depression of activity called cortical spreading depression (CSD) which is associated with various ionic, metabolic and genomic changes. Chromogranins are a family of water-soluble acidic proteins with a widespread distribution in secretory, large dense-core vesicles of neurons. We have earlier reported that secretogranin II (SgII) mRNA is increased in cerebral cortex hours after a unilateral craniotomy which would have induced CSD. To investigate further the regulation of chromogranin systems and the nature of genomic and biochemical changes produced by CSD, this study examined the temporal changes in chromogranin A (CgA), chromogranin B (CgB) and SgII mRNAs and CgB and SgII immunoreactivity (IR) in cerebral cortex and hippocampus following unilateral KCl induced CSD. For comparison, the levels of mRNA for synapsin I, a protein present in small synaptic vesicles was also examined. Rats were killed at various times after 10 min or 2 h of CSD and levels of chromogranins mRNAs were determined by semiquantitative in situ hybridization histochemistry, while changes in corresponding peptide products were detected by immunohistochemistry. CSD increased both SgII and CgB mRNA levels in ipsilateral cortex--levels of SgII mRNA were significantly (P < 0.01) increased at 1-6 h after CSD (165-225% of levels in contralateral cortex), but were not significantly above control values at later time points. Increased expression of CgB mRNA was delayed and prolonged compared with SgII and was significantly (P < 0.05) increased between 3 and 24 h (120-145%) after CSD, peaked at 2 days (180%), and was still elevated at 1 week (130%) compared with contralateral cortex. No alteration in CgA mRNA was observed in the ipsilateral cortex of the same animals across the entire time-course except for an increase in piriform cortex at 1-2 days. In contrast, levels of synapsin I mRNA in affected cortex were identical to those in contralateral cortex and cortex in sham-operated rats, at all times after CSD. Levels of chromogranin (SN-IR and PE-11-IR) were also increased in ipsilateral cortex following CSD. A strong increase in SN-IR in neuronal cell bodies and fibres was observed at 12 h and a moderate increase in PE-11-IR was observed 24-72 h after CSD. These results demonstrate that chromogranin transcripts and gene products are differentially regulated by neuronal depolarization/depression occurring during CSD and suggest that these chromogranin proteins may have differing functional roles in peptide transmitter release and distinct effects on neuronal function in rat brain. PMID- 9749751 TI - Increase in syntaxin 1B and glutamate release in mossy fibre terminals following induction of LTP in the dentate gyrus: a candidate molecular mechanism underlying transsynaptic plasticity. AB - A growing body of evidence suggests that modulation of certain proteins of the exocytotic machinery is, in part, involved in the biochemical changes that underlie long-term synaptic plasticity. We have previously shown that the induction of long-term potentiation (LTP) at perforant path to dentate granule cell synapses in the rat hippocampus induces changes in the mRNA levels of syntaxin 1B and synapsin I, known to be involved in neurotransmitter release. Immunohistochemical staining suggested that concomitant changes in these proteins occurred at mossy fibre synapses, downstream of those synapses at which LTP was induced, leading us to postulate that such a mechanism might underlie a form of transsynaptic plasticity. Here we have used a specific mossy-fibre synaptosome preparation to quantify levels of proteins and measure, using a chemiluminescent glutamate assay, depolarization-induced glutamate release from these synaptosomes after induction of LTP in the dentate gyrus in vivo. We show that 5 h after the induction of LTP, there is an increase in the protein levels of syntaxin 1B and, although to a lesser extent, the synapsins I and II, associated with an increase in depolarization-induced release of glutamate within these terminals. Increases in both the protein levels and glutamate release were not observed when dentate gyrus LTP was blocked by an NMDA receptor antagonist. From these results we propose a molecular mechanism for the propagation of synaptic plasticity through hippocampal circuits. PMID- 9749752 TI - Piracetam facilitates long-term memory for a passive avoidance task in chicks through a mechanism that requires a brain corticosteroid action. AB - We investigated the effects of piracetam, a nootropic, on learning and memory formation for a passive avoidance task in day-old chicks. To test for the possible cognitive-enhancing properties of piracetam, a weak learning version of this task--whereby chicks maintain a memory to avoid pecking at a bead coated in a diluted aversant for up to 10 h--was used. Post-training (5, 30 or 60 min), but not pretraining, injections of piracetam (10 or 50 mg/kg, i.p.) increased recall for the task when the chicks were tested 24 h later. Because previous studies showed that long-term memory for the passive avoidance task is dependent upon a brain corticosteroid action, and because the efficacy of piracetam-like compounds is also modulated by corticosteroids, we tested whether the facilitating effect of piracetam was dependent upon a corticosteroid action through specific brain receptors (mineralocorticoid receptor and glucocorticoid receptor). First, increased plasma levels of corticosterone were found 5 min after piracetam injection. In addition, intracerebral administration of antagonists for each receptor type (RU28318, for mineralocorticoid receptors, and RU38486 for glucocorticoid receptors; i.c.) given before the nootropic inhibited the facilitative effect of piracetam on memory consolidation. These results give further support to a modulatory action of piracetam on the mechanisms involved in long-term memory formation through a neural action that, in this learning model, requires the activation of the two types of intracellular corticosteroid receptors. PMID- 9749753 TI - Promoter analysis of the neuronal nicotinic acetylcholine receptor alpha4 gene: methylation and expression of the transgene. AB - Neuronal nicotinic acetylcholine receptor (nAChR) subunit genes compose a family of genes. The major isoform of nAChR in the brain is made up of the alpha4 and beta2 subunits and possesses a high affinity for nicotine. To investigate the mechanisms of the regulation of the nAChR alpha4 gene expression in mouse, its genomic DNA was cloned and characterized. The transcription initiation site was mapped by primer extension and RNase protection experiments and localized at about 254 bp upstream of the translation initiation site. The 5' flanking region of this gene did not have typical TATA box but GC-rich sequences were found around the initiation site. Methylation analysis of this region revealed that genomic DNAs from liver and muscle are partially methylated, whereas little methylation was observed in genomic DNA from brain. To characterize the cis acting elements driving cell-specific expression of the alpha4 subunit gene, we produced lines of transgenic mice which carry a series of fragments of the alpha4 gene fused with bacterial lacZ as a reporter gene. An 11.5-kb DNA fragment containing 9 kb of the region upstream of the transcription initiation site and the first intron was found to confer an expression pattern which coincides rather well with the endogenous gene expression pattern at early embryonic stages, suggesting that the elements necessary for the onset of alpha4 gene expression are located in this region. A DNA fragment containing the 1.8-kb upstream sequence and the first intron drove expression of lacZ in a limited subset of alpha4 expressing cells, whereas the 1.8-kb upstream sequence alone did not elicit any significant expression. These results show that both upstream and intronic sequences are important for cell-specific expression of the nAChR alpha4 gene. PMID- 9749755 TI - Importance of neurokinin-1 receptors in the nucleus tractus solitarii of mice for the integration of cardiac vagal inputs. AB - Unmyelinated vagal afferents from the heart terminate within the nucleus tractus solitarii (NTS) located in the dorsomedial medulla. The neurotransmitter and postsynaptic receptors mediating information from cardiac vagal receptors to the NTS are unknown. This study determined the effects of neurokinin-1 (NK1) receptor blockade on: (i) the reflex response evoked following aortic root injection of either veratridine (1-3 microg/kg) or bradykinin (80-300 ng/kg) to stimulate cardiac receptors in in vivo anaesthetized mice; and (ii) the evoked synaptic response of cardioreceptive NTS neurons following both intraleft-ventricular injection of veratridine or bradykinin, and electrical stimulation of the ipsilateral vagus nerve in an arterially perfused working heart-brainstem preparation of mouse. Administration of CP-99,994 (0.75-1.5 mg/kg i.v.), a specific NK1 antagonist, attenuated significantly the evoked reflex bradycardia and depressor response following cardiac receptor (n = 6), but not pulmonary chemoreflex stimulation in vivo. From extracellular recordings of cardioreceptive NTS neurons, CP-99,994 reduced reversibly the total number of evoked spikes, peak firing frequency and response duration evoked by intraventricular injections of veratridine (n = 5) or bradykinin (n = 5). The number of evoked action potentials following electrical stimulation of the vagus nerve was also reduced. In five whole cell recordings of NTS neurons, both the evoked depolarization following cardiac receptor stimulation, and the peak amplitude and duration of vagus nerve evoked EPSPs were reduced by CP-99,994; synaptic inputs from both peripheral chemoreceptors or pulmonary C-fibres were unaffected. These data support a selective involvement of NK1 receptors in the transmission of cardiac vagal afferent inputs to NTS neurons integrating cardiorespiratory information. PMID- 9749754 TI - Regional cerebral blood flow changes in human brain related to ipsilateral and contralateral complex hand movements--a PET study. AB - The purpose of this study was to investigate the cortical motor areas activated in relation to unilateral complex hand movements of either hand, and the motor area related to motor skill learning. Regional cerebral blood flow (rCBF) was measured in eight right-handed healthy male volunteers using positron emission tomography during a two-ball-rotation task using the right hand, the same task using the left hand and two control tasks. In the two-ball-rotation tasks, subjects were required to rotate the same two iron balls either with the right or left hand. In the control task, they were required to hold two balls in each hand without movement. The primary motor area, premotor area and cerebellum were activated bilaterally with each unilateral hand movement. In contrast, the supplementary motor area proper was activated only by contralateral hand movements. In addition, we found a positive correlation between the rCBF to the premotor area and the degree of improvement in skill during motor task training. The results indicate that complex hand movements are organized bilaterally in the primary motor areas, premotor areas and cerebellum, that functional asymmetry in the motor cortices is not evident during complex finger movements, and that the premotor area may play an important role in motor skill learning. PMID- 9749756 TI - Neuronal survival and calcium influx induced by basic fibroblast growth factor in chick ciliary ganglion neurons. AB - Basic fibroblast growth factor (bFGF/FGF2) exhibits widespread biological activities in the nervous system. However, little is known about the cascade of intracellular events that links the activation of its tyrosine kinase receptors to these effects. Here we report that, in ciliary ganglion neurons from chick embryo, this trophic factor significantly enhanced neuronal survival. The percentage of surviving neurons was reduced when intracellular calcium was chelated by adding a membrane-permeable BAPTA ester to the culture medium, while antagonists of L- and N-type voltage-dependent calcium channels were ineffective. The ionic signals in response to bFGF stimulation have been studied using cytofluorimetric and patch-clamp techniques. In single-cell Fura-2 measurements, bFGF elicited a long lasting rise of the cytosolic calcium concentration that was dependent on [Ca2+]o. In whole-cell experiments, we observed a reversible depolarization of the membrane resting potential and an inward cationic current. Single channel experiments, performed in the cell-attached configuration, provide evidence for the activation of two families of Ca2+-permeable cationic channels. Moreover, inositol 1,4,5-trisphosphate opens channels with similar properties, suggesting that this cytosolic messenger can be responsible for the calcium influx induced by bFGF. PMID- 9749757 TI - Postnatal expression of H1-receptor mRNA in the rat brain: correlation to L histidine decarboxylase expression and local upregulation in limbic seizures. AB - Histamine is implicated in the regulation of brain functions through three distinct receptors. Endogenous histamine in the brain is derived from mast cells and neurons, but the importance of these two pools during early postnatal development is still unknown. The expression of histamine H1-receptor in the rat brain was examined using in situ hybridization during postnatal development and in adults. For comparison, the expression of L-histidine decarboxylase (HDC) in the two pools was revealed. H1-receptor was evenly expressed throughout the brain on the first postnatal days, but resembled the adult, uneven pattern already on postnatal day 5 (P5). HDC was expressed in both mast cells and tuberomammillary neurons from birth until P5, after which the mast cell expression was no more detectable. In adult rat brain, high or moderate levels of H1-receptor expression were found in the hippocampus, zona incerta, medial amygdaloid nucleus and reticular thalamic nucleus. In most areas of the adult brain the expression of H1 receptor mRNA correlates well with binding data and histaminergic innervation. A notable exception is the hypothalamus, with high fibre density but moderate or low H1-receptor expression. Systemic kainic acid administration induced increased expression of H1-receptor mRNA in the caudate-putamen and dentate gyrus, whereas no change was seen in the hippocampal subfields CA1-CA3 or in the entorhinal cortex 6 h after kainic acid injections. This significant increase supports the concept that histaminergic transmission, through H1-receptor, is involved in the regulation of seizure activity in the brain. PMID- 9749758 TI - A peptide activator of endogenous tyrosine kinase enhances synaptic currents mediated by NMDA receptors. AB - N-methyl-D-aspartic acid (NMDA) receptor currents in cultured cells or expression systems are increased by the addition of purified tyrosine kinases. However, there is no direct demonstration of this effect at NMDA receptors in intact synapses of rat brain slices. Transmitters which might be used to activate tyrosine kinases in situ are unlikely to have a sufficiently selective action to allow a clear interpretation of their effects. Therefore, we used a phosphotyrosine-containing decapeptide which can be included in recording electrodes to activate postsynaptic src-family tyrosine kinases. This peptide enhanced NMDA responses in dissociated hippocampal CA1 neurons. These effects were not reproduced by a non-phosphorylated peptide or a scrambled-sequence phosphopeptide. The enhancement of NMDA responses was blocked by a tyrosine kinase inhibitor. In brain slices the phosphopeptide, but not control peptide, increased NMDA receptor-mediated synaptic current indicating that endogenous tyrosine kinase can upregulate the response of NMDA receptors at glutamatergic synapses in the hippocampus. PMID- 9749759 TI - The voltage-dependent conductances of rat neocortical layer I neurons. AB - Whole cell patch-clamp techniques were used to study voltage-dependent sodium (Na+), calcium (Ca2+), and potassium (K+) conductances in acutely isolated neurons from cortical layer I of adult rats. Layer I cells were identified by means of gamma-aminobutyric acid (GABA) immunocytochemistry. Positive stainings for the Ca2+-binding protein calretinin in a subset of cells, indicated the presence of Cajal-Retzius (C-R) cells. All investigated cells displayed a rather homogeneous profile of voltage-dependent membrane currents. A fast Na+ current activated at about -45 mV, was half-maximal steady-state inactivated at -66.6 mV, and recovery from inactivation followed a two-exponential process (tau1 = 8.4 ms and tau2 = 858.8 ms). Na+ currents declined rapidly with two voltage-dependent time constants, reaching baseline current after some tens of milliseconds. In a subset of cells (< 50%) a constant current level of < 65 pA remained at the end of a 90 ms step. A transient outward current (Ifast) activated approximately -40 mV, declined rapidly with a voltage-insensitive time constant (tau approximately 350 ms) and was relatively insensitive to tetraethylammonium (TEA, 20 mM). Ifast was separated into two components based on their sensitivity to 4-aminopyridine (4-AP): one was blocked by low concentrations (40 microM) and a second by high concentrations (6 mM). After elimination of Ifast by a conditioning prepulse (50 ms to -50 mV), a slow K+ current (I(KV)) could be studied in isolation. I(KV) was only moderately affected by 4-AP (6 mM), while TEA (20 mM) blocked most (> 80%) of the current. I(KV) activated at about -40 mV, declined monoexponentially in a voltage-dependent manner (tau approximately 850 ms at -30 mV), and revealed an incomplete steady-state inactivation. In addition to Ifast and I(KV), indications of a Ca2+-dependent outward current component were found. When Na+ currents, Ifast, and I(KV) were blocked by tetrodotoxin (TTX, 1 microM), 4-AP (6 mM) and TEA (20 mM) an inward current carried by Ca2+ was found. Ca2+ currents activated at depolarized potentials at about -30 mV, were completely blocked by 50 microM cadmium (Cd2+), were sensitive to verapamil (approximately 40% block by 10 microM), and were not affected by nickel (50 microM). During current clamp recordings, isolated layer I neurons displayed fast spiking behaviour with short action potentials (approximately 2 ms, measured at half maximal amplitude) of relative small amplitude (approximately 83 mV, measured from the action potential threshold). PMID- 9749760 TI - Modulation of big K+ channel activity by ryanodine receptors and L-type Ca2+ channels in neurons. AB - As metabotropic glutamate receptor type 1 (mGluR1) is known to couple L-type Ca2+ channels and ryanodine receptors (RyR, Chavis et al., 1996) in cerebellar granule cells, we examined if such a coupling could activate a Ca2+-sensitive K+ channel, the big K+ (BK) channel, in cultured cerebellar granule cells. We observed that (+/-)-1-amino-cyclopentane-trans-1,3-dicarboxylic acid (t-ACPD) and quisqualate (QA) stimulated the activity of BK channels. On the other hand, (2S, 3S, 4S) alpha-carboxycyclopropyl-glycine (L-CCG-I) and L-(+)-2-amino-4-phosphonobutyrate (L-AP4) had no effect on BK channels, indicating a specific activation by group I mGluRs. Group I mGluRs stimulation of the basal BK channel activity was mimicked by caffeine and both effects were blocked by ryanodine and nifedipine. Interestingly, carbachol stimulated BK channel activity but through a pertussis toxin (PTX)-sensitive pathway that was independent of L-type Ca2+ channel activity. Our report indicates that unlike the muscarinic receptors, group I mGluRs activate BK channels by mobilizing an additional pathway involving RyR and L-type Ca2+ channels. PMID- 9749761 TI - PET study of human voluntary saccadic eye movements in darkness: effect of task repetition on the activation pattern. AB - Using H2(15)O 3D Positron Emission Tomography (PET), regional cerebral blood flow (rCBF) was measured in six human subjects under two different conditions: at rest and while performing self-paced horizontal saccadic eye movements in darkness. These two conditions were repeated four times each. First, the comparison between the four saccadic and four resting conditions was investigated in a group and a single subject analysis. Saccades elicited bilateral rCBF increases in the medial part of the superior frontal gyrus (supplementary eye field), precentral gyrus (frontal eye field), superior parietal lobule, anterior medial part of the occipital lobe involving striate and extrastriate cortex (lingual gyrus and cuneus), and in the right inferior parietal lobule. At the subcortical level, activations were found in the left putamen. These results mainly replicate previous PET findings on saccadic control. Second, the interaction between the experimental conditions and their repetition was examined. When activations throughout repetition of the same saccadic task are compared, the supplementary eye fields show a progressive increase of activation. On the contrary, the activation in the cerebellum, left superior parietal lobule and left occipital cortex progressively decreases during the scanning session. Given the existence of such an interaction, the pattern of activations must be interpreted as a function of task repetition. This may be a factor explaining some apparent mismatch between different studies. PMID- 9749762 TI - Expression and presence of septal neurokinin-2 receptors controlling hippocampal acetylcholine release during sensory stimulation in rat. AB - We examined the expression and presence of NK2 receptors in the septal area of rat brain, and investigated their functional role in the regulation of the septohippocampal cholinergic system. Using reverse transcription-polymerase chain reaction (RT-PCR) analysis, we showed the presence of NK2 receptor mRNA expression in the septal area, and detected septal NK2 binding sites by using a fluorescent-tagged neurokinin A (NKA) derivative. In vivo microdialysis was employed to explore the functional role of NK2 receptors in the release of hippocampal acetylcholine evoked by tactile stimulation in freely moving rats. Two sessions of stroking of the neck and back of the rat for 30 min, at 90 min intervals, produced a marked and reproducible increase in hippocampal acetylcholine release. This effect was dose-dependently prevented by intraperitoneal administration of the two selective non-peptide tachykinin NK2 receptor antagonists SR144190 (0.03-0.3 mg/kg, i.p.) and SR48968 (0.3 and 1 mg/kg, i.p.), but not by the inactive enantiomer of SR48968 (SR48965, 1 mg/kg) nor by the two non-peptide NK1 receptor antagonists SR140333 (3 mg/kg, i.p.) and GR205171 (1 mg/kg, i.p.). Furthermore, the intraseptal application of SR144190 (10(-8) M) reduced the sensory response. Finally, intraseptal perfusion of neurokinin A (0.01-10 microM) in anaesthetized rats produced a concentration dependent increase in hippocampal acetylcholine release. The response to neurokinin A (0.1 microM) was prevented by SR144190 (0.03-0.3 mg/kg, i.p.) and SR48968 (0.3-1 mg/kg, i.p.). In conclusion, this study provides direct evidence for the role of endogenous NKA/substance P, through the activation of NK2 receptors, in regulating the septohippocampal cholinergic function. PMID- 9749763 TI - A population of cholinergic neurons is present in the macaque monkey thalamus. AB - Three new cholinergic markers were employed to study the cholinergic innervation in the thalamus of adult macaque monkeys. They were: two antibodies against choline acetyltransferase (ChAT), one polyclonal and one monoclonal; and a polyclonal antibody against the vesicular transporter of acetylcholine (VAChT), a powerful new marker that colocalizes with ChAT. This approach led to an unexpected finding: the three antibodies positively immunostained a population of neurons in the paracentral nucleus. The immunostained cells are confined to the dorsal region of this nucleus along its rostrocaudal extent. Measurement of the somatic areas of the immunostained neurons indicated that they correspond to a population of large neurons thought to be projection neurons. Because dorsal paracentral neurons are known to project to the dorsal striatum and specific cortical areas involved in visual and visuomotor mechanisms, these structures might be modulated by cholinergic thalamic neurons. PMID- 9749764 TI - Amelioration of spatial navigation and short-term memory deficits by grafts of foetal basal forebrain tissue placed into the hippocampus and cortex of rats with selective cholinergic lesions. AB - Impairments in learning and memory, induced by surgical or excitotoxic lesions of the septo-hippocampal or basalo-cortical pathways, can be ameliorated by grafts of cholinergic-rich foetal basal forebrain tissue into the hippocampus and/or neocortex. However, the effects of such grafts have been only partial, which may be due to the non-specific nature of the lesioning procedures used in these studies, known to destroy both cholinergic and non-cholinergic neuronal projections. In the present study, we have explored the effects of cholinergic rich grafts in rats subjected to selective cholinergic lesions, induced by intraventricular injections of the immunotoxin 192 IgG-saporin. This lesion, which selectively destroyed 85-95% of the cholinergic neurons in both the septal diagonal band and nucleus basalis, produced a long-lasting, substantial impairment in both the acquisition of spatial reference memory in the Morris water maze task and delay-dependent short-term memory performance, as seen in a delayed matching-to-position test. Foetal cholinergic grafts (but not control grafts of cerebellar tissue) implanted at multiple sites into both the hippocampus and fronto-parietal neocortex, bilaterally, completely reversed the acquisition deficit in place navigation in the water maze, to an extent that greatly exceeded that previously seen in animals with non-selective lesions. Most notably, however, the impairment in short-term memory was only partially and inconsistently affected, and only at the longest delay times. The morphological analysis, performed at about 7 months after transplantation, showed that the grafts had re-established a close to normal cholinergic innervation in the initially denervated cortical and hippocampal territories. It is proposed that the differential effects of cholinergic-rich transplants on different aspects of cognitive performance may define intrinsic limitations to the functional capacity of the ectopically placed grafts, which may be due to incomplete integration of the grafted cholinergic neurons into functional regulatory circuitries normally available to the basal forebrain cholinergic system. PMID- 9749765 TI - Serotonin inhibits synaptic glutamate currents in rat nucleus accumbens neurons via presynaptic 5-HT1B receptors. AB - Neurons in the nucleus accumbens septi in brain slices from adult male rats were studied with patch clamp recording in the whole-cell conformation. Cells filled with Lucifer Yellow were identified as medium spiny neurons. Electrical stimulation close to the recorded cell evoked excitatory and inhibitory synaptic currents. In the presence of picrotoxin or bicuculline, stimulation at a holding potential of -90 mV evoked an inward excitatory current that was blocked by 6 cyano-7-nitroquinoxaline-2,3-dione (CNQX, 10 microM), identifying it as an excitatory postsynaptic current (EPSC) mediated by glutamate acting at AMPA/kainate receptors. Serotonin (5-hydroxytryptamine, 5-HT; 3-100 microM in the bath) decreased the EPSC in about 90% of the cells. The action of 5-HT was mimicked by N-(3-trifluoromethylphenyl)-piperazine HCl (TFMPP), but not by (+/-) 8-hydroxydipropylaminotetralin (8-OH-DPAT) or (+/-)-2,5-dimethoxy-4 iodoamphetamine HCl (DOI). The 5-HT effect was antagonized by pindolol or cyanopindolol, but not by spiperone, ketanserin or tropisetron. Taken together, these results indicate that 5-HT acts at 5-HT1B receptors. The effect of 5-HT was potentiated by cocaine (0.3-3 microM) or the selective serotonin reuptake inhibitor citalopram. Miniature synaptic currents recorded in the presence of tetrodotoxin were inhibited by CNQX, identifying them as spontaneous miniature EPSCs. 5-HT reduced the frequency of these miniature EPSCs without affecting their amplitude, which indicates a presynaptic site of action. This presynaptic inhibition by 5-HT might be involved in the behavioural effects of cocaine. PMID- 9749766 TI - Effects of chronic alcohol consumption or Diazepam administration on item recognition and temporal ordering in a spatial working memory task in mice. AB - This study was aimed at determining the effects of either Diazepam administration or chronic alcohol consumption (CAC) on spatial memory measured by concurrent discriminations in an eight arm radial maze using mice as subjects. Two different protocols involving a non-matching rule were used to evaluate either temporal order (recurrent items) or item recognition (non-recurrent items). Results showed that both Diazepam administration and CAC produced a memory deficit which was primarily observed in the temporal task, whereas item recognition was spared. These data show that Diazepam and CAC produced similar memory impairments. Thus, our study stressed the potential importance of the GABA/BDZ dysfunction in the production of organic amnesia of alcoholic origin. The overall analysis of the data suggests that both CAC and Diazepam injections would impair forms of memory sustained by automatic or incidental learning. PMID- 9749767 TI - Increase of preprotachykinin mRNA and substance P immunoreactivity in spared dorsal root ganglion neurons following partial sciatic nerve injury. AB - Complete sciatic nerve injury reduces substance P (SP) expression in primary sensory neurons of the L4 and L5 dorsal root ganglia (DRG), due to loss of target derived nerve growth factor (NGF). Partial nerve injury spares a proportion of DRG neurons, whose axons lie in the partially degenerating nerve, and are exposed to elevated NGF levels from Schwann and other endoneurial cells involved in Wallerian degeneration. To test the hypothesis that SP is elevated in spared DRG neurons following partial nerve injury, we compared the effects of complete sciatic nerve transection (CSNT) with those of two types of partial injury, partial sciatic nerve transection (PSNT) and chronic constriction injury (CCI). As expected, a CSNT profoundly decreased SP expression at 4 and 14 days postinjury, but after PSNT and CCI the levels of preprotachykinin (PPT) mRNA, assessed by in situ hybridization, and the SP immunoreactivity (SP-IR) of the L4 and L5 DRGs did not decrease, nor did dorsal horn SP-IR decrease. Using retrograde labelling with fluorogold to identify spared DRG neurons, we found that the proportion of these neurons expressing SP-IR 14 days after injury was much higher than in neurons of normal DRGs. Further, the highest levels of SP-IR in individual neurons were detected in ipsilateral L4 and L5 DRG neurons after PSNT and CCI. We conclude that partial sciatic nerve injury elevates SP levels in spared DRG neurons. This phenomenon might be involved in the development of neuropathic pain, which commonly follows partial nerve injury. PMID- 9749768 TI - Cytokine-induced changes in the ability of astrocytes to support migration of oligodendrocyte precursors and axon growth. AB - Repair of demyelination in the CNS requires that oligodendrocyte precursors (OPs) migrate, divide and then myelinate. Repair of axon damage requires axonal regeneration. Limited remyelination and axon regeneration occurs soon after injury, but usually ceases in a few days. In vivo and in vitro experiments have shown that astrocytic environments are not very permissive for migration of OPs or for axonal re-growth. Yet remyelination and axon sprouting early after injury occurs in association with astrocytes, while later astrocytes can exclude remyelination and prevent axon regeneration. A large and changing cast of cytokines are released following CNS injury, so we investigated whether some of these alone or in combination can affect the ability of astrocytes to support migration of OPs and neuritic outgrowth. Interleukin (IL) 1alpha, tumour necrosis factor alpha, transforming growth factor (TGF) beta, basic fibroblast growth factor (bFGF), platelet-derived growth factor and epidermal growth factor alone exerted little or no effect on migration of OPs on astrocytes, whereas interferon (IFN) gamma was inhibitory. The combination of IL-1alpha + bFGF was found to be pro-migratory, and this effect could be neutralized by TGFbeta. We also examined neuritic outgrowth from dorsal root ganglion explants in three-dimensional astrocyte cultures treated with cytokines and found that IL-1alpha + bFGF greatly increased axon outgrowth and that this effect could be blocked by TGFbeta and IFNgamma. All these effects were absent or much smaller when OP migration or axon growth was tested on laminin, so the main effect of the cytokines was via astrocytes. The cytokine effects did not correlate with expression on astrocytes of laminin, fibronectin, tenascin, chondroitin sulphate proteoglycan, N-cadherin, polysialyated NCAM (PSA-NCAM), tissue plasminogen activator (tPA) or urokinase (uPA). PMID- 9749769 TI - Mechanisms of spontaneous cytosolic Ca2+ transients in differentiated human neuronal cells. AB - We have studied Ca2+ homeostasis in a unique model of human neurons, the NT2N cell, which differentiates from a human teratocarcinoma cell line, NTera2/C1.D1 by retinoic acid treatment. When perifused with Krebs-HEPES buffer containing 2.5 mM CaCl2, fura-2 loaded NT2N cells produced spontaneous cytosolic Ca2+ oscillations, or Ca2+ transients. These cytosolic Ca2+ transients were not blocked by antagonists of glutamate (6-cyano-7-nitroquinoxaline-2,3-dione and D( )-2-amino-5-phosphonopentanoic acid) or muscarinic (atropine) receptors. Omission of extracellular Ca2+ completely abolished Ca2+ oscillations and decreased the average Ca2+ level from 106 +/- 14 nM to 59 +/- 8 nM. Addition of the L-type Ca2+ channel blocker nifedipine (1 or 10 microM) or of the N-type inhibitor omega conotoxin GVIA (5 microM) significantly, although incompletely, suppressed Ca2+ oscillations, while omega-conotoxin MVIIC (5 microM), a selective antagonist of P and Q-channels, had no effect. Ni2+, at 100 microM, a concentration selective for T-type channels, did not inhibit Ca2+ transients. Non-specific blockage of Ca2+ channels by higher concentrations of Ni2+ (2-5 mM) or Co2+ (1 mM) abolished Ca2+ oscillations completely. The endoplasmic reticulum Ca2+-ATPase inhibitor, thapsigargin (1 microM), slightly decreased Ca2+ oscillation frequency, and induced a small transitory increase in the average cytosolic Ca2+ concentration. The mRNAs of L- (alpha1D subunit) and N-type (alpha1B subunit) Ca2+ channel were present in NT2N cells, while that of a T-type Ca2+ channel (alpha1-subunit) was not present in the NT2N cells as shown by reverse transcription-polymerase chain reaction. In conclusion, NT2N neuronal cells generate cytosolic Ca2+ oscillations mainly by influx of extracellular Ca2+ through multiple channels, which include L and N-type channels, and do not require activation of glutamate or muscarinic receptors. PMID- 9749770 TI - Action sequencing is impaired in D1A-deficient mutant mice. AB - The role of dopamine in the production of behaviour is multifarious in that it can influence different aspects of movement (e.g. movement initiation, sensorimotor integration, and movement sequencing). A characteristic of the dopamine system which seems to be critical for the expression of this diverse influence is its varied receptor population. Previous studies have shown that specific receptor subtype activation leads to specific behavioural responses or alterations of selective aspects of movement. It is known that one of the important influences of dopamine includes sequential co-ordination of 'syntactic' patterns of grooming movements because moderate loss of the dopaminergic nigrostriatal projections specifically disrupts these patterns without affecting grooming actions in a general fashion (Berridge, K.C. Psychobiology, 15, 336, 1989). The specific receptors of the dopamine family which play a key part in this co-ordination of movement sequences is not known. In the present study, we examined the serial order of particular syntactic sequences or chains of grooming actions in mice lacking D1A receptors to explore the relationship between this receptor subtype and movement sequencing. Mutant mice had shorter grooming bouts and a disruption of the organization of sequential patterns compared with wild type littermate controls. Sequential disruption was reflected in the failure of D1A mutants to follow the syntactic pattern of grooming to completion. This sequential disruption deficit appeared to be specific, as mutant mice initiated more syntactic chains than wild-type controls even though they were less likely to complete them. These results support the hypothesis that D1A receptor activation plays a part in the sequencing of natural action. This conclusion has important implications for the understanding of the functional heterogeneity of dopamine receptor subtypes and of the aetiology of symptoms observed in patients with basal ganglia disease. PMID- 9749771 TI - Action potential broadening induced by lithium may cause a presynaptic enhancement of excitatory synaptic transmission in neonatal rat hippocampus. AB - Lithium enhances excitatory synaptic transmission in CA1 pyramidal cells, but the mechanisms remain unclear. The present study demonstrates that lithium enhances the N-methyl-D-aspartate (NMDA) and alpha-amino-3-hydroxy-5-methyl-isoxazole propionic acid (AMPA) receptor-mediated components of the excitatory postsynaptic current (EPSC). Lithium decreased the magnitude of paired-pulse facilitation and presented an inverse correlation between the lithium-induced enhancement of synaptic transmission and initial paired-pulse facilitation, which is consistent with a presynaptic mode of action. The enhancement of synaptic strength is likely to act, at least in part, by increasing the amplitude of the presynaptic Ca2+ transient. One mechanism which could account for this change of the presynaptic Ca2+ transient is an increase in the duration of the action potential. We investigated action potential in hippocampal pyramidal neurons and found that lithium (0.5-6 mM) increased the half-amplitude duration and reduced the rate of repolarization, whereas the rate of depolarization remained similar. To find out whether the lithium synaptic effects might be explained by spike broadening, we investigated the field recording of the excitatory postsynaptic potential (EPSP) in hippocampal slices and found three lines of evidence. First, the prolongation of the presynaptic action potential with 4-aminopyridine and tetraethylammonium blocked or reduced the synaptic effects of lithium. Second, the lithium-induced synaptic enhancement was modulated when presynaptic Ca2+ influx was varied by changing the external Ca2+ concentration. Finally, both effects, the synaptic transmission increment and the action potential broadening, were independent of inositol depletion. These results suggest that lithium enhances synaptic transmission in the hippocampus via a presynaptic site of action: the mechanism underlying the potentiating effect may be attributable to an increased Ca2+ influx consequent to the broadening effect of lithium on the action potential. PMID- 9749772 TI - Immunorecognition, ultrastructure and phosphorylation status of astrocytic gap junctions and connexin43 in rat brain after cerebral focal ischaemia. AB - Gap junctions between astrocytes support a functional syncytium that is thought to play an important role in neural homeostasis. In order to investigate regulation of this syncytium and of connexin43 (Cx43), a principal astrocytic gap junction protein, we determined the sequelae of gap junction and Cx43 disposition in a rat cerebral focal ischaemia model with various ischaemia/reperfusion times using sequence-specific anti-Cx43 antibodies (designated 13-8300, 18A, 16A and 71 0700) that exhibit differential recognition of Cx43, perhaps reflecting functional aspects of gap junctions. Antibody 13-8300 specifically detects only an unphosphorylated form of Cx43 in both Western blots and tissue sections. In hypothalamus after brief (15 min) ischaemic injury, Cx43 at intact gap junctions undergoes dephosphorylation, accompanied by reduced epitope recognition by antibodies 16A and 71-0700. Tissue examined 24 h after reperfusion showed that these effects were reversible. Astrocytic gap junction internalization occurring 1 h after ischaemia was accompanied by decreased immunodetection with 13-8300. At this time, gap junctions were absent in the ischaemic core, coinciding with a loss of Cx43 recognition with 18A and 13-8300, but elevated labelling of internalized Cx43 with 16A and 71-0700. Unphosphorylated Cx43 persisted at intact gap junctions confined to a thin corridor at the ischaemic penumbra which contained presumptive apoptotic cell profiles. Similar results were obtained in ischaemic striatum and cerebral cortex, though with a delayed time course that depended on the severity of the ischaemic insult. These results demonstrate that astrocytic Cx43 epitope masking, dephosphorylation and cellular redistribution occur after ischaemic brain injury, proceed as a temporally and spatially ordered sequence of events and culminate in differential patterns of Cx43 modification and sequestration at the lesion centre and periphery. These observations suggest an attempt by astrocytes in the vicinity of injury to remodel the junctional syncytium according to altered tissue homeostatic requirements. PMID- 9749773 TI - Cellular prion protein localization in rodent and primate brain. AB - The presence of an abnormal, protease-resistant form of the prion protein (PrP) is the hallmark of various forms of transmissible spongiform encephalopathies (TSE) which can affect a number of mammalian species, including humans. The normal, cellular form of this protein, PrPc, while abundant in brain is also present in many tissues and a number of species. In order to address the unresolved question of the precise localization of normal cerebral PrPc, we used a free-floating immunohistochemistry procedure to localize the protein at both the light and the electron microscopic levels in the brain of three TSE-sensitive species: hamster, macaque and humans. This method shows that PrPc is abundant in synaptic terminal fields in olfactory bulb, limbic-associated structures and in the striato-nigral complex, whereas many other regions of the hamster brain are essentially devoid of immunoreactivity. With the striking exception of the olfactory nerve, in which axons are continually growing throughout life, PrPc is not abundant in fibre pathways. PrPc distribution in the primate hippocampus and cortex is very similar to the distribution observed in hamster. PrPc was present at synaptic profiles as shown by immunoelectron microscopy, but was not detectable in neuronal perikaryon either by light or electron microscopy. Our results show that PrPc is abundant in a number of brain structures known for ongoing plasticity, and are consistent with the hypothesis that the protein also plays a role in synaptic function. PMID- 9749774 TI - Short-lasting exposure to one odour decreases general reactivity in the olfactory bulb of adult rats. AB - We investigated in adult rats whether a relatively short exposure to a novel odour can lead to changes in reactivity of olfactory bulb principal neurons. Naive rats were exposed to isoamyl acetate for 20 min per day either for 6 consecutive days or for a single 20-min exposure. Control group was non-exposed. Under anaesthesia, responsiveness of each recorded single mitral/tufted cell was tested towards isoamyl acetate and four other odours. Results show that the proportion of responding cells in the exposed groups decreased drastically when compared to controls. In the two experimental groups recorded 24 h following the last exposure, mitral/tufted cells show a significant decrease in the number of excitatory responses. In parallel, the number of non-responsive cells increased by at least a fourfold factor. This decrease in reactivity was not selective towards the odour used during the exposure but concerned any of the five test odours presented during recordings. Finally, this lower responsiveness was long lasting as it was still observed 10 days after the end of the last exposure. This preliminary study points out the importance of even limited sensory experience in neural representation of odours. PMID- 9749775 TI - Induction of long-term potentiation at spinal synapses by noxious stimulation or nerve injury. AB - Use-dependent long-term potentiation of synaptic strength (LTP) is an intensively studied model for learning and memory in vertebrates. Induction of LTP critically depends on the stimulation parameters of presynaptic fibres with synchronous high frequency bursts being most effective at many central synapses. It is, however, not known whether naturally occurring discharge patterns may induce LTP and whether LTP has any biological function in sensory systems. Here we have investigated the LTP of excitatory synaptic transmission between primary afferent C-fibres, many of which are nociceptors, and neurons in rat superficial spinal dorsal horn. LTP that lasted for 4-6 h could not only be induced by electrical stimulation of sural nerve but also by natural stimulation of heat-, mechano- or chemosensitive nociceptors in the skin or by acute nerve injury. Maintenance of LTP was not affected when afferent nerves were cut 1 h or 5 min after noxious skin stimulation, indicating that an ongoing afferent barrage is not required. Natural noxious stimuli induced LTP in animals which were spinalized but were ineffective in intact animals. Thus, induction of LTP is suppressed by tonically active supraspinal descending systems. We conclude that the natural non synchronized discharge patterns that are evoked by noxious stimulation may induce LTP and that this new form of LTP may be an underlying mechanism of afferent induced hyperalgesia. PMID- 9749776 TI - Differential patterns of semaphorin expression in the developing rat brain. AB - Semaphorins are a large family of cell-surface and secreted proteins that have been shown to function as chemorepellents or inhibitors of growth cones of peripheral neurons, yet little is known about their role in patterning central pathways. In order to examine whether semaphorins may be involved in guiding the formation of the reciprocal thalamocortical connections in the rat, we have analysed the spatial and temporal expression of five recently identified rodent semaphorins (semB, C, D, F and G) using in situ hybridization. Transcripts of all five genes were present throughout the period examined (E15-P7) and displayed highly specific spatiotemporal distributions. We have based our discussion of putative semaphorin effects on their known functions as chemorepellents and found their spatiotemporal expression patterns compatible with such a role in several developmental events. Specifically, semaphorins are in the position to: (i) prevent neurite extension into the ventricular neuroepithelium throughout the brain; (ii) confer non-permissive properties to the embryonic cortical plate, hence regulating the radial invasion of corticopetal afferents; (iii) confine axonal extension to the intermediate zone and subplate; (iv) maintain the fasciculated state of thalamocortical and corticothalamic axons, and prevent them from branching while they grow through the striatum; and (v) restrict the terminal arborizations of thalamic afferents to layer IV. The evidence that different semaphorin genes are often co-expressed further suggests that the various molecules might interact in synergistic ways. Taken together, our results support the hypothesis that semaphorins could act as guidance signals in the development of the thalamocortical projections and suggest that innervation specificity is achieved through the combined action of multiple guidance cues. Furthermore, these data provide a basis for the design of functional assays and the study of mice carrying knockouts in specific semaphorin genes. PMID- 9749777 TI - Intraventricular galanin modulates a 5-HT1A receptor-mediated behavioural response in the rat. AB - The present studies have examined whether the neuropeptide galanin can modulate brain serotoninergic (5-HT) neurotransmission in vivo and, particularly, 5-HT1A receptor-mediated transmission. For that purpose, we studied the ability of galanin (given bilaterally into the lateral ventricle, i.c.v.) to modify the impairment of passive avoidance retention induced by the selective 5-HT1A agonist 8-hydroxy-2-(di-n-propyloamino)tetralin (8-OH-DPAT) when injected prior to training. This impairment appears to be mainly related to activation of 5-HT1A receptors in the CNS. Galanin dose-dependently (significant at 3.0 nmol/rat) attenuated the passive avoidance impairment (examined 24 h after training) induced by the 0.2 mg/kg dose of 8-OH-DPAT. This 8-OH-DPAT dose produced signs of the 5-HT syndrome indicating a postsynaptic 5-HT1A receptor activation. Furthermore, both the impairment of passive avoidance and the 5-HT syndrome were completely blocked by the 5-HT1A receptor antagonist WAY 100635 (0.1 mg/kg). Galanin (0.3 or 3.0 nmol) or WAY 100635 (0.1 mg/kg) failed by themselves to affect passive avoidance retention. 8-OH-DPAT given at a low dose 0.03 mg/kg, which presumably stimulates somatodendritic 5-HT1A autoreceptors in vivo, did not alter passive avoidance retention or induce any visually detectable signs of the 5-HT syndrome. Galanin (0.3 or 3.0 nmol) given i.c.v. in combination with the 0.03 mg/kg dose of 8-OH-DPAT, did not modify passive avoidance. The immunohistochemical study of the distribution of i.c.v. administered galanin (10 min after infusion) showed a strong diffuse labelling in the periventricular zone (100-200 microm) of the lateral ventricle. Furthermore, in the dorsal and ventral hippocampus galanin-immunoreactive nerve cells appeared both in the dentate gyrus and the CA1, CA2 and CA3 layers of the hippocampus. In the septum only endogenous fibres could be seen while in the caudal amygdala also galanin-immunoreactive nerve cells were visualized far away from the labelled periventricular zone. At the level of the dorsal raphe nucleus a thin periventricular zone of galanin immunoreactivity was seen but no labelling of cells. These results suggest that galanin can modulate postsynaptic 5-HT1A receptor transmission in vivo in discrete cell populations in forebrain regions such as the dorsal and ventral hippocampus and parts of the amygdala. The indication that galanin administered intracerebroventrically may be taken up in certain populations of nerve terminals in the periventricular zone for retrograde transport suggests that this peptide may also affect intracellular events. PMID- 9749778 TI - Basolateral amygdala stimulation evokes glutamate receptor-dependent dopamine efflux in the nucleus accumbens of the anaesthetized rat. AB - Afferents from the basolateral amygdala and dopamine projections from the ventral tegmental area to the nucleus accumbens have both been implicated in reward related processes. The present study used in vivo chronoamperometry with stearate graphite paste electrodes in urethane-anaesthetized rats to determine how basolateral amygdala efferents to the nucleus accumbens synaptically regulate dopamine efflux. Repetitive-pulse (20 Hz for 10 s) electrical stimulation of the basolateral amygdala evoked a complex pattern of changes in monitored dopamine oxidation currents in the nucleus accumbens related to dopamine efflux. These changes were characterized by an initial increase that was time-locked to stimulation, a secondary decrease below baseline, followed by a prolonged increase in the dopamine signal above baseline. The effects of burst-patterned stimulation (100 Hz, 5 pulses/burst, 1-s interburst interval, 40 s) of the basolateral amygdala on the basal accumbens dopamine signal were similar to those evoked by 20 Hz stimulation, with the lack of a secondary suppressive component. Infusions of the ionotropic glutamate receptor antagonists (+/-)-2-amino-5 phosphonopentanoic acid (APV) or 6,7-dinitroquinoxaline-2,3-dione (DNQX) into the nucleus accumbens dose-dependently blocked or attenuated the initial and prolonged increases in the dopamine signal following 20 Hz or burst-patterned basolateral amygdala stimulation. Infusions of the metabotropic glutamate receptor antagonist (+)-alpha-methyl-4-carboxyphenylglycine selectively blocked the intermediate suppressive effect of 20 Hz basolateral amygdala stimulation on dopamine oxidation currents. Blockade of glutamate receptors or inhibition of dopamine neuronal activity via infusions of either APV + DNQX, lidocaine or gamma hydroxybutyric acid, respectively, into the ventral tegmental area did not effect the pattern of changes in the accumbens dopamine signal evoked by basolateral amygdala stimulation. These data suggest that the glutamatergic basolateral amygdala inputs to nucleus accumbens dopamine terminals synaptically facilitate or depress dopamine efflux, and these effects are independent of dopamine neuronal firing activity. Moreover, these results imply that changes in nucleus accumbens dopamine levels following presentation of reward-related stimuli may be mediated, in part, by the basolateral amygdala. PMID- 9749779 TI - Potentiation of murine astrocyte antioxidant defence by bcl-2: protection in part reflects elevated glutathione levels. AB - Overexpression of the proto-oncogene bcl-2 has been shown to protect a variety of cell types from oxidative and non-oxidative injury, blocking apoptotic and necrotic types of cell death. Retroviral vectors were used to stably overexpress bcl-2 in primary murine astrocyte cultures with more than 95% efficiency. Compared to beta-galactosidase-expressing and uninfected control cells, bcl-2 overexpressing astrocytes suffered < 40% injury after 24 h glucose deprivation, while controls were essentially completely injured. After exposure to 0.2 mM hydrogen peroxide, the bcl-2 overexpressing astrocytes suffered < 40% the injury seen in controls. In contrast, when the cultures were injured by combined oxygen glucose deprivation, no difference in the extent or time course of injury was found between cells overexpressing bcl-2 and those expressing beta-galactosidase. To investigate one possible mechanism of bcl-2 protection, we measured the levels of glutathione and three antioxidant enzymes. Astrocytes overexpressing bcl-2 had elevated glutathione levels (130-200%), increased superoxide dismutase (170%) and glutathione peroxidase (140%) activities compared with beta-galactosidase expressing controls. Bcl-2 overexpressing astrocytes suffered less lipid peroxidation after glucose deprivation, as assessed by cis-parinaric acid fluorescence, and demonstrated more rapid removal of hydrogen peroxide from the medium. When glutathione levels were decreased 80% by pretreatment with buthionine sulfoximine, the extent of protection from glucose deprivation of bcl 2 overexpressing cells was decreased by about half. Increased antioxidant defence contributes to protection from glucose deprivation in bcl-2 overexpressing astrocytes. PMID- 9749780 TI - Layer-specific dendritic regression of pyramidal cells with ageing in the human prefrontal cortex. AB - The dendritic field of pyramidal neurons in cortical layers IIIc and V of the prefrontal cortex in ageing humans was studied. The three-dimensional branching pattern of the basilar dendrites of Golgi-Cox impregnated neurons was analysed in the middle frontal gyrus (areas 9 and 46) in eight subjects between the ages of 49 and 90 years, all without a neurological or psychiatric disorder. The results revealed a significant regression of the layer V dendritic pattern with increasing age, but the layer IIIc neurons did not show any age-related changes. Together with our earlier data on the postnatal development of the same cell types in the prefrontal cortex, we hypothesize that the layer V neurons in the prefrontal cortex start to regress from the fifth decade onwards, in contrast to the layer IIIc neurons which remain stable from puberty on. We conclude that pyramidal cells in layer IIIc and V in a similar cortical region undergo a differential ageing effect. PMID- 9749781 TI - Differentiation of rat hypothalamic dopaminergic neurons is stimulated in vitro by target cells: the melanotrophs. AB - We have investigated in vitro the influence of pituitary intermediate lobe melanotrophs on the differentiation of their afferent hypothalamic dopaminergic neurons. The presence of melanotrophs in primary cultures of foetal hypothalamic neurons induces an increase of the number of dopaminergic neurons (while the total neuronal population remains unchanged) and induces a stimulation of their neuritic outgrowth. These effects are mediated by diffusible factors since they are reproduced by application of conditioned medium issued from co-cultures with intermediate lobe cells from newborn rats. Moreover, by immunoneutralization of alpha-melanocyte-stimulating hormone (alphaMSH) in the co-culture or conditioned medium, or by application of the peptide itself, we demonstrate that the neuritotrophic effect on dopaminergic neurons is mediated by alphaMSH, the main secretory product of melanotrophs, whereas the inductive effect on the number of dopaminergic neurons is attributable to another diffusible neurotrophic factor(s) present in foetal, but not adult, adenohypophysis. Similar effects are observed on cultures of newborn hypothalamic neurons. However, at this stage of neuronal development, alphaMSH also increases the number of dopaminergic neurons, which could be due to a change of neuronal receptivity. We show that the neuritotrophic influence of alphaMSH is restricted to the dopaminergic neurons connected to the melanotrophs, and that in addition, these neurons systematically co-express the tyrosine hydroxylase and glutamate decarboxylase as the neurons innervating the melanotrophs in situ. These findings indicate that the differentiation of dopaminergic hypothalamic neurons is influenced by the target cells, melanotrophs, and that this trophic influence implicates alphaMSH. PMID- 9749782 TI - Endogenous nerve growth factor regulates the sensitivity of nociceptors in the adult rat. AB - Nerve growth factor (NGF) has a well characterized role in the development of the nervous system and there is evidence that it interacts with nociceptive primary afferent fibres. Here we applied a synthetic tyrosine kinase A IgG (trkA-IgG) fusion molecule for 10-12 days to the innervation territory of the purely cutaneous saphenous nerve in order to bind, and thereby neutralize endogenous NGF in adult rats. Using neurophysiological analysis of 152 nociceptors we now show that sequestration of NGF results in specific changes of their receptive field properties. The percentage of nociceptors responding to heat dropped significantly from a normal 57% to 32%. This was accompanied by a rightward shift and a reduced slope of the stimulus response function relating the intracutaneous temperature to the neural response. The number of nociceptors responding to application of bradykinin was also significantly reduced from a normal of 28% to 8%. In contrast, the threshold for mechanical stimuli and the response to suprathreshold stimuli remained unaltered, as did the percentage of nociceptors responding to noxious cold. The reduced sensitivity of primary afferent nociceptors was accompanied by a reduction in the innervation density of the epidermis by 44% as assessed with quantitative immunocytochemical analysis of the panaxonal marker PGP 9.5. This demonstrates that endogenous NGF in the adult specifically modulates the terminal arborization of unmyelinated fibres and the sensitivity of primary afferent nociceptors to thermal and chemical stimuli in vivo. PMID- 9749783 TI - Small sensory neurons in the rat dorsal root ganglia express functional NK-1 tachykinin receptor. AB - The tachykinins substance P (SP) and neurokinin A, released by the C-type primary afferent fibre terminals of the small dorsal root ganglion (DRG) neurons, play important roles in spinal nociception. By means of non-radioactive in situ hybridization and whole-cell recording, we showed that the small rat DRG neurons also express the NK-1 tachykinin receptor. In situ hybridization demonstrated that the positive neurons in rat DRG sections were mainly small cells (85.9%) with diameters less than 25 microm. The remaining positive neurons (14.1%) were cells with medium diameters between 26 and 40 microm. No positive large neurons (diameters > 40 microm) were observed. Expression in small DRG neurons (diameter < 21 microm) was confirmed by in situ hybridization of isolated cells, which were demonstrated to express NK-1 receptor mRNA at a very high frequency (> 90% of small DRG neurons) and therefore were subjected to whole-cell recording. In 57 of 61 cells recorded, SP or the selective NK-1 receptor agonist [Sar9, Met(O2)11]SP (Sar-SP, 1 or 2 microM) produced a delayed vibrating inward current (50-300 nA) with a long duration of 0.5-2 h. These currents were blocked by co-application of the NK-1 receptor antagonist L-668, 169 (1 microM), but were not affected by the NK-2 antagonist L-659, 877 (2 microM). Both current-clamp recording and cell attached single-channel recording demonstrated that the long-lasting response was due to the opening of a channel with an inward current. Employment of non-Ca2+ and Ca2+ + choline solutions revealed that this channel might be a Ca2+ permeable, non-selective cation channel. The prolonged NK-1 tachykinin response exhibited extreme desensitization. This work suggests that presynaptic NK-1 autoreceptors may be present on the primary afferent terminals in the spinal cord, where they could contribute to the chronic pain and hyperalgesia. PMID- 9749784 TI - Chemosensitivity of nociceptive, mechanosensitive afferent nerve fibres in the guinea-pig ureter. AB - The mechanosensitivity and chemosensitivity of afferent fibres were investigated in an in vitro preparation of the guinea-pig ureter. Electrophysiological recordings were obtained from 5 U-1 (low mechanical threshold, contraction sensitive) and 74 U-2 units (high threshold). U-2 units had significant higher levels of spontaneous activity, lower conduction velocities, higher mechanical thresholds (U-1: 7 mmHg; U-2: 39 mmHg), less pronounced phasic responses and longer latencies in the response to distensions than the U-1 units. For chemical stimulation, guinea-pig urine (> 800 mosmol/L), bradykinin and capsaicin were applied intraluminally. The responses of U-1 units mainly corresponded to the contractions induced by the chemical stimulation. The vast majority of the U-2 units were excited by urine, bradykinin (threshold: 0.1-1 microM) and capsaicin (threshold: 0.03-0.3 microM). The responses to urine could be mimicked by high concentrations of potassium ions (> 200 mM), but not by an equiosmolar solution of NaCl, urea and mannitol. Chemical stimulation could also result in a transient sensitization of the U-2 units to mechanical stimuli. In the anaesthetized guinea pig, pseudo-affective responses could be evoked by ureteric distension (threshold: 30-60 mmHg) and serosal application of capsaicin. Intraluminal application of urine in vivo did not evoke any reactions, suggesting that the responses of the U-2 units to urine might be due to an impaired barrier function of the urothelium in vitro. The data are in agreement with the hypothesis that U 2 units are visceral polymodal nociceptors. Since the U-1 units were also able to encode at least noxious mechanical stimuli, their involvement in visceral nociception cannot be excluded. PMID- 9749785 TI - Modulation of the voltage-gated sodium current in rat striatal neurons by DARPP 32, an inhibitor of protein phosphatase. AB - DARPP-32 is a cyclic adenosine monophosphate-regulated inhibitor of protein phosphatase 1, highly enriched in striatonigral neurons. Stimulation of dopamine D1 receptors increases phosphorylation of DARPP-32, whereas glutamate acting on N methyl-D-aspartate receptors induces its dephosphorylation. Yet, to date, there is little direct evidence for the function of DARPP-32 in striatal neurons. Using a whole cell patch-clamp technique, we have studied the role of DARPP-32 in the regulation of voltage-gated sodium channels in rat striatal neurons maintained in primary culture. Injection of phospho-DARPP-32, but not of the unphosphorylated form, reduced the sodium current amplitude. This effect was similar to those induced by okadaic acid, with which there was no additivity and by tautomycin. Our results indicate that, in striatal neurons, sodium channels are under dynamic control by phosphorylation/dephosphorylation, and that phospho-DARPP-32 reduces sodium current by stabilizing a phosphorylated state of the channel or an associated regulatory protein. We propose that the DARPP-32-mediated modulation of sodium channels, via inhibition of phosphatase 1, contributes to the regulation of these channels by D1 receptors and other neurotransmitters which influence the state of phosphorylation of DARPP-32. PMID- 9749786 TI - Cytoskeletal and nuclear localization of myelin oligodendrocytic basic protein isoforms. AB - The recently described single copy myelin-associated oligodendrocytic basic protein (Mobp) gene is expressed exclusively in the central nervous system (CNS). The gene encodes a family of small highly basic polypeptides with predicted amino acid lengths of 69, 71, 81, 99 and 170, all of which share a 68 residue amino terminal. Here we report on the subcellular distribution of two of these polypeptides termed MOBP81 and MOBP170 in transiently transfected Cos7 cells using an antibody raised against a region common to all isoforms of MOBP. Additionally, we describe MOBP trafficking in cultured mouse spinal cord oligodendrocytes. Immunostaining for MOBP81 is intense in the perinuclear region and extends throughout the cytoplasm colocalizing with the microtubular cytoskeletal network. Consistent with this we demonstrate that MOBP partitions with the cytoskeletal fraction prepared from myelin. In contrast, although MOBP170 is present in the cytoplasm it does not colocalize with the cytoskeleton and displays a greater variation in distribution. In the majority of transfectants immunostaining is present throughout the karyoplasm but with increased intensity around the nucleolus. Within mouse primary oligodendrocytes endogenous MOBP is present in the cell body and processes colocalizing with the microtubular network. Immunoreactivity is not detectable in the nucleus in these mature oligodendrocytes. These significant differences in MOBP81 and MOBP170 protein kinesis coupled to different expression profiles of their respective message populations may be indicative of both myelin structural and cellular/regulatory functions, respectively, for these polypeptides. PMID- 9749787 TI - A role for slow NMDA receptor-mediated, intrinsic neuronal oscillations in the control of fast fictive swimming in Xenopus laevis larvae. AB - In larvae of the amphibian, Xenopus laevis, spinal neurons which are active during fictive swimming also display tetrodotoxin-resistant membrane potential oscillations following the coactivation of N-methyl-DL-aspartate (NMDA) and 5 hydroxytryptamine (serotonin or 5-HT) receptors (Scrymgeour-Wedderburn et al., 1997; Eur. J. Neurosci., 9, 1473-1482). The oscillations are slow (approximately 0.5 Hz) compared with swimming (approximately 7-35 Hz) raising doubt over their contribution to the cycle by cycle depolarizations occurring during swimming. We investigated an alternative: that the intrinsic oscillations modulate swimming activity over many consecutive cycles. Bath application of NMDA induced continuous fictive swimming that differed between embryonic and larval preparations. In 81% of larval preparations (n = 36), there was a slow (approximately every 2 s) rhythmic modulation of ventral root activity in which burst durations and intensities increased as cycle periods decreased. This pattern of activity was enhanced rather than abolished following blockade of glycine and gamma-aminobutyric acid (GABA) A receptors and presumably therefore resulted from a periodic increase in the excitation of motor neurons. To determine whether this slow rhythm resulted from intrinsic, 5-HT-dependent membrane potential oscillations, larvae were spinalized to prevent the release of 5-HT from brainstem raphe neurons. The resulting pattern of NMDA-induced activity lacked any slow modulation. The slow modulation could also be enhanced by the bath application of a 5-HT receptor agonist (5-carboxamidotryptamine) and abolished either by the addition of an antagonist (pindobind-5-HT1A) or by removal of magnesium ions, providing more direct evidence for a contribution of intrinsic oscillations. Thus, the 5-HT-dependent intrinsic oscillations modulate NMDA-induced swimming activity over several consecutive cycles. PMID- 9749788 TI - Serotonergic modulation of the responses to excitatory amino acids of rat dorsal horn neurons in vitro: implications for somatosensory transmission. AB - Serotonin (5-HT) is one of the major transmitters involved in supraspinal control of somatic sensation and nociception. The aim of the present study was to investigate if the 5-HT-induced modulation of sensory transmission in the dorsal horn could be due to regulation of neuronal responses to excitatory amino acids. Experiments were performed in an in vitro preparation of the young rat spinal cord. Responses to dorsal root stimulation (DR-EPSP) and to droplet application of N-methyl-D-aspartic acid (NMDA) and alpha-amino-2,3-dihydro-5-methyl-3-oxo-4 isoxazolepropanoic acid (AMPA) were obtained by means of intracellular recordings of dorsal horn neurons. Bath applications of 5-HT (50 microM) generally caused reductions in amplitude and integrated area of DR-EPSPs and of responses to NMDA but the responses to AMPA were unaltered. A linear correlation was found between the effects of 5-HT on the DR-EPSP and on the NMDA response measured as percentage change in amplitude (r2 = 0.45; P < or = 0.01) and integrated area (r2 = 0.77; P < or = 0.001). The NMDA receptor antagonist d-AP5 (50 microM) completely abolished NMDA responses and caused a depression of the DR-EPSP similar to that of 5-HT. The 5-HT1 receptor agonist 5-carboxamidotryptamine (5 CT; 1 microM) mimicked the depressant effects of 5-HT but had a stronger depressant action on the DR-EPSP than 5-HT. The depression of NMDA responses induced by 5-HT and 5-CT was tetrodotoxin (1 microM) resistant. It is concluded that 5-HT-induced depression of NMDA responses explains partially the depressant action of 5-HT on dorsal horn synaptic transmission activating a postsynaptic site sensitive to 5-CT. The possible activation of coadjuvant mechanisms is discussed. PMID- 9749789 TI - Differential expression of voltage-gated K+ and Ca2+ currents in bipolar cells in the zebrafish retinal slice. AB - Whole-cell voltage-gated currents were recorded from bipolar cells in the zebrafish retinal slice. Two physiological populations of bipolar cells were identified. In the first, depolarizing voltage steps elicited a rapidly activating A-current that reached peak amplitude < or = 5 ms of step onset. IA was antagonized by external tetraethylammonium or 4-aminopyridine, and by intracellular caesium. The second population expressed a delayed rectifying potassium current (IK) that reached peak amplitude > or = 10 ms after step onset and did not inactivate. IK was antagonized by internal caesium and external tetraethylammonium. Bipolar cells expressing IK also expressed a time-dependent h current at membrane potentials < -50 mV. Ih was sensitive to external caesium and barium, and was also reduced by Na+-free Ringer. In both groups, a calcium current (ICa) and a calcium-dependent potassium current (IK(Ca)) were identified. Depolarizing voltage steps > -50 mV activated ICa, which reached peak amplitude between -20 and -10 mV. ICa was eliminated in Ca+2-free Ringer and blocked by cadmium and cobalt, but not tetrodotoxin. In most cells, Ica was transient, activating rapidly at -50 mV. This current was antagonized by nickel. The remaining bipolar cells expressed a nifedipine-sensitive sustained current that activated between -40 and -30 mV, with both slower kinetics and smaller amplitude than transient ICa. IK(Ca) was elicited by membrane depolarizations > -20 mV. Bipolar cells in the zebrafish retinal slice preparation express an array of voltage-gated currents which contribute to non-linear I-V characteristics. The zebrafish retinal slice preparation is well-suited to patch clamp analyses of membrane mechanisms and provides a suitable model for studying genetic defects in visual system development. PMID- 9749790 TI - A phenytoin-sensitive cationic current participates in generating the afterdepolarization and burst afterdischarge in rat neocortical pyramidal cells. AB - We report here on the ionic mechanisms underlying the depolarizing afterpotential (DAP) in neocortical pyramidal cells, with special interest in those underlying the burst afterdischarge. Injections of short depolarizing current pulses under whole-cell current clamp with a CsCl-based internal medium generated, in most pyramidal cells, a single action potential with a plateau phase (plateau-AP), followed by a slowly decaying DAP both in the absence and presence of TTX. Under voltage-clamp, the same cells displayed a slow tail current (tail-I) at the offset of depolarization. When intracellular free Ca2+ was chelated with 10 mM BAPTA or when extracellular Ca2+ was replaced with equimolar Ba2+, neither the slow DAP nor the slow tail-I was observed. Extracellular application of Co2+ or Cd2+ reduced Ca2+ currents and the slow tail-I. Cation substitution experiments revealed that the channel generating the slow tail-I was permeable to K+ and Cs+ more than to Na+ (PK is approximately equal to PCs > PNa > PNMDG is approximately equal to PTEA). The cationic slow tail-I was not reduced by applying antagonists of the metabotropic glutamate receptor (MCPG, 1 mM) and the muscarinic receptor (atropine, 1-10 microM). Thus, the slow DAP was produced by activation of the cationic channel whose gating is solely dependent on [Ca2+]i. An increase in [K+]o from 3 to 6 or 9 mM enhanced the slow DAP, and resulted in a generation of burst afterdischarges. An anticonvulsant, phenytoin (PT; 1-10 microM) suppressed the slow DAP while enhancing the plateau-AP in the presence of TTX, most likely by blocking the cationic channel. PMID- 9749791 TI - Inositol 1,4,5-trisphosphate activates non-selective cation conductance via intracellular Ca2+ increase in isolated frog taste cells. AB - The effect of intracellular Ca2+ increase was analysed in isolated frog taste cells under the whole-cell patch clamp. External application of a Ca2+-ionophore, ionomycin (3 microM) induced the sustained inward current of -200+/-17 pA (mean +/- SE, n = 23) at -50 mV in taste cells. The ionomycin-induced response was observed in most of the cells exposed in the drug, but not when 10 mM BAPTA (1,2 bis (O-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid) was included in the pipette (eight cells). Steady-state I-V relationships of ionomycin-induced currents were almost linear and reversed at -8+/-1 mV (n = 23). The simultaneous removal of Na+ and Ca2+ from the external solution eliminated the response completely (three cells). Intracellular dialysis with 1 mM Ca2+ or 50 microM inositol 1,4,5-trisphosphate (IP3) in K+-internal solution also induced an inward current in the taste cells. The Ca2+-induced and IP3-induced responses were observed in 82% and 36% of the cells dialysed with the drugs, respectively. The Ca2+-induced and IP3-induced currents were inhibited by external Cd2+ (1-2 mM). The reversal potentials of the inward currents were -15+/-3 mV (n = 9) in Ca2+ dialysis and -11+/-3 mV (n = 13) in IP3 dialysis. The half-maximal Ca2+ concentration in the pipette to induce the inward current was approximately 170 microM. The results suggest that IP3 can depolarize the taste cell with mediation by intracellular Ca2+. PMID- 9749792 TI - Expression of the mitotic motor protein CHO1/MKLP1 in postmitotic neurons. AB - The kinesin-related motor protein CHO1/MKLP1 was initially thought to be expressed only in mitotic cells, where it presumably transports oppositely oriented microtubules relative to one another in the spindle mid-zone. We have recently shown that CHO1/MKLP1 is also expressed in cultured neuronal cells, where it is enriched in developing dendrites [Sharp et al. (1997a) J. Cell Biol., 138, 833-843]. The putative function of CHO1/MKLP1 in these postmitotic cells is to intercalate minus-end-distal microtubules among oppositely oriented microtubules within developing dendrites, thereby establishing their non-uniform microtubule polarity pattern. Here we used in situ hybridization to determine whether CHO1/MKLP1 is expressed in a variety of rodent neurons both in vivo and in vitro. These analyses revealed that CHO1/MKLP1 is expressed within various neuronal populations of the brain including those in the cerebral cortex, hippocampus, olfactory bulb and cerebellum. The messenger ribonucleic acid (mRNA) levels are high within these neurons well after the completion of their terminal mitotic division and throughout the development of their dendrites. After this, the levels decrease and are relatively low within the adult brain. Parallel analyses on developing hippocampal neurons in culture indicate that the levels of expression increase dramatically just prior to dendritic development, and then decrease somewhat after the dendrites have differentiated. Dorsal root ganglion neurons, which generate axons but not dendrites, express significantly lower levels of mRNA for CHO1/MKLP1 than hippocampal or sympathetic neurons. These results are consistent with the proposed role of CHO1/MKLP1 in establishing the dendritic microtubule array. PMID- 9749793 TI - Rewarding effects elicited by the microinjection of either AMPA or NMDA glutamatergic antagonists into the ventral tegmental area revealed by an intracranial self-administration paradigm in mice. AB - In order to study the functional role of the trans-synaptic neuronal interaction between glutamatergic afferents and mesolimbic dopaminergic neurons in internal reward processes, BALB/c male mice were unilaterally implanted with a guide cannula, the tip of which was positioned 1.5 mm above the ventral tegmental area (VTA). On each day of the following experimental period, a stainless steel injection cannula was inserted into the VTA in order to study the eventual self administration behaviour of either the competitive N-methyl-D-aspartate antagonist, D(-)-2-amino-7-phosphonoheptanoic acid (AP-7) or the alpha-amino-3 hydroxy-5-methyl-isoxazole-4-propionic acid antagonist, 6,7-dinitroquinoxaline 2,3-dione (DNQX) (3 ng/50 nL) using a spatial discrimination task in a Y maze. Mice rapidly discriminated between the arm enabling a microinjection of either of these glutamatergic antagonists and the neutral arm of the maze, and a robust self-administration of either of these compounds was observed from the first session of acquisition. These data provide strong evidence that the intra-VTA microinjection of either of these subclasses of glutamatergic antagonist produces an effect which is interpreted centrally by the experimental subjects as being highly rewarding. Once the self-administration response had been fully acquired by the experimental subjects, preinjection of the dopaminergic D2 antagonist, sulpiride (50 mg/kg i.p.), 30 min before the test, produced a rapid extinction of the self-administration response. This latter result demonstrates the dopaminergic D2 receptor dependence of this intra-VTA self-administration of both of these subclasses of glutamatergic antagonist. We conclude that the different glutamatergic afferent neuronal inputs to the VTA globally exert, in vivo, via the mediation of interposed endogenous GABAergic interneurons, a tonic trans synaptic inhibitory regulation of neuronal activity in the mesolimbic dopaminergic pathway and that this complex neuronal interaction in the VTA plays a significant functional part in the modulation of internal reward processes. PMID- 9749794 TI - Death-signalling cascade in mouse cerebellar granule neurons. AB - Molecular mechanisms of neuronal cell death are still largely unknown. In the present study, the signal transduction pathway of cell death in cerebellar granule neurons was examined by employing various death-preventative agents. When death was induced by the depletion of serum and a depolarizing level of potassium, transient increase in active c-Jun, mitochondrial membrane potential (deltapsi) loss, activation of caspase-3 (-like) proteases, and nuclear condensation and fragmentation were observed. The protein synthesis inhibitor cycloheximide blocked all these phenomena, whereas RNA synthesis inhibitor actinomycin-D, survival factor such as insulin-like growth factor-1, brain derived neurotrophic factor, high K+ (25 mM) and overproduced antiapoptotic protein Bcl-2, prevented deltapsi, loss, caspase activation, and nuclear change, but not an increase in active c-Jun. The caspase inhibitor z-Asp-CH2-DCB (carbobenzoxy-L-aspartyl-alpha-[(2,6-dichlorobenzoyl) oxy]methane) only inhibited activation of caspases and nuclear change. These results suggest that the death signal in cerebellar granule neurons is sequentially transduced in the order of c Jun activation, de novo RNA synthesis, mitochondrial deltapsi loss, activation of caspase-3 (-like) proteases and nuclear change. PMID- 9749795 TI - Hyperpolarizing after-potentials regulate generation of long-duration plateau depolarizations in rat paraventricular nucleus neurons. AB - Activation of N-methyl-D-aspartate (NMDA) receptors in a population of neurons of the paraventricular nucleus (PVN) results in long-duration plateau depolarizations during which the membrane rapidly depolarizes, reaching a stable plateau near -20 mV. These responses were observed in 29% of the Type II PVN neurons tested with 1 microM NMDA agonist (n = 84). The stable plateau phase is characterized by an increase in ionic conductance, from 1.19+/-0.11 nS to 5.24+/ 2.17 nS (n = 5). Bath application of tetrodotoxin (n = 4) or alternatively inclusion of QX-314 in the pipette solution (n = 3) prevented the generation of these events. The remaining cells tested (n = 56) also depolarized in response to NMDA agonist, but long duration plateau depolarizations were not observed. Previous evidence from hypothalamic cultures has demonstrated synaptically driven plateau potentials following the blockade of repolarizing conductances. Pharmacological blockade of the post-spike hyperpolarizing afterpotential with 4 aminopyridine (200 microM), in cells that did not generate plateaux, resulted in the observance of long duration plateau depolarizations in response to a subsequent application of NMDA agonist (n = 4). Our results demonstrate that this 4-aminopyridine-sensitive ionic conductance plays a critical role in determining whether a cell will depolarize for a prolonged duration in response to NMDA receptor activation. As a prolonged depolarization of the postsynaptic membrane and accompanying membrane permeability changes are essential for neurotoxicity, these findings provide evidence for a potential protective mechanism that depends solely on the ability of the cell, through its ionic conductances, to control imposed changes in membrane potential. PMID- 9749796 TI - Visual activity is required to maintain the phenotype of supragranular NPY neurons in rat area 17. AB - Visual activity governs the functional maturation of the mammalian visual cortex. We report here, that visual experience is required for stabilizing the phenotype of a subset of cortical interneurons. Neurons expressing neuropeptide Y mRNA (NPY neurons) display a transiently higher expression in the early postnatal visual areas 18a and 17 that is followed by a phenotype restriction during the second postnatal month: about 50% of the NPY neurons in supragranular and infragranular layers of area 18a, and in infragranular layers of area 17 gradually stop the NPY expression. In contrast, the expression remains unchanged in supragranular layers of area 17. Dark rearing rats from birth to up to 100 days does neither prevent the developmental onset of NPY mRNA expression, nor does it prevent the phenotype restriction from occurring. In contrast, in dark reared animals NPY neurons in supragranular layers of area 17 now also undergo a phenotype restriction. Returning animals to light after variable periods of darkness results in an upregulation of NPY mRNA expression selectively in neurons in supragranular layers of area 17. These neurons acquire a constitutive expression during the second postnatal month. This suggests that the phenotypic specification of a distinct subset of cortical interneurons is regulated by visual experience which thus influences on the maturation of the neurochemical architecture of area 17. PMID- 9749797 TI - Reduction of noradrenaline impairs attention and dopamine depletion slows responses in Parkinson's disease. AB - We investigated the role dopamine and noradrenaline in the modulation of attention in Parkinson's disease (PD) patients. We observed that PD patients with mild and moderate motor disability did not differ in their attentional accuracy in easy tests, but the severe PD group was slightly disrupted in a more arduous test of attention. Attentional accuracy was not affected by withdrawal of dopaminergic drugs in mild or severe PD patients. The movements of severe PD patients were slower, and withdrawal of dopaminergic drugs aggravated motor slowing more in severe PD patients. Clonidine (0.5 and 2 microg/kg) retarded accuracy of performance in the most difficult attention test in mild PD patients, but had no effect in the severe PD group. Clonidine had no effect on movement times. These data suggest that a defect in noradrenaline release may contribute to the impaired accuracy of attention in severe PD patients and that dopamine may be important for maintaining rapid motor responding. PMID- 9749798 TI - Basic fibroblast growth factor promotes the generation and differentiation of calretinin neurons in the rat cerebral cortex in vitro. AB - Calretinin-expressing neurons are some of the earliest postmitotic cells to appear in the developing cerebral cortex. Lineage studies have shown that the expression of this calcium-binding protein in cortical neurons is not genetically programmed and is likely to be induced by external factors. A number of studies have clearly shown that basic fibroblast growth factor (bFGF) and a number of neurotrophins promote the proliferation and differentiation of cortical progenitor cells to a particular lineage. Here, using a culture system of dissociated rat cortical cells, we found that brain-derived neurotrophic factor and neurotrophin-3 promoted the morphological differentiation of one of the calretinin-containing neuronal subpopulations, the Cajal-Retzius cells. Another subpopulation of calretinin-expressing cells of smaller size and bipolar form was generated when cultures were treated with bFGF. The progenitors of these neurons were stimulated by bFGF to divide a number of times before initiating their differentiation programme. The number of calretinin-expressing neurons increased further when cultures were treated with a combination of bFGF and retinoic acid. PMID- 9749799 TI - Presynaptic and postsynaptic localization of GABA(B) receptors in neurons of the rat retina. AB - The recently cloned GABA(B) receptors were localized in rat retina using specific antisera. Immunolabelling was detected in the inner and outer plexiform layers (IPL, OPL), and in a number of cells in the inner nuclear layer and the ganglion cell layer. Double-labelling experiments for GABA (gamma-aminobutyric acid) and GABA(B) receptors, respectively, demonstrated a co-localization in horizontal cells and amacrine cells. Electron microscopy showed that GABA(B) receptors of the OPL were localized presynaptically in horizontal cell processes invaginating into photoreceptor terminals. In the IPL, GABA(B) receptors were present presynaptically in amacrine cells, as well as postsynaptically in amacrine and ganglion cells. The postnatal development of GABA(B) receptors was also studied, and immunoreactivity was observed well before morphological and synaptic differentiation of retinal neurons. The present results suggest a presynaptic (autoreceptor) as well as postsynaptic role for GABA(B) receptors. In addition, the extrasynaptic localization of GABA(B) receptors could indicate a paracrine function of GABA in the retina. PMID- 9749800 TI - NT-4/5 and LIF, but not NT-3 and BDNF, promote NPY mRNA expression in cortical neurons in the absence of spontaneous bioelectrical activity. AB - Epigenetic factors are known to influence the differentiation of neocortical neurons. The present study analyses the role of spontaneous bioelectrical activity (SBA) and neurotrophic factors on the expression of neuropeptide Y (NPY) in rat visual cortical neurons using organotypic monocultures prepared from newborn animals and in situ hybridization to detect the NPY messenger ribonucleic acid (mRNA). Spontaneously active cortex cultures display NPY mRNA expression in about 7% of all cortical neurons from 10 days in vitro (DIV) on. Blocking the SBA by chronic application of 10 mM Mg2+ for 3-30 DIV reduces the percentage of NPY neurons to about 2%. Allowing an initial phase of SBA (1-20 DIV) followed by an SBA blockade (for 21-50 DIV) results in 2% labelled neurons, indicating a dramatic reduction of NPY mRNA expression in the absence of SBA. Surprisingly, the reverse experiment (a period of SBA blockade for 1-20 DIV followed by a period of SBA recovery for 21-40 DIV) does not cause an upregulation of NPY mRNA expression. However, allowing cultures to differentiate as spontaneously active cultures, then applying a transient period of SBA blockade which is followed by a second period of SBA, does rescue the NPY mRNA expression in 7% of the cortical neurons. We conclude that SBA is a main trigger for NPY mRNA expression and it is particularly important during an early postnatal period of differentiation. We then analysed whether neurotrophic factors known to modulate cortical neuropeptide expression are able to do so in the absence of SBA. Supplementing chronically blocked cultures with the neurotrophins, brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), neurotrophin-4/5 (NT-4/5) and the cytokine, leukaemia inhibitory factor (LIF), reveals that BDNF and NT-3 are unable to increase the percentage of NPY neurons. In contrast, LIF and NT-4/5 increase the percentage of NPY neurons to 4 and 6-7%, respectively. Moreover, neurons treated with NT-4/5 display a very high level of NPY mRNA expression in somata and in the dendritic trees. The data suggest a complex interplay and a hierarchy of epigenetic factors in regulating the neurochemical architecture of the developing neocortex. PMID- 9749801 TI - Region-specific expression of thyrotrophin-releasing hormone-degrading ectoenzyme in the rat central nervous system and pituitary gland. AB - Thyrotrophin-releasing hormone (TRH), a hypothalamic neuropeptide hormone and a putative neuromodulator/ neurotransmitter in the central nervous system is inactivated by the TRH-degrading ectoenzyme (TRH-DE), a TRH-specific metallopeptidase localized on the surface of neuronal brain cells in culture and on lactotrophic cells of the pituitary. After succeeding in cloning the cDNA of TRH-DE we now report on the cellular distribution pattern of this enzyme in rat brain, spinal cord and pituitary gland using in situ hybridization histochemistry. In the pituitary, TRH-DE mRNA was found both in the anterior and the neural lobe but not in the intermediate lobe. After treatment with triiodothyronine (T3) a dramatic increase in the mRNA levels of the TRH-DE and a decrease in the intensity of the TRH receptor could be observed in the anterior lobe of the pituitary. In brain, TRH-DE transcripts were predominantly found in neo- and allocortical regions with strongest signals in the olfactory bulb, the piriform cortex, the cerebral cortex, the granular layer of the cerebellar cortex and the pyramidal cells of the Ammon's horn. In the diencephalon, the highest TRH DE mRNA levels were observed in the medial habenulae followed by several hypothalamic subregions. In the mesencephalon and brainstem, moderate signals were present in the superior colliculi, substantia nigra, dorsal raphe and in the periolivar region. In the spinal cord, TRH-DE mRNA positive neurons were present in all layers. The very distinct distribution of TRH-DE in the brain and the hormonal regulation of the adenohypophyseal enzyme support the concept that this peptidase serves very specialized functions. PMID- 9749802 TI - Comparison of the electrophysiology and morphology of layers III and II neurons of the rat medial entorhinal cortex in vitro. AB - The basic membrane characteristics of neurons in layers II and III of the medial entorhinal cortex (MEA) were recorded using the intracellular current clamp technique in in vitro slices of the rat brain. Two types of cells were distinguished according to the presence of a time-dependent inward rectification (SAG current) with hyperpolarizing current pulses. The cells in which this inward rectification was not observed (No-SAG cells) had a larger input resistance, a more negative resting membrane potential and a more depolarized firing threshold. They more often displayed a strongly adapting firing pattern, and their action potentials had a slower decay rate and lacked a depolarizing afterpotential, compared with the SAG cells. SAG cells typically had a prominent rebound depolarization at the end of a hyperpolarizing current and membrane potential oscillations (7 Hz) upon subthreshold depolarizations. Cs+ blocked the time dependent inward rectification. The rebound depolarization persisted, even in the presence of tetrodotoxin. Biocytin labelling showed that layer III consisted mainly of pyramidal-shaped cells. Most layer III cells were of the No-SAG type. All cells in layer II, stellate and pyramidal cells, were classified as SAG cells. We conclude that the cells in MEA layers II and III display different electroresponsiveness, but that this appears to be more related to the layer where they are located than to a specific morphology. As layer III consisted mainly of cells of the No-SAG type, we suggest that layer III cells are less excitable than the SAG type layer II cells. PMID- 9749803 TI - Functional connectivity within the visual cortex of the rat shows state changes. AB - The aim of this study was to investigate the dynamics of the horizontal functional connectivity within the visual cortex during spontaneous activity or during visual stimulation. Two arrays of 16 electrodes were inserted in the visual cortex of a rat. From these electrodes a depth profile was obtained of the local spiking activity. The cross-correlations between all electrodes were estimated. Three types of cross-correlation peaks were identified and classified as; 'thin peaks', 'fast waves' and 'slow waves'. Partialization was applied, a mathematical method to reduce the amount of common input in correlations, and its effect on the three types of correlation peaks was studied. Slow waves were found to be the most vulnerable to partialization and thin peaks the least. From these observations it was concluded that the three types of peaks represent synchronous neuronal assemblies of different magnitude; slow waves large, fast waves intermediate and thin peaks assemblies composed of small numbers of neurons. Large changes were observed in the types of cross-correlations and their spatial distribution within the set of interarray combinations of electrodes. These changes were spontaneous, and could not be related to the visual stimulation. Two states were identified; the 'thin-peak' state and the 'slow-wave' state. The 'thin-peak' state is interpreted as occurring at a light level of anaesthesia and is characterized by the presence of thin peaks in all combinations of electrodes. Thin peaks with the largest strength were found in the upper interarray electrode combinations. The 'slow-wave' state is interpreted as occurring at a deep level of anaesthesia and is characterized by the presence of exclusively slow waves, which were limited mostly to the middle and lower interarray combinations of electrodes. Activation of the cortex is thus associated with the appearance of synchrony between small groups of neurons (thin peaks) which, in contrast to the slow-wave state, include the upper layers of the cortex. PMID- 9749804 TI - GFRalpha-3, a protein related to GFRalpha-1, is expressed in developing peripheral neurons and ensheathing cells. AB - We report here the identification of a gene, termed GFRalpha-3 (glial cell line derived neurotrophic factor family receptor alpha-3), related to GFRalpha-1 and GFRalpha-2 (also known as GDNFR-alpha and GDNFR-beta), and describe distribution of GDNFalpha-3 in the nervous system and other parts of the mouse body during development and in the adult. GFRalpha-3 in situ hybridization signals were found mainly in the peripheral nervous system, with prominent signals in developing dorsal root and trigeminal ganglia. Sympathetic ganglia were also positive. Developing nerves manifested strong GFRalpha-3 mRNA signals, presumably generated by the Schwann cells. Olfactory ensheathing cells were also positive. Other non neuronal cells appearing positive during development included chromaffin cells in the adrenal gland and small clusters of cells in the intestinal epithelium. In the central nervous system no robust signals could be detected at any stage investigated with the present probes. Compared with the previously described GFRalpha-1 and GFRalpha-2 mRNAs, which are widely distributed in the central nervous system and peripheral organs, the expression of GFRalpha-3 mRNA is much more restricted. The prominent expression in Schwann cells during development suggests a key role for GFRalpha-3 in the development of the peripheral nervous system. As Schwann cells are known to lack expression of the transducing RET receptor, we propose that a possible function of GFRalpha-3 during development could be to bind Schwann cell-derived GDNF-like ligands, thus presenting such molecules to growing axons. PMID- 9749805 TI - Reduced after-hyperpolarization in rat piriform cortex pyramidal neurons is associated with increased learning capability during operant conditioning. AB - Learning-related cellular modifications were studied in the rat piriform cortex. Water-deprived rats were divided to three groups: 'trained' rats were trained in a four-arm maze to discriminate positive cues in pairs of odours, 'control' rats were 'pseudo-trained' by random water rewarding, and 'naive' rats were water deprived only. In one experimental paradigm, the trained group was exposed to extensive training with rats learning to discriminate between 35 and 50 pairs of odours. Piriform cortex pyramidal neurons from 'trained', 'control' and 'naive' rats did not differ in their passive membrane properties and single spike characteristics. However, the after-hyperpolarizations (AHPs) that follow six spike trains were reduced after 'extensive training' by 43% and 36% compared with 'control' and 'naive', respectively. This effect was not observed in the piriform cortex of another group of rats, in which hyperexcitability was induced by chemical kindling. In another experimental paradigm rats were trained only until they demonstrated 'rule learning', usually after discriminating between one and two pairs of odours ('mild training'). In this experiment, a smaller, yet significant, reduction (20%) in AHPs was observed. AHP reduction was apparent in most of the sampled neurons. AHP remained reduced up to 3 days after the last training session. 5 days or more after the last training session, AHP amplitude recovered to pre-training value and did not differ between 'trained' rats and the others. Accordingly, training suspension for 5 days or more resulted in slower learning of novel odours. We suggest that increased neuronal excitability, manifested as reduced AHP, is related to the ability of the cortical network to enter a 'learning mode' which creates favourable conditions for enhanced learning capability. PMID- 9749806 TI - Efferent-mediated protection of the cochlear base from acoustic overexposure by low doses of lithium. AB - Many studies on anaesthetized animals and a few on awake animals have suggested that the cholinergic olivocochlear efferent feedback to outer hair cells can participate in the protection of the cochlea from acoustic overexposure. Lithium is known to stimulate acetylcholine synthesis and release in the brain and it is likely to act similarly at the level of the cochlear efferent synapses. We demonstrate here that, in the awake guinea-pig with a chronically implanted electrode on the round window of the cochlea, the temporary threshold shift induced by 1 minute exposure to different pure tones at around 90 dB sound pressure level (SPL) was reduced by as much as 40 dB, when exposure occurred after lithium treatment. The protection effect was not observed in anaesthetized animals. The effect was seen across the test frequency range of 6.4-12.5 kHz, suggesting that both 'fast' and 'slow' efferent effects are likely to be mediated by acetylcholine. Together our results provide new evidence that the olivocochlear efferents can provide a more efficient protection from acoustic overexposure when animals are awake. PMID- 9749808 TI - The new learner. PMID- 9749807 TI - Calcium influx through N- and P/Q-type channels activate apamin-sensitive calcium dependent potassium channels generating the late afterhyperpolarization in lamprey spinal neurons. AB - Lamprey spinal neurons exhibit a fast afterhyperpolarization and a late afterhyperpolarization (AHP) which is due to the activation of apamin-sensitive SK Ca2+-dependent K+ channels (KCa) activated by calcium influx through voltage dependent channels during the action potential (Hill et al. 1992, Neuroreport, 3, 943-945). In this study we have investigated which calcium channel subtypes are responsible for the activation of the KCa channels underlying the AHP. The effects of applying specific calcium channel blockers and agonists were analysed with regard to their effects on the AHP. Blockade of N-type calcium channels by omega-conotoxin GVIA resulted in a significant decrease in the amplitude of the AHP by 76.2+/-14.9% (mean +/- SD). Application of the P/Q-type calcium channel blocker omega-agatoxin IVA reduced the amplitude of the AHP by 20.3+/-10.4%. The amplitude of the AHP was unchanged during application of the L-type calcium channel antagonist nimodipine or the agonist (+/-)-BAY K 8644, as was the compound afterhyperpolarization after a train of 10 spikes at 100 Hz. The effects of calcium channel blockers were also tested on the spike frequency adaptation during a train of action potentials induced by a 100-200 ms depolarizing pulse. The N- and P/Q-type calcium channel antagonists decreased the spike frequency adaptation, whereas blockade of L-type channels had no effect. Thus in lamprey spinal cord motor- and interneurons, apamin-sensitive KCa channels underlying the AHP are activated primarily by calcium entering through N-type channels, and to a lesser extent through P/Q-type channels. PMID- 9749809 TI - Evaluation of an educational game for health sciences students. AB - An educational game was developed to promote student problem solving in the context of small group, problem-based learning (PBL). This study evaluated the game's effects on hypothesis and issue generation in PBL. Students in the Year III PBL course of a BScN program (N = 131) were stratified by program status and then randomly assigned to groups of 9 to 11 people per group. Half of the groups were randomly assigned to use the game (G); the remaining groups used the conventional method (CM) of generating issues and hypotheses. The G and CM groups crossed over for Term II. A Term I posttest demonstrated G groups had a higher proportion of accurate responses (85%) than CM groups (74%). A Term II posttest demonstrated no important differences between G and CM groups, suggesting the educational impact of the game persisted after students stopped using it. Strengths and limitations of the game as a learning aid also are addressed. PMID- 9749810 TI - Improving undergraduate nursing research education: the effectiveness of collecting and analyzing oral histories. AB - Nine oral histories of retired RNs (age range from 60 to 77) were collected by trained senior nursing students in their nursing research class. Attitudes toward nursing research of the 18 undergraduate baccalaureate nursing students participating in the collection and analysis of oral histories were compared with 20 students who did not participate. Both groups of students completed a pretest and posttest Attitudes Toward Nursing Research Questionnaire. The participating students had significantly positive changes in their attitudes toward nursing research. The nonparticipating group did not. While reminiscing about their careers, the retired RNs related historical events that influenced their nursing practice. Three themes emerged: World War II, technology, and intensive care units. Through an open-ended, written questionnaire following the interview, the nurses expressed positive benefits of participation. The collection and analysis of oral histories was an effective experiential learning strategy with positive outcomes for the students and the retired nurses who were interviewed. PMID- 9749811 TI - Clinical concept mapping as preparation for student nurses' clinical experiences. PMID- 9749812 TI - Path charting: a process and product for linking pathophysiological concepts. PMID- 9749813 TI - Cooperative education in nursing: a strategy for increasing marketability. PMID- 9749814 TI - Dear researcher: an alternative to the research critique. PMID- 9749815 TI - Crossword puzzles: adjunct clinical teaching strategy. PMID- 9749816 TI - Women's wellness: an innovative approach to nursing education. PMID- 9749817 TI - A health education initiative: teaching college women about osteoporosis. PMID- 9749818 TI - Classroom assessment: linking teaching and learning. AB - Formative evaluation can provide valuable data when evaluating student learning and the teaching strategies being used. Classroom assessment techniques as developed by Angelo and Cross are a type of formative assessment which involve ongoing assessment of student learning and assist faculty in selecting teaching techniques that are most effective in particular settings with specific groups of students. Classroom assessment techniques with applications to specific nursing education situations are included. PMID- 9749819 TI - Creating the teachable moment. PMID- 9749820 TI - Medication administration: does the teaching method really matter? AB - Do teaching methods affect students' performance of medication administration? This study was conducted to ascertain if there was a difference in baccalaureate nursing students' ability to accurately administer medication when taught using a faculty-assisted (control group) method versus a self-directed (experimental group) method. Student performance in medication administration was measured 1 week post laboratory practice using a 17-item instrument developed by the faculty. Students (N = 98) were randomly assigned to the faculty-assisted group or the self-directed group. The difference between the two groups was in the method of laboratory instruction. Students (n = 50) in the control group received faculty instruction on the skill performance during the laboratory practice. Students (n = 48) in the experimental group viewed a faculty-generated videotape on medication administration prior to laboratory practice. Performance evaluations were completed 1 week after the laboratory practice. Analysis of the data using a two-tailed t test for independent samples showed no significant difference (p < .05) between the two groups. PMID- 9749821 TI - Effect of rifampin on CD1b expression and double-negative T cell responses against mycobacteria-derived glycolipid antigen. AB - Non-classical antigen-presentation by CD1 molecules expressed on cytokine activated monocytes (CAM), and cell-mediated responses supported by double negative (DN) and by CD8+ responder alphabeta T cells, are involved in host resistance against mycobacterial infections. The CD1b protein is responsible for presentation of non-peptide, lipid antigens to T cells. In this context, a pivotal role is played by induction of CD1b protein on the membrane of human monocytes activated by GM-CSF alone, and more efficiently by GM-CSF combined with IL-4. Rifampin (RFP), a drug which is extensively utilized for chemoprophylaxis or treatment of Mycobacterium tuberculosis, is known to reduce a number of B, or T cell-dependent responses. Therefore we undertook immunopharmacological studies on RFP, to determine the effects of this agent on human macrophage function, relative to antigen presentation by CD1b molecules and on DN T cell cytolytic function. The results showed that: (a) graded concentration of RFP (2 or 10 microg/ml) induced a significant increase of CD1b expression, in CAM as evaluated by FACS analysis; (b) RFP increased significantly the specific mAb binding to CD1b on CAM surface; (c) treatment of effector cells with RFP did not reduce DN T cell-mediated cytolysis against lymphoblastoid cells transfected with CD1b cDNA (C1R.b6 cells), pulsed with M. tuberculosis. These results suggest that RFP could be of potential value in improving mycobacterial antigen presentation without impairing responder T cell function. PMID- 9749822 TI - Proliferative effects of oxidized low-density lipoprotein on vascular smooth muscle cells: role of dietary habits. AB - The effects were studied of native, partially-oxidized and totally-oxidized human low-density lipoprotein (LDL) on the proliferation of cultured rat aortic smooth muscle cells (VSMC), measured as an altered DNA synthesis. The LDL was obtained from three different human long-term diet groups (a control diet rich in saturated fats, a vegetarian diet, and a fish diet). The oxidized LDLs were prepared by oxidizing the LDL with copper sulfate. The DNA synthesis was measured by [3H]-thymidine incorporation into the DNA. The partially-oxidized LDL was the most potent promoter of DNA synthesis compared to the native or totally-oxidized LDL of the same diet group. The partially-oxidized LDL had a true mitogenic effect in the absence of exogenous growth factors. The native and totally oxidized LDL induced a significant increase in DNA synthesis, if they were obtained from the fish diet group. This study suggests an enhanced proliferative effect of partially-oxidized LDL on VSMC growth. PMID- 9749823 TI - Coronary pressure as a determinant of B-type natriuretic peptide gene expression in isolated perfused adult rat heart. AB - The role of coronary flow in the regulation of ventricular B-type natriuretic peptide (BNP) gene expression was studied in isolated perfused rat heart preparation. The increase of coronary flow from 5 ml/min to 20 ml/min for 2 h resulted in a 132+/-6 mm Hg increase in aortic perfusion pressure. The changes in BNP mRNA and immunoreactive BNP (IR-BNP) levels in response to hemodynamic stress were compared to those of c-fos and adrenomedullin (ADM) gene expression. The increase of coronary flow resulted in 1.5-fold increases in the left ventricular BNP mRNA (P < 0.001) and IR-BNP (P < 0.05) levels in 2-month old rats. There was also a 1.5-fold (P < 0.05) increase in ventricular c-fos mRNA levels, whereas ADM mRNA levels decreased by 74% (P < 0.001) in the left ventricle. In 18-month old rats, the increase in coronary flow decreased left and right ventricular BNP mRNA levels by 18% (P < 0.05) and 39% (P < 0.001), respectively. There were no changes in IR-BNP peptide and c-fos mRNA levels, whereas ADM mRNA levels decreased by 46% (P < 0.001) in the left ventricles. The results show that increased aortic perfusion pressure results in differential expression of cardiac genes including up-regulation of ventricular BNP and c-fos gene expression and down-regulation of ADM gene expression. Furthermore, aging seems to elevate the threshold at which hemodynamic stress of the heart results in a response at BNP gene level. PMID- 9749824 TI - Adrenomedullin suppresses atrial natriuretic factor (ANF) secretion from isolated atrium. AB - Atrial natriuretic factor (ANF) secretion was studied using isolated perfused right atria prepared from rats. Adrenomedullin (ADM), a recently identified 52 amino acid peptide whose biological activity has a striking resemblance to that of ANF, was added to the perfusate at a concentration of 1 microg/ml. The concentration of ANF secreted into the perfusate was measured by radioimmunoassay, under basal conditions (atrial wall unstressed), and during atrial distention (intraluminal pressure raised to 4 and to 6 cm water). It was found that basal secretion of ANF was not altered by ADM. However, when intraluminal pressure was raised, there was a significantly smaller increase in ANF secretion in the ADM-infused atria than in the control atria. It is concluded that ADM significantly reduces stretch-induced secretion of ANF, while having only minimal effects on basal secretion. Such an inhibitory mechanism would ensure the necessary negative feedback mechanism to counter the previously reported stimulatory actions of ANF on ADM secretion. Moreover, these results support the hypothesis that ADM could be responsible for the reduction in stretch induced ANF release observed during pregnancy. PMID- 9749825 TI - Effect of sulfhydryl oxidoreduction on permeability of cardiac tetrodotoxin insensitive sodium channel. AB - Effects of sulfhydryl oxidizing and reducing agents on permeability of the tetrodotoxin (TTX)-insensitive Na-channel were investigated in guinea-pig ventricular myocytes using the whole-cell patch-clamp technique. Mercury chloride (HgCl2) at 1-100 microM irreversibly blocked Na+ currents with no significant changes in the gating kinetics. In contrast, the hydrophilic sulfhydryl oxidizing agent, thimerosal at 50-100 microM little affected Na+ permeation through the Na channel. The Hg2+-induced block of Na+ current could be readily reversed by 1,4 dithiothreitol (DTT), an agent that reduces disulfide bonds. These results indicate that the formation of sulfur-Hg-sulfur bridge is essential for Hg2+ block. Pretreatment with DTT prevented the Hg2+ block of Na+ current, whereas Zn2+ and Cd2+ retained their abilities to block Na+ current after DTT treatment. An application of Zn2+ or Cd2+ resulted in the restoration of Hg2+ sensitivity of the DTT-treated channel. A conformational model for the Na-channel with multiple free sulfhydryl groups and native disulfide bonds could account for our experimental data regarding the effects of sulfhydryl modifying agents on the channel permeability. We conclude that the cardiac TTX-insensitive Na-channel contains functionally important free sulfhydryl groups and disulfide bonds which are accessible from the extracellular side by an aqueous pathway. These sulfhydryls would be capable of modulating the Na-channel permeability by affecting the conformation of channel pore region. PMID- 9749826 TI - Extracellular presence of IL-8 in the astrocyte-rich cultured cerebellar granule cells under acidosis. AB - In order to evaluate the functional role of chemotactic cytokines in the regulation of brain function, we examined the effects of acidosis on the production of IL-8 in cultured neurons and/or astrocyte-rich cerebellar granule cells as assessed by the ELISA method. A time-dependent and significant production of IL-8 was detected in the extracellular fluid of astrocyte-rich cultured cells at 2, 3 and 6 hrs after treatment with acidified Krebs-HEPES buffer (pH 6.9), although such production did not appear in the fluid of neuron rich cells. Additionally, microglia were detected by microscopic examination in both cultured cells under acidotic conditions. Only astrocyte-containing cultured cells produced a marked increase in intracellular IL-8 under acidotic conditions, although this production was much less than that seen in the extracellular fluid at 6 hrs under acidosis. The increase of IL-8 in astrocyte-rich cultures induced by acidosis was potentiated by treatment with glutamate, which enhanced the increase of cytosolic Ca2+ levels under acidosis, and was affected by extracellular Ca2+ conditions, by cyclosporine A, an inhibitor of calcineurin, and by trifluoperazine, an inhibitor of phospholipase A2. Significant inhibition of IL-8 production was detected after 6 hrs of pretreatment with trifluoperazine. Furthermore, the production of IL-8 under acidosis was associated with the appearance of astrocyte damage. These results suggest that Ca2+-dependent IL-8 is produced by astrocytes, but not neuronal cells, under acidosis, and that this production may be related to the process of cell dysfunction resulting from membrane destruction induced by acidosis. PMID- 9749827 TI - Nitroglycerin-induced aortic relaxation mediated by calcium-activated potassium channel is markedly diminished in hypertensive rats. AB - Nitroglycerin (NTG), a nitric oxide (NO) donor, is considered to relax vascular smooth muscle by stimulating soluble guanylate cyclase, which in turn increases cyclic GMP (cGMP) level. Recently it became evident that NO-induced vasodilatation is also mediated by stimulating Ca-activated K (K(Ca)) channels directly and/or indirectly through cGMP. We, therefore, tried to investigate the possible involvement or the alteration of K(Ca) channels in the mechanism of vasodilation induced by NTG in physiological and pathological conditions. Using rings prepared from thoracic aortas of spontaneously hypertensive rats (SHR) and those of age-matched Wistar-Kyoto rats (WKY), we studied changes in isometric tension of the rings in response to NTG to evaluate effects of a soluble guanylate cyclase inhibitor methylene blue (MB), and a specific blocker of K(Ca) channel charybdotoxin (CTX). Rings from WKY and SHR precontracted with norepinephrine showed similar aortic relaxation to NTG. MB markedly suppressed the NTG-induced relaxation in both strains, leaving about 30% of MB-resistant relaxation. CTX nearly completely eliminated this MB-resistant relaxation in WHY but did not affect this relaxation in SHR. These results suggest that NTG-induced vasorelaxation is mediated through i) cGMP-dependent and ii) cGM P-independent K(Ca) channel involving mechanisms, the latter may be diminished or virtually eliminated in hypertensive state. PMID- 9749828 TI - Localization of receptors for luteinizing hormone/chorionic gonadotropin in neural retina. AB - Although the expression of the luteinizing hormone (LH)/human chorionic gonadotropin (CG) receptor gene has been traditionally thought to be restricted to gonadal tissue, recent studies have shown that LH/CG receptors are present in many regions of the central nervous system (CNS), as well as in peripheral tissues. We now report the characterization of LH/CG receptor gene expression in the neural retina, a component of the CNS. Transcript levels in the retina are approximately equal to levels present in the cerebral cortex, but are at least 100 fold lower than the levels in testis. The density of LH/CG receptor transcripts, receptor protein and 125I-CG binding is the highest in the photoreceptor cells and then decreased throughout the inner retina. Our study is the first to demonstrate the presence of LH/CG receptors in the neural retina. This finding raises the possibility that photoreceptor cells have the potential to mount cellular responses to LH/CG that may impact on visual processing, and poses an intriguing connection to the proposed role of gonadotropins in the progression of proliferative retinopathy. PMID- 9749829 TI - Adenosine 5'-diphosphate as a factor in platelet aggregation induced by human plasma remnant lipoproteins. AB - The action of lipoprotein lipase on chylomicrons (CM) and very low density lipoproteins (VLDL) produces remnant lipoproteins (RLP) which are rich in triglycerides, cholesterol and apolipoprotein E (apo E). Apo E serves as a ligand for uptake of RLP by macrophages, platelets, endothelial cells and other cells expressing the LDL-receptor or the remnant receptor, thus having a major role in the clearance of plasma cholesterol and triglycerides, but at the same time, uptake of apo E-bearing RLP can profoundly alter the physiology of these cells and promote atherosclerosis. Like RLP, blood platelets also have roles in atherosclerosis and thrombosis, hence it is likely that RLP influence platelet activity as well. RLP derived from normal human plasma VLDL and CM were prepared using two monoclonal antibodies, anti-apo B-100 (JI-H) and anti-apo A-I (H-12) coupled to Sepharose 4B gel to form an immunoaffinity column. Lipoproteins containing apo B-100 including VLDL and LDL adsorb to (JI-H)-gel, while CM and HDL with apo A-I adsorb to (H-12)-gel. The particles in the unbound fraction (RLP) are rich in apo B-48, apo E and apo B-100 containing particles with multiple molecules of apo E. The RLP fraction with a total triglyceride of 14+/ 3.2 mg/ml; cholesterol, 0.39+/-0.1 mg/ml and protein, 0.78+/-0.24 mg/ml (n=19) was added to aliquots of blood of man, rabbits, guinea pigs, mice, and rats at protein equivalents of 0.98 to 78 microg/ml blood and agitated gently at 37 degrees C for 40 sec. Platelet aggregation was measured as a fall in single platelet count. RLP induced aggregation of platelets in man (p<0.005) rabbit (p<0.0005), guinea pig (p<0.002) and mouse (p<0.0001), but no RLP induced platelet aggregation was observed in the rat blood. Scanning electron microscopy revealed that in the presence of RLP, platelets had adhered to and formed aggregates on red cells. The platelet response to RLP was inhibited by apyrase known to scavenge ADP, by 5 microM 2-chloroadenosine, a platelet ADP receptor antagonist and by 3.4 microM cilostazol, a phosphodiesterase type III inhibitor known to raise cyclic AMP level in platelets. It is thought that RLP cause leakage of ADP from red cells which then mediates platelet aggregation. PMID- 9749830 TI - Tramadol induces antidepressant-type effects in mice. AB - Tramadol is a clinically-effective, centrally-acting analgesic. This drug is a racemic mixture of two enantiomers, each one displaying different mechanisms: (+)tramadol displays opioid agonist properties and inhibits serotonin reuptake while (-)tramadol inhibit preferentially noradrenaline reuptake. The action of tramadol on the monoaminergic reuptake is similar to that of antidepressant drugs. Therefore, we have examined the effects of (+/-)tramadol, (+)tramadol and (-)tramadol in a test predictive of antidepressant activity, the forced swimming test in mice. Both (+/-)tramadol and its (-) enantiomer displayed a dose dependent reduction on immobility; while the effect induced by the (+) enantiomer was not significant. Inhibition of noradrenaline synthesis, but not of serotonin synthesis, was capable of blocking the effect of (+/-)tramadol. The alpha adrenoceptor antagonist phentolamine, as well as the alpha2-adrenergic antagonist yohimbine, and the beta-adrenoceptor blocker propranolol countered the immobility reducing action of (+/-)tramadol. Moreover, neither the serotoninergic blocker methysergide nor the opioid antagonist naloxone antagonized the effect of (+/ )tramadol. Our results show that (+/-)tramadol and (-)tramadol have antidepressant-like effect in mice, probably mediated by the noradrenergic system rather than the serotoninergic or opioidergic ones. PMID- 9749831 TI - Is there a "non-MAO" macromolecular target for L-deprenyl?: Studies on MAOB mutant mice. AB - Several irreversible inhibitors of monoamine oxidase B, can promote survival of damaged neurons in several animal models of cerebral injury. Today it is evident that this effect is not a consequence of monoamine inhibition. Instead it has been proposed that L-deprenyl may act via an unrelated non-monoamine oxidase type of binding site. In the present study we have investigated if brain tissue from MAOB knockout mice contain such "non-MAO" binding sites for radiolabelled deprenyl. Interestingly, no binding of L-deprenyl was observed indicating that the mother compound does not act directly via a macromolecular target. This is enticing since several alternative mechanisms of action have been proposed in the literature. PMID- 9749832 TI - Direct repeat 3-type element lacking the ability to bind to the vitamin D receptor enhances the function of a vitamin D-responsive element. AB - In a previous study, we identified the element which allows the maximum response to 1,25(OH)2D3 in concert with two vitamin D-responsive elements (VDREs) in the rat 25-hydroxyvitamin D3 24-hydroxylase gene promoter, and designated it an accessory element [Ohyama, Y., Ozono, K., Uchida, M., Yoshimura, M., Shinki, T., Suda, T. and Yamamoto, O. Functional assessment of two vitamin D-responsive elements in the rat 25-hydroxyvitamin D3 24-hydroxylase gene. J. Biol. Chem., 1996, 271, 30381-30385]. The accessory element located adjacent to the proximal VDRE is not capable of binding to the vitamin D receptor (VDR), while its nucleotide sequence resembles the consensus sequence of VDREs, direct repeat 3 (DR3). To clarify the difference between the accessory element and VDREs, the function of the accessory element was compared with that of VDREs. The mutated accessory elements with a single nucleotide substitution showed the capability of binding to the VDR in vitro. However, these mutants still did not act as a VDRE when driven by the heterologous SV40 promoter. The accessory element did not enhance the function of a cAMP-responsive element. The corresponding site of the accessory element in the human 24-hydroxylase is a DR4-type element, and this element did not function as an accessory element. These results indicate that a critical nucleotide sequence is necessary for the binding to the VDR and for mediating the vitamin D effect, and suggest the different regulation between the rat and human 24-hydroxylase gene. PMID- 9749833 TI - Cloning, expression and characterization of rhesus macaque types 1 and 2 5alpha reductase: evidence for mechanism-based inhibition by finasteride. AB - The rhesus macaque types 1 and 2 5alpha-reductase (5aR1 and 5aR2) were cloned and expressed in COS cells to facilitate comparison of rhesus and human 5aRs. The deduced protein sequences of the rhesus SaRs shared 94% and 96% identity with the human type 1 and 2 isozymes, respectively. Despite a four amino acid insertion at the N-terminal region of rhesus 5aR1, the biochemical properties of rhesus and human homologs are very similar with respect to pH optimum, Km values for testosterone and progesterone, and inhibition by a variety of inhibitors. As expected, the biochemical properties of the human and rhesus 5aR2 are also very similar. The mechanism of inhibition of the rhesus 5aR1 and 5aR2 by finasteride was investigated in more detail. Finasteride displays time dependent inhibition of the rhesus 5aR1 and 5aR2 with second order rate constants of 4 x 10(3) M(-1) s(-1) and 5.2 x 10(5) M(-1)s(-1). Inhibition of rhesus 5aR2 with 3H-finasteride resulted in 3H bound to the enzyme which is not released by dialysis. Heat denaturation of the [rhesus SaR2:inhibitor] complex releases dihydrofinasteride, a breakdown product presumably related to the NADP+-adduct previously identified with the human SaRs (Bull et al., Mechanism-based inhibition of human steroid 5alpha-reductase by finasteride: Enzyme catalyzed formation of NADP dihydrofinasteride, a potent bisubstrate analog inhibitor. J. Amer. Chem. Soc., 1996, 118, 2359-2365). Taken together, these results provide good evidence that the rhesus macaque is a suitable model to evaluate the pharmacological properties of finasteride and other 5aR inhibitors. PMID- 9749834 TI - In vivo metabolism of the vitamin D analog, 22-oxacalcitriol: evidence for side chain truncation and 17-hydroxylation. AB - After intravenous administration of the vitamin D3 analog, 22-oxacalcitriol (OCT), to normal rats plasma metabolites were investigated by HPLC, GC-MS and LC MS. Five side-chain oxidation metabolites, 24R(OH)OCT, 24S(OH)OCT, (25R) 26(OH)OCT, (25S)-26(OH)OCT and 24oxoOCT, were identified by comparison with the corresponding synthetic compounds. These side-chain oxidation metabolites were similar to those of calcitriol [1alpha,25(OH)2 vitamin D3] described previously. Besides these five metabolites, two unique side-chain cleavage metabolites, 20S(OH)-hexanor-OCT and 17,20S(OH)2-hexanor-OCT, were identified as main metabolites in plasma by GC-MS and LC-MS using a specific chemical reaction. Our studies suggest that OCT is extensively metabolized and circulates in blood as a number of metabolites as well as unchanged OCT. This metabolism includes both unique pathways of C23-O22 cleavage and 17-hydroxylation, in addition to the side chain oxidation metabolites similar to those of 1alpha,25-(OH)2D3. PMID- 9749835 TI - Human estrogen sulfotransferase (hEST1) activities and its mRNA in various breast cancer cell lines. Effect of the progestin, promegestone (R-5020). AB - Using reverse transcriptase-polymerase chain reaction amplification it was possible to detect the presence of type 1 human estrogen sulfotransferase (hEST1) mRNA in the hormone-dependent: MCF-7 and T-47D, and hormone-independent: MDA-MB 231 and MDA-MB-468, human breast cancer cells. The expression of this mRNA is significantly higher in the MDA-MB-468 cells and a correlation of this mRNA expression with the enzymatic activity was observed. The progestin promegestone (R-5020) at a low concentration (5 x 10(-7) M) can significantly increase the estrogen sulfotransferase activity and its mRNA in the hormone-dependent MCF-7 and T-47D cells. As estrogen sulfates are biologically inactive, the stimulatory effect on sulfotransferase by promegestone may open attractive possibilities in the control of estradiol in human breast cancer. PMID- 9749836 TI - Using yeast to study glucocorticoid receptor phosphorylation. AB - The glucocorticoid receptor (GR) is a phosphoprotein and a member of the steroid/thyroid receptor superfamily of ligand dependent transcription factors. When the glucocorticoid receptor is expressed in yeast (Saccharomyces cerevisiae), it is competent for signal transduction and transcriptional regulation. We have studied the glucocorticoid receptor phosphorylation in yeast and demonstrated that the receptor is phosphorylated in both the absence and presence of hormone, on serine and threonine residues. This phosphorylation occurs within 15 min upon addition of radioactivity in both hormone treated and untreated cells. As reported for mammalian cells, additional phosphorylation occurs upon hormone binding and this phosphorylation is dependent on the type of the ligand. We have followed the hormone dependent receptor phosphorylation by electrophoretic mobility shift assay, and have shown that this mobility change is sensitive to phosphatase treatment. In addition, the appearance of hormone dependent phosphoisoforms of the receptor depends on the potency of the agonist used. Using this method we show that the residues contributing to the hormone dependent mobility shift are localized in one of the transcriptional activation domains, between amino acids 130-247. We altered the phosphorylation sites within this domain that correspond to the amino acids phosphorylated in mouse hormone treated cells. Using phosphopeptide maps we show that hormone changes the peptide pattern of metabolically labelled receptor, and we identify peptides which are phosphorylated in hormone dependent manner. Then we determine that phosphorylation of residues S224 and S232 is increased in the presence of hormone, whereas phosphorylation of residues T171 and S246 is constitutive. Finally, we show that in both yeast and mammalian cells the same residues on the glucocorticoid receptor are phosphorylated. Our results suggest that yeast cells would be a suitable system to study glucocorticoid receptor phosphorylation. The genetic manipulability of yeast cells, together with conservation of the phosphorylation of GR in yeast and mammalian cells and identification of hormone dependent phosphorylation, would facilitate the isolation of molecules involved in the glucocorticoid receptor phosphorylation pathway and further our understanding of this process. PMID- 9749837 TI - Hormonal regulation of the androgen receptor expression in human prostatic cells in culture. AB - The regulation of the androgen receptor (AR) expression was studied using immunocytochemical and Western blot techniques on separate cultures of epithelial cells (PNT2) and fibroblasts of human prostate. In both cell types, immunocytochemistry revealed both nuclear and cytoplasmic staining. Treatment with DHT (5 x 10(-9) M) increased both the intensity of nuclear staining and the number of cells stained. The increase, observed after DHT treatment was markedly decreased by cyproterone acetate (5 x 10(-7) M), confirming a direct action of DHT via the AR. This autoregulation of AR was confirmed by Western blot, and seems to involve transcription and protein synthesis, since it was suppressed by actinomycin D and cycloheximide. In fibroblasts, known to contain an estrogen receptor, estradiol treatment (5 x 10(-7) M) also increases the AR immunostaining. In addition, coculture studies show that epithelial cells require the presence of fibroblasts for optimal expression of the AR. These results demonstrate that prostate epithelial cells and fibroblasts have retained in culture, an hormonal sensitivity correlated with the presence of specific receptors and can serve as a model for the study of hormone action in this tissue in normal or pathological conditions. PMID- 9749838 TI - Site-directed mutagenesis identifies amino acid residues associated with the dehydrogenase and isomerase activities of human type I (placental) 3beta hydroxysteroid dehydrogenase/isomerase. AB - 3beta-hydroxysteroid dehydrogenase/steroid delta5-->4-isomerase (3beta HSD/isomerase) was expressed by baculovirus in Spodoptera fungiperda (Sf9) insect cells from cDNA sequences encoding human wild-type I (placental) and the human type I mutants - H261R, Y253F and Y253,254F. Western blots of SDS-polyacrylamide gels showed that the baculovirus-infected Sf9 cells expressed the immunoreactive wild-type, H261R, Y253F or Y253,254F protein that co-migrated with purified placental 3beta-HSD/isomerase (monomeric Mr=42,000 Da). The wild-type, H261R and Y253F enzymes were each purified as a single, homogeneous protein from a suspension of the Sf9 cells (5.01). In kinetic studies with purified enzyme, the H261R mutant enzyme had no 3beta-HSD activity, whereas the Km and Vmax values of the isomerase substrate were similar to the values obtained with the wild-type and native enzymes. The Vmax (88 nmol/min/mg) for the conversion of 5-androstene 3,17-dione to androstenedione by the Y253F isomerase activity was 7.0-fold less than the mean Vmax (620 nmol/min/mg) measured for the isomerase activity of the wild-type and native placental enzymes. In microsomal preparations, isomerase activity was completely abolished in the Y253,254F mutant enzyme, but Y253,254F had 45% of the 3beta-HSD activity of the wild-type enzyme. In contrast, the purified Y253F, wild-type and native enzymes had similar Vmax values for substrate oxidation by the 3beta-HSD activity. The 3beta-HSD activities of the Y253F, Y253,254F and wild-type enzymes reduced NAD+ with similar kinetic values. Although NADH activated the isomerase activities of the H261R and wild-type enzymes with similar kinetics, the activation of the isomerase activity of H261R by NAD+ was dramatically decreased. Based on these kinetic measurements, His261 appears to be a critical amino acid residue for the 3beta-HSD activity, and Tyr253 or Tyr254 participates in the isomerase activity of human type I (placental) enzyme. PMID- 9749839 TI - Effects of prolonged ACTH-stimulation on adrenocortical cholesterol reserve and apolipoprotein E concentration in young and aged Fischer 344 male rats. AB - Changes in the morphology of rat adrenal cortex with age include increased accumulations of lipid droplets and lipofuscin granules. Because glandular concentrations of cholesteryl esters (CE) and apolipoprotein (apo) E are also increased in parallel, the utilization or metabolism of lipid-droplet stored CE for steroidogenesis might be altered in aging cells. To explore this possibility, adrenocortical cholesterol storage and utilization were studied in 3-6 months-old (mo) (Y) rats and 20-23 mo (O) Fischer 344 male rats. Both groups received either adrenocorticotropin (ACTH1-39, Acthar gel) or gelatin alone daily for seven consecutive days. We found that: (a) the CE concentration in O rats, but not Y animals, was diminished by ACTH. The depleted CE in stimulated-O rats was replenished within five days post stimulation. Failure to deplete CE in stimulated-Y rats was not associated with an insufficient dose of the hormone, since stimulation of Y animals with higher doses of ACTH actually increased the CE concentration. In contrast, adrenocortical free cholesterol concentration remained constant during stimulation regardless of age. (b) The depleted CE in stimulated-O rats was principally comprised of cholesteryl adrenate, cholesteryl arachidonate and cholesteryl cervonate. The accumulated CE in stimulated-Y animals was primarily comprised of cholesteryl adrenate, cholesteryl arachidonate and cholesteryl oleate. (c) Whereas in stimulated-Y rats adrenal apoE concentration declined, the concentration in stimulated O animals was well maintained. (d) In vitro, adrenal homogenate or cytosolic fraction from stimulated-O rats displayed a higher capacity to hydrolyze exogenous CE than its Y counterpart. However, cholesterol esterification with external fatty acid substrates in adrenal homogenate or microsomal fraction was comparable in the two age-groups. Our findings revealed altered adrenocortical cholesterol reserve in O rats to cope with prolonged ACTH-stimulation. Changes in apoE levels and CE hydrolysis activity may be factors associated with this alteration. Depletion and accumulation of adrenocortical CE are reflected in parallel changes in cholesteryl adrenate and cholesteryl arachidonate, suggesting physiologic importance of these polyunsaturated fatty acids during sustained steroidogenesis. PMID- 9749840 TI - Adrenalectomy and dexamethasone treatment alter the patterns of basal and acute phase response-induced expression of acute phase protein genes in rat liver. AB - Hormonal requirements for full hepatic expression of alpha2-macroglobulin (alpha2M), alpha1-acid glycoprotein (AGP), haptoglobin (Hp) and gamma-fibrinogen (Fb) were assessed at the level of mRNA. Prior to exposure to turpentine-induced inflammation, rats were either depleted of glucocorticoids by adrenalectomy or supplemented with an excess of dexamethasone. Adrenalectomy alone did not affect the basal level of acute phase protein (APP) expression except for alpha2M mRNA, the level of which was enhanced. In contrast, dexamethasone treatment alone promoted full induction of alpha2M, significant, but not maximal increase of AGP and Hp mRNAs and suppression of Fb. In adrenalectomized rats, acute phase (AP) cytokines, released in response to inflammation, promoted full expression of Fb and Hp and increased the level of AGP mRNA whereas alpha2M mRNA remained at the basal level. Inflammation in dexamethasone pretreated rats elicited changes which, in comparison to mRNA values for dexamethasone unpretreated inflamed rats, were seen as overexpression of alpha2M, full expression of AGP and incomplete expression of Hp, whereas Fb mRNA remained at the basal level. These data suggest that glucocorticoids are the principal inducers of alpha2M and AP-cytokines of Fb. For full induction of AGP, additive actions of glucocorticoids and AP cytokines are required whereas expression of Hp is predominantly controlled by AP cytokines. PMID- 9749841 TI - Evolution of mammalian 11beta- and 17beta-hydroxysteroid dehydrogenases-type 2 and retinol dehydrogenases from ancestors in Caenorhabditis elegans and evidence for horizontal transfer of a eukaryote dehydrogenase to E. coli. AB - Physiological responses due to steroid hormones and retinoids are regulated by their cognate receptors and dehydrogenases. The origins of either regulatory mechanism are not fully understood. Here we examine the origins of the human 11beta-hydroxysteroid dehydrogenase-type 2, which regulates access of glucocorticoids to cells, and 17beta-hydroxysteroid dehydrogenase-type 2, which regulates access of androgens and estrogens to cells. Sequence comparisons trace their ancestry to homologs in Caenorhabditis elegans. These C. elegans proteins most closely resemble mammalian all-trans and 11-cis-retinol dehydrogenases. The similarity is sufficient -37% to 43% identity to suggest that one or more of the C. elegans homologs metabolizes a retinoid. Receptors for retinoids, but not for androgens, estrogens or glucocorticoids have been identified in C. elegans, suggesting that retinoid-mediated gene transcription is more ancient than that for adrenal and sex steroids. We propose that the hydroxysteroid dehydrogenase type 2 mechanism for regulating the androgen, estrogen and glucocorticoid concentrations in mammals descended from that for regulating retinoid concentrations. Interestingly, E. coli contains a protein with strong sequence similarity to mammalian retinol dehydrogenases. Sequence comparisons and phylogenetic analysis indicate that the E. coli protein may be an example of horizontal transfer from a eukaryote ancestor. PMID- 9749842 TI - Accurate evaluation of palpable breast masses by the triple test score. AB - BACKGROUND: We previously reported that the triple test (physical examination, mammography, and fine needle aspiration) for palpable breast masses yields 100% diagnostic accuracy when all 3 components are concordant (all benign or all malignant). However, 40% of cases are nonconcordant and require open biopsy. OBJECTIVE: To evaluate our experience with the triple test to develop a method to further limit the need for surgical biopsy. DESIGN: Diagnostic test study. SETTING: University hospital multidisciplinary breast clinic. PATIENTS: Two hundred fifty-nine patients with 261 palpable breast masses studied between 1991 and 1997. INTERVENTION: The triple test was prospectively applied to each breast mass. Each component of the triple test was assigned 1, 2, or 3 points for a benign, suspicious, or malignant result, respectively, yielding a total triple test score (TTS). MAIN OUTCOME MEASURES: The TTS was correlated with subsequent histopathologic examination results. RESULTS: Eighty-eight masses had a TTS of more than 6 points; all had malignant histopathologic characteristics. One hundred fifty-two masses had a TTS of 4 points or lower; all were benign. In both groups, diagnostic accuracy and predictive value were 100%, with P<.001. Twenty one masses had a TTS of 5 points; of these, 13 (62%) were benign and 8 (38%) were malignant. CONCLUSIONS: The TTS reliably guides evaluation and treatment of palpable breast masses. Masses that score 6 points or higher are malignant and should undergo definitive therapy; masses that score 4 points or lower are benign and may be clinically followed up. Only those masses that score 5 points (8% of our database) require open biopsy. PMID- 9749843 TI - Results of treatment of inferior vena cava syndrome with expandable metallic stents. AB - BACKGROUND: Patients with hepatic metastases often develop obstruction of the intrahepatic inferior vena cava (IVC), known as IVC syndrome. This obstruction is debilitating due to the development of ascites and anasarca. OBJECTIVES: To update our experience in the diagnosis and treatment of IVC syndrome and to evaluate the efficacy of expandable stents in the treatment of IVC syndrome. DESIGN: Retrospective review. SETTING: University hospital. PATIENTS: Twenty eight patients with hepatic metastases diagnosed as having IVC syndrome. INTERVENTION: Patients underwent transfemoral placement of Gianturco-Rosch self expandable Z metallic stents in the intrahepatic IVC. One patient was treated with a Wallstent. Stents were 15 to 25 mm in diameter and 60 to 140 mm in length. Pressure gradients across the IVC were measured before and after stent placement in all patients. MAIN OUTCOME MEASURES: Change in pressure gradient, relief of ascites and anasarca, loss of weight, patency of the primary stent, and survival after stent placement. RESULTS: Pressure gradients were reduced in all patients, which was followed by rapid reduction of ascites and anasarca with a median weight loss of 5.85 kg. Survival after stent placement varied from 1 to 99 days, with a mean of 34 days. Stent patency remained until death in all patients. CONCLUSION: The debilitation of IVC syndrome due to ascites and anasarca can be considerably palliated by placement of transfemoral percutaneous stents. PMID- 9749844 TI - Isolated transient loss of consciousness is an indicator of significant injury. AB - OBJECTIVE: To determine if isolated transient loss of consciousness is an indicator of significant injury. SETTING: University-based level I trauma center. DESIGN AND PATIENT: Phase 1 retrospective case series of all patients with trauma admitted directly from the emergency department to the operating room or an intensive care unit who had transient loss of consciousness as their only trauma triage criterion. Phase 2 prospective case series of all trauma patients transported by emergency medical system personnel with transient loss of consciousness as their only trauma triage criterion. MAIN OUTCOME MEASURES: Emergency operation and intensive care unit admission. RESULTS: Phase 1: From January 1, 1992, to March 31, 1995, we admitted 10255 patients with trauma. Three hundred seven (3%) met the enrollment criteria and were admitted to the operating room (n = 168) or intensive care unit (n = 139). Of these, 58 (18.9%) were taken to the operating room emergently to manage life-threatening injuries: 11 (4%) had craniotomies and 47 (15%) had non-neurosurgical operations. Phase 2: From July 1 to December 31, 1996, 2770 trauma patients were transported to our facility; 135 (4.9%) met the enrollment criteria. Forty-one (30.4%) of these required admission, and 6 (4.4%) were taken emergently to the operating room from the emergency department (1 [1%] for a craniotomy, 3 [2.2%] for intra-abdominal bleeding, and 2 [1.5%] for other procedures). Two (1.5%) of the 135 patients died. CONCLUSIONS: Patients with isolated transient loss of consciousness are at significant risk of critical surgical and neurosurgical injuries. These patients should be triaged to trauma centers or hospitals with adequate imaging, surgical, and neurosurgical resources. PMID- 9749845 TI - Is there a limit to massive blood transfusion after severe trauma? AB - OBJECTIVE: To examine the hypothesis that the futility of short-term care for trauma patients requiring emergency operation can be determined based on the number of units of blood transfused and associated risk factors. DESIGN: A 4-year retrospective review of a cohort of critically injured patients who underwent an emergency operation. SETTING: A large-volume, academic level I, urban trauma center. PATIENTS: One hundred forty-one consecutive patients received massive blood transfusions of 20 U or more of blood during preoperative and intraoperative resuscitation (highest, 68 U). There were 43 survivors (30.5%) and 98 nonsurvivors (69.5%). MAIN OUTCOME MEASURES: Mortality. RESULTS: The number of blood units transfused did not differ between survivors and nonsurvivors (mean +/ SD, 31 +/- 11 vs 32 +/- 10; P = .52). Stepwise multiple regression analysis identified 3 independent variables associated with mortality: need for aortic clamping, intraoperative use of inotropes, and intraoperative time with a systolic blood pressure of 90 mm Hg or less. However, blood usage was not different among the subgroups of patients who had 1 or more of these risk factors. When patients were stratified according to the amount of massive blood transfusion (20-29, 30-39, 40-49, and 50-68 U), the incidence of risk factors was not different across the 4 subgroups. Survival in the presence of risk factors was not affected by the amount of blood transfused. CONCLUSIONS: Although mortality among critically injured patients requiring operation and massive blood transfusion can be correlated with independent risk factors, discontinuation of short-term care cannot be justified based on the need for massive blood transfusion of up to 68 units. PMID- 9749846 TI - Thromboxane A2 in postischemic acute compartmental syndrome. AB - OBJECTIVE: To evaluate whether thromboxane A2 participates in the ischemia reperfusion injury associated with acute compartmental syndrome (ACS) and if by using a cyclooxygenase inhibitor this can be either reduced or abolished. DESIGN: To assess the role of thromboxane A2 in ACS, a tourniquet was applied for 2 hours to the hind limb of 12 dogs. Group 1 (n = 6) served as controls while group 2 (n = 6) was pretreated with lysine-acetyl-salicylate (Lysoprim). Blood thromboxane B2 levels and intracompartmental pressures were assayed prior to inflation of the tourniquet and at 5 minutes, 90 minutes, and 24, 72, and 144 hours after deflation. RESULTS: Five minutes after deflation, the compartmental pressure increased from 11.2 +/- 2.2 mm Hg to 16.1 +/- 3.3 mm Hg and 17 +/- 2.2 mm Hg (mean +/- SD) in groups 2 and 1, respectively. At 90 minutes and 24 hours, pressures were 17.1 +/- 3.3 mm Hg and 23.2 +/- 3.3 mm Hg (P<.01) and 15.3 +/- 2.6 mm Hg and 25.2 +/- 1.8 mm Hg (mean +/- SD) (P<.001), respectively, in groups 2 and 1. A similar effect, although of a lesser magnitude, was observed in the counterlateral limb. Thromboxane B2 levels increased from a mean (+/- SD) of 46 +/- 5.5 pg/0.1 mL to 132 +/- 7.5 pg/0.1 mL at 90 minutes in group 1, while remaining unchanged in group 2. CONCLUSIONS: Thromboxane A2 plays a major role in the ischemia-reperfusion injury of acute compartmental syndrome. By using a cyclooxygenase inhibitor both the levels of thromboxane and the compartmental pressures can be reduced. PMID- 9749847 TI - Manual vs robotically assisted laparoscopic surgery in the performance of basic manipulation and suturing tasks. AB - OBJECTIVE: To compare the surgical performance of manual and robotically assisted laparoscopic instruments on basic maneuvers and intracorporeal suturing in inanimate models. DESIGN: A set of laparoscopic tasks was used to evaluate basic endoscopic movements and intracorporeal suturing: positioning a cylinder on a Peg Board, dropping beads into receptacles, running a 25-cm rope, and capping a hypodermic needle. Intracorporeal knot tying and running a suture through predetermined points were evaluated separately. The sutures used for these tasks were 2-0 and 4-0 silk and 6-0 and 7-0 polypropylene. PARTICIPANTS: Twenty surgeons completed the set of laparoscopic tasks manually and then with a robotically assisted system. None had used the robotic system before. MAIN OUTCOME MEASURES: Time required to complete the tasks and the precision in performing them. RESULTS: The robotic system accurately reproduced the movements of the surgeons and filtered their hand tremors efficiently. In the basic tasks, operative times were significantly longer for the robotic system (P<.001). In the suturing tasks, operative times were longer with the use of the robotic system for sutures sizes 2-0 and 4-0 (P<.001). However, time differences were not significant for suture sizes 6-0 and 7-0 (P> or =.07). Precision measurements were similar for all tasks using the manual instruments and the robotically assisted system. No significant differences were found between the performance of advanced laparoscopic surgeons and laparoscopic fellows. CONCLUSIONS: Laparoscopic maneuvering and suturing is faster and just as precise when performed manually as when performed with the prototype robotic system. These differences in speed are inversely proportional to the size of the suture. Future generations of the robotic system may eliminate these differences. PMID- 9749848 TI - Surgeon-patient communication in the treatment of pancreatic cancer. AB - BACKGROUND: Effective physician-patient communication has been correlated with patient satisfaction and improved outcome. Pancreatic cancer (PC) is a disease with an overwhelmingly poor prognosis that requires a complex level of communication and emotional support. Since the treatment of PC is often surgical, surgeons play a central role in the care of these patients. OBJECTIVES: To assess the quality of long- and short-term surgeon-patient communication. To assess the role of the surgeon in the emotional support of patients with PC. DESIGN: Combined mail and telephone survey of a case series of patients who had undergone a pancreatic resection for PC. SETTING: Urban tertiary cancer referral center. PATIENTS: Forty-eight patients who underwent pancreatic resection for PC. INTERVENTION: Pancreatic resection. MAIN OUTCOME MEASURE: Patient satisfaction. RESULTS: Forty-eight patients completed surveys for a response rate of 70%. Patients were extremely satisfied with the information provided by their surgeon before surgery and while in the hospital. However, 21% of patients reported an unexpected outcome of their operation and 27% had questions about their disease at the time of the survey. Patients were largely satisfied with the emotional support they had received while in the hospital and after discharge. The attending surgeon was the most commonly desired source of additional emotional support. CONCLUSIONS: While surgeon-patient communication was extremely effective before surgery and during hospitalization, patients developed long-term questions and dissatisfaction after discharge from the hospital. Strategies to improve long term support and communication would benefit a significant number of patients with operable PC. Surgeons play an important role in the emotional support of patients with operable PC. PMID- 9749849 TI - Incidence of deep vein thrombosis after laparoscopic vs minilaparotomy cholecystectomy. AB - OBJECTIVES: To determine the frequency of deep vein thrombosis (DVT) associated with minimally invasive cholecystectomy and to determine, using minilaparotomy cholecystectomy as a control operation, the influence of the laparoscopic pneumoperitoneum on DVT formation. DESIGN: Prospective nonrandomized control trial. SETTING: Tertiary care university hospital. PATIENTS: One hundred consecutive patients intended to undergo either laparoscopic cholecystectomy (59 patients) or minilaparotomy cholecystectomy (41 patients) with either of 2 surgeons were prospectively enrolled between April 1996 and April 1997. The minilaparotomy cholecystectomy group served as controls to isolate the effect of the pneumoperitoneum. Patient details, operative details, and any thromboembolic or bleeding complications were recorded. The same thromboprophylaxis regimen was prescribed for each group; namely, preoperative and postoperative subcutaneous low-molecular-weight heparin (LMWH), graduated compression stockings, and intraoperative intermittent calf compression. INTERVENTION: Minimally invasive cholecystectomy. MAIN OUTCOME MEASURE: Frequency of DVT. Bilateral lower limb venous color duplex scanning was used to detect DVT. Scans were performed on 3 occasions: (1) preoperatively on admission to hospital, (2) on the first postoperative day, and (3) between 2 and 4 weeks postoperatively. RESULTS: Three patients in the laparoscopic group and 2 patients in the minilaparotomy group underwent conversion to conventional open cholecystectomy. There were no significant differences between patients in the 2 groups for age, sex, body mass index, preoperative white blood cell count, platelet count, prothrombin time, or activated partial thromboplastin time. There were no significant differences between the 2 groups for elective vs emergency operations, public hospital vs private hospital admissions, or consultant vs resident surgeon. Macroscopic gallbladder pathology grades for both groups were not significantly different, and there was no significant difference in the duration of postoperative hospital stay. Operative cholangiography was performed in a significantly larger proportion of laparoscopic cases (86% vs 66% in the minilaparotomy group; chi(2) test, P=.002), and the duration of anesthesia was significantly longer for the laparoscopic operation (118 minutes vs 98 minutes; t test, P=.05). Ninety-seven patients received preoperative LMWH and all patients received graduated compression stockings, intraoperative intermittent calf compression, and postoperative LMWH. Two of the 100 patients had postoperative DVT, 1 after laparoscopic cholecystectomy and 1 after minilaparotomy cholecystectomy. Both DVTs were detected by duplex examination on the first postoperative day. The DVT found after laparoscopic cholecystectomy was in 1 of the 3 patients who did not receive preoperative LMWH. There were no DVTs in any of the 40 patients who had an additional duplex scan between 2 and 4 weeks after operation. CONCLUSIONS: Despite the theoretical risk of thromboembolic disease due to use of the laparoscopic pneumoperitoneum, the frequency of DVT after either laparoscopic cholecystectomy or minilaparotomy cholecystectomy is low if adequate thromboprophylaxis is provided. PMID- 9749850 TI - Prospective randomized comparison of the Shouldice and Lichtenstein hernia repair procedures. AB - OBJECTIVE: To compare the Lichtenstein, tension-free mesh, and the Shouldice, 4 layer Bassini repair of the inguinal hernia. DESIGN: Prospective randomized clinical trial. SETTING: A private suburban hernia center. PATIENTS: Six hundred seventy-two men with inguinal hernias, aged 20 to 90 years, seen at the hernia center between January 1, 1990, and December 31, 1995. INTERVENTIONS: Slightly modified Shouldice and Lichtenstein repairs were used to repair primary and recurrent inguinal hernias. MAIN OUTCOME MEASURES: Recurrence rates, symptoms (including patient satisfaction), and infections. RESULTS: A total of 717 repairs in 672 patients, including 45 bilateral repairs, have been monitored to date. Recurrence of hernia occurred in 7 Shouldice repairs and 2 mesh repairs. Twelve superficial infections associated with Shouldice and 6 associated with mesh repairs were found. CONCLUSIONS: Both types of hernia repair are comparable and effective, but long-term results favor the Lichtenstein technique for reducing recurrences (to a P value of .10), ease of technical mastery, and application to the outpatient setting by use of a local anesthetic. PMID- 9749851 TI - Changing presentation and management of neutropenic enterocolitis. AB - OBJECTIVE: To characterize the current clinical presentation and management of neutropenic enterocolitis. DESIGN: Retrospective review of records of oncology unit patients requiring general surgical consultation for abdominal complaints in a 1-year period. SETTING: Oncology unit of a tertiary care, university teaching hospital. PATIENTS AND INTERVENTIONS: Fourteen patients diagnosed as having neutropenic enterocolitis were managed conservatively with operation reserved for failure of conservative therapy. MAIN OUTCOME MEASURES: Clinical data from patients at the time of presentation and during treatment for neutropenic enterocolitis. RESULTS: All 14 patients diagnosed as having neutropenic enterocolitis were receiving chemotherapy for solid tumors or leukemias. Seven patients were undergoing stem cell or autologous bone marrow transplantation. Presenting symptoms and physical examination findings were nonspecific. All patients except one had neutropenia at the time of diagnosis. Computed tomographic scans of the abdomen were the most useful confirmatory study for the diagnosis of neutropenic enterocolitis. All patients except one had resolution of neutropenic enterocolitis with conservative therapy. One patient whose course of conservative management failed had protracted neutropenia and required operation for resection of bowel with full-thickness necrosis. CONCLUSIONS: Neutropenic enterocolitis has evolved from a complication of patients with leukemia to a disease of patients receiving high-dose chemotherapy for many malignancies, solid as well as hematologic. Diagnosis of neutropenic enterocolitis continues to be a challenge, as patients typically present with nonspecific gastrointestinal tract symptoms. Neutropenia and computed tomographic scan findings are useful adjuncts in diagnosing neutropenic enterocolitis. Timely conservative treatment frequently allows resolution of neutropenic enterocolitis without operation. PMID- 9749852 TI - Determination of a safe vascular clamping method for liver surgery: evaluation by measuring activation of calpain mu. AB - OBJECTIVE: To determine the safest method of hepatic vascular clamping associated with the least ischemia-reperfusion injury of the liver during liver surgery. SETTING: University laboratories. SUBJECTS: Sixty-five adult male Wistar rats. METHODS: The hilar area of the left lateral and median lobes of rat liver was clamped for 10 minutes (group 1), 15 minutes (group 2), or 20 minutes (group 3) followed by 5 minutes of reperfusion. The procedure was repeated for a total period of ischemia of 60 minutes in each group. Control rats underwent laparotomy without vascular clamping. In addition to histological examination, we determined calpain mu activity, a marker of liver injury, by Western blotting using specific antibodies against the intermediate (activated) and proactivated forms of calpain mu. Measurements were performed at the end of ischemia and after 2 hours of reperfusion. We also determined the degradation of talin, an intracellular substrate of calpain mu, by Western blotting. RESULTS: The level of adenosine triphosphate and energy charge at 2 hours after reperfusion did not change after ischemia-reperfusion irrespective of the duration of ischemic cycle. After 60 minutes of intermittent ischemia followed by 2 hours of reperfusion, cell membrane bleb formation, calpain mu activation, and talin degradation were detected in groups 2 and 3 but not in group 1. CONCLUSION: The safest method of hepatic vascular clamping that produces a minimum or no ischemia-reperfusion injury is 60 minutes of 6 cycles of 10-minute vascular clamping interrupted by 5 minutes of reperfusion. PMID- 9749853 TI - Transfusion timing and postoperative septic complications after gastric cancer surgery: a retrospective study of 179 consecutive patients. AB - BACKGROUND: Immunosuppression associated with homologous blood transfusion was first observed in renal allograft transplantation. Clinical effects of transfusion-induced immunosuppression in surgical patients have been debated in the literature for more than a decade with contradictory results. OBJECTIVE: To investigate whether homologous blood transfusions significantly affect postoperative septic morbidity and mortality in patients undergoing elective surgery for gastric cancer. DESIGN: Case series. SETTING: Hospitalized care. PATIENTS: The hospital records of 209 patients who underwent elective surgery for gastric cancer at the Department of Surgery of the Hospital del Mar, Autonomous University of Barcelona in Spain, and at the Department of Surgery of the Catholic University of Rome in Italy from April 1984 to December 1990 were reviewed, and 179 patients were included in the study. MAIN OUTCOME MEASURES: The following variables were entered into univariate and multivariate analyses to identify factors potentially affecting postoperative septic morbidity: demographic data, weight loss, preoperative serum albumin level and lymphocyte count, type and duration of operative procedure, amount and timing of blood transfusion, and stage of disease. RESULTS: Univariate analysis showed that a large quantity of blood transfused (> 1500 mL) and transfusion in the postoperative period (group C) were associated with a worse clinical outcome. Postoperative transfusion was an independent predictor of septic morbidity in multivariate analysis. CONCLUSIONS: Despite transfusion-induced immunomodulation, homologous blood transfusion should not be considered a risk factor for postoperative septic morbidity in patients undergoing elective major abdominal surgery. The timing-response relationship between transfusions and septic morbidity in multivariate analysis may be the effect of uncontrolled confounders such as variation of volemia induced by stress response in patients who were developing or had just developed infectious complications. PMID- 9749854 TI - Feasibility of pylorus-preserving gastrectomy with a wider scope of lymphadenectomy. AB - OBJECTIVE: To demonstrate the feasibility and safety of pylorus-preserving gastrectomy (PPG) accompanied by complete suprapyloric and infrapyloric lymph node dissection. DESIGN: Retrospective review. SETTING: A university hospital in Japan. PATIENTS: Fifteen patients underwent PPG, and 28 patients underwent conventional distal gastrectomy (CDG) with Billroth I anastomosis. All patients had early gastric cancer, with either limited invasion in the mucosal layer or invasion into the submucosal layer. INTERVENTIONS: In the PPG procedure, the distal part of the stomach was resected while retaining a 1.5-cm pyloric cuff. The right gastroepiploic artery, the right gastric artery, and hepatic and pyloric branches of the vagus nerve were divided, and the infrapyloric artery was preserved. A modified D1 or D2 lymphadenectomy accompanied the PPG. MAIN OUTCOME MEASURES: Patients undergoing the PPG and CDG procedures were assessed 1 year after their surgical procedure. Changes in body weight, serum total protein levels, and serum albumin levels, the incidence of dumping syndromes, and endoscopic findings in the gastric remnant were compared between the 2 groups. RESULTS: Weight loss was significantly less in the PPG group than in the CDG group (P=.02). The incidences of early dumping syndromes, especially vasomotor symptoms, were significantly lower in the PPG group than in the CDG group (P=.03 and P=.02, respectively). The pyloric sphincter function was preserved, and there was no anastomotic leakage in the PPG group. CONCLUSIONS: The PPG procedure with complete D2 lymphadenectomy can be performed safely with a low incidence of major complications and a better postoperative outcome than the CDG procedure. The PPG procedure can be recommended for the treatment of early gastric cancer with broader indications. PMID- 9749855 TI - Partial or complete circular duodenectomy with highly selective vagotomy for severe obstructing duodenal ulcer disease: an initial experience. AB - OBJECTIVE: To evaluate partial and complete circular duodenectomy combined with highly selective vagotomy (HSV) for relief of gastric retention. DESIGN: A retrospective, case-comparison study. SETTING: University hospital referral center. PATIENTS: Eighteen patients with severe obstructing duodenal ulcer disease defined by failure of a saline load test and endoscopic narrowing of the gastric outlet to 5 mm or less. METHODS: In patients with severe obstructing ulcer the diseased duodenal segment was excised with electrocautery (partial excision, 10 patients; complete excision, 8 patients). An HSV was then done. Postoperative fasting gastric residuum measurement and measurement of the emptying of liquids and solids was done at 3 months and patients were weighed at 3 and 12 months. RESULTS: No patient experienced postoperative gastric retention or required reoperation in a 2-year follow up. The early emptying of liquid (20 minutes) in complete circular duodenectomy plus HSV was more rapid than in normal subjects and duodenal ulcer patients. The emptying of solids was slightly delayed in partial duodenectomy plus HSV compared with duodenal ulcer patients but not with normal controls. The emptying of solids in duodenal ulcer patients was more rapid than in normal controls. Weight gain was excellent at 3 and 12 months. CONCLUSION: Partial duodenectomy and complete circular duodenectomy plus HSV are more efficacious than alternative nonresective procedures in restoring gastric emptying to near normal and restoring weight in patients with obstructing duodenal ulcer. PMID- 9749856 TI - Augmented inflammatory responses and altered wound healing in cathepsin G deficient mice. AB - BACKGROUND: Cathepsin G is a neutral serine proteinase that exists primarily in azurophilic granules of neutrophils, but also as a proteolytically active membrane-bound form. While the specificity and many in vitro biological activities have been described for cathepsin G, little is known about the role of this enzyme in neutrophil function in vivo, particularly as it applies to the wound-healing process. OBJECTIVE: To determine the role of cathepsin G in cutaneous tissue repair by examination of full-thickness incisional wound healing in mice with a null mutation for cathepsin G. METHODS: Paired, full-thickness linear incisions were made on the backs of cathepsin G +/+ and cathepsin G -/- mice, and wound tissue was harvested at days 1, 2, 3, 5, 7, 10, and 14 after wounding. Neutrophil influx, myeloperoxidase activity, and migration were examined using light microscopy, the myeloperoxidase assay, and modified Boyden chamber technique, respectively. Wound-breaking strength was measured using tensiometry. RESULTS: The absence of cathepsin G led to a 42% decrease in wound breaking strength at day 7 after wounding (n=28; P<.002), which returned to the level of control mice by day 10 after wounding. Wound tissue sections in mice lacking cathepsin G also showed a 26% increase in neutrophil myeloperoxidase activity (n=12; P=.001) and an 18% increase in neutrophil influx (n=14; P=.002) at day 3 after wounding. Wound fluid collected on day 5 after wounding from cathepsin G-deficient mice attracted 58% more neutrophils than wound fluid collected from control mice (n=4; P<.05). CONCLUSIONS: Neutrophil cathepsin G is important during the early inflammatory stage of wound healing. Cathepsin G may be involved in processing 1 (or more) soluble mediator(s) in the wound milieu that is responsible for neutrophil chemotaxis. Our findings suggest that tight regulation of inflammation is necessary to prevent impaired healing during early tissue repair. PMID- 9749857 TI - Prediction and limitation of hepatic tumor resection without blood transfusion in cirrhotic patients. AB - BACKGROUND: The need for blood transfusion in cirrhotic liver resection is difficult to determine because of inaccurate estimation of operative blood loss. Moreover, blood transfusion is detrimental to cirrhotic patients. OBJECTIVE: To investigate the predictors and limitations of hepatectomy without blood transfusion for cirrhotic patients. DESIGN: Retrospective study. SETTING: University hospital, a tertiary referral center. PATIENTS: A consecutive 163 cirrhotic patients underwent resection for liver tumor(s) under a policy of restrictive blood transfusion. INTERVENTIONS: Estimated blood losses and clinicopathological features of patients who received and those who did not receive a blood transfusion were compared. MAIN OUTCOME MEASURES: Estimated operative blood losses, preoperative assessments, and operative procedures. RESULTS: There were 48 patients in the group who received a blood transfusion, with 1275 +/- 650 mL (mean +/- SE) of blood transfused, and 115 patients in the group who did not receive a blood transfusion. From discriminant analysis, the cutoff value of estimated blood loss for blood transfusion was 1685 mL. Tumor size and site of hepatectomy were found to be independent variables influencing blood transfusion under logistic regression analysis. CONCLUSIONS: Most cirrhotic patients tolerate hepatectomy without blood transfusion when the estimated operative blood loss is less than 1600 mL. Hepatectomy can be performed in cirrhotic patients without blood transfusion if the tumor is small (<5 cm), and/or the resection area is confined to Couinaud segments II, III, and VI. In this study, the largest amount of estimated blood loss in cirrhotic liver resection without blood transfusion was 2350 mL, but the uppermost limit remains to be determined. PMID- 9749858 TI - Laparoscopic cryoablation of hepatic metastases. AB - OBJECTIVE: To evaluate the feasibility of laparoscopic cryoablation for the management of hepatic metastases. DESIGN: Retrospective review. SETTING: Tertiary referral center. PATIENTS: Nine patients were evaluated by laparoscopy for planned laparoscopic cryoablation of hepatic metastases at The Cleveland Clinic Foundation, Cleveland, Ohio, from April 1996 to May 1997. RESULTS: Laparoscopic exploration revealed diffuse extrahepatic disease not identified by preoperative studies in 2 patients. The remaining 7 patients underwent 9 cryotherapy sessions. During 4 of the cryotherapy sessions, ultrasonography demonstrated unrecognized additional treatable hepatic lesions. An average of 3 lesions (range, 2-5) were treated. Operative time averaged 3.5 hours with a mean intraoperative blood loss of 235 mL. One patient had significant intraoperative hemorrhage requiring conversion to open hepatic resection for bleeding control. Eight of the 9 patients tolerated normal diets and ambulated independently on the first postoperative day. Following cryotherapy, 4 of the patients developed fever without an infectious source. One patient developed a postoperative bile leak requiring percutaneous biliary stenting. Postoperative hospital stay averaged 4.5 days (median, 4 days; range, 2-14 days). At a mean follow-up of 9 months, 4 of the 7 patients treated are alive without evidence of disease, 2 are alive with disease, and 1 patient with a pancreatic primary tumor has died of disease. CONCLUSIONS: Laparoscopy with laparoscopic ultrasonography is a useful tool in evaluating patients with hepatic metastases. Laparoscopic cryoablation is feasible and may result in lower postoperative morbidity in patients receiving aggressive treatment for inoperable hepatic metastases. PMID- 9749859 TI - Resection of pheochromocytoma under inferior vena caval isolation and extracorporeal charcoal hemoperfusion. AB - Resection of pheochromocytoma is associated with potential risks of hypertensive crises and serious arrhythmias due to massive release of catecholamines from the tumor. We report our surgical experience with complete inferior vena caval isolation and extracorporeal charcoal hemoperfusion (IVCI-CHP), which were performed to prevent systemic exposure to catecholamines during surgical manipulation. The IVCI-CHP significantly reduced the postfilter and systemic catecholamine levels compared with the prefilter levels (P<.01), indicating substantial catecholamine extraction (>90%) by the CHP filter. Reflecting the reduction of systemic exposure to catecholamines during IVCI-CHP, the patient's blood pressures were markedly stable. Our findings suggest that IVCI-CHP may be useful to minimize the risk of hypertensive crises during surgical manipulation of pheochromocytoma, by preventing systemic exposure to high levels of circulatory catecholamines. PMID- 9749860 TI - A randomized trial of an antibiotic- and antiseptic-coated central venous catheter in the prevention of catheter-related infections. PMID- 9749861 TI - Surgical reminiscences. I will never forget. PMID- 9749862 TI - Appendicitis: the quintessential American surgical disease. PMID- 9749863 TI - Lectin cytochemical analysis of lacrimal pleomorphic adenomas. AB - The activity of seven different types of biotinylated lectin were examined in normal and tumorous lacrimal gland tissue. In the normal lacrimal gland tissue, the glandular cells and the tubular epithelium were labeled by maclura pomifera agglutinin (MPA), soybean agglutinin (SBA), and bauhinia purpurea agglutinin (BPA). The myoepithelial cells were stained by griffonia simplicifolia agglutinin 1 (GS1). In the primary pleomorphic adenoma tissue, the epithelial components were labeled by MPA, ulex europaeus agglutinin, SBA, peanut agglutinin, and BPA. The mesenchymal components showed negative labeling. In contrast, the recurrent pleomorphic adenoma tissue showed a positive reaction in the mesenchymal components when GS1 and BPA were used. These results revealed differences in the glycoconjugate composition among normal and tumorous lacrimal gland tissues from patients with primary and recurrent pleomorphic adenomas. PMID- 9749864 TI - Possible prognostic markers in conjunctival dysplasia and squamous cell carcinoma. AB - Conjunctival squamous cell carcinoma (SCC) can develop from carcinoma in situ or severe dysplasia known as conjunctival intraepithelial neoplasia (CIN). Conjunctival intraepithelial neoplasia and SCC are histopathologically well defined conditions. However, it is difficult to determine the grading of dysplasia by clinical morphologic findings. Recently, proliferating cell nuclear antigen (PCNA) immunostaining, p53 immunostaining, and argyrophilic nucleolar organizer regions (AgNORs) staining have been established as valuable means of studying the biologic behavior of malignant cells. In the present study, these three staining techniques were used to examine histologic preparations of three conjunctival dysplasia and one SCC lesion. Five conjunctival tumor samples were obtained from four patients between July 1993 and October 1995. Following formalin fixation and embedding in paraffin, PCNA, and p53 immunostaining and AgNORs staining was performed with all tissue specimens. The PCNA-positive rate was the highest in SCC, followed by severe dysplasia and mild dysplasia. The p53 positive rate was the highest in severe dysplasia, followed by mild dysplasia, and negative in SCC. The AgNORs-count increased as malignancy advanced. These staining methods, which are markers for proliferative potency and cell differentiation, will be useful for early detection of changes in malignancy and will aid in decisions on treatment and prognosis. PMID- 9749865 TI - Cytokine production by T cells infiltrating in the eye of uveitis patients. AB - The capacity of T cells to produce cytokines was investigated using T-cell clones (TCCs) established from infiltrating cells in the aqueous humor (AH) or peripheral blood mononuclear cells (PBMC) of patients with Vogt-Koyanagi-Harada (VKH) disease or sarcoidosis. The cytokines produced and tested in the study were interleukin (IL)-1alpha, IL-6, IL-8, interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, and granulocyte monocyte colony stimulating factor (GM-CSF). All TCCs (n = 9) from AH of VKH patients spontaneously produced significantly larger amounts of IL-6, IL-8, and IFN-gamma than TCCs from healthy donor PBMC. All TCCs (n = 9) from AH of the sarcoidosis patient spontaneously produced significantly larger amounts of IL-1alpha, IL-6, and IL-8 than TCCs from healthy donor PBMC. In addition, the effects of antiinflammatory drugs on the cytokine production by the TCCs were investigated. Hydrocortisone significantly suppressed the production of IL-6, IL-8, and GM-CSF by TCCs from AH of VKH patients. Tacrolimus also significantly suppressed the production of IL-8 and GM-CSF by the TCCs. FTY720, an experimental drug, suppressed only GM-CSF production by TCCs from AH of VKH patients. Diclofenac failed to suppress the production of any cytokines by any TCCs. All tested drugs did not suppress the production of cytokines by TCCs from the sarcoidosis patient. These results thus suggest that cytokines produced by T cells infiltrating in the eye may play an important role in the pathogenesis of uveitis. PMID- 9749867 TI - Effect of intraocular irrigating solution on flicker electroretinogram during cataract surgery in human eye. AB - The effects of two commercially available intraocular irrigating solutions, Opeguard MA and BSS Plus, were studied during extracapsular cataract surgery in 45 eyes of 35 patients. After irrigation and aspiration of the residual cortex with Opeguard MA or BSS-Plus, the ERG amplitude increased, respectively, to 111.2+/-5.8% and 109.5+/-5.3% of the preirrigation amplitudes. The increases reached significance (116.9+/-7.0% and 115.7+/-6.5%; both P < .05) at the end of surgery compared with pre-irrigation ERG amplitudes. After irrigation with Opeguard MA or BSS-Plus, the ERG peak times were significantly prolonged to 103.9+/-0.8% and 104.2+/-1.2%, respectively, of the preirrigation peak times (both P < .01). The ERG peak times significantly shortened to 101.5+/-0.9% and 101.3+/-1.22%, respectively, at the end of surgery (P < .001 and P < .05) compared with just after irrigation. Although we have previously shown that Opeguard MA maintained amplitude and implicit time of 30 Hz flicker ERG during vitrectomy better than BSS-Plus, there were no statistically significant differences between the changes in amplitude and peak time with Opeguard MA and BSS-Plus during cataract surgery. We speculate that a drop in the retinal temperature during irrigation and aspiration in the anterior chamber and an increase in the photopic ERG amplitude during light adaptation with the operating microscope caused these ERG changes. PMID- 9749866 TI - Cyclic 3',5'-guanosine monophosphate synthesis induced by atrial natriuretic peptide, C-type natriuretic peptide, and nitric oxide in the rat retina. AB - This study was undertaken to determine whether pathways exist in the rat retina for atrial natriuretic peptide (ANP)-, C-type natriuretic peptide (CNP)-, and nitric oxide (NO)- cyclic 3', 5'-guanosine monophosphate (cGMP). Exposure of the retina to ANP (10(-7) mol/L), CNP (10(-7) mol/L), S-nitroso-N-acetylpenicillamine (10(-5) mol/L, SNAP; a NO donor), A23187 (10(-5)mol/L; a Ca2+ ionophore), and carbachol (10(-3) mol/L) caused 1.45 approximately 1.67-fold increases in cGMP content (P < .01). The increase in cGMP content induced by A23187 was blocked by 2-4-carboxyphenyl . 4455-tetramethyl imidazoline 1-oxyl 3-oxide (10(-3) mol/ L, carboxy-PTIO; a NO scavenger). Both carboxy-PTIO (10(-3) mol/L) and NG-nitro-L arginine (10(-3) mol/L, L-NNA: a NO synthase inhibitor) blocked the increase in cGMP content induced by carbachol. Atropine (10(-50 mol/L; a muscarinic receptor antagonist) also blocked the cGMP increase induced by carbachol. These data demonstrate that ANP-, CNP-, and NO-cGMP pathways exist in the rat retina and that the NO-cGMP pathway may be linked to the activation of the muscarinic receptor. PMID- 9749868 TI - Malignant melanomas of the iris. AB - The recognition of iris melanoma is important because a number of benign lesions clinically resemble these tumors. In this article, the epidemiological, clinical and histopathological features, treatment modalities, and prognosis of 41 iris melanoma patients, seen between 1964 and 1996 were evaluated. Of the patients, 20 were men and 21 women. Their mean age was 44.6 years. After determining the size, localization, and extension of the tumor, the management of choice was observation in 9, sector iridectomy in 15, iridocyclectomy in 6 and enucleation in 11 of the patients. During the follow-up, enucleation was also required in 6 and iridocyclectomy in 1 of the 7 patients who were in the observation or sector iridectomy group initially. Histopathologic examination revealed spindle cell in 27, mixed cell in 6 and epithelioid cell type melanomas in 2 of the 35 cases who underwent iridectomy, iridocyclectomy, and/or enucleation. The mean follow-up was 3.2 years and the mortality rate was found to be 2.4% during this period. One patient who died of metastases had epithelioid cell type melanoma. PMID- 9749869 TI - Effect of trabeculectomy on visual field in progressive normal-tension glaucoma. AB - The purpose of this study was to investigate the effectiveness of surgical treatment on the visual field in normal-tension glaucoma (NTG) eyes, and associated factors for visual field progression. Thirty-two patients (32 eyes) were enrolled in this retrospective study. Stepwise regression analysis was performed to correlate the visual field change with several clinical factors, such as intraocular pressure (IOP), and several IOP-unrelated parameters. Surgical reduction of IOP helps in preventing the progression of visual field damage in NTG eyes. The stepwise analysis revealed that better visual field prognosis was associated with higher preoperative IOP (P = 0.006) and the absence of diabetes mellitus (P = 0.04). The functional outcome in the lower IOP group was better when the patients were using Ca2+-channel blockers (P = 0.08); it was worse if disc hemorrhage appeared (P = 0.02). In conclusion, both IOP-related and unrelated factors are associated with visual field damage progression in NTG eyes. The functional prognosis in the eyes with low IOP depends on IOP-unrelated factors. PMID- 9749870 TI - Indocyanine green angiographic findings of lacquer cracks in pathologic myopia. AB - Lacquer cracks are thought to represent healed mechanical breaks in the retinal pigment epithelium, Bruch's membrane, and choriocapillaris complex. In this study, we analyzed the indocyanine green (ICG) angiographic features of lacquer cracks and compared them with findings using fluorescein angiography. Complete ophthalmologic examinations, fluorescein angiography, and ICG angiography were performed in 29 consecutive patients (37 eyes) with lacquer cracks. Fluorescein angiograms of the cracks revealed linear hyperfluorescence in all 37 eyes. Using ICG angiography, we observed linear hypofluorescence in all 37 eyes. In 15 of 37 eyes, the length of the hypofluorescent lesion detected by ICG angiography was longer than the hyperfluorescent lesion observed by fluorescein angiography. In 17 of 37 eyes, more lacquer cracks were observed by ICG angiography than by fluorescein angiography. These findings indicate that ICG angiography can detect the development of the lesion more precisely, and may provide useful information for diagnosing pathologic myopia. PMID- 9749872 TI - Group B streptococcal metastatic endophthalmitis in an elderly man without predisposing illness. AB - Our patient, an 83-year-old man, suddenly experienced acute lumbago and was prescribed bed rest. Later, pneumonia was diagnosed, even though he had no predisposing illness, and endophthalmitis developed in both eyes. Cultures of anterior chamber and vitreous specimens were positive for group B streptococcus. Treatment with systemic antibiotics, to which this bacteria is sensitive, was begun and his condition gradually improved. Nevertheless, the patient became blind in his right eye and the eye was enucleated. Histopathologic examination showed metastatic endophthalmitis with retinal detachment. Multiple microabscesses were found in the thickened choroid. We speculated that organisms disseminating from the microabscesses had caused the metastatic endophthalmitis. PMID- 9749871 TI - Indocyanine green angiography in classic choroidal neovascularization. AB - The indocyanine green (ICG) angiographic features of classic choroidal neovascularization (CNV) were evaluated in the 66 consecutive patients (70 eyes) by ICG angiography using the scanning laser ophthalmoscope. All patients had classic CNV documented by fluorescein angiography. Indocyanine green angiographic findings of classic CNV were as follows: Vessel architecture in 66% (46 of 70) of eyes, feeding vessels in 29% (20 of 70), and late hyperfluorescence in 93% (65 of 70) of eyes. Borders of classic CNV were found well-defined in 47% (33 of 70), and ill-defined in 49% (34 of 70) of eyes. In the remaining 4% (3 of 70) of eyes ICG angiography did not detect CNV. Our study indicates that fluorescein angiography remains the method of choice in the diagnosis of classic CNV. Indocyanine green angiography provides more information in the detection of feeding vessels of classic CNV. PMID- 9749873 TI - Risk of retinal detachment in patients with lattice degeneration. AB - To determine the risk of retinal detachment in patients with lattice degeneration of the retina, we statistically analyzed the incidence of retinal detachment in these patients. The data of hospital patients with retinal detachment associated with lattice degeneration in Kumamoto Prefecture, Japan, in 1990 were collected. The prevalence of lattice degeneration in Kumamoto was reported to be 9.5% in 1980. Based on population data from the 1990 census, the cumulative incidence of retinal detachment associated with lattice degeneration was calculated in this study. Among 1,840,000 residents in Kumamoto, there were 110 patients with retinal detachment associated with lattice degeneration; 72 with detachment resulting from tractional tears (tears), and 38 with detachment from atrophic holes. The cumulative incidence of retinal detachment from atrophic holes was 1.5% at the age of 40 years; from tears it was 3.6% at the age of 80 years. The cumulative incidence of detachment from both atrophic holes and tears was 5.3% at the age of 80 years. The results of this study are useful for clarifying the natural course of lattice degeneration. PMID- 9749874 TI - Cone electroretinograms in response to color stimuli after successful retinal detachment surgery. AB - Cone electroretinograms (ERGs) in response to different color flashes were examined using a Ganzfeld stimulus in 19 eyes after successful retinal detachment surgery. In the operated eyes, the short wavelength sensitive (S-) cone b-wave was reduced more than the mixed long (L-) and middle (M-) wavelength sensitive cone b-waves. The ratio of the S-cone ERG b-wave amplitude between operated eyes and fellow eyes was significantly lower than the L- and M-cone ERG b-waves (P < .01). These ERG results indicated that the S-cone system is more impaired than the L- and M-cone systems after retinal detachment surgery. PMID- 9749875 TI - Pathogenesis and neuroprotective treatment in Purtscher's retinopathy. AB - Purtscher's retinopathy is characterized by sudden visual loss in severely traumatized patients and is associated with multiple areas of superficial retinal whitening located primarily in the posterior pole. Visual outcome in Purtscher's retinopathy is variable, and there is no well-defined treatment. We report on a patient with immediate blurred vision in the right eye after a traffic accident. Ophthalmoscopy revealed multiple whitish patches scattered over the macular and peripapillary areas in the right eye. Fluorescein angiography showed multifocal retinal arteriolar occlusion in the early phase and staining of the involved retinal vessels and optic nerve head in the late phase. Indocyanine green angiography (ICG) showed rarefaction of choroidal vessels in the peripapillary area of the right eye at early phase. The late phase ICG study revealed multifocal hypofluorescent patches in the macular and peripapillary areas. Megadose steroid therapy was given with good visual response in the first 2 weeks, and the patient's vision had recovered completely when followed-up 10 months later. PMID- 9749876 TI - The management of retinal detachments associated with choroidal colobomas by vitrectomy with cyanoacrylate retinopexy. AB - The techniques used and the outcome in eyes treated for retinal detachment associated with choroidal coloboma are described. We reviewed the medical reports on five eyes of five patients with retinal detachment associated with choroidal coloboma who underwent vitrectomy. Retinal breaks were identified at the margin of or within the coloboma and the retina was successfully reattached by vitrectomy and cyanoacrylate retinopexy in four of the five eyes. The remaining one eye, with no visible retinal break both before and during surgery, also underwent cyanoacrylate retinopexy at regions suspected of retinal break, and was successfully reattached. In four eyes (80%) the vision showed improvement and had a visual acuity of 20/100 or better after surgery. None of the eyes required silicone oil tamponade or endophotocoagulation around the disc or at the papillomacular bundle. For the management of retinal detachment associated with choroidal coloboma, cyanoacrylate retinopexy is the method of choice, providing adequate chorioretinal adhesion and satisfactory visual outcome. PMID- 9749877 TI - Factors related to poor visual outcome in patients with retinopathy of prematurity after xenon are photocoagulation and/or cryocautery. PMID- 9749878 TI - Mechanisms of regulation of neurotensin receptors. AB - Since its discovery in 1973, the neuropeptide neurotensin has been demonstrated to be involved in the control of a broad variety of physiological activities in both the central nervous system and in the periphery. Pharmacological studies have shown that the biological effects elicited by neurotensin result from its specific binding to cell membrane neurotensin receptors that have been characterized in various tissue and in cell preparations. In addition, it is now well documented that most of these responses are subject to rapid desensitization. Such desensitization results in transient responses to sustained peptide applications, or to tachyphylaxis during successive stimulations in the same conditions. More recently, desensitization of neurotensin signalling was investigated at the cellular and molecular levels. In cultured cells, regulation at the second messenger level, receptor internalization, and receptor down regulation processes have been reported. These are proposed to play a critical role in the control of cell responsiveness to neurotensin. This review aims to compile recent data on the different biochemical processes involved in the regulation of the neurotensin receptor and to discuss the physiological consequences of this regulation in vivo. PMID- 9749879 TI - Pathophysiology of the kallikrein-kinin system in mammalian nervous tissue. AB - The nervous system and peripheral tissues in mammals contain a large number of biologically active peptides and proteases that function as neurotransmitters or neuromodulators in the nervous system, as hormones or cellular mediators in peripheral tissue, and play a role in human neurological diseases. The existence and possible functional relevance of bradykinin and kallidin (the peptides), kallikreins (the proteolytic enzymes), and kininases (the peptidases) in neurophysiology and neuropathological states are discussed in this review. Tissue kallikrein, the major cellular kinin-generating enzyme, has been localised in various areas of the mammalian brain. Functionally, it may assist also in the normal turnover of brain proteins and the processing of peptide-hormones, neurotransmitters, and some of the nerve growth factors that are essential for normal neuronal function and synaptic transmission. A specific class of kininases, peptidases responsible for the rapid degradation of kinins, is considered to be identical to enkephalinase A. Additionally, kinins are known to mediate inflammation, a cardinal feature of which is pain, and the clearest evidence for a primary neuronal role exists so far in the activation by kinins of peripherally located nociceptive receptors on C-fibre terminals that transmit and modulate pain perception. Kinins are also important in vascular homeostasis, the release of excitatory amino acid neurotransmitters, and the modulation of cerebral cellular immunity. The two kinin receptors, B2 and B1, that modulate the cellular actions of kinins have been demonstrated in animal neural tissue, neural cells in culture, and various areas of the human brain. Their localisation in glial tissue and neural centres, important in the regulation of cardiovascular homeostasis and nociception, suggests that the kinin system may play a functional role in the nervous system. PMID- 9749880 TI - The 90-kDa molecular chaperone family: structure, function, and clinical applications. A comprehensive review. AB - The 90-kDa molecular chaperone family (which comprises, among other proteins, the 90-kDa heat-shock protein, hsp90 and the 94-kDa glucose-regulated protein, grp94, major molecular chaperones of the cytosol and of the endoplasmic reticulum, respectively) has become an increasingly active subject of research in the past couple of years. These ubiquitous, well-conserved proteins account for 1-2% of all cellular proteins in most cells. However, their precise function is still far from being elucidated. Their involvement in the aetiology of several autoimmune diseases, in various infections, in recognition of malignant cells, and in antigen-presentation already demonstrates the essential role they likely will play in clinical practice of the next decade. The present review summarizes our current knowledge about the cellular functions, expression, and clinical implications of the 90-kDa molecular chaperone family and some approaches for future research. PMID- 9749881 TI - Prolactin: the forgotten hormone of human breast cancer. AB - Prolactin (PRL) is both a mitogen and a differentiating agent in the mammary gland. It has been shown to be involved in mammary cancer development in rodents, but in human breast cancer, its role has long been overlooked. Three criteria are applied to demonstrate PRL's involvement in this disease: (1) PRL receptors are present in human breast cancer cells, (2) human breast cancer cells in culture respond to PRL as a mitogen, and (3) PRL is synthesized by human breast cancer cells and inhibition of the binding of PRL to its receptors inhibits cell growth. PMID- 9749882 TI - Breech labor on the WHO partograph. AB - OBJECTIVES: To assess the impact of breech labor management using the WHO partograph on fetal and maternal outcomes of labor. METHOD: All 1,740 breech presentations in a larger multicenter hospital-based study in South East Asia of the use of the WHO partograph in labor management were studied. The partograph was introduced into each hospital during the study and a before and after analysis of various labor outcomes was conducted. RESULTS: There were 923 breech presentations prior to implementation of the partograph and 817 after. The overall Cesarean section rate was 29.7% (21.6% emergency and 7.6% elective). Introducing the partograph reduced Cesarean sections for multigravida from 27.1% to 19.3% (non-significant) but had no impact on the rate for primigravida (38.5% to 38.7%). Prolonged labor (> 18 hours) was reduced significantly among multigravida and primigravida (p < 0.05), despite a reduced use of oxytocin. Intrapartum stillbirths fell (non-significantly) from 1.9% to 1.1% (all parities combined). Fetal outcome, as measured by intrapartum deaths and Apgar scores < 7 at 1 minute, was significantly better (P < 0.05) when delivery was by Cesarean section rather than vaginally, regardless of use of the partograph. CONCLUSION: The use of the WHO partograph in the management of breech labor reduces prolonged labor and (among multigravida) Cesarean sections and improves fetal outcome. In this study, however, Cesarean section was a safer method of delivery for the baby, regardless of use of the partograph. PMID- 9749883 TI - Maternal sickle cell anemia and neonatal isoimmunization. AB - OBJECTIVE: To study the risk of alloimmunization in pregnant women with sickle cell disease (SCD) and of isoimmunization in their offspring. METHOD: Thirty mothers with SCD were studied and their 35 neonates (group 1) were compared with 538 infants of mothers without hemoglobinopathies (group 2). RESULT: Six mothers with SCD developed alloantibodies. There was no correlation with maternal age (P = 0.6), parity (P = 0.18) or blood transfusions (P = 0.4). The risk of alloimmunization by transfusion was 20%. Six neonates were isoimmunized, a higher incidence than in group 2 (P = 0.02; relative risk 3.07). Five had ABO incompatibility and only one had anti-c isoimmunization. All had reticulocytosis, jaundice and required phototherapy, one developed anemia but none required blood transfusion. CONCLUSION: Although alloimmunization was common in mothers with SCD and isoimmunization in their offspring, it was rarely due to non-ABO alloantibody. There was no significant neonatal morbidity. PMID- 9749884 TI - Fetal fibronectin as a predictor of preterm delivery in twin gestations. AB - OBJECTIVE: To evaluate fetal fibronectin as a predictor of premature delivery in twin pregnancies. METHOD: Cervicovaginal secretions were obtained from 52 pregnant women with twin pregnancies between 24 and 34 weeks of gestation. The secretions were analyzed to detect the presence of fetal fibronectin by immediate reading membrane test. The correlation between the presence of fetal fibronectin and preterm birth was evaluated. In addition, cervical dilatation and effacement were evaluated with each sampling. RESULT: The sensitivity, specificity, positive predictive value and negative predictive value to predict preterm delivery were 89.3, 50.0, 67.6, and 80.0%, respectively. A positive fetal fibronectin result was associated with a relative risk (RR) for preterm birth of 3.4 (95% CI, 1.2 9.5). A positive fetal fibronectin test associated with cervical dilatation or effacement increased the RR for preterm birth to 4.3 and 7.7, respectively, when compared with those with negative test and without cervical dilatation and effacement. CONCLUSION: Fetal fibronectin in the cervicovaginal secretions of patients with twin pregnancies is a sensitive predictor of preterm delivery. However, because of its low specificity, the fetal fibronectin test should be evaluated along with cervical changes for better identification of twins likely to develop preterm labor. PMID- 9749885 TI - Objective computerized fetal heart rate analysis. AB - OBJECTIVE: To assess the validity of a computerized methodology for cardiotocogram analysis based on a recently described reproducible visual estimation of the baseline. METHODS: Forty-two antepartum and 43 intrapartum cardiotocograms (CTGs) acquired by a personal computer were selected. Antepartum tracings were performed in the 48 h that preceded an elective cesarean section, and intrapartum tracings were performed until delivery. FHR baselines were estimated by an expert, according to an objective and reproducible methodology. Using these baselines, automated detection of accelerations and decelerations and estimation of variability was performed by the personal computer. A quantitative adaptation of the FIGO guidelines for fetal monitoring was used to classify tracings. Perinatal outcome was classified according to the Apgar score and umbilical arterial pH. Validity was then assessed by the proportions of agreement (PA), kappa statistic (K), sensitivity and specificity, with 95% confidence intervals (95% CI). Cases showing a disagreement between CTG and perinatal classification were reviewed and an adjustment in baseline definition was tested. RESULTS: The initial overall PA and kappa between CTG and perinatal classification were, respectively, 0.79 (95% CI: 0.69-0.87) and 0.62 (95% CI: 0.41-0.83). The overall PA and K, after baseline adjustment were, respectively, 0.89 (95% CI: 0.81-0.95) and 0.78 (95% CI: 0.58-0.98). Sensitivities and specificities ranged between 79% (95% CI: 60-92%) and 100% (95% CI: 95-100%). CONCLUSIONS: Good clinical prediction may be possible with an objective methodology for cardiotocogram analysis based on a recently described reproducible baseline estimation. PMID- 9749886 TI - Depression in women suffering perinatal loss. AB - OBJECTIVE: To analyze depression in women who have suffered perinatal loss in the present study. METHODS: The level of depression was studied, by means of the Beck Depression Inventory (BDI), in two groups of women, mothers who suffered perinatal loss and received psychological intervention for 1 year and mothers with live-birth babies. The BDI was recorded immediately after delivery and at 6 and 12 months postpartum. A third group of women with perinatal loss who received no intervention were studied only 12 months postpartum. RESULTS: At the time of delivery, women who suffered perinatal loss showed higher levels of depression, as measured by higher scores on the BDI than women experiencing a live-birth. At 6 months postpartum the intervention group showed improvement (lower BDI scores), but as a group they endorsed more depressive symptoms than the live-birth group. At 12 months the perinatal loss group who received the 1-year intervention was less depressed than the group who did not, and scored very similar to the live birth group. CONCLUSION: Women who experience perinatal loss endorse more depressive symptoms than mothers of live-births, and these depressive symptoms can be ameliorated by a psychological intervention. PMID- 9749887 TI - Sonomorphology of endometriotic cysts. AB - OBJECTIVE: The aim of the study was to evaluate prospectively the sonomorphological feature of histologically verified endometriotic cysts. METHODS: Transvaginal sonography was performed in 122 patients. Age distribution of the patients, and size and sonomorphology of the lesions were analyzed. RESULTS: Eighty-one percent of the endometriotic cysts occurred in patients between 31 and 50 years old with a peak of 29% between 31 and 35 years. Most of the cysts (81%) ranged between 30 and 59 mm in diameter. Forty-three percent of the endometriotic cysts were observed as monolocular cysts with internal echoes. More than half of the findings were observed to be multilocular, partially without any internal echos, partially even with solid parts or purely solid. CONCLUSION: The so-called 'typically' monolocular smooth-walled endometriotic cyst with internal echoes was only found in 43% of the cases. However, data from the literature show that one cannot assume even this special entity to represent endometriomas. Regarding all monolocular cysts with homogeneous internal echoes, one has to be aware of a great amount of functional cysts and a non-calculable residual risk of malignancy. PMID- 9749888 TI - Hysteroscopic endometrial ablation without endometrial preparation. AB - OBJECTIVE: To study the effectiveness of endometrial ablation by hysteroscopic resection without prior medical preparation for the treatment of women with persistent menorrhagia. METHOD: From January 1996 to January 1997, a total of 170 women with persistent menorrhagia and/or dysmenorrhea and who underwent hysteroscopic endometrial resection were included in the study. A thorough suction curettage was done before the procedure. The operation was conducted through a continuous flow hysteroscopic resectoscope with electrosurgery while the patient was under intravenous general anesthesia. The distention fluid used was 5% dextrose with a gravity feed infusion system consisting of a 2-1 bag between 1 and 1.5 m above the uterine cavity. After the procedure, the patients' conditions were followed for at least 6-18 months by telephone interview or at our clinic. RESULTS: A total of 127 women were available for a follow-up period of at least 6 months. Operative complications were 3%; three women had fever and received oral antibiotics; no uterine perforation occurred; one case of post operative bleeding was controlled by intrauterine balloon inflation; the average operation time was 21 min; The mean fluid deficit was 435 ml. Ninety-nine out of 127 women (78%) had adequately controlled menorrhagia (18.1% had amenorrhea, 42.5% hypomenorrhea and 17.3% had normal menstrual flow), while 27 women (21.2%) were failed due to unchanged or heavier menstrual flow after surgery. Eleven (40%) out of the 27 failed cases had myoma with menorrhagia, whereas only five women (5%) out of the 99 adequately treated women had myomas (P < 0.05). Thirty eight (54%) out of the 70 women with severe dysmenorrhea reported either lessening dysmenorrhea or no dysmenorrhea after the surgery. A total of 76 women (60%) were satisfied with the procedure. A second surgical procedure, either a resection or hysterectomy, was necessary in 13 women (10%) after ablation (seven received repeated ablations and six underwent hysterectomy). CONCLUSION: Endometrial ablation without endometrial suppression is a cheap, effective and acceptable procedure for treatment in women with persisted persistent menorrhagia. PMID- 9749889 TI - Pseudomyxoma peritonei--a heterogenous disease. AB - OBJECTIVE: To evaluate the origin of pseudomyxoma peritonei (PMP) in Chinese women. METHODS: The clinicopathologic features of 15 cases of PMP were reviewed. Immunostaining using antibodies for CK7 and CK20 was performed in the ovarian, appendiceal and peritoneal lesions of these cases. RESULTS: Appendiceal pathology was documented in five cases, including four mucinous cystadenoma and one simple mucocele. Eight ovarian tumors were found, including seven mucinous cystadenocarcinomas of low malignant potential and one mucinous cystadenoma. Synchronous ovarian and appendiceal lesions were discovered in three cases. One patient had adenocarcinoma of the pancreas. The origin of mucin production was not known in four cases with metastatic adenocarcinoma found in two of them. Immunoreactivity for CK20 was demonstrated in the tissues derived from the peritoneum, ovary, appendix and pancreas while only 23% (3 out of 13 women) of the peritoneal lesions and 33% (2 out of 6 women) of the ovarian tumors were immunoreactive for CK7. CONCLUSIONS: PMP is a heterogeneous lesion, which may develop from mucinous metaplasia of the peritoneum or from appendiceal, or ovarian lesions. Careful examination of the ovary and appendix with performance of appendectomy is advised in every case of PMP. Immunohistochemical examination of the peritoneal, ovarian or appendiceal lesions using antibodies, in particular that for CK7 would help in defining the origin of mucin production. PMID- 9749890 TI - Recurrent abortion and moderate or strong antiphospholipid antibody production. AB - OBJECTIVE: To investigate the treatment outcome for women suffering recurrent miscarriages associated with strong or moderate antiphospholipid antibody (aPL) production. METHODS: Sixty-seven pregnancies in 61 women demonstrating at least one kind of aPL with a history of recurrent miscarriages were treated with: (1) aspirin (ASA) alone; (2) prednisolone (PSL) and ASA; and (3) PSL, ASA, heparin and/or immunoglobulin (IgG). For comparison purposes the aPL-positive patients were divided into two groups, strongly and moderately-positive. IgG and IgM antibodies against PE and five negatively-charged phospholipids were measured by ELISA between 1987 and 1993. Beta2-glycoprotein I (beta2GPI) dependent anticardiolipin antibodies were measured by ELISA since 1993. Lupus anticoagulant was measured by a diluted aPTT method since 1993. RESULTS: Out of a total of 16 (50%) patients strongly-positive for aPL and 47 out of 51 (92.2%) moderately positive demonstrated a successful outcome. The live birth rate moderate group was significantly higher than in the strongly-positive cases (P < 0.0005). In the cases exhibiting moderate aPL production, 28 out of 30 (93.3%) receiving PSL and ASA and 14 out of 15 (93.3%) treated with ASA alone successfully gave birth. None of the 14 given ASA alone suffered preterm delivery or IUGR. In contrast 12 (36.4%) and 6 (18.2%) of the 33 patients treated with the PSL combination therapy suffered from preterm delivery and IUGR, respectively. CONCLUSIONS: The live birth rate in patients strongly positive for aPL is lower than that in patients with moderate aPL production even if treatment is performed during pregnancy. However, ASA is useful to treat cases with moderate aPL so that distinction of the two groups is warranted. PMID- 9749891 TI - Transperineal sonography in the first trimester of pregnancy. PMID- 9749892 TI - Beta-thalassemia: early antenatal diagnosis. PMID- 9749893 TI - ACOG educational bulletin. Osteoporosis. Number 246, April 1998 (replaces No. 167, May 1992). American College of Obstetricians and Gynecologists. AB - Advances in imaging technology allow clinicians to diagnose osteoporosis before a clinically significant fracture occurs. In addition, advances in pharmacology give clinicians a broad range of estrogens and nonestrogen treatments for osteoporosis. These advances in imaging technology and treatment are timely. As the population ages, a large increase in osteoporotic fractures is expected. It is key that women at increased risk are identified. Premenopausal and menopausal women should be encouraged to exercise and maintain adequate calcium intake. Menopausal women should be encouraged to consider estrogen replacement therapy, and those with risk factors who decline estrogen replacement therapy should be recommended for bone density measurements. Women with low bone density or established osteoporosis should be offered estrogen replacement, alendronate, or calcitonin therapy. Application of these new tools and treatments by obstetrician gynecologists may reduce the number of osteoporotic fractures experienced by our aging population. PMID- 9749894 TI - ACOG criteria set. Quality evaluation and improvement in practice. Cone biopsy of cervix. Number 32, April 1998. Committee on Quality Assessment. American College of Obstetricians and Gynecologists. PMID- 9749895 TI - Improving clinical pathway design: lessons learned from a computerised prototype. AB - Increasing costs of health care, fuelled by demand for high quality, cost efficient health care has propelled hospitals to restructure their patient care delivery systems. One such effort is the adaptation of an engineering project management methodology, the critical path method (CPM), as a tool to organise, standardise and improve the quality of healthcare delivery and hence patient outcomes. However, the two-dimensional nature and the size of paper impose severe limitations on the manual clinical pathways currently in use by hospitals. This paper analyses these inherent limitations and discusses some of the problems encountered in an attempt in early 1996 to create an electronic care map planner (CMP) based on the precedence diagramming method (PDM) model. It also reports on a current project to create a computerised clinical pathway tool to resolve the identified problems. PMID- 9749896 TI - Dynamic decision analysis in medicine: a data-driven approach. AB - Dynamic decision analysis concerns decision problems in which both time and uncertainty are explicitly considered. Two major challenges in dynamic decision analysis are on proper formulation of a model for the problem and effective elicitation of the numerous time-dependent conditional probabilities for the model. Based on a new, general dynamic decision modeling framework called DynaMoL (Dynamic decision Modeling Language), we propose a data-driven approach to addressing these issues. Our approach uses available problem data from large medical databases, guides the decision modeling at a proper level of abstraction and establishes a Bayesian learning method for automatic extraction of the probabilistic parameters. We demonstrate the theoretical implications and practical promises of this new approach to dynamic decision analysis in medicine through a comprehensive case study in the optimal follow-up of patients after curative colorectal cancer surgery. PMID- 9749897 TI - Combining laboratory data sets from multiple institutions using the logical observation identifier names and codes (LOINC). AB - A standard set of names and codes for laboratory test results is critical for any endeavor requiring automated data pooling, including multi-institutional research and cross-facility patient care. This need has led to the development of the logical observation identifier names and codes (LOINC) database and its test naming convention. This study is an expansion of a pilot study using LOINC to exchange laboratory data between Columbia University Medical Center in New York and Barnes Hospital at Washington University in St. Louis, where we described complexities and ambiguities that arose in the LOINC coding process (D.M. Baorto, J.J. Cimino, C.A. Parvin, M.G. Kahn, Proc. Am. Med. Inf. Assoc. 1997). For the present study, we required the same two medical centers to again extract raw laboratory data from their local information system for a defined patient population, translate tests into LOINC and provide aggregate data which could then be used to compare laboratory utilization. Here we examine a larger number of tests from each site which have been recoded using an updated version of the LOINC database. We conclude that the coding of local tests into LOINC can often be complex, especially the 'Kind of Property' field and apparently trivial differences in choices made by individual institutions can result in nonmatches in electronically pooled data. In the present study, 75% of failures to match the same tests between different institutions using LOINC codes were due to differences in local coding choices. LOINC has the potential to eliminate the need for detailed human inspection during the pooling of laboratory data from diverse sites and perhaps even a built-in capability to adjust matching stringency by selecting subsets of LOINC fields required to match. However, a quality standard coding procedure is required and examples highlighted in this paper may require special attention while mapping to LOINC. PMID- 9749898 TI - Providing dermatological photographs using the multimedia extension of the international computer database for radiation accident case histories. AB - The World Health Organisation (WHO) Radiation Emergency Medical Preparedness (REMPAN) centres have built up the International Computer Database for Radiation Accident Case Histories (ICDREC) to document the medical treatment of acute radiation syndrome (ARS) patients. Images play an increasing role as complementary information beside text and numerical data in medicine. In particular, retrieval and display of digitised skin photographs serve to improve patient care, medical education and scientific analysis concerning the cutaneous radiation syndrome (CRS). The ICDREC has been built up as a client/server system. Particular focus has been set on using commercial off-the-shelf software components. All the medical data including the multimedia data are stored in a relational database system. The database can be accessed by inexpensive personal computers in the physician's workplace. Retrieval of one photograph via local area network (LAN) requires approximately 3 s. Authorised institutions can access the database via the Internet. The current state of the ICDREC multimedia component is illustrated with the skin lesion treatment of a Chernobyl patient. An example is given on how the workflow in a dermatology department is supported by the ICDREC. PMID- 9749899 TI - Quantitative videodensitometric technique for verification of optimal coronary stent implantation. AB - Coronary angiography is not sensitive enough to define the results of stent implantation. Intravascular ultrasound defines accurately the anatomy of the vessel and the stent within the vessel and is thus considered the gold standard for defining the results of stent implantation. However intravascular ultrasound is an additional invasive procedure that is time consuming and expensive. This study describes a new simple quantitative videodensitometric technique, developed specifically to assess the results of stent implantation and compares the findings to intravascular ultrasound. In the proposed algorithm for the videodensitometric analysis, density profiles were constructed perpendicular to the long axis of the stented segment and each one was compared (after background subtraction) with a theoretic profile of a normal artery at that location. Density deficit index was determined at each point from the actual and theoretic profiles and a global volumetric density deficit index was calculated for each stent by integrating the deficit indices at all points along the stent. Similarly an area stenosis was determined at each point along the stent (using the stent and normal vessel cross sectional areas as defined by intravascular ultrasound) and the global volumetric stent stenosis was calculated by integrating the values of area stenosis along the stent. Twenty-five patients were evaluated immediately before and after coronary stent implantation. Global density deficit index improved from 66.1+/-16.4% before (after last balloon inflation) to 44.4+/-11.1% after stenting (P < 0.001). The shape of the curves of densitometric deficit indices along each stent were similar to the equivalent area stenosis curves as determined by intravascular ultrasound. The correlation (R = 0.74) between the global volumetric density deficit index and the global volumetric stent stenosis is statistically significant (P < 0.001). In conclusion, in this preliminary report we describe a new algorithm for videodensitometric analysis of the results of coronary stent implantation. As compared with intravascular ultrasound this method does not require an additional invasive procedure and it is quick cheap and easy to carry out. PMID- 9749900 TI - MERIT-9: a patient information exchange guideline using MML, HL7 and DICOM. AB - To realize clinical data exchange between healthcare providers, there must be many standards in many layers. Terms and codes should be standardized, syntax to wrap the data must be mutually parsable, then transfer protocol or exchange media should be agreed. Among many standards for the syntax, HL7 and DICOM are most successful. However, everything could not be handled by HL7 solely. DICOM is good for radiology images, but, other clinical images are already handled by other 'lighter' data formats like JPEG, TIFF. So, it is not realistic to use only one standard for every area of clinical information. For description of medical records, especially for narrative information, an standard generalized mark-up language, document type definition (SGML DTD) for medical information, called MML (medical markup language) had been created in Japan. It is already implemented in more than ten healthcare providers. However, it is again not realistic to use MML solely for clinical information in various level of detail. Therefore, we proposed a guideline for use of available medical standards to facilitate clinical information exchange between healthcare providers. It is called MERIT-9 (Medical Records, Images, Texts, -Information Exchange). A typical use is HL7 messages, DICOM files, referred from an MML file in a patient record, as external entities. Both MML and MERIT-9 have been research projects of Japanese Ministry of Health and Welfare and the purpose is to facilitate clinical data exchanges. They are becoming to be used in technical specifications for new hospital information systems in Japan. PMID- 9749901 TI - A microfabricated electrostatic haptic display for persons with visual impairments. AB - An electrostatic haptic display with three 7 x 7 electrode arrays of three different sizes was fabricated on a 4-in wafer using lithographic microfabrication techniques. The display utilizes electrostatic stimulation to generate a tactile sensation of texture on a scanning finger. The tactile sensation appeared to be a result of increased friction and vibration due to the electrostatic forces between the finger skin and the electrodes. Various spatial tactile patterns (lines, circles, squares, and triangles, etc.) can be presented on the display. Experiments of threshold, line separation, and pattern recognition were performed on subjects with visual impairments to study the spatial resolution and information transmission on arrays of variant electrode size and spacing. Two columns with two-column spacing can be resolved with 80% accuracy on the small array, for a spatial resolution of 5.8 mm in terms of edge to-edge electrode distance. The overall percentages of correct recognition for the patterns were 68.3, 72.1, and 71.3% on the small, medium, and large arrays, respectively. While subject is an important factor for both threshold and pattern recognition, electrode size was statistically significant for threshold only. Frequency and duty cycle of the stimulation waveform did not show statistical significance. PMID- 9749902 TI - Three-dimensional tactile display for the blind. AB - A tactile display device that can present tangible relief graphics for visually impaired persons has been developed. The tactile surface consists of a 64 x 64 arrangement of tactorpins with 3 mm interspacing. The tactor-pins are aligned in a hexagonal, rather than a square formation, to assure smooth depiction. The matrix has a total area of 200 mm x 170 mm. Each pin can be raised in 0.1 mm steps to a maximum height of 10 mm. Users can get certain information by touching the pins raised at varying heights with fingers and/or palms. Laboratory assessment of the device with six blind subjects showed its ability to transmit various kinds of information. PMID- 9749903 TI - Speech modifications algorithms used for training language learning-impaired children. AB - In this paper, the details of processing algorithms used in a training program with language learning-impaired children (LLI's) are described. The training program utilized computer games, speech/language training exercises, books-on tape and educational CD-ROM's. Speech tracks in these materials were processed using these algorithms. During a four week training period, recognition of both processed and normal speech in these children continually increased to near age appropriate levels. We conclude that this form of processed speech is subject to profound perceptual learning effects and exhibits widespread generalization to normal speech. This form of learning and generalization contributes to the rehabilitation of temporal processing deficits and language comprehension in this subject population. PMID- 9749904 TI - Impact of a poka-yoke device on job performance of individuals with cognitive impairments. AB - Job performance and production related issues are important not only to successful vocational training and ultimate job placement for individuals with cognitive disabilities, but also for their ability to have expanded vocational options. This study hypothesized that the application of Kaizen philosophy, and poka-yoke techniques in particular, could create job opportunities and improve productivity of individuals with cognitive disabilities. Poka-yoke or error proofing techniques are part of the collection of Kaizen techniques. Kaizen refers to continuous improvement in performance, cost/effectiveness, and quality. Kaizen strives to empower the worker, increase worker satisfaction, facilitate a sense of accomplishment, and thereby create pride-of-work. These techniques typically reduce the physical and cognitive demands of a task and thereby render the task more accessible. The job was a fuel clamp assembly. A redesigned assembly fixture was the poka-yoke intervention. Consistent with poka-yoke principles, the intervention improved the productivity of everyone attempting the assembly. In particular, the workers in this study showed an 80% increase in productivity and an average percent error drop from 52% to about 1% after the process redesign. Furthermore, the workers showed improved morale, self-esteem, and pride-of-work. Prior to the process redesign, only the higher functioning workers could successfully perform the assembly. After the redesign a greater number of workers could successfully perform the assembly. These results not only validated the study hypothesis, but demonstrated that the success facilitated by applying Kaizen techniques had similar results with individuals with cognitive disabilities as with nondisabled workers. PMID- 9749905 TI - Theoretical performance and clinical evaluation of transverse tripolar spinal cord stimulation. AB - A new type of spinal cord stimulation electrode, providing contact combinations with a transverse orientation, is presented. Electrodes were implanted in the cervical area (C4-C5) of two chronic pain patients and the stimulation results were subsequently simulated with a computer model consisting of a volume conductor model and active nerve fiber models. For various contact combinations a good match was obtained between the modeling results and the measurement data with respect to load resistance (less than 20% difference), perception thresholds (16% difference), asymmetry of paresthesia (significant correlation) and paresthesia distributions (weak correlation). The transversally oriented combinations provided the possibility to select either a preferential dorsal column stimulation, a preferential dorsal root stimulation or a mixed stimulation. The (a)symmetry of paresthesia could largely be affected in a predictable way by the selection of contact combinations as well. The transverse tripolar combination was shown to give a higher selectivity of paresthesia than monopolar and longitudinal dipolar combinations, at the cost of an increased current (more than twice). PMID- 9749907 TI - A photoelastic study of ligament strain. AB - The anterior cruciate ligament (ACL) in the human knee is known to exhibit as large as 10-20% of strain in association with knee flexion, while usual Hookean elastic materials exhibit at most 0.3%. Furthermore the complex ACL in shape deforms in highly variable manners due to the complicated three-dimensional (3-D) movements of the insertions in association with knee flexion. Such large and highly variable deformations of the ACL cannot be adequately quantified by one dimensional (1-D) and/or localized measurements. In order to measure strains over the entire area of the ACL, we employed the photoelastic coating method to analyze stress on the basis of the strains. A specific kind of polyurethane possessing optically high fringe-sensitivity was found to be most suitable for measurement purposes. From the preliminary experiments, it was found that a linear relation with 99% of correlation coefficient between the birefringence order and the strain on the polyurethane film continued up to 45% of strain. This study was done in two steps. In the first step, a study was done using a unique model of a knee joint fabricated in a way which allowed deformation and strain distribution in the model ACL to be directly observed and measured by means of the photoelastic coating method. It was confirmed that the film, having negligible effect on the mechanical properties of some biological soft tissues, closely reflected the strain behavior of the tissue. Thus in the second step, the photoelastic method was employed to measure the strains on an actual ACL during free flexion-extension of the knee. A specially designed apparatus was used to allow a natural motion of the knee. The photoelastic measurements demonstrated the potential utility of the application to measuring strain distributions not only on a model ligament but also on an actual ligament. The measurement of an actual ACL led the following results; The principal strain lines well depicted its fiber directions. A reciprocal function between the anterior and posterior bundles was observed. And there was a zero-strain area on the central and posterior sides near the tibial insertion. PMID- 9749906 TI - A mathematical model for skeletal muscle activated by N-let pulse trains. AB - A physiologically based mathematical model for skeletal muscle activated by neural impulses is presented. This model is developed specifically to capture the behavior for mammalian skeletal muscle activated by N-lets (sets of N high frequency pulses with variable interpulse intervals). N-let pulse trains have been demonstrated as a possible means of producing contractions with reduced fatigue and fiber-type transformation, while maximizing the force-time integral per pulse (FTIpP) of electrically stimulated muscle. This model is developed by modeling the underlying biophysical processes responsible for the initiation and maintenance of force generation in muscle. The release and reaccumulation dynamics of calcium ions from the sarcoplasmic reticulum are modeled and proposed as the governing mechanism for the observed N-let effects. It is found that the new model is robust, numerically stable and easily implemented. Simulation results are presented that demonstrate the model's ability to capture a variety of the nonlinear summation, force and stiffness variation effects seen experimentally when activating skeletal muscle with N-lets. General properties of FES muscle are also predicted by the model. The significant insight provided by this model into the internal dynamics of skeletal muscle is used to assess a variety of mechanisms proposed for N-let behavior. It is postulated that the calcium release and reaccumulation dynamics, as incorporated in this model, are responsible for the N-let effects found in experiment. PMID- 9749908 TI - A static dynamometer measuring simultaneous torques exerted at the upper limb. AB - The majority of available dynamometers are designed to measure force or torque in one specific direction, one joint at a time. For the quantification of motor incoordination in neurological patient populations, these dynamometers provide limited information about the global behavior of the limb under investigation. This report describes the potential use and function of a static dynamometer measuring torques exerted simultaneously at the shoulder (flexion-extension, abduction-adduction, internal-external rotation), elbow (flexion-extension), and forearm (pronation-supination). Orthogonal forces were measured at the arm and wrist using strain gauge transducers interfaced with a laboratory computer. The lever arms were specified to a software program and the joint torques were calculated in real time according to static equilibrium equations. The use of the dynamometer is illustrated by characterizing for one hemiparetic subject, the joints torques recorded at the shoulder, elbow, and forearm during isolated submaximal grip exertions at different force levels on both sides. The torques generated at the shoulder, elbow and forearm during the hand grip tasks on the affected side were significantly higher than those obtained on the nonaffected side and increased with the grip force level. These differences probably reflect the loss of movement selectivity observed following a lesion in the central nervous system. Further studies are currently being undertaken in neurological patient populations to characterize and quantify motor deficits using this dynamometer. As a long term goal, we hope that the method and technologies described here will contribute to the evaluation and rehabilitation of these populations. PMID- 9749909 TI - Separability of EEG signals recorded during right and left motor imagery using adaptive autoregressive parameters. AB - Electroencephalogram (EEG) recordings during right and left motor imagery can be used to move a cursor to a target on a computer screen. Such an EEG-based brain computer interface (BCI) can provide a new communication channel to replace an impaired motor function. It can be used by, e.g., patients with amyotrophic lateral sclerosis (ALS) to develop a simple binary response in order to reply to specific questions. Four subjects participated in a series of on-line sessions with an EEG-based cursor control. The EEG was recorded from electrodes overlying sensory-motor areas during left and right motor imagery. The EEG signals were analyzed in subject-specific frequency bands and classified on-line by a neural network. The network output was used as a feedback signal. The on-line error (100%-perfect classification) was between 10.0 and 38.1%. In addition, the single trial data were also analyzed off-line by using an adaptive autoregressive (AAR) model of order 6. With a linear discriminant analysis the estimated parameters for left and right motor imagery were separated. The error rate obtained varied between 5.8 and 32.8% and was, on average, better than the on-line results. By using the AAR-model for on-line classification an improvement in the error rate can be expected, however, with a classification delay around 1 s. PMID- 9749910 TI - EEG-based communication: improved accuracy by response verification. AB - Humans can learn to control the amplitude of electroencephalographic (EEG) activity in specific frequency bands over sensorimotor cortex and use it to move a cursor to a target on a computer screen. EEG-based communication could provide a new augmentative communication channel for individuals with motor disabilities. In the present system, each dimension of cursor movement is controlled by a linear equation. While the intercept in the equation is continually updated, it does not perfectly eliminate the impact of spontaneous variations in EEG amplitude. This imperfection reduces the accuracy of cursor movement. We evaluated a response verification (RV) procedure in which each outcome is determined by two opposite trials (e.g., one top-target trial and one bottom target trial). Success, or failure, on both is required for a definitive outcome. The RV procedure reduces errors due to imperfection in intercept selection. Accuracy for opposite-trial pairs exceeds that predicted from the accuracies of individual trials, and greatly exceeds that for same-trial pairs. The RV procedure should be particularly valuable when the first trial has >2 possible targets, because the second trial need only confirm or deny the outcome of the first, and it should be applicable to nonlinear as well as to linear algorithms. PMID- 9749911 TI - Design, control, and characterization of a Sliding Linear Investigative Platform for Analyzing Lower Limb Stability (SLIP-FALLS). AB - A novel device, the Sliding Linear Investigative Platform For Analyzing Lower Limb Stability (SLIP-FALLS), has been designed to study the detection and discrimination thresholds of humans to uniaxial horizontal step, ramp, or sinusoidal translations of the surface upon which they stand or stride. The device also can be used to test the human potential for, and mechanisms of, slips and falls. The SLIP utilizes air bearing technology and a noncontact linear motor to produce ultra-low-vibration translations. The FALLS system measures the forces on four load cells, platform linear and head tri-axial accelerations, four channels of electromyographic data, motor voltage, and a subject's psychophysical response; and derives other physiological and biomechanical measures, like center of-pressure and shear force. The effect of acceleration and shear force on the accuracy of the center-of-pressure calculations is presented. Operating ranges depend on the interactions among displacement, velocity, acceleration, and jerk parameters for linear translations, and between amplitudes and frequencies for sinusoidal translations. Displacements from 5 microm to 0.277 m, velocities from 5 microm/s to 0.3 m/s, and accelerations up to 2.5 m/s2 are achievable with precise control (i.e., without overshoot), but tradeoffs exist such that all three maxima cannot be reached simultaneously. For a 0.15 m/s linear translation at 4 m/s2, SLIP-FALLS produces substantially less vibration than the worm-driven NeuroTest system. The usefulness of having precise control over movement parameters is discussed. PMID- 9749912 TI - Effect of non-ionic detergents on apparent enzyme mechanism: V121A mutant of Streptomyces cholesterol oxidase endowed with enhanced sensitivity towards detergents. AB - One of the mutants of Streptomyces cholesterol oxidase with the Val121Ala mutation (V121A) was kinetically analysed. Although the reaction rate-substrate concentration curve of wild type follows a simple Michaelis-Menten equation, that of V121A is sigmoidal. The cooperativity was apparent and caused by non-ionic detergents that were used as a solvent of cholesterol. The concentration dependence of V121A on detergents was more significant than that of wild type, although the reaction rates of both enzymes decrease as the concentrations of detergents increase. Further experiments suggested that less hydrophobic interactions between V121A and detergents should be responsible for the apparent cooperativity. Since Val121 is in a hydrophobic loop located near the active site, the mutational effect is structurally discussed. PMID- 9749913 TI - Crystal structures of L201A mutant of D-amino acid aminotransferase at 2.0 A resolution: implication of the structural role of Leu201 in transamination. AB - The leucine-to-alanine mutation at residue 201 of D-amino acid aminotransferase provides a unique enzyme which gradually loses its activity while catalyzing the normal transamination; the co-enzyme form is converted from pyridoxal 5' phosphate to pyridoxamine 5'-phosphate upon the inactivation [Kishimoto,K., Yoshimura,T., Esaki,N., Sugio,S., Manning,J.M. and Soda,K. (1995) J. Biochem., 117, 691-696]. Crystal structures of both co-enzyme forms of the mutant enzyme have been determined at 2.0 A resolution: they are virtually identical, and are quite similar to that of the wild-type enzyme. Significant differences in both forms of the mutant are localized only on the bound co-enzyme, the side chains of Lys145 and Tyr31, and a water molecule sitting on the putative substrate binding site. Detailed comparisons of the structures of the mutant, together with that of the pyridoxamine-5'-phosphate form of the wild-type enzyme, imply that Leu201 would play a crucial role in the transamination reaction by keeping the pyridoxyl ring in the proper location without disturbing its oscillating motion, although the residue seems to not be especially important for the structural integrity of the enzyme. PMID- 9749914 TI - A re-estimation for the total numbers of protein folds and superfamilies. AB - The issue of the number of protein folds is steeped in controversy despite its significance for understanding evolution and predicting protein structure from amino acid sequence. Using various assumptions, several research groups have tackled this problem with very different results. In the present study, a more rigorous statistical approach is used to address this question. From three different data sets, the total number of protein folds is estimated to be about 650. A detailed theoretical analysis suggests that (i) a random sample of non transmembrane protein families has been selected for crystallization and structural determination, (ii) except for about 40 folds, most protein folds occurring in nature contain about the same number of different protein families. With the estimation of the total number of protein folds, the number of naturally occurring superfamilies can then be estimated as 1150. PMID- 9749915 TI - Misleading local sequence alignments: implications for comparative protein modelling. AB - Although it is well known that significant sequence similarity between proteins is reflected at the structural level, it is commonly assumed that any misaligned regions, as judged by the correct structure based alignment, are those where the local sequence identity is lower than the global. Recent studies have shown that this is not always the case and there can exist short stretches of high local identity which is not reflected in the structure based alignment. An analysis is presented of 290 pairs of homologous proteins with a view to quantifying the occurrence of these misleading local sequence alignments (MLSAs). It is found that such MLSAs are likely if the global sequence identity is less than 40% and can occur even when it is greater than 60%. The results have implications for automated homology modelling and also for the inference of function made by comparison. PMID- 9749916 TI - Main-chain conformational features at different conformations of the side-chains in proteins. AB - An analysis of the known protein structures has shown that the main-chain torsion angles, phi and psi of a residue can be affected by the side-chain torsion angle, chi1. The (chi1, psi) plot of all residues (except Gly, Ala and Pro) show six distinct regions where points are concentrated-although some of these regions are nearly absent in specific cases. The mean of these clusters can show a shift along the psi axis by as much as 30 degrees as chi1 is changed from around 180 to -60 to 60 degrees. Because of the lesser steric constraint points are more diffused along the psi axis when chi1 is approximately -60 degrees. Although points are more spread out along the phi axis in the (chi1, phi) plot, the dependence of phi on chi1 shows up in a shortened phi range (by about 30 degrees) when chi1 is around -60 degrees, and a distinct tendency of clustering of points into two regions when chi1 is approximately equal to 60 degrees, especially for the aromatic residues. Based on the dependence of the backbone conformation on its side-chain the 17 amino acids can be grouped into five classes: (i) aliphatic residues branched at the Cbeta position (although Thr is atypical), (ii) Leu (branched at the Cgamma position), (iii) aromatic residues (Trp can show some deviations), (iv) short polar residues (Asp and Asn), and (v) the remaining linear-chain residues, mainly polar. Ser and Thr have the highest inclination to occur with two different orientations of the side-chain that can be located through crystallography. Such residues exhibiting two chi1 angles have their phi and psi angles in a region that is common to the Ramachandran plots at the two different chi1 angles. The dependence of phi and psi angles on chi1 can be used to understand the helical propensities of some residues. Moreover, the average phi, psi values in the alpha-helices vary with the side-chain conformation. PMID- 9749917 TI - Folding motifs induced and stabilized by distinct cystine frameworks. AB - Bioactive peptides of different sources and biological functionalities, like endothelins, sarafotoxins, bee and scorpion venom toxins, contain a consensus cystine framework, Cys-(X)1-Cys/Cys-(X)3-Cys, which has been found to induce and stabilize a homologous folding motif named the cystine-stabilized alpha-helix (CSH). This is composed of an alpha-helical segment spanning the Cys-(X)3-Cys sequence portion that is crosslinked by two disulfide bridges to the sequence portion Cys-(X)1-Cys, itself folded in an extended beta-strand type structure. Search for sequence homologies of peptides and proteins in the SWISS-PROT and PDB data banks provided additional multiple examples of this type of cystine framework in serine proteinase inhibitors, in insect and plant defense proteins, as well as in members of the growth factor family with the cystine-knot. A comparative analysis of the known 3D-structures of these peptides and proteins confirmed that the presence of this peculiar cystine framework leads in all cases to a high degree of local structural homology that consists of the CSH motif, except for the cystine-knot, of the superfamily of the growth factors. In this case the cyclic structure formed by the parallel cysteine connectivities of Cys (X)1-Cys/Cys-(X)3-Cys framework is penetrated by a third disulfide bond with formation of a concatenated knot, and the two disulfide-bridged peptide chains Cys-(X)1-Cys and Cys-(X)3-Cys are located in beta-strands. Conversely, peptides and proteins containing Cys-(X)m-Cys/Cys-(X)n-Cys cystine frameworks that differ from m/n = 1/3 were found to fold only sporadically into local alpha-helical structures. PMID- 9749918 TI - Effect on thermostability and catalytic activity of introducing disulfide bonds into Aspergillus awamori glucoamylase. AB - Two additional disulfide bonds and three combined thermostabilizing mutations were introduced into Aspergillus awamori glucoamylase to test their effects on enzyme thermostability and catalytic properties. The single cysteine mutations N20C, A27C, T72C and A471C were made and combined to produce the double cysteine mutations N20C/ A27C and T72C/A471C. The double cysteine mutants were expressed efficiently in Saccharomyces cerevisiae, and disulfide bonds formed spontaneously after fermentation. At 50 degrees C, the single mutants N20C and A27C had decreased specific activity, whereas the specific activity of the double mutants N20C/A27C and T72C/A471C were similar to wild-type glucoamylase. The N20C/A27C mutation increased thermostability, with an increased activation free energy of 1.5 kJ/mol at 65 degrees C, while the single mutation A27C only slightly increased thermostability and N20C decreased it. The other disulfide bond-forming mutation T72C/A471C did not affect thermostability at pH 4.5. The N20C/A27C mutation was separately combined with two other thermostabilizing mutations, G137A and S436P. Thermostabilities of all of the combined mutated glucoamylases were additive. N20C/A27C/G137A glucoamylase had higher specific activity than wild-type glucoamylase from 45 to 67.5 degrees C. The disulfide bond between positions 20 and 27 connects the C-terminus of helix 1 and the following beta turn, suggesting that this region is important for glucoamylase thermostability. PMID- 9749919 TI - Converting trypsin to elastase: substitution of the S1 site and adjacent loops reconstitutes esterase specificity but not amidase activity. AB - The conversion of trypsin into a protease with chymotrypsin-like activity and specificity required substitution of fifteen residues in the S1 site and two surface loops with their chymotrypsin counterparts [Hedstrom,L., Szilagyi,L. and Rutter,W.J. (1992) Science, 255, 1249-1253]. These residues may define a set of general structural determinants of specificity in the trypsin family. In order to test this hypothesis, we have attempted to convert trypsin into a protease with specificity for substrates containing small aliphatic residues by replacing the S1 site and these surface loops with the analogous residues of elastase. Five elastase-like mutant enzymes were constructed with various combinations of these substitutions. Four mutant enzymes catalyze the hydrolysis of MeOSuc-Ala-Ala-Pro Ala-SBzl more efficiently than the hydrolysis of Suc-Ala-Ala-Pro-Phe-SBzl. This observation indicates that the mutant enzymes have elastase-like esterase specificity. The best mutant, Tr-->E1-2, is a more specific esterase than elastase: the ratio of the values of kcat/Km for MeOSuc-Ala-Ala-Pro-Ala-SBzl and Suc-Ala-Ala-Pro-Phe-SBzl is greater than 160 for Tr-->E1-2 and 50 for elastase. However, the esterase activity of Tr-->E1-2 is 300-fold less than elastase; in addition, Tr-->E1-2 has no measurable amidase activity. Thus these substitutions do not construct a protease with elastase-like activity. These experiments indicate that a unique structural solution is required for each different specificity. Previous work suggested that instability of the S1 site is a major barrier to redesigning the specificity of trypsin. This view is corroborated by preliminary structural studies of Tr-->E1-2. One dimensional 1H NMR spectrum of Tr-->E1-2 suggests that the S1 site and the two surface loops of this mutant trypsin may be disordered. PMID- 9749920 TI - Rational design of Rhizopus oryzae lipase with modified stereoselectivity toward triradylglycerols. AB - The binding site of sn-1(3)-regioselective Rhizopus oryzae lipase (ROL) has been engineered to change the stereoselectivity of hydrolysis of triacylglycerol substrates and analogs. Two types of prochiral triradylglycerols were considered: 'flexible' substrates with ether, benzylether or ester groups, and 'rigid' substrates with amide or phenyl groups, respectively, in the sn-2 position. The molecular basis of sn-1(3) stereoselectivity of ROL was investigated by modeling the interactions between substrates and ROL, and the model was confirmed by experimental determination of the stereoselectivity of wild-type and mutated ROL. For the substrates, the following rules were derived: (i) stereopreference of ROL toward triradylglycerols depends on the substrate structure. Substrates with 'flexible' sn-2 substituents are preferably hydrolyzed at sn-1, 'rigid' substrates at sn-3. (ii) Stereopreference of ROL toward triradylglycerols can be predicted by analyzing the geometry of the substrate docked to ROL: if the torsion angle phiO3-C3 of glycerol is more than 150 degrees, the substrate will preferably be hydrolyzed in sn-1, otherwise in sn-3. For ROL, the following rules were derived: (i) residue 258 affects stereoselectivity by steric interactions with the sn-2 substituent rather than polar interactions. To a lower extent, stereoselectivity is influenced by mutations further apart (L254) from residue 258. (ii) With 'rigid' substrates, increasing the size of the binding site (mutations L258A and L258S) shifts stereoselectivity of hydrolysis toward sn-1, decreasing its size (L258F and L258F/L254F) toward sn-3. PMID- 9749921 TI - Calorimetric study of mutant human lysozymes with partially introduced Ca2+ binding sites and its efficient refolding system from inclusion bodies. AB - During the process of evolution, ancestral lysozymes evolved into calcium-binding lysozymes by acquiring three critical aspartate residues at positions 86, 91 and 92. To investigate the process of the acquisition of calcium-binding ability, two of the aspartates were partially introduced into human lysozyme at positions 86, 91 and 92. These mutants (HLQ86D, HLA92D and HLQ86D/D91Q/A92D), having two critical aspartates in calcium-binding sites, were expressed in Escherichia coli as non-active inclusion bodies. For the preparation of lysozyme samples, a refolding system using thioredoxin was established. This system allowed for effective refolding of wild-type and mutant lysozymes, and 100% of activity was recovered within 4 days. The calcium ion dependence of the melting temperature (Tm) of wild-type and mutant lysozymes was investigated by differential scanning calorimetry at pH 4.5. The Tm values of wild-type, HLQ86D and HLA92D mutants were not dependent on calcium ion concentration. However, the Tm of HLQ86D/D91Q/A92D was 4 degrees higher in the presence of 50 mM CaCl2 than in its absence, and the calcium-binding constant of this mutant was estimated to be 2.25(+/-0.25)x10(2) M(-1) at pH 4.5. Moreover, the calcium-binding ability of this mutant was confirmed by the result using Sephadex G-25 gel chromatography. These results indicate that it is indispensable to have at least two aspartates at positions 86 and 92 for acquisition of calcium-binding ability. The process of the acquisition of calcium-binding site during evolution of calcium-binding lysozyme is discussed. PMID- 9749922 TI - Modulation of stability properties of bovine trypsin after in vitro structural changes with a variety of chemical modifiers. AB - Controlled chemical modification of enzymes, targeting groups not involved in the active site, can lead to modified catalysts that are intrinsically more efficient and resistant to heat and denaturing agents. Bovine pancreatic trypsin was covalently modified up to 75-85% with monomeric glutaraldehyde (MGA), polymeric glutaraldehyde (PGA), oxidized sucrose and oxidized sucrose polymers (OSP 70 and OSP 400). Virtually no loss in activity occurred upon modification. Temperature optima of trypsin shifts from 45-76 degrees C and T50 from 54-76 degrees C for the best modified sample made with OSP. The efficiency of the modifiers in stabilization was ranked in the order: OSP 400-T > OSP 70-T > PGA-T > MGA-T > Sucrose-T. Half-life of modified enzymes also followed the same trend. Both stabilization factor and t1/2 decreased with increasing temperatures. The free energy of activation for inactivation delta(deltaG*) varies from 12-20 kJ/mol and the activation enthalpy delta(deltaH*) of the modified trypsin by 80-120 kJ/mol indicating stabilization. Inactivation of modified trypsin by urea is less noticeable. The character of the two-step inactivation process of trypsin changes with the degree of stabilization in that the duration of phase I one increased noticeably as stabilization increases. Native trypsin fluoresces less intensely showing a red shift under the influence of denaturation. Such a fluorescence change is not so obvious for the modified enzymes indicating conformational stability acquired by modification. PMID- 9749923 TI - Retardation of thermal and urea induced inactivation of alpha-chymotrypsin by modification with carbohydrate polymers. AB - Modification of enzymes by means of covalent coupling using soluble polymers results in enzymes which retain high biological activity and display resistance to denaturants, high temperature and chaotropic agents. Alpha-chymotrypsin, which has a potential for use in industrial applications, was covalently modified by reductive alkylation using polymeric sucrose (OSP, molecular weight 70 and 400 kDa), dextran (73 and 250 kDa) and carboxymethyl cellulose (CMC, approximately 12 kDa). The derivatives retained around 50-80% activity depending on the polymer used and the extent of modification. At the same time, they displayed better thermotolerance than their native counterpart with 4-14 degrees C higher T50 values. During thermal inactivation, both the native and modified enzymes showed biphasic inactivation kinetics. Half-life of modified enzymes were 2-66-fold greater for the first phase and 5-250-fold greater than the native for the second phase of inactivation. The activation free energy of inactivation of alpha chymotrypsin coupled to polymeric sucrose (400 kDa) was 112.85 kJ/mol for the first phase and 114.71 kJ/mol for the second phase, whereas in the case of the native enzyme, the value for the first phase was 101.55 kJ/mol and 103.42 kJ/mol for the second phase. The activation free energy of inactivation (deltaG*), as well as the activation enthalpy values (deltaH*) of all the modified enzymes were greater than those of the native enzyme, which is an indication of stabilization of the protein and a retardation of inactivation that is usually accompanied by unfolding under thermal and chemical stress. The stability of modified alpha chymotrypsin is in the following order: OSP 400-C > OSP 70-C > CMC-C > Dextran 73 C = Dextran 250-C. PMID- 9749924 TI - Effects of the length of a glycine linker connecting the N-and C-termini of a circularly permuted dihydrofolate reductase. AB - An important consideration in the construction of active and stable circularly permuted proteins is the connective sequence that links the native N- and C termini. For this reason, various lengths of polyglycine linkers (two, three, four, five and six glycines) were employed to connect the original N- and C termini of a circularly permuted construct of Escherichia coli dihydrofolate reductase (DHFR) in which the new N-terminus was Met16. Examination of the circular dichroism (CD) spectra, gel-filtration chromatography elution profiles, urea-induced unfolding properties and enzyme kinetics revealed that, among the linkers tested, a linker length of five glycines was the most favorable. The Vmax of the circularly permuted variant with a five glycine linker (cpM16G5) was about 20% that of wild-type DHFR, although far UV CD spectra, gel filtration elution time, conformational stability and Km for the substrate dihydrofolate and Kd for the coenzyme NADPH were comparable in the two proteins. Another circularly permuted DHFR with a five glycine linker in which a new N-terminus was created at Leu24 (cpL24G5) was also constructed and assayed. The Vmax of cpL24G5 was almost the same as the wild-type, presumably due to the optimization of the glycine linker. The improved activity of the Leu24 permutant is probably due to the disruption of a catalytically important structure, the M20 loop, in the Met16 permutant. PMID- 9749925 TI - Binding of hirudin to meizothrombin. AB - Prothrombin (coagulation factor II) is the inactive precursor molecule of thrombin (coagulation factor IIa). Proteolytic cleavage of the peptide bond Arg320-Ile321 converts prothrombin into the two-chain thrombin precursor meizothrombin. Meizothrombin hydrolyses peptidyl substrates, but cleavage of fibrinogen is poor. Unfortunately, meizothrombin exhibits a significant autocatalytic activity and thus is not structurally stable in solution. Hirudin, the 65-residue peptide anticoagulant from the salivary gland of the European leech Hirudo medicinalis, is a highly specific and effective thrombin inhibitor. To study the interactions of meizothrombin and hirudin, recombinant prothrombin with active site Asp419 replaced by Asn (D419N-prothrombin) was produced in CHO cells and transformed into D419N-meizothrombin in vitro. D419N-meizothrombin exhibited no proteolytic and autocatalytic activity. D419N-meizothrombin was affinity purified at an immobilized C-terminal hirudin-derived peptide demonstrating the presence and activity of the anion binding exosite. D419N meizothrombin exhibited binding activity to hirudin immobilized at the solid phase in an ELISA. Incubation of D419N-meizothrombin with hirudin resulted in a significant increase of intrinsic fluorescence. Fluorescence titration of D419N meizothrombin with hirudin produced a sharp break in the titration curve at the molar equivalence point and a total fluorescence enhancement of 24%. However, the titration curve did not reflect a simple binding mechanism. Incubation of D419N meizothrombin with fibrinopeptide A and C-terminal hirudin peptide 54-65 did not change fluorescence emission. Trp468 located in the gamma-loop of thrombin was replaced by Phe in the double-mutant D419N/W468F-thrombin. Similar to D419N thrombin and D419N-meizothrombin, formation of the D419N/W468F-thrombin/hirudin complex resulted a significant increase in intrinsic fluorescence. Apparently, the binding of hirudin induces similar structural changes in both meizothrombin and thrombin. The structural change does not involve the flexible gamma-loop. The results suggest that meizothrombin binds hirudin similar to thrombin. PMID- 9749926 TI - Is nitric oxide involved in 5-HT3 receptor-mediated neurogenic relaxation of guinea pig proximal colon? AB - The relaxations mediated by the activation of 5-HT receptors in the guinea pig proximal colon were investigated. Longitudinal strips were cut from the colon segment and placed into the bath. In the presence of atropine (0.2 microM), the relaxations were evoked by adding increasing concentrations of 5-HT (1-100 microM). Noncumulative concentration-response curves were established in the absence and presence of either 5-HT or nitric oxide synthase (NOS) antagonists. Selective 5-HT3 antagonists tropisetron (10 and 100 nM) and ondansetron (1 microM) inhibited the relaxations and shifted the concentration-response curves to the right. Similar effects were observed in the presence of the NOS inhibitor N(G)-nitro-L-arginine (3.2, 10, 32 microM) and partly reversed with L-arginine (100, 320 microM). N(G)-nitro-D-arginine, serving as a negative control, was ineffective. The relaxations were further inhibited in the presence of the soluble guanylate cyclase blocker methylene blue (10 microM) or NO scavenger hemoglobin (32 microM). These results suggest that the 5-HT3 receptor plays a role in neurogenic relaxations of guinea pig proximal colon, which are at least partly mediated via release of NO from nerve endings. PMID- 9749927 TI - Relationship between muscarinic autoinhibition and the inhibitory effect of morphine on acetylcholine release from myenteric plexus of guinea pig ileum. AB - The relationship between muscarinic autoinhibition and the inhibitory effect of morphine on acetylcholine (ACh) release was investigated in a longitudinal muscle with myenteric plexus (LMMP) preparation of guinea pig ileum. Morphine (10 microM) inhibited spontaneous and evoked ACh release by electrical field stimulation (EFS) at 1 Hz but not at 10 Hz. Atropine (1 microM) did not affect the resting ACh release, but it significantly increased EFS-evoked release, suggesting activation of muscarinic autoreceptors by ACh released during EFS. Only when the autoinhibition was weakened by atropine, morphine exhibited an inhibitory effect on the EFS-evoked release at 10 Hz. Bethanechol (300 microM) inhibited the EFS-evoked release at 1 Hz but not 10 Hz, suggesting that muscarinic autoreceptors are partially or almost fully activated at 1 or 10 Hz stimulation, respectively. After bethanechol treatment, morphine did not exhibit its inhibitory effect on the EFS-evoked release at 1 Hz. Naloxone (1 microM) increased spontaneous and EFS-evoked ACh release at 1 Hz but not at 10 Hz. Following treatment with atropine, naloxone also increased ACh release at 10-Hz stimulation. These results suggest that morphine and an endogenous opioid inhibit ACh release from LMMP preparations when muscarinic autoinhibition mechanism does not fully work. This inhibitory effect of morphine is discussed in relation to the calcium sensitivity of the preparations in ACh release. PMID- 9749928 TI - Relationship between inhibitory effect of endogenous opioid via mu-receptors and muscarinic autoinhibition in acetylcholine release from myenteric plexus of guinea pig ileum. AB - Relationship between activation of opioid receptors and muscarinic autoinhibition in acetylcholine (ACh) release from the myenteric plexus was studied in longitudinal muscle myenteric plexus (LMMP) preparations of guinea pig ileum. A mu-receptor agonist, [D-Ala2, N-Me-Phe4, Gly5-ol] enkephalin (DAMGO), at a concentration of 1 microM inhibited the ACh release evoked by electrical field stimulation (EFS) at 1 Hz but not at 10 Hz. After the muscarinic autoreceptors were blocked with atropine (1 microM), DAMGO inhibited EFS-evoked ACh release also at 10 Hz. After the autoreceptors were potently activated with muscarine (200 microM), the inhibitory effect of DAMGO at 1 Hz was abolished. A kappa receptor agonist, U-50,488, at 1 microM inhibited the EFS-evoked ACh release both at 1 and 10 Hz. U-50,488 inhibited ACh release regardless of the presence of atropine or muscarine. A delta-agonist, enkephalin [D-PEN2.5] (PDPDE), did not show any significant effect. On the other hand, a selective mu-receptor antagonist, cyprodime, increased ACh release evoked by EFS at 1 Hz, but not at 10 Hz. After the autoreceptors were blocked, cyprodime increased EFS-evoked ACh release also at 10 Hz. The selective kappa-receptor antagonist, nor binaltorphimine, did not affect ACh release in the absence or presence of atropine. The results suggest that endogenous opioid(s) inhibits ACh release by activating mu-, but not kappa- and delta-receptors in the LMMP of guinea pig ileum and that the inhibitory effect of endogenous opioid(s) in the ACh release is important when muscarinic autoinhibition mechanism does not fully work. PMID- 9749929 TI - Cold exposure enhances nitroxidergic nerve-mediated vasodilatation in canine nasal mucosa. AB - We have previously reported that there is non-adrenergic, non-cholinergic (NANC) innervation in canine nasal mucosa and that the relaxation response to electrical stimulation of the NANC nerve is mainly mediated by nitric oxide (NO). In the present study, we examined the effect of cold exposure (24 degrees C) on nitroxidergic nerve-mediated vasodilatation in isolated canine nasal mucosa. Nasal mucosa strips, prepared from canine nasal septum and moderately precontracted with methoxamine in the presence of atropine and guanethidine, relaxed in response to transmural electrical stimulation (square pulses of 0.5 msec duration, at 5 Hz and 25 V). The degree of relaxation at 24 degrees C (55.4+/-13.2% of methoxamine-induced contraction, mean+/-S.D., n=6) was significantly greater than that at 34 degrees C (33.8+/-8.6%, n=6). This phenomenon was reversible. In contrast, the magnitude of relaxation responses to an NO donor (sodium nitroprusside of 0.1 and 1 microM) remained unchanged by cold exposure. These results suggest that the release of NO from the nitroxidergic nerve endings is augmented by cold exposure and, thus, vasodilatation of the nasal blood vessel is enhanced, thereby contributing to the swelling of the nasal mucosa in cold conditions. PMID- 9749930 TI - Irsogladine maleate may preserve gastric mucosal hydrophobicity against ethanol in phospholipids independent way in rats. AB - Irsogladine maleate (IM) has been used as a mucosal protective agent, whose action is partially explained as enhancement of mucosal blood flow, increase of cellular cyclic AMP and facilitation of gap-junctional intercellular communication. Effect of IM on rat gastric mucosal hydrophobicity, one of the mucosal barrier properties, was investigated, in comparison with that of 16,16 dimethyl prostaglandin E2 (dmPGE2). IM alone had no effect on mucosal hydrophobicity and mucosal phospholipids content. dmPGE2 alone did not change mucosal hydrophobicity significantly, but remarkably increased mucosal surface active phospholipids. Intragastric administration of absolute ethanol significantly decreased gastric mucosal hydrophobicity and mucosal phospholipids content. IM could prevent the decrease in mucosal hydrophobicity by ethanol, maintaining the surface mucus gel layer and mucosal surface phospholipids almost as non-damaged control levels, whereas dmPGE2 also prevented the decrease in mucosal hydrophobicity by ethanol, with the surface epithelium being partially exfoliated and mucosal surface-active phospholipids showing remarkable enhancement. These results suggest that IM may preserve gastric mucosal hydrophobicity against ethanol, not through enhancement of mucosal phospholipids content like prostaglandin, but possibly through its reported stabilization action to the epithelial cell lining, which may preserve the surface epithelium with the mucous gel layer containing surface-active phospholipids, a possible origin of mucosal hydrophobicity. PMID- 9749931 TI - Bremazocine recognizes the difference in four amino acid residues to discriminate between a nociceptin/orphanin FQ receptor and opioid receptors. AB - We investigated the molecular basis of the discrimination between nociceptin/orphanin FQ receptor (NociR) and opioid receptors (OPRs) by bremazocine, a non-type-selective opioid ligand. Construction of several chimeric receptors between NociR and kappa-opioid receptor (KOPR) and mutant NociRs followed by binding experiments with [3H]bremazocine showed that the mutation of only four amino acid residues of NociR, Ala216, Val279, Gln280 and Val281, to the amino acid residues located at the corresponding position of KOPR, Lys227, Ile290, His291 and Ile292, made it possible for the resultant mutant NociR to bind bremazocine with high affinity. Considering that these four amino acid residues are conserved among mu-, delta- and kappa-OPRs, the present result suggests that bremazocine recognizes the difference in these four amino acid residues to discriminate between NociR and OPRs. PMID- 9749932 TI - Endothelin receptors in testosterone-induced prostatic hypertrophy in rats. AB - Endothelin receptors were characterized in rat prostate and potential modification of these receptors was investigated in prostatic hypertrophy induced by testosterone. Both ET(A) and ET(B) endothelin receptor mRNA were detected in rat prostate, whereas binding experiments show the presence of only ET(A) receptors. Testosterone administration produced a 75% increase in prostate weight. Although the density of prostatic endothelin receptors was decreased from 348 +/- 75.0 fmol/mg protein in control rats to 252 +/- 39.9 fmol/mg protein in testosterone-treated animals, the total amount of receptors per prostate was unchanged. The steady-state level of ET(A)- and ET(B)-receptor mRNA was not altered by testosterone treatment. These results suggest that endothelin receptors are not affected in prostatic hypertrophy induced by testosterone. PMID- 9749933 TI - 17Beta-estradiol alters isoproterenol-induced relaxation in rat aortic rings. AB - Female Wistar rats were treated with 17beta-estradiol (E2) (10 microg, s.c.) or with sesame oil for 3 days. The relaxation induced by isoproterenol (10(-9)-3 x 10(-6) M) in aortae precontracted with norepinephrine was significantly suppressed in aortae from E2-treated rats compared with the relaxation in those from control rats. N(G)-Nitro-L-arginine, a nitric oxide synthase inhibitor, inhibited isoproterenol-induced relaxation in aortae from both E2-treated and control rats. Metyrapone, a cytochrome P-450 monooxygenase inhibitor, inhibited it in aortae from control rats, but metyrapone enhanced the maximum relaxation in aortae from E2-treated rats. These results suggest that E2 modulates isoproterenol-induced vasodilation through nitric oxide and cytochrome P-450 dependent metabolites. PMID- 9749934 TI - Intrahepatic flow disturbance as detected by in vivo acridine orange staining in endothelin-1-treated and cirrhotic rats. AB - Intrahepatic flow in rats was examined by systemic acridine orange (AO) labeling and succeeding microscopic observations of post-fixed livers. In controls, all periportal areas were stained evenly throughout the liver mass, and the staining often branched between the liver lobules in a acinous fashion. Intraportal endothelin-1 infusion caused a marked heterogenous staining; strong staining around large portal vein branches in the central portion of the liver, but much less in the periphery. Cirrhotic livers exhibited overall weaker staining intensity with no accentuation in periportal areas and restricted radial staining within pseudo-lobules. AO infusion may help detect hepatic flow disturbance in vivo. PMID- 9749935 TI - Flight crew fatigue I: objectives and methods. AB - In 1980, NASA-Ames Research Center, Moffett Field, CA, initiated a program to assess flight crew fatigue, determine its potential operational consequences, and provide practical countermeasure suggestions. To assess the extent of the problem, crewmembers were monitored before, during, and after commercial short haul (fixed-wing and helicopter aircraft), overnight cargo, and long-haul operations. A total of 197 volunteers were studied on 94 trip patterns with 1299 flight segments and 2046 h of flying time. The present paper outlines the program and describes the common methodology used in these studies, which are then presented in detail in the four subsequent papers. The sixth paper offers a synthesis of this work, reviewing the major causes of flight crew fatigue and making specific suggestions about ways to manage it in different operations. PMID- 9749936 TI - Flight crew fatigue II: short-haul fixed-wing air transport operations. AB - We monitored 74 crewmembers before, during, and after 3-4-d commercial short-haul trips crossing no more than one time zone per 24 h. The average duty day lasted 10.6 duty hours, with 4.5 flight hours and 5.5 flights. On trips, crewmembers slept less, woke earlier, and reported having more difficulty falling asleep, with lighter, less restful sleep than pretrip. The consumption of caffeine, alcohol, and snacks increased on trip days, as did reports of headaches, congested nose, and back pain. The study suggests the following ways of reducing fatigue during these operations: base the duration of rest periods on duty hours as well as flight hours; avoid scheduling rest periods progressively earlier across a trip; minimize early duty report times; and inform crewmembers about strategic use of caffeine and alternatives to alcohol for relaxing before sleep. PMID- 9749937 TI - Flight crew fatigue III: North Sea helicopter air transport operations. AB - We studied 32 helicopter pilots before, during, and after 4-5 d trips from Aberdeen, Scotland, to service North Sea oil rigs. On duty days, subjects awoke 1.5 h earlier than pretrip or posttrip, after having slept nearly an hour less. Subjective fatigue was greater posttrip than pretrip. By the end of trip days, fatigue was greater and mood more negative than by the end of pretrip days. During trips, daily caffeine consumption increased 42%, reports of headache doubled, reports of back pain increased 12-fold, and reports of burning eyes quadrupled. In the cockpits studied, thermal discomfort and high vibration levels were common. Subjective workload during preflight, taxi, climb, and cruise was related to the crewmembers' ratings of the quality of the aircraft systems. During descent and approach, workload was affected by weather at the landing site. During landing, it was influenced by the quality of the landing site and air traffic control. Beginning duty later, and greater attention to aircraft comfort and maintenance, should reduce fatigue in these operations. PMID- 9749938 TI - Flight crew fatigue IV: overnight cargo operations. AB - We monitored 34 B-727 crewmembers before, during, and after 8-d commercial overnight cargo trips crossing no more than one time zone per 24 h. Daytime sleep episodes were 41% shorter and were rated as poorer than nighttime sleep episodes. When the layover was long enough, crewmembers usually slept again in the evening before going back on night duty. Nevertheless, the total sleep per 24 h on duty days averaged 1.2 h less than pretrip. The circadian temperature rhythm did not adapt completely to night duty, delaying by about 3 h. Self-rated fatigue was highest around the time of the temperature minimum, which occurred near the end of the nighttime duty period. On trip days, crewmembers ate more snacks and there was a marked increase in reports of headaches, congested noses, and burning eyes. Comparisons with daytime short-haul operations confirm that a daytime rest period does not represent the same sleep opportunity as a nighttime rest period of the same duration. We examine regulatory and scheduling options, and personal countermeasure strategies, that could help to reduce sleep loss during overnight cargo operations. PMID- 9749939 TI - Flight crew fatigue V: long-haul air transport operations. AB - We monitored 32 flight crewmembers before, during, and after 4-9 d commercial long-haul trips crossing up to 8 time zones per 24 h. The average duty day lasted 9.8 h, and the average layover 24.8 h. Layover sleep episodes averaged 105 min shorter than pretrip sleep episodes. However, in two-thirds of layovers, crewmembers slept twice so that their total sleep per 24 h on trips averaged 49 min less than pretrip. Greater sleep loss was associated with nighttime flights than with daytime flights. The organization of layover sleep depended on prior flight direction, local time, and the circadian cycle. The circadian temperature rhythm did not synchronize to the erratic environmental time cues. Consequently, the circadian low point in alertness and performance sometimes occurred in flight. On trip days, by comparison with pretrip, crewmembers reported higher fatigue and lower activation; drank more caffeine; ate more snacks and fewer meals; and there were marked increases in reports of headaches, congested nose, and back pain. Scheduling strategies and countermeasures to improve layover sleep, cockpit alertness, and performance, are discussed. PMID- 9749940 TI - Flight crew fatigue VI: a synthesis. AB - Sleep, circadian rhythms, subjective fatigue, mood, nutrition, and physical symptoms were monitored in flight crews before, during, and after scheduled commercial operations. Duty-related changes in these measures were examined in four different types of air transport: short-haul fixed-wing; short-haul helicopter; domestic overnight cargo; and long-haul. The extent of these changes, and the duty-related and physiological factors contributing to them, are compared among the different operations. During all operations, the level of sleep loss was such that the majority of crewmembers would be expected to have become increasingly sleepy across trip days, with some experiencing performance decrements. In addition, during overnight cargo and long-haul operations, crewmembers were sometimes flying aircraft during the circadian low point in alertness and performance. Specific recommendations for reducing flight crew fatigue are offered for each operating environment. PMID- 9749941 TI - Intracellular trafficking of tropoelastin. AB - Elastin is secreted as soluble tropoelastin monomers which are then cross-linked in the presence of extracellular microfibrils to form insoluble elastic fibers. Although the secretion of tropoelastin is thought to be mediated and targeted by an intracellular chaperone complex, the intracellular route taken by this protein and the role of such a chaperone complex remain undefined. In the present study, the specific pathway of tropoelastin secretion was investigated in fetal bovine chondrocytes and ligamentum nuchae fibroblasts by immunofluorescence staining and immunoprecipitation of tropoelastin following treatment with secretion-disrupting agents. In untreated cells, tropoelastin is secreted in approximately 30 min. In both cell types, brefeldin A and monensin inhibited secretion of tropoelastin and caused an intracellular accumulation of the protein in the fused ER/Golgi compartment or in the Golgi stacks, respectively. Incubations of longer than 1 h in the presence of brefeldin A result in eventual degradation of tropoelastin in the ER/Golgi compartment (Davis and Mecham, 1996). In contrast, the tropoelastin trapped in the Golgi as a result of monensin treatment steadily accumulated. Agents that elevate intracellular pH, such as ammonium chloride and chloroquine, also caused an intracellular accumulation of tropoelastin which appeared by immunofluorescence staining to be localized in secretory vesicles and/or endosomes. Since weak bases and ionophores alter the morphology of vacuolar compartments, the effect of bafilomycin A1 on tropoelastin secretion was also investigated. This vacuolar H+-ATPase inhibitor prevents acidification of the trans-Golgi network and endosomal compartments without disrupting intracellular organelle formation. When the elastogenic cells were treated with bafilomycin A1, tropoelastin secretion was diminished and an intracellular accumulation of tropoelastin was detected in the trans-Golgi network and small secretory vesicles. These results suggest that tropoelastin may be diverted from the constitutive pathway after exiting the Golgi and instead targeted to an acidic compartment prior to transport to the cell surface. The identity and role of such a compartment in the sorting and/or trafficking of tropoelastin has yet to be determined. PMID- 9749942 TI - Structural analysis of proteoglycans synthesized by mineralizing bone cells in vitro in the presence of fluoride. AB - This study investigated the biochemical structure of proteoglycans synthesized during matrix maturation by mineralizing bone cells in vitro, in the presence and absence of fluoride. Bone cells were obtained from rat femur washes and cultured in alpha MEM media supplemented with fetal calf serum, ascorbic acid, beta glycerophosphate and dexamethasone. Cells were characterized as osteoblast-like by the expression of alkaline phosphatase activity and the synthesis of collagen type I and osteocalcin. Fluoride, present in the culture media at concentrations of 10(-5) M or 10(-7) M, had negligible effect on cell viability. However, calcium deposition was increased in cell cultures incubated in the presence of fluoride. Proteoglycans were extracted from the extracellular matrix with 4 M guanidinium chloride and purified by anion exchange chromatography. Biochemical analysis identified the presence of the small leucine rich proteoglycan, decorin and biglycan, in addition to degradation products relating to the larger chondroitin sulphate protoeglycan, versican. Fluoride had little effect on the size or amino acid composition of the protein core, but resulted in significant alterations to the GAG chains, including a dramatic reduction in chain length, reduction in sulphation and decrease in the proportion of dermatan sulphate compared to chondroitin sulphate. The influence of fluoride on proteoglycan structure synthesized by mineralizing bone cells provides valuable information, indicating specific roles for dermatan sulphate and chondroitin sulphate proteoglycans. The results suggested that fluoride affected the post translational assembly of the GAG chains which may be an influential factor in the mineralization process. PMID- 9749943 TI - Characterization of cartilage oligomeric matrix protein (COMP) in human normal and pseudoachondroplasia musculoskeletal tissues. AB - Cartilage oligomeric matrix protein (COMP), the fifth member of the thrombospondin gene family, is an extracellular matrix calcium-binding protein. The importance of COMP is underscored by the finding that mutations in COMP cause the human dwarfing condition, pseudoachondroplasia (PSACH). Here, we report the results of human tissue distribution and cell secretion studies of human COMP. COMP is expressed and secreted by cultured monolayer chondrocyte, tendon and ligament cells, and COMP secretion is not restricted to a differentiated chondrocyte phenotype. Whereas COMP is retained in the endoplasmic reticulum that accumulates within PSACH chondrocytes in vivo, COMP is not retained intracellularly in the dedifferentiated PSACH chondrocytes in cultures. These results lend further support to the hypothesis that retention of COMP is related to the terminal PSACH chondrocyte phenotype, processing of proteins related to extracellular matrix formation, and maintenance in cartilage. PMID- 9749944 TI - Expression of mRNA for type IV collagen alpha1, alpha5 and alpha6 chains by cultured dermal fibroblasts from patients with X-linked Alport syndrome. AB - COL4A5 mutations causing X-linked Alport syndrome (XLAS) are frequently associated with absence of the alpha3, alpha4,alpha5 and alpha6 chains of type IV collagen from basement membranes and increased amounts of the alpha1(IV) and alpha2(IV) chains in glomerular basement membrane. Although many COL4A5 mutations have been described in XLAS, the mechanisms by which these mutations influence the basement membrane appearance of chains other than alpha5(IV) remain poorly understood. In this study, we used dermal fibroblasts from eight normal individuals and nine males with XLAS to test the hypotheses that COL4A5 mutations increase transcription of COL4A1 and suppress transcription of COL4A6. Ribonuclease protection assays revealed that alpha1(IV), alpha5(IV) and alpha6(IV) transcripts were expressed in cultures of dermal fibroblasts. The mRNA levels for alpha1(IV) in eight of nine patients with XLAS were not increased compared to controls; one patient with a large COL4A5 deletion showed significant elevation of alpha1(IV) mRNA levels. No differences in steady-state mRNA levels for alpha6(IV) were found when XLAS fibroblasts were compared with controls, even though little or no alpha6(IV) protein was detectable at the dermal-epidermal junction by immunofluorescence study. This finding suggests that post transcriptional events account for the absence of alpha6(IV) in the Alport dermal epidermal junction. PMID- 9749945 TI - Collagenase-1, stromelysin-1 and 92 kDa gelatinase are associated with tumor necrosis factor-alpha induced morphological change of human endothelial cells in vitro. AB - Recently, we have shown that the tumor necrosis factor-alpha (TNF-alpha)-induced morphological change of EA.hy 926 human endothelial cells is associated with a decrease in the net synthesis of two proteoglycans (PGs), biglycan and syndecan 1, both of which have been suggested to play a role in cell adhesion. Here we have examined whether this phenotypic modulation of EA.hy 926 cells also involves altered expression of matrix metalloproteinases (MMPs) or their tissue inhibitors (TIMPs). We demonstrate that, when forming cobblestone-like monolayer cultures, these cells express and synthesize collagenase-1 (MMP-1), stromelysin-1 (MMP-3) and 72 kDa (MMP-2) and 92 kDa (MMP-9) gelatinases, all of which have previously been found in either normal or pathological human vascular wall. EA.hy 926 cells also express membrane-typel MMP (MT1-MMP), but not matrilysin (MMP-7) and collagenase-3 (MMP-13). As regards TIMPs, we show that these cells express TIMP-1 and TIMP-2, but not TIMP-3 or TIMP-4. Exposure of the cells to TNF-alpha changed the cell morphology from a polygonal shape into a spindle shape and also increased the mRNA levels of MMP-1, MMP-3 and MMP-9, but slightly decreased the MMP-2 mRNA level. No change at the mRNA level of MT1-MMP was observed. Similarly to unstimulated cultures, no mRNA for MMP-7 or MMP-13 was detected in the TNF alpha treated cultures. TNF-alpha had no effect on the TIMP-1 and TIMP-2 mRNA levels and did not induce TIMP-3 or TIMP-4 expression. Gelatin zymography and Western blot analysis revealed that the increase observed at the mRNA level of MMP-3 and MMP-9 was similar to that of their net protein level; furthermore, the active form of MMP-1 was induced. Our results indicate that the TNF-alpha-induced morphological change of EA.hy 926 cells is associated not only with specific changes in the expression of PGs by the cells, but also with specific changes in the expression of MMPs. PMID- 9749947 TI - Production of cartilage collagens during metaphyseal bone healing in the mouse. AB - Small defects of unfractured bone are believed to heal without a cartilaginous intermediate. We have determined the extent of cartilage production in an experimental model of metaphyseal bone repair involving defects in both cortical and cancellous bone, but no fracture. Northern analyses revealed the presence of mRNAs for type X and II collagens in the repair tissue. Immunohistology confirmed subperiosteal deposition of both collagen types adjacent to the defect. While the mRNAs for the two collagen types peaked by one week of defect healing, immunodetectable type X collagen was not observed until the second week. The data suggest that reactivity of periosteum and activation of chondrogenesis and subsequent endochondral ossification programs are involved in murine bone repair regardless of defect type. PMID- 9749946 TI - Tenascin-C expression in human epidermal keratinocytes is regulated by inflammatory cytokines and a stress response pathway. AB - Recently we showed that human epidermal keratinocytes express the extracellular matrix protein tenascin-C (TN-C) during wound healing, but not in normal adult skin. To gain further insight into the regulation of epidermal TN-C expression, we tested the effect of various stimuli on TN-C expression by cultured keratinocytes. Our results indicate that IL-4 is a very strong inducer of TN-C protein and mRNA expression in normal keratinocytes. Furthermore, TNFalpha and IFNgamma moderately increased TN-C expression. No other cytokines and growth factors that we tested, including various factors that stimulate TN-C expression in mesenchymal cells, significantly affected TN-C secretion by cultured keratinocytes. The regulation of TN-C expression in keratinocytes is distinct from that of fibronectin, since IL-4 and IFNgamma did not affect fibronectin expression in our experiments, and TNFalpha only slightly increased fibronectin levels. To investigate the role of cellular stress response pathways that can be activated by TNFalpha in the regulation of TN-C expression, we tested the effect of different inhibitors and an activator of these intracellular signalling cascades. The results show that the p38 MAP-kinase pathway is not involved in TNFalpha-induced TN-C expression in cultured keratinocytes. Activation of the JNK/SAPK-1 pathway by the addition of sphingomyelinase resulted in a dose dependent increase of TN-C expression. TN-C expression by squamous carcinoma cell lines was differentially affected by the cytokines that stimulated TN-C expression in normal keratinocytes: TNFalpha again increased TN-C secretion, but IL-4 and IFNgamma had little effect. We conclude that there are distinct regulation mechanisms for TN-C expression in normal keratinocytes, tumor-derived keratinocytes and mesenchymal cells. The observation that TN-C is abundant in inflamed skin is a strong indication that inflammatory cytokines such as IL-4, TNFalpha and IFNgamma could also be involved in the regulation of epidermal TN-C expression in vivo. PMID- 9749948 TI - Carotid body I1-imidazoline receptors: binding, visualization and modulatory function. AB - The carotid body is influenced by many neurotransmitter receptors. A novel receptor specific for imidazolines has been implicated in cardiorespiratory regulation in the brain. To test for both I1-imidazoline and alpha2-adrenergic receptors, which also recognize imidazolines, specific [125I]p-iodoclonidine binding to carotid body membranes was characterized. The specific alpha2-agents epinephrine (100 microM) or SK&F 86466 (10 microM) inhibited only a portion of specific [125I]p-iodoclonidine binding in both cat and rabbit carotid bodies, indicating the presence of I1-imidazoline as well as alpha2-adrenergic sites. The distribution of [125I]p-iodoclonidine binding sites was visualized autoradiographically. The cat carotid body was intensely labeled by [125I]p iodoclonidine, with both I1-imidazoline and alpha2-adrenergic sites expressed. The relevance of I1-imidazoline receptors in modulation of chemosensory discharge was determined in seven cats after alpha2-adrenergic blockade. Clonidine (100 microg/kg) facilitated chemosensory activity particularly under hypoxia. We conclude that I1-imidazoline receptors are expressed within the carotid body and may potentiate chemosensory discharge, in contrast to the inhibitory action of alpha2-adrenergic receptors. PMID- 9749949 TI - Effects of volatile anesthetics on vagal C-fiber activities and their reflexes in anesthetized dogs. AB - Effects of halothane, enflurane, isoflurane, and sevoflurane on vagal capsaicin (CAPS)-sensitive C-fibers were elucidated in anesthetized dogs. The CAPS sensitive C-fibers were significantly stimulated by all volatile anesthetics with a significantly greater response to halothane than with sevoflurane. A significant increase in respiratory frequency (fR) and a significant decrease in tidal volume (VT) were observed with halothane and isoflurane, and a significant increase in fR was observed with sevoflurane. In contrast, a significant decrease in fR was induced by enflurane. The tachypnea induced by halothane, isoflurane, and sevoflurane was significantly reduced or no longer observed after perineural CAPS-treatment or bilateral vagotomy, whereas the slowing of respiration observed with enflurane was not affected by either of these treatments. These results suggest that vagal C-fibers play an important role in the reflex tachypnea that occurs with halothane, isoflurane, and sevoflurane. PMID- 9749950 TI - Chemotransduction by carotid body chemoreceptors is dependent on bicarbonate currents. AB - Previous studies have demonstrated that bicarbonate enhances the speed and magnitude of the carotid body chemoreceptor response to hypoxia. We hypothesized that this enhancement is associated with enhanced hypoxia-induced catecholamine (CAT) secretion from glomus cells. Single-fiber nerve activity and free tissue catecholamine (carbon fiber microvoltammetry) were measured in rat carotid body, in vitro. The peak CAT and nerve responses during 1 min anoxia were larger in the presence of bicarbonate than in its absence (peak CAT: 16.7 +/- 2.7 vs. 5.1 +/- 1.1 microM; peak nerve: 28.2 +/- 1.6 vs. 16.7 +/- 1.4 Hz). Bicarbonate particularly enhanced the responses to moderate hypoxia (PO2 approximately 80 Torr) which caused no secretion or increased nerve activity in the absence of bicarbonate, but caused significant stimulation in the presence of bicarbonate (peak nerve = 15.2 Hz; peak CAT = 8.6 microM). The bicarbonate effect was not due to alterations in intracellular pH since it was not blocked by exchanger blockers (DIDS) or mimicked by acidification of the medium. However, anion channel blockade by 9-AC or DPC reduced anoxia-induced CAT secretion in the presence of bicarbonate. We conclude that bicarbonate greatly enhances stimulus/secretion coupling in glomus cells, probably through modulation of an anion current carried by bicarbonate. PMID- 9749951 TI - Modulation of the hypoxic ventilatory response by Ca2+-dependent and Ca2+ independent protein kinase C in the dorsocaudal brainstem of conscious rats. AB - Protein kinase C (PKC) activation in the nucleus tractus solitarii (NTS) is critical for mounting an appropriate hypoxic ventilatory response (HVR). Furthermore, hypoxia elicits translocation of both Ca2+-dependent and Ca2+ independent PKC isoforms in the NTS. However, the relative functional contribution of such PKC isoforms in mediating HVR is unclear. To study these issues, chronically instrumented adult Sprague-Dawley rats underwent hypoxic challenges (10% O2 balance in N2) following dorsocaudal brainstem microinjections of the selective Ca2+-dependent PKC inhibitor Go 6976 (10 mmol in 1 microl). Compared with vehicle, Go 6976 did not modify normoxic ventilation but maximally attenuated HVR by 38.4 +/- 6.7% (n = 9; P < 0.01), with similar contributions from tidal volume and respiratory frequency. In seven additional animals, when the non Ca2+-selective PKC blocker BIM I was concurrently microinjected with Go 6976, further reductions in peak ventilatory responses to hypoxia occurred (P < 0.04). When BIM V, the inactive analog, was microinjected with Go 6976, the magnitude of HVR attenuation was unchanged (n = 6; Go 6976 vs. Go 6976 + BIM V: P = NS). We conclude that in the dorsocaudal brainstem, PKC-mediated components of HVR involve activation of both Ca2+-dependent and Ca2+-independent PKC isoforms. PMID- 9749952 TI - Ventilatory dynamics of transient arousal in patients with obstructive sleep apnea. AB - The hyperpnea that accompanies arousal at the end of obstructive apnea is believed to be due to the progressive build-up in chemical drive during the apnea and a state-related decrease in upper airway resistance. We postulated the existence of a third component: a state-related transient increase in neural drive to the ventilatory pump muscles. To quantify this contribution, we measured the ventilatory response to arousal (VRA) in eight patients with obstructive sleep apnea (OSA) during continuous positive airway pressure (CPAP) therapy, applied at individually titrated levels. CPAP application reduced total pulmonary resistance (RL) to approximately normal levels, stabilizing ventilation and sleep state. Transient arousal from stage 2 sleep was induced using 5-sec tones (60-90 dB). Mean inspiratory flow increased above control on the second and third post arousal breaths (P < 0.05), with a peak increase of 7.8 +/- 2.9 L/min while the accompanying changes in RL were significant. The time-course of VRA measured in three normal subjects under CPAP was similar to that observed in the OSA patients. However, elimination of CPAP prolonged the VRA time-course. Taken together, these findings demonstrate that: (1) during arousal, the increase in state-related neural respiratory drive is short-lived but not substantial; and (2) the resulting VRA time-course is shaped by the dynamics of the upper airway response to arousal. PMID- 9749953 TI - High-frequency oscillation and centroid frequency of diaphragm EMG during inspiratory loading. AB - Power spectra were derived from the diaphragm electromyogram (EMG) in anesthetized rabbits subjected to inspiratory resistive loading (IRL) with airway pressure swings of 40-60 cm H2O for 20 min to 2 h. Shifts in the centroid frequencies of the power spectra were found to be associated with the appearance of power spectral peaks in the range of 105-140 Hz, termed high-frequency oscillation, or HFO. Such peaks have been described before in phrenic nerve activity and in the diaphragm EMG. However, these peaks have not previously been connected with the shifts in centroid frequency seen during loaded breathing. Although such changes in frequency content have been taken to indicate fatigue in the diaphragm, we find that HFO can also cause a shift in centroid frequency during loaded breathing, an effect whose relation to fatigue has yet to be established. PMID- 9749954 TI - Neuromodulators and hypoxic hypothermia in the rat. AB - This study was designed to assess if opioids or adenosine are involved in the hypometabolism induced by hypoxia in the rat. Accordingly, antagonists such as naloxone (NLX) for opioids or theophylline (THEO) for adenosine were injected into conscious adult rats acutely exposed to either ambient hypoxia (AHx, FIO2: 12%) at ambient temperatures of 26 or 9 degrees C, or to CO hypoxia (COHx, FICO = 0.05%) at an ambient temperature (Ta) of 9 degrees C. Oxygen consumption, ventilation, colonic temperature and shivering were recorded. The results show that with NLX, the degree of hypoxic hypometabolism was reduced with AHx at 26 degrees C and slightly decreased with COHx at 9 degrees C. With THEO, hypoxic hypometabolism was slightly reduced with AHx and COHx at 9 degrees C. The ventilatory response to AHx and COHx was not consistently affected by either NLX or THEO. It is concluded that adenosine and opioids play a minor role, in mediating AHx or COHx hypothermia, especially during cold exposure. PMID- 9749955 TI - Morphometric analysis of postnatal lung development in the tammar wallaby: light microscopy. AB - Postnatal growth of the lung in the tammar wallaby, Macropus eugenii, was investigated using morphometric techniques with light microscopy. Lung volume, parenchymal and non-parenchymal volume densities were measured. Volume densities of parenchymal airspace and tissue and non-parenchymal conducting airways and large blood vessels were determined. Lung volume and all the other parameters that were measured showed a biphasic increase in relation to increase in body mass. All parameters, with the exception of airway volume, increased relatively slowly in relation to increase in mass in the first 70 days after birth, when the pouch young are ectothermic. Between 70 and 180 days, during the period of transition from ectothermy to endothermy, the parameters increased more rapidly, suggesting accelerated lung growth in preparation for the extra metabolic demands associated with the establishment of thermoregulatory control in the pouch young. Specific lung volume in the adult tammar is lower than that of eutherians of equivalent mass, however, the parenchymal volume is relatively high. PMID- 9749956 TI - Changed expression of 9-O-acetyl GD3 (CDw60) in benign and atypical proliferative lesions and carcinomas of the human breast. AB - Expression of gangliosides is affected in various ways by malignant cell transformation. In the present study, we investigated the expression of CDw60, a constituent of O-acetylated disialogangliosides, in benign and atypical proliferative breast diseases, and preinvasive and invasive carcinomas by immunohistochemistry and thin-layer chromatography (TLC). In normal ducts, antibodies to CDw60 (mAb M-T21) reacted to membranes of the Golgi apparatus in the juxtaluminal cell compartment. A similar polarized distribution of Golgi cisterns in epithelial cells was observed in several benign lesions, i.e., fibroadenomas, intraductal papillomas, and gynecomastia. In contrast, blunt duct adenosis and duct hyperplasia exhibited an abnormal cytosolic and cell surface staining, whereas atypical duct hyperplasia showed randomly dispersed immunoreactive Golgi cisterns, indicating loss of epithelial polarity. In mammary carcinomas and in two breast carcinoma cell lines (MCF-7 and EFM-19) the neoplastic cells contained CDw60-immunolabelled Golgi complexes, which were distributed in a disorderly fashion throughout the cytoplasm, thus reflecting a loss of epithelial polarity. Additionally, only well differentiated ductal carcinomas in situ or invasive ductal carcinomas disclosed a strong cell surface labelling, which was absent in lower differentiated carcinomas of the same types. In all carcinomas, the intensity of CDw60 immunostaining decreased with progressing loss of differentiation (grade of dedifferentiation), as demonstrated by staining intensity in paraffin sections and by evaluation of the relative amounts of extracted 9-O-acetyl GD3 by TLC. Our results indicate that abnormal CDw60 expression is already detectable in benign proliferative breast lesions with different risk rates to develop into malignant lesions. Downregulation of CDw60 expression in poorly differentiated invasive carcinomas may be the consequence of loss of cell functions usually associated with poor prognosis. PMID- 9749957 TI - Differential expression of CD14, CD36 and the LDL receptor on human monocyte derived macrophages. A novel cell culture system to study macrophage differentiation and heterogeneity. AB - Macrophages are key players in many aspects of human physiology and disease. It has been hypothesized that in a given microenvironment monocytes differentiate into specific subpopulations with distinct functions. In order to study the role of macrophage heterogeneity in atherogenesis, we established a novel isolation and culture technique for human monocyte-derived macrophages. The present technique does not select for monocyte subpopulations prior to the onset of differentiation. Monocytes were cultured for 2 weeks in the presence of autologous lymphocytes before being plated quantitatively. They differentiated into mature macrophages in terms of morphology, lipid composition, and biological activity. Based on phagocytic activity as well as on the expression of CD14, CD36, and the low-density lipoprotein (LDL) receptor, we have identified macrophage subpopulations that may play distinct roles in atherogenesis. While virtually all adherence-purified monocytes expressed CD14, CD36, and the LDL-R, we characterized three subpopulations of macrophages based on the expression of these antigens: CD36+CD14-LDL-R-(58+/-12%), CD36+CD14+LDL-R+(18+/-5%), the remaining cells being CD36-CD14- LDL-R-. The first two subsets decreased in size during further differentiation (51+/-12% and 8+/-3%, respectively). Our culture technique may also serve as a good model for studying the implications of macrophage heterogeneity in diseases other than atherosclerosis. PMID- 9749958 TI - Tissue and subcellular distribution of mercaptopyruvate sulfurtransferase in the rat: confocal laser fluorescence and immunoelectron microscopic studies combined with biochemical analysis. AB - In our previous study, we found that mercaptopyruvate sulfurtransferase (MST) was evolutionarily related to mitochondrial rhodanese. To elucidate the difference between MST and rhodanese, the tissue, cellular, and subcellular distribution of rat MST was determined biochemically and immunohistochemically by using anti-MST antibody raised in rabbit. In an immunohistochemical study, tetramethyl rhodamine isothiocyanate-conjugated phalloidin against F-actin and fluorescein isothiocyanate-conjugated goat anti-rabbit immunoglobulin as a secondary antibody to the anti-MST antibody were used for double fluorescent staining. They were detected by confocal laser fluorescence microscopy. In the immunoelectron microscopic study of hepatocyte and renal tubular epithelium, a postembedding immunogold method was used. Biochemical studies including western blot analyses of various tissues and subcellular fractions of the liver were also performed. MST was widely distributed in rat tissues but the cellular distribution was found to be different in each tissue. MST was predominantly localized in proximal tubular epithelium in the kidney, pericentral hepatocytes in the liver, cardiac cells in the heart, and neuroglial cells in the brain. This immunocytochemical study also found that MST was localized in both mitochondria and cytoplasm. PMID- 9749959 TI - Endocytosis by the corneal endothelium. I. Regulation of binding and transport of hemeproteins and peroxidase-conjugated lectins across the tissue. AB - Binding, internalization, and movement of hemeproteins and peroxidase-conjugated lectins across organ cultured rat corneal endothelia has been investigated. Horseradish peroxidase (HRP) type II, bound to the surface, was minimally internalized and was easily washed off. In contrast, HRP-VI bound and was rapidly internalized. Reaction product was observed in vesicles, endosomes, multivesicular bodies, and extended along the length of the intercellular space (ICS) to Descemet's membrane. Studies at 4 degrees C indicated HRP-VI bound uniformly along the surface in a punctate fashion. Exposure to polylysine or mannose significantly decreased uptake. Other tracers such as HRP-VIII, -IX, catalase, and microperoxidase exhibited limited uptake by the tissue. However, endothelia vigorously internalized soybean agglutinin (SBA)-HRP, and reaction product was found intracellularly and within the ICS at the cell/Descemet's membrane interface. Internalization and the appearance of SBA-HRP within the ICS was diminished following polylysine or mannose treatment. Experiments at 4 degrees C indicated that SBA-HRP binding and uptake were temperature sensitive. Wheat germ agglutinin (WGA)-HRP was also strongly endocytosed and reaction product was visualized within vesicles, endosomes, and multivesicular bodies. Although WGA-HRP reaction product was observed within the ICS, none was detected at the level of Descemet's membrane. The WGA competitive sugar N-acetyl-D glucosamine, reduced endocytosis, whereas exposure to unlabeled WGA and mannose together reduced uptake. These results indicate endothelia exhibit differential uptake of various hemeproteins and lectins which is dependent on charge, mannose receptors, and appropriate surface sugars. PMID- 9749960 TI - Activation of neurokinin 1 receptors on interstitial cells of Cajal of the guinea pig small intestine by substance P. AB - The aims of this work were to determine whether cells that are similar to the interstitial cells of Cajal (ICC) and have immunoreactivity for the neurokinin 1 (NK1) receptor are indeed ICC; to determine whether the agonist, substance P, binds to and activates the receptor on presumptive ICC; and to investigate the relationship between substance P-immunoreactive nerve fibres and ICC. ICC at the level of the myenteric plexus and in the deep muscular plexus in the duodenum and ileum of the guinea-pig were investigated. Immunoreactivities for the ICC marker, Kit, and the NK1 receptor were colocalised in ICC of the myenteric and deep muscular plexuses. In tissue fixed immediately after its removal from the animal, NK1 receptor-immunoreactive ICC were found at the level of the myenteric plexus in the duodenum, but not in the ileum, and in the deep muscular plexus in the duodenum and ileum. The majority of receptor immunoreactivity was on the cell surface. ICC were exposed to substance P (10(-7) M), initially at 4 degrees C for 1 h to allow the agonist to bind, followed by incubation at 37 degrees C to allow receptor internalisation to proceed. Exposure to substance P caused the NK1 receptor immunoreactivity to aggregate in clumps in the cytoplasm of ICC of the myenteric and deep muscular plexuses, including the ICC of the myenteric plexus of the ileum, where NK1 receptor immunoreactivity was not seen if tissue was not exposed to substance P. Substance P, to which the fluorescent label, cyanine 3.18 (Cy-3), was coupled, bound to the ICC. The Cy-3-substance P was internalised with the receptor following warming to 37 degrees C. Many, but not all, ICC were closely apposed by nerve fibres with immunoreactivity for substance P. It is concluded that the NK1 receptor immunoreactivity on ICC represents receptor that is functional in the sense that it binds the natural agonist substance P and undergoes agonist-induced internalisation. ICC are likely to receive excitatory innervation from the close approaches of tachykinin-containing nerve fibres. PMID- 9749961 TI - Expression of smooth muscle markers in the developing murine lung: potential contractile properties and lineal descent. AB - Contractile cells in the mammalian lung develop in close association with the outgrowing stem bronchi. Fully differentiated smooth muscle cells are typically found in proximal regions, residing in the substantial muscular walls of the major airways and blood vessels. More distally, cells expressing markers of differentiated smooth muscle cells to a variable degree, and which may therefore possess contractile properties, are to be found scattered around the interstitium. We have investigated the temporal and spatial distribution of smooth muscle lineage markers (smooth muscle myosin mRNA) and of those indicative of contractile function (metavinculin mRNA) in the murine lung. In the smooth muscle layers of the bronchi and major blood vessels, these genes are expressed from the onset of pulmonary budding, concurrently with the appearance of alpha smooth muscle actin and calponin proteins. During fetal development, smooth muscle-associated genes and proteins are restricted to this committed smooth muscle population. The first signs of myofibroblast or pericyte differentiation become manifest perinatally, when their expression of alpha-smooth muscle actin escalates. In the adult lung, such cells may be readily pin-pointed by their positive reaction for metavinculin mRNA, but, at maturity, they do not always coexpress alpha-smooth muscle actin. PMID- 9749962 TI - Ultrastructure and cytochemistry of the early calcification site and of its mineralization organic matrix in Paracentrotus lividus (Echinodermata: Echinoidea). AB - The ultrastructure and cytochemistry of skeleton formation sites prior to mineralization are described for the first time in echinoderms. These early sites are intracellular vacuoles located in syncytial pseudopodia of skeleton-forming cells. They contain a mineralization organic matrix, which shows a calcium binding ability and is framed in a tridimensional structure made of concentric layers bridged by radial threads. This organic matrix presents repetitive structures which could be implicated in mineralization control. Both the tridimensional organization of the organic matrix and its framing, before mineralization starts, question the current theories which suggest that the echinoderm organic matrix is soluble at the onset of mineralization and adsorbs on the forming crystal. PMID- 9749963 TI - Secretion of fucosylated oligosaccharides related to the H antigen by human gastric cells. AB - Although the role of the blood group antigens in the gastrointestinal tract is not well understood, alterations in blood group-related antigens have been described in some pathological processes. Thus, the knowledge of their expression under normal conditions is of special interest. Those individuals expressing their ABO blood group in exocrine epithelia and secretions are called secretors. The aim of the present study was the localization of H antigen expression in the normal human gastric epithelial cells of non-O blood group individuals. For this, a monoclonal anti-H antibody was examined by immunocytochemical methods at both the light and electron microscopic levels. In combination with enzymatic and chemical treatments, the nature of the oligosaccharide chains containing the H antigen was characterized. The selected cases were four A secretors, three A nonsecretors, and three B non-secretors. The labeling of the anti-H antibody in the human stomach is described, irrespective of the blood group of the individuals. The staining was abolished when O-linked oligosaccharides were removed. Since commercially available anti-H antibodies usually also recognize other H-related antigens, the labeling of the antibody by H-related antigens cannot be dismissed. Our findings suggest the existence of H or H-related antigens in the O-linked oligosaccharides of the secretory granules of the surface, gastric pit, mucous neck, and transitional cells of the fundic mucosa, and in the intracellular canaliculi and tubulovesicular system of parietal cells. The H or H-related antigens were also localized in the apical membrane of all the cell types of the epithelial cells of the human fundic mucosa. The overall distribution of the H or H-related antigens in the stomach in non-O blood group individuals suggests the constitutive expression of an alpha(1,2)fucosyltransferase. PMID- 9749964 TI - Identification of the taste cell G-protein, alpha-gustducin, in brush cells of the rat pancreatic duct system. AB - The major pancreatic excretory ducts have been shown to contain a large number of specialized epithelial cells, named brush cells, that are characterized by an apical tuft of stiff microvilli. The function of pancreatic brush cells is unknown. Because of some structural similarities to taste receptor cells of the tongue, we addressed the question whether pancreatic brush cells contain the taste cell-specific GTP-binding protein, alpha-gustducin, and hence might be considered to be involved in intraductal chemoreception. By immunostaining, we show that ductal brush cells of the rat pancreatic duct system contain alpha gustducin, which is concentrated in the apical tuft of microvilli and is also found along the basolateral cell surface. A further outcome of this study is that brush cells are concentrated in the terminal portions of extralobular ducts and in the major pancreatic duct where brush cells comprise up to 22% of the ductal epithelium. Immunoblotting of the major pancreatic duct revealed a 42-kDa band that comigrated with alpha-gustducin of the rat tongue. In view of our previous observation that the ductal brush cells are particularly rich in nitric oxide synthase-I, there is reason to assume that these cells might play a role in certain aspects of chemoreceptive signalling. Thus, chemosensory control of pancreatic secretion might occur at two independent sites, the intestine and the terminal portions of the excretory duct system. PMID- 9749965 TI - Haptenylation of antibodies during affinity purification: a novel and convenient procedure to obtain labeled antibodies for quantification and double labeling. AB - Haptenylation of primary antibodies is a useful technique for multiple purposes. It is a technically straightforward procedure, as many haptens are available as N hydroxysuccinimide esters or isothiocyanates. Unfortunately, the hapten group may become covalently attached to or close to the combining site of antibodies, lectins, or other ligand-binding proteins during the process of haptenylation. Thus, the interaction of the corresponding protein with its ligand may become severely hampered. To overcome this restriction, we developed a novel procedure for the haptenylation of polyclonal antibodies that combines purification and haptenylation. Haptenylation during adsorption to the affinity matrix combines two advantages: the antigen binding site is protected and the labeling procedure becomes most convenient, as overlabeled proteins and unreacted haptens are easily removed by simple washing. Haptenylation during adsorption to the affinity matrix is a two-phase reaction, which requires different conditions to the conventional procedure. To obtain such optimal conditions, stabilities and reactivities of N hydroxysuccinimide esters and isothiocyanate groups were investigated with a newly developed assay. Based on this information, antibodies against two recently described calcium-binding proteins, NCS-1 and NVP-3, were biotinylated or digoxigenylated. The haptenylated antibodies were successfully applied for biochemical determination and simultaneous immunoenzymatic double labeling of the two proteins. PMID- 9749967 TI - Occupational health in Singapore. AB - Singapore, a newly industrializing country in Southeast Asia, has a resident population of 3 million and a work force of 1.75 million. Most workers are employed in the manufacturing, services, and commerce sectors. Agricultural and mining activities are negligible. In 1996 the infant mortality rate was 3.8 per 1,000 live births and the life expectancy at birth was 77 years. In 1996 the total industrial accident rate was 2.7 per million man-hours worked and the severity rate was 353 industrial man-days lost per million man-hours worked. The shipbuilding and construction industries had the most frequent and most severe accidents. In the same year, 1,521 cases of occupational disease were notified to, and confirmed by, the Ministry of Labor. The majority of cases involved noise induced hearing loss. There is substantial underreporting of cases. New cases that are expected to appear will be work-related illnesses such as musculoskeletal or psychosocial disorders. The principal occupational health legislation in Singapore is the Factories Act. Although it selectively targets workers at highest risk of developing occupational illness, its main limitation is the exclusion of nonfactory workers, who comprise 63% of the working population. Labor regulations are enforced by the Ministry of Labor. Workmen's compensation paid in 1995 amounted to S $46.6 million (U.S. $1=S $1.75). Education and training in occupational health is provided by employer federations, employee unions, and various government agencies. Occupational health is taught to medical students during their undergraduate training. Postgraduate-diploma and Masters programs in occupational medicine are also available. About 600 doctors in Singapore have some form of postgraduate training in occupational health. Health care for workers is offered either through the private sector or through government clinics and hospitals. Although Singapore has made great strides in protecting and promoting the health of its workers, it must constantly strive to strengthen its commitment to occupational health and safety. New problems in the next century must be anticipated and solutions, implemented. Improved training and development of health professionals is needed such that they be better prepared to deliver optimal occupational health care. Finally, labor legislation must be appropriate and responsive to protect the health of all workers. PMID- 9749966 TI - A new method for flat-embedding large native cryostat sections for targeting small preneoplastic lesions in comparative ultrastructural and ultracytochemical investigations. AB - Ultrastructural studies of rare and small cellular lesions in pathologically altered tissue are difficult to perform by applying conventional electron microscopic preparation. The search for lesions, often consisting of only a few cells in randomly obtained small specimen blocks, is time consuming and often without success. The methodological requirements for comparative enzyme cytochemical and morphological studies, i.e., preservation of both enzyme activity and ultrastructure, are divergent. By processing large native cryostat sections for electron microscopy, small preneoplastic focal lesions were successfully targeted in liver and kidney. Glucose-6-phosphatase, alkaline phosphatase, acid phosphatase, catalase, and cytochrome c oxidase activities were distinctly localized to endoplasmic reticulum, canalicular membrane, lysosomes, peroxisomes, and mitochondria, respectively, in the morphologically altered cells. Fixation of serial cryostat sections and enzyme reactions were both carried out through a semipermeable membrane except those for cytochrome c oxidase, which was demonstrated after fixation through the membrane by floating the section in incubation medium containing cytochrome c. Thereafter, the sections were flat embedded and polymerized between epoxy resin disks and aluminum dishes fitting exactly together. The objects of interest were identified in the light microscope, cut out, and reembedded in reversed gelatine capsules. By using this technique an ultrastructural preservation was achieved similar to that seen after immersion fixation. The enzyme activities were clearly localized without diffusion of the reaction product or unspecific deposits. The procedure permits precise targeting and complex studies of rare and small lesions, and opens new perspectives for the use of cryo-preserved tissue. PMID- 9749968 TI - High-pressure liquid chromatographic determination of toluene in urine as a marker of occupational exposure to toluene. AB - OBJECTIVE: To establish a convenient method by high-pressure liquid chromatography (HPLC) to measure toluene in urine as a marker of occupational exposure to toluene. METHODS: As soon after sampling as possible, 1 ml of urine was mixed with an equal volume of acetonitrile in a 2.2-ml HPLC glass bottle, and the bottle was tightly sealed and stored at 4 degrees C. Immediately before HPLC determination, 100 microl methanol was added to the mixture to prevent confounding effects of glycosuria, and the bottle was spun to remove any suspended matter. An aliquot of the supernate was introduced into the HPLC system and analyzed on a PRODIGY column, with an acetonitrile - perchloric acid phosphoric acid - water mixture serving as the mobile phase. The effluent was monitored at 191 nm. RESULTS: The method can measure toluene in urine every 20 min, the detection limit was 2 microg/l, the coefficient of variation was less than 5%, and the recovery rate was 100%. No significant reduction in toluene concentration was observed for 1 week after storage at 4 degrees C. When the method was applied to end-of-shift urine samples from 13 male workers exposed to toluene at 18-140 ppm and also to urine samples from 10 nonexposed male controls, toluene in urine was linearly related to toluene exposure concentration, with a regression line passing close to the origin. The correlation coefficient was as high as 0.97 (n=23). No toluene was detected in control urine samples. Calculations suggest that urinary toluene accounts for as little as less than 0.01% of the toluene absorbed via inhalation and that the absorbed toluene is converted almost quantitatively to hippuric acid and, by less than 0.1%, to o cresol. PMID- 9749969 TI - N,N-dimethylformamide--influence of working conditions and skin penetration on the internal exposure of workers in synthetic textile production. AB - OBJECTIVES: This study examined the external and internal exposure to the solvent N,N-dimethylformamide (DMF) of 126 workers from a factory producing synthetic fibers. METHODS: Air measurements were carried out using personal air samplers with diffusion tubes (Drager, ORSA 5). For the purpose of biological monitoring the levels of N-methylformamide (NMF) in urine were measured in preshift and postshift samples. Determinations were carried out using gas chromatography. Anamnestic data were collected with standardized questionnaires, including personal data, working history and current working conditions, and former and current illness with regard to the effects of DMF. Skin diseases were documented by a dermatologist. RESULTS: DMF concentrations measured in the air ranged between <0.1 and 37.9 ppm (median 1.2 ppm). Concentrations of NMF varied from 0.05 to 22.0 mg/l (preshift values) and from 0.9 to 100.0 mg/l (postshift values). The creatinine-related values (0.02-44.6 mg/g preshift; 0.4-62.3 postshift) were subject to less variation and therefore represented the level of exposure better than the values related to volume. Additional investigation of a subcollective (n=31) over a period of 4 days showed that NMF did not accumulate in the organism. The positive but relatively weak association observed between the DMF concentrations measured in the workplace air and the values recorded for internal exposure in this study can be explained by influencing factors such as dermal absorption or protective clothing. Interindividual differences in internal exposure were found for the specific work areas. The German BAT value (15 mg NMF/l urine) was exceeded in 36 persons (29%) despite the use of breathing protection and protective gloves, without increased values being measured in the air. Increased absorption without higher-level exposure could particularly also be observed in employees with eczema. CONCLUSIONS: From the point of view of the prevention of disease, biological monitoring is the best instrument for exposure assessment of workers exposed to DMF. PMID- 9749970 TI - Increased acquired dyschromatopsia among solvent-exposed workers: an epidemiology study on 249 employees of an aluminum-foil printing factory. AB - OBJECTIVE: To analyze the effects on color vision of chronic exposure to mixtures of solvents including ethyl acetate, ethanol, and ketones among the workers of a large factory specializing in the manufacture of photoengraved aluminum packaging. METHODS: We analyzed a group of 129 subjects who had been exposed to solvents for more than 3 years (mean age 40 years, range 25 59 years) and a nonexposed group of 120 subjects (mean age 41 years, range 21-57 years). The two groups had a similar length of service (17 and 19 years on average, respectively). The exposed subjects consumed more tobacco and alcohol than the nonexposed workers. The study involved administration of the Lanthony D-15 desaturated test. The subjects were classified as having or not having dyschromatopsia on the basis of Lanthony's criteria and the chromatic confusion index (CCI) was calculated according to Bowman's method. After observation of the work stations and analysis of an occupational questionnaire the findings were used to construct a cumulative exposure index covering the whole of each subject's working life. Air samples were taken at each work station and in each production sector to determine current exposure. RESULTS: The average cumulative exposure index was 194 (median 174, range 27-513). The average hygienic effect index (according to ACGIH recommendations), regularly used for atmospheres containing mixtures of products, varied between 12% and 27% of the occupational limit values permitted under French legislation. As regards color vision, 23% of the exposed group had dyschromatopsia, as did 13% of the nonexposed group, with the odds ratio (OR) adjusted for age and consumption of tobacco and alcohol being 1.99 (1.02, 3.89). The analysis of the dose-response relationship according to the cumulative exposure index showed an OR of 1.59 for the lowly exposed group (index < 174) and an OR of 2.40 for the highly exposed group (index > 174) as compared with the nonexposed group. Subjects with complex color vision loss (n=23) had a significantly higher cumulative exposure index than those with blue yellow loss (n=23; 232 versus 155; P=0.04). The CCI was on average higher in the exposed group, but the difference between the two groups was not significant. CONCLUSION: The results of this study are in favor of an increased risk for impairment of chromatic discrimination among workers exposed to mixtures of solvents including mainly ethyl acetate, ethanol, and ketones. PMID- 9749971 TI - Breath, urine, and blood measurements as biological exposure indices of short term inhalation exposure to methanol. AB - Due to their transient nature, short-term exposures can be difficult to detect and quantify using conventional monitoring techniques. Biological monitoring may be capable of registering such exposures and may also be used to estimate important toxicological parameters. This paper investigates relationships between methanol concentrations in the blood, urine, and breath of volunteers exposed to methanol vapor at 800 ppm for periods of 0.5, 1, 2, and 8 h. The results indicate factors that must be considered for interpretation of the results of biological monitoring. For methanol, concentrations are not proportional to the exposure duration due to metabolic and other elimination processes that occur concurrently with the exposure. First-order clearance models can be used with blood, breath, or urine concentrations to estimate exposures if the time that has elapsed since the exposure and the model parameters are known. The 0.5 to 2-h periods of exposure were used to estimate the half-life of methanol. Blood data gave a half life of 1.44+/-0.33 h. Comparable but slightly more variable results were obtained using urine data corrected for voiding time (1.55+/-0.67h) and breath data corrected for mucous membrane desorption (1.40+/-0.38 h). Methanol concentrations in blood lagged some 15-30 min behind the termination of exposure, and concentrations in urine were further delayed. Although breath sampling may be convenient, breath concentrations reflect end-expired or alveolar air only if subjects are in a methanol-free environment for 30 min or more after the exposure. At earlier times, breath concentrations included contributions from airway desorption or diffusion processes. As based on multicompartmental models, the desorption processes have half-lives ranging between 0.6 and 5 min. Preliminary estimates of the mucous membrane reservoir indicate contributions of under 10% for a 0.5-h exposure and smaller effects for longer periods of exposure. PMID- 9749972 TI - The effects of an ergonomic-educational course. Postural load, perceived physical exertion, and biomechanical errors in nursing. AB - OBJECTIVES: To evaluate the results of an ergonomic-educational course for nurses we assessed the number and percentage of harmful postures and of ergonomic and biomechanical errors made before and after the course. We also studied the perceived physical exertion. MEANS AND METHODS: In all, 12 nurses who had participated in the course (trainees) and 12 who had not (controls) were recorded on video while performing standardized nursing tasks. The wards from which the two groups of nurses came were comparable, as were the patient populations. The nurses were also comparable in some personal characteristics. The tasks they performed included washing, lifting, and repositioning a patient as well as certain tasks other than patient handling. Video recordings were made once before (1-2 weeks) and twice after the course (after 3 months and after 15 months). The harmful postures, the errors made, and the ratings of perceived exertion were measured by means of the Ovako Working-posture Analysis System (OWAS), a checklist, and Borg scores, respectively. RESULTS: When the first and last measurements of all the above tasks taken as a whole were analyzed the trainees showed a significant improvement in the number and percentage of harmful postures and errors, whereas the controls did not. The same could be concluded for lifting alone. After the course the new work routine did not appear to have caused any extra perceived physical exertion. CONCLUSION: It can be concluded that the course was successful, although it should be carefully investigated as to whether nurses remain capable of working safely in daily practice. The work pressure that nurses experience during their normal duties could prevent them from working safely during everyday work. PMID- 9749973 TI - Exposure to polymeric materials in vascular soft-tissue sarcomas. AB - Known etiologic factors related to endothelial angiosarcomas are exposures to arsenic, thorium dioxide, therapeutic irradiation, and certain congenital diseases. Little is known on the etiology of hemangiopericytomas. Since 1974, several reports have appeared on a distinct relationship between the exposure to vinyl chloride monomers and angiosarcomas of the liver. The early reports on this matter provided the reason to collect the occupational histories of vascular sarcomas accumulated since that time. METHODS: Data on the occupational histories of patients with different forms of angiosarcomas, treated between 1975 and 1995 in two institutions, were prospectively collected and analyzed. In this personal series the only selection criteria were the referral of patients for postoperative or palliative irradiation and their personal care by the author. FINDINGS: Among 21 adult cases of vascular sarcomas there were 4 patients with occupational exposure to vinyl chloride (VC) either alone or together with other artificial polymers. Seven other patients showed exposure to several plastics or resins other than VC. Altogether, 11 of 21 (52%) of the explored patients were found to have been exposed to artificial polymeric materials over a mean period of 18 years. The patients without such exposure were 4 farmers, 2 house-wives, and 1 woodworker, telephonist, mason, and inland revenue official, respectively. Two cases were radiation-induced. The series contained no angiosarcoma of the liver. INTERPRETATION: This study offers new evidence of the occurrence of vinyl chloride-induced angiosarcomas outside the liver and confirms observations that have previously been published in case reports. Moreover, it may be suspected from this analysis that polyvinyl chloride and its monomers are not the only polymeric materials that may contribute to an induction of angiosarcomas in humans. Repeated occupational histories have to be taken from the patients to achieve data of the greatest value, since there are many professional activities that do not primarily lead to the assumption of specific exposure to polymeric materials. PMID- 9749974 TI - Urinary N-acetyl-beta-glucosaminidase as an indicator of renal dysfunction in electroplating workers. AB - OBJECTIVES: To investigate chromium-induced renal dysfunction in electroplating workers. METHODS: A cross-sectional study was used to evaluate four biochemical markers of renal function. A total of 178 workers were divided into 3 comparable groups consisting of 34 hard-chrome plating workers, 98 nickel-chrome electroplating workers. and 46 aluminum anode-oxidation workers, who represented the reference group. Ambient and biological monitoring of urinary chromium were performed to measure exposure concentrations. RESULTS: Overall, urinary chromium concentrations were highest among hard-chrome plating workers (geometric mean 2.44 microg/g creatinine), followed by nickel-chrome electroplating workers (0.31 microg/g creatinine) and aluminum workers (0.09 microg/g creatinine). Airborne chromium concentrations were also highest in the hard-chrome plating area (geometric mean 4.20 microg/m3), followed by the nickel-chrome electroplating area (0.58 microg/m3) and the aluminum area (0.43 microg/m3). A positive correlation was found between urinary chromium and airborne concentrations (r=0.54, P < 0.01). Urinary concentrations of N-acetyl-beta-D-glucosaminidase (NAG) were also highest among hard-chrome plating workers (geometric mean 4.9 IU/g creatinine), followed by nickel-chrome workers (3.4 IU/g creatinine) and aluminum workers (2.9 IU/g creatinine). The prevalence of "elevated" NAG (>7 IU/g creatinine) was significantly highest among hard-chrome plating workers (23.5%), then among nickel-chrome workers (7.1%) and aluminum workers (8.7%). Differences in beta2-microglobulin, total protein, and microalbumin were not significant. CONCLUSION: The author's evidence indicates that NAG is an early indicator of renal dysfunction in hard-chrome plating workers. PMID- 9749975 TI - Sick-building symptoms in office workers in northeastern France: a pilot study. AB - STUDY OBJECTIVES: To verify that sick building symptoms are present in north eastern France office workers; to try to identify new confounding factors. METHODS: The design was that of a cross-sectional study with control group. We studied with the same methods the personnel of an air-conditioned building (n=425), and of a naturally ventilated building (n=351). Air temperature and humidity, bacterial and fungal densities were measured by the same technical staff in the two buildings. A standard questionnaire on irritative and respiratory symptoms, personal and family history, and lifestyle was completed by the participants. RESULTS: In univariate analysis, exposure to air-conditioning was associated with an increased prevalence of symptoms (odds ratios-OR-between 1.54 and 2.84). A significant increase in sickness absence was also found among subjects working in air-conditioned offices. As a series of factors were suspected to interfere with these associations, logistic regression was applied. This method confirmed exposure to be an independent determinant of 7 symptoms, and also identified two determinants not previously described: a family history of respiratory diseases and "do-it-yourself' activities. IN CONCLUSION: we found the sick building symptoms to be present in a group of French office workers exposed to air-conditioning. We confirmed the influence of a number of confounding factors and described two further confounders - do-it-yourself activities at home and a history of familial respiratory disease. PMID- 9749976 TI - Visual function in professional truck drivers. AB - OBJECTIVES: The purpose of this study was to examine the visual function of professional truck drivers at working age to find out whether older drivers had any defective function and should therefore be given less demanding duties at work. METHODS: Of the 100 drivers invited to the study, 77 came to the examination, including 74 men and 3 women aged from 30 to 66 years (mean 50.3+/ 10.3 years). In addition to the basic eye examination, visual fields, dark adaptation, contrast sensitivity, color vision, and glare sensitivity were studied. RESULTS: Two drivers (2.6%) had an incipient cataract in one eye, four (5.2%) had slight fundus abnormalities, five (6.5%) had exo- or esotropia, and five (6.5%) had amblyopia. Visual acuity in the better eye varied from 0.4 to 1.2 (mean 1.06+/-0.17) and in the other eye from 0.1 to 1.2 (mean 0.96+/-0.23). Five drivers (6.5%) had inadequate visual acuity for a professional driver's license. Visual fields were interpreted as normal in all drivers. The results of the dark adaptation, contrast-sensitivity, and glare testing showed values within normal ranges for all drivers. In the color-vision tests, five male drivers (6.8% of the men) had a slight congenital green defect, and two drivers had an acquired blue defect in one eye because of cataract and diabetes. CONCLUSIONS: According to the present study, there is no need to give older drivers less demanding duties because of their eyes. However, one serious finding in the present study needs attention: the visual acuity was lower than that required for a professional driver's license in five drivers. Evidently, more regular checkups of visual acuity are needed for a professional driver's license. PMID- 9749977 TI - Importance of receptor binding in the immunogenicity, adjuvanticity and therapeutic properties of cholera toxin and Escherichia coli heat-labile enterotoxin. PMID- 9749978 TI - Balance of proinflammatory and antiinflammatory cytokines in mice immunized with Escherichia coli and correlation with mortality after lethal challenge. AB - The balance of proinflammatory and antiinflammatory cytokines, their correlation with endotoxin levels and mortality rate after lethal challenge of Escherichia coli was investigated in mice immunized weekly for 8 weeks with formalin-killed E. coli either untreated or treated with 0.5x minimal inhibitory concentration of aztreonam. Control mice treated in parallel with saline, died within 24 h after challenge with 100x lethal dose (LD50) of viable E. coli O6:K-. Mice immunized with antibiotic-treated bacteria showed a significantly higher survival than mice immunized with untreated E. coli. Cytokines were not detected in the sera of control mice during the entire period of immunization. At 90 min after immunization, mice immunized with antibiotic-treated E. coli showed tumor necrosis factor-alpha (TNF-alpha) levels significantly lower and interleukin (IL) 6 levels significantly higher (P < 0.05) than mice immunized with untreated E. coli, while comparable levels of interferon-gamma (IFN-gamma were measured in both groups. TNF-alpha and IL-10 levels measured 90 min after lethal challenge correlated with the mortality rate observed in each group (r = 0.96 for TNF-alpha and 0.94 for IL-10). IL-6 levels correlated with survival (r = 0.95), while IFN gamma serum levels did not differ in the two immunized groups, but were significantly higher than those measured in the control mice. IL-4 was detected only after challenge of mice immunized with antibiotic-treated bacteria. Comparable levels of circulating endotoxin were measured after lethal challenge in both control and immunized mice. These data showed that in the presence of a protective immune response the survival of immunized mice was correlated with an early alteration of cytokine expression pattern including enhanced secretion of IL-4, IL-6 and IFN-gamma, and reduced secretion of TNF-alpha and IL-10. PMID- 9749979 TI - Proliferation and MHC-unrestricted bystander lysis by virus-specific cytotoxic T cells following antigen self-presentation. AB - Cytotoxic T cells (CTL) not only act as effector cells, but can also serve as antigen-presenting cells (APC) for other CTL due to their expression of major histocompatibility complex (MHC) class I molecules. In the present study we show that independently derived CTL lines (CTLL) with specificity for an L(d) presented nonapeptide corresponding to amino acids 168-176 of the immediate-early 1 (IE1) protein of murine cytomegalovirus not only lyse syngeneic but also allogeneic target cells, if the peptide is present during the cytolytic assay. Whereas a short peptide pulse is sufficient to render syngeneic cells susceptible to lysis, continued presence of soluble peptide is mandatory for the lysis of allogeneic target cells. This indicates a difference in the mechanisms involved. Syngeneic BALB/c B cell blasts (K(d)D(d)L(d)) and mutant BALB/c-H-2dm2 B cell blasts lacking the restricting Ld molecules (K(d)D(d)0) were lysed to a similar extent in the absence of the IE1 nonapeptide, provided that the IE1-specific CTL had been pre-incubated with the peptide before the cytolytic assay. Since the mutant cells cannot present the IE1 peptide, their lysis indicates an MHC unrestricted, peptide-independent mode of recognition by the CTLL. In addition, proliferation of the CTLL takes place after incubation with the cognate peptide, even in the absence of professional APC. These data indicate inter-CTL antigen self-presentation, resulting in activation of the lytic machinery leading to peptide-independent bystander lysis of allogeneic as well as syngeneic target cells. PMID- 9749980 TI - Identification and localization of Chlamydia pneumoniae in the Alzheimer's brain. AB - We assessed whether the intracellular bacterium Chlamydia pneumoniae was present in post-mortem brain samples from patients with and without late-onset Alzheimer's disease (AD), since some indirect evidence seems to suggest that infection with the organism might be associated with the disease. Nucleic acids prepared from those samples were screened by polymerase chain reaction (PCR) assay for DNA sequences from the bacterium, and such analyses showed that brain areas with typical AD-related neuropathology were positive for the organism in 17/19 AD patients. Similar analyses of identical brain areas of 18/19 control patients were PCR-negative. Electron- and immunoelectron-microscopic studies of tissues from affected AD brain regions identified chlamydial elementary and reticulate bodies, but similar examinations of non-AD brains were negative for the bacterium. Culture studies of a subset of affected AD brain tissues for C. pneumoniae were strongly positive, while identically performed analyses of non-AD brain tissues were negative. Reverse transcription (RT)-PCR assays using RNA from affected areas of AD brains confirmed that transcripts from two important C. pneumoniae genes were present in those samples but not in controls. Immunohistochemical examination of AD brains, but not those of controls, identified C. pneumoniae within pericytes, microglia, and astroglia. Further immunolabelling studies confirmed the organisms' intracellular presence primarily in areas of neuropathology in the AD brain. Thus, C. pneumoniae is present, viable, and transcriptionally active in areas of neuropathology in the AD brain, possibly suggesting that infection with the organism is a risk factor for late onset AD. PMID- 9749981 TI - Role of curdlan sulfate in the production of beta-chemokines and interleukin-16. AB - The blocking effect of curdlan sulfate (CRDS) on human immunodeficiency virus (HIV) infection has been thought to be related to inhibition of the binding of HIV-1 envelope glycoprotein (gp120) and CD4 molecules. However, recent reports have indicated that blocking the binding of gp120 to CD4 by CRDS only makes a small contribution to the inhibition of HIV-1 infection. We report here that the effect of CRDS on the production of beta-chemokines and cytokines might be important in the inhibition of HIV-1 infection, in addition to interference with the binding of gp120 to CD4+ cells. PMID- 9749982 TI - Expression of CTLA-4 (CD152) on human medullary CD4+ thymocytes. AB - CTLA-4 (CD152) is a T cell surface receptor with sequence homology to the co stimulatory molecule CD28. The molecule, which is essential for the inhibitory regulation of the immune response, becomes transiently expressed on mature T cells after stimulation in vitro. In situ, CTLA-4+ T cells are enriched in the light zones of the germinal centers in human peripheral lymphoid organs. In this study we have studied expression of CTLA-4 in human thymus in situ. CTLA-4 was expressed on about one third of CD4+/CD8-/CD1- medullary thymocytes. CTLA-4 was acquired by a subset of immature (CD1+) thymocytes and lost from the mature (CD1 ) subpopulation within 48 h of cell culture, suggesting that the expression on medullary thymocytes is transient. The demonstration of CTLA-4 on a substantial subpopulation of mature CD4+ thymocytes adds a new dimension to the understanding of this important molecule. When contemplating application of anti-CTLA-4 for therapy its potential influence on T cell maturation has to be taken into account. PMID- 9749983 TI - Wortmannin blocks Yersinia invasin-triggered internalization, but not interleukin 8 production by epithelial cells. AB - In response to bacterial infection epithelial cells up-regulate expression and secretion of proinflammatory cytokines. Previous work from our laboratory showed that basolateral infection of polarized T84 cells with Yersinia enterocolitica induces interleukin-8 (IL-8) secretion in the absence of significant invasion. Here we studied Y. enterocolitica-induced IL-8 secretion by epithelial HeLa cells as a function of Yersinia invasion or adhesion. For this purpose we tried to separated induction of IL-8 secretion from invasion by treating HeLa cells with signal transduction inhibitors prior to infection. While staurosporin and genistein inhibited both Yersinia invasion and Yersinia-triggered IL-8 secretion, wortmannin, an inhibitor of the phosphatidylinositol-3-phosphate kinase (PI3-K), blocked invasion of Y. enterocolitica into HeLa cells but did not show any effect on IL-8 secretion. These results suggest that Yersinia adhesion might be sufficient to induce IL-8 secretion by epithelial cells. Further analysis demonstrated the requirement of the Yersinia invasion locus inv for adhesion mediated induction of IL-8 secretion. Thus, HeLa cells infected with an E. coli strain expressing the Y. enterocolitica inv locus induced IL-8 secretion in the presence and absence of wortmannin. Reverse transcription-polymerase chain reaction analysis revealed that adhesion of inv-expressing Y. enterocolitica or E. coli results in the transcriptional activation of the IL-8 gene. These results suggest that Y. enterocolitica adhesion to host cells via Inv activates de novo synthesis and secretion of IL-8. PMID- 9749984 TI - Attentional selection of motion states. AB - In the present experiments the ability of attention to modulate the perceptual dominance in ambiguous motion displays was assessed, as well as the resulting changes of the motion aftereffect (MAE). As ambiguous motion stimuli we used plaids consisting of two differently moving components, where directed attention can select one or the other of the two components. We monitored this selection process during adaptation, and then measured the resulting MAE with static and/or dynamic test stimuli. The results demonstrate that attention increased the dominance of the selected component substantially. This led to an increase in the size of the MAE to the attended component, while the MAE to the not attended component decreased. It was further shown that the attentional MAE effect was relatively short-lasting, only for about the first third of the total MAE. It is concluded that attention can access levels of motion processing that are prior to or at the level of the generation of the component MAEs. PMID- 9749985 TI - The effects of eccentricity and spatial frequency on the orientation discrimination asymmetry. AB - This study investigated the effects of eccentricity and spatial frequency on the discrimination of vertical and oblique (10 deg from vertical) Gabor patches. Within a display stimuli were scaled by a factor F = 1 + E/E2 at each eccentricity (E) in an attempt to equate either the number of photoreceptors (E2 = 2.5) or cortical area (E2 = 0.77) engaged at each eccentricity. The task was to detect a differently oriented target among eleven distractors. Orientation discrimination asymmetries (ODAs) were found such that an oblique stimulus was easier to detect in a background of vertical stimuli than vice versa. Subjects were equally sensitive to the two highest frequency Gabor patches and less sensitive to the lowest frequency Gabors. When stimuli were scaled with E2 = 2.5 sensitivity was constant at all eccentricities and the ODA magnitude was unaffected. When stimuli were magnified with E2 = 0.77 both sensitivity and ODA magnitude increased with eccentricity. Generally, we may conclude that the ODA effect is not a strictly foveal phenomenon nor is it a strictly high frequency effect. PMID- 9749986 TI - The effects of attentional spread and attentional effort on orientation discrimination. AB - The spatial properties of visual attention and its relation to attentional effort have been investigated, and it has been found that stimulus detectability changes as a function of attentional beam width and degree of task difficulty. Using a matching-to-sample paradigm, two Gabor patches were presented simultaneously both as a sample stimulus and a test stimulus, the stimuli set at three different distances. Task difficulty was gradated by changing the orientation difference of the two Gabor patches on nonmatching trials. 'Difficult' nonmatching probe trials were embedded within an easy block of trials (easy condition), and vice versa for 'easy' probe trials. The detectability, d', differences of probe trials in the two conditions were calculated as a measure of change in attention. Our results show that the detectability of a pair of stimuli decreases with an increase in the distance between stimuli. Furthermore, the results indicate an increase in attentional effort for various attentional beam widths and that r = d'easy/d'difficult of the probe trials is constant throughout the different stimuli separation. PMID- 9749987 TI - On the perception of objects and their orientations. AB - A novel paradigm, description-depiction classification (DDC), was used to test whether knowledge of object orientation in the picture plane precedes or follows object identification. Undergraduate students were asked to verify the identities and orientations of depicted objects against preceding descriptions, e.g. 'UPRIGHT CAR'. The results showed that identity mismatches were verified faster and more accurately than orientation mismatches, regardless of whether subjects had to discriminate small (90 deg) or large (180 deg) differences in orientation. These findings suggest there is primacy for identity information in that (a) subjects determine object identity before they determine object orientation, and (b) even when orientation is determined, identity information tends to dominate the response. This is the signature of a perceptual-cognitive system that has evolved to rapidly identify objects, not their orientations, contra to theories of recognition that assume orientation is determined before an object has been classified at the basic level. PMID- 9749988 TI - The warped geometry of visual space near a line assessed using a hyperacuity displacement task. AB - Badcock and Westheimer (Spatial Vision 1 (1), 3-11, 1985) showed that a thin vertical line induces nearby zones of attraction and repulsion; this study extends those results by more closely examining the horizontal and vertical extents of the repulsion zone and by using an illusory contour to induce repulsion. The experimental paradigm measures perceived hyperacute displacements of a thin vertical line 10' tall. Halfway through the stimulus, the bright target line was shifted and a lower contrast flanking line added. Conditions equivalent to Badcock and Westheimer replicate their results. Repulsion is observed horizontally from separations of 5' to at least 30' and becomes minimal at 50'. Repulsion also decreases with increasing vertical separation. Another experiment shows that symmetry is not required for repulsion when the flanking line is split into two vertically separated fragments: one fragment alone causes the same amount of repulsion as both fragments together. Finally, it is shown that a flanking contour formed by the grating illusion causes repulsion of the target line in the same manner as a target line defined by luminance. PMID- 9749989 TI - Mast cells of psoriatic and atopic dermatitis skin are positive for TNF-alpha and their degranulation is associated with expression of ICAM-1 in the epidermis. AB - The release of cytokines from cutaneous cells may be of major importance in the initiation and development of many inflammatory skin disorders. For example, tumor necrosis factor-alpha (TNF-alpha), which in healthy skin is found preformed only in mast cells, is able to induce the expression of several adhesion molecules including intercellular adhesion molecule-1 (ICAM-1). Increased expression of ICAM-1 occurs in keratinocytes in lesional skin of psoriasis and atopic dermatitis (AD) and it is considered to be an important initiator of leucocyte/keratinocyte interactions in skin inflammation. We counted the mast cells showing TNF-alpha immunoreactivity using a double-staining method in nonlesional and lesional skin sections from 12 patients with AD and 12 patients with psoriasis. The percentage of TNF-alpha+ mast cells in lesional and nonlesional AD skin was 36 +/- 22% and 21 +/- 15% (P < 0.018, paired t-test), respectively, and in psoriatic skin was 16 +/- 25% and 15 +/- 15%, respectively (P < 0.89, paired t-test). We also cultured whole skin biopsies taken from the healthy-looking skin of psoriatic and AD patients in the presence of mast cell degranulator compound 48/80, which resulted in focal expression of ICAM-1 in the epidermis. In cultured keratinocytes, both histamine and an extract of a human mast-cell line (HMC-1) induced ICAM-1 immunostaining only in occasional cells, but the combination of histamine and the HMC-1 extract resulted in intense ICAM-1 staining in numerous cells. This enhancement of ICAM-1 staining was abolished by preincubation of the HMC-1 extract with anti-TNF-alpha antibody. These results suggest that the degranulation of mast cells induces the expression of ICAM-1 in keratinocytes probably via TNF-alpha and histamine. PMID- 9749990 TI - The monoclonal antibody AS02 recognizes a protein on human fibroblasts being highly homologous to Thy-1. AB - Recently, we described a novel fibroblast-restricted monoclonal antibody (mAb AS02) that recognizes a membrane-bound antigen. Characterization and isolation of the corresponding antigen showed that mAb AS02 recognized a protein on human fibroblasts that is highly homologous or identical to human Thy-1 antigen (CD90). Partial amino acid sequencing of the corresponding mAb AS02 antigen and comparison with known proteins revealed a 100% homology of the sequenced peptides to the human Thy-1 antigen. Cross-immunodepletion studies with mAb AS02 and an anti-Thy-1 antibody confirmed these results. Utilizing two-dimensional (2D) gel electrophoresis of fibroblast cell extracts and purified antigen, mAb AS02 and the anti-Thy-1-antibody recognized identical protein spots. Furthermore, we demonstrated many identical biochemical properties of the corresponding AS02 antigen and Thy-1 antigen, such as the molecular weight of the core protein and deglycosylation products and the detection of a GPI anchor. In functional assays, the attachment of fibroblasts to collagen I and fibronectin was increased after incubation of fibroblasts with mAb AS02. Therefore, the Thy-1 antigen appears to be involved in the regulation of the adherence of human dermal fibroblasts. PMID- 9749991 TI - IL-1 and IL-1 receptor antagonist regulation during keratinocyte cell cycle and differentiation in normal and psoriatic epidermis. AB - Changes in the levels of IL-1 (IL-1alpha, IL-1beta, and its receptor antagonist, IL-1RA) occur upon keratinocyte differentiation in vitro and are associated in vivo with abnormal differentiated and hyperproliferative states of psoriatic keratinocytes. A flow cytometric procedure, capable of detecting changes in the intracellular levels of IL-1, was used to determine whether intracellular IL-1/IL 1RA levels in psoriatic and normal keratinocytes alter during in vivo differentiation and the cell cycle. Increases in the IL-1RA levels and IL-1alpha levels were observed as both normal and psoriatic keratinocytes differentiated from basal stem cells (beta1 integrin+, small size) into transient amplifying cells (TAC; beta1 integrin+, large size). Upon further differentiation (beta1 integrin-, large size) both IL-1RA and IL-1alpha levels dropped. However, while psoriatic IL-1beta levels increased as cells differentiated into TACs, little change occurred in the IL-1beta levels of normal keratinocytes during differentiation. Changes in IL-1/IL-1RA levels were also detected as keratinocytes progressed through the cell cycle. Within the basal stem cell population of both normal and psoriatic keratinocytes, the IL-1alpha and IL-1RA levels increased between G0/G1 and S but not between S and G2/M. However, psoriatic basal keratinocyte IL-1beta levels differed from those of normal keratinocytes by showing no increase between S and G2/M. The IL-1/IL-1RA levels of normal TAC increased throughout the cell cycle. However, in psoriatic TAC, a slight decrease in IL-1alpha and IL-1RA levels was observed between G0/G1 and S followed by a delayed increase between S and G2/M. IL-1beta levels in psoriatic TAC varied little throughout the cell cycle. Thus, we were able to detect precisely the regulation of IL-1/IL-1RA intracellular levels during the keratinocyte cell cycle and differentiation, showing notably decreased IL-1beta upregulation in psoriatic keratinocytes progressing through the cell cycle. PMID- 9749992 TI - Linoleic acid and alpha-linolenic acid lightens ultraviolet-induced hyperpigmentation of the skin. AB - This study was conducted to evaluate the effects of unsaturated fatty acids on ultraviolet-induced hyperpigmentation of the skin. An efficient lightening effect was observed following topical application of linoleic acid or alpha-linolenic acid to UV-stimulated hyperpigmented dorsal skin of brownish guinea pigs. The number of melanocytes in the treated skin was similar to the number in the skin of the pigmented control, indicating that the pigment-lightening effect was not due to depletion of melanocytes. In vitro experiments using cultured murine melanoma cells showed that melanin production was inhibited most effectively by alpha-linolenic acid, followed by linoleic acid and then by oleic acid. Furthermore, the turnover of the stratum corneum, which plays an important role in the removal of melanin pigment from the epidermis, was accelerated by linoleic acid and by alpha-linolenic acid. Taken together, the results suggest that the pigment-lightening effects of linoleic acid and alpha-linolenic acid are, at least in part, due to suppression of melanin production by active melanocytes, and to enhanced desquamation of melanin pigment from the epidermis. PMID- 9749993 TI - The sodium hydroxide erosion assay: a revision of the alkali resistance test. AB - Burckhardt proposed the alkali resistance method as a means of assessing the integrity of the stratum corneum barrier in 1947. Researchers after Burckhardt largely found the test unreliable and nonreproducible; it therefore fell into disuse worldwide. We have upgraded the procedure by exposing the skin to 1.0 M sodium hydroxide under strictly specified conditions for successive 1-min periods until the emergence of the first erosions, revealed by staining with nitrazine yellow. Histology showed that the erosions were follicular and limited to the epidermis. The test was highly reproducible and repeatable. We demonstrated the usefulness of the test in the following ways: (1) the erosion time increased with aging, correlating with a thickened horny layer; (2) as few as five Scotch tape strippings greatly decreased the erosion time, although transepidermal water loss was only slightly increased; (3) slight damage to the horny layer by a 24-h exposure to 0.01% sodium lauryl sulfate sharply reduced the erosion time; (4) the erosion time decreased after daily open applications for 3 weeks of clobetasol propionate, corresponding to the thinned horny layer; (5) daily applications of petrolatum increased the erosion time. This new version of the alkali resistance test, renamed the sodium hydroxide erosion assay, promises to be a useful tool for studying the horny layer barrier in healthy and diseased skin. PMID- 9749994 TI - Lack of correlation of skin thickness with bone density in patients receiving chronic glucocorticoid. AB - The objective was to test the hypothesis that there is a correlation between thinning of the skin and bone in patients on chronic oral glucocorticoids (GCs). This was a one-time cross-sectional analysis performed in an academic referral center. The study group consisted of 14 patients on GCs for a variety of disorders, including dermatomyositis, pemphigus vulgaris, pyoderma gangrenosum, and urticarial vasculitis. Skin thickness was compared with that of 24 sex- and age-matched controls. The main outcome measures were the bone density of the lumbar spine (L2-L4) and the skin thickness. The skin thickness (mm, mean +/- SEM) in GC-treated (n = 7) vs unmedicated age-matched Caucasian women (n = 20) was 0.84 +/- 0.04 vs 1.02 +/- 0.04 (t = 3.07, P < 0.01) in the upper arm, 1.13 +/ 0.09 vs 1.49 +/- 0.05 (t = 3.65, P < 0.002) in the dorsal forearm, and 0.96 +/- 0.07 vs 1.17 +/- 0.02 (t = 2.92, P < 0.01) in the ventral forearm. L2-L4 bone densities averaged 106 +/- 2% in the GC-treated female patients relative to the age and sex-matched controls. There was no correlation between skin thickness and bone density. In GC-treated (n = 4) vs unmedicated Caucasian men matched for age (n = 4), skin thickness was 1.09 +/- 0.4 vs 1.33 +/- 0.05 (t = 3.51, P < 0.02) in the upper arm, but was not significantly different at the two forearm sites. No correlation between skin thickness and bone density was observed. The level of type I procollagen mRNA in skin from three GC-treated patients was 45% of the value in three age-matched controls. In conclusion, GCs cause statistically significant thinning of skin independently of the effects on bone. PMID- 9749995 TI - The reduced folate carrier (RFC-1) gene is expressed in the murine epidermis. PMID- 9749996 TI - The T-cell receptor Vbeta repertoire of nickel-specific T cells. PMID- 9749997 TI - Arsenic induces decreased expression of beta2-adrenergic receptors in cultured keratinocytes. PMID- 9749998 TI - Inhibition of 5-HT3 receptor cation channels by ifenprodil in excised patches of N1E-115 cells. AB - The patch-clamp technique was applied in out-side-out patches of N1E-115 mouse neuroblastoma cells to investigate the effects of ifenprodil [(+/-) erythreo ifenprodil tartratel, a drug with neuroprotective properties in cerebral ischemia, on the inward currents through 5-HT3 receptor channels. A high time resolution was achieved by using a rapid solution exchange system (exchange rate <1 ms). Ifenprodil inhibited the peak currents evoked by 30 microM 5-HT in a concentration-dependent but voltage-independent manner. The effect was most potent when ifenprodil was continuously applied to the patches 45 s before and during the 2-s administration of 5-HT (IC50=16 microM) and it was only slightly less potent when it was applied during the 45 s prior to 5-HT only (IC50=29 microM). When applied in this manner, ifenprodil also produced a concentration dependent increase of the onset time constant (tauON) of the 5-HT (30 microM) induced currents. When the drug was exclusively co-applied with 5-HT, ifenprodil was least potent in inhibiting the peak currents (IC50=98 microM), and it had no effect on the current onset kinetics. All protocols of ifenprodil application accelerated current inactivation as reflected by a decrease of the current inactivation time constant (tauOFF). All effects of ifenprodil were reversible after washout periods of 2-5 min. In conclusion, the potency of ifenprodil in inhibiting the inward current through 5-HT3 receptor channels is strongly dependent on the application protocol: presence of the drug before the agonist induced activation of the 5-HT3 receptor channels is necessary for a relatively potent inhibition of the 5-HT-induced peak current and is a prerequisite for the prolongation of tauON; in addition, a weak but fast inhibitory effect on the current amplitude and decay constant of the 5-HT-induced current was revealed by the experiments in which ifenprodil was exclusively present during exposure to 5 HT. Three alternatives compatible with the components of the ifenprodil effect have been discussed: (1) different effects of the two enantiomers, (2) action via two different mechanisms, and (3) operation via a single mechanism only. PMID- 9749999 TI - Inhibition of ATP-induced cAMP formation by 5'-p-fluorosulfonylbenzoyladenosine in NG108-15 cells. AB - ATP is known to increase intracellular cAMP levels in NG108-15 cells via a novel purinoceptor and this response is inhibited by the P1 purinoceptor antagonist methylxanthine. In the present study, we examined the effects of 5'-p fluorosulfonylbenzoyladenosine (FSBA), an affinity ligand for ATP-binding proteins, on cAMP formation mediated by activation of adenylate cyclase (AC) linked purinoceptors in NG108-15 cells. cAMP levels were determined by RIA using an anti-succinyl-cAMP antiserum. FSBA (100 microM) increased intracellular cAMP about 2.6-fold. However, FSBA-induced cAMP formation was abolished by pretreatment with adenosine deaminase, suggesting that adenosine, a breakdown product of FSBA, is involved in FSBA-induced cAMP formation. In contrast, pretreatment of cells with FSBA in the presence of adenosine deaminase inhibited cAMP formation induced by ATP and beta,gamma-methylene-ATP (beta,gamma-MeATP), without affecting the prostaglandin E1 (PGE1)-induced response. The inhibitory effect of FSBA on ATP-induced cAMP formation was concentration-dependent with a concentration required for half-maximal inhibition (IC50) of around 3 microM. The inhibitory effect of FSBA was not affected by pertussis toxin (PTX)-treatment. Pretreatment with FSBA (10 microM) depressed the maximal response to beta,gamma MeATP by 60%, but did not affect the response to 5'-N-ethylcarboxamidoadenosine. The inhibitory effect of FSBA (100 microM) increased time-dependently during pretreatment and partly resisted wash-out. The inhibition by FSBA was protected by simultaneous addition of beta,gamma-MeATP during the FSBA pretreatment, indicating that both FSBA and the ATP analogue interacted with the same receptor site. The pretreatment with FSBA did not affect the increase in [Ca2+]i induced by ATP, UTP or benzoylbenzoic ATP. These results suggest that FSBA inhibits cAMP accumulation induced in NG108-15 cells by ATP or related agonists by selective modification of an AC-linked purinoceptor. PMID- 9750000 TI - Analysis of a porcine EP3-receptor: cloning, expression and signal transduction. AB - A cDNA clone, encoding a complete porcine EP3 receptor, was isolated from a porcine heart cDNA library. The deduced amino acid sequence revealed a protein of 387 amino acid residues with an estimated molecular weight of 43 kD and strongest homology to the human EP3-II receptor (84% identity on protein level). Ligand binding studies with transfected COS-7 cells, expressing the porcine receptor, showed displacement of [3H]PGE1 with the EP3-specific agonist M&B 28.767, the EP1/EP3-agonist sulprostone but not with the EP2-specific agonist butaprost. Stimulation of transfected CHO cells with M&B 28.767 resulted in inhibition of forskolin-induced cAMP formation, suggesting coupling to an inhibitory G protein. Agonist-induced translocation of the transcription factor NFkappaB into the nucleus of transfected CHO cells was demonstrated by Western blot analysis, indicating that these EP3 receptors modulate NFkappaB-dependent cellular signal transduction. Analysis of the genomic organization identified the major transcription initiation site at about 160 bp upstream of the ATG start codon. The 800-bp 5' flanking region contains a variety of putative cis-acting regulatory elements, including binding sites for AP2, SP1 and MyoD (E-box). The present data will now allow further studies on EP3 receptor-mediated signal transduction and its regulation. PMID- 9750001 TI - Blockade of GABA(B) receptors facilitates muscarinic agonist-induced epileptiform activity in immature rat piriform cortex in vitro. AB - The effects of the selective GABA(B) receptor antagonist [3-[[(3,4 dichlorophenyl)methyl]aminolpropyl] (diethoxymethyl) phosphinic acid (CGP 52432) on muscarinic (mAChR) and metabotropic glutamate (mGluR) responsiveness were studied in slices of piriform cortex from both immature (P16-P22) and adult (> or =P40) rats, using a conventional intracellular recording technique. In both adult and immature slices, CGP 52432 (1 microM) had no effect on neuronal membrane properties, whereas it selectively abolished the late inhibitory postsynaptic potential (IPSP) evoked by local electrical stimulation of association fibre terminals. Age-related changes in mAChR (but not mGluR) responsiveness were also detected. In adult neurones, bath-application of the mAChR agonist oxotremorine-M (OXO-M; 10 microM), or the selective mGluR agonist 1S,3R-aminocyclopentane-1,3 dicarboxylic acid (1S,3R-ACPD; 10 microM) evoked similar membrane depolarization and inhibition of evoked excitatory postsynaptic potentials (EPSPs). However, while 1S,3R-ACPD and OXO-M produced indistinguishable slow excitatory effects in immature slices, during superfusion with OXO-M, neurones exhibited spontaneous paroxysmal depolarizing shifts (PDSs) that were suppressed in the presence of atropine (1 microM) or the selective GABA(B) receptor agonist beta parachlorophenyl-gamma-aminobutyric acid [(-)baclofen; 10 microM]. Also, application of OXO-M resulted in a pronounced prolongation (rather than a decrease) of electrically evoked postsynaptic potentials (PSPs) which now exhibited recurrent superimposed spike discharges. In adult slices, in the continuous presence of CGP 52432 (1 microM; 20 min pre-incubation), a subsequent exposure to 10 microM OXO-M or 1S,3R-ACPD failed to induce any spontaneous epileptiform activity, and evoked PSPs were consistently suppressed. In contrast, in immature slices, after incubation in CGP 52432 (1 microM; 20 min), a subsequent application of a low dose of OXO-M (2.5 microM), which was inactive per se, was able to produce spontaneous PDSs and a prolongation of evoked PSPs. We conclude that a reduction in GABA(B)-mediated synaptic inhibition in immature slices (in co-operation with other factors) may contribute to the facilitation of excitatory neurotransmission and therefore play a role in the generation of mAChR induced epileptiform activity. PMID- 9750002 TI - Potent, stereoselective, and brain region selective modulation of second messengers in the rat brain by (+)LY354740, a novel group II metabotropic glutamate receptor agonist. AB - LY354740 is a highly potent and selective agonist for recombinant Group II mGlu receptors (mGlu2 and mGlu3), which has anxiolytic and drug withdrawal alleviating properties when administered systemically in rats and mice. The modulation of second messengers by LY354740 in rat brain tissues was investigated to understand the cellular basis for the pharmacological and potential therapeutic actions of LY354740. LY354740 potently decreased forskolin-stimulated cAMP formation in slices of the adult rat hippocampus (EC50=22+/-3 nM) in a stereoselective manner. LY354740 (at 1 microM) greatly (>90%) suppressed forskolin-stimulated cAMP in the cerebral cortex, hippocampus, and striatum, while producing only partial suppression (about 50%) in midbrain regions and olfactory bulb, and no significant cAMP alterations in the cerebellum and brainstem regions. Inhibition of forskolin-stimulated cAMP formation was antagonized by (+)-alpha(-methyl-4 carboxyphenylglycine [(+)MCPG], a competitive mGlu receptor antagonist. LY354740 did not alter phosphoinositide hydrolysis in the rat hippocampus per se, but potentiated stimulation of phophoinositide hydrolysis by the Group I mGlu receptor selective agonist 3,5-dihydroxyphenylglycine (DHPG) or stimulation of cAMP formation by the adenosine receptor agonist 5'-N-ethylcarboxamideoadenosine (NECA). These data indicate that LY354740 is a highly potent, efficacious, and selective Group II mGlu receptor (mGlu 2/3) agonist in the rat brain. The potent, stereoselective, and brain region selective actions of LY354740 on mGlu receptor linked second messenger systems likely underlie the in vivo potency and stereoselectivity of this compound in animal models. PMID- 9750003 TI - Riluzole, a glutamate release inhibitor, and motor behavior. AB - Riluzole (2-amino-6-trigluoromethoxy benzothiazole) has neuroprotective, anticonvulsant, anxiolytic and anesthetic qualities. These effects are mediated by blockade of glutamate transmission, stabilizing of sodium channels and blockade of gamma-aminobutyric acid (GABA) reuptake. The action profile of riluzole is dominated by its effects on glutamate transmission which are predominately mediated by N-methyl-D-aspartate (NMDA) receptor-linked processes in vitro. In vivo studies show that blockade and stimulation of the different NMDA receptor complex binding sites or AMPA receptors modulate motor behavior in a characteristic manner. It was therefore interesting to examine if blockade of glutamatergic transmission by riluzole induced similar behavioral effects as direct NMDA/AMPA receptor antagonists and if these effects are mediated by a specific receptor. The effects of riluzole alone and in combination with several other neuroactive compounds on the central nervous system was assessed by behavioral paradigms to evaluate sniffing behavior, locomotion, ataxia and rigidity. Accompanying compounds included the NMDA receptor agonist NMDA, the partial glycine site agonist D-cycloserine (DCS), and the alpha-amino-3-hydroxy-5 phenyl-4-isoxazolepropionic acid (AMPA) receptor antagonist GYKI 52466 [1-(4 aminophenyl)-4-methyl-7,8-methylenedioxy-5H-2,3-benzo-diazepine HCl]. Riluzole influenced neither stereotyped sniffing behavior nor locomotion but impaired motor coordination and attenuated rigidity induced by blockade of dopamine D1 and D2 receptor antagonists when given alone. At higher doses spontaneous behavioral activity decreased and motor coordination was more impaired. Augmentation of the riluzole effects were observed when NMDA, but not GYKI 52466, was coadministered. The glycine site agonist DCS increased the anticataleptic properties of riluzole. The results indicate that when given alone, riluzole has a behavioral profile resembling that of competitive NMDA receptor antagonists. However, coadministration of riluzole with NMDA/AMPA receptor ligands suggests that this assumption is incorrect, and that riluzole affects glutamatergic transmission by a more indirect mechanism. Nevertheless, the profile of riluzole together with its pre- and postsynaptic blockade of glutamatergic transmission implies beneficial properties in diseases where an overactive glutamate system induces chronic neurotoxicity and/or acute behavioral effects. PMID- 9750004 TI - A novel Na+/Ca2+ channel blocker, NS-7, suppresses hypoxic injury in rat cerebrocortical slices. AB - The substance 4-(4-fluorophenyl)-2-methyl-6-(5-piperidinopentyloxy) pyrimidine hydrochloride (NS-7) has been developed recently as a cerebroprotective compound with Na+ and Ca2+ channel blocking action. In the present study, the effect of NS 7 in an in vitro model of hypoxic injury was examined and the possible involvement of Na+ and Ca2+ channels in the hypoxic injury subsequently determined. When slices of rat cerebral cortex were exposed to hypoxia/glucose deprivation followed by reoxygenation and restoration of the glucose supply, marked leakage of lactate dehydrogenase (LDH) occurred 3-6 h after reoxygenation. This hypoxia/reoxygenation-induced injury was blocked almost completely by the removal of extracellular Ca2+ or by chelating intracellular Ca2+ with 1,2-bis(o aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetra(acetoxymethyl)ester (BAPTA/AM). In addition, combined treatment with the N-type Ca2+ channel blocker omega-conotoxin GVIA and the P/Q-type Ca2+ channel blocker omega-agatoxin IVA significantly reduced LDH leakage, although neither of these Ca2+ channel blockers alone, nor nimodipine, an L-type Ca2+ channel blocker, was effective. On the other hand, several Na+ channel blockers, including tetrodotoxin, local anaesthetics and antiepileptics, significantly reduced the hypoxic injury. NS-7 (3-30 microM) concentration-dependently inhibited LDH leakage caused by hypoxia/reoxygenation, but had no influence on the reduction of tissue ATP content and energy charge during hypoxia and glucose deprivation. It is suggested that blockade of Na+ and Ca2+ channels is implicated in the cerebroprotective action of NS-7. PMID- 9750005 TI - Actions of alpha2 adrenoceptor ligands at alpha2A and 5-HT1A receptors: the antagonist, atipamezole, and the agonist, dexmedetomidine, are highly selective for alpha2A adrenoceptors. AB - This study examined the activity of chemically diverse alpha2 adrenoceptor ligands at recombinant human (h) and native rat (r) alpha2A adrenoceptors compared with 5-HT1A receptors. First, in competition binding experiments at h alpha2A and h5-HT1A receptors expressed in CHO cells, several compounds, including the antagonists 1-(2-pyrimidinyl)piperazine (1-PP), (+/-)-idazoxan, benalfocin (SKF 86466), yohimbine and RX 821,002, displayed preference for h alpha2A versus h5-HT1A receptors of only 1.4-, 3.6-, 4-, 10- and 11-fold, respectively (based on differences in pKi values). Clonidine, brimonidine (UK 14304), the benzopyrrolidine fluparoxan and the guanidines guanfacine and guanabenz exhibited intermediate selectivity (22- to 31-fold) for h alpha2A receptors. Only the antagonist atipamezole and the agonist dexmedetomidine (DMT) displayed high preference for alpha2 adrenoceptors (1290- and 91-fold, respectively). Second, the compounds were tested for their ability to induce h5 HT1A receptor-mediated G-protein activation, as indicated by the stimulation of [35S]GTPgammaS binding. All except atipamezole and RX 821,002 exhibited agonist activity, with potencies which correlated with their affinity for h5-HT1A receptors. Relative efficacies (Emax values) were 25-35% for guanabenz, guanfacine, WB 4101 and benalfocin, 50-65% for 1-PP, (+/-)-idazoxan and clonidine, and over 70% for fluparoxan, oxymetazoline and yohimbine (relative to 5-HT = 100%). Yohimbine-induced [35S]GTPgammaS binding was inhibited by the selective 5-HT1A receptor antagonist WAY 100,635. In contrast, RX 821,002 was the only ligand which exhibited antagonist activity at h5-HT1A receptors, inhibiting 5-HT-stimulated [35S]GTPgammaS binding. Atipamezole, which exhibited negligeable affinity for 5-HT1A receptors, was inactive. Third, the affinities for r alpha2A differed considerably from the affinities for h alpha2A receptors whereas the affinities for r5-HT1A differed much less from the affinities for h5-HT1A receptors. This affected markedly the affinity ratios of certain compounds. For example, (+/-)-idazoxan was only 3.6-fold selective for h alpha2A versus h5-HT1A but 51-fold selective for r alpha2A versus r5-HT1A receptors. Conversely, yohimbine was tenfold selective for h alpha2A versus h5-HT1A adrenoceptors but 4.2-fold selective for r alpha2A versus r5-HT1A receptors. Nevertheless, both atipamezole and DMT were highly selective for both rat and human alpha2A versus rat or human 5-HT1A receptors. In conclusion, these data indicate that: (1) the agonist DMT and the antagonist atipamezole are the ligands of choice to distinguish alpha2-mediated from 5-HT1A-mediated actions, whilst several of the other compounds show only low or modest selectivity for alpha2A over 5-HT1A receptors; (2) caution should be exercised in experimental and clinical interpretation of the actions of traditionally employed alpha2 ligands, such as clonidine, yohimbine and (+/-)-idazoxan, which exhibit marked agonist activity at 5-HT1A receptors. PMID- 9750006 TI - Prejunctional alpha2A-autoreceptors in the human gastric and ileocolic arteries. AB - This study was undertaken to determine the subtype of prejunctional alpha2 autoreceptors in human blood vessels. Segments of gastric and ileocolic arteries were incubated with [3H]noradrenaline and subsequently perifused with modified Krebs-Henseleit solution containing cocaine (12 microM). Five periods of electrical stimulation (S1-S5) were applied (1 Hz, 1 ms, 50 V for 1 min). Concentration-response curves for the facilitatory effect of eight alpha adrenoceptor antagonists [rauwolscine, 2-[2-(2-methoxy-1,4-benzodioxanyl)] imidazoline (RX821002), yohimbine, phentolamine, idazoxan, 2-(2',6' dimethoxyphenoxyethyl)-aminomethyl-1,4-benzodioxan (WB4101), spiroxatrine and prazosin] were determined. All antagonists enhanced the stimulation-evoked overflow of tritium, indicating the existence of alpha2-autoreceptors. The EC30% values of the antagonists (concentrations that increased the evoked overflow of tritium by 30%) were taken as a measure of affinity to the autoreceptors. Correlations between the pEC30% values obtained in the present study and the pKi values of the same antagonists at cloned human alpha2A-, alpha2B-, alpha2C adrenoceptors expressed in Chinese hamster lung cells and at alpha2D adrenoceptors in the rat submaxillary gland or the bovine pineal gland showed that the alpha2-autoreceptors in the human gastric and ileocolic arteries resemble most closely the alpha2A-subtype. PMID- 9750007 TI - Porcine carotid vascular effects of eletriptan (UK-116,044): a new 5-HT1B/1D receptor agonist with anti-migraine activity. AB - It has been suggested that opening of cephalic arteriovenous anastomoses may be involved in the headache phase of migraine. Indeed, a number of acutely acting anti-migraine drugs, including the ergot alkaloids and sumatriptan, constrict porcine carotid arteriovenous anastomoses. In this study, using pentobarbital anaesthetised pigs, we investigated the effects of eletriptan, a close structural analogue of sumatriptan, on the distribution of common carotid artery blood flow into arteriovenous anastomotic and nutrient (capillary) fractions. Eletriptan (10, 30, 100, 300 and 1000 microg kg(-1), i.v.) decreased the total carotid blood flow, exclusively by decreasing cephalic arteriovenous anastomotic blood flow; nutrient blood flow, particularly to the ear, skin and fat, was significantly increased. The doses of eletriptan needed to reduce arteriovenous anastomotic blood flow and conductance by 50% (ED50) were, respectively, 117+/-21 microg kg( 1) (251+/-45 nmol kg(-1)) and 184+/-42 microg kg(-1) (396+/-91 nmol kg(-1)); the highest dose caused reductions of 84+/-3% and 77+/-4%, respectively. The eletriptan-induced changes in carotid haemodynamics were clearly attenuated by pretreating the pigs with the selective 5-HT1B/1D receptor antagonist GR127935 (0.5 mg kg(-1)). On the basis of these results, we conclude that (1) the eletriptan-induced constriction of cephalic arteriovenous anastomoses as well as the arteriolar dilatation in head tissues is predominantly mediated by 5-HT1B/1D receptors, and (2) eletriptan should be effective in aborting migraine headache. Clinical studies have already demonstrated its therapeutic action in migraine patients. PMID- 9750008 TI - A study on possible modulating and direct effects of gamma2-MSH and ACTH-(1-24) on the cardiovascular system of the rat. AB - In conscious rats, gamma2-melanocyte-stimulating hormone (gamma2-MSH) dose dependently increases blood pressure and heart rate, whereas adrenocorticotropin (1-24) [ACTH-(1-24)] dose-dependently decreases blood pressure, an effect which was accompanied by a reflectory tachycardia. As the exact mechanism involved in these cardiovascular effects of the two melanocortins is as yet not known, we undertook a series of experiments to investigate the possibility that these peptides have modulating or direct effect on the cardiovascular system of the rat. In pithed rats gamma2-MSH, administered intravenously (i.v.) in doses of 5 200 nmol/kg, had no significant effect on systolic and diastolic blood pressure and on heart rate, whereas ACTH-(1-24), 5-500 nmol/kg, i.v., dose-dependently decreased blood pressure and increased heart rate. Infusion of gamma2-MSH, 10(-8) M, or ACTH-(1-24), 10(-6) M, in the isolated perfused rat heart did not significantly affect left ventricular pressure or coronary flow. Pretreatment with either gamma2-MSH or ACTH-(1-24) did not modify the responsiveness of the myocardium and coronary vasculature to salbutamol and phenylephrine. Neither gamma2-MSH nor ACTH-(1-24) did affect the vascular contractile machinery of skinned vascular smooth muscles of the rabbit with respect to Ca2+ handling in the cell, as measured by its sensitivity to exogenously applied Ca2+. Gamma2-MSH had no effect on blood pressure and heart rate in pithed rats in which postganglionic sympathetic outflow was stimulated by 1,1-dimethyl-4 phenylpiperazinium (DMPP), nor in pithed rats in which preganglionic sympathetic outflow was stimulated electrically. A dose of 15 nmol/kg ACTH-(1-24) had no significant influence on preganglionic outflow to the cardiac and vascular structures in pithed rats. These data show that gamma2-MSH does not exert its cardiovascular effects via a peripheral site of action at the level of the vascular system and the heart, nor directly on pre- or postganglionic sympathetic outflow. These results are in support for the notion that the peptide acts via a brain region localised outside the blood-brain barrier. The acute depressor effect of ACTH-(1-24), however, seems to be due to a direct effect on the vasculature in the periphery. PMID- 9750009 TI - Interaction of the diuretics torasemide and U-37883A with the K(ATP) channel in rat isolated aorta. AB - In this study we have investigated the interaction of the loop diuretics torasemide and furosemide and of the eukalemic diuretic U-37883A (4 morpholinocarboximidine-N-1-adamantyl-N'-cyclohexylhyd rochloride) with the ATP sensitive K+ channel (K(ATP) channel) in rat aortic rings. Torasemide contains a sulphonylurea group which might enable the compound to interfere with K(ATP) channels; this group is lacking in furosemide. U-37883A blocks several types of K(ATP) channels. The interaction with the vascular K(ATP) channel was probed in binding studies, 86Rb+ efflux experiments and vasorelaxation assays. Torasemide inhibited the binding of the K(ATP) channel inhibitor [3H]glibenclamide and of the opener [3H]P1075 with IC50 values of 19 and 45 microM, respectively; furosemide and U-37883A were inactive or interfered with binding in a nonspecific way. In 86Rb+ efflux experiments, the loop diuretics, at microM concentrations, inhibited basal tracer efflux to 50% whereas U-37883A had no effect. P1075 stimulated 86Rb+ efflux, a qualitative measure of K(ATP) channel opening, was inhibited by U-37883A and torasemide with IC50 values of 0.06 and 130 microM, respectively; furosemide induced only a small (23%) inhibition. In experiments measuring isometric force, torasemide and furosemide partially relaxed endothelium-denuded aortic rings precontracted with noradrenaline or KCl with EC50 values between 6 and 10 microM. The vasorelaxant effect of P1075 was inhibited in a noncompetitive manner by torasemide (300 microM) but unaffected by furosemide. U-37883A increased noradrenaline-induced force and inhibited the vasorelaxant effect of P1075 in an apparently competitive manner with an inhibition constant of 0.4 microM. The data show that torasemide interferes specifically with the binding of the K(ATP) channel modulators [3H]glibenclamide and [3H]P1075 and with the K(ATP) channel opening and vasorelaxant effects of P1075 whereas furosemide is inactive. This suggests that the interaction of torasemide with the vascular K(ATP) channel is due to the sulphonylurea group present in torasemide. U-37883A, which does not inhibit P1075 binding, is one of the most potent blockers of P1075-induced 86Rb+ efflux yet described but is relatively weak as an inhibitor of P1075-mediated vasorelaxation. The opposite vascular actions of torasemide and U-37883A are expected to contribute to the renal effects of these drugs. PMID- 9750010 TI - Eukaliuric natriuresis and diuresis in response to disprocynium24: studies on the tubular site of action. AB - We previously described that 1,1'-diisopropyl-2,4'-cyanine (disprocynium24, DP24) exerts an eukaliuric diuresis and natriuresis in the anesthetized rat. The purpose of the present study was to localize the tubular site of action of DP24. Employing micropuncture experiments in anesthetized rats, we first tested the effect of systemic application of DP24 (300 microg/kg + 300 microg/kg h, i.v.) on whole kidney excretion rates as well as on fluid, sodium and potassium ion delivery to the early distal tubule (V(ED), Na+(ED), K+(ED)). It was found that the eukaliuric diuresis and natriuresis in response to DP24 was accompanied by a substantial increase in V(ED) and Na+(ED), suggesting a predominant tubular site of action upstream to the early distal tubule, most likely in the proximal tubule. DP24 caused a comparable fractional, although minor absolute increase in K+(ED) as compared to Na+(ED). Second, application of DP24 into the first surface loop of the proximal tubule significantly increased V(ED) and Na+(ED) at a concentration of about 10(-7) M, indicating that DP24 may act from the intratubular site. Third, microperfusion of tubular segments revealed that effects of DP24 on the proximal convoluted tubule and the loop of Henle accounted for about 70 and 30%, respectively, of its diuretic and natriuretic action upstream to the early distal tubule. With regard to the loop of Henle, the quantitative effect of DP24 on fluid and Na+ reabsorption proposed a predominant effect on the straight part of the proximal tubule rather than the thick ascending limb. Intratubular DP24 did not affect reabsorption in the distal tubule. In summary, the present findings indicate that: (1) the diuretic and natriuretic effect of DP24 resides predominantly in the proximal tubule, and (2) DP24 may act from the intratubular site. Since DP24 increased V(ED) and Na+(ED) without apparently affecting sodium or potassium ion transport in the distal tubule, the mechanism of the eukaliuric response remains unclear. PMID- 9750011 TI - Potassium diet as a determinant for the renal response to systemic potassium channel modulation in anesthetized rats. AB - The role of potassium intake in the response of kidney function and plasma renin activity (PRA) to systemic application of U37883A (4-morpholinecarboximidine-N-1 adamantyl-N'-cyclohexyl-hydro chloride), a putative blocker of ATP-sensitive potassium channels (K(ATP)), and P1075 (N-cyano-N'-(1,1-dimethylpropyl)-N" pyridylguanidine), an opener of K(ATP) channels, was studied in the anesthetized rat. It was found that under normal potassium diet (0.7% K), U37883A (15 mg/kg, i.v.) increased urinary flow rate (UV) and sodium excretion (UNaV), decreased urinary potassium excretion (UKV), and significantly diminished heart rate (HR) without affecting mean arterial blood pressure (MAP) or glomerular filtration rate (GFR). P1075 (10 microg/kg, i.v.) lowered UV, UNaV and UKV, at least in part due to the fall in MAP and GFR. PRA was diminished by U37883A and increased by P1075. Variation in potassium diet (0.04 or 2% K) left the response in MAP, HR or GFR to both potassium channel modulators essentially unchanged. The reduction in renal excretion rates to P1075 also appeared unaffected, further supporting a predominant role of the change in MAP and GFR in this response. Variation in potassium diet, however, elicited the following alterations: (1) under both low and high potassium diet U37883A did no longer cause a significant natriuresis; (2) U37883A elicited a significant kaliuresis under high potassium diet, whereas potassium excretion remained essentially unchanged on very low levels under low potassium diet; (3) the increase in PRA to P1075 was blunted under low potassium diet. Additional experiments provided evidence that P1075 releases renin from freshly isolated juxtaglomerular cells of rats on normal but not on low potassium diet. In summary, systemic potassium channel modulation employing U37883A or P1075, respectively, exerts distinct effects on blood pressure and heart rate independent of potassium diet. In contrast, potassium diet appears to be a determinant for the concomitant responses in plasma renin activity and renal sodium and potassium excretion. PMID- 9750012 TI - Protective effects of a PAF receptor antagonist and a neutrophil elastase inhibitor on multiple organ failure induced by cerulein plus lipopolysaccharide in rats. AB - The inhibitory effects of YM264, a selective platelet activating factor (PAF) receptor antagonist, and 2-(3-methylsulfonylamino-2-oxo-6-phenyl-1,2-dihydro-1 pyridyl)-N-( 3,3,3-trifluoro-1-isopropyl-2-oxopropyl)acetamide (compound 1), a neutrophil elastase inhibitor, on mortality, and pancreatic, hepatic, renal and pulmonary dysfunction were evaluated in a rat model of multiple organ failure (MOF) accompanying acute pancreatitis. MOF was produced by intraperitoneal injection of lipopolysaccharide (LPS, 30 mg/kg) in rats with cerulein-induced pancreatitis. LPS dose-dependently increased the mortality in rats with or without pancreatitis. The threshold dose which produced death in rats without pancreatitis was 30 mg/kg. This same dose evoked death in more than 40% of rats with pancreatitis. Time-course changes in serum enzyme and organ myeloperoxidase (MPO) levels were first examined in rats with induced MOF, and the results were compared with those in rats treated with only LPS or cerulein. Pancreatic weight, and serum amylase and lipase levels significantly increased in rats with cerulein induced pancreatitis despite the presence or absence of LPS, but recovery of these pancreatic dysfunctions was slower in the group given LPS. However, serum GOT, GPT, BUN and creatinine levels were significantly elevated only in MOF rats. In the MOF rats, the MPO level in the lung was significantly elevated and arterial oxygen pressure was decreased, indicating that infiltration of neutrophils into the lung might be involved in pulmonary dysfunction. However, the MPO levels in the pancreas and kidney in the MOF rats were not remarkably different from those in normal rats. The inhibitory effects of YM264 and compound 1 on mortality and organ dysfunction were examined in this MOF model. The 24-h survival rate for rats prophylactically and therapeutically treated with an intravenous infusion of YM264 at 0.1 mg/kg h was significantly higher than that of controls. The 24-h survival rate for rats treated prophylactically by intravenous infusion of 2 mg/kg h of compound 1 was significantly higher than that of control, whereas a beneficial dose of compound 1 was 5 mg/kg h in therapeutically treated rats. Prophylactic treatment with YM264 (0.1 mg/kg h) and compound 1 (2 mg/kg h) ameliorated organ dysfunction in rats with MOF. In conclusion, pancreatic, hepatic, renal and pulmonary dysfunctions are observed in this rat MOF model. The PAF receptor antagonist and neutrophil elastase inhibitor reduce the mortality rate in rats with MOF due to their inhibitory effects on organ dysfunction, indicating that PAF and neutrophil elastase may play important roles in the development of MOF. These results in the present model are largely consistent with those in patients with MOF, indicating that this model is suited for MOF in humans and may be used as a model to test new therapeutic approaches. PMID- 9750013 TI - Effect of colchicine on nerve growth factor-induced leukocyte accumulation and thermal hyperalgesia in the rat. AB - In addition to its neuronal effects, nerve growth factor (NGF) is known to act on inflammatory and immune cells. The aim of the present study was to investigate the effect of colchicine on NGF-induced leukocyte accumulation and thermal hyperalgesia. Initial experiments showed that intradermal injection of recombinant human (rh) NGF (0.8 and 4 microg) caused a longlasting increase in tissue myeloperoxidase (MPO) indicating leukotactic activity of NGF. Colchicine (0.3 and 1 mg/kg) attenuated the NGF (0.8 and 4 microg)-induced increase in tissue myeloperoxidase (MPO) as determined 6 h after NGF application. Intraplantar injection of NGF into the rat hindpaw caused a decrease in thermal nociceptive threshold, which, at 4 microg NGF, was accompanied by moderate (about 35% increase in paw volume) edema. The thermal hyperalgesia was evident 20 min after injection and lasted less than 4 h. Colchicine (0.3 and 1 mg/kg) had no significant effect on NGF-induced edema, but reduced NGF-induced thermal hyperalgesia. Colchicine (1 mg/kg) did not significantly reduce thermal hyperalgesia produced by intraplantar bradykinin, prostaglandin E1, or 5 hydroxytryptamine. Treatment of rats with a dose of indometacin (2 mg/kg) that was sufficient to block cyclooxygenase had no significant effect on NGF-induced thermal hyperalgesia or edema. In vitro, colchicine (0.4-12 microg/ml) did not significantly influence NGF (10 ng/ml)-induced histamine release from rat peritoneal cells, suggesting that a mast cell stabilizing effect of colchicine did not contribute to inhibition of NGF-induced thermal hyperalgesia. The results show that NGF causes localized indometacin-resistant thermal hyperalgesia that can be blocked by the microtubule disrupting agent colchicine. These results raise the possibility that a mechanism by which NGF produces peripheral sensitization is related to its leukotactic effect. PMID- 9750014 TI - Effects of NMDA receptor channel blockers, dizocilpine and memantine, on the development of opiate analgesic tolerance induced by repeated morphine exposures or social defeats in mice. AB - Development of tolerance to opiates involves various neurochemically and pharmacologically distinct processes. For instance, the diversity of opiate tolerance has been suggested by experiments comparing the establishment of diminished response to different effects of opiate agonists. Antagonists acting at N-methyl-D-aspartate (NMDA) receptors has become a very useful tool for studying opiate tolerance mechanisms since these drugs have been shown to retard the development of tolerance to analgesic properties of opiates. The present study compared the ability of two NMDA receptor channel blockers, dizocilpine and memantine, to affect the development of tolerance to morphine analgesia induced by repeated social defeat or by repeated morphine administrations. Male BALB/c mice were assessed for the tail-flick response before and after the defeat in five social confrontations, or before and after repeated morphine injections (20 mg/kg, s.c., once daily for 8 days). Repeated morphine injections were explicitly paired with environmental cues. Socially-defeated as well as morphine-treated mice developed significant tolerance to morphine analgesia. Separate groups of mice were exposed to repeated social confrontations or injections of morphine with each defeat or morphine injection followed by administration of either dizocilpine (0.03-0.3 mg/kg, i.p.) or low-affinity channel blocker memantine (3 30 mg/kg, i.p.). Both dizocilpine and memantine were effective in preventing the development of repeated morphine-induced tolerance to acute morphine analgesia. Treatments with NMDA receptor antagonists that retarded development of non associative tolerance also suppressed the establishment of associative tolerance significantly. Social defeat-induced tolerance was prevented by dizocilpine but not by memantine. Our results suggest some degree of similarity in the mechanisms of morphine analgesic tolerance induced by pharmacological, contextual and social stimuli. PMID- 9750015 TI - The inhibitory effect of vitamin E on 4-(methylnitrosamino)-1-(3-pyridyl)-1 butanone-induced DNA injury and the fixation of the DNA injury in mouse lungs. AB - In order to estimate the effect of vitamin E on DNA injury and K-ras point mutation at an early stage of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone(NNK) induced lung tumorigenesis in mice, the present study was carried out. Presupplement with vitamin E about 15 times more than control for a week significantly inhibited NNK-induced O6-methylguanine formation in the lungs of mice at 4 and 168 h after the injection. At 30 days after the NNK injection. the activation of K-ras oncogene with a 12th codon GC-->AT transition was detected in 56% of lung samples tested by mutant-allele-specific amplification. Vitamin E supplement reduced the frequency of the mutation to 30%. These results suggest that vitamin E suppresses NNK-induced DNA injury and subsequent fixation of the injury during the initiation and post-initiation phases of the lung tumorigenesis in mice. PMID- 9750016 TI - Long recovery times improve the detection of cellular resistance in vitro. AB - In vitro cytotoxicity testing is used increasingly during the development of clinical treatment protocols. These tests are influenced by many variables, not all of which have been assessed systematically yet. We analyzed the influence of the recovery time between the end of treatments and measurements on the detection of cellular resistance. The development of resistance to cisplatin and radiation was chosen as a model since the schedule of these treatments is the objective of several ongoing clinical trials. C6 rat glioma, T98G, 86HG-39, A172 human glioma and TE671 human rhabdomyo-sarcoma cells were pretreated with radiation or cisplatin. The cellular resistance was then compared in pretreated and wild-type cells using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test. In all cell lines, apparent drug concentrations killing 50% of the cells were dependent on the recovery time. In A172 cells this concentration was 10.3+/-02.1 microM after 48 h but decreased to 3.56+/-0.44 microM after 120 h recovery time (P < 0.001). After recovery times of more than 168 h, 53% of all pretreated cell lines were resistant to cisplatin or radiation, 25% were unchanged and 22% were more sensitive. However, only half the resistant cells could be identified when the MTT test was done with only 48 h recovery time. The sensitivity of detection increased from 0.46 to 0.83 when the recovery time of the test system was extended from 48 h to 168 h. The specificity was not dependent on the recovery time. Experiments showing resistance after short recovery times are reliable, but lack of resistance can only be shown in experiments with long recovery times. Cisplatin treatment can result in resistance to radiation in glioma cells. PMID- 9750017 TI - Radiosensitivity of dermal and tumor fibroblasts derived from the same patient. AB - The in vitro radiosensitivity of dermal fibroblasts has been found to vary between individuals, and a number of studies have also shown that this parameter correlates with radiation-induced late injuries in clinical radiotherapy. In addition, certain genetic disorders are known to effect radiosensitivity, e.g. normal tissues of patients homozygous or heterozygous for the ataxia teleangiectasia gene show unusual sensitivity to radiation both in vivo and in vitro. Thus, it has been assumed that there is a genetically determined component resulting in a certain intrinsic cellular radiation response in an individual. To study this possible relationship between different cells of a specific patient, we established eight pairs of dermal and tumor fibroblast cultures. The donor patients had either adenocarcinoma of the uterus or squamous cell carcinoma (SCC) of the head and neck. The radiosensitivity of these strains was determined by a 96-well plate clonogenic assay, previously used by us for radiosensitivity testing of cancer cells. From a paired comparison, the values for the cell fraction surviving 2.0 Gy (SF2), of both fibroblast strains, were found to be on the same level in five out of eight cases. In patient 6, the SF2 of tumor fibroblasts was significantly higher than that of dermal fibroblasts (P=0.0014). In two additional cases the tendency was the same, but not statistically significant. As groups, the two types of fibroblasts did not differ from each other, mean SF2 values of 0.24+/-0.07 and 0.21+/-0.05, respectively. The SF2 of tumor fibroblasts from SCC patients proved to be significantly higher than that of the adenocarcinoma patients (P=0.030). These preliminary results indicate that the in vitro radiosensitivity of tumor fibroblasts correlates with normal cell sensitivity in many cases, but not in all. The radiosensitivity of tumor fibroblasts also seems to follow the level of in vitro radiosensitivity determined for the corresponding histological type of tumor cells. Further studies are needed to determine more closely the relationship between the radiosensitivities of tumor cells and tumor fibroblasts, thus evaluating the possibility of testing radiosensitivity from tumor fibroblasts in order to estimate tumor response. PMID- 9750019 TI - Determination of growth fraction and cell density to evaluate the potential growth of human oligodendroglial and astrocytic tumours. AB - The object of this work was PURPOSE: to develop a methodology that enables net tumour growth, a balance between actual tumour growth and tumour cell loss, to be approximately evaluated. METHODS: The methodology proposed relies on detecting the growth fraction immunohistochemically by means of MIB-1 antibody labelling combined with cell density determination, carried out on 5-microm-thick Feulgen stained histological sections with computer-assisted microscopy. The series investigated included 25 oligodendrogliomas (OLG-II), 9 anaplastic oligodendrogliomas (OLG-III). 13 astrocytomas (AST), 14 anaplastic astrocytomas (ANA) and 8 mixed oligoastrocytomas (OLG-AST). RESULTS: The results show that the biological characteristics of some cases were in total accordance with their histopathological diagnoses. This was the case for the "weakly proliferating weakly dense" OLG-II and AST-II tumours, and for the "highly proliferating highly dense" OLG-III and AST-III ones. In contrast, the biological characteristics of some cases seemed to contradict the histopathological case labels. This was the case for the "highly proliferating highly dense" OLG-II and AST-II tumours, the biological aggressiveness of which would be undervalued on the basis of the morphology-based grading system alone, and also for the "weakly proliferating weakly dense" OLG-III and AST-III tumours, the aggressiveness of which would be overvalued. CONCLUSIONS: Combining the determinations of the MIB-1 and the cell density variables appears to be satisfactory in terms of the cell kinetic characterization of glial tumours as a complement to the prognostic information given by a morphology-based grading system alone. PMID- 9750018 TI - Absence or decreased levels of the hMLH1 protein in human gastric carcinoma cell lines: implication of hMLH1 in alkylation tolerance. AB - Defective hMLH1 function has been increasingly associated with acquired cellular resistance to DNA alkylation damage in human colorectal and endometrial cancer cells. To investigate the relationship between the DNA alkylation tolerance and the hMLH1 status in human gastric carcinoma cells, we determined the cellular response to N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), the mutational changes, and the expression of hMLH1 in 11 human gastric carcinoma cell lines. Of 11 cell lines, 4 (SNU-5, -16, -620, and -719) were sensitive, whereas 7 (SNU-1, -216, 484, -520, -601, -638, and -668) were resistant to the cytotoxic effect of MNNG. As determined by Western analysis, it was evident that all the MNNG-resistant cell lines except one (SNU-601) produced very low or undetectable levels of hMLH1 protein compared to the MNNG-sensitive cell lines. A homozygous non-sense mutation that resulted in truncated protein was found in one MNNG-resistant cell line (SNU-1). Therefore, to determine whether the sensitivity of cells to MNNG can be restored by exogenous expression of hMLH1 protein, wild-type hMLH1 cDNA was introduced into the MNNG-resistant cells (SNU-1). The cytotoxicity test showed that expression of exogenous wild-type hMLH1 protein caused an increase in sensitivity to the cytotoxic effect of MNNG. This restoration was confirmed by an increase in the cell population containing less than the G1 amount of DNA (cell death) in the wild-type hMLH1-transfected cells, as determined by flow cytometry analysis. Together our results suggest that (1) the absence or decreased level of wild-type hMLH1 protein may be a frequent event in the human gastric carcinoma cell lines, (2) such alterations in the hMLH1 protein are closely associated with the MNNG tolerance in the human gastric carcinoma cell lines, and (3) the hMLH1 protein participates not only in the repair of DNA mismatches but also in the mechanism of escape from the cytotoxic effects of DNA alkylation damage. PMID- 9750020 TI - Production and pre-clinical significance of haematopoietic peptide growth factors (HPGF) in human non-seminomatous germ cell tumour cell lines. AB - Peptide growth factors involved in the regulation of haematopoiesis (HPGF), for example granulocyte-colony-stimulating factor (G-CSF) and granulocyte/macrophage colony-stimulating factor (GM-CSF), are of clinical importance in the treatment of testicular germ cell tumour (GCT) patients with modern chemotherapy regimens since they ameliorate chemotherapy-induced neutropenia. Aberrant expression of and/or response to HPGF has been reported in several solid tumour types although no data are available on GCT with the exception of those on stem cell factor (SCF). The aims of this pre-clinical study were twofold: (1) to screen a panel of human non-seminomatous (NS)GCT for the production of HPGF and (2) to test the effects of G-CSF or SCF on the growth of NSGCT cell lines in vitro, and on the growth kinetics of two human NSGCT xenograft models. HPGF concentrations in cell culture supernatant from 11 NSGCT cell lines growing under routine culture conditions were measured by enzyme-linked immunosorbent assay. The growth kinetics of cell lines was quantified in vitro using the sulphorhodamine B assay. The growth kinetics of nude mouse NSGCT xenografts was followed by measuring tumour volumes every 2-3 days over days 1-30, following daily subcutaneous injection of nude mice (days 1-14). The cell lines produced G-CSF (1/11 cell lines), GM-CSF (2/11), SCF (2/11), M-CSF (6/11). and interleukin-6 (9/11). Growth stimulation of cell line H12.1 by SCF was observed in vitro, but no statistically significant differences in NSGCT xenograft tumour volume (VT) or relative VT (r VT) in treated groups were observed on days 14 or 29 compared to the control. The change in rVT of H12.1 xenografts treated with G-CSF alone compared to control (rVT/rVT,c) was 0.96 on day 29. The values for rVT/rVT,c for H12.1 xenografts treated with G-CSF in combination with low- or high-dose SCF were, respectively, 1.67 or 1.7 compared to 1.19 for SCF-treated mice. The results are in agreement with clinical data to date where no observations have been reported of stimulation or inhibition of tumours in patients receiving treatment with G-CSF. Before any clinical trials are initiated in GCT patients treated with G-CSF in combination with SCF, further pre-clinical experiments with this tumour type are recommended to investigate this phenomenon further in a greater number of NSGCT cell lines in vitro and in vivo and with a wider range of SCF/G-CSF schedules. The potential relevance of secretion of HPGF in NSGCT cell lines in vitro to the pathobiology of GCT in patients is also a subject of interest for future research. PMID- 9750021 TI - Telomerase activity significantly correlates with cell differentiation, proliferation and lymph node metastasis in colorectal carcinomas. AB - Telomerase activity was examined by the telomeric repeat amplification protocol assay, in a total of 37 colorectal adenocarcinomas, including stages A, B and C according to the Astler and Collier classification, and correlated with clinicopathological features. Of 17 stage C lesions, 13 were positive (76.5%; P<0.01), demonstrating a significant correlation with lymph node metastasis. In contrast, only 6 of 20 stage A and B carcinomas were positive (30.0%), this being significantly lower (P < 0.05). Moderately or poorly differentiated subtypes were more predominant in the telomerase-positive than in the telomerase-negative groups (P< 0.05) with greater elevation of mitotic and Ki-67 labeling indices (P < 0.0001). No significant relation was found between telomerase activity and p53 protein accumulation or Bcl-2 protein expression. The good correlation with tumor staging, lymph node metastasis, differentiation, and mitotic and Ki-67 labeling indices suggests that this parameter might have potential application in estimation of prognosis. PMID- 9750022 TI - The role of serum and gastric juice levels of carcinoembryonic antigen, CA19.9 and CA72.4 in patients with gastric cancer. AB - PURPOSE: The aim of the present study was to investigate carcinoembryonic antigen (CEA), CA19.9, and CA72.4 in the serum and gastric juice of patients with gastric cancer. METHODS: Serum and gastric juice tumor markers CEA, CA19.9, and CA72.4 were measured in 59 patients who had gastric adenocarcinomas and were undergoing curative gastrectomy. The same markers were measured in 47 patients with benign gastric disorders and in 40 healthy subjects. The correlation between the serum and gastric juice levels of tumor markers and several clinicopathological factors were evaluated by univariate analysis. The significance of the tumor markers as prognostic factors was assessed both by univariate and multivariate analysis. RESULTS: The positivity rates of serum CEA, CA19.9, and CA72.4 were 57.6%, 38.9%, and 18.6% respectively. The positivity rates of gastric juice CEA, CA19.9, and CA72.4 were 62.7%, 30.5%, and 23.7% respectively. The combination of serum and gastric juice markers gave a positivity of 81.3%. There was no correlation between serum and gastric juice level of each tumor marker. Positivity of gastric juice markers did not correlate with prognosis. A significant difference in prognosis was observed between patients positive and negative for serum CEA and CA19.9. Multivariate analysis also revealed that serum CEA and CA19.9 levels were independent prognostic factors. CONCLUSIONS: Levels of both serum and gastric juice tumor markers continue to have only limited diagnostic usefulness in gastric cancer patients. CEA and CA19.9 in the preoperative sera are good prognostic factors, whereas the presence of tumor markers in the gastric juice does not play any prognostic role. PMID- 9750024 TI - Cell signalling and cancer treatment. AACR special conference in cancer research in collaboration with the British Association for Cancer Research, the German Cancer Society (Section for Experimental Cancer Research), the Austrian Biochemical Society, and the Austrian Cancer Society. 23-28 February 1997, Telfs Buchen, Austria. PMID- 9750023 TI - Tumour-proliferative fraction and growth factor expression as markers of tumour response to radiotherapy in cancer of the uterine cervix. AB - This study seeks to define the role of pretreatment of evaluation of tumour growth fraction in cervical cancer and its relationship to the clinical course of the disease. In addition, it also seeks to explain whether cell kinetics and growth factor expression have an association with tumour response to radiotherapy and hence could be of value in the management of patients. All pre-treatment biopsies were analysed for the tumour-proliferative compartment by evaluation of Ki67 antigen expression and argyrophilic nucleolar organiser region (AgNOR) counts. Growth factor analysis was done by analysing for expression of epidermal growth factor (EGF), epidermal growth factor receptor (EGF-R) and transforming growth factors alpha and beta (TGFalpha, TGFbeta). A total of 152 patients were evaluated and a correlation obtained between pre-treatment status of the tumour growth-fraction-associated markers and clinical outcome following radiotherapy. Such patients were either disease-free (group 1, n=106) or with residual/recurrent disease (group 2, n=46) at a 16-month follow-up. Pre-treatment analysis of AgNOR significantly correlated to disease status after treatment (r= 0.517, P=0.0000). This may be due to an effect of cell proliferation. Lower AgNOR counts were significantly associated with recurrent/residual tumours, suggesting that increased proliferative activity may be a positive prognostic indicator. Similar results were also obtained for the other proliferation-associated marker Ki67 (r=-0.443, P=0.0000). Expression of EGF and EGF-R also showed significant pre-treatment correlations with the final disease outcome (r=0.248, P=0.031 and r=0.503, P=0.0000 respectively). Both these markers were expressed more by patients belonging to group 2. The opposite was the case for TGFalpha, where patients belonging to group 1 showed higher values (r=0.417, P=0.0001). The other growth factor investigated, TGFbeta, also showed a conspicuous differential expression in the two groups of patients (r=-0.604, P=0.0000). Group 1 patients showed mostly mild to moderate expression while most group 2 patients were negative for the growth factor. It therefore appears that tumours with high AgNOR counts and Ki67 index, along with expression of the two types of transforming growth factor (alpha and beta), responded better to radiotherapy. PMID- 9750025 TI - Clinical evaluation of biologically targeted drugs: obstacles and opportunities. AB - Recent insights into the molecular mechanisms of cancer have indicated that a variety of fundamental cellular processes are dysregulated in malignant cells. These processes include cell cycle control, signal transduction pathways, apoptosis, telomere stability, angiogenesis, and interactions with the extracellular matrix. Remarkable advances in molecular genetics, enzymology, and medicinal chemistry have permitted the design of compounds that modulate some of these processes with specificity that was unimaginable a decade ago. As these novel, biologically targeted compounds enter the clinic, they will require a strategy for clinical evaluation and development different from that used commonly for cytotoxic antineoplastic agents. This review examines the development of cancer drugs directed against angiogenesis, metastasis, signal transduction, telomerase, and molecular message (antisense), outlines strategies for the clinical testing of agents directed at these processes, and contrasts these efforts with traditional approaches to cancer drug testing. PMID- 9750026 TI - Overview of recent topics in clinical pharmacology of anticancer agents. AB - The rationale for studying the clinical pharmacology of antineoplastic agents is that the information obtained will result in enhanced drug development and enhanced or improved clinical use. A great deal of effort has been expended in studying the pharmacokinetics and pharmacodynamics of investigational and noninvestigational antineoplastic agents. More recently, a deeper appreciation has developed regarding the importance of the metabolism of antineoplastic agents and the potential role of metabolites in their activity or toxicity, as well as the potential for drug-drug interactions. Investigators studying the clinical pharmacology of antineoplastic agents face an increasingly challenging task as new agents continue to be developed. Some of these challenges arise from the enhanced potency of new agents, resulting in increased difficulty in measuring such agents in biological matrices. Furthermore, as agents have been developed to affect specific biological targets, the necessity of assessing pharmacodynamics at the biochemical or molecular level has become increasingly important. In addition, development of agents with cytostatic, as opposed to cytotoxic, properties poses a further challenge to assessment of pharmacologic effect. In addressing these challenges, a great deal of effort has been expended to develop increasingly sensitive analytical chemical techniques, in evaluating alternative biological matrices, such as saliva, in which to monitor drug concentrations in a less invasive fashion, and in developing limited sampling strategies to assess both the pharmacokinetics and pharmacodynamics of antineoplastic agents. Similarly, a great deal of effort has been expended in providing suitable means for assessing the numerous novel targets for which antineoplastic agents are being developed. These include the assessment of cell cycle kinetics and specific oncoproteins, definition of cell damage such as cleavable complexes, and formation of drug-macromolecular adducts in suitable target cells. Additional effort is being expended to explore nontraditional means of drug delivery. In this regard, the increasing importance of orally administered agents reflects a fundamental change in the approach to antineoplastic drug delivery. Finally, the increased computational power made available by faster personal computers has facilitated a number of innovative modeling techniques involving population modeling, modeling of combination chemotherapy, and assessment of drug-drug interactions. PMID- 9750027 TI - Clinical pharmacology of camptothecins. AB - Camptothecins (CPTs) are a unique class of chemotherapeutic agent which inhibit DNA synthesis by inhibiting topoisomerase I activity. Structure-activity studies on the original CPT alkaloid led to the development of the new analogues irinotecan (CPT-11), topotecan, and 9-aminocamptothecin, which have improved water solubility and lower toxicity. CPT analogues exhibit interesting pharmacokinetic/pharmacodynamic and metabolic properties that are of major research and clinical interest. This review describes the clinical pharmacology of these 3 CPT analogues. Specific areas such as absorption after extravascular administration, pharmacokinetic/pharmacodynamic variability, metabolism, and administration in special populations are discussed. PMID- 9750028 TI - Multiplicity of biliary excretion mechanisms for the camptothecin derivative irinotecan (CPT-11), its metabolite SN-38, and its glucuronide: role of canalicular multispecific organic anion transporter and P-glycoprotein. AB - A frequent dose-limiting effect of irinotecan (CPT-11) is its gastrointestinal toxicity (diarrhea), which is thought to be related to biliary excretion of CPT 11 and its metabolites. Accordingly, we have investigated the mechanism of biliary excretion of these compounds. In vivo pharmacokinetic studies revealed that the biliary excretion of the four anionic forms of CPT-11 and its metabolites was reduced in Eisai hyperbilirubinemic rats, which carry a mutation of the hepatic canalicular multispecific organic anion transporter (cMOAT) gene. The protein encoded by this gene is expressed on the bile canalicular membrane and is responsible for the transport of organic anions into bile. Detailed analysis using isolated liver bile canalicular membrane vesicles to identify transport systems showed that cMOAT is responsible for biliary excretion of the low-affinity component of the carboxylate form of CPT-11 and the high-affinity component of both the lactone and carboxylate forms of SN-38 glucuronide. The carboxylate form of SN-38 is transported by cMOAT alone. Transport of the high affinity component of CPT-11 was inhibited by verapamil and PSC-833, but their effect on the transport of its low-affinity component was minimal. In addition, ATP dependence in the uptake of CPT-11 by membrane vesicles obtained from a P glycoprotein (P-gp)-overexpressing cell line was observed. Thus P-gp may be responsible for transport of the high-affinity component of the carboxylate form of CPT-11. PMID- 9750029 TI - Preclinical approach for identifying drug interactions. AB - Pharmacokinetic drug interactions may be divided into two categories: induction and inhibition of enzymes involved in the metabolism of a drug. The induction and inhibition of such enzymes result in decreases or increases in the blood concentrations of the drug, causing drug effects to be altered. Cytochrome P450 (P450 or CYP) is an enzyme responsible for the metabolism of a wide variety of drugs, including some anticancer agents. If a drug with a high affinity to bind to a specific form of P450 is given to a patient in combination with other drugs mainly metabolized by this enzyme, the former may potentiate the pharmacological actions of the latter by preventing their metabolism and thus increasing their serum concentration. Alternatively, if a drug inhibits or inactivates essentially all forms of P450 nonspecifically, it may be possible that the pharmacological effects of other drugs used in combination with it will be enhanced. CYP3A4 is one of the major forms of P450 in human liver microsomes. In previous studies using human liver microsomes, docetaxel was determined to be metabolized mainly by this isozyme. Thus it was assumed that inducers and inhibitors of CYP3A4 might affect the pharmacokinetics of docetaxel. In our studies, administration of dexamethasone, a known inducer of CYP3A, to mice resulted in decreases in serum docetaxel concentrations. In contrast, ketoconazole, an inhibitor of CYP3A, is assumed to increase the serum and hepatic concentrations of docetaxel. As an example of a drug which might inhibit the metabolism of other drugs, we found that bis-aceto-ammine-dichloro-cyclohexylamine platinum(IV) (JM216), which is currently being developed as a potential anticancer agent, inhibits essentially all major forms of P450 present in human liver microsomes. Since its inhibition potency is relatively high, careful assessment of the effects of this drug on the metabolism of other drugs appears to be necessary. PMID- 9750030 TI - Clinical pharmacology of UCN-01: initial observations and comparison to preclinical models. AB - UCN-01 (7-hydroxystaurosporine; NSC 638850) is a protein kinase antagonist selected for clinical trial based in part on evidence of efficacy in a preclinical renal carcinoma xenograft model. Schedule studies and in vitro studies suggested that a 72-h continuous infusion would be appropriate. In rats and dogs, maximum tolerated doses produced peak plasma concentrations of approximately 0.2-0.3 microM. However, concentrations 10-fold greater are well tolerated in humans, and the compound has a markedly prolonged T1/2. Specific binding to human alpha1-acidic glycoprotein has been demonstrated. These findings reinforce the need to consider actual clinical pharmacology data in "real time" with phase I studies. PMID- 9750031 TI - High-dose chemotherapy and autologous stem cell transplantation as treatment for high-risk breast cancer. AB - High-dose chemotherapy with autologous stem cell transplantation has emerged as a common treatment for patients with breast cancer who have a poor prognosis. The success of this approach appears to depend on the tumor burden and the sensitivity of the disease to chemotherapy because treatment techniques have been refined and treatment-related mortality has declined. Phase II studies in patients with stage II and III disease are encouraging and suggest that treatment with high-dose chemotherapy before the development of metastatic disease may provide an advantage in terms of relapse-free and overall survival. However, tumor cells may contaminate stem cell collections and contribute to relapse after transplantation. Therefore it may be important to separate and select purified CD34+ cells which are not contaminated. It has been suggested that selection bias contributes to the favorable preliminary results observed in phase II studies of high-risk patients. Such issues, together with patient and physician bias regarding the benefits of this strategy, emphasize the need to complete the prospective randomized trials now underway. PMID- 9750032 TI - Phase III randomized comparison of postoperative adjuvant chemotherapy with 2 year oral uracil/tegafur versus 6-cycle cyclophosphamide/methotrexate/5 fluorouracil in high-risk node-negative breast cancer patients. AB - The value of cyclophosphamide/methotrexate/5-fluorouracil (CMF)-type regimens in surgical adjuvant therapy in certain subsets of patients with axillary lymph node negative breast cancer has been evaluated in Europe and the USA. However, Japan has a distinctive standpoint regarding the indications for surgical adjuvant chemotherapy for breast cancer patients. In addition, oral fluoropyrimidines are widely used to treat breast cancer patients in both adjuvant and metastatic settings due to their low toxicity and convenience for long-term administration. Although the antitumor activity and the ability to prolong disease-free survival times of oral fluoropyrimidines have been evaluated in patients with breast cancer, available data are not sufficient to justify replacing CMF-type regimens with oral fluoropyrimidines in postoperative chemotherapy for breast cancer patients. To evaluate the utility of oral fluoropyrimidines in surgical adjuvant chemotherapy, the National Surgical Adjuvant Study Group (N-SAS) was founded in 1995 as a government-funded research group, and nationwide multiinstitutional trials were designed for breast cancer as well as colon and gastric cancers. For high-risk, node-negative breast cancer patients, a prospective randomized trial of surgical adjuvant chemotherapy comparing 6 cycles of CMF with 2 years of daily uracil/tegafur (UFT) started in October 1996. The endpoints of this study include disease-free and overall survival, adverse reactions, quality of life, and cost. PMID- 9750033 TI - Combined-modality treatment for stage IIIa (N2) non-small cell lung cancer: a National Cancer Institute Intergroup study. AB - Induction chemotherapy plus radiotherapy followed by surgery, evaluated in a number of phase II studies, is an effective treatment for patients with stage III A non-small cell lung cancer (NSCLC). The ongoing phase III National Cancer Institute Intergroup study is evaluating concurrent chemotherapy (cisplatin + etoposide)/radiotherapy followed by either surgery or additional radiotherapy followed by 2 additional cycles of chemotherapy in patients with stage IIIA disease. This study will help define the role of surgery and radiotherapy as part of the multimodality therapy of locoregional advanced NSCLC. PMID- 9750034 TI - National Cancer Institute Clinical Trials Program in Colorectal Cancer. AB - Colorectal cancer will be diagnosed in approximately 150,000 patients in the USA this year. Chemotherapy has recently been shown to improve survival when given as adjuvant therapy to surgery in patients with stage III colorectal cancer. Demonstration of this benefit required large, randomized controlled trials. Either 5-fluorouracil (5-FU) and leucovorin for 6 months or 5-FU and levamisole for 12 months are currently considered standard adjuvant treatment for stage III colorectal cancer. However, current adjuvant trials are comparing continuous infusion and intravenous bolus 5-FU regimens and oral uracil/Ftorafur with intravenous 5-FU and leucovorin, as well as studying the timing of chemotherapy in the adjuvant setting. Subsequent adjuvant trials will examine newer regimens with activity in advanced colorectal cancer, as well as the efficacy of monoclonal antibodies. Other trials will study which type of surgery is optimal and whether adjuvant therapy is helpful in stage II colon cancer. Trials in metastatic disease will focus on combinations of newer agents which may improve survival in this patient group. Studies in rectal cancer will focus on determining which agents are optimal in combination with radiation therapy in the adjuvant setting. Molecular characteristics of tumor cells are being defined, which may guide therapy in the future. Careful, logically designed clinical trials will hopefully provide more efficacious therapy for this common cancer. PMID- 9750035 TI - Clinical trials for advanced gastrointestinal cancers in Japan. Japan Clinical Oncology Group Gastrointestinal Oncology Study Group. AB - No standard chemotherapy regimen for the treatment of advanced gastrointestinal cancer exists. However, 5-fluorouracil (5-FU) is being used more frequently as a key component of combination therapy regimens for this disease, and including cisplatin (CDDP) in combination therapy regimens produces significant tumor shrinkage. Although oral formulations of 5-FU are widely used in Japan, their clinical activity and toxicity have not been thoroughly evaluated. In 1992, the Japan Clinical Oncology Group initiated a phase III study in which the survival rates of patients treated with 5-FU 800 mg/m2/day for 5 days by continuous infusion (5-FUci), 5-FUci with CDDP 20 mg/ m2/day for 5 days by infusion, or oral uracil/Ftorafur 375 mg/m2/day with weekly MMC 5 mg/m2 i.v. were compared. This study was closed with 280 patients accrued in 1997, and final analysis was made in early 1998. Irinotecan and paclitaxel are new drugs with activity against gastric cancer. Combination chemotherapy consisting of irinotecan with CDDP has also been evaluated. For treatment of metastatic colorectal cancer, the only active drug previously available in Japan was 5-FU, although 5-FU + leucovorin is the international standard regimen. Irinotecan is now approved in Japan, Europe, and the USA and is the most effective drug for 5-FU-refractory patients. The optimal irinotecan and 5-FU combination regimen is being extensively examined but further trials aimed at accumulating basic data are warranted. PMID- 9750036 TI - The Cooperative Groups: past and future. AB - The Cooperative Group system of the National Cancer Institute (NCI) has been in existence since the 1950s and has evolved to comprise 11 groups, the membership of which includes universities, Community Clinical Oncology Programs, and Cooperative Group Outreach Programs. The Cooperative Groups serve as models for cancer clinical trials throughout the world. However, in today's changing healthcare environment in the USA the Cooperative Groups need to adjust how they operate to ensure the continuation of their leadership role in cancer clinical trials. Government funds, the main source of support for the Cooperative Groups' activities, are shrinking and currently funding is only 50% of the recommended level. If the Cooperative Groups are to remain at the forefront, adjustments must be made in several areas: the Cooperative Groups need to provide an efficient and rapid scientific and legal mechanism to execute large phase III studies of the increasingly important portfolio of compounds being developed by industry more effectively. Industry has come to rely on contract research organizations for expedited testing of their products due to perceived inefficiency in these areas in the Cooperative Group mechanism. The Cooperative Groups are uniquely situated to provide in-depth evaluation of the newest therapies for regulatory agencies and interested health insurers, as well as provide health outcomes data, which are now much sought after by the healthcare industry. Managed care is shaping medical practice, including cancer care, throughout the USA. Finally, the Cooperative Groups need to foster greater international cooperation to speed technology transfers. The leaders of the Cooperative Groups are discussing new approaches to address these deficiencies, while complementing the existing NCI structure and recognizing the independence of each group. The objectives of these new approaches would be: to establish a structure whereby better contracts with industry for conducting trials can be established; to enhance international cooperation in clinical trials; to encourage greater involvement of third-party payers in clinical trials; to build on the scientific breadth of the members; to identify the most appropriate therapies to consider for reimbursement; to establish a framework which builds on the strengths of each of the members; and to integrate health outcomes and economic measures into the protocol activities. The Cooperative Groups are making changes to ensure they remain the leaders in cancer clinical trials well into the 21st century. The benefits of these adjustments will be realized not only by patients, but also by health professionals and the healthcare industry. PMID- 9750038 TI - Cooperative oncology groups in Japan: experience from the Japan Adult Leukemia Study Group. AB - To ensure reliable statistical analysis in clinical trials, a large number of patients is required and therefore multicenter cooperative studies are indispensable in clinical oncology. However, both in the field of oncology and other fields of medicine, well-functioning clinical study groups are rare in Japan. In this review, the reason why multicenter cooperative study groups are difficult to organize in Japan is analyzed. Subsequently, the experience of a self-supporting and successful cooperative study group, the Japan Adult Leukemia Study Group, is reviewed. Finally, in the absence of significant financial support and thus no financial benefit for participating institutions, how to organize a cooperative study group successfully is discussed. PMID- 9750037 TI - Systems of protocol review, quality assurance, and data audit. AB - The US National Cancer Institute (NCI) is the world's largest sponsor of clinical trials in cancer treatment and biology, and it is responsible for the reliability of data generated by means of its funding. The cooperative groups supported by the NCI consist of main academic institutions and smaller affiliates of these institutions. The size of these groups, their geographical dispersion, and the number of studies accruing patients at any one time make it a challenge to ensure that all requirements of institutional oversight, patient consent, protocol compliance, and data submission and quality are met. Each cooperative group has established various procedures for quality assurance. These include data coordinators at the data management center of the group, study chairs, and statisticians. In addition, each group has a committee of physician-investigators and clinical research associates who make periodic site visits to all member institutions to audit the on-site medical records of a sample of patients entered at that institution. The study records are compared with the medical records for all aspects of protocol management and data generation. In addition, adherence to requirements for consent-form signing and oversight by an institutional review board is assessed. Deviations from the study requirements are evaluated as being minor or major. A written report of the audit result is provided to both the NCI and the relevant administrative components of the cooperative group. The audit process has uncovered rare instances of scientific improprieties in these NCI funded clinical trials, but more importantly it has educated investigators and support staff to improve adherence to research and data-collection requirements, which has resulted in greater reliability of study results. PMID- 9750039 TI - European drug development and its impact on national activities: the Dutch example. AB - The development of new anticancer agents is becoming an increasingly complex task which is often beyond the capabilities of a single institution or company. To ensure fast, efficient, high-quality drug development, good coordination and collaboration are essential prerequisites. In this paper, the Dutch national drug development program is described as an example and placed in the perspective of Europe-wide efforts. Since knowledge of new molecular targets and biological approaches by which the malignant growth of cells can be stopped or prevented is increasing, new guidelines for the development of noncytotoxic drugs are required. To avoid duplication and to make the results obtained using these new agents comparable so that patients will gain the maximum benefit of efforts in this field, further extension of international coordination will be of utmost importance in the coming years. PMID- 9750040 TI - Effects of vitamin E and/or C on reactive oxygen species-related lead toxicity in the rat sperm. AB - This study was undertaken to investigate whether treatment with vitamin E (VE) and/or vitamin C (VC) protects rat sperm by inhibiting reactive oxygen species generation induced by lead (Pb) exposure. Male Sprague-Dawley rats were assigned to the following five groups: vitamin-unsupplemented; 150 mg VE/kg chow supplemented; 300 mg VE/kg chow supplemented; 500 mg VC/l drinking water supplemented and 150 mg VE/kg chow + 500 mg VC/l drinking water supplemented group. Rats in each group were divided into Pb-unexposed and Pb-exposed subgroups, received weekly intraperitoneal injection of 10 mg sodium acetate or 10 mg Pb acetate/kg for 6 weeks, respectively. The blood and sperm Pb levels were analyzed by graphite furnace atomic absorption spectrophotometer. Chemiluminescence was measured to evaluate the generation of sperm reactive oxygen species (ROS). Motility and sperm-oocyte penetration rate (SOPR) were measured. In Pb-unexposed rats, epididymal sperm counts, motility, ROS, and SOPR were not different in the five supplemented groups. Lead exposure might decrease the defense capacity of sperm to the oxidative stress and therefore elevate the ROS generation, reduce sperm motility, and reduce SOPR. Supplementation with VE and/or VC reduced ROS generation, prevented loss of motility and capacity of oocyte penetration in Pb-exposed rats. This study suggests that supplementation with VE and/or VC inhibits Pb-related ROS generation, protects spermatozoa from loss of motility and oocyte penetration capability. PMID- 9750041 TI - Antioxidant effects of N-acetylcysteine and succimer in red blood cells from lead exposed rats. AB - This study examined whether lead-induced alterations in selected parameters that are indicative of oxidative stress accompany the toxic effects of lead in red blood cells (RBCs) in vivo. It also explored the possibility that treatment with N-acetylcysteine (NAC) or succimer (meso-2,3-dimercaptosuccinic acid) was capable of reversing parameters indicative of lead-induced oxidative stress. Fisher 344 rats were given 2000 ppm lead acetate in their drinking water for 5 weeks. The lead was then removed and the animals were given NAC (800 mg/kg/day) or succimer (90 mg/kg/day) in their drinking water for 1 week, after which the RBCs were harvested. Animals not given lead and those given lead, but not NAC or succimer, served as negative and positive controls, respectively. At the end of the experiment, blood-lead levels were 35 +/- 4 microg/dl in lead-treated animals, which were reduced to 2.5 +/- 1 microg/dl by treatment with succimer and to 25 +/ 3 microg/dl by treatment with NAC. Lead-exposed animals demonstrated signs of anemia as evidenced by anisocytosis, poikilocytosis, and alterations in hemoglobin, hematocrit, and mean corpuscular volume. Lipid peroxidation, as evidenced by increased malondialdehyde (MDA) content, as well as decreases in reduced glutathione (GSH) and increases in catalase and glucose 6-phosphate dehydrogenase (G6PD) activity were noted in RBCs from lead-treated rats, suggesting that the lead induced oxidative stress. In addition, a significant reduction in blood delta-aminolevulinic acid dehydratase (ALAD) activity suggested that accumulation and autooxidation of delta-aminolevulinic acid might contribute to lead-induced oxidative stress. Treatment with either NAC or succimer reversed lead-induced alterations in MDA and GSH content, but only succimer appeared to partially restore ALAD activity. These results provide in vivo evidence supporting the hypothesis that lead induces oxidative stress in RBCs, which is reversible by treatment with a thiol antioxidant (NAC), as well as a chelating agent (succimer). PMID- 9750042 TI - Effects of lead on rat kidney and liver: GST expression and oxidative stress. AB - The effect of acute exposure to lead acetate on the expression of glutathione S transferase (GST) subunits and the levels of reduced and oxidized glutathione (GSH) and malondialdehyde (MDA) in rat kidney and liver was determined. The purpose of this study was to determine if GSH depletion and/or oxidative stress were responsible for changes in the expression of some or all GSTs that followed lead exposure. In kidney, all GST subunits increased following injection of lead. The level of kidney GSH was not changed at either 0.5 or 1 h after lead exposure, but increased 3, 6, 12 and 24 h after a single injection of lead. MDA levels (a marker of lipid peroxidation) did not change in kidney following lead injection. Immunohistochemical markers of oxidative stress and nitric oxide production were also unchanged by lead administration. Therefore. we conclude that the increases in GST levels in kidney following lead exposure were not dependent on oxidative stress. In liver, lead injection caused GSH depletion (61% of control 12 h after lead treatment) and increased MDA production (2.5-fold increase 6 h after lead exposure), while GSTA1, GSTA2, GSTM1 and GSTM2 did not increase. Analysis of the effects of lead on GST mRNA and GST cellular localization were performed by Northern blot and immunohistochemical techniques. Immunoperoxidase light microscopy and immunogold electron microscopy revealed that the increase in kidney GSTM1 and GSTP1 occurred in nuclei, cytoplasm and microvilli of proximal tubules. Northern blot analysis of GSTA2 and GSTP1 mRNAs showed that their increase following lead exposure was inhibited by actinomycin D, suggesting transcriptional induction. This study demonstrates that acute lead exposure causes dramatic changes in the subcellular distribution and expression of rat kidney GSTs, and that these changes are not a result of oxidative stress. PMID- 9750043 TI - The actions of the H2-blocker cimetidine on the toxicity and biotransformation of the phosphorothioate insecticide parathion. AB - Parathion, like most organophosphorus insecticides currently in use, must undergo cytochrome P450(P450)-dependent activation in order to exert its acute mammalian toxicity (cholinergic crisis). Since P450 isoforms play such an important role in mediating the toxicity of parathion and related insecticides, factors which significantly alter P450 activities, such as exposure to certain xenobiotics, can also be expected to affect the toxicity of these potentially hazardous insecticides. Cimetidine is a H2-histamine antagonist that has been shown to inhibit several P450-isoforms. In addition, administration of cimetidine has been reported to result in clinically significant pharmacokinetic interactions with a wide variety of drugs. In the present study coexposure to cimetidine and parathion resulted in a moderate increase in the toxicity of this pesticide. However, coexposure to cimetidine and paraoxon did not alter the toxicity of the organophosphate, indicating that cimetidine likely affected P450-dependent formation of paraoxon from parathion. In vitro incubations of mouse hepatic microsomes demonstrated that, in addition to reducing the velocity of P450 dependent metabolism of parathion, cimetidine increased the proportion of paraoxon formed (activation). and decreased the proportion of p-nitrophenol formed (detoxification). Since parathion is not eliminated significantly by other routes in the mouse, the bulk of parathion in vivo was metabolized by P450 (although more slowly) in the presence of cimetidine, leading to a greater amount of paraoxon produced, and therefore greater toxicity. Incubations with individual P450 isoforms suggested that cimetidine could act by inhibition of P450 isoforms that detoxify parathion to a greater degree than cimetidine-resistant isoforms, and/or cimetidine could alter the proportions of detoxification versus activation of certain individual isoforms. PMID- 9750044 TI - Human IgE production in hu-PBL-SCID mice injected with birch pollen and diesel exhaust particles. AB - Mice with severe combined immunodeficiency were transplanted with human peripheral blood lymphocytes (hu-PBL-SCID mice). The response to immunisation with birch pollen was used to study possible effects of diesel exhaust particles (DEP) and aluminium hydroxide (Al(OH)3) on human IgE production in this human in vivo model. The adjuvants were well tolerated, as determined by the number of human cells in the peritoneal cavity at the end of the experiments. Total and birch pollen-specific IgE was detected in 76 and 41% of the mice, respectively. In the present experiments where the mice were stimulated early with birch pollen, a doubling in percentage of hu-PBL-SCID mice with production of specific IgE was observed, as compared to later stimulation used in previous experiments. Although a tendency to higher total IgE levels was observed after treatment with DEP, no statistically significant adjuvant effect of DEP or Al(OH)3 could be demonstrated. Electron microscopy analysis after immunogold labelling showed that the major birch pollen allergen Bet v I was released from the pollen grains and adsorbed to the surface of the DEP. Early stimulation with allergen appears to be important for optimal production of specific IgE in the hu-PBL-SCID model. However, our results show that further improvements are needed in order to demonstrate the expected effects from adjuvants and environmental pollutants. PMID- 9750045 TI - [Guidelines for the evaluation of internal quality control of smears for screening of uterine cancer in France in the structures of Pathologic Anatomy and Cytology. French Association for Quality Assurance in Pathologic Anatomy and Cytology (AFAQAP)--Commission for cervical smears]. AB - A national organized mass-screening effective programme is the only way to reduce the risk of cervical cancer, if properly organized and correlated with a system of Quality Assurance. Since 1900, an Association for Quality Assurance was created by the French pathologists, named "AFAQAP". These pathologists thus demonstrated their interest in this kind of action that should be effective if women and clinicians are also implied. The pathologists have concluded the first part of their programme with these French guidelines for internal quality control of pap smears. PMID- 9750046 TI - [Practice guidelines in Pathologic Anatomy and Cytology. Document prepared by the French Association of Quality Assurance in Pathologic Anatomy and Cytology (AFAQAP). Commission No. 4: organization and functioning of Pathologic Anatomy and Cytology]. PMID- 9750047 TI - Change in sexual practice among Australian men who have sex with men, 1992-1996. PMID- 9750048 TI - Society for the Study of Behavioural Phenotypes 7th annual meeting. Cambridge, United Kingdom. November 13-14, 1997. Abstracts. PMID- 9750049 TI - Technical papers composing the proceedings of the 34th annual Rocky Mountain Bioengineering Symposium and the 34th International ISA Biomedical Sciences Instrumentation Symposium. Dayton, Ohio, USA.April 11-13, 1997. PMID- 9750050 TI - Solvent accessibility analysis on the mutants of Hsc70 ATPase fragment. AB - Molecular chaperones are the cellular proteins which mediate the correct folding of other polypeptides. The concept of 'solvent accessibility' is one of the most powerful tools to understand the structure and stability of protein molecules. The hydrophobic variation of amino acid residues due to point mutations at many active sites of chaperone protein Hsc70 using solvent accessibility analysis is carried out. The numerical indices for several properties of amino acid residues, such as, reduction in accessibility, preference of amino acid residues in interior and surface parts, transfer free energy and the preference of amino acid residues to change their positions (buried/exposed) due to amino acid substitutions for Hsc70 and its mutants were set up. The accessibility of amino acid residues varies much between native and mutant proteins whereas there is no major changes on their conformations. The conformational stability for Hsc70 and its mutants were established and the computed hydrophobic free energy change is around 10 kcal/mol due to single amino acid substitution. PMID- 9750052 TI - [A subunit Pasteurella multocida vaccine]. PMID- 9750051 TI - [A case of intestinal amebiasis]. PMID- 9750053 TI - [Eosinophilia in parasitic diseases]. PMID- 9750054 TI - Symptom attenuation by a normally virulent satellite RNA of turnip crinkle virus is associated with the coat protein open reading frame. AB - Many satellite RNAs (sat-RNAs) can attenuate or intensify the symptoms produced by their helper virus. Sat-RNA C, associated with turnip crinkle virus (TCV), was previously found to intensify the symptoms of TCV on all plants in which TCV produced visible symptoms. However, when the coat protein open reading frame (ORF) of TCV was precisely exchanged with that of cardamine chlorotic fleck virus, sat-RNA C attenuated the moderate symptoms of the chimeric virus when Arabidopsis plants were coinoculated with the chimeric virus. Symptom attenuation was correlated with a reduction in viral RNA levels in inoculated and uninoculated leaves. In protoplasts, the presence of sat-RNA C resulted in a reduction of approximately 70% in the chimeric viral genomic RNA at 44 hr postinoculation, whereas the sat-RNA wa consistently amplified to higher levels by the chimeric virus than by wild-type TCV. TCV with a deletion of the coat protein ORF also resulted in a similar increase in sat-RNA C levels in protoplasts, indicating that the TVC coat protein, or its ORF, downregulates the synthesis of sat-RNA C. These results suggest that the coat protein or its ORF is a viral determinant for symptom modulation by sat-RNA C, and symptom attenuation is at least partly due to inhibition of virus accumulation. PMID- 9750055 TI - Plagiarism in the International Journal of Occupational Medicine and Environmental Health (formerly Polish Journal of Occupational Medicine), 1992, 5: 251-256. PMID- 9750056 TI - Accrediting bodies will mesh performance standards. PMID- 9750058 TI - NCQA to count performance measures in accreditation score. PMID- 9750057 TI - Education level, values influence pursuit of alternative therapies. PMID- 9750059 TI - Panic-disorder treatment guidelines published. PMID- 9750060 TI - Clopidogrel, aspirin, and first line therapy. PMID- 9750061 TI - Manifestations of Crohn's disease. PMID- 9750062 TI - Proceedings of the Conference on Early Childhood Caries. Bethesda, Maryland, USA. October 1997. PMID- 9750063 TI - American Association of Electrodiagnostic Medicine 44th annual scientific meeting. San Diego, California, USA. September 19-20, 1997. Abstracts. PMID- 9750064 TI - Proceedings of the 31st Congress of the Japan Epilepsy Society. Kyoto, Japan. September 18-19, 1997. PMID- 9750065 TI - State of the Art in Ultrasonography and Present Day Imaging Modalities of the Breast. Proceedings of the 38th Congress of the Italian Association of Medical Radiology (SIRM). Milan, Italy, May 23-27, 1998. PMID- 9750066 TI - Radiopaque entities of the maxillary sinus. PMID- 9750068 TI - [Normal pressure glaucoma]. PMID- 9750067 TI - It's scary out there. PMID- 9750069 TI - Licensure reform. PMID- 9750071 TI - To CON or not to CON. PMID- 9750070 TI - Cachet, no cash. PMID- 9750073 TI - [The 8th Conference on Valve and Valvular Diseases. Tokyo, Japan, August 2, 1997. Proceedings]. PMID- 9750072 TI - [Molecular biology and molecular genetics in the field of otorhinolaryngology- Special reference to apoptosis]. PMID- 9750074 TI - [Significance of blood levels of E-cadherin for diagnosis of colonic cancer]. PMID- 9750075 TI - [Human tumor sensitivity to 5-fluorouracil influenced by dihydropyrimidine dehydrogenase mRNA in the tumor]. PMID- 9750076 TI - [Percutaneous transesophageal gastric-tube drainage--Development of the balloon catheter and future prospects]. PMID- 9750077 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 29-1998. A 57-year-old man with fever and jaundice after intravesical instillation of bacille Calmette-Guerin for bladder cancer. PMID- 9750078 TI - Alternative therapies for the treatment of childhood cancer. PMID- 9750079 TI - Adulterants in Asian patent medicines. PMID- 9750080 TI - Butyrolactone-induced central nervous system depression after ingestion of RenewTrient, a "dietary supplement". PMID- 9750081 TI - Progressive multifocal leukoencephalopathy, HIV, and highly active antiretroviral therapy. PMID- 9750082 TI - Cytokine therapy in metastatic renal cancer. PMID- 9750083 TI - Cytokine therapy in metastatic renal cancer. PMID- 9750085 TI - Lamotrigine for generalized seizures associated with the Lennox-Gastaut syndrome. PMID- 9750084 TI - Cytokine therapy in metastatic renal cancer. PMID- 9750086 TI - Cervical-disk herniation. PMID- 9750087 TI - Cervical-disk herniation. PMID- 9750088 TI - Ovarian adrenal-like tissue in congenital adrenal hyperplasia. PMID- 9750089 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 30-1998. A 30-year-old woman with increasing hypertension and proteinuria. PMID- 9750090 TI - Multidrug-resistant Salmonella enterica serotype typhimurium infections. PMID- 9750091 TI - Intranasal influenzavirus vaccine in children. PMID- 9750092 TI - Intranasal influenzavirus vaccine in children. PMID- 9750093 TI - Cancer among offspring of survivors of childhood cancer. PMID- 9750094 TI - Secondary acute leukemia in chronic lymphocytic leukemia. PMID- 9750095 TI - Molecular diagnosis of hereditary nonpolyposis colon cancer. PMID- 9750096 TI - Urinary erythrophagocytes in proliferative glomerulonephritis. PMID- 9750097 TI - Treating unrelated disorders in patients with chronic disease. PMID- 9750098 TI - Treating unrelated disorders in patients with chronic disease. PMID- 9750099 TI - Treating unrelated disorders in patients with chronic disease. PMID- 9750100 TI - Treating unrelated disorders in patients with chronic disease. PMID- 9750101 TI - Requests for a physician's help during airline flights. PMID- 9750102 TI - Guns and homicide in the home. PMID- 9750103 TI - The challenge of infectious disease; the European perspective. Proceedings of a symposium. Amsterdam, The Netherlands, 18-19 December 1996. PMID- 9750105 TI - The electronic Plant Gene Register. PMID- 9750104 TI - Photorepair mutants of Arabidopsis. AB - UV radiation induces two major DNA damage products, the cyclobutane pyrimidine dimer (CPD) and, at a lower frequency, the pyrimidine (6-4) pyrimidinone dimer (6 4 product). Although Escherichia coli and Saccharomyes cerevisiae produce a CPD specific photolyase that eliminates only this class of dimer, Arabidopsis thaliana, Drosphila melanogaster, Crotalus atrox, and Xenopus laevis have recently been shown to photoreactivate both CPDs and 6-4 products. We describe the isolation and characterization of two new classes of mutants of Arabidopsis, termed uvr2 and uvr3, that are defective in the photoreactivation of CPDs and 6-4 products, respectively. We demonstrate that the CPD photolyase mutation is genetically linked to a DNA sequence encoding a type II (metazoan) CPD photolyase. In addition, we are able to generate plants in which only CPDs or 6-4 products are photoreactivated in the nuclear genome by exposing these mutants to UV light and then allowing them to repair one or the other class of dimers. This provides us with a unique opportunity to study the biological consequences of each of these two major UV-induced photoproducts in an intact living system. PMID- 9750106 TI - HIV prevention with drug using populations. Current status and future prospects. PMID- 9750107 TI - Britain hunts down CJD epidemic in removed appendixes. PMID- 9750108 TI - New HIV strain could pose health threat. PMID- 9750110 TI - Breathalyzer device sniffs for disease. PMID- 9750109 TI - New timepiece has a familiar ring. PMID- 9750111 TI - Which of our genes makes us human? PMID- 9750112 TI - How a growth control path takes a wrong turn to cancer. PMID- 9750113 TI - The rightness of copyright. PMID- 9750114 TI - Errors in genome reviews. PMID- 9750115 TI - Who should own scientific papers? PMID- 9750117 TI - Duplicating a tangled genome. PMID- 9750116 TI - Driving the growth cone. PMID- 9750118 TI - Proceedings of the 2nd Seminar on Food-Borne Parasitic Zoonoses: Current Problems, Epidemiology, Food Safety and Control. Khon Kaen, Thailand, 6-9 December 1995. PMID- 9750119 TI - Proceedings of the final meeting of Concerted Action CT93 0909. Lelystad, 28-29 April 1997. PMID- 9750120 TI - [Dissertations defended in 1998]. PMID- 9750121 TI - ABO announces 1999 Ketcham Award recipients--Charles J. Burstone and Dale B. Wade. PMID- 9750122 TI - PZ271: a microsatellite showing polymorphism in cattle. PMID- 9750123 TI - Strain IMB-1, a novel bacterium for the removal of methyl bromide in fumigated agricultural soils. AB - A facultatively methylotrophic bacterium, strain IMB-1, that has been isolated from agricultural soil grows on methyl bromide (MeBr), methyl iodide, methyl chloride, and methylated amines, as well as on glucose, pyruvate, or acetate. Phylogenetic analysis of its 16S rRNA gene sequence indicates that strain IMB-1 classes in the alpha subgroup of the class Proteobacteria and is closely related to members of the genus Rhizobium. The ability of strain IMB-1 to oxidize MeBr to CO2 is constitutive in cells regardless of the growth substrate. Addition of cell suspensions of strain IMB-1 to soils greatly accelerates the oxidation of MeBr, as does pretreatment of soils with low concentrations of methyl iodide. These results suggest that soil treatment strategies can be devised whereby bacteria can effectively consume MeBr during field fumigations, which would diminish or eliminate the outward flux of MeBr to the atmosphere. PMID- 9750124 TI - Coupling capillary high-performance liquid chromatography to matrix-assisted laser desorption/ionization mass spectrometry and N-terminal sequencing of peptides via automated microblotting onto membrane substrates. AB - To minimize low-quantity sample handling for protein sequencing and matrix assisted laser desorption/ionization (MALDI) mass spectrometry, a system consisting of an HPLC interfaced to an automated blotting device was used for off line sample collection. Typically, protein digests are separated by reverse-phase HPLC and the resulting peptide fractions are pooled, concentrated, and then subjected to N-terminal sequence analysis. Obtaining unambiguous sequence from peptides derived from protein digestion at subpicomole levels requires careful sample handling to prevent loss of sample. In cases where multiple sequences are present, a secondary method such as mass spectrometry is needed to confirm the identity of the peptides. To minimize sample handling, commercial microblotting instruments have become available to deposit peptides directly onto polyvinylidene difluoride (PVDF) membrane for automated N-terminal sequence analysis. In order to adapt this technology to mass spectrometry, we investigated the use of MALDI-MS compatible membranes such as Teflon and polyethylene (PE) as the blotting media for fraction collection. Using a panel of standard peptides as well as protein digests, we demonstrate that peptides separated by capillary HPLC can be collected directly onto Teflon or PE and detected into the femtomole range. Furthermore, detailed sequence analysis could be obtained by postsource decay fragmentation spectra of individual peptides blotted onto either PE or Teflon. Due to the high sensitivity of the MALDI-MS from these membranes, it was discovered that the small amount of peptide that passed through the PVDF membrane during a collection of peptides for N-terminal sequencing was sufficient to be collected and mass analyzed from a second underlying MALDI-MS compatible membrane. Therefore, from a single HPLC separation, samples could be collected onto both PVDF for traditional N-terminal sequencing and PE or Teflon for MALDI MS. We demonstrate the general utility of this method for sequencing peptides from a tryptic digestion at subpicomole levels and for identifying unknown proteins separated by 2-dimensional gel electrophoresis. The ability to generate both N-terminal sequence and confirmatory mass information from multiple peptides in a single separation greatly improves the reliability and the accuracy of protein characterization at subpicomole levels. PMID- 9750125 TI - Identification of biotinylation sites on proteins by selective retrieval of 2 iminobiotinylated peptides from proteolytic peptide mixtures: localization of the accessible lysine residues on the photosystem I subunits PsaD and PsaE. AB - An affinity purification technique was established that allows the selective isolation of 2-iminobiotinylated peptides from proteolytic digest of proteins in order to identify surface-exposed protein domains. Serving as model systems, two photosystem I subunits, PsaD and PsaE from the cyanobacterium Synechococcus elongatus, were overexpressed in Escherichia coli, modified in vitrowith NHS-2 iminobiotin which incorporates 2-iminobiotin at exposed amino groups, and subjected to proteolytic digestion by Glu-C and Arg-C protease, respectively. 2 Iminobiotin-containing proteolytic peptides were subsequently extracted from the proteolytic digests using avidin agarose in a batch procedure and the extracted peptides were separated by HPLC chromatography. The analysis of the peptide maps by electrospray ionization mass spectrometry or N-terminal sequencing showed that avidin-extracted peptide fractions contain almost exclusively 2-iminobiotinylated proteolytic fragments of PsaE or PsaD. No unmodified peptides of PsaD or PsaE were detected. According to this analysis, PsaE is accessible to biotinylation at all of its 7 lysine residues and at its N-terminus. Similarly, all 11 lysine residues of PsaD can be biotinylated and only the N-terminus of PsaD is not accessible. PMID- 9750126 TI - Site-specific, enzymatic biotinylation of recombinant proteins in Spodoptera frugiperda cells using biotin acceptor peptides. AB - Site-specific, enzymatic biotinylation of recombinant proteins can be exploited to circumvent many problems associated with the use of biotinylating reagents in vitro and to overcome some of their inherent limitations. Additionally, biotinyl proteins can be purified to near-homogeneity in a single step under native conditions. Here we report that a biotin acceptor peptide (BAP) substrate for Escherichia coli biotin holoenzyme synthetase (BirA) can be used to label recombinant proteins with biotin in Spodoptera frugiperda (Sf9) cells, and we describe a collection of baculovirus transfer vectors specifically designed for this purpose. These BioBac vectors will greatly expand the range of proteins to which this technology can be applied. PMID- 9750127 TI - A spectrophotometric method for the determination of proteins damaged by oxidized lipids. AB - The Ehrlich reaction was optimized to determine pyrrolized proteins produced as a consequence of lipid peroxidation and oxidative stress. The procedure consisted of the treatment of the modified protein with p-(dimethylamino)benzaldehyde at a controlled acidity and temperature, and the determination of adducts produced against the blank obtained in the absence of the reagent. The extinction coefficient of Ehrlich adducts was calculated by using epsilon-N pyrrolylnorleucine (Pnl) as standard and was 35,000 M-1 cm-1. The response was linear and reproducible within the range 0. 16-20 microM Pnl. The assay was applied to determination of pyrrole content in bovine serum albumin, bovine alpha globulins, bovine gamma-globulins, and mixtures of them, incubated overnight with 1 mM of 4,5(E)-epoxy-2(E)-heptenal, obtaining results similar to those from determination of Pnl by capillary electrophoresis after basic hydrolysis of the protein. The method was also applied to pyrrole determination in bovine plasma proteins either incubated with epoxyalkenals, hydroxyalkenals, lipid hydroperoxides, and secondary products of lipid peroxidation, or oxidized with Fe3+/ascorbate. All these treatments produced pyrrolization of plasma proteins and all Ehrlich adducts gave very similar absorbance spectra with the exception of that produced in the treatment with hydroxyalkenals. The above results suggest that protein pyrrolization is a normal consequence of the lipid peroxidation process and of oxidative stress, and that Ehrlich adducts may be valid to determine this pyrrolization. PMID- 9750128 TI - Light-scattering submicroscopic particles as highly fluorescent analogs and their use as tracer labels in clinical and biological applications. AB - Submicroscopic gold particle suspensions scatter colored light when illuminated with white light, and we have observed that a light-scattering gold particle suspension has the same appearance as a fluorescing solution. Thus, when illuminated by a narrow beam of white light, a 40-nm gold sol displays a clear (not cloudy), green scattered light (Tyndall) beam and has the same appearance as a fluorescing fluorescein solution. These, as well as other, observations have suggested to us that, in general, light-scattering particles can be treated as fluorescent analogs and used as fluorescent analog tracers in immuno- and DNA probe assays as well as in cell and molecular biology studies. Light-scattering particles are advantageous in these applications because particles such as gold and silver have very high light-scattering powers, which allows these particles to be easily detected, by light-scattering, at particle concentrations as low as 10(-16) M. The scattered light can be detected by the unaided eye for qualitative measurements or with a simple light-sensitive detector for quantitative measurements. Moreover, individual particles can be easily detected by eye or a video camera using a simple light microscope with a proper illuminating system. In addition, submicroscopic particles which scatter blue, green, yellow, orange, or red light can be readily synthesized. Antibodies, DNA probes, and other tracer substances can be readily attached to gold and other particles without altering their light-scattering properties. In this article we present the theory which allows one to predict the light-scattering properties of particles of different sizes and compositions and identify those particle sizes and compositions which appear most adequate for particular applications. Furthermore, we calculate molar extinction coefficients and emission efficiencies for particles of different sizes and compositions which allows us to compare the light-producing powers of these particles with those of well known fluorescent tracers. A 60-nm gold particle, for example, is equivalent to about 3 x 10(5) fluorescein molecules. Very simple, easy to use, low-cost, ultrasensitive immuno- and DNA probe assays can be developed using light-scattering particles as fluorescent analog tracers. Single particles can be detected on cell surfaces and inside cells using light microscopy techniques with proper illumination as described in the article. At high particle densities, particle-labeled cells have the same appearance as fluorescent cells. PMID- 9750129 TI - Light-scattering submicroscopic particles as highly fluorescent analogs and their use as tracer labels in clinical and biological applications. AB - In a companion article we present the idea that submicroscopic light-scattering particles, such as gold and silver particles, can be used as fluorescent analog tracers in biological and clinical applications. The light-emitting power and scattered light color of the particles can be adjusted by changing particle composition or diameter. Using Rayleigh and Mie light-scattering theory, we calculated the absorbance and light-scattering properties and power of particles of different diameters and selected compositions and compared them to the properties of fluorophores. In the present article, we evaluate experimentally the optical properties of particles of selected compositions and sizes and compare the results with the theoretically calculated values. We also discuss the methods which we use to measure the light-scattering properties of particles in suspension and to view individual particles by light microscopy. We also outline examples which demonstrate the use of light-scattering particles as fluorescent analogs in biological and clinical applications. PMID- 9750130 TI - Quantitative analysis of modified antisense oligonucleotides in biological fluids using cationic nanoparticles for solid-phase extraction. AB - Based on a novel method for solid-phase extraction using cationic polystyrene nanoparticles, the suitability of the extraction procedure for quantitation of terminally and backbone-modified antisense oligonucleotides was investigated. Extractions were carried out from both human plasma and urine. Quantitative analysis of the extracted samples was performed with capillary gel electrophoresis. In accordance with previous results obtained with phosphorothioate oligonucleotides in human plasma, high linearity and accuracy of the assay was demonstrated for an oligodeoxyribonucleotide-palmityl conjugate as well as for a modified oligoribonucleotide. Optimized extraction conditions allow the isolation of oligonucleotides in high yields and purity even for concentrations in the low nanomolar range, down to 5 nM. Comparing the results obtained from human plasma and urine, no significant differences in the absolute recovery rates which reach values up to 95% were observed. However, when the loading capacity of the nanoparticles was exceeded, selective recovery was observed for the coisolation of phosphodiester and phosphorothioate oligonucleotides. This effect can be explained by differences in the attractive forces between PO- and PS-oligonucleotides and the particle surface and appears to be valuable for a modification-dependent enrichment of oligonucleotides out of complex mixtures. PMID- 9750131 TI - Dot far-western blot analysis of relative binding affinities of the Src homology 3 domains of Efs and its related proteins. AB - The Src homology 3 (SH3) domains are a modular structure of about 60 amino acid residues found in many proteins important in signal transduction. Each SH3 domain has a binding specificity to sequences containing a PXXP motif in ligand proteins. We found that a focal adhesion kinase (FAK)-related protein, cell adhesion kinase beta (CAKbeta), was bound in vitro by the SH3 domain of embryonal Fyn-associated substrate (Efs), a docking protein structurally related to p130Cas (Cas) and HEF1. Here, we employed a dot far-Western blotting technique to evaluate the affinity and specificity of the binding by the SH3 domains of Efs and its related proteins. The SH3 domains and their ligands were prepared as glutathione S-transferase fusion proteins, and one of the binding components was immobilized on membranes while the other was labeled with 32P to use as a probe. The amount of the bound probe was determined by autoradiography using an imaging plate and a bioimaging analyzer. A competitive binding assay showed that Efs, compared with Cas and HEF1, had a SH3 domain with a lower relative affinity to CAKbeta and FAK and with a preference to interact with FAK rather than CAKbeta. Our assay based on dot far-Western blotting is a simple and sensitive method to evaluate fine differences in the binding affinity of SH3-mediated interactions. PMID- 9750132 TI - Comparison of fluorescent genotyping methods. PMID- 9750133 TI - Optimization of derivatization with 2-aminobenzoic acid for determination of monosaccharide composition by capillary electrophoresis. PMID- 9750134 TI - Efficient carbohydrate release, purification, and derivatization. PMID- 9750135 TI - Second-order kinetic analysis of IAsys biosensor data: its use and applicability. AB - The kinetic analysis of IAsys biosensor association data usually relies upon the assumption of constant ligate concentration. In certain circumstances this assumption may no longer be valid. In a similar vein, the analysis of the dissociation phase assumes the concentration of ligate to be negligible in the liquid phase-an assumption that may not be sustainable for high-affinity interactions. In this paper we derive analytical solutions of the second-order differential kinetic equations for the association and dissociation phases, together with a binding isotherm that also allows for changes in concentration of both the ligand and the ligate. Using these equations it is possible to determine the conditions under which the pseudo-first-order assumption ceases to be valid. It is found that the effect of ligate depletion on the association rate constant becomes significant only when binding low ligate concentrations to ligand on surfaces with high binding capacities or high affinities. Similarly, the rebinding in the dissociation phase is dependent upon the affinity and the ligand capacity together with the starting dissociation response compared to the capacity. Finally, depletion also affects the form of the binding isotherm, particularly in situations involving high matrix capacities for ligate and high affinity interactions. PMID- 9750136 TI - Measurement of hydrogen peroxide in biological samples containing high levels of ascorbic acid. AB - The physiological concentration of hydrogen peroxide in the aqueous humor was reported to range between 25 and 60 microM, and conditions leading to elevated levels could have important damaging effects such as cataract formation. However, the high concentration of ascorbic acid in aqueous humor, which is 20 times that of plasma, was recently shown to interfere in the dichlorophenol-indophenol assay for hydrogen peroxide. The actual concentration of hydrogen peroxide in this fluid has become a controversial issue. In the present study, we used the method of ferrous oxidation of xylenol orange (FOX1 assay) performed in a nitrogen atmosphere to accurately measure low levels of hydrogen peroxide, even in the presence of ascorbic acid at concentrations normally found in aqueous humor. Contrary to values reported in the literature, we observed that the concentration of hydrogen peroxide in the rabbit aqueous humor is less than 5 microM, which is the detection limit of the method. PMID- 9750137 TI - Simultaneous analysis of the majority of low-molecular-weight, redox-active compounds from mitochondria. AB - Studies of the interaction between oxidative stress and mitochondrial dysfunction are complicated by analytical limitations, especially the need to assess multiple parameters in relatively small samples. We have addressed this problem by developing a methodology for the simultaneous analysis of the majority of low molecular-weight, redox-active compounds from mitochondria using HPLC separations followed by coulometric array detection. The method described should also be applicable for the study of redox-active compounds in other subcellular organelles as well as in intact cells and tissues. The protocol described enables simultaneous measurement of antioxidants (e.g., tocopherols, ascorbate, lipoates, uric acid, and glutathione), markers of oxidative stress (e.g., o-tyrosine, m tyrosine, nitrotyrosine, dityrosine, glutathione disulfide, and 8 hydroxydeoxyguanosine) as well as other metabolites (e.g., purines and indoles). In all, ca. 600 redox active compounds can be detected, most with a limit of detection of approximately 5 pg on column. Results, including analytical parameters, from a study of liver mitochondria from control and diabetic rats are presented to demonstrate utility of this methodology. PMID- 9750138 TI - Zero-length protein-nucleic acid crosslinking by radical-generating coordination complexes as a probe for analysis of protein-DNA interactions in vitro and in vivo. AB - Redox-active coordination complexes such as 1,10-phenanthroline-Cu(II) (OP-Cu) and bleomycin-Fe(III) are commonly used as "chemical nucleases" to introduce single-strand breaks in nucleic acids. Here we report that under certain conditions these complexes may crosslink proteins to nucleic acids. In vitro experiments suggest that proteins are crosslinked to DNA by a mechanism similar to dimethyl sulfate-induced crosslinking. Furthermore, we demonstrate that the OP Cu complex can generate protein-DNA crosslinks in mammalian cells in vivo. By combining the OP-Cu crosslinking and a "protein shadow" hybridization assay we identify proteins interacting with DNA in isolated pea chloroplasts and show that this methodology can be applied to detect DNA-binding proteins on specific DNA sequences either in vitro or in vivo. PMID- 9750139 TI - A double stain for total and oxidized proteins from two-dimensional fingerprints. AB - Oxidative modification of proteins plays a major role in the etiology of aging and age-related diseases. For example, in Alzheimer's disease, although evidence points to oxidation of proteins as a causative factor in loss of cognitive abilities, it is not known which specific proteins of the brain are most susceptible to these modifications. Thus, it is of interest to identify the specific proteins which are susceptible to oxidation in vivo. Two-dimensional protein fingerprint methods offer the analytical potential for resolution of thousands of individual proteins from tissues, and the oxidized proteins can be visualized with immunological probes. Sensitive methods permit recovery and sufficient amino acid sequencing to identify these proteins. However, for such analyses it is essential to simultaneously analyze both protein content and level of oxidation. We have evaluated several approaches, identified the sources of artifacts and interferences, and developed a double-staining procedure that allows visualization and quantitation of total protein patterns as well as the specific oxidized proteins from two-dimensional protein fingerprints. The method has been applied to cells grown in culture and to tissue extracts from young and old animals. PMID- 9750140 TI - Measurement of hepatic glucose output, krebs cycle, and gluconeogenic fluxes by NMR analysis of a single plasma glucose sample. AB - 13C and 1H NMR spectroscopy of plasma glucose was used to resolve the isotopomer contributions from tracer levels of [1,6-13C2]glucose, a novel tracer of glucose carbon skeleton turnover, and [U-13C]propionate, a tracer of hepatic citric acid cycle metabolism. This allowed simultaneous measurements of hepatic glucose production and citric acid cycle fluxes from the NMR analysis of a single plasma glucose sample in fasted animals. Glucose carbon skeleton turnover, as reported by the dilution of [1,6-13C2]glucose, was 56 +/- 2 micromol/kg/min in the presence of labeling from [U-13C]propionate and 53 +/- 4 micromol/kg/min in its absence. Therefore, as expected, the labeling contributions from [U 13C]propionate metabolism did not have a significant effect on the measurement of glucose turnover. For the group infused with both tracers, citric acid cycle flux estimates from the analysis of glucose C2 isotopomer ratios were consistent with those from our recent experiments where only [U-13C]propionate was infused, verifying that the presence of [1,6-13C2]glucose did not interfere with these measurements. This integrated analysis of hepatic glucose output and citric acid cycle fluxes from plasma glucose isotopomers yielded a noninvasive estimate of hepatic citrate synthase flux of 74 +/- 12 micromol/kg/min for 24-h fasted rats. PMID- 9750141 TI - A mathematical model describing tannin-protein association. AB - A quantitative model is derived for tannin-protein binding and protein precipitation in solution. This model is based on the assumption that precipitation occurs when the number of tannin molecules associated with one protein molecule attains a critical value. Precipitation occurs at this point because tannin crosslinking causes formation of large protein aggregates. Analytical expressions were derived for the dependence of protein precipitate yields upon the concentrations of the protein and tannin in solution. This expression fits reasonably well the experimentally observed bell-shaped dependencies of bovine serum albumin or gelatin precipitation upon total protein and tannin concentrations. PMID- 9750142 TI - A continuous fluorimetric assay for tail-specific protease. AB - A continuous fluorimetric assay for tail-specific protease (Tsp) has been developed using a fluorescence donor/quencher system, in which 5-[(2-aminoethyl) amino]naphthalene-1-sulfonic acid (EDANS) and 4-(4-dimethylaminophenylazo)benzoic acid (DABCYL) are attached to the N-terminus and the lysyl side chain of peptide AARAAK-(6-aminocaproyl)2-ENYALAA, respectively. Tsp-mediated cleavage of the Ala Arg peptide bond separates the quencher, DABCYL, from the donor, EDANS, and results in a large increase in the fluorescent yield of EDANS (>50-fold). Using this sensitive assay, Escherichia coli tail-specific protease was shown to exhibit typical Michaelis-Menten kinetics with a kcat of 0.086 +/- 0.002 s-1, KM of 4.0 +/- 0.3 microM, and kcat/KM of 2.2 x 10(4) M-1 s-1. A control substrate, which only differs from the above substrate by having a charged residue (glutamate) at the C-terminus, showed drastically reduced activity to Tsp (kcat/KM = 58 M-1 s-1). A peptide containing the C-terminal sequence of the substrate, GRGYALAA, was shown to be a competitive inhibitor of Tsp with a KI value of 31 microM. These results demonstrate the utility of this assay for the rapid assessment of Tsp activity. PMID- 9750143 TI - A homogeneous chemiluminescent assay for telomerase. AB - I have designed an unamplified assay for human telomerase based on the hybridization protection assay. This assay measures in vitro synthesis of the human telomeric repeat DNA sequence (TTAGGG)N by solution hybridization with a chemiluminescent acridinium ester oligonucleotide probe. The assay is capable of detecting less than 0. 1 fmol of the telomerase repeat DNA sequence, and has a linear range extending to 3.5 fmol/reaction. Using this assay, telomerase activity can be measured in less than 60 min. The hybridization protection assay can be used to determine the amount of telomerase activity in a cell-free extract obtained from as few as 10(5) cells, and is three to four orders of magnitude less sensitive than PCR-based telomerase assays. However, the precision, accuracy, and rapidity of this telomerase assay make it suitable for enzyme isolation, enzyme characterization, and high-throughput screening applications. PMID- 9750144 TI - Use of the yeast three-hybrid system as a tool to study caspases. AB - Caspases are a family of heteromeric (p20/p10) cysteine proteases with important functions in the regulation of apoptosis and inflammation. Up to now, tools to identify new substrates for caspases have mostly been limited to the random screening of in vitro translated proteins that are known, or assumed, to play a role in apoptosis. We describe the use of a yeast three-hybrid approach as a tool that adapts the classical two-hybrid system to the needs of heteromeric caspases for functional dissection of known interactions or screening for physiological substrates and inhibitors. Functional heteromeric caspase-1 was obtained by coexpression of p20(Cys285Ser) and p10 caspase-1 subunits that were each fused to the Gal4 DNA-binding domain. Upon coexpression of a third hybrid of the Gal4 activation domain and the viral caspase-1 pseudosubstrate inhibitors CrmA or p35, or the prototype physiological caspase-1 substrate prointerleukin-1beta, a functional Gal4 transcription factor could be reconstituted. In contrast, no interaction was found between CrmA or p35 and the immature p45 or p30 precursor forms of caspase-1. Therefore, the three-hybrid system might allow screening for new physiological substrates and inhibitors of heteromeric caspases. PMID- 9750145 TI - Identification of oxidized proteins based on sodium dodecyl sulfate polyacrylamide gel electrophoresis, immunochemical detection, isoelectric focusing, and microsequencing. AB - It has been demonstrated that an age-associated increase in protein oxidation is a highly selective rather than random phenomenon. Addition of carbonyl groups is a widely used marker for the detection of oxidative damage to the proteins. Carbonyls can be detected immunochemically using antibodies against dinitrophenylhydrazine (DNPH), a reagent that specifically reacts with protein carbonyl groups. In this paper, a procedure for the identification of oxidized proteins during aging or under pathological conditions is described. Protein samples were treated with DNPH and then separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), followed by immunochemical detection. Proteins exhibiting positive immunostain were then purified by SDS PAGE followed by isoelectric focusing. The focused protein band was further concentrated by another run of SDS-PAGE and transferred onto Immobilon-P membrane. The identity of the protein was revealed by automated Edman microsequencing and a computer database search. This method is successfully demonstrated for the identification of housefly arginine kinase, a cytosolic protein that is involved in the energy metabolism of insect muscle cells. PMID- 9750146 TI - A robust method for determining DNA binding constants using capillary zone electrophoresis. AB - Capillary zone electrophoresis (CZE or CE) with on-line UV detection was utilized to measure the binding constants between purified calf thymus DNA and a library of designed tetrapeptides which had been constructed using unnatural amino acids with thiazole ring side chains. Mixtures containing a constant amount of a tetrapeptide, the neutral marker (mesityl oxide), and varying concentrations of DNA were prepared and equilibrated at 8 degreesC for 12 h. CE was then utilized to separate unbound tetrapeptides from the DNA-peptide complex. The UV absorbance of the peak representing unbound tetrapeptide decreased incrementally as a result of increasing the concentration of DNA in the equilibrium mixture. The absorbance of the peak corresponding to the unbound tetrapeptide was obtained directly from the electropherogram and used in the calculation of the DNA-peptide binding constants. The binding constant for each tetrapeptide to calf thymus DNA was obtained from the negative slope of a Scatchard plot and a comparison of the binding constants for different peptides showed that the tetrapeptides in the library have DNA-binding affinities ranging from 10(2) to 10(6) M-1. PMID- 9750147 TI - Differential silver-staining sodium dodecyl sulfate-polyacrylamide gel electrophoresis: a nonisotopic method for characterizing gel-separated histone DNA complexes. AB - Some nonspecific, DNA-binding proteins, like the linker histones, precipitate DNA upon binding. This is a poorly understood process that limits analysis of such nucleoprotein complexes using standard gel electrophoresis. To circumvent this problem, low concentrations of glutaraldehyde were used to crosslink the linker histones to DNA; then the partially crosslinked complexes were solubilized in SDS2 and separated by SDS-PAGE. Differential detection was accomplished using two different silver staining protocols that preferentially stained either proteins or nucleic acids. A technique was developed which allows the relative proportion of linker histones and DNAs in each detected band to be determined, and is referred to as differential staining SDS-PAGE (DS-SDS-PAGE). DS-SDS-PAGE provides a novel, non-isotopic means for characterizing multiple nucleoprotein bands separated by polyacrylamide gel electrophoresis. In applying this method to a model linker histone-DNA study, we were able to detect both protein-DNA and protein-protein contacts that are important in linker histone assembly onto DNA. PMID- 9750148 TI - Degradable dUMP outer primers in merged tandem (M/T)-nested PCR: low- and single copy DNA target amplification. AB - PCR amplification of DNA from a single initiating genomic molecule or low-copy template often requires two sequential amplification reactions with nested primer pairs to achieve the necessary specificity and sensitivity. Residual outer primers can result in undesired primer activity during the inner nested cycles. To circumvent this problem, we have used dU-containing primers for first round amplification and then uracil N-glycosylase (UNG) to degrade them and the ends of their dU-primer-containing amplified DNA products. We have applied this method to the detection of an exon 11 mutation in the HEXA gene. We have merged the step of a single-tube PCR amplification with outer dU primers with a tandem amplification using non-dU-nested primers (hence, the term merged tandem-nested or M/T-nested PCR). Serial dilutions of genomic DNA showed that this method could amplify a specific target from as few as three haploid genome equivalents of template DNA. Specific products were obtained from the DNA of single cells in 19 of 20 replicates, using 12 outer and 28 inner nested PCR cycles, with an intervening UNG digestion step. When coupled with heteroduplex mutational analysis, this method reliably distinguished mutant versus wild-type HEXA gene fragments amplified from single cells without primer artifact. PMID- 9750149 TI - Protein identification at the low femtomole level from silver-stained gels using a new fritless electrospray interface for liquid chromatography-microspray and nanospray mass spectrometry. AB - Conventional capillary liquid chromatography/mass spectrometry (LC/MS) typically employs low microl/min flow rates with gas/liquid sheath to enhance spray stability. Over the past several years a number of reports have demonstrated success with electrospray (ES) interface designs optimized for submicroliter/min flows which have clear advantages in terms of enhancement of detection limit, lower sample consumption, and ability to accommodate a wider range of buffer conditions. We report here a fritless electrospray interface (FESI) design that is inexpensive and robust and can be operated and adapted to accommodate a variety of applications for submicroliter/min flow rates. The novelty of this interface revolves around the use of a fritless microcapillary column and precolumn application of electrospray voltage at a microtee junction to achieve stable microspray and nanospray flow rates. This sheathless FESI device eliminates postcolumn dead volume since small particles (/= 8.0. The yield of 4-nitrosophenol continues to increase even after pH 11.1, 1. 2 units above the pKa of phenol, suggesting that the phenolate ion, and not phenol, is involved in the reaction. Hydrogen peroxide is not formed as a by-product. The nitrosation reaction is zero-order in phenol and first-order in peroxynitrite, suggesting the phenolate ion reacts with an activated nitrosating species derived from peroxynitrite, and not with peroxynitrite itself. Under optimal conditions, the yields of 4-nitrosophenol are comparable to those of 2- and 4-nitrophenols, indicating that the nitrosation reaction is as significant as the nitration of phenolic compounds by peroxynitrite. Low concentrations of CO2 facilitate the nitrosation reaction, but excess CO2 dramatically reduces the yield of 4-nitrosophenol. The dual effects of CO2 can be rationalized if O=N-OO- reacts with the peroxynitrite anion-CO2 adduct (O=N-OOCO-2) or secondary intermediates derived from it, including the nitrocarbonate anion (O2N-OCO-2), the carbonate radical (CO*-3), and *NO2. The product resulting from these reactions can be envisioned as an activated intermediate X-N=O (where X is -OONO2, -NO2, or -CO-3) that could transfer a nitrosyl cation (NO+) to the phenolate ion. An alternative mechanism for the nitrosation of phenol involves the one-electron oxidation of the phenolate ion by CO*-3 to give the phenoxyl radical and the oxidation of O=N-OO- by CO*-3 to give a nitrosyldioxyl radical (O=N-OO*), which decomposes to give *NO and O2; the *NO then reacts with the phenoxyl radical giving nitrosophenol. Both mechanisms are consistent with the high yields of NO-2 and O2 during the alkaline decomposition of peroxynitrite and the potent inhibitory effect of N-3 on the nitrosation of phenol by peroxynitrite and peroxynitrite/CO2 adducts. The biological significance of the peroxynitrite-mediated nitrosations is discussed. PMID- 9750161 TI - Calculation of standard transformed formation properties of biochemical reactants and standard apparent reduction potentials of half reactions. AB - The standard Gibbs energies of formation and standard enthalpies of formation of species involved in biochemical reactions are used to calculate standard transformed Gibbs energies of formation and standard transformed enthalpies of formation of 62 biochemical reactants (sums of species) at 298.15 K, pH 7, and ionic strengths of 0, 0.10, and 0.25 M. It has been possible to put the oxidized and reduced forms of some reactants in this table because their standard apparent reduction potentials are known at pH 7. This paper emphasizes redox reactions. Two applications have been made of these 62 new values of standard transformed Gibbs energies of formation at pH 7: (1) They have been used to calculate standard transformed Gibbs energies of formation of 16 more biochemical reactants from measurements of apparent equilibrium constants of redox reactions. (2) They have been used to calculate standard apparent reduction potentials at pH 7 for half reactions involving reactants discussed in this article and the previous one. This table of standard apparent reduction potentials can be extended considerably from known apparent equilibrium constants for enzyme-catalyzed redox reactions. This brings the total number of reactants for which the standard transformed Gibbs energy of formation at 298K, pH 7, and ionic strengths of 0, 0.10, and 0.25 M have been calculated to 142. PMID- 9750162 TI - Differential enantioselectivity of murine glutathione S-transferase isoenzymes in the glutathione conjugation of trans-3,4-dihydroxy-1, 2-oxy-1,2,3,4 tetrahydrobenzo[c]phenanthrene stereoisomers. AB - The kinetics of the glutathione (GSH) conjugation of (+)- and (-)-enantiomers of anti- as well as syn-3,4-dihydroxy-1,2-oxy-1,2,3, 4 tetrahydrobenzo[c]phenanthrene (B[c]PDE) catalyzed by murine GSH S-transferase (GST) isoenzymes has been investigated. Murine GSTs exhibited significant differences in their enantioselectivity toward B[c]PDE stereoisomers. For example, while pi class isoenzyme mGSTP1-1 was virtually inactive toward stereoisomers with 1S configuration [(-)-syn-and (+)-anti-B[c]PDE], these stereoisomers were good substrates for alpha class isoenzyme mGSTA1-2. When GST activity was measured as a function of varying B[c]PDE concentration (10-320 microM) at a fixed saturating concentration of GSH (2 mM), each isoenzyme examined obeyed Michaelis-Menten kinetics with all four B[c]PDE stereoisomers. Alpha class isoenzyme mGSTA4-4 exhibited negligible activity toward all four stereoisomers of B[c]PDE. The catalytic efficiency of mGSTA1-2 was approximately 1.5- to 15-fold higher than other murine GSTs in the GSH conjugation of (-)-anti B[c]PDE, which among the four B[c]PDE stereoisomers is the most potent pulmonary carcinogen in the newborn mouse model and a potent skin tumor-initiator. While alpha class isoenzymes mGSTA3-3 and mGSTA1-2 were equally efficient in the GSH conjugation of (+)-anti-B[c]PDE, their catalytic efficiencies toward this stereoisomer were significantly higher than those of mGSTP1-1 and mGSTM1-1. Likewise, mGSTA1-2 was relatively more efficient than other GSTs in the GSH conjugation of both enantiomers of syn-B[c]PDE. In summary, our results indicate that (a) murine GSTs significantly differ in their enantioselectivity in the GSH conjugation of B[c]PDE stereoisomers, which may partially account for the observed differences in the carcinogenic potency of B[c]PDE stereoisomers, and (b) mGSTA1-2 and mGSTA3-3 play a major role in the detoxification of B[c]PDE. PMID- 9750163 TI - The roles of threonine-136 and glutamate-137 of human medium chain acyl-CoA dehydrogenase in FAD binding and peptide folding using site-directed mutagenesis: creation of an FAD-dependent mutant, T136D. AB - We studied the roles of Thr-136 (T136) and Glu-137 (E137) in the biogenesis of medium chain acyl-CoA dehydrogenase (MCAD) by altering the former to Ser (T136S), Asp (T136D), or Leu (T136L) and the latter to Asp (E137D), Gln (E137Q), or Lys (E137K). After import into mitochondria, T136S and E137D were assembled into the native tetramer as efficiently as the wild-type. The tetrameric assembly of four other variants with a nonconservative substitution was severely impaired. When expressed in Escherichia coli as the mature subunit, the amounts of the catalytically active forms of T136S and E137D were comparable to wild-type, whereas four nonconservative variants were lost as aggregates. Of these nonconservative variants, only T136D formed catalytically active tetramer when the culture broth and buffers were supplemented with riboflavin and FAD, respectively. Culturing T136L or E137K at a lower temperature (28 degreesC) did not increase the yield at all, suggesting the severity of disruption of biogenesis. These results, together with the previous crystallographic findings, indicate that the T136 hydroxyl is a major FAD-binding site, and that E137 carboxyl plays a key role in the beta-domain folding, through salt bridge formation with K164. These findings also support the notion that the isoalloxazine ring plays a critical role in the MCAD folding, presumably exerting nucleating effects. PMID- 9750164 TI - Synergistic induction by collagen and fibronectin of liver-specific genes in rat primary cultured hepatocytes. AB - The extracellular matrix plays an important role for maintaining liver functions. We examined the effects of type I collagen and fibronectin on the expression of liver-specific genes in rat primary hepatocytes. When primary culture hepatocytes were overlaid with a type I collagen-gel, the expression of liver-specific genes (tyrosine aminotransferase, aldolase B, and albumin) increased by 4-5 times, compared with not overlaid hepatocytes. In contrast, the expression of non-liver specific genes (GAPDH and beta-actin) was suppressed under the same conditions. The addition of fibronectin together with type I collagen-gel further enhanced the expression of liver-specific genes by 1.4-1.8 times. The addition of GRGDS peptide instead of fibronectin with the collagen-gel had a similar effect on hepatic gene expression to that of fibronectin. Addition of fibronectin alone exhibited had no effect on gene expression. These results suggest that type I collagen and fibronectin synergistically induce liver-specific genes. PMID- 9750165 TI - Identification of the uridine diphosphate glucuronosyltransferase isoform UGT1A6 in rat brain and in primary cultures of neurons and astrocytes. AB - The expression of a phenol uridine diphosphate glucuronosyltransferase (UGT) was investigated in rat brain homogenate and in primary cultures of astrocytes and neurons, by means of model substrates (1-naphthol and 4-methylumbelliferone) assays, Western blot analysis and reverse transcription-polymerase chain reaction (RT-PCR) experiments. Glucuronidation of these substances occurred in cerebral cell or brain homogenates, although to different extents. The specific activity was the highest in astrocytes, with values more than 10- and 100-fold those found in neurons or total brain, respectively. Using antibodies able to recognize several rat liver UGT isoforms, only one protein with an apparent molecular mass of 54 kDa was detected in astrocyte and neuron homogenates and brain microsomes. RT-PCR experiments run with primers specifically designed for the rat liver UGT1A6 revealed amplificons of the expected sizes in accordance with the presence of UGT1A6 mRNA. The nucleotide sequence of the 330-base pair product was 100% homologous to that of exon 1 of rat liver isoform UGT1A6. In conclusion, this work allowed us to identify for the first time a constitutive cerebral UGT isoform identical to rat liver UGT1A6, which glucuronidates planar phenolic substances in cultured astrocytes, neurons, and the entire brain. PMID- 9750166 TI - Neuronal nitric oxide synthase-membrane phospholipid interactions. AB - Most of the neuronal nitric oxide synthase (nNOS) is present in the particulate fraction of tissue extracts. Here, we show that the calmodulin (CaM)-binding domain of nNOS interacts with anionic phospholipid vesicles but not with neutral ones. Identification of residues in the CaM-binding domain of nNOS as the key domain for the interaction is also documented. Recombinant wild-type nNOS was found to associate with phosphatidylserine (PS) or phosphatidic acid (PA) but not with phosphatidylethanolamine (PE) or phosphatidylcholine (PC), indicating that nNOS-phospholipid binding requires an electrostatic interaction. A synthetic peptide corresponding to residues 732-754 blocked the interaction of nNOS with PS. Furthermore, a purified fusion protein containing residues 724-755 interacted with PS in a competitive fashion with CaM. Inactive nNOS lacking CaM-binding ability, generated by mutation of (Lys732LysLeu) to (Asp732AspGlu) (Watanabe, Y., Hu, Y., and Hidaka, H., FEBS Lett. 403, 75-78, 1997) did not interact with PS. Preincubation of nNOS with PS protected subsequent limited proteolysis of the synthase by Staphylococcus aureus V8 protease, probably as a result of conformational changes in the protein. Wild-type nNOS was found almost entirely in the membrane fraction of Sf9 cells, whereas inactive nNOS was also found in cytosolic fraction in Sf9 cells expressing the mutant enzyme. These results demonstrate that the mutated hydrophobic/basic amino acid cluster in nNOS sequence, Lys732LysLeu, is essential for nNOS-PS and nNOS-CaM interactions. PMID- 9750167 TI - Nitric oxide-dependent induction of glutathione synthesis through increased expression of gamma-glutamylcysteine synthetase. AB - The nitric oxide (NO) donors S-nitrosopenicillamine or DetaNONOate, which release NO at a rate of 0-15 nM sec-1, were exposed to rat aortic vascular smooth muscle cells for a period of 0-24 h. This treatment resulted in an increase in total glutathione levels of two- to threefold under conditions where no cytotoxicity was detected. The signaling pathways do not involve activation of protein kinase G Ialpha nor are they cGMP dependent. Oxidation of reduced glutathione (GSH) was found after exposure to NO for 3-4 h at rates of formation at or above 8 nM sec 1. Increased intracellular GSH was due to enhanced expression of the rate limiting enzyme for GSH synthesis, gamma-glutamylcysteine synthetase. Since NO has been shown previously to protect cells against oxidative stress, we propose that the increase in GSH by NO is a potential mechanism for enhancing the antioxidant defenses of the cell. This result also has important implications for identifying redox-sensitive cell signaling pathways that can be activated by NO. PMID- 9750168 TI - A CT repeat in the promoter of the chicken malic enzyme gene is essential for function at an alternative transcription start site. AB - CT repeats are abundant in eukaryotic genomes and have been implicated in a number of biological events. The promoter of the chicken malic enzyme gene contains a long polypyrimidine/polypurine tract that includes seven tandem CTs. This CT repeat region together with 14 immediately downstream nucleotides functions as an active alternative promoter when linked to a reporter gene and may direct transcription initiation at a cluster of minor sites in the endogenous gene [G. Xu and A. G. Goodridge (1996) J. Biol. Chem. 271, 16008-16019]. In the sequence required for promoter activity, -105 to -83 bp, there are two purines; only the A at -83 bp influences promoter activity. Mutation of different four nucleotide stretches of the CT repeats to purines decreased promoter activity as a function of the increase in GC content. Increasing the number of CT repeats by changing pyrimidines downstream of (CT)7 to CTs increased promoter activity. These sequences and other regions showed moderate sensitivity to S1 nuclease in supercoiled plasmids, suggesting the presence of non-B-DNA structures. Increasing the length of the CT repeats should increase the propensity to adopt non-B-DNA structures such as triplexes. Constructs with 10, 15, or 22 repeats had increased expression relative to wild type. Thus, the ability of CT repeats to form non-B DNA structures may be functionally important. PMID- 9750169 TI - Modulation of human flavin-containing monooxygenase 3 activity by tricyclic antidepressants and other agents: importance of residue 428. AB - Human flavin-containing monooxygenase 3 (FMO3) is subject to modulation by tricyclic antidepressants and other agents. Imipramine activates FMO3-catalyzed metabolism of methimazole at all substrate concentrations tested. This distinguishes FMO3 from rabbit FMO1 and FMO2, which are activated at high substrate concentration and inhibited at low substrate concentration, and pig FMO1, which is inhibited at all substrate concentrations. The response of FMO3 is also unique in that chlorpromazine is markedly more effective as a modulator than is imipramine. n-Octylamine, MgCl2, and HgCl2 all inhibit FMO3, the first two in a biphasic manner. Substitution of lysine for threonine at position 428 significantly alters the response of FMO3 to modulators without changing the kinetic parameters for the metabolism of the substrate. Activation by imipramine and chlorpromazine is reduced or abolished and inhibition, most obvious at low substrate concentrations, is observed. This is consistent with elimination of self-activation in the metabolism of imipramine. The mutation at 428 also eliminates the biphasic nature of the inhibition by n-octylamine and MgCl2, but does not alter the effect of HgCl2. Our findings show that the activity of FMO3 can be modulated by large drug molecules as well as short-chain amines and metal ions. This modulation can be markedly altered by changing a single amino acid in the enzyme. PMID- 9750170 TI - Transcriptional inhibitory role of the tail domains of histone (H3 x H4)2 tetramers. AB - Histone-DNA templates for bacteriophage T7 RNA polymerase were assembled from a plasmid containing a promoter and a terminator for this polymerase, (H3 x H4)2 tetramers deprived of their tail domains, and H2A x H2B dimers. Histone (H3 x H4)2 tetramers lacking their terminal domains were obtained from trypsin-digested nucleosomal cores. The oligonucleosomal templates containing (H3 x H4)2 tetramers lacking their tail domains, like the control templates with intact core histone octamers, protect approximately 146 base pairs of DNA against micrococcal nuclease digestion. The transcriptional inhibition caused by the association of DNA with core histone octamers is significantly reduced upon elimination of the tail domains of the (H3 x H4)2 tetramers. Apparently, the terminal domains of (H3 x H4)2 must be present to block transcription efficiently. These results show the important inhibitory role played by the tail domains of the histone (H3 x H4)2 tetramers, suggesting the involvement of these regions in transcriptional regulation. PMID- 9750171 TI - Engineering of pyridine nucleotide specificity of nitrate reductase: mutagenesis of recombinant cytochrome b reductase fragment of Neurospora crassa NADPH:Nitrate reductase. AB - The cytochrome b reductase fragment of Neurospora crassa NADPH:nitrate reductase (EC 1.6.6.3) was overexpressed in Escherichia coli with a His-tag for purification after mutation of the NADPH binding site. The recombinant enzyme fragment was altered by site-directed mutagenesis guided by the three-dimensional structure of cytochrome b reductase fragment of corn NADH:nitrate reductase (EC 1.6.6.1). Substitution of Asp for Ser920 (using residue numbering for holo NADPH:nitrate reductase of N. crassa) greatly increased preference for NADH. This mutant had nearly the same NADH:ferricyanide reductase kcat as wild-type with NADPH. Substitutions for Arg921 had little influence on coenzyme specificity, while substitution of Ser or Gln for Arg932 did. The cytochrome b reductase mutant with greatest preference for NADH over NADPH was the doubly substituted form, Asp for Ser920/Ser for Arg932, but it had low activity and low affinity for coenzymes, which indicated a general loss of specificity in the binding site. Steady-state kinetic constants were determined for wild type and mutants with NADPH and NADH. Wild type had a specificity ratio of 1100, which was defined as the catalytic efficiency (kcat/Km) for NADPH divided by catalytic efficiency for NADH, while Asp for Ser920 mutant had a ratio of 0.17. Thus, the specificity ratio was reversed by over 6000-fold by a single mutation. Preference for NADPH versus NADH is strongly influenced by presence/absence of a negatively charged amino acid side chain in the binding site for the 2' phosphate of NADPH in nitrate reductase, which may partially account for existence of bispecific NAD(P)H:nitrate reductases (EC 1.6.6.2). PMID- 9750172 TI - Functional role of cysteine residues in the Na+/H+ exchanger effects of mutation of cysteine residues on targeting and activity of the Na+/H+ exchanger. AB - We investigated the role of cysteine residues in activity and localization of the NHE1 isoform of the Na+/H+ exchanger. Each of the nine cysteine residues was mutated to serine or arginine. Mutation of the first serine (amino acid number 9) and serine number six (amino acid number 477) resulted in dramatic decreases in detectable activity of the Na+/H+ exchanger when transfected into AP-1 cells. Some other mutations resulted in minor decreases in activity of the protein. Confocal and light microscopy of mutant cells with decreased activity showed that the antiporter protein was mostly retained in an intracellular compartment which colocalized with the medial-Golgi cisternae. Smaller amounts of active protein still remained targeted to the plasma membrane in these mutants. Treatment of wild-type cells with DTT also caused the retention of the Na+/H+ exchanger to the same intracellular compartment. The results suggest that cysteines play an important role in intracellular folding and trafficking of the Na+/H+ exchanger. PMID- 9750173 TI - A drug-responsive and protease-resistant peripheral NADH oxidase complex from the surface of HeLa S cells. AB - Our laboratory described a ca. 34-kDa protein of the HeLa S cell surface that bound an antitumor sulfonylurea N-(4-methylphenylsulfonyl)-N'-(4-chlorophenyl) urea (LY181984) with high affinity and that exhibited NADH oxidase and protein disulfide-thiol interchange activities also inhibited by LY181984. The quinone site inhibitor 8-methyl-N-vanillyl-6-noneamide (capsaicin) also blocked these same enzymatic activities. Using capsaicin inhibition as the criterion, the drug responsive oxidase was released from the surface of HeLa S cells and purified. The activity of the released capsaicin-inhibited oxidase was resistant to heating at 50 degrees C and to protease digestion. After heating and proteinase K digestion, the activity was isolated in >90% yield by FPLC as an apparent 50- to 60-kDa multimer. Final purification by preparative SDS-PAGE yielded a capsaicin inhibited NADH oxidase activity of a specific activity indicative of >500-fold purification relative to the plasma membrane. The final activity correlated with a ca. 34-kDa band on SDS-PAGE. Matrix-assisted laser desorption mass spectroscopy as well as reelectrophoresis of the 34-kDa band indicated that the ca. 34-kDa material was a stable mixture of 22-, 17-, and 9.5-kDa components which occasionally migrated as a ca. 52-kDa complex. The purified complex tended to multimerize and formed insoluble 10- to 20-nm-diameter amyloid rods. The components of the purified 34-kDa complex were blocked to N-terminal amino acid sequencing and were resistant to further protease digestion. After multimerization into amyloid rods, the protein remained resistant to proteases even under denaturing conditions and to cyanogen bromide either with or without prior alkylation. PMID- 9750174 TI - Prolyl endopeptidase from Sphingomonas capsulata: isolation and characterization of the enzyme and nucleotide sequence of the gene. AB - Prolyl endopeptidase (prolyl oligopeptidase, EC 3.4.21.26) was purified from Sphingomonas capsulata IFO 12533, and its gene was cloned and expressed in Escherichia coli. The recombinant enzyme was markedly inhibited by diisopropyl phosphofluoridate and hardly affected by SH reagents or metal chelators, similar to the native enzyme purified from S. capsulata. Nucleotide sequencing analysis revealed an open reading frame of 2169 bp, coding for a protein of 723 amino acids with a predicted molecular weight of 78,433. The amino acid sequence was 39.6, 45.3, 38.9, and 38.3% homologous to Flavobacterium meningosepticum, Aeromonas hydrophila, porcine brain, and human T cell prolyl endopeptidase, respectively. A region near the C-terminus and the region containing the putative catalytic triad residues were highly conserved. The enzyme was crystallized by the hanging drop vapor diffusion method, using ammonium sulfate as a precipitant. PMID- 9750175 TI - DNA binding activity of the aryl hydrocarbon receptor is sensitive to redox changes in intact cells. AB - The potential involvement of vicinal dithiols in the transformation of the aryl hydrocarbon (Ah) receptor from its ligand binding to DNA binding form in Hepa-1 cells was explored through the use of diamide and phenylarsine oxide (PAO), which have been shown to specifically form a stable ring complex with vicinal sulfhydryl groups in selected proteins. Pretreatment with diamide and PAO rapidly prevented the inducer-dependent formation of the Ah receptor/xenobiotic response element complex detected by electrophoretic mobility shift assays and suppressed Ah receptor-mediated transcription. Diamide and PAO also inhibited DNA binding activity of the nuclear Ah receptor subsequent to its translocation to the nucleus but to a lesser extent than that observed with pretreatment conditions. The Ah receptor exhibited much higher sensitivity to cellular redox changes than Sp1, a transcription factor previously shown to be very sensitive to redox regulation. Diamide added to nuclear extracts inhibited Ah receptor DNA binding more than when it was added in intact cells. In contrast, Ah receptor DNA binding activity was more sensitive to PAO when it was added to intact cells than when it was added to nuclear extracts. Finally, dithiol 2,3-dimercaptopropanol was over 100 times more effective than monothiol 2-mercaptoethanol in reversing the PAO dependent inhibition of Ah receptor DNA binding activity. This suggests that vicinal sulfhydryl residues may be involved in DNA binding of the Ah receptor. PMID- 9750176 TI - Triphenyltin as an inhibitor of membrane-bound pyrophosphatase of Rhodospirillum rubrum. AB - The effect of triphenyltin on the activity of membrane-bound pyrophosphatase of Rhodospirillum rubrum was investigated. Triphenyltin inhibits the hydrolysis of chromatophore membrane-bound pyrophosphatase in a pH-dependent pattern, being maximal at pH 9-10. At basic pH values, the inhibition produced by this organotin on membrane-bound pyrophosphatase is very similar to that produced on the chromatophore H+ATPase (I50 = 14.4 and 10 microM, respectively). Detergent solubilized membrane-bound pyrophosphatase is also inhibited by triphenyltin, but the cytoplasmic enzyme of R. rubrum is inhibited only slightly. The inhibitory effect of triphenyltin on membrane-bound pyrophosphatase is the same with Mg-PPi or Zn-PPi, and is dependent on the chromatophore membrane concentration. Triphenyltin modified mainly the Vmax of the enzyme, and only slightly its Km. Free Mg2+ does not reverse the inhibition. Reducing agents prevent triphenyltin inhibition of the membrane-bound pyrophosphatase, but their effect is due to an alteration of the inhibitor, and not to a modification of thiol groups of the enzyme. The most likely site for triphenyltin inhibition in chromatophore membrane-bound pyrophosphatase is a component either within or closely associated with the membrane. PMID- 9750177 TI - Disulfide isomerization within the C-terminus of cobrotoxin decelerates by thiol compounds and trinitrophenylation, but accelerates by modification of carboxyl groups. AB - A disulfide isomerization at the C-terminus of cobrotoxin occurred spontaneously by dissolving in alkali buffer. Irreversible conversion of cobrotoxin into its isomers was completely achieved within 4 days. The isomerization reaction was decelerated by thiol compounds including GSSG, GSH, cystamine, and cysteamine in a pseudo-first-order kinetic, and GSSG was the most effective one among the thiol compounds used. Moreover, the oxidized thiol compounds were always superior to reduced ones in decreasing the rate of disulfide interchange. To further assess the intrinsic elements essential for the occurrence of disulfide isomerization of cobrotoxin, the toxin molecule was subjected to modification on its Arg, Lys, Trp, Tyr, and carboxyl groups. In sharp contrast to other modified derivatives, the isomerization reaction was decelerated by trinitrophenylation on Lys-26, Lys 27, and Lys-47, whereas it was rapidly completed after modification of carboxyl groups. Neither chemical modification nor the toxin's conformation affected the irreversibility of isomerization reaction. Thus, the observed change in the rate of disulfide isomerization reflects the involvement of Lys residues and carboxyl groups in this reaction. Although thiol compounds further decelerated the conversion of trinitrophenylated cobrotoxin into its isomers, they did not exert a notable effect on the isomerization of carboxyl groups-modified derivative. These results clearly indicate that disulfide isomerization of cobrotoxin is, in part, driven by the positively charged Lys residues at positions 26, 27, and 47 of the toxin molecule, and that the thiol compounds are coordinated with the negatively charged groups of cobrotoxin to exert their inhibitory action. PMID- 9750178 TI - Effects of drying methods and additives on structure and function of actin: mechanisms of dehydration-induced damage and its inhibition. AB - Limited stability impedes the development of industrial and pharmaceutical proteins. Dried formulations are theoretically more stable, but the drying process itself causes structural damage leading to loss of activity after rehydration. Lyophilization is the most common method used to dry proteins, but involves freezing and dehydration, which are both damaging to protein. We compared an air-drying method to freeze-drying to test the hypothesis that terminal dehydration is the critical stress leading to loss of activity. The secondary structure of air-dried and freeze-dried actin was analyzed by infrared spectroscopy and related to the level of activity recovered from the rehydrated samples. Actin dried by either method in the absence of stabilizers was highly unfolded and the capacity to polymerize was lost upon rehydration. The degree of unfolding was reduced by air-drying or freeze-drying actin with sucrose, and the level of activity recovered upon rehydration increased. The addition of dextran to sucrose improved the recovery of activity from freeze-dried, but not air-dried samples. Dextran alone failed to protect the structure and function of actin dried by either method, indicating that proteins are not protected from dehydration-induced damage by formation of a glassy matrix. In some cases, recovered activity did not correlate directly with the level of structural protection conferred by a particular additive. This result suggests that secondary structural protection during drying is a necessary but not sufficient condition for the recovery of activity from a dried protein after rehydration. PMID- 9750180 TI - Characterization of the promoter region of the rat neprilysin gene. AB - The neprilysin gene is composed of three distinct 5' noncoding exons which can be joined to the first coding exon to generate multiple mRNA species, all encoding the same protein. Genomic fragments containing upstream sequences of each of these three noncoding exons from the rat neprilysin gene were subcloned in the promoterless vector pXp1, which contains the luciferase reporter gene. Expression was compared between a neprilysin positive human spinal cord cell line, HSC-2, and neprilysin negative lines MCF-7, a human breast adenocarcinoma cell line and Hep G2, a human liver carcinoma cell line. The first and second promoter regions showed high activity in the positive cell line, but low activity in the negative cell lines. An analysis of the exon 1 promoter region showed that the proximal 85 nucleotides exhibited basal promoter activity. An enhancer-like sequence was found to be located within a 22-bp fragment located at -136 to -115. Scanning mutagenesis of a 29-bp fragment containing the enhancer-like sequence showed that changes in each 5- or 6-bp segment throughout this fragment decreased activity; however, mutations of the segment encompassing positions 19 to 24 eliminated >98% of the promoter activity. Binding of nuclear proteins from HSC-2 cells to this 29 bp fragment was observed by gel shift analysis. The ability of mutations within the 29-bp fragment to affect enhancer activity correlated with the ability of these mutant oligonucleotides to compete for the wild-type sequence in gel shift assays. PMID- 9750179 TI - The N-terminal region is important for the allosteric activation and inhibition of the Escherichia coli ADP-glucose pyrophosphorylase. AB - The ADPglucose pyrophosphorylase (EC 2.7.7.27) from Escherichia coli is allosterically activated by fructose 1,6-bisphosphate and inhibited by AMP. Proteolysis of the enzyme with proteinase K causes loss of activity and generates two peptides, 21 and 28 kDa, from the 49.7-kDa subunit. The presence of ADPglucose, Mg2+, and fructose 1, 6-bisphosphate during the incubation with proteinase K protected the enzyme activity and prevented cleavage at sites Met181 Ala182 and Phe192-Val193. Proteolysis of the protected enzyme removed 10 to 13 amino acids from the N-terminal and 2 amino acids from the C-terminal. The resulting enzyme was almost independent of the need for fructose 1,6-bisphosphate for maximal activity and insensitive to inhibition by AMP. The apparent affinity for the substrates was similar to that of the fully-activated wild-type enzyme. These data suggest that amino acid residues in the N-terminal portion and possibly the C-terminal portion of ADPglucose pyrophosphorylase are part of the regulatory domain of the enzyme, critical for allosteric regulation of the enzyme. PMID- 9750181 TI - The Unit of Selection in Viscous Populations and the Evolution of Altruism. AB - Group selection can overcome individual selection for selfishness and favour altruism if there is variation among the founders of the spatially distinct groups, and groups with many altruists become substantially larger (or exist longer) than groups with few. Whether altruism can evolve in populations that do not have an alternation of local population growth and global dispersal ("viscous populations") has been disputed for some time. Limited dispersal protects the altruists from the non-altruists, but also hinders the export of altruism. In this article, we used the Pair Approximation technique (tracking the dynamics of pairs of neighbours instead of single individuals) to derive explicit invasion conditions for rare mutants in populations with limited dispersal. In such viscous populations, invading mutants form clusters, and ultimately, invasion conditions depend on the properties of such clusters. Thus there is selection on a higher level than that of the individual; in fact, invasion conditions define the unit of selection in viscous populations. We treat the evolution of altruism as a specific example, but the method is of more general interest. In particular, an important advantage is that the spatial aspects can be incorporated into game theory in a straightforward fashion; we will specify the ESS for a more general model. The invasion condition can be interpreted in terms of inclusive fitness. In contrast with Hamilton's model, the coefficient of relatedness is not merely a given genetical constant but depends on local population dynamical processes (birth, dispersal and death of individuals). With a simple birth rate function, Hamilton's rule is recovered: the cost to the donor should be less than the benefit to the recipient weighted with the coefficient of relatedness. As the coefficient of relatedness is roughly proportional to an individual's number of neighbours, benefits to the recipient must be substantial to outweigh the costs, confirming earlier studies. We discuss the consequences for the evolution of dispersal and outline how the method may be extended to study evolution in interacting populations.Copyright 1998 Academic Press PMID- 9750182 TI - Metabolic Control Analysis: Separable Matrices and Interdependence of Control Coefficients. AB - A central quantity for the analysis of the interdependence of control coefficients is the Jacobian H of the pathway. For a simple metabolic chain, H is known to be tridiagonal. Its inverse H-1, which is required to calculate control coefficients, is semi-separable. A semi-separable nxn matrix (aij) has the characteristic property that it is decomposable into two triangles for each of which there are vectors r=(r1, . . . ,rn) and t=(t1, . . . ,tn) with aij=ritj. The exact definitions of semi-separability and the related separability of matrices are given in Appendix B. Owing to the semi-separability of H-1, the determinants of all 2x2 sub-matrices of elements located within one of the triangles are zero. Therefore, these triangles are regions of vanishing two minors. The flux control coefficient matrix CJ is hown to be separable and the concentration control coefficient matrix Cs to be semi separable. Cs has, in addition, the peculiarity that the row vector is the same for both its upper and lower triangle. A feedback loop gives rise to a new sub-region of vanishing two minors, thereby disturbing the semi-separability of the upper triangle of Cs. A recipe is given to graphically construct the regions of vanishing two-minors of concentration control coefficients. The notion of (semi-)separability allows assessment of all dependences of control coefficients for metabolic pathways.Copyright 1998 Academic Press PMID- 9750183 TI - The Fixed Sample Search Rule and Use of an Indicator Character to Evaluate Mate Quality. AB - The phenotypic correlation observed between mated males and females is a direct result of the decision rules that individuals employ during search for a mate. We generalise the fixed sample search rule and examine how the functional relationship between a male indicator character and the fitness benefit of a mating decision influences female search behaviour. If the phenotypes of males and females combine additively to influence the benefit of a mating decision and fitness is a highly inclined function of phenotypic values the optimal value of n, the number of males that females sample during search, is relatively large. The phenotype of the searcher has no impact on the optimal value of n if fitness is a linear function; the optimal n increases with the phenotype of females if the fitness function is convex; and a concave fitness function induces females with high trait values to sample relatively few males. Because females that sample many potential mates are expected to encounter a male with a high expression of the indicator trait linear, convex and concave fitness functions induce a random, assortive and disassortative combination of phenotypes between mated individuals, respectively. Thus, the fixed sample research tactic can produce the variety of phenotypic correlations observed between mated individuals in natural systems and our ability to derive rule-specific predictions of search behavior may generally require information on how characters used to evaluate potential mates are related to fitness.Copyright 1998 Academic Press PMID- 9750184 TI - Independent Changes of ATP/ADP or DeltapH could cause Oscillations in Photosynthesis. PMID- 9750186 TI - Elucidating the genetic networks of development: a bioinformatics approach. PMID- 9750187 TI - WebWise: guide to the joint genome institute web site. PMID- 9750188 TI - The synuclein family. AB - The synuclein gene family recently came into the spotlight, when one of its members, alpha-synuclein, was found to be mutated in several families with autosomal dominant Parkinson's disease (PD). A peptide of the alpha-synuclein protein had been characterized previously as a major component of amyloid plaques in brains of patients with Alzheimer's disease (AD). The mechanism by which this presynaptic protein is involved in the two most common neurodegenerative disorders, AD and PD, remains unclear. Remarkably, another member of this gene family, gamma-synuclein, has been shown to be overexpressed in breast carcinomas and may also be overexpressed in ovarian cancer. The possible involvement of the synuclein proteins in the etiology of common human diseases has raised exciting questions and is the subject of intense investigation. Details of the properties of any member of the synuclein family may provide useful information for understanding the characteristics and function of other family members. The present review offers a synopsis of the current state of knowledge of all synuclein family members in different species. PMID- 9750189 TI - Imprinting mechanisms. AB - A number of recent studies have provided new insights into mechanisms that regulate genomic imprinting in the mammalian genome. Regions of allele-specific differential methylation (DMRs) are present in all imprinted genes examined. Differential methylation is erased in germ cells at an early stage of their development, and germ-line-specific methylation imprints in DMRs are reestablished around the time of birth. After fertilization, differential methylation is retained in core DMRs despite genome-wide demethylation and de novo methylation during preimplantation and early postimplantation stages. Direct repeats near CG-rich DMRs may be involved in the establishment and maintenance of allele-specific methylation patterns. Imprinted genes tend to be clustered; one important component of clustering is enhancer competition, whereby promoters of linked imprinted genes compete for access to enhancers. Regional organization and spreading of the epigenotype during development is also important and depends on DMRs and imprinting centers. The mechanism of cis spreading of DNA methylation is not known, but precedent is provided by the Xist RNA, which results in X chromosome inactivation in cis. Reading of the somatic imprints could be carried out by transcription factors that are sensitive to methylation, or by methyl cytosine-binding proteins that are involved in transcriptional repression through chromatin remodeling. PMID- 9750190 TI - Comparative gene mapping: a fine-scale survey of chromosome rearrangements between ruminants and humans. AB - A total of 202 genes were cytogenetically mapped to goat chromosomes, multiplying by five the total number of regional gene localizations in domestic ruminants (255). This map encompasses 249 and 173 common anchor loci regularly spaced along human and murine chromosomes, respectively, which makes it possible to perform a genome-wide comparison between three mammalian orders. Twice as many rearrangements as revealed by ZOO-FISH were observed. The average size of conserved fragments could be estimated at 27 and 8 cM with humans and mice, respectively. The position of evolutionary breakpoints often correspond with human chromosome sites known to be vulnerable to rearrangement in neoplasia. Furthermore, 75 microsatellite markers, 30 of which were isolated from gene containing bacterial artificial chromosomes (BACs), were added to the previous goat genetic map, achieving 88% genome coverage. Finally, 124 microsatellites were cytogenetically mapped, which made it possible to physically anchor and orient all the linkage groups. We believe that this comprehensive map will speed up positional cloning projects in domestic ruminants and clarify some aspects of mammalian chromosomal evolution. PMID- 9750191 TI - Compositional heterogeneity within, and uniformity between, DNA sequences of yeast chromosomes. AB - The heterogeneity within, and similarities between, yeast chromosomes are studied. For the former, we show by the size distribution of domains, coding density, size distribution of open reading frames, spatial power spectra, and deviation from binomial distribution for C + G% in large moving windows that there is a strong deviation of the yeast sequences from random sequences. For the latter, not only do we graphically illustrate the similarity for the above mentioned statistics, but we also carry out a rigorous analysis of variance (ANOVA) test. The hypothesis that all yeast chromosomes are similar cannot be rejected by this test. We examine the two possible explanations of this interchromosomal uniformity: a common origin, such as genome-wide duplication (polyploidization), and a concerted evolutionary process. PMID- 9750192 TI - Duplication of a genomic region containing the Cdc2L1-2 and MMP21-22 genes on human chromosome 1p36.3 and their linkage to D1Z2. AB - Cdc2L1 and Cdc2L2 span approximately 140 kb on human chromosome 1p36.3. The products of the Cdc2L genes encode almost identical protein kinases, the PITSLRE kinases, which have functions that may be relevant to the regulation of transcription/splicing and apoptotic signaling. These genes are deleted/translocated in neuroblastomas with MYCN gene amplification, a subset of malignant melanomas, and in a newly delineated deletion syndrome. Here we report that the p36.3 region of human chromosome 1 consists of two identical genomic regions, each of which contain a Cdc2L gene linked to a metalloprotease (MMP) gene in a tail-to-tail configuration. This duplicated genomic region is also linked tightly to D1Z2, a genetic marker containing a highly polymorphic VNTR (variable number tandem repeat) consisting of an unusual 40-bp reiterated sequence. Thus, these genes and the polymorphic marker D1Z2 are organized as follows: telomere-D1Z2-5'-MMP22-3'-3'-Cdc2L2-5'-5'-Cdc2L1 -3'- 3'-MMP21-5' centromere. Remarkably, the introns and exons of Cdc2L1 and Cdc2L2, as well as their flanking regions, are essentially identical. A total of 15 amino acid differences, 12 nonconservative and 3 conservative, can be found in the 773-786 amino acids specified by the various products of the Cdc2L genes. Two separate promoter/5' untranslated (UT) regions, CpG1 and CpG2, are identical to a reported previously methylated genomic CpG sequence and are used to express >20 different Cdc2L transcripts from the two genes. The expression of CpG2 transcripts from Cdc2L1 and Cdc2L2 is tissue/cell-line specific. CpG1 transcripts are expressed ubiquitously from both genes, with perhaps some bias towards the expression of CpG1 Cdc2L1 mRNAs in certain hematopoietic cells. PMID- 9750193 TI - Physical and comparative mapping of distal mouse chromosome 16. 5 p5. AB - Distal mouse Chromosome 16 (Chr. 16) includes a region of conserved linkage with human Chromosome 21 (Chr. 21). Mouse models of Down syndrome based on trisomy of distal Chr. 16 have several phenotypes similar to those seen in human patients and have proven useful for correlating dosage imbalance of specific genes with specific developmental anomalies. The degree to which such findings can be related to Down syndrome depends on how well the conserved synteny is maintained. Twenty-four genes have been mapped in both species and there are no discordancies, but the region could carry hundreds of genes. Comparative sequence represents the ultimate comparative map and will aid in identification of genes and their regulatory sequences. A physical map of the distal 4.5 Mb of Chr. 16 has been assembled as an essential step toward a map of sequence-ready templates. The map consists of 51 YACs and 15 BACs and includes 18 transcripts, 9 of which are mapped for the first time in mouse, and 3 of which are, for the first time, described in either species. YAC fragmentation was used to precisely localize the 49 markers on the map. Comparison of this physical map with that of the corresponding region on Chr. 21 shows conservation not only of gene order but of size in the 3 Mb from Cbr1 to Ets2; distal to Ets2, the human map is expanded. PMID- 9750195 TI - A computer program for aligning a cDNA sequence with a genomic DNA sequence. AB - We address the problem of efficiently aligning a transcribed and spliced DNA sequence with a genomic sequence containing that gene, allowing for introns in the genomic sequence and a relatively small number of sequencing errors. A freely available computer program, described herein, solves the problem for a 100-kb genomic sequence in a few seconds on a workstation. PMID- 9750194 TI - A primary male autosomal linkage map of the horse genome. AB - A primary male autosomal linkage map of the domestic horse (Equus caballus) has been developed by segregation analysis of 140 genetic markers within eight half sib families. The family material comprised four Standardbred trotters and four Icelandic horses, with a total of 263 offspring. The marker set included 121 microsatellite markers, eight protein polymorphisms, five RFLPs, three blood group polymorphisms, two PCR-RFLPs, and one single strand conformation polymorphism (SSCP). One hundred markers were arranged into 25 linkage groups, 22 of which could be assigned physically to 18 different chromosomes (ECA1, ECA2, ECA3, ECA4, ECA5, ECA6, ECA7, ECA9, ECA10, ECA11, ECA13, ECA15, ECA16, ECA18, ECA19, ECA21, ECA22, and ECA30). The average distance between linked markers was 12.6 cM and the longest linkage group measured 103 cM. The total map distance contained within linkage groups was 679 cM. If the distances covered outside the ends of linkage groups and by unlinked markers were included, it was estimated that the marker set covered at least 1500 cM, that is, at least 50% of the genome. A comparison of the relationship between genetic and physical distances in anchored linkage groups gave ratios of 0.5-0.8 cM per Mb of DNA. This would suggest that the total male recombinational distance in the horse is 2000 cM; this value is lower than that suggested by chiasma counts. The present map should provide an important framework for future genome mapping in the horse. PMID- 9750196 TI - Automated sequence preprocessing in a large-scale sequencing environment. AB - A software system for transforming fragments from four-color fluorescence-based gel electrophoresis experiments into assembled sequence is described. It has been developed for large-scale processing of all trace data, including shotgun and finishing reads, regardless of clone origin. Design considerations are discussed in detail, as are programming implementation and graphic tools. The importance of input validation, record tracking, and use of base quality values is emphasized. Several quality analysis metrics are proposed and applied to sample results from recently sequenced clones. Such quantities prove to be a valuable aid in evaluating modifications of sequencing protocol. The system is in full production use at both the Genome Sequencing Center and the Sanger Centre, for which combined weekly production is approximately 100, 000 sequencing reads per week. PMID- 9750198 TI - [Data processing system using PC9800 and Autonics APC204 digitizer]. PMID- 9750199 TI - [Analog-digital conversion using pCLAMP and DOS/V]. PMID- 9750197 TI - Microsatellite marker content mapping of 12 candidate genes for obesity: assembly of seven obesity screening panels for automated genotyping. AB - Twin studies, adoption studies, and studies of familial aggregation indicate that obesity has a genetic component. Whereas, the genetic factors predisposing to obesity have been elucidated for several rare syndromes, the factors responsible for obesity in the general population have remained elusive. Genetic studies of complex traits are often accelerated by the use of candidate genes. To facilitate genetic studies of human obesity, seven multiplex panels of candidate genes for obesity that are suitable for fluorescent genotyping have been assembled. The multiplex panels are composed of 66 microsatellite markers linked tightly to 16 human gene products that are of potential importance in the control of body weight or linked to syndromic forms of obesity. As part of these efforts 12 previously cloned genes have been placed on the human physical map. In addition the chromosomal location of three of these genes, ART, NYP Y6R, and PPARgamma, are reported for the first time. These resources will be of use in studies to identify the genetic factors responsible for human obesity. [Figures are available at http://www.genome.org] PMID- 9750200 TI - The emerging role of estrogen/androgen therapy in the care of the postmenopausal patient. A symposium. PMID- 9750201 TI - Plasma androgens in women. PMID- 9750202 TI - Broadened spectrum of menopausal symptom relief. PMID- 9750203 TI - Effects of estrogens, progestins, and androgens on coronary vasomotion and atherosclerosis. PMID- 9750204 TI - Efficacy and safety of estrogen/androgen therapy. Menopausal symptoms, bone, and cardiovascular parameters. PMID- 9750205 TI - Practical clinical considerations in the use of estrogen/androgen therapy. A case based forum. PMID- 9750207 TI - Transcriptional regulation of photosynthesis operons in Rhodobacter sphaeroides 2.4.1. PMID- 9750206 TI - Isolation and functional study of photosystem I subunits in the cyanobacterium Synechocystis sp. PCC 6803. PMID- 9750208 TI - Copper-responsive gene expression during adaptation to copper deficiency. PMID- 9750209 TI - Gene modifications and mutation mapping to study the function of photosystem II. PMID- 9750210 TI - Department of Biological Sciences, Louisiana State University, Baton Rouge 70803, USA. PMID- 9750211 TI - Deciphering rules of helix stability in peptides. PMID- 9750212 TI - Modified amino acids as probes of helix stability. PMID- 9750213 TI - Measuring hydration changes of proteins in solution: applications of osmotic stress and structure-based calculations. PMID- 9750214 TI - Protein folding in membranes: determining energetics of peptide-bilayer interactions. PMID- 9750215 TI - Tight ligand binding affinities determined from thermodynamic linkage to temperature by titration calorimetry. AB - A general isothermal titration calorimetry method is described that can be used to determine equilibrium binding constants for high-affinity interactions of ligands with biological macromolecules. The method exploits the thermodynamic linkage between the ligand binding equilibrium constant and temperature. By measuring the binding enthalpy change for an interaction as a function of temperature directly, the change in affinity can be calculated with an integrated form of the van't Hoff equation that is applicable to ligand binding to biological macromolecules. When the temperature dependence of the affinity is combined with the absolute affinity determined independently at a convenient temperature (where the affinity can most accurately or most easily be measured), the absolute binding affinity over the entire temperature range is determined. PMID- 9750216 TI - Structure-based prediction of binding affinities and molecular design of peptide ligands. PMID- 9750217 TI - Molecular crowding: analysis of effects of high concentrations of inert cosolutes on biochemical equilibria and rates in terms of volume exclusion. PMID- 9750218 TI - Application of automated methods for determination of protein conformational stability. AB - Automation of protein stability determination saves much time and often results in better data. In our laboratory we estimate that the lack of such instrumentation would increase the amount of time required to make and characterize a mutant protein by at least one-third. As with any experiment, lack of attention to important details can required repeating the experiment or, worse, give superficially good data that are flawed in some manner. Nevertheless, with a minimal investment of attention, an automated instrument can give dramatic gains in productivity. PMID- 9750219 TI - Calculations of proton-binding thermodynamics in proteins. AB - Computational models of proton binding can range from the chemically complex and statistically simple (as in the quantum calculations) to the chemically simple and statistically complex. Much progress has been made in the multiple-site titration problem. Calculations have improved with the inclusion of more flexibility in regard to both the geometry of the proton binding and the larger scale protein motions associated with titration. This article concentrated on the principles of current calculations, but did not attempt to survey their quantitative performance. This is (1) because such comparisons are given in the cited papers and (2) because continued developments in understanding conformational flexibility and interaction energies will be needed to develop robust methods with strong predictive power. Nevertheless, the advances achieved over the past few years should not be underestimated: serious calculations of protonation behavior and its coupling to conformational change can now be confidently pursued against a backdrop of increasing understanding of the strengths and limitations of such models. It is hoped that such theoretical advances will also spur renewed experimental interest in measuring both overall titration curves and individual pKa values or pKa shifts. Exploration of the shapes of individual titration curves (as measured by Hill coefficients and other parameters) would also be useful in assessing the accuracy of computations and in drawing connections to functional behavior. PMID- 9750220 TI - Analysis of spectra from multiwavelength oxygen-binding studies of mixed metal hybrid hemoglobins. PMID- 9750221 TI - Study of the Bohr effect in hemoglobin intermediates. PMID- 9750222 TI - Application of Planck-Benzinger relationships to biology. PMID- 9750223 TI - Kinetic analysis of macromolecular interactions using surface plasmon resonance biosensors. PMID- 9750224 TI - Prediction of binding energetics from structure using empirical parameterization. AB - We have presented an empirical method that can be used to predict the binding energetics for protein-protein or protein-peptide interactions from three dimensional structures. The approach differs from other empirical methods in yielding a thermodynamic description of the binding process, including delta Cp, delta H degree, and delta S degree, rather than predicting delta G degree alone. These thermodynamic terms can provide a wealth of detail about the nature of the interaction, and, if sufficient experimental data are available for comparison, a greater assessment of the accuracy of the calculations. A recurring theme throughout this article is the need for more complete thermodynamic and structural characterizations of protein-ligand interactions. This includes not only characterization of the binding delta H degree, delta S degree, and delta Cp, but a thorough investigation into equilibria linked to binding, such as protonation, ion binding, and conformational changes. Sufficient data will allow parameterization on binding data rather than protein unfolding data. Further inclusion of information obtained from unfolding studies is not likely to generate significant improvement in the accuracy of the calculations. As additional binding data become available, the parameterization can be further extended to include relationships derived from analyses of these data. Not only will this increase accuracy and thus confidence, but allow extension of the method of additional types of interactions. PMID- 9750225 TI - Photoacoustic calorimetry of proteins. AB - We have described two examples of time-resolved photoacoustic calorimetry for the study of heme protein transient intermediates. Before photoacoustic calorimetry, determining thermodynamic information on short-lived intermediates was difficult. Along with being sensitive to enthalpic and volume changes, photoacoustic calorimetry can detect conformational changes in a time-resolved manner. In complex protein systems, the interpretation of the structural origins of a conformational change is sometimes difficult. Site-directed mutagenesis has been used successfully to identify the residues that play important roles in the ligand binding to both Mb and cytochrome P450cam. In both systems the hydration state of salt bridges gave rise to volume changes that were identified through mutagenesis of the residues involved. With its increasing popularity and the power of site-directed mutagenesis, time-resolved photoacoustic calorimetry is fast becoming a technique to probe conformational dynamics in proteins. PMID- 9750226 TI - Isoergonic cooperativity: a novel form of allostery. PMID- 9750227 TI - Theoretical aspects of isothermal titration calorimetry. PMID- 9750228 TI - Use of Poisson-Boltzmann equation to analyze ion binding to DNA. PMID- 9750229 TI - Following the folding of RNA with time-resolved synchrotron X-ray footprinting. AB - The rapid mixing synchrotron X-ray footprinting technique described in this article allows nucleic acid folding and ligand binding reactions to be followed on a millisecond time resolution with single nucleotide resolution. In principle, the change in .OH protection of every nucleotide in a nucleic acid hundreds of nucleotides long can be monitored separately. In addition, a wide range of solution conditions are compatible with the radiolytic generation of .OH. These characteristics of synchrotron X-ray footprinting create opportunities for conducting thermodynamic and kinetic studies of nucleic acids that are both comprehensive and detailed. Kinetic footprinting studies of a number of systems have been initiated by the Center for Synchrotron Biosciences using this technique. PMID- 9750230 TI - Analysis of interactions between CytR and CRP at CytR-regulated promoters. PMID- 9750231 TI - Energetic methods to study bifunctional biotin operon repressor. AB - Application of a broad range of approaches and techniques to analysis of the functional energetics of the biotin regulatory system has enabled dissection of each of the steps in the assembly of this transcriptional repression complex. Although the molecular details of the interactions are not yet completely understood, the studies described in this article have laid a solid foundation for future studies of the system. The application of kinetic and equilibrium methods to studies of binding of the allosteric effector has allow determination of the kinetic parameters governing the interaction of the protein and ligand. The kinetic parameters have, furthermore, been utilized to calculate the equilibrium parameters associated with the binding. The great advantage of using kinetic methods to study the binding process is the additional information provide about the mechanism of allosteric activation of the protein. Based on the initial observation of a kinetic time course that is consistent with the occurrence of a structural change concomitant with effector binding, additional measurements have been performed that have allowed formulation of a testable hypothesis concerning the nature and location of one locus: the structural change in the three-dimensional structure of BirA. Studies of assembly of the protein indicate the bio-5-AMP is an allosteric activator of dimerization of the protein. The dimerization is, however, weak. These results have been critical in analyzing site-specific DNA binding measurements. Application of the DNase I footprinting technique has allowed formulation of a model for association of holoBirA with bioO. Results of studies of binding of the protein to mutant operator templates, although not yielding the anticipated results, provide further insight into the mechanism of association of the protein and DNA. Two models for binding, the validity of which can be tested via the application of kinetic techniques, have been derived from these measurements. The results of quantitative studies of the biotin regulatory system can be interpreted in the context of the biological function of the system. The biotin holoenzyme ligases are a class of enzymes found across the evolutionary spectrum. Only a subset of these enzymes, including BirA, also function as transcriptional repressors. The tight binding of the allosteric effector may be understood in light of the bifunctional nature of the BirA-bio-5'-AMP complex. It is possible that the unusually high thermodynamic and kinetic stability of the complex ensures that the most probable state of the protein in vivo is the adenylate-bound form. This complex, not the unliganded protein, is active in both enzymatic transfer of biotin and site-specific DNA binding. This ensures that on depletion of the intracellular pool of apoBCCP, BirA-bio-5'-AMP accumulates and binds to bioO to repress transcription of the biotin biosynthesis operon. The intracellular demand for and synthesis of biotin are, consequently, tightly coupled in the system. The dimerization that accompanies adenylate binding to BirA appears to be significant for site-specific binding of the protein to bioO. Functionally, the simultaneous binding of the two monomers to the two operator half-sites, regardless of the kinetic mechanism by which it occurs, ensures coordinate regulation of transcription initiation from both biotin operon promoters. The multifaceted approach utilized in studies of the biotin regulatory system can serve as a model for studies of any complex transcriptional regulatory system. It is critical in elucidating the functional energetics of any of these systems that the assembly first be dissected into the constituent interactions and that each of these interactions be studied in isolation. This is not only critical for understanding the physicochemical properties of each individual contributing interaction, but is also a necessary precursor to studies of thermodynamic linkage in the system. (AB PMID- 9750232 TI - Thermal melting properties of C-terminal domain mutants of bacteriophage lambda cI repressor. PMID- 9750233 TI - Investigation of phase transitions in bilayer membranes. AB - This article described three techniques used to study phase transitions in phospholipid bilayers. The complementarity of the three techniques in characterizing the thermotropic and structural properties of phospholipid bilayers has been demonstrated by describing their use to characterize a series of mixed-chain-length PCs. It has been shown that an understanding of the energetics that govern the packing of phospholipid chains in the gel phase can be used to construct a model to interpret thermodynamic data of the PCs. This model, in turn, provided a framework for designing and interpreting the Raman spectroscopic and X-ray diffraction experiments on this series of phospholipids. The result was a complete description of the phase transitions and gel phase packing properties of the mixed-chain-length PCs. The phase diagram of Fig. 5B has been expanded to include the mixed-chain-length PC series C18C18PC through C18C0PC. Furthermore, the phase diagram and the chain inequivalence parameter have been shown to describe the behavior of any mixed-chain-length PC, irrespective of the lengths of the hydrocarbon chains or the position of the chains on the glycerol backbone. This is demonstrated by the additional mixed chain-length PCs plotted in Fig. 5B. With minor modifications, the phase diagram also accurately describes the behavior of mixed-chain-length phosphatidylethanolamines, sphingomyelins, and unsaturated PCs. Finally, it has been demonstrated that a correlation exists between the thermodynamic and the Raman spectroscopic parameters determined for the phase transition of phospholipid bilayers. This correlation is based on the common chain energetics being measured by these two techniques. PMID- 9750234 TI - Membrane-confined analytical electrophoresis. PMID- 9750235 TI - [The nature of the phosphorylation of retinoblastoma gene product in stable mouse and human cell lines]. AB - The retinoblastoma gene product mediates the interaction between transcriptional factors and cyclin-kinase complexes, which perform a regulatory and effector function in the process of cell division. The activity of the retinoblastoma gene product is regulated by phosphorylation, which results in the appearance of additional protein molecules migrating in the region of 105--116 kDa during electrophoresis. Stereochemical analysis has established a direct correspondence between the extent of phosphorylation of the retinoblastoma gene product and its electrophoretic mobility. The results obtained permit an estimate of the phosphorylation of this protein in cells of different tissues by immunoblotting analysis of their lysates. The results of this study demonstrate that the degree of phosphorylation of retinoblastoma gene product in a series of stable cell lines increases as we go from monolayer to multilayer cultures, and further to cell cultures in suspension. Human cells produce more phosphorylated proteins compared to homologous mouse cells. The phosphorylation pattern of retinoblastoma gene product is probably tissue-specific. Much like mouse fibroblasts, HeLa cells may contain a hypophosphorylated protein with a molecular weight of 105 kDa. Phosphorylation of the retinoblastoma gene product in cells of embryonic mouse adenocarcinoma (line p19) changes in response to cloning or stimulation of differentiation by retinoic acid. The formation of all forms of retinoblastoma gene product, which pre-exist during asynchronous growth, is increased in the course of cloning. Differentiation is associated with the synthesis of an increased amount of hypophosphorylated protein. The results obtained lead to the hypothesis that even though the phosphorylation pattern of the retinoblastoma gene product is tissue-specific, it can show significant variation under the conditions of either cloning or differentiation and can be maintained for extended period of time at a new level that was not characteristic of the initial cell population. PMID- 9750236 TI - [The biopsy of preimplantation embryos obtained from donor pigs with follicle stimulating hormone-induced superovulation]. AB - We have examined possible use of the follicle-stimulating hormone (FSH) for the induction of superovulation in pigs and studied the effect of biopsy of preimplantation pig embryos on their survival in vitro. Superovulation was induced by injecting FSH twice daily over a period of four days for a total dose of 25 units. per animal. Pigs receiving pregnant mare serum gonadotropin (PMSG) according to the standard scheme served as the control. The experiments demonstrated that FSH produces significantly better estrus figures as compared with PMSG (100% and 60%, respectively; p < 0.05). The mean number of ovulations per donor was also better in the FSH group, i.e., 36.5 as compared with 17.3 in the control group (p < 0.05). The mean number of embryos obtained from one donor was higher in FSH treated animals as compared with the animals that received PMSG (29.2 vs. 19.3), however, this difference was not statistically significant. Biopsy of preimplantation pig embryos was conducted by a "manual" technique without a micromanipulator; middle blastocysts collected at days 5-6 after insemination were the most convenient for this operation. Embryos at these developmental stages contained a well developed inner cell mass, which allows separation of trophoectodermal cells only, thus minimizing damage to the embryo. The number of cells in dissected fragments did not depend on the stage of development, and even smallest fragments (4 blastomeres) were sufficient for genome analysis with the aid of PCR. The survival of embryos in vitro after biopsy practically did not differ from that of control intact embryos. Thus, we demonstrated the effective use of FSH for the induction of superovulation in pigs; we also determined the developmental stages which are most convenient for conducting biopsies and developed a technique for preimplantation biopsy, which allows genome analysis of embryos without any decrease of their survival in vitro. PMID- 9750237 TI - [Vladimir Vladimirovich Sakharov--an outstanding disciple of N. K. Kol'tsov]. AB - Vladimir Vladimirovich Sakharov would have celebrated his 95th birthday on the 28th of February, 1997. The scientific community celebrated this anniversary by conducting the XXVI Sakharov Lecture. Sacharov Lectures were established in 1969 by the order of the Presidium of the Moscow Society of Naturalists. The XXVI Sakharov Lecture was read at Kol'tsov Institute of Developmental Biology. The session was chaired by Professor N. G. Khrushchov, director of the institute, who has highlighted the achievements of Dr. Sakharov. Professor Komissarov from the Institute of Chemical Physics gave a lecture entitled "A Novel Concept of Photosynthesis and Prebiological Evolution."" PMID- 9750238 TI - Statistical methods for assessing differential vaccine protection against human immunodeficiency virus types. AB - The human immunodeficiency virus type 1 (HIV-1) is extremely diverse. In assessing the utility of an HIV-1 vaccine, an important issue is the possibility of differential protection. We discuss statistical methods of inferring how the vaccine efficacy may vary with viral type from data that would be collected from a randomized, double-blind, placebo-controlled preventive vaccine efficacy trial. Detailed characterization of virus isolated from individuals infected during the trial will be available. We focus on the highly simplified case in which the viral characteristics are summarized by a single feature, which may be nominal, or a scalar quantity that represents distance between the isolate and the prototype virus or viruses used in the vaccine preparation. We consider discrete categorical and continuous response models for this quantity and identify models whose parameters can be interpreted as log ratios of strain-specific relative risks of infection in a prospective model for HIV-1 exposure and transmission. Methods of inference are described for the multinomial logistic regression (MLR) model for discrete categorical response, and a new semiparametric model which can be viewed as a continuous analog of the MLR model is introduced. The methods are illustrated by application to HIV-1 and hepatitis B vaccine trial data. PMID- 9750239 TI - Estimating a distribution function of the tumor size at metastasis. AB - In studying the relationship between the size of primary cancers and the occurrence of metastases, two quantities are of prime importance. The first is the distribution of tumor size at the point of metastatic transition, while the second is the probability that detectable metastases are present when cancer comes to medical attention. Kimmel and Flehinger (1991, Biometrics 47, 987-1004) developed a general nonparametric model and studied its two limiting cases. Because of unidentifiablity of their general model, a new identifiable model is introduced by making the hazard function for detecting a metastatic cancer a constant. The new model includes Kimmel and Flehinger's (1991) second limiting model as a special case. An estimator of the tumor size distribution at metastases is proposed. The result is applied to a set of colorectal cancer data. PMID- 9750242 TI - A semiparametric Bayesian approach to the random effects model. AB - In longitudinal random effects models, the random effects are typically assumed to have a normal distribution in both Bayesian and classical models. We provide a Bayesian model that allows the random effects to have a nonparametric prior distribution. We propose a Dirichlet process prior for the distribution of the random effects; computation is made possible by the Gibbs sampler. An example using marker data from an AIDS study is given to illustrate the methodology. PMID- 9750243 TI - A more flexible regression-to-the-mean model with possible stratification. AB - We consider a regression-to-the-mean model that includes both additive and multiplicative treatment effects. We allow either or both of these treatment effects to be stratified by ranges of the first measurement. We focus on the situation where there is a very large sample on the first measurement and a relatively small subsample for the second measurement is selected, which often occurs in screening trials. We propose some asymptotically efficient estimators for the parameters of the model that are very simple to compute. We begin with a discussion of the full model, and more on tests and estimation for reduced models follows. An example from a large screening trial is discussed. PMID- 9750244 TI - Assessing the sensitivity of regression results to unmeasured confounders in observational studies. AB - This paper presents a general approach for assessing the sensitivity of the point and interval estimates of the primary exposure effect in an observational study to the residual confounding effects of unmeasured variable after adjusting for measured covariates. The proposed method assumes that the true exposure effect can be represented in a regression model that includes the exposure indicator as well as the measured and unmeasured confounders. One can use the corresponding reduced model that omits the unmeasured confounder to make statistical inferences about the true exposure effect by specifying the distributions of the unmeasured confounder in the exposed and unexposed groups along with the effects of the unmeasured confounder on the outcome variable. Under certain conditions, there exists a simple algebraic relationship between the true exposure effect in the full model and the apparent exposure effect in the reduced model. One can then estimate the true exposure effect by making a simple adjustment to the point and interval estimates of the apparent exposure effect obtained from standard software or published reports. The proposed method handles both binary response and censored survival time data, accommodates any study design, and allows the unmeasured confounder to be discrete or normally distributed. We describe applications on two major medical studies. PMID- 9750245 TI - Approaches for optimal sequential decision analysis in clinical trials. AB - Unlike traditional approaches, Bayesian methods enable formal combination of expert opinion and objective information into interim and final analyses of clinical trial data. However, most previous Bayesian approaches have based the stopping decision on the posterior probability content of one or more regions of the parameter space, thus implicitly determining a loss and decision structure. In this paper, we offer a fully Bayesian approach to this problem, specifying not only the likelihood and prior distributions but appropriate loss functions as well. At each data monitoring point, we enumerate the available decisions and investigate the use of backward induction, implemented via Monte Carlo methods, to choose the optimal course of action. We then present a forward sampling algorithm that substantially eases the analytic and computational burdens associated with backward induction, offering the possibility of fully Bayesian optimal sequential monitoring for previously untenable numbers of interim looks. We show that forward sampling can always identify the optimal sequential strategy in the case of a one-parameter exponential family with a conjugate prior and monotone loss functions as well as the best member of a certain class of strategies when backward induction is infeasible. Finally, we illustrate and compare the forward and backward approaches using data from a recent AIDS clinical trial. PMID- 9750246 TI - Estimating a treatment effect from multidimensional longitudinal data. AB - Multidimensional longitudinal data result when researchers measure an outcome through time that is quantified by many different response variables. These response variables are often defined on different numerical scales. The objective of this paper is to present a method to summarize and estimate an overall treatment effect from this type of longitudinal data. A regression model is proposed that assumes the treatment effect can be parameterized as an acceleration or deceleration of the time scale of each response variable's trajectory. Generalized estimating equations are used to estimate the model parameters. Cognitive and functional ability data from Alzheimer's disease patients and quality of life data from an AIDS clinical trial are used to illustrate the model. PMID- 9750247 TI - Age-dependent U-shaped risk functions and Aalen's additive risk model. AB - Epidemiologic studies of time to some failure often show a quadratic relation between the risk of failure and covariate(s). We study the nadir for a given covariate, i.e., the value of the covariate associated with the lowest risk (supposing a U-shape), within Aalen's additive risk model. This model was applied since the effect of the covariate(s) is allowed to vary over time and, as a consequence, a given nadir can vary over time. We propose a test for the null hypothesis that the nadir is time independent and, if this is the case, an estimate of the nadir. Large sample properties of the test statistic and estimator are derived. The methods are illustrated with data where time to death is related to body mass index. PMID- 9750248 TI - Estimating equations with incomplete categorical covariates in the Cox model. AB - Incomplete covariate data is a common occurrence in many studies in which the outcome is survival time. When a full likelihood is specified, a useful technique for obtaining parameter estimates is the EM algorithm. We propose a set of estimating equations to estimate the parameters of Cox's proportional hazards model when some covariate values are missing. These estimating equations can be solved by an algorithm similar to the EM algorithm. Because of the computational burden of finding a solution to these estimating equations, we propose obtaining parameter estimates via Monte Carlo methods. Asymptotic variances of the parameter estimates are also derived. We present a clinical trials example with three covariates, two of which have some missing values. PMID- 9750249 TI - Analysis of age of onset data from case-control family studies. AB - Age of onset data from case-control family studies are frequently used to study the relationship between disease and environmental or genetic factors. Dependence of age of onset within family and case-control sampling designs must be taken into account in analyzing such data. In this paper, we propose a parametric likelihood approach to study the relationship between disease risk and covariates, allowing for any correlations in age of onset of disease between family members and to study the familial association in age of onset of disease due to genetic factors, after adjusting for other shared covariates within the family. The method provides a way to combine the information relating disease incidence to risk factors in relatives with the information contained in the case/control contrasts in order to obtain more precise estimates of the effects of the putative risk factors. A data set from a case-control family study of lung cancer is used to illustrate the method. The analysis indicated that history of smoking, passive smoking, and history of chronic obstructive pulmonary disease are positively associated with lung cancer risk, and that, after adjustment for these risk factors, there is little evidence of familial aggregation in lung cancer risk. PMID- 9750250 TI - Some scale estimators and lack-of-fit tests for the censored two-sample accelerated life model. AB - Some new scale estimators for the censored two-sample accelerated life model are introduced. They are zeros of some integrated weighted difference between the two cumulative hazard estimators. These estimators are asymptotically normal. The weight is chosen to result in estimators whose asymptotic variances do not involve the destiny functions and can be easily estimated. This provides a fast and simple means of statistical inference in the censored two-sample accelerated life model. Through investigating the asymptotic relative efficiency at some important censoring submodels and the finite example behaviors in various numerical studies, we obtain some estimators with very competitive performance. From the new class of scale estimators, some lack-of-fit tests for the accelerated life model are also derived. These tests are related to Gill Schumacher type tests and require little extra computing time once the estimator is obtained. The estimators and tests are illustrated in two applications. For a vaginal cancer data set for rats, the effect of pretreatment regime was found to be well described by the two-sample accelerated life model. For a data set on progression of ovarian cancer, it was found that the effect of grade of disease could not be described either by the two-sample proportional hazards model or the two-sample accelerated life model. PMID- 9750251 TI - Estimating survival curves with left-truncated and interval-censored data under monotone hazards. AB - We show that the nonparametric maximum likelihood estimator (NPMLE) of a survival function may severely underestimate the survival probabilities at very early times for left-truncated and interval-censored data. As an alternative, we propose to compute the (nonparametric) MLE under a nondecreasing hazard assumption, the monotone MLE, by a gradient projection algorithm when the assumption holds. The projection step is accomplished via an isotonic regression algorithm, the pool-adjacent-violators algorithm. This gradient projection algorithm is computationally efficient and converges globally. Monte Carlo simulations show superior performance of the monotone MLE over that of the NPMLE in terms of either bias or variance, even for large samples. The methodology is illustrated with the application to the Wisconsin Epidemiological Study of Diabetic Retinopathy data to estimate the probability of incidence of retinopathy. PMID- 9750252 TI - Construction of a continuous stopping boundary from an alpha spending function. AB - Lan and DeMets (1983, Biometrika 70, 659-663) proposed a flexible method for monitoring accumulating data that does not require the number and times of analyses to be specified in advance yet maintains an overall Type I error, alpha. Their method amounts to discretizing a preselected continuous boundary by clumping the density of the boundary crossing time at discrete analysis times and calculating the resultant discrete-time boundary values. In this framework, the cumulative distribution function of the continuous-time stopping rule is used as an alpha spending function. A key assumption that underlies this method is that future analysis times are not chosen on the basis of the current value of the statistic. However, clinical trials may be monitored more frequently when they are close to crossing the boundary. In this situation, the corresponding continuous-time boundary should be used. Here we demonstrate how to construct a continuous stopping boundary from an alpha spending function. This capability is useful in the design of clinical trials. We use the Beta-Blocker Heart Attack Trial (BHAT) and AIDS Clinical Trials Group protocol 021 for illustration. PMID- 9750253 TI - A method for sequential analysis of survival data with nonproportional hazards. AB - Two tests are proposed for comparing the survival curves of patients randomised between an experimental treatment and a control treatment when it is anticipated that the two survival curves may not satisfy the assumption of proportional hazards. The tests are particularly useful for the situation in which the survival curves are coincident or cross over early in the follow-up period and then diverge. The tests compare the probabilities of survival for longer than some fixed time since randomisation for the two groups of patients. Both methods take account of the right-censored observations, and both are associated with methods for estimating and setting confidence limits for treatment differences. The first method is a mathematically direct approach based on the derivation of the efficient score statistic and Fisher's information. The second method is simpler, being based on Kaplan-Meier estimates and their variances. Conventional methods of sample size determination require the assumption of proportional hazards. Here a sequential approach is used, as it is difficult to set the sample size in advance without strong assumptions about the relationship between the two survival curves. Simulation results giving information on the size and power of the proposed tests are provided and the tests are applied to data from a clinical trial in breast cancer. PMID- 9750255 TI - Minimum distance estimation of mutational parameters for quantitative traits. AB - Individual spontaneous mutations affecting the expression of quantitative traits cannot be systematically identified and, therefore, their effect on the trait cannot be measured. Thus, the rate of occurrence of such mutations and the moments of the probability distribution of the corresponding effects, which are important in evolutionary studies, remain unknown. Here we propose a method to estimate those mutational properties from the observed distribution of the trait mean in a set of independent inbred lines (all derived from the same homozygous base population) in which mutations had been allowed to accumulate randomly. It is based on the use of the well-known minimum distance method, i.e., on the minimization of a distance between the observed distribution and that expected on the basis of a genetic model. We analyze data for three morphological traits (wing length and abdominal and sternopleural bristle number) in Drosophila melanogaster. The method appears to be powerful, giving evolutionary coherent estimates of relevant mutational properties that had not been estimated previously. For all traits, mutational rates were low (smaller than 0.05). Most mutations affecting wing length or abdominal bristle number and negative effect, while almost half of those affecting sternopleural bristle number had positive effect. For each trait, results obtained from data on different generations are in qualitative agreement, although mutational effects seem to depend on generation-specific environmental factors. The method detected between-trait differences in the kurtosis coefficient of the distribution of mutational effects, which varied from values close to that of the normal distribution (wing length) to relatively high values (sternopleural bristle number). It reveals that an important proportion of the mutational input variance of each trait is due to mutations with absolute effect smaller than 0.5 environmental standard deviation units. For morphological traits undergoing weak direct selection, this suggests that large amounts of genetic variance due to genes segregating at intermediate frequencies can be present at the equilibrium. PMID- 9750256 TI - Modeling multivariate discrete failure time data. AB - A bivariate discrete survival distribution that allows flexible modeling of the marginal distributions and yields a constant odds ratio at any grid point is proposed. The distribution can be extended to a multivariate distribution and is readily generalized to accommodate covariates in the marginal distributions and pairwise odds ratios. In addition, a pseudo-likelihood estimation procedure for estimating the regression coefficients in the marginal models and the association parameters in the pairwise odds ratios is presented. We evaluate the performance of the proposed estimation procedure through simulations. For bivariate data, pseudo-likelihood estimation of the association parameter has high efficiency. Loss of efficiency in the marginal regression coefficient estimates is small when the association is not strong. For both the marginal regression coefficients and the association parameter, coverage probabilities are close to the 95% nominal level. For multivariate data, the simulation results show that the parameter estimates are consistent. Coverage probability for the regression coefficient in the marginal model is close to the 95% nominal level but is slightly less than the nominal level for the association parameter. We illustrate the proposed methods using a subset of the Framingham Heart Study data where a significant positive association was found between the failure times of siblings. PMID- 9750257 TI - An order-directed score test for trend in ordered 2 X K tables. AB - This paper presents a new test procedure for detecting trend in ordered 2 X K tables. Using an order-directed score statistic, the procedure does not require a set of scores preassigned to the ordinal categories under consideration. Thus the problem of varying p-values of linear rank tests, due to choices of different scoring systems, is avoided. The proposed test procedure can be easily generalized to handle stratified analysis where data are represented by several 2 x K tables. Examples are given to illustrate the method. PMID- 9750258 TI - Optimal two-stage design for a series of pilot trials of new agents. AB - An approach to determine the appropriate sample sizes for a series of screening trials to identify promising new therapeutic agents was presented by Yao, Begg, and Livingston (1996, Biometrics 52, 992-1001). This approach is now improved to a two-stage design that further minimizes the time to identify a promising agent under fixed error rates. When applied to data from the historical experience of exploratory vaccination trials at Memorial Sloan-Kettering Cancer Center, the method demonstrates that relatively small individual screening trials are optimal. The reliability of the results is evaluated using the bootstrap. PMID- 9750259 TI - Cancer mortality around French nuclear sites. PMID- 9750260 TI - New perspectives on mate choice and the MHC. AB - A long series of studies on mice has shown that mate choice decisions can be made on the basis of individual genotype at the major histocompatibility complex (MHC), which accords well with the importance of immunocompetence in some theories of sexual selection. Recent work on other vertebrate species, including humans, indicates that MHC-based mate choice is not restricted to the genus Mus. However, its importance may vary among species as a result of differences in social and mating system structure, and perhaps genome structure. There appears to be a general preference expressed for MHC-dissimilar mates, and such MHC disassortative mating may be involved in maintaining MHC and/or genome-wide diversity in natural populations. The strength and direction of MHC-based mating preference can vary, and may be modulated by factors such as genetic background, sex, and early life experience. PMID- 9750261 TI - Reaction norm functions and QTL-environment interactions for flowering time in Arabidopsis thaliana. AB - Many plant traits are phenotypically plastic in response to resource levels that vary continuously among environments. To be able to predict phenotypes in new environments, it is useful to model reaction norms as functions, rather than as a collection of discrete character states. Flowering date and rosette leaf number were measured in 100 recombinant inbred lines of Arabidopsis thaliana, grown on a gradient of light intensity. The results show that there is genetic variation among the recombinant inbred lines for parameters of the reaction norm functions. Genetic variances for leaf number and flowering date are highest under low light conditions. Underlying quantitative trait loci (QTLs) affecting the shape of the reaction norm functions were mapped by modifying Haley & Knott (1992) regressions to include polynomial effects of the environment. Quantitative trait loci of large effect were generally insensitive to the resource gradient. Seven QTLs affecting flowering date and eight QTLs for rosette leaf number were identified, of which only two had significant effects on the linear and quadratic components of the reaction norm function. These results suggest that the genotype environment interactions for flowering time are controlled by many minor genes, whose effects are below the detection limit in most mapping experiments. PMID- 9750262 TI - Origin and evolution of twin microsatellites in the genus Oryza. AB - An ancestral sequence of twin microsatellites of rice was found in a wild species. Twin microsatellite loci, RM20A and RM20B, were located on separate regions of chromosomes 11 and 12, which had been duplicated during rice evolution. These twin microsatellites showed different allele diversities in A genome species of the genus Oryza. This difference was caused by repetition of a simple sequence consisting of (TAA)n. Oryza longistaminata contains a short poly(A) sequence in this region instead of the poly(TAA) found in other species. A sequence comparison of RM20-related amplicons suggested that the poly(A) containing sequence is the ancestral sequence of the RM20A and RM20B microsatellites. A simple base substitution in the poly(A) sequence may have produced the longer microsatellite motif (TAA). This mutation may have occurred on one of the chromosomes of a hypothetical ancestor of the A genome species before duplication of the chromosome segments. PMID- 9750263 TI - Mapping quantitative trait loci for ordered categorical traits in four-way crosses. AB - Many quantitative traits of economical importance are ordinal in nature. Although methods of mapping quantitative trait loci (QTLs) for continuous quantitative characters are well developed, such methods for ordinal characters are generally lacking. In this paper, we develop a method based on the framework of a generalized linear model using four-way cross populations. The method estimates and tests the average effects of a gene substitution in the parents. All markers in the same linkage group are simultaneously used to infer the allelic transmission of a putative QTL. General results of the method are demonstrated by a few simulation experiments. We discuss extensions of the method to QTL mapping in full-sib families. PMID- 9750264 TI - Autoregulation of angiogenesis by cells of the vessel wall. AB - The cells of the vessel wall can regulate angiogenesis by producing growth factors, proteolytic enzymes, extracellular matrix components, cell adhesion molecules, and vasoactive factors. This property enables preexisting blood vessels to generate new vessels in the absence of exogenous angiogenic stimuli. Vascular autoregulation of angiogenesis can be studied by culturing rat aortic or venous explants in collagen gels under serum-free conditions. In this system, the combined effect of injury and exposure of explants to collagen triggers a self limited angiogenic response. Interactions among endothelial cells, smooth muscle cells, and fibroblasts play a critical role in the regulation of this process. This chapter reviews the literature on angiogenesis, focusing on the vessel wall as a highly specialized and plastic tissue capable of regenerating itself through autocrine, paracrine, and juxtacrine mechanisms. PMID- 9750265 TI - Fibroblast growth factors as multifunctional signaling factors. AB - The fibroblast growth factor (FGF) family consists of at least 15 structurally related polypeptide growth factors. Their expression is controlled at the levels of transcription, mRNA stability, and translation. The bioavailability of FGFs is further modulated by posttranslational processing and regulated protein trafficking. FGFs bind to receptor tyrosine kinases (FGFRs), heparan sulfate proteoglycans (HSPG), and a cysteine-rich FGF receptor (CFR). FGFRs are required for most biological activities of FGFs. HSPGs alter FGF-FGFR interactions and CFR participates in FGF intracellular transport. FGF signaling pathways are intricate and are intertwined with insulin-like growth factor, transforming growth factor beta, bone morphogenetic protein, and vertebrate homologs of Drosophila wingless activated pathways. FGFs are major regulators of embryonic development: They influence the formation of the primary body axis, neural axis, limbs, and other structures. The activities of FGFs depend on their coordination of fundamental cellular functions, such as survival, replication, differentiation, adhesion, and motility, through effects on gene expression and the cytoskeleton. PMID- 9750266 TI - Structural and functional characteristics of the centrosome in gametogenesis and early embryogenesis of animals. AB - We present a description of the wide spectrum of centrosome behavior during gametogenesis, early development, and cell differentiation. During meiosis and terminal differentiation of gametes there occurs a process of centrosome maturation which includes alterations in characteristics such as the number of centriolar cylinders and their structure if the basal body is formed and ability to function as MTOC, reduplicate, split, and serve as a polar organizer. Such centrosome properties require modifications of the molecular composition. Maturation of the centrosome in gametes may be compared to transformation of centrosome characteristics during terminal differentiation of other cells. After fertilization different properties of maternal and paternal centrosomes are supposed to combine, adding to each other in the fused (hybrid) centrosome of a zygote. Restoration of centrosome features typical in diploid somatic cells takes place in cells of a developing embryo in the course of early cell cycles. PMID- 9750267 TI - Cell and molecular biology of the pars tuberalis of the pituitary. AB - The pars tuberalis of the adenohypophysis is mainly composed of a special type of endocrine cells, pars tuberalis-specific cells, lining the primary capillary plexus of the hypophysial portal system. Dense expression of melatonin receptors and marked changes in morphological appearance, production pattern, and secretory activity during annual cycle show that these cells are highly sensitive to changes in photoperiod. This leads to the hypothesis that the pars tuberalis is involved in the transmission of photoperiodic stimuli to endocrine targets. Several investigations support the theory that pars tuberalis-specific cells are multipotential cells exerting a modulatory influence on the secretory activity of the pars distalis. Specifically, there is accumulating evidence that seasonal modulation of prolactin secretion, independent of hypothalamic input, is due to melatonin-regulated activity of pars tuberalis-specific cells. The exact nature of secretory products and their effects within neuroendocrine regulation, however, remain rather enigmatic. Accordingly, molecular mechanisms regulating gene expression under the influence of photoperiod, respectively, circulating melatonin levels are still incomplete. Recent cloning of melatonin receptor genes and new data on intracellular signal transduction will probably lead to new insights on melatonin action and pars tuberalis-specific cell physiology. PMID- 9750268 TI - Polarization of the Na+, K(+)-ATPase in epithelia derived from the neuroepithelium. AB - The neuroepithelium generates a fascinating group of epithelia. One of their intriguing properties is how they polarize the distribution of the Na+, K(+) ATPase. Typically, this ion pump is concentrated in the basolateral membrane, but it is concentrated in the apical membranes of the retinal pigment epithelium and the epithelium of the choroid plexus. A comparison of their development with that of systemic epithelia yields insights into how cells polarize the distribution of this and other membrane proteins. The polarization of the Na+, K(+)-ATPase depends upon the interplay between different sorting signals and different types of polarity mechanisms. These include intracellular targeting signals that direct the delivery of newly synthesized proteins, and maintenance signals that stabilize proteins in the proper membrane domain. Conflicting signals appear to be arranged in a hierarchy that can be rearranged as cells respond to certain environmental stimuli. Part of this response is mediated by changes in the distribution and composition of the cortical cytoskeleton. PMID- 9750269 TI - Desmosomes: intercellular adhesive junctions specialized for attachment of intermediate filaments. AB - Cell-cell adhesion is thought to play important roles in development, in tissue morphogenesis, and in the regulation of cell migration and proliferation. Desmosomes are adhesive intercellular junctions that anchor the intermediate filament network to the plasma membrane. By functioning both as an adhesive complex and as a cell-surface attachment site for intermediate filaments, desmosomes integrate the intermediate filament cytoskeleton between cells and play an important role in maintaining tissue integrity. Recent observations indicate that tissue integrity is severely compromised in autoimmune and genetic diseases in which the function of desmosomal molecules is impaired. In addition, the structure and function of many of the desmosomal molecules have been determined, and a number of the molecular interactions between desmosomal proteins have now been elucidated. Finally, the molecular constituents of desmosomes and other adhesive complexes are now known to function not only in cell adhesion, but also in the transduction of intracellular signals that regulate cell behavior. PMID- 9750270 TI - Genotypic heterogeneity among Lactobacillus helveticus strains isolated from natural cheese starters. AB - A total of 23 strains of Lactobacillus helveticus isolated from natural whey starter cultures for Italian hard cheeses and three reference strains were characterized by plasmid profiling, ribotyping and random amplified polymorphic DNA (RAPD) fingerprinting. The data showed an interesting strain heterogeneity in natural cheese starters, that seemed not only strain-dependent, but also related to the source of isolates. Nineteen of the strains tested harboured extrachromosomal elements, whilst 11 different plasmid profiles were detected. Ribotyping with a variety of restriction enzymes differentiated 11 strains and in a few cases, RAPD fingerprinting allowed differentiation amongst strains that were not distinguished by the other two techniques. PMID- 9750272 TI - Reliability of CHROMagar O157 for the detection of enterohaemorrhagic Escherichia coli (EHEC) O157 but not EHEC belonging to other serogroups. AB - CHROMagar O157 is designed for the rapid isolation and identification of enterohaemorrhagic Escherichia coli (EHEC), particularly O157, characterized by pink colonies. Five hundred and eighty-five E. coli strains, including O157, O111 and O113 serogroups, from many sources were examined on CHROMagar O157. Enterohaemorrhagic E. coli O157 could readily be isolated and recognized uniquely by typical pink colonies. Some other EHEC also produced pink colonies, whereas O113 and many other EHEC strains were blue and indistinguishable from Shiga-like toxin-negative strains of E. coli. PMID- 9750271 TI - A reporter system for analysis of regulatable promoter functions in the basidiomycete fungus Phanerochaete chrysosporium. AB - To establish a system for functional analysis of regulatable gene promoters involved in lignocellulose degradation by Phanerochaete chrysosporium, a DNA construct was made in which a minimal promoter region, 410 bp of the 5' untranslated region (UTR) of the cbhI.1 gene of P. chrysosporium, was fused to the phlR sequence of Streptoallotiechus hindustanus. This construct was used to transform P. chrysosporium to phleomycin resistance. Southern blot analysis revealed that the incoming DNA was maintained extrachromosomally in the transformants. The donor DNA was methylated by the fungus; inhibition of such methylation allowed transformants to be distinguished from 'false-positive' phlR colonies. Reverse transcriptase-polymerase chain reaction analysis of gene expression revealed that the cbhI.1 5' UTR/phlR sequence construct was transcribed, but that an intron within the cbhI.1 promoter was not excised from transcripts of the transforming DNA construct. Moreover, catabolite repression of cbhI.1 expression is relaxed in the construct. These observations suggest that normal function of the cbhI.1 promoter requires more than 410 bp. PMID- 9750274 TI - Decontamination of drinking water by direct heating in solar panels. AB - A device was developed for direct heating of water by solar radiation in a flow through system of copper pipes. An adjustable thermostat valve prevents water below the chosen temperature from being withdrawn. The results show that it is possible to eliminate coliform and thermotolerant coliform bacteria from naturally contaminated river water by heating to temperatures of 65 degrees C or above. Artificial additions of Salmonella typhimurium, Streptococcus faecalis and Escherichia coli to contaminated river water were also inactivated after heating to 65 degrees C and above. The total viable count could be reduced by a factor of 1000. The heat-resistant bacteria isolated from the Mlalakuva River (Tanzania) were spore-forming bacteria which exhibited greater heat resistance than commonly used test bacteria originating from countries with colder climates. To provide a good safety margin it is recommended that an outlet water temperature of 75 degrees C be used. At that temperature the daily production was about 501 of decontaminated water per m2 of solar panel, an amount that could be doubled by using a heat exchanger to recycle the heat. PMID- 9750273 TI - The use of a ribosomal RNA targeted oligonucleotide probe for fluorescent labelling of viable Cryptosporidium parvum oocysts. AB - A fluorescence in situ hybridization (FISH) technique has been developed for the fluorescent labelling of Cryptosporidium parvum oocysts in water samples. The FISH technique employs a fluorescently labelled oligonucleotide probe (Cry1 probe) targeting a specific sequence in the 18S ribosomal RNA (rRNA) of C. parvum. Hybridization with the Cry1 probe resulted in fluorescence of sporozoites within oocysts that were capable of excystation, while oocysts that were dead prior to fixation did not fluoresce. Correlation of the FISH method with viability as measured by in vitro excystation was statistically highly significant, with a calculated correlation coefficient of 0.998. Examination of sequence data for Cryptosporidium spp. other than C. parvum suggests that the Cry1 probe is C. parvum-specific. In addition, 19 isolates of C. parvum were tested, and all fluoresced after hybridization with the Cry1 probe. Conversely, isolates of C. baileyi and C. muris were tested and found not to fluoresce after hybridization with the Cry1 probe. The fluorescence of FISH-stained oocysts was not bright enough to enable detection of oocysts in environmental water concentrates containing autofluorescent algae and mineral particles. However, in combination with immunofluorescence staining, FISH enabled species-specific detection and viability determination of C. parvum oocysts in water samples. PMID- 9750275 TI - Growth characteristics and metabolic activities of the methanotrophic heterotrophic groundwater community. AB - In this work the growth characteristics and metabolic activities of the methanotrophic-heterotrophic groundwater community (culture MM1) as well as of individual community members were studied. When growing in shake flasks, under various methane and oxygen tensions, culture MM1 revealed the capability of a stable association consisting of one obligate methanotroph with type II intracytoplasmic membranes as the dominant strain, and four or five heterotrophs of different morphological, physiological and metabolic characteristics. Coexistence of different populations and the stability of culture MM1 under various conditions suggested that complex relationships may exist between the community members. Most of these relationships seem to be beneficial for both the methanotroph and heterotrophs, making the community adaptable to a range of environmental conditions containing methane as the only carbon source. Furthermore, faster and more complete transformation of 2-[4 (sulphophenyl)]decane (2C10LAS) by the community than by any of the community members alone, illustrates the role and importance of methanotrophic heterotrophic interactions in combined metabolic attack on complex linear alkylbenzenesulphonates molecules. PMID- 9750276 TI - Generation of a reproducible nutrient-depleted biofilm of Escherichia coli and Burkholderia cepacia. AB - An in vitro method of growing bacteria as a defined nutrient-depleted biofilm is proposed. The medium was defined nutritionally in terms of the quantitative composition and by the total amount of nutrient required to achieve a defined population size. Escherichia coli and Burkholderia cepacia were incubated on a filter support placed on a defined volume of solid medium. The change of biomass of the biofilm population was compared with the change in a planktonic culture. The size of the population in stationary phase was proportional to the concentration of limiting substrate up to 40 mumol cm-1 glucose for E. coli and up to 2.7 x 10(-9) mol cm-2 iron for B. cepacia. Escherichia coli growing exponentially had a growth rate of mu = 0.30 h-1 in a biofilm and mu = 0.96 h-1 in planktonic culture. The growth rate, mu, for exponentially growing B. cepacia in a biofilm was 1.12 h-1 and in planktonic culture 0.78 h-1. This method allows the limitation of the size of a biofilm population to a chosen value. PMID- 9750277 TI - Quantitative detection of Sphingomonas chlorophenolica in soil via competitive polymerase chain reaction. AB - The 16S ribosomal RNA gene sequence of the pentachlorophenol degrader Sphingomonas chlorophenolica strain RA2 was used to generate specific polymerase chain reaction (PCR) primers for the detection of this strain in soil, whereas a region internal to the two primers was used to provide an S. chlorophenolica strain RA2-specific oligonucleotide probe. The PCR detection system resulted in a 727 bp product detectable via gel electrophoresis and hybridization. It was specific for strain RA2 and its close relative, S. chlorophenolica ATCC 39723, as evidenced by PCR amplifications of a range of bacterial genomic DNAs. Tests of total microbial community DNA obtained from five uninoculated and two RA2 inoculated soils confirmed this specificity for introduced S. chlorophenolica RA2. Strain RA2 could be detected in soil down to a level of 10(3) cfu g-1 soil. Two strategies were followed to generate internal standard DNA for competitive PCR. First, a 479 bp MIMICS fragment was obtained based on a previously constructed gene cassette; however, this standard did not reliably quantify RA2 targets. Low stringency PCR performed with a range of bacterial genomic DNAs resulted in the generation of an amplicon with a Paenibacillus azotofixans strain that was slightly smaller than the RA2-derived product. Both products were easily separable via conventional gel electrophoresis. The use of this competitor in a threefold dilution scheme applied to the target DNA allowed for the quantitative detection of RA2-specific target DNA molecules from pure culture and from soil. The fate of strain RA2 in pentachlorophenol-contaminated soil was described using this competitive PCR approach, and the organism was shown to persist at two inoculum levels over prolonged periods of time. PMID- 9750278 TI - The seasonal variation of thermophilic campylobacters in beef cattle, dairy cattle and calves. AB - The epidemiology of clinical cases of campylobacter in temperate climates shows a striking seasonality. In the search for a seasonal environmental reservoir changes in the carriage rate and population size of campylobacters in bovine hosts with time have been measured. Most probable number (MPN) methodology was used to enumerate thermophilic campylobacters in samples taken from the small intestines of beef cattle at slaughter and the fresh faeces of four dairy herds and new-born calves. Statistical analyses revealed significant evidence for seasonal periodicity in the data from dairy herds (P = 0.044). Not only was there a departure from constancy within a 12-month interval but these data revealed a true seasonality, that is, the same periodicity in numbers from one year to the next. Each herd had two peaks per year, in approximately spring and autumn. Peaks coincided in herds on neighbouring farms but those on farms in the north preceded those on farms in the south by 2 and 1 months, respectively (P = 0.0057). Intestinal carriage by beef cattle at slaughter was 89.4% (n = 360) with an average MPN campylobacters per gram fresh weight (MPN gfw-1) of 6.1 x 10(2). Average MPN gfw-1 in faeces from the dairy herds and calves were 69.9 (S.D. 3) and 3.3 x 10(4) (S.D. 1.7 x 10(2)). There was no evidence of seasonal periodicity in the size of the campylobacter population in beef cattle at slaughter. Calves were campylobacter free at birth but became colonized with a few days. PMID- 9750279 TI - Synergistic antibacterial action of heat in combination with nisin and magainin II amide. AB - Lactobacillus plantarum has been exposed to mild heat at temperatures between 48 and 56 degrees C in combination with low concentrations of the lantobiotic nisin in different sequential set-ups. Exposure to heat and nisin caused synergistic reductions of Lact. Plantarum viability. Efficient antimicrobial action was dependent on the growth state of the culture as well as on levels and sequences of treatment applications. Listeria monocytogenes and Escherichia coli were treated at 55 degrees C in the presence of magainin II amide. Synergistic reductions in viable counts could be observed for L. monocytogenes and, after prolonged exposure, also for E. coli. the bacterial membrane could be identified by fluorometry and flow cytometry as an important target of applied treatment combinations. PMID- 9750280 TI - Analysis of biocide transport limitation in an artificial biofilm system. AB - An alginate gel bead artificial biofilm system was used to assay biofilm susceptibility to four biocides and to analyse the extent to which each agent penetrated the biofilm. Chlorine, glutaraldehyde, an isothiazolone, and a quaternary ammonium compound were tested on alginate-entrapped Enterobacter aerogenes in gel beads ranging from 1.8 to 6 mm in diameter. Gel-entrapped bacteria were less susceptible to all four antimicrobial agents than were planktonic micro-organisms. The degree of kill measured in artificial biofilm gel beads depended on the size of the gel bead and the cell density at which it was loaded. Disinfection efficacy decreased as gel bead radius or cell density increased. The manifest dependence of biofilm disinfection efficacy on the physical properties of the artificial biofilm (radius and cell density) suggests the impingement of transport limitation of biocide transport into the biofilm. A previously developed theory of biocide reaction and diffusion in biofilm was tested by calculating an appropriate Thiele modulus. In accordance with the theory, the efficacy of all four biocides decreased, albeit noisily, as the Thiele modulus exceeded 1. This result demonstrates that transport limitation can impact antimicrobial performance against biofilms not only of oxidizing biocides but also of nonoxidizing agents. PMID- 9750281 TI - Occurrence, diversity and pathogenicity of mesophilic Aeromonas in estuarine waters of the Italian coast of the Adriatic Sea. AB - A total of 208 strains of Aeromonas were isolated by monthly sampling from two estuaries (one provided with, and the other devoid of a waste-water treatment system) on the Italian coast of the Adriatic sea between September 1994 and August 1995. Biotyping at the species level allowed the identification of 96 strains (46%) as Aer. caviae, 46 (22%) as Aer. sobria, 33 (16%) as Aer. hydrophila and 25 (12%) as Aer. veronii. Eight strains (4%) were regarded as unnamed aeromonads. Aeromonas caviae was the most prevalent species in water with a high degree of pollution, while Aer. hydrophila strains were more commonly isolated from cleaner water. Aeromonas sobria and Aer. veronii were equally distributed in both estuaries. There was no correlation between temperature and numbers of aeromonads in either estuary. Using a biochemical fingerprinting method, strains were divided into similarity groups (PhP-types) based on their biochemical phenotypes. Several different PhP-types were found in each estuary, yielding a high diversity for these strains. However, some identical PhP-types were also found in both estuaries and at different times of the year, indicating that certain Aeromonas strains can survive more widely varying physico-chemical conditions. The production of toxins capable of causing cytoskeletal-dependent changes in the morphology of Chinese hamster ovary (CHO) cells was detected in 14 strains and appeared to be dependent on the season. PMID- 9750282 TI - Antilisterial activity of enterocin 81, a bacteriocin produced by Enterococcus faecium WHE 81 isolated from cheese. AB - Enterocin 81, a bacteriocin produced by Enterococcus faecium WHE 81 previously isolated from cheese, exhibited a very narrow spectrum of activity, which is mainly directed against enterococci and Listeria spp. including Listeria monocytogenes. Enterocin 81 activity, which was extremely rapid with maximal effect achieved within 30 min, could not be detected after treatment with various proteolytic enzymes. This activity was bactericidal in nature and induced an important efflux of intracellular material, which was visualized under electron microscopy as filaments coming out of L. monocytogenes cells. However, enterocin 81 did not display bacterial lysis on sensitive cells, as no changes in cell morphology were detected following the bactericidal action. Furthermore, this bacteriocin was shown to be equally active at pH values ranging from 4.0 to 8.0, which, along with the narrow activity spectrum, are two factors of paramount interest with regards to possible use of this bacteriocin in fermented foods. PMID- 9750283 TI - The diversity of lipases from psychrotrophic strains of Pseudomonas: a novel lipase from a highly lipolytic strain of Pseudomonas fluorescens. AB - Strains of Pseudomonas fluorescens and Ps. fragi are the predominant psychrotrophs found in raw milk and may cause spoilage due to the secretion of hydrolytic enzymes such as lipase and protease. The diversity of lipases has been examined in Pseudomonas isolates from raw milk which represent different taxonomic groups (phenons). Significant diversity was found using both DNA hybridization and immunoblotting techniques, which has implications for the development of a diagnostic test. The lipase-encoding gene (lipA) was cloned from one strain, C9, of Ps. fluorescens biovar V. In contrast to previously reported lipase sequences from Ps. fluorescens, the gene encodes a lipase of M(r) 33 kDa. Alignment of all known Pseudomonas and Burkholderia lipase amino acid sequences indicates the existence of two major groups, one of M(r) approximately 30 kDa comprising sequences from Ps. fragi, Ps. aeruginosa, Ps. fluorescens C9 and Burkholderia, and one of approximately 50 kDa comprising Ps. fluorescens lipases. The lipase from C9 does not contain a signal peptide and is presumed to be secreted via a signal peptide-independent pathway. The lipA gene of strain C9 was disrupted by insertional mutagenesis. The mutant retained its lipolytic phenotype, strongly suggesting the presence of a second lipase in this strain. PMID- 9750284 TI - Growth and enterotoxin production of Staphylococcus aureus during the manufacture and ripening of Camembert-type cheeses from raw goats' milk. AB - Tests were carried out to determine the effect of manufacturing procedures for a Camembert-type cheese from raw goats' milk on the growth and survival of Staphylococcus aureus organisms added to milk at the start of the process, and to study the possible presence of staphylococcal enterotoxin A in these cheeses. The initial staphylococcal counts were, respectively, 2, 3, 4, 5 and 6 log cfu ml-1. Cheese was prepared following the industrial specifications and ripened for 41 d. Detection of enterotoxins was done by the Vidas SET test and by an indirect double-sandwich ELISA technique using antienterotoxin monoclonal antibodies. Generally, numbers of microbes increased at a similar rate during manufacture in all cheeses until salting. During the ripening period, the aerobic plate count population and Staph. aureus levels remained stable and high. There was an approximately 1 log reduction of Staph. aureus in cheeses made with an initial inoculum of Staph. aureus greater than 10(3) cfu ml-1 at the end of the ripening period (41 d) compared with the count at 22 h. The level of staphylococcal enterotoxin A recovered varied from 1 to 3.2 ng g-1 of cheese made with an initial population of 10(3)-10(6) cfu ml-1. No trace of enterotoxin A was detected in cheeses made with the lowest Staph. aureus inoculum used in this study. PMID- 9750285 TI - Survival of Listeria monocytogenes in sea water and effect of exposure on thermal resistance. AB - Survival, recoverability and sublethal injury of two strains of Listeria monocytogenes, Scott A and an environmental strain KM, on exposure to sea water at 12.8 or 20.8 degrees C was determined using in situ diffusion chambers. Plate counts were used to assess recoverability and injury while 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) reduction was used to determine respiratory activity. T90 values (times for 10-fold decreases in numbers of recoverable cells) on non selective medium (trypticase soya agar with 0.6% yeast extract) at 12.8 and 20.8 degrees C were 61.7 and 69.2 h for L. monocytogenes Scott A, and 103.0 and 67.0 h for L. monocytogenes KM, respectively. On selective medium (Oxford agar), T90 values at 12.8 and 20.8 degrees C were 60.6 and 56.9 h for L. monocytogenes Scott A, and 83.0 and 65.9 h for L. monocytogenes KM, respectively. With Scott A, the percentage of sublethally injured cells at 12.8 and 20.8 degrees C was 1.7 and 17.7%, respectively, while for KM the values were 19.0 and 1.6%, respectively. The fraction of cells reducing CTC but which were not recoverable on plating progressively increased on exposure to sea water. Listeria monocytogenes KM challenged at 58 degrees C showed an apparent increase in heat resistance after exposure to sea water at 20.8 degrees C for 7 d (D58 = 2.64 min) compared with before exposure (D58 = 1.24). This increase in thermal resistance was not apparent at temperatures greater than 63 degrees C, and analysis of the best-fit regression lines fitted to the thermal data obtained from the two cell populations indicated that their thermal resistance was not significantly different (P > 0.05) over the temperature range tested (58-62 degrees C). PMID- 9750286 TI - Development and use of 16S rRNA gene targeted PCR primers for the identification of Escherichia coli cells in water. AB - The primary sequences of the V3 and V6 regions of the 16S rRNA gene of pathogenic and non-pathogenic strains of Escherichia coli were determined and compared with those obtained for a number of reference strains which belong to the family Enterobacteriaceae. Three oligonucleotide primers 16E1, 16E2 and 16E3 were designed and used in the polymerase chain reaction to identify specifically all E. coli isolates. When 16E1, 16E2 and 16E3 were used as primers for the identification of E. coli cells present in tap, underground and pond waters, as low as 1 cfu 100 ml-1 of water could be detected if an 8 h pre-culture step was performed prior to the PCR reaction. PMID- 9750287 TI - Changes in bacterial populations in the colon of pigs fed different sources of dietary fibre, and the development of swine dysentery after experimental infection. AB - Swine dysentery (SD) is a disease which can be controlled by feeding a diet low in dietary fibre. The influence of source and inclusion level of dietary fibre both on bacterial populations in the colon, and on subsequent development of SD in pigs experimentally infected with Serpulina hyodysenteriae was evaluated. In Experiment 1, pigs were fed a low-fibre diet based on cooked rice and a animal protein supplement, or the same diet containing added insoluble (iNSP, fed as oaten chaff) or soluble (sNSP, fed as guar gum) non-starch polysaccharides, resistant starch (RS), or a combination of the last two (sNSP/RS). In Experiment 2, different levels of RS were added to the diet. With the base rice diet and with the addition of iNSP, the total number of colonic bacteria was low, the Gram positive population predominated, S. hyodysenteriae did not colonize and SD did not develop. Synergistic bacteria (Fusobacterium necrophorum and Fus. nucleatum), which have been reported to facilitate colonization by S. hyodysenteriae, were found only among isolates from pigs fed the sNSP/RS diet, and these animals developed SD. Addition of RS to the diet increased total bacterial counts and stimulated growth of Gram-negative bacteria in the colon. In Experiment 1, this permitted colonization by S. hyodysenteriae, but not expression of SD. In contrast, in Experiment 2, this level of inclusion and two others allowed both colonization and development of SD. In conclusion, the addition of sNSP and/or RS to an otherwise protective rice-based diet generated changes in the large intestine microbiota which might have some influence on proliferation of S. hyodysenteriae and the development of SD. PMID- 9750288 TI - Bacteriophage and associated polysaccharide depolymerases--novel tools for study of bacterial biofilms. AB - Bacteriophage for three representative strains of Gram-negative biofilm bacteria have proved to be of widespread occurrence. Lytic bacteriophage have been isolated from local sewage for the bacterium 1.15, an exopolysaccharide (EPS) producing pseudomonad found originally as a component of biofilms in a local river, and for two Enterobacter agglomerans strains from industrial biofilms. Representative examples of all three bacteriophage possess a relatively low burst size and on solid media, exhibit very large plaques surrounded by a wide halo (5 20 mm) indicative of polysaccharide depolymerase action. The bacteriophage are thus similar to other viruses for EPS-producing bacteria in inducing the synthesis of enzymes degrading the polymers which occlude the bacterial cell surface. In each preparation, the polysaccharase activity was associated both with sedimented phage particles and with the supernate of bacterial lysates. The enzymes have been partially purified and used to prepare polysaccharide digests in which the major products from each polysaccharide are the presumed repeat units of the polymers or oligomers of these. The soluble phage enzymes each degrade their substrate by acting as endo-glycanohydrolases. The phage and their associated enzymes thus provide very useful highly specific tools for studies of biofilms incorporating the bacterial host strains. Their potential applications in studies on bacterial biofilms are discussed. PMID- 9750289 TI - Assessment of microbial involvement in the elevation of copper levels in drinking water. AB - A study of bacterial populations in metropolitan Adelaide domestic reticulation pipes was conducted to investigate a possible link between copper in drinking water and biofilms. Biofilm densities from cold water copper pipes at 10 sample sites were measured by viable cell counts. The range detected was from < 2 x 10(1) to 3.25 x 10(7) cfu cm-2. Five isolates were selected for further experiments as they represented a range of responses to solvated copper and relative tendency for adhesion on glass slides. Drinking water supplied to the Adelaide Hills is high in total organic carbon (TOC; 22.57 mg Cl-1) and has a negative Langelier Index (LI;-1.16), whereas Adelaide metropolitan water undergoes filtration and has both a lower TOC and LI (10.72 mg Cl-1, LI,-0.49). Copper coupons were exposed to biofilm isolates (24h), washed and resuspended in Adelaide metropolitan and Adelaide Hills water. Copper coupons not exposed to biofilm isolates were suspended in respective waters as a control. After 5 d of incubation, the copper content of Adelaide Hills water (4.71 +/- 0.87 mg Cu l-1), in which the copper coupons were suspended, consistently exceeded values obtained in the metropolitan Adelaide water (1.17 +/- 0.249 mg Cu l-1). The concentration of copper in the Adelaide Hills water was influenced by the bacterial species forming the biofilm on the coupon, with Agrobacterium sp. producing significantly higher levels of soluble copper than the control. The experiments reported here indicate that the suspended organic carbon, the aggressivity of the water and the biofilm may independently or synergistically increase the dissolution of copper from pipes into drinking water. PMID- 9750290 TI - Detection and characterization of a novel antibacterial substance produced by a Lactobacillus delbrueckii strain 1043. AB - A novel antibacterial substance produced by a strain isolated from Bulgarian yellow cheese was characterized. The producer strain was identified by molecular typing to belong to the species Lactobacillus delbrueckii, which is a rare producer of bacteriocins. The inhibitory agent was heat stable and active against lactic acid bacteria species and several food-borne pathogens: Listeria monocytogenes, Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Yersinia enterocolitica and Y. pseudotuberculosis. Its sensitivity to amylolitic enzymes and lipase suggested that a lipid and carbohydrate moiety could be important for the activity. The amino acid content of the purified bacteriocin was estimated to 29 amino acids. The bacteriocin was shown to be small (3.6-6 kDa) by three different methods: HPLC gel-filtration, SDS-PAGE and amino acid contents. PMID- 9750291 TI - Glucose-induced acid tolerance appearing at neutral pH in log-phase Escherichia coli and its reversal by cyclic AMP. AB - Escherichia coli shifted from broth at external pH (pH0) 7.0 to pH0 7.0 broth plus glucose rapidly induced marked acid tolerance which also appeared, albeit to a lesser extent, plus maltose, sucrose or lactose. Tolerance appeared without the medium pH becoming acidic. Tolerance was most substantial when glucose was added at pH0 7.0 but was also appreciable at pH0 7.5, 8.0 and 8.5. Induction of tolerance by glucose was markedly reduced by cyclic AMP and essentially abolished plus NaCl or sucrose; the induction process was also reduced but not fully inhibited by chloramphenicol, tetracycline and nalidixic acid. Glucose-induced organisms showed less acid damage to DNA and beta-galactosidase and it is likely that this is because glucose induces a new pH homeostatic mechanism which keeps internal pH close to neutrality at acidic pH0. In conclusion, it is clear that glucose induces a novel acid tolerance response in log-phase E. coli at pH0 7.0; it is now known that induction of this response involves the functioning of extracellular induction components including an extracellular induction protein. PMID- 9750292 TI - The genus Rhodococcus. PMID- 9750293 TI - Bactericidal activity of carvacrol towards the food-borne pathogen Bacillus cereus. AB - Carvacrol, a natural plant constituent occurring in oregano and thyme, was investigated for its bactericidal effect towards the food-borne pathogen Bacillus cereus. Carvacrol showed a dose-related growth inhibition of B. cereus. At concentration of 0.75 mmol l-1 and above, total inhibition of the growth was observed. Below this concentration, carvacrol extended the lag-phase, reduced the specific growth rate and reduced the maximum population density. Incubation for 40 min in the presence of 0.75-3 mmol l-1 carvacrol decreased the number of viable cells of B. cereus exponentially. Spores were found to be approximately 2 3 fold less sensitive to carvacrol than vegetative cells. Bacillus cereus cells showed reduced susceptibility towards carvacrol at pH 7.0 compared with different values between pH 4.5 and 8.5. The culture and exposure temperatures had a significant influence on the survival of vegetative cells. The highest death rate of cells was observed at an exposure temperature of 30 degrees C. Membrane fluidity was found to be an important factor influencing the bactericidal activity of carvacrol. PMID- 9750294 TI - The colonization of turkeys by thermophilic campylobacters. AB - The rate at which five broods of turkey chicks became colonized by thermophilic campylobacters was investigated. Day-old chicks were normally free of campylobacters on arrival on the farm with colonization beginning within 7 d. The carriage rate was 100% by day 14 in three of the broods and by day 21 in the other two. Higher carriage rates were obtained with enrichment procedures than with direct plating. Two broods were investigated over an extended interval for the number of campylobacters shed in their faeces. In brood A, Campylobacters increased from day l to day 14 concomitant with increase in the carriage rate and in the number of chicks with diarrhoea. By day 39, when the birds were sold to other farms, the excretion rate had reached 6 x 10(7) campylobacters g-1 fresh faeces. Brood B. was monitored over 91 d and showed peaks in Campylobacter numbers on days 19 and 75, corresponding to peaks in the number of diarrhoeic samples. The introduction of new birds into the brood resulted in an increase in the Campylobacter population and in the number of birds with diarrhoea. Campylobacter jejuni was the only species isolated but comprised several different biotypes. Analysis of the number of campylobacters at different sites along the gastrointestinal tract of mature turkeys at slaughter showed that numbers increased with distance from the beak and were highest in the caeca. PMID- 9750295 TI - Oxygen sensitivity of heated cells of Escherichia coli O157:H7. AB - Following defined heat treatments (55 degrees C for 100 min, 50 degrees C for 5 min, 61 degrees C for l min), a 6 decimal (6-D) reduction was obtained when cells of Escherichia coli O157:H7 were enumerated in aerobic growth medium. Part of this reduction (3-D) was due to thermal inactivation (as determined when cells were enumerated in anaerobic growth medium), and part (3-D) was due to the inability of sub-lethally heat-injured cells of E. coli O157:H7 to grow in the presence of oxygen. When held anaerobically, the injured cells regained their ability to grow in the presence of oxygen. Following heating at 59 degrees C for 5 min, repair took 4 h at 30 degrees C, 48 h at 20 degrees C, 95 h at 10 degrees C, but did not occur in 816 at 5 degrees C. Recovery from sub-lethal heat injury was not influenced by heat shock. These findings are relevant to the safety of minimally-heated foods. PMID- 9750296 TI - Microbial disinfection capacity of municipal solid waste (MSW) composting. AB - The disinfection capacity of a municipal solid waste (MSW) composting plant (Siloda) has been evaluated. In spring and summer, MSW was followed during the composting process from raw material to mature compost and long-term storage (1 year). Ascaris eggs, Salmonella, Shigella, total streptococci, faecal streptococci, total coliforms, faecal coliforms and Escherichia coli were studied. Disinfection was successful in terms of a decrease in faecal contamination indicators and disappearance of faecal pathogens. Faecal coliform concentration in raw waste reached 2.1 x 10(8) cfu g-1 dry weight in spring (CI 95%:5.2 x 10(7)-3.4 x 10(8)) and 7.2 x 10(8) cfu g-1 dry weight (1 x 10(8)-1.7 x 10(9)) in summer, and fell to less than 100 cfu g-1 dry weight within 20 d. Faecal streptococci concentrations reached 8.7 x 10(8) cfu g-1 dry weight (3.7 x 10(8)-1.3 x 10(9)) in spring and 2.0 x 10(9)cfu g-1 dry weight (5.6 x 10(8)-3.4 x 10(9)) in summer, and fell to 8.7 x 10(4) cfu g-1 dry weight (6.9 x 10(4)-1.0 x 10(5)). No seasonal pattern of contamination, mainly of animal origin, was observed. Microbiological quality of finished compost depends on the storage conditions. Therefore, the storage stage should be viewed as part of the composting process. Monitoring disinfection capacity of MSW composting needs to combine several microbial populations. PMID- 9750297 TI - Combined use of immobilized Candida stellata cells and Saccharomyces cerevisiae to improve the quality of wines. AB - Grape must fermentation by the combination of immobilized Candida stellata cells and Saccharomyces cerevisiae was carried out in order to enhance the analytical profiles of wine. Batch and continuous pre-treatment of must with immobilized C. stellata cells, followed by an inoculum of S. Cerevisiae, enhanced the analytical profiles of fermentates. The metabolic interactions between the two yeast species showed a positive influence on reducing sugars, acetaldehyde and acetoin metabolism. Sequential fermentation was the best combination for improving the analytical profiles of wine but caused a loss of viability and metabolic activity of beads by limiting their successive use. Continuous pre-treatment of must on the beads of C. stellata could be a more interesting modality to improve the quality of wines. This biotechnological process could be profitably used to produce specific and special wines. PMID- 9750299 TI - The occurrence of Aeromonas species in drinking water supplies of an area of the Dolomite Mountains, Italy. AB - A study was made of the occurrence of Aeromonas spp. in drinking water supplies in a mountain area in northeast Italy (the Dolomites). On account of its location, the water in question is exposed to a low level of pollution and systematic chemical disinfection is not necessary. Out of 7395 water samples analysed over a 3 year period, 1623 (21 x 95%) were found to be positive for Aeromonas, with levels ranging from 1 to 240 cfu 100 ml-1; 72 x 4% of the strains were identified as Aer. hydrophila, 14 x 7% as Aer. caviae and 12 x 9% as Aer. sobria. The percentage of recovery from surface water (approximately 40%) was found to be higher than that of ground water (springs: 24 x 9%; wells : 28x 6%). Aeromonas spp. were isolated from 21 x 7% of samples from the distribution network and showed no significant variations compared with water from reservoirs. There was no evidence, therefore, of after-growth in the distribution system. No correlation was found between the concentration of Aeromonas spp. and faecal indicator organisms. As the distribution of Aeromonas spp. was unrelated to anthropic pollution, it is believed that the search for these micro-organisms should be adopted as a further indicator of drinking water quality, especially in waters such as those in the present investigation not undergoing systematic purification treatment. PMID- 9750298 TI - Discharge of disinfected wastewater in recipient aquatic systems: fate of allochthonous bacterial and autochthonous protozoa populations. AB - The discharge of disinfected effluents affects the bacterivorous ability of protozoa and the effect depends on the disinfectant applied. Chlorine provokes a decrease in the number of protozoa and a delay in the bacterivorous ability. The discharge of ozonated and peracetic acid-treated wastewater provokes only an initial slight decrease in bacterivorous ability. No correlation was found between toxicity values detected using the Microtox assay and the effect of disinfected effluents on freshwater protozoa population. After the disinfection processes, recipient systems (fresh and marine water) have different effects on the survival of Escherichia coli populations discharged to them. The effect of the freshwater recipient system is less negative than the effect provoked by sea water, and the differences detected depend on the disinfection treatment applied. The wastewater bacterial population as a whole is able to grow after discharge of disinfected wastewater to receiving waters. However, in the absence of predation or competition, the recipient systems exert selection, with rod-shaped bacteria predominating. PMID- 9750300 TI - Salmonella in animal slurry can be destroyed by aeration at low temperatures. AB - Cattle and other animals infected by Salmonella can emit high numbers of these bacteria. To determine an effective means for reducing this bacterial group in animal slurry, samples were subjected to aeration in laboratory experiments and in farm-scale slurry tanks. A clear reduction in Salmonella levels was found in laboratory experiments at temperatures from 4 to 40 degrees C. Aeration in farm scale slurry tanks increased the temperature above the ambient temperatures (often less than 0 degrees C) to maxima ranging between 19 and 40 degrees C. Farm scale aeration results in similar reductions in Salmonella as those achieved in laboratory experiments. Thus, reductions, ranging from greater than 99% of the initial number to no detectable Salmonella, could be reached after 2-5 weeks using aeration processes with cattle slurries contaminated by Salm. infantis or pig slurry contaminated by Salm. typhimurium. These results suggest that farmers can control the spread of Salmonella from slurry to agricultural fields. The reduction mechanisms remain unknown, though the increase in pH (to 7 x 6-9 x 0) found in slurries after aeration might exert a decreasing effect on these bacteria. PMID- 9750301 TI - Presence of Helicobacter species DNA in Swedish water. AB - Municipal water, treated waste-water and well-water from all 25 counties in Sweden were analysed for the presence of Helicobacter spp. DNA. Bacteria were concentration by immunomagnetic separation. Culture, Gram staining and urease tests were performed before lysis of bacteria. Two polymerase chain reaction (PCR) assay with high sensitivity (adhesin and 16S rRNA) were followed by Helicobacter spp. specific hybridization. Nine of 24 private wells, three of 25 municipal tapwater and three of 25 wastewater samples were positive for both PCR assays. Positive municipal and waste-water samples were positive for counties. Non-specificity of PCR methods due to the presence of unknown bacteria within the genus helicobacter cannot be totally ruled out. Thus, the clinical significance of findings Helicobater spp. DNA in drinking water needs to be further evaluated. PMID- 9750302 TI - Bacteriocins inhibit glucose PEP:PTS activity in Listeria monocytogenes by induced efflux of intracellular metabolites. AB - Glucose transport by the phosphoenolpyruvate (PEP)-dependent phosphotransferase system (PTS) of listeria monocytogenes is inhibited by the bacteriocins, nisin, pediocin JD and leuconocin S. To investigate the mechanism of inhibition, PTS activity assays were performed with permeabilized, bacteriocin-treated L. monocytogens Scott A cells. In the presence of exogenous PEP, nisin stimulated the PTS while both pediocin JD and leuconocin S partially inhibited its activity. These results suggested that PTS enzymes were still active in bacteriocin-treated cells and the bacteriocin-induced PEP efflux may be a mechanism for inhibition of the PTS. To verify that PEP did efflux from bacteriocin-treated L. monocytogens Scott A cells, intracellular and extracellular PEP were measured by HPLC. All three bacteriocins induced efflux of PEP. Nisin, pediocin JD and leuconocin S also induced efflux of AMP, ADP and ATP. These studies indicate that bacteriocin inhibition of the glucose PEP:PTS in L. monocytogenes is due to efflux of intracellular metabolites, particularly PMID- 9750303 TI - A study of the growth kinetics of Yersinia enterocolitica serotype O:3 in pure and meat culture systems. AB - The growth kinetics of a virulence plasmid-bearing (P+) and a plasmid-cured (P-) strain of Yersinia enterocolitica serotype O:3 in pure and meat culture were investigated. Growth studies were carried out at 25 and 37 degrees C in supplemented phosphate-buffered saline, buffered peptone water, cefsulodin irgasan-novobiocin broth base or supplemented broth base (CIN). The lag phase durations and growth rates under these conditions were determined by linear regression analysis. In pure culture, under most sets of equivalent conditions, P+ and P- strains had similar lag phase durations. However, under one set of conditions, i.e. CIN broth at 37 degrees C, the lag phase duration of the P+ strain was significantly longer than P-. In all but the most selective medium, P+ strains had slower growth rates that P- strains at 37 degrees C, probably due to the increase metabolic burden entailed in the maintenance of the virulence plasmid. In the most selective medium, i.e. CIN broth, P+ strains grew significantly faster than P-. This finding suggests that possession of virulence plasmid confers an enhanced ability to grow in the presence of selective agents. In meat cultures, both strains had longer lag phase than in equivalent pure cultures, with longer lag phases noted at 37 than at 25 degrees C. No significant differences were observed between the length of lag phases of P+ and P strains in meat culture. Both strains of Y. enterocolitica displayed faster growth rates in meat cultures than in pure cultures, indicating that one of more components of meat enhanced the growth of this organism. The effects and interaction of incubation temperature, enrichment broth and meat on the growth kinetics of plasmid-bearing and plasmid-cured Y. enterocolitica strains are discussed. PMID- 9750304 TI - Phospholipids of Fusobacterium spp. AB - The aim of this study was to analyse individual polar lipid analogues, within each lipid family present, of fusobacteria using fast atom bombardment mass spectrometry (FAB-MS). Polar lipid extracts were prepared, washed and dried. Samples, dispersed in a matrix of m-nitrobenzyl alcohol, were analysed by negative ion FAB-MS using xenon as the reagent gas. Major anion peaks observed in the low mass region of mass/charge (m/z), 211, 221, 225, 227, 239, 241, 249, 251, 253, 255, 273, 277, 279, 281, 289 and 291, were consistent with the presence of C13:1, C14:3, C14:1, C14:0, C15:1, C15:0, C16:3, C16:2, C16:1, C16:0, unknown, C18:3, C18:2, C18:1, unknown and C19:3 carboxylate anions. In the high mass region, major anion peaks observed with m/z 644, 646, 648, 660, 662, 672, 673, 674, 686, 688, 689, 690, 698, 700, 701, 703, 714, 716, 717 and 719 were consistent with the presence of phosphatidylethanolamine (PE) (29:2), PE (29:1), PE (29:0), PE (30:1), PE (30:0), PE (31:2), first isotope of PE (31:2), PE (31:1), PE (32:2), PE (32:1), first isotope peak of PE (32:1), PE (30:0), PE (33:3), PE (33:2), phosphatidylglycerol (PG) (31:3), PG (31:2), PE (34:2), PE (34:1), PG (32:2) and PG (32:1). We conclude that FAB-MS can provide data on individual analogues of PE and PG from Fusobacterium spp. not readily obtained by other means. Furthermore, the phospholipid profile is diagnostic for the genus. PMID- 9750305 TI - Isolation of isoproturon-degrading bacteria from treated soil via three different routes. AB - Three different isolation routes (flask enrichment/flask degradation assay, flask enrichment/microplate degradation assay, MPN assay/microplate degradation assay) were used to obtain pure cultures of bacteria which degraded isoproturon (3-(4 isopropylphenyl)-1,1-dimethylurea) as sole carbon and nitrogen source in a mineral salts medium from a field soil treated with isoproturon in the laboratory. All three isolation routes were successful, but the microplate assay of degradation was more successful than the flask assay. Characterization of 36 isolates indicated that they formed 16 distinct phenotypes (10 Gram-positive phenotypes, six Gram-negative phenotypes) which are likely to represent distinct species. Low concentrations of the degradation product 3-(4- isopropylphenyl)-1 methylurea (IPPMU) were occasionally found in the culture solutions. When provided as the sole source of carbon and nitrogen, the monomethyl degradation product was itself rapidly degraded by several of the isolates. Some isolates were also able to use the demethylated degradation product 3-(4-isopropylphenyl) urea (IPPU) as sole source of carbon and nitrogen, although there was occasionally an extended lag-phase before rapid degradation commenced. One isolate was particularly active and degraded isoproturon, the monomethyl and demethylated degradation products of isoproturon, and demethylated the related phenylureas diuron and linuron. PMID- 9750306 TI - Identification of a gene encoding a methyl-accepting chemotaxis-like protein form Campylobacter coli and its use in a molecular typing scheme for campylobacters. AB - Using PCR amplification with degenerate primers, a gene (tlpA) form Campylobacter coli encoding a putative 63x0 kDa polypeptide which exhibited significant identity with bacterial methyl-accepting chemotaxis proteins (MCPs) was identified. A mutant containing an inactivated copy of the tlpA A gene showed a wild-type chemotactic response to all of the chemo-attractants tested. A DNA probe based on the Highly Conserved Domain (HCD) of TlpA revealed the presence of multiple copies of genes encoding MCP-like proteins in both Camp. coli and Camp. jejuni. The arrangement of restriction sites within, and proximal to, genes with homology to the HCD probe varied among strains, resulting in a high degree of polymorphism. It is demonstrated here that a DNA probe compromising the HCD region of MCP-like proteins can be used, in Southern hybridization-based assays, to provide novel information which allows the discrimination of individual strains of Camp. coli and Camp. jejuni. PMID- 9750307 TI - Tissue culture assays using Caco-2 cell line differentiate virulent from non virulent Listeria monocytogenes strains. AB - Within the group of Listeria sp., only L. monocytogenes is pathogenic for humans and numerous studies of L. monocytogenes strains have described non-virulent isolates. In this study, the potential value of two tissue culture assays (TCA) was analysed to ascertain the virulence properties of L. monocytogenes strains, initially typed for virulence using the immunocompromised mouse model (ICMM). The first assay assessed both the penetration into, and multiplication within, Caco-2 cells (PM assay): the second was a plaque-forming assay (PF assay). All the clinical isolates (nine strains) were virulent in both TCA. Conversely, all the non-pathogenic species (seven strains) were non-virulent in PM and PF assays. Compared with the virulence obtained in the ICMM with 29 Listeria strains, including 12 non-virulent L. monocytogenes strains, the sensitivity of both TCA was equal to1. Specificity was 0.89 and 0.84 for the PF and PM assays, respectively. However, a study of strains exhibiting virulence differences in three other in vivo virulence models showed that ICMM only detected highly virulent strains. The specificity of the PF test could, therefore, be higher, and close to that obtained by the enumeration of viable bacteria in the spleen of mice infected by subcutaneous injection in the footpad and by intravenous injection. Taken together, this study confirms the existence of low-virulence L. monocytogenes strains and shows that the virulence status of some non-clinical L. monocytogenes isolates depends on the virulence models used. The data suggest that the PF assay could be used as a primary test to evaluate the virulence of Listeria strains in order to reduce the cost of testing all strains in vivo. PMID- 9750308 TI - Applicability of a model for non-pathogenic Escherichia coli for predicting the growth of pathogenic Escherichia coli. AB - A model was developed for the temperature dependence of growth rate of a non pathogenic Escherichia coli strain. The suitability of that model for predicting the growth rate of pathogenic E. coli strains was assessed. Growth rates of pathogenic strains were found to be adequately described by the model. Model predictions were also found to describe sufficiently well-published growth rate data for non-pathogenic E. coli on mutton carcase surfaces and E. coli O157:H7 in ground roasted beef, milk, and on cantaloupes and water melons. In addition, E. coli O157:H7 was found to grow in the region of 44-45 degrees celsius. PMID- 9750309 TI - Preliminary description of biocidal (syringomycin) activity in fluorescent plant pathogenic Pseudomonas species. AB - Strains representing the fluorescent plant pathogenic Pseudomonas spp., Ps. agarici, Ps. asplenii, Ps. avellanae, Ps. beteli, Ps. caricapapayae, Ps. cichorii, Ps. corrugata, Ps. ficuserectae, Ps. flectens, Ps. fuscovaginae, Ps. marginalis, Ps. meliae, Ps. savastanoi, Ps. syringae, Ps. tolaasii and Ps. viridiflava were tested for biocidal activity using Aspergillus niger as assay organism. Inhibitory behaviour was found in strains of Ps. asplenii, Ps. blatchfordae, Ps. cichorii, Ps. corrugata, Ps. fuscovaginae, Ps. marginalis, Ps. marginalis pv. pastinacea, Ps. syringae pv. syringae, Ps. syringae pv. aptata, Ps. syringae pv. atrofaciens, Ps. syringae pv. lapsa, Ps. tolaasii, and strains of a Pseudomonas sp. pathogenic to Actinidia, in the Ps. savastanoi genomic sp. Antifungal activity could be identified with the production of members of the syringomycin family of toxins by strains in Ps. syringae, Ps. asplenii and Ps. fuscovaginae. These toxin reactions support suggestions made elsewhere of the synonym of the latter two species. In a preliminary characterization using tests for stability to heat, protease, acid and alkaline treatments, unknown toxins consistent with syringomycin-like toxins the strains from Actinidia species. The toxins from Ps. cichorii and from Ps. corrugata differed in their reactions from all other agents. Pseudomonas tolaasii produces the antifungal compound tolaasin. The white line reaction with Ps. reactions, a test for tolaasin production by strains of Ps. tolaasii, was confirmed as specific for this compound. Some of these low molecular weight toxins may be produced by some of these plant pathogenic strains. PMID- 9750310 TI - Growth of a human intestinal Desulfovibrio desulfuricans in continuous cultures containing defined populations of saccharolytic and amino acid fermenting bacteria. AB - Ecological and physiological effects of the sulphate-reducing bacterium (SRB) Desulfovibrio desulfuricans on other intestinal organisms were investigated in anaerobic chemostats (dilution rate approximately 0.2 h-1). Reproducible defined bacterial communities were used in these experiments, comprising 14 different saccharolytic and amino acid fermenting species: Bifidobacterium longum, Bif. adolescentis, Bif. pseudolongum, Bif. infantis, Bacteroides thetaiotaomicron, Bact. vulgatus, Lactobacillus acidophilus, Enterococcus faecalis, Ent. faecium, Escherichia coli, Clostridium perfringens, Cl. butyricum, Cl. innocuum, Cl. bifermentans. Lactobacillus and Cl. bifermentans populations never rose above minimum detection limits (log10 2.0 and 4.0, respectively) under the experimental conditions employed in these studies. Inclusion of Des. desulfuricans in bacterial cultures (c. log10 8.4 viable cells ml-1) resulted in marked reductions (i.e. greater than 1 log) in planktonic cell population densities of several species, particularly Bif. longum, Cl. perfringens and Bif. pseudolongum. The two bacteroides species were unaffected by Des. desulfuricans, while numbers of Cl. butyricum increased. Extensive wall growth developed in the SRB culture, consisting mainly of Des. desulfuricans (log10 9.2 viable cells ml-1), Bact. thetaiotaomicron and Bact. vulgatus, with lesser numbers of facultative anaerobes, Cl. perfringens and Bif. longum. Wall growth was associated with a reduction in planktonic cell mass and increased acid production by the cultures. Chemotaxonomic study of chemostat microbiotas, on the basis of cellular fatty acid methyl ester (FAME) analyses, showed the existence of characteristic bacteroides (C15) and bifidobacterial (C18) markers, but desulfovibrio markers (i C15:0, C16:0, i-C17:1) could be identified. The metabolic activities of saccharolytic organisms were altered in the SRB chemostat, including synthesis of a number of hydrolytic enzymes involved in carbohydrate breakdown, such as alpha galactosidase, alpha-glucosidase and beta-galactosidase, together with several mucinolytic enzymes. High concentrations of sulphide (8.2 mmol 1-1) were detected in the SRB chemostat, suggesting that this metabolite may have been inhibitory to some species. Saccharolytic organisms growing in the SRB fermenter utilized more starch, but less galactose-containing polymers, which correlated with the observed glycosidase activities. Profound differences were also recorded with respect to fermentation product formation in the chemostats, where a major switch to acetate production occurred in the SRB culture, with concomitant reductions in propionate, butyrate and lactate, which is an important electron donor for desulfovibrios. PMID- 9750311 TI - Analysis of coccal cell formation by Campylobacter jejuni using continuous culture techniques, and the importance of oxidative stress. AB - Campylobacter jejuni is an important human gastrointestinal pathogen. In hostile environments it may adapt its physiology to prolong survival, potentially including the adoption of a viable, non-culturable form and a change to coccal cell morphology. By independently controlling the individual parameters of continuous cultures of Camp. jejuni (e.g. pH, nutrient limitation, growth rate, etc.), coccal cell formation was shown to be elicited only be high oxygen tension in conjunction with reduced carbon concentration. Electron microscopy revealed degradative changes in these cells. This occurred as a transient response over 48 h coincident with a large reduction in maximum growth rate and viable count. Kinetic analysis of the biomass reduction of the cultures demonstrated that significant underlying growth was maintained, with the subsequent selection of a more oxygen-resistant population of cells and reversion to spiral morphology. Coccal cells appear to be predominantly a degenerate form of Camp. jejuni resulting from oxidative damage. While some of these coccal cells may recover, the more interesting population of cells is probably that which retains, or regains, spiral morphology during adaptation to oxidative stress. PMID- 9750312 TI - Anomaly detection: eye movement patterns. AB - The symptom of a garden path in sentence processing is an important anomaly in the input string. This anomaly signals to the parser that an error has occurred, and provides cues for how to repair it. Anomaly detection is thus an important aspect of sentence processing. In the present study, we investigated how the parser responds to unambiguous sentences that contain syntactic anomalies and pragmatic anomalies, examining records of eye movement during reading. While sensitivity to the two kinds of anomaly was very rapid and essentially simultaneous, qualitative differences existed in the patterns of first-pass reading times and eye regressions. The results are compatible with the proposal that syntactic information and pragmatic information are used differently in garden-path recovery. PMID- 9750313 TI - Relatives children say. AB - In an experiment designed to elicit restrictive relatives clauses, 28 children ranging in age from 2; 2 to 3; 10 provided a corpus of communicatively appropriate relative clauses. In evaluating this corpus, we found that most children produced mostly adult relative clauses most of the time. Detailed study of these utterances uncovered a few error patterns, which we analyzed in light of several considerations (e.g. the overall frequency of an error type, its distribution across children and items, its relation to the construction under study, and the similarity of the error to what children do elsewhere). Only one error pattern, namely some children's production of inappropriate relative pronouns, is argued to reflect a systematic feature of language development. We conclude that children's ability to represent the syntactic structure of the embedded clause is on target very early. PMID- 9750314 TI - Physical mapping of the conjugative plasmid pMB1-1 of Enterococcus faecalis. AB - The sexpheromone system of Enterococcus faecalis is a form of bacterial conjugation that plays an important role in the horizontal dissemination of genes. The ecological significance of 'sexual' plasmids is to permit a rapid mobilization of genes of interest for the species (e.g. those encoding haemolysins, bacteriocins or antibiotic resistance). The physical mapping of pMB1 1, a conjugative plasmid of Ent. faecalis that responds to cCF10 pheromone, has been undertaken. By means of hybridization with conserved sequences of pCF10 plasmid, the regions harbouring the genes responsible for the pheromone inhibitor and the aggregation and exclusion proteins of this plasmid have been identified. The results demonstrated that plasmids pMB1-1 and pCF10 only show homology in the region involved in the conjugative response, suggesting that this region may be transferred in an independent way to that of the rest of the plasmid. PMID- 9750315 TI - Haemagglutination and glycolipid-binding activities of Lactobacillus reuteri. AB - The carbohydrate-binding activity of Lactobacillus reuteri was studied by haemagglutination (HA), HA inhibition and thin layer chromatography (TLC) overlay assays. Three of the six Lact. reuteri strains examined showed HA activity. Two strains (JCM1081 and JCM1112T) agglutinated neuraminidase-treated, but not untreated, erythrocytes. Strain JCM2762 agglutinated both treated and untreated erythrocytes. The HA activity of JCM 1081 was inhibited by galactose, lactose, methyl beta-galactoside and asialoglycophorin A. Among 12 glycosphingolipids, TLC overlay assay showed that JCM1081 strongly bound to asialo-GM1. These results indicated that JCM1081 bound to the beta-galactosyl residues of the non-reducing terminal of sugar chains of glycoconjugates. The carbohydrate-binding ability of JCM1081 may be responsible for the adhesion of this strain to the mucosal surface of the intestine. PMID- 9750316 TI - Comparative antibacterial effects of novel Pelargonium essential oils and solvent extracts. AB - The scented leaves of a number of Pelargonium (Geraniaceae) species and cultivars were extracted using steam distillation, petroleum spirit and methanol. The extracts were assessed for their antibacterial activity in vitro against Staphylococcus aureus, Proteus vulgaris, Bacillus cereus and Staph. epidermidis. The results indicated substantial antibacterial activity and suggested that Pelargonium essential oils could be used as novel antibacterial agents. The methanolic and petroleum spirit extracts were more potent antibacterial agents than the steam-distilled volatile samples. The results suggest that Pelargonium essential oils and solvent extracts could be used as novel food or cosmetic antimicrobial agents. PMID- 9750317 TI - Cross-species induction and enhancement of antimicrobial activity produced by epibiotic bacteria from marine algae and invertebrates, after exposure to terrestrial bacteria. AB - Antibiotic producing marine bacteria isolated from surfaces of the marine alga Fucus vesiculosus and the nudibranch Archidoris pseudoargus were exposed to live cells of Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli and heat killed cells of Staph. aureus. Twelve out of the 16 marine strains tested showed enhanced antimicrobial activity towards Staph. aureus, E. coli and Ps. aeruginosa following this exposure. Three out of seven strains tested showed enhanced antimicrobial activity when exposed to Ps. aeruginosa and three out of seven strains showed enhanced antimicrobial activity when exposed to E. coli. These results suggest that production of antimicrobial compounds by marine bacteria can be induced by the presence of terrestrial bacteria. This appears to be the first example of cross-species induction and enhancement of antimicrobial activity in marine bacteria and has important implications for the design of antibiotic screening assays and for an understanding of microbial competition in the environment. PMID- 9750318 TI - Direct inoculation into media containing bile salts and antibiotics is unsuitable for the detection of acid/salt stressed Escherichia coli O157:H7. AB - The efficiency of selective enrichment broths for the recovery of low numbers of acid/salt stressed Escherichia coli O157:H7 was determined. Stressed cultures were diluted to low levels and recovered in tryptone soya broth with added bile salts, to make modified tryptone soya broth, and buffered peptone water with various combinations of antibiotic supplementation including novobiocin, acriflavine and a mixture of vancomycin, cefsulodin and cefixime (VCC) at 37 degrees C and 42 degrees C. Significantly fewer stressed cells, in some cases as little as 0.3% of the starting population, were recovered by all the selective enrichment broths containing bile salts or VCC antibiotics compared to the nonselective controls. The use of such enrichments to recover low numbers of stressed E. coli O157:H7 may result in failure to detect the organism. Parallels with salmonella methodology are made and the need for a non-selective pre enrichment stage in E. coli O157:H7 methods discussed. PMID- 9750319 TI - Restriction enzyme analysis of randomly amplified polymorphic DNA amplicons of Salmonella enterica ser. Enteritidis PT4 and Typhimurium DT104. AB - Salmonella enterica serotype Enteritidis PT4 and Typhimurium DT104 isolates were characterized using a random amplification of polymorphic DNA (RAPD) protocol found previously to be highly discriminatory for isolates of Salmonella. Profiles generated with a single primer 1254, and independently 1283, successfully characterized an outbreak strain of Enteritidis PT4 but could not differentiate epidemiologically unrelated strains of Enteritidis PT4 from the outbreak strains. Primer 1254 differentiated one strain, and 1283 two strains of Typhimurium DT104 previously undifferentiated on the basis of biochemical and physical properties. Subsequent analysis using a combination of RAPD and restriction enzyme analysis could not provide additional differentiation of Enteritidis PT4 and Typhimurium DT104 isolates but did, however, exhibit the potential to be a useful combination of molecular techniques. PMID- 9750320 TI - Polymerase chain reaction for rapid detection of Campylobacter jejuni in artificially contaminated foods. AB - Campylobacter jejuni was inoculated into a range of raw and ready-to-eat foods. These food samples were used as a test source for detection of this bacterium by the polymerase chain reaction (PCR). Specific detection of Camp. jejuni, as indicated by a PCR product of 159 bp, was possible with all pre-cooked deli sliced and raw poultry products tested. All vegetables tested were compatible with the PCR assay. Cantaloupe, kiwi and pineapple tested positive while strawberries, watermelon, grapes and apples tested negative. By using a nested PCR assay that yields a single band at 122 bp, positive results were obtained with watermelon and grapes while the apple and strawberries continued to give a negative reaction. These rapid and specific assays for Camp. jejuni are compatible with most foods in insuring the safety of the food product. PMID- 9750321 TI - A RAPD-PCR method for large-scale typing of Bacillus cereus. AB - A robust RAPD-PCR procedure for large-scale typing of Bacillus cereus was developed. It is based on a simple DNA preparation, involving only freezing and boiling of cells in water with active carbon. By using a computerized system for data collection and processing, an efficient system for handling RAPD patterns for large-scale investigations was achieved. The procedure was highly discriminatory for Bacillus cereus strains and was found to give reproducible classification of RAPD fingerprints for five reference strains. PMID- 9750322 TI - Acid production from lactulose by dental plaque bacteria. AB - Representative strains of oral streptococci, lactobacilli and bifidobacteria were incubated overnight with lactulose or other carbohydrates and the final pH recorded. Most bacteria tested were able to metabolize lactulose with the exception of strains of Streptococcus salivarius, Lactobacillus acidophilus and Lact. fermentum. Streptococcus mutans produced most acid overnight but the initial rate of acid production from lactulose by uninduced cultures was very low. Plaque pH was monitored in 12 volunteers following rinsing the mouth with lactulose, sucrose or sorbitol or Lactulose BP. These studies in vivo showed both lactulose and Lactulose BP to exhibit low acidogenic potential. Thus, although plaque bacteria are capable of fermenting lactulose, the results suggest that lactulose is likely to pose a small acidogenic challenge to teeth under normal conditions of use. PMID- 9750323 TI - Occurrence of bacteriocin production among environmental enterococci. AB - The occurrence of enteroccoci in cattle dung water from the basins of 25 cattle farms of 15 northeastern Slovakia districts was screened as well as bacteriocin production among selective enterococcal isolates. The average total count of enterococci detected reached 5.0 x 10(3) cfu ml-1. Enterococcus faecium was the predominant species (25%) followed by Ent. casseliflavus (19.2%), Ent. faecalis (9.6%), Ent. avium and Ent. durans (1.9%). The antagonistic activity of isolates showed a mainly antilisterial effect. Enterococcus faecalis V24 strain produced a heat stable, largely hydrophobic antimicrobial substance with best production in the range pH 4 to 7 and with a strong inhibitory effect even against Gram negative bacteria. PMID- 9750324 TI - Acid and bile tolerance of Lactobacillus isolated from chicken intestine. AB - Twelve Lactobacillus strains isolated from chicken intestine were used to investigate acid and bile tolerance in vitro. Ten out of the 12 strains were slightly affected by 0.3% bile salts, showing a delay of growth (d) of 0.6-37.2 min compared with growth in control cultures. Two strains were not affected by the bile salts. Of the 12 strains, seven could be arbitrarily classified as resistant (d < 15 min) and five as tolerant (15 min < d < or = 40 min). Lactobacillus strains from the caecum showed better tolerance to acid than those from the ileum. Generally, the survival of the ileal strains was very low at pH 1.0 and 2.0, and moderate at pH 3.0. In contrast, caecal Lactobacillus strains could survive at pH 1.0 for up to 2 h of incubation; growth was moderate at pH 2.0 and good at pH 3.0 and 4.0. PMID- 9750325 TI - Light inactivation of food-related pathogenic bacteria using a pulsed power source. AB - The effects of high intensity light emissions, produced by a novel pulsed power energization technique (PPET), on the survival of bacterial populations of verocytotoxigenic Escherichia coli (serotype 0157:H7) and Listeria monocytogenes (serotype 4b) were investigated. Using this PPET approach, many megawatts (MW) of peak electrical power were dissipated in the light source in an extremely short energization time (about 1 microsecond). The light source was subjected to electric field levels greater than could be achieved under conventional continuous operation, which led to a greater production of the shorter bacteriocidal wavelengths of light. In the exposure experiments, pre-determined bacterial populations were spread onto the surface of Trypone Soya Yeast Extract Agar and were then treated to a series of light pulses (spectral range of 200-530 nm) with an exposure time ranging from 1 to 512 microseconds. While results showed that as few as 64 light pulses of 1 microsecond duration were required to reduce E. coli 0157:H7 populations by 99.9% and Listeria populations by 99%, the greater the number of light pulses the larger the reduction in cell numbers (P < 0.01). Cell populations of E. coli 0157:H7 and Listeria were reduced by as much as 6 and 7 log10 orders at the upper exposure level of 512 microseconds, respectively. Survival data revealed that E. coli 0157:H7 was less resistant to the lethal effects of radiation (P < 0.01). These studies have shown that pulsed light emissions can significantly reduce populations of E. coli 0157:H7 and L. monocytogenes on exposed surfaces with exposure times which are 4-6 orders of magnitude lower than those required using continuous u.v. light sources. PMID- 9750326 TI - Escherichia coli O157 serology: false-positive ELISA results caused by human antibodies binding to bovine serum albumin. AB - An analysis of farm workers and rural dwellers for serum antibodies to the lipopolysaccharide (LPS) of Escherichia coli O157 detected sera with antibodies binding to bovine serum albumin (BSA) by ELISA. These antibodies were not specific for BSA when examined by immunoblotting, and the ELISA values were reduced to a background level when plates were blocked with normal rabbit serum. PMID- 9750327 TI - Effect of permeabilizing agents on antibacterial activity against a simple Pseudomonas aeruginosa biofilm. AB - A simple Pseudomonas aeruginosa G48 biofilm on stainless steel discs provided a useful primary screen of potentiating effects of various permeabilizing agents on antibacterial agents. Experiments with Ps. aeruginosa suspensions could not be used to predict the effects of biocides and permeabilizers on biofilms. Although antibacterial activity against biofilms was less than demonstrated in suspension tests, potentiation by some permeabilizers was still observed. PMID- 9750328 TI - Two restriction endonucleases in Selenomonas ruminantium subsp. lactilytica. AB - Crude protein extract from a recently isolated ruminal bacterium identified as Selenomonas ruminantium subsp. lactilytica specifically cleaved DNA. This ability was due to the presence of two site-specific restriction endonucleases. Sr/I, a NaeI schizomer, recognizes the 5'-GCCGGC-3' sequence. Sr/II, a NsiI schizomer, recognizes 5'-ATGCAT-3'. PMID- 9750329 TI - Degradation of 2,4,6-trichlorophenol by a specialized organism and by indigenous soil microflora: bioaugmentation and self-remediability for soil restoration. AB - A selected mixed culture and a strain of Alcaligenes eutrophus TCP were able to totally degrade 2,4,6,-TCP with stoichiometric release of Cl-. In cultures of Alc. eutrophus TCP, a dioxygenated dichlorinated metabolite was detected after 48 h of incubation. Experiments conducted with soil microcosms gave evidence that: the degradative process had a biotic nature and was accompanied by microbial growth; the soil used presented an intrinsic degradative capacity versus 2,4,6 TCP; the specialized organism used as inoculum was effective in degrading 2,4,6 TCP in a short time. These results could be utilized for the adoption of appropriate remediation techniques for contaminated soil. PMID- 9750330 TI - Anti-MRSA activity of sophoraflavanone G and synergism with other antibacterial agents. AB - Anti-MRSA activity of sophoraflavanone G (SFG) and synergism between SFG and antibacterial agents against MRSA (methicillin-resistant Staphylococcus aureus) were evaluated by means of Minimal Inhibitory Concentrations (MIC). The MICs of SFG against 27 strains of MRSA ranged from 3.13 to 6.25 micrograms ml-1. Synergism between SFG and vancomycin hydrochloride (VCM) or fosfomycin (FOM) was observed (the fraction inhibitory concentration (FIC) indices were 0.16 and 0.48), while partial synergism was admitted between SFG and other antibacterial agents such as methicillin (DMPPC), cefzonam (CZON), gentamicin (GM), minocycline (MINO) and levofloxacin (LVFX) (the FIC indices were 0.71, 0.73, 0.69, 0.65 and 0.58, respectively). These findings suggest that SFG in combination with VCM or FOM may be useful in controlling MRSA infections. PMID- 9750331 TI - Changes in the strength of attachment of micro-organisms to surfaces following treatment with disinfectants and cleansing agents. AB - Suspensions of Pseudomonas aeruginosa and Staphylococcus epidermidis, and biofilms established (16 h) on submerged glass and stainless steel (216 2B) coupons, were exposed to sodium hypochlorite (0.02% or 0.015% w/v), Dodigen (0.0015% w/v or 0.0006% w/v), sodium dodecylsulphate (6% w/v or 0.1% w/v) and Tween-80 (6% w/v) for 5 min at 20 degrees C. Survival was assessed by viable counts and blot succession. Biofilm bacteria were significantly less susceptible to these biocides than were planktonic cells, but their attachment to the surfaces was loosened by such treatments. Treatment with the non-ionic surfactant, Tween-80, however, strengthened the attachment of Staph. epidermidis to stainless steel. Such effects on attachment strength, which are species and surface dependent, have profound implications on post-treatment cleansing and possible re-contamination of product in clean-in-place (CIP) systems. PMID- 9750332 TI - Comparison of conventional culture and PCR methods for the detection of Legionella pneumophila in water. AB - A comparative assessment of conventional culture and nucleic acid techniques in the detection of Legionella pneumophila in seeded tap water samples was performed, using bacterial concentrations ranging from 994 to 0.015 cfu ml-1. Different filtration and centrifugation protocols were evaluated. The results permitted the development of a tentative algorithm for the detection of legionellae in tap water. Samples should first be analysed using PCR methods. In the event of quantitative data and bacterial strains for epidemiologic typing being required, the same sample, or a greater volume of the sample, if positive with PCR, can be re-tested by filtration through polycarbonate membranes followed by plating a homogenate of the filter. If samples are found to be negative with PCR, they can be re-analysed in greater volumes by filtration through polycarbonate membranes followed by direct placing of the filter on culture media, to allow detection of very low numbers of bacteria. This protocol should be validated in the field before it can be routinely implemented. PMID- 9750333 TI - Simple and direct detection of Staphylococcus aureus in milk by a tube coagulase test. AB - A tube coagulase test (TCT) is described as a simple and non-expensive system for detection of Staphylococcus aureus directly in milk. The procedure is characterized by mixing milk samples with rabbit citrate plasma followed by incubation at 37 degrees C for clot formation. The tube coagulase test demonstrated 91.5% accuracy, 88.5% sensitivity and 100% specificity for the direct recognition of Staph. aureus in milk samples from quarters with subclinical mastitis, when compared with plating of milk on blood agar. The TCT has the potential to detect other coagulase positive staphylococci in milk. It is concluded that TCT may be of use to veterinary practitioners with limited laboratory facilities, or to dairy farmers as a simple diagnostic test on site. PMID- 9750334 TI - The use of alkaline phosphatase-labelled oligonucleotide probes as culture confirmation reagents for identification of commercially important bacteria. AB - A range of rRNA-targeted alkaline phosphatase labelled oligonucleotide probes was tested for use as culture confirmation reagents for the rapid identification of micro-organisms. The probes were specific to clinically important bacteria (Helicobacter pylori and Mycobacterium tuberculosis), fish and shellfish pathogens (Renibacterium salmoninarum and Vibrio vulnificus), food spoilage bacteria (Listeria spp. and L. monocytogenes), for bacteria of biotechnological importance (Streptomyces spp.) and for bacteria associated with the oil industry (Sulphate-reducing bacteria, SRB). A universal bacterial probe and a eukaryotic probe were included in the study as positive and negative controls, respectively. A total of 93 bacterial strains was screened. With the exception of a large number of cross-reactions of the SRB probe (specificity value of 29.4%) and a single cross-reaction of the R. salmoninarum probe (specificity value of 97.7%), dot blot analysis indicated that each probe hybridized 100% specifically to the organisms tested. A simple culture confirmation method was then developed using these probes to enable the identification of bacterial colonies using a simple hybridization procedure. PMID- 9750335 TI - Regulation of superoxide dismutase synthesis in Candida albicans. AB - The synthesis of superoxide dismutase [SOD: EC 1.15.1.1] in response to various cultural conditions was examined in Candida albicans, an opportunistic yeast which causes candidiasis in immunosuppressed patients. SOD plays an important role in protecting cells from teh oxidative damage of superoxide radicals. Maximum SOD activity was found after 72 hrs of yeast growth. The optimum pH and temperature for the SOD activity were 7 and 40 degrees C, respectively. The major SOD activity was found in the cytosol fraction and the level of extracellular SOD was very low. The enzyme was stimulated to varying degrees by cholic acid, procaine and tocopherol. On the basis of inhibitor studies and other enzyme properties, the isolated enzyme from C. albicans is identified as a copper and zinc superoxide dismutase. PMID- 9750336 TI - Fungemia in cancer patients in Brazil: predominance of non-albicans species. AB - The objective of this study was to characterize the epidemiology of candidemia in cancer patients in the city of Rio de Janeiro, Brazil. An 18-month survey of fungemia in patients with cancer was undertaken in three Hospitals in Rio de Janeiro. Forty-three episodes of candidemia were identified in 43 patients, 43 of which were episodes of candidemia; in ten case the strains were not available for further identification of species and were excluded from this analysis. The overall distribution of fungi causing fungemia was: Candida albicans (5), Candida tropicalis (16), Candida parapsilosis (6), Candida guilliermondii (4), Candida lusitaniae (1) and Candida stellatoidea (1). Antifungal prophylaxis had been administered before the episode of fungemia in only six patients (18.2%): oral itraconazole in three patients and oral nistatin, low dose intravenous amphotericin B and oral fluconazole in one patient each. There was no difference in the presence of risk factors, clinical characteristics or in the outcome between albicans and non-albicans species, nor between Candida tropicalis and other non-albicans species. There was a clear predominance of non-albicans species, regardless of the underlying disease, antifungal prophylaxis or the presence of neutropenia. PMID- 9750337 TI - Lymphangitic sporotrichosis: an uncommon bilateral localization. AB - Sporotrichosis is a mycotic disease caused by cutaneous inoculation of the dimorphic fungus Sporothrix schenckii. The primary lesion can spread and often develop a unilateral lymphocutaneous lesions or, rarely, disseminated disease. We report a lymphangitic sporotrichosis case with ulcerated erythematous nodules distributed bilaterally on the posterior and medical aspect of the both legs, probably due to multiple inoculations. The treatment with oral potassium iodide was satisfactory. PMID- 9750338 TI - In vitro activity of a new triazole antifungal agent, Sch 56592, against clinical isolates of filamentous fungi. AB - Sch 56592 is a new triazole derivative that possesses potent, broad-spectrum antifungal activity. We evaluated the in vitro activity of Sch 56592 compared with that of itraconazole, amphotericin B and 5-fluorocytosine against 51 clinical isolates of filamentous fungi, including Aspergillus flavus (10), A. fumigatus (12), Fusarium spp. (13), Rhizopus spp. (6), Pseudallescheria boydii (5), and one isolate each of Acremonium spp., A. niger, A. terreus, Paecilomyces spp., and Trichoderma spp. In vitro susceptibility testing was performed using the microdilution broth method outlined in the NCCLS 27-A document. Sch 56592 was highly active against A. flavus (MIC90, 0.25 micrograms/ml), A. fumigatus (MIC90, 0.12 micrograms/ml), P. boydii (MIC50, 1 microgram/ml) and Rhizopus spp (MIC50, 1 microgram/ml). By comparison with itraconazole, Sch 56592 was four-to eight-fold more active against isolates of Aspergillus and both compounds showed equipotent in vitro activity against P. boydii and Rhizopus spp. Sch 56592 was four- to 16 fold more active than amphotericin B against Aspergillus spp. and P. boydii and both antifungal drugs displayed similar activity against Rhizopus spp. Overall, Sch 56592 showed good in vitro activity against all isolates tested (MIC, < or = 2 micrograms/ml) except isolates of Fusarium (MIC range, (1-->4 micrograms/ml). On the basis of these data Sch 56592 has promising activity against Aspergillus spp. and other species of filamentous fungi that are likely to be encountered clinically. Additional in vitro and in vivo studies are warranted. PMID- 9750340 TI - Identification of a DNA fragment encoding a putative membrane transporter protein expressed in Sporothrix schenckii. AB - We have isolated two DNA fragments from Sporothrix schenckii. These fragments were nonspecifically amplified from the whole-cell DNA using polymerase chain reaction (PCR) primers originally designed in the gene encoding heat shock protein 70 of Mycobacterium leprae. Reverse transcription PCR demonstrated that the smaller (sp-2) fragment was expressed, and a database search indicated that the deduced amino acids sequence from the sp-2 fragment contained a region homologous to the conserved sequence of the membrane transporter protein family. This is the first report of partial cloning of the gene encoding the putative membrane transporter in S. schenckii. PMID- 9750339 TI - Study of pulmonary experimental paracoccidioidomycosis by analysis of bronchoalveolar lavage cells: resistant vs. susceptible mice. AB - Adult Swiss (susceptible) and BALB/c (non-susceptible) mice were inoculated by the intravenous route with 1 x 10(6) yeast cells of Paracoccidioides brasiliensis, strain 18. Immunologic parameters, histopathology and features of the bronchoalveolar lavage (BAL) were evaluated at week 2, 4, 8 and 16 post infection. The pulmonary infection was progressive in Swiss mice and regressive in Balb/c mice. The numbers of total cells, lymphocytes and polymorphonuclear neutrophils increased in BAL, as well as the percentages of giant cells, and CD4 and CD8 positive cells. The ultrastructural study of BAL cells revealed a predominance of macrophages and a frequency of 13.2% of type II pneumocytes. As the infection progressed, the number of fungal cells and spreading macrophages, as well as the stimulated release of H2O2 by macrophages, increased. The animals exhibited an exacerbation of the humoral immune response and a depression of cellular immunity during the infection. There was a good correlation between the intensity and the pattern of the pulmonary histopathology and the cellular findings in the BAL. The present model reproduces some anatomoclinical patterns of the human disease and shows that BAL may be a useful tool in monitoring the pulmonary infection caused by P. brasiliensis. PMID- 9750341 TI - Activity of hydrolytic enzymes of Candida albicans strains isolated from patients with periodontal and membrane mucosae of oral cavity diseases. AB - Fungi are elements of the ontocenosis of the oral cavity and causal factors of inflammatory lesions in its mucous membrane. The objective of the study was to find differences in the activity of hydrolytic enzymes of Candida albicans isolated from patients with diseases of the periodontium and mucous membrane of the oral cavity. Of 235 patients examined, 31 were diagnosed with gingivitis, 38 with glossitis, 28 with leucoplakia, 37 with adult periodontitis, 25 with juvenile periodontitis, 36 stomatitis prothetica and 40 with stomatitis atrophica. In 196 patients (83.4 +/- 2.4%), fungi belonging to Candida species were detected. In the evaluation of Candida albicans strains (146) properties, bioMerieux API ZYM tests containing substrates for the detection of 19 hydrolases were used. All the investigated strains were characterized by the activity of 14 enzymes, i.e. phosphatase alcaline, esterase (C4), esterase lipase (C8), leucine and valine arylamidase, phosphatase acid, naphthol-AS-BI-phosphohydrolase, alpha galactosidase, beta galactosidase, alpha glucosidase, beta glucosidase, N-acetyl beta-glucosaminidase, alpha mannosidase and alpha fucosidase. Strains isolated from the oral cavity of patients with disease of periodontium and mucous membrane are characterised by the highest phosphatase acid activity. The greatest enzymatic activity is characteristic of Candida albicans isolated from patients with stomatitis atrophica or stomatitis prothetica, and the lowest in strains from gingivitis or juvenile periodontitis cases. Differences in the activity of hydrolases are statistically significant (p < 0.01) for: esterase (C4), leucine and valine arylamidase, phosphatase acid, naphthol-AS-BI-phosphohydrolase, beta glucosidase, N-acetyl-beta-glucosaminidase, of fungi isolated from patients with particular clinical diagnoses. PMID- 9750342 TI - Enzymatic oxidations in the biosynthesis of complex alkaloids. AB - The biosynthesis of complex alkaloids in plants involves enzymes that, due to high substrate specificity, appear to have evolved solely for a role in secondary metabolism. At least one class of these enzymes, the oxidoreductases, catalyze transformations that are in some cases difficult to chemically mimick with an equivalent stereo- or regiospecificity and yield. Oxidoreductases are frequently catalyzing reactions that result in the formation of parent ring systems, thereby determining the class of alkaloid that a plant will produce. The oxidoreductases of alkaloid formation are a potential target for the biotechnological exploitation of medicinal plants in that they could be used for biomimetic syntheses of alkaloids. Analyzing the molecular genetics of alkaloid biosynthetic oxidations is requisite to eventual commercial application of these enzymes. To this end, a wealth of knowledge has been gained on the biochemistry of select monoterpenoid indole and isoquinoline biosynthetic pathways, and in recent years this has been complemented by molecular genetic analyses. As the nucleotide sequences of the oxidases of alkaloid synthesis become known, consensus sequences specific to select classes of enzymes can be identified. These consensus sequences will potentially facilitate the direct cloning of alkaloid biosynthetic genes without the need to purify the native enzyme for partial amino acid sequence determination or for antibody production prior to cDNA isolation. The current state of our knowledge of the biochemistry and molecular genetics of oxidases involved in alkaloid biosynthesis is reviewed herein. PMID- 9750343 TI - The Arabidopsis ERECTA gene is expressed in the shoot apical meristem and organ primordia. AB - In Arabidopsis thaliana (L.) Heynh, the mutation in ERECTA is known to confer a compact inflorescence by a reduction in the lengths of internodes and pedicels. We analyzed the expression pattern of this gene during plant development. In situ hybridization and histochemical analysis using transgenic plants carrying chimeric gene fusions, with the ERECTA promoter fused to the beta-glucuronidase (GUS) gene, showed that ERECTA was predominantly expressed in the shoot apical meristems and organ primordia. ERECTA expression in the shoot apical meristem was weak early in plant development but increased with the transition from the vegetative to the reproductive growth phase. ERECTA was also strongly expressed in organ primordia and immature organs but weakly in mature organs. Thus, ERECTA was expressed in a cell-specific and developmentally regulated manner. In order to identify the regulatory mechanism responsible for the expression pattern of ERECTA, the cis-acting regions in the ERECTA promoter were defined by study of the expression of the chimeric genes that consist of the 5'- or internal deleted promoter and a GUS reporter gene in transgenic plants. The results showed that the essential cis-regulatory elements governing the spatially and temporally specific expression of ERECTA are located between positions -462 and -228 bp and between positions -228 and -153 bp with respect to the transcriptional initiation site. PMID- 9750344 TI - Gibberellin-photoaffinity labelling of two polypeptides in plant plasma membranes. AB - Two polypeptides of M(r) 68 kDa and 18 kDa were gibberellin (GA)-photoaffinity labelled in vitro in plasma membrane preparations from oat (Avena sativa L.) aleurone and from leaves and stems of wild-type and GA-sensitivity mutants of different species. Labelling of these polypeptides could be competed by biologically active, but not by inactive, GAs, indicating the likely biological significance of these interactions. On 2-dimensional gels the radiolabelled polypeptides were each resolved as one intensely labelled low abundance spot with a slightly lower pl form adjacent to it. There was a strong pH dependency for both labelling events, which correlated well with pH values at which GA are known to be most biologically active. A semi-dwarf GA-sensitivity mutant of sweet pea (Lathyrus odoratus L.), lb, showed reduced photoaffinity labelling of both polypeptides compared with the wild type, Lb. In the GA-insensitive Arabidopsis thaliana mutant gai, the level of labelling was the same as in wild type, GAI. This is the first report of GA-binding proteins in plant plasma membranes. Some preliminary sequence data are given for one of the labelled polypeptides. We discuss these mutants and consider their possible roles in GA perception or action. PMID- 9750345 TI - A cytokinesis-defective mutant of Arabidopsis (cyt1) characterized by embryonic lethality, incomplete cell walls, and excessive callose accumulation. AB - The genetic control of cell division in eukaryotes has been addressed in part through the analysis of cytokinesis-defective mutants. Two allelic mutants of Arabidopsis (cyt1-1 and cyt1-2) altered in cytokinesis and cell-wall architecture during embryogenesis are described in this report. Mutant embryos appear slightly abnormal at the heart stage and then expand to form a somewhat disorganized mass of enlarged cells with occasional incomplete walls. In contrast to the keule and knolle mutants of Arabidopsis and the cyd mutant of pea, which also exhibit defects in cytokinesis during embryogenesis, cyt1 embryos cannot be rescued in culture, are desiccation-intolerant at maturity, and produce cell walls with excessive callose as revealed through staining with the aniline blue fluorochrome, Sirofluor. Some cyt1 defects can be partially phenocopied by treatment with the herbicide dichlobenil, which is thought to interfere with cellulose biosynthesis. The distribution of unesterified pectins in cyt1 cell walls is also disrupted as revealed through immunocytochemical localization of JIM 5 antibodies. These features indicate that CYT1 plays an essential and unique role in plant growth and development and the establishment of normal cell-wall architecture. PMID- 9750347 TI - A starch-accumulating mutant of Arabidopsis thaliana deficient in a chloroplastic starch-hydrolysing enzyme. AB - The aim of this work was to identify enzymes that participate in the degradation of transitory starch in Arabidopsis. A mutant line was isolated by screening leaves at the end of the night for the presence of starch. The mutant had a higher starch content than the wild-type throughout the diurnal cycle. This accumulation was due to a reduction in starch breakdown, leading to an imbalance between the rates of synthesis and degradation. No reduction in the activity of endo-amylase (alpha-amylase), beta-amylase, starch phosphorylase, maltase, pullulanase or D-enzyme could be detected in crude extracts of leaves of the mutant. However, native PAGE in gels containing amylopectin revealed that a starch-hydrolysing activity, putatively identified as an endo-amylase and present in wild-type chloroplasts, was absent or appreciably reduced in the mutant. This is the first time that a specific enzyme required for starch degradation has been identified in leaves. PMID- 9750346 TI - Cloning and characterization of MS5 from Arabidopsis: a gene critical in male meiosis. AB - In this paper, we describe the cloning of the MS5 gene, a gene essential for male fertility in Arabidopsis. We previously defined the MS5 locus by characterizing an EMS-induced allele, ms5-1. We identified a new allele of MS5 (ms5-2) that was T-DNA-generated and used the T-DNA tag to clone the gene. Sequencing of mutant and wild-type alleles together with complementation of the ms5-1 mutant phenotype with a wild-type genomic clone confirmed the identity of the gene. Differences between the phenotypes of the two mutant alleles could be attributed to differences in mutant gene structure. The semi-dominant and dominant negative phenotypes of the ms5-2 mutant probably result from production of a truncated polypeptide. An unknown locus in Landsberg erecta can counteract the dominant negative phenotype of ms5-2. Mutations in MS5 cause the formation 'polyads'- tetrads with more than four pools of chromosomes after male meiosis. Similarities between the MS5 sequence and that of a number of proteins were found; two that may be significant were with a synaptonemal complex protein and with a regulatory subunit of a cyclin-dependent kinase. The MS5 gene is a member of a small gene family highly conserved amongst plant species. PMID- 9750349 TI - KAS IV: a 3-ketoacyl-ACP synthase from Cuphea sp. is a medium chain specific condensing enzyme. AB - cDNA clones encoding a novel 3-ketoacyl-ACP synthase (KAS) have been isolated from Cuphea. The amino acid sequence of this enzyme is different from the previously characterized classes of KASs, designated KAS I and III, and similar to those designated as KAS II. To define the acyl chain specificity of this enzyme, we generated transgenic Brassica plants over-expressing the cDNA encoded protein in a seed specific manner. Expression of this enzyme in transgenic Brassica seeds which normally do not produce medium chain fatty acids does not result in any detectable modification of the fatty acid profile. However, co expression of the Cuphea KAS with medium chain specific thioesterases, capable of production of either 12:0 or 8:0/10:0 fatty acids in seed oil, strongly enhances the levels of these medium chain fatty acids as compared with seed oil of plants expressing the thioesterases alone. By contrast, co-expression of the Cuphea KAS along with an 18:0/18.1-ACP thioesterase does not result in any detectable modification of the fatty acids. These data indicate that the Cuphea KAS reported here has a different acyl-chain specificity to the previously characterized KAS I, II and III. Therefore, we designate this enzyme KAS IV, a medium chain specific condensing enzyme. PMID- 9750350 TI - Over-expression of a tobacco homeobox gene, NTH15, decreases the expression of a gibberellin biosynthetic gene encoding GA 20-oxidase. AB - Ectopic expression of the homeobox gene, NTH15 (Nicotiana tabacum homeobox 15) in transgenic tobacco leads to abnormal leaf and flower morphology, accompanied by a decrease in the content of the active gibberellin, GA1. Quantitative analysis of intermediates in the GA biosynthetic pathway revealed that the step from GA19 to GA20 was blocked in transgenic tobacco plants overexpressing NTH15. To investigate the relationship between the expression of NTH15 and genes involved in GA biosynthesis, we isolated three cDNA clones from tobacco encoding two types of GA 20-oxidase and a 3 beta-hydroxylase. RNA gel blot analysis revealed that the expression of one gene (Ntc12, encoding GA 20-oxidase), which in wild-type tobacco plants was abundantly expressed in leaves, was strongly suppressed in the transformants. The expression level of Ntc12 decreased with increasing severity of phenotype of transgenic tobacco leaves. The abnormal leaf morphology was largely overcome by treatment with GA20 or GA1 but not by GA19. These data strongly suggest that overexpression of NTH15 inhibits the expression of Ntc12, resulting in reduced levels of active GA and abnormal leaf morphology in transgenic tobacco plants. In situ hybridization in wild-type tobacco revealed that expression of Ntc12 occurred mainly in the rib meristem, cells surrounding the procambium and in leaf primordia. Expression was not seen in the tunica, corpus and procambium, tissues in which NTH15 was predominantly expressed. The contrasting expression patterns of these genes may reflect their antagonistic functions in the formation of lateral organs from the shoot apical meristem. PMID- 9750353 TI - Gain of function assays identify non-rol genes from Agrobacterium rhizogenes TL DNA that alter plant morphogenesis or hormone sensitivity. AB - This study tested the morphogenetic potential of 15 open reading frames of the TL DNA of Agrobacterium rhizogenes strain HRI. These open reading frames were expressed individually under the control of the 35S RNA promoter in transgenic tobacco plants (Nicotiana tabacum L.). Expression of three T-DNA loci, ORF3n, ORF8 and ORF13, alters plant morphogenesis or the response of transgenic tissues to plant hormones. ORF3n transgenic plants are characterized by retarded flowering, altered internode elongation, altered leaf shape and, in particular, leaf tip necrosis. ORF3n and ORF8 expression reduces the sensitivity to auxin and cytokinin in combination or auxin alone. Tetracycline-dependent expression of ORF13 overcomes a selection of low levels of expression during plant regeneration and reveals a strong inhibitory effect of the ORF13 gene product on cell division and cell elongation. We conclude that the A. rhizogenes TL-DNA harbors genetic information that is important for pathogenicity apart from the well studied rol genes. We propose that these genes play mainly a negative regulatory role during pathogenesis. Moreover, these loci might be relevant to successful infections in specific host plants. PMID- 9750352 TI - Two dehydration-inducible transcripts from the resurrection plant Craterostigma plantagineum encode interacting homeodomain-leucine zipper proteins. AB - The molecular dissection of desiccation tolerance in the resurrection plant Craterostigma plantagineum led to the isolation of two dehydration-stress inducible homeo-domain-leucine zipper genes (CPHB-1 and -2). When the coding region of CPHB-1 was used as bait in the yeast two-hybrid system, the ability of CPHB-1 to form homodimers was demonstrated. The two-hybrid system was also used to isolate CPHB-2, which heterodimerises with CPHB-1. Both transcripts are inducible by dehydration in leaves and roots, but steady state levels vary in response to exogenously applied ABA. Although expression of CPHB-1 is not inducible by ABA, the transcript level of CPHB-2 increases during ABA-treatment. Both genes are expressed at very early stages of dehydration and thus may be involved in the regulation of gene expression during dehydration. CPHB-1 and -2 differential expression in response to ABA suggests that they act in different branches of the dehydration-induced signalling network. In vitro binding studies revealed that CPHB-1 specifically binds to the pseudopalindromic sequence CAAT(C/G)ATTG. Using this element for in vitro binding studies with nuclear proteins from dehydrated leaves, an inducible DNA-protein complex was identified. PMID- 9750354 TI - The untranslated leader sequence of the barley lipoxygenase 1 (Lox1) gene confers embryo-specific expression. AB - The barley lipoxygenase 1 (Lox1) gene encodes a protein expressed in embryos during grain development and germination and in leaves after methyl-jasmonate (MeJA) treatment. Transient gene expression assays in germinating barley embryos were used to identify cis-regulatory elements involved in the embryo-specific expression of the Lox1 gene. Analysis of transcriptional or translational fusions between Lox1 5' upstream sequences and the gusA reporter gene indicated that the 5'-untranslated leader sequence was involved in embryo-specific expression. Replacement of the leader sequence from the aleurone-specific Chi26 gene with the Lox1 leader sequence resulted in a chimeric gene expressed at high levels in embryo as well as in aleurone cells. Insertion of the Lox1 leader sequence between the 35S minimum promoter (A domain -90/+8) and the gusA reporter gene greatly enhanced promoter activity in a tissue-specific manner. Deletion/replacement analysis of the Lox1 leader sequence, combined with transient expression in germinating embryos and in vitro transcription/translation assays, suggests that the Lox1 leader sequence contains cis-elements regulating qualitative (tissue-specific) and quantitative gene expression. PMID- 9750355 TI - Stacks on tracks: the plant Golgi apparatus traffics on an actin/ER network. AB - We have visualized the relationship between the endoplasmic reticulum (ER) and Golgi in leaf cells of Nicotiana clevelandii by expression of two Golgi proteins fused to green fluorescent protein (GFP). A fusion of the transmembrane domain (signal anchor sequence) of a rat sialyl transferase to GFP was targeted to the Golgi stacks. A second construct that expressed the Arabidopsis H/KDEL receptor homologue aERD2, fused to GFP, was targeted to both the Golgi apparatus and ER, allowing the relationship between these two organelles to be studied in living cells for the first time. The Golgi stacks were shown to move rapidly and extensively along the polygonal cortical ER network of leaf epidermal cells, without departing from the ER tubules. Co-localization of F-actin in the GFP expressing cells revealed an underlying actin cytoskeleton that matched precisely the architecture of the ER network, while treatment of cells with the inhibitors cytochalasin D and N-ethylmaleimide revealed the dependency of Golgi movement on actin cables. These observations suggest that the leaf Golgi complex functions as a motile system of actin-directed stacks whose function is to pick up products from a relatively stationary ER system. Also, we demonstrate for the first time in vivo brefeldin A-induced retrograde transport of Golgi membrane protein to the ER. PMID- 9750356 TI - Why do microbes have toxins? PMID- 9750357 TI - Bacterial phospholipases and intracellular growth: the two distinct phospholipases C of Listeria monocytogenes. PMID- 9750358 TI - Pore-forming bacterial cytolysins. PMID- 9750359 TI - Cholera toxin and related enterotoxins as potent immune modulators. PMID- 9750360 TI - Cyanobacterial toxins and human health. PMID- 9750361 TI - Marine biotoxins. PMID- 9750362 TI - A risk assessment approach for food-borne Bacillus cereus and its toxins. PMID- 9750363 TI - Recent studies of mycotoxins. PMID- 9750364 TI - Toxigenic Escherichia coli. PMID- 9750365 TI - Botulinum neurotoxins: mode of action and detection. PMID- 9750366 TI - A review of analytical methods for the detection of bacterial toxins. PMID- 9750367 TI - Kits for the detection of some bacterial food poisoning toxins: problems, pitfalls and benefits. PMID- 9750368 TI - The role of streptococcal toxins in disease. PMID- 9750369 TI - Staphyloccal alpha toxin. PMID- 9750370 TI - Bacterial phospholipases. AB - The phospholipases are a diverse group of enzymes, produced by a variety of Gram positive and Gram-negative bacteria. The roles of these enzymes in the pathogenesis of infectious disease is equally diverse. It is only recently that molecular genetic approaches have allowed data to be obtained which indicates the role of these enzymes in the disease process. In the case of some pathogens phospholipases play an overriding role in disease. Roles for these enzymes have been demonstrated in the pathogenesis of disease caused by extracellular and intracellular pathogens and by disease caused by pathogens which enter via the respiratory tract, the intestinal tract or after traumatic injury. Some of the mechanisms by which phospholipases C affect tissues in vitro or ex vivo are understood but, in the main, the mechanisms by which phospholipases C affect tissues in vivo are not known. A key event, which can determine the extent of involvement of phospholipases in the disease process, is the interaction of the enzyme with phospholipids in eukaryotic cell membranes. Whilst progress has been made in understanding the molecular basis of these interactions, the process is far from understood. Two theories attempt to explain the reasons why only some phospholipases C are membrane active. In general, the membrane active enzymes are able to hydrolyse both phosphatidylcholine and sphingomyelin and appear to have mechanisms which allow them to interact with membrane phospholipids. The structural differences between phosphatidylcholine and sphingomyelin lie within the fatty acyl chain/ester bond region which would be partially embedded in the membrane bilayer. Therefore, there may be a common explanation for membrane interaction and recognition of both phospholipid types. The value of this information will be several fold. The demonstration of the role of these enzymes in disease will allow the development of vaccines or therapeutics which block the effects of these enzymes. In this context it is worth bearing in mind that eukaryotic phospholipases C, which play key roles in many inflammatory and autoimmune diseases, are the subject of intense study by the pharmaceutical industry. Some of the bacterial toxins are potent cytotoxic agents and this has encouraged some workers to explore the possibility that immunotoxins can be developed (Chovnick et al. 1991). Purified recombinant phospholipases C will continue to be used in the study of cell membranes, and the increasing numbers of enzymes with different substrate specificities will enhance their application. PMID- 9750371 TI - Bacterial toxins which perturb ciliary function and respiratory epithelium. PMID- 9750372 TI - [The EUROPOP Project. European Programme of Occupational Risks and Pregnancy Outcomes]. AB - Efforts to obtain an objective view of the working and living conditions of European women and in particular the influence of these conditions on the course of pregnancy were the reason why in 1994 within research activities of the EC a project EUROPOP (European Programme of Occupational Risk and Pregnancy Outcome) was adopted and started. Seventeen countries incl. the Czech Republic were asked to participate. The research proper was conducted in 57 maternity institutions. In the Czech Republic the Olomouc region was selected with the Gynaecological and Obstetric Clinic in Olomouc as the coordinating centre. All 13 gynaecological and obstetric departments of the Olomouc catchment area were included in the trial. PMID- 9750374 TI - [Levels of lymphocyte subpopulations in peripheral fetal blood in Rh(D) erythrocyte isoimmunization in pregnancy]. AB - INTRODUCTION: In the foetus in utero predominates a considerable percentage of immunologically so-called naive lymphocytes of the T and B series. The objective of the presented work was to assess by means of flow cytometry and labelled monoclonal antibodies quantitative changes in cell sub-populations of the foetal immune system before the foetus is altered by severe heamolysis as a result of Rh(D) isoimmunization. METHOD: The authors obtained by intrauterine puncture of the umbilical cord peripheral blood from 10 foetuses with isoimmunization during the 23rd-35th week of gestation, confirmed by the direct Coombs test and with a mean haematocrit value of 30.8% +/- 8.02. These findings were compared with values in the peripheral blood stream in a control group of 35 foetuses during the 18th-39th week of pregnancy with normal intrauterine development (haematocrit 34.2% +/- 5.87). The authors assessed the haemogram and CD signs in the lymphocyte population. RESULTS: In the peripheral bloodstream of foetuses with erythrocyte isoimmunization the authors did not detect, as compared with the control group (p > 0.01), a lower haemoglobin level (10.6 +/- 2.77 g/dl vs. 11.9 +/- 2.03 g/dl) a lower haematocrit (30.8% +/- 8.02 vs. 34.2% +/- 5.87) and fewer leucocytes 5.1 +/- 2.39 x 10(9)/l versus 6.97 +/- 3.29 x 10(9)/l. In foetuses with Rh(D) isoimmunization the authors found a higher percentage of T(CD3+) lymphocytes (79.0% +/- 11.23 vs. 73.7% +/- 12.79, but did not prove an increase of activated T lymphocytes (%DR+ from CD3+) (1.0% +/- 0.52 vs. 1.3 +/- 0.58). The percentage ratio of T helper cells (CD+) was higher than in the control group (61.0% +/- 10.25 vs. 56.1% +/- 12.45). In foetuses with Rh(D) isoimmunization there was no difference in the ratio of CD8 positive cells (24.1% +/- 8.23 vs. 23.6% +/- 7.26). Suppressor T cells (CD8+CD11b+) were fewer (4.1% +/- 1.24 vs. 7.5 +/- 11.23) than in the control group. The number of NK cells in foetuses with Rh(D) isoimmunization was 5.1% +/- 3.26 vs. 6.9% +/- 3.86, in isoimmunized foetuses there is a higher ratio of so-called naive T helper cells Th1 (CD4+ CD45 RA+) 49.3% +/- 12.71 vs. 43.0% +/- 12.88. When assessing naive B1 cells (CD19+CD5+), the authors did not find a difference between the two groups (10.4% +/- 6.20 vs. 9.06% +/- 12.1). The ratio of CD4: CD8 in the group with isoimmunization was higher than in the control group (3.1 +/- 1.39 vs. 2.5 +/- 1.13). CONCLUSION: In isoimmunized foetuses in the initial stages of haemolysis no detactable immune response with significant changes in the lymphocyte sub populations was found. PMID- 9750373 TI - [Diagnosis and therapy of erythrocyte alloimmunization in pregnancy]. AB - INTRODUCTION: By preventive administration of anti-D globulin the number of cases of Rh isoimmunization declines steadily. Severe untreated isoimmunization may lead via foetal hydrops to intrauterine death, sometimes already during the 18th 19th week of gestation. The purpose of prenatal diagnosis in pregnant women with isoimmunization is to assess the danger or affection of the foetus, its prognosis and the mode of monitoring of the foetus. It is necessary to decide in time on intrauterine therapy by transfusion of erythrocyte mass and to assess the optimal time of delivery with regard to the risk of prematurity and foetal erythroblastosis, as well as with regard to intrauterine therapy. The objective of the present work was to test the protocol in the treatment of erythrocytic isoimmunization of the foetus. METHOD: During the period between January 1991 and October 1997 the authors investigated two groups of pregnant women: with a hydropic (n = 5) and non-hydropic (n = 20) foetus at the onset of treatment. In both groups amniocentesis and umbilical puncture were indicated. The authors investigated the number of cordocenteses and the volume of transfused blood per pregnancy, the number of complications and their type, gestation age of the foetuses on delivery, their birth weight, the condition of the neonates after delivery and on discharge to home care. RESULTS: During the mentioned period the authors administered 70 intraumbilical transfusions to 25 foetuses. The transfusion was not repeated more than eight times. The baseline haematocrit of non-hydropic foetuses was 26 (14-34), treatment was started on average during the 28th week (23rd-33rd). Pregnancy in women with a non-hydropic foetus was terminated during the 35th (27th-40th) week, with a mean weight of the foetuses of 2439 g (870-3520). Of 25 treated foetuses 6 were hydropic (24%) at the onset of treatment. The initial haematocrit of hydropic foetuses was 10.7 (4-19.8), treatment was started on average during the 28th (23rd-33rd) week. Pregnancy of women with hydropic foetuses was terminated during the 30th (25th-36th) week, the mean birth weight being 1838 g (660-3500). DISCUSSION: The very favourable therapeutic results in non-hydropic foetuses are in great contrast with the therapeutic results of moribund hydropic foetuses. CONCLUSION: The basic prerequisite of successful treatment by intraumbilical transfusion is to concentrate risk pregnancies in specialized centres with a high standard neonatological team for intensive care of pathological neonates. PMID- 9750375 TI - [The effect of endometrial infection on embryo implantation in the IVF and ET program]. AB - The results of the IVF and ET programme depend on a number of known and hitherto unknown factors. One of them is implantation of the embryo. Only 10-15% of the embryos transferred into the uterus are implanted. The most important factors affecting implantation are quality of the embryo and receptivity of the endometrium. The receptivity of the endometrium depends on hormonal changes, vascularization of the uterus, infectious environment and other factors. Part of them can be influenced to a certain extent. Infection of the endometrium as the cause of unsuccessful implantation could be resolved by therapeutic administration of antibiotics. The objective of the present work was to prove the possible effect of bacterial contamination of the endometrium on results of IVF + ET programmes. During 1996-1997 120 sterile women included in the IVF + ET programme were examined at the First Gynaecological and Obstetric Clinic (transport system-Pronatal, Prague). In all women the author performed during the cycle preceding the stimulated one (the IVF cycle proper) microabrasion of the endometrium by a method which prevents as much as possible contamination of the specimen by the endocervical flora. A smear from the endocervix and vagina was made. The samples were examined for the presence of aerobic and anaerobic flora, Chlamydiae. The endometrium was evaluated also from the histopathological aspect. Results of the examination of the endometrium: 62% of the specimens were positive on cultivation. Most frequently Pseudomonas spec. and Staph. epidermidis were found. Of 34 women who became pregnant 10 had a positive cultivation (29.4%). Of 86 women where pregnancy was not achieved, cultivation was positive in 64 (74.4%). This difference is statistically significant. The same cultivation finding from the endocervix and endometrium was observed only in 21% patients. From the trial where more than twice as many positive cultivations were recorded in non-conceptive cycles as compared with conceptive ones, the following preliminary conclusions can be drawn: 1. Elimination of endometrial infection has its place in the preparation of patients in the IVF + ET programme. 2. It is advisable to treat with antibiotics a positive cultivation of the endometrium or administer antibiotics prophylactically (62% positive) to all patients during a stimulated IVF cycle. PMID- 9750376 TI - [Changes in the position of the ureterovesical junction during maximal voluntary contractions and during maximal vaginal electric stimulation of the pelvic floor muscles]. AB - The objective of the study was to evaluate and compare the effect of the maximal voluntary muscle contraction of the pelvic floor (PFM) and contractions of the PFM evoked by maximal electric stimulation using an electrostimulation apparatus Conmax by monitoring the position of the urethrovesical junction by ultrasound. The trial comprised 20 women with confirmed stress incontinence of urine. With the patients in a supine position with abducted lower extremities an electrostimulation probe was inserted into the vagina. This was followed by perineal ultrasound (US) examination using an ACUSON 128 XP-10 apparatus and a convex tube 5 MHz. The ultrasound examination was made using the electrostimulation probe--at rest and during maximal voluntary contraction of the PFM. This was followed by maximal electric stimulation and after five minutes during stimulation the US examination was repeated. It was performed also during maximal electric stimulation (MES) concurrently with maximal voluntary contraction of the PFM. For electrostimulation a Conmax appartus was used. The applied frequency was 50 Hz, amplitude from 0 to 90 mA (grade 0-6), duration of pulse 0.75 ms. The maximum intensity of stimulation was determined by the patient, i.e. when stimulation was not yet painful. During US the authors investigated the gamma angle, i.e. the angle between the axis of the symphysis and the connecting line between the UV junction and the lower borderline of the symphysis. The mean difference of the gamma angle during voluntary contraction of the PFM and at rest was 13.6. During contraction caused by maximal electric stimulation of the PFM and at rest this difference was 21.3. The difference did not differ significantly during maximal electric stimulation of the PFM and during maximal electric stimulation and voluntary contraction of the PFM. From the trial ensues that contraction of the pelvic floor muscles during maximal electric stimulation is stronger as compared with the intensity of contraction caused by maximal voluntary contraction. The results confirm the favourable therapeutic effect of MES muscles of the pelvic floor in the treatment of urinary incontinence in women. These changes help to increase the muscular tonus and contractibility of pelvic floor muscles and thus promote also elevation of the neck of the urinary bladder. Elevation of the neck of the urinary bladder promotes normalization of intraabdominal transfer of pressure to the proximal urethra. PMID- 9750377 TI - [Diagnosis of partial hydatidiform mole using molecular genetics]. AB - The aim of our study was to confirm the diagnosis of partial hydatidiform mole (PMH) and to determine mechanism of PMH development by chromosomal DNA analysis. In our study we analysed 8 cases of morphologically and histologically confirmed PMH. Their karyotype was determined by using methods of direct, 24-hour and a long-term tissue culture. The restriction fragment length polymorphisms (RFLP) method was used for DNA analysis of PMH chorionic villi and from progenitor lymphocytes in peripheral blood. All cases were triploid. DNA analysis revealed in six cases one maternal and two paternal RFLP bands, and in two cases two maternal and one paternal RFLP bands. PMID- 9750378 TI - [Urologic complications of gynecologic surgery and their treatment in our clinical data]. AB - In 19 women (aged 27-66 years, mean 44.6 years) a retrospective analysis of the urological complications suffered during gynecological surgery was performed. 14 (73.6%) patients had ureteral injuries with concomitant bladder lesions in two. In the remaining 4 (21%) bladder injury only was present including a woman with a simultaneous urethral lesion. In the postoperative period 3 ureteric, 5 vesicovaginal, and one vesicouterine fistula developed. Intraoperative diagnosis followed by immediate surgical repair of the ureteric and bladder lesions was performed in 2 (10.5%) patients only. Except one case ureteral injury was unilateral, and predominantly right-sided (10:5). One patient suffered injury of one of the ureters in a complete ureteral duplication. The most frequent side of ureteral injury was at the level of the uterine artery with complete ligature of the ureter in 10 (71.4%), and partial in 4 (28.6%) patients. Percutaneous nephrostomy as a temporary diversion was performed in 8 (57.1%) patients. Definitive treatment involved simple Paquin ureteral reimplantation in 5 patients, Boari's procedure in 3, and psoas hitch technique once. Ureteral lysis only was sufficient in 3 patients. The surgical treatment including repair of vesical fistulas was in all cases successful. PMID- 9750379 TI - [Treatment of male infertility after oncologic therapy: preventive autologous sperm preservation versus testicular biopsy and oocyte fertilization with testicular spermatozoa]. AB - The prognosis of oncological therapy of some malignancies is favourable. Unfortunately the majority of young patients remain sterile. Sperm cryopreserved before treatment was successfully used in IVF-ICSI (in vitro fertilization intracytoplasmic sperm injection) cycles. Testicular biopsy (TESE) was performed after successful oncological treatment in three men without previous sperm cryopreservation. No spermatozoa were found for oocyte fertilization. TESE could not ensure sperm recovery in all patients after oncological treatment. Sperm cryopreservation should be highly recommended to all patients before any treatment is offered. PMID- 9750380 TI - [Ultrasonography of pelvic floor muscles in women with urinary stress incontinence]. AB - Ultrasound examinations have become since beginning of the eighties one of the auxiliary examination methods in urogynaecology. Evaluation of the position and mobility of the neck of the urinary bladder practically replaced lateral chain urethrocystography. With the improving differentiating capacity of ultrasound equipment it is possible to visualize some periurethral and paravaginal structures which participate in the support of the urethra and urethrovesical junction by paravaginal and periurethral structures in women (fig. 1). In recent years many papers were published where for visualization nuclear magnetic resonance (NMR) is used [1, 12, 19]. The functional and physiological condition of these tissues is assessed by physical, urodynamic and ultrasonographic examinations and also by cystourethroscopy and manometry [7, 11, 10, 13, 14, 15, 16]. Despite this the greater part of anatomical knowledge of the supporting apparatus is derived from pathological studies and peroperative observations. PMID- 9750382 TI - [Occurrence of congenital hydrocephalus in the Czech Republic 1961-1995]. AB - During the period between 1961-1965 the authors investigated the incidence of congenital hydrocephalus (CH) on the territory of the Czech Republic (CR). From a consecutive series of 5,137,907 births in the CR during 1961-1995 2,288 cases of CH were diagnosed incl. 2,024 cases in born infants and 164 were diagnosed prenatally and the pregnancy was terminated. The mean incidence of CH in born children was during the investigation period 3.99 per 10,000 liveborn infants, after adding cases with a prenatal diagnosis the mean incidence was 4.39 per 10,000 liveborn infants. The success rate of prenatal diagnosis of congenital hydrocephalus was on average almost 40% detected cases in the CR during 1989 1995. PMID- 9750381 TI - [Gestational diabetes--a model trial. The effect of alloxan diabetes on the spectrum of fatty acids in the blood, liver and brain in laboratory rats]. AB - In alloxan-induced diabetic rats (Wistar, females, age 3-4 months of postnatal life) the large spectrum of fatty acids in blood serum, brain cortex, medulla oblongata and liver was studied. The fatty acids, using gas chromatography, were detected as methyl esters and the methods were published previously (Smidova and al. 1994). Alloxan (Merck) administered i.p., 140 mg/kg body weight, caused immediately elevation (three times) of blood sugar levels. On the 13th day the rats were killed. The results are as follows: a) In blood serum alloxan diabetes of cca two weeks duration caused a significantly increased participation of saturated FA and decreased participation of both polyunsaturated FA (n-3 and n 6). b) In brain cortex no differences between controls and diabetic rats in the indicated groups of FA were found. c) In the medulla oblongata an increased participation of polyunsaturated fatty acids n-6 was established. d) In hepatic tissue the increased participation of saturated FA as well as a decreased participation of FA n-6 was described. Analysing the main groups of FA we found especially in n-3 and n-6 FA several significant changes in single FA (a smaller pool of arachidonic acid in blood serum as well as in liver, decreased participation of docosahexaenoic acid in the brain cortex, etc.). The purpose and possible consequences of such changes are discussed. PMID- 9750383 TI - [Squamous cell carcinoma markers in neoplasms of the uterine cervix]. PMID- 9750384 TI - [Cryopreservation of embryos after intracytoplasmic sperm injection (ICSI)]. PMID- 9750385 TI - [Chlamydia trachomatis--the epidemiologic situation]. PMID- 9750386 TI - [Short-term administration of Danol before endometrial ablation]. PMID- 9750387 TI - [The premature menopause syndrome (premature ovarian failure)]. PMID- 9750388 TI - [Diagnostic approach in postmenopausal hemorrhage]. PMID- 9750389 TI - [Transvaginal echocardiography--successful imaging and quantitative analysis of fetal cardiac structures in the 13th to 17th week of pregnancy]. PMID- 9750390 TI - [Meropenem in the treatment of severe gynecologic infections]. PMID- 9750391 TI - [Local administration of antibiotics (garamycin implant/Schwamm) in surgical treatment of suppurative complications in gynecology]. PMID- 9750392 TI - [Comments on the nation-wide research project on contraceptive behavior in the population of the Czech Republic]. PMID- 9750393 TI - [Placental abruption with development of disseminated intravascular coagulation]. PMID- 9750394 TI - [Postmenopausal osteoporosis]. PMID- 9750395 TI - [Receptors and mechanisms of action of estrogen in bones]. PMID- 9750396 TI - [Genetic risks associated with treatment of marital infertility using intracytoplasmic sperm injection]. PMID- 9750397 TI - [Systemic lupus erythematosus and the sex hormones]. PMID- 9750398 TI - [Cost-benefit in perinatology--I. General views of western economists]. AB - The mutual cost-benefit relationship of any activity is the main principle of the market mechanism where both these variables are expressed in monetary terms. According to western economists this system cannot be applied, or if so only partly, in the sphere of health services because health benefit can be only in exceptional instances expressed in monetary terms. This is why they recommended for evaluation of health care, incl. perinatal care, other economic models where the health benefit is evaluated by saved lives or improvement of the health status. With this aspect in mind the author described five recommended models for the evaluation of alternative procedures and for evaluation of programmes in perinatology. Each of them is supplemented by examples from clinical practice. Most frequently different views are held on the effectivity of costly screening programmes for early detection of threatened foetuses and intensive care of these neonates, i.e. procedures which are the main cause of rising costs of perinatal care. The author discusses reasons for controversial views on contemporary economic problems in perinatology. PMID- 9750399 TI - [Cost-benefit in perinatology--II. Its application to conditions in the Czech Republic]. AB - Using western economic models evaluating cost/benefit in perinatal care, the author analyzed on examples of undesirable perinatal outcomes the health benefit as a result of extension of screening and therapeutic procedures in the Czech Republic in 1990-1996 as well as the efficacy, effectivity and efficiency of these procedures. The results of analyses provided evidence that extension of the above procedures led to a decline of undesirable outcomes, different for each of the five investigated types, but at the same time a considerable increase of costs was recorded which exceed resources available at present for health services in the Czech Republic. Further development of perinatal care which will have to overcome this discrepancy without jeopardizing the present high standard of perinatal care will have to consider a compromise which will achieve a balance between present professional knowledge and possibilities, the moral aspect, demands of society and available funds. PMID- 9750400 TI - [Chlamydia trachomatis and its role in female infertility]. AB - Infertility is a worldwide problem. 15-20% married couples are childless though they want children. The tubal factor is the most frequent cause of female infertility. Chlamydia trachomatis is one of the most frequent pathogenic organisms which cause tubal occlusion. From January 1995 till June 1997 at the 1st Dept. Gynecol. and Obstetric, 1st Fac. of Medicine 327 infertile women were examined divided into three groups (sterile, included in the IVF programme on account of the tubal factor and those included in the programme for other reasons). In all an antigenic examination from the endocervix and serological examination of antichlamydia antibodies was made. In 73 women in the IVF programme by microabrasion of the endometrium material for detection of Chlamydia infection was obtained by PCR. During sampling from the uterine cervix the appearance of the portio uteri, discharge, haemorrhage and pain on examination were evaluated. For statistical evaluation the chi square test was used. In 61% patients anamnestic IGG antibodies were detected suggesting a past or present Chlamydia infection. In women included in the IVF programme on account of the tubal factor there was a significantly higher IgG positivity (92%). Of ten women who suffered from acute chlamydial cervicitis only in one the infection was detected during microabrasion of the endometrium. 65% antigen positive patients had complaints during the examination (discharge, bleeding, pain). The authors discuss the impact of the assembled results from the aspect of impaired fertility of women in the Czech Republic. PMID- 9750401 TI - [Treatment of chlamydial urogenital infections]. AB - Chlamydia trachomatis is the most frequent sexually transmitted bacterial pathogen in developed countries [3, 12, 13]. The position is similar in the Czech Republic. Depending on the group of examined women active Chlamydia infection varies between 10 and 23%. The increasing incidence of urogenital Chlamydia infections and improving diagnostic possibilities call for adequate treatment. Correct treatment of urogenital infections caused by Chlamydia trachomatis is very important for the prevention of undesirable sequelae of inflammations of the lesser pelvis, subsequent risk of GEU, sterility, prevention of premature delivery and possible infection of the neonate. When starting treatment, selecting a suitable antibiotic and deciding on the therapeutic strategy it is important to select an antibiotic with regard to its efficacy, the epidemiological situation, regional sensitivity of the infectious agent, toxicity and tolerance of the antibiotic, to its bacteriostatic or bactericide action, and last not least, also its price. Despite selection of a suitable antibiotic sometimes treatment fails. For treatment of urogenital chlamydial infections tetracyclin and macrolid antibiotics are recommended or quinolone chemotherapeutic agents of the third generation. Tetracyclines are broad spectrum antibiotics with bacteriostatic action. As to oral forms doxycycline, tetracycline and oxytetracycline are used. The most frequent undesirable effects during treatment are nausea, vomiting, diarrhoea and abdominal pain. Tetracycline antibiotics are contraindicated in children under 8 years, during pregnancy and lactation and in case of sensitivity to this group of drugs. Macrolids are antibiotics with a medium broad antibacterial spectrum with bacteriostatic action. Macrolids of the first generation have a low antibacterial activity. They have a short biological half-life, not always a good tolerance, and serious clinically important drug interactions may develop. The most frequently used preparations of the first generation include erythromycin, josamycina and spiramycin. Macrolids of the second generation, azitromycin, roxitromycin and claritromycin lack the above negative properties. The most frequent undesirable effects after administration of macrolids include nausea and vomiting. Considerable differences were found in particular between different preparations containing erythromycin. Macrolids of the second generation have only slight undesirable gastrointestinal effects. Macrolid antibiotics are contraindicated in case of sensitization to this group, in severe hepatic disorders and great care must be taken in the treatment of pregnant women. Quinolone chemotherapeutic agents of the third generation, ciprofloxacine, enoxacine, ofloxacine and pefloxacine are synthetic drugs with a broad antibacterial spectrum which act on systemic infections. On oral administration they are rapidly absorbed and the blood and tissue concentrations are sufficiently effective. In the treatment of urogenital Chlamydia infections they are useful in the treatment of chronic infections after failure of previous macrolid and tetracycline therapy. The most frequent undesirable side-effects include nausea, vomiting, meteorism, diarrhoea, tinnitus, headache, changes of mood, allergic skin reaction. They are contraindicated in hypersensitivity to quinolone chemotherapeutic preparations, in children and adolescents under 18 years, during pregnancy and lactation. The objective of the present study was to evaluate different therapeutic patterns, their efficacy and tolerance. PMID- 9750402 TI - [Immunoscintigraphy in the diagnosis of ovarian carcinoma]. AB - The authors define the place of immunoscintigraphy (IS), a diagnostic method which differs qualitatively from the usual examination methods (US ultrasonography, CT-computed tomography) in the management of ovarian cancer when monitoring the response to therapy and for detection of subclinical manifestations of the disease. The author explains the principle of the method which involves a highly specific antigen-antibody reaction. Possibilities to influence the accuracy of the examination and results of a clinical perspective study are submitted the objective of which was to test the possible use of this diagnostic method under our conditions. In a group of 66 women (with an established diagnosis of ovarian cancer or a newly detected ovarian tumour) 76 IS examinations were made and the finding was confirmed by biopsy. The results of the histological examination were also a criterion of the accuracy of IS and CT examinations resp. The material for biopsy was obtained on operation, second look operation, by fine needle biopsy or on necropsy. The authors evaluate 65 IS examinations. The accuracy of the examination was evaluated as very satisfactory (84%), the sensitivity 89%, specificity 79%, there was a minimal number of falsely positive findings (9.2%) and falsely negative ones (6.2%). An analysis of possible reasons of failure of IS examinations is made. The authors evaluate the contribution of IS and its place among diagnostic methods in the treatment and follow up of women with ovarian cancer. PMID- 9750403 TI - [Treatment of infertility in non-obstructive azoospermia using the TESE (testicular sperm extraction) method--clinical study]. AB - Testicular sperm extraction (TESE) was performed in 27 men in 30 cycles. All men were examined for genetics, serum hormonal status, biochemical status of semen samples. All men were examined by an urologist. No prognostic evaluation able to provide information about the prognosis of TESE procedure was found. Even a high FSH level, testicular hypotrophy or previous histological examination cannot exclude any patient from testicular biopsy. PMID- 9750404 TI - [Detection of cadmium and zinc in the blood and follicular fluid in women in the IVF and ET program]. AB - Scientific and technical development contributed on the one hand greatly to easier living conditions, on the other hand there is however substantially greater danger threatening not only the life of man but also of nature, of the entire living environment. At present the problem of the possible part played by this phenomenon in reproductive disorders is in the foreground. Via the food chain heavy metals, toxic trace elements and polyhalogenic hydrocarbons penetrate into the organism. They accumulate in the organism and thus also in the reproductive tract and may have an impact on fertility. Some elements with a possible negative impact on reproductive health are in such low concentrations in tissues that only contemporary methods of their detection make it possible to map their presence in the organism. They are called trace elements. Toxic ones comprise cadmium, mercury, lead and arsenic. The mechanism of the negative action of cadmium in the organism is most probably due to its competition with the vitally important trace element--zinc. It was therefore the objective of the present investigation to trace the presence of cadmium and zinc in the organism of 100 sterile women included in an IVF programme: in blood and follicular fluid (i.e. in a medium which surrounds the gamete--the oocyte) and to follow up their concentrations in relation to achievement pregnancy. Cadmium and zinc in blood and follicular fluid were assessed on a mass spectrometer with induction bound plasma as the source of ions (ICP-MS), Varian Co. produced in 1994. The assessed mean levels (microgram/l) of cadmium in blood (2.88 s 2.71) and follicular fluid (1.25 s 0.55) in the group of conception cycles did not differ significantly from mean blood levels (2.82 s 2.22) and follicular levels (1.16 s 0.55) of non conception cycles. The mean zinc levels in blood and follicular fluid did not differ either in the group of conception and non-conception cycles. Very significant are the differences in the blood and follicular fluid levels, the levels in follicular fluid being significantly lower. We may speak of a protective barrier of the oocyte formed by the follicle (probably the cells of the granulosa) against blood. Thus no relationship was found between the cadmium concentration in blood and follicular fluid of women where pregnancy was achieved and non-pregnant women. The possible cause of fertility disorders in conjunction with toxic elements is probably in damage of the granulosa cells and thus their dysfunction as regards production of steroid hormones with full impact on female fertility (hormone disruptors). PMID- 9750405 TI - [Effect of oocyte transportation (IVF transport) on effectiveness of the IVF and ET program]. AB - The programme of in vitro fertilization helps at present an ever increasing number of married couples to resolve successfully the problem of sterility. It is however a method pretentions technically, as regards time, material and in particular funds. This applies above all to embryological laboratories. The transport system of the IVF programme makes possible care of patients indicated for sterility treatment by the IVF method in departments without their own embryological laboratory and thus it is possible to include more patients in the programme and save money. In the author's department the first two stages of the IVF programme are implemented, i.e. stimulation of the ovaries and aspiration of follicular fluid. Then this material is taken in a transport box of the K-system Co., Denmark, type G-81E to a centre with full accreditation where the subsequent stages of the IVF programme take place (selection of the oocyte, fertilization, cultivation of embryos and embryo transfer). The transport system has operated now for three years (1995-1997) and at present the authors evaluate the system. The fertility rate and pregnancy rate in individual years was compared and the following conclusions were reached: The fertility rate is significantly (at the 1% level) lower in the transport centre in 1995 and 1996 (47%, 54%) as compared with the other centre the laboratory of which is used by the authors (57%, 61%). In 1997 the difference is no longer significant (63% x 64%). The difference in the pregnancy rate in centres for aspiration and embryo transfer in both centres is insignificant in the years of investigation. It may thus be stated that the oocyte transport does not have a negative influence on the effectiveness of the IVF + ET programme (same pregnancy rate) and thus makes it possible to include a greater and ever increasing number of patients in the IVF programme by making use of reserve capacities of embryological laboratories and thus reduce the total costs of this programme. PMID- 9750406 TI - [Sex life of pregnant women with cardiopathies]. AB - In a retrospective investigation comprising a group of 116 women with cardiopathies who delivered in 1989-1993. The author evaluated their sex life during the early puerperium, using the author's questionnaire "Sexual behavior and cardiac diseases" supplemented by a controlled interview. 41 women (35.3%) refused the interview on sexual behavior and did not complete the questionnaire. The analyzed group comprised 71 women with cardiac diseases. The maternal mortality rate in the investigated group was 0.86%, the perinatal mortality rate 0.0%, prematurity 3.4% and hypotrophy 5.2%. It was revealed that during the pregestation period 80% of the women with cardiac diseases has regular sex at least once a week and had an orgasm and satisfaction without any fears. With advancing pregnancy the libido declines, as well as the frequency of sexual intercourse, orgasm and sexual satisfaction and fear of sexual intercourse increases. Except during pregnancy and at its onset the spectrum of coital positions is varied. At the end of pregnancy lateral positions predominate. Sexual activity before pregnancy is usually initiated by both partners. At the end of pregnancy in first pregnancies it is the woman, in pluriparas it is usually the man. Interest in the sexual behavior of women with cardiac diseases on the part of cardiologists and obstetricians was minimal. In the author's opinion there is no reason to restrict sex life during pregnancy in women with cardiac diseases functional stage I and II (NYHA, 1964) unless there are obstetric or cardiological contraindications. PMID- 9750407 TI - [Use of condoms among the population of the Czech Republic: results of a national study]. AB - Based on an anonymous survey, using questionnaires, in a representative group of the population of the Czech Republic above 15 years of age (862 men and 857 women) the authors investigated the use of condoms at the onset of sex life and with a steady and casual sexual partner. From the results ensues that the use of condoms in our population is lower than in advanced western European countries and the USA. During the first intercourse it was used only by 19% men and 14% women, in particular those of younger age groups. In a steady partnership two thirds of the respondents have experience with a condom, however, only one fifth uses it regularly. The position is similar during intercourse also with casual sexual partners, though the respondents use it more frequently. However not even then one third of men and almost half the women do not use protection by condoms. The most important influence on the reported use of condoms is exerted by the factors of sex (men, probably to appear socially in the most favourable light report its more frequent use) and age (condoms are used more frequently by younger men and women). In the Czech population the effect of religion and education is not important in this respect. PMID- 9750408 TI - [Sarcoma of the uterus]. AB - Our experience with the treatment of uterine sarcoma is presented as a retrospective clinical study. The studied group is represented by 39 patients with the average age 58 years and an overall 5 year survival 36%. We recommend the abdominal hysterectomy with or without bilateral salpingooophorectomy as primary therapy. The importance of adjuvant radiotherapy and chemotherapy is discussed with a special attention to various histological types of sarcoma. PMID- 9750409 TI - [Intrapartum fetal pulse oximetry. A new method for the diagnosis of intrauterine fetal hypoxia]. PMID- 9750410 TI - [Misoprostol and induction of labor--an effective method of treating prolonged pregnancy]. PMID- 9750411 TI - [Study of sexual reactions in women]. PMID- 9750412 TI - [Use of highly purified FSH (Metrodin HP) in the management of ovarian hyperstimulation in the IVF/ET program--the first pregnancy and birth in the Czech Republic]. PMID- 9750413 TI - [Hormonal contraception and thromboembolic disease--II. Pathophysiologic basis]. PMID- 9750414 TI - [I. Free radicals and antioxidants in gynecology]. PMID- 9750415 TI - [II. Free radicals and antioxidants in obstetrics]. PMID- 9750416 TI - [40 years' since the founding of the Gynecologic Endocrinology Group in Prague Podoli]. PMID- 9750417 TI - [Analysis of maternal mortality in the Czech Republic in 1996]. PMID- 9750418 TI - [Conservative treatment of endometriosis]. PMID- 9750419 TI - [Primary cutaneous B-cell lymphoma: present views]. AB - Most lymphoproliferative B cell cutaneous lesions do represent primary lymphomas, not a secondary lymphomatous spread or pseudolymphoma* A description of the development of classification systems for primary cutaneous B lymphomas is given concerning especially some newly defined lesions: primary cutaneous marginal zone B cell lymphoma, primary cutaneous follicular centre cell lymphoma and large B cell lymphoma of the legs. Majority of primary cutaneous B lymphomas answer well the treatment and have a favourable prognosis. PMID- 9750420 TI - [Hodgkin's disease with epithelioid granulomatous reaction]. AB - Hodgkin's disease with a conspicuous presence of epithelioid-histiocytic elements was described as a particular subvariant of mixed type Hodgkin's disease. The authors presented four new cases. An additional increase of Langerhan's and interdigitating cells was observed in two cases, sarcoidosis-like granulomas with epithelioid and big multinucleated cells in the third. This female died and granulomas were found in several other organs. A prognostic relevance of this conspicuous reaction has not been clear for the time being. The mentioned patient deceased one year after the diagnosis had been settled, other three patients have been clinically well surviving 2-5 years. PMID- 9750421 TI - Desmoplastic ameloblastoma. AB - Two cases of desmoplastic form of ameloblastoma are reported. This tumor shows marked stromal desmoplasia and often scattered osteoplasia. The majority of stromal elements is represented by myofibroblasts. The epithelial component of this tumor is often scanty and different from epithelial islands of classic ameloblastoma. Pathological, immunohistochemical and radiological findings are described and the differential diagnosis of this tumor is discussed. PMID- 9750423 TI - [Ossification of pulmonary tissue]. AB - Two cases of pulmonary ossification found in postmortems were described. The first case, a 67-year-old stop-smoker with systemic hypertension died of a ruptured atherosclerotic aneurysm of the abdominal aorta. A well-defined subpleural mass (52 mm in max. dimension) was composed of monotonous mature trabecular osseous tissue with two focuses of cartilage up to 500 microns. There was no evidence of occluded blood supply and chronic passive congestion in the pulmonary parenchyma. A diagnosis of mesenchymoma (osteohamartoma) with the predominance of osseous tissue was made. The second case, a 72-year-old man non smoker died of urosepsis. There were no pulmonary symptoms in patient's previous history. A well-defined ovoid mass (25 mm in diameter) was subpleurally located. Microscopically, it was formed of small nodules of mature lamellar bone together with branching bony deposits of trabecular character. Moreover, there were multiple smaller focuses scattered in the distant pulmonary tissue having appearance of nodular ossification. The evidence of chronic thromboembolic pulmonary disease together with ossification in microscopy permits the possibility that the nodular ossification may develop from pulmonary scarring or hemorrhagies as a late consequence of the thromboembolic pulmonary disease. PMID- 9750422 TI - [Microstructure of subcutaneous lesions in juvenile hyaline fibromatosis]. AB - Juvenile hyaline fibromatosis is a rare autosomal recessive interstitial disease characterized by nodes and tumours of skin and soft tissues as well as by gingival hyperplasia. The authors described a case of 28-year-old male based on histopathological diagnosis. The patient was admitted to the hospital thrice in his life with the diagnosis of arthrogryphosis. Last time he presented with extensive secondary impetigo in extremities and pachydermia, polymalformation syndrome, multiple subcutaneous tumours, gingival hypertrophy, contractures, osteolytic lesions and positive family history. In histology, tumoriform lesions showed a structureless hyaline matrix often with chondroosseous metaplasia and calcium salts. More or less numerous cells in the matrix had a fibroblastoid appearance with eosinophilic cytoplasm, oval nuclei and frequently pericytoplasmic halo. Electron microscopy revealed dilated cisterns of rough endoplasmic reticulum and a hypertrophied Golgi apparatus. Particles representing calcium salts according to their density were rare. Immunohistochemistry of tumour cells showed vimentin, alpha-1-antichymotrypsin and alpha-1-antitrypsin. The findings concurred with the literature in which, nevertheless, the immunohistochemical picture were not mentioned. PMID- 9750424 TI - [Pseudoangiomatous hyperplasia of mammary stroma]. AB - Four cases of pseudoangiomatous hyperplasia of mammary stroma (PAH) were described (3 women and 1 man aged 15 to 42 years). Clinically in women, they presented as firm, nontender, movable nodules, 1 to 4 cm in diameter, diagnosed clinically usually as fibroadenoma. In the male patient, the lesion was tender, resembling gynecomastia. Microscopically, they were characterized by the presence of anastomosing, empty-looking, slit-shaped spaces irregularly lined by the spindle cells with small, uniformly ovoid nuclei, dispersed throughout the mammary stroma. Immunohistochemically, these cells were vimentin positive, factor VIII and CD31 negative. They did not express estrogen receptor, in one case there was a weak progesteron receptor positivity. Ultrastructurally, the lining cells were of a fibroblastic character. The pathogenesis of PAH is unclear, presumed to be of reactive nature by the authors, probably as an exuberant reaction of stromal cells to hormonal stimuli. In differential diagnosis, PAH should be distinguished from low-grade angiosarcoma, as well as from perilobular hemangioma, diffuse angiomatosis and vascular anomalies. PMID- 9750425 TI - [Over a half-century at the Charles University 1st Medical Faculty. 2. Pathology under Professor Sikl]. AB - Hlava Institute of Pathology was directed in a pleasant climate of broadminded indulgence. Sikl was easy to get in favour being personally attractive and imposing in work. He always sided with the youth and helped us even against authorities which made him more often laugh critically than treat with respect. Hlava Institute under Sikl became a model of progressing narrow specialization and modernizing. Sikl filled it with a programme of perspective development. Thus he prepared the whole discipline for cooperation which was useful for comparison of all findings with clinical picture. He made the biopsy independent and carried out its wide appreciation. He insisted upon the usage of big scale surveying histological cuts which he investigated thoroughly in detail. He did not live to see an explosion of methodological possibilities. Nevertheless, he prepared the structural diagnostics for an effective cooperation of all specialized methods. PMID- 9750426 TI - [Immunohistochemical study of inflammatory changes in the myocardium]. AB - A common histological investigation of the myocardium was compared with immunohistochemical testing of cardiomyocytes for fibrinogen and myoglobin in a 26-year-old man who died suddenly with a several days history of respiratory ways infection. A very discrete finding in hematoxylin eosin showed a slightly non purulent interstitial myocarditis whereas immunohistochemistry revealed a substantially bigger lesion of heart muscle fibres. Samples from different parts of myocardium showed disseminated or confluent lack of myoglobin and corresponding deposition of fibrinogen in injured cardiomyocytes. Myoglobin and fibrinogen appeared as a very appropriate combination of methods enabling to diagnose even a very minute injury of the heart muscle fibres. PMID- 9750427 TI - [Determination of the presence of traces of biological material using molecular genetics]. AB - Polymerase chain reaction is often used for molecular genetic human identification in forensic medicine today. In this study, the possibilities are demonstrated, which are provided by the method in such cases, where it is necessary to determine, if the biological traces are of animal or human origin. The method is rapid and well performed in a molecular genetic laboratory with standard equipment. Eleven samples of DNA isolated from different animals and birds were examined. In all cases, it was possible to distinguish these samples according to the results of reaction from simultaneously examined human DNA. PMID- 9750428 TI - [An outline for evaluating gunshot wounds from the International Council of the Red Cross]. AB - Paper summarized a concise evaluation of short wounds according to the Red Cross Classification published in Geneva. It is supposed to attract the forensic physician's attention due to informative purpose. PMID- 9750429 TI - [Study of diversity of the animal population in zonal climatic gradients using the transect method]. AB - Zonal trends of changes in animal population diversity on land are studied with the models of individual taxa and multispecies communities. Development of the transect method and use of different scale topoecological gradients make it possible to follow the succession of the species composition and topical preferences of individual species, as well as changes in their absolute and relative abundance, which lead to changes in the structure and pattern of functioning of the community as a whole. Modern methodological approaches concerning organization of complex studies on megatransects crossing several natural zones are analyzed. Methods of estimation of inter- and intrazonal changes in the animal population structure and diversity are discussed. PMID- 9750430 TI - [Strategy of preservation and use of biological resources in China]. AB - The current state of biological resources of China is characterized by the example of animal world resources. Dynamics of the protected territories, control over the world trade of animals, and methods of protection of animals in situ and ex situ are described. PMID- 9750431 TI - [Antelopes of China]. AB - A brief characterization of the distribution and ecology of six species of Chinese antelopes (Przewalski's gazelle, saiga, Tibetan antelope, four subspecies of Persian gazelle, Mongolian gazelle, and Tibetan zeren)is made, with special reference to the state of their population and protection in situ. PMID- 9750432 TI - [Current state and perspectives of the international cooperation for protection and use of the mongolian gazelle]. AB - The current status of the Mongolian gazelle population is described. The range of this species is located in the territories of China, Mongolia and Russia. Urgent tasks for investigation and conservation of this species within the framework of international cooperation are discussed. PMID- 9750433 TI - [Effect of rodents on reforestation in the moumtain regions near Beijing]. AB - From 1993 until 1995, the influence of rodents on reforestation in the mountain regions near Beijing was studied in order to estimate the pressure of rodents on the seed supply, search for effective ways to decrease the loss of seeds due to consumption by rodents, and increase seed germinating power. It was shown that acorns, apricot pits and nuts are almost entirely carried away from the soil surface by rodents, thus suggesting that rodents have a great impact on the seed supply. When the seeds were sowed at a depth of about 5 cm, many of them were not eaten by rodents; 39% of acrons and 18% of apricot pits germinated on the following year. We propose that deep sowing of seeds may effectively decrease the loss of seeds through consumption by rodents. PMID- 9750434 TI - [Amur tiger: path to the third millennium (experience of development of the "Strategy of preservation of the Amur tiger in Russia")]. AB - The development of a system of measures for conservation of the Amur tiger is summarized. In the middle of the 1990s, no marked reduction in the numbers of the Amur tiger took place in Russia; the animals were always present in most of the forest-covered area of the Primorskii and South Khababrovsk districts. However, new threats to the survival of the Amur tiger appeared. It is necessary to preserve the self-regulating population of the Amur tiger throughout its range. The key to solving this task is the identification of three functionally unequal zones in the southern Far East of Russia: a monolithic "tiger bastion" consisting of protected territories with different statuses, a zone of "stable compromise" (between the interests of tiger conservation and economic development of the territory), and a "tiger-free" zone, where the animals periodically migrate. The importance of Russian-Chinese cooperation in conservation of the Amur tiger has been stressed. A large international borderline reserve-cluster for this purpose has been proposed. PMID- 9750435 TI - [Sensitivity of human melanoma xenografts to nitrosoalkylurea antineoplastic agents]. AB - We studied the sensitivity of human melanoma (Bro strain) xenografts to drugs of the nitrosoalkylurea (NAU) class: nitrosomethylurea (NMM), karmustin (BCNU), nimustin (ACNU), nitrulin, and ADEKO. High antitumor activity of NAM was shown when the drugs were applied not only at the early, but also at the late stages of tumor progression (tumor mass 400 and 1200 mg, respectively). The therapeutic effect of the drugs was estimated with the use of criteria characterizing the kinetics of tumor regression, increased life span, and survival of treated animals. After early administration of the drugs (Day 4 after tumor transplantation), 67% and 50% of animals survive under the influence of nitrulin and ACNU, respectively, while the rate of tumor regression increased in the sequence nitrulin < karmustin < NMM < ACNU. After late administration (11 days after tumor transplantation), NMM was most effective at increasing survival (35% of survived animals by 35 days of observation), while the rate of tumor regression increased in the sequence ADEKO < NMM < karmustin < nitrulin < ACNU. PMID- 9750436 TI - [NO-Dependent mechanisms of adaptation to hypoxia]. AB - Studies of nitrogen oxide (NO)-dependent mechanisms of organism resistance to hypoxia demonstrate that (1) acute hypoxia induces NO hyperproduction in the brain and does not affect NO production in the liver; (2) adaptation to hypoxia decreases NO production in the liver and brain; and (3) adaptation to hypoxia prevents NO hyperproduction in the brain and enhances NO synthesis in the lever during acute hypoxia. An NO donor--dinytrosyl iron complexes (DCI, 200 micrograms/kg, single intravenous (i.v.) introduction)--decreases animal resistance to acute hypoxia by 30%, while introduction of an NO synthase inhibitor--N- nitro-L-arginine (NNA, 50 micrograms/kg, single intraperitoneal (i.p.) introduction)--and an NO trap--diethyldithiocarbamate (DETC, 200 mg/kg, single i.p. introduction)--increases the resistance 1.3 and 2 times, respectively. Adaptation to hypoxia is realized against a background of accumulation of heat shock proteins HSP70 in the liver and brain. Course treatment with DCI reproduces the antihypoxic effect of adaptation to hypoxia. Course treatment with NNA during adaptation to hypoxia prevents both accumulation of HSP70 and development of the antihypoxic effect. Hence, No and NO-dependent activation of HSP70 synthesis play an important role in adaptation to hypoxia. PMID- 9750438 TI - Which type of diffuse emphysema is adequately contracted by the Nd:YAG laser. An ex-vivo experiment. AB - Diffusely emphysematous lungs are not always effectively contracted by laser therapy; however, which type of diffuse emphysema that responds to laser therapy remains unclear. We macroscopically and histopathologically examined human lung tissue, which was resected from patients with carcinoma, after irradiation with an Nd:YAG laser. Forty-six lung lobes were irradiated with a non-contact mode Nd:YAG laser at a power setting 15 watts. Macroscopically, twenty samples of normal lungs revealed moderate contraction, fourteen samples of predominantly centrilobular diffuse emphysema showed significant contraction, and eight samples of predominantly panlobular diffuse emphysema with a slight elastic network showed slight contraction. Histopathologically, the normal lungs showed amorphous change of the collagen and severely contracted elastic fibers (amorphous degeneration) at the pleura and some parenchymal coagulation; the predominantly centrilobular diffuse emphysema showed contraction of elastic fibers and collagen (coagulative degeneration) in the pleura and adequate contraction of the elastic fibers in the parenchyma and the predominantly panlobular diffuse emphysema showed only slight coagulation of the visceral pleura and very little coagulation of the parenchyma. On ex-vivo lung, panlobular emphysema was inadequately contracted by laser therapy, due to elastic recoil. Centrilobular emphysema responded to laser treatment, due to the severe contraction of the elastic fibers. PMID- 9750437 TI - [Radionuclides in the Kara sea bottom sediments]. AB - The contents and distribution of natural (214Pb, 214Bi, 228Ac, and 208Tl) and technogenic (90Sr, 137Cs, and 60Co) radionuclides in the Kara Sea bottom sediments was analyzed by using the materials collected by R/V "Pomor" (Murmansk Marine Biological Institute). In 1994, no high (critical) concentrations of technogenic radioisotopes were found in the sea sediments, while the spots (regions) of elevated 137Cs content found in 1984 were not confirmed in 1994. It was proposed that the main sources of entry of technogenic radionuclides in the sea sediments in the last years are the flow of the Ob'-Yenisei waters and container burials of radioactive waste in the sea, which appeared to have been markedly corroded. The latter is confirmed by detection in some places of 60Co, which was not previously found in the sediments. PMID- 9750440 TI - Aortic dissection extending from ductus diverticulum aneurysm. AB - We report the case of a 28-year-old woman with type B chronic aortic dissection extending from a ductus diverticulum aneurysm. The patient was successfully treated by interposition of a Hemashield 26-mm woven Dacron graft with a left posterolateral thoracotomy approach. Circulatory arrest through profound hypothermia and short retrograde cerebral circulation were employed during proximal anastomosis of the graft. PMID- 9750439 TI - Left atrial ball thrombi without mitral valve disease treated by surgical removal. AB - We describe two patients with free-floating left atrial ball thrombi with no evidence of cardiac disease except atrial fibrillation. One patient had experienced an embolic stroke, and the second patient had a history of stroke and peripheral thromboembolism. In each patient, the thrombi, which were not visible on transthoracic echocardiography, were detected by transesophageal echocardiography. Each patient underwent successful surgical removal using cardiopulmonary bypass. PMID- 9750441 TI - New left main trunk reconstruction technique using a flap of the main pulmonary arterial wall. AB - A new technique is described for ostiumplasty of the left main coronary artery using a flap of the main pulmonary arterial wall. Venous or pericardial patches are conventionally used to enlarge coronary ostial lesions, we used a flap of the pulmonary arterial wall with expectation that it is viable over the long term. This technique seems to surpass conventional coronary artery bypass grafting. PMID- 9750442 TI - [Evaluation of cerebral circulation during cardiopulmonary bypass using near infrared spectroscopy]. AB - Cerebral oxygenation level during cardiopulmonary bypass (CPB) was measured using near-infrared spectroscopy as a monitor of cerebral circulation in 30 patients. Six adult cases with thoracic aortic aneurysm were operated on using selective cerebral perfusion (SCP). CPB was established under moderate hypothermic temperature in 9 adult cases (hypothermic group, lowest blood temperature during CPB; 25 degrees C) and under normothermic temperature in 9 adult cases (normothermic group, lowest blood temperature during CPB; 34 degrees C). In congenital cases (n = 6), CPB was established under moderate hypothermic temperature (congenital group, lowest blood temperature; 25 degrees C). The oxyhemoglobin (HbO2) level showed a significant positive correlation with cerebral blood flow during SCP (r = 0.715). There was no significant correlation between SjO2 and HbO2 in the SCP group. The HbO2 levels in the hypothermic group after 30 and 60 min, from the initiation of CPB and 30 min. before the weaning of CPB were significant lower than the control level (p < 0.05). HbO2 levels in the congenital group after 0, 30 and 60 min. from the initiation of CPB and 30 min. before the weaning of CPB were significantly lower than the control level (p < 0.01). The deoxyhemoglobin (HbR) level in the hypothermic group after 30 and 60 from the initiation of CPB and 30 min. before the wearing of CPB were significantly higher than the pre level (p < 0.05). The mixed venous saturation (SvO2) in the normothermic group showed significant lower levels than those in the hypothermic group (p < 0.01). However, there was no significant difference in HbO2 levels between the two groups. In conclusion, these results suggest that near-infrared spectroscopy may be a noninvasive and useful technique for the cerebral circulation monitoring during CPB. PMID- 9750443 TI - [Open distal anastomosis or aortic balloon occlusion technique during complete aortic arch replacement]. AB - The technique of open distal anastomosis or application of aortic balloon occlusion catheter designed to occlude the descending thoracic aorta have been used in 33 and 19 patients, respectively, to control bleeding during the procedure of distal anastomosis for complete aortic arch replacement with a prosthetic graft. These two techniques allowed us a simple approach to the lesion and the avoidance of clamp injury to the fragile aortic tissue. Open distal anastomosis was applied for 91% patients of operated aortic dissection and all emergent cases, it's duration ranged from 10 to 110 minutes with an average of 58 minutes under 18.2 degrees C of lowest esophageal temperature. On the other hand, aortic occlusion balloon was inserted for mainly true aortic aneurysm patients without an emergency, and helped to maintain the perfusion pressure on a lower part of body around 50 mmHg by the 1550 ml/min in an average of perfusion flow femoral artery under 21.2 degrees C of temperature. The difference of postoperative renal and liver function evaluated by serum enzyme levels of total bilirubin, GOT, GPT, LDH, creatinine and BUN did not reach to statistical significance between the patients using open distal anastomosis and balloon occlusion, however, the incidence of postoperative complication including either renal, liver dysfunction, abdominal problem or paraplegia was significantly higher in the patient group with open distal technique. Either open distal anastomosis or aortic balloon occlusion technique would be appropriately selected according to the patient's characteristics or the condition of aortic disease to be operated. PMID- 9750444 TI - [The effect of intraoperative high-dose tranexamic acid on blood loss after operation for acute aortic dissection]. AB - The effect of high dose tranexamic acid on blood loss after operations for acute aortic dissection was evaluated. Twenty-eight patients undergoing emergent operations for acute aortic dissection were studied. There were two groups, group T with 13 patients (group T) who were given 7 g of tranexamic acid after induction of anesthesia and 3 g of it after CPB and group C with 15 patients who did not receive tranexamic acid. There was a tendency that group T had less bleeding during operation and after operation (559.6 +/- 865.8 ml in group T and 805.8 +/- 442.9 ml in group C, 1719.2 +/- 1008.7 ml in group T and 3547.7 +/- 4580.1 ml in group C, respectively), but there was no significant difference between two groups. The removal of drainage tubes after operation was significantly earlier in group T (5.0 +/- 2.3 post operative day in group T and 8.1 +/- 5.2 post operative day in group C; p < 0.05). FDP and D-dimer level as measures of fibrinolytic activity were elevated at pre- and postoperative period in both groups, but they tended to be lower in group T at postoperative period. One patient required reexploration because of excessive bleeding and no mediastinal infection was reported in group T, whereas 4 patients underwent reexploration and 2 patients developed mediastinitis in group C. There were 5 hospital death (33.3%) in group C and 2 (15.4%) in group T. High dose of tranexamic acid seems to control fibrinolytic activity, thereby reducing blood loss and requirements, which may contribute to lower morbidity and mortality in operations for acute aortic dissection. PMID- 9750445 TI - [Effects of myocardial protection with ryanodine, measured with the intracellular calcium fluprescent indicator Fura-2]. AB - The effects of Ryanodine, an inhibitor of salcoplasmic reticulum function, was investigated in isolated hearts of the Wistar rat strain. The cytosolic calcium was measured with the intracellular Ca2+ fluorescent indicator Fura-2. After 3 minutes perfusion of various cardioplegic solution which were added potassium (the concentration; 20 mmol/L) and four Ryanodine groups (1, 4, 10, and 20 nmol/L), these were obtained cardiac arrest. The arrested hearts were kept at 37 degrees C (normothermia) measuring hemodynamic studies. Hemodynamic parameters were heart rate, LVDP, LV dp/dt, coronary flow, and the intracellular calcium fluorescents which were calculated intracellular Ca2+. The significant difference was not noted in LVDP, but comparable improvement was achieved with Ryanodine groups (p < 0.05; 20 nM vs 0 nM). Other cardiac functions were likely same as above. The cytosolic calcium concentration of Ryanodine groups was depressed during cardiac arrest, it was 97.4 +/- 17.2% at the end of cardiac arrest, and it was slowly increased to 161.9 +/- 46.9% after 40 minutes reperfusion. On the other hand, that of the control group was higher than Ryanodine groups at every measuring points about two times. There was significant difference between both groups (p < 0.01). Concequently these phenomena caused that Ca2+ handling in the salcoplasmic reticulum were supressed by Ryanodine and the contractile function was recovered for that reason. PMID- 9750446 TI - [Cardiac hemangioma of the right ventricle--report of a case]. AB - A 74-year-old male was admitted to our hospital with a diagnosis of intra-cardiac tumor on echochardiography, CT, and MRI. The tumor was located on the free wall of the right ventricle and protruded into the outflow tract. The surgical excision of the tumor was performed under cardiopulmonary bypass. The size of the tumor was 5 x 5 x 4 cm. Histological examination disclosed cavenous hemangioma. The post opertive course was uneventful. This is the forth case reported in Japan and the fifteenth case in the world. PMID- 9750447 TI - [Multiple lungs tumors found 17 years after hysterectomy--a case of benign metastasizing leiomyoma]. AB - The patient is a 50-year-old female who presented with cough. A chest X-ray showed bilateral multiple modular shadows. Even though a diagnosis of metastatic lung tumors was strongly suspected no primary lesion could be detected on close examination. Therefore, an exploratory thoracotomy, and histopathological examination of a specimen obtained from a resected tumor were carried out. The resected tumor revealed pathological proliferation of spindle cells without high cellularity or mitosis with nuclear atypism. The patient had a history of surgery for uterine leiomyoma at the age of 33. The histopathological characteristics of the resected lung tumors were remarkably similar to those of the previously resected uterine leiomyoma. In addition, the serum levels of sex hormones, including LH, measured after lung surgery, were within normal range suggesting the possibility of the lung tumors being metastatic. The residual lung tumors did not show any increase in size 26 months after surgery. Therefore, these lung tumors could be considered as lung metastases from a low grade malignant uterine leiomyoma (benign metastasizing leiomyoma). PMID- 9750448 TI - [Successful repair of critical aortic stenosis with coarctation on the first day of life]. AB - Successful open aortic valvotomy and end-to-end anastomosis were performed to the patient with critical aortic stenosis and CoA on the first day of life. A fetus was diagnosed as aortic stenosis and coarctation by fetal echocardiography at 29 weeks of gestation. The mother was transferred to our hospital at the onset of labor and delivered vaginally at 40 weeks of gestation. Soon after the birth, the newborn, birth weight 2630 gram, female, underwent echocardiography by pediatric cardiologists which demonstrated the aortic orifice of 5.1 mm in diameter and thickened cusps with poor mobility. Transaortic pressure gradient measured by Doppler echocardiography was 111 mmHg and the left ventricular wall motion was hyperdynamic without any signs of endocardial fibroelastosis. Prostaglandin E1 administration was started to maintain systemic circulation and the hemodynamic status has been stable before surgery. On her first day of life, the operation was performed using cardiopulmonary bypass with moderate hypothermia. Isolated cerebral and myocardial perfusion technique was applied during the repair of coarctation of the aorta. The open aortic valvotomy with resection of myoxomatous nodules on leaflet edges effectively released pressure gradient across the aortic valve without regurgitation. Post-operative course was uneventful and she discharged at 28th day after surgery. We conclude early diagnosis including fetal echocardiography and early repair would lead the better surgical outcome to the neonates with critical aortic stenosis. PMID- 9750449 TI - [Percutaneous transcatheter coil embolization of the patent ductus arteriosus for elderly patient with left ventricular disfunction]. AB - We performed the coil embolization for 64-year-old male with patent ductus arteriosus and left ventricular dysfunction. We used "snare method" and "cross catheter technique" delivering one coil transvenously and one coil transarterially. Echocardiograms at 7 month after the procedure demonstrated complete occlusion of the ductus, though small shunt was detected at discharge. PMID- 9750450 TI - [A mitral valve reconstruction of infective endocarditis with brain abscess and intracranial mycotic aneurysm]. AB - A case is reported of a brain abscess and an intracranial mycotic aneurysm associated with infective endocarditis caused by streptococcus intermedius. A 60 year-old man with a history of fever presented aphasia and right hemiparesis. A computed tomographic scan of the head revealed a low-density area with ring enhancement in the left parietal lobe consistent with a brain abscess. An angiography demonstrated an aneurysm on the distal branch of the middle cerebral artery compatible with a mycotic aneurysm. Doppler echo cardiography showed severe mitral regurgitation by chordal ruptures. The brain abscess and intracranial mycotic aneurysm were resolved under appropriate antibiotic therapy for eight weeks. Then, the mitral valve was reconstructed by replacement of the chordae tendineae with expanded polytetrafloroethylene suture and annuloplasty. The patient had no neurologic deficit except for paresthesia in the right hand, and had no mitral regurgitation at discharge. PMID- 9750451 TI - [Bronchial carcinoma: Paclitaxel combinations are administered at an ever earlier stage of the disease. 34th Annual Meeting of the American Society of Clinical Oncology (ASCO). Los Angeles, 16-19 May 1998]. PMID- 9750452 TI - [5th International Symposium, Bad Lippspringe. "Management of Asthma and Allergy". Bad Lippspringe, 16 May 1998]. PMID- 9750453 TI - [Outcomes of 50 leukemia patients who received bone marrow transplants from unrelated donors]. AB - Fifty leukemia patients were given bone marrow transplants (BMTs) from unrelated donors at Meitetsu Hospital. We studied the outcomes of their transplants from two perspectives: leukemia disease stage and acute graft versus host disease (GVHD). The probability of disease-free survival for standard-risk, high-risk, and super-high risk patients was 65%, 29%, and 8%, respectively. The main causes of death were septicemia, cardiac and renal failure, and relapse of leukemia in the high- and super-high risk patients, and grade III-IV acute GVHD in the standard-risk patients. The incidence of grade II-IV and grade III-IV acute GVHD was 32% and 17%, respectively. All 7 patients in whom grade III-IV severe acute GVHD developed died. We conclude that better control of acute GVHD and treatment of early stage complications are clearly important to improving the outcome of BMTs from unrelated donors, especially for high and super-high risk patients. PMID- 9750454 TI - [Prognostic significance of CD7 expression in adult acute myeloid leukemia]. AB - To evaluate the prognostic significance of CD7 expression in de novo acute myeloid leukemia (AML), we studied 63 patients with AML who had been admitted to our hospital between September 1989 and January 1996. Even of the patients were later eliminated from the study (9 due to insufficient surface marker analyses, and 2 due to early death). The remaining 52 patients (median age: 42.5 years) were evaluated for morphologic subtype, immunophenotypic classification, complete remission (CR), disease-free survival (DFS) and overall survival (OS). All 52 patients were grouped by the French-American-British classification system: 10 as M1, 16 as M2, 11 as M3, 8 as M4, 5 as M5, and 2 as M6. Ten of the patients expressed CD7 on their leukemia cells (positive rate > or = 25) and were classified as CD7(+)AML, with morphological subtypes as follows: 3 as M1, 6 as M2, and 1 as M3. Thirty-three of the 42 patients with CD7 + AML (78.6%) and 6 of the 10 patients with CD7 + AML (40%) achieved CR. DFS and OS rates for the patients with CD7(+)AML were 22.1% and 35.4%, respectively; those for the CD7(+)AML patients were 53.3% and 44.4%, respectively. No significant differences in gender hematological findings, clinical manifestations such as hepatosplenomegaly, lymphadenopathy, or incidence of central nervous system involvement, CR rate, and DFS distinguished patients with CD7(+)AML from those with CD7(+)AML. These suggest that CD7 expression is unlikely to be a prognostic factor in AML. PMID- 9750455 TI - [Effective pentostatin-based treatment of adult T cell leukemia in a patient with severe arthritis]. AB - We report a 48-year-old woman with adult T-cell leukemia who had refractory arthralgia, intense headaches, and fever. Leukemic cell infiltration of the cerebrospinal fluid was detected but no other acute signs were observed. Abnormal lymphocytes with lobulated nuclei were found in the synovial fluid, and a histologic examination revealed proliferation into the synovium. Because combination chemotherapy did not elicit a favorable response, the patient was treated with a pentostatin bolus injection. The articular symptoms disappeared and complete remission was obtained. Six months later, she experienced arthralgia again together with a gradual increase of abnormal lymphocytes in peripheral blood. Sixteen months later, the patient was given pentostatin and achieved a complete remission again. She is still free from relapse without further therapy after 36 months, and her articular symptoms have not returned either. There were no adverse effects due to pentostatin. The patient's serum IL-6 level was elevated, suggesting that IL-6 may play a role in arthropathy. PMID- 9750456 TI - [Polyclonal B-cell lymphocytosis with clinical and hematological features resembling hairy cell leukemia]. AB - A 49-year-old man was admitted to our hospital for investigation of splenomegaly and lymphocytosis. He had no significant past history and was not a smoker. Physical examination revealed massive splenomegaly and no palpable superficial lymph nodes. Hematological examination showed a hemoglobin concentration of 10.5g/dl, a platelet count of 9.8 x 10(4)/microliter, and a leukocyte count of 21.2 x 10(3)/microliter with 70% abnormal lymphocytes. In May-Giemsa stained blood films, the abnormal lymphocytes had round nuclei, abundant, pale cytoplasm, and slightly serrated edges. Phase-contrast microscopic and scanning electron microscopic examinations revealed many long surface villi. Tartrate-resistant acid phosphatase activity in these cells was negative. The abnormal lymphocytes had a CD5-, CD10-, CD11a+, CD11c+, CD19+, CD20+, CD22+ phenotype. These features were similar to those described for a variant form of hairy cell leukemia (HCL Japanese variant). However, studies of Ig gene rearrangement and expression of sIg revealed a polyclonal proliferation of B cells. On the basis of these findings, this case was diagnosed as hairy B-cell lymphoproliferative disorder, a recently described condition characterized by polyclonal B-cell lymphocytosis and features resembling HCL-Japanese variant. Serological assays for antibodies against Epstein-Barr virus suggested a past infection. Splenectomy alleviated the anemia and thrombocytopenia, but not the lymphocytosis. PMID- 9750457 TI - [Successful treatment with idarubicin in a pediatric case of t(8;21) acute myelogenous leukemia with additional chromosomal abnormalities at relapse]. AB - A 9-year old boy was admitted to our hospital due to a relapse of acute myelogenous leukemia (AML). A chromosomal analysis at the time of relapse revealed abnormalities in addition to 45, X,-Y, t(8;21) (q22;q22) when AML was first diagnosed. The patient was given granulocyte-colony stimulating factor (G CSF), cytosine arabinoside (Ara-C) and aclarubicin (CAG therapy), but this treatment regimen was not effective. He was next treated with G-CSF (started 3 days prior to the administration of anticancer drugs), Ara-C, (200 mg/mm2 for 7 days), Etoposide (VP.16, 150 mg/mm2 for 5 days) and Idarubicin (8 mg/mm2 for 5 days) according to the modified Japan Cooperative Protocol ANLL 91 for children. Although his condition had been septic and he had experienced renal and respiratory failure, he achieved a complete remission after 140 days without additional therapy. The patient returned to a condition of health and received a bone marrow transplant from an unrelated donor. We concluded that this treatment regimen is effective for the relapse of AML in children. PMID- 9750458 TI - [Plasma cell leukemia (IgG kappa) presenting bilateral neurosensory hearing loss and left sixth cranial nerve plasy]. AB - A 30-year-old man who had been given a diagnosis of IgG-kappa multiple myeloma by another hospital and treated with melphalan, prednisone, and cyclophosphamide 6 months earlier, was admitted to our hospitaly in July 1994 because of progressively impaired hearing in both ears, vertigo, and worsening fatigue. Peripheral blood examination showed a white blood cell count 25,000/microliter, with 77.5% atypical plasma cells. Examination at the time of hospitalization also revealed retinal hemorrhages and serum hyperviscosity. The diagnosis was plasma cell leukemia with hyperviscosity syndrome. Subsequent treatment consisted of vincristine, doxorubicine, and prednisone and repeated plasmapheresis. This resulted in a partial response and a reduction of serum viscosity but no reversal of hearing loss. One month after admission, left sixth cranial nerve plasy was demonstrated. Cranial computed tomography studies disclosed a tumoral mass in the sphenoid sinus. The patient received local radiotherapy and intensive chemotherapy, but exhibited no notable alleviation of his cranial nerve palsy. He died of septicemia and progressive disease in August 1994. This case was rare in that it involved plasma cell leukemia and bilateral neurosensory hearing loss associated with serum hyperviscosity and sixth cranial nerve plasy due to plasmacytoma within the sphenoid sinus. PMID- 9750459 TI - [Gamma-heavy chain disease associated with MALT lymphoma of the duodenum]. AB - We report a case of a 63-year-old woman with gamma heavy chain disease (HCD) associated with mucosa-associated lymphoid tissue (MALT) lymphoma of the duodenum. She was suffering from drug-resistant tonsillitis with high fever. Examination on admission showed leukocytopenia and thrombocytopenia. Bone marrow aspirate revealed granulocytosis and a hypocellular marrow with no increase in plasma cells or atypical lymphocytes. Serum electrophoresis disclosed, in addition to hypogamma-globulinemia, an abnormal band due to the presence of gamma HCD protein. This abnormal protein was a molecular weight of approximately 40 kd as determined by Western blots technique, and belonged to the IgG1 subclass as determined by ELISA with monoclonal antibodies against IgG. An endoscopic examination of the patient's duodenum found a small tumorous lesion, which was confirmed pathologically to be MALT lymphoma. HCD is known to be associated with lymphoproliferative diseases. In this case, gamma HCD had developed as a secondary complication of MALT lymphoma. gamma HCD associated with MALT lymphoma of the duodenum is rare in the literature. PMID- 9750460 TI - [Acute transformation of chronic myelomonocytic leukemia with t(1;3) (p36;q21) abnormality]. AB - A 66-year-old man was given a peripheral blood test because of low grade fever. Leukocytosis was detected, and the blood and bone marrow findings were consistent with those of chronic myelomonocytic leukemia. Three months later the hematological findings were: WBC 58,800/microliter (19% blastoid cells, 22% monocytes), Hb 9.0 g/dl, and a platelet count of 116 x 10(4)/microliters. A bone marrow examination revealed the presence of 52.6% blastoid cells and dysmegakaryocytopoiesis, including micromegakaryocytes. Serum and urinary lysozyme levels were elevated. Karyotypic analysis detected t(1; 3) (p36;q21), but not major bcr/abl mRNA. The patient was given a diagnosis of acute transformation of chronic myelomonocytic leukemia. Despite treatment, he died about 3 months later. t(1;3) is occasionally observed in cases of myelodysplastic syndrome (MDS) and leukemia. Patients with t(1;3) often exhibit dysmegakaryocytopoiesis; furthermore, acute leukemia develops more readily in those who also have MDS. Cases of long-term survival are rare. PMID- 9750461 TI - [Successful emergency operation for massive hemorrhage due to jejunal angiodysplasia after intensive chemotherapy in a patient with refractory anemia with excess of blasts]. AB - A 59-year-old man was referred to our hospital because of pancytopenia. Peripheral blood examination showed a WBC of 1,500/microliters with 2% blasts, Hb 8.1 g/dl and a platelet count of 4.1 x 10(4)/microliters. A bone marrow aspiration revealed hyperplasia with proliferation of blasts (15.7%) and myelodysplasia. Chromosome analysis revealed multiple aberrations, including -5, 7, +8. The patient was given a diagnosis of refractory anemia with excess of blasts (RAEB) and treated with combination chemotherapy. Agranulocytosis and high fever remained after chemotherapy, and abdominal pain and diarrhea developed. An abdominal X-ray film and computed tomography scan demonstrated dilated small bowel, thickness of the bowel wall, and ascites. A diagnosis of neutropenic enterocolitis was given. During the WBC recovery period from nadir, massive hematochezia developed in the patient. Angiography detected the leakage of contrast medium from a peripheral region of the first jejunal artery into the jejunal lumen. A partial resection of the jejunum was thus performed, and a histological examination revealed the presence of irregularly dilated blood vessels in the submucosal layer. These findings were consistent with the features of angiodysplasia, and indicate that angiodysplasia should be considered one cause of intestinal hemorrhage in elderly patients during intensive chemotherapy. PMID- 9750462 TI - [Leptin in persons with simple obesity]. AB - Obesity is a disease with distinct genetic determination and its phenotype is defined by the still unknown number of genes whose expression can be influenced by environmental factors. Several years ago, "obesity gene" was isolated in animals. This gene, coding protein which consists of 165 amino acids, is called leptin. Leptin is supposed to be a key substance controlling homeostasis of body weight and energy balance; it is produced by adipocytes and its value correlates highly significantly with anthropometric parameters that characterize physical constitution and amount of subcutaneous fatty tissue. The obese individuals often display hyperleptinemia which is frequently caused by a postreceptor disorder; sporadically, a different leptin structure or hypoleptinemia (caused by genetic anomaly) are reported. It is supposed that either absolute or relative leptin deficiency in obese persons are associated with causal obesity (e.g. appetite stimulation). Leptinemia values correlate with percentage of subcutaneous fatty tissue, insulinemia and sometimes with glycemia. In our study we examined 200 probands, patients of the Metabolic and Diabetologic Out-Patient Department, Hospital in Sternberk. A very close correlation between the amount of subcutaneous fatty tissue (measured by a caliper in 10 skinfolds) and the leptine serum concentration was found. The values of leptinemia in men of normal constitution ranged within 1-11 ng/ml, non-obese women had 3-4 times higher values. Leptinemia in some obese individuals reached up to 70 ng/ml. However, the currently calculated and reported parameters of physical constitution (BMI, WHR, Grant index) did not correlate significantly with leptinemia. Similarly, biochemical parameters considered as general markers of insulin resistance (often associated with obesity) did not correlate significantly with leptinemia. This finding indicates that some calculated parameters, quantifying and gualifying physical constitution, may be ambiguous and leptinemia was found to give more detailed information about the amount of subcutaneous fatty tissue than WHR or BMI. An accidental finding was an important positive correlation between myoglobin concentration and creatinemia. At monitoring the effect of hypolipidemic agents we use the myoglobin examination and therefore we consider this correlation to be very important and every physician performing this analysis should be informed about it. The present study thus confirmed that a more accurate quantification of subcutaneous fatty tissue is required. On the other hand, we believe that examination of leptinemia can contribute significantly to stratification of patients into risk groups (with respect to clinical, economic and time differentiation) and subsequently to the treatment of these patients. In future, criteria for quantification of leptinemia and leptine resistance should be defined precisely. PMID- 9750463 TI - [The effects of nicotine on leptin levels in patients with android obesity]. AB - In insulin resistant subjects with android obesity the leptin levels are, as compared with non-obese subjects, elevated in proportion to their BMI, WHR and their percentage of body fat. Generally independent on obesity, leptin levels are significantly higher in women than in men as in women the percentage of adipose tissue is higher. After administration of 2 mg nicotine in Nicorette chewing gum to 36 android obese non-smokers the elevated baseline values of leptin did not change and thus the observation that cigarettes suppress hunger or that smoking promotes weight reduction is untrue or else this effect is not mediated by nicotine stimulation of leptin secretion or formation in adipose tissue, leptin being the adipose tissue hormone which controls food intake, the sensation of satiety and via neuropeptide Y also other hypothalamic functions such as muscular and sexual activity, gonadoliberin output, thermoregulation etc. Leptin thus offers no alibi to smokers. Conversely smoking in android obese hyperinsulinaemic hyperleptinaemic subjects with syndrome X (5H) potentiates significantly the risk of cardiovascular death. PMID- 9750464 TI - [The metabolic syndrome, its heredity, methods of detection and clinical significance]. AB - BACKGROUND: The metabolic syndrome is a common denominator of a number of risk factors which are associated with type 2 diabetes and serious cardiovascular complications. The symptoms develop on the background of insulin resistance and are associated with hyperinsulinism. They are not manifestations of ageing, but are developing probably on a predisposed background already in young age. METHODS AND RESULTS: In the families of 39 probands with symptoms of the metabolic syndrome (MS) there were 58 offspring, mean age. 26.4 +/- 8.7 years. They were compared with 46 controls matched for age, BMI and sex without a family-history of metabolic diseases. The offspring from the families of probands with MS differed in particular by a lower HDL cholesterol level (1.38 mmol/l +/- 0.31 vs. 1.72 mmol/l 1-0.53, p < 0.001), a higher blood sugar level during the glucose tolerance test (p < 0.01) and a higher level of apolipoprotein B (Apo B, p < 0.01). The stimulated insulin concentration, the IRI sums were highly significantly raised (S IRI 240.9 microU/ml +/- 141 microU/ml vs. 177.1 microU/ml +/- 77.9 microU/ml, p < 0.01), similarly as the C peptide concentrations (S C peptide 7.61 pmol/ml +/- 2.84 pmol/ml vs. 5.02 pmol/ml +/- 1.49 pmol/ml, p < 0.001) which correlated mutually. The great majority of offspring came from parents with hypertension and hyperlipoproteinaemia resp., the rest from diabetic families and families with IHD. In offspring of families of hypertonics there was a lower ratio of linoleic acid in serum phospholipids, contrary to complement (= the other members of the group), in offspring of subjects with hyperlipoproteinaemia there were higher fibrinogen and uric acid levels. The offspring of diabetics had higher mean BMI values, in families of patients with myocardial infarction higher C peptide levels were found. CONCLUSIONS: Already at post-adolescent age there are convincing signs which suggest the possible development of metabolic syndrome in predisposed families, as ensues from a comparison with controls: raised IRI, C peptide and some MS component levels. Linoleic acid as well as fibrinogen, BMI and C peptide concentrations which differed in offspring with different types of family-history from the complement are also related directly or indirectly with hyperinsulinaemia. PMID- 9750465 TI - [Effect of a revascularization procedure on functioning of the hibernating myocardium]. AB - Persistent abnormalities of left ventricular wall motion in patients with chronic coronary artery disease can be reversed by successful coronary artery bypass surgery or coronary angioplasty. The identification of hibernating myocardium has therefore important therapeutic and prognostic implications. Echocardiography with low dose dobutamine infusion can detect viable myocardium in patients with chronic left ventricular dysfunction. We have studied 32 patients with angiographically confirmed coronary artery disease and left ventricular dysfunction. The effect of dobutamine infusion (5 micrograms/kg/min. followed by 10 micrograms/kg/min) on left ventricular function was evaluated before coronary revascularization (PTCA, n = 20); CABGm b = 12) and compared with early period after revascularization (1-7 days) and 6-7 months later. Before revascularization 226 segments were hypokinetic, akinetic or dyskinetic, improvement was observed in 122 segments during dobutamine echocardiography. Mean (+/- SD) segment score indexes were 2.50 +/- 0.17 at baseline, 2.04 +/- 0.19 (p < 0.001) after dobutamine infusion, 2.24 +/- 0.25 (p < 0.001) early after and 1.87 +/- 0.25 (p < 0.001) late after revascularization. Sensitivity of dobutamine infusion to predict improvement early and late after revascularization was 84% and 85% respectively, specificity was 85% and 90% respectively. Echocardiography during low dose dobutamine infusion is save and accurate method for identifying of hibernating myocardium and predicts early and late reversibility of wall motion in selected patients with coronary artery disease and chronic left ventricular dysfunction. PMID- 9750466 TI - [What are the trends in cardiovascular pharmacotherapy in recent years in patients after myocardial infarct?]. AB - The authors compared the status of pharmacotherapy in 587 patients with myocardial infarction in 1991 according to the MONICA study, in hospitals of six districts in the Czech Republic, age group 25-64 years with data for 1995 assembled by means of questionnaires in 749 patients of the same age group, who were after myocardial infarction in selected spas. Thrombolytic treatment proved quite inadequate in 1988-1993 in six districts of the Czech Republic, although it rose slightly from mere 2.9% in 1988 to 11.1% in 1993. In 1995 25.4% patients with acute myocardial infarction were treated by thrombolytic treatment. Conversely anti-platelet treatment was very satisfactory in 1991 as well as in 1995. Promising was also the increase in treatment with beta-blockers and ACE inhibitors in 1995 as compared with 1991. Also the percentage of patients treated by hypolipidaemic treatment increased in 1995. This work indicates the favourable trends in pharmacotherapy of patients with acute myocardial infarction in the acute stage as well as in patients after myocardial infarction. PMID- 9750467 TI - [Pulsed doses of calcitriol in the treatment of secondary hyperparathyroidism in patients on hemodialysis]. AB - Administration of pulse doses of calcitriol is a better way of conservative treatment of secondary hyperparathyroidism (2HPT), making use of the direct suppression of parathormone (PTH) secretion. In a group of 29 haemodialyzed patients the authors evaluated during a six-month follow-up the effect of intravenous Calcijex in 12 and of oral Rocaltrol in 8 subjects. In responders of the calcijex group the PTH level declined by 67.6%, the mean baseline PTH value being 787.8 pg/ml, as compared with non-responders where the decline of PTH at the end of the investigation was 7.5%, the baseline PTH being 1296.4 pg/ml. The difference was significant (p < 0.05). In patients treated with Rocaltrol the therapeutic effect was apparent also in subjects with a lower baseline PTH. An associated phenomenon of treatment are as a rule parallel changes of kALP and ACP levels with those of PTH. It was however revealed that the drop of serum activities can occur also without a concurrent drop of PTH which indicates a dissociation between the level of bone metabolism and PTH secretion. The therapeutic effect can be influenced not only by the stage of 2HPT but also by the route of administration and quantity of calcitriol doses, as ensues from a long-term follow up of one patient. Moreover, the morphological substrate of the hyperplastic tissue of the parathyroid gland and their receptors for 1,25(OH)2D3 must be taken into account. Successfully performed parathyroidectomy, a still justified therapeutic step, is associated as a rule with rapid restoration of PTH levels. TO CONCLUDE: Pulse doses of calcitriol seem to be at present the effective treatment of diagnosed 2HPT, conventional oral calcitriol doses are useful in 2HPT prophylaxis. 2. The i.v. form should be the last resort of conservative treatment before parathyroidectomy. 3. Calcitriol treatment should attempt to maintain slightly raised PTH levels. 4. The limiting indicators of treatment are hypercalcaemia, hyperphosphataemia and the development of extraosseous calcifications. 5. In order to adhere to these criteria it is necessary to use dietary provisions, the dialyzation technique and check biochemical indicators of bone metabolism and possibly change doses of pharmaceutical preparations. PMID- 9750469 TI - [Hereditary hemorrhagic telangiectasis and the development of an arteriovenous fistula after 24 years]. AB - The authors describe a unique pulmangiographic picture taken 24 years after the first angiographic examination made on account of repeated haemoptysis associated with Rendu-Osler-Weber s disease. Angiography revealed enlargement of the aneurysm in the lungs and dilatation of the afferent and efferent vessel of the aneurysm. The only treatment of pulmonary arteriovenous fistulas is surgical. PMID- 9750468 TI - [Detection of myeloma cells in the peripheral blood using flow cytometry]. AB - Bone marrow plasma cells from patients with multiple myeloma express monoclonal cytoplasmic immunoglobulin, strongly express CD38 and usually coexpress CD56 and CD54. The aim of this study was to learn the relationship between the number of CD38+CD56+ and CD38+CD54 positive cells in peripheral blood with the activity of multiple myeloma. We evaluated the number of these cells in patients with monoclonal gammopathy and in the control group, as well. We used the two-color flow cytometry for this purpose. The peripheral blood of 57 patients with multiple myeloma and 4 patients with monoclonal gammopathy of unknown significance was repeatedly analyzed. Patients with high activity of multiple myeloma had the highest percentage of CD38+CD56+ and CD38+CD54+ cells. Patients with lower activity of the disease had lower count of these cells. But in 4 (11%) patients with high disease activity the count of CD38+CD56+ cells was zero. We conclude, that the longitudinal flow cytometric analysis of CD38+CD54+ and CD38+CD56+ cells is a useful method for estimation of disease activity in patients with multiple myeloma. Single analysis of these parameters cannot be used as a strict criterium for differential diagnosis. PMID- 9750470 TI - [Simultaneous occurrence of 2 thyroid gland carcinomas]. AB - The concurrent finding of two thyroid carcinomas is rare. The authors present two patients with the concurrent occurrence of two tumours. In the first case--a 51 year-old man--the authors found a medullary carcinoma in the right lobe and a medullary papillary carcinoma in the left thyroid lobe. The medullary papillary carcinoma metastatized into the mediastinal lymph nodes and into the mediastinal tissues in the area above the tracheal bifurcation. From the morphological and immunohistological aspect in the primary tumour of the left lobe as well as in the mediastinum two different clones of tumour cells were found (the medullary component of the tumour responded positively to antibodies against calcitonin and the papillary component to serum against thyroglobulin. The patient was treated by radiotherapy. The tumour progressed however markedly in its medullary component and the patient died one and a half year after the first symptoms from generalization of the tumour. The second patient, a woman, had an occult papillary carcinoma in the right lobe and in the left lobe a of a medullary papillary carcinoma (follicular variant). This type of tumour also metastatized into a cervical lymph node in the area of the left lobe. The patient was treated by radioiodine. Nine months after bilateral thyroidectomy she has no signs of progression of the tumour. PMID- 9750472 TI - [Thyrotropin-secreting adenomas of the hypophysis]. AB - Thyrotropin secreting pituitary adenomas are scarce. They may cause an extremely rare form of hyperthyroidism. The diagnosis is often delayed because the clinical symptoms are attributed to common types of hyperthyroidism. The diagnosis involves detection of elevated or normal (unsuppressed) thyrotropin levels in hyperthyroid patients and evidence of a pituitary adenoma by computed tomography or magnetic resonance imaging. The thyrotropin response in the thyrotropin releasing hormone test is either absent or insufficient. When the pituitary microadenoma appears to be undetectable, the familiar syndrome of selective pituitary resistance to thyroid hormones has to be excluded. Treatment involves extirpation of the tumour. If the macroadenoma is not removed completely, external radiotherapy of the pituitary follows. If this conventional treatment does not produce an adequate effect, treatment with long-acting somatostatin analogues is recommended. PMID- 9750471 TI - [Octreotide in the treatment of thyrotropin-secreting pituitary adenomas]. AB - The authors detected in a 30-year-old patient a very rare type of hyperthyroidism caused by a thyrotropin secreting pituitary adenoma. Scintigraphic examination of the pituitary by means of 111In radiolabelled octreotide revealed an increased accumulation of the radiopharmaceutical preparation in the tumour, which confirmed the high density of somatostatin receptors. After onset of octreotide treatment (Sandostatin, Sandoz, Switzerland) 3 x 100 ug/day by the s.c. route a brisk decline and normalization of thyrotropin already after the first dose was recorded. The thyroxine concentration declined slowly to the upper range of normal values. After 5 months treatment despite the positive response to receptor scintigraphy diminution of the adenoma was not recorded. Again an increase of thyrotropin above the upper limit of the reference range and a marked rise of thyroxinaemia were observed. Six months after radical selective trans-sphenoidal adenomectomy normal pituitary function was confirmed. PMID- 9750473 TI - [Relation between parathormone and 1,25-dihydroxyvitamin D3 in chronic kidney failure]. AB - The author gives an account of the basic relations of calcium homeostasis ensuing from the action of calcitropic hormones under physiological conditions and during chronic renal failure. The following are evaluated in relation to the pathogenesis of secondary hyperparathyroidism: a decline of glomerular filtration, the relationship of serum calcium levels and PTH secretion along with analyses of so-called S curves, calcitriol deficit and impaired function of the appropriate receptors, phosphate retention, effect of metabolic acidosis, participation of retained aluminium, possible importance of calcitonin and genetic abnormality, corticoid administration and abolition of oestrogen function. PMID- 9750474 TI - [Recommendations for the diagnosis and therapy of hyperlipoproteinemias in adults prepared by the Czech Atherosclerosis Society]. PMID- 9750475 TI - [Ultrasonographic diagnosis of parathyroid glands]. AB - Enlarged parathyroid glands are, if their localization is typical behind the thyroid gland, during basic USG examination with 5 MHz to 10 MHz using a linear probe, hypoechogenic or anechogenic, larger than 0.5 cm, oval in shape, elongated in a cranoiocaudal direction, separated from the thyroid gland. During swallowing they move upwards. Such a picture may be simulated also by enlarged lymph nodes, sympathetic ganglia a cranked vessel or oesophagus. By using coloured Doppler sonography in the algorithm of examinations the percentage of falsely positive findings may decline. Between 90 degrees and 270 degrees the vascular arch is sought which is formed by the branch of the a. thyroidea inferior. This vascular arch is found only when the parathyroid is involved, even when it is located in the parenchyma of the thyroid gland. PMID- 9750476 TI - [Lipoprotein(a)--a risk factor for coronary sclerosis in patients with hypothyroidism]. AB - In a group of patients with developed primary hypothyroidism the authors investigated in a longitudinal eight-month trial the effect of hormonal substitution therapy with thyroxine (T4) on the serum concentration of lipids, apolipoprotein B and lipoprotein (a)--risk factors for the development of early coronary sclerosis. In some patients--"responders"--gradually euhormonosis, normolipaemia are induced and clinical symptoms of hypothyroidism receded. In the second group of patients with hypothyroidism, so-called "non-responders" (n = 5) after eight months substitution treatment with thyroxine the anticipated effect does not occur and the investigated serum parameters improve only partially. The thyroxine, TSH levels and those of lipid parameters and apolipoprotein B persist in the zone of pathological values. The lipoprotein (a) concentration in both groups of patients with hypothyroidism does not change during thyroxine substitution and varies near baseline values. From the submitted observations the authors of the present work do not assume that thyroxine plays a part in the catabolism of lipoprotein (a) via LDL receptors, the activity and number of which increases along with the effect of thyroxine. PMID- 9750477 TI - [Fluvastatin in patients after heart transplantation]. AB - Hyperlipoproteinaemia is one of the frequent posttransplantation problems. Administration of statins is complicated in patients after transplantation by concurrent imunosuppressive treatment, in particular by possible undesirable interaction with cyclosporin. In the presented study 15 patients after transplantation of the heart with hyperlipoproteinaemia were examined who were on a standard triple combination of immunosuppressive drugs. Fluvastatin was administered, 20 mg in the evening, and in intervals of 6 weeks, 3 months and 6 months after the onset of treatment the levels of cholesterol, LDL and HDL cholesterol, triglycerides, urea, creatinine, liver terts and cyclosporine were followed up. The mean cholesterol level declined from 7.66 mmol/l during the 6rd week (p < 0.002), to 6.01 mmol/l during 3rd month and to 5.83 mol/l after the 6rd month (p < 0.001), LDL-cholesterol declined from 4.82 mmol/l and then 3.46 mmol/l and 3.31 mmol/l (p < 0.001). In the other investigated parameters no change recorded, incl. the cyclosporin levels. No clinical signs of muscular damage were recorded Fluvastatin thus does not only reduce effectively the cholesterol and LDL-cholesterol level but is also safe combination with immunosuppressive treatment. PMID- 9750479 TI - [Fatty acid and cholesterol crystals in bone marrow smears of oncology patients]. AB - In 50 haematological-oncological and oncological patients the authors found in bone marrow smears fatty acid and liquid cholesterol crystals. The number of crystals of fatty acids does not correlate directly with the triacylglycerol serum levels. Similar findings were recorded also in haematological patients. PMID- 9750478 TI - [Glutamic acid decarboxylase autoantibodies (antiGAD-Ab) in patients with non insulin dependent diabetes mellitus (NIDDM)]. AB - AIM OF STUDY: To assess the prevalence of markers of autoimmune destruction of pancreatic beta-cells in patients with non-insulin dependent diabetes mellitus (NIDDM). SUBJECTS: 127 hospitalized NIDDM patients subdivided to the following subgroups: non-obese with C-peptide < 0.3 nmol/l (NIDDM-(-)), non-obese with C peptide > 0.3 nmol/l (NIDDM-(+)), obese with C-peptide < 0.3 nmol/l (NIDDM+(-)) and obese with C-peptide > 0.3 nmol/l (NIDDM2+). METHODS AND MEASURED PARAMETERS: Age, BMI, C-peptide, autoantibodies to glutamic acid decarboxylase (antiGAD-Ab), autoantibodies to islet cells (ICA), markers of specific cellular immunity CD4, CD8, CD19, CD4/CD8, CD4/CD45/RA+, CD4/CD45/RA-, NK (CD16+56), CD3/HLADR, organ specific/non-specific autoantibodies. RESULTS: AntiGAD-Ab were positive in 5/15 (33.3%) NIDDM-(-), 1/32 (3.1%) NIDDM-(+), 2/9 (22.2%) NIDDM+(-) and in 3/71 (4.2%) NIDDM2+. The positivity of antiGAD-Ab in NIDDM-(-) and NIDDM+(-) was significantly higher (p < 0.05) than in NIDDM-(+) and NIDDM2+. CONCLUSION: Some patients with manifestation of diabetes in older age initially classified and treated as having NIDDM may have in fact slowly evolving autoimmune insulin dependent diabetes mellitus (LADA). These patients can be identified by measurement of antiGAD-Ab or other markers (ICA, IA-2) of autoimmune destruction of pancreatic beta-cells (AID). Moreover, in some patients both AID and insulin resistance may coexist in parallel. PMID- 9750480 TI - [Recurrent spontaneous dislocation of the catheter in a venous vascular access port system]. AB - The authors describe the problem of spontaneous changes of the position of the catheter in venous vascuports with a cannula inserted via the vena subclavia or internal jugular vein into the vena cava superior, used most frequently for repeated administration of chemotherapy. The correct position of the catheter is important in the venous port system for the proper function of the whole system. During dislocation of the catheter the function is impaired to a varying extent and the number of inflammatory and thrombotic complications increases and dislocations beyond the venous system lead to extravasal administration of the drug. Possible diagnosis of this condition and subsequent correction of the position of the cannula is described in a 48-year old man who developed spontaneous dislocation of the position of the cannula from the vena cava superior into the internal jugular vein. Correction by means of a hook inserted via the ipsilateral femoral vein into the vena cava inferior and via the vena cava superior into the internal jugular vein was successful only temporarily. Because of repeated dislocation the port system was removed. PMID- 9750481 TI - [Pitfalls in treatment with oral antidiabetic agents]. AB - In the treatment of type 2 diabetes (NIDDM) we possess three groups of oral hypoglycaemic drugs: sulfonyl urea derivatives, biguanides (metformin) and alpha glucosidase (acarbose) inhibitors. Oral treatment of diabetes has a favourable impact on the patients metabolic deviations but it involves also certain dangers and pitfalls. The side-effects of oral antidiabetics can be reduced to a minimum by respecting consequentially contraindications of administration of different preparations, knowledge of their mechanism of action and individual selection of a suitable antidiabetic for every patient. PMID- 9750482 TI - [Intracoronary stents today]. AB - The authors present an integrated view on the role of stents in different indications in patients with IHD from the aspect of recently published or presented randomized trials. PMID- 9750483 TI - [Endothelial cells and leukocytes--significance of adhesion interactions]. AB - Interactions between endothelial cells, adherence of endothelial cells to the molecules of extracellular matrix, adhesion of leukocytes and platelets to the activated endothelial cell lining are mediated by adhesion molecules belonging into four families: immunoglobulins, integrins, selectins, and cadherins. Adhesion interactions mediate intercellular signaling. Soluble forms of adhesion molecules modulate adhesion interactions. The determination of soluble adhesion molecules in blood is used for indirect evaluation of function or dysfunction of endothelial cells. It is possible to modulate abnormal function of endothelial cells using monoclonal antibodies against adhesion molecules. PMID- 9750484 TI - [Cardiology in the United Kingdom]. AB - The authors describe the contemporary postgraduate system of cardiology in Great Britain with regard to the unification of criteria for specialization in the European Union and discuss opportunities of doctors from the Czech Republic to join this postgraduate system. PMID- 9750485 TI - [Left ventricular remodeling in arterial hypertension and its therapy]. AB - Left ventricular (LV) hypertensive remodeling has polymorphic character with broad spectrum of its geometry. Echocardiographically it is simply possible to distinguish 4 LV remodeling types: normal geometry, concentric remodeling, eccentric hypertrophy and concentric hypertrophy. In an overview article morphological, functional and prognostic characteristics of remodeled hypertensive LV are presented. Most adverse prognosis as regards cardiovascular (CV) morbidity and mortality is in LV concentric hypertrophy. As to the current knowledge LV hypertrophy regression can be considered for potential aim of the management of arterial hypertension, independent from blood pressure normalization. Removal of structural changes of the CV system due to arterial hypertension could be more important function of antihypertensive drugs than blood pressure reduction alone. Until now minimally 147 articles regarding the influence of antihypertensive treatment on LV hypertrophy regression have been published. According to them most effective drugs are angiotensin-converting enzyme inhibitors. A new idea has brought the HYCAR study, which has demonstrated, that low dose of ramipril without any antihypertensive effect significantly reduces LV hypertrophy. The results of the study "evoke" change in the management of arterial hypertension: the treatment with low dose of ramipril in indication of LV hypertrophy regression also in those hypertensive patients with formerly normalized blood pressure. The importance of LV hypertrophy regression for the prognosis must be however specified in large designed trials, because until now it is not exactly known. PMID- 9750486 TI - [Development and progression of prostate cancer in relation with androgen dependency]. AB - Although carcinogen to human prostate cancer has not clarified yet, androgen is necessary for genetic damage on epithelial tissue of the prostate. There are many factors influencing promotion of cancer cells under certain androgenic environment; IGF, somatostatin, nutriments including beta-carotin, fat and green tea. Loss of androgen dependency gradually occur during progression. This may be attributable to adaptation, paracrine regulation and genetic change, but the latter is main course to acquire autonomous growth with metastatic ability. Mutation of androgen receptor which occurs in some cases plays a role on antiandrogen withdrawal syndrome. PMID- 9750487 TI - [Histological classification of prostatic cancer]. AB - Histological classification of prostatic cancer with a special focus on adenocarcinoma was reviewed according to "General Rule for Clinical and Pathological Studies on Prostatic Cancer (The 2nd Edition, 1992) published by Japanese Urological Association and The Japanese Society of Pathology. The points of the classification are as follows; (1) adenocarcinoma is separated into 3 categories, namely, well, moderately and poorly differentiated types, by structural features. (2) nuclear grading does not commit for making a subclassification of prostatic adenocarcinoma. The other types of primary malignancies are rare in the prostate. Prostatic intraepithelial neoplasia should be discussed in the further revision of the classification. PMID- 9750488 TI - [Clinical staging of prostate cancer]. AB - Clinical staging of prostate cancer was reviewed on the basis of TNM classification edited by UICC in 1997. In this revision, (1) T1c was newly categorized among T1 for cases of high PSA level without any abnormal sign of DRE, (2) T2 was subdivided into T2a and T2b in accordance with unilateral or bilateral nodule in the prostate, respectively, (3) T3 was also subdivided into T3a with capsular invasion and T3b with seminal vesicle invasion. It is hoped that present classification will become useful tools for the detection of the tumor burden cancer patients. PMID- 9750489 TI - [Epidemiology and clinical features of prostate cancer in Japan]. AB - The incidence rate of clinically manifest prostate cancer in 1992 was estimated 15.7 per 100,000 men, although it is increasing exponentially. Accordingly, 5399 deaths from prostate cancer in 1995 will be increased to 13,494 deaths in 2015. Change in dietary habit (more Western-style diet) is considered to be a major cause of the increase. Escalating number of elderly people in the Japanese population is another major reason of elevated incidence. On the other, public awareness of prostate cancer and introduction of serum PSA measurement to health check-up undoubtedly have raised the detection rate of early stage disease. The way of androgen ablation do not seem to have influenced on survival of the advanced disease so far. It remains to be clarified whether the combined androgen blockade using pure anti-androgens with castration provide better patients' survival than castration alone. PMID- 9750490 TI - [Prostatic mass screening program and its technology]. AB - In 1975, we originally developed a mass screening program for prostatic cancer (PC) using transrectal sonography (TRS). In the primary study, males more than the age of 55 years were examined by TRS and subjects with any pathological findings on the sonogram were referred to secondary examination. In the secondary examination, biopsy of prostate was carried out under ultrasonic guidance. Since 1984, we included digital rectal examination (DRE) in the primary study. We further studied prostate specific antigen (PSA) in 1987. From 1995, mass screening program for PC using PSA has been performed. In the primary study, subjects were examined by PSA and the cut-off values were set at 4 and 10 ng/ml. Subjects whose PSA level ranged from 4.1 to 10.0 ng/ml (gray zone) were examined by DRE, TRS and PSA density (more than 0.15) in the secondary study, then the suspected subjects of PC underwent 6 sextant biopsies under interventional ultrasound. Subjects whose PSA level was more than 10.1 ng/ml were biopsied directly. According to a survey by the Foundation for prostate Research, the similar field screening has been performed in Japan by 32 groups for a total of 97,066 subjects, detecting 817 (0.8%) cancer cases up to 1995 and 431 of them (53%) were in the early stage. Prostatic screening is thus being recognized as an important part of preventive oncology at a national level in Japan. PMID- 9750491 TI - [Evolution of treatments for prostatic cancer]. PMID- 9750492 TI - [Clinical staging and management of prostate cancer]. AB - Through the use of digital rectal examination, prostate-specific antigen and improved biopsy technique, it is possible to diagnose early stage prostate cancer. The management of this disease, however, has generated considerable controversy. The decision whether to treat actively or conservatively must be made on the basis of accurate staging and grading technique, a patient's life expectancy and performance status. This article will show the current staging and management of prostate cancer as practiced in Japan. PMID- 9750493 TI - [The usefulness of prostate-specific antigen assay for the early detection and follow up of the prostate cancer]. AB - PSA has been widely used as a tumor marker for screening, diagnosis and monitoring of prostate cancer. We demonstrated the 3 of 7 (43%) patients with a serum PSA level of < or = 4 ng/ml, 12 of 24 (50%) patients with a serum PSA level ranging from 4.1 to 10.0 ng/ml, 10 of 17 (59%) patients with a serum PSA level ranging from 10.1 to 20 ng/ml, 6 of 8 (75%) patients with a serum PSA level ranging from 20.1 to 30.0 ng/ml, 5 of 6 (83%) patients with a serum PSA level ranging from 30.1 to 40.0 ng/ml, 29 to 29 (100%) patients with serum PSA level of > or = 40.1 ng/ml had prostate cancer. It is recommended that the use of PSA and DRE in combination is important as a diagnostic procedure for the early detection of prostate cancer. PMID- 9750494 TI - [The significance and limitation of prostate specific antigen in the mass screening for prostate cancer]. AB - The ultimate aim of the mass screening program is to decrease the prostate cancer mortality, which can be demonstrated only in a well designed and well controlled trial comparing screened and unscreened populations. However, there are not any trials which prove the effectiveness of mass screening for prostate cancer. Among the 3 modalities, prostate specific antigen (PSA) had the highest sensitivity and positive predictive value. However, PSA had the 20% probability to diagnose incorrectly the patient with prostate cancer as a normal. PSA had 10% possibility to detect clinically insignificant cancer. It is important to inform subjects not only the significance but also limitation of mass screening by PSA. PMID- 9750495 TI - [Standardization of PSA assays]. AB - Presence of various PSA assays and no uniform serum data have led the clinicians confuse at the evaluation of serum data. Therefore, the present status of standardization of prostate-specific antigen (PSA) assays and some attempts for obtaining the same serum data were presented. With the combination of so called Stanford reference for calibration standard and the linear regression line proposed by Japanese Urological Association, the almost same serum PSA values might be obtained. However, the characteristics of each PSA assay such as equimolar or skewed type must be taken into consideration at the use. PMID- 9750496 TI - [Clinical significance of prostate specific antigen and gamma-seminoprotein ratio for diagnosing prostate cancer]. AB - It has been reported that prostate specific antigen and gamma-seminoprotein ratio (PSA/gamma-Sm ratio) is an useful means for distinguishing benign prostatic hyperplasia and prostate cancer if serum PSA is measured by Eiken-PSA method. We studied the clinical significance of PSA/gamma-Sm ratio when using Markit-M-PSA method. PSA/gamma-Sm ratio had no superiority over PSA alone for detecting prostate cancer. The present results suggest that the clinical significance of PSA/gamma-Sm ratio can be varied by various PSA-assay kits. PMID- 9750497 TI - [Percent free prostate-specific antigen: a marker for detection of prostate cancer]. AB - We reviewed the use of percent free prostate specific antigen(PSA) to aid in the differentiation of benign and malignant prostate diseases. Percent free PSA may increase the sensitivity and specificity of PSA testing. There is evidence to suggest a benefit cost advantage to a tailored biopsy approach based on percent free PSA. However, differences in study designs and subject populations may account for the confusion in the current literature. Therefore, statistically valid multicenter clinical trials that take into account influencing factors are needed to set assay specific cut points and probability determinations. PMID- 9750498 TI - [The efficacy of PSA density for the early detection of prostate cancer]. AB - OBJECTIVE: We studied on the efficacy of prostate specific antigen density (PSAD) for the detection of prostate cancer among patients with intermediate serum PSA levels. MATERIALS AND METHODS: Transrectal ultrasonography (TRUS) and transrectal prostate biopsy were performed in 103 patients whose PSA levels were 10 ng/ml or less despite positive digital rectal examination(DRE) or whose PSA levels were intermediate (4 to 10 ng/ml). Prostate volume was determined by TRUS and PSAD was calculated (serum PSA divided by volume of entire prostate volume). The rate of positive biopsy was compared with PSAD (more than 0.15 versus less than 0.15), DRE (positive versus negative) and patient's age (more than 61 versus 60 or less). RESULTS: The overall cancer detection rate was 43.7% in this study. There was no apparent correlation between patient's age and cancer detection rate when the patient's age was more than 61. DRE itself was not effective for the detection of prostate cancer in the patients whose PSA level was 10 ng/ml or less. Independent of DRE findings, the rate of positive biopsy was double in the patients whose PSAD was more than 0.15, compared with the patients whose PSAD was less than 0.15. CONCLUSIONS: For the early detection of prostate cancer, PSA density may be useful in the selection of patients for transrectal prostate biopsy. PMID- 9750499 TI - [PSATZ (prostate specific antigen adjusted for the transition zone volume)]. AB - This brief review discusses the diagnostic ability of prostate specific antigen adjusted for the transition zone volume (PSATZ) for the detection of prostate cancer in patients with intermediate prostate specific antigen (PSA) levels. PSATZ was defined as the quotient of the PSA value and the calculated transition zone volume. In comparison with standard parameters including PSA and prostate specific antigen density, PSATZ could be a useful indicator for the detection of prostate cancer in patients with PSA values between 4.1 and 10.0 ng/ml, especially in those with normal digital rectal examinations. Similar observations consistent with our results have been also reported by other investigators. PSATZ has some disadvantages including volumetry and expensive cost. Further study is necessary to discuss whether PSATZ is superior to other new modalities such as free-to-total PSA ratio with regard to diagnostic cost and efficacy. PMID- 9750500 TI - [The clinical value of prostate-specific antigen velocity as a method for prostate cancer detection]. AB - The usefulness and problems associated with measuring of prostate-specific antigen (PSA) velocity (PSAV) for detecting prostate cancer are reviewed. PSA is not a cancer-specific serum marker, and various physiologic and benign pathologic processes influence serum PSA concentrations. Thus, it is important to distinguish between the elevation of serum PSA caused by cancerous tissue and biological variations. Smith suggested that a PSAV cutoff point of 0.75 and 0.4 ng/ml/year or more maximized the sensitivity and specificity of predicting cancer in those with normal PSA levels and elevated PSA levels (greater than 4.0 ng/ml), respectively. We also demonstrated that PSAV is significantly higher in moderately to poorly differentiated cancer than that in well differentiated cancer. PMID- 9750501 TI - [Prostate specific antigen doubling time in prostate cancer before treatment and in refractory status]. AB - Prostate-specific antigen(PSA) increases exponentially in prostate cancer patients before treatment and in refractory status. PSA increases in 68-86% of prostate cancer patients before treatment, and that of the remaining 14-32% of the patients is stable. Those patients with a higher pre-treatment PSA level are more likely to have a shorter PSA-doubling time(PSA-DT). The relationship between pre-treatment stage, grade and PSA-DT is controversial. PSA-DT in biochemical failure patients predicts the risk of clinical recurrence. PSA-DT was correlated well with time to clinical recurrence after biochemical failure. Distant recurrence was associated with short PSA-DT. Higher clinical stage and lower differentiation before treatment correlated with shorter PSA-DT in recurrent cancer patients. PSA-DT is an important parameter for judging malignant potential of each cancer. PMID- 9750502 TI - [Progress of clinical application for prostatic ultrasound]. AB - In 1967, in our laboratory the first clinically available prostatic sonogram was obtained from the rectal cavity with a patient in the lithotomy position. Thereafter, transrectal sonography (TRS) visualized the internal architecture as well as the entire contour of the prostate, and was readily applied to the investigation of prostatic diseases. Currently, TRS occupies a central position among the diagnostic modalities for the prostate. Along with accumulating evidences indicating the clinical usefulness of TRS in the diagnosis of prostatic diseases, recent development has confirmed it to be indispensable for daily practice in urology clinics. This chapter will describe an overall review of the development and diagnostic criteria of TRS. PMID- 9750503 TI - [Efficacy of transrectal ultrasonography in staging prostate cancer]. AB - Clinical staging can be defined as the attempt to determine the pathologic extent of cancer by clinical tests. Accurate preoperative assessment is essential for the appropriate selection of therapy and design of treatment for the individual patients with prostate cancer. Transrectal ultrasonography (TRUS) was initially applied clinically in 1971. It is currently considered an essential tool in the management patients with prostatic disease. More importantly, ultrasound guidance allows accurate sampling of prostatic tissue. However, the role of sonographic imaging itself in diagnosing of tumor extent remains uncertain. In this chapter we will assess critically the present role of ultrasound in staging prostate cancer based on currently available criteria. PMID- 9750504 TI - [Clinical role of magnetic resonance imaging (MRI) of prostatic cancer]. AB - Magnetic resonance imaging (MRI) has become very important tool for diagnosis in many fields including neurology and orthopedics. In this article, the usefulness and current status of MRI in the clinical diagnosis of prostate cancer are discussed. The advantage of MRI over the other methods like ultrasound imaging and CT scan is that zonal anatomy of the prostate gland is clearly identified into peripheral, central and transition zone. Cancer in the peripheral zone is demonstrated as a low intensity lesion in T2-weighted images but cancer in other zones are difficult to be identified. Extracapsular invasion, seminal vesicle invasion and neuro-vascular invasion can be identified which is useful for the assessment of tumor extension. MRI is also useful for the detection of bone metastasis. PMID- 9750505 TI - [Assessment of local staging of prostate cancer by endorectal surface coil]. AB - We assessed the usefulness of endorectal surface coil MRI (ERSC-MRI) for staging diagnosis of prostate cancer to compare preoperative ERSC-MRI findings with pathological staging in patients performed radical prostatectomy. MR imaging was performed on a 1.5 T MR system with an endorectal surface coil designed for imaging the prostate. At the time the coil was inserted, 1.0 mg of glucagon was injected in tramuscularly. T1-weighted MR images were obtained in the axial plane, and T2-weighted, spin echo MR images were obtained in the sagittal, coronal, and axial planes for each patient. The capsular penetration of prostatic cancer was defined according to the six diagnostic criteria by Outwater et al.: (1) a bulge formation of the low-signal-intensity area beyond the prostatic capsule, (2) low-signal-intensity stranding in the periprostatic tissue, (3) retraction of the prostatic capsule besides the low-signal-intensity area, (4) elongation of the low-signal-intensity area in the prostatic capsule, (5) thickening of prostatic capsule. (6) extracapsular tumor. The sensitivity, specificity and accuracy of ERSC-MRI for capsular penetration of the prostatic cancer were 95.5%, 40.0%, and 85.2%, respectively. These results indicate that ERSC-MRI is useful for staging diagnosis of prostatic cancer. PMID- 9750506 TI - [Fluorodeoxyglucose positron emission tomography in diagnosis of untreated prostate cancer]. AB - Fluorodeoxyglucose positron emission tomography (FDG-PET) has been utilized since glucose metabolism in cancer tissue is considered to relate to its malignant potential. We evaluate the application of FDG-PET to prostate cancer. FDG uptake of cancer tissues was higher with higher Gleason grade, advanced clinical stage and higher serum PSA value than that with lower Gleason grade, localized clinical stage and lower serum PSA value. The measurement of glucose metabolism in the prostate using FDG-PET was suggested to be useful for evaluation of the biological malignant potential of prostate cancer. It may serve as a predictive factor for prognosis and may be useful in determining the effectiveness of hormonal therapy in prostate cancer. PMID- 9750507 TI - [The advance of ultrasound guided prostate biopsy--comparison between 4 quadrant and 6 sextant biopsy]. AB - Although systematic biopsy has increased the detection rate of prostate cancer, the optimal method of biopsy has not yet been fully established. The number and site of cores, and the biopsy route are controversial in terms of cancer detection and complication. We briefly review the advances in prostate biopsy, and present the results of our biopsy methods. Our study showed that the difference of cancer detection rate between 4 quadrant and 6 sextant biopsy was not significant. There was little value in systematic transition zone biopsies. However, such biopsies proved useful in patients whose first systematic biopsies was negative and who have persistently elevated PSA values. It is recommended that the biopsy protocol for routine prostate cancer detection be targeted to the peripheral zone. PMID- 9750508 TI - [Diagnosis of organ-confined prostate cancer by systematic biopsy under guidance of TRUS]. AB - Systematic biopsy is useful to diagnose of prostate cancer. However, the usefulness of systematic biopsy for detection of organ-confined prostate cancer is unclear. There are three biopsy parameters, 1) percentage of biopsy cores involved with cancer (number of cores involves with cancer divided by total number of biopsy cores), 2) millimeters of cancer per biopsy core (total millimeters of cancer in the biopsy specimen divided by total number of biopsy cores) and 3) percentage of cancer in the biopsy specimen (total millimeters of cancer in the biopsy specimen divided by total millimeters of biopsy tissue). Although each biopsy parameter alone is not sufficient to detect organ-confined prostate cancer, the combination with these biopsy parameters and serum prostate specific antigen levels improved the detection rates of organ-confined prostate cancer. PMID- 9750509 TI - [Transrectal ultrasound guided transperineal systematic prostate biopsy]. AB - We retrospectively reviewed the files of patients who had transperineally undergone a systematic six sextant prostate biopsy under guidance of transrectal ultrasound in longitudinal transsection by using Biopsy needle with the spring roaded Biopsy gun. Of the 442 patients with suspicious glands visiting our clinic between May in 1992 and February in 1998, 486 biopsies were taken and pathological diagnosis was made to be prostate cancer in 153 (34.6%) patients, benign prostate hyperplasia in 270 and prostatitis in 19. Positive predictive values obtained in this series were calculated to be 49.4% in digital rectal examination (DRE), 55.0% in transrectal ultrasonogram (TRUS), 47.8% in prostate specific antigen (PSA) level 4 ng/mL or greater and 83.5% if a patient had positive DRE and TRUS and PSA level 4 ng/mL or more. Relationship between number of positive cores and clinical stage of the patients was found (p < 0.0001). No complications relating to the biopsy were observed. This transrectal ultrasound guided transperineal systematic six sextant prostate biopsy was thought to be one of useful tools of the biopsy for detecting prostate cancer. PMID- 9750511 TI - [The clinical usefulness of urinary pyridinoline and deoxypyridinoline as potential markers of bone metastasis in patients with prostate cancer]. AB - The levels of probable markers of bony metastatic disease were measured to evaluate their efficacy as predictors of disease and therapeutic outcome. Urinary pyridinoline and deoxypyridinoline were measured in patients with benign prostatic hyperplasia, clinically localized prostate cancer and prostate cancer with bone metastases. Also, urinary pyridinoline and deoxypyridinoline were compared in 2 groups of patients with metastatic prostate cancer of the bone who demonstrated progression or positive response to treatment. Urinary pyridinoline and deoxypyridinoline were determined by high performance liquid chromatography and were normalized to urinary creatinine. Levels of urinary pyridinoline and deoxypyridinoline in urine in patients with bony metastatic prostate cancer were significantly greater than levels in patients with benign prostatic hyperplasia or localized prostate cancer. Serial measurement of urinary pyridinoline and deoxypyridinoline was correlated with a positive response to treatment (decreased) and with clinical progression of disease (increased) before detection of new bone lesions by bone scintigraphy. Measurement of urinary pyridinoline and deoxypyridinoline may provide a useful marker of prostate cancer metastatic to bone and may be useful in monitoring the response to treatment. PMID- 9750510 TI - [Clinical usefulness of blood PICP, PINP and ICTP concentrations as bone metastasis markers in prostate cancer patients]. AB - PICP and PINP are considered markers of bone formation, and ICTP is considered to be a marker of bone resorption. We measured these three markers as indicators of bone metastasis of prostatic cancer, and assessed their clinical usefulness. Serum PICP, PINP and ICTP were significant higher in patients with bone metastasis than localized cancer or controlled bone metastasis. Serum PSA increased initially with the LH-RH agonist during the course of bone metastasis. All these markers were subsequently observed to increase. Change in PICP of patients with bone metastasis was slight compared to PINP during endocrine treatment. Serum PICP, PINP and ICTP reflect the extent of metastasis in bone and are useful for monitoring the response of this condition to therapy. PMID- 9750512 TI - [Application of radical prostatectomy for prostate cancer and its outcome]. AB - The proportion of patients with localized prostate cancer treated by radical prostatectomy is increasing rapidly in Japan. As for the qualifications of patient candidates for radical surgery, various clinical and pathological findings to predict tumor extent and disease-free outcome must be considered carefully. There has been increased interest in the application of neoadjuvant or adjuvant therapy for locally advanced tumor group in order to improve disease free survival and overall survival. The new anatomical approach to radical prostatectomy with its nerve sparing option assures preservation of erection. This procedure achieves excellent cancer control for patients with a definite organ-confined tumor preoperatively. Finally, more time is needed to obtain information on the long-term outcome after radical prostatectomy. PMID- 9750513 TI - [Decision making analysis in the treatment of prostate cancer]. AB - The decision analysis technique has been used to estimate the benefits of treatment strategies for a variety of diseases. This analysis demonstrated that a selective treatment with a reasonable specificity and sensitivity for detecting clinically significant cancer is more beneficial to the patients with T1c cancer under 70 years of age than radical prostatectomy or watchful waiting assigned to all patients. Moreover, it was clearly indicated that quality-of-life (QOL) is the most important factor for deciding the optimal treatments of prostate cancer. PMID- 9750514 TI - [The definition and frequency of clinically unimportant prostate cancer in the patients treated with radical prostatectomy]. AB - In autopsy studies more than 20% of men > 50 years old have prostate cancer, yet the lifetime risk of developing clinical cancer of the prostate is much less. Thus, early detection should not aim to all cancers, but only those that are potentially morbid or lethal. In the detailed morphometric examination of radical prostatectomy specimens, we found only 10% of the 610 patients had clinically unimportant cancer. Of the 207 patients with non-palpable tumor, 11% had clinically unimportant cancer compared to 10% of the 403 patients with palpable tumor (p = 0.52). And these non-palpable tumor were more likely to be curable disease compared to palpable tumor (66% versus 57%, p = 0.007). All recent studies indicated that most of prostate cancers which were detected with current diagnostic tests were clinically important and, therefore, should be treated aggressively. PMID- 9750515 TI - [Quality of life after radical prostatectomy]. AB - In the past, radical prostatectomy commonly led to urinary incontinence and erectile dysfunction. In the last decade, new operative techniques have greatly reduced the complication rate and the operation has gained increasing popularity as treatment of choice for localized prostate cancer. Success or failure of radical prostatectomy has been reported not only in terms of disease-free survival, but in terms of patient attitudes to treatment and side effects. As physicians, we must remember that in presenting treatment options to patients it is important to emphasize both the quality and quantity of life that may result. With richer information on QOL in addition to duration of survival, patients will be able to make more informed decisions. Therefore, the QOL study will contribute patient self-report data to current treatment decision models that rely solely on physician estimates of patients' QOL and side effects following radical prostatectomy. We herein report the results of our recent QOL survey in men treated with radical prostatectomy, and briefly discuss QOL methodology. PMID- 9750516 TI - [Impact of life expectancy on the clinical significance and curability of prostate cancer]. AB - Theoretical projected prostate cancer volume at the time of expected death was determined based on patient age and index cancer volume at diagnosis, assumed cancer volume doubling time and life expectancy of Japanese male population. Clinically insignificant cancer in 104 prostatectomy specimens was found to occur at 4.8, 10.6, 15.4 and 26.9% for tumor doubling times of 2, 3, 4 and 6 years, respectively. Assuming a 2-year doubling time with clinically insignificant cancer excluded, only 36.4% of significant cancers could be considered potentially curable and with a 3-year doubling time, 32.3%. For 4- and 6-year doubling times, only 30.7% and 25.0% of the clinically significant cancers were potentially curable, respectively. Patient life expectancy and tumor doubling time significantly determine the outcome of treatment for prostate cancer especially in elderly males with higher risk of mortality. PMID- 9750518 TI - [Surgical or medical castration with LH-RH analogue for prostatic cancer]. AB - Since the discovery by Huggins and Hodges that androgen deprivation therapy is effective for prostatic cancer, surgical castration or the administration of exogenous estrogen has been the mainstay of treatment for advanced prostatic cancer. However, surgical castration is refused by some patients because of psychological impact and estrogen therapy is reported to be associated with significant morbidity and mortality. The chronic administration of superactive analogue of LHRH has been shown to suppress markedly gonadal steroidogenesis. Medical castration with LHRH analogue has been demonstrated to be safe and effective in patients with advanced prostatic cancer. But the traditional surgical castration still remains as another powerful option, because it is more excellent in the points of compliance and cost-benefit than a monthly injection of expensive LH-RH analogue. PMID- 9750517 TI - [Recent advance in luteinizing hormone-releasing hormone analogue therapy in management of prostate cancer]. AB - Luteinizing hormone-releasing hormone (LHRH) analogue therapy is one of the most widely used hormonal therapy for prostate cancer. LHRH analogue is effective and safety therapy for prostate cancer, when appropriate consideration is given to the disease flare. Recently, new long acting 3-month LHRH analogue depot has been developed and showed the therapeutic equivalence to the 1-month depot. In basic research, the expression of LHRH and its receptor has been demonstrated in some kinds of hormone depend cancer including prostate cancer. Although precise role of LHRH system in those cancers remains unclear, these findings suggests the presence of an autocrine system based on LHRH. More understanding of LHRH system might provide new treatment approaches to those cancer. PMID- 9750519 TI - [Assessment of the quality of life (QOL) of prostatic cancer patients under LH-RH analogue treatment]. AB - Assessments of QOL on 53 prostatic cancer patients under LH-RH analogue treatment were evaluated using our own questionnaire. Three months after treatment, the improvement of ADL, appetite, physical symptoms, mental symptoms, sexual life and social life occurred in 24.3%, 51.5%, 44.7%, 29.7%, 7.5% and 35.4% of the patients, respectively. Twelve months after treatment, the improvement of them occurred in 30.9%, 48.4%, 31.7%, 33.7%, 8.3% and 34.4%, respectively. The quality of sexual life for the prostatic cancer patients treated with LH-RH analogue was worse than that of others. LH-RH analogue is effective on QOL in the treatment of prostatic cancer patients. PMID- 9750520 TI - [Steroidal and nonsteroidal antiandrogens: chemical structures, mechanisms of action and clinical applications]. AB - Antiandrogens are defined as substances which prevent androgens from expressing their activity at target sites. Based on their chemical structures, antiandrogens are divided into steroidal and nonsteroidal antiandrogens. In addition to antiandrogenic action, steroidal antiandrogens simulate the negative feedback inhibition of the hypothalamus, resulting in lowering the concentration of plasma testosterone. On the other hand, in nonsteroidal antiandrogens, the androgenic actions are blocked in both hypothalamus and target tissues. Therefore, negative feedback signals are inhibited and production of testosterone is increased in the testis. For the endocrine therapy of prostate cancer, chlormadinone acetate, flutamide and bicalutamide are or will be available in Japan. Antiandrogen is generally used by the combination with surgical or medical castration, namely as total androgen blockade. PMID- 9750521 TI - [Total androgen blockade--concept, theory, method and clinical application]. AB - There can be seen many investigations to examine the effects and benefits of total androgen blockade (TAB), combining an antiandrogen with surgical castration or a LH-RH analogue, for advanced prostate cancer. This review summarizes the concept, theory, method and clinical application of TAB. The concept of TAB was supported by reports that show a survival advantage using the combined blockade over LH-RH analogue alone. The theory of TAB proposes that suppression of all androgen production, adrenal and testicular androgen, should result in a better response than standard hormonal management such as castration and/or estrogens. In Japan, Chlormadinone acetate (100 mg twice daily) or Flutamide (375 mg three times daily) is orally administered and Leuprorelin acetate (3.75 mg every 4 weeks) or Goserelin acetate (3.6 mg every 4 weeks) is administered by hypodermic injection. There have been unresolved controversies surrounding this therapeutic modality, therefore future studies should help to define the role of TAB. PMID- 9750522 TI - [Antiandrogen withdrawal syndrome]. AB - The antiandrogen withdrawal syndrome was first reported in patients with prostate cancer who manifested disease progression after total androgen blockage therapy with medical or surgical castration and pure antiandrogen, flutamide; discontinuation of flutamide resulted in a decline in prostate specific antigen and, in some cases, with clinical response. Same phenomena have been reported after the withdrawal of casodex, chlormadinone acetate, megestrol acetate, diethylstilbestrol, and estramustine phosphate. Mutations in the androgen receptor(AR) gene were discovered in clinical specimens of human prostate cancer patients who showed this syndrome and some of these being identical to the mutation found in LNCaP prostate cancer cell line. Another mechanism otherwise the point mutation of AR would be also speculated. PMID- 9750523 TI - [Reevaluation and potential therapeutic effects of estrogen therapy to prostate cancer patients]. AB - Survival time of 65 patients with stage D2 prostate cancer who were treated by estrogen with or without antiplatelet drugs was examined. Survival time of 37 patients with antiplatelet drugs was significantly longer than that of 28 histological control ones. The frequency of death of cardiovascular disease in the former group decreased to 5.3%. Next, progression free time of the 37 patients was compared to the time of 37 patients treated by antiandrogen therapy. The former one was better than the latter. However, survival time after relapse of the estrogen group was shorter than the control. These results suggest that estrogen therapy improved by a combination of antiplatelet drugs has to be reevaluated and contains specific therapeutic effects differing from antiandrogen therapy. PMID- 9750524 TI - [High-dose intravenous diethylstilbestrol diphosphate therapy for hormone refractory prostate cancer]. PMID- 9750525 TI - [Application and limitation of neoadjuvant hormonal therapy for prostate cancer]. AB - Of 156 patients, 111 (clinical stage T1a-b; 21, T1c; 17, T2a-b; 36, T2c; 27, T3; 10) immediately underwent radical prostatectomy (surgery group), and 45 (clinical stage T1a-b; 8, T1c; 4, T2a-b; 10, T2c; 9, T3; 14) received neoadjuvant hormonal therapy (NHT group). NHT offered probability of increasing organ-confined cancer(OCC; pathological stage pT2 or lower N0M0) in the following group, which contains (a) patients who had moderately differentiated adenocarcinoma in the biopsy specimen and T2b or lower diseases, and (b) those who had well differentiated adenocarcinoma, T2c diseases and PSA levels of 10 ng/ml or higher, referred to as "OCC suitable criteria". Of 156 patients, 51 (33%) met OCC suitable criteria. In those cases, the proportion of OCC in NHT group was significantly higher than that in surgery group (11/12 (92%) vs. 16/39 (41%), p = 0.002). NHT is useful for increasing OCC in patients who meet OCC suitable criteria. PMID- 9750526 TI - [A study on androgen deprivation therapy prior to radical prostatectomy with special reference to the tumor volume]. AB - BACKGROUND: The aim of this study is to analyze correlation between the tumor volume (TV) and the final pathological stage in patients undergoing neoadjuvant therapy. PATIENTS AND METHODS: Twenty-six patients underwent radical prostatectomy alone (group S). The remaining 28 patients had undergone neoadjuvant therapy (group N) which mainly consisted of LH-RH agonist. TV and the final pathological stage were retrospectively reviewed. RESULTS: The likelihood of positive surgical margin was statistically higher in group S than in group N (p = 0.009). The correlation between TV and the pathological stage was observed not only in group S (rho = 0.646, p = 0.0012) but also in group N (rho = 0.716, p = 0.0002). The mean TV was statistically smaller in group N in patients with baseline PSA level more than 10.1 ng/ml (p = 0.024). CONCLUSION: A decrease in tumor volume caused by neoadjuvant therapy may play an important role in reducing the likelihood of positive surgical margin. PMID- 9750528 TI - [Chemotherapy for hormone-refractory prostate cancer]. AB - Chemotherapy may be an appropriate treatment for patients with evidence of disease progression despite of antiandrogen withdrawal or secondary hormonal therapy. But none of the chemotherapeutic regimens, neither monotherapy nor combination therapy, were shown to be superior to another with regard to survival. The criteria to assess the effect of chemotherapy on disease-related symptoms and quality of life is expected to be framed. Preclinical investigations and well-designed, well-powered clinical trials remain key to altering the natural history of hormone-refractory prostate cancer. PMID- 9750527 TI - [Neoadjuvant androgen deprivation preceding to radical prostatectomy--its role in short-term and long-term outcomes]. AB - Neoadjuvant androgen deprivation has been demonstrated to reduce the risk of surgical margin positivity and lymphnode metastasis, and facilitate a thorough prostatectomy. On the contrary, no long-term studies have proved the beneficial effects of preoperative therapy on patients' survival. A multicenter randomized study in Japan comparing neoadjuvant therapy and immediate surgery (control) was conducted. All the patients were to receive hormonal therapy for 2 years. The results confirmed the short-term efficacy of neoadjuvent hormonal treatment, and no significant difference in clinical relapse rate (5.6% (5/90) in neoadjuvant group versus 11.6% (10/86) in control group), nor PSA relapse rate (13.3% versus 17.4%) at 2 years posttreatment; however in stage C cancer there was a tendency of a lower clinical relapse rate in neoadjuvant group (9.5%: 2/21) than the control (30.4%: 7/23). These data provide no obvious evidence for favorable long term outcomes in neoadjuvant group, although a possibility that stage C cancer benefits from the treatment remains to be explored. PMID- 9750529 TI - [Usefulness of initial chemoendocrine therapy for advanced prostate cancer]. AB - The 5 year cancer specific survival rate of advanced prostate cancer, especially in metastatic cancer is less than 40%. Recently, maximum androgen blockade showed some beneficial effects in cases of minor disease but no additional usefulness in major cases. The treatment modality referred to as initial chemoendocrine, used to treat prostate cancer, seems to be a reasonable method because prostate cancer cells contain heterogeneity. This procedure means that the endocrine treatment is best suited to treat hormone sensitive cells, whereas chemotherapy is more appropriately used as a firstline therapy for hormone insensitive cells. We reported that the initial chemoendocrine method showed superiority in the 5 year cancer specific survival category than in the endocrine therapy analyzing non randomized trials. From that stage on we reviewed the beneficial point of the treatment, and are now trying randomized control studies. PMID- 9750530 TI - [Intraoperative radiotherapy combined with external beam radiation for prostate cancer without metastasis]. AB - Between 1989 and 1996, 35 patients with prostate cancer without metastasis received intraoperative radiotherapy combined with external beam radiation. 10 of 16 stage B patients and all of 19 stage C patients received additional endocrine therapy for the initial treatment. The radiation therapy included 25-30 Gy of intraoperative radiotherapy for prostate and 30 Gy of external beam radiotherapy for small pelvic region. One patient of stage C was dead for cancer and 4 patient were dead for other causes during 15-99 (mean: 41.6) months follow up period. The overall actuarial survival at 5 years by Kaplan-Meier method were 92.3% for stage B and 87.2% for stage C. Although cystitis, proctitis and anal bleeding were observed as the adverse effects of radiotherapy, both acute and chronic symptoms were not critical. In conclusion, intraoperative radiotherapy combined with external beam radiotherapy was revealed as an effective treatment for prostate cancer without metastasis. PMID- 9750531 TI - [Radiation therapy with neoadjuvant hormonal therapy for prostate cancer confined to pelvis]. AB - Neoadjuvant endocrine treatment prior to radical prostatectomy for prostate cancer confined to pelvis has of some value to prevent progression although there are many controversies. In order to improve the prognosis of locally advanced prostate cancer (stage B2 and C), definitive radiotherapy with neoadjuvant endocrine therapy has been investigated. Endocrine therapy reduces the volume of prostate, thus reduces the amount of side effect via reducing the area of irradiated normal tissue. Effect of radiation and that of endocrine therapy to induce apoptosis might be synergistic. The result is favorable although the follow-up period is too short. Further studies are needed to make conclusion. PMID- 9750532 TI - [Trend in genetic medicine]. AB - Genetic medicines including antigen, antisense molecules, ribozymes, aptamers, plasmid, gene have been enthusiastically studied as pharmaceutical agents. In the past decade, genetic medicines have been bogged down in solving serious problems, especially delivery efficiency. Steady study are continued to apply chemical modifications, cationic liposomes or another drug carriers enhance biological activities of genetic medicines. At this current moment, some of them have been carried on in clinical stages, which are intriguing to keep attention on the results coming soon. In this review, we address the current trends in genetic medicines especially antisense oligonucleotides and aptamers. PMID- 9750533 TI - Multiple chemical sensitivity. PMID- 9750534 TI - The cost of treating deaf and hard-of-hearing patients. PMID- 9750535 TI - Ileoileal intussusception with a leading Meckel's diverticulum. PMID- 9750536 TI - Massive pericardial effusion in sarcoidosis. PMID- 9750537 TI - Respiratory arrest after peak expiratory flow measurement. PMID- 9750538 TI - Pediatric vision screening for the family physician. PMID- 9750539 TI - Functional endoscopic sinus surgery. AB - Functional endoscopic sinus surgery is a minimally invasive technique used to restore sinus ventilation and normal function. The most suitable candidates for this procedure have recurrent acute or chronic infective sinusitis, and an improvement in symptoms of up to 90 percent may be expected following the procedure. Fiberoptic telescopes are used for diagnosis and during the procedure, and computed tomography is used to assess the anatomy and identify diseased areas. Functional endoscopic sinus surgery should be reserved for use in patients in whom medical treatment has failed. The procedure can be performed under general or local anesthesia on an outpatient basis, and patients usually experience minimal discomfort. The complication rate for this procedure is lower than that for conventional sinus energy. PMID- 9750540 TI - Multiple chemical sensitivity syndrome. AB - Multiple chemical sensitivity (MCS) is a syndrome in which multiple symptoms reportedly occur with low-level chemical exposure. Several theories have been advanced to explain the cause of MCS, including allergy, toxic effects and neurobiologic sensitization. There is insufficient scientific evidence to confirm a relationship between any of these possible causes and symptoms. Patients with MCS have high rates of depression, anxiety and somatoform disorders, but it is unclear if a causal relationship or merely an association exists between MCS and psychiatric problems. Physicians should compassionately evaluate and care for patients who have this distressing condition, while avoiding the use of unproven, expensive or potentially harmful tests and treatments. The first goal of management is to establish an effective physician-patient relationship. The patient's efforts to return to work and to a normal social life should be encouraged and supported. PMID- 9750542 TI - Heat-related illnesses. AB - Heat-related illnesses cause 240 deaths annually. Although common in athletes, heat-related illnesses also affect the elderly, persons with predisposing medical conditions and those taking a variety of medications. Symptoms range from mild weakness, dizziness and fatigue in cases of heat edema, to syncope, exhaustion and multisystem complications, including coma and death, in cases of heat stroke. Milder heat-related symptoms can be treated with hydration, rest and removal from the hot environment. Heat stroke, a life-threatening problem, must be treated emergently. Prompt recognition is critical since rapid cooling is the cornerstone of treatment and must not be delayed. Fluid resuscitation with dextrose and normal or half-normal saline is also important. These therapies should be instituted while the patient is being stabilized. Heat illness may be prevented by recognizing which individuals are at risk, using appropriate hydration and paying attention to acclimatization and environmental conditions. Preventive care should include drinking plenty of fluids before, during and after activities, gradually increasing the time spent working in the heat and avoiding exertion during the hottest part of the day. PMID- 9750541 TI - External cephalic version. AB - External cephalic version is a procedure that externally rotates the fetus from a breech presentation to a vertex presentation. External version has made a resurgence in the past 15 years because of a strong safety record and a success rate of about 65 percent. Before the resurgence of the use of external version, the only choices for breech delivery were cesarean section or a trial of labor. It is preferable to wait until term (37 weeks of gestation) before external version is attempted because of an increased success rate and avoidance of preterm delivery if complications arise. After the fetal head is gently disengaged, the fetus is manipulated by a forward roll or back flip. If unsuccessful, the version can be reattempted at a later time. The procedure should only be performed in a facility equipped for emergency cesarean section. The use of external cephalic version can produce considerable cost savings in the management of the breech fetus at term. It is a skill easily acquired by family physicians and should be a routine part of obstetric practice. PMID- 9750543 TI - NIH panel identifies 11 principles for HIV therapy. PMID- 9750544 TI - NIH consensus panel advocates increased access to treatment for opiate addiction. PMID- 9750546 TI - An open letter to the director-general of WHO. PMID- 9750545 TI - Adduction arytenopexy: a new procedure for paralytic dysphonia with implications for implant medialization. AB - Arytenoid adduction was designed to enhance posterior glottal closure in patients with paralytic dysphonia by reproducing lateral cricoarytenoid muscle function. However this procedure can exaggerate normal medial rotation of the vocal process, because the agonist-antagonist function of the interarytenoid, lateral thyroarytenoid, and posterior cricoarytenoid muscles is not simulated. Therefore, a new adduction procedure (adduction arytenopexy) was devised to affix the arytenoid on the cricoid facet in a more optimal position for glottal sound production. The adduction arytenopexy procedure was designed on fresh cadavers. In this technique, the lateral aspect of the cricoarytenoid joint is opened widely and the body of the arytenoid is manually medialized along the cricoid facet. A specially designed single suture is then placed through the posterior cricoid and the body or the muscular process of the arytenoid to achieve 2-point fixation. This draws the arytenoid posteriorly, superiorly, and medially for precise positioning. The arytenoid is rocked internally on the cricoid facet, and suture tension is adjusted appropriately to simulate normal cricoarytenoid adduction. In the first study, the adduction arytenopexy was compared with the classic arytenoid adduction in 10 fresh cadaver larynges. The new arytenopexy procedure resulted in an average increase of 2.1 mm (p < .01) in the length of the musculomembranous vocal fold, whereas the classic arytenoid adduction did not reveal a significant change in length. Additionally, the adduction arytenopexy resulted in a consistently higher vocal fold and a more normally contoured arytenoid than the classic adduction procedure. The second study consisted of a clinical trial in which 12 patients, who presented with a widely patent posterior glottis, underwent adduction arytenopexy in conjunction with implant medialization. The procedure was successful in all patients, and there were minimal complications. In the third study, preoperative and postoperative vocal assessment measures (stroboscopic, aerodynamic, acoustic, and perceptual) were analyzed in 9 of the 12 patients. The most striking preoperative stroboscopic observation was that 8 of the 9 patients presented with an aperiodic vibrational flutter during phonation due to severe valvular incompetence. Postoperatively, all patients developed complete closure of the glottal chink and effective entrained oscillation of the vocal folds. This visual improvement in function was commensurate with comparable changes in most of the other objective and subjective measures of vocal function. The new adduction arytenopexy procedure closely simulates the biomechanics underlying normal glottal closure and cricoarytenoid adduction. In turn, complex implant design shapes are not necessary to achieve proper alignment of the arytenoid and the vocal fold. Because the arytenoid is properly positioned prior to the medialization, implants can be sized more precisely and are unencumbered by an anterior thyroid lamina suture. These procedural innovations resulted in enhanced entrained oscillation of the glottal valve and, in turn, improved laryngeal sound production. PMID- 9750547 TI - Adding to the differential. PMID- 9750548 TI - The side effects of allopurinol. PMID- 9750549 TI - Confusion about Q waves. PMID- 9750550 TI - Anterolateral Q waves in a patient with rheumatic heart disease. PMID- 9750551 TI - A man with vague rheumatic complaints. AB - A 52-year-old man presented with a four-month history of malaise, low-grade fever, decreased appetite, and a 20-pound weight loss. He complained of joint pain and swelling, proximal muscle weakness, exertional dyspnea, and a dry cough. He also noted that his fingers had turned white and then blue when chilled and red when rewarmed. He had not had pleuritic chest pain, dysphagia, dry eyes or mouth, rash, or skin photosensitivity. PMID- 9750552 TI - Case in point. Obstructed inferior vena cava. PMID- 9750553 TI - Noninvasive diagnosis of pulmonary embolism. AB - With the development of new methods for detecting pulmonary embolism, the need for angiography has been greatly reduced. Plasma D-dimer assays, lower-limb ultrasonography, and ventilation/perfusion lung scans in combination with the clinical assessment enable safe, cost-effective diagnoses. PMID- 9750554 TI - Preventing breast cancer with tamoxifen. AB - Tamoxifen offers protection against the development or recurrence of breast cancer in a range of clinical circumstances, while simultaneously helping to prevent osteoporosis and perhaps coronary heart disease. The most serious side effects of this antiestrogen are thromboembolism and accelerated development of endometrial carcinoma; they occur primarily in postmenopausal women. PMID- 9750555 TI - How HIV resists eradication. AB - Successfully treated patients with no evidence of plasma viremia may continue to harbor stable reservoirs of HIV at multiple sites, including those shielded by blood-tissue barriers. HIV may be controllable, but few believe that it can be eradicated with the limited group of agents now available. Creative new pharmacologic strategies are needed, as well as increased commitment to the development of a safe and effective vaccine. PMID- 9750556 TI - Coccidioidomycosis: an update. AB - The incidence of coccidiodomycosis in the southwestern United States has sharply increased over the last decade as a result of environmental and demographic changes. It is important to review the epidemiology, diagnostic indicators, and therapeutic options for both immunocompetent and immunocompromised patients who have a particularly high risk of infection. PMID- 9750557 TI - Direct-to-consumer advertising and the learned intermediary. PMID- 9750558 TI - Evaluation and treatment of hip pain. PMID- 9750559 TI - Hoarseness and thyroid nodules in a middle-aged woman. PMID- 9750560 TI - Confusion and knee pain after a car accident. PMID- 9750561 TI - Chronic abdominal discomfort in a 47-year-old man. PMID- 9750562 TI - Characteristics of health education among secondary schools--School Health Education Profiles, 1996. AB - PROBLEM/CONDITION: School health education (e.g., classroom training) is an essential component of school health programs; such education promotes the health of youth and improves overall public health. REPORTING PERIOD: February-May 1996. DESCRIPTION OF SYSTEM: The School Health Education Profiles monitor characteristics of health education in middle or junior high schools and senior high schools. The Profiles are school-based surveys conducted by state and local education agencies. This report summarizes results from 35 state surveys and 13 local surveys conducted among representative samples of school principals and lead health education teachers. The lead health education teacher is the person who coordinates health education policies and programs within a middle or junior high school and senior high school. RESULTS: During the study period, almost all schools in states and cities required health education in grades 6-12; of these, a median of 87.6% of states and 75.8% of cities taught a separate health education course. The median percentage of schools that tried to increase student knowledge on certain topics (i.e., prevention of tobacco use, alcohol and other drug use, pregnancy, human immunodeficiency virus [HIV] infection, other sexually transmitted diseases, violence, or suicide; dietary behaviors and nutrition; and physical activity and fitness) was > 72% for each of these topics. The median percentage of schools that tried to improve certain student skills (i.e., communication, decision making, goal setting, resisting social pressures, nonviolent conflict resolution, stress management, and analysis of media messages) was > 69% for each of these skills. The median percentage of schools that had a health education teacher coordinate health education was 33.0% across states and 26.8% across cities. Almost all schools taught HIV education as part of a required health education course (state median: 94.3%; local median: 98.1%), and more than half (state median: 69.5%; local median: 82.5%) had a written policy on HIV infection among students and school staff. A median of 41.0% of schools across states and a median of 25.8% of schools across cities had a lead health education teacher with professional preparation in health and physical education, and < 25% of schools across states or cities had a lead health education teacher with professional preparation in health education only. Across states, the median percentage of schools, whose lead health education teacher had received in-service training on certain health education topics, ranged from 15.6% for suicide prevention to 51.4% for HIV prevention; across cities, the median percentage ranged from 26.2% for suicide prevention to 76.1% for HIV prevention. A median of 19.7% of schools across states and 18.1% of schools across cities had a school health advisory council. Of the schools that received parental feedback (state median: 59.1%; local median: 54.2%), > 78% reported receiving positive feedback. INTERPRETATION: More than 75% of schools have a required course in health education to help provide students with the knowledge and skills they need to adopt healthy lifestyles. ACTIONS TAKEN: The School Health Education Profiles data are being used by state and local education officials to improve school health education and HIV education. PMID- 9750563 TI - Multistate surveillance for food-handling, preparation, and consumption behaviors associated with foodborne diseases: 1995 and 1996 BRFSS food-safety questions. AB - PROBLEM/CONDITION: In 1995, CDC, the Food and Drug Administration (FDA), and several state health departments collaboratively developed questions regarding food safety. This set of questions was used to collect data about food-handling, preparation, and consumption behaviors that have been associated with foodborne diseases in adults. These data will help characterize persons at high risk for foodborne illness and assist in developing food-safety education strategies for consumers and foodhandlers that are intended to reduce foodborne illness. REPORTING PERIOD COVERED: January 1995-December 1996. DESCRIPTION OF SYSTEM: Data were collected by using the 12 food-safety questions, which were administered with the 1995 Behavioral Risk Factor Surveillance Systems (BRFSS) in Colorado, Florida, Missouri, New York, and Tennessee, and the 1996 BRFSS in Indiana and New Jersey. In addition, data were collected in South Dakota from two of the standardized questions that deal with consumption of undercooked eggs and pink hamburgers. The BRFSS is a state-based system that surveys noninstitutionalized adults by telephone about their health behaviors and practices. RESULTS: This study included 19,356 completed questionnaires (2,461 in Colorado; 3,335 in Florida; 2,212 in Indiana; 1,572 in Missouri; 3,149 in New Jersey; 2,477 in New York; 2,110 in South Dakota; and 2,040 in Tennessee). During the previous 12 months, 50.2% of respondents reported eating undercooked eggs (95% confidence interval [CI] = 49.2-51.2); 23.8% reported eating home-canned vegetables (95% CI = 22.5-24.5); 19.7% reported eating pink hamburgers (95% CI = 18.9-20.5); 8.0% reported eating raw oysters (95% CI = 7.5-8.5); and 1.4% reported drinking raw milk (95% CI = 1.2-1.6). The prevalence of not washing hands with soap after handling raw meat or chicken and not washing a cutting board with soap or bleach after using it for cutting raw meat or chicken were 18.6% (95% CI = 17.8-19.4) and 19.5% (95% CI = 18.6-20.4), respectively. Less than half of respondents (45.4%, 95% CI = 44.2-46.6) reported seeing safe food-handling label information on raw meat products. In addition, among those persons who reported they remembered seeing the label information, 77.2% (95% CI = 76.0-78.4) remembered reading the label information, and 36.7% reported changing their meat and poultry preparation habits because of the labels (95% CI = 35.2-38.2). When population characteristics were considered in the analysis, all high-risk food-handling, preparation, and consumption behaviors were more prevalent in men than in women. Eating pink hamburgers during the previous 12 months was more commonly reported by whites (22.3%) than by blacks (6.5%). The prevalence of reported consumption of pink hamburgers during the previous. 12 months decreased with age (18-29 years: 21.8%, 30-59 years: 21.9%, and 60-99 years: 13.2%); increased with education (less than grade 12: 12.0%, high school graduate: 16.5%, and any college education: 24.0%); and increased with income (< $15,000: 11.8%, $15,000 $34,999: 17.6%, $35,000-$49,999: 22.0%, and > or = $50,000: 28.6%). INTERPRETATION: During 1995-1996, several high-risk food-handling, preparation, and consumption behaviors were common, and some were particular to specific population groups. Based on this analysis, interventions are needed to reduce the prevalence of these risky behaviors. All consumers and foodhandlers could benefit from food-safety education. ACTIONS TAKEN: Behavioral surveillance systems can provide data that identify persons or groups in which behaviors associated with foodborne diseases are more common and who are at higher risk for foodborne illness. State-specific data can assist in developing food-safety education programs and, if collected periodically, can be used to evaluate program effectiveness. PMID- 9750564 TI - Depressive symptoms of whites and African Americans aged 60 years and older. AB - Consistent with prior work, our hypothesis was that older African Americans are less likely to report dysphoria than are older Whites. Study subjects were 968 participants aged 60 years and older in Baltimore, Maryland, and 1,486 participants aged 60 years and older in the Durham-Piedmont region of North Carolina who identified themselves as African American or White and who had complete data on symptoms of depression active in the one month prior to interview, as well as several covariates thought to be related to depression. The effect of self-reported race on the endorsement of symptoms from the section on Major Depression in the Diagnostic Interview Schedule was estimated employing structural equations with a measurement model. Older African Americans were less likely to report dysphoria than older Whites, although this only achieved statistical significance by conventional standards at the Durham-Piedmont site. Older African Americans at both sites were significantly more likely to report thoughts of death. PMID- 9750565 TI - The age and job satisfaction relationship: does its shape and strength still evade us? AB - Many investigations have examined the relationship between age and job satisfaction. However, various types of relationships have been reported across studies: positive linear, negative linear, U-shaped, inverted U-shaped or inverted J-shaped, or no significant relations. Such conflicting results have left the true nature of the relationship unresolved. The present study used a large national probability sample of workers (N = 1,095) to investigate the shape and strength of the age--job satisfaction relationship. Results indicated a significant but weak positive linear age--job satisfaction relationship. That is, age failed to explain a substantial proportion of linear variance in our job satisfaction measure. This indicates that age, as a chronological variable, is not a viable predictor of job satisfaction. Future research attempting to explain age differences in job satisfaction should instead focus its attention on other more pertinent psychological variables associated with the underlying aging process. PMID- 9750566 TI - Wandering: a significant problem among community-residing individuals with Alzheimer's disease. AB - This study evaluated the frequency, predictors, and effects of wandering in a population-based sample of 193 individuals with Alzheimer's disease (AD). Although wandering occurred in subjects at all levels of cognitive impairment, analysis of variance indicated that for the group as a whole, greater frequency of wandering was associated with significantly more impairment in cognition, day to-day functioning, and behavior. Caregiver distress also increased significantly with increased frequency of wandering. Logistic regression modeling identified functional impairment and disruptive behavior problems as the strongest independent predictors of wandering occurring within the past week. Cluster analysis revealed four characteristic groups of wanderers that represented a continuum of wandering frequency, each having a unique pattern of other behavioral disturbances. Based on this analysis, we recommend further evaluation and the development of possible treatment strategies that address the individual differences found among AD patients who wander. PMID- 9750567 TI - Predictors of aggressive behaviors: a longitudinal study in senior day care centers. AB - Aggressive behaviors place elderly persons and their caregivers at risk. This study examines longitudinally the predictors of aggressive behaviors based on staff and family members' ratings of 200 community-dwelling participants from senior day care centers. The main difference between physically and verbally aggressive behaviors was found to be the role of physical and mental health. Cognitive impairment and poor quality of relationship were the main predictors of physically aggressive behaviors. Verbally aggressive behaviors were predicted mainly by depressed affect, low quality of relationships, and poor physical health. These results validate and expand prior cross-sectional research on the correlates of aggression in other populations, and guide in the development of interventions. PMID- 9750568 TI - Longitudinal and genetic effects in the relationship between pulmonary function and cognitive performance. AB - Previous studies have found cognitive deficits in patients with impaired pulmonary function, and recent data from healthy older adults suggest an association of pulmonary function with cognitive function. This 6-year longitudinal study evaluated genetic and environmental sources of covariation in the association of pulmonary function and cognitive performance. The sample included 222 Swedish twin pairs (60% women) with a mean age of 62.3 (+/- 7.7) years (age range: 40-84). Hierarchical multiple regression analyses, controlling for the effects of age, gender, and height, were employed to predict performance on cognitive tests of fluid intelligence (Digit Symbol, Block Design, Digit Span Backward) and crystallized intelligence (Information) from forced expiratory volume in one second (FEV1). Bivariate cross-twin correlations were used to evaluate the contribution of genetic and environmental factors in the association of pulmonary function and cognitive performance. Results indicated that FEV1 predicted performance on tests of fluid intelligence but not crystallized intelligence at the initial assessment and at the 6-year follow-up. Cross-twin correlational analyses indicated that genetic effects accounted for a greater share of the association of pulmonary function and cognitive performance than environmental effects, but environment also accounted for a substantial share of the covariance. PMID- 9750569 TI - Spatial cuing in a stereoscopic display: attention remains "depth-aware" with age. AB - Previous research has demonstrated that spatial attention is "depth-aware": Reaction times (RT) are greater for shifts in depth and two-dimensional (2-D) space than in 2-D space alone. This experiment examined whether the ability to focus attention at a depth location is maintained with advanced age. Twelve younger and 12 older observers viewed stereoscopic displays in which one of four spatial locations was cued. Two of the locations were at a near depth location and two were at a far depth location. When the focus of visual attention was shifted to a new location in space (because of an invalid cue), the cost in RT for switching attention (measured as the difference between RT on valid cue and invalid cue trials) was greater when observers had to switch attention between different depth locations and different locations in 2-D space than for shifts in 2-D space alone. This effect was observed for both younger and older observers, suggesting that the ability to orient attention to a depth location is maintained with age. PMID- 9750570 TI - Influences of age and perceived activity difficulty on activity recall. AB - After they performed each of a series of activities, older and younger adults were asked to rate the difficulty of the activity. Recall of the activities was later tested. Older adults tended to remember those activities they perceived to be less difficult, whereas younger adults tended to remember those activities they perceived to be more difficult. Thus, when more cognitive effort was required to perform an activity, older adults tended to have difficulty later remembering the activity. Difficult activities are hypothesized to tax limited processing resources and induce anxiety in older adults, preventing successful encoding. PMID- 9750571 TI - Age differences in attitudes toward computers. AB - It is commonly believed that older adults hold more negative attitudes toward computer technology than younger people. This study examined age differences in attitudes toward computers as a function of experience with computers and computer task characteristics. A sample of 384 community-dwelling adults ranging in age from 20 to 75 years performed one of three real-world computer tasks (data entry, database inquiry, accounts balancing) for a 3-day period. A multidimensional computer attitude scale was used to assess attitudes toward computers pretask and posttask. Although there were no age differences in overall attitudes, there were age effects for the dimensions of comfort, efficacy, dehumanization, and control. In general, older people perceived less comfort, efficacy, and control over computers than did the other participants. The results also indicated that experience with computers resulted in more positive attitudes for all participants across most attitude dimensions. These effects were moderated by task and gender. Overall, the findings indicated that computer attitudes are modifiable for people of all age groups. However, the nature of computer experience has an impact on attitude change. PMID- 9750572 TI - Barriers to voluntary organization membership: an examination of race and cohort differences. AB - OBJECTIVES: This research uses age stratification, isolation, compensatory, and ethnic community perspectives to predict differences by race in the utilization of formal organizations across cohorts. Voluntary organizations are classified into three general types: social service clubs, job-related groups, and neighborhood organizations. We hypothesize that racial differences in organizational participation will be wider for older cohorts than for younger cohorts, as a result of historical racism. Moreover, we expect the racial differences across cohorts to be greater for those organizations (i.e., social service and job-related groups) where racial barriers to membership were strongest. METHODS: We use the National Survey of Families and Households (NSFH) and logistic regression analysis to determine the predicted probabilities of membership in organizations by race, age, and type of membership. RESULTS: The results reveal higher levels of participation in organizations for young Blacks (than for young Whites). At the oldest ages, however, the race differential reverses direction for social/service and job-related organizations. For neighborhood organizations, the race differential is more stable across cohorts, consistent with expectations. DISCUSSION: We interpret these race-cohort patterns as evidence of historical discrimination that affected the oldest cohorts to a greater extent--especially for social/service and job-related organizations. PMID- 9750573 TI - Perceived self-efficacy and grandparenting. AB - OBJECTIVES: This study identifies grandparents who feel efficacious in their role and the consequences of such beliefs for actual involvement with an adolescent grandchild. METHODS: The sample of 883 grandparents comes from two related studies of rural families, the Iowa Youth and Families Project and the Iowa Single Parent Project. Our research questions are answered by testing a series of bivariate and multivariate regression models. RESULTS: Results show much variability in perceptions of being able to influence one's grandchild. Church attendance, knowledge of one's own grandparents, a farm history, a strong grandparent-parent bond, proximity, and having fewer grandchildren emerged as significant predictors of grandparents' perceptions of efficacy. Grandparents with strong self-efficacious beliefs play an active role in the lives of their grandchildren. DISCUSSION: With an increasing number of grandparents taking responsibilities for their grandchildren, a greater understanding of the experiences and resources that enhance their sense of personal efficacy in this role warrants priority in generational studies. PMID- 9750574 TI - Private health insurance coverage and disability among older Americans. AB - OBJECTIVES: This study examines the relationship between the lack of private supplemental health insurance coverage and the development of disability among adults aged 65 and older. METHODS: Data are from the baseline and six follow-up waves of the Duke Established Populations for Epidemiologic Studies of the Elderly survey (N = 4,000). Discrete-time hazard models were used to estimate the impact of insurance coverage and other risk factors on the incidence of disability among those unimpaired at baseline. RESULTS: Controlling for education, income, and other potential confounders, the odds of developing disability were 35-49% higher among those without private coverage. Insurance coverage also statistically explained part of the increased risk of disability among low-income persons. DISCUSSION: The results indicate that changes in health insurance coverage as well as in individual behaviors may be needed to reduce disability generally and disability among the socioeconomically disadvantaged, in particular. PMID- 9750576 TI - Meeting filial responsibilities in brothers-only sibling groups. AB - OBJECTIVES: This research examined how sons in brothers-only sibling groups meet the needs of their elderly parents. METHODS: 49 pairs of brothers without sisters whose parents were 74 years of age or older participated in open-ended interviews to explain how their parents' needs were met. Inductive analysis of interviews identified elements of these brothers' approaches. RESULTS: Brothers were in routine contact with and performed "masculine" services for their parents. Brothers acted independently and expected to be asked rather than volunteering help to their parents, except during times of crisis and transition. They defined their parents as self-sufficient, even when their parents' situations were precarious, and acted to reestablish their parents' independence when it was threatened. They viewed their parents' use of informal networks and formal services as appropriate. Brothers' wives provided "gender-appropriate" services. Wives' levels of involvement appeared to be a function of the quality of their relationships with their parents-in-law. Wives who helped were part of a support network. DISCUSSION: Brothers' goals of maintaining or reestablishing independence for their parents matches most parents' wishes to be independent and not to burden their children. The brothers' goal of self-sufficiency for their parents precluded their wives being the sole providers of support to their parents. PMID- 9750575 TI - Stress reduction for family caregivers: effects of adult day care use. AB - OBJECTIVES: This study reports the findings of an evaluation of the psychological benefits of use of adult day care by family caregivers assisting a relative with dementia. METHODS: The study used a quasi-experimental design in which caregivers in the treatment group used substantial amounts of services, whereas caregivers in a control group did not use day care at any point during the evaluation and only small amounts of other respite services. The evaluation was guided by the stress process model of caregiving which distinguishes between appraisals of primary stressors and well-being. RESULTS: Results after 3 months of day care use showed that the treatment group had significantly lower scores than the control group on two of the three measures of primary appraisals (overload and strain) and two of the three measures of well-being (depression and anger). Findings at one year showed that the treatment group had significantly lower scores on overload and depression than the control group. DISCUSSION: These results demonstrate that use of adult day care by caregivers of dementia patients results in lower levels of caregiving-related stress and better psychological well-being when compared to that of controls. PMID- 9750578 TI - [Apropos of intracranial pressure]. PMID- 9750577 TI - Informal exchanges with non-kin among retired sunbelt migrants: a case study of a Finnish American retirement community. AB - OBJECTIVES: This study examined informal exchanges with non-kin among retired Sunbelt migrants, with special emphasis on the impact of ethnic enclaves in generating exchanges of instrumental assistance and emotional support among elderly European Americans. METHODS: Data were collected through interviews with four samples of elders: Finnish Americans who migrated to an ethnic retirement community in Florida; European Americans who migrated to the same community but are not part of an ethnic enclave; Finnish Americans living in an age-integrated setting in Minnesota; and retired European Americans living in the same Minnesota community. RESULTS: Migrants were less likely than elders aging-in-place to report informal exchanges with non-kin. Finnish American migrants were less likely than other European American migrants to provide instrumental assistance to non-kin but were more likely to anticipate relying on informal long-term care support, regardless of proximity to kin. There were no differences in the two migrant samples in exchanges of emotional support. DISCUSSION: Evidence regarding substitution of non-kin for geographically distant kin is mixed. Results are consistent with a strategy of "banking" support, at the community level among the Finnish American migrants and at the network level among other European American migrants. PMID- 9750579 TI - [Explanatory factors for length of stay in the postoperative intensive care unit]. AB - OBJECTIVES: Because of the increasing role of post-anaesthesia care in anaesthetic activity, the explicative value of various factors on post anaesthesia length of stay (POLS) was tested. STUDY DESIGN: Retrospective study. PATIENTS: Population of 38,655 patients admitted to the post-anaesthesia care unit (PACU) between 1990 and 1994. METHOD: Multivariate analysis (semiparametric Cox model) of POLS. RESULTS: Overall 71% of the patients stayed less than four hours in PACU. Average POLS did not vary with years. Eleven variables were related to POLS, with an odds-ratio (OR) between 0.75 and 0.77 (negative link) or between 1.22 and 2.77 (positive link). A comparison of the OR for years 1990 to 1994 indicated major variations for the following three variables: neurosurgery, mechanical ventilation in PACU, low occupancy rate in the PACU at the time of discharge. CONCLUSION: POLS are linked with various factors whose changes in explicative value can be analysed chronologically. However, other methods remain necessary in order to evaluate the impact of organisational modifications, as the introduction of objective criteria for discharge. PMID- 9750581 TI - [Management of severe burns during the 1st 72 hours]. AB - Early and efficient management of severely burned patients facilitates outcome improvement. Pre-hospital care includes fluid loading with 2 mL.kg-1/% burn over the first six hours, sedation and analgesia, prevention of hypothermia and ventilatory support for either critically burned patients or facial, cervical or pulmonary burn injury. The transient stay in a general hospital before transfer to a burn centre allows extension of initial care, the critical investigation for associated injuries (intoxication, multiple trauma) and to perform initial local treatment with sterile coverage or vaseline gauze after a revised assessment of the burned skin area, and possibly escharotomies. The main aim of care in the burn centre is to control hypovolaemia and to obtain maximal tissue perfusion and oxygen delivery to burned tissues, as well as to healthy organs. To manage the burn shock (initially hypovolemic and later on hyperdynamic) catecholamines are often indicated when appropriate fluid loading remains insufficient. Mechanical ventilation is indicated in case of either a deep extensive burn over 60% of total body surface area, or facial and cervical burns or severe pulmonary burn injury from smoke inhalation, carbon monoxide intoxication, tracheobronchial thermal injury and blast injury. Because of the severity of burn-related pain, and the stimulus linked to intensive care, continuous sedation is usually required. Early surgical treatment such as escharotomies, excision and grafting, which cause significant pain as well as blood loss, and hydrotherapy, often require general anaesthesia. Burn injury can modify the volume of distribution and the pharmacokinetics of anaesthetic agents. Finally, chemical or electrical burn, radiation, associated CO intoxication or multiple trauma, as well as burn injury in infants, raise specific problems. With improvement in early intensive care, the survival rate of the most severely burned patients is obviously improving. New techniques in skin substitution will probably further improve the final outcome. PMID- 9750580 TI - [Headaches after spinal anesthesia: prospective multicenter study of a young adult population]. AB - OBJECTIVE: We assessed the occurrence of post-dural puncture headache (PDPH) in a group of young adults following spinal anaesthesia using a 24-gauge Sprotte needle. STUDY DESIGN: Prospective, multicentre, non-randomized study. PATIENTS: This 9 month-long study, included 1,122 patients less than 50 years-old, consisting of 502 women and 620 men. METHODS: Assessment of PDPH after 48 hours and 7 days. RESULTS: PDPH occurred in 0.8 percent of patients. There was no statistically significant difference in terms of age group or gender between the patients. Incidence of PDPH did not depend on type of anaesthetic solution, puncture level or ease of puncture. DISCUSSION: The use of 24-gauge Sprotte needles was associated with a low rate of puncture difficulties. Usual predisposing factors for PDPH, such as age below 50 years and female gender do no longer apply with this type of needle. The rate of puncture difficulties was low (6.7 percent), in contrast with ultra-fine 27 or 29 gauge needles, which sometimes result in puncture failure. Acceptance of the technique was excellent, as 99.38 percent of patients were satisfied. CONCLUSION: The indications of spinal anaesthesia could be extended to young patients, whatever their gender, using a non-traumatic 24-gauge Sprotte needle. PMID- 9750582 TI - [Re-expansion pulmonary edema after excision of an intrathoracic tumor]. AB - We report an acute respiratory insufficiency following the removal of a large intrathoracic fibroma (3.1 kg) in a 6 year-old child, caused by a re-expansion pulmonary oedema (unilateral oedema occurring within one hour after expansion). This oedema improved rapidly and was followed by a well-tolerated pleural effusion. This complication is due to discrepancy between a small lung and a large thoracic cavity, due to the prolonged time course of the tumor growth. These oedemas are caused by rapid lung re-expansion, the volume of the removed tumor and the depth of postoperative pleural suction. The value of positive-end expiratory pressure is discussed. PMID- 9750583 TI - [Therapeutic monitoring of teicoplanin in a severely burned patient]. AB - The authors report the study of the kinetics in serum and urine and the clinical safety of a high dose of teicoplanin administered in a 19 year-old patient with major burns (60% of body surface area, the half of which consisting of third degree burns and UBS at 150) and S aureus meticillin-resistant infection. At day 1, he was given two loading infusions of 12 mg.kg-1 teicoplanin followed by 12 mg.kg-1 per day of treatment. At all times, Cmin concentrations were below the limit value of 8 mg.mL-1. Therefore the therapeutic regimen was increased on several occasions. On days 5, 8 and 15, Cmin were measured by FPIA. Pharmacokinetic analysis was performed at day 16, (i.e., 20 mg.kg-1) and urine was also collected over at least 12 hours. At day 16, serum and urine samples were assayed by HPLC. Data were analyzed with a noncompartmental method. The duration of treatment was 20 days and no adverse events were noted. Bacteriological tests performed at the end of treatment demonstrated the elimination of the agent responsible over the infection. While pharmacokinetics were not assessed at plateau, Cmin remained very low. Vss was similar to values obtained in healthy subjects while total clearance was increased. This phenomenon was explained by the increase of total clearance and a nonrenal translesional diffusion suggested by the body surface area affected by third-degree burns. Finally, the cost of increasing doses of teicoplanin must be taken in account. PMID- 9750584 TI - [Anuric renal insufficiency caused by bilateral ureteral lithiasis]. AB - A 73-year-old man was admitted to the ICU for anuria. He reported no history of urinary disease. The abdominal roentgenography and two echographies showed an empty urinary bladder, a right ureteral calculus without dilatation of the urinary tract. Computed tomography demonstrated the presence of a left ureteral stone. Bilateral retrograde ureteroscopy and drainage allowed a rapid recovery. When the abdominal roentgenography and echography cannot explain the occurrence of anuria, the computed tomography, or better the helical CT, can demonstrate the presence of otherwise unrecognized calculi. PMID- 9750585 TI - [Coronary gas embolism after laparoscopic surgery]. AB - Gas embolism is a severe complication of laparoscopic surgery. We report two cases: one with lethal peroperative cardiac arrest from massive coronary artery gas embolism recognized during open-chest cardiac massage; the second case, also associated with coronary artery gas embolism, resulted in severe but transient abnormal left ventricular anterior wall motion, subepicardial ischaemia and injury in ECG leads V1 to V5, but unremarkable coronary arteriography and full recovery. The pathophysiology of gas embolism occurring during a laparoscopic procedure, the mechanisms of gas entry into the systemic vascular bed, the clinical, ECG, pulse oximetry, end-tidal CO2 concentration changes and alarm signs are discussed. The diagnostic value of Doppler transoesophageal echocardiography when possible is underlined, and emergency management of gas embolism is considered. PMID- 9750586 TI - [Unilateral nasal obstruction,. An unusual complication of nasotracheal intubation]. AB - Partial avulsion of the middle turbinate is an unusual complication of nasotracheal intubation, while minor nasal mucosal trauma is common. We report a case in a 25 year-old healthy-woman, diagnosed four years after nasotracheal intubation for removal of wisdom teeth under general anaesthesia, consisting in a unilateral nasal obstruction related to partial avulsion of the middle turbinate. PMID- 9750587 TI - [Biomechanics and intracranial hypertension]. AB - Biomechanics can be defined as the physical concepts exploring the effects of mechanical forces in biology. Biomechanics explores the consequences of an alteration in the steady-state normally existing between living tissues and their immediate environment. The application of this science in the field of brain physiopathology and intracranial hypertension is essential to understanding the sequence of events triggered during an alteration of the intracranial volume such as observed with intracranial pathologies. Furthermore, biomechanics allows the development of a logical and rational therapeutic approach, better adapted to the various intracerebral problems anaesthesiologists might be confronted with. PMID- 9750588 TI - [Slow pressure waves during intracranial hypertension]. AB - Intracranial pressure waves include fast waves (pulse and respiration) and slow waves. Only the latter are considered here. Since the definition of three wave types in the pioneering works of Janny (1950) and Lundberg (1960), their study of frequential characteristics shows they are included in a spectrum where three contiguous frequency bands are individualised: the B wave band (BW) between 8 x 10(-3) Hz and 50 x 10(-3) Hz; the Infra B band (IB) below 8 x 10(-3) Hz; and the Ultra B band (UB) beyond 50 x 10(-3) Hz to 200 x 10(-3) Hz. The origin of these waves is vascular and some may be physiological. They are probably generated by central neuro-pacemakers and/or cyclic phenomena of cerebral autoregulation. They are linked with slow peripheral arterial pressure waves, with biological rhythms and with biomechanics and vasomotricity in the craniospinal enclosure. They are pathological for the slowest (IB), particularly if they are plateau waves, but the physiologic-pathologic boundary is not yet established for each type of slow waves. They can cause severe consequences if they result in major cerebral perfusion pressure changes, and if they induce or worsen herniations. PMID- 9750589 TI - [Post-traumatic intracranial hypertension and biochemical disorders: cases and consequences]. AB - Increased intracranial pressure is a risk factor which may result in secondary brain damage, and affect neurological outcome in head injured patients. In case of diffuse brain lesions, elevated intracranial pressure is characterised by two important features. First, it results from vasogenic or cellular oedema, or from an increase in cerebral blood volume. Second, it is strongly associated to biochemical disorders. The latter may be considered as a direct consequence of the initial traumatic impact, mediating factors of the secondary neurological lesion and the biochemical result of cerebral ischaemia. They contribute to increased intracranial pressure and ischaemia by inducing physiological disorders and cell lesions. They also reflect the degree of cerebral ischaemia. Cerebral acidosis, free radicals and excitatory amicoacids are the main biochemical disorders implicated in this vicious circle leading to neuronal death. PMID- 9750590 TI - [Techniques for measuring intracranial hypertension]. AB - A wide variety of monitoring devices have been used for intracranial pressure measurement. The aim of this article is to present the most common devices and to assess their accuracy, stability and complications, with reference to current literature. Measurement with an intraventricular catheter remains, the reference method. However new techniques with distal measurement (fiberoptic or strain gauge) seem to be accurate, but have a higher cost. Some practical problems, such as the zero pressure reference level and the side of measurement, are also discussed. PMID- 9750591 TI - [Evaluation of ischemic repercussions of intracranial hypertension]. AB - The main risk involved in severe intracranial hypertension is, the occurrence of cerebral ischaemia, either locally during herniation or globally as a consequence of reduced cerebral perfusion pressure (CPP). Neurological features of ischaemia occur at a late stage. A continuous monitoring of brain function with EEG or evoked potential techniques, while largely used in the operating room have not been so far fully evaluated in the intensive care setting. Therefore, ischaemic criteria based on the registration of haemodynamic or metabolic data are gaining importance in management of increased intracranial pressure (ICP). Transcranial Doppler of middle cerebral arteries allows at any time the detection of a decrease in brain perfusion. An increased pulsatility index has been repeatedly demonstrated to correlate with decreased CPP. From these reports, the lower limit of autoregulation in brain injured patients appears to be much higher (70 mmHg) than previously estimated (40 mmHg). However, therapies with a cerebral vasoconstrictor impact and associated vasospasm are to be considered for a correct interpretation of Doppler data. Moreover, as a reduced cerebral blood flow is not necessarily insufficient to meet metabolic requirements, a routine insight in cerebral oxygenation and lactate production must be available. Continuous monitoring of jugular blood oxyhaemoglobin saturation (SjO2) measures the reserve of oxygen extraction and a decrease in SjO2 below 50% is considered as to indicate an impending cerebral ischaemia. Indeed, critically reduced CPP under a 70 mmHg limit is reflected by venous desaturation episodes. Increased cerebral lactate production, routinely appraisable by serial measurements of [(a v) lactate], may afford confirmation of an existing ischaemia. ICP and CPP monitoring remains the basis for intensive care surveillance during the phase of intracranial hypertension, with alarming settled at admitted critical values (ICP = 30 mmHg; CPP = 70 mmHg). As ischaemic threshold for cerebral blood flow may be different in patients and in normal experimental animals, the reliability of these critical values of ICP and CPP is uncertain. Therefore, transcranial Doppler, jugular metabolic monitoring and, as recently available, cortical tissue PO2 monitoring are mandatory for early detection and assessment of ischaemia. PMID- 9750592 TI - [Indications for monitoring intracranial pressure]. AB - A main indication for intracranial pressure monitoring is severe head trauma, where it acts as a diagnostic, prognostic and therapeutic guide. Others indications for intracranial pressure monitoring are patients with CSF circulation disturbances, whatever the cause, and various pathologies inducing intracranial hypertension, such as encephalopathies. Intracranial pressure monitoring must be associated with the measurement of mean arterial pressure, arterial and jugular venous oxygen saturation and blood flow velocity in major intracranial arteries with transcranial Doppler sonography. PMID- 9750593 TI - [Intracranial pressure monitoring in France, and French-speaking Belgium and Switzerland. Retrospective and prospective survey]. AB - OBJECTIVE: To assess the use of ICP monitoring in France and French-speaking Belgium and Switzerland. PARTICIPANTS: Neuro-intensive care unit (ICU) and neuroanaesthesia teams. METHODS: Retrospective survey (concerning the year 1995), followed by a prospective one (spread over a period of 4 months in 1996) about policy concerning: frequency, delay, duration, indications, complications and cost of ICP monitoring in their units. Other questions were: type of devices, duration of the device insertion, operators and place of operating. RESULTS: Out of the 77 units which replied, 54 regularly carried out ICP monitoring, corresponding to a total of 2,012 patients in 1995. Only 23 participated in the prospective survey (301 patients). Out of the 54 hospitals, 25 assessed the ICP in more than 50 patients per year. Head trauma was the main indication for ICP monitoring (61%), the second indication was intracranial haemorrhage (23% of patients). The most often monitored patients were young males for head trauma, whereas females were mainly monitored for intracranial haemorrhage. The delay for starting ICP monitoring was 16 hours and was maintained for 7 days in head trauma patients. Intraventricular catheter and fibreoptic devices were chosen in the same proportion, a subdural system was used in 8% of the cases. The devices were inserted by neurosurgeons in 80% of cases, anaesthetists also participated in this operation. Risk of haemorrhage was very low (0.7%), the risk of infection was 4% and always concerned intraventricular catheters. CONCLUSION: ICP monitoring is widely practised in France and French-speaking Belgium and Switzerland, especially for traumatic brain injury in young males. Anaesthetists mainly took part in the operation, especially for the fibreoptic devices. PMID- 9750594 TI - [Cerebral hemodynamics and intracranial hypertension]. AB - Various cerebral aggressions, either primary or secondary, can lead to the development of raised intracranial pressure. The presence of an elevated intracranial pressure often results in cerebral ischaemia/hypoxia and, eventually, neuronal death. In face of this cascade of events, several therapeutic approaches have been suggested. Two management concepts for patients with raised intracranial pressure have retained the most attention in recent years: the first suggests a therapeutic increase in cerebral perfusion pressure with the objectives to improve perilesional collateral perfusion and decreased cerebral blood volume, and consequently intracranial pressure in areas where autoregulation is preserved. The second concept supports the diminution in perilesional capillary pressure with the aim of decreasing vasogenic oedema. Although these two concepts are antagonistic and cannot be used simultaneously, they are probably complementary in the sequence of therapeutic events of patients experiencing severe head injury. This article reviews these therapeutic concepts and their clinical applications. PMID- 9750595 TI - [The internal environment and intracranial hypertension]. AB - Intracranial pressure depends on cerebral tissue volume, cerebrospinal fluid volume (CSFV) and cerebral blood volume (CBV). Physiologically, their sum is constant (Monro-Kelly equation) and ICP remains stable. When the blood brain barrier (BBB) is intact, the volume of cerebral tissue depends on the osmotic pressure gradient. When it is injured, water movements across the BBB depend on the hydrostatic pressure gradient. CBV depends essentially on cerebral blood flow (CBF), which is strongly regulated by cerebral vascular resistances. In experimental studies, a decrease in oncotic pressure does not increase cerebral oedema and intracranial hypertension (ICHT). On the other hand, plasma hypoosmolarity increases cerebral water content and therefore ICP, if the BBB is intact. If it is injured, neither hypoosmolarity nor hypooncotic pressure modify cerebral oedema. Therefore, all hypotonic solutes may aggravate cerebral oedema and are contra-indicated in case of ICHT. On the other hand, hypooncotic solutes do not modify ICP. The osmotic therapy is one of the most important therapeutic tools for acute ICHT. Mannitol remains the treatment of choice. It acts very quickly. An i.v. perfusion of 0.25 g.kg-1 is administered over 20 minutes when ICP increases. Hypertonic saline solutes act in the same way, however they are not more efficient than mannitol. CO2 is the strongest modulating factor of CBF. Hypocapnia, by inducing cerebral vasoconstriction, decreases CBF and CBV. Hyperventilation is an efficient and rapid means for decreasing ICP. However, it cannot be used systematically without an adapted monitoring, as hypocapnia may aggravate cerebral ischaemia. Hyperthermia is an aggravating factor for ICHT, whereas moderate hypothermia seems to be beneficial both for ICP and cerebral metabolism. Hyperglycaemia has no direct effect on cerebral volume, but it may aggravate ICHT by inducing cerebral lactic acidosis and cytotoxic oedemia. Therefore, infusion of glucose solutes is contra-indicated in the first 24 hours following head trauma and blood glucose concentration must be closely monitored and controlled during ICHT episodes. PMID- 9750596 TI - [Effects of anesthetic agents on intracranial pressure]. AB - Barbiturates, etomidate and propofol decrease cerebral blood flow (CBF), mediated by a decrease in cerebral metabolism, thus decreasing intracranial pressure (ICP). As the reduction in CBF is secondary to a decrease in cerebral metabolism, these agents will have little effect on CBF or ICP in patients without active cerebral metabolic activity. Ketamine is usually not administered for the anaesthetic management of patients at risk of intracranial hypertension because of the reported increases in cerebral metabolism, CBF and ICP. The increase in CBF, however, may be partly mediated by a sympathetically induced increase in blood pressure and partly by a simultaneous increase in PaCO2 in spontaneously breathing patients. More recent studies report no increase in ICP or flow when ventilation is controlled or when other agents are associated. There is renewed interest in ketamine because it blocks excitatory amino acid receptors in the brain. Synthetic opioids including fentanyl, sufentanil, and alfentanil have been reported to cause an increase in ICP in patients with various intracranial lesions. When blood pressure was supported, no clinically relevant increase in ICP or flow velocity with alfentanil or sufentanil was observed. Thus, the increase in ICP reported with these agents may be related to the compensatory autoregulation-mediated vasodilation, underscoring the importance of administering these agents carefully to avoid systemic hypotension. Halothane consistently increases CBF and should not be used in patients with increased ICP. In contrast, isoflurane does not cause increase in CBF at concentrations below 1 to 1.5 MAC, although the effects on cerebral blood volume are less clear. Desflurane and sevoflurane have similar effects. CO2 reactivity is preserved with all inhaled agents. In patients with increased ICP however, it would be preferable to avoid these agents or to administer very low doses. PMID- 9750598 TI - [Importance of esophageal Doppler in anesthesia]. PMID- 9750597 TI - [Treatment of intracranial hypertension in the case of severe craniocerebral injuries]. AB - More than 50% of severely head-injured patients develop increased intracranial pressure, risking exacerbating ischaemic insults to the already injured brain. In approximately 10% of these cases, intracranial pressure may become unresponsive to medical or surgical treatment, with a resulting mortality of over 90%. The main emphasis should be on full intensive care, based on the prophylaxis of the devastating effects of secondary insults to the injured brain. Specific treatment should be directed towards controlling intracranial pressure and maintaining a cerebral perfusion pressure over 70 mmHg, while avoiding, where feasible, treatment modalities at risk of exacerbating cerebral ischaemia. Recently, an algorithm for treating intracranial hypertension under three different therapeutic situations has been suggested, based on the successive application of effective agents with increasing associated risks. Therapeutic modalities of this protocol are discussed. PMID- 9750599 TI - [Presentation of a method for tracheobronchial aspiration taking into account the rules for asepsis]. PMID- 9750600 TI - [Target-controlled inhalation anesthesia or computer-controlled quantitative inhalation anesthesia]. PMID- 9750601 TI - [Effects of CO2 and adrenaline on 1% lidocaine in axillary block]. AB - OBJECTIVES: To compare lidocaine hydrocarbonate and lidocaine hydrochloride, with and without adrenaline, in the axillary block obtained with a neurostimulator. STUDY DESIGN: Prospective, randomized, double blind study. PATIENTS: Sixty-six patients undergoing surgery of the upper limb under axillary block, allocated into four groups. METHODS: The criteria for evaluation were: onset time, duration and quality of sensory and motor blockades, and blood concentrations of lidocaine in 39 patients. In all patients musculocutaneous, radial, median and ulnar nerves were stimulated and the volume of local anaesthetic administered was 25 mL per square meter of body surface. Group 1 received lidocaine hydrocarbonate 1% (n = 17), group 2, lidocaine hydrocarbonate 1% with adrenaline 1/200,000 (n = 17), group 3, lidocaine hydrochloride 1% (n = 16) and group 4, lidocaine hydrochloride 1% with adrenaline 1/200,000 (n = 16). RESULTS: No significant inter-group differences were found concerning sensory and motor blockades and onset time. The duration of analgesia was longer in groups CO2 + A and HCL + A. The lidocaine blood concentrations were globally lower in group HCL + A. CONCLUSIONS: Considering the cost/benefit ratio and the absence of clinical benefits of lidocaine hydrocarbonate, lidocaine hydrochloride should be preferred. PMID- 9750602 TI - [Prevention of arterial hypotension during spinal anesthesia using intramuscular ephedrine in older people]. AB - OBJECTIVE: To assess the efficacy of intramuscular ephedrine for prevention of hypotension following subarachnoid block (SB) in the elderly. STUDY DESIGN: Prospective, randomized double blind study vs placebo. PATIENTS: Twenty patients, aged 60 years or more, of physical class ASA 2 or 3, scheduled for surgical fixation of fractured neck of femur under SB, allocated into two groups of ten each. METHODS: After oral premedication with hydroxyzine 50 mg, 90 min before surgery, and preloading with cristalloid solution 10 mL.kg-1, the subarachnoid space was punctured with the patient in lateral position using a 22 Gauge spinal needle at the L3-L4 or L4-L5 interspace. Patients were given 0.5% hyperbaric bupivacaine intrathecally, according to body weight. Patients in ephedrine group received intramuscular ephedrine 30 mg immediately after SB. Patients in placebo group received 1 mL of intramuscular saline immediately after SB. When blood pressure decreased below 100 mmHg repeated bolus of ephedrine 6 mg were given intravenously. RESULTS: Patients in both groups experienced a significant decrease in systolic pressure after SB, the decrease being significantly greater in the placebo group. CONCLUSION: Prophylactic intramuscular ephedrine is effective to prevent hypotension associated with SB in the elderly. PMID- 9750603 TI - [Combination of Emla cream and nitrous oxide for venous cannulation in children]. AB - OBJECTIVE: To assess the efficacy of an combination of Emla cream and N2O for venous cannulation in children. STUDY DESIGN: Prospective, randomized, double blind trial. PATIENTS: The study included 75 unpremedicated children, aged 3 months to 5 years, ASA physical class I and II, undergoing an elective surgical procedure, randomized into three groups. METHODS: In group I and III, children received Emla cream one hour before entering the theatre. In group II, children received a placebo. Children of group I and III also inhaled 50 vol% nitrous oxide in oxygen and those of groupe II 100 vol% oxygen, 3 min prior and during venous cannulation. A blinded observer recorded the following items: pain assessment with CHEOPS scoring, conditions of venous puncture and behaviour of children. Heart rate, blood pressure and oxygen saturation were assessed at three timepoints: before, 3 min after facial mask application and following venous cannulation. RESULTS: There were non significant differences between the three groups for the conditions of venous cannulation. The CHEOPS score was better in group I (7[4-11]), compared to group II (10[6-13]; P < 0.01) and to group III (9[6-12]; P < 0.01). CONCLUSION: Emla cream combined with nitrous oxide is effective for venous cannulation in providing satisfactory analgesia and in controlling anxiety elicited by the vision of needle. PMID- 9750604 TI - [Interviews with families of organ donors: analysis of motivation for acceptance or refusal of donation]. AB - OBJECTIVE: The reasons for organ donation acceptance or refusal are still unclear. This study analysed the influence of the circumstances of the conversations with the relatives of brain dead patients on their consent for organ donation. STUDY DESIGN: Prospective study. MATERIAL: The analysis included 41 questionnaires collected over nine months in one organ harvesting centre and focusing on the circumstances of death, the conditions of the conversations and the reasons for acceptance or refusal. METHODS: Questionnaire filled in by the physicians after the interviews of the relatives of brain dead patients. RESULTS: The refusal rate was higher (54 vs 21%) when only one physician participated in the conversation, when more than two relatives had to decide (42 vs 24%), when conversations took place during night or when the request for organ donation followed immediately the announcement of death (43 vs 20%). Most often the relatives gave their decision within minutes following the request. CONCLUSION: The circumstances of conversation with families play an essential role in their decision-making. A written guideline implementation for these conversations would probably be beneficial for the decisions of families in favour of organ donation. PMID- 9750606 TI - [Benzodiazepine withdrawal presenting as pseudo-surgical abdominal pain]. AB - A 39-year-old patient was admitted to the emergency department for acute abdominal pain. Physical examination showed a peritoneal syndrome. However, CT scan, Doppler and blood analysis were unremarkable. As the patient had a history of auto-medication with benzodiazepines at high doses, a withdrawal syndrome was considered. An intravenous administration of 3 mg of midazolam determined the relief of all symptoms in a few minutes. PMID- 9750607 TI - [Ehlers-Danlos syndrome type III and pregnancy: labor analgesia]. AB - We report the use of obstetrical extradural analgesia in a primigravida with Ehlers-Danlos syndrome type III. Management focused on the possible vascular tissue involvement and the risk of bleeding during the access into the extradural space. The procedure was performed after ruling out the anomalies in the dermis and its vasculature. PMID- 9750605 TI - [Role of non-steroidal anti-inflammatory agents in the perioperative period. Usefulness and limitations]. AB - Nonsteroidal antiinflammatory drugs (NSAIDs), including various chemical families of drugs, inhibit prostaglandin synthesis and act on the central nervous system. Prostaglandins are involved in regulation of regional circulations, cell turn over in the gastrointestinal tract, and in primary haemostasis. The patterns of action of NSAIDs result in analgesic properties, but also in adverse effects. NSAIDs are increasingly used perioperatively, alone or associated with opioids or local anaesthetics, because of their analgesic and opioid sparing properties. Some of their adverse effects, especially ischaemic acute renal failure and gastrointestinal complications, can be life-threatening, and increased haemorrhagic risk is an issue for spinal or epidural anaesthesia in patients taking aspirin. Safe use of NSAIDs is possible in consideration of contraindications (elderly patient, hypovolaemia, cirrhosis, congestive heart failure, renal failure, active gastrointestinal ulcer, bleeding diathesis, pregnancy), and requires close monitoring of renal function if they must be used in patients at risk for renal failure. NSAIDs are not ulcerogenic in the short term in healthy subjects. They must be used with caution in patients with a preexisting haemostatic defect or undergoing haemorrhagic surgical procedures. PMID- 9750608 TI - [Chylothorax complicating a subclavian puncture controlled by positive-pressure expiratory ventilation]. AB - We report a case of traumatic chylothorax which occurred after a right subclavian vein catheterisation. Chyle output exceeded 4 L.day-1 despite a continuous drainage of the pleural space, cessation of oral intake and mechanical ventilation. It was cured by addition of PEP to ventilation. The various causes and therapeutic approaches are reviewed. PMID- 9750609 TI - [Fatal hepatitis in a young child: probable role of halothane]. AB - A 3-year-old boy, who underwent multiple anaesthetics including halothane in a short period of time, developed 3 days after the last operation abdominal pain, jaundice and fever. Laboratory tests showed hepatic failure, with cytolysis, cholestasis and eosinophilia. Tests for hepatitis A, B, C, CMV and EBV were negative. No other causes of postoperative jaundice were identified. Despite symptomatic treatment, the child died 5 days after the last anaesthetic. Post mortem liver biopsy showed massive hepatic necrosis. The authors discuss factors increasing the risk for halothane-hepatitis, especially multiple exposures. PMID- 9750610 TI - [PhysioFlex: a target-controlled self-regulating closed-circuit inhalation anesthesia regulator]. AB - Physi Flex is the first commercially available apparatus capable for quantitative, or self-regulating target controlled inhalational anaesthesia, with a totally closed circuit, in adults and children. The fresh gas supply to the circuit is intermittent, automatically regulated by continuous monitoring of the volume and composition of the gas mixture in the breathing circuit. The circle system includes, instead of the two conventional one way valves, a blower creating a continuous unidirectional flow at 70 L.min-1. In addition to the CO2 absorber it contains an absorber with carbon, absorbing the anaesthetic vapour when switched into the circuit. The ventilator consists of four ventilating chambers, each one with a membrane separating the patient and the motor compartments. The displacement of the membranes generates and measures the tidal volume. Automatic ventilation is achieved by electric valves and motor gas, and manual ventilation using a bag. Spontaneous ventilation is also possible. The machine is operated via a computer with selects the number of ventilating chambers (one, two or four), and the tidal volume between 50 and 2,000 mL, depending on age, gender and weight of the patient. The computer maintains the gas volume and the gas and vapour concentrations at their preset values. The O2 flow and consumption, the N2O flow and uptake, FICO2 and FETCO2, FI and FET of the volatile anaesthetic, all other important data are displayed in a numerical and graphical form on a color screen and registered for a delayed analysis. The end tidal concentration of the volatile anaesthetic drives a stepmotor with a syringe containing the selected volatile anaesthetic agent with is directly injected into the breathing circuit where it is vaporized. Therefore the concentration of the anaesthetic vapour can be instantaneously increased with this injector at induction and lowered at end of anaesthesia with the carbon absorber, and the fresh gas consumption is significantly decreased. PMID- 9750612 TI - [Scheduled autologous transfusion and surgery of an hepatic hydatid cyst]. PMID- 9750611 TI - [Pre-hospitalization management of evolving myocardial infarction. 2d conference of south-eastern region emergency medicine experts]. PMID- 9750613 TI - [Anaphylactoid reactions during anesthesia in a satellite hospital]. PMID- 9750614 TI - [Intracameral anesthesia for cataract surgery]. PMID- 9750615 TI - [Salvage transtracheal ventilation]. PMID- 9750616 TI - [Ambulatory anesthesia practice in a University Hospital Center: what is done; what could be done. Ambulatory Anesthesia Study Group]. AB - OBJECTIVES: To estimate the number and type of patients, who could be managed on a day-care basis in a University Hospital. Cases of ambulatory anaesthesia (AA) which could be managed in optimal conditions and current AA practice. To assess patients' opinion on inpatient or outpatient practices. STUDY DESIGN: Prospective survey over 8 weeks in 21 medical units potentially concerned by ambulatory practice. PATIENTS: Series of 1,396 patients undergoing an operation included in a previously established list of outpatient procedures. METHOD: A questionnaire was completed for each patient, from preoperative anaesthesia consultation until discharge. Thereafter patients were contacted after home discharge by telephone or mail. RESULTS: In the group of the 1,396 selected patients, 301 (22%) were contra-indicated for AA. In optimal conditions AA was indicated in 20% of all patients treated in these units. At the time of the survey there were 285 outpatients (26% of potential AA, 5% of all patients). Complications were uncommon. Ninety-eight percent of them would choose day surgery again. Among the 810 inpatients, 279 responded: 53% would prefer to be an ambulatory patient. CONCLUSION: At the time of this survey only one fourth of the possible oupatients for AA had in fact been treated on a day-case basis. Its development requires an improvement of the structures, team organisation and patients' information. PMID- 9750617 TI - [Economic impact of a prescription protocol in an intensive care unit]. AB - OBJECTIVE: To assess the economic impact of a prescribing protocol for i.v. fluid therapy and artificial nutrition. STUDY DESIGN: Comparative study, before and during use of the protocol. PATIENTS: The study included 555 ICU patients allocated into two groups, before and after starting with the protocol. The groups were comparable for number, pathologies, age, severity score, duration of ICU stay, incidence of nosocomial infections, mortality rate. METHOD: In February 1995, a written literature-based prescribing protocol for fluid therapy (hydroxyethylstarch and albumin), and artificial nutrition (enteral nutrition as first-line therapy) was devised. A cost analysis was made for two 6-month periods: before (August 1994 to January 1995) and after start of protocol (February to July 1995). RESULTS: The prescription of albumin and hydroxyethylstarch decreased (by 33 and 58% respectively), whereas administration of Ringer lactate and gelatine solutes increased simultaneously. This induced a cost saving of 15,000 FF (a 20% decrease in cost). The reduction of parenteral nutrition in favour of early enteral nutrition induced a cost saving of 56,000 FF (31% decrease in cost). CONCLUSION: Our prescribing protocol generated a cost saving of 9% of the pharmaceutical budget and decreased the cost-benefit ratio of our ICU. PMID- 9750618 TI - [Tracheal intubation in prehospital resuscitation: importance of rapid-sequence induction anesthesia]. AB - OBJECTIVE: To investigate complications of emergency endotracheal intubation (EEI), possibly facilitated by rapid-sequence induction, in the prehospital critical care setting: 1) the difficulty of intubation; 2) the cardiorespiratory consequences of intubation; 3) the relationship between the occurrence of complications and prognosis. STUDY DESIGN: Prospective non randomized, open study. PATIENTS: All patients treated over a 5-month period by a physician-manned ambulance service and requiring EEI. METHODS: Patients were allocated either in with cardiac arrest (CA) group or a group with maintained spontaneous circulation (SC). Difficulty of intubation was assessed by the number of attempts. RESULTS: Two hundred and twenty-four consecutive EEI were carried out by physicians (46%) and residents (38%) not trained in anaesthesia, anaesthetists (8%), or nurse anaesthetists (7%). Trachea was intubated after a maximum of three attempts in all patients. Success rate at the first attempt was 91%. It was 92% in CA patients (n = 76) and 90% in SC patients (P = 0.59). Anaesthetic induction, with (n = 112) or without (n = 12) succinylcholine, was used to facilitate 84% of intubations in SC patients. Complications occurred in 30 patients (20%). There was no relationship between the latter and hospital mortality, duration of ventilatory support, duration of stay in the intensive care unit. CONCLUSION: In this study, EEI in SC patients was frequently facilitated by rapid sequence induction and was associated with a high success rate at the first attempt, as in CA patients. Morbidity was low. All physicians involved in emergency airway management should be skilled in this technique. PMID- 9750619 TI - [Hypothermia in traumatology]. AB - Basing on the experience of the Chamonix hospital team which managed in six years 89 cases of hypothermia in trauma patients, this article reviewed the literature concerning the association hypothermia-trauma. Shock is a major triggering factor. The deleterious effects of hypothermia on the outcome is due to inadequate cardiorespiratory adaptation to shock and to increased bleeding. Although a few articles reported a beneficial effect of hypothermia in head trauma, further studies are required to assess the value of deliberate hypothermia in such patients. Restoration of a satisfactory haemodynamic activity is a priority and most often requires surgery. The rewarming manoeuvres should be initiated early and always be preventive. They are active, internal and rapid in case of haemodynamic instability and when the central temperature is below 32 degrees C. It can be more progressive and less invasive in other cases. During recovery from anaesthesia the patient must be closely monitored. In spite of a possible protecting effect, hypothermia remains an aggravating factor in traumatology and must therefore be either prevented or amended. PMID- 9750620 TI - [Anesthesia ventilators]. AB - OBJECTIVE: To review anaesthesia ventilators in current use in France by categories of ventilators. DATA SOURCES: References were obtained from computerized bibliographic search. (Medline), recent review articles, the library of the service and personal files. DATA SYNTHESIS: Anaesthesia ventilators can be allocated into three groups, depending on whether they readminister expired gases or not or allow both modalities. Contemporary ventilators provide either constant volume ventilation, or constant pressure ventilation, with or without a pressure plateau. Ventilators readministering expired gases after CO2 absorption, or closed circuit ventilators, are either of a double- or a single-circuit design. Double-circuit ventilators, or pneumatical bag or bellows squeezers, or bag-in bottle or bellows-in-bottle (or box) ventilators, consist of a primary, or driving circuit (bottle or box) and a secondary or patient circuit (including a bag or a bellows or membrane chambers). Bellows-in-bottle ventilators have either standing bellows ascending at expiration, or hanging bellows, descending at expiration. Ascending bellows require a positive pressure of about 2 cmH2O throughout exhalation to allow the bellows to refill. The expired gas volume is a valuable indicator for leak and disconnection. Descending bellows generate a slight negative pressure during exhalation. In case of leak or disconnection they aspirate ambient air and cannot act therefore as an indicator for integrity of the circuit and the patient connection. Closed circuit ventilators with a single circuit (patient circuit) include a insufflating device consisting either in a bellows or a cylinder with a piston, operated by a electric or pneumatic motor. As the hanging bellows of the double circuit ventilators, they generate a slight negative pressure during exhalation and aspirate ambient air in case of leak or disconnection. Ventilators not designed for the readministration of expired gases, or open circuit ventilators, are generally stand-alone mechanical ventilators modified to allow the administration of inhalational anaesthetic agents. PMID- 9750621 TI - [Obstructive renal insufficiency caused by amoxicillin crystalluria]. AB - A 76-year-old woman was admitted to the ICU for a meningitis with rhombencephalitis due to Listeria monocytogenes. The treatment included amoxicillin (250 mg.kg-1.day-1) and gentamicin (3 mg.kg-1.day-1 over 6 days). Neurological outcome was favourable. However at the 14th day, an acute renal failure occurred, following macroscopic haematuria and milkiness urine. CT scan and sonography confirmed the diagnosis of obstructive renal failure with bilateral ureteral obstruction. Crystalluria caused by amoxicillin was suspected. Endoscopic ureteral insertion of double-J catheters permitted the recovery of a normal renal function. PMID- 9750622 TI - [Selenium deficiency favors the appearance of heart failure after multiple injury]. AB - A 47-year-old multiple trauma patient, experiencing a C. Difficile colitis with diarrhoea, developed diffuse oedema with peritoneal and pleural effusion due to global heart failure. Selenium deficiency, reported in trauma patients, may explain the occurrence of cardiomyopathy. The role of selenium in cardiac dysfunction and the various situations inducing a selenium deficiency are discussed. PMID- 9750623 TI - [Perioperative antibiotic prophylaxis practice of French anesthesiologists and resuscitators: results of a national survey]. AB - Antibiotics are the most prescribed drugs in French hospitals in one third of cases they are used for antiobiotic prophylaxis in surgery. In spite of the guidelines for antibioprophylaxis produced in the last years, their prescription patterns remain still often inappropriate. This survey aimed to assess whether the prescription of antibiotics for prophylaxis by French anaesthetits complied with the French recommendations for antibioprophylaxis in surgery. It focused on the recommended agents, the time of the first injection, the duration of treatment. A sample of 1,473 anaesthetists participated in the survey. In 93% of cases, the first injection of the antibiotic took place at anaesthesia induction, as specified by the recommendations. Cephalosporins of the first and second generation were often administered, as well as the association amoxicillin clavulanic acid. In contradiction with the recommendations, the cephalosporins of the third generation were widely prescribed in digestive and urological surgery, and the quinolones in urology and ophtalmology. The duration of treatment was restricted to 48 hours by 94% of anaesthetists. However there was a strong tendency to prolonge it in immunodepressed patients and in case of major surgery. This survey showed disparities between the French recommendations for antibioprophylaxis in surgery and the prescription patterns of anaesthetists. The lack of compliance occurred mainly for recent cephalosporins and treatment duration of over 48 hours. It is concluded that a stronger adherence to the principles of antibioprophylaxis is required in surgical patients. PMID- 9750624 TI - [Dental accidents in relation to general anesthesia. Experience of mutual medical insurance group]. AB - This article reviewed retrospectively 511 cases of dental damage reported to the French group for medical insurances from 1990 to 1995, and representing 40% of all accidents related to anaesthesia. The mean incidence rate was 9.5 accidents/100 anaesthetists/year and tended to decrease over time. The part of cases qualified as being linked to a fault was high and mainly due to the lack of dental examination and of informed consent during the preanaesthetic assessment. Although the mean cost of a dental accident is low compared with other anaesthetic accidents, the global cost is substantial considering their high incidence. PMID- 9750625 TI - [Neurostimulation under general anesthesia and peripheral nerve injuries]. PMID- 9750626 TI - [Blindness and eclampsia]. PMID- 9750627 TI - [A dramatic selective intubation]. PMID- 9750628 TI - [Percutaneous tracheotomy by dilatation: the Griggs or the Ciaglia technique?]. PMID- 9750629 TI - [Atrial fibrillation and acute alcohol intoxication]. PMID- 9750630 TI - [Perianesthesia anaphylactic shock: management]. PMID- 9750631 TI - [What is the role of droperidol in anesthesia-recovery in 1997?]. PMID- 9750632 TI - [Blood patch]. PMID- 9750633 TI - [A challenge in neuroresuscitation: secondary cerebral lesions of systemic origin]. PMID- 9750634 TI - [Remifentanyl]. PMID- 9750635 TI - [Centralized information system of data on anesthesia]. PMID- 9750636 TI - [Latex allergy]. PMID- 9750638 TI - [Anesthesia consultation and the pre-anesthesia visit]. PMID- 9750639 TI - [Locoregional anesthesia in children]. PMID- 9750640 TI - [Apropos of medical gas distribution circuits]. PMID- 9750641 TI - [Pressure control in medical gas distribution systems]. AB - OBJECTIVE: To assess whether the pressure gauges at the downstream part of pressure regulators are accurate enough to ensure that pressure in O2 pipeline is always higher than in Air pipeline and that pressure in the latter is higher than pressure in N2O pipeline. A pressure difference of at least 0.4 bar between two medical gas supply systems is recommended to avoid the reflow of either N2O or Air into the O2 pipeline, through a faulty mixer or proportioning device. STUDY DESIGN: Prospective technical comparative study. MATERIAL AND METHODS: Readings of 32 Bourdon gauges were compared with data obtained with a calibrated reference transducer. Two sets of measurements were performed at a one month interval. RESULTS: Pressure differences between Bourdon gauges and reference transducer were 8% (0.28 bar) in average for a theoretical maximal error less than 2.5%. During the first set of measurements, Air pressure was higher than O2 pressure in one place and N2O pressure higher than Air pressure in another. After an increase in the O2 pipeline pressure and careful setting of pressure regulators, this problem was not observed at the second set of measurements. DISCUSSION: Actual accuracy of Bourdon gauges was not convenient enough to ensure that O2 pressure was always above Air pressure. Regular controls of these pressure gauges are therefore essential. Replacement of the faulty Bourdon gauges by more accurate transducers should be considered. As an alternative, the increase in pressure difference between O2 and Air pipelines to at least 0.6 bar is recommended. PMID- 9750642 TI - [Evaluation of acute pain in prehospital medicine]. AB - OBJECTIVE: To evaluate acute pain in prehospital setting. STUDY DESIGN: Prospective survey. PATIENTS: All eligible patients during a 3-month-period, excepted children less than 10-year-old. METHOD: Pain intensity was evaluated by verbal rating scale with 5 points (VRS), visual analog scale (VAS), demand for antalgics by the patient and the relief obtained. These data were collected at the beginning (T0) and the end (Tend) of medical management. Analgesic treatments were let at the physician's choice. RESULTS: A series of 255 patients were included (mean age 58 +/- 1.5 SEM, sex-ratio 57M/43F). Among them, 42% experienced pain at VRS. VAS could be used in 60% of patients. VRS evaluated by the patient was correlated to the VAS (P < 0.001). Among those with significant pain (defined by a VAS > or = 30 mm), only 31% asked for analgesia and 64% received analgesics. Pain scales (VRS and VAS) were significantly improved (P < 0.001) at the end of the medical management, except for patients who did not receive any treatment. However, mean VAS was still above 30 mm, even in patients receiving analgesics. Only 49% of patients expressed a good relief at the end of the medical management. CONCLUSION: Acute pain is frequently observed in prehospital emergency medicine. Pain scales such as VRS and VAS are used easily and convenient for the assessment of pain intensity in this context. However, even if pain is correctly evaluated, it is still inadequately treated. The reasons of these inadequacies must be assessed and corrected with pain treatment protocols including opioids. PMID- 9750643 TI - [Incidence of myocardial lesions after vascular surgery: diagnosis by troponin Ic]. AB - OBJECTIVE: To compare the incidence of myocardial damages diagnosed following vascular surgery using the cardiac troponin I measurement technique and conventional methods. STUDY DESIGN: Prospective epidemiological study. PATIENTS: Fifty-four patients who underwent surgery for either aneurysmal disease in 28 cases or occlusive aortic disease in 26 cases. METHODS: Plasma concentration of cardiac troponin I (significant at a concentration > 1.5 ng.mL-1) was measured by immunoenzymofluorimetry on the second and fifth postoperative days. Conventional monitoring methods included daily electrocardiogram (ECG), enzymatic assay of total-PCK, and measurement of plasma levels of the MB isoenzyme of phosphokinase creatine (MB-PCK) (significant at > 1 ng.mL-1 and RI > 1.5). RESULTS: The cardiac troponin I measurement technique allowed the diagnosis of minor myocardial damages during the postoperative period in five patients, whereas with the conventional methods (clinical signs. ECG, and MB-PCK) only three myocardial lesions were diagnosed. CONCLUSION: The cardiac troponin I measurement technique allows diagnosis of minor myocardial damages following vascular surgery. Conventional methods underestimate the incidence of these damages. PMID- 9750644 TI - [Anesthesia and non-conventional transmissible agents (or prion diseases)]. AB - The transmissible spongiform encephalopathies (TSE) represent a group of neurodegenerative diseases with lethal outcome. They include Creutzfeldt-Jakob disease (CJD) and kuru, among others in humans, scrapie in sheep and spongiform encephalopathy in cattle (bovine spongiform encephalopathy: BSE). Some are autosomal dominant disorders like CJD, Gerstmann-Straussler-Scheinker disease (GSS), with point mutation of the prion protein gene. Most of these diseases are idiopathic rather than sporadic, latrogenic CJD could be obtained by central inoculation (neurosurgical instruments or dura mater grafts) or by peripheral inoculation (pituitary hormone therapy). A new variant clinicopathological type of CJD (nvCJD) has been reported. The nvCJD has strain characteristics distinct from other types of CJD, close to those of BSE transmitted (studies with intracerebral inoculation), consistent with BSE being the source of this new disease. All of these spongiform encephalopathies (SE) are characterized by spongiform degeneration of the brain, reactive gliosis in the cortical and subcortical gray matter, neuronal loss and presence of the abnormal isoform of the cellular prion protein (PrPc). In prion disease, PrPc undergoes conformational changes involving a shift from alpha-helix to beta-sheet structure. These neurologic lesions are characterized by major variations from case to case. Neuropathological studies in sporadic CDJ have emphasized phenotypic variations. Clinical presentation with a wide spectrum of manifestations is a rapidly progressive dementia, associated with myoclonus or akinetic mutism and cortical blindness. The clinical course is atypical and when the characteristic triphasic abnormal EEG of CJD is absent, there is an urgent need for a premortem diagnostic test. Histopathological examination of a brain biopsy carries a risk of major morbidity and may miss the site of disease. The 14 3-3 immunoassay of cerebrospinal fluid strongly supports a diagnosis of CJD. Western blot analysis of human tonsil biopsy may allow an early or preclinical diagnosis. It has been suggested that CJD might be transmitted by blood products derived from patients with CJD during the prodromal stage, although CJD linked aetiologically to blood transfusion has not been demonstrated. In animal studies, intracerebral inoculation of infected cells has been associated with development of disease, but never after peripheral inoculation into the blood stream. For the most part of conformational changes of PrPc, the remarkable resistance of the infectious agent (PrP alone or combined) to ordinary sterilising procedures is a major problem. Because of this resistance, current recommendations are to recognize patients at risks and to use disposable medical devices. This is particularly true in anaesthesia during endotracheal intubation, spinal anaesthesia, and to a lesser extent with peripheral nerve blocks. All instruments used for patients with CJD must be destroyed. The economic consequences of these measures have highlighted the essential importance of an early diagnosis. PMID- 9750645 TI - [Peridural abscess complicating spinal anesthesia in a diabetic patient]. AB - Infectious complications of spinal or epidural anaesthesia are rare, particularly after spinal anaesthesia. Most of them consist of a meningitis. We report a case of epidural abscess due to Staphylococcus aureus following spinal anaesthesia in a 62-year-old diabetic patient, diagnosed 45 days after the puncture with bacterial samples and magnetic resonance imaging. The pejorative neurological outcome required a laminectomy in spite of an efficient anti-staphylococcal treatment. PMID- 9750646 TI - [Sickle cell anemia and internal cerebral vein thrombosis]. AB - Cerebrovascular disorders are frequently associated with sickle cell disease, mainly in homozygous children. We report the case a 25-old-patient with known sickle cell disease who presented with coma inaugurated by manifestations of intracranial hypertension. CT revealed bilateral thalamic infarcts and angiography confirmed the thrombosis of internal cerebral veins. Treatment included heparin and blood transfusion. Severe cerebral oedema resulted in the lethal outcome three days later. PMID- 9750647 TI - [Cerebral fat embolism after closed leg injury]. AB - A 21-year-old man sustained a closed fracture of the leg from an industrial accident, without associated head trauma. The orthopaedic treatment consisted of immediate immobilization by setting leg in plaster. Two hours after admission, the Glasgow coma scale score was 10. Four hours after admission he developed a coma (Glasgow coma scale score = 7) with repetitive seizures. No lesion was visible on cerebral CT scan. Chest X-ray was unremarkable. Petechiae on the anterior chest wall and abdomen with bilateral mydriasis occurred. Thrombocytopenia with prothrombine time increase were observed. Magnetic resonance imaging, 27 hours after admission, showed high-intensity areas on T2 weighted views due to fat embolism. Retinal haemorrhages were observed. The bronchoalveolar lavage showing fat staining of tracheal aspirates confirmed the diagnosis of fat embolism. This case report emphasizes the possibility of predominant neurologic manifestations of a fat embolism and the diagnostic help of cerebral magnetic resonance imaging. PMID- 9750648 TI - [Dissecting ascending aortic aneurysm in pregnancy: simultaneous cesarean section and surgical aortic repair]. AB - The authors report a case of an ascending aorta dissection, occurring at 35 weeks of pregnancy. Emergency Caesarean section, and surgical aortic dissection repair under cardiopulmonary bypass, were undertaken. The aetiology, diagnosis and anaesthetic management are discussed. PMID- 9750649 TI - [Digestive hemorrhage disclosing an angiodysplasia and von Willebrand disease type 2]. AB - The authors report a case of a 76-year-old man, with not past history of abnormal bleeding, who suffered an acute, recurrent, intestinal haemorrhage from extensive angiodysplasia of the colon. Intestinal bleeding and angiodysplasia were associated with a von Willebrand's disease. Genetic analysis showed a point mutation in arginine 611 of the mature von Willebrand's factor subunit (A1 loop) confirming a von Willebrand's disease type 2. PMID- 9750650 TI - [Block of the lateral perforant branches of the subcostal and iliohypogastric nerves for proximal femur surgery]. AB - We describe an original method to block the lateral cutaneous rami of the subcostal and iliohypogastric nerves or'iliac crest point block'to complete plexular block of the lower limb for hip surgery. The local anaesthetic is injected in front of an osterofibrous orifice of the iliac crest. In nine cases out of ten, the lateral cutaneous rami of the iliohypogastric nerve pass through this orifice and in one case out of ten, it is the one arising from the subcostal nerve. This complementary block allows the surgical incision at the level of the great trochanter. PMID- 9750651 TI - [Locoregional anesthesia in the child. Expert Conference (Congress of the French Society of Anesthesia and Resuscitation, 1997)]. PMID- 9750652 TI - [Prehospital and early hospital management of a state of hemorrhagic shock of traumatic origin. 3rd Conference of experts in emergency medicine of the southeastern region]. PMID- 9750653 TI - [The 150th anniversary of chloroform. An anesthetic agent "yet more marvellous and terrible" than ether]. PMID- 9750654 TI - [Ketamine from an anticonvulsant perspective]. PMID- 9750655 TI - [Loading with hypertonic saline solution during pregnancy toxemia]. PMID- 9750656 TI - [Analgesia by continuous tibial nerve block after ischemia and amputation]. PMID- 9750657 TI - [Importance of bronchial block during peroperative rupture of the right bronchial trunk]. PMID- 9750658 TI - [Cerebral abscess caused by Nocardia]. PMID- 9750659 TI - [Locoregional anesthesia in children]. PMID- 9750660 TI - [Vascular filling in relative or absolute hypovolemia]. PMID- 9750662 TI - [How to prevent cauda equina syndromes occurring after continuous spina anesthesia?]. PMID- 9750663 TI - [The effect of tourniquet release time on analgesia effectiveness of the intra articular injection of morphine]. PMID- 9750664 TI - [Abdominal pain and laparoscopic surgery in children: mechanisms and treatments]. PMID- 9750665 TI - [Anesthetic management of a child with hydrocephaly or with a ventricular shunt]. PMID- 9750667 TI - [Approach to airway control in the 5 emergency services in Lyon]. PMID- 9750670 TI - [Preoperative evaluation of hemorrhagic risk]. AB - Evaluation of bleeding risk before surgery requires both precise knowledge of epidemiological data relating to haemostasis disorders and a true clinical approach to help guide the diagnosis. The patient's individual and familial history is recovered during both the interview with the anaesthesiologist and the clinical examination. Preoperative haemorrhage risk prevalence is 1/40,000 patients in patients with asymptomatic congenital haemostasis disorders with low bleeding and 1/2,000 patients for acquired asymptomatic haemostasis disorders. Deficits in haemostasis factors, i.e., congenital disorders with haemorrhage potential, have an overall prevalence of 1/6,500 patients. Whatever the clinical case, haemorrhage disorders will arise in patients with either congenital or acquired bleeding abnormalities without symptoms. To work in close collaboration with haemobiologists and to request appropriate biological screening tests, it is therefore important to take into account prevalence data according to the surgical environment from which the patient will benefit. PMID- 9750671 TI - [Preoperative evaluation of hemorrhagic risk]. AB - Should we consider that haemostasis tests may help guide the anaesthesiologist's therapeutic choices when the patients is on the operating table and bleeds without evident explanation? The response is yes, provided that the results will be available rapidly. Haemostasis tests that are performed in laboratory wards distant from the surgery room often prove to be unuseful in emergency situations as haemostasis abnormalities usually occur in a shorter time than is required to get biological test results. Devices located in the vicinity of the surgery room should therefore be available to the anaesthesiologist. Furthermore, these devices should be able to perform not only platelet counts and haematocrit determination, but other blood tests that may be useful to get further insights in the patient's haemostasis function. Such devices are already available, allowing rapid haemostasis tests in the surgery room and prompt decision with regard to appropriate transfusion policy. However, the haemostasis laboratory is still useful, as it permits to refine coagulation profile and also offers quality controls for those tests that were done in the surgery room. PMID- 9750672 TI - [Which perioperative therapeutic strategies in constitutional hemostasis disorders?]. AB - Type A and B haemophilias and von Willebrand's disease are the most common haemostasis disorders. Haemorrhage risks during both surgery and the time period surrounding surgery are often associated with these diseases. Consequently, normalization of the plasma level of the lacking protein and adapted therapeutic management will be the main objectives. Before undertaking any surgical procedure, the interview with the anaesthesiologist will allow detection of the disease. The type and the severity of the deficit at the origin of perioperative haemorrhage risks may be determined in collaboration with appropriate treatment centers. Therapeutic procedures (under the form of a protocol) will thus be defined. This multidisciplinary approach will make any surgical procedure feasible, whatever the type of haemostasis disorder. PMID- 9750673 TI - [How do we manage the hemorrhagic risk on hypovitaminosis K and treatments with antivitamin K]. AB - Vitamin K deficiency leads to a deficit in vitamin K-dependent factors, resulting in either hypocoagulability and a decrease in the Quick one-stage prothrombin time expressed as the prothrombin time (PT), or in an increase in INR in patients receiving oral anticoagulation. The anesthesiologist's objective is to bring these values back into the safety range before surgery, i.e., above 50% for PT and below 1.5 for INR. The method to be used will be chosen according to the urgency of the correction. Safety ranges may be reached in 6-12 h following oral or parenteral administration of vitamin K. A 5-mg dose is usually sufficient. If the deficit in vitamin K-dependent factors requires immediate correction, intravenous administration of PPSB should be done. The minimum time during which antivitamin K treatment may be disrupted after surgery depends on both the possibility of restarting oral treatment and the risk of postoperative haemorrhage. During this period, the need for an anticoagulation treatment using heparin should be discussed according to the risk of thrombosis. PMID- 9750674 TI - [The paradox of disseminated intravascular coagulation]. AB - Disseminated intravascular coagulation (DIC) is an abnormal extension of a coagulation process normally limited in time and space. DIC results from the release of free thrombin in circulating blood, as thrombin synthesis and inactivation mechanisms non longer function properly. In addition to microthrombosis, acute DIC may lead to haemorrhage related to activation of the coagulation process. According to the disease aetiology, emergency treatment may become a top priority, in case of haemorrhage, shortage of platelets and coagulation factors related to increased use of these blood components requires replacement treatment with infusion of platelets and fresh frozen plasma. As treatment of the procoagulation process, heparin should not be recommanded in acute DIC with haemorrhage. Based on rational theoretical data, current evaluation of antithrombin therapy is in progress. PMID- 9750675 TI - [The good use of drugs derived from blood]. AB - Rational use of blood-derived products should only be viewed in close collaboration with haemostasis laboratory teams, pharmacists and transfusion centers. The crucial role of a detailed clinical interview can never be emphasized enough. The use of blood-derived products or the elaboration of new therapeutic strategies should always be mentioned in a written protocol. These procedures clearly belong to quality control implementation. The patient as well as the surgical team should always be aware of haemostasis procedures. PMID- 9750676 TI - [The conformity of anesthesia materials with regulations: what about the reserve oxygen cylinder?]. PMID- 9750677 TI - [Dobutamine echocardiography for the preoperative evaluation of patients for surgery of the abdominal aorta]. AB - OBJECTIVE: The aim of the study was to assess the value of dobutamine echocardiography (DE) for detecting coronary artery disease (CAD) in patients scheduled for abdominal aortic surgery. STUDY DESIGN: Preliminary prospective open study. PATIENTS: Thirty-three consecutive patients due to undergo effective abdominal aortic surgery, assessed by preoperative DE. METHODS: Previous myocardial infarction and atherosclerotic risk factors (RF) were noted. Incremental doses of dobutamine were administered in order to reach 85% of age predicted maximal heart rate. The occurrence of regional wall motion abnormalities was considered as a positive test. In this case a coronary angiography was performed. RESULTS: Four patients had a history of angina pectoris. DE was not interpretable in five patients. Among the patients without symptoms, 12 had three RF or more, 12 had less than three RF. In eight patients with a positive test, coronary angiography showed one or more significant main coronary artery stenoses. All patients with angina pectoris had a positive test. None of patients without symptoms and less than three RF had a positive test, one third of patients with no symptomatology but with three RF or more had a positive test (P < 0.05). CONCLUSION: DE has the ability to identify patients with asymptomatic CAD. DE is recommended in patients with high probability of CAD, i.e. with three RF or more. PMID- 9750678 TI - [Controlled analgesia in a burn patient: fentanyl sparing effect of clonidine]. AB - OBJECTIVE: To determine the sparing effect of clonidine (C) on fentanyl (F) demand in burned patients under PCA. STUDY DESIGN: Prospective, randomized, double blind study versus placebo. PATIENTS: Twelve consecutive patients with mean burn surface area of 20 +/- 9%, studied between the third and the eighth day post-burn. METHODS: F was delivered by a PCA pump (bolus: 1 mg.kg-1). In the morning, burn patients received additional F (5 mg.kg-1) before hydrotherapy. After randomisation, C or placebo (P) were alternatively infused over 24 hours. Demands for F during the morning, the afternoon and the evening were noted. Pain scores were measured on a visual analogic scale. In eight patients, plasma levels of F (pF) were iteratively measured. Heart rate, arterial pressure, respiratory rate and SpO2 were monitored. RESULTS: Analgesic demands were 19.5/day under P and 9.5 under C (P < 0.0001). Pain reoccurred for pF of 4.1 under C vs 5.7 under P (P < 0.05) with same pain scores in the two groups. After a pain stimulus, pain scores were lower under F, despite lower pF (P < 0.05). Arterial pressure and heart rate were significantly lowered during the first hour of C infusion. CONCLUSION: Doses of F and pF required to reach analgesia were very high. Adding C decreases by 50% the F demand and lowers pF. Minor haemodynamic effects were observed during the first hour of C infusion. PMID- 9750680 TI - [Pressure exerted on tracheal cuffs in intubation in the presence of nitrous oxide]. AB - OBJECTIVE: To compare the time course of endotracheal tube cuff pressures in presence of nitrous oxide (N2O), obtained in a tracheal model with those measured during clinical anaesthesia. STUDY DESIGN: Experimental and clinical prospective study. MATERIAL: Twelve brands of low-pressure tracheal tubes. METHODS: The pressure changes in the cuffs were measured over a three-hour-period in presence of a N2O (50 vol%)/O2 (50 vol%) mixture and mechanical ventilation, the tube being inserted either in a tracheal model or in the trachea of patients during general anaesthesia. RESULTS: The results obtained in vitro were correlated with those measured in the patients. Therefore the tracheal model is a helpful guide for the choice of endotracheal tubes. PMID- 9750679 TI - [The good use of antibiotics in intensive care: results of a program for rationalization of prescriptions]. AB - OBJECTIVE: To assess the impact of an antibiotic prescribing programme in a intensive therapy unit. TYPE OF STUDY: Prospective comparative study. METHODS: We compared antibiotic prescriptions and bacterial susceptibility to antimicrobial agents before and after introduction of a programme focusing on injection control and therapeutic indications. RESULTS: The introduction of the programme resulted in a major decrease in antibiotic administration. Moreover, the susceptibility of Pseudomonas aeruginosa to ticarcillin increased from 40 to 68%, and susceptibility of Staphylococcus aureus to methicillin increased from 55 to 73%. CONCLUSIONS: Antibiotic control policies must be considered integral to any effort to decrease resistance and cost of therapy with antibiotics. PMID- 9750681 TI - [Prolonged curarization with suxamethonium in a four-week old infant]. AB - A case of a 28-day-old infant who developed suxamethonium apnoea is described. He was found to be homozygous for atypical cholinesterase. Main characteristics of this disorder are reviewed. Other causes of prolonged apnoea in infants recovering from anaesthesia for surgery of pyloric stenosis are discussed. PMID- 9750682 TI - [Acute cardiogenic postoperative edema after doxorubicin (adriamycin) chemotherapy]. AB - We describe a case of postoperative congestive heart failure in a young woman of physical class ASA 1, following breast cancer surgery. Preoperatively she had been treated with doxorubicin (Adriamycin) 450 mg.m-2, total dose, associated with breast and ovarian radiotherapy. This association was probably the cause of postoperative heart failure. Twenty-four hours after surgery, a two-dimensional echocardiography showed a severe left ventricular dysfunction, whereas preoperative clinical assessment was unremarkable. Doxorubicin cardiotoxicity can be acute, subacute and delayed as in our case. Clinical assessment and ECG are not sensitive indicators of such cardiac damage. Preoperative echography is the technique of choice for the evaluation of the cardiac status of a patient treated with a high cumulative dose of doxorubicin and mediastinal irradiation. PMID- 9750683 TI - [Implanted automatic defibrillator after ventricular fibrillation treated with semi-automatic defibrillation]. AB - We report two cases of out-of-hospital ventricular fibrillation treated without delay, with basic life support practiced by the witness, followed by a successful defibrillation by paramedics with a semi-automatic defibrillator. In the subsequent month, a cardioverter-defibrillator was implanted. In one patient, a ventricular tachycardia occurring 10 months later and a ventricular fibrillation 9 months later in the other respectively, were successfully reversed by the implanted defibrillator. These two cases illustrate the value of the "survival chain" concept (undelayed alert, basic life support by witness, early defibrillation by paramedics with a semi-automatic defibrillator, advanced life support by a physician) as well as the benefit of the implanted cardioverter defibrillator. PMID- 9750684 TI - [Central venous catheterization...don't forget the caval filter]. AB - Among the complications of central venous line insertion, entrapment of guidewire by inferior vena cava filter has been exceptionally reported. Usually the disengagement attempts resulted in a filter migration. We report a case of guidewire entrapment successfully treated with interventional radiology techniques. PMID- 9750685 TI - [Secondary cubital paralysis from prolonged thoracic drainage]. AB - In intensive therapy patients, thoracic drains are usually inserted in the lateral part of thorax with the extension tube crossing the posterior aspect of the upper limb. We report the cases of two sedated patients who experienced ulnar palsy from a thoracic drain located behind their elbow. PMID- 9750686 TI - [Rupture of the intra-abdominal spleen as a cause of hemothorax]. AB - We report a case of a 17-year-old patient with an haemothorax related to an intra abdominal spleen rupture. The nature of this haemothorax is unusual and the problem is how to obtain the etiology in this case. The spleen was in intra abdominal position on all radiological examinations and was the source of bleeding. Therefore, laparotomy is the only convenient therapy. PMID- 9750687 TI - [Recurrent hematomas and normal standard hemostasis tests]. AB - The authors report the case of a 22-month-old boy experiencing a voluminous subcutaneous haematoma, 72 hours after a head trauma. Two subsequent drainages of this haematoma were required because of its recurrence. The child, whose parents had blood relations, suffered from recurrent bleeding since his birth. A standard haemostasis assessment including prothrombin time, activated partial thrombopiastin time, bleeding time, concentration of fibrinogen and platelet count was unremarkable. Therefore, coagulation factors were explored. An inherited factor XIII deficiency (less than 2%) was recognized. A new drain was inserted, after administration of factor XIII concentrate. The time course of the haematoma was favourable. After discharge, the prophylactic therapy consisted of an injection of factor XIII concentrate (50 Ul.kg-1) every 5 weeks. PMID- 9750688 TI - [A cause of intubation failure: a subglottic bridge]. AB - We report the case of a patient with a history of facio-thoracic burns, the treatment of which included prolonged intubation, whose trachea could not be intubated because of a subglottic obstacle. The ventilation was easily controlled with a laryngeal mask. At the end of surgery for postburn cheloids, laryngoscopy through the laryngeal mask showed a transversal subglottic laryngeal band, a probable sequela of the previous prolonged intubation. The band was resected one week later. The conventional indicators for difficult intubation cannot detect the laryngotracheal obstacles to tracheal tube insertion. PMID- 9750689 TI - [Improvement of glottis visualization with a McCoy blade]. AB - Difficult intubation remains one of the major risks in anaesthetic practice. Recently, as other anaesthetic societies, the French Society for Anaesthesia and Intensive Care (SFAR) has produced algorithms for the management of a difficult intubation. New laryngoscopes and blades have been marketed in recent years, however their place in these algorithms remains unclear. In this preliminary study, we compared the laryngoscopic view in 100 consecutive patients during tracheal intubation with the McCoy laryngoscope first in its normal position and after levering the distal part of the blade. All patients were included in Mallampati classes 1 and 2. Among them, 16% were classified as Cormack and Lehane grades 3 or 4 when using the blade in the normal position. These data confirm previous observations showing that the McCoy blade in normal position performs poorly as compared with the Macintosh blade. Conversely the levering of the distal part of the blade significantly decreased the incidence of Cormack and Lehane grades of 3 or 4 (2 versus 16%, P = 0.001). It is concluded that the McCoy blade is not convenient for its routine use in patients not to be at preoperatively known risk of difficult intubation. This blade significantly improves intubating conditions. Defining the exact place of this new blade in difficult intubation algorithms requires further studies. PMID- 9750690 TI - [Hydroxethyl starch: effects on hemostasis]. AB - HES are high-polymeric glucose compounds obtained via hydrolysis and subsequent hydroxyethylation from the highly-branched amylopectin contained in maize. Initially, the HES were only characterized by their in vitro molecular weight (Mw), without consideration of the in vivo hydrolysis by alpha-amylase. The degree of substitution and the molar substitution ratio quantify the hydroxyethylation. The glucose units can be substituted at carbon 2, 3 and 6 leading to various substitution patterns. This pattern is described with the C2/C6 hydroxyethylation ratio. The higher the degree of substitution and the C2/C6 ratio, the less the starch is metabolized. The in vitro Mw, the degree of substitution and the C2/C6 ratio are the main determinants of the in vivo Mw which is clinically relevant. Haemorrhagic complications that occur after infusing larger volumes of HES can be avoided with a starch of low in vivo Mw. This is not only due to a lesser effect on the coagulation system which prevents an acquired type I von Willebrand syndrome, but also to a smaller decrease in platelet volume, since platelet volume and platelet function are positively correlated. In addition, HES with low in vivo Mw has significantly better rheological effects than HES with a high in vivo Mw, as high Mw macromolecules affect plasma viscosity negatively. Furthermore high Mw HES macromolecules lead to a distinctive decrease in fibronectin concentration that reflects saturation of the reticuloendothelial system. Another advantage of low in vivo Mw HES is its rather short half-life. Patients with an increased bleeding risk, microcirculatory disturbance or affected RES should receive HES with low in vivo Mw. In the future, HES should be mainly characterized by the in vivo and not the in vitro Mw. PMID- 9750691 TI - [Autotransfusion of blood salvaged in a surgical emergency department]. PMID- 9750692 TI - [Danaparoid (orgaran) for continuous venovenous hemodiafiltration in a patient with heparin-induced thrombocytopenia]. PMID- 9750693 TI - [Hypercapnia in pulmonary embolism]. PMID- 9750694 TI - [Neuroleptic malignant syndrome induced by risperidone]. PMID- 9750695 TI - [Acute intoxication with disopyramide]. PMID- 9750696 TI - [Percutaneous dilatation tracheotomy a useful technique for emergency technique?]. PMID- 9750697 TI - [Percutaneous dilatation tracheotomy: the technique of Griggs or Ciaglia? A response]. PMID- 9750698 TI - [Allergo-anesthesia consultation: not enough patients are tested after an anaphylactoid anesthetic incident]. PMID- 9750699 TI - [The concept of AIVOC]. PMID- 9750700 TI - [The use of perfusion apparatus for objective concentration of intravenous anesthesia]. PMID- 9750701 TI - [Pump destination in AIVOC: procedures for receiving, utilization and maintenance]. PMID- 9750702 TI - [Pre-emptory syringes of Diprivan: regulatory, practical and economic aspects]. PMID- 9750703 TI - [Materiopharmacovigilance: application to a couple of products plus application to AVIOC]. PMID- 9750704 TI - [The position of the neurosurgical patient. Artificial feeding in cranial trauma]. PMID- 9750705 TI - [Comparative hemodynamic effects of sevoflurane and halothane at tele-expiratory concentration in intubation of infants]. AB - OBJECTIVE: To compare the cardiovascular changes at the end-tidal concentrations of sevoflurane versus halothane required for tracheal intubation in infants (intubation MAC). STUDY DESIGN: Prospective randomized study. PATIENTS: Thirty two infants, ASA physical status 1 or 2, scheduled for elective surgery, randomized to receive either halothane or sevoflurane for anaesthetic induction by inhalation. METHODS: Cardiovascular and echocardiographic data were recorded in both groups at baseline, and at the end-tidal concentrations needed for intubation. RESULTS: Intubation MAC was significantly less with sevoflurane than with halothane in infants. Sevoflurane did not change heart rate (HR) and cardiac index (CI) compared to values when awake. Sevoflurane significantly decreased blood pressure, systemic vascular resistance (SVR) and shortening fraction (SF). Myocardial contractility assessed by stress-velocity index (SVI) and stress shortening index (SSI) decreased significantly at the intubation MAC, but did not fall into the abnormal range. Halothane caused a greater decrease in HR, SF, SSI, and CI than sevoflurane. CONCLUSIONS: Sevoflurane decreases cardiac output less than halothane in infants at the intubation MAC, due to a lower end-tidal concentration at intubation MAC and to less effects on haemodynamic variables. PMID- 9750706 TI - [Medical treatment of spinal cord injury in the acute stage]. AB - OBJECTIVES: To evaluate the effect on neurologic outcome and the safety of nimodipine (N), methylprednisolone (M), or both (MN) versus no medical treatment (P) in spinal cord injury at the acute phase. STUDY DESIGN: Prospective, randomized clinical trial. PATIENTS: One hundred and six patients with a spinal trauma, including 48 with paraplegia and 58 with tetraplegia. METHOD: After eligibility, patients were randomly allocated in one of the following groups: M = methylprednisolone 30 mg.kg-1 over 1 hour, followed by 5.4 mg.kg-1.h-1 for 23 hours, N = nimodipine 0.015 mg.kg-1.h-1 over 2 hours followed by 0.03 mg.kg-1.h-1 for 7 days, MN or P. Neurologic assessment (ASIA score) was performed by a senior neurologist before treatment and at the 1-year follow-up. Early spinal decompression and stabilization was performed as soon as possible after injury. RESULTS: One hundred patients were reassessed at the 1-year follow-up. Neurologic improvement was seen in each group (P < 0.0001), however no neurologic benefit from treatment was observed. Infectious complications occurred more often in patients treated with M. Early surgery (49 patients), within the first 8 hours did not influence the neurologic outcome. The only predictor of the latter was the extent of the spinal injury (complete or incomplete lesion). CONCLUSION: Currently, no evidence of the benefit of medical treatment in this indication is existing. Because of the lack of clinical studies proving efficacy of pharmacological treatment in this specific pathology, a systematic use of medications cannot be recommended. PMID- 9750707 TI - [Bronchial lobar obstruction from a sucralfate tablet: a rare cause of acute respiratory insufficiency]. AB - We report two cases of patients with chronic respiratory disease who experienced an asphyxia after aspirating a sucralfate tablet that occluded a lobar bronchus. In adults, a foreign body is a rare cause of acute respiratory failure from tracheobronchial occlusion. The sucralfate tablet has the physical property of expanding rapidly when wet (contact with mucosa). After aspiration, the tablet expands to a larger size and can occlude a lobar bronchus, causing acute respiratory failure. In patients at risk of aspiration, we recommend the use of sucralfate in liquid or suspension form. PMID- 9750708 TI - [Post-transfusion purpura: and cause of severe postoperative thrombopenia]. AB - A 59-year-old woman developed an acute and severe thrombocytopenia (platelet count below 10.10(9).L-1) with active bleeding, 6 days after a massive transfusion for intraoperative haemorrhagic shock. The diagnosis of post transfusion purpura (PTP) was confirmed by the presence of an allo-antibody directed against HPA-1a platelet antigen. The patient and her daughter had a rare HPA-1b platelet phenotype, but also belonged to the HLA DR3 phenotype, frequently associated with PTP. This case shows the therapeutic difficulties of postoperative PTP. Despite active bleeding, this syndrome requires the discontinuation of transfusions of incompatible platelets. Transfusion of phenotyped platelets is often inefficient. Red cell concentrates must be platelet and plasma free. Immunomodulating therapy can shorten the time course. Preventive measures, particularly autologous transfusions, are necessary for subsequent haemorrhagic surgery or parturition. PMID- 9750709 TI - [Four hours for a record, or a severe fuminating cellulitis: can Saccharomyces cerevisiae be the causal agent?]. AB - A 31-year-old woman presented with a subcutaneous cellulitis which occurred within four hours following a minor wound of a knee. This very short delay could be explained neither by the health state, nor the mechanism of injury, nor the bacteria usually responsible for such a cellulitis. Considering the clinical characteristics (high gas production) and the professional context (wine cellar employee), Saccharomyces cerevisiae, a yeast used for wine or bread production, may explain this complication. PMID- 9750710 TI - [Blood volume changes caused by position under general anesthesia]. AB - The cardiovascular changes in response to general anaesthesia are related to several interacting mechanisms. In addition to the intrinsic effects of anaesthetic agents on the heart and vessels which depress the physiological mechanisms of adaptation, the additive independent effects of posture and mechanical ventilation on intravascular blood volume have to be considered. Physiopathologic studies show that the ultimate related mechanism is the decrease of venous return to the heart. The major effect of posture is the change in the distribution of the blood volume, as the posture modifies the influence of gravity in addition to direct vascular compression and stretching. The deterioration of the cardiac venous return results in dramatic or insidious clinical consequences which lead finally to a low cardiac output. As cardiac function is not a limiting factor of output, any decrease of blood pressure in a patient with a healthy heart must be considered as an hypovolaemic state due to an abnormal contents to container ratio, and must be managed as such. PMID- 9750711 TI - [Monitoring and maintenance of blood volume in intracranial surgery]. AB - Intraoperative monitoring usually does not assess intravascular blood volume of the patient. The variations of the systolic blood pressure and the right auricular pressure allow to appreciate the intravascular blood volume changes and their consequences on the cerebral perfusion pressure. However, when the autoregulation of the cerebral blood flow is strongly depressed, the evaluation of the cerebral perfusion pressure alone is inadequate. In this case, the optimal monitoring should be metabolic (jugular bulb venous oxygen saturation) to finally assess the cerebral flow-metabolic coupling. PMID- 9750712 TI - [The cerebral venous outflow tract]. AB - It is generally assumed that the jugular veins are the only significant path for cerebral venous drainage. Little attention has been given to the possible role of other venous paths for the cerebral venous outflow. In fact, the meningorachidian venous plexus acts as the major outflow tract of cerebral circulation in the upright position. This phenomenon is linked with postural variations in cerebrospinal fluid pressure. It is difficult to assess the significance of this posterior venous plexus under physiopathologic conditions, which probably depends on the extent of the anastomoses between the two systems. PMID- 9750713 TI - [The effect of position on intracranial pressure]. AB - The question as to whether the head and trunk of neurosurgery patients should be elevated remains controversial. This question is particularly important when intracranial hypertension is present. Head up position may have beneficial effects on intracranial pressure (ICP) via changes in mean arterial pressure (MAP), airway pressure, central venous pressure and cerebro spinal fluid displacement. However, in some circumstances, head up position may decrease MAP which in turn will result in a paradoxical rise in ICP through autoregulation mechanisms. Therefore, the degree of head elevation has to be titrated by evaluating the most adequate cerebral perfusion pressure (CPP) for each patient by means of transcranial Doppler or measurement of jugular venous blood oxygen saturation. Head elevation above 30 degrees should be avoided in all cases. In most patients with intracranial hypertension, head and trunk elevation up to 30 degrees is useful in helping to decrease ICP, providing that a safe CPP of at least 70 mmHg or even 80 mmHg is maintained. Patients in poor haemodynamic conditions are best nursed flat. CPP is thus the most important factor in assessment and monitoring when considering head elevation in patients with increased ICP. PMID- 9750714 TI - [Cerebral risks in surgical position: position of the body, position of the brain]. AB - The posture of the neurosurgical patient depends first on the approach and working position of the surgeon, and the spatial orientation during surgery. Venous outflow is probably the major factor for optimal brain retraction and for preventing venous haemorrhage. No one of the postures presently in use is fully free from the risk of air embolism. Respiratory and cardiac diseases are often limiting factors for the choice of a posture. Thoracic and abdominal compression must be cautiously avoided. PMID- 9750715 TI - [The sitting position in neurosurgery: the viewpoint of the surgeon]. AB - The sitting position offers the benefits of better access to the apex of the posterior fossa, and an improved exploration and dissection because blood and cerebral spinal fluid drain away from the operative site. Specific complications of the sitting position include cardiovascular instability, jugular venous obstruction, airway oedema, quadriplegia, displacements of catheters and the endotracheal tube, ulnar, sciatic and lateral peroneal nerve compression, venous air embolism, and tension pneumocephalus. In the only existing comparative study, the differences were an increased bleeding in the horizontal position and a better cranial nerve preservation in the sitting position. This argues strongly for teaching and use of the sitting position whenever surgically indicated. PMID- 9750716 TI - [The sitting position in neurosurgery: the viewpoint of the anesthetist]. AB - The sitting position can be used safely in neurosurgery, if meticulous attention is given to the positioning and the monitoring of the patient during surgery. Venous air embolism remains the most frequent complication. Hypotension is the second complication. A rigorous patient selection of patients, the experience of the anesthesiologist and the neurosurgeon remain the main factors for the choice of the sitting position. PMID- 9750717 TI - [Surgery of the supratentorial and ventral spine: view-point of the surgeon]. AB - The prone position for posterior fossa and cranio-spinal surgery is becoming the standard posture for neurosurgical teams. We analyze the reasons of such a change and specify indications of this posture. Thereafter, the procedure for optimal positioning is considered and the benefits of the prone position for the neurosurgeon and the anaesthetist are discussed. The selected references originate from Medline search (1991-1997), the Year Book of Neurology and Neurosurgery and the major neurosurgical review articles published between 1980 and 1996. PMID- 9750718 TI - [Perioperative effects of the prone position: anesthesiologic aspects]. AB - The prone position is commonly used for surgery of the spine and the posterior fossa, and is well tolerated by the majority of patients. As long as the abdomen is not compressed, the physiologic impact of this position on cardiorespiratory function is minor, in some cases even less than with the supine position. However extremes of position, particularly of the head and neck, are poorly tolerated and may lead to a variety of severe neurological complications. In addition, several specific forms of pre-existing pathology may predispose the prone patient to major cardiorespiratory complications. In this paper we have systematically reviewed the English and French literature from 1991 to 1997 using Medline Search of peer reviewed journals for the search terms "prone position" and "prone position and venous air embolism". The 330 collected references were reviewed for quality. In combination with review of current standard textbooks these references form the basis for the current report. PMID- 9750719 TI - [Intracranial surgery in the lateral position]. AB - The lateral posture (Park Bench) is now widely used. It provides a comfortable position for the surgeon and a convenient brain relaxation. However the positioning is complex and carries a risk of stretching of the brachial plexus. Spatial orientation is more difficult and requires surgical experience. The lateral position does not modify the haemodynamic and respiratory function in healthy patients. In the opposite, various cardiac or respiratory effects of right or left lateral position can occur in patients with cardiac and/or respiratory pathology. PMID- 9750720 TI - [Catabolic aspects of cranial trauma]. AB - During their stay in the intensive care unit, head-trauma patients develop a hypermetabolic and a hypercatabolic status. Their nitrogen balance is highly negative and the muscular proteolysis is largely increased. The nitrogen losses originate mainly in muscles, resulting in muscle wasting and weakness. The whole protein synthesis remains quite normal, but this does not reflect the reality, as muscular protein synthesis is decreased, while hepatic protein synthesis is increased. The increased proteolysis seems to be due to the activation of the three proteolytic pathways, particularly the ATP-ubiquitin-dependent pathway. The causes of the increased muscle proteolysis in head trauma patients remain unclear. The increased glucocorticoid release, which is integrated in the acute phase response to injury, could be one of them. Glutamin, vitamin or zinc supplementation has been proposed in head trauma patients. The use of glucocorticoid antagonists, recombinant growth hormone or anti-cytokines are our fields of research. PMID- 9750721 TI - [Nutritional aspects of cranial trauma]. AB - In head-injured patients the nutritional support is aimed to prevent denutrition status usually observed. The adequate amount of calories depends on the basal metabolism (as calculated with the Harris Benedict equation). It has to be increased in case of fever (by a 0.1 factor per degree above 37 degrees C), sepsis (by a 0.1 to 0.2 factor) or when sedation is discontinued (by a 0.3 factor). The increased proteolysis is not modified by the associated treatment and results in an inevitable protein loss, whatever the qualitative change in nutritional support. In clinical practice, the nutritional support has to be adjusted continuously to the needs of the patient, to avoid a more pronounced denutrition due to the summation of daily nutritional deficits. PMID- 9750722 TI - [Early enteral feeding in cranial trauma]. AB - Until some years ago, patients suffering from head injury were poorly fed and nutrition was not a primary concern in the medical treatment of these patients. To date, six studies on head-injury patients have examined the effect of nutritional support on their outcome. All showed that lack of adequate nutrition contributed to increased mortality and morbidity. Head-injured patients on conventional enteral nutrition receive significantly less calories and proteins, resulting in an increased morbidity and mortality rate. Most of the severely head injured patients receiving enteral nutrition do not tolerate enteral feedings because of abnormal gastric emptying. The mechanisms of altered gastric function remain obscure. Increased intracranial pressure, cytokines, corticotropin releasing factor, opiates, and other agents may all play a role. Impaired gastric motility has led to increasing use of small bowel feeding in head-injured patients. Jejunal feeding enables a higher caloric input and a better nitrogen balance. Moreover, it enables early enteral administration of nutrients in a safe and efficient way. Early administration of nutrients may be extremely important as it seems to decrease the hypermetabolic response to traumatic injury. Therefore, early jejunal enteral feeding may become an important cornerstone in the medical management of head-injured patients. PMID- 9750723 TI - [Streptococcus B meningitis after peridural obstetric anesthesia]. PMID- 9750724 TI - [Cortical blindness in pregnancy toxemia. Transient amaurosis from a decrease of cerebral perfusion pressure]. PMID- 9750725 TI - [Profound normovolemic hemodilution in a pediatric Jehovah's Witness patient and organ donation: what are the limitations?]. PMID- 9750726 TI - ["Blockade" for leg fracture repair of the upper humerus]. PMID- 9750727 TI - [Effectiveness of peripherally administered opioids for spinal or peripheral block]. PMID- 9750728 TI - [What are the effects of peripherally applied clonidine?]. PMID- 9750729 TI - [Practical aspects of use of nitrous oxide in children]. PMID- 9750730 TI - [Use of anesthetic and sedative agents in a subject treated with a monoamine oxidase inhibitor]. PMID- 9750731 TI - -The clinical use of the cuffed oropharyngeal airway (COPA)-. AB - OBJECTIVE: To assess the performance of the COPA device during general anaesthesia. STUDY DESIGN: Prospective, clinical, open study. PATIENTS: Eighty patients scheduled for short elective surgical procedures under general anaesthesia not requiring tracheal intubation. METHOD: After premedication (midazolam, atropine), anaesthesia was induced with propofol (154 +/- 40 mg = 2.47 +/- 0.8 mg.kg-1) and alfentanil (1.14 +/- 0.43 mg). The COPA device was inserted in a fashion similar to a Guedel airway device. The device was evaluated on the following criteria: correct choice of COPA size, ease of insertion, ability to obtain or maintain patent airway. Adverse reactions were noted, such as coughing, nausea, regurgitation, inhalation, and sore throat. The overall rating of the COPA as a "hand free device" was evaluated on the basis of excellent, good, fair, and poor. RESULTS: Insertion of the device was easy and in 70 cases successful on the first attempt. Jaw thrust on head tilt was necessary in half the cases. No patient necessitated intubation because of hypoxaemia or airway obstruction. Adverse reactions occurred in few cases and consisted of sore throat (always moderate) in 10% of the cases. COPA was evaluated as excellent or good in 80% of the cases. CONCLUSION: COPA is a convenient device for airway management in fasting patients undergoing general anaesthesia for elective surgery in the supine position, in whom tracheal intubation is not indicated. PMID- 9750732 TI - -Anesthesia and analgesia practice patterns in French obstetrical patients-. AB - OBJECTIVE: To assess the rate of epidural analgesia (EA) for parturition and the techniques of anaesthesia for Caesarean section (CS). STUDY DESIGN: Retrospective study. PATIENTS: A series of 84,235 deliveries. METHODS: The series was extracted from a total of 770,054 deliveries carried out in 1991, according to the number of births in each hospital (1/1 if the births were < or = 100 per year, up to 1/25 if they were > or = 2,000 per year). The data analyzed included: anaesthesia technique, whether or not there was an anaesthetist on night duty at the hospital, birth rate in the hospital, type of hospital: university (UH), general (GH) or private (PH). For vaginal deliveries, the mode of labour commencement (spontaneous or induced), the multiplicity of pregnancies and a history of past CS were also noted. RESULTS: Vaginal deliveries: the overall rate of EA was 37.2%. EA were not carried out in 5% of maternity hospitals. In cases of spontaneous labour, the average rate was 32.1%, significantly less than for induced labour (59.6%, P < 0.0001) and in cases of previous CS (39%, P < 0.05). There was no statistical difference in cases of multiple pregnancies (35.7%). The average rate of EA was correlated to the number of annual births (P < 0.001) and was increased when the anaesthetist was present in hospital at night (P < 0.001). It was also significantly lower in GH (P < 0.001) than in UH or PH, which were equivalent. Scheduled CS: general anaesthesia (GA) was carried out at a significantly higher rate than regional anaesthesia (RA) (49.7% vs 48.4%, P < 0.05). In 15.1% of hospitals, RA was not available. The incidence of RA was influenced neither by the rate of annual births nor by the presence of the anaesthetist in the hospital during night. However, RA was significantly less frequent in GH (46.3%, P < 0.001) than in UH (48.6%) and in PH (53.6%) which were equivalent. CS during labour: the incidence of RA was significantly higher than GA (53.2% vs 44.1%, P < 0.001). In 17.1% of hospitals, RA was never carried out. The rate of RA was correlated to the size of the maternity hospital, and significantly higher (P < 0.001) when the anaesthetist was present in hospital during night. The differences between UH, GH and EP were the same than for scheduled CS. CONCLUSION: In France in 1991, the average rate of 37.2% for EA for obstetrics was high when compared to the rate in United Kingdom. It was equivalent to those in United States and Ontario, Canada. The discrepancies between hospitals were mainly related to structural and organizational factors. The influence of the size of the maternity hospital, the 24-hour service of EA was also shown in other studies. However, the difference between GA and UH and PH is a French particularity. The high rate of GA for CS differs largely with those in the UK or the USA. The time saving aspect of GA was probably an important factor for the choice of this technique. This study must be reactualized and enlarged to determine the demand of EA for labour by parturients and obstetricians. PMID- 9750733 TI - -The effect of ondansetron on intracranial pressure and cerebral perfusion pressure in neurosurgical patients-. AB - OBJECTIVE: To determine the effect of ondansetron on intracranial pressure (ICP), mean arterial pressure (MAP) and cerebral perfusion pressure (CPP). STUDY DESIGN: Prospective, comparative, randomized double-blind study. PATIENTS: Twenty-six patients undergoing intracranial surgery. METHOD: Induction was obtained with propofol (1-2.5 mg.kg-1), fentanyl (1.5 micrograms.kg-1) and pancuronium (0.1 mg.kg-1), and maintenance was achieved with propofol and fentanyl. Intermittent positive pressure ventilation was used to ensure mild hypocapnia at 35 +/- 2 mmHg. Positioning of the patient was followed by 15 minutes steady-state. Patient received thereafter either 8 mg ondansetron or a placebo intravenously. The ICP was measured using a lumbar malleable spinal needle. CPP was calculated using the formula CCP = MAP-ICP. All variables were measured every minute for 15 minutes. RESULTS: The ICP, MAP and CPP did not differ between the two groups. There were no differences in the highest ICP values in patients receiving either ondansetron or placebo (11 +/- 5 versus 9 +/- 5, mean +/- SD), respectively. CONCLUSION: Intravenous administration of 8 mg ondansetron affects neither cerebral hemodynamics nor ICP. PMID- 9750734 TI - -The effect of desflurane on cerebral blood flow velocity and cerebrovascular reactivity to CO2 in children-. AB - OBJECTIVE: To assess in children with a transcranial Doppler the effect on cerebral blood flow velocities of desflurane, whose cerebral vasodilator effects have been studied in animals and in adults with intracranial lesions. STUDY DESIGN: Prospective clinical study. PATIENTS: Ten healthy children, mean age: 3.4 yr, ASA physical class 1, undergoing minor urologic surgery, were included in this study. METHOD: Induction was obtained with atropine 10 micrograms.kg-1, fentanyl 3 micrograms.kg-1 and propofol 3 mg.kg-1. Endotracheal intubation was facilitated by atracurium 0.3 mg.kg-1. Mechanical ventilation, with a 50% air/oxygen mixture was adjusted to achieve an end-tidal CO2 (PETCO2) level of 38 +/- 2 mmHg. Monitoring included measurement of mean arterial blood pressure (MAP), heart rate, PETCO2, SpO2 and end-tidal desflurane concentrations (FETDes). Mean blood flow velocities (Vmean) were measured in the middle cerebral artery using a bi-directional 2 MHz TCD system (EME-TC 2000 S). A first TCD measurement followed intubation (T1). Thereafter, desflurane was adjusted to 1 MAC. Six other TCDs were recorded each minute until FETDes reached the inspired fraction (T2 T7). Thereafter, CO2 reactivity was assessed with a hypocapnia test, induced by hyperventilation. Measures were done at T8 (PETCO2: 33 +/- 1 mmHg), T9 (PETCO2: 29 +/- 1 mmHg), and T10 (initial PETCO2: 38 +/- 1 mmHg). All these measurements were made before starting surgery. Analysis of variance (ANOVA) was used to analyse the data (P < 0.05 was considered as significant). RESULTS: The Vmean and heart rate increased significantly with increasing concentrations of desflurane (Vmean from 68 +/- 27 to 106 +/- 30 cm.s-1 and heart rate from 109 +/- 17 to 136 +/- 15 b.min-1 between T1 and T7). During hypocapnia, Vmean decreased to 68 +/- 23 cm.s-1 at T9, and returned to normal values with PETCO2 at 38 mmHg at T10. SpO2 remained unchanged. Mean arterial pressure was stable from T1 to T7, but decreased significantly at T9 and T10. CONCLUSION: Desflurane elicits a dose dependent increase in cerebral blood flow velocities and heart rate, but does not change mean arterial pressure, suggesting that its cerebrovascular action is independent of its systemic vascular action. CO2 reactivity is maintained at one MAC. The results in children are similar to those seen in adults. PMID- 9750735 TI - -Secondary cerebral stress of systemic origin in children with severe craniocerebral injuries-. AB - OBJECTIVE: To assess incidence of secondary brain insults of systemic origin (SBISOs) such as arterial hypotension, hypoxaemia, hypercarbia, and anaemia in severely head injured children; to assess their impact on mortality and morbidity in the short- and long-term. STUDY DESIGN: Prospective, open study covering a 24 month period. PATIENTS: Seventy-one children, under 15 years of age, admitted to a trauma centre for severe brain injury. METHOD: Analysis of SBISOs and outcome. RESULTS: Twenty-five children were admitted with SBISOs. The mortality rate was 37%. After hospitalization, 84% of the children with SBISOs vs 46% without SBISOs had severe disability (Glasgow outcome score = 1, 2 and 3). After 1 year, 20 out of the 45 children still alive were contacted. One of the four with SBISOs communicated a bad recovery. Fifteen children without SBISOs presented good recovery: GOS = 4-5, paediatric overall performance category (POPC scale) = 1-2. CONCLUSION: Hypotension was associated with significant increase in mortality (x 3.6) in children with severe head injury. The consequences were worse when anaemia was associated. PMID- 9750736 TI - -A difficult intubation of an infant with McKusick-Kaufman syndrome. Failure of the laryngeal mask-fibroscope sequence-. AB - Fibreoptic intubation through the laryngeal mask airway is a recommended technique for the management of a restricted airway. We report the failure of this technique in a 20-month-old infant presenting with McKusick-Kaufman syndrome. PMID- 9750737 TI - -Cerebral air embolism after removal of an internal jugular vein catheter-. AB - Central venous catheters are usually inserted and manipulated by anaesthetists intensivists and others familiar with their use under surgical conditions, yet they are often removed on the wards by junior doctors or nurses insufficiently trained in the removal procedure. In order to illustrate the risks presented by such a practice, we report a case of cerebral air embolism following the withdrawal of an internal jugular catheter in a sitting patient. The mechanisms of air entry into the venous and systemic circulation are considered, as well as the preventive and therapeutic measures. PMID- 9750738 TI - -Pancreatitis after blunt injuries to the abdomen-. AB - Three cases of pancreatitis occurring after a trauma to the pancrease are reported. They emphasize the difficulty of diagnosis at the initial phase of the condition. In all cases, computerized tomography (CT) scan was the main diagnostic method. Applying the same therapeutic strategy for pancreatitis as for other aetiologies facilitated a favourable outcome. PMID- 9750739 TI - -Severe self-poisoning with formol-. AB - We report a case of voluntary poisoning with formalin in a 47-year-old man. The initial status included respiratory failure, metabolic acidosis and coagulopathy. Medical therapy consisted of mechanical ventilation, a single prolonged haemodialysis session, N-acetylcysteine, and folic acid administration. The corrosive damage to the gastrointestinal tract required an oesogastrectomy and three months later a colic transplant. PMID- 9750740 TI - -Curative and preventive antibiotic therapy in infective endocarditis-. AB - Durack's criteria, including echocardiographic manifestations, are the current standard for the diagnosis of infective endocarditis (IE). The most common microorganisms known to cause IE are streptococci and staphylococci, and therapeutic principles are based on an association of parenteral antibiotics, as far as possible bactericidal and prolonged. Treatment also includes the search for the source of infection and its eradication. IE with negative blood cultures requires special techniques to obtain the causal microorganisms. In about half of the cases, a nosocomial bacteriaemia results in IE in patients with a prosthetic valve. Surgery is mandatory in IE with complications and/or caused by particular microorganisms; surgery is essential in most patients with a prosthetic valve. Although the presence of specific links between some procedures and the occurrence of IE has not been clearly proven, a prevention policy is nevertheless justified, considering the morbidity and mortality. Prophylaxis is indicated in patients with the cardiac conditions at risk for IE. IE prophylaxis prevails over prophylactic antibiotics usually administered for surgery. PMID- 9750741 TI - [The first death due to inhalation anesthesia in France. An example of the absence of verification of bibliographic references]. AB - The reporting of the first death attributed to anaesthesia is commonly placed in 1848. The present work indicates that, in France, anaesthesia was recognized as being responsible for deaths as early as February, 1847. PMID- 9750742 TI - -Coagulopathy suggestive of a primary fibrinolysis after head injuries with brain death-. AB - Coagulopathies associated with severe head trauma are usually of disseminated intravascular coagulation type with secondary fibrinolysis. We report a case whose semeiology was in part suggestive of a primary fibrinolysis. PMID- 9750743 TI - -The value of MRI in cerebral fat embolism-. AB - We report two clinical cases of cerebral fat embolism, thereby demonstrating the value of MRI. PMID- 9750744 TI - -Postoperative analgesia after major surgery of the knee. PMID- 9750745 TI - -An original device for the induction of inhalation anesthesia in children. PMID- 9750746 TI - [Quality of blood salvaged in orthopedic surgery by washing swabs]. AB - OBJECTIVE: To establish the feasibility and safety of recuperating blood absorbed by swabs used during orthopaedic surgery. STUDY DESIGN: Open, prospective study. PATIENTS: Included were children undergoing potentially haemorrhagic orthopaedic surgery for whom intraoperative blood salvage seemed possible. Excluded were those with contraindications for this procedure such as septic surgery and cancer surgery. METHOD: Intraoperative swabs used within the surgical field were collected by a surgical assistant, also in charge of weighing and washing them. The liquid was collected by the aspiration system of a recuperation-washing machine (RWM). The salvaged red blood cells were collected and retransfused at the end of surgery. Several samples of the washing liquid of the swabs and salvaged blood were taken during the procedure. The correlation between the quantity of blood shed and salvaged was calculated. The biological and clinical tolerance of the transfusion was assessed. RESULTS: Twelve patients undergoing surgery for scoliosis have been included. An average of 278 mL of blood were salvaged. In the washed cell concentrates the haematocrit was 54% and the free haemoglobin concentration was 3.84 g.L-1. All the bacteriological tests were negative over the first 24 hours. CONCLUSION: Provided that a strict operatory protocol is followed, this study demonstrates the possibility of recuperating blood from swabs used during major orthopaedic surgery. PMID- 9750747 TI - [Extramucosal pylorotomy via the umbilical route under general anesthesia and para-umbilical cord: evaluation of intra- and postoperative analgesia]. AB - OBJECTIVE: To assess both intra- and postoperative analgesia in infants undergoing extramucosal pylorotomy via a circumumbilical incision under general anaesthesia with neither opioid nor muscle relaxant, associated with a paraumbilical block. STUDY DESIGN: Open prospective study covering a 1-year period. PATIENTS: The study included 32 infants (age = 1.1 +/- 0.7 months, body weight = 4,427 +/- 876 g). METHOD: General anaesthesia was induced with either thiopentone or halothane and, after tracheal intubation, maintained with halothane in a N2O-O2 50 vol% mixture. Para-umbilical block was obtained using 0.5 mL.kg-1 of 1% lidocaine with epinephrine. Pain was assessed using time course of respiratory rate, heart rate and mean arterial pressure. A change of more than 20% in one of these variables was considered as criterion for poor analgesia. RESULTS: Intraoperative analgesia was adequate in all patients but one, 3 minutes after incision. Surgical conditions were considered as being good or satisfactory in 76% and 24% of cases, respectively. Postoperative analgesia, assessed 1 and 6 hours after completion of surgery, was convenient in 90% of infants, the end of the action of the local anaesthetic resulting in a poor analgesia at the 6th hour in four of them. CONCLUSION: Provided a short bevel needle is used and basic safety rules of local anaesthesia are followed, the para-umbilical block provides adequate intra and postoperative analgesia in infants undergoing extramucosal pylorotomy via circumumbilical route. PMID- 9750748 TI - [Use of sevoflurane for surgery of pheochromocytoma]. AB - Resection of phaeochromocytoma is associated with sudden changes in arterial pressure. Pharmacodynamic and pharmacokinetic properties of sevoflurane are suitable for maintenance of haemodynamic stability. We report two cases of phaeochromocytoma resection using sevoflurane. Intraoperative hypotensive or hypertensive events have been rapidly controlled, most often only by adjustment of the end-expiratory fraction of sevoflurane. PMID- 9750749 TI - [Kidney procurement from a heart-arrest donor]. AB - Brain death occurred in a 47-year-old head trauma patient following 5 days of intensive care. The procedure for multiple organ procurement was initiated. Irreversible cardiac arrest had occurred during the period in the surgical intensive therapy unit. External cardiac massage and mechanical ventilation were maintained until in situ cooling of the kidneys was achieved. The patient was taken to the operating room for nephrectomy. Both kidneys were transplanted with a favorable outcome. Procurement of kidneys from non-heart-beating donors could increase the number of available grafts for transplantation. PMID- 9750750 TI - [Hemorrhagic surgery in two Jehova's witness children refusing programmed autotransfusion: a place for erythropoietin]. AB - We report two cases of haemorrhagic surgery in a 6-year-old and 16-year-old girl, respectively, whose parents were Jehovah's witnesses and therefore opposed to preoperative blood donation, but accepting intraoperative blood salvage. Erythropoietin and intravenous iron were administered preoperatively to increase red cell mass. Intraoperative blood salvage, including normovolaemic haemodilution and intraoperative autologous transfusion, avoided homologous blood transfusion. PMID- 9750751 TI - [Hemophagocytic syndrome: an underestimated cause of the syndrome of non infectious fever?]. AB - Haemophagocytic syndrome (HM) is a severe disorder with characteristic clinical and biological features. The case of a patient with malignant lymphoma who experienced HM during Herpes virus I pulmonary infection is reported. Main clinical, biologial and pathological findings are described. The risk of underestimation of this disease in patients requiring intensive care as well as the absolute necessity of an early diagnosis are emphasised. PMID- 9750752 TI - [Left ventricular diastolic function: physiology, physiopathology, evaluation, therapy, consequences of anesthesia]. AB - With the exception of cardiac surgery, the acute disturbance of the left ventricular diastole occurs mainly in the elderly. Today it represents 30 to 40% of congestive cardiac failures, however with a lower mortality than for acute systolic disturbances. Generally indicated are relaxation anomalies, proto mesodiastolic mechanism and problems with compliance, an indicator of the pressure/volume diastolic relationship. Invasive techniques remain the standard method. Doppler echocardiography is becoming increasingly important for the assessment of diastolic function. In most cardiopathies, relaxation anomalies occur early, whereas compliance disturbances are mainly associated with advanced cardiac diseases. During anaesthesia, adverse events (auricular fibrillation, hypovolaemia) may worsen a fragile situation. Anaesthetic agents, in particular volatile agents, act on the ventricular diastole. Long-term therapy of diastolic anomalies includes agents amending left ventricular hypertrophy. Emergency therapy has not yet been systematised. PMID- 9750753 TI - [Admission modalities of brain dead patients at Ile de France hospitals which do not harvest organs. Cooperative Group for Transplantation of Ile de France (GCIF)]. AB - In order to identify possible causes for the shortage in organ procurement today in France, a regional survey including 74 hospitals in the Paris area which are likely to receive brain dead patients (BDP) and in which there were neither harvesting nor transplantation activities was conducted. Of the 66 hospitals (89%) answering this survey, half of them were district general hospitals. In the 2 years before the survey, they received on average three BDP. Such a figure represents at least 10% of BDP seen in this area. For one half of the centres, care of these patients was difficult or impossible due to the available facilities. Organisational concerns were among the major problems raised by the transfer of these patients to harvesting centres. A preestablished geographical network would be of help for simplifying the transfer of these patients. Interestingly, about 50% of centres already had such links with a transplantation centre. This study provides information concerning logistics and possible points which could be improved in order to increase the number of BDP liable to be transferred to transplantation centres. PMID- 9750754 TI - [Noninvasive postoperative ventilation using a buccal mask]. AB - In this preliminary study we assessed the feasibility and efficiency of non invasive ventilation (NIV) with a buccal mask (NIV-BM), not fitted with a mouthpiece, in surgical patients in acute respiratory failure whose lungs could not be ventilated with a face mask. In the eight patients enrolled in the study, NIV-BM increased tidal volume, decreased respiratory rate and improved PaO2. In two patients the trachea had to be intubated and one of them died of septic shock. NIV-BM can act as a valuable alternative in surgical patients in acute respiratory failure in whom a face mask cannot be used because of leaks or bad tolerance. PMID- 9750755 TI - [Concerning the recommendations of the French Society of Anesthesia and Intensive care on the use of a bacterial and viral filter on the anesthesia circuit]. PMID- 9750756 TI - [The Bonain anesthetic mixture: a forgotten ancestor of Emla cream?]. PMID- 9750757 TI - [Quality of care in anesthesia: search for predictive factors of postoperative pain]. PMID- 9750758 TI - [A solution for facilitating locoregional anesthesia using a nerve stimulator]. PMID- 9750759 TI - [Locoregional anesthesia for ophthalmic surgery: unique episcleral injection (sub tenon) in the internal canthus]. PMID- 9750760 TI - [For or against controlled ventilation using a laryngeal mask?]. PMID- 9750761 TI - [Systematic leukocyte depletion: a not negligible surcost for the safety of accrued transfusional material]. PMID- 9750762 TI - [Taking care of acute pain in pre-hospital medicine]. PMID- 9750763 TI - [Accessory anesthetic circuits: pro or con?]. PMID- 9750764 TI - [Infections and the practice of anesthesia and recovery]. PMID- 9750765 TI - [Prolonged effect of midazolam with delayed postanesthetic recovery from the possible interference of famotidine]. AB - We report the case of a 79-year-old woman, in chronic renal insufficiency who recovered from anaesthesia after a delay of 24 hours, after flumazenil (Anexate) administration. She was given famotidine (Peptidine) the day before surgery. Midazolam was administered for premedication (5 mg per os) and for the induction of anaesthesia (2 mg intravenous). Among the various causes of delayed recovery in this elderly patient, an interaction between midazolam and famotidine is discussed. PMID- 9750766 TI - [Areactive unilateral mydriasis after diagnostic celioscopy under general anesthesia]. AB - A unilateral areactive mydriasis occurred in a 49-year-old ASA I woman after a coelioscopic procedure while anaesthesia was maintained for an associated procedure. The various causes of brain stem damage were considered. The inability to perform a complete neurologic assessment resulted in the indication of a cerebral computerised tomography (CT) scan. The diagnosis of Adie's tonic pupil was made a posteriori, taking into consideration: 1) complete recovery without neurologic deficit; 2) unremarkable CT scan; 3) previous transient episodes of unilateral areactive mydriasis. PMID- 9750767 TI - [Lethal portal venous gas after cardiopulmonary arrest]. AB - We report the case of a 51-year-old patient admitted after a transient cardiorespiratory arrest. The abdominal CT scan revealed the presence of hepatic portal venous gas. At laparotomy, a diffuse mesenteric ischaemia was diagnosed. The patient died from multiple organ failure in the subsequent hours. Necrotic bowel is associated with hepatic portal venous gas in 50% of the cases and the current mortality rate is 85%. Gas originates either through intestinal transmucosal passage, either by intraportal bacterial gas production, or through both mechanisms. PMID- 9750768 TI - [Pros or cons of accessory anesthetic circuits. I. Arguments for their use]. AB - In addition to the circle breathing system, which represents the main circuit of the anaesthetic machine, the use of an accessory breathing system (ABS), either a partial rebreathing system according to Mapleson's classification, or a system including a non-rebreathing valve, is appropriate for the anaesthetic management of many patients, depending on their physical status, age, indication and duration of surgery. The same safety rules, namely full checking procedure before use of the system and monitoring of inhaled gases and end-tidal CO2 must be applied as for the main circle system. Potential complications resulting from non compliance with these rules cannot be considered valuable reasons for denying the use of breathing systems that have safely been used for decades in millions of patients. PMID- 9750769 TI - [Pros and cons of adding an accessory breathing system to the main circle circuit. II. Arguments against their use]. AB - When compared to the circle system alternative breathing systems (ABS) are of no benefit. When the only indication of an ABS is emergency oxygen administration it should be connected to the O2 pipeline upstream from the flowmeter bank and the vaporiser. The use of an ABS for anaesthesia maintenance is no longer justified because of the difficulties in monitoring pressure, flow and concentrations of the gas mixture, the cost of gas and vapour administered at a high flow and the resulting pollution. The use of an ABS for very short anaesthetics is only acceptable if the administered gas mixture is monitored. PMID- 9750770 TI - [Risk of nosocomial infection in anesthesia. General recommendations]. PMID- 9750771 TI - [Maintenance and disinfection of equipment necessary for airway control and mechanical ventilation]. PMID- 9750772 TI - [Evaluation of aseptic measures in the performance of epidural catheterization and perception of its risk of infection. Results of a survey in Languedoc Roussillon]. AB - OBJECTIVE: To assess the level of aseptic practice in epidural catheterization, to determine the perception of its infectious risk and the knowledge of epidural abscess symptoms. STUDY DESIGN: Cross-sectional regionwide retrospective survey. MATERIAL: Questionnaire sent after telephone call to 160 anaesthesiologists performing epidural anaesthesia in Languedoc-Roussillon. RESULTS: Replies were obtained from 96 anaesthesiologists working in 26 centres. Handwashing techniques were various: 53% onefolded with an antiseptic preparation, 38% twofolded and 9% used a simple soap; 41% did never wear a long-sleeved sterile surgical gown and only 53% wored simultaneously a cap; a surgical mask and a sterile gown; 4% of the Tuohy needles were reusable; 21% of the dressings permitted continuous inspection of the site of puncture; 12.5% admitted not to know epidural abcess symptoms and 30% ignored the most adapted radiological diagnostic means. CONCLUSIONS: Good aseptic practice in epidural catheterization should be clarified by a consensus conference. There is an obvious lack of knowledge concerning features of epidural abscess. PMID- 9750773 TI - [Ethical aspects concerning nosocomial infections]. AB - Ethical issues about nosocomial infections have to be considered in two fields: the daily practice and the doctors participation in the institutional or suprainstitutional committees, involved in various administrative, strategic or financial measures aiming at the control and the prevention of nosocomial infections. In daily practice the ethical rules are founded on the principles of individual good: regarding nosocomial infection the principle of non maleficence is the most relevant. Physicians, nurses or other health professional may have a part of responsibility in a nosocomial infection. However there are many impediments to their acknowledgement of their own moral responsibility. The most important impediments may be: a) the more and more collective approach of care in many hospitals wards; b) the fact that the consequences of a nosocomial infection in one patient can extend to patients in other structures and, thus, can remain ignored by those who are responsible for this infection; c) paradoxically enough, the high attention paid to the theoretical issues concerning nosocomial infection. It must be kept in mind, too, that the ethical issues concerning nosocomial infections include the necessity for providing the patients with adequate and truthful information about the risks of nosocomial infection, in every hospital or ward, and--if it happens--about the nosocomial nature of an infectious complication. There is also some concern about the fact that the various modalities of legal responsibility and indemnification for a nosocomial infection, that the law has already specified or could define in a near future, may have a negative influence on the capacity of many care providers to keep a feeling of their moral responsibility. In the official committees devoted to decide administrative, strategic or financial measures able to help scientific research and to induce better practice in the surveillance and the prevention of nosocomial infections, the ethical principles at use are not only those of good but also those of justice. The balance between these ethical principles is delicate, however, for doctors, the main concern should be to avoid any distortion of the debate which could result from their confusing use of the principles of good. PMID- 9750774 TI - [Evaluation of professional development of Cambodian male nurses specializing in anesthesia and intensive care, trained through humanitarian care programs]. AB - OBJECTIVES: To assess the working conditions of Cambodian male nurses specialised in anaesthesia and intensive care (NSAIs), degree of satisfaction, whether training was suited to the Cambodian needs and practical application of training. STUDY DESIGN: Prospective survey. PERSONS: Two training years including 30 NSAIs. METHOD: External assessors evaluated working conditions, practice of anaesthesia, analysed logbooks and theatre reports, organised semi-directive interviews and examinations using clinical cases. RESULTS: Out of the 30 NSAIs, 28 had an appointment, mainly in anaesthesia (80% of their activity) and three-quarters of them felt that their skills were appreciated by their superiors. Seventeen had some form of responsibility in the management of a department. For the administration of an anaesthetic, 13 NSAIs of the second year had achieved an acceptable level of performance and resolved effectively 85% of the submitted cases. Twenty-two NSAIs reported difficulty in applying techniques learned during their training to real working conditions. The causes were poor equipment, poor organisation and poor relations with the hierarchy. The latter cause decreased the capacity to take appropriate decisions, which was the most common error made by the second year NSAIs. Finally, as their wages remained unchanged, 19 out of the 28 NSAIs were obliged to look for an additional source of income. CONCLUSION: This survey shows the medium-term effectiveness of an NSAI training programme basing on teaching and public health principles and organised in the humanitarian aid sector. PMID- 9750775 TI - [Locoregional anesthesia of the lower limb at admission of a polytrauma patient with a leg fracture]. PMID- 9750776 TI - [Streptococcus group A toxic shock. Beneficial effect of high flow hemofiltration, of short duration]. PMID- 9750777 TI - [Jehovah's witness: a case of a traffic accident with a fatal outcome]. PMID- 9750778 TI - [Locoregional anesthesia in ophthalmology: the matter of indications and techniques]. PMID- 9750779 TI - [Paravertebral space block in children]. PMID- 9750780 TI - [Problems posed for the anesthetist by Duchenne muscular dystrophy]. PMID- 9750781 TI - [How should one create an ambulatory surgery facility?]. PMID- 9750782 TI - [The "CE" stamp on ISO norms]. PMID- 9750783 TI - [Copa (cuffed oropharyngeal airway)]. PMID- 9750784 TI - [Frequent preoccupation with the use N2O]. PMID- 9750786 TI - [Management of postoperative pain in adults and children]. PMID- 9750787 TI - [How is postoperative pain evaluated?]. AB - The assessment of postoperative pain and analgesic efficacy is essential as pain levels and morphine requirements are not predictable. Self-assessment with unidimensional methods (such as the visual analogue pain scale, the numerical rating scale and the verbal rating scale) is the rule for adults and children more than 5 years of age. The former is a validated method and the most accurate and reproducible scale. Assessment of pain is difficult in children less than 5 years old. Only the scales for hetero-assessment with behavioural assessment (CHEOPS and OPS) are used. Finally, morphine consumption with PCA is also an indirect pain assessment method. Postoperative pain should be assessed several times a day in every patient, starting in the recovery room and prolonged during hospital stay. Pain should be measured at rest and in dynamic conditions by the medical and paramedical team. PMID- 9750788 TI - [Frequency, intensity, development and repercussions of postoperative pain as a function of the type of surgery]. AB - Type of surgery is the most important factor conditioning intensity and duration of postoperative pain. Thoracic and spinal surgery are the most painful procedures. Abdominal, urologic and orthopedic surgery lead to severe postoperative pain. Duration of severe pain rarely exceeds 72 hours. Mobilization increases pain intensity after abdominal, thoracic and orthopaedic surgery. Pain could occur after daycase minor surgical procedures and is often underestimated. Postoperative complications related to pain are difficult to disclose because of the interposition of the direct effects of analgesic treatments. Respiratory and cardiovascular postoperative complications are unrelated to postoperative pain in healthy subjects. This could be different in high risk patients. The surgical procedure is the major determinant of metabolic and psychologic postoperative deterioration. Adequate pain relief allows postoperative rehabilitation and physiotherapy programmes after abdominal and orthopaedic surgery. This could be expected to reduce hospital stay and improve convalescence. PMID- 9750789 TI - [How is the management of postoperative pain in organized in surgical wards?]. AB - Many studies have demonstrated that the management of pain after surgery was unsatisfactory. New pain management techniques have been developed in recent years (patient-controlled analgesia, epidural analgesia). To extend the number of patients who may benefit from these recent techniques and/or to obtain the best efficacy from existing methods of pain relief, re-organisation should take place on surgical wards. For example, protocols describing pain management strategies should be written. Surveys and audits should be carried out regularly to check their efficacy. Moreover, patients should be fully informed of the range of treatments available and their adverse effects. Finally, all staff involved in providing acute pain relief should undergo training. PMID- 9750790 TI - [Benefit-risk and monitoring modalities of different techniques and methods of postoperative analgesia]. AB - This review aimed to determine the benefits-risks ratio of postoperative analgesia. The various agents usually used for intravenous postoperative analgesia (paracetamol, NSAID's, opioids), and the techniques for postoperative analgesia (PCA, epidural, perinervous block) are analysed. The rules proposed for the monitoring of postoperative analgesia are considered. PMID- 9750791 TI - [Does a means exist for prevention of postoperative pain?]. AB - Postoperative pain can be prevented through pharmacological and non pharmacological means. The influence of the interval between therapy and surgical stimulus, which corresponds to "pre-emptive analgesia" in English-speaking countries (comparison of preoperative administration with pre- and postoperative administration of the same analgesic) has been assessed by numerous studies of good methodological quality. In spite of the initial promising results, most of the results published at present are negative. In an enlarged concept of postoperative pain prevention, various trials have demonstrated the benefits of the choice of the surgical approach, as well as the psychological preparation of the patient for surgery. PMID- 9750792 TI - [The epideiology of postoperative pain]. AB - Postoperative pain, as all types of pain, is a complex phenomenon including sensory, emotional and behavioural factors. The incidence and severity of postoperative pain is very variable between patients and is rather unpredictable. Patients characteristics as well as the types of surgery and anaesthesia will be of importance. Health professionals have a major role to play for improving effectiveness of pain management as well as safety. PMID- 9750793 TI - [Repercussion of postoperative pain, benefits attending to treatment]. AB - Physiological responses to postoperative acute pain may impede organ functions (cardiovascular, pulmonary, coagulation, endocrine, gastrointestinal, central nervous system, etc). Pain alleviation improves patient's comfort, but also may minimise perioperative stress response, physiological responses and postoperative organ dysfunction, assist postoperative nursing and physiotherapy, enhance clinical outcome, and potentially shorten the hospital stay. Potent postoperative analgesia, especially by epidural route, may be associated with reduction in incidence and severity of many perioperative dysfunctions. Peridural analgesia using local anaesthetics is the best technique for decreasing postoperative stress after lower abdominal or lower limb surgery. Analgesia using either epidural or high doses of morphine may improve some cardiac variables such as tachycardia and ischaemia, but does not change the incidence of severe cardiac complications. For patients undergoing vascular or orthopaedic surgery, epidural analgesia can improve clinical outcome by preventing the development of arterial or venous thromboembolic complications. However, in comparative studies, the control groups did not receive adequate prophylactic treatment for thromboembolic complications. Epidural analgesia can hasten the return of gastrointestinal motility and shorten the hospital stay. Postoperative mental dysfunction is decreased using intravenous PCA morphine in the elderly. Epidural analgesia with local anaesthetics improves postoperative respiratory function but, for unknown reasons, these benefits are not associated with a decrease in respiratory complications. On balance, the mode of acute pain relief decreases adverse physiological responses and many intermediate outcome variables; however, there is inconclusive evidence that it affects clinical outcome. Major advances in postoperative recovery can be achieved by early aggressive perioperative care, including potent analgesia, early mobilisation and oral nutrition. As a result, the hospital stay may be shortened. PMID- 9750794 TI - [Evaluation of postoperative pain]. AB - Pain is a subjective feeling; its assessment is therefore difficult, and no "gold standard" method exists for humans. Major improvements have, however, been made in the last decade by widespread acceptation of the concept of pain evaluation and widespread use on surgical wards. Evaluation by the patient himself is the rule (unless communication is impaired), as assessment of pain by nurses or doctors systematically leads to underestimation (which also occurs with observational scales). Theoretically, pain should be evaluated in its multiple dimensions such as intensity, location, emotional consequences and semiologic correlates. Scales which have been developed to evaluate these dimensions are, however, too complex for widespread and repetitive use in surgical patients. The Mac Gill Pain Questionnaire is therefore only used in the surgical setting for research purposes. Moreover, its scientific accuracy, although often accepted, is poor and in our opinion cannot be accepted as a reference method. Only methods assessing pain intensity can be used in the clinical setting because of their simplicity. The verbal rating scale (VRS), the numerical rating scale (NRS) and the visual analogue scale (VAS) are preferred by an increasing number of groups. Although scientific validation is difficult, VAS seems the most accurate and reproducible scale. Post-operative pain should be assessed several times a day in every patient, at rest and in dynamic conditions (cough, movement) and should focus on present pain rather than on pain in the previous hours. Assessment of pain is essential before quality-assurance programmes can be implemented. PMID- 9750795 TI - [Conventional techniques for analgesia: opioids and non-opioids. Indications, adverse effects and monitoring]. AB - Morphine dosage must be carefully adapted in patients with renal failure or severe liver failure. The i.v. route is used for morphine titration in the post anaesthesia care unit (PACU), or for analgesia in children. Systematic (not on demand) intramuscular or subcutaneous morphine must be administered at intervals not longer than 4 hours. Dosage is best determined after i.v. titration in the PACU. Codeine, administered orally, is metabolised into morphine. Codeine has almost no effect in 7% of Caucasians and at least 15% of Asians. Nalbuphine, which has a sedative effect and a short half-life, is mainly used in children. Paracetamol (acetaminophen) is used orally or rectally, most often in combination with codeine. Paracetamol dosage is 60-90 mg.kg-1.d-1, including a 20 mg (orally), or 40 mg (rectally) loading dose. Its therapeutic ratio is low, with a potential hepatic toxicity. Dosage must be lowered in alcoholics or in patients under isoniazide therapy. Non-steroidal anti-inflammatory drugs are powerful antinociceptive agents. Their use must be restricted to the first 5 postoperative days. Their major contraindications are kidney failure, risk of gastrointestinal bleeding, coagulation disorders, allergy. They also have a marked morphine sparing effect and reduce therefore the respiratory depression induced by morphine. PMID- 9750796 TI - [Patient-controlled analgesia. Benefits, risks, methods of monitoring]. AB - Patient-controlled analgesia refers to a relatively new approach to morphine delivery in which patients are allowed to self-administer small doses of an opioid, to achieve adequate relief of postoperative pain. The main benefit is to reduce fluctuations in opioid plasma concentrations. A matter for worry remains the occurrence of side effects, especially ventilatory depression. In order to guarantee the efficacy and safety of this technique, the education of patients and nurses is essential. Protocols are required, specifying the use of this technique (prescription, patient monitoring, treatment of side effects). PMID- 9750797 TI - [Postoperative locoregional analgesia in the adult: epidural and peripheral techniques. Indications, adverse effects and monitoring]. AB - Regional analgesia is a very effective way to treat postoperative pain. Lumbar and thoracic epidural analgesia are well adapted to major abdominal and thoracic surgery. Nevertheless, respiratory side effects induced by opioids are potentially severe and an adequate monitoring is essential. In orthopaedic surgery, perineural blocks are the best technique to manage postoperative pain and perineural catheters may be used. The importance of intra-articular analgesia, simple and safe, is not fully understood. The association of a local anaesthetic inducing a minor motor block and a strong sensitive block (bupivacaine, ropivacaine), with an opioid seems to be the best pharmacologic choice regarding quality of analgesia and safety. Benefits of postoperative regional analgesia on mortality and morbidity are not demonstrated. Medical and nursing staff and specialized units should improve quality of postoperative regional analgesia as well. General guidelines for the practice of regional anaesthesia must be closely followed. PMID- 9750798 TI - [How can pain of surgery be limited?]. AB - Postoperative pain is due to direct stimulation of nociceptors by surgical trauma, and by algogenic substances produced by damaged tissues. Control of surgical pain can be obtained by limiting the extent of damage to tissues as well as the choice of incision. Endoscopic or video-assisted surgery is an effective mean to reduce pain caused by surgical approach. It is widely used in abdominal, thoracic, orthopaedic surgery, and urology. Many studies have shown a reduction of postoperative pain by laparoscopy for gynaecological surgery and cholecystectomy, but for other procedures the potential advantage of laparoscopic surgery has not yet been established. Conventional open surgery is still widely used. It has been suggested that transverse laparotomies are less painful than midline incisions, and that incision by electrocautery was less painful than with scalpel; but this has not been strictly established. Infiltration of wounds or nerves with local anaesthetic agents is a way of clinical research, which merits further investigation. Whether delicacy in surgery is capable of minimising pain by limiting tissue attrition remains to be demonstrated. Finally, drains and catheters, particularly the naso-gastric tube, which are responsible for pain, could be abandoned when not essential. PMID- 9750799 TI - [Prevention of postoperative pain]. AB - The pre-emptive analgesia concept suggests that pre-administration of analgesics may enhance the efficacy of these drugs. This review has selected the data from the literature according to two types of methodological criteria: Sackett's criteria, and those specific of pre-emptive analgesia studies. Infiltration, spinal and peripheral nerve blocks using local anaesthetic drugs do not seem to produce pre-emptive analgesia. The few positive results have limited clinical significance. The results concerning opioids are contradictory and the clinical significance is limited. Preoperative oral administration of non steroidal anti inflammatory drugs (NSAIDs) offers no benefit. Intravenous pre-administration has a limited advantage, but enhances perioperative bleeding. Ketamine, an NMDA receptor antagonist, may have some pre-emptive analgesic properties according to the few studies available. In conclusion, pre-administration of analgesic drugs represents the usual strategy for the anaesthesiologist (spinal or peripheral block, infiltration, opioids). In other cases (NSAIDs, ketamine), pre administration represents a change in usual practice. This is not justified for NSAIDs; NMDA receptor antagonists may offer an interesting research area. Data concerning pre-emptive analgesia for chronic pain syndrome such as phantom limb pain are quite limited. PMID- 9750800 TI - [Postoperative pain. Particularities in the child of less than 5 years, neonatology excluded]. AB - For many years, postoperative pain has been undertreated in children less than 5 years old in comparison to adults. The assessment of pain is indeed difficult in this range of age, and only the scales of hetero-evaluation are used. The guidelines for treatment are similar as in adults: systematic administration, balanced analgesia, evaluation of pain and potential adverse effects. Non opioid analgesics used are mainly paracetamol, niflumic acid and ibuprofen. Morphine remains the drug of choice among opioids; however the risk of respiratory depression in higher in infants less than 3 months old. Nalbuphine is also widely used in paediatrics. In addition, regional anaesthesia, either in single shot for minor surgery, or in continuous administration through epidural catheter for major surgery, has changed the management of postoperative pain in paediatrics. PMID- 9750801 TI - [Postoperative analgesia. Specificity in the elderly]. AB - The necessity of an adapted, optimal postoperative analgesia in the elderly is widely recognised. Reduced physiological capacities must be taken into consideration during the perioperative period. Class I analgesics, such as paracetamol, are both safe and efficient, and can be used for basic analgesia. Non steroid anti-inflammatory drugs carry an increased iatrogenic risk in the elderly. Their benefits should always be considered with regard to their risk. Their dosage should be decreased by 40-60% in comparison to the standard adult doses. Opioids, though highly efficient, carry a higher risk of respiratory depression due to the increased sensitivity to this class of molecules in the elderly. Doses must be reduced by 50% of the standard adult dose in order to limit adverse events while maintaining an equivalent level of analgesia. Patient controlled and spinal opioid analgesia can be used in elderly patients. However surveillance of both the state of consciousness and respiratory rate must be carried out hourly over a period ranging from 12 to 24 hours. Pulse oximetry can be of value. After orthopaedic surgery, perineural or peripheral analgesia should be favoured considering the excellent benefit-risk ratio. Close clinical monitoring is essential for providing safe and efficient analgesia in the elderly using the techniques currently at our disposal. PMID- 9750802 TI - [Management of postoperative pain in surgical units]. AB - In order to improve the management of postoperative pain many publications insist on progressive changes in care organization. The following list outlines steps to be taken for implementation of these changes: 1) an initial analysis of management of post-operative pain allows awareness of reforms to be proposed; 2) participation of health teams in special training in order to use evaluation tools and collect data (use of analgesics, adverse effects); 3) establishing policies and procedures: recovery room, guidelines for analgesic use and adverse effects; 4) notifying patient about the various procedures to be used in postoperative period--discussion with the patient during the preoperative interview; 5) current use of standard patient-controlled analgesia (PCA) and locoregional analgesia; 6) use of combined techniques in order to achieve a balanced analgesia; 7) implementing a quality assurance programme which should include analgesic effectiveness, patient satisfaction and prevention of complications; and 8) planning of an Acute Pain Service based on a clinical nurse co-ordinator which offers highly effective forms of postsurgical analgesia. PMID- 9750804 TI - [Evaluation of training on intubation with a rigid fiber optic laryngoscope (UpsherScope)]. AB - OBJECTIVE: To evaluate the learning of tracheal intubation with a new rigid fibreoptic laryngoscope (UpsherScope). STUDY DESIGN: Open prospective study. PATIENTS: Five investigators used the UpsherScope to intubate the trachea in 164 patients scheduled for gynaecological surgery requiring tracheal intubation. All patients were of physical class ASA I or II and criteria for difficult intubation were negative. METHODS: After muscle relaxation, 120 seconds were allowed to intubate the trachea with the UpsherScope. If intubation had not been achieved by that time, the attempt was considered as a failure and the trachea was intubated using conventional laryngoscopy. RESULTS: The overall success rate with the UpsherScope was 73%. Forty-five tracheas could not be intubated with the device within 120 seconds. The inability to insert the tracheal tube through the vocal cords despite a good view of the larynx (23/45) or the inability to visualise the glottis because of secretions (21/45), were the two main causes of failure. CONCLUSION: The UpsherScope, a new rigid fibreoptic laryngoscope devised for routine and difficult intubation, is robust and allows the view of the tracheal tube passing between the vocal cords. However, in this study the intubation success rate remained low and was not improved by further experience. No benefit was found with the UpsherScope in patients with normal airways. Further studies are necessary to assess its efficiency in cases of difficult intubation. PMID- 9750805 TI - [Brachial plexus anesthesia via an axillary route for emergency surgery: comparison of three approach methods]. AB - OBJECTIVES: To compare three techniques of brachial plexus blockade for emergency surgery of the upper limb. STUDY DESIGN: Prospective, randomised study. PATIENTS: One hundred eleven patients admitted to an emergency surgical service, randomly assigned to three groups. METHODS: The patients were given 2% lidocaine with epinephrine 20 mL and 0.5% bupivacaine 20 mL. The three groups were as follows: brachial plexus block using a peripheral nerve stimulator (group St, n = 38); transarterial brachial plexus blockade with injection of 2/3 of the anaesthetic in back of and 1/3 in front of the artery (group TAP, n = 36); transarterial brachial plexus blockade with one single injection in back of the artery (group TP, n = 37). The success rate, time required to perform the technique, latency of analgesia, quality of motor blockade, and adverse effects were compared between the three groups. Analysis of variance was used to compare quantitative data and chi 2 test were used for qualitative data. RESULTS: Rates of success varied between 65 and 75%. Success rates, latency of analgesia and quality of motor blockade were not significantly different between groups. Time to perform the technique was longer when using a nerve stimulator. CONCLUSION: As these three techniques for brachial plexus block in emergency surgery are comparable, no one can be recommended instead of the others. PMID- 9750806 TI - [Snake bite by European vipers. A multicenter study of tolerance to Viperfav, a new intravenous antivenom]. AB - OBJECTIVES: To evaluate the tolerance and the effectiveness of i.v. Viperfav, a new antivenom containing F(ab')2 fragments of equine antibodies, for the treatment of European viper envenomed patients. STUDY DESIGN: Open, multicentre field trial, associated with a cohort study. PATIENTS: The study included 46 patients of either gender, nine aged less then 10 years, eight between 10 and 15 years, and 28 adults, who sustained a moderate or severe viper envenomation (Grade 2 or 3). METHOD: At the inclusion, a single infusion of Viperfav was given. Depending on the clinical course, up to four additional infusions were to have been administered at 4-hour intervals. To evaluate tolerance, all symptoms were recorded. There were three effectiveness evaluation criterion (duration of hospitalisation, course of the severity grade, recovery (sequelae)) and one subjective criteria (value of the antivenom as ascertained by investigators). RESULTS: In the 46 included patients, 79 infusions were administrated. Concerning tolerance, six mild symptoms were associated to the antivenom infusions. No severe reaction occurred. The mean duration of hospitalisation was 4 days 19 hours +/- 13 hours. A severity grade decrease by at least one point was observed in 35 patients, and all were discharged without sequelae. For the investigators the antivenom was inefficient in only two patients (grade 3 with tissue lesions). CONCLUSIONS: In comparison with literature data (5 to 10% of severe reactions attributable to the antivenom), the tolerance of Viperfav can be considered as satisfactory. As all criteria were in favour of a positive benefit to risk ratio, the authors recommend the use of Viperfav i.v. for the grade 2 and 3 envenomations instead of the current less purified antivenom, which can only be administered by the intramuscular route. PMID- 9750807 TI - [Rapid sequence anesthetic induction via prehospital tracheal intubation]. AB - The choice of sedation for emergency intubation remains controversial. This lack of consensus has led to various sedation protocols used in French prehospital care setting. A review of data from the literature suggests that the association etomidate-suxamethonium is probable the best choice for rapid sequence intubations in the prehospital setting. Its benefits include protection against myocardial and cerebral ischaemia, decreased risk of pulmonary aspiration, and a stable haemodynamic profile. Randomized studies are needed to substantiate the advantages of the association etomidate-suxamethonium for rapid sequences intubation in the prehospital setting. PMID- 9750808 TI - [Heat and moisture exchanging filters for conditioning of inspired gases in adult anesthesia and resuscitation]. AB - OBJECTIVES: Heat and moisture-exchanging filters (HMEFs) are increasingly used in clinical practice. At the same time, new scientific data are available which clarify the benefits of these devices. DATA SOURCES: We searched in the Medline database for all papers written in English or French, without limiting date of publication, using the following key-words separately or in combination: humidity, temperature, mechanical ventilation, equipment. STUDY SELECTION: From the 200 articles provided by Medline, we selected those directly concerning HMEFs. Some older studies and those on HMEFs no longer available were excluded. DATA EXTRACTION: Principle data available from the literature were analysed. DATA SYNTHESIS: Humidification and warming of the inspired gas mixture is mandatory during mechanical ventilation. There is a direct link between HMEF performance and the characteristics of tracheal secretions. This justified the recommendation for the use of HMEFs with a humidity output above 30 mg of water per litre of gas mixture. In this case, HMEFs are as efficient as conventional heated humidifiers. HMEFs seem to decrease the rate of nosocomial pneumonia in comparison with heated humidifiers. HMEFs induce a slight increase of dead space which should be taken into consideration during weaning from mechanical ventilation. There are demonstrable data in the literature suggesting the possibility of cross viral infection via the anaesthetic machine when an HMEF is not used. There are no data which suggest a specific type of HMEF regarding viral filtration. CONCLUSION: According to the literature data, using an HMEF is essential in anaesthesia and is highly recommended in intensive care. PMID- 9750809 TI - [Claude Bernard-Horner syndrome and its opposite, Pourfour du Petit syndrome, in anesthesia and intensive care]. AB - OBJECTIVE: To analyse cases of Horner's syndrome (HS) and its opposite, Pourfour du Petit's syndrome (PPS), occurring in anaesthesia and intensive therapy with consideration of the data of current literature. DATA SOURCES: For this paper we have reviewed the French, English and German literature published in anaesthesia and intensive care journals using Medline search and the current textbooks. STUDY SELECTION: All observational studies on these syndromes, whether clinical cases or letters to the editor, form the basis for this article. DATA EXTRACTION: The articles were analysed mainly with regard to diagnosis, therapy and prognosis of syndromes due to iatrogenic causes. DATA SYNTHESIS: HS is caused by a paralysis of the ipsilateral sympathetic cervical chain and includes a ptosis of the upper eyelid, a slight elevation of the lower lid, a sinking of the eyeball, a constriction of the pupil, a narowing of the palpebral fissure, a nasal stuffiness associated with anhidrosis, and flushing of the affected side of the face. Regional anaesthesia (intra-oral anaesthesia, brachial plexus block, epidural anaesthesia whether by thoracic, lumbar or caudal approach, as well as interpleural analgesia) is the main anaesthetic cause for HS. HS due to the effect of a local anaesthetic is transient, it can precede a high spinal block and a cardiovascular collapse. HS from puncture of the internal jugular vein is most often permanent. When transient, HS regresses within 3 months after puncture. Other causes of HS include intraoperative posture, pleural drain, neck surgery, neck trauma. A mydriatic collyrium, such as phenylephrine, resolves ptosis for less than 1 hour and results in blurred vision from pupillary dilation. Major ptosis requires surgery. PPS is the reciprocal HS and is caused by a stimulation of the ipsilateral sympathetic cervical chain. PPS can precede HS. It carries a risk for conjunctivitis, keratitis and epiphora in case of major exophthalmia. PPS is often reported as an unilateral mydriasis. PPS has the same causes as HS. Myotic collyriums are relatively inefficient. Major lid retraction requires a tarsorraphy, pomades and nocturnal lid occlusion. A part of HS and most PPS occurring in anaesthesia and intensive care remain unrecognized or are recognized with delay, especially if they remain minor and transient or when they occur in unconscious patients, in horizontal posture. PMID- 9750810 TI - [Prolonged neuromuscular blockade with mivacurium in a newborn]. AB - Mivacurium is a short acting non-depolarising neuromuscular blocking agent. Short duration of action is due to a rapid hydrolysis by plasma cholinesterase (CHe). The duration of neuromuscular blockade can be prolonged by an abnormal variant of CHe. We report a case of a newborn with neuromuscular blockade for a duration of 8 hours following mivacurium 0.2 mg.kg-1. CHe activity values were not contributive for the diagnosis. The diagnosis was obtained with molecular study showing the new-born homozygocity. The whole family was heterozygous. This case emphasises the lack of precision of CHe activity measurement during the first 6 months of life. PMID- 9750811 TI - [Massive gas embolism following lung inflation for thoracic tomodensitometry in a multiple trauma patient with lung contusions]. AB - We report a case of gas embolism into both right and left circulation in a polytrauma patient with lung contusions, revealed by thoracic CT scan showing the heart and aorta filled with gas. It followed a lung inflation with a O2/N2O mixture for about 30 seconds at a pressure of at least 40 cmH2O in order to obtain apnoea for CT scan and to recruit atelectatic territories. The presumed mechanism was the passage of the O2/N2O mixture during the lung inflation manoeuvre out of disrupted airways into torn pulmonary blood vessels and pushed back into the heart chambers. The patient recovered fully. Lung inflation manoeuvre to obtain a prolonged apnoea during CT scan examinations of thorax is contraindicated in case of thorax trauma, as it carries a risk of gas embolism. PMID- 9750812 TI - [Cardiac arrest in an adolescent with arrhythmogenic right ventricular dysplasia]. AB - We report the case of a 16-year-old girl who experienced sudden cardiac arrest from ventricular fibrillation, complicating an arrhythmogenic right ventricular dysplasia, a rare heart muscle disorder, occurring typically in young adults, characterized by a fibrofatty replacement of the right ventricular myocardium. Symptomatic ventricular arrhythmias are frequent, and sudden death has been reported. In our case, diagnosis of arrhythmogenic dysplasia was based on the association of one major criterion and two minor criteria as suggested by the relevant task force. In contrast with most other reports, the chest ECG did not display the typical features. An automatic transvenous pectoral cardioverter defibrillator was implanted. The authors emphasise that juvenile forms are more exposed to ventricular fibrillation and sudden cardiac death, and consequently require the early detection of the disease. Family cases have been described and the occurrence in one individual must lead to investigations in the relatives. PMID- 9750813 TI - [Total spinal anesthesia after posterior lumbar plexus block]. AB - We report a case of total spinal anaesthesia which occurred after a lumbar plexus block using a posterior approach. After total hip arthroplasty under general anaesthesia, a lumbar plexus block was performed according to Winnie's landmarks at the L4 interspace using a nerve stimulator. Aspiration test for blood and spinal fluid were both negative, as well as a test dose of 3 mL lidocaine 2% bupivacaine 0.5%. One minute after the injection of 27 mL of the same mixture, a complete anaesthetic block occurred with hypotension and loss of consciousness requiring intubation and controlled ventilation during 3h30, without sequelae. Lumbar plexus block using a posterior approach must be performed cautiously and a slow and fractionated injection of the full dose is recommended. PMID- 9750814 TI - [Asphyxiating intra-alveolar hemorrhage: a rare form of fat embolism syndrome]. AB - We report a case of a 23-year-old patient admitted for a right femur fracture resulting from a traffic accident. An intra-alveolar haemorrhage occurred 48 hours later, with asphyxia anaemia, haematic bronchial aspirations, and bilateral alveolar opacities at chest X-ray. This symptomatology was associated with fever, sub-conjunctival petechiae, major hypocholesterolemia, deterioration of renal function, and cholestasis. All these features suggested a fat embolism. Other possible aetiologies were discarded because of normal cardiovascular and immunologic systems and absence of infection. The outcome under symptomatic treatment was satisfactory within 15 days. The occurrence of intra-alveolar haemorrhage in post-traumatic fat embolism is a rare event caused by pulmonary capillary obstruction by fat emboli. PMID- 9750815 TI - [Halogenated anesthetic vaporizers: importance of maintenance regulations]. AB - We report a serious dysfunction of 19 halothane vaporisers Vapor 19.3 (Drager) which delivered a much higher concentration of agent than indicated on the dial. This inaccuracy was linked to a major corrosion of the inner layers of the vaporiser, with a deposit of zinc bromide and chloride in the bypass channel. The main cause for this dysfunction was the absence of an adequate maintenance of the vaporisers since their purchase 3 years before. PMID- 9750816 TI - [Early perioperative mortality in a multidisciplinary hospital]. AB - OBJECTIVE: To evaluate the incidence and the causes of early intra- and postoperative deaths in a multidisciplinary hospital. STUDY DESIGN: Retrospective survey. PATIENTS: All patients receiving an anaesthetic between 1992 and 1995. METHODS: Analysis of all deaths occurring during anaesthesia and in the subsequent 24 hours. Demographic data (age, gender) and medical data (ASA physical class, type of surgery and degree of emergency) were recorded. The contribution of anaesthesia, surgery or patient disease to fatal outcome was analysed. RESULTS: The analysis included 52,654 patients who underwent either general anaesthesia or epidural analgesia. Perioperative mortality (n = 170) was 1/310 patients (0.32%). The risk factors for mortality (multivariate analysis) were: age > 64 years (odds-ratio [OR] 4.8), ASA class > or = 3 (OR 16.6), emergency surgery (OR 3.6), duration of surgery > 115 minutes (OR 37.4). Fifty percent of deaths (95% confidence interval [CI] = 42-58) were related to patient's underlying diseases and 29% to surgery (CI 95% = 22-36). The percentage of deaths linked to anaesthesia was 17.6% (CI 95% = 11.9-23.3, 1/1,755), consisting of 8.2% (CI 95% = 4.1-9.3, 1/3,761) totally due to anaesthesia and 9.4% (CI 95% = 5-13.8, 1/3,291) only partially. The main aetiologies of the deaths linked to anaesthesia were a mismanagement of severe haemorrhages (30%), respiratory complications (23%) or cardiac complications (23%). The mismanagement of an intraoperative critical situation (46%) and a mistake in the post-operative care (33%) were the main causes. DISCUSSION: In this survey, mortality due to anaesthesia was higher than the rates reported in other studies. Human error remained the main cause. PMID- 9750817 TI - [Survey of 39 South East Group hospital centers on the practice of antibiotic prophylaxis in surgery]. AB - A survey was carried out in 39 hospitals regarding the prescribing of perioperative antibiotics. The day of the survey, hospital pharmacists collected information on prescription patterns of antibiotics and, for 3 days, the duration of their administration. A total of 1,131 cases were surveyed, originating from digestive surgery (20%), orthopaedics (31%), gynaecology (15%), ophthalmic surgery (15%), and others (19%). Duration of surgery was 72 +/- 68 min. For all types of surgeries, antibiotics not recommended by the French consensus conference were prescribed. Third generation cephalosporins were used in 17% of patients in gynaecological surgery. In 20% of cases, antibiotics were administered at an inappropriate time: 9.5% after the beginning of surgery, 8% at the time of pre-medication, and 2.5% the day before surgery. Administration exceeded 48 hours in 10% of the cases. In conclusion, special attention should be paid to limit the prescription of third generation cephalosporins, to inject antibiotic at the induction of anaesthesia and to reduce the duration of their administration. PMID- 9750818 TI - [Latex allergy: how do you treat an emergency?]. PMID- 9750819 TI - [What is the correct position for the endotracheal probe prior to laparoscopic surgery?]. PMID- 9750820 TI - [Hepatitis C and anesthesia]. PMID- 9750821 TI - [Clinical applications of cisatracurium]. PMID- 9750822 TI - [Clinical aspects of resorption syndrome and therapeutic principles]. PMID- 9750823 TI - [Easy to handle fibroscope]. PMID- 9750825 TI - Medical technologies: concern about and assurance of value. PMID- 9750826 TI - Design and analysis of ROC studies: discriminating between diet and regular soft drinks. PMID- 9750827 TI - Applications of decision analysis. PMID- 9750828 TI - Practical issues in conducting clinical trials: randomized, controlled trials of imaging studies. PMID- 9750829 TI - Meta-analysis of diagnostic procedures: a brief overview. PMID- 9750830 TI - Measuring the effects of medical imaging on physicians' diagnostic and therapeutic thinking. PMID- 9750831 TI - Rationale for and measurement of patient preferences and health-related quality of life. PMID- 9750832 TI - Biases in the assessment of diagnostic innovations. PMID- 9750833 TI - Cost accounting for evaluating and planning of radiology resources. PMID- 9750834 TI - Subspecialty care in radiology: oncologic imaging. PMID- 9750835 TI - Measuring preferences for living in U.S. states: a comparison of the rating scale, time trade-off, and standard gamble. PMID- 9750836 TI - Decision analysis of cost-effectiveness of magnetic resonance angiography for mass screening for intracranial aneurysms. PMID- 9750837 TI - Cost-effectiveness of whole-body PET imaging in non-small cell lung cancer and malignant melanoma. PMID- 9750838 TI - Diagnostic imaging and the outcome of acute lower gastrointestinal bleeding. PMID- 9750839 TI - Measuring the effects of medical imaging in patients with possible cerebellopontine angle lesions: a four-center study. PMID- 9750840 TI - Digital mammography: a model for assessing cost-effectiveness. PMID- 9750841 TI - Government policy on medical radiology in Japan. PMID- 9750842 TI - Reorganization of diagnostic imaging in south Sweden: realization and cost effectiveness. PMID- 9750843 TI - Outcomes and benefits of CT-guided biopsy. PMID- 9750844 TI - Cost-effective diagnostic algorithms in pulmonary embolism: an updated analysis. PMID- 9750845 TI - Outcome of cervical-spine MRI. PMID- 9750846 TI - Symptoms or no symptoms: effectiveness of chest radiography. PMID- 9750847 TI - Cost evaluation of breast cancer screening in France. PMID- 9750848 TI - Conceptual framework for cost-effectiveness of imaging tests. PMID- 9750849 TI - Cost-effectiveness decision analysis of mass screening for lung cancer. PMID- 9750850 TI - Prediction of patient outcome after radical prostatectomy by imaging: choice of outcome measure. PMID- 9750851 TI - Quality assurance and clinical utility in radiology: a study of abdominal ultrasound. PMID- 9750852 TI - Standardizing cost-effectiveness analyses: the panel on cost-effectiveness in health and medicine. PMID- 9750853 TI - Quality of care in the USA in the 1990s: emphasis areas and stakeholders. PMID- 9750854 TI - The Iron Lady: reform and be damned--a cost and benefits analysis of UK radiology post reform. PMID- 9750855 TI - Implementing HCFA guidelines on appropriate use of nonionic contrast agents for diagnostic arteriography: effects on complication rates and management costs. PMID- 9750856 TI - Concurrent medicine: the next trend in the industrialization of health care. PMID- 9750857 TI - Reducing healthcare costs: how suppliers can contribute. PMID- 9750858 TI - Business solutions in radiology: a supplier's view. PMID- 9750859 TI - UCSF-Stanford health-care merger to create value for academic medicine. PMID- 9750860 TI - Health-care policy toward medical imaging experience in some European countries. PMID- 9750861 TI - The economy of health service in Russia and the development of modern diagnostic modalities. PMID- 9750862 TI - Cost-effectiveness contribution of contrast media: a discussion of perspectives. PMID- 9750863 TI - Technology assessment and reimbursement for future new imaging technologies. PMID- 9750865 TI - The Singapore system. PMID- 9750864 TI - Setting fees for medical services: a model for establishing a comprehensive medical service fees structure based on an integrated approach to service components cost analysis. PMID- 9750866 TI - Radiology and health-care policy in the Czech Republic. PMID- 9750867 TI - Health-care scenario in India. PMID- 9750868 TI - Current status and management of computed tomography and magnetic resonance imaging in China. PMID- 9750869 TI - Mobile Breast Care Center: a joint program of the Department of Defense, U.S. Public Health Service's Office on Women's Health, and the National Cancer Institute. PMID- 9750870 TI - Cost-effectiveness of digital radiography. PMID- 9750871 TI - Radiology research and the NIH: problems and future prospects. PMID- 9750872 TI - Academy of Radiology Research. PMID- 9750873 TI - Cost-effectiveness in radiology: perspective of the European Association of Radiology. PMID- 9750874 TI - Advancing the transition of imaging research into clinical practice. PMID- 9750875 TI - Cost-effective medical education in developing countries. PMID- 9750876 TI - Cost-effectiveness of image-guided surgery. PMID- 9750877 TI - The inevitability of 3D visualization: technology trends and cost/benefit. PMID- 9750878 TI - Radiology department management by radiologists. PMID- 9750879 TI - Economic impact of modern telematic technologies. PMID- 9750880 TI - An ISO-quality system in the radiology department: a benefit analysis. PMID- 9750881 TI - The new interventional center: experiences after 12 months of operation. PMID- 9750882 TI - Financial impact of Denmark's decision to restrict use of high-osmolar contrast media. PMID- 9750883 TI - Nationwide cost comparison of MR and self-referred cardiovascular procedures. PMID- 9750884 TI - The present and future of magnetic resonance imaging in an era of cost containment in Europe. PMID- 9750885 TI - Cost minimization by interventional magnetic resonance imaging: a businessman's perspective. PMID- 9750886 TI - Costs and benefits of radiology: meeting summary, perspective, and plans for the future. PMID- 9750887 TI - Complementary roles of generalists and specialists in academic radiology. PMID- 9750888 TI - Monte Carlo validation of a multireader method for receiver operating characteristic discrete rating data: factorial experimental design. AB - RATIONALE AND OBJECTIVES: The authors conducted a series of null-case Monte Carlo simulations to evaluate the Dorfman-Berbaum-Metz (DBM) method for comparing modalities with multireader receiver operating characteristic (ROC) discrete rating data. MATERIALS AND METHODS: Monte Carlo simulations were performed by using discrete ratings on fully crossed factorial designs with two modalities and three, five, and 10 hypothetical readers. The null hypothesis was true for all simulations. The population ROC areas, latent variable structures, case sample sizes, and normal/abnormal case sample ratios used in another study were used in these simulations. RESULTS: For equal allocation ratios and small (Az = 0.702) and moderate (Az = 0.855) ROC areas, the empirical type I error rate closely matched the nominal alpha level. For very large ROC areas (Az = 0.961), however, the empirical type I error rate was somewhat smaller than the nominal alpha level. This conservatism increased with decreasing case sample size and asymmetric normal/abnormal case allocation ratio. The empirical type I error rate was sometimes slightly larger than the nominal alpha level with many cases and few readers, where there was large residual, relatively small treatment-by-case interaction and relatively large treatment-by-reader interaction. CONCLUSION: The results suggest that the DBM method provides trustworthy alpha levels with discrete ratings when the ROC area is not too large and case and reader sample sizes are not too small. In other situations, the test tends to be somewhat conservative or slightly liberal. PMID- 9750889 TI - Perceptual skill, radiology expertise, and visual test performance with NINA and WALDO. AB - RATIONALE AND OBJECTIVES: The goal of this study was to determine if radiologists possess superior visual search and analysis skills compared with those of laypeople. MATERIALS AND METHODS: In two experiments, radiologists and laypeople searched one of two complex pictorial scenes for hidden targets. Eye position was recorded during the search. Two measures of performance were obtained: accuracy of detecting targets as measured by using alternative free response receiver operating characteristic analysis and visual search efficiency as measured by using eye position analysis. RESULTS: There were no statistically significant differences in detection performance between radiologists and laypeople for either of the search tasks. Radiologists took longer on average to search the images and to first fixate on the targets than did the laypeople. For both groups, true-positive and false-positive decisions were associated with longer dwell times than true-negative decisions. As with radiology search tasks, false negative decisions were also associated with longer dwell times than true negative decisions. CONCLUSION: Performance on two visual search and detection tasks indicate that radiologists do not possess superior visual skills compared with laypeople. Radiology expertise is more likely to be a combination of specific visual and cognitive skills derived from medical training and experience in detecting and determining the diagnostic importance of radiographic findings. PMID- 9750890 TI - Characteristics of regions suspicious for pulmonary nodules at chest radiography. AB - RATIONALE AND OBJECTIVES: This study was performed to determine physical characteristics of areas on chest radiographs that are suspicious but not definitive for the presence of a pulmonary nodule and the characteristics of areas that contain an obvious nodule. MATERIALS AND METHODS: Two groups of patients were identified: those who had an area at plain radiography that was suspicious for a pulmonary nodule and underwent fluoroscopy for further evaluation (138 patients, 142 areas) and those who had an obvious nodule at plain radiography who underwent computed tomography for further evaluation (72 patients, 97 areas). The measured characteristics of the region of interest included size, circularity, compactness, contrast, and location. RESULTS: A comparison of the data show that while there was some difference between these groups of patients with regard to location of the nodules, there were essentially no differences with regard to size, circularity, compactness, and contrast of the regions of interest. CONCLUSION: Size, circularity, compactness, contrast, and location are not sufficient to distinguish pulmonary nodules from other suspicious regions on the chest radiograph. PMID- 9750891 TI - In vitro function of an adjustable temporary venous spring filter: comparison with the temporary RF02 filter and the permanent Greenfield vena cava filter. AB - RATIONALE AND OBJECTIVES: The authors compared in vitro function of a temporary venous spring filter with that of a temporary RF02 filter and a permanent Greenfield filter. MATERIALS AND METHODS: All three types of filters were placed in thin polyethylene tubes (diameters, 10.0-18.0 mm). Physiologic saline was substituted for flowing blood, and blood clots of three sizes (6 x 10 mm, 6 x 20 mm, 9 x 20 mm) were funneled to the filters. Clot-trapping ability of each filter and elevation of intraluminal pressure after clot trapping were assessed for each tube size. RESULTS: No statistically significant elevation in intraluminal pressure was detected immediately after placement of any filter. The clot trapping ability of the spring filter and of Greenfield filter were slightly lower than that of the RF02 filter, but the differences were not statistically significant. After filters had trapped large clots, a high pressure gradient was detected in the 10.0-mm tube for all filters. The spring filter was associated with a higher pressure than the other filters in the 12.0-mm tube (P < .05). CONCLUSION: In vitro function of the spring filter was satisfactory in comparison with that of the RF02 filter and the Greenfield filter. For efficient filtering in the inferior vena cava, development of a larger version of the filter may be necessary. PMID- 9750892 TI - Graduate medical education financing: effect of the Balanced Budget Act of 1997. PMID- 9750893 TI - Update on long-term goals for medical image perception research. PMID- 9750894 TI - Role of magnetization transfer imaging in bone tumors. AB - RATIONALE AND OBJECTIVES: The authors evaluated the role of magnetization transfer imaging in differentiation of bone tumors, with special emphasis on cartilaginous tumors. MATERIALS AND METHODS: Fifty-one patients with skeletal tumors or tumor-like lesions who had undergone magnetic resonance (MR) imaging were included. The tumors were divided into three groups according to their gross appearance and the origin of tissue: cyst, cartilaginous tumor, and noncartilaginous solid tumor. A gradient-recalled acquisition in the steady state sequence was used for MR imaging, and examinations were performed both with and without off-resonance magnetization transfer pulses. Magnetization transfer ratios were obtained as an indicator of magnetization transfer effect of the lesions and were compared. Then, the magnetization transfer ratios of all tumors in the cartilaginous tumor group were compared. The magnetization transfer ratios of benign and malignant solid tumors were also compared. RESULTS: The mean magnetization transfer ratio for cartilaginous tumors was 0.31 +/- 0.08 (standard deviation), and that of cysts and noncartilaginous solid tumors was 0.07 +/- 0.03 and 0.40 +/- 0.14, respectively. Comparisons between the three groups showed statistically significant intergroup differences in magnetization transfer ratio (P < .05). In the cartilaginous tumor category, enchondroma and low-grade chondrosarcoma had a lower magnetization transfer effect than chondroblastoma and mesenchymal chondrosarcoma. The mean magnetization transfer ratios of benign (n = 28) and malignant (n = 18) tumors were 0.35 +/- 0.11 and 0.39 +/- 0.15, respectively; there was no statistically significant intergroup difference (P = .14). CONCLUSION: Magnetization transfer imaging could be useful for categorizing bone tumors as cysts, cartilaginous tumors, or noncartilaginous solid tumors. PMID- 9750895 TI - Imaging evaluation of chest wall tumors in children. PMID- 9750896 TI - Residents: be aggressive in your own education! PMID- 9750897 TI - Classification of dystonia. PMID- 9750898 TI - Physiology of dystonia. PMID- 9750899 TI - Pathophysiology of dystonia. PMID- 9750900 TI - Thalamic single neuron and electromyographic activities in patients with dystonia. PMID- 9750901 TI - Excitability of motor cortex in patients with dystonia. PMID- 9750902 TI - Opioid peptide precursor expression in animal models of dystonia secondary to dopamine-replacement therapy in Parkinson's disease. PMID- 9750903 TI - Characterization of a rodent model in which to investigate the molecular and cellular mechanisms underlying the pathophysiology of L-dopa-induced dyskinesia. PMID- 9750904 TI - Abnormal cerebellar output in the genetically dystonic rat. PMID- 9750905 TI - Clinical-genetic spectrum of primary dystonia. PMID- 9750906 TI - The gene (DYT1) for early-onset torsion dystonia encodes a novel protein related to the Clp protease/heat shock family. PMID- 9750908 TI - Familial cervical dystonia, head tremor, and scoliosis: a case report. PMID- 9750907 TI - The role of the DYT1 gene in secondary dystonia. PMID- 9750909 TI - The epidemiology of the primary dystonias in the north of England. PMID- 9750910 TI - Abnormal brain networks in DYT1 dystonia. PMID- 9750912 TI - Advanced neuroimaging methods in the study of movement disorders: dystonia and blepharospasm. PMID- 9750911 TI - Activation positron emission tomography scanning in dystonia. PMID- 9750913 TI - Decreased [18F]spiperone binding in putamen in dystonia. PMID- 9750914 TI - Medical therapy and botulinum toxin in dystonia. PMID- 9750915 TI - Surgical treatment of dystonia. PMID- 9750916 TI - Intrathecal baclofen in the treatment of dystonia. PMID- 9750917 TI - GPi pallidotomy for dystonia: clinical outcome and neuronal activity. PMID- 9750918 TI - Experience with stereotactics for dystonia: case examples. PMID- 9750919 TI - Botulinum toxin type B: an open-label, dose-escalation, safety and preliminary efficacy study in cervical dystonia patients. PMID- 9750920 TI - Dose standardization of botulinum toxin. PMID- 9750921 TI - Laryngeal dystonia (spasmodic dysphonia): observations of 901 patients and treatment with botulinum toxin. PMID- 9750922 TI - Dystonias responding to levodopa and failure in biopterin metabolism. PMID- 9750923 TI - Molecular and biochemical aspects of hereditary progressive and dopa-responsive dystonia. PMID- 9750924 TI - Atypical presentations of dopa-responsive dystonia. PMID- 9750925 TI - Tetrahydrobiopterin metabolism and GTP cyclohydrolase I mutations in L-dopa responsive dystonia. PMID- 9750926 TI - Defects of biopterin metabolism and biogenic amine biosynthesis: clinical diagnostic, and therapeutic aspects. PMID- 9750927 TI - Dopa-responsive dystonia: a syndrome of selective nigrostriatal dopamine deficiency. PMID- 9750929 TI - Inherited myoclonus-dystonia syndrome. PMID- 9750928 TI - The effect of GTP cyclohydrolase-1 on tyrosine hydroxylase expression: implications in DOPA-responsive dystonia. PMID- 9750930 TI - Rapid-onset dystonia-parkinsonism: a report of clinical, biochemical, and genetic studies in two families. PMID- 9750931 TI - Molecular genetic analysis of Lubag. PMID- 9750932 TI - The socioeconomic implications of dystonia. PMID- 9750934 TI - Sustainable production: a proposed strategy for the work environment. AB - BACKGROUND: In the future, competitive industries will need to design for environment, health and safety as well as for productivity. Although the new areas of pollution prevention and clean production have evolved to address the design of production processes with concerns for the ambient environment, current pollution prevention models do not include explicit concerns for health, safety, and the work environment. The field of occupational health and safety has much to contribute to improve current pollution prevention approaches and solutions. METHODS: The application of work environment disciplines will need to be expanded from the conventional focus on "end-of-pipe" assessment and solutions, which take the production processes and resulting hazards as a given, to include a new focus on materials selection and process redesign. To make this shift, a new framework called "sustainable production" is proposed. The basic unit of sustainable production is the production process. The framework integrates a focus on the ambient and work environment along with a focus on productivity and the economic viability of the business enterprise in setting production process design parameters. RESULTS: By shifting the focus of occupational and environmental health and safety from exposure control to process design, sustainable production reduces the likelihood that concerns for health, safety, and the environment will be seen as antagonistic to productivity and economic development. To move a firm toward sustainable production, occupational health and safety professionals will need to participate in interdisciplinary workplace teams that design and build new production processes and that continuously evaluate and redesign existing processes. CONCLUSIONS: This new strategy requires an expansion of the role of the occupational health and safety professional to include evaluation and redesign of processes that produce goods and services as well as the conventional evaluation of chemical, physical, and biological agents, work practices, and ergonomics. This expansion of occupational and environmental health and safety requires new research to develop the scientific and public policy basis of sustainable production. PMID- 9750935 TI - Prevalence of occupational lung disease in a random sample of former mineworkers, Libode District, Eastern Cape Province, South Africa. AB - BACKGROUND: Gold mineworkers in South Africa are exposed to high levels of silica dust as a result of which they are at risk of developing silicosis, which is a compensable disease. The incidence of tuberculosis is also high. METHODS: To determine the prevalence of occupational lung disease and the previous compensation history in former migrant mineworkers, a study was undertaken in a random sample of men living in Libode, a rural district of Eastern Cape Province, South Africa. Two hundred thirty-eight ex-mineworkers were examined according to a protocol that included chest radiography and spirometry. Chest radiographs were read into the International Labour Organisation (ILO) classification for pneumoconioses by two readers. RESULTS: The mean age was 52.8 years, and the mean length of service was 12.15 years. The prevalence of pneumoconiosis (> or = ILO 1/0) was 22% and 36% (variation by reader). For both readers, a significant association between length of service and pneumoconiosis and between pneumoconiosis and reduction in FVC and FEV was found. Twenty-four percent of study subjects were eligible for compensation. CONCLUSION: There is a high prevalence of previously undiagnosed, uncompensated pneumoconiosis in the study group. As a result of the failure to diagnose and compensate occupational lung disease, the social and economic burden of such disease is being borne by individuals, households, and the migrant labor-sending communities as a whole. PMID- 9750936 TI - Carcinogenic implications of the lack of tremolite in UICC reference chrysotile. AB - Using light and electron microscopy analysis, as well as electron diffraction, and energy-dispersive x-ray analysis, an aliquot of UICC chrysotile B was analyzed with special attention given to any tremolite contamination. Polarized light microscopy, with its limit of detection of approximately 1 micron when using dispersion staining, revealed chrysotile as the only fibrous asbestos component. Analytical electron microscopy at 333,000x of more than 20,000 consecutive fibers showed only the tubular morphology characteristic of chrysotile. These findings highlight that when this sample was used for exposure disease induced in animal models correlates with chrysotile-induced pathology, and does not support an explanation based on the "amphibole hypothesis." Thus, chrysotile should be considered as having the biologic ability to produce cancers, including mesotheliomas, based on the extensive use of this material as a standard reference material. PMID- 9750937 TI - Polymorphisms in the promoter of the tumor necrosis factor-alpha gene in coal miners. AB - Tumor necrosis factor-alpha (TNF) is recognized as a central mediator of mineral dust-induced lung fibrosis, and genetic polymorphisms of the TNF promoter have been reported to influence levels of TNF production. To assess whether polymorphisms within the TNF promoter gene are associated with susceptibility to coal workers' pneumoconiosis (CWP), the DNA of 78 coal miners was typed for G-to A transitions at positions -238 and -308. Our results show that frequency of A 308 genotype (T2) is significantly overpresented in coal miners with CWP (50%), as compared with miners without CWP (25%) and controls (29%). After correction for cumulative dust exposure and smoking, the A-308 transition genotype is still associated with the presence of CWP (OR = 3.0, 95% CI = 1.0-9.0). Both A-238 and A-308 transition genotypes were related to TNF release from endotoxin-stimulated blood monocytes; only the A-238 transition and not the A-308 transition was associated to coal dust-induced TNF release. In summary, this study shows that the A-308 transition is related to CWP, but this relation is not paralleled by a different TNF release in this genotype. A larger number of patients coupled to frequent TNF release are required to evaluate genotype screening to estimate individual health risks for effects of coal mine dust exposure. PMID- 9750938 TI - Malingering assessment in behavioral toxicology: what, why, and how. AB - Neurobehavioral assessment is frequently made in a forensic context. The cognitive assessment may be biased due to an international manipulation of data by the patient motivated by attainment of compensation, that is, malingering. Although malingering is highly relevant in behavioral toxicology, the issue and its assessment are underrepresented in the literature. A routine assessment of malingering is important to reduce false-positive and false-negative errors in assessment, thereby establishing the credibility and validity of behavioral assessment. In the long run, the routine inclusion of malingering measurements might reduce claims and encourage employers to be more cooperative in behavioral toxicology studies. Guidelines for malingering assessment and research, inferred from the clinical and research literature, are discussed. Sensitivity to the problematic issues involved in assessing malingering behavior is an important step toward malingering detection in the clinical setting and to the establishment of assessment methods that are less confounded by these issues. PMID- 9750939 TI - A multidimensional evaluation of fire fighter training for hazardous materials response: first results from the IAFF Program. AB - The International Association of Fire Fighters (IAFF) course on hazardous materials training for first responders is described together with an evaluation plan that includes multiple levels of assessment. Trainee appraisals of the course, shifts in their ratings of task competencies, gains in knowledge quiz scores, and self-reports on actions reflecting lessons learned from the course are among the measures used. Evaluations of courses given in several city fire departments found more than 60% of trainee judgments of course quality and utility to be highly favorable, along with significant post-course improvements in their competency ratings and quiz scores. Follow-up interviews with samples of trainees also suggested more self-protective behaviors and preventive actions being taken with regard to alarms and risks of hazardous materials exposures. However, cross-comparing the results for the various evaluation measures gave only limited support to a popular evaluation model that hypothesized that they would be interdependent. Limitations in appreciating technical course subjects, the value of add-on or refresher instruction, and variable risk experiences are noted in explaining differences in some training results. PMID- 9750940 TI - Improving safety for teens working in the retail trade sector: opportunities and obstacles. AB - BACKGROUND: Using both quantitative and qualitative data, this study examined teen workers' perceptions about their work environments and the ways in which teens believe workplaces can be made safer. METHODS: We conducted telephone interviews (n = 117) and six focus groups (n = 49) with two separate samples of North Carolina teens who worked in the retail trade sector. RESULTS: Survey findings indicate one-fifth of teens used equipment they thought dangerous; nearly 40% always or often felt rushed at work; and about half received training on how to avoid injury. Teens in the focus groups expressed concerns about workplace physical hazards, the threat of assault, being rushed, and having little power in the work environment. They also indicated that their workplace safety training was ineffective and that child labor laws were unnecessary. CONCLUSIONS: In order to be effective, interventions targeted at working teens need to address the organization of work and adolescent-manager interaction patterns. PMID- 9750941 TI - Acute traumatic injuries in automotive manufacturing. AB - Motor vehicle manufacturing, with its varied tasks, challenging work environment, and diverse worker populations, presents many hazards to employees. This study examined routinely collected surveillance data from a major motor vehicle manufacturer to identify injury types, high-risk workers, causes of injury, and factors associated with work loss. Injury and personnel data were used to calculate injury rates. Injury data were from the routinely collected medical and safety surveillance system on occupational injuries. The number of persons working in the plants was estimated using year-end personnel reports. Key word searches supplementing the analyses provided insight into the specific circumstances of injury. The most common injuries were sprains/strains (39% of the total), lacerations (22%), and contusions (15%). Forty-nine percent of the injuries resulted in one or more lost or restricted workdays; 25% resulted in 7 or more lost or restricted workdays. The injuries most likely to result in work loss were amputations, hernias and fractures. Sprains/strains accounted for 65% of all lost workdays. Injury rates ranged from 13.8 per 100 person-years at stamping plants to 28.7 at parts depots. Even within similar types of plants, injury rates varied widely, with a twofold difference among the individual assembly plants in overall injury rates. Injury surveillance systems with descriptive data on injury events shed light on the circumstances under which certain types of injuries occur and can provide the basis for preventive interventions. Sources of variation and potential biases are discussed, providing guidance for those interested in designing and using surveillance systems for occupational injuries. PMID- 9750942 TI - An evaluation of the prevalence of latex sensitivity among atopic and non-atopic intensive care workers. AB - BACKGROUND: Exposure to latex is known to cause an array of symptoms, including pruritus, dermatitis, erythema, and urticaria. Workers at elevated risk for latex exposure include health care personnel whose repeated patient contact or surgical work require extensive use of latex gloves. This study evaluated the prevalence of latex allergies in atopic and non-atopic intensive care workers and sought to determine the impact of risk factors such as frequency of glove use and hand washing on latex sensitization. METHODS: We evaluated the prevalence of latex sensitivity in 122 intensive care unit (ICU) workers using a questionnaire and skin prick test. Atopy and latex sensitivity were determined by skin prick test using a battery of common inhalant allergens and an extract prepared from the gloves used in the ICU. Frequency of glove use and hand washing were determined by questionnaire. RESULTS AND CONCLUSIONS: Forty ICU workers (32.8%) were considered atopic by having at least one positive response to the inhalant allergens. Atopic ICU workers were more likely to have positive latex skin test than non-atopic ICU workers (atopic vs. non-atopic workers: p < 0.001, odds ratio = 14.2). Frequency of current glove use or hand washing frequency were not significant predictors of a positive response to latex; however, a positive history of atopic eczema and family history of allergies, as determined by questionnaire were significant predictors of a positive response to latex antigens. PMID- 9750943 TI - Suicide mortality and pesticide use among Canadian farmers. AB - BACKGROUND: An exploratory, case-control study was used to investigate a new hypothesis about suicide among farm operators. This hypothesis suggested a biologically plausible link between exposures to certain pesticides and the occurrence of suicide among farm operators. These analyses were based on data from the Canadian Farm Operator Cohort. METHODS: Canadian male farm operators who committed suicide between 1971-1987 (n = 1,457) were compared with a frequency matched (by age and province) sample of control farm operators (n = 11,656) who were alive at the time of death of individual cases. Comparisons focused on past exposures to pesticides reported to the 1971 Canada Census of Agriculture. RESULTS: Multivariate logistic regression analyses indicated no associations between suicide and (1) acres sprayed with herbicides, (2) acres sprayed with insecticides, and (3) the costs of agricultural chemicals purchased; after controlling for important covariates. There was, however, a suggestive increase in risk for suicide associated with herbicide and insecticide spraying among a subgroup of farm operators who were most likely to be directly exposed to pesticides: OR = 1.71 (95% CI = 1.08-2.71) for 1-48 vs. 0 acres sprayed. Additional risk factors that were identified included seasonal vs. year-round farm work (OR = 1.68; 95% CI = 1.15-2.46); and high levels of paid labor on the farm (e.g., OR = 1.61; 95% CI = 1.24-2.10, for > 13 vs. 0 weeks per year). Factors that were protective included marriage (odds ratio (OR) = 0.69; 95% confidence interval (CI) = 0.58-0.81), having more than one person resident in the farm house (e.g., two vs. one person; OR = 0.62; 95% CI = 0.42-0.92); and higher levels of education (e.g., postsecondary vs. primary; OR = 0.40; 95% CI = 0.17-0.96). CONCLUSIONS: This study does not provide strong support for the main hypothesis under study, that exposure to pesticides is an important risk factor for suicide among farmers. Although secondary to the main hypothesis, a number of other risk factors for suicide were suggested. These have implications for the future study and targeting of suicide prevention programs in rural Canada. PMID- 9750944 TI - Relative chronic effects of different occupational dusts on respiratory indices and health of workers in three Ethiopian factories. AB - The respiratory effects of dusts in different sections of yarn, cement, and cigarette factories were studied in 211 nonsmoking male and female workers aged 21-57 years. The controls used were 211 healthy nonsmoking and nonexposed male and female subjects aged 20-57 years from the general population. Forced vital capacity (FVC), forced expiratory volume in 1 sec (FEV1), FEV1/FVC ratio, forced expiratory flow (FEF200-1,200 ml), forced mid-expiratory flow (FMF25-75%) and peak expiratory flow rate (PEFR) were recorded in all subjects with and without respiratory symptoms. Taking exposures to all dusts of different concentrations together, it was found that the frequency of respiratory illness was greater among exposed workers (40.5% in males, 36% in females) than it was among controls (21.6% in males, 18% in females). In exposed subjects, the symptom prevalence was only 4.5% higher in males than in females. The mean lung function indices, including FEV1, FEV1%, FEF200-1,200 ml, FMF25-75%, and PEFR, in subjects exposed to all dusts in general decreased markedly, with dust concentration being more important than duration of exposure, and FMF being affected slightly more consistently. About 38.4% of the dust-exposed subjects developed corresponding respiratory illnesses including chronic cough (24.7%), chronic bronchitis (21.8%) and bronchial asthma (24.2%). The respective control values were 9.0%, 9.5%, and 8.5%. Exposure to different occupational dusts resulted in the development of respiratory illness with different rates of prevalence. The effects of exposure to cotton and cement dusts on respiratory health of exposed subjects were relatively more significant (p < 0.001) than that of exposure to tobacco dust (p < 0.05). PMID- 9750946 TI - Hypersensitivity pneumonitis in a machinist. AB - Hypersensitivity pneumonitis (HP), or extrinsic allergic alveolitis, is a patchy, interstitial lung disease that involves an immunologic reaction of the lung to repeated inhalation of foreign antigens. In this report, we describe a machinist with exposure to metalworking fluids (MWFs) and biopsy-confirmed HP. Return to work, which could be equated with a retrospective workplace-specific bronchoprovocation test, proved that working within an environment in which MWFs were used was associated with clinical deterioration in the patient's pulmonary status and with clinical improvement after removal from exposure. PMID- 9750945 TI - Lung health among boilermakers in Edmonton, Alberta. AB - BACKGROUND: Construction boilermakers may be exposed to a variety of substances, including asbestos and welding fumes. Past studies of boilermakers have shown increases in mortality from lung cancer and asbestosis and radiographic changes consistent with asbestos exposure. METHODS: Respiratory symptoms, lung function, and radiographic changes were compared for 102 actively employed boilermakers with 20 or more years of union membership and 100 telephone workers. Posteroanterior chest radiographs were evaluated by two experienced chest physicians, with a third arbitrating disagreed films. Union members were further categorized as boilermakers (n = 50) or welders (n = 52), based on longest service. Lung health was also compared with employment in a number of work sectors for time, and time-weighted exposure to dust and fumes. RESULTS: Boilermakers had more respiratory symptoms than telephone workers, but lung function did not differ. Radiographic changes were more common among the boilermakers (20% with any change, 8% circumscribed, and 9% diffuse pleural thickening). None of the boilermakers had small radiographic opacities. Several symptoms suggestive of bronchial responsiveness were associated with fume exposures in the gas and oil industry. Workers whose longest service was as a boilermaker demonstrated more symptoms than did welders. FEV1, FEV1/FVC, FEF25 75, and FEF50 were significantly lower among boilermakers compared with welders. CONCLUSION: Health screening programs for these workers are warranted. PMID- 9750947 TI - A nested case-control study of bladder cancer incidence in a steel manufacturing plant. AB - BACKGROUND: A surveillance study of bladder cancer incidence in northwestern Illinois detected a number of cases who had worked in a large steel manufacturing plant. To investigate these cancers further, a nested case-control study of bladder cancer was conducted at this plant. METHODS: Cases from the surveillance study were matched to company personnel files. Four controls per case were selected from company files and frequency matched on age (cases and controls were all white males). Employment histories were abstracted from company records and odds ratios calculated according to job titles and departments worked. Cases and controls who held only white collar jobs were excluded from the analysis. RESULTS: Results for 16 cases and 75 controls showed elevated odds ratios for heaters (OR = 21.1, 95% cCI = 2.2-205.8) based on three cases. Heaters monitored electric reheat furnaces and may have been exposed to polycyclic aromatic hydrocarbons. Previous studies have shown moderate risks for bladder cancer among furnace operators, but most of these studies were also based on a small number of cases. CONCLUSIONS: Additional studies of furnace operators' exposures and cancer risks are needed to investigate these results further. PMID- 9750948 TI - Corruption of the medical literature: a second visit. PMID- 9750949 TI - Re: An early study of pulmonary asbestosis among manufacturing workers: original data and reconstruction of the 1932 cohort. PMID- 9750950 TI - 'Dust and mirrors' or 'corruption is in the eye of the beholder'. PMID- 9750951 TI - Restoration of nitric oxide function in human hyperlipidaemia, congestive heart failure and liver cirrhosis. AB - 1. There is accumulating evidence for a range of abnormalities in the nitric oxide (NO) signalling cascade in human cardiovascular disorders. 2. In the present review we assess the literature detailing such evidence in early (hyperlipidaemia) and end-stage (heart failure) disease, with emphasis on the mechanisms by which the disturbances are thought to occur. 3. Strategies for the correction of disturbed NO signalling in these states are reviewed and include both prescribed pharmacological interventions, such as lipid-lowering therapy and novel uses of angiotensin-converting enzyme inhibitors, as well as non pharmacological interventions, such as exercise and dietary supplementation with L-arginine and n-3 polyunsaturated fatty acids. 4. In addition to a decreased production/function of NO, the possible detrimental effects of a chronic elevation in NO production in patients with liver cirrhosis, together with a novel use of antibiotics to correct this perturbation, is outlined. PMID- 9750953 TI - Role of the mesolimbic dopamine system in cardiovascular homeostasis. Stimulation of the ventral tegmental area modulates the effect of vasopressin on blood pressure in conscious rats. AB - 1. The possible role of the ventral tegmental area (VTA) and its dopaminergic projections in cardiovascular regulation is reviewed. 2. Our own work has shown that stimulation of the VTA by local microinjection of the substance P analogue DiMe-C7 caused an increase in blood pressure. The mechanism of the pressor response was an interaction of central dopaminergic activation, most likely at the level of the baroreflex, with the circulatory actions of vasopressin. 3. These findings are important for a possible role of the mesolimbic dopamine system in cardiovascular homeostasis. Several studies reviewed here show that neuronal activity of the VTA and its mesolimbic projections is altered by changes in blood pressure, salt and electrolyte balance, stress and food and water intake. 4. The VTA and mesolimbic dopamine system, while playing a widely accepted role in locomotor activity, cognition and reward mechanisms, may also be involved in the integration of sensory and behavioural information with cardiovascular homeostasis. PMID- 9750952 TI - Amylin: physiological roles in the kidney and a hypothesis for its role in hypertension. AB - 1. There are high-affinity binding sites for amylin in the renal cortex associated with proximal tubules. These appear to represent seven transmembrane (heptatopic) receptors that are known to form ternary complexes with G-proteins and activate second messenger systems. 2. Amylin stimulates sodium/water reabsorption from the basolateral side of the proximal tubules and plays a role in sodium homeostasis. 3. The transient expression of amylin-like mRNA has been detected perinatally, using in situ hybridization, in the subnephrogenic zone of the metanephros and is associated with proximal tubules of the developing nephron. There it is thought to play a role as a growth factor for brush border epithelial cells in the developing kidney and in renal regrowth in the adult kidney. 4. In two models of hypertension, the spontaneously hypertensive rat (SHR) and one created surgically by subtotal nephrectomy, renal amylin receptors are activated. In the SHR, activation precedes the rise in blood pressure and suggests that activation of the amylin system may be an important event in the development of hypertension. PMID- 9750954 TI - Dopamine releases endothelium-derived relaxing factor via alpha 2-adrenoceptors in canine vessels: comparisons between femoral arteries and veins. AB - 1. We investigated the role of vascular smooth muscle alpha-adrenoceptor subtypes in the vasoconstrictor response of femoral arteries and veins to dopamine and whether the vasoconstriction is modified by endothelium-dependent relaxation mediated via the activation of alpha 2-adrenoceptors in ring preparations of femoral arteries and veins from mongrel dogs. 2. Dopamine contracted both arteries and veins in a dose-dependent manner and this contraction was inhibited by pretreatment with phentolamine or prazosin. Pretreatment with yohimbine shifted the dose-response curve for dopamine to the right in femoral veins, but not in arteries. 3. Phenylephrine contracted femoral arteries and veins in a dose dependent manner and this contraction was inhibited by pretreatment with prazosin. 4. Clonidine produced a bell-shaped dose-response curve in femoral veins and this curve was shifted upwards by pretreatment with NG-nitro-L-arginine (L-NNA). In contrast, femoral arteries were not affected by clonidine. NG-Nitro-L arginine potentiated contractile responses to dopamine in both veins and arteries. This potentiation was inhibited by yohimbine or by the removal of the endothelium in both arteries and veins. 5. These results suggest that dopamine contracts femoral arteries via stimulation of alpha 1-adrenoceptors and contracts femoral veins via stimulation of both alpha 1- and alpha 2-adrenoceptors and that these contractions are attenuated by the vasodilator action of dopamine via alpha 2-adrenoceptor-mediated release of endothelium-derived relaxing factor. PMID- 9750956 TI - Functional assessment of alpha 1-adrenoceptor subtypes in porcine coronary artery. AB - 1. alpha 1-Adrenoceptors are known to play an important role in vasoconstriction in response to adrenergic stimulation. However, the functional importance of alpha 1-adrenoceptor subtypes at the epicardial coronary artery remains unclear. We examined alpha 1-adrenoceptor subtypes by comparing functional affinities for alpha-adrenoceptor antagonists on noradrenaline (NA)-induced vasoconstriction in porcine denuded right coronary arteries. 2. Noradrenaline induced a dose dependent vasoconstriction in incubated vessel rings. Prazosin and phentolamine were potent and competitive antagonists for NA-induced contraction (pA2 10.27 and 9.03, respectively). In contrast, the selective alpha 2-adrenoceptor antagonist yohimbine had a low affinity (pA2 6.13). Two selective alpha 1A-adrenoceptor antagonists, WB 4101 and 5-methyl urapidil, were potent and competitive antagonists of alpha 1-adrenoceptor-induced contraction (pA2 10.67 and 8.90, respectively) and the selective alpha 1D-adrenoceptor antagonist BMY 7378 had a low affinity (pA2 6.06). Noradrenaline-induced contraction was insensitive to the alkylating effects of chlorethylclonidine. These observations indicate that the vasoconstriction is predominantly mediated by the alpha 1A-adrenoceptor subtype. This was also supported by a good correlation between pA2 values from the present study and reported binding affinities (pKi) of various alpha-adrenoceptor antagonists with cloned human alpha 1A-adrenoceptors (r = 0.98), but not for alpha 1B- or alpha 1D-adrenoceptor subtypes (r = 0.77 and 0.41, respectively). 3. Our results indicate that the alpha 1A-adrenoceptor is the main functional receptor subtype in porcine denuded coronary arteries. PMID- 9750955 TI - Age-related changes in aortic sensitivity to noradrenaline and acetylcholine in rats. AB - 1. The purpose of the present study was to determine the relationship between plasma and tissue lipid levels and the effects of age on vascular responses to noradrenaline (NA) and acetylcholine (ACh). 2. Studies were performed in young and aged rats and the response of endothelium-intact and -denuded aortic rings to NA and to ACh was measured. The plasma concentration of cholesterol (total, high density lipoprotein (HDL) and low-density lipoprotein (LDL)) and 17 beta oestradiol was determined, as was the aortic tissue content of phospholipids, cGMP and cholesterol (total, free and esterified). 3. Levels of all types of cholesterol in plasma and aorta increased with age; cholesterol levels in plasma correlated with those in the aorta; levels of phospholipid in the aorta did not increase with age but correlated with those of LDL cholesterol in plasma; levels of 17 beta-oestradiol did not change, but those of cGMP increased with age. 4. In endothelium-intact rings, the maximum tension developed by exposure to NA did not change, but the EC50 of NA increased with age and correlated with total cholesterol in the plasma and with the levels of all types of cholesterol in the aorta. In rings precontracted with NA, age decreased the maximum relaxation induced by ACh. The EC50 of ACh decreased with age and was inversely correlated with levels of cholesterol in the plasma and aorta. Treatment with NA increased cGMP levels in aged rats. Removal of the endothelium abolished the response to ACh and heightened the sensitivity to NA in young and aged rats. 5. Aortic endothelial cells seem to inhibit amine-induced contraction, while age-related changes in the levels of cholesterol in aortic tissue affect the sensitivity of the tissue to NA and ACh. PMID- 9750957 TI - Mobilization of lead by calcium versenate and dimercaptosuccinate in the rat. AB - 1. Calcium disodium ethylenediaminetetraacetate (CaNa2 EDTA) and meso-2,3 dimercaptosuccinic acid (DMSA) individually and in permutation-combination in various doses (0.1, 0.2 and 0.4 mmol/kg bodyweight) were investigated for their efficacy to mobilize lead from vital tissues into urine and faeces and to restore the lead-sensitive biochemical parameters in lead pre-exposed rats with a view to develop the most acceptable treatment regimen for lead poisoning with a minimal loss of endogenous essential elements. 2. The combined therapy was more effective than a single chelator treatment. 3. The combination of 0.2 mmol/kg CaNa2EDTA + 0.4 mmol/kg DMSA caused a lower depletion of zinc, calcium and iron but possessed almost equal capability to that of 0.4 mmol/kg CaNa2EDTA + 0.4 mmol/kg DMSA to produce urinary as well as faecal excretion of lead, to reduce the tissue burden of lead, including that of the brain, and to reverse lead-induced biochemical alterations. 4. The combination of 0.2 mmol/kg CaNa2EDTA + 0.4 mmol/kg DMSA has shown a definite improvement over previously reported combinations in terms of removal of lead from tissues, particularly the brain, restoration of urinary delta-aminolevulinic acid levels and a decrease in the loss of body zinc and is, therefore, recommended for the treatment of lead intoxication. PMID- 9750958 TI - Effects of nitric oxide synthase inhibition and endothelin ETA receptor blockade on haemodynamics in hypertensive rats. AB - 1. The objectives of the present study were to study regional differences in haemodynamics between spontaneously hypertensive (SHR) and normotensive Wistar Kyoto (WKY) rats induced by the nitric oxide synthase (NOS) inhibitor NG monomethyl-L-arginine (L-NMMA) and the endothelin ETA receptor antagonist BQ 123 in vivo in tissues known to be important for blood pressure (BP) regulation (heart, kidney and skeletal muscle). Furthermore, the effect of acetylcholine (ACh) infusion (2 micrograms/kg per min) was examined after L-NMMA or BQ 123. The microsphere method was used for determinations of cardiac index (CI) and regional haemodynamics. 2. NG-Monomethyl-L-arginine (20 mg/kg) increased BP (26-48%; P < 0.01) and reduced CI in both rat strains. BQ 123 (1 mg/kg) reduced BP slightly ( 4 to 11%; P < 0.05). 3. NG-Monomethyl-L-arginine significantly increased myocardial and skeletal muscle vascular resistance in SHR only; however, in the kidney, L-NMMA reduced blood flow and increased vascular resistance in both rat strains. 4. BQ 123 induced minor changes in regional haemodynamics that were not significantly different between the two strains. 5. Acetylcholine following BQ 123 induced an increase in myocardial blood flow in WKY rats, but decreased blood flow in SHR. Acetylcholine following L-NMMA reduced myocardial blood flow in both strains. 6. Acetylcholine following BQ 123 induced renal vasodilation in WKY rats but, following L-NMMA, ACh did not induce renal vasodilation in either rat strain. In contrast, L-NMMA did not abolish the vasodilation of acetylcholine in skeletal muscle in WKY rats. 7. In conclusion, the contribution of nitric oxide to basal vessel tone was not impaired in the heart, skeletal muscle and kidney in SHR. Antagonism of ETA receptors caused similar haemodynamic responses in both rat strains in these organs. Furthermore, NOS inhibition, but not ETA blockade, blunted the expected ACh-induced vasodilation in the heart and kidney in WKY rats, but not in skeletal muscle in both strains. PMID- 9750959 TI - Anti-proliferative effects of unmodified antisense oligodeoxynucleotides targeted against c-raf mRNA: use of poly (lysine/serine) copolymers or cationic lipopolyamines. AB - 1. It is now known that nuclease-resistant phosphorothioate antisense oligodeoxynucleotides (ODN) have some actions that are unrelated to antisense mechanisms. In the present study we assessed the anti-proliferative effects of phosphorothioate (PS) and phosphodiester (PO; unmodified) antisense ODN targeted against c-raf mRNA on pancreatic cancer cells in vitro, using poly (lysine/serine) copolymers conjugated with polyethylene glycol (PLSP) or cationic lipopolyamines (Transfectam) as carriers. 2. The anti-proliferative effect of the PO antisense ODN was significantly (P < 0.05) greater than that of the PS ODN, either complexed with PLSP (2 mumol/L ODN) or the Transfectam (0.5 mumol/L ODN). However, the effect of the PS or PO antisense ODN was not dependent on the antisense sequence. The c-raf mRNA levels, assessed by reverse transcription polymerase chain reaction, were obviously reduced by both PO and PS antisense ODN compared with mismatched ODN when complexed with the Transfectam (1 mumol/L ODN). 3. Although the anti-proliferative effects were mainly unrelated to antisense mechanisms, unmodified antisense ODN complexed with some carriers could be used as anti-tumour agents considering that synthetic carriers can be modified to improve functions, such as delivery. PMID- 9750960 TI - Autacoid interactions in the regulation of blood flow in the human placenta. AB - 1. Interactions between autacoids may play important roles in the regulation of blood flow in the foetal placenta. In order to investigate this aspect of placental haemodynamics, human normal-term placentae were perfused in vitro and the responses of the foetal vessels to various combinations of vasoactive agents were determined. 2. Vasoconstriction responses to 5-hydroxytryptamine (5-HT) were potentiated in the presence of endothelin-1 (ET-1), the thromboxane A2-mimetic U46619 and a nitric oxide synthase inhibitor, N-nitro-L-arginine (NOLA), but not in the presence of angiotensin II. 3. N-Nitro-L-arginine caused vasoconstriction of the perfused placenta and indomethacin attenuated this effect and blocked the potentiation of the 5-HT response by NOLA. 4. Indomethacin did not affect ET-1 induced pressure increases and infusion of U46619 had no effect on release of ET like immunoreactivity into the foetal placental circulation. 5. The present study provides evidence of interactions between several autacoids in human perfused placentae in vitro. These interactions may play important roles in foetal placental haemodynamics in normal or pathological situations. PMID- 9750961 TI - Metabolic effects of thioctic acid in rodent models of insulin resistance and diabetes. AB - 1. The antioxidant thioctic acid (TA) has been used in the treatment of diabetic neuropathy and recent studies have suggested that TA also has pancreatic and peripheral effects that improve glucose transport and metabolism. In the present study, the metabolic effects of TA were evaluated in rodent models of insulin resistance (fructose-fed Sprague-Dawley rat) and insulin deficiency (streptozotocin (STZ)-induced diabetic rat). Oral and intravenous glucose tolerance tests (OGTT and IVGTT, respectively) were performed in conscious rats after treatment with 50 mg/kg per day TA or vehicle for 5 days. 2. Fructose feeding for 7 days induced insulin resistance and impaired glucose tolerance and hypertriglycerideaemia. Treatment of fructose-fed rats with TA had no significant effect on fasting or stimulated glucose levels or on fasting triglyceride concentrations (e.g. the area under the curve for glucose (AUCglu) following OGTT was 1233 +/- 67 and 1284 +/- 59 in fructose-fed rats treated with either TA (n = 12) or vehicle (n = 12), respectively). Similarly, TA had no significant effect on IVGTT profiles in fructose-induced insulin resistance. 3. Low-dose STZ (80 mg/kg, i.p., over 2 days) induced hyperglycaemia, but TA had no significant glucose-lowering effects in STZ-diabetic rats (AUCglu (OGTT) following oral administration was 5507 +/- 27 and 5450 +/- 27 in TA (n = 12) and vehicle-treated (n = 12) rats, respectively). Nor did pretreatment with TA affect the diabetogenic response to STZ. 4. In contrast with previous in vitro studies reporting favourable metabolic effects of TA, the present study shows that after short-term oral therapy there are no significant improvements in glucose tolerance in rodent models of insulin resistance and insulin deficiency. Thioctic acid is unlikely to be of therapeutic benefit as an anti-diabetic drug in clinical practice. PMID- 9750962 TI - Evolution of surface activity related functions of vertebrate pulmonary surfactant. AB - 1. Pulmonary surfactant is a mixture of lipids and proteins that lines the air liquid interface of the lungs of all vertebrates. In mammals, it functions to reduce and vary surface tension, which helps to decrease the work of breathing, provide alveolar stability and prevent alveolar oedema. The present review examines the evolution and relative importance of these surface activity related functions in the lungs of vertebrates. 2. The surface activity of surfactant from fish, amphibians, birds and most reptiles is generally very low, correlating with a low body temperature and a low disaturated phosholipid content of their surfactant. In contrast, the surfactant of those reptiles with a higher preferred body temperature, as well as that of birds and mammals, has a much higher surface activity. 3. The two main functions of surfactant in mammals are to provide alveolar stability and to increase compliance of the relatively stiff bronchoalveolar lung. As the respiratory units of most non-mammalian vertebrates are up to 1000-fold larger and up to 100-fold more compliant, surfactant is not required for these functions. 4. In non-mammals, surfactant appears to act as an anti-glue preventing the adhesion of respiratory surfaces that may occur when the lungs collapse (e.g. during diving, swallowing of prey or on expiration). Surfactant also controls lung fluid balance. These functions can be fulfilled by a surfactant with relatively low surface activity and may represent the primitive functions of surface active material in vertebrate lungs. PMID- 9750963 TI - Urea: diverse functions of a 'waste' product. AB - 1. The urea cycle is essentially the simultaneous operation of two linear pathways, both primitive and widespread among animals; one is for arginine synthesis and the other is for arginine degradation to ornithine and urea. 2. All animals may have the genetic capacity to express a urea cycle and many diverse groups of animals, from flatworms to mammals, have a functional urea cycle. 3. Evolutionary changes in vertebrates of carbamylphosphate synthetase (CPS) are directed from glutamine-dependent (CPSIII) towards NH3-dependent (CPSI) ureagenesis. Invertebrates, cartilagenous fish and the coelacanth have CPSIII (i.e. glutamine-dependent), whereas lungfish, amphibians and amniote vertebrates have CPSI; the teleost Heteropneustes has CPSI-like activity. That the coelacanth has CPSIII and Heteropneustes has 'CPSI' suggests that the form of CPS may by physiologically related (CPSIII in a balancing solute role and CPSI in a terrestrial, air-breathing excretion role) rather than being phylogenetically constrained. 4. Urea is a major balancing osmolyte in marine cartilagenous fish, the coelacanth and a few amphibians and some aestivating terrestrial amphibians. It is a storage osmolyte in cocoon-forming aestivating lungfish and amphibians. 5. Urea contributes towards positive buoyancy in marine cartilagenous fish. 6. Urea functions for non-toxic N transport in ruminant and pseudoruminant mammals. 7. Urea is a major solute in the mammalian (but not avian) kidney, contributing to a renal medullary osmotic gradient; it is substantially reabsorbed by mammalian nephrons. 8. Urea is used as a preferred nitrogenous waste compared with ammonia at high ambient pNH3 or pH, with water restriction, or air breathing. 9. Urea synthesis maintains acid-base balance by the 1:1 stoichiometry of removal of HCO3- and NH4+. PMID- 9750965 TI - Are extant lungfish neotenic? AB - 1. The thyroid axis in developing lungfish is being explored to ascertain whether it shows similar deficiencies to those characteristic of neoteny in urodele amphibians. 2. At hatching, the pituitary of Neoceratodus forsteri comprises a lumen surrounded by a single layer of epithelial cells lying immediately below the hypothalamus, but unconnected to it. Over the first year of development, the number of pituitary cells increases and several cell types, including thyrotropes, can be recognized immunocytochemically, but the pituitary remains unconnected to the hypothalamus. 3. By treating the lungfish with thyroid inhibitor, no increase in thyroid uptake of iodine, indicative of induced pituitary thyroid-stimulating hormone (TSH) activity, could be demonstrated; neither was there any response to exogenous human TSH. In the liver, thyroid hormone receptors were found to be primarily of the alpha type. 4. Taken together, these findings suggest that up to 1 year of age, lungfish development is equivalent to amphibian premetamorphosis, which is consistent with neoteny but cannot be taken as evidence for neoteny unless confirmed at later stages of lungfish life history, which are yet to be studied. PMID- 9750964 TI - Evolution of thyroid hormone distribution. AB - 1. Appropriate distribution of thyroxine between the lipid-soluble compartments of cells and tissues and the extracellular aqueous space is established by binding to extracellular proteins. Among these proteins, transthyretin is of particular interest because it is the only one synthesized in the brain. 2. The evolutionary onset of transthyretin synthesis in cells of the blood-brain barrier precedes that in the liver, with the exception of a very short period of transthyretin synthesis in the liver of tadpoles, just prior to the climax of metamorphosis. In adult liver, transthyretin is only synthesized in endothermic vertebrates. 3. The affinity of transthyretin for thyroxine increases and that for 3,5,3'-triiodothyronine decreases during the evolution of eutherians from reptile/bird-like common ancestors. 4. A systematic change of the N-terminal region of transthyretin occurred during evolution, leading to shorter and more hydrophilic transthyretin N termini in eutherians compared with those in reptiles and birds. 5. The molecular mechanism of the evolution of the transthyretin N termini is a stepwise shift of the splice site at the intron 1/exon 2 border in the 3' direction. The most probable cause for this shift is a series of single base mutations. 6. As the N termini are located on the surface of transthyretin near the entrance to its central channel leading to the thyroxine binding sites, it is possible that a change in the structure of this region could influence the access of thyroxine to the binding sites. The increase in affinity for thyroxine could then be a driving force in the natural selection during evolution of transthyretins with shorter and more hydrophilic N termini. PMID- 9750966 TI - Evolution of daily torpor and hibernation in birds and mammals: importance of body size. AB - 1. The evolution of hibernation and daily torpor in mammals and birds remains a controversial subject. The original view was that use of torpor reflects a primitive thermoregulation, as it occurs in ancestral groups of mammals. 2. This view is no longer widely supported. However, the interpretation of a polyphyletic derivation of torpor also has been challenged because of the astonishing similarity of torpor patterns among various orders and even the two classes. 3. A recent argument is that mutations required for torpor and hibernation are unlikely to occur simultaneously and that torpor must be plesiomorphic (ancestral), although it is not functionally primitive. Homeothermy is interpreted as a loss of the ability to enter torpor in those groups that could survive without the requirement of heterothermic periods for energy conservation. 4. Interestingly, while torpor in mammals occurs in the phylogenetically old groups, lending support to the hypothesis of an ancestral derivation of torpor, the opposite is the case for birds. Modern bird groups and ancestral mammal groups contain mainly small species that often rely on fluctuating food supply, whereas modern mammalian orders and ancient bird orders contain the largest species with low energy requirements for maintenance and thermoregulation. 5. It is, therefore, possible that not phylogenetic position but size and diet determine the occurrence of heterothermy. Moreover, because endothermy and torpor in birds has apparently evolved separately from that in mammals and because it is possible that daily torpor and hibernation represent two distinct torpor patterns that evolved separately, a convergent evolution of torpor in endotherms cannot be excluded. PMID- 9750967 TI - Embryonic globins of the marsupial the Tammar Wallaby (Macropus eugenii): bird like and mammal like. AB - 1. Tammar Wallaby embryonic blood has been shown to have three alpha-like and two beta-like globin chains in its four haemoglobin components and partial sequences of several chains have been determined. 2. The major embryonic beta-like chain (epsilon) is similar to other mammalian embryonic beta-like chains on the basis of sequencing its first 60 amino acids. 3. There is another embryonic beta-like chain present in one haemoglobin component. It was designated omega and, in its first 54 amino acids, it has features that are more like avian globins than mammalian globins. 4. The one alpha-like embryonic globin sequenced has mammalian rather than avian characteristics. 5. A provisional phylogenetic tree of beta like globins has been determined. The Tammar epsilon-globin forms a monophyletic group with marsupial and other mammalian embryonic globins; the omega-globin forms a monophyletic group with bird adult and embryonic globins. PMID- 9750968 TI - Evolution of the human brain: is bigger better? AB - 1. The hominid brain has increased approximately three times in size since the Pliocene, but so has the brain of equids. The tripling of hominid brain size has been considered as an indicator of increased mental abilities, as it coincided with the production of tools, weapons and other artefacts of increasing sophistication. No indicators of the increase in equid intelligence are known. Intraspecific correlation between brain size and variously measured 'intelligence' is, in modern humans, very weak if not completely absent. With the exception of size, there are no major differences between the anatomy of ape and human brains. 2. A study of 297 estimates of body height, 626 estimates of bodyweight and 276 estimates of the cranial capacity of hominids dated at various periods over the past 5 million years shows that the increase in hominid brain size was paralleled by an increase in body size. 3. In a sample of 45 variously dated fossil hominids, brain size correlates isometrically with body size. 4. Since the Late Pleistocene (approximately 30,000 years ago), human brain size decreased by approximately 10%; yet again, this decrease was paralleled by a decrease in body size. 5. Therefore, it may be concluded that the gross anatomy of the hominid brain is not related to its functional capabilities. The large human brain:body size ratio may be a result of the structural reduction of the size of the gastrointestinal tract and, consequently, its musculoskeletal supports. It is related to richer, meat-based diets and extra-oral food processing rather than the exceptional increase in the size of the cerebrum. The exceptional mental abilities of humans may be a result of functional rather than anatomical evolution. PMID- 9750969 TI - Parathyroid hormone-related protein in lower vertebrates. AB - 1. Parathyroid hormone-related protein (PTHrP) is an important mediator of humoral hypercalcaemia of malignancy in humans. Normal human subjects have very low levels of PTHrP in their circulation. 2. Parathyroid hormone-related protein has recently been demonstrated in high levels in the circulation and tissues of the sea bream and the dogfish, leading to the hypothesis that PTHrP may be a 'classical' hormone in fish. 3. Immunohistochemistry and in situ hybridization were performed to investigate the evolutionary history of PTHrP. Tissues were examined from a number of lower vertebrates, including lungfish, lamprey and several species of bony and cartilaginous fish. Parathyroid hormone-related protein was localized to the skin and to kidney tubules in all animals studied. In the developing lungfish, PTHrP was observed in the notochord, developing brain and skeletal muscle layers. These results suggest that PTHrP is of ancient origin and has a basic and fundamental function in vertebrates. PMID- 9750970 TI - Studies of evolutionary temperature adaptation: muscle function and locomotor performance in Antarctic fish. AB - 1. Studies of evolutionary temperature adaptation of muscle and locomotor performance in fish are reviewed with a focus on the Antarctic fauna living at subzero temperatures. 2. Only limited data are available to compare the sustained and burst swimming kinematics and performance of Antarctic, temperate and tropical species. Available data indicate that low temperatures limit maximum swimming performance and this is especially evident in fish larvae. 3. In a recent study, muscle performance in the Antarctic rock cod Notothenia coriiceps at 0 degree C was found to be sufficient to produce maximum velocities during burst swimming that were similar to those seen in the sculpin Myoxocephalus scorpius at 10 degrees C, indicating temperature compensation of muscle and locomotor performance in the Antarctic fish. However, at 15 degrees C, sculpin produce maximum swimming velocities greater than N. coriiceps at 0 degree C. 4. It is recommended that strict hypothesis-driven investigations using ecologically relevant measures of performance are undertaken to study temperature adaptation in Antarctic fish. Recent detailed phylogenetic analyses of the Antarctic fish fauna and their temperate relatives will allow a stronger experimental approach by helping to separate what is due to adaptation to the cold and what is due to phylogeny alone. PMID- 9750971 TI - Aortic valve stenosis: a common clinical entity. PMID- 9750972 TI - Behavioral, cardiovascular, and neuroendocrine profiles following CCK-4 challenge in healthy volunteers: a comparison of panickers and nonpanickers. AB - Healthy subjects who panic following systemic cholecystokinin-tetrapeptide (CCK 4) challenge typically exhibit a symptom profile reminiscent of that evident among panic patients. However, the biological concomitants of CCK-4-induced panic in healthy subjects remain obscure. Accordingly, we evaluated the behavioral, cardiovascular, and neuroendocrine effects of CCK-4 in panickers and nonpanickers. Predictably, subjects who panicked with CCK-4 experienced more intense symptoms of panic and greater increases in ratings of fearful and anxious mood than did subjects who did not panic. CCK-4-induced increases in diastolic blood pressure, adrenocorticotropic hormone, prolactin, and growth hormone secretion were also significantly enhanced in subjects who panicked. The results of this study demonstrate that the behavioral experience of CCK-4-induced panic in healthy individuals is accompanied by marked biological changes and provide confirmation that CCK-4 is a useful model of panic for research among nonclinical subjects. PMID- 9750973 TI - Characteristics of agoraphobia in women and men with panic disorder with agoraphobia. AB - We compared female and male patients with panic disorder with agoraphobia (PDA) in terms of characteristics of agoraphobia (AG). Ninety-five patients (73 women and 22 men) with the SCID-based diagnosis of PDA were administered the National Institute of Mental Health Panic Questionnaire (NIMH PQ), and women and men were compared on the items of the NIMH PQ that pertain to AG and symptoms of panic attacks. Male and female patients did not differ significantly with respect to demographic characteristics, age of onset of panic disorder and AG, duration of PDA, and severity and frequency of symptoms experienced during panic attacks. Women avoided more situations than did men, but this difference was not statistically significant. Women avoided buses and being in unfamiliar places alone significantly more often. The only situation that was avoided more often by men, although not significantly, was staying at home alone. Women were significantly more likely to stay at home to avoid agoraphobic situations and significantly less likely to go outside of home alone. When going outside, women required a companion significantly more often. There were significantly more married women than married men who required a spouse as a companion, and significantly more women with children than men with children who required a child as a companion. Women thought that AG had affected the overall quality of their lives significantly more adversely. Whereas the overall "profile" of agoraphobic situations does not seem to distinguish between female and male patients with AG, females may be more impaired and appear more dependent than men in terms of requiring companions to move outside of the home. Cultural and psychological factors may be most likely to account for these findings. PMID- 9750974 TI - Effects of panic disorder treatments on personality disorder characteristics. AB - Ninety-three patients with panic disorder and mild or no agoraphobia were treated for their panic disorder by using either 11 sessions of individual cognitive behavior therapy or imipramine. Before and after treatment, their panic disorder symptomatology was assessed, and a self-report measure was administered to measure personality disorder characteristics [Klein et al., 1990: Wisconsin Personality Disorders Inventory]. In addition, some patients received this personality assessment again after six monthly maintenance sessions. Both treatments were equally effective in reducing panic disorder symptomatology, and both treatments had a positive influence on personality disorder characteristics. Personality disorder characteristics did not predict treatment outcome in either group. The implications of the findings for the assessment of personality and the treatment of panic disorder are discussed. PMID- 9750975 TI - Fluoxetine treatment of acral lick dermatitis in dogs: a placebo-controlled randomized double blind trial. AB - The aim of the study was to assess the efficacy and tolerability of fluoxetine treatment of acral lick dermatitis (ALD) in dogs and to investigate ALD as an animal model of obsessive-compulsive disorder (OCD). Sixty-three dogs with ALD were treated with fluoxetine 20 mg daily, or placebo, for 6 weeks. In the fluoxetine group, owners rated both appearance of the lesion (t = 10.2, df = 29, P < 0.0001) and licking behavior (t = 10.2, df = 29, P < 0.0001) as significantly improved by the end of the trial. Veterinarian-rated pre- and post-treatment photographs showed statistically significant improvement in the fluoxetine group (mean = 2.55). There were no significant changes in the placebo group as rated by owners and veterinarians. These results demonstrate the efficacy of fluoxetine in the treatment of ALD and lend further support to ALD as an animal model of OCD. PMID- 9750976 TI - Effect of a novel environment on resting heart rate in panic disorder. AB - Several studies have found higher resting heart rate among patients with panic disorder compared to healthy controls, whereas others have found no differences. It has been suggested that these differences may result from anticipatory anxiety. The purpose of this study was to compare the resting heart rates of 10 patients with panic disorder, 11 patients with social phobia, and 13 healthy controls during two consecutive visits to our laboratory. There were no significant differences between groups on resting heart rate on either day. However, patients with panic disorder did have significantly higher resting heart rates on day 1 versus day 2. This suggests that patients with panic disorder may experience greater anticipatory anxiety which is manifested in a higher resting heart rate than patients with social phobia or healthy controls. Implications for previous and future reports on resting heart rate measures in patients with panic disorder are discussed. PMID- 9750977 TI - Evidence from three fearful samples for a poor insight type in specific phobia. PMID- 9750978 TI - Associations with subsyndromal panic and the validity of DSM-IV criteria. AB - The purpose of this study was to compare subsyndromal panic--infrequent panic (IP) and limited symptom attacks (LSA)--with panic disorder (PD) in psychiatric comorbidity, quality of life (QOL), and health care utilization and to assess validity of DSM-III-R criteria for panic disorder. Randomly selected adults were screened for the presence of PD, IP, and LSA by using the Structured Clinical Interview of the DSM-IIIR. Subjects with panic symptoms and matched controls completed a structured interview concerning comorbidity, QOL, and utilization. Although PD and IP subjects reported more psychiatric comorbidity than did LSA subjects, LSA subjects had more comorbid conditions than did controls. Differences in utilization were limited to PD subjects. Although subsyndromal panic was associated with poor QOL, panic-related work disability was primarily seen in PD subjects. Regression analyses demonstrated little difference between LSA and IP subjects, but interaction analysis supported the distinction between LSA and full-blown panic attacks. Compared with controls, LSA and IP subjects had more psychiatric comorbidity. PD subjects also had poorer QOL and more utilization. Interaction analysis supports DSM-IV criteria for panic disorder. PMID- 9750979 TI - Use of the selective serotonin reuptake inhibitor citalopram in a possible animal analogue of obsessive-compulsive disorder. AB - Canine acral lick dermatitis (ALD) has been suggested as an animal analogue of obsessive-compulsive disorder (OCD). A series of dogs with ALD or similar conditions were treated with citalopram, the most selective of the selective serotonin reuptake inhibitors. Six of nine (66.7%) dogs showed significant improvement. Given the apparent efficacy of citalopram in the treatment of OCD and related disorders, these data provide further evidence that ALD is a useful animal analogue of OCD. PMID- 9750980 TI - Reactivation of posttraumatic stress disorder after minor head injury. AB - This report describes the reactivation of a posttraumatic stress disorder (PTSD) after a minor head injury in two young women who had recovered from extreme stress caused by sexual abuse during adolescence. Intrusive thoughts, images, dreams, and phobic avoidance bear a direct relationship to the specific circumstances of both head injury and sexual abuse, and were associated with obsessive-compulsive symptoms, generalized anxiety with panic, and depression. These findings suggest that in some individuals minor head injuries may induce not only extreme stress reactions, but also cause the reactivation of symptoms related to previous traumatic experiences. PMID- 9750982 TI - Cultural differences among health professionals: a case illustration. AB - This study illustrates that cultural differences arise among similarly trained health professionals. Health professionals must learn to communicate sensitively with colleagues from other cultures, to respect their values, and to recognize and resolve cultural differences that affect patient care. In this shrinking, multicultural world, health professionals cannot afford the comfortable illusion that all similarly trained practitioners share the same values about the care of patients and professional conduct. PMID- 9750981 TI - Deconstructing equity, autonomy, and ethical analysis. PMID- 9750983 TI - Barriers to completion of healthcare proxy forms: a qualitative analysis of ethnic differences. PMID- 9750984 TI - Cultural discrimination in mechanisms for health decisions: a view from New York. PMID- 9750985 TI - The family in medical decision making: Japanese perspectives. PMID- 9750986 TI - Heart transplantation selection criteria: attitudes of ethnically diverse medical students. PMID- 9750987 TI - The ethics of placebo-controlled trials for perinatal transmission of HIV in developing countries. PMID- 9750988 TI - The "best proven therapeutic method" standard in clinical trials in technologically developing countries. PMID- 9750989 TI - Cultural diversity and informed consent. PMID- 9750990 TI - Respect for autonomy and a couple's decision. PMID- 9750991 TI - A study of healthcare professionals' perspectives about a cross-cultural ethical conflict involving a Hmong patient and her family. PMID- 9750992 TI - Commentary: "missing" patients by seeing only their cultures. PMID- 9750993 TI - Futility and the goals of medicine. PMID- 9750994 TI - Resisting the siren: commentary. PMID- 9750995 TI - Legal trends in bioethics. PMID- 9750996 TI - Self-efficacy and substance abuse: assessment using a brief phone interview. AB - Considerable research has shown that one's self-efficacy to avoid cigarette smoking and alcohol use increases during treatment and that high self-efficacy ratings at follow-up are associated with positive outcome. The present study extends existing research in two ways. First, self-efficacy was assessed among a predominantly crack-cocaine-using population during treatment and 1 month following treatment. Second, the viability of a brief self-efficacy measure (4 item) was assessed using a phone interview. Results from 186 patients (61% reporting crack-cocaine) interviewed following treatment showed that self efficacy increased during treatment and was higher for patients reporting abstinence I month after treatment. The results from the brief self-efficacy assessment were comparable to an established version of the self-efficacy measure. These findings suggest that (a) self-efficacy may be related to the maintenance of abstinence from cocaine and other substances of abuse, and (b) self-efficacy can be measured quickly and reliably through a phone interview. PMID- 9750997 TI - Addressing trauma in substance abuse treatment with American Indian adolescents. AB - Substance abuse among American Indian adolescents is a serious problem that frequently continues into adulthood. Therefore, it is important to investigate all potential means of prevention and treatment of substance abuse that might improve the physical and psychological health it undermines. This paper examines the prevalence of substance abuse and its potential relationships with physical/emotional trauma or loss that occurs in American Indian adolescents' lives. The possible benefits of addressing trauma and posttraumatic stress as means of enhancing treatment is explored. An example of residential treatment that involved a focus on trauma and loss is included. PMID- 9750999 TI - Monitoring patterns of substance use in drug-dependent patients. AB - Drug-addicted patients (N = 435) admitted for treatment in different clinical settings were studied. Patients were classified according to their self-report of consumed drugs and to the results of urine screening tests. Of the patients, 77.8% were active consumers, 9.6% were included in a methadone maintenance program, and 12.6% were abstinent. In the active consumer patients, positive urine screening results surpassed by far the information provided in the self reports. Most patients tested positive to several drugs, while only 8.7% tested negative to all screened drugs. These results indicate that the information provided by drug-dependent patients lacks reliability when an analytical screening method is used simultaneously. PMID- 9750998 TI - Dynamic Recovery: comparative study of therapeutic communities in homeless shelters for men. AB - The Dynamic Recovery Project examined relationships between homelessness, substance abuse, and recovery, and investigated the effectiveness of the therapeutic community (TC) treatment model in helping homeless drug users move toward stable, drug-free living. This project compared two short-term TCs that were situated within pre-existing homeless shelters with a clean and sober dormitory. In a separate condition, peer counselors and staff were provided additional training in TC philosophy and practice to reduce program dropout. Dramatic decreases in drug and alcohol use at follow-up were verified by urinalysis. Length of time in treatment rather than specific program accounted for decreased alcohol and drug use. Important decreases in posttreatment criminality for both treatment programs were documented. The comparison group, starting with low criminality, experienced smaller, nonsignificant decreases unrelated to type of program or time in treatment. Major declines in Beck Depression Scores were evident, but were unrelated to groups or time in treatment. Training had no measurable impact on client retention or outcomes and there were no significant differences between TCs and the comparison group on posttreatment drug use, criminality, or depression. This report documents that short-term therapeutic communities can be successfully implemented in public shelters for homeless men. PMID- 9751000 TI - Laboratory exposure to cocaine cues does not increase cocaine use by outpatient subjects. AB - Sixty-nine cocaine-dependent outpatients were exposed to cocaine-related stimuli and to non-drug events on separate days. Cocaine cue sessions were always followed by a meeting with a trained clinician designed to eliminate any craving that remained following cue presentations. Urine samples were collected before each laboratory session and 1 to 3 days later. Neither rates of cocaine use nor average urine metabolite values differed following the two sessions. Nearly 90% of subjects had the same urine test result both before and after the cocaine cue session. Thus, laboratory presentation of cocaine cues to outpatient subjects did not increase their risk of subsequent drug-taking. These results suggest that with proper clinical protections, cue exposure can be used as a treatment outcome measure and a behavioral intervention in outpatient settings without increasing the risk of drug use. PMID- 9751001 TI - An evaluation of intravenous ethanol in hospitalized patients. AB - Alcohol withdrawal is a serious complication of heavy alcohol use and a condition requiring patient stabilization before initiating surgery or implementing lifesaving procedures for injury. Intravenous ethanol (IVE) is used to prevent withdrawal during these maneuvers. This report explores the use and potential problems of this practice in an academic urban medical center. This study was undertaken to improve the treatment of IVE recipients in an urban, academic health system providing trauma, surgery, and general inpatient services. All 68 patients, identified by a review of the pharmacy database for the period August 1993 through January 1994, received IVE during their stay. A priori outcome measures related to the course of therapy in the selected cases. Of all patients studied, 67.6% were admitted for alcohol-related trauma; 61.8% of IVE recipients had no documented risk factors for delirium tremens (59.5% of these were oriented); 17.6% were discharged on the same day the drip was discontinued; only 17.6% were referred to the alcohol consult team; and, throughout the course of therapy in all cases, no blood alcohol level (BAL) determinations were recorded in patients' records. The use of IVE is associated with potentially serious clinical concerns. We found a high prevalence of alcohol-related admissions, inconsistent IVE administration, and a low rate of alcohol consult requests. Guidelines to improve the selection, management, and disposition of IVE recipients are suggested. PMID- 9751002 TI - Stercoral perforation of the colon. A complication of methadone maintenance. PMID- 9751003 TI - Substance use disorder-PTSD comorbidity. Patients' perceptions of symptom interplay and treatment issues. AB - Forty-two patients with both a current substance use disorder (SUD) and posttraumatic stress disorder (PTSD) were asked about the interrelationship of their two disorders, their treatment preferences and experiences, as well as possible deterrents to receiving PTSD treatment. Patients perceived their two disorders to be functionally related. They reported that when one disorder worsened, their other disorder was more likely to worsen; when one disorder improved, the other disorder similarly improved. Consistent with these perceptions, SUD-PTSD patients favored simultaneous treatment of their two disorders. The majority of SUD-PTSD patients were never referred to PTSD treatment. Although several possible deterrents to PTSD treatment were identified, only lack of trust appeared to differentiate PTSD treatment compliers versus noncompliers. Implications of these findings on referral and treatment practices are discussed. PMID- 9751004 TI - Drug using offenders in south London. Trends and outcomes. AB - In 1993, the Home Office Drugs Prevention Initiative commissioned a study in Southwark, London of 112 drug misusing offenders referred by probation for treatment. There was a high incidence of relapse. After specialist intervention, 63% were identified as having reoffended, although the proportion of burglaries and violence was reduced. Discriminant Function analysis was used to assess whether contact with the probation service and a drug advice centre were significant determinants of preventing reoffending. Better rates of attendance with such agencies did reduce reconviction rates. Over half of the sample had used cocaine or crack. The comparatively high incidence of violent offences (17%) was related to the misuse of nonopiates. Conversely, sole use of opiates was a reliable predictor of nonviolence. Abstinence was only achieved by a small minority (5%). Over half of those assessed for intervention in the sample had recently appeared at Crown Court (higher level courts) with 75% having been sentenced to custody previously, this showed that the cohort was of relatively serious offenders. PMID- 9751005 TI - Reducing substance abuse during pregnancy. Discriminating among levels of response in a prenatal setting. AB - Providers in prenatal care settings are well-positioned to help pregnant women with substance abuse problems take the first steps toward recovery. This study reports the results of the ANGELS Program, a program of enhanced prenatal care designed to reduce substance use among pregnant women. In a suburban office serving a broad range of pregnant women, certified nurse-midwives (CNMs) and on site addictions counselors addressed substance abuse during prenatal care. This paper describes a cohort of 77 pregnant women who were identified as abusers of alcohol and/or other drugs at the start of pregnancy. According to a level of change rating assigned by the CNM at delivery, 51% of women were able to be largely abstinent during their pregnancy, 35% had reduced their use somewhat, and 14% had shown no change in use. Discriminant analysis techniques were used to learn what characteristics differentiated women in these three level of change groups. Baseline variables that differentiated the groups included severity of cocaine and cannabis use, psychosocial stressors, and initiation of prenatal care. Significant process variables included number of prenatal visits and contact with the addictions counselors. Clinical vignettes illustrate the differences among women in the three level of change groups. Implications of the results are discussed. PMID- 9751006 TI - Effects of a therapeutic camping program on addiction recovery. The Algonquin Haymarket Relapse Prevention Program. AB - A group of 13 men and women in substance abuse treatment participated in a 3-day residential program experience based on integrated principles from adventure therapy, therapeutic camping, and relapse prevention. The experimental group is compared to a group of 18 men and women who received the usual and customary relapse prevention program. Both groups completed pre- and postintervention questionnaires. There were no differences in drinking-related locus of control, stress, or problem-solving between groups at postinterview, but there were significant improvements in autonomic arousal, frequency of negative thoughts, and alcohol craving. Participants in both groups were interviewed 10 months after the 3-day intervention. Considering individuals who were unreachable as relapsed, the 10-month follow-up relapse rate was 31% for the experimental group and 58% for the comparison group. These results add to the limited body of research supporting outdoor adventure and therapeutic camping experiences integrated with traditional relapse prevention activities as an adjunct to substance abuse treatment. PMID- 9751007 TI - Chronic hepatitis C: the virus, its discovery and the natural history of the disease. AB - The identification of hepatitis A and hepatitis B led to the recognition that a third virus was capable of causing blood-borne hepatitis. The pathogen responsible for this nonA, nonB hepatitis was identified in the late 1980s and subsequently named hepatitis C. Since the discovery of hepatitis C there has been a pandemic of research publications describing the natural history of the infection and it is now known that this virus can cause serious liver damage in a proportion of infected patients. It is now clear that the effects of infection with hepatitis C and alcohol misuse are additive and that there is an increased risk of hepatic complications in infected patients who abuse alcohol. PMID- 9751008 TI - Report from the International Symposium on viral hepatitis, 24-25 October 1997, Warsaw, Poland. AB - The authors present the results of an International Symposium on 'Viral Hepatitis', held in Warsaw on 24-25 October 1997 and dedicated to the scientific activity of Professor Adam Nowoslawski, the founder of the Polish school of Immunopathology, with many contributions to the viral hepatitis research. The symposium was divided into main sessions and poster reports which covered most of the topics in this field. This successful meeting has gathered many distinguished speakers from different countries and was attended by ca. 350 participants, mainly from Poland, but also from the neighbouring countries. PMID- 9751009 TI - Degree and distribution of variability in the 5' untranslated, E1, E2/NS1 and NS5 regions of the hepatitis C virus (HCV). AB - Hepatitis C virus (HCV) shows a high degree of variability resulting in many different variants. In this work we described the variability of several subgenomic fragments from the 5' untranslated region (5'-UTR) and E1, E2/NS1 and NS5 regions comparing, for every position, all the sequences published in GenBank v. 88 (July 1995) as well as new sequences obtained in this work. Variability was determined in two ways. First, we analysed the degree and type of substitutions found in these regions. Second, we defined the most variable and conserved segments in each region and compared our prediction with previous studies. Our results confirm that HCV variability changes along the different regions. Although we found four variable domains in the 5'-UTR, this region was the only one to contain conserved domains. Envelope (E1, E2/NS1) and NS5 regions showed high variability throughout; however, we were able to define six and three hypervariable domains, respectively. The degree and distribution of variability established in this work is supported by the high number of sequences and the different types included in the study. Knowledge of how variability is distributed along the different regions of the HCV genome could be of use in the design of new diagnostic and therapeutic strategies against HCV infection. PMID- 9751010 TI - Incidence and clinical significance of hepatitis B virus precore gene translation initiation mutations in e antigen-negative patients. AB - Hepatitis Be antigen (HBeAg)-negative chronic hepatitis B (CHB) is associated with hepatitis B virus (HBV) variants harbouring changes in the precore region. Most commonly, a G to A point mutation at nucleotide 1896 (m1896) creates a novel translation stop codon that prevents HBeAg production. In the Mediterranean region the m1896 mutation prevails in greater than 98% of HBeAg-negative CHB patients. In this study the prevalence of additional mutations in the precore region was investigated among patients with chronic HBV infection. Precore sequences were determined by sequencing serum HBV DNA amplified by polymerase chain reaction (PCR) with primers flanking the precore/core region. Thirty-one HBeAg-negative and five HBeAg-positive individuals were studied. All HBeAg negative patients (100%) harboured the m1896 mutation and 20 (64.5%) also had a G to A mutation at nucleotide 1899 (m1899). Additional mutations affecting the translation initiation of the precore gene were found in seven (22.5%) patients, all with active liver disease, five of whom had episodes of HBV reactivation. HBeAg-positive patients had no mutations in these positions and neither did any of the five BHeAg-negative patients with normal levels of liver enzymes, representing the healthy carrier state of HBV infection. Serial sample analysis from one patient revealed that the initiation codon mutation developed following HBeAg seroconversion and the appearance of m1896. During periods of high HBV replication, the ratio of mutant to wild-type ATG was found to increase in parallel with HBV DNA levels. These data show that a significant proportion of HBeAg-negative patients who already harbour the 1896 stop codon mutation may subsequently develop precore translation initiation mutations, which appear to be associated with enhanced HBV replication and severe liver disease. PMID- 9751011 TI - Response to interferon therapy: influence of human leucocyte antigen alleles in patients with chronic hepatitis C. AB - The response to interferon (IFN) therapy in patients with chronic hepatitis C is characterized by normalization of the serum alanine aminotransferase (ALT) activity during treatment, but relapse within 6 months of cessation of therapy is common. Viral characteristics, such as the genotype and viral load, may influence the patient response to IFN. The aim of this study was to examine host factors, namely the genetically determined human leucocyte antigen (HLA) class II alleles, in patients with chronic hepatitis C, and their relationship to the response to IFN therapy. Seventy white patients with chronic hepatitis C, treated with IFN alpha for 6 months, were enrolled in the study. Serum ALT was measured at the end of treatment to assess short-term response and again 6 months post-treatment to assess sustained response. Sequence-specific primers were used in the polymerase chain reaction (PCR) to amplify genomic DNA isolated from peripheral mononuclear cells. HLA class II alleles were determined by analysis of the amplicon by gel electrophoresis and hybridization of sequence-specific oligonucleotide probes. At the end of treatment, 25 of the 70 patients (36%) had a normal ALT. By 6 months post-treatment, only six patients (9%) had a sustained normalization of ALT. The frequency of the allele DRB1*0404 was significantly higher in patients with a sustained response as compared to those lacking such a response (25.0% vs 2.3%, with a Bonferonni-corrected P-value of 0.019). There was no difference in the frequency of other class II alleles at the DRB1 and DQB1 loci in responders as compared with non-responders. Therefore, we conclude that the maintenance of a response to IFN in chronic hepatitis C may be, in part, determined by genetic factors in the host. PMID- 9751012 TI - Prediction of cirrhosis in patients with chronic hepatitis C infection by artificial neural network analysis of virus and clinical factors. AB - The diagnosis of cirrhosis in patients with hepatitis C virus (HCV) infection is currently made using a liver biopsy. In this study we have trained and validated artificial neural networks (ANN) with routine clinical host and viral parameters to predict the presence or absence of cirrhosis in patients with chronic HCV infection and assessed and interpreted the role of the different inputs on the ANN classification. Fifteen routine clinical and virological factors were collated from 112 patients who were HCV RNA positive by reverse transcriptase polymerase chain reaction (RT-PCR). Standard and Ward-type feed-forward fully connected ANN analyses were carried out both by training the networks with data from 82 patients and subsequently testing with data from 30 patients plus performing leave-one-out tests for the whole patient data set. The ANN results were also compared with those from multiple logistic regression. The performance of both ANN methods was superior compared with the logistic regression. The best performance was obtained with the Ward-type ANNs resulting in a sensitivity of 92% and a specificity of 98.9% together with a predictive value of a positive test of 95% and a predictive value of a negative test of 97% in the leave-one-out test. Hence, further validation of the ANN analysis is likely to provide a non invasive test for diagnosing cirrhosis in HCV-infected patients. PMID- 9751013 TI - A comparative trial of two surface subunit recombinant hepatitis B vaccines vs a surface and PreS subunit vaccine for immunization of healthy adults. AB - Hepatitis B virus (HBV) infection is the leading cause of chronic hepatitis and cirrhosis in Turkey. The prevalence of hepatitis B surface antigen (HBsAg) positivity in Turkey is 5 to 10%. HBV is almost completely preventable with the use of hepatitis B vaccines. The most commonly used vaccine is that which contains the predominant viral surface (S) polypeptide. It elicits protective antibodies in greater than 90% of healthy subjects. A vaccine containing the PreS1 and PreS2 antigenic domains has recently been reported as being more efficient in achieving successful immunization in individuals who have not previously responded to the isolated S-antigen vaccine. In this study, the efficacy of a S and PreS-containing vaccine was compared with that of two different standard isolated S-antigen-containing vaccines in terms of the immunization protection produced against HBV in normal healthy adults who had not previously been immunized. Seventy-six young adults (aged 17-22) were randomly assigned to receive 1 ml (20 micrograms) of either one of two standard S-subunit recombinant hepatitis B vaccines (Engerix B. or Hepavax) or the combined S and PreS subunit vaccine (Gen Hevac B) intramuscularly in the deltoid muscle at 0, 1 and 2 months. Hepatitis B surface antigen antibody titres were measured at 1, 2 and 12 months. A titre > or = 10 IU ml-1 was considered to be protective. All subjects receiving the two standard isolated S-antigen-containing vaccines responded to the vaccination with reasonable antibody titres. One-half to two thirds of those vaccinated developed high antibody titres (> 100 IU ml-1). In contrast, 9% of those receiving the combined PreS1 and PreS2 plus S antigens failed to respond, as demonstrated by antibody titres below the level considered to be protective. The mean titres at 12 months were 107 +/- 12 IU ml-1 (Engerix B), 102 +/- 12 IU ml-1 (Gen Hevac B) and 117 +/- 12 IU ml-1 (Hepavax Gene). Hence, no important difference in term of response to vaccination was found between the two different types of vaccines. As recombinant S-subunit vaccines are less expensive than those that combine S and PreS antigens, it is suggested that, when immunizing normal healthy adults, a standard isolated S-antigen containing vaccine should be used. PMID- 9751014 TI - Relationship between biochemical and virological responses to interferon therapy in chronic hepatitis C infection. Consensus Interferon Study Group. AB - We have investigated the relationship between serum alanine aminotransferase (ALT) and hepatitis C virus (HCV) RNA in the assessment of responses to interferon (IFN) therapy in chronic HCV infection. Data from 704 patients with HCV infection who were randomized to receive consensus IFN-alpha (CIFN) 3 micrograms (n = 232 patients) or 9 micrograms (n = 232 patients), or IFN-alpha 2b 3 million units (MU) (n = 240 patients), were used for these analyses. All patients were treated three times weekly. Hepatitis C viral RNA (HCV RNA) was determined by quantitative reverse transcriptase-polymerase chain reaction (RT PCR) with a lower limit of detection of 100 copies ml-1. Of patients with normal serum ALT concentrations, 53% (120/225) had undetectable HCV RNA at the end-of treatment period and 47% (51/109) had undetectable HCV RNA at the end of the post treatment observation period. In contrast, of the patients with undetectable HCV RNA, 75% (120/161) and 84% (51/61) had normal serum ALT activities at the end-of treatment and post-treatment observations periods, respectively. The majority of patients with undetectable HCV RNA had normal ALT values. In contrast, only half of the patients with normal ALT values were negative for HCV. End-of-treatment HCV RNA response also better predicted sustained virological response than did end-of-treatment ALT response. PMID- 9751015 TI - Prevalence and determinants of hepatitis A virus exposure among prison entrants in Queensland, Australia: implications for public health control. AB - In September 1997, a multicentre outbreak of hepatitis A virus (HAV) infection occurred in the Queensland prison system following a prolonged community-based HAV epidemic among illicit drug users. As part of the public health response, a cross-sectional survey was undertaken to estimate the sero-prevalence of, and identify the determinants for, recent and past HAV infection among the incoming male prisoner population. Exposure data were collected through face-to-face interviews with 214 consenting inmates, whose sera were screened for immunoglobulin G (IgG) and immunoglobulin M (IgM) antibodies to HAV. Overall, 81 (37.9%) inmates were HAV-IgG seropositive, and four inmates were HAV-IgM seropositive, HAV-IgG seropositivity was strongly associated with year of birth (age) (Ptrend < 0.0001), being born outside Australia (relative risk (RR) 1.9, 95% CI 1.4-2.7) and being of a non-English speaking background (RR 2.5, 95% CI 1.7-3.7). Lifestyle exposures (such as occupation, overseas travel and illict drug use) were not associated with an increased risk of HAV-IgG seropositivity. In contrast, all four HAV-IgM seropositive inmates were English-speaking, Australian-born males who used illicit drugs. These findings suggest that the risk factors for recent and past HAV infections among prisoners differ, with implications for HAV control in correctional settings. Strategies for HAV prevention, including routine screening of inmates and vaccination of susceptibles, are considered in the context of current models of disease transmission. PMID- 9751016 TI - Nanoflow solvent gradient delivery from a microfabricated device for protein identifications by electrospray ionization mass spectrometry. AB - Microfabrication technology offers the opportunity to construct microfluidic modules which are designed to perform specific, dedicated functions. Here we report the construction of a microfabricated device for the generation and delivery by electroosmotic pumping of solvent gradients at nanoliter per minute flow rates. The device consists of three solvent reservoirs and channels which were etched in glass. Solvent gradients and solvent flows were generated by computer controlled differential electroosmotic pumping of aqueous and organic phase, respectively, from the solvent reservoirs. The device was integrated into an analytical system consisting of the solvent gradient delivery module, a reverse phase microcolumn and an electrospray ionization ion trap mass spectrometer (MS). The system was used for the analysis at high sensitivity of peptides and peptide mixtures generated by proteolytic digestion of proteins. We have measured an absolute limit of detection as low as 1 fmol and a concentration limit of detection at the 100 amol/microL level. The system was also successfully used for the identification of proteins separated by 1D and 2D gel electrophoresis. This was achieved by gradient frontal analysis of the peptide mixture generated by proteolysis of the respective proteins, and the automated generation and interpretation of collision-induced dissociation spectra. PMID- 9751017 TI - An integrated microfluidics-tandem mass spectrometry system for automated protein analysis. AB - We describe an integrated analytical system consisting of a microfluidics device micromachined using photolithography/etching technology, a panel of computer controlled high-voltage relays, and an electrospray ionization tandem mass spectrometer. Movement of solvents and samples on the device and off the device to the mass spectrometer was achieved by directed electroosmotic pumping induced by the activation of a suitable constellation of high-voltage relays. The system was used for the sequential automated analysis of protein digests. We demonstrate low femtomole per microliter sensitivity of detection and compatibility of the system with the automated analysis of proteins separated by two-dimensional gel electrophoresis. PMID- 9751018 TI - A microcapillary column switching HPLC-electrospray ionization MS system for the direct identification of peptides presented by major histocompatibility complex class I molecules. AB - A microcapillary column switching high-performance liquid chromatography (HPLC) system was developed for the separation of major histocompatibility complex (MHC) class I associated peptides. Combination of the column switching system with electrospray ionization mass spectrometry (ESIMS) enabled the detection and identification of the peptides at low-femtomole levels. Sample volumes of 30-50 microL were injected and concentrated onto a short, 100-micron-i.d. precolumn. The precolumn was coupled to a 100-micron-i.d. reversed-phase analytical HPLC column via a six-port valve. Peptides were separated on the analytical column using an ESI-compatible mobile phase at a flow rate of 0.5 microL/min. Peptides were eluted directly into the ESI source of either a magnetic sector MS or an ion trap MS. Peptides associated with human leukocyte antigen A*0201 molecules were determined in immunoaffinity-purified extracts from either measles virus infected cells or uninfected cells by microcapillary column switching HPLC-ESIMS. The approach toward detection of virus-specific peptides we used was based on the comparison of ion chromatograms obtained from the LC-MS analysis of extracts from virally infected cells and their uninfected counterparts. In this way, the molecular mass of peptides unique to virus infected cells was obtained. The utility of the system is demonstrated by the identification of a candidate epitope. Microcapillary column switching HPLC was used along with ESI ion trap tandem MS to identify the naturally processed viral peptide KLWESPQEI. This peptide was found to derive from the measles virus nonstructural protein C. The approach described here provides a versatile and sensitive method for the direct identification of viral peptides associated with MHC class I molecules. PMID- 9751019 TI - Reduction of fatty acid methyl esters to fatty alcohols to improve volatility for isotopic analysis without extraneous carbon. AB - Carbon in derivatization groups cannot be distinguished from analyte carbon by chromatography-based high-precision compound-specific or position-specific isotope analysis. We report the reduction of fatty acid methyl esters to fatty alcohols to facilitate high-quality chromatographic separation, without addition of extraneous carbon, with subsequent high-precision position-specific isotope analysis. Methyl palmitate is quantitatively reduced to 1-hexadecanol by LiAlH4 in a one-step reaction. Gas-phase pyrolysis of 1-hexadecanol results in a series of monounsaturated alcohols and alpha-olefins analogous to fragmentation found for methyl palmitate, as well as an additional peak corresponding to the pyrolytic dehydration product, 1-hexadecene. Carbon isotope analysis of the fragments yielded precision of SD (delta 13C) < 0.4/1000. Results of position specific analysis of very low enrichment [1-13C]-1-hexadecanol (delta 13C = 4.00/1000) showed no evidence of scrambling of the C1 position, and isotope ratios in accord with expectations. The pyrolysis product 1-hexadecene was isotopically enriched relative to 1-hexadecanol, which may cause minor depletion of other pyrolysis products that can be taken into account by routine calibration. The procedure is general and can be extended to compound-specific and position-specific analysis of moderate molecular weight, low-volatility analytes containing acid groups that would otherwise be blocked with methyl, ethyl, acetyl, or trimethyl silyl groups containing extraneous carbon. PMID- 9751020 TI - On-probe solid-phase extraction/MALDI-MS using ion-pairing interactions for the cleanup of peptides and proteins. AB - Samples originating from biological sources often contain a complex mixture of inorganic salts, buffers, chaotropic agents, surfactants/detergents, preservatives, and other solubilizing agents. However, the presence of these contaminants virtually ensures the failure of any subsequent analysis of the sample by matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS). Sample cleanup procedures, therefore, must be performed prior to MALDI-MS analysis. This paper reports a probe-surface derivatization method that greatly simplifies this sample preparation process. MALDI probes possessing self assembled monolayers (SAMs) terminated with ionic functional groups can rapidly extract peptides/proteins via ionic interactions from < or = 1-microL volumes of sample solutions placed directly on their surface. We have found that MALDI probes modified in this manner are a practical solution for analyzing very small volumes of peptide/protein solutions contaminated with high levels of inorganic salts, buffers, detergents, chaotropic agents, and other solubilizing agents. PMID- 9751021 TI - Isotachophoretic separations on a microchip. Normal Raman spectroscopy detection. AB - Isotachophoretic separations of the herbicides paraquat and diquat are performed in a glass microchip etched channel and monitored on-chip by normal Raman spectroscopy. The 40-micron-wide and 75-micron-deep separation channels are chemically etched in a serpentine design to 21-cm total length. A 120-micron thick glass cover slip is used to seal the channels. Separation field strengths up to 380 V/cm are used. The microchip is directly coupled to a Raman microprobe. No interfacing is required. Raman spectra are generated with a 2-W, 532-nm NdY VO4 laser and collected at 8-cm-1 resolution with a holographic transmissive spectrograph and a cryogenically cooled CCD. Data acquisition is at 2-5 spectra/s. Raman isotachopherograms of the pesticides at starting concentrations as low as 2.3 x 10(-7) M (60 ppb paraquat/80 ppb diquat) are presented. PMID- 9751022 TI - Fabrication of nanocolumns for liquid chromatography. AB - This paper shows that in situ micromachining can be used to simultaneously position and define (i) support particles, (ii) convective transport channels, (iii) an inlet distribution network of channels, and (iv) outlet channels in multiple chromatography columns on a single quartz wafer to the level of a few tenths of a micrometer. Stationary phases were bonded to 5 x 5 x 10 microns collocated monolith support structures separated by rectangular channels 1.5 microns wide and 10 microns deep with a low degree of deviation of channel width between the top and bottom of channels. High aspect ratio microfabrication can only be achieved with deep reactive ion etching. The volume of a 150 microns x 4.5 cm column was 18 nL. Column efficiency was evaluated in the capillary electrochromatography (CEC) mode using rhodamine 123 and a hydrocarbon stationary phase. Plate heights in these columns were typically 0.6 micron in the nonretained and 1.3 microns in the retained modes of operation. Columns were designed to have identical mobile-phase velocity in all channels in an effort to minimize outgassing during operation. When the total lateral cross-sectional area of channels at all points along the separation axis is identical, linear velocity of the mobile phase in a CEC column should be the same. Columns were operated at atmospheric pressure. PMID- 9751023 TI - Analysis of recombinant human platelet-derived growth factor by reversed-charge capillary zone electrophoresis. AB - Reversed-charge capillary zone electrophoresis (RC-CZE) has been developed as a clipping (proteolysis) assay for homodimeric protein recombinant human platelet derived growth factor (rhPDGF-BB), a major serum mitogenic factor involved in subcutaneous wound healing. When expressed in yeast, the protein is excreted as a fully folded homodimeric protein consisting of two antiparallel B chains held together by two interchain disulfide bonds. During fermentation, internal proteolysis (clipping between residues Arg32 and Thr33) and C-terminal truncation (Arg32 and Thr109) may occur. Internal proteolysis yields three potential forms of rhPDGF-BB: intact (both B chains are intact), single-clipped (one B chain is clipped), and double-clipped (both B chains are clipped). Clipping also creates new C-terminal sites for further C-terminal truncations and leads to a very complex mixture of isoforms. Routine baseline resolution of these three forms by various modes of HPLC proved unsuccessful. When the disulfide bonds of antiparallel chains are reduced, the complex peptide mixture can be analyzed by RP-HPLC; however, only the level of total clipping is identified. Since RC-CZE separation relies upon differences in molecular charge/size ratio, it can resolve the three rhPDGF-BB forms differing in the additional exposed residues. The choice of reversed-charge CZE columns (amine-coated column) allows proteins of high pI such as rhPDGF-BB (pI > 10) to be readily analyzed while minimizing protein loss from column adsorption. To simplify the electropherogram of clipped forms, the sample is treated first with carboxypeptidase B to reduce the charge microheterogeneity of partial Arg32 truncation. Analysis of rhPDGF-BB by RC-CZE yields a baseline separation between the three forms, intact and single- and double-clipped rhPDGF-BB. PMID- 9751024 TI - General protease assay method coupling solid-phase substrate extraction and capillary electrophoresis. AB - Capillary electrophoresis with laser-induced fluorescence detection was used to develop a universal, highly specific protease assay. In this method, a peptide, biotinylated at the N-terminus, is labeled with fluorescein at a lysine residue near the C-terminus. Impurities are removed from the fluorescence labeling mixture by solid-phase extraction of the substrate on immobilized streptavidin, followed by extensive washing. The purified fluorescent substrate is dissociated from the streptavidin and incubated with the protease. The peptide sequence between the biotin and fluorescent label contains the cleavage sequence of the protease of interest. After cleavage, the fluorescent product does not contain a biotin group. A second solid-phase extraction is used to remove unreacted substrate to dramatically lower the background signal. The product is detected by capillary electrophoresis, which provides powerful discrimination against products generated by nonspecific proteases. With chymotrypsin as a test protease, product was detected with as little as 10 pg/mL (4.6 x 10(-13) M) chymotrypsin, or 5 amol of enzyme in the 10-microL sample volume. PMID- 9751025 TI - Enantioselective gas chromatography/mass spectrometry of methylsulfonyl PCBs with application to arctic marine mammals. AB - Four different commercially available cyclodextrin (CD) capillary gas chromatography (GC) columns were tested for the enantioselective separation of nine environmentally persistent atropisomeric 3- and 4-methylsulfonyl PCBs (MeSO2 CBs). The selected columns contained cyclodextrins with various cavity diameters (beta- or gamma-CD), which were methylated and/or tert-butyldimethylsilylated (TBDMS) in the 2,3,6-O-positions. The beta-CD column with TBDMS substituents in all of the 2,3,6-O-positions was by far the most selective column for the MeSO2 CBs tested. Enantiomers of congeners with 3-MeSO2 substitution were more easily separated than those with 4-MeSO2 substitution. The separation also seemed to be enhanced for congeners with the chlorine atoms on the non-MeSO2-containing ring and clustered on one side of the same ring. The 2,3-di-O-methyl-6-O-TBDMS-beta-CD was found to give somewhat better selectivity than the corresponding gamma-CD, in comparison between the two columns, which were identical in all other respects. Enantioselective analysis of arctic ringed seal (Phoca hispida) and polar bear (Ursus maritimus) adipose tissue revealed a strong dominance of certain enantiomers. For example, the enantiomer ratio (ER) of 3-MeSO2-CB149 was 0.32 and < 0.1 in ringed seal blubber and polar bear fat, respectively. These low ER values are indicative of highly enantioselective formation, enantioselective metabolism, enantioselective transport across cell membranes, or a combination of the three in both species. Comparable results for the enantiomeric analysis of MeSO2-CBs in biotic tissue extracts were obtained using two highly selective mass spectrometric techniques, ion trap mass spectrometry/mass spectrometry and electron capture negative ion low-resolution mass spectrometry. PMID- 9751026 TI - Viral characterization by direct analysis of capsid proteins. AB - Matrix-assisted laser desorption/ionization mass spectrometry has enabled viral coat proteins to be characterized directly from the virus. This analysis, demonstrated here with tobacco mosaic virus U2, a bacteriophage MS2, and equine encephalitis TRD, is achieved with a combination of organic acid, UV-absorbing matrix, and high-energy desorption with a nitrogen laser. The molecular weights of these proteins are determined with sufficient accuracy to allow differentiation among viral species and strains. The abundant hydrophobic MS2 coat protein was analyzed in aliquots of culture medium and of the tobacco mosaic virus coat protein in infected leaves. This method provides rapid detection of coat protein in the low-femtomole range, as estimated by titering plaque-forming units of MS2. PMID- 9751027 TI - Miniaturized carbon monoxide sonde for atmospheric measurements. AB - The design and test results of a simple carbon monoxide detector based on the reducing gas detector principle are presented. This instrument has several features which distinguish it from similar carbon monoxide analyzers, all of which contribute to its usefulness on small airborne platforms such as kites, balloons, and light aircraft. Compact size, battery operation, low cost, and light weight are among the most important improvements made over to earlier systems. The instrument fits a package 10 x 20 x 25 cm, has a mass of 2.0 kg, and consumes an average of 20 W of power. This instrument will, therefore, offer a means of incorporating carbon monoxide measurements in recoverable or disposable balloon sondes. The instrument makes an independent measurement of carbon monoxide every 8 s, with a sensitivity of 3 ppbv carbon monoxide (limit of detection for S/N = 3) and a precision of 4%. PMID- 9751028 TI - Enhancement of molecular fluorescence near the surface of colloidal metal films. AB - Fluorescence enhancement was studied on silver colloidal metal films (CMFs) using two systems: (1) Langmuir--Blodgett monolayers of fluorescein-labeled phospholipids separated from the surface of the films by spacer layers of octadecanoic acid and (2) biotin--fluorescein conjugates captured by avidin molecules adsorbed on top of a multilayer structure formed by alternating layers of bovine serum albumin--biotin conjugate (BSA--biotin) and avidin. The dependence of fluorescence intensity on the number of lipid or protein spacer layers deposited on the surface of the CMF was investigated. The results demonstrate the requirement for adsorbate location within the region between Ag particles for maximal enhancement. The density of avidin molecules on the surface of the BSA--biotin/avidin multilayers adsorbed on the CMF was also determined. A procedure for forming a rigid, uniform silica layer around the Ag particles on the CMF is described. The layer protects the particles from undesirable chemical reactions such as etching by halide ions, for example, and provides the requisite stability for bioanalytical applications. Colloidal films composed of Ag particles covered by approximately 10-nm-thick silica layers were tested for fluorescence enhancement using goat immunoglobulin and a conjugate of rabbit anti goat immunoglobulin with 6-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino)hexanoate. An enhancement factor of approximately 20 was obtained. PMID- 9751029 TI - Fiber bundle based scanning detection system for automated DNA sequencing. AB - High-throughput DNA sequencing techniques are under rapid development currently, mainly triggered by the Human Genome Project. At the present time, slab gel based automated DNA sequencing is the standard procedure, utilizing fluorophore labeling and laser-induced fluorescence detection with scanning technology. In this paper, a novel, fiber-optic bundle based detection system is introduced, where a central illuminating fiber is used for the excitation of the electrophoretically separated fluorophore-labeled DNA sequencing fragments, along with several collecting fibers disposed around the illuminating fiber to collect the emitted fluorescent signal. As a model system, Cy5-labeled DNA sequencing fragments were separated on an ultrathin polyacrylamide slab gel and detected by the fiber bundle based laser-induced fluorescence detection system. A 640-nm diode laser was used to generate the illumination beam, and the emitted light collected by the fiber bundle was detected by a solid-state avalanche photodiode. PMID- 9751030 TI - Determination of halogenated hydrocarbons by helium microwave plasma torch time of-flight mass spectrometry coupled to gas chromatography. AB - A helium microwave plasma torch (MPT) was coupled to time-of-flight mass spectrometry (TOFMS) for the detection of halogenated hydrocarbons separated by capillary gas chromatography (GC). The GC-MPT-TOFMS system offered excellent stability over the course of the experiments and avoided mass spectral peak distortions caused by spectral skew. In the initial studies, empirical formulas based on the halogen-to-carbon ratio were predicted utilizing a flow cell apparatus. The MPT proved to be very robust and could handle large amounts of organic vapor. Results from this study indicate that, for both aromatic and aliphatic halogenated hydrocarbons, the ratios of carbon to chlorine signals correlate well (r = 0.994) with the ones expected from their chemical composition. This study was later extended to include chromatographic separation. For a series of homologous aliphatic halogenated hydrocarbons, a correlation coefficient of 0.999 was obtained for both peak heights and peak areas obtained from a single chromatogram. A novel Nichrome wire-heated transfer line was developed to ensure that the capillary column was heated efficiently from the GC oven to the MPT and then through the length of the MPT up to the microwave plasma itself. No appreciable peak broadening and no detectable memory effects were associated with the heated transfer line. The GC-MPT-TOFMS system offered equal sensitivity for I, Br, and Cl. Absolute detection limits for the halogenated hydrocarbons ranged from 160 to 330 fg, constituting an improvement by a factor of 5-35 over earlier results obtained with MIPs supported in a TM010 cavity and combined with quadrupole-based mass spectrometry. In addition, the effect of molecular gases on the MPT performance was investigated. Up to about 1% (v/v) of either oxygen or hydrogen in the central channel helium flow attenuated the signal levels for both carbon and chlorine, with the larger loss seen in the chlorine signal. PMID- 9751031 TI - A fluorescent temperature probe based on the association between the excited states of 4-(N,N-dimethylamino)benzonitrile and beta-cyclodextrin. AB - A temperature probe based on the fluorescence properties of the two excited states of 4-(N,N-dimethylamino)benzonitrile (DMABN) in equilibrium with beta cyclodextrin (CD) in aqueous solution is presented. The fluorescence intensity of the Franck-Condon excited state (FB) as a function of temperature shows a straight line with a correlation better than 0.99 in the 283-308 K temperature interval. On the other hand, the fluorescence intensity of the twisted internal charge-transfer state (FA) remains constant in the same temperature interval because the binding of DMABN in the A* state to CD is isoenthalpic and entropy driven. It is found that the FA/FB ratio is independent of the excitation intensity at a specified temperature, shows a linear relationship with temperature, and allows temperature measurements with a resolution of +/- 2.5 K. PMID- 9751032 TI - DDE mercury, and selenium in Biota, sediments, and water of the Rio Grande-Rio Bravo Basin, 1965-1995. AB - An assessment of contaminant stressors on biota of the Rio Grande was conducted to identify relevant contaminant issues, assess exposure and ecological effects, identify data gaps, and determine potential risks. Most contaminant data were from studies conducted during 1965-1995 in the Lower Rio Grande, on the Texas side of the river, within a 100-km boundary from Falcon Dam to the mouth. Contaminants most frequently reported were organochlorine compounds (OCs) and trace elements. The number of records for OCs and trace elements was at least twofold greater for fish than for birds, mammals, or reptiles. Of the OCs, p,p' DDE was the most commonly reported. Among the trace elements, Hg was one of the most frequently reported; however, Se, As, Pb, Cu, and Zn were also common. The highest concentrations of OCs and trace elements were reported predominantly from Lower Rio Grande Valley locations, with approximately 68% of the highest values detected from Falcon Dam to the mouth of the river. Twenty-six (20%) of the locations with maximum concentrations corresponded to portions of Llano Grande Lake and the Arroyo Colorado. Recent analyses of birds and fish indicate that levels of DDE are currently much lower than in the 1970s or 1980s in Rio Grande wildlife. This apparent decline does not apply to Hg and Se levels in birds and fish, which have remained more or less constant, but may have increased over the years in some locations. Hg was of particular concern because of high levels found recently in addled eggs of aplomado falcons and in their potential prey. Hg was elevated in fish from the Big Bend area. Also, Se in fish sampled in 1993 and 1994 was near or above the threshold for potential effects in fish-eating wildlife. Future investigations should evaluate the potential impacts of Hg and Se on aquatic and terrestrial species from selected sites of concern. PMID- 9751033 TI - Environmental chemistry and toxicology of polychlorinated n-alkanes. AB - Polychlorinated-n-alkanes (PCAs) or chlorinated paraffins consist of C10 to C30 n alkanes with chlorine content from 30% to 70% by mass. PCAs are used as high temperature lubricants, plasticizers, flame retardants, and additives in adhesives, paints, rubber, and sealants. This review presents the existing data on the environmental chemistry and toxicology of PCAs and a preliminary exposure and risk assessment. There is limited information on the levels, fate, or biological effects of PCAs in the environment. This results both from the difficulty associated with quantifying PCAs, because of the complexity inherent to commercial formulations, and from the limited knowledge of their physicochemical properties and biodegradation rates. There are indications that PCAs are widespread environmental contaminants at ng/L levels in surface waters and ng/g (wet wt) levels in biota. However, environmental measurements of PCAs are very limited in the U.S. and Canada, and are only slightly more detailed in western Europe. Assuming that reported water concentrations are mainly caused by the short chain (C10-C13) compounds, aquatic organisms may be at risk from exposure to PCAs. Fugacity level II modeling for two representative PCAs, using the best available physicochemical property data and estimated degradation rates, suggested that C16C24Cl10 would achieve higher concentrations in biota, sediment, and soil than C12H20Cl6 because of slower degradation rates and lower water solubility. Environmental residence time of C16H24Cl10 is estimated to be 520 d compared to 210 d for C12H20Cl6. Future studies will require better analytical methods and reference materials certified for PCA content. Additional data are needed to evaluate exposure of biota to PCAs in the environment, particularly in light of their continued production and usage around the globe. PMID- 9751034 TI - Brussels seeks BSE diagnostic test to screen European cattle. PMID- 9751036 TI - Biology teaching wins $90m boost from Howard Hughes institute. PMID- 9751035 TI - Don't copy our clone, say Dolly scientists. PMID- 9751037 TI - Human genome deadline cut by two years. PMID- 9751038 TI - US scientists call for fairer animal funding. PMID- 9751039 TI - First shots fired in biological warfare. PMID- 9751040 TI - Worm holes and avian space-time. PMID- 9751041 TI - Bacterial infection. For whom the bell tolls. PMID- 9751042 TI - Purposeful mutations. PMID- 9751043 TI - Ribozymes. How to make a nucleotide. PMID- 9751044 TI - Signal transduction. Docking IkappaB kinases. PMID- 9751045 TI - Mammary models. PMID- 9751046 TI - Mothers determine sexual preferences. PMID- 9751047 TI - All limbs are not the same. PMID- 9751048 TI - Does vitamin C have a pro-oxidant effect? PMID- 9751049 TI - Does vitamin C have a pro-oxidant effect? PMID- 9751050 TI - Structural basis for inhibition of the cyclin-dependent kinase Cdk6 by the tumour suppressor p16INK4a. AB - The cyclin-dependent kinases 4 and 6 (Cdk4/6) that control the G1 phase of the cell cycle and their inhibitor, the p16INK4a tumour suppressor, have a central role in cell proliferation and in tumorigenesis. The structures of Cdk6 bound to p16INK4a and to the related p19INK4d reveal that the INK4 inhibitors bind next to the ATP-binding site of the catalytic cleft, opposite where the activating cyclin subunit binds. They prevent cyclin binding indirectly by causing structural changes that propagate to the cyclin-binding site. The INK4 inhibitors also distort the kinase catalytic cleft and interfere with ATP binding, which explains how they can inhibit the preassembled Cdk4/6-cyclin D complexes as well. Tumour derived mutations in INK4a and Cdk4 map to interface contacts, solidifying the role of CDK binding and inhibition in the tumour suppressor activity of p16INK4a. PMID- 9751051 TI - Crystal structure of the complex of the cyclin D-dependent kinase Cdk6 bound to the cell-cycle inhibitor p19INK4d. AB - The crystal structure of the cyclin D-dependent kinase Cdk6 bound to the p19 INK4d protein has been determined at 1.9 A resolution. The results provide the first structural information for a cyclin D-dependent protein kinase and show how the INK4 family of CDK inhibitors bind. The structure indicates that the conformational changes induced by p19INK4d inhibit both productive binding of ATP and the cyclin-induced rearrangement of the kinase from an inactive to an active conformation. The structure also shows how binding of an INK4 inhibitor would prevent binding of p27Kip1, resulting in its redistribution to other CDKs. Identification of the critical residues involved in the interaction explains how mutations in Cdk4 and p16INK4a result in loss of kinase inhibition and cancer. PMID- 9751053 TI - Episodic-like memory during cache recovery by scrub jays. AB - The recollection of past experiences allows us to recall what a particular event was, and where and when it occurred, a form of memory that is thought to be unique to humans. It is known, however, that food-storing birds remember the spatial location and contents of their caches. Furthermore, food-storing animals adapt their caching and recovery strategies to the perishability of food stores, which suggests that they are sensitive to temporal factors. Here we show that scrub jays (Aphelocoma coerulescens) remember 'when' food items are stored by allowing them to recover perishable 'wax worms' (wax-moth larvae) and non perishable peanuts which they had previously cached in visuospatially distinct sites. Jays searched preferentially for fresh wax worms, their favoured food, when allowed to recover them shortly after caching. However, they rapidly learned to avoid searching for worms after a longer interval during which the worms had decayed. The recovery preference of jays demonstrates memory of where and when particular food items were cached, thereby fulfilling the behavioural criteria for episodic-like memory in non-human animals. PMID- 9751052 TI - RNA-catalysed nucleotide synthesis. AB - The 'RNA world' hypothesis proposes that early life developed by making use of RNA molecules, rather than proteins, to catalyse the synthesis of important biological molecules. It is thought, however, that the nucleotides constituting RNA were scarce on early Earth. RNA-based life must therefore have acquired the ability to synthesize RNA nucleotides from simpler and more readily available precursors, such as sugars and bases. Plausible prebiotic synthesis routes have been proposed for sugars, sugar phosphates and the four RNA bases, but the coupling of these molecules into nucleotides, specifically pyrimidine nucleotides, poses a challenge to the RNA world hypothesis. Here we report the application of in vitro selection to isolate RNA molecules that catalyse the synthesis of a pyrimidine nucleotide at their 3' terminus. The finding that RNA can catalyse this type of reaction, which is modelled after pyrimidine synthesis in contemporary metabolism, supports the idea of an RNA world that included nucleotide synthesis and other metabolic pathways mediated by ribozymes. PMID- 9751054 TI - Primary motor cortex is involved in bimanual coordination. AB - Many voluntary movements involve coordination between the limbs. However, there have been very few attempts to study the neuronal mechanisms that mediate this coordination. Here we have studied the activity of cortical neurons while monkeys performed tasks that required coordination between the two arms. We found that most neurons in the primary motor cortex (MI) show activity specific to bimanual movements (bimanual-related activity), which is strikingly different from the activity of the same neurons during unimanual movements. Moreover, units in the supplementary motor area (SMA; the area of cortex most often associated with bimanual coordination) showed no more bimanual-related activity than units in MI. Our results challenge the classic view that MI controls the contralateral (opposite) side of the body and that SMA is responsible for the coordination of the arms. Rather, our data suggest that both cortical areas share the control of bilateral coordination. PMID- 9751055 TI - Early-blind human subjects localize sound sources better than sighted subjects. AB - Do blind persons develop capacities of their remaining senses that exceed those of sighted individuals? Besides anecdotal suggestions, two views based on experimental studies have been advanced. The first proposes that blind individuals should be severely impaired, given that vision is essential to develop spatial concepts. The second suggests that compensation occurs through the remaining senses, allowing them to develop an accurate concept of space. Here we investigate how an ecologically critical function, namely three-dimensional spatial mapping, is carried out by early-blind individuals with or without residual vision. Subjects were tested under monaural and binaural listening conditions. We find that early-blind subjects can map the auditory environment with equal or better accuracy than sighted subjects. Furthermore, unlike sighted subjects, they can correctly localize sounds monaurally. Surprisingly, blind individuals with residual peripheral vision localized sounds less precisely than sighted or totally blind subjects, confirming that compensation varies according to the aetiology and extent of blindness. Our results resolve a long-standing controversy in that they provide behavioural evidence that totally blind individuals have better auditory ability than sighted subjects, enabling them to compensate for their loss of vision. PMID- 9751056 TI - Impaired febrile response in mice lacking the prostaglandin E receptor subtype EP3. AB - Fever, a hallmark of disease, is elicited by exogenous pyrogens, that is, cellular components, such as lipopolysaccharide (LPS), of infectious organisms, as well as by non-infectious inflammatory insults. Both stimulate the production of cytokines, such as interleukin (IL)-1beta, that act on the brain as endogenous pyrogens. Fever can be suppressed by aspirin-like anti-inflammatory drugs. As these drugs share the ability to inhibit prostaglandin biosynthesis, it is thought that a prostaglandin is important in fever generation. Prostaglandin E2 (PGE2) may be a neural mediator of fever, but this has been much debated. PGE2 acts by interacting with four subtypes of PGE receptor, the EP1, EP2, EP3 and EP4 receptors. Here we generate mice lacking each of these receptors by homologous recombination. Only mice lacking the EP3 receptor fail to show a febrile response to PGE2 and to either IL-1beta or LPS. Our results establish that PGE2 mediates fever generation in response to both exogenous and endogenous pyrogens by acting at the EP3 receptor. PMID- 9751057 TI - Toll-like receptor-2 mediates lipopolysaccharide-induced cellular signalling. AB - Vertebrates and invertebrates initiate a series of defence mechanisms following infection by Gram-negative bacteria by sensing the presence of lipopolysaccharide (LPS), a major component of the cell wall of the invading pathogen. In humans, monocytes and macrophages respond to LPS by inducing the expression of cytokines, cell-adhesion proteins, and enzymes involved in the production of small proinflammatory mediators. Under pathophysiological conditions, LPS exposure can lead to an often fatal syndrome known as septic shock. Sensitive responses of myeloid cells to LPS require a plasma protein called LPS-binding protein and the glycosylphosphatidylinositol-anchored membrane protein CD14. However, the mechanism by which the LPS signal is transduced across the plasma membrane remains unknown. Here we show that Toll-like receptor 2 (TLR2) is a signalling receptor that is activated by LPS in a response that depends on LPS-binding protein and is enhanced by CD14. A region in the intracellular domain of TLR2 with homology to a portion of the interleukin (IL)-1 receptor that is implicated in the activation of the IL-1-receptor-associated kinase is required for this response. Our results indicate that TLR2 is a direct mediator of signalling by LPS. PMID- 9751058 TI - Amino-acid transport by heterodimers of 4F2hc/CD98 and members of a permease family. AB - Amino-acid transport across cellular plasma membranes depends on several parallel functioning (co-)transporters and exchangers. The widespread transport system L accounts for a sodium-independent exchange of large, neutral amino acids, whereas the system y(+)L exchanges positively charged amino acids and/or neutral amino acids together with sodium. The molecular nature of these transporters remains unknown, although expression of the human cell-surface glycoprotein 4F2 heavy chain (h4F2hc; CD98 in the mouse) is known to induce low levels of L- and/or y(+)L-type transport. This glycoprotein is found in activated lymphocytes, together with an uncharacterized, disulphide-linked lipophilic light chain with an apparent relative molecular mass of 40,000 (M(r) 40K). Here we identify the permease-related protein E16 as the first light chain of h4F2hc and show that the resulting heterodimeric complex mediates L-type amino-acid transport. The homologous protein from Schistosoma mansoni, SPRM1, also associates covalently with coexpressed h4F2hc glycoprotein, although it induces amino-acid transport of different substrate specificity. The coexpression of h4F2hc is required for surface expression of these permease-related light chains, which belong to a new family of amino-acid transporters that form heterodimers with cell-surface glycoproteins. PMID- 9751059 TI - IKAP is a scaffold protein of the IkappaB kinase complex. AB - The transcription factor NF-kappaB coordinates the activation of numerous genes in response to pathogens and pro-inflammatory cytokines, and is, therefore, vital in the development of acute and chronic inflammatory diseases. NF-kappaB is activated by phsophorylation of its inhibitory subunit, IkappaB-alpha, on serine residues 32 and 36 by cytokine-activated IKB kinases (IKKs); this phosphorylation precedes rapid degradation of IkappaB. IKK-alpha and IKK-beta isozymes are found in large complexes of relative molecular mass 700,000-900,000 (M(r) 70K-90K), but little is known about other components that organize and regulate these complexes. IKK-alpha was independently discovered as a NF-kappaB-inducing kinase (NIK)-associated protein in a yeast two-hybrid screen, and IKK-beta was also identified by homology screening. It is, however, unknown whether NIK is part of the IKK complex. Here we isolate large, interleukin-1-inducible IKK complexes that contain NIK, IKK-alpha, IKK-beta, IkappaB-alpha, NF-kappaB/RelA and a protein of M(r) 150K. This latter component is a new protein, termed IKK-complex associated protein (IKAP), which can bind NIK and IKKs and assemble them into an active kinase complex. We show that IKAP is a scaffold protein and a regulator for three different kinases involved in pro-inflammatory cytokine signalling. PMID- 9751060 TI - IKK-gamma is an essential regulatory subunit of the IkappaB kinase complex. AB - Pro-inflammatory cytokines activate the transcription factor NF-kappaB by stimulating the activity of a protein kinase that phosphorylates IkappaB, an inhibitor of NF-kappaB, at sites that trigger its ubiquitination and degradation. This results in the nuclear translocation of freed NF-kappaB dimers and the activation of transcription of target genes. Many of these target genes code for immunoregulatory proteins. A large, cytokine-responsive IkappaB kinase (IKK) complex has been purified and the genes encoding two of its subunits have been cloned. These subunits, IKK-alpha and IKK-beta, are protein kinases whose function is needed for NF-kappaB activation by pro-inflammatory stimuli. Here, by using a monoclonal antibody against IKK-alpha, we purify the IKK complex to homogeneity from human cell lines. We find that IKK is composed of similar amounts of IKK-alpha, IKK-beta and two other polypeptides, for which we obtained partial sequences. These polypeptides are differentially processed forms of a third subunit, IKK-gamma. Molecular cloning and sequencing indicate that IKK gamma is composed of several potential coiled-coil motifs. IKK-gamma interacts preferentially with IKK-beta and is required for the activation of the IKK complex. An IKK-gamma carboxy-terminal truncation mutant that still binds IKK beta blocks the activation of IKK and NF-kappaB. PMID- 9751062 TI - The complexity of treating asthma. PMID- 9751061 TI - Signal-specific co-activator domain requirements for Pit-1 activation. AB - POU-domain proteins, such as the pituitary-specific factor Pit-1, are members of the homeodomain family of proteins which are important in development and homeostasis, acting constitutively or in response to signal-transduction pathways to either repress or activate the expression of specific genes. Here we show that whereas homeodomain-containing repressors such as Rpx2 seem to recruit only a co repressor complex, the activity of Pit-1 is determined by a regulated balance between a co-repressor complex that contains N-CoR/SMRT, mSin3A/B and histone deacetylases, and a co-activator complex that includes the CREB-binding protein (CBP) and p/CAF. Activation of Pit-1 by cyclic AMP or growth factors depends on distinct amino- and carboxy-terminal domains of CBP, respectively. Furthermore, the histone acetyltransferase functions of CBP or p/CAF are required for Pit-1 function that is stimulated by cyclic AMP or growth factors, respectively. These data show that there is a switch in specific requirements for histone acetyltransferases and CBP domains in mediating the effects of different signal transduction pathways on specific DNA-bound transcription factors. PMID- 9751063 TI - The complexity of treatment adherence in adults with asthma: challenges and opportunities. AB - The therapeutic program for persons with asthma includes recommendations for altering the environment and a drug regimen designed to alleviate symptoms, minimize exacerbations, and improve quality of life. Unfortunately, patients can have difficulty adhering to these recommendations, which contributes to treatment failure and increased costs. This paper provides a comprehensive review of the challenge of adherence in adults with asthma, including the costs and benefits, optimal adherence levels, assessment methods commonly used in research and practice, factors believed to predict poor adherence, and tested and untested strategies for improving adherence. Opportunities for further research are discussed throughout the paper. PMID- 9751064 TI - Exhaled nitric oxide and bronchial reactivity during and after inhaled beclomethasone in mild asthma. AB - The measurement of exhaled nitric oxide (ENO) is recognized as a marker of airway inflammation. ENO was measured in 10 nonsteroid-treated asthmatics at recruitment, during 3 weeks of inhaled beclomethasone (1000 microg/day) and for 3 weeks after withdrawal. Baseline ENO was increased in asthma compared with nonasthmatics (85.0+/-54.5 vs. 24.5+/-14.8 ppb, p < 0.0001). After inhaled steroid, there was no significant change in forced expiratory volume in 1 sec (FEV1) and forced vital capacity (FVC), but methacholine PC20 rose significantly (p = 0.0345). ENO (mean+/-SD; % baseline) fell after 1 week on steroid to 60.6+/ 31.1 and rose to 95.3+/-46.1 at 1 week after withdrawal. ENO did not correlate with PC20 or FEV1. The changes in ENO and PC20 were inversely correlated (r2 = 0.325). ENO may be an index of airway inflammation and therapeutic response in bronchial asthma. PMID- 9751065 TI - Anaerobic exercise testing in children with asthma. AB - Twenty-two males with asthma and 22 healthy males (7-18 years old) performed a Wingate Anaerobic Test. The subjects with asthma had a lower mid-maximal expiratory flow rate (78.3+/-25.3 vs. 100.1+/-24.5%, p < 0.01) than the controls. Aerobic and anaerobic exercise performance (peak power: PP; mean power: MP) were similar for the two groups whether expressed in absolute or relative terms to body weight. The subgroup with asthma with a lower percent body fat (<25%) had a higher PP and MP than the subjects with a higher percent body fat (>25%). Pulmonary function or maturation were not independent correlates of anaerobic power. Our results fail to show differences in anaerobic exercise performance in children with asthma compared to healthy controls. Nutrition appears to be an important factor for performance during this test. PMID- 9751066 TI - Blood eosinophilia and degree of sensitization to house dust mites in preschool and school children with asthma. AB - In allergic asthma, there is convincing evidence that changes in eosinophil and lymphocyte state of activation in blood may reflect disease activity. We evaluated whether simple blood eosinophil or lymphocyte counts in atopic children with asthma could reflect the degree of allergic sensitization. Seventy-six asthmatic children, sensitized to house dust mites (HDM), in stable conditions at the time of the study, and 53 sex- and age-matched controls (CTR) were studied. As compared to CTR, allergic patients showed higher eosinophil numbers and percentages (p < 0.001) but similar lymphocyte numbers and proportions (p > 0.1). Both in CTR and in allergic patients, eosinophil counts did not correlate with lymphocyte counts (p > 0.05; each comparison) but positive correlations were observed between eosinophil numbers and percentages and paper radio immunosorbent test (PRIST) levels or radio-allergo sorbent test (RAST) classes (p < 0.001; each comparison). When allergic asthmatic individuals were subdivided according to their age into two subgroups (Gr), no differences were found in eosinophil and lymphocyte counts and in PRIST levels and RAST values between Gr1 (< or =5 years old [preschool children]) and Gr2 (>5 years old [school children]) (p > 0.05; each comparison). Interestingly, although positive correlations between eosinophil counts and PRIST levels were found in both subgroups (p < 0.05; each comparison), only in Gr2 did eosinophil counts correlate positively with RAST classes (p < 0.001). No correlations between lymphocyte counts and PRIST levels or RAST classes were demonstrated (p > 0.05; each comparison). These data suggest that although blood eosinophilia was similar in preschool and in allergic asthmatic school children sensitized to HDM, only in the oldest children did blood eosinophil counts appear to be related to the degree of HDM-specific sensitization. PMID- 9751067 TI - Relationships among respiratory infections, triggers of attacks, and asthma severity in children. AB - The present study of asthmatic children examined relationships among the frequencies of prior respiratory infections (i.e., those prior to the development of asthma) and recent (past year) respiratory infections, asthma severity, and the impacts of 12 common asthma triggers: air pollution, allergy problems, anger, cigarette smoke, excitement, high humidity, high or low environmental temperature, laughter, nighttime hours, physical activity, respiratory infection, and stress or worry. Data on these variables were obtained through a survey in which 325 families completed questionnaires; 121 families had asthmatic children who were 2-20 years of age. Pearson correlational analyses revealed many significant positive correlations: The frequencies of prior and recent infections were correlated. The frequency of prior infections was correlated with the impacts of all asthma triggers except allergy problems, but the frequency of recent infections was correlated only with the impacts of air pollution, cigarette smoke, respiratory infection, and nighttime hours as triggers of asthma attacks. Asthma severity was correlated with the frequencies of prior and recent respiratory infections and with the impact of respiratory infection as an asthma trigger. PMID- 9751068 TI - Bronchodilator responses to salbutamol using diskhaler versus metered-dose inhaler. AB - In adults inhaling salbutamol via metered-dose inhalers (MDls) 200 microg doses are recommended, but with diskhalers the manufacturer advocates 400 rather than 200 microg doses. To assess this advice, a partially double-blind, placebo controlled salbutamol dose response, crossover study (also incorporating MDI doses) was conducted in 12 mild/moderate asthmatics. After active treatment, mean peak expiratory flow rate (PEFR) increments yielded no clinically or statistically significant differences; compared to placebo, respective median differences in PEFR increments (95% Cls) were 10 (-10, 50), 20 (0, 50), and 15 (0, 30) following 400 and 200 microg via diskhalers and 200 microg via MDls. Diskhalers are a suitable alternative for patients with poor MDI technique, but the use of 400 rather than 200 microg salbutamol doses is not supported by evidence. PMID- 9751069 TI - Assessment of the reliability, validity, and responsiveness of a Spanish Asthma Quality of Life questionnaire. AB - A Spanish-language questionnaire designed for measuring the impact of asthma on quality of life in adults was developed. It was derived, by the application of a rigorous translation protocol, from a previously validated, English-language Asthma Quality of Life (AQL) questionnaire which had been developed in Australia. The aim of this study was to evaluate the psychometric properties of the Spanish AQL questionnaire using a cross-sectional and longitudinal design. Two hundred ninety-four clinically stable subjects with asthma (168 women, mean baseline forced expiratory volume in 1 sec [FEV1] = 85% predicted), aged 17-70, attended for the initial baseline assessment. All subjects completed the AQL questionnaire and a full history and physical examination were performed. The clinical assessment of severity was based on the classification recommended by the Global Initiative on Asthma (GINA). One week after the initial assessment subjects completed the AQL questionnaire for a second time. Six months later, subjects were assessed clinically and completed all the assessment measures at baseline. Principal components analysis of the AQL questionnaire responses at the baseline visit revealed a structure that was almost identical to that seen in the original English-language questionnaire. The questionnaire was shown to be internally consistent (Cronbach's alpha 0.91 for total score and 0.80-0.86 for the four subscales) and repeatable (intraclass correlation coefficient 0.91 for the total scale and 0.78-0.92 for the subscales). The finding of expected strong correlations with the subject's global assessment of severity (p = 0.70) and dyspnea (p = 0.63), a weak inverse correlation with FEV1 (p = -0.17), and good discrimination among the four GINA severity categories (F3,291 = 37.16, p < 0.0001) supports the construct validity of the questionnaire. AQL scores increased with age (p = 0.31) and were higher in women (p < 0.005). The AQL was responsive to both improvement (mean change 1.02, p < 0.0001) and deterioration (mean change -1.13, p < 0.001) in the severity of asthma over a 6-month period. This disease-specific, Spanish-language AQL questionnaire was shown to have sound psychometric properties which make it suitable for use in cross-sectional or longitudinal studies where it is appropriate to assess the impact of asthma on the quality of life of individual patients. PMID- 9751070 TI - On the role of the peroxisome in the metabolism of drugs and xenobiotics. AB - One of the most rapidly developing areas of organellar biology, and one with major involvements in biochemical pharmacology, is that of peroxisomal function. In this commentary, several recent research findings in this area are described, along with their significance in relation to the metabolism of drugs and xenobiotics. Topics that are covered include the peroxisome proliferation caused by a wide variety of chemical compounds, the metabolic implications of this phenomenon, interactions between peroxisomal transcription factors and the eicosanoids, and the close relationships between peroxisomal metabolism and the control of functional damage caused by oxygen free radicals. Examples are also provided on interactions between peroxisomal function and such diverse drug groupings as analgesics, anti-inflammatories, nutritional ingestants, insecticides, antifungals, herbicides, and plasticisers. It is concluded that the peroxisome plays a central role in the metabolism of many drugs, displays a significant potential in furthering an understanding of pharmacological mechanisms, and is deserving of greater emphasis in future research considerations in this area. PMID- 9751071 TI - The Hsp90 complex--a super-chaperone machine as a novel drug target. AB - Cells respond to sudden changes in the environmental temperature with increased synthesis of a distinct number of heat shock proteins (Hsps). Analysis of the function of these proteins in recent years has shown that all the major classes of conserved Hsps are molecular chaperones involved in assisting cellular protein folding and preventing irreversible side-reactions, such as unspecific aggregation. In addition to their function under stress conditions, molecular chaperones also play a critical role under physiological conditions. Hsp90 is one of the most abundant chaperones in the cytosol of eukaryotic cells. It is part of the cell's powerful network of chaperones to fight the deleterious consequences of protein unfolding caused by nonphysiological conditions. In the absence of stress, however, Hsp90 is an obligate component of fundamental cellular processes such as hormone signaling and cell cycle control. In this context, several key regulatory proteins, such as steroid receptors, cell cycle kinases, and p53, have been identified as substrates of Hsp90. Recently, Hsp90 was shown to be the unique target for geldanamycin, a potent new anti-tumor drug that blocks cell proliferation. Interestingly, under physiological conditions, Hsp90 seems to perform its chaperone function in a complex with a set of partner proteins, suggesting that the Hsp90 complex is a multi-chaperone machine specialized in guiding the maturation of conformationally labile proteins. The regulation of key signaling molecules of the cell by the Hsp90 machinery is a stimulating new concept emerging from these studies, and Hsp90 has become a promising new drug target. PMID- 9751072 TI - Bruton's tyrosine kinase (BTK) as a dual-function regulator of apoptosis. AB - Multiple counterregulatory mechanisms have been identified in B-cell precursors that operate to regulate cell survival and growth, thereby ensuring the orderly development and differentiation of B-cells. Inappropriate apoptosis may underlie the pathogenesis of immunodeficiencies, as well as pathogenesis and drug/radiation resistance of human leukemias and lymphomas, which makes control of apoptosis an important potential target for therapeutic interventions. Therefore, identification of the molecular regulators of apoptosis is an area of intense investigation. Bruton's tyrosine kinase (BTK) is the first tyrosine kinase to be identified as a dual-function regulator of apoptosis, which promotes radiation-induced apoptosis but inhibits Fas-activated apoptosis in B-cells. BTK functions in a pro-apoptotic manner when B-cells are exposed to reactive oxygen intermediates, at least in part, by down-regulating the anti-apoptotic activity of STAT-3 transcription factor. In contrast, BTK associates with the death receptor Fas and impairs its interaction with Fas-associated protein with death domain (FADD), which is essential for the recruitment and activation of FLICE by Fas during the apoptotic signal, thereby preventing the assembly of a pro apoptotic death inducing signaling complex (DISC) after Fas-ligation. The identification of BTK as a dual-function regulator of apoptosis will significantly increase our understanding of both the biological processes involved in programmed cell death and the diseases associated with dysregulation of apoptosis. New agents with BTK-modulatory activity may have clinical potential in the treatment of B-cell malignancies (in particular acute lymphoblastic leukemia, the most common form of childhood cancer), as well as B-cell immunodeficiencies. PMID- 9751073 TI - Effects of oxatomide and derivatives on high affinity IgE receptor-activated signal transduction pathways in rat basophilic leukemia cells: role of protein tyrosine hyperphosphorylation and inhibition of extracellular calcium influx. AB - The antiallergic drug oxatomide and analogs inhibit mediator release from a rat basophilic leukemia (RBL-2H3) cell line, which is frequently used as a mast cell model. By investigating a series of derivatives of oxatomide with different inhibiting activities on exocytosis, we aimed to evaluate the role of their effects on the early steps of the signal transduction cascade in the inhibition of exocytosis. The active compounds induced hyperphosphorylation of tyrosine residues both in stimulated as well as in resting cells. Furthermore, some elevation of the inositol 1,4,5-trisphosphate (IP3) formation upon antigen activation was observed for the active derivatives. Ca2+ fluxes were also studied. The inhibition of the antigen-induced 45Ca2+ influx correlated with the effects of the drugs on exocytosis. Furthermore, the inhibitory activity on antigen- and thapsigargin-mediated exocytosis correlated well. Adherence of the cells to fibronectin, stimulating cellular integrin receptors, was synergistic to antigen activation of the RBL cells. However, oxatomide did lack any effect on integrin-mediated processes, as the IC50 value for exocytosis was identical for fibronectin-adhered cells and standard cultured cells. We conclude that oxatomide and its analogs inhibit exocytosis, mainly by inhibiting Ca2+ influx over store operated Ca2+ (SOC) channels. The drugs have a direct effect on the store operated Ca2+ channels or affect the direct regulation of these channels. PMID- 9751074 TI - In vitro stability of polymerase chain reaction-generated DNA fragments in serum and cell extracts. AB - The potential use of polymerase chain reaction (PCR)-generated DNA fragments (PCR DNAs) as pharmaceutical agents has previously been suggested, with the demonstration of the in vitro cellular internalization and biologic activity of PCR-DNA decoy molecules targeted to human estrogen receptor gene. In order to provide information on the stability of these double-stranded DNA molecules, the nuclease resistance of PCR-DNAs of different sizes was studied in different conditions and experiments. Simulating in vitro and in vivo transfection protocol, we demonstrated that PCR-DNAs exhibited good stability toward fetal bovine serum (FBS) and adult human serum nuclease digestion. In addition, when the protective activity of liposome-based formulations toward nuclease digestion was tested, it was shown that the stability of PCR-DNAs could be further increased (up to 7 days) when a liposome-mediated delivery system was employed. PMID- 9751075 TI - Phorbol ester-induced P-glycoprotein phosphorylation and functionality in the HTB 123 human breast cancer cell line. AB - The discordance between P-glycoprotein (P-gp) expression and functionality [as measured by the efflux of doxorubicin (DOX)] was analyzed in a DOX-sensitive human breast cancer cell line (HTB-123) with high reactivity against four P-gp specific monoclonal antibodies (C219, MRK-16, UIC2, and 4E3). Reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting analyses confirmed the overexpression of MDR1 mRNA and P-gp in this cell line. However, incubation of cells with efflux blockers, verapamil (VPL) or dipyridamole (DPD), did not enhance cellular (DOX) accumulation or cytotoxicity. Upon incubation with 12-O-tetradecanoylphorbol-13-acetate (TPA), HTB-123 cells retained less DOX than control cells and were sensitive to the efflux blockers verapamil or dipyridamole. These observations suggest that 12-O-tetradecanoylphorbol-13 acetate-induced P-gp phosphorylation may be associated with induction of P-gp mediated drug efflux in the HTB-123 cell line. PMID- 9751076 TI - Effect of modulators on the ATPase activity and vanadate nucleotide trapping of human P-glycoprotein. AB - P-Glycoprotein (Pgp) is responsible for the energy-dependent efflux of many natural product oncolytics. Overexpression of Pgp may result in multidrug resistance (MDR). Modulators can block Pgp efflux and sensitize multidrug resistant cells to these oncolytics. To study the interaction of modulators with Pgp, Pgp-ATPase activity was examined, using plasma membranes isolated from the multidrug-resistant cell line CEM/VLB100. A survey of modulators indicated that verapamil, trifluoperazine, and nicardipine stimulated ATPase activity by 1.3- to 1.8-fold, whereas two others, trimethoxybenzoylyohimbine (TMBY) and vindoline, had no effect. Further evaluation showed that TMBY completely blocked the stimulation by verapamil of ATPase activity by competitive inhibition, with a Ki of 2.1 microM. When the effects of these two modulators on the formation of the enzyme-nucleotide complex important in the catalytic cycle were examined, verapamil increased the amount of vanadate-trapped 8-azido-[alpha-32P]ATP bound to Pgp by two-fold, whereas TMBY had no effect. Moreover, TMBY blocked the verapamil stimulation of vanadate-8-azido-[alpha-32P]ATP. Together, these data indicate that verapamil and TMBY bind to Pgp at a common site or overlapping sites, but only verapamil results in enhanced Pgp-ATP hydrolysis and formation of the vanadate-nucleotide-enzyme complex. PMID- 9751077 TI - Differential effects of thiocyanate on the binding thermodynamics of bicuculline methiodide versus SR95531 (gabazine) to the gamma-aminobutyric acid receptor ionophore complex. AB - The temperature dependence of the binding of [3H]SR 95531 (Gabazine), an antagonist of gamma-aminobutyric acid (GABA(A)) receptors, was studied in synaptosomal membranes of rat brain in the presence of 50 mM KSCN. The displacing potencies of the antagonists bicuculline methiodide and Gabazine were determined at five temperatures between 0 degrees and 37 degrees. Van't Hoff plots of the displacing potencies were analyzed by linear regression in the presence and absence of thiocyanate. Thiocyanate hardly affected the exothermic ionic binding interaction of gabazine. In contrast, thiocyanate strongly potentiated the binding of bicuculline methiodide and deprived it of its exothermic nature. The enhanced binding of bicuculline methiodide in the presence of chaotropic SCN- ions might be reconciled with "entropic trapping" in a sterically constrained hydrophobic binding pocket. PMID- 9751078 TI - Direct binding of chloroquine to the multidrug resistance protein (MRP): possible role for MRP in chloroquine drug transport and resistance in tumor cells. AB - Multidrug resistance protein (MRP) transports a range of compounds that include glutathione S-conjugates, amphiphilic anionic drugs, and natural-product toxins. However, the mechanism of MRP drug binding and transport is presently unclear. We recently demonstrated the direct binding of a quinoline-based photoactive drug, N [4-[1-hydroxy-2-(dibutylamino)ethyl]quinolin-8-yl]-4-az idosalicylamide (IAAQ), to MRP at a biologically relevant site [Vezmar et al., Biochem Biophys Res Commun 241: 104-111, 1997]. In the present report, we demonstrated that the lysosomotropic or antimalarial drug chloroquine is a substrate for MRP. Specifically, our results showed that chloroquine, similar to leukotriene C4 (LTC4) and 3-(3-(2-(7-chloro-2-quinolinyl)ethenyl-phenyl)((3-(dimethyl amino-3 oxo propyl)thio)methyl)thio) propanoic acid (MK 571), inhibits the photoaffinity labeling of MRP by IAAQ. Furthermore, cell growth assays showed MRP-expressing multidrug-resistant cells (H69/AR and HL60/AR) to be more resistant to chloroquine than their parental cells (i.e., IC50 of 121 microM versus 28 microM chloroquine for H69/AR and H69, respectively). Moreover, MK 571, an LTD4 receptor antagonist, reversed the resistance of H69/AR cells to chloroquine. Drug transport studies using [14C]chloroquine demonstrated that MRP-expressing cells accumulate less drug than the parental drug-sensitive cells. The reduced accumulation of [14C]chloroquine in resistant cells was ATP dependent and was due to enhanced drug efflux. Taken together, the results of this study show that MRP modulates the transport of chloroquine by direct binding. PMID- 9751079 TI - Sensitization effect of L-2-oxothiazolidine-4-carboxylate on tumor cells to melphalan and the role of 5-oxo-L-prolinase in glutathione modulation in tumor cells. AB - 5-Oxo-L-prolinase (5-OPase) (EC 3.5.2.9) links the synthesis and metabolism of glutathione (GSH) in the gamma-glutamyl cycle. Previous studies showed that L-2 oxothiazolidine-4-carboxylate (OTZ), a 5-oxo-L-proline analog that is metabolized by 5-OPase, can preferentially decrease the cellular GSH levels in vivo in rat mammary tumors and sensitizes the tumors to the alkylating agent melphalan. The present study investigated the biochemical mechanism of this effect in a human breast cancer cell line, MCF7. We found that OTZ decreased the GSH levels in MCF7 cells. When the cells were treated with OTZ plus melphalan, the cytotoxicity of melphalan was increased as compared with that of melphalan alone, and this effect could be reversed by the addition of glutamate, which is the product of 5-OPase reaction and a critical substrate in GSH synthesis. We concluded that OTZ increases melphalan toxicity by limiting glutamate production from 5-OPase for GSH synthesis. We also observed that the expression of 5-OPase in the stably transfected MCF7 cells decreased the cellular GSH contents, sensitized the cells to melphalan toxicity, and diminished the sensitizing effect of OTZ. Furthermore, exposure to the GSH-depleting agent buthionine sulfoximine led to increased expression of 5-OPase in both MCF7 cells and the peripheral blood mononuclear cells of patients. These results indicate a critical interaction between cellular GSH levels and 5-OPase activity that could be important in GSH modulation in therapeutic settings. PMID- 9751080 TI - Effects of iron deprivation on the pathology and stress protein expression in murine X-linked muscular dystrophy. AB - Duchenne muscular dystrophy (DMD) is caused by dystrophin deficiency, which results in muscle necrosis and the upregulation of heat shock/stress proteins (HSP). We hypothesized that reactive oxygen species, and in particular hydroxyl radicals (.OH), participate in muscle necrosis and HSP expression. It was assumed that iron deprivation decreases .OH generation, restraining the disease process and reducing the oxidant-induced expression of HSP. The role of iron-catalyzed free radical reactions in the pathology of dystrophin-deficient muscle was evaluated in the murine model for Duchenne muscular dystrophy (mdx), by examining the effects of dietary deficiency and supplementation of iron on serum creatine kinase (CK), muscle morphology, lipid peroxidation and HSP levels in mice maintained on diets deficient in or supplemented with iron for 6 weeks. Iron deprived mdx mice showed a significant decrease in the number of macrophage invaded necrotic fibers and the expression of the 70-kDa heat shock protein (Hsp70). This suggests that the iron-dependent generation of .OH relates to muscle necrosis in the mdx mouse and modulates the expression of Hsp70 in vivo. In contrast, iron deprivation had no influence on other HSP or on lipid peroxidation in mdx mice, while maintenance on either diet caused a significant decrease in serum creatine kinase activity. The potential therapeutic effects of iron deprivation in mdx should be considered. PMID- 9751081 TI - Induction by interleukin-1beta peptide of prostaglandin E2 formation via enhanced prostaglandin H synthase-2 expression in 3T6 fibroblasts. AB - Several synthetic interleukin-1 (IL-1) peptides were tested in vivo for pyrogenic activity and in vivo for their ability to stimulate prostaglandin production. Only the IL-1beta fragment (208-240) enhanced body temperature, although both IL 1beta (208-240) and IL-1alpha (223-250) stimulated prostaglandin E2 (PGE2) production in vitro. We report here that the IL-1beta fragment (208-240) did not have the capacity to induce arachidonic acid (AA) mobilization by 3T6 fibroblasts. However, this peptide was able to increase the expression of the inducible prostaglandin H synthase isoform (PGHS-2; EC 1.14.99.1.), which is related to its ability to stimulate prostaglandin E2 synthesis. PMID- 9751082 TI - Hepatic mercapturic acid formation: involvement of cytosolic cysteinylglycine S conjugate dipeptidase activity. AB - The role of cysteinylglycine S-conjugate dipeptidases in the intrahepatic mercapturic acid pathway was investigated in rat liver. Subcellular compartmentation studies and liver perfusions were performed using monochlorobimane and bimane S-conjugates as model compounds. The major part (over 95%) of total hepatic cysteinylglycine S-conjugate dipeptidase activity was located in the cytosol. Lower specific activity appeared in the canalicular plasma membrane fraction. Similar hepatic localization of dipeptidase activity was seen in the guinea pig. In intact rat liver perfused with monochlorobimane, the major products were the glutathione S-conjugate (mBSG) and the cysteinylglycine S-conjugate (mBCG) in bile. Minor amounts of the cysteine S conjugate (mBCys) and the mercapturic acid (mBNAc) were formed, indicating a limitation in further metabolism of the dipeptide S-conjugate in the biliary space. However, when the dipeptide S-conjugate was offered to the sinusoidal space in liver perfusions, substantial uptake and conversion to mBNAc was observed, and only trace amounts of the infused dipeptide appeared in bile. The data suggest that cytosolic cysteinylglycine S-conjugate dipeptidase as identified here is involved in hepatic mercapturic acid formation from sinusoidal cysteinylglycine S-conjugates. This is especially of significance for species such as guinea pig and human, in which dipeptide S-conjugates are generated in the sinusoidal domain of the liver due to the presence of high gamma glutamyltranspeptidase activity. PMID- 9751083 TI - Nitric oxide protection of rat liver from lipid peroxidation, collagen accumulation, and liver damage induced by carbon tetrachloride. AB - The aim of this work was to determine if the inhibition or stimulation of NO synthesis modulates liver damage induced by the chronic administration of CCl4. CCl4 was administered three times a week for 8 weeks to male Wistar rats treated simultaneously with N omega-nitro-L-arginine methyl ester (L-NAME, 100 mg/kg, p.o., twice a day), aminoguanidine (AG, 4 g/L in the drinking water), or L arginine (500 mg/kg, p.o., twice a day); appropriate controls were performed. Serum NO2- + NO3- increased in the groups treated with CCl4 and/or L-arginine, but the effect was prevented by either L-NAME or AG. In the liver, lipid peroxidation and collagen content increased, while glycogen content decreased in the CCl4-treated group (P < 0.05); L-NAME and AG accentuated these effects. Serum enzyme activities of alanine aminotransferase (ALT), alkaline phosphatase, and gamma-glutamyl transpeptidase (gamma-GTP) and bilirubin content increased about 2 , 3-, 2-, and 6-fold, respectively, after CCl4 intoxication (P < 0.05); L-NAME or AG cotreatment further increased the enzyme activities (P < 0.05). L-Arginine treatment protected the liver partially from the elevation of collagen, bilirubins, and alkaline phosphatase and from glycogen depletion induced by CCl4 intoxication (P < 0.05), but showed no significant effect on ALT, gamma-GTP, or lipid peroxidation. These results suggest that NO protects the liver against oxidative injury, because NO inhibition by L-NAME or AG increased lipid peroxidation and the other markers of liver injury studied herein. PMID- 9751084 TI - Antineutrophil cytoplasmic antibodies. PMID- 9751085 TI - The immunopathogenesis and management of Kawasaki syndrome. PMID- 9751086 TI - Combination therapy with biologic agents in rheumatoid arthritis: perils and promise. PMID- 9751087 TI - Therapeutic efficacy of multiple intravenous infusions of anti-tumor necrosis factor alpha monoclonal antibody combined with low-dose weekly methotrexate in rheumatoid arthritis. AB - OBJECTIVE: To evaluate the efficacy, pharmacokinetics, immunogenicity, and safety of multiple infusions of a chimeric monoclonal anti-tumor necrosis factor alpha antibody (cA2) (infliximab; Remicade, Centocor, Malvern, PA) given alone or in combination with low-dose methotrexate (MTX) in rheumatoid arthritis (RA) patients. METHODS: In a 26-week, double-blind, placebo-controlled, multicenter trial, 101 patients with active RA exhibiting an incomplete response or flare of disease activity while receiving low-dose MTX were randomized to 1 of 7 groups of 14-15 patients each. The patients received either intravenous cA2 at 1, 3, or 10 mg/kg, with or without MTX 7.5 mg/week, or intravenous placebo plus MTX 7.5 mg/week at weeks 0, 2, 6, 10, and 14 and were followed up through week 26. RESULTS: Approximately 60% of patients receiving cA2 at 3 or 10 mg/kg with or without MTX achieved the 20% Paulus criteria for response to treatment, for a median duration of 10.4 to >18.1 weeks (P < 0.001 versus placebo). Patients receiving cA2 at 1 mg/kg without MTX became unresponsive to repeated infusions of cA2 (median duration 2.6 weeks; P=0.126 versus placebo). However, coadministration of cA2 at 1 mg/kg with MTX appeared to be synergistic, prolonging the duration of the 20% response in >60% of patients to a median of 16.5 weeks (P < 0.001 versus placebo; P=0.006 versus no MTX) and the 50% response to 12.2 weeks (P < 0.001 versus placebo; P=0.002 versus no MTX). Patients receiving placebo infusions plus suboptimal low-dose MTX continued to have active disease, with a Paulus response lasting a median of 0 weeks. A 70-90% reduction in the swollen joint count, tender joint count, and C-reactive protein level was maintained for the entire 26 weeks in patients receiving 10 mg/kg of cA2 with MTX. In general, treatment was well tolerated and stable blood levels of cA2 were achieved in all groups, except for the group receiving 1 mg/kg of cA2 alone, at which dosage antibodies to cA2 were observed in approximately 50% of the patients. CONCLUSION: Multiple infusions of cA2 were effective and well tolerated, with the best results occurring at 3 and 10 mg/kg either alone or in combination with MTX in approximately 60% of patients with active RA despite therapy with low-dose MTX. When cA2 at 1 mg/kg was given with low-dose MTX, synergy was observed. The results of the trial provide a strategy for further evaluation of the efficacy and safety of longer-term treatment with cA2. PMID- 9751088 TI - Should improvement in rheumatoid arthritis clinical trials be defined as fifty percent or seventy percent improvement in core set measures, rather than twenty percent? AB - OBJECTIVE: To determine whether improvement of more than 20% in core set parameters should be required before patients are characterized as improved in rheumatoid arthritis (RA) clinical trials. METHODS: Data from 6 RA trials were reanalyzed to evaluate the discriminant validity (ability to differentiate active treatment from control) of 4 proposed definitions of improvement: the current American College of Rheumatology (ACR) definition (a 20% threshold for core set parameters [ACR 201), a 50% threshold (ACR 50), a 70% threshold (ACR 70), and an ordinal definition in which a patient could be classified in any of 3 categories (unimproved, ACR 20, or ACR 50). To evaluate the discriminant validity of these 4 definitions of improvement, we characterized each patient in each trial as improved or not, based on each definition, and computed a chi-square value differentiating the active treatment group from the control group, with the corresponding P value. RESULTS: With an increase in the threshold for improvement, the percentage of placebo-treated patients who were classified as experiencing response dropped dramatically in all trials, as did the percentage of patients receiving active therapy (second-line drug, combination therapy, tumor necrosis factor p75-Fc fusion protein) who were classified as experiencing response. Generally, the drop in active treatment response rates was greater than the drop in placebo response rates, leaving the difference between the 2 groups less at the higher thresholds. Therefore, chi-square values fell as the threshold for response was raised. The ordinal definition of improvement yielded chi-square values similar to those obtained using ACR 20 alone. CONCLUSION: Adopting a definition of efficacy in RA trials that requires 50% or 70% improvement in core set parameters would likely compromise statistical power and make it more difficult to distinguish between 2 treatments with different efficacy. ACR 20 should continue to be the primary measure of efficacy in RA trials, with higher thresholds for improvement being determined and reported as secondary efficacy measures. PMID- 9751089 TI - Radiographic outcome of recent-onset rheumatoid arthritis: a 19-year study of radiographic progression. AB - OBJECTIVE: To describe the longitudinal radiographic course of rheumatoid arthritis (RA), and to identify and quantitate predictors of radiographic progression. METHODS: This prospective, longitudinal study of radiographic progression and clinical predictors of RA involved 256 patients with RA who were seen within the first 2 years of disease (mean 0.77 years) and were followed up for up to 19 years. Participants underwent a total of 6,278 clinical assessments (mean 24.5) and 934 paired radiographs (mean 3.1, range 2-6). Clinical assessments at every visit included determination of the erythrocyte sedimentation rate (ESR), grip strength, pain scores, tender joint counts, and anxiety and depression measurements. Regression analyses utilized time-integrated predictors. RESULTS: Overall, radiographic progression rates, as measured by the summary Sharp scores, appeared constant over the course of RA. The strongest correlate of progression was the time-integrated ESR (rho=0.53). This association grew stronger with time. At 0-5 years, 5-10 years, 10-15 years, and 15-20 years, correlations were 0.40, 0.50, 0.65, and 0.74, respectively, and for the period 10 20 years, the correlation was 0.67. In multivariate models, the mean ESR, mean grip strength, rheumatoid factor positivity, and tender joint count were independent predictors of radiographic progression. CONCLUSION: Radiographic damage occurs at a constant rate in RA, and is not greater early in RA or reduced later in the course of the illness. Acute-phase reactants are, by far, the strongest determinants of progression. PMID- 9751090 TI - Assessment of rheumatoid arthritis using a modified scoring method on digitized and original radiographs. AB - OBJECTIVE: The results of different readers' interpretations of laser-digitized hand radiographs versus original radiographs were compared to determine the reproducibility of scoring of erosions (ERO), joint space narrowing (JSN), and their combination (ERO + JSN) in patients with rheumatoid arthritis (RA). METHODS: Standardized radiographs of both hands were obtained at 2 visits (baseline and 6-24-month followup) from 30 patients with established RA. Conventional and laser-digitized (pixel sizes 50 microm and 100 microm) radiographs were scored independently by 3 experienced and trained radiologists who were blinded to the order of the visits. Scoring of radiographs was based on the validated Genant grading system. RESULTS: Intertechnique (intrareader) correlation coefficients at baseline were 0.90-0.93 for scoring of ERO, 0.90-0.94 for scoring of JSN, and 0.92-0.95 for ERO + JSN; for scoring of progression between baseline and followup, these values were 0.93-0.97, 0.87-0.95, and 0.93 0.97, respectively. Interreader (intratechnique) correlation coefficients at baseline were 0.82-0.96 for scoring of ERO, 0.69-0.91 for scoring of JSN, and 0.80-0.95 for ERO + JSN; for scoring of progression between baseline and followup, these values were 0.90-0.97, 0.80-0.92, and 0.90-0.95, respectively. Intrareader (intratechnique) correlation coefficients were 0.90-0.97 for scoring of the original radiographs and 0.90-0.98 for scoring of the digitized images at 100 microm. CONCLUSION: Using this modified grading system, scoring of RA progression directly from paired, high-resolution monitors of laser-digitized images of the hands provided highly reproducible results, comparable to those obtained from the original radiographs. Thus, this method may have useful applications in clinical trials involving RA. PMID- 9751091 TI - Preliminary study of the safety and efficacy of SC-58635, a novel cyclooxygenase 2 inhibitor: efficacy and safety in two placebo-controlled trials in osteoarthritis and rheumatoid arthritis, and studies of gastrointestinal and platelet effects. AB - OBJECTIVE: To investigate the efficacy and safety of SC-58635 (celecoxib), an antiinflammatory and analgesic agent that acts by selective cyclooxygenase 2 (COX 2) inhibition and is not expected to cause the typical gastrointestinal (GI), renal, and platelet-related side effects associated with inhibition of the COX-1 enzyme. METHODS: Four phase II trials were performed: a 2-week osteoarthritis efficacy trial, a 4-week rheumatoid arthritis efficacy trial, a 1-week endoscopic study of GI mucosal effects, and a 1-week study of effects on platelet function. RESULTS: The 2 arthritis trials identified SC-58635 dosage levels that were consistently effective in treating the signs and symptoms of arthritis and were distinguished from placebo on standard arthritis scales. In the upper GI endoscopy study, 19% of subjects receiving naproxen (6 of 32) developed gastric ulcers, whereas no ulcers occurred in subjects receiving SC-58635 or placebo. The study of platelet effects revealed no meaningful effect of SC-58635 on platelet aggregation or thromboxane B2 levels, whereas aspirin caused significant decreases in 2 of 3 platelet aggregation measures and thromboxane B2 levels. In all 4 trials, SC-58635 was well tolerated, with a safety profile similar to that of placebo. CONCLUSION: SC-58635 achieves analgesic and antiinflammatory efficacy in arthritis through selective COX-2 inhibition, without showing any evidence of 2 of the toxic effects of COX-1 inhibition associated with nonsteroidal antiinflammatory drugs. PMID- 9751092 TI - Codeine and oxycodone use in patients with chronic rheumatic disease pain. AB - OBJECTIVE: Opioid treatment of chronic rheumatic disease pain is controversial because of concerns regarding efficacy, toxicity, tolerance, dependence, and abuse. This study examined opioid use in a cohort of patients with pain due to defined rheumatic diseases. METHODS: Opioid use was studied retrospectively in a cohort of 644 rheumatology clinic patients. Computerized pharmacy records identified patients who had been prescribed opioids during the previous 3 years. Medical records were reviewed to determine reasons for opioid dosage escalations. Patients were interviewed to determine efficacy, frequency and types of side effects, and history of alcohol or street-drug abuse. RESULTS: Opioid prescriptions were found in the 3-year pharmacy database for 290 of 644 clinic patients: 153 for <3 consecutive months and 137 for > or =3 months. All opioid treated patients received codeine and/or oxycodone. In this cohort, 133 patients in each opioid-treated group and 76 of the 354 non-opioid-treated control patients were studied. Opioids significantly reduced rheumatic disease pain severity scores from 8.2 to 3.6 (on a 0-10 scale) (P < 0.001). Mild side effects were reported in 38%; nausea, dyspepsia, constipation, and sedation were the most common. The mean +/-SD initial dosage was 2.1+/-1.7 30-mg codeine equivalents/day, the mean peak was 3.4+/-3.3 per day, and the mean current dose was 2.7+/-2.0 per day. Dosage escalations occurred in 32 patients and were attributable to worsening of the underlying painful condition or a medical complication thereof in all but 4 patients, who also displayed other abuse behaviors. Abuse behaviors were not more frequent in those with or without a history of abuse/ addiction. CONCLUSION: Prolonged treatment of rheumatic disease pain with codeine or oxycodone was effective in reducing pain severity and was associated with only mild toxicity. Doses were stable for prolonged periods of time, with escalations of the opioid dose almost always related to worsening of the painful condition or a complication thereof, rather than the development of tolerance to opioids. Doubts or concerns about opioid efficacy, toxicity, tolerance, and abuse or addiction should no longer be used to justify withholding opioids from patients with well-defined rheumatic disease pain. PMID- 9751093 TI - Case-control study of corticosteroids and other drugs that either precipitate or protect from the development of scleroderma renal crisis. AB - OBJECTIVE: To determine whether the initiation of corticosteroids or other types of therapy affects the development of scleroderma renal crisis (SRC). METHODS: Using a case-control study, 110 patients with systemic sclerosis who developed SRC between 1981 and 1993 were closely matched with controls on sex, race, age, disease duration, skin score, levels of creatine phosphokinase, and presence of tendon friction rubs. Corticosteroid use was determined prior to the onset of SRC in cases or prior to the first visit in controls. Cases were compared with matched controls using McNemar's matched-pair analysis and conditional logistic regression analysis. The effects of other drugs, including D-penicillamine, nonsteroidal antiinflammatory drugs (NSAIDs), calcium channel blockers, and angiotensin-converting enzyme (ACE) inhibitors, were also evaluated. RESULTS: In the 6 months prior to SRC onset or to the first visit, high-dose corticosteroids (> or =15 mg/day prednisone or equivalent) were administered significantly more frequently in SRC patients (36%) than in the controls (12%) (McNemar's odds ratio 4.37, 95% confidence interval 2.03-9.43, P < 0.0001). New use of low-dose steroids, continuous use of any steroid dose, NSAIDs, calcium channel blockers, and ACE inhibitors were not associated with an increased risk of SRC. Antecedent D-penicillamine therapy may have been protective against the development of SRC in controls. CONCLUSION: This retrospective case-control study has shown a significant association between antecedent high-dose corticosteroid therapy and the development of SRC. These results should discourage the use of high-dose corticosteroids in patients with early diffuse scleroderma who are at increased risk of developing SRC. PMID- 9751094 TI - Transmission disequilibrium as a test of linkage and association between HLA alleles and pauciarticular-onset juvenile rheumatoid arthritis. AB - OBJECTIVE: To determine if HLA class I and II alleles previously found to be associated with (or protective against) pauciarticular-onset juvenile rheumatoid arthritis (pauci-onset JRA) in population-association studies are transmitted from heterozygous parents to an extent different from the expected 50%. METHODS: One hundred one Caucasian North American families that had a child with pauci onset JRA and at least 1 parent who was heterozygous for the allele of interest were available for analysis. Both biologic parents and all children (affected and unaffected) were typed for HLA class I and II alleles. The transmission disequilibrium test (TDT) was used to determine if affected offspring received the disease-associated (or protective) allele more (or less) frequently than its alternate allele. In families in which an unaffected sibling was available, the unmatched chi-square test was used to determine if a meiotic segregation distortion bias existed. RESULTS: HLA class I alleles A2, B27, and B35 showed a significantly higher than expected frequency of transmission to affected offspring, as did class II alleles DR5 and DR8. HLA-DR4 was found to be transmitted significantly less frequently than expected to affected, but not unaffected, offspring. All alleles that showed an excess transmission to affected offspring were transmitted to unaffected offspring at expected rates. When the data were stratified by age and sex, the likelihood of transmission of some of the alleles was strongly influenced by these variables. For example, excess transmission of HLA-DR5 was found exclusively in female patients who were younger at the time of disease onset. CONCLUSION: Results from these family-based studies rule out the possibility that HLA disease associations found in earlier studies were a result of population stratification and establish linkage and association between the major histocompatibility complex and pauci-onset JRA. PMID- 9751095 TI - Spectrum and clinical significance of autoantibodies against transfer RNA. AB - OBJECTIVE: To characterize the clinical features of patients who have autoantibodies against transfer RNA (tRNA) or tRNA-associated proteins. METHODS: Sera from 1,472 patients with suspected systemic rheumatic disease were screened by RNA immunoprecipitation of HeLa cell extracts. The specificities of the antibodies that precipitated tRNAs were further analyzed by immunoprecipitation using deproteinized RNAs and 35S-methionine-labeled HeLa cell extracts, followed by immunoblotting. RESULTS: Forty-one serum samples (2.8%) were found to immunoprecipitate tRNAs. Thirteen patients were identified as having previously defined anti-aminoacyl-tRNA synthetase antibodies (anti-histidyl-tRNA synthetase in 4 patients, anti-threonyl-tRNA synthetase in 1, anti-alanyl-tRNA synthetase in 3, anti-glycyl-tRNA synthetase in 4, and anti-isoleucyl-tRNA synthetase in 1). All 13 patients had myositis and/or interstitial pneumonitis. Sera from the remaining 28 patients immunoprecipitated previously unidentified tRNAs, including 13 serum samples that bound deproteinized cognate tRNA; 24 of the 28 patients met criteria for either systemic lupus erythematosus (SLE) or Sjogren's syndrome (SS). In addition, nonerosive polyarthritis, leukocytopenia, rheumatoid factor, and characteristic annular or papulosquamous recurrent erythema were noted in these patients; however, renal involvement was rare. Sera from 16 of these 28 patients also contained anti-Ro/SSA and/or anti-La/SSB antibodies. While 189 patient sera precipitated Ro/SSA and/or La/SSB-associated RNAs but not tRNA, only 12 of the patients (6.3%) developed skin lesions (P=0.0009, odds ratio 8.85). CONCLUSION: Novel autoantibodies against tRNAs or tRNA-associated proteins were identified in 28 sera. These autoantibodies appear to be distinct from anti aminoacyl-tRNA synthetase antibodies and are associated with SLE and SS. The presence of anti-Ro/SSA and/or anti-La/SSB along with anti-tRNA antibodies is more strongly associated with recurrent erythema than is the presence of anti Ro/SSA or anti-La/SSB alone. PMID- 9751096 TI - Linkage of chondrocyte apoptosis and cartilage degradation in human osteoarthritis. AB - OBJECTIVE: To examine the occurrence of apoptosis in human osteoarthritis (OA) cartilage, and to determine its relationship to cartilage degradation. METHODS: Knee cartilage was obtained from subjects at autopsy, from a tissue bank, and from OA patients undergoing total joint replacement surgery. Chondrocytes were isolated and the number of apoptotic cells was analyzed by flow cytometry. Apoptotic cells in cartilage sections were identified by the detection of DNA strand breaks. Electron microscopy was applied to demonstrate morphologic changes, and Safranin O staining was performed to analyze the relationship between apoptosis and proteoglycan depletion. RESULTS: Flow cytometry on cell suspensions prepared from collagenase digests of cartilage showed that approximately 22.3% of OA chondrocytes and 4.8% of normal chondrocytes were undergoing apoptosis. Staining of cartilage sections demonstrated the presence of apoptotic cells in the superficial and middle zones. Cartilage areas that contained apoptotic cells showed proteoglycan depletion, and the number of apoptotic cells was significantly correlated with the OA grade. CONCLUSION: These observations demonstrate increased chondrocyte apoptosis in OA cartilage. Chondrocyte apoptosis and proteoglycan depletion are anatomically linked and may be mechanistically related. PMID- 9751097 TI - Ro 32-3555, an orally active collagenase selective inhibitor, prevents structural damage in the STR/ORT mouse model of osteoarthritis. AB - OBJECTIVE: To determine the efficacy of a selective inhibitor of collagenases in an animal model of osteoarthritis (OA). METHODS: Ro 32-3555, an orally active collagenase selective inhibitor, was administered to STR/ORT mice. Microfocal x ray-generated images of the hind limbs were visually scored for joint space narrowing, osteophyte formation, and calcification of tendons. Histologic sections of the knees were scored for cartilage changes including loss of surface matrix, fibrillation, and eburnation. RESULTS: Significant inhibition of joint space narrowing and osteophyte formation was achieved in groups of animals treated with 10-50 mg/kg(-1) of Ro 32-3555. These effects were confirmed histologically in the same groups of animals: histologic analysis revealed that Ro 32-3555 protected cartilage from degradative changes. CONCLUSION: Ro 32-3555, a collagenase selective inhibitor, inhibits both the cartilage and bone changes in this mouse model of OA, and thus shows great potential as a treatment of OA in humans. PMID- 9751098 TI - Leukotriene A4 hydrolase and leukotriene C4 synthase activities in human chondrocytes: transcellular biosynthesis of Leukotrienes during granulocyte chondrocyte interaction. AB - OBJECTIVE: To investigate the cooperation of chondrocytes and polymorphonuclear cells (PMN) in the biosynthesis of leukotrienes (LT). METHODS: PMN, resting and interleukin-1beta-stimulated cultured human chondrocytes, and mixtures of both cell types were incubated with A23187 and/or 14C-arachidonic acid (14C-AA). To explore the presence of LTC4 synthase and LTA4 hydrolase, the chondrocytes were incubated with authentic LTA4. Eicosanoids were analyzed using high performance liquid chromatography techniques. RESULTS: Chondrocytes formed only prostaglandin E2 and minor amounts of 15-HETE and 11-HETE, the production of all of which was inhibited by 1 microM indomethacin. Incubation of PMN and chondrocytes produced more LTC4 from endogenous and exogenous AA, and more LTB4 from endogenous AA, than incubation of PMN alone, which was consistent with the presence of LTC4 synthase and LTA4 hydrolase activities in chondrocytes. Chondrocytes also slightly increased the level of PMN production of all 5-lipoxygenase (5-LO) derived products from endogenous AA. CONCLUSION: Human chondrocytes form eicosanoids from AA only by the cyclooxygenase pathway. Chondrocytes cooperate in the transcellular biosynthesis of LT since they possess LTA4 hydrolase and LTC4 synthase activities and increase metabolism by the 5-LO pathway in PMN. PMID- 9751099 TI - Accumulation of splenic B1a cells with potent antigen-presenting capability in NZM2410 lupus-prone mice. AB - OBJECTIVE: In order to shed light on the role of splenic B1 cells in disease pathogenesis in lupus-prone mice, this study was undertaken to determine how efficiently these cells can serve as antigen-presenting cells (APC) and to ascertain which murine lupus susceptibility loci dictate the expansion of these cells. METHODS: Spleens and peritoneal cavities (PerC) of NZM2410 lupus-prone mice, as well as of control B6 and New Zealand white mice, were examined for the prevalence, surface phenotype, and possible anatomic location of B1 cells. The antigen-presenting ability of fluorescence-sorted splenic B1a cells was assessed. Levels of B1 cells were examined in B6 mice congenic for 4 different lupus susceptibility intervals. RESULTS: NZM2410 lupus mice showed an expansion of splenic and PerC B1a cells at all ages. These cells expressed high levels of B71, B72, CD24, lymphocyte function-associated antigen 1, and intercellular adhesion molecule 1, and had the functional capability to serve as APC. Among the lupus susceptibility intervals studied, Sle2, but not Sle1, Sle3, or the H2 locus, affected the expansion of B1 cells. CONCLUSION: These findings raise the possibility that the genetically determined expansion of splenic B1a cells in lupus-prone mice might contribute to disease pathogenesis by augmenting the presentation of autoantigens to pathogenic T cells. PMID- 9751100 TI - Association of systemic lupus erythematosus with promoter polymorphisms of the mannose-binding lectin gene. AB - OBJECTIVE: To investigate the distribution of promoter variants of the mannose binding lectin (MBL) gene and correlations between the promoter variants and serum MBL concentrations in Chinese patients with systemic lupus erythematosus (SLE) and in healthy Chinese controls. METHODS: We studied the serum MBL levels and codon 54 mutation in 112 Chinese patients with SLE and 110 healthy controls. Genotyping of promoter variants of the MBL gene were done by polymerase chain reaction and allele-specific oligonucleotide hybridization. RESULTS: We found significant differences in the distribution of the 2 pairs of promoter polymorphisms, H/L and Y/X, between SLE patients and controls (P=0.018 and P=0.019, respectively). Analysis of the correlation between promoter haplotypes and serum MBL levels revealed HY as the highest-producing, LY as the intermediate producing, and LX as the lowest-producing haplotypes. The LX haplotype was present at a frequency of 0.259 in SLE patients and 0.154 in controls and was significantly associated with SLE (P=0.019, odds ratio 1.79, 95% confidence interval 1.12-2.85). CONCLUSION: The low-producing promoter polymorphism of the MBL gene is associated with SLE, and a low serum MBL level is a risk factor for SLE. Even allowing for promoter polymorphisms and structural mutations of the MBL gene, serum MBL levels in SLE patients are still lower than those in controls, suggesting a trans-factor in regulating serum MBL levels. PMID- 9751101 TI - Flow cytometric single-cell analysis of cytokine production by CD4+ T cells in synovial tissue and peripheral blood from patients with rheumatoid arthritis. AB - OBJECTIVE: To determine the cytokine profile of CD4+ T cells in the synovial tissue (ST) of rheumatoid arthritis (RA) patients at the single-cell level. METHODS: Unseparated ST cells and paired CD4+ T cells separated from the peripheral blood (PB) and ST of RA patients were stimulated for 4 hours with phorbol myristate acetate (PMA) plus calcium ionophore A23187, or for 6 hours with immobilized anti-CD3 plus anti-CD28, in the presence of brefeldin A. Cells were stained for intracellular cytokines such as interferon-gamma (IFNgamma), interleukin-2 (IL-2), IL-4, IL-10, and IL-13, in combination with cell surface markers. The percentages of cytokine-producing T cells were analyzed by flow cytometry. RESULTS: When ST cells were stimulated with PMA plus A23187 in bulk culture, IFNgamma-producing T cells were more frequently detected in the CD8+ subset, but cells producing other cytokines were found in the CD4+ subset. Purified ST CD4+ T cells, after stimulation with PMA plus A23187, were able to produce higher levels of IFNgamma but lower levels of IL-4 and IL-13, by analysis at the single-cell level, as compared with the PB CD4+, CD45RO+ T cells. The majority of IL-4- or IL-13-producing ST CD4+ cells produced IFNgamma, although PB CD4+ T cells rarely showed this cytokine pattern. IL-10-producing CD4+ T cells were more frequently found in the ST than in the PB. Of interest, most of the IL 10-producing ST CD4+ T cells were able to produce IFNgamma. IL-2-producing CD4+ T cells were similarly present in both compartments. Similar intracellular cytokine patterns were observed with anti-CD3 plus anti-CD28 stimulation, although the number of detected cells was lower. CONCLUSION: These data indicate that CD4+ T cells present within the inflamed synovium have apparently distinct cytokine profiles from those of memory CD4+ T cells in the PB, as typified by their ability to secrete both IFNgamma and IL-10. PMID- 9751102 TI - Interference of nonsteroidal antiinflammatory drugs with very late activation antigen 4/vascular cells adhesion molecule 1-mediated lymphocyte-endothelial cell adhesion. AB - OBJECTIVE: To study the effect of nonsteroidal antiinflammatory drugs (NSAIDs) on the adhesion of peripheral blood lymphocytes (PBL) to activated human umbilical vein endothelial cells (HUVEC) under conditions that resemble blood flow. METHODS: Assays of adhesion of PBL to HUVEC or recombinant vascular cell adhesion molecule 1 (rVCAM-1), intercellular adhesion molecule 1 (ICAM-1), and E-selectin were performed under continuous rotation at 37 degrees C. The phenotype of PBL subpopulations attached was characterized by flow cytometry. Lymphocytes were pretreated with different doses (5-100 microg/ml) of aceclofenac, diclofenac, indomethacin, or piroxicam or with inhibitory monoclonal antibodies (MAb) prior to the adhesion assays. The effect of NSAIDs on lymphocyte adhesion molecules was assessed by flow cytometry. To determine whether NSAIDs interfere with the affinity state of very late activation antigen 4 (VLA-4) integrin, we studied the effect of these drugs on the appearance of a beta1 activation-dependent epitope recognized by the HUTS21 MAb both on human T lymphoblasts and on synovial fluid lymphocytes (SFL). RESULTS: In the flow-resembling model, PBL-HUVEC adhesion was mainly mediated by the VLA-4/ VCAM-1 adhesion pathway. The major PBL subset attached was the CD3+, CD45RO+ memory T cell, with CD49d(high) expression. Aceclofenac, diclofenac, and indomethacin, but not piroxicam, were able to inhibit PBL adhesion to HUVEC or rVCAM-1. However, the quantitative expression of VLA-4 was not affected by treatment of PBL with any of the NSAIDs studied. On T lymphoblasts and SFL, mostly CD45RO+ cells, the expression of the beta1 activation-dependent epitope detected by HUTS21 MAb was significantly decreased by aceclofenac, diclofenac, and indomethacin. CONCLUSION: Some NSAIDs are able to inhibit the adhesion of PBL to HUVEC under conditions that resemble blood flow by interfering with the conformational change in VLA-4 that increases its affinity for VCAM-1. PMID- 9751103 TI - Immunohistochemical and molecular studies of serotonin, substance P, galanin, pituitary adenylyl cyclase-activating polypeptide, and secretoneurin in fibromyalgic muscle tissue. AB - OBJECTIVE: Because former investigations have reported abnormal changes in the expression of serotonin (5-hydroxytryptamine [5-HT]) and substance P (SP) in serum and cerebrospinal fluid, this study sought to determine whether 5-HT and pain-modulating neuropeptides (SP, galanin [GA], pituitary adenylyl cyclase activating polypeptide, and secretoneurin) were expressed abnormally in the muscle tissue of patients with fibromyalgia (FM). METHODS: Snap-frozen muscle tissue specimens (deltoid muscles) from 10 patients with FM (mean disease duration 15 years) and from 10 healthy control subjects were examined by reverse transcriptase-polymerase chain reaction (RT-PCR) of RNA preparations from muscle cells, and by immunohistochemistry methods (alkaline phosphatase-anti-alkaline phosphatase and immunogold-silver) using specific primers as well as antibodies. When specific messenger RNA (mRNA) was detected by RT-PCR, in situ RT-PCR was performed for mRNA localization. RESULTS: Specific mRNA for the examined substances was absent in 9 of 10 FM patients and in 10 of 10 controls. No differences between the FM patients and controls could be detected in the muscle tissue by immunohistochemistry. In 1 FM patient, mRNA for the GA receptor could be shown. CONCLUSION: This study showed that 5-HT and neuropeptides are not produced in the muscle of patients with FM, and therefore do not appear to be involved in the peripheral induction of pain in this chronic, painful disorder. PMID- 9751105 TI - Clinical image: Isolated arteritis of the superior mesenteric artery with positive perinuclear antineutrophil cytoplasmic antibody. PMID- 9751104 TI - Overexpanded B cell clone mediating leukemic arthritis by abundant secretion of interleukin-1beta: a case report. AB - The role of cytokines in leukemic arthritis is unknown. The presentation of a patient with B cell chronic lymphocytic leukemia and destructive arthritis of the wrist joints prompted us to study the synovial cytokine pattern by immunohistologic analysis. In addition, rearranged V(H) and V(L) immunoglobulin genes were sequenced to assess B cell clonality. Heavy infiltrations of CD20+ cells with lambda light chain restriction were found in the synovial tissue. Sequencing demonstrated overexpansion of a single B cell clone (DP58/D/J(H)4b and IGLV3S2/Jlambda2-Jlambda3 for V(H) and V(L), respectively) in the peripheral blood. Identical V(H) and V(L) rearrangements were found in the synovial infiltrates. Somatic mutations were found in both the peripheral blood and the synovial clone. Immunohistologic study revealed the presence of abundant interleukin-1beta (IL-1beta) and, to a lesser degree, tumor necrosis factor beta (TNFbeta) (lymphotoxin). In contrast, TNFalpha, interferon-gamma, IL-4, IL-6, and IL-10 were rarely found in the synovial infiltrates. Therefore, IL-1beta secreted in great amounts by leukemic B cells appears to be the major cytokine that mediates joint destruction in leukemic arthritis. PMID- 9751106 TI - Giant cell arteritis and magnetic resonance angiography. PMID- 9751107 TI - Long-term followup of patients receiving combined therapy with cyclosporine and methotrexate. PMID- 9751108 TI - Paradoxical immunologic effects of 2-CdA therapy: comment on the article by Davis et al. PMID- 9751109 TI - Avocado/soybean unsaponifiables in the treatment of scleroderma: comment on the article by Maheu et al. PMID- 9751110 TI - Questions regarding the design of the study of pulse versus oral cyclophosphamide in the treatment of Wegener's granulomatosis: comment on the article by Guillevin et al. PMID- 9751111 TI - Pulse versus oral cyclophosphamide in the treatment of Wegener's granulomatosis: comment on the article by Guillevin et al. PMID- 9751112 TI - Steroid treatment and osteonecrosis: comment on the article by Yamamoto et al. PMID- 9751113 TI - Preventing Emerging Infectious Diseases: A Strategy for the 21st Century. Overview of the Updated CDC plan. AB - Societal, technological, and environmental factors continue to have a dramatic effect on infectious diseases worldwide, facilitating the emergence of new diseases and the reemergence of old ones, sometimes in drug-resistant forms. Modern demographic and ecologic conditions that favor the spread of infectious diseases include rapid population growth; increasing poverty and urban migration; more frequent movement across international boundaries by tourists, workers, immigrants, and refugees; alterations in the habitats of animals and arthropods that transmit disease; increasing numbers of persons with impaired host defenses; and changes in the way that food is processed and distributed. Several recent health events underscore the need for a public health system ready to address whatever disease problems that might arise. For example, in 1997, an avian strain of influenza that had never before infected humans began to kill previously healthy persons in Hong Kong, and strains of Sta phylococcus aureus with diminished susceptibility to the antibiotic vancomycin were reported in Japan and the United States. In addition, researchers recently discovered that a strain of the virus that causes acquired immunodeficiency syndrome (AIDS) had been infecting humans for at least 20 years before AIDS emerged as a worldwide epidemic. Preventing Emerging Infectious Diseases: A Strategy for the 21st Century describes CDC's plan to combat today's infectious diseases and prevent those of tomorrow. It represents the second phase of the effort launched in 1994 with the publication of CDC's Addressing Emerging Infectious Disease Threats: A Prevention Strategy for the United States. This overview of the updated plan outlines specific objectives under four major goals: a) surveillance and response, b) applied research, c) infrastructure and training, and d) prevention and control. Achieving these objectives will enhance understanding of infectious diseases and bolster their detection, control, and prevention. The plan also targets nine categories of problems that cause human suffering and place a burden on society. The aim of this plan is to build a stronger, more flexible U.S. public health system that is well-prepared to respond to known disease problems, as well as to address the unexpected, whether it be an influenza pandemic, a disease caused by an unknown organism, or a bioterrorist attack. The implementation of this plan will require the dedicated efforts of many partners, including state and local health departments, other federal agencies, professional societies, universities, research institutes, health-care providers and organizations, the World Health Organization, and many other domestic and international organizations and groups. PMID- 9751114 TI - Mutation of Thr115 in MyoD positively regulates function in murine fibroblasts and human rhabdomyosarcoma cells. AB - Committed skeletal muscle myoblasts undergo terminal differentiation when shifted from a high-mitogen medium to a low-mitogen medium. However, expression of the myogenic regulatory factor MyoD seems to be similar in proliferating and differentiating cells, suggesting that its function is attenuated in proliferating myoblasts. To further understand the potential mechanisms that may attenuate MyoD function, we have examined the effect of posttranslational modification. By analogy with myogenin, we have examined the role of phosphorylation in regulating the function of MyoD. MyoD contains two putative protein kinase C (PKC) phosphorylation sites (Thr115 and Ser200). The former site is analogous to Thr85 within the highly conserved basic domain of myogenin that has been demonstrated to negatively regulate the myogenic differentiation functions of myogenin. To test whether hyperphosphorylation of the same PKC site in MyoD would attenuate its function, we generated a mutant MyoD with a single amino acid substitution (Thr115-Ala) that disrupts the PKC phosphorylation site (Thr115) within the conserved basic domain. Wild-type and mutant MyoD were introduced into cells using an E1, E3-deleted adenoviral vector. In mouse C3H10T1/2 fibroblasts, both wild-type and mutant MyoD induced terminal myogenic differentiation when growth factors were withdrawn from the cell culture. Consistent with these results, nuclear extracts from infected cells, but not those from uninfected cells, demonstrated complex formation with an oligonucleotide containing an E-box consensus sequence. Growth arrest was associated with the up-regulation of p21cip1, cell fusion to multinucleated myotubes, and the expression of a muscle differentiation marker (myosin heavy chain). On the other hand, when infected cells were maintained under high mitogenic conditions (in the presence of 10% fetal bovine serum), the expression of wild-type or mutant MyoD slowed cell growth and induced p21cip1. Only mutant MyoD caused cell fusion, myosin heavy chain expression, and altered mobility of the E-box oligonucleotide in gel shift assays. Furthermore, after infection, MyoD was phosphorylated, and phosphothreonine was detected in wild-type MyoD immunoprecipitated only from C3H10T1/2 cells grown under high mitogenic conditions. These results suggest that Thr115 may play an important role in the regulation of MyoD function under conditions of high mitogenesis. MyoD was also phosphorylated in malignant rhabdomyosarcoma (RMS) cells in which MyoD function was attenuated. Phosphothreonine was also detected in MyoD immunoprecipitates. Rh30 alveolar RMS cells were infected with an adenovirus expressing either wild type or mutant MyoD. In contrast to the results in fibroblasts, when overexpressed in malignant Rh30 RMS cells, mutant MyoD arrested cell growth without inducing p21cip1 and caused cell fusion. However, no muscle differentiation markers were detected, indicating that an overexpression of mutant MyoD lacking Thr115 caused Rh30 cells to become quiescent and recapitulate at least some aspects of myogenesis (cell fusion). PMID- 9751115 TI - Deregulated expression of the retinoid X receptor alpha prevents muscle differentiation in P19 embryonal carcinoma cells. AB - We have studied the expression of the retinoid X receptor (RXR) family of receptors during the DMSO-induced differentiation of P19 murine embryonal carcinoma cells into mesoderm and muscle. RXR-alpha protein is weakly detectable in untreated P19 cells and in a mutant line of P19 cells (D3) that are resistant to DMSO-induced differentiation but begins to increase by day 3 and continues to rise gradually thereafter, whereas RXR-gamma protein is readily detected in P19 cells and decreases over the course of differentiation. Protein expression is uncoupled from mRNA levels, because DMSO induces a rapid, aggregation independent, transient increase in RXR-alpha mRNA that diminishes by day 3 of differentiation. Thus, the expression of RXR-alpha protein is prevented at early times during DMSO-induced differentiation. Stable P19 cell clones that constitutively express RXR-alpha protein [P19(RXR-alpha)] are resistant to DMSO induced differentiation associated with increased levels of oligonucleosomal length DNA fragmentation. Loss of RXR-alpha expression after multiple passages results in a reversion to a DMSO-responsive phenotype. Id1 transcripts are present in P19 cells and are transiently decreased on day 2 of DMSO differentiation but remain elevated in DMSO-treated P19(RXR-alpha) and in P19 cells treated simultaneously with retinoic acid and DMSO. The mRNA for the mesoderm inducer protein Brachyury T was also deregulated in P19(RXR-alpha) cells and D3 cells compared with that of wild-type P19 cells. Together, these results show that expression of the RXR-alpha mRNA and protein in P19 cells is tightly regulated during the mesodermal/muscle differentiation of P19 cells, and that ectopic expression of the RXR-alpha protein prevents differentiation associated with increased cell death, prolonged expression of Brachyury T, and constitutive expression of Id1. PMID- 9751116 TI - Basic fibroblast growth factor-induced decrease in type I collagen gene transcription is mediated by B-myb. AB - Basic fibroblast growth factor (bFGF), a member of the fibroblast growth factor family, potently induces increased vascular smooth muscle cell (SMC) proliferation and decreased expression of type I collagen. Recently, our laboratory demonstrated that, in bovine vascular SMCs, expression of B-myb, a member of the myb gene family, is dependent upon cellular growth state and that B myb decreases alpha1(I) collagen promoter activity in transient transfection assays. Nuclear run-off analysis indicated that the decrease in alpha1(I) collagen mRNA level seen upon bFGF treatment was due to a decline in the rate of alpha1(I) procollagen gene transcription. Thus, we investigated the potential role of B-Myb in the down-regulation of type I collagen gene expression by bFGF. Using Northern blot analysis, we found that bFGF treatment of bovine aortic SMCs caused an increase in B-myb mRNA levels. Ectopic expression of B-myb decreased endogenous alpha1(I) collagen mRNA levels. Importantly, introduction of a B-myb antisense oligonucleotide prevented the drop in the alpha1(I) collagen mRNA levels seen upon treatment with bFGF. Together, these results indicate that B-myb mediates signals leading to the decreased rate of alpha1(I) collagen gene transcription caused by bFGF. PMID- 9751117 TI - Differential apoptotic behaviors of c-myc, N-myc, and L-myc oncoproteins. AB - c-, N-, and L-myc are related nuclear oncoproteins that bind similar DNA sites and cooperate with activated ras oncogenes to transform primary fibroblasts. Although c-myc can also promote apoptosis in some cells after growth factor withdrawal or exposure to cytotoxic agents, roles for N- and L-myc in apoptosis remain undetermined. To address this, c-, N-, or L-myc were stably expressed in the interleukin 3 (IL-3)-dependent 32D hematopoietic cell line. The apoptotic response of each cell line was assessed after IL-3 withdrawal or treatment with four structurally unrelated cytotoxic agents. All three oncoproteins accelerated apoptosis after IL-3 withdrawal. In contrast, whereas c-myc overexpression generally sensitized cells to cytotoxic drugs, N-myc and L-myc overexpression produced resistance. myc expression tended to be associated with a more robust G2 M arrest after drug exposure, but this did not correlate with drug sensitivity or resistance. Bcl-2 and Bcl-X(L) protected control cells against apoptosis after either IL-3 withdrawal or drug exposure, although in some cases this effect could be overridden by myc oncoproteins, particularly N-myc and L-myc. Our results suggest that the apoptotic pathways activated upon IL-3 withdrawal and cytotoxic drug treatment are distinct and differentially affected by members of the myc and Bcl-2 families. PMID- 9751118 TI - Nuclear redistribution of BRCA1 during viral infection. AB - The functions and the intracellular localization of the breast/ovarian susceptibility gene product, BRCA1, has been controversial. To arrive at a clear understanding of its localization and relative position to other nuclear structures, a new monoclonal antibody was produced and characterized by immunohistochemical techniques with other BRCA1 antibodies. Each of the antibodies specifically detected BRCA1 as localized to specific nuclear domains and did so in a variety of cells and in a cell cycle-dependent manner. However, all antibodies also cross-reacted with the centrosomal domain, suggesting that BRCA1 is also localized to this important mitotic component. We found that the BRCA1-containing nuclear domains are different than any of the well-defined nuclear domains. However, a cell cycle-related partial overlap was found for HP1alpha, a chromo-domain-containing protein involved in heterochromatin maintenance. Cellular stimuli, such as heat shock and herpes virus infection, dispersed BRCA1 from its domains. In contrast, infection with adenovirus 5 recruited BRCA1 to regions of viral transcription and replication. These disparate distributions of BRCA1 may provide clues to its function. PMID- 9751119 TI - The Src homology 2 domain protein Shb transmits basic fibroblast growth factor- and nerve growth factor-dependent differentiation signals in PC12 cells. AB - To assess a possible role for the Src homology 2 (SH2) domain adaptor protein Shb in PC12 cell signal transduction and differentiation, we have investigated the expression of Shb in PC12 cells and found that the differentiation factors nerve growth factor (NGF) and basic fibroblast growth factor (bFGF), as well as the PC12 cell mitogen epidermal growth factor, increased Shb protein expression and Shb mRNA steady-state levels. Two PC12 cell clones stably overexpressing the Shb cDNA exhibited enhanced NGF- or bFGF-induced differentiation, assessed as neurite outgrowth. This effect showed no direct correlation to mitogen-activated protein kinase activation, although the mitogen-activated protein kinase/kinase inhibitor PD-98059 caused a partial inhibition of neurite outgrowth. Furthermore, it was found that the Shb-overexpressing cells extended neurites in response to epidermal growth factor. The effects of Shb overexpression on neurite outgrowth required a functional SH2 domain because PC12 cells expressing Shb with an inactivated SH2 domain did not differentiate more readily in response to NGF. Tyrosine phosphorylation of the p13 Suc1-associated neurotrophic factor-induced tyrosine-phosphorylated target protein in response to bFGF was strongly inhibited by Shb overexpression, without correlating with the corresponding rate of neurite outgrowth. NGF-induced tyrosine phosphorylation of phospholipase Cgamma, TrkA, and Shc was unaltered in the Shb-overexpressing cells, whereas that of Shb was greatly enhanced in these cells compared with control PC12-neo cells. In addition, an NGF-activated Mr 140,000 phosphotyrosine protein was found to be associated with Shb in immunoprecipitation experiments. The data in this study suggest that Shb is involved in transmission of NGF- and bFGF-dependent differentiation signals in PC12 cells. PMID- 9751120 TI - Role of mitogen-activated protein kinase in taxol-induced apoptosis in human leukemic U937 cells. AB - The induction of apoptosis by Taxol was investigated in human leukemic U937 cells. Treatment of U937 cells with 20 nM Taxol for 24 h induced apoptosis in 30 40% of cells, which resulted in an 80% growth inhibition 3 days after treatment. Synchronous cells at different cell cycle stages exhibited different sensitivities toward Taxol, and their reversion by certain protein kinase inhibitors was also phase specific. Kinetic studies of cell cycle progress reveal that Taxol accelerates the progression of the cell cycle, which facilitates the process of apoptosis, especially for cells initially in the G1 phase. This acceleration may result from transient activation of p42/ 44 mitogen-activated protein (MAP) kinase, because inhibition of upstream MAP/extracellular signal regulated kinase kinase (MEK1/2) by PD98059 reversed this effect. However, the delayed S-G2-M-phase progression by PD98059 was insignificant. The results suggest that MAP kinase may not only mediate cell cycle progress but may also participate in the apoptosis pathway for cells originally in S phase. PMID- 9751122 TI - The absence of p27Kip1, an inhibitor of G1 cyclin-dependent kinases, uncouples differentiation and growth arrest during the granulosa->luteal transition. AB - The involvement of cyclin-dependent kinase inhibitors in differentiation remains unclear: are the roles of cyclin-dependent kinase inhibitors restricted to cell cycle arrest; or also required for completion of the differentiation program; or both? Here, we report that differentiation of luteal cells can be uncoupled from growth arrest in p27-deficient mice. In these mice, female-specific infertility correlates with a failure of embryos to implant at embryonic day 4.5. We show by ovarian transplant and hormone reconstitution experiments that failure to regulate luteal cell estradiol is one physiological mechanism for infertility in these mice. This failure is not due to a failure of p27-deficient granulosa cells to differentiate after hormonal stimulation; P450scc, a marker for luteal progesterone biosynthesis, is expressed and granulosa cell-specific cyclin D2 expression is reduced. However, unlike their wild-type counterparts, p27 deficient luteal cells continue to proliferate for up to 3.5 days after hormonal stimulation. By day 5.5, however, these cells withdraw from the cell cycle, suggesting that p27 plays a role in the early events regulating withdrawal of cells from the cell cycle. We have further shown that in the absence of this timely withdrawal, estradiol regulation is perturbed, explaining in part how fertility is compromised at the level of implantation. These data support the interpretation of our previous observations on oligodendrocyte differentiation about a role for p27 in establishing the nonproliferative state, which in some cases (oligodendrocytes) is required for differentiation, whereas in other cases it is required for the proper functioning of a differentiated cell (luteal cell). PMID- 9751121 TI - Activation and function of the epidermal growth factor receptor and erbB-2 during mammary gland morphogenesis. AB - The hormonal stimulation of mammary gland morphogenesis is believed to occur through growth factor receptor signaling pathways. To determine the importance of the epidermal growth factor receptor (EGFR) pathway, we examined extracts of inguinal mammary glands from prepubertal and pubertal mice for tyrosine phosphorylated EGFR and other erbB receptors. Tyrosine phosphorylation of both EGFR and erbB-2 was detected in normal female BALB/c mice at 5-6 weeks of age, but not during the prepubertal stage, e.g., 24 days of age. Treatment of mice with estradiol or epidermal growth factor also stimulated the formation of mammary EGFR/erbB-2 phosphotyrosine. Waved-2 mice, which have impaired EGFR kinase activity, exhibited less mammary development than did wild-type (wt) mice when both were evaluated at 36 days of age. Because EGFR knockout (KO) mice die shortly after birth, glands from the newborns were implanted under the renal capsules of female nude mice. Under these conditions, extensive ductal growth was observed in mammary glands from wt animals; in contrast, glands from EGFR KO mice failed to grow beyond rudimentary structures. Tissue recombinants revealed that the wt fat pad supported the morphogenesis of EGFR KO epithelium, whereas the EGFR KO fat pad did not. Taken together, these data suggest that EGFR is essential for morphogenesis of the mammary ducts and functions during this period of mammary development as a heterodimer with erbB-2 in the mammary stroma. PMID- 9751123 TI - Functional rescue of Stat5a-null mammary tissue through the activation of compensating signals including Stat5b. AB - Prolactin induces mammopoiesis and lactogenesis through the Janus kinase-signal transducers and activators of transcription pathway, with Stat5a being a principal and obligate cytoplasmic and nuclear signaling molecule. Mice from which the Stat5a gene has been deleted fail to develop functional mammary tissue during their first pregnancy. Lobuloalveolar outgrowth is curtailed, and epithelial cells fail to progress to functional differentiation. Here, we investigate whether the effect of Stat5a deficiency is restricted to the epithelium and whether the gland has the capacity to activate alternative signaling pathways that could restore development and function. Mammary gland transplant experiments showed that Stat5a-deficient epithelium does not differentiate in wild-type stroma, thus demonstrating a cell-autonomous role for Stat5a. The capacity of Stat5a-deficient mammary tissue to develop and secrete milk was measured after consecutive pregnancies and with postpartum suckling. Neither of these regimens could independently restore lactation. However, the combination of several pregnancies and suckling stimuli resulted in a partial establishment of lactation and an increase of Stat5b activity. These experiments demonstrate that the mammary gland has inherent plasticity that allows it to use different signals to achieve its ultimate purpose, the production of milk to nurture newborn offspring. PMID- 9751124 TI - Expression of the RNA component of telomerase during human development and differentiation. AB - We used a radioactive in situ method to study expression of the RNA component of human telomerase (hTR) during normal human development and differentiation using archival tissues. In embryonic tissues, the highest and most uniform expression was present in undifferentiated neuroepithelium. Expression was stronger in immature epithelium than in accompanying immature mesenchyme. Differentiation of most tissues was accompanied by decreased or absent expression. Except for testis and adrenal, the adult pattern of expression was present by the 10th postnatal week. In adult tissues, high expression was present in the testis (primary spermatocytes and Sertoli cells), moderate expression was present in lymphoid follicles (germinal centers), and weak expression was present in epithelia (regenerative cells) but was absent in the nervous system and mesenchymal derived tissues. Expression in adult tissues was predominantly limited to dividing cells, although certain differentiated postmitotic cells expressed the hTR. Our studies demonstrate the complex interrelationship of hTR expression with human development, differentiation, and cell division. PMID- 9751126 TI - Characterization of a directional selective inhibitory input from the medial terminal nucleus to the pretectal nuclear complex in the rat. AB - The receptive field properties of neurons in the medial terminal nucleus of the accessory optic system (MTN) that project to the ipsilateral nucleus of the optic tract (NOT) and dorsal terminal nucleus (DTN), as identified by antidromic electrical activation, were analysed in the anaesthetized rat. The great majority (88%) of MTN neurons that were antidromically activated from NOT and DTN preferred downward directed movement of large visual stimuli while the remaining cells preferred upward directed stimulus movement. Distinct retrograde tracer injections into the NOT/DTN and the ipsilateral inferior olive (IO) revealed that no MTN neurons project to both targets. MTN neurons projecting to the ipsilateral NOT/DTN were predominantly found in the ventral part of the MTN, whereas those projecting to the IO were found in the dorsal part of the MTN. In situ hybridization for glutamic acid decarboxylase (GAD) mRNA was used as a marker for GABAergic neurons. Up to 98% of MTN neurons retrogradely labelled from the ipsilateral NOT/DTN also expressed GAD mRNA. Earlier studies have shown that MTN neurons that prefer upward directed stimulus movements are segregated from MTN neurons that prefer downward directed stimulus movements. It also has been demonstrated that directionally selective neurons in the NOT/DTN prefer horizontal stimulus movements and receive an inhibitory input from ipsilateral MTN. Our results indicate that this input is mediated by GABAergic cells in the ventral part of MTN, which to a large extent prefer downward directed stimulus movements, and that the great majority of MTN neurons that prefer upward directed stimulus movements project to other targets one of which possibly is the IO. PMID- 9751125 TI - Regulation of apoptosis in mouse hepatocytes and alteration of apoptosis by nongenotoxic carcinogens. AB - Regulation of apoptosis is an important component of multistage hepatocarcinogenesis. The objectives of the present study were to characterize apoptosis regulation in primary mouse hepatocytes and to determine whether nongenotoxic carcinogens alter apoptosis regulation. Bleomycin-induced apoptosis was accompanied by decreases in bcl-2 and bcl-xl and increases in p53, bak, and bax protein levels. Transforming growth factor (TGF)-beta-induced apoptosis was accompanied by decreased bcl-xL and increased bak. Bleomycin-induced apoptosis was partially dependent on p53, whereas TGF-beta-induced apoptosis was independent of p53. Phenobarbital inhibited both TGF-beta and bleomycin-induced apoptosis and the normal regulation of p53, bcl-2, and bax. Nafenopin inhibited apoptosis through a mechanism dependent on PPAR-alpha and inhibited the normal regulation of bcl-2 and bak. 2,3,7,8-Tetrachlorodibenzo-p-dioxin did not alter apoptosis or its regulation. Apoptosis was increased in hepatocytes from bcl-2 null mice, which indicated that the bcl-2 family contributes to hepatocyte apoptosis regulation. This study demonstrated that apoptosis regulation in mouse hepatocytes involves distinct pathways and that diverse nongenotoxic carcinogens differentially alter molecular pathways that represent targets for hepatocarcinogenesis. PMID- 9751127 TI - UV-sensitive input to horizontal cells in the turtle retina. AB - Microspectrophotometry, electroretinography and behavioural studies have indicated that ultraviolet (UV) light contributes to functional vision in various vertebrate species. Based on behavioural evidence, this was also suggested for turtle vision. In order to reveal the interactions underlying detection of UV light in the distal retina, we recorded intracellularly the photoresponses of cones and horizontal cells in retinas of Pseudemys scripta elegans and Mauremys caspica and calculated the action spectra of these cells under different conditions of adaptation. In the dark-adapted retina, all three types of horizontal cells; luminosity-type, red/green chromaticity-type and yellow/blue chromaticity-type exhibited increased sensitivity in the UV region of the spectrum. However, chromatic adaptation indicated that only the yellow/blue chromaticity-type horizontal cells received excitatory input from UV-sensitive cones with peak sensitivity approximately 360 nm. The enhanced UV sensitivity of luminosity-type horizontal cells probably reflected the beta-band of the long wavelength sensitive visual pigment as indicated by the action spectra of dark adapted L-cones. It is suggested that the enhanced UV sensitivity of red/green chromaticity-type horizontal cells reflects the beta-band of the medium wavelength sensitive visual pigment. Transmission measurements of the optical media (cornea, lens and vitreous) indicated that UV vision can be functional under normal circumstances. PMID- 9751128 TI - The NMDA receptor subunit NR2B of neonatal rat brain: complex formation and enrichment in axonal growth cones. AB - The N-methyl-D-aspartate (NMDA) subtype of ionotropic glutamate receptors comprises a family of highly homologous subunits which assemble into oligomeric protein complexes. Alterations in subunit composition are developmentally regulated, leading to functionally distinct receptor populations. Here, the contribution of the subunit NR2B to NMDA receptor complex formation was analysed in neonatal rat brain, employing polyclonal antibodies raised against NR2B specific synthetic peptides. By hydrodynamic size fractionation of the solubilized receptor protein and chemical cross-linking, NR2B antigen was found to be associated with several protein species of up to 690 kDa molecular weight. These observations show NR2B to be part of a multimeric receptor complex. Fractionation of cortex homogenates from E18 rat embryos on sucrose density gradients revealed NR2B polypeptide to be highly enriched in axonal growth cones. A similar distribution was found by fluorescence microscopy of immature hippocampal neurons, showing a preferential accumulation of NR2B antigen in axonal growth cones and varicosities. In mature cells, NR2B antigen displayed a punctated distribution pattern with redistribution to somato-dendritic spheres. The association of NR2B with axonal growth cones and processes of immature neurons suggests a role of NMDA receptors in the regulation of neurite outgrowth and migration. PMID- 9751129 TI - The constructive nature of vision: direct evidence from functional magnetic resonance imaging studies of apparent motion and motion imagery. AB - Echoplanar functional magnetic resonance imaging was used to monitor activation changes of brain areas while subjects viewed apparent motion stimuli and while they were engaged in motion imagery. Human cortical areas MT (V5) and MST were the first areas of the 'dorsal' processing stream which responded with a clear increase in signal intensity to apparent motion stimuli as compared with flickering control conditions. Apparent motion of figures defined by illusory contours evoked greater activation in V2 and MT/MST than appropriate control conditions. Several areas of the dorsal pathway (V3A, MT/MST, areas in the inferior and superior parietal lobule) as well as prefrontal areas including FEF and BA 9/46 responded strongly when subjects merely imagined moving stimuli which they had seen several seconds before. The activation during motion imagery increased with the synaptic distance of an area from V1 along the dorsal processing stream. Area MT/MST was selectively activated during motion imagery but not during a static imagery control condition. The comparison between the results obtained with objective motion, apparent motion and imagined motion provides further insights into a complex cortical network of motion-sensitive areas driven by bottom-up and top-down neural processes. PMID- 9751130 TI - Eph receptor-ligand interactions are necessary for guidance of retinal ganglion cell axons in vitro. AB - Previous results of an in vitro guidance test, the stripe assay, have demonstrated the presence of a repulsive axon guidance activity for temporal retinal axons in the posterior part of the vertebrate optic tectum. Ephrin-A5 and Ephrin-A2 are ligands for the EphA subfamily of Eph receptor tyrosine kinases, which are expressed in overlapping gradients in the posterior part of the tectum. When recombinantly expressed, both proteins have been shown to guide retinal ganglion cell axons in the stripe assay. While these results suggest that Ephrin A5 and Ephrin-A2 form part of the posterior repulsive guidance activity, they do not elucidate whether they are necessary components. Here we report that soluble forms of the ligands at nanomolar concentrations completely abolish this repulsive activity. Similar results were obtained with the soluble extracellular domain of EphA3, which is a receptor for Ephrin-A2 and Ephrin-A5, but not with the corresponding domain of EphB3, a receptor for the transmembrane class of Eph ligands. These experiments show that the repulsive axon guidance activity seen in the stripe assay is mediated by Ephrin-A ligands. PMID- 9751131 TI - Retrograde degeneration of thalamic neurons in the mediodorsal nucleus after neonatal and adult aspiration lesions of the medial prefrontal cortex in the rat. Implications for mechanisms of functional recovery. AB - The behavioural consequences of neonatal lesions of the frontal cortex are limited as compared with similar lesions performed in adulthood. The present study has investigated, using unbiased quantitative methods with randomized systematic sampling, the total neuronal cell numbers in the mediodorsal nucleus of the thalamus after aspiration lesions of the medial prefrontal cortex performed in neonatal and in adult rats. It was found that the reduction in total cell numbers after neonatal prefrontal cortex lesions was similar to that found after adult cortex lesions. In neonatally lesioned animals the neuronal cell density was significantly increased by 13%, whereas in adult lesioned animals it was unchanged. On the other hand, the volume of the mediodorsal nucleus was reduced by 27% in neonatally, and 20% in adult lesioned animals. Total neuronal cell number of the mediodorsal nucleus was significantly decreased in neonatally as well as in adult lesioned rats, by 14% and 21%, respectively. These findings are discussed in the light of the previously proposed role of the thalamus as a neural substrate of functional sparing after neonatal cortical lesions. PMID- 9751133 TI - Locomotor-like movements evoked by leg muscle vibration in humans. AB - We attempted to elicit automatic stepping in healthy humans using appropriate afferent stimulation. It was found that continuous leg muscle vibration produced rhythmic locomotor-like stepping movements of the suspended leg, persisting up to the end of stimulation and sometimes outlasting it by a few cycles. Air-stepping elicited by vibration did not differ from the intentional stepping under the same conditions, and involved movements in hip and knee joints with reciprocal electromyogram (EMG) bursts in corresponding flexor and extensor muscles. The phase shift between evoked hip and knee movements could be positive or negative, corresponding to 'backward' or 'forward' locomotion. Such an essential feature of natural human locomotion as alternating movements of two legs, was also present in vibratory-evoked leg movements under appropriate conditions. It is suggested that vibration evokes locomotor-like movements because vibratory-induced afferent input sets into active state the central structures responsible for stepping generation. PMID- 9751132 TI - Changes in the expression of protease-activated receptor 1 and protease nexin-1 mRNA during rat nervous system development and after nerve lesion. AB - HDs racI Thrombin causes profound metabolic and morphological changes in cultured neural cells via activation of the thrombin receptor, also called protease activated receptor 1 (PAR1). PAR1 mRNA is present in the rat brain, but the role of this receptor in the nervous system remains elusive. The expression of PAR1 and the potent thrombin inhibitor protease nexin-1 (PN-1) was investigated in the developing rat brain and spinal cord and after peripheral nerve lesion. As seen by in situ hybridization, the PAR1 mRNA signal in the late embryonic and early postnatal nervous system was widespread, but generally of low intensity whereas in the adult it was more pronounced and confined to particular neuronal cells. These include the mesencephalic dopaminergic neurons, several thalamic and brainstem nuclei, the mitral cells in the olfactory bulb and the Purkinje cells in the cerebellum. In the spinal cord, PAR1 mRNA was abundant in motoneurons and a particularly high expression was detected in the preganglionic neurons of the autonomic nervous system. High PAR1 mRNA expression was also found in the dorsal root ganglia. Interestingly, strong immunoreactivity for the protease inhibitor PN-1 was present in spinal motoneuron cell bodies, although its transcript was undetectable there. In response to sciatic nerve transection, the signal intensity of PAR1 mRNA as seen by Northern analysis increased in the proximal and the distal part of the lesioned nerve and in the denervated muscle, whereas the PN-1 mRNA signal strongly increased only in the distal part of the nerve but remained unchanged in the proximal part and in the muscle. After facial nerve transection, PAR1 mRNA expression substantially decreased in facial motoneurons. No PAR1 transcript was detected in reactive astrocytes. Similar to PAR1, PN-1 mRNA which was expressed in interneurons within the facial nucleus was also decreased following facial nerve transection. PMID- 9751134 TI - Microglial NO induces delayed neuronal death following acute injury in the striatum. AB - We have established a novel injury model in the central nervous system by a stereotaxic injection of ethanol into rat striatum to induce necrosis. With this model, we clarify a function of inducible nitric oxide synthase (iNOS) in a healing mechanism around a necrotic lesion. A semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR) revealed that the iNOS mRNA arose at 6 h, peaked at 24 h, and declined to a lower level 48 h after an intrastriatal 5-microL ethanol injection. From in situ hybridization, this iNOS mRNA was expressed in the area surrounding the injury. By immunohistochemistry, mononuclear cells at this boundary area of necrosis were stained with anti-iNOS antibody on the first day after the injury. These cells turned out to be reactive microglia from the positive staining of GSA-I-B4, ED-1 and OX-42. Haematoxylin eosin (HE) staining showed that neurons in this boundary area gradually disappear up to 5 days after the injury with an increment of microglial cells, and this area became cavernous. Nuclei of neurons in this area were stained positive by the terminal deoxynucleotidyl-transferase-mediated dUTP-biotin nick end-labelling (TUNEL) assay on the first day after the injury. These TUNEL-positive neurons gradually disappeared toward the third day, while microglial cells increased. L Ng-nitro-arginine methylester (L-NAME), a competitive NOS inhibitor, administration diminished the elimination of neurons by microglia in this boundary area surrounding necrosis. Microglial NO may act as a neurotoxic agent to eliminate damaged neurons near the necrosis in the form of delayed neuronal death, and may reintegrate the neuronal circuits with functionally intact neurons. PMID- 9751135 TI - Transcripts for secreted and GPI-anchored brevican are differentially distributed in rat brain. AB - Brevican is a member of the aggrecan/versican family of proteoglycans. In contrast to the other family members, brevican occurs both as soluble isoforms secreted into the extracellular space and membrane-bound isoforms which are anchored to the cell surface via a glycosylphosphatidylinositol (GPI) moiety. Expression of both variants, which are encoded by two differentially processed transcripts from the same gene, is confined to the nervous system. In the current study, we have used in situ hybridization to examine the cellular sites of synthesis for both mRNAs during postnatal development of the rat brain. Whereas the 3.6-kb transcript encoding secreted brevican displays a widespread distribution in grey matter structures, including cerebellar and cerebral cortex, hippocampus and thalamic nuclei with silver grains accumulating over neuronal cell bodies, the smaller transcript (3.3 kb) encoding GPI-anchored isoforms appears to be largely confined to white matter tracts and diffusely distributed glial cells. This expression pattern is further confirmed by reverse transcriptase-polymerase chain reaction (RT-PCR) experiments with RNA from different glial cell cultures, and by biochemical data demonstrating that the crude membrane fraction from isolated optic nerve contains high amounts of phosphatidylinositol-specific phospholipase C (PI-PLC)-sensitive brevican immunoreactivity. During ontogenetic development, both brevican transcripts are generally up-regulated. However, the expression of glypiated brevican is delayed by about 1 week, compared with the expression of the secreted isoform. This late appearance of GPI-linked brevican, its predominant expression in glial cells and its tight association with brain myelin fractions suggest a functional role in neuroglia. PMID- 9751136 TI - Accumulation of acetylcholine receptors is a necessary condition for normal accumulation of acetylcholinesterase during in vitro neuromuscular synaptogenesis. AB - To study a step of the very complex processes of the formation of the neuromuscular junction (NMJ), we have analysed the clustering of acetylcholine receptors (AChR) and acetylcholinesterase (AChE) in myotubes cultured in various conditions. On the surface of rat myotubes cultured in the presence of spinal cord cells from embryonic rat, numerous AChE clusters appeared. Such clusters are always co-localized with AChR clusters, but the reverse is not true: the number of AChR clusters largely exceeds that of AChE clusters. Very few AChE clusters formed when such co-cultures were treated with monoclonal antibodies (mAbs) against the main immunogenic region (MIR) of the AChR, which provoke internalization and degradation of the AChRs of the muscular membrane. The total levels of AChE and proportions of molecular forms were unaffected. We also used non-innervated myotubes in which addition of agrin, a protein normally synthesized by motoneurons, transported to nerve terminals and inserted into the synaptic basal lamina, induces the formation of small clusters of AChE. When added to rat myotubes devoid of membrane AChR, agrin-induced AChE clusters did not form. Finally, we analysed the capacity of the variant of the C2 mouse muscle cell line deficient in AChR (1R-) to form clusters of AChE in co-cultures with spinal cord cells from rat: no formation of AChE clusters could be observed. In all these different systems of cultures, the conditions which prevented clustering of AChR (anti-AChR antibodies, deficiency of the variant C2 cell line) also suppressed AChE clustering. We concluded that clustering of AChR is a prerequisite for clustering of AChE, so that NMJ formation implies the sequential accumulation of these two components. PMID- 9751137 TI - Study of subcellular localization of membrane-bound choline acetyltransferase in Drosophila central nervous system and its association with membranes. AB - Choline acetyltransferase (ChAT), the enzyme which catalyses the biosynthesis of the neurotransmitter acetylcholine, exists in a soluble and membrane-bound form in cholinergic nerve terminals of different animal species. This study was performed on the enzyme present in Drosophila central nervous system. We show that the two forms of the enzyme have the same apparent molecular weight (75 kDa) when analysed by immunoblotting using an antibody we raised against the recombinant enzyme. According to different authors, membrane-bound enzyme might be associated with synaptic vesicles or plasma membrane. Subfractionation of Drosophila head homogenates in linear glycerol gradients showed that ChAT does not associate with synaptic vesicles. Analysis of ChAT activity and immunoreactivity showed that two peaks of ChAT were produced. One peak was present in fractions containing soluble components and the other was associated with rapidly sedimenting membranes containing plasma membranes. ChAT in the first peak was mainly hydrophilic. A large proportion of ChAT associated with rapidly sedimenting membranes was amphiphilic. Further fractionation of these membranes by flotation in sucrose gradients showed that membrane-associated ChAT sedimented in fractions containing plasma membrane marker. Membrane-bound ChAT was neither solubilized nor converted to hydrophilic enzyme after membrane treatment with 1 M hydroxylamine, suggesting that the enzyme is not palmitoylated and therefore not anchored to membrane through thioester-linked long chain fatty acid. Partial solubilization of ChAT present on membranes with urea and carbonate suggests that this form of ChAT is a peripheral membrane protein. Carbonate solubilization of membrane-bound ChAT converted the enzyme from hydrophobic to hydrophilic protein. PMID- 9751138 TI - G-proteins and G-protein subunits mediating cholinergic inhibition of N-type calcium currents in sympathetic neurons. AB - One postsynaptic action of the transmitter acetylcholine in sympathetic ganglia is to inhibit somatic N-type Ca2+ currents: this reduces Ca2+-activated K+ currents and facilitates high-frequency spiking. Previous experiments on rat superior cervical ganglion neurons have revealed two distinct pathways for this inhibitory action: a rapid, voltage-dependent inhibition through activation of M4 muscarinic acetylcholine receptors (mAChRs), and a slower, voltage-independent inhibition via M1 mAChRs [Hille (1994) Trends in Neurosci., 17, 531-536]. We have analysed the mechanistic basis for this divergence at the level of the individual G-proteins and their alpha and betagamma subunits, using a combination of site directed antibody injection, plasmid-driven antisense RNA expression, overexpression of selected constitutively active subunits, and antagonism of endogenously liberated betagamma subunits by over-expression of Dy-binding P adrenergic receptor kinase 1 (PARK1) peptide. The results indicate that: (i) M4 mAChR-induced inhibition is mediated by GoA; (ii) a and Py subunits released from the activated GoA heterotrimer produce separate voltage-insensitive and voltage sensitive components of inhibition, respectively; and (iii) voltage-insensitive M1 mAChR-induced inhibition is likely to be mediated by the alpha subunit of Gq. Hence, Ca2+ current inhibition results from the concerted, but independent actions of three different G-protein subunits. PMID- 9751139 TI - Differential regulation of FAK+ and PYK2/Cakbeta, two related tyrosine kinases, in rat hippocampal slices: effects of LPA, carbachol, depolarization and hyperosmolarity. AB - FAK+, an isoform of focal adhesion kinase preferentially expressed in brain and PYK2/Cakbeta (proline-rich tyrosine kinase 2/cell adhesion kinasebeta) are two related cytoplasmic tyrosine kinases. They are candidates for coupling electrical activity and stimulation of neurotransmitter receptors to short and long-term changes in synaptic properties, cytoskeletal organization and gene expression in neurons. As the same set of stimuli appear capable of stimulating FAK and/or PYK2 in non-neuronal cells and in cell lines with neuronal characteristics, we investigated the selectivity of regulation of these two kinases in mature nervous tissue. Using rat hippocampal slices, we compared the regulation of FAK+ and PYK2 by stimuli known to be active on one or the other of these two kinases in other cell types: lysophosphatidic acid (LPA), carbachol, depolarization, and hyperosmolarity. Phosphorylation of FAK+ was markedly increased by carbachol and LPA. Carbachol effects occurred via activation of M1 muscarinic receptors and nicotinic receptors. The effects of carbachol and LPA were prevented by protein kinase C inhibitors, whereas 8-Br-cAMP attenuated the effects of carbachol but not of LPA. Tyrosine phosphorylation of PYK2 but not of FAK+ was very strongly enhanced by depolarization and hyperosmolarity. This study and our previous results show that FAK+ and PYK2 are regulated differentially in hippocampal slices: FAK+ is phosphorylated on tyrosine in response to stimulation of G protein-coupled receptors, whereas PYK2 is mainly sensitive to depolarization and hyperosmolarity. Thus, FAK+ and PYK2 may provide specific and separate links between activation of neurotransmitters receptors, depolarization and tyrosine phosphorylation in mature hippocampus. PMID- 9751140 TI - Coexpression of dopamine D1 and D3 receptors in islands of Calleja and shell of nucleus accumbens of the rat: opposite and synergistic functional interactions. AB - Using double in situ hybridization, we found extensive coexpression of dopamine D1 and D3 receptor (D1R and D3R) mRNAs in neurons of the island of Calleja major (ICjM) and ventromedial shell of nucleus accumbens (ShV), respectively. Thus, at least 79 and 63% of D3R mRNA-expressing neurons in ICjM and ShV also expressed the D1R mRNA. Coexpression of D1R and D3R mRNAs was found to occur in substance P (SP) mRNA-expressing neurons in both areas, suggesting SP mRNA as a marker of the activity of coexpressing neurons. Administration of SKF 38393, a D1R receptor agonist, increased c-fos mRNA in ICjM, whereas administration of quinpirole, a D2R/D3R agonist, decreased it; SCH 23390, a D1 R antagonist and nafadotride, a preferential D3R antagonist, given alone, had effects opposite to those of the corresponding agonists. These data indicate that basal c-fos expression in ICjM is maintained by endogenous dopamine acting tonically upon two receptor subtypes subserving opposite effects on the same cell. However, in ShV, whereas SKF 38393 also increased c-fos mRNA, quinpirole had no effect, a difference presumably reflecting the lower fraction of neurons coexpressing D1R and D3R in this area. In contrast, in ShV from reserpine-treated rats, SKF 38393 increased SP mRNA and quinpirole potentiated this effect. These contrasting interactions of D1R- and D3R-mediated signalling events, i.e. in either opposite or synergistic directions, most likely occurring at the single cell level, may serve to increase the dopamine response threshold of the target cells in ICjM and to maintain a strong tonic activity of ShV neurons. PMID- 9751141 TI - Glutamate receptor subunits GluR1 and GluR2/3 distribution shows reorganization in the human epileptogenic hippocampus. AB - The AMPA-type glutamate receptor subunits GluR1 and GluR2/3 were localized by immunohistochemistry with subunit-specific antibodies in hippocampi removed surgically from patients with temporal lobe epilepsy for the control of seizures. The flip and flop splice variants of the subunits were localized by in situ hybridization histochemistry with specific oligoprobes. In patient hippocampi that were not the seizure focus, the GluR1 subunit proteins were diffusely expressed on the dendrites of neurons in all regions. In contrast, in these same hippocampi, the GluR2/3 subunit proteins were expressed strongly on the soma and proximal dendrites of principal neurons in all regions. The flip variant of these subunits was localized in the hilus and fields of Ammon's Horn (CA), while the flop variants were prominent on the dentate granule cells. In the epileptogenic hippocampus, while immunoreactivity was decreased in all fields that showed neuronal loss, there was an increased expression of GluR1 on the dendritic excrescences on the proximal dendrites of hilar neurons and CA3 pyramidal neurons, as well as expression of GluR2/3 in hilar neuron excrescences. Electron microscopic examination confirmed that the GluR1 immunoreactivity was only in dendritic processes, particularly dense at the postsynaptic membranes. Such expression of GluR1 may provide for an enhanced glutamatergic response by these neurons. GluR2/3 was also significantly increased on the dendrites of dentate granule cells in the epileptogenic hippocampus and may provide some protection against excitotoxic injury by reducing calcium flux into neurons. PMID- 9751142 TI - Developmental changes in localization of NMDA receptor subunits in primary cultures of cortical neurons. AB - Immunoblot analysis, using antibodies against distinct N-methyl-D-aspartic acid (NMDA) receptor subunits, illustrated that the NR2A and NR2B subunit proteins have developmental profiles in cultured cortical neurons similar to those seen in vivo. NR1 and NR2B subunits display high levels of expression within the first week. In contrast, the NR2A subunit is barely detectable at 7 days in vitro (DIV) and then gradually increased to mature levels at DIV21. Immunocytochemical analysis indicated that NMDA receptor subunits cluster in the dendrites and soma of cortical neurons. Clusters of NR1 and NR2B subunits were observed as early as DIV3, while NR2A clusters were rarely observed before DIV10. At DIV18, NR2B clusters partially co-localize with those of NR2A subunits, but NR2B clusters always co-localize with those of NR1 subunits. Synapse formation, as indicated by the presence of presynaptic synaptophysin staining, was observed as early as 48 72 h after plating. However, in several neurons at ages less than DIV5 where synapses were scarce, NR2B and NR1 clusters were abundant. Furthermore, while NR2B subunit clusters were seen both at synaptic and extrasynaptic sites, NR2A clusters occurred almost exclusively in front of synaptophysin-labelled boutons. This result was supported by electrophysiological recording of NMDA-mediated synaptic activity [NMDA-excitatory postsynaptic currents (EPSCs)] in developing neurons. At DIV6, but not at DIV12, CP101, 606, a NR1/NR2B receptor antagonist, antagonized spontaneously occurring NMDA-EPSCs. Our data indicate that excitatory synapse formation occurs when NMDA receptors comprise NR1 and NR2B subunits, and that NR2A subunits cluster preferentially at synaptic sites. PMID- 9751143 TI - Evidence for a striatal NMDA receptor modulation of nigral glutamate release. A dual probe microdialysis study in the awake freely moving rat. AB - Dual probe microdialysis was employed to characterize dialysate glutamate levels from the substantia nigra pars reticulata of awake freely moving rats, and to test its sensitivity to alterations in striatal neurotransmission including striatal N-methyl-D-aspartic acid (NMDA) receptor stimulation and blockade. Intranigral perfusion with low (0.1 mM) Ca2+ medium (60 min) did not affect nigral glutamate levels, whereas intranigral perfusion with tetrodotoxin (10 microM, 60 min) increased nigral glutamate levels. Perfusion of KCI (100 mM, 10 min) in the dorsolateral striatum transiently stimulated nigral glutamate levels (maximal increase + 60%), whereas intrastriatal perfusion (60 min) with low Ca2+ medium and tetrodotoxin gradually increased nigral glutamate levels. Intrastriatal perfusion with NMDA (0.1-100 microM, 10 min) dose-dependently stimulated glutamate levels in the substantia nigra pars reticulata. The NMDA (1 microM)-induced increase in nigral glutamate release was transient and maximal (+60% within 20 min), whereas that for NMDA (10 microM) had a slow onset but was long lasting (+35% after 60 min). Lower (0.1 microM) and higher (100 microM) NMDA concentrations were ineffective. The effect of intrastriatal NMDA (1 microM) was prevented by coperfusion with MK-801 (1 microM). Intrastriatal MK-801 (10 microM) alone gradually increased glutamate levels up to +50% after 60 min of perfusion. The present results suggest that glutamate levels in the substantia nigra pars reticulata are sensitive to changes in neuronal transmission in the dorsolateral striatum, and that striatal NMDA receptors regulate nigral glutamate release in both a tonic and phasic fashion. PMID- 9751144 TI - Inhibition of ischaemic hippocampal neuronal death in primates with cathepsin B inhibitor CA-074: a novel strategy for neuroprotection based on 'calpain cathepsin hypothesis'. AB - Although Cornu Ammonis (CA) 1 neurons of the hippocampus are known to be vulnerable to transient ischaemia, the mechanism of ischaemic neuronal death is still unknown, and there are very few strategies to prevent neuronal death at present. In a previous report we demonstrated micro-calpain activation at the disrupted lysosomal membrane of postischaemic CA1 neurons in the monkey undergoing a complete 20 min whole brain ischaemia. Using the same experimental paradigm, we observed that the enzyme activity of the lysosomal protease cathepsin B increased throughout the hippocampus on days 3-5 after the transient ischaemia. Furthermore, by immunocytochemistry cathepsin B showed presence of extralysosomal immunoreactivity with specific localization to the cytoplasm of CA1 neurons and the neuropil of the vulnerable CA1 sector. When a specific inhibitor of cathepsin B, the epoxysuccinyl peptide CA-074 (C18H29N3O6) was intravenously administered immediately after the ischaemic insult, approximately 67% of CA1 neurons were saved from delayed neuronal death on day 5 in eight monkeys undergoing 20 min brain ischaemia: the extent of inhibition was excellent in three of eight and good in five of eight monkeys. The surviving neurons rescued by blockade of lysosomal activity, showed mild central chromatolysis and were associated with the decreased immunoreactivity for cathepsin B. These observations indicate that calpain-induced cathepsin B release is crucial for the development of the ischaemic neuronal death, and that a specific inhibitor of cathepsin B is of potential therapeutic utility in ischaemic injuries to the human CNS. PMID- 9751145 TI - Vasoactive intestinal peptide shortens both G1 and S phases of neural cell cycle in whole postimplantation cultured mouse embryos. AB - Vasoactive intestinal peptide, a trophic and mitogenic factor, stimulates growth in whole cultured mouse embryos. Inhibition of this growth function between embryonic days 9 and 11 induces growth retardation accompanied by severe microcephaly. In the present study, to determine the effects of this peptide on the different phases of the cell cycle of neural cells, embryonic day 9.5 cultured mouse embryos were cumulatively labelled with bromodeoxyuridine. Vasoactive intestinal peptide (10(-7)M) shortened S phase and G1 phase of neuroepithelial cells by 50% (4.8-2.4 h) and 58% (1.9-0.8 h), respectively, compared with controls. G2 and M phases were not modified by vasoactive intestinal peptide treatment. Total cell cycle length was consequently reduced by 43% (8.2-4.7 h) in vasoactive intestinal peptide treated embryos, compared with controls. In contrast, vasoactive intestinal peptide did not modify the rate of neuroepithelial cell death as assessed by the proportion of nuclei containing fragmented DNA. These data suggest that vasoactive intestinal peptide stimulates growth in premigratory stages of nervous system development by shortening S and G1 phases of the cell cycle and that S phase duration can be regulated by a physiological peptide. PMID- 9751146 TI - Delta9-tetrahydrocannabinol increases sequence-specific AP-1 DNA-binding activity and Fos-related antigens in the rat brain. AB - Delta-9-tetrahydrocannabinol (delta9-THC), the psychoactive principle of marijuana, has been shown to upregulate the mRNA levels of immediate-early genes in the rat brain. Using electrophoretic mobility-shift assay and one-dimensional Western blot, we here report that delta9-THC increases Activator protein-1 (AP-1) DNA-binding and Fos-related antigen activity in discrete areas of the rat brain. One hour after the intraperitoneal administration of delta9-THC at a dose of 10 or 15 mg/kg, AP-1 DNA-binding activity in the nucleus accumbens increased by 33 and 49%, respectively, while Western blot showed an increase in both c-Fos, FosB, Fra-1 (Fos-related antigen) and Fra-2. In the cingulate cortex and caudate putamen, delta9-THC significantly increased AP-1 DNA-binding activity only at the highest dose used (57 and 71%, respectively). While in the caudate-putamen the increase in AP-1 DNA binding was mainly due to an elevation of the c-Fos and FosB proteins, the same phenomenon depended on the FosB, Fra-1 and Fra-2 peptides in the cingulate cortex. The effect of delta9-THC on the AP-1 DNA binding and the Fos-related antigens in the nucleus accumbens was blocked by the specific cannabinoid antagonist SR141716 A (3 mg/kg i.p.). delta9-THC failed to modify Specificity protein 1 (Sp1) DNA-binding activity. The results indicate that delta9-THC activates gene coding for AP-1 DNA-binding proteins by acting on cannabinoid receptors, and induces a different transcriptional program on the early-immediate gene of the Fos family, in different areas in the rat brain, suggesting that this mechanism might be involved in the central actions of cannabinoids. PMID- 9751147 TI - Induction of neurosteroid synthesis by NMDA receptors in isolated rat retina: a potential early event in excitotoxicity. AB - Here we investigated the possible regulation of neurosteroidogenesis by N-methyl D-aspartic acid (NMDA) receptor activation and addressed the hypothesis that neurosteroid synthesis may be involved in acute excitotoxicity. In the isolated retina, exposure to NMDA modified pregnenolone and pregnenolone sulphate formation. This effect was dose and time dependent, the synthesis being increased by relatively moderate NMDA doses (1-100 microM) within 30 min exposure and reduced to its control value by 60 min or by raising drug concentrations. NMDA stimulated neurosteroid synthesis was blocked by (+)-5-methyl-10,11-dihydro-5H dibenzo[a,d]cyclo-hepten-5,10-imine hydrogen maleate (MK-801) and 3(2 carboxypiperazine-4-yl)propyl-1-phosphonic acid (CPP), depended on extracellular calcium and reproduced by glutamate. Lactate dehydrogenase (LDH) release and morphological analysis revealed that retinal cell viability was not significantly affected after 30 min exposure to 50 microM NMDA, but severe cell damage occurred by 60 min. When the GABAA (gamma-aminobutyric acid) receptor agonist muscimol (1 1000 microM), known to activate retinal neurosteroidogenesis, was added together with NMDA, no additional increase in neurosteroid synthesis was observed, and NMDA-induced LDH release remained unchanged. However, exposure to a high concentration of muscimol alone (500 microM) provoked a similar degree of toxicity to NMDA. By contrast, bicuculline abolished the increase in neurosteroidogenesis and LDH release. Similarly, pretreatment with R (+)-p aminoglutethimide (AMG), an inhibitor of cholesterol side-chain cleavage cytochrome P450, attenuated acute retinal cell damage. The inhibitory nature of AMG on NMDA-stimulated neurosteroidogenesis was confirmed in the observation that drug treatment reduced pregnenolone content and did not affect the bindings of [3H] MK-801 and [3H] muscimol. The results demonstrate that NMDA receptors regulate neurosteroidogenesis through a transneuronal mechanism, which implies GABAA receptor activation. The early NMDA-mediated stimulation of neurosteroid synthesis seems to play a critical role in acute excitotoxicity; consequently, its inhibition is likely to delay neuronal cell death. PMID- 9751148 TI - Direct evidence of cubic difference tone propagation by intracochlear acoustic pressure measurements in the guinea-pig. AB - The fine tuning mechanisms involved in the normal processing of sound in the cochlea are non-linear, hence combination tones are generated inside the cochlea when a pair of low-level pure tones with neighbouring frequencies f1 and f2 is used as a stimulus. Their detection as sounds in the ear canal proves that they undergo backward propagation in the cochlea and through the middle ear, and the non-invasive measurement of the combination tone at 2f1-f2, called the cubic difference tone (CDT), has become a routine method of monitoring cochlear function. In order to gain information on the hypothetical places where CDTs are generated, on their intracochlear levels and propagation velocities, direct measurements of CDT pressure waves were carried out in scala vestibuli and tympani of the first and second turn of the guinea-pig cochlea. Cubic difference tones at 2f1-f2 varied from 0.75 to 9 kHz and were measured with a miniature piezoresistive transducer. Its high sensitivity allowed the detection of CDTs whenever their levels exceeded 5 dB SPL in the ear canal, i.e. 40 dB SPL (re: 20 microPa) inside the cochlea. The levels of CDTs were similar in scala vestibuli of the first and second turn. Phase comparisons between measurements at 2f1-f2 in the first and second turn allowed determination of the place where the CDT phase was minimum. It provided an estimation of the generation site of the CDT, which appeared to be close to the place tuned to f2 for stimulus levels lower than 70 dB SPL. Forward and backward travel times from one turn to the other were assessed at several frequencies, and both values were shorter than 0.2 ms. In contrast, the overall 'round-trip' delay of CDTs, measured in the ear canal, was about five times larger, suggesting that local filtering processes rather than propagation delays account for the overall CDT delay. PMID- 9751149 TI - Initially expressed early rat embryonic GABA(A) receptor Cl- ion channels exhibit heterogeneous channel properties. AB - We have studied the earliest expression of GABA-induced CI- channels in the rat embryonic dorsal spinal cord (DSC) using in situ hybridization, immunocytochemistry, flow cytometry and electrophysiology. At embryonic day 13 (E13) cells in the dorsal region are still proliferating. In situ hybridization consistently showed transcripts encoding only three GABAA receptor subunits (alpha4, beta1 and gammal); immunocytochemistry both in tissue sections and in acutely isolated cells in suspension demonstrated the expression of the corresponding proteins and also revealed staining for other subunits (alpha2, alpha3, beta3, gamma2). In patch-recordings performed in cells acutely isolated from the dorsal cord, responses to GABA were detected in 356 out of 889 cells. GABA-evoked responses, which often displayed the opening of a few channels, were mediated by CI- ions, were inhibited by bicuculline and picrotoxin, and potentiated by benzodiazepines. Taken together, these observations indicate that CI- channels likely involve GABAA type receptors. Fluctuation analysis revealed channel kinetics consisting of three exponential components (Ts: approximately 1,9 and 90 ms) and a wide variety of inferred unitary conductance values, ranging between 4 and 40 pS. A comparison of these results with observations in other, later embryonic cell types and recombinant receptors suggests that most of the earliest E13 DSC GABAA receptors may include alpha3 subunit. These GABAA receptor Cl- channels may be activated physiologically as both GABA synthesizing enzymes and GABA are present in the E13 dorsal cord. PMID- 9751150 TI - Enkephalin-containing interneurons are specialized to innervate other interneurons in the hippocampal CA1 region of the rat and guinea-pig. AB - Enkephalins are known to have a profound effect on hippocampal inhibition, but the possible endogenous source of these neuropeptides, and their relationship to inhibitory interneurons is still to be identified. In the present study we analysed the morphological characteristics of met-enkephalin-immunoreactive cells in the CA1 region of the rat and guinea-pig hippocampus, their coexistence with other neuronal markers and their target selectivity at the light and electron microscopic levels. Several interneurons in all subfields of the hippocampus were found to be immunoreactive for met-enkephalin. In the guinea-pig, fibres arising from immunoreactive interneurons were seen to form a plexus in the stratum oriens/alveus border zone, and basket-like arrays of boutons on both enkephalin immunoreactive and immunonegative cell bodies in all strata. Immunoreactive boutons always established symmetric synaptic contacts on somata and dendritic shafts. Enkephalin-immunoreactive cells co-localized GABA, vasoactive intestinal polypeptide and calretinin. Postembedding immunogold staining for GABA showed that all the analysed enkephalin-immunoreactive boutons contacted GABAergic postsynaptic structures. In double-immunostained sections, enkephalin-positive axons were seen to innervate calbindin D28k-, somatostatin-, calretinin- and vasoactive intestinal polypeptideimmunoreactive cells with multiple contacts. Based on these characteristics, enkephalin-containing cells in the hippocampus are classified as interneurons specialized to innervate other interneurons, and represent a subset of vasoactive intestinal polypeptide- and calretinin containing cells. The striking match of ligand and receptor distribution in the case of enkephalin-mediated interneuronal communication suggests that this neuropeptide may play an important role in the synchronization and timing of inhibition involved in rhythmic network activities of the hippocampus. PMID- 9751151 TI - Functional Ca2+ and Na+ channels on mouse Schwann cells cultured in serum-free medium: regulation by a diffusible factor from neurons and by cAMP. AB - Regulation of expression of functional voltage-gated ion channels for inward currents was studied in Schwann cells in organotypic cultures of dorsal root ganglia from E19 mouse embryos maintained in serum-free medium. Of the Schwann cells that did not contact axons, 46.5% expressed T-type Ca2+ conductances (ICaT). Two days or more after excision of the ganglia, and consequent disappearance of neurites, ICaT were detectable in only 10.9% of the cells, and the marker 04 disappeared. On Schwann cells deprived of neurons, T- (but not L-) type Ca2+ conductances were re-induced by weakly hydrolysable analogues of cAMP, and by forskolin (an activator of adenylyl cyclase) after long-term treatment (4 days). With CPT cAMP (0.1-2 mM), 8Br cAMP, db cAMP or forskolin (0.01 or 0.1 mM), the proportion of cells with ICaT was not significantly different from the proportion in the cultures with neurons. These agents also induced expression in some cells of tetrodotoxin-resistant Na+ currents, which were rarely induced by neurons, but 04 was not re-induced by cAMP analogue treatments that re-induced ICaT. Inward currents (Ba2+ or Na+) were partly restored (P < 0.05) on Schwann cells cultured for 6-7 days beneath a filter bearing cultured neurons. In contrast, addition of neuron-conditioned medium was ineffective. The results suggest that neurons activate, via diffusible and degradable factors, a subset of Schwann cell cAMP pathways leading to expression of IcaT, and activate additional non-cAMP pathways that lead to expression of 04. PMID- 9751153 TI - Spectral characteristics of sleep EEG in chronic insomnia. AB - To determine whether the spectral characteristics of the sleep electroencephalogram (EEG) of insomniacs differ from that of healthy subjects, we compared in each of the first four non-rapid eye movement (NREM) and rapid eye movement (REM) episodes: (a) the time courses of absolute power, averaged over the subjects in each group, for the delta, theta, alpha, sigma and beta frequency bands; (b) the relationship between these time courses; and (c) the overnight trend of integrated power in each frequency band. The results show that NREM power, for all frequencies below the beta range, has slower rise rates and reaches lower levels in the insomniac group, whereas beta power is significantly increased. In REM, insomniacs show lower levels in the delta and theta bands, whereas power in the faster frequency bands is significantly increased. Thus, the pathophysiology of insomnia is characterized not only by the generally acknowledged slow wave deficiency, but also by an excessive hyperarousal of the central nervous system throughout the night, affecting both REM and NREM sleep. This hyperarousal is interpreted in terms of the neuronal group theory of sleep which provides a possible explanation for the discrepancies observed between subjective impressions and objective measures of sleep. Also, it is suggested that the progressive hyperpolarization of the thalamocortical neurons as sleep deepens is slower in the patient population and that this may explain the observed slow wave deficiency. The homeostatic control of slow wave activity, on the other hand, would appear to be intact in the patient population. PMID- 9751152 TI - Antagonists-resistant calcium currents in rat embryo motoneurons. AB - Ca2+ channels diversity of cultured rat embryo motoneurons was investigated with whole-cell current recordings. In 5-20 mM Ba2+, the whole-cell currents were separated in low- (LVA) and high-voltage-activated (HVA) current. The LVA current was evident since the first day in culture, while the HVA component was small and increased with time. Recordings after 4 days revealed approximately 20% L-, approximately 45% N- and approximately 35% P- and R-type currents. P-type currents were revealed only in 40% of motoneurons, in which 20-200 nM omega-Aga IVA caused 20% irreversible block of total current. The remaining 60% of cells were insensitive even to higher doses of the toxin (500 nM in 5 mM Ba2+), suggesting weak expression and heterogeneous distribution of P-type channels compensated by high densities of HVA Ca2+ channels resistant to all the antagonists (R-type). A significant residual current could also be resolved after prolonged applications of 5 microM omega-CTx-MVIIC, which allowed separation of N and P-type currents by the distinct onset of toxin block. The antagonists resistant current reveals biophysical characteristics typical of HVA channels, but distinct from the alphaE channel. The current activates around -20 mV in 20 mM Ba2+; inactivates slowly and independently of Ca2+; is blocked by low [Cd2+] and high [Ni2+]; and is larger with Ba2+ than Ca2+. The uncovered R-type calcium current can account for part of the presynaptic Ca2+ current controlling neurotransmitter release at the mammalian neuromuscular junction whose activity is resistant to DHP-and omega-CTx-GVIA, and displays anomalous sensitivity to omega-Aga-IVA and omega-CTx-MVIIC. PMID- 9751154 TI - The expression of Huntingtin-associated protein (HAP1) mRNA in developing, adult and ageing rat CNS: implications for Huntington's disease neuropathology. AB - Using radioactive in situ hybridization, we have mapped the expression of Huntingtin-associated protein (HAP1) mRNA in rat brain at developmental stages (E12-E19, PO-P21), in adult rats (3 months) and in 'aged' (19-21 months) rats. Using two pairs of 45mer oligonucleotide probes specific for HAP1A and a probe which recognizes regions of both the HAP1A and HAP1B mRNA sequences (panHAP1), we find that the expression of HAP1 mRNA is specific to the CNS and restricted predominantly to anatomically connected limbic structures, particularly the amygdala (medial and corticomedial nuclei), the hypothalamus (arcuate, preoptic, paraventricular and lateral hypothalamic area), bed nucleus of the stria terminalis (BNST) and the lateral septal nuclei. HAP1 mRNA was detected in embryos at E12 and displayed a prevalent distribution in the developing limbic structures by E15. In aged, 19-21-months-old, rats there is a downregulation of HAP1 mRNA expression across all CNS loci where HAP1 was previously abundant. The lowest levels of HAP1 mRNA expression corresponded with the areas of greatest pathological cell loss in Huntington's disease (HD); the caudate putamen, globus pallidus and neocortex. These observations support the suggestion that HAP1 plays an important role in the neuropathology of HD. PMID- 9751155 TI - Local nitric oxide synthase activity in a model of neuropathic pain. AB - A local inflammatory reaction may play an important role in the development of neuropathic pain following peripheral nerve injury. One important participant in the inflammatory response of injured peripheral nerve may be nitric oxide (NO). In this work, we examined physiological and morphological evidence for nitric oxide synthase (NOS) activation in the chronic constriction injury model of neuropathic pain in rats. Physiological evidence of local NO action was provided by studying NO-mediated changes in local blood flow associated with the injury site. Immunohistochemistry was used to localize isoforms of NOS that might generate NO. Sciatic nerve injury associated with behavioural evidence of neuropathic pain had substantial rises in local blood flow. The NOS inhibitor NG nitro-L-arginine methyl ester (L-NAME), but not NG-nitro-D-arginine methyl ester (D-NAME), reversed the hyperaemia in a dose-dependent fashion proximal to the constriction at 48 h and distally at 14 days post-operation when applied systemically or topically. Aminoguanidine, a NOS inhibitor with relatively greater selectivity for the inducible NOS (iNOS) isoform, reversed nerve hyperaemia distal to the constriction only at 14 days. NOS-like immunoreactivity of the neuronal and endothelial isoforms was identified just proximal to the constriction at 48 h. iNOS-like immunoreactivity was observed at 7 and 14 days at the constriction and distal sites, respectively. This work provides evidence for local NOS expression and NO action in the chronic constriction injury model of neuropathic pain. NO has local physiological actions that include vasodilatation of microvessels and that may be important in the development of pain sensitivity. PMID- 9751156 TI - The response of cat visual cortex to flicker stimuli of variable frequency. AB - We examined the possibility that neurons or groups of neurons along the retino cortical transmission chain have properties of tuned oscillators: To this end, we studied the resonance properties of the retino-thalamo-cortical system of anaesthetized cats by entraining responses with flicker stimuli of variable frequency (2-50 Hz). Responses were assessed from multi-unit activity (MUA) and local field potentials (LFPs) with up to four spatially segregated electrodes placed in areas 17 and 18. MUA and LFP responses were closely related, units discharging with high preference during LFP negativity. About 300 ms after flicker onset, responses stabilized and exhibited a highly regular oscillatory patterning that was surprisingly similar at different recording sites due to precise stimulus locking. Fourier transforms of these steady state oscillations showed maximal power at the inducing frequency and consistently revealed additional peaks at harmonic frequencies. The frequency-dependent amplitude changes of the fundamental and harmonic response components suggest that the retino-cortical system is entrainable into steady state oscillations over a broad frequency range and exhibits preferences for distinct frequencies in the theta- or slow alpha-range, and in the beta- and gamma-band. Concomitant activation of the mesencephalic reticular formation increased the ability of cortical cells to follow high frequency stimulation, and enhanced dramatically the amplitude of first- and second-order harmonics in the gamma-frequency range between 30 and 50 Hz. Cross-correlations computed between responses recorded simultaneously from different sites revealed pronounced synchronicity due to precise stimulus locking. These results suggest that the retino-cortical system contains broadly tuned, strongly damped oscillators which altogether exhibit at least three ranges of preferred frequencies, the relative expression of the preferences depending on the central state. These properties agree with the characteristics of oscillatory responses evoked by non-temporally modulated stimuli, and they indicate that neuronal responses along the retino-cortical transmission chain can become synchronized with precision in the millisecond range not only by intrinsic interactions, but also by temporally structured stimuli. PMID- 9751157 TI - Motor stimulant effects of caffeine in 6-hydroxydopamine-lesioned rats are dependent on previous stimulation of dopamine receptors: a different role of D1 and D2 receptors. AB - Caffeine has been reported to induce contralateral rotational behaviour in rats bearing a unilateral 6-hydroxydopamine lesion of the dopaminergic nigrostriatal pathway. In order to define the role of dopamine receptors in the mediation of this behaviour, we have evaluated the influence of previous exposure to a dopamine receptor agonist and the importance of the time elapsed from the 6 hydroxydopamine lesion on the rotational behaviour induced by caffeine. Separate groups of rats lesioned with 6-hydroxydopamine 2 weeks previously were exposed to four administrations of the D1/D2 receptor agonist apomorphine (0.3 mg/kg s.c.) (primed) or vehicle (drug-naive). Three days later, all rats received caffeine (30 mg/kg s.c.). Drug-naive 6-hydroxydopamine-lesioned rats did not rotate in response to caffeine, while rats primed with apomorphine rotate contralaterally in response to caffeine. When apomorphine priming was paired to the same environment (hemispherical bowls) where rats received caffeine, rotational behaviour was significantly higher than that obtained in rats primed in an unpaired environment (cylinders). Repeated priming with the D2/D3 receptor agonist quinpirole (0.2 mg/kg s.c.) induced a totally context-dependent contralateral rotation in response to caffeine, while caffeine contralateral rotation was not dependent from the context after repeated priming with the D1 agonist SKF 38393 [1-phenyl-2,3,4,5-tetrahydro-(1 H)-3-benzazepine-7,8-diol hydrochloride, 3 mg/kg s.c.]. Caffeine-mediated contralateral rotation was also evaluated in rats lesioned with 6-hydroxydopamine 12 weeks previously and exposed to four administrations of apomorphine or vehicle. As for rats repeatedly exposed to vehicle or apomorphine 2 weeks after 6-hydroxydopamine lesioning, caffeine failed to induce contralateral rotation in drug-naive rats, while it did induce a partially context-dependent contralateral rotation in apomorphine-primed rats. Different from rats receiving apomorphine priming 2 weeks after 6-hydroxydopamine lesioning, in 12 week-lesioned rats, caffeine also induced contralateral rotation after one priming with apomorphine (0.3 mg/kg s.c.), a condition which fails to induce context-dependent rotation. Administration of selective antagonists of A1 (8-cyclopentyl-1,3-dipropylxanthine), (DPCPX) or A2A (5-amino-2-(2-furyl)-7-(3 phenylpropyl)-pyrazolo[4,3-e]-1 ,2,4-triazolo[5c]pirimidine), (SCH 58261) adenosine receptors failed to induce contralateral rotation either alone or in combination in 12 week-6-hydroxydopamine-lesioned rats repeatedly primed with apomorphine. All together, the results indicate that: (i) caffeine does not induce any contralateral rotation in drug-naive 6-hydroxydopamine-lesioned rats; (ii) priming with a dopamine agonist enables caffeine to induce contralateral rotation, this rotation is, however, context independent only after priming with a selective D1 agonist; (iii) contralateral rotation in response to caffeine is dependent on the time from the 6-hydroxydopamine lesion; (iv) blockade of A1 and A2A adenosine receptors with selective antagonists does not induce contralateral rotational behaviour in 6-hydroxydopamine-lesioned rats. PMID- 9751158 TI - Melanocortin receptors and delta-opioid receptor mediate opposite signalling actions of POMC-derived peptides in CATH.a cells. AB - The locus coeruleus is innervated by proopiomelanocortin (POMC)-derived peptide immunoreactive fibres. The biological effects of ( melanocyte-stimulating hormone (aMSH) and [-endorphin on second messengers (cAMP, inositol phosphates) and gene transcription were studied in the locus cceruleus-derived cell line CATH.a. RT PCR analysis revealed the presence of four MSH receptor subtypes (1, 3, 4 and 5). Activation of these receptors by diacetyl alphaMSH stimulated cAMP accumulation in a dose-dependent manner (EC50: 4 x 10(-9) M). Diacetyl alphaMSH stimulated transcription from reporter genes driven by the c-fos or tyrosine hydroxylase promoter. This effect was abolished when protein kinase A was inactivated with a dominant inhibitory mutant. RT-PCR analyses revealed the presence of delta-, but not mu-and kappa-opioid receptor. Pharmacological analysis showed that beta endorphin (EC50: 2.5 x 10(-8)M), but not N-acetyl beta-endorphin, antagonized the biological effect of diacetyl alphaMSH on cAMP production and gene transcription. Since N-acetylation regulates the biological activity of alphaMSH and beta endorphin in an opposite manner, we propose a model where the rate of secretion dictated by the bioelectric activity of the presynaptic neuron modulates POMC derived peptide maturation and the resulting biological signal sensed by the postsynaptic plate. PMID- 9751159 TI - Mesolimbic dopaminergic mechanisms underlying individual differences in sugar consumption and amphetamine hyperlocomotion in Wistar rats. AB - Individual differences within strains of rats have been demonstrated for dopamine mediated behaviours and responses to dopaminergic drugs. Differences in mesolimbic dopamine function may underlie individual differences in some of these behaviours, including sugar consumption and amphetamine hyperlocomotion. The present study addressed two potential mechanisms for these differences in dopamine-mediated behaviours. The possibility of functional differences in dopamine receptor subtypes was tested in LOW and HIGH sugar feeders. LOW and HIGH feeders did not differ in their response to the partial D1 agonist SKF-38393. The highest dose (2.5 mg/kg) of the D2 agonist quinpirole stimulated locomotor activity to a greater degree in a subset of HIGH sugar feeders as compared with LOW feeders. All doses of amphetamine induced a greater locomotor response in HIGH feeders as compared with LOW feeders, and HIGH feeders exhibited higher levels of extracellular dopamine in the nucleus accumbens than LOW feeders following exposure to sugar and treatment with amphetamine. These results support the interpretation that LOW and HIGH feeders exhibit differences in presynaptic nucleus accumbens dopamine function that account for the expression of individual differences in sugar consumption and response to amphetamine treatments. A subset of HIGH feeders may also exhibit greater D2 receptor function. PMID- 9751160 TI - NMDA-induced superoxide production and neurotoxicity in cultured rat hippocampal neurons: role of mitochondria. AB - Excitotoxic mechanisms are believed to be involved in the death of neurons after trauma, epileptic seizures and cerebral ischaemia. We investigated the role of mitochondrial superoxide production in excitotoxic cell death of cultured rat hippocampal neurons. Brief exposure to the selective glutamate agonist N-methyl-D aspartate (NMDA; 100-300 microM, 10 min) induced significant neuronal death, which was sensitive to cycloheximide (1 microM) and the caspase-1 inhibitor, acetyl-Tyr-Val-Ala-Asp-chloromethylketone (10 microM). Intracellular superoxide production was monitored semiquantitatively on sister cultures from the same platings using the oxidation-sensitive probe, hydroethidine. Brief exposures to toxic NMDA concentrations induced significant increases in superoxide production which correlated with the degree of neuronal injury. However, subtoxic NMDA exposures also produced moderate, yet statistically significant increases in superoxide production. Both NMDA-induced superoxide production and neurotoxicity were reduced by inhibition of mitochondrial electron transport using either sodium cyanide (1 mM), or a combination of rotenone (2 microM) and oligomycin (2 microM). The mitochondrial uncoupler carbonyl cyanide p-trifluoromethoxy phenylhydrazone (FCCP, 1 microM) mimicked the effect of NMDA on mitochondrial superoxide production. Both NMDA-induced superoxide production and neurotoxicity were potentiated by FCCP (1 microM). Exposure to FCCP alone (1-10 microM, 10 min), however, failed to produce any toxicity. Our data suggest that mitochondrial superoxide production per se is not sufficient to trigger the degeneration of cultured hippocampal neurons, but that manipulation of mitochondrial activity alters NMDA-induced superoxide production and neurotoxicity. PMID- 9751162 TI - Hyperpolarization induces a rise in intracellular sodium concentration in dopamine cells of the substantia nigra pars compacta. AB - We investigated the effect of changes in membrane-voltage on intracellular sodium concentration ([Na+]i) of dopamine-sensitive neurons of the substantia nigra pars compacta in a slice preparation of rat mesencephalon. Whole-cell patch-clamp techniques were combined with microfluorometric measurements of [Na+]i using the Na+-sensitive probe, sodium-binding benzofuran isophthalate (SBFI). Hyperpolarization of spontaneously active dopamine neurons (recorded in current clamp mode) caused the cessation of action potential firing accompanied by an elevation in [Na+]i. In dopamine neurons voltage-clamped at a holding potential of -60 mV elevations of [Na+]i were induced by long-lasting (45-60 s) voltage jumps to more negative membrane potentials (-90 to -120 mV) but not by corresponding voltage jumps to -30 mV. These hyperpolarization-induced elevations of [Na+]i were depressed during inhibition of I(h), a hyperpolarization-activated inward current, by Cs+. Hyperpolarization-induced elevations in [Na+]i might occur also in other cell types which express a powerful I(h) and might signal lack of postsynaptic activity. PMID- 9751161 TI - Identification and characterization of a novel member of the fibroblast growth factor family. AB - A new member of the fibroblast growth factor (FGF) family, FGF-13, has been molecularly cloned as a result of high throughput sequencing of a human ovarian cancer cell library. The open reading frame of the novel human gene (1419 bp) encodes for a protein of 216 a.a. with a molecular weight of 22 kDa. The FGF-13 sequence contains an amino-terminal hydrophobic region of 23 a.a. characteristic of a signal secretion sequence. FGF-13 is most homologous, 70% similarity at the amino acid level, to FGF-8. Northern hybridization analysis demonstrated prominent expression of FGF-13 in human foetal and adult brain, particularly in the cerebellum and cortex. In proliferation studies with BaF3 cells, FGF-13 preferentially activates cell clones expressing either FGF receptor variant, 3 IIIc or 4. The signal transduction pathways of FGF-13 and FGF-2 were compared in rat hippocampal astrocytes. The two FGFs induce an equivalent level of tyrosine phosphorylation of mitogen-activated protein kinase (MAPK) and c-raf activation. However, FGF-13 is more effective than FGF-2 in inducing the phosphorylation of phospholipase C-gamma (PLC-gamma). Treatment of neuronal cultures from rat embryonic cortex with FGF-13 increases the number of glutamic acid decarboxylase immunopositive neurons, the level of high-affinity gamma-aminobutyric acid (GABA) uptake, and choline acetyltransferase enzyme activity. The GABAergic neuronal response to FGF-13 treatment is rapid with a significant increase occurring within 72 h. We have identified a novel member of the FGF family that is expressed in the central nervous system (CNS) and increases the number as well as the level of phenotypic differentiation of cortical neurons in vitro. PMID- 9751163 TI - Serotonin is not synthesized, but specifically transported in the neurons of the hypothalamic dorsomedial nucleus. AB - A small group of neurons in the hypothalamic dorsomedial nucleus (DMN) have been reported to contain serotonin after pharmacological treatments enhancing brain serotonin levels. This study aimed at elucidating whether these neurons are able to synthesize serotonin de novo, and whether they possess a specific serotonin transport mechanism. Serotonin content in these neurons was raised by administration of L-tryptophan and pargyline. Double immunostaining for serotonin and tryptophan hydroxylase (TpOH), the serotonin synthesizing enzyme, revealed that none of the serotonin-containing neuronal somata expressed TpOH. Intracerebroventricular colchicine treatment did not result in TpOH-IR in these neurons. Fluoxetine, a specific serotonin transport inhibitor, prevented the accumulation of serotonin in these neurons. The present results thus indicate that the serotonin-containing DMN neurons are not able to synthesize serotonin. Instead, they take up exogenous serotonin via a specific serotonin transport mechanism. As serotonin and DMN are associated with various physiological functions, such as regulation of food intake and modulation of fear and anxiety, the mechanisms revealed in the present study may participate in these clinically important brain functions. PMID- 9751164 TI - The making of neurexins. AB - Neurexins are neuronal cell-surface proteins with up to thousands of isoforms. These isoforms are generated by alternative splicing of transcripts from six promoters in three genes. The structure of neurexins resembles cell-surface receptors with a modular architecture suggestive of a sequential assembly during evolution. Neurexins probably perform multiple functions in the brain. They participate in intercellular junctions in which beta-neurexins tightly bind to a second class of neuronal cell-surface receptors called neuroligins. Intracellularly, the neurexin/neuroligin junction is bound by CASK on the neurexin side and PSD95 on the neuroligin side. CASK and PSD95 are homologous membrane-associated guanylate kinases that bind to the neurexin/neuroligin junction via PDZ domains, creating an asymmetric junction (neurexin/neuroligin) with similar intracellular binding partners. In addition to a function as cell adhesion molecules, neurexins may also serve as a signalling receptor, because a class of ligands for alpha-neurexins called neurexophilins is similar to peptide hormones. Finally, at least one neurexin isoform, neurexin Ialpha, represents a high-affinity receptor for alpha-latrotoxin, which is a potent excitatory neurotoxin. Thus, neurexins constitute a large family of neuronal receptors that may be involved in multiple interactive functions between neurons. PMID- 9751165 TI - Molecular cloning and characterization of the rat P2Y4 receptor. AB - Degenerate PCR was used to amplify DNAs encoding members of the P2Y receptor family from rat brain RNA. A full-length sequence obtained for one novel clone (R5) contained an intronless open reading frame that encoded a polypeptide of 361 amino acids, sharing 84% sequence identity with the human P2Y4 receptor. When R5 was stably expressed in Jurkat cells, calcium fluxes resulting from stimulation of the receptor showed that UDP, ADP, 2-methylthio-ATP, and diadenosine tetraphosphate were inactive, whereas UTP and ATP were both full agonists with similar potency. At the human receptor, ATP has significantly lower potency than UTP. The R5 transcript was not detected in brain by northern hybridization. Therefore, its tissue distribution was assessed by PCR, and the mRNA was found to be widely distributed at a low abundance, being present in brain, spinal cord, and a variety of peripheral organs. Localization of the receptor transcript in adult rat brain sections by in situ hybridization indicated that it is expressed at highest levels in the pineal gland and ventricular system. It is presumed that R5 is a species orthologue of the human P2Y4 receptor but with this significant difference in agonist pharmacology. PMID- 9751166 TI - Identification and characterization of potential cis-regulatory elements governing transcriptional activation of the rat tyrosine hydroxylase gene. AB - Tyrosine hydroxylase (TH) catalyzes the conversion of L-tyrosine to 3,4-dihydroxy L-phenylalanine, which is the first and rate-limiting step in catecholamine biosynthesis. We have previously shown that the cyclic AMP response element (CRE), an essential promoter element for both basal and cyclic AMP-inducible TH transcription, activates the promoter activity in a distance-dependent manner. To identify further cis-regulatory elements controlling TH gene expression, we analyzed the potential regulatory sequences by several approaches. First, using transient transfection assays, we examined the cell-specific promoter activities of TH-reporter gene constructs and a dopamine beta-hydroxylase (DBH)-reporter construct containing the 5' upstream sequences of the rat TH and human DBH genes, respectively, that had been shown to direct tissue-specific reporter expression in transgenic mice experiments. Second, DNase I footprinting analysis of the 503 bp proximal area of the rat TH gene identified seven footprinted regions that encompass the putative cis-regulatory motifs, including the CRE (domain 1), Sp1 (domain III), Octamer (domain IV), AP1 (domain V), AP2 (domain VI), and two potentially novel sequence motifs (domains II and VII). Footprinting patterns at these sites by nuclear proteins from TH-positive and -negative cell lines appeared to be similar. Third, site-directed mutagenesis demonstrates that domain III, but not domain II, critically contributes to the TH promoter activity. Furthermore, electrophoretic mobility shift, competition, and supershift assays demonstrate that domain III is an authentic Sp1 site and that the transcription factor Sp1 interacts with it. This and previous results suggest that the CRE and Sp1 site may synergistically activate TH transcription in a promoter context dependent manner. PMID- 9751167 TI - Cloning and characterization of a novel form of tyrosine hydroxylase from the human parasite, Schistosoma mansoni. AB - Catecholamines such as dopamine and noradrenaline play important roles as neuromuscular transmitters and modulators in all parasitic helminths, including the human parasite, Schistosoma mansoni. We have cloned a novel S. mansoni tyrosine hydroxylase (SmTH) cDNA that shows high homology to mammalian tyrosine hydroxylase, the enzyme that catalyzes the first and rate-limiting step in the biosynthesis of catecholamines. Two subsets of SmTH transcripts were identified, one of which carries the S. mansoni spliced-leader (SL) sequence at its 5' end, whereas the other does not appear to be trans-spliced to the S. mansoni SL. The two types of SmTH transcripts encode the same protein of 465 amino acids and a predicted size of 54 kDa. Expression of SmTH as an N-terminal histidine fusion protein in Escherichia coli produced an active enzyme that was purified approximately 52-fold to apparent homogeneity and had a final specific activity of 0.78 micromol/min/mg. The purified enzyme was found to have the same absolute requirement for a tetrahydrobiopterin cofactor and the same sensitivity to inhibition by high concentrations of the substrate, tyrosine, as the mammalian enzyme. Purified SmTH also showed characteristic inhibition by catecholamine products, although the sensitivity to product inhibition was lower than that of the mammalian enzyme. This evidence indicates that SmTH encodes a functional tyrosine hydroxylase that has catalytic properties similar to those of the mammalian host's enzyme but may differ in its properties of regulation. This first demonstration of tyrosine hydroxylase in a parasitic helminth further suggests that the parasites have the enzymatic capacity to synthesize catecholamines endogenously. PMID- 9751168 TI - Migration behavior of rodent granule neurons in the presence of antibody to the 4C5 antigen. AB - We have reported the production of monoclonal antibody 4C5, which recognizes a cell surface antigen, the 4C5 antigen, involved in granule cell migration processes. In the present study, we investigated in a more precise manner the role of the 4C5 antigen in the different types of granule cell migrations that take place during cerebellar development. When cerebellar explant cultures derived from 10-day-old rats were performed for 2 days in the presence of monoclonal antibody 4C5, vertical granule cell migration, occurring in the presence of glia, was not significantly inhibited. In contrast, when monoclonal antibody 4C5 was included in the medium of microexplant cultures derived from 4 day-old mice and maintained for 4 days in vitro, granule cell migrations that occurred both parallel and perpendicular to the neurite bundles that were free of glia were inhibited. Moreover, a stronger inhibitory effect of the antibody was observed on migration perpendicular to the neurite bundles compared with the parallel type of migration. Our results indicate that the 4C5 antigen differentially affects the different developmental stages and types of granule cell migration during rodent cerebellar development. PMID- 9751169 TI - Ca2+-mediated activation of c-Jun N-terminal kinase and nuclear factor kappa B by NMDA in cortical cell cultures. AB - We examined the possibility that c-Jun N-terminal kinase (JNK) and nuclear factor kappaB (NF-kappaB) might be involved in intracellular signaling cascades that mediate NMDA-initiated neuronal events. Exposure of cortical neurons to 100 microM NMDA induced activation of JNK within 1 min. Activity of JNK was further increased over the next 5 min and then declined by 30 min. Similarly, ionomycin, a selective Ca2+ ionophore, induced activation of JNK. The NMDA-induced activation of JNK was abrogated in the absence of extracellular Ca2+, suggesting that Ca2+ entry is necessary and sufficient for the JNK activation. Immunohistochemistry with anti-NF-kappaB antibody demonstrated nuclear translocation of NF-kappaB within 5 min following NMDA treatment. NMDA treatment also enhanced the DNA binding activity of nuclear NF-kappaB in a Ca2+-dependent manner. Treatment with 3 mM aspirin blocked the NMDA-induced activation of JNK and NF-kappaB. Neuronal death following a brief exposure to 100 microM NMDA was Ca2+ dependent and attenuated by addition of aspirin or sodium salicylate. The present study suggests that Ca2+ influx is required for NMDA-induced activation of JNK and NF-kappaB as well as NMDA neurotoxicity. This study also implies that aspirin may exert its neuroprotective action against NMDA through blocking the NMDA-induced activation of NF-kappaB and JNK. PMID- 9751170 TI - Activation of excitatory amino acid receptors in the rat hippocampal slice increases intracellular Cl- and cell volume. AB - The effects of glutamatergic excitotoxins on intracellular Cl- were investigated in the CA1 pyramidal cell layer of the hippocampal slice. Hippocampal slices from rats (14-19 days old) were loaded with 6-methoxy-N-ethylquinolinium chloride (MEQ), a Cl(-)-sensitive fluorescent probe with a fluorescence intensity that correlates inversely with intracellular [Cl-]. Slices were exposed for at least 10 min at 26-28 degrees C to N-methyl-D-aspartate (NMDA; 100 microM) or alpha amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA; 50 microM). A UV laser scanning confocal microscope was used to measure changes in MEQ fluorescence within area CA1 pyramidal cell soma. Both glutamate receptor agonists produced a rapid decrease in MEQ fluorescence that persisted after washout following a 10 min exposure. The effects of NMDA and AMPA were prevented by the competitive antagonists 2-amino-5-phosphonopentanoic acid and 6,7-dinitroquinoxaline-2,3 dione, respectively. Neither tetrodotoxin nor picrotoxin prevented the effect of NMDA or AMPA, indicating the lack of involvement of presynaptic mechanisms. The effects of NMDA and AMPA on MEQ fluorescence were dependent on the levels of extracellular Cl-, but only NMDA responses were dependent on the levels of extracellular Na+. Removal of Ca2+ from the superfusion medium did not alter the effects of NMDA or AMPA on MEQ fluorescence. In addition, neither the Ca2+ ionophore ionomycin nor the L-type voltage-gated Ca2+ channel agonist (Bay K 8644) decreased MEQ fluorescence. The effects of NMDA and AMPA on cell (somal) volume were also assessed with the fluorescent probe calcein acetoxymethyl ester. Both NMDA and AMPA decreased calcein fluorescence (indicating an increased cell volume), but this was preceded by the decrease in MEQ fluorescence (equivalent to an intracellular accumulation of approximately 20 mM Cl-). Thus, excitotoxins may cause Cl- influx via an anion channel other than the GABA(A) receptor and/or reduce Cl- efflux mechanisms to produce cell swelling. Such anionic shifts may promote neuronal excitability and cell death following an excitotoxic insult to the hippocampal slice. PMID- 9751171 TI - Differential effects of two protein kinase C inhibitors, calphostin C and Go6976, on pineal cyclic nucleotide accumulation. AB - In rat pinealocytes, protein kinase C (PKC) is involved in the alpha1-adrenergic mediated potentiation of beta-adrenergic-stimulated cyclic nucleotide responses; however, the specific PKC isozyme(s) involved in the potentiation mechanism remain unknown. In the present study, we compared the effects of two PKC inhibitors, calphostin C, a specific inhibitor of PKC, and Go6976, a selective inhibitor of PKC alpha and PKC beta1, on the adrenergic-stimulated cyclic nucleotide accumulation in rat pinealocytes. Surprisingly, Go6976 was found to have an enhancing effect on basal cyclic GMP and isoproterenol-stimulated cyclic AMP and cyclic GMP accumulation, an effect not shared by calphostin C. Go6976 also increased the norepinephrine- and ionomycin-induced potentiation of isoproterenol-stimulated cyclic AMP and cyclic GMP accumulation, whereas the effect of calphostin C was inhibitory. The enhancing effect of Go6976 was abolished in the presence of isobutylmethylxanthine or zaprinast, but not rolipram, suggesting that this effect of Go6976 may be mediated through type V or the retinal type of phosphodiesterase. Based on these observations, we propose that some of the PKC isozyme(s) inhibited by calphostin C are involved in the potentiation of beta-adrenergic-stimulated cyclic nucleotide responses and that they act by enhancing synthesis. However, PKC isozymes inhibited by Go6976 appear to be basally active and tonically inhibit cyclic nucleotide accumulation through their stimulatory action on phosphodiesterase. PMID- 9751172 TI - Butyrylcholinesterase antisense transfection increases apoptosis in differentiating retinal reaggregates of the chick embryo. AB - To investigate the roles of the enzymes butyryl- and acetylcholinesterase (BChE and AChE) in retinal proliferation and differentiation, we use reaggregated spheres from retinal cells of the 6-day-old chick embryo, forming cellular and fibrous areas homologous to all layers of a normal retina. Recently, we could suppress BChE expression by transfecting these so-called retinospheroids during their proliferation period with a pSVK3 expression vector containing a 5' fragment of the rabbit BChE gene in antisense orientation. Along with morphological changes, proliferation was significantly decreased. Here, we have studied the effect of antisense BChE suppression during the differentiation period of retinospheroids. As BChE is suppressed, the differentiation of AChE positive cells is increased, whereas the immunoreactivities for red and green cone-specific opsins are strongly reduced. Concomitantly, the rate of apoptosis as determined by propidium iodide uptake, by increased CPP 32-like caspase expression, and by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling and DNA fragmentation assays is roughly doubled, predominantly at the expense of degenerating photoreceptor precursors. This is further strong evidence that the proliferation marker BChE regulates an intricate balance between cell proliferation, cell differentiation, and programmed cell death in this in vitro retinal system. PMID- 9751173 TI - S100 beta increases levels of beta-amyloid precursor protein and its encoding mRNA in rat neuronal cultures. AB - S100beta has been implicated in the formation of dystrophic neurites, overexpressing beta-amyloid precursor protein (betaAPP), in the beta-amyloid plaques of Alzheimer's disease. We assessed the effects of S100beta on cell viability of, neurite outgrowth from, and betaAPP expression by neurons in primary cultures from fetal rat cortex. S100beta (1-10 ng/ml) enhanced neuronal viability (as assessed by increased mitrochondrial activity and decreased lactic acid dehydrogenase release) and promoted neurite outgrowth. Higher levels of S100beta (100 ng/ml, but not 1 microg/ml) produced qualitatively similar, but less marked, effects. S100beta also induced increased neuronal expression of the microtubule-associated protein MAP2, an effect that is consistent with trophic effects of S100beta on neurite outgrowth. S100beta (10 and 100 ng/ml) induced graded increases in neuronal expression of betaAPP and of betaAPP mRNA. These results support our previous suggestion that excessive expression of S100beta by activated, plaque-associated astrocytes in Alzheimer's disease contributes to the appearance of dystrophic neurites overexpressing betaAPP in diffuse amyloid deposits, and thus to the conversion of these deposits into the diagnostic neuritic beta-amyloid plaques. PMID- 9751174 TI - Rilmenidine elevates cytosolic free calcium concentration in suspended cerebral astrocytes. AB - Rilmenidine, a ligand for imidazoline and alpha2-adrenergic receptors, is neuroprotective following focal cerebral ischemia. We investigated the effects of rilmenidine on cytosolic free Ca2+ concentration ([Ca2+]i) in rat astrocytes. Rilmenidine caused concentration-dependent elevation of [Ca2+]i, consisting of a transient increase (1-100 microM rilmenidine) or a transient increase followed by sustained elevation above basal levels (1-10 mM rilmenidine). A similar elevation in [Ca2+]i was induced by the imidazoline ligand cirazoline. The transient response to rilmenidine was observed in Ca2+-free medium, indicating that rilmenidine evokes release of Ca2+ from intracellular stores. However, the sustained elevation of Ca2+ was completely dependent on extracellular Ca2+, consistent with rilmenidine activating Ca2+ influx. Pretreatment with thapsigargin, an inhibitor of the endoplasmic reticulum Ca2+-ATPase, abolished the response to rilmenidine, confirming the involvement of intracellular stores and suggesting that rilmenidine and thapsigargin activate a common Ca2+ influx pathway. The alpha2-adrenergic antagonist rauwolscine attenuated the increase in [Ca2+]i induced by clonidine (a selective alpha2 agonist), but not the response to rilmenidine. These results indicate that rilmenidine stimulates both Ca2+ release from intracellular stores and Ca2+ influx by a mechanism independent of alpha2-adrenergic receptors. In vivo, rilmenidine may enhance uptake of Ca2+ from the extracellular fluid by astrocytes, a process that may contribute to the neuroprotective effects of this agent. PMID- 9751175 TI - Interleukin-1 beta and tumor necrosis factor-alpha suppress dexamethasone induction of glutamine synthetase in primary mouse astrocytes. AB - Astrocytes play a key role in the protection of neurons from excitotoxicity by taking up excess glutamate and converting it to glutamine via the enzyme glutamine synthetase. In a number of cell types, glucocorticoid hormones induce glutamine synthetase. Glucocorticoids also down-regulate many genes induced by proinflammatory cytokines. As the glucocorticoid receptor has been shown to interact with transcription factors that may also be activated by the proinflammatory cytokines interleukin-1beta (IL-1beta) and tumor necrosis factor alpha (TNF-alpha), we hypothesized that IL-1beta or TNF-alpha might oppose the induction of glutamine synthetase by dexamethasone. Primary mouse cortical astrocytes were treated with 10(-7) M dexamethasone and doses of IL-1beta or TNF alpha ranging from 0.02 to 5 ng/ml or 0.05 to 20 ng/ml, respectively. We found that both cytokines attenuated the dexamethasone induction of glutamine synthetase protein at 24 h and that the effect was dose-dependent. We also found that IL-1beta and TNF-alpha inhibited the induction of glutamine synthetase mRNA by dexamethasone, and that the induction of enzymatic activity was similarly prevented by IL-1beta. As glutamine synthetase can be induced by physiological levels of glucocorticoids, the release of proinflammatory cytokines following acute injury or in neurodegenerative disorders may hinder the ability of astrocytes to protect neurons from excitotoxicity. PMID- 9751176 TI - Glutamatergic regulation of basal and stimulus-activated dopamine release in the prefrontal cortex. AB - The present study was undertaken to determine whether basal and stimulus activated dopamine release in the prefrontal cortex (PFC) is regulated by glutamatergic afferents to the PFC or the ventral tegmental area (VTA), the primary source of dopamine neurons that innervate the rodent PFC. In awake rats, blockade of NMDA or alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) receptors in the VTA, or blockade of AMPA receptors in the PFC, profoundly reduced dopamine release in the PFC, suggesting that the basal output of dopamine neurons projecting to the PFC is under a tonic excitatory control of NMDA and AMPA receptors in the VTA, and AMPA receptors in the PFC. Consistent with previous reports, blockade of cortical NMDA receptors increased dopamine release, suggesting that NMDA receptors in the PFC exert a tonic inhibitory control on dopamine release. Blockade of NMDA or AMPA receptors in the VTA as well as blockade of AMPA receptors in the PFC reduced the dopaminergic response to mild handling, suggesting that activation of glutamate neurotransmission also regulates stimulus-induced increase of dopamine release in the PFC. In the context of brain disorders that may involve cortical dopamine dysfunction, the present findings suggest that abnormal basal or stimulus-activated dopamine neurotransmission in the PFC may be secondary to glutamatergic dysregulation. PMID- 9751177 TI - Long-term effects of portacaval anastomosis on the 5-hydroxytryptamine, histamine, and catecholamine neurotransmitter systems in rat brain. AB - Portacaval anastomosis (PCA) in the rat is used as a model for portal systemic encephalopathy. Changes in the serotonergic, histaminergic, and catecholaminergic neurotransmitter systems are often found shortly after PCA. We have examined the long-term effects of PCA on the aminergic systems in brains of male Wistar rats, which 8 months previously had been subjected to PCA. Precursors, amines, and metabolites were assayed by HPLC. Eight months after PCA, the catecholamine levels were unchanged in all brain regions. In contrast, tryptophan was evenly increased throughout the brain. The accumulation of 5-hydroxytryptophan after decarboxylase inhibition (NSD-1015; 100 mg/kg i.p.) and the endogenous levels of 5-hydroxyindoleacetic acid were significantly higher in PCA rats, particularly in the hypothalamus and midbrain, whereas 5-hydroxytryptamine concentrations were unchanged. Histamine levels were elevated throughout the brain with the greatest increase found in the hypothalamus and in the striatum. tele-Methylhistamine levels were significantly elevated in cortex and hypothalamus. We conclude that 8 months after PCA, catecholaminergic systems had reestablished their homeostasis, whereas serotonergic and histaminergic systems still show profound disturbances in their function. With histamine, this is reflected as an increase in the amounts of both transmitter and metabolite; serotonergic neurons respond by increasing only the level of the metabolite. PMID- 9751178 TI - Regulation of serotonin2A receptor expression by an antisense oligodeoxynucleotide. AB - The regulation of 5-HT2A receptor expression by an antisense oligodeoxynucleotide, complementary to the coding region of rat 5-HT2A receptor mRNA, was examined in a cortically derived cell line and in rat brain. Treatment of A1A1 variant cells, which express the 5-HT2A receptor coupled to the stimulation of phosphatidylinositol (PI) hydrolysis, with antisense oligodeoxynucleotide decreased the maximal stimulation of PI hydrolysis by the partial agonist quipazine and the number of 5-HT2A receptor sites as measured by the binding of 2-[125I]-iodolysergic acid diethylamide. Treatment of cells with random, sense, or mismatch oligodeoxynucleotide did not alter the stimulation of PI hydrolysis by quipazine or 5-HT2A receptor number. Intracerebroventricular infusion of antisense, but not mismatch, oligodeoxynucleotide for 8 days resulted in a significant increase in cortical 5-HT2A receptor density and an increase in headshake behavior induced by the 5-HT2 receptor agonist 1-(2,5-dimethoxy-4 iodophenyl)-2-aminopropane. The density of cortical 5-HT2A receptors was not altered by administration of antisense oligodeoxynucleotide for 1, 2, or 4 days. We hypothesize that in brain this antisense oligodeoxynucleotide relieved some form of translational suppression, resulting in an increase in 5-HT2A receptor expression. PMID- 9751179 TI - [3H]homoquinolinate binds to a subpopulation of NMDA receptors and to a novel binding site. AB - NMDA receptors mediate several important functions in the CNS; however, little is known about the pharmacology, biochemistry, and function of distinct NMDA receptor subtypes in brain tissue. To facilitate the study of native NMDA receptor subpopulations, we have determined the radioligand binding properties of [3H]homoquinolinate, a potential subtype-selective ligand. Using quantitative receptor autoradiography, NMDA-specific [3H]homoquinolinate binding selectively labeled brain regions expressing NR2B mRNA (layers I-III of cerebral cortex, striatum, hippocampus, and septum). NMDA-specific [3H]homoquinolinate binding was low in brain regions that express NR2C and NR2D mRNA (cerebellar granular cell layer, NR2C; glomerular layer of olfactory bulb, NR2C/NR2D; and midline thalamic nuclei, NR2D). In forebrain, the pattern of NMDA-specific [3H]homoquinolinate binding paralleled NR2B and not NR2A distribution. In addition to NMDA displaceable binding, there was a subpopulation of [3H]homoquinolinate binding sites in the forebrain, cerebellum, and choroid plexus that was not displaced by NMDA or L-glutamate. In contrast, we found that the derivative of homoquinolinate, 2-carboxy-3-carboxymethylquinoline, markedly inhibited the NMDA insensitive binding of [3H]homoquinolinate without inhibiting the NMDA-sensitive population. [3H]Homoquinolinate may be useful for selectively characterizing NR2B containing NMDA receptors in a preparation containing multiple receptor subtypes and for characterizing a novel binding site of unknown function. PMID- 9751180 TI - Opposing contributions of NR1 and NR2 to protein kinase C modulation of NMDA receptors. AB - N-Methyl-D-aspartate (NMDA) receptors mediate increases in intracellular calcium that can be modulated by protein kinase C (PKC). As PKC modulation of NMDA receptors in neurons is complex, we studied the effects of PKC activation on recombinant NMDA receptor-mediated calcium rises in a nonneuronal mammalian cell line, human embryonic kidney 293 (HEK-293). Phorbol 12-myristate 13-acetate (PMA) pretreatment of HEK-293 cells enhanced or suppressed NMDA receptor-mediated calcium rises based on the NMDA receptor subunit composition. NR2A or NR2B, in combination with NR1(011), conveyed enhancement whereas NR2C and NR2D conveyed suppression. The PKC inhibitor bisindolylmaleimide blocked each of these effects. The region on NR2A that conveyed enhancement localized to a discrete segment of the C terminus distal to the portion of NR2C that is homologous to NR2A. Calcium 45 accumulation, but not intracellular calcium store depletion, matched PMA effects on NMDA receptor-mediated calcium changes, suggesting that these effects were not due to effects on intracellular calcium stores. The suppression of intracellular calcium transients seen with NR2C was eliminated when combined with NR1 splice variants lacking C-terminal cassette 1. Thus, the intracellular calcium effects of PMA were distinguishable based on both the NR1 splice variant and the NR2 subunit type that were expressed. Such differential effects resemble the diversity of PKC effects on NMDA receptors in neurons. PMID- 9751181 TI - Hypoxia enhances [3H]noradrenaline release evoked by nicotinic receptor activation from the human neuroblastoma SH-SY5Y. AB - We have used the human sympathetic neuronal line SH-SY5Y to investigate the effects of hypoxia on noradrenaline (NA) release evoked by either raised [K+]o (100 mM) or the nicotinic acetylcholine receptor (nAChR) agonist dimethylphenylpiperazinium iodide (DMPP). NA release was monitored by loading cells with [3H]NA and collecting effluent fractions from perfused cells kept in a sealed perifusion chamber. Cells were challenged twice with either stimulus and release was expressed as that evoked by the second challenge as a fraction of that evoked by the first. K+-evoked release was unaffected by hypoxia (PO2 approximately 30-38 mm Hg), but release evoked by DMPP was significantly increased. For both stimuli, replacement of Ca2+o with 1 mM EGTA abolished NA release. K+-evoked release was also dramatically reduced in the presence of 200 microM Cd2+ to block voltage-gated Ca2+ channels, but DMPP-evoked release was less affected. In hypoxia, DMPP-evoked Cd2+-resistant NA release was dramatically increased. Our findings indicate that hypoxia increases NA release evoked from SH SY5Y cells in response to nAChR activation by increasing Ca2+ influx through the nAChR pore, or by activating an unidentified Cd2+-resistant Ca2+-influx pathway. As acetylcholine is the endogenous transmitter at sympathetic ganglia, these findings may have important implications for sympathetic activity under hypoxic conditions. PMID- 9751182 TI - Calcium modulation of activation and desensitization of nicotinic receptors from mouse brain. AB - The effects of extracellular calcium on functional properties of nicotinic receptors from mouse thalamus were investigated. Previous studies have reported that calcium modulates the function of several neuronal nicotinic receptors. A 86Rb+ ion efflux assay was developed to measure nicotinic receptor function from brain tissue, and data indicate that alpha4beta2 receptors may mediate this response. Using the 86Rb+ efflux assay, calcium effects on receptor activation, desensitization induced by high, activating and low, subactivating concentrations of agonist, and recovery from desensitization were examined. Effects of calcium on the kinetics of ligand binding were also investigated. Calcium modulated receptor activation by increasing the maximal response to nicotine in a concentration-dependent manner, without affecting the EC50 of nicotine. Barium, but not magnesium, mimicked the effects of calcium on receptor activation. The increase in receptor activation could not be explained by changes in the ratio of activatable to desensitized receptors as assessed by the kinetics of ligand binding. Desensitization following activation was unaffected by calcium. Calcium, barium, and magnesium, however, increased the potency of nicotine for desensitization induced by exposure to low, subactivating concentrations of nicotine. Recovery from desensitization was not modulated by calcium. These data suggest that calcium modulates various functional aspects of nicotinic receptors from mouse brain and may do so via different mechanisms. PMID- 9751183 TI - Mechanisms for facilitation of nitric oxide-evoked [3H]GABA release by removal of hydroxyl radical. AB - We have investigated the mechanisms for enhancement of nitric oxide (NO)-evoked gamma-[3H]aminobutyric acid ([3H]GABA) release from mouse cerebrocortical neurons by hydroxyl radical (.OH) scavengers. .OH scavengers, such as N,N' dimethylthiourea (DMTU), uric acid, and mannitol, dose-dependently facilitated NO evoked [3H]GABA release evoked by NO liberated from S-nitroso-N acetylpenicillamine. Ionomycin-evoked [3H]GABA release, which was significantly inhibited by hemoglobin and an NO synthase, N(G)-methyl-L-arginine, was also enhanced by DMTU. These results indicate that GABA release evoked by both endogenous and exogenous NO is facilitated by .OH scavengers. These enhancing actions of .OH scavengers were completely abolished by Ca2+ removal from incubation buffer and by an L-type voltage-dependent Ca2+ channel (VDCC) inhibitor, nifedipine, whereas each .OH scavenger showed no effects on [3H]GABA release in the absence of NO. Inhibitors for P/Q- and N-type VDCCs had no effects on the enhancement. NO-induced 45Ca2+ influx was also dose-dependently enhanced by .OH scavengers, although 45Ca2+ influx was not altered by .OH scavengers in the absence of NO. Nifedipine abolished this enhancement of the NO-induced 45Ca2+ influx by .OH scavengers. These results indicate that the removal of .OH by its scavengers facilitates the NO-evoked [3H]GABA release dependent on Ca2+ and that this enhancement is due to the increase in Ca2+ influx via L-type VDCCs. PMID- 9751184 TI - Quantification of the GABA shunt and the importance of the GABA shunt versus the 2-oxoglutarate dehydrogenase pathway in GABAergic neurons. AB - We investigated the activity of the cerebral GABA shunt relative to the overall cerebral tricarboxylic acid (TCA) cycle and the importance of the GABA shunt versus 2-oxoglutarate dehydrogenase for the conversion of 2-oxoglutarate into succinate in GABAergic neurons. Awake mice were dosed with [1-(13)C]glucose, and brain extracts were analyzed by 13C NMR spectroscopy. The percent enrichments of GABA C-2 and glutamate C-4 were the same: 5.0 +/- 1.6 and 5.1 +/- 0.2%, respectively (mean +/- SD). This, together with previous data, indicates that the flux through the GABA shunt relative to the overall cerebral TCA cycle flux equals the GABA/glutamate pool size ratio, which in the mouse is 17%. It has previously been shown that under the experimental conditions used in this study, the 13C labeling of aspartate from [1-(13)C]-glucose specifically reflects the metabolic activity of GABAergic neurons. In the present study, the reduction in the formation of [13C]aspartate during inhibition of the GABA shunt by gamma vinyl-GABA indicated that not more than half the flux from 2-oxoglutarate to succinate in GABAergic neurons goes via the GABA shunt. Therefore, because fluxes through the GABA shunt and 2-oxoglutarate dehydrogenase in GABAergic neurons are approximately the same, the TCA cycle activity of GABAergic neurons could account for one-third of the overall cerebral TCA cycle activity in the mouse. Treatment with gamma-vinyl-GABA, which increased GABA levels dramatically, caused changes in the 13C labeling of glutamate and glutamine, which indicated a reduction in the transfer of glutamate from neurons to glia, implying reduced glutamatergic neurotransmission. In the most severely affected animals these alterations were associated with convulsions. PMID- 9751185 TI - Stimulation of neuropeptide Y overflow in the rat paraventricular nucleus by corticotropin-releasing factor. AB - Neuropeptide Y (NPY) and corticotropin-releasing factor (CRF) are present at high concentrations in the hypothalamus where they mediate important endocrine and autonomic functions. Morphological and physiological studies have suggested an interaction between these peptides, and opposing actions of CRF and NPY have been reported on feeding and other behaviors. This study investigated the effect of CRF on NPY release in vivo, measured by push-pull techniques, in the anesthetized rat. Push-pull probes implanted into the paraventricular nucleus of the hypothalamus (PVN) were perfused with modified Ringer solution containing bovine serum albumin at 15 microl/min, and the perfusate was lyophilized prior to NPY radioimmunoassay. NPY overflow from the rat PVN was increased threefold by perfusion of a depolarizing concentration of potassium (50 mmol/L KCl). When CRF was administered into the PVN via the push-pull cannula at 1 or 5 microg/ml, dose dependent increases in NPY overflow of two- and fivefold were observed (p < 0.05). These increases were abolished by prior intracerebroventricular (i.c.v.) administration of the CRF antagonist [D-Phe12,Nle(21,38),C(alpha)MeLeu32]CRF (12 41) at 1 or 5 microg/microl, respectively. NPY overflow returned promptly to resting levels following CRF administration. In contrast, when CRF was administered by i.c.v. bolus at a similar total dose (2 microg), no significant effect on NPY overflow was observed. These data provide in vivo evidence for an interaction between CRF and NPY at the level of the PVN. PMID- 9751186 TI - Cannabinoid receptor activation differentially regulates the various adenylyl cyclase isozymes. AB - Two cannabinoid receptors belonging to the superfamily of G protein-coupled membrane receptors have been identified and cloned: the neuronal cannabinoid receptor (CB1) and the peripheral cannabinoid receptor (CB2). They have been shown to couple directly to the G(i/o) subclass of G proteins and to mediate inhibition of adenylyl cyclase upon binding of a cannabinoid agonist. In several cases, however, cannabinoids have been reported to stimulate adenylyl cyclase activity, although the mechanism by which they did so was unclear. With the cloning of nine adenylyl cyclase isozymes with various properties, including different sensitivities to alpha(s), alpha(i/o), and betagamma subunits, it became important to assess the signaling pattern mediated by each cannabinoid receptor via the different adenylyl cyclase isozymes. In this work, we present the results of cotransfection experiments between the two types of cannabinoid receptors and the nine adenylyl cyclase isoforms. We found that independently of the method used to stimulate specific adenylyl cyclase isozymes (e.g., ionomycin, forskolin, constitutively active alpha(s), thyroid-stimulating hormone receptor activation), activation of the cannabinoid receptors CB1 and CB2 inhibited the activity of adenylyl cyclase types I, V, VI, and VIII, whereas types II, IV, and VII were stimulated by cannabinoid receptor activation. The inhibition of adenylyl cyclase type III by cannabinoids was observed only when forskolin was used as stimulant. The activity of adenylyl cyclase type IX was inhibited only marginally by cannabinoids. PMID- 9751187 TI - Presenilin 1 mutations linked to familial Alzheimer's disease increase the intracellular levels of amyloid beta-protein 1-42 and its N-terminally truncated variant(s) which are generated at distinct sites. AB - Mutations in the presenilin genes PS1 and PS2 cause the most common form of early onset familial Alzheimer's disease. The influence of PS1 mutations on the generation of endogenous intracellular amyloid beta-protein (A beta) species was assessed using a highly sensitive immunoblotting technique with inducible mouse neuroblastoma (Neuro 2a) cell lines expressing the human wild-type (wt) or mutated PS1 (M146L or delta exon 10). The induction of mutated PS1 increased the intracellular levels of two distinct A beta species ending at residue 42 that were likely to be A beta1-42 and its N-terminally truncated variant(s) A beta x 42. The induction of mutated PS1 resulted in a higher level of intracellular A beta1-42 than of intracellular A beta x-42, whereas extracellular levels of A beta1-42 and A beta x-42 were increased proportionally. In addition, the intracellular generation of these A beta42 species in wt and mutated PS1-induced cells was completely blocked by brefeldin A, whereas it exhibited differential sensitivities to monensin: the increased accumulation of intracellular A beta x 42 versus inhibition of intracellular A beta1-42 generation. These data strongly suggest that A beta x-42 is generated in a proximal Golgi, whereas A beta1-42 is generated in a distal Golgi and/or a post-Golgi compartment. Thus, it appears that PS1 mutations enhance the degree of 42-specific gamma-secretase cleavage that occurs in the normal beta-amyloid precursor protein processing pathway (a) in the endoplasmic reticulum or the early Golgi apparatus prior to beta-secretase cleavage or (b) in the distinct sites where A beta x-42 and A beta1-42 are generated. PMID- 9751188 TI - Hypoxic cell death in human NT2-N neurons: involvement of NMDA and non-NMDA glutamate receptors. AB - Human NTera2 teratocarcinoma cells were differentiated into postmitotic NT2-N neurons and exposed to hypoxia for 6 h. The cultures were evaluated microscopically, and percent lactate dehydrogenase (LDH) release after 24 and 48 h was used as an assay for cell death. After 48 h LDH release was 24.3 +/- 5.6% versus 13.8 +/- 3.7% in controls (p < 0.001). Cell death was greatly diminished by MK-801 pretreatment (15.4 +/- 5.1%, p < 0.001). If glutamine was omitted from the medium, glutamate levels after 6 h of hypoxia were reduced from 101 +/- 63 to 2.3 +/- 0.3 microM, and cell death at 48 h was also markedly reduced (15.4 +/- 4.5%, p < 0.001). The alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (18.7 +/- 5.1%, p < 0.001) and mild hypothermia (33.5-34 degrees C) during hypoxia (19.5 +/- 2.7%, p < 0.05) were moderately protective. Basic fibroblast growth factor (24.1 +/- 3.2%), the nitric oxide synthase inhibitor N(G)-nitro-L-arginine methyl ester (22.8 +/- 8.1%), the antioxidant N-tert-butyl-o-phenyinitrone (18.9 +/- 5.9%), and the 21 aminosteroid U74389G (24.0 +/- 3.4%) did not protect the cells. N-Acetyl-L cysteine even tended to increase cell death (30.1 +/- 2.5%, p = 0.06). Treatment with MK-801 at the end of hypoxia did not reduce cell death (23.3 +/- 2.3%). In separate experiments, a 15-min exposure to 1 mM glutamate without hypoxia did not result in significant cell death (14.7 +/- 2.4 vs. 12.2 +/- 2.1%, p = 0.07). We conclude that, although somewhat resistant to glutamate toxicity when normoxic, NT2-N neurons die via an ionotropic glutamate receptor-mediated mechanism when exposed to hypoxia in the presence of glutamate. As far as we know, this is the first reported analysis of the mechanism of hypoxic cell death in cultured human neuronlike cells. PMID- 9751189 TI - Transcriptional and translational regulation of phosphodiesterase type IV isozymes in rat brain by electroconvulsive seizure and antidepressant drug treatment. AB - We examined the influence of electroconvulsive seizure (ECS) and imipramine (IMI) treatment on the transcription and translation of cyclic nucleotide phosphodiesterase type IV (PDE IV) isozymes in the rat brain. Our in situ hybridization studies revealed an increase of PDE IV-B mRNA level in various brain regions after acute ECS. However, the increase of PDE IV activity was produced not by acute but by chronic ECS treatment in the frontal cortex. Increased PDE IV-B mRNA expression in frontal but not in hippocampal subfields was induced also after chronic ECS treatment. Although an increase in PDE IV-A mRNA expression of the dentate gyrus in the hippocampus was observed, no change of PDE IV activity was produced in the hippocampus by acute or chronic ECS treatment. These results suggest that the repeated increases of PDE IV-B mRNA expression are attributable to the increase of PDE IV translation. Increased PDE IV-B transcription and PDE IV translation in the frontal cortex were also produced after chronic IMI treatment. This is the first report demonstrating an expressional regulation of Drosophila melanogaster dunce (dnc) gene homologue PDE IV isozymes in the brain. Although no pathophysiological conditions with reduced PDE IV activity in the nervous system are known except for a learning deficit in the mutant fly dnc-, our results suggest possible treatments to cope with reduced PDE IV activity. PMID- 9751190 TI - Proton-decoupled 31P magnetic resonance spectroscopy reveals osmotic and metabolic disturbances in human hepatic encephalopathy. AB - Quantitative proton and quantitative proton-decoupled 31P magnetic resonance spectroscopy (MRS) of the brain was performed in 16 patients with liver disease (10 with and six without chronic hepatic encephalopathy) and four patients with hyponatremia, as well as 20 age-matched normal subjects. Patients with hepatic encephalopathy were distinguished from controls by significant reduction in levels of cerebral nucleoside triphosphate (2.45 +/- 0.20 vs. 2.91 +/- 0.21 mmol/kg of brain; p < 0.0003), inorganic phosphate (p < 0.03), and phosphocreatine (p < 0.04). In addition of increased levels of cerebral glutamate plus glutamine and decreased concentrations of myo-inositol, patients with hepatic encephalopathy showed a reduction of total visible choline and of glycerophosphorylcholine (0.67 +/- 0.13 vs. 0.92 +/- 0.20 mmol/kg of brain in controls; p < 0.005) in 1H MRS, and of glycerophosphorylethanolamine (0.40 +/- 0.12 vs. 0.68 +/- 0.12 mmol/kg of brain in controls; p < 0.0003) in proton decoupled 31P MRS. Of the reduction of "total choline," 61% was accounted for by glycerophosphorylcholine, a cerebral osmolyte. Similar metabolic abnormalities were seen in hyponatremic patients. The results are consistent with disturbances of cerebral osmoregulation and energy metabolism in patients with chronic hepatic encephalopathy. PMID- 9751191 TI - Cellular localization and temporal elevation of tumor necrosis factor-alpha, interleukin-1 alpha, and transforming growth factor-beta 1 mRNA in hippocampal injury response induced by trimethyltin. AB - In certain pathologic states, cytokine production may become spatially and temporally dysregulated, leading to their inappropriate production and potentially detrimental consequences. Tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-1, IL-6, and transforming growth factor-beta (TGF-beta) mediate a range of host responses affecting multiple cell types. To study the role of cytokines in the early stages of brain injury, we examined alterations in the 17 day-old mouse hippocampus during trimethyltin-induced neurodegeneration characterized by neuronal necrosis, microglia activation in the dentate, and astrocyte reactivity throughout the hippocampus. By 24 h after dosing, elevations in mRNA levels for TNF-alpha, IL-1alpha, IL-1beta, and IL-6 mRNA were seen. TGF beta1 mRNA was elevated at 72 h. In situ hybridization showed that TNF-alpha and IL-1alpha were localized to the microglia, whereas TGF-beta1 was expressed predominantly in hippocampal pyramidal cells. Intercellular adhesion molecule-1, EB-22, Mac-1, and glial fibrillary acidic protein mRNA levels were elevated within the first 3 days of exposure in the absence of increased inducible nitric oxide synthetase and interferon-gamma mRNA. These data suggest that pro inflammatory cytokines contribute to the progression and pattern of neuronal degeneration in the hippocampus. PMID- 9751192 TI - Involvement of Bcl-2 family and caspase-3-like protease in NO-mediated neuronal apoptosis. AB - To clarify mechanisms of neuronal death in the postischemic brain, we examined whether astrocytes exposed to hypoxia/reoxygenation exert a neurotoxic effect, using a coculture system. Neurons cocultured with astrocytes subjected to hypoxia/reoxygenation underwent apoptotic cell death, the effect enhanced by a combination of interleukin-1beta with hypoxia. The synergistic neurotoxic activity of hypoxia and interleukin-1beta was dependent on de novo expression of inducible nitric oxide synthase (iNOS) and on nitric oxide (NO) production in astrocytes. Further analysis to determine the neurotoxic mechanism revealed decreased Bcl-2 and increased Bax expression together with caspase-3 activation in cortical neurons cocultured with NO-producing astrocytes. Inhibition of NO production in astrocytes by N(G)-monomethyl-L-arginine, an inhibitor of NOS, significantly inhibited neuronal death together with changes in Bcl-2 and Bax protein levels and in caspase-3-like activity. Moreover, treatment of neurons with a bax antisense oligonucleotide inhibited the caspase-3-like activation and neuronal death induced by an NO donor, sodium nitroprusside. These data suggest that NO produced by astrocytes after hypoxic insult induces apoptotic death of neurons through mechanisms involving the caspase-3 activation after down regulation of Bcl-2 and up-regulation of Bax protein levels. PMID- 9751193 TI - Role of carnitine palmitoyltransferase I in the control of ketogenesis in primary cultures of rat astrocytes. AB - The role of carnitine palmitoyltransferase I (CPT-I) in the control of ketogenesis was studied in primary cultures of rat astrocytes. Ketone bodies were the major product of [14C]palmitate oxidation by cultured astrocytes, whereas CO2 made a minor contribution to the total oxidation products. Using tetradecylglycidate as a specific, cell-permeable inhibitor of CPT-I, a flux control coefficient of 0.77 +/- 0.07 was calculated for CPT-I over the flux of [14C]palmitate to ketone bodies. CPT-I from astrocytes was sensitive to malonyl CoA (IC50 = 3.4 +/- 0.8 microM) and cross-reacted on western blots with an antibody raised against liver CPT-I. On the other hand, astrocytes expressed significant acetyl-CoA carboxylase (ACC) activity, and consequently they contained considerable amounts of malonyl-CoA. Western blot analysis of ACC isoforms showed that ACC in astrocytes--like in neurons, liver, and white adipose tissue--mostly comprised the 265-kDa isoform, whereas the 280-kDa isoform--which was highly expressed in skeletal muscle--showed much lower abundance. Forskolin was used as a tool to study the modulation of the ketogenic pathway in astrocytes. Thus, forskolin decreased in parallel ACC activity and intracellular malonyl-CoA levels, whereas it stimulated CPT-I activity and [14C]palmitate oxidation to both ketone bodies and CO2. Results show that in cultured astrocytes (a) CPT-I exerts a very high degree of control over ketogenesis from palmitate, (b) the ACC/malonyl-CoA/CPT-I system is similar to that of liver, and (c) the ACC/malonyl-CoA/CPT-I system is subject to regulation by cyclic AMP. PMID- 9751194 TI - Activation of JNK pathway and induction of apoptosis by manganese in PC12 cells. AB - Manganese is known to induce neurological disorders similar to parkinsonisms. A dopamine deficiency has been demonstrated in Parkinson's disease and in chronic manganese poisoning, suggesting that the mechanisms underlying the neurotoxic effects of the metal ion are related to a functional abnormality of the extrapyramidal system. However, the details have yet to be elucidated. Here we report that manganese causes characteristic internucleosomal DNA fragmentation, a biochemical hallmark of apoptosis, in PC12 cells. It was transcription dependent, relatively specific for manganese, and blocked in Bcl-2-overexpressed PC12 cells. The results indicate that apoptosis may play a role in the dopaminergic neurotoxicity associated with manganese, the first metal to be reported to induce this form of cell death. The early biochemical events show the impairment of energy metabolism, and the process may require new synthesis of proteins such as c-Fos and c-Jun. In addition, manganese induces phosphorylation of c-Jun at Ser63 and Ser73 and SEK1/MKK4 (c-Jun N-terminal kinase kinase) at Thr258 and tyrosine phosphorylation of several proteins. These results indicate that manganese activates specific signal cascades including the c-Jun N-terminal kinase pathway. PMID- 9751195 TI - In vivo aggregation of beta-amyloid peptide variants. AB - Transgenic Caenorhabditis elegans animals have been engineered to express wild type and single-amino acid variants of a long form of human beta-amyloid peptide (A beta 1-42). These animals express high levels (approximately 300 ng of A beta/mg of total protein) of apparently full-length peptide, as determined by quantitative immunoblot. Expression of wild-type A beta in these animals leads to rapid production of amyloid deposits reactive with Congo red and thioflavin S. This model system has been used to examine the effect of Leu17Pro, Leu17Val, Ala30Pro, Met35Cys, and Met35Leu substitutions on the in vivo production of amyloid deposits. We find that the Leu17Pro and Met35Cys substitutions completely block the formation of thioflavin S-reactive deposits, implicating these as key residues for in vivo amyloid formation. We have also constructed transgenic strains expressing a novel A beta variant, the single-chain dimer. Animals expressing high levels of this variant also fail to produce thioflavin S-reactive deposits. PMID- 9751196 TI - Apolipoprotein E attenuates beta-amyloid-induced astrocyte activation. AB - A common feature of Alzheimer's disease pathology is an abundance of activated glia, indicative of an inflammatory reaction in the brain. The relationship between glial activation and neurodegeneration is not known, although several cytokines and inflammatory mediators produced by activated glia have the potential to initiate or exacerbate the progression of neuropathology. As beta amyloid (A beta) is one of several stimuli that can activate glia, it is important to determine how A beta-induced glial activation is influenced by other proteins present in the plaque, such as apolipoprotein E (apoE). We examined the effect of native preparations of apoE on activation of rat cortical astrocyte cultures by A beta1-42. The apoE source was conditioned medium from human embryonic kidney 293 cells stably transfected with human apoE3 or apoE4 cDNA. By morphological criteria, apoE inhibited A beta-induced astrocyte activation in three experimental paradigms: apoE pretreatment blocked subsequent A beta-induced activation, A beta aged in the presence of apoE did not activate astrocytes, and apoE addition to activated astrocytes transiently reversed the activated phenotype. No apoE isoform selectivity was observed. The effect of apoE appears to be specific to A beta, as apoE did not attenuate cyclic AMP-induced astrocyte activation. These data suggest that apoE may modulate the ability of A beta to induce inflammatory responses in the brain. PMID- 9751197 TI - Aspirin and salicylate protect against MPTP-induced dopamine depletion in mice. AB - The neurotoxic effects of the dopamine-selective neurotoxin MPTP (15 mg/kg, s.c.), in mice, were totally prevented by systemic administration of salicylate (ED50 = 40 mg/kg, i.p.), aspirin (ED50 = 60 mg/kg, i.p.), or the soluble lysine salt of aspirin, Aspegic (ED50 = 80 mg/kg, i.p.). The protective effects of aspirin are unlikely to be related to cyclooxygenase inhibition as paracetamol (100 mg/kg, i.p.), diclofenac (100 mg/kg, i.p.), ibuprofen (20 mg/kg, i.p.) and indomethacin (100 mg/kg, i.p.) were ineffective. Dexamethasone (3-30 mg/kg, i.p.), which, like aspirin and salicylate, has been reported to inhibit the transcription factor NF-kappaB, was also ineffective. Aspirin or salicylate (100 microM) had no effect on dopamine uptake into striatal synaptosomes or on monoamine oxidase B activity. The neuroprotective effects of salicylate derivatives could perhaps be related to hydroxyl radical scavenging. This was suggested by the fact that hydroxylated metabolites of salicylate (2,3- and 2,5 dihydrobenzoic acid) were recovered in brain tissue following the combined administration of MPTP and aspirin to a greater extent than following aspirin alone. The surprising neuroprotective effects of aspirin in an animal model of Parkinson's disease warrant further clinical investigation. PMID- 9751198 TI - Increased levels of apolipoprotein D in cerebrospinal fluid and hippocampus of Alzheimer's patients. AB - Apolipoprotein D (apoD) is a member of the lipocalin family of proteins. Most members of this family are transporters of small hydrophobic ligands, although in the case of apoD, neither its physiological function(s) nor its putative ligand(s) have been unequivocally identified. In humans, apoD is expressed in several tissues, including the CNS, and its synthesis is greatly increased during regeneration of rat peripheral nerves. As apoD may have an important function in the nervous system and, particularly, in nerve regeneration, we measured immunoreactive apoD levels in the hippocampus and in CSF of patients with either Alzheimer's disease (AD) or other neuropathologies. In parallel, we determined the concentrations of apolipoprotein E (apoE), another apolipoprotein also implicated in nerve regeneration and in the etiology of AD. Levels of apoD but not apoE were increased in the hippocampus of AD patients compared with controls. ApoD concentrations, as determined by radioimmunoassay, were significantly increased in the CSF of AD patients (4.23 +/- 1.58 microg/ml) and patients with other pathologies (3.29 +/- 1.35 microg/ml) compared with those in the CSF of normal subjects (1.15 +/- 0.71 microg/ml). Although the differences were smaller than for apoD, the mean apoE concentrations in the CSF of both groups of patients were also significantly higher than those of controls. In AD patients, apoD, but not apoE, levels in CSF and hippocampus increased as a function of inheritance of the epsilon4 apoE allele. This study therefore demonstrates that increased apoD levels in the hippocampus and in CSF are a marker of neuropathology, including that associated with AD, and are independent of apoE concentrations. PMID- 9751199 TI - Putrescine-modified nerve growth factor: bioactivity, plasma pharmacokinetics, blood-brain/nerve barrier permeability, and nervous system biodistribution. AB - Previous investigations from our laboratory have demonstrated that the covalent modification of a variety of proteins, including antioxidant enzymes, with the naturally occurring polyamines--putrescine (PUT), spermidine, and spermine- dramatically increases their permeability coefficient-surface area product (PS) at the blood-brain and blood-nerve barriers after parenteral administration. In the present study, we have covalently modified nerve growth factor (NGF) with PUT by targeting carboxylic groups for their graded modification by controlling the ionization of these groups with pH. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis, western, and isoelectric focusing analyses demonstrated conversion of NGF to its polyamine-modified derivatives at different pH values. Although the immunoreactivity of PUT-NGF determined by ELISA and western analysis decreased with decreasing pH, the biological activity of PUT-NGF was not affected at any pH as determined by survival and neurite extension of dorsal root ganglia and PC12 cultures. Plasma pharmacokinetics after a single intravenous bolus administration revealed intact PUT-NGF through 10 min and 73-82% intact protein at 15 min. The PS value for PUT-NGF was maximized and the residual plasma volume (Vp) of the protein in the blood vessels minimized when the pH of the modification reaction was >6.4. The biodistribution of PUT-NGF at 15 min showed 22-33% intact protein in different brain regions, which represented 0.4-5.9 ng of PUT-NGF in different brain regions, a physiological dose that is capable of eliciting a bioresponse. The design of this polyamine-modified NGF derivative that has enhanced permeability at the blood-brain and blood-nerve barriers with retained bioactivity may obviate the necessity to create small-molecule mimics of NGF and may be applicable to neurotrophins, engineered multifunctional chimeric neurotrophins, antioxidant enzymes, and other therapeutic proteins with specific clinical application to neurological diseases. PMID- 9751200 TI - Depolarization of cerebellar granule cells increases phosphorylation of rabphilin 3A. AB - Studies performed over the past several years have provided evidence that phosphorylation of proteins is important in the regulation of neurotransmitter release. In this study, it is shown that rabphilin-3A is present in cerebellar granule cells as a phosphoprotein, by using 32P-labeling of cerebellar granule cells, immunoprecipitation, phosphoamino acid analysis, and phosphopeptide mapping. The level of phosphorylation was increased (224 +/- 13%) (mean +/- SEM) on depolarization of the cells with K+ (56 mM) in the presence of external Ca2+ (1 mM). Stimulation of protein kinase C with a phorbol ester (phorbol 12,13 dibutyrate) also enhanced the phosphorylation of rabphilin-3A (217 +/- 21%). Inhibitors of Ca2+/calmodulin-stimulated protein kinases or protein kinase C reduced the depolarization-enhanced phosphorylation of rabphilin-3A, indicating that rabphilin-3A is one of the targets for Ca2+-activated protein kinases in the nerve terminal. Costimulation of cells with phorbol 12,13-dibutyrate and K+ depolarization produced an increased level of phosphorylation of rabphilin-3A compared with either stimulus alone (287 +/- 61%). Phosphoamino acid analysis showed that serine was the main phosphorylated residue. A slight increase in the threonine phosphorylation could also be detected, whereas tyrosine phosphorylation could not be detected at all. These results suggest that rabphilin-3A is phosphorylated in vivo and undergoes synaptic activity-dependent phosphorylation during Ca2+-activated K+ depolarization. PMID- 9751201 TI - Role of serine-19 phosphorylation in regulating tyrosine hydroxylase studied with site- and phosphospecific antibodies and site-directed mutagenesis. AB - The effects of depolarization by elevated potassium concentrations were studied in PC12 cells and in stably transfected AtT-20 cells expressing wild-type or [Leu19]-recombinant tyrosine hydroxylase (rTH). Changes in the phosphorylation states of Ser19 and Ser40 in tyrosine hydroxylase (TH) were determined immunochemically using antibodies specific for the phosphorylated state of each site and compared with changes in TH activity in PC12 cell lysates and with changes in L-DOPA biosynthesis rates in intact AtT-20 cells. Treatment of either PC12 cells or AtT-20 cells expressing wild-type rTH with elevated potassium produced a transient increase in the phosphorylation state of Ser19 (up to 0.7 mol of phosphate/mol of subunit) in concert with a more gradual and sustained increase in Ser40 phosphorylation. Elevated potassium treatment also increased TH activity in PC12 cell lysates, but these increases paralleled the temporal course of Ser40, as opposed to Ser19, phosphorylation. Similarly, increases in DOPA accumulation produced by elevated potassium in AtT-20 cells expressing wild-type rTH paralleled the increases in the phosphorylation state of Ser40 but not Ser19. Moreover, elevated potassium produced comparable increases in DOPA accumulation in AtT-20 cells expressing rTH in which Ser19 phosphorylation had been eliminated (by substitution of Leu for Ser19). Thus, depolarization-induced increases in the stoichiometry of Ser19 phosphorylation do not appear to influence directly the activity of TH in situ. PMID- 9751202 TI - 7Li nuclear magnetic resonance study for the determination of Li+ properties in neuroblastoma SH-SY5Y cells. AB - Lithium has been used clinically in the treatment of manic depression. However, its pharmacologic mode of action remains unclear. Characteristics of Li+ interactions in red blood cells (RBCs) have been identified. We investigated Li+ interactions on human neuroblastoma SH-SY5Y cells by developing a novel 7Li NMR method that provided a clear estimation of the intra- and extracellular amounts of Li+ in the presence of the shift reagent thulium-1,4,7,10-tetrazacyclododecane N,N',N'',N'''-tetramethylene phosphonate (HTmDOTP4-). The first-order rate constants of Li+ influx and efflux for perfused, agarose-embedded SH-SY5Y cells in the presence of 3 mM HTmDOTP4- were 0.055 +/- 0.006 (n = 4) and -0.025 +/- 0.006 min(-1) (n = 3), respectively. Significant increases in the rate constants of Li+ influx and efflux in the presence of 0.05 mM veratridine indicated the presence of Na+ channel-mediated Li+ transport in SH-SY5Y cells. 7Li NMR relaxation measurements showed that Li+ is immobilized more in human neuroblastoma SH-SY5Y cells than in human RBCs. PMID- 9751204 TI - Characterization of granular particles isolated from postsynaptic densities. AB - We describe here the isolation and biochemical characterization of a population of protein aggregates from the postsynaptic density (PSD) prepared from pig cerebral cortex. The protein constituents of these aggregates are linked together primarily by disulfide bonds. Negative staining electron microscopy revealed that the isolated protein aggregates were granular objects with an average outside diameter of approximately 21 nm and with small protrusions on their surface. The major constituents of the isolated granular aggregates consist of tubulin and an unidentified protein of 70 kDa in size. Small amounts of the alpha subunit of calcium/calmodulin-dependent protein kinase II and subunits of alpha-amino-3 hydroxy-5-methyl-4-isoxazolepropionic acid and NMDA subtypes of glutamate receptors were also detected by immunoblotting. Actin, however, was not found in these granular aggregates. We propose that these granular protein aggregates correspond to the approximately 20-nm-diameter granular particles of the PSD on the basis of their biochemical and morphological characteristics. The spatial arrangement of these granular aggregates relative to other components of the postsynaptic terminal is also postulated here. PMID- 9751203 TI - Functional differentiation of multiple dopamine D1-like receptors by NNC 01-0012. AB - Although members of the multiple vertebrate/mammalian dopamine D1 receptor gene family can be selectively classified on the basis of their molecular/phylogenetic, structural, and tissue distribution profiles, no subtype specific discriminating agents have yet been identified that can functionally differentiate these receptors. To define distinct pharmacological/functional attributes of multiple D1-like receptors, we analyzed the ligand binding profiles, affinity, and functional activity of 12 novel NNC compounds at mammalian/vertebrate D1/D1A and D5/D1B, as well as vertebrate D1C/D1D, dopamine receptors transiently expressed in COS-7 cells. Of all the compounds tested, only NNC 01-0012 displayed preferential selectivity for vertebrate D1C receptors, inhibiting [3H]SCH-23390 binding with an estimated affinity (approximately 0.6 nM) 20-fold higher than either mammalian/vertebrate D1/D1A or D5/D1B receptors or the D1D receptor. Functionally, NNC 01-0012 is a potent antagonist at D1C receptors, inhibiting to basal levels dopamine (10 microM)-stimulated adenylyl cyclase activity. In contrast, NNC 01-0012 (10 microM) exhibits weak antagonist activity at D1A receptors, inhibiting only 60% of maximal cyclic AMP production by dopamine, while acting as a partial agonist at vertebrate D1B and D1D receptors, stimulating adenylyl cyclase activity by approximately 33% relative to the full agonist dopamine (10 microM), an effect that was blocked by the selective D1 receptor antagonist NNC 22-0010. These data clearly suggest that the benzazepine NNC 01-0012, despite lacking the N-methyl residue in the R3 position, is a selective and potent D1C receptor antagonist. Moreover, the differential signal transduction properties exhibited by NNC 01-0012 at these receptor subtypes provide further evidence, at least in vertebrates, for the classification of the D1C receptor as a distinct D1 receptor subtype. PMID- 9751205 TI - Vacuolar H+-ATPase domains are transported separately in axons and assemble in Torpedo nerve endings. AB - Torpedo electric organ synaptosomes possess a typical vacuolar H+-ATPase (V ATPase), inhibited by concanamycin A and insensitive to vanadate, made of the association of a catalytic soluble sector V1 to a membrane domain V0. In the electric nerves, the 57-kDa subunit B of the V1 sector was transported to the nerve endings by the slow axonal flow and did not accumulate upstream from an axonal block. In contrast, a 500% accumulation of the 15-kDa subunit c of the V0 membrane domain was observed, demonstrating that this subunit is conveyed by the fast axonal anterograde transport. After velocity sedimentation of solubilized nerve proteins, the 57- and 15-kDa subunits were recovered in different complexes corresponding, respectively, to the V1 and V0 domains. No fully assembled V ATPase was detected. It is concluded that V1 and V0 domains of V-ATPase are transported separately in axons, at different rates, and that they only associate once arrived in nerve endings to form the active V-ATPase. PMID- 9751207 TI - Regulation of oleoyl-CoA synthesis in the peripheral nervous system: demonstration of a link with myelin synthesis. AB - We studied the regulation of oleic acid synthesis in the PNS. During mouse postnatal development, the proportion of 18:1 rises in the sciatic nerve from 17% at 5 days of age to 33% at 25 days. However, this rise does not occur in the dysmyelinating mutant mouse trembler. In normal mouse development, the total stearoyl-CoA desaturase (SCD) activity measured in sciatic nerve homogenates is high during the first 3 weeks. Yet in trembler nerves, this SCD activity represents only 15% of normal values. Using the RT-PCR technique, we demonstrate that the SCD2 isoform is predominantly expressed in the PNS. Northern blot analysis showed that the mRNA levels for SCD2 parallel those of other specific myelin proteins in both normal mouse and trembler mutant development. Similar experiments in a rat demyelination-remyelination model confirmed that SCD2 mRNA levels are regulated in the PNS in a similar manner to myelin-specific proteins. PMID- 9751206 TI - Chronic imipramine administration amplifies the serotonin2A receptor-induced intracellular Ca2+ mobilization in C6 glioma cells through a calmodulin-dependent pathway. AB - In the present study, we examined whether chronic exposure of C6BU-1 cells to 100 nM of several different types of antidepressants directly influences serotonin2A (5-HT2A) receptor-stimulated intracellular Ca2+ mobilization. Imipramine, desipramine, clomipramine, and maprotiline amplified the 5-HT response at 48, but not at 2, h. Imipramine increased the maximum response to 5-HT without altering the EC50 of the dose-response curve. This effect was time dependent and cycloheximide blocked the maximal induction, suggesting an essential role for protein synthesis in this process. Previous exposure of the cells to thrombin or isoproterenol did not influence 5-HT2A receptor function and pretreatment with imipramine did not alter the thrombin- or bradykinin-induced Ca2+ mobilization, which indicates that the effects of imipramine appear to be specific to the 5 HT2A receptor. The effect of imipramine was potently suppressed by a calmodulin antagonist, W-13, in a dose-dependent manner. Furthermore, this amplified 5-HT response was blocked by KN-93, but not by H-7. Taken together, these results suggest that imipramine has a modulatory effect on the 5-HT2A receptor-coupled intracellular Ca2+ in C6 cells through a calmodulin-dependent pathway, possibly involving Ca2+/calmodulin kinase activation. PMID- 9751208 TI - Ca2+- and Mg2+-modulated lipolysis in neonatal rat brain slices observed by one- and two-dimensional NMR. AB - Superfused cortical brain slices from neonatal rats demonstrated large increases in levels of NMR-detectable lipids after sample preparation and perfusion with standard artificial CSF. These increases were reduced by an average of 58% by perfusion with buffer with low (no added) Ca2+ or by perfusion in Ca2+-free buffer. Perfusion with buffer with elevated MgSO4 (10 mmol/L) reduced the lipid changes by 47%. A reduction of 88% was observed in samples perfused in buffer with both low Ca2+ and high Mg2+, suggesting a role for Mg2+ in reducing lipolysis distinct from its known ability to block Ca2+ influx. PMID- 9751209 TI - Calcium/calmodulin-dependent protein kinase II is associated with NR2A/B subunits of NMDA receptor in postsynaptic densities. AB - NMDA receptors and Ca2+/calmodulin-dependent kinase II (CaMKII) have been reported to be highly concentrated in the postsynaptic density (PSD). Although the possibility that CaMKII in PSD might be associated with specific proteins has been put forward, the protein or proteins determining the targeting of the kinase in PSD have not yet been identified. Here we report that CaMKII binds to NR2A and NR2B subunits of NMDA receptors in PSD isolated from cortex and hippocampus. The association of NMDA receptor subunits and CaMKII was assessed by immunoprecipitating PSD proteins with antibodies specific for NR2A/B and CaMKII: CaMKII coprecipitated with NR2A/B and NR1 but not with other glutamate ionotropic receptor subunits, such as GluR1 and GluR2-3. A direct association between CaMKII and NR2A/B subunits was further confirmed by overlay experiments using either 32P autophosphorylated CaMKII or 32P-NR2A/B and by evaluating the formation of a CaMKII-NR2A/B complex by means of the cross-linker disuccimidyl suberate. These data demonstrate an association between the NMDA receptor complex and CaMKII in the postsynaptic compartment, suggesting that this colocalization may be relevant for synaptic plasticity. PMID- 9751210 TI - A molecular model of myelin oligodendrocyte glycoprotein. AB - Myelin oligodendrocyte glycoprotein (MOG) is a protein on the surface of myelin sheaths. It is a putative target of the autoimmune attack in the inflammatory and demyelinating CNS disease multiple sclerosis and its animal model, experimental autoimmune encephalomyelitis. MOG belongs to the immunoglobulin superfamily (IgSF), and its extracellular N-terminal domain contains many conserved IgSF consensus residues seen in immunoglobulin variable region folds. The aim of the present study was to create a molecular model of the extracellular N-terminal domain of mouse MOG. No crystal structure is yet available of MOG, and thus a molecular model would be useful in providing insight into its structure and binding characteristics. Molecular graphics techniques and molecular dynamics with secondary structure-based restraints were used in the construction and refinement of the MOG model. Regions of high prediction confidence were identified, and possible glycosylation, dimerization, complement binding, and antibody-binding regions in MOG were mapped and analyzed. PMID- 9751211 TI - In vitro evaluation of 11C-labeled (S)-nicotine, (S)-3-methyl-5-(1-methyl-2 pyrrolidinyl)isoxazole, and (R,S)-1-methyl-2-(3-pyridyl)azetidine as nicotinic receptor ligands for positron emission tomography studies. AB - The binding characteristics of the novel 11C-labeled nicotinic ligands (R,S)-1 methyl-2-(3-pyridyl) azetidine (MPA) and (S)-3-methyl-5-(1-methyl-2 pyrrolidinyl)isoxazole (ABT-418) were investigated in comparison with those of (S)-[11C]nicotine in vitro in the rat brain to be able to predict the binding properties of the new ligands for positron emission tomography studies in vivo. The data from time-resolved experiments for all ligands indicated fast binding kinetics, with the exception of a slower dissociation of [11C]MPA in comparison with (S)-[11C]nicotine and [11C]ABT-418. Saturation experiments revealed for all ligands two nicotinic receptor binding sites with affinity constants (K(D) values) of 2.4 and 560 nM and binding site densities (Bmax values) of 65.5 and 223 fmol/mg of protein for (S)-[11C]nicotine, K(D) values of 0.011 and 2.2 nM and Bmax values of 4.4 and 70.7 fmol/mg of protein for [11C]MPA, and K(D) values of 1.3 and 33.4 nM and Bmax values of 8.8 and 69.2 fmol/mg of protein for [11C]ABT 418. In competing with the 11C-ligands, epibatidine was most potent, followed by cytisine. A different rank order of potencies was found for (-)-nicotine, (+) nicotine, MPA, and ABT-418 displacing each of the 11C-ligands. Autoradiograms displayed a similar pattern of receptor binding for all ligands, whereby [11C]MPA showed the most distinct binding pattern and the lowest nonspecific binding. We conclude that the three 11C-labeled nicotinic ligands were suitable for characterizing nicotinic receptors in vitro. The very high affinity of [11C]MPA to nicotinic acetylcholine receptors, its low nonspecific binding, and especially the slower dissociation kinetics of the [11C] MPA from the putative high-affinity nicotinic acetylcholine receptor binding site compared with (S)-[11C]nicotine and [11C]ABT-418 raise the level of interest in [11C]MPAfor application in positron emission tomography. PMID- 9751212 TI - Up-regulation of blood-brain barrier short-form leptin receptor gene products in rats fed a high fat diet. AB - Leptin is a 16-kDa protein synthesized in adipose tissue that produces a satiety effect in the CNS. Leptin may gain access to the brain via receptor-mediated transport through the blood-brain barrier (BBB), and the BBB leptin receptor (OBR) may regulate the availability of circulating leptin to brain cells. The aim of the present study was twofold: first, to identify the OBR isoform expressed at the BBB, i.e., short, or "a," and long, or "b," form; and second, to compare the abundance of the BBB OBR mRNA and protein between control and high fat-fed rats. RT-PCR with isoform-specific primers showed that OBRa is the most abundant isoform at the BBB. BBB OBRa transcript content was markedly increased in high fat-fed rats compared with controls (11-fold), and no changes were observed in the expression of the internal standard control actin. The high fat feeding induction of OBR mRNA was correlated with an increase in the immunoreactive BBB OBR determined by immunocytochemistry using an all-isoform reactive antibody in high fat-fed obese rats. This investigation demonstrates (a) the OBRa is the principal leptin receptor expressed at the BBB and (b) this BBB OBR isoform is up regulated by a high fat diet. PMID- 9751214 TI - Alternative splicing at the 3'-cDNA of human tryptophan hydroxylase. AB - Two alternatively spliced transcripts of human tryptophan hydroxylase (TPH) were identified that differed at the 3' end of the open reading frame. Comparison of the human TPH cDNA and genomic sequences revealed that an intron containing an in frame stop codon could be alternatively spliced out of intron 11. This splicing would give rise to two human TPH isoforms with different C termini; the one that derives from the nonspliced intron contains a putative cyclic AMP-dependent protein kinase site, whereas the other one, which is 22 amino acids longer, does not. Analysis of various human tissues by RT-PCR revealed that the spliced TPH mRNA species was detected in all the postmortem tissues we tested, but the nonspliced species was expressed in only some tissues. PMID- 9751213 TI - Progesterone stimulates the activity of the promoters of peripheral myelin protein-22 and protein zero genes in Schwann cells. AB - To understand better the mechanisms by which progesterone (PROG) promotes myelination in the PNS, cultured rat Schwann cells were transiently transfected with reporter constructs in which luciferase expression was controlled by the promoter region of either the peripheral myelin protein-22 (PMP22) or the protein zero (P0) genes. PROG stimulated the P0 promoter and promoter 1, but not promoter 2, of PMP22. The effect of PROG was specific, as estradiol and testosterone only weakly activated promoters. Dose-response curves for stimulation of both promoter constructs by PROG were biphasic. RU486, a PROG antagonist, did not abolish the effect of PROG, but stimulated promoter activities by itself. In the human carcinoma cell line T47D expressing high levels of PROG receptor, PROG did not stimulate the P0 and PMP22 promoters, whereas the promoter region of the mouse mammary tumor virus was fully activated. Thus, the activation by PROG of promoter activity of two peripheral myelin protein genes is Schwann-cell specific. PMID- 9751215 TI - Persistent expression of Fas/FasL mRNA in the mouse hippocampus after a single NMDA injection. AB - Synaptic reorganization plays a very important role in brain adaptations to environmental stimuli, diseases, and aging processes. The NMDA model of excitotoxic injury was used to investigate the long-term molecular changes in the surviving neural cells in the mouse hippocampus. We demonstrated that a single intraperitoneal injection of NMDA produces persistent expression of c-fos, c-jun, Fas, and Fas ligand (FasL) mRNA in the hippocampus for 5 months. To determine the cellular origin of those gene transcripts in our in vivo model, a glial cell line and primary fetal neuronal culture were used to investigate the inducibility of the c-fos, c-jun, Fas, and FasL mRNA by NMDA. Both c-fos and Fas mRNA expression was observed in the NMDA-treated glial or neuronal cultures; however, c-jun and FasL mRNA was undetectable in this study. In our in vivo model, mossy fiber sprouting and apoptosis were also observed up to 40 days after the NMDA injection. Therefore, we hypothesize that the observed long-term expression of c fos, c-jun, Fas, and FasL mRNAs may reflect the ongoing synaptic reorganization. PMID- 9751216 TI - Acute effects of ethanol on pharmacologically isolated kainate receptors in cerebellar granule neurons: comparison with NMDA and AMPA receptors. AB - Comparisons of acute ethanol's effects on individual members of the three major families of ionotropic glutamate receptors (kainate, AMPA, and NMDA) have been performed only with recombinant receptors. However, no study has compared the acute effects of ethanol on individual members of each one of these receptor families in the same neuron. We accomplished this task by using cultured cerebellar granule neurons and LY303070 (GYKI-53784), a noncompetitive and selective AMPA receptor antagonist. Ethanol concentrations of 25, 50, 75, and 100 mM decreased the amplitude of pharmacologically isolated kainate-activated currents by 3 +/- 1, 9 +/- 2, 14 +/- 2, and 22 +/- 3% (n = 8), respectively. The magnitude of the ethanol-induced inhibition of nonselective kainate-activated currents, i.e., in the absence of LY303070, and currents activated by submaximal AMPA concentrations was not significantly different from that obtained with isolated kainate currents. However, the magnitude of the ethanol-induced inhibition of NMDA receptor-activated currents was about twofold greater than that of kainate and/or AMPA receptors. PMID- 9751217 TI - Insulin-like growth factors and bone: the osteoporosis connection revisited. AB - Tremendous advances have been made in knowledge about the pathogenesis and treatment of osteoporosis, a disease that affects more than 25 million Americans. In particular, it has been determined that two major processes are responsible for osteoporotic fractures. These are: 1) bone mass acquisition during adolescence; and 2) bone loss beyond the sixth decade. The former, and possibly the latter, are regulated by genetic and environmental factors. Insulin-like growth factor-I (IGF-I), a ubiquitous polypeptide, assumes a critical role in both of these processes. Very recent studies have elucidated a complex multifaceted IGF regulatory system in bone and have allowed investigators to consider site-directed approaches to therapy. Even more exciting is the prospect that the genetic regulation of peak bone mass may be controlled by components of the IGF regulatory system. Within the last half decade, tremendous strides have been made in defining the regulatory circuits that determine the expression of skeletal and serum IGF-I. These heritable modulators may be similar or identical to regulators of bone mineral density, thereby joining two distinct phenotypes. This minireview highlights some of the new investigations into the role IGF-I plays in the pathogenesis of osteoporosis. Although recent clinical trials with growth hormone and IGF-I in this disease have been relatively disappointing, advances on other fronts have generated considerable excitement, and these promise new and innovative approaches to this crippling disease. PMID- 9751218 TI - Leukemia cells and the cytokine network. AB - The cytokine network plays an important role in the growth and differentiation of normal and leukemic cells. Stimulation of this network, which has positive and negative regulators, results in the induction or inhibition of certain hematopoietic events. A cytokine can have multiple effects on various cell types, and combinations of cytokines with each other or with other exogenous substances produce more pronounced effects than any cytokine or agent individually. The mechanisms by which cytokines affect normal and leukemic cell growth and viability may vary depending on the target cell or the cytokine(s) in question. Diseases such as leukemia may result from abnormalities in the cytokine network or their receptors. Cytokines play a major role in leukemogenesis. Normally, hematopoietic cells require certain cytokines for their viability and growth. When the viability factors are withdrawn, apoptotic cell death naturally occurs. Prevention of programmed cell death by the abnormal production of a cytokine may release the cell from normal growth control leading to malignant transformation. Disregulation of genes for hematopoietic growth factors and their receptors may be one of the events that leads to leukemogenesis through an aberrant autocrine growth mechanism. However, cytokines have been used as therapeutic agents in various ways. Differentiation therapy has been widely investigated and proven effective in certain types of cancer. Gene therapy, where the cytokine cDNA is used to reduce tumorigenicity and/or increase immunogenicity is promising. Another kind of therapy using alkylated growth factors has been under focus. This review summarizes the actions and interactions of cytokines that are related to leukemic cell viability and growth. The use of cytokines as therapeutic agents is also discussed. PMID- 9751219 TI - Whole-egg diet delays the age-related impaired glucose tolerance of BHE/Cdb rats. AB - The effects of dietary egg on the age-related progression of impaired glucose tolerance and glomerulonephropathy in diabetes-prone BHE/Cdb rats were studied. This rat strain mimics the human with NIDDM. The development of impaired glucose tolerance was delayed in rats fed the whole-egg diet, however, feeding this diet resulted in elevated hepatic weight but had no effect on the age-related changes in renal lesions or renal function. We conclude that in this animal model for NIDDM, the development of glomerulonephropathy is independent of the development of impaired glucose tolerance and that diet can affect the time course for impaired glucose tolerance without affecting renal disease development. PMID- 9751220 TI - Cyclic AMP impairs the PRL stimulation of iodide uptake into mouse mammary tissues. AB - In earlier studies the uptake of iodide into mammary cells was found to occur by a mechanism similar to that in thyroid cells (i.e., the uptake occurs via a sodium-iodide symporter that is inhibited by perchlorate and thiocyanate). Although cyclic AMP stimulates the iodide transport mechanism in thyroid cells, the present studies show that cyclic AMP, as well as pharmacological agents that elevate cyclic AMP, impair iodide uptake into mammary cells. In addition, elevated cyclic AMP levels interfere with the PRL stimulation of iodide uptake as well as iodide incorporation into proteins in mammary tissues. Clearly, cyclic AMP has opposite effects on regulating iodide uptake processes in mammary versus thyroid cells. PMID- 9751221 TI - The influence of manganese deficiency on serum IGF-1 and IGF binding proteins in the male rat. AB - Young male rats subjected to a dietary manganese (Mn) deficiency respond to the deficiency by reducing their growth rate. The growth hormone (GH)/insulin-like growth factor (IGF) axis is critical for linear growth; this system is exquisitely sensitive to the nutritional state of the animal. In this study, we examined circulating GH, IGF-1, and insulin levels in Mn-deficient (-Mn; fed a 0.5 microg Mn/g diet) and sufficient (+Mn; fed a 45 microg Mn/g diet) male Sprague-Dawley rats. Additionally, we examined the distribution of circulating IGF binding proteins (IGFBPs) in animals of both dietary groups as these proteins modulate IGF-1 action in vivo and in vitro, and have been demonstrated to be altered in a number of nutritional and physiological states. Body weight was significantly reduced in -Mn relative to +Mn rats. Consistent with other studies, daily food intake was not altered. However, cumulative food intake (over 3 months) was marginally lower in -Mn versus +Mn animals. -Mn animals displayed lower circulating concentrations of IGF-1 (66% of control levels) and insulin (60% of control levels) despite having significant elevations in circulating GH levels relative to +Mn animals (140% of control levels). The IGFBP profile of -Mn animals reflected their elevated GH status, as we observed increased binding of tracer (125I-IGF-1) to the circulating IGFBP-3 complex (120% of control binding) using native chromatography techniques. Interestingly, the lower circulating insulin concentrations of -Mn animals did not result in dramatic elevations in lower-molecular-weight binding proteins. In summary, we demonstrate that in young male rats, Mn deficiency is associated with alterations in IGF metabolism. These alterations may contribute to the growth and bone abnormalities observed in -Mn animals. PMID- 9751222 TI - Metabolic abnormalities and differential responses to stress associated with hamster cardiomyopathy. AB - Metabolic differences between cardiomyopathic hamsters (CMHs), as they progress through various physiologic phases before reaching end-stage heart failure (HF), and healthy hamsters (HHs) are often difficult to demonstrate. We suggest that metabolic differences, magnified by application of chronic stress (S: cold immobilization 2 hr/day for 5 days) followed by acute stress (AS: 55 min global ischemia /30 min reperfusion), can be used to characterize different stages in this cardiomyopathic process. High performance liquid chromatography (HPLC) and 31P NMR methods were used to monitor the effects of acute stress applied to nonstressed (NS) and previously stressed CMHs (NS-2.5-month NS-5-month; S-2.5 month, S-5-month) and HHs (NS-HH, S-HH). Cardiac tissue extracts from nonstressed and stressed hamsters were analyzed for ATP and PCr at baseline and after completion of ischemia/reperfusion (AS) using HPLC. In nonstressed hamsters, ATP and PCr were 12% lower in CMHs (both NS-2.5- and NS-5-month) than in NS-HHs. After exposure to stress, ATP was 26% lower in CMHs (S-2.5- and S-5-month) compared to S-HHs, whereas there were minimal differences in PCr between the groups. 31P NMR monitoring of metabolism in the perfused beating heart during application of acute stress produced similar changes (%) in ATP and PCr in all groups (NS and S), whereas Pi increase was less in NS-5-month (118%) compared to NS-2.5-month (179%) and NS-HHs (306.8%), P < 0.05; and in S-5-month (148%) compared to S-2.5-month (216%) and S-HHs (222%). The changes in myocardial pH were inversely related to changes in Pi: NS-5-month (-13.5%); NS-2.5-month ( 9.7%); NS-HH (-17.7%). pH changes in stressed cardiomyopathic hamsters were similar to those of S-HHs. The postischemic recovery of ATP and Pi return closer to baseline values in cardiomyopathic hamsters (both NS and S) compared to healthy hamsters. The data suggest that cardiomyopathic hamsters have baseline metabolic abnormalities, and their responses to chronic cold immobilization stress, acute ischemia, and chronic cold immobilization stress plus acute ischemia are different from those in HHs. These responses may help to characterize specific stages of disease. PMID- 9751223 TI - Okadaic acid mimics several proximal effects of prolactin in Nb2 lymphoma cells. AB - We previously reported that prolactin-mediated macromolecular synthesis and mitogenesis are coupled to the activation of mitogen-activated protein kinase (MAPK) and p70 S6-kinase (p70S6K). Full activation of MAPK requires tyrosine and threonine phosphorylation whereas that of p70S6K requires serine phosphorylation. In the present study, okadaic acid, which inhibits serine/threonine protein phosphatase activity, was used to explore the linkage of MAPK and p70S6K activation to down-stream effects of prolactin in Nb2 cells. The results show that 1 nM okadaic acid augmented prolactin-stimulated mitogenesis and synthesis of protein and DNA 250%, 42%, and 70%, respectively. Addition of okadaic acid alone a) stimulated and sustained p70S6K activity (5- to 8-fold) and MAPK (3.5- to 5-fold); and b) increased protein synthesis with the maximum effect being about equal to that of prolactin (2.1-fold with 1 nMokadaic acid versus 2.3-fold with 0.2 nMprolactin). However, okadaic acid did not affect DNA synthesis or mitogenesis. These results indicate that the activation of MAPK and p70S6K is necessary for stimulation of protein synthesis but not sufficient for prolactin driven mitogenesis. PMID- 9751224 TI - Chelation of extracellular zinc inhibits proliferation in 3T3 cells independent of insulin-like growth factor-I receptor expression. AB - Depletion of zinc inhibits growth in animals and proliferation of cultured cells. Additionally, zinc can serve as an antioxidant protecting many compounds, including proteins, from oxidation. Regulation of cell division also involves insulin-like growth factor type I (IGF-I) and its receptor, especially during late G1 phase, allowing progression of the cell to S phase with subsequent DNA synthesis. We examined the effects of zinc depletion from the culture media of Swiss 3T3 cells on the cell cycle and IGF-I receptor expression. Cells were exposed to reduced fetal bovine serum concentrations to induce growth arrest, then returned to normal fetal bovine serum concentrations with the divalent cation chelator diethylenetriamine pentaacetic acid. Reducing the fetal bovine serum concentration did not induce quiescence in the cells as previously suggested. Zinc depletion reduced the proliferative fraction (S and G2/M phases) of the cell cycle. The addition of glutathione to the zinc-depleted media partially returned the proliferative fraction to the control level. Fetal bovine serum deprivation reduced IGF-I receptor expression whereas the absence of zinc had little effect on receptor expression. We conclude that depletion of zinc from culture media inhibits 3T3 cell proliferation independent of insulin-like growth factor-I receptor expression, and part of this inhibition is due to the antioxidant capacity of this divalent cation. PMID- 9751225 TI - Acute nicotine pretreatment augments dopaminergic pulmonary vasodilation. AB - Nicotine use has been associated with augmented dopaminergic neurotransmission within the central nervous system by a number of researchers. We wished to determine if acute nicotine treatment would augment the pulmonary vasodilatory response to dopamine. Male Sprague-Dawley rats were pretreated with either subcutaneous nicotine or equivolume saline and a dose-response curve for dopaminergic pulmonary vasodilation was constructed ex vivo in isolated, salt perfused rat lungs preconstricted with the synthetic thromboxane analogue U 46619. Nicotine pretreatment augmented the vasodilatory response to dopamine at doses falling within the mid range of the dopamine dose-response relationship, and this augmentation was blocked by the nicotinic ganglionic receptor antagonist mecamylamine. In contrast, nicotine did not augment the vasodilatory response produced by either the preferential DA1-dopaminergic receptor agonist SKF-38393 or the preferential DA2-dopaminergic receptor agonist quinpirole. Acute nicotine pretreatment did not affect the maximal pulmonary vasodilatory response produced by dopamine. Similarly, nicotine pretreatment failed to augment the vasodilatory response to the beta-adrenergic receptor agonists isoproterenol or terbutaline, but significantly diminished the response to dobutamine. Even though acute nicotine pretreatment did not augment beta-adrenergic receptor-mediated pulmonary vasodilation, the augmentation of dopaminergic responsiveness was inhibited by prior treatment with propranolol, suggesting beta-adrenergic receptor involvement. Finally, nicotine pretreatment did not alter the vasodilation produced by the nitric oxide dependent agents arginine vasopressin or sodium nitroprusside. These data demonstrate that nicotine alters dopaminergic vasodilation in the pulmonary vascular bed. PMID- 9751227 TI - Glucagon-like peptide-1 (7-36) amide administered into the third cerebroventricle inhibits water intake in rats. AB - Intracerebroventricular (icv) injection of glucagon-like peptide-1 (7-36) amide (GLP-1) has been shown to reduce food intake in rats. In these studies, we confirmed that injection of 10 microg of GLP-1 into the third cerebroventricle suppressed food intake. Moreover, we observed a reduction in water intake associated with the decreased food intake. We further examined whether GLP-1 injected icv in rats has a specific inhibitory effect on water intake. It was found that GLP-1 reduced water deprivation-induced drinking. Furthermore, the same dose of GLP-1 (10 microg) was sufficient to condition taste aversion. Finally, when 2 microg of GLP-1 were injected into the third ventricle, it only suppressed water deprivation-induced water intake and failed to influence spontaneous food and water intakes or induce conditioned taste aversion. These observations indicate that GLP-1 is a potent inhibitor of water intake in the rat and may play a role in the control of fluid homeostasis. PMID- 9751226 TI - Transient induction of polycystic ovary-like syndrome in immature hypothyroid rats. AB - Hypothyroidism in the human female is often associated with ovarian follicular cysts and hyperandrogenism, two cardinal signs of polycystic ovary syndrome. To explore the intraovarian changes that lead to follicular cyst formation in hypothyroidism, we have created a prepubertal hypothyroid rat model. These hypothyroid rats are hyperandrogenic and develop transient ovarian follicular cysts. Hypothyroidism in newborn rats was induced by providing the lactating dams with 0.04% propylthiouracil (PTU)-containing water. Subsequently, female rats were weaned and kept on PTU-containing water. On Day 25 of age, the rats were primed with 15 international units of pregnant mare's serum gonadotropin (PMSG) in 100 microl of phosphate buffered saline. Two days later, to initiate pseudopregnancy, they were injected with five international units of human chorionic gonadotropin (hCG). The animals were sacrificed at appropriate times, and blood and ovaries were collected for analyses. Control experiments were done with euthyroid rats. Two days after PMSG injection, well-developed antral follicles were observed in both the hypothyroid and euthyroid rats. Two days after hCG injection, while the euthyroid rat ovaries, as expected, contained numerous corpora lutea (CL), the hypothyroid rat ovaries still retained antral follicles. Some of these follicles with degenerating oocytes showed signs of luteinization. By 3-4 days post-hCG injection, the hypothyroid rat ovaries developed cystic follicles. By Day 6, however, the hypothyroid rat ovaries were indistinguishable from those of the euthyroid rats. Although serum testosterone concentrations were significantly elevated in the hypothyroid rats on Days 1-3, progesterone concentrations were not significantly different from the euthyroid animals. However, by Days 8-14, the hypothyroid rats had significantly higher serum progesterone concentrations. This model will be useful for investigating the intraovarian biochemical changes that lead to follicular cyst development in response to acute gonadotropin treatment. PMID- 9751229 TI - The relationship between smoking and triglyceride-rich lipoproteins is modulated by genetic variation in the glycoprotein IIIa gene. AB - In the last year, several studies have reported conflicting results concerning an association between the PI(A2) allele of the PI(A1/A2) polymorphism of platelet glycoprotein IIIa and the risk of myocardial infarction. In the present study, we analyzed the hypothesis of whether glycoprotein IIIa genotypes have any association with lipids and lipoproteins as classical cardiovascular risk factors. Smoking, associated with changes in triglyceride-rich lipoprotein (TRL) concentrations and with both hypercoagulability and reduced fibrinolysis, was also analyzed as an environmental factor. Blood samples were obtained from 170 subjects (83 men and 87 women; mean age, 57 years; SD 15) recruited by random sampling from the census of Girona, Spain. Subjects were classified as current smokers (n=41) and nonsmokers or exsmokers (n=129). Whereas no differences were found in lipid and lipoprotein concentrations between smokers and nonsmokers in subjects with the PI(A1/A1) genotype, smokers with the PI(A1/A2) or PI(A2/A2) genotypes showed significantly higher triglyceride and very-low-density lipoprotein (VLDL) triglyceride concentrations than nonsmokers or exsmokers with the same genotypes. Similarly, the VLDL triglyceride/HDL cholesterol ratio was significantly different in subjects with the PI(A1/A2) or PI(A2/A2) genotypes stratified according to smoking status. Further analysis revealed a significant interaction between smoking and genotype when those homozygous for the allele PI(A1) were compared with one or two PI(A2) alleles for the three lipidic parameters. The observed effects appear to show links between smoking, triglyceride metabolism, and a glycoprotein involved in platelet aggregation. It is likely that the pI(A) polymorphism is in linkage disequilibrium with other functional mutations that might influence triglyceride metabolism under some environmental factors such as smoking. This finding may provide a new perspective in the complex relationship between glycoprotein IIIa gene, environment, and their interactions. PMID- 9751228 TI - Insulin-induced vasodilation is dependent on tetrahydrobiopterin synthesis. AB - Insulin has been shown to elicit vasodilation through increases in nitric oxide (NO) production. To examine whether insulin may modulate the availability of tetrahydrobiopterin (BH4) (an absolute cofactor requirement for NO synthase activation), we studied the effects of insulin (150 nmol/L) on femoral arterial reactivity (to norepinephrine [NE]) in the presence and absence of 2,4-diamino-6 hydroxypyrimidine (DAHP), a specific inhibitor of BH4 production. Our data indicate that inhibition of BH4 synthesis results in an attenuation in the vasodepressor effect of insulin. One possibility is that insulin may regulate NO production by increasing cofactor (BH4) availability for activation of NO synthase. PMID- 9751230 TI - The effect of glucose and glucagon-like peptide-1 stimulation on insulin release in the perfused pancreas in a non-insulin-dependent diabetes mellitus animal model. AB - This study was designed to investigate the effect of glucogon-like peptide-1 (GLP 1) on pancreatic beta-cell function in normal, Zucker diabetic fatty (ZDF) rats, a model for non-insulin-dependent diabetes mellitus (NIDDM or type II diabetes) and their heterozygous siblings. Pancreas perfusion and enzyme-linked immunosorbent assay (ELISA) were used to detect the changes in insulin release under fasting and hyperglycemic conditions and following stimulation with GLP-1. Animals from the ZDF/Gmi-fa rats (ZDF) were grouped according to age, sex, and phenotype (obese or lean), and compared with LA lean rats. Glucose stimulation (10 mmol/L) in obese rats showed repressed response in insulin release. Glucose plus GLP-1 stimulation caused increased insulin release in all groups. The degree of this response differed between groups: lean > obese; young > adult; female > male. The LA lean control group was most sensitive, while the ZDF overtly diabetic group had the lowest response. In addition, the pulsatile pattern of insulin secretion was suppressed in ZDF rats, especially in obese groups. These results support the hypothesis that GLP-1 can effectively stimulate insulin secretion. Insulin release was defective in ZDF obese rats and could be partially restored with GLP-1. ZDF lean rats also showed suppression of beta-cell function and there was a difference in beta-cell function related to sex in ZDF strain. This study documents the efficacy of GLP-1 to stimulate insulin release and contributes to our understanding of the pathophysiological mechanisms underlying NIDDM. PMID- 9751231 TI - Role of female sex steroids in regulating cholesteryl ester transfer protein in transgenic mice. AB - The role of sex steroids in the regulation of cholesteryl ester transfer protein (CETP) was examined in the following groups of female transgenic mice carrying the human CETP gene: (1) normal, (2) ovariectomized, (3) ovariectomized and treated with estrogen; (4) ovariectomized and treated with progesterone; (5) ovariectomized and treated with both hormones, and (6) ovariectomized and treated with tamoxifen. CETP activity was measured in the plasma, and in the particulate and the soluble fractions of liver, muscle, and adipose tissue. Human CETP specific activity was determined by taking the difference of cholesterol ester transfer in the presence and absence of an antibody (TP2) against human CETP Ovariectomy reduced hormone levels, but did not completely abolish them from the circulation. Plasma CETP activity was significantly reduced in the tamoxifen group. There were significant reductions in CETP in liver homogenate and the soluble fraction, as well as in the particulate fraction of adipose with ovariectomy. Hormone replacement did not restore CETP activity in either the plasma or the tissues. Tamoxifin treatment resulted in a decrease in CETP activity in both fractions of liver, but had no effect on adipose. In the soluble fraction of adipose tissue and both fractions of muscle, only trace CETP activity was detected. We conclude that (1) minimal amounts of sex steroid hormones may be sufficient to affect CETP expression; (2) the effects of sex steroid hormones vary among tissues; and (3) in addition to the sex steroids, factor(s) from the ovary are needed for the full expression of CETP in this animal model. PMID- 9751232 TI - Amiodarone decreases gene expression of low-density lipoprotein receptor at both the mRNA and the protein level. AB - Amiodarone, a potent antiarrhythmic drug, decreases plasma and tissue triiodothyronine (T3) and increases plasma cholesterol levels, resembling changes seen during hypothyroidism. The increase of serum cholesterol during amiodarone medication is associated with a decreased expression of the hepatic low-density lipoprotein (LDL) receptor mRNA. To further elucidate the mechanism of amiodarone induced hypercholesterolemia, we investigated whether the decreased mRNA levels are the result of decreased transcription or increased degradation or both, and whether protein expression is decreased accordingly. Relative to pair-fed controls, amiodarone treatment increased plasma cholesterol by 69% and decreased expression of the mRNA encoding for the hepatic LDL receptor by 45%. To study this decrease in mRNA, we performed a run-on assay, from which it appears that amiodarone acts by decreasing LDL receptor mRNA expression 2.5-fold at the transcriptional level. The decay rate of liver LDL receptor mRNA, measured at different time points after injecting actinomycin D, was not different between amiodarone-treated and control animals (116+/-32 minutes and 84+/-10 minutes, P=.44). Hepatocytes in primary culture isolated from amiodarone-treated and control animals were used to determine specific binding of [125I]-LDL to hepatic LDL receptors. Amiodarone decreased specific LDL binding and Scatchard analysis demonstrated that amiodarone treatment reduced the number of LDL receptors by 69%, without affecting the dissociation constant (Kd). In conclusion, amiodarone induced hypercholesterolemia can be explained by decreased transcription of the LDL receptor gene, resulting in lower mRNA and protein levels. PMID- 9751233 TI - Therapeutic efficiency of lipoprotein(a) reduction by low-density lipoprotein immunoapheresis. AB - This study was performed to investigate the effect of low-density lipoprotein (LDL) immunoapheresis on lipoprotein(a) [Lp(a)] reduction in patients with heterozygous and homozygous familial hyperlipidemia (N=16) and insufficient response to lipid-lowering agents. By desorption of approximately 5,700+/-500 mL of plasma, a mean reduction in total cholesterol of 62% (P < .001) and in LDL cholesterol of 70% (P < .001) was achieved. Lp(a), which was elevated at study entry in seven of these patients (82.1+/-34.3 mg/dL; range, 48 to 148 mg/dL), was reduced during the initial LDL-apheresis procedure by 74.8%+/-14.1% (P < .001). Long-term apheresis treatment performed at weekly intervals resulted in an mean reduction in Lp(a) pretreatment values to 39.1+/-28.5 mg/dL (-54%; P < .001). Desorbed Lp(a) was measured at the waste of the columns for 31 apheresis treatments. Lp(a) concentration of the column waste was higher in patients with elevated serum Lp(a) pretreatment values as compared with those with Lp(a) serum values within the normal range (elevated Lp(a), 1,420+/-380 mg; without elevated Lp(a), 235+/-190 mg; P < .001). The rate of return of Lp(a) following apheresis treatment scheduled at weekly intervals was comparable to that of LDL cholesterol. PMID- 9751234 TI - Heparin inhibits human coronary artery smooth muscle cell migration. AB - Heparin, an anticoagulant, has been shown to reduce neointimal proliferation and restenosis following vascular injury in experimental studies, but the clinical trials of heparin in coronary balloon angioplasty have been negative. The current study, therefore, examined the effect of heparin on basal or stimulated migration by serum and platelet-derived growth factor (PDGF)-BB in cultured human coronary artery smooth muscle cells (SMCs) by Boyden's chamber method. In addition, the reversibility of the heparin effect on human coronary artery SMC migration was examined. Fetal calf serum (FCS) and PDGF-BB stimulated SMC migration in a concentration-dependent manner. Heparin in moderate to high concentration (10 to 100 U/mL) exhibited concentration-related inhibition of FCS- and PDGF-BB stimulated SMC migration; however, a low concentration (1 U/mL) of heparin had no inhibitory effects. Heparin also had weak inhibitory effects on nonstimulated SMC migration. The SMCs that were exposed to a high concentration (100 U/mL) of heparin for 6 hours were capable of migrating after a short lag period of removal of heparin from the culture medium. These SMCs also showed recovery of responses to FCS and PDGF-BB by migrating significantly greater than the nonstimulated level. Furthermore, heparin-containing medium did not contain detached cells. These results indicate that heparin inhibits human coronary artery SMC migration, especially when stimulated by FCS or PDGF-BB, and that this inhibitory effect of heparin is reversible and not simply a function of killing cells. PMID- 9751235 TI - Specific and scavenger low-density lipoprotein receptors involved in the disturbed lipid metabolism of patients with non-insulin-dependent diabetes mellitus are independent of obesity. AB - Comparative studies were performed on monocyte-derived macrophages (MDMs), prepared by a 72-hour incubation of blood monocytes obtained from patients with non-insulin-dependent diabetes mellitus (NIDDM) and age-matched obese and non obese controls. The MDMs, after a 72-hour culturing, expressed both specific and scavenger low-density lipoprotein (LDL) receptors on their surfaces. To study the binding capacity of both receptor types, [125I]LDL and [125I] acetylated LDL (acLDL) were applied to cells and the labeled ligands were then monitored to estimate the rate of intracellular degradations. The LDL-induced inhibition of endogenous cholesterol synthesis and the acLDL-triggered apolipoprotein (apo) E secretion were also studied, as the biological marker of receptor activation. The results indicate that the binding capacities of both specific and scavenger LDL receptors were not reduced in MDMs of diabetic patients. However, the intracellular degradation after LDL incorporation was decreased. The LDL-induced inhibition of cholesterol synthesis and the acLDL-transmitted apo E secretion were also found to be decreased in the MDMs of patients with NIDDM as compared with the obese and non-obese control groups. The NIDDM-induced impaired signal transduction of both specific and scavenger LDL receptors suggests an unclarified functional alteration of both receptor structures. PMID- 9751236 TI - Improvements in blood pressure, glucose metabolism, and lipoprotein lipids after aerobic exercise plus weight loss in obese, hypertensive middle-aged men. AB - The clustering of metabolic abnormalities often associated with hypertension, including insulin resistance, glucose intolerance, and dyslipidemia, in middle aged men may be the result of a decrease in cardiovascular fitness (VO2max) and the accumulation of body fat with aging. This study examines the effects of a 6 month program of aerobic exercise training plus weight loss (AEX+WL) on VO2max, body composition, blood pressure (BP), glucose and insulin responses during an oral glucose tolerance test (OGTT), glucose infusion rates (GIR) during 3-dose hyperinsulinemic-euglycemic clamps at insulin infusion rates of 120, 600, and 3,000 pmol x m(-2) x min(-1), and plasma lipoprotein levels. Compared with eight non-obese, normotensive, sedentary men (age, 62+/-2 years; 19%+/-2% fat; BP, 117+/-4/72+/-2 mm Hg), the nine obese, hypersensitive, sedentary men studied (age, 56+/-1 year; 32%+/-1% body fat; BP, 147+/-3/93+/-2 mm Hg) initially had a larger waist girth and waist-to-hip ratio (WHR) and were more hyperinsulinemic and insulin resistant with lower GIR at the two lower insulin infusion rates of the clamp and had a 2.9-fold higher EC50, the insulin concentration producing a half-maximal increase in GIR. They had higher triglyceride (TG) and lower high density lipoprotein cholesterol (HDL-C) levels. The AEX+WL intervention reduced body weight by 9%, percent body fat by 21%, waist girth by 9%, and WHR by 3%, and increased VO2max by 16% (P < .01 for all). This was associated with decreases of 14+/-3 mm Hg in systolic and 10+/-2 mm Hg in diastolic BP, significant changes in GIR at the low (+42%) and intermediate (+39%) insulin infusion rates and EC50 ( 39%) and in glucose (-21%) and insulin (-51%) responses during OGTT (P < .02 for all). AEX+WL also lowered total cholesterol by 14% and TG by 34%, and raised HDL2 C levels twofold (P < .01 for all). Thus, a 6-month AEX+WL intervention substantially lowers BP and improves glucose and lipid metabolism in obese, sedentary, hypertensive men. This suggests that hypertension and the metabolic risk factors for cardiovascular disease associated with it can be ameliorated by AEX+WL in obese, sedentary, middle-aged men. PMID- 9751237 TI - Alterations in the enzyme activity and protein contents of protein disulfide isomerase in rat tissues during fasting and refeeding. AB - Protein disulfide isomerase (PDI) is an enzyme that participates in the formation of disulfide bonds. It is also known to be the subunits of some enzymes and the membrane-associated thyroid hormone-binding protein. In this study, we measured the quantitative distribution of PDI protein in rat tissues and examined the relationship between protein level and enzyme activity in PDI during fasting and refeeding. Western blotting with specific anti-PDI antiserum detected the PDI protein band of 55 kd. Among several tissues, liver contained the largest amount of PDI protein, followed by kidney and fat, in which one-third to one-fourth of the hepatic PDI protein existed. The PDI protein band was also detected in heart and muscle. Fasting for 3 days decreased PDI protein levels in rat liver by 40%; control levels were recovered after 3 days of refeeding. The same change was observed in kidney. PDI activity, measured by the scrambled ribonuclease method, did not show the parallel alteration to PDI protein level in liver and kidney. Isomerase activity decreased to 50% of control values during fasting, but did not recover by refeeding. Thyroidal status did not affect either PDI protein level or isomerase activity. These findings show that fasting and refeeding affect PDI protein and enzyme activity, and that PDI protein level does not always reflect PDI activity. PMID- 9751238 TI - Reversal of diet-induced obesity and diabetes in C57BL/6J mice. AB - We have previously shown that C57BL/6J (B6) mice develop severe obesity and diabetes if weaned onto high-fat diets, whereas A/J mice tend to be obesity and diabetes-resistant. The purpose of this study was to determine if obesity and diabetes in the B6 mouse could be completely reversed by reducing dietary fat content. After 4 months, both strains consumed more calories on a high-fat diet than on a low-fat diet, and both strains showed a higher feed efficiency (FE=weight gained/calories consumed) on the high-fat diet versus the low-fat diet. However, relative to A/J mice, B6 mice demonstrated a significantly higher FE on the high-fat diet. Hyperglycemia, hyperinsulinemia, and increased adiposity were apparent in B6 mice after 4 months on the high-fat diet regardless of whether the diet was begun at weaning or 4 months later. Correlational analyses showed that adiposity was strongly related to both insulin and glucose levels in B6 mice, but only moderately related to insulin levels in A/J mice. In obese B6 mice that were switched to a low-fat diet, obesity and diabetes were completely reversed. Adiposity, fasting glucose, and fasting insulin values in these mice were equivalent to those in B6 mice of the same age that had spent 8 months on the low-fat diet. In summary, our data show that in the B6 mouse the severity of diabetes is a direct function of obesity and diabetes is completely reversible by reducing dietary fat. PMID- 9751239 TI - Equivalent efficacy of a time-release form of niacin (Niaspan) given once-a-night versus plain niacin in the management of hyperlipidemia. AB - This study compared the efficacy and safety of a once-a-night, time-release niacin formulation, Niaspan (Kos Pharmaceuticals, Miami Lakes, FL), with plain niacin and placebo for the treatment of primary hypercholesterolemia. The study was conducted in nine academic lipid research clinics in a randomized, double blind design. Niaspan 1.5 g at bedtime was compared with plain niacin 1.5 g/d after 8 weeks and 3.0 g/d after 16 weeks in divided doses and with placebo. A total of 223 hypercholesterolemic adult men and women participated. Compared with placebo at 8 weeks, Niaspan versus plain niacin at 1.5 g/d showed comparable efficacy, comparably lowering total cholesterol (C) (8%/8%), triglycerides (16%/18%), low-density lipoprotein (LDL)-C (12%/12%), apolipoprotein (apo B) (12%/12%), apo E (9%/7%), and lipoprotein(a) [Lp(a)] (15%/11%), and raising high density lipoprotein (HDL)-C (20%/17%), HDL2-C (37%/33%), HDL3-C (17%/16%), and apo A-I (8%/6%) (P < or = .05 in all instances). After 16 weeks, the Niaspan effect on LDL-C and triglyceride was unchanged while the plain niacin effect approximately doubled. At equal doses of 1.5 g/d of Niapan versus plain niacin, respectively, AST increased 5.0% versus 4.8% (difference not significant [NS]), fasting plasma glucose increased 4.8% versus 4.5% (NS), and uric acid concentrations increased less, 6% versus 16% (P=.0001). Flushing events were more frequent with plain niacin versus Niaspan (1,905 v 576, P < .001). Flushing severity was slightly greater with Niaspan, but still well tolerated. In conclusion, Niaspan 1.5 g hour of sleep (hs) has comparable efficacy, a lower incidence of flushing, a lesser uric acid rise, and an equivalent hepatic enzyme effect than 500 mg thrice-daily plain niacin in hyperlipidemic subjects. Niaspan may be an equivalent or better alternative to plain niacin at moderate doses in the management of hyperlipidemia. PMID- 9751240 TI - Effects of energy and purines in the diet on proliferation, differentiation, and apoptosis in the small intestine of the pig. AB - The effects of the energy and purine content in the diet on mucosal cell mitosis, function, and apoptosis in the small intestine of pigs were investigated in two experiments. In experiment I, three groups of five pigs were first fed a commercial diet that contained 9.1 MJ metabolizable energy (ME) per kilogram dry matter (DM) and 16.4% crude protein. It was followed by the experimental diets for 5 days each starting with an energy deficit (5.8 MJ ME/kg DM; 7% crude protein) followed by a high-energy diet with low purine content (14.1 MJ ME/kg DM; 13.6% crude protein; 460 mg purines/kg), or alternatively an isocaloric high purine diet (2,160 mg purines/kg). During experimental periods, blood samples were drawn daily through catheters for insulin-like growth factor-I (IGF-I) determination. The animals were killed at the end of the corresponding feeding period and gut tissue samples were collected. In tissue samples, IGF-I and parameters for the characterization of mitosis (thymidine kinase [TK], proliferating-cell nuclear antigen [PCNA]) and differentiation (RNA content, alkaline phosphatase, sucrase) were measured. The degree of apoptosis was determined histologically. In experiment II, five pigs were fitted with simple T cannula at the distal jejunum. They were fed the three experimental diets consecutively for 7 days each and sucrase and alkaline phosphatase were measured in digesta (four samples daily). IGF-I in blood but not in tissue clearly responded to the energy content of the diet with a decrease during the deficit and an increase in the two high-energy groups. However, purines had no additional effect on IGF-I. TK, PCNA, and gut weight showed an energy effect on mitosis, which was paralleled by increased peripheral IGF-I. Purines led to a further increase of mitosis, but IGF-I and gut weight were not increased. The degree of mitosis was correlated with higher activities of sucrase and alkaline phosphatase and also with the number of apoptotic cells. The enzyme activity increased from the deficit to the high-energy group and was further elevated due to purines. The results from experiment II also confirm these effects of energy and purines, because the activities of the enzymes in digesta decreased during energy deficit, but increased due to energy and in addition to purines. PMID- 9751241 TI - Influence of body fatness on the coronary risk profile of physically active postmenopausal women. AB - We have shown previously that endurance-trained postmenopausal runners demonstrate more favorable coronary heart disease (CHD) risk factors compared with age-matched sedentary women. However, the runners exhibited higher levels of physical activity and lower levels of body fatness, both of which can influence CHD risk factors. To gain insight into the influence of body fatness per se, we studied 38 postmenopausal healthy women: 10 swimmers, 10 runners, and nine obese and nine leaner sedentary subjects matched for age, hormone replacement use, and years postmenopause. Swimmers and runners were further matched for exercise training volume (4.5+/-0.2 v 4.6+/-0.6 h/wk) and relative competitive performance (79%+/-5% v 77+/-3% of age-adjusted world record). Maximal oxygen consumption (VO2max) on the treadmill was lower (P < .01) in swimmers versus runners. Body mass (65.0+/-2.0 v 59.0+/-1.3 kg), percent body fat (29%+/-2% v 23%+/-2%), and waist circumference (79+/-3 v 71+/-1 cm) were greater (P < .01) in swimmers than in runners. There were no significant differences in total caloric intake or dietary composition between swimmers and runners. Insulin sensitivity (via Bergman's minimal model) and fasting plasma concentrations of total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), triglyceride (TG), glucose, and plasminogen activator inhibitor-1 (PAI-1) activity were not different between the groups. However, plasma high-density lipoprotein cholesterol (HDL-C), HDL2-C, HDL-C/TC, insulin, fibrinogen, fibrin D-dimer, PAI antigen, tissue plasminogen activator (t-PA) activity, and t-PA antigen levels all were less favorable (P < .05) in swimmers versus runners. Daytime, nighttime, and 24-hour systolic blood pressure (SBP) was 6 to 10 mm Hg higher in swimmers compared with runners, but resting blood pressure, 24-hour blood pressure load, and blood pressure variability were not significantly different. Stepwise regression showed that measures of body fatness were the primary independent determinants of most of the metabolic CHD risk factors. When analysis of covariance (ANCOVA) was performed with body fatness as a covariate, differences in CHD risk factors between swimmers and runners were abolished (P=.18 to .90). We conclude that among endurance-trained postmenopausal women matched for training volume and competitive eliteness, higher total and abdominal body fatness is, in general, associated with a less favorable metabolic CHD risk profile. Thus, high levels of habitual aerobic exercise do not appear to negate the deleterious effects of adiposity on the coronary risk profile of healthy middle-aged and older women. PMID- 9751242 TI - Five-hour fatty acid elevation increases muscle lipids and impairs glycogen synthesis in the rat. AB - Insulin-mediated muscle glycogen synthesis is impaired after several weeks of high-fat feeding in rats, but not by short-term (2-hour) nonesterified fatty acids (NEFA) elevation induced by intravenous triglyceride/heparin infusion (TG/H). We examined whether a longer TG/H infusion induces defective glycogen synthesis. Five-hour hyperinsulinemic (700 pmol/L) euglycemic clamps with either TG/H or saline infusion were performed. TG/H-infused rats developed insulin resistance, but only after 2 to 3 hours. Red gastrocnemius glycogen synthesis rate decreased by 50% (P < .01 v saline) associated with decreased glycogen synthase activity (GSa; assessed at several glucose-6-phosphate [G-6-P] levels; two-way ANOVA, P=.02) and increased muscle TG and total long-chain acyl coenzyme A (LCAC) content (twofold; P < .05 v saline). Thus a 3- to 5-hour NEFA elevation in the rat produced significant impairment of insulin-stimulated muscle glycogen synthesis, associated with muscle lipid accumulation. These effects were similar to those observed after several weeks of fat feeding. The 5-hour TG/H-infused rat is a useful model for studying lipid-induced muscle insulin resistance. PMID- 9751243 TI - Serum insulin but not leptin is associated with spontaneous and growth hormone (GH)-releasing hormone-stimulated GH secretion in normal volunteers with and without weight loss. AB - Weight loss in humans is associated with elevated hypothalamic-pituitary growth hormone (GH) secretion. This study evaluates the effects of weight loss on the hypothalamic-pituitary (GH-releasing hormone [GHRH]-GH) axis in 14 normal-weight (body mass index [BMI], 25+/-1 Kg/m2) subjects, of whom half had undergone a diet induced weight loss of 14%+/-2% (mean+/-SEM). Insulin-like growth factor-1 (IGF 1), insulin, oral glucose tolerance, leptin, and GH pulse patterns were determined in both groups after weight maintenance for 1 week. Of note, we tested the effects of recent weight loss (3 months) and not a recent dietary intake, since both groups ingested a normal calorie diet for 2 days in the Clinical Research Center (CRC) prestudy. Serum insulin (3.8+/-0.7 v 9.0+/-0.9 microU/mL, P < .01) and C-peptide (0.44+/-0.06 v 0.59+/-0.04 ng/mL, P < .05) were significantly lower in the weight loss group. Serum leptin was not different. Endogenous GH pulse height (11.9+/-4.8 v 1.3+/-0.1 microg/L, P < .05), area per GH pulse ([AUC] 57+/-28 v 6+/-1 microg/L, P < .05), and mean GH (3.91+/-0.76 v 0.85+/-0.16 microg/L, P < .01) were increased in the weight loss group. The serum insulin level was inversely associated with the mean GH concentration (r=-.678, P < .01) and GH pulse height (r=-.733, P < .01). In addition to spontaneous GH secretion, the GHRH-stimulated GH pulse height (41.8+/-18.1 v7.1+/-1.6 microg/L, P < .05) and AUC (161+/-35 v46+/-13 microg/L/min, P < .05) were also increased in the weight loss group. The insulin concentration was also inversely correlated with the GHRH-stimulated GH pulse height (r=-.718, P < .01). The leptin concentration was correlated with the BMI (r=.554, P < .05) and body fat (r=.744, P < .01), but not with GH secretion. In summary, even though these patients were on a normal calorie diet, a history of recent weight loss in young men and women of normal weight and health can be associated with a significant increase in spontaneous GH pulse height and GHRH-stimulated pulse height. Weight loss was also associated with a reduced serum insulin level. The observed increase in GH secretion may be secondary to the reduction in insulin or alterations of other factors acting at the site of the pituitary. PMID- 9751244 TI - Resting metabolic rate in healthy adults: relation to growth hormone status and leptin levels. AB - Studies in patients with acromegaly and growth hormone (GH) deficiency, and administration of GH in normal and obese subjects and in patients with GH deficiency, suggest that GH increases resting metabolic rate (RMR) independently of changes in body composition. To test whether endogenous GH status determines RMR, we studied 38 healthy adults (18 women and 18 men) in two age groups (young, 30+/-0 years (n=18); older, 51+/-1 years [n=18]) with indirect calorimetry, deconvolution analysis of 24-hour GH secretion, arginin stimulation test, insulin like growth factor-I (IGF-I) measurement, lean and fat tissue distribution (computed tomography [CT] and dual-energy x-ray absorptiometry), assessment of physical fitness (maximal oxygen consumption [VO2max]), thyroid status, and serum leptin levels. RMR was higher in men compared with women, whereas RMR per lean body mass (LBM) (kcal x 24 h(-1) x kg(-1)) was higher in women (30.0+/-0.5 v 33.0 2/3 0.8; P=.003). GH secretion was higher in women and in young people. Total body fat (TBF) was higher in women, whereas LBM and abdominal fat were higher in men. Older people had significantly more TBF and abdominal fat as compared with younger people. VO2max was higher in younger people. Leptin levels were higher in women and in older people. Thyroid status was narrowly within the normal range in all subjects. RMR was strongly correlated with LBM (r=.90, P < .001). RMR/LBM correlated strongly with TBF (r=.49, P < .01) and leptin (r=.56, P < .001), but not with GH status. By multiple regression analysis, sex and TBF were the strongest predictors of RMR/LBM. However, in the young subgroup, GH production rate was a positive determinant of RMR/LBM. In the male subgroup, leptin was a stronger predictor than TBF of RMR/LBM (P < .001). Neither age, physical fitness, nor thyroid status contributed independently to predict RMR/LBM. In conclusion, (1) LBM was the most important determinant of RMR; (2) RMR/LBM was higher in women and depended strongly on TBF; (3) GH status in healthy adults was only weakly associated with RMR; and (4) in men, serum leptin levels were a strong positive determinant of RMR. PMID- 9751245 TI - Physiological reduction in fasting plasma glucose concentration in the first trimester of normal pregnancy: the diabetes in early pregnancy study. AB - Previous studies indicate that fasting plasma glucose decreases during gestation, but the timing and extent are not consistent from study to study. We had an opportunity to examine this question in the normal pregnancy cohort of women studied in the Diabetes in Early Pregnancy Study. Subjects were monitored to identify pregnancy by human chorionic gonadotropin testing, enrolled within 21 days of conception, and screened to rule out gestational diabetes at the juncture of the second and third trimesters. All subjects were instructed to fast overnight for 10 to 12 hours. Three hundred sixty-one women were studied between 6 and 12 weeks of gestation. A median decrease in plasma glucose of 2 mg/dL was observed between weeks 6 and 10 (P=.007). In a smaller group of subjects evaluated through the third trimester, little further glucose reduction was observed. A reduction in glycosylated hemoglobin levels between 10 and 20 weeks (P=.002) followed the earlier reduction in first trimester glucose levels. Analysis by body mass index (BMI) showed a smaller first trimester reduction with increasing BMI, and none among severely obese women (BMI > 29.9 kg/m2). The decline in fasting plasma glucose in pregnancy begins early in the first trimester, well before fetal glucose requirements can contribute to the decline in the glucose level. Thereafter, plasma glucose levels decrease little. These results suggest that in the setting in which this study was performed (an overnight fast) maternal physiologic adjustments account for a reduction in plasma glucose early in the first trimester of pregnancy, and possibly even later in gestation as well. PMID- 9751246 TI - Dietary protein restriction alters glucose but not protein metabolism in non insulin-dependent diabetes mellitus. AB - We determined whether a customary diet high or low in protein (1) influences postabsorptive amino acid catabolism, nitrogen (N) balance, and hepatic glucose output (HGO) in normal subjects or patients with non-insulin-dependent diabetes mellitus (NIDDM) or (2) alters blood glucose levels in NIDDM. Eight normal young adults and five obese middle-aged persons with NIDDM consumed low-protein (0.8 g/kg lean body mass [LBM]) or high-protein (3.0 g/kg LBM) diets at maintenance energy for consecutive 7-day periods. Fasting and average blood glucose and N balance were measured daily. The level of dietary protein had no effect on the basal plasma leucine rate of appearance (Ra) or urinary 3-methylhistidine excretion in either subject group. Basal leucine oxidation (and by inference, whole-body amino acid catabolism) was reduced on the low-protein diet but basal HGO was not, and although exogenous glucose effectively suppressed HGO, it did not reduce leucine oxidation with either diet. After adaptation to the low protein diet, N balance in both the normal and NIDDM subjects was close to zero. The low-protein diet reduced the fasting and daily blood glucose of the diabetic subjects by approximately 2 mmol/L (P < .05). We conclude that physiologic variation in dietary protein does not affect basal whole-body protein turnover or HGO in either normal young adults or obese middle-aged NIDDM subjects. However, protein restriction to the level of the average daily requirement significantly reduces postabsorptive and average daily blood glucose concentrations in persons with NIDDM. PMID- 9751247 TI - The effect of pioglitazone on glucose metabolism and insulin uptake in the perfused liver and hindquarter of high-fructose-fed rats. AB - To investigate the effect of pioglitazone, a thiazolidinedione oral antidiabetic agent, on the glucose and insulin metabolism in insulin resistance, a perfusion study of the liver and hindquarter was performed in high-fructose-fed rats. Male Wistar albino rats were assigned randomly to one of the following diets for 2 weeks: (1) normal chow (control group), (2) a diet high in fructose (fructose group), or (3) a high-fructose diet plus pioglitazone (pioglitazone intake of approximately 10 mg/kg body weight; pioglitazone group). The elevated levels of plasma insulin, triglyceride, and free fatty acids (FFA) in the fructose group were normalized by pioglitazone administration. In the perfused liver, the glucagon-induced increment in the glucose output of the fructose (57.1+/-9.1 micromol/g liver/20 min) and pioglitazone (44.7+/-10.1 micromol/g liver/20 min) groups was significantly (P < .01) higher than that in the control group (27.6+/ 5.7 micromol/g liver/20 min). The level in the pioglitazone group was significantly (P < .05) lower than that in the fructose group. In the presence of 100 or 500 microU/mL insulin, the insulin-mediated decrement in the glucagon induced glucose output of the fructose group (29.8+/-7.8 or 38.9+/-9.3 micromol/g liver/20 min) was significantly (P < .05) lower than that in the control (45.8+/ 14.2 or 54.5+/-8.5 micromol/g liver/20 min) and pioglitazone (44.4+/-9.2 or 56.2+/-10.8 micromol/g liver/20 min) groups, respectively. In the perfused hindquarter, glucose uptake in the fructose group (8.2+/-2.0 micromol/g muscle/30 min) was significantly (P < .05) lower than that in the control (12.1+/-2.3 micromol/g muscle/30 min) and pioglitazone (11.8+/-3.1 micromol/g muscle/30 min) groups. In the presence of 100 or 500 microU/mL insulin, glucose uptake in the fructose group (12.0+/-5.2 or 17.4+/-3.0 micromol/g muscle/30 min) was significantly (P < .05) lower than that in the control (20.2+/-2.4 or 23.0+/-3.1 micromol/g muscle/30 min) and pioglitazone (17.8+/-2.4 or 20.7+/-2.0 micromol/g muscle/30 min) groups, respectively. Insulin uptake by the liver and hindquarter was not significantly different in the control, fructose, and pioglitazone groups. These results indicate that pioglitazone improves the increased glucagon induced hepatic glucose output and decreases insulin-induced muscular glucose uptake without altering insulin uptake in high-fructose-fed insulin-resistant rats. PMID- 9751248 TI - Uric acid in chronic heart failure: a measure of the anaerobic threshold. AB - The anaerobic threshold (AT) is a measure of the balance between aerobic and anaerobic cellular metabolism. Hyperuricemia occurs in conditions that involve an imbalance between cellular oxygen consumption and carbon dioxide production, such as chronic heart failure (CHF). We therefore hypothesized that in CHF, serum uric acid might be related to the AT. Patients with CHF (n=40, aged 58.7+/-1.9 years; New York Heart Association Class I-IV; maximal oxygen consumption [MVO2], 18.7+/ 01.1 mL/kg/min; left ventricular ejection fraction, 26%+/-2%) and 10 age-matched healthy controls underwent measurement of the serum uric acid level at rest and assessment of the AT. This was derived from MVO2 and the regression slope relating minute ventilation to carbon dioxide output (VE - VCO2) during a maximal treadmill exercise test. Compared with the healthy controls, patients with CHF had a lower AT (11.8+/-0.7 v 16.9+/-1.1 mL/kg/min, P < .001) and a higher serum uric acid concentration (493.8+/-22.4 v 308.7+/-21.5 micromol/L, P < .001). In univariate analyses of the CHF group, the AT correlated with serum uric acid (r= .56, P < .001; AT=19.93 - (0.016 x uric acid), R2=.31, P < .001) and plasma creatinine (r=-.43, P < .01), but not with the diuretic dose. In stepwise regression analyses of the CHF group, serum uric acid emerged as a predictor of the AT (standardized coefficient=-.56, P < .001), whereas the diuretic dose and plasma creatinine failed to enter into the final models (multiple R2=.31, P < .001). In conclusion, in CHF there is an inverse relationship between the AT and the resting serum uric acid concentration. This is consistent with the known links between uric acid production and the imbalance in aerobic/anaerobic metabolism that occur in CHF. These findings provide the basis for using the simple measurement of the serum uric acid level as a surrogate measure of the AT. PMID- 9751249 TI - Upregulated synthesis of both apolipoprotein A-I and apolipoprotein B in familial hyperalphalipoproteinemia and hyperbetalipoproteinemia. AB - A family was identified with vertical transmission through three generations with simultaneous increases of apolipoprotein A-I (apoA-I), apolipoprotein B (apoB), low-density lipoprotein (LDL)-cholesterol, and high-density lipoprotein (HDL) cholesterol, which we have designated familial hyperalphalipoproteinemia and hyperbetalipoproteinemia (HA/HBL). Affected patients develop xanthomas and coronary artery disease (CAD). HA/HBL apoA-I and LDL-apoB were isolated and characterized. The in vivo kinetics of radiolabeled apoA-I and LDL-apoB were evaluated in two HA/HBL probands and three controls. Structural and metabolic characterization showed normal apoA-I and LDL-apoB. The kinetics of metabolism of HA/HBL apoA-I in the HA/HBL subjects showed that elevated apoA-I levels were solely due to an increased synthesis rate (15.2 to 17.6 mg/kg/d v 11.1 to 11.4 mg/kg/d) with a normal apoA-I residence time in plasma (4.2 to 5.4 days v 5.1 to 5.3 days). The elevation of LDL-apoB levels resulted from both an increased synthetic rate (16.6 to 22.9 mg/kg/d v 12.3 to 13.8 mg/kg/d) and a prolonged residence time (3.3 to 3.8 days v 1.4 to 1.9 days). In addition, we evaluated another HA/HBL proband of an unrelated family with HA/HBL to confirm the kinetic data. LDL-receptor binding studies of HA/HBL fibroblasts showed normal binding, uptake, and degradation of LDL isolated from a normolipemic control. The serum concentration of the cholesterol ester transfer protein (CETP) was normal in the studied probands. An apoB 3500 and apoB 3531 mutant, respectively, was ruled out by polymerase chain reaction (PCR). In conclusion, the site of the molecular defect in HA/HBL subjects may be involved in the coordinate regulation of metabolism for both LDL and HDL. PMID- 9751250 TI - Induction of apoptosis in prostate cancer cell lines by the green tea component, (-)-epigallocatechin-3-gallate. AB - Green tea components exert many biological effects, including antitumor and cancer preventive activities. In the search for anticancer agents for prostate cancer the inhibitory effects of green tea components were tested on the prostate cancer cell lines LNCaP, PC-3 and DU145. (-)-Epigallocatechin-3-gallate (EGCG) proved to be the most potent catechin at inhibiting cell growth. The inhibition induced by EGCG was found to occur via apoptotic cell death as shown by changes in nuclear morphology and DNA fragmentation. Thus, we report the first evidence that EGCG is the active component in green tea and induces apoptosis in human prostate cancer cells. PMID- 9751251 TI - Inhibitory effect of pentachlorophenol on gap junctional intercellular communication in rat liver epithelial cells in vitro. AB - To understand the initiating/promoting actions of pentachlorophenol (PCP), a non mutagenic hepatocarcinogen, and its metabolite, tetrachlorohydroquinone (TCHQ), we investigated the effects of each chemical on gap junctional intercellular communication (GJIC) in rat liver epithelial cells (WB cells) by the scrape loading and dye transfer method. After treatment with PCP, the GJIC was initially inhibited at 4 h but was restored in 6-8 h, followed by a second phase of inhibition between 16 and 24 h. Both the first and second inhibitions were concentration-dependent and were restored by 2-4 h after removal of PCP. The phosphorylation state of connexin 43 (CX43) and its localization on the plasma membrane were unchanged up to 24 h after treatment; however, this was accompanied by a decrease in the CX43 protein level. No inhibitory effect was apparent on the GJIC of cells treated with TCHQ. These results suggest that PCP may play a critical role of promoting activity via non-mutagenic mechanisms. PMID- 9751252 TI - Protein turnover in skeletal muscle of tumour-bearing transgenic mice overexpressing the soluble TNF receptor-1. AB - The implantation of the Lewis lung carcinoma (a fast-growing mouse tumour that induces cachexia) to both wild-type and transgenic mice for the soluble TNF receptor type I protein (sTNF-R1) resulted in a considerable loss of carcass weight in both groups. However, while in the wild-type mice there was a loss of both fat and muscle, in the transgenic mice muscle waste was not affected to the same extent as in the wild-type group. Muscle waste in wild-type mice was accompanied by an increase in the fractional rate of protein degradation, while no changes were observed in protein synthesis. The result was a decreased rate of protein accumulation which accounted for the muscle weight loss observed as a result of the tumour burden. In contrast, transgenic mice did not have such low rates of protein accumulation after tumour implantation. The increase in protein degradation in the tumour-bearing transgenic mice was accompanied by a similar increase in protein synthesis which compensated for the loss of muscle protein by degradation. Both tumour-bearing groups showed an enhanced expression of ubiquitin and proteasome C8 subunit genes, all of them related to the activation of the ATP-dependent proteolytic system in skeletal muscle. It is suggested that TNF may, in part, be responsible for the loss of protein in skeletal muscle of tumour-bearing mice. PMID- 9751253 TI - Azoxymethane-induced colon tumors and aberrant crypt foci in mice of different genetic susceptibility. AB - Azoxymethane (AOM) is an organotropic colon carcinogen that is commonly used to induce colon tumors in rodents. Unlike its parent compound, 1,2-dimethylhydrazine (DMH), a tumor susceptibility phenotype in inbred mice with respect to AOM has not been established. Thus, this study was undertaken to determine whether genetic susceptibility extends to this carcinogen. SWR/J, A/J (both susceptible to DMH carcinogenesis) and AKR/J (resistant) mice were treated with 10 mg/kg AOM i.p. once a week for 8 weeks. Twenty-five weeks after the initial injection, tumor yield was determined. With a single exception, only SWR/J and A/J mice developed tumors, with a distribution that was limited to the distal colon (16.3+/-1.1 and 36.4+/-2.4. respectively). The formation of aberrant crypt foci (ACF), putative preneoplastic lesions, was also assessed in whole-mount colons using Methylene Blue staining. Consistent with tumor multiplicity, the total number of ACF was highest in A/J mice, followed by SWR/J mice. In addition, A/J mice had a significantly greater number of large ACF (five or more crypts per foci) than the other strains. Despite the absence of colon tumors, however, AKR/J mice did develop a significant number of ACF. This finding suggests that ACF in resistant mice are persistent but do not progress to tumors. PMID- 9751254 TI - The expression of platelet-derived endothelial cell growth factor in human bladder cancer. AB - Platelet-derived endothelial cell growth factor (PD-ECGF) is an angiogenic factor and in some studies PD-ECGF/dTdRPase expression levels in several kind of cancers were higher than in their surrounding normal tissues. In this study, we evaluated PD-ECGF/dTdRPase expression in bladder cancer by an immunohistological method and determined whether it correlated with tumor stage, grade and recurrence. PD ECGF/dTdRPase expression was correlated with tumor grade and stage. Furthermore, among the superficial tumors, PD-ECGF/dTdRPase expression was correlated with a recurrence-free rate and thus it might be a prognostic factor for the recurrence of superficial bladder cancer. PMID- 9751255 TI - Glutathione S-transferase T1 and M1 gene defects in ovarian carcinoma. AB - Glutathione S-transferases (GSTs) M1 and T1 are known to be polymorphic in humans. Both polymorphisms are due to gene deletions, which are responsible for the existence of null genotypes. The gene defect of GSTT1 has been reported to be associated with an increased risk of myelodysplastic syndromes, astrocytoma and meningioma. A lack of GSTM1 was associated with tobacco smoke-induced lung and bladder cancer. In this study we examined whether the GSTT1 and/or GSTM1 homozygous null genotypes were associated with an increased risk of ovarian cancer using a multiplex polymerase chain reaction protocol. The GSTT1 null genotype was observed in 14% of the control subjects that had never suffered from neoplastic disease (n = 115) and in 16% of the patients affected with ovarian cancer (n = 103, OR 0.87, 95% CI 0.39-1.92, P = 0.73). A lack of GSTM1 was observed in 38% of the control subjects and in 46% of the patients (OR 0.77, 95% CI 0.44-1.32). This difference was not significant (P = 0.34). Similarly, no significant differences were obtained if GSTT1 and/or GSTM1 null genotypes were analyzed in subgroups of control subjects and ovarian cancer patients between the ages of 20-40, 41-70 and 71-90 years and in individuals with a positive family history of neoplastic disease. GSTT1 and/or GSTM1 null genotypes were not significantly associated with the histologic type and grade or FIGO (International Federation of Gynecology and Obstetrics) stages of the ovarian carcinomas. In conclusion, GSTT1 and/or GSTM1 null genotypes are not markers for an increased risk of ovarian cancer. PMID- 9751256 TI - Cathepsin D in tissue and serum of patients with squamous cell carcinoma of the head and neck. AB - Aspartic proteinase cathepsin D (CD) is believed to be associated with proteolytic processes leading to local invasion and seeding of tumour cells. To estimate a potential prognostic value of cathepsin D in squamous cell carcinoma of the head and neck, its total concentration was measured immunoradiometrically (ELSA-CATH-D kit, CIS bio international) in cytosols of tumour and adjacent normal tissue samples from 111 patients; in 42/111 patients, the CD concentration was determined in serum samples obtained at diagnosis (serum no. 1) and after the therapy (serum no. 2) from each of these patients. Sera of 15 healthy volunteers served as controls. A significantly elevated concentration of CD was measured in tumour cytosols as compared to normal tissue cytosols (31.1 versus 12.6 pmol/mgp, P < 0.0001) and in cytosols of normal laryngeal tissue than of the oral cavity or pharynx (13.3 versus 11.2 pmol/mgp, P = 0.03). The higher CD tumour concentration correlated with the age of the patients (< or =60 versus >60 years, 28.8 versus 32.8 pmol/mgp, P = 0.045) and histopathological tumour grade (G1+2 versus G3, 32.6 versus 24.4 pmol/mgp, P = 0.02). In serum samples, a lower concentration of CD was measured in the control group than in the patients (3.6 versus 4.1 pmol/mls, P = 0.045) and in serum no. 1 than in serum no. 2 (4.1 versus 5.1 pmol/ mls. P = 0.05). The CD concentration in sera obtained at diagnosis was stage dependent (S(I-III) versus S(IV), 3.9 versus 4.7 pmol/ mls. P = 0.09); there was a trend towards lower CD concentrations with an increasing time delay in serum no. 2 sampling (Rs = -0.20, P = 0.21). No correlation was observed between cytosolic and serum concentrations of CD. We conclude that our results confirm a specific role of CD in the process of invasion and metastasis of squamous cell carcinoma of the head and neck, which might also be of prognostic value in this particular cancer type. PMID- 9751257 TI - Induction of apoptosis by the N-acetyl-galactosamine-specific toxic lectin from Viscum album L. is associated with a decrease of nuclear p53 and Bcl-2 proteins and induction of telomeric associations. AB - The ribosome-inhibiting proteins from Viscum album L., i.e. the mistletoe lectins (ML), were recognized to induce apoptosis in various tumour cell lines and human lymphocytes. However, several aspects of ML-induced cell death are unclear. We report that the galNAc-binding ML III incubated with human lymphocytes mediates a very effective death signal resulting in the binding of Annexin-V and expression of mitochondrial membrane proteins Apo2.7, but also in an influx of the DNA intercalating dye propidium iodide. The addition of the ribosome-inhibiting protein Volkensin also induced Apo2.7 molecules, while Momordin, lacking a carbohydrate-binding chain, did not enter the cell membrane and thus did not affect the cells. However, we observed ML III to preferentially affect CD8+ cells with a memory phenotype (CD62L(lo)) as compared to their CD8+ CD62L(hi) counterparts, CD4+ T cells and CD19+ B cells. Furthermore, ML III did not induce sister chromatid exchange-inducing DNA lesions but reduced the intensity of telomeric signals, increased the frequencies of telomeric associations and C anaphases and reduced nuclear Bcl-2 and p53 proteins. Whatever the exact mechanisms are, our results provide strong evidence that the ML III-mediated cytotoxicity involves distinct killing pathways, i.e. (1) primary cell death via an induction of apoptosis which may not be dependent on protein and/or RNA synthesis and may not involve p53 and Bcl-2 proteins and (2) a loss of telomeres resulting in chromosomal instability in the surviving cells which is incompatible with life. However, we cannot exclude the possibility that this effect is due to a decrease in nuclear p53 proteins. PMID- 9751258 TI - Influence of intratumoral basic fibroblast growth factor concentration on survival in ovarian cancer patients. AB - Since basic fibroblast growth factor (bFGF) is considered as a potent mitogen that stimulates the growth of ovarian cancer cells, we evaluated the role of bFGF as a prognostic marker in patients with epithelial ovarian cancer. bFGF was quantified from the tumor cytoplasm of 76 patients with FIGO stage I-III ovarian cancer by a human FGF basic immunoassay (R&D Systems). After a mean follow-up period of 42 months, 50 patients were found to be free of tumor while 26 patients had died of the disease. The median bFGF concentration was 352.9 pg/mg (range 27.4-26600 pg/mg). After dichotomization cytoplasmic expression of bFGF was found to be low in 44 tumors (< or =500 pg/mg) and high in 32 tumors (>500 pg/mg). The probability of overall survival was 38.8 and 58.5% in the low bFGF and high bFGF groups, respectively (log-rank P = 0.0066). In multivariate analysis, residual tumor after initial surgery and bFGF, but not histologic grade or stage of the disease, independently influenced the overall survival probability. Furthermore, tumors with high cytoplasmic expression of bFGF revealed a much greater stromal content. Therefore, we hypothesize that bFGF may induce a fibroblastic response which causes tumors with a high bFGF to be less aggressive than those with less stromal tissue. PMID- 9751260 TI - Interstitial and large chromosome 1p deletion occurs in localized and disseminated neuroblastomas and predicts an unfavourable outcome. AB - We studied chromosome 1p loss of heterozygosity (1p-LOH) in 53 neuroblastomas (NBs) using 15 (CA)n repeat loci, which covered a region of 90 cM. We also assessed chromosome 1p36 deletion by fluorescence in situ hybridization (FISH) on interphase nuclei. 1p-LOH was found in 19 (36%, 95% confidence interval (CI) 23 50%) NBs. We detected interstitial and large deletion in both localized and disseminated tumours and in one tumour of a patient at stage 4S. Allelic loss was frequently observed in 1p36 and 1p32 regions. In patients older than 1 year of age (53 versus 13%, P < 0.002) we detected significant chromosome 1p deletion and it was associated with MYCN amplification (P = 0.001). Overall survival (OS) analysis showed that 1p-LOH is predictive of a poor outcome (odds ratio 16.5, 95% CI 5.4-50.9%); therefore, 1p-LOH should be regarded as an additional tumour progression marker in neuroblastoma. PMID- 9751259 TI - Porphyrin metabolism in rats intaking chemical carcinogens. AB - This study was performed on rats receiving nitrosamine precursors as potent liver carcinogens in order to investigate and follow up the porphyrin metabolism during the intake of hepatocarcinogens. As clarified from our results, progressive increases in free erythrocyte porphyrins, erythrocyte protoporphyrins, haem content and hepatic total porphyrins were observed after 2 months of intaking these carcinogens and further increases were gradually observed in parallel with the continued intake of these chemicals, which was extended to 7 months. At the same time, such elevations were also observed in the activity of hepatic delta aminolevulinic acid (ALA)-synthetase and uroporphyrinogen-1-synthetase either in hepatic tissues or erythrocytes of these carcinogen-subjected rats. However, significant inhibition was found in the activity of erythrocyte ALA-dehydratase, which reached 40.5% of the control values after 7 months. Therefore, these observations demonstrated that the intake of hepatocarcinogens may influence the rate of hepatic porphyrin and haem biosynthesis. PMID- 9751261 TI - A study on p16, pRb, cdk4 and cyclinD1 expression in non-small cell lung cancers. AB - To observe the expression of p16, pRb, cdk4 and cyclinD1 in non-small cell lung cancers, 104 cases of resected lung cancers were collected, which included squamous cell carcinomas, adenocarcinomas and large cell carcinomas. Immunohistochemistry assay was carried out. The results showed that 67% of squamous cell carcinomas and 46% of adenocarcinomas expressed p16, 64% of squamous cell carcinomas and 85% of adenocarcinomas expressed pRb and 66% of cancers expressed p16 or pRb. About 70% of the tumors expressed cyclinD1. More than 90% of the tumors expressed cdk4 and there was an increased trend with decreasing differentiation of both squamous cell carcinomas and adenocarcinomas. Sixty-seven percent of the highly differentiated and 100% of the poorly differentiated squamous cell carcinomas expressed cdk4. The aberrant p16 and pRb gene product expression played a significant role in the development and histological subtype of lung cancers by conditioning the biological behavior of NSCLC. cdk4 was an important factor in histological differentiation. PMID- 9751262 TI - p53-independent dephosphorylation and cleavage of retinoblastoma protein during tamoxifen-induced apoptosis in human breast carcinoma cells. AB - We have investigated several molecular events that occur during the process of tamoxifen-induced apoptosis in human breast carcinoma cells. We show that the treatment of either MCF-7 (containing wild-type p53) or MDA-MB-231 cells (containing mutant p53) with tamoxifen resulted in apoptotic nuclear changes and an increase in the pre-G1 apoptotic population. This was accompanied by activation of the caspase enzymes, as evidenced by specific cleavage of poly(ADP ribose) polymerase and retinoblastoma (RB) protein. The RB protein was cleaved at both an interior and carboxyl terminus cleavage site. In addition, dephosphorylation of RB was found at an early stage of tamoxifen-induced apoptosis in both cell lines. However, neither induction of p53 in MCF-7 cells nor induction of p21 in either cell line was detected, suggesting that tamoxifen induced RB dephosphorylation and apoptosis are independent of the p53/p21 pathway. We also observed an increase in levels of the pro-apoptotic Bax protein, the inhibitory cytokine TGF-beta1 and the transcription factor c-Myc in tamoxifen treated MDA-MB-231 cells, suggesting the possible involvement of these proteins during apoptosis in this system. PMID- 9751263 TI - Expression of platelet-derived endothelial cell growth factor (PD-ECGF) and its mRNA in uterine endometrial cancers. AB - To determine the potential of growth, invasion and metastasis of uterine endometrial cancer cells associated with neovascularization, the expressions of platelet-derived endothelial cell growth factor (PD-ECGF) and its mRNA in uterine endometrial cancers and in normal uterine endometria as controls were determined and the relationship between their expressions and histological grades, grades of myometrial invasion and clinical stages of uterine endometrial cancers was analyzed. The levels of PD-ECGF were significantly higher in uterine endometrial cancers of well-differentiated grade (G1) with invasion to < or =1/2 myometrium (B) and of stage 1 than in those of moderately and poorly differentiated grades (G2 and G3, respectively) limited to endometrium (A) and with invasion to >1/2 myometrium (C) and of stages II and III/IV and in normal uterine endometria. There was no significant difference in the levels between uterine endometrial cancers of G2 and G3, A and C, or stages II and III/IV and normal uterine endometria. Therefore, the active availability of PD-ECGF might contribute to the acceleration of angiogenic activity in the early process of invasion of well differentiated uterine endometrial cancers. PMID- 9751264 TI - Androgen induction of urokinase gene expression in LNCaP cells is dependent on their interaction with the extracellular matrix. AB - Urokinase-type plasminogen activator (uPA) plays a central role in tissue remodeling and cell invasion. In the present study, we examined the expression of uPA in the prostate cancer cell lines LNCaP, DU-145 and PC-3. In contrast to DU 145 and PC-3, the androgen-responsive cell line LNCaP does not express uPA. However, seeding LNCaP cells on fibronectin-coated plates stimulated a low level of uPA expression which was further induced upon exposure of the cells to dihydrotestosterone (DHT). Concomitant with the expression of uPA, an androgen regulated expression of uPA receptor (uPAR) was induced. These results suggest that the interaction of LNCaP cells with the extracellular matrix plays a dominant role in the androgen control of uPA and uPAR gene expression. PMID- 9751265 TI - Stannous chloride and the glucoheptonic acid effect: study of a kit used in nuclear medicine. AB - Stannous dichloride is used as a reducing agent in the preparation of technetium 99m radiopharmaceuticals. We decided to evaluate the genotoxic potential of the tin (II)-glucoheptonate complex in the kit using a tester strain of Escherichia coli AB1157. Our results show that tin (II) chloride and the tin (II) glucoheptonate complex exert a genotoxic effect in this system. While the genotoxic effect disappeared when the glucoheptonate concentration was increased, the glucoheptonate did not protect the cultures from the damaging effects of hydrogen peroxide. The ability of glucoheptonate to protect cultures from tin (II)-induced damage can be explained on the basis of its metal chelating properties. PMID- 9751266 TI - Multidrug resistance genes (MRP) and MDR1 expression in small cell lung cancer xenografts: relationship with response to chemotherapy. AB - Intrinsic or acquired drug resistance is a major limiting factor of the effectiveness of chemotherapy. Increased expression of either the MRP gene or the MDR1 gene has been demonstrated to confer drug resistance in vitro. In this study, we examined MRP and MDR1 gene expression in a panel of 17 small cell lung cancers (SCLC) xenografted into nude mice from treated and untreated patients using an RT-PCR technique. For some of them, the outcome of the corresponding patients was known and we related MDR1/MRP expression with the xenograft response to C'CAV (cyclophosphamide, cisplatin, adriamycin and etoposide) combined chemotherapy. Fifteen (88%) of the 17 cases of SCLC were found to be positive for either MDR1 or MRP. MRP gene expression was present in 12 (71%) of 17 cases, whereas MDR1 gene expression was detected in eight (50%) of 16 cases. For six SCLC, the survival duration of patients differed, with three patients surviving for more than 30 months after therapy. Among these six turnours, five expressed MRP and/or MDR1. These six xenografts responded to the C'CAV treatment but a significant rate of cure was obtained in only three cases. No obvious relationship was observed between the response to this treatment and MRP or MDR1 expression. However, the remarkably high levels and frequency of MRP expression in some SCLC samples indicate that future developments in chemotherapy of this tumour type should anticipate that drugs which are substrates of MRP may be of limited effectiveness. PMID- 9751267 TI - Genistein in the control of breast cancer cell growth: insights into the mechanism of action in vitro. AB - Genistein significantly inhibited cell growth (IC50 around 10 microM) of MCF-7, MDAMB-231 and HBL-100 cell lines, but not of skin-derived fibroblasts and counteracted the growth-stimulatory effects exerted by estradiol and growth factors. It abolished the paracrine stimulation observed in MCF-7 cells in co culture with MDAMB-231 or fibroblasts. Genistein-treated cells accumulated in the S and G2/M phases of the cell cycle and underwent apoptosis. Genistein decreased tyrosine phosphorylation induced upon treatment with transforming growth factor alpha. Finally, genistein bound the estrogen receptor (ER) (relative affinity constant Kd = 4 nM), induced pS2 and cathepsin-D transcription and increased nuclear ER levels. PMID- 9751268 TI - p53 exon 7 mutations as a predictor of poor prognosis in patients with colorectal cancer. AB - We have studied 61 resected colorectal adenocarcinomas in order to investigate p53 mutations as a prognostic factor for this pathology. Mutations in exons 5-9 of the p53 gene were analyzed by the polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) technique followed by sequencing. Our data indicate that p53 exon 7 mutations were prevalent in the latest stages of colorectal carcinogenesis and patients bearing this alteration had the worst prognosis. Therefore, according to our results, mutations affecting exon 7 of the p53 gene could be considered as a useful marker of biological aggressiveness for colorectal cancer. PMID- 9751269 TI - Treatment with field bean protease inhibitor can effectively repress ethylnitrosourea (ENU)-induced neoplasms of the nervous system in Sprague-Dawley rats. AB - The ability of field bean protease inhibitor (FBPI) to inhibit ethylnitrosourea (ENU)-induced tumours of the nervous system of Sprague-Dawley rats was investigated. Groups of 1-day-old rats were injected intraperitoneally (i.p.) with neurocarcinogenic amounts of ENU and a few hours later, one group was treated i.p. with 80 mg of FBPI per kg body weight. This treatment was carried out three times a week for the first month and five times a week for the next month. Animals were killed when they were neurologically ill and their neural tissues were assessed for lesions. Those FBPI-treated rats which showed no illness were also killed to terminate the experiment about 8 weeks after the last rat of the control group was affected with paralysis. The neural tumours induced in all groups were predominantly large tumours found in the cerebrum of the rats. ENU-treated rats showed a 100% incidence of nervous system tumours with a mean time of manifestation of neurological symptoms of 282 days, which was significantly shorter in comparison to that noted in the FBPI-treated group. The latter group showed an incidence of 58.3%, i.e. a significant reduction of 41% in the incidence of neural tumours, as well as a lower mean value for the number of tumours per rat. All these aspects indicated that FBPI is a potential neurooncopreventive agent. A neural tumour incidence of 100% in the rats treated with heat-inactivated FBPI confirmed that the tumour suppressive activity of FBPI is related to its protease inhibitory activity. PMID- 9751270 TI - Analysis of DNA methylation of the 5' region of the deoxycytidine kinase gene in CCRF-CEM-sensitive and cladribine (CdA)- and 2-chloro-2'-arabino-fluoro-2' deoxyadenosine (CAFdA)-resistant cells. AB - DNA methylation of the CpG-rich 5' region of the deoxycytidine kinase (dCK) gene is potentially involved in the suppression of the gene and the resistance of tumour cells to arabinosylcytosine (ara-C). 2-Chlorodeoxyadenosine (cladribine, CdA) and 2-chloro-2'-arabino-fluoro-2'-deoxyadenosine (CAFdA) are purine nucleoside analogues which are also phosphorylated by dCK. We observed a reduction in dCK activity in a number of CCRF-CEM-derived cell lines that are resistant to these drugs and hypothesized that this reduction is due to DNA methylation of the 5' region of the dCK gene. The DNA methylation state was analyzed at the DNA sequence level after bisulfite modification of genomic DNA. The investigated region included 0.3 kb of DNA upstream to the start site of transcription, exon 1 and part of intron 1. Sensitive cells (CCRF-CEM/0) and three resistant cell lines (CCRF-CEM/CdA4000, CCRF-CEM/CAFdA100 and CCRF CEM/CAFdA4000) were investigated. The region that was analyzed contained no methylated cytosine residues in the parental cell line CCRF-CEM/0 or in the resistant cell lines. Therefore, it is highly unlikely that DNA methylation plays a role in the suppression of dCK gene expression in these cell lines. PMID- 9751272 TI - Effect of vinblastine on cell membrane fluidity in vinblastine-sensitive and resistant HeLa cells. AB - The electron paramagnetic resonance method (EPR) was used to study the effects of vinblastine (VLB) on cell membrane fluidity in wild-type HeLa cells (HeLa K) and its subclone, which is resistant to several drugs (HeLa CA). HeLa CA cells, obtained by treatment of HeLa K with CDDP, were found to be more resistant to VLB than to CDDP. The experimentally observed EPR spectra were correlated with the calculated spectra obtained by computer simulation. The results indicate that the cell membrane of HeLa K and HeLa CA cells is heterogeneous and can be described with at least three types of coexisting domains with different fluidity characteristics. The two more fluid domains of HeLa CA cells were found to be more fluid than in HeLa K cells. The fluidity of the less fluid domain remained unchanged but its portion in the membrane was increased. VLB treatment did not affect the membrane fluidity of HeLa CA cells significantly. On the other hand, 1 h of treatment of HeLa K cells with 1 ng/ml VLB did not change the fluidity characteristics of membrane domains but decreased the portion of the less fluid domains. This was also reflected in an average increase of the entire membrane fluidity. The observed changes were detected at VLB concentrations which were far below the cytotoxic level. PMID- 9751271 TI - Increased sensitivity to vincristine of MDR cells by the leukotriene D4 receptor antagonist, ONO-1078. AB - The leukotriene D4 (LTD4) receptor antagonist, 4-oxo-8-[p-(4 phenylbutyloxy)benzoylamino]-2-(tetrazol-5-yl) -4H-1-benzopyran hemihydrate (ONO 1078) is used for the treatment of allergic asthma and other immediate hypersensitivity diseases. We examined the effect of ONO-1078 on the sensitivity to vincristine (VCR) of MRP overexpressing multidrug-resistant CV60 and its parental drug-sensitive KB-3-1 cell lines. The sensitivity to VCR of KB-3-1 and CV60 cells was increased 13- and 15-fold, respectively, by ONO-1078 at the maximum non-toxic concentration (100 microM). The VCR sensitivity of multidrug resistant KB-C2 cells that overexpressed P-gp was increased 2.6-fold by ONO-1078. The accumulation of VCR in KB-3-1, CV60 and KB-C2 cells was significantly increased by ONO-1078. The efflux of VCR from KB-3-1 cells was not inhibited, but that from CV60 cells was enhanced compared with that from KB-3-1 cells and was partially inhibited by ONO-1078. ONO-1078 competitively inhibited the ATP dependent [3H]LTC4 uptake in membrane vesicles isolated from CV60 cells. These findings suggest that ONO-1078 inhibits the transporting activity of MRP and that ONO-1078 increases the sensitivity to VCR of KB-3-1 cells by increasing the VCR uptake in the cells. PMID- 9751273 TI - Linoleic acid and tumor necrosis factor-alpha increase manganese superoxide dismutase activity in intestinal cells. AB - Manganese superoxide dismutase (MnSOD) protects mitochondria from oxidative damage. Alterations in the regulation of MnSOD plays an important role in the development of many types of cancer. Activity of this enzyme is induced by inflammatory cytokines and other conditions that increase oxygen radical production. High levels of dietary lipid have been shown to decrease MnSOD activity. This study was designed to define the effect of various type of fatty acids on MnSOD activity and MnSOD induction. IEC-6 cells were treated with 40 micromol/l of either linoleic acid (LA), eicosapentaenoic acid (EPA), or oleic acid (OA) in the presence or absence of 10 ng/ml tumor necrosis factor-alpha (TNF alpha). Fatty acid supplementation increased MnSOD activity. MnSOD activity was greater in the LA group than in the EPA or OA groups. TNF-alpha induced MnSOD activity equally in all fatty acid-supplemented groups. High levels of MnSOD activity may be an indicator of chronic inflammation resulting from fatty acid, particularly LA, supplementation. PMID- 9751274 TI - Cytotoxic and antitumour principles from Ixora coccinea flowers. AB - The antitumour activity of Ixora coccinea L. (Rubiaceae) flowers was studied in comparison to intraperitoneally transplanted Dalton's lymphoma (ascitic and solid tumours) and Ehrlich ascites carcinoma (EAC) tumours in mice. Intraperitoneal administration of 200 mg/kg of the active fraction (AF) of the I. coccinea flower increased the life-span of DLA and EAC ascitic tumour-bearing mice by 113 and 68%, respectively. The AF showed less activity against solid tumours (DLA) as compared to ascitic tumours. The same active fraction showed 50% cytotoxicity to DLA, EAC and Sarcoma-180 (S-180) cells in vitro at concentrations of 18, 60 and 25 microg/ml, respectively. It was not toxic to normal lymphocytes, whereas it was toxic to transformed lymphocytes from leukaemic patients, acute lymphoblastic leukaemia (ALL) and chronic myelogenous leukaemia (CML) and K-562 suspension cell cultures. The AF inhibited tritiated thymidine incorporation in cellular DNA. PMID- 9751275 TI - Familial pleural malignant mesothelioma: clustering in three sisters and one cousin. AB - Malignant mesothelioma is associated with asbestos, yet its occurrence in genetically-related individuals suggests a role of host predisposition. We have identified a cluster of pleural malignant mesothelioma in three sisters and one cousin (male). The women had worked in the same confectionery shop as pastry cooks and/or pastry shop assistants; the use of an asbestos-insulated oven was the putative source of exposure. The man had occupational exposure as a heating system installation worker. The family history reported malignant cancers in first-degree (larynx, brother; pleura and lung, mother), second-degree (lung, aunt and uncle) and third-degree (lung, cousin) relatives. The study reveals the potential existence of the mesothelioma risk among pastry cooks and highlights the fact that inherited factors may be involved in the development of this tumour. PMID- 9751276 TI - Estrogenic and antiproliferative activities on MCF-7 human breast cancer cells by flavonoids. AB - The interaction between the estrogen receptor and a variety of flavonoids was studied in the presence or absence of estradiol using a stably-transfected human breast cancer cell line (MVLN). On the other hand, flavonoids were evaluated for their effects on proliferation in estrogen-dependent (MCF-7) and independent (MDA MB231) human breast cancer cells. We established a relationship structure activity and determined regions and/or substituents essential for estrogenic or antiestrogenic activities. In contrast, we did not find the same relationship for cell proliferation. Among all flavonoids used, only 7-methoxyflavanone and 7,8 dihydroxyflavone at high concentrations (50 microM) possess antiestrogenic and antiproliferative activities. These results suggest that two hydroxyls (in positions 7 and 8) or 7-methoxy substituents are essential for the antiestrogenic activity of flavonoids. However, it seems that flavonoids at high concentrations exert their antiproliferative activity through other estrogen receptor independent mechanisms. PMID- 9751278 TI - Electrophysiological correlates of sleep delta waves. AB - Recent studies have disclosed several oscillations occurring during resting sleep within the frequency range of the classical delta band (0.5-4 Hz). There are at least 3 oscillations with distinct mechanisms and sites of origin: a slow (<1 Hz) cortically-generated oscillation, a clock-like thalamic oscillation (1-4 Hz), and a cortical oscillation (1-4 Hz). The present paper reviews data on these oscillations and the possible mechanisms which coalesce them into the polymorphic waves of slow wave sleep. Data stem from intracellular (over 500 single cell and 50 double impalements) and field potentials recorded from the cortex and thalamus of cats (120 animals) under ketamine and xylazine anesthesia. Other experiments were based on whole night EEG recordings from humans (5 subjects). The frequency of the slow oscillation both in anesthetized animals and in naturally sleeping humans ranged between 0.1 and 1 Hz (89% of the cases being between 0.5 and 0.9 Hz). The slow (<1 Hz) oscillation is reflected in the EEG as rhythmic sequences of surface-negative waves (associated with hyperpolarizations of deeply-lying neurons) and surface-positive K-complexes (representing excitation in large pools of cortical neurons). Through its long-range synchronization, the slow oscillation has the ability to trigger and to group thalamically-generated spindles and two delta (1-4 Hz) oscillations. Finally, it is argued that the analysis of the electroencephalogram should transcend the spectral analyses, by taking into account the shape of the waves and, when possible, the basic mechanisms that generate those waves. PMID- 9751277 TI - The effect of soy isoflavones on the development of intestinal neoplasia in ApcMin mouse. AB - Data from epidemiological studies suggest that isoflavones in soy may have a protective effect on the development of colon cancer in humans. Therefore, we have investigated whether soy isoflavones will inhibit intestinal tumour development in Apc(Min) mice. The mice were fed a Western-type high risk diet (high fat, low fibre and calcium) containing two different isolates of soy protein as a protein source. For the control and test groups this resulted in the administration of about 16 and 475 mg of total isoflavones per kg diet, respectively. As a positive control. a third group of mice was administered a low isoflavone diet supplemented with 300 ppm sulindac. No significant differences in the incidence, multiplicity, size and distribution of intestinal tumours were observed between Min mice fed low and high isoflavone-containing diets. However, a clear reduction in the number of small intestinal tumours was observed for the sulindac diet. Thus, in contrast to epidemiological studies, our results demonstrate that high amounts of soy isoflavones present in a Western-type high risk diet do not protect against intestinal tumour development in a relevant animal model such as the Min mice. PMID- 9751279 TI - Automatic auditory word perception as measured by 40 Hz EEG responses. AB - OBJECTIVES: Here we report the existence of automatic speech perception in man, revealed by 40 Hz EEG responses. METHODS: We presented to Finnish subjects the Finnish word /tu:li/(wind) as the standard stimulus and another Finnish word /tuli/(fire) as the deviant stimulus using a passive auditory oddball task. The experiment was also conducted with pseudowords as stimuli. RESULTS: We observed a global significant increase in 40 Hz EEG power at 600 ms after stimulus onset for words, but not for pseudowords. CONCLUSIONS: Our results indicate that the memory representation of the standard verbal stimuli, even if unattended, might not merely be based on the physical features of the stimuli: if a semantic representation exists, then the brain processes it pre-attentively. PMID- 9751280 TI - Spike topography and functional magnetic resonance imaging (fMRI) in benign rolandic epilepsy with spikes evoked by tapping stimulation. AB - We performed a spike topography study and a functional magnetic resonance imaging (fMRI) in a female patient with benign rolandic epilepsy presenting single high amplitude evoked spikes in response to somatosensory peripheral stimulation. The stimulus was delivered to the first finger of the right hand using a tendon hammer, which evoked a single spike followed by a slow wave, showing the maximal amplitude over the left central regions. fMRI showed that the contralateral sensory cortices (S1 and S2) and the motor cortex (M I) were activated during tapping stimulation. In 3 normal subjects, tapping stimulation produced no fMRI activation. This fMRI study documents a highly focal activation of sensorimotor areas related to subclinical evoked spikes in benign rolandic epilepsy. PMID- 9751281 TI - Improving source reconstructions by combining bioelectric and biomagnetic data. AB - OBJECTIVES: A framework for combining bioelectric and biomagnetic data is presented. The data are transformed to signal-to-noise ratios and reconstruction algorithms utilizing a new regularization approach are introduced. METHODS: Extensive simulations are carried out for 19 different EEG and MEG montages with radial and tangential test dipoles at different eccentricities and noise levels. The methods are verified by real SEP/SEF measurements. A common realistic volume conductor is used and the less well known in vivo conductivities are matched by calibration to the magnetic data. Single equivalent dipole fits as well as spatio temporal source models are presented for single and combined modality evaluations and overlaid to anatomic MR images. RESULTS: Normalized sensitivity and dipole resolution profiles of the different EEG/MEG acquisition systems are derived from the simulated data. The methods and simulations are verified by simultaneously measured somatosensory data. CONCLUSIONS: Superior spatial resolution of the combined data studies is revealed, which is due to the complementary nature of both modalities and the increased number of sensors. A better understanding of the underlying neuronal processes can be achieved, since an improved differentiation between quasi-tangential and quasi-radial sources is possible. PMID- 9751282 TI - Event-related desynchronization and synchronization during an auditory lexical matching task. AB - OBJECTIVES: Event-related desynchronization (ERD) and synchronization (ERS) of the 8-10 and 10-12 Hz frequency bands of the background EEG were studied in 10 subjects performing an auditory lexical matching task. METHODS: The stimuli were words and pseudowords presented sequentially in pairs. The subject was prompted to answer whether the two stimuli shared the same lexical status (words or pseudowords). RESULTS: Regardless of lexicality, the presentation of the first stimulus elicited a significant late frontal ERD in both alpha frequency bands. When preceded by a pseudoword, the presentation of the second stimulus elicited a significant ERS at 200-400 ms and a significant, long-lasting and topographically widespread ERD at 600-2200 ms in both frequency bands. When preceded by a word, the second stimulus did not elicit ERS in the initial time window, but a late ERD which was similar to the one observed in the previous condition. The complexity of ERD/ERS changes in the present task was revealed by significant interactions that time had with frequency band, stimulus type, stimulus order and lexicality of the preceding stimulus. CONCLUSIONS: The results suggest that ERD/ERS does not reflect primary auditory stimulus processing. Rather, the ERD/ERS observed in this experiment most probably reflected task difficulty and differences between lexical-semantic and phonological memory functions. PMID- 9751283 TI - Homeostatic process and sleep spindles in patients with sleep-maintenance insomnia: effect of partial (21 h) sleep deprivation. AB - OBJECTIVES: A low level of process 5 at bed time would be responsible for a reduced amount of slow-wave activity (SWA) leading to increased alpha activity and awakenings at the end of the night. METHODS: Following a base-line night (BLN) recording, 7 sleep-maintenance insomnia (SMI) subjects and 7 sex- and age matched controls were maintained on 21 h of sleep deprivation. Thereafter, a recovery night (RN) was performed from 2300 h until spontaneous awakening. SWA (power density of the EEG delta band between 0.75 and 4.5 Hz) was monitored by means of spectral analysis (FFT). Sleep spindles and the occupation ratio of Rechtschaffen and Kales EEG bands were observed by integrated digital filtering analysis. RESULTS: SWA was lower in SMI subjects than in controls during RN but was higher than in BLN indicating that the homeostatic process was operating, but weaker in SMI subjects. On the other hand in SMI subjects the sleep spindle index (SSI) did not decrease during slow-wave sleep and was significantly lower than in controls. Moreover during RN the SSI decreased significantly during the first sleep cycle in controls and not in SMI subjects. The existence of an inverse relationship between SWA and SSI was therefore not observed in insomniacs. Finally the mean duration of alpha frequency significantly increased in SMI subjects. CONCLUSIONS: It is hypothesised that in SMI subjects, an alteration of the homeostatic process is responsible for insufficient sleep pressure leading to an inability to maintain sleep for an extended period. PMID- 9751284 TI - Double magnetic stimulation of the motor cortex in amyotrophic lateral sclerosis. AB - OBJECTIVES: To study the motor cortex circuitry and the motor interhemispheric influences with double magnetic stimulation in patients affected by amystrophic lateral sclerosis. METHODS: We investigated the motor cortex in 21 amyotrophic lateral sclerosis patients (ALS, 10 with bulbar and 11 with spinal onset) with double magnetic stimulation (one shock in each hand area) with 2, 4, 6, 11 and 15 ms delay between shocks and paired magnetic stimulation (both shocks in the same area), with 4, 15, 25, 35, 55, 85, 100, 155, 200 and 255 ms delays, and compared the results with those obtained in normal subjects. RESULTS: Double magnetic stimulation showed reduced interhemispheric facilitatory influences (maximal at 4 ms delay between shocks) when the test shock was applied on the left hemisphere in all patients; whereas no significant differences were observed compared to control (P > 0.05) when it was applied on the right hemisphere in both forms. Inhibitory effects (maximal at 11 ms delay between shocks) were reduced in all patients for both hemispheres (P < 0.05). Paired magnetic stimulations showed decreased inhibitory influences at 100-155 ms delay between shocks. Compared to control, the difference was significant in bulbar (P < 0.05) and spinal onset, but not between onset forms (P > 0.05). Inhibitory effects recorded with a short delay between shocks (4 ms) were not significantly modified in both forms of onset (P > 0.05) as compared to control. There were no facilitatory influences at 15 and 35 ms delays between shocks. CONCLUSIONS: The results suggest that under these test conditions inhibition and facilitation were reduced in the motor cortex in ALS. As inhibitory effects were affected differently, two distinct cortical circuitries could be involved for short and long delays. As GABA neurons altered in ALS have been identified as a subpopulation reactive to parvalbumin, and since only inhibitory effects recorded with long delay between shocks were impaired in ALS, we suggest that this subpopulation of GABA neurons may be involved in the genesis of inhibitory effects recorded with a long delay between shocks. PMID- 9751286 TI - Detection of muscle artefact in the normal human awake EEG. AB - OBJECTIVES: A study was performed to investigate automatic detection of muscle artefact, using time domain and frequency domain methods. The evaluation focussed on epoch length and performance of detection. METHODS: EEG data were recorded in 21 normal adult subjects for 50 min during awake state. Investigated positions included central, temporal and parietal scalp electrodes. Expert annotation of muscle artefact was performed by accurate visual marking in a randomised test-set of the data, which allowed for intra-expert comparison. For time domain detection, the parameter set consisted of slope and maximum/minimum amplitude. Parameters in the frequency domain were absolute and relative 'high beta' power (>25 Hz) and spectral edge frequency. Distributions as calculated from a reference period in each subject were used to investigate the statistics of the parameter ranges. Detection thresholds were calculated from these distributions per subject, and performance was compared to constant (empirical) thresholds for the entire data set. RESULTS: Results indicate a 1 s epoch length as optimal for detection of muscle artefact. The analysis using a slope threshold or absolute 'high beta' power showed the best results in sensitivity (80%) and specificity (90%), matching the expert's performance. CONCLUSIONS: Constant threshold settings performed better than statistical thresholds per subject. PMID- 9751285 TI - Neurophysiological monitoring of pharmacological manipulation in acute organophosphate (OP) poisoning. The effects of pralidoxime, magnesium sulphate and pancuronium. AB - OBJECTIVES: The neuromuscular transmission failure in acute organophosphate (OP) poisoning occurs because of the irreversible inactivation of the enzyme acetylcholinesterase located in the neuromuscular junction, and is distinguished neuroelectrophysiologically by single electrical stimulus-induced repetitive responses and either a decremental or a decrement-increment response upon high rate repetitive nerve stimulation (RNS). Understandably, the administration of pharmacological agents with actions at different sites in the neuromuscular junction would alter the neuroelectrophysiological findings in acute OP poisoning. METHODS: The effect of several pharmacological agents including pralidoxime (10 patients), magnesium sulphate (4 patients) and pancuronium (7 patients) on the neuroelectrophysiological abnormalities was studied in 21 patients with acute OP poisoning. RESULTS: Pralidoxime administration produced neurophysiological amelioration in 11 out of 15 occasions. In those cases where it produced a beneficial effect, pralidoxime administration was continued and its neuroelectrophysiological effects were studied daily. The efficacy of pralidoxime administration was demonstrated by neuroelectrophysiological testing for a maximum of 6 days after poisoning. Three types of neuroelectrophysiological responses to pralidoxime were noted: (i) lack of neuroelectrophysiological improvement (two patients); (ii) initial improvement with subsequent lack of improvement (two patients); and (iii) initial improvement with subsequent normalisation of neuromuscular transmission (5 patients). Normalisation of the electrodiagnostic tests and the failure of pralidoxime to ameliorate the neuromuscular transmission abnormalities were neuroelectrophysiological indications for the discontinuation of pralidoxime treatment. The administration of magnesium sulphate (MgSO4.7H2O, 4 g intravenous) resulted in a decrease in the CMAP amplitude, loss of the repetitive response and conversion of the decrement increment response at high-rate RNS to an incremental response. Repetitive responses and the decremental response at high-rate RNS also disappeared after the administration of pancuronium (0.5 mg intravenous) to 6 patients. However, in one case where pancuronium administration was preceded by pralidoxime, there occurred a dramatic worsening of the neuromuscular transmission defect. CONCLUSIONS: While the administration of all 3 agents-- pralidoxime, magnesium sulphate and pancuronium-- resulted in the reversion of the neuroelectrophysiological defects, only pralidoxime is contended to be therapeutically useful. The therapeutic benefit due to its administration is limited by a short duration of action, and hence it is recommended that it should be administered for a longer period of time under neuroelectrophysiolgical guidance. PMID- 9751287 TI - A study of dipole localization accuracy for MEG and EEG using a human skull phantom. AB - OBJECTIVE: To investigate the accuracy of forward and inverse techniques for EEG and MEG dipole localization. DESIGN AND METHODS: A human skull phantom was constructed with brain, skull and scalp layers and realistic relative conductivities. Thirty two independent current dipoles were distributed within the 'brain' region and EEG and MEG data collected separately for each dipole. The true dipole locations and orientations and the morphology of the brain, skull and scalp layers were extracted from X-ray CT data. The location of each dipole was estimated from the EEG and MEG data using the R-MUSIC inverse method and forward models based on spherical and realistic head geometries. Additional computer simulations were performed to investigate the factors affecting localization accuracy. RESULTS: Localization errors using the relatively simpler locally fitted sphere approach are only slightly greater than those using a BEM approach. The average localization error over the 32 dipoles was 7-8 mm for EEG and 3 mm for MEG. CONCLUSION: The superior performance of MEG over EEG appears to be because the latter is more sensitive to errors in the forward model arising from simplifying assumptions concerning the conductivity of the skull, scalp and brain. PMID- 9751288 TI - Usefulness of latero-orbital electrodes in detecting interictal epileptiform activity--a study of 60 patients with complex partial seizures. AB - OBJECTIVES: The use of latero-orbital (Lo) electrodes is a routine practice in any EEG laboratory to evaluate eye motion, but there are no data about their usefulness in revealing interictal epileptiform abnormalities. METHODS: In 60 consecutive patients (27 men, 33 women, mean age 36.8 years, range 17-72) with complex partial seizures, we prospectively evaluated the utility of Lo electrodes in comparison with anterior temporal (AT) electrodes, for the detection of interictal epileptiform discharges (SW). RESULTS: No epileptiform abnormality was seen in 4/60 patients. Both AT and Lo electrodes were significantly superior to 10-20 electrodes for detection of both patients and foci. Indeed, the standard 10 20 system alone allowed the detection of only 39 independent epileptiform foci in 35/56 (63%) patients, while AT and Lo electrodes were necessary for detection of 23 epileptiform foci in the remaining 21/56 (37%) patients. Importantly, there was no statistically significant difference in detection between AT and Lo electrodes. CONCLUSIONS: Recordings from Lo electrodes are comparable to those from AT electrodes and are useful for localizing interictal temporal spiking activity. Lo electrodes may be substituted for basal electrodes in the day-to-day evaluation of patients with complex partial seizures. PMID- 9751289 TI - Long-latency response to transcranial magnetic stimulation in patients with hemifacial spasm. AB - OBJECTIVE: We studied the long-latency response of the orbicularis oris muscle elicited with transcranial magnetic stimulation in patients with hemifacial spasm (HFS) and evaluated the excitability of the facial nucleus. METHODS: We compared the thresholds on both sides in 8 normal volunteers and 7 patients with hemifacial spasm. The thresholds were determined as the lowest intensity required to produce motor evoked potentials with an amplitude of at least 50 microV in the orbicularis oris muscle. Average values were given as means +/- standard deviation. Wilcoxon's rank sum test was used for comparisons between the sides of normal subjects and of patients with HFS with respect to the threshold stimulus. RESULTS: There was no significant difference between the thresholds on the two sides of the normal subjects (mean 1.88+/-5.30%, P > 0.05). In patients with HFS, there was a significant difference between the thresholds on the spasm side and the normal side (mean 20.7+/-13.0%, P < 0.05) In one patient studied after MVD, the difference between both sides disappeared. CONCLUSION: The difference between the thresholds in patients with HFS and the normalization in threshold after MVD suggested that the mechanism of HFS was hyperexcitability of the facial nucleus. PMID- 9751290 TI - Reproducibility of electromyographic recordings of submaximal concentric and eccentric muscle contractions in humans. AB - OBJECTIVES: to determine the intratester and intertester reliability of measures of electromyographic activity (EMG) of submaximal concentric and eccentric contractions and to compare the reliability of normalized and non-normalized measures of EMG. There were 10 subjects, of 22-33 years old. METHODS: Subjects performed submaximal concentric and eccentric contractions at 60 degrees/s, and maximal isometric contractions (MIC) of their knee extensors. The target power of the submaximal contractions was 90%+/-10% of the subject's maximal concentric power. EMG was recorded via bipolar surface electrodes from 3 of the quadriceps femoris muscles. The rmsEMG for submaximal contractions that were within the target power range were determined. The rmsEMG for the submaximal contractions were normalized to the rmsEMG of the maximal isometric contractions. Intraclass correlation coefficients (ICC version 1.1) were calculated to determine intratester and intertester reliability. RESULTS: For non-normalized rmsEMG, ICC values for intratester reliability ranged from 0.62 to 0.91 for concentric and from 0.84 to 0.97 for eccentric contractions. ICC values for intertester reliability ranged from 0.66 to 0.96 for concentric and 0.78 to 0.90 for eccentric contractions. CONCLUSIONS: Non-normalized rmsEMG of submaximal concentric and eccentric isokinetic contractions were found to be reliable. Normalization did not lead to consistently improved reliability. PMID- 9751291 TI - Latency of changes in spinal motoneuron excitability evoked by transcranial magnetic brain stimulation in spinal cord injured individuals. AB - OBJECTIVES: To examine the basis for delay in the excitatory effects of transcranial magnetic stimulation (TMS) of motor cortex on motoneuron pools of muscles left partially-paralyzed by traumatic spinal cord injury (SCI). METHODS: The effect of subthreshold transcranial magnetic stimulation (TMS) on just suprathreshold H-reflex amplitude was examined in subjects (n = 10) with incomplete cervical SCI, and in able-bodied (AB) subjects (n = 20) for comparison. EMG activity was recorded from the soleus and the abductor hallucis muscles, and H-reflex was elicited by stimulation of the tibial nerve behind the knee. Comparison of the peak-to-peak amplitude of the TMS-conditioned H-reflex to that of the H-reflex alone (i.e. unconditioned H-reflex) was made for different conditioning-test intervals with multivariate analysis of variance and (when called for) t testing. RESULTS: The absolute latencies of motor responses to suprathreshold TMS delivered during a weak voluntary contraction of the soleus and abductor hallucis were significantly prolonged in the SCI group relative to AB subjects. For the TMS-conditioned H-reflex, the time-course effect of TMS on the H-reflex amplitude in different AB subjects included an early effect (typically facilitation, but occasionally inhibition) seen between -5 and 0 ms, followed by a later period (i.e. >5 ms) of H-reflex facilitation. In contrast, the earliest indication of a TMS effect on H-reflex excitability in SCI subjects was between 5 and 10 ms after TMS. This difference between SCI and AB subjects of approximately 10 ms was similar to the prolongation of TMS-evoked response latencies in the soleus and the abductor hallucis muscles of the SCI subjects. CONCLUSIONS: The results suggest that motor conduction slowing after traumatic SCI most likely occurs across the population of the descending tract axons mediating the TMS-evoked motor responses. PMID- 9751292 TI - Carpal tunnel syndrome in childhood: study of 6 cases. AB - Six children, 4 girls and two boys, aged 5-14 years, with carpal tunnel syndrome (CTS) are reported. Median nerve entrapment had different aetiologies in each case. One patient developed unilateral CTS symptoms after intensive basketball training. He improved upon terminating this sporting activity. In 3 patients bilateral CTS was associated with Schwartz-Jampel syndrome, trigger finger and mucopolysaccharidosis I (MPS IS = Scheie syndrome), respectively. The latter subject, a boy aged 11 years who had severe bilateral muscle thenar weakness and atrophy, made a good recovery after surgery. Two cases with bilateral CTS had autosomal dominant disease. One of them showed familial CTS with thickening of the transverse carpal ligament. The other child (5 years old) presented early bilateral CTS as first manifestation of hereditary neuropathy with liability to pressure palsies (HNPP). His relatives were asymptomatic, but they showed electrophysiological and nerve biopsy changes consistent with HNPP. Nerve conduction studies (NCS) are diagnostic in paediatric CTS. Moreover, NCS is an objective method to evaluate the evolution of the nerve lesions after surgery. NCS must be performed in nerves of the propositus other than the median, as well as in first degree symptomatic and asymptomatic relatives in order to identify possible familial neuropathies. PMID- 9751293 TI - Responses of masseter muscles to transcranial magnetic stimulation in patients with amyotrophic lateral sclerosis. AB - We recorded motor responses evoked by transcranial magnetic stimulation (TMS) in the masseter muscles of 30 patients with amyotrophic lateral sclerosis (ALS), 10 patients with cervical spondylotic myelopathy (CSM) and 22 age-matched normal controls. Responses to direct activation of the trigeminal motor root (R-MEPs) were normal both in ALS and CSM patients. Responses to activation of cortico bulbar descending fibers (C-MEPs) were absent or delayed in 19 ALS patients (63.3%). Abnormalities of masseter C-MEPs were more frequent than abnormalities of limb MEPs and could be observed both in patients with (77.8%) and without (41.7%) clinical bulbar signs. Masseter C-MEPs were normal in all CSM patients. Recording masseter responses to TMS can reveal the frequent impairment of cortico bulbar projections in ALS and can be useful in the differential diagnosis of spinal cord compression disorders mimicking ALS because of combination of upper and lower motor neuron signs. PMID- 9751294 TI - Magnetoneurographic 3D localization of conduction blocks in patients with unilateral S1 root compression. AB - OBJECTIVES: Tibial nerve somatosensory evoked magnetic fields (tSEFs) over the lower back reflect the propagation of compound action currents along fibers of plexus, nerve roots and cauda equina. One clinical perspective for this 'magnetoneurography' is the non-invasive 3D localization of focal slowing or blocks of conduction. Here, first tSEF mappings in 3 consecutive patients with acute unilateral S1 nerve root compression are reported. METHODS: Right and left tibial nerves were electrostimulated in alternation; tSEF responses were recorded using a multichannel SQUID-detector; additionally, spinal and cortical SEP, F wave and H-reflex studies were performed. RESULTS: In all patients an intraindividual side-to-side comparison of spinal tSEF mappings was obtained: using a dipolar source model compound action currents could be visualized propagating along plexus, nerve roots and cauda equina on the non-affected side whereas on the affected side normally-propagating dipolar field patterns could be recorded only distal to the spinal transforaminal root entrance; this reflects focal slowing or block of conduction in nerve root fibers as indicated by the SEP, F-wave and H-reflex study results. CONCLUSIONS: With a registration time of 15 min a 3D localization of proximal slowing or block of conduction was successfully performed in patients suffering from acute nerve root lesions. PMID- 9751295 TI - Pharmacological control of facilitatory I-wave interaction in the human motor cortex. A paired transcranial magnetic stimulation study. AB - A novel paired transcranial magnetic stimulation (TMS) paradigm with a suprathreshold first and a subthreshold second stimulus was used in healthy volunteers to investigate the acute effects of a single oral dose of various CNS active drugs on short-interval motor evoked potential (MEP) facilitation. MEPs were recorded from the relaxed abductor digiti muscle. Three peaks of MEP facilitation were consistently observed at interstimulus intervals of 1.1-1.5 ms, 2.3-2.7 ms, and 3.9-4.5 ms. The size of these MEP peaks was transiently suppressed by drugs which enhance gamma-aminobutyric acid (GABA) function in the neocortex (lorazepam, vigabatrin, phenobarbital, ethanol), while the GABA-B receptor agonist baclofen, anti-glutamate drugs (gabapentin, memantine), and sodium channel blockers (carbamazepine, lamotrigine) had no effect. The interstimulus intervals effective for the production of the MEP peaks remained unaffected by all drugs. The MEP peaks are thought to be due to a facilitatory interaction of I-(indirect) waves in the motor cortex. Therefore, the present results indicate that the production of I-waves is primarily controlled by GABA related neuronal circuits. The potential relevance of this non-invasive paired TMS protocol for the investigation of I-waves in patients with neurological disease will be discussed. PMID- 9751296 TI - Electrodiagnostic methods for neurogenic dysphagia. AB - OBJECTIVE: Swallowing mechanisms and neurogenic dysphagia have not been systematically studied by the EMG technique. It is desirable to evaluate neurogenic dysphagia for diagnostic and possibly for therapeutic purposes using electrophysiological methods. METHODS: The following methods were described: mechanical upward/downward movements of the larynx were detected using a piezoelectric sensor, while submental integrated EMG activity was recorded during dry and wet swallowing. The EMG activity of cricopharyngeal muscle of the upper oesophageal sphincter was also recorded in some normal subjects and patients. Piecemeal deglutition and the dysphagia limit were determined in all patients to detect dysphagia objectively. In this study 75 normal subjects and 177 neurological patients with various degrees of dysphagia were investigated. RESULTS: Voluntarily triggered oropharyngeal swallowing was commonly pathological in the majority of patients, with or without overt dysphagia. The dysphagia limit appeared to be an objective measure of the degree of dysphagia in more than 90% of patients. Pathophysiological mechanisms were different in at least three groups of patients with neurogenic dysphagia. In the group of patients with muscular disorders, laryngeal elevators were involved while the CP-sphincter was intact. The second group included patients with the clinical signs of corticobulbar fibre involvement such as amyotrophic lateral sclerosis and pseudobulbar palsy. In these patients, there was incoordination between paretic laryngeal elevators and hyperreflexic CP-sphincter. In the third group (patients with Parkinson's disease), the swallowing reflex was delayed and prolonged. CONCLUSIONS: EMG methods described in the present study are very useful for the diagnosis of neurogenic dysphagia, objectively and quickly. They are important to understand the physiological mechanisms for deglutition and its disorders. PMID- 9751297 TI - Time dependent selective recurrent discharge of motor units in F-response. AB - The interaction among the recurrent discharge of motor units is studied in surface recorded composite F-response. In the first experiment, 200 serially elicited polyphasic F-responses from foot muscles of patients with different lower motor neurone (LMN) disorders were rearranged to understand the behaviour of individual negative and positive peaks. These peaks were considered on the basis of simultaneous recording with a single fibre EMG needle, to be the partial expression of either a single motor unit potential (MUP) or more than one MUP generated simultaneously. In another experiment, two serially averaged F responses (50 each) were superimposed over each other to look for their similarities at 15 min intervals 6 times. Result analysis indicated interaction of MUPs by in-phase summation and out-of-phase subtraction. Less affected MUPs recurred as negative or positive peaks in morphologically different F-responses. Certain peaks were observed more frequently than the others. Two serially averaged F-responses were nearly identical on superimposition over each other but their morphologies differed at different time intervals. The study suggests more frequent generation of F-response in a specific group of alpha motoneurones at one point of time, which is replaced by another group at another point of time. The characteristic variation in F parameters is, however, mainly due to the interaction of these frequently generated F-response MUPs with other sporadically generated ones. This interaction can be appreciated on visual screening of F responses in patients with lower motor neurone disorders, perhaps because of a reduction in the number of participating MUPs. PMID- 9751298 TI - Anticipatory postural adjustments in conditions of postural instability. AB - OBJECTIVES: The purpose of this study was to investigate anticipatory postural adjustments (APAs) in standing subjects who performed a standard motor action triggering a standard postural perturbation (releasing a 2.2 kg load from extended arms) in conditions of different stability requirements. METHODS: The degree of stability was varied either by balancing on special boards with long and narrow support beams or by instructions to the subjects. In the first series of experiments 13 subjects stood on the board facing either perpendicular to the beam (instability in a sagittal plane) or along the beam (instability in frontal plane); different widths of the beam were used to vary the degree of instability. During the second series of experiments (6 subjects) inclined and one-legged postures were used to induce instability in sagittal and frontal planes respectively. EMG activity of rectus abdominis, erector spinae, rectus femoris, biceps femoris, tibialis anterior, and soleus muscles were recorded. Statistical methods included repeated measures analysis of variance (ANOVA) with direction of instability and level of instability being major factors, descriptive statistics, and post hoc Student's t tests. RESULTS: The integral measure of changes in the background electromyographic activity of postural muscles during APAs depended on two factors related to the postural task: (1) standing on a platform with a narrow support area led to an attenuation of the APAs; and (2) these effects were stronger when instability was in a sagittal rather than in the frontal plane. The anticipatory component in the displacement of the center of pressure did not show a clear attenuation that would depend on the direction of instability. CONCLUSIONS: We suggest a hypothesis that, in conditions of high stability demands, the central nervous system may suppress APAs as a protection against their possible destabilizing effects. These effects are more pronounced when the direction of an expected perturbation is in the plane of instability. PMID- 9751299 TI - Coupling between single muscle spindle afferent and EMG in human wrist extensor muscles: physiological evidence of skeletofusimotor (beta) innervation. AB - OBJECTIVES: This study was conducted to find physiological evidence of skeletofusimotor innervation in man. METHODS: Discharges of 38 single muscle spindle afferents from m. extensor carpi radialis were recorded from 9 subjects during steady isometric contractions of wrist extension. Correlation between these afferents and rectified surface EMG was investigated by estimating cumulant density function. RESULTS: In the cumulant density estimate between spindle afferent and EMG, a positive EMG peak was obtained prior to afferent firing between -30 and -10 ms in 15 afferents (39%). CONCLUSIONS: The present finding indicates coupling between spindle afferents and extrafusal activity and suggests the widespread skeletofusimotor innervation in human muscle spindle. PMID- 9751300 TI - Cortical motor neuron excitability during cutaneous silent period. AB - OBJECTIVE: To investigate cortical motor neuron excitability during cutaneous silent period (CSP), motor evoked potentials (MEPs) from abductor pollicis brevis following transcranial magnetic stimulation (TCM) were recorded with and without a conditioning of ipsilateral painful digital nerve electric stimulation. METHODS: MEPs following TCM were recorded with and without a conditioning stimulation at an interstimulus interval (ISI) from 0 ms to 100ms in 6 controls and four patients who had reduced pain sensation in unilateral upper limbs associated with cervical syringomyelia. In addition MEPs and evoked spinal cord potentials (ESCPs) from cervical epidural space following TCM with and without a conditioning stimulation were recorded in four patients with thoracic myelopathy. RESULTS: MEP amplitude was clearly attenuated by a conditioning stimulation at an ISI from 40 ms to 80 ms in controls (statistically significant at 60 ms). In patients with cervical syringomyelia, MEP amplitude was attenuated by a conditioning stimulation in asymptomatic hands similarly in controls but that was unchanged by a conditioning stimulation in the symptomatic hand with reduced pain sensation. In patients with thoracic myelopathy MEP amplitude was attenuated by conditioning stimulation similarly in controls, but ESCP amplitude was unchanged. CONCLUSIONS: We demonstrated that noxious cutaneous nerve stimulation suppressed spinal motor neurons but cortical motor neuron excitability was unchanged during CSP. In clinical practice, measurement of MEP suppression after noxious cutaneous nerve stimulation may provide useful information in patients with damaged pain related nerve fibers. PMID- 9751301 TI - Peculiarity of soleus motor potentials evoked by transcranial magnetic stimulation and electrical stimulation of tibial nerve. AB - OBJECTIVE: To reveal and discuss the peculiarities of soleus muscle in comparison with electrophysiological features of other leg muscles. METHODS: Vastus lateralis (L3), tibialis anterior (L4), extensor digitorum brevis (L5) and soleus (S1) muscles were tested at rest. Transcranial magnetic stimulation (TMS) combined which electrical stimulation of relevant peripheral nerves were applied. Cortically evoked motor potentials (C-MEP), peripheral compound muscle action potential (CMAP) and F-wave were recorded. Estimating F-wave conduction time allowed to calculated the central conduction time (CCT-F) within the cortex spinal motoneurones segment for each muscle. RESULTS: One could expect that the lower spinal metameric representation of the muscle the longer a corresponding CCT-F. However, a study of 30 healthy subject (60 right and left muscles) reveals a relatively short CCT-F for the soleus muscle. Moreover, the mean amplitude of soleus C-MEP is the lowest, CMAP and F-wave amplitude are the highest and standardised distal motor latency is the longest compared to the analogous parameters for the other muscles. CONCLUSIONS: The reason for these special features can be attributed probably to a different structure and innervation of the soleus (mainly red, slow, tonic muscle) in contrast to the tibialis anterior and extensor digitorum brevis (mainly white, fast, phasic muscle). PMID- 9751302 TI - Effect of electrical stimulation of the thalamic Vim nucleus on hand tremor during stereotactic thalamotomy. AB - OBJECTIVE: The aim of this study was to analyze the correlation between neuronal responses in the thalamic ventralis intermedius (Vim) nucleus to peripheral, natural stimulation and the modulation of tremor by electrical stimulation during stereotactic thalamotomy. DESIGN AND METHODS: The authors studied 36 patients with hand tremor using a microelectrode. The responses of tremor to electrical stimulation were analysed electromyographically. Sixty stimulation sites were divided into three groups. RESULTS: Group A (20 sites) where responses to stretching of the contralateral forearm muscles were recorded. Group B (26 sites) where responses to stretching of the other muscles of the upper extremity were recorded. Electrical stimulation at sites in groups A and B temporarily suppressed the contralateral tremor, but the minimum current intensity to suppress tremor at sites in group A was less than that in group B. Electrical stimulation in group C (14 sites), where kinesthetic responses of contralateral lower extremity were recorded, resulted in increased amplitude of hand tremor. Selective coagulation including the area of tremor suppression resulted in abolition of the tremor in all patients. CONCLUSIONS: These results suggest that the most effective site for thalamotomy may also be suitable for chronic stimulation in the Vim nucleus. PMID- 9751322 TI - Commentary on metalloproteinases and bladder carcinoma. PMID- 9751323 TI - Management of hormone refractory prostate cancer: current standards and future prospects. AB - PURPOSE: Recent advances in the biology and treatment of hormone refractory prostate cancer are reviewed. MATERIALS AND METHODS: A MEDLINE literature search of secondary hormonal therapy and chemotherapy for hormone refractory prostate cancer was performed. Recent advances in the biology of hormone refractory prostate cancer, changes in the measurement of response to therapy, and testing of new drugs and combinations of drugs were reviewed. RESULTS: Historically the treatment of hormone refractory prostate cancer has been disappointing. Useful parameters to monitor clinical response have been lacking but perhaps more importantly a scarcity of apparently active drugs has contributed to these results. Recently several developments have improved the outlook for treatment of hormone refractory prostate cancer. Recognition of antiandrogen withdrawal responses has had important ramifications for clinical trial interpretation and patient care. Secondary hormonal therapies, such as alternative antiandrogens and anti-adrenal agents, are well tolerated and can provide significant clinical benefits. Combining prostate specific antigen values with quality of life and measurable disease responses has made clinical trial end points more objective and more clinically relevant for the patient. Furthermore, a better understanding of the biology of hormone refractory prostate cancer, refinements in measuring response to treatment and availability of agents with proved palliative capabilities and/or generating greater than 50% response have all lead to improvements in treatment management. In 2 randomized studies mitoxantrone in combination with steroids has demonstrated significant palliative benefit compared with steroids alone. In phase II studies more than half of patients respond to estramustine combinations with vinblastine, etoposide or paclitaxel. Other novel combinations and new drugs currently are being tested. CONCLUSIONS: Recent advances suggest that available therapies for hormone refractory prostate cancer can have a meaningful impact on the disease. Improving treatment of hormone refractory prostate cancer remains an area of active investigation. PMID- 9751324 TI - Progress in management of advanced prostate cancer--don't throw the baby out with the bath water. PMID- 9751325 TI - Renal autotransplantation for the loin pain-hematuria syndrome: long-term followup of 26 cases. AB - PURPOSE: We review the long-term results of renal autotransplantation as a form of nephron sparing renal denervation for patients with the loin pain-hematuria syndrome. MATERIALS AND METHODS: From 1985 to 1997, after exclusion of other urological, nephrological and psychiatric causes for severe intractable flank pain and recurrent hematuria, 22 patients with severe debility and heavy narcotic dependency underwent 26 renal autotransplantations for pain control. Postoperative pain relief, narcotic use, level of function in daily activities and status of the autograft were assessed. RESULTS: Median and mean followup was 78.5 and 84.7 months (range 30 to 138), respectively. There were 2 technical failures. Pain recurred within 2 years after 6 procedures, of which 3 resulted in transplant nephrectomy and 3 were managed with a reduced analgesic requirement. Of the 16 patients with minimum 5 years of followup 12 (75%) were pain-free after surgery with 3 additional patients pain-free after transplant nephrectomy. Overall, 18 of the 26 autotransplant procedures (69.2%) resulted in pain relief, in some cases beyond 10 years, with patients returning to normal daily activities. CONCLUSIONS: Renal autotransplantation results in durable narcotic free favorable results in the majority of meticulously screened loin pain hematuria syndrome patients. Although some failures, which usually occur within 2 years after surgery, can be expected, autotransplantation is justified as a nephron sparing denervation therapy for select loin pain-hematuria syndrome patients. PMID- 9751326 TI - Image based renal stone tracking to improve efficacy in extracorporeal lithotripsy. AB - PURPOSE: We describe a method to reduce the number of shocks necessary to fragment renal stones during extracorporeal shock wave lithotripsy by automatically taking into account stone movements. MATERIALS AND METHODS: Echotrack computer software was developed and implemented on a lithotriptor. One software module uses image processing to detect instantaneous stone location based on ultrasound images generated by the lithotriptor. A second module uses the detected location to control the shock wave generator position, and automatically adjusts it to improve coincidence between the focal volume and stone. The reliability of the tracking algorithm was clinically tested in 65 patients with renal stones. These in vivo tests were qualitative and the goal was to assess software ability to track stones during actual treatments. A quantitative evaluation of the reduction in shocks necessary for fragmentation was performed in vitro. Artificial stones were moved according to computer generated trajectories. Each trajectory was applied once with and once without automatic adjustment of the generator position. RESULTS: The in vivo tests demonstrated software ability to track stones as far as they were visible in the images. During in vitro tests automatic adjustments of the generator position reduced the number of shocks necessary to fragment stones completely by a factor of 1.64. CONCLUSIONS: Image based renal stone tracking software that automatically adjusts the shock wave generator position according to the displacement of renal stones is useful during extracorporeal shock wave lithotripsy. Treatment time was significantly shorter with this software. PMID- 9751327 TI - A randomized controlled trial to assess the incidence of new onset hypertension in patients after shock wave lithotripsy for asymptomatic renal calculi. AB - PURPOSE: To answer the question of whether extracorporeal shock wave lithotripsy (ESWL*) induces hypertension, a prospective, randomized controlled trial of normotensive patients with asymptomatic renal calculi was designed. MATERIALS AND METHODS: Patients were randomized to receive immediate ESWL versus observation, reserving ESWL for the onset of symptoms. The rates of new onset hypertension were evaluated for both groups. RESULTS: There was no observed difference in the incidence of hypertension between the treatment and observation groups. CONCLUSIONS: The risk of hypertension in patients undergoing ESWL therapy is similar to that of a control cohort of initially observed asymptomatic patients. PMID- 9751328 TI - Kidney transplantation: the use of living donors with renal artery lesions. AB - PURPOSE: A shortage of organs for transplantation has forced surgeons to optimize the use of marginal organs, such as kidneys with arterial disease. We present a retrospective study of the outcome of donors with renal artery disease and recipients of kidneys from living related and unrelated donors. MATERIALS AND METHODS: Kidneys with vascular abnormalities from healthy living donors were grafted into 11 recipients. These kidney transplants comprised 1.8% of those performed at our institution. The vascular abnormalities were aneurysms in 3 cases, atherosclerotic lesions in 4 and fibromuscular dysplasia in 4. After nephrectomy all abnormalities were corrected under hypothermic conditions during bench surgery except in 3 cases of ostial atherosclerotic plaque, which was left in the donors. The renal artery was anastomosed to the external iliac artery in 5 cases and to the internal iliac artery in 6. The ureter was reimplanted using an extravesical technique. RESULTS: All patients had immediate diuresis and no delayed post-transplant graft dysfunction was observed. One patient died of an unrelated cause and 3 had post-transplant graft function loss due to acute vasculopathy in 1, post-diarrhea with acute arterial thrombosis in 1 and recurrence of the hemolytic-uremic syndrome in 1. All remaining patients are well with median serum creatinine of 1.4 mg./dl. (normal 0.4 to 1.4). All donors are well and normotensive with normal renal function. CONCLUSIONS: The use of kidneys with arterial disease from living donors with unilateral disease is safe. Complete informed consent regarding the risks and benefits by donor and recipient is mandatory. PMID- 9751329 TI - Prevalence, morphology and biology of renal cell carcinoma in von Hippel-Lindau disease compared to sporadic renal cell carcinoma. AB - PURPOSE: Renal cell carcinoma occurs as a sporadic tumor but may be part of the autosomal dominant von Hippel-Lindau disease, characterized by retinal and central nervous system hemangioblastoma, pheochromocytoma, pancreatic cysts and renal cell carcinoma. We determine the prevalence of von Hippel-Lindau disease in a series of unselected renal cell carcinoma cases by molecular genetic analysis, and compare sporadic to von Hippel-Lindau renal cell carcinoma with respect to morphology and biology. MATERIALS AND METHODS: We established registers comprising 63 subjects with von Hippel-Lindau renal cell carcinoma, belonging to 30 distinct families (register A), and 460 unselected patients operated on for renal cell carcinoma in an 11-year period (register B). Molecular genetic analysis of the von Hippel-Lindau gene was performed for living patients of register A, representing 80% of von Hippel-Lindau families, and register B, 62% living patients, to identify von Hippel-Lindau germline mutations. In addition, register B was evaluated by a questionnaire (95% response) for familial occurrence of von Hippel-Lindau disease. RESULTS: The prevalence of von Hippel Lindau renal cell carcinoma was 1.6% in 189 consenting unselected renal cell carcinoma patients. Risk factors for occult germline von Hippel-Lindau gene mutations in register B included familial renal cell carcinoma in 3 of 3 patients (100%), multifocal or bilateral renal cell carcinoma in 1 of 10 (10%) and age younger than 50 years at diagnosis in 1 of 33 (3%). Compared to sporadic von Hippel-Lindau renal cell carcinoma was characterized by an occurrence 25 years earlier, association with renal cysts, multifocal and bilateral tumors, cystic organization and low grade histology, and a better 10-year survival (p < 0.001 each). In von Hippel-Lindau disease metastases occurred only in tumors larger than 7 cm. CONCLUSIONS: von Hippel-Lindau differs from sporadic renal cell carcinoma in morphology and biology. Our data provide arguments for planning surgery for von Hippel-Lindau renal cell carcinoma and should stimulate future investigations. PMID- 9751330 TI - Multiple primary malignancies in renal cell carcinoma. AB - PURPOSE: We determine the incidence and nature of multiple primary malignancies in patients with renal cell carcinoma, and whether these patients have an increased risk of a second primary malignancy. MATERIALS AND METHODS: Between July 1989 and January 1997, 551 patients underwent an operation for renal cell carcinoma. The incidence of other primary malignancies was determined and classified as antecedent, synchronous or subsequent. The observed number of subsequent malignancies after diagnosis of renal cell carcinoma was compared to the expected number based on age, race and sex specific 1990 to 1994 incidence rates from the United States Surveillance, Epidemiology and End Results data using the Poisson test. RESULTS: The number of primary malignancies, including cutaneous malignancies, was at least 1 in 148 patients (26.9%), at least 2 in 34 (6.2%), at least 3 in 6 (1.1%) and 4 in 1 (0.2%). Other malignancies were antecedent in 85 cases (45.0%), synchronous in 74 (39.4%) and subsequent in 30 (16.0%). The most common other primary malignancies were breast, prostate, colorectal and bladder cancer, and non-Hodgkin's lymphoma. Only men with renal cell carcinoma had an increased risk of bladder cancer (standardized incidence ratio 4.3, p = 0.0067). CONCLUSIONS: Breast, prostate, colorectal and bladder cancer as well as non-Hodgkin's lymphoma were the most common other primary malignancies. Men with renal cell carcinoma have an increased risk of subsequent bladder cancer. PMID- 9751331 TI - Optimal method of urgent decompression of the collecting system for obstruction and infection due to ureteral calculi. AB - PURPOSE: We compare the efficacy of percutaneous nephrostomy with retrograde ureteral catheterization for renal drainage in cases of obstruction and infection associated with ureteral calculi. MATERIALS AND METHODS: We randomized 42 consecutive patients presenting with obstructing ureteral calculi and clinical signs of infection (temperature greater than 38 C and/or white blood count greater than 17,000/mm.3) to drainage with percutaneous nephrostomy or retrograde ureteral catheterization. Preoperative patient and stone characteristics, procedural parameters, clinical outcomes and costs were assessed for each group. RESULTS: Urine cultures obtained at drainage were positive in 62.9% of percutaneous nephrostomy and 19.1% of retrograde ureteral catheterization patients. There was no significant difference in the time to treatment between the 2 groups. Procedural and fluoroscopy times were significantly shorter in the retrograde ureteral catheterization (32.7 and 5.1 minutes, respectively) compared with the percutaneous nephrostomy (49.2 and 7.7 minutes, respectively) group. One treatment failure occurred in the percutaneous nephrostomy group, which was successfully salvaged with retrograde ureteral catheterization. Time to normal temperature was 2.3 days in the percutaneous nephrostomy and 2.6 in the retrograde ureteral catheterization group, and time to normal white blood count was 2 days in the percutaneous nephrostomy and 1.7 days in the retrograde ureteral catheterization group (p not significant). Length of stay was 4.5 days in the percutaneous nephrostomy group compared with 3.2 days in the retrograde ureteral catheterization group (p not significant). Cost analysis revealed that retrograde ureteral catheterization was twice as costly as percutaneous nephrostomy. CONCLUSIONS: Retrograde ureteral catheterization and percutaneous nephrostomy effectively relieve obstruction and infection due to ureteral calculi. Neither modality demonstrated superiority in promoting a more rapid recovery after drainage. Percutaneous nephrostomy is less costly than retrograde ureteral catheterization. The decision of which mode of drainage to use may be based on logistical factors, surgeon preference and stone characteristics. PMID- 9751332 TI - Retroperitoneoscopy: experience with 200 cases. AB - PURPOSE: A retroperitoneal access is commonly used for open urological procedures. Since the introduction of the balloon dissecting technique by Gaur this anatomical route has also been used for laparoscopic surgery. We present our experience with retroperitoneoscopy in 200 cases. MATERIALS AND METHODS: From December 1992 to October 1997 a total of 200 retroperitoneoscopic procedures were performed in 197 patients 4 to 82 years old, comprising 78 nephrectomies, 50 renal cyst resections, 14 nephropexies, 11 ureterolyses, 8 retroperitoneal lymph node dissections, 8 renal biopsies, 6 adrenalectomies, 6 heminephrectomies, 6 pyeloplasties, 5 ureterolithotomies, 6 ureterocutaneostomies and 2 others. Of the patients 38 (19%) and 22 (11%) had undergone previous abdominal surgery, and kidney and ureter operations, respectively. Dissection of the retroperitoneal space was enabled by the use of a balloon catheter in 14, balloon trocar system in 93 and finger dissection technique in 93 cases. RESULTS: We classified 76 procedures (38%) as simple (renal biopsy, renal cyst resections, ureterocutaneostomy), 102 (51%) as difficult (adrenalectomy, nephrectomy, nephropexy) and 22 (11%) as very difficult (pyeloplasty, heminephrectomy, lymphadenectomy). There was a significant learning curve during the first 50 cases reflected by longer operating time, and higher complication, conversion to open surgery and open reintervention rates (14, 10 and 6%, respectively). In addition to the learning curve, mean operating time depended on the difficulty of the procedure, averaging 45 to 100 minutes for a simple, 95 to 185 for a difficult and 185 to 240 for a very difficult retroperitoneoscopy. In the last 50 cases the complication, conversion and reintervention rates (2, 4 and 2%, respectively) were acceptable for routine clinical application. CONCLUSIONS: After experience with more than 200 cases of retroperitoneoscopy the access technique has been significantly simplified. The procedure is standardized, safe and reproducible. PMID- 9751333 TI - Ureteroileal anastomotic strictures: an innovative approach with metallic stents. AB - PURPOSE: We report our experience with the use of self-expandable metallic stents to bypass anastomotic strictures after ureteroileal urinary diversion. MATERIALS AND METHODS: We evaluated 3 men and 1 woman with invasive bladder carcinoma who underwent radical cystectomy and ileal conduit urinary diversion. Ureteroenteric anastomotic strictures developed after a mean of 16 months. Self-expandable metallic stents were successfully placed (bilaterally in 2) comprising 6 stented ureters that bypassed strictures. Mean patient age was 64 years and mean followup was 12 months. RESULTS: No restenosis was observed in 3 patients during followup. The stricture recurred 1 month after stent placement in the remaining patient and additional intervention was necessary, consisting of placement of a totally coaxial overlapping metal stent. No sepsis or other complication was observed. One patient died of metastatic disease 12 months after stent placement. CONCLUSIONS: We propose the use of metal stents as an adequate, safe and effective alternative treatment for anastomotic strictures after ureteroileal diversion. PMID- 9751334 TI - Milking action: a new functional concept of a different orthotopic neobladder: 4 year followup. AB - PURPOSE: We demonstrate how the combined use of detubularized and remodeled intestine with intact cecum in the construction of an orthotopic colonic neobladder determines different functioning. MATERIALS AND METHODS: Since February 1993, 11 men who underwent radical cystectomy due to invasive bladder carcinoma have received a new bladder substitute consisting of an upper component of ascending colon and a detubularized and remodeled right half of transverse colon, and a lower component with intact cecum. During postoperative years 1 and 4 all patients were evaluated with urodynamics and cystography. RESULTS: The detubularized upper component of the neobladder acts as a large capacity, low pressure filling reservoir, while the intact cecum with its haustral contractions (inverted milking action) contributes as an additional continence mechanism. The mass contractions (milking action) with abdominal wall tension actively collaborate to evacuate the reservoir completely. CONCLUSIONS: This new structural concept of a neobladder constructed from detubularized and intact intestine has a different functional behavior than neobladders described in literature. This neobladder enables complete evacuation and total continence in the immediate postoperative period. PMID- 9751335 TI - Malone antegrade continence enema for adults with neurogenic bowel disease. AB - PURPOSE: We describe the outcomes of adults with neurogenic bowel disease who underwent a Malone antegrade continence enema procedure with or without concomitant urinary diversion. MATERIALS AND METHODS: Consecutive adult patients with neurogenic bowel disease who underwent an antegrade continence enema procedure (continent catheterizable appendicocecostomy for fecal impaction) were retrospectively reviewed. RESULTS: Of the 7 patients who underwent an antegrade continence enema synchronous urinary procedure (ileal conduit, augmentation ileocystoplasty with continent catheterizable abdominal stoma or augmentation ileocystoplasty) was also performed in 6. Mean patient age was 32 years and mean followup was 11 months. Of the 7 patients 6 who self-administered antegrade continence enemas regularly were continent of stool per rectum and appendicocecostomy, using the appendicocecostomy as the portal for antegrade enemas. All 6 compliant patients reported decreased toileting time and improved quality of life. Preoperative autonomic dysreflexia resolved postoperatively in 3 patients. All urinary tracts were stable. In 4 patients 5 complications occurred, including antegrade continence enema stomal stenosis requiring appendicocutaneous revision (1), antegrade continence enema stomal stenosis requiring dilation (1), superficial wound infection (1), small bowel obstruction requiring lysis of adhesions (1) and urinary incontinence (1 who underwent continent urinary diversion). CONCLUSIONS: Patients with neurogenic bladder and bowel disease may benefit from antegrade continence enema performed synchronously with a urinary procedure. Antegrade continence enema may be indicated alone for neurogenic bowel. Patient selection is important. PMID- 9751336 TI - Increased urinary hyaluronic acid and interstitial cystitis. AB - PURPOSE: We compared urinary levels of hyaluronic acid in patients who met the National Institute for Diabetes, and Digestive and Kidney Diseases criteria for interstitial cystitis and in age matched healthy female controls. MATERIALS AND METHODS: Urinary hyaluronic acid was measured by solid phase radiometric assay using hyaluronic acid binding protein. Hyaluronic acid and symptom scores were compared in interstitial cystitis patients who gave multiple urine samples during treatment. Since hyaluronic acid changed with treatment in some patients, 17 samples from untreated interstitial cystitis patients were selected and compared with 17 control samples. RESULTS: Mean plus or minus standard deviation urinary hyaluronic acid concentrations were similar in the 2 groups (interstitial cystitis group 574 +/- 496, controls 512 +/- 324 ng./ml., p = 0.77). When normalized to creatinine urinary hyaluronic acid was significantly higher in interstitial cystitis patients (interstitial cystitis group 674 +/- 220, controls 446 +/- 220 ng./mg. creatinine, p = 0.0019). Urinary creatinine concentrations did not differ significantly (interstitial cystitis group 842 +/- 715, controls 1,162 +/- 516 mg./l., p = 0.12). CONCLUSIONS: Urinary hyaluronic acid was higher in interstitial cystitis patients than healthy controls. Since bladder hyaluronic acid is below the epithelium, this finding may indicate leakage across the epithelium into the urine in interstitial cystitis patients. PMID- 9751337 TI - Relationship among cystectomy, microvessel density and prognosis in stage T1 transitional cell carcinoma of the bladder. AB - PURPOSE: The selection of therapy for stage T1 bladder cancer is controversial, and reliable biomarkers that identify patients likely to require cystectomy for local disease control have not been established. We evaluated our experience with T1 bladder cancer to determine whether early cystectomy improves prognosis, and whether microvessel density has prognostic value for T1 lesions and could be used for patient selection. MATERIALS AND METHODS: We retrospectively reviewed the records of 88 patients with T1 transitional cell carcinoma of the bladder. Patient outcome was correlated with therapeutic intervention. Paraffin embedded tissue from 54 patients was available for factor VIII immunohistochemical staining for microvessel density quantification. RESULTS: Median followup was 48 months (range 12 to 239). Of the patients 34% had no tumor recurrence. The rates of recurrence only and progression to higher stage disease were 41 and 25%, respectively. The survival of patients in whom disease progressed was diminished (p = 0.0002). Grade did not predict recurrence or progression nor did cystectomy provide a survival advantage. Microvessel density did not correlate with recurrence or progression. CONCLUSIONS: Patients with T1 bladder cancer have a high risk of recurrence and progression. Tumor progression has a significant negative impact on survival. Neither grade nor early tumor recurrence predicted disease progression. Because early cystectomy did not improve patient outcome, we suggest reserving cystectomy for patients with progression or disease refractory to local therapy. Microvessel density is not a prognostic marker for T1 bladder cancer and has no value in selecting patients with T1 disease for cystectomy. PMID- 9751338 TI - Predictive value of p53 and pRb immunostaining in locally advanced bladder cancer treated with cystectomy. AB - PURPOSE: We elucidate the association between altered immunostaining for retinoblastoma gene protein (pRb) and p53 nuclear proteins, and cancer specific death in patients treated with cystectomy for locally advanced bladder cancer. MATERIALS AND METHODS: The hospital records of 173 patients treated with cystectomy for advanced urothelial bladder cancer between 1967 and 1992 were retrospectively reviewed. Representative biopsies obtained before treatment were sectioned and stained using the standard immunohistochemical technique with antibody DO-7 (p53) and antibody PMG3-245 (pRb). A tumor was considered to have an altered p53 expression if 20% or more of tumor cells exhibited nuclear staining. Similarly, if no tumor cell had nuclear immunostaining the tumor was considered to have an altered pRb expression. RESULTS: An altered expression was observed for p53 in 98 tumors (57%) and for pRb in 60 (35%). In a proportional hazards analysis no association was found between an altered expression of pRb or p53 and cancer specific death. This finding was also true in another analysis when the results of immunostaining for pRb and p53 were combined. CONCLUSIONS: An altered expression for pRb and/or p53 was not correlated to cancer specific death. Thus, these parameters could not be used as predictors of treatment outcome after cystectomy for locally advanced bladder cancer. PMID- 9751339 TI - Management of female urethral diverticula: a new classification. AB - PURPOSE: Symptomatic female urethral diverticula may be managed by a number of operative techniques. However, to avoid persistent or recurrent diverticula definitive therapy requires analysis of the type and nature of the diverticulum. We propose a simple classification system for the management of female urethral diverticula. MATERIALS AND METHODS: We reviewed 18 cases of urethral diverticulectomy performed at our institution in the last 5 years. Half of the patients had been treated previously elsewhere and presented with recurring or persistent symptoms. In many cases we found a pseudodiverticulum, that is a mucosal herniation through a periurethral fascial defect. We describe our clinical distinction of a true versus pseudodiverticulum. Of 7 women with symptoms of incontinence video urodynamics demonstrated stress urinary incontinence in 4 who underwent diverticulectomy and placement of a fascial sling concurrently. RESULTS: Of 18 patients 16 were cured and 2 had persistent incontinence related to loose sling placement. Revision of the slings solved these problems. No serious complications were noted. CONCLUSIONS: Preoperative radiographic imaging helps to delineate diverticulum anatomy. Our preoperative classifications correlated well with operative findings. With meticulous excision and repair of the periurethral fascia definitive cure was achieved with a single operation. Urodynamic assessment proved crucial in achieving a successful outcome in patients with preexisting incontinence. Contrary to opinion, simultaneous placement of a sling did not lead to retropubic infection or transvaginal erosion. The placement of a sling in 4 patients achieved lasting successful repair and continence. PMID- 9751340 TI - In situ anatomical study of the male urethral sphincteric complex: relevance to continence preservation following major pelvic surgery. AB - PURPOSE: We describe a correlative gross anatomical and histological study of the human male urethral sphincteric complex using methods that delineate skeletal, muscular and fascial components. MATERIALS AND METHODS: Pelves of 6 fresh frozen male cadavers were sectioned as 4 mm. tissue blocks in planes sagittal and perpendicular to the axis of the prostatomembranous urethra from the bladder neck to the bulb of the corpus spongiosum. Sections were photographed and prepared in situ for histological staining (hematoxylin and eosin, Masson's trichrome and phosphotungstic acid hematoxylin). RESULTS: The structure of the male urethral sphincteric complex was demonstrated to include the cylindrical rhabdosphincter surrounding the prostatomembranous urethra and a fascial framework, principally consisting of the ventral subpubic fascia and medial fascia of the levator ani musculature. The histological appearance of the rhabdosphincter at its dorsal aspect suggested a suburethral musculofascial plate. Rhabdosphincteric muscle fibers were oriented in vertical and ventrolateral directions with attachments to the subpubic fascia and the medial fascia of the levator ani. CONCLUSIONS: The structural components and their relationships suggest mechanisms whereby the complex is suspended and stabilized within the deep pelvis, and achieves urethral closure. Our study furthers an understanding of the anatomical basis for male urinary continence and micturition, and is expected to have primary importance in the effort to preserve urinary function following major pelvic surgery. PMID- 9751341 TI - Dorsal onlay graft urethroplasty using penile skin or buccal mucosa in adult bulbourethral strictures. AB - PURPOSE: Preputial skin graft is used routinely for urethral reconstruction in patients with stricture disease. Alternative donor sites include extrapenile skin, bladder mucosa and buccal mucosa. Recently buccal mucosa graft has been suggested when local epithelial tissue is not available. We describe our experience with 37 patients undergoing 1-stage correction of bulbar urethral stricture using a penile skin (31) or buccal mucosa (6) graft. MATERIALS AND METHODS: In 37 patients with bulbar urethral strictures a nontubularized dorsal onlay graft was used for urethral reconstruction. A preputial skin graft was used in 31 patients and a buccal mucosa graft in 6 with a paucity of local skin. Buccal mucosa graft length ranged from 2.5 to 5 cm. (average 4) and preputial skin graft was 2.5 to 12 cm. long (average 4.7). A dorsal approach to the urethral lumen was used in all patients who underwent onlay graft urethroplasty. RESULTS: Mean followup was 21.5 months for all 37 patients, 23 months for 31 treated with preputial skin graft and 13.5 months for 6 treated with buccal mucosa graft. The clinical outcomes were considered a failure anytime postoperative instrumentation was needed, including dilatation. In the series 34 cases (92%) were classified as a success and 3 (8%) as failure. CONCLUSIONS: Onlay graft urethroplasty provided excellent results in 92% of adults with bulbourethral stricture. The dorsal approach to the urethra allowed the use of foreskin or buccal mucosa graft for reconstruction of the adequate urethral lumen. PMID- 9751342 TI - Experience with lyophilized human dura mater for urethral strictures. AB - PURPOSE: We report our results using lyophilized human dura mater for urethral strictures. MATERIALS AND METHODS: The results of 131 urethroplasties using lyophilized human dura mater grafts were reviewed. The etiology of the stricture was unknown in most cases and 44.3% had not been previously treated. Mean stricture length was 10 cm. A dura mater patch was used in 124 cases and tubular graft in 7. RESULTS: Average followup was 56.6 months. The overall complication rate during the first month was 16.3%. At 6 months the graft failed in 2 cases and 25.2% had stricture recurrence. Good results were obtained in the longer term in 41.1% of cases, with a high recurrence rate and malignancy in 4 cases (3.2%) which required penectomy. CONCLUSIONS: Despite being a biologically well tolerated material, lyophilized human dura mater is not good for urethral strictures because of the high number of relapses and possibility of malignancy. PMID- 9751343 TI - Pubovaginal fascial sling for all types of stress urinary incontinence: long-term analysis. AB - PURPOSE: There is a lack of consensus regarding indications and long-term efficacy of the many surgical techniques for treating stress incontinence. Historically pubovaginal sling has been reserved for cases of intrinsic sphincter deficiency or prior surgical failure. Transvaginal needle and retropubic suspensions have been used mainly for sphincteric incontinence unassociated with intrinsic sphincter deficiency. We report the long-term results of pubovaginal sling for all types of stress incontinence. MATERIALS AND METHODS: A total of 251 consecutive women with all types of stress incontinence who underwent pubovaginal fascial sling by a single surgeon were retrospectively and prospectively reviewed. Patients were evaluated preoperatively with history, physical examination, standardized symptom questionnaire, voiding diary, pad test, uroflow, post-void residual urine, video urodynamics and cystoscopy. Postoperatively women with at least 1-year followup were assessed by an independent third party (J. R.) who had no prior knowledge of them, and who recorded the parameters of the questionnaire, examination with a full bladder, voiding diary, pad test, uroflow and post-void residual urine. RESULTS: Overall stress incontinence was cured or improved in 92% of the patients with at least 1 year followup (median 3.1 years, range 1 to 15). The majority of patients with postoperative incontinence had de novo (3%) or persistent (23%) urge incontinence. Permanent urinary retention developed in 4 patients (2%). CONCLUSIONS: Fascial pubovaginal sling is an effective treatment for all types of stress incontinence with acceptable long-term efficacy. PMID- 9751344 TI - The etiology of post-radical prostatectomy incontinence and correlation of symptoms with urodynamic findings. AB - PURPOSE: We evaluated men with post-radical prostatectomy incontinence to determine the incidence of intrinsic sphincter deficiency and bladder dysfunction, and the contribution of each to incontinence. In addition, we determined if subjective symptoms of stress urinary incontinence and urge incontinence correlated with urodynamic findings of intrinsic sphincter deficiency and bladder dysfunction, respectively. MATERIALS AND METHODS: A total of 60 consecutive patients (mean age 64.8 years) were prospectively evaluated with multichannel video urodynamics. All patients were evaluated at least 6 months postoperatively and had achieved a stable level of continence. Patients characterized incontinence as stress or urge related, and stress urinary incontinence was graded from 0 to 3. Intrinsic sphincter deficiency was defined as incontinence associated with increased intraabdominal pressure and was further assessed by Valsalva's leak point pressure. Bladder dysfunction included urodynamic findings of detrusor instability or decreased compliance. RESULTS: Intrinsic sphincter deficiency was demonstrated in 54 patients (90%). Some component of bladder dysfunction was seen in 27 patients (45%), including detrusor instability in 24 and decreased compliance in 3, but incontinence was actually a result of bladder dysfunction in only 16 (27%). Incontinence was due to intrinsic sphincter deficiency alone in 40 patients (67%), intrinsic sphincter deficiency and bladder dysfunction in 14 (23%), and bladder dysfunction alone in only 2 (3%). Incontinence was not demonstrated on video urodynamics in 4 patients (7%). Of the 57 men who complained of stress urinary incontinence 54 demonstrated intrinsic sphincter deficiency for a positive predictive value of 95%. The 3 patients without stress urinary incontinence did not demonstrate intrinsic sphincter deficiency for a negative predictive value of 100%. Positive and negative predictive values for urge incontinence were 44 and 81%, respectively. CONCLUSIONS: Incontinence after radical prostatectomy is associated with intrinsic sphincter deficiency in the overwhelming majority of patients. Bladder dysfunction rarely is an isolated cause. When present on urodynamic tests bladder dysfunction may not always be a significant contributor to incontinence. The symptom of stress urinary incontinence (or its absence) accurately predicts the finding (or absence) of intrinsic sphincter deficiency on urodynamics. Urge incontinence is not as reliable in predicting incontinence due to bladder dysfunction. PMID- 9751345 TI - Hemodynamic effects of transurethral alprostadil measured by color duplex ultrasonography in men with erectile dysfunction. AB - PURPOSE: We evaluated the hemodynamic effects of transurethral alprostadil in 21 patients with erectile dysfunction using color duplex ultrasonography. MATERIALS AND METHODS: Penile arterial diameter, peak flow velocity and end diastolic velocity were compared following intraurethral administration of 500 microg. alprostadil and intracavernosal injection of 10 microg. alprostadil. RESULTS: A dose of 500 microg. transurethral alprostadil resulted in significant increases in corporeal blood flow comparable to those achieved with intracavernosal injection of 10 microg. alprostadil as measured by duplex ultrasonography in men with erectile dysfunction. Transurethral alprostadil resulted in statistically significant increases in arterial diameter and peak flow velocity comparable to those achieved with intracavernosal injection. End diastolic velocities were higher after transurethral alprostadil than intracavernosal injections. Color ultrasonography following transurethral alprostadil showed arterial and venous hyperemia of the corpus spongiosum and corpora cavernosa. Furthermore, color ultrasonography revealed communicating vessels between the corpus spongiosum and corpora cavernosa following administration of transurethral alprostadil. CONCLUSIONS: The visualization of communicating vessels between the corpus spongiosum and corpora cavernosa after transurethral alprostadil suggests local mechanisms of drug transfer from one to the other. In addition to potential clinical benefits, transurethral alprostadil may be useful to visualize the vascular anatomy of the penis and to test for patient responsiveness to local vasoactive agents. PMID- 9751346 TI - Efficacy and safety of transurethral alprostadil in patients with erectile dysfunction following radical prostatectomy. AB - PURPOSE: A retrospective analysis of the MUSE clinical trial was performed to evaluate the efficacy and safety of transurethral alprostadil in patients with erectile dysfunction after radical prostatectomy. MATERIALS AND METHODS: Patients received doses of transurethral alprostadil in the clinic and those for whom a suitable dose was determined were treated at home with active drug or placebo for 3 months. Patients had undergone radical prostatectomy no less than 3 months before study entry. RESULTS: Of the 384 patients in whom radical prostatectomy was identified as a cause of erectile dysfunction 70.3% had an erection believed sufficient for intercourse in the clinic and 57.1% on active medication had sexual intercourse at least once at home. The product of clinic and home success rates (70.3 x 57.1%) was an overall success rate (the likelihood of active treatment to lead to intercourse at home) of 40.1%. The frequency of most adverse effects of radical prostatectomy was comparable to that of other organic etiologies of erectile dysfunction (1,127 patients). The percentage of patients with hypotension in the clinic was lower after radical prostatectomy compared to other erectile dysfunction etiologies (0.8 versus 4.2%, p < 0.001) but the percentage of patients with urethral pain/burning was higher (18.3 versus 10.4%, p = 0.027). No urinary tract infection, fibrosis or priapism occurred in the post radical prostatectomy patients. CONCLUSIONS: Transurethral alprostadil is a well tolerated and efficacious method of treating erectile dysfunction after radical prostatectomy, although psychological changes associated with cancer and surgery may limit home response. The severe neurovascular deficit associated with prostatectomy neither limits the efficacy of transurethral alprostadil nor increases the risks. PMID- 9751347 TI - Are boxer shorts really better? A critical analysis of the role of underwear type in male subfertility. AB - PURPOSE: Elevation of testicular temperature may result in arrest of spermatogenesis, abnormal semen parameters and sterility. It has been proposed that brief style underwear may produce scrotal hyperthermia and lead to clinical subfertility. Although this idea is regarded as dogma by many in the lay community and the changing of underwear type is a therapy frequently recommended by medical practitioners, there is a paucity of data measuring scrotal temperature as a function of underwear type. MATERIALS AND METHODS: Scrotal, core and skin temperatures were measured in 97 consecutive men presenting for evaluation of primary clinical subfertility. These cases were categorized by underwear type to boxer or brief group. Semen analyses were obtained in all patients. Individuals from each group were compared to ascertain differences in temperature when wearing and not wearing underwear. Baseline semen parameters also were compared. In 14 subjects (crossover group) underwear type was changed to the alternative type and scrotal temperature measurements were repeated. Literature regarding underwear type, testicular temperature and/or fertility was reviewed and critically analyzed. RESULTS: Mean scrotal temperature plus or minus standard deviation was 33.8 +/- 0.8 C and 33.6 +/- 1.1 C in the boxer and brief group, respectively. There were no significant temperature differences between the groups. Differential temperatures comparing core to scrotal temperature and semen parameters also were not significantly different. These observations remained constant in the crossover group. CONCLUSIONS: The hyperthermic effect of brief style underwear has been exaggerated. In our study there was no difference in scrotal temperature depending on underwear type. It is unlikely that underwear type has a significant effect on male fertility. Routinely advising infertility patients to wear boxer shorts cannot be supported by available scientific evidence. PMID- 9751348 TI - Image analysis assessment of testicular touch preparation cytologies effectively quantifies human spermatogenesis. AB - PURPOSE: We have recently demonstrated that computer assisted image analysis of paraffin embedded testicular tissue based on deoxyribonucleic acid content and morphology characteristics is an effective method for the quantitative assessment of spermatogenesis. We assess the use of testicular touch preparation image analysis as a technique for quantification of spermatogenesis. MATERIALS AND METHODS: Air dried, touch imprints of testicular tissue from obstructed azoospermic and severely oligozoospermic patients were obtained at the time of biopsy. Image analysis using a filter based on deoxyribonucleic acid content and cellular morphological characteristics was performed on Feulgen stained touch preparation imprints as well as paraffin embedded sections. RESULTS: Image analysis of 52 testicular touch preparations from 48 azoospermic or severely oligozoospermic men revealed significant differences (p < 0.05) in the percentages of spermatid and spermatozoa, and 2N and 4N cells among seminiferous tubules exhibiting the 5 diagnostic categories of obstruction with normal spermatogenesis, maturation arrest at the spermatocyte stage, maturation arrest at spermatid stage, hypospermatogenesis and Sertoli cell only. Similar differences were observed in the image analysis data of the corresponding paraffin embedded testicular sections. CONCLUSIONS: Computer assisted image analysis of testicular touch preparation is an effective quantitative method of spermatogenesis evaluation. PMID- 9751349 TI - Boxers and biopsies--separating fashion from fact in male infertility. PMID- 9751350 TI - Varicocelectomy improves sperm strict morphology and motility. AB - PURPOSE: We prospectively examined the effect of varicocelectomy on standard semen parameters and Kruger strict morphology, including site of specific sperm defect. MATERIALS AND METHODS: Kruger strict morphology and routine semen analysis were performed in a blinded fashion before and a minimum of 4 months after varicocelectomy in 61 subfertile men with a primary diagnosis of varicocele. RESULTS: Sperm motility, total number of motile sperm, and percentage and total number of sperm with normal strict morphology were significantly increased after varicocele repair. Average density was also increased, while volume and forward progression were unchanged. The percentage of normal head strict morphology was significantly improved, whereas tail and neck morphology did not reflect similar changes. CONCLUSIONS: Varicocele repair in subfertile men improves Kruger strict morphology. In addition, motility and total motile sperm are significantly improved after repair. Since the stoichiometry of the sperm head is crucial to egg and sperm interaction, this decrease in head defects may be partially responsible for the increase in pregnancy rates after varicocele repair. PMID- 9751351 TI - Vasectomy reversal associated with increased reactive oxygen species production by seminal fluid leukocytes and sperm. AB - PURPOSE: Reactive oxygen species, which are primarily produced by leukocytes, are generally detrimental to sperm. High reactive oxygen species levels are found in men with abnormal sperm function. Since men often have poor sperm characteristics and infertility after vasectomy reversal, we compared reactive oxygen species in seminal cells of men after vasovasostomy to those of fertile men to determine if reactive oxygen species were elevated in the former group. MATERIALS AND METHODS: We studied semen samples of men with proved fertility (39) and those with previously proved fertility who had undergone vasectomy reversal (45). The presence of leukocytes was determined by Bryan-Leishman staining. Reactive oxygen species endogenous activity was monitored by luminol dependent chemiluminescence in washed cells, including all cells in the semen, and Percoll density gradient purified sperm. RESULTS: After vasovasostomy men had significantly lower sperm concentration, motility and computerized motility measurements than fertile men. Mean reactive oxygen species in washed seminal cells after vasovasostomy was 684 relative light units per second compared to 49 for fertile controls (p < 0.0001). Density gradient purified sperm had 53 and 0.64 relative light units per second, respectively (p < 0.0001). When men with leukocytospermia were excluded from analysis, differences between the groups remained, although 9 times more reactive oxygen species were detected in men after vasectomy reversal with than those without leukocytes in semen. CONCLUSIONS: Higher levels of reactive oxygen species are found in washed seminal cells and purified sperm after vasectomy reversal than in those of fertile men. Although leukocytes are probably a significant source of reactive oxygen species in these groups, they may not account for all of the increased reactive oxygen species after vasovasostomy. Low motility after vasectomy reversal may be related to the detrimental effects of reactive oxygen species produced by leukocytes or sperm, even in men without clinical leukocytospermia. PMID- 9751352 TI - Bleomycin associated pulmonary toxicity: is perioperative oxygen restriction necessary? AB - PURPOSE: We delineate predictive factors of pulmonary morbidity in patients who receive combination chemotherapy with bleomycin and undergo surgical resection of residual disease, and establish updated guidelines for perioperative management. MATERIALS AND METHODS: A total of 77 patients with high volume stage II to IV nonseminomatous germ cell tumors underwent 97 major surgical procedures a mean of 6.4 months following high dose combination chemotherapy, including bleomycin (mean 437.5 units per 8.2 courses), between 1988 and 1995 at the University of Texas M. D. Anderson Cancer Center. The importance of preoperative pulmonary status, anesthesia time, fraction of inspired oxygen, fluid balance, bleomycin dose, number of acute toxicity episodes, oxygen saturation problems and pulmonary symptoms was examined. Cases were divided into groups according to whether there were postoperative oxygen saturation problems (19) or not (58). RESULTS: There were no significant differences in age, weight, bleomycin dose, number of acute toxicity episodes, cardiac ejection fraction or preoperative pulmonary symptoms between the 2 groups. Restrictive spirometry patterns were seen in 26 of 74 patients (35%), only 9 of whom had postoperative oxygen saturation problems. Mean induction fractional inspired oxygen was 87% (median 100%) for an average of 56 minutes. Intraoperative fractional inspired oxygen averaged 40% for a mean duration of 8.1 hours. Postoperative oxygen saturation problems, consisting of prolonged intubation, pulmonary edema, dyspnea, tachypnea or desaturation requiring diuresis, occurred in 19 patients (25%). Surgery/anesthesia time, amount of blood transfused, estimated blood loss, fluid balance, type of fluid given (all p < 0.0001) and preoperative forced vital capacity (p = 0.012) were significant predictors of postoperative oxygen saturation problems on univariate analysis. On multivariate analysis only the amount of blood transfused, preoperative forced vital capacity and surgical time in descending order remained significant. Maintained intraoperative fractional inspired oxygen was not significant on either analysis. There were no deaths. CONCLUSIONS: Perioperative oxygen restriction in patients treated with bleomycin is not necessary. Intravenous fluid management, including transfusion, appears to be the most significant factor affecting postoperative pulmonary morbidity and overall clinical outcome. In addition, post-chemotherapy forced vital capacity and operative time are significant predictive factors of procedure related pulmonary morbidity. PMID- 9751353 TI - Intratubular germ cell neoplasia of the contralateral testis in testicular cancer: defining a high risk group. AB - PURPOSE: We define a group of testis cancer patients who are at high risk for carcinoma in situ of the contralateral testis and, therefore, a second germ cell tumor. MATERIALS AND METHODS: The histology was reviewed in 186 testis cancer patients who underwent contralateral testicular biopsy either because of a history of testicular maldescent or an atrophic contralateral testis (defined as a volume of 12 ml. or less). Testicular volume, semen analysis, serum gonadotropin levels, serum testosterone and estradiol levels were assessed in the majority of patients. RESULTS: Univariate analyses identified contralateral testicular atrophy, low sperm density, young age at presentation and low Johnsen score as factors associated with increased risk of a positive biopsy. A history of maldescent in the absence of atrophy was associated with carcinoma in situ prevalence of only 4%. Multivariate analysis identified only testicular atrophy and age at presentation as independent determinants of a positive biopsy. Testis cancer patients with a small contralateral testis had a 20% and those presenting at age 30 years or younger had a 34% prevalence, respectively, of carcinoma in situ on contralateral testis biopsy (95% confidence interval 20 and 46%, respectively). CONCLUSIONS: Testis cancer patients with an atrophic contralateral testis who present before the age of 31 years are at high risk for carcinoma in situ of the contralateral testis and, therefore, a second germ cell tumor. It is estimated that this group comprises 6% of all testis cancer patients. We predict that a policy of performing contralateral testicular biopsy will produce positive results for carcinoma in situ in a third of these patients and will detect contralateral carcinoma in situ in approximately 40% of all testis cancer patients. PMID- 9751354 TI - The impact of medical therapy on bother due to symptoms, quality of life and global outcome, and factors predicting response. Veterans Affairs Cooperative Studies Benign Prostatic Hyperplasia Study Group. AB - PURPOSE: We determine the effect of placebo, finasteride, terazosin and a combination of drugs on bother due to symptoms, quality of life and patient perception of improvement, and identify baseline clinical factors that predict clinical response to medical therapy. MATERIALS AND METHODS: A total of 1,229 subjects with clinical benign prostatic hyperplasia (BPH) were randomized to 1 year of placebo, finasteride, terazosin or drug combination. The primary outcome measures were American Urological Association (AUA) symptom score and peak flow rate. Relevant secondary outcome measures were symptom problem score, BPH impact score and global rating of improvement. RESULTS: Group mean differences in symptom problem and BPH impact scores between the finasteride versus placebo, and terazosin versus combination groups were not statistically or clinically significant. Group mean differences in all outcome measures were highly statistically significant between the terazosin and finasteride, and combination and finasteride groups. The percentage of subjects who rated improvement as marked or moderate with placebo, finasteride, terazosin and combination was 39, 44, 61 and 65%, respectively. In the subsets of men in the placebo, finasteride, terazosin and combination groups with prostates greater than 50 cm.3 group mean decrease from baseline in AUA symptom score was -2.5, -3.6, -6 and -7, group mean increase in peak flow rate was 0.6, 2.7, 3.6 and 3.7 ml. per second, group mean decrease in symptom problem score was -2.2, - 1.9, -3.1 and -4.5, and group mean decrease in BPH impact score was -0.6, -0.3, -1.1 and -1.5, respectively. A correlational analysis failed to show a significant relationship between baseline prostate volume and treatment response to finasteride. There was a significant but weak relationship between change in AUA symptom score and peak flow rate in the finasteride and combination groups. The symptom responses with terazosin were independent of baseline peak flow rate. CONCLUSIONS: In men with clinical BPH finasteride and placebo are equally effective, while terazosin and combination are significantly more effective. In men with clinical BPH and large prostates the advantage of finasteride over placebo in terms of symptom reduction, impact on bother due to symptoms and quality of life is small at best, while the advantage of terazosin and combination over finasteride and placebo is highly significant. Baseline prostate volume was not a predictor of response to finasteride in the overall study population. On the basis of our results alpha1 blockers, such as terazosin, should be first line medical treatment for BPH. PMID- 9751355 TI - Detection of abnormal E-cadherin expression by simulated prostate biopsy. AB - PURPOSE: Sampling error is an inherent problem of prostate biopsy. Consequently the determination of whether a given carcinoma is clinically significant based on biopsy results is problematic. We assess the dimensions of sampling error and, thereby, provide insight into the potential value of prognostic markers applied to needle biopsies. MATERIALS AND METHODS: We constructed 3-dimensional computer models of 21 prostatectomy specimens, including outlines of carcinomas, regions of abnormal E-cadherin expression and individual Gleason patterns. The 6 random systematic core biopsy technique and modifications were simulated using a computer algorithm. RESULTS: In 6 of 21 cases the area of abnormal E-cadherin expression and/or high grade carcinoma was not sampled on 6 random systematic core biopsy. The areas missed were either small or inconsistently under sampled regions of the prostate. Modifying the placement of biopsy needles improved the detection of these features. In addition, percent tumor in the needle appeared to be well correlated to percent tumor in the prostate (r = 0.891, r2 = 0.642). CONCLUSIONS: To avoid underestimating the aggressiveness of prostatic carcinoma at least 6 biopsies should be taken from each patient. A more extensive sampling is probably not warranted in all patients but it may prove useful in those in whom extent of disease is unclear or whose general health makes treatment decisions difficult. A reliable estimate of tumor volume in the prostatectomy specimen can be made based on relative amount of tumor in the biopsy specimen on an individual basis. PMID- 9751356 TI - Obliterative vesicourethral strictures following radical prostatectomy for prostate cancer: reconstructive armamentarium. AB - PURPOSE: We report the reconstructive techniques used to correct obliterative vesicourethral strictures related to prostate cancer surgery. MATERIALS AND METHODS: Four men with anastomotic obliteration after radical prostatectomy underwent primary excision with end-to-end anastomosis, penile fasciocutaneous flap, free-graft urethroplasty with rectus muscle flap or anterior bladder tube with omental pedicle flap procedure. RESULTS: At mean followup of 33.8 months all patients had urethral patency but none was continent. CONCLUSIONS: Single stage reconstruction of the obliterated vesicourethral anastomosis after prostatectomy successfully restored urethral patency. No technique was applicable in all cases. Sphincteric function is likely to be compromised after the primary procedure, resulting in incontinence after successful urethral reconstruction. Subsequent artificial sphincter placement appears to be safe and helpful in restoring continence. PMID- 9751357 TI - Transient lower extremity neurapraxia associated with radical perineal prostatectomy: a complication of the exaggerated lithotomy position. AB - PURPOSE: We assess the incidence and risk factors associated with lower extremity neurapraxia following radical perineal prostatectomy. MATERIALS AND METHODS: The medical records of 111 consecutive patients undergoing radical perineal prostatectomy at Duke University Medical Center between June 1994 and June 1995 were retrospectively reviewed. Patients were interviewed by telephone to ascertain whether symptoms had resolved. RESULTS: Neurapraxia developed in 23 patients (21%). Symptomatology was variable, including sensory and motor deficits of the lower leg and foot. Although lower extremity neurapraxia occurred in a significant number of patients undergoing radical perineal prostatectomy, it appeared to resolve in most. CONCLUSIONS: Careful attention to detail when positioning the patient and limiting the time in the exaggerated lithotomy position appear to be the most critical aspects to prevent neurapraxia. PMID- 9751358 TI - Identification of patients at increased risk for prolonged urinary retention following radioactive seed implantation of the prostate. AB - PURPOSE: Urinary retention is a frequently reported complication following radioactive seed implantation of the prostate. If retention is refractory, a post implant transurethral prostatic resection may ultimately be required to relieve obstruction, leading to an increased risk of urinary incontinence. In this series the incidence of prolonged urinary retention was determined, and the effect of pretreatment and treatment related factors was analyzed to identify high risk patients. MATERIALS AND METHODS: A total of 251 patients with organ confined prostate carcinoma underwent transperineal prostate seed implantation. Of the patients 114 were implanted with 103palladium (103Pd) and 137 with 125iodine seeds. Of the patients who were implanted with 103Pd 90 received 3 months of neoadjuvant hormonal therapy. All patients had International Prostate Symptom Scores (I-PSS) recorded before implantation to assess the degree of urinary symptoms. In the patients receiving neoadjuvant hormones prostate volumes and I PSS were recorded before initiation of hormone treatment and 3 months later at the time of implant. RESULTS: Urinary retention developed in 14 patients requiring catheterization for more than 48 hours. Median time to onset was 1 day after implant. Of these patients 6 ultimately required transurethral prostatic resection to relieve urinary obstruction. No patient had urinary incontinence following implantation or transurethral prostatic resection. Multivariate analysis revealed that pretreatment I-PSS, and combined treatment with hormonal therapy and 103Pd predicted for the development of retention. Patients with I-PSS 20 or greater had a 29% risk, I-PSS 10 to 19, 11% risk and I-PSS less than 10, 2% risk of retention. Neither patient age, clinical stage, prostate specific antigen, Gleason score, use of 125I nor prostate volume was significant. A subgroup analysis of patients receiving hormonal therapy and 103Pd revealed that those with persistent urinary symptoms (I-PSS 10 or greater) following 3 months of hormonal therapy had the greatest risk of prolonged retention (37%). CONCLUSIONS: The overall risk of prolonged urinary retention following prostate implantation was low in our series. Using the I-PSS questionnaire, high risk patients can be identified before treatment. Patients with significant pretreatment urinary symptoms or persistent urinary symptoms following 3 months of hormonal therapy and then implantation with 103Pd have the greatest risk. PMID- 9751359 TI - Radical prostatectomy for prostate cancer: the perineal approach increases the risk of surgically induced positive margins and capsular incisions. AB - PURPOSE: We compare the incidence of positive surgical margins in patients who underwent perineal or retropubic radical prostatectomy for clinically localized (stage T1, T2) prostate cancer. MATERIALS AND METHODS: In this retrospective, nonrandomized study we reexamined the specimens of 94 consecutive patients who underwent radical perineal (48) or retropubic (46) prostatectomy for clinically localized prostate cancer (stage T1, T2) and with pathological stage pT2 (intracapsular), pT3A (established extracapsular extension without positive margins) or pT3B (extracapsular extension with positive margins) without lymph node involvement (N0). We assessed the presence or absence of extracapsular cancer with or without positive margins, incisions of the prostatic capsule exposing cancer (surgically induced positive margins) or benign glandular tissue. Patients were followed for 3 to 66 months (mean 25) using an ultrasensitive prostate specific antigen assay with a lower detection limit of less than 0.05 ng./ml. RESULTS: The overall incidence of positive margins in cancer tissue was 56% in the perineal and 61% in the retropubic group, and biochemical failure-free survival was 67% each. However, surgically induced positive margins in patients with organ confined disease were more frequent in the perineal than retropubic group (43 versus 29%, p < 0.05) and associated with a 37% risk of biochemical failure (prostate specific antigen greater than 0.1 ng./ml.) at mean followup. In addition, capsular incisions exposing benign tissue were more frequent in the perineal than retropubic group (90 versus 37%, p < 0.05) irrespective of pathological stage. CONCLUSIONS: Although overall positive margins and biochemical failure rates are similar or identical for the perineal and retropubic approaches for organ confined prostate cancer, the perineal approach is associated with a significantly higher risk of capsular incisions and surgically induced positive margins and, thus, a higher risk of biochemical failure. PMID- 9751360 TI - Complications of treatment for localized prostate cancer. PMID- 9751362 TI - Progression in and survival of patients with locally advanced prostate cancer (T3) treated with radical prostatectomy as monotherapy. AB - PURPOSE: We determine the progression and survival rates in patients with locally advanced prostate cancer treated with radical prostatectomy without adjuvant treatment, and investigate subgroups of patients who may not benefit from this treatment. MATERIALS AND METHODS: Radical prostatectomy was performed in 83 patients with T3 prostate cancer. The patients were divided in subgroups with T3G1 to 2 and T3G3 tumors, which were evaluated for clinical progression, local recurrence, distant metastases, biochemical progression, and overall and cancer specific survival at 5 and 10 years by Kaplan-Meier curves. The results were compared to those of 190 patients with locally confined tumors. RESULTS: At 5 and 10 years overall survival was 75 and 60%, and cancer specific survival was 85 and 72%, respectively. At 5 and 10 years clinical progression was 41 and 69%, local recurrence 18 and 44%, and distant metastases 31 and 50%, respectively. Biochemical progression at 5 years was 71%. Patients with poorly differentiated tumors showed significantly lower survival and higher progression rates compared to those with well or moderately differentiated tumors. Progression and survival in patients with T3G1-2 tumor were not significantly different from those for patients with locally confined tumors. CONCLUSIONS: Radical prostatectomy as monotherapy in patients with locally advanced nonmetastatic prostate cancer (T3) produces acceptable results in those with well or moderately differentiated tumors. The results of progression and survival are not significantly different from those in patients with locally confined prostate cancer. However, patients with poorly differentiated tumors (T3G3) have early progression and need adjuvant treatment following surgery. PMID- 9751361 TI - Limited role of radionuclide bone scintigraphy in patients with prostate specific antigen elevations after radical prostatectomy. AB - PURPOSE: Bone scintigrams of patients with increasing serum prostate specific antigen (PSA) after radical prostatectomy are only rarely positive. We identify clinical parameters that would improve our ability to select patients for this imaging study. MATERIALS AND METHODS: We reviewed all bone scintigrams done at our institution between 1991 and 1996 in patients with persistently increasing serum PSA after radical prostatectomy. What prompted the clinician to obtain the bone scintigram was trigger PSA (tPSA). The rate of increase in PSA to tPSA was measured by tPSA/time from radical prostatectomy (slope 1) and tPSA/time from last undetectable PSA (slope 2). These parameters were evaluated together with standard clinicopathological data in univariate and multivariate analyses to determine the ability to predict the bone scintigram result. RESULTS: In univariate analysis tPSA (p = 0.003), slope 1 (p = 0.005) and slope 2 (p = 0.004) were useful in predicting the bone scintigram result but pathological stage, Gleason score, preoperative PSA and time to recurrence were not. In multivariate analysis the single most useful parameter in predicting the bone scintigram result was tPSA (p = 0.01). Based on a logistic regression model the probability of a positive bone scintigram was less than 5% until tPSA increased to 40 to 45 ng./ml. CONCLUSIONS: In patients with increasing serum PSA after radical prostatectomy current serum PSA is the best predictor of the bone scintigram result. Furthermore, there is limited usefulness of bone scintigraphy until PSA increases above 30 to 40 ng./ml. PMID- 9751363 TI - Use of second treatment following definitive local therapy for prostate cancer: data from the caPSURE database. AB - PURPOSE: We compare secondary cancer treatment use in patients who underwent definitive local treatment for prostate cancer. MATERIALS AND METHODS: The rate of second cancer treatment was determined in patients who underwent radical prostatectomy (1,254), radiotherapy (499) or cryosurgery (141) using data from the CaPSURE database, a longitudinal disease registry of patients with prostate cancer. Second treatments started within 3 months after initial treatment were defined as adjuvant and those started more than 3 months were defined as nonadjuvant. Using a parametric regression model of survival analysis, second treatment rates were adjusted for differences in clinical and demographic characteristics, and duration of followup among groups. RESULTS: Of the patients 4% received a second adjuvant treatment and 17% received a second nonadjuvant treatment within 3 years of initial therapy. Adjusted rates of nonadjuvant second treatment were lowest after radical prostatectomy, and 34 and 88% higher after radiation and cryosurgery, respectively (p = 0.01). This finding was most evident in patients with pretreatment prostate specific antigen 10.0 ng./ml. or less, clinical stage T2N0M0 disease, or Gleason score 6 or less on diagnostic biopsy, and in those classified as low risk for recurrence based on a combination of these parameters (p = 0.004). CONCLUSIONS: Approximately 1 in 5 patients receive second cancer treatment within a mean of 3 years following initial local treatment for prostate cancer. Our data suggest that the likelihood of receiving second treatment was lowest in patients initially treated with radical prostatectomy. PMID- 9751364 TI - Vitamin D receptor polymorphisms and lethal prostate cancer. AB - PURPOSE: Reports in the osteoporosis literature demonstrating the increased activity of specific alleles of the vitamin D receptor and epidemiological data linking vitamin D levels with prostate cancer have stimulated research into possible associations between vitamin D receptor genotype and the development of prostate cancer. Recent studies showed that patients homozygous for a less active vitamin D receptor have a 4 to 5 times increased risk of localized prostate cancer. In 1 study this association was strongest in patients with advanced disease. To understand better the relationship between advanced disease and the vitamin D receptor we compared the vitamin D receptor genotype of 41 patients who died of prostate cancer to 41 controls with no clinical evidence of prostate cancer. MATERIALS AND METHODS: Noncancerous deoxyribonucleic acid was isolated from lethal prostate cancer and control cohorts. To determine the TaqI restriction fragment length polymorphism a 740 base pairs (bp) segment of the vitamin D receptor was amplified by PCR, digested with TaqI endonuclease and resolved on an agarose gel. Depending on the presence or absence of a TaqI restriction site at codon 352 in each allele, products were digested into 2 fragments of 495 and 245 bp (T allele) or 3 fragments of 290, 245 and 205 bp (t allele). Individuals were classified as TT, Tt or tt. To determine the size of the poly-A microsatellite repeat an approximately 410 bp fragment was amplified by polymerase chain reaction using [gamma-32P] labeled primers, resolved by gel electrophoresis and sized by autoradiography. Fragments 410 bps or greater corresponded to repeats 18 bps or greater (L allele) and fragments less than 410 bps corresponded to repeats less than 18 bps (S allele). Individuals were classified as LL, LS or SS. RESULTS: The TaqI restriction fragment length polymorphism and poly-A microsatellite repeat were in strong linkage disequilibrium, indicating that both polymorphic markers identified the same vitamin D receptor genotype in the majority of cases. Using the TaqI restriction fragment length polymorphism 33 of 41 controls (80.5%) and 35 of 41 cases (85.3%) were heterozygous (Tt) or homozygous (TT) for the less active vitamin D receptor allele, while 8 of 41 controls (19.5%) and 6 of 41 cases (14.6%) were homozygous (tt) for the more active vitamin D receptor allele. Using the poly-A microsatellite repeat 32 of 40 controls (80%) and 32 of 38 cases (84.2%) were heterozygous (LS) or homozygous (LL) for the less active vitamin D receptor allele, while 8 of 40 controls (20.0%) and 6 of 38 cases (15.8%) were homozygous (SS) for the more active vitamin D receptor allele. There was no statistically significant difference in the distribution of either marker between cases and controls. CONCLUSIONS: We failed to demonstrate an association between vitamin D receptor genotype and lethal prostate cancer. Our data do not support the hypothesis that specific vitamin D receptor genotypes are associated with an aggressive prostate cancer phenotype. Further studies that take into account cofactors important in vitamin D activity and/or a better definition of prostate cancer phenotype may explain the discrepancy between our findings and those of a previous study. PMID- 9751365 TI - Adverse effects on vasoepididymostomy outcomes for men with concomitant abnormalities in the prostate and seminal vesicle. AB - PURPOSE: Following microsurgical vasoepididymostomy as many as 85% of men have sperm in the ejaculate, yet only 30 to 50% will spontaneously father children. We examined the possibility that there may be concomitant abnormalities in the prostate and seminal vesicle, which may be associated with low pregnancy rates. MATERIALS AND METHODS: Transrectal ultrasound was performed in azoospermic men with suspected epididymal obstruction, excluding those who had undergone vasectomy, to identify abnormalities of the seminal vesicles and ejaculatory ducts. Microsurgical vasoepididymostomy was attempted in all men. RESULTS: Transrectal ultrasound revealed ejaculatory duct dilatation in 13 of 40 men (33%), although only 3 had accompanying seminal vesicle dilatation. Two men had atrophic seminal vesicles with normal ejaculatory ducts. At surgery 8 of 40 patients (20%) were deemed to have irreparable conditions. For the 27 men followed at least 6 months postoperatively patency and pregnancy rates were 75 and 22%, respectively. Mean sperm counts plus or minus standard deviation were significantly higher in men without compared to those with seminal vesicle or ejaculatory duct abnormalities (43 +/- 68 versus 5.7 +/- 6.9 x 10(6) sperm per ml., respectively), and so was the percentage of motile sperm (30 +/- 16% versus 1.2 +/- 2.2%, respectively). Pregnancy rates were also higher in men without (6 of 19, 32%) than with (0 of 8, 0%) seminal vesicle or ejaculatory duct abnormalities. CONCLUSIONS: Transrectal ultrasound detected abnormalities of the seminal vesicles and ejaculatory ducts are common in men with suspected epididymal obstruction. These abnormalities are associated with a poor outcome for vasoepididymostomy. We recommend that all men with suspected epididymal obstruction undergo transrectal ultrasound before any attempted reconstruction. PMID- 9751366 TI - Transcontinental interactive laparoscopic telesurgery between the United States and Europe. PMID- 9751367 TI - An effective technique to facilitate radiographic stone visualization with an internal stent during shock wave lithotripsy. AB - PURPOSE: We describe a simple method to assist stone localization during shock wave lithotripsy in the presence of a Double J stent. MATERIALS AND METHODS: A 4F whistle tip ureteral catheter is passed alongside a previously inserted 6F Double J stent. The tip of the ureteral stent is positioned in the lower or mid third of the ureter. Contrast material is injected through the ureteral catheter during lithotripsy to assist stone localization. RESULTS: This technique has been successful in localization of poorly opacified renal stones during lithotripsy. CONCLUSIONS: Radiolucent and poorly calcified renal stones can be easily localized during shock wave lithotripsy, despite the presence of a Double J stent. No special catheters or stents are required for this technique. PMID- 9751368 TI - An alternative ureteroileal reimplantation used in augmentation cystoplasty for neurogenic bladder with bilateral vesicoureteral reflux. AB - PURPOSE: We describe the use of a serous lined extramural tunnel for ureteral reimplantation during augmentation of a neurogenic bladder to prevent reflux. MATERIALS AND METHODS: A 46-year-old male C6 spinal cord injury patient presented with a high pressure bladder, detrusor-sphincter dyssynergia and bilateral grade II/III vesicoureteral reflux. Despite maximal anticholinergic therapy and intermittent catheterization, detrussor pressures were between 80 and 100 cm. water at volumes of 100 to 150 cc with consistent leakage between catheterizations. Preoperative ultrasound and voiding cystourethrogram demonstrated moderate bilateral hydronephrosis and a heavily trabeculated bladder. Augmentation cystoplasty with the formation of 3 cm. extramural ureteral tunnels as described by Ghoneim was performed. The serosa of the adjacent limbs of the ileal segment were opposed to form the back wall of a serosal lined tunnel. RESULTS: At 3 weeks postoperatively a cystogram demonstrated no extravasation or reflux. At 8 weeks an excretory urogram showed prompt function and excretion bilaterally with marked improvement of preoperative hydronephrosis. CONCLUSIONS: Subserosal ureteral tunnels can be used as an alternative antireflux technique during augmentation cystoplasty when ureteral reimplantation is required. Two advantages of this technique include the elimination of staples and avoidance of ischemic problems associated with an afferent intussuscepted nipple valve. PMID- 9751369 TI - Cavernous hemangioma of the adrenal gland in a patient on chronic hemodialysis. PMID- 9751370 TI - A simple means of making the differential diagnosis of ureterouterine and vesicouterine fistula. PMID- 9751371 TI - Recovery of renal function after 153 days of complete unilateral ureteral obstruction. PMID- 9751372 TI - Adenocarcinoma in an isolated ileal sleeve 30 years after periureteral wrap. PMID- 9751373 TI - Successful treatment of metastatic skin lesions with electrochemotherapy. PMID- 9751374 TI - Osteomyelitis as a complication of vesica percutaneous bladder neck suspension. PMID- 9751375 TI - Obstructive vaginal angiomyoma. PMID- 9751377 TI - Uncommon course of carcinoma of the prostate after radical prostatectomy and luteinizing hormone-releasing hormone analogue therapy. PMID- 9751376 TI - Ulcerative balanoposthitis as the initial manifestation of acute promyelocytic leukemia. PMID- 9751378 TI - Atypical true hermaphroditism with a mosaic 45,X/46,X,dic(Y) (q11.2) karyotype. PMID- 9751379 TI - Iatrogenic muscle damage after prolonged surgery discovered by radionuclide bone scanning. PMID- 9751380 TI - Re: Influence of technique of percutaneous tract creation on incidence of renal hemorrhage. PMID- 9751381 TI - Re: Editorial: Alternatives to appendix in construction of a mitrofanoff stoma. PMID- 9751382 TI - Re: Testicular descent: a proposed interaction between mullerian inhibiting substance and epidermal growth factor. PMID- 9751383 TI - Re: Bacterial infection in prostatodynia. PMID- 9751384 TI - Re: Detection of prostate specific antigen in pancreas and salivary glands: a potential impact on prostate cancer overestimation. PMID- 9751385 TI - Re: Editorial: The prostate puzzle. PMID- 9751386 TI - Re: Recurrence patterns after radical retropubic prostatectomy: clinical usefulness of prostate specific antigen doubling times and log slope prostate specific antigen. PMID- 9751387 TI - Re: Long-term outcome in patients with pTxN+ adenocarcinoma of prostate treated with radical prostatectomy and early androgen ablation. PMID- 9751388 TI - Re: Schistosomiasis of the male genital tract: transrectal sonographic findings. PMID- 9751389 TI - Re: The value of unenhanced helical computerized tomography in the management of acute flank pain. PMID- 9751390 TI - The captopril renogram: a new tool for diagnosing and predicting obstruction in childhood hydronephrosis. AB - PURPOSE: We evaluated the difference in response patterns of captopril versus standard renography for assessing hydronephrosis due to suspected ureteropelvic and ureterovesical junction obstruction. This technique may identify hydronephrotic kidneys in which normal function is maintained by vasoactive compensatory mechanisms. Sustained obstruction may cause these mechanisms to fail, and expose the kidneys to permanent functional deterioration in the long term. MATERIAL AND METHODS: We prospectively studied 15 boys and 8 girls with a mean age of 3.5 years with grades III to IV/IV hydronephrosis. Evaluations included renal sonography, standard diuretic and captopril renography, glomerular filtration rate, voiding cystography, serum creatinine, blood pressure, and urinalysis with culture and sensitivity. Obstruction was suspected at the ureteropelvic junction in 19 kidneys and at the ureterovesical junction in 9. We compared differential function values obtained by standard diuretic and captopril renography. RESULTS: We observed certain patterns in response to captopril renography, including pattern 1--unilateral decrease in hydronephrotic kidney relative function in 5 of 23 cases, 2--bilateral decreased function in 2, 3- bilateral increased function in 4 and 4--no change in function in 12 on standard renography. When half-time was more than 20 minutes on standard diuretic renography in 8 cases, captopril renography showed an ipsilateral decrease and bilateral increase in glomerular filtration rate in 4 and 1, respectively, and no change in 3. In 12 of the 23 patients (52%) there was no difference in the results of captopril and diuretic renography as well as no change in differential function on standard diuretic renography during 1 1/2 years of observation. Surgical correction was performed in 4 patients in whom half-time was greater than 20 minutes and differential function was decreased on captopril renography. CONCLUSIONS: Our preliminary study reveals that there may be a role for captopril renography for detecting renin-angiotensin system mediated compensatory mechanisms in obstructive uropathy. When such compensatory mechanisms are activated, they may be unmasked by captopril, producing 1 of 4 patterns on renography and glomerular filtration rate. Patterns may indicate different degrees of impending renal function impairment and, thus, they may become useful for determining the progression of injury, when present, and the appropriate timing of surgical intervention. PMID- 9751392 TI - A unique new model to study the effects of urinary diversion in the developing rabbit bladder. AB - PURPOSE: Little is known about the developmental effects of high urinary diversion and bladder defunctionalization in infancy. Although clinical experience shows that a poorly functional bladder may result from urinary diversion in infancy, the mechanisms of change and specific bladder wall alterations have not been well characterized. We hypothesized that cyclic filling and emptying are necessary for normal bladder development. To investigate this important question we created a new animal model. MATERIALS AND METHODS: We designed a new method of hemibladder urinary diversion in 3-week-old New Zealand white rabbits. After vertical midline bladder division half of the bladder was formed into a functional reservoir, which remained in continuity with the ipsilateral ureter and urethra. The other bladder half was defunctionalized and isolated from the urine flow by ureteral ligation. Diversion was created for 3, 7, 14 and 28 days. Urodynamic evaluation was done in the functionalized hemibladders and age matched normal rabbit bladders to test the validity of the functionalized hemibladder as an internal control. Functional and defunctionalized hemibladders as well as age matched, nonoperated normal rabbit bladders were weighed, sectioned and stained to demonstrate muscle and connective tissue components. RESULTS: In 22 of the 27 healthy rabbits (81%) good quality diverted and functional bladder specimens were obtained after diversion. Defunctionalized hemibladders grew more slowly than functionalized bladders and normal age matched control bladders. Histological staining of the bladder wall demonstrated increased connective tissue between the muscle bundles within the diverted specimens than in functional bladders. CONCLUSIONS: Our successful model of urinary diversion may be used to study the developmental and histological effects of urinary diversion in the young bladder. Bladder growth and histological appearance are altered when the stimulus of cyclic filling and emptying is removed. Further studies using this model are warranted to define fully bladder changes that result from diversion and investigate the mechanism of the observed changes. PMID- 9751391 TI - Conservative surgical management of bilateral Wilms tumor: results of the United Kingdom Children's Cancer Study Group. AB - PURPOSE: Bilateral Wilms tumor presents the clinician with a treatment dilemma. Since 1980 most centers of the United Kingdom Children's Cancer Study Group have used a conservative surgical approach with initial biopsy followed by chemotherapy and delayed surgical resection. We assess the outcome of this treatment approach in terms of survival, and preservation of renal mass and function. MATERIALS AND METHODS: We retrospectively analyzed the records of 71 children with bilateral Wilms tumor diagnosed between 1980 to 1995 at 17 United Kingdom Children's Cancer Study Group centers. In 57 patients conservative surgical treatment with initial biopsy was followed by chemotherapy and delayed tumor resection, while 13 underwent initial surgical resection followed by chemotherapy. One patient was excluded from study because the lesion in 1 kidney proved to be a benign cyst. Mean followup was 6 years (range 1 to 15). The percentage of renal tissue involved with tumor and preserved was estimated, and renal function at the last followup was recorded. RESULTS: Overall survival was 69% with similar survival in the conservatively treated and initial surgical resection groups. At the last followup renal function was normal in 80% of the patients in each group. Mean preserved renal mass was 45 and 35% in the conservatively treated and initial resection groups, respectively, with a trend toward better preservation in those treated conservatively. Bilateral Wilms tumor with an unfavorable histology was associated with a poor prognosis. CONCLUSIONS: Conservative surgical treatment of favorable histology bilateral Wilms tumor may improve the preservation of renal mass and function without impairing patient survival. PMID- 9751393 TI - Noninvasive evaluation of bladder compliance in children using ultrasound estimated bladder weight. AB - PURPOSE: In healthy children as well as those with urinary disturbance we determined ultrasound estimated bladder weight with the aim of revealing its possible usefulness as a measure of bladder compliance. MATERIALS AND METHODS: We measured ultrasound estimated bladder weight in 71 healthy children with a mean age of 10.3 years, and determined a standard value. A total of 82 patients with a mean age of 9.6 years with urinary disturbance also underwent ultrasound estimated bladder weight measurement as well as conventional urological examinations, including filling cystometry. RESULTS: In healthy children ultrasound estimated bladder weight increased with age, showing a significant linear correlation (r = 0.80, p < 0.0001). Using the formula for linear correlation, 0.86 x patient age + 6.9 gm., we obtained an age matched estimated weight. In 82 patients the percent deviation of the estimate from age matched values was calculated using the formula, (measured ultrasound estimated bladder weight -age matched ultrasound estimated bladder weight)/age matched ultrasound estimated bladder weight x 100, and then correlated with bladder compliance. In 75 of 77 patients (97%) with compliance of 10 ml./cm. water or more the estimate was within 100% deviation. In contrast, 4 of 5 patients (80%) with a low compliant bladder (less than 10 ml./cm. water) had an ultrasound estimated bladder weight greater than 100% deviation. When the estimate was within 100% deviation, all but 1 patient (75 of 76, 98.7%) had compliance of 10 ml./cm. water or more compared to 33.3% (2 of 6) of those with an estimate greater than 100% deviation. As a result, with the use of a cutoff value of 100% deviation ultrasound estimated bladder weight predicted a low compliant bladder with a diagnostic accuracy as high as 96.3% (79 of 82 cases). CONCLUSIONS: Ultrasound estimated bladder weight may be used to evaluate bladder compliance in children. It seems to be a suitable noninvasive urodynamic test in children with suspected urodynamic abnormalities. PMID- 9751394 TI - Premature urachal closure induces hydroureteronephrosis in male fetuses. AB - PURPOSE: Congenital hydronephrosis is more common in male individuals. We investigate whether an alteration in fetal bladder function induced by premature urachal closure contributes to fetal hydronephrosis, consequently explaining this male predominance. MATERIALS AND METHODS: The urachus was clipped in 8 male and 4 female ovine fetuses at 95 days of gestation (term 140 days). Subjects were sacrificed, and the urinary tract was assessed at 109 and 116 days of gestation in 3 and 1 male fetuses, respectively, and at term in 4 male and 4 female lambs. RESULTS: At 109 and 116 days of gestation 3 of the 4 male fetuses had upper tract dilatation. At term no female but 4 male lambs had hydroureteronephrosis, including some with marked pelvic dilatation and parenchymal thinning. At term mean bladder weight in the male animals with urachal clipping was 5.28 gm. (range 3.5 to 8.2), which was significantly greater than normal (p = 0.02). Bladder weight at term in the female lambs with urachal clipping was not different from normal values. Histological evaluation of the kidneys in the male lambs revealed cortical thinning and loss of medullary tissue, while the overall renal architecture was well preserved. CONCLUSIONS: Our observations indicate that normal ovine fetal urinary tract function and drainage depend on urachal function and the timing of urachal closure. Therefore, fetal hydronephrosis is associated with this alteration in bladder function but it may also be associated with other factors, such as bladder sphincter maturation or prostate development. Our experiment shows that hydroureteronephrosis develops in ovine fetuses when all bladder drainage occurs via the urethra. This condition may be an amplification of the differences in bladder outlet resistance in human fetuses, which may explain the male predominance in the various forms of hydroureteronephrosis. PMID- 9751395 TI - Bladder dysfunction: an integral part of the ectopic ureterocele complex. AB - PURPOSE: We evaluate whether bladder dysfunction is common in patients with ectopic ureterocele and, if so, whether it is an integral part of the ectopic ureterocele complex or a result of surgery. MATERIALS AND METHODS: From 1986 to 1995, 34 patients with a mean age of 10 months were treated for large or medium ectopic ureteroceles at our institution and 32 participated in postoperative followup. Bladder function was investigated by a careful history and repeat uroflowmetry, and residual urine estimation was assessed by ultrasound and cystometry. RESULTS: Of the 32 patients 19 had infrequent voiding and 3 had incontinence. Cystometric bladder capacity was increased to greater than 150% of the normal value for age in 15 of 27 patients (55%). Uroflowmetry revealed greater than 5 ml. residual urine in 15 patients (56%). Postoperatively no radiological signs of bladder neck obstruction were found. Increased bladder capacity and residual urine did not correlate with ureterocele size or location, or surgical procedure. There was no progression of bladder dysfunction with age. CONCLUSIONS: Children with ectopic ureterocele are at high risk for a high capacity bladder with incomplete emptying. This bladder dysfunction associated with ectopic ureterocele does not seem to be the result of surgery but an integral part of the disorder. PMID- 9751396 TI - Long-term adjustment issues in patients with exstrophy. AB - PURPOSE: We explored the psychological adjustment of children with bladder or cloacal exstrophy. MATERIALS AND METHODS: We assessed 29 subjects with a mean age plus or minus standard deviation of 7.8 +/- 3.97 years using age appropriate standard psychological instruments. Psychological adjustment scores in the exstrophy group were compared to the norms of the various instruments. Subjects were divided into dichotomous groups according to several medical and demographic factors. For each factor the differences between the means of the 2 groups on the outcome variables were calculated using a t test. RESULTS: Children with exstrophy perceived their appearance more positively than the norm. Older children performed more poorly than younger children in adaptive behavior, specifically in skills related to functioning in school. Children who achieved continence after age 4 years were more likely to have problems with acting out behavior. There were no differences in adjustment in boys versus girls, bladder versus cloacal exstrophy, type of continence strategy or gender reassignment versus no reassignment. CONCLUSIONS: Children with exstrophy did not have clinical psychopathology. Differences existed in adaptive and acting out behavior rather than depression or anxiety, suggesting that improved outcomes may be achieved through a focus on normal adaptation rather than on potential psychological distress. PMID- 9751397 TI - Posterior sagittal approach for management of a traumatic urethral stricture. PMID- 9751398 TI - Four-hour voiding observation in young boys with posterior urethral valves. AB - PURPOSE: We evaluated 4-hour voiding observation as a method of basic assessment of bladder dysfunction in young boys with posterior urethral valves. MATERIALS AND METHODS: Voiding pattern, including number of voids, voided and residual urine volume, and bladder capacity, was determined noninvasively in 24 boys younger than 4 years with posterior urethral valves and compared to that of healthy age matched controls. Results were then compared to those of standard cystometry. RESULTS: The number of voids was higher, voided volume was smaller and residual urine volume was higher in the posterior urethral valve group. There was no difference in voiding pattern before and after removal of the anatomical obstruction. Voided and residual urine volume, and bladder capacity were higher on standard cystometry than on voiding observation. CONCLUSIONS: Four-hour voiding observation is an easy noninvasive method that focuses on emptying difficulties and clearly detects differences in voiding patterns between boys with posterior urethral valves and healthy, nontoilet trained children. We recommend the method as a complement to standard cystometry for the diagnosis and followup of bladder dysfunction in young boys with posterior urethral valves to identify the need for treatment. PMID- 9751399 TI - Anatomical alignment for the correction of buried penis. AB - PURPOSE: We report a straightforward surgical technique for the correction and anatomical alignment of the skin in patients with various degrees of buried penis. MATERIALS AND METHODS: A combined series of 74 patients 7 months to 10 years old who were treated for buried penis at 2 institutions during a 7-year period. Patients presented with various symptoms, including balanitis, urinary tract infection, painful voiding, ballooning of the foreskin and urinary retention. In 29 patients (38%) trapped penis was due to previous circumcision. In our estimation the major anatomical defect in buried penis is an insufficient attachment of the dartos fascia and penile skin to Buck's fascia. Our technique involves making a circumferential incision of the inner preputial skin layer proximal to the corona, unfurling it from the shaft skin and leaving a coronal collar of approximately 1 cm. The annular band that usually constricts the corpora on retraction of the penile skin is incised, and the remaining proximal penile skin and dartos fascia are dissected off Buck's fascia proximally to the base of the penis. The penile dermis is sutured to the lateral aspect of the tunica albuginea at the penopubic junction and mid shaft of the penis. This technique restores normal anatomical relationships with excellent cosmetic results and negligible complications. RESULTS: At a median 5-year followup cosmesis was excellent in all cases. Two patients with micropenis who required revision responded to endocrine therapy. CONCLUSIONS: Excellent cosmetic results were obtained in all cases using this surgical technique. PMID- 9751400 TI - Primary lymphedema of the genitalia in children and adolescents. AB - PURPOSE: Congenital lymphedema is a rare disorder that may result in disfiguring edema of the male genitalia. We reviewed our experience with 5 cases to advance our understanding of this challenging problem. MATERIALS AND METHODS: Four boys with significant lymphedema underwent excision of the involved subcutaneous genital tissue and coverage with local skin flaps. Two boys in whom this approach failed later underwent complete excision of the involved subcutaneous tissue and skin, and coverage with split thickness skin grafts. The boy with minimal edema was observed. RESULTS: Two of the 4 boys who underwent subcutaneous genital tissue resection and coverage with local skin flaps are markedly improved, although 1 requires further revision. In the other 2 boys treatment failed, necessitating repeat genital tissue excision and grafting. While there have been no recurrences in the grafted areas, each patient has required additional operations to manage recurrent edema in adjacent tissues of the perineum and inguinal region, and in 1 significant contraction of the grafted skin developed. Mild genital lymphedema in the remaining patient has remained stable during 10 years of followup. CONCLUSIONS: Congenital lymphedema of the genitalia is a challenging problem. Recurrences requiring multiple operations are common. We recommend expectant management of mild cases. In more severe cases excision without grafting should be attempted. While skin grafting may be the most definitive solution, it does not prevent recurrence in adjacent regions, and it carries the risk of skin contraction. Skin grafts should only be used when other techniques have failed. PMID- 9751401 TI - A boy with intrachromosomal triplication of the X chromosome. PMID- 9751402 TI - Refined microscopic urinalysis for red blood cell morphology in the evaluation of asymptomatic microscopic hematuria in a pediatric population. AB - PURPOSE: The use of refined microscopic urinalysis for the presence of dysmorphic red blood cells (RBCs) has been evaluated in children and adults with a known source of hematuria. We examined the clinical usefulness of this study in a pediatric population with an unknown source of hematuria. MATERIALS AND METHODS: Children 12 years old or younger referred for evaluation of asymptomatic microscopic hematuria exhibiting 4 or more RBCs per high power field were enrolled in this study. Patients provided a first morning urine sample subjected to refined urinalysis for RBC morphology. Standard evaluation of patients was performed until a final diagnosis of the hematuria source was identified. RESULTS: A total of 44 patients completed the study. Refined urinalysis revealed pure dysmorphic RBCs in 22 patients, pure isomorphic RBCs in 8 and mixed isomorphic/dysmorphic RBCs in 14. The presence of dysmorphic RBCs correctly predicted a glomerulotubular source of hematuria in 29 of 36 patients (sensitivity 83%, specificity 81%), while the presence of isomorphic RBCs predicted a uroepithelial source of hematuria in 2 of 8 patients (sensitivity 25%, specificity 22%). Hematuria and 2+ proteinuria (100 mg./dl.) were more sensitive (100%) and specific (83%) than the presence of dysmorphic RBCs in predicting glomerulotubular hematuria. CONCLUSIONS: We believe that this is a costly test offering little additional information to the evaluation of microscopic hematuria in children. A thoughtful history and physical examination with microscopic urinalysis and dipstick for proteinuria provide an equal amount of diagnostic information. We do not recommend its routine use in the evaluation of microscopic hematuria in children. PMID- 9751403 TI - Longitudinal cohort analysis of lethal prostate cancer progression in transgenic mice. AB - PURPOSE: Human prostate cancer is variably lethal, shows heterogeneous progression, and exhibits a spectrum of histopathology. Traditional rodent models of prostate cancer lack these characteristics. An alternative, autochthonous model of prostate cancer consists of transgenic mice which develop prostate cancer due to prostatic expression of SV40 T antigen. Lethal progression of such cancers in individual mice has not been previously characterized. Studies were undertaken to characterize the longitudinal progression of prostate cancers in these transgenic mice. METHODS: A prospective longitudinal cohort study was undertaken to characterize prostate cancer volume, progression, lethality, and histological heterogeneity in a transgenic mouse model of prostatic adenocarcinoma. Fifty-one transgenic mice were followed prospectively to determine the age at onset of palpable tumor and age at cancer-related death. Tumor volume was followed longitudinally by magnetic resonance imaging (MRI) in a subset of these mice and lethal cancers were evaluated by histopathology. RESULTS: Primary tumors became palpable at 10-38 weeks of age. Palpable tumors always preceded lethal progression. Cancer death followed 2-9 weeks later, and age at cancer death varied from 24 to 39 weeks of age. The histopathological changes were heterogeneous. Primary tumors were detectable by MRI before they became detectable by palpation. MRI showed that, analogous to human prostate cancers, volume of early stage primary tumors did not necessarily predict age at cancer death. CONCLUSION: Prostate cancer in transgenic mice mimics heterogeneic tumor progression in human prostate cancer, providing a uniquely relevant pre clinical model. Tumor detection by MRI and palpation are valid surrogate measures of tumor progression in this model. PMID- 9751404 TI - Osteopontin antisense oligonucleotide inhibits adhesion of calcium oxalate crystals in Madin-Darby canine kidney cell. AB - PURPOSE: We previously suggested that osteopontin (OPN) plays an important role in the process of deposited calcium crystals adhesion to cells in the early stages of urolithiasis. To further confirm this theory, we tried to inhibit OPN expression at the translational level and examined its cellular biological consequence on the formation and adhesion process of crystals. MATERIALS AND METHODS: We synthesized antisense and sense oligonucleotide corresponding to an appropriate part of the coding sequence for OPN in Madin Darby canine kidney (MDCK) cells. With the aid of lipofection reagent DOTAP, antisense and sense oligonucleotide were introduced into MDCK cells grown in a confluent monolayer. After further incubation, inhibition of OPN expression in the cells was assessed by immunofluorescence photomicrography, and formation of calcium oxalate crystals was quantitated by incorporation of 45Ca into the stone and visualized by scanning electron microscopy (SEM). RESULTS: Antisense oligonucleotide at concentrations higher than 20 microM inhibited synthesis of OPN. Incorporation of 45Ca into the calculus stone was inhibited by the addition of oligonucleotide in a concentration dependent manner in a range above 20 microM. More than 90% of incorporation was inhibited at 50 microM as compared to control. Inhibition of calcium crystal formation was confirmed by SEM. CONCLUSIONS: OPN was shown as a major component in the extracellular matrix involving the formation and adhesion of calcium crystals in the distal renal tubular cells, suggesting that OPN plays an important role in stimulating deposition and adhesion of calculus crystals to cells in the early stages of urolithiasis. PMID- 9751405 TI - Increase of low-affinity neurotrophin receptor p75 and growth-associated protein 43 immunoreactivities in the rat urinary bladder during experimentally induced nerve regeneration. AB - PURPOSE: Nerve regeneration in the urinary bladder after pelvic nerve plexus injury remains uncertain. The objectives were to establish a rat model of nerve regeneration in the bladder and to examine possible changes of low-affinity neurotrophin receptor p75 and growth-associated protein-43 (GAP-43) immunoreactivities during denervation and nerve regeneration. MATERIALS AND METHODS: Adult male rats were divided into 3 groups: controls, crush of the nerve bundles from the right major pelvic ganglion (MPG) to the bladder (nerve crush group) and removal of the right MPG (MPG removal group). Bladders were collected at 3, 7, 14, 30 and 60 days, and immunohistochemically stained for protein gene product 9.5 (PGP9.5: an axonal marker), p75 and GAP-43. RESULTS: In the nerve crush group, PGP9.5 positive nerves were decreased in number at 3 and 7 days, and then increased after 14 days in the muscle layer of the operated side. By 60 days, the density returned to control levels. However, MPG removal resulted in a decrease of the density of PGP9.5 positive nerves throughout the experimental periods. These findings indicate that nerve regeneration occurred in the nerve crush group. The density of p75 labeled fibers was significantly increased at 3 to 30 days postoperatively in the nerve crush group. p75 immunoreactivity showed smooth surface and cytoplasmic staining, indicating that Schwann cells were p75 positive. GAP-43 labeled fibers showed significantly greater density at 3 to 14 days postoperatively. Schwann cells were GAP-43 immunoreactive and, in particular, regenerating nerve fibers appeared to be GAP-43 positive at 14 days. CONCLUSION: The present study suggests that p75 and GAP-43 are involved in the mechanism(s) of nerve regeneration in the urinary bladder. PMID- 9751406 TI - Molecular analysis of collagens in bladder fibrosis. AB - PURPOSE: Fibrosis of bladder tissue is characterized by an abnormal deposition of connective tissue within different layers of the bladder wall, resulting in a low volume, high pressure vesical which may ultimately contribute to renal scarring and failure. These bladders are functionally referred to as "non-compliant" and may result from different etiologies: neurogenic, which encompasses myelodysplasia and spinal cord injury, or non-neurogenic, owing to obstruction or radiation therapy. To examine the molecular mechanisms responsible for this fibrosis, we have analyzed a well-characterized pediatric patient population for alteration(s) in collagen types I and III regulation at the protein and nucleic acid levels. MATERIALS AND METHODS: Immunohistochemical localization of collagen subtypes (I, III, and IV) was carried out using type specific monoclonal antibodies. Total collagen was determined by hydroxyproline analysis, and subtype specific expression of collagenous proteins, following cyanogen bromide extraction procedures, was quantified by competitive ELISA. Total RNA was extracted by guanidinium/phenol/chloroform, and slot blot hybridization analyses with radiolabeled human cDNA probes were quantified by densitometry of resulting autoradiograms. RESULTS: Connective tissue infiltration of detrusor smooth muscle bundles was specific for type III collagen. Protein analyses demonstrated: 1) an increase in total collagen, 2) a statistically significant increase in the type III: type I collagen ratio, and 3), an absolute increase in type III collagen protein in non-compliant bladder tissue. At the mRNA level, there was a coordinate increase in both collagen I and III steady-state mRNAs in non neurogenic bladder tissue, whereas neurogenic bladder tissue was characterized by an increase in the type III: type I mRNA transcript ratio. CONCLUSIONS: These data suggest that regulation of collagen synthesis in bladder fibrosis is complex and is characterized by both transcriptional and post-transcriptional mechanisms, depending upon the etiology of the fibrosis. PMID- 9751407 TI - Calcium oxalate crystal attachment to cultured kidney epithelial cell lines. AB - PURPOSE: Cultured kidney epithelial cell lines have frequently been used in urolithiasis research, and in particular in studies related to the interactions between stone crystals and cell membranes. There is evidence that when epithelial cell lines are transformed or serially passed to immortalize them, they experience changes in both cell physiology and morphology. Stone research utilizing cell cultures is frequently necessary due to the lack of an animal model for spontaneous stone disease. However, the interpretation of these cell culture research studies might be clouded by any significant differences in cell physiology between primary cells and continuous cell cultures. Therefore, the present study was conducted to compare calcium oxalate monohydrate (COM) crystal attachment to two primary kidney epithelial cell lines and to various continuous cell lines. MATERIALS AND METHODS: The cell lines surveyed were primary mouse proximal tubule cells (pMPT), primary inner medullary collecting duct cells (pIMCD), semi-continuous inner medullary collecting duct cells (cIMCD), BSC-1 cells, COS-1 cells, LLC-PK1 cells, MDCK cells, NRK-52E cells, and OK cells. All cell lines were cultured under identical conditions and the amount of COM attachment was measured using radioactive labeled COM crystals. RESULTS: COM crystal interaction with continuous kidney epithelial cells varied by a factor of two among the different cell lines. In general, cells that grew as regular, confluent cell monolayers, such as pMPT, pIMCD and cIMCD cells, exhibited the lowest levels of crystal attachment. Neither changes in membrane fluidity nor loss of normal epithelial cell membrane asymmetry seemed to correlate well with crystal attachment. After nine days of continuous cell culture, COM attachment to cIMCD cells dropped by 61 percent while crystal attachment to MDCK cells remained unchanged. It is unclear what makes these cell lines more resistant to crystal attachment compared to continuous cell lines. CONCLUSIONS: The significant difference in COM attachment between primary kidney epithelial cells and continuous epithelial cell cultures and the apparent differences in growth morphology between primary and continuous cell cultures must be considered when selecting a cell line for use in kidney stone research. Comparison of cIMCD cells and MDCK cells during extended culture time revealed one possible explanation for the differences in COM attachment: the formation of a mature, end-differentiated, non-dividing cell monolayer could protect the cells from crystal attachment. PMID- 9751408 TI - Telomerase activity, telomere length, and DNA ploidy in prostatic intraepithelial neoplasia (PIN). AB - PURPOSE: To investigate the relationship of telomerase activity, telomere length, and DNA ploidy in high grade prostatic intraepithelial neoplasia (PIN). MATERIALS AND METHODS: Tissue samples were carefully microdissected to obtain adenocarcinoma or PIN-containing tissue free of cancer. Telomerase activity was measured using the PCR-based telomeric repeat amplification protocol (TRAP). Telomere length was estimated from Southern blots of telomere restriction fragments (TRFs). DNA ploidy of PIN and carcinoma was determined by image analysis of adjacent Feulgen stained tissue sections. RESULTS: Telomerase activity was found in 4 of 25 samples (16%) of high grade PIN. All telomerase positive PIN foci had a diploid DNA content. Although 5 of 25 samples (25%) of high grade PIN foci analyzed were DNA aneuploid, none of these demonstrated telomerase activity. Telomerase positive foci of prostate carcinoma (69% of all cancer foci analyzed) displayed heterogeneity in TRF length, with a mean TRF length two kilobase pairs shorter than that of telomerase negative specimens. CONCLUSIONS: Telomerase activity is present in a low percentage of high-grade PIN foci, which are diploid by DNA content measurements. PMID- 9751410 TI - Time-course study of cellular immune response and testosterone metabolism in an autoimmune model for chronic prostatic inflammation. AB - PURPOSE: Little is known of the etiology and pathogenesis of chronic inflammatory prostate diseases of noninfectious origin. In our experimental autoimmune rat model for chronic prostatic inflammation (CPI) we evaluated, in a time-course study, the specific cellular immune response to male accessory glands (MAG) and metabolic activity in the prostate gland. Results obtained in CPI rats were compared with data from rats immunized with kidney homogenate as well as from non treated rats. MATERIALS AND METHODS: Specific cellular immune response against MAG antigen(s) was studied by delayed type hypersensitivity (DTH) and lymphocyte proliferation tests. The prostate 5alpha-reductase activity was studied in prostate homogenates by thin layer chromatography (TLC). RESULTS: DTH values were positive in MAG treated rats sacrificed at days 7 and 28 after first immunization (FI) (p < or = 0.05) in relation to kidney treated and non-treated rats. When we analyzed the proliferative responses to MAG antigen(s), an antigen specific proliferation, as shown by the mean [3H]thymidine uptake (cpm), was observed in rats sacrificed on days 14 and 28 (p < or = 0.05) after FI. The metabolic studies indicated that the 5alpha-reductase activity decreased slightly in MAG treated groups at day 14 after FI and diminished significantly at the end of CPI development. CONCLUSION: These data reveal that the prostatic endocrine cell destruction during CPI could be a consequence of immune/inflammatory cell mediated processes. PMID- 9751409 TI - Expression and tissue localization of membrane-types 1, 2, and 3 matrix metalloproteinases in human urothelial carcinomas. AB - PURPOSE: Three different membrane-type matrix metalloproteinases (MT1, 2, 3-MMP) which can activate proMMP-2 (progelatinase A) are thought to have an important role in various human carcinoma invasions and metastases. We examined the mRNA expression of MT-MMPs and the tissue immunolocalization of MT1-MMP in human urothelial carcinomas. MATERIALS AND METHODS: mRNA was extracted from 27 clinical urothelial carcinomas and 10 normal urothelial mucosa tissues remote from the tumor. RT-PCR using specific primers was performed, and PCR products were hybridized to 32P-labeled internal probes and analyzed by a bioimage analyzer. Immunolocalization was studied using a monoclonal antibody against MT1-MMP (114 6G6). RESULTS: MT1-MMP and MT2-MMP mRNA expressions in urothelial carcinomas were significantly higher than those in the normal mucosa. In contrast, MT3-MMP mRNA was little expressed in both tissues, and the amount of MT3-MMP mRNA appeared to be much lower than MT1-MMP and MT2-MMP in the tissue samples. In terms of the tumor multiplicity, MT1-MMP and MT2-MMP mRNA expressions in the group of multiple tumors were significantly higher than those in the solitary tumor group. The carcinoma cells were immunostained for MT1-MMP predominantly in invasive and superficial carcinoma cells. The immunoreactivity was more intense in the invasive type than in the superficial type. CONCLUSIONS: It is suggested that MT1 MMP and MT2-MMP play an important role in the development of human urothelial carcinomas and reflect some aspects of the pathogenesis of multifocal occurrence. In spite of the possible contribution to the invasive and metastatic phenotype, MT1-MMP mRNA and its product are thought to be expressed already in the clinical superficial stage in some cases of this tumor type. PMID- 9751411 TI - Neurogenically mediated cystitis in rats: an animal model. AB - PURPOSE: Pseudorabies virus (PRV) is a useful tool for mapping the control circuitry of the spinal cord. In the process of mapping CNS regulatory pathways for the lower urinary tract, a hemorrhagic change in the bladder was observed that was not overtly evident in other pelvic organs. The relationship between the appearance of hemorrhagic changes in the bladder and the evolution of PRV induced changes in the spinal cord was therefore explored. MATERIALS AND METHODS: Sprague Dawley rats were injected with PRV into the ACD tail-muscle. Bladder and CNS fixation were achieved by transcardial perfusion with formaldehyde. Multi-level sections were obtained from T8 through S4. Fixed tissue was stained and evaluated by light microscopy. Immunohistochemical stains were carried out for PRV and iNOS on spinal cord tissue. We were therefore able to evaluate the relationship between the manifestation of the hemorrhagic cystitis, appearance of the PRV in the spinal cord and evidence of CNS inflammation. RESULTS: The evolution of hemorrhagic cystitis paralleled the evidence of inflammation in the thoraco lumbar and sacral cord. These bladders contained 5 to 9 ml. of bloody urine (a normal rat bladder contains 1 to 2 ml.). On cystomanometry (CMG) the bladders were acontractile. No PRV could be cultured in the hemorrhagic bladders. The histological changes observed in the bladder represent true inflammation. CONCLUSION: There was no obvious explanation for these changes other than the associated inflammatory changes in the spinal cord. The findings are consistent with the hypothesis that a spinal cord stress, via an unknown metabolic pathway, can result in dramatic, neurogenically mediated changes in the bladder. PMID- 9751412 TI - Initiating genetic events in small renal neoplasms detected by comparative genomic hybridization. AB - PURPOSE: To identify the genetic alterations associated with renal adenomas. MATERIALS AND METHODS: We analyzed 37 renal adenomas obtained at autopsy (23 papillary and 14 non-papillary) by comparative genomic hybridization. RESULTS: In papillary tumors, the median number of gains and losses of genetic material per tumor was 2.0 and 1.0, respectively. Papillary tumors were characterized predominantly by gains of genetic material on chromosomes 7 (57%), 17 (35%), 16 (26%), 12 (26%), 3 (22%), 20 (22%) and loss of a sex chromosome (83%). In 6 papillary tumors less than or equal to 5 mm. in diameter, gain of chromosome 7 occurred in 4 specimens. Initiating events for papillary renal adenomas include gain of chromosome 7 and loss of a sex chromosome. In non-papillary tumors, the median number of gains and losses of genetic material per tumor was 1.0 and 1.0, respectively. Non-papillary tumors were characterized by loss of genetic material on chromosome 3p (50%), loss of a sex chromosome (36%) and a gain of chromosome 5 (43%). The initiating event for non-papillary renal adenomas is the loss of chromosome 3p. CONCLUSIONS: Renal adenomas demonstrate similar genetic alterations as clinically detected renal cell carcinomas. Their clinically indolent course may, in part, be a result of the lower number of genetic alterations per tumor than their clinically detected counterparts. Renal adenomas are thus small carcinomas which have not yet acquired the necessary genetic alterations leading to tumor progression. PMID- 9751413 TI - Interaction between CD44 and hyaluronic acid regulates human prostate cancer development. AB - PURPOSE: This study was designed to investigate the significance of the interaction between CD44 and hyaluronic acid in the development of human prostate cancer. MATERIALS AND METHODS: We transfected the standard CD44 isoform (CD44s) cDNA into PC3, a human prostate cancer cell line that barely expresses CD44s protein. The effects of the reintroduction of CD44s into PC3 cells on the ability to bind hyaluronic acid (HA) were analyzed by the cell adhesion assay and by the cell migration assay. The in vitro growth rate of CD44s transfected PC3 was measured by using the MTT assay. We then evaluated the in vivo tumor development of CD44s transfected PC3 cells by subcutaneous, intravenous, and intraperitoneal injection models in athymic nude mice. RESULTS: The introduction of CD44s in PC3 cells markedly enhanced the binding and migration of these cells to HA, but not to other extracellular matrix molecules. In vitro growth of CD44s-transfected PC3 was found to be significantly decreased. In addition, the CD44s-transfected PC3 cells also demonstrated reduced tumorigenicity and metastatic potential in in vivo experimental models. CONCLUSIONS: These findings suggest that the CD44s downregulation plays an important role in the development of human prostate cancer, in part through reduction of the ability to bind HA. PMID- 9751414 TI - Experimental partial unilateral ureter obstruction. I. Pressure flow relationship in a rat model with mild and severe acute ureter obstruction. AB - PURPOSE: To create an animal model with mild and severe partial unilateral ureter obstruction in young rats using a modified Ulm and Miller technique and to characterize the model by acute renal pelvic pressure measurements. MATERIAL AND METHODS: During anesthesia the upper fourth (n = 15) or the upper two-thirds of the left ureter (n = 15) was embedded into the psoas muscle causing either a mild or severe partial obstruction. Sham-operated control rats were prepared in parallel (n = 20). The baseline pelvic pressure, the perfusion pelvic pressure (perfusion rates: 0.2 to 1.0 ml. per minute) and peristaltic waves were recorded after a resting period. RESULTS: Mean baseline pelvic pressure and perfusion pelvic pressure were significantly higher in obstructed kidneys than in non obstructed kidneys, and significantly higher in severely obstructed kidneys than in mildly obstructed kidneys (p < 0.05). A fair linear relationship existed between the increase in pelvic pressure and the increase in perfusion rates in all groups. In the severely obstructed kidneys, baseline pelvic pressure was 16.9 +/- 2.3 cm. H2O and the perfusion pelvic pressure increased significantly from 41.7 +/- 3.3 cm. H2O to 68.6 +/- 6.3 cm. H2O. The pelvic peristaltic amplitude increased significantly following the increase in perfusion rate and there was a significant difference in the amplitude between severely obstructed and non obstructed kidneys when the perfusion rate exceeded 0.4 ml. per minute (p < 0.05). CONCLUSION: Embedding either the upper fourth or the upper two-thirds of the ureter into the psoas muscle produced a mild or a severe partial obstruction. In the latter model a significant increase in pelvic pressure and amplitude was observed, indicating the existence of severe obstruction. PMID- 9751415 TI - Restoration of sexual behavior and dopaminergic neurotransmission by long term exogenous testosterone replacement in aged male rats. AB - PURPOSE: We investigated the effects of long-term testosterone replacement on copulatory behavior and dopaminergic neurotransmission in the medial preoptic area of aged male rats. MATERIALS AND METHODS: The rats were divided into 3 groups depending on testosterone replacement. Those in the long-term replacement group were castrated at the age of 12 months and received testosterone replacement thereafter for 12 months. In the short-term replacement group, rats were castrated at the age of 22 months and high or low dose testosterone replacement was done for 2 months. The control group consisted of aged rats 24 months old and young rats 12 weeks old, neither of which had been castrated or received testosterone replacement. We observed sexual behavior in rats of these groups. After a behavioral test, we measured the tissue concentration of dopamine in the MPOA and the change rate of the extracellular dopamine level induced by infusion of N-methyl-D-aspartic acid (NMDA) in the MPOA and compared the long term replacement and no-replacement groups. RESULTS: The rats in the long-term replacement group showed a mount rate at the same level as that of young rats at 6 weeks after starting replacement and it was maintained to 24 months of age. Their mount rate was significantly higher than that of the rats with the short term replacement. A significantly higher change rate of dopamine release was recognized in the long-term group; however, no significant difference in the concentration of dopamine was recognized between aged rats with long-term replacement and those without replacement. CONCLUSIONS: Aged rats (24 months old) with long-term testosterone replacement maintained almost the same level of mount behavior as young rats (12 weeks old). The results imply that long-term testosterone replacement may favorably alter the decline in the process of sexual activity with aging. The restoration by testosterone replacement of dopaminergic activity in the MPOA may be involved in the maintenance of sexual function in aged rats. PMID- 9751416 TI - Dual-phase helical CT of the kidney: value of the corticomedullary and nephrographic phase for evaluation of renal lesions and preoperative staging of renal cell carcinoma. PMID- 9751417 TI - Radiosynthesis of [11C]Lu 29-024: a potential radiotracer for 5HT2 receptors PET studies. AB - For mapping 5-HT2 receptors in the central nervous system with positron emission tomography (PET), 2,5-dimethyl-3-(4-fluorophenyl)-1-(1-[11C]methyl-4-piperidinyl) 1H-indol e ([11C]Lu29-024) has been prepared. The precursor for the radiosynthesis of [11C]Lu29-024 was obtained in an overall yield of 53% by a convenient five-step synthesis; its reaction with [11C]methyl iodide afforded [11C]Lu29-024 in 35-50% radiochemical yield (decay corrected) in 45 to 50 min with a specific radioactivity ranging from 11 to 15 GBq/micromol. Following i.v. injections into rats, the analysis of plasma samples showed that the metabolism of [11C]Lu29-024 was rapid and extensive (60% of the original tracer was metabolized at 40 min). In contrast, only unmetabolized [11C]Lu29-024 could be detected in brain tissue. These biological results suggest that labeled metabolites have no access to brain tissue and further propose [11C]Lu29-024 as an interesting tool for PET studies of brain 5HT2 receptors. PMID- 9751419 TI - Synthesis and biodistribution of 64Cu-labeled monocationic diiminedioxime copper(II) complexes. AB - Monocationic copper(II) complexes of three derivatives of the diiminedioxime ligand 2,10-dioximino-3,9-dimethyl-4,8-diazaundeca-3,8-diene were labeled with 64Cu in high radiochemical yield, and the biodistribution of the complexes was measured in mice. The concentration of the complexes in the heart was low, but the partition coefficients of the complexes are less than optimal for a myocardial perfusion agent. All three complexes are resistant to reduction by glutathione in vitro. This suggests that more lipophilic derivatives of these compounds merit further investigation as possible myocardial perfusion agents. PMID- 9751418 TI - The in vivo behavior of copper-64-labeled azamacrocyclic complexes. AB - The use of copper radioisotopes in imaging and therapy applications has created a greater need for bifunctional chelates (BFCs) for complexing copper radioisotopes to biomolecules. It has been demonstrated that the charge and lipophilicity of the Cu-BFC complex has a significant effect on the in vivo behavior of the radiolabeled Cu-BFC-biomolecule conjugate. To evaluate the effects of charge, stability, and macrocyclic backbone size on the biological behavior of 64Cu complexes, a series of macrocyclic 64Cu complexes have been prepared, and the biodistributions of these agents were evaluated in normal Sprague-Dawley rats. Two macrocyclic backbones, dodecane and tetradecane, were evaluated; cyclen, DOTA, and DO2A were dodecane backbone derivatives, and cyclam, TETA, and et cyclam were tetradecane backbone derivatives. The biodistributions of the 64Cu labeled complexes correlated with differences in the size of the macrocycle backbone and the formal charge of the complex. All compounds showed uptake and clearance through the liver and kidneys; however, the positively charged 64Cu complexes showed significantly higher uptake in both of these organs than did the negatively charged or neutral complexes. 64Cu-TETA, a negatively charged complex with the tetradecane backbone, had the most efficient clearance by 24 hours' postinjection. These data suggest that negatively charged complexes may have more favorable clearance properties when used as BFCs. PMID- 9751420 TI - Is early sestamibi imaging in head and neck cancer affected by MDR status, p53 expression, or cell proliferation? AB - A consensus has emerged that early imaging (within 20 min post-injection [p.i.] of sestamibi) has the highest diagnostic yield. Tc-99m-sestamibi uptake has been associated with P-glycoprotein-mediated multi-drug resistance (MDR), tumor vascularization, invasiveness, and cell proliferation. The aim of this study was to assess whether these parameters affect tumor uptake in current imaging protocols. Twenty-three patients with squamous cell carcinoma (SCC) of the mouth floor who were scheduled for surgery were imaged 5 min. p.i. with a triple-head gamma camera (360 degrees, 3 degrees/step SPECT, UHRPAR collimators). Tumor:nontumor tissue (TBR), tumor:gingiva (TGR), tumor:salivary gland (TSR), and tumor:nuchal muscle ratios (TNR) were calculated based on the cts/pix values of ROIs in the axial slices. The expression of the MDR gene was determined histochemically from biopsies. Cell proliferation was quantitated by the histochemical analysis of the Ki-67 protein (Ki-67 index); additionally, the p53 tumor suppressor gene (p53 index), a posttranslational stabilizer of the cell cycle at the G1 stage, was assessed. No significant differences were found for the uptake indices between MDR+ and MDR- tumors. Moreover, no significant correlation between sestamibi uptake and the Ki-67 and p53 indices could be demonstrated. The diagnostic information content of currently applied imaging protocols for sestamibi in SCC of the mouth floor is not affected by tumor specific properties. PMID- 9751421 TI - Development of step-specific PET tracers for studying fatty acid beta-oxidation: biodistribution of [1-(11)C] octanoate analogs in rats and a cat. AB - To evaluate the potential of [1-(11)C]-3-(R,S)-methyloctanoate (BMOA), [1-(11)C] 2-octynoate, and [1-(11)C]-2-decynoate as PET tracers for studying particular steps in fatty acid beta-oxidation, we examined the pharmacokinetics of these compounds in rats and a cat. In rats given these compounds, high levels of radioactivity accumulated in the heart, liver, and kidneys, suggesting their potential as tracers for studying beta-oxidation in these tissues. These organs were clearly visible with PET in a cat given BMOA, indicating the utility of BMOA for imaging these organs. PMID- 9751422 TI - Tc(V)-DMS tumor localization mechanism: a pH-sensitive Tc(V)-DMS-enhanced target/nontarget ratio by glucose-mediated acidosis. AB - Since the conception of the pentavalent technetium polynuclear complex of dimercaptosuccinic acid, Tc(V)-DMS, a great number of papers published on its clinical applicability forced us to question "how tumor tissue appropriates the Tc(V)-DMS." Preliminary in vitro studies with Ehrlich ascites tumor cells (EATC) indicated the pH-sensitive character of this tumor agent. From this finding and the well-established notion that malignant tumors are more acidic than normal tissue, the in vivo correlation of Tc(V)-DMS accumulation in tumor tissue with its tissue acidification was considered of interest. The systemic lowering of tumor tissue pH by the stimulation of aerobic glycolysis has been well reported. In the present paper, the response of Tc(V)-DMS tumor accumulation to acidification induced by the glucose administration was explored in EATC-bearing mice. Measurement of tumor tissue pH was carried out by direct microelectrode technique and by histochemical umbelliferone technique in tumor tissue excised from EATC bearing mice. The regional acidity distribution is correlated with the regional radioactivity distribution registered by autoradiography. Evidence related to the pH sensitiveness of Tc(V)-DMS in response to glycolytic acidification was gathered; the pH measurement and the in vivo biodistribution of the double-tracer macroautoradiography with C-14 deoxyglucose (C-14-DG) demonstrated that the regional tissue distribution of Tc(V)-DMS was superimposed to that of C-14-DG. The glucose interventional modality offers the premier foundation for the interpretation of Tc(V)-DMS accumulation in diagnostic studies of malignant tumors. PMID- 9751423 TI - Increased streptavidin uptake in tumors pretargeted with biotinylated antibody using a conjugate of streptavidin-fab fragment. AB - Radiolabeled streptavidin accumulated in tumors pretargeted with biotinylated antibody. However, the absolute delivery of radioactivity was limited. To increase the tumor uptake of radioactivity further, we conjugated streptavidin with a mouse monoclonal antibody (MAb) fragment, OST6Fab, which recognizes antigen on human osteosarcoma. Another mouse MAb, OST7, which also reacts with the same tumor but recognizes an epitope different from the OST6 epitope, was biotinylated. The radioiodinated streptavidin-OST6Fab conjugate was administered to tumor-bearing mice after the biotinylated OST7 pretargeting. The uptake of the conjugate in tumors pretargeted with the biotinylated antibody was significantly higher than that of streptavidin and that of the conjugate of streptavidin and irrelevant Fab fragment. Renal uptake of radioactivity was decreased markedly, and the blood clearance was retarded by the conjugation with Fab fragment. In conclusion, the conjugate of streptavidin with specific Fab fragment increased the accumulation of radioactivity in tumors pretargeted with biotinylated antibody. PMID- 9751424 TI - Use of 99mTc-mercaptoacetyltriglycine (MAG3)-biocytin hepatobiliary scintigraphy to study the protective effect of a synthetic enzyme inhibitor on acute hepatotoxicity in mice. AB - Recent data suggest that inhibitors of ethanol-inducible cytochrome P450 (CYP2E1) can protect the liver from injury caused by various substrates of CYP2E1. In this study, we measured the protective effect of isopropyl-2-(1,3-dithioetane-2 ylidene)-2[N-(4-methylthiazol -2-yl)-carbamoyl]acetate (YH439), a transcriptional inhibitor of CYP2E1, against carbon tetrachloride (CCl4)-induced hepatotoxicity by using various conventional methods and dynamic scintigraphy with 99mTc mercaptoacetyltriglycine (MAG3)-biocytin, a recently developed scintigraphic agent. Balb/c mice were pretreated with two doses of YH439 (50 or 150 mg/kg per day) at 48 h and 24 h and one dose of CCl4 (0.25 mL/kg) at 18 h before scintigraphy. The results were compared with those of two other groups, one that received CCl4 but not YH439, and the other that received neither (control). Scintigraphic images were acquired continuously at 15-sec intervals for 30 min. Pharmacokinetic parameters, such as peak liver/heart ratio (r(max)), peak liver uptake time (t(max)), and hepatic half-clearance time (HCT), were obtained from time-activity curves derived from regions-of-interest (ROI) over the liver and the heart. Acute administration of CCl4 alone caused centrilobular necrosis and serum transaminase levels to rise more than 5 times higher than those of the control group. Pharmacokinetic parameters also changed significantly from those of the control group. Administration of YH439 prevented centrilobular necrosis and significantly improved pharmacokinetic parameters. This study demonstrates for the first time that hepatobiliary scintigraphy can be used to study in vivo biochemistry of the CYP2E1 inhibitor (YH439) against liver toxicity. PMID- 9751425 TI - Importance of the two ester functions for the brain retention of 99mTc-labelled ethylene dicysteine diethyl ester (99mTc-ECD). AB - 99mTc-ethylene dicysteine diethyl ester (99mTc-L,L-ECD) is a neutral lipophilic tracer agent that crosses the blood-brain barrier and is retained in the brain of primates following enzymatic hydrolysis of one of the ester functions to the ionized mono-ester, mono-acid metabolite. Up to now, it is not clear whether the second ethylcarboxylate group is essential for brain uptake and retention. Therefore, we have synthesized and studied two derivatives of 99mTc-L,L-ECD that contain only one ethylcarboxylate function, namely 99mTc-labelled L- and D ethylene cysteamine cysteine ethyl ester (99mTc-ECCE). Direct labelling of L- or D-ECCE at neutral pH and room temperature resulted for each of them in the formation of two probably diastereomeric 99mTc-complexes in a 1:1 ratio. This means that four different isomers could be isolated. The 99mTc-labelled complexes formed after labelling ECCE (A and B, in order of elution during HPLC) are slightly less lipophilic than 99mTc-L,L-ECD. In mice, all four isomers show a low brain activity at 10 min post injection (p.i.), approximately 0.1% to 0.3% of the injected dose versus 0.9% for 99mTc-L,L-ECD. The clearance from the blood is comparable with (isomers LA and LB) or slower (isomers DA and DB) than that of 99mTc-L,L-ECD. Isomers LA and DA show high liver uptake and rapid excretion to the intestines. Both 99mTc-ECCE-LB and 99mTc-ECCE-DB, the most lipophilic isomers, are cleared from the blood mainly by the kidneys and excreted more efficiently to the urine. 99mTc-ECCE-LB is characterized by a surprisingly high heart uptake (about 1.5% of i.d.) at 10 min p.i. versus 1.0% for 99mTc methoxyisobutylisonitrile (99mTc-MIBI) and 0.2-0.3% for the other isomers, but also lung uptake is relatively high. In the baboon, brain and heart uptake of isomers LA and LB of 99mTc-ECCE were negligible. Activity concentrated mostly in the hepatobiliary system for isomer LA and in the renal system for isomer LB. The results indicate a clear difference in biological behaviour between 99mTc-L,L-ECD and the four isomers of 99mTc-ECCE. This shows that the presence of both ester functions in 99mTc-L,L-ECD is an essential structural requirement for its brain uptake and retention in primates. PMID- 9751426 TI - 198Au-labeled hydroxymethyl phosphines as models for potential therapeutic pharmaceuticals. AB - The development of novel gold-198 complexes with water-soluble phosphines is reported. A series of cationic and hydrophilic 198Au complexes containing the ligands tris(hydroxymethyl)phosphine (THP, 1) 1,2 bis[bis(hydroxymethyl)phosphino]benzene (HMPB, 2), and 1,2 bis[bis(hydroxymethyl)phosphino]ethane (HMPE, 3) were prepared and evaluated as models for potential gold-199 radiopharmaceuticals. The 198Au complexes were formed in high radiochemical purity by simply mixing H198AuCl4 with the respective ligand. The complexes were shown to exhibit high in vitro stability over wide pH ranges and temperatures. However, only the 198Au(HMPB)2+ complex was found to exhibit good in vivo stability. HPLC analyses indicated that the 198Au complexes with these three phosphine ligands produced singular species with similar retention times as compared to their known macroscopic complexes. PMID- 9751427 TI - A kit formulation for preparation of [(111)In]In-DTPA-folate, a folate-receptor targeted radiopharmaceutical. AB - A kit formulation has been developed for convenient, routine compounding of (111)In-labeled In(III)-DTPA-Folate, an investigational radiopharmaceutical for targeting tumor-associated folate receptors. The kit consists of the DTPA-Folate conjugate in sodium citrate solution, from which [(111)In]In-DTPA-Folate can be rapidly and reliably compounded by the addition of aqueous [(111)In]In(III) chloride. PMID- 9751428 TI - Proliferating cell nuclear antigen expression is increased in small bowel epithelium in the elderly. AB - Although previous studies suggest that in aging animals the small intestine is in a hyperproliferative state, no information is currently available on the influence of age on the proliferation pattern of human small bowel enterocytes. The immunohistochemical expression of the proliferating cell nuclear antigen (PCNA), the villous height to total mucosal thickness ratio and the enterocyte height were evaluated in a panel of duodenal biopsy specimens obtained from 18 subjects aged less and 14 subjects aged more than 65 years. There was a significant positive correlation (P < 0.001) between age in years and percent of positive PCNA enterocytes both at the level of crypts (rs = 0.50) and villi (rs = 0.77). Moreover, the percentage of PCNA+ enterocytes was significantly higher in elderly versus adult subjects, both at the level of villi (6.5 vs 0%; P < 0.001) and of crypts (40.0 vs 23.7%; P < 0.01). No correlation was found between the percentage of PCNA + enterocytes and enterocyte height or villous height to total mucosal thickness ratio. Our results show that PCNA reactivity increases with advancing age both in crypts and villi. This abnormality of the proliferation pattern may explain the coexistence of normal morphology and impaired absorptive function in the elderly. PMID- 9751429 TI - Efficacy of anti-influenza peptide vaccine in aged mice. AB - Influenza infections may cause serious morbidity, as well as mortality in the elderly. In the present study we vaccinated old and young mice of two strains with three synthetic recombinant constructs (Levi and Arnon, 1995. In: Chanock, R.M. et al. (Eds.), Vaccines 95. CSHL Press, New York, pp. 311-316) and examined their capacity to eliminate a challenge of virus. Virus clearance from the lungs in the aged was very efficient, although the immune response in the aged was comparatively reduced. The data demonstrate that an intranasal administration of peptide-based anti-influenza vaccine without any adjuvant can be efficient and protective in old mice. Further studies are needed to determine whether such constructs will provide an effective vaccine for elderly human subjects. PMID- 9751430 TI - Nutritional influences on immune response in healthy aged persons. AB - Healthy elderly (80+/-5 years) with different nutritional status were compared to young healthy adults (25+/-5 years) to quantify the relative influences of aging and nutrition on immune response. Aged persons, without alteration of their nutritional status, had lower CD3+, CD8+, and CD45RA+ as well as higher CD2+CD3-, CD2+CD4-CD8-, and CD45RO+ T cell subsets and IL-6 release than their younger counterparts. T cell proliferation and IL-2 production were comparable in the two healthiest groups. Aged subjects with low nutritional status expressed similar but more marked changes in immune response while nutritional status did not influence the immune response in young subjects. Furthermore, lower nutritional status was associated with lower CD4+ counts and lower T cell functions in aged persons. These results indicate that the influences of aging and undernutrition in humans are cumulative and suggest that some changes in immune response that have been attributed to aging may, in fact, be related to nutrition and not aging. PMID- 9751431 TI - Effect of age on mitogen induced protein tyrosine phosphorylation in human T cell and its subsets: down-regulation of tyrosine phosphorylation of ZAP-70. AB - Several events of T cell activation have been reported to decline in humans with age. Since protein tyrosine phosphorylation is an early critical event of T cell activation, we performed a systematic analysis of the age-associated changes in the mitogen induced protein tyrosine phosphorylation of human T lymphocytes using SDS-PAGE and Western blotting techniques. Following stimulation with Con A and PHA, an identical pattern of protein tyrosine phosphorylation was observed in the lysates of T cells prepared from seven healthy young adults and eight healthy elderly human subjects. Five different high molecular mass proteins (75, 115, 120, 140 and 170 kDa) were consistently tyrosine phosphorylated in all of the donors from both age groups and peaked between 3 and 10 min. Tyrosine phosphorylation of the above substrates was observed in both CD4 and CD8 subsets. When compared for individual donors from both age groups, variations in the T cell response with regard to net tyrosine phosphorylation for all the substrates was observed. However, the mitogen induced level of tyrosine phosphorylation of only p75 was found to be significantly lower in unfractionated T cells as well as CD4 and CD8 subsets of older subjects than that of young subjects. Using immunoblotting, p75 was identified as ZAP-70, a member of the syk family of protein tyrosine kinases. Understanding of the biochemical basis of the reduced level of tyrosine phosphorylation of ZAP-70 will be helpful in delineating the molecular basis of age-associated impairment of T cell activation. PMID- 9751432 TI - Effect of age and dietary restriction on expression of heat shock protein 70 in rat alveolar macrophages. AB - Dietary restriction (DR) is the only effective experimental manipulation known to retard aging in rodents, and this manipulation has been shown to alter a variety of processes that change with age. However, there is no information on the effect of DR on macrophage Function. In the present study, the effect of aging and DR on the ability of alveolar macrophages (AMs) to express the heat shock gene, hsp70 was studied. AMs were isolated by lavage from the lungs of young (4-6 months) and old (24-26 months) rats fed either ad libitum (AL) or a restricted diet (60% of AL). There was no age-related change in the number of cells recovered from young and old rats fed AL. However, the number of cells recovered from the lungs of the DR rats was reduced, and this decrease was statistically significant in young rats. The expression of heat shock protein 70 (hsp70) was measured by the level of the hsp70 mRNA transcript in total RNA isolated from AMs cultured under two conditions: in suspension and after adherence to plastic. When AMs were incubated at 37 degrees C in suspension, no detectable hsp70 expression was observed; however, hsp70 expression was induced at 37 degrees C when the AMs adhered to the plastic culture dishes. Hsp70 mRNA levels were rapidly induced by heat shock (43 degrees C, 1 h) in AMs cultured both in suspension and on plastic. The induction of hsp70 expression did not change significantly with either age or DR in AMs cultured in suspension. In contrast, the induction of hsp70 mRNA levels by AMs adherent to plastic culture plates decreased approximately 70% with age, and hsp70 induction was greater in AMs isolated from DR rats; this difference was statistically significant in young rats. The induction of hsp70 by heat shock (43 degrees C, 1 h) also decreased with age in the adherent AMs, and DR increased the induction of hsp70 expression three- to fourfold in adherent AMs from both young and old rats. PMID- 9751433 TI - Developmental enhancement of secretory response to isoproterenol coupled with increases in beta-adrenoceptor density and Gs protein function in rat parotid tissues. AB - The beta-adrenergic agonist, isoproterenol (IPR), stimulated more significantly and sensitively amylase secretion from both the tissues of 7- and 56-day-old rats than a cholinergic agonist, carbachol, at the same concentration. The EC50 value of amylase secretion with IPR decreased significantly during development but that with carbachol did not change. Estimation by measuring bindings of [3H]dihydroalprenolol and [3H]quinuclidynylbenzylate indicated the marked increases in the numbers of both beta-adrenoceptors and muscarinic receptors in the tissues during development. The affinity of beta-adrenoceptors for the agonist was also enhanced during development, but that of muscarinic receptors for the agonist was not. These developmental changes in the number and affinity of beta-adrenoceptors and muscarinic receptors paralleled those in amylase secretory response of the tissues to their agonists. The response of adenylate cyclase (AC) of the tissues to 1 microM IPR was steadily enhanced after birth. In contrast, the response of AC to 1 microM forskolin was high until 14 days old, but markedly decreased at 28 days old and thereafter maintained this level. The increase in cholera toxin-catalyzed ADP-ribosylation (AR) of stimulatory GTP binding proteins (Gs proteins) in the tissues was apparent at 14 days old, reaching a maximum at 56 days old and thereafter decreasing with age. On the other hand, pertussis toxin-catalyzed AR of inhibitory GTP binding proteins (Gi proteins) did not change after birth. Thus, the ratio of apparent levels of Gs to Gi proteins increased significantly after birth, reaching a maximum at 56 days old, but decreased rapidly till 84 days old and thereafter maintained this level. These changes in the ratio paralleled those in the response of AC to IPR. These results showed that the rapid and marked increases in the number and affinity of beta-adrenoceptors and the ratio of apparent levels of Gs to Gi proteins in rat parotid tissues during development had a key role in the enhancement of the secretory response of the tissues to beta-agonists. PMID- 9751434 TI - Age and exercise-related changes in lipid peroxidation and superoxide dismutase activity in liver and soleus muscle tissues of rats. AB - Thiobarbituric acid-reactive substances (TBARS) level, as marker of lipid peroxidation, and superoxide dismutase (SOD) activity, as endogenous antioxidant enzyme, were examined in liver and soleus muscle tissue of young and old male Wistar rats. We established different types of exercise running on a treadmill both for young and old rats, investigated the effect of aging, exhaustion and training on these groups. The hepatic TBARS levels were raised in the short training young group and in the long-training old group. On the other hand, the TBARS content decreased in soleus muscle in the short-training young group, and long-training exercise enhanced lipid peroxidation in old rats. SOD activity increased in liver in short-training group. while this activity showed the lowest values in long-training old rats. With respect to soleus muscle tissue, SOD activity was elevated after exhaustive exercise in young rats and old rats had the highest activity in the long-training old group. These findings suggest that free radicals play a role in aging and that the different type, intensity and duration of exercise modify the lipid peroxidation level and antioxidant enzyme activity. PMID- 9751435 TI - Host defenses in the aged: evaluation of the balance between oxidizing species generation and reducing power in phagocyting human granulocytes. AB - The respiratory burst reaction has been studied in human granulocytes from normal subjects divided in four age groups: (I) 20-29, (II) 30-39, (III) 40-49, and (IV) 50-59 years old. Zimosan opsonized particles (OZ) were used to evaluate, simultaneously, the reducing power and the oxidizing species generation. Our results showed a strong parallelism and a direct correlation between oxidizing species generation and reducing power in the groups (I) and (II), in presence or in the absence of opsonized zimosan. However, the age groups (III) and (IV) showed an increase in the free radicals generation and a significant decrease in the cellular reducing power. This inverse correlation observed between oxidizing/reducing power in the (III) and (IV) age groups may suggest a metabolic cellular disequilibrium. PMID- 9751436 TI - Recognition of prosody following unilateral brain lesion: influence of functional and structural attributes of prosodic contours. AB - The perception of prosodic distinctions by adults with unilateral right- (RHD) and left-hemisphere (LHD) damage and subjects without brain injury was assessed through six tasks that varied both functional (i.e. linguistic/emotional) and structural (i.e. acoustic) attributes of a common set of base stimuli. Three tasks explored the subjects' ability to perceive local prosodic markers associated with emphatic stress (Focus Perception condition) and three tasks examined the comprehension of emotional-prosodic meanings by the same listeners (Emotion Perception condition). Within each condition, an initial task measured the subjects' ability to recognize each "type" of prosody when all potential acoustic features (but no semantic features) signalled the target response (Baseline). Two additional tasks investigated the extent to which each group's performance on the Baseline task was influenced by duration (D-Neutral) or fundamental frequency (F-Neutral) parameters of the stimuli within each condition. Results revealed that both RHD and LHD patients were impaired, relative to healthy control subjects, in interpreting the emotional meaning of prosodic contours, but that only LHD patients displayed subnormal capacity to perceive linguistic (emphatic) specifications via prosodic cues. The performance of the RHD and LHD patients was also selectively disturbed when certain acoustic properties of the stimuli were manipulated, suggesting that both functional and structural attributes of prosodic patterns may be determinants of prosody lateralization. PMID- 9751437 TI - The influence of semantic and perceptual encoding on recognition memory in Alzheimer's disease. AB - Previous results from a population of patients with Alzheimer's disease (Dalla Barba and Goldblum, 1996) demonstrated that the ability of patients to make a semantic association between two items was significantly and positively correlated to their performance on a yes/no recognition task for the same items and that patients who were impaired on the semantic task did significantly worse on the recognition task than patients who were unimpaired on the semantic task. These findings gave support to a hierarchical model of organization of human memory in which episodic memory depends on the integrity of semantic memory. The present study further investigates the relationship between semantic memory deficits and episodic recognition memory in 15 patients with Alzheimer's disease and 15 controls, as a function of their semantic and perceptual encoding abilities and of their cognitive impairment in other domains. The results confirmed the previous findings and showed that, although patients heavily relied on perceptual analysis, this type of encoding did not enhance their recognition memory. Correlations analyses showed that some patients who were not impaired in the semantic association, but with particularly low scores on a verbal fluency task presented with a pattern, in recognition memory tasks, that suggests a possible early involvement of frontal lobes in this subgroup of patients. PMID- 9751438 TI - Stimulus-response compatibility in representational space. AB - Spatial stimulus response (S-R) compatibility designates the observation that speeded reactions to unilateral stimuli are faster for the hand ipsilateral than for the hand contralateral to the sensory hemifield containing the stimulus. In two experiments involving presentation of the numbers 1 to 11 in the center of the visual field we show (1) a left-hand reaction time (RT) advantage for numerals < 6 and a right-hand advantage for those > 6 for subjects who conceive of the numbers as distances on a ruler, and (2) a reversal of this RT advantage for subjects who conceive of them as hours on a clock face. While the results in the first task (RULER) replicate a robust finding from the neuropsychology of number processing (the "SNARC effect") those in the second task (CLOCK) show that extension of the number scale from left to right in representational space cannot be the decisive factor for the observed interaction between hand and number size. Taken together, the findings in the two tasks are best accounted for in terms of an interaction between lateralized mental representations and lateralized motor outputs (i.e. an analog of traditional spatial S-R compatibility effects in representational space). We discuss potential clinical applications of the two tasks in patients with neglect of representational space. PMID- 9751439 TI - Understanding ambiguous words in sentence contexts: electrophysiological evidence for delayed contextual selection in Broca's aphasia. AB - This study investigates whether spoken sentence comprehension deficits in Broca's aphasics results from their inability to access the subordinate meaning of ambiguous words (e.g. bank), or alternatively, from a delay in their selection of the contextually appropriate meaning. Twelve Broca's aphasics and twelve elderly controls were presented with lexical ambiguities in three context conditions, each followed by the same target words. In the concordant condition, the sentence context biased the meaning of the sentence-final ambiguous word that was related to the target. In the discordant condition, the sentence context biased the meaning of the sentence-final ambiguous word that was incompatible with the target. In the unrelated condition, the sentence-final word was unambiguous and unrelated to the target. The task of the subjects was to listen attentively to the stimuli. The activational status of the ambiguous sentence-final words was inferred from the amplitude of the N400 to the targets at two inter-stimulus intervals (ISIs) (100 ms and 1250 ms). At the short ISI, the Broca's aphasics showed clear evidence of activation of the subordinate meaning. In contrast to elderly controls, however, the Broca's aphasics were not successful at selecting the appropriate meaning of the ambiguity in the short ISI version of the experiment. But at the long ISI, in accordance with the performance of the elderly controls, the patients were able to successfully complete the contextual selection process. These results indicate that Broca's aphasics are delayed in the process of contextual selection. It is argued that this finding of delayed selection is compatible with the idea that comprehension deficits in Broca's aphasia result from a delay in the process of integrating lexical information. PMID- 9751440 TI - The role of the corpus callosum in visual orienting: importance of interhemispheric visual transfer. AB - In a tachistoscopic visual search task, the effects of ipsi- and contralateral distractors on target search were investigated in two complete commissurotomy patients. Pop-out distractors slowed the search for contralateral targets in both patients, i.e. search was not independent in both hemifields. In normals, we previously observed an extinction-like asymmetry in that distractors in the right visual hemifield interfered with target search in the left visual hemifield, but not vice versa. This pattern was also found in one of our patients, N.G., whereas the other, L.B., showed a reversed laterality effect. While N.G. is able to transfer visual shape information between hemispheres, L.B. is not. The data suggest that the reversal of the contralateral distractor asymmetry in L.B. is due to the disruption of ipsilateral visual input to the right hemisphere. PMID- 9751441 TI - Naming in semantic dementia--what matters? AB - One of the major symptoms of semantic dementia (or progressive fluent aphasia) is profound word-finding difficulties. We present here a cross-sectional study of the factors affecting picture naming in semantic dementia based on data obtained from eight patients, together with a longitudinal analysis of naming in another patient. Various properties and attributes of the objects were entered into a series of regression analyses in order to predict which items the patients could or could not name. The analyses showed that object familiarity, word frequency and age-of-acquisition predicted naming success for the group and, in most cases, for each individual patient, irrespective of lesion site or overall naming success. We propose that the pattern of naming in semantic dementia is best described in terms of reduced semantic activation within a cascading/interactive speech production system. We suggest that object familiarity, and possibly word frequency, reflect the inherent robustness of individual semantic representations to the decay process in terms of both quantity and quality of experience. Age-of acquisition and word frequency (at a phonological-lexical level) predicts naming success, because frequent, early-acquired words are relatively easy to activate even with reduced semantic "input". PMID- 9751442 TI - Recall and recognition memory in patients with focal frontal, temporal lobe and diencephalic lesions. AB - Patients with frontal, temporal lobe, or diencephalic lesions were compared with healthy controls on measures of recall and recognition memory for word lists. Exposure times were titrated to match recognition memory scores 30 s after the end of word-list presentation as closely as possible. Using this technique, we failed to find a disproportionate impairment in recall memory in either the frontal lobe lesion patients or in the amnesic (temporal lobe and diencephalic) patients, compared with healthy controls. Consistent with this finding, performance on these tasks showed highly significant correlations with anterograde memory quotients (despite the titration procedure), but not with executive/frontal function tasks. On the other hand, the frontal lobe lesion group showed disproportionate benefit in the recall of semantically categorised words, compared with unrelated words. This may indicate an impairment in retrieval or access, compared with the amnesic (temporal lobe and diencephalic) patients, and/or an inability to organise their learning of unrelated words spontaneously, compared with healthy controls. PMID- 9751443 TI - The judgement of absence in neglect. AB - The usual way of looking at neglect is by investigating how neglect patients fail to detect that something is there. In this study, we look at how neglect patients correctly detect that something is not there. Patients with parietal lesions (11 with and 16 without neglect) and 23 control subjects indicated whether a dot target was or was not present in a geometrical display. While control subjects were consistently (and unexpectedly) faster in the no-dot than in the dot condition, the distinguishing response time pattern of right parietal patients with neglect was not--as one might expect--a relatively longer response time to left vs right targets, but a longer response time to target absence vs presence. This may be due to a serial search or, alternatively, it might result from double checking for target absence, produced by lowered perceptual confidence. Since this "wariness" about stimulus absence seems to operate in parallel with neglect patients' denial of the deficit, we conclude that the response time pattern observed in this study could be used as a measure of subjective (un)awareness of neglect. PMID- 9751444 TI - A reversal of the temporal gradient for famous person knowledge in semantic dementia: implications for the neural organisation of long-term memory. AB - On tests of autobiographical memory, patients with semantic dementia demonstrate significantly better retrieval of episodic events from the recent past compared with the distant past. This reversal of the Ribot effect has been attributed to the relative sparing of the hippocampal complex in the disorder. Current computational models of long-term memory predict a similar time-dependent pattern of impairment on tests of remote semantic memory. Five patients with semantic dementia were tested on recognition (familiarity) and identification (knowledge) of famous names selected from four different time-periods: 1950's, 1980's, 1990 1993 (early 1990's) and 19941996 (current). As expected, it was found that one patient DM (who had focal left temporal lobe atrophy) showed no significant impairment on recognition of famous names, but was significantly better at producing information about people who were currently famous compared to people famous in the other three time-periods. The other four patients (who had bilateral temporal lobe damage) showed better recognition of famous names from the current time-period (and to a lesser extent the 1950's), yet were profoundly impaired on the identification component, producing very little information across all four time-periods. The results are discussed with respect to current views of the neural organisation of person-specific and general semantic memory. PMID- 9751445 TI - Stereochemical studies of the isomerization of novel 2-alkyl-9-phenyl-2,3,4,4a- and 2,3,4,9-tetrahydro-1H-indeno[2,1-c]pyridines. AB - The published synthetic route to the antihistaminic tetrahydroindeno[2,1 c]pyridines (phenindamines) relies on catalytic reduction of the precursor dihydroindenopyridines. This reduction gives mixtures of 9,9a- and 4a,9a-enes and the clinically active 4a,9a isomer has to be isolated by recrystallization of an appropriate salt. The structure of the product recovered depends on the anion used to isolate the proton salt and appears to be arbitrary. To rationalize this outcome a series of novel N-2 alkylated tetrahydroindeno[2,1-c]pyridines and their diene precursors has been synthesized from accessible piperidines. The structures and geometry of the piperidines and the dihydro- and tetrahydroindenopyridines have been determined by 1H and 13C NMR. An unusual feature of the proton spectra of the piperidines is the resonance of the axial protons at lower field than their equatorial counterparts. By controlling the reaction conditions for the reduction of the dihydroindenopyridines to their tetrahydro derivatives the kinetic or thermodynamic product can be selected as required. A predictable outcome for the reductions investigated was achieved and is generally applicable. PMID- 9751446 TI - Solution-structure of a peptide designed to mimic the C-terminal hexapeptide of endothelin. AB - Ac-cyclo(Cys-His-Leu-Asp-Cys)-Ile-Trp-OH, has been designed by computer-aided molecular-modelling techniques to mimic the proposed alpha-helical conformation of the C-terminal hexapeptide of endothelin. Two-dimensional proton nuclear magnetic resonance spectra were acquired for the peptide dissolved in d6-DMSO or D2O-H2O and the distance and angle constraints incorporated into simulated annealing experiments. Conformers generated from the D2O-H2O data superposed on the corresponding main-chain atoms in the crystal structure of endothelin 1 and the solution structure of BQ-123 with root mean square co-ordinate differences of 0.9A and 0.77A, respectively. The peptide did not elicit antagonism of endothelin induced in-vitro contractions of rabbit aorta (endothelin A receptor) or rabbit bronchus (endothelin B receptor) preparations. Because the peptide can adopt a conformer which closely matches the equivalent residues in the endothelin 1 crystal structure and in BQ-123, we suggest BQ-123 does not necessarily mimic the endothelin C-terminal region to achieve its antagonism, and that a helical conformation of the endothelin C-terminal hexapeptide does not favour its interaction at the endothelin B receptor. PMID- 9751447 TI - Optimization of sustained-release diltiazem formulations in man by use of an in vitro/in-vivo correlation. AB - In-vitro/in-vivo correlations (IVIVC) are useful for predicting in-vivo results from in-vitro data. An IVIVC has been used to optimize a hydrocolloidal-based matrix tablet designed to be bioequivalent to an existing once-daily diltiazem HC1 product (Dilacor XR 240mg; Rhone-Poulenc Rorer). Data from a preliminary formulation dosed to fasted and fed subjects were used to establish the IVIVC. The correlation was then used during reformulation of the dosage forms to predict changes in the maximum plasma concentration (Cmax) and the area under the plasma concentration-time curve (AUC) for fasted and fed subjects using in-vitro dissolution data. The IVIVC adequately predicted plasma profiles of two optimized formulations in studies with fasted and fed subjects. PMID- 9751448 TI - Prediction of serum vancomycin concentrations using one-, two- and three compartment models with implemented population pharmacokinetic parameters and with the Bayesian method. AB - Although previous studies have shown that vancomycin has a complicated pharmacokinetic profile requiring description using a two- or, better, three compartment model, until recently predictions of serum vancomycin concentrations have been mainly based on one- or two-compartment models using computer software packages. In this study, we have predicted serum vancomycin concentrations in 59 patients using one-, two- and three-compartment models with implemented population pharmacokinetic parameters in the Abbott PKS program and by use of the Bayesian method. The percentage errors of predictions made using the one compartment model were smaller when either the Bayesian method or implemented population pharmacokinetic parameters were used (medians of -8.61% and -9.49%, respectively). Predictions using the one-compartment model with the Bayesian method were less biased (median of -1.52 microgmL(-1). The best predictions were those made using the three-compartment model with the Bayesian method-they were most accurate (median of 3.40 microgmL(-1) and highly precise (median of 11.53 microg(2)mL(-1)). The results suggest that predictions made using the one compartment model with implemented population pharmacokinetic parameters are preferable if no samples are available, otherwise predictions made using the three-compartment model with the Bayesian method are preferable. The results also supported our previous argument that the greater the number of compartments involved in individualization, the better the predictions obtained using the Bayesian method. PMID- 9751449 TI - Enterohepatic cycling and pharmacokinetics of oestradiol in postmenopausal women. AB - The pharmacokinetics and enterohepatic cycling of oestradiol have been studied after three oral, single-dose administrations of equimolar doses of oestradiol alone, oestradiol plus desogestrel and oestradiol valerate, in a 3-way cross-over mode in 18 healthy postmenopausal women. Oestradiol was readily absorbed and metabolized to oestrone, which reached much higher serum concentrations (140pgmL( 1)) than its parent compound (35pgmL(-1)). All three formulations had the same kinetic profile and were bioequivalent on testing. Noticeable first and second absorption phases were apparent from the oestradiol and oestrone serum concentration-time curves for all oestradiol formulations. The mean serum concentration-time curves of the metabolite oestrone (corrected for endogenous oestrone) showed a second maximum at approximately 25h. By means of line feathering, serum concentration-time curves were constructed which belonged to the first, second and third phases of absorption. The maximum serum concentration, Cmax, of the second absorption or recirculation of oestrone was 20% that of the first, and the Cmax of the third circulation was 50% that of the second. The areas under the serum-concentration-time curves (AUC) for the second and third recirculations were similar-each comprised 12-13% of the total AUC. The oral clearance values of the recirculations were constant (590Lh(-1)). Enterohepatic recirculation of endogenous compounds is aimed at maintaining a steady-state serum concentration for immediate use and hydrolysis in the target organs. It is concluded that exogenously added oestradiol and its metabolites follow the recirculation pathways of the endogenous oestrogen pool. PMID- 9751450 TI - Relative dispersions of intra-albumin transit times across rat and elasmobranch perfused livers, and implications for intra- and inter-species scaling of hepatic clearance using microsomal data. AB - It is recognized that vascular dispersion in the liver is a determinant of high first-pass extraction of solutes by that organ. Such dispersion is also required for translation of in-vitro microsomal activity into in-vivo predictions of hepatic extraction for any solute. We therefore investigated the relative dispersion of albumin transit times (CV2) in the livers of adult and weanling rats and in elasmobranch livers. The mean and normalized variance of the hepatic transit time distribution of albumin was estimated using parametric non-linear regression (with a correction for catheter influence) after an impulse (bolus) input of labelled albumin into a single-pass liver perfusion. The mean+/-s.e. of CV2 for albumin determined in each of the liver groups were 0.85+/-0.20 (n = 12), 1.48+/-0.33 (n = 7) and 0.90+/-0.18 (n = 4) for the livers of adult and weanling rats and elasmobranch livers, respectively. These CV2 are comparable with that reported previously for the dog and suggest that the CV2 of the liver is of a similar order of magnitude irrespective of the age and morphological development of the species. It might, therefore, be justified, in the absence of other information, to predict the hepatic clearances and availabilities of highly extracted solutes by scaling within and between species livers using hepatic elimination models such as the dispersion model with a CV2 of approximately unity. PMID- 9751451 TI - Time-dependent effects of Klebsiella pneumoniae endotoxin on hepatic drug metabolizing enzyme activity in rats. AB - The time-dependent effects of Klebsiella pneumoniae endotoxin on hepatic cytochrome P450-dependent drug-metabolizing capacity (cytochrome P450 and b5 content, activity of aminopyrine N-demethylase, p-nitroanisole O-demethylase, aniline hydroxylase and benzphetamine N-demethylase) and on the pharmacokinetics of antipyrine have been determined in rats. Measurement of enzyme activity and antipyrine (after intravenous injection of 20 mg kg(-1)) were performed 2, 24 and 96 h after a single intraperitoneal injection of endotoxin (1 mg kg(-1)) and after repeated doses (once daily for 4 days). The contribution of tumour necrosis factor alpha (TNFalpha) to the endotoxin-induced changes was also examined in rats pretreated with granulocyte colony-stimulating factor (G-CSF). The systemic clearance of antipyrine and the activity of hepatic cytochrome P450-dependent drug-metabolizing enzymes were dramatically reduced 24 h after a single injection of endotoxin, but had returned to control levels by 96h. The magnitudes of these decreases in these measurements after repeated doses of endotoxin were similar to those seen 24h after the single dose. The systemic clearance of antipyrine correlated significantly with cytochrome P450 content and aminopyrine N demethylase activity. In histopathological experiments, moderate hypertrophy of Kupffer cells was observed, with no evidence of severe liver-tissue damage. G-CSF pretreatment suppressed the increased plasma concentrations of TNFalpha produced 2 h after single endotoxin injection, but did not eliminate the endotoxin-induced decrease in the systemic clearance of antipyrine, suggesting that TNFalpha is not the sole component responsible for the reduction of cytochrome P450-mediated drug metabolizing enzyme activity. These results provide evidence that a single intraperitoneal injection of 1.0 mgkg(-1)K. pneumoniae endotoxin in rats reduces hepatic P450 and b5 levels, and reduces the activity of various cytochrome P450 mediated drug-metabolizing enzymes without causing severe liver-tissue damage. This suggests that the effect of endotoxin on hepatic cytochrome P450-mediated drug-metabolizing isozymes is non-selective. PMID- 9751452 TI - Effects of tetramethylpyrazine on portal hypertensive rats. AB - The effects of tetramethylpyrazine, an alkaloid isolated from a Chinese herb Ligusticum wallichii Franch have been assessed in portal hypertensive rats. Portal hypertension was induced by partial portal vein ligation in Sprague-Dawley rats. Two weeks after ligation, when the hyperdynamic state had stabilized, rats were anaesthetized after an overnight fast and cannulated for measurement of mean arterial pressure, portal venous pressure, cardiac index and heart rate. Tetramethylpyrazine (3.0, 9.9 and 30mgkg(-1)) induced dose-dependent reductions of portal venous pressure and mean arterial pressure after intravenous infusion. The maximum percentage reduction of portal venous pressure after tetramethylpyrazine was 6.0+/-0.8, 9.3+/-1.6 and 20+/-2% of baseline for doses of 3.0, 9.9 and 30.0mgkg(-1), respectively. Also, total peripheral resistance was significantly reduced by tetramethylpyrazine and cardiac index was slightly increased. Our results showed that tetramethylpyrazine induced portal pressure reduction in portal hypertensive rats. PMID- 9751453 TI - Endothelium-dependent relaxation in response to ethanol in the porcine isolated pulmonary artery. AB - Many drugs cannot be dissolved in distilled water and so other solvents such as ethanol, dimethylsulphoxide and methanol are used. Because very little is known about the direct effects of these three solvents on the cardiovascular system, we have examined their effects on isolated pulmonary and coronary arteries from the pig. Increasing concentrations of ethanol, dimethylsulphoxide and methanol induced relaxation in porcine pulmonary (at 1.2% v/v, 59.9+/-9.0% (n =9), 55.9+/ 9.0% (n =6) and 12.3+/-6.4% (n = 8), respectively, of U46619-induced tone) and coronary arteries (at 1.2% v/v, 69.9+/-7.1% (n = 10), 78.9+/-6.1% (n = 7) and 12.9+/-8.2% (n = 6) respectively, of U46619-induced tone). In the pulmonary arteries the relaxation in response to ethanol was found to be endothelium dependent whereas the responses to dimethylsulphoxide and methanol were unaffected by removal of the endothelium. In the coronary arteries the relaxation to all three solvents was independent of the presence of the endothelium. Comparison of the sensitivity of the tissues to the solvents showed that ethanol and dimethylsulphoxide produced comparative responses in both the pulmonary and coronary arteries, whereas methanol was much less potent. The endothelium dependent response to ethanol in the porcine pulmonary artery (maximum response, Emax, 67.1+/-9.3% of U46619-induced tone, n = 7) was attenuated by the cyclooxygenase inhibitor, flurbiprofen (Emax 31.9 +/- 12.0%, n=7), the nitric oxide synthase inhibitor, L-NAME (NG-nitro-L-arginine methyl ester; Emax 23.5+/ 10.2%, n = 7)) and the combination of both inhibitors (Emax 18.3+/-7.8%, n = 7). The residual relaxatory response to ethanol was abolished, and converted into a contractile response, both by removal of the endothelium (at 1.7% v/v ethanol 27.3+/-11.5% of U46619-induced tone, n=7) and by the addition of a low concentration of KC1 (49.9-/+10.3%, n=6), suggesting the release of a non prostanoid, non-nitric oxide factor from the endothelium. This response, however, was not attenuated by the cannabinoid receptor-antagonist SR141716A (N-(piperidin 1-yl)-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-me thyl-1H-pyrazole-3 carboxamide HCL; 52.5-/+4.3% relaxation, n =8), suggesting that the factor released in this preparation by ethanol is not a cannabinoid. The results of this study indicate that many solvents commonly used in pharmacological experiments have pronounced vasoactive properties. Methanol might be the vehicle of choice, because it was the least active solvent, whereas high concentrations of ethanol might influence vascular function at both the level of the smooth muscle and the endothelium, with the action on the endothelium involving the release of endothelium-derived relaxing factors. PMID- 9751454 TI - Delayed treatment with YM90K, an AMPA receptor antagonist, protects against ischaemic damage after middle cerebral artery occlusion in rats. AB - The neuroprotective effect of an alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionate (AMPA) receptor antagonist YM90K [6-(1H-imidazol-1-yl)-7 nitro-2,3(1H,4H)-quinoxalinedione monohydrochloride] has been examined in a rat middle cerebral artery occlusion model. Intravenous infusion of YM90K (2.5 20mgkg(-l)h(-l) for 4h) starting immediately after occlusion of the middle cerebral artery significantly reduced the cortical infarct volume 24h after occlusion compared with the control group. The protection at the highest dose was 39% (P < 0.05). Similar protective effects were observed when YM90K (20mgkg(-1)h( 1) for 4h) was delayed up to 2h after middle cerebral artery occlusion (45% reduction, P < 0.05). CNS1102 [N-(1-naphthyl)-N'-(3-ethylphenyl)-N' methylguanidine hydrochloride], a non-competitive N-methyl-D-aspartate (NMDA) receptor antagonist, also reduced the cortical infarct volume when 1.13mgkg(-1) was administered by intravenous bolus injection immediately after middle cerebral artery occlusion, followed by intravenous infusion at 0.785mgkg(-l)h(-1) for 4h (35% reduction, P<0.05). This neuroprotective effect was not observed when administration was delayed lh after middle cerebral artery occlusion. These results suggest that AMPA receptors might play a more important role than NMDA receptors in the late development of neuronal cell damage after focal cerebral ischaemia and that AMPA receptor blockade would be one beneficial strategy in treating acute stroke. PMID- 9751455 TI - Prejunctional control of pH 6-induced bronchoconstriction by NK1, NK2, mu-opioid, alpha2-adrenoceptor and glucocorticoid receptors in guinea-pig isolated perfused lung. AB - This study investigated the release of calcitonin-gene related peptide-like (CGRP) immunoreactivity and bronchoconstriction induced by pH 6 buffer in guinea pig isolated perfused lung. Both pH 6-induced CGRP-like immunoreactivity and bronchoconstriction were completely abolished after systemic pretreatment with capsaicin. Pretreatment with the NK2 receptor antagonist SR 48968 (5 x 10(-7)M) completely inhibited bronchoconstriction and significantly reduced the immunoreactivity induced by the pH 6 buffer. The NK1 antagonist SR 140333 (5 x 10(-7)M) and, to a lesser extent the NK1 antagonist CP 96345, morphine (5 x 10( 6)M), the alpha2-adrenoceptor agonist UK 14304 (10(-7)M) and betamethasone (10( 6)M) significantly reduced both pH 6-induced bronchial response and CGRP-like immunoreactivity overflow. The effects of morphine and UK14304 were partially reversed by naloxone (5 x 10(-5)M) and idazoxan (5 x 10(-50M). Therefore, NK1, NK2, mu-opioid, alpha2-adrenoceptor and glucocorticoid receptors seemed to have a prejunctional action on pH 6 buffer-induced CGRP-like immunoreactivity and bronchoconstriction. PMID- 9751456 TI - Effect of different beta-adrenergic agonists on the intestinal absorption of galactose and phenylalanine. AB - Nutrient transport across the mammalian small intestine is regulated by several factors, including intrinsic and extrinsic neural pathways, paracrine modulators, circulating hormones and luminal agents. Because beta-adrenoceptors seem to regulate gastrointestinal functions such as bicarbonate and acid secretion, intestinal motility and gastrointestinal mucosal blood flow, we have investigated the effects of different beta-adrenergic agonists on nutrient absorption by the rat jejunum in-vitro. When intestinal everted sacs were used the beta2-agonist salbutamol had no effect either on galactose uptake by the tissue or mucosal-to serosal flux whereas mixed beta1- and beta2-agonists (isoproterenol and orciprenaline) and beta3-agonists (BRL 35135, Trecadrine, ICI 198157 and ZD 7114) inhibited galactose uptake and transfer of D-galactose from the mucosal-to serosal media across the intestinal wall (although the inhibiting effects of isoproterenol and Trecadrine were not statistically significant). In intestinal everted rings both Trecadrine and BRL 35135 clearly reduced galactose uptake, the effect being a result of inhibition of the phlorizin-sensitive component. Total uptake of phenylalanine by the intestinal rings was also reduced by those beta3 adrenergic agonists. These results suggest that beta1- and beta3-adrenergic receptors could be involved in the regulation of intestinal active transport of sugars and amino acids. PMID- 9751457 TI - Use of protease inhibitors to improve calcitonin absorption from the small and large intestine in rats. AB - The objective of this study was to examine the effects of protease inhibitors on the absorption of calcitonin from different regions of the intestine in rats. The absorption experiments were investigated by in-situ use of closed intestinal loops in rats and stability of calcitonin was examined in mucosal homogenates and intestinal fluids. The intestinal absorption of calcitonin was evaluated by measurement of its hypocalcaemic effect. No substantial hypocalcaemic response was observed when calcitonin was administered into the jejunum or colon. A slight hypocalcaemic effect was observed after administration of calcitonin into the ileum. Of the co-administered protease inhibitors, bacitracin (20mM) strongly promoted calcitonin absorption from the jejunum, ileum and colon. A significant hypocalcaemic effect was also obtained after intestinal administration of calcitonin with soybean trypsin inhibitor (10mgmL(-1)), camostat mesylate (20mM) or aprotinin (2mgmL(-1)). In the stability experiment, bacitracin reduced the degradation of calcitonin in the different intestinal homogenates. Soybean trypsin inhibitor significantly reduced the degradation of calcitonin in the fluids of the small intestine. We also examined the different endopeptidases in gut luminal fluids and the different exopeptidases in gut mucosal homogenates of rats. The ranking order for the total endopeptidase activity of the intestinal fluids was jejunum > ileum > colon. That for total exopeptidase activity of the intestinal mucosa was jejunum > ileum > colon. These results suggest that endo- and exopeptidases might be responsible for the hydrolysis of calcitonin and that protease inhibitors might usefully improve absorption of calcitonin to the systemic circulation from the large intestine. PMID- 9751458 TI - Influence of trimebutine on inflammation- and stress-induced hyperalgesia to rectal distension in rats. AB - The effects of trimebutine and its major metabolite, N-desmethyltrimebutine on inflammation- and stress-induced rectal hyperalgesia have been evaluated in rats fitted with electrodes implanted in the longitudinal striated muscle of the abdomen. Intermittent rectal distension was performed before and 3 days after induction of rectal inflammation by local infusion of trinitrobenzenesulphonic acid (in ethanol). Stress consisted of 2h partial restraint and rectal distension was performed before and 30min after the end of the partial restraint session. The animals were treated intraperitoneally with trimebutine or desmethyltrimebutine (5, 10 or 20mgkg(-1)) or vehicle 15min before rectal distension. Naloxone (1mgkg(-1)) or saline was injected subcutaneously before trimebutine and desmethyltrimebutine. Before treatment trimebutine at the highest dose (20mgkg(-1)) reduced the abdominal response to rectal distension for the highest volume of distension (1.6mL) whereas desmethyltrimebutine was inactive. After rectocolitis the abdominal response to rectal distension was enhanced and trimebutine at 5mgkg(-1) reduced and at 10 mgkg(-1) suppressed inflammation induced hyperalgesia, an effect reversed by naloxone. Desmethyltrimebutine was inactive. Stress-induced hypersensitivity was attenuated or suppressed, or both, by trimebutine and desmethyltrimebutine at doses of 5, 10 or 20mgkg(-l); greater efficacy was observed for desmethyltrimebutine and the effects were not reversed by naloxone. It was concluded that trimebutine and desmethyltrimebutine are active against inflammation- and stress-induced rectal hyperalgesia but act differently. The effect of trimebutine on inflammation-induced hyperalgesia is mediated through opioid receptors. PMID- 9751459 TI - Nicotine-induced perturbations on heart rate, body temperature and locomotor activity daily rhythms in rats. AB - The aim of this study was to evaluate the influence of nicotine on the daily rhythms of heart rate, body temperature and locomotor activity in unrestrained rats by use of implanted radiotelemetry transmitters. The study was divided into three seven-day periods: a control period, a treatment period and a recovery period. The control period was used for baseline measurement of heart rate, body temperature and locomotor activity. During the treatment period three rats received nicotine (1 mg kg(-1), s.c.) at 0900 h. Three rats received saline under the same experimental conditions. Heart rate, body temperature and locomotor activity were continuously monitored and plotted every 10 min. During the three periods a power spectrum analysis was used to determine the dominant period of rhythmicity. If daily rhythms of heart rate, body temperature and locomotor activity were detected, the characteristics of these rhythms, i.e. the mesors, amplitudes and acrophases, were determined by cosinor analysis, expressed as means +/- s.e.m. and compared by analysis of variance. Nicotine did not suppress daily rhythmicity but induced decreases of amplitudes and phase-advances of acrophases for heart rate, body temperature and locomotor activity. These perturbations might result from the effects of nicotine on the suprachiasmatic nucleus, the hypothalamic clock that co-ordinates biological rhythms. PMID- 9751460 TI - Influence of twinline, an elemental diet, on the generation of nitric oxide and reactive-oxygen intermediates from mouse peritoneal macrophages and polymorphonuclear leukocytes. AB - The influence of Twinline (SNN-6010), an elemental diet containing medium-chain triglycerides, on the generation of nitric oxide (NO) and superoxide (O2.-) has been examined in mouse peritoneal macrophages and polymorphonuclear leukocytes (PMN). When PMN and peritoneal macrophages obtained from untreated mice were cultured in medium containing 0.1% and 1% (v/v) Twinline for 48h and stimulated with phorbol myristate acetate or N-formyl-methionyl-leucyl-phenylalanine, their chemiluminescence and O2.- generation were strongly suppressed, as was NO generation from peritoneal macrophages. PMN and peritoneal macrophages obtained from mice fed Twinline for 30 days generated much smaller amounts of 02.- and NO compared with PMN and peritoneal macrophages from control mice. In conjunction with this suppressed NO generation, inducible NO synthase and its mRNA expression in peritoneal macrophages were suppressed by Twinline both in-vivo and ex-vivo. Although phagocytosis of PMN and peritoneal macrophages was not suppressed by Twinline; their candida-killing activity was markedly suppressed. These results indicate that Twinline suppresses the host-defence function of PMN and peritoneal macrophages by down-regulating their generation of reactive-oxygen intermediates and NO. PMID- 9751461 TI - Delayed healing of chronic gastric ulcer after Helicobacter pylori infection in mice. AB - It has been suggested that there is a close relationship between Helicobacter pylori and the onset or recurrence of gastroduodenal disease. The aim of this study was to examine the effect of H. pylori on the healing of chronic gastric ulcers induced in mice. H. pylori administered to nude mice delayed the healing of experimental acetic acid-induced gastric ulcers. Histological examination showed the occurrence of high densities of H. pylori on the surface of epithelial cells and in the ulcerated area. Repeated administration of amoxicillin (10 mgkg( 1) daily for 5 days) eradicated H. pylori and increased the rate of healing of gastric ulcers in H. pylori-infected mice, but metronidazole, which also eradicated the organisms, did not significantly affect the rate of healing. In conclusion, H. pylori-infection delayed the healing of gastric ulcers induced by the serosal application of acetic acid in mice, possibly by aggravation or prolongation of the mucosal inflammation. Amoxicillin eradicated H. pylori and promoted gastric ulcer healing in mice infected with H. pylori. PMID- 9751462 TI - A comparative study of the effects of sparteine, lupanine and lupin extract on the central nervous system of the mouse. AB - Lupin is toxic because of its alkaloid content, sparteine and lupanine in particular. Although the pharmacological properties of sparteine are well known those of lupanine have not been much studied. This paper reports procedures for extraction, purification and crystallization of lupanine, and methods for the preparation of an extract for injection of Lupinus mutabilis Sweet, and for the determination of the acute toxicity and maximum non-lethal dose (DL0) of lupanine, sparteine and lupin extract in the mouse. The three substances were tested on the central nervous system (CNS) for locomotor activity, for interaction with specific drugs used for treatment of the CNS (the stimulant drugs amphetamine and pentetrazol and the depressant drugs pentobarbital and chlorpromazine) and for analgesic activity. The results indicate that lupanine and lupin extract are less toxic than sparteine and that at the doses studied the three products have a weak sedative effect on the CNS. PMID- 9751463 TI - Systemic glucocorticoid treatment in rheumatoid arthritis--a debate. AB - Arguments are presented for and against the use of oral glucocorticoid treatment in patients with rheumatoid arthritis. Controlled clinical trials, uncontrolled longitudinal observations and accumulated clinical experience are drawn together to place in perspective treatment decisions in routine clinical practice. The evidence points to a relatively short term improvement in symptom control and a longer term benefit in reducing progressive joint destruction, but to this must be added fears about inappropriate prescription of glucocorticoids with consequent adverse effects. Areas requiring further research are highlighted. PMID- 9751464 TI - Correlation of serum IgA rheumatoid factor levels with disease severity in rheumatoid arthritis. AB - In the present study, the IgA rheumatoid factor (IgA RF) was assayed by ELISA in 114 sera of patients with active RA. IgA RF was found in the sera of 38 (33.3%) patients. A total of 56 disease variables were studied, differences in variables between IgA RF positive and negative patients were analyzed. The presence of IgA RF was found to be associated with a rapid progressive course, RA activity, onset of extra-articular manifestations (especially systemic rheumatoid vasculitis), high IgM RF titres, circulating immune complexes, cryoglobulins, and C-reactive protein levels. There was no statistically significant difference in serum IgA levels between IgA RF seropositive and seronegative patients. The presence of IgA RF showed a positive correlation with the level of IgM RF. The results of this study showed that the determination of IgA RF has a practical use in the establishment of the severe course of RA with systemic damages. PMID- 9751466 TI - Patient and parent experiences with health care services in pediatric rheumatology. AB - The health care received from first admission to a pediatric rheumatology clinic to 9-year follow-up was assessed in 109 patients with chronic inflammatory rheumatic diseases or chronic idiopathic musculoskeletal pain. Ninety-five of the patients had received hospital care after the first admission, of whom 53 patients > or = 18 years, 21 patients < 18 years, and 33 parents of patients < 18 years rated their degree of satisfaction with the health care from 0 to 10. Mean scores of satisfaction with different aspects of care ranged from 6.0 to 9.6. Among patients > or = 18 years, those with idiopathic pain were less satisfied than those with inflammatory rheumatic diseases on the availability of care (mean 6.4 vs. 8.5, p < 0.001), continuity of care (mean 6.5 vs. 8.4, p < 0.001), and empathy of the health care providers (mean 6.7 vs. 7.9, p <0.05). The diagnostic group and the occurrence of remission predicted the level of global satisfaction in patients > or = 18 years. In patients < 18 years, chronic family difficulties predicted patient satisfaction and physical disability and chronic family difficulties predicted parent satisfaction. In conclusion, most parents and patients with inflammatory rheumatic diseases were satisfied with the health care. However, some patients with idiopathic pain had unmet needs for care. PMID- 9751465 TI - Chronic arthritis and gamma heavy chain disease: coincidence or pathogenic link? AB - In 1991, gamma heavy chain disease was diagnosed in a 43-year-old female, who 3 years earlier had contracted an erosive seronegative chronic arthropathy. Her gamma heavy chain disease had a benign course, requiring no specific therapy for 5 years. In 1996, however, her lymphoproliferative disorder underwent a more malignant course, with renal and cardiac failure and increasing articular problems, requiring treatment with melphalan and prednisolone, following the protocol for myelomatosis. Laboratory studies revealed a monoclonal component in serum and urine. consistent with dimers of gamma-chains of the gamma3 subclass, but with a smaller molecular mass than normal gamma3-chains, suggesting molecular aberrations as consistently observed in this disorder. Massive localization of plasma cells and blasts with cytoplasmic or cell membrane staining for gamma3 chains, but no staining for kappa or lambda light chains, was observed by immunohistochemical studies of tissue specimens from bone marrow as well as affected synovial tissue. Large amounts of extracellular gamma3-chains were deposited in the synovial membrane. In addition, marked inflammatory changes with synovial cell hyperplasia were seen. Whether the present case represents primarily a gamma heavy chain deposition disease with reactive inflammatory changes in the joints, or another example of gamma heavy chain disease preceded by seronegative rheumatoid arthritis, remains elusive. Regardless, a possible pathogenic link between the two disease processes is an intriguing possibility. PMID- 9751467 TI - Self-reported bodily pain in schoolchildren. AB - It has been suggested that musculoskeletal symptoms develop from early age and can be regarded as a lifespan phenomenon. The study of childhood pain might provide a better understanding of the origin of chronic pain in adults. In a study of 569 schoolchildren, aged 10-15 years, in a local community close to Oslo, 75% reported that they usually experience bodily pain. Girls reported more pain than boys. 25% of those reporting pain experience symptoms several days a week. Knee symptoms and back pain were most frequently reported. Thirty-seven % of the girls reported headache, only 20% of the boys. Girls also reported more neck and shoulder pain than boys. The oldest respondents reported symptoms from more body parts. Symptoms from several body parts were more frequent among girls. Thirty-eight % of the respondents reported that it sometimes is hard to concentrate because of the pain, and 26% reported that they sometimes have to use medication. The consequences of pain increased with increasing age and increasing number of body parts affected. The results are consistent with findings in the adult population. PMID- 9751468 TI - Effects of intravenous regional administration of methylprednisolone plus mepivacaine in rheumatoid arthritis. AB - To evaluate the subjective and objective response to intravenous regional administration of a glucocorticoid (IVRAG) on rheumatoid arthritis (RA), twenty RA-patients received, in a randomised, double-blind crossover and placebo controlled fashion, either 50 mg methylprednisolone in mepivacaine 0.25% or mepivacaine (placebo) in one hand only using a Bier-block technique. The other hand was given the opposite. One week later the procedure was repeated but the previous placebo hand was now treated with the glucocorticoid. About 50% of the patients experienced a subjective improvement at 1 and 6 weeks. After one week a significant reduction was recorded in grip diastasis with no difference between the glucocorticoid and the placebo. In the other outcome measures (grip strength, handvolume, rest pain, and movement provoked pain) no differences were recorded between the glucocorticoid and placebo. At six weeks a significant reduction in grip diastasis and movement provoked pain as well as a significant increase in grip strength were noted. Hand volume was unchanged. IVRAG is a safe, easy and well-tolerated technique. The beneficial results are probably due to both a systemic and a regional effect. The findings of benefit in both hands at the end of one week seem to suggest that mepivacaine can give some short term improvement. PMID- 9751469 TI - Effects of static and dynamic shoulder rotator exercises in women with rheumatoid arthritis: a randomised comparison of impairment, disability, handicap, and health. AB - The aim was to compare static and dynamic shoulder rotator endurance training in a group of women with mild rheumatoid arthritis and to see whether such training could influence impairment, disability, and handicap. The effects on general health were also studied. Patients were randomly assigned to a static (n = 17) (average age 59, median disease duration 7) or a dynamic training group (n = 20) (average age 56, median disease duration 10.5). Measurements were taken at the start, 10 weeks later when the training period was finished, and after a further 10 weeks. After the training both groups had fewer swollen joints in the upper extremity and less shoulder-arm pain. The dynamic group patients also improved according to the physical and overall dimensions of the Sickness Impact Profile. As impairment and aspects of disability and handicap were influenced by training but not by the patients' opinions regarding perceived disease activity and health, these relationships must be studied further. PMID- 9751470 TI - Clinical experience of 13 cases with severe pulmonary hemorrhage in systemic lupus erythematosus with active nephritis. AB - Pulmonary hemorrhage is a rare, but serious manifestation of systemic lupus erythematosus (SLE). Herein, we report 13 cases of severe pulmonary hemorrhage in SLE. Hemoptysis was present in 11 patients. All thirteen patients had active nephritis and were in the stage of nephrotic syndrome. A majority of the patients had neuropsychiatric manifestations and coagulopathy including thrombocytopenia or lupus anticoagulant. All episodes of pulmonary hemorrhage occurred after large dose of corticosteroid had been administered in treating nephritis. Recurrent pulmonary hemorrhage was noted in four patients. Ten (77%) of the 13 patients finally died. Respiratory failure was the main cause of death. Our observation suggests that active nephritis with hypoalbuminemia is a major risk factor for severe pulmonary hemorrhage in SLE patients and that high dose corticosteroid use can not prevent the occurrence of severe pulmonary hemorrhage in SLE. PMID- 9751471 TI - Soluble cell adhesion molecules--P-selectin and ICAM-1, and disease activity in patients receiving sulphasalazine for active rheumatoid arthritis. AB - The aim of this pilot study was to examine soluble cell adhesion molecules before and after sulphasalazine (SSZ) therapy in active RA. Assessment of RA patients (n = 13) was undertaken before and after 3 months of SSZ. sICAM-1, sVCAM-1, sP- and sE-selectin were measured using an ELISA. The mean (+/-SEM) C-reactive protein (CRP) and sP-selectin levels were significantly reduced from 3.9(0.89) to 2.01(0.53) mg/dl and from 332.8 (48.2) to 116.2 (11.1) respectively, after 3 months of SSZ. The sICAM-1 and sP-selectin levels were significantly higher in RA patients at baseline and a reduction occurred of sICAM-1, sVCAM-1 and sE-selectin levels, however this was not significant. The fall in mean (SEM) sICAM-1, from 345.0 (29.8) to 333.5 (30.2), correlated with the change in CRP (r=0.66; p = 0.018), but the fall in sP-selectin did not. SSZ therapy reduced sP-selectin and sICAM-1 levels in active RA, sICAM-1 correlates with disease activity. SSZ may reduce platelet and/or endothelial activity in RA which may be a useful marker of response, however studies of longer duration and more patients are required. PMID- 9751472 TI - Early referral and outcome in rheumatoid arthritis. AB - In a cross-sectional study of 200 outpatients with rheumatoid arthritis (RA) we tested the hypothesis that early specialist referral within one year of disease onset reduces subsequent functional disability. Early referral was defined as being seen in a specialist unit within one year of the onset of symptoms. Functional outcome was measured using the physical functional section of the full test (NHP) and (HAQ). Additional information was collected on other factors likely to influence disability including age, sex, pain scores, and rheumatoid factor positivity. The difference in average HAQ and NHP physical function scores between late and early referrals was 0.34 and 11.0 respectively (p< 0.01 in both cases) using unequal-variance t-test. After adjusting for the other risk factors, multiple regression analysis showed late referral patients had greater NHP physical function scores than early-referred patients by approximately 8 points and there was a similar trend with HAQ. We conclude that early referral for specialist advice is associated with improved health status, with reduced physical function scores on the NHP. PMID- 9751474 TI - Coexisting psoriatic arthritis, gout, and chondrocalcinosis. AB - The authors describe the case of a 63-year-old Caucasian man with a history of psoriatic arthritis who subsequently developed gout and chondrocalcinosis. This is the first report documenting the simultaneous occurrence of psoriatic arthritis, gout, and chondrocalcinosis in a single patient. The relations of these rheumatic diseases are discussed. PMID- 9751473 TI - Suboptimal clinical response to anti-tumor necrosis factor alpha (TNFalpha) antibody therapy in a patient with severe rheumatoid arthritis and lymphadenopathy. AB - This concerns a patient with severe rheumatoid arthritis (RA) and lymphadenopathy (LA) who showed suboptimal clinical response to antitumor necrosis factor alpha (TNFalpha) antibody (Ab), cA2 therapy. The assessment of TNFalpha and IL-6 mRNA expression in the swollen lymph-node (LN) of the patient by reverse transcription, polymerase chain reaction (RT-PCR) before cA2 treatment, showed only enhanced IL-6 production, but not TNFalpha. Moreover, cA2 failed to inhibit in-vitro spontaneous IL-6 production in the LN block culture from the patient. Taken together, these results indicate that IL-6 production in the swollen LNs of the patient might not depend on TNFalpha. This might partly cause suboptimal clinical response to anti-TNFalpha Ab therapy in the patient. PMID- 9751475 TI - Guillain-Barre syndrome in a child with Henoch-Schonlein Purpura. AB - This report details a 10 year old girl with Henoch-Schonlein Purpura (HSP) whose primary illness was subsequently complicated by the development of the Guillain Barre syndrome. This association is of extremely rare occurrence. PMID- 9751476 TI - Arteriovenous malformation of knee masquerading as juvenile arthritis. AB - This is a rare case of involvement of knee joint by an arteriovenous malformation. A nine year old girl had recurrent monoarticular pain and swelling of long duration and had been treated as juvenile rheumatoid arthritis. Angiographic and histological features suggested an arteriovenous malformation of lower thigh with knee joint involvement. PMID- 9751477 TI - Reiter's syndrome induced by Gardnerella vaginalis. PMID- 9751478 TI - More pieces of the puzzle in place, even more discovered missing. PMID- 9751479 TI - Recommendations for nomenclature of the insulin-like growth factor binding protein superfamily. PMID- 9751480 TI - Muscarinic regulation of intracellular signaling and neurosecretion in gonadotropin-releasing hormone neurons. AB - Agonist activation of cholinergic receptors expressed in perifused hypothalamic and immortalized GnRH-producing (GT1-7) cells induced prominent peaks in GnRH release, each followed by a rapid decrease, a transient plateau, and a decline to below basal levels. The complex profile of GnRH release suggested that acetylcholine (ACh) acts through different cholinergic receptor subtypes to exert stimulatory and inhibitory effects on GnRH release. Whereas activation of nicotinic receptors caused a transient increase in GnRH release, activation of muscarinic receptors inhibited basal GnRH release. Nanomolar concentrations of ACh caused dose-dependent inhibition of cAMP production that was prevented by pertussis toxin (PTX), consistent with the activation of a plasma-membrane Gi protein. Micromolar concentrations of ACh also caused an increase in phosphoinositide hydrolysis that was inhibited by the M1 receptor antagonist, pirenzepine. In ACh-treated cells, immunoblot analysis revealed that membrane associated G(alpha q/11) immunoreactivity was decreased after 5 min but was restored at later times. In contrast, immunoreactive G(alpha i3) was decreased for up to 120 min after ACh treatment. The agonist-induced changes in G protein alpha-subunits liberated during activation of muscarinic receptors were correlated with regulation of their respective transduction pathways. These results indicate that ACh modulates GnRH release from hypothalamic neurons through both M1 and M2 muscarinic receptors. These receptor subtypes are coupled to Gq and Gi proteins that respectively influence the activities of PLC and adenylyl cyclase/ion channels, with consequent effects on neurosecretion. PMID- 9751481 TI - The pulsatile characteristics of hypothalamo-pituitary-adrenal activity in female Lewis and Fischer 344 rats and its relationship to differential stress responses. AB - The dynamic patterns of basal and stimulated hypothalamo-pituitary-adrenal (HPA) activity of freely moving female Lewis and Fischer 344 rats were compared using an automated blood-sampling system. Both strains showed pulsatile corticosterone release throughout the 24 h cycle. Lewis rats showed clear circadian variation in both pulse frequency (8.4 +/- 0.4 pulses between 1700-2300 h vs. 5.3 +/- 0.8 pulses between 0500-1100 h; P < 0.05) and height (198 +/- 27 ng/ml between 1700 2300 h vs. 107 +/- 14 ng/ml between 0500-1100 h; P < 0.05). Fischer rats exhibited pulses of similar frequency and height to those in Lewis rats during the evening, but showed no circadian variation, resulting in higher mean daily corticosterone concentrations. Although both strains showed behavioral and HPA responses to white noise stress (10 min; 114 dB), Fischer rats showed much greater increases in total activity, grooming, and rearings, and two important differences in the corticosterone responses were observed. First, in Lewis rats a clear relationship existed between basal and stimulated HPA activities, in that a significant response was seen only when the stress coincided with the rising (secretory active) phase of a basal pulse. Noise stress coinciding with a falling (nonsecretory) phase elicited no significant response. In contrast, Fischer rats showed similar responses regardless of the underlying pulse phase. Second, after the peak response at 20 min (Lewis, 237 +/- 67 ng/ml; Fischer, 390 +/- 57 ng/ml), corticosterone levels fell rapidly in Lewis rats, but remained maximally elevated for 20 min in Fischer rats, resulting in a significantly greater integrated response. The corticosterone response to i.v. CRF was unaffected by pulse phase in both strains, suggesting that a suprapituitary mechanism mediates the phase dependent response to stress in the Lewis strain. CRF-induced corticosterone levels rose more rapidly in Fischer rats, peaking at 10 min (473 +/- 95 ng/ml) compared with 30 min (390 +/- 75 ng/ml) in Lewis rats, suggesting greater pituitary sensitivity in this strain. Thus, differences in both central and pituitary control of the HPA axis contribute to the strain difference in stress responsiveness between female Lewis and Fischer rats. PMID- 9751482 TI - Nitric oxide inhibits aldosterone synthesis by a guanylyl cyclase-independent effect. AB - To investigate the mechanism of nitric oxide (NO) inhibition of aldosterone release, this study compared the effects of type A natriuretic peptide and heat stable enterotoxin to a nitric oxide donor, deta nonoate, on cGMP production and angiotensin II-stimulated aldosterone synthesis ill primary cultures of bovine adrenal zona glomerulosa cells. Type A natriuretic peptide (10(-10)-10(-6) M) and deta nonoate (10(-6)-10(-3) M) stimulated concentration-related increases in cGMP production. Heat-stable enterotoxin (10(-6) M) failed to stimulate cGMP synthesis in zona glomerulosa cells. Type A natriuretic peptide and deta nonoate attenuated angiotensin II-stimulated aldosterone production over the same concentration range that stimulated cGMP production. Heat-stable enterotoxin (10(-6) M) was without effect on aldosterone release. To further test the hypothesis that cGMP mediated the inhibition of aldosterone synthesis, the selective inhibitor of soluble guanylyl cyclase, 1H-(1,2,4)oxadiazolo [4,3-a]quinoxalin-1-one (ODQ) was used. ODQ pretreatment (10(-5) M) completely prevented deta nonoate-stimulated cGMP production without altering the inhibitory effect of deta nonoate on angiotensin II-stimulated steroidogenesis. Consistent with its selectivity for inhibiting soluble guanylyl cyclase, ODQ did not block type A natriuretic peptide stimulated cGMP synthesis or type A natriuretic peptide inhibition of steroidogenesis. Deta nonoate completely blocked 25-hydroxycholesterol- and progesterone-stimulated aldosterone synthesis in zona glomerulosa cells and inhibited the conversion of 25-hydroxycholesterol to pregnenolone in mitochondrial fractions from bovine adrenal cortex. Deta nonoate-derived NO gave an absorbance maximum of the mitochondrial cytochrome P450 of 453 nm and inhibited the absorbance at 450 nm caused by carbon monoxide binding to the enzyme. These results suggest that deta nonoate reduces steroidogenesis independent of guanylyl cyclase activation and that NO has a direct effect to inhibit the activity of cytochrome P450, probably by binding to the heme groups of the cytochrome. PMID- 9751483 TI - Regulation of multicatalytic enzyme activity by insulin and the insulin-degrading enzyme. AB - The insulin-degrading enzyme (IDE) plays an important role in the cellular metabolism of insulin. Recent studies have also suggested a regulatory role for this protein in controlling the activity of cytoplasmic protein complexes, including the proteasome [multicatalytic proteinase (MCP)] and the glucocorticoid and androgen receptors. Binding of IDE to these complexes increases their activity, whereas the addition of substrates for IDE inhibits activity. This provides a potential mechanism of action for internalized insulin and other IDE substrates in the control of protein turnover. To examine further the interactions, partially purified IDE-MCP complex was treated with EDTA or EGTA, and activity was measured in the absence and presence of various divalent cations (Ca2+, Mn2+, Co2+, and Zn2+) and insulin. EDTA treatment reduced MCP activity and eliminated the effect of insulin on the complex. Divalent cations partially or completely restored MCP activity, but did not restore the effect of insulin. EGTA treatment had a lesser effect on MCP activity, but abolished insulin inhibition of activity. Divalent cations restored the insulin effect. Inhibitors of IDE also blocked the insulin effect on MCP activity, as did treatment with SDS. These findings suggest that conformational changes in the complex may play a role in the insulin control of MCP activity. PMID- 9751484 TI - Insulin potentiates platelet-derived growth factor action in vascular smooth muscle cells. AB - Correlative studies have indicated that hyperinsulinemia is present in many individuals with atherosclerosis. Insulin resistance has also been linked to cardiovascular disease. It has proved to be difficult to decipher whether hyperinsulinemia or insulin resistance plays the most important role in the pathogenesis of atherosclerosis and coronary artery disease. In this study, we demonstrate that insulin increases the amount of farnesylated p21Ras in vascular smooth muscle cells (VSMC), thereby augmenting the pool of cellular Ras available for activation by platelet-derived growth factor (PDGF). In VSMC incubated with insulin for 24 h, PDGF's influence on GTP-loading of Ras was significantly increased. Furthermore, in cells preincubated with insulin, PDGF increased thymidine incorporation by 96% as compared with a 44% increase in control cells (a 2-fold increment). Similarly, preincubation of VSMC with insulin increased the ability of PDGF to stimulate gene expression of vascular endothelial growth factor 5- to 8-fold. The potentiating influence of insulin on PDGF action was abrogated in the presence of a farnesyltransferase inhibitor. Thus, the detrimental influence of hyperinsulinemia on the arterial wall may be related to the ability of insulin to augment farnesyltransferase activity and provide greater amounts of farnesylated p21Ras for stimulation by various growth promoting agents. PMID- 9751485 TI - Mass spectrometric evidence that agents that cause loss of Ca2+ from intracellular compartments induce hydrolysis of arachidonic acid from pancreatic islet membrane phospholipids by a mechanism that does not require a rise in cytosolic Ca2+ concentration. AB - Stimulation of pancreatic islets with glucose induces phospholipid hydrolysis and accumulation of nonesterified arachidonic acid, which may amplify the glucose induced Ca2+ entry into islet beta-cells that triggers insulin secretion. Ca2+ loss from beta-cell intracellular compartments has been proposed to induce both Ca2+ entry and events dependent on arachidonate metabolism. We examine here effects of inducing Ca2+ loss from intracellular sequestration sites with ionophore A23187 and thapsigargin on arachidonate hydrolysis from islet phospholipids. A23187 induces a decline in islet arachidonate-containing phospholipids and release of nonesterified arachidonate. A23187-induced arachidonate release is of similar magnitude when islets are stimulated in Ca2+ replete or in Ca2+-free media or when islets loaded with the intracellular Ca2+ chelator BAPTA are stimulated in Ca2+-free medium, a condition in which A23187 induces no rise in beta-cell cytosolic [Ca2+]. Thapsigargin also induces islet arachidonate release under these conditions. A23187- or thapsigargin-induced arachidonate release is prevented by a bromoenol lactone (BEL) inhibitor of a beta-cell phospholipase A2 (iPLA2), which does not require Ca2+ for catalytic activity and which is negatively modulated by and physically interacts with calmodulin by Ca2+-dependent mechanisms. Agents that cause Ca2+ loss from islet intracellular compartments thus induce arachidonate hydrolysis from phospholipids by a BEL-sensitive mechanism that does not require a rise in cytosolic [Ca2+], and a BEL-sensitive enzyme-like iPLA2 or a related membranous activity may participate in sensing Ca2+ compartment content. PMID- 9751486 TI - Programmed administration of parathyroid hormone increases bone formation and reduces bone loss in hindlimb-unloaded ovariectomized rats. AB - Gonadal insufficiency and reduced mechanical usage are two important risk factors for osteoporosis. The beneficial effects of PTH therapy to reverse the estrogen deficiency-induced bone loss in the laboratory rat are well known, but the influence of mechanical usage in this response has not been established. In this study, the effects of programed administration of PTH on cancellous bone volume and turnover at the proximal tibial metaphysis were determined in hindlimb unloaded, ovariectomized (OVX), 3-month-old Sprague-Dawley rats. PTH was administered to weight-bearing and hindlimb-unloaded OVX rats with osmotic pumps programed to deliver 20 microg human PTH (approximately 80 microg/kg x day) during a daily 1-h infusion for 7 days. Compared with sham-operated rats, OVX increased longitudinal and radial bone growth, increased indexes of cancellous bone turnover, and resulted in net resorption of cancellous bone. Hindlimb unloading of OVX rats decreased longitudinal and radial bone growth, decreased osteoblast number, increased osteoclast number, and resulted in a further decrease in cancellous bone volume compared with those in weight-bearing OVX rats. Programed administration of PTH had no effect on either radial or longitudinal bone growth in weight-bearing and hindlimb-unloaded OVX rats. PTH treatment had dramatic effects on selected cancellous bone measurements; PTH maintained cancellous bone volume in OVX weight-bearing rats and greatly reduced cancellous bone loss in OVX hindlimb-unloaded rats. In the latter animals, PTH treatment prevented the hindlimb unloading-induced reduction in trabecular thickness, but the hormone was ineffective in preventing either the increase in osteoclast number or the loss of trabecular plates. Importantly, PTH treatment increased the retention of a baseline flurochrome label, osteoblast number, and bone formation in the proximal tibial metaphysis regardless of the level of mechanical usage. These findings demonstrate that programed administration of PTH is effective in increasing osteoblast number and bone formation and has beneficial effects on bone volume in the absence of weight-bearing and gonadal hormones. We conclude that the actions of PTH on cancellous bone are independent of the level of mechanical usage. PMID- 9751487 TI - Effects of castration and chronic steroid treatments on hypothalamic gonadotropin releasing hormone content and pituitary gonadotropins in male wild-type and estrogen receptor-alpha knockout mice. AB - Testicular androgens are integral components of the hormonal feedback loops that regulate circulating levels of LHbeta and FSH. The sites of feedback include hypothalamic areas regulating GnRH neurons and pituitary gonadotropes. To better define the roles of androgen receptor (AR), estrogen receptor-alpha (ERalpha), and estrogen receptor-beta (ERbeta) in mediating feedback effects of sex steroids on reproductive neuroendocrine function, we have determined the effects of castration and steroid replacement therapy on hypothalamic GnRH content, pituitary LHbeta and FSHbeta messenger RNA (mRNA) levels, and serum gonadotropins in male wild-type (WT) and estrogen receptor-alpha knockout (ERKO) mice. Hypothalami from intact WT and ERKO males contained similar amounts of GnRH, whereas castration significantly reduced GnRH contents in both genotypes. Replacement therapy with estradiol (E2), testosterone (T), or dihydrotestosterone (DHT) restored hypothalamic GnRH content in castrated (CAST) WT mice; only the androgens were effective in CAST ERKOs. Analyses of pituitary function revealed that LHbeta mRNA and serum LHbeta levels in intact ERKOs were 2-fold higher than those in intact WT males. Castration increased levels of LHbeta mRNA (1.5- to 2 fold) and serum LHbeta (4- to 5-fold) in both genotypes. Both E2 and T treatments significantly suppressed LHbeta mRNA and serum LH levels in CAST WT males. However, E2 was completely ineffective, and T was only partially effective in suppressing these two indexes in the CAST ERKO males. DHT treatments stimulated a 50% increase in LHbeta mRNA and serum LH levels in WT males, whereas serum LH was significantly suppressed in DHT-treated ERKO males. Although the pituitaries from intact ERKO males contained similar amounts of FSHbeta mRNA, serum FSH levels were 20% higher than those in the intact WT males. Castration increased FSHbeta mRNA levels only in WT males, but significantly increased serum FSH levels in both genotypes. Both E2 and T treatments significantly suppressed serum FSH in CAST WT males, whereas only E2 suppressed FSHbeta mRNA. DHT treatments of CAST WT mice stimulated a small increase in serum FSH, but failed to alter FSHbeta mRNA levels. None of the steroid treatments exerted any significant effect on FSHbeta mRNA or serum FSH levels in CAST ERKOs. These data suggest that hypothalamic GnRH contents can be maintained solely through AR signaling pathways. However, normal regulation of gonadotrope function requires aromatization of T and activation of ERalpha signaling pathways in the gonadotrope. In addition, serum FSH levels in male ERKOs appear to be regulated largely by nonsteroidal testicular factors such as inhibin. Finally, these data suggest that hypothalamic ERbeta may not be involved in mediating the negative feedback effects of T on serum LH and FSH in male mice. PMID- 9751488 TI - Null mutation of the prolactin receptor gene produces a defect in maternal behavior. AB - We have studied pup-directed maternal behavior in mice carrying a germ line null mutation of the PRL receptor (PRLR) gene. Homozygous mutant and heterozygous mutant nulliparous females show a deficiency in pup-induced maternal behavior. Moreover, primiparous heterozygous females exhibit a profound deficit in maternal care when challenged with foster pups. Morris maze studies revealed normal configural learning in the heterozygous and homozygous animals. Eating, locomotor activity, sexual behavior, and exploration (all processes regulated by the hypothalamus) are normal in PRLR mutant mice. Olfactory function was tested in an aversive conditioning paradigm, results indicating that heterozygous and homozygous PRLR mutant mice are not anosmic. These studies clearly establish the PRLR as a regulator of maternal behavior. PMID- 9751490 TI - A high affinity gonadotropin-releasing hormone (GnRH) tracer, radioiodinated at position 6, facilitates analysis of mutant GnRH receptors. AB - Cloning of GnRH receptors from several animal species has made it possible to investigate receptor function using site-directed mutagenesis. However, many mutant GnRH receptors exhibit decreased ligand binding, which makes analysis of their ligand binding characteristics technically difficult. To increase the affinity of binding to the GnRH receptor, a novel tracer ligand, 125I-[His5,D Tyr6]GnRH, was designed and synthesized to allow radioiodination at position 6 rather than the usual position 5. In competition binding assays, total binding of 125I-[His5,D-Tyr6]GnRH was higher than binding of a conventional tracer ligand, 125I-[D-Ala6,N-MeLeu7,Pro9NHEt]GnRH. The bindable fractions and specific activities of both peptides were similar, and the receptor binding affinities of the unlabeled peptides were indistinguishable. However, comparison of the radiolabeled peptides in saturation binding assays showed that the affinity of the peptide, 125I-[His5,D-Tyr6]GnRH, (Kd, 0.19 nM), was approximately 2-fold higher than that of the conventional tracer. The increased binding of 125I [His5,D-Tyr6] GnRH has allowed the development of a sensitive GnRH receptor binding assay for analysis of mutant GnRH receptors that exhibit decreased ligand binding. PMID- 9751489 TI - Regulation of glucagon-like peptide-1 synthesis and secretion in the GLUTag enteroendocrine cell line. AB - Glucagon-like peptide-1 (GLP-1) released from the intestine is a potent stimulator of glucose-dependent insulin secretion. To elucidate the factors regulating GLP-1 secretion, we have studied the enteroendocrine GLUTag cell line. GLP-1 secretion was stimulated in a dose-dependent fashion by activation of protein kinase A or C with forskolin or phorbol 12,13-dibutyrate, respectively (by 2.3 +/- 0.5-fold at 100 microM and 4.3 +/- 0.6-fold at 0.3 microM, respectively; P < 0.01-0.001). Of the regulatory peptides tested, only glucose dependent insulinotropic peptide stimulated the release of GLP-1 (by 2.3 +/- 0.2 fold at 0.1 microM; P < 0.001); glucagon was without effect, and paradoxically, the inhibitory neuropeptide somatostatin-14 increased secretion slightly (by 1.6 +/- 0.3-fold at 0.01 microM; P < 0.05). In tests of several neurotransmitters, only the cholinergic agonists carbachol and bethanechol stimulated peptide secretion in a dose-dependent fashion (by 2.3 +/- 0.5- and 1.7 +/- 0.3-fold at 1000 microM; P < 0.05-0.001); the beta-adrenergic agonist isoproterenol and the chloride channel inhibitor gamma-aminobutyric acid did not affect release of GLP 1. Long chain monounsaturated fatty acids (18:1), but not saturated fatty acids (16:0), also stimulated the release of GLP-1 (by 1.7 +/- 0.1-fold at 150 microM; P < 0.001). Consistent with the presence of a cAMP response element in the proglucagon gene, activation of the protein kinase A-dependent pathway with forskolin increased proglucagon messenger RNA transcript levels by 2-fold (P < 0.05); glucose-dependent insulinotropic peptide and phorbol 12,13-dibutyrate were without effect. Therefore, by comparison with results obtained using primary L cell cultures or in vivo models, GLUTag cells appear to respond appropriately to the regulatory mechanisms controlling intestinal GLP-1 secretion. PMID- 9751491 TI - Effects of insulin-like growth factor administration and bone marrow transplantation on thymopoiesis in aged mice. AB - There has been considerable interest in using hormone replacement therapy to rejuvenate the involuted thymus during aging. GH and insulin-like growth factor-I (IGF-I), a mediator of GH actions, have been of particular interest because of their thymopoietic effects and the fact that their serum concentrations decline during aging. However, treatment of aging rodents with either GH or IGF-I does not restore thymus cellularity to levels present in young animals, suggesting that additional defects might limit the magnitude of their effects. In particular, deficiencies have been reported to accumulate in the bone marrow T cell precursor compartment during aging. In view of this, 18-month-old mice were administered either recombinant IGF-I, bone marrow cells from young mice, or a combination of IGF-I and young bone marrow cells. Thymus cellularity in the latter group of mice was significantly higher than in animals treated with hormone or bone marrow transplantation alone, suggesting that optimal therapies for restoring thymus cellularity must address both endocrine and hematopoietic defects that accumulate during aging. Results from in vitro studies using fetal thymic organ cultures suggest that IGF-I acts by potentiating thymic colonization by bone marrow T cell precursors and/or that the hormone affects some other event soon after thymus colonization. PMID- 9751492 TI - Effects of estrous cyclicity on the expression of the galanin receptor Gal-R1 in the rat preoptic area: a comparison with the male. AB - Variations in the number of galanin receptor (Gal-R1)-expressing cells and levels of Gal-R1 messenger RNA (mRNA) were determined in the preoptic area in intact female rats throughout the phases of the estrous cycle and compared with those in the male. Female and male Wistar rats were fixed by perfusion with 4% paraformaldehyde. Cryostat sections were hybridized with a 35S-labeled antisense Gal-R1 riboprobe. The number of Gal-R1 mRNA-expressing cells was lower in the rostral preoptic area than in the medial preoptic area. During the estrous cycle, the highest number of Gal-R1 mRNA-expressing cells in the rostral preoptic region was detected at 0800 h on proestrus, whereas in the medial preoptic area, the maximum number was observed at 1800 h on estrus. Gal-R1 mRNA levels in individual cells were low during diestrus and increased at estrus in both areas. In the male, the number of mRNA-expressing cells and the hybridization signal were significantly lower than those in females during estrus. The results demonstrate that Gal-R1 gene expression in the preoptic area varies during the estrous cycle and is low in males. Short term treatment of ovariectomized rats with estradiol plus progesterone caused significantly decreased preoptic Gal-R1 mRNA levels compared with those after treatment with estrogen only. These observations suggest that in the preoptic area, expression of Gal-R1 is influenced by progesterone. The variation in Gal-R1 expression is likely to influence the extent to which galanin can influence the preoptic cells implicated in the control of neighboring GnRH cells. PMID- 9751493 TI - Isolation, primary structure, and effects on alpha-melanocyte-stimulating hormone release of frog neurotensin. AB - Neurotensin (NT) was isolated in pure form from the small intestine of the European green frog, Rana ridibunda, and its primary structure was established as pGlu-Ala-His-Ile-Ser-Lys-Ala-Arg-Arg-Pro-Tyr-Ile-Leu. This sequence contains five amino acid substitutions (Leu2-->Ala, Tyr3-->His, Glu4-->Ile, Asn5-->Ser, and Pro7-->Ala) compared with human NT. A peptide with identical chromatographic properties was identified in an extract of frog brain. Synthetic frog NT produced a concentration-dependent increase in alphaMSH release from perifused frog pars intermedia cells, with an ED50 of 5 x 10(-9) M. A maximum response (276.3 +/- 45.5% above basal release) was produced by a 10(-8) M concentration. Repeated administration of NT to melanotrope cells revealed the occurrence of a rapid and pronounced desensitization mechanism. The data are consistent with a possible role for the peptide as a hypophysiotropic factor in amphibians. PMID- 9751494 TI - Alpha2-Heremans Schmid glycoprotein inhibits insulin-stimulated Elk-1 phosphorylation, but not glucose transport, in rat adipose cells. AB - Alpha2-Heremans Schmid glycoprotein (alpha2-HSG) is a member of the fetuin family of serum proteins whose biological functions are not completely understood. There is a consensus that alpha2-HSG plays a role in the regulation of tissue mineralization. However, one aspect of alpha2-HSG function that remains controversial is its ability to inhibit the insulin receptor tyrosine kinase and the biological actions of insulin. Interestingly, some studies suggest that alpha2-HSG differentially inhibits mitogenic, but not metabolic, actions of insulin. However, these previous studies were not carried out in bona fide insulin target cells. Therefore, in the present study we investigate the effects of alpha2-HSG in the physiologically relevant rat adipose cell. We studied insulin-stimulated translocation of the insulin-responsive glucose transporter GLUT4 in transfected rat adipose cells overexpressing human alpha2-HSG. In addition, we measured insulin-stimulated glucose transport in adipose cells cultured with conditioned medium from the transfected cells as well as in freshly isolated adipose cells treated with purified human alpha2-HSG. Compared with control cells, we were unable to demonstrate any significant effect of alpha2-HSG on insulin-stimulated translocation of GLUT4 or glucose transport. In contrast, we did demonstrate that overexpression of alpha2-HSG in adipose cells inhibits both basal and insulin-stimulated phosphorylation of Elk-1 (a transcription factor phosphorylated and activated by mitogen-activated protein kinase and other related upstream kinases). Interestingly, we did not observe any major effects of alpha2-HSG to inhibit insulin-stimulated phosphorylation of the insulin receptor, insulin receptor substrate-1, -2, or -3, in either transfected or freshly isolated adipose cells. We conclude that alpha2-HSG inhibits insulin-stimulated Elk-1 phosphorylation, but not glucose transport, in adipose cells by a mechanism that may involve effector molecules downstream of insulin receptor substrate proteins. PMID- 9751495 TI - A protective role for heme oxygenase expression in pancreatic islets exposed to interleukin-1beta. AB - Heme oxygenase (HO)-1 expression was investigated in rat isolated pancreatic islets. Freshly isolated islets showed no evidence of HO-1 expression. After a 20 h culture, there was a small increase in HO-1 in control islets, and interleukin 1beta (IL-1beta) induced HO-1 expression above control levels. N(G)-monomethyl-L arginine inhibited the IL-1beta-induced increase in HO-1. Sodium nitroprusside generated nitric oxide also increased HO-1 expression. CoCl2 induced a concentration- and time-dependent increase in HO-1, but not heat shock protein 70, expression. Cobalt chloride (CoCl2) protected islets from the inhibitory effects of IL-1beta on glucose-stimulated insulin release and glucose oxidation. Nickel chloride did not mimic the effects of CoCl2. An inhibitor of HO-1 activity, zinc-protoporphyrin IX (ZnPP), prevented the protective effect of CoCl2 on insulin release with IL-1beta but did not affect HO-1 expression or the inhibitory response to IL-1beta alone. ZnPP also inhibited the protective effect of hemin in IL-1beta-treated islets. CoCl2 inhibited the marked increase in islet nitrite production in response to IL-1beta. Cobalt-protoporphyrin IX (CoPP), which increased HO expression and activity, also protected islets from the inhibitory effects of IL-1beta, even though IL-1beta largely blocked the CoPP induced increase in HO-1 expression. In betaHC9 cells, CoCl2 increased HO-1 expression and HO activity, whereas CoPP directly activated HO. ZnPP inhibited basal and CoCl2-stimulated HO activity. Thus, increased HO-1 expression and/or HO activity in response to CoCl2, CoPP, and hemin, seems to mediate protective responses of pancreatic islets against IL-1beta. HO-1 may be protective of beta cells because of the scavenging of free heme, the antioxidant effects of the end product bilirubin, or the generation of carbon monoxide, which might have insulin secretion-promoting effects and inhibitory effects on nitric oxide synthase. PMID- 9751496 TI - Estrogen receptor expression and function in long-term estrogen-deprived human breast cancer cells. AB - Hormone-dependent breast cancer responds to primary therapies that block estrogen production or action, but tumor regrowth often occurs 12-18 months later. Additional hormonal treatments that further reduce estrogen synthesis or more effectively block its action cause additional remissions, but the mechanisms responsible for these secondary responses are not well understood. As a working hypothesis, we postulated that primary hormonal therapy induces adaptive changes, resulting in enhanced estrogen receptor (ER) expression and target gene activation and, further, that secondary treatment modalities interfere with these receptor-mediated transcriptional pathways. To test this hypothesis, we used an MCF-7 breast cancer model system involving deprivation of estradiol in culture for a prolonged period. These long-term estradiol-deprived (LTED) cells adapt by acquiring the ability to regrow in the absence of added estradiol. The experimental paradigm involved the comparison of wild-type cells with LTED cells. As endpoints, we directly assessed ER expression at the messenger RNA-, protein-, and ligand-binding levels and ER functionality by quantitating reporter gene activation and expression of endogenous estrogen target gene messenger RNA, as well as ER coactivator levels. Our data demonstrated an adaptive increase in ER expression and in basal ER functionality, as assessed by read-out of three different transfected reporters in LTED, as opposed to wild-type MCF-7 cells. Increased reporter gene read-out was dramatically inhibited by the pure antiestrogen ICI 182,780. As verification that endogenous (as well as transfected) estrogen target genes had enhanced transcription, we found that the basal levels of c-myb and c-myc message were substantially increased in LTED cells and could be inhibited by antiestrogen. Interestingly, the levels of c-myb and c-myc message in the LTED cells seemed to be increased out of proportion to the degree of ER reporter gene activation and were similar to those in wild-type cells maximally stimulated with estradiol. In addition, not all estrogen responsive genes were activated, because transforming growth factor-alpha message level was not increased in LTED cells. Up-regulation of the steroid receptor coactivator SRC-1 did not seem to mediate the process of enhanced ER-induced transcription. Considering these observations together, we suggest that long-term estradiol deprivation causes adaptive processes that not only involve up regulation of the ER but also influence the specificity and magnitude of activation of estrogen-responsive genes. PMID- 9751497 TI - Systemic challenge with endotoxin stimulates corticotropin-releasing hormone and arginine vasopressin secretion into hypophyseal portal blood: coincidence with gonadotropin-releasing hormone suppression. AB - We tested the hypothesis that systemic immune/inflammatory challenge (endotoxin) activates the neuroendocrine stress axis centrally by stimulating the secretion of CRH and arginine vasopressin (AVP) into hypophyseal portal blood. In addition, we examined the temporal association between this stimulation of the stress neuropeptides and the inhibition of pulsatile GnRH and LH secretion. Using alert, normally behaving ewes, hypophyseal portal and peripheral blood were sampled simultaneously at 10-min intervals for 14 h. Temperature was monitored remotely by telemetry at the same interval. Endotoxin (400 ng/kg, i.v. bolus) or saline as a control was injected after a 4-h baseline period. Portal blood was assayed for CRH, AVP, and GnRH, and peripheral blood was assayed for cortisol, progesterone, and LH. In controls, hypophyseal portal CRH and AVP remained just above or at assay sensitivity, and cortisol showed a regular rhythmic pattern unaffected by saline and typical of basal secretion. In contrast, endotoxin potently stimulated CRH and AVP secretion into portal blood, and cortisol and progesterone into peripheral blood. Both CRH and AVP generally rose and fell simultaneously, although the peak of the AVP response was approximately 10-fold greater than that of CRH. The AVP in portal blood was not due to recirculation of hormone secreted into the peripheral circulation by the posterior pituitary gland, because the AVP increase in peripheral blood was negligible relative to the marked increase in portal blood. The stimulation of CRH and AVP coincided with significant suppression of GnRH and LH pulsatile secretion in these same ewes and with the generation of fever. We conclude that endotoxin induces central activation of the neuroendocrine stress axis, stimulating both CRH and AVP release into the hypophyseal portal blood of conscious, normally behaving ewes. This response is temporally coupled to inhibition of pulsatile GnRH and LH release as well as with stimulation of adrenal cortisol and progesterone secretion and generation of fever. PMID- 9751498 TI - A protease-resistant form of insulin-like growth factor (IGF) binding protein 4 inhibits IGF-1 actions. AB - Smooth muscle cells (SMC) secrete a serine protease that cleaves insulin-like growth factor (IGF) binding protein (IGFBP)-4 into fragments that have low affinity for IGF-1. When IGFBP-4 is added to monolayer cultures of cell types that do not secrete this protease, IGF-1 stimulation of DNA synthesis is significantly inhibited. In contrast, if cell types that secrete this protease are used, IGFBP-4 is a much less potent inhibitor. These studies were conducted to determine whether proteolysis of IGFBP-4 accounted for its reduced capacity to inhibit IGF-1-stimulated DNA synthesis. The cleavage site in IGFBP-4 that the SMC protease uses was determined to be lysine120, histidine121. A protease-resistant mutant form of IGFBP-4 was prepared, expressed, purified, and tested for biologic activity using porcine SMC cultures. Addition of the protease-resistant mutant resulted in inhibition of DNA and cell migration responses to IGF-1. The inhibition was concentration dependent and was maximal when 500 ng/ml (20 nM) of the mutant was added with 20 ng/ml (2.8 nM) of IGF-1. When the mutant was added in the absence of IGF-1, it had no activity. The results show that cleavage of IGFBP-4 at lysine120, histidine121 results in inactivation of the ability of IGFBP-4 to bind to IGF-1. Creation of a mutant form of IGFBP-4 that was not cleaved by the protease resulted in inhibition of IGF-1-stimulated actions. The results suggest that IGFBP-4 can act as a potent inhibitor of the anabolic effects of IGF-1 and that the variables that regulate protease activity may indirectly regulate IGF-1 actions. PMID- 9751499 TI - Sex differences in the daily rhythm of vasoactive intestinal polypeptide but not arginine vasopressin messenger ribonucleic acid in the suprachiasmatic nuclei. AB - The timing of the preovulatory surge of LH in female rodents is tightly coupled to the environmental light/dark cycle. This coupling is mediated by the circadian pacemaker located in the suprachiasmatic nuclei (SCN). Studies indicate that vasoactive intestinal polypeptide (VIP) and arginine vasopressin (AVP), which are synthesized in the SCN, transmit circadian information from the SCN to GnRH neurons, thereby regulating the timing of the LH surge. However, to date, the rhythmic expression of these two peptides in the SCN has only been examined in males. The pattern of VIP expression in males is difficult to reconcile with its role in the LH surge. The purpose of the present study was to assess the rhythm of VIP messenger RNA (mRNA) levels in the SCN of female rats under several endocrine conditions. We compared this rhythm to that in males and to AVP mRNA rhythms in all experimental groups. In all groups of females, VIP mRNA levels were rhythmic, with peak expression occurring during the light phase and a nadir occurring during the dark phase. The rhythm was approximately 12 h out of phase compared with that in males. The rhythmic expression of AVP mRNA in the SCN was virtually identical in all groups of animals. Based on these results, we conclude that 1) the rhythm of VIP seen in the SCN of females during the day may serve as a facilitory signal from the SCN to GnRH neurons; 2) the sex-specific pattern of VIP mRNA does not depend on estradiol; and 3) AVP gene expression within the SCN is not sexually differentiated or altered by estradiol. PMID- 9751500 TI - Lack of coactivator interaction can be a mechanism for dominant negative activity by mutant thyroid hormone receptors. AB - We studied the interactions of two natural thyroid hormone receptor (TR) mutants from patients with resistance to thyroid hormone (RTH) and an artificial TR mutant with a nuclear receptor corepressor, N-CoR, and a steroid receptor coactivator, SRC-1. In electrophoretic mobility shift assays, wild-type TRbeta-1 interacted with N-CoR in the absence of ligand, whereas T3 caused dissociation of the TRbeta-1/N-CoR complex and formation of TRbeta-1/SRC-1 complex. In contrast, a natural mutant (G345R) with poor T3-binding affinity formed TRbeta-1/N-CoR complex, both in the absence and presence of T3, but could not form TRbeta-1/SRC 1 complex. Another TR mutant, which bound T3 with normal affinity and containing a mutation in the AF-2 region (E457D), had normal interactions with N-CoR but could not bind SRC-1. Both these mutants had strong dominant negative activity on wild-type TR transactivation. Studies with a TR mutant that had slightly decreased T3-binding affinity (R320H) showed a T3-dependent decrease in binding to N-CoR and increase in binding to SRC-1 that reflected its decreased ligand binding affinity. Additionally, when N-CoR and SRC-1 were added to these receptors at various T3 concentrations in electrophoretic mobility shift assays, TR/N-CoR and TR/SRC-1 complexes, but not intermediate complexes were observed, suggesting that N-CoR release is necessary before SRC-1 binding to TR. Our data provide new insight on the molecular mechanisms of dominant negative activity in RTH and suggest that the inability of mutant TRs to interact with coactivators such as SRC-1, which results from reduced T3-binding affinity, is a determinant of dominant negative activity. PMID- 9751501 TI - Different functional domains of GLUT2 glucose transporter are required for glucose affinity and substrate specificity. AB - GLUT2 is the major glucose transporter in pancreatic beta-cells and hepatocytes. It plays an important role in insulin secretion from beta-cells and glucose metabolism in hepatocytes. To better understand the molecular determinants for GLUT2's distinctive glucose affinity and its ability to transport fructose, we constructed a series of chimeric GLUT2/GLUT3 proteins and analyzed them in both Xenopus oocytes and mammalian cells. The results showed the following. 1) GLUT3/GLUT2 chimera containing a region from transmembrane segment 9 to part of the COOH-terminus of GLUT2 had Km values for 3-O-methylglucose similar to those of wild-type GLUT2. Further narrowing of the GLUT2 component in the chimeric GLUTs lowered the Km values to those of wild-type GLUT3. 2) GLUT3/GLUT2 chimera containing a region from transmembrane segment 7 to part of the COOH-terminus of GLUT2 retained the ability to transport fructose. Further narrowing of this region in the chimeric GLUTs resulted in a complete loss of the fructose transport ability. 3) Chimeric GLUTs with the NH2-terminal portion of GLUT2 were unable to express glucose transporter proteins in either Xenopus oocytes or mammalian RIN 1046-38 cells. These results indicate that amino acid sequences in transmembrane segments 9-12 are primarily responsible for GLUT2's distinctive glucose affinity, whereas amino acid sequences in transmembrane segments 7-8 enable GLUT2 to transport fructose. In addition, certain region(s) of the amino terminus of GLUT2 impose strict structural requirements on the carboxy-terminus of the glucose transporter protein. Interactions between these regions and the carboxy-terminus of GLUT2 are essential for GLUT2 expression. PMID- 9751502 TI - Alternative splicing of the D2 dopamine receptor messenger ribonucleic acid is modulated by activated sex steroid receptors in the MMQ prolactin cell line. AB - The two isoforms of the D2 dopamine receptor are generated by alternative splicing of the exon 6 of the premessenger RNA (pre-mRNA), changing the length of the third cytoplasmic loop involved in the coupling to G proteins. In the MMQ PRL cell line, sex steroid hormones modulated the proportion of the two D2 receptor isoforms. Under controlled culture conditions, 17beta-estradiol (E2) strongly favored the production of the long isoform of D2 mRNA over the short one, whereas both isoforms were equally abundant when culture medium was hormone depleted. In the presence of progesterone (P), E2 action was inhibited, and equal amounts of each D2 receptor isoform were produced in the cells. Hormone treatments never modified either the total amount of D2 receptor mRNA and D2 receptor binding sites or D2 receptor-mediated inhibition of adenylyl cyclase. Specific antagonists demonstrated that the activity of each hormone depended on their nuclear receptors. Inhibitors of gene transcription or translation also showed that their activity required protein synthesis. The expression of the short D2 receptor isoform was never prominent, even at the single cell level. Analysis of the intron sequence flanking alternative exon 6 showed that only the upstream intron presented two sequence tracts known to be targets for splicing factors. Taken together, these results provide converging evidence for a physiologically relevant mechanism by which sex steroid receptors could regulate the expression of a splicing factor favoring the production of the long dopamine D2 receptor isoform. PMID- 9751503 TI - Galanin is a paracrine inhibitor of gonadotroph function in the female rat. AB - Recent evidence suggests that pituitary galanin synthesized in the lactotroph is a paracrine regulator of lactotroph proliferation and PRL secretion and that these effects are mediated via a pituitary-specific galanin receptor, GAL R2(orig.). At this receptor subtype, the galanin fragment 3-29 is fully active, in contrast to both the cloned GAL-R1 and GAL-R2, at which this fragment is inactive. Since paracrine communication has been demonstrated between pituitary gonadotrophs and lactotrophs, we investigated the hypothesis that galanin is also a paracrine regulator of gonadotroph function. Galanin attenuated LHRH-stimulated LH release in a dose-dependent manner in monodispersed rat anterior pituitaries harvested at proestrus (LHRH 100 nM, 10.7 +/- 0.2 ng/ml(-1) x 4 h vs. LHRH 100 nM + 1 microM porcine galanin (pGal), 7.0 +/- 0.2 ng/ml(-1) x 4 h; P < 0.01; i.e. 37% reduction). Galanin had similar suppressive effects on FSH release. Galanin, also dose-dependently, attenuated the LHRH-stimulated LH release from perifused proestrous rat pituitary fragments. pGal (1 microM) reduced the stimulated LH release by 80%, [area under the curve (AUC), LHRH 100 nM, 713 +/- 149 vs. LHRH 100 nM + 1 microM pGal, 131 +/- 7 ng/min x ml(-1) x 4 h; P < 0.02]. In addition, galanin 3-29, the specific GAL-R2(orig.) receptor agonist, inhibited LHRH stimulated LH release from perifused proestrous rat pituitary fragments [AUC, LHRH 100 nM, 642 +/- 77 ng/min x ml(-1) vs. LHRH 100 nM + pGal 1-29, 206 +/- 44 ng/min x ml(-1) (P < 0.02); and LHRH 100 nM + pGal 3-29, 310 +/- 19 ng/min x ml( 1) (P < 0.02)]. Immunoblockade with specific galanin antiserum potentiated the LHRH-stimulated release of LH by 48% from perifused proestrous rat pituitary fragments (AUC, LHRH 100 nM + galanin antiserum, 721 +/- 65 ng/min x ml(-1) vs. LHRH 100 nM alone or with nonimmune antiserum, 489 +/- 33 ng/min x ml(-1) or 545 +/- 46 ng/min x ml(-1), P < 0.05). This data suggests that galanin may act as a paracrine agent via the pituitary-specific GAL-R2(orig.) to inhibit gonadotroph function. PMID- 9751504 TI - Developmental changes in galanin receptors in the quail oviduct and the effect of ovarian sex steroids on galanin receptor induction. AB - We have recently isolated an oviposition-inducing peptide from mature quail oviducts identified as avian galanin. This peptide evoked vigorous contractions of the uterine oviduct through binding to receptors located in the uterus. The questions arising from these findings are: what changes occur in galanin receptors in the uterus during maturation, and what is the hormonal factor(s) that induces uterine galanin receptors? Therefore, the present study examined changes in uterine galanin receptors with age and the effect of administration of ovarian sex steroids on galanin receptors in the quail. Immature females reared under long day (LD) photoperiods from 4 weeks of age demonstrated a progressive increase in specific galanin binding per both unit uterine weight and per whole uterus concurrent with uterine development during 4-13 weeks. Scatchard plot analyses of the binding to the uterine preparation showed that the equilibrium dissociation constant (Kd) was about 0.30-0.34 nM regardless of age, and the change in galanin binding during uterine development was due to a change in the number of binding sites. Plasma 17beta-estradiol and progesterone concentrations were almost constant between 4-6 weeks and tended to increase thereafter. Administration of 17beta-estradiol to immature females for 1 week increased not only uterine weight but also specific galanin binding per unit uterine weight, whereas progesterone increased only the binding per unit uterine weight. Both sex steroids also induced an increase in total binding per uterus. Combined administration of 17beta-estradiol and progesterone induced marked increases in the galanin binding, and the effect was not additive but, rather, was synergistic. Scatchard plot analysis showed that the number of binding sites, but not the Kd, was increased by steroid treatment. Administration of 17beta estradiol or progesterone increased each circulating steroid level to that relatively similar to the maximal levels observed in females exposed to LD. Thus, ovarian sex steroids may contribute at least in part as hormonal factors to galanin receptor induction, which takes place in the uterine oviduct during development. PMID- 9751505 TI - DAX-1 blocks steroid production at multiple levels. AB - DAX-1 is an unusual member of the nuclear hormone receptor superfamily whose expression is mainly, but not uniquely, restricted to steroidogenic tissues. We have recently shown that DAX-1 can block the first and rate-limiting step in steroid biosynthesis by repressing StAR (steroidogenic acute regulatory protein) expression. Here we show that DAX-1 blocks steroid production at multiple levels in the Y-1 mouse adrenocortical tumor cell line. Expression of DAX-1 in Y-1 cells significantly impairs both basal and cAMP-stimulated steroid production, without affecting the functionality of the cAMP-responsive PKA pathway. Experiments using an hydroxylated cholesterol derivative show that biochemical steps in steroidogenesis subsequent to cholesterol delivery to mitochondria are also impaired in Y-1 cells expressing DAX-1. This is explained by the repression of P450scc and 3beta-HSD expression, in addition to StAR. DAX-1 expression in Y-1 cells results in the inhibition of the activity of the StAR, P450scc and 3beta HSD promoters. An inappropriate steroidogenic block in the male fetus might have an important role in the pathogenesis of sex reversal syndromes caused by a duplication of the genomic region of the X chromosome containing the DAX-1 gene. PMID- 9751506 TI - Effect of prenatal exposure to diethylstilbestrol on Mullerian duct development in fetal male mice. AB - The clinical use of diethylstilbestrol (DES) by pregnant women has resulted in an increased incidence of genital carcinoma in the daughters born from these pregnancies. Also, in the so-called DES-sons abnormalities were found, mainly, the presence of Mullerian duct remnants, which indicates that fetal exposure to DES may have an effect on male sex differentiation. Fetal regression of the Mullerian ducts is under testicular control through anti-Mullerian hormone (AMH). In male mice, treated in utero with DES, the Mullerian ducts do not regress completely, although DES-exposed testes do produce AMH. We hypothesized that incomplete regression in DES-exposed males is caused by a diminished sensitivity of the Mullerian ducts to AMH. Therefore, the effect of DES on temporal aspects of Mullerian duct regression and AMH type II receptor (AMHRII) messenger RNA (mRNA) expression in male mouse fetuses was studied. It was observed that Mullerian duct regression was incomplete at E19 (19 days post coitum), upon DES administration during pregnancy from E9 through E16. Furthermore, analysis of earlier time points of fetal development revealed that the DES treatment had clearly delayed the onset of Mullerian duct formation by approximately 2 days; in untreated fetuses, Mullerian duct formation was complete by E13, whereas fully formed Mullerian ducts were not observed in DES-treated male fetuses until E15. Using in situ hybridization, no change in the localization of AMH and AMHRII mRNA expression was observed in DES-exposed male fetuses. The mRNA expression was quantified using ribonuclease protection assay, showing an increased expression level of AMH and AMHRII mRNAs at E 13 in DES-exposed male fetuses. Furthermore, the mRNA expression levels of Hoxa 11 and steroidogenic factor-1 (SF-1) were determined as a marker for fetal development. Prenatal DES exposure had no effect on Hoxa 11 mRNA expression, indicating that DES did not exert an overall effect on the rate of fetal development. In DES-exposed male fetuses, SF-1 showed a similar increase in mRNA expression as AMH, in agreement with the observations that the AMH gene promoter requires an intact SF-1 DNA binding site for time- and cell-specific expression, although an effect of DES on SF-1 expression in other tissues, such as the adrenal and pituitary gland, cannot be excluded. However, the increased expression levels of AMH and AMHRII mRNAs do not directly explain the decreased sensitivity of the Mullerian ducts to AMH. Therefore, it is concluded that prenatal DES exposure of male mice delays the onset of Mullerian duct development, which may result in an asynchrony in the timing of Mullerian duct formation, with respect to the critical period of Mullerian duct regression, leading to persistence of Mullerian duct remnants in male mice. PMID- 9751507 TI - Interaction of estrogenic chemicals and phytoestrogens with estrogen receptor beta. AB - The rat, mouse and human estrogen receptor (ER) exists as two subtypes, ER alpha and ER beta, which differ in the C-terminal ligand-binding domain and in the N terminal transactivation domain. In this study, we investigated the estrogenic activity of environmental chemicals and phytoestrogens in competition binding assays with ER alpha or ER beta protein, and in a transient gene expression assay using cells in which an acute estrogenic response is created by cotransfecting cultures with recombinant human ER alpha or ER beta complementary DNA (cDNA) in the presence of an estrogen-dependent reporter plasmid. Saturation ligand-binding analysis of human ER alpha and ER beta protein revealed a single binding component for [3H]-17beta-estradiol (E2) with high affinity [dissociation constant (Kd) = 0.05 - 0.1 nM]. All environmental estrogenic chemicals [polychlorinated hydroxybiphenyls, dichlorodiphenyltrichloroethane (DDT) and derivatives, alkylphenols, bisphenol A, methoxychlor and chlordecone] compete with E2 for binding to both ER subtypes with a similar preference and degree. In most instances the relative binding affinities (RBA) are at least 1000-fold lower than that of E2. Some phytoestrogens such as coumestrol, genistein, apigenin, naringenin, and kaempferol compete stronger with E2 for binding to ER beta than to ER alpha. Estrogenic chemicals, as for instance nonylphenol, bisphenol A, o, p'-DDT and 2',4',6'-trichloro-4-biphenylol stimulate the transcriptional activity of ER alpha and ER beta at concentrations of 100-1000 nM. Phytoestrogens, including genistein, coumestrol and zearalenone stimulate the transcriptional activity of both ER subtypes at concentrations of 1-10 nM. The ranking of the estrogenic potency of phytoestrogens for both ER subtypes in the transactivation assay is different; that is, E2 >> zearalenone = coumestrol > genistein > daidzein > apigenin = phloretin > biochanin A = kaempferol = naringenin > formononetin = ipriflavone = quercetin = chrysin for ER alpha and E2 >> genistein = coumestrol > zearalenone > daidzein > biochanin A = apigenin = kaempferol = naringenin > phloretin = quercetin = ipriflavone = formononetin = chrysin for ER beta. Antiestrogenic activity of the phytoestrogens could not be detected, except for zearalenone which is a full agonist for ER alpha and a mixed agonist antagonist for ER beta. In summary, while the estrogenic potency of industrial derived estrogenic chemicals is very limited, the estrogenic potency of phytoestrogens is significant, especially for ER beta, and they may trigger many of the biological responses that are evoked by the physiological estrogens. PMID- 9751508 TI - Leptin inhibits directly glucocorticoid secretion by normal human and rat adrenal gland. AB - Different interactions have been described between glucocorticoids and the product of the ob gene leptin. Leptin can inhibit the activation of the hypothalamo-pituitary-adrenal axis by stressful stimuli, whereas adrenal glucocorticoids stimulate leptin production by the adipocyte. The present study was designed to investigate the potential direct effects of leptin to modulate glucocorticoid production by the adrenal. Human adrenal glands from kidney transplant donors were dissociated, and isolated primary cells were studied in vitro. These cells were preincubated with recombinant leptin (10(-10)-10(-7) M) for 6 or 24 h, and basal or ACTH-stimulated cortisol secretion was subsequently measured. Basal cortisol secretion was unaffected by leptin, but a significant and dose-dependent inhibition of ACTH-stimulated cortisol secretion was observed [down by 29 +/- 0.1% of controls with the highest leptin dose, P < 0.01 vs. CT (unrelated positive control)]. This effect of leptin was also observed in rat primary adrenocortical cells, where leptin inhibited stimulated corticosterone secretion in a dose-dependent manner (down by 46 +/- 0.1% of controls with the highest leptin dose, P < 0.001 vs. CT). These effects of leptin in adrenal cells are likely mediated by the long isoform of the leptin receptor (OB-R), because its transcript was found to be expressed in the adrenal tissue and leptin had no inhibitory effect in adrenal glands obtained from db/db mice. Therefore, leptin inhibits directly stimulated cortisol secretion from human and rat adrenal glands, and this may represent an important mechanism to modulate glucocorticoid levels in various metabolic states. PMID- 9751509 TI - The effect of 9-cis-retinoic acid on proliferation and differentiation of a spermatogonia and retinoid receptor gene expression in the vitamin A-deficient mouse testis. AB - Retinoid X receptors (RXRs) are key regulators in retinoid signaling. Knowledge about the effects of 9-cis-retinoic acid (9-cis-RA), the natural ligand for the RXRs, may also provide insight in the functions of RXRs. In this study, the effect of 9-cis-RA on spermatogenesis in vitamin A-deficient (VAD) mice was examined. Administration of 9-cis-RA stimulated the differentiation and subsequent proliferation of the growth-arrested A spermatogonia in the testis of VAD mice. However, compared with all-trans-retinoic acid (ATRA), relatively higher doses of 9-cis-RA were necessary. This could not simply be due to a lower or delayed activity of 9-cis-RA, as simultaneous administration of ATRA and 9-cis RA did not cause a synergistic effect. Instead, the presence of 9-cis-RA diminished the effect of ATRA by approximately one third. Studies of in vivo transport and metabolism showed that ATRA and 9-cis-RA, after administration to VAD mice, penetrated the testis equally well. However, 9-cis-RA was metabolized much faster than ATRA, and other metabolites were formed. This may account for the above-described differential effects of ATRA and 9-cis-RA on spermatogenesis. Similar to ATRA, 9-cis-RA transiently induced the messenger RNA expression of the nuclear RA receptor RAR beta, suggesting a role for this receptor in the effects of retinoids on the differentiation and proliferation of A spermatogonia. In contrast, the messenger RNA expression of the nuclear retinoid receptors RXR alpha, -beta, and -gamma was not changed significantly by administration of their ligand, 9-cis-RA. Hence, 9-cis-RA does not seem to exert its effect on spermatogenesis through altered expression of the RXRs. PMID- 9751510 TI - Human thyroperoxidase is largely retained and rapidly degraded in the endoplasmic reticulum. Its N-glycans are required for folding and intracellular trafficking. AB - Human thyroperoxidase (hTPO), a type I transmembrane heme containing glycoprotein, catalyzes iodide organification and thyroid hormone synthesis and plays a major role in thyroid autoimmunity. Whereas hormonosynthesis occurs at the apical membrane of thyroid cells, TPO localizes mainly in the perinuclear membrane and the endoplasmic reticulum. To establish the intracellular trafficking and the structural characteristics of hTPO in the various cell compartments, hTPO was stably expressed in the Chinese hamster ovary cell line, and its folding was studied with two monoclonal antibodies (mAbs): mAb 47, recognizing a linear epitope; and mAb 15, recognizing a conformational epitope present in the mature protein. The results show that only 15-20% of hTPO molecules were able to acquire a conformation suitable for the recognition by mAb 15. On the other hand, only a part (approximately 15%) of the latter were able to reach the plasma membrane. The hTPO, unable to fold correctly, was more rapidly degraded than that recognized by mAb 15 (half-time, 2 h vs. 7 h). Study of the carbohydrate content of hTPO showed that N-glycans with complex-type structure were found only on hTPO at the cell surface, whereas intracellular hTPO bore high mannose-type structures. Taken together, these data demonstrate that the intracellular pool of enzyme is formed of newly synthesized molecules and is not caused by recycling of mature hTPO from the cell surface. Complete inhibition of hTPO N-glycosylation with tunicamycin led to a 95% decrease in hTPO at the plasma membrane and, thus, to a decrease in enzymatic activity at the cell surface, emphasizing the role of N-glycans in the intracellular trafficking of hTPO. However, inhibition of formation of complex-type structures with deoxymannojirimycin and of O-glycans with phenyl-alpha-GalNAc did not influence the intracellular trafficking and enzymatic activity of hTPO. PMID- 9751511 TI - Developmental changes in serum levels of free and total insulin-like growth factor I (IGF-I), IGF-binding protein-1 and -3, and the acid-labile subunit in rats. AB - We have recently described a competitive binding assay for rat insulin-like growth factor-binding protein-3 (IGFBP-3) based on the ability of IGFBP-3 to form a ternary complex with the acid-labile subunit (ALS) in the presence of IGF-I. Using this assay we studied groups of male (n = 6) and female rats (n = 6) at 20, 30, 40, 50, 60, 80, and 130 days of age. Nonfasting serum levels of IGFBP-3 were compared with those of total (extractable) IGF-I (tIGF-I) and ALS as well as IGFBP-3 determined by ligand blotting. Additionally, we studied the relationship between ultrafiltered free IGF-I (fIGF-I) and immunoassayable IGFBP-1. IGFBP-3 was dependent on age only (P < 0.0001), but tended to be higher in males than in females (P = 0.06); between 20-130 days levels increased from 6.5 +/- 1.7 to 73.6 +/- 7.2 nmol/liter in males and from 5.4 +/- 1.6 to 51.3 +/- 8.0 nmol/liter in females. IGFBP-3 correlated positively with tIGF-I (r = 0.90; P < 0.0001), ALS (r = 0.92; P < 0.0001), and IGFBP-3, as determined by ligand blotting (r = 0.88; P < 0.0001). The molar ratio of IGFBP-3 to tIGF-I increased from 0.23 +/- 0.04 to 0.76 +/- 0.04 (P < 0.0001) without any sex dependence. An age- and sex-dependent decrease in IGFBP-1 was observed (P < 0.0001), from 10.9 +/- 2.5 to 1.2 +/- 0.2 nmol/liter in females and from 8.9 +/- 0.7 to 0.2 +/- 0.04 nmol/liter in males. Free IGF-I (fIGF-I) increased with age (from 0.7 +/- 0.2 to 7.1 +/- 0.5 nmol/liter; P < 0.0001), and levels were inversely correlated with IGFBP-1 (r = 0.80; P < 0.0001). In young rats, IGFBP-1 circulated in a 10-fold molar excess over the level of fIGF-I, whereas in older rats, fIGF-I exceeded IGFBP-1 by an average of 9-fold in females and by up to almost 60-fold in males. We conclude that in rats 1) IGFBP-3 and fIGF-I are strongly age dependent; 2) IGFBP-3 correlates positively with ALS and tIGF-I; and 3) fIGF-I and IGFBP-1 are inversely correlated. This is in accordance with clinical findings. However, in humans the adult level of fIGF-I rarely exceeds 0.3 nmol/liter, and IGFBP-1 usually circulates in excess of fIGF-I. Thus, our results also imply species differences in the IGF systems of humans and rats. PMID- 9751512 TI - Type-1 parathyroid hormone (PTH)/PTH-related peptide (PTHrP) receptors activate phospholipase C in response to carboxyl-truncated analogs of PTH(1-34). AB - The carboxyl(C)-truncated human (h) PTH (hPTH) analog hPTH(1-31), which activates adenylyl cyclase (AC), but not protein kinase C, in rat osteosarcoma cells, exerts an anabolic effect on rat bone in vivo similar to that of hPTH(1-34). It has been proposed, therefore, that this action of PTH(1-34) is mediated exclusively by stimulation of AC via the rat type-1 PTH/PTH-related peptide (PTHrP) receptor (PTH1R). To determine whether this selective signaling pattern also might be a property of the hPTH1R, we studied signal transduction via heterologously expressed hPTH1Rs in response to activation by hPTH(1-34), hPTH(1 31), and a C-truncated analog that does not increase rat bone mass in vivo, hPTH(1-30). In porcine LLC-PK1 cells that stably expressed recombinant hPTH1Rs, these three peptides activated AC identically (EC50 = 1-2 nM). In cells with comparable expression of rat PTH1Rs, AC activation by hPTH(1-34) and hPTH(1-31) again was identical, whereas full activation by hPTH(1-30) required higher concentrations (EC50 = 10 nM vs. 1 nM). Surprisingly, hPTH(1-31) fully stimulated phospholipase C (PLC), via both species of PTH1Rs, with potency that was similar (hPTH1Rs) or slightly reduced (rat PTH1Rs), relative to that of hPTH(1-34). hPTH(1-30), however, was 5-fold less potent than hPTH(1-34) in activating PLC via hPTH1Rs and showed weak and only partial activity via the rat PTH1R. Comparable results were obtained when human and rat PTH1Rs were transiently expressed heterologously in COS-7 cells or homologously in HEK 293 and UMR 106-01 cells, respectively. Binding affinities of these C-truncated peptides to human and rat PTH1Rs were concordant with their relative potencies in activating PLC. We conclude that hPTH(1-31) and, to a lesser extent, hPTH(1-30) can activate PLC, as well as AC, via both rat and human PTH1Rs. Accordingly, a role for PLC activation in the anabolic action of PTH in vivo cannot be excluded. PMID- 9751513 TI - Distribution, characterization, and growth hormone-releasing activity of pituitary adenylate cyclase-activating polypeptide in the European eel, Anguilla anguilla. AB - The complementary DNA encoding pituitary adenylate cyclase-activating polypeptide (PACAP) has been cloned from two species of teleost fishes, the Sockeye salmon and the Thai catfish, and the amino acid sequence of PACAP has been determined in another teleost, the stargazer. However, to date, the detailed distribution of PACAP immunoreactivity has never been investigated in the fish brain. In the present study, we have determined the localization of PACAP-immunoreactive neurons in the central nervous system of a primitive teleost fish, the European eel Anguilla anguilla, using an antiserum raised against PACAP27. PACAP-positive perikarya were exclusively observed in the diencephalon, i.e. in the preoptic nucleus of the hypothalamus and in the dorsal and ventral nuclei of the thalamus. PACAP-immunoreactive fibers were detected in various areas of the brain, notably in the ventral telencephalon, the diencephalon, the mesencephalon, the cerebellar valvula, and the medulla oblongata. In addition, a dense accumulation of PACAP containing nerve terminals was found in the pars distalis of the pituitary. The PACAP-like immunoreactivity contained in the eel brain was characterized by HPLC analysis combined with RIA quantification. The major form of PACAP-immunoreactive material coeluted with mammalian PACAP38. Molecular cloning of the PACAP precursor has previously shown that in fish, PACAP and GH-releasing hormone (GHRH) originate from the same precursor. We have thus investigated the effects of PACAP and GHRH on GH secretion from eel pituitary cells in primary culture. Dose-response experiments revealed that PACAP27 and PACAP38 possessed the same efficacy, but PACAP38 was 12 times more potent than PACAP27 in stimulating GH release (ED50 = 4.3 x 10(-10) and 3.5 x 10(-9) M, respectively). In contrast, GHRH, even at a high concentration (10(-6) M), had no effect on GH release. Taken together, these data indicate that in the eel, PACAP may play a significant role in the regulation of somatotrope cells: 1) PACAP-immunoreactive neurons are exclusively located in the diencephalon and send numerous projections in the pars distalis; and 2) PACAP, but not GHRH, dose dependently stimulates GH secretion from cultured eel pituitary cells. PMID- 9751514 TI - Thyroid hormone efflux from monolayer cultures of human fibroblasts and hepatocytes. Effect of lipoproteins and other thyroxine transport proteins. AB - We have previously shown that human skin fibroblasts exposed to preformed low density lipoprotein (LDL)-thyroxine (T4) complexes internalize more T4 than they do when exposed to T4 alone. The system is set to function when the LDL receptor is up-regulated by reducing the intracellular concentration of cholesterol, and the LDL concentration outside the cell is in the range of the kDa of the receptor. High density lipoproteins (HDL), albumin (HSA), transthyretin (TTR), and thyroxine-binding globulin (TBG) interfere with, rather than facilitate, T4 entry. Of the three classes of lipoproteins (VLDL, LDL, and HDL), HDL is the major carrier of thyroid hormones. While LDL delivers cholesterol (and T4) to cells, HDL is the scavenger of cholesterol. We thus hypothesized that HDL could also facilitate thyroid hormone exit from cells. This hypothesis was tested on two human cell lines: skin fibroblasts and hepatocytes (Hep G2), using physiological concentrations of HDL or, as control, physiological concentrations of LDL, HSA, TTR, and TBG or buffer. Because cell surface receptors for HDL are regulated by intracellular cholesterol in a manner opposite to that of LDL receptors, we evaluated the effect of HDL (and other proteins) in three states: normal, high, and low intracellular cholesterol content (i.e. normal, high, and low expression of HDL receptors). In both cell lines and with either T4 or T3, we found that: 1) HDL as well as the other proteins tested increased the efflux and augmented both the initial rate of exit and the equilibrium value. 2) The efflux did not saturate over a wide range of protein concentrations. 3) The effect of HDL, LDL, and the other proteins on the fractional efflux rate of thyroid hormones remained the same irrespective of the intracellular cholesterol content (and, therefore, irrespective of the expression of either LDL or HDL receptors). 4) HSA, TTR, and TBG were, on a mass basis, equipotent and more efficient than lipoproteins. However, the effect of lipoproteins--whose Ka for T4 is comparable to that of HSA--was disproportionately high. On a molar basis, LDL (about 80% of the weight being accounted for by lipids) was more effective than HDL2 (about 60% lipids) and HDL2 was more effective than HDL3 (about 40% lipids), suggesting that the disproportionate effect of lipoproteins was due to transfer of the lypophylic thyroid hormones to the lipid moiety of lipoproteins. 5. A mixture of HDL and LDL gave the same efflux rate as a mixture of HSA, TTR, and TBG. The data indicate that the efflux of T4 and T3 from cells is rapid and appears not to be mediated by a particular lipoprotein. The disproportionately large effect of lipoproteins, which are low affinity thyroid hormone carriers, compared with nonlipoprotein carriers, and the greater effect of LDL compared with HDL, might indicate that the lipoproteins establish a nonspecific physical contact with the plasma membrane and that their hydrophobic nature favors the release of the similarly hydrophobic thyroid hormones. PMID- 9751515 TI - Therapeutic efficacy of 1alpha,25-dihydroxyvitamin D3 and calcium in osteopenic ovariectomized rats: evidence for a direct anabolic effect of 1alpha,25 dihydroxyvitamin D3 on bone. AB - It is an important question for clinical therapy of osteoporosis with vitamin D metabolites whether these compounds exert their beneficial effects on the skeleton indirectly through an increase in intestinal calcium absorption or whether there is also a major direct component of action on bone. In this study, female 6-month-old Fischer rats were either ovariectomized (OVX) or sham operated. One month before surgery, all rats were placed on a diet containing 0.25% calcium and were kept on this diet throughout the study. Beginning 3 months post-OVX, groups of OVX rats orally received vehicle, a calcium supplement, low dose (0.025 microg/kg x day) or high dose (0.1 microg/kg x day) 1alpha,25 dihydroxyvitamin D3 [1,25-(OH)2D3], or combinations of low and high dose 1,25 (OH)2D3 with the calcium supplement. By 3 months postsurgery, pretreatment OVX controls had lost 74% and 37% of tibial and vertebral cancellous bone, respectively. Two-way factorial ANOVA showed that a 3-month treatment of osteopenic OVX rats with 1,25-(OH)2D3 dose dependently increased vertebral and tibial cancellous bone mass (P < 0.001 and P = 0.021, respectively) and trabecular width (P < 0.001). Furthermore, 1,25-(OH)2D3 increased serum calcium (P = 0.028) and urinary calcium excretion (P < 0.001) and reduced serum PTH levels (P < 0.001), osteoclast numbers (P < 0.001), and urinary collagen cross links excretion (P < 0.001). Calcium supplementation alone was without therapeutic effect, and there was no significant two-way interaction between the individual treatment effects of 1,25-(OH)2D3 and calcium on bone mass. These data indicate that the anabolic effects of 1,25-(OH)2D3 in osteopenic OVX rats are mediated through a direct activity on bone. PMID- 9751516 TI - Glucose stimulation of pancreatic beta-cell lines induces expression and secretion of dynorphin. AB - To investigate adaptive responses of pancreatic beta-cells to hyperglycemia, genes induced by glucose stimulation were identified by subtraction cloning. Among 53 clones representing differentially expressed genes, 20 encoded the endogenous opioid precursor, prodynorphin. The amino acid sequence of murine prodynorphin is identical to the rat protein in sequences comprising the opioid peptides and 86% identical in the remainder of the molecule. Stimulation of MIN6 cells increased prodynorphin RNA levels to more than 20-fold in proportion to physiological glucose concentrations. Similar induction levels were observed in murine betaTC3 and rat Rinm5F beta-cell lines. Prodynorphin RNA expression increased within 1 h of glucose stimulation, achieved maximal levels by 4 h, and remained elevated for at least 24 h. By using RIA, MIN6 cells were shown to contain and secrete increased amounts of dynorphin-A following glucose stimulation. Treatment of MIN6 cells with KCl, forskolin, or isobutyl-methyl xanthine strongly induced prodynorphin RNA expression, suggesting that induction may be related to secretion-coupled signaling pathways. The induction of prodynorphin in several beta-cell lines is consistent with previous demonstrations of beta-cell synthesis of other endogenous opioids, including beta endorphin, and suggests that opioids may have a potentially significant role in regulating beta-cell secretion. PMID- 9751517 TI - Calreticulin: an intracellular Ca++-binding protein abundantly expressed and regulated by androgen in prostatic epithelial cells. AB - Calreticulin was identified in a screen for androgen-response genes in the rat ventral prostate. Northern blot and Western blot analyses in the rat model showed that both calreticulin messenger RNA and protein are down-regulated by castration and up-regulated by androgen replacement in the prostate. Northern blot analysis showed that calreticulin expression level in the prostate is much higher than that in seminal vesicles, heart, brain, muscle, kidney, and liver. The regulation of calreticulin expression by androgen is only observed in the prostate and seminal vesicles, two male secondary sex organs. The induction of calreticulin by androgen in prostate organ culture partially resists protein synthesis inhibition, suggesting that calreticulin is a direct androgen-response gene. In situ hybridization and immunohistochemistry studies showed that calreticulin is an intracellular protein in prostatic epithelial cells. Because calreticulin is a major intracellular Ca++-binding protein with 1 high-affinity and 25 low-affinity Ca binding sites, our observations suggest that calreticulin is a promising candidate that mediates androgen regulation of intracellular Ca++ levels and/or signals in prostatic epithelial cells. The expression of calreticulin is also regulated by androgen in the mouse and human prostate, suggesting that androgen regulation and function of calreticulin in the prostate are conserved evolutionarily. PMID- 9751518 TI - Role of stromal and epithelial estrogen receptors in vaginal epithelial proliferation, stratification, and cornification. AB - Estradiol 17-beta (E2) induces epithelial proliferation, stratification, and cornification in vaginal epithelium. Our aim was to determine the respective roles of epithelial and stromal estrogen receptor-alpha (ER alpha) in these E2 induced events. Vaginal epithelium (E) and stroma (S) from adult ER alpha knockout (ko) and wild-type (wt) neonatal Balb/c mice were enzymatically separated and used to produce four types of tissue recombinants in which epithelium, stroma, or both lack functional ER alpha. Tissue recombinants were grafted into female nude mice, which were subsequently ovariectomized and treated with oil or E2. In response to E2 treatment, grafts prepared with wt-S (wt-S + wt E and wt-S + ko-E) showed similar large increases in epithelial labeling index, indicating that E2 stimulated epithelial proliferation despite a lack of epithelial ER alpha in wt-S + ko-E tissue recombinants. Conversely, in tissue recombinants prepared with ko-S (ko-S + wt-E and ko-S + ko-E), epithelial labeling index remained at baseline levels after E2 or oil treatment, even though epithelial ER alpha were detected in ko-S + wt-E grafts. Epithelial cornification was present in wt-S + wt-E grafts from E2-treated hosts, whereas epithelium in all other tissue recombinants failed to cornify. Grafts composed of wt-S + wt-E from E2-treated hosts had highly stratified epithelium, whereas epithelial thickness was reduced almost 60% in wt-S + ko-E tissue recombinants grown in E2 treated hosts and was atrophic in all other tissue recombinants. In addition, cytokeratin 10, a marker of epithelial differentiation, was strongly expressed in wt-S + wt-E tissue recombinants grown in E2-treated hosts but was markedly reduced or absent in all other tissue recombinants. These results indicate that E2-induced vaginal epithelial proliferation is mediated indirectly through stromal ER alpha, consistent with our recent findings in uterus. Conversely, both epithelial and stromal ER alpha are required for E2-induced cornification and normal epithelial stratification. These are the first known functions attributed to epithelial ER alpha in vivo and the first time any epithelial response to E2 has been shown to involve both stromal and epithelial ER alpha. PMID- 9751519 TI - Human osteoclasts and osteoclast-like cells synthesize and release high basal and inflammatory stimulated levels of the potent chemokine interleukin-8. AB - Chemokines, including interleukin-8 (IL-8), function as key mediators in diverse inflammatory disorders via promoting the recruitment, proliferation, and activation of vascular and immune cells. IL-8 levels are elevated in inflammatory diseases, such as rheumatoid arthritis, osteoarthritis, osteomyelitis, and periodontal disease, that also exhibit progressive bone loss. Therefore, it is possible that IL-8 contributes to the osteopenia associated with these pathological conditions. Although macrophages, neutrophils, and endothelial cells are considered the primary sources of inflammation-induced IL-8 increases, we report here for the first time that human bone marrow-derived osteoclast-like cells (hOCL) as well as authentic bone-resorbing human osteoclasts (hOC) isolated from osteoporotic femoral heads express messenger RNA (mRNA) for IL-8 and secrete high levels of IL-8 during culture. Basal IL-8 release by cultured hOC or hOCL was orders of magnitude greater than the release of the proinflammatory cytokines IL-1beta, IL-6, and tumor necrosis factor-alpha. At a cellular level, in situ hybridization analysis revealed that IL-8 mRNA was expressed in resorbing hOC of rheumatoid arthritic pannus and was substantially greater than that expressed in hOC of noninflammatory giant cell tumor of bone tissue. Therefore, the potential inflammation-mediated induction of IL-8 was directly assessed using cultured hOCL. IL-8 release was stimulated by proinflammatory signals (IL-1alpha, tumor necrosis factor-alpha, lipopolysaccharide, or phorbol 12-myristate 13-acetate), unaffected by various other osteotropic modulators (transforming growth factor beta1 and -beta3, IL-6, 17beta-estradiol, or calcitonin) and was decreased by interferon-gamma, vitamin D3, and the antiinflammatory glucocorticoid dexamethasone. Changes in IL-8 secretion were paralleled by corresponding changes in IL-8 mRNA steady state levels. We conclude that hOC and hOCL synthesize and secrete high constitutive and inflammation-stimulated levels of the chemokine IL 8. Consequently, hOC-derived IL-8 could act as an important regulatory signal for bone, vascular, and immune cell recruitment and activation during normal and pathological bone remodeling. PMID- 9751521 TI - Dioxin perturbs, in a dose- and time-dependent fashion, steroid secretion, and induces apoptosis of human luteinized granulosa cells. AB - Dioxin (2,3,7,8-tetrachlorodibenzo-p-dioxin; TCDD) is the most toxic congener of a large class of environmental pollutants. Several studies have shown that TCDD exposure reduced fecundity and ovulatory rate in rats and increased the incidence of endometriosis in monkeys. Recent work suggests that TCDD's endocrine disrupting effects are, at least in part, caused by a direct action at the ovary. Although the factors involved in TCDD-induced toxicity are still under investigation, several studies have shown that TCDD induces programmed cell death, or apoptosis, in various tissues and may act in a similar fashion in the ovary. In the present study, we set out to evaluate the in vitro effects of TCDD on steroid secretion, specifically estradiol-17beta (E2) and progesterone, by human luteinized granulosa cells (LGC), and to further determine whether TCDD is capable of inducing apoptosis in this cell type. Human LGC were obtained from women participating in an in vitro fertilization program. Medium, with or without three different concentrations of TCDD and substrates [androstenedione (A4) or pregnenolone], was added to each culture. The media were collected at 4, 8, 12, 24, 36, and 48 h and were assayed by RIA. At 24 and 48 h, the LGC were fixed for assessment of DNA fragmentation via an in situ immunofluorescence technique. Transmission electron microscopy was also performed on LGC after 24 and 48 h with TCDD. TCDD, at all concentrations tested (3.1 pM, 3.1 nM, and 3.1 microM), significantly reduced E2 accumulation in the media at 8, 12, and 24 h, compared with controls. At 36 and 48 h, TCDD treatment (at 3.1 microM) caused a significant increase in E2, compared with controls. The effect of TCDD on E2 was abolished with the addition of A4. TCDD treatment did not alter progesterone accumulation. Apoptosis increased at 24 h with 3.1 microM TCDD, with no apparent effect at 3.1 nM. By 48 h, however, TCDD increased apoptosis in a dose-dependent manner. Transmission electron microscopy showed ultrastructural differences in LGC with 3.1 microM TCDD at 24 and 48 h. Collectively, the results of the present study suggest that TCDD perturbs E2 secretion by depletion of A4 precursor and increases apoptotic cell death of human LGC in a dose- and time-dependent fashion. PMID- 9751520 TI - Growth hormone promotion of tubulin polymerization stabilizes the microtubule network and protects against colchicine-induced apoptosis. AB - We have investigated the effect of GH on microtubular physiology in Chinese hamster ovary (CHO) cells stably transfected with the complementary DNA for the rat GH receptor (CHO-GHR(1-638)). We show here that after 30 min of human GH (hGH) treatment of CHO-GHR(1-638) cells, there was a significant increase in the level of polymerization of all four tubulin isoforms (alpha-, beta-, gamma-, and tyrosinated alpha-tubulin) compared with the serum-deprived state. However, this transient increase in the levels of polymerized tubulin after hGH treatment was particularly pronounced for beta- and tyr alpha-tubulin. For alpha- and gamma tubulin, the hGH-induced increase in polymerization state lasted to approximately 3 h and then declined by 7 h, whereas for beta- and tyr alpha-tubulin there was a decrease in the polymerization state at 1-2 h after hGH treatment compared with the level at 30 min (but still greater than the serum-deprived state) followed by a second but lesser wave of increased polymerization lasting to 7 h. The changes in the polymerization state of the tubulins were not accompanied by comparative changes in the level of total cellular tubulin. The proline rich box 1 region of the GH receptor was required for hGH to stimulate tubulin polymerization indicative that this event is JAK dependent. Increased tubulin polymerization still occurred in response to hGH in a receptor truncation lacking the carboxyl terminal half of the intracellular domain of the GH receptor indicative that hGH induced changes in intracellular calcium concentration is not required for tubulin polymerization. Prior treatment of CHO-GHR(1-638) cells with hGH retarded colchicine induced microtubule depolymerization and also prevented colchicine induced apoptotic cell death. The integrity of the microtubule network was not required for GH-induced STAT5 mediated transcription as treatment of cells with colchicine, vincristine, or vinblastine did not alter the fold stimulation of the STAT5 mediated transcriptional response to GH. Thus one consequence of cellular treatment with GH is alteration in microtubule physiology. PMID- 9751522 TI - Growth factor effects on apoptosis of rat gastric enterochromaffin-like cells. AB - Enterochromaffin-like (ECL) cells are histamine-containing endocrine cells in the gastric epithelium that show increased density during chronic atrophic gastritis. The current study determined cell number and apoptosis of isolated rat ECL cells in response to several growth factors. Isolated ECL cells from fundic mucosa (enrichment >90%) were grown in serum-free medium over 2-5 days. Cell number was determined by mitochondrial formazan production; apoptosis was measured by Tdt mediated dUTP nick end labeling reaction and DNA fragmentation-based enzyme linked immunosorbent assay. Immunocytochemistry and RT-PCR demonstrated the presence of epidermal growth factor receptor, neuronal growth factor receptor (type 1), and fibroblast growth factor (FGF) receptor (type 1). Gastrin (EC50, approximately 2 pM), transforming growth factor-alpha (TGF alpha; 10-30 ng/ml), and basic FGF (bFGF; 1-10 ng/ml) increased the total number of cultured ECL cells. bFGF augmented the gastrin (1 pM)-induced response. Beta-neuronal growth factor (10 ng/ml) and bFGF (2 ng/ml) decreased the programed death of ECL cells. Interleukin-1beta (100 pg/ml, 24 h) stimulated apoptosis 2- to 3-fold in ECL cells, and simultaneous incubation with TGF alpha (20 ng/ml) or bFGF (2 ng/ml) significantly inhibited this effect. ECL cells express specific receptors for gastrin, epidermal growth factor, neuronal growth factor, and FGF. bFGF prolonged ECL cell survival by inhibiting spontaneous apoptosis. Our data further indicate that TGF alpha and bFGF increase ECL cell number by inhibiting cytokine-induced programed cell death. PMID- 9751523 TI - Normalization of mineral ion homeostasis by dietary means prevents hyperparathyroidism, rickets, and osteomalacia, but not alopecia in vitamin D receptor-ablated mice. AB - 1,25-Dihydroxyvitamin D3 plays a major role in intestinal calcium transport. To determine what phenotypic abnormalities observed in vitamin D receptor (VDR) ablated mice are secondary to impaired intestinal calcium absorption rather than receptor deficiency, mineral ion levels were normalized by dietary means. VDR ablated mice and control littermates were fed a diet that has been shown to prevent secondary hyperparathyroidism in vitamin D-deficient rats. This diet normalized growth and random serum ionized calcium levels in the VDR-ablated mice. The correction of ionized calcium levels prevented the development of parathyroid hyperplasia and the increases in PTH messenger RNA synthesis and in serum PTH levels. VDR-ablated animals fed this diet did not develop rickets or osteomalacia. However, alopecia was still observed in the VDR-ablated mice with normal mineral ions, suggesting that the VDR is required for normal hair growth. This study demonstrates that normalization of mineral ion homeostasis can prevent the development of hyperparathyroidism, osteomalacia, and rickets in the absence of the genomic actions of 1,25-dihydroxyvitamin D3. PMID- 9751524 TI - Development of adrenal zonation in fetal rats defined by expression of aldosterone synthase and 11beta-hydroxylase. AB - The adult rat adrenal cortex is comprised of three concentric steroidogenic zones that are morphologically and functionally distinguishable: the zona glomerulosa, zona intermedia, and the zona fasciculata/reticularis. Expression of the zone specific steroidogenic enzymes, cytochrome P450 aldosterone synthase (P450aldo), and P450 11beta hydroxylase (P45011beta), produced by the zona glomerulosa and zona fasciculata/reticularis, respectively, can be used to define the adrenal cortical cell phenotype of these two zones. In this study, immunohistochemistry and in situ hybridization were used to determine the ontogeny of expression of P450aldo and P45011beta to monitor the pattern of development of the rat adrenal cortex. RIA was used to measure adrenal content of aldosterone and corticosterone, the resulting products of the two enzymatic pathways. Double immunofluorescent staining for both enzymes at gestational day 16 (E16) showed P45011beta protein expressed in cells distributed throughout most of the adrenal intermixed with a separate, but smaller, population of cells expressing P450aldo protein. Whereas expression of P45011beta protein retained a similar pattern of distribution from E16 to adulthood (ignoring distribution of SA-1 positive, presumptive medullary cells), P450aldo protein changed its pattern of distribution by E19, becoming localized in a discontinuous ring of cells adjacent to the capsule. By postnatal day 1, P450aldo protein distribution was similar to that observed in adult glands; P450aldo-positive cells formed a continuous zone underlying the capsule. In situ hybridization showed that the pattern of P45011beta messenger RNA expression paralleled protein expression at all times, whereas P450aldo messenger RNA paralleled protein at E19 and after, but was undetectable before E19. However, adrenal aldosterone and corticosterone, as measured by RIA, were detected by E16, supporting the functional capacity of both phenotypes for all ages studied. These data suggest that the development of the adrenal zona glomerulosa occurs in two distinct phases; initial expression of the glomerulosa phenotype in scattered cells of the inner cortex before E17, followed by a change in distribution to the outer cortex between E17 and E19. It is hypothesized that this change in distribution occurs via cell differentiation, rather than cell migration, and that a possible regulator of these events is the fetal renin-angiotensin system. PMID- 9751525 TI - Gastrin-producing endocrine cells: a novel source of histamine in the rat stomach. AB - Gastrin and histamine both potently stimulate secretion of acid into the gastric lumen. How these agents interact and how their release is controlled is poorly understood. Therefore, we decided to look for histamine in the antral portion of the rat stomach where the gastrin-producing G cells are located. We used immunocytochemical methods to visualize histamine, histidine decarboxylase (HDC, the enzyme that converts histidine to histamine), and the type 1 vesicular monoamine transporter (VMAT1, the protein responsible for moving histamine into vesicles for storage and release). We were surprised to find that histamine, HDC, and VMAT1 were all present in G cells. Our results suggest that G cells synthesize and secrete gastrin and histamine. Whether histamine acts in concert with gastrin to stimulate acid secretion, or functions as an autocrine inhibitor of gastrin release remains to be seen. PMID- 9751526 TI - An immunohistochemical study of Na+/I- symporter in human thyroid tissues and salivary gland tissues. AB - The human Na+/I- symporter (hNIS) is the plasma membrane protein that mediates active iodide uptake into several tissues, such as the thyroid and salivary glands. To study the distribution and cellular localization of the hNIS protein, we have generated a polyclonal antibody that could detect the hNIS protein by immunohistochemical staining on tissue sections. In normal thyroids, hNIS expression is heterogeneous, and it is only detected in sporadic thyrocytes of a given follicle. The hNIS protein was not detected in thyroid carcinomas, yet it was detected in the majority of thyrocytes in Graves' thyroids. In salivary glands, hNIS protein was not detected in acinar cells, but it was detected in ductal cells. The hNIS proteins are clustered in the basal and lateral membranes in cells stained positive for hNIS. PMID- 9751527 TI - Expression of growth hormone secretagogue-receptors by growth hormone-releasing hormone neurons in the mediobasal hypothalamus. AB - A novel class of synthetic compounds, termed GH-secretagogues (GHSs), have been shown to be potent stimulators of GH release, although their mechanism of action and functional significance remains obscure. The recent cloning of the rat GHS receptor (GHS-R) permitted the identification of numerous sites of expression of GHS-R in brain, but nothing is yet known about the cell types that express this receptor. We performed dual chromogenic and autoradiographic in situ hybridization to test the hypothesis that GHRH neurons in the hypothalamus coexpress GHS-R mRNA. GHS-R-hybridizing cells showed extensive overlap with GHRH expressing neurons in both the arcuate (Arc) and ventromedial (VMN) hypothalamic nuclei. Quantification of the double-labeled cells revealed that approximately 27% of GHRH-hybridizing neurons in the Arc, and 22% of those in the VMN, expressed the GHS-R gene. These studies are the first to colocalize the GHS-R to any neurochemical cell type in rat brain. The results provide evidence that the GHSs may directly modulate GHRH release, and thereby stimulate GH secretion, through interaction with the GHS-R on hypothalamic GHRH mRNA-containing neurons. PMID- 9751528 TI - A combination of osteoclast differentiation factor and macrophage-colony stimulating factor is sufficient for both human and mouse osteoclast formation in vitro. AB - Both human and murine osteoclasts can be derived in vitro from hematopoietic cells or monocytes that are co-cultured with osteoblasts or marrow-derived stromal cells. The osteoclastogenic stimulus provided by murine osteoblasts and marrow-derived stromal cells is now known to be mediated by osteoclast differentiation factor (ODF), a membrane-bound tumor necrosis factor-related ligand. This study demonstrates that mouse spleen cells and monocytes form osteoclasts when cultured in the presence of macrophage-colony stimulating factor (M-CSF) and a soluble form of murine ODF (sODF). Numerous multinucleated osteoclasts expressing tartrate resistant acid phosphatase (TRAP) and calcitonin receptor (CTR) formed within 7 days of culture and engaged in extensive lacunar bone resorption. Osteoclast number and bone resorption area was dependent on sODF concentration. Long-term cultured human monocytes also formed bone resorbing osteoclasts in response to co-stimulation by sODF and M-CSF, although this required more than 11 days in culture. This human osteoclast differentiation was strongly inhibited by granulocyte-macrophage colony stimulating factor. This study further characterises murine osteoclast differentiation caused by sODF and M-CSF co-stimulation in vitro, and shows that the same co-stimulation causes human osteoclast differentiation to occur. We propose that this methodology can be employed to investigate the direct effects of cytokines and other factors on human osteoclast differentiation. PMID- 9751529 TI - A C-terminal fragment of Agouti-related protein increases feeding and antagonizes the effect of alpha-melanocyte stimulating hormone in vivo. AB - Agouti-related protein (Agrp) is present in rat and human hypothalamus and is structurally related to agouti protein. Overexpression of either of these proteins results in obesity. However the effect of exogenous Agrp and its in vivo interaction with alpha-melanocyte stimulating hormone (alphaMSH), the likely endogenous melanocortin 3 and 4 receptor (MC3-R and MC4-R) agonist, have not been demonstrated. We report that 1 nmol of Agrp(83-132), a C-terminal fragment of Agrp, when administered intracerebroventricularly (ICV) into rats, increased food intake over a 24-h period (23.0+/-1.4 g saline vs 32.9+/-2.3 g Agrp, p<0.05). The hyperphagia was similar to that seen when 1 nmol of the synthetic MC3-R and MC4-R antagonist SHU9119 was given i.c.v. (19.6+/-1.8 g saline vs 32.5+/-1.7 g SHU9119, p<0.001). Both Agrp(83-132) and SHU9119 blocked the reduction in 1-h food intake of i.c.v. alphaMSH at the beginning of the dark phase. This effect occurred independently of whether the antagonists were administered simultaneously, or nine hours prior, to the alphaMSH. We have also shown Agrp(83-132) is an antagonist at the MC3-R and MC4-R, with similar inhibition of cAMP activation to that previously reported for the full length peptide. In conclusion, Agrp(83-132) administered i.c.v. increases feeding with long lasting effects and is able to inhibit the action of alphaMSH. This interaction may be mediated by the MC3-R and/or MC4-R. PMID- 9751530 TI - Variation among trauma centers' calculation of Glasgow Coma Scale score: results of a national survey. AB - BACKGROUND: Glasgow Coma Scale (GCS) scoring is enigmatic in intubated patients. To determine if there is consensus among Level I trauma centers, a national telephone survey was conducted. METHODS: Trauma registrars at state-verified or American College of Surgeons-verified Level I trauma centers were questioned about GCS scoring, recording, and reporting in patients who are intubated or intubated and pharmacologically paralyzed. RESULTS: Seventy-three centers were contacted. Seventy-one use initial GCS scores for registry recording. Intubated patients are given 1 point for verbal component plus eye and motor scores at 26% of centers and a total GCS score of 3 at 23%; GCS score is estimated with "T" given for verbal component at 16%, scored as unknown at 10%, always scored as 15 at 10%, and the method of scoring is unknown at 15%. Pharmacologically paralyzed intubated patients are given a total GCS score of 3 at 34%, GCS score is estimated with "T" given for verbal component at 18%, patients are given 1 point for verbal component plus eye and motor scores at 12%, scored as unknown at 11%, always scored as 15 at 8%, and the method of scoring is unknown at 16%. CONCLUSION: Wide variation in GCS scoring among Level I trauma centers was identified. Because GCS scores are used in treatment algorithms, trauma scoring, and outcome prediction (Trauma and Injury Severity Score), uniform scoring is essential and should be pursued. Use of state and national databases and outcome research may be adversely affected by the lack of consistent GCS scoring. PMID- 9751531 TI - Effect of hemorrhage on superior mesenteric artery flow during increased intra abdominal pressures. AB - BACKGROUND: Elevations in intra-abdominal pressure (IAP) adversely affect organ function. Prior hemorrhage and resuscitation exacerbates the cardiac and pulmonary effects of IAP. We have recently shown that superior mesenteric artery flow (SMAF) is reduced with increasing IAP. This study was designed to determine whether and how hemorrhage and resuscitation affects SMAF with increasing IAP. METHODS: Ten pigs were divided into two groups after placement of a Doppler flow probe around the proximal SMA and insertion of a pulmonary artery (PA) catheter. Group 1 underwent intraperitoneal infusion of fluid to increase IAP to 10, 20, 30, and 40 mm Hg followed by a 20-minute equilibration period at each IAP. Group 2 was hemorrhaged 20% of circulating volume followed by standard resuscitation. After equilibration, this group had IAP increased in the same manner as group 1. Cardiac output (CO), PA pressures, and SMAF were recorded 1 hour after laparotomy and after equilibration at each IAP. Comparisons were made using repeated measures of analysis of variance, Student's t test, and linear regression analysis. RESULTS: In group 2, a reduction in SMAF was noted at 30 and 40 mm Hg of IAP when compared with baseline (p = 0.009). This reduction was not seen in group 1. There was also a significant (p = 0.001) reduction in CO between baseline and all levels of increased IAP in group 2. This decrease was again not seen in group 1. The correlation between CO and SMAF in group 2 was r = 0.74, r2 = 0.55, p = 0.0001. There was no significant correlation between CO and SMAF in group 1. CONCLUSION: SMAF and CO are reduced with increasing IAP to a greater degree when preceded by hemorrhage and resuscitation. Although there is a strong correlation between the reductions in CO and SMAF, the reduction in SMAF is greater than the reduction in CO. This finding suggests that optimizing cardiac function alone during periods of even moderate levels of increased IAP may be inadequate to normalize SMAF. This lends further support for early abdominal decompression in the treatment of trauma patients with increased IAP. PMID- 9751532 TI - P-selectin blockade is beneficial after uncontrolled hemorrhagic shock. AB - OBJECTIVES: The selectins play an important role in the neutrophil-mediated injury after hemorrhagic shock and resuscitation. The aim of this study was to investigate the effect of P-selectin blockade after HS and resuscitation. METHODS: Forty-eight Sprague-Dawley rats were subjected to controlled combined with uncontrolled HS and resuscitation. Rats were divided into three groups (n = 16/group): (1) sham, no HS; (2) control, HS + resuscitation + drug vehicle; (3) treated, HS + anti-P-selectin monoclonal antibody. Transaminase levels to measure hepatocellular injury, liver myeloperoxidase to assess neutrophil infiltration, and histology were analyzed and compared between groups. Survival was followed for 3 days and was compared statistically. RESULTS: Survival significantly increased from 30% in the control group to 70% in the treated group. Hepatocellular and structural injury as well as neutrophil infiltration were significantly decreased in treated animals. CONCLUSION: Blockade of P-selectin resulted in decreased hepatocellular injury and increased survival in our model of uncontrolled HS. Selectins may be important therapeutic targets for blockade in the treatment of HS. PMID- 9751533 TI - Determining anatomic injury with computed tomography in selected torso gunshot wounds. AB - BACKGROUND: Changes in the management of torso gunshot wounds (TGSWs) have evolved in recent years as a result of differences between military and civilian injuries and increasing interest in avoiding nontherapeutic invasive procedures. The objective of this study was to establish the utility and accuracy of computed tomography (CT) in the evaluation of selected patients with TGSWs. METHODS: Retrospective review for a 6-year period of patients who sustained TGSWs and underwent CT solely for the purpose of trajectory determination. Patients had complete physical examinations and plain radiographic evaluations by a dedicated group of in-house trauma surgeons. When trajectory was indeterminate after evaluation, CT was performed. In some cases, CT was used when trajectory was determined to be intracavitary but organ injury was believed to be unlikely or amenable to nonoperative management. RESULTS: Fifty TGSW patients underwent 52 computed tomographic scans. Abdominal/pelvic CT was performed in 37 patients, and thoracic CT was performed in 15 patients. All patients were stable and none sustained complications attributable to CT or delay in therapy. Twenty of 37 abdominal/pelvic computed tomographic scans excluded transabdominal or pelvic trajectory. Seventeen of 37 scans proved transabdominal or pelvic trajectory; nine laparotomies were performed, and eight patients were observed. Nine of 15 thoracic computed tomographic scans excluded transmediastinal trajectory. Six of 15 scans suggested vascular proximity and prompted further workup, which was positive in two cases. CONCLUSION: CT of selected TGSW patients is safe and may reduce the incidence of invasive diagnostic procedures. A prospective evaluation of CT for TGSW patients is warranted. PMID- 9751534 TI - Effect of alpha(alpha)-cross-linked hemoglobin and pyridoxalated hemoglobin polyoxyethylene conjugate solutions on gastrointestinal regional perfusion in hemorrhagic shock. AB - BACKGROUND: Hemoglobin-based blood substitutes may cause vasoconstriction, which could limit organ perfusion during trauma resuscitation. We investigated the effect of two hemoglobin solutions on regional blood flow and mucosal perfusion in the gastrointestinal tract in a hemorrhagic shock model. METHODS: Twenty-four swine were bled 30% of blood volume over 1 hour. Six additional animals were anesthetized and monitored but did not undergo hemorrhage. Bled animals were resuscitated with alpha(alpha)-hemoglobin (alpha(alpha)Hb), pyridoxalated hemoglobin polyoxyethylene conjugate (PHP), shed blood, or lactated Ringer's solution. Regional blood flow was measured by radiolabeled microspheres. Gastric mucosal perfusion was estimated by measuring intramucosal pH (pHi) by tonometry. RESULTS: PHP and shed blood restored small-bowel flows to sham values, whereas lactated Ringer's solution and alpha(alpha)Hb did not. Shed blood and PHP, but not alpha(alpha)Hb, restored cardiac index (CI) to baseline (p < 0.05). Mean pulmonary artery pressure was elevated over baseline with alpha(alpha)Hb and PHP and remained elevated with alpha(alpha)Hb (p < 0.05). pHi was significantly lower after resuscitation with PHP than with other fluids. CONCLUSION: PHP was efficacious in restoring CI and small-bowel flow, but the pHi remained low, indicating possible continued mucosal ischemia. Alpha(alpha)Hb led to limited recovery of CI and small-bowel blood flow but restored pHi close to baseline. Shed blood was efficacious in restoration of pHi, gastrointestinal blood flows, and systemic hemodynamics. PMID- 9751535 TI - Redefining cardiovascular performance during resuscitation: ventricular stroke work, power, and the pressure-volume diagram. AB - OBJECTIVES: (1) To compare left ventricular stroke work index (SW) and left ventricular power output (LVP), hemodynamic variables that encompass blood pressure as well as blood flow, with the purely flow-derived hemodynamic and oxygen transport variables as markers of perfusion and outcome in critically injured patients during resuscitation. (2) To use the ventricular pressure-volume diagram to define characteristic hemodynamic patterns in the determinants of SW and LVP that are associated with survival. METHODS: This was a cohort study at a university Level I trauma center during the course of 1 year. A consecutive series of patients was monitored with a volumetric pulmonary artery catheter during the initial 48 hours of resuscitation. Heart rate, SW, LVP, cardiac index, and oxygen delivery and consumption during resuscitation were compared using multivariate logistic regression analysis with regard to the ability to clear lactate in less than 24 hours and survival. Receiver operating characteristic curves were constructed to determine threshold values for SW and LVP. Ventricular pressure-volume diagrams were used to describe characteristic patterns in the determinants of SW and LVP in survivors and nonsurvivors. Preload was expressed as left ventricular end-diastolic volume index, afterload as aortic input impedance (Ea), and contractility as ventricular end-systolic elastance (Ees). The ratio of Ea/Ees (RATIO) was used as a measure of ventricular-arterial coupling, which describes the efficacy of energy transfer from the heart to the vascular system. RESULTS: One hundred eleven patients (87 survivors, 24 nonsurvivors) met study criteria. Survivors had a significantly higher SW (4,510 +/- 1,070 vs. 3,440 +/- 980 mm Hg x mL x m(-2); p < 0.0001) and LVP (370 +/- 94 vs. 270 +/- 81 mm Hg x L x min(-2) x m(-2); p < 0.0001) than nonsurvivors. Heart rate, SW, and LVP were the only studied variables that were significantly related to lactate clearance and survival by logistic regression. Threshold values determined by the receiver operating characteristic curves were 4,000 mm Hg x mL x m(-2) for SW and 320 mm Hg x L x min(-1) x m(-2) for LVP. Survivors had better ventricular-arterial coupling than nonsurvivors, indicated by a lower RATIO (0.32 +/- 0.22 vs. 0.54 +/- 0.38; p = 0.003). This lower RATIO was attributable to lower levels of Ea (2.7 +/- 0.7 vs. 3.4 +/- 0.8 mm Hg x mL(-1) x m(-2); p = 0.0003) and a trend toward higher levels of Ees (13 +/- 11 vs. 9.9 +/- 7.3 mm Hg x mL(-1) x m(-2); p = 0.12). CONCLUSION: Thermodynamic perfusion variables that encompass both pressure and flow, such as SW and LVP, are more closely related to perfusion and outcome than the purely flow-derived variables. The higher SW and LVP in survivors is related to better ventricular-arterial coupling, and therefore more efficient cardiac function. Cutoff values for LVP of 320 mm Hg x L x min(-1) x m(-2) and for SW of 4,000 mm Hg x mL x m(-2) may be useful thresholds for evaluating hemodynamic performance during resuscitation. PMID- 9751536 TI - Effects of short heat exposure on human red and white blood cells. AB - BACKGROUND: The infusion of warm intravenous fluid (IVF) is a simple and effective method used to maintain or restore core body temperature. At present, 40 degrees C is believed to be the highest temperature that can be safely administered. There is concern that temperatures greater than 40 degrees C may harm blood cells. The mixing time of IVF infused into a high-flow vein such as the superior vena cava is very short, however, approximately 300 milliseconds. We will determine the maximum temperature and exposure time tolerated by human red and white blood cells without producing injury. METHODS: Whole blood and isolated neutrophils were exposed to temperatures (40-80 degrees C) for short time intervals (150-1,200 milliseconds). Lethal injury to red and white blood cells was measured by the plasma free hemoglobin and percent viability, respectively. Neutrophil viability was measured by trypan blue staining. Sublethal injury to red and white cells was measured by osmotic fragility and oxidative burst, respectively. Neutrophil oxidative burst was measured by chemiluminescence. Control values were compared with postexposure values using analysis of variance with p < 0.05 indicating significance. RESULTS: Lethal injury to red blood cells did not occur until exposure at 70 degrees C for 300 milliseconds (plasma free hemoglobin, 116.3 +/- 34.7 mg%; p < 0.05). Lethal injury to neutrophils did not occur, even at exposure at 80 degrees C for 1,200 milliseconds. Sublethal injury to red blood cells did not occur until exposure at 60 degrees C for 1,200 milliseconds. Sublethal injury to neutrophils did not occur until exposure at 60 degrees C for 600 milliseconds (percent change in oxidative burst = 28.9 +/- 0.96%; p < 0.05). CONCLUSIONS: The exposure of human red blood cells and neutrophils to temperatures up to 60 degrees C for up to 600 milliseconds does not cause lethal or sublethal injury. These findings contribute to the body of evidence supporting the use of centrally infused IVF at temperatures greater than 40 degrees C for active core rewarming. PMID- 9751537 TI - Lower limb trauma caused by power-driven cultivators: report of 23 cases. AB - OBJECTIVE: To determine the mechanism and the severity of injuries caused by power-driven cultivators. METHODS: This retrospective study analyzed the clinical records of 20 patients treated from 1984 to 1996 for a total of 23 lower limb injuries caused by power-driven cultivators (three patients had bilateral injuries) in the Nice University Hospital. RESULTS: A total of 90% of the accidents occurred when the machine was put into reverse and the limb was caught by the rotary blades; the cause of the remaining accidents was unknown. Of the 23 patients, 10 patients (43.5%) suffered posterior dislocation of the knee due to forced hyperextension. Injuries were classed in two groups as a function of their prognosis: group I consisted of osteomuscular lesions without vascular or nerve involvement (11 lower limbs, 11 patients). The mortality rate in this group was 9%, the rate of major amputation was 18%, and the prognosis was favorable in 82% of the cases. Group II corresponded to lower limb injuries with neurovascular involvement (12 lower limb injuries in 10 patients: one patient belonged to both group I and group II). Acute lower limb ischemia was constant in group II; the mortality rate was 20% (two of 10 patients), and the rate of major amputation was 41.6% (five of 12 patients; three emergency amputations and two secondary amputations). CONCLUSION: These agricultural machines can cause severe trauma, and the resulting wounds are contaminated by telluric germs in rural areas. Paradoxically, power-driven cultivators are not legally classified as "dangerous machines." Modification of existing legislation in this field would seem advisable. PMID- 9751538 TI - Trauma patients with multiple extremity injuries: resource utilization and long term outcome in relation to injury severity scores. AB - BACKGROUND: Trauma patients with multiple extremity injuries (MEI) make heavy demands on hospital resources and face long-term difficulties in rehabilitation, yet the literature contains little about their treatment as a distinct group. METHODS: In this study, a cohort of 54 patients with MEI, all treated at a Level I trauma center, was compared with a trauma control (TC) group that had major injuries not focused at the extremities (but excluding patients with neurologic sequelae of head or spinal cord injuries). Demographic features, primary measures reflecting utilization of hospital resources, return-to-employment and productivity data, and health-related quality of life scores (Medical Outcomes Study 36-Item Short Form Health Survey [SF-36]) were compared. RESULTS: Although mean Injury Severity Scores (ISS) for the MEI and TC groups were almost identical (16.2 and 17.4, respectively), the patients with MEI had a mean hospital stay almost twice as long (25 vs. 13 days) and had double the resource intensity weight compared with the TC group. After discharge, the trend of the MEI group was to greater long-term disability, based on SF-36 scores, and lower "return to productivity" figures. The ISS did not predict the greater demands on resources made by the MEI group relative to our TC group. Main injury severity scores for the extremities were more predictive than the ISS for length of hospital stay and the SF-36 concepts at the 2-year follow-up evaluation. CONCLUSION: The study emphasizes the need for injury scoring systems that better predict the needs of patients with MEI and that will serve as a basis for equitable funding of trauma centers. PMID- 9751539 TI - Experimental arrow wounds: ballistics and traumatology. AB - OBJECTIVE: To provide information on the ballistics and the wounding potential of different arrows or bolts fired from different weapons and to investigate the suitability of simulant media for experimental arrow wounds. METHODS: A longbow, a compound bow, and a crossbow were used to fire a variety of modern and ancient arrows. Fresh corpses of four adult pigs (47 shots) and blocks of gelatin and soap (48 shots) were used as target media, and the resulting wound tracts were examined. The range of fire was 8 m and the velocity was recorded at a distance of 3 m (and 16 m in additional shots) by light screen devices. RESULTS: The mean velocities recorded ranged from 45 m/s (longbow) to 67 m/s (compound bow). The excellent exterior ballistics of arrows results in only a small initial decrease in velocity of O.10 to 0.18 m x s(-1) x m(-1). The penetration depths were reproducible for the same arrowhead fired into the same simulant medium but differed considerably when compared with those in soft tissue. In nonbone tissue, the penetration depth was substantial (17-60 cm) and depended on velocity and especially on the type of arrowhead. All arrows penetrated deeply into the large body cavities and injured organs as long as no thick bone had to be perforated. Flat bones such as ribs were always perforated. Extraction of arrowheads from thick bone proved to be difficult in some cases. The wounding mechanism was a combination of incision and puncture, which facilitated deep penetration of tissue and produced clean-cut wounds. CONCLUSION: Gelatin and soap are not suitable for experimental arrow wounds. Every arrow wound carries a lethal potential. The severity of the wound depends primarily on the target area and the type of arrowhead. Extraction of arrowheads from thick bone has to be performed carefully. PMID- 9751540 TI - Spinal cord injury incidence in Mississippi: a capture-recapture approach. AB - BACKGROUND: Many studies have investigated spinal cord injury incidence rates. Few, however, have adjusted for the underascertainment. The current study used the capture-recapture method to estimate the ascertainment-corrected spinal cord injury incidence rate in Mississippi. METHODS: Two sources were used for case ascertainment: Mississippi's spinal cord injury registry and hospital reports. The two-sample capture-recapture method was used to adjust for undercount. RESULTS: Two hundred one spinal cord injuries were reported to or found by the Mississippi State Department of Health in 1993, with a crude incidence rate of 7.8 per 100,000 population per year among hospital admissions and prehospital fatalities. Using the two-sample capture-recapture method, it is estimated that the incidence rate would be 9.3 per 100,000 population per year. CONCLUSION: Capture-recapture estimates suggest that Mississippi's spinal cord injury incidence rate is more than twice the national average. PMID- 9751541 TI - Percutaneous computed tomographic-controlled ventriculostomy in severe traumatic brain injury. AB - OBJECTIVE: Percutaneous computed tomographic (CT)-controlled ventriculostomy (PCV) was introduced for the monitoring of intracranial pressure in patients with severe traumatic brain injury who did not require simultaneous decompressive trepanation. METHODS: PCV (n = 14) was compared with conventional burr hole ventriculostomy (n = 13) based on prospectively collected data. RESULTS: PCV proved to be a successful technique in all cases and also when a burr hole ventriculostomy was impossible previously. There were no complications. In burr hole ventriculostomy, there were one unsuccessful insertion and one catheter contamination. The main advantage of PCV over burr hole ventriculostomy was a significant (p < 0.05) reduction in the time required to perform the procedure. In ventriculostomy directly after the initial evaluation in the emergency department, the operation time was reduced from 45 +/- 11 to 22 +/- 14 minutes. The interval between cranial computed tomography and start of operation was reduced from 78 +/- 38 to 33 +/- 12 minutes, and between initial cranial computed tomography and intensive care unit admittance, from 138 +/- 37 to 73 +/- 28 minutes. For patients requiring ventriculostomy while being treated in the intensive care unit, the duration of the procedure (i.e., absence from the intensive care unit) was able to be reduced from 111 +/- 24 to 81 +/- 21 minutes. CONCLUSION: Distinct time savings are the major advantages of PCV, allowing exact catheter positioning even with very narrow ventricles. PMID- 9751542 TI - Treatment of clavicular aseptic nonunion: comparison of plating and intramedullary nailing techniques. AB - OBJECTIVE: The aim of this retrospective study was to investigate and compare the effects of plating and intramedullary nailing in the treatment of clavicular aseptic nonunion. METHODS: Thirty-three consecutive patients with middle-third clavicular aseptic nonunions with previous nonoperative treatment were treated by plating (13 patients) and intramedullary nailing (20 patients) with supplementary cancellous bone grafting. The indications for such treatment were middle-third aseptic nonunions without previous operative treatment and with local pain or tenderness, deformity, or neurologic complaint. The choice of plating or intramedullary nailing was according to the surgeon's individual preference. RESULTS: Twenty-nine patients were followed for at least 1 year (range, 1-7 years; median, 3 years; plating, 11; intramedullary nailing, 18). The union rate was 81.8% (9 of 11) for plating and 88.9% (16 of 18) for intramedullary nailing (p = 0.35, Fisher's exact test). The union period was 4.0 +/- 1.3 months for plating and 4.1 +/- 1.1 months for intramedullary nailing (p = 0.80, unpaired Student's t test). The complication rate was 27.3% (3 of 11) for plating and 11.1% (2 of 18) for intramedullary nailing (p = 0.21, Fisher's exact test). There were no significant differences in other parameters. CONCLUSION: Intramedullary nailing may have a higher union rate with a lower complication rate than plating (p > 0.05). At least in common situations, it is not inferior to plating. Whenever possible, therefore, intramedullary nailing should be used preferentially to treat clavicular aseptic nonunion without previous operative treatment. Nevertheless, both techniques have relatively higher nonunion rates in the treatment of clavicular nonunion than in the treatment of other long-bone nonunions. Gentle handling of surrounding soft tissues to reduce bony fragments should be strictly executed. PMID- 9751543 TI - Improved results in treatment of femoral shaft fractures with the unreamed femoral nail? A multicenter experience. AB - OBJECTIVES: The first studies of intramedullary nailing with the Arbeidsgemeinschaft fur Osteosynthesefragen (AO) unreamed femoral nail in selected clinics showed favorable results. Daily practice, however, is that femoral fractures are treated in a variety of clinics by a mixture of surgeons. To evaluate whether similar results could be obtained in general practice, a prospective multicenter trial was undertaken, involving a variety of university and general hospitals in one country. METHODS: Between August of 1994 and June of 1996, 122 patients with 129 traumatic femoral shaft fractures treated with the unreamed femoral nail in eight hospitals were included in this study. Patients who had a reoperation with an unreamed femoral nail or patients with a pathologic fracture of the femur were excluded from this part of the study. Of these patients, 58 patients had multiple injuries, and 33 of the fractures had open soft-tissue injury. RESULTS: Postoperative infection occurred in four patients; the nail broke in one patient. In total, nine patients (6.6 %) sustained general complications, five of which developed adult respiratory distress syndrome (3.6%). Non-union occurred in seven patients (5.1%) and delayed union occurred in four cases (2.9%) with a reintervention rate of 6.6%. CONCLUSION: In this study, a decrease in the number of patients who develop adult respiratory distress syndrome through the use of a thin unreamed nail could not be demonstrated. The promising early callus formation and good consolidation mentioned in previous studies could not be confirmed. We find that the technical and clinical results in this study of unreamed femoral nailing in a mixture of clinics and by a variety of surgeons are comparable to the results of reamed nailing in the literature and are not as favorable as in the previous reports. PMID- 9751544 TI - Factors contributing to delayed extremity amputation in burn patients. AB - BACKGROUND: Previous series of traumatic amputations have noted that delay in amputation results in prolonged hospital stay and delayed rehabilitation. A series of major extremity amputations after burn injury was analyzed to identify the frequency of delayed amputation and to identify factors resulting in the delay. METHODS: Chart review of burn admissions between January of 1991 and December of 1995. RESULTS: Twenty-eight patients underwent a total of 44 major extremity amputations. Thirty-five amputations in 22 patients were performed by postburn day 16 (mean 4.3). Nine amputations in six patients were delayed beyond postburn day 26 (mean, 48.3). Delayed amputations occurred in the subgroups of deep thermal burns with extensive necrosis and thermal burns complicated by infections. Early amputation was associated with a 13.6% mortality rate, delayed amputation with a 50% mortality rate. CONCLUSION: There is a bimodal distribution of time to amputation determined by mechanism of injury, severity of burn, and infectious complications. Earlier identification of nonsalvageable limbs may decrease infectious complications and improve the chances of patient survival. PMID- 9751545 TI - Acute metabolic and endocrine effects of induced hypothermia in hemorrhagic shock: an experimental study in the pig. AB - BACKGROUND: Hypothermia is considered harmful in trauma patients. In surgery, hypothermia is occasionally used to reduce metabolism and protect the brain. Recent studies in animals have also shown protective effects of hypothermia in hemorrhagic shock. The aim of this study was to evaluate the metabolic and endocrine effects of induced hypothermia in hemorrhagic shock. METHODS: Half of the individually calculated blood volume was removed from 17 anesthetized piglets. They were then randomized to normothermia or hypothermia and followed for 4 hours after hemorrhage. RESULTS: In the hypothermic pigs, arterial PO2 increased from 10.3 +/- 0.7 to 16.4 +/- 0.9 kPa, but it remained unchanged in the normothermic group. The serum levels of potassium increased from 3.9 +/- 0.2 to 5.0 +/- 0.2 mmol/L in the normothermic group. In the hypothermic pigs, the potassium levels temporarily decreased from 3.8 +/- 0.1 to 3.0 +/- 0.1 mmol/L but then returned to baseline levels. The levels of serum catecholamines surged in both groups during hemorrhage. They remained elevated in normothermic pigs but declined in the hypothermic group. CONCLUSION: In porcine hemorrhagic shock, induced hypothermia increases arterial oxygen tension and stabilizes serum levels of potassium and catecholamines. PMID- 9751546 TI - Relative bradycardia in patients with traumatic hypotension. AB - BACKGROUND: Tachycardia is considered a physiologic response to traumatic hypotension. The inability of the heart to respond to shock with tachycardia has been described as paradoxical bradycardia or relative bradycardia. The incidence and clinical significance of this condition in major trauma is not known. The objective of this study was to examine the incidence and prognostic significance of tachycardia and relative bradycardia in patients with traumatic hypotension. Relative bradycardia is defined as a systolic pressure < or = 90 mm Hg and a pulse rate < or = 90 beats per minute. METHODS: This is a retrospective study conducted at a large Level I academic trauma center during a 4-year period. Seventeen demographic and injury severity factors were analyzed for their possible role in tachycardic or bradycardic response in hypotensive patients. Incidence and mortality were derived for each subpopulation. Bivariate analysis of the association of incidence and mortality with each risk factor was performed. Factors with p values < 0.2 were included in stepwise logistic regression analyses that identified significant risk factors and derived adjusted relative mortality risks between tachycardic and bradycardic hypotensive patients. RESULTS: Excluding transfers and patients dead on arrival, 10,833 major trauma patients were seen during the study period. Seven hundred fifty patients (6.9%) had systolic blood pressure < or = 90 mm Hg; 533 patients had tachycardia (overall incidence of 4.9%, or 71.1% of hypotensive patients), and 217 patients had bradycardia (overall incidence of 2.0%, or 28.9% of hypotensive patients). The overall crude mortality was 29.2% among tachycardia patients and 21.7% among bradycardia patients (crude relative risk = 1.34; 95% confidence interval = 1.00 1.81; p = 0.047). The adjusted relative mortality risk between the two groups was 1.23 (95% confidence interval = 0.84-1.73; p = 0.284). Multivariate analysis showed that patients with relative bradycardia in the subgroups with Injury Severity Scores > or = 16, chest Abbreviated Injury Scale scores > or = 3, or abdominal Abbreviated Injury Scale scores > or = 3 had significantly better survival than patients with similar injuries presenting with tachycardia. CONCLUSION: Relative bradycardia in hypotensive trauma patients is a common hemodynamic finding. Mortality among tachycardic patients was more predictable than among bradycardic patients using commonly used demographic and injury indicators. The presence of relative bradycardia in some subgroups of patients with severe injuries seems to be associated with better prognosis than the presence of tachycardia. PMID- 9751547 TI - Cell count ratio: new criterion of diagnostic peritoneal lavage for detection of hollow organ perforation. AB - OBJECTIVES: Diagnostic peritoneal lavage (DPL) had been widely used in evaluating patients with suspected intraperitoneal injuries due to its high sensitivity. If the positive criteria are strictly followed, however, the incidence of nontherapeutic laparotomies will be unacceptably high. This realization has become more important recently with the popularization of nonoperative treatment for blunt solid organ injuries. For these patients, the early diagnosis of an associated hollow organ perforation is mandatory. METHODS: Three hundred and twenty patients undergoing DPL over an 18-month period were retrospectively reviewed to evaluate the usefulness of "cell count ratio" in diagnosing hollow organ perforation. The cell count ratio was defined as the ratio between white blood cell count and red blood cell count in the lavage fluid divided by the ratio of the same parameters in the peripheral blood. RESULTS: Two hundred twelve patients were diagnosed as having a positive DPL according to the classic criteria. Forty-four patients (21%) had a cell count ratio of greater than or equal to 1. The diagnosis at laparotomy was small bowel perforation in 31 patients, colon perforation in eight patients, diaphragmatic hernia in one patient, pancreatic transection in two patients, and liver laceration in two patients. None of the patients with a cell count ratio of less than I sustained hollow organ perforation. The average interval from injury to DPL was 5 hours, with the shortest being 1.5 hours. CONCLUSION: A cell count ratio of greater than or equal to 1 predicted hollow organ perforation with a specificity of 97% and a sensitivity of 100%. The selective use of the cell count ratio has improved the probability of early diagnosis of bowel perforation without increasing the cost of care. Nonoperative management can be applied more confidently to those patients sustaining a blunt solid viscus injury of the abdomen if the cell count ratio is low. We conclude that the cell count ratio of DPL effluent is a very sensitive and specific indicator of hollow organ perforation. In the treatment of blunt abdominal injuries, if the cell count ratio is positive, nonoperative treatment should be abandoned and a laparotomy undertaken. PMID- 9751548 TI - Outcome after hemorrhagic shock in trauma patients. AB - BACKGROUND: It is essential to identify patients at high risk of death and complications for future studies of interventions to decrease reperfusion injury. METHODS: We conducted an inception cohort study at a Level I trauma center to determine the rates and predictors of death, organ failure, and infection in trauma patients with systolic blood pressure < or = 90 mm Hg in the field or in the emergency department. RESULTS: Among the 208 patients with hemorrhagic shock (blood pressure < or = 90 mm Hg), 31% died within 2 hours of emergency department arrival, 12% died between 2 and 24 hours, 11% died after 24 hours, and 46% survived. Among those who survived > or = 24 hours, 39% developed infection and 24% developed organ failure. Increasing volume of crystalloid in the first 24 hours was strongly associated with increased mortality (p = 0.00001). CONCLUSION: Hemorrhage-induced hypotension in trauma patients is predictive of high mortality (54%) and morbidity. The requirement for large volumes of crystalloid was associated with increased mortality. PMID- 9751549 TI - Putting a lid on injury costs: the economic impact of the California motorcycle helmet law. AB - BACKGROUND: This study analyzed the effect of California's motorcycle helmet law on injury costs. METHODS: An economic evaluation was performed using state hospital discharge data, county-level cost data, and statewide crash reports to estimate the costs, charges, and lost productivity from motorcycle injuries. Total and per person costs and changes in these costs were estimated. RESULTS: Total medical care costs were $35 million less in 1993 than in 1991, a reduction of 35%. Costs decreased for all payer categories, and 73% of the reduced hospitalization costs were attributable to reduced costs for patients with head injuries. Initial hospital costs for patients with head injuries were $18,527 compared with $10,350 for patients without head injuries. CONCLUSION: During the first 2 years of implementation of California's helmet law, there were reduced costs for injuries and fatalities and large dollar savings to the state and other payers compared with the previous year. PMID- 9751550 TI - Traumatic renal artery occlusion: a 15-year review. AB - BACKGROUND: To better define what constitutes appropriate treatment for traumatic renal artery occlusion, we report our 15-year experience in managing this injury. METHODS: A retrospective chart review was performed to evaluate treatment outcomes and complications of 12 patients (13 injuries) who presented to our trauma centers with renal artery occlusion secondary to blunt injury. RESULTS: Five of 12 patients underwent attempted surgical revascularization with a median warm ischemia time of 5 hours (range, 4.5-36 hours). Of these five patients, one required nephrectomy for inability to establish arterial flow, three demonstrated no function, and one had return to 9% differential function on postoperative renal scan. Seven patients did not have attempted revascularization, and none of them experienced immediate complications. Hypertension developed in three patients (43%) who required nephrectomy to control blood pressure at a mean of 5 months after injury (range, 3-7 months). Four patients remained asymptomatic and normotensive at a mean follow-up of 11 months (range, 4 weeks to 2.6 years). CONCLUSION: Surgical revascularization for traumatic renal artery occlusion seldom results in a successful outcome. Patients who are observed must have close follow-up for hypertension. PMID- 9751551 TI - Stable pediatric blunt trauma patients: is trauma team activation always necessary? AB - BACKGROUND: An increasing number of studies on adult trauma patients have questioned the need for trauma team activation for stable patients dictated only by mechanism of injury. This triage approach seems to burden the limited resources of the trauma center and may prove to be cost-ineffective. The objective of our study was to determine the predictive value and the sensitivity and specificity of blunt injury mechanism for major trauma in stable pediatric trauma patients. METHODS: Patients 0 to 14 years old injured by injury mechanisms modified from the American College of Surgeons trauma triage criteria and presenting to our American College of Surgeons-verified regional pediatric trauma center from the field between July 1, 1993, and July 31, 1994, were included. Physiologically and anatomically stable patients were identified and subgroup analysis was performed to determine the negative and positive predictive value and sensitivity, and the specificity of blunt injury mechanisms for major trauma [Injury Severity Score > 15] in this group. RESULTS: One hundred ninety-four patients met the study criteria. One hundred forty-three patients (73.6%) had trauma team activation only for mechanism of injury. Of these patients, four patients had Injury Severity Score > 15. The positive and negative predictive values of injury mechanisms modified from the American College of Surgeons trauma triage criteria were 2.8% and 90.2%, respectively, for major trauma in stable pediatric blunt trauma patients. The sensitivity and specificity were 44.4% and 24.9%, respectively. CONCLUSION: Mechanisms of injury seem to have limited value as predictors of injury severity in stable pediatric blunt trauma patients. A modified response level for these patients may prove to be a safe and practical alternative to current practice. PMID- 9751552 TI - Creating injury episodes using medical claims data. AB - BACKGROUND: Health care episodes are traditionally created for a specific condition using defined relevant diagnosis and procedure codes and a start and end period. Our goal is to use 1987 to 1989 medical claims data to create distinct episodes of care as a result of injury. METHODS: Claims for 102,000 people younger than 65 years were obtained from Medstat Systems, Inc. Injury claims were identified by International Classification of Diseases, 9th Revision, Clinical Modification diagnosis codes and separated into 10 body regions. Using linked inpatient and outpatient claims data, we established clear zones--a maximum period for a return visit for medical treatment--for each of 10 body regions injured by hospitalization status. These clear zones were used to create episodes of injury. RESULTS: A total of 295,165 injury claims created 79,564 episodes of injury. Limb and trunk injuries typically have the most follow-ups in terms of number of claims and spacing between claims. Brain injuries, even for admitted patients, result in an average of fewer than two follow-up claims. On average, hospitalized patients require only one more follow-up than nonadmitted patients. CONCLUSIONS: This paper presents a method for identifying injury episodes using a medical claims database. The analysis suggests that follow-up to check for minor long-term sequelae of brain injury is rare. PMID- 9751553 TI - Posterior interosseous flap and its variations for coverage of hand wounds. AB - BACKGROUND: Conventional posterior interosseous flap has the disadvantage of partial or even complete necrosis of the flap when there is anatomical variation or contusion around its distal pedicle. To make it a more reliable flap, three types of auxiliary procedures were designed. METHODS: (1) When there is congestion after inset of the distally based flap, an additional venous anastomosis was carried out. (2) When there is anatomical variation so that a distally based flap could not be raised without compromising the nerve branches, or when contusion was found around the distal pedicle, the flap was changed into a free flap. This design is also indicated for coverage of the distal fingers. (3) When the patient is elderly with possible peripheral arterial disease, the flap was raised with a wide base, incorporating the branches of both the anterior and posterior interosseous arteries. There were eight, 36, and five patients in each group, respectively. RESULTS: There was only one failure in the free flap group. No partial necrosis of the flap was found. Other complications were analyzed. CONCLUSION: With these backup procedures, the posterior interosseous flap can be more widely used with safety. By combining various reconstructive armaments, the result of a conventional procedure can be improved. PMID- 9751554 TI - Braced for impact: reducing military paratroopers' ankle sprains using outside the-boot braces. AB - BACKGROUND: Ankle injuries account for 30 to 60% of all parachuting injuries. This study was designed to determine if outside-the-boot ankle braces could reduce ankle sprains during Army paratrooper training. METHODS: The randomized trial involved 777 volunteers from the U.S. Army Airborne School, Fort Benning, Ga. Of this group, 745 completed all study requirements (369 brace-wearers and 376 non-brace-wearers). Each volunteer made five parachute jumps, for a total of 3,674 jumps. RESULTS: The incidence of inversion ankle sprains was 1.9% in non brace-wearers and 0.3% in brace-wearers (risk ratio, 6.9; p = 0.04). Other injuries appeared unaffected by the brace. Overall, 5.3% of the non-brace group and 4.6% of the brace group experienced at least one injury. The risk ratio for injured individuals was 1.2:1 (non-brace to brace groups; p = 0.65). CONCLUSION: Inversion ankle sprains during parachute training can be significantly reduced by using an outside-the-boot ankle brace, with no increase in risk for other injuries. PMID- 9751555 TI - Spanish version of the Burn-Specific Health Scale. AB - OBJECTIVE: To study the viability, reliability, and validity of the Spanish version of the Burn-Specific Health Scale. METHODS: The questionnaire was cross culturally adapted and translated and its psychometric properties were tested regarding their viability, reliability, and validity. A total of 115 patients discharged from the Burn Care Unit of the Alicante General Hospital were included in the study. RESULTS: One hundred fifteen patients were interviewed and completed a total of 156 questionnaires. Of these, 112 were self-administered (71.79%) by the patient, with an average completion time of 12 to 13 minutes (SD = 3.44 minutes). The test-retest reliability, internal consistency, criterion validity, and construct validity all proved satisfactory. CONCLUSION: The Spanish version of the Burn-Specific Health Scale is a reliable and valid instrument for use in the Spanish population, and its results are perfectly comparable with those obtained in the original English version. PMID- 9751556 TI - Preflight versus en route success and complications of rapid sequence intubation in an air medical service. AB - BACKGROUND: Maintenance of an airway in the air medically transported patient is of paramount importance. The purpose of this study is to compare preflight versus en route rapid sequence intubation (RSI)-assisted intubations and to determine the value of air medical use of RSI. METHODS: This study is a 31-month retrospective review of all patients intubated and transported by a large city air medical service. Subgroup analysis was based on whether patients were transported from a hospital or a scene and whether they were intubated preflight or en route. Information on age, Glasgow Coma Scale score, type of scene, ground time, and previous attempts at intubation was recorded. Complications included failures, multiple attempts at intubation, arrhythmias, and need for repeated paralytic agents. Comparisons were made using a confidence interval analysis. An alpha of 0.05 was considered significant; Bonferroni correction was used for multiple comparisons. RESULTS: Three hundred twenty-five patients were intubated and transported by Lifeflight during the study period. Two hundred eighty-eight patients were intubated using RSI (89%). The success rate was 97%. Preflight intubations were performed on 100 hospital calls and 86 scene calls. En route intubations were performed on 40 hospital cases and 62 scene calls. Patients who underwent preflight intubations were significantly younger than those who underwent en route intubations for both the hospital group (34 +/- 11 vs. 44 +/- 24 years, p < 0.05) and the scene group (27 +/- 13 vs. 32 +/- 16 years,p < 0.05). Otherwise, the demographic characteristics of the four groups were similar. Trauma accounted for 60 to 70% of hospital transfers and almost 95 to 100% of scene calls. Compared with preflight intubations, there was a significant decrease in ground time for hospital patients who were intubated en route (26 +/- 10 vs. 34 +/- 11 minutes, p < 0.05) and for scene patients who were intubated en route (11 +/- 8 vs. 18 +/- 9 minutes, p < 0.05). There were no significant differences between the groups for number of failures (9 of 288), arrhythmias (18 of 288), or necessity for repeated paralysis (8 of 288). Multiple intubation attempts were performed in more scene preflight patients (30 of 86, 35%) than scene en route patients (16 of 62, 26%), but this did not reach statistical significance. Even for patients having previous attempts at intubation, the success rate using RSI was 93% (62 of 67). CONCLUSION: Air medical intubations, both preflight and en route, for both scene calls and interhospital transports, can be done with a very high success rate. Rapid sequence intubation may improve the success rate. For scene calls, there was a significant decrease in ground time, and there was a trend toward fewer multiple intubation attempts when the patient was intubated en route instead of preflight. PMID- 9751557 TI - Controlled reopen suture technique for pyloric exclusion. AB - BACKGROUND: Pyloric exclusion had been widely used in the management of complicated duodenal injuries. The original concept of pyloric exclusion was that this technique would temporarily exclude the pylorus during the healing phase, but would subsequently allow resumption of normal gastrointestinal tract transit through the duodenum. The best method for pyloric exclusion has not been well established. Controversies exist regarding the need for a gastrojejunostomy and vagotomy as part of the procedure. None of these combinations can fulfill the original concept of pyloric exclusion and avoid late complications. METHODS: We developed a controlled reopen suture technique for pyloric exclusion. This technique was applied to nine patients (group II) with a complicated blunt duodenal injury over the past 5 years. The clinical courses and outcomes of these patients were compared with an eight-patient comparison group treated by pyloric exclusion and gastrojejunostomy (group I) over the same time period. RESULTS: All 17 patients survived. There were one early (duodenal wound leakage) and two late complications (marginal ulcers) in the group I patients. No delayed complications were found in the group II patients. The average hospital stay was about the same in both groups. CONCLUSION: The controlled reopen suture technique is a quick and simple procedure. In the treatment of a complicated blunt duodenal injury, if repair of the duodenal wound will not compromise the lumen, gastrojejunostomy and vagotomy can be omitted when using this technique. This technique offers the best combination of limited surgery in the severely injured patient, effective exclusion of the duodenum until after the healing has occurred, and allowance for the resumption of normal gastrointestinal tract transit through the duodenum. The late complications of gastrojejunostomy can also be avoided. PMID- 9751558 TI - Abdominal compartment syndrome. AB - The ACS is a clinical entity that develops from progressive, acute increases in IAP and affects multiple organ systems in a graded fashion because of differential susceptibilities. The gut is the organ most sensitive to IAH, and it develops evidence of end-organ damage before the development of the classic renal, pulmonary, and cardiovascular signs. Intracranial derangements with ACS are now well described. Treatment involves expedient decompression of the abdomen, without which the syndrome of end-organ damage and reduced oxygen delivery may lead to the development of multiple organ failure and, ultimately, death. Multiple trauma, massive hemorrhage, or protracted operation with massive volume resuscitation are the situations in which the ACS is most frequently encountered. Knowledge of the ACS, however, is also essential for the management of critically ill pediatric patients (especially those with AWD) and in understanding the limitations of laparoscopy. The role of IAH in the pathogenesis of NEC, central obesity co-morbidities, and pre-eclampsia/eclampsia remains to be fully studied. PMID- 9751559 TI - Posterior dislocation of the 11th thoracic vertebra: report of a pediatric case. PMID- 9751560 TI - Laparoscopic treatment of gastric and diaphragmatic injury produced by thoracoabdominal stab wound. PMID- 9751561 TI - Intussusception after blunt abdominal trauma. PMID- 9751563 TI - Gunshot wound traversing the ventricular septum with peripheral embolization presenting as complete heart block. PMID- 9751562 TI - Giant gastric ulcer in a body packer. PMID- 9751564 TI - Temporary pericardial replacement for thoracic damage-control procedures: avoidance of mechanical damage to the heart. PMID- 9751565 TI - Trauma-associated dissection of the thoracic aorta. PMID- 9751566 TI - Ligation of the suprarenal vena cava after a gunshot wound. PMID- 9751567 TI - Skeletal muscle pH, P(CO2), and P(O2) during resuscitation of severe hemorrhagic shock. PMID- 9751568 TI - Rupture of a traumatic common hepatic artery aneurysm: case report and review of the literature. PMID- 9751569 TI - Spleen rupture in the newborn: conservative surgical treatment using absorbable mesh. PMID- 9751570 TI - Laceration of the external carotid artery after an intraoral stab wound. PMID- 9751571 TI - Nitric oxide synthase as a therapeutic target in sepsis--more questions than answers? PMID- 9751572 TI - Nuclear factor-kappa B and nitric oxide regulating life and death: nonsense or harsh reality? PMID- 9751573 TI - Gastrointestinal motility and tube feeding. PMID- 9751574 TI - BIPAP: useful new modality or confusing acronym? PMID- 9751575 TI - Lactate and point-of-care testing. PMID- 9751576 TI - Optimal protein intake in critical illness? PMID- 9751577 TI - The importance of being selenium. PMID- 9751578 TI - Gastric mucosal pH is definitely obsolete--please tell us more about gastric mucosal PCO2. PMID- 9751579 TI - Endotoxin hyperventilation mechanisms. PMID- 9751580 TI - Monitoring cerebral oxygenation in the twilight years of the decade of the brain. PMID- 9751581 TI - Extracorporeal membrane oxygenation: are more descriptions needed? PMID- 9751582 TI - Extracorporeal membrane oxygenation and pediatric acute respiratory distress syndrome: we can afford it, but we don't need it. PMID- 9751583 TI - Institutional Review Board review and consent for research: what's behind the statistics? PMID- 9751584 TI - N(G)-monomethyl-L-arginine improves survival in a pig model of abdominal sepsis. AB - OBJECTIVE: To test the effect of a continuous infusion of the nitric oxide synthase inhibitor N(G)-monomethyl-L-arginine (L-NMMA) on survival rate and hemodynamics in a pig model of endogenous peritoneal live bacterial sepsis. DESIGN: Prospective, randomized trial. SETTING: Laboratory at a university medical center. SUBJECTS: Thirty-five pigs with an average weight of 26 kg (range 21 to 33). INTERVENTIONS: After surgical preparation, animals (control, n=6) given anesthesia and fluids were observed for 9 hrs. Fifteen experimental animals received 0.5 g of cecal content/kg of body weight intraperitoneally after surgery. Nine of these animals received standard anesthesia and fluids and were observed for 9 hrs or until death. Six animals received a continuous infusion of L-NMMA (10 mg/kg/hr) 3 hrs after sepsis induction. Starting 3 hrs after surgery, five nonrandomized animals were given anesthesia and fluids and received a 6-hr continuous infusion of L-NMMA (10 mg/kg/hr). An additional nine animals were anesthetized and blood samples were taken to determine plasma nitrate concentrations in nonoperated pigs. MEASUREMENTS AND MAIN RESULTS: L-NMMA treatment increased 9-hr survival in septic animals from 11% to 83% (p < .001), prevented a further decrease in mean arterial pressure and restored mean arterial pressure to control levels (p < .00002 vs. nontreated septic animals). Mean pulmonary arterial pressure increased slightly during L-NMMA infusion (p < .0003). Coronary blood flow was preserved during L-NMMA treatment. Cardiac index and urine production reached and maintained control levels during L-NMMA treatment of septic animals. Mean central venous pH did not deteriorate during L NMMA treatment. Animals treated with L-NMMA had plasma nitrate concentrations similar to nonseptic control animals. The results from the nonseptic control group receiving L-NMMA suggest that a substantial part of the effect of L-NMMA in this model of septic shock may be due to inhibition of the constitutive nitric oxide production. CONCLUSIONS: In this porcine model of peritoneal sepsis, infusion of L-NMMA increased survival rate and maintained mean arterial pressure without worsening tissue oxygenation. Coronary blood flow, cardiac index, systemic vascular resistance, and urine production were well maintained during L NMMA treatment. PMID- 9751585 TI - Nitric oxide inhibits stress-induced endothelial cell apoptosis. AB - OBJECTIVES: To determine a mechanism by which nitric oxide alters induction of stress-induced endothelial cell apoptosis in vitro. Apoptosis is a form of cellular suicide that has been implicated in the pathogenesis of multiple organ dysfunction syndrome. DESIGN: Prospective, controlled trial. SETTING: Research laboratory of a large, academic medical center. SUBJECTS: Cultured primary porcine aortic endothelial cells. INTERVENTIONS: Cells were treated with a range of doses of agents that either spontaneously generate nitric oxide (S-nitroso-N acetyl-D,L-penicillamine [SNAP] or (Z)-1-[2-(2-aminoethyl)-N-(2 ammonioethyl)amino]diazen-1- ium-1,2-diolate [DETA-NO]) or block nitric oxide production (Nomega-methyl-L-arginine [L-NMA]). The ability of these agents to alter the rate of cell death by apoptosis (induced by the sequence stimuli lipopolysaccharide [LPS] followed by sodium arsenite) was measured. Mechanistic studies included examining the ability of: a) nitric oxide "donors" to alter nuclear factor kappa B (NF-kappaB) DNA binding activity and the level of IkappaBalpha accumulation; and b) a stable cyclic guanosine monophosphate (cGMP) analog (8-bromo-cGMP) to mimic the effect of nitric oxide donors. MEASUREMENTS AND MAIN RESULTS: The sequence LPS/sodium arsenite increased the rate of endothelial cell apoptosis (47.4%, p< .05 vs. control), as measured by fluorescent-activated cell scanning using annexin V/propidium iodide staining. DETA-NO generated nitric oxide (as indicated by an increase in the concentration of the stable end-products of nitric oxide metabolism) and decreased the rate of endothelial cell apoptosis (20.6% at a dose of 2 mM, p=.0001 vs. control). DETA NO also decreased NF-kappaB DNA binding activity and the apparent accumulation of its endogenous inhibitor, IkappaBalpha. The 8-bromo-cGMP did not mimic the effects of nitric oxide donors (DETA-NO) on apoptosis. CONCLUSIONS: These data suggest that exogenous nitric oxide can block stress-induced endothelial cell apoptosis in vitro. The mechanistic studies are consistent with our hypothesis that inhibitors of NF-kappaB DNA binding activity are associated with protection against apoptosis-inducing stimuli. The results do not support a role for cGMP in mediating the protective effect of DETA-NO in our model. PMID- 9751586 TI - Gastrointestinal motility and gastric tube feeding in mechanically ventilated patients. AB - OBJECTIVE: To determine the fasted and fed gastrointestinal motility characteristics that are possibly responsible for gastric retention in mechanically ventilated patients. DESIGN: Prospective, case series. SETTING: Surgical intensive care unit of a university hospital. PATIENTS: Seven patients who required mechanical ventilation for thoracic or combined thoracic-neurologic injuries and nine healthy volunteers. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Antroduodenal manometry was performed during fasting and gastric feeding with a polymeric diet in patients during mechanical ventilation, weaning, and after detubation. Gastric retention volumes were determined during gastric tube feeding. Motility data were compared with recordings from nine healthy volunteers. During the fasting state, under sedation and morphine, the migrating motor complex in patients was significantly (p < .001) shortened: median 32.0 vs. 101.0 mins in healthy volunteers. During gastric tube feeding, the motility pattern did not convert to a normal postprandial pattern until morphine was discontinued. An interdigestive or mixed interdigestive-postprandial pattern was seen during gastric tube feeding in most patients during morphine administration. Most (94%) of the activity fronts during gastric feeding started in the duodenum. Gastric retention percentages during gastric tube feeding were negatively correlated (r2=.44; p < .01) with antral motor activity. CONCLUSIONS: These data suggest that morphine administration affects antroduodenal motility in mechanically ventilated patients. The gastrointestinal motor pattern involved in impaired gastric emptying in morphine-treated patients is characterized by antral hypomotility and persisting duodenal activity fronts during continuous intragastric feeding. The observed motility patterns suggest that early administration of enteral feeding might be more effective into the duodenum or jejunum than into the stomach of mechanically ventilated patients. PMID- 9751587 TI - Comparison of oxygen cost of breathing with pressure-support ventilation and biphasic intermittent positive airway pressure ventilation. AB - OBJECTIVE: To assess the oxygen cost of breathing with either pressure-support ventilation (PSV) or biphasic intermittent positive airway pressure ventilation (BIPAP). DESIGN: Prospective, randomized, crossover study. SETTING: Medical intensive care unit of a university hospital. PATIENTS: Twenty clinically stable and spontaneously breathing patients after long-term mechanical ventilation. INTERVENTIONS: Patients were randomized to start on either PSV or BIPAP, and measurements were performed after an adaptation period of 30 mins. Immediately after, the ventilatory mode was changed and after another 30-min adaptation period, the same measurements were performed. MEASUREMENTS AND MAIN RESULTS: Indirect calorimetry was performed during each ventilatory mode for a period of 30 mins. Oxygen consumption, energy expenditure, CO2 production, and respiratory quotient did not differ significantly between the two ventilatory modes, regardless of the patients' randomization. There were no statistically significant differences with regard to respiratory rate, minute volume, and blood gas analysis. All patients tolerated both ventilatory modes without any signs of discomfort. CONCLUSIONS: Pressure support ventilation and BIPAP are both used for weaning patients gradually from the ventilator. BIPAP may be advantageous in patients not breathing sufficiently with PSV, since no patient effort is necessary with use of this ventilatory mode. PMID- 9751588 TI - Validation of a hand-held lactate device in determination of blood lactate in critically injured patients. AB - OBJECTIVES: Admission blood lactate is an accurate predictor of injury severity and mortality in trauma patients. The purpose of this study was to evaluate a portable lactate analyzer in a clinical setting by patient care staff. DESIGN: A prospective, single-operator control solution and patient sample study, using two test devices and a reference device. SETTING: An urban Level I trauma center. PATIENTS: A convenience sample of 47 trauma patients. INTERVENTIONS: Intra-assay precision was demonstrated by performance of consecutive analyses of two lactate control solutions (high and low lactate control concentrations) by medical students and physicians. Split sample, simultaneous testing of the portable lactate analyzer was then performed on 66 whole blood specimens from a convenience sample of 47 trauma patients admitted to an urban Level 1 trauma center over 4 mos. Samples were tested simultaneously tested on two portable lactate analyzers and a reference instrument. MEASUREMENTS AND MAIN RESULTS: Acceptable intra-assay precision was achieved. Regression analysis for two test instruments demonstrated a slope of 0.920, an intercept of 0.323, an r2 of .982, and an SEM of 0.496. Regression analysis for test instrument "A" vs. the reference instrument showed a slope of 0.861, an intercept of 0.209, an r2 of .977, and an SEM of 0.598. Regression analysis for test instrument "B" vs. the reference instrument demonstrated a slope of 0.929, an intercept of -0.095, an r2 of .983, and an SEM of 0.506. CONCLUSIONS: Good correlation with a low SEM was obtained over a wide range of clinically relevant lactate values. Use of point of care lactate analysis will decrease analytic time, making an important diagnostic parameter immediately available in the critical care setting. PMID- 9751589 TI - Optimal protein requirements during the first 2 weeks after the onset of critical illness. AB - OBJECTIVE: To obtain optimal protein requirements in critically ill sepsis or trauma patients during the first 2 wks after admission to the intensive care unit. DESIGN: Retrospective study. SETTING: Department of critical care medicine at a teaching hospital. PATIENTS: Immediate posttrauma patients or severely septic patients. INTERVENTIONS: In vivo neutron activation analysis was used to measure changes in total body protein over a 10-day period which began as soon as the patients were hemodynamically stable. The patients (trauma, n=18; sepsis, n=5) were divided into three groups according to the average daily protein intakes. Because the patients were overhydrated (approximately 10 L) and had variable amounts of body fat, the protein intakes were indexed to normally hydrated (corrected) fat-free mass (FFMc): Groups A, B, and C received an average of 1.1, 1.5, and 1.9 g/kg FFMc/day protein, respectively. MEASUREMENTS AND MAIN RESULTS: Overall, the average loss of total body protein was 1.2=0.7 (SD) kg. Changes in total body protein were significantly (p=.011) different between the three groups. The loss of body protein was significantly more in group A compared with groups B (p=.013) and C (p=.023). When the protein intake was increased from 1.1 g/kg FFMc/day to 1.5 g/kg FFMc/day, protein loss was halved. Further increase in protein intake up to 1.9 g/kg FFMc/day resulted in no further improvement. An intake of 1.5 g/kg FFMc/day was equivalent to 1.0 g/day/kg of body weight measured at the beginning of the study. CONCLUSIONS: Current recommended protein requirements of 1.2 to 2.0 g/kg of body weight/day are excessive if they are indexed to the body weight measured soon after the onset of critical illness. Because individual patients have varying degrees of overhydration early in the illness onset, we suggest that the intensivist should obtain information on preillness body weight and prescribe 1.2g of protein/kg body weight/day. If information is not available, 1.0g of protein/day/kg of measured body weight will give a fair approximation to optimal protein requirements. PMID- 9751591 TI - Ketamine significantly reduces the migration of leukocytes through endothelial cell monolayers. AB - OBJECTIVE: To study the role of neutrophils in host defense, using human endothelial cells in migration studies in the presence of ketamine (0.3, 3, and 30 microg/mL). DESIGN: Prospective, controlled study. SETTING: University research laboratories. SUBJECTS: Seven independent experiments from different donors were done, investigating the influence of ketamine (0.3, 3, and 30 microg/mL) to the migration of human leukocytes through human endothelial cell monolayers. INTERVENTIONS: Human endothelial cell monolayers and/or human leukocytes were preincubated with clinically relevant (3 microg/mL), higher (30 microg/mL), and lower (0.3 microg/mL) concentrations of ketamine. The amount of leukocyte migration after 3 hrs was measured in a fluorometer. MEASUREMENTS AND MAIN RESULTS: Human endothelial cells isolated from umbilical veins were cultured on microporous membranes (polyethylene-terephthalat membranes) until they formed an endothelial cell monolayer. Leukocytes were separated by standard procedures. The migration of leukocytes through monolayers of endothelial cells under the clinically relevant concentration of ketamine was reduced to 59+/-9.8% (SD) (p< .05) when leukocytes but not the endothelial cell monolayers were preincubated with ketamine. Leukocyte migration was reduced to 92+/-7.3% (p > .05) when only monolayers of endothelial cells were treated with ketamine, and to 52+/-8.8% (p< .05) when both leukocytes and monolayers of endothelial cells were treated with ketamine. The higher and lower concentrations showed a dose-dependent effect. CONCLUSIONS: We investigated the cellular interaction between both cell systems, leukocytes and endothelial cells, simultaneously in the presence of ketamine. Ketamine is able to reduce significantly the migration of leukocytes through endothelial cell monolayers. The use of different dosages revealed a dose dependent effect. The current model allowed treatment of one cell type, either leukocyte or endothelial cell. Ketamine inhibits the function of leukocytes more than the function of endothelial cells. PMID- 9751590 TI - Selenium, systemic immune response syndrome, sepsis, and outcome in critically ill patients. AB - OBJECTIVES: To confirm early, marked decrease in plasma selenium concentrations in patients admitted to a surgical and medical intensive care unit (ICU), and to study this decrease according to the presence or absence of systemic inflammatory response syndrome (SIRS), sepsis, or direct ischemia-reperfusion. DESIGN: Prospective, observational study. SETTINGS: Collaboration between the adult ICU of a 1,100-bed general hospital and a biochemical research laboratory of a university medical center. PATIENTS: One hundred thirty-four consecutive surgical and medical ICU patients. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: In the first 31 patients, plasma and urine selenium concentrations were measured by electrothermal atomic absorption spectrometry on admission and once weekly during their ICU stay. These values were compared first with severity scores, criteria for SIRS, sepsis, and organ system failure taken on admission, and then with nosocomial infection, organ system failure during ICU stay, and hospital mortality. An early, low mean plasma selenium concentration was observed in these patients compared with selenium laboratory reference values. Plasma selenium, measured on ICU admission, inversely correlated with Acute Physiology and Chronic Health Evaluation II or Simplified Acute Physiology II scores. Patients with SIRS had lower selenium concentrations than those without SIRS. Mean urine selenium losses were normal in the first 31 patients. Plasma selenium concentration was low in all patients with severe sepsis and septic shock (range 0.20 to 0.72 micromol/L) and in those patients with ischemia-reperfusion from aortic cross clamping (range 0.34 to 0.68 micromol/L). Despite recommended specific selenium supplementation, plasma selenium concentrations remained low for >2 wks in patients with SIRS. However, there was a slight increase in plasma selenium concentrations in surviving SIRS patients, whereas plasma selenium concentrations decreased in nonsurviving patients. The frequency of ventilator-associated pneumonia, organ system failure, and mortality was three times higher in patients with low plasma selenium concentration at the time of admission (selenium < or =0.70 micromol/L) than for the other patients. CONCLUSIONS: In severely ill ICU patients with SIRS, we observed an early 40% decrease in plasma selenium concentrations, reaching values observed in deleterious nutritional selenium deficiency. This prolonged decrease in selenium concentrations could explain the three-fold increase in morbidity and mortality rates in these patients compared with other ICU patients. The efficacy of selenium treatment in SIRS patients with a high gravity index score or hypoperfusion needs further investigation. PMID- 9751592 TI - Continuous intramucosal PCO2 measurement allows the early detection of intestinal malperfusion. AB - OBJECTIVES: The intestinal metabolic and histologic changes that occur in the gastrointestinal tract with ischemia and that form the basis of intramucosal pH and PCO2 alterations have not been well established. Recent evidence suggests that apart from technical problems with gastric tonometry, some methodologic misconceptions in the interpretation of intramucosal pH and PCO2 exist. The present study was designed to demonstrate the effects of impaired mesenteric perfusion with specific consideration to the induced intramucosal PCO2 changes using a new technique, the continuous fiberoptic CO2 sensor, and a new concept of interpretation. DESIGN: Randomized, controlled intervention trial. SETTING: University animal laboratory. SUBJECTS: Twelve anesthetized female pigs, weighing 67+/-6 kg. INTERVENTIONS: The pigs were assigned to control and stenosis groups. In the stenosis group, blood flow in the superior mesenteric artery was reduced by 70% from baseline for 180 mins, followed by 120 mins of reperfusion. Serum lactate concentration, pH, PCO2, PO2, and bicarbonate concentration (cHCO3-) were determined in arterial, superior mesenteric venous, portal venous, hepatic venous, and pulmonary arterial blood. In the lumen of the ileum, intramucosal PCO2 was continuously determined by a fiberoptic CO2 sensor. At the end of the experiment, the gut was examined for histologic changes. MEASUREMENTS AND MAIN RESULTS: During mesenterial hypoperfusion, a sudden and significant increase in intramucosal PCO2 was observed. This increase was paralleled by increases in superior mesenteric venous PCO2 and portal venous PCO2 (p < .05) and a concomitant decrease in intramucosal pH, superior mesenteric venous pH, and portal venous pH. Arterial and mixed venous PCO2 and pH did not change. cHCO3- did not change in local or systemic blood samples. CONCLUSIONS: Compromised mesenteric blood flow causes significant metabolic and histologic changes. These local changes could not be detected by arterial or mixed venous lactate concentrations, pH, and PCO2 determinations. Under closed-system conditions, mesenteric CO2 accumulation causes an impairment of the CO2-HCO3- buffer, resulting in an unchanged cHCO3-. With impaired mesenteric perfusion, only intramucosal PCO2 alterations occur and an intramucosal pH calculation based on systemic cHCO3-changes is not necessarily correct. Therefore, the only parameter of importance is the intraluminal measurement of intramucosal PCO2 that can reflect isolated mesenteric changes. Thus, we recommended abolishing the terms "intramucosal pH measurement" and "gastric tonometry" and propose using the definition "intramucosal PCO2 measurement." PMID- 9751593 TI - Peripheral neural modulation of endotoxin-induced hyperventilation. AB - OBJECTIVE: To delineate the role of the peripheral neural reflexes involved in modulating hyperventilation during endotoxemia. DESIGN: A prospective, randomized, controlled, multigroup study. SETTING: Research animal laboratory. SUBJECTS: Adult Sprague-Dawley rats (n=43; 354+/-24 g) of either gender. INTERVENTIONS: Eight rats received a sham operation on their vagus, carotid sinus, and aortic nerves before the administration of a saline vehicle to serve as the time control. In the endotoxin group, 11 rats received a sham operation before endotoxin challenge. The remaining 24 rats received bilateral vagotomy (n=8), perivagal capsaicin treatment (n=8), or denervation of peripheral chemoreceptors (n=8) before endotoxin challenge. After the breathing pattern returned to a steady state, endotoxin (L-4130, serotype 0111, B4 lipopolysaccharide; 50 mg/kg) was injected into the vein. The rat's respiration was then monitored continuously for 5 hrs or until the animal died. MEASUREMENTS AND MAIN RESULTS: The respiratory rate and tidal volume did not change over the 5 hr observation period in the time control group. In the endotoxin group, the respiratory rate increased significantly from baseline (135.4%) 2 hrs after endotoxin challenge and increased persistently until the rats died. The tidal volume increased gradually to < or =132.8% of baseline 4 hrs after endotoxin challenge. Bilateral cervical vagotomy and perineural capsaicin treatment of the vagus nerves eliminated the tachypnea response to endotoxin injection. Denervation of the peripheral chemoreceptor accentuated the hyperventilation response to endotoxin, and resulted in the shortest survival time. CONCLUSIONS: Both lung vagal C-fiber afferents and peripheral chemoreceptors are involved in modulating the hyperventilation response after endotoxin challenge in rat models. Stimulation of vagal C-fiber afferents increased the respiratory rate. Conversely, the role of peripheral chemoreceptors was to restrain the hyperventilatory response and these receptors may play a protective role during endotoxemia. PMID- 9751594 TI - Oxygen uptake during peritoneal ventilation in a porcine model of hypoxemia. AB - OBJECTIVES: Peritoneal ventilation (PV) can greatly increase PaO2 in hypoxemic rabbits. We tested the hypothesis that the peritoneum can provide a gas exchange surface for oxygen uptake in larger animals that, like humans, have a smaller relative peritoneal surface area and corresponding blood flow. DESIGN: Prospective, randomized, controlled animal study. SETTING: University research laboratory. INTERVENTIONS: In six anesthetized pigs, a modified endotracheal tube (9.0-inner diameter) was inserted into the peritoneal cavity, and the peritoneal cavity was ventilated with oxygen in helium in gas phase. Measurements of peritoneal oxygen uptake and mixed venous oxygen saturation were made over 30 mins of: a) baseline FiO2 0.20, no PV; b) FiO2 0.20, PV; c) FiO2 0.20, PV, dopamine 5 microg/kg/min; d) baseline FiO2 0.15, no PV; e) FiO2 0.15, PV; and f) FiO2 0.15, PV, dopamine 5 microg/kg/min. MEASUREMENTS AND MAIN RESULTS: Mixed venous oxygen saturation was 61% at the baseline FiO2 of 0.20 and 33% at an FiO2 of 0.15 and did not increase significantly from baseline with PV or with dopamine at either FiO2. Peritoneal oxygen uptake, measured with a waterseal spirometer, was 9.1+/-3.1 (SD) and 11.9+/-3.0 mL/min when lung FiO2 was 0.20 and 0.15, respectively, and 9.7+/-2.8 and 12.2+/-2.7 mL/min when FiO2 was 0.20 and 0.15 and dopamine was infused, respectively. CONCLUSION: Peritoneal ventilation does not result in clinically significant oxygen uptake or alter mixed venous oxygen saturation in a porcine model of hypoxemia. PMID- 9751595 TI - Effects of catecholamines on the pulmonary venous bed in sheep. AB - OBJECTIVE: To assess the effects of various catecholaminergic agents on pulmonary venous tone. DESIGN: Prospective, randomized, controlled, experimental study. SETTING: Physiology laboratory of a university hospital. SUBJECTS: Thirty anesthetized, mechanically ventilated adult sheep. INTERVENTIONS: Four groups of six animals received 1-hr infusions of norepinephrine (0.5 microg/kg/min), epinephrine (0.5 microg/kg/ min), dopamine (10 microg/kg/min), or dobutamine (10 microg/kg/min). MEASUREMENTS AND MAIN RESULTS: A 7-Fr pulmonary artery catheter was placed in a proximal location to measure cardiac output and pressure in a large pulmonary vein (Ppw) after balloon inflation. Another catheter wedged in a small pulmonary artery measured pressure in a small pulmonary vein (Pdw). A third catheter measured left atrial pressure (PLA ). This method was able to detect the pulmonary venoconstrictive effects of histamine in a separate group of six animals. Pdw-PLA increased from a mean of 2.0+/-1.7 to 3.0+/-1.5 (SD) cm H2O (p < .01), 2.3+/-1.6 to 4.4+/-1.3 cm H2O (p < .01), and 1.7+/-1.0 to 3.5+/-2.2 cm H2O (p < .05) with norepinephrine, epinephrine, and dopamine, respectively. All of these drugs increased Pdw-Ppw, but only norepinephrine and epinephrine increased Ppw-PLA . No change in either pressure difference was observed with dobutamine. Elevation of cardiac output alone could not account for these findings since the increase in cardiac output induced by fluid infusion did not change the pressure differences. CONCLUSION: Norepinephrine, epinephrine, and dopamine at doses commonly used in humans increase pulmonary venous tone in sheep. PMID- 9751596 TI - Relationship of brain tissue PO2 to outcome after severe head injury. AB - OBJECTIVE: To determine thresholds of brain tissue PO2 (PbtO2) that are critical for survival after severe head injury. DESIGN: Prospective data collection. SETTING: Neurosurgical intensive care unit of Ben Taub General Hospital, a comprehensive academic neurosurgical facility and Level I trauma center. PATIENTS: Forty-three severely head-injured patients who were not obeying commands on presentation or whose condition deteriorated to this level shortly after admission. INTERVENTIONS: Intracerebral placement of Licox (n=39) or Paratrend (n=4) PO2 probes during craniotomy or in the intensive care unit. MEASUREMENTS AND MAIN RESULTS: PbtO2 monitoring continued for an average of 84.6+/-41.8 hrs. The probes were calibrated before insertion according to the manufacturer's specifications. After removal, probes were tested in room air and in blood gas standard calibration solutions. PbtO2 data were analyzed by comparing the average time that PbtO2 was below the values of 20, 15, 10, 8, 6, 4, and 2 torr (2.7, 2.0, 1.3, 1.0, 0.8, 0.5, and 0.3 kPa, respectively) in patients who were living 3 mos after injury vs. those who died. A Tobit regression analysis using maximum likelihood methods was utilized. Both Licox and Paratrend probes functioned well in room air and in the Level I control. However, in the zero-oxygen solution, the Paratrend probes gave an average reading of 7.0+/-1.4 torr (0.9+/-0.2 kPa), compared with 0.3+/-0.3 torr (0.04+/-0.04 kPa) for the Licox probes. CONCLUSIONS: Analysis of the PbtO2 monitoring data suggested that the likelihood of death increased with increasing duration of time at or below a PbtO2 of 15 torr (2.0 kPa) or with the occurrence of any PbtO2 values of < or =6 torr (< or =0.8 kPa). PMID- 9751597 TI - Cardiovascular complications adversely affect survival during extracorporeal membrane oxygenation. AB - OBJECTIVES: Extracorporeal membrane oxygenation (ECMO) has been used in the management of infants with cardiorespiratory failure. ECMO causes a decrease in load-dependent measures of cardiac performance that have not been demonstrated to affect patient outcome, while other cardiovascular complications occur which may affect outcome. The purpose of this study was to describe the cardiovascular complications associated with ECMO, and to determine their relationship to survival. DESIGN: Data were obtained, retrospectively, from the medical records of 500 consecutive newborns treated with ECMO at our institution since 1984. RESULTS: Hypertension (mean arterial pressure of >65 mm Hg) was the most common complication, requiring medical intervention in 192 infants. Myocardial stun, the near-total absence of systolic function during ECMO, occurred in 59 infants. Rhythm abnormalities, including noncannulation-related bradycardia, occurred in 43 infants, cardiac arrest, and pericardial effusion in 17 infants, and noninfective thrombosis in 9 infants. Only one infant required ligation of a patent ductus arteriosus during ECMO. Infants with at least one cardiovascular complication had a lower survival rate, compared with those infants without a complication. Survival rates were decreased in infants with myocardial stun, arrhythmia, and cardiac arrest. Hypertension and pericardial effusion were not associated with decreased survival. CONCLUSION: These findings suggest that some cardiovascular complications during ECMO are more common than previously thought, and cardiovascular complications may adversely impact outcome. PMID- 9751598 TI - Cost of extracorporeal life support in pediatric patients with acute respiratory failure. AB - OBJECTIVES: To determine the impact of extracorporeal life support (ECLS) on mortality in pediatric patients with acute hypoxemic respiratory failure (AHRF) at our institution; and to calculate the hospital charges associated with the use of ECLS. DESIGN: Retrospective review of medical records and hospital charges. SETTING: Pediatric intensive care unit (ICU) of a university-affiliated children's hospital. PATIENTS: Twenty patients admitted to the pediatric ICU between 1991 and 1995 for AHRF who received ECLS as a part of their hospital course. INTERVENTIONS: Predicted mortality was calculated using the Pediatric Respiratory Failure score and was compared with survival at the time of hospital discharge. Hospital charges were used as a proxy for resource utilization. Cost per-life-year-saved calculations were performed based on a normal life expectancy for survivors. MEASUREMENTS AND MAIN RESULTS: Twenty patients were identified. The median age was 4.83 yrs. The median duration of ECLS was 9 days, with 19.5 days in the pediatric ICU and 23.5 days for the entire hospital length of stay. The observed mortality rate for these patients was 20%. Median predicted mortality rate based on the Pediatric Respiratory Failure score calculation was 83%. The hospital charges incurred by these patients was a median of $199,096. Based on a normal life expectancy for survivors, this results in a cost of $4,190/life-year. CONCLUSIONS: ECLS for the pediatric patient with AHRF is done at a considerable cost. However, ECLS affects survival favorably, and compares favorably when considering cost/life-year calculations. The data presented in this study may serve as a benchmark for comparison with newer therapies (i.e., liquid ventilation, nitric oxide). These data also provide a framework for cost based analyses at other ECLS institutions. PMID- 9751599 TI - A report of the use of the Dynamic Objective Risk Assessment (DORA) score in the changing pediatric intensive care environment. AB - OBJECTIVE: To assess the clinical use of the Dynamic Objective Risk Assessment (DORA) severity of illness score in a site remote from its development. DESIGN: Prospective chart review. SETTING: Tertiary referral pediatric intensive care unit (PICU). PATIENTS: One hundred sixty consecutive admissions involving 621 patient days. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Pediatric Risk of Mortality (PRISM) scores were collected daily for all PICU patient days. Collection of data was performed by a physician not directly involved in the ordering of vital signs or laboratory data. The daily DORA score was calculated from the previous day's PRISM score and the admission PRISM score according to a previously described formula. The DORA score determines the patient's risk of mortality for the next 24 hrs. Also documented were the tests not ordered for each patient day. The sensitivity and specificity of the DORA score in our patient population were very similar to that previously reported using the previously described 1% cutoff for predicted mortality. We also noted that the tests ordered were related to the physician's perception of the patient's degree of sickness, and were themselves predictive of outcome. CONCLUSION: An outcome scoring system created in one group of PICUs can be applied to patients in another PICU remote from where the scoring system was developed with similar ability to predict outcome. PMID- 9751600 TI - Evaluation of Institutional Review Board review and informed consent in publications of human research in critical care medicine. AB - OBJECTIVE: To examine the frequency of obtaining Institutional Review Board (IRB) approval and informed consent in critical care research. DATA SOURCES AND DATA EXTRACTION: One-year retrospective review of original critical care research in humans published in seven journals, including American Journal of Respiratory and Critical Care Medicine, Chest, Critical Care Medicine, Intensive Care Medicine, The Journal of the American Medical Association, Lancet, and The New England Journal of Medicine. Studies were examined for general information (country/state where the research was performed, affiliation of the hospital to a medical school, and whether the work was supported by a grant and specifically by a pharmaceutical company), approval by IRB, method of consent, design of research, and interventions involved in the study. DATA SYNTHESIS: Two hundred seventy-nine studies were reviewed, 124 (44%) of which were conducted in the United States. Two hundred forty-three (87%) studies were performed in a university institution, 96 (34%) studies were supported by a grant, and 23 (24%) studies were supported by a pharmaceutical company. In 66 (24%) studies, there was no evidence of IRB review and informed consent approval. IRB approval was obtained but the method of consent was not specified in 36 (13%) studies. No significant differences were found in obtaining IRB approval and informed consent between research conducted in the United States (n=71, 57%) or outside the United States (n=92, 59%). Grant support was obtained in ten (9%) of the 116 studies not fully approved, compared with 70 (50%) of the 140 studies that obtained full approval (p < .05). All studies (23) supported by the pharmaceutical industry were fully approved. CONCLUSIONS: Many published studies in critical care lack IRB approval and/or informed consent. All research supported by the pharmaceutical industry was fully approved. The findings raise ethical concerns about critical care research. PMID- 9751601 TI - Understanding articles describing clinical prediction tools. Evidence Based Medicine in Critical Care Group. AB - OBJECTIVES: Clinical prediction rules and models are developed by applying statistical techniques to find combinations of predictors that categorize a heterogeneous group of patients into subgroups of risk. Our goal is to teach clinicians how to evaluate the validity, results, and applicability of articles describing clinical prediction tools. CLINICAL EXAMPLE: An article describing a rule to predict the need for intensive care unit care admission in patients presenting to the emergency room with chest pain. RECOMMENDATIONS: Valid clinical prediction tools are developed by completely following up a representative group of patients, by evaluating all potential predictors and testing the independent contribution of each predictor variable, and by ensuring that the outcomes were independent of the predictors. To evaluate the results of an article describing a clinical prediction tool, clinicians need to know what the prediction tool is, how well it categorizes patients into different levels of risk, and what the confidence intervals are around the risk estimates. Valid prediction tools are not applicable in every patient population. Before patient care application, the clinician should ensure that the tool maintains its prediction power in a new sample of patients, that the patients are similar to patients used to test the tool, and that the tool has been shown to improve clinical decision-making. CONCLUSIONS: There has been an increase in the development and validation of clinical prediction rules and models. It is important to evaluate the validity and reliability of these prediction tools before application. PMID- 9751602 TI - Beta-blocking agents can increase PaO2. PMID- 9751603 TI - Molecular epidemiology of human cancer: contribution of mutation spectra studies of tumor suppressor genes. PMID- 9751604 TI - Excess risk of colon cancer associated with a polymorphism of the APC gene? PMID- 9751605 TI - Somatic instability of the APC I1307K allele in colorectal neoplasia. AB - The adenomatous polyposis coli (APC) gene is proposed to function as a gatekeeper of colorectal neoplasia. A germ-line variant of this gene, the APC I1307K allele, is present in approximately 6% of the Ashkenazi Jewish population. To assess the role in tumorigenesis of the variant (A)8 tract produced by this allele, we undertook a somatic mutation analysis of the region surrounding codon 1307 in colorectal tumors from APC I1307K carriers. Somatic mutations involving the variant (A)8 tract were identified in 53 of 127 (42%) tumors from APC I1307K carriers compared with 5 of 127 (4%) mutations involving the wild-type allele of these tumors (P < 0.0001). Loss of heterozygosity of the wild-type allele was significantly more common in tumors with APC I1307K allele mutations (25 of 41, 61%) compared with APC I1307K carrier tumors without mutation of the variant (A)8 tract (12 of 53, 23%; P < 0.0005). This somatic biallelic APC inactivation further confirms the biological importance of the I1307K germ-line variant. The vast majority of APC I1307K somatic mutations consisted of a single adenine insertion (insA) involving the variant (A)8 tract. This insA mutation was mutually exclusive of the presence of microsatellite instability with 0 of 49 tumors with insA displaying BAT-26 instability compared with 9 of 78 tumors without insA (P=0.01). These findings support a model where somatic instability of the (A)8 tract produced by the APC I1307K allele leads to increased APC gene inactivation and directly accounts for 42% of the colorectal neoplasms occurring in APC I1307K carriers. PMID- 9751606 TI - Metallothionein deficiency promotes mouse skin carcinogenesis induced by 7,12 dimethylbenz[a]anthracene. AB - To investigate a possible involvement of metallothionein (MT) in chemical carcinogenesis, MT-I and MT-II gene-deficient [MT (-/-)] transgenic mice and wild type [MT (+/+)] control mice were topically applied at a single dose of 100, 250, 500, or 1000 microg of 7,12-dimethylbenz[a]anthracene (DMBA) on the dorsal skin and thereafter compared. After 14 weeks of DMBA treatment, the skin tumor occurred in MT (-/-) mice only and in a dose-dependent manner, whereas no change was observed in MT (+/+) mouse skin given the same DMBA treatment. The tumor cells showed proliferative activity, as shown by proliferative cell nuclear antigen staining. These results demonstrate that MT acts as an endogenous defensive factor against DMBA-induced skin tumorigenesis. PMID- 9751607 TI - Platelet-type 12-lipoxygenase in a human prostate carcinoma stimulates angiogenesis and tumor growth. AB - Previously, we found a positive correlation between the expression of platelet type 12-lipoxygenase (12-LOX) and the progression of human prostate adenocarcinoma (PCa; Gao et al., Urology, 46: 227-237, 1995). To determine the role of 12-LOX in PCa progression, we generated stable 12-LOX-transfected PC3 cells, which synthesize high levels of 12-LOX protein and 12(S) hydroxyeicosatetraenoic acid metabolite. In vitro, 12-LOX-transfected PC3 cells demonstrated a proliferation rate similar to neo controls. However, following s.c. injection into athymic nude mice, 12-LOX-transfected PC3 cells formed larger tumors than did the controls. Decreased necrosis and increased vascularization were observed in the tumors from 12-LOX-transfected PC3 cells. Both endothelial cell migration and Matrigel implantation assays indicate that 12-LOX-transfected PC3 cells were more angiogenic than their neo controls. These data indicate that 12-LOX stimulates human PCa tumor growth by a novel angiogenic mechanism. PMID- 9751608 TI - Close correlation between telomerase expression and adenomatous polyp progression in multistep colorectal carcinogenesis. AB - To investigate the role of telomerase in the multistep colorectal carcinogenesis, we examined telomerase activity in 31 adenomatous polyps and 22 paired cancer normal mucosa specimens from non-hereditary nonpolyposis colorectal cancer patients. Telomerase activity was detected in 18% of normal mucosa, 16% of small (<1.0 cm) polyps, 20% of intermediate polyps, 71% of large (>2.0 cm) polyps, and 96% of adenocarcinoma samples (P for trend, <0.0001). High-level enzyme activities were seen in none of the normal mucosa, 5% of small polyps, 20% of intermediate polyps, 43% of large polyps, and 73% of adenocarcinoma samples (P for trend, <0.0001). These data indicate telomerase reactivation occurs with adenomatous polyp progression in multistep colorectal carcinogenesis. PMID- 9751609 TI - Constitutive and antibody-induced internalization of prostate-specific membrane antigen. AB - Prostate-specific membrane antigen (PSMA) is a cell surface glycoprotein expressed predominantly by prostate cancer cells. We have characterized four monoclonal antibodies that bind to the extracellular domain of PSMA (Liu et al., Cancer Res., 57: 3629-3634, 1997). Here we report that viable LNCaP cells internalize these antibodies. Laser scanning confocal microscopy reveals that the internalized antibodies accumulate in endosomes, and immunoelectron microscopy reveals that endocytosis of the PSMA-antibody complex occurs via clathrin-coated pits. In addition, a quantitative cell surface biotinylation assay demonstrates that PSMA is constitutively endocytosed in LNCaP cells and that anti-PSMA antibodies increase the rate of internalization of PSMA. These studies suggest that PSMA might function as a receptor mediating the internalization of a putative ligand. The availability of prostate-specific internalizing antibodies should aid the development of novel therapeutic methods to target the delivery of toxins, drugs, or short-range isotopes specifically to the interior of prostate cancer cells. PMID- 9751610 TI - Metastatic cutaneous melanoma promoted by ultraviolet radiation in mice with transgene-initiated low melanoma susceptibility. AB - An inbred-strain (C57BL/6) transgenic (Tyr-SV40E) mouse model of ultraviolet radiation (UVR)-induced metastatic cutaneous melanoma was produced without the use of chemical carcinogens and without resulting in other skin malignancies. Expression of this transgene occurs specifically in melanocytic-lineage cells. In untreated hemizygous mice of transgenic line 12 there are no skin melanomas, and the oncogenic sequence, which is expressed at a very low level, functions solely as a weak initiating stimulus. UVR [including 65% ultraviolet B (280-320 nm wavelength)] supplied the necessary promoting stimulus leading to melanomas. Of various trial protocols, eight were successful and involved exposure of 112 mice for a limited time on each of 3-10 days starting at 2-3 days of age and totalling 1.1-3.7 J/cm2 UVR. Fourteen of these animals developed a total of 15 invasive skin melanomas on the head and body, arising between 37-115 weeks of age and, therefore, often after a relatively long latency. The tumors were melanotic and in five of the mice they yielded macrometastases in regional and distant sites. The single most favorable protocol (1.9 J/cm2 total UVR, at 0.38 J/cm2/day for 5 days starting at 3 days of age) led to the highest incidence of melanoma (5 of 19 mice) and one of the lowest mortality rates (2 of 19). No melanomas occurred in UVR-treated nontransgenic C57BL/6 controls. Benign skin keratoacanthomas arose and often regressed in treated transgenic as well as nontransgenic mice. This new transgenic mouse model introduces many novel possibilities for experimental analysis of the melanoma-promoting mechanisms of UVR and also of the ability of specific genetic changes to impede or facilitate the UVR effect. PMID- 9751611 TI - Multiplex reverse transcription polymerase chain reaction assessment of sialyltransferase expression in human breast cancer. AB - Increased sialylation, especially involving the Sialyl-Lewisa and Sialyl-Lewisx determinants, has been reported in breast cancer. A multiplex reverse transcription-PCR method was used here to determine the expression of five sialyltransferases (ST3Gal III, ST6Gal I, ST3Gal IV, ST3Gal I, and ST3Gal II) in 49 patients surgically treated for locoregional breast cancer. We assessed the relationship between these expressions and clinical, pathological, and biological features. The most expressed sialyltransferase was ST3Gal 1II, which is involved in Sialyl-Lewisa synthesis. ST3Gal III expression was positively correlated to ST6Gal I and ST3Gal IV expressions, to tumor size, and to the number of involved axillary nodes. Patients with high ST3Gal III expression had a shorter overall survival. High ST6Gal I expression was associated with histoprognostic grade III. ST6Gal I expression was negatively correlated to expression of progesterone receptor. In conclusion, high ST3Gal III and ST6Gal I expressions in human breast tumors are associated with poor prognosis markers. PMID- 9751612 TI - Elevated mitogen-activated protein kinase activity in estrogen-nonresponsive human breast cancer cells. AB - The mitogen-activated protein kinase (MAPK) signal transduction pathway plays an essential role in cell cycle progression and can be activated by many growth factor/mitogen pathways including estrogen. MAPK has also been implicated in ligand-independent activation of estrogen receptor-alpha (ER-alpha). The development of estrogen-independent growth in breast cancer is likely a first step in progression to hormone independence and antiestrogen resistance. We examined MAPK expression and activity in T5-PRF and T5 human breast cancer cells. T5-PRF is an estrogen-nonresponsive cell line developed from T5 cells by chronically depleting the cells of estrogen in long-term culture. MAPK activity measured in vitro was significantly higher (P < 0.05) in T5-PRF compared with T5 cells. Western blot analyses showed increased levels of active dually phosphorylated MAPK in T5-PRF cell extracts compared with T5. The increased activity and expression of MAPK may contribute to the estrogen nonresponsive growth phenotype and ligand-independent activity of ER in T5-PRF cells. PMID- 9751613 TI - Intratumoral conversion of 5-fluorocytosine to 5-fluorouracil by monoclonal antibody-cytosine deaminase conjugates: noninvasive detection of prodrug activation by magnetic resonance spectroscopy and spectroscopic imaging. AB - The monitoring of antibody-directed enzyme-prodrug therapies requires evaluation of drug activation within the tissues of interest. We have demonstrated the feasibility of noninvasive magnetic resonance spectroscopy and spectroscopic imaging (chemical shift imaging) to detect activation of the prodrug 5 fluorocytosine (5-FCyt) to the cytotoxic species 5-fluorouracil (5-FU) by monoclonal antibody-cytosine deaminase (CD) conjugates. In vitro, L6-CD but not 1F5-CD selectively metabolized 5-FCyt to 5-FU on H2981 human lung adenocarcinoma cells because of the presence and absence of cell surface L6 and CD20 antigens, respectively. After pretreatment of H2981 tumor-bearing mice with L6-CD, in vivo metabolism of 5-FCyt to 5-FU within the tumors was detected by 19F magnetic resonance spectroscopy; the chemical shift separation between 5-FCyt and 5-FU resonances was approximately 1.2 ppm. 5-FU levels were 50-100% of 5-FCyt levels in tumors 10-60 min after 5-FCyt administration. Whole body 19F chemical shift imaging (6 x 6 mm in-plane resolution) of tumor-bearing mice demonstrated the highest signal intensity of 5-FU within the tumor region. This study supports further development of noninvasive magnetic resonance methods for preclinical and clinical monitoring of CD enzyme-prodrug therapies. PMID- 9751614 TI - Expression of MUC1 mucin on activated human T cells: implications for a role of MUC1 in normal immune regulation. AB - MUC1 mucin is expressed by normal and malignant epithelial cells and is thought to function through cell-cell interactions and transmembrane signal transduction events. Secreted cancer-associated MUC1 is immunosuppressive and inhibits human T cell proliferation. We report here that newly synthesized MUC1 is expressed on the surface of mitogen-activated human T cells and is also found in soluble form in the supernatants from cultures of mitogen-activated human T cells. After removal of the mitogenic stimulus from the T-cell cultures, MUC1 expression is downregulated. The addition of anti-MUC1 monoclonal antibody to mitogen-activated cultures partially inhibits the T-cell proliferative response. These data suggest that MUC1 serves an immunodulatory function for human T lymphocytes. PMID- 9751615 TI - Overexpression of cdc25A and cdc25B is frequent in primary non-small cell lung cancer but is not associated with overexpression of c-myc. AB - Cyclin-dependent kinases can be activated by cdc25, which removes inhibitory phosphates from tyrosine and threonine residues. At least three cdc25 genes (cdc25A, cdc25B, and cdc25C) have been identified in humans. Accumulating evidence indicates that cdc25A and cdc25B possess oncogenic properties. Recently, overexpression of cdc25A and of cdc25B was found in many breast and head and neck cancers. To determine potential roles of cdc25s in non-small cell lung cancer (NSCLC), we analyzed primary tumors and corresponding normal lung tissues from 40 patients with NSCLC for relative expression levels of these genes by multiplex reverse transcription PCR (RT-PCR). cdc25A was overexpressed in 60% (24 of 40) of the tumors and cdc25B in 45% (18 of 40) of the tumors, whereas cdc25C was not overexpressed in any of the tumors analyzed. Because c-myc can increase cdc25A and cdc25B expression, it may be a factor in cdc25 overexpression. We found that c-myc was overexpressed in only 18% (7 of 40) of the tumors. We found no association between overexpression of c-myc and cdc25A or cdc25B. We also investigated whether the cdc25B gene was amplified in NSCLC and found this was true in 40% (8 of 20) of the tumors tested. However, this amplification was not correlated with gene expression status. Interestingly, among 24 tumors with cdc25A overexpression and 18 with cdc25B overexpression, 42% (10 of 24) and 44% (8 of 18) were poorly differentiated histological type. In contrast, well or moderately differentiated tumors had lower frequencies of cdc25A and cdc25B overexpression [19% (3 of 16) and 23% (5 of 22), respectively]. These data indicate that overexpression of cdc25A and cdc25B is frequent and that it may play an important role in NSCLC. However, it is unlikely that this overexpression is caused by c-myc stimulation or cdc25B gene amplification. PMID- 9751616 TI - Identification of germ-line E-cadherin mutations in gastric cancer families of European origin. AB - E-cadherin germ-line mutations have recently been described as a molecular basis for early-onset familial gastric cancer in Maori kindred. We screened 18 gastric cancer families of European origin for germ-line mutations to determine the proportion in which E-cadherin mutations occur and the clinical characteristics of the affected families. Truncating mutations were identified in three kindred with familial diffuse gastric cancer. In these families, the age of onset of gastric cancer was variable, the penetrance was incomplete, and one kindred contained individuals with cancers at other sites. Here, we show that a proportion of diffuse gastric cancer families of European origin have germ-line E cadherin mutations; however, these mutations are absent in intestinal gastric cancer families. PMID- 9751617 TI - Heterogeneous expression of the tumor-associated antigens RAGE-1, PRAME, and glycoprotein 75 in human renal cell carcinoma: candidates for T-cell-based immunotherapies? AB - It has recently been shown that tumor-associated antigens (TAAs) can evoke tumor specific T-cell-defined immune responses in cancer patients, thereby offering the possibility of treating patients with such antigens. To develop T-cell-based immunotherapeutic approaches for renal cell carcinoma (RCC), we studied the mRNA expression profile of the TAAs RAGE-1, tyrosinase, MAGE-1, MAGE-2, NY-ESO-1, Melan-A/MART-1, glycoprotein (gp) 75, gp100, beta-catenin, PRAME, and MUM-1 in 14 human RCC cell lines and in tissue specimens of 37 primary RCCs, 2 related metastases, and 33 specimens of normal renal epithelium. Reverse transcription PCR was performed with TAA-reactive primers, and the specificity of the PCR products was confirmed by Southern blot and/or direct sequencing. PRAME (10 of 14 cell lines), RAGE-1 (7 of 14 cell lines), and gp75 (4 of 14 cell lines) antigens were expressed in a high percentage of RCC cell lines, although the level of TAA expression varied among the different RCC cell lines. However, low levels of TAA expression in RCC cells are sufficient for recognition by TAA-specific CTLs. Transcription of tyrosinase, Melan-A/MART-1, MAGE-1, MAGE-2, NY-ESO-1, gp100, beta-catenin, and MUM-1 was not detected in any RCC cell line. Approximately 50% of surgically removed neoplasias expressed at least one TAA. RAGE-1 mRNA expression was found in 8 of 39 (21%) RCC samples, PRAME mRNA expression was found in 15 of 39 (40%) RCC samples, and gp75 mRNA expression was found in 4 of 39 (11%) RCC samples, but the expression levels of these TAAs were heterogeneous in the different RCC lesions. One RCC specimen expressed MAGE-2, whereas transcription was not detected in any RCC specimen for MAGE-1, NY-ESO-1, tyrosinase, Melan-A/MART-1, gp100, beta-catenin, and MUM-1. The normal kidney epithelium samples were negative for any TAA tested. Thus, RAGE-1, PRAME, and gp75 expression is found with a different frequency in surgically removed lesions and in RCC cell lines, suggesting that a subgroup of RCC patients could be selected for immunotherapeutic strategies that may benefit from immunization against the RAGE-1, gp75, and/or PRAME antigens. However, additional targets for T-cell-based immunotherapy of RCC have yet to be identified. PMID- 9751618 TI - Inhibition of lung carcinogenesis by black tea in Fischer rats treated with a tobacco-specific carcinogen: caffeine as an important constituent. AB - Here, we examined the effect of black tea and caffeine on lung tumorigenesis in F344 rats induced by the nicotine-derived carcinogen 4-(methylnitrosamino)-1-(3 pyridyl)-1-butanone (NNK) in a 2-year bioassay. NNK was administered s.c. at a dose of 1.5 mg/kg body weight three times weekly for 20 weeks. Animals were given either black tea as drinking water at concentrations of 2%, 1%, or 0.5%, or caffeine in drinking water at concentrations identical to those in 2% and 0.5% tea infusions for 22 weeks. The treatment period began 1 week before and ended 1 week after the NNK administration. The animals were sacrificed on week 101 for the examination of tumors in target organs, including lung, liver, nasal cavity, and other major organs. The NNK-treated group, given 2% black tea, showed a significant reduction of the total lung tumor (adenomas, adenocarcinomas, and adenosquamous carcinomas) incidence from 47% to 19%, whereas the group given 1% and 0.5% black tea showed no change. The 2% tea also reduced liver tumor incidence induced by NNK from 34% in the group given only deionized water to 12%. The tumor incidence in the nasal cavity, however, was not affected by either black tea or caffeine at any of the concentrations tested. The most unexpected finding was the remarkable reduction of the lung tumor incidence, from 47% to 10%, in the group treated with 680 ppm caffeine, a concentration equivalent to that found in the 2% tea. This incidence is comparable to background levels seen in the control group. This study demonstrated for the first time in a 2-year lifetime bioassay that black tea protects against lung tumorigenesis in F344 rats, and this effect appears to be attributed, to a significant extent, to caffeine as an active ingredient of tea. PMID- 9751619 TI - Essential role of p53 in phenethyl isothiocyanate-induced apoptosis. AB - Phenethyl isothiocyanate (PEITC) is a natural product that is among the most effective cancer chemopreventive agents known. Mechanistic studies indicate that the chemopreventive activity of PEITC is associated with its favorable modification of carcinogen metabolism and its induction of apoptosis. Here, we found that PEITC blocks tumor promoter (12-O-tetradecanoylphorbol-13-acetate or epidermal growth factor)-induced cell transformation in mouse epidermal JB6 cells, and this inhibitory activity on cell transformation is correlated with induction of apoptosis. Most importantly, apoptosis induction by PEITC occurs through a p53-dependent pathway. This was demonstrated not only by results that PEITC induction of p53 protein expression and p53-dependent transactivation but also by PEITC-induced apoptosis in p53 +/+ cells but not in p53 -/- cells. In contrast, PEITC induced apoptosis in cells with both normal or deficient sphingomyelinase activity. Our results demonstrate for the first time that p53 elevation is required for PEITC-induced apoptosis, which may be involved in its cancer chemopreventive activity. PMID- 9751620 TI - Involvement of dendritic cell response to resistance of stomach carcinogenesis caused by N-methyl-N'-nitro-N-nitrosoguanidine in rats. AB - The involvement of immune response in the resistance of chemically induced stomach cancer was studied in a resistant rat strain (Buffalo) and a sensitive rat strain (ACI). Groups of 10 male Buffalo and ACI rats, 6 weeks of age, were given drinking water with or without N-methyl-N'-nitro-N-nitrosoguanidine (MNNG; 100 mg/l) for 14 days. Total RNA was isolated from the stomach pyloric mucosa from five rats, and cDNA was prepared with reverse transcriptase. Tissue sections of the stomach pyloric mucosa from five rats were stained with antibodies recognizing molecules expressed by various immune cells. Reverse transcription PCR (RT-PCR), competitive RT-PCR, and Northern blot demonstrated that the expression of MHC class II group genes [MHC class II, MHC class II-associated invariant chain (Ii), CD4 and IgM (B cell marker)], MHC class I group genes (MHC class I and CD8), B7-1 (costimulator on dendritic cells), and CD28 (receptor to B7 on T cells) in the pyloric mucosa was elevated by MNNG in both rat strains but was elevated to a 4-7-fold greater extent in Buffalo rats than in ACI rats. These genes were scarcely expressed in control rats. Histochemical antibody staining after MNNG exposure showed a greater number of cells stained with monoclonal antibody to Ii, OX-62 (dendritic cell marker), and ED-1 (dendritic cell and macrophage common marker) in the interstitial tissue of the pyloric mucosa of Buffalo rats compared with ACI rats. Cell proliferation, as measured by 5-bromo-2 deoxyuridine (BrdUrd)-labeling indices, revealed the presence of BrdUrd-labeled cells only among epithelial cells in the proliferative zone; cells in the interstitial tissue were not labeled with BrdUrd. The results suggest the involvement of dendritic cell response in the resistance to the MNNG induction of stomach carcinogenesis in rats. PMID- 9751621 TI - Parathyroid hormone-related protein in patients with primary breast cancer and eucalcemia. AB - Parathyroid hormone-related protein (PTHrP) is a causative factor of humoral hypercalcemia in breast cancer and other malignancies. We studied circulating PTHrP levels with three different immunoassays directed against different parts of the PTHrP molecule in 48 patients with breast cancer and eucalcemia. The methods used were: (a) a RIA with antibodies directed toward the midregion (63 78); (b) an immunofluorometric assay with two antibodies against 1-34 and 38-67; and (c) an immunoradiometric assay with antibodies against 1-40 and 1-72. Although most patients had PTHrP levels indistinguishable from normal when measured by all three methods, four patients had increased serum levels in the IFMA. PTHrP was detected by immunohistochemistry in tumors from nearly all patients. One patient with elevated PTHrP in plasma measured by IFMA showed intense staining of tumor by immunohistochemistry; the tumor was histologically graded as III (severe) and was the largest of all tumors in this patient group. The IFMA can identify increased serum PTHrP in some patients with breast cancer who are not hypercalcemic. This assay may be especially useful in screening patients for this tumor during a relative early phase of the disease. PMID- 9751622 TI - Chromosomal aberrations in lymphocytes predict human cancer: a report from the European Study Group on Cytogenetic Biomarkers and Health (ESCH). AB - Chromosomal aberrations (CAs), sister chromatid exchanges (SCEs), and micronuclei (MN) in peripheral blood lymphocytes have for decades been used as cytogenetic biomarkers to survey genotoxic risks in the work environment. The conceptual basis for this application has been the idea that increased cytogenetic damage reflects an enhanced cancer risk. Nordic and Italian cohorts have been established to evaluate this hypothesis, and analyses presented previously have shown a positive trend between CA frequency and increased cancer risk. We now report on a pooled analysis of updated data for 3541 subjects examined for CAs, 2703 for SCEs, and 1496 for MN. To standardize for interlaboratory variation, the results for the various cytogenetic end points were trichotomized on the basis of the absolute value distribution within each laboratory as "low" (1-33 percentile), "medium" (34-66 percentile), or "high" (67-100 percentile). In the Nordic cohort, there was an elevated standardized incidence ratio (SMR) for all cancer among subjects with high CA frequency [1.53; 95% confidence interval (CI), 1.13-2.05] but not for those with medium or low CA frequency. In the Italian cohort, a SMR in cancer of 2.01 (95% CI, 1.35-2.89) was obtained for those with a high CA frequency level, whereas the SMRs for those with medium or low did not noticeably differ from unity. Cox's proportional hazards models gave no evidence that the effect of CAs on total cancer incidence/mortality was modified by gender, age at test, or time since test. No association was seen between the SCEs or the MN frequencies and subsequent cancer incidence/mortality. The present study further supports our previous observation on the cancer predictivity of the CA biomarker, which seems to be independent of age at test, gender, and time since test. The risk patterns were similar within each national cohort. This result suggests that the frequency of CAs in peripheral blood lymphocytes is a relevant biomarker for cancer risk in humans, reflecting either early biological effects of genotoxic carcinogens or individual cancer susceptibility. PMID- 9751623 TI - Serum antibodies to benzo(a)pyrene diol epoxide-DNA adducts in the general population: effects of air pollution, tobacco smoking, and family history of lung diseases. AB - Polycyclic aromatic hydrocarbons (PAHs) are almost ubiquitous pollutants that may interact with metabolic systems in human tissues and eventually cause cancer. PAH adducted DNA becomes antigenic and antibodies anti-benzo(a)pyrene diol epoxide (BPDE)-DNA may be found in serum of PAH-exposed subjects. The presence of serum antibodies anti-BPDE-DNA adduct was investigated in 1345 individuals from family clusters of the general population of a small area in central Italy in whom information about smoking habits, site of residence, and personal and family history of lung diseases, including cancer, were obtained. Anti-BPDE-DNA antibodies in the sera were detected with a direct ELISA and the association of anti-BPDE-DNA antibodies with subjects' data from a standardized respiratory questionnaire including age, occupation, tobacco smoking habits, respiratory symptoms, and family history of respiratory diseases was subsequently tested by multivariate logistic regression analysis. The overall prevalence of subjects with anti-BPDE-DNA antibodies was 21.0% (n=283), with no differences between males and females. Anti-BPDE-DNA positivity was associated with living in the urban area [odds ratio (OR), 1.49; 95% confidence interval (CI), 1.16-1.92], with active tobacco smoking (OR, 1.25; 95% CI, 1.06-1.48), and with family history of lung cancer (OR, 1.30; 95% CI, 0.90-1.88), and positivity increased with the number of members in the family cluster positive to anti-BPDE-DNA antibodies (OR, 1.30; 95% CI, 1.03-1.65). This study on a large general population sample indicates that serum anti-BPDE-DNA antibodies may be considered as biomarkers of exposure to environmental carcinogens and of DNA damage. The genetic and familial components of their association with tobacco smoking lend further support to the argument about the familial predisposition to lung cancer. PMID- 9751624 TI - Blockage of insulin-like growth factor-I receptor inhibits the growth of Ewing's sarcoma in athymic mice. AB - Innovative, more effective treatment modalities are needed for Ewing's sarcoma (ES), a neoplasm with a disappointingly low survival rate despite the use of aggressive multimodal therapeutic approaches. We have previously shown (K. Scotlandi et al, Cancer Res., 56: 4570-4574, 1996) the existence and the pathogenetic relevance of an autocrine loop that is mediated by the insulin-like growth factor-I receptor (IGF-IR) and is crucial for the survival and proliferation of ES cells in vitro. In this study, we report that the IGF-IR blocking monoclonal antibody alphaIR3 may also significantly inhibit ES cell growth in vivo. In particular, in almost one-half of the animals tested, after s.c. inoculation with TC-71 ES cells, the blockage of IGF-IR by alphaIR3 induced a complete regression of tumors that developed, which suggests that IGF-IR is valuable as a specific target for novel therapeutic strategies. In addition, suramin, a drug that can interfere with growth factor binding with their receptors, inhibited the tumorigenic and the metastatic ability of TC-71 cells and, therefore, is a promising agent to be combined with conventional cytotoxic drugs for the design of more effective therapeutic regimens. PMID- 9751625 TI - Targeted cytotoxic analogue of somatostatin AN-238 inhibits growth of androgen independent Dunning R-3327-AT-1 prostate cancer in rats at nontoxic doses. AB - Receptors for somatostatin (SST) that are found on prostate cancers might be used for targeting of chemotherapeutic agents. Thus, doxorubicin derivative 2 pyrrolinodoxorubicin (AN-201) can be linked to SST analogue RC-121 (D-Phe-Cys-Tyr D-Trp-Lys-Val-Cys-Thr-NH2) to form targeted cytotoxic SST analogue AN-238. In this study, we evaluated the effects of AN-238 on the growth of SST receptor (SSTR)-positive androgen-independent Dunning R-3327-AT-1 prostate cancers in Copenhagen rats. The dose range and tumor growth-inhibitory effects of AN-238 and AN-201 were investigated in preliminary experiments. Administration of cytotoxic radical AN-201 at single i.v. doses of 110, 125, and 150 nmol/kg resulted in 0, 77.7, and 100% mortality, respectively, within 6-10 days. Four weeks after the injection of 110 nmol/kg AN-201, mean tumor volume was reduced by 35.1 % (P < 0.05), as compared with controls. In contrast, a single i.v. injection of analogue AN-238 at a dose of 300 nmol/kg was nontoxic and remarkably potent in inhibiting the growth of Dunning AT-1 tumors, resulting in a 85.9% (P < 0.01) reduction in tumor volume after 4 weeks. Treatment with AN-238 extended the survival time of tumor-bearing rats from 52.0+/-3.75 to 91.8+/-3.70 days, corresponding to a 76.5% (P < 0.01) increase. In a comprehensive experiment, we compared the effects of radical AN-201 at 115 nmol/kg, analogue AN-238 at 115 and 300 nmol/kg, carrier SST analogue RC-121 at 300 nmol/kg, and a mixture of AN-201 and RC-121 at doses of 300 nmol/kg administered i.v. Administration of AN-201 at 115 nmol/kg led to 90.0% mortality in 12 days, but animals treated with 115 nmol/kg of AN-238 showed no signs of toxicity, their tumor volume was reduced by 40.0% (P < 0.05), and their tumor weight was reduced by 42.8% (P < 0.01) after 4 weeks, as compared with controls. The dose of 300 nmol/kg of AN-238 was also nontoxic and diminished tumor volume by 80.9% (P < 0.01) and tumor weight by 82.0% (P < 0.01). No reduction in tumor growth or toxic effects was observed with carrier RC-121, but after the injection of unconjugated mixture of AN-201 and RC 121 at doses of 300 nmol/kg, all rats died within 4 days. Specific high-affinity receptors for SST were found on Dunning R-3327-AT-1 tumor membranes by radioligand binding assay and were identified by reverse transcription-PCR as SSTR2. Our study indicates that cytotoxic SST analogue AN-238 can be targeted to SSTRs on tumors and produces a powerful inhibition of the growth of Dunning-AT-1 rat prostate cancer at doses that are nontoxic, whereas its cytotoxic component, 2-pyrrolinodoxorubicin, is toxic and ineffective. PMID- 9751626 TI - Constitutive up-regulation of integrin-mediated adhesion of tumor-infiltrating lymphocytes to osteoblasts and bone marrow-derived stromal cells. AB - Tumor-reactive T cells, known as tumor-infiltrating lymphocyte(TIL)s are known to infiltrate various tumors. Although TILs exert cytotoxic activities against tumor cells, only a small percentage of tumors usually contain TILs that specifically react to tumor antigens. Because the exact role of these lymphocytes is unclear, we investigated the mechanisms of migration and adhesion of TILs to bone metastatic tumors, particularly to osteoblasts and bone marrow-derived stromal cell(BMSC)s. Histopathological examination showed that most TILs in secondary bone metastatic tumors (from primary tumors in the lung or breast) were found in the supporting tissue stroma between the bone and tumor mass. Cultured TILs (obtained from breast tumors) adhered spontaneously to osteoblasts and BMSCs (obtained from patients with osteoarthritis) without exogenous stimulation. Adhesion was further enhanced by chemokines macrophage inflammatory protein (MIP) 1alpha and MIP-1beta. TILs highly expressed activation antigens CD25 and CD69. A spontaneous activation of an integrin, lymphocyte function-associated antigen-1 (LFA-1), was also detected on TILs. TILs produced high concentrations of MIP 1alpha and MIP-1beta and spontaneous polymerization of cytoskeletal F-actin was observed in these cells. Adhesion of TILs to osteoblasts and BMSCs via LFA-1 and very late antigen-4 was associated with the production of osteoclastogen interleukin 6 by the latter cells. Our results indicate that integrins on TILs are activated in an autocrine manner by MIP-1alpha and MIP-1beta, and that treatment with the chemokines increases the binding of TILs on osteoblasts and stromal cells via a mechanism involving intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 as targets for the integrin. Our data also indicated that interactions between TILs and osteoblasts/stromal cells lead to the secretion by the latter of the osteoclastogenic cytokine interleukin 6. PMID- 9751627 TI - Targeting of interleukin 2 to human ovarian carcinoma by fusion with a single chain Fv of antifolate receptor antibody. AB - To provide a new tool for the immunotherapy of human ovarian carcinoma, we constructed a fusion protein between interleukin-2 (IL-2) and the single-chain Fv (scFv) of MOV19, a monoclonal antibody directed against alpha-folate receptor (alpha-FR), known to be overexpressed on human nonmucinous ovarian carcinoma. This was accomplished by fusing the coding sequences in a single open reading frame and expressing the IL-2/MOV19 scFv chimera under the control of the murine immunoglobulin K promoter in J558L plasmacytoma cells. The design allowed the construction of a small molecule combining the specificity of MOV19 with the immunostimulatory activity of IL-2. This might improve the tissue penetration and distribution of the fusion protein within the tumor, reduce its immunogenicity, and avoid the toxicity related to the systemic administration of IL-2. The IL 2/MOV19 fusion protein was stable on purification from the cell supernatant and was biologically active. Importantly, this construct was able to target IL-2 onto the surface of alpha-FR-overexpressing tumor cells and stimulated the proliferation of the IL-2-dependent CTLL-2 cell line as well as that of human resting peripheral blood lymphocytes. In a syngeneic mouse model, IL-2/MOV19 scFv specifically targeted a-FR gene-transduced metastatic tumor cells without accumulating in normal tissues, due to its fast clearance from the body. Prolonged release of IL-2/MOV19 scFv by in vivo transplanted J558-EF6.1 producer cells protected 60% of mice from the development of lung metastases caused by an i.v. injection of a-FR gene-transduced tumor cells. Moreover, treatment with IL 2/MOV19 scFv, but not with recombinant IL-2, significantly reduced the volume of s.c. tumors. The pharmacokinetics and biological characteristics of IL-2/NMOV19 scFv might allow us to combine the systemic administration of this molecule with the adoptive transfer of in vitro retargeted T lymphocytes for the treatment of ovarian cancer, thereby providing local delivery of IL-2 without toxicity. PMID- 9751628 TI - Somatic mutation of TSSC5, a novel imprinted gene from human chromosome 11p15.5. AB - We previously reported the isolation of a 2.5 Mb tumor-suppressing subchromosomal transferable fragment (STF) from 11p15.5 and the identification of nine known genes and four novel genes within this STF. We now report the isolation of a fifth novel cDNA, tumor-suppressing STF cDNA 5, designated TSSC5, located within the STF. TSSC5 encodes a predicted protein of 424 amino acids. Sequence analysis suggests that TSSC5 is a membrane protein with 10 transmembrane segments, and it is located between two imprinted genes, p57KIP2 and TSSC3. Northern blot hybridization revealed a 1.6-kb transcript in multiple adult tissues and in fetal liver and kidney, consistent with a potential role in embryonal tumors. We also found that TSSC5 is imprinted with preferential expression from the maternal chromosome. Reverse transcription-PCR analysis of TSSC5 revealed frequent occurrence of aberrant RNA splicing, which deleted exons 4, 5, and 6 in Wilms' tumors. Mutational analysis of TSSC5 by direct DNA sequencing of exons revealed a base substitution of G1120A in a Wilms' tumor, matched normal kidney, and the patient's mother, changed Arg at codon 309 to Gln. The G1120A substitution thus represents either a rare polymorphism or a tumor-predisposing mutation, because the mutant allele was of maternal origin and preferentially expressed in the patient's tissue. A second base substitution, C892T, was found in a lung cancer, changing Ser at codon 233 to Phe. This substitution was absent from the matched normal tissue and thus represented a somatic mutation. We also found loss of heterozygosity in the lung cancer, suggesting that TSSC5 may be a conventional tumor suppressor gene in the adult human lung and an imprinted tumor suppressor gene in the fetal kidney. PMID- 9751629 TI - Taxol resistance mediated by transfection of the liver-specific sister gene of P glycoprotein. AB - The sister gene of P-glycoprotein (Spgp) is a liver-specific ATP-binding cassette protein highly related to the P-glycoprotein (Pgp) family (S. Childs et al, Cancer Res., 55: 2029-2034, 1995). Spgp appears to be related to the Pgp family by an ancient duplication occurring before the division of fish and mammals. P Glycoproteins have diverse functions including broad specificity multidrug resistance in cell lines and tumors, detoxification of tissues such as the intestine and blood-brain barrier, and phosphatidylcholine transport in liver. Spgp is a Mr approximately 170,000 glycosylated plasma membrane protein localized to the canalicular surface of hepatocytes in the rat liver. The full-length cDNA of Spgp was isolated from rat, and its expression was characterized in situ and in transfected cells. The expression of Spgp correlates with the differentiation of hepatocytes and is seen only in late liver development. It is not observed in hepatoma cell lines. The physiological function of Spgp in liver is unknown, but it maps to 2q31 in humans, in the vicinity of liver transport disorders for bile acids and cholesterol. Spgp may therefore be involved in some aspect of bile acid or cholesterol metabolism. Spgp transfectants have a low level resistance to Taxol but not to other drugs that form part of the multidrug resistance phenotype. This resistance is reversible by the Pgp-reversing agents cyclosporin A, PSC833, and verapamil, suggesting a conservation in some functions of Pgps across large evolutionary distance. PMID- 9751630 TI - Telomerase activity in human development is regulated by human telomerase reverse transcriptase (hTERT) transcription and by alternate splicing of hTERT transcripts. AB - The correlation between telomerase activity, telomere lengths, and cellular replicative capacity has led to the theory that maintenance of telomere lengths by telomerase acts as a molecular clock to control replicative capacity and senescence. Regulation of this molecular clock may have applications in the treatment of cell aging and tumorigenesis, although little is presently known about the regulation of telomerase activity. To investigate possible mechanisms of regulation, we examined telomerase activity and the expression of the human telomerase RNA subunit and the human telomerase reverse transcriptase protein (hTERT) during human fetal heart, liver, and kidney development. The human telomerase RNA subunit is expressed in all three tissues at all gestational ages examined. hTERT expression correlates with telomerase activity in the liver, where both are expressed at all ages surveyed, and in the heart, where both are present until the 11th gestational week but not thereafter. However, although telomerase activity in the kidney is suppressed after the 15th gestational week, the hTERT transcript can be detected until at least the 21st week. Reverse transcription-PCR using primers within the reverse transcriptase domain of hTERT show the presence of multiple alternately spliced transcripts in these tissues, corresponding to full-length message as well as spliced messages with critical reverse transcriptase motifs deleted. Of note, telomerase activity in the kidney is only present at those gestational ages when full-length hTERT message is expressed (until approximately week 15), with spliced transcripts continuing to be expressed at later stages of development. The tissue-specific and gestational age dependent expression of hTERT mRNA seen in human development suggests the presence of at least two regulatory mechanisms controlling the activity of telomerase: transcriptional control of the hTERT gene and alternate splicing of hTERT transcripts. PMID- 9751631 TI - Frequency of prolonged remission duration after high-dose cytarabine intensification in acute myeloid leukemia varies by cytogenetic subtype. AB - Advances in the treatment of acute myeloid leukemia (AML) have occurred with the introduction of new therapies including high-dose cytarabine and the identification of powerful prognostic factors such as cytogenetics that predict for long-term outcome. To date, the prognostic impact of cytarabine dose escalation within various cytogenetic groups of AML has not been assessed. We describe 285 newly diagnosed patients with primary AML who had adequate karyotypes and were enrolled on a prospective Cancer and Leukemia Group B cytogenetic study. All patients were randomly assigned to postremission treatment with standard-, intermediate-, or high-dose cytarabine intensification. Patients were categorized to one of three cytogenetic groups: (a) core binding factor type [(CBF); ie., t(8;21) inv(16), t(16;16), and del(16)]; (b) normal; and (c) other abnormality karyotype. An evaluation of these patients after a median follow-up time of over 7 years was performed to determine the relationship of intensification to outcome by cytogenetic group. Patients included 57 patients with CBF AML, 140 patients with normal karyotype AML, and 88 patients with other cytogenetic abnormalities. The treatment outcome of CBF AML patients was superior, with an estimated 50% still in complete remission (CR) after 5 years as compared with 32 and 15% for patients with normal karyotype AML and other abnormality AML, respectively (P < 0.001). Univariate analysis showed the following nonkaryotype factors to predict a prolonged CR duration: (a) younger age (P < 0.008); (b) lower leukocyte count (P=0.01); (c) the presence of Auer rods (P=0.004); (d) a lower percentage of bone marrow blasts (P=0.001) at the time of diagnosis, (e) and a higher postremission cytarabine dose (P < 0.001). The impact of cytarabine dose on long-term remission was most marked (P < 0.001) in the CBF AML group (after 5 years, 78% of those with a dose of 3 g/m2 were still in CR, 57% of those with a dose of 400 mg/m2 were still in CR, and 16% of those with a dose of 100 mg/m2 were still in CR) followed by normal karyotype AML (P=0.01; after 5 years, 40% of those with a dose of 3 g/m2 were still in CR, 37% of those with a dose of 400 mg/m2 were still in CR, and 20% of those with a dose of 100 mg/m2 were still in CR). In contrast, cytarabine at all doses produced only a 21% or less chance of long-term continuous CR for patients with other cytogenetic abnormalities. A multivariate analysis of CR duration assessed the independent impact of each of these variables on cure. Significant factors entering this model in descending order of importance were cytogenetic group (CBF > normal > other abnormality; P=0.00001), cytarabine dose (3 g/m2 > 400 mg/m2 > 100 mg/m2; P=0.00001), logarithm of leukocyte count at the time of diagnosis (P=0.0005), and histological subtype of AML (P=0.005). This study demonstrates that the curative impact of cytarabine intensification varies significantly among cytogenetic groups and results in a substantial prolongation of CR among patients with CBF and normal karyotypes, but not in those with other karyotypic abnormalities. These findings support the use of pretreatment cytogenetics in risk stratification of postremission AML therapy. PMID- 9751632 TI - A point mutation within exon 5 of the WT1 gene of a sporadic unilateral Wilms' tumor alters gene function. AB - The Wilms' tumor suppressor gene, wt1, encodes a zinc-finger transcription factor, WT1, that represses transcription of a number of growth-promoting genes and inhibits cell growth. The transcripts of wt1 undergo two alternative splicing events, giving rise to four isoforms of mRNA in constant ratios. The first alternative splice introduces an extra exon 5, which encodes 17 amino acid residues inserted between the transcription regulatory domain and the DNA binding domain of WT1. Previously, we demonstrated that the 17-amino acid domain functioned as a transcription repressor when it was fused with the DNA binding domain of WT1. We have now identified a point mutation within exon 5 of wt1 in a sporadic unilateral Wilms' tumor patient. The mutation changes the last of the 17 amino acids from asparagine to serine. The protein isoform of WT1 carrying this mutation exhibited a 2-3-fold lower transcription-repressing activity than wild type WT1 in transient cotransfection assays. The mutation also decreased growth inhibiting activity of WT1 in two osteosarcoma cell lines, U2OS and Saos-2. By diminishing transcription-repressing and growth-inhibiting activities of WT1, this naturally occurring mutation within exon 5 of wt1 may disturb the normal function of the protein and lead to the uncontrolled cell growth characteristic of Wilms' tumor. PMID- 9751633 TI - Tumor growth modulation by sense and antisense vascular endothelial growth factor gene expression: effects on angiogenesis, vascular permeability, blood volume, blood flow, fluorodeoxyglucose uptake, and proliferation of human melanoma intracerebral xenografts. AB - Vascular endothelial growth factor (VEGF), also known as vascular permeability factor, has been investigated as a potent mediator of brain tumor angiogenesis and tumor growth. We evaluated the effect of VEGF expression on the pathophysiology of tumor growth in the brain. Human SK-MEL-2 melanoma cells, with minimal VEGF expression, were stably transfected with either sense or antisense mouse VEGF cDNA and used to produce intracerebral xenografts. Vascular permeability, blood volume, blood flow, and tumor fluorodeoxyglucose metabolism were assessed using tissue sampling and quantitative autoradiography. Tumor proliferation was assessed by measuring bromodeoxyuridine labeling indices. Tumor vascular density and morphological status of the blood-brain barrier were evaluated by immunohistochemistry. SK-MEL-2 cells transfected with sense VEGF (V+) expressed large amounts of mouse and human VEGF protein; V+ cells formed well-vascularized, rapidly growing tumors with minimal tumor necrosis. V+ tumors had substantial and significant increases in blood volume, blood flow, vascular permeability, and fluorodeoxyglucose metabolism compared to wild-type and/or V- (antisense VEGF) tumors. VEGF antisense transfected V- expressed no detectable VEGF protein and formed minimally vascularized tumors. V- tumors had a very low initial growth rate with central necrosis; blood volume, blood flow, vascular permeability, and glucose metabolism levels were low compared to wild-type and V+ tumors. A substantial inhibition of intracerebral tumor growth, as well as a decrease in tumor vascularity, blood flow, and vascular permeability may be achieved by down-regulation of endogenous VEGF expression in tumor tissue. VEGF targeted antiangiogenic gene therapy could be an effective component of a combined strategy to treat VEGF-producing brain tumors. PMID- 9751634 TI - Detection of high mobility group I HMGI(Y) protein in the diagnosis of thyroid tumors: HMGI(Y) expression represents a potential diagnostic indicator of carcinoma. AB - Hyperplastic or neoplastic proliferative lesions of thyroid follicular epithelium consist of a spectrum, ranging from nodular hyperplasia to undifferentiated (anaplastic) carcinoma, and usually present as palpable thyroid nodules. Thyroid nodules are a common occurrence in the general population, but only a small proportion of them are eventually diagnosed as carcinoma. The difficulty in objectively identifying those thyroid nodules that are malignant to avoid unnecessary surgery, combined with the range and effectiveness of the available therapeutic options in those patients who do, indeed, have thyroid carcinoma, has prompted the search for tumor markers and prognostic indicators. The high mobility group I (HMGI) proteins represent a class of nuclear proteins involved in the regulation of chromatin structure and function. HMGI(Y), one of the members of this class, is expressed at high levels during embryogenesis and in malignant tumors but at generally low levels in normal adult human tissues. Previous work on a limited number of thyroid samples suggested that the detection of the HMGI(Y) proteins may provide a clinically useful diagnostic tool. To verify this assumption, we analyzed HMGI(Y) expression by a combination of immunohistochemistry and reverse transcription-PCR in 358 thyroid tissue samples that were representative of the spectrum of thyroid tumor pathology. HMGI(Y) was detectable in 18 of 19 follicular carcinomas, 92 of 96 papillary carcinomas, and 11 of 11 undifferentiated (anaplastic) carcinomas but in only 1 of 20 hyperplastic nodules, 44 of 200 follicular adenomas, and 0 of 12 normal tissue samples. The correlation between HMGI(Y) expression and a diagnosis of carcinoma was highly significant (P < 0.0001). We also prospectively collected and analyzed for HMGI(Y) expression by immunohistochemistry and reverse transcription-PCR in 12 fine needle aspiration biopsies from 10 patients who subsequently underwent surgical removal of a solitary thyroid nodule. HMGI(Y) was detectable only in the four fine needle aspiration biopsies, corresponding to the thyroid nodules that were definitively diagnosed as carcinomas after surgery (two follicular carcinomas and two papillary carcinomas). The remaining eight samples (six follicular adenomas and two samples consisting of normal follicular cells) were negative. The findings of this study confirm the differential expression of HMGI(Y) in thyroid neoplasia and indicate the HMGI(Y) protein as a potential marker for thyroid carcinoma. PMID- 9751636 TI - Deciphering cryptic similarities in protein binding sites. AB - Rapid expansion in the number of plausible drug targets arising from genomics research has created new pressures for increased efficiency in discovery of high specificity candidate drug compounds. Improved understanding of conserved features among protein structures provides a promising route to achieving this goal. Indirect evidence implies that important similarities are now ripe for elucidation by emerging experimental approaches. PMID- 9751635 TI - Interleukin 4 inhibits growth and induces apoptosis in human breast cancer cells. AB - Interleukin-4 (IL-4) is a pleiotropic cytokine produced by mast cells and T lymphocytes that promotes proliferation and immunoglobulin class-switching in B cells. IL-4 receptors (IL-4Rs) are also expressed by nonhematopoietic cells as well as some tumor cells. Unlike its mitogenic effect on B cells, IL-4 inhibits the growth of some cancer cells in vitro. In this study, we show that IL-4R is expressed by breast and ovarian cancer cell lines. Furthermore, anchorage dependent and -independent growth of breast cancer cell lines MCF-7 and MDA-MB 231 is inhibited by IL-4 treatment, and this effect requires IL-4R. Interestingly, IL-4 only inhibited proliferating breast cancer cells and had no effect on basal, unstimulated growth. We therefore characterized the effect of IL 4 on breast cancer cell growth stimulated by either estradiol or insulin-like growth factor I (IGF-I). In both anchorage-dependent and -independent growth assays, IL-4 inhibited estradiol-stimulated growth. The antiestrogen effect of IL 4 was not due to IL-4 interference with the estrogen receptor, because IL-4 did not interfere with estrogen receptor-mediated reporter gene transactivation. In contrast, IL-4 had no effect on IGF-I-stimulated proliferation. Because IGF-I is known to inhibit programmed cell death, we examined apoptosis as a possible mechanism of IL-4 action. We established that IL-4 induced apoptosis in breast cancer cells by five independent criteria: (a) morphological indicators including pyknotic nuclei and cytoplasmic condensation; (b) DNA fragmentation; (c) the formation of DNA laddering; (d) the cleavage of poly(ADP-ribose) polymerase; and (e) the presence of cells with sub-G1 DNA content. IL-4 increased the percentage of apoptotic cells in MCF-7 and MDA-MB-231 cells 6.0- and 6.7-fold over that of the control, respectively. Finally, the addition of IGF-I reversed IL-4-induced apoptosis, suggesting that the mechanism of IL-4-induced growth inhibition in human breast cancer cells is the induction of programmed cell death. PMID- 9751637 TI - Engineering of modular polyketide synthases to produce novel polyketides. AB - Polyketides are important natural products produced by Actinomycetes and other organisms via the polymerization of coenzyme A-activated carboxylic acids. Modular polyketide synthases are large multifunctional enzymes that direct the biosynthetic process using a dedicated 'module' for each polymerization reaction, which specifies the unit to be polymerized, its oxidation state and stereochemistry. Over the past two years proof-of-principle has been demonstrated for technologies that modify or exchange modules to create hybrid enzymes that catalyze the biosynthesis of novel polyketides. PMID- 9751638 TI - Protein structure prediction. AB - Genome sequencing projects continue to provide a flood of new protein sequences, and prediction methods remain an important means of adding structural information. Recently, there have been advances in secondary structure prediction, which feed, in turn, into improved fold recognition algorithms. Finally, there have been technical improvements in comparative modelling, and studies of the expected accuracy of three-dimensional structural models built by this method. PMID- 9751639 TI - The development of new biotechnologies using metalloprotein design. AB - A variety of methodologies are under development to alter the behavior of existing metal centers or create entirely new sites within a protein framework in order to exploit the intrinsic chemical versatility of metals using the exquisite level of control that a protein matrix can exert to modulate their reactivity. Even at this relatively early stage, engineering of metal centers has led to the development of a number of emerging technologies with a wide variety of applications, including affinity purification of proteins, engineering of metal mediated protein stability, control of protein activity, imaging and therapy, biosensors, and new catalysts. PMID- 9751640 TI - Membrane protein structures: the known world expands. AB - The structure determinations of the cytochrome bc1 complex and the prokaryotic potassium channel demonstrate that a wider range of membrane proteins are now amenable to study by X-ray crystallography. Furthermore, the structures of porins and interfacial membrane proteins show that membrane structural biology is becoming a mature and productive field. PMID- 9751641 TI - Protein-mediated base flipping. AB - Since the discovery that the DNA methyltransferase M.HhaI utilises a base flipping mechanism to expose its target cytosine during catalysis, this phenomenon has been observed in other nucleic acid modifying enzymes. The crystallographic analyses of such enzyme-DNA complexes have revealed the molecular features of extrahelical base stabilisation, but have been less informative about the flipping process itself. PMID- 9751642 TI - Stereochemistry of the chloroperoxidase active site: crystallographic and molecular-modeling studies. AB - BACKGROUND: Chloroperoxidase (CPO) is the most versatile of the known heme enzymes. It catalyzes chlorination of activated C-H bonds, as well as peroxidase, catalase and cytochrome P450 reactions, including enantioselective epoxidation. CPO contains a proximal heme-thiolate ligand, like P450, and polar distal pocket, like peroxidase. The substrate-binding site is formed by an opening above the heme that enables organic substrates to approach the activated oxoferryl oxygen atom. CPO, unlike other peroxidases, utilizes a glutamate acid-base catalyst, rather than a histidine residue. RESULTS: The crystal structures of CPO complexed with exogenous ligands, carbon monoxide, nitric oxide, cyanide and thiocyanate, have been determined. The distal pocket discriminates ligands on the basis of size and pKa. The refined CPO-ligand structures indicate a rigid active-site architecture with an immobile glutamate acid-base catalyst. Molecular modeling and dynamics simulations of CPO with the substrate cis-beta methylstyrene and the corresponding epoxide products provide a structural and energetic basis for understanding the enantioselectivity of CPO-catalyzed epoxidation reactions. CONCLUSIONS: The various CPO-ligand structures provide the basis for a detailed stereochemical mechanism of the formation of the intermediate compound I, in which Glu183 acts as an acid-base catalyst. The observed rigidity in the active site also explains the relative instability of CPO compound I and the formation of the HOCI chlorinating species. Energetics of CPO-substrate/ product molecular modeling provides a theoretical basis for the P450-type enantioselective epoxidation activities of CPO. PMID- 9751643 TI - Substrate specificity of prostate-specific antigen (PSA). AB - BACKGROUND: The serine protease prostate-specific antigen (PSA) is a useful clinical marker for prostatic malignancy. PSA is a member of the kallikrein subgroup of the (chymo)trypsin serine protease family, but differs from the prototypical member of this subgroup, tissue kallikrein, in possessing a specificity more similar to that of chymotrypsin than trypsin. We report the use of two strategies, substrate phage display and iterative optimization of natural cleavage sites, to identify labile sequences for PSA cleavage. RESULTS: Iterative optimization and substrate phage display converged on the amino-acid sequence SS(Y/F)Y decreases S(G/S) as preferred subsite occupancy for PSA. These sequences were cleaved by PSA with catalytic efficiencies as high as 2200-3100 M-1 s-1, compared with values of 2-46 M-1 s-1 for peptides containing likely physiological target sequences of PSA from the protein semenogelin. Substrate residues that bind to secondary (non-S1) subsites have a critical role in defining labile substrates and can even cause otherwise disfavored amino acids to bind in the primary specificity (S1) pocket. CONCLUSION: The importance of secondary subsites in defining both the specificity and efficiency of cleavage suggests that substrate recognition by PSA is mediated by an extended binding site. Elucidation of preferred subsite occupancy allowed refinement of the structural model of PSA and should facilitate the development of more sensitive activity-based assays and the design of potent inhibitors. PMID- 9751644 TI - Homologs of the vancomycin resistance D-Ala-D-Ala dipeptidase VanX in Streptomyces toyocaensis, Escherichia coli and Synechocystis: attributes of catalytic efficiency, stereoselectivity and regulation with implications for function. AB - BACKGROUND: Vancomycin-resistant enterococci are pathogenic bacteria that have altered cell-wall peptidoglycan termini (D-alanyl-D-lactate [D-Ala-D-lactate] instead of D-alanyl-D-alanine [D-Ala-D-Ala]), which results in a 1000-fold decreased affinity for binding vancomycin. The metallodipeptidase VanX (EntVanX) is key enzyme in antibiotic resistance as it reduces the cellular pool of the D Ala-D-Ala dipeptide. RESULTS: A bacterial genome search revealed vanX homologs in Streptomyces toyocaensis (StoVanX), Escherichia coli (EcoVanX), and Synechocystis sp. strain PCC6803 (SynVanX). Here, the D,D-dipeptidase catalytic activity of all three VanX homologs is validated, and the catalytic efficiencies and diastereoselectivity ratios for dipeptide cleavage are reported. The ecovanX gene is shown to have an RpoS (sigma(s))-dependent promoter typical of genes turned on in stationary phase. Expression of ecovanX and an associated cluster of dipeptide permease genes permitted growth of E. coli using D-Ala-D-Ala as the sole carbon source. CONCLUSIONS: The key residues of the EntVanX active site are strongly conserved in the VanX homologs, suggesting their active-site topologies are similar. StoVanX is a highly efficient D-Ala-D-Ala dipeptidase; its gene is located in a vanHAX operon, consistent with a vancomycin-immunity function. StoVanX is a potential source for the VanX found in gram-positive enterococci. The catalytic efficiencies of D-Ala-D-Ala hydrolysis for EcoVanX and SynVanX are 25-fold lower than for EntVanX, suggesting they have a role in cell-wall turnover. Clustered with the ecovanX gene is a putative dipeptide permease system that imports D-Ala-D-Ala into the cell. The combined action of EcoVanX and the permease could permit the use of D-Ala-D-Ala as a bacterial energy source under starvation conditions. PMID- 9751645 TI - The deoxyxylulose phosphate pathway of terpenoid biosynthesis in plants and microorganisms. AB - Recent studies have uncovered the existence of an alternative, non-mevalonate pathway for the formation of isopentenyl pyrophosphate and dimethylallyl pyrophosphate, the two building blocks of terpene biosynthesis. PMID- 9751647 TI - DNA-enhanced peroxidase activity of a DNA-aptamer-hemin complex. AB - BACKGROUND: In vitro selection (SELEX) previously identified short single stranded DNAs that specifically bound N-methylmesoporphyrin IX (NMM), a stable transition-state analogue for porphyrin-metallation reactions. Interestingly, iron(III)-protoporphyrin (hemin) was a good competitive inhibitor for the DNA catalyzed metallation reaction, and appeared to bind strongly to the NMM-binding DNA aptamers. We investigated the peroxidase activity of the aptamer-hemin complexes to see if the DNA component of the complex, like the apoenzymes in protein peroxidases, could enhance the low intrinsic peroxidatic activity of hemin. RESULTS: Two porphyrin-binding DNA aptamers bound hemin with submicromolar affinity. The aptamer-hemin complexes had significantly higher peroxidase activity than hemin alone, under physiological conditions. The Vobs of the PS2.M hemin complex was 250 times greater than that of hemin alone, and significantly superior to a previously reported hemin-catalytic-antibody complex. Preliminary spectroscopic evidence suggests the coordination of the hemin iron in the complex changes, such that the complex more closely resembles horseradish peroxidase and other heme proteins rather than hemin. CONCLUSIONS: A new class of catalytic activity for nucleic acids is reported. The aptamer-hemin complexes described are novel DNA enzymes and their study will help elucidate the structural and functional requirements of peroxidase enzymes in general and the ways that a nucleic acid 'apoenzyme' might work to enhance the intrinsic peroxidatic ability of hemin. These aptamer-hemin complexes could be regarded as prototypes for redox catalyzing ribozymes in a primordial 'RNA world'. PMID- 9751646 TI - Combination of DMT-mononucleotide and Fmoc-trinucleotide phosphoramidites in oligonucleotide synthesis affords an automatable codon-level mutagenesis method. AB - BACKGROUND: Synthetic DNA has been used to introduce variability into protein coding regions. In protocols that produce a few mutations per gene, the sampling of amino-acid sequence space is limited by the bias imposed by the genetic code. It has long been apparent that the incorporation of trinucleotides in the synthetic regime would circumvent this problem and significantly enhance the usefulness of the technique. RESULTS: A new method is described for the creation of codon-level degenerate oligodeoxyribonucleotides that combines conventional dimethoxytrityl (DMT) mononucleoside phosphoramidite chemistry with 9 fluorenylmethoxycarbonyl (Fmoc) trinucleotide phosphoramidites (whose synthesis is reported in the paper). The substoichiometric use of these Fmoc-trinucleotides in an automatable, solid-phase synthesis procedure afforded DNA fragments comprising the wild-type sequence and a controllable distribution of mutants within two- and three-codon stretches of DNA, within the multiple cloning site of the conventional cloning vector pUC19. CONCLUSIONS: DMT and Fmoc are compatible protecting groups in conventional oligonucleotide synthesis methods, resulting in controllable levels of codon-based mutagenesis. PMID- 9751649 TI - Immunity to infection. PMID- 9751648 TI - Rhyme or reason: RNA-arginine interactions and the genetic code. AB - Theories about the origin of the genetic code require specific recognition between nucleic acids and amino acids at some stage of the code's evolution. A statistical analysis of arginine-binding RNA aptamers now offers the opportunity to test such interactions and provides the strongest support for an intrinsic affinity between any amino acid and its codons. PMID- 9751650 TI - Genetic effects on immunity. PMID- 9751651 TI - Mechanisms of central color vision. AB - In monkey cerebral cortex, color information is processed along the ventral visual pathway. This pathway starts in the primary visual cortex and ends in area TE of the inferior temporal cortex. Recent studies indicate that the transformation of cone signals occurs early in the pathway to form neurons selective to a narrow range of hues. In addition, it has become apparent that area TE plays a vital role in color discrimination. PMID- 9751652 TI - Cortical processing of complex sounds. AB - Work on the functional organization of auditory cortex in nonhuman primates has recently gained increasing attention. Neurophysiological studies using complex stimuli, combined with anatomical tract tracing, reveal a hierarchy of cortical processing comparable to other sensory systems. On the basis of these findings from animal studies, together with the advent of modern neuroimaging methods used in human cortex, the field of auditory neuroscience could soon arrive at a detailed understanding of the cortical representation of complex sounds, including speech. PMID- 9751653 TI - Complex microstructures of sensory cortical connections. AB - The neocortex has a distinctive laminar and modular organization. Although important questions remain regarding structure and function at this level of organization, recent studies are addressing a finer scale of synaptic and network microstructure. New findings concerning network properties are rapidly emerging from approaches in which dual or triple intracellular recordings in vitro are combined with analyses of cell and synaptic morphology, as well as from experiments designed to label multiple cell populations. PMID- 9751654 TI - Mechanisms of stereoscopic vision: the disparity energy model. AB - The past year has seen significant advances in our understanding of the role played by the primary visual cortex (V1) in stereoscopic vision. Recently, the mechanism by which complex cells in V1 respond to random-dot stereograms has been characterized; it appears that their response properties greatly reduce the complexity of one of the critical links for stereopsis, the correspondence problem. PMID- 9751655 TI - Hierarchical somatosensory processing. AB - Recent studies of the postcentral and additional somatosensory cortices support a hierarchical scheme for information processing. In the postcentral gyrus, the complexity of receptive field properties increases with caudal progression from area 1. It has been reported that the anterior bank of the intraparietal sulcus, the caudalmost part of the postcentral gyrus, is responsible for the systematic integration of bilateral body parts, as well as of somatic and visual information. PMID- 9751656 TI - Feedforward, horizontal, and feedback processing in the visual cortex. AB - The cortical visual system consists of many richly interconnected areas. Each area is characterized by more or less specific receptive field tuning properties. However, these tuning properties reflect only a subset of the interactions that occur within and between areas. Neuronal responses may be modulated by perceptual context or attention. These modulations reflect lateral interactions within areas and feedback from higher to lower areas. Recent work is beginning to unravel how horizontal and feedback connections each contribute to modulatory effects and what the role of these modulations is in vision. Whereas receptive field tuning properties reflect feedforward processing, modulations evoked by horizontal and feedback connections may reflect the integration of information that underlies perception. PMID- 9751657 TI - Gustatory mechanisms for the detection of fat. AB - Recent evidence suggests that free fatty acids may be one of the important stimuli used by taste receptor cells for the detection of fat. Consistent with this interpretation, the proteins necessary for the release and transport of lipophilic fatty acids are found in the oral cavity, and taste cells have recently been shown to contain fatty-acid-sensitive ion channels and transport molecules for the uptake of fatty acids. PMID- 9751658 TI - Phylogenetic development of the cochlea and its innervation. AB - Comparative studies of vertebrate hearing organs have enabled an integrated approach to difficult questions related to function. Recent evidence for the independent evolution of similar hearing-organ specializations, in particular hair-cell differentiation, has helped identify common problems of hearing receptors and put them in a new perspective. Evidence that cochlear amplification is an ancient phenomenon has widened the search for the motor mechanism involved. In this regard, different hypotheses are best examined by making optimal use of natural structural variations. Studies on the evolution of the efferent system have provided new routes to investigate its function. PMID- 9751659 TI - In vitro studies of cochlear excitation. AB - Although initially met with scepticism, the in vitro temporal bone preparation of the cochlea has proved to be a very important tool for investigating the function of the mammalian auditory system. As present techniques are able to maintain sufficient cellular viability, the in vitro preparation offers a valuable bridge between investigations using isolated outer hair cells and the intact system in vivo. PMID- 9751660 TI - Population coding in the retina. AB - Recent advances in multi-electrode recording have brought us closer to understanding how visual information is encoded by populations of retinal ganglion cells. By monitoring the visual responses of many ganglion cells at once, it is now possible to examine how ganglion cells act together to encode a visual scene. PMID- 9751661 TI - The specification of olfactory neurons. AB - Recent studies have shed light on the different relationships between odorant receptor expression and the specification of neural identity in the olfactory systems of vertebrates and invertebrates. In mice, neuronal identity and axon guidance are specified by the single expressed olfactory receptor, whereas in C. elegans, neuronal identity appears to be independent of receptor expression. PMID- 9751662 TI - Object vision and visual awareness. AB - Conscious experience involves perceiving, attending, remembering, and recognizing. Recent neuroscientific research has made significant contributions to our understanding of the mechanisms that mediate such capacities. Physiological and neuropsychological investigations have provided increasingly detailed descriptions of the location and functional properties of the brain structures involved in conscious perception, in attentive behavior and working memory, and in the recognition of objects. Such studies suggest that awareness of a visual stimulus probably reflects the interconnectivity and the type of cells involved in the representation of this stimulus, rather than the activity of specific circumscribed visual areas or processing streams. PMID- 9751663 TI - Complex motion perception and its deficits. AB - Within the hierarchy of motion perception, the dorsolateral middle superior temporal area (MSTd) is optimally suited for the analysis of the complex motion patterns that are directly useful for visually guided behaviour (e.g. computation of heading). Recent electrophysiological and psychophysical evidence suggests the existence of 'detectors' in MSTd that are specialised for complex motion patterns and advocates the necessity of combining retinal and extraretinal signals received by MSTd neurones for the accurate perception of heading. In some neurological patients, of which only a small number have been reported to date, lesions involving the human homologue of MST have devastating effects on their ability to navigate in their surroundings. It has been reported that these patients have impaired performance of psychophysical tasks of complex motion discrimination. PMID- 9751665 TI - Factors controlling hair-cell regeneration/repair in the inner ear. AB - Damaged hair cells in the avian basilar papilla are replaced by regenerative proliferation of supporting cells and transdifferentiation of supporting cells into hair cells. In the mammalian vestibular system, transdifferentiation and, possibly, the repair of damaged hair cells appear to play significant roles. Several growth factors have been found to be associated with the regeneration/repair process: insulin, insulin-like growth factor 1 (IGF-1), and fibroblast growth factors are important for avian inner ear regeneration/repair, whereas epidermal growth factor, transforming growth factor alpha, insulin, IGF 1, and IGF-2 are important for regeneration/repair in the mammalian labyrinth. Increasing evidence suggests that regeneration/repair of mammalian auditory hair cells is possible during the early neonatal period and may exist to a very limited degree at later times. PMID- 9751664 TI - Discrimination of pheromonal cues in fish: emerging parallels with insects. AB - Many fish species employ hormonal products as sex pheromones, and these cues are often mixtures that are released with a temporal pattern. This behavior is strikingly similar to that of insects, as moths use precise blends of odorants as sex pheromones and are skillful at tracking them in spite of changes in odor intensity associated with aerial dispersal. New studies in both groups of animals suggest many parallels in the functional anatomy of olfactory pathways and the organization of information-coding circuits. PMID- 9751666 TI - Sensory systems. AB - Research on the senses spans the enormous range from analysis of individual molecules involved in sensory transduction to the attempted elucidation of conscious sensation. Because the variety of conceptual and experimental approaches varies so broadly across the field, it is impossible to delineate a single direction for future research. Two trends are nonetheless apparent. At the reductionistic end of the spectrum, in the analysis of sensory transduction, studies on all the senses will increasingly be driven by the techniques of molecular biology. The advent of techniques for producing cDNA libraries from small ensembles of receptor cells, or even from individual cells, will permit the recognition of new constituents of receptor cells and of factors involved in their specification, differentiation, and maintenance. In the integrative realm of sensory neurobiology, future studies will increasingly rely on optical techniques for the study of activity patterns on the surfaces of sensory areas of the cerebral cortex and on noninvasive functional imaging for the investigation of neural responses in human subjects. These techniques will continue to strengthen our understanding of the relation between neuronal activity and conscious sensory experience. PMID- 9751667 TI - Mycobacterial lipids: a historical perspective. AB - Mycobacterial lipids have been studied for more than 70 years, due to the fascinating diversity of their structures and biological activities. A historical perspective, and the present status on the structure and activity of major lipids of the outer envelope of mycobacterial cells are presented : mycolic acids, which are main constituents of the cell wall, and glycolipids known for toxic or immunological properties (cord factor, SL, DAT, PGL, GPL, LOS, LAM). As far as possible, it was tried to distinguish between experimentally established knowledge and currently accepted speculations. PMID- 9751668 TI - The role of carbohydrates in mammalian sperm-egg interactions: how important are carbohydrate epitopes? AB - Evidence implicating the involvement of carbohydrates in fertilization has been reported for decades in species which span the phylogenetic scale. The exact nature and role of these ligands in fertilization, however, has eluded investigators. Here, such investigations are reviewed as they relate to mammalian fertilization, with the principle focus on reviewing the role of carbohydrates involved in the primary binding event between sperm cell and egg. PMID- 9751669 TI - Dilemmas of NIH funding for cardiovascular research. PMID- 9751670 TI - Milestones for Dr DeBakey. PMID- 9751671 TI - Vasodilator reserve: a functional assessment of coronary health. PMID- 9751673 TI - Delivery of colloidal particles and red blood cells to tissue through microvessel ruptures created by targeted microbubble destruction with ultrasound. AB - BACKGROUND: We have previously shown that the application of ultrasound to thin shelled microbubbles flowing through small microvessels (<7 microm in diameter) produces vessel wall ruptures in vivo. Because many intravascular drug- and gene delivery vehicles are limited by the endothelial barrier, we hypothesized that this phenomenon could be used to deliver drug-bearing vehicles to tissue. METHODS AND RESULTS: An exteriorized rat spinotrapezius muscle preparation was used. Intravascular fluorescent red blood cells and polymer microspheres (PM) (205 and 503 nm in diameter) were delivered to the interstitium of rat skeletal muscle through microvessel ruptures created by insonifying microbubbles in vivo. On intravital microscopy, mean dispersion areas per rupture for red blood cells, 503 nm PM, and 205-nm PM were 14.5x10(3) microm2, 24. 2x10(3) microm2, and 27.2x10(3) microm2, respectively. PM dispersion areas were significantly larger than the mean dispersion area for red blood cells (P<0.05). CONCLUSIONS: Microvessel ruptures caused by insonification of microbubbles in vivo may provide a minimally invasive means for delivering colloidal particles and engineered red blood cells across the endothelial lining of a targeted tissue region. PMID- 9751672 TI - Endothelium-dependent relaxation of collateral microvessels after intramuscular gene transfer of vascular endothelial growth factor in a rat model of hindlimb ischemia. AB - BACKGROUND: Recent investigations have demonstrated the ability of vascular endothelial growth factor (VEGF) to augment the development of collateral arteries in vivo. In vitro studies have suggested that the use of VEGF also improves the endothelium-dependent relaxation of collaterals at the microvascular level. The purpose of this study was to determine in vivo the extent to which vasomotor responses of collateral microvessels are altered after VEGF treatment. METHODS AND RESULTS: Ischemia was induced in the hindlimb of 35 rats by excision of the femoral artery. Immediately thereafter, 400 microg of a plasmid encoding VEGF or ss-galactosidase (control) was transfected into limb muscles. Four weeks later, synchrotron radiation microangiography, with a spatial resolution of 30 microm, was performed to document the reactivity of collateral microvessels. Administration of the endothelium-dependent vasodilator acetylcholine failed to induce dilation of collateral microvessels in control animals. By contrast, profound dilation of collaterals was observed after acetylcholine in VEGF-treated animals. This response was evident in vessels with a linear appearance but not in those with an undulating appearance. The resulting blood flow in the ischemic limb after administration of acetylcholine in the control animals was only 64.6+/ 17.0% of that of the contralateral normal limb, whereas blood flow was augmented to 106.1+/-8.4% in VEGF-treated animals (P<0.05). CONCLUSIONS: These results demonstrate in vivo that the use of VEGF restores impaired vasomotor responses in some types of collateral microvessels, which may help to provide a basis for understanding the microcirculation after therapeutic angiogenesis with VEGF. PMID- 9751674 TI - Pharmacodynamic efficacy, clinical safety, and outcomes after prolonged platelet Glycoprotein IIb/IIIa receptor blockade with oral xemilofiban: results of a multicenter, placebo-controlled, randomized trial. AB - BACKGROUND: Parenteral administration of platelet glycoprotein IIb/IIIa (GP IIb/IIIa) receptor blockers can reduce ischemic complications of coronary angioplasty. Orally active GP IIb/IIIa blockers may allow more sustained receptor antagonism with the potential for long-term secondary prevention. The pharmacodynamic efficacy, clinical safety, and outcomes after prolonged receptor blockade with an orally active GP IIb/IIIa antagonist are not known. The Oral Glycoprotein IIb/IIIa Receptor Blockade to Inhibit Thrombosis (ORBIT) Trial is a multicenter, placebo-controlled, randomized trial of xemilofiban, an oral platelet GP IIb/IIIa blocking agent, administered to patients after percutaneous coronary intervention. METHODS AND RESULTS: After successful elective percutaneous coronary intervention, 549 patients were randomized to receive either placebo or xemilofiban in a dose of 15 or 20 mg. Stented patients randomized to placebo also received ticlopidine 250 mg orally BID for 4 weeks. Patients who received abciximab during the coronary intervention and who were randomized to receive xemilofiban were administered a reduced dosage (10 mg TID for 2 weeks) followed by the randomized maintenance dose of 15 or 20 mg BID for 2 more weeks. All patients received 325 mg aspirin PO QD. Ex vivo platelet aggregation in response to 20 micromol/L ADP and 4 microg/mL collagen was measured over time after the initial dose of study drug and at days 14 and 28 of long-term therapy in 230 patients. All patients were followed clinically for 90 days. Xemilofiban inhibited platelet aggregation to both ADP and collagen with peak levels of inhibition that were similar at 14 and 28 days of long-term oral therapy. Plasma levels of xemilofiban correlated with the degree of platelet inhibition. Peak platelet inhibition on day 1 correlated with the subsequent occurrence of insignificant or mild bleeding events. Although this study was not powered to evaluate differences in clinical outcomes, a trend (P=0.04) was observed for reduction of cardiovascular events at 3 months in patients not treated with abciximab who received the highest dose (20 mg) of xemilofiban studied. CONCLUSIONS: Xemilofiban inhibited platelet aggregation and was well tolerated during 28 days of long-term oral therapy. The observed trend in reduction of cardiovascular events in follow-up awaits confirmation in the larger scale phase III study (EXCITE trial) currently in progress. PMID- 9751676 TI - Relation of Helicobacter pylori infection to 13-year mortality and incident ischemic heart disease in the caerphilly prospective heart disease study. AB - BACKGROUND: Associations have been suggested between Helicobacter pylori seropositivity, cardiovascular risk factors, and ischemic heart disease (IHD). The effect of this common infection on mortality is uncertain. METHODS AND RESULTS: Plasma specimens collected during 1979 to 1983 from 1796 men in Caerphilly, South Wales, were analyzed for IgG antibodies to H pylori. Cause of death and occurrence of incident IHD events were ascertained over an average of 13.7 years from death certificates, hospital records, and ECG changes at 5-yearly follow-up examinations. Seventy percent of men were seropositive. The prevalence of IHD at entry was similar in men with and without H pylori antibodies (odds ratio [OR], 1.10; 95% CI, 0.87 to 1.40). Seropositivity was significantly (P<0.05) associated with poorer socioeconomic status currently and in childhood, shorter stature, and poorer ventilatory function at entry but not with age, smoking, body mass index, blood pressure, total cholesterol, HDL cholesterol, LDL cholesterol, fibrinogen, plasma viscosity, or heat shock protein antibodies. Thirteen-year incidence of IHD was not significantly associated with H pylori (OR, 1.05; 95% CI, 0.80 to 1.39), but there was a stronger relationship with all cause mortality (OR, 1.46; 95% CI, 1.12 to 1.92) and fatal IHD (OR, 1.54; 95% CI, 1.03 to 2.30). After adjustment for cardiovascular risk factors and both adult and childhood socioeconomic status, ORs were slightly reduced and lost statistical significance (OR=1.32 [95% CI, 0.99 to 1.78] for all-cause mortality and OR=1.52 [95% CI, 0.99 to 2.34] for fatal IHD). CONCLUSIONS: H pylori infection is unlikely to be as strong a risk factor for IHD as some previous studies have suggested, but its relationship to mortality, including fatal IHD, deserves further investigation. The mechanism underlying these associations is unlikely to involve hypertension, circulating lipid profile, fibrinogen, or cross reacting antibodies to bacterial heat shock proteins. PMID- 9751675 TI - Better outcome for women compared with men undergoing coronary revascularization: a report from the bypass angioplasty revascularization investigation (BARI) AB - BACKGROUND: Numerous studies have shown that women undergoing coronary revascularization procedures do so at a higher risk for an adverse outcome compared with men. However, the impact of advances in technology and improvements in techniques on in-hospital and long-term outcome after revascularization in women is unclear. METHODS AND RESULTS: We evaluated 1829 patients with symptomatic multivessel coronary disease randomized to CABG or PTCA in the Bypass Angioplasty Revascularization Investigation (BARI), of whom 27% were women. As expected, women were older (64.0 versus 60.5 years), with more congestive heart failure (14% versus 7%), hypertension (68% versus 42%), treated diabetes mellitus (31% versus 15%), and unstable angina (67% versus 61%) than men but had similar preservation of left ventricular function and extent of multivessel disease. Women assigned to surgery received the same number of total grafts but fewer internal mammary artery grafts (72% versus 85%, P<0. 01), and those assigned to angioplasty had more intended lesions (76% versus 71%, P<0.01) successfully dilated than men. At an average of 5.4 years' follow-up, crude mortality rates were similar in women (12.8%) and men (12.0%). The Cox regression model adjusting for baseline differences revealed that women had a significantly lower risk of death (relative risk, 0.60; 95% CI, 0.43 to 0.84; P=0. 003) but not a significantly lower risk of death plus myocardial infarction (relative risk, 0.84; 95% CI, 0.66 to 1.07; P=0.16) than men. CONCLUSIONS: Although the unadjusted mortality rate suggests that women and men undergoing CABG and PTCA have a similar 5-year mortality, women have higher risk profiles; consequently, contrary to previous reports, female sex is an independent predictor of improved 5-year survival after we control for multiple risk factors. PMID- 9751677 TI - Effects of short-term treatment of hyperlipidemia on coronary vasodilator function and myocardial perfusion in regions having substantial impairment of baseline dilator reverse. AB - BACKGROUND: We tested the hypothesis that correction of hyperlipidemia improves coronary vasodilator response and maximal perfusion in myocardial regions having substantial impairment of pretreatment vasodilator capacity. METHODS AND RESULTS: Measurements of myocardial blood flow were made with PET [13N]ammonia in 12 patients with ischemic heart disease (11 men; age, 65+/-8 years [mean+/-SD]) at rest and during adenosine at 70 and then 140 microg . kg-1 . min-1 for 5 minutes each before and approximately 4 months after simvastatin treatment (40 mg daily). Simvastatin reduced LDL (171+/-13 before versus 99+/-18 mg/dL after simvastatin, P<0.001) and increased HDL (39+/-8 versus 45+/-9 mg/dL, P<0.05). Myocardial segments were classified on the basis of pretreatment blood flow response to 140 microg . kg-1 . min-1 adenosine as normal (flow >/=2 mL . min-1 . g-1) or abnormal (flow <2 mL . min-1 . g-1). In normal segments, baseline myocardial blood flow (0.95+/-0.32) increased (P<0.001) at both low- (1.62+/-0.81) and high- (2.63+/-0.41) dose adenosine and was unchanged both at rest and with adenosine after simvastatin. In abnormal segments, myocardial blood flow at rest (0. 73+/ 0.19) increased at low- (1.06+/-0.59, P<0.02) and high- (1. 29+/-0.33, P<0.01) dose adenosine. After simvastatin, myocardial blood flow increased more compared with pretreatment at both low- (1. 37+/-0.66, P<0.05 versus pretreatment) and high- (1.89+/-0.79, P<0. 01 versus pretreatment) dose adenosine. CONCLUSIONS: Short-term lipid-lowering therapy increases stenotic segment maximal myocardial blood flow by approximately 45%. The mechanism involves enhanced, flow-mediated dilation of stenotic epicardial conduit vessels and may account at least in part for the efficacy of lipid lowering in secondary prevention trials and in reducing ischemic episodes in ambulatory patients. PMID- 9751678 TI - Effect of high- and low-molecular-weight heparins on thrombin-thrombomodulin interaction and protein C activation. AB - BACKGROUND: Thrombin-thrombomodulin (TM) interaction, which is critical for accelerating the protein C anticoagulant pathway, involves the heparin-like domain of TM. This study was aimed at investigating the possible effect of heparin on thrombin-TM binding and protein C activation. METHODS AND RESULTS: The affinity of thrombin-TM interaction was studied by a functional method, based on the ability of thrombin-TM adduct to activate protein C, and by evaluation of the binding of thrombin to immobilized TM. Both experimental approaches showed that the affinity of thrombin-TM interaction was decreased by micromolar heparin concentrations. Heparin had no significant effect when a recombinant TM form, lacking the chondroitin sulfate moiety, was used. Furthermore, it was also shown that the inhibitory effect of heparin was directly proportional to the heparin molecular mass (molecular weight range, 3 to 16 kDa), which suggests that the effect was mediated by formation of electrostatic bonds between heparin and thrombin. CONCLUSIONS: These results indicate that heparin at therapeutic concentrations reduces the affinity of thrombin for TM and the rate of protein C activation. The magnitude of this effect is proportionally linked to the molecular mass of heparin. PMID- 9751679 TI - 1,25-Dihydroxyvitamin D3 increases in vitro vascular calcification by modulating secretion of endogenous parathyroid hormone-related peptide. AB - BACKGROUND: A significant association between vascular calcification and osteoporosis has been noted, suggesting that calcium homeostasis is important in vascular calcification as well as in osteoporosis. Moreover, results of our previous studies suggest that calcium-regulating hormones such as parathyroid hormone-related peptide (PTHrP) may modulate vascular calcification. Therefore, we hypothesized that 1alpha,25-dihydroxyvitamin D3 [1,25(OH)2D3] may have a direct impact on the calcium-regulating system of vascular smooth muscle cells, resulting in deposition of calcium in vascular wall. METHODS AND RESULTS: We investigated the effect of 1,25(OH)2D3 on in vitro calcification by bovine vascular smooth muscle cells (BVSMCs). 1,25(OH)2D3 dose dependently increased BVSMC calcification and alkaline phosphatase activity. 1,25(OH)2D3 also decreased secretion of PTHrP by BVSMCs in a dose-dependent manner and depressed its gene expression. Furthermore, exogenous PTHrP (fragment 1-34) antagonized the stimulatory effect of 1,25(OH)2D3 on BVSMCs. Finally, 1,25(OH)2D3 dose dependently increased the expression of the osteopontin gene, one of the bone matrix proteins in BVSMCs, contributing to its stimulatory action on BVSMC calcification. CONCLUSIONS: These data suggest that 1,25(OH)2D3 exerts a stimulatory effect on vascular calcification through direct inhibition of the expression of PTHrP in BVSMCs as an endogenous inhibitor of vascular calcification. Moreover, the stimulatory effects of 1,25(OH)2D3 on alkaline phosphatase activity and osteopontin expression may contribute to its promoting action in vascular calcification. PMID- 9751680 TI - New insights and observations in three-dimensional echocardiographic visualization of ventricular septal defects: experimental and clinical studies. AB - BACKGROUND: The positions, sizes, and shapes of ventricular septal defects (VSDs) can be difficult to assess by 2-dimensional echocardiography (2DE). Volume rendered 3-dimensional echocardiography (3DE) can provide unique views of VSDs from the left ventricular (LV) side, allowing complete assessment of their circumference and spatial orientations to other anatomic structures. METHODS AND RESULTS: Seventeen experimentally created defects of various locations, sizes, and shapes were imaged and reconstructed in 9 explanted porcine hearts. From an en face projection, major and minor axis diameters of the defects were measured, and these data were compared with direct anatomic measurements. Optimal reconstructions of the VSDs were obtained in all heart specimens, accurately depicting their positions and shapes. The correlations between 3DE and anatomy for the VSD major and minor axis diameters were y=1.0x+0.3 (r=0.88, P<0.001) and y=1.0x-1.4 (r =0.89, P<0.001), respectively. Good agreement between the 2 methods was demonstrated for all measurements. Our experience from the in vitro model was then applied to patient studies. Optimal LV en face reconstructions were obtained in 45 of 51 patients, permitting detailed assessment of the positions, sizes, and shapes of the VSDs. In the 25 patients with comparative surgical measurements, the correlations between 3DE and surgery for the VSD major and minor axis diameters were y =0. 81x+2.1 (r=0.92, P<0.001) and y=0.73x+2.0 (r=0.91, P<0.001), respectively. Good agreement was demonstrated between measurements made by 3DE and those obtained at surgery. CONCLUSIONS: 3DE provides excellent visualization of various types of VSDs. From an LV en face projection, the positions, sizes, and shapes of VSDs can be accurately determined. Such precise imaging will be beneficial for surgical and catheter-based closure of difficult perimembranous and singular or multiple muscular VSDs. PMID- 9751681 TI - Atrioventricular conduction during long-term follow-up of patients with sick sinus syndrome. AB - BACKGROUND: It has been claimed that patients with sick sinus syndrome have an increased risk of developing AV block, but this has never been assessed prospectively. The aim of the present study was to evaluate in a prospective trial AV conduction during the long-term follow-up of patients with sick sinus syndrome. METHODS: Two hundred twenty-five consecutive patients with sick sinus syndrome and intact AV conduction were randomized to undergo single-chamber atrial pacing (110 patients) or single-chamber ventricular pacing (115 patients). Follow-up after 3 months and then yearly included measurement of the PQ interval and, in patients with atrial pacemakers, determination of the atrial stimulus-Q intervals at pacing rates of 100 and 120 bpm. The occurrence of AV block in the atrial group was recorded. During follow-up (mean, 5.5+/-2.4 years), there was no change in PQ interval in either group and no change in atrial stimulus-Q intervals or Wenckebach block point in the atrial group. Four of 110 patients in the atrial group developed grade 2 to 3 AV block that required upgrading of the pacemaker (0.6% per year). Two of these 4 patients had right bundle-branch block at pacemaker implantation. CONCLUSIONS: AV conduction, estimated as PQ interval and atrial stimulus-Q interval at atrial pacing rates of 100 and 120 bpm and the Wenckebach block point, remains stable during long-term follow-up. Thus, treatment with single-chamber atrial pacing is safe and can be recommended to patients with sick sinus syndrome without bundle-branch block. PMID- 9751682 TI - Synergism between platelets and neutrophils in provoking cardiac dysfunction after ischemia and reperfusion: role of selectins. AB - BACKGROUND: Neutrophils (PMNs) are known to contribute to both cardiac dysfunction and myocardial necrosis after reperfusion of an ischemic heart. Moreover, platelets are also important blood cells that can aggravate myocardial ischemic injury. This study was designed to test the effects of PMNs and platelets separately and together in provoking cardiac dysfunction in isolated perfused rat hearts after ischemia and reperfusion. METHODS AND RESULTS: Control rat hearts not subjected to ischemia were perfused without blood cells for 80 minutes. Additional control rat hearts were perfused with 75x106 PMNs, with 100x106 platelets, or with 75x106 PMNs+100x106 platelets over a 5-minute perfusion followed by a 75-minute observation period. No significant reduction in coronary flow, left ventricular developed pressure (LVDP), or the first derivative of LVDP (dP/dtmax) was observed at the end of the observation period in any nonischemic group. Similarly, global ischemia (I) for 20 minutes followed by 45 minutes of reperfusion (R) produced no sustained effects on the final recovery of any of these parameters in any group of hearts perfused in the absence of blood cells. However, I/R hearts perfused with either PMNs or platelets alone exhibited decreases in these variables of 10% to 12% (P<0.05 from control). Furthermore, I/R hearts perfused with both PMNs and platelets exhibited decreases of 50% to 60% in all measurements of cardiac function (P<0.001). These dual-cell-perfused I/R hearts also exhibited marked increases in cardiac myeloperoxidase (MPO) activity, indicating a significant PMN infiltration, and enhanced P-selectin expression on the coronary microvascular endothelium. All cardiodynamic effects as well as MPO accumulation and PMN infiltration were markedly attenuated by a sialyl LewisX-oligosaccharide or a recombinant soluble P selectin ligand, which inhibits selectin-mediated cell adhesion. CONCLUSIONS: These results provide evidence that platelets and neutrophils act synergistically in provoking postreperfusion cardiac dysfunction and that this may be largely due to cell-to-cell interactions mediated by P-selectin. These findings may help explain the reperfusion injury phenomenon. PMID- 9751683 TI - Norepinephrine stimulates apoptosis in adult rat ventricular myocytes by activation of the beta-adrenergic pathway. AB - BACKGROUND: Myocardial sympathetic activity is increased in heart failure. We tested the hypothesis that norepinephrine (NE) stimulates apoptosis in adult rat ventricular myocytes in vitro. METHODS AND RESULTS: Myocytes were exposed to NE alone (10 micromol/L), NE+propranolol (2 micromol/L), NE+prazosin (0.1 micromol/L), or isoproterenol (ISO, 10 micromol/L) for 24 hours. NE and ISO decreased the number of viable myocytes by approximately 35%. This effect was completely blocked by the beta-adrenergic antagonist propranolol but was not affected by the alpha1-adrenergic antagonist prazosin. NE increased DNA laddering on agarose gel electrophoresis and increased the percentage of cells that were stained by terminal deoxynucleotidyl transferase-mediated nick end-labeling from 5.8+/-1. 0% to 21.0+/-2.3% (P<0.01; n=4). NE likewise increased the percentage of apoptotic cells with hypodiploid DNA content as assessed by flow cytometry from 7.8+/-0.7% to 16.7+/-2.2% (P<0.01; n=6), and this effect was abolished by propranolol but not prazosin. ISO and forskolin (10 micromol/L) mimicked the effect of NE, increasing the percentage of apoptotic cells to 14.7+/-1.9% and 14. 4+/-2.2%, respectively. NE-stimulated apoptosis was abolished by the protein kinase A inhibitor H-89 (20 micromol/L) or the voltage-dependent calcium channel blockers diltiazem and nifedipine. CONCLUSIONS: NE, acting via the ss-adrenergic pathway, stimulates apoptosis in adult rat cardiac myocytes in vitro. This effect is mediated by protein kinase A and requires calcium entry via voltage-dependent calcium channels. NE-stimulated apoptosis of cardiac myocytes may contribute to the progression of myocardial failure. PMID- 9751684 TI - Graft permeabilization facilitates gene therapy of transplant arteriosclerosis in a rabbit model. AB - BACKGROUND: Smooth muscle cell (SMC) replication plays a central role in the pathogenesis of transplant arteriosclerosis. One strategy to eliminate dividing cells is to express a herpesvirus thymidine kinase (tk) gene that phosphorylates the nucleoside analogue ganciclovir into a toxic form leading to cell killing. However, medial SMCs are resistant to gene transfer unless the artery undergoes deendothelialization. We hypothesized that manipulations that increase the "porosity" of the artery can make SMCs prone to gene transfer without denudation. METHODS AND RESULTS: In organ culture of rabbit aorta, longitudinal stretch and supraphysiological pressure applied for 3 hours during incubation with adenoviral vector facilitated gene transfer into medial SMCs without denudation. Of the SMCs, 10.2+/-3.8% expressed a reporter gene of human placental alkaline phosphatase (hpAP), whereas SMCs in control arteries did not express hpAP. To evaluate the feasibility of transgene expression in arterial grafts, we performed such permeabilization-assisted reporter gene transfer into aortas of donor Dutch Belted rabbits and transplanted them into carotid arteries of recipient New Zealand White rabbits. Unstretched transfected grafts were used as a control. SMCs expressed hpAP (7. 3+/-2.4% of cells in 2 days and 4.2+/-1.9% in 2 weeks) in stretched grafts only. In the next series of experiments, we transfected stretched grafts with ADV-tk and combined transplantation with systemic administration of ganciclovir. Stretched ADV-hpAP grafts were used as a control. In 2 weeks, the formation of intimal thickening in tk-expressing grafts was significantly reduced (P<0. 01) because of a decrease in proliferating SMCs. CONCLUSIONS: Manipulations within target tissues can enhance the efficiency of gene transfer into SMCs. Although mechanical permeabilization is clinically problematic, in principle, targeting SMC replication may provide a genetic approach to the treatment of transplant arteriosclerosis. PMID- 9751686 TI - Angiographic and ultrasonic evidence of plaque rupture causing myocardial infarction. PMID- 9751685 TI - Voltage-dependent calcium channel promoter restores baroreflex sensitivity in conscious dogs with heart failure. AB - BACKGROUND: The aim of this study was to determine the mechanism by which the calcium channel promoter BAY y 5959 affects the control of heart rate and baroreflex sensitivity in conscious dogs with pacing-induced heart failure (HF). METHODS AND RESULTS: We compared responses to BAY y 5959, which increases inotropy and decreases chronotropy, with those to norepinephrine (NE), which coincidentally exerts the same directional effects on inotropy and chronotropy, albeit through different mechanisms, in the presence and absence of ganglionic blockade both in control and in HF. Both BAY y 5959 and NE elicit direct effects on the heart and indirect effects through activation of reflexes, primarily the sinoaortic baroreceptor reflex. BAY y 5959 still reduced heart rate in dogs with arterial baroreceptor denervation, but not after ganglionic blockade. HF induced classic catecholamine desensitization to the inotropic effects of NE and blunted reflex bradycardia. In contrast, inotropic responses to BAY y 5959 were preserved in HF. Surprisingly, the autonomically mediated bradycardia induced by BAY y 5959 was also preserved in HF. Baroreflex sensitivity was assessed in control and in HF by pulse interval-systolic arterial blood pressure (PI/SAP) slopes constructed in response to pharmacological alterations in arterial pressure. HF depressed the PI/SAP slope from 11.5+/-1.3 to 4.8+/-0.9 ms/mm Hg, but during BAY y 5959 infusion in HF, the PI/SAP slope was restored to 24.1+/-5.2 ms/mm Hg. To assess central versus peripheral actions of BAY y 5959, the agent was infused with intra carotid artery perfusion at a low dose, which acted centrally but did not have an effect peripherally. Under these conditions, it still decreased heart rate and restored baroreflex sensitivity (PI/SAP slope, 12.7+/-2.8 ms/mm Hg). CONCLUSIONS: Thus, the calcium promoter restores arterial baroreflex sensitivity in HF. Based on intra-carotid artery experiments, this occurs through a central nervous system and vagal mechanism. PMID- 9751687 TI - A broken heart. PMID- 9751688 TI - Antioxidants and nitrate tolerance. PMID- 9751689 TI - Metabolism of fatty acids and glucose. PMID- 9751690 TI - Viability assessment before CABG. PMID- 9751691 TI - Effect of age adjustment in predicting outcome. PMID- 9751692 TI - The Chinese human genome diversity project. PMID- 9751693 TI - In an immunological twilight zone. PMID- 9751694 TI - Native secondary structure formation in RNA may be a slave to tertiary folding. PMID- 9751695 TI - What else can the immune system recognize? PMID- 9751696 TI - Reconsidering targeted toxins to eliminate HIV infection: you gotta have HAART. AB - The success of highly active anti-retroviral therapy (HAART) has inspired new concepts for eliminating HIV from infected individuals. A major obstacle is the persistence of long-lived reservoirs of latently infected cells that might become activated at some time after cessation of therapy. We propose that, in the context of treatment strategies to deliberately activate and eliminate these reservoirs, hybrid toxins targeted to kill HIV-infected cells be reconsidered in combination with HAART. Such combinations might also prove valuable in protocols aimed at preventing mother-to-child transmission and establishment of infection immediately after exposure to HIV. We suggest experimental approaches in vitro and in animal models to test various issues related to safety and efficacy of this concept. PMID- 9751697 TI - Drug concentration heterogeneity facilitates the evolution of drug resistance. AB - Pathogenic microorganisms use Darwinian processes to circumvent attempts at their control through chemotherapy. In the case of HIV-1 infection, in which drug resistance is a continuing problem, we show that in one-compartment systems, there is a relatively narrow window of drug concentrations that allows evolution of resistant variants. When the system is enlarged to two spatially distinct compartments held at different drug concentrations with transport of virus between them, the range of average drug concentrations that allow evolution of resistance is significantly increased. For high average drug concentrations, resistance is very unlikely to arise without spatial heterogeneity. We argue that a quantitative understanding of the role played by heterogeneity in drug levels and pathogen transport is crucial for attempts to control re-emergent infectious disease. PMID- 9751698 TI - Cytotoxic agents directed to peptide hormone receptors: defining the requirements for a successful drug. AB - In principle, cell surface receptors that are overexpressed in tumor tissue could serve as targets for anticancer drugs attached to receptor ligands. The purpose of this paper is to identify the necessary elements for a successful receptor targeted drug. We used the gastrin/cholecystokinin type B receptor as a model delivery system, and we report on the synthesis, trafficking, and in vitro and in vivo evaluation of heptagastrin, the C-terminal heptapeptide of gastrin, linked via an appropriate linker to a potently cytotoxic ellipticine derivative, 1-[3-[N (3-aminopropyl)-N-methylamino]propyl]amino-9-methoxy-5, 11-dimethyl-6H-pyrido[4,3 b]carbazole. These data, and previous work from our laboratory, show that the drug-complexed ligand is sorted to lysosomes whereas the receptor is recycled to the plasma membrane. The lysosomal processing of the ligand/drug construct depends on the linker between the ligand sequence and the cytotoxic moiety. We show that heptagastrin linked to ellipticine via a succinoyl-substituted pentapeptide, AlaLeuAlaLeuAla, is at least 10(3) more toxic to cholecystokinin type B receptor-positive NIH/3T3 cells than to isogenic NIH/3T3 cells lacking the receptor. The conjugated drug eradicated all receptor-positive tumor cells in vivo without producing any general toxicity. The data indicate that the density of the cell surface receptor, the properties of the cytotoxic moiety, and the correct processing of the drug-conjugated ligand in lysosomes are crucial to the effectiveness of a receptor-targeted drug. PMID- 9751699 TI - Modular synthesis of de novo-designed metalloproteins for light-induced electron transfer. AB - The design and chemical synthesis of two de novo four-helix bundle proteins is described; each protein has two bound cofactors. Their construction from purified peptides is based on the modular assembly of different amphiphilic helices by chemoselective coupling to a cyclic peptide template. In the hydrophobic interior of the antiparallel four-helix bundle these proteins contain a heme in a binding pocket with two ligating histidine residues. A ruthenium-tris(bipyridine) complex is covalently bound to different positions at the hydrophilic side of one of the heme-binding helices. Laser-induced electron transfer across the varied distance through this helix has been studied and compared with a pathway analysis. The UV visible, CD, and mass spectra are consistent with the structure and orientation predetermined by the template. PMID- 9751700 TI - Counting individual sulfur atoms in a protein by ultrahigh-resolution Fourier transform ion cyclotron resonance mass spectrometry: experimental resolution of isotopic fine structure in proteins. AB - A typical molecular ion mass spectrum consists of a sum of signals from species of various possible isotopic compositions. Only the monoisotopic peak (e.g., all carbons are 12C; all nitrogens are 14N, etc.) has a unique elemental composition. Every other isotope peak at approximately integer multiples of approximately 1 Da higher in nominal mass represents a sum of contributions from isotope combinations differing by a few mDa (e.g., two 13C vs. two 15N vs. one 13C and one 15N vs. 34S, vs. 18O, etc., at approximately 2 Da higher in mass than the monoisotopic mass). At sufficiently high mass resolving power, each of these nominal-mass peaks resolves into its isotopic fine structure. Here, we report resolution of the isotopic fine structure of proteins up to 15.8 kDa (isotopic 13C,15N doubly depleted tumor suppressor protein, p16), made possible by electrospray ionization followed by ultrahigh-resolution Fourier transform ion cyclotron resonance mass analysis at 9.4 tesla. Further, a resolving power of m/Deltam50% approximately 8,000,000 has been achieved on bovine ubiquitin (8.6 kDa). These results represent a 10-fold increase in the highest mass at which isotopic fine structure previously had been observed. Finally, because isotopic fine structure reveals elemental composition directly, it can be used to confirm or determine molecular formula. For p16, for example, we were able to determine (5.1 +/- 0.3) the correct number (five) of sulfur atoms solely from the abundance ratio of the resolved 34S peak to the monoisotopic peak. PMID- 9751701 TI - Real time detection of DNA.RNA hybridization in living cells. AB - Demonstrating hybridization between an antisense oligodeoxynucleotide and its mRNA target has proven to be extremely difficult in living cells. To address this fundamental problem in antisense research, we synthesized "molecular beacon" (MB) reporter oligodeoxynucleotides with matched fluorescent donor and acceptor chromophores on their 5' and 3' ends. In the absence of a complementary nucleic acid strand, the MB remains in a stem-loop conformation where fluorescence resonance energy transfer prevents signal emission. On hybridization with a complementary sequence, the stem-loop structure opens increasing the physical distance between the donor and acceptor moieties thereby reducing fluorescence resonance energy transfer and allowing a detectable signal to be emitted when the beacon is excited by light of the appropriate wavelength. Solution hybridization studies revealed that in the presence of a complementary strand targeted MB could yield up to a 60-fold increase in fluorescence intensity in comparison to control MB. By using a fluorescence microscope fitted with UV fluoride lenses, the detection limit of preformed MB/target sequence duplexes microinjected into cells was found to be >/=1 x 10(-1) ag of MB, or approximately 10 molecules of mRNA. On the basis of this exquisite sensitivity, real-time detection of MB/target mRNA hybridization in living cells was attempted by microinjecting MB targeted to the vav protooncogene, or control MB, into K562 human leukemia cells. Within 15 min, confocal microscopy revealed fluorescence in cells injected with targeted, but not control, MB. These studies suggest that real-time visualization and localization of oligonucleotide/mRNA interactions is now possible. MB could find utility in studying RNA processing, trafficking, and folding in living cells. We hypothesize that MB may also prove useful for finding targetable mRNA sequence under physiologic conditions. PMID- 9751702 TI - Elucidation of conditions allowing conversion of penicillin G and other penicillins to deacetoxycephalosporins by resting cells and extracts of Streptomyces clavuligerus NP1. AB - Using resting cells and extracts of Streptomyces clavuligerus NP1, we have been able to convert penicillin G (benzylpenicillin) to deacetoxycephalosporin G. Conversion was achieved by increasing by 45x the concentration of FeSO4 (1.8 mM) and doubling the concentration of alpha-ketoglutarate (1.28 mM) as compared with standard conditions used for the normal cell-free conversion of penicillin N to deacetoxycephalosporin C. ATP, MgSO4, KCl, and DTT, important in cell-free expansion of penicillin N, did not play a significant role in the ring expansion of penicillin G by resting cells or cell-free extracts. When these conditions were used with 14 other penicillins, ring expansion was achieved in all cases. PMID- 9751703 TI - Inhibition of human telomerase by 2'-O-methyl-RNA. AB - Telomerase, a ribonucleoprotein up-regulated in many types of cancers, possesses an RNA template necessary to bind and extend telomere ends. The intrinsic accessibility of telomerase to incoming nucleic acids makes the RNA template an ideal target for inhibition by oligonucleotides. We report here that 2'-O-methyl RNA (2'-O-meRNA), an oligonucleotide chemistry known to exert sequence-specific effects in cell culture and animals, inhibits telomerase with potencies superior to those possessed by analogous peptide nucleic acids (PNAs). Potent inhibition relative to PNAs is surprising, because the binding affinity of 2'-O-meRNAs for complementary RNA is low relative to analogous PNAs. A 2'-O-meRNA oligomer with terminal phosphorothioate substitutions inhibits telomerase sequence-selectively within human-tumor-derived DU145 cells when delivered with cationic lipids. In contrast to the ability of 2'-O-meRNA oligomers to inhibit telomerase, the binding of a 2'-O-meRNA to an inverted repeat within plasmid DNA was not detectable, whereas binding of PNA was efficient, suggesting that the relative accessibility of the telomerase RNA template is essential for inhibition by 2'-O meRNA. Inhibition of telomerase by 2'-O-meRNA will facilitate probing the link between telomerase activity and sustained cell proliferation and may provide a basis for the development of chemopreventive and chemotherapeutic agents. PMID- 9751704 TI - RNA folding causes secondary structure rearrangement. AB - The secondary structure of the P5abc subdomain (a 56-nt RNA) of the Tetrahymena thermophila group I intron ribozyme has been determined by NMR. Its base pairing in aqueous solution in the absence of magnesium ions is significantly different from the RNA in a crystal but is consistent with thermodynamic predictions. On addition of magnesium ions, the RNA folds into a tertiary structure with greatly changed base pairing consistent with the crystal structure: three Watson-Crick base pairs, three G.U base pairs, and an extra-stable tetraloop are lost. The common assumption that RNA folds by first forming secondary structure and then forming tertiary interactions from the unpaired bases is not always correct. PMID- 9751705 TI - Characterization of covalent adriamycin-DNA adducts. AB - Adriamycin is a popular antineoplastic agent whose ability to form covalent adducts with DNA has been correlated to cellular apoptosis (programmed cell death) in tumor models. We have isolated and purified this adduct formed under oxido-reductive (Fenton) conditions in Tris buffer. We show by homo- and heteronuclear NMR spectroscopy that the covalent Adriamycin-DNA adduct is structurally equivalent to that resulting from direct reaction with formaldehyde. Covalent linkage of the drug to one of the DNA strands confers remarkable stability to the duplex, indicated by a 162-fold reduction in the rate of strand displacement compared with the complex with noncovalently bound drug. Glyceraldehyde also engenders covalent Adriamycin-DNA complexes, providing a possible relevant biological context for in vivo adduct formation. PMID- 9751707 TI - A novel glucose-responsive element in the human insulin gene functions uniquely in primary cultured islets. AB - Insulin gene transcription is limited to the beta cells within the mammalian pancreas and, like insulin secretion, is regulated by glucose. Our previous studies in primary cultured beta cells suggested the presence of a strong glucose responsive enhancer element between base pairs -341 and -260 of the human insulin promoter, the same region in which a transcriptional repressor had been identified in beta-cell tumor lines. In an attempt to map these promoter activities and resolve these conflicting data, we designed minienhancer constructs spanning this region, and tested them in primary cultured and immortalized cells. One sequence, the Z element (base pairs -292 to -243), functions as both a potent glucose-responsive transcriptional enhancer in primary cultured islet cells and as a transcriptional repressor in immortalized beta and nonbeta cells and in primary fibroblasts. In addition, the Z element binds a novel glucose-responsive protein complex that is found in the nuclei of primary cultured islet cells, but not in the nuclei of tumor cells or primary cultured fibroblasts. These data demonstrate a critical role for the Z element in human insulin gene transcription and its regulation by glucose. PMID- 9751706 TI - Proteolytic activation of PKN by caspase-3 or related protease during apoptosis. AB - PKN, a fatty acid- and Rho-activated serine/threonine kinase having a catalytic domain highly homologous to protein kinase C (PKC), was cleaved at specific sites in apoptotic Jurkat and U937 cells on Fas ligation and treatment with staurosporin or etoposide, respectively. The cleavage of PKN occurred with a time course similar to that of PKCdelta, a known caspase substrate. This proteolysis was inhibited by a caspase inhibitor, acetyl-Asp-Glu-Val-Asp-aldehyde. The cleavage fragments were generated in vitro from PKN by treatment with recombinant caspase-3. Site-directed mutagenesis of specific aspartate residues prevented the appearance of these fragments. These results indicate that PKN is cleaved by caspase-3 or related protease during apoptosis. The major proteolysis took place between the amino-terminal regulatory domain and the carboxyl-terminal catalytic domain, and it generated a constitutively active kinase fragment. The cleavage of PKN may contribute to signal transduction, eventually leading to apoptosis. PMID- 9751708 TI - Reactivity of the HIV-1 nucleocapsid protein p7 zinc finger domains from the perspective of density-functional theory. AB - The reaction of the human immunodeficiency virus type 1 (HIV-1) nucleocapsid protein p7 (NCp7) with a variety of electrophilic agents was investigated by experimental measurements of Trp37 fluorescence decay and compared with theoretical measures of reactivity based on density-functional theory in the context of the hard and soft acids and bases principle. Statistically significant correlations were found between rates of reaction and the ability of these agents to function as soft electrophiles. Notably, the molecular property that correlated strongest was the ratio of electronegativity to hardness, chi2/eta, a quantity related to the capacity of an electrophile to promote a soft (covalent) reaction. Electronic and steric determinants of the reaction were also probed by Fukui function and frontier-orbital overlap analysis in combination with protein ligand docking methods. This analysis identified selective ligand docking regions within the conserved zinc finger domains that promoted reaction. The Cys49 thiolate was found overall to be the NCp7 site most susceptible to electrophilic attack. PMID- 9751709 TI - An activity in rat tissues that modifies nitrotyrosine-containing proteins. AB - Homogenates from rat spleen and lung could modify nitrotyrosine-containing BSA. With incubation, nitrotyrosine-containing BSA lost its epitope to a monoclonal antibody that selectively recognized nitrotyrosine-containing proteins. In the presence of protease inhibitors, the loss of the nitrotyrosine epitope occurred without protein degradation and hydrolysis. This activity was found in supernatant but not particulate fractions of spleen homogenates. The factor was heat labile, was sensitive to trypsin treatment, and was retained after passage through a membrane with a 10-kDa retention. The activity was time- and protein concentration dependent. The activity increased about 2-fold in spleen extracts with endotoxin (bacterial lipopolysaccharide) treatment of animals, suggesting that the activity is inducible or regulatable. Other nitrotyrosine-containing proteins also served as substrates, while free nitrotyrosine and some endogenous nitrotyrosine-containing proteins in tissue extracts were poor substrates. Although the product and possible cofactors for this reaction have not yet been identified, this activity may be a "nitrotyrosine denitrase" that reverses protein nitration and, thus, decreases peroxynitrite toxicity. This activity was not observed in homogenates from rat liver or kidney, suggesting that there may also be some tissue specificity for the apparent denitrase activity. PMID- 9751710 TI - Transcriptional repression by AML1 and LEF-1 is mediated by the TLE/Groucho corepressors. AB - The mammalian AML/CBFalpha runt domain (RD) transcription factors regulate hematopoiesis and osteoblast differentiation. Like their Drosophila counterparts, most mammalian RD proteins terminate in a common pentapeptide, VWRPY, which serves to recruit the corepressor Groucho (Gro). Using a yeast two-hybrid assay, in vitro association and pull-down experiments, we demonstrate that Gro and its mammalian homolog TLE1 specifically interact with AML1 and AML2. In addition to the VWRPY motif, other C-terminal sequences are required for these interactions with Gro/TLE1. TLE1 inhibits AML1-dependent transactivation of the T cell receptor (TCR) enhancers alpha and beta, which contain functional AML binding sites, in transfected Jurkat T cells. LEF-1 is an additional transcription factor that mediates transactivation of TCR enhancers. LEF-1 and its Drosophila homolog Pangolin (Pan) are involved in the Wnt/Wg signaling pathway through interactions with the coactivator beta-catenin and its highly conserved fly homolog Armadillo (Arm). We show that TLE/Gro interacts with LEF-1 and Pan, and inhibits LEF-1:beta catenin-dependent transcription. These data indicate that, in addition to their activity as transcriptional activators, AML1 and LEF-1 can act, through recruitment of the corepressor TLE1, as transcriptional repressors in TCR regulation and Wnt/Wg signaling. PMID- 9751711 TI - Compartmentation of phosphoglycerate kinase in Trypanosoma brucei plays a critical role in parasite energy metabolism. AB - African trypanosomes compartmentalize glycolysis in a microbody, the glycosome. When growing in the mammalian bloodstream, trypanosomes contain only a rudimentary mitochondrion, and the first seven glycolytic enzymes, including phosphoglycerate kinase, are located in the glycosome. Procyclic trypanosomes, growing in the gut of tsetse flies, possess a fully developed mitochondrion that is active in oxidative phosphorylation. The first six glycolytic enzymes are still glycosomal, but phosphoglycerate kinase is now found in the cytosol. We demonstrate here that bloodstream trypanosomes are killed by expression of cytosolic phosphoglycerate kinase. The toxicity depends on both enzyme activity and cytosolic location. One possible explanation is that cytosolic phosphoglycerate kinase creates an ATP-generating shunt in the cytosol, thus preventing full ATP regeneration in the glycosome and ultimately inhibiting the first, ATP-consuming, steps of glycolysis. PMID- 9751712 TI - HIV-1 tat binds TAFII250 and represses TAFII250-dependent transcription of major histocompatibility class I genes. AB - HIV Tat, a transactivator of viral transcription, represses transcription of major histocompatibility (MHC) class I genes. Repression depends exclusively on the C-terminal domain of Tat, although the mechanism of this repression has not been known. We now show that repression results from the interaction of Tat with the TAFII250 component of the general transcription factor, TFIID. The C-terminal domain of Tat binds to a site on TAFII250 that overlaps the histone acetyl transferase domain, inhibiting TAFII250 histone acetyl transferase activity. Furthermore, promoters repressed by Tat, including the MHC class I promoter, are dependent on TAFII250 whereas those that are not repressed by Tat, such as SV40 and MuLV promoters, are independent of functional TAFII250. Thus, Tat repression of MHC class I transcription would be one mechanism by which HIV avoids immune surveillance. PMID- 9751714 TI - Moieties in an RNA promoter specifically recognized by a viral RNA-dependent RNA polymerase. AB - RNAs 33 nucleotides in length can direct accurate initiation of subgenomic RNA synthesis by the brome mosaic virus RNA-dependent RNA polymerase (RdRp), provided that the native sequences are maintained at five positions: -17, -14, -13, -11, and the +1 initiation site. The functional groups in the bases of these essential nucleotides required to interact with RdRp were examined by using chemically synthesized RNAs containing base analogs at each of the five positions. Analysis using a template competition assay revealed that the mode of recognition for the initiation nucleotide (+1) is distinct from that of the other essential nucleotides in the promoter. Competition experiments also determined that three template nucleotides are sufficient for stable interaction with RdRp. These results identify base moieties in the brome mosaic virus subgenomic promoter required for efficient RNA synthesis and support the hypothesis that the recognition of a RNA promoter by a viral RdRp is analogous to the recognition of DNA promoters by DNA-dependent RNA polymerases. PMID- 9751713 TI - ATP hydrolysis catalyzed by human replication factor C requires participation of multiple subunits. AB - Human replication factor C (hRFC) is a five-subunit protein complex (p140, p40, p38, p37, and p36) that acts to catalytically load proliferating cell nuclear antigen onto DNA, where it recruits DNA polymerase delta or epsilon to the primer terminus at the expense of ATP, leading to processive DNA synthesis. We have previously shown that a subcomplex of hRFC consisting of three subunits (p40, p37, and p36) contained DNA-dependent ATPase activity. However, it is not clear which subunit(s) hydrolyzes ATP, as all five subunits include potential ATP binding sites. In this report, we introduced point mutations in the putative ATP binding sequences of each hRFC subunit and examined the properties of the resulting mutant hRFC complex and the ATPase activity of the hRFC or the p40.p37.p36 complex. A mutation in any one of the ATP binding sites of the p36, p37, p40, or p140 subunits markedly reduced replication activity of the hRFC complex and the ATPase activity of the hRFC or the p40.p37.p36 complex. A mutation in the ATP binding site of the p38 subunit did not alter the replication activity of hRFC. These findings indicate that the replication activity of hRFC is dependent on efficient ATP hydrolysis contributed to by the action of four hRFC subunits. PMID- 9751715 TI - Creation of RNA molecules that recognize the oxidative lesion 7,8-dihydro-8 hydroxy-2'-deoxyguanosine (8-oxodG) in DNA. AB - We used in vitro evolution to obtain RNA molecules that specifically recognize and bind with high affinity to the oxidative lesion 7, 8-dihydro-8-hydroxy-2' deoxyguanosine (8-oxodG) in DNA. A pool of approximately 10(15) RNA molecules containing a random insert of 45 nucleotides in length was subject to 10 successive rounds of chromatographic enrichment using an 8-oxodG affinity matrix, reverse transcription, PCR amplification, and RNA synthesis. Selected RNA molecules bind to 8-oxodG located at the 3' terminus (Kd QAQB-. in bacterial reaction centers of Rhodobacter sphaeroides determined by a driving force assay. AB - The mechanism of the electron transfer reaction, QA-.QB --> QAQB-., was studied in isolated reaction centers from the photosynthetic bacterium Rhodobacter sphaeroides by replacing the native Q10 in the QA binding site with quinones having different redox potentials. These substitutions are expected to change the intrinsic electron transfer rate by changing the redox free energy (i.e., driving force) for electron transfer without affecting other events that may be associated with the electron transfer (e.g., protein dynamics or protonation). The electron transfer from QA-. to QB was measured by three independent methods: a functional assay involving cytochrome c2 to measure the rate of QA-. oxidation, optical kinetic spectroscopy to measure changes in semiquinone absorption, and kinetic near-IR spectroscopy to measure electrochromic shifts that occur in response to electron transfer. The results show that the rate of the observed electron transfer from QA-. to QB does not change as the redox free energy for electron transfer is varied over a range of 150 meV. The strong temperature dependence of the observed rate rules out the possibility that the reaction is activationless. We conclude, therefore, that the independence of the observed rate on the driving force for electron transfer is due to conformational gating, that is, the rate limiting step is a conformational change required before electron transfer. This change is proposed to be the movement, controlled kinetically either by protein dynamics or intermolecular interactions, of QB by approximately 5 A as observed in the x-ray studies of Stowell et al. [Stowell, M. H. B., McPhillips, T. M., Rees, D. C., Soltis, S. M., Abresch, E. & Feher, G. (1997) Science 276, 812-816]. PMID- 9751727 TI - v-SNARE-dependent secretion is required for phagocytosis. AB - Phagosomes are generally believed to form by gradual apposition of the plasma membrane of leukocytes onto the surface of invading microorganisms. The internalization of the encapsulated particle is therefore predicted to reduce the surface area of the phagocyte. Contrary to this prediction, we observed that phagocytosis is associated with a net increase in cell surface area, suggesting the concomitant occurrence of exocytosis. Selective cleavage of components of the secretory machinery by microinjection or transfection of bacterial neurotoxins induced a pronounced inhibition of phagocytosis. These observations indicate that vesicle-soluble N-ethylmaleimide-sensitive factor attachment protein receptor mediated exocytosis of endomembranes is essential for optimal completion of particle internalization during phagocytosis. PMID- 9751728 TI - Distinct steady-state nuclear receptor coregulator complexes exist in vivo. AB - Transcriptional regulation by members of the nuclear hormone receptor superfamily is a modular process requiring the mediation of distinct subclasses of coregulators. These subclasses include members of the steroid receptor coactivator-1 (SRC-1) coactivator family, p300/CBP and their associated proteins, such as p300/CBP-associated factor, human homologs of SWI/SNF proteins such as BRG-1, and the less well-characterized E3 ubiquitin-protein ligases E6 papillomavirus protein-associated protein and receptor-potentiating factor-1. Because functional studies indicate that these coregulators may form higher order complexes, we analyzed steady-state complexes of different coregulator subclasses in vivo. T47D and HeLa cell lysates were subjected to biochemical fractionation and screened by immunoblotting using coregulator-specific antibodies. We show that different subclasses of nuclear receptor coregulators exhibit distinct fractionation profiles. Furthermore, evidence is provided that SRC-1 family members may exist in vivo in heteromultimeric forms with each other. In addition, we demonstrate that liganded PR is present in stable complexes containing SRC-1 and transcription intermediary factor 2 (TIF2) in vivo. Our results suggest that the assembly of large, modular transcriptional complexes by recruitment of distinct subclasses of preformed coregulator subcomplexes may be involved in transcriptional regulation by activated nuclear receptors. PMID- 9751729 TI - Apoptotic proteins Reaper and Grim induce stable inactivation in voltage-gated K+ channels. AB - Drosophila genes reaper, grim, and head-involution-defective (hid) induce apoptosis in several cellular contexts. N-terminal sequences of these proteins are highly conserved and are similar to N-terminal inactivation domains of voltage-gated potassium (K+) channels. Synthetic Reaper and Grim N terminus peptides induced fast inactivation of Shaker-type K+ channels when applied to the cytoplasmic side of the channel that was qualitatively similar to the inactivation produced by other K+ channel inactivation particles. Mutations that reduce the apoptotic activity of Reaper also reduced the synthetic peptide's ability to induce channel inactivation, indicating that K+ channel inactivation correlated with apoptotic activity. Coexpression of Reaper RNA or direct injection of full length Reaper protein caused near irreversible block of the K+ channels. These results suggest that Reaper and Grim may participate in initiating apoptosis by stably blocking K+ channels. PMID- 9751730 TI - Vascular endothelial growth factor B (VEGF-B) binds to VEGF receptor-1 and regulates plasminogen activator activity in endothelial cells. AB - The vascular endothelial growth factor (VEGF) family has recently expanded by the identification and cloning of three additional members, namely VEGF-B, VEGF-C, and VEGF-D. In this study we demonstrate that VEGF-B binds selectively to VEGF receptor-1/Flt-1. This binding can be blocked by excess VEGF, indicating that the interaction sites on the receptor are at least partially overlapping. Mutating the putative VEGF receptor-1/Flt-1 binding determinants Asp63, Asp64, and Glu67 to alanine residues in VEGF-B reduced the affinity to VEGF receptor-1 but did not abolish binding. Mutational analysis of conserved cysteines contributing to VEGF B dimer formation suggest a structural conservation with VEGF and platelet derived growth factor. Proteolytic processing of the 60-kDa VEGF-B186 dimer results in a 34-kDa dimer containing the receptor-binding epitopes. The binding of VEGF-B to its receptor on endothelial cells leads to increased expression and activity of urokinase type plasminogen activator and plasminogen activator inhibitor 1, suggesting a role for VEGF-B in the regulation of extracellular matrix degradation, cell adhesion, and migration. PMID- 9751731 TI - Mitochondrial reactive oxygen species trigger hypoxia-induced transcription. AB - Transcriptional activation of erythropoietin, glycolytic enzymes, and vascular endothelial growth factor occurs during hypoxia or in response to cobalt chloride (CoCl2) in Hep3B cells. However, neither the mechanism of cellular O2 sensing nor that of cobalt is fully understood. We tested whether mitochondria act as O2 sensors during hypoxia and whether hypoxia and cobalt activate transcription by increasing generation of reactive oxygen species (ROS). Results show (i) wild type Hep3B cells increase ROS generation during hypoxia (1. 5% O2) or CoCl2 incubation, (ii) Hep3B cells depleted of mitochondrial DNA (rho0 cells) fail to respire, fail to activate mRNA for erythropoietin, glycolytic enzymes, or vascular endothelial growth factor during hypoxia, and fail to increase ROS generation during hypoxia; (iii) rho0 cells increase ROS generation in response to CoCl2 and retain the ability to induce expression of these genes; and (iv) the antioxidants pyrrolidine dithiocarbamate and ebselen abolish transcriptional activation of these genes during hypoxia or CoCl2 in wild-type cells, and abolish the response to CoCl2 in rho degrees cells. Thus, hypoxia activates transcription via a mitochondria-dependent signaling process involving increased ROS, whereas CoCl2 activates transcription by stimulating ROS generation via a mitochondria independent mechanism. PMID- 9751732 TI - Isolation of yeast mutants defective for localization of vacuolar vital dyes. AB - An application of flow cytometric sorting is used for isolation of Saccharomyces cerevisiae mutants that mislocalize vacuolar vital dyes. This screen is based on the ability of a lipophilic styryl compound, N-(3-triethylammoniumpropyl)-4-(6-(4 (diethylamino)phenyl)hexatrie nyl )pyridinium dibromide (FM4-64), to label endocytic intermediates from the plasma membrane to the vacuole membrane at 15 degreesC. Cells stained at 15 degreesC for both FM4-64 and carboxydichlorofluorescein diacetate (a vacuolar luminal vital stain), had a pronounced shift in red/green fluorescence from cells stained at 30 degrees or 38 degreesC. Flow cytometric selection based on this characteristic shift allowed the isolation of 16 mutants. These comprised 12 complementation groups, which we have designated SVL for styryl dye vacuolar localization. These groups were put into three classes. Class I mutants contain very large vacuoles; class II mutants have very fragmented vacuoles; and class III mutants show the strongest svl phenotype with punctate/diffuse FM4-64 staining. Limited genetic overlap was observed with previously isolated mutants, namely svl2/vps41, svl6/vps16, and svl7/fab1. The remaining svl mutants appear to represent novel genes, two of which showed temperature-sensitive vacuole staining morphology. Another mutant, svl8, displayed defects in uptake and sorting of phosphatidylcholine and phosphatidylethanolamine. Our flow cytometric strategy may be useful for isolation of other mutants where mislocalization of fluorescent compounds can be detected. PMID- 9751733 TI - Evidence that the nucleotide exchange and hydrolysis cycle of G proteins causes acute desensitization of G-protein gated inward rectifier K+ channels. AB - The G-protein gated inward rectifier K+ channel (GIRK) is activated in vivo by the Gbeta gamma subunits liberated upon Gi-coupled receptor activation. We have recapitulated the acute desensitization of receptor-activated GIRK currents in heterologous systems and shown that it is a membrane-delimited process. Its kinetics depends on the guanine nucleotide species available and could be accounted for by the nucleotide exchange and hydrolysis cycle of G proteins. Indeed, acute desensitization is abolished by nonhydrolyzable GTP analogues. Whereas regulators of G-protein signaling (RGS) proteins by their GTPase activating protein activities are regarded as negative regulators, a positive regulatory function of RGS4 is uncovered in our study; the opposing effects allow RGS4 to potentiate acute desensitization without compromising GIRK activation. PMID- 9751735 TI - Habitat structure determines competition intensity and invasion success in gecko lizards. AB - Species diversity is correlated with structural complexity in many animal communities; however, experimental tests of the mechanisms underlying this important relationship are rare, especially in terrestrial communities. We manipulated physical features of the habitat of gecko lizards and measured the effect on exploitation competition for insects. Increasing both the dispersion of food resources and microhabitat topography dramatically reduced interspecific competition. Adding topographic structure reduced the advantages of the larger, faster, invasive species. Interindividual spacing decreased, but intraspecific agonistic interference increased in the more territorial, resident species. Human structural alterations of the environment facilitate invasion and competitive displacement in this system. Physical microhabitat structure can potentially affect species interactions through a variety of complex mechanisms. PMID- 9751734 TI - The relationships between notochord and floor plate in vertebrate development revisited. AB - By using the quail-chicken chimera system, we have previously shown that during development of the spinal cord, floor plate cells are inserted between neural progenitors giving rise to the alar plates. These cells are derived from the regressing Hensen's node or cordoneural hinge (HN-CNH). This common population of HN-CNH cells gives rise to three types of midline descendants: notochord, floor plate, and dorsal endoderm. Here we find that HNF3beta, an important gene in the development of the midline structures, is continuously expressed in the HN-CNH cells and their derivatives, floor plate, notochord, and dorsal endoderm. Experiments in which the notochord was removed in the posterior region of either normal chicken or of quail-chicken chimeras in which a quail HN had been grafted showed that the floor plate develops in a cell-autonomous manner in the absence of notochord. Absence of floor plate observed at the posterior level of the excision results from removal of HN-CNH material, including the future floor plate, and not from the lack of an inductive signal of notochord origin. PMID- 9751736 TI - The common marmoset: a new world primate species with limited Mhc class II variability. AB - The common marmoset (Callithrix jacchus) is a New World primate species that is highly susceptible to fatal infections caused by various strains of bacteria. We present here a first step in the molecular characterization of the common marmoset's Mhc class II genes by nucleotide sequence analysis of the polymorphic exon 2 segments. For this study, genetic material was obtained from animals bred in captivity as well as in the wild. The results demonstrate that the common marmoset has, like other primates, apparently functional Mhc-DR and -DQ regions, but the Mhc-DP region has been inactivated. At the -DR and -DQ loci, only a limited number of lineages were detected. On the basis of the number of alleles found, the -DQA and -B loci appear to be oligomorphic, whereas only a moderate degree of polymorphism was observed for two of three Mhc-DRB loci. The contact residues in the peptide-binding site of the Caja-DRB1*03 lineage members are highly conserved, whereas the -DRB*W16 lineage members show more divergence in that respect. The latter locus encodes five oligomorphic lineages whose members are not observed in any other primate species studied, suggesting rapid evolution, as illustrated by frequent exchange of polymorphic motifs. All common marmosets tested were found to share one monomorphic type of Caja-DRB*W12 allele probably encoded by a separate locus. Common marmosets apparently lack haplotype polymorphism because the number of Caja-DRB loci present per haplotype appears to be constant. Despite this, however, an unexpectedly high number of allelic combinations are observed at the haplotypic level, suggesting that Caja-DRB alleles are exchanged frequently between chromosomes by recombination, promoting an optimal distribution of limited Mhc polymorphisms among individuals of a given population. This peculiar genetic make up, in combination with the limited variability of the major histocompatability complex class II repertoire, may contribute to the common marmoset's susceptibility to particular bacterial infections. PMID- 9751738 TI - Rapid hybrid speciation in wild sunflowers. AB - Hybrid or "recombinational" speciation refers to the origin of a new homoploid species via hybridization between chromosomally or genetically divergent parental species. Theory predicts that this mode of speciation is punctuated, but there has been little empirical evidence to support this claim. Here, we test the hypothesis of rapid hybrid speciation by estimating the sizes of parental species chromosomal blocks in Helianthus anomalus, a wild sunflower species derived via hybridization between H. annuus and H. petiolaris. Analysis of the frequency spectrum of parental species chromosomal blocks with respect to predictions based on R. A. Fisher's [Fisher, R. A. (1953) Heredity 8, 187-197] junctions approach, suggests that H. anomalus arose rapidly, probably in fewer than 60 generations. This result is corroborated by independent lines of evidence demonstrating (i) a significant concordance between the genomes of H. anomalus and early generation H. annuus x H. petiolaris synthetic hybrids, and (ii) a rapid recovery of pollen fertility in these synthetic hybrid lineages. These results are not only consistent with theory but also provide a new and general method for estimating the tempo of hybrid speciation and dating the origin of hybrid zones. PMID- 9751737 TI - A mutation in human CMP-sialic acid hydroxylase occurred after the Homo-Pan divergence. AB - Sialic acids are important cell-surface molecules of animals in the deuterostome lineage. Although humans do not express easily detectable amounts of N glycolylneuraminic acid (Neu5Gc, a hydroxylated form of the common sialic acid N acetylneuraminic acid, Neu5Ac), it is a major component in great ape tissues, except in the brain. This difference correlates with lack of the hydroxylase activity that converts CMP-Neu5Ac to CMP-Neu5Gc. Here we report cloning of human and chimpanzee hydroxylase cDNAs. Although this chimpanzee cDNA is similar to the murine homologue, the human cDNA contains a 92-bp deletion resulting in a frameshift mutation. The isolated human gene also shows evidence for this deletion. Genomic PCR analysis indicates that this deletion does not occur in any of the African great apes. The gene is localized to 6p22-p23 in both humans and great apes, which does not correspond to known chromosomal rearrangements that occurred during hominoid evolution. Thus, the lineage leading to modern humans suffered a mutation sometime after the common ancestor with the chimpanzee and bonobo, potentially affecting recognition by a variety of endogenous and exogenous sialic acid-binding lectins. Also, the expression of Neu5Gc previously reported in human fetuses and tumors as well as the traces detected in some normal adult humans must be mediated by an alternate pathway. PMID- 9751739 TI - Genetic relationship of populations in China. AB - Despite the fact that the continuity of morphology of fossil specimens of modern humans found in China has repeatedly challenged the Out-of-Africa hypothesis, Chinese populations are underrepresented in genetic studies. Genetic profiles of 28 populations sampled in China supported the distinction between southern and northern populations, while the latter are biphyletic. Linguistic boundaries are often transgressed across language families studied, reflecting substantial gene flow between populations. Nevertheless, genetic evidence does not support an independent origin of Homo sapiens in China. The phylogeny also suggested that it is more likely that ancestors of the populations currently residing in East Asia entered from Southeast Asia. PMID- 9751740 TI - Amitochondriate amoebae and the evolution of DNA-dependent RNA polymerase II. AB - Unlike parasitic protist groups that are defined by the absence of mitochondria, the Pelobiontida is composed mostly of free-living species. Because of the presence of ultrastructural and cellular features that set them apart from all other eukaryotic organisms, it has been suggested that pelobionts are primitively amitochondriate and may represent the earliest-evolved lineage of extant protists. Analyses of rRNA genes, however, have suggested that the group arose well after the diversification of the earliest-evolved protists. Here we report the sequence of the gene encoding the largest subunit of DNA-dependent RNA polymerase II (RPB1) from the pelobiont Mastigamoeba invertens. Sequences within RPB1 encompass several of the conserved catalytic domains that are common to eubacterial, archaeal, and eukaryotic nuclear-encoded RNA polymerases. In RNA polymerase II, these domains catalyze the transcription of all nuclear pre-mRNAs, as well as the majority of small nuclear RNAs. In contrast with rDNA-based trees, phylogenetic analyses of RPB1 sequences indicate that Mastigamoeba represents an early branch of eukaryotic evolution. Unlike sequences from parasitic amitochondriate protists that were included in our study, there is no indication that Mastigamoeba RPB1 is attracted to the base of the eukaryotic tree artifactually. In addition, the presence of introns and a heptapeptide C-terminal repeat in the Mastigamoeba RPB1 sequence, features that are typically associated with more recently derived eukaryotic groups, raise provocative questions regarding models of protist evolution that depend almost exclusively on rDNA sequence analyses. PMID- 9751741 TI - An ion channel of the degenerin/epithelial sodium channel superfamily controls the defecation rhythm in Caenorhabditis elegans. AB - Ultradian rhythms are widespread phenomena found in various biological organisms. A typical example is the defecation behavior of the nematode Caenorhabditis elegans, which repeats at about 45-sec intervals. To elucidate the mechanism, we studied flr-1 mutants, which show very short defecation cycle periods. The mutations also affect some food-related functions, including growth rate, the expulsion step of defecation behavior, and the regulation of the dauer larva (a nonfeeding, special third-stage larva) formation in the unc-3 (Olf-1/EBF homolog) background. The flr-1 gene encodes a novel ion channel belonging to the DEG/ENaC (C. elegans degenerin and mammalian epithelial sodium channel) superfamily. A flr 1::GFP (green fluorescent protein) fusion gene that can rescue the flr-1 mutant phenotypes is expressed only in the intestine from embryos to adults. These results suggest that FLR-1 may be a component of an intestinal regulatory system that controls the defecation rhythm as well as other functions. PMID- 9751742 TI - Copy number control of a transposable element, the I factor, a LINE-like element in Drosophila. AB - The I factor is a LINE-like transposable element in Drosophila. Most strains of Drosophila melanogaster, inducer strains, contain 10-15 copies of the I factor per haploid genome located in the euchromatic regions of the chromosome arms. These are not present in a few strains known as reactive strains. I factors transpose at low frequency in inducer strains but at high frequency in the female progeny of crosses between reactive and inducer flies. We have found that the activity of the I factor promoter is sensitive to the number of copies of the first 186 nucleotides of the I factor sequence, which constitutes the 5' untranslated region. The activity of the I factor decreases as the copy number of this sequence increases. PMID- 9751743 TI - Chromosome translocation based on illegitimate recombination in human tumors. AB - Recurrent chromosome translocations in nonhematological tumors are restricted to specific subtypes, and their mechanism is currently unknown. Analysis of the sequence data of 113 interchromosomal junctions derived from 77 Ewing's tumors carrying the characteristic t(11;22) translocation indicate that, in this tumor, translocations are initiated independently on each chromosome in regions that lack site specific recombination signal. Local sequence duplications, deletions, and, most importantly, inversions that are diagnostic of DNA hairpin formation indicate that, at the breakpoint, single-stranded DNA ends are processed individually before interchromosomal joining. Taken together, these observations suggest that chromosome translocations in Ewing's tumors are mediated through a genuine illegitimate recombination mechanism. PMID- 9751744 TI - Genetic analysis of a mammalian wound-healing trait. AB - Wound healing of mammalian tissue is an essential process in the maintenance of body integrity. The general mechanism of wound healing usually studied in adult mammals is repair, in contrast to the regeneration seen in more primitive vertebrates. We recently have discovered that MRL/MpJ mice, unlike all other strains of mice tested, undergo rapid and complete wound closure that resembles regeneration. Specifically, through-and-through surgical ear hole wounds close without scarring in <4 weeks with normal gross and microanatomic architecture, including chondrogenesis. We also demonstrated that this healing is a heritable trait in inbred mice. In this study, we present results pertaining to its genetic control in progeny segregating for this phenotype. To identify the genetic loci that control the wound closure process, a genome-wide scan was performed on (MRL/MpJ-Faslpr x C57BL/6)F2 and backcross populations. In the primary screens of these populations, quantitative trait loci that control the extent of wound closure were detected on chromosomes 8, 12, and 15 and at two separate locations on chromosome 13. Evidence of further genetic control of healing was found on chromosome 7. All alleles that contribute to full wound closure are derived from the MRL/MpJ-Faslpr parent except for the quantitative trait locus on chromosome 8, which is derived from C57BL/6. PMID- 9751745 TI - The gene responsible for pseudohypoparathyroidism type Ib is paternally imprinted and maps in four unrelated kindreds to chromosome 20q13.3. AB - Hypocalcemia and hyperphosphatemia caused by parathyroid hormone (PTH)-resistance are the only discernible abnormalities in pseudohypoparathyroidism type Ib (PHP Ib). Because mutations in the PTH/PTH-related peptide receptor, a plausible candidate gene, had been excluded previously, we conducted a genome-wide search with four PHP-Ib kindreds and established linkage to a small telomeric region on chromosome 20q, which contains the stimulatory G protein gene. We, furthermore, showed that the genetic defect is imprinted paternally and thus is inherited in the same mode as the PTH-resistant hypocalcemia in kindreds with PHP-Ia and/or pseudo-pseudohypoparathyroidism, two related disorders caused by different stimulatory G protein mutations. PMID- 9751746 TI - Structural analysis of the nurse shark (new) antigen receptor (NAR): molecular convergence of NAR and unusual mammalian immunoglobulins. AB - We recently have identified an antigen receptor in sharks called NAR (new or nurse shark antigen receptor) that is secreted by splenocytes but does not associate with Ig light (L) chains. The NAR variable (V) region undergoes high levels of somatic mutation and is equally divergent from both Ig and T cell receptors (TCR). Here we show by electron microscopy that NAR V regions, unlike those of conventional Ig and TCR, do not form dimers but rather are independent, flexible domains. This unusual feature is analogous to bona fide camelid IgG in which modifications of Ig heavy chain V (VH) sequences prevent dimer formation with L chains. NAR also displays a uniquely flexible constant (C) region. Sequence analysis and modeling show that there are only two types of expressed NAR genes, each having different combinations of noncanonical cysteine (Cys) residues in the V domains that likely form disulfide bonds to stabilize the single antigen-recognition unit. In one NAR class, rearrangement events result in mature genes encoding an even number of Cys (two or four) in complementarity determining region 3 (CDR3), which is analogous to Cys codon expression in an unusual human diversity (D) segment family. The NAR CDR3 Cys generally are encoded by preferred reading frames of rearranging D segments, providing a clear design for use of preferred reading frame in antigen receptor D regions. These unusual characteristics shared by NAR and unconventional mammalian Ig are most likely the result of convergent evolution at the molecular level. PMID- 9751747 TI - LCK-phosphorylated human killer cell-inhibitory receptors recruit and activate phosphatidylinositol 3-kinase. AB - HLA-specific killer cell inhibitory receptors (KIR) are thought to impede natural killer (NK) and T cell activation programs through recruitment of the SH2 domain containing tyrosine phosphatases, SHP-1 and SHP-2, to their cytoplasmic tails (CYT). To identify other SH2 domain-containing proteins that bind KIR CYT, we used the recently described yeast two-bait interaction trap and a modified version of this system, both of which permit tyrosine phosphorylation of bait proteins. Using these systems, we show that KIR CYT, once phosphorylated by the src-family tyrosine kinase LCK, additionally bind the p85alpha regulatory subunit of phosphatidylinositol (PI) 3-kinase. Furthermore, we show that in an NK cell line, NK3.3, cross-linking of KIR results in recruitment of p85alpha to KIR and activation of PI 3-kinase lipid kinase activity. One consequence of KIR coupling to PI 3-kinase is downstream activation of the antiapoptotic protein kinase AKT. Therefore, in addition to providing negative signals, KIR may also contribute positive signals for NK and T cell growth and/or survival. PMID- 9751748 TI - BCL-6 mutations in normal germinal center B cells: evidence of somatic hypermutation acting outside Ig loci. AB - The molecular mechanism involved in the process of antigen-driven somatic hypermutation of Ig genes is unknown, but it is commonly believed that this mechanism is restricted to the Ig loci. B cell lymphomas commonly display multiple somatic mutations clustering in the 5'-regulatory region of BCL-6, a proto-oncogene encoding for a POZ/Zinc finger transcriptional repressor expressed in germinal center (GC) B cells and required for GC formation. To determine whether BCL-6 mutations represent a tumor-associated phenomenon or reflect a physiologic mechanism, we screened single human tonsillar GC B cells for mutations occurring in the BCL-6 5'-noncoding region and in the Ig variable heavy chain sequences. Thirty percent of GC B cells, but not naive B cells, displayed mutations in the 742 bp region analyzed within the first intron of BCL-6 (overall frequency: 5 x 10(-4)/bp). Accordingly, an expanded survey in lymphoid malignancies showed that BCL-6 mutations are restricted to B cell tumors displaying GC or post-GC phenotype and carrying mutated Ig variable heavy chain sequences. These results indicate that the somatic hypermutation mechanism active in GC B cells physiologically targets non-Ig sequences. PMID- 9751749 TI - Interdependence of cortical thymic epithelial cell differentiation and T-lineage commitment. AB - Thymocyte and thymic epithelial cell (TEC) development are interdependent processes. Although lineage relationships among progressively maturing thymocyte subsets have been characterized, the developmental relationships among TEC subsets are obscure. Because epithelial cells express distinct keratin (K) species as a function of differentiation stage and proliferative status, we used K expression patterns to identify mouse TEC subsets and determine their lineage relationships. As expected, cortical and medullary TEC subsets express distinct K expression patterns in the normal thymus. However, we detected two distinct cortical TEC subsets, a major K8(+)K5(-) subset and a minor K8(+)K5(+) subset, which is highly represented at the cortico-medullary junction. Both cortical TEC subsets are also present in recombination activating gene 1 (RAG-1(-/-)) and TCRbetaxdelta-/- thymi in which T-cell development is blocked at the CD4(-)CD8( )CD25(+)CD44(-) pre-T cell stage. In contrast, K8(+)K5(+) TECs predominate in the thymi of human CD3epsilon transgenic mice in which thymocyte development is blocked at an earlier CD4(-)CD8(-)CD25(-)CD44(+) stage. Transplantation of newborn human CD3epsilon transgenic thymi under the kidney capsule of RAG-1(-/-) mice results in the emergence of K8(+)K5(-) TECs concomitant with the appearance of CD25(+) thymocytes. Together, the data suggest that cortical TEC development proceeds from a K8(+)K5(+) precursor subset to a K8(+)K5(-) stage in a differentiation process concomitant with T-cell lineage commitment. PMID- 9751750 TI - Expression and crystallization of the complex of HLA-DR2 (DRA, DRB1*1501) and an immunodominant peptide of human myelin basic protein. AB - HLA-DR2 is associated with susceptibility to multiple sclerosis (MS). A peptide from human myelin basic protein (MBP, residues 85-99) was previously found to bind to purified HLA-DR2 (DRA, DRB1*1501) and to be recognized by human MBP specific T cell clones. Soluble HLA-DR2 was expressed in the baculovirus system by replacing the hydrophobic transmembrane regions and cytoplasmic segments of DRalpha and DRbeta with leucine zipper dimerization domains from the transcription factors Fos and Jun. In the expression construct, the MBP(85-99) sequence was covalently linked to the N terminus of the mature DRbeta chain. The recombinant protein was secreted by Sf9 cells infected with the recombinant baculovirus and purified by affinity chromatography. The leucine zipper dimerization domains were then cleaved from the assembled HLA-DR2/MBP peptide complex with V8 protease, and the protein was further purified by anion-exchange HPLC. Analysis by HPLC gel filtration indicated that the HLA-DR2/MBP peptide complex did not have a tendency to aggregate. The purified HLA-DR2/MBP peptide complex readily crystallized by the hanging drop method in 15-18% polyethylene glycol 6000/100 mM glycine, pH 3.5. At a synchrotron radiation source, a crystal with a tetragonal space group diffracted to a resolution of 2.6 A. The expression of such homogenous HLA-DR/peptide complexes may facilitate cocrystallization with T cell receptors as well as other molecules involved in T cell receptor recognition and signaling. PMID- 9751751 TI - Rapid deletion of rearranged T cell antigen receptor (TCR) Valpha-Jalpha segment by secondary rearrangement in the thymus: role of continuous rearrangement of TCR alpha chain gene and positive selection in the T cell repertoire formation. AB - A rearranged T cell receptor (TCR) Valpha and Jalpha gene from a cytochrome c specific T cell hybridoma was introduced into the genomic Jalpha region. The introduced TCR alpha chain gene is expressed in a majority of CD3 positive and CD4 CD8 double-negative immature thymocytes. However, only a few percent of the double-positive and single-positive thymocytes express this TCR alpha chain. This decrease is caused by a rearrangement of TCR alpha chain locus, which deletes the introduced TCR gene. Analysis of the mice carrying the introduced TCR alpha chain and the transgenic TCR beta chain from the original cytochrome c-specific T cell hybridoma revealed that positive selection efficiently rescues double-positive thymocytes from the loss of the introduced TCR alpha chain gene. In the mice with negatively selecting conditions, T cells expressing the introduced TCR alphabeta chains were deleted at the double-positive stage. However, a large number of thymocytes escape negative selection by using an endogenous TCR alpha chain created by secondary rearrangement maintaining normal thymocyte development. These results suggest that secondary rearrangements of the TCR alpha chain gene play an important role in the formation of the T cell repertoire. PMID- 9751752 TI - Trans-chromosomal recombination within the Ig heavy chain switch region in B lymphocytes. AB - Somatic DNA rearrangements in B lymphocytes, including V(D)J gene rearrangements and isotype switching, generally occur in cis, i. e., intrachromosomally. We showed previously, however, that 3 to 7% of IgA heavy chains have the VH and Calpha regions encoded in trans. To determine whether the trans-association of VH and Calpha occurred by trans-chromosomal recombination, by trans-splicing, or by trans-chromosomal gene conversion, we generated and analyzed eight IgA-secreting rabbit hybridomas with trans-associated VH and Calpha heavy chains. By ELISA and by nucleotide sequence analysis we found that the VH and Calpha regions were encoded by genes that were in trans in the germline. We cloned the rearranged VDJ Calpha gene from a fosmid library of one hybridoma and found that the expressed VH and Calpha genes were juxtaposed. Moreover, the juxtaposed VH and Calpha genes originated from different IgH alleles. From the same hybridoma, we also identified a fosmid clone with the other expected product of a trans-chromosomal recombination. The recombination breakpoint occurred within the Smicro/Salpha region, indicating that the trans-association of VH and Calpha genes occurred by trans-chromosomal recombination during isotype switching. We conclude that trans chromosomal recombination occurs at an unexpectedly high frequency (7%) within the IgH locus of B lymphocytes in normal animals, which may explain the high incidence of B-cell tumors that arise from oncogene translocation into the IgH locus. PMID- 9751753 TI - Parathyroid hormone-related protein is required for tooth eruption. AB - Parathyroid hormone (PTH)-related protein (PTHrP)-knockout mice die at birth with a chondrodystrophic phenotype characterized by premature chondrocyte differentiation and accelerated bone formation, whereas overexpression of PTHrP in the chondrocytes of transgenic mice produces a delay in chondrocyte maturation and endochondral ossification. Replacement of PTHrP expression in the chondrocytes of PTHrP-knockout mice using a procollagen II-driven transgene results in the correction of the lethal skeletal abnormalities and generates animals that are effectively PTHrP-null in all sites other than cartilage. These rescued PTHrP-knockout mice survive to at least 6 months of age but are small in stature and display a number of developmental defects, including cranial chondrodystrophy and a failure of tooth eruption. Teeth appear to develop normally but become trapped by the surrounding bone and undergo progressive impaction. Localization of PTHrP mRNA during normal tooth development by in situ hybridization reveals increasing levels of expression in the enamel epithelium before the formation of the eruption pathway. The type I PTH/PTHrP receptor is expressed in both the adjacent dental mesenchyme and in the alveolar bone. The replacement of PTHrP expression in the enamel epithelium with a keratin 14-driven transgene corrects the defect in bone resorption and restores the normal program of tooth eruption. PTHrP therefore represents an essential signal in the formation of the eruption pathway. PMID- 9751754 TI - Abnormal regulation of the leptin gene in the pathogenesis of obesity. AB - A subset of obese humans has relatively low plasma levels of leptin. This finding has suggested that in some cases abnormal regulation of the leptin gene in adipose tissue is etiologic in the pathogenesis of the obese state. The possibility that a relative decrease in leptin production can lead to obesity was tested by mating animals carrying a weakly expressed adipocyte specific aP2-human leptin transgene to C57BL/6J ob/ob mice (which do not express leptin). The transgene does not contain the regulatory elements of the leptin gene and is analogous to a circumstance in which the cis elements and/or trans factors regulating leptin RNA production are abnormal. The ob/ob mice carrying the transgene had a plasma leptin level of 1. 78 ng/ml, which is approximately one half that found in normal, nontransgenic mice (3.72 ng/ml, P < 0.01). The ob/ob animals expressing the leptin transgene were markedly obese though not as obese as ob/ob mice without the transgene. The infertility as well as several of the endocrine abnormalities generally evident in ob/ob mice were normalized in the ob/ob transgenic mice. However, the ob/ob transgenic mice had an abnormal response when placed at an ambient temperature of 4 degreesC, suggesting that different thresholds exist for the different biologic effects of leptin. Leptin treatment of the ob/ob transgenic mice resulted in marked weight loss with efficacy similar to that seen after treatment of wild-type mice. In aggregate these data suggest that dysregulation of leptin gene can result in obesity with relatively normal levels of leptin and that this form of obesity is responsive to leptin treatment. PMID- 9751755 TI - BCR/ABL-mediated leukemogenesis requires the activity of the small GTP-binding protein Rac. AB - The phenotype of hematopoietic cells transformed by the BCR/ABL oncoprotein of the Philadelphia chromosome is characterized by growth factor-independent proliferation, reduced susceptibility to apoptosis, and altered adhesion and motility. The mechanisms underlying this phenotype are not fully understood, but there is evidence that some of the properties of BCR/ABL-expressing cells are dependent on the activation of downstream effector molecules such as RAS, PI-3k, and bcl-2. We show here that the small GTP-binding protein Rac is activated by BCR/ABL in a tyrosine kinase-dependent manner. Upon transfection with a vector carrying the dominant-negative N17Rac, BCR/ABL-expressing myeloid precursor 32Dcl3 cells retained the resistance to growth factor deprivation-induced apoptosis but showed a decrease in proliferative potential in the absence of interleukin-3 (IL-3) and markedly reduced invasive properties. Moreover, compared with BCR/ABL-expressing cells, fewer BCR/ABL plus N17Rac double transfectants were capable of homing to bone marrow and spleen. Consistent with these findings, survival of SCID mice injected with the BCR/ABL plus N17Rac double transfectants was markedly prolonged as compared with that of mice injected with BCR/ABL expressing cells. Together, these data support the important role of a Rac dependent pathway(s) controlling motility in BCR/ABL-mediated leukemogenesis. PMID- 9751756 TI - The PEBP2betaMYH11 fusion created by Inv(16)(p13;q22) in myeloid leukemia impairs neutrophil maturation and contributes to granulocytic dysplasia. AB - Chromosomal translocations involving the genes encoding the alpha and beta subunits of the Pebp2/Cbf transcription factor have been associated with human acute myeloid leukemia and the preleukemic condition, myelodysplasia. Inv(16)(p13;q22) fuses the gene encoding the beta subunit of Pebp2 to the MYH11 gene encoding a smooth muscle myosin heavy chain (Smmhc). To examine the effect of the inv(16)(p13;q22) on myelopoiesis, we used the hMRP8 promoter element to generate transgenic mice expressing the Pebp2betaSmmhc chimeric fusion protein in myeloid cells. Neutrophil maturation was impaired in PEBP2betaMYH11 transgenic mice. Although the transgenic mice had normal numbers of circulating neutrophils, their bone marrow contained increased numbers of immature neutrophilic cells, which exhibited abnormal characteristics. In addition, PEBP2betaMYH11 inhibited neutrophilic differentiation in colonies derived from hematopoietic progenitors. Coexpression of both PEBP2betaMYH11 and activated NRAS induced a more severe phenotype characterized by abnormal nuclear morphology indicative of granulocytic dysplasia. These results show that PEBP2betaMYH11 can impair neutrophil development and provide evidence that alterations of Pebp2 can contribute to the genesis of myelodysplasia. PMID- 9751757 TI - Altered surfactant homeostasis and alveolar type II cell morphology in mice lacking surfactant protein D. AB - Surfactant protein D (SP-D) is one of two collectins found in the pulmonary alveolus. On the basis of homology with other collectins, potential functions for SP-D include roles in innate immunity and surfactant metabolism. The SP-D gene was disrupted in embryonic stem cells by homologous recombination to generate mice deficient in SP-D. Mice heterozygous for the mutant SP-D allele had SP-D concentrations that were approximately 50% wild type but no other obvious phenotypic abnormality. Mice totally deficient in SP-D were healthy to 7 months but had a progressive accumulation of surfactant lipids, SP-A, and SP-B in the alveolar space. By 8 weeks the alveolar phospholipid pool was 8-fold higher than wild-type littermates. There was also a 10-fold accumulation of alveolar macrophages in the null mice, and many macrophages were both multinucleated and foamy in appearance. Type II cells in the null mice were hyperplastic and contained giant lamellar bodies. These alterations in surfactant homeostasis were not associated with detectable changes in surfactant surface activity, postnatal respiratory function, or survival. The findings in the SP-D-deficient mice suggest a role for SP-D in surfactant homeostasis. PMID- 9751758 TI - Opposing actions of prostaglandins and oxytocin determine the onset of murine labor. AB - Prostaglandins (PGs) have been recently proven essential for parturition in mice. To dissect the contributions of the two cyclooxygenase (COX) isoforms to the synthesis of PGs during pregnancy, we have characterized the parturition phenotype of COX-1-deficient mice. We find that mice with targeted disruption of the COX-1 gene have delayed parturition resulting in neonatal death. Results of matings of COX-1-deficient females with COX-1 intact males, and blastocyst transfer of COX-1-deficient or -intact embryos into wild-type foster mothers, proved necessity and sufficiency of maternal COX-1 for the normal onset of labor. COX-1 expression is induced in gravid murine uterus and by in situ hybridization; this induction is localized to the decidua. Measurement of uterine PGs further confirmed that COX-1 accounted for the majority of PGF2alpha production. To evaluate the interaction of PGs with oxytocin during murine labor, we generated mice deficient in both oxytocin and COX-1. Surprisingly, the combined oxytocin and COX-1-deficient mice initiated labor at the normal time. COX-1-deficient mice demonstrated impaired luteolysis, as evidenced by elevated serum progesterone concentration and ovarian histology late in gestation, and delayed induction of uterine oxytocin receptors. In contrast, simultaneous oxytocin and COX-1 deficiency restored the normal onset of labor by allowing luteolysis in the absence of elevated PGF2alpha production. These findings demonstrate that COX-1 is essential for normal labor in the mouse, with a critical function being to overcome the luteotrophic action of oxytocin in late gestation. PMID- 9751759 TI - CC-chemokines enhance the replication of T-tropic strains of HIV-1 in CD4(+) T cells: role of signal transduction. AB - This study demonstrates that several CC-chemokines, including those that inhibit entry and replication of macrophage-tropic strains of HIV, increase the replication of T cell (T)-tropic strains in CD4(+) T cells. Enhancement of T tropic HIV replication is observed at early stages of replication, requires signaling through inhibitory guanine nucleotide-binding regulatory (Gi) proteins, and is associated with increased cell surface colocalization of CD4 and the T tropic HIV coreceptor CXCR4. These findings may further our understanding of the factors that influence the replication and spread of T-tropic strains of HIV in vivo and suggest that the use of cell signaling CC-chemokines as therapeutic agents for the purpose of limiting HIV replication in vivo should be approached with caution. PMID- 9751760 TI - Site-specific gene delivery in vivo through engineered Sendai viral envelopes. AB - Inspite of several stimulating developments in gene therapy, the formulation of a targeted gene delivery "vector" is still far from ideal. We have demonstrated the potential of reconstituted Sendai viral envelopes containing only the fusion glycoprotein (F-virosomes) in targeted delivery of reporter genes to liver cells of BALB/c mouse in vivo. The membrane fusion-mediated high efficiency of gene transfer to liver cells was ascertained following a critical evaluation of the level of the DNA, mRNA, and relevant proteins. Furthermore, the involvement of viral glycoprotein both as a unique natural ligand and as a membrane fusogen could lead to preferential transfection of parenchymal cell types of liver. The integration of transgenes in the mouse chromosomal DNA and its stable expression up to 4 mo after single i.v. administration of this gene carrier has bolstered its efficiency and novelty. Moreover, the F-virosomes did not elicit significant humoral immune response against the fusion protein in the injected animal. The findings reported here open up the possibility for considering "F-virosomes" as a promising "vehicle" for site-specific DNA delivery in gene therapy. PMID- 9751761 TI - Aberrant methylation of p16(INK4a) is an early event in lung cancer and a potential biomarker for early diagnosis. AB - The p16(INK4a) (p16) tumor suppressor gene can be inactivated by promoter region hypermethylation in many tumor types including lung cancer, the leading cause of cancer-related deaths in the U.S. We have determined the timing of this event in an animal model of lung carcinogenesis and in human squamous cell carcinomas (SCCs). In the rat, 94% of adenocarcinomas induced by the tobacco specific carcinogen 4-methylnitrosamino-1-(3-pyridyl)-1-butanone were hypermethylated at the p16 gene promoter; most important, this methylation change was frequently detected in precursor lesions to the tumors: adenomas, and hyperplastic lesions. The timing for p16 methylation was recapitulated in human SCCs where the p16 gene was coordinately methylated in 75% of carcinoma in situ lesions adjacent to SCCs harboring this change. Moreover, the frequency of this event increased during disease progression from basal cell hyperplasia (17%) to squamous metaplasia (24%) to carcinoma in situ (50%) lesions. Methylation of p16 was associated with loss of expression in both tumors and precursor lesions indicating that both alleles were functionally inactivated. The potential of using assays for aberrant p16 methylation to identify disease and/or risk was validated by detection of this change in sputum from three of seven patients with cancer and 5 of 26 cancer free individuals at high risk. These studies show for the first time that an epigenetic alteration, aberrant methylation of the p16 gene, can be an early event in lung cancer and may constitute a new biomarker for early detection and monitoring of prevention trials. PMID- 9751762 TI - Characterization of hematopoietic progenitor cells that express the transcription factor SCL, using a lacZ "knock-in" strategy. AB - Gene targeting experiments have demonstrated that the transcription factor SCL is essential for primitive and definitive hematopoiesis in the mouse. To study the functional properties of hematopoietic cells expressing SCL, we have generated mutant mice (SCLlacZ/w) in which the Escherichia coli lacZ reporter gene has been "knocked in" to the SCL locus, thereby linking beta-galactosidase expression to transcription from the SCL promoter. Bone marrow cells from heterozygous SCLlacZ/w mice were sorted into fractions expressing high, intermediate and low levels of beta-galactosidase (designated lacZhigh, lacZint, and lacZneg). Cells that were lacZhigh or lacZint were enriched for day 12 spleen colony-forming units and myeloid and erythroid colony-forming cells (CFCs). These fractions included >99% of the erythroid and >90% of the myeloid CFCs. Culture of sorted bone marrow populations on stromal cells secreting interleukin-7 or in fetal thymic organ cultures showed that B and T lymphoid progenitors were also present in the lacZhigh and lacZint fractions. These data provide a functional correlation between SCL expression and colony-forming ability in immature hematopoietic cells. Our data also suggested that expression of SCL was transient and confined to hematopoietic stem and/or progenitor cells, because the differentiated progeny of most lineages (except the erythroid) were beta galactosidase-negative. PMID- 9751763 TI - Consequences of exclusive expression in vivo of Kit-ligand lacking the major proteolytic cleavage site. AB - Membrane growth factors that are processed to produce soluble ligands may function both as soluble factors and as membrane factors. The membrane growth factor Kit-ligand (KL), the ligand of the Kit receptor tyrosine kinase, is encoded at the Sl locus, and mice carrying Sl mutations have defects in hematopoiesis, gametogenesis, and melanogenesis. Two alternatively spliced KL transcripts encode two cell-associated KL protein products, KL-1 and KL-2. The KL 2 protein lacks the major proteolytic cleavage site for the generation of soluble KL, thus representing a more stable cell-associated form of KL. We investigated the consequences of exclusive expression of KL-2 in vivo. The KL gene in embryonic stem cells was modified and KL exon 6 was replaced with a PGKneoNTRtkpA cassette by homologous recombination, and mice carrying the SlKL2 allele were obtained. SlKL2/SlKL2 mice had only slightly reduced levels of soluble KL in their serum, suggesting that in vivo KL-2 may be processed to produce soluble KL 2S. The steady-state characteristics of the hematopoietic system and progenitor numbers were normal, and the mutant animals were not anemic. However, mast cell numbers in the skin and peritoneum were reduced and the mutant animals displayed increased sensitivity to sublethal doses of gamma-irradiation. Therefore, KL-2 may substitute for KL-1 in most situations with the exception of the production of mast cells, and induced proteolytic cleavage of KL-1 to produce soluble KL may have a role in the regeneration of hematopoietic tissue after radiation injury. PMID- 9751764 TI - A strategy for genome-wide gene analysis: integrated procedure for gene identification. AB - We have developed a technique called the Integrated Procedure for Gene Identification that modifies and integrates parts from several existing techniques to increase the efficiency for genome-wide gene identification. The procedure has the following features: (i) Only the 3' portion of the expressed templates is used to ensure a match to 3' expressed sequence tag (EST) sequences; (ii) the 3' portion of the cDNA is poly dA/poly dT minus, which maintains complete representation of the expressed copies, particularly the rare copies, which otherwise would be lost heavily because of random poly dA/poly dT hybridization in the subtraction reaction; (iii) redundancy is decreased substantially by the subtraction reaction to reduce the effort for sequencing analysis; (iv) the nonsubtracted templates that largely contain the rare copies are amplified selectively with suppression PCR and are sequenced directly or through serial analysis of gene expression (SAGE); and (v) the identified sequences are matched to databases to determine whether they are cloned genes, ESTs, or novel sequences. Using this procedure in a model system, we showed that the redundant copies were largely removed, and the rates of EST matches and the novel sequence identification were significantly increased. Most of the plasmids containing the matched EST are readily available from the IMAGE consortium. This technique can be used to index genome-wide expressed genes and to identify differentially expressed genes in different cells. Compared with the existing techniques, this procedure is relatively efficient, simple, less expensive, and labor intensive. It is especially useful for standard molecular laboratories to perform genome-wide studies. PMID- 9751765 TI - Mouse embryonic stem cells carrying one or two defective Msh2 alleles respond abnormally to oxidative stress inflicted by low-level radiation. AB - Chronic oxidative stress may play a critical role in the pathogenesis of many human cancers. Here, we report that mouse embryonic stem (ES) cells deficient in DNA mismatch repair responded abnormally when exposed to low levels of ionizing radiation, a stress known to generate oxidative DNA damage. ES cells derived from mice carrying either one or two disrupted Msh2 alleles displayed an increased survival following protracted exposures to low-level ionizing radiation as compared with wild-type ES cells. The increases in survival exhibited by ES cells deficient in DNA mismatch repair appeared to have resulted from a failure to efficiently execute cell death (apoptosis) in response to radiation exposure. For each of the ES cell types, prolonged low-level radiation treatment generated oxidative genome damage that manifested as an accumulation of oxidized bases in genomic DNA. However, ES cells from Msh2(+/-) and Msh2(-/-) mice accumulated more oxidized bases as a consequence of low-level radiation exposure than ES cells from Msh2(+/+) mice. The propensity for normal cells with mismatch repair enzyme deficiencies, including cells heterozygous for inactivating mismatch repair enzyme gene mutations, to survive promutagenic genome insults accompanying oxidative stresses may contribute to the increased cancer risk characteristic of the hereditary nonpolyposis colorectal cancer syndrome. PMID- 9751766 TI - Overexpression of leptin receptors in pancreatic islets of Zucker diabetic fatty rats restores GLUT-2, glucokinase, and glucose-stimulated insulin secretion. AB - The high-Km glucose transporter, GLUT-2, and the high-Km hexokinase of beta cells, glucokinase (GK), are required for glucose-stimulated insulin secretion (GSIS). GLUT-2 expression in beta cells of Zucker diabetic fatty (ZDF) rats is profoundly reduced at the onset of beta-cell dysfunction of diabetes. Because ZDF rats are homozygous for a mutation in their leptin receptor (OB-R) gene and are therefore leptin-insensitive, we expressed the wild-type OB-R gene in diabetic islets by infusing a recombinant adenovirus (AdCMV-OB-Rb) to determine whether this reversed the abnormalities. Leptin induced a rise in phosphorylated STAT3, indicating that the transferred wild-type OB-R was functional. GLUT-2 protein rose 17-fold in AdCMV-OB-Rb-treated ZDF islets without leptin, and leptin caused no further rise. GK protein rose 7-fold without and 12-fold with leptin. Preproinsulin mRNA increased 64% without leptin and rose no further with leptin, but leptin was required to restore GSIS. Clofibrate and 9-cis-retinoic acid, the partner ligands for binding to peroxisome proliferator-activator receptor alpha (PPARalpha) and retinoid X receptor, up-regulated GLUT-2 expression in islets of normal rats, but not in ZDF rats, in which PPARalpha is very low. Because the fat content of islets of diabetic ZDF rats remains high unless they are treated with leptin, it appears that restoration of GSIS requires normalization of intracellular nutrient homeostasis, whereas up-regulation of GLUT-2 and GK is leptin-independent, requiring only high expression of OB-Rb. PMID- 9751767 TI - Strand asymmetry of CpG transitions as indicator of G1 phase-dependent origin of multiple tumorigenic p53 mutations in stem cells. AB - In dividing cells, expression of mutations is DNA strand symmetric. Of all mutations originating de novo in nondividing cells, only those in the transcribed (noncoding) strand are immediately expressed in mRNA and protein. In contrast, any new mutation in the nontranscribed (coding) strand remains unexpressed until the cells enter S phase and begin proliferation. This previously unrecognized difference enables us to examine the cell cycle-dependent origin of multiple tumorigenic mutations in stem cells. The human p53 gene, which acts as a gatekeeper in the control of G1 to S phase transition, was chosen for the analysis. Of all multiple mutations contained in p53 databases, we have tested in detail CpG transitions. Three features of CpG sites dictate this choice: C --> T transitions at methylated mCpG are the direct product of mC deamination and are replication-independent; it is easy to identify the strand bearing a primary mC - > T event because C --> T on the transcribed strand appears as G --> A on the nontranscribed strand; and CpG transitions are the most frequent (as both singular and multiple occurrences) tumor-related p53 mutations. The origin of double nonsilent CpG transitions in nondividing cells predicts a significant excess of the heterostrand (C --> T, G --> A) doublets over the homostrand (C --> T, C --> T and G --> A, G --> A) doublets. For p53, we found such an excess. Based on this result, along with the results of three other tests reported here, we conclude that the majority of multiple p53 mutations from human tumors occurred in quiescent stem cells. PMID- 9751768 TI - Effect of vitamin A supplementation on rhodopsin mutants threonine-17 --> methionine and proline-347 --> serine in transgenic mice and in cell cultures. AB - A therapeutic effect of vitamin A supplementation on the course of photoreceptor degeneration, previously reported for patients with retinitis pigmentosa, was tested in two transgenic mouse models of this disease, each carrying a dominant rhodopsin mutation. The threonine-17 --> methionine (T17M) mutation is a class II rhodopsin mutation, characterized by a thermal instability/folding defect and minimal regeneration with the chromophore. The proline-347 --> serine (P347S) mutation belongs to class I, comprised of a smaller number of mutations that exhibit no recognized biochemical abnormality in vitro. In the present study, each of the two mouse models was fed a diet containing 2.5 mg of vitamin A palmitate (control) or 102.5 mg of vitamin A palmitate (high vitamin A) per kilogram of diet. Dark-adapted, full-field electroretinograms showed that the high vitamin A diet significantly reduced the rate of decline of a-wave and b wave amplitudes in the T17M mice but had no significant effect on the decline of electroretinogram amplitude in the P347S mice. Correspondingly, histologic evaluation revealed that the treatment was associated with significantly longer photoreceptor inner and outer segments and a thicker outer nuclear layer in the T17M mice but had no effect on photoreceptor morphology in the P347S mice. In a separate series of experiments, the instability defect of the T17M mutant opsin expressed in vitro was partially alleviated by inclusion of 11-cis-retinal in the culture media. These results show that vitamin A supplementation slows the rate of photoreceptor degeneration caused by a class II rhodopsin mutation. Vitamin A supplementation may confer therapeutic benefit by stabilizing mutant opsins through increased availability of the chromophore. PMID- 9751769 TI - HIV, but not murine leukemia virus, vectors mediate high efficiency gene transfer into freshly isolated G0/G1 human hematopoietic stem cells. AB - Recent studies have opened the possibility that quiescent, G0/G1 hematopoietic stem cells (HSC) can be gene transduced; lentiviruses (such as HIV type 1, HIV) encode proteins that permit transport of the viral genome into the nucleus of nondividing cells. We and others have recently demonstrated efficient transduction by using an HIV-1-based vector gene delivery system into various human cell types including human CD34(+) cells or terminally differentiated neurons. Here we compare the transduction efficiency of two vectors, HIV-based and murine leukemia virus (MuLV)-based vectors, on untreated and highly purified human HSC subsets that are virtually all in G0/G1. The HIV vector, but not MuLV vector supernatants, transduced freshly isolated G0/G1 HSC from mobilized peripheral blood. Single-step transduction using replication-defective HIV resulted in HSC that expressed the green fluorescent protein (GFP) transgene while retaining their stem cell phenotype; clonal outgrowths of these GFP+ HSC on bone marrow stromal cells fully retained GFP expression for at least 5 weeks. MuLV-based vectors did not transduce resting HSC, as measured by transgene expression, but did so readily when the HSC were actively cycling after culture in vitro for 3 days in a cytokine cocktail. These results suggest that resting HSC may be transduced by lentiviral-based, but not MuLV, vectors and maintain their primitive phenotype, pluripotentiality, and at least in vitro, transgene expression. PMID- 9751770 TI - pRB plays an essential role in cell cycle arrest induced by DNA damage. AB - To maintain genome stability, cells with damaged DNA must arrest to allow repair of mutations before replication. Although several key components required to elicit this arrest have been discovered, much of the pathway remains elusive. Here we report that pRB acts as a central mediator of the proliferative block induced by a diverse range of DNA damaging stimuli. Rb-/- mouse embryo fibroblasts are defective in arrest after gamma-irradiation, UV irradiation, and treatment with a variety of chemotherapeutic drugs. In contrast, the pRB related proteins p107 and p130 do not play an essential part in the DNA damage response. pRB is required specifically for the G1/S phase checkpoint induced by gamma irradiation. Despite a defect in G1/S phase arrest, levels of p53 and p21 are increased normally in Rb-/- cells in response to gamma-irradiation. These results lead us to propose a model in which pRB acts as an essential downstream target of the DNA damage-induced arrest pathway. The ability of pRB to prevent replication of damaged DNA is likely to inhibit the propagation of carcinogenic mutations and may therefore contribute to its role as a tumor suppressor. Furthermore, because many cancer therapies act by damaging DNA, these findings also have implications for the treatment of tumors in which pRB is inactivated. PMID- 9751772 TI - Chemotaxis in a gliding bacterium. AB - Myxococcus xanthus cells exhibit directed motility up phosphatidylethanolamine (PE) gradients, and we suggest that PE behaves as a chemoattractant. Computer assisted stop-motion digital microscopy was used to record cell movements in slide culture. PE decreased cellular reversal frequency with molecular specificity that was correlated with the fatty acid composition. Synthetic dilauroyl (di C12:0) PE and dioleoyl (di C18:1 omega9c) PE suppressed direction reversals and stimulated movement up the gradient. Sensory adaptation occurred about 1 hr after the onset of stimulation. Null mutants in a methylated chemotaxis protein homolog (FrzCD) and a CheA/CheY homolog (FrzE) moved up a PE gradient at a reduced rate. The mutants displayed normal excitation but were defective in adaptation. A dominant, hyper-reversal mutant in the M. xanthus methyl accepting chemotaxis protein homolog, frzCD224, failed to respond to PE stimulation, which argued that PE was a transduced stimulus. Neither dilauroyl PE nor dioleoyl PE is present at high enough concentrations in vegetative or developmental PE to account for all of the chemotactic activity. It appears then that there are additional, as yet unknown, PE species that serve as autoattractants. We report on a discrete phospholipid chemoattractant in a gliding bacterium PMID- 9751771 TI - In vivo characterization of the type A and B vancomycin-resistant enterococci (VRE) VanRS two-component systems in Escherichia coli: a nonpathogenic model for studying the VRE signal transduction pathways. AB - Escherichia coli reporter strains modeling the high-level type A and B vancomycin resistances of Enterococcus faecium BM4147 and Ent. faecalis have been developed to study the respective VanR-VanS two-component regulatory systems. PvanH-, PvanRa-, PvanY-, and PvanRb-lacZ fusions report on expression from the vancomycin resistant enterococci promoters of the type A vanRSHAXYZ and type B vanRSYWHBX gene clusters. These strains also express from single-copy chromosomal genes vanRa, vanRb, or vanRSb behind their respective promoter (PvanRa or PvanRb) or vanSa or vanSb behind the rhamnose-inducible PrhaB. Results show that activation (phosphorylation) of the response regulator VanRa by its sensor kinase VanSa leads to transcriptional activation of both PvanH and PvanRa. Additionally, VanRb activates its cognate promoters PvanY and PvanRb, although this occurs only in the absence of VanSb and presumably is caused by VanRb phosphorylation by an unidentified endogenous E. coli kinase. Thus, VanSb interferes with activation of VanRb, probably by acting as a phospho-VanRb phosphatase. Although both VanRa and VanRb activate their cognate promoters, neither activates the heterologous PvanR, PvanH, or PvanY, arguing against the interchangeability of type A and B two component regulatory switches in vancomycin-resistant enterococci. VanRa also is activated by the nonpartner kinase PhoR. Because this occurs in the absence of its inducing signal (Pi limitation), PhoR autophosphorylation apparently is regulated in vivo. Furthermore, the activation of VanRa caused by cross talk from PhoR in the absence of a signal allows distinction of cross talk from crossregulation as the latter, but not the former, responds to environmental cues. PMID- 9751773 TI - Expression of the Epstein-Barr virus latent membrane protein 1 induces B cell lymphoma in transgenic mice. AB - The latent membrane protein 1 (LMP1) of the Epstein-Barr virus has transforming properties in rodent fibroblasts and is expressed in most of the cancers associated with Epstein-Barr virus (EBV) infection including posttransplant lymphomas, Hodgkin's disease, nasopharyngeal carcinoma, and AIDS-related lymphomas. In this study, three lineages of LMP1 transgenic mice were established with LMP1 expressed under the control of the Ig heavy chain promoter and enhancer. Lymphoma developed in all three lineages, and the incidence of lymphoma increased significantly with age with lymphomas developing in 42% of transgenic mice over 18 months. The expression of LMP1 was detected at high levels in the lymphoma tissues but only at trace levels in normal lymphoid tissues. Gene rearrangement of the Ig heavy chain indicated monoclonality or oligoclonality in all lymphomas, some of the lymphoid hyperplastic spleens, and some histologically normal spleens. These data reveal that LMP1, without the expression of other EBV genes, is oncogenic in vivo and indicate that LMP1 is a major contributing factor to the development of EBV-associated lymphomas. PMID- 9751774 TI - The ORF47 and ORF66 putative protein kinases of varicella-zoster virus determine tropism for human T cells and skin in the SCID-hu mouse. AB - The varicella-zoster virus (VZV) genes ORF47 and ORF66 are predicted to encode serine/threonine protein kinases, which are homologs of herpes simplex virus 1 (HSV-1) UL13, and US3. When mutants were constructed by inserting stop codons into ORF47 and ORF66, the recombinants ROka47S and ROka66S, as well as intact ROka replicated in tissue culture. In contrast, inoculation of human thymus/liver or skin implants in SCID-hu mice showed that ORF47 protein was required for viral growth in human T cells and skin. Eliminating ORF66 expression inhibited VZV infectivity for T cells partially but did not impair replication in skin compared with ROka. Infectivity for T cells and skin was restored when ROka47S virus was complemented by insertion of ORF47 into a distant, noncoding site. The ORF47 gene product is the first VZV protein identified as necessary for T cell tropism. It also is essential for skin infectivity in vivo, as is glycoprotein C. Expression of ORF66 did not compensate for the absence of the ORF47 protein. The requirement for ORF47 expression in T cells and skin indicates that this gene product, which is dispensable in vitro, has a critical role within differentiated cells that are essential targets for VZV pathogenesis in vivo. PMID- 9751775 TI - Inhibition of the p44/42 MAP kinase pathway protects hippocampal neurons in a cell-culture model of seizure activity. AB - Excessive release of glutamate and the subsequent influx of calcium are associated with a number of neurological insults that result in neuronal death. The calcium-activated intracellular signaling pathways responsible for this excitotoxic injury are largely unknown. Here, we report that PD098059, a selective inhibitor of the calcium-activated p44/42 mitogen-activated protein kinase (MAP kinase) pathway, reduces neuronal death in a cell-culture model of seizure activity. Dissociated hippocampal neurons grown chronically in the presence of kynurenate, a broad spectrum glutamate-receptor antagonist, and elevated amounts of magnesium exhibit intense seizure-like activity after the removal of these blockers of excitatory synaptic transmission. A 30-min removal of the blockers produced extensive neuronal death within 24 h as assayed by the uptake of trypan blue and the release of lactate dehydrogenase. Phospho-p44/42 MAP kinase immunoreactivity after 30 min of seizure-like activity was present in many neuronal somata and dendrites as well as some synaptic terminals, consistent with both the presynaptic and postsynaptic effects of this pathway. The addition of PD098059 (40 microM; EC50 = 10 microM) during a 30-min washout of synaptic blockers inhibited the phosphorylation of p44/42 MAP kinase and reduced both the trypan-blue staining (n = 13) and the release of lactate dehydrogenase (n = 16) by 73% +/- 18% and 75% +/- 19% (mean +/- SD), respectively. The observed neuroprotection could be caused by an effect of PD098059 on seizure-like events or on downstream signaling pathways activated by the seizure-like events. Either possibility suggests a heretofore unknown function for the p44/42 MAP kinase pathway in neurons. PMID- 9751776 TI - Direct immunogold labeling of aquaporin-4 in square arrays of astrocyte and ependymocyte plasma membranes in rat brain and spinal cord. AB - Aquaporin (AQP) water channels are abundant in the brain and spinal cord, where AQP1 and AQP4 are believed to play major roles in water metabolism and osmoregulation. Immunocytochemical analysis of the brain recently revealed that AQP4 has a highly polarized distribution, with marked expression in astrocyte end feet that surround capillaries and form the glia limitans; however, the structural organization of AQP4 has remained unknown. In freeze-fracture replicas, astrocyte end-feet contain abundant square arrays of intramembrane particles that parallel the distribution of AQP4. To determine whether astrocyte and ependymocyte square arrays contain AQP4, we employed immunogold labeling of SDS-washed freeze-fracture replicas and stereoscopic confirmation of tissue binding. Antibodies to AQP4 directly labeled approximately 33% of square arrays in astrocyte and ependymocyte plasma membranes in rat brain and spinal cord. Overall, 84% of labels were present beneath square arrays; 11% were beneath particle clusters that resembled square arrays that had been altered during fixation or cleaving; and 5% were beneath the much larger areas of glial plasma membrane that were devoid of square arrays. Based on this evidence that AQP4 is concentrated in glial square arrays, freeze-fracture methods may now provide biophysical insights regarding neuropathological states in which abnormal fluid shifts are accompanied by alterations in the aggregation state or the molecular architecture of square arrays. PMID- 9751777 TI - Cloning and localization of two multigene receptor families in goldfish olfactory epithelium. AB - Goldfish reproduction is coordinated by pheromones that are released by ovulating females and detected by males. Two highly potent pheromones, a dihydroxyprogesterone and a prostaglandin, previously have been identified, and their effects on goldfish behavior have been studied in depth. We have cloned goldfish olfactory epithelium cDNAs belonging to two multigene G-protein coupled receptor families as a step toward elucidating the molecular basis of pheromone recognition. One gene family (GFA) consists of homologs of putative odorant receptors (approximately 320 residues) found in the olfactory epithelium of other fish and mammals. The other family (GFB) consists of homologs of putative pheromone receptors found in the vomeronasal organ (VNO) of mammals and also in the nose of pufferfish. GFB receptors (approximately 840 residues) are akin to the V2R family of VNO receptors, which possess a large extracellular N-terminal domain and are homologs of calcium-sensing and metabotropic glutamate receptors. In situ hybridization showed that the two families of goldfish receptors are differentially expressed in the olfactory epithelium. GFB mRNA is abundant in rather compact cells whose nuclei are near the apical surface. In contrast, GFA mRNA is found in elongated cells whose nuclei are positioned deeper in the epithelium. Our findings support the hypothesis that the separate olfactory organ and VNO of terrestrial vertebrates arose in evolution by the segregation of distinct classes of neurons that were differentially positioned in the olfactory epithelium of a precursor aquatic vertebrate. PMID- 9751778 TI - Interpreting functional imaging studies in terms of neurotransmitter cycling. AB - Functional imaging experiments, in particular positron-emission tomography and functional magnetic resonance imaging, can be analyzed either in psychological terms or on the basis of neuroscience. In the usual psychological interpretation, stimulations are designed to activate specific mental processes identified by cognitive psychology, which are then localized by the signals in functional imaging experiments. An alternate approach would be to analyze experiments in terms of the neurobiological processes responsible for the signals. Recent in vivo 13C NMR measurements of the glutamate-to-glutamine neurotransmitter cycling in rat and human brains facilitate a neuroscientific interpretation of functional imaging data in terms of neurobiological processes since incremental neurotransmitter flux showed a 1:1 stoichiometry with the incremental rate of glucose oxidation. Because functional imaging signals depend on brain energy consumption, a quantitative relationship can be established between the signal (S) and the specific neurochemical cerebral neurotransmitter activity (N) of glutamate-to-glutamine neurotransmitter cycling. The quantitation of neuronal activity proposed has implications for the psychological design and interpretation of functional imaging experiments. Measurements of the neurotransmitter cycling flux at rest in functional imaging experiments suggest that performing cognitive tasks and sensory stimulations increases neurotransmitter cycling by only 10-20%. Therefore it cannot be assumed that reference state activities are negligible, nor that they are constant during stimulation. PMID- 9751779 TI - The effect of dynamic synapses on spatiotemporal receptive fields in visual cortex. AB - Temporal dynamics are a general feature of synaptic transmission. Recently, novel aspects of temporal dynamics of synaptic transmission have been reported in the neocortex. Here, we examine the possible effects of these dynamics on the spatiotemporal receptive fields of simple cells in V1. We do this by examining a simple model of a cortical neuron that displays stimulus orientation selectivity as a consequence of the pattern of thalamocortical synaptic weights. In our model, the receptive field structure is encoded functionally in either presynaptic probability of release or postsynaptic efficacy. We show that these different assumptions about the origin of receptive field structure lead to very different spatiotemporal dynamics in the case of flashed-bar stimulus. In addition, the results of the reverse correlation study suggest a possible test for differentiating between models. We also show that the temporal code induced by dynamic synapses can be used to distinguish between different inputs that induce the same average firing rate. PMID- 9751780 TI - Target cell-specific modulation of transmitter release at terminals from a single axon. AB - In the hippocampus, a CA3 pyramidal cell forms excitatory synapses with thousands of other pyramidal cells and inhibitory interneurons. By using sequential paired recordings from three connected cells, we show that the presynaptic properties of CA3 pyramidal cell terminals, belonging to the same axon, differ according to the type of target cell. Activation of presynaptic group III metabotropic glutamate receptors decreases transmitter release only at terminals contacting CA1 interneurons but not CA1 pyramidal cells. Furthermore, terminals contacting distinct target cells show different frequency facilitation. On the basis of these results, we conclude that the pharmacological and physiological properties of presynaptic terminals are determined, at least in part, by the target cells. PMID- 9751781 TI - Targeted disruption of the Ca2+ channel beta3 subunit reduces N- and L-type Ca2+ channel activity and alters the voltage-dependent activation of P/Q-type Ca2+ channels in neurons. AB - In comparison to the well characterized role of the principal subunit of voltage gated Ca2+ channels, the pore-forming, antagonist-binding alpha1 subunit, considerably less is understood about how beta subunits contribute to neuronal Ca2+ channel function. We studied the role of the Ca2+ channel beta3 subunit, the major Ca2+ channel beta subunit in neurons, by using a gene-targeting strategy. The beta3 deficient (beta3-/-) animals were indistinguishable from the wild type (wt) with no gross morphological or histological differences. However, in sympathetic beta3-/- neurons, the L- and N-type current was significantly reduced relative to wt. Voltage-dependent activation of P/Q-type Ca2+ channels was described by two Boltzmann components with different voltage dependence, analogous to the "reluctant" and "willing" states reported for N-type channels. The absence of the beta3 subunit was associated with a hyperpolarizing shift of the "reluctant" component of activation. Norepinephrine inhibited wt and beta3-/- neurons similarly but the voltage sensitive component was greater for N-type than P/Q-type Ca2+ channels. The reduction in the expression of N-type Ca2+ channels in the beta3-/- mice may be expected to impair Ca2+ entry and therefore synaptic transmission in these animals. This effect may be reversed, at least in part, by the increase in the proportion of P/Q channels activated at less depolarized voltage levels. PMID- 9751782 TI - Altered thalamic response to levodopa in Parkinson's patients with dopa-induced dyskinesias. AB - Parkinson's disease (PD) is a progressive neurologic condition characterized by tremor, slowness, stiffness, and unstable posture. Degeneration of dopamine producing neurons in the substantia nigra causes PD. Treatment with levodopa, a precursor of dopamine, initially ameliorates the clinical manifestations of PD. However, chronic levodopa treatment can produce severe involuntary movements (so called dopa-induced dyskinesias or DID), limiting treatment. Pallidotomy, placement of a surgical lesion in the internal segment of the globus pallidus, reduces DID. Because this result is inconsistent with current theories of both basal ganglia function and DID, it prompted us to investigate the brain's response to levodopa. We measured regional cerebral blood flow response to levodopa with positron-emission tomography in 6 PD patients with DID, 10 chronically treated PD patients without DID, 17 dopa-naive PD patients, and 11 normals. The dose of levodopa was chosen to produce clinical benefit without inducing DID. This strategy allowed us to examine the brain response to levodopa across groups without the confounding effect of differences in motor behavior. We found that the DID group had a significantly greater response in ventrolateral thalamus than the other groups. This was associated with decreased activity in primary motor cortex. These findings are consistent with increased inhibitory output from the internal segment of the globus pallidus to thalamus after levodopa administration. They provide a physiological explanation for the clinical efficacy of pallidotomy and new insights into the physiology of the basal ganglia. PMID- 9751783 TI - Specific, irreversible inactivation of the epidermal growth factor receptor and erbB2, by a new class of tyrosine kinase inhibitor. AB - A class of high-affinity inhibitors is disclosed that selectively target and irreversibly inactivate the epidermal growth factor receptor tyrosine kinase through specific, covalent modification of a cysteine residue present in the ATP binding pocket. A series of experiments employing MS, molecular modeling, site directed mutagenesis, and 14C-labeling studies in viable cells unequivocally demonstrate that these compounds selectively bind to the catalytic domain of the epidermal growth factor receptor with a 1:1 stoichiometry and alkylate Cys-773. While the compounds are essentially nonreactive in solution, they are subject to rapid nucleophilic attack by this particular amino acid when bound in the ATP pocket. The molecular orientation and positioning of the acrylamide group in these inhibitors in relation to Cys-773 entirely support these results as determined from docking experiments in a homology-built molecular model of the ATP site. Evidence is also presented to indicate that the compounds interact in an analogous fashion with erbB2 but have no activity against the other receptor tyrosine kinases or intracellular tyrosine kinases that were tested in this study. Finally, a direct comparison between 6-acrylamido-4-anilinoquinazoline and an equally potent but reversible analog shows that the irreversible inhibitor has far superior in vivo antitumor activity in a human epidermoid carcinoma xenograft model with no overt toxicity at therapeutically active doses. The activity profile for this compound is prototypical of a generation of tyrosine kinase inhibitors with great promise for therapeutic significance in the treatment of proliferative disease. PMID- 9751785 TI - An oculomotor representation area within the ventral premotor cortex. AB - We explored the ventral part of the premotor cortex (PMV) with intracortical microstimulation (ICMS) while monkeys performed a visual fixation task, to see whether the PMV is involved in oculomotor control. ICMS evoked saccades from a small-restricted region in the PMV, without evoking movements in the limbs, neck, or body. We found the saccade-evoking site in the PMV in a total of three hemispheres in two monkeys. Quantitative analysis of the effects of eye position on saccades evoked by microstimulation of the PMV characterized the evoked saccades as goal directed. The nature of the saccades evoked in the PMV contrasted with the fixed vector nature of saccades evoked by ICMS of the frontal eye field. We also found that neurons in this restricted area of the PMV were active while the animals were performing a saccade task that required them to make saccades toward targets without arm movements. These data provide evidence for the presence of an oculomotor-specific subregion within the PMV. This subregion and the surrounding skeletomotor-representing regions of the PMV seem to coordinate oculomotor and skeletomotor control in performing goal-directed motor tasks. PMID- 9751784 TI - Aminophosphinic inhibitors as transition state analogues of enkephalin-degrading enzymes: a class of central analgesics. AB - Inhibition of aminopeptidase N and neutral endopeptidase-24.11, two zinc metallopeptidases involved in the inactivation of the opioid peptides enkephalins, produces potent physiological analgesic responses, without major side-effects, in all animal models of pain in which morphine is active. Dual inhibitors of both enzymes could fill the gap between opioid analgesics and antalgics. Until now, attempts to find a compound with high affinity both for neutral endopeptidase and aminopeptidase N have failed. We report here the design of dual competitive inhibitors of both enzymes with KI values in the nanomolar range. These have been obtained by selecting R1, R2, and R3 determinants in aminophosphinic-containing inhibitors: NH2---CH(R1)P(O)---(OH)CH2---CH(R2)CONH-- CH(R3)COOH, for optimal recognition of the two enkephalin inactivating enzymes, whose active site peculiarities, determined by site-directed mutagenesis, have been taken into account. The best inhibitors were 10x more potent than described dual inhibitors in alleviating acute and inflammatory nociceptive stimuli in mice, thus providing a basis for the development of a family of analgesics devoid of opioid side effects. PMID- 9751786 TI - Alanine, not ammonia, is excreted from N2-fixing soybean nodule bacteroids. AB - Symbiotic nitrogen fixation, the process whereby nitrogen-fixing bacteria enter into associations with plants, provides the major source of nitrogen for the biosphere. Nitrogenase, a bacterial enzyme, catalyzes the reduction of atmospheric dinitrogen to ammonium. In rhizobia-leguminous plant symbioses, the current model of nitrogen transfer from the symbiotic form of the bacteria, called a bacteroid, to the plant is that nitrogenase-generated ammonia diffuses across the bacteroid membrane and is assimilated into amino acids outside of the bacteroid. We purified soybean nodule bacteroids by a procedure that removed contaminating plant proteins and found that alanine was the major nitrogen containing compound excreted. Bacteroids incubated in the presence of 15N2 excreted alanine highly enriched in 15N. The ammonium in these assays neither accumulated significantly nor was enriched in 15N. The results demonstrate that a transport mechanism rather than diffusion functions at this critical step of nitrogen transfer from the bacteroids to the plant host. Alanine may serve only as a transport species, but this would permit physiological separation of the transport of fixed nitrogen from other nitrogen metabolic functions commonly mediated through glutamate. PMID- 9751787 TI - The plant cDNA LCT1 mediates the uptake of calcium and cadmium in yeast. AB - Nonessential metal ions such as cadmium are most likely transported across plant membranes via transporters for essential cations. To identify possible pathways for Cd2+ transport we tested putative plant cation transporters for Cd2+ uptake activity by expressing cDNAs in Saccharomyces cerevisiae and found that expression of one clone, LCT1, renders the growth of yeast more sensitive to cadmium. Ion flux assays showed that Cd2+ sensitivity is correlated with an increase in Cd2+ uptake. LCT1-dependent Cd2+ uptake is saturable, lies in the high-affinity range (apparent KM for Cd2+ = 33 microM) and is sensitive to block by La3+ and Ca2+. Growth assays demonstrated a sensitivity of LCT1-expressing yeast cells to extracellular millimolar Ca2+ concentrations. LCT1-dependent increase in Ca2+ uptake correlated with the observed phenotype. Furthermore, LCT1 complements a yeast disruption mutant in the MID1 gene, a non-LCT1-homologous yeast gene encoding a membrane Ca2+ influx system required for recovery from the mating response. We conclude that LCT1 mediates the uptake of Ca2+ and Cd2+ in yeast and may therefore represent a first plant cDNA encoding a plant Ca2+ uptake or an organellar Ca2+ transport pathway in plants and may contribute to transport of the toxic metal Cd2+ across plant membranes. PMID- 9751789 TI - The absence of phloem loading in willow leaves. AB - Willow (Salix babylonica L.) is representative of a large group of plants that have extensive plasmodesmatal connections between minor vein phloem and adjoining cells. Because plasmodesmata provide a diffusion pathway for small molecules, it is unclear how sucrose could be loaded from the mesophyll into the phloem against a concentration gradient. In the studies reported here, the minor vein phloem of willow leaves plasmolyzed in approximately the same concentration of osmoticum as the mesophyll. Sucrose concentrations in mesophyll cells were greater than those reported in the literature for aphid stylet exudate from willow stems. Calculated turgor pressures in the mesophyll and minor vein phloem were greater than turgor reported in the literature for sieve elements in the stems of willow. Images of minor veins were not obtained in autoradiographs when attached leaves, or leaf pieces, were provided with 14CO2 or [14C]sucrose. Therefore, no evidence could be found for accumulation of sucrose against a concentration gradient in the minor vein phloem of willow. In these leaves, the mesophyll apparently acts as the "source" for long distance transport of sugar. The mechanism of translocation in willow, and the evolution of phloem loading, are discussed. PMID- 9751788 TI - Evidence for posttranscriptional activation of gamma-glutamylcysteine synthetase during plant stress responses. AB - Glutathione (GSH) is a key component of plant antioxidant defenses. We have sought to determine how the rate-limiting step in GSH biosynthesis, catalyzed by gamma-glutamylcysteine synthase (gammaECS) is regulated in Arabidopsis. Functional complementation of a yeast mutant deficient in this enzyme with an Arabidopsis expression library yielded two cDNAs with sequence identical to the previously described AtgammaECS. Nevertheless, the cellular concentration of GSH in these transformants was only 10% of wild-type concentrations and this was not a result of Cys availability. To explore the possibility that Arabidopsis gammaECS requires additional factors for full catalytic activity, we analyzed the GSH levels and the enzyme activities and transcript levels of both enzymes of the GSH biosynthetic pathway in Arabidopsis suspension cultures subjected to a variety of stresses that raise GSH levels. Our results demonstrate rapid posttranscriptional activation of Arabidopsis gammaECS. The implications of these findings for the mechanisms by which GSH concentrations are regulated during plant-stress responses are discussed. PMID- 9751790 TI - Neuroimaging analyses of human working memory. AB - We review a program of research that uses neuroimaging techniques to determine the functional and neural architecture of human working memory. A first set of studies indicates that verbal working memory includes a storage component, which is implemented neurally by areas in the left-hemisphere posterior parietal cortex, and a subvocal rehearsal component, which is implemented by left hemisphere speech areas, including Broca's area as well as the premotor and supplementary motor areas. We provide a number of neuroimaging dissociations between the storage and rehearsal areas. A second set of studies focuses on spatial working memory and indicates that it is mediated by a network of predominantly right-hemisphere regions that include areas in posterior parietal, occipital, and frontal cortex. We provide some suggestive evidence that these areas, too, divide into storage and rehearsal regions, with right-hemisphere posterior parietal and premotor regions subserving spatial rehearsal. In a final set of studies, we turn to "executive processes," metaprocesses that regulate the processing of working-memory contents. We focus on the executive process of inhibition as it is used in verbal working memory. We provide evidence that such inhibition is mediated by the left-hemisphere prefrontal region and that it can be dissociated from verbal storage and rehearsal processes. PMID- 9751791 TI - Quantitative analysis of sugar constituents of glycoproteins by capillary electrophoresis. AB - A method for quantitative analysis of monosaccharides including N acetylneuraminic acid derived from sialic acid-containing oligosaccharides and glycoproteins is presented. The analysis is based on the combination of chemical and enzymatic methods coupled with capillary electrophoretic (CE) separation and laser-induced fluorescence (LIF) detection. The present method utilizes a simplified acid hydrolysis procedure consisting of mild hydrolysis (0.1 M TFA) to release sialic acid and strong acid hydrolysis (2.0 N TFA) to produce amino and neutral sugars. Amino sugars released from strong acid hydrolysis of oligosaccharides and glycoproteins were reacetylated and derivatized with 8 aminopyrene-1,3,6-trisulfonate (APTS) along with neutral sugars in the presence of sodium cyanoborohydride to yield quantitatively the highly stable fluorescent APTS adducts. N-acetylneuraminic acid (Neu5Ac), a major component of most mammalian glycoproteins, was converted in a fast specific reaction by the action of neuraminic acid aldolase (N-acylneuraminate pyruvate-lyase EC 4.1.3.3) to N acetylmannosamine (ManNAc) and pyruvate. ManNAc was then derivatized with APTS in the same manner as the other monosaccharides. This method was demonstrated for the quantitation of pure Neu5Ac and the species derived from mild acid hydrolysis of 6'-sialyl-N-acetyllactosamine and bovine fetuin glycan. Quantitative recovery of the N-acetylmannosamine was obtained from a known amount of Neu5Ac in a mixture of seven other monosaccharides or from the sialylated oligosaccharides occurring in glycoproteins. The sequence of procedures consists of acid hydrolysis, enzymatic conversion and APTS derivatization which produced quantitative recovery of APTS-monosaccharide adducts. The detection limits for sugars derivatized with APTS and detected by CE-LIF are 100 pmol for Neu5Ac and 50 pmol for the other sugars. PMID- 9751792 TI - Expression of N-linked sialyl Le(x) determinants and O-glycans in the carbohydrate moiety of human amniotic fluid transferrin during pregnancy. AB - Transferrin, a glycoprotein involved in iron transport in body fluids, was isolated from amniotic fluid of a hydramniospatient by sequential anion-exchange chromatography and gel filtration. The N-glycans of human amniotic fluid transferrin (hAFT) were enzymatically liberated by PNGase-F digestion, isolated by gel filtration and fractionated by (high-pH) anion-exchange chromatography. After alkaline borohydride treatment of native hAFT, the released O-glycans were isolated by gel filtration and fractionated by anion-exchange chroma-tography. Structure elucidation of 14 N- and 2 O-glycans was performed by 500 or 600 MHz1H NMR spectroscopy. Besides conventional N-glycans established earlier for human serum transferrin (hST), new (alpha1-3)-fucosylated N-glycans were found, representing sialyl Le(x) elements. Furthermore, as compared to hST, a higher degree of (alpha1-6)-fucosylation and an increase in branching from di- to triantennary compounds has been detected. The presence of O-glycans is demonstrated for the first time in transferrin. PMID- 9751793 TI - The abundance of additional N-acetyllactosamine units in N-linked tetraantennary oligosaccharides of human Tamm-Horsfall glycoprotein is a donor-specific feature. AB - Previously, treatment of Tamm-Horsfall glycoprotein (THp) from different donors with endo-beta-galactosidase has been shown to liberate a tetra- and a Sd(a) active pentasaccharide, concluding the presence of N-linked carbohydrate chains containing additional N -acetyllactosamine units. These type of oligosaccharides were not found in a detailed structure elucidation of the carbohydrate moiety of THp of one male donor, suggesting a donor-specific feature for these type of structures. Therefore, THp was isolated from four healthy male donors and each subjected to endo-beta-galactosidase treatment in order to release these tetra- and Sd(a)-active pentasaccharide. Differences were observed in the total amount of released tetra- and Sda-active pentasaccharide of the used donors (42, 470, 478, 718 microg/100 mg THp), indicating that the presence of repeating N acetyllactosamine units incorporated into the N-glycan moiety of THp is donor specific. Furthermore, a higher expression of the Sd(a) determinant on antennae which display N-acetyllactosamine elongation was observed, suggesting a better accessibility for the beta-N-acetylgalactosaminyltransferase. In order to characterize the N-glycans containing repeating N-acetyllactosamine units, carbohydrate chains were enzymatically released from THp and isolated. The tetraantennary fraction, which accounts for more than 33% of the total carbohydrate moiety of THp, was used to isolate oligosaccharides containing additional N -acetyllactosamine units. Five N-linked tetraantennary oligosaccharides containing a repeating N-acetyllactosamine unit were identified, varying from structures bearing four Sd(a) determinants to structures containing no Sd(a) determinant (see below). One compound was used in order to specify the branch location of the additional N-acetyllactosamine unit, and it appeared that only the Gal-6' and Gal-8' residues were occupied by a repeating N acetyllactosamine unit. PMID- 9751794 TI - Identification of the glycosidically bound sialic acid in mucin glycoproteins that reacts as "free sialic acid" in the Warren assay. AB - A widely employed colorimetric assay for sialic acids based on periodate oxidation followed by reaction with thiobarbituric acid depends on the formation of a hexos-5-uluronic acid product, the pre-chromogen, by the periodate cleavage of the C6-C7, C7-C8, and C8-C9 bonds in free sialic acid. Glycosidically bound sialic acids are not expected to react in the assay since cleavage cannot occur between C6-C7 to yield the pre-chromogen. However, several investigators have reported the detection of a positive reaction by certain sialoglycoconjugates. In this study, it was found that various mucins but not other classes of sialoglycoconjugates or asialomucins exhibited this phenomenon. Of the mucins tested, ovine submaxillary mucin showed the maximum reactivity followed by the bovine and porcine counterparts. The disaccharide Neu5Acalpha2-->6 GalNAc(OH) released from mucins by alkaline borohydride treatment also reacted, albeit weakly compared to the native mucins, but other sialyl saccharides including 6' sialyllactose and 6'-sialyl N -acetyllactosamine did not react. The positive reaction of the submaxillary mucins is not due to the presence of 3-deoxy-d glycero-d-galacto-2-nonulosonic acid (KDN), a minor component in submaxillary mucins, or the release of sialic acid by the acidic condition of the assay. It is demonstrated that sialyl residues linked alpha2-->6 to unsubstituted N acetylgalactosamine (sialyl Tn antigen structure) in mucin glycoproteins is responsible for the positive reaction. Apparently, periodate oxidation of the N acetylgalactosamine residue leads to the release of sialic acid from the Neu5Acalpha2-->6 GalNAc linked to serine/threonine by an acid-catalyzed beta elimination reaction. The findings provide a basis for the development of a chemical method to estimate sialyl Tn epitopes associated with cancer cells. PMID- 9751795 TI - All pyruvylated galactose in Schizosaccharomyces pombe N-glycans is present in the terminal disaccharide, 4, 6-O-[(R)-(1-carboxyethylidine)]-Galbeta1,3Galalpha1 . AB - The large N-linked oligosaccharides released from Schizosaccharomyces pombe by endo-beta-N-acetylglucosaminidase H were examined to determine how the negatively chargedpyruvylated galactoses present (Gemmill,T.R., and Trimble,R.B., 1996, J. Biol. Chem ., 271, 25945-25949) were attached to the oligosaccharide chains. Binding of biotinylated human serum amyloid P and peanut agglutinin to native and depyruvylated S.pombe glycoproteins, respectively, indicated that the pyruvylated epitope was likely to be in the beta configuration. Examination by high-field 1H NMR of whole glycans and a disaccharide fragment released from them on partial acid hydrolysis showed that the pyruvylated galactose species was in fact beta1,3 linked to a second galactose, and this occurred an average of five to six times on nominal Gal57Man64GlcNAc N-glycans. The pyruvate-2,(4,6)Gal-beta1,3Gal epitope is chemically similar to acetaldehyde-Galbeta1,3Gal groups found on the glycoproteins from Paramyxovirus-infected bovine kidney cells (Prehm, P., Scheid,A. and Choppin,P.W. ,1979, J. Biol. Chem ., 254, 9669-9677). The 1:1 stoichiometry between pyruvate and beta-linked galactose in these S.pombe glycans indicates that either pyruvate addition to terminal beta1,3Gal is highly efficient or that pyruvylated Gal is transferred en bloc to alpha1,2-linked Gal residues in theN-linked chains. In contradiction to many galactomannan-producing fungi, which add substantial amounts of Gal in the furanose form to their glycoproteins, all detectable Gal in the large S.pombe galactomannans is in the pyranose form, as found in higher eukaryotes. The current work shows that the S.pombe outer chain structure is a poly-alpha1,6Man backbone 2-O-substituted with either Gal or the pyruvylated galactobiose and contains little alpha1,2-linked or 2-O-substituted Man. This is in contrast to the S. cerevisiae outer chain, which is poly-alpha1,6Man substituted with alpha1,2-linked Man sidechains (Ballou,C.E. ,1990, Methods Enzymol , 185, 440-470). PMID- 9751796 TI - Cold-sensitive E-lysis systems. AB - The release of recombinant bacteria into the environment is undesirable because of possible risks associated with the genetically modified organisms. The aim of this study was to establish a cold-sensitive killing system with a lethal gene, activated when bacteria encounter lower environmental temperatures. To obtain cold-sensitive lysis vectors, the lambdacI857 repressor/pR promoter expression system was combined with either the lacI/lacZpo or the phage 434 cI/pR system that control the expression of the lysis gene E of bacteriophage phiX174. Escherichia coli strains harbouring such suicide vectors are able to grow at 37 degrees C, but cell lysis takes place at temperatures below 30 degrees C. By replacing gene E with a beta-galactosidase reporter gene we also showed that the onset of beta-galactosidase activity corresponds with the onset of lysis at 28 degrees C. Results indicate that these newly combined promoter/repressor systems can also be used to confer cold-sensitive expression to any gene of interest. PMID- 9751797 TI - 5'-flanking sequences required for efficient transcription in vitro of 5S RNA genes, in the related nematodes Caenorhabditis elegans and Caenorhabditis briggsae. AB - In the nematode C. elegans, we had previously observed apparent species specificity in 5S RNA transcription. We have now undertaken a further study of 5S RNA gene transcription in this organism and in the related nematode, C. briggsae; the latter was chosen because it might show evolutionarily conserved, functionally important features. Deletion mutagenesis and transcription in vitro, followed by more precise replacements of short blocks of 5' sequence, show that a short, TATA-like sequence at -25 is essential for efficient transcription in vitro of the 1.0-kb C. elegans 5S DNA repeat, and of both C. briggsae 0.7- and 1.0-kb 5S DNA repeats. Internal sequences within the 5S RNA gene appear to be required and can compete for limiting transcription components, whereas 5' flanking sequences do not. These observations suggest that the process of 5S RNA transcription is similar in these nematodes and other higher eukaryotes. PMID- 9751798 TI - Restricted tissue expression pattern of a novel human rasGAP-related gene and its murine ortholog. AB - The mammalian rasGAPs constitute a group of widely expressed proteins involved in the negative regulation of ras-mediated signaling. In this study we have isolated a novel human gene, RASAL (Ras GTPase-activating-like) and its murine ortholog, MRASAL which are most similar to the GAP1 family of rasGAP proteins, based upon the presence and organization of specific conserved domains. Full-length human and murine mRNA sequences are predicted to encode 804 and 799 amino acid polypeptides, respectively. Sequence analysis of these two proteins revealed the presence of two N-terminal calcium-dependent phospholipid binding C2 domains, a conserved GAP related domain (GRD) and a C-terminal pleckstrin homology (PH) domain. Northern blot and mRNA in situ hybridization analyses indicate that RASAL, in contrast to other mammalian rasGAP proteins, has a limited expression pattern; RASAL is highly expressed in the follicular cells of the thyroid and the adrenal medulla and expressed at lower levels in brain, spinal cord and trachea. Human RASAL has been localized by radiation hybrid mapping to chromosome 12q23 24. PMID- 9751799 TI - Structures of transgene loci in transgenic Arabidopsis plants obtained by particle bombardment: junction regions can bind to nuclear matrices. AB - To clarify the molecular structure of the integration sites of transgenes, we used particle bombardment to examine the DNA sequences of transgene loci. Three transgenic Arabidopsis lines gave a single Southern hybridization band with a selectable gene as the probe. Junction regions flanked by the transgenes were cloned by the inverse polymerase chain reaction method, and the characteristics of the DNA sequences of the 10 junction regions were investigated. All but two of these were AT-rich sequences bearing motifs characteristic of a scaffold/matrix attachment region (S/MAR). Calculations showed that seven of them should have a propensity for curvature. An assay of in-vitro binding to tobacco nuclear matrices showed that all the junction regions bound to nuclear matrices and that the two input DNAs did not bind. The 12 chromosome/transgene (CT) junctions in these three transgene loci were investigated. Cleavage sites for topoisomerase I were found at 10 of the 12, near the junction point. The other two junctions had sites within 6bp of the junction point. The sequence near one terminal of the transgene in the transgene loci was compared with that near the other terminal. Short, direct repeats consisting of 4-6bp were present within 10bp of the junction points in the sequence. We speculate that the transgene introduced by particle bombardment is delivered on AT-rich S/MAR that has a propensity for curvature, and then a nucleotide near the short, direct repeat on the transgene is joined near the cleavage sites on the genome for topoisomerase I. PMID- 9751800 TI - Isolation and characterization of the human prosaposin promoter. AB - Prosaposin is a multifunctional protein that encodes four glycoproteins, named saposins A, B, C and D. They participate in the catabolism of glycosphingolipids in lysosomes. When secreted, intact prosaposin may function as a neuritogenic factor. Human and mouse prosaposin displayed similar temporal and spatial regulation of expression. To gain insight into the transcriptional regulation of this locus, the 5' region was characterized from the human prosaposin gene. The putative human promoter was shown to be TATA-less, i.e. it belonged to the TATA less housekeeping gene family. The transcription initiation sites were localized to -23, -27, -31 and -83bp 5' to ATG, compared to -87 and -94bp in the mouse. In SK-N-SH neuroblastoma cells, positive regulatory elements were detected -343 to 813bp upstream of ATG. A negative regulatory region existed between -813 and 2500bp using SK-N-SH, H441 and NS20Y cells. EMSA and DNA-footprint analysis showed that Sp1 and Sp3 are involved in human prosaposin gene regulation. Compared to the mouse promoter, the human promoter is missing a Sp1 cluster within a 310-bp upstream segment, and has AP-1, Oct-1 and two RORalpha sites that are protected from DNaseI by selected nuclear extracts. PMID- 9751801 TI - Sequence and activity of parathyroid hormone/parathyroid hormone-related protein receptor promoter region in human osteoblast-like cells. AB - The parathyroid hormone (PTH)/PTH-related protein (PTHrP) receptor gene has been characterized in various species. The structure of its promoter and the regulation of its expression in human tissues have, however, not been clearly established yet. We characterized the region upstream of the PTH/PTHrP receptor gene and investigated its promoter activity in the human osteoblast-like SaOS-2 cells. In this region, three untranslated exons were localized, U1 and U2 by using a kidney cDNA, and U3 by homology with the mouse gene. In human osteoblast like cells, a distal promoter (P1) was found to be inactive, as evaluated by luciferase reporter gene assays, in contrast to the situation found in human and mouse kidney tissue. A second promoter (P2), previously described in mouse and human kidney, was shown to be active in human osteoblast-like SaOS-2 cells. We found a hitherto uncharacterized promoter (P3), closely upstream of the ATG start codon. The activities of P2 and P3 were not additive. These results provide important information on the structure of the 5' flanking region of the human PTH/PTHrP gene receptor. PMID- 9751802 TI - A new set of positive/negative selectable markers for mammalian cells. AB - Five new positive and negative selectable markers were created for use in mammalian cells. Their negative selectabilities are based on the Thymidine kinase (Tk) gene of Herpes Simplex virus (HSV) or the Cytidine deaminase (codA) gene of E. coli. The markers can be selected positively by their ability to induce either Hygromycin (Hyg), neomycin (neo), puromycin (PAC) or Blasticidin S (BlaS) resistance. With these markers, two complete sets of markergenes are available that induce independent negative selectable phenotypes. PMID- 9751803 TI - Cloning, characterization and tissue-specific expression of the gene encoding bovine keratocan, a corneal keratan sulfate proteoglycan. AB - Keratocan is one of three major keratan sulfate proteoglycans characteristically expressed in cornea. We reported previously the sequence of bovine Kera cDNA. In this study, the complete bovine Kera gene was cloned and sequenced, and its expression pattern was determined. The Kera gene is composed of three exons and two introns that span 8.830kb of the bovine genome. The first exon contains 287 nucleotides of 5'-UTR sequence. Both of the two large introns of 1322 and 4178bp contain (CA)n repeats. The bovine Kera gene has a TATA box that is located 28bp upstream from tsp. Primer extension and S1 nuclease protection analyses were used to determine the major tsp. RPA indicate that cornea and sclera are the two tissues with the highest expression of Ktcn mRNA. This restricted expression in eye tissues, as well as the unique modification of keratocan with long keratan sulfate chains in cornea, suggests that this molecule may be important in developing and maintaining corneal transparency. PMID- 9751804 TI - Molecular characterization of two genes encoding betaine aldehyde dehydrogenase from amaranth. Expression in leaves under short-term exposure to osmotic stress or abscisic acid. AB - A genomic clone (ahybadh4) and a cDNA (ahybadh17) both encoding betaine aldehyde dehydrogenase (BADH; EC 1.2.1.8) were isolated from the plant Amaranthus hypochondriacus L. The ahybadh4 gene extends 9 kilobases (kb) containing 15 exons with an open reading frame (ORF) of 501 amino acids (aa), a 1.3kb 5' untranslated region (UTR) and a 3' UTR of 0.3kb. The ahybadh17 cDNA encodes a BADH isoform of 500aa which contains 10aa substitutions with respect to AHYBADH4. Both encoded proteins share 98% identity at the amino acid level. Comparison of amaranth BADHs with other reported sequences showed high similarity. Analysis of ahybadh17 expression in amaranth leaves showed that mRNA and BADH protein are present in non-treated amaranth leaves and both transiently increased under short-term exposure to abscisic acid (ABA) and osmotic stress treatments. PMID- 9751805 TI - Cathepsin L gene organization in crustaceans. AB - The gene structure of a cathepsin L of the shrimp Penaeus vannamei has been determined by the polymerase chain reaction. It comprises six exons of various lengths spanning a total of 1792bp. This architecture is homologous to that of rat cathepsin L, three conserved sites of intron position have been effectively identified, with the exception of the third intron break-point located immediately after the cysteinyl active site. In contrast, no similarity is observed with Drosophila or Plasmodium cathepsin L-like gene organizations. This gene expresses a major cathepsin L enzyme in the hepatopancreas. The last intron is polymorphic, suggesting the presence of at least three different genes. PMID- 9751806 TI - The CARE-2 and rel-2 repetitive elements of Candida albicans contain LTR fragments of a new retrotransposon. AB - CARE-2 and Rel-2 are dispersed, repetitive elements of Candida albicans. Hybridisation experiments suggest that they are present at 10-20 copies per genome and appear on most, if not all, of the chromosomes. A high degree of interstrain variation has been demonstrated for CARE-2, making it of use for strain typing. Until now, however, the nature of the repetitive elements within CARE-2 and Rel-2 was unknown. We show here that CARE-2 and Rel-2 contain long terminal repeat (LTR) fragments of a new retrotransposon. These LTRs, which we designate kappa, are partially responsible for the repetitive nature of CARE-2 and Rel-2. Complete copies of the kappa elements are present elsewhere in the genome and adjacent to some are sequences characteristic of the internal regions of retrotransposons. An apparently high degree of scrambling of the kappa elements suggests that they may represent a hotspot for mutation and recombination in C. albicans. PMID- 9751807 TI - Molecular cloning and characterization of a highly conserved chicken cellular nucleic acid binding protein cDNA. AB - A chicken cellular nucleic acid binding protein (cCNBP) cDNA was isolated from a chicken Con-A-stimulated immune cell library by differential screening. cCNBP is a Cys/Cys-His/Cys zinc finger DNA binding protein of unknown function. The chicken CNBP nucleotide and deduced amino acid sequence showed extraordinary sequence conservation (between 81-98% similarities) when compared to human, mouse and rat CNBP. The CNBP gene was shown to be a single copy and to cross-hybridize to human and mouse genomic DNA. A Northern blot analysis revealed cCNBP to be a constitutively expressed gene in a wide variety of tissues and to be differentially expressed in cultured chicken spleen and bursal cells after mitogen stimulation. PMID- 9751808 TI - Genetic analyses of the green visual pigments of rabbit (Oryctolagus cuniculus) and rat (Rattus norvegicus). AB - We have cloned and sequenced the green opsin genes of rabbit and rat. When these genes are expressed in cultured cells and reconstituted with 11-cis retinal, the resulting visual pigments have wavelengths of maximal absorption (lambdamax) of 509nm. These blue-shifted lambdamax values are fully explained by their aa composition A, Y, Y, T, and S at sites 180, 197, 277, 285, and 308, respectively. The inference on the ancestral pigment sequences strongly suggests that the rabbit and rat (and mouse) green pigments attained the extant lambdamax values independently, mainly by the single aa replacement A308S. PMID- 9751817 TI - Evolution and Locomotion. PMID- 9751809 TI - Identification and cloning of a sequence homologue of dopamine beta-hydroxylase. AB - We have identified and cloned a cDNA encoding a new member of the monooxygenase family of enzymes. This novel enzyme, which we call MOX (monooxygenase X; unknown substrate) is a clear sequence homologue of the enzyme dopamine beta-hydroxylase (DBH). MOX maintains many of the structural features of DBH, as evidenced by the retention of most of the disulfide linkages and all of the peptidyl ligands to the active site copper atoms. Unlike DBH, MOX lacks a signal peptide sequence and therefore is unlikely to be a secreted molecule. The steady-state mRNA levels of MOX are highest in the kidney, lung, and adrenal gland, indicating that the tissue distribution of MOX is broader than that of DBH. Antisera raised to a fusion protein of MOX identifies a single band of the expected mobility by Western blot analysis. MOX mRNA levels are elevated in some fibroblast cell strains at replicative senescence, through this regulation is not apparent in all primary cell strains. The gene for MOX resides on the q arm of chromosome 6 and the corresponding mouse homolog has been identified. PMID- 9751818 TI - Gorilla Behaviour. PMID- 9751819 TI - Primate Conservation. PMID- 9751810 TI - Chromosome breakpoints near CpG islands in double minutes. AB - Double minute chromosomes (DMs) are the principal genetic vehicles for amplifying oncogenes in human tumors and drug resistance genes in cultured mouse cells. Mouse EMT-6 cells resistant to methotrexate (MTX) generally contain circular DMs, approximately 1 megabase (Mb) in size, that amplify the dihydrofolate reductase (DHFR) gene. The 1 Mb DMs generally have CpG islands located 500 kb upstream of the DHFR gene. The purpose of this study was to determine the relationship between CpG islands and chromosomal breakpoints giving rise to the DM. We show that EMT-6 cells growing in very low levels of MTX that do not yet contain the 1 Mb DHFR-amplifying DM, develop a NotI/EagI site 500 kb upstream of the DHFR gene. This NotI site is close to, if not identical with, one of the chromosomal breakpoints giving rise to the DM. We show that 500 kb of DM DNA from upstream of the DHFR gene is derived from 500 kb of chromosomal DNA upstream of the chromosomal DHFR gene. The downstream breakpoint maps to a region approximately 200 kb downstream of the DHFR gene near a chromosomal SstII/EagI site. Therefore, approximately 700 kb of DM DNA was derived from the genomic region surrounding the DHFR gene. To confirm the organization of the DM DNA, we isolated DNA probes from the 1 Mb DM. Using pulsed field gel electrophoresis and Southern hybridization, we determined the approximate location of each probe with respect to the CpG island in both the DM and the chromosome. Approximately 300 kb of chimeric DNA from a region unrelated to the DHFR gene was incorporated during DM formation. Implications for the mechanism of DM formation are discussed. PMID- 9751821 TI - Cognition - Lateralization - Motor Behaviour. PMID- 9751820 TI - Communication in Primates. PMID- 9751822 TI - Social Behaviour and Evolutionary Strategies. PMID- 9751823 TI - Evolution and Nutrition. PMID- 9751824 TI - Miscellaneous. PMID- 9751825 TI - Poster Sessions. PMID- 9751827 TI - Pollination of Ravenala madagascariensis and Parkia madagascariensis by Eulemur macaco in Madagascar. AB - Primates are known to be important in dispersal of seeds of tropical rainforest trees, but their role in pollination is very poorly documented. Although the 'traveller's palm' Ravenala madagascariensis is widespread, only a single well documented report exists for its pollination by Malagasy prosimians. Black lemurs, Eulemur macaco, exploit nectaries of Parkia as well as Ravenala systematically at the massifs of Ambato and Lokobe, and almost certainly contribute substantially to their pollination, confirming a proposal first made by Sussman and Raven [Science 1978;200: 731-736]. PMID- 9751826 TI - The evolution of male-infant interactions in the tribe Papionini (Primates: Cercopithecidae). AB - In Old World monkeys, intense affiliative interactions between adult males and infants have mostly been observed in the tribe Papionini. Although these male infant interactions have been reported in most species of the genera Papio, Theropithecus and Cercocebus, they have only erratically been reported in the genus Macaca. In this article I show that the distribution of male-infant interactions within the genus Macaca can be accounted for by the phylogenetic relations among macaque species and by the evolution of the genus Macaca relative to the other Papionini. PMID- 9751828 TI - Group Size and Population Density of the Black Howler Monkey (Alouatta pigra) in Muchukux Forest, Quintana Roo, Mexico. PMID- 9751830 TI - Prey Size Selection under Conditions of Competitive Foraging in Captive Chimpanzees. PMID- 9751829 TI - Growth of marmoset monkeys Callithrix jacchus in captivity. PMID- 9751831 TI - A Preliminary Study on the Variables Correlated with the Emission of Loud Calls in Wild Moor Macaques (Macaca maurus). PMID- 9751832 TI - Interindividual Spatial Proximity in Two Captive Groups of Western Lowland Gorillas (Gorilla gorilla gorilla). PMID- 9751833 TI - Spontaneous Pointing Behaviour in the Wild Pygmy Chimpanzee (Pan paniscus). PMID- 9751834 TI - Stick Throwing by Gorillas (Gorilla gorilla gorilla) at the San Diego Wild Animal Park. PMID- 9751835 TI - Chromosomal variability in the genus Nycticebus. PMID- 9751836 TI - Sexual selection and evolution of the seminal vesicles in primates. PMID- 9751837 TI - A cytogenetic study of Microcebus myoxinus. PMID- 9751838 TI - Observations of Paternal Care in Perodicticus potto at the Cincinnati Zoo and Botanical Garden. PMID- 9751839 TI - Spontaneous Tool Use by a Tonkean Macaque (Macaca tonkeana). PMID- 9751840 TI - Preliminary Field Data on Group Size, Diet and Activity in the Alaotran Gentle Lemur Hapalemur griseus alaotrensis. PMID- 9751841 TI - Additional Data on the Distribution of Cercopithecus (lhoesti) solatus. PMID- 9751842 TI - Determinants and mechanisms of human immune responses to bee venom phospholipase A2. AB - The elicitation of an immune response to protein antigens depends on the specific recognition of antigenic determinants (epitopes) by T and B lymphocytes. Bee venom phospholipase A2 (PLA) represents the major antigen/allergen of honey bee venom. It displays three dominant immunogenic peptide and one glycopeptide T cell recognition sites. These epitopes are equally recognized by both allergic and nonallergic individuals. A mixture of the three epitope containing peptides was successfully used in specific immunotherapy of bee venom-allergic patients. Both peptide and whole bee venom immunotherapy induced a state of specific anergy in T cells. The production of specific IgE and IgG4 antibodies directly correlated with the secreted interleukin-4:gamma-interferon (IL-4:IFNgamma) ratio, which itself depended on the concentration of available antigen and the strength of the T cell-activating signal. This signal comprises accumulated molecular interactions delivered by engagement of the antigenic peptide/MHC class II complex with the T cell receptor (TcR). Indeed the thermodynamic laws of chemical equilibrium reactions reveal that the antigen concentration, together with the equilibration constant Ki and the related Gibbs standard free energy DeltaG degrees of the MHC-II/Ag/TcR complex reaction, may govern the secreted IL 4:IFNgamma ratio, and in consequence, differential IgE and IgG4 antibody formation. Ki includes epitope and MHC-II haplotype variability and therefore represents a measure of immunological individuality. A major B cell epitope was determined by using point-mutated PLA. Specific antigen recognition by B cells can trigger distinct cytokine profiles in T cells and contribute to the differential regulation of specific IgE and IgG4 antibodies. Our results indicate that distinct cytokine profiles inducing allergic and nonallergic responses can be attributed to thresholds of T cell activation generated by the specific binding properties of individual MHC-II molecules to immunogenic T cell epitopes and their presentation to TcR. PMID- 9751843 TI - Simple understanding and optimistic strategy for coping with atopic diseases. IL 5 central hypothesis on eosinophilic inflammation. AB - In this article, accumulated investigations were reviewed to present an 'IL-5 central hypothesis' for eosinophilic inflammation. In this simple working hypothesis, mechanisms of clinical amelioration of atopic diseases manifested by glucocorticoids or immunosuppressants such as cyclosporin A or FK506 were easily understood to stem from the inhibition of IL-5 production and gene transcription. Screening of macrolide molecules resulted in the detection of nonactin (code name: OM-01), which inhibits production and gene transcription of IL-5 but not of IL-2 or IL-4. It was suggested that this kind of approach may open a new era of 'specific gene transcription control' for the management of allergic and other human disorders. PMID- 9751844 TI - Allergy to bovine beta-lactoglobulin: specificity of human IgE using cyanogen bromide-derived peptides. AB - BACKGROUND: Bovine beta-Lactoglobulin (Blg) is a major allergen involved in allergy to cow's milk proteins. Hydrolyzing Blg did not totally suppress its allergenicity; moreover its immunoreactivity may be increased. The aim of this work was to evaluate the specificity of serum IgE to different fragments of Blg in a group of 19 individuals allergic to cow's milk. METHODS: This study was performed using both direct and competitive inhibition ELISA involving immobilized native protein or peptides derived from Blg cyanogen bromide cleavage. RESULTS: Analyses of responses to each peptide revealed a large number of epitopes recognized by specific IgE of human allergic sera. However, there were differences in the specific determinants recognized, depending on the serum. Generally, peptides (25-107) and (108-145) retained substantial proportions of the immunoreactivity of the whole protein. Two other peptides, i.e. (8-24) and (146-162), were less recognized but were not inert. CONCLUSION: The main conclusion is that many epitopes were identified all along the Blg sequence by specific anti-Blg IgE from allergic humans. PMID- 9751845 TI - Cellular and molecular characterization of a major soybean allergen. AB - Soybean proteins share a large number of cross-reacting allergens with other members of the legume family; however, soy-allergic patients rarely react clinically to other members of the legume family. Gly m Bd 30K, an IgE-binding protein with a molecular weight of 30 kD, was identified in soybean extracts by Western IgE-immunoblot analysis. This monomeric allergen was shown to have an N terminal amino acid sequence and amino acid composition identical to that of the seed 34-kD protein, P34, a thiol protease of the papain family. Electron microscopic immunolocalization of P34 monoclonal antibodies and IgE binding to sections of soybean seeds showed dense staining throughout the vacuolar bodies, localizing the allergens in protein storage vacuoles of seed cotyledons. We used pooled serum from soybean-sensitive patients to determine the linear IgE-specific epitopes in the 34-kD allergen amino acid sequence. B-cell epitope mapping revealed 10 regions of IgE-binding activity using an overlapping peptide strategy of 15-mers offset by 8 amino acids throughout the P34 sequence. Smaller overlapping peptides, 10-mers offset by 2 amino acids, revealed 16 distinct linear epitopes, 9 of which were mapped to the mature protein. No obvious amino acid sequence motifs could be identified by the smaller IgE-binding epitopes. Using individual patient serum, 5 immunodominant epitopes were identified in this allergen. PMID- 9751846 TI - Analysis of the cross-reactivity between BtM and Der p 5, two group 5 recombinant allergens from Blomia tropicalis and Dermatophagoides pteronyssinus. AB - BACKGROUND: In tropical climates, sensitization to Bloma tropicalis and Dermatophagoides pteronyssinus is high and mainly directed to species-specific allergens. There is some cross-reactivity between extracts of these mites, probably due to the group 5 allergens that have high sequence homology. OBJECTIVE AND METHODS: We used the radioallergosorbent test (RAST), RAST inhibition and immunoblotting inhibition experiments to investigate the cross-reactivity between the recombinant allergens BtM and Der p 5, expressed as glutathione S-transferase fusion proteins, to detect the epitopes involved and to analyze the importance of this cross-reactivity. RESULTS: Seventy-nine percent of 48 patients sera were RAST positive to both recombinants, with a strong correlation (r = 0.8, p<0.0001). BtM inhibited 25 and 21.1% of IgE-binding to B. tropicalis and D. pteronyssinus extracts respectively and Der p 5 inhibited 22 and 24% of IgE binding to D. pteronyssinus and B. tropicalis extracts. Furthermore, BtM inhibited 74.5% of IgE binding to Der p 5 and Der p 5 inhibited 72.4% of IgE binding to BtM. RAST inhibition with BtM-derived synthetic peptides showed that peptide 4 (residues 35-50) and peptide 5 (residues 46-61) inhibited 37 and 16% of IgE-binding to BtM while peptides 5 and 2 (residues 14-30) were able to inhibit the IgE binding (32 and 28%, respectively) to Der p 5. CONCLUSION: There is cross reactivity between BtM and Der p 5, which explains almost all the cross reactivity between the two mite extracts. This cross-reactivity seems to be related to epitope(s) at the C-terminal segment of these allergens. PMID- 9751847 TI - Evaluation of IgG RAST FEIA for the assay of venom-specific IgG antibodies during venom immunotherapy. AB - BACKGROUND: Successful venom immunotherapy (VIT) in Hymenoptera allergy is usually associated with a strong increase in venom-specific serum IgG antibodies (sIgG). METHODS: We evaluated a new commercial test for the assay of sIgG (Pharmacia CAP Systemtrade mark IgG RAST(R) FEIA; FEIA), in comparison with a conventional ELISA technique. Sera from 40 bee- and 40 Vespula-allergic patients were analyzed by FEIA and ELISA before and 3 months after starting VIT. RESULTS: The correlation between sIgG obtained with the two methods was significant: r = 0.862, p<0.0001 for bee venom (BV), r = 0.861, p<0.0001 for Vespula venom (VV). The geometric mean values obtained with FEIA were higher for both venoms (BV p = 0.03; VV p<0. 01). A highly significant increase (p<0.0001) was observed during VIT with both methods. This increase was concordant in 93% of VV- and 90% of BV treated patients. Intra- and interassay relative coefficients of variation were below 10% for FEIA. CONCLUSION: IgG RAST FEIA is a reproducible and sensitive method for the assay of venom-specific sIgG. PMID- 9751848 TI - Food restriction-mediated adrenal influences on antigen-induced bronchoconstriction and airway eosinophil influx in the guinea pig. AB - The aim of this study was to measure the effects of food restriction on antigen induced bronchoconstriction and inflammatory cell influx in guinea pigs and to determine the role of plasma cortisol and catecholamine concentrations. Ovalbumin (OA; 0.3 mg/kg, i.v.) was administered to OA-sensitized, anesthetized guinea pigs which had been allowed free access to food or had been food restricted for 18 h prior to OA challenge. In addition to higher plasma levels of epinephrine (30% increase) and cortisol (33% increase), fasted guinea pigs had significantly lower (60% decreased) maximal bronchoconstrictor responses to OA than nonfasted, sensitized litter mates. Additionally, groups of fasted or fed animals were subdivided into two additional treatment groups: (1) saline-pretreated or (2) polyethylene glycol 400 (PEG)-pretreated (1 ml/kg, p.o., 1 h prior to antigen challenge). In saline-treated, fasted animals, bronchoconstrictor responses to antigen were significantly diminished (67% decreased) and epinephrine and cortisol levels were increased (64 and 34%, respectively) compared to the corresponding fed group. In both fasted and fed groups, the PEG-treated guinea pigs had higher plasma epinephrine and cortisol levels than animals which received saline, but no significant differences were detected within the PEG treated group. Plasma norepinephrine concentrations were lower in all fasted groups. In a separate model in conscious guinea pigs, there were no differences in aerosol OA-induced bronchoconstriction and eosinophil influx between fasted and fed groups. However, compared to the saline pretreatment group, PEG administration reduced the antigen-induced bronchoconstriction and eosinophilia in both fed and fasted guinea pigs. We speculate that the reduced responsiveness to antigen in fasted versus fed animals may result from food-restriction-induced, stress-related release of epinephrine and cortisol from the adrenal glands, thereby suppressing mast cell degranulation or reducing responsiveness to spasmogenic and chemotactic mediators. In addition, the results suggest that oral dosing with 100% PEG may enhance this phenomenon. PMID- 9751849 TI - RANTES production by cytokine-stimulated nasal fibroblasts: its inhibition by glucocorticoids. AB - Nasal fibroblasts play an important role in both nasal polyposis and nasal allergic diseases and they are known to release a number of proinflammatory cytokines, including GM-CSF, IL-8 and IL-6. The aim of this present work was to investigate whether cytokine-stimulated nasal fibroblasts release biologically active RANTES as well as to study the effect of corticosteroids on the ability of nasal fibroblasts to produce the cytokine. Measurements of RANTES by ELISA demonstrated that RANTES is constitutively secreted spontaneously (21+/-4 vs. 19+/-6 ng/ml, respectively p>0.05). Stimulation of these cells with either TNF alpha, IL-1beta or IFN-gamma induce further release of RANTES in a dose-dependent manner with TNF-alpha being the most potent stimulus. RANTES mRNA expression in nasal fibroblasts correlated with the amount of protein released in the culture supernatant upon cytokine stimulation. Moreover, chemotaxis studies demonstrated that the nasal-derived RANTES was biologically active on eosinophils. Betamethasone and hydrocortisone were found to downregulate RANTES mRNA expression in TNF-alpha-stimulated fibroblasts. These observations suggest that RANTES released by nasal fibroblasts may regulate eosinophil recruitment in nasal disease while glucocorticoids may inhibit the influx of these cells by suppressing the production of RANTES. PMID- 9751850 TI - Expression of adhesion molecules in cultured human nasal mucosal microvascular endothelial cells activated by interleukin-1 beta or tumor necrosis factor-alpha: effects of dexamethasone. AB - Adhesion molecules of microvascular endothelial cells play a key role in the inflammatory processes involved in nonallergic sinusitis. We investigated the cytokine-regulated expression of intercellular adhesion molecule-1 (ICAM-1), E selectin and vascular cell adhesion molecule-1 (VCAM-1), and the effect of dexamethasone on these expressions in cultured human nasal microvascular endothelial cells (HNMEC). ICAM-1 was enhanced, and E-selectin and VCAM-1 were induced in a dose-dependent fashion following stimulation with IL-1beta or TNF alpha. HNMEC differed from human umbilical vein endothelila cells in that (1) maximal upregulation of ICAM-1 expression induced by IL-1beta or TNF-alpha required more time (2) TNF-alpha was more potent than IL-1beta in VCAM-1 expression, and (3) dexamethasone inhibited the upregulation of E-selectin expression alone. These findings contribute to a better understanding of the characteristic features of leukocyte infiltration into inflamed tissue and the effect of glucocorticoid in nonallergic chronic sinusitis. PMID- 9751851 TI - Macrophage inhibition of lymphocyte and tumor cell growth is mediated by 25 hydroxycholesterol in the cell membrane. AB - We have previously reported that a lipid molecule in the membrane fraction of cloned macrophage hybridomas inhibited the growth of lymphocytes and several tumor cell lines. In this study, the inhibitory lipid molecule in the membrane fraction of macrophages was analyzed by thin-layer chromatography and identified as 25-hydroxycholesterol, a family of oxysterols. This conclusion was confirmed by analysis using gas chromatography-mass spectrometry. In addition, both 25 hydroxycholesterol and the lipid molecule recovered from macrophage cell membrane induced apoptosis of the murine T cell lymphoma, BW-5147. These results suggest that an oxysterol expressed in the macrophage cell membrane may participate in the regulation of cell growth through cell contact. PMID- 9751866 TI - Purification and assay of myosin V. PMID- 9751867 TI - Use of latrunculin-A, an actin monomer-binding drug. PMID- 9751868 TI - Use of the F-actin-binding drugs, misakinolide A and swinholide A. PMID- 9751869 TI - F-actin blot overlays. PMID- 9751870 TI - Purification and assay of the Arp2/3 complex from Acanthamoeba castellanii. PMID- 9751871 TI - Purification and assay of the platelet Arp2/3 complex. PMID- 9751872 TI - Purification and assay of zyxin. PMID- 9751873 TI - Purification and assays for paxillin. PMID- 9751874 TI - Assay and purification and focal adhesion kinase. PMID- 9751875 TI - Purification and assays of vasodilator-stimulated phosphoprotein. PMID- 9751876 TI - Listeria monocytogenes-based assays for actin assembly factors. PMID- 9751877 TI - Preparation and characterization of caged fluorescein tubulin. PMID- 9751878 TI - Purification of novel kinesins from embryonic systems. AB - Several kinesin holoenzymes, including the heterotrimeric kinesin-II and bipolar KLP61F complexes described here, are being purified in our laboratory using microtubule affinity precipitation and conventional biochemical fractionation procedures. These protocols have been optimized by using pan-kinesin peptide antibodies and subunit-specific antibodies to monitor the enrichment of kinesin related polypeptides in particular fractions by immunoblotting. Protein purification represents the most direct route available for determining the oligomeric state and subunit composition of a kinesin holoenzyme, for identifying tightly associated accessory subunits such as SpKAP115, and for determining the molecular architecture and functional properties of native kinesin motors. Protein purification methods therefore represent an important complementary approach to molecular genetic approaches that are being pursued in many other laboratories. PMID- 9751879 TI - Assaying processive movement of kinesin by fluorescence microscopy. PMID- 9751880 TI - Purification of dynactin and dynein from brain tissue. PMID- 9751881 TI - Isolation and characterization of kinectin. PMID- 9751882 TI - Purification and assay of CLIP-170. PMID- 9751883 TI - Purification and assay of the microtubule-severing protein katanin. PMID- 9751884 TI - Purification and assay of gamma tubulin ring complex. PMID- 9751885 TI - Rapid isolation of centrosomes. PMID- 9751886 TI - Photoaffinity labeling approach to map the Taxol-binding site on the microtubule. PMID- 9751887 TI - Use of drugs to study role of microtubule assembly dynamics in living cells. PMID- 9751889 TI - Purification, assembly, and localization of FtsZ. PMID- 9751888 TI - Purification and assay of a septin complex from Drosophila embryos. PMID- 9751890 TI - Purification and assembly of FtsZ. PMID- 9751891 TI - High-resolution video and digital-enhanced differential interference contrast light microscopy of cell division in budding yeast. PMID- 9751892 TI - Production of M-phase and I-phase extracts from mammalian cells. PMID- 9751893 TI - Organelle motility and membrane network formation in metaphase and interphase cell-free extracts. PMID- 9751894 TI - Organelle motility and membrane network formation from cultured mammalian cells. PMID- 9751895 TI - In vitro motility assay for melanophore pigment organelles. PMID- 9751896 TI - Purification of phagosomes and assays for microtubule binding. PMID- 9751897 TI - Magnetic bead assay for characterization of microtubule-membrane interactions. PMID- 9751899 TI - Reactivatable cell models of the giant amoeba Reticulomyxa. PMID- 9751898 TI - Reactivation of vesicle transport in lysed teleost melanophores. PMID- 9751900 TI - Design and use of the centrifuge microscope to assay force production. PMID- 9751901 TI - Motility assays on molluscan native thick filaments. PMID- 9751902 TI - Use of optical traps in single-molecule study of nonprocessive biological motors. PMID- 9751903 TI - Versatile optical traps with feedback control. PMID- 9751904 TI - Preparation of a flexible, porous polyacrylamide substrate for mechanical studies of cultured cells. PMID- 9751905 TI - Design and use of substrata to measure traction forces exerted by cultured cells. PMID- 9751906 TI - Purification of cytoskeletal proteins using peptide antibodies. PMID- 9751907 TI - Strategies to assess phosphoprotein phosphatase and protein kinase-mediated regulation of the cytoskeleton. PMID- 9751908 TI - Correlative light and electron microscopy of the cytoskeleton of cultured cells. PMID- 9751909 TI - Clinically significant drug interactions with general anesthetics--incidence, mechanims and management. AB - The frequency of adverse drug interactions increases disproportionately with the increase in the number of drugs given to patients. It was shown that 40% of patients given 16 drugs experienced an adverse drug interaction, compared with 5% of patients given fewer than 6 drugs. The magnitude of the drug interaction problem increases substantially in anesthetised patients because of: (i) the increased use of multiple drugs in the preoperative and intraoperative periods; and (ii) the growing population of geriatric patients who, in addition to having diminished drug metabolising capacity, are often prescribed multiplied medications for concomitant medical illness. Drug interactions with volatile and intravenous anesthetics can be divided into those that are pharmacokinetic and pharmacodynamic in nature. Pharmacokinetic interactions occur when the absorption, distribution, metabolism or excretion of a drug is altered by the coadministration of a second drug. Pharmacodynamic interactions involve a change in the pharmacological effect of a drug as a result of the action of second drug at receptor sites. PMID- 9751910 TI - Morphine-midazolam combination doses for presurgical analgesia in children. AB - We have examined the use of presurgical morphine-midazolam combination in 80 children aged 2-10 y undergoing repair of hypospadias. They were allocated randomly, in a double-blind study, to receive one of four morphine-midazolam combination doses (n = 20 each); (group I: 75 microg/kg each) [corrected] (group II: 75 microg/kg [corrected] morphine, 50 microg/kg [corrected] midazolam); (group III: 50 microg/kg [corrected] morphine, 75 microg/kg [corrected] midazolam); (group IV: 50 microg/kg [corrected] each). Drugs were given after induction of anesthesia and before the start of surgery. Observational scoring system, using crying, movement, agitation, posture and localization of pain as scoring criteria, was used to assess the children during their stay in the recovery room together with their sedative and/or analgesic requirement. Pre surgical morphine-midazolam administration produced stable hemodynamic variables with satisfactory postoperative analgesia suggesting 75 microg/kg [corrected] dose of both morphine and midazolam as upper permissible dose, and 50 microg/kg [corrected] each as lower effective dose. PMID- 9751911 TI - Fat embolization--a war experience. PMID- 9751912 TI - Acute chemical pericarditis following celiac plexus block--a case report. PMID- 9751913 TI - Phylogenetic relationships of platyhelminthes based on 18S ribosomal gene sequences. AB - Nucleotide sequences of 18S ribosomal RNA from 71 species of Platyhelminthes, the flatworms, were analyzed using maximum likelihood, and the resulting phylogenetic trees were compared with previous phylogenetic hypotheses. Analyses including 15 outgroup species belonging to eight other phyla show that Platyhelminthes are monophyletic with the exception of a sequence putatively from Acoela sp., Lecithoepitheliata, Polycladida, Tricladida, Trematoda (Aspidobothrii + Digenea), Monogenea, and Cestoda (Gyrocotylidea + Amphilinidea + Eucestoda) are monophyletic groups. Catenulids form the sister group to the rest of platyhelminths, whereas a complex clade formed by Acoela, Tricladida, "Dalyellioida", and perhaps "Typhloplanoida" is sister to Neodermata. "Typhloplanoida" does not appear to be monophyletic; Fecampiida does not appear to belong within "Dalyellioida," nor Kalyptorhynchia within "Typhloplanoida." Trematoda is the sister group to the rest of Neodermata, and Monogenea is sister group to Cestoda. Within Trematoda, Aspidobothrii is the sister group of Digenea and Heronimidae is the most basal family in Digenea. Our trees support the hypothesis that parasitism evolved at least twice in Platyhelminthes, once in the ancestor to Neodermata and again in the ancestor of Fecampiida, independently to the ancestor of putatively parasitic "Dalyellioida." PMID- 9751914 TI - Molecular phylogeny and evolution of the deep-sea fish genus Sternoptyx. AB - A portion of mitochondrially encoded 12S and 16S ribosomal RNA genes were sequenced from all four valid species of the midwater deep-sea fish genus Sternoptyx (Teleostei: Sternoptychidae) and four sternoptychid outgroup taxa. Secondary structure-based alignment resulted in a character matrix consisting of 865 bp of unambiguously aligned, combined sequences of the two genes, which were subjected to phylogenetic analyses using the maximum parsimony and maximum likelihood methods. The resultant tree topologies from the two methods were congruent and supported by various tree statistics. Although the single most parsimonious tree was not statistically different from the two second parsimonious trees, independent morphological evidence from the anal fin pterygiophore configuration and associated structures strongly supported the former as the preferred hypothesis. Mapping of the contemporary geographic distribution patterns of the four species onto the tree suggested that there was a common ancestor of Sternoptyx with a circumglobal distribution, which had been subdivided into southern and northern ancestral populations along 30 degrees S, possibly through some large-scale oceanographic event. There has been no discernible speciation event in the southern population, though the northern population has subsequently speciated into three contemporary species with largely allopatric/microallopatric distributions. PMID- 9751915 TI - Cryptic species in a "living fossil" lineage: taxonomic and phylogenetic relationships within the genus Lepidurus (Crustacea: Notostraca) in North America. AB - Lineages which exhibit little morphological change over geologic time are evolutionarily and ecologically interesting, but often taxonomically difficult. For some morphologically conservative groups, not only is it almost impossible to identify significant changes in fossil forms over time, but the relationships among extant populations are often poorly understood due to lack of known characters which clearly delimit species. Notostracan crustaceans are a classic example of such "living fossils." The paradoxical characteristics of long-term stasis in gross morphology and hypervariability of many individual morphological characters make notostracans an especially taxonomically challenging group. We used molecular and biochemical techniques to investigate the taxonomic and phylogenetic relationships of four nominal species within the genus Lepidurus in North America, three of which had been alternately abandoned, resurrected, or synonymized under a single, globally distributed morphospecies since their original descriptions in the 1800s. Data from a 330-bp sequence of the mitochondrial 12S rDNA gene and from nine allozyme loci consistently indicate five highly genetically divergent clades among the populations we used to represent the four nominal species. Diagnostic molecular characters, magnitudes of genetic divergence among clades, and the fact that these genetically distinct clades have broadly overlapping geographical ranges strongly suggest that the five clades are reproductively isolated species. One of the nominal species (L. couesii) is not monophyletic, but rather consists of two species which are not sister taxa. The other three nominal species (L. lemmoni, L. packardi, and L. bilobatus) are supported as valid phylogenetic species. The best current hypothesis for phylogenetic relationships among the five species is provided by a simultaneous analysis of both 12S rDNA and allozyme data, which places L. bilobatus and L. "couesii"-1 as sister taxa and L. "couesii"-2 as the most basal of all the Lepidurus species included in this study. These results point to the existence of cryptic species within the current classification scheme for Lepidurus, the need for further taxonomic work within the Notostraca in general, and the role that genetic techniques can play in clarifying the systematics of morphologically conservative groups. PMID- 9751917 TI - Molecular phylogeny and evolutionary divergence of North American biological species of Armillaria. AB - Armillaria is a genus of root infecting basidiomycetes, which includes nine North American biological species. Anonymous nucleotide sequences obtained from four different primer pairs were combined to produce a data set which was analyzed phylogenetically. The data indicated that randomly chosen sequences from the genome were capable of resolving the phylogenetic history of species of Armillaria and provided strong support for intraspecies clustering. NABS III and VII formed a significant monophyletic clade, with III being derived from the more broadly distributed NABS VII. Sequences of isolates of NABS V showed a high degree of variation. This variation may be an indication of recent sympatric speciation, with NABS IX and X diverging from a genetically diverse NABS V. NABS I formed a monophyletic clade despite the variation in geographic distance among the isolates. The position of NABS II as ancestral to NABS I was discussed. However, literature evidence favored divergence of NABS II from NABS I, while this study illustrated genetic similarity of NABS II with NABS VI. NABS VI was the most divergent of the North American species and represented the outgroup. A molecular clock of NABS Armillaria was proposed. PMID- 9751916 TI - Rapid speciation, morphological evolution, and adaptation to extreme environments in South African sand lizards (Meroles) as revealed by mitochondrial gene sequences. AB - Data derived from the morphology of the seven species of South African sand lizards, Meroles (Reptilia, Lacertidae), and their outgroups produce a robust estimate of phylogeny when a maximum parsimony approach is applied. The estimate is fully resolved with little character conflict and internal branches are relatively long. This analysis indicates that Meroles is a true clade that includes the aberrant lacertid long separated as Aporosaura anchietae. The tree is pectinate, its successive external branches representing species with increasing adaptation to desert conditions, especially aeolian sand habitats. This pattern, and the robustness of the tree, support a model of invasion of severe habitats in which successive rounds of speciation, displacement, and adaptation result in spread into extreme ecological situations. To test the robust morphological phylogeny and, indirectly, the model as well, DNA from mitochondrial 12S and 16S ribosomal genes was sequenced and analyzed by both maximum parsimony and maximum likelihood approaches. Trees produced were largely congruent with that derived from morphology, although different from ones resulting from protein electrophoresis. However, in contrast to the internal branches of the morphological tree, those of the DNA maximum likelihood tree are quite short. The DNA data provide some corroboration for the relationships within Meroles based on morphology and consequently for the model as well. The disparity in internal branch lengths between the maximum parsimony morphological and maximum likelihood DNA trees may well indicate that the multiple adaptations to desert conditions arising on the main lineage of Meroles evolved quite rapidly. In this study DNA thus not only corroborates the phylogeny but also provides evidence about another aspect of evolutionary history. PMID- 9751918 TI - Phylogenetic relationships of terrestrial Australo-Papuan elapid snakes (subfamily Hydrophiinae) based on cytochrome b and 16S rRNA sequences. AB - Phylogenetic relationships among the venomous Australo-Papuan elapid snake radiation remain poorly resolved, despite the application of diverse data sets. To examine phylogenetic relationships among this enigmatic group, portions of the cytochrome b and 16S rRNA mitochondrial DNA genes were sequenced from 19 of the 20 terrestrial Australian genera and 6 of the 7 terrestrial Melanesian genera, plus a sea krait (Laticauda) and a true sea snake (Hydrelaps). These data clarify several significant issues in elapid phylogeny. First, Melanesian elapids form sister groups to Australian species, indicating that the ancestors of the Australian radiation came via Asia, rather than representing a relict Gondwanan radiation. Second, the two major groups of sea snakes (sea kraits and true sea snakes) represent independent invasions of the marine environment. Third, the radiation of viviparous Australian elapids is much older than has been suggested from immunological data. Parsimony analyses were unable to resolve relationships among the Australian radiation, a problem previously encountered with analyses of other (morphological, electrophoretic, karyotypic, immunological) data sets on these species. These data suggest that the reason for this continued difficulty lies in the timing of speciation events: the elapids apparently underwent a spectacular adaptive radiation soon after reaching Australia, such that divergences are ancient even within genera. Indeed, intrageneric divergences are almost as large as intergeneric divergences. Although this timing means that our sequence data cannot fully resolve phylogenetic relationships among the Australian elapids, the data suggest a close relationship of the following clades: Pseudonaja with Oxyuranus; Ogmodon with Toxicocalamus; Demansia with Aspidomorphus; Echiopsis with Denisonia; the "Notechis" lineage with Drysdalia coronoides; and Rhinoplocephalus and Suta with Drysdalia coronata. At least two of the Australian genera (Drysdalia and Simoselaps) appear to be paraphyletic. These sequence data support many of the conclusions reached by earlier studies using other types of data, but additional information will be needed before the phylogeny of the Australian elapids can be fully resolved. PMID- 9751920 TI - Phylogenetic relationships between oviparous and viviparous populations of an Australian lizard (Lerista bougainvillii, scincidae). AB - Viviparity has evolved from oviparity in many vertebrate lineages, and species that contain both oviparous and viviparous populations offer the best opportunity for a detailed examination of the processes involved in this major life-history transition. However, although several such species have been reported, none have been the subject of detailed phylogenetic analyses. We examine such a case within the Australian scincid lizard bougainvillii. Data were obtained by sequencing a 314-bp segment of mitochondrial cytochrome b from 32 individuals from 17 populations of L. bougainvillii and two morphologically similar congeneric species (L. dorsalis L. microtus). Sequences were aligned and analyzed using parsimony and distance methods. The resultant matriarchal phylogeny resolved the populations of L. bougainvillii into three major groups: a population from NSW; a group predominantly from Eyre Peninsula; and a less well-defined group from the central part of the species range. The NSW and Eyre Peninsula groups are oviparous and are quite divergent from other L. bougainvillii populations and from each other. The central group contains both viviparous and oviparous populations, and seem to represent a more recent radiation within the species. Our results indicate that viviparity has evolved at least twice within the genus Lerista, because the viviparous L. microtus is not closely related to viviparous populations of L. bougainvillii. The lack of phylogenetic separation of mtDNAs from viviparous and oviparous populations within L. bougainvillii relative to strong geographic structure within the latter indicates that populations with different reproductive modes are indeed conspecific. Lerista bougainvillii is thus the first vertebrate species for which intraspecific bimodality in reproductive mode can be claimed with any certainty. PMID- 9751919 TI - Comparing molecular evolution in two mitochondrial protein coding genes (cytochrome b and ND2) in the dabbling ducks (Tribe: Anatini). AB - Rates of sequence evolution were estimated for the cytochrome b (cyt b) and NADH dehydrogenase sub-unit 2 (ND2) genes using a phylogeny of the dabbling ducks (Tribe: Anatini) and outgroups. This speciose group was densely sampled, reducing the impact of undetected homoplasy on rate comparisons. Phylogenies based on sequences of the two gene regions and various weighting schemes differed, but most of the differences involved weakly supported nodes. In addition, partition homogeneity tests show that these differences were not due to statistically significant conflict between the data sets. Cyt b and ND2 also showed similar rates and types of both nucleotide and amino acid substitutions. For both genes, substitutions between isoleucine and valine and between alanine and threonine were most common; both of these substitution types are the result of A-G transitions at first positions of codons. Rates of sequence evolution varied substantially and significantly among nucleotide positions, and even within a given codon position (first, second, or third), rates were significantly heterogeneous among sites. Within Anatini, cyt b and ND2 show similar levels of variation and homoplasy, and are equally useful for reconstructing the species level phylogeny of this group. PMID- 9751921 TI - A molecular perspective on the systematics and evolution of the genus Arvicanthis (Rodentia, Muridae): inferences from complete cytochrome b gene sequences. AB - Systematics of the genus Arvicanthis, the African unstriped grass rat, are somewhat controversial. Most recent taxonomic revisions list five to six species but the definition of some of these (Arvicanthis dembeensis, Arvicanthis nairobae, and Arvicanthis niloticus) is uncertain. The complete mitochondrial cytochrome b gene (1140 bp) was sequenced for 20 specimens from throughout the range of the genus to determine the intrageneric genetic structure, construct a molecular phylogeny, and evaluate classical taxonomies. Neighbor-joining and maximum parsimony analyses yielded identical phylogenetic trees that identify two major lineages: the first one (1) is composed of specimens usually referred to A. niloticus but representing several distinct species, and the other (2) is a complex including "true" A. niloticus from Egypt and northern West Africa as well as Arvicanthis abyssinicus, Arvicanthis dembeensis, and Arvicanthis somalicus. An analysis on a 357-bp fragment of the cytochrome b including published data on A. nairobae indicates that this taxon is part of clade (1). Calibration of the number of 3rd position transversion changes with the murid fossil record suggests that clades (1) and (2) diverged approximately 5 Myr ago. Arvicanthis niloticus as currently recognized is a paraphyletic association and this name should be restricted to the Egyptian and northern West African sample. We also suggest referring to A. dembeensis as A. niloticus, as our cytochrome b data do not support its recognition as a distinct species. Clade (1) is subdivided in three lineages, geographically corresponding to southern West, Central, and East Africa. The high genetic divergence detected between the Central African lineage and the other two lineages suggests that they probably represent separate species. Clade (2) experienced rapid cladogenetic events during the late Pliocene, with the A. somalicus lineage being the first to emerge, followed by the ancestor of A. abyssinicus and A. blicki. This period was characterized by significant climatic and environmental changes, such as the extension of open habitats, which might have provided a stimulus for speciation in this savanna dwelling genus. Confrontation of our molecular results with chromosomal data shows a high degree of congruence between the two datasets. PMID- 9751923 TI - Mitochondrial phylogeny of the European cyprinids: implications for their systematics, reticulate evolution, and colonization time. AB - Two different mitochondrial genes, the cytochrome b and the 16S rDNA, support the same European cyprinid molecular phylogeny: the most basal subfamily is the paraphyletic Rasborinae, the Cyprininae are monophyletic, the Tincinae and Gobioninae are close to the Cyprininae or more basal lineages but not close to Leuciscinae or Alburninae, and the Leuciscinae are paraphyletic but can become monophyletic if we include the biphyletic alburninae and exclude the Phoxinini. The relationship of the Acheilognathinae remains obscure. Natural intergeneric and interspecific hybridizations are clearly demonstrated within the Leuciscinae, both from high bootstrap proportions and intermediate morphological features: Chondrostoma toxostoma and Rutilus rutilus, Scardinius erythrophthalmus and R. rutilus, and Leuciscus multicellus and Leuciscus soufia. Finally, the use of the nonsaturated and clockwise 16S mtDNA sequences have been used to infer from nonintrogressive taxa the time of the first European cyprinid cladogeneses. The estimation confirms the hypothesis of Almaca and Banarescu that European cyprinid subfamilies started to diversify 35 mya and confirms the hypothesis of Bianco on the diversification of European leuciscines in the Mediterranean area during the late Messinian (6.5 to 5.3 mya). PMID- 9751924 TI - Mitochondrial phylogeny of the genus Regulus and implications on the evolution of breeding behavior in sylvioid songbirds. AB - We tested four hypotheses about the relationships of the kinglets (genus Regulus)to seven closely related genera of the songbird superfamily Sylvioidea using mitochondrial DNA sequences. The kinglets were suggested to be closely related to the tits (Parus) or to the Old World Warblers (Phylloscopus) and were also suggested to constitute the, or at least one of the, most ancestral splits among the sylvioids. Our phylogenetic analysis grouped the kinglets as the sister group of a clade comprising Parus and Phylloscopus and including the genera Sylvia, Aegithalos, and Leptopoecile. Two of the taxa were placed more ancestral to the kinglets: Sitta and Certhia. We also identified the endemic kinglet species from the Canary Islands s the sister group of R. regulus. The superimposition of breeding behavior on the phylogeny suggests that hole nesting is ancestral and various other patterns of nest construction have evolved from it. The placement of Parus implies that hole nesting in the Paridae is likely to have originated secondarily. Further, Leptopoecile and Aegithalos, two genera for which a helper system of elder offspring in breeding was described, were resolved as a clade. PMID- 9751925 TI - Bone density measurements and therapeutic decision making. PMID- 9751922 TI - Phylogenetic relationships among Agamid lizards of the Laudakia caucasia species group: testing hypotheses of biogeographic fragmentation and an area cladogram for the Iranian Plateau. AB - Phylogenetic relationships within the Laudakia caucasia species group on the Iranian Plateau were investigated using 1708 aligned bases of mitochondrial DNA sequence from the genes encoding ND1 (subunit one of NADH dehydrogenase), tRNAGln, tRNAIle, tRNAMet, ND2, tRNATrp, tRNAAla, tRNAAsn, tRNACys, tRNATyr, and COI (subunit I of cytochrome c oxidase). The aligned sequences contain 207 phylogenetically informative characters. Three hypotheses for historical fragmentation of Laudakia populations on the Iranian Plateau were tested. In two hypotheses, fragmentation of populations is suggested to have proceeded along continuous mountain belts that surround the Iranian Plateau. In another hypothesis, fragmentation is suggested to have resulted from a north-south split caused by uplifting of the Zagros Mountains in the late Miocene or early Pliocene [5-10 MYBP (million years before present)]. The shortest tree suggest the later hypothesis, and statistical tests reject the other two hypothesis. The phylogenetic tree is exceptional in that every branch is well supported. Geologic history provides dates for most branches of the tree. A plot of DNA substitutions against dates from geologic history refines the date for the north-south split across the Iranian Plateau to 9 MYBP (late Miocene). The rate of evolution for this segment of mtDNA is 0.65% (0.61-0.70%) change per lineage per million years. A hypothesis of area relationships for the biota of the Iranian Plateau is generated from the phylogenetic tree. PMID- 9751926 TI - Volumetric measurements of bone mineral density of the lumbar spine: comparison of three geometrical approximations using dual-energy X-ray absorptiometry (DXA) AB - Measurements of bone mineral density using dual-energy X-ray absorptiometry (DXA) gives area values (g cm-2) rather than true volumetric values (g cm-3). To calculate the vertebral volume using planar postero-anterior and lateral DXA values, several different geometrical approximations were used: cubic, cylindrical with a circular cross-section and cylindrical with an elliptical cross-section. The aim of this study was to compare these geometrical approximations with each other and with a reference standard, defined as the volume found on a computed tomographic (CT) scan. L2 and L3 were evaluated in a phantom study. Volume approximations by the cube or cylinder with circular cross section geometry showed more than a 50% overestimation (range 54-74%). However, the elliptical cylinder approach showed very good agreement: 2.1% and 1.2% for L2 and L3, respectively, when compared to the CT volumes. In addition, we performed four patient studies with both CT and DXA to evaluate the elliptical cylinder estimate in a clinical setting. For L2 and L3, the mean relative difference was less than 2%. We conclude that the elliptical cylinder approach results in the most accurate bone volume estimates in both the phantom and patients. PMID- 9751927 TI - 111In-pentetreotide scintigraphy is superior to 123I-MIBG scintigraphy in the diagnosis and location of chemodectoma. AB - Chemodectomas, or glomus tumours, are unusual head and neck paragangliomas. A non invasive imaging technique, 123I-metaiodobenzylguanidine (123I-MIBG) scintigraphy, has long been used for the diagnosis of all types of paraganglioma. The aim of this study was to evaluate and compare classic 123I-MIBG scintigraphy with the more recent 111In-pentetreotide scintigraphy in the diagnosis and location of chemodectomas. We performed 123I-MIBG and 111In-pentetreotide scintigraphy in eight patients (7 females, 1 male) with histologically or radiologically confirmed chemodectomas (five carotid body and three jugulotympanic chemodectomas). 123I-MIBG uptake was visualized in four patients on planar views and SPET images (sensitivity 50%); uptake was low in three patients. Using 111In-pentetreotide scintigraphy, all chemodectomas in eight patients were visualized (sensitivity 100%) and 111In-pentetreotide uptake was high in all cases. In conclusion, our results indicate that 111In-pentetreotide scintigraphy is superior to 123I-MIBG scintigraphy in the diagnosis and location of chemodectomas. In-pentetreotide or 123I-MIBG uptake suggests a neuroendocrine origin, providing important functional information in the diagnosis of chemodectomas. Moreover, 111In-pentetreotide scintigraphy permits a good classification of patients with or without somatostatin receptors in the chemodectoma in the application of pharmacological therapy with somatostatin analogues to inoperable tumours. The main therapeutic action of cold somatostatin analogues is to inhibit hormonal hypersecretion in different neuroendocrine tumours. In chemodectomas, however, the most important effect of somatostatin analogues is to reduce tumour volume or inhibit growth progression. PMID- 9751928 TI - Dynamic distribution and dosimetric evaluation of human non-specific immunoglobulin G labelled with 111In or 99Tcm. AB - This study presents data on the dynamic distribution and dosimetry of 111In- and 99Tcm-labelled human non-specific immunoglobulin G (IgG), two recently developed radiopharmaceuticals for the detection of infection and inflammation. Five healthy volunteers were injected with 20-75 MBq 111In-IgG and seven patients were injected with 740 MBq 99Tcm-hydrazinonicotinamide derivative (HYNIC)-IgG. Blood samples, urine and feces were collected. Whole-body gamma camera imaging studies were performed. The activity in source organs was quantified using the conjugate view counting method and a partial background subtraction technique. Dosimetric calculations were performed using the MIRD technique. For 111In-IgG, the mean biological half-times in the blood were 0.90 and 46 h for the a- and b-phase, respectively. For 99Tcm-HYNIC-IgG, these half times were 0.46 and 45 h. For 111In IgG, the mean cumulative urinary excretion in the first 48 h was 18% of the injected dose, while excretion in the feces was less than 2% of the injected dose. For 99Tcm-HYNIC-IgG, the whole-body retention was always 100% up to 24 h. The mean absorbed doses in the liver, spleen, kidneys, red marrow and testes from 111In-IgG were 0.8, 0.7, 1.2, 0.3 and 0.4 mGy MBq-1 respectively. The mean absorbed doses for 99Tcm-HYNIC-IgG to these organs were 16, 24, 15, 10 and 22 mu Gy MBq-1 respectively. The mean effective dose was 0.25 mSv MBq-1 and 8.4 mu Sv MBq-1 for 111In-IgG and 99Tcm-HYNIC-IgG respectively. In conclusion, the radiation absorbed doses for both 111In-IgG and 99Tcm-HYNIC-IgG are low and, therefore, these radiopharmaceuticals can be administered safely from a radiation risk perspective. PMID- 9751929 TI - Characterization of late abdominal accumulation of 99Tcm-HMPAO leukocytes in a large population of children. AB - We retrospectively evaluated the incidence of late accumulation of 99Tcm-HMPAO leukocytes (99Tcm-WBC) in the right lower quadrant of a large population of children and characterized some predictive patterns that would enable differentiation of active inflammation from this late occasional accumulation of 99Tcm-WBC. We reviewed the charts of 211 children. The first group evaluated consisted of 79 controls: 30 normal children with no gastrointestinal disease, but who underwent 99Tcm-WBC scanning for other medical problems, and 49 children who had non-specific gastrointestinal (GI) complaints, but had no demonstrable inflammatory bowel disease by conventional diagnostic methods. The second group consisted of 132 children with inflammatory bowel disease: 80 children with Crohn's disease (CD), 34 with ulcerative colitis (UC) and 18 with indeterminate colitis (IC). Children were imaged at 30 min and 3 h. Fifteen (19%) of the 79 controls scanned showed accumulation of 99Tcm-WBC in the right lower quadrant at 3 h and none at 30 min. Of those 15, 8 were from the control population and 7 from the group with non-specific GI complaints and negative work-ups. There was no uptake in other segments of the bowel. The accumulation was faint, of lesser intensity than in the iliac wing, and diffuse, such that identification of a specific loop of involved bowel was not possible. Migration of the 99Tcm-WBC distal to the terminal ileum was demonstrated. The other 64 children in the control group showed no accumulation of 99Tcm-WBC at any time during their scans. All 79 scans were blindly interpreted as normal studies. There were no false positive readings encountered in the 132 children with inflammatory bowel disease (80 CD, 34 UC, 18 IC) when the aforementioned characteristics of the late accumulation of 99Tcm were used to differentiate inflammation from this physiological excretion. In conclusion, the late accumulation of 99Tcm-WBC in the right lower quadrant is characterized by (1) accumulation at no less that 3 h, (2) no accumulation in other segments of the bowel, (3) faint accumulation of lesser intensity than in the iliac wing, (4) a diffuse accumulation pattern and (5) migration of the 99Tcm-WBC into the caecum and ascending colon over time. Recognition of this excretion pattern enables differentiation of active Crohn's disease of the small bowel from migration and accumulation of 99Tcm-WBC in the right lower quadrant of the abdomen. PMID- 9751930 TI - The use of dual-isotope imaging to compare the gastrointestinal transit of food and pancreatic enzyme pellets in cystic fibrosis patients. AB - Cystic fibrosis patients require pancreatic enzyme supplements to aid food digestion. It is suspected that incorrect delivery of this enzyme may result in both significant malabsorption and the development of strictures in the proximal colon caused by the high-dose supplement reaching this region before the food. Investigations into the drug's delivery were performed using dual-isotope imaging; a method was developed to directly label the enteric-coated enzyme pellets with 111In, re-applying the enteric coating afterwards, and this was then ingested with a pancake meal labelled with 99Tcm-tin colloid. Consecutive image data, acquired over a period of > or = 4 h using a dual-headed gamma camera, were analysed to assess intestinal transit. In-vitro stability checks on these labelling techniques were encouraging, showing < 2% 99Tcm and < 7% 111In elution over 90 min in hydrochloric acid. In 5 of the 12 patients studied to date, the pellets were seen to pass through significantly faster than the food, with a mean difference in 50% gastric emptying time of greater than 93 min. The mean absolute difference in emptying time for all 12 patients was > 67 min. Thus, a technique has been developed to effectively radiolabel pancreatic enzyme pellets, and analysis of dual-isotope images using this preparation, together with radiolabelled solid food, has demonstrated significant differences in the transit of these two substances through the gastrointestinal tract of some cystic fibrosis patients. PMID- 9751931 TI - Accuracy and clinical utility of the mini-dose 14C-urea breath test in the evaluation of Helicobacter pylori infection. AB - The aim of this study was to evaluate the accuracy of the 14C-urea breath test by comparing the results to those obtained by endoscopy with mucosal biopsy. We also examined the value of the breath test result obtained prior to endoscopy in predicting peptic ulcer disease. Forty-two individuals underwent the 14C-urea breath test. Collections of expired C02 were analysed using a liquid scintillation counter. All individuals then underwent endoscopy with biopsy. Biopsy material was evaluated by the rapid urease method and by histology for the presence of H. pylori. Our results demonstrated that the 14C-urea breath test was 100% sensitive and specific when compared to the rapid urease test as the 'gold standard' for the detection of H. pylori. In comparison to pathology, the sensitivity remained 100% and the specificity was 89%. The results of the 14C urea breath test had a poor predictive value for the determination of peptic ulcer disease. We conclude that the 14C-urea breath test can be easily performed at any medical facility equipped with a liquid scintillation counter and can accurately detect H. pylori. A negative breath test result could not exclude the presence of peptic ulcer disease. PMID- 9751932 TI - Cerebral hypoperfusion detected by SPET in early neuro-Behcet syndrome. AB - We used brain single-photon emission tomography (SPET) to detect hypoperfused areas in 15 consecutive Behcet syndrome patients. Five were suspected of having neuro-Behcet syndrome, having at least one neurological symptom. For these patients, SPET was performed within 1 month of the onset of nervous system involvement. The 15 patients fulfilled the criteria of the International Study Group for Behcet syndrome. Neurological assessment and SPET were complemented by EEG in all five patients with suspected neuro-Behcet syndrome and by magnetic resonance imaging in three. Brain SPET detected hypoperfused regions in all five neurological patients; EEG showed abnormalities in three. Magnetic resonance imaging was normal in the three patients in whom it was performed. SPET was negative in all patients without neurological involvement and 20 healthy controls. SPET detected a reduction in brain blood flow in early neuro-Behcet syndrome, but there was no definitive correlation between the hypoperfused brain regions and the clinical features. Further studies are required to evaluate the significance of brain hypoperfusion and the value of SPET in the early diagnosis of neuro-Behcet syndrome. PMID- 9751933 TI - Left-right asymmetry of striatal dopamine D2 receptors. AB - Brain functions may be lateralized to the right or the left hemisphere. However, the biochemical characteristics accompanying these functions are largely unknown. To test possible lateralization of striatal dopamine D2 receptors, we examined 18 volunteers using 123I-iodobenzamide and single photon emission tomography. The striatum-to-cerebellum D2 binding ratio was 1.93 +/- 0.22 (mean +/- S.D.) on the right side and 1.85 +/- 0.19 on the left side. In 14 subjects, D2 binding was higher in the right compared to the left striatum (P < 0.05). These results are supported by a meta-analysis performed on 15 studies reported in the literature. We conclude that side differences of striatal dopamine D2 receptors exist. We propose that motor activity could be responsible for our findings. PMID- 9751934 TI - The safety of dipyridamole-thallium imaging in patients with critical aortic valve stenosis and angina. AB - Angina pectoris occurs in many patients with critical aortic valve stenosis, but in less than 50% of cases is it due to atherosclerotic coronary disease. Pre operative coronary angiography is used to determine whether coronary revascularization is required in addition to aortic valve replacement. The aim of this study was to determine the safety and image quality of dipyridamole-thallium imaging (DPT) in excluding coronary artery disease requiring a coronary artery bypass graft in patients with aortic valve stenosis requiring aortic valve replacement. Dipyridamole-thallium imaging and coronary angiography were performed less than one month apart in patients with clinical and echo-Doppler evidence of aortic valve stenosis requiring aortic valve replacement. Coronary angiography and DPT were each interpreted by experienced observers blind to the other result. The safety of DPT was judged by symptoms, ECG changes, haemodynamic effects and the need for stress reversal. Image quality was determined from the myocardial-to-background thallium uptake ratio in a normal segment. Twelve patients with aortic valve stenosis (gradient 95 +/- 24 mmHg) were studied, all of whom had left ventricular hypertrophy. The dominant symptom was angina pectoris in eight patients, syncope in three and dyspnoea in one. None had previous myocardial infarction, but two were smokers, six were hyperlipidaemic and one was hypertensive. The patients tolerated DPT well and only one required stress reversal. The quality of the DPT images was good. The DPT image was entirely normal in eight (66%) patients, none of whom had coronary artery disease. Reversible defects were seen in four patients, all of whom had significant coronary artery disease. We conclude that DPT is safe in patients with aortic valve stenosis and angina pectoris. The image quality is good despite left ventricular hypertrophy. In patients with angina pectoris and aortic valve stenosis, coronary angiography can safely be restricted to those with abnormal myocardial perfusion results. PMID- 9751935 TI - Pharmacokinetics of 99Tcm-pertechnetate and 188Re-perrhenate after oral administration of perchlorate: option for subsequent care after the use of liquid 188Re in a balloon catheter. AB - Radioactive wires and other linear sources are currently being used in clinical trials as endovascular brachytherapy to prevent restenosis after percutaneous transluminal coronary angioplasty. A new concept is the use of a liquid-filled balloon containing a beta-emitting radioisotope. A major advantage is optimal delivery of the radioactivity to the vessel wall. Rhenium-188 (188Re) is a high energy beta-emitter that is routinely available from a 188W/188Re generator in liquid form. Since 188Re-perrhenate could be released in the unlikely event of balloon rupture, we investigated whether, in analogy to pertechnetate, subsequent use of perchlorate can reduce the uptake of perrhenate in the thyroid. We performed static (n = 9) and dynamic (n = 11) thyroid scintigraphy with 99Tcm pertechnetate to estimate the overall reduction in activity within 30 min and the washout from the thyroid after oral administration of 600 mg perchlorate (T1/2). In two patients, 188Re was injected to estimate the whole-body distribution and the discharge of thyroid activity after perchlorate use. Based on MIRD Dose Estimate Report No. 8 (valid for 99Tcm-pertechnetate), the radiation burden was calculated for intravenous administration of 188Re and competitive blocking with perchlorate. In 20 patients, 99Tcm uptake by the thyroid was reduced by 85% within 30 min by perchlorate. The mean (+/- S.D.) washout rate (T1/2) was 8 +/- 2 min in 11 patients. Perrhenate showed a whole-body distribution similar to that of pertechnetate and the thyroid activity could be displaced (T1/2 = 6.3 and 9.3 min, respectively) by oral administration of perchlorate, with reductions in uptake of 83% and 75% within 30 min, respectively. Whole-body scanning demonstrated no regional accumulation of 188Re-perrhenate with excretion by urine. Dose estimates gave an effective dose equivalent of 0.42 mSv MBq-1, which decreased to 0.16 mSv MBq-1 after perchlorate blocking. 188Re has favourable properties for endovascular brachytherapy via a balloon catheter and, in the unlikely event of balloon rupture, whole-body radiation can be reduced to 38% by subsequent oral administration of perchlorate. PMID- 9751936 TI - Hybridization of a 99Tcm-labelled oligodeoxynucleotide to CAPL RNA. AB - Oligodeoxynucleotides (ODNs) labelled with an appropriate radionuclide could provide a means to identify serious diseases early on and thereby help initiate treatment at a very early phase. Regardless of important issues like in-vivo stability and membrane passage, the key issue for the oligonucleotide approach is the ability of the radiolabelled ODN to hybridize to the target mRNA. The secondary structure of mRNA does not permit all complementary ODNs to hybridize and a careful selection of the probe with consecutive testing is therefore necessary. This study was initiated to demonstrate hybridization of a 99Tcm labelled 20-mer ODN to RNA of CAPL (S100A4), a gene reported to be overexpressed in metastatic cancers like breast carcinoma and osteosarcoma. The phosphodiester ODN GX-1 (antisense) and two control sequences (scrambled and random) were conjugated to the bifunctional chelating agent S-benzoyl-mercaptoacetyltriglycine (S-benzoyl-MAG3) and labelled with 99Tcm. The radiolabelled ODNs were purified on a C18 mini-column and characterized on a reverse-phase HPLC system. The radio chemical purity was > 90% and the product was stable for > 6 h in aqueous medium. The hydrization properties of unlabelled, 32P-labelled and 99Tcm-labelled ODNs to transcribed RNA were studied using polyacrylamide gel electrophoresis (PAGE). Direct hybridization of GX-1 to transcribed RNA was demonstrated. A 50-fold excess of unlabelled ODN over transcribed RNA caused a near to complete consumption of RNA by RNase H activation. In 1:1 proportions of radiolabelled (32P and 99Tcm) ODNs to RNA, only radiolabelled GX-1 was found to hybridize to RNA in a PAGE system. The radiolabelled control ODNs did not show signs of hybridization. This study demonstrates that 3'-99Tcm-labelling of ODNs does not interfere with the hybridization properties of the ODNs in solution, making 99Tcm labelling an attractive procedure for the future development of antisense technology in imaging. PMID- 9751938 TI - Use of nitroglycerin for uterine relaxation. AB - Data from human and experimental animal research indicate that nitric oxide (NO), a novel messenger, formed during the nitric oxide synthase-catalyzed oxidation of L-arginine to L-citrulline, is involved in maintaining normal uterine tone during gestation. There are demonstrated and potential benefits of manipulating the L arginine-NO system during pregnancy. Several recent case reports and case series have described the effective use of nitroglycerin (GTN), a NO donor compound, antenatally, intrapartum, and postpartum for acute uterine relaxation. Therapeutic indications for GTN range from facilitating external cephalic version, difficult vaginal or cesarean section delivery, and manual exploration of the uterus, to its use as a tocolytic. The intravenous regimen of GTN required to obtain the desired degree of uterine relaxation is extremely variable; intravenous bolus doses of 50 micrograms to 500 micrograms GTN with up to three repeated injections of 50 micrograms to 250 micrograms have been reported. Other methods of GTN administration include transdermal patches and sublingual spray. GTN, when used in low doses, may provide safe and effective uterine relaxation with no clinically apparent fetal or maternal adverse effects. However, clinical trials with use of objective methods of evaluating uterine tone and comparing GTN to other tocolytic agents are required before widespread use in advocated. PMID- 9751939 TI - Laparoscopic cholecystectomy during pregnancy: a case series and review of the literature. AB - This study was conducted to evaluate the role of antepartum laparoscopic cholecystectomy (LC). Patients who underwent LC were identified from a hospital database with the use of CPT/ICD codes. Of 2093 cases performed at a major center (October 1991 to November 1997), only six were performed during pregnancy. On reviewing the English literature, gestational age at surgery and delivery and outcome of delivery were provided in only 69 of 105 patients (33 papers with 1-10 cases) and we tabulated different variables from the cases in this review. In this series, two patients who had LC in the first trimester underwent elective termination of pregnancy. Of the seven published cases of first trimester LC followed to delivery, one had preterm delivery. First trimester open cholecystectomy (OC) has a 12 percent spontaneous abortion rate. The four patients who had second trimester LC had normal deliveries at term. Of the 43 published cases of second trimester LC followed to delivery, 39 ended in uncomplicated, full-term deliveries. Three of four second trimester cases at one institution had spontaneous abortions. None of our patients underwent LC in the third trimester. Of the 12 published cases of third trimester LC followed to delivery, one had preterm delivery. Third trimester OC is reported to have a 40 percent rate of preterm delivery. There were no intraoperative cholangiograms (IOC), prophylactic or postoperative use of tocolytics, or intraoperative fetal monitoring in our series. We added six cases of LC during pregnancy to the previously reported 105 cases. The successful outcome in all trimesters suggests that LC is a safe procedure throughout pregnancy; however, surgery in the second trimester is preferable. Compared with OC, there is a decreased risk of spontaneous abortion in the first trimester and preterm labor in the third trimester. PMID- 9751940 TI - Nutritional and antimicrobial interventions to prevent preterm birth: an overview of randomized controlled trials. AB - The study was conducted to assess the effectiveness of interventions for the prevention or treatment of nutritional and infectious disorders during pregnancy on preterm birth rates. Cochrane systematic reviews or any other more up-to-date systematic review of antimicrobial and nutritional interventions were sought. Electronic searches of the Cochrane Controlled Trials Register were carried out to identify any trials published since the most recent update of the systematic review. Also, authors of Cochrane systematic reviews, which have not been updated recently, were contacted regarding new information. Systematic reviews of nutritional and antimicrobial interventions during pregnancy, reporting preterm delivery rates (delivery before 37 weeks) and "prematurity" (including low birth weight) either as primary or secondary outcomes, were included. General interventions without a specific nutritional supplementation or antimicrobial component were not considered for inclusion. Interventions to stop labor or prolong pregnancy after a diagnosis of preterm labor were excluded. For each systematic review, data on preterm delivery rate by intervention group was obtained. The total number of trials in the review, number of trials reporting preterm birth as an outcome, number of participants and events have been systematically extracted. Eighteen systematic reviews (10 nutritional and 8 antimicrobial) were considered. Our results indicated that, overall, the treatment of asymptomatic bacteriuria reduces the incidence of preterm birth or low birth weight (< 2500 gm) (typical relative risk (RR): 0.67; 95 percent confidence interval (CI): 0.52-0.85). The protective effect of treating asymptomatic bacteriuria for preterm delivery persisted when only the three trials reporting preterm delivery (< 37 weeks) were included in the meta-analysis (typical RR: 0.53, 95 percent CI: 0.33-0.86). Routine iron supplementation prevents maternal anemia and one trial comparing routine versus selective iron supplementation showed a statistically nonsignificant reduction in preterm birth. Zinc, magnesium, and fish oil supplementations show promising results in reducing preterm birth, but the evidence is not strong. Calcium supplementation remains controversial, although there was a statistically significant reduction in preterm delivery in the subgroup of women at high risk of developing hypertension during pregnancy. Two trials with use of metronidazole (alone or with erythromycin) showed a reduction in preterm delivery in women who were at a high risk of preterm delivery and had bacterial vaginosis at screening. We have concluded that asymptomatic bacteriuria should be screened and treated in all settings that offer antenatal care. Single dose treatment seems to be as effective as longer (4-7 days) treatment, although this needs to be confirmed by a large, methodologically rigorous trial. There are a number of promising interventions such as calcium supplementation in women with low calcium intake, iron, zinc, magnesium, and fish oil supplementation, and treatment of bacterial vaginosis in women at high risk of preterm delivery that need additional research to determine a possible role for prevention of preterm delivery. PMID- 9751941 TI - Metabolism of ethanol and some associated adverse effects on the liver and the stomach. AB - Current knowledge of alcohol oxidation and its effects on hepatic metabolism and its toxicity are summarized. This includes an evaluation of the relationship of the level of consumption to its interaction with nutrients (especially retinoids, carotenoids, and folate) and the development of various stages of liver disease. Ethanol metabolism in the stomach and its link to pathology and Helicobacter pylori is reviewed. Promising therapeutic approaches evolving from newly gained insight in the pathogenesis of medical complications of alcoholism are outlined. At present, the established approach for the prevention and treatment of alcoholism are outlined. At present, the established approach for the prevention and treatment of alcoholic liver injury is to control alcohol abuse, with the judicial application of selective antioxidant therapy, instituted at early stages, prior to the social or medical disintegration of the patient, and associated with antiinflammatory agents at the acute phase of alcoholic hepatitis. In addition, effective antifibrotic therapy may soon become available. PMID- 9751942 TI - Alcohol and the pancreas. AB - Alcoholic pancreatitis may be one of the most serious adverse consequences of alcohol abuse. Its diagnosis, as it has for many years, depends primarily on clinical acumen in interpreting properly the symptoms and signs of abdominal distress, buttressed by elevated pancreatic enzymes (amylase and lipase). More recently, the use of computerized tomography (CT) in selected situations has been both of confirmatory and prognostic value. Severity of abnormality by CT correlates reasonably well with a variety of clinical-laboratory clusters (APACHE system, Ranson's criteria, etc.) and aids in therapy. The pathogenesis of alcoholic pancreatitis is not fully defined. The ultimate picture is one of tissue autolysis by activated proteolytic enzymes. The triggers for such activation, however, are still not known. They are represented by three main theories: (1) large duct obstruction and/or increased permeability relative to pancreatic secretion, (2) small duct obstruction due to proteinaceous precipitates, and (3) a direct toxic-metabolic effect of ethanol on pancreatic acinar cells. While not mutually exclusive, we favor the last hypothesis as being most consistent with the effects of ethanol on other organ systems. The direct effects of ethanol and/or its metabolites may be mediated, at least in part, via oxidative stress or the generation of fatty acid ethyl esters. Autolysis (regardless of proximate mechanism(s)) leads to inflammation likely mediated via release of various cytokines. It also should be appreciated that "acute" pancreatitis (the topic of this chapter) likely represents an acute process within a chronic pancreatic exposure and injury from alcoholic abuse. The key question of why pancreatitis develops in only a small number of alcohol abusers is not resolved. Therapy depends on the severity of alcoholic pancreatitis, which is defined by clinical-laboratory and often CT criteria. Mild pancreatitis usually resolves acutely with alcohol abstention and supportive therapy. Severe pancreatitis has a significant morbidity and mortality, mainly related to the degree of pancreatic necrosis and infection. It requires meticulous combined medical-surgical care. PMID- 9751943 TI - Alcohol and cancer. AB - A great number of epidemiological data have identified chronic alcohol consumption as a significant risk factor for upper alimentary tract cancer, including cancer of the oropharynx, larynx, and the esophagus, and for the liver. In contrast to those organs, the risk by which alcohol consumption increases cancer in the large intestine and in the breast is much smaller. However, although the risk is lower, carcinogenesis can be enhanced with relatively low daily doses of ethanol. Considering the high prevalence of these tumors, even a small increase in cancer risk is of great importance, especially in those individuals who exhibit a higher risk for other reasons. The epidemiological data on alcohol and other organ cancers are controversial and there is at present not enough evidence for a significant association. Although the exact mechanisms by which chronic alcohol ingestion stimulates carcinogenesis are not known, experimental studies in animals support the concept that ethanol is not a carcinogen, but under certain experimental conditions is a cocarcinogen and/or (especially in the liver) a tumor promoter. The metabolism of ethanol leads to the generation of acetaldehyde and free radicals. These highly reactive compounds bind rapidly to cell constituents and possibly to DNA. Acetaldehyde decreases DNA repair mechanisms and the methylation of cytosine in DNA. It also traps glutathione, an important peptide in detoxification. Furthermore, it leads to chromosomal aberrations and seems to be associated with tissue damage and secondary compensatory hyperregeneration. More recently, the finding of considerable production of acetaldehyde by gastrointestinal bacteria was reported. Other mechanisms by which alcohol stimulates carcinogenesis include the induction of cytochrome P4502E1, associated with an enhanced activation of various procarcinogens present in alcoholic beverages, in association with tobacco smoke and in diets, a change in the metabolism and distribution of carcinogens, alterations in cell cycle behavior such as cell cycle duration leading to hyperregeneration, nutritional deficiencies such as methyl, vitamin A, folate, pyrridoxalphosphate, zinc and selenium deficiency, and alterations of the immune system, eventually resulting in an increased susceptibility to certain viral infections such as hepatitis B virus and hepatitis C virus. In addition, local mechanisms in the upper gastrointestinal tract and in the rectum may be of particular importance. Such mechanisms lead to tissue injury such as cirrhosis of the liver, a major prerequisite for hepatocellular carcinoma. Thus, all these mechanisms, functioning in concert, actively modulate carcinogenesis, leading to its stimulation. PMID- 9751944 TI - Alcohol and lipids. AB - Alcoholic fatty liver and hyperlipemia result from the interaction of ethanol and its oxidation products with hepatic lipid metabolism. An early target of ethanol toxicity is mitochondrial fatty acid oxidation. Acetaldehyde and reactive oxygen species have been incriminated in the pathogenesis of the mitochondrial injury. Microsomal changes offset deleterious accumulation of fatty acids, leading to enhanced formation of triacylglycerols, which are partly secreted into the plasma and partly accumulate in the liver. However, this compensatory mechanism fades with progression of the liver injury, whereas the production of toxic metabolites increases, exacerbating the lesions and promoting fibrogenesis. The early presence of these changes confers to the fatty liver a worse prognosis than previously thought. Alcoholic hyperlipemia results primarily from increased hepatic secretion of very-low-density lipoprotein and secondarily from impairment in the removal of triacylglycerol-rich lipoproteins from the plasma. Hyperlipemia tends to disappear because of enhanced lipolytic activity and aggravation of the liver injury. With moderate alcohol consumption, the increase in high-density lipoprotein becomes the predominant feature. Its mechanism is multifactorial (increased hepatic secretion and increased extrahepatic formation as well as decreased removal) and explains part of the enhanced cholesterol transport from tissues to bile. These changes contribute to, but do not fully account for, the effects on atherosclerosis and/or coronary heart disease attributed to moderate drinking. PMID- 9751945 TI - Cardiovascular effects of alcohol. AB - The ingestion of one or two alcoholic drinks can affect heart rate, blood pressure, cardiac output, myocardial contractility, and regional blood flow. These actions generally are not clinically important. In the presence of cardiovascular disease, however, even such small quantities of alcohol might result in transient unfavorable hemodynamic changes. Moreover, alcohol abuse can produce cardiac arrhythmias, hypertension, cardiomyopathy, stroke, and even sudden death. In contrast, moderate alcohol use produces changes that have an overall favorable effect on atherosclerotic-related vascular diseases. Because cardiovascular disease due to atherosclerosis is the leading cause of death in Western society, this desirable effect of alcohol use outweighs its detrimental actions, resulting in favorable findings in population studies. Nevertheless, the body of evidence argues against encouraging alcohol use for its cardiovascular effects. PMID- 9751946 TI - Mechanisms of alcohol craving and their clinical implications. AB - Craving for alcohol is frequently given as a reason for drinking and is often used as a surrogate measure in studies of alcoholism and its treatment. Despite this wide use, there is little consensus on what craving for alcohol means, the best way to measure it, what mechanism accounts for the urge to drink, or what is its true relationship to alcohol use. This chapter reviews theoretical and measurement issues about the possible mechanisms involved in craving for alcohol and the clinical implications of evidence supporting them. Until recently, most instruments for assessing craving assumed it was a univariate construct and usually contained only one or a few items. Several multi-item and multidimensional rating instruments have now been developed that offer the promise of more useful assessment of clinically relevant behavior. Most models of craving have assumed that a consistent and positive relationship exists between craving and drinking. The incentive sensitization model and the cognitive theory of drug use and drug urges may account better than the older models for the frequent clinical observation of a dissociation between craving and drinking. However, no single model or theory of craving accounts for the wide variation in findings reviewed here, suggesting that multiple mechanisms may be involved. A comprehensive, multidisciplinary approach is necessary to elucidate the nature of craving for alcohol and its implications for pharmacological and psychosocial treatment of alcoholism. PMID- 9751947 TI - A review of the effects of moderate alcohol intake on psychiatric and sleep disorders. AB - In this chapter we discuss the effects of moderate ethanol consumption on the treatment of psychiatric and sleep disorders. A review of the literature on the interactions of ethanol with neurotransmitters and psychotropic medications suggests that although ethanol affects the clinical course of psychiatric and sleep disorders by different mechanisms, it does so principally through perturbations it causes in the balance of central nervous system neurotransmitter systems, which may modify the clinical course of primary psychiatric and sleep disorders and undermine the therapeutic response to psychotropic medications. Neurotransmitter responses may also be manifested clinically by rebound phenomena, akin to a subsyndromal withdrawal, which affect sleep and precipitate anxiety and mood symptoms. In addition, ethanol also modifies the clearance and disposition of a variety of psychotropic metabolites and interferes with their clinical effectiveness. We recommend that most psychiatric patients, and all patients with sleep disorders, should abstain from even moderate ethanol use, as this may adversely affect their clinical course and response to treatment. PMID- 9751948 TI - Executive cognitive functioning in alcohol use disorders. AB - Executive cognitive functioning (ECF) has been identified as an important determinant in the etiology of alcoholism. ECF represents a "higher-order" cognitive construct involved in the self-regulation of goal-directed behavior. The prefrontal cortex and its subcortical connections represent the primary neurological substrate that subserves ECF. Both alcoholics and individuals at high risk for alcoholism exhibit a mild dysfunction in ECF. However, this deficit appears to be significantly stronger in alcoholics with a comorbid diagnosis of an antisocial personality disorder. Individuals with other disorders that are also highly comorbid with alcoholism, such as attention deficit hyperactivity disorder and conduct disorder, also demonstrate deficits in ECF. As such, compromised ECF may not be specific to alcoholism, but instead, might be a potential underlying etiologic substrate for a number of disorders of behavioral excess-disinhibition. Subsequent to reviewing the literature implicating ECF deficits in alcoholism and comorbid disorders, the authors present a heuristic cognitive-neurobehavioral model of alcoholism implicating the frontostriatal system. Finally, recommendations for the prevention and treatment of alcoholism, based on this model, are discussed. PMID- 9751949 TI - Brain imaging. Functional consequences of ethanol in the central nervous system. AB - In recent years, sophisticated methods have been developed to view structure and function within the living brain. Functional imaging methods are used to visualize dynamic chemical processes that are linked to brain activity. Increased neural activity, for example, leads to greater glucose and oxygen consumption and greater regional rates of blood flow to meet elevated energy demands. Mapping these changes provides quantitative visual descriptions of localized changes in brain activity that result from behavioral or pharmacological manipulations. This chapter first describes several current methods and how they are used to study the effects of alcohol on brain function. In the second part, the effects of acute intoxication are discussed with emphasis on the complex nature of alcohol's effects in the central nervous system, which depend on dose, time since administration, and environmental context. In the final part, the functional consequences of long-term exposure to alcohol as well as diseases associated with chronic alcoholism are reviewed. PMID- 9751950 TI - Complications of severe mental illness related to alcohol and drug use disorders. AB - In this chapter we review research on the relationships between substance use disorder and 11 domains of adjustment for people with severe mental illness. Studies are divided into correlational research and prospective, longitudinal research, with greater weight given to those in the latter category. The weight of the evidence indicates that substance abuse severely complicates severe mental illness in the following domains: relapse of psychiatric illness, hospitalization, disruptive behavior, familial problems, residential instability, decreased functional status, HIV infection, and medication noncompliance. We discuss the limits of causal inference in these studies and the possible mechanisms that relate substance abuse to various complications. PMID- 9751951 TI - Economic costs of alcohol abuse and alcoholism. AB - The economic cost to society from alcohol abuse and alcoholism in the United States was an estimated $148 billion in 1992. When adjusted for inflation and population, the estimates are generally comparable with cost estimates produced over the past 20 years. The current estimates are significantly greater than the most recent detailed estimates developed for 1985--about 42% above increases due to population growth and inflation. Between 1985 and 1992, inflation accounted for about 37.5% and population growth for 7.1% increases. Changes in prevalence have been associated with a modest reduction in alcohol costs. Though crime rates did not materially change over this period, criminal justice expenditures more than doubled overall, even after adjustment for price increases. The balance of changes are due to new findings and/or methodology indicating larger impacts than previously estimated. It is estimated that 45.1% of costs are borne by alcohol abusers and/or members of their households, 38.6% are borne by government, 10.2% by private insurance, and 6.0% by victims of alcohol-related trauma (motor vehicle crashes plus crime). The costs staying in the household of the abusers may be materially incident on persons other than the abuser, e.g., spouses, children. PMID- 9751952 TI - The effects of price on the consequences of alcohol use and abuse. AB - Economists have examined the impact of alcohol prices on various outcomes related to alcohol consumption, including nonfatal and fatal motor vehicle accidents, other accidents, liver cirrhosis, and other alcohol-related mortality, crime, and education attainment. Price, in the context of this research, includes not only the monetary price of alcoholic beverages, but also a wide variety of other "costs" of drinking and heavy drinking, including the time spent obtaining alcoholic beverages and the legal costs associated with drinking and related behavior. This research clearly demonstrates that increases in the monetary prices of alcoholic beverages, which could be achieved by increasing taxes on alcohol, can significantly reduce many of the problems associated with alcohol use and abuse. In addition, control policies that raise other "costs" of drinking, including reduced availability of alcoholic beverages, higher legal drinking ages, and others, are also effective in reducing the consequences of alcohol use and abuse. PMID- 9751953 TI - Drinking, problem drinking, and productivity. AB - This chapter surveys and critiques the recent economic literature dealing with the relationships between labor market productivity and alcohol use and misuse. The focus here is twofold. First is to present and discuss the relevant conceptual issues that must be appreciated in assessing such relationships. Second is to summarize and assess the empirical findings that have been offered in the literature. PMID- 9751954 TI - The cost offsets of alcoholism treatment. AB - While the effectiveness of alcoholism treatment is an important concern in alcohol research, the cost of such treatment and its benefits are also important research matters. There is substantial research that examines the possible benefits of alcoholism treatment in reducing the cost of all medical care, including the cost of alcoholism treatment itself. This is referred to as cost offsets. This chapter reviews the research evidence of alcoholism treatment cost offset, that is, the ability of alcoholism treatment to reduce the cost of medical care of persons participating in such treatment. The chapter gives an overview summary of the cost offset findings for alcoholism treatment and concludes with an identification of future research needs and opportunities, especially surrounding the popular increase in the use of managed care. PMID- 9751955 TI - Experience with the alcohol use disorders identification test (AUDIT) in Mexico. AB - This chapter describes the development of the Alcohol Use Disorders Identification Test (AUDIT) among various Mexican populations, the evaluations that followed the World Health Organization international research project from where this screening instrument was derived, its use in nonclinical settings, modifications introduced in its wording, the development of a short version, and validity and reliability tests. It also describes rates of hazardous, harmful, and dependent drinkers and biobehavioral consequences of abuse among various Mexican populations. Data drawn from different samples showed adequate levels of specificity and sensitivity. Findings from general population samples confirmed previous observations in general practice: That the AUDIT could capture not only regular consumption at hazardous levels, but also episodic heavy drinking. Data from an International Labor Office/World Health Organization project on model programs for alcohol prevention in the workplace showed that it was possible to derive a short version, easily used for intervention programs, that differentiated categories of drinkers at various risk levels. Rates of problem drinkers in clinical samples varied between 28 and 43% for males and 3.6 and 4.8% among females. Hazardous drinking varied between 0.7 and 15.5% among males and females in general populations and reached 44% in a sample of male workers; in clinical settings, harmful drinking ranged from 7 to 16% among males and dependence from 3 to 10%. PMID- 9751956 TI - Problems associated with hazardous and harmful alcohol consumption in Mexico. AB - This chapter presents research findings from a collaborative project between Mexican investigators from the Mexican Institute of Psychiatry and the World Health Organization on the identification and treatment of harmful and hazardous drinking. A sample of 189 individuals who met criteria for hazardous drinking was selected for the study after screening 2319 outpatients attending clinics in two general hospitals in Mexico City. We present here the characteristics of this sample along dimensions that include alcohol related problems, history of trauma, alcohol dependence scores and family history of alcoholism. We rated, utilizing structures interviews, situations that place these individuals at risk of drinking. The possibility of constructing a typology of harmful and hazardous drinking was also explored. The significance of the findings of this investigation for health care clinicians is discussed. PMID- 9751957 TI - Sanctification of "the accursed". Drinking habits of the French existentialists in the 1940s (a case study). AB - The chapter deals with the drinking habits of the French existentialists during and after World War II (roughly from 1943 to 1948). It attempts to show that the phenomenon cannot be understood separately from their lifestyle as a whole, which in this case (as I claim) is primarily manifested through a certain (mythical) structure of meanings related to the category of the sacred. Jean-Paul Sartre's position as the leading intellectual figure of the time is also to be seen as a result of his ambiguous reputation as a "prophet" and a "criminal" in the yellow press. What is involved is a single mythical structure, where "decadent" life is precisely one aspect of sanctification. On the other hand, Sartre's "bohemian lifestyle" and his desire to break bourgeois habits can be seen as a variant of the bourgeois myth of the artistic lifestyle created in the 19th century. From this angle, existentialism can in a certain sense be considered the last hybrid expression of the "transgressive" myth of rebellion which this sort of lifestyle had crystallized. PMID- 9751958 TI - Cocaine metabolism in humans after use of alcohol. Clinical and research implications. AB - The simultaneous administration of cocaine and alcohol implies a pharmacological interaction at pharmacodynamic and pharmacokinetic levels. The latter involves an alteration of cocaine kinetics and metabolism, as well as the biosynthesis of newly active metabolites, such as cocaethylene. Cocaethylene is metabolized along the same pathways as cocaine. Its detection in biological samples indicates the combined consumption of cocaine and alcohol. From epidemiological and toxicological data, it has been suggested that the combination of alcohol and cocaine produces an increased toxicity in addition to behavioral changes. There has been some debate regarding the contribution of cocaethylene to this rise of toxicity. Its pharmacological and toxicological profile is very similar to cocaine. During the interaction of both substances, the rise in cocaine plasma concentrations can explain many of cardiovascular and behavioral effects observed. The contribution of cocaethylene to the interaction is probably minor; its effects are likely additive to those of cocaine. Perhaps its longer elimination half-life can help in understanding long-lasting effects of the alcohol-cocaine combination. PMID- 9751959 TI - Interrelationship between alcohol intake, hepatitis C, liver cirrhosis, and hepatocellular carcinoma. AB - The discovery of a cDNA clone of hepatitis C virus (HCV) genome in 1989 has resulted in numerous reports of high rates of the prevalence of HCV antibody in patients with alcoholic liver disease, in particular, alcoholic liver cirrhosis and hepatocellular carcinoma. Thus, the interaction between alcohol intake and HCV infection has become of great importance. In terms of the effect of alcohol on HCV-RNA levels, the data are controversial; in some reports, alcohol increases HCV-RNA levels, and in the other reports it does not. There are several reports suggesting the possibility of an elevated quasi-species of hypervariable region 1 in the HCV genome caused by alcohol drinking. Recent studies have documented that alcohol intake exaggerates the responsiveness of interferon therapy for chronic hepatitis C; however, its mechanism is still obscure. Several studies have suggested the promoting effect of alcohol on the development of hepatocellular carcinoma in type C liver cirrhosis, which has been similarly observed in type B chronic liver disease, whereas its mechanism is limited to speculations. PMID- 9751960 TI - [The principles of orthodoxy and the spiritual rebirth of the Russian people]. PMID- 9751961 TI - [The concept of the clinical use of serotonin adipinate in treating surgical patients]. AB - The influence of serotonin adipinate on the mechanisms of development of tissue hypoxia, disturbances of the microcirculatory bed and smooth muscles before, during and after operative interventions was studied in experiments and clinical practice. The syndrome of serotonin insufficiency is described including smooth muscle insufficiency which can be abolished by serotonin adipinate administered at the early postoperative period irrespective of the cause of surgery, age and sex of the patient. Good results were obtained in treatment of patients after operations on the liver, pancrease, heart, in patients with the diabetic foot and in other diseases. No complications or lethal outcomes resulting from using serotonin adipinate were noted. PMID- 9751962 TI - [Means for the safe surgical treatment of patients with thyroid diseases]. AB - An analysis of a 35-years experience with organ-saving surgical treatment of 13,751 patients with diseases of the thyroid gland has been made. The basic principles of prevention of recurrent diseases and specific postoperative complications are formulated. An original technique of operations resulting in less frequent injuries of the recurrent nerves, hypoparathyrosis and postoperative lethality after surgical treatment of patients with benign diseases and carcinoma of the thyroid gland has been developed. PMID- 9751963 TI - [The single-row suture in the pyloroplasty of patients with perforated pyloroduodenal ulcers]. AB - The authors made an analysis of the immediate and long-term results of truncal vagotomy for perforating gastroduodenal ulcers in accordance with the kind of the suture in operation of stomach drainage. In 282 patients pyloroplasty was performed with placing a two-row suture, in 239 patients a one-row suture was used. Lethality among the patients with the two-row sutures was 2.1% in those with the one-row suture--0.4%. The placing of one-row sutures was followed by a relatively smaller amount of complications at the early postoperative period. An analysis of long-term results has shown that the one-row suture gives much greater amount of excellent results as compared with the two-row suture. The authors recommend to use the one-row suture in all the cases of perforating pyloroduodenal ulcers irrespective of the phase of intraperitoneal inflammation. PMID- 9751964 TI - [2-Component silicone compounds in the emergency surgery of gastrointestinal hemorrhages]. AB - Based upon their clinico-experimental investigations the authors prove advantages of the two-component silicon compositions over oil preparations for the arrest of gastro-intestinal bleedings. Such compositions are recommended for patients whose treatment by traditional methods of hemostasis proved to be ineffective due to some causes which is often observed in elderly and senile patients. PMID- 9751965 TI - [New methods for assessing the severity of endogenous intoxication in surgical patients]. AB - New methods (a reticulocytic test, inhibition of erythrocyte sedimentation, registration of the frog's heart rate) were used in studying the toxicity of the serum, blood plasma and urine of surgical patients. The most informative methods for the assessment of the toxemia degree are thought to be the reticulocytic test and registration of the frog's heart rate. The maximum time for the determination of endotoxicity is 15 minutes. The pathological process severity can be estimated not only by the toxemia degree but also by its dynamics. The determination of endotoxicity is of value for choosing the methods for the individual detoxication therapy. PMID- 9751967 TI - [The characteristics of the surgical procedure in isolated gunshot wounds of the leg in peacetime]. AB - The article presents results of treatment of 20 people with gunshot wounds of the leg for the period from 1994 till 1996. The authors used the methods directed to treatment of the traumatic disease, correction of the regional blood circulation and microcirculation, firm fixation of the fractures, adequate drainage and fasciotomy fulfilled if indicated. PMID- 9751966 TI - [The effect of pancreatic islet cell implantation on the course of diabetic nephropathy]. AB - The influence of the implanted allo- and xenocultures of the pancreas islet cells upon the course of nephropathy was studied in 61 patients with insulin-dependent diabetes mellitus. These cells were found to have a positive influence upon the functional state of the kidneys in patients with diabetic nephropathy especially at its initial stage. PMID- 9751968 TI - [Heterodelphus--a rare form of twin developmental defect]. AB - The authors saw a rare developmental twin anomaly--heterodelphus. The parasite was an organism devoid of the head, fixed with its thoracic wall an epigastric area to the front wall and the epigastric area of the autosite. Respiration and palpitation of the parasite was not auscultated. The abnormal twins had a single umbilical cord. The surgical division of the conjoined twins was performed in 4 months. The operation and operative findings are described. The postoperative period was hard. At the present time the boy's state is good, he feels well and develops in accordance with his age. PMID- 9751969 TI - [The advantage of the laparoscopic method of examining patients with nonpalpable testes]. AB - The authors compared the diagnostical value of traditional operations with the inguinal access and a new method using laparoscopy for treatment of patients with testis aplasia. The laparoscopic method of examination was found to be preferable since it allows the correct diagnosis to be made in 96% of children with cryptorchism. PMID- 9751970 TI - [Aortic valve prosthesis implantation from a mini approach]. AB - The authors share their experiences with performing the first in Russia operation of making the aorta valve prosthetics from a mini-access--transverse sternotomy. It is concluded that operations through such accesses are safe for the patient and relatively easy for the surgeon. Its main advantage consists in quick surgical rehabilitation of the patient and good cosmetic effect. PMID- 9751971 TI - [Surgical methods for correcting partial anomalous drainage of the pulmonary veins during artificial circulation]. AB - The authors share their experiences with surgical treatment of 42 patients with different anatomical variants of the partial anomalous drainage of the pulmonary veins. Specific features of the surgical strategy are described depending on the level of confluence of the pulmonary veins, especially if the mouths of the abnormally falling pulmonary veins and the interatrial septum defect are far from each other. Good immediate and nearest results were obtained. PMID- 9751972 TI - [A method of phalloplasty using a non-free skin-fat flap from the thigh]. AB - The authors propose a method of phalloplasty with a full-thickness flap on the vasculo-nervous pedicle taken from the femur. The method of raising such a flap and the technique of its displacement to the area of the pubic articulation if described. This method of phalloplasty was used in 10 patients aged 19-36 years. In all of them full taking of the transplant was noted. Good cosmetic and functional results were obtained. PMID- 9751973 TI - [Regional arterial infusion in patients after reconstructive-restorative operations of the hip joint]. AB - The effectiveness of the regional arterial infusion in prophylaxis of complications was studied in 14 patients after reconstructive-restorative operations upon the hip joint. The method and program of the complex infusion therapy are proposed which facilitate the correction of disturbed microcirculation and oxygen regimen in the tissues after operation (trauma) on the hip joint. PMID- 9751974 TI - [The use of laparostomy and drainage in treating peritonitis]. AB - The laparostomic method is described in detail when used in treatment of peritonitis. Treatment of 6 patients with diffuse peritonitis by this method gave good effect. PMID- 9751975 TI - [Rare causes of the dysfunction of mechanical bicuspid heart valves]. AB - The authors share the observations from their cardiosurgical practice showing that one of the causes of sudden death or of rapid worsening of the state of patients with implanted bicuspid artificial valves might be their dysfunction resulting both from breakage of the hinged mechanisms of the valve with its dislocation and migration and from the disturbed mobility of the valve cusp due to a thrombus formed in the atrium. The observations show that the "ideal" construction of the valve is to be created. PMID- 9751976 TI - [The late results of combined surgical treatment in peptic ulcer patients]. AB - Long-term results (for 10 and more years) were analyzed in 661 of 693 patients operated upon for ulcer disease of the stomach and duodenum. The complex antiulcerous treatment before operation, immediately after it, at the nearest and remote periods, the sanatorium and resort treatment were given to 377 patients while 316 patients were treated by the generally accepted methods only, without a sanatorium rehabilitation. The best results were obtained after all kinds of operations (resection of the stomach, vagotomy with resection of the stomach, truncal and selective vagotomy with drainage operations, selective proximal vagotomy) in patients treated by complex methods with the sanatorium rehabilitation. The effect was most pronounced in patients after organ-saving operations and in those having a short ulcer history. PMID- 9751977 TI - [The surgical procedure in the Mallory-Weiss syndrome]. AB - The authors dealt with treatment of 112 patients aged 27-70 years with the Mallory-Weiss syndrome. The diagnosis was confirmed by esophagogastroduodenoscopy. Diathermocoagulation was used in order to arrest bleeding. In profuse bleeding the margins of the mucosa fissures were first infiltrated with a solution of adrenaline. The Blakemore [correction of Bleikmorr] probe compression method was also used. Organ-saving operations were performed for continuing and recurrent bleedings. Two elderly patients with severe coexistent disease died. The authors consider that patients with the Mallory-Weiss syndrome must be treated by conservative methods. Operations for disruptions of the esophagus mucosa and acute blood loss will entail great risk. PMID- 9751978 TI - [Laparoscopy in perforated gastroduodenal ulcers]. AB - The authors have performed operations on 32 patients with perforated ulcers of the duodenum and 7 patients with perforated ulcers of the stomach. The diameter of the perforations was 2-8 mm. In 10 of the 39 patients the perforation defects could not be sutured by the laparoscopic method. The authors consider that of great significance for the decision to make laparoscopic operations was the diagnosis of peritonitis, size and localization of the perforation, the surgeon's experience with endoscopic operating. The technique of laparoscopic suturing the perforations is described. Special attention is paid to the special disposition of the surgeon and his assistant at the operating table. PMID- 9751979 TI - [Surgical care in duodenal injuries]. AB - The authors observed 40 patients with an injury of the duodenum. Isolated injuries of the duodenum were diagnosed in 10 patients with a closed trauma and in 3 patients with penetrating wounds of the abdomen. Most frequent are injuries of the duodenum in combination with traumas of other organs of the abdominal cavity. Patients with injuries of the duodenum can be treated by surgical methods only. Methods of revision of the duodenum during laparotomy are described as well as the expedient surgical policy for injuries of the duodenum and postoperative complications. Clinical observations are presented. A conclusion is made that at the acute period of trauma in patients with multiple injuries of organs of the abdominal cavity and severe blood loss the operation of choice must consist in suturing the wound of the duodenum with the reinforcement of the suture with a portion of the greater omentum, decompression of the gut with a nasointestinal probe established distally to the suture on the duodenum and draining the retroperitoneal hematoma. PMID- 9751980 TI - [Ultrasonic diagnosis in the surgical treatment of angiodysplasias of the lower extremities]. AB - An experience with examination and surgical treatment of 156 patients with angiodysplasia of lower extremities shows that the ultrasonic location and dopplerography are highly informative methods of diagnosis of this disease. The orthostatic probe (the Valsalva test effect) allows the spread of angiodysplasia (suprafascial or subfascial) to be determined. The adequate strategy and volume of surgery can be chosen correctly. These methods of examination being noninvasive and easy makes them helpful for the objective assessment of the effectiveness of the surgical procedures and control of further treatment. PMID- 9751982 TI - [Odontogenic total mediastinitis complicated by an arrosive hemorrhage from the major vessels of the neck]. PMID- 9751981 TI - [The parenteral administration of terridecase in the treatment of suppurative inflammatory diseases of the hand]. AB - Patients with purulent inflammatory diseases were treated by parenteral administration of terrydecaza (polyglucin-immobilised terrylythin). It was noted that injections of terrydecaza facilitated the penetration of antibiotics to the purulent focus. The terms of cleansing the purulent wounds and transition of the inflammatory process to the second stage were 2-3 days shorter. The microcirculation was improved by the daily thermographic monitoring method. The increased penetration through the biological membranes was confirmed by higher concentration of middle weight molecules in plasma of the venous blood flowing off from the purulent focus. PMID- 9751983 TI - [Phlegmon of the stomach simulating a perforated ulcer]. PMID- 9751984 TI - [The efficacy of the surgical and combined treatment of complicated forms of malignant lymphomas of the stomach]. PMID- 9751985 TI - [Perforated duodenal and stomach ulcers in complete situs inversus viscerum]. PMID- 9751986 TI - [The sequelae of the laparoscopic suturing of a perforated duodenal ulcer]. PMID- 9751987 TI - [Right-sided hemicolectomy combined with gastropancreatoduodenal resection in locally disseminated colonic cancer]. PMID- 9751988 TI - [Adenoma of the liver in a young woman]. PMID- 9751989 TI - [Precision terminolateral hepaticoduodenal anastomosis in damage to a small diameter hepaticocholedochus]. PMID- 9751990 TI - [Adenomatous hyperplasia of the common hepatic duct as the cause of high biliary duct stricture]. PMID- 9751991 TI - [Sepsis. The importance of unified concepts (apropos the articles by M. V. Grinev and M. I. Gromov "Sepsis. The polemical aspects of the problem" and I. A. Eriukhin and S. A. Shliapnikov "Generalized forms of the inflammatory reaction and of surgical infection. Current problems in the terminology and delimitation of concepts")]. PMID- 9751992 TI - [Abdominal sepsis (based on materials from a roundtable)]. PMID- 9751993 TI - [The use of allografts in the surgical treatment of patients with infected vascular prostheses]. AB - The article is devoted to analysis of surgical treatment of patients with such a dangerous complication as infection of the vascular prosthesis. The authors have performed 25 operations for substituting the infected prostheses for allografts. In 18 patients the infected prostheses were located in the aorto-iliac segment, in 5 patients in the femoro-popliteal segment, 1 patient had it in the aorta, and 1 in the subclavian segment. The prostheses were removed and a simultaneous revascularization was made. The authors make a conclusion that it is very expedient to use alloprostheses as a protective measure against persistent or recidivating infections. The allografts have both early and long-term resistance to infection. PMID- 9751994 TI - [Wounds of the heart and their surgical treatment. 1927 (excerpts from the book)]. PMID- 9751995 TI - Hydrogen bonding geometry of a protein-bound carbohydrate from water exchange mediated cross-relaxation. AB - We present heteronuclear two-dimensional methods for the analysis of the geometry of exchangeable protons on a protein-bound carbohydrate. By using a water selective NOESY-HSQC, we observed cross-relaxation between carbohydrate hydroxyl protons and non-exchangeable ring protons in the complex of [13C6]-alpha-methyl-D mannopyranoside with recombinant rat mannose binding protein. Using a simple kinetic model, we were able to explain the differences in the initial slopes of the resulting cross-relaxation buildup curves in terms of the geometry of the hydroxyl protons in the bound state. The hydroxyl rotamers consistent with our cross-relaxation data fit very well with predictions based on the crystal structure of MBP bound to a mannose-rich oligosaccharide. These methods should be applicable to other systems where both ligand exchange and water exchange are fast relative to the rate of cross-relaxation. PMID- 9751996 TI - Rotational-echo double-resonance in complex biopolymers: a study of Nephila clavipes dragline silk. AB - Rotational-Echo Double-Resonance (REDOR) NMR on strategically 13C and 15N labeled samples is used to study the conformation of the LGXQ (X = S, G, or N) motif in the major ampullate gland dragline silk from the spider Nephila clavipes. A method is described for calculating REDOR dephasing curves suitable for background subtractions, using probability distributions of nitrogen atoms surrounding a given carbon site, which are developed from coordinates in the Brookhaven Protein Data Bank. The validity of the method is established by comparison to dephasings observed from natural abundance 13C peaks for G and A. Straightforward fitting of universal REDOR dephasing curves to the background corrected peaks of interest provides results which are not self-consistent, and a more sophisticated analysis is developed which better accounts for 15N labels which have scrambled from the intended positions. While there is likely some heterogeneity in the structures formed by the LGXQ sequences, the data indicate that they all form compact turn-like structures. PMID- 9751997 TI - Nuclear magnetic resonance characterization of a paramagnetic DNA-drug complex with high spin cobalt; assignment of the 1H and 31P NMR spectra, and determination of electronic, spectroscopic and molecular properties. AB - The proton NMR spectrum of the ternary complex between the octamer duplex d(TTGGCCAA)2, two molecules of the drug chromomycin-A3, and a divalent cobalt ion has been assigned. Assignment procedures used standard two-dimensional techniques and relied upon the expected NOE contacts observed in the equivalent diamagnetic complex containing zinc. The magnetic susceptibility tensor for the cobalt was determined and used to calculate shifts for all nuclei, aiding in the assignment process and verification. Relaxation, susceptibility, temperature and field dependence studies of the paramagnetic spectrum enabled determination of electronic properties of the octahedral cobalt complex. The electronic relaxation tau(s) was determined to be 2.5 +/- 1.5 ps; the effective isotropic g value was found to be 2.6 +/- 0.2, indicating strong spin-orbit coupling. The magnetic susceptibility tensor was determined to be chi(xx) = 8.9 x 10(-3) cm3/mol, chi(yy) = 9.5 x 10(-3) cm3/mol, chi(zz) = 12.8 * 10(-3) cm3/mol. A tentative rotational correlation time of 8 ns was obtained for the complex. Both macroscopic and microscopic susceptibility measurements revealed deviations from Curie behavior over the temperature range accessible in the study. Non-selective relaxation rates were found to be inaccurate for defining distances from the metal center. However, pseudocontact shifts could be calculated with high accuracy using the dipolar shift equation. Isotropic hyperfine shifts were factored into contact and dipolar terms, revealing that the dipolar shift predominates and that contact shifts are relatively small. PMID- 9751999 TI - Practical model fitting approaches to the direct extraction of NMR parameters simultaneously from all dimensions of multidimensional NMR spectra. AB - A maximum likelihood (ML)-based approach has been established for the direct extraction of NMR parameters (e.g., frequency, amplitude, phase, and decay rate) simultaneously from all dimensions of a D-dimensional NMR spectrum. The approach, referred to here as HTFD-ML (hybrid time frequency domain maximum likelihood), constructs a time-domain model composed of a sum of exponentially-decaying sinusoidal signals. The apodized Fourier transform of this time-domain signal is a model spectrum that represents the 'best-fit' to the equivalent frequency domain data spectrum. The desired amplitude and frequency parameters can be extracted directly from the signal model constructed by the HTFD-ML algorithm. The HTFD-ML approach presented here, as embodied in the software package CHIFIT, is designed to meet the challenges posed by model fitting of D-dimensional NMR data sets, where each consists of many data points (10(8) is not uncommon) encoding information about numerous signals (up to 10(5) for a protein of moderate size) that exhibit spectral overlap. The suitability of the approach is demonstrated by its application to the concerted analysis of a series of ten 2D 1H-15N HSQC experiments measuring 15N T1 relaxation. In addition to demonstrating the practicality of performing maximum likelihood analysis on large, multidimensional NMR spectra, the results demonstrate that this parametric model fitting approach provides more accurate amplitude and frequency estimates than those obtained from conventional peak-based analysis of the FT spectrum. The improved performance of the model fitting approach derives from its ability to take into account the simultaneous contributions of all signals in a crowded spectral region (deconvolution) as well as to incorporate prior knowledge in constructing models to fit the data. PMID- 9751998 TI - Some NMR experiments and a structure determination employing a [15N,2H] enriched protein. AB - We present the results of studies of an aqueous sample of a highly [15N,2H] enriched protein, the SH3 domain from Fyn. Measurements of 1H relaxation and interactions between H2O solvent and exchangeable protons are given, as well as a method for increasing the effective longitudinal relaxation of solvent exchangeable proton resonances. The long-range isotope shifts are measured, for 1H and 15N, which arise due to perdeuteration. Simulations, which employed a 7 or 8 spin relaxation matrix analysis, were compared to the experimental data from a time series of 2D NOESY datasets for some resonances. The agreement between experiment and simulation suggest that, with this 1H dilute sample, relatively long mixing times (up to 1.2 s) can be used to detect specific dipolar interactions between amide protons up to about 7A apart. A set of 155 inter-amide NOEs and 7 side chain NOEs were thus identified in a series of 3D HSQC-NOESY-HSQC experiments. These data, alone and in combination with previously collected restraints, were used to calculate sets of structures using X-PLOR. These results are compared to the available X-ray and NMR structures of the Fyn SH3 domain. PMID- 9752000 TI - An asymmetric deuterium labeling strategy to identify interprotomer and intraprotomer NOEs in oligomeric proteins. AB - A major difficulty in determining the structure of an oligomeric protein by NMR is the problem of distinguishing inter- from intraprotomer NOEs. In order to address this issue in studies of the 27 kD compact trimeric domain of the MHC class II-associated invariant chain, we compared the 13C NOESY-HSQC spectrum of a uniformly 13C-labeled trimer with the spectrum of the same trimer labeled with 13C in only one protomer, and with deuterium in the other two protomers. The spectrum of the unmixed trimer included both inter- and intraprotomer NOEs while the spectrum of the mixed trimer included only intraprotomer peaks. NOEs clearly absent from the spectrum of the mixed trimer could be confidently assigned to interprotomer interactions. Asymmetrically labeled trimers were isolated by refolding a 13C-labeled shorter form of the protein with a 2H-labeled longer form, chromatographically purifying trimers with only one short chain, and then processing the trypsin to yield only protomers with the desired N- and C-termini. In contrast to earlier studies, in which statistical mixtures of differently labeled protomers were analyzed, our procedure generated only a well-defined 1:2 oligomer, and no other mixed oligomers were present. This increased the maximum possible concentration of NMR-active protomers and thus the sensitivity of the experiments. Related methods should be applicable to many oligomeric proteins, particularly those with slow protomer exchange rates. PMID- 9752001 TI - Identification of slow motions in the reduced recombinant high-potential iron sulfur protein I (HiPIP I) from Ectothiorhodospira halophila via 15N rotating frame NMR relaxation measurements. AB - Rotating-frame 15N relaxation rate (R1 rho) NMR experiments have been performed in order to study the dynamic behavior of the reduced recombinant high-potential iron-sulfur protein iso I (HiPIP I) from Ectothiorhodospira halophila, in the microsecond to ms time range. Measurements of R1 rho were performed as a function of the effective spinlock magnetic field amplitude by using both on and off resonance radio frequency irradiation. The two data sets provided consistent results and were fit globally in order to identify possible exchange processes in an external loop of the reduced HiPIP I. The loop consists of residues 43-45 and the correlation time of the exchange process was determined to be 50 +/- 8 microseconds for the backbone nitrogen of Gln 44. PMID- 9752002 TI - Rapid measurement of scalar three-bond 1HN-1H alpha spin coupling constants in 15N-labelled proteins. AB - Two new 2D NMR experiments, CT-HMQC-HA and CT-HMQC-HN, are proposed for the rapid measurement of homonuclear 3JHNH alpha coupling constants of uniformly 15N enriched proteins in solution. The experiments are based on the comparison of the signal intensities in a pair of constant-time [15N,1H]-HMQC spectra recorded with and without decoupling of the amide proton-alpha proton coupling. Experimental data recorded with the 78-residue N-terminal domain of the E. coli arginine repressor (ArgR-N) and with oxidized E. coli flavodoxin (176 residues) showed good agreement with 3JHNH alpha coupling constants obtained by fitting of the multiplet fine structure of the amide proton resonances or from a 3D HNHA-J experiment, respectively. Quantitative estimates for the effects from different relaxation rates of in-phase and antiphase magnetization are given. PMID- 9752004 TI - 1H, 13C, 15N resonance assignment of the 20 kDa double stranded RNA binding domain of PKR. PMID- 9752005 TI - 1H, 13C and 15N NMR resonance assignments of vaccinia glutaredoxin-1 in the fully reduced form. PMID- 9752006 TI - 1H, 13C and 15N assignment of the Isl-1 homeodomain. PMID- 9752007 TI - [HLA-Cw genotyping of serologically and non-serologically defined alleles using sequence-specific primers (PCR-SSP) in a Shanghai Han population]. AB - The new classical complement-mediated microlymphocytotoxicity test can detect a total of 10 different serologically defined antigen specificities (CWl-10) encoded by HLA-Cw locus. However, the blank amounts to 20%-50% for there are Cw antigens which can not be detected with antisera available. We adopted a PCR-SSP method to genotype a sample of 70 subjects collected from a Chinese Han population in Shanghai. We identified for the first time the following non serologically defined HLA-Cw alleles in the Chinese Han population: Cw*1201/1202,1203,1301,14,1601,1602,1701, and also the serologically defined HLA Cw alleles such as: Cw*01,02,0302/0304,0303,04,0501,0602,0701/0702/0703,08. With this sample we have made a survey of HLA-Cw allele frequencies in a Chinese Han population in Shanghai, with blank frequency being lowered from 0.307 to 0.028. The Cw*08 was originally considered to be rare in Chinese population. However, we found 12 cases of Cw*08 in this sample, which clearly does not coincide with the original point of view. Our results confirmed that PCR-SSP typing is rapid and accurate for HLA-Cw alleles. PMID- 9752008 TI - [Relationship between the occurrences of AL, MDS and AA and abnormal BM proliferation of patient's parents]. AB - After studying of familial leukemias, Myelodysplastic Syndrome (MDS) and aplastic anemia (AA), we observed and analysed bone marrow (BM) cells hematologically and molecular-cytogenetically in 36 persons who are first degree relatives (FDRs) of patients with acute leukemias (AL), MDS and AA. The peripheral blood (PB) lymphocyte chromosome fragility sensitive to folic acid and unstability was also analysed in 18 FDRs. The abnormal BM megakaryocystic/erythroid cellularity and the rearrangement of c-erbB were found in 66%-86.1% of parents and siblings of patients. The associations of dysplastic megakaryopoiesis, including the presence of lymphoid small megakaryocytes, with the chromosomal monosomy or/and the rearrangement/amplification of C-erbB, were found in a few parents and siblings. These results were consistent with those of MDS, Fanconi Anemia (FA) and AL. The normal karyotype and SCD positive of BM cells and PB lymphocytes, and PB lymphocyte chromosomal fragility and unstability were found in most of patients' parents, while familial chromosomal monosomy of BM cells and PB lymphocyte chromosomal fragility were found in parents and siblings of familial leukemia patients. Based on the studies of a large family with 7 cases of acute erythroleukamia and relative myeloleukemias in three consecutive generations and a family with 3 CAA and 1 AML, the rearrangement of c-erbB might be inherited. The rearrangement/amplification of c-erbB and its PCR detected results could be the indicators of gene diagnosis of preleukemia and might be useful in genetic conselling of leukemias. The common origin of AL, MDS and AA was discussed. PMID- 9752009 TI - [Distribution of E- and St-specific RAPD fragments in few genomes of triticeae]. AB - Two E genome--(including Ee and Eb) and two St-genome-specific RAPD markers were successfully cloned. Sequencing data indicated that the four DNA fragments were reported for the first time. Chromosome fluorescent in situ hybridization (FISH) showed that OPD-12(444) (E-specific) was a highly tandem repeat. OPF-03(1296) (Eb specific), OPB-08(525) (St-specific), and OPN-01(817) (St-specific) were highly dispersed repetitive sequences. The FISH and Southern hybridization of genomic DNA also revealed that all of the four highly repetitive sequences were shared among relatively closer genomes of triticeae. The difference was mainly in the fragment length and the copy number. Whether a genome can amplify a specific sequence via RAPD is mainly dependent on if there are primering sites at both ends of this sequence in the genome. We also discussed the potentiality of these highly repetitive DNA fragments in detection of alien chromatin in wheat and determination of genome constitution of polyploid triticeae species. PMID- 9752011 TI - [The basic trends in the development of health for the labor potential of Russia]. PMID- 9752010 TI - [Expression regulation of purine biosynthetic genes in Salmonella typhimurium. VI. Isolation and characterization of super-repressor mutants]. AB - Starting from strain of Salmonella typhimurium purD::lac, 86 exponential cultures were mutagenized with NTG and white or light blue clones on E + ado + Xgal plate were selected as candidates of purRs mutant. Total 66 independent candidate strains were obtained. By assaying their beta-galactosidase activity under the repressed and derepressed conditions, determining their frequency of revertional mutation, Conducting transductional analysis of mutational site and dorminance test, 11 candidates strains were proved to be super-repressor mutants. These mutants are useful for studying the expression of purine biosynthetic gene and relationship between protein structure and function in general. PMID- 9752012 TI - [The scientific trends and tasks in occupational physiology today]. AB - Evaluating a human during occupational activities, occupational physiology should put emphasis on two principal and cooperated directions. Those are aimed to minimize occupational risk, preservation of working capacity, long occupational life and human health. The first direction is associated with theoretic and practical work on differential diagnosis of various functional systems: regulation of working strain, fatigue and tension. Work in this direction helps to solve such practical problems as rehabilitation and assessment of human resources, evaluation of occupational stress, design of automated monitoring systems, etc. The second direction covers hygienic regulation of occupational factors. Their intensity and duration should be assessed with consideration of age, personality and other traits of a worker. In addition, combined action of existing hazards and the new ones should be considered in formation of regulatory technology documents. PMID- 9752013 TI - [The organization of automated information systems on occupational diseases and of the hygienic monitoring of working conditions]. AB - Creation of automated database "Archive of occupational diseases clinic" is an important task of informational support for occupational pathology centers. Constructing the database, the authors designed an "Accounting chart of patient with occupational disease for diagnostic survey in occupational pathology center" and a "Diagnosis of occupational disease" code book adjusted to X ICD with instruction. Analysis of the information obtained forms a basis for forecasting the course of pathologic process, for justifying a complex of treatment and prophylaxis. PMID- 9752014 TI - [From industrial hygiene for women to the protection of the reproductive health of workers. Principles and outlook]. AB - The issues and prospects of workers' reproductive health as a problem of occupational health in nowadays Russia are outlined with due account for WHO, ILO, EC, etc. concepts and recommendations. Main principles of reproductive health protection are gender approach, additional protection of vulnerable groups, e. g. pregnant women and obligatory account for combined effects of occupational, environmental and socio-economic risk factors. Two groups of criteria are essential: criteria of health and criteria of emancipation. Criteria for evaluation of reproductive health disorders are worked out based on official statistical indices and it is proposed to rate some of these as of extra category class for effective risk assessment and risk management. The strategy and priorities in workers' reproductive health are discussed and need for international cooperation is noted for resolving this problem. PMID- 9752015 TI - [The basic results of and outlook for scientific research on the problems of occupational medicine and industrial ecology in the central Volga region]. PMID- 9752016 TI - [The current problems of industrial medicine in the Kyrgyz Republic]. PMID- 9752017 TI - [The results of and outlook for scientific research on industrial medicine and industrial ecology in Georgia]. PMID- 9752018 TI - [The achievements in industrial hygiene and occupational pathology and the outlook for the development of industrial medicine in Azerbaijan]. PMID- 9752019 TI - [Industrial medicine in Latvia]. AB - The authors have reflected new trends in occupational health in Latvia. At present the whole system of general health care in Latvia is under consolidation and reconstruction. Thus, the occupational health and safety system of Latvia, as a nature part of general health care system is also under changes and reconstruction. Today Latvia's workforce consist of 1.68 million people, working in a large number of workplaces, mainly in industry and agriculture. There are still very hazardous working conditions in many workplaces. The total number of diagnosed occupational diseases ranges from 90 to 220 cases per year. If counted per 100,000 workers the rates of occupational diseases are 5.1 in 1981 and 23.4 in 1995. New system of occupational health is under development in Latvia. New legislation and regulations are published and extensive training for specialists provided. A new system of information is under construction as well as new hygienic standards. Main stress is paid to ensure that standards of EC are met. PMID- 9752020 TI - [The scientific achievements of the Kazan State Medical University in studying the current problems of industrial medicine]. PMID- 9752021 TI - [For how long should antipsychotic medication be continued after the first psychotic episode in schizophrenics?]. AB - Four patients, two women aged 24 and 62 years and two men aged 25 and 24, respectively, were admitted because of psychosis. A Dutch consensus paper advises treating patients with a first episode of schizophrenia or schizophreniform disorder with neuroleptics for two years as secondary prophylaxis. However, this advice should be tailored to the individual patient's characteristics. Thus, the first patient was given prophylactic medication for five years because she had many schizophrenic symptoms and a positive family history. In the second patient, the diagnosis was much less certain and, because of her advanced age, the risk of developing tardive dyskinesia was considerable. Prophylaxis was given for three months only. The third patient used drugs and did not really want to be treated. In the fourth patient the affective symptoms could not be interpreted for certain as part of a basic schizophrenic defect. In addition, he would be seriously handicapped professionally if he developed tardive dyskinesia. In his case, two years of secondary prophylaxis was advised. PMID- 9752022 TI - [Anthelmintics]. AB - The anthelminthics registered in the Netherlands include albendazole, mebendazole and praziquantel. Available but not registered are ivermectin and diethylcarbamazine. With these five drugs nearly all indigenous and imported helminth infections in the Netherlands can be treated effectively. Neurocysticercosis is difficult to treat with medication, surgery may be needed. PMID- 9752023 TI - [Inhaler therapy for adults with obstructive lung diseases: powder or aerosol?]. AB - When prescribing inhalation medication in the ambulant treatment of patients with asthma and chronic obstructive pulmonary disease (COPD), a choice can be made between dry powder inhaler (DPI) and pressurized metered dose inhalers (pMDI). The degree of deposition in the lower airways depends on the dose delivery via the inhaler and the mean diameter of the released particles (MMAD). With a DPI, dose delivery and MMAD depend on the inspiratory flow rate. With a pMDI dose delivery and MMAD do not depend on the inspiratory flow rate but on hand-mouth co ordination. The main determinants for the choice of a DPI or a pMDI are the degree of co-operation and co-ordination, and the inspiratory flow rate of the patient. PMID- 9752024 TI - [Quality assessment indicators of cardiac surgeons' work; the validity of mortality rates]. AB - OBJECTIVE: To determine the possibility of comparing the mortality rates of patients operated by different heart surgeons with each other. DESIGN: Retrospective cohort study. SETTING: Academic Medical Centre, Amsterdam, the Netherlands. PATIENTS AND METHODS: Clinical information, operation data and follow-up data on 783 patients who had undergone cardiac valve replacement, were collected from the clinical records. Aortic valve replacement had been performed in 446 patients (1979-1986) and mitral valve replacement in 337 patients (1980 1990). RESULTS: The one-year mortality rate was higher among patients operated on by heart surgeon A than among patients operated on by the other heart surgeons from the same team, viz. 16.4% and 9.5%, respectively, an absolute difference of 6.9%. The 95% confidence interval of the difference was 1.7-12.9. However, it was also found that the risk profiles of these patients of surgeon A differed from those of the other patients. After multivariate correction for this difference in risk profile, the difference in mortality was no longer statistically significant. CONCLUSION: The differences in mortality observed in our study could not be attributed to difference in quality of the heart surgeons, but were related with the risk profiles of the patients operated by one of them. Thorough analysis with correction for risks is necessary for the assessment of the quality of care, if the conclusions are not to be misleading. PMID- 9752025 TI - [Management of psoriasis in family practice is now in closer agreement with the guidelines of the Netherlands Society of Family Physicians]. AB - OBJECTIVE: To assess changes in incidence of psoriasis and to study changes in the management of psoriasis in general practice after the sending of guidelines on management of psoriasis to general practitioners (GPs) by the NHG. DESIGN: Secondary data analysis. SETTING: Netherlands Institute for Health Care Research (NIVEL), Utrecht, the Netherlands. METHOD: Data on the incidence of psoriasis and its management by general practitioners were collected from a file predating the publication of the NHG guideline 'Psoriasis' in 1994, namely the 'Dutch National Survey of General Practice' (NS; 1987/'88), and from a subsequent file, the 'Registration Network Groningen' (RNG; 1995). RESULTS: In the NS there were 106 new patients with psoriasis (incidence: 1.3/1000/year; 95% confidence interval (95% CI): 1.2-1.4) while in the RNG there were 24 (incidence: 1.2/1000/year; 95% CI: 0.7-1.9). In all, there were 466 psoriasis patients in the NS and 125 in the RNG. The number of referrals to dermatologists was halved in 1995 (7.2%) compared with 1987/'88 (14.4%; p < 0.05). The most frequently prescribed dermatologica in psoriasis was in 1995 corticosteroid group 3 (32.8%; in 1987/'88: 28.5%), but the rise was stronger in corticosteroid group 2 (29.6%; in 1987/'88: 16.0%; p < 0.001) and group 4 (16.0%; in 1987/'88: 8.8%; p < 0.05). CONCLUSION: The incidence of psoriasis in general practice had not changed between 1987/'88 en 1995. Referral pattern and prescription shifted towards the guidelines issued by the NHG. PMID- 9752026 TI - [The influence of an ECG on patient management in family practice]. AB - OBJECTIVE: To determine how often electrocardiography, as an addition to history taking and physical examination, leads to changes in non-emergency patient management in general practice. DESIGN: Prospective observational study. SETTING: 18 general practices in Amersfoort and its surroundings, the Netherlands. METHOD: From the end of September, 1996 until the beginning of February 1997, the general practitioners (GPs) filled out two questionnaires each time an ECG was recorded, one before and one after the recording. The patient management planned by the GP before and after the availability of the ECG results was then compared. RESULTS: A total of 119 sets of questionnaires was obtained from 119 patients. In 47 patients (40%; 95% confidence interval: 31-48) the GP's management changed after recording and interpretation of the electrocardiogram. In particular, the decision whether or not to refer patients to a cardiologist was frequently changed (26 patients, 22%). The GPs management changed more often in patients in whom palpitations and dyspnoea had been the reason for taking an ECG. CONCLUSION: The use of electrocardiography in general practice in non-emergency situations would seem a valuable instrument in addition to history taking and physical examination. PMID- 9752027 TI - [Three hypotonic neonates with hypertrophic cardiomyopathy: Pompe's disease]. AB - Three neonatal patients, one girl and two boys, presented with infantile Pompe's disease. A generalized hypotonia with decreased tendon reflexes and heart failure due to hypertrophic cardiomyopathy dominated the clinical picture in all three; these symptoms are uniformly and characteristically present. This autosomal recessive glycogen storage disease is caused by a deficiency of lysosomal alpha glucosidase. The diagnosis, suspected on the basis of the characteristic clinical picture and the results of simple laboratory tests, is made by measurement of the enzymatic activity or DNA analysis. Most patients die in their first year of life, no treatment being available. PMID- 9752028 TI - [Continuous pressure is preferred to flow triggering of respiration in the apnea test following the protocol for brain death determination]. AB - The apnea test is part of the brain death protocol of the National Health Council. If the patient is being given positive end-pressure respiration, he must not be uncoupled from the respirator. The apnea test should then be done by means of continuous positive airway pressure. Pressure triggering rather than the extremely sensitive flow triggering should then be chosen to trigger the respiration, since otherwise the patient may unjustifiably be declared 'not brain dead' as a result of slight aspecific movements (bumping against the bed, beating of the heart). PMID- 9752029 TI - [Feeding tubes for enteral nutrition]. PMID- 9752030 TI - [Feeding tubes for enteral nutrition]. PMID- 9752031 TI - [Feeding tubes for enteral nutrition]. PMID- 9752032 TI - [Therapeutic management of agitation in the dying; more than just sedation needed]. PMID- 9752033 TI - [Clenched fist injuries from teeth: not to be disregarded]. AB - During the delivery of a blow to the jaw, two men, 33 and 34 years of age, suffered an injury at the level of the right metacarpophalangeal joint in, respectively, the 4th and 3rd digit. In both cases, purulent arthritis and destruction of the MCP-joint developed. Clenched-fist injuries are known for their severe complications such as septic arthritis, osteomyelitis and persistent infection leading to amputation. These complications are due to the easy perforation of the MCP-joint capsule and the fact that the patients do not seek medical treatment until a significant inflammatory process has developed. Exploration of the wound on a flexed hand is crucial to exclude perforation of tendon, joint and bone. The wound should be left open to avoid infections. In case of infections, which can be caused by a variety of aerobic and anaerobic bacteria, the recommended treatment is immediate debridement and administration of broad-spectrum antibiotics. PMID- 9752034 TI - [Liver and artificial liver]. AB - Despite good results of orthotopic liver transplantation in patients with fulminant hepatic failure the need still exists for an effective and safe artificial liver, able to temporarily take over the complex liver function so as to bridge the gap with transplantation or regeneration. Attempts to develop non biological artificial livers have failed, mostly when controlled clinical trials were performed. In the last decade several different types of bioartificial livers have been devised, in which the biocomponent consists of freshly isolated porcine hepatocytes or a human hepatoblastoma cell line. The majority use semipermeable hollow fibers known from artificial kidney devices. The liver cells may lie either inside or outside the lumen of these fibers. In vitro analysis of liver function and animal experimental work showing that the bioartificial liver increases survival justify clinical application. Bioartificial livers are connected to patients extracorporeally by means of plasmapheresis circuit for periods of about 6 hours. In different trials about 40 patients with severe liver failure have been treated. No important adverse effects have not been reported in these phase I trials. Results of controlled studies are urgently needed. As long as no satisfactory immortalised human liver cell line with good function is available, porcine hepatocytes will remain the first choice, provided transmission of porcine pathogens to man is prevented. PMID- 9752035 TI - [Gastroesophageal reflux disease: pathophysiology, diagnosis and drug therapy]. AB - The principal mechanism leading to gastro-oesophageal reflux is an increased frequency of transient lower oesophageal sphincter relaxations; other factors are oesophageal hypersensitivity to gastric juice, hiatus hernia, and possible duodenal reflux. Patients with classical symptoms such as heartburn and regurgitation may be treated pharmaceutically combined with life style counselling. If the symptoms have not improved after 6 to 12 weeks, endoscopical examination is performed and, if necessary, 24-hour pH monitoring, barium radiographing and manometry. In the case of atypical symptoms such as dysphagia, laryngitis, asthma and chest pain, there is more reason to pursue diagnostic testing. In patients with dysphagia endoscopy is indicated to exclude malignancy. Drug treatment can be subdivided into antacids, H2 receptor antagonists, cytoprotective agents, prokinetics and proton pump inhibitors. In general practice a step-up approach to treatment is preferable, while for specialist treatment a stepdown approach is more (cost-)effective. Drawbacks of medical treatment are considerable frequency of recurrence of oesophagitis, persistence of regurgitation in 'volume refluxers' and controversial data on the possible development of (pre)malignant lesions of oesophagus and stomach. Surgical treatment is a good alternative for patients with persistent severe regurgitation during medical therapy and for young patients who prefer surgery to lifelong medication. Patients with Barrett's oesophagus should undergo regular endoscopic biopsy surveillance. PMID- 9752036 TI - [Surgical treatment of gastroesophageal reflux disease]. AB - Antireflux surgery is successful in 85-90% of eligible patients, with relief of symptoms, cure of oesophagitis and possibly prevention of progression of the dysplasia in a Barrett's oesophagus. The mortality in the latest publications is given as 0.05%. The morbidity, apart from recurrences, is not yet sufficiently known. Some 250 antireflux operations are performed annually in the Netherlands, fewer than 20% of the estimated requirement of 10 operations per 100,000 of the population per year, and also fewer than in Scandinavia. Nissen fundoplication (folding the fundus of the stomach around the entire circumference (360 degrees) of the oesophagus) is generally accepted as the standard primary operation. Nissen fundoplication during laparoscopy seems to be just as good. Results of randomized clinical trials will have to be awaited to prove this assumption. Belsey's operation (folding the fundus around 270 degrees of the circumference of the oesophagus via thoracotomy) is nowadays performed almost exclusively in recurrent reflux disease and in persistent dysphagia after a primary operation. PMID- 9752037 TI - [Good short-term and mid-term results after laparoscopic Nissen fundoplication for gastroesophageal reflux]. AB - OBJECTIVE: Analysis of per- and postoperative complications and functional results in the short and medium-long term following Nissen laparoscopic fundoplication for reflux oesophagitis. DESIGN: Prospective, descriptive. SETTING: Academic Hospital of the Free University in Amsterdam and Medical Centre Alkmaar, the Netherlands. METHOD: Between January 1992 and June 1997 in both hospitals, 50 patients with reflux oesophagitis were operated on laparoscopically and investigated pre- and postoperatively according to a protocol involving scoring of symptoms, endoscopy and pH-measurement; in some of the patients, manometry, gastric emptying studies and an insulin provocation test were also performed. RESULTS: There was no surgical mortality. Peroperative complications were seen in five patients. Conversion to laparotomic fundoplication was necessary in five patients. Five patients had mild postoperative complications. The mean hospital stay after operation was 5.5 days. Three to six months after discharge 90% of the patients reported the functional results to be good or perfect (n = 100). After a mean follow-up of 48 months this was 97% (n = 30); recurrence was seen in two patients (7%). Three patients had to be re-operated for severe dysphagia. CONCLUSION: Laparoscopic fundoplication performed by an experienced surgeon is a good technique offering definite advantages to the patient. The short and medium-long term results are good. PMID- 9752038 TI - [Good results of laparoscopic Thal fundoplication for treatment of gastroesophageal reflux in children]. AB - OBJECTIVE: Evaluation of the results of laparoscopic fundoplication according to Thal as a treatment for gastro-oesophageal reflux in children. DESIGN: Prospective, descriptive. SETTING: Department of paediatric surgery, Wilhelmina Children's Hospital, Utrecht, the Netherlands. METHODS: Between November 1993 and May 1996, 53 children with reflux and an average age of 6 years underwent laparoscopic fundoplication according to Thal; in 23 of them a gastrostomy was created as well. The most important symptoms were vomiting and lack of growth. There were 18 children with primary reflux and 35 with secondary reflux, 28 of whom had psychomotor retardation. Preoperatively all patients were subjected to 24-hour oesophageal pH-monitoring, upper GI series and oesophagogastroscopy. Pathological reflux was defined as a pH < 4 during 5% or more of the total time measured (24 hours). pH-monitoring was repeated three months postoperatively to evaluate the effect of the fundoplication and esophagogastroscopy was repeated in case of preoperative oesophagitis. RESULTS: In one patient the laparoscopic approach was converted peroperatively because of bleeding that had stopped on exploration. Enteral feeding could be recommenced on day one in 49 of 53 children. The mean hospitalisation time was 4.4 days. One patient was reoperated for a too tight fundoplication and two patients died of unrelated causes. The mean follow-up period for the other 50 patients was 11 months (1-35). On follow up, 42 of them (84%) were free of symptoms, while 31 of 41 children (76%) in whom pH-monitoring could be done 3 months postoperatively displayed no recurrence of pathological reflux. CONCLUSION: Thal fundoplication can be performed laparoscopically in children. Enteral feeding becomes possible again quickly and the period of hospitalisation is short. PMID- 9752039 TI - [Femoral shaft fracture in children younger than 4 years: shorter hospital stays with the help of at home traction apparatus]. AB - OBJECTIVE: Evaluation of home traction as a treatment as a treatment of femoral shaft fractures in children with the objective to shorten the hospital stay. DESIGN: Retrospective. SETTING: Paediatric Surgical Centre Amsterdam (EKZ/AMC and AZVU), the Netherlands. METHOD: In the period 1991-1995, 18 femoral shaft fractures in children younger then 4 years were treated. In ten of them traction was applied at home (in the other cases the parents refused to co-operate, the home situation was not appropriate, there were additional medical problems or there was a suspicion of child abuse). During follow-up of the group treated at home with traction, angulation, deformity and leg length discrepancy were determined with special attention to complications. The parents' experience of this method was evaluated by telephone (n = 8). RESULTS: The median age of the children was 2.4 years. The mean hospital stay was 7 days (range: 3-12), the mean follow-up 2.4 years (range: 1.0-4.3). Angulation, rotational deformities and leg length discrepancy > 1 cm did not occur. Oedema and pain were seen in 1 patient as a result of incorrect treatment at home. In one patient a compartment syndrome occurred after a switch from traction to a plaster treatment in another hospital. With exception of some small practical and informational problems, parents were very pleased with this method. CONCLUSION: Treatment at home of femoral shaft fractures in children with traction is a simple and effective method which reduces the hospital stay to one week with minimal complications. Good patient selection and instructions of the parents are mandatory. PMID- 9752040 TI - [Inguinal pain caused by iliopsoas bursitis]. AB - Four patients, three men aged 69, 55 and 45 years and one woman aged 72 years, complained of pain in and around the inguinal region without clear cause. The crippling symptoms were of a few days' to 8 years' standing. Passive hip movements were not restricted. Presence of pain after pressure on or stretching of the bursal wall during provocation tests and absence of other disorders resulted in a probability diagnosis of iliopectineal bursitis. This diagnosis was confirmed by marked abatement of the pain after an injection of lidocaine into the bursa. Inflammation of the iliopectineal bursa may occur after injury or overstrain and cause various kinds of complaints. The diagnosis injection of lidocaine as a rule constitutes an adequate treatment as well. PMID- 9752041 TI - [Cholestatic hepatitis ascribed to the use of thiabendazole]. AB - Two women, aged 27 and 42 years, both born in Surinam and both suffering from heterozygous thalassemia, developed cholestatic hepatitis three and two weeks respectively after the start of a two-day course of thiabendazol for Strongyloides stercoralis infection. Other causes of cholestasis were unlikely in view of the results of blood tests, echography and the endoscopic retrograde cholangiopancreaticography. The symptoms persisted for several months, and the liver function disorders for 7 years and one year, respectively. The incidence of thiabendazole-induced cholestatic hepatitis is unknown, but probably low. PMID- 9752042 TI - ['Public health status and perspectives' 1997. VI. Effects of health care]. AB - The aim of this study was to investigate whether more health improvement might be attained with existing medical interventions. The interventions for ten diseases were studied to assess their efficacy (effect in ideal, experimental situation), their effectiveness (effect in daily practice) and the discrepancy between the efficacy and effectiveness. In addition, the causes of unexpectedly low effectiveness were studied. Interventions with proven efficacy were identified for all ten diseases. The effectiveness was less often studied. Existing health care registrations could provide information on the effectiveness for only three diseases (tuberculosis, colorectal tumours, and acute myocardial infarction). Causes of unexpectedly low effectiveness were observed in several of the five phases of the process of care: contacting the health care system, diagnosing the disease, prescribing the appropriate intervention, executing the intervention and patient compliance. PMID- 9752043 TI - ['Public health status and perspectives' 1997. VII. Health care needs and health care consumption]. AB - In the 'Public health status and forecasts' 1997 attention is given to the relationship between health status and health care. The theme report 'Health care need and health care consumption' integrates information about both phenomena and about waiting lists in the different sectors of health care. It is difficult to quantify the need for health care, because statements about need always imply a judgment by parties involved. In the literature need for health care is often operationalized by historical data on health care consumption or by health status indicators. At the national level only limited quantitative information is available to support policy on waiting lists and waiting times. The data are seldom disease-specific. Changes in size and distribution (by age and sex) of the population will increase health care cost over the period 1994-2015 in the Netherlands by 0.9-1.0% per year. More detailed demographic projections, however, indicate that there are large disease-specific differences. PMID- 9752044 TI - [No additional benefit derived from determination of serum lactase levels for the evaluation of a patient with an acute abdomen]. PMID- 9752045 TI - [Pharmacotherapeutic compass 1998]. PMID- 9752046 TI - [Iatrogenous pressure pneumothorax due to incorrect placement of the feeding tube]. PMID- 9752047 TI - [Detection of metastatic melanoma in sentinel lymph nodes by means of polymerase chain reaction]. PMID- 9752048 TI - [Higher hopitalization rates for schizophrenic Surinamese in Netherlands cannot be attributed to theimmigration of higher numbers of preschizophrenic patients]. PMID- 9752049 TI - [Right-sided diverticulitis masquerading as acute appendicitis]. PMID- 9752050 TI - [Right-sided diverticulitis masquerading as acute appendicitis]. PMID- 9752051 TI - [Pain in the hip area accompanied by a fever]. AB - In 3 patients, 2 women aged 16 and 64 years and 1 man aged 64 years, with pain in the left hip region and fever, the diagnosis psoas abscess was made. After antibiotic treatment and drainage they recovered well. The primary from of psoas abscess is presumably caused by haematogenous spread of bacteria, mostly Staphylococcus aureus. The secondary form is caused by spread of infection from surrounding tissue, mostly gastrointestinal micro-organisms with Crohn's disease and diverticulitis. Painful passive extension and endorotation as well as a painful flexion stress-test of the hip joint can indicate a psoas abscess. Echography and blood cultures should be performed if a psoas abscess is suspected. If echography is inconclusive, CT-scan can establish the diagnosis. The psoas abscess should be treated by percutaneous or surgical drainage combined with antibiotic therapy. The underlying cause of a secondary psoas abscess should be treated separately. PMID- 9752052 TI - [Public health in a new coalition agreement]. AB - In drawing up a new coalition agreement in the Netherlands, public health and health care will be important issues. The government now has the opportunity, by planning well, to correct the negative image of public health care (expensive and inefficient) and to cope with major problems, such as long waiting lists and unequal access to care. Requirement are agreement on a realistic percentage of volume growth, based on demographic trends, good agreement on conditions of employment efficient use of funds which citizens can understand, limiting drug prices and outlining a health care system for the future that is transparent and universally accessible, with collective responsibilities clearly delineated. Furthermore research on public health and health care should be stimulated. PMID- 9752053 TI - [Prognosis of the total hip prosthesis]. AB - The prognosis of total hip arthroplasty is excellent and as many publications show, the survival of a good type of implant is well above 90 per cent at the 10 year follow-up. The results of the average orthopaedic surgeon may not match those obtained by the experts who have published these results. Therefore, in 1979 a prospective multicentre study was started in Sweden to evaluate the outcome of total hip arthroplasty. This National Hip Register has shown that the outcome is related to type of prosthesis, patient selection and operative technique. The results from one orthopaedic department to another differed considerably. The feedback from the Register resulted in improvements of the overall outcome and decrease of the differences between the orthopaedic departments in Sweden A Dutch National Implant Register is needed. PMID- 9752054 TI - [Total hip replacement arthroplasty in the Netherlands]. AB - The total hip arthroplasty (THA) is an effective treatment of osteoarthritis of the hip joint. Each year, more than 16,000 THAs are performed in the Netherlands. The incidence of THA had doubled between 1980 and 1994. The medical indication has extended to younger and older patients. Due to the success of the THA the number of alternative treatments, such as arthrodesis and osteotomy decreased. The number of revision surgeries increased more than four folds, to 1,400 in 1994. Based on these data the increase of primary and revision hip arthroplasties is expected to continue, but it is unknown how much the exact increase will be. In view of the increased demand it is important to gain more insight into the numbers of primary and revision surgical procedures to be expected, so that a better estimation can be made of the health care capacity and financial resources required in the future. PMID- 9752055 TI - [Surgical techniques used in the revision of hip prostheses]. AB - The most common cause of failure of cemented and cementless total hip arthroplasties is aseptic loosening, a slow but progressive process that often results in loss of bone stock. The diagnosis of loosening of the prosthetic component is difficult and depends mainly on migration and change in position of the prosthesis and the appearance of clear zones around the prosthesis on X-ray photographs. The key problem in revision surgery is how to manage the periprosthetic bone loss. Controversy exists about the best treatment for bone stock defects. The various techniques are directed at restoration of the hip mechanics, restoration of the defects in the osseous wall, replacement of the resorbed bone and thus restoration of the functional stability of the joint. PMID- 9752056 TI - [The relationship between genetic polymorphisms and disease, illustrated by the renin-angiotensin-aldosterone system and cardiovascular disease]. AB - The role of molecular genetics in the pathophysiology of various diseases is becoming clearer and clearer. In the field of cardiovascular diseases, molecular genetic aspects have been shown to play a definite role in the aetiology of these diseases. Several molecular-genetic variations called polymorphisms, occur in the population. The genes encoding the different components of the reninangiotensin aldosterone system (RAAS), an important system in the regulation of the function and structure of the heart and vascular wall, also display polymorphisms. For some of these polymorphisms associations with various cardiovascular and renal diseases have been described. At present, this is particularly clear for the relation between angiotensin-converting-enzyme (ACE) polymorphism and the incidence of atherosclerotic complications and diabetic nephropathy, and for the relation between so-called M235 T-variant of the angiotensinogen gene and hypertension. Future research will have to show where it is worthwhile to use these and other polymorphisms as a marker for genetic risk. In what way the different RAAS-polymorphisms relate to functional abnormalities is as yet unclear, as are the potential therapeutic implications. PMID- 9752058 TI - [Good results 5 years after surgery for proximal femur fractures]. AB - OBJECTIVE: Determining the results 5 years after surgery for proximal femur fractures. DESIGN: Descriptive, retrospective. SETTING: Department of General Medicine, Red Cross Hospital, the Hague, the Netherlands. METHOD: In 1996, for all patients operated on for proximal femur fractures in 1991, data were collected from the patient records regarding the admission and situation at home, and from the municipal archives, the family physician, the patient or his or her family regarding survival. The 5-year survival was compared with that of a cohort matched for age and sex, according to data from the Central Bureau for Statistics. The level of function after 5 years was evaluated by means of a Broos checklist. RESULTS: Of the 117 patients operated, 69 had medical, 9 lateral and 39 pertrochanteric fractures. The average age of the 20 men and 97 women was 71 and 82 years, respectively. The average duration of hospitalisation was 31 days. Seven patients (6%) died while still in hospital. Of the 110 surviving patients, 61 (55%) returned to the situation in which they lived before. Starting 6 months postoperatively, the 5-year survival curves were parallel to those of the matched cohort. After 5 years, 53 patients (45%) were still alive. Of these, 37 patients (70%) functioned well. CONCLUSION: Most patients with proximal femur fracture belong to the category of patients in advanced old age. The survival after 5 years was 45%; most of the mortality occurred during the first 6 months after the operation. Of those surviving, 70% functioned well. PMID- 9752057 TI - [An estimate of the intramural costs of the placement of a total hip prosthesis]. AB - OBJECTIVE: Determining the intramural costs of the (re) placement of a total hip prostheses. DESIGN: Retrospective, descriptive. SETTING: Utrecht University Hospital (AZU) and the De Weezenlanden Hospital in Zwolle (ZWZ), the Netherlands. PATIENTS AND METHOD: From the records of 137 patients in whom an uncomplicated primary placement or revision of 151 total hip prostheses had taken place in the AZU or the ZWZ during the period 1990-93, data were collected on the number of days hospitalised, the length of the operation, the medical materials used, the number of radiological procedures, ECG's and laboratory determinations, and the visits to the outpatient clinic. The costs of a hip replacement were determined on the basis of the salary costs, tariffs and purchase prices. RESULTS: The average total cost was f 9381 for the placement of a cemented prosthesis, f 10,796 for the placement of a cement-free prosthesis, and f 17,879 for a revision operation. About 90% of the total costs were due to the price of hospitalization, surgery and prosthesis. With 14,000 primary placements and 1400 revisions annually, the cost is approximately 186 million guilders. An expected increase in the number of hip replacements and their cost should be taken into consideration. PMID- 9752059 TI - [Revised consensus 'Diagnosis of the dementia syndrome']. AB - Dementia is a clinical syndrome and is diagnosed on clinical grounds. Various types can be distinguished: the Alzheimer-type, frontal lobe dementia and subcortical dementia syndromes. Neuropsychological examination can contribute to the clinical diagnosis. Differentiation from delirium and depression, which may co-exist with dementia, is necessary. Once a dementia syndrome has been diagnosed its cause has to be ascertained. Alzheimer's disease is the most common cause and can often be diagnosed clinically. The clinical suspicion of vascular dementia has to be confirmed by imaging methods. Drug intoxication may cause or contribute to dementia. Blood tests should be performed routinely, but EEG, CT or MRI, SPECT and genetic tests can be carried out on clinical indication. Subsequently the need for care of the patient has to be established, as well as the ability of the carers to meet it. Regular follow-up is necessary. A definite diagnosis can only be made post-mortem when neuropathological examination has been performed. The organisation of diagnosis in the dementia syndrome should preferably take place in specialised multidisciplinary teams. PMID- 9752061 TI - [Presurgical diagnostic stereotaxic biopsy is a good predictor of breast cancer in patients with suspicious deviations on mammograms]. PMID- 9752060 TI - [Recombinant factor VIIa (eptacog alfa): a new therapeutic option for patients with severe bleeding disorder due to inhibitory antibodies against a coagulation factor]. AB - In 2 patients with severe haemorrhage (a 63-year-old man with haemophilia A (the factor VIII level was 29%) and a 44-year-old woman), of an inhibitory antibody against factor VIII was diagnosed. The development of recombinant factor VIIa (eptacog alpha) has made available a new therapeutic option for patients with an inhibitory antibody against a coagulation factor. Both patients were treated successfully with the new factor after other forms of treatment had failed. The new concept of the coagulation cascade on which the treatment with eptacog alpha is based assumes that the lack of an amplifying loop in the coagulation which takes place via factor IX (in combination with factor VIII) can be compensated by extra stimulation of the principal route (tissue factor-factor VIIa --> factor X) by pharmacological amounts of factor VIIa. PMID- 9752062 TI - [The recommended daily amount of folic acid is insufficient for optimum homocysteine levels]. PMID- 9752063 TI - [The recommended daily intake of folic acid is insufficient for optimum homocysteine level]. PMID- 9752065 TI - [The bikini incision: nice, but not without painful complications]. AB - The low transverse abdominal incision, described by Pfannenstiel, is, mostly because of its decent scar, the incision of choice for most gynaecological operations. Making this incision, the ilioinguinal nerve and the iliohypogastric nerve can be involved. In many cases this causes a lasting numbness in the region around the scar. Some patients have a lasting, radiating, invalidating pain, which can be relieved surgically. PMID- 9752067 TI - [Inhalation therapy in children younger than 2 years. II. The practice]. AB - Given its pros and cons the indication for nebulisation therapy is limited. Nebulisation is cumbersome, expensive, time-consuming and often unnecessary even during severe bronchial obstruction. Inhalation is simple with metered dose inhalers and small inhalation chambers with low or no static charge and a mask over mouth and nose. Inhalation therapy in young children can fail on many points, with the risk that treatment is considered ineffective. Good instruction and control of correct use are mandatory. Inhalation therapy for small children has to focus on effective drug delivery, particularly during conditions like dyspnoea, tachypnoea and bronchial obstruction, because otherwise the therapy will fail when most needed. Of the three inhalation chambers available for small children, viz. the Babyhaler, the Aerochamber and the metal Nebuhaler, the last two are to be preferred. Since in the near future hydrofluoroalkane (HFA) aerosols will replace chlorofluorocarbon (CFC) aerosols an increased bronchial deposition has to be taken into account. PMID- 9752066 TI - [Inhalation therapy in children younger than two years. I. From theory to practice]. AB - Effective inhalation of drugs, even by small children under 2 years, is often faster, simpler, cheaper and better with metered dose inhalers with small antistatic (metal) inhalation chambers than with nebulisation. This is also true during considerable bronchial obstruction. It is mandatory that the inhalation chamber has a small dead space and well functioning valves opening at low flows. Effective dosing in small children is enhanced by more doses, given separately, while choosing the highest dose per spray available. Important factors determining bronchial deposition in small children are breathing frequency, tidal volume and the degree of bronchial obstruction and nasal obstruction, since inhalation goes primarily through the nose. If well-performed medication with a small inhalation chamber is clinically ineffective, it is better to start systemic medication, e.g. a corticosteroid, or even to consider artificial ventilation, rather than to try nebulisation. Better effective deposition is possible with inhalation of drugs in hydrofluoroalkane (HFA) aerosols, which will replace chlorofluorocarbon (CFC) aerosols in the near future. PMID- 9752069 TI - [Asthma in young children: a different approach to treatment by the family physician vs the pediatrician, a follow-up of symptoms over a period of 1 to 2.5 years]. AB - OBJECTIVE: To compare the therapeutic approaches to asthmatic symptoms in young children of general practitioners and paediatric pulmonologists, and to investigate the outcome of asthma in young children. DESIGN: Retrospective, descriptive. SETTING: Paediatric pulmonology outpatient clinic, University Hospital, Groningen, the Netherlands. METHOD: Charts of all 91 children younger than 2 years of age who were newly referred for recurrent cough and wheeze (asthma) between January 1, 1994, and September 30, 1995, were reviewed. Data were collected on clinical characteristics, drugs prescribed by the general practitioners, results of laboratory tests, and treatment prescribed by the paediatric pulmonologists. All children were followed up for periods of 12 to 30 months. RESULTS: Sixty-one children (67%) had been treated with antibiotics or with oral anti-asthma drugs by their general practitioners. No child had a positive radioallergosorbent test for inhaled allergens. Paediatric pulmonologists most commonly prescribed inhaled corticosteroids and bronchodilators to these patients. After 12-30 months of follow-up, 48 patients (53%) had no further symptoms. The only factor statistically significantly related to persistence of asthmatic symptoms during follow-up was aggravation of complaints by weather influences (odds ratio: 4; 95% confidence interval: 1-12). CONCLUSION: The common practice of prescribing antibiotics and oral anti-asthma drugs to young children in general practice is contrary to recent consensus reports on the treatment of asthma. Even in young children referred to specialists with asthmatic symptoms, such symptoms are commonly transient. Aggravation of symptoms by weather influences, which may be an expression of bronchial hyperresponsiveness, is a risk factor for persistence of symptoms. PMID- 9752068 TI - [Clinical decision making in family practice. A patient with abdominal pain and chills]. AB - An 82-year-old man was admitted because of abdominal pain and a shaking chill. His medical history revealed ileocecal resection because of ileitis associated with a Yersinia infection 3 years before admission. One year later he was readmitted because of bowel obstruction due to recurrent ileitis. He was treated with trimethoprim-sulfamethoxazole for two weeks because of positive serological tests for Yersinia and made a full recovery except for chronic diarrhoea. On the current admission, stool cultures yielded Campylobacter upsaliensis. Further analysis showed severe non-specific ulcerative ileitis without colitis. A diagnosis of Crohn's disease was made. The patient was treated with prednisone and mesalazine and made a full recovery. The chronic diarrhoea disappeared. The course was complicated by a cerebro-vascular thrombosis and severe thrombocytosis due to polycythaemia vera. Treatment with hydroxyurea was effective in lowering the thrombocyte count. PMID- 9752070 TI - [Initial experiences with shorter hospital stays after primary surgery for breast carcinoma]. AB - OBJECTIVE: To describe some personal, medical and financial consequences of moving up the discharge of patients from hospital after an operation because of breast carcinoma. DESIGN: Descriptive. SETTING: Department of Oncological Surgery, Medical Centre, Leiden University, Leiden, the Netherlands. METHOD: Thirty-five patients with breast cancer were operated during the period March to August 1997. Thirteen patients of this group were discharged sooner after operation, with the drain still in situ; the other 22 remained in hospital until after removal of the drain. Medical and financial consequences were investigated. RESULTS: The patient characteristics of the two groups were similar. In the group discharged earlier, the number of postoperative days in hospital on average was 4.5 days smaller. The number of postoperative complications in the two groups were similar; development of seroma after removal of the drain occurred less frequently in the group discharged earlier. The financial savings amounted to an average of Dfl. 2497.-per patient. The patients discharged earlier were very satisfied. CONCLUSION: The orientative study suggests that moving up discharge after a breast cancer operation is a policy that is safe, financially advantageous and satisfactory to the patients. PMID- 9752071 TI - [Severe tardive dyskinesia during treatment with risperidone and fluoxetine]. AB - A 26-year-old man with schizophrenia, disorganised type, and depression, developed severe tardive dyskinesia during treatment with risperidone and fluoxetine. In view of the course of the symptoms and the findings in the neurological analysis a causal relation with the use of the these second generation psychopharmaca was probable. These new-generation psychopharmaca are supposed to have fewer adverse events. Nevertheless, as is illustrated in this case, prescription is not without risk. Especially when using a combination of psychopharmaca, side effects must be monitored carefully. PMID- 9752072 TI - [Safeguarding the quality of health care in relation to cost effectiveness analysis]. AB - A meeting organized by Nederlands Tijdschrift voor Geneeskunde discussed the various interfaces of the quality of care, cost-effectiveness and health care. Cost-effectiveness analyses can contribute greatly to the quality of health care, but such research is methodologically complicated. Also, the effectiveness of particular interventions or therapies should be taken into account, if possible evidence-based, in the introduction of standards or guidelines for health-care workers. In practice, however, owing to various circumstances, the implementation of such guidelines leaves something to be desired. PMID- 9752073 TI - [Gentamicin dosing regimen for neonates: once daily]. PMID- 9752074 TI - [Gentamicin dosing regimen for neonates: once daily]. PMID- 9752075 TI - [Gentamicin dosing regimen for neonates: once daily]. PMID- 9752076 TI - [Gentamicin dosing regimen for neonates: once daily]. PMID- 9752077 TI - [Bone transplantation and bone replacement materials]. PMID- 9752078 TI - [Lead in the drinking water: a risk for bottle-fed infants; a report from the Health Advisory Commission]. PMID- 9752079 TI - [The history of the Yekaterinburg tragedy: the sources of the investigation into the death of the Romanov family]. AB - Presents main data on the life of the Romanov family after abdication of Emperor Nicholas II, about execution of family members and subjects in their attendance in Ipat'ev's house in Ekaterinburg, and burial of corpses. Offers a brief report on the circumstance of death of the family made by investigators A. P. Nametkin, I. A. Sergeev, and N. A. Sokolov in 1918-1924. Describes the activities of a group of enthusiasts headed by A. N. Avdonin and G. T. Riabov who searched for and found the remains of the imperial family and subjects in their attendance near Ekaterinburg. Presents the grounds for bringing an action on the basis of finding bone remains. PMID- 9752080 TI - [The participation of Yekaterinburg forensic medical experts in the expert assessment of the remains from a burial site near the village of Koptiaki]. AB - Enumerates grounds for experts evaluation carried out by Bureau of Forensic Medical Expert Evaluation of the Ministry of Health of Russia and Sverdlovsk Regional Bureau of Forensic Medical Expert Evaluation, lists the expert team members, and enumerates the methods of preparing the objects of expert evaluation (bone remains). Presents the results of assessing the date of burial, enumerates signs of exposure to thermal and chemical factors on the bones, and notes a callus on one skull. PMID- 9752081 TI - [The forensic medical assessment of the bone damages to skeletons from a group burial in the environs of Yekaterinburg]. AB - Results of comprehensive expert evaluation of bone remains found at the place of burial made 80 years ago are presented. Injuries inflicted by gunshots, mechanical, chemical, and other factors are classified. Diagnostic criteria for reconstruction of circumstances of death and burial of the Romanov family and subjects in their attendance are defined. PMID- 9752082 TI - [The detection of the traces of old damages to the bones of the cranial vault of the Emperor Nikolai II]. AB - Relevant reports are reviewed, primary medical files analyzed, and the skull of Nicholas II is investigated using roentgenography and computer-aided tomography with the aim of forensic medical diagnosis of traces of an old injury to skull vault bones. The Emperor had a superficial fracture of the skull vault bones. No periosteal callus is formed in the course of healing of such an injury, and the trace of the fracture is not seen beyond the contour of compact plate of the skull vault. The external compact plate at the site of injury was lost after long stay of the skull is soil at the place of burial, and therefore traces of a former injury could not be detected. PMID- 9752084 TI - [Determining if dismembered human remains belong to one or several corpses]. PMID- 9752083 TI - [The expert identification of the remains of the imperial family by means of molecular genetic verification of genealogical relations]. AB - The paper presents the results of international complex molecular genetic expert identification of skeletal remains of 9 subjects buried near the Koptiaki road in Ekaterinburg region, presumably belonging to the Romanov Royal Family and persons in their attendance. The armory of methods based on analysis of the least permissible amounts of DNA by the polymerase chain reaction and direct fluorescent sequencing of amplified DNA fragments included the latest scientific technologies. In addition, new methods were developed and used, which have no analogs in the world expert practice. The strategy included identification of biological gender of skeletons, of familial group among exhumed individuals, and of ties of relationship of this family with two independent maternal branches of the Romanov genealogical tree using tracing kindred markers based on mitochondrial DNA (mtDNA). The study was carried out in two stages: in 1992-1993 at Aldermaston Criminology Center of Home Office of the UK with participation of British specialists and in 1995 at Military Medical Institute of Ministry of Defence of the USA in Washington with participation of American specialists. In 1993 five skeletons were identified as Emperor Nicholas II, Empress Alexandra Fedorovna, and their three daughters with 99.6% certainty. From modern criminological viewpoint, the result could not be considered as sufficiently certain for such an extraordinary case, and therefore in 1995 molecular genetic studies of presumable remains of Nicholas II and his brother Prince Georgii Romanov were carried out again. The results showed absolute positional identity of mtDNA genetic code of these two men due to an extremely rare genetic abnormality (heteroplasmia), and thus the problem of appurtenance of the remains to the Romanov royal family was solved. PMID- 9752085 TI - [Regulations on the production of forensic medical expertise in the medical criminology laboratory departments of the Bureau of Forensic Medical Expertise. Appendix 4 to the order of the Ministry of Public Health of the Russian Federation of 10 December 1996 No. 407. Coordinated with the General Public Prosecutor, the Supreme Court and the Ministry of Internal Affairs of the Russian Federation]. PMID- 9752086 TI - [The Saint-Petersburg Ear, Nose, Throat and Speech Research Institute (a brief historical essay)]. PMID- 9752087 TI - [The characteristics of hearing perception in light of the problem of rehabilitating patients who have undergone implantation]. AB - There are significant differences in hearing perception of normal and cochlear implanted individuals. Though the implant processor changes time and frequency characteristics of the speech signal, some implanted patients are able to adapt to their new perception and to return to full understanding of live speech. We make the attempt to understand how the implanted patients percept new picture of speech and suggest the plan of rehabilitation for subjects who lost hearing early. PMID- 9752088 TI - [Treatment results in patients with juvenile angiofibroma of the nasopharynx]. AB - Surgical and radiation treatment outcomes are available for 46 patients with juvenile nasopharyngeal angiofibroma (JNA). Recurrences occurred in 17 (37%) of 44 operated patients. They were subjected to reoperation or radiotherapy. The latter is indicated in JNA recurrences, large-size tumors which are hard to approach, cranial cavity invasion. Immediate results of the treatment are favorable. PMID- 9752089 TI - [An exogenous psychotraumatizing factor as one of the causes for the development of neurotic reactions in subjects suffering from sensorineural hypoacusis]. AB - We performed special psychological investigations in 120 patients with neurosensory hypoacusis (NSHA) and emotional somatovegetative disorders. These patients were found to have neurosis and neurosis-like conditions. To elucidate the involvement of the exogenic factor on the onset of neurotic reactions in NSHA, we performed a questionnaire survey reflecting the attitude of the family, coworkers, strangers to people with hearing problems. It was found that neuroses and neurosis-like conditions in NSHA patients are provoked not only by premorbid personality traits but also by exogenic psychotraumatic factors. PMID- 9752090 TI - [Ways of using digital signal processing for expanding the potentials of hearing aids]. AB - Compared to analog hearing aids (HA), digital HA provide not only signal transformation but also adaptive filtration and reconstruction of the signals. This allows better speech perception in noisy surroundings for patients with neurosensory hypoacusis and recovery of speech perception in persons with partial frequency deafness. The design of digital hearing aids requires utilization of effective signal processors with signal memory. Use of the existing signal processors for such purposes enables construction of hearing aids with digital signal processing having suitable technical and maintenance characteristics. PMID- 9752091 TI - [Neuropeptides in the treatment of sensorineural hypoacusis in children]. AB - The examination of 250 children with neurosensory hypoacusis (NSH) demonstrated a 78% efficacy of endaural phonophoresis with dalargin (DPP) and transcranial electrostimulation (TCE) for improvement of acoustic function in the presence of positive ultrasound prognosis. Rheoencephalography and biochemical tests show that neuropeptides improve cerebral hemodynamics and normalize metabolism. Both DPP and TCE promote normalization of protein spectrum of the serum. In addition, TCE normalizes serum magnesium levels and its renal elimination, while DPP normalizes serum content of cholesterol. PMID- 9752092 TI - [The effect of Lasix on nystagmus in the caloric and optokinetic tests in Meniere's disease]. AB - The study was made of labyrinthine changes induced by lasix dehydration. Changes indicative of Meniere's disease were identified at comparison of the findings obtained by audiological lasix test, caloric optokinetic reactions. Four types of the nystagmic response in the bithermal lasix-test were determined. These reactions are helpful for more precise staging of the disease. PMID- 9752093 TI - [The phenomenon of the fixation suppression of caloric nystagmus in the diagnosis of vestibular disorders]. AB - Fixation nystagmus suppression in caloric tests was used in examination of 56 patients suffering from chronic remittent labyrinthopathy, acute labyrinthopathy, vestibular neuronitis, otosclerosis, adhesive otitis media, vertebrobasilar insufficiency, motor disease, chronic remittent vestibulopathy and neurinoma of the VIII nerve. The bithermal test (BT) with electronystagmography was made to calculate the fixation suppression index (FSI). Complete suppression of the caloric nystagmus in at least one BT test appeared to be the most frequent variant of the response (24 cases). This was not dependent on the severity of the vestibular analyzer lesion. In 3 cases FSI of the caloric nystagmus was rather high (> 0.50). Introduction of the period of glance fixation in performance of all the 4 BT tests may serve an additional method in differential diagnosis of the vestibular disorders. The fixation test results should be compared to findings obtained in the other vestibulometric tests. PMID- 9752094 TI - [The rehabilitation of children with the initial forms of conductive hypoacusis due to recurrent acute otitis media]. AB - The aim of the trial was study and rehabilitation of sound conduction functional disorders in 3-10-year-olds after recurrent acute otitis media. Tympanometry findings allowed to differentiate various alterations in the auditory tube and scarring in the middle ear. Treatment of the children was decided depending on the process in the middle ear. It is concluded that early diagnosis of adhesive phenomena is achieved with active search for latent auditory and tubular dysfunctions in children who previously had recurrences of acute otitis media. Children with sound conduction system dysfunction are recommended to be on the follow-up with control study of the auditory and tubular function once in 6 months and have repeat courses of corrective therapy up to stable effect. PMID- 9752095 TI - [The neurodystrophic component of the structural changes in the hemispheric cerebellar cortex of patients with acoustic neurinomas]. AB - Cerebellar specimens removed in surgical treatment of acoustic nerve neurinomas were studied morphologically in 16 patients. Histochemically, morphofunctional state of the vegetative adrenergic nervous fibers and endings of the vessels and connective tissue basis of the pia mater encephali in the region of the cerebellum indicates inhibited activity of the neuromediator contour of circulation and metabolism regulation in the brain. Light and electron microscopy in the cerebellar cortex detected disorders of circulation, nervous cell dystrophy, glyocytic proliferation. Correlation was found between activity of sympathetic component of the autonomic nervous system, severity of neurodystrophic changes in the cerebellar cortex and development of pathophysiological reactions arising intraoperatively and early after operation. PMID- 9752096 TI - [Experimental research on the ultrastructural changes in the cells of the spiral organ]. AB - Adverse effects of vibration and kanamycin on the spiral organ (SO), SO response to physiotherapeutic electrostimulation and ceruloplasmin were studied in guinea pigs. Vibration and kanamycin provoked the same reaction of the SO receptor cells which can be characterized as metabolic stress. In this case mitochondria suffer most of all. Electrostimulation and administration of protein-antioxidant ceruloplasmin gave rise to protective changes suggesting activation of biosynthetic and energetic processes in the receptor cells. PMID- 9752097 TI - [A comparative study of conservative treatment schemes in chronic tonsillitis in children]. AB - Three schemes of conservative treatment of chronic tonsillitis in children are compared. Chelidonium majus L. tincture improved tonsillar function, cellular and humoral immunity, nonspecific resistance, promoted a reduction in the number of recurrences. PMID- 9752099 TI - [An assessment of the immune status of the mucosal membranes in chronic rhinosinusitis]. AB - The authors studied immune status of upper respiratory tracts mucosa in patients with different forms of chronic rhinosinusitis (purulent, polypous, vasomotor) in comparison to control subjects. An original complex of laboratory diagnostic techniques is proposed which can assess functional condition of the defense barriers of the upper respiratory tract mucosa in chronic inflammation, estimate impairment of specific and nonspecific resistance. This facilitates choice of optimal scheme of pathogenetic therapy of chronic inflammation in the upper respiratory tracts. PMID- 9752098 TI - [Metabolism and the functional activity of the thyroid in patients with chronic tonsillitis and excess body weight]. AB - Clinical and laboratory tests were conducted in 138 females of reproductive age with chronic decompensated tonsillitis (CDT) with normal and excessive body mass. Improper diet of the obese patients was responsible for specific manifestations of CDT, disturbances of lipid, carbohydrate and mineral metabolism, thyroid function. PMID- 9752100 TI - [The current methodological approaches in assessing nasal breathing function]. AB - The techniques currently used in assessment of nasal respiration with classification and comparative analysis, own experience with maintenance of ATMOS 200 rhinomanometer are outlined. Nasal respiration is best to assess objectively with devices estimating aerodynamic resistance of the nose. The techniques of the anterior and posterior active rhinomanometry (rhinoresistometry) are effective. Acoustic rhinometry is a good adjuvant. PMID- 9752101 TI - [Current methods for assessing mucociliary transport disorders in the diagnosis of chronic rhinosinusitis]. AB - Mucociliary transport of nasal and paranasal sinus mucosa was assessed in 30 healthy subjects and 40 patients with chronic purulent rhinosinusitis. Clinical, biophysical, mathematical, histological and electron microscopic methods were employed. Biophysical investigation comprised life-time TV microscopy of the specimens. Mucociliary transport investigation and sparing sampling of the ciliary epithelium from mucosa surface of the nose and paranasal sinuses were performed by means of a special apparatus. The computer program modelled vibration of the cilia and estimated their movements (total waving frequency, duration of the impact and raising phases, their correlation). The study discovered prominent structural and functional impairment of the mucociliary system manifesting as ciliary dyskinesia and epithelium converting into the stratified squamous one. PMID- 9752102 TI - [Mastering surgical treatment methods for patients with paralytic laryngeal stenosis]. AB - Endoscopic laryngoplasty used in reconstructive surgery of the upper respiratory tracts for paralytic laryngeal stenoses is maximally sparing in relation to laryngeal structures. Mastering of the skill of the endoscopic intervention necessitated application of the device for training surgeons. The device simulates the operative situation of microsurgical manipulations on the larynx. PMID- 9752103 TI - [The surgical rehabilitation of patients with bilateral paralytic laryngeal stenoses]. AB - Effectiveness of surgical rehabilitation was studied in patients with bilateral paralytic laryngostenoses. Reconstructive laryngoplasty with external approach in endoscopic, extralaryngeal and endolaryngeal variants was employed. The examination and treatment of 70 patients who were as a rule of a capable age (85.7%) evidenced for superiority of endoscopic microsurgery. It provides good results and significantly reduces treatment duration. PMID- 9752104 TI - [The acoustic characteristics of the vocalizations of hypoacusic children]. AB - The author analyzes time and frequency characteristics of vocalizations registered in hypoacusis children aged 3-5 months. After the comparison of the acoustic characteristics, utterance duration, formant frequencies of the vocalic utterances, FO frequency changes during vocalization of hypoacusis infants with those of normal infants (literature data), it was determined that infants with impaired and normal hearing do not differ it their vocalizations. PMID- 9752105 TI - The context of condom nonuse among older adolescents and young adults. AB - This exploratory-descriptive study examined the context of condom nonuse among older adolescents and young adults. The convenience sample consisted of 23 freshman university students (18 to 20 years of age) in a rural state. Participants were asked to describe situations when condoms were not used. The responses were content-analyzed using an inductive process. Descriptions (n = 86) were identified and analyzed for common themes. The seven themes were assessment of risks, location and timing of sexual experience, extraneous events, communication about condoms, feelings of the moment, contraceptive responsibility, and relationships between sexual partners. The findings suggest strategies to promote condom use when working with older adolescents and young adults. PMID- 9752106 TI - A comparative study of homeless, previously homeless, and never homeless school aged children's health. AB - The purpose of this cross-sectional study was to compare the mental health, physical health, and healthcare practices of homeless, previously homeless, and never homeless poor school-aged children. The sample was comprised of 134 children who ranged in age from 8 to 12 years. The children participated in health assessments and completed two psychometric tests: the Children's Depression Inventory (CDI) (Kovacs, 1985) and the Revised Children's Manifest Anxiety Scale (RCMAS) (Reynolds & Richmond, 1985). Their mothers completed the Child Behavior Problem Checklist (CBCL) (Achenbach, 1991) and participated in an interview. The homeless (n = 67), previously homeless (n = 30), and never homeless children (n = 37) were similar in regard to their health assessment findings, reported health problems, healthcare practices, and CBCL scores. The proportions of homeless and previously homeless children with CDI scores in the clinical range were significantly greater than the never homeless poor children. The homeless children had significantly higher anxiety scores than the previously homeless and never homeless children. All three groups of children were at risk for physical and mental health problems; however, the findings suggest that school-aged children who experience homelessness may be at greater risk for depression and anxiety than never homeless poor children. PMID- 9752107 TI - Influence of health locus of control and parental health perceptions on follow through with school nurse referral. AB - The health perceptions and health locus of control of 50 parents of students in a school district in New Jersey were investigated in this comparative descriptive study. The study was based on the Health Belief Model and Roger's Science of Unitary Human Beings. Subjects were categorized into two groups, depending on whether or not they followed through with school nurse referrals for their children. Data were collected from each subject to measure levels of health locus of control and health perceptions. The hypothesis stated that parents who do not follow up on health referrals for their children will score lower on health perceptions and health locus of control than parents who schedule referrals for their children. Results revealed no significant differences between compliant and noncompliant parents in health perceptions and health locus of control. Chi square analyses were used to determine the relationships between sample characteristics and participants' responses to the Health Locus of Control scale and the Health Perceptions Questionnaire. PMID- 9752109 TI - PREP and lifelong learning: whose responsibility? PMID- 9752108 TI - Salivary cortisol testing in children. AB - Biological markers can identify links between human biology and human behavior. Cortisol, a marker of hypothalamic-pituitary-adrenocortical axis function, is a useful measure in research. Newer technology involving the measurement of cortisol in saliva is being utilized in research studies. Salivary cortisol measurement is inexpensive and noninvasive and offers many advantages over serum testing. Although there are various methods of saliva collection, it is relatively easy to perform in both infants and children. Salivary cortisol testing may offer a significant measure for pediatric stress, coping, and health research. PMID- 9752110 TI - Project-based learning. AB - Architecture and nursing and midwifery students worked with employers from local trusts on a project exploring design issues for a new maternity unit. Marlene Sinclair and George Brown report on how it was organised, and the benefits for students and employers. PMID- 9752111 TI - The midwifery model of supervision: would it be suitable for nursing? PMID- 9752112 TI - Learning in clinical practice. 3. Facilitating learning. PMID- 9752113 TI - Characteristics of professional relationships. PMID- 9752114 TI - Occupational health nursing. PMID- 9752115 TI - New criteria for the detection, diagnosis, and classification of diabetes mellitus. AB - The diagnosis and classification of diabetes mellitus have been reviewed and revised for the first time since 1979. A precedent-setting recommendation by the American Diabetes Association for routine screening of adults is an effort to identify the estimated 8 million people in the United States with undiagnosed diabetes. Stricter diagnostic criteria will identify more accurately those who are at risk for diabetic complications. Classification and nomenclature have been revised to describe the cause rather than the treatment of diabetes. PMID- 9752116 TI - Pathophysiology of diabetes mellitus. AB - The most common types of diabetes observed in a primary care practice are type 1, type 2, and gestational diabetes. Type 1 diabetes is an autoimmune disease characterized by total destruction of the pancreatic beta cells, whereas insulin resistance, impaired insulin secretion, and inappropriate hepatic glucose secretion characterize type 2 diabetes. Gestational diabetes is similar to type 2 diabetes, but is first diagnosed during pregnancy. PMID- 9752117 TI - Applying nutrition and diabetes recommendations in the nurse practitioner setting. AB - Medical nutrition therapy is essential for successful management in persons with diabetes. This article discusses implementation of the 1994 revised nutrition guidelines for persons with diabetes by the nonregistered dietitian professional, particularly in the nurse practitioner's setting. Major changes in the guidelines are addressed including the liberalization of sucrose, and basic nutrition assessment, referral guidelines, a sample nutrition history, and case studies to assist the nurse practitioner when a registered dietitian is unavailable. PMID- 9752118 TI - The challenge of exercise: practical advice for implementation into clinical practice. AB - Regular physical activity has important physiological and psychological benefits for all people. There are, however, special concerns for people with diabetes that require consideration before recommending an exercise program. Risks associated with exercise include exercise-induced hypoglycemia or hyperglycemia and worsening of long-term complications. Safe participation in all forms of exercise is possible through proper screening and teaching specific diabetes self management skills. PMID- 9752120 TI - Diabetes education and the primary care provider. AB - The problems involved in developing a practical primary care model of individualized diabetes management are discussed. Even though the multidisciplinary team approach is the preferred delivery system for diabetes care, primary care providers who do not have access to a diabetes care team can work with diabetes educators who provide a comprehensive body of knowledge, attitudes, and self-management skills to help people with diabetes make a positive psychosocial adaptation to diabetes. The person with diabetes is the only one in a position to determine what is learned and what is ultimately practiced. PMID- 9752119 TI - Drug therapy in diabetes management. AB - The primary goal in diabetes management is to maintain blood glucose levels as close to normal as possible. Pharmacology is one intervention used to achieve this goal. Medications are used in type 2 diabetes when meal planning and exercise can no longer achieve target glycemic goals. New oral antidiabetic medications and a new insulin analog have increased the number of choices available to practitioners for pharmacological interventions. Understanding the relationship between the pathophysiology of diabetes and the medications' mechanism of action can help practitioners make appropriate therapeutic choices. PMID- 9752121 TI - Acute complications of diabetes mellitus. AB - Blood glucose control is challenged frequently for patients with diabetes mellitus. Illness, stress, and unplanned life events often make glycemic control unstable, which puts the patient at risk for hypoglycemia or hyperglycemia. These acute complications may be mild and managed independently by the patient, or they may be severe, thus threatening life and requiring intensive monitoring. Outcomes are improved with timely recognition of the patient at risk, their symptoms, and appropriate intervention. PMID- 9752122 TI - Chronic complications of diabetes: a creative management approach. AB - The chronic hyperglycemia of diabetes is associated with long-term damage to and failure of various organs, especially the eyes, kidneys, heart, blood vessels, nerves, and, eventually, the individual's sense of well being. With the new diagnostic criteria for diabetes, the incidence of clinical disease is expected to soar. This article will focus on creative management techniques aimed at lessening these complications of diabetes by using the clinical visit efficiently. PMID- 9752123 TI - Caring for feet: patients and nurse practitioners working together. AB - There are many factors to remember when performing a foot assessment on patients with diabetes. To maximize the value of a foot care program, patients need to understand their role in preserving their feet. This article outlines how to perform a logical foot assessment using a mnemonic tool so all parts of the examination will be included. Further actions to consider and educational points to stress are provided for each part of the assessment. PMID- 9752124 TI - Children and adolescents with diabetes mellitus. AB - Nurse practitioners play a vital role in the care of children and adolescents who live with diabetes mellitus. Because of their expertise in combining patient education with patient care, nurse practitioners can provide excellent care both in the primary care setting or as part of a multidisciplinary team. This article is focused on nurse practitioners in the primary care setting and provides up-to date information on the basics of diabetes care. It provides readers with the many aspects of diabetes care including insulin, blood glucose monitoring, and nutrition. PMID- 9752125 TI - The older adult with diabetes. AB - Like their younger counterparts, older adults with diabetes need individualized treatment and educational programs based on personal glucose goals. Although most of the tools and therapies available to younger adults are also appropriate for the elderly, additional considerations and strategies are needed to meet the needs of this population for whom diabetes is a frequent and serious problem. To be effective, the therapeutic approach needs to take into consideration the aging process, other health problems, and the functional, psychosocial, cultural, and educational status of each patient. Along with these considerations, this article provides an overview of the treatment of diabetes for this age group and offers strategies for working with older adults. PMID- 9752127 TI - Waiting list for hospital treatment. PMID- 9752126 TI - Type 2 diabetes in people of color. AB - Type 2 diabetes is a major public health concern for people of color throughout the United States. The prevalence of type 2 diabetes among African-Americans, Hispanics and American Indian/Alaskan Natives is from two to six times greater than that of the US non-Hispanic white population. Rates of end-stage renal disease, amputations, and diabetic retinopathy are also significantly higher. The medical risk factors of familial history, insulin resistance, obesity, history of gestational diabetes, impaired glucose tolerance, and physical inactivity are the same for all populations. The disproportionate impact of diabetes in people of color may be because of an interaction of genetic risk factors and environmental factors. Recognizing the impact of culture in disease management and self-care practices can improve diabetes care. PMID- 9752128 TI - On a short leash? PMID- 9752129 TI - Time to go. PMID- 9752131 TI - Four steps to recruiting nurses. PMID- 9752130 TI - Identity crisis. PMID- 9752132 TI - Access all areas. PMID- 9752133 TI - The Christine Hancock column. PMID- 9752134 TI - Could do better.... PMID- 9752135 TI - Taking care of the carers. PMID- 9752136 TI - Exodus from India. PMID- 9752137 TI - Talking taboos. PMID- 9752138 TI - Life plan. PMID- 9752139 TI - A study of nurse-led shared care for coronary patients. AB - A randomised controlled study of nurse-led shared care of patients awaiting coronary artery bypass surgery led to significant improvements in patients' care management. Here, we summarise the study's findings. PMID- 9752140 TI - Commissioning: the community nurse's role. AB - This article examines the proposal in the White Paper The New NHS (DoH 1997) that community nurses will be involved in commissioning health care and highlights nurses' absence from the commissioning arena in the past. The important contribution nurses can make to commissioning and their interpersonal and communication skills are considered. Factors that may constrain commissioning and strategies to overcome these are also discussed. PMID- 9752141 TI - Therapeutic interaction and mental health nursing. AB - Recent research suggests that interaction between psychiatric nurses and their patients is low. The author of this article argues that this is not a new phenomenon and that psychiatric nursing has failed to grasp its main opportunity for excellence in practice. PMID- 9752142 TI - The challenges of change for learning disabilities nurses. AB - In the past ten years there have been many changes in care provision for people with learning disabilities. This article outlines the changes and discusses the impact they have had on the learning disability nurse's role. PMID- 9752144 TI - Merit awards. PMID- 9752143 TI - Upper gastrointestinal endoscopy: gastroscopy. PMID- 9752145 TI - Dobson's choice. PMID- 9752146 TI - Who's to blame? PMID- 9752147 TI - Peach of a job. Interview by Nick Lipley. PMID- 9752148 TI - Aiming high. PMID- 9752149 TI - In it together. PMID- 9752151 TI - Quality time. PMID- 9752150 TI - Mind control. PMID- 9752152 TI - Will to live. PMID- 9752153 TI - No kidding. PMID- 9752154 TI - A drug-free zone. PMID- 9752155 TI - Kaleidoscope of care. PMID- 9752156 TI - Nutrition for life. PMID- 9752157 TI - Regulating non-nursing healthcare workers. AB - This report looks at a comparative study of the changes in nurse education in the UK and Australia over the past decade. The aim is to examine the impact that a move towards a profession led by degree-qualified nurses might have on the profession's control of the workforce. PMID- 9752158 TI - Causes and treatment of erectile dysfunction. AB - Erectile dysfunction (ED), more commonly known as impotence, is, along with men's health in general, a neglected subject. In this article, the author highlights reasons why ED may occur, the treatments available and why professionals should encourage men to discuss this problem. PMID- 9752159 TI - Primary care: nurse-led telephone triage and advice out-of-hours. AB - In this article the authors report on the evaluation of an out-of-hours telephone triage and advice service in general practice. A computer-based decision support tool was used to guide nurses in the assessment of patients and outcome for calls. Data from the computer system were analysed for all calls over a six-month period. The researchers found that the nurses were able to handle just over half the calls received by giving advice alone. Overall, the service appeared to be remarkably consistent in the decisions taken by nurses, and training and organisational issues to be considered in the future development of the service were identified. PMID- 9752160 TI - Older women's experience of the menopause. AB - Older women are often stereotyped as less productive and less healthy, bemoaning the end of their fertile life and crying out for hormone replacement therapy (HRT). The author of this article challenges these images and suggests that the menopause marks a beginning rather than an end. Nurses should take a more positive approach to women entering the later part of their lives. PMID- 9752161 TI - Anaphylaxis. PMID- 9752162 TI - Male order nursing. PMID- 9752163 TI - Why is success in nursing a boy thing? PMID- 9752164 TI - Primary concern. PMID- 9752165 TI - The RCN's advice. PMID- 9752166 TI - Sex discrimination. PMID- 9752168 TI - Peer pressure. Interview by Adrian O'Dowd. PMID- 9752169 TI - Collaboration between academics and mental health service users can be fraught with difficulties but also deeply rewarding. PMID- 9752167 TI - In your face. PMID- 9752170 TI - Full service. PMID- 9752171 TI - Serving suggestions. The ward sister's view. PMID- 9752172 TI - Serving suggestions. The dietitian's view. PMID- 9752173 TI - Serving suggestions. The doctor's view. PMID- 9752174 TI - Unpalatable options. PMID- 9752175 TI - The rise of Viagra. AB - The old adage 'sex sells' has never seemed so apt. Not since the Pill has a drug made such an impact on the world stage, yet serious concerns surround the introduction and use of Viagra. Jenine Willis investigates. PMID- 9752176 TI - Tackling strokes. AB - The government's green paper sets specific targets for a reduction in the incidence of stroke. Carol Williams and Lynne Kendall look at the nurse's role in prevention and recovery. PMID- 9752177 TI - It's not worth the whistle. PMID- 9752178 TI - Born in the USA. AB - How are newborn babies affected by their mothers' use of drugs nursed in specialist units the USA? A Nursing Times/Birmingham Hospitals Saturday Fund travel award allowed Karena Fellowes to find out. PMID- 9752179 TI - In search of pathways. AB - How can clinical pathways benefit patient care? With her NT/BHSF travel award, Karen Parsley travelled across the world to learn how Australian clinicians implement the system. PMID- 9752180 TI - By giving health care assistants more power and responsibility, nurses are conniving in their own demise. PMID- 9752182 TI - Epidural analgesia: have we got it right? AB - Epidural infusions are being employed more frequently for postoperative and posttraumatic pain. This paper looks at the possible complications involved in the use of epidural analgesia. The evidence concerning possible causes of infection and abscess formation is discussed. The small amount of literature available suggests that the potential for infection is increased when ward-based staff are involved with changing epidural infusions. The potential benefits of ready-prepared solutions for epidural analgesia are considered. PMID- 9752181 TI - Sexual relations in a cold-weather shelter for homeless people. AB - This study was carried out in a cold-weather shelter targeting young people sleeping rough in London. A homeless community mental health team added screening questions to assess sexual and relationship problems. Semistructured interviews were carried out in the shelter with residents not referred to the homeless mental health team. Shelter staff were consulted about their experience of residents' sexual and relationship problems as well as their relationship to substance use. PMID- 9752183 TI - Careers guidance during nurse education: the need for a strategy. AB - This paper reports on an in-depth interview study involving 14 newly qualified nurses and 27 senior health service staff. The findings revealed an unmet demand for career guidance despite the presence of an untapped resource of expertise. PMID- 9752184 TI - The risks of polypharmacy. PMID- 9752185 TI - Prevention of falls in people over 65. PMID- 9752186 TI - Natural solutions. PMID- 9752187 TI - Hands on help. AB - Abdominal massage is a viable alternative to laxatives and enemas in chronic constipation. Ann Richards has developed a massage programme to be taught to informal carers in the community. PMID- 9752188 TI - Beating the burn. PMID- 9752189 TI - With nature's help. PMID- 9752190 TI - A passing phase. PMID- 9752191 TI - Overseas aid: the pitfalls. PMID- 9752192 TI - Family fortunes. PMID- 9752193 TI - New perspective for an old joke. PMID- 9752194 TI - Green cycle starts here. PMID- 9752195 TI - Cruel to be kind. PMID- 9752196 TI - Appreciating the benefits that eugenics could bring to the human race. PMID- 9752197 TI - Nancy the Named Net Nurse, professional autonomy, a documented knowledge base. PMID- 9752198 TI - The government's payout to the NHS. PMID- 9752199 TI - What's your poison? PMID- 9752200 TI - User-friendly. PMID- 9752201 TI - Drink drive. PMID- 9752202 TI - Beyond the fringe. PMID- 9752203 TI - The beast inside. PMID- 9752204 TI - Down with demarcation. PMID- 9752205 TI - Coffee break. Interview by Pat Anderson. PMID- 9752206 TI - Face to face. Interview by Eileen Fursland. PMID- 9752207 TI - Postherpetic neuralgia: a care study. PMID- 9752208 TI - Who becomes a mental health nurse? AB - Mental health nurses come to the profession with a diverse range of life experiences. Gary Hickey and Cheryl Kipping describe a recent survey and its implications for recruitment and retention policies. PMID- 9752209 TI - Managing deep vein thrombosis at home. PMID- 9752211 TI - Flying start. AB - The transition from a secure college environment to the unfamiliarity of an operating theatre can be bewildering. But with proper preparation and a good mentor, says Fiona Martin, it can also be inspiring. PMID- 9752210 TI - A world of difference. AB - After years of striving, theatre nurses' perioperative role has finally been recognised. The changes this brings, predicts Paul Wicker, will be wide-ranging. PMID- 9752214 TI - [Elections 1998. How do the parties want to shape the nursing professions?] [In Process Citation] PMID- 9752213 TI - [Whom should you trust?] PMID- 9752212 TI - Operating profits. AB - Training student nurses for the rigours of the theatre is no easy option. So why do mentors bother? Dina Plowes may have found the answer. PMID- 9752215 TI - [Nursing care needs specialists--specific tasks for professionals as compared to aides] [In Process Citation] PMID- 9752216 TI - [Decubitus ulcers and drugs. No matter what their composition, drugs are not suitable for the prevention of decubitus ulcers. Modern therapeutic methods should be used instead] [In Process Citation] PMID- 9752217 TI - [A nocturnal explantation--feelings of an operating room nurse] [In Process Citation] PMID- 9752218 TI - [Accidents with oxygen armatures] [In Process Citation] PMID- 9752219 TI - [Interhospital and interfab 1998] [In Process Citation] PMID- 9752220 TI - [The market and the inferno] [In Process Citation] PMID- 9752221 TI - [Nursing school before the closing: we cannot do without education]. PMID- 9752222 TI - [Reduced numbers of visitors and exhibitors at the Interhospital/Interfab 1998: the organizer sees the course of the fair in a positive light]. PMID- 9752223 TI - [The federal minister for health, Seehofer, states: "The savings lemon in German delivery of health care has been squeezed dry"]. PMID- 9752224 TI - [Necessity to deal with the consequences of longevity: geriatric patients often suffer from malnutrition]. PMID- 9752225 TI - [Anesthesia and intensive care: maintaining the activities and existential experiences of life]. PMID- 9752226 TI - [Relaxed sleep--how to achieve it? There are various possibilities to avoid tablets]. PMID- 9752227 TI - [Observing the patient with open senses. Five senses--or are there more?]. PMID- 9752228 TI - [Foot reflex massage in nursing: it can be of help in including family]. PMID- 9752229 TI - [With propolis to new health: natural healing with propolis]. PMID- 9752230 TI - [Treatment of decubitus ulcers/wound management: a dry wound is a dead wound]. PMID- 9752231 TI - [Public health as a preventive effort: the aim is prevention of health risks]. PMID- 9752232 TI - [Quality assurance in times of cut-backs 2. Trust among the co-workers creates trust among the patients]. PMID- 9752234 TI - [What remains from the legislative marathon for hospital financing? It does not pay to exceed the agreed-upon services]. PMID- 9752233 TI - [Guide for efficiency analysis of a patient documentation system: the first step is the formulation of the targets]. PMID- 9752235 TI - [The caregiver makes the decisions--or does he? 1. Nurses are between 2 fronts]. PMID- 9752236 TI - [50 years of Pflegezeitschrift: "how the floor nurse gets cracks"]. PMID- 9752237 TI - [Primary care at the protestant Amalie Sieveking Hospital. Report of experiences after 2 years of work on the project]. PMID- 9752238 TI - [Nursing care syndrome]. AB - This paper has as a principal goal to describe the 'nursing care syndrome' that appears while nursing care is provided. From the conceptualization of 'nursing care syndrome' besides other aspects, it points out the difference between the latter and the iatrogenicities as well as between the clinical syndromes and the nursing care ones. PMID- 9752239 TI - [Expression of sexuality of the hospitalized patient and the nurses' strategy in care]. AB - Nursing is the science and the art of caring of the human being and it has to consider that this human being, in an integrated way, has psychological, social, biological and spiritual needs. We understand sexuality as an important dimension of the human being which requires attention considering a holistic assistance. From these principles, we have attempted to research next to a Rio de Janeiro city school hospital nurses, how these professionals deal with the sexual body and the hospitalized client sexuality expression in the context of the daily care provided for him. We have performed 10 non-structured interviews and proceeded to the analysis of their speeches contents. The analysis showed that the nurses when providing direct care to clients, regarding his sexuality, they experience difficulties that must be overdrawn so they can perform care. During the results follow-up, the nurses refer to make use of 'strategies' that hide the client's sexuality expression and demonstrate to be fully unprepared to deal with such situations. Such 'strategies' show that in the disciplined hospital space, there is no overture for the understanding of a body with historical, social and cultural scars; there is no room to deal with questions which take to emotion and human subjectiveness realms. We do believe that there is a need of treating the hospitalized client sexuality in critical and contextualized discussions linked to social and cultural fields. These discussions must be done both in care praxis to client and professional degree. PMID- 9752240 TI - [Preoperative orientation: comprehensive analysis of the patient's point of view]. AB - In this study we approach the issue preoperatory orientation which we understand to be a vital practice in the field of Nursing also allowing the identification of same features at clients perception regarding the information achieved before operation. It is also suitable to recognize the failure of this procedure in the daily work at surgical units which upsets us with the absence of a guideline for preparing patients to surgical acts. Facing this need, we have decided to deeply investigate this issue to achieve the dimension, the description and analyzing the trajectory follow-up started in the first contact with the phenomenological approach. Thus, we make use of our patient operatory phase through his talk according to MARTINS & BICUDO's reference. So, this research aims at presenting the knowledge feature achieved through patients talk, its meaning and repercussion onto the individual plane as well as identifying what should alone improve it. That is the way phenomenological method questions the need of a preoperatory orientation which links us to theme dimension. In our experience of guiding patients to cope with their surgical intervention, we open a space that allows us to intervene under the perspective of conducting patients to recognize their existential dimension of experiencing an operation. PMID- 9752241 TI - [Group process in nursing: possibilities and limits]. AB - This paper describes our study trajectory and reflection on the use of groups in Care. Through a qualitative perspective we aim at identifying, under the interviewees perception, the aspects which motivate them for this activity, their learning sources and their group concrete experiences relevant topics. We have verified that nurses recognize the therapeutic value of this work which promotes them adequate structural and physical conditions. However, it has been observed that one of the limiting factor for this activity is the difficulty of dealing with group situations which reveal the nuances of human feeling indicating that the group coordinator nurse besides needing theoretical resources, he also needs to exercise his self-knowledge in order to provide a group relationship and environment able to optimize this activity therapeutic value. PMID- 9752242 TI - [Alteration of oxygen saturation during endotracheal aspiration]. AB - This study verified the occurrence of oxygen saturation alteration during endotracheal aspiration. Patients were adults aged from 17 to 70, admitted due to either neurologic diseases, neuro or abdominal surgical affections. All were devoid of pulmonary pathology. Suction endotracheal catheters numbers 12, 14, and 16 were used. Variation in oxygen saturation was discrete and similar in the use of the three experimental catheters but, for practical purposes, numbers 12 and 14 proved to be easier to operate and more comfortable for the patients. PMID- 9752243 TI - [Are the roles of the nurse specialist and the master of health services the same?]. AB - This study aimed at identifying the role of the nurse with a graduate degree at the service level based on nurses and service directors expectations in health care institutions. A total of 278 nurses and 25 service directors responded to a questionnaire and an instrument was designed to measure their expectations towards the nurse with a postgraduation degree. The results indicate that nurses and service directors hope nurses with a postgraduation degree to be highly qualified to provide care. While service directors express a slight difference in favor of both the master degree and the deep knowledge of the specialty, nurses expectations in relation to it, are a little greater in all aspects. Service directors priorize refresh courses for nurses while nurses priorize specialization courses. The study has implications for a deeper discussion of these professionals functions in service and for the objective of postgraduate education programs in the region. PMID- 9752244 TI - [Pleasure and pain at work: contribution to the organization of nursing activities]. AB - This paper is the synthesis of the author master program in administration thesis. It studied the pleasure, suffering and work relationship and searched to establish the implications of work itself, work organization and offered work conditions in the feelings of pleasure and suffering of nursing workers. PMID- 9752245 TI - [Representation of disease among postgraduates]. AB - This study analyses the concepts of disease in a different historical moment and taking LAPLANTINE's anthropological background, analyses, in a postgraduate team, their disease representation as own experience and as professional experience, in four ethnological pairs proposed by the author. PMID- 9752246 TI - [Reasons for personnel turnover in a private health facility]. AB - This study was done in a Sao Paulo city hospital with 54 nurses who required for dismissal from January, 1995 to January, 1996. All of them at the dismissal interview provided information on how long they had worked for the hospital, reasons for leaving and their frank opinions about the hospital itself, nurse department directors and personal suggestions. The study goal was to evaluate the reasons for high turnover of institutions nurses in order to draw adequate administrative intervention strategies. An exploratory study of retrospective character with data quantitative qualitative analysis has been used. Data analysis considered that 74% of the nurses had left the institution in 12 months but 35.3% until the third month. The nurses emphasized work conditions specially the staff, benefits and interpersonal communication. The results came towards our perception on the institution nurses turnover causes. PMID- 9752247 TI - [Characterizing research activities from the time of their introduction: nurses' reports]. AB - My interest in developing this study came from a previous work (Ricci, 1994) about scientific research meaning for Pharmacy and Nursing students. After its conclusion, it was necessary to know scientific research historical evolution pointing out the epoch and factors which contributed for its development. Aiming at reconstituting Nursing research historical evolution, trying to find the moment of its development in the profession, identify the reactions research introduction in the course and the assistance in opinions of docent nurses. For this, a semi-structured interview has been used. The population for this study was of nurses who lived the 60's and 70's. At first I obtained a list of nine names of nurses, but according to the theoretical reference, data became saturated along the interview, therefore the final sample was of only four nurses. The definition of generated themes from nurses speeches analysis was presented. The analysis allowed the discovery of two themes: STARTING ON RESEARCH and NEEDING REFRESH. The data revealed the difficulties that the docent nurses experienced by the time of research and postgraduation introductions. It has been observed that the arguments used by those nurses are the same used today by assistance nurses who want to investigate (Cassiani, 1994). However, the main difference lays on the fact that for the docent nurses there was a demand and for the assistance ones there is an internal need that not always is appraised and stimulated in service. PMID- 9752248 TI - [Classification of nursing practices in public health in Brazil]. PMID- 9752249 TI - [Reading nursing journals]. PMID- 9752250 TI - [International health: a new challenge to nursing education]. AB - A survey was conducted among 110 Schools of Nursing in USA and 5 Schools in Latin America and the Caribbean (LAC) to identify the international health (IH) component in nursing education, practice and research. A significant part of U.S. schools and all 5 LAC schools have international activities, and this interest has started basically in the last 5 to 10 years. There was difference in the structure of IH activities among U.S. and LAC nursing schools, but they were similar in the type of support offered to IH initiatives. IH content in nursing education among U.S. schools was related to culture, health systems and community health; in LAC schools, IH content was related to health promotion, health policy and strategies and nursing perspectives. U.S. and LAC schools with international activities have only 10% of their faculty and students involved with IH initiatives. The nursing schools still lack courses and activities that the Schools of Public Health have implemented to deal with IH. The article observes areas that need to be strengthened so that nursing professionals can expand their leadership roles in research and practice in international health. PMID- 9752251 TI - [The teaching of physical examination in undergraduate nursing schools in the city of Sao Paulo]. AB - The present study is an exploratory survey attempting to get the point of view of the faculty of subjects considered responsible by teaching physical examination in the nursing undergraduate schools of Sao Paulo city that were selected for this study. The goals were: to detect the situation of teaching of propaedeutics basis; to identify factors that make them difficult; and to identify the recommendations of the faculty for the improvement of teaching and its implementation. The population studied was formed by 39 faculty of the subjects considered responsible by teaching physical examination. Results show us there is not a specific subject to teach physical examination, although it is taught mainly in Fundamental Nursing; faculty are not entirely prepared to teach physical examination. To improve teaching of physical examination, the faculty suggest enhancing their knowledge and skills as well as to create a specific subject to teach physical examination with sufficient hours according to the content. PMID- 9752252 TI - [Follow-up of nurses who have finished the specialization courses in intensive care nursing]. AB - The purpose of this study was to analyse the contribution of the Extension Courses in the Intensive Care held at the Nursing School from USP, with the aim to: 1) Follow up nurses in their work at the ICU. 2) Evaluate the contribution offered by the course. The population was formed by 38 nurses, considering that 60.5% didn't work in the ICU anymore mainly because they assumed another function in the institution and had several private problems. Regarding the contribution offered by the course, the main evaluation was the acquirement of knowledge (91.7%). However, 74.2% of the nurses declared the course didn't contribute to the increase of their wage, what do not impair them from investing in their professional improvement. PMID- 9752253 TI - [Continuing education of the intensive care units nursing staff in the city of Sao Paulo]. AB - This study is part of a project about ICU's characteristics in Sao Paulo city. This article describes the continuing education programs for nursing staff. 43 ICUs were analysed and the questionnaire answered by the ICU nurse coordinator was used to collect data. Results showed that 34 (79.1%) of the ICUs have initial program for training each nursing staff category and 18 (41.9%) had regular continuing education program focusing primarily on nursing procedures and routines and the update in pathologies. Continuing education programs are developed primarily by ICU's nurses. 50.2% of the nurses answered that they attended specialisation/extension courses too, in Medical Surgical Nursing or in other areas or in both areas. Due to this result and the development of the ICU's nurses as specialists, some suggestions are presented to improve the continuing education program in these Units. PMID- 9752254 TI - [Special education in stoma therapy in Brazil 1990-1995]. AB - The authors show the trajectory of the Brazilian Stomal Therapy Courses since their beginning in 1990 until 1995. Firstly they point out the changes of their contents initially based on medical and biological fields. Nowadays they also involve issues in the other Enterostomal Therapy fields-wounds and incontinence besides discussions and reflections on the principles of organization and creation of care programmes, services and protocols as well as on the Stomal Therapy as a specialty in Brazilian nursing, society and politics. The 75 students' expectations, perspectives and evaluation related to the courses are presented, showing positive tendencies. The author finishes the study considering the importance of the evaluation process towards the improvement of the course according to the new challenges and discussions about nursing specialties in our country. PMID- 9752255 TI - [Public health literature: teaching of a new look in the health-illness process]. AB - This paper presents the use of Brazilian literature in public health nursing courses at the University of Sao Paulo at Ribeirao Preto College of Nursing. The goal is a preliminary report about experiences that intend to keep the students in touch in order to wide their view of reality, in which relationships between the men and health-illness process are established in the complexity and singularity of human being (ill and life) in each place and time. In the first stage, this experience is based on New History conceptual framework (Ecolle des Annale), in the complexity's paradigm (MORIN) and circularity concept and sign's paradigm (GINZBURG). PMID- 9752256 TI - [Breast feeding: importance of supportive counseling to the working mother]. AB - This descriptive, exploratory, retrospective and transversal investigation tries to answer the following questionnaire: What is the impact that the Support Consultation to the working mother--Diagnosis Center of the Pontifical Catholic University of Chile--has on breast-feeding prolongation? The population that has been studied is formed by 82 mothers attended in the Support Consultation during March '95/September '95 period and from which a number of 30 people were taken as a sample. The impact of this consultation was evaluated with an instrument employed during an interview to each mother. Data were statistically analyzed with EPIINFO, the Kaplan-Meier survival method and the Mantel-Haenszel test to compare curves of survival. During data analysis authors found that mothers are mainly young adults, stable couples, first-time mothers, with technical and/or professional educational level, chiefly working as office clerks with full-time jobs and having a significant difference between existing minimum salary and the maximum one they earn. The results of this investigation lead us to the conclusion that mothers obtained an exclusive breast-feeding and ideal weaning age. The power of resolution of the Consultation--according to mothers--was satisfactory. The support given to the mother after her reincorporation to work is the most significant intensifier factor in relation with the increase in the probability of keeping on breast-feeding. In conclusion, the Consultation has good impact. PMID- 9752257 TI - [Breast feeding, a nature-culture hybrid]. AB - This article consists in an essay that aims, through a review of bibliography, to discuss the relation between a proposal of Bruno Latour about nature-culture and breast feeding. According to his proposal, having as reference an epistemological view, there is an attempt to establish dialogue between different scientific achievements with a view to contribute to the improvement of this subject in the field of public health. Firstly, the paradox of weaning is found in the scenery of knowledge and practices of breast feeding. The analysis is based on the relationship nature-science and culture-society, which are present in the process of breast feeding. Finally, the discussion has the aim of dealing with breast feeding as long as a hybrid between nature and culture, starting a new issue to the continuity in the debate about the thematic in question. PMID- 9752259 TI - [Fluid volume deficit: profile of defining characteristics in patients with burns]. AB - The present study aimed at describing the profile of defining characteristics in patients with the nursing diagnosis "Fluid volume deficit" related to active loss of fluid secondary to burns. Data were collected by means of a tool, containing 29 possible defining characteristics of this diagnosis. Seven nurses, that worked at the Burnt Unit for at least five years ago, provided opinions about the degree to which each defining characteristic is indicative of this diagnosis. Nurses rated each defining characteristic of diagnosis being tested on a scale of zero to one. The results confirmed all, except one (increased body temperature) defining characteristics presented by NANDA for this diagnosis and indicated 10 new defining characteristics. PMID- 9752258 TI - [Measurement and assessment of postoperative pain: a short review]. AB - How to measure pain is a great challenge to those who desire to control adequately such a complex experience. Standardized instruments that take into consideration the patient's own account, have been developed in order to make such a task easier. In this article we carry out a revision of the instruments used mostly for measuring postoperative pain, and we point out some of the advantages and disadvantages. We emphasize the need for specific research focusing on the measurement of surgical pain, taking into consideration the multiple dimensions of a painful experience. PMID- 9752260 TI - [Disinfection of medical and surgical equipment: efficacy of chemical disinfectants and water and soap]. AB - In this experimental study we compared the previous descontamination efficacy of the medical surgical materials by the use of chemical disinfectants and the mechanical cleaning with water and soap, as well as verified the organic material interference in these procedures. To carry out this study, we used surgical pincers under contamination with: Staphylococcus aureus ATCC-6538, Salmonella cholerae suis ATCC-10708, and Pseudomonas aeruginosa ATCC-15442 in presence and absence of organic matter (fetal bovine serum). The following treatments: glutaraldehyde 2%, sodium hypochlorite 1%, hydrogen peroxide 6%, alcohol 70% and the mechanical cleaning with water and soap were compared with eight repetitions in a total of 480 observations. In the described conditions, the disinfectants had a good efficacy in the previous descontamination of the medical surgical materials and a less inactivation by the organic material. The mechanical cleaning with water and soap showed a reduction of the microrganism to safe levels, considered adequate for previous descontamination. PMID- 9752262 TI - [The Ribeirao Nursing School and its contribution to the development of nursing]. PMID- 9752261 TI - [Declarations by registered nurses from the 1980s: an aid to the understanding of nursing today]. AB - This study is part of a more extensive project that aims to rescue significant aspects related to the evolution of nursing care from 1950s until 1990s. This study was developed using the technique of oral declaration by active and retired registered nurses, in the context of a school-hospital from the interior of the Sao Paulo State. The present study particularizes the outcomes regarding to 1980s. As result becomes evident the undertaken effort by nurses in the struggle for profession's recognition and prestige; intense and deep transformations related to nurse's new roles as leadership of the nursing staff and members of the medical team. PMID- 9752263 TI - [Center for post-anesthetic recovery: observation, analysis and comparison]. PMID- 9752264 TI - [Have they discovered the origins of aneuploidy in tumors?]. PMID- 9752265 TI - [APHS and celecoxib, the new aspirins have arrived]. PMID- 9752266 TI - [Histone deacetylase and retinoblastoma protein]. AB - The balance between cellular proliferation and differentiation is strictly controlled in the cell and the deregulation of this balance can lead to tumour formation. The tumour suppressor protein Rb plays a key role in this balance essentially by repressing progression through the cell cycle and thereby it blocks the cell in G1 phase. Rb represses S phase genes through the recruitment of an enzyme which modifies DNA structure, the histone deacetylase HDAC1. The Rb/HDAC1 complex is a key element in the control of cell proliferation and differentiation. Moreover, this complex is likely to be a target for transforming viral proteins. PMID- 9752267 TI - [Metastatic carcinoma of unknown origin]. AB - Carcinomas of unknown primary site are microscopically confirmed metastatic epithelial malignancies with no identified primary site at the onset of treatment. Their incidence is about 5% among all cancer patients. They represent a group of heterogeneous tumors with low chemosensitivity and poor outcome: the overall median survival is about 6 months. The search for primary tumor should be limited to the identification of subgroups of patients requiring specific therapies: 1) patients with cervical lymph nodes containing squamous carcinoma; 2) women with axillary lymph nodes containing adenocarcinoma; 3) women with peritoneal adenocarcinomatosis; 4) young men with poorly differentiated carcinoma of the midline; 5) patients with neuroendocrine metastasis. Other patients are to be managed with symptomatic procedures and possibly chemotherapy. Simple prognostic factors such as performance status, histology and serum levels of alkaline phosphatase may help the physician to select treatment strategies with the aim of preserving an optimal quality of life. PMID- 9752268 TI - [Do the prevention and treatment of deep venous thrombosis have specificity in the cancer patient?]. AB - An increased incidence of deep vein thrombosis is reported in cancer patients as compared with general population. Several risk factors for deep vein thrombosis have been identified: venous stasis, direct invasion of venous wall by tumor, and hypercoagulability state by inadequate secretion of procoagulant activities. Reports have suggested that chemotherapeutic agents and hormonal treatment may contribute to this risk. Few papers are available concerning the prophylaxis and curative treatment of deep vein thrombosis in cancer patients and no consensus has been reached yet. This predisposition for deep vein thrombosis should be taken into consideration for perioperative prophylaxis. Efficacy and safety of heparin and antivitamin K in the curative treatment of deep vein thrombosis are discussed but can not be accurately evaluated because of diversity of clinical presentations and mechanism of activation of coagulation. Prospective studies are necessary. PMID- 9752269 TI - [Targeting the gene of glucose metabolism for the treatment of advanced gliomas]. AB - Loss of chromosomes is a recurrent event in cancer. Chromosome-10 losses occur with tumor progression and characterize advanced gliomas. This chromosome carries many genes involved in glucose metabolism. Hexokinase (HK) gene is located on chromosome-10. Hexokinase enzymatic activity is decreased in glioblastomas. Hexokinase enables glucose entry into glycolysis and is critical for these highly glycolytic tumors. These enzyme is either free in the cytosol or bound to the mitochondrial outer membrane. Disturbance of HK binding to mitochondria by lonidamine led to inhibition of cells and xenografted-glioma growth. Hexokinase bind to a mitochondrial porin which involved peripheral benzodiazepine receptors. Inhibition of HK and peripheral benzodiazepine receptors by lonidamine and diazepam led to synergistic antitumoral activity in xenografted gliomas. Co inhibition of these two receptors will lead to a decrease in glycolysis, often elevated in these tumors, without modifying energetic metabolism of normal cells. PMID- 9752270 TI - Genetics of familial atypical multiple mole-melanoma (FAMMM) syndrome in The Netherlands: how far have we come? AB - By the genetic localization of the first melanoma susceptibility gene on chromosome 1p we thought that the puzzle on familial melanoma families would soon be solved. Now, almost fifteen years later we have learned that inherited melanoma is not a simple genetic disorder and that multiple genes, modifying genes and environmental factors might be involved. This paper outlines the current understanding of the genetics of melanoma and the relationship to atypical nevi based on more than ten years of genetic analysis in the Dutch familial atypical multiple mole-melanoma (FAMMM) syndrome families. PMID- 9752271 TI - Improvement of the Cockcroft-Gault equation for predicting glomerular filtration in cancer patients. AB - In order to determine the morphological and biological covariables which better predict the glomerular filtration rate in cancer patients, we performed the present study in a population of 123 patients (78 men, 45 women) with various tumor types; 55 of these patients had previously received cisplatin, and 12 had undergone unilateral nephrectomy. The 51Cr-EDTA plasma concentration versus time data of 80 patients were analysed according to a population pharmacokinetic approach by using the Nonlinear Mixed Effects Model (Nonmem) program. The best fit for 51Cr-EDTA clearance estimation was given by the following formula: [formula: see text] (with ABW for actual body weight in kg, age in years, sex = 0 if male and sex = 1 if female, and Scr for serum creatinine in mumol/l). Actual body weight was the most predictive morphologic parameter, and the adjustment was not improved by taking into account the ideal body weight. The GFR of patients previously treated with cisplatin was 18% lower than that of untreated patients age for age. However, this covariable was not present in the final model because it was redundant with other covariables, likely Scr. The formula was then prospectively evaluated with the data of 43 other patients. The mean (+/- SD) ratio between GFR predicted according the Nonmem formula and the observed GFR was 0.95 +/- 0.23 which did not differ significantly from unity. Conversely, the mean ratio between creatinine clearance calculated according to the Cockcroft-Gault equation and the observed GFR (0.86 +/- 0.21) differed significantly from unity. This study shows that in cancer patients the formula to calculate GFR drawn from Nonmem analysis is more accurate than the Cockcroft-Gault equation. However, an accurate determination of GFR requires specific techniques as 51Cr-EDTA clearance investigation. PMID- 9752272 TI - [Psychosocial impact of the first oncogenetic consultation in a familial context of cancers of the breast and the ovary]. AB - The purpose of the oncogenetic consultation, is to respond to persons who wonder about their risk of developing a tumour and wish to learn about ways of prevention and detection. The object of this investigation is (1) to study the impact of a first consultation in genetics on the quality of life and the psychology of women with a family history of cancer, and women with or without family antecedents but who suffered themselves from breast cancer at an early age (< or = 35 years); (2) to evaluate their risk perception and their comprehension of the information after the consultation. The study was performed on 200 women attending a first consultation at Institut Curie. Fifty-nine of them had no cancer. Among the 141 consultants with cancer, 54 had developed breast cancer at an early age (< or = 35 years). Their quality of life, their psychological state and their knowledge of the risk of breast tumour were evaluated before consulting and 6-8 months afterwards. Before consulting, their quality of life was altered in some (non-relational) fields but their psychic condition was relatively maintained. Six-nine months after consulting, their quality of life had not deteriorated, but tests results on their psychological state were not as good. The initial psychic condition, itself related to the medical status of the consulting women, was the most significant predictor of their quality of life a few months after their first genetics consultation. After consulting, the women expressed satisfaction with the information delivered by the genetician and their appreciation of tumour risks was improved. PMID- 9752273 TI - [Assessment of the quality of life in the surgery of cancer of the esophagus: critical review of the literature]. AB - The surgery of the oesophageal carcinoma is subjected today to a renewed interest. In the objective to draw recommendations for the clinicians wishing to include quality of life measures in clinical trials, it seemed pertinent to us to critically review existing studies. The review of the literature, presented here, is based on a bibliographic research using Medline database (January 1986 December 1996). A precise methodology was adopted to read the eighteen articles retained and the psychometric instruments used in each study were listed and classified. Nine studies considered the issue of quality of life but failed to measure the outcome or used not validated psychometric instruments and 4 others used only indicators of physical performance. Only five studies assessed quality of life using instruments with known psychometric properties (Quality of Life Index of Spitzer, Rotterdam Symptom Checklist and EORTC QLQ-C30 questionnaire) more often associated to ad hoc dysphagia scales. This review of literature shows that the inclusion of validated quality of life measures in the surgery of oesophageal carcinoma is very recent and that it presents some difficulties inherent to the lack of specific tools. The EORTC QLQ-OES24, specific of oesophageal cancer, being not yet validated in french, the authors recommend the use of the EORTC QLQ-C30 jointly with ad hoc dysphagia scales. PMID- 9752274 TI - [Double-hybrid system in yeasts]. PMID- 9752275 TI - [Postoperative radiotherapy improves the carcinologic results of tumor resection for galactophorous carcinoma in situ of the breast]. PMID- 9752276 TI - [Upon survivin, the neuroblastoma does not longer live...]. PMID- 9752277 TI - [At the front line of malignant melanoma, great maneuvers: to cut the road to Bcl2 and "attack with chemotherapy"]. PMID- 9752278 TI - [From dendritic cells to the exosomes that they secrete: a decisive advance for anticancer vaccine therapy?]. PMID- 9752279 TI - [When p19ARF finds a partner or new "dangerous liaisons"]. AB - In man, the MTS1 (multiple tumor suppressor 1) locus has been located on chromosome band 9p21 and encodes two unrelated genes, p16INK4a and p19ARF that both act, through different mechanisms, as cell proliferation inhibitors. The inhibitory mechanism of p19ARF has begun to be elucidated by the finding that the protein has the ability to bind the mdm2 oncoprotein. mdm2 is implicated in the degradation of p53 and its functional inactivation. While this result provides an opportunity for understanding the tumor suppression role of p19ARF, it allows to define a new cell cycle regulatory pathway, the p19ARF-p53 pathway. In this respect, MTS1 is unique in its ability to control two crucial pathways that regulate the cell cycle. For this reason, it will be crucial to investigate the status of p19ARF in a variety human tumors. PMID- 9752280 TI - [Signal transduction from the membrane to the nucleus: variations on common themes]. AB - During their life, cells are exposed to a wide variety of extracellular stimuli and have to develop appropriate biological responses. Signal transduction from the plasma membrane, which is in contact with the extracellular environment, to the nucleus, where gene expression is achieved, thus represents a fundamental process for the development and maintenance of life in organisms. Signalling pathways are extremely diverse and range from direct strategies, such as the steroid hormone receptor and JAK/STAT (signal transducers and activators of transcription) pathways, to multi-step strategies, such as the NF-kappa B (nuclear factor kappa B), PKA (protein kinase A) and Ras/MAPK (mitogen-activated protein kinase) pathways. In order to modulate gene expression, all these pathways must ultimately achieve nuclear localization. The mechanisms by which these varied signalling components cross the nuclear envelope are equally as diverse. However, despite the variety of the means used, cells have adopted several common themes for signal transduction, particularly interaction between proteins as a mean to transport the signal and phosphorylation as a post translational modification carrying information. Finally, all signalling pathways have been conserved throughout evolution, inghlighting their advantage for cells. In mammals, proteins that participate in signal transmission represent a frequent target for mutations leading to tumor development. Unraveling signalling pathways thus represents an important step in the fight against cancer. PMID- 9752281 TI - [Evolution of the surgery of cancer of the breast]. AB - Multiple technical and strategical improvements have modified surgical management of breast cancer. Screening mamographies are able to focus small tumors in which excision eventually after percutaneous biopsy can necessitate two surgical procedures. Reduction of axillary dissection in these small lesions can benefit from sentinel node biopsy technique. Immediate or delayed breast reconstruction reduces the mutilation of the mastectomies with their psycho-social disaster. Technically these reconstruction requires a multidisciplinary approach and psychological support. These surgical evolutions in parallel with other therapies for breast cancer developed with two major points: necessity of a good local control for cure, multidisciplinary approach which makes surgeons reals oncologists. PMID- 9752282 TI - [Standards, Options, and Recommendations for the management of patients with malignant mesothelioma of the pleura. Federation Nationale des Centres de Lutte Contre le Cancer]. AB - The "Standards, Options and Recommandations" (SOR), started in 1993, are a collaborative project between the Federation of the French Cancer Centres (FNCLCC), the 20 French Cancer Centres and specialists from French Public Universities, General Hospitals and Private Clinics. The main objective is the development of clinical practice guidelines to improve the quality of health care and outcomes for cancer patients. The methodology is based on literature review and critical appraisal by a multidisciplinary experts group, with feedback from specialists in cancer care delivery. The objectives are to develop a clinical practice guideline with definitions of Standards. Options and Recommendations for the clinical care of malignant pleural mesothelioma. Data have been identified by literature search using Medline (1966-June 1997) and personal references lists. The main criteria considered were incidence, risk factors, pronostic factors and efficacy of cancer treatment. Once the guideline was defined, the document was submitted to 40 independent reviewers for peer review, and to the medical committees of the 20 French Cancer Centres for review and agreement. The results are: 1) systematic assessment of (professional) exposure to asbestos is based on a standardized interrogatory, completed by specific consultation for professional disease; 2) diagnostic and clinical staging is based on multiple biopsies under thoracoscopy and thoracic scanner; 3) there is no indication for extemporaneous examination, immunocytochemistry should use cytokeratine, EMA, vimentine, ACE, Leu-M1; 4) clinical care: the recommended staging classification is the IMIG (International Mesothelioma Interest Group) classification; 5) validated, independent pronostic factors are stage of disease patient's functional status and histologic type (i.e. epithelial lesions are of better prognosis); 6) treatment is based on symptomatic and palliative treatment options. Anticancer treatments (surgery, chemotherapy, immunotherapy, radiotherapy) did not show significant improvement of survival. The inclusion of patients in clinical trials is recommended. PMID- 9752283 TI - [Granisetron (per os) compared with ondansetron (per os) in the prevention of nausea and vomiting induced by mildly emetogenic chemotherapies. Groupe de Recherches en Cancerologie du Nord]. AB - It is a randomised cross-over multicenter study comparing the efficacy and the tolerance of granisetron (Gra) 1 mg and ondansetron (Ond) 8 mg, oral, given during two consecutive cycles to 188 naive patients scheduled to receive a moderately emetogenic chemotherapy. The antiemetic treatment is given one day per course, 1 hour before chemotherapy and the second administration from 8 to 12 hours after the beginning, during each of the two cycles; alternatively according to the randomisation. Five criteria are assessed; nausea (ordinal and visual analogic scales), emeric episodes (vomiting orland retching), complete response (minor or no nausea, no emetic episode and no rescue treatment), patient preference and tolerance. The intent to treat analysis showed no significant difference at cycle 1 between Gra and Ond; at cycle 2, there is no significant difference in the number of emetic episodes; for the prevention of nausea, the ordinal scale shows a significant difference (p = 0.028 in favour of Gra at day 1 (D1) but not from D2 to D5. Gra induced more complete response than Ond at D1 (p = 0.028), but not from D2 to D5. The cross-over study did not show any period or order effect, whereas a treatment effect on Ond was significant in favour of Gra (p = 0.01). There is no significant patients preference in favour of Gra or Ond. In conclusion, Gra was more efficient in preventing nausea and obtaining complete response on the first day of treatment, significantly at the second cycle. Both Gra and Ond had a good antiemetic activity for moderately emetogenic chemotherapy with complete response rates always over 50% on day 1; delayed emesis remain less weli controlled. PMID- 9752284 TI - [Influence of knowledge relative to cancer on prevention behavior in the population of Bas-Rhin]. AB - The tendency today is to make the population more aware of health in order to encourage their use of prevention activities. The aim of our telephone survey which was performed on a representative sample of the population (1,010) in the Bas-Rhin department in France, was to study the relationship between the level of knowledge of cancer and the use of screening techniques to prevent it. Eighty-two percent of the males and 78% of the females surveyed, show a profound understanding of cancer in general, the associated risk factors and its prevention. The level of awareness of males did not influence a specific behaviour (p = 0.68) towards cancer prevention. It was shown that in smokers, the more they smoke, the more they underestimate the chances of cure from all categories of cancer (p = 0.01). They both overestimate the number of cigarettes needed daily before smoking becomes toxic (p < 0.01) and are less active physically than nonsmokers (p < 0.007). Heavy smokers live in larger communities (p < 0.02) and smokers are younger than non-smokers (p < 0.0001). In the female population, the level of awareness did indeed influence their behaviour towards cancer prevention (p < 0.04). The level of knowledge was higher in both those who consciously attend cancer screening clinics (cervical Papanicolaou smears and faecal tests) and those who are opposed to faecal tests. Younger females are significantly less aware than those who are older (p = 0.0001) and more than two thirds of these would not consider a cervical Papanicolaou smear as a preventative diagnostic tool. Females who conscientiously attend cancer screening clinics and who are very much aware of cancer, are generally older in age (p < 0.05). The occurrence of mammography as a preventative measure is both high and dependent on age. In conclusion, the level of knowledge of cancer cannot explain alone human behaviour towards the prevention of this disease. Other determinants responsible for influencing human behaviour towards cancer prevention will be better understood once the social sciences, epidemiology and anthropology units work together and combine resources to achieve a common goal. PMID- 9752285 TI - [Parcours de femmes. Opinion survey carried out among women treated for gynecologic and breast cancers and their medical care teams]. AB - The opinion survey "Parcours de femmes" initiated in partnership with Bristol Myers Squibb was conducted by la Ligue nationale contre le cancer between November 1993 and May 1995 and by Sofres Institute. The aim of the study was to increase knowledge of experience of feminine cancers, both by patients and by different treatment teams. Two thousand eight hundred and seventy-four women treated for one of these diseases answered anonymously on a form with sixty-eight questions. Eighty-one individual interviews gave the opinion of the general practitioners and specialists concerned, as well as nursing and hospital staff, pharmacists, health leaders in the private or public fields. The results of this opinion survey highlight and confirm needs, deficiencies, and aspirations of everyone involved. The data show the importance of information, explanations and dialogue at every stage of the disease. The data specify the lines of support to be developed: calming distress, stimulating hope and the will to recover. They also indicate a need for psychological support outside the family and the medical environment as well as for treatment teams. The necessity to improve the hospital environment as well as access to maternal aid during and after treatment to make reinsertion easier are also demonstrated. Apart from associations and help services (such as home care), institutionals do not all have the same sensitivity expect to this pathology. Many people consider it has nothing specific in comparison to other pathologies or to other women in difficulties. They reject the idea of implementing specific measures and valorize research and prevention actions. PMID- 9752287 TI - [The French consensus conference on cancer of the colon: simple and precise recommendations]. PMID- 9752286 TI - [Congress in Saint-Gallen (25-28 February 1998). Adjuvant treatments for operable cancer of the breast: what conclusions?]. PMID- 9752288 TI - [The BRCA2 gene protein, whose disorders predispose to familial cancer of the breast, intervene in DNA repair]. PMID- 9752292 TI - [Acute promyelocytic leukemia, histone deacetylase, and response to retinoids]. AB - Acute promyelocytic leukemia (APL) originate from chromosomal translocations generating two types of fusion proteins both involving the retinoic acid receptor alpha (RAR alpha) and either the gene PML (t(15;17)) or PLZF (t(11;17)). Recent publications cast a new light on the detailed molecular mechanism underlying the oncogenic activity of these fusion proteins which block myeloid terminal differentiation by recruiting histone deacetylases to the promoters of target genes through co-repressor proteins. They also explain the different responses to treatment by all-trans retinoic acid (ATRA) of these two variants which are otherwise clinically indistinguishable. PMID- 9752291 TI - [Results of the NSABP clinical trial for the prevention of cancer of the breast]. PMID- 9752293 TI - [Alternative protein p19ARF: a genuine tumor suppressor gene]. AB - The p16INK4a gene located in the MTS1 (multiple tumor suppressor 1) locus encodes two proteins, p16INK4a and p19ARF, which are structurally unrelated but functionally similar as both inhibit cell cycle progression. In this review, we report and comment new data obtained from mice knockouted for p19ARF without interfering with the expression of p16INK4a. This results in the rapid occurrence of a spectrum of tumors. These data establish that p19ARF is a genuine tumor suppressor protein and raise the questions of the relationship between the two proteins and their respective importance in malignancies. PMID- 9752294 TI - [Interferon and retinoic acid in the treatment of human cancer: mechanisms of action]. AB - Retinoic acid (RA) and interferons (IFN) are negative regulators of cell proliferation. A number of clinical trials were thus carried out in cancer therapy with RA and/or IFN. In vitro and in vivo, their combination leads to a more potent cell growth inhibition. Moreover, RA and IFN act cooperatively to increase the expression of many IFN-stimulated genes, leading also to a higher cell differentiation and inhibition of viral replication. However, the molecular mechanisms by which RA and IFN potentiate each other are not fully understood. The cooperative effects by RA and IFN are mediated through multiple pathways. RA causes the induction and secretion of IFN alpha. RA also stimulates the IFN regulatory factor gene expression (IRF1 and p48). Additional mechanisms could be involved as RA increases the level of signal transducing activators of transcription (Stat) proteins, and thus enhances the IFN-induced Stat activation. PMID- 9752296 TI - [Sphincter conservation and cancer of the lower rectum: argument for a multicenter prospective study for conservation of the sphincter after irradiation]. AB - Rectal adenocarcinoma located less than 6 cm from the pectineate line raise the issue of sphincter preservation. Preoperative treatment response can modify the initial indication of surgery. With good response, conservative surgery can be performed with a distal margin less than 2 cm. Conversely a lack of response requires an abdominoperineal resection. Removal technique must emphasize a total mesorectal excision and a prevention of urosexual complications. Intersphincteric resection allows to increase distal margin without continence dysfunction. Regional lymphadenectomy doesn't increase regional control. Digestive tract reconstruction combines coloanal anastomosis and J colonic pouch whenever anatomical conditions are favorable. Long-term follow up should validate this therapeutic approach. PMID- 9752295 TI - [Microinvasive carcinoma uterine cervix. Which approach in 1998?]. AB - Numerous definitions of microinvasive carcinoma (MIC) have been proposed. The SGO takes into account the depth of stromal invasion and presence of capillary like space involvement (LVI). The Figo uses the lesion width and describes different substages according to the depth of stromal invasion. Two major prognostic factors can be identified in the literature: the depth of invasion and the presence of LVI. The lesion volume is probably more accurate than the depth of stromal invasion but cannot be measured in routine. Taking into account that a classification must be a guide for the evaluation of prognosis and treatment, the SGO definition seems more reliable. Pelvic lymph node metastasis rate and recurrence increase with these two factors. MIC with stromal invasion under 3 mm and without LVI have a little risk of parametrial and nodal involvement: with a high rate of survival. Conversely, MIC with invasion over 3.1 mm depth or LVI have a greater risk of spread beyond the cervix (1% versus 7.7%) and many authors now consider them as true invasive cancers. For lesion invading the stroma within 3 mm, the treatment can be limited to a standard hysterectomy with good results. Some authors have proposed more conservative therapy as conization. This procedure is interesting for young women willing to preserve their anatomy, fertility and sexual function. In selected cases, short term results are similar to those of hysterectomy but there is a lack of controlled studies with long term follow-up. Lesions over 3.1 mm with LVI should be treated as true invasive cancers. Intermediate cases should have a conservative therapy associated with a laparoscopic lymphadenectomy. PMID- 9752297 TI - [Internet: importance in cancerology and practical connection sites]. AB - There has been a considerable increase in World Wide Websites on the Internet on medicine and oncology. The clinician has access to an abundance of useful documentary and graphic information. The challenge is firstly to find a relevant site and then to evaluate the quality of the material available on that site. In future, there may well be a service on the network which reviews sites. The aim of this paper is to give practical guidance to connecting on the Internet as well as a list of the best sites for clinicians, researchers, students and oncologists. PMID- 9752298 TI - [Standards, Options, and Recommendations for using erythropoietin in cancerology]. AB - The "Standards, Options and Recommendations" (SOR), started in 1993, are a collaborative project between the Federation of the French Cancer Centres (FNCLCC), the 20 French Cancer Centres and specialists from French Public Universities, General Hospitals and Private Clinics. The main objective is the development of clinical practice guidelines to improve the quality of health care and outcomes for cancer patients. The methodology is based on literature review and critical appraisal by a multidisciplinary experts group, with feedback from specialists in cancer care delivery. OBJECTIVES: To develop a clinical practice guideline with definitions of Standards, Options and Recommendations for the use of recombinant human erythropoietin (rHuEPO) in oncology. METHODS: Data have been identified by literature search using Medline (up to march 1996) and Current Contents (up to october 1996) and personal references lists. The main end points considered were hemoglobin level, haematocrit, quality of life, transfusion requirements, incidence and length of hospital stays, efficacy of cancer treatment, safety and costs. Once the guideline was defined, the document was submitted to 39 reviewers for peer review, and to the medical committees of the 20 French Cancer Centres for review and agreement. RESULTS: The key recommendations are: 1) the use of recombinant human erythropoietin in oncology is validated for chemotherapy-induced anemia when the chemotherapeutic regimen contains platinum. In other cases, we recommend to suggest patients participating in prospective clinical trials; 2) for chemotherapy (platinum based)-induced anemia, the benefits/risks ratio for anemia therapy (i.e. transfusion or erythropoietin therapy) must be analysed for each individual patient; 3) we recommend participation in studies to identify predictive factors for non response to erythropoietin therapy to select non-responding patients; 4) to investigate the clinical benefit of erythropoietin therapy for anemia due to intensive cytotoxic chemotherapy and radiation therapy, we recommend to suggest patients participating in large multicentre phase III trials; 5) at the present time, there is insufficient evidence to recommend the use of erythropoietin therapy in children. PMID- 9752299 TI - [Long-term prognostic role of steroid receptors in cancer of the breast]. AB - We screened for the prognostic value of estrogen receptor (ER) and progesterone receptor (PR) through a multicentric study of 2,257 operable breast cancer patients who did not received adjuvant therapy. Three hundred and seven local regional recurrences, 105 metachronous contralateral breast cancer, 589 metastases and 537 deaths from cancer had been diagnosed with a median follow-up of 8.5 years. A total of 69% of the tumors were ER positive and 54% PR positive. For statistical analysis, 1,665 patients were studied because of complete clinical and biological data. In univariate analysis, ER and PR status were of prognostic value for the metastases-free interval (MFI) and the overall survival (OS). In multivariate analysis (Cox proportional hazard model), only the ER status showed a significant difference between positive and negative groups regarding the MFI and OS. By using Cox regression model with time-dependent covariates, we show that the predictive value of ER status of the primary tumor decreases by approximately 20% per year, losing its significance after 8 years of follow-up. These results show that ER and PR status have a relatively limited predictive value and their major interest remain in the domain of therapeutic decision. PMID- 9752301 TI - [Evidence-based medicine (EBM)...an evidence?]. PMID- 9752300 TI - [Morphine therapy: evaluation of patient information]. AB - Pain is frequent in the course of cancer and can have negative consequences on patients quality of life. The great majority of patients can be helped by simple treatments. The prescription of morphine (M) must be preceded by some explanations. In order to verify the reality of these explanations, a study was done in a 230 comprehensive beds Cancer Centre, with 129 patients, randomized between all the patients hospitalized. In an open questionnaire, different aspects were studied. One hundred answers were studied: 63 patients did not have M at any time: group M-; 37 patients had M (19) or have had M (18): group M+; 97% of the patients in group M+ thought that M decreased pain, for 85% patients in group M-. Morphine treatment was effective in 92% of patients M+, and not very effective in 5%; 67% of patients M- thought that M is efficient but 17% did not know; 76% of patients M+ did not worry about M; 13% worried et 11% did not know. For patients M- only 48% did not worry; 41% worried and 11% did not know. The side effect known by the patients, and spontaneously quoted were constipation (12 patients in M+ group). For 14 patients there was no problem and 2 did not know. In M- group 49 patients did not know. Only 8% of M+ group were afraid by addiction, but 44% in group M-; 97% patients M+ said that they could stop without problem against only 38% of the patients in M- group. For 16% of M+ group, the use of M had a bad signification about their disease, but 52% of M- group thought that if morphine was used in their cases, the meaning would be very bad. The differences between the two groups allow to think that the prescription of M in this study is explained, and that the patients receiving M are rather well informed. Patients with cancer but without M do not have good information and their knowledge is similar than general population. The use of recommended therapy and explanations allowed patients with M therapy to be comfortable with this prescription. PMID- 9752302 TI - [The alpha/beta connection]. PMID- 9752303 TI - [Cancer of the endometrium. Epidemiology, anatomopathology, screening, diagnosis, evolution, prognosis, treatment principles]. PMID- 9752304 TI - [Targeting of doxorubicin to the tumor vascular system]. PMID- 9752309 TI - [Fluorescent in situ hybridization in cancerology]. PMID- 9752308 TI - [The gene of Gorlin's syndrome (basocellular nevomatosis) and the revival of the developmental genes in human carcinogenesis]. AB - Interaction between developmental biology and human pathology has brought new insight on basal cell carcinoma oncogenesis. Alterations of the Patched gene appear to be responsible for the Gorlin syndrome and to be detected in 50% of the sporadic basal cell carcinomas. This relation has recently been extended to all the components of the Hedgehog signalling pathway. PMID- 9752310 TI - [Secondary non-Hodgkin's lymphomas]. AB - Developing a secondary non Hodgkin's lymphoma (NHL) more than 12 months after treatment for first cancer, is uncommon. Secondary NHL occurs sometimes after chronic lymphatic leukemia or Hodgkin's disease. The 20 years cumulative incidence rate is 3-5% after Hodgkin's disease. Secondary NHL seems less frequent after any childhood cancer. Pertinent features of secondary NHL are a high percentage of patients with extranodal tumor sites (brain, digestive tractus), high grade histological subtype, phenotype B and advanced stage. But for identical histological type and stage, the prognosis of secondary NHL seems the same than de novo NHL ones. The incidence of secondary NHL is associated to individual parameters (age), first cancer (chronic lympho proliferative syndrome, Hodgkin's disease) and previous chemo- and/or radiotherapy. Immunodeficiency is probably the most important cofactor for the subsequent development of secondary NHL. However, secondary NHL differs from NHL of immunosuppressed patients (HIV+, posttransplant) because brain lymphoma are less frequent and immunodeficiency disorders unknown. Secondary NHL are also different of therapy-associated leukemias in the late occurrence after the first treatment and in the questionable role of cytotoxic agents given for the treatment of the first cancer. PMID- 9752311 TI - [Dose-effects and chemotherapy dose intensity of aggressive non-Hodgkin's lymphomas in the adult]. AB - The CHOP regimen (cyclophosphamide vincristine, adriamycin, prednisone) is considered since twenty years as the standard treatment of disseminated aggressive non-Hodgkin's lymphomas and cures approximately 30% of patients. More recently, intensive chemo(radio)therapy followed by rescue with bone marrow or blood hematopoietic stem cells has become the standard treatment of chemosensitive relapses of aggressive non-Hodgkin's lymphomas. Consequently, two questions with practical applications have arisen and are discussed in this review: Is it justified to increase the dose intensity of chemotherapy, using either single treatment intensification with stem-cell rescue, or sequential intensified chemotherapies, especially in cases with adverse prognostic factors? On the contrary, is a decrease in treatment intensity compared to CHOP chemotherapy, especially in elderly patients, harmful to the patient? PMID- 9752312 TI - [Oncogenetics: a new activity between research and medicine]. AB - The on-going development of predictive tests for common cancers--as breast and colorectal cancers--is potentially introducing fundamental changes in medical practices, and, therefore, changes in the role and meaning of medicine in society. In this context, analysing the conditions of emergence and development of cancergenetics activities, and their potential implications, appears as an issue of particular interest for sociology. The study conducted at Inserm unit 379 in Marseille, in collaboration with physicians from institut Paoli-Calmettes, led us, firstly, to identify a set of factors as decisive for the future of these activities. In addition, this analysis highlights the diversity of the conditions of introduction of genetic tests, depending on the pathologies and the context of existing clinical practices. Considering these elements, we argue for the setting of adequate modalities of regulation so as to permit a managed and responsible introduction of genetic testing in medicine and society. PMID- 9752313 TI - [Oncogenetics: how far, and what for?]. AB - Oncogenetics is a new discipline expanding very rapidly and depending on advances in understanding genes associated with inherited susceptibility to common adult malignancies. This discipline is still between a research field and a clinical service. This is very specific regarding medical management and services related to cancer genetic testing, directions for use genetic testing. It is the emergence of a so-called "predictive" medicine and in general of problems surrounding genetic testing for adult-onset disorders susceptibility. In the past years significant and unprecedented media fanfare informed both medical professionals and the general public. The opinions have moved from enthusiasm to skepticism but now a fair approach should be obtained. A good example of current controversies is breast cancer susceptibility. At the present time the oncogenetics debate "yes ... but on condition that ..." replace the previous one "in favour or against". The remained relevant topics are "how far" and "what for". Specific guidelines are already published or are on-going development. All of them point out a critical need for assessment (long-term outcome studies, multidisciplinary team approach for medical management, patient oriented research to analyse the psychological and social impact of genetic testing). PMID- 9752314 TI - [Ethics and oncogenetics: how to resolve the contradictions?]. AB - Ethical guidelines are necessary to help practitioners to deal with matters about which they are uncertainty and to solve problems where conflicting interests are at stake. To avoid doing harm is the first criterion to be adopted, in line with primum non nocere, as doing good is not enough (saving lives is not sufficient and one also has to consider whether a loss of dignity or suffering will not make the benefit worthless). Consultees must come to consultations freely and intentionally and must have control over the choices subsequently made. But others interests may be involved and ethical misconducts can arise in situations such as the following: family pressure about DNA testing, medical pressure to help other patients of for scientific purposes (sometimes merely to obtain data for publication). Geneticians must look at the reality underlying each situation, although complete autonomy and neutrality may be impossible to achieve. Individuals' rights are by now recognised, but the pursuit of autonomy at all costs has led to the development of a kind of self service medicine. One extreme situation of this kind was the request for prophylactic mastectomy made by a mutation free woman from a high risk family. Since this woman had seen her mothers and sister(s) die from breast cancer, her appraisal of her own risk may not fitted the neutrally calculated cumulative lifetime risk, which is about 9% (the so-called sporadic risk). Practitioners perceive contradictory elements during their consultations, and they ask for help. They cannot solve this kind of situation by themselves even by consulting the available medical, statistical and scientific knowledge. Other specialists (humanities) have to be consulted and asked to avoid wrong counselling. PMID- 9752315 TI - [Managing a long term illness: the point of view of the sociologist]. AB - This paper presents a sociological perspective of the experience of the cancer patient. Cancer is considered in the wider context of chronic and/or long-term illness which are typical of modern societies. The sociological concepts of "biographical disruption", "illness trajectory", "endless work and care" of the patient and his family are useful for analysing the experience of illness, and doctor-patient relationships. Moreover, they might illuminate some aspects of the individual and collective impact of modern oncogenetic technologies. PMID- 9752316 TI - [Chromosome translocations and leukemias induced by inhibitors of topoisomerase II anticarcinogenic drugs]. AB - The treatment of cancer with alkylating drugs or topoisomerase II inhibitors can be responsible for the development of myelodysplastic syndromes and acute myelogenous leukemia. Alkylating agents such as melphalan and cisplatinum mainly produce damages at chromosomes 5 and 7 whereas topoisomerase II inhibitors induced lesions essentially affect chromosomes 11 and 21. Rearrangements of the MLL gene at band 11q23 are frequently observed in human de novo myeloid and lymphoid leukemia as well as in leukemia or myelodysplasia secondary to therapy with drugs targetting topoisomerase II such as the epipodophyllotoxins. A relationship between the treatment with etoposide on teniposide and the development of translocations of the MLL gene has been clearly evidenced. The potential molecular basis of the chromosomal rearrangements implicating topoisomerase II and its inhibitors are discussed. The chemical structure of the inhibitors, their mechanism of action and the genes targetted by these drugs are presented. DNA cleavages induced directly by topoisomerase II inhibitors or by the drug induced apoptotic cellular response are responsible for nonrandom chromosomal aberrations and contribute to leukemogenesis. PMID- 9752318 TI - [Importance of micronucleus tests in cultured binuclear T lymphocytes for the detection of genotoxic events in cancer patients]. AB - The cytokinesis-blocked micronucleus assay (CBMN) is a short-term mutagenesis test which offers an easier and less tedious alternative to metaphase chromosome analysis, with the advantage that exposure to both clastogens and aneugens may be detected. The CBMN assay has been used in evaluating the genotoxic consequences of exposures to environmental and occupational mutagens and carcinogens. Micronucleated cell rates (MN cell rates) were assessed in cytokinesis-blocked lymphocytes of 70 male and female cancer patients prior to any anticancer treatment. The study of interindividual variation factors showed that only age significantly affect MN cell rate, whereas sex, tobacco, alcohol, imaging techniques and tumour stage had no significant effect. The comparison of micronucleated cell rates in 198 healthy subjects and 70 cancer patients matched for age and sex showed a statistically significant difference. Spontaneous elevated MN cell rates of cancer patients refer to previous exposition of genotoxic or mutagenic environmental agents. Moreover, the MN cell rates in cancer patients most probably refers to various cellular lesions and genetic damages. PMID- 9752319 TI - [Integration of economic criteria in the recommendations for clinical practice in cancerology]. AB - Clinical practice guidelines have been defined as "systematically developed statements to assist practitioners and patients in their decisions about appropriate health care for specific clinical circumstances". Their objectives are to improve the quality of health care and to optimise the use of limited health care resources. However reduction of unnecessary costs of delivered health care is proceed most often in an implicit way by identifying inappropriate health care strategies. The increase of health care costs needs to look at this issue in a more explicit way and to consider costs in the guideline development process. The key objective of our study is to analyse the methodological aspects of dealing with cost issues in the guideline development process. The integration of cost issues is in fact limited by two major problems: first, the lack of economic evaluation for many strategies in the scientific literature and second, the lack of generalizability of the published results to temporally and/or geographically different settings. These difficulties are likely to result in the need for local cost evaluation (for a given setting), and though to make the guideline development process much more complex. Further methodological research is important to define the role of economic evaluation in clinical practice guidelines and to enable the integration of cost issues into the guideline development process. They should go closely together with international standardisation of the methodology for designing, conducting and reporting economic evaluation. PMID- 9752317 TI - [Biological markers for the prognosis of neuroblastoma: proposal of a method of analysis]. AB - The prognosis of pediatric neuroblastoma depends both on clinical presentation and on certain cellular and molecular characteristics. At the present time, two hypotheses can be drawn to explain both clinical and biological heterogeneity. In the first hypothesis, neuroblastoma progresses from early to late clinical stages through a classical multistep process linked to an accumulation of molecular abnormalities. In the second hypothesis, neuroblastoma represents an heterogeneous group of unrelated diseases, where most of stages I and II or stage IVS neuroblastomas can rather be considered as benign tumors, and stage IV neuroblastoma as a true malignant proliferation. To ascertain relevant biological factors for the prognosis of the disease, it is uppermost important that all investigators agree on biological criteria for analysis when neuroblastoma tissue is available in screened and unscreened populations. This paper reviews the biological tools available for prognosis in neuroblastoma, the priority for analysis of biological markers according to reliability, feasibility, and reproducibility of analysis procedure, and the conditions of tissue storage for further analysis of these biological markers. The standardized biological evaluation of neuroblastoma will allow, first, to collect sufficient data for multivariate analysis of prognostic factors and, second, to better define the putative links between various forms of the disease. PMID- 9752321 TI - [Telomeres, telomerase: the year 1998 has begun well]. PMID- 9752320 TI - [Electrophoresis in denaturing gradient gel (DGGE)]. PMID- 9752323 TI - [Bone density and risk of cancer of the breast in menopausal women]. PMID- 9752324 TI - [Telomerase and cancer: correlation or causality?]. PMID- 9752327 TI - [1997, a good year for telomerase?]. AB - Telomerase is a ribonucleoprotein enzyme specialized in telomere elongation which may be involved in the control of cell proliferation, and consequently could play a role in oncogenesis, 1997 was a very exciting year for telomerase understanding. First, the sequence of the human telomerase catalytic subunit revealed strong homologies with known reverse transcriptases. Second, the production of viable mice deleted for the RNA moiety of the enzyme showed that telomerase is essential to maintain telomere length in germ cells but is not required for tumour formation. PMID- 9752325 TI - [in Vitro reconstitution of human telomerase activity]. PMID- 9752328 TI - [Management of nonresectable non-small cell lung cancer. Recommendations of the American Society of Clinical Oncology (ASCO)]. PMID- 9752330 TI - [BCG therapy of superficial tumors of the bladder]. AB - Superficial bladder tumor defined as pTa, pT1 and pTis stage is one of the most common cancer disease in the world. Intravesical BCGtherapy represents the best treatment when combined with transurethral resection to prevent tumor recurrence or progression and to lengthen the interval between recurrence. The toxicity is quite important but can be reduced by using the good procedure. Local or general immunostimulation is likely to be the exact mechanism of action but more data are needed. PMID- 9752329 TI - [Fas, fas ligand, immune tolerance, and cancer: implications in cancer of the colon]. AB - Fas and Fas ligand (FasL) are implicated in programmed cell death or apoptosis. In the immunological field, they are particularly important in auto-immunity, graft rejection and anti-tumoral response. Fas ligand expression on thymocytes, activated T lymphocytes, and in sites of immune privilege, suggests the importance of Fas/FasL interactions in negative control of the immune response. The recent description of FasL expression by tumoral cells, represents a new mechanism of immune escape for different cancer, and has been well studied in colon adenocarcinoma. PMID- 9752331 TI - [Evaluation of the quality of life in clinical research in cancerology]. AB - This article clarifies the methodological aspects of quality of life evaluation and its use in cancer research. The quality of life of an individual is a complex, multidimensional, subjective concept, the perception of which is expressed optimally by individual his/herself. The tools most often used to measure the quality of life in clinical research are based on psychometry. They take at least three dimensions into account: physical, psychological and social. They must be valid, reliable and sensitive. These qualities must be verified again when translated into another language. A review of the main tools is provided. The place to be assigned to quality of life evaluation in clinical research is discussed. Routine evaluation is to be excluded because the quality of life is a subjective phenomenon that is tricky to measure and this evaluation increases the temporal and financial costs of the study, without forgetting the intrusive aspect of this action. On the other hand, as long as it contributes substantially to decision-making regarding the choice of curative or palliative treatments, or interventions necessary for the rehabilitation of the patient, it is an assessment criterion that must be included in the study. PMID- 9752332 TI - [Standards, Options, and Recommendations: a space in Bulletin du Cancer to facilitate their dissemination]. PMID- 9752333 TI - [Standards, Options, and Recommendations for the surveillance after treatment in cancer of the colon. The SOR Working Group "Cancer du Colon" de la Federation Nationale des Centres de Lutte Contre le Cancer]. PMID- 9752334 TI - [Modelling of intraperitoneal chemohyperthermia: experimental study and identification of certain thermal aspects]. AB - Intraperitoneal chemohyperthermia is more and more considered as an interesting therapeutic option in cases of some abdominal cancers, particularly of digestive origin. However, many technical aspects of this treatment remain far from being mastered, particularly the homogeneous dispatch of temperatures within the abdomen cavity. This work consists, first of all, in an experimental study, which is being carried out on a physical "prototype" of the abdomen, on which different hot fluid flows and injection conditions (configurations) are investigated. The results of this experimental study are prospected in two ways. First, an a priori thermal model is proposed, based on physical equations (heat transfer, etc.). Then a "black box" model is identified from the measured temperatures evolutions, so as to obtain a model of the "system" behaviour. Finally, the two modelling approaches are being compared, and the results converge to a simple expression of a few parameters, either physical or identified. These modelling results have helped to optimize the injection circuit and its running parameters while applied to the human treatment. PMID- 9752335 TI - [Prosthetic breast reconstruction and radiotherapy: analysis of 67 cases)]. AB - Authors inquire into repercussions of radiotherapy (RT) on prosthetic mammary reconstruction (PMR) by using the retrospective analysis of 67 cases, 59 of which were performed in irradiated areas. The particular aspect of irradiated prosthesis is evoked regarding 8 cases, and with the support of a literature review. Surgical complications and late results are evaluated comparatively to 339 PMR among non irradiated patients. According to available data in our series, harmfulness of RT not appeared statistically demonstrated. From both their experience and the literature, authors clarify indications of PMR following RT. They conclude to the necessity of a high quality RT, and optimal adaptation of surgical technique to tissular conditions. PMID- 9752336 TI - [Choice and disclosure of preferences, towards sharing the therapeutic decision in cancerology: from economic theory to medical practice]. AB - Today, as it is often difficult to demonstrate the superiority of a new molecule or a therapeutic strategy in term of plain efficacy on disease, the incitement is strong to provide some complementary argument of assessment, we are assisting to the emergence of a new concept: shared therapeutic decision making. Is the application of this concept--with make the paternalistic model questionnable- adapted to all cases? What are the different levels of participation that could be envisaged? Are there favourable methods for this participations? This shared decision making--direct (patients' choice between treatment options) or indirect (integration of elicited preferences in the decision process)--if it has to be efficient, must surround with care: to define its application limitation, to protect itself of manipulation. It shall require to consider information transmission difficulties, to establish some elicitation preference method. Some technical, such as time trade off, standard gamble or willingness to pay, supported by economic theory of expected utility, permit to help eliciting patients' preferences and to structure the therapeutic choice. Some empirical study of preference elicitation shall permit to get clear the complexity of trade off between the different choice element that could enter in the acceptability of the treatment for patients. PMID- 9752337 TI - The development of a French version of a questionnaire on the quality of life in cancerology (Functional Living Index-Cancer: FLIC). AB - The quality of life has become an indispensable element in the evaluation of treatments, especially as clinical trials are generally conducted in more than one country. In cancerology, numerous questionnaires have been developed and validated, but most of them are in English. This article presents the stages of translation and validation of the French version of the FLIC (Functional Living Index-Cancer). It applies to a sample of 200 patients, 47% of whom have breast cancer. The acceptability of this questionnaire is good (92.2%). The main psychometric criteria were verified. The Cronbach alpha coefficient, equal to 0.90, shows good reliability. A factor analysis was conducted to examine construct validity; it revealed a 5-factor solution accounting for 62% of the variance. These 5 factors are associated with specific domains; physical, psychological and social functioning, current well-being and disease symptoms. The global scores of the FLIC correlate significantly with those of the different scales of the EORTC QLQ-C30. Moreover, clinical relevance is ensured by its correlation with certain clinical variables (WHO index, location of tumour). The French version of the FLIC can be used to measure the quality of life of patients afflicted with cancer, but further studies are necessary to confirm its reliability. PMID- 9752338 TI - [Melanoma: current controversies and future perspectives]. PMID- 9752339 TI - [Update on molecular mechanisms of carcinogenesis]. PMID- 9752340 TI - [Update on genetic markers of cancer]. PMID- 9752341 TI - [Update on tumor immunology]. PMID- 9752342 TI - [Update on the study of the breast]. PMID- 9752343 TI - [Update on gynecology]. PMID- 9752344 TI - [Update on gastroenterology]. PMID- 9752345 TI - [Update on malignant hematologic diseases]. PMID- 9752346 TI - [Update on lymphomas: 1997, a profitable year]. PMID- 9752347 TI - [Update on cancerology of the upper aerodigestive tract]. PMID- 9752348 TI - [Update on pulmonary cancerology. What is new in 1997]. PMID- 9752349 TI - [Update on neuro-oncology]. PMID- 9752350 TI - [Update on pediatric oncology]. PMID- 9752351 TI - [Soft-tissue sarcomas in the adult: news and trends]. PMID- 9752352 TI - [Update in epidemiology]. PMID- 9752354 TI - [Update on tumor biological markers: reality and perspectives in 1998]. PMID- 9752353 TI - [Update on medical imaging in cancerology]. PMID- 9752355 TI - [Update on cancer surgery]. PMID- 9752356 TI - [Update on radiotherapy]. PMID- 9752357 TI - [Update on antitumor pharmacology: reality and perspectives in 1998]. PMID- 9752358 TI - [Update on cytokines and growth factors]. PMID- 9752360 TI - [Update on the ethics of clinical research]. PMID- 9752359 TI - [Update on palliative treatments]. PMID- 9752361 TI - Lumbar supports to prevent low back pain. PMID- 9752362 TI - Beta-blockers for hypertension in the elderly. PMID- 9752364 TI - Short-course antibiotics for acute otitis media. PMID- 9752363 TI - Early administration of ACE inhibitors in AMI. PMID- 9752365 TI - Conservative management of non-Q-wave myocardial infarction. PMID- 9752367 TI - Exercise therapy for dizziness and vertigo. PMID- 9752366 TI - Glucocorticoids and antibiotics in preterm PROM. PMID- 9752369 TI - Advancing information mastery in family practice. PMID- 9752368 TI - Zinc lozenges for treating the common cold in children. PMID- 9752370 TI - Should low-molecular-weight heparins replace unfractionated heparin as the agent of choice for adults with deep venous thrombosis? AB - BACKGROUND: Several low-molecular-weight heparins (LMWHs) are now approved for use in the United States for the prophylaxis of venous thromboembolism. They are used in Europe for the treatment of deep venous thrombosis (DVT) and pulmonary embolism. This review examines the evidence addressing the question "Should LMWHs replace unfractionated heparin (UFH) in the treatment of adults with DVT?" METHODS: We performed a MEDLINE search using the key words "low-molecular-weight heparin" from the years 1990 to 1998, and the results were assessed using the JAMA Users' Guides to the Medical Literature system. RESULTS: Low-molecular weight heparins are at least as safe and effective as unfractionated heparin in the treatment of patients with DVT. They are probably more effective and safer. They are more convenient to use and are associated with lower overall costs. CONCLUSIONS: Based on efficacy, safety, convenience, and cost, LMWHs are clearly superior to UFH in the treatment of DVT in primary care. Studies that confirm an expected improvement in patient-oriented outcomes (e.g., mortality and quality of life) need to be done. PMID- 9752371 TI - A clinical decision analysis of cryotherapy compared with expectant management for cervical dysplasia. AB - BACKGROUND: Screening for precursors of cervical cancer with colposcopic examination for women with abnormal Papanicolaou (Pap) smears identifies those with cervical dysplasia. Though the majority of mild dysplasias (CIN I) will regress, many are treated with cryotherapy. METHODS: We used decision analysis to compare immediate cryotherapy with expectant management (following with another Pap smear or colposcopy, with treatment reserved for progression or a duration of 2 years). The decision tree included the possibility of more invasive surgical procedures if the cryotherapy was ineffective or if the dysplasia progressed in extent of cervical involvement or in grade. Probabilities were derived from literature review and expert judgment. The analysis considered the disutility of the follow-up examinations, cryotherapy, and the more invasive procedures, using expert assessment. RESULTS: Using the baseline assumptions, expectant management led to a better outcome for most patients (57%), who recover with no procedure. However, more patients treated with expectant management required surgical procedures (loop electrosurgical excisional procedure, conization, or, rarely, hysterectomy) than did those treated with immediate cryotherapy. In the expected disutility analysis, expectant management was better than immediate cryotherapy. Sensitivity analysis showed that three factors had the potential to change the recommendation of the analysis: (1) the probability the dysplasia will regress, (2) the disutility of the process of expectant management, and (3) the disutility of invasive procedures compared with cryotherapy. CONCLUSIONS: The analysis indicated that expectant management is preferable to immediate cryotherapy for women with histologically proven mild cervical dysplasia. However, this conclusion depended on assumptions about three factors for which there is insufficient evidence in the literature. More research is needed. PMID- 9752372 TI - Patient and visit characteristics associated with opportunistic preventive services delivery. AB - BACKGROUND: This study's purpose was to identify patient and visit characteristics associated with the use of illness visits as opportunities for the delivery of preventive services and to determine if time is allocated differently during illness visits that make use of these opportunities. METHODS: Research nurses directly observed the delivery of preventive services during consecutive patient visits on 2 separate days in the offices of 138 family physicians. Data on patient eligibility for preventive services were collected by medical record review. Time use during patient visits was categorized using the Davis Observation Code (DOC). Patient characteristics, visit characteristics, and time use were compared during illness visits in which at least one service recommended by the US Preventive Services Task Force was delivered to eligible patients, compared with illness visits during which no recommended preventive services were delivered. RESULTS: Preventive services were delivered during 32% of 3547 illness visits. Adults, overweight patients, those who smoke or drink alcohol, new patients, and patients with fewer visits in the past year were more likely to receive preventive services. Patient request was also associated with increased delivery of preventive services. The presence of another family member, visits for an acute illness, and the prescription of a drug were associated with a decreased likelihood of a patient's receiving preventive services. When preventive services were delivered during illness visits, less time was spent on chatting, procedures, and physical examination, and more time was spent on history-taking. CONCLUSIONS: Family physicians take greater advantage of opportunities for the delivery of preventive services during the illness visits of high-risk patients. The results of our study suggest strategies that could be used to expand the opportunistic delivery of preventive services to other patients and types of visits. PMID- 9752373 TI - Changes in functional health status during normal pregnancy. AB - BACKGROUND: Pregnancy is a period of physical and emotional stress that can have a significant impact on the well-being of an expectant mother. To explore the extent to which normal pregnancy affects the life of a mother, we sought to assess changes in a standard quality-of-life measure throughout normal pregnancy. METHODS: We performed serial assessments of health-related functional status in a cohort of 125 healthy women who obtained care from a residency-training clinic during pregnancy. Before each antenatal visit, women completed the Medical Outcomes Study Short-Form 36 (SF-36) along with a questionnaire to assess any complications with their pregnancy. Scores for each of the SF-36 subscales were calculated and examined to determine the influence of gestational age and socioeconomic factors on quality of life during pregnancy. RESULTS: Of the eight subscales of the SF-36, three of the four associated with physical health status changed significantly with gestational age. Physical functioning (P < .001), role limitation due to physical problems (P < .001), and pain scales (P < .001) all decreased as pregnancy progressed. These scales showed linear decreases during the first two trimesters, with relative flattening in the third trimester. Sociodemographic factors such as employment, level of income, and presence of a spouse or support partner had only a small influence on functional status in pregnancy. CONCLUSIONS: Health-related functional status during pregnancy changes only for physical measures of health. Sociodemographic factors appear to have only limited influence on health-related functional status during pregnancy. PMID- 9752374 TI - Linking primary care performance to outcomes of care. AB - BACKGROUND: Substantial research links many of the defining characteristics of primary care to important outcomes; yet little is known about the relative importance of each characteristic, and several characteristics have not been examined. These analyses evaluate the relationship between seven defining elements of primary care (accessibility, continuity, comprehensiveness, integration, clinical interaction, interpersonal treatment, and trust) and three outcomes (adherence to physician's advice, patient satisfaction, and improved health status). METHODS: Data were derived from a cross-sectional observational study of adults employed by the Commonwealth of Massachusetts (N = 7204). All patients completed a validated questionnaire, the Primary Care Assessment Survey. Regression methods were used to examine the association between each primary care characteristic (11 summary scales measuring 7 elements of care) and each outcome. RESULTS: Physicians' comprehensive ("whole person") knowledge of patients and patients' trust in their physician were the variables most strongly associated with adherence, and trust was the variable most strongly associated with patients' satisfaction with their physician. With other factors equal, adherence rates were 2.6 times higher among patients with whole-person knowledge scores in the 95th percentile compared with the 5th percentile (44.0% adherence vs 16.8% adherence, P < .001). The likelihood of complete satisfaction was 87.5% for those with 95th percentile trust scores compared with 0.4% for patients with 5th percentile trust scores (P < .001). The leading correlates of self-reported health improvements were integration of care, thoroughness of physical examinations, communication, comprehensive knowledge of patients, and trust (P < .001). CONCLUSIONS: Patients' trust in their physician and physicians' knowledge of patients are leading correlates of three important outcomes of care. The results are noteworthy in the context of pervasive changes in our nation's health care system that are widely viewed as threatening to the quality of physician patient relationships. PMID- 9752375 TI - Rural childhood immunization. Rates and demographic characteristics. AB - BACKGROUND: Childhood immunization rates are suboptimal, especially in high-risk populations. Rural residents could constitute a population at high risk for childhood underimmunization; little is known about demographic factors associated with childhood underimmunization. This study compared the immunization rates of urban and rural 2-year-olds and examined the association between demographic factors and underimmunization for rural 2-year-olds. METHODS: We analyzed two nationally representative surveys: the 1991 National Maternal and Infant Health Survey (NMIHS) and the 1993 National Health Interview Survey (NHIS). The study population consisted of children in non-metropolitan statistical areas who were 24 to 36 months of age in the NMIHS and 19 months to 5 years of age in the NHIS. The NMIHS sample contained 4425 children (966 in rural areas) and the NHIS sample contained 2505 children (566 in rural areas). RESULTS: There were no significant differences in immunization rates between rural and urban children. In urban areas, immunization rates were 63.3% (NMIHS) and 65.5% (NHIS) compared with 63.0% (NMIHS) and 67.8% (NHIS) in rural areas. Low income, low family education, nonwhite race, unemployment, and being a female child were associated with underimmunization in one or both data sets. These relationships were not modified by residence in a universal purchase state, where the state purchases and distributes vaccine for all children to reduce the cost and thereby improve access to immunization services. CONCLUSIONS: Approximately one third of children in urban and rural areas were underimmunized. The demographic characteristics of underimmunized children were similar in urban and rural areas; however, the special characteristics of rural areas may require that interventions be tailored to rural needs. PMID- 9752376 TI - Genetic diagnosis in adulthood. A case report. AB - While family physicians may readily entertain genetic diagnoses in their pediatric patients, they may fail to consider such diagnoses in their adult patients. We present the case of a man with recurrent leg ulcers who was recognized as hypogonadal and was ultimately given the diagnosis of Klinefelter's syndrome (XXY) at age 47. Although there is no primary treatment for XXY, significant associated conditions, including osteoporosis and testosterone deficiency, can be ameliorated. We review the clinical condition of XXY at various ages and summarize age-specific interventions. We discuss the importance of genetic diagnosis throughout the life span. PMID- 9752377 TI - A foodborne outbreak of cyclosporiasis caused by imported raspberries. AB - BACKGROUND: Cyclospora cayetanensis is a recently recognized parasite that causes prolonged diarrheal illness. Its modes of transmission have not been fully determined, although some investigations before 1996 implicated water. Outbreaks of cyclosporiasis in the United States in 1996 and 1997 are evidence of the increasing incidence of this disease. This report describes an outbreak of cyclosporiasis in persons who attended a luncheon on May 23, 1996, near Charleston, South Carolina. METHODS: In this retrospective cohort study, we interviewed all 64 luncheon attendees and the chef regarding food and beverage exposures. A case of cyclosporiasis was defined as diarrhea (> or = 3 loose stools per day or > or = 2 loose stools per day if using antimotility drugs) after attending the luncheon. We identified sporadic cases of cyclosporiasis and traced the implicated food. RESULTS: Of 64 luncheon attendees, 38 (59%) met the case definition. Persons who ate raspberries (relative risk [RR] = 5.4; 95% confidence interval [CI], 2.2-13.2) or potato salad (RR = 1.8; 95% CI, 1.2-2.6) were at significantly increased risk for illness. The population attributable risk percentages were 73% for raspberries and 20% for potato salad. Cyclospora oocysts were found in stools from 11 (85%) of the 13 case patients submitting specimens for testing. Implicated raspberries originated in Guatemala. CONCLUSIONS: Our investigation is one of the first studies to implicate a specific food (raspberries) as a vehicle for transmission of Cyclospora. Because of the apparent increasing incidence of cyclosporiasis in the United States, family physicians should consider testing for Cyclospora in any patient with prolonged, unexplained diarrhea. PMID- 9752378 TI - Ethnicity and rates of overweight. PMID- 9752379 TI - Domestic violence research. PMID- 9752380 TI - The biology of aging. AB - Various biological aspects of the aging process are reviewed. Aging has multiple causes. First, there is the initial genetic variability in the organism. Second, there is the destructive effect of the environment. Third, there is an accumulation in the somatic cells of changes, mutations or local chemical damage. These changes incapacitate or kill individual cells, and eventually cause a decline in physiological capacity. PMID- 9752381 TI - De senectute--a pediatrician takes a long look. PMID- 9752382 TI - Application of newer concepts in diabetes. AB - The changing definition of diabetes, brought up to date, includes the concepts of premellitus as well as mellitus, and of an enzymatically impaired or otherwise ineffective insulin as the possible cause of the full syndrome. Another current concept holds that an increased amount of this lipogenically active but glycolytically impaired insulin is the mechanism producing obesity in diabetes as a premellitic sign. Despite changed concepts the importance and effectiveness of prophylactic weight control in the premellitic stage, and as the fundamental treatment in all stages of obesity-diabetes remains the same. The order of preference in the addition of specific drugs to the dietary treatment of obesity diabetes is anorexigenic agents, phenformin, single sulfonylureas, and combined chlorpropamide-phenformin. Insulin is the therapy of last resort in this usually mild form of diabetes. There is great potential usefulness for oral hypoglycemic combination therapy (with steroids temporarily if necessary) as a replacement for insulin in the treatment of immunologically-produced insulin resistance. PMID- 9752383 TI - The kidney in hepatic disease and in gout: clinical and pathologic aspects. AB - The role of the kidney in hepatic disease and in gout has been surveyed. Kidney involvement is more common but less severe in hepatic disease, whereas it is less common but more severe in gout. The underlying pathology of the kidney lesions in both diseases (including studies by electron microscopy and immunofluorescence) is presented, with emphasis on the role this pathologic substratum plays in the renal insufficiency. The clinical aspects, including the diagnostic approach and therapeutic management of the "hepatorenal syndrome" and "gouty kidney" are discussed, as are the prognostic implications of the natural history of these "complications" and measures aimed at their prevention. PMID- 9752384 TI - Effect of antihypertensive (pargyline hydrochloride-methyclothiazide) therapy in three types of hypertension: preliminary report after one year of observation. AB - Fifty patients with elevated blood pressure were classified according to 3 sub groups as follows: 11 with borderline hypertension, 8 with systolic hypertension, and 31 with diastolic hypertension. So far, they have been observed for one year while being treated with an antihypertensive preparation containing pargyline hydrochloride and methyclothiazide. Response to treatment depended in large measure upon the type of hypertension; the borderline type was virtually unchanged; in the systolic type there was some diminution in the systolic, but less in the diastolic pressure; and in the diastolic type there was a reduction in both systolic and diastolic pressures. Side effects (faintness, nervousness, mouth dryness, insomnia, genitourinary disturbances and elevated blood uric acid level), when they occurred, were usually relieved by appropriate alteration of the antihypertensive drug dosage, by a change in the time of administration, or by adding medication directed at treatment of the side effect. Evaluation of cardiac output before and after therapy showed no change in this parameter. The results suggest: (a) that the antihypertensive effect probably was achieved by diminishing the peripheral resistance rather than by reducing the cardiac output, and (b) that there was no deterioration of myocardial efficiency, as measured by cardiac output, during the one-year period of antihypertensive therapy. More knowledge of the natural history of hypertension in each of the 3 sub-groups is required for better assessment of the influence of antihypertensive therapy on the outcome of the disease. Judgment as to the desirability of initiating therapy can in some measure be based on the classification of patients into appropriate sub-groups. PMID- 9752385 TI - Modern concepts in the management of emphysema in the aged and infirm. AB - The haphazard method of treating patients seriously ill with emphysema is gradually being replaced by modern, well organized cardiopulmonary therapeutic and rehabilitation programs. This evolution is the result of a further expansion in our knowledge of cardiopulmonary physiology and biochemistry; an additional factor is the recent public awareness of the seriousness of the problem, brought about by public educational programs. Emphysema is chiefly a disease of old age. It develops as a result of a degenerative process in which the alveolar walls become thinner and the lungs less elastic. Senile emphysema per se may not be disabling, but it is often associated with a severe chronic pulmonary disorder and thus can become the most distressing disease of old age, with shortness of breath even on such slight exertion as dressing or talking. Disturbances in cardiopulmonary physiology due to obstruction in the free flow of air and to superimposed infections require the use of all available procedures designed to obtain maximal pulmonary ventilation. The magnitude, the difficulty and the many controversial aspects of this problem are evident. The eventual solution will come gradually with continued interest and research. PMID- 9752387 TI - Atherosclerosis affecting vision: therapy. AB - The visual consequences and the pathogenesis and therapy of atherosclerosis are discussed. Atherosclerosis apparently is the result of hepatic failure to produce a stable suspension of cholesterol esters in the plasma. In some instances this represents an inherent metabolic defect. Usually it represents improper diet, namely, the excessive intake of saturated fatty acids and the inadequate intake of polyunsaturated fatty acids. Excessive ingestion of carbohydrates, insufficient physical exercise and reduced thyroid function are contributing causes. The final common denominators leading to tissue injury and destruction are vascular insufficiency and hypoxia. Hypertension is a separate disease, often concurrent with atherosclerosis. In diabetes, also a separate disease, atherosclerosis is one of the sequelae. Therapy primarily consists of the reduction or elimination of meat and milk fats from the diet, and the inclusion or increase of marine fats and vegetable oils. Simultaneously, carbohydrate intake is restricted. Adequate thyroid function, a normal hemoglobin level, and sufficient physical exercise are important. A supplementary intake of vitamins B6 and E, and of lipotropic substances is recommended. PMID- 9752386 TI - Patterns of water, ash and organic-matter changes in senescence. AB - A study of the changes in moisture, ash and organic matter in the organs of rats and mice has provided good evidence that there is a continuance of processes that occur at a constant rate throughout life. This rate is more rapid in the very early period of life, but the changes are related constantly to each other, as well described by certain mathematical equations. The loss of moisture is an hyperbolic function described by the logarithm of the percentage of concentration varying directly with the logarithm of the age of the animal. Similarly the concentration of ash declines while the total weight, total water and total ash increases by a complex exponential function described by a logarithm of the age in weeks. The gain in organic matter is best described by a power equation in which the logarithm of the percentage of organic matter is directly proportional to the logarithm of the age in weeks. These findings are in keeping with our previous suggestions and findings that constants of senile decay can be mathematically derived. It is further concluded that senility does not have a sudden onset, but is an accumulation of constantly related phenomena that can be accurately described mathematically. PMID- 9752388 TI - A vitamin, anabolic, stimulant mixture. Is this form of medication advantageous for debilitated geriatric patients? AB - Of 32 underweight geriatric patients in a double-blind study, 24 were given a therapeutic mixture containing, per dose, 10 mg of methylphenidate, 2.5 mg of methandrostenolone, and a mixture of B vitamins; 8 patients were given placebo only. After eight weeks of this therapy, there was a significant weight gain in the treated patients. In some cases, this was associated with increased bromsulphalein retention. The number of patients who showed clinically evident increases in strength and vitality was no greater than in the control group. The results lend no support to the use of tonic mixtures instead of separate drugs. PMID- 9752389 TI - High-level health for the elderly. AB - Human personality needs include survival, safety and security, acceptance, self esteem, and a sense of achievement. The fulfillment of these needs for the geriatric population requires greater community awareness of the potential value of these older people. PMID- 9752390 TI - A plan for regional medical health centers for the geriatric patient. AB - A rough plan is proposed which may facilitate the care of geriatric patients. The plan is to introduce pilot programs of geriatric regional centers attached to Veteran's Administration hospitals. Thus, at minimal cost, the problems of geriatric medicine would be under the supervision of our excellent university medical centers. PMID- 9752391 TI - [The nerve endings in contact with the parafollicular cells of the thyroid]. AB - We studied nerve endings in the parafollicular cells of thyroid glands in necropsy examinations of 50 young males with no know thyroid disorder. The Jabonero and Maillet techniques. ultrastructural and morphology study were performed together with a statistical analysis of nerve endings. Optic microscope visualized preterminal and exceptionally terminal nerve endings located near parafollicular cells. Swellings of the axons without Schwann's cell sheath were identified close to the par follicular cells with electronic microscopy, these swellings had an ellipsoidal or spherical shape and laid close to the cellular membrane (30 nm). Three types of synaptic-like vesicles were found in these nerve endings. a) Abundant clear vesicles, generally spherical in shape (average diameter 50.40 +/- 5.91 nm); b) A few large granural vesicles with an electron dense core (average diameter 105.49 +/- 9.98 nm); and c) Several small granular vesicles with an electron-dens core (average diameter 55.09 +/- 6.56 nm). Emphasis should be given to the finding of nerve endings (cholinergic-like, adrenergic-like and peptidergic-like nerve endings) in the parafollicular cells. We discuss their possible function. PMID- 9752393 TI - [Echocardiographic parameters in hyperthyroidism with and without cardiothyreosis]. AB - There has been little research done on the differences between hyperthyroidie patients who exhibit cardiothyrosis and those who do not. The objective of this research was to elucidate the variations in echocardiographic parameters that exist between these two groups, in order to determine practical implications. A prospective study on 37 subjects was performed: 20 in group I (without cardiothyreosis) and 17 in group II (exhibiting cardiothyreosis). In both groups, women were predominant. Left ventricular diameters and volumes were statistically higher in group II (p < 0.0007). The left ventricular end systolic stress was also higher (140.10(3) +/- 37 vs 131.10(3) +/- 51 dynes/cm2. p < 0.05). There was no significance between the two groups in shortening fraction, ejection fraction and mean rate of circumferential fiber shortening. The E/A ratio of the mitral flow was higher in group II (1.98 +/- 1.3 vs 1.3 +/- 0.7, p < 0.05), but the isovolumetric relaxation time and the deceleration time of the E wave were similar in both groups. Left ventricular systolic dysfunction was observed in 5 patients of each group. Relaxation abnormalities were found in 10% of the subjects in group 1 and 33.3% in group II. Significant valvular regurgitation was observed only in group II (8 cases of mitral, 1 case of tricuspid and I case of aortic regurgitation). Given that cardiovascular perturbations may be different during the stages of the hyperthyroidism, different therapeutic approaches may thus be considered, facilated by appropriate echocardiographic examination. PMID- 9752392 TI - [Thyroid function and autoimmunity in 215 patients seropositive for the hepatitis C virus]. AB - The aim of this study was to estimate thyroid function and the prevalence of thyroid antibodies among HCV seropositive patients. We undertook a screening for thyroid dysfunction, and antithyroperoxidase (ATPO) and antithyroglobulin (ATG) antibodies, in 215 HCV seropositive patients referred for hepatologic consultation, 118 males and 97 females, mean age 44 +/- 14 years, range 16-80 years. No patient was treated with interferon and all were seronegative for HIV. Eighteen patients (8%) had antithyroid antibodies, 12 with ATPO antibodies (5.6%) and 10 with ATG antibodies (4.7%). Four patients had both ATPO and ATG antibodies (1.8%), one case of Graves' disease and 3 cases of autoimmune hypothyroidism found during this study. Five patients (2.3%) had hyperthyroidism, three cases of Graves' disease, one case of iodine load and a case of Grave's disease incidentally diagnosed during medical examination. Eleven patients (5.1%) had hypothyroidism, one case already known and treated without antithyroid antibodies, 4 cases of autoimmune etiology (3 diagnosed in consultation and one already known and compensated hypothyroidism), one case of amiodarone-induced hypothyroidism discovered during this study, 5 cases of hypothyroidism without antibodies (two cases of compensated hypothyroidism with normal TRH stimulating test, two cases with severe liver cirrhosis and one case with chronic hepatitis). Twelve patients had antithyroid antibodies with normal TSH levels. The prevalence of ATPO and ATG antibodies in our study is similar to the prevalence usually observed in general population and does not suggest a pathogenic role of HCV in autoimmune thyroid disorders. PMID- 9752394 TI - [Primary hyperaldosteronism and adenoma of the adrenal cortex. Suppression of aldosterone with dexamethasone]. AB - The aim of the study was to evaluate dexamethazone test in a patient with primary aldosteronism caused by an adrenocortical adenoma. We observed a 50% decrease of plasma aldosterone as in glucocorticoid suppressible aldosteronism (GSA) but absolute value of aldosterone remained higher than 40 pg/ml. Basal plasma and urinary values of 18 OXO and 18 OH cortisol were not significantly elevated as in GSA. Inversely, the evaluation of 11 beta-hydroxylase activity of mineralocorticoids was in favor of a benign adrenal tumor. PMID- 9752395 TI - [Methylene blue in surgery of primary hyperparathyroidism]. AB - Is surgery for primary hyperparathyroidism easier when methylene blue (MB) is given preoperatively? This retrospective study compares the durations of interventions for primary hyperparathyroidism carried out after i.v. MB administration to those when no MB was given. Over a period of 20 years (June 1976 to December 1996), 175 consecutive patients (56 men and 119 women, with ages ranging from 16 to 92, mean 59.6) were operated upon for primary hyperparathyrodism; 55 were operated before February 1986--the period when BM was introduced routinely, and 120 after. Thirty-two other patients were excluded from the study: 14 had had a previous cervicotomy and 18 another procedure in addition to the parathyroidectomy (usually on the thyroid gland), two conditions which prolonged the time devoted to parathyroid identification and excision. Preoperative calcemia averaged 2.97 mmol/L (2.34 to 4.59) and mean preoperative PTH was equal to 2.6 times the upper normal limit (0.5 to 24.1). Both groups were similar for as age, sex, preoperative calcium and PTH, and histologies. Methylene blue was administered intravenously (5 mg/kg diluted in 500 cc of 5% glucose) over a period of time of one hour starting two hours prior to surgery. All 175 procedures were performed by two surgeons and duration of surgery was recorded from the anesthesiologist's notes. There were 149 adenomas (85%), 24 hyperplasias (14%), a combination of both in two, and unspecified in two others. Except for a case of acute lower back pain synchronous to the injection of the dye (which was immediately stopped), MB was well tolerated. Mean duration for the 55 interventions performed without MB was 68 minutes (35 to 140, median 60), compared to 49 minutes for the 120 procedures carried out after MB had been given (20 to 155, median 45). Differences in operative, times were highly significant (p < 10(-6) and represented a gain of time of 27%. Surgery for primary hyperparathyroidism was significantly shorter when it was preceded by the administration of MB, a dye which facilitates the identification of pathologic parathyroid gland(s). PMID- 9752396 TI - [Failure of synthcen to diagnose corticotropin insufficiency]. AB - It is commonly believed that a normal cortisol response to exogenous ACTH (synacthen test) reliably indicates, with the exception of states of acute ACTH deprivation, the integrity of hypothalamic-pituitary-adrenocortical function. Published evidence has shown that the synacthen test is not sensitive enough to reveal partial ACTH deficiency leading to a subnormal ACTH response to stress. We report a patient who maintained a normal response to exogenous ACTH stimulation despite symptomatic chronic ACTH deficiency in unstressed conditions. A normal cortisol response in the synacthen test does not therefore exclude the possibility of clinically relevant ACTH deficiency. PMID- 9752397 TI - [Commentary: choice of diagnostic tests for adrenal cortex insufficiency]. PMID- 9752398 TI - [Primary adrenal lymphoma with latent adrenal insufficiency: a case report and literature review]. AB - We report a new case of primary adrenal lymphoma with latent adrenal insufficiency and long-term remission after hydrocortisone replacement therapy. We have analyzed 29 other cases described in the literature. This disease with poor prognosis can be revealed by an incidentally discovered, frequently bilateral, adrenal mass. Adrenal insufficiency may be latent and the diagnostic procedure should include both cortisol and ACTH determination with an additional ACTH stimulation test if appropriate. Early adrenal substitution can improve patient survival. PMID- 9752399 TI - [Growth hormone replacement therapy: recommendations of the French Pituitary Club]. PMID- 9752400 TI - [Quality assurance approval in European health care centers training endocrinology, diabetes and metabolism specialists ]. PMID- 9752401 TI - [Why a "New Technologies" rubric in the Annals of Surgery?]. PMID- 9752402 TI - [Medico-economic evaluation of treatment of inguinal hernia: Shouldice vs. laparoscopy]. AB - Between May 1994 and September 1995, 64 men were included in a randomized prospective study comparing conventional Shouldice repair (S group) and transperitoneal laparoscopic repair with polypropylene mesh (L group). Cost evaluation was divided into distinct parts: drugs, non usable surgical materials, medico-technical procedures food and employees costs. In group S, mean operating time was 56', total cost was 3,922 FF in the case of unilateral hernia and 4,808 FF and respectively 77' in the case of bilateral hernia. In group L, mean operative time was 89', total cost 8,949 FF (disposable trocars) and 7,136 FF (non-disposable trocars) in the case of unilateral hernia and 116', 9,570 FF and 7,763 FF in case of bilateral hernia. Postoperative stay was 4.2 days in group S and 4 days in group L. Return to work was 28.6 days in group L and 35.5 days in group S (ns). In conclusion laparoscopic hernia repair does not decrease post operative pain, hospital stay and return to work, but is twice as expensive. PMID- 9752403 TI - [Radiotherapy of cancer of the anal canal. 15 years experience at the Lyon civil hospices and brief review of the literature]. AB - AIM: Retrospective analysis of a series of 287 anal canal carcinomas seen during a 15-year period in the Department of Radiotherapy Oncology of Hospices Civils de Lyon. MATERIAL AND METHOD: Between 1980 and 1995, 287 patients were managed in the radiotherapy department of Hospices Civils de Lyon. In 25 cases, the patients were referred for recurrence. Post or preoperative irradiation was performed in 12 and 23 cases. Palliative treatment was given to 15 cases and simple follow-up in 7 cases. Radiotherapy was given to 205 patients. In 71 cases irradiation alone and in 134 concomitant radiochemotherapy was given. RESULTS: The 5-year overall survival of the group treated by radiotherapy (205 patients) was 71.5% and the 10 year-overall survival was 60.8%. The specific survival at 5 and 10 years was 81.9% and 74.7% respectively. At 5 years the overall survival was 78% for the group treated by concomitant radiochemotherapy and 60% for the group treated by irradiation alone. CONCLUSION: Radiotherapy is the standard treatment for anal canal carcinoma. Radiochemotherapy seems to improve results in advanced cases. The best irradiation of technique has yet to be defined. PMID- 9752405 TI - [Laparoscopy-assisted hysterectomy and laparoscopic preparation. Apropos of a series of 177 cases]. AB - Our objective was to determine the limits of laparoscopic-assisted vaginal hysterectomy (LAVH) and the value of a preoperative scoring system to determine the operative approach to hysterectomy. Between January 1991 and December 1996, 152 out of 177 patients had LAVH and 25 had laparoconversion. The mean operating time was 163 min. The overall postoperative complication rate was 8.4%. The hospital stay was 4.8 days for LAVH versus 6.2 days for laparoconversion (p < 0.01). For each patient, a preoperative scoring system was established according to uterine size, previous laparotomy, uterine mobility, pelvic adhesions and endometriosis stage. The laparoconversion rate increased according to the score, as it was 7.8% for a score < or = 7 and 80% for a score > 7. LAVH offers a technique to convert some abdominal hysterectomies into vaginal hysterectomies. The use of the preoperative scoring system may help to determine patients who may benefit from the laparoscopic route and those with a high risk of laparoconversion. PMID- 9752404 TI - [Incidence and prevalence of colostomies. The future of stoma societies. Results of a Cote-d'Or survey. Federation of Stoma Patients in France]. AB - Few data are available in France about the impact of Ostomy Associations in the population of patients with permanent colostomy. The aim of this study was to assess the percentage of patients with permanent colostomy involved in local Ostomy Associations in a well-defined population (Cote-d'Or department, 493,931 inhabitants) in 1992. A minimum number of incident colostomy cases was computed using data from the Digestive Tract Tumor Registry in case of cancer and using the surgical reports from the University Hospital in case of benign disease. The maximum number of incident cases was computed using results from a survey conducted by the Public Administration (Social Security) during the same period. An interval of prevalent cases was assessed the number of collecting pouches based on sold during 1992 in the same area. A colostomy was set up in 37 to 84 patients. The number of prevalent cases was in a range of 274 to 410 patients. The corresponding percentages of Ostomy Associations members were included in a 8.3% to 19% range for incident cases and 9.5% to 19% for prevalent cases. These percentages are less than 30%, which is the presumed value previously evaluated in a survey conducted among physicians. As the incidence of colostomy cases continues to decline regularly, more effective methods must be developed to maintain a constant number of members. PMID- 9752406 TI - [Esophagectomy for epidermoid cancer discovered during evaluation of ENT cancer]. AB - The treatment of synchronous esophageal and head and neck carcinomas is difficult. MATERIAL AND METHOD: Retrospective study of 33 patients treated with esophagectomy for an intrathoracic squamous cells carcinoma discovered during pan endoscopy for a synchronous head and neck cancer. RESULTS: In 7 cases (21%) it was advanced (pT3-4) esophageal cancers. The hospital mortality was 9%. Five year survival was 18% without stabilization of the survival curve, 60% of patients died of recurrence of tumor. CONCLUSION: Esophagectomy is suitable for usT1-2 tumors if surgery is also indicated for the head and neck tumor. Radiochemotherapy is indicated for advanced usT3-4 esophageal tumors or when the treatment of the head and neck tumor is not surgery. PMID- 9752409 TI - [Using the electric scalpel in laparoscopy. Principles and prevention of complications]. PMID- 9752408 TI - [Surgical management of malignant bone tumors in the child. Saint-Etienne Hospital experience, apropos of 17 cases]. AB - From 1980 to 1997, seventeen children were treated for malignant bone tumors in the Department of Paediatric Surgery of the Saint-Etienne Hospital, with an average follow-up of 6 years and 3 months (2M-17Y). The histologic diagnoses were osteosarcomas (12 cases) and Ewing's sarcomas (5 cases). All children were treated surgically. For the 12 lower limb tumors, the techniques used were prosthetic replacement in 7 cases (associated with bone allograft in 3 cases), amputation in 3 cases and isolated bone allograft in 2 cases. The two ilium tumors were treated by resection and reconstruction with autologous bone graft. A single resection was performed for the shoulder-blade Ewing's tumors and a prosthesis for the 2 proximal humerus tumors. Six children died during follow-up. The authors stress that carcinologic resection is a priority before functional results. They insist on the mechanical failures of implants and on the difficulties of reoperations. PMID- 9752407 TI - [Laparoscopic treatment of pancreatic pseudocyst. 3 cases]. AB - Three patients with pancreatic pseudocysts were treated laparoscopically by cysto enteric by pass. Two developed a large cyst in the course of acute biliary pancreatitis. The third patient had chronic pancreatitis of unknown etiology. The mean size of the collection was 12 cm (8-20). Treatment was performed with a delay of 80 days (30-300) after the onset of the disease. Two patients underwent laparoscopic cysto-gastrostomy using laparoscopic ultrasound. Stapling anastomosis appears easy and safe. The third patient underwent operating laparoscopically assisted cysto-jejunostomy with an-Y loop. Mean time was 120 minutes (90-200) and hospital stay was 7 days (5-8) without morbidity. The follow up was 18 months without radiological recurrence and with excellent clinical results. PMID- 9752410 TI - [Anatomy of the liver and hepatectomy techniques]. PMID- 9752411 TI - [Benign tumors of the liver: primum non nocere]. PMID- 9752412 TI - [Biliary cysts and hepatic polycystosis]. PMID- 9752413 TI - [Hydatid cyst of the liver]. PMID- 9752414 TI - [Local, non-surgical treatment of hepatocellular carcinoma]. PMID- 9752415 TI - [Surgical treatment of hepatocellular carcinoma]. PMID- 9752416 TI - [Liver imaging for metastases]. PMID- 9752417 TI - [Surgery of liver metastases of colorectal carcinoma]. PMID- 9752418 TI - [Surgery of liver metastases of non-colorectal origin]. PMID- 9752419 TI - [Malignant melanoma of the uterine cervix. Apropos of a case, with total colpo hysterectomy and vaginal reconstruction using a rectus abdominis flap]. PMID- 9752420 TI - [Angiomyolipoma of the liver]. PMID- 9752421 TI - [Ciliated cyst located in the common bile duct]. PMID- 9752423 TI - [Combined locoregional therapeutic strategy in locally advanced cancers of the pancreas]. PMID- 9752422 TI - [Biliary ileus. Apropos of 7 cases]. PMID- 9752424 TI - [Ovarian teratoma prolapsed into the rectum]. PMID- 9752425 TI - [Primary retroperitoneal hydatid cyst]. PMID- 9752426 TI - [Gilles Bousquet. (1936-1996)]. PMID- 9752427 TI - ["History of Medicine" rubrics in our scientific journal]. PMID- 9752428 TI - [Surgery of morbid obesity: intestinal bypass to adjustable gastric banding]. AB - The authors present their experience at the Centre for the surgical treatment of morbid obesity at Milano University where since 1974, 603 obese patients underwent surgery: 312 jejuno-ileal bypass (JIB), 70 bilio-intestinal bypass (BIB), 102 horizontal gastroplasties (HGP), 44 silastic ring vertical gastroplasties (SRVGP) and 75 adjustable silastic gastric banding (ASGB). Average follow-up for these procedures is 16, 6, 11, 4 years and 24 months respectively. Weight loss is satisfactory in all cases even though the percentages vary in the different procedures. The most serious complications (severe hepatic failure, oxalic interstitial nephritis, persisting malabsorption) occurred in patients submitted to JIB. The best clinical outcome with the lowest complications rate was obtained with BIB compared to other intestinal bypasses. The most frequent complication observed in patients submitted to gastroplasties was incoercible vomiting while the most severe complications were diffuse peritonitis, secondary to gastric perforation, and peripheric neuropathy. Our experience confirms that surgical treatment of morbid obesity refractory to medical therapy is today a safe and effective treatment. BIB has still a role in super-obese young patients (BMI over 50) refusing dietary restriction lifetime. The gastric procedures, especially laparoscopic ASGB, seem to be the best option. The excellent outcome of bariatric surgery can be obtained only in specialized centers where various specialists work together. PMID- 9752429 TI - [Inguinal hernia. 4-year follow-up of 2 comparative prospective randomized studies of Shouldice and Stoppa operations with pre-peritoneal totally laparoscopic approach (461 patients)]. AB - The aim of this study was to evaluate the late (4 years) recurrence rate after laparoscopic totally preperitoneal (TPP) approach, to Shouldice and Stoppa procedures. All patients were reviewed at one, six months, one year and yearly there after. The mean follow-up was 4 years: the follow-up was 100 per cent at one month, 98 per cent at 6 months, 95 per cent at one year, 91 per cent at 2 years, 84 per cent at 3 years, 79 per cent at 4 years and 61 per cent at 5 years. At one year, the recurrence rate was 2.2 per cent in the laparoscopic group, 1.2 per cent for Shouldice and 0 per cent for Stoppa procedure (ns). At 3 years, the recurrence rate was comparable and 3.6 per cent for laparoscopy, 5.1 per cent for Shouldice and 5.2 per cent for Stoppa respectively (ns). At 4 years, the recurrence rate was lower (but ns) for the laparoscopic group 7.4 per cent versus 12.5 and 10.5 per cent. Two predictive factors for recurrence in laparoscopic treatment were the size of the mesh and the surgeon's experience. PMID- 9752430 TI - [Value of celioscopy in treatment of isolated torsion of the Fallopian tube. Review of the literature. Apropos of 3 cases]. AB - Isolated fallopian tube torsion (ITT) is infrequent and associated with morphologic and dynamic disturbances. Mr L, 31 years old, suffered from right lower quadrant pain which became worse during the following 48 hours. Laparoscopy revealed a right necrotic ITT which was resected by laparotomy. Mr L, 49 years old, suffered from by left lower quadrant pain with progressive onset. Laparoscopy revealed a left necrotic ITT which was resected. Mr P, 76 years old, suffered from left lower quadrant pain for 14 days. Ultrasonography revealed an adnexal mass. Laparotomy revealed a left necrotic ITT which was resected. On literature review, ITT (81 cases) was revealed by lower quadrant pain, acute onset, which quickly became worse. Pelvis examination revealed a lateral cul-de sac pain. Ultrasonography identified tubal cystic mass with high-impedance arterial waveform on colour Doppler sonography. Diagnosis was easily established by laparoscopy. In case of clinical symptoms suggestive of ITT, pelvic and endovaginal ultrasonography and laparoscopy are indicated. Tubal preservation must be the rule. PMID- 9752431 TI - [Gynecomastia. Management of diagnosis and therapy. Apropos of 52 cases]. AB - Gynecomastia is the commonest breast lesion in males. Fifty-two patients (mean age 24 years) operated in our department were reviewed with a mean follow-up of two years and a half. Gynecomastia occurred most frequently during puberty (63%), was bilateral (75%) and idiopathic (65%). The size of the enlargement was evaluated according to Simon's-classification based on breast-volume and skin redundancy. 18 stage 1, 22 stage 2A, 9 stage 2B, 3 stage 3. Clinical examination and mammography determined the consistency of gynecomastia: adipose or firm. 4 different surgical managements were used: 32 subcutaneous mastectomies, 12 liposuctions, 6 liposuctions assocaited with subcutaneous mastectomy, 1 total mastectomy. One patient had liposuction on one side and subcutaneous mastectomy on the other one. All techniques gave good morphologic results. Nonetheless, the authors recommend the combination "liposuction and subcutaneous mastectomy", as this technique presents many advantages: small intraoperative blood loss, good skin redraping, short hospital stay, complete histologic examination of the material removed. PMID- 9752432 TI - [Surgical incisions in France in the 19th century]. PMID- 9752434 TI - [Blunt injuries of the liver. Reliable approaches to treatment]. PMID- 9752433 TI - [Current diagnostic and therapeutic approach to common bile duct calculi: a rapidly evolving field]. PMID- 9752435 TI - [Intrahepatic gallstones]. PMID- 9752436 TI - [Bile duct cysts]. PMID- 9752437 TI - [Biliary cholangiocarcinoma]. PMID- 9752438 TI - [Biliary emergencies in the era of celioscopy]. PMID- 9752439 TI - [Combined radiochemotherapy in treatment of esophageal cancers]. AB - Concomitant radiochemotherapy has totally changed the treatment of oesophagus cancer. After numerous phase II studies that indicated their feasibility and efficacy, randomised phase III studies recently demonstrated their ability to improve survival. In operable cancer, preoperative radiochemotherapy increases the overall survival in adenocarcinoma, and the disease-free survival in squamous cell cancers, in comparison to surgery alone. In locally advanced disease, radiochemotherapy is considered as standard treatment. At this moment the effective drugs are 5-fluorouracil, mitomycin C and cisplatinum. The aims of current studies are to increase further the therapeutic ratio, the ways being refinements of radiotherapy and new chemotherapy delivery modalities. Finally the efficacy of radio-chemotherapy is questionning the role of surgery in this disease. PMID- 9752440 TI - [Pleuro-peritoneal Denver shunt in treatment of chronic pleurisy]. AB - The first utilisation in our hospital of a pleuraperitoneal shunt for the treatment of chronic pleurisy enabled us to study, in the light of published data, the place for such a procedure in the management of resistant pleural effusion. The aetiology of the pleurisy in a 70-year old patient who underwent this mini-invasive surgery was unknown when the shunt was inserted, but his symtoms clearly improved afterwards. At the present time with a follow-up of 13 months, there are no local complications and the system is in good working order. In 1982, the material used for the first time in such a case was an adapted version of Denver's peritoneal venous shunt. This is composed of a pleural catheter linked by a pump that the patient controls himself, to a peritoneal catheter. This pump can be inserted under local anaesthetic. The principal indications in the literature, in which the series do not exceed 70 cases are: malignant pleurisy where it is preferable to introducing tale in the case of tissue retraction fastening to the underlying lung and also in chylous pleurisy, especially those secondary to congenital east disease in children. A complication rate of 25% is noted depending on the type of infection or obstruction leading to replacement of the shunt. No case of erosion has been noted. The long term patency, measured by radio-isotope injections (Tc99m), has not been studied but there is a significant reduction in the length of hospital stay which gives a clear economic advantage to such procedures. PMID- 9752441 TI - [Metastatic vipoma. Could an aggressive therapeutic approach improve survival?]. PMID- 9752442 TI - [Retro-esophageal subclavian artery. Apropos of 2 cases]. PMID- 9752444 TI - [Apropos of a penetrating wound of the common bile duct caused by a knife]. PMID- 9752443 TI - Intercostal transdiaphragmatic hernia with abdominal contents. PMID- 9752445 TI - [Volvulus of the cecum: treatment by resection]. PMID- 9752446 TI - [Dorso-lumbalgia and colorectal pathology]. PMID- 9752447 TI - [Gastric banding in treatment of morbid obesity. Experience with 51 cases]. PMID- 9752448 TI - [Intraductal papillary mucinous tumors of the pancreas: are there any preoperative clinical and laboratory factors predictive of degeneration. Results of a French-Belgium collective series]. AB - Intraductal papillary and mucinous tumors are rare. We retrospectively analysed clinical, surgical and histological features and outcome of 41 operated patients (29 males, 12 females, mean age = 63 years). The commonest presenting manifestation was acute pancreatitis (41%). Tumor was located in only one pancreatic segment in 45% cases. Forty one per cent of patients had invasive carcinoma, 20% had tumor with severe dysplasia and 39% with minimal or moderate dysplasia. Only elevated age was significantly associated with invasive carcinoma. Eleven out of 17 patients with invasive carcinoma (65%) had a recurrence after surgery and 6 (35%) died. Among 24 patients with noninvasive tumor, 2 (8%) recurred without tumor-related death in the follow-up (48 months). This study underlines the need for early surgical resection in patients with intraductal papillary and mucinous tumor because of the high frequency of invasive carcinoma and the poor outcome of patients with invasive carcinoma. PMID- 9752449 TI - [Results of laparoscopic treatment of abdominal emergencies]. AB - The purpose of this retrospective study was to evaluate the results of the laparoscopic surgical treatment of abdominal emergencies. From May 1991 to September 1995, 200 patients operated by laparoscopy for an acute abdomen were included in this study. The decision to treat the patient by laparoscopy was taken by the surgeon on duty. There were 101 males and 99 females with a mean age of 41 +/- 20 years (range 11-90 years). The main indications for operation were: acute appendicitis (109 patients), acute cholecystitis (52 patients), small bowel obstruction (14 patients) and perforated duodenal ulcer (14 patients). There was no hospital mortality. One per cent of patients experienced an operative complication which was treated by laparotomy. Conversion to laparotomy was needed in 13% of cases. The morbidity rate was 9% and reoperation by laparotomy for acute generalized peritonitis secondary to small bowel perforation was necessary in two cases. Mean postoperative hospital stay ranged from 4 to 7 days. The authors conclude that surgical laparoscopic treatment of the common abdominal emergencies is safe. The conversion rate is low as is the complication rate. These conclusions should be confirmed by a prospective study. PMID- 9752451 TI - [Surgical management of adnexal tumors]. AB - Concerning laparoscopy and tumor dissemination, we know from several multivariate analyses that at laparotomy, if the tumor is entirely and immediately removed, the puncture of a stage I ovarian cancer has no influence on the prognosis. In contrast the inadequate surgical management of an undiagnosed ovarian cancer may worsen the prognosis. The diagnosis is the key step. To be able to immediately and completely treat an ovarian cancer when managing an ovarian tumor surgically, Laparoscopic diagnosis is safe and reliable when used cautiously. The surgical diagnosis may and should probably be performed by laparoscopy whatever the ultrasonographic appearance of the tumor. Masses diagnosed as suspicious at surgery should be treated by immediate laparotomy, since the results of laparoscopic treatment of an ovarian cancer are not known. In young patients, conservative surgery is the main advantage of laparoscopy, and should be achieved in most benign masses. The recent progresses of in vitro fertilization should be taken into account when managing an ovarian tumor in a patient who wishes to become pregnant. Frozen sections are useful, when treating highly suspicious masses, allowing an immediate staging and avoiding the disadvantages of a second surgical procedure. Whenever a malignant tumor has been missed at laparoscopy, restaging is required and should be considered to be an oncologic emergency. PMID- 9752450 TI - [Effect of preoperative administration of Lugol's solution on thyroid blood flow in hyperthyroidism]. AB - A study of 50 patients with hyperthyroidism was conducted to evaluate the effect of preoperative administration of Lugol's iodine solution on thyroid blood flow. Highly significant reductions in diameter, time-averaged velocity, and volume flow of the superior thyroid artery were demonstrated after administration of Lugol's solution. The Duplex ultrasound scanning used in this study is a noninvasive, inexpensive, accurate, and reproducible technique suitable for analysis of thyroid blood flow in hyperthyroidism. On the basis of current ultrasonographic results and low postoperative morbidity in patients, Lugol's solution is well tolerated and may be recommended for use before thyroidectomy, especially for diffuse toxic goiters and Graves disease. PMID- 9752453 TI - [Centenary session of the Surgical Society of Lyons. Inaugural address]. PMID- 9752452 TI - [Interest in CD-ROM for surgical teaching]. PMID- 9752454 TI - [Gynecological surgery in hospitals in Lyons, France, from 1897 to the present]. PMID- 9752456 TI - [The evolution of gastrointestinal surgery over the last hundred years]. PMID- 9752455 TI - [One hundred years of bone surgery in the Lyons Teaching Hospitals (1897-1997)]. AB - Throughout the XIXth century and until 1945, bone surgery focused primarily on correcting deformities in children. The treatment of injury-related bone lesions in adults (compound fractures and dislocations) remained within the province of general surgeons until circa 1970. Lyons played a unique role in the XVIIIth and XIXth centuries, for three reasons. 1) The term "orthopedics" (which means "straight children", or "children to be made straight") was coined in 1743 by an 80-year-old inhabitant of the Saint Nizier parish in Lyons, Nicolas Andry or Andre, a former dean of the Paris School of Medicine. The term was adopted throughout the world. 2) The first French orthopedic surgeon was Gabriel Pravaz, who was from the Pont-de-Beauvoisin neighborhood of Lyons. He treated "children to be made upright" in his orthopedic and pneumatic institute located quai des Etroits, and was the first to successfully reduce congenitally dislocated hips (before 1850). 3) In 1987, the most famous bone surgeon was Ollier, who brilliantly applied the method developed by Claude Bernard to the experimental study of the role of the periosteum. He observed that new bone was laid down after "subperiosteal resection" of infected bone or joint tissue. His technique allowed to reduce dramatically the number of limb amputations for infection. In 1897, in addition to Ollier (who died in 1900), many other outstanding surgeons from Lyons demonstrated an interest in bone surgery. Around 1897. Jaboulay (known as a vascular and transplantation surgeon) successfully performed amputations through the middle of the pelvis. Also during this period, Gangolphe used osteotomies to correct limb deformities, and also performed bone grafts. Between 1897 and 1910, the radiologist E. Destot, who worked at the Hotel-Dieu hospital in Lyons, published two books on the classification of fractures, "The wrist" and "The foot", both of which were promptly translated in English. The Lyons School at the Hotel-Dieu that demonstrated its excellence in the XIXth century gave birth in the XXth century to a school of infantile bone surgery, headed by G. Nove-Josserand from 1894 to 1937 then by L Tavernier from 1937 to 1947. Tavernier was a sports enthusiast, and was proficient not only in pediatric surgery but also in the areas of meniscal lesions and bone tumors. He was succeeded by Guilleminet (1947-1962), then by Joseph Marion. Two teaching hospital departments of adult orthopedic surgery were created, one headed by Albert Trillat, who, together with H. Dejour and A. Mounier-Khun, was known throughout Europe as an outstanding sports injury surgeon, and the other by Jean Creyssel, who worked with G. de Mourgues and played a significant role in France in bringing trauma related injuries (traditionally treated by general surgeons) into the field of orthopedic surgery. The first successful total hip arthroplasty procedures were done around 1966. Orthopedics and traumatology became a separate specialty in 1969, and in the wake of this change many departments focusing only or preferentially on one part of the body (e.g., the hand and upper limb, knee, or hip) were created. Over the last century, the Lyons Society for Surgery has played a key role in publishing discoveries in bone surgery and in disseminating knowledge in this field. Although societies for surgery of the hip, knee, spine, hand, foot, and so on now exist, meetings of the Lyons Society for Bone Surgery remain useful since new ideas and techniques sometimes stem from experience acquired in other fields. It is worthy of note that in other European countries traumatology is a specialty in itself, which includes visceral and bone traumatology. It can be anticipated that harmonizing the traumatology specialty in Germay and the orthopedic surgery and traumatology specialty in France may raise a number of problems. PMID- 9752457 TI - [Urology and transplant surgery]. PMID- 9752458 TI - [One hundred years of heart surgery]. PMID- 9752459 TI - [One hundred years of neurosurgery in Lyons: the milestones]. PMID- 9752460 TI - [Laparoscopic discovery of disseminated peritoneal leiomyomatosis closely resembling metastatic peritoneal carcinomatosis]. PMID- 9752461 TI - [Malignant oncocytoma of the thyroid presenting with hepatic metastasis]. PMID- 9752462 TI - [Prevention, detection and management of colon cancers]. PMID- 9752463 TI - [Shouldice steel wire repair and under local anesthesia: prospective evaluation of postoperative comfort]. AB - This study evaluates postoperative comfort of patients who have undergone inguinal hernia operations by Shouldice technique. The low analgesic consumption, the out patient management of 77% of patients, the resumption of normal activity after one week and a satisfaction index of nearly 100%, are the main results of this study. For the authors, this comfort is another argument in favour of the use of this technique. PMID- 9752464 TI - [Appendicular pseudo-tumors: unusual diagnosis]. AB - The discovery of a tumour mass of the appendix, in an acute or chronic context, raises the problem of its benign or malignant, inflammatory or infectious nature. We report five cases of patients operated by the same surgical team between June 1991 and September 1996, who presented macroscopically and histologically with unusual appendicular pseudotumours: appendicular diverticulosis (n = 1), Crohn's disease localized to the appendix (n = 2), yersiniosis (n = 1), actinomycosis (n = 1). The preoperative diagnosis was acute appendicitis (n = 2) or tumour (n = 3). The postoperative course was uneventful in every case, and specific medical treatment was prescribed in two cases (yersiniosis and actinomycosis). These differential diagnoses must be considered in all appendicular diseases, but they are extremely difficult to confirm preoperatively. PMID- 9752465 TI - [Approach to upper limb edema secondary to subclavian vein occlusion situated distal to an arteriovenous fistula]. AB - AIM OF THE STUDY: To analyse the course of upper limb edema in patients with an arteriovenous fistula used for dialysis and to analyse the available therapeutic options. STUDY DESIGN: Retrospective study of patients with this type of edema, who were treated in our institution from 1992 to 1996. PATIENTS AND METHODS: Seven consecutive patients with an arterioveinous fistula treated for edema of the upper extremity, were reviewed. The fistula was created at the elbow in 6 patients and at the forearm in 1. The edema appeared immediately after operation in 4 patients and after a delay in 3 patients. Stenosis (3 patients) or occlusion (2 patients) of the subclavian vein was documented in 5 patients who were investigated by angiography. RESULTS: The edema regressed spontaneously in 4 patients because collaterals developed in 3 patients, and the fistula thrombosed in 1 patient. Surgical intervention allowed regression of the edema in the other 3 patients: excessive output of the fistula was reduced in 2 patients and an axillojugular bypass was performed in 1 patient. The fistula remained effective in 6 patients. Another fistula was performed on the contralateral arm in 1 patient. CONCLUSION: Non-operative management is recommended in patients who develop edema immediately after creation of the fistula, because spontaneous regression is likely. Measures aimed at reducing the output of the fistula or enhancing the venous capacities of the arm are required when edema appears at a later stage. The fistula can be saved in the majority of cases. PMID- 9752466 TI - [The origins of the National Academy of Surgery]. AB - The French Academy of Surgery was not created easily largely due to the frequent quarrels between physician and surgeons. Founded in 1731, it experiented many trials and tribulations for three centuries. Disbanded in 1793 by the Convention, it was transformed into Societe Nationale in 1843 and only regained the name of Academie in 1935. PMID- 9752467 TI - [Exclusive use of calcium channel blockers and cardioselective beta-blockers in the pre- and per-operative management of pheochromocytomas. 70 cases]. AB - The pre and intraoperative use of calcium channel blockers (CCB) has been suggested for the management of either eutopic or ectopic pheochromocytomas. We report our experience of 70 pheochromocytomas, operated between 1988 and 1996 and managed with CCB, especially nicardipine. 59 were hypertensive (84.2%). Preparation consisted of nicardipine in 61 patients or another CCB in 9 cases with duration ranging from 24 hours to several weeks depending on plasma volume and blood pressure control. Intraoperatively, nicardipine infusion was started after intubation, adjusted according to systolic blood pressure (SBP) and stopped before ligation of the tumor venous drainage. Increases in SBP greater than 200 mmHg were observed in 10 patients and were effectively controlled by nicardipine in all cases. In 16 patients, the S > BP remained less than 150 mmHg throughout anesthesia. Heart rate greater than 100 b p m occureed in 51 patients and was easily controlled with esmolol whenever used (n = 27). Arythmias were unfrequent (n = 4) and required treatment in only one case. This study confirms the ability to adequately manage pheochromocytomas with the use of nicardipine as sole vasodilating agent. PMID- 9752469 TI - [Bilateral video-endoscopic adrenalectomy in Cushing's disease. Experience in 24 patients]. AB - The purpose of this study was to compare the results of bilateral laparoscopic adrenalectomy (BLA) to bilateral open adrenalectomy (BOA) in the treatment of Cushing's disease. Twenty-four patients (23 Cushing's disease, 1 congenital adrenal hyperplasia) were divided into 3 groups. Group 1 patients (n = 15) underwent BCA using the lateral transabdominal approach, Group while 2 patients (n = 9) underwent laparoscopic adrenalectomy on one side and conventional open adrenalectomy on the contralateral side. Groups 1 and 2 were compared retrospectively to 15 patients (Group 3) who underwent BOA as part of larger series of 61 patients. There was no difference in the degree of hypercortisolism in the 3 groups. At the beginning of the experience, the duration of surgery was longer in Groups 1 and 2 compared to the open surgery group, but this difference subsequently decreased during the study. There was no difference in intraoperative blood loss or transfusion rate. Group 1 patients experienced fewer wound and intraabdominal complications and less postoperative pain, shorter hospitalization, and quicker recovery than groups 2 and 3 patients. Technically obesity and tissue fragility are easily overcome by the laparoscopic approach. BCA also achieves success rate of hypercortisolism correction. In conclusion, BLA is the surgical procedure of choice for the treatment of Cushing's disease when surgical therapy is indicated. PMID- 9752468 TI - [Comparative study between laparoscopic and conventional adrenalectomy for pheochromocytomas]. AB - This prospective study compares the outcome of 2 groups of patients with pheochromocytoma undergoing adrenalectomy via a transabdominal approach or a laparoscopie approach. Mean operating time was exactly the same in both groups (122 and 125 minutes). Postoperative stay in the intensive care unit and in hospital was significantly shorter in the laparoscopic group as previously demonstrated fo other adrenal tumors. More important, no differences in haemodynamic changes were observed during surgery. In both groups, the increase of cathecholamine levels was identical, with a peak occuring during manipulation of tumors. Pneumoperitoneum was not responsible for any cardiovascular instability. Laparosopic adrenalectomy should therefore be preferred to open procedures in pheochromocytomas unless tumors are too large or potentially malignant. PMID- 9752470 TI - [Malignant cortico-adrenal tumors with vena cava extension. Is surgical resection justified?]. AB - Between January 1989 and December 1996, 6 patients (4 females and 2 males), ranging in age from 23 to 82, underwent surgery for malignant adrenocortical tumors with inferior vena cava extension. There were 5 adrenocortical carcinomas and 1 metastasis from a differentiated thyroid carcinoma. The tumor size ranges from 9 to 17 cm. 5 tumors involved the right adrenal and 1 involved the left adrenal. Four patients had a vena caval thrombus, extending into the right atrium in one case. Two patients had direct tumoral invasion of the IVC wall without thrombius. All patients underwent en bloc excision of the adrenal gland and regional nodes, with nephrectomy in 4 cases and right hepatectomy in one case. Thrombectomy was performed in 4 cases with cardio-pulmonary by-pass in one case. Venous invasion without thrombus required total resection of the infra-hepatic IVC ine one case and vascular reconstruction with prosthetic patch in another case. There were no peri-operative deaths. Two patients died from metastatic disease after 13 and 40 months respectively. Four patients were alive with a follow-up ranging from 3 to 31 months, 3 patient were free of disease. Surgical resection of adrenocortical carcinomas with IVC extension can be attempted. There is no increase in morbidity and mortality in comparison with surgical treatment of other adrenal carcinomas. In these particular cases, the risk of metastases and local recurrence is high, but long-term survival can be obtained after radical macroscopic resection. PMID- 9752471 TI - [Is laparoscopic resection of a malignant corticoadrenaloma feasible? Case report of early, diffuse and massive peritoneal recurrence after attempted laparoscopic resection]. AB - As surgeons wholeheartedly and intensely supporting (and deeply involved in) laparoscopic adrenal surgery, we think that laparoscopic resection of benign adrenal tumors is about to make conventional laparotomy obsolete. However, considering the many reported cases of tumor seeding after laparoscopy, we consider that proven or suspected adrenal cancer remains an absolute contra indication for laparoscopy. It is precisely for this reason that we feel duty bound to report the tragic and life-threatening case of a 25-year-old woman operated for malignant adrenal cancer and suffering from diffuse and massive peritoneal carcinomatosis diagnosed at the sixth postoperative month. A complete macroscopic debulking was carried out with removal of all visible tumor nodules followed by immediate intraperitoneal cisplatinum chemotherapy and six postoperative courses of systemic cisplatinum, etoposide and mitotane chemotherapy. This combined and aggressive therapy induced remission that perhaps could have been initially obtained with a more critical and proper surgical operation. We hope this paper will contribute in the future to avoiding similar catastrophes that could rapidly discredit such a promising new form of surgery. PMID- 9752472 TI - [Pancreatic echo-endoscopy and preoperative localization of insulinomas]. AB - Preoperative localization of insulinomas often fails because of the small size of these tumors. The aim of this study was to analyse the value of endoscopic ultrasonography of the pancreas in comparison with them to those of conventional localization procedures. PATIENTS AND METHODS: From 1983 to 1997, 32 patients, operated with a preoperative diagnosis of insulinoma, underwent one or more localization procedures: ultrasonography (US) (n = 31), computed tomography (CT) (n = 31), magnetic resonance imaging (MRI) (n = 10), angiography (ANG) (n = 6), transhepatic portal venous samplings (THPVS) (n = 3), and/or endoscopic ultrasonography (EUS) (n = 25). More recently, 4 patients had scintigraphy with labelled octreotide. During surgery, intraoperative palpation and ultrasonography of the pancreas, performed in all but one cases (laparoscopy), allowed the localization of 29 solitary tumors and 2 multiple tumors (one of which in a case of a MEN II). A malignant tumor was found in 6 patients. RESULTS: The sensitivity of the localization procedures was as follows: US = 19%, CT = 39%, IRM = 30%, ANG = 33%, THPVS = 0%, EUS = 96%. Labelled octreotide scintigraphy was positive in 3/4 cases. Surgical procedures included: 15 enucleations or partial resections, 14 left pancreatectomies (5 of which with splenectomy), 3 duodenopancreatectomies. In one case the tumor was resected laparoscopically (distal pancreatectomy). CONCLUSION: EUS was the best preoperative localization procedure in this study. It may avoid the need for other imaging procedures. Combined with intraoperative ultrasonography, EUS could allow laparoscopic resections in selected cases. PMID- 9752473 TI - [Limits of parathyroid scintigraphy before surgery for hyperparathyroidism of renal origin]. AB - AIM OF THE STUDY: To evaluate the results of parathyroid scinti scans (sestamibi or tetrofosmin) for detection of hyperplastic parathyroid glands responsible for renal hyperparathyroidism. METHODS: Injection of 15 mCi sestamibi or tetrofosmin and gammacamera acquisition of images focused on neck and mediastinum, 20 minutes and 2 hours thereafter. Injection of 150 mCi Iodine 123, acquisition of images 2 hours afterwards and visual subtraction. PATIENTS: 51 patients with renal insufficency or renal transplant were referred for surgical treatment of hyperparathyroidism. 52 scintiscans (sestamibi n = 19, tetrofosmin n = 33) were performed before operation (subtotal parathyroidectomy, bilateral thymectomy and parathyroid tissue cryopreservation). RESULTS: 180 hyperplastic parathyroid glands were resected, 71 of which had been detected by scintiscan. The factors modifying the results were the weight of the resected lesion and reoperation. All hyperplastic glands were detected in only 1 out of 41 scintiscans performed before first hand operations, whereas all missed glands were imaged in 8 out of the 10 explorations performed before reoperation for persistent renal hyperparathyroidism. The radionuclide, the type of hyperparathyroidism, the parathyroid location, patient's age and gender did not influence the results. No false-positive result was observed. CONCLUSION: Parathyroid scintiscan should not be routinely performed before the first neck exploration for renal hyperparathyroidism. It is mandatory in those cases needing reoperation for recurrent disease. PMID- 9752474 TI - [Mechanical value of prostheses and fixation with staples in repair of abdominal eventrations]. AB - From an experimental study on fresh corpses, using a system which allows reproductible measures, we evaluated the physical strain to which prosthetic meshes are subjected during the initial phase of repair of abdominal eventrations treated by mesh. We also studied the various systems of staple fixation. The adhesivity or resistance-to-tear of the mesh is minimally dependent on the texture of the plaque; additionall, we also demonstrated the role of the size of the mesh. Stapling may compensate for the lack in size and increase the resistance-to-slip with larger values for radial stapling as compared to tangential stapling. Other fields of applications are possible. The use of glue or absorbable staples is considered. PMID- 9752476 TI - [Torsion of a supernumerary spleen. Case report]. PMID- 9752475 TI - [Winslow's foramen hernia. Preoperative computed tomographic diagnosis and laparoscopic treatment]. PMID- 9752477 TI - [Current management of Barrett's esophagus]. PMID- 9752478 TI - [Telemedicine applied to surgery]. AB - The transmission of images, audio, video and computed data allows new possibilities in initial or continuing education/training of surgeons and new interactivities between medical or surgical specialties. In the field of surgery, telemedicine is used for teaching, diagnostic or therapeutic assistance, and even consultation of remote patients. This article reviews telemedicine technologies, national legislation current, surgical telemedicine practices and the future applications (telementoring, surgical simulator, telesurgery...). PMID- 9752479 TI - [Impact of tumor doubling time on the therapeutic strategy: application to so called synchronous metastases of colorectal cancers]. AB - An exhaustive review of tumor doubling time (TDT) is presented, showing that growth of a solid malignancy remains subclinical for three-quarters of its duration and that tumor growth is close (but not strictly identical) to an exponential type. The TDT can be transiently accelerated after an aggression, particularly after surgery (which induces immunodepression and stimulation of growth factors during healing). In contrast, the TDT can be slowed by effective chemotherapy. Based on these elements, the classical attitude of waiting 3 to 6 months before resecting synchronous liver metastases from a colorectal primary is illogical, as it has only a 5 to 10% chance of detecting new metastases during this interval. Therapeutic decisions must therefore be exclusively based on surgical technical considerations, rather than erroneous oncologic considerations. In terms of the tumor growth, it is better to operate once rather than twice. In terms of the patient's quality of life and socio-economic aspects, one operation is also preferable to two. Resection of the primary tumor and synchronous metastases should therefore be performed during the same operative procedure whenever possible. PMID- 9752480 TI - [Treatment of esophageal adenocarcinoma]. AB - Surgical treatment of adenocarcinoma of the esophagus is of topical interest because an increasing rate of incidence. The main purpose of this study was to determine the surgical management of adenocarcinoma in Barrett's esophagus, non Barrett's adenocarcinoma, early adenocarcinoma, high-grade dysplasia in Barrett's esophagus and to precise whether multi modality therapy after a survival advantage. PMID- 9752482 TI - [Pseudo-acute surgical abdomen and acute leukemia]. AB - The so-called pseudo-acute abdomen has been reported in acute leukemia, both at diagnosis or relapse. The clinical presentation may be misleading and life threatening, due to the possible infiltration of any abdominal viscera. The authors present a series of eight patients and emphasize the management specificities of such patients and the possibility of long-term remissions, regardless of the severity of the initial presentation. PMID- 9752481 TI - [Value of surgery in the treatment of advanced cervical cancers]. AB - Surgical resection of advanced nonmetastatic forms of cervical cancers is controversial, but improve local control. The local and regional staging assessment, comprising an examination under general anaesthesia, endocavitary ultrasonography, computed tomography (CT) and/or magnetic resonance imaging (MRI) allows staging, evaluation of the main prognostic factors and selection of the therapeutic strategy. Pelvic and lumboaortic lymph nodes can be investigated by CT, MRI or laparoscopic lymphadenectomy. Surgical resections consist of colpohysterectomy possibly combined with radical lymphadenectomy or pelvic exenteration, followed by pelvic reconstruction using various procedures: low colorectal anastomosis, continent urinary diversion, and vaginal reconstruction with pelvic filling. The mortality and morbidity of pelvic exenteration remain high. It is therefore important to prevent the most frequent complications as effectively as possible. The local control, overall survival and recurrence-free survival can be improved by combining concomitant radiotherapy-chemotherapy and large surgical resection. Some unfavourable local situations can justify palliative pelvic exenteration in highly selected indications designed to improve local control and comfort of survival. PMID- 9752483 TI - [Surgery under hypnosedation. A new therapeutic approach to hyperparathyroidism]. AB - The elective unilateral approach, sometimes under local anaesthesia, offers many advantages in terms of less invasive and faster surgical approach compared to the conventional surgery under general anaesthesia. Nevertheless this approach is restricted to patients unsuspected of multiglandular disease, free from thyroid disease and for whom localization studies are contributive. Surgery under hypnosedation offers the same advantages and provides the possibility of not only exploring the four glands but also of performing a partial thyroidectomy if needed. In our experience 21 patients underwent a cervicotomy under hypnosedation for primary hyperparathyroidism (HPT). No conversion to general anaesthesia was needed; mean operative time was 52 +/- 16 min. In 17 cases, HPT was due to a single adenoma, in 3 cases to hyperplasia (among them a MEN-1 case), and in one last case to a double adenoma. The four glands were identified in 85%. With a follow-up running from 4 to 45 months, all patients are cured. Hypnosedation offers the same medical and economic advantages than the unilateral access under local anaesthesia. Moreover indications are not restricted to selected patients. PMID- 9752484 TI - [Prevalence of thyroid cancer in hot nodules]. AB - The prevalence of thyroid cancer is not nodules is low, as it is estimated to be between 0 and 4% in adults. In order to more accurately estimate this prevalence and to recommend optimal treatment, 93 hot nodules operated between 1976 and 1995 were reviewed. 52 of these subjects (56%) were euthyroid, 11 (12%) suffered from clinical hyperthyroidism and 30 (32%) a presented subclinical hyperthyroidism. All patients were operated for a hot nodule, 1.5 to 6 cm in diameter. Two groups of patients were considered on the basis on the histological examination. Group I consisted of 47 patients (39 F, 8 M, mean age: 44 years +/- 15) with multinodular goitre; 16 (34%) of them underwent total thyroidectomy. Group II was composed of 46 patients (38 F, 8 M, mean age: 44 years +/- 15) with a solitary hot nodule; 39 (85%) patients underwent unilateral lobectomy. Microscopic carcinoma (diameter less tha 1 cm) ws discovered in 2(4%) patients of group 1 and 5 (11%) patients of group 11, corresponding to a total prevalence of 7/93 (7.5%). The microscopic carcinoma was located in the nodule or in the capsule in four cases, and away from the nodule but in the ipsilateral lobe in three cases. In every case, this was an incidental finding and no clinical features distinguished patients with or without microscopic carcinoma. This study suggests that the probability of discovering a microscopic carcinoma associated with a hot nodule is considerable. However, microscopic carcinoma is an incidental discovery in a number of subjects undergoing thyroidectomy and is present in 10 to 20% of autopsied thyroids. As surgical treatment is devoid of risks, it appears indicated, particularly when the nodule exceeds 3 cm in diameter. PMID- 9752485 TI - [Basedow's disease and thyroid nodules. A common association]. AB - The frequency of thyroid cancer associated with Graves' disease is a controversial subject. The authors retrospectively studied 110 consecutive patients operated for Graves' disease between 1991 and 1996. Each patient was evaluated by clinical and laboratory examination, ultrasound and isotope scan. None of the patients had a history of external beam cervical radiotherapy or radioactive iodine. All patients were treated by total or subtotal thyroidectomy. Thyroid nodules were detected in 28 patients (24 women, 4 men), and 6 of them corresponded to papillary carcinoma (5.5% of patients with Graves' disease and 21.4% of patients with Graves' disease associated with nodules). These data suggest the value of surgery in Graves' disease associated with thyroid nodules. PMID- 9752486 TI - [Thyroid carcinoma in a thyroglossal duct cyst: tumor resection alone or a total thyroidectomy?]. AB - Tumours arising in a thyroglossal duct cyst are very rare. Most of them develop from ectopic thyroid remnants. Controversies persist concerning th treatment of these neoplasms, some authors preferring local excision (Sistrunk procedure), while others prefer a more radical approach (associated total thyroidectomy). From 1977 to 1996 we observed and treated 10 patients with by a thyroglossal duct tumour: 8 females and 2 males. A mass in the midline of the neck was the presenting complaint in all cases. Each patient was treated by a Sistrunk procedure associated with total thyroidectomy. Histopathology reports showed 7 papillary carcinomas, 1 Hurthle cell carcinoma, 1 follicular carcinoma and 1 insular carcinoma. Systematic examination of the thyroid gland revealed foci of papillary cancer in 4 cases (40%), with only 1 tumour being larger than 1 centimetre. Cervical metastases were found at operation in 1 case. This series suggests that total thyroidectomy for tumours of thyroglossal cysts could be justified by the high incidence of associated papillary carcinomas of the thyroid and by the relatively aggressive nature that some tumors. In these cases, a radical therapeutic attitude allows, better patients management (total scintigraphy, serum thyroglobulin measurement) and allows the possibility of a complementary radioiodine treatment. PMID- 9752487 TI - [Early therapeutic management of patients genetically predisposed to medullary thyroid cancer]. AB - STUDY: The aim of our study was to study therapeutic results after thyroidectomy in patients positive for predictive genetic analysis and with preoperative calcitonin (CT) response to pentagastlin (Pg) < 150 pg/ml. MATERIAL AND METHODS: 36 patients (13 F, 23 M) were selected: 13 F-MTC from 8 families, 22 MEN 2A from 15 families and 1 MEN 2B. They were positive for direct RET mutation analysis. CT was assayed by immunoradiometric method before and after Pg. Pg test results before and after thyroidectomy, age at operation and histologic results were analysed. RESULTS: Mean preoperative peak CT was 82.5 +/- 34.0 pg/ml (22-133): among these 36 patients preoperative basal and peak CT were normal in 16 and 2 patients respectively. F-MTC and MEN 2A patients were different according to their preoperative peak CT levels (58.1 +/- 24.0 vs 97.6 +/- 31.3) pg/ml, p < 0.01) and age at thyroidectomy (20.4 +/- 10.5 vs 11.6 +/- 7.6 years, p < 0.01 by Mann-Whitney test). Total thyroidectomy was performed in all patients at a mean age of 14.8 +/- 9.8 years (2.5-41.7) and was associated with lymph node dissection in 30 cases. The 2 F-MTC patients with normal preoperative peak CT levels had bilateral C-cell hyperplasia (CCH) associated with uni or bilateral micro-MTC. Other patients had uni or bilateral micro MTC except 4 who had isolated CCH without carcinoma. The age of two MEN-2A and 1 MEN 2B patients with micro-MTC ranged from 2.5 to 4.7 yr. Micro MTC was present in 100% of MEN-2A cases after the age of 10 yr. There were no lymph nodes metastases. During postoperative survey, the last PG tests (n = 33) were performed 27.5 months (1 92) after thyroidectomy: peak CT values were always < 10 pg/ml. IN CONCLUSION: Thyroidectomy should be performed at a very young age in RET mutation carriers, regardless of the plasma CT values. This choice is justified in NEM-2A and NEM-2B patients but must be discussed in F-MTC families with less aggressive forms of the disease. PMID- 9752488 TI - [Microscopic thyroid papillary cancers presenting as cervical lymph node metastases]. AB - Microscopic papillary thyroid cancer (< 1 cm in diameter) is reputed to have an excellent prognosis. In 10 to 20% of cases, it presents in the form of lymph ode metastases. Immediately metastatic forms can be associated with unfavourable prognostic factors, such as multifocal tumours, extension to adjacent tissues, capsular effraction and development of distant metastases. We report 4 cases of microscopic papillary thyroid cancer presenting in the form of lymph node metastases. No primary thyroid lesion was palpable in any of these patients, but the subsequent course was complicated by ling metastases in one case. All patients underwent total thyroidectomy with lymph node dissection followed by adjuvant therapy with radioactive iodine and thyroxin inhibitory treatment. Lung metastases were observed in one case. The authors propose a therapeutic approach based on analysis of severity factors, which determine the recurrence rate. PMID- 9752489 TI - [Death from myocardial metastasis of thyroid cancers (insular, papillary with undifferentiated foyers)]. AB - Two patients, both 70 years old, operated for differentiated thyroid cancer with foci of undifferentiated tumor (one papillary containing pseudosarcomatoid foci, the other with insular pattern containing undifferentiated foci) suddenly died. In both cases, for both of them metastatic involvement of the myocardium and endocardium. These original findings suggest that elderly patients suffering rom well differentiated thyroid cancer, containing undifferentiated foci, therefore suspected to have metastasized, would benefit from non-invasive cardiac work-up (sonogram) in order to plan their multidisciplinary treatment. PMID- 9752490 TI - [Malacoplakia and colonic cancer. Value of the extemporaneous section]. PMID- 9752492 TI - [Endoscopic esophagodiverticulostomy for Zenker's diverticulum]. PMID- 9752491 TI - [Laparoscopic resection of a gastric leiomyoblastoma]. PMID- 9752493 TI - [Varices of the legs]. PMID- 9752494 TI - [Loosening of cemented total hip prostheses. 31 cases]. PMID- 9752495 TI - [What's new in adjuvant therapy of colorectal cancers?]. AB - The indication for adjuvant chemotherapy after macroscopically complete resection of a colonic cancer with lymph node involvement (Dukes C or stage III) is now established. Ongoing studies are designed to define the optimal modalities. A 6 month postoperative treatment based on 5-FU modulated by folinic acid currently represents standard treatment, as conventional treatment with 5-FU and Levamisole for one year is more constraining, potentially more toxic and not more effective. No published study has yet demonstrated any significant benefit of adjuvant chemotherapy for forms B2 or II, essentially because of a lack of statistical power. The place of local treatment and specific immunotherapy is currently being investigated. In the short- or medium-term, the development of new drugs active in the metastatic setting, as well as genetic prognostic factors, should modify current attitudes concerning indications and modalities of adjuvant chemotherapy for colonic cancers. PMID- 9752496 TI - [Heterotopic pancreas. Three cases]. AB - The authors report three cases of heterotopic pancreas discovered surgically in three young men. The heterotopy was located in the biliary tract in one case revealed by choledochal stenosis (case 1), in the stomach discovered at laparotomy for abdominal trauma (case 2) and in the first part of the duodenum discovered during hepatic resection for adenoma (case 3). Management consisted of Whipple's operation in the first case, tumor resection in the second and antroduodenectomy in the last case. Histological examination revealed no malignant transformation. Results were excellent with follow-up 2-years to 4 years. The authors recommend resection with histological examination for all case of heterotopic pancreas discovered at surgery. PMID- 9752497 TI - [Compression of the celiac trunk by the diaphragmatic arcuate ligament during supramesocolonic surgery]. AB - Coeliac stenosis induced by arcuate ligament compression is usually asymptomatic. Current caution is advised during a supramesocolonic surgical procedure in the case of a collateral arterial system due to coeliac stenosis. 11 cases of coeliac stenosis are described. 3 patients underwent duodenopancreatic resection, 8 patients had liver transplantation. 2 patients died after complications due to celiac, stenosis. These cases are described. The authors discuss the diagnostic and therapeutic approach. PMID- 9752498 TI - [Long term results of esophageal epidermoid cancers in complete remission after preoperative chemo-radiotherapy]. AB - INTRODUCTION: Improvements of the results of combined chemoradiotherapy (CRT) in esophageal cancer has led several groups to adopt a non surgical attitude specially in case of complete response (endoscopy +/- biopsy). Few information are available about the follow-up of these patients. We studied long-term results of 35 patients who underwent resection after complete response to preoperative chemoradiotherapy. PATIENTS AND METHODS: 161 patients with resecable carcinoma of the thoracic esophagus have received the same protocol of CRT (cisplatin 80 mg/m2, radiation therapy split course: 37.5 Gy) all patients were followed every for 4 months (no lost of view). RESULTS: Complete response (endoscopy and biopsy) was obtained for 35 patients (21.7%), 19 of them (54%) had pathologic complete response (PCR) (no tumor in the specimen), 16 have microscopic foci of residual tumor (46%). The overall 5-year survival rate was 49.8% for the whole group (median survival 64 months), 70% for the group without tumor in the specimen, 48% for the group with microscopic foci of residual tumor (NS). CONCLUSIONS: One half of the complete response (endoscopy + biopsy) have not a pathologic complete response (microscopic foci of residual tumor in the specimen). The 49.8% of five year survival suggests a benefit from esophagectomy for complete response after combined chemoradiotherapy. PMID- 9752499 TI - [Continuing education. Treatment of hepatocellular carcinoma in cirrhosis. Introduction]. PMID- 9752500 TI - [Epidemiology and diagnosis of hepatocellular carcinomas in cirrhosis]. AB - Hepatocellular carcinoma (HCC) is the most frequent primary cancer of the liver and the most frequent tumour in males, worldwide. The annual incidence of HCC is maximum in Asian and African countries, lower in western countries where it is close to 4/100,000 inhabitants. In 90% of the cases, HCC complicates course of liver cirrhosis, with an annual incidence in cirrhoties of 2 to 6%. Risk factors for HCC in cirrhotics are male gender (sex-ratio: 4/1), age (above 50 years old), macronodular cirrhosis and large cell dysplasia. HCC can complicate the course of cirrhosis of any cause, but might be less frequent in primary biliary cirrhosis, Wilson's disease and auto-immune hepatitis. Currently, the diagnosis of HCC is usually considered in the presence of a focal nodular lesion, during systematic ultrasonographic examination of the liver. In high incidence areas, HCC can still be diagnosed because of HCC-related symptoms. In the case of a focal lesion discovered on a cirrhotic liver, the diagnosis of HCC can be confirmed by studying the behaviour of the lesion of helical CT scan of the liver (enhancement of the tumour during the arterial phase) or MRI (hyperintensity of the tumour on T2 relaxation time); study of peritumour vessels can also be helpful. Serum alpha foeto-protein level, when higher than 300 to 500 micrograms/L is very specific of HCC. When aggressive treatment of HCC is considered and when the diagnosis of HCC remains uncertain, HCC can be assessed by means of cytological or histological study of the tumour on samples taken by fineneedle aspiration (80% sensitivity) or liver biopsy during laparoscopic laparotomy. Forthcoming improvements in imaging technology might eliminate the need for such invasive diagnostic techniques in the future. PMID- 9752502 TI - [Chemoembolization techniques for hepatocellular carcinoma in cirrhosis]. AB - Transcatherter oily chemoembolisation, that should not be confused with other different and less effective techniques also called "chemoembolisation" is the most widely used therapy for loco-regional palliative treatment of hepatocellular carcinoma, which will become increasingly frequent, due to HCV infection; the cancer itself is often discovered at an advanced stage, when neoplastic extension precludes radical treatment that is liver transplantation. Performed with the best techniques, it offers a 1- and 5-yr survival of 60 and 30%, that is under confirmation by a randomized trial. PMID- 9752501 TI - [Value of percutaneous ethanol injection in the treatment of hepatocellular carcinoma in a cirrhotic liver]. AB - Percutaneous ethanol injection treatment, introduced ten years ago as palliative therapy for patients with inoperable hepatocellular carcinoma, can now be used with a curative intent to treat small tumours with results comparable to surgical resection. This progress, made possible by sophisticated radiological techniques, makes percutaneous ethanol injection the treatment of choice for patients with poor liver function in whom resection is not possible and local control of the disease is desirable either for prolonged palliation or in view of liver transplantation. For patients with large tumours, or in case of recurrence after previous surgical treatment, a therapeutic approach combining transarterial lipiodol chemoembolisation and percutaneous ethanol injection has shown promising results and deserves further investigation. PMID- 9752503 TI - [Surgical treatment of hepatocellular carcinoma in cirrhosis]. AB - Despite the recent development of percutaneous ethanol injection and liver transplantation, liver resection remains the reference treatment for hepatocellular carcinoma (HCC). The two drawbacks of this treatment are the risk associated with surgery and the high recurrence rate. Both are related to the almost constant presence of a chronic underlying liver disease. The risk of surgery has decreased significantly over the past 10 years and is currently less than 10%, even after a major hepatectomy, provided that cirrhosis is compensated (Child A) and that there is no superimposed chronic active hepatitis. Recurrence is usually related to de novo carcinogenesis Adjuvant and neoadjuvant therapies have no clearly demonstrated benefit. However, postoperative follow-up is mandatory as some recurrences are arnenable to local treatment, particularly rehepatectomy that has an efficacy comparable to that of first hepatectomy. PMID- 9752505 TI - [Indications and results of liver transplantation in the treatment of hepatocellular carcinoma in cirrhosis]. AB - Liver transplantation is a treatment for hepatocellular carcinoma in cirrhosis which is both recognized, because potentially radical, and controversial because associated with a high risk of recurrence. This study reports the results of a consecutive series of 125 patients transplanted for hepatocellular carcinoma in cirrhosis over an 11-year period. Liver transplantation was indicated because of the tumour in 92 cases (74%) and the tumour was an incidental finding in 13 cases (10%) or was discovered on histological examination of the hepatectomy specimen in 20 cases (16%). The operative mortality at two months was 4% with a 20% morbidity, due to vascular (6%) or biliary (14%) complications. Tumour recurrence was observed in 26 patients (21%) Recurrence was exceptional in the incidental or histological forms of hepatocellular carcinoma (5%) and more frequent when the tumour constituted the indication for transplantation (27%). The risk of recurrence and the survival were significantly influenced by the maximal tumour diameter (greater than 30 mm), the number of tumour nodules (greater than 3) and the presence of portal invasion. Inclusion of these factors in patient selection during the second phase of the study allowed a reduction of the risk of recurrence from 33 to 11% and improvement of the 3-year post-transplantation survival from 53 to 76%. Tumours less than or equal to 30 mm in diameter, with no more than 3 nodules, and without portal invasion are ideal indications for transplantation. Tumours with more than 3 nodules and larger than 30 mm appear to constitute a contraindication to transplantation, unless tumour reduction can be achieved by chemoembolization. Intermediate forms of hepatocellular carcinoma between these two extreme forms are possible indications for transplantation, depending on the availability of liver transplants. PMID- 9752504 TI - [Treatment of hepatic recurrence after resection of hepatocellular carcinomas]. AB - Between October 1990 and December 1995, 86 patients underwent hepatic resection for hepatocellular carcinoma (HCC). All resections were carried out with the aim of achieving complete cure. Fifty one (60%) of these patients subsequently developed recurrent HCC. Only twenty patients could be treated in our hospital. There were 18 men and 2 women, with a mean age of 61 years at the time of recurrence. Six patients had a normal liver. Fourteen patients had associated liver cirrhosis. using Pugh's classification, 7 patients were Pugh A, 6 Pugh B and 1 Pugh C. The initial hepatic resection had consisted of major hepatectomy in 9 cases and segmentectomy in the remaining 11 patients. The mean time to recurrence was 17 months. There were 3 recurrences on the resection margin and 17 recurrences away from the hepatic stump. The therapeutic choice after hepatic recurrence was based on the number of tumors, hepatic function and the size of the liver remnant. Six patients were treated by tamoxifen due to poor hepatic function; median survival after recurrence was 6 months. Four patients with a single recurrent tumor on an atrophied liver remnant were treated by percutaneous ethanol injection with a median survival after recurrence of 15 months. Five patients with multiple diffuse lesions and good hepatic function were treated by transarterial chemoembolisation with a median survival after recurrence of 30 months. Five patients with a solitary tumor and good hepatic function underwent a second hepatic resection with a median survival after recurrence of 35 months. The overall median survival after diagnosis of recurrence was 20 months. These results suggest that an active treatment should be carried out in cases of recurrence of HCC. A second resection, if technically possible, offers the best chance of survival. PMID- 9752506 TI - Progress and prospects in hepatocellular carcinoma surgery. AB - Hepatocellular carcinoma (HCC), the second cause of cancer death in China, is responsible for 130,000 deaths every year. However, as a result of efforts in early detection and small HCC resection, re-resection for subclinical recurrence, cytoreduction and second-stage resection for unresectable HCC, and aggressive palliative surgery other than resection (hepatic artery ligation/cannulation, cryosurgery, etc.), and encouraging improvemental of long-term survival of inpatients has been observed in the authors' institution. In the entire series of 2672 HCC inpatients, the 5-year survival was 4.8% in 1958-70, 12.2% in 1971-83, and 46.7% in 1984-95. The 5-year survival rates were: 61.3% for small HCC resection (n = 645), 33.6% for large HCC resection (n = 950), 48.9% for re resection (n = 147) calculated from the first resection, 67.9% for second-stage resection (n = 73), and 19.8% for palliative surgery (n = 574, including 73 with second-stage resection). By 1995, 239 HCC patients survived for more than 5 years. Recurrence and metastasis remained the major obstacles to more prolonged survival after HCC surgery. Molecular studies of HCC invasiveness, experimental intervention in the newly established highly metastatic model of human HCC in nude mice in the authors' institution have also been delineated. It is concluded that early detection and small HCC resection remain the major approach to improve survival. Aggressive surgical treatment, such as re-resection and second-stage resection, are also important, and invasiveness-related recurrence will be the next target to be studied. PMID- 9752508 TI - [Surgery in De Medicina of Celsus]. AB - Rediscovered during the Renaissance, Celsus' work (1st Century AD) is the first latin encyclopaedia. We only have his medical works, and we are indebted to him for his classification of diseases according to treatment: by diet, by drugs and manually, that is by surgery. Although surgery was described in Hippocrate's works, Celsus gave the first latin presentation, outlining, general and localised operations, in the VIIth book of De Medicina. The VIIIth book is devoted to orthopaedic operations which are sometimes quite different from Hippocrate's descriptions. For the first time, he describes many surgical instruments, most of which have been found by archaeologists. Finally, ethical considerations widen the epistemological field of surgery and separate it as autonomous specialty. PMID- 9752507 TI - [Treatments for hepatocellular carcinoma in cirrhosis : practical aspects]. AB - Many treatments are able to eradicate or slow the progression of hepatocellular carcinoma in cirrhosis. All are described in this issue of Annales de Chirurgie. These various alternative mean that treatment can be adapted to different clinical situations determined by the patient's general state, the size of the liver tumour, its extrahepatic dissemination and the functional quality of the underlying liver parenchyma. Liver hepatic transplantation, offers a good chance of cure, provided it is reserved for patients in good general condition with a small hepatocellular carcinoma confined to the liver. This situation is increasingly frequent. PMID- 9752509 TI - [Difficult colo-colonic or colo-rectal anastomoses: trans-mesenteric anastomoses and anastomoses with right colonic inversion]. PMID- 9752510 TI - [Laparoscopic manual intestinal anastomosis: experimental study in a pig model]. PMID- 9752511 TI - [Omental attachments of the gallbladder]. PMID- 9752512 TI - Surgery: art or science? Birth of organ transplantation. PMID- 9752513 TI - [Surgical treatment of acute dissection of the ascending aorta (20 years experience)]. AB - PURPOSE: In 1977, the use of gelatine-resorcine-formaline (GRF) biological glue during surgery of acute type A aortic dissection was proposed. The present study retrospectively analyses the late results obtained with this adjunct in an experience extending over a 20-year period. PATIENTS AND METHODS: From January 1977 to July 1997, 193 patients (139 males and 54 females) aged from 15 to 79 years (mean age: 53 +/- 14 years) underwent an emergency operation for type A aortic dissection in our institution. All patients suffering from acute type A dissection and 162 (84%) were operated on within 48 hours after the onset of symptoms. Twenty-eight patients (15.2%) had Marfan's syndrome. In all patients the ascending aorta was replaced and the aortic stumps were reinforced with the GRF glue. In 43 patients (22.2%), the aortic valve was replaced either independently (5 cases-2.5%) or by means of a composite graft (35 cases-19.5%). Recently three patients underwent a complete replacement of the ascending aorta and coronary reimplantation with preservation of the native aortic valve. Because of the location of the intimal tear, the aortic replacement was extended to the transverse arch in 58 patients (30%). RESULTS: Hospital mortality amounted to 21% (40 patients) (22.8% in patients with arch replacement and 20.3% in patients without arch replacement) (ns). The survivors were surveyed from 2 months to 20 years post-operatively (cumulative follow-up: 856 pt/years, mean follow-up: 85 +/ 66 months). During this period of time, 23 patients (15%) had to be reoperated on for a total of 29 procedures. Six of those patients (26%) died at reoperation. At univariate analysis, presence of Marfan's syndrome (P < 0.05) and absence of arch replacement (P < 0.02) were determinant risk factors for reoperation. Emergency (P < 0.01) and thoraco-abdominal replacement (P < 0.04) were determinant risk-factors of death at reoperation. The actuarial freedom from reoperation (Kaplan-Meier, CI: 95%) was: 96.5% (90.9-98.2), 87.6% (79.8-92.7), 80.9% (70.8-88.1), 66.4% (51.1-78.9) at one, 5, 10 and 15 years, respectively. A total of 36 patients (27.7%) died during follow-up. Presence of Marfan's syndrome (P < 0.01), reoperation (P < 0.02), stroke (P < 0.05), cardiac failure (P < 0.05) were determinant risk factors of late mortality. The actuarial late survival rate (Kaplan-Meier. CI: 95%), including hospital mortality, was: 71.5% (64.3-77.8), 66% (58.3-73), 56.4% (47.7-64.7), 46.3% (36.4-56.5) at one, 5, 10 and 15 years. CONCLUSION: The GRF glue has proved to be extremely useful during initial emergency surgery for acute type A dissection, making the procedure much easier and safer. Through this operative improvement, the use of the GRF glue seems to have a beneficial influence on the late results which, however, depend mainly on the patient's basic condition. PMID- 9752514 TI - [Identification of axillary sentinel node by lymphotropic dye in breast cancer. Feasibility study apropos of 128 cases]. AB - AIM OF THE STUDY: The goal of this study was to evaluate the technical feasibility of sentinel node biopsy in breast cancer and its predictivity of axillary node status. PATIENTS AND METHODS: Between January 1996 and June 1997, 128 patients with invasive breast carcinomas, referred to the Cancer Center of Strasbourg and Lyon (France), underwent lymphatic mapping (Patent Blue dye) and sentinel node biopsy followed by axillary clearance (Berg's level I to II). RESULTS: Sentinel node was identified in 76.5% of cases and was predictive of axillary status in 94.9% of cases. The false negative rate of the procedure was 5.1%. Sentinel lymph node was involved in 43.9% of cases and it was the only one involved in 30.2% of cases. The sensitivity of the procedure was 94% (CI: 95% = [88%-98%]) and its specificity 100%. CONCLUSION: Actually considered as new attractive procedure under ongoing evaluation in prospective controlled trials, this study confirms the feasibility and reproductibility of lymphatic mapping and sentinel node biopsy, first stage before entering a new era of minimally invasive axillary surgery in breast cancer. PMID- 9752515 TI - [Results of 10 years of a randomized trial of neoadjuvant chemotherapy in breast cancers larger than 3 cm]. AB - AIM OF THE STUDY: The aim of this randomised trial was to determine advantages and drawbacks of neo-adjuvant chemotherapy in patients with operable breast cancers > 3 cm. MATERIAL AND METHODS: Two hundred and seventy-two women (age 70) with operable breast cancers larger than 3 cm (T2-3/N0-1/M0) were included in a randomised trial from January 1, 1985 to April 30, 1989. Patients in group A (n = 138) were treated by mastectomy and axillary node dissection. Adjuvant chemotherapy was indicated for 104 patients with axillary node involvement (n = 82) or negative oestrogen and progesterone receptors (EPR-) (n = 22). Patients in group B (n = 134) were treated by initial chemotherapy (identical as in group A) followed by locoregional treatment according to the response. Before treatment, the average of clinical tumoural diameter was 43 mm. RESULTS: The median follow up was 124 months. In group B, 49 patients (36.5%) were resistant to chemotherapy; a conservative breast surgical treatment was performed in the other 84 patients sensitive to chemotherapy (62.6%). In this last subgroup, 19 (22.6%) needed a secondary mastectomy because of locoregional recurrence. Survival rates were not different in groups A and B, but loco-regional recurrences were frequent in group B. At 10 years, the overall survival rate was 60% and half of living patients in group B were free of cancer and with their breast. CONCLUSION: Neoadjuvant chemotherapy permitted in two-thirds of cases breast conservation treatment, initially considered to be impossible. Locoregional recurrences are more frequent than after mastectomy and adjuvant chemotherapy. PMID- 9752516 TI - [Results of fundoplication by laparoscopic approach in the treatment of gastroesophageal reflux. Apropos of 224 cases]. AB - STUDY AIM: The aim of this paper is to evaluate prospectively immediate and 2 year results of laparoscopic fundoplicature (LF) for gastroesophageal reflux disease (GERD). PATIENTS AND METHODS: Patients presenting GERD who had been previously submitted to a long-term medical treatment were included in this study. Preoperative workup included upper GI tract endoscopy, esophageal manometry and 24-hour pHmetry. Standard surgical procedure incorporated a Nissen Rossetti 360 degrees fundoplicature. Short vessels division (Nissen operation) was performed in case of high strength of the wrap and a partial fundoplicature (Toupet 270 degrees) was performed when motility disorders of the esophagus were demonstrated by manometry. Postoperative morbidity and results were evaluated, with a clinical appreciation at 3 and 22 months, and by manometry and pHmetry at 3 months. RESULTS: Two hundred and thirty-five patients were observed, and 224 included in the study (143 men and 92 women). Nissen-Rossetti fundoplication was performed in 169 cases (80%), Nissen in 30 (14%) and Toupet in 13 (6%). In 12 cases (5%). LF was converted to an open Nissen-Rossetti procedure. There was no hospital mortality and complications were noted in three cases (1.5%): pneumonia (n = 2) and gastroplegia (n = 1). With a mean 22-month follow up, among the 103 patients who answered to a questionnaire, the rate of relapse of GERD was 14%, dysphagia was present in 2% and four patients had been reoperated on (one for a slipped Nissen, one for a stenosis of the esogastric junction and two incisional hernias). CONCLUSION: On the basis of this experience, LF for GERD is a safe and efficient operation, with 86% of good results at 2 years. PMID- 9752517 TI - [Laparoscopic appendectomy in the adult]. AB - AIM OF THE STUDY: The benefits of laparoscopic appendectomy remain controversial. The aim of the study was to evaluate the advantages and disadvantages of this technique. MATERIAL AND METHODS: Four hundred and forty-eight patients operated on for suspected appendicitis during a 5-year period were analysed in a retrospective study. The preoperative diagnosis was corrected in 21.4% of the cases (8.1% for males and 28.7% for females) and the conversion rate was 9.7%. There were 17 patients with generalized peritonitis and 28 with localized peritonitis. During the past year, this surgical method was introduced in another hospital and a prospective study included 92 consecutive patients operated on for appendicitis. The operating time was 53 minutes and the conversion rate was 7.6%. There were five patients with generalized peritonitis and eight with localized peritonitis. RESULTS: There were no postoperative deaths. In the first period, the morbidity rate was 2.3% in the laparoscopic group without conversion. After pathological examination, there was a 14.2% rate of normal appendix. The mean hospital stay was 4.3 days. In the second period, there were only three minor complications and the mean hospital stay was 4.19 days. CONCLUSION: The choice of laparoscopic approach is associated with some advantages: corrections of the diagnosis (mainly in young women) and simplification of the postoperative course, provided that the surgeon has sufficient experience. PMID- 9752518 TI - [Evaluation of pancreatic cancer by combined laparoscopy and echolaparoscopy]. AB - STUDY AIM: This prospective study was undertaken to evaluate the efficiency of staging laparoscopy associated with laparoscopic ultrasonography in the assessment of tumoural extension and surgical resectability in patients with carcinoma of the pancreatic head. PATIENTS AND METHODS: From June 1995 to March 1997, 26 consecutive patients (11 male and 15 female patients), with a mean age of 62.5 years, were included in this study. The lesion was located in the pancreatic head with jaundice. Four staging methods were used: percutaneous ultrasonography (n = 26) computed tomography (n = 26), endoscopic ultrasonography (n = 26). The assessment of resectability by each procedure was verified by surgical exploration and histologic examination. RESULTS: Results of percutaneous ultrasonography and computed tomography were similar, predicting unresectability in 50% of the patients. Endoscopic ultrasonography performed in the 16 patients without visible metastases according to the previous procedures predicted surgical resectability in seven patients only. With staging laparoscopy associated with laparoscopic ultrasonography, undiscovered metastases were found and unresectability was predicted in 21 patients out of 26; the sensitivity was 100% for liver metastases, peritoneal metastases and vascular involvement, 90% for lymph node involvement and 88% for diagnosis of the primitive lesion. A Whipple procedure was performed in five patients and a palliative bypass in all the other patients except one. An unnecessary laparotomy was avoided in 12 patients. CONCLUSIONS: Staging laparoscopy associated with laparoscopic ultrasonography is superior to all other staging methods. It should be the first step of a potentially curative surgical treatment (five cases only in this series) or of a palliative bypass. Laparotomy was avoided in 12 cases. PMID- 9752519 TI - [Adrenal metastases of hepatocellular carcinoma. Therapeutic options]. AB - AIM OF THE STUDY: The aim of this study was to evaluate the therapeutic modalities, particularly surgical treatment, for adrenal metastasis of hepatocellular carcinomas. PATIENTS AND METHODS: This series included 13 patients (mean age: 64 years) with hepatocellular carcinoma on cirrhotic liver (n = 8) or healthy liver (n = 5). The adrenal metastasis was synchronous in four cases, metachronous in nine, unilateral in ten cases, and bilateral in three. Resection of the adrenal metastasis was performed in seven patients, combined with the resection of the hepatic carcinoma in three cases or secondarily performed in four. The metastasis was not resected in six patients because of poor liver function or poor patient conditions; two patients were treated with percutaneous ethanol injection, one with radiation and three received only a symptomatic treatment. RESULTS: After adrenalectomy combined with liver resection, two patients died in the postoperative course in relation with pulmonary embolism (n = 1) or acute pancreatitis (n = 1). The mean survival in the five other patients was 38 months after the adrenalectomy and 58.6 months after the liver resection. After percutaneous ethanol injection, one patient survived 47 months and the other one 7 months only. After radiation, the patient survived 18 months. After symptomatic treatment, the mean survival was only 7.3 months. CONCLUSION: The present data suggest that adrenal metastasis, either isolated or associated with a well-controlled intrahepatic recurrence, could be treated surgically when the resection is technically feasible. This aggressive management seems the only chance to offer a long-term survival to selected patients. PMID- 9752520 TI - [Treatment of iliac artery stenosis and obliteration by transluminal angioplasty]. AB - PURPOSE: Percutaneous transluminal angioplasty (PTA) has an established and valuable role in the treatment of iliac stenoses and occlusions. The use of stents may improve the results of PTA. The aim of this study is to report our surgical experience of iliac angioplasty over the past 5 years. PATIENTS AND METHODS: From January 1993 to October 1997, 201 iliac PTA were performed in 175 patients, at the level of the common iliac artery (n = 111) and external iliac artery (n = 90). In 26 cases, two iliac lesions were treated simultaneously. There were 188 stenoses and 13 chronic occlusions. During PTA, 55 stents were used, because of unsatisfactory results (dissection, residual stenosis) or in case of total occlusion, in the common iliac artery in 35 cases and in the external iliac artery in 20 cases. In 15 cases (8.6%), a femoro-popliteal bypass was associated to the iliac PTA, because of multilevel occlusive disease. RESULTS: There were no early deaths. There were seven initial failures. The initial success rate was 96%. The clinical follow-up has been achieved in 163 patients, (range: 3 to 48 months, mean: 28 months). The primary patency rates were 84%, 79% and 63% respectively at 1, 2, and 4 years. The secondary patency rates were 87%, 85% and 73% respectively at 1, 2, and 4 years. PMID- 9752521 TI - [Prosthetic replacement of the ureters]. AB - STUDY AIM: The aim of this study is to demonstrate the reliability of silicone prosthesis for the replacement of ureters. This prosthesis derives from the biliary prosthesis developed after a personal experimental study continued by Triboulet. PATIENTS AND METHODS: In 38 patients suffering from a malignant disease, a right silicone prosthesis was used for the replacement of an ureter during a 20-year period. There were 30 female and eight male patients. The mean age was 71 (range: 51-88 years). Forty one prostheses were used; one patient underwent two successive operations on the same side with a change of prosthesis, and two patients required a bilateral prosthesis. There were 12 gynaecological carcinomas (three with ureteral fistula), three prostatic carcinomas, 16 cancers of the rectum and recto-sigmoid junction, four cancers of the right colon with retroperitoneal carcinomatosis, and three ureteral fistulas after extended colonic resection. RESULTS: Early complications were limited to ureteral fistulas (n = 6, 16%) in patients who had already a preoperative fistula (n = 3) and in patients with peritoneal metastases on the superior wall of the bladder. The secondary destruction of the kidney (four secondary nephrectomies) occurred when the function of the kidney was already impaired at the time of the procedure. There were no secondary fistulas, no secondary obstruction of the prosthesis. The longest follow-up was 69 months. CONCLUSION: The silicone prostheses used for the replacement of ureters are reliable and still permeable beyond 5 years. The protection of the renal function in patients often submitted to chemotherapy improves the duration and quality of survival. These prostheses must be reserved to advanced malignant diseases with a rather long life expectancy. PMID- 9752522 TI - [Presacral myelolipoma]. AB - Presacral myelolipoma is a rare benign tumour of unknown aetiology, composed of mature adipose tissue with intermixed normal haematopoietic cells. Computed tomography is of help in the diagnosis but biopsy is mandatory in order to avoid unnecessary surgery. A case is reported. PMID- 9752523 TI - [Diaphragmatic or juxtadiaphragmatic schwannoma?]. AB - The authors report one case of schwannoma of diaphragmatic topography. This lesion is exceptional and, to our knowledge, only seven cases have been published in the literature. The first diagnosis was a suprarenal tumour. Magnetic resonance imaging provided more detail about the juxta diaphragmatic topography of the tumour, which was subsequently confirmed by surgery. The diagnosis of benign schwannoma was made at the pathological and immuno-histochemical examination of the specimen. The schwannoma corresponded to the type B of the classification of Antoni. Pathogeny and origin of the tumour are discussed. The sympathetic nervous para-vertebral system, the phrenic or intercostal nerves could be the origin of the tumour. In the eight cases reported, the tumoural removal was performed through thoracotomy (n = 5) or laparotomy (n = 3). The preoperative exact location of the juxta diaphragmatic tumours remains difficult to specify. PMID- 9752524 TI - [Contributions of genetics in gastroenterology. Has genetic progress changed the therapeutic management of colorectal neoplasms? "Colon" Group of the Curie Institute]. PMID- 9752525 TI - [Virtual colonoscopy with computerized tomography]. PMID- 9752526 TI - [Baron Percy (1754-1825). Chief surgeon of the Grande-Armee]. PMID- 9752527 TI - [Survival of the patient operated for cancer of the rectum as a function of the experience of the surgeon]. PMID- 9752528 TI - [Surveillance after curative resection for colorectal cancer. Prospective controlled study]. PMID- 9752530 TI - [Laparoscopic versus Young-Mayor open posterior adrenalectomy: a case-control study of 100 patients]. PMID- 9752529 TI - [Reinsertion of the distal common bile duct into the resection cavity during duodenum preserving resection of the head of the pancreas for chronic pancreatitis]. PMID- 9752532 TI - [Eulogy for Gilles Edelmann (1917-1996)]. PMID- 9752531 TI - [Videolaparoscopic digestive system surgery: 10 years already!]. PMID- 9752533 TI - [Prevention of intestinal allograft rejection by anti-adhesion molecule antibodies in a mouse model]. AB - STUDY AIM: Small bowel transplantation is still hampered by a high morbidity and mortality linked to the heavy non specific immunosuppression which is required by the transplantation of this lymphoid organ. Adhesion molecules appear to be potential targets for specific immunosuppression. The aim of the study was to investigate the effect of a transitory administration of anti-LFA-1 or anti-alpha 4 monoclonal antibodies (mAb) in the prevention of rejection in a model of fetal small-bowel transplantation in mice. MATERIALS AND METHODS: The small bowel of C57BL/6 (H-2b) fetus (16 to 20 days of gestation) was transplanted into adult C3H/He mice (H-2k) or C57BL/6 recipient mice. Recipients were treated with a short course of either anti-LFA-1 mAb alone, either with anti-alpha 4 mAb alone, or with both mAb. Biopsies with histological study of the grafts were performed between post-operative day 5 and 60. A score of development and rejection was assigned to each sample. RESULTS: Normal intestinal development with no sign of rejection was observed in 24/28 syngenic grafts till post-operative day 45. In the absence of treatment, intense rejection was observed as soon as day 5 and all allogenic grafts were rejected (n = 22). In contrast, in anti-LFA-1 mAb treated mice, 18/20 allogenic grafts developed normally with minimal signs of rejection. In anti-alpha 4 treated mice, a transient protective effect on small bowel allograft survival was observed on day 7 but thereafter, all grafts were massively rejected within a few days (n = 18). The combination of both mAb didn't improve the survival of the grafts when compared to anti-LFA-1 mAb treated grafts (n = 10). CONCLUSION: These results demonstrate that a transitory administration of anti-LFA-1 mAb, but not of anti-alpha 4 mAb, is able to prolong significantly the survival of non vascularized small bowel fetal grafts in mice. Our results are promising for the possible use of the anti-LFA-1 mAb in clinical intestinal transplantation. PMID- 9752534 TI - [What place for endoscopic sphincterotomy in treatment of acute pancreatitis?]. AB - STUDY AIM: A prospective study was undertaken in order to evaluate the effects of endoscopic sphincterotomy on the evolution of biliary and idiopathic acute pancreatitis. PATIENTS AND METHODS: Among 320 patients with acute pancreatitis observed from 1986 to 1996, 118 were excluded from the study for etiological reasons and 137 were included for an endoscopic sphincterotomy within 72 hours from their admission. There were nine technical failures and 128 endoscopic sphincterotomies were performed. Sixty-five eligible patients were not included for logistic problems or patients' refusal; they can be considered as a "control group". RESULTS: The mortality rate of endoscopic sphincterotomy was 0 and the morbidity rate 2.1%. The mortality rate of acute pancreatitis was 3.1% in the sphincterotomy group vs 7.6% in the control group (P = 0.1) (NS) and the morbidity rate 25% versus 32% (P > or = 0.1) (NS). CONCLUSION: These results suggest that endoscopic sphincterotomy could be beneficial in acute biliary or idiopathic pancreatitis but they are not statistically significant. Endoscopic sphincterotomy does not increase the severity of acute pancreatitis and can be considered particularly in cases of gallstone pancreatitis but it should be performed less than 48 hours after the onset of acute pancreatitis. PMID- 9752536 TI - [Superficial stomach cancers: surgical experience and study of prognostic factors in 102 patients]. AB - STUDY AIM: The aim of this retrospective study was to analyze the characteristics, treatment and prognosis of early gastric carcinoma in a series of 102 patients. METHODS: Between 1973 and 1994, 102 patients (68 males, 34 females) with a mean age of 65 years, were operated on for an early gastric carcinoma. Mean follow-up was 7 years. Survival was calculated using the Kaplan Meier method. Prognosis was determined with univariate and multivariate analysis according to Cox model. RESULTS: The carcinoma was limited to the mucosa in 57 patients (56%) and extended to the submucosa in 45 (44%). There was a lymph node invasion in 17 patients (16.5%). The postoperative mortality rate was 5.8% (n = 6). Secondary deaths occurred in relation with the gastric cancer in 10.4% (n = 10). The 5- and 10-year actuarial crude survival rates were 84% and 68.6%, respectively. Univariate analysis of prognosis factors showed a significant survival difference according to the age (P = 0.001), submucosal extension (P = 0.03), lymph node invasion (P = 0.0005) and type of gastric resection performed (P = 0.03). With multivariate analysis of prognostic factors, advanced patient age and lymph node metastases were the only statistically significant independent prognostic factors, advanced patient age and lymph node metastases were the only statistically significant independent prognostic factors (P = 0.0002 and P = 0.002, respectively). CONCLUSIONS: Prognosis of early gastric cancer is usually excellent. Patients with high risk of recurrence may be identified in relation with prognostic factors and mainly with lymph node invasion. PMID- 9752535 TI - [Gastrointestinal hemorrhage caused by rupture of an aneurysm of visceral arteries. Presentation of 4 cases]. AB - STUDY AIM: Gastrointestinal bleeding by rupture of splanchnic artery aneurysms is very rare. The aim of this study is to report four cases observed between 1990 and 1996. MATERIALS AND METHODS: In the first case, the celiac trunk aneurysm was revealed by hematemesis due to erosion of the posterior wall of the stomach. Excision of the aneurysm associated with splenopancreatectomy was followed by revascularization of the common hepatic artery with a bypass implanted in the aorta. The second case concerned a splenic artery aneurysm revealed by hemosuccus pancreaticus and intestinal bleeding which was treated by excision and splenopancreatectomy. In the third case, the common hepatic artery aneurysm revealed by hemosuccus pancreaticus and intestinal bleeding was treated by obstructive endoaneurysmorrhaphy. The fourth case concerned a superior mesenteric aneurysm revealed by duodenal erosion causing gastric and intestinal bleeding, which was treated by obstructive endoaneurysmorrhaphy and revascularization of the mesenteric artery by a spleno-mesenteric bypass. RESULTS: Surgical treatment was successful in all four patients. In the first case, an acute acalculous cholecystitis required a cholecystectomy after 3 weeks. In the fourth case, a splenic infarction disappeared spontaneously. CONCLUSION: Such observations are rare. The site of the bleeding was located by endoscopy. The aneurysm was recognized by contrast-enhanced computerized tomography (CT) scan and/or celiac and mesenteric arteriography which was performed in all cases and was very useful for the management of such aneurysms. After excision (n = 2) or obliterative endoaneurysmorrhaphy (n = 2), revascularization had to be done in two cases for celiac and mesenteric aneurysms. PMID- 9752537 TI - [Postoperative incisional hernias: intra- or extraperitoneal prosthesis implantation?]. AB - STUDY AIM: The aim of this retrospective study concerning the repair of postoperative incisional hernia using Dacron mesh was to compare results according to the extra- or intraperitoneal mesh position in order to assess the respective indications of each option. MATERIALS AND METHODS: From January 1985 to December 1996, 172 patients (mean age: 61.3 years) were operated on using Dacron mesh extraperitoneally (n = 99) or intraperitoneally located (n = 73). For statistical analysis, both groups were compared using Chi square test or Fisher's test. RESULTS: There were no postoperative deaths in the group with extraperitoneal mesh and two postoperative deaths in the group with intraperitoneal mesh. There were no significant differences when results comparing parietal complications (sepsis: 2% vs 2.7%, pain: 9.1% vs 16.9%), secondary intestinal disorders (2% vs 4.2%) and recurrence rate (4% vs 5.6%) were assessed between extraperitoneal and intraperitoneal mesh. Recurrences were related to pareital infection treated by partial removal of the mesh (n = 2) or to the lateral detachment of the mesh (n = 6). CONCLUSIONS: In the group of patients receiving extraperitoneal mesh there were no postoperative deaths and morbidity was low (this technique is generally used in the treatment of large incisional hernia). In the group of patients receiving intraperitoneal mesh, similar parietal and general complications were observed. But the risk of serious complications and postoperative death is higher; this technique must be limited to the most serious incisional hernia and to high risk patients. PMID- 9752538 TI - [Long-term outcome of infra-inguinal endovascular surgery for critical ischemia]. AB - STUDY AIM: Endovascular surgery can be proposed as an alternative to infrainguinal conventional surgery in critical ischemia. The aim of this study was to report the latest results of our series of 186 patients. MATERIALS AND METHODS: One hundred and eighty-six patients (100 women and 86 men; mean age 74.5 +/- 13 years) were treated for pain during rest (31.5%), gangrene (58%), or ischemic ulcer (10.5%). The lesions were unilateral (n = 172) or bilateral (n = 14). Two hundred eighty-seven target lesions were treated: for stenosis (n = 168) or occlusion (n = 119): of superficial femoral artery (31.7%), popliteal artery (40%) or tibial arteries (28.3%). RESULTS: Technical success was achieved in 81% (15% amputations). The in-hospital mortality rate was 6.5%. The cumulative patency rate was 61 +/- 3% at 12 months, and 52 +/- 6% at 48 months. The limb salvage rate was 87 +/- 3% at 12 months and 82 +/- 4% at 48 months. Thirteen potential factors of patency were analyzed: the only predictive factors affecting patency were occlusion versus stenosis, and the use of atherectomy (Log rank test: P < 0.001 and P < 0.0001). CONCLUSION: Despite a risk of technical failure and of mid-term restenosis, endovascular surgery for critical ischemia provides a fair long-term limb salvage rate. PMID- 9752539 TI - [Gene therapy of cerebral glioblastoma by adenovirus vector. Experimental model in the rat]. AB - Our aim was to test the therapeutic effects of adenovirus-mediated gene therapy in an animal brain tumor model which was obtained by stereotactic injection of 9L gliosarcoma cells into the caudate nucleus of rat brains. Seven days after the implantation of tumor cells, adenovirus vectors bearing the Escherichia coli beta galactosidase gene (ADVbgal) or the herpes simplex virus thymidine kinase gene (ADVtk) were stereotactically injected into the tumor. Injection of the ADVbgal resulted in the expression of the marker gene in 11 animals. Transfer of the ADVtk was followed, 3 days later, by intraperitoneal injection of ganciclovir (GCV) for 10 days. A control group was treated with saline instead of GCV. We observed a significant regression of the tumors in the rats treated with ADVtk and GCV as compared with control animals. In four cases the tumor completely disappeared after treatment. These results demonstrate the potential efficacy of adenovirus-mediated transfer of the HSVtk gene following by GCV administration for the treatment of glioblastomas. PMID- 9752540 TI - [Evaluating the risk of axillary lymph node involvement in inferior breast cancer measuring 3 centimeters. Analysis of a predictive model based on 893 cases]. AB - STUDY AIM: The aim of the study was to assess, by clinical and histological predictive factors, the axillary lymph-node involvement (pN+) in early breast cancers. MATERIALS AND METHODS: Eight hundred ninety-three patients with unilateral invasive breast cancer were studied. The evaluated parameters included clinical size (T), pathological size (pT), histological subtype (ductal infiltrating, according to grading 1, 2, 3, lobular infiltrating and others), age (less than 40, 40 to 60 and above 60). Furthermore, a new parameter, the dosimetric breast size, recently described, was included (Eur J Cancer 1997; 33: 2432-4). RESULTS: The global rate of pN+ was 25.3%, with respectively, pN1: 10%, pN2-3: 8.4% and pN > 3: 6.9%. According to T, the pN+ rates were, respectively, 13.8%, 19.8% and 36.2% in the T0, T1 and T2 < or = 3 cm groups. According to pT, the pN+ rates were, respectively, 11.1%, 17.7%, 23.5%, 30.1% and 36% in the following groups: 0-9.9 mm, 10-14.9 mm, 15-19.9 mm, 20-24.9 mm and 25-29.9 mm. For the ductal infiltrating carcinoma, according to the gradings 1, 2 and 3, we found, respectively, 18.3%, 27.2% and 37.8% of pN+. For the lobular infiltrating carcinoma and the other histological subtypes, the rates were 22.7% and 10%, respectively. For the three age categories cited above the pN+ rates were, respectively, 30.3%, 25.8% and 22.4%. According to breast size we found 30.1% and 24.4% of pN+ respectively for small and medium or large dosimetric breast size. After a multivariate analysis, three factors were significant for pN+ risk: clinical tumor size (P = 0.0001), histological subtype (P = 0.0005) and dosimetric breast size (P = 0.004). With a combination of these three factors, the pN+ rates varied from 5% to 50%. CONCLUSIONS: The authors conclude that both clinical and pathological characteristics of the primary tumor (specified by previous core biopsy) can indicate the risk for axillary node metastases, and allow selection of candidates for limited axilla surgery (sentinel node). PMID- 9752541 TI - [Joint denervation, a simple response to complex problems in hand surgery]. AB - STUDY AIM: In patients with preserved mobility and stability a painful joint remains a difficult problem, especially in elderly patients. All operations, including intracarpal arthrodesis, reduce an already limited mobility, require prolonged immobilization and have a high rate of complications. Denervation could be proposed in such cases. MATERIALS AND METHODS: In our study denervation was performed on 132 wrists, 36 first carpo-metacarpal joints and 32 proximal inter phalangeal joints. RESULTS: We have been disappointed in the past by partial wrist denervations. Fifty cases of complete and isolated wrist denervation were reviewed with a mean 5-year followup. Strength and mobility were only marginally improved but pain was decreased by a mean 75% (on a visual analog scale) in 74% of patients. At the proximal inter-phalangeal joint level, the mean pain improvement was 88% in 85% of patients. At the first carpo-metacarpal joint level, results of denervation were less predictable and the mean pain improvement was 67% in 81% of patients, with a mean 17-month follow-up. CONCLUSION: Joint denervation is a simple but precise operation performed under local anesthesia and on an outpatient basis. It provides good results in elderly patients, with few complications. PMID- 9752542 TI - [Hand replantation: long-term functional results (apropos of 8 cases)]. AB - STUDY AIM: The aim of this study was to report the long term functional results after replantation of the hand in eight patients. MATERIALS AND METHODS: Between 1977 and 1995, hand replantation was performed in eight cases (six males and two females). Mean age at the time of injury was 31 years (24-47 years). The dominant hand was amputated in half of the cases. In two cases, the soft tissue lesions were severe; in the six other cases, the wound was clean-cut. The level of amputation was transmetacarpal (n = 2), carpal (n = 2), wrist (n = 2), distal part of the forearm (n = 2). RESULTS: The mean convalescence time was 16 months (from 6 months to 2 years). The degree of disability ranged from 40 to 65%. Patient follow-up lasted 2 to 20 years (mean: 11 years). The return of discriminative sensitivity of the digits was noted in six cases. The active motion of the fingers was satisfactory in all cases, but intrinsic muscle function was weak or absent. Pinch and grasp strength was reduced from 10 to 60% (when compared to the non-damaged hand). One patient resumed his prior occupation, and another resumed his occupation part-time. The six other patients found new occupations. CONCLUSION: All the patients achieved a useful function of their replanted hand in their daily, spare-time and professional activities. PMID- 9752543 TI - [Acute acalculous cholecystitis due to Taenia saginata]. PMID- 9752544 TI - [Consensus conference. Prevention, detection and management of colon cancer. Brief report]. PMID- 9752546 TI - ["La Grande Chirurgie" by Guy de Chauliac restored by Laurent Joubert]. PMID- 9752545 TI - [Unpredictable risk and damage compensation in civil process]. AB - UNPREDICTABLE RISK DEFINITION: Any medical or surgical action with diagnostic or therapeutic purpose involves a potential risk of complication, unrelated to the initial pathology or a technical error, but could lead to death or disability. Proof of unpredictable risk without initial fault requires a careful examination of each case by a judicial expert in order to eliminate the complications related to an unskilled surgeon or to poor postoperative care. UNPREDICTABLE RISK COMPENSATION: Conclusions of the Academy: In current French common law, damage compensation can only be obtained in cases where all possible medical means are not implemented and professional fault can be proven. It is illogical to extend the civil responsibility of the practitioner to an obligation of procedural result. In the absence of fault, damage compensation does not come under the civil responsibility of the surgeon. A special insurance system for unpredictable risk has to be created using the principles of mutual insurance. PMID- 9752547 TI - [Preoperative radiotherapy and chemotherapy or not in epidermoid cancer of the esophagus?]. PMID- 9752548 TI - [Treatment of common bile duct calculi: laparoscopy or sphincterotomy? Controlled prospective study]. PMID- 9752549 TI - [Dorsal sector of the liver]. AB - The dorsal sector extends in front and to the sides of the inferior vena cava, separating the caval axis from the main liver (excepting superiorly the entrance of the main hepatic veins into the vena cava). The two elements, dorsal sector and retro-hepatic portion of the vena cava, actually make a single unit. It is made of two segments: left (segment I) larger than the Spieghel lobe, right (segment IX) incorporated in the posterior surface of the right liver. The "caudate process" is not a peculiar element: it is nothing else than the inferior margin of segment IX: the breadth gives information on the size of segment IX. The dorsal sector is the midportion of the posterior liver, it is absolutely independent of the right and left livers separated by the main portal fissure. Portal pedicles are numerous and ascendant, they arise from the posterior margin of the transverse portal arch (from right to left: segment VII vein, right lateral vein, right portal vein, left portal vein, segment II vein). The size of the dorsal sector is variable, and can be appreciated by an antero-posterior index. A voluminous sector may be a problem for the surgeon. Segment IX can be divided in three subsegments: IXb under the interval between the right superior hepatic vein and the middle hepatic vein (longer branches can ascend and supply a small portion of the upper surface in front of the vena cava), IXc under the very broad right superior vein, and posteriorly IXd, linked to segment VII. Only segment I and subsegment IXb receive branches from the right and from the left livers. Hepatic veins enter directly the caval axis, some enter the main hepatic veins. The dorsal sector is a large anastomosis between efferent veins and the vena cava. Anteriorly segment I is in contact with segment IV but also with segment VIII, subsegments IXb and IXc with segment VIII and IXd with segment VII. The fissural limit is difficult to locate. Posteriorly division of the triangular and coronary ligaments, section of the dorsal hepatic veins, the right middle and inferior veins allow separation of the liver from the posterior abdominal wall and the inferior vena cava, so the surgeon can reach the dorsal sector. A remarkable error has been commited when the main hepatectomies were described: the dorsal sector was not known and the caudate lobe was considered as a part of the left liver. Actually the dorsal liver is a separate entity covering the inferior vena cava which has no connexion with the main liver; when the main portal fissure is opened up to the anterior surface of the vena cava, the dorsal sector is opened vertically. Interruption of the pedicles must also be considered. For example, in a left hepatectomy, the left portal pedicle is divided, all the left branches for subsegment IXb (which will be preserved) are interrupted; but the left branches from the right portal pedicle are not interrupted and will bleed when the dorsal sector is divided. When splitting the liver for transplantation, some difficulties can occur, especially with the right transplant. A main practical interest is the possible propagation to the dorsal ducts of hilar carcinoma. PMID- 9752550 TI - [A new concept in surgery of the digestive tract: surgical procedure assisted by computer, from virtual reality to telemanipulation]. AB - Surgical simulation increasingly appears to be an essential aspect of tomorrow's surgery. The development of a hepatic surgery simulator is an advanced concept calling for a new writing system which will transform the medical world: virtual reality. Virtual reality extends the perception of our five senses by representing more than the real state of things by the means of computer sciences and robotics. It consists of three concepts: immersion, navigation and interaction. Three reasons have led us to develop this simulator: the first is to provide the surgeon with a comprehensive visualisation of the organ. The second reasons is to allow for planning and surgical simulation that could be compared with the detailed flight-plan for a commercial jet pilot. The third lies in the fact that virtual reality is an integrated part of the concept of computer assisted surgical procedure. The project consists of a sophisticated simulator which must include five requirements: a) visual fidelity, b) interactivity, c) physical properties, d) physiological properties, e) sensory input and output. In this report we describe how to obtain a realistic 3D model of the liver from bi dimensional 2D medical images for anatomical and surgical training. The introduction of a tumor and the consequent planning and virtual resection is also described, as are force feedback and real-time interaction. PMID- 9752551 TI - [Robotics in neurosurgery: current status and future prospects]. AB - Neurosurgery is in essence a field of application development for robots, based on multimodal image guidance. Specific motorized tools have already been developed and routinely applied in stereotaxy to position a probe holder or in conventional neurosurgery to hold a microscope oriented towards a given target. The potentialities of these approaches have triggered industrial developments which are now commercially available. These systems use databases, primarily coming from multimodal numerical images from X-ray radiology to magnetic resonance imaging. These spatially encoded data are transferred through digital networks to workstations where images can be processed and surgical procedures are pre-planned, then transferred to the robotic systems to which they are connected. We have been using a stereotaxic robot since 1989 and a microscope robot since 1995 in various surgical routine procedures. The future of these applications rely mainly on the technical progress in informatics, about image recognition to adapt the pre-planning to the actual surgical situation, to correct brain shifts (for instance), about image fusion, integrated knowledge such as brain atlases, as well as virtual reality. The future developments, covering surgical procedure, research and teaching, are sure to be far beyond our wildest expectations. PMID- 9752552 TI - [Pulmonary thromboendarterectomy with video-angioscopy and circulatory arrest: an alternative to cardiopulmonary transplantation and post-embolism pulmonary artery hypertension]. AB - The best predictor of poor or suboptimum outcome from pulmonary thromboendarterectomy (PTE) is insufficient relief of obstruction, especially in the lower lobes. The aim of this study is to emphasize that the use of video assisted angioscopy may increase the quality of PTE and thus improve outcome. PTE included a median sternotomy, intrapericardial dissection limited to the superior vena cava, institution of cardiopulmonary bypass, deep hypothermia and sequential circulatory arrest periods. PTE was always bilateral and performed through two separate arteriotomies of both main intrapericardial pulmonary arteries. A rigid 5 mm angioscope connected to a video camera was introduced through the arteriotomy into the lumen to increase the visibility and perform the video assisted endarterectomies of all obstructed segmental branches, including normally inaccessible anterior segmental branches. Between January 1996 and December 1997, 48 patients with severe postembolic pulmonary hypertension had PTE. Patients were in New York Heart Association (NYHA) class II (n = 2), III (n = 28) or IV (n = 18) with the following hemodynamics: mean pulmonary arterial pressure (PAP) 53 +/- 13 mmHg, cardiac index 2.16 +/- 0.5 L/min/m2, pulmonary vascular resistances (PVR): 1,152 +/- 414 dyne.s-1.cm-5. Six patients died from alveolar hemorrhage (n = 1), high residual pulmonary pressure and rethrombosis (n = 4) and hypoxic cardiac arrest (n = 1). The functional outcome in surviving patients was as follows: (NYHA) class I (n = 24), II (n = 16) or III (n = 2) with improved hemodynamics: mean pulmonary arterial pressure: 30 +/- 9 mmHg, cardiac index: 2.78 +/- 0.5 L/min/m2, pulmonary vascular resistances (PVR): 484 +/- 159 dynes.s-1.cm-5. Video-assisted angioscopy allows much improved quality and degree of pulmonary endarterectomy. This expands the indications to include patients with previously inaccessible distal disease and candidates for heart-lung transplantation. PMID- 9752553 TI - [Intraperitoneal transplantation of isolated hepatocytes of the pig: the implantable bioartificial liver]. AB - The general aim is to prepare a bioartificial liver to treat acute hepatic failure using allo- and xenogeneic hepatocytes, immunoprotected by macroencapsulation and transplanted into the peritoneal cavity. The goal of this study was to prepare a large amount of isolated porcine hepatocytes, to encapsulate them within biocompatible membranes for transplant in allo- and xenogeneic combinations and to examine the viability and functionality of the cells 6 weeks later. Hepatocyte isolation was performed in 12 kg pigs (n = 15) by dissociation of the liver with collagenase D (1 g) without oxygenation. Encapsulation of the hepatocyte suspension (10(7)/mL) was performed in hydrogel membranes AN69; hollow fibers (2 m x 0.8 mm) and flaskes (1.8 cm), and transplanted to Yucatan pigs (n = 4) and Lewis rats (n = 12). Six weeks later, they were removed to study the cell viability by histological examination, and the production of albumin by immunonephelometry. The rate of isolated hepatocytes was 38 +/- 5 x 10(9)/mL by liver of pig and the mean viability was 93 +/- 2%. Six weeks after transplantation, hepatocytes were viable, organized in lobules, and showed conserved albumin production. The same results were observed for allogenic and xenogeneic combinations. In conclusion, this method of liver dissociation allowed for preparation of a large amount of isolated hepatocytes from a single pig liver, theoretically sufficient to treat a patient with acute liver failure. Hydrogel membranes were well tolerated and allowed immunoprotection without immunosuppression. Transplanted hepatocytes remained functional. This work is an important step in progress toward clinical application. PMID- 9752554 TI - [Characterization of liver regeneration in the albumin-urokinase transgenic mouse]. AB - Models of liver regeneration are essential to understand mechanisms of hepatic carcinogenesis, correct genetic diseases by gene transfer or hepatocyte transplantation. The expression in the liver of transgenic mice of a gene coding for a urokinase-type plasminogen activator (uPA mouse) induces hepatotoxicity and prolonged post-native liver regeneration from cellular clones which have inactivated the transgene. This model may have major applications but it remains necessary to characterize the liver regeneration pattern. METHODS: Histological and immunohistochemical studies of the liver of uPA and non-transgenic mice, 3, 7, 14, 21, 28, 42 and 56 days-old. Markers of cellular proliferation: 5-bromo 2'deoxyuridine (BrdU) and proliferating cell nuclear antigen (PCNA). RESULTS: Regenerative nodules were seen from day 14. These nodules then grew, became confluent and by 8 weeks constituted the entire liver mass. A semi-quantitative study of BrdU and PCNA showed a maximal labeling at day 7 (300 to 350 labeled cells/10 microscopic fields, mag 400). When the nodules appeared, 60 to 80% of the cells were labeled. The proportion of labeled cells decreased but was still greater than that observed in non transgenic mice up to day 56 (92 to 106 labeled cells vs 10 to 28, on day 28). CONCLUSIONS: In uPA mouse liver regeneration is significantly expanded, as compared to the regeneration following partial hepatectom. This study therefore has allowed to determine the best conditions for using this model. PMID- 9752555 TI - [Extracorporeal surgery of the renal artery]. AB - PURPOSE: The aim of the study was to assess the short- and long-term results of ex situ renal artery repair in a homogeneous series of patients operated on for complex lesions of this artery. MATERIAL AND METHODS: Seventy-seven patients (38 males and 39 females) underwent 80 extracorporeal repairs of the renal artery. The operated lesions were: aneurysms of the artery and/or of its branches with or without associated dysplasia (30 cases), extensive fibrodysplasia extending to distal branches (31 cases), spontaneous dissection of the artery with extension to the branches (nine cases), reoperation on the renal artery (six cases), miscellaneous (four cases). In all cases, the kidney was exteriorized after transsection of its vessels and cooled by perfusion of cold Collin's solution. After repair, it was reimplanted in the lumbar (36 cases) or iliac fossa (44 cases). An arterial substitute was used in 59 cases. RESULT: No mortality was observed in this series. Five postoperative thromboses occurred leading to kidney loss (6.25%). Segmental thrombosis leading to partial atrophy of the kidney occurred in three cases (3.75%). During the long-term follow-up, one repeat stenosis and four fusiform dilations of venous autografts were observed. All other repairs were successful (89.3%). Results on blood pressure control were favourable in 88.7% of the cases. CONCLUSIONS: Ex situ repair must be reserved to lesions involving several branches of the artery whose repair requires prolonged circulatory arrest and lesions profoundly situated in the renal sinus whose repair is difficult by conventional in situ surgery. PMID- 9752557 TI - [Preoperative portal embolization: an effective means for inducing hypertrophy of the healthy liver and increasing indications for hepatic resection]. AB - AIM OF THE STUDY: The aim of the preoperative portal embolization is a redistribution of the portal venous blood flow in an attempt to induce hypertrophy of the future remnant liver in order to perform a curative liver resection. MATERIAL AND METHODS: Preoperative portal embolization was performed in a group of 43 patients. The volumetric ratio (future remnant liver/total liver tumor) was 20%. Liver metastases were present in 40 patients and primary liver tumor in three. Twenty-four patients had received chemotherapy prior to the preoperative portal embolization. Required operative procedures were right hepatectomy (n = 15), right hepatectomy extended to the segment IV (n = 24) or atypical resection (n = 4). Preoperative portal embolization was performed under percutaneous transhepatic approach with a Blue Histoacryl and Lipiodol Ultra Fluide mixture. Liver volumetric measurements were obtained with 3D color encoded computed tomography, before portal embolization and before surgery. RESULTS: Hypertrophy of the future remnant liver was 83 +/- 58% after a mean 32-day interval between portal embolization and surgery. The tolerance of portal embolization was excellent. Thirty-six hepatectomies were performed as initially planned; seven were cancelled for emergence of metastases (distant in six patients and intrahepatic in one). CONCLUSION: Pre-operative portal embolization is a safe and effective procedure which increases the possibilities of curative resection in the liver tumors. PMID- 9752556 TI - [Possibilities and limits of surgical treatment of malignant adrenal cortex carcinomas. Apropos of a series of 74 cases]. AB - The aim of this study was to report our experience of 74 patients operated on for adrenocortical carcinomas in the last 15 years with particular reference to survival rate and prognostic factors. The tumors were secreting in 78% of the cases and non secreting in 22% of the cases, encompassing stade I: 7%, stade II: 43%, stade III: 21%, stade IV: 29%, according to the MacFarlane classification. All patients were operated on whatever the stade. Adrenalectomy with regional lymphadectomy was performed in 50% of the cases. Other procedures included extended adrenalectomy with nephrectomy (n = 27), hepatectomy (n = 7), and desobstruction or resection of inferior vena cava (n = 13). The resection was curative for 66% of the patients and palliative in 34% of the patients. Local recurrences were operated on in 13 patients. Operative mortality was 4%. Stade I and II had significantly the best actuarial survival rates (78 and 62% at 5 years, respectively), when compared to stade III (27%). Extension to the vena cava was not considered as a contra indication even in cases of massive extension. The survival rate of patients with local recurrences was the same as patients wich stade III: 32% at 5 years. Stade IV tumors had the poorer prognosis with a survival rate of 5% at 3 years. In this group, some patients may have benefited from mitotane. Further studies are mandatory to appreciate the benefit of adjuvant therapy with mitotane and palliative chemotherapy. PMID- 9752558 TI - [Cancer of the thyroid gland with mediastinal extension]. AB - AIM OF THE STUDY: Thyroid carcinomas with upper mediastinal invasion are rare: 24 out of 1,204 thyroid carcinomas operated in the Cancer Center Hospital of Shanghai (2%). For this reason, the report of our experience seems useful. MATERIAL AND METHODS: The 24 patients (13 male and 11 female) had a mean age of 45 years. Ten patients presented symtoms, nine had a palpable cervical mass and in five, the lesion was discovered by an upper mediastinal scan. The operation was performed through an upper partiel sternotomy (n = 12) associated with resection of the internal part of the clavicle (n = 8) and of the first two costal cartilages (n = 4). Tumoral removal was complete in 15 patients and incomplete in nine. RESULTS: There were no peroperative and postoperative deaths. During the procedure, the "brachio-cephalique" vein was injured and sutured in three patients. During the postoperative period, a pneumothorax was observed in three patients and a mediastinal infection in two. When the tumor removal was incomplete (n = 9), postoperative radiotherapy was performed. The only patients with papillary thyroid carcinoma (n = 16) have survived more than 5 years. For the entire group of patients, the 5-year and 10-year survival rate was respectively 65% and 47%. After incomplete tumoral resection followed by radiotherapy, two patients survived with a 28-year follow-up. CONCLUSION: This experience demonstrates that patients may survive beyond 10 years, even after incomplete resection of thyroid carcinoma with upper mediastinal invasion, if the carcinoma is differentiated. PMID- 9752559 TI - [Treatment of gallstones of the common bile duct in the era of celioscopy]. PMID- 9752560 TI - [Patient education by the surgeon. Judicial aspects, ethical aspects, deontologic aspects]. AB - AIM OF THE STUDY: The surgeon must inform his patient about eventual risks before any investigation with diagnosis purpose and before any therapeutic intervention. According to the decree of the first Civil chamber of the "Court of Cassation", the surgeon must be able to prove that this information has been given as required by the article 1315 of the civil code. This decree has created in France an important change in the relationship between surgeon and patient. The French Academy of Surgery has organized a special session to study this problem in terms of legal, ethical and deontological aspects. CONCLUSIONS OF THE ACADEMY: The Academy confirms the necessity to give information to the patients and suggests that the modalities of the information be prepared by the medical societies, the "Conseil National de l'Ordre des Medecins", eventually by ethical committees and legal organizations concerned with medical questions, in view of a common use in all French juridictions. According to the Academy, it is necessary to determine the limits of the information to be given to the patient concerning the most common risks and those with the most serious consequences, in order not to disturb him psychologically. The Academy wishes that necessary and sufficient means be indicated in order for the surgeon to be able to prove that the information has really been given. PMID- 9752561 TI - [Postoperative mortality after resection of pancreatic and periampullary cancers in British specialized units]. PMID- 9752562 TI - [Nissen fundoplication with and without short blood vessels. A controlled prospective study]. PMID- 9752563 TI - [Manual esophagogastric anastomosis or with stapler. Controlled prospective study]. PMID- 9752564 TI - Influencing practices starts with a telephone call. PMID- 9752565 TI - RN first assistants. PMID- 9752566 TI - Laminar airflow systems. PMID- 9752567 TI - Perioperative care of environmentally sensitive patients. AB - In today's complex environment, with an increasing number of chemicals and environmental contamination, some individuals have developed sensitivities to their surroundings. Surgical intervention for environmentally sensitive patients provides an opportunity to reach beyond the boundaries of the OR. These patients require highly individualized perioperative nursing assessments and care planning on a multidisciplinary level. Presbyterian Hospital of Dallas has developed a protocol to initiate collaborative planning for these patients and has had the opportunity to successfully care for these patients. PMID- 9752569 TI - Hospital explores winning balance in perioperative education. AB - Although there are a number of education models used today that expose bachelor of nursing degree students to perioperative nursing, producing nursing graduates who have the education and experience needed to work in the demanding perioperative arena is a challenge for education facilities. The University of Colorado Health Sciences Center, Denver, has developed a perioperative education model that interfaces with one of the university's clinical sites, University Hospital, Denver, to achieve a winning balance between perioperative education and employment. The model provides nursing students with a realistic perspective of the OR and gives the employer an opportunity to thoroughly evaluate potential employees--beyond a resume and references. PMID- 9752568 TI - A criterion-referenced measure of latex allergy knowledge. AB - The exponential use of latex products has increased patients' and health care workers' sensitization to natural rubber latex. The purpose of this research study was to develop and test a set of items designed to measure professional nurses' knowledge about natural rubber latex reactions and interventions appropriate for latex precautions. The items were developed from research reports in the literature and changed to reflect the suggestions offered by experienced perioperative nurses (pilot = 8) and three nationally known experts in latex allergy responses. The revised form was mailed to the home or distributed at the workplace of perioperative nurses (n = 158). Criteria for item retention were < .90 difficulty index and clinical relevance. Certified nurses (i.e., CNORs) scored significantly higher than other nurses who took the test at the same time (t-test = 2.00, P = .05). A pretest and posttest evaluation of the revised 20 items was conducted before and after a latex allergy continuing education program during the spring of 1998. The current knowledge test (i.e., version 1.1) has reliability (alpha = .80); content validity (CVI = .92); and accumulated evidence of construct validity (preinstruction mean = 14.16, postinstruction mean = 17.15, t-paired = 10.54, df = 70, P = < .001). An empirically developed professional nurse latex allergy competency test suitable for clinical use was the outcome of this research study. PMID- 9752570 TI - Moving from an hourly pay model to a professional salary model. PMID- 9752571 TI - Pneumoperitoneum--physiology and nursing interventions. AB - Gaseous insufflation of the abdomen (i.e., pneumoperitoneum) is used routinely during laparoscopic surgery to allow the surgical team members to view the patient's viscera and to perform indicated procedures. Pneumoperitoneum has varying effects on the patient. The initiation and maintenance of pneumoperitoneum causes most of the complications seen with laparoscopic surgery. The perioperative nurse must understand the physiological effects of pneumoperitoneum on the patient to plan appropriate nursing interventions that ensure safe patient care. PMID- 9752572 TI - Involving students in perioperative research. PMID- 9752573 TI - Best practices--ideas that work. PMID- 9752574 TI - Physician-assisted death: can philosophical bioethics aid social policy? PMID- 9752575 TI - Euthanasia and health reform in Canada. PMID- 9752577 TI - Currently accepted practices that are known to lead to death, and PAS: is there an ethically relevant difference? PMID- 9752576 TI - Critical notice: why killing is not always worse--and is sometimes better--than letting die. PMID- 9752578 TI - Terminal sedation as palliative care: revalidating a right to a good death. PMID- 9752579 TI - Assessing the arguments for and against euthanasia and assisted suicide: Part Two. PMID- 9752580 TI - CQ sources/bibliography. PMID- 9752581 TI - Advance directives and voluntary slavery. PMID- 9752582 TI - Gene patents can be ethical. PMID- 9752583 TI - What's so special about the human genome? PMID- 9752584 TI - Disease genes are not patentable: a rebuttal of McGee. PMID- 9752585 TI - Gene patents--a pharmaceutical perspective. PMID- 9752587 TI - On the subject(s) of Jack Kevorkian, M.D.: a retrospective analysis. PMID- 9752586 TI - Disease gene patenting: the clinician's dilemma. PMID- 9752588 TI - Every death is different. PMID- 9752589 TI - Friendly practice guidelines problematic. PMID- 9752590 TI - An ethicist's commentary on the case of the defective puppy adopted out by a breeder. PMID- 9752591 TI - Veterinary research and human health. PMID- 9752592 TI - Salmonella typhimurium DT104: a virulent and drug-resistant pathogen. AB - Salmonella typhimurium phage type (PT) or definitive type (DT) 104 is a virulent pathogen for humans and animals, particularly cattle. It has been isolated increasingly from humans and animals in the United Kingdom and several other European countries and, more recently, in the United States and Canada. Humans may acquire the infection from foods of animal origin contaminated with the infective organism. Farm families are particularly at risk of acquiring the infection by contact with infected animals or by drinking unpasteurized milk. The symptoms in cattle are watery to bloody diarrhea, a drop in milk production, pyrexia, anorexia, dehydration and depression. Infection may result in septicemic salmonellosis and, upon necropsy, a fibrinonecrotic enterocolitis may be observed. The infection occurs more commonly in the calving season than at other times. Feedlot cattle and pigs may also be affected. Prolonged carriage and shedding of the pathogen may occur. Symptoms in humans consist of diarrhea, fever, headache, nausea, abdominal pain, vomiting, and, less frequently, blood in the stool. Salmonella typhimurium DT104 strains are commonly resistant to ampicillin, chloramphenicol, streptomycin, sulfonamides, and tetracycline. PMID- 9752593 TI - [Prevalence of infections caused by Salmonella spp. in cattle and horses at the Veterinary Teaching Hospital of the Faculty of Veterinary Medicine of the University of Montreal]. AB - Bacteriologic detection of Salmonella spp. from feces of animals admitted to Veterinary Teaching Hospital of the Faculty of Veterinary Medicine, University of Montreal, in Saint-Hyacinthe was carried out during a 1-year period to estimate the prevalence of bovine and equine salmonellosis. Prevalence at the time of hospitalization was quite low: 1.4% in cattle and 1.7% in horses. Incidence was 15.1 cases/100 animal/year in cattle and 38.7 cases/100 animal/year in horses. Serotype typhimurium was the most prevalent in both species. In cattle, cases were evenly distributed over the year. In horses, a recrudescence of cases and a obviousness of transmission were apparent in April 1996. PMID- 9752594 TI - A redefined type of elbow dysplasia in the dog--2 cases. AB - Two dogs with calcified masses on the medial aspect of the elbow, resembling those previously reported as ununited medial humeral epicondyle, support proposed ideas that these masses arise due to calcification of flexer tendons at their origin, and that different pathogeneses are likely involved in their development. PMID- 9752595 TI - An unusual manifestation of encephalitozoonosis in chinchilla rabbits. AB - This disease occurred in 6 rabbits in which leukemic changes after infection with the bovine leukemia virus were being studied. PMID- 9752596 TI - Reidentification of Canada's national cattle herd. PMID- 9752597 TI - Chronic selenium toxicosis in growing-finishing pigs in southwestern Quebec. PMID- 9752598 TI - Sensory systems. Web alert. PMID- 9752600 TI - [Clinical evaluation of chemiluminescence immunoassay PSA (ACS-PSA) for detection of prostate cancer]. AB - Serum prostate specific antigen (PSA) levels were measured using an ACS-PSA kit in 147 systematic biopsy cases (61 with prostate cancer (PC)) and 96 transurethral resection of prostate (TUR-P) cases (2 with PC). In the 147 biopsy cases, the sensitivity for PSA using 3.0 and 10.0 ng/ml as cut-off values was 91.8 and 90.2%, while the specificity was 9.30 and 30.2%, respectively. The sensitivity for PSAD (A) (calculated by transabdominal ultrasound) using 0.25 and 0.5 ng/ml/cm3 as cut-off values was 91.8 and 90.2%, while the specificity was 22.1 and 50.0%, respectively. These data indicated that PSAD (A) provided better information for detecting PC than PSA alone. No statistical difference was found between PSAD (A) and PSAD (R) (calculated by transrectal ultrasound) in the utility of detecting PC. PSA below 15.0 ng/ml was seen in sixteen patients with PC. Five of these sixteen patients had a PSA level of < 3.0, and they underwent prostate biopsy based on the abnormality by digital rectal examination (DRE). The other eleven patients had PSAD (A) level of > 0.3 ng/ml/cm3. In all 243 cases, PC was not found in the 49 patients (PSA < 3.0 ng/ml) or 91 patients (PSAD (A) < 0.25 ng/ml/cm3) who had no abnormal findings by DRE and transabdominal ultrasonography. These results suggested a criterion in the use of the ACS-PSA kit for the indication of prostate biopsy and TUR-P. PMID- 9752599 TI - [Long-term results and quality of life of hemi-Kock orthotopic ileal neobladder after radical cystectomy]. AB - We analyzed the long-term results and the quality of life in patients who received orthotopic lower urinary tract reconstruction using the Kock ileal neobladder. Between July 1990 and October 1993, 37 consecutive patients including 2 females received orthotopic hemi-Kock neobladder after radical cystectomy. In these patients, we analyzed the urinary continence, complications and urethral recurrence, and performed a questionnaire survey by mail. Good continence all day had been achieved in 71% of the patients 4 years after surgery. The rate of the pouch-related complications requiring reoperation was 27%. There was no urethral recurrence. Compared with preoperative conditions, 42% were not satisfied with urination. In these dissatisfied patients, the need to use pads in the daytime, sensation of residual urine and weak urine stream were significantly more frequent than in satisfied patients. In summary, the rate of complications was higher than that of other methods. However, the Kock orthotopic ileal neobladder is a stable procedure providing good function over the long-term. PMID- 9752601 TI - [The effects of LH-RH agonist alone or with flutamide in the treatment of stage D2 prostate cancer]. AB - We compared the clinical efficacy of treatment with a luteinizing hormone releasing hormone (LH-RH) agonist alone to combined androgen blockade (CAB) with a LH-RH agonist and fiutamide. A total of 66 stage D2 prostate cancer patients were enrolled from Nov. 1992 to Mar. 1996 (n = 30: LH-RH agonist alone, n = 36 CAB). Serum PSA levels after 3 months of treatment and progression-free survival rates (Kaplan-Meier curves) were compared. Results were statistically evaluated by Wilcoxon's text. There were no differences in PSA levels between LH-RH agonist alone and CAB. Progression-free survival rates were longer in the patients treated CAB compared to LH-RH agonist alone (P = 0.041). Furthermore, in patients with poorly differentiated prostate cancers, longer survival rates were also observed with CAB (P = 0.030). However, there were no differences in high EOD (> or = 2) patients between the two treatments (P = 0.652). PMID- 9752602 TI - [Clinical results of transurethral electrovaporization of prostate (TVP) with two vaporization electrodes]. AB - Transurethral electrovaporization of the prostate (TVP) is a new minimally invasive procedure for treatment of enlargement of prostate. Between April 1996 and September 1997, TVP was carried out in 109 cases with symptoms of lower urinary tract obstruction. A Stortz spike electrode and a Stortz vapor cutting electrode were used for vaporization electrodes. Efficacy parameters evaluated included International Prostate Symptom Score, peak uroflow and postvoid residual volume. By September 1997, 32 cases (spike electrode) and 33 cases (vapor cutting electrode) could be followed up and evaluated. They have shown both subjective and objective voiding improvement. The improvements in subjective symptom scores and objective voiding parameters were not significantly different between the two electrode groups. Early complications included urinary retention, intraoperative burn, stress incontinence, blood transfusion and postoperative hemorrhage. Late complications included urethral stricture, bladder neck contracture and bladder stone. TVP was found to have several advantages, particularly minimal bleeding and the low incidence of postoperative morbidity. The technique is simple and symptoms improve at an early stage following surgery. This study demonstrates that TVP is a safe and effective modality for treatment of BPH. However, long term studies with larger numbers of patients are needed. PMID- 9752603 TI - [Two cases of adrenal myelolipoma diagnosed by ultrasonically guided percutaneous biopsy]. AB - We report 2 cases of adrenal myelolipoma which were diagnosed definitively by ultrasonically guided percutaneous biopsy to waive surgery. In case 1, a 39-year old man presented with abdominal pain. A right adrenal mass was detected by abdominal ultrasonography. In case 2, a 64-year-old man presented with nocturia. A space-occupying lesion above the left kidney was detected by excretory pyelography. Both cases were suspected of adrenal myelolipoma by ultrasonography, computed tomography (CT) and magnetic resonance imaging (MRI). The tumor size was 4 x 4 x 3 cm and 5 x 4.5 x 4 cm in cases land 2, respectively. In both cases percutaneous needle biopsy using ultrasonically guided puncture was performed. Histology showed a complex of hematopoietic and adipose tissues, corresponding to adrenal myelolipoma. The course has been uneventful without surgery for 58 months and 24 months, respectively. Percutaneous adrenal biopsy was useful to establish the definitive diagnosis and to select treatment options. PMID- 9752604 TI - [A case of metastatic renal tumor originating from lung cancer difficult to distinguish from renal cell carcinoma]. AB - A 66-year-old female underwent left lower lobectomy for her lung cancer (adenocarcinoma) in November 1992, followed by the resection of brain metastasis in October 1993. Later, a left renal tumor with paraaortic lymph node swelling was found by a follow-up abdominal CT. She was treated with left nephrectomy and the resection of the paraaortic lymph nodes in June 1997. The histopathology of the resected tumor and the lymph nodes revealed a metastatic renal tumor originating from the pulmonary adenocarcinoma. There have been 38 reported cases of metastatic renal tumor from lung cancer in the Japanese literature. PMID- 9752605 TI - [A case of renal oncocytoma associated with acquired cystic disease of the kidney]. AB - We report a case of renal oncocytoma associated with acquired cystic disease of kidney (ACDK). A right renal mass was incidentally found on an annual ultrasonography of the kidneys in a 56-year-old female patient who had been on maintenance hemodialysis for 6 years. Computerized tomography (CT) showed a right hypervascular renal mass, suggesting a renal cell carcinoma associated with ACDK. Right radical nephrectomy was performed. The post-operative pathological diagnosis was renal oncocytoma. Renal oncocytoma in ACDK is very rare, and the pathological characteristics of oncocytoma are discussed. PMID- 9752606 TI - [A case of renal arteriovenous fistula diagnosed by pharmacoangiography]. AB - A 17-year-old man, who had previously been followed for Nutcracker phenomenon, presented with back pain and gross hematuria. As he developed tamponade of the urinary bladder repeatedly, and the hemoglobin level decreased, we thus performed emergency angiography. A selective left renal venogram revealed the left renal vein to be compressed between the superior mesenteric artery and the abdominal aorta, while the development of an extensive peri- and para-renal collateral circulation was also observed. We next performed epinephrine pharmacoangiography in order to better visualize any abnormal vessels. A left renal arteriovenous fistula was thus diagnosed by this method, and treated successfully by transcatheter embolization using a platinum coil. PMID- 9752608 TI - [Rupture of an infected urachal cyst causing generalized peritonitis: a case report]. AB - A 68-year-old man visited our hospital with complaints of abdominal pain and fever. Physical examination disclosed findings consistent with acute abdomen. Computed tomographic (CT) scan revealed a 5 cm cystic mass contiguous with the dome of the bladder and fluid collection in the peritoneal cavity. Cystogram demonstrated deformity of the bladder and no communication between the mass and the bladder. A diagnosis of generalized peritonitis either due to the infected urachal cyst or ruptured bladder was made, and emergency exploratory laparotomy was carried out. Based upon findings consistent with an infected urachal cyst associated with its intraperitoneal perforation, resection of the entire urachal remnant including the dome of the bladder was performed. Pathologic examination showed an acutely inflammed urachal cyst and chronic inflammation of the bladder wall. PMID- 9752607 TI - [Usefulness of magnetic resonance imaging for evaluation of therapeutic efficacy of bone metastases from bladder cancer: a case report]. AB - A 70-year-old man had a prostate-invading bladder cancer with multiple bone metastases. The bladder cancer seemed to have metastasized via the vertebral vein system because there was no metastasis in other organs. Transurethral resection of bladder tumor was performed followed by one course of M-VAC (methotrexate, vinblastine, doxorubicin) therapy and MEP (methotrexate, etoposide and cisplatin) therapy. After chemotherapy, magnetic resonance imaging (MRI) of the spine revealed further progression of disease, although bone scintigraphy did not, and the patient died of disease. PMID- 9752609 TI - [A case of primary localized amyloidosis of the urinary bladder]. AB - A case of primary amyloidosis of the bladder is reported. A 59-year-old man visited our hospital with a complaint of gross hematuria. Cystoscopy revealed several elevated lesions with yellowish surface accompanied by proliferated vessels at the posterior wall. Transurethral mucosal biopsy was performed. Histopathological diagnosis was a primary localized amyloidosis of AL type in the bladder. Systemic amyloidosis was clinically excluded. He is followed for 38 months without symptoms but mucosal lesions persisted. PMID- 9752610 TI - [A case of colo-vesico-vaginal fistula caused by sigmoid colon diverticulitis]. AB - A 74-year-old woman was referred to our hospital with the chief complaints of pneumaturia, fecaluria and discharge of feces and urine from vagina. Fistulography on the vaginal side showed the presence of contrast medium both in the sigmoid colon and bladder. Colonoscopy revealed multiple diverticulosis of the sigmoid colon. Under diagnosis of colo-vesico-vaginal fistula due to sigmoid colon diverticulitis, a one-stage operation removing sigmoid colon, uterus vaginal wall and urinary bladder wall including the fistula and careful reconstruction was performed. Postoperatively, urinary leakage from vagina in large amounts continued due to the recurrence of vesico-vaginal fistula. An attempt to use human fibrin glue in the recurrent fistula was successful, and the patient was asymptomatic at 21 months of follow-up. Colovesical fistula has been reported in about 10-20% of patients undergoing surgery for complicated diverticulitis, but a combined fistula is a rare condition. Furthermore, we recommend the use of human fibrin glue for a recurrent fistula. PMID- 9752611 TI - [Vesico-vaginal fistula with a giant vesico-vaginal stone: a case report]. AB - The patient, a 73-year-old woman, had undergone hysterectomy and irradiation therapy 26 years ago. On September 4, 1997, the patient was referred to our hospital, and presented with low grade fever and lower abdominal dull pain of a one-month duration. Radiologic and vaginal examinations revealed bilateral hydronephrosis and a giant stone lying down between the bladder and vagina. Vaginal incontinence showed the presence of the vesico-vaginal fistula. She underwent bilateral ureterocutaneostomy and cystolithotomy. A giant vesico vaginal stone was removed by using a hammer and chisel. It weighed 180 g. The stone was composed of calcium phosphate and magnesium ammonium phosphate. Persistent infection of the bladder and the vagina may have been a possible etiological factor of the vesico-vaginal stone formation. Three weeks after the operation, bilateral hydronephrosis was improved. PMID- 9752612 TI - [Giant vesico-vaginal stone: a case report]. AB - A 74-year-old female with the chief complaint of lower abdominal and anal pain had been suffering from total incontinence due to cerebral palsy since her childhood. A giant stone was palpable on vaginal examination. A radiograph showed a giant calcification in the pelvis. Magnetic resonance imaging (MRI) revealed a giant vesico-vaginal stone, which occupied most of the bladder and vagina. Cystolithotomy was performed. The removed stone weighed 435 g, and measured 9.0 x 6.5 x 5.5 cm, and was composed of magnesium ammonium phosphate. To our knowledge only eight cases of female giant vesical stone have been reported. We herein report a rare case of vesico-vaginal stone unrelated to gynecological procedures. PMID- 9752613 TI - [Clinical study on chlormadinone acetate alone followed by combination with LH-RH analogue for prostatic cancer: effects on lipid metabolism]. AB - Twenty-four previously untreated patients with a diagnosis of prostatic cancer were treated with chlormadinone acetate (CMA) alone (100 mg/day) for 4 weeks, and luteinizing hormone-releasing hormone analogue (LH-RHa) was added for the next 24 weeks. Marked decreases in blood LH, testosterone (T), prostate specific antigen (PSA), gamma-seminoprotein (gamma-Sm), and prostatic acid phosphatase (PAP) were observed after a single dose of CMA. T levels were significantly increased 3 days after the initial dose of LH-RHa, and did not return to the pretreatment level. There were no significant increases in any of the markers, nor were there any flare-up cases. Triglyceride levels, which were slightly elevated before the start of treatment, were significantly decreased 24 weeks after the completion of combined therapy. PSA was evaluated as partial response (PR) or better in 86.7% of the patients. Overall evaluation showed PR or better in 75.0% of the patients. These findings suggest that prior administration of CMA followed by combined administration with LH-RHa is useful in the treatment of prostatic cancer. No negative effects on lipid metabolism were observed at any time during the treatment period. PMID- 9752614 TI - Increased plasma epinephrine but not reduced heart rate variability leads to ventricular arrhythmias in patients with acute myocardial infarction. AB - Ventricular arrhythmia observed in the acute stage of myocardial infarction is profoundly related to the autonomic balance. To investigate prediction of ventricular arrhythmia, heart rate variability and plasma catecholamine concentration were simultaneously measured for a week in 17 consecutive patients with first anterior or anteroseptal Q wave infarction treated without specific coronary intervention. The cross-sectional plot of coefficient of variance (= standard deviation of N-N interval/mean N-N interval x 100; %) as a function of plasma epinephrine on the day of admission remained lower than the standard average. Ventricular premature contractions increased in proportion to the plasma epinephrine concentration. In the first week of hospitalization, plasma epinephrine concentration and frequency of premature contraction decreased exponentially, whereas the coefficient of variance showed a modest decline. Ventricular tachycardia refractory to xylocaine with rate accelerating with persistence was observed only in patients with the peak epinephrine concentration > 375 pg/ml. Plasma epinephrine concentration rather than coefficient of variance during sleep after the first acute episode is more closely related to the following triggered ventricular arrhythmia. PMID- 9752615 TI - [Significance of hepatocyte growth factor measurement in patients with acute myocardial infarction]. AB - The use of hepatocyte growth factor for acute myocardial infarction was investigated. Several other biochemical markers are already used for noninvasive detection of acute myocardial infarction. Hepatocyte growth factor, creatine phosphokinase (CK) and CK isozyme (CK-MB) levels were measured in 10 patients with stable effort angina after diagnostic catheterization, and in 21 patients with acute myocardial infarction twice a day for the first 3 days and once a day for the next 4 days. The time to reach the maximum level and the time to decline to less than half of the maximum level for each patient was also measured. Hepatocyte growth factor levels (ng/ml) were 0.3 +/- 0.1 for patients with angina pectoris, and 10.4 +/- 8.8 within 6 hours and 6.7 +/- 4.5 within 12 hours after the onset for patients with acute myocardial infarction. The time to reach the maximum (hours) and the time to decline to less than half of the maximum level (days) were 9.2 +/- 5.2 and 1.1 +/- 0.3 for hepatocyte growth factor, 19.5 +/- 7.2 and 2.3 +/- 1.1 for CK and 16.3 +/- 7.2 and 1.5 +/- 0.4 for CK-MB, respectively. The correlation coefficients between the maximum level of hepatocyte growth factor and the maximum levels of CK and CK-MB were 0.64 and 0.70, respectively. Hepatocyte growth factor is useful as a prognostic indicator and reflects the clinical course in patients with acute myocardial infarction. PMID- 9752616 TI - [Clinical differences between variant and non-variant angina pectoris]. AB - The differences in clinical characteristics were studied between variant angina pectoris with ST segment elevation during ischemic attacks and non-variant angina pectoris without ST segment elevation. Spasm provocation test was performed with either acetylcholine or ergonovine in 192 consecutive patients with vasospastic angina from January 1991 to June 1997. Thirteen patients were excluded because of insufficient data. Fifty-five patients had variant angina and 124 patients had non-variant angina. Coronary risk factors, serum cholesterol level, triglyceride level, high-density lipoprotein cholesterol level, history of syncope, the rates of second or third atrioventricular block and ventricular tachycardia or fibrillation, the incidence of organic stenosis (> or = 50%), the number of vessels with provoked spasm, the dose of acetylcholine and ergonovine used, and duration from the first appearance of chest pain were compared between the 2 groups. Patients with variant angina had more fixed stenosis (p < 0.01), required more percutaneous transluminal coronary angioplasty procedures, lower doses of intracoronary administration of acetylcholine for the induction of coronary arterial spasm and shorter duration from the first appearance of chest pains (p < 0.01) than patients with non-variant angina. However, there were no differences in other factors between the 2 groups. Variant angina pectoris has the same clinical characteristics as non-variant angina pectoris, although variant angina tends to cause higher spasmophilic activity and more fixed stenosis. PMID- 9752617 TI - [Inter-institute variations in International Normalized Ratio and thrombotest]. AB - Oral anticoagulant therapy is effective for reducing the risk of thromboembolic events in patients with atrial fibrillation or other heart diseases. However, the intensity of oral anticoagulation therapy required in high risk patients, especially in Japanese patients, to achieve the best balance between the prevention of thromboembolic events and bleeding complications remains unclear. The multicenter study of Toyama Warfarin Rational Dosage (TOWARD) was started in 1996 to determine the optimal level of anticoagulant therapy. This study investigated the relationship between values of thrombotest (TT) and International Normalized Ratio (INR) measured from the same samples to clarify inter-institute variations. The relationship between TT and INR was not linear but hyperbolic. Changes of INR to TT are relatively small in the TT range of more than 20% as compared with the range of 20% or less. There were considerable inter institute variations of TT, and the coefficient of variation (CV) was 0.16 and 0.24 in the low level and high level anticoagulation samples, respectively. However, the variations became significantly small when the same reference was used. The CV of INR was 0.12 and 0.08 in the high level and low level anticoagulation samples, respectively, and very similar with the control samples without anticoagulation (0.11). The variation was small when INR was obtained from the international sensitivity index (ISI) of thromboplastin less than 1.5. TT is widely used for monitoring oral anticoagulant therapy in Japan, and is an excellent system with little inter-institute variation when a standard reference is offered. Since INR has been established as an international monitoring system, the use of INR measured with thromboplastin of small ISI is recommended for monitoring. PMID- 9752618 TI - [A patient with disopyramide intoxication rescued by percutaneous cardiopulmonary support]. AB - A 28-year-old man was admitted to our hospital in a hypotensive state 2 hours after taking 8,400 mg disopyramide. Infusion of catecholamine and gastric lavage restored normal blood pressure. However, 8 hours after taking the disopyramide he became hypotensive again and electrocardiographic findings revealed bizarre ventricular complexes resulting in ventricular flutter. Although standard cardiopulmonary resuscitation was not effective, his circulatory status was maintained by percutaneous cardiopulmonary support (PCPS). After 36 hours electrocardiography showed sinus rhythm, and his cardiac function became normal. Patients with severe cardiac dysfunction or cardiac arrest caused by disopyramide intoxication can be supported by PCPS until cardiac function is restored. PMID- 9752619 TI - [Coronary angiography provides considerable in vivo pathophysiological information on coronary artery disease]. AB - Coronary cineangiography (CAG) has various limitations and pitfalls, but is widely accepted for evaluating coronary circulation. The interpretation of CAG focuses mainly on the extent and location of coronary artery disease. However, CAG can also provide considerable and varied in vivo pathophysiological information on coronary artery disease as follows: Certain in vivo angiographic coronary morphologies are indicative of histological findings, such as ruptured atheromatous plaque with/without overlying thrombus. Such morphologies are commonly found in patent ischemia- or infarct-related arteries of patients with acute coronary events in both the acute phase and one month after standard medication, and the diseased portion with the morphology is highly specific to the culprit site. The rupture of plaque and overlaid thrombus is just as important in patients with acute coronary events who survive as in patients who die. Diseased sites with complex lesions are prone to progress toward clinical ischemic episodes. The severity of coronary narrowing immediately after acute coronary events does not influence later left ventricular function if antegrade coronary blood flow without distal filling delay is preserved. Coronary stenosis induced by plaque rupture and superimposed thrombus is likely to improve or disappear with time and/or anticoagulant administration with/without antiplatelet therapy. Such information derived from CAG may improve the understanding and suggest optimal therapeutic strategies of coronary artery disease for individual patients. PMID- 9752620 TI - [Cardiovascular imaging in-a-month. Stent implantation for multiple fractures after percutaneous transluminal coronary angioplasty]. PMID- 9752621 TI - Eugenics, contraception, abortion and ethics. PMID- 9752622 TI - Imperialism, research ethics and global health. PMID- 9752623 TI - Bioethics of the refusal of blood by Jehovah's Witnesses: Part 1. Should bioethical deliberation consider dissidents' views? AB - Jehovah's Witnesses' (JWs) refusal of blood transfusions has recently gained support in the medical community because of the growing popularity of "no-blood" treatment. Many physicians, particularly so-called "sympathetic doctors", are establishing a close relationship with this religious organization. On the other hand, it is little known that this blood doctrine is being strongly criticized by reform-minded current and former JWs who have expressed conscientious dissent from the organization. Their arguments reveal religious practices that conflict with many physicians' moral standards. They also suggest that a certain segment of "regular" or orthodox JWs may have different attitudes towards the blood doctrine. The author considers these viewpoints and argues that there are ethical flaws in the blood doctrine, and that the medical community should reconsider its supportive position. The usual physician assumption that JWs are acting autonomously and uniformly in refusing blood is seriously questioned. PMID- 9752624 TI - The cost of refusing treatment and equality of outcome. AB - Patients have a right to refuse medical treatment. But what should happen after a patient has refused recommended treatment? In many cases, patients receive alternative forms of treatment. These forms of care may be less cost-effective. Does respect for autonomy extend to providing these alternatives? How for does justice constrain autonomy? I begin by providing three arguments that such alternatives should not be offered to those who refuse treatment. I argue that the best argument which refusers can appeal to is based on the egalitarian principle of equality of outcome. However, this principle does not ultimately support a right to less cost-effective alternatives. I focus on Jehovah's Witnesses refusing blood and requesting alternative treatments. However, the point applies to many patients who refuse cost-effective medical care. PMID- 9752625 TI - The problematic symmetry between brain birth and brain death. AB - The possible symmetry between the concepts of brain death and brain birth (life) is explored. Since the symmetry argument has tended to overlook the most appropriate definition of brain death, the fundamental concepts of whole brain death and higher brain death are assessed. In this way, a context is provided for a discussion of brain birth. Different writers have placed brain birth at numerous points: 25-40 days, eight weeks, 22-24 weeks, and 32-36 weeks gestation. For others, the concept itself is open to question. Apart from this, it needs to be asked whether a unitary concept is an oversimplification. The merits of defining two stages of brain birth, to parallel the two definitions of brain death, are discussed. An attempt is then made to map these various stages of brain birth and brain death onto a developmental continuum. Although the results hold biological interest, their ethical significance is less evident. Development and degeneration are not interchangeable, and definitions of death apply specifically to those who are dying, not those who are developing. I conclude that while a dual concept of brain death has proved helpful, a dual concept of brain birth still has problems, and the underlying concept of brain birth itself continues to be elusive. PMID- 9752626 TI - The scope for the involvement of patients in their consultations with health professionals: rights, responsibilities and preferences of patients. AB - The degree and nature of patient involvement in consultations with health professionals influences problem and needs recognition and management, and public accountability. This paper suggests a framework for understanding the scope for patient involvement in such consultations. Patients are defined as co-producers of formal health services, whose potential for involvement in consultations depends on their personal rights, responsibilities and preferences. Patients' rights in consultations are poorly defined and, in the National Health Service (NHS), not legally enforceable. The responsibilities of patients are also undefined. I suggest that these are not to deny, of their own volition, the rights of others, which in consultations necessitate mutuality of involvement through information-exchange and shared decision-making. Preferences should be met insofar as they do not militate against responsibilities and rights. PMID- 9752627 TI - Natural deaths while driving: would screening for risk be ethically justified? AB - OBJECTIVES: To determine the epidemiology and the underlying pathological conditions of natural deaths among motor vehicle drivers. Sudden death while driving may cause damage to properties, other vehicles or road users. Although the Medical Commission on Accident Prevention recommended restrictions to drivers at risk of sudden death due to their medical conditions, these restrictions are useless if they do not result in greater safety to the public. DESIGN: A retrospective study of natural deaths of motor vehicle drivers. SETTING: Natural deaths of motor vehicle drivers reported to the coroner for Birmingham and Solihull. SUBJECTS: 86 consecutive natural deaths of motor vehicle drivers in a five-year period between 1984 and 1988. RESULTS: Of the 86 fatalities reviewed, 80 (93%) sudden deaths were caused by ischaemic heart disease. Fifty vehicles were involved in collision with 32 properties, 20 other vehicles and six pedestrians. Fifty-one out of 80 cardiac deaths had past cardiac history and three had reported chest pain prior to the sudden death. CONCLUSION: An applied normative ethical assessment based on the basic moral principles of autonomy, justice, beneficence and non-maleficence are discussed. We conclude that medical screening of drivers has little benefit for the drivers or other persons. PMID- 9752628 TI - The unpaid donation of blood and altruism: a comment on Keown. AB - In line with article 3.4 of EC directive 89/381, Keown has presented an ethical case in support of the policy of voluntary, unpaid donation of blood. Although no doubt is cast on the desirability of the policy, that part of Keown's argument which pertains to the suggested laudability of altruism and of its encouragment by social policy is examined and shown to be dubious. PMID- 9752629 TI - Teaching medical students on the ethical dimensions of human rights: meeting the challenge in South Africa. AB - SETTING: Previous health policies in South Africa neglected the teaching of ethics and human rights to health professionals. In April 1995, a pilot course was run at the University of Cape Town in which the ethical dimensions of human rights issues in South Africa were explored. OBJECTIVES: To compare knowledge and attitudes of participating students with a group of control students. DESIGN: Retrospective cohort study. SUBJECTS: Seventeen fourth-year medical students who participated in the course and 13 control students from the same class, matched for gender. INTERVENTIONS: Students participated in a one-week module on ethics and human rights. Five months after the course had been run, students completed a semi-structured questionnaire exploring their knowledge and attitudes with regards to ethics and human rights issues. MAIN OUTCOME MEASURES: Knowledge scores, attitude scores and various individual indicators of attitude. RESULTS: Clear benefits for overall knowledge score, for four out of five individual knowledge questions and for one of the attitude questions, were demonstrated. Participating students also appeared to be more convinced of the need for teaching on the ethical dimensions of human rights at postgraduate level and that such teaching should also be integrated in the curriculum. The low response rate amongst controls may have selected students who were more socially conscious, thereby leading to an underestimate of the true impact of the course. CONCLUSION: The evaluation indicates clear benefits of the course for undergraduate students, and supports arguments for the inclusion of such courses in the training of health professionals. This is particularly important given the challenges posed by the Truth and Reconciliation Commission to the health professions to address past complicity in human rights abuses through reorientation of medical training in South Africa. PMID- 9752630 TI - Greek theories on eugenics. AB - With the recent developments in the Human Genome Mapping Project and the new technologies that are developing from it there is a renewal of concern about eugenic applications. Francis Galton (b1822, d1911), who developed the subject of eugenics, suggested that the ancient Greeks had contributed very little to social theories of eugenics. In fact the Greeks had a profound interest in methods of supplying their city states with the finest possible progeny. This paper therefore reviews the works of Plato (The Republic and Politics) and Aristotle (The Politics and The Athenian Constitution) which have a direct bearing on eugenic techniques and relates them to methods used in the present century. PMID- 9752631 TI - Tensions in setting health care priorities for South Africa's children. AB - The new South African constitution commits the government to guarantee "basic health services" for every child under 18. Primary health care for pregnant women and children under six and elements of essential primary health care have received priority. At present, there is little analysis of the moral considerations involved in making choices about more advanced or costly health care which may, arguably, also be "basic". This paper illustrates some of the tensions in setting priorities for a just macro-allocation of children's health care, given the realities of need and scarce resources, and the commitment to equality of basic opportunities. PMID- 9752632 TI - Ethics and aims in psychotherapy: a contribution from Kant. AB - Psychotherapy is an activity which takes many forms and which has many aims. The present paper argues that it can be viewed as a form of moral suasion. Kant's concepts of free will and ethics are described and these are then applied to the processes and outcome of psychotherapy. It is argued that his ideas, by linking rationality, free will and ethics into a single philosophical system, offer a valuable theoretical framework for thinking about aims and ethical issues in psychotherapy. PMID- 9752633 TI - "Goodbye Dolly?" The ethics of human cloning. PMID- 9752634 TI - Decision making in the critically ill neonate. PMID- 9752635 TI - Causing death or allowing to die? A rejoinder to Randall's comments. PMID- 9752636 TI - The ethics of human cloning. PMID- 9752637 TI - A randomized prospective study comparing supportive and dynamic therapies. Outcome and alliance. AB - The authors report preliminary results of Brief Supportive Psychotherapy (BSP) in the Beth Israel Brief Psychotherapy Program for a sample with primarily Cluster C Axis II disorders. This study compares 24 patients treated with BSP with 25 patients treated with Short-Term Dynamic Psychotherapy (STDP). STDP was chosen because its confrontational methods contrast dramatically to BSP, which emphasizes building self-esteem, reducing anxiety, and enhancing coping mechanisms. Videotaped therapies were based on manualized 40-session protocols. Similar degrees of improvement were seen in BSP and STDP at termination and at 6 month follow-up. A study of therapeutic alliance in BSP showed stable and high levels of alliance in good-outcome cases and more variability in poor-outcome cases. These preliminary findings are consistent with other studies and suggest supportive psychotherapy may be effective for many patients, leading to significant and lasting change. PMID- 9752638 TI - A time-limited behavioral group for treatment of obsessive-compulsive disorder. AB - In vivo exposure with response prevention is an effective treatment for obsessive compulsive disorder (OCD) either alone or combined with pharmacotherapy. Widespread application of this technique has been limited by lack of trained therapists and the expense of intensive individual behavioral therapy. This report describes a time-limited 10-session behavioral therapy group for OCD whose key elements are exposure, response prevention, therapist and participant modeling, and cognitive restructuring. In a naturalistic open trial of 90 patients meeting DSM-III-R criteria for OCD who completed the 10-session group, self-administered Yale-Brown Obsessive-Compulsive Scale scores (mean +/- SD) were 21.8 +/- 5.6 at baseline and 16.6 +/- 6.4 after the 10-week treatment, a significant decrease. A descriptive analysis of the therapeutic elements of the group and its advantages over individual behavioral treatment are presented. PMID- 9752639 TI - Religion and the psychotherapeutic relationship. Transferential and countertransferential dimensions. AB - The salience of religion in society and health care has received increased attention. Recent developments in psychiatry reflect a broader view of religion that includes an appreciation of its adaptive and maladaptive dimensions. An examination of religious countertransferential and transferential reactions provides a framework for examining religious themes. Case examples illustrate the following critical factors that increase therapists' skill in working with religious themes: 1) monitoring the therapist's own attitude toward religious content, 2) attending to religious content, 3) seeking consultation, and 4) using religious content in interpretations. PMID- 9752640 TI - Therapist interventions in early sessions of brief supportive-expressive psychotherapy for depression. AB - Although psychotherapy manuals provide treatment guidelines, detailed descriptions of therapist interventions in manual-guided therapies are lacking. The purpose of the present investigation was to evaluate the types of therapist interventions in Supportive-Expressive (SE) psychotherapy for depression by using a molecular method of assessment and then to compare the results with those attained with a molar method. Four percent of therapist statements per session early in treatment were interpretations, which most often focused on the patient's parents, significant others, and self in the present time frame. This molecular method for assessing therapist interventions did converge with the molar adherence/competence method. PMID- 9752641 TI - Differential effects of interventions on the therapeutic alliance with patients with personality disorders. AB - The goal of this study was to examine the relationship between clearly defined therapist interventions and the therapeutic alliance with personality-disordered patients. Transcripts of one psychotherapy session for each of 5 subjects taking part in a long-term psychotherapy research project were rated for therapist interventions and therapeutic alliance to determine if specific interventions were followed by enhanced or diminished therapeutic work. Transference interpretations were followed by a deterioration in the therapeutic alliance when the alliance was weak, but by enhanced work when the alliance was solid. In patients with both strong and weak alliances, defense interpretations and supportive interventions enhanced therapeutic work without increasing defensiveness. Supportive interventions seemed to prepare the way for exploration and to repair ruptured alliances. PMID- 9752642 TI - The medial collateral ligament of the elbow joint: anatomy and kinematics. AB - Eighteen osteoligamentous elbow joint specimens were included in a study of the medial collateral ligament complex (MCL). The morphologic characteristics of the MCL were examined, and three-dimensional kinematic measurements were taken after selective ligament dissections were performed. On morphologic evaluation the MCL is divided into the anterior bundle and the posterior bundle. The anterior bundle can be divided into anterior and posterior bands. The maximum valgus and internal rotatory instability after transection of the anterior band, 11.7 degrees and 11.2 degrees, respectively, were found at elbow flexions of 30 degrees and 40 degrees. Severance of the entire anterior bundle produced major valgus and internal rotatory laxity through the complete flexion arc of maximal 14.2 degrees and 18.5 degrees, respectively, at 70 degrees and 60 degrees of elbow flexion. Cutting both the posterior band and the posterior bundle resulted in only internal rotatory laxity of maximal 7.2 degrees at 130 degrees of elbow flexion. This study defines the anterior band as the primary constraint to valgus and internal rotatory forces, the posterior band as the secondary, and the posterior bundle as the tertiary constraint. The MCL was observed to be a complex of ligamentous fibers rather than individual bands that stabilizes the joint against valgus and internal rotatory forces. PMID- 9752643 TI - The pattern of pain produced by irritation of the acromioclavicular joint and the subacromial space. AB - To characterize the patterns of pain caused by selective irritation of the acromioclavicular joint and of the subacromial space, hypertonic saline solution was injected 15 times into the acromioclavicular joints of 10 healthy volunteers and 10 times into the subacromial space of 9 healthy volunteers. Irritation of the acromioclavicular joint produced pain directly over the joint, in the antero lateral neck, in the trapezius-supraspinatus region, and in the anterolateral deltoid. Irritation of the subacromial space produced pain in the region of the lateral acromion, the deltoid muscle, and occasionally in the forearm or the fingers but did not produce pain in the neck or in the trapezius region. Neither acromioclavicular nor subacromial irritation produced pain at the posterior aspect of the shoulder. This information may assist in accurate clinical diagnosis and in the selection of optimal imaging studies for the evaluation of shoulder pain. PMID- 9752644 TI - Glenoid bone architecture. AB - This article describes regional variations in trabecular bone architecture in terms of density and orientation within six glenoid specimens. The mean donor age was 56 years and ranged from 31 to 72 years. An automated imaging technique based on 3-dimensional serial sectioning was used for the direct examination of the glenoid cancellous bone structures. Subchondral plate thickness was on average 1.9 mm and ranged from 1.2 mm to 2.9 mm. The volume fraction of trabecular bone varied from 11% to 45% with peak values at the posterior glenoid vault. On graphic 3-dimensional reconstructions, the glenoid appeared as platelike trabeculae, radially oriented perpendicular to the subchondral plate and interconnected by thin rods. These views also displayed regional variations throughout the glenoid, reflecting differences in the macroscopic appearance. Quantitative structural analysis revealed different degrees of anisotropy at the glenoid cancellous region, predominantly transverse isotropy. Resemblance to direct weight-bearing cancellous bone such as the proximal tibia was evident. PMID- 9752645 TI - Shoulder arthroplasty in recreational golfers. AB - A retrospective review of 24 patients who had shoulder arthroplasty revealed that 23 were able to resume playing golf. The 23 patients (mean age 52.4 years, range 26.4 to 71.9 years) underwent 26 shoulder arthroplasties, 20 total shoulder arthroplasties, and 6 hemiarthroplasties. The average follow-up was 53.4 months (range 24.4 to 127.2 months). The average length of time from shoulder arthroplasty to playing an entire round of golf was 4.5 months. Eighteen patients were able to report their preoperative handicap and noted an average improvement after surgery of almost 5 strokes. Playing golf did not result in increased radiographic evidence of component loosening, and no increase occurred in lucent lines when the golfers were compared with a control group of 76 patients with osteoarthritis who had 103 shoulder arthroplasties (P < .05). Fifty members of the American Shoulder and Elbow Society were mailed a standardized questionnaire of 11 questions concerning patients who had shoulder arthroplasty and played golf. Most surgeons (91%) encouraged such patients to resume playing golf. The average length of time members of the Society recommended that patients should wait after shoulder arthroplasty before resuming golf was 4.3 months. Approximately 60% of surgeons believed that no limit should be placed on the number of golf rounds played weekly, and 91% denied an increase in complications among those who returned to playing golf after undergoing shoulder arthroplasty. Fewer than one third of the surgeons (29.5%) believed that component wear would be a problem in patients who undergo shoulder arthroplasty and play golf frequently and would recommend a hemiarthroplasty for an active golfer because of concern about future glenoid problems. PMID- 9752646 TI - Morphologic changes in the ulnar nerve at the elbow with flexion and extension: a magnetic resonance imaging study with 3-dimensional reconstruction. AB - We evaluated the morphology of the ulnar nerve and cubital tunnel with noninvasive magnetic resonance imaging (MRI). We used fresh human cadavers with the elbow in full extension, 90 degrees of flexion, and full flexion. For each elbow, 1-mm slices were imaged interpolated, and reconstructed into 3-dimensional data volumes, and then manually segmented before they were examined with sequential transverse sections, curved sections, and 3-dimensional images. The ulnar nerve follows a tortuous course in full extension, becomes progressively linear with incremental elbow flexion, shifts anteriorly in the cubital tunnel, and flattens against the medial epicondyle. The proximal and midportions of the cubital tunnel also change with flexion from round to elliptical. In addition, successive increases occur in the cross-sectional diameter of the mediolateral plane. The nerve is surrounded by fat throughout the cubital tunnel except adjacent to the medial epicondyle. These observations suggest that the ulnar nerve progressively stretch over the medial epicondyle occurs when the normal elbow is flexed. Direct compression areas of the ulnar nerve were not seen in our study of normal human elbows. PMID- 9752647 TI - Structural changes of the rotator cuff caused by experimental subacromial impingement in the rat. AB - Subacromial impingement of the infraspinatus tendon was experimentally created in 28 young adult rats by thickening the undersurface of the acromion with either one or two platelike bony transplants of the ipsilateral scapular spine. Nine nonoperated and eight shoulders that had undergone a sham operation served as control groups. The rats were killed after 2 days and after 1, 2, 4, 8, 16, and 32 weeks for histologic evaluation. All rats with experimental subacromial impingement showed an infraspinatus tear on the bursal side of the tendon. An isolated tear on the articular side or within the tendon was not seen. Two plates caused larger tears than one (P = .04), and more long-standing impingement was associated with larger lesions (P = .002). Multiple chondrocytes were observed within the tendon adjacent to the bony transplants. No calcium deposits were found. In the subacromial space rapid thickening of the bursa was observed. The undersurfaces of the bony transplants showed no evidence of abrasion or remodeling caused by the tendon. The shoulders of the control groups were found intact without any alteration. Experimental subacromial impingement in the rat caused bursal side rotator cuff tears. The type of partial tears that are most frequently observed in clinical practice, that is, intratendinous and articular side tears, were not seen in this experimental model. PMID- 9752648 TI - Articular contact patterns of the normal glenohumeral joint. AB - The purpose of this study was to determine the articular contact patterns of the normal glenohumeral joint, and to correlate these findings with cartilage and subchondral bone architecture. We studied 10 normal shoulders of cadavers. We removed all soft tissues except the joint capsule and rotator cuff and then placed the shoulders on a testing apparatus that allowed freedom of translation in three planes. After the humerus was placed in a neutral position of rotation, articular contact patterns were measured with specially prepared prescale Fuji film so that it could be inserted between the joint surfaces. Articular contact was analyzed with 222 and 444 N of joint compressive load, and the humerus was positioned in scapular plane abduction of 0 degree, 45 degrees, and 90 degrees. The contact patterns were then digitized to determine percentage contact of the humeral head on the glenoid. We studied 12 additional cadaver shoulders with fine microradiographs and histologic techniques after we sectioned the glenoids in the anterior-posterior and superior-inferior planes. We then analyzed articular and subchondral architecture. We found that when the shoulder was adducted the contact area of the humeral head on the glenoid was limited to the anatomic region of the central glenoid known as the "bare area." This was histologically and radiographically an area of cartilage thinning and increased subchondral bone density. As the shoulder was abducted the articular congruity and percentage contact area increased. We concluded that there was a slight articular mismatch with the shoulder adducted in the normal shoulder. Histologic and radiographic studies suggested that the central bare area region of the glenoid was a region of increased compressive loading. As the shoulder was abducted the joint became more congruent and thus the contact area of the humeral head on the glenoid increased. PMID- 9752649 TI - Spontaneous detachment of the deltoid muscle origin. AB - Three patients with four affected shoulders with spontaneous detachment of the deltoid origin are reported. In this group spontaneous detachment of the deltoid origin was associated with chronic massive rotator cuff defects. Detachment of the deltoid origin was associated with an acute, sudden onset of shoulder weakness. Pain was not a prominent complaint, and shoulder function was poor because of weakness. Spontaneous detachment of the deltoid origin can occur with chronic massive rotator cuff tears and can be responsible for an acute onset of shoulder weakness. PMID- 9752650 TI - Passive tension in the supraspinatus musculotendinous unit after long-standing rupture of its tendon: a preliminary report. AB - Incomplete functional recovery after rotator cuff surgery can be caused by rerupture or incomplete restoration of the contractile properties of the muscle tendon-bone unit. We measured the passive tension generated in the supraspinatus musculotendinous unit at the time of repair of the supraspinatus tendon performed for the treatment of long-standing rupture in four patients and compared our results with the values of an intact musculotendinous unit. In stepwise elongation from 10 to 20 mm, passive tension increased by a factor of 2.2 +/- 0.4 in the study group. In the control case passive tension increased by a factor of only 1.3. Mean tension in 60 degrees of abduction was 14.25 +/- 3.4 N in the four long-standing ruptures and 10 N in the control case. If the arm was brought to the side, tension rose to 25 N in the control case, whereas mean tension increased to 59.25 +/- 12.7 N in long-standing rupture of the supraspinatus muscle. Our findings demonstrate that passive tension in the supraspinatus is increased after long-standing rupture of its tendon. This result suggests that active force generation by this muscle will be compromised after surgery and that the high strain after repair may expose the musculotendinous unit to further damage. PMID- 9752651 TI - The mechanism of creation of superior labrum, anterior, and posterior lesions in a dynamic biomechanical model of the shoulder: the role of inferior subluxation. AB - Lesions of the superior glenoid labrum are a source of shoulder disease. However, the mechanisms of injury to this region are unknown, and controversy exists regarding the role of shoulder instability in creation of this lesion. With a cadaver model that simulates physiologic rotator cuff forces and produces traction on the biceps tendon, the creation of type II superior labrum, anterior, and posterior (SLAP) lesions and the role of glenohumeral subluxation were investigated: Left and right shoulders from each of 8 paired cadavers (age 62 +/- 7.2 years, 5 male and 3 female) were randomized to be tested in either a 20 mm inferiorly subluxed position or in a reduced position. The long head of the biceps tendon was held near the musculotendinous junction with a novel cryogenic clamp. Traction on the long head of the biceps tendon was applied at a fast rate of 12.7 cm/sec with a servohydraulic testing machine. A load cell was used to monitor the biceps tendon load. After testing to failure, the presence or absence of a type II SLAP lesion was determined by 2 experienced shoulder surgeons masked to the test group. The production of type II SLAP lesions differed significantly (P = .03) between reduced shoulders (2 SLAP lesions out of 8 tests) and the shoulders with inferior subluxation (7 SLAP lesions out of 8 tests). This experiment has shown that traction on the biceps tendon in this biomechanical model can reproducibly create type II SLAP lesions, and inferior subluxation facilitates the generation of such lesions. PMID- 9752652 TI - Extension osteotomy in malunited clavicular fractures. AB - The association of a malunited clavicular fracture with a pattern of disability that includes not only pain but also impairment of shoulder function is rare. But in cases where such an association exists, correction of the clavicular deformity should be considered. We report on 4 patients with a malunited fracture of the clavicle after nonoperative treatment. In all 4 patients fracture union had occurred with shortening associated with pain and ipsilateral glenohumeral dysfunction. The deformity was not associated with neurovascular dysfunction. On preoperative radiographs the shortening of the malunited clavicle was between 0.9 and 2.2 cm compared with the contralateral clavicle. all patients underwent an extension osteotomy of the clavicle with interposition of an autogenous iliac crestbone graft secured with a plate and screws. The length of follow-up was 1 to 4 years in 3 cases and 6 weeks in 1 case. The functional outcome was evaluated with the Constant-Murley and University of California-Los Angeles scales. All patients had immediate pain relief after osteotomy and correction of the deformity. Shoulder function rapidly improved and functional outcome was good in 3 of the 4 patients. In cases of shoulder function impairment associated with malunited clavicular fractures, extension osteotomy combined with autogenous bone grafting is likely to produce good results. PMID- 9752653 TI - Periprosthetic humeral fractures: mechanisms of fracture and treatment options. AB - In 20 patients, 21 periprosthetic humeral fractures were reviewed retrospectively. The mean follow-up time was 27.1 months. Mild osteopenia was present in 45% of the patients, whereas 30% had severe osteopenia. Five mechanisms of fracture were identified, including 3 intraoperative causes that are avoidable. Treatment with stable intramedullary fixation utilizing the humeral stem and cerclage wiring provided superior results in terms of time to union, adverse effect on rehabilitation, and occurrence and severity of surgical complications. Diaphyseal fractures that were treated with standard stem arthroplasty with or without supplemental fixation had a longer time to fracture union, a higher complication rate, and prolonged rehabilitation. Fractures of the proximal humeral metaphysis can be treated with standard stem arthroplasty and cerclage wiring if the stem extends distal to the fracture site by at least 3 cortical diameters. Anatomic reduction of fractures treated by surgical means results in shorter healing times. Cast or brace immobilization can be used for management of postoperative fractures that occur distal to a well-fixed and stable prosthetic stem. Cast or brace immobilization results in fracture union but rehabilitation may be greatly impaired, and there is an increased risk of complications associated with immobilization of the extremity. Long-stem intramedullary fixation with cerclage wiring is the preferred surgical option for treatment of unstable humeral shaft fractures. PMID- 9752654 TI - Fixation of humeral prostheses and axial micromotion. AB - Surgeons often avoid cementing a proximal humeral prosthesis. Occasionally bony augmentation is needed. This study was undertaken to compare proximal cementation in combination with distal press with total cementation or press fit alone. In phase 1 axial micromotion with axial loading was measured in 15 pairs of humeri: 5 fully cemented versus proximally cemented, 5 fully cemented versus press fit, and 5 proximally cemented versus press fit. X-ray films of the specimens were obtained to assess canal fill. In phase 2 axial micromotion was measured in 5 pairs of high mineral density and 5 pairs of low mineral density to compare proximal cementation with press fit. The 3 M modular prosthesis was used in both phases. No difference was found in phase 1 among the 3 fixation techniques. A strong reverse statistical correlation (P = .007) (r = .55) was seen between axial micromotion and fill of the canal with the prosthesis. In the second phase no statistically significant difference was found between the techniques of fixation or between the 2 bone densities. Fill of the canal at the distal end of the prosthesis was the only variable found that affected axial micromotion, but it accounted for only approximately 30% of the variance. Bone quality and augmentation of the proximal bone with cement did not affect axial micromotion in this experiment but warrant further study. PMID- 9752655 TI - The dynamic elbow suspension splint. AB - Elevation of the upper extremity after elbow surgery has rarely been advocated and can be difficult to achieve. Usually the extremity is elevated with the elbow at 90 degrees of flexion, so that swelling from the hand drains to the elbow but the elbow remains dependent. Excessive swelling causes discomfort and compromised wound healing, makes early mobilization difficult, and predisposes to joint contracture. We report on a dynamic elbow suspension splint, which is analogous to the Thomas splint used for femoral shaft fractures. The arm is held in full extension with an above-elbow plaster slab and is secured to the Thomas splint with skin traction. The splint is suspended on a Balkan frame at an angle of 60 degrees. We prefer to use the new Zimmer Thomas splint, because it is radiolucent and has self-adhesive sheep-skin supports that can be simply applied. It allows the patient to mobilize in bed and is well tolerated by patients and nursing staff. The dynamic elbow suspension splint is a useful adjunct after complex elbow surgery or trauma, because it reduces swelling and maintains the elbow in extension. PMID- 9752656 TI - Aseptic loosening of the humeral component in total shoulder arthroplasty. PMID- 9752657 TI - Snapping elbow caused by the synovial fold in the radiohumeral joint. PMID- 9752658 TI - Chondrosarcoma of the scapula in an adolescent girl. PMID- 9752659 TI - Supraspinatus rupture at the musculotendinous junction. PMID- 9752660 TI - Acromioclavicular joint cyst and rotator cuff tear. PMID- 9752661 TI - Surgical removal of pseudotumoral calcified bursitis of the shoulder in a patient with hemodialysis. PMID- 9752662 TI - Thoracic outlet syndrome. AB - A careful history and thorough physical examination are the most important components in establishing the diagnosis of TOS. The use of radiographic and laboratory tests, when indicated, can improve the diagnostic yield. Provocative positional maneuvers must be evaluated for their vascular and, more important, for their neurologic response. These maneuvers do not "make" the diagnosis, but they can be a useful adjunct for confirming the diagnosis. Vascular TOS is less common and often requires surgical treatment. Neurologic TOS is very common but less frequently requires surgical treatment. The surgical approach and procedure selection should be determined by the nature of the pathologic condition and site of compression. PMID- 9752663 TI - [Data from an expedition to study a Siberian vegan settlement]. AB - Health status, the way of life and nourishment of 84 vegans in Siberian village (Krasnoyarsk region) were studied and compared with those of 26 meat-eaters. The investigation included work with a questionnaire, clinico-diagnostic and laboratory research. It was shown that a vegetarian diet improves the serum lipid spectrum (cholesterol, LPLD, cholesterol of LPNP, atherogenic coefficient), normalizes weight and cardiovascular system. The vegans had normal levels of vitamin B12 and serum Fe but the calcium level in this group was lowered as compared with the control group. The pathology of internals (nephroptosis, lithic diathesis, tendency to lithogenesis) was observed. Apparently, the high serum Zn levels found in both groups aren't directly caused by the diet but by climate and geographic factors. PMID- 9752664 TI - [Development of a method of studying actual nutrition according to analysis of the frequency of consumption of food products: creation of a questionnaire and general evaluation of the reliability of the method]. AB - The method of actual feeding evaluation on frequency of food-stuffs consumption was developed. The method of 24-hour reproduction of a feed and the method of consumed food registration in a diary by testing person himself. The main results of study testified to reliability of data, received by a developed method. PMID- 9752665 TI - [A new method of evaluating the utilization of nutrients (carbohydrates, amino acids and fatty acids) on the plastic and energy goals in the animal body]. AB - With use of a new method, based on detection in blood serum of radioactivity of water, formed from tritium marked precursors--glucose, amino acids (valine, serine, histidine) and palmitine acid--their distribution on oxidizing and anabolic ways of metabolism was determined. The work was carried out on laboratory rats. In young pubertal rats the ratio of flows on these ways for glucose was found equal 2.83, i.e. it in a greater degree was used as energy substratum. On the contrary, for palmitine acid this ratio was equal 0.10--it was comprised in a plastic material of organism in a greater degree. For serine, histidine and valine it is equal 0.34, 0.71 and 0.46, accordingly. In growing rats the distribution of flows was shifted aside of anabolic way: the ratio of flows is equal 0.19; in old rats--aside of oxidizing: a ratio of flows is equal 0.71. PMID- 9752666 TI - [Effect of a bioactive food supplement from selenium enriched baker's yeast autolysate on the status of the intestinal barrier in rats with anaphylaxis]. AB - Rat's intestinal barrier permeability disturbed in consequence of intestinal anaphylaxis reaction was almost completely normalized in animals fed with baker's yeast autolysate "Vitasil" enriched with selenium on a level of 3 mg Se/day during 29 days. These rats showed in comparison to Se-unsupplemented animals a significant elevation of Se level in red blood cells and plasma together with a decrease of intestinal mucosal TCA-soluble thiol compounds. Urinary Se excretion was significantly elevated in comparison to unsensitized rats both in Se supplemented and unsupplemented animals with anaphylaxis. It's concluded that "Se Vitasil" may be successfully used in antioxidative therapy of food allergy, malabsorption, inflammatory bowel diseases and intestinal infection. PMID- 9752667 TI - [The prevalence of iron-deficiency anemia in young women in the Republic of Tajikistan]. AB - The nutrition status and some obstetric-gynaecological factors influencing on anemia prevalence among young women in Tadjikistan was estimated. It was shown, that the daily average consumption of energy by the young women is lower than FAO/WHO and Institute of Nutrition of RAMS recommended. 36.9% of women consumed protein below of WHO established safe level. Many persons of examined group had iron-deficient status. PMID- 9752668 TI - [Functional properties of alginates and their use in therapeutic-prophylactic nutrition]. AB - Adsorbability of alginic acid and its salts extracted from brown seaweeds and character of their biological action were studied. The results of clinical tests alginic acid and its salts as well as of preparations and food-stuffs, prepared from Laminaria japonica Arech, were reflected. The potentialities of use of alginates and food-stuffs containing them in therapeutic nutrition was shown. PMID- 9752669 TI - [Dynamics of postprandial glycemia in patients with insulin-dependent diabetes mellitus as affected by extruded and micronized products from cereal and big crops]. AB - The dynamic of postprandial glycemia in patients with insulin-dependent diabetes under influence of extruded and micronized food-stuffs prepared from cereals was investigated. It was shown extruded and micronized food-stuffs render various influence on post-eat glycemia dynamics and significantly differed in glycemic index value. PMID- 9752670 TI - [Biologically active additive from milk thistle in the solution of public health problems]. AB - Influence of bioactive additions from natural vegetable raw materials--silybum marianum--in the content of bread products on health and metabolism was studied. It is shown bread products with silybum marianum to be effective medicine with general restorative influence, increasing internal protection resources, capacity for work and vital activity. PMID- 9752671 TI - [Dietary fiber-- a new chapter in the chemistry and technology of food]. AB - In the last years the number of publications, devoted to composition, structure, properties, technology and use of food fibres, has much increased. It is caused their important physiologic role in a feeding and functions of digestion system and extending potentialities of them introduction as the additives in various kinds of food-stuffs. It is possible already today to allocate all accumulated information on food fibres in independent section of chemistry and technology of food and to carry out their systematization. Proposed systematization allows to expand opportunities of creation and allocation of the composition additives on the basis of food fibres, to establish interrelation between a structure, properties, medical-biological efficiency of these additives, to solve problems of formulas and sales of new kinds of food and etc. PMID- 9752672 TI - [Dependence of nutritive value of canned meat on the thermal of the raw materials and electric stimulation]. AB - Research of influence electric stimulation and different thermal condition of raw material on meat protein breaking down by proteinases of gastrointestinal tract (pepsin, trypsin and chymotrypsin) was carried out. It was shown that electric stimulation increased the rate of meat protein proteolysis significantly, i.e. from the point of view of breaking down of meat protein by enzymes it gave effect, similar meat ripening in the cool condition. PMID- 9752673 TI - [Electrophoretic methods in the analysis of food products]. PMID- 9752674 TI - [Cholesterol and atherosclerosis]. PMID- 9752675 TI - [Benzodiazepines: history and contemporary issues]. PMID- 9752676 TI - [The diagnosis, treatment and prevention of early stages of cerebral blood flow insufficiency]. AB - Biomicroscopy of bulbar conjuctiva as well as oencephalography were quite informative for discovery of subclinical manifestations of disorders of cerebral circulation (DCC). That conclusion resulted from the observation of 133 patients with early forms of vascular pathology of brain. Besides, to find the early signs, it was also worth while to perform some biochemical studies (coagulogram, studies of both rheologic properties and lipids of blood). In order to establish discirculatory encephalopathy it was also expedient to use ultrasonic dopplerography and electroencephalography. Efficiency of some medical prophylactic measures (normalization of the diet, weight, muscular activity, administration of antiatherosclerotic drugs) that prevented the progression of aorta's stenosis was also demonstrated. PMID- 9752677 TI - [The syndrome of depressive depersonalization]. AB - Nineteen patients (9 men, 10 women aged 22--38 years) with depressions were examined. The phenomena of anesthesia of ideatoric functions dominated in clinical picture of such depression. This depression is defined as the depression of estrangement. Psychopathologic differentiation of mental estrangement in the structure of depression and in similar disorders of self-consciousness which were formed in depressions under conditions of "transitional syndrome" (according to G.Gross) is outlined. This syndrome is characterised by irreversible negative disorders of "defective depersonalization". The following signs permit to distinguish depression of estrangement from "transitional syndrome": partiality of the estrangement's phenomenon; connection between psychopathologic formations and pathology of imagination (figurative expressiveness, demonstrativity, lability to psychogenic and medical actions); conformity of the syndrome's structure and characteristics of premanifested personality's structure (combination of hyperthymic features with histrionic and/or narcissic ones). "Apperceptive anesthesia" is suggested for designation of the variant of the depression described. PMID- 9752678 TI - [The treatment of patients with hypertension-related intracerebral hematomas]. AB - The paper presents the results of both conservative and surgical treatment of 300 patients with hypertension, complicated by intracranial hematoma. It is shown that the results of therapy depend on the size and location of hematoma, on the severity of both perifocal disorders and dysfunction of truncal structures as well as on the dynamics of arterial pressure and degree of somatic burden. PMID- 9752679 TI - [The role of personality aspects in patients with chronic neurological symptoms of vertebral osteochondrosis and psychological rehabilitation methods]. AB - The aim of the study is to define mechanisms of influence of the most typical personal traits on chronization of vertebroneurological signs. 100 chronic patients with vertebroneurologic pathology were examined. Early signs of psychic dysadaptation, hypostenic or mixed types of psychological reactivity, low lability of intellectual processes and nonspecific signs were found in the most of the patients. It is suggested that insufficiency of psychological defence might contribute to the development of psychic dysadaptation. Both mechanisms of psychological defence and attitude toward disease were conditioned by predominant personal peculiarities. It is personal characteristics that cause predisposition to development of psychic dysadaptation and to unfavourable attitude toward disease. That in turn contribute to the development of the disorders of motor stereotype. Psychocorrection of such patients is discussed. PMID- 9752680 TI - [The changes of lipids, lipoproteins and apolipoproteins in plasma in ischemic stroke]. AB - Contents of lipids (cholesterol CS, triglycerides) as well as levels of CS of lipoproteins of different density and of apolipoproteins A1 and B were measured in blood serum of 31 patients in different periods after of ischemic stroke. The majority of the indices studied were significantly decreased in men in acute periods of the stroke, especially in 8-21 days after the stroke development. Meanwhile, the levels of lipids, CS lipoproteins of both low and very low densities were increased later. Contents of CS of antiatherogenic lipoproteins of high density (HDLP) was low in different periods after the development of disorders of cerebral circulation. This confirms the concept that low level of HDLP is one of the risk factors of ischemic stroke. Decrease of the levels of both lipids and CS of lipoproteins of different density was more pronounced in patients with more severe atherosclerotic damages of the main cerebral arteries. Disorders of metabolism of lipoproteins in ischemic disorders of cerebral circulation are discussed with reference to literary data taking into consideration their heterogeneity. Genetically determined pathology of certain apolipoproteins plays a key role factor in the development of early atherosclerosis and in the elucidation of biochemical markers of the primary damages of cerebral vessels. PMID- 9752681 TI - [A new approach toward analysis of quality of inpatient psychiatric care]. AB - A new approach to assessment of quality of psychiatric hospital care is proposed. It is based on comparison of goals of hospitalisation and results achieved. The necessary condition for such assessment is formulation of these goals for each patient. Three components should be considered: clinical which coincides with the prognosis; social which includes achievement of the patient adaptation and psychological which assesses patient's satisfaction with the treatment. The preliminary results of practical use of the proposed method are presented. PMID- 9752682 TI - [The influence of oxygen baths on the condition of patients with discirculatory encephalopathy]. PMID- 9752683 TI - [Computer-tomographic basis for approaches to the assessment of post-stroke motor disorders]. PMID- 9752684 TI - [Contemporary achievements in diagnosis and prevention of mitochondrial diseases]. PMID- 9752685 TI - [Psychotropic drugs in somatic disorders]. PMID- 9752686 TI - [On the improvement of classification of cerebrovascular disorders]. PMID- 9752687 TI - [Relationship between glomerular epithelial cell injury and proteinuria in IgA nephropathy]. AB - The present study was designed to elucidate whether glomerular epithelial cell (GEC) injury is associated with the mechanism of proteinuria in biopsy-proven IgA nephropathy (IgAN). Twenty-four IgAN patients were divided into 4 groups based on their grade of proteinuria. Light microscopic examination revealed that GEC injury appeared prior to glomerular lesions such as sclerosis, crescent and adhesion. Reduced glomerular expression of C3b receptor (CR1), an indicator of GEC injury, was detected initially at the portion of damaged GEC and around the lesions of sclerosis, crescent and adhesion. CR1 expression eventually disappeared in the group IV patients who had nephrotic range proteinuria. Moreover, the grade of proteinuria in IgAN patients was associated with GEC injury and reduced glomerular expression of CR1. Taken together, GEC injury might be an important pathological finding which appears initially in the glomeruli, suggesting that GEC injury is a predictor of proteinuria in IgAN patients. PMID- 9752689 TI - [Severity of glomerular damage with IgA nephropathy and renal prognosis: a comparison between pattern classification and calculated classification]. AB - In our laboratory, a semi-quantitative pattern classification (PC) for the distribution pattern of glomerular sclerotic lesions in biopsied renal specimens with IgA nephropathy has been utilized, and found to be quite beneficial for predicting the patient's prognosis. In the present study conducted to re-evaluate this classification, 503 patients with IgA nephropathy (male/female, 256/247; mean age: 32.1 +/- 13.5 yrs; follow-up: 9.3 +/- 4.5 yrs) were used. The patients had been divided originally into 5 groups based on a previous PC: minimal, mild, moderate, severe and advanced groups. Their glomerular lesions were classified as mild, moderate, severe and global sclerosis, and were given scores of 1-4 points. The mean glomerular score was then calculated for each patient, as a value of the calculated classification (CC), and all patients were then re-divided into 5 groups based on their scores. The renal survival curves in the CC were similar to those in PC, and no significant differences in the renal survival rates were found between the classifications in each group, thus suggesting that the CC has a similar predictive power for renal survival. Although 7 of 39 cases (18%) with global sclerosis in PC groups 4 and 5 shifted down to CC group 3, of 61 patients with global sclerosis in PC group 1 who had a good prognosis, 26 cases (43%) shifted up to the CC group 2 and 6 cases (8%) changed to the CC group 3. As a result, the predictive power in patients in the lower CC groups was lost for the renal survival rate. In conclusion, statistical comparison between the PC and CC groups revealed that global sclerosis presents in non/minimally affected glomeruli as a nonspecific alteration. Severely advanced cases also possess a high incidence of globally sclerotic glomeruli (87%), and therefore the occurrence of global sclerosis may involve two different pathogenic mechanisms. PMID- 9752688 TI - [A clinicopathological study of recurrent IgA nephropathy following renal transplantation]. AB - The present study was conducted to examine the clinicopathological features of recurrent IgA nephropathy (IgAN) following renal transplantation. Serial renal biopsies were performed regularly at 0-hour, 1-hour and 2-hours, and 39 episode biopsies were carried out when patients had increased serum creatinine levels and proteinuria. In 49 renal allograft recipients with IgAN, 12 patients were proved to be recurrent IgAN (24.5%). There was a significantly increased five- and ten year risk of graft loss in the renal allograft recipients with biopsy-proved recurrent IgAN. Graft survival in 49 renal allograft recipients with IgAN was worse (68.8% at 5 years and 40.4% at 10 years) than that in 997 whole transplants (80.7% at 5 years, and 67.7% at 10 years). We found significant differences in the prevalence of HLA-DR4 (66.7%) and BW35 (25%) in the renal allograft recipients with recurrent IgAN when compared with normal healthy subjects. The renal allograft recipients with recurrent IgAN had a high incidence of proteinuria (8/12), hypertension (9/12) and renal dysfunction of less than 50 ml/min (7/12). Mean hemodialysis duration before renal transplantation in recurrent IgAN transplants was 12.5 months, which was shorter than in those without recurrent IgAN. Histopathological studies revealed that renal lesions due to IgAN frequently appeared in the renal allograft recipients with recurrent IgAN. Taken together, these findings suggest that donor-recipient matching may be carefully reconsidered, and recurrent IgAN after renal transplantation must be treated with effective immunosuppressive therapy. PMID- 9752690 TI - [Evaluation of the severity of glomerular damage in IgA nephropathy: significance of hyaline glomeruli in pattern classification]. AB - Hyaline glomeruli are observed frequently in biopsied samples with mild to severe glomerular damage. We thus investigated whether or not all hyaline glomeruli have the same semiquantitative values in order to predict accurately the renal prognosis. We histopathologically studied 503 patients with IgA nephropathy (256 males and 247 females; mean age: 32.1 +/- 13.5 yrs), whose prognoses were followed for more than 3 yrs (mean follow-up period: 9.3 +/- 4.5 yrs). Cases with severely damaged prolifero-sclerotic glomeruli showed poor prognoses (severely damaged glomeruli: (-) vs ( renal death rate: 6% vs 52%, p < 0.0001; the 15 year survival rate: 89% vs 52%, p < 0.0001) and hyaline glomeruli were seen in 68% of the cases. Hyaline glomeruli were observed in 29% of the cases with mild glomerular damage. However, the renal prognosis was not affected by the presence of hyaline glomeruli based on the analysis of the generalized Wilcoxon test. These results indicated that hyaline glomeruli express two different characteristics for renal prognosis. Therefore, the appearance of hyaline glomeruli alone is less important for the prediction of renal prognosis, but the appearance of both glomeruli with hyalinosis and those with severe damage was found useful in accurately predicting the renal prognosis. PMID- 9752691 TI - [Aldehyde dehydrogenase 2(ALDH2) gene polymorphism in NIDDM patients with chronic renal failure]. AB - In order to investigate the influence of aldehyde dehydrogenase 2(ALDH2) genotype in the pathogenesis of nephropathy due to non-insulin dependent diabetes mellitus (NIDDM), genotyping of ALDH2 was measured using the PCR-RFLP method in patients with NIDDM on chronic hemodialysis (HD). The results were as follows; 1) The frequency of active ALDH2 was 63% and that of inactive ALDH2 was 37%. 2) The percentage of active ALDH2 was significantly higher in patients with alcohol tolerance than that in those without it (38%). 3) The estimated amount of alcohol consumption in the past was 506 +/- 720 g/week in the active ALDH2 group, and 156 +/- 288 g/week in the inactive ALDH2 group, showing a significant difference between the two groups. 4) Interdialytic body weight gain was larger in patients with active ALDH2 than in those with inactive ALDH2. Since the frequency of active ALDH2 was similar to that in patients without nephropathy, these results do not support the hypothesis that ALDH2 gene polymorphism is involved in the development and persistence of chronic renal failure due to NIDDM. However, salt and water craving in dialysis patients may be influenced partially by an active ALDH2 gene. PMID- 9752692 TI - [A child case of idiopathic membranous glomerulonephritis associated with isolated anti-nuclear antibody detected by urine mass-screening of school children]. AB - A 12-year-old boy was referred to our hospital because of persistent hematuria with proteinuria. He was found to have urine abnormalities by a school mass screening and visited a hospital, where a routine examination revealed proteinuria of 190 mg/d/, hematuria of 2+ and anti-nuclear antibody (ANA) of 1: 320 with a speckled pattern. The other laboratory findings, including anti-DNA antibody were unremarkable. Hypocomplementemia was not seen. Although he had no disabilities and manifestations suggesting systemic lupus erythematosus, urine abnormalities persisted. Percutaneous renal biopsy findings demonstrated stage II diffuse membranous glomerulonephritis (MGN). Neither anti-hepatitis B virus antigens and antibodies nor hepatitis C virus antibody were detected in his sera. Thus, the diagnosis of idiopathic MGN was made. Initiation of a 6-month course of alternate-day prednisolone (an initial dosage at 30 mg) combined with an anti thrombocyte agent resulted in gradual subsidence of the urine abnormalities and ANA titer. Although pathogenesis of his MGN remains speculative, possible activation of autoimmunities, which led to ANA positivity, might be responsible. PMID- 9752693 TI - [A female case of Fournier's gangrene in a patient with lupus nephritis]. AB - Infection is one of the common causes of death in patients with systemic lupus erythematosus (SLE). It is associated with the use of immunosuppressive agents, renal failure, and increased disease activity. Fournier's gangrene is a necrotizing fasciitis occurring in the genital region. It is rare, but can be crucial if surgical drainage is delayed. We report a female case of Fournier's gangrene occurring in a patient with lupus nephritis and chronic renal failure. The patient was a 21-year-old female with chronic renal failure due to lupus nephritis. She had suffered from watery diarrhea one month before admission. It improved after increasing the dose of prednisolone, but, she was complicated with Bartholin abscess. The vaginal pain rapidly spread to the left lower quadrant abdomen despite treatment with oral cephalosporin. Focal incision was performed and black fluid emerged with a foul smell. Pelvic computed tomography (CT) revealed many bubbles in that region. She was found to have septic shock on transfer to our hospital. Thereafter, emergency debridement was performed, followed by antibiotic therapy and hyperbaric oxygen therapy. Organisms were found to be 5 anerobes, such as Bacteroides species, and 3 aerobics, such as Morganella morganii. Fournier's gangrene was improved via these treatments, but she needed maintenance hemodialysis. Fournier's gangrene complication should be considered in SLE with urogenital infection. PMID- 9752694 TI - [Immune complex-mediated glomerulonephritis associated with infective endocarditis with aortic valve vegetation]. AB - A 61-year-old male was referred to our hospital for rapidly progressive azotemia. He was also found to have huge vegetation at the aortic valve causing regurgitation. Biochemical examinations revealed the presence of an immunocomplex associated with decreased circulating complements. In biopsy samples from the kidney, we found the presence of fibrillar crescents, proliferation of mesangial cells, increase in extracellular matrix proteins, atrophy of tubules, infiltration of mononuclear cells in the interstitial regions, high density deposits in the mesangial area and mesangial interposition. Since the patient strongly rejected operative treatment by valvular replacement, we continued non invasive treatment such as hemodialysis and treatment with penicillin G. This transiently improved the condition of the patient, including biochemical data and cardiac function, but there was no reduction in the size of vegetation at the aortic valve and the bacteria responsible for infective endocarditis were not identified. About three months after admission, overt signs of congestive heart failure emerged and the patients was subjected to intensive care with a respirator and hemodynamic monitoring. Although the cardiac function was improved, concomitant severe pneumonia occurred and the patient died of septic shock. Thus, we report a rare case in whom immune complex-mediated glomerulonephritis was associated with infective endocarditis with aortic valve vegetation. PMID- 9752695 TI - [Role of gap junction in the stomach]. PMID- 9752696 TI - [Diagnosis and prognosis of reactive lymphoreticular hyperplasia (RLH) or mucosa associated lymphoid tissue (MALT) lymphoma]. AB - We studied the diagnostic significance of immunohistolochemical staining and immunoglobuline gene rearrangement of jumbo-biopsy specimen from 14 patients with stomach lesions which were difficult to distingwish between reactive lymphoreticular hyperplasia (RLH) and mucosa-associated lymphoid tissue lymphoma (MALT lymphoma). We also investigated Helicobacter pylori (HP) infection in the patients and followed their clinical courses from a mean observation period of 3 years and 4 months following initial diagnosis. As a result, 7 of the 14 cases were diagnosed as having MALT lymphoma. All of them were resected, and then the diagnosis was confirmed. Metastasis was found in 2 cases. The other 7 cases were diagnosed as RLH. A favorable prognosis during follow up without any treatment supported the belief that they were non-malignant lesions. HP infections were observed on 83% of the RLH cases and 57% of MALT lymphoma cases. In one of the case of RLH, the lesion disappeared after eradication of HP. PMID- 9752697 TI - [A case of gastric inflammatory fibroid polyp which caused ball valve syndrome]. PMID- 9752698 TI - [A case of mesenteric panniculitis complicated with malignant lymphoma]. PMID- 9752699 TI - [A case of minute IIa + IIc type early colonic cancer (5 mm in size and sm 1 in invasion depth) with metachronous liver metastasis]. PMID- 9752700 TI - [Perforated duodenal diverticulum caused by enterolith]. PMID- 9752701 TI - [Report of an autopsy cases of hepatocellular carcinoma with marked pulmonary hypertension due to multiple pulmonary thrombus]. PMID- 9752702 TI - [A case of adult chronic active EB virus infection associated with prolonged hepatitis after the occurrence of infectious mononucleosis]. PMID- 9752703 TI - [A case of idiopathic hemochromatosis which occurred in three siblings with high level of serum CA 19-9]. PMID- 9752704 TI - [A case of colonic hemorrhage caused by splenic artery pseudoaneurysm penetrating into the colon: management by transcatheter arterial embolization]. PMID- 9752705 TI - [A case of hypovascular acinar cell carcinoma of pancreas, liver metastatic lesions of which showed hypervascular]. PMID- 9752707 TI - "It just doesn't matter, it just doesn't matter, it just doesn't matter,...". PMID- 9752706 TI - [Intracystic hemorrhage of simple hepatic cyst--report of a case]. PMID- 9752708 TI - Global trends, needs, issues. AB - Worldwide, Pharmaceutical Plant Management struggles with the competing priorities of lowering costs, rising customer expectations, more demanding government regulations, and the need to reduce cycle times especially in the introduction of new products. All of this takes place in an environment of global competition, regulatory harmonization, mergers and downsizing, and employee insecurity. Employees are expected to do more with less, work with more sophisticated equipment and processes, take more personal responsibility for quality and productivity, work in teams, etc. In summary, we are talking about CHANGE, the speed of which will accelerate in the years to come. This presentation will discuss how some pharmaceutical plants are addressing these challenges. Examples will be given in the areas of validation, process reengineering, risk analysis, role of the quality function and people. It is my contention that most of the global trends today are insufficient to meet the challenges that we face. I hope that this presentation will generate some ideas on what the global trends should be. PMID- 9752709 TI - Origin of bacteria in the clean room and their growth requirements. PMID- 9752710 TI - Industrial manufacture of parenteral products in The Netherlands. A survey of eight years of media fills and sterility testing. AB - Sterility testing and media fills are essential requirements in the pharmaceutical industry. With the results obtained the manufacturer must ensure that the aseptic filling process is under control. In an eight year (1988-1995) retrospective survey of three major Dutch pharmaceutical companies the performance of the sterility test, of media fills and their relationship have been statistically evaluated. The products included human and veterinary pharmaceuticals and biologicals, and were divided into six different groups according to their production process and primary containers. A distinction was made between the results from the period 1985-1991 and the period 1992-1995, because this made the statistical analyses of a number of types of products possible, and because some significant changes in the production process were made in 1991 at some of the production sites. The results of the evaluation of the sterility test show that the frequency of false positives has not changed significantly. The overall rate of false positives of 0.17% is a factor ten better than considered acceptable by USP XXIII. For all product groups the frequency of positive sterility tests has decreased during the period of investigation. During the period 1992-1995 there was no significant difference between the results of product sterility tests and the negative controls for any of the product groups. This indicates that given the present state-of-the-art production the sterility test offers little or no additional security. The overall contamination of the media fills done in the more recent period is a factor twenty better than the limit given in the PDA Technical Monograph No. 2. In the more recent period there is a good agreement between the observed positive rate of the sterility tests and the positive rate that had been estimated from the results of the media fills. This indicates that despite some shortcomings, media fills are an adequate simulation of the production process and can be used to give an estimate of the rejection rate for the various product groups. The overall conclusion is that the production conditions of the participating Dutch pharmaceutical companies comply with the current international guidelines for aseptic production and sterility testing. PMID- 9752711 TI - Comparative mold and yeast recovery analysis (the effect of differing incubation temperature ranges and growth media). AB - Environmental monitoring methodology for recovering fungal organisms often dictates the use of a selective medium incubated at ambient temperatures (20-25 degrees C) for as many as seven days incubation to ensure reliable recovery. However, these methods (which must remain standardized for identification purposes) are not the only avenue environmental monitoring programs may follow. This study comparatively analyzed recovery rates of fungal organisms cultured on both a general purpose bacteriological nutrient medium (Tryptic Soy Agar supplemented with Lecithin and Polysorbate 80), and on a medium selective for the growth of yeasts and molds (Sabouraud Dextrose Agar). The bacteriologic medium was incubated at elevated temperatures (30-35 degrees C) for 70-72 hours then transferred to ambient temperatures for another 70-72 hours incubation. The control case selective medium was incubated solely at 20-25 degrees C for 110-130 hours. Additionally, in a separate test the selective medium was incubated at the same temperature and time specifications as the bacteriologic medium to analyze recovery capabilities. Equivalent or better recovery was obtained for all test panel organisms of yeasts and molds using Tryptic Soy Agar supplemented with Lecithin and Polysorbate 80 incubated at 30-35 degrees C. Equivalent or better recovery was obtained for eight of the nine test panel organisms of yeasts and molds incubated at elevated temperatures on Sabouraud Dextrose Agar versus recovery on Agar versus recovery on Sabouraud Dextrose Agar incubated solely at 20-25 degrees C. No inhibition of growth was observed at the elevated temperature range of 30-35 degrees C. Three days incubation at elevated temperatures was a sufficient incubation period to detect the test organisms cultured on Tryptic Soy Agar supplemented with Lecithin and Polysorbate 80. PMID- 9752712 TI - Emulsion formulations for intravenous administration of paclitaxel. PMID- 9752713 TI - "Absolute" sterility and "absolute" freedom from particle contamination. AB - Until the recent past, sterility of an injectable product was only discussed in absolute terms. Any description of sterility other than as an absolute could simply not be envisioned. While dealing in absolute yes/no statements is philosophically satisfying, these yes/no statements can't accommodate all real world scientific problems. Among these problems is the sterility problems faced in the mass production of injectable compounds. Many descriptions of procedures employed to achieve sterility in parenteral production batches were reported in the literature. The theoretical framework that could unite the widespread observations and practices into practical methodology was missing until recently. Production line control of the sterility of injectable products was essentially based on gut evaluations. The present achievement of rational, production line control of product sterility is based on the recognition that product sterility could not be simply regarded as a sharply edged yes/no affair. The present rational control is based on the fact that the sterility of a product is determined by the degree of contamination in the product prior to sterilization and to the parameters of the sterilization process. The end result of the sterilization process is now described as a probabalistic reduction of the initial contamination. The essential laboratory measurements on which this conclusion was based is due to Pflug (1-3). He assembled a theoretical framework, based on experimental data, that characterizes the sterility achieved in an injectable product with a single number. The end result of the sterilization process is now described as a probabalistic reduction of the initial contamination. As in many disciplines, the ability to achieve an objective evaluation of this important attribute provided the basis for scientific analysis, improved control and thus improved production and reduced cost. An equivalent framework is essential for the communication and standardization of the results of a visual inspection for contaminating particles. The control of particle contamination in injectable products is a two-fold problem for the pharmaceutical industry. The two parts of the problem are 1) achieving contamination free product and 2) achieving this contamination free quality at an economic cost acceptable to the user. Today, there is no commonly accepted framework for the definition or analysis of the results of a manual inspection for "visible" particles. Any progress toward global harmonization of the results of a visual particle inspection must commence with the development of a common scientific language with which inspection security and economic effectiveness of an inspection can be discussed and rationally evaluated. The tools with which to define and control the results of this inspection have been developed in biophysics, illumination engineering, optics, pharmaceutical manufacturing and statistics. With them, statistically replicable measures have been developed. The statistically replicable measures are then used to evaluate the results of semi- and fully automated particle inspection systems in terms of human inspection performance. The numerical evaluation of both the freedom from contamination achieved with an inspection for particle contamination and the economic effectiveness of the inspection are compared to Pflug's sterility index. In the case of particle contamination, the final product quality depends on product quality prior to inspection and to the parameters of the inspection process. PMID- 9752714 TI - The pressure hold/drop integrity test; its correlation to diffusive flow. AB - The pressure-drop/hold procedure enables the diffusive flow integrity testing of filters to be performed without breaching the system downstream of the filter. It is not necessary to measure volumetrically the diffused gas on the downstream side of the filter. By means of pressure transducers the pressure loss is determined upstream; thus eliminating the threat of sepsis due to down-stream invasions. The pressure decay exercise can be used to characterize the various filter types. A constancy of filter manufacture is required for a given filter type. Unless the pressure drop exceeds the value established as the maximum allowable decay, the filter is judged to be integral. It qualifies as a sterilizing grade filter. Excessive pressure decays will also eventuate from leaks, as from improperly sealed housings. Performed prior to the filtration, the procedure serves to eliminate the wasteful use of an imperfect system, whether caused by faulty sealing, incorrect filter type or flawed filters. Where leaks are detected, the filter can be reexamined for its integrity. To enable the pressure-drop procedure to serve as an integrity test, the measured pressure decays require being correlated with organism retention data. This is made possible by the arithmetic conversion of the pressure decay curve into the conventional diffusive airflow curve established to have such a correlation. The transformation of the pressure-drop curve into the differential airflow plot is automatically performed by certain of the automated integrity test machines. These devices, utilizing pressure transducers, are capable of measuring small pressure drops with requisite sensitivity; gauges commonly are not. Moreover, as previously stated, the measurements are advantageously made on the upside of the filter. The use of automated test machines is, therefore, recommended for the performance of the pressure hold/drop integrity test in furtherance of the practice of filter integrity testing. PMID- 9752715 TI - PDA comments: reporting of errors and accidents. Parenteral Drug Association. PMID- 9752716 TI - Pharmaceutical Package Integrity. Parenteral Drug Association. PMID- 9752717 TI - Neurofilaments in health and disease. AB - This article reviews current knowledge of neurofilament structure, phosphorylation, and function and neurofilament involvement in disease. Neurofilaments are obligate heteropolymers requiring the NF-L subunit together with either the NF-M or the NF-H subunit for polymer formation. Neurofilaments are very dynamic structures; they contain phosphorylation sites for a large number of protein kinases, including protein kinase A (PKA), protein kinase C (PKC), cyclin-dependent kinase 5 (Cdk5), extracellular signal regulated kinase (ERK), glycogen synthase kinase-3 (GSK-3), and stress-activated protein kinase gamma (SAPK gamma). Most of the neurofilament phosphorylation sites, located in tail regions of NF-M and NF-H, consist of the repeat sequence motif, Lys-Ser-Pro (KSP). In addition to the well-established role of neurofilaments in the control of axon caliber, there is growing evidence based on transgenic mouse studies that neurofilaments can affect the dynamics and perhaps the function of other cytoskeletal elements, such as microtubules and actin filaments. Perturbations in phosphorylation or in metabolism of neurofilaments are frequently observed in neurodegenerative diseases. A down-regulation of mRNA encoding neurofilament proteins and the presence of neurofilament deposits are common features of human neurodegenerative diseases, including amyotrophic lateral sclerosis (ALS), Parkinson's disease, and Alzheimer's disease. Although the extent to which neurofilament abnormalities contribute to pathogenesis in these human diseases remains unknown, emerging evidence, based primarily on transgenic mouse studies and on the discovery of deletion mutations in the NF-H gene of some ALS eases, suggests that disorganized neurofilaments can provoke selective degeneration and death of neurons. An interference of axonal transport by disorganized neurofilaments has been proposed as one possible mechanism of neurofilament induced pathology. Other factors that can potentially lead to the accumulation of neurofilaments will be discussed as well as the emerging evidence for neurofilaments as being possible targets of oxidative damage by mutations in the superoxide dismutase enzyme (SOD1); such mutations are responsible for approximately 20% of familial ALS cases. PMID- 9752718 TI - Regulation of cytochrome P450 gene transcription by phenobarbital. AB - The ability of phenobarbital to induce levels of drug metabolism in mammals has been known for over 40 years. However, the molecular mechanisms underlying increased expression of the genes of the key enzyme in drug metabolism, cytochrome P450, have not been elucidated, primarily because in vitro model systems in which the induction could be studied were not available. Transfected primary cultured hepatocytes, transfection of liver in situ, and transgenic mice now provide suitable models for phenobarbital induction. In this review, progress toward understanding the mechanism of phenobarbital induction of gene expression is discussed with an emphasis on the mammalian genes, CYP2B1, CYP2B2, and Cyp2b10, which are most highly inducible by phenobarbital. Barbiturate induction of P450s in Bacillus megaterium, which is the system best understood, and its relevance to mammalian mechanisms of induction are also discussed. In B. megaterium, the binding of a repressor to several motifs is reversed by direct effects of barbiturates and by induction of positively acting factors. One of the repressor binding sites, the barbie box, is present in many mammalian phenobarbital-inducible genes, including the promimal promoter regions of CYP2B1, CYP2B2, and Cyp2B10. In the mammalian P450 genes, evidence has been proposed for phenobarbital-regulated elements both in the proximal promoter region and in a distal enhancer region. The role of the proximal region is controversial. A positively acting element that overlaps the barbie box sequence and a negative element have been proposed to mediate induction of CYP2B1/2, based primarily on protein binding and cell-free transcription assays. In contrast, other investigators have not found differences in phenobarbital-dependent protein binding in the proximal promoter region nor mediation of phenobarbital induction by this region. A distal gene fragment, at about -2000 kb in CYP2B1, CYP2B2, and Cyp2b10, has been shown to be a phenobarbital-responsive enhancer independent of proximal promoter elements. This fragment contains several binding sites for proteins and several functional elements, including an NF-1 site, and, therefore, has been designated as a phenobarbital-responsive unit. Possible models are presented in which phenobarbital treatment induces altered chromatin structure, which allows the binding of positively acting factors, or activates factors already bound, to the distal enhancer and the proximal promoter. PMID- 9752719 TI - RNA and protein interactions modulated by protein arginine methylation. AB - This review summarizes the current status of protein arginine N-methylation reactions. These covalent modifications of proteins are now recognized in a number of eukaryotic proteins and their functional significance is beginning to be understood. Genes that encode those methyltransferases specific for catalyzing the formation of asymmetric dimethylarginine have been identified. The enzyme modifies a number of generally nuclear or nucleolar proteins that interact with nucleic acids, particularly RNA. Postulated roles for these reactions include signal transduction, nuclear transport, or a direct modulation of nucleic acid interactions. A second methyltransferase activity that symmetrically dimethylates an arginine residue in myelin basic protein, a major component of the axon sheath, has also been characterized. However, a gene encoding this activity has not been identified to date and the cellular function for this methylation reaction has not been clearly established. From the analysis of the sequences surrounding known arginine methylation sites, we have determined consensus methyl accepting sequences that may be useful in identifying novel substrates for these enzymes and may shed further light on their physiological role. PMID- 9752720 TI - Genetic regulation of phospholipid metabolism: yeast as a model eukaryote. AB - Baker's yeast, Saccharomyces cerevisiae, is an excellent and an increasingly important model for the study of fundamental questions in eukaryotic cell biology and genetic regulation. The fission yeast, Schizosaccharomyces pombe, although not as intensively studied as S. cerevisiae, also has many advantages as a model system. In this review, we discuss progress over the past several decades in biochemical and molecular genetic studies of the regulation of phospholipid metabolism in these two organisms and higher eukaryotes. In S. cerevisiae, following the recent completion of the yeast genome project, a very high percentage of the gene-enzyme relationships in phospholipid metabolism have been assigned and the remaining assignments are expected to be completed rapidly. Complex transcriptional regulation, sensitive to the availability of phospholipid precusors, as well as growth phase, coordinates the expression of the structural genes encoding these enzymes in S. cerevisiae. In this article, this regulation is described, the mechanism by which the cell senses the ongoing metabolic activity in the pathways for phospholipid biosynthesis is discussed, and a model is presented. Recent information relating to the role of phosphatidylcholine turnover in S. cerevisiae and its relationship to the secretory pathway, as well as to the regulation of phospholipid metabolism, is also presented. Similarities in the role of phospholipase D-mediated phosphatidylcholine turnover in the secretory process in yeast and mammals lend further credence to yeast as a model system. PMID- 9752721 TI - Inosine-5'-monophosphate dehydrogenase: regulation of expression and role in cellular proliferation and T lymphocyte activation. AB - Guanine nucleotide synthesis is essential for the maintenance of normal cell growth and function, as well as for cellular transformation and immune responses. The expression of two genes encoding human inosine-5'-monophosphate dehyrogenase (IMPDH) type I and type II results in the translation of catalytically indistinguishable enzymes that control the rate-limiting step in the de novo synthesis of guanine nucleotides. Cellular IMPDH activity is increased more than 10-fold in activated peripheral blood T lymphocytes and is attributable to the increased expression of both the type I and type II enzymes. In contrast, abrogation of cellular IMPDH activity by selective inhibitors prevents T lymphocyte activation and establishes a requirement for elevated IMPDH activity in T lymphocytic responses. In order to assess the molecular mechanisms governing the expression of the IMPDH type I and type II genes in resting and activated peripheral blood T lymphocytes, we have cloned the human IMPDH type I and type II genes and characterized their genomic organization and their respective 5' flanking regions. Both genes contain 14 highly conserved exons that vary in size from 49 to 207 base pairs. However, the intron structures are completely divergent, resulting in disparities in gene length (18 kilobases for type I and 5.8 kilobases for type II). In addition, the 5'-regulatory sequences are highly divergent; expression of the IMPDH type I gene is controlled by three distinct promoters in a tissue specific manner while the type II gene is regulated by a single promoter and closely flanked in the 5' region by a gene of unknown function. The conservation of the IMPDH type I and type II coding sequence in the presence of highly divergent 5'-regulatory sequences points to a multifactorial control of enzyme expression and suggests that tissue-specific and/or developmentally specific regulation of expression may be important. Delineation of these regulatory mechanisms will aid in the elucidation of the signaling events that ultimately lead to the synthesis of guanine nucleotides required for cellular entry into S phase and the initiation of DNA replication. PMID- 9752722 TI - Structure and function analysis of Pseudomonas plant cell wall hydrolases. AB - Hydrolysis of the major structural polysaccharides of plant cell walls by the aerobic soil bacterium Pseudomonas fluorescens subsp. cellulosa is attributable to the production of multiple extracellular cellulase and hemicellulase enzymes, which are the products of distinct genes belonging to multigene families. Cloning and sequencing of individual genes, coupled with gene sectioning and functional analysis of the encoded proteins have provided a detailed picture of structure/function relationships and have established the cellulase-hemicellulase system of P. fluorescens subsp. cellulosa as a model for the plant cell wall degrading enzyme systems of aerobic cellulolytic bacteria. Cellulose- and xylan degrading enzymes produced by the pseudomonad are typically modular in structure and contain catalytic and noncatalytic domains joined together by serine-rich linker sequences. The cellulases include a cellodextrinase; a beta-glucan glucohydrolase and multiple endoglucanases, containing catalytic domains belonging to glycosyl hydrolase families 5, 9, and 45; and cellulose-binding domains of families II and X, both of which are present in each enzyme. Endo acting xylanases, with catalytic domains belonging to families 10 and 11, and accessory xylan-degrading enzymes produced by P. fluorescens subsp. cellulosa contain cellulose-binding domains of families II, X, and XI, which act by promoting close contact between the catalytic domain of the enzyme and its target substrate. A domain homologous with NodB from rhizobia, present in one xylanase, functions as a deacetylase. Mananase, arabinanase, and galactanase produced by the pseudomonad are single domain enzymes. Crystallographic studies, coupled with detailed kinetic analysis of mutant forms of the enzyme in which key residues have been altered by site-directed mutagenesis, have shown that xylanase A (family 10) has 8-fold alpha/beta barrel architecture, an extended substrate binding cleft containing at least six xylose-binding pockets and a calcium binding site that protects the enzyme from thermal inactivation, thermal unfolding, and attack by proteinases. Kinetic studies of mutant and wild-type forms of a mannanase and a galactanase from P. fluorescens subsp. cellulosa have enabled the catalytic mechanisms and key catalytic residues of these enzymes to be identified. PMID- 9752723 TI - Regulation of the spatiotemporal pattern of expression of the glutamine synthetase gene. AB - Glutamine synthetase, the enzyme that catalyzes the ATP-dependent conversion of glutamate and ammonia into glutamine, is expressed in a tissue-specific and developmentally controlled manner. The first part of this review focuses on its spatiotemporal pattern of expression, the factors that regulate its levels under (patho)physiological conditions, and its role in glutamine, glutamate, and ammonia metabolism in mammals. Glutamine synthetase protein stability is more than 10-fold reduced by its product glutamine and by covalent modifications. During late fetal development, translational efficiency increases more than 10 fold. Glutamine synthetase mRNA stability is negatively affected by cAMP, whereas glucocorticoids, growth hormone, insulin (all positive), and cAMP (negative) regulate its rate of transcription. The signal transduction pathways by which these factors may regulate the expression of glutamine synthetase are briefly discussed. The second part of the review focuses on the evolution, structure, and transcriptional regulation of the glutamine synthetase gene in rat and chicken. Two enhancers (at -6.5 and -2.5 kb) were identified in the upstream region and two enhancers (between +156 and +857 bp) in the first intron of the rat glutamine synthetase gene. In addition, sequence analysis suggests a regulatory role for regions in the 3' untranslated region of the gene. The immediate-upstream region of the chicken glutamine synthetase gene is responsible for its cell-specific expression, whereas the glucocorticoid-induced developmental appearance in the neural retina is governed by its far-upstream region. PMID- 9752725 TI - Control of meiotic recombination in Schizosaccharomyces pombe. AB - Homologous recombination occurs at high frequency during meiosis and is essential for the proper segregation of chromosomes and the generation of genetic diversity. Meiotic recombination is controlled in numerous ways. In the fission yeast Schizosaccharomyces pombe nutritional starvation induces meiosis and high level expression of many genes, including numerous recombination (rec) genes, whose products are required for recombination. Accompanying the two meiotic divisions are profound changes in nuclear and chromosomal structure and movement, which may play an important role in meiotic recombination. Although recombination occurs throughout the genome, it occurs at high frequency in some intervals (hotspots) and at low frequency in others (coldspots). The well-characterized hotspot M26 is activated by the Mts1/Mts2 protein; this site and its binding proteins interact with the local chromosomal structure to enhance recombination. A coldspot between the silent mating-type loci is repressed by identified proteins, which may also alter local chromatin. We discuss in detail the rec genes and the possible functions of their products, some but not all of which share homology with other identified proteins. Although some of the rec gene products are required for recombination throughout the genome, others demonstrate regional specificity and are required in certain genomic regions but not in others. Throughout the review contrasts are made with meiotic recombination in the more thoroughly studied budding yeast Saccharomyces cerevisiae. PMID- 9752724 TI - Structural organization and transcription regulation of nuclear genes encoding the mammalian cytochrome c oxidase complex. AB - Cytochrome c Oxidase (COX) is the terminal component of the bacterial as well as the mitochondrial respiratory chain complex that catalyzes the conversion of redox energy to ATP. In eukaryotes, the oligomeric enzyme is bound to mitochondrial innermembrane with subunits ranging from 7 to 13. Thus, its biosynthesis involves a coordinate interplay between nuclear and mitochondrial genomes. The largest subunits, I, II, and III, which represent the catalytic core of the enzyme, are encoded by the mitochondrial DNA and are synthesized within the mitochondria. The rest of the smaller subunits implicated in the regulatory function are encoded on the nuclear DNA and imported into mitochondria following their synthesis in the cytosol. Some of the nuclear coded subunits are expressed in tissue and developmental specific isologs. The ubiquitous subunits IV, Va, Vb, VIb, VIc, VIIb, VIIc, and VIII (L) are detected in all the tissues, although the mRNA levels for the individual subunits vary in different tissues. The tissue specific isologs VIa (H), VIIa (H), and VIII (H) are exclusive to heart and skeletal muscle. cDNA sequence analysis of nuclear coded subunits reveals 60 to 90% conservation among species both at the amino acid and nucleotide level, with the exception of subunit VIII, which exhibits 40 to 80% interspecies homology. Functional genes for COX subunits IV, Vb, VIa 'L' & 'H', VIIa 'L' & 'H', VIIc and VIII (H) from different mammalian species and their 5' flanking putative promoter regions have been sequenced and extensively characterized. The size of the genes range from 2 to 10 kb in length. Although the number of introns and exons are identical between different species for a given gene, the size varies across the species. A majority of COX genes investigated, with the exception of muscle specific COXVIII(H) gene, lack the canonical 'TATAA' sequence and contain GC-rich sequences at the immediate upstream region of transcription start site(s). In this respect, the promoter structure of COX genes resemble those of many house keeping genes. The ubiquitous COX genes show extensive 5' heterogeneity with multiple transcription initiation sites that bind to both general as well as specialized transcription factors such as YY1 and GABP (NRF2/ets). The transcription activity of the promoter in most of the ubiquitous genes is regulated by factors binding to the 5' upstream Sp1, NRF1, GABP (NRF2), and YY1 sites. Additionally, the murine COXVb promoter contains a negative regulatory region that encompasses the binding motifs with partial or full consensus to YY1, GTG, CArG, and ets. Interestingly, the muscle-specific COX genes contain a number of striated muscle-specific regulatory motifs such as E box, CArG, and MEF2 at the proximal promoter regions. While the regulation of COXVIa (H) gene involves factors binding to both MEF2 and E box in a skeletal muscle-specific fashion, the COXVIII (H) gene is regulated by factors binding to two tandomly duplicated E boxes in both skeletal and cardiac myocytes. The cardiac-specific factor has been suggested to be a novel bHLH protein. Mammalian COX genes provide a valuable system to study mechanisms of coordinated regulation of nuclear and mitochondrial genes. The presence of conserved sequence motifs common to several of the nuclear genes, which encode mitochondrial proteins, suggest a possible regulatory function by common physiological factors like heme/O2/carbon source. Thus, a well orchestrated regulatory control and cross talks between the nuclear and mitochondrial genomes in response to changes in the mitochondrial metabolic conditions are key factors in the overall regulation of mitochondrial biogenesis. PMID- 9752726 TI - The nucleosome: a powerful regulator of transcription. AB - Nucleosomes provide the architectural framework for transcription. Histones, DNA elements, and transcription factors are organized into precise regulatory complexes. Positioned nucleosomes can facilitate or impede the transcription process. These structures are dynamic, reflecting the capacity of chromatin to adopt different functional states. Histones are mobile with respect to DNA sequence. Individual histone domains are targeted for posttranslational modifications. Histone acetylation promotes transcription factor access to nucleosomal DNA and relieves inhibitory effects on transcriptional initiation and elongation. The nucleosomal infrastructure emerges as powerful contributor to the regulation of gene activity. PMID- 9752727 TI - [The tissue reaction to acrylic plastics modified by supercritical extraction with carbon dioxide]. AB - The process of extraction of admixtures from acryl plastic widely used in dentistry by means of supercritical carbon dioxide (sc-CO2) was studied and effects of extraction conditions on biocompatibility and toxicity of resultant materials assessed, sc-CO2 effectively purified the specimens from toxic compounds (monomers and low-molecular oligomers, methylmethacrylate, dichloroethane) and notably improved the biocompatability of polymer implants. Tissue reaction to ethacryl and protacryl depends on the degree of implant polymerization and duration of extraction of toxic substances from polymer. PMID- 9752728 TI - [The biomedical efficacy of 2 variants of polyacrylamide gel- and hydroxyapatite based composite materials in the plastic repair of bone defects (experimental morphological research)]. AB - Tissue status at the site of experimental bone defects filled by two compositions based on polyacrylamide gel and hydroxyapatite with (experimental group) and without (controls) lysozyme was studied in rats by the histomorphological method. "Negative" symptoms, such as inflammation, formation of osteocyte-free bone at the interface of the defect, and reduction of red bone marrow were more manifest in the controls than in animals treated with lysozyme. In the test group substitution of composite material for connective tissue structures and bone reparation were much more active and rapid than in the controls. Inflammation and dystrophic changes at the interface of defects were less pronounced and gradually resolved. PMID- 9752729 TI - [The clinical evaluation of the quality of tooth filling after exposure to hydroxyethylidene diphosphonic acid]. AB - Ultrastructure of dental enamel was examined under electron microscope after exposure to hydroxyethylene diphosphonic (HEDP) acid and prophilometry of enamel surface was carried out after its exposure to acid salts; clinical studies in 105 patients aged 18-50 years with different diseases of hard dental tissues were carried out in order to assess the duration of filling preservation after dental enamel treatment with HEDP salts. Clinical results were assessed from subjective sensations of patients and data of examination. The criteria of assessing the results were presence of a filling in a filled carious cavity, marginal adhesion of the filling, alteration of color along the external interface of the filling, caries relapses, status of dental pulp and periapical tissues. Clinical and laboratory studies showed that treatment with HEDP acid creates favorable conditions for tooth filling. PMID- 9752731 TI - [The prevention of inflammatory complications in mandibular fractures by using infrared laser and magnetic-laser radiation]. AB - A total of 102 patients with mandibular fractures were treated by multiple modality treatment including infrared (IR) laser exposure and magnetic and laser therapy (MT). For monitoring the treatment efficacy and predicting its results, nonspecific defense factors and intensity of free-radical oxidation (FRO) in the saliva were assessed. IR laser and MT by the Ulei-2K device stimulated local defense factors, decreased the intensity of salivary FRO, and thus promoted the healing of mandibular fractures. PMID- 9752730 TI - [A comparative evaluation of modern antibacterial preparations in the treatment of a severe degree of periodontitis at a stage of exacerbation]. AB - Sensitivities of peptostreptococci, streptococci, Actinomyces, bacteroid, and fusobacterial strains pathogenic for the periodontium to wide-spectrum penicillines, cephalosporines, lincomycin, macrolides, metronidasole, and nitasole are compared. New macrolide antibiotics rulide. Macropene, gramicidin C, levomycetin, and rifampicin are highly effective. Some narrow-spectrum drugs, e.g. augmentin, cephalexin, and vancomycin (towards actinomycetes) were highly effective, too. PMID- 9752732 TI - [The clinical x-ray aspects of osteoplasty after osteotomy of the maxillary complex]. AB - Forty-six patients with combined deformations of the jaws were subjected to surgical treatment. Formalin-treated bone allotransplants together with wire splints were found to play the retention and fixing role for a year after surgery. During this period complete consolidation of osteotomied fragment of the maxillary complex takes place. No inflammations at the site of intervention were observed during the immediate and late periods after the operation. Formalin treated bone allograft formed from tibial or femoral bone is resistant to infection. PMID- 9752733 TI - [The strategy and planning of dental prophylaxis under the conditions of a transitional economy]. PMID- 9752734 TI - [The dynamics of caries intensity during the hygienic instruction and specific prophylaxis of children in central Yakutia]. AB - There was held the comparative evaluation of the influence of the mouth cavity hygiene and specific prophylactic measures on caries intensity of permanent teeth in 7-9-years-old children in the central Yakutia. The significant cariostatic effect was got in children of basic group with hygienic upbringing and specific prophylactics. So the complex use of hygienic upbringing and specific prophylactics (the systematic rinsing of mouth cavity with natrium fluoride according to methods of WHO) can be the effective measure of reducing the caries of permanent teeth in children of Yakutia. PMID- 9752735 TI - [Primary anodontia in X-linked hidrotic ectodermal dysplasia]. AB - Nine patients with congenital deciduous and permanent adentia are described for the first time in Russia. Published reports are reviewed and recommendations on the DNA and prenatal diagnosis offered, which permit detecting women at a high risk of giving birth to handicapped children. PMID- 9752736 TI - [The combined treatment of children with congenital and acquired mandibular defects and deformities]. AB - A total of 280 children and adolescents with congenital and acquired mandibular defects and deformations were treated. The main principle in the treatment of such children is to begin treatment as early as possible. Stages of rehabilitation and scope of therapeutic measures for each stage are determined. The best results were attained in patients administered multiple-modality treatment by surgical, orthodontic, physiotherapeutic methods and therapeutic exercises at an early age immediately after the defect was diagnosed. PMID- 9752737 TI - [Acid-base equilibrium in the oral cavity of children with congenital clefts of the upper lip and palate]. AB - Acid base balance (ABB) of the oral cavity is compared in children with cleft lip and palate before uranoplasty and 2-3 months and 1 year after it. A decompensated shift of ABB towards alkalosis before treatment was concomitant with transposition of the typical zones of bacterial shifting of ABB towards acidosis and alkalosis. ABB did not rapidly normalize after repair of the congenital defect. Preventive measures are recommended, aimed at removal or alleviation of alkalosis and retaining the adaptation reserve of the mechanisms regulating the ABB in the oral cavity, to be used both before and after surgery. PMID- 9752738 TI - [Dental care for patients with hematological diseases]. AB - Clinical and cytological studies in 41 hematological patients aged 19-70 years showed appreciable shifts in the immune status and nonspecific reactivity virtually in all examinees. Layers of cellular epithelium of late (V-VI) and early (III-IV) stages were more often detected in smears impressions than individual cells. Cells of stages I-II were scarce and never formed layers. In patients with leukemia, signs of dissociation of epitheliocyte nucleus and cytoplasm maturation were observed ("mature" cytoplasm and "young" nuclei). These specific shifts should be borne in mind when rendering dental care to hematological patients. PMID- 9752739 TI - [Medical informatics on the eve of the 21st century and the tasks of the dental service in Russia]. AB - The problems and tasks of information provision of dental service in Russia for all levels, from federal to regional health center are discussed as well as the concept of development and program of information provision in dentistry, software for automated working places of dentists, local network for dental centers and dentistry departments, software for differential diagnosis and treatment of the main dental diseases, such as caries, pulpitis, periodontitis, diseases of the periodontium and buccal mucosa, benign tumors, etc., for rapid screening of dental patients for AIDS and other diseases, design and software for computers intended for computer training of students and dentists, stomatology search and reference information system, etc. PMID- 9752740 TI - [The levels of decision making and execution and their role in the organization of a dental service]. PMID- 9752741 TI - [The prevalence of diseases of the oral mucosa in workers in the cotton processing industry]. AB - The prevalence [correction of incidence] of diseases of the buccal mucosa is assessed from records on dental status of 494 subjects engaged in cotton industry. A high prevalence [correction of incidence] of oral diseases is revealed. Candidiasis of the oral cavity and diseases of the lips are referred to occupational in this category of patients. PMID- 9752742 TI - [An analysis of the efficacy of orthodontic care for the population of Moscow based on data from an expert commission on quality]. PMID- 9752744 TI - [The 60th anniversary of the Department of Hospital Operative Dentistry and Maxillofacial Surgery of the Moscow Medical Stomatological Institute]. PMID- 9752745 TI - [Indebted for all to his native land, living only for his native land. Ivan Fedorovich Bush]. PMID- 9752746 TI - [Experience in conducting course exams on assignments in test form]. AB - Analysis of the results of examinations in oral surgery making use of tests in 319 fourth-year students showed that such a method of assessing students' knowledge is highly objective. PMID- 9752747 TI - Hepatitis 'A' virus epidemiology is changing in India. PMID- 9752748 TI - Management of pancreatic necrosis. PMID- 9752749 TI - High fidelity vertical transmission of genetic information: DNA replication and repair. PMID- 9752750 TI - Use of organic nitrates in gastroenterology. PMID- 9752751 TI - Biliary tract neoplasms. AB - From January 1993 to December 1995, complete records of patients with biliary neoplasms were analysed. A total of 124 patients were registered. Majority of patients were in the age range of 40-60 years (median 54 years). There were 38 males and 86 females. Histopathologically, adenocarcinoma was the commonest type (59%). Pain, jaundice and lump were noticed in 119, 54 and 77 patients respectively. Fifty six patients had associated gall stones. Ninety patients had metastatic disease at presentation. Majority of them (110/124) had advanced, inoperable disease and therefore were considered for palliative treatment. Only 14 patients (12%) were considered for curative treatment. Of these 14 patients, all the cases underwent surgery, 10 received radiotherapy and 10 received chemotherapy. Follow up was very poor. The survival of 14 patients, who received curative treatment, ranged from 2 months to 44 months with mean of 16 months. PMID- 9752752 TI - Typing and antibiotic susceptibility patterns of Vibrio cholerae during six consecutive cholera seasons in north India. AB - A total of 10,427 diarrhoeal stool specimens were cultured for Vibrio cholerae between 1992 and 1997. The isolation rates were 2%, 2.6%, 6.7%, 7.08%, 0.9% and 2.6% in the years from 1992 to 1997 respectively. Till 1992, Vibrio cholerae 01 ogawa was the predominant strain. In 1993, 81.3% of the isolates were of 0139 Bengal strain and the rest were V. cholerae 01. From 1994 to 1997, V. cholerae 01 ogawa was the predominant strain and there were no isolation of 0139 strain. The predominant phage type in 1992 and 1993 were T2 and T27 thereafter. Most Vibrio cholerae strains were sensitive to tetracycline, gentamycin, netromycin, norfloxacin and furazolidine. Strains were resistant to cotrimaxozole till 1996, but were 100% sensitive in 1997. Strains were sensitive to chloramphenicol till 1993 but acquired resistance thereafter. PMID- 9752753 TI - Autoimmune hepatitis in a patient with primary Sjogren's syndrome and selective IgA deficiency. PMID- 9752754 TI - Bilobed gallbladder with gallstones and choledocholithiasis. PMID- 9752755 TI - Leiomyosarcomas of the duodenum. PMID- 9752756 TI - Clinical course and management of pancreatoblastoma in children. AB - Pancreatoblastoma is a rare malignant tumour. Two children with this tumour were managed in the last 2 years. Both presented with progressively increasing abdominal mass. The diagnosis was established only after laparotomy. In the first child, an 8 year old girl, the mass was arising from the body of the pancreas and only incomplete resection was feasible. She received postoperative chemotherapy and went into remission for a few months before presenting with jaundice and abdominal pain due to recurrent, metastatic disease in the liver and porta hepatitis. Further therapy was refused by the patient because of anorexia and social problems. The second patient, a 5-year-old girl, underwent distal pancreatectomy for complete removal of a large mass arising from the tail of the pancreas. Chemotherapy was begun postoperatively but discontinued by the patient. However, she has remained disease free 1 year after diagnosis. Histologic, histochemical and ultrastructural features of the tumour are detailed and the management discussed. PMID- 9752757 TI - Profile of lower gastrointestinal bleeding in children from a tropical country. AB - Eighty five children were evaluated endoscopically for recurrent lower gastrointestinal (GI) bleeding. The male: female ratio was 2.4:1 with a mean age of 6 years (range 8 months to 2 years). After adequate bowel preparation endoscopic evaluation was done using olympus CF 101 colonoscope. Sedation was given only in two patients. Full length colonoscopy had been done in 16 cases only, to look for extent of disease in 8 cases and to ascertain site of bleeding when no lesion could be seen on sigmoidoscopy. Juvenile polyps were seen in 40 cases, amoebic ulcer in 20, solitary rectal ulcer in 4 and polyposis syndrome in 5 cases. Sigmoidoscopy alone could establish the diagnose in 76 cases. We conclude that flexible sigmoidoscopy alone is safe and adequate in ascertaining the cause of prolonged recurrent lower GI bleeding. PMID- 9752758 TI - Pitfalls in the management of Mirizzi's syndrome. AB - Pressure on the common hepatic duct due to a gallstone impacted in Hartmann's pouch or cystic duct results in jaundice and cholangitis. Repeated episodes of inflammation and pressure necrosis lead to the formation of a cholecysto choledochal fistula (Mirizzi's syndrome Type I & II). Preoperative diagnosis is difficult and a formal cholecystectomy may lead to bile duct injury. Of the 792 patients operated upon for symptomatic gallstone disease from June 1992 to June 1997 at our centre, 18 patients (2%) had Mirizzi's syndrome. There were 11 females and 5 males, with a mean age of 48 (SD 20; range 20-74) years. Thirteen patients (81%) presented with cholangitis. Ultrasound scan suggested the diagnosis of carcinoma gallbladder in 9 (56%). Endoscopic Retrograde Cholangiopancreatography (ERCP) confirmed the diagnosis in 16. Cholecystectomy was done by the fundus first technique. A complete cholecystectomy was done only if there was no cholecysto-choledochal fistula (n = 5), otherwise a cuff of gallbladder was used to repair the bile duct (n = 10). Hepatico-jejunostomy was done to drain the fistula in one patient. A T-tube drain was placed in the common bile duct (CBD) and a cholangiogram done, before closing the abdomen in all. Histology revealed carcinoma in fundus of gallbladder in one patient (6%). One patient died of haemobilia 3 weeks after operation. Wound infection developed in 5 (30%) patients and 12 (75%) have been followed up for a median period of 28 months. One patient developed a biliary stricture with intrahepatic stones and later underwent a hepatico-jejunostomy. Two have undergone repair of incisional hernia. High index of clinical suspicion, ERCP to clinch the diagnosis, NBD to drain the infected bile, a fundus first partial cholecystectomy and primary repair of CBD, followed by a peroperative T-tube cholangiogram, usually leads to a satisfactory outcome. PMID- 9752759 TI - Helicobacter pylori infection in duodenal ulcer with gastric outlet obstruction. AB - One hundred and three patients were included in the study. Thirty seven had duodenal ulcer (DU) (Group I), 35 DU with gastric outlet obstruction (GOO) with presence of an active ulcer in the duodenum (Group II). Thirty one had DU with GOO but no active ulcer (Group III). Presence of H. pylori infection was determined by urease test, serology and/or histology. The prevalence of H. pylori in these groups was compared. Levels of Anti-H. pylori IgG antibody titres were also compared. The patients with duodenal ulcer (DU) were significantly younger (38 +/- 2 years) compared to those with established gastric outlet obstruction without ulcer (45 +/- 2 years) (P = 0.02). The prevalence of H. pylori infection in DU (95%), DU with GOO with ulcer (91%) and DU with GOO but no ulcer (90%) was not significantly different (p > 0.05). Anti-H. pylori IgG antibody titre levels were 72 +/- 6 EU/ml in Group III. The titre levels between Group I and Group III were significantly different (P < 0.05). The prevalence of H. pylori infection is high is patients with DU and is unaltered by gastric outlet obstruction. The presence or absence of an active ulcer with gastric outlet obstruction does not affect its association with H. pylori infection. PMID- 9752760 TI - [How can the carbohydrate moiety of influenza virus HA contribute to the initiation of viral infection?]. PMID- 9752761 TI - [Construction of HCV transgenic mice with Cre/loxP switching expression system]. PMID- 9752762 TI - [HIV-1 integration and viral replication]. PMID- 9752763 TI - [Papillomavirus]. PMID- 9752764 TI - [Comparative biology of human herpesviruses 6 and 7]. PMID- 9752765 TI - [Present status and prospect of entomovirus research]. PMID- 9752766 TI - [The baculoviruses]. PMID- 9752768 TI - [Relationship between parasotoid wasps and polydnaviruses]. PMID- 9752767 TI - [Entomopoxviruses]. PMID- 9752769 TI - [Densovirus]. PMID- 9752770 TI - [Cytoplasmic polyhedrosis virus and polyhedrin]. PMID- 9752771 TI - [Genome organization of insect picorna-like viruses]. PMID- 9752772 TI - [The current problems in organizing medical support for the troops]. PMID- 9752773 TI - [The professionalism of the director of a treatment and prevention institution]. PMID- 9752774 TI - [The medical aspects in the psychological training of the troops]. AB - A study of psychological training methods tested on 318 servicemen--armed conflicts participants, including 22 military doctors, veterans of Afghanistan. The results of the research prove that various departures in psychological traits are highly relevant to extreme situations and that a psychological training programme for the medical service personnel is a necessity. The development of the programme should be based on the troops medical maintenance experience in the armed conflicts. PMID- 9752775 TI - [The organization of medical support in garrisons]. PMID- 9752776 TI - [Biologically active food additives]. AB - More than half out of 40 projects for the medical science development by the year of 2000 have been connected with the bio-active edible additives that are called "the food of XXI century", non-pharmacological means for many diseases. Most of these additives--nutricevtics and parapharmacevtics--are intended for the enrichment of food rations for the sick or healthy people. The ecologicaly safest and most effective are combined domestic adaptogens with immuno-modulating and antioxidating action that give anabolic and stimulating effect,--"leveton", "phytoton" and "adapton". The MKTs-229 tablets are residue discharge means. For atherosclerosis and general adiposis they recommend "tsar tablets" and "aiconol (ikhtien)"--on the base of cod-liver oil or "splat" made out of seaweed (algae). All these preparations have been clinically tested and received hygiene certificates from the Institute of Dietology of the Russian Academy of Medical Science. PMID- 9752777 TI - [The use of sorption materials in the combined treatment of suppurative wounds (a review of the literature)]. PMID- 9752778 TI - [Posterior internal correction and fixation of the spine using a rod system]. PMID- 9752779 TI - [The means for optimizing emergency specialized medical care at the N. N. Burdenko Main Military Clinical Hospital]. PMID- 9752780 TI - [Experience in delivering physiotherapeutic care in local armed conflicts and the ways for its improvement]. PMID- 9752781 TI - [Pneumonias in light thoracic trauma]. PMID- 9752782 TI - [The dynamics of the lipid profile under exposure to pelotherapy in cholecystitis patients]. PMID- 9752783 TI - [Transesophageal electrocardiostimulation in the practical work of an outpatient military medical institution]. PMID- 9752784 TI - [A diabetes mellitus school]. PMID- 9752785 TI - [The use of air-plasma flows in military field surgery and disaster medicine]. AB - Plasma currents of high energy are considered to be very promising in the surgical treatment of modern pathology. The scientists of this country have constructed a portable field surgery apparatus in which a heated atmospheric air assumes a form of a high-temperature thin jet used like a surgeon's scalpel. Experiments and clinical tests proved the efficacy of this microplasmotron that can destroy and coagulate body tissue when treating it with plasma. This new method has, besides hemostatic action, some noticeable antimicrobic effect that leads to acceleration of tissue granulation and wound healing. In specialized and skilled medicare this progressive and priority method should be applied for initial surgical wound treatment, secondary hemorrhage arrests and surgical interventions when a high risk of infection spreading is involved. PMID- 9752786 TI - [Experience in organizing antituberculosis measures in the 40th Combined-Arms Army]. PMID- 9752787 TI - [The reconstruction, by computational and instrumental methods, of the doses of irradiation of the population as a consequence of the atmospheric nuclear tests at the Semipalatinsk proving grounds]. PMID- 9752788 TI - [The resolving capacity of the eye and the visual acuity threshold in flight personnel]. PMID- 9752789 TI - [Professor of surgery, military physician A. S. Tauber (on the 150th anniversary of his birth)]. PMID- 9752790 TI - [The 190th anniversary of the Tbilisi Military Hospital]. PMID- 9752791 TI - [The 40th anniversary of the Department of Anesthesiology and Resuscitation of the Military Medical Academy]. PMID- 9752792 TI - [The anniversary of the Department of Military and Emergency Medicine of the Amur State Medical Academy]. PMID- 9752793 TI - Induction of apoptosis in colon cancer cells by nonsteroidal anti-inflammatory drugs. AB - Epidemiological studies have demonstrated that nonsteroidal anti-inflammatory drugs (NSAIDs) decrease the incidence of colon cancer. In addition, NSAIDs reduce the number and size of polyps in patients with familial adenomatous polyposis. The mechanisms of the anti-neoplastic effect of NSAIDs are still far from complete understanding, but one possible mechanism is the induction of apoptosis. Several lines of evidence suggest that NSAIDs-induced apoptosis in colon cancer cells are mediated through the cyclooxygenase (COX)-independent pathway. In this study we explored the mechanism of NSAIDs-induced apoptosis in the colon cancer cell line, HT-29. We confirmed that NSAIDs induce apoptosis in HT-29 cells irrespective of their COX-selectivity. Indomethacin enhanced the expression of p21waf-1 in HT-29 cells. However the expression of apoptosis-related genes such as Fas, bcl-2 and bax was not affected by indomethacin. Intra- and extra-cellular calcium chelators, protein tyrosine kinase (PTK) inhibitor, protein kinase A (PKA) inhibitor and protein kinase C (PKC) inhibitors did not influence indomethacin-induced apoptosis in HT-29 cells. We concluded that NSAIDs-induced apoptosis in colon cancer cells may be independent from signals transducted through [Ca++]i, PTK, PKA, PKC or the expression of apoptosis-related genes. In contrast, our results demonstrating the induction of p21waf-1 transcription by NSAIDs suggest the possible association of NSAIDs-induced apoptosis and cell cycle control in colon cancer cells. PMID- 9752794 TI - The changes in main pulmonary artery pressure after bilateral lung autotransplantation in dogs. AB - In the Respiratory Center, Yongdong Severance Hospital, Yonsei University, we performed 10 cases of bilateral lung autotransplantation in mongrel dogs from July 1994 to June 1996, and we have analyzed the hemodynamic changes. Autotransplantation was performed in order to avoid postoperative rejection. The lung was flushed with an Euro-Collins(E-C) solution containing PGE1 which passed through a 10 French catheter inserted into an incision on the anterior half of the pulmonary artery to pulmonary parenchyme, and the vertical incision was made on the anterior half of the left atrium near the proximal part of the pulmonary vein. However, the bronchus was totally divided after clamping both sides. The lung was kept cold (4 degrees C) in the thoracic cavity for an hour by using slushed ice made from an E-C solution. After an hour of cold ischemia, the pulmonary artery was sutured with Prolene 5-0. The pulmonary vein was sutured with Prolene 6-0 by using the continuous everting mattress method. The main bronchus was anastomosed using the telescope method. The arterial oxygen concentration and the pressures in the femoral and pulmonary arteries were measured both preoperatively and postoperatively. There were no significant hemodynamic differences between the preoperative and postoperative mean pulmonary artery pressure (MPAP) (paired t-test, P = 0.05). Lung preservation using a cold (4 degrees C) E-C solution containing PGE1 may be an acceptable method for short term storage of a lung (for about an hour) in bilateral lung autotransplantation in dogs. PMID- 9752795 TI - Spectral analysis of heart valve sound for detection of prosthetic heart valve diseases. AB - The spectral analysis of heart valve sound is a noninvasive diagnostic method known to be useful in evaluating the state of the heart valve function. This may provide early detection of valve calcification, thrombus or destruction, since previous studies have shown that the dominant frequency peak moved to a high frequency area when natural heart valve leaflets were calcified, stiffened or destroyed. However, it is important for a heart valve sound diagnostic system to find a proper spectral analysis method on phonocardiography. Until now, conventional frequency analyses such as the Fourier transform or autoregressive spectral estimation technique have been used to estimate spectral components of a phonocardiogram, but they are inappropriate because the signal frequency is assumed to remain constant during the transform interval. To overcome this problem, in this study, FOS (Fast Orthogonal Search) & MUSIC (MUltiple SIgnal Classification), which both appeared suitable for the analysis of biological data, were applied to prosthetic heart valve sound as the new heart valve sound spectral analysis methods. Five subjects with normally functioning mechanical heart valves and a patient with a malfunctioning one were selected to collect the heart valve sound signals. As a result, the second dominant peak frequency proved to be important along with the first dominant peak frequency in identifying the valve function. This study showed that the new heart valve sound spectral analysis method presented in this paper may be an effective method in heart valve sound analysis. Further study using this system in a large population of patients will aid in providing a diagnostic method in the early detection of valve failure. PMID- 9752796 TI - Treatment of chronic hepatitis B in children with prednisolone withdrawal followed by recombinant interferon alpha. AB - Steroid withdrawal followed by interferon therapy is an alternative approach for treating chronic hepatitis B virus infection when there has been no therapeutic response to interferon alone. The effectiveness of steroid withdrawal followed by interferon therapy and factors predictive of the response were evaluated in 35 children with biopsy-proven chronic hepatitis B. Patients had received a 1-month course of prednisolone, 1 mg/kg per day orally, followed by a 2-week rest, and then were treated with interferon alpha 3 MU three times per week for 4-6 months. The serum aminotransferase values normalized in 80%, and negative seroconversion rates of HBeAg and HBV-DNA were 69% and 66%. The good response rate was associated with a pretreatment HBV-DNA level lower than 100 pg/ml and a posttreatment ALT level more than 200 IU/L. Normalization of ALT values usually took 5 months, and the clearance of HBV-DNA and HBeAg took 7.8 and 6.7 months, respectively. These results suggest that steroid withdrawal followed by interferon therapy is useful in the treatment of chronic hepatitis B in children, and that a good response rate can be expected in children with lower pretreatment HBV-DNA levels (< 100 pg/ml). PMID- 9752797 TI - Comparison of corneal centering in photorefractive keratectomy. AB - The present study compares three centering methods for excimer laser photorefractive keratectomy (PRK. VISX 20/20) by analyzing the corneal topography. The subjects were grouped according to three different centering methods used in the procedure: an ablation using a light reflex from the patient's cornea pursued by both eyes of the surgeon (Group 1, n = 49); an ablation using a red light reflex from the patient's cornea pursued by the surgeon's left eye only while the right eye remained closed (Group 2, n = 27); an ablation using the patient's center of the pupil pursued by the surgeon's left eye only while the right eye remained closed (Group 3, n = 21). The mean distance from the center of ablation zone to the center of the pupil were; 0.69 +/- 0.45 mm for Group 1, 1.05 +/- 0.48 mm for Group 2 and 0.63 +/- 0.28 mm for Group 3. The degree of deviation in Group 2 was significantly greater than in Group 1 or Group 3. The deviation was greater in the right eyes than the left eyes in Group 2 only. The decentration of the right eye in Group 2 was due to angle Kappa with misalignment of the fixation light and viewing tube containing reticule. PMID- 9752798 TI - Cerebral hemodynamic changes induced by sympathetic stimulation tests. AB - Sympathetic neuronal activity is primarily responsible for the neurogenic control of cerebral autoregulation. The stimulation of sympathetic nerves causes both large arterial constriction and small vessel dilation in experimental animals. However, the role of the sympathetic nervous system in the control of cerebral hemodynamics has yet to be clarified in humans. In order to assess the effect of sympathetic activation on human cerebral hemodynamics, we performed a simultaneous transcranial Doppler (TCD) monitoring of bilateral middle cerebral arterial flow velocity in 16 healthy male volunteers (mean age 26) during well known sympathetic activation measures such as isometric hand-grip exercise (IHE) and cold pressor test (CPT). Blood pressure was checked manually before and at each minute during tests. The mean arterial pressure (MAP) was calculated as (systolic pressure + 2 X diastolic pressure)/3. There was a significant increase in MCA flow velocities during both sympathetic activation tests. The percent increase of diastolic velocity (36% with IHE and 24% with CPT) was significantly higher than systolic velocity (21% with IHE and 9% with CPT). The pulsatility index was significantly decreased during the tests (from 0.75 to 0.58 with IHE and from 0.81 to 0.63 with CPT). These results suggest that sympathetic activation increases MCA flow velocities, related with a reduction in small vessel resistance and/or a constriction of large arteries. PMID- 9752799 TI - Ex vivo generation of functional dendritic cells from mobilized CD34+ hematopoietic stem cells. AB - The ability to generate dendritic cells (DCs) in sizeable numbers has enormous implications for the development of clinically-effective antigen presentation procedures for cancer immunotherapy. We evaluated the generation of immunostimulatory DCs from peripheral blood CD34+ cells collected from healthy donors. CD34+ cells purified from leukapheresis product were seeded at 1 x 10(4) cells/mL in complete medium supplemented with GM-CSF, TNF alpha, IL-4, c-kit ligand, and flt3 ligand (FL). By day 14 of culture in the presence of GM-CSF + TNF alpha, the total cell number increased by 23.4 +/- 5.4-fold compared to the starting number of CD34+ cells. When the c-kit and FL were added to GM-CSF and TNF alpha, the cell number increased by 109.8 +/- 11.2-fold without affecting the immunophenotype of recovered cells. Flow cytometric analysis indicated that cells with the markers of mature dendritic cells, i.e., CD1a +CD14 -HLA-DR+, and CD80+CD86+HLA-DR+, constituted 49.0% +/- 7.5%, and 38.9% +/- 6.5%, respectively. This pattern of expression of surface antigen was unchanged whether the c-kit ligand and/or FL was added. The irradiated CD1a+HLA-DR+ cells recovered from in vitro cultures elicit a vigorous proliferation of allogeneic peripheral blood T cells, irrespective of cytokine combinations. These findings provide advantageous tools for the large-scale generation of DCs that are potentially usable for clinical protocols of immunotherapy or vaccination in patients undergoing cancer treatment. PMID- 9752800 TI - Complement-fixing abilities and IgG subclasses of autoantibodies in epidermolysis bullosa acquisita. AB - Epidermolysis bullosa acquisita (EBA) is an autoimmune-mediated subepidermal bullous disease in which the target of the autoantibodies is type VII collagen, a major component of anchoring fibrils. The purpose of this study was to evaluate the complement-fixing abilities and IgG subclass distribution of autoantibodies in EBA, and to also attempt to investigate the relation between inflammation, complement fixation and IgG subclass distribution in EBA patients. Only 2 sera of 18 patients (11%) showed weak complement-fixing abilities. IgG1 and IgG4 were the most frequently and intensely stained IgG subclasses in EBA sera. We could not find any relationship between the clinico-pathologic types, complement-fixing abilities and IgG subclasses in EBA. These results suggested that complement activation may not be a key factor of bulla formation in EBA. PMID- 9752801 TI - Experimental hypercholesterolemia induces ultrastructural changes in the elastic laminae of rabbit aortic valve. AB - Atherosclerosis is the most severe problem in the high-pressure systemic circulation and similar changes also occur in the high-pressure loading valve. This study was designed to test the hypothesis that early atherosclerosis, induced by a high cholesterol diet in rabbits, is characterized by significant ultrastructural change in the elastic laminae of the aortic valve. However, it is not known whether this process is also taking place in the cardiac valve at the early stage of atherosclerosis. Animals were fed either a high cholesterol diet (n = 5) or a control diet (n = 5) for 10-12 weeks. Histologic analysis demonstrated that subendothelial thickening and foam-cell infiltration were evident in the arterialis of aortic valves. Confocal microscopy revealed an altered pattern characterized by fragmentation and disorganization of the arterialis elastic laminae of hypercholesterolemic valves. Computerized digital analysis of the images obtained by confocal scanning microscopy demonstrated that compared to normal valves, the arterialis elastic laminae of hypercholesterolemic valves decreased in percentage of their elastin content (29.03 +/- 1.10% vs. 42.94 +/- 1.35%, p = 0.023). Immunohistochemical staining for matrix metalloproteinase-3 (MMP-3) revealed MMP-3 immunoreactivity was increased in hypercholesterolemic valves, predominantly in the arterialis. This study demonstrated that early atherosclerosis, induced by a high cholesterol diet in rabbits, is characterized by significant ultrastructural change in the elastic laminae of the aortic valve. The arterialis endothelium of the aortic valve may be a more atherosclerosis-prone area compared with the ventricularis. The presence of ultrastructural defect in the elastic laminae may play a role in chronic degenerative change and a resultant valvular dysfunction. PMID- 9752802 TI - Dietary iodine intake and urinary iodine excretion in normal Korean adults. AB - Korea is a region abundant in foods containing iodine such as seaweed and fish. An adequate amount of iodine consumption is extremely important as both a deficiency and excess of iodine can result in health problems. This study was undertaken to assess the iodine nutritional status of normal Korean adults who consume seaweed and fish, and to determine the relationship between the dietary iodine intake and the urinary excretion of iodine. The dietary assessment of iodine using a food frequency questionnaire and a urinary iodine excretion examination were carried out in 278 healthy adults. The iodide selective electrode (ISE) method was used to determine urinary iodine excretion. The average usual iodine intake of Korean adults was 479 micrograms per day (ranging from 61 micrograms to 4086 micrograms). There was no significant difference in sex or age. The major food sources of dietary iodine included seaweed (66%), milk and dairy products (11%), and fish (9%). The contribution of seaweed to the total iodine intake tended to increase with age while the contribution of milk decreased. The average urinary excretion of iodine was 674 micrograms/g creatinine and there was no significant difference in sex or age. The dietary iodine intake was positively correlated with the urinary excretion of iodine (gamma = 0.60, p < 0.01). The study data indicated that the iodine intake and excretion of Koreans depends mostly on the amount of seaweed consumption like sea tangle and sea mustard. As well, the current iodine intake and urinary iodine excretion by Koreans seems to be higher than in other countries. PMID- 9752803 TI - The effects of diffusion blur on Snellen and grating acuity and foveal function in amblyopia. AB - In order to verify that the effects of diffusion blur on Snellen and grating acuity in amblyopic eyes resemble those obtained from the peripheral or central retina in normal controls, we conducted the following experiment using a liquid crystal window (Edmund Scientific Co.) to produce diffusion blur on Snellen and grating acuity. Spatial frequencies used for a Snellen chart and Teller acuity card were 3.2, 6.5, 13.0, 26.0 cyc/cm at a working distance of 55 cm. The values of diffusive blur on central and peripheral visual acuity obtained from 20 normal healthy control eyes were compared with those values of central visual acuity in 26 amblyopic eyes. The diffusion blur had a strong negative effect on both Snellen and grating acuity in amblyopic eyes, but it had more potent effects on grating acuity (p < 0.05). The diffusion blur values obtained from the central amblyopic retina were more compatible with those of the central retina than with those of the peripheral retina in the control group (p > 0.05). Snellen acuity obtained from diffusion blur overestimated grating acuity in the normal central acuity group and amblyopic central acuity group. The result of this investigation demonstrated that the liquid crystal diffusion blur had a strong negative effect on both Snellen and grating acuity and suggested that the visual function of an amblyopic retina resembled that of a normal central retina. PMID- 9752804 TI - Sclerotherapy with OK-432 for recurrent cystic thyroid nodule. AB - We have adopted OK-432 as a sclerosing agent in the treatment of cystic predominant thyroid nodules and have analyzed our findings to assess the efficacy of intralesional instillation of OK-432. From 1992 through 1993, 48 patients with recurrent or progressive cystic thyroid nodules after 2 or 3 aspirations alone, and whom were cytologically negative for malignancy, were used for this study. The OK-432 solution was prepared by dissolving 0.1 mg of OK-432 in 2 ml of physiologic saline and it was instilled in the amount of 1/10-to-1/5 of the aspirated cystic fluid. A repeated course of therapy was given up to 3 times when sufficient resolution was not obtained 4-to-6 weeks after treatment. The mean number of treatment sessions per patient was 1.5. Throughout the follow-up period from 30-to-45 months (mean, 38 months), 32 (66.7%) patients experienced an almost complete disappearance (< 0.05 cm in diameter) of the cystic lesion, and 12 (25%) patients responded partially by having it decrease by more than half (> 0.5 cm in diameter) of the initial cyst size, and none of these patients required further treatment. The remaining 4 (8.3%) patients showed insufficient resolution despite 3 courses of therapy and 2 of these patients underwent thyroidectomy, in which the lesion proved benign in both cases. All of the patients tolerated the sclerotherapy well. No significant local complications attributed to this treatment were observed. However, a low-grade fever was observed in 26 (54.2%) patients for 2 to 5 days after instillation, which subsided with symptomatic treatment. On the basis of our experience, OK-432 sclerotherapy appears to be safe, simple and effective, and can be a useful alternative treatment for cystic thyroid nodules. PMID- 9752805 TI - Effectiveness of prenatal ultrasonography in detecting fetal anomalies and perinatal outcome of anomalous fetuses. AB - A retrospective study was performed over a 5-year period (1990-94) to evaluate the effectiveness of prenatal ultrasonography in terms of sensitivity, specificity, and predictive values in detecting fetal anomalies by comparing prenatal ultrasonic results with anomalies found in neonates and the perinatal outcome of anomalous fetuses. Minor congenital anomalies as listed and defined in the Eurocat Register were excluded. From a total of 5544 singletons, 4819 had at least one ultrasound scan (87%), of which 3004 at low risk and 1815 (38%) at high risk for anomalies had routine screening (RS) and indicated scanning (IS), respectively. A total of 136 fetuses were structurally abnormal (2.82%, RS and IS: 0.77% and 6.23%) and 200 major anomalies (RS and IS: 37 and 163) were recorded. The overall sensitivity of the ultrasound test was 78.7% (RS and IS: 34.8% and 87.6%, P < 0.01) for abnormal fetuses and 58.0% (RS and IS: 29.7% and 64.4%, P < 0.01) for anomalies. The overall specificity was 99.9% and the positive and negative predictive values were 97.3% and 99.4%, respectively; these values did not differ significantly between the two groups. The sensitivity of ultrasound for the detection of abnormal fetuses before 24 weeks was 22.8% (RS and IS: 13.0% and 24.8%) which was associated with a 61% (25/41) termination rate (RS and IS: 25% and 75.9%, P < 0.01) and a 24.4% (10/41) postnatal survival rate (RS and IS: 41.7% and 17.2%). The overall survival rate following pre- and postnatal correction of anomalies was 44.9% (RS and IS: 60.9% and 41.6%). For the detection of fetal anomalies anatomic ultrasound scanning is necessary during pregnancy, irrespective of pregnancy condition. Early detection of fetal anomalies could offer the option of pregnancy termination. PMID- 9752806 TI - A case of primary bone lymphoma associated with acquired immunodeficiency syndrome. AB - A 33-year old man with acquired immunodeficiency syndrome was admitted to Severance hospital following 1 year of diarrhea and 2 to 3 months of low sternal pain. The patient had progressive generalized lymphadenopathy for the previous 3 years. Whole body bone scan for evaluation of bone pain showed multiple abnormal hot uptakes at the low sternal body and T8 and T10 vertebra. Chest CT showed multifocal cortical erosion of the bone with soft tissue mass at the low sternal body and spine MRI showed multiple low-signal density in T1WI and high-signal density in T2WI at the T8 and T10 vertebral body. Biopsy was performed at the sternochondral junction and it showed high-grade malignant lymphoma of the large cell immunoblastic type. Immunostaining showed positive for the B-cell markers (CD79a and L26) and negative for the T-cell marker (UCHL1). Radiotherapy of 3,000 cGy was delivered to the sternum and vertebra. Since then, systemic chemotherapy with m-BACOD regimen (except dexamethasone) and anti-retroviral therapy with a combination of 3 drugs (didanosine, lamivudine, indinavir) has been performed. This is the first case report of primary bone lymphoma associated with acquired immunodeficiency syndrome in Korea. PMID- 9752807 TI - A case with sarcomatoid hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC) with sarcomatous features is a rare neoplasm which has been found in only 1.8% of surgically resected HCC and has a higher incidence of metastasis than usual HCC. We recently experienced a case of sarcomatoid HCC removed from a 49-year-old man. A surgically resected liver revealed a well defined grayish-white solid firm mass showing extensive central necrosis and infiltrative growth margin. Microscopically, the entire tumor was composed of pleomorphic spindle cells with prominent nucleoli and frequent mitosis. It showed a sinusoidal infiltrative growth pattern at the tumor-nontumor boundary. The tumor cells reacted positively with AE3 (high molecular cytokeratin) and Vimentin and reacted negatively with AE1 (low molecular cytokeratin), cytokeratin19, carcinoembryonic antigen, alpha-fetoprotein, Factor VIII, CD31 and CD68. The spindle-shaped tumor cells were considered to originate from hepatocyte rather than from bile duct epithelium or mesenchymal elements. PMID- 9752808 TI - Facing up to responsibilities in science. PMID- 9752809 TI - Is it time for a new injury score? PMID- 9752810 TI - Interpretation of blood lactate concentrations in patients with sepsis. PMID- 9752811 TI - Melanoma and the radiograph: sanctity of human life. PMID- 9752812 TI - Multifactorial approach to stroke investigation and prevention. PMID- 9752813 TI - Malaria disaster in Africa. PMID- 9752814 TI - Randomised trial of effectiveness of second eye cataract surgery. AB - BACKGROUND: The effectiveness of cataract surgery on one eye is well established, but concerns over health-care expenditure have called into question the value of cataract surgery on the second eye. We examined the effects of second eye surgery in terms of patient perceptions as well as through visual acuity, contrast sensitivity, and stereoacuity tests. METHODS: 208 otherwise healthy patients awaiting second eye cataract surgery were recruited into our randomised trial. At randomisation participants were allocated expedited surgery (planned to take place within 6 weeks) or routine surgery (routine waiting time, 7-12 months). Assessments were made at randomisation and again at review after about 6 months. Eight primary trial outcomes consisted of four questionnaire items and four visual function tests, done with both eyes open. FINDINGS: Traditional clinical tests of success in cataract surgery (visual acuity and contrast sensitivity) showed only slight differences in binocular vision in favour of the expedited surgery group. There were major benefits for the expedited-surgery group in terms of reported visual symptoms and effects on quality of life. At review, differences in self-reported vision related difficulties between the two groups ranged from 11% (95% CI 4.4-17%, activities) to 30% (19-41%, reading). Stereoacuity was better in the expedited surgery group, the difference between the groups for the proportions with stereoacuity of 3000 s of arc or worse was 58% (47-69%). INTERPRETATION: This trial has shown that there is a clear benefit from second eye cataract surgery. PMID- 9752815 TI - Polychemotherapy for early breast cancer: an overview of the randomised trials. Early Breast Cancer Trialists' Collaborative Group. AB - BACKGROUND: There have been many randomised trials of adjuvant prolonged polychemotherapy among women with early breast cancer, and an updated overview of their results is presented. METHODS: In 1995, information was sought on each woman in any randomised trial that began before 1990 and involved treatment groups that differed only with respect to the chemotherapy regimens that were being compared. Analyses involved about 18,000 women in 47 trials of prolonged polychemotherapy versus no chemotherapy, about 6000 in 11 trials of longer versus shorter polychemotherapy, and about 6000 in 11 trials of anthracycline-containing regimens versus CMF (cyclophosphamide, methotrexate, and fluorouracil). FINDINGS: For recurrence, polychemotherapy produced substantial and highly significant proportional reductions both among women aged under 50 at randomisation (35% [SD 4] reduction; 2p<0.00001) and among those aged 50-69 (20% [SD 3] reduction; 2p<0.00001); few women aged 70 or over had been studied. For mortality, the reductions were also significant both among women aged under 50 (27% [SD 5] reduction; 2p<0.00001) and among those aged 50-69 (11% [SD 3] reduction; 2p=0.0001). The recurrence reductions emerged chiefly during the first 5 years of follow-up, whereas the difference in survival grew throughout the first 10 years. After standardisation for age and time since randomisation, the proportional reductions in risk were similar for women with node-negative and node-positive disease. Applying the proportional mortality reduction observed in all women aged under 50 at randomisation would typically change a 10-year survival of 71% for those with node-negative disease to 78% (an absolute benefit of 7%), and of 42% for those with node-positive disease to 53% (an absolute benefit of 11%). The smaller proportional mortality reduction observed in all women aged 50-69 at randomisation would translate into smaller absolute benefits, changing a 10-year survival of 67% for those with node-negative disease to 69% (an absolute gain of 2%) and of 46% for those with node-positive disease to 49% (an absolute gain of 3%). The age-specific benefits of polychemotherapy appeared to be largely irrespective of menopausal status at presentation, oestrogen receptor status of the primary tumour, and of whether adjuvant tamoxifen had been given. In terms of other outcomes, there was a reduction of about one-fifth (2p=0.05) in contralateral breast cancer, which has already been included in the analyses of recurrence, and no apparent adverse effect on deaths from causes other than breast cancer (death rate ratio 0.89 [SD 0.09]). The directly randomised comparisons of longer versus shorter durations of polychemotherapy did not indicate any survival advantage with the use of more than about 3-6 months of polychemotherapy. By contrast, directly randomised comparisons did suggest that, compared with CMF alone, the anthracycline-containing regimens studied produced somewhat greater effects on recurrence (2p=0.006) and mortality (69% vs 72% 5 year survival; log-rank 2p=0.02). But this comparison is one of many that could have been selected for emphasis, the 99% CI reaches zero, and the results of several of the relevant trials are not yet available. INTERPRETATION: Some months of adjuvant polychemotherapy (eg, with CMF or an anthracycline-containing regimen) typically produces an absolute improvement of about 7-11% in 10-year survival for women aged under 50 at presentation with early breast cancer, and of about 2-3% for those aged 50-69 (unless their prognosis is likely to be extremely good even without such treatment). Treatment decisions involve consideration not only of improvements in cancer recurrence and survival but also of adverse side effects of treatment, and this report makes no recommendations as to who should or should not be treated. PMID- 9752816 TI - Influences of educational interventions and adverse news about calcium-channel blockers on first-line prescribing of antihypertensive drugs to elderly people in British Columbia. AB - BACKGROUND: The way in which dissemination of evidence changes medical practice needs to be better understood. Controversy about calcium-channel blockers (CCB) in the past 3 years has provided a natural experiment, enabling assessment of the impact of media stories, a national warning letter, a teleconference, small group workshops, and newsletters on first-line prescribing of antihypertensive drugs. METHODS: We included all physicians (4403) in British Columbia who prescribed a thiazide diuretic, beta-blocker, inhibitor of angiotensin-converting enzyme (ACE), or CCB as the first antihypertensive agent for 36,507 residents aged 66 years and over, with no previous or concurrent sign of underlying cardiovascular disease. We used a database covering all prescriptions to elderly people to measure the change in proportion of newly treated patients who received each class of drug as first-line therapy. We used a matched cohort design for assessment of the teleconference and workshops, a randomised community design for the newsletters, and time-series analysis for the media impacts. FINDINGS: The proportion of patients who received a CCB as first-line therapy declined gradually from 22% in early 1994 to 15% in late 1996. This proportion was not affected by two waves of adverse news about CCBs in 1995, but fell by 5% for 5 months and by 3% for 1 month after two waves in 1996. The proportion of patients who received either a CCB or an ACE inhibitor as first-line therapy, contrary to guidelines, was still 42% overall in 1996. The workshops and newsletters were followed by shifts from first-line CCB to first-line thiazide prescribing. INTERPRETATION: Changes in prescribing practices occur gradually with the accumulation of small impacts from educational interventions and lay media attention. PMID- 9752817 TI - Bone scintigraphy in screening of torture survivors. AB - BACKGROUND: In most developed countries, survivors of physical torture inflicted for political, religious, or ethnic reasons face ever more stringent review when seeking asylum. In Austria, asylum seekers are required by immigration authorities to undergo medical examination as part of the review. Bone scintigraphy can detect bone lesions that are not detectable clinically or radiologically. We assessed the value of bone scintigraphy as corroboration of alleged injuries. METHODS: Human-rights organisations referred 25 asylum seekers to us from countries where torture is practised. We included patients who claimed to have been beaten by the security forces in their home country because of political or religious conviction or ethnic origin. Injuries had been inflicted 4 months to 5 years earlier. The patients (three women, 22 men) from 12 countries were categorised retrospectively into two groups: group A (n=12), tortured with blows from hard objects, and group B (n=13), tortured with blows from fists and kicks. We also used a control group of 25 individuals with the same age and sex distribution from the same countries who had no history of torture. FINDINGS: In group A, bone scans showed abnormalities in the area of alleged injury in all patients, whereas radiography was positive in only five patients. In group B, bone scans in the alleged areas of damage were positive in seven patients, but radiography yielded no positive outcomes. Among the controls there was one abnormal scan due to a known coxarthrosis. INTERPRETATION: Our preliminary results suggest that bone scintigraphy is a sensitive, non-invasive tool to document trauma some years after the actual injury. PMID- 9752818 TI - Focal cortical release of endogenous opioids during reading-induced seizures. AB - BACKGROUND: Studies in animals implicate endogenous release of opioid peptides as a mechanism for terminating partial and generalised seizures. To localise dynamic changes in opioid neurotransmission associated with partial seizures and higher cognitive function, we investigated the release of endogenous opioids in patients with reading-induced seizures compared with healthy controls. METHODS: Five patients who had reading epilepsy and six controls had 11C-diprenorphine (DPN) positron-emission-tomography (PET) scans while reading a string of symbols (baseline) or a scientific paper (activation). Statistical parametric mapping was used to find areas with differences in opioid-receptor binding. FINDINGS: On activation scans mean 11C-DPN binding to opioid receptors was significantly lower (p<0.05 corrected for multiple non-independent comparisons) in the left parieto temporo-occipital cortex (Brodmann area 37) in reading-epilepsy patients compared with controls. INTERPRETATION: These findings suggest that opioid-like substances are involved in the termination of reading-induced seizures. PMID- 9752819 TI - A man with diabetes and unexplained renal failure. PMID- 9752820 TI - Acute myocardial infarction associated with sildenafil. PMID- 9752821 TI - Tolcapone and fulminant hepatitis. PMID- 9752822 TI - Ciliary neurotropic factor in skin biopsies of patients with amyotrophic lateral sclerosis. PMID- 9752823 TI - Association of mannose-binding-lectin deficiency with severe atherosclerosis. PMID- 9752824 TI - Oxidative stress after thrombolysis. PMID- 9752825 TI - Mosaic atopic eczema cured by autotransplantation? PMID- 9752826 TI - Normal telomere lengths in young mothers of children with Down's syndrome. PMID- 9752827 TI - Homemade anticoagulation monitor. PMID- 9752828 TI - Treatment options clearer for prostate cancer. PMID- 9752829 TI - Current assays for HHV-8 seropositivity unreliable. PMID- 9752830 TI - Treating type 1 diabetes by regulating autoimmunity. PMID- 9752831 TI - NIH's big budget gains face new uncertainty. PMID- 9752832 TI - South Africa holds crisis summit on HIV. PMID- 9752833 TI - "The flying hospital" visits UK airshow. PMID- 9752834 TI - Dengue and dengue haemorrhagic fever. AB - The incidence and geographical distribution of dengue have greatly increased in recent years. Dengue is an acute mosquito-transmitted viral disease characterised by fever, headache, muscle and joint pains, rash, nausea, and vomiting. Some infections result in dengue haemorrhagic fever (DHF), a syndrome that in its most severe form can threaten the patient's life, primarily through increased vascular permeability and shock. The case fatality rate in patients with dengue shock syndrome can be as high as 44%. For decades, two distinct hypotheses to explain the mechanism of DHF have been debated-secondary infection or viral virulence. However, a combination of both now seems to be the plausible explanation. The geographical expansion of DHF presents the need for well-documented clinical, epidemiological, and virological descriptions of the syndrome in the Americas. Biological and social research are essential to develop effective mosquito control, medications to reduce capillary leakage, and a safe tetravalent vaccine. PMID- 9752835 TI - Heading up the American dream of health. Interview by David H Frankel and Julie Rovner. PMID- 9752836 TI - Caution: should we be treating HIV infection early? PMID- 9752837 TI - DeltaF508 heterozygosity and asthma. PMID- 9752838 TI - DeltaF508 heterozygosity and asthma. PMID- 9752839 TI - DeltaF508 heterozygosity and asthma. PMID- 9752840 TI - DeltaF508 heterozygosity and asthma. EGEA Co-operative Group. PMID- 9752841 TI - DeltaF508 heterozygosity and asthma. PMID- 9752842 TI - Dietary sodium intake and mortality: NHANES. The Faculty 31st International Society and Federation of Cardiology 10-day Teaching Seminar in Cardiovascular Disease, Epidemiology and Prevention. National Health and Nutrition Examination Survey. PMID- 9752843 TI - Prognostic value of Th1/Th2 ratio in rheumatoid arthritis. PMID- 9752844 TI - Prenatal DNA diagnosis of Down's syndrome by PCR. PMID- 9752845 TI - Prenatal DNA diagnosis of Down's syndrome by PCR. PMID- 9752846 TI - Mandatory hepatitis B virus testing for doctors. PMID- 9752847 TI - Surveillance into crowd control agents. PMID- 9752848 TI - Placebo needle for acupuncture. PMID- 9752849 TI - Nursing nursing back to health. PMID- 9752850 TI - High frequency radio keeps mosquitoes at bay. PMID- 9752851 TI - Sketches from The Lancet. PMID- 9752852 TI - Science's own superstition. PMID- 9752853 TI - Audit of the surgeon: problems and perspectives. PMID- 9752854 TI - Cryoablation of liver tumours. PMID- 9752855 TI - Antireflux surgery in the laparoscopic era. AB - BACKGROUND: The recent development of laparoscopic techniques for fundoplication has created renewed interest in surgery for gastro-oesophageal reflux disease, leading to reports of large clinical series from many centres. However, controversy remains about technical aspects of laparoscopic antireflux surgery, with no consensus yet reached about a standard operative technique. It is important, therefore, to reassess critically the results of laparoscopic surgery for reflux disease, so that its current status can be determined. METHODS: Published outcome studies for laparoscopic antireflux surgery, as well as selected studies from the era of open antireflux surgery, were reviewed to assess outcomes. RESULTS: The results of case series for laparoscopic antireflux surgery with short- and medium-term follow-up, as well as the early results of randomized trials, confirm that this approach reduces the early overall morbidity of surgery for reflux disease. However, certain complications may be more common, for instance paraoesophageal hiatus herniation, pneumothorax and oesophageal perforation, requiring surgeons to use specific strategies which can help to avoid these problems. Published studies and trials do not support the routine or selective application of a posterior partial fundoplication technique or routine division of the short gastric vessels during Nissen fundoplication. CONCLUSION: At present, a short loose Nissen fundoplication performed laparoscopically, with or without division of the short gastric vessels, is an appropriate surgical approach for gastro-oesophageal reflux disease. However, long-term outcomes following laparoscopic antireflux surgery will not be available for some years, and must be awaited before the final status of the various laparoscopic techniques can be confirmed. PMID- 9752856 TI - Co-existing abdominal aortic aneurysm and intra-abdominal malignancy: reflections on the order of treatment. AB - BACKGROUND: The management of simultaneously occurring abdominal aortic aneurysm and intra-abdominal malignancy is controversial. It is unclear whether to treat the aneurysm first or the malignancy, or both simultaneously. If the malignancy is resected first there is a risk of postoperative rupture of the aneurysm. If simultaneous surgery is performed there is a risk of prosthetic graft infection from contamination by gastrointestinal or urinary tract contents. METHODS: Relevant papers from 1960 to 1996, identified from Medline and manual searching, were reviewed. RESULTS AND CONCLUSION: The literature supports the conclusion that the lesion of greater priority is that posing the greater threat to the patient; this is usually the aneurysm, especially if it is over 6 cm in diameter. For renal malignancies simultaneous surgery is the treatment of choice, but for bladder cancer the best management is unclear. Large aneurysms should usually be resected in preference to colorectal cancer unless the cancer is locally advanced, perforated or likely to result in early intestinal obstruction. If both lesions are complicated there may be a case for simultaneous treatment. PMID- 9752857 TI - Chlamydia pneumoniae and vascular disease. AB - BACKGROUND: There is an increasing body of evidence linking the human pathogen Chlamydia pneumoniae with atherosclerosis. METHODS: A Medline-based review of the literature was carried out. RESULTS AND CONCLUSION: Seroepidemiological studies have revealed the possibility that evidence of infection with C. pneumoniae and atherosclerotic disease are related. Studies on human tissue have demonstrated that evidence of the organism can be found in human atherosclerotic tissue by both direct and indirect methods significantly more often than in control vascular tissue. Using animal models it is possible to show that C. pneumoniae can be disseminated haematogenously following pulmonary infection and that it shows a tropism for atherosclerotic tissue. In vitro work has demonstrated that the organism is capable of infecting, surviving and multiplying in cells of the human vascular wall, and that it can provoke a cell-mediated cytokine response which has implications both locally and systemically. Two clinical trials of macrolide antibiotics have demonstrated that they confer increased cardiovascular protection in patients following myocardial infarction. Adequately powered trials are needed to establish the therapeutic role of antibiotics in peripheral arterial disease. PMID- 9752858 TI - Prolonged survival in selected patients following surgical resection for pulmonary metastasis from hepatocellular carcinoma. AB - BACKGROUND: Pulmonary metastasis is the commonest site of extrahepatic spread from hepatocellular carcinoma (HCC). The aim of the present study was to evaluate the efficacy of surgical management in patients with solitary pulmonary metastases from HCC. METHODS: This was a retrospective study of patients with HCC admitted for hepatectomy from July 1972 to June 1995. The records of patients who had a pulmonary resection for histologically proven pulmonary recurrence after curative hepatectomy were selected for analysis. RESULTS: In the study interval, 380 patients with HCC underwent hepatectomy. Some 48 patients (12.6 per cent) developed pulmonary metastases documented pathologically or radiologically. Nine (seven men and two women) were suitable for curative pulmonary resection. The median disease-free survival between hepatectomy and appearance of the lung metastasis was 21 months. The median survival after pulmonary resection was 42 months, and the 1-, 2- and 5-year survival rates were 100, 78 and 67 per cent respectively. CONCLUSION: Pulmonary resection for metastases from HCC resulted in long-term survival in these highly selected patients. PMID- 9752859 TI - Effect of lithium gamma-linolenate on the growth of experimental human pancreatic carcinoma. AB - BACKGROUND: The lithium salt of gamma-linolenic acid (Li-GLA) is growth inhibitory to pancreatic cancer cells in vitro and is reported to prolong the survival of patients with pancreatic cancer. The effect of Li-GLA on the growth of human pancreatic carcinoma in vivo is not known. In this study the effect of parenterally administered Li-GLA on the growth of human pancreatic carcinoma in nude mice was tested. METHODS: Pancreatic tumours were produced in nude mice by subcutaneous implantation of MIA PaCa-2 cells. This cell line is sensitive to Li GLA in vitro. Mice were randomly treated with intraperitoneal, intravenous or intratumoral Li-GLA. Each group also had controls. RESULTS: Both intravenous and intraperitoneal administration of Li-GLA had no significant effect on tumour growth or tumour phospholipid fatty acid composition. Intratumoral administration of Li-GLA was, however, associated with a significant antitumour effect. CONCLUSION: Within the limitations of this tumour model, the benefit seen with intravenous Li-GLA in patients with pancreatic carcinoma cannot be explained by tumour growth inhibition. Local administration appears to be more effective than intravenous or intraperitoneal therapy. PMID- 9752860 TI - Intrahepatic peripheral cholangiocarcinoma: mode of spread and choice of surgical treatment. AB - BACKGROUND: Classification of macroscopic appearance and standard operative procedures for intrahepatic cholangiocarcinoma (ICC) are still controversial. METHODS: The mode of spread of 12 resected ICCs was examined by light microscopy, and the appropriate operative procedures for the various tumours were considered. RESULTS: Macroscopically, nine tumours were classified as mass-forming type and three as periductal infiltrating type. All patients were treated by major hepatectomy; resection of the extrahepatic bile duct was included in two cases of the periductal infiltrating type. Microscopically, invasion into the portal vein, intrahepatic metastasis and perineural or lymphatic vessel invasion occurred in none, one and all of three tumours of the periductal infiltrating type and in eight, six and six of nine tumours of the mass-forming type. CONCLUSION: ICC of the periductal infiltrating type has a tendency to spread along Glisson's sheath via lymphatic vessels. By contrast, ICC of the mass-forming type tends to invade the liver via the portal vein system; such tumours begin to invade Glisson's sheath through the lymphatic vessels when the tumour has increased in size. Therefore, major hepatectomy with combined resection of the extrahepatic bile duct should be performed for all ICCs of the periductal infiltrating type and for those of the mass-forming type with invasion of Glisson's sheath. PMID- 9752861 TI - Preliminary experience with percutaneous cryotherapy of liver tumours. PMID- 9752862 TI - Experimental study of electrolysis-induced hepatic necrosis. AB - BACKGROUND: One of the most promising but unexplored methods for treating patients with irresectable liver tumours is electrolysis. This study examined the effect of increasing 'current dose' on the volume of the lesion induced in normal rat liver. METHODS: A direct current generator, connected to platinum electrodes implanted in the rat liver, was used to examine the effect of (1) varying current doses from 1 to 5 coulombs and (2) electrode separation (2 or 20 mm), on the volume of liver necrosis. RESULTS: There was a significant correlation (P < 0.001) between the current dose and the volume of necrosis produced for each electrode separation. Placing the electrodes 2 mm apart resulted in smaller total volumes of necrosis than placing them 20 mm apart when anode lesions were significantly larger than cathode lesions (P< 0.05). Liver enzymes (aspartate aminotransferase, alanine aminotransferase) were significantly raised 1 day after treatment (P < 0.001) and predicted the total volume of hepatic necrosis (P < 0.001). CONCLUSION: Predictable and reproducible areas of liver necrosis are produced with electrolysis. If these results extrapolate to larger animal models, this technique has potential for patients with irresectable primary and secondary liver tumours. PMID- 9752863 TI - POSSUM and Portsmouth POSSUM for predicting mortality. Physiological and Operative Severity Score for the enUmeration of Mortality and morbidity. AB - BACKGROUND: There is a need for an accurate measure of surgical outcomes so that hospitals and surgeons can be compared properly regardless of case mix. POSSUM (Physiological and Operative Severity Score for the enUmeration of Mortality and morbidity) uses a physiological score and an operative severity score to calculate risks of mortality and morbidity. In a previous small study it was found that Portsmouth POSSUM (P-POSSUM; a modification of the POSSUM system) provided a more accurate prediction of mortality. METHODS: Some 10000 general surgical interventions (excluding paediatric and day cases) were studied prospectively between August 1993 and November 1995. The POSSUM mortality equation was applied to the full 10000 surgical episodes. The 10000 patients were arranged in chronological order and the first 2500 were used as a training set to produce the modified P-POSSUM predictor equation. This was then applied prospectively to the remaining 7500 patients arranged chronologically in five groups of 1500. RESULTS: The original POSSUM logistic regression equation for mortality overpredicts the overall risk of death by more than twofold and the risk of death for patients at lowest risk (5 per cent or less) by more than sevenfold. The P-POSSUM equation produced a very close fit with the observed in hospital mortality. CONCLUSION: P-POSSUM provides an accurate method for comparative surgical audit. PMID- 9752864 TI - D-lactate as an early marker of intestinal ischaemia after ruptured abdominal aortic aneurysm repair. AB - BACKGROUND: Patients with a ruptured abdominal aortic aneurysm (AAA) are at risk of developing colonic ischaemia after surgery. It is difficult to diagnose this ischaemia at an early stage. D-lactate is produced by intestinal bacteria after ischaemia. L-lactate is released in increased amounts during hypoxia by anaerobic metabolism. This study investigated both variables as a marker for intestinal ischaemia in patients with a ruptured AAA. METHODS: Twenty-four patients with ruptured AAA were divided retrospectively into two groups with and without ischaemic complications, as verified by colonoscopy. Blood had been taken on admission to the intensive care unit (ICU). Median time to colonoscopy was 9 days after surgery. As controls, four patients with pneumonia, six healthy subjects, five patients with an elective AAA repair, and six patients with sepsis and acute tubular necrosis were included. RESULTS: D-lactate level on admission was significantly increased in patients with colonic ischaemia after ruptured AAA compared with the level in patients without ischaemia (P< 0.05), patients with sepsis (P< 0.001), those with pneumonia and healthy subjects (P< 0.01). L-lactate concentration was similar in the group with intestinal complications and in patients without colonic ischaemia; however, L-lactate levels were higher in patients with pneumonia and sepsis than in healthy subjects (P < 0.05). CONCLUSION: On admission to the ICU, D-lactate, but not L-lactate, levels may predict later colonic ischaemia following repair of a ruptured AAA. PMID- 9752865 TI - Predischarge duplex imaging of infrainguinal vein grafts does not predict the development of stenoses. AB - BACKGROUND: The aim of this study was to determine whether predischarge duplex imaging of an infrainguinal vein graft could predict the subsequent development of graft stenosis. METHODS: Patients with infrainguinal vein grafts underwent a duplex scan of the entire graft before discharge from hospital. Grafts were classified as abnormal or normal based on the presence or absence of flow abnormalities (peak velocity ratio 1.5 or greater). All grafts were re-examined 1 month after surgery, at 3-monthly intervals for the first year and then every 6 months thereafter. A significant stenosis requiring correction was defined by a duplex-derived peak systolic velocity ratio of 3.0 or more. The relationship between the predischarge scan and subsequent development of stenosis was examined. RESULTS: Forty-four grafts were recruited. Two occluded soon after a normal predischarge scan and were excluded from subsequent analysis. Predischarge abnormalities were found in 16 grafts. The abnormality in one graft required immediate correction. A further four grafts subsequently developed significant stenoses and required angioplasty. Of the 26 grafts with no predischarge abnormalities, 11 developed significant stenoses and underwent angioplasty. Abnormal predischarge duplex scans had a sensitivity and specificity of 31 and 58 per cent respectively for the development of stenoses. The positive and negative predictive values were 31 and 58 per cent respectively. CONCLUSION: Predischarge duplex imaging cannot be used to predict the development of stenoses in infrainguinal vein grafts. PMID- 9752866 TI - In vivo pressure profiles of thigh-length graduated compression stockings. AB - BACKGROUND: The aim of the study was to assess whether the appropriate pressure profile is generated by thigh-length graduated compression stockings in human subjects. The effect of leg posture on the pressure profile was assessed in three commonly used brands of graduated compression stockings. METHODS: The study involved 17 human volunteers from the Department of Orthopaedic Surgery. Three different brands of stockings commonly recommended for the prevention of deep vein thrombosis were applied to each individual and the interface pressure profile under the stocking was measured. The effect of posture was assessed by comparing the interface pressure profile with the subject supine and in standing and sitting positions. RESULTS: Appropriate median pressure profiles were achieved only with the subject standing or supine. In the sitting position with the knee flexed, a high median interface pressure in excess of 28 mmHg was generated at the popliteal fossa. Overall, inconsistent performance was found in all three brands of stockings; fewer than 30 per cent of the pressure readings fell within 20 per cent of the 'ideal'. Reversed pressure profile was observed in over 70 per cent of subjects. CONCLUSION: Thigh-length graduated compression stockings may be most effective in bedridden patients. Knee-length stockings may be more suitable for the prevention of deep vein thrombosis in ambulant patients. PMID- 9752867 TI - Antimicrobial prophylaxis in colorectal surgery: a systematic review of randomized controlled trials. AB - BACKGROUND: A systematic review was carried out to assess the relative efficacy of antimicrobial prophylaxis for the prevention of postoperative wound infection in patients undergoing colorectal surgery. METHODS: MEDLINE, EMBASE, the Cochrane Trials Register and the references cited in retrieved studies were searched to identify relevant trials published between 1984 and 1995. RESULTS: Some 147 relevant trials were identified. The quality of trials has improved over the past 12 years. The results confirm that the use of antimicrobial prophylaxis is effective for the prevention of surgical wound infection after colorectal surgery. There was no significant difference in the rate of surgical wound infections between many different regimens. However, certain regimens appear to be inadequate (e.g. metronidazole alone, doxycycline alone, piperacillin alone, oral neomycin plus erythromycin on the day before operation). A single dose administered immediately before the operation (or short-term use) is as effective as long-term postoperative antimicrobial prophylaxis (odds ratio 1.17 (95 per cent confidence interval (c.i.) 0.90-1.53)). There is no convincing evidence to suggest that the new-generation cephalosporins are more effective than first generation cephalosporins (odds ratio 1.07 (95 per cent c.i. 0.54-2.12)). CONCLUSION: Antibiotics selected for prophylaxis in colorectal surgery should be active against both aerobic and anaerobic bacteria. Administration should be timed to make sure that the tissue concentration of antibiotics around the wound area is sufficiently high when bacterial contamination occurs. Guidelines should be developed locally in order to achieve a more cost-effective use of antimicrobial prophylaxis in colorectal surgery. PMID- 9752868 TI - Small bowel obstruction after total or subtotal colectomy: a 10-year retrospective review. AB - BACKGROUND: The aim of this retrospective study was to determine the cumulative incidence of adhesive small bowel obstruction (SBO) after total or subtotal colectomy and to investigate the site of the obstructive adhesions in the abdominal cavity. METHODS: The records of 234 patients who underwent colectomy from 1985 to 1994 were reviewed for SBO, potential risk factors for SBO, and the site of adhesions causing obstruction. Mean follow-up, which was complete in 215 patients, was 63 months. RESULTS: SBO occurred in 56 patients (24 per cent) of whom 42 (18 per cent) had adhesive obstruction. The risk of SBO due to adhesions within 1 year was 11 per cent, increasing to 30 per cent 10 years after colectomy. With univariate analysis no risk factor for adhesive SBO, including previous laparotomies, septic complications and omental resection, was identified. The most common site of obstructing adhesions was the pelvis (ten of 28 patients). CONCLUSION: The incidence of SBO after colectomy is high. Colectomy may be a suitable model for studies of adhesion prevention. PMID- 9752869 TI - Long-term clinical outcome of surgery for solitary rectal ulcer syndrome. AB - BACKGROUND: When severe symptoms of solitary rectal ulcer syndrome persist despite medical management, surgery may be necessary. METHODS: A retrospective review was carried out of 81 patients undergoing surgery for solitary rectal ulcer syndrome in the 10-year period from 1984 to 1993 to determine the long-term outcome at a minimum follow-up of 12 months. Of the 81 patients, 15 were excluded from further analysis (11 were followed up for less than 12 months, two died and two were lost to follow-up). Sixty-six patients were studied (median age 38 (range 15-77) years; 53 female). Of these, 49 underwent rectopexy, nine Delorme's operation, two anterior resection and four creation of a stoma as the initial operation. RESULTS: At a median follow-up of 90 (range 12-177) months, the rectopexy had failed in 22 of 49 patients; 19 of these patients underwent further surgery, including rectal resection with coloanal anastomosis (four with three failures), colostomy (11) and other procedures (four). Ultimately, 14 required a stoma. Constipation was the indication for a stoma in nine of the 11 patients who had colostomy as the first procedure after failure of rectopexy. Nine patients had Delorme's operation as the first procedure. At median follow-up of 38 (range 19-107) months, there were four failures. Two of these ultimately required a stoma. Of the seven patients who underwent anterior resection as the initial or subsequent procedure, a stoma was finally necessary in four. Anterior resection used as a salvage procedure was not successful. The overall stoma rate was 30 per cent (20 patients). Of 11 symptoms assessed before operation only incontinence and incomplete evacuation were related to a poor outcome following surgery. CONCLUSION: Antiprolapse operations result in a satisfactory long-term outcome in about 55-60 per cent of patients having surgery for solitary rectal ulcer syndrome. Results of anterior resection are disappointing. PMID- 9752870 TI - Quality of life in patients undergoing treatment for chronic radiation-induced rectal bleeding. AB - BACKGROUND: Laser and formalin therapy have recently been shown to be safe methods of haemostasis in patients with chronic radiation-induced rectal bleeding (CRRB). The effectiveness of this treatment in terms of improved quality of life remains to be determined. The aim of this study was to develop a questionnaire measure of self-perceived quality of life in patients with CRRB. METHODS: A self completed Rectal Bleeding Quality of Life Scale (RBQOLS) based on the social and emotional problems experienced by patients with CRRB was developed using standard psychometric methods. Before laser and formalin therapy, 34 consecutive patients completed the questionnaire which was repeated 1 month after treatment and again 5 months later. The validity of the scale was assessed in relation to patient response to treatment. RESULTS: The RBQOLS had high reliability (alpha=0.89) and its concurrent validity was confirmed by a significant association with the pretreatment severity of CRRB assessed endoscopically. The mean RBQOLS score increased from 102 (95 per cent confidence interval 90-114) before treatment to 126 (111-141) after treatment (t=3.1, 33 d.f., P=0.004) and 136 (122-151) at follow-up attesting to its predictive validity. CONCLUSION: The RBQOLS is a reliable and valid device for assessing quality of life of patients with this uncommon, previously intractable and potentially life threatening complication of radiotherapy. PMID- 9752871 TI - Young age is not a poor prognostic marker in colorectal cancer. AB - BACKGROUND: There is still considerable controversy and debate regarding the features and prognosis of colorectal cancer in young patients. METHODS: One hundred and ten patients (5.1 per cent) under the age of 40 years with colorectal cancer (group Y; male: female ratio 48:62) were compared with 2064 patients with colorectal cancer aged 40 years or more (group O; 917 women, 1147 men). Mode of presentation, stage at diagnosis, tumour characteristics and survival were analysed. RESULTS: Predisposing malignant conditions and family history of colorectal cancer were present in 20.9 per cent of patients in group Y versus 2.2 per cent in group O (P < 0.001). Common chief complaints included change in bowel habits, bleeding from the rectum and a significantly higher incidence of abdominal pain in group Y. There was no difference in stage at presentation between the two groups (the proportion of Dukes stage A, B, C and 'D' lesions in group Y was 8.2, 24.5, 37.3 and 30.0 per cent respectively versus 10.5, 27.9, 33.4 and 28.1 per cent in group O). Tumour site and characteristics were similar in both groups. The incidence of mucinous/signet ring cell and poor grade tumours was 6.9 and 11.8 per cent respectively in group Y and 4.5 and 10.5 per cent in group O. With a mean follow-up of 31.8 months, the overall 5-year survival rate was 54.8 per cent in group Y and 54.1 per cent in group O. Comparing stage for stage, survival was not significantly different in the two groups. However, the adjusted hazard ratios of the age groups Y, M (40-59 years), S (60-79 years), and E (80 years and above) were 1.3, 1 (baseline for calculations), 1.4 and 2.4 respectively, suggesting an adverse outcome for patients in group Y compared with patients aged 40-59 years. CONCLUSION: This study revealed no difference in tumour characteristics and survival in patients with colorectal cancer aged less than 40 years compared with those aged above 40 years. However, a higher hazard ratio in the youngest group may connote a worse prognosis than that for those aged 40-59 years. A significant family history of colorectal cancer and predisposing conditions in the young warrants aggressive screening, surveillance and treatment of the underlying conditions. The detection of colorectal cancer in young patients should be no different from that in the old but demands a high index of suspicion. PMID- 9752872 TI - Improved outcome after emergency surgery for cancer of the large intestine. AB - BACKGROUND: Emergency surgery for colorectal cancer has become more aggressive and radical over the past decade. This retrospective review analyses the impact on outcome. METHODS: The results of emergency surgery within 24 h of admission were compared between 1982 and 1987 (77 patients) and 1988 and 1993 (75 patients). Patient and tumour characteristics were similar in both groups. RESULTS: Right colonic obstruction or perforation was treated by primary resection and anastomosis in 11 of 12 patients before 1988 and in all 19 patients thereafter. Primary resection was also the treatment of choice for perforated cancer of the left colon and rectum before 18 of 20) and after (20 of 21) 1988. The rate of primary resection for obstructing cancer of the left colon and rectum increased from 17 of 45 to 30 of 35. One-stage resections for obstructing cancer were performed in ten of 45 and 22 of 35 patients before and after 1988 respectively. The overall mortality rate declined from 14 of 77 to three of 75 after 1988 (P< 0.01). The rate of radical lymphadenectomy rose from six of 46 patients to 42 of 69 after 1988. The 3-year survival rate increased from 50 to 74 per cent (P < 0.05). CONCLUSION: The data support further efforts towards improving the immediate and late outcome of emergency surgery in complicated colorectal cancer. PMID- 9752873 TI - Early experience with day-case transthoracic endoscopic sympathectomy. PMID- 9752874 TI - Malignant epithelial parotid gland tumours: analysis and results in 65 previously untreated patients. AB - BACKGROUND: Optimal management of malignant epithelial parotid tumours requires knowledge of the available therapeutic modalities and the different biological characteristics. The aim of the study was to review the characteristics of patients at presentation, histological classification, disease-free and overall survival rates, and the results of the applied treatment policy regarding the facial nerve and neck. METHODS: Between 1974 and 1995 a total of 65 patients was treated with curative intent for a previously untreated malignant epithelial parotid gland tumour. All patients underwent some type of parotidectomy, 20 of whom had an en bloc radical neck dissection. In selected cases the facial nerve or its branches were peeled off the tumour thus violating the objective of tumour free margins and relying heavily on the efficacy of postoperative radiotherapy. In total 51 patients received postoperative radiotherapy. None of the patients was lost to follow-up. RESULTS: There were 12 locoregional failures (18 per cent). In only one of these 12 patients was salvage therapy successful; the remaining 11 patients died from the tumour. All but one of the eight patients with distant metastasis only died from the tumour. The estimated 5- and 10-year disease-free rates were 68 and 59 per cent respectively. The corresponding survival rates were 75 per cent and 67 per cent. A significant relationship could be observed between tumour stage and survival. The presence of lymph node metastases proved to be the strongest single prognostic factor. CONCLUSION: In selected cases a conservative approach towards the facial nerve is justified. PMID- 9752875 TI - Detection of residual disease following breast-conserving surgery. AB - BACKGROUND: Assessment of completeness of tumour excision has become an integral part of breast-conserving surgery, but the accuracy of margin analysis has been questioned. This study compared the results of resection margin analysis with an examination of tumour bed biopsies and of the excised cavity wall. METHODS: One hundred and forty-four patients underwent breast-conserving surgery for T1-2 N0-1 breast cancer. Following wide local excision, four bed biopsies were taken from the cavity wall which was then completely excised. The presence of invasive and in situ disease at the inked resection margin (IRM) and in the adjacent bed biopsies and cavity wall was recorded. RESULTS: Positive margins and/or residual disease in either the bed biopsies or cavity wall was found in 62 (43 per cent) of 144 cases. Residual disease (invasive or in situ) was present at the IRM in 39 specimens (27 per cent) and was present in 25 bed biopsy (17 per cent) and 39 cavity wall (27 per cent) specimens. These comprised different but overlapping groups of patients. CONCLUSION: Margin analysis of wide local excision specimens is a poor predictor of completeness of excision. Routine resection and examination of the entire cavity wall increases the detection of residual disease compared with examination of bed biopsies alone and is a useful adjuvant to conventional margin evaluation. PMID- 9752876 TI - Efficacy of cetrimide-chlorhexidine combination in surgery for hydatid cyst. PMID- 9752877 TI - Informed consent in patients with acute abdominal pain. AB - BACKGROUND: This study aimed to establish perceptions of informed consent in patients undergoing surgery for acute abdominal conditions. METHODS: A prospective observational study was carried out using a structured questionnaire based interview technique in patients undergoing surgery for acute abdominal conditions. Main outcome measures were to establish the effects of pain and preoperative analgesia on informed consent, patient comprehension of planned surgery and the degree of discussion of potential side-effects and complications. RESULTS: Thirty-one of 49 patients perceived that pain did not interfere with their ability to give informed consent. Forty of 48 stated that preoperative analgesia did not impair their ability to give informed consent. Forty-two understood why an operation was being planned but 28 patients stated that there had been no preoperative discussion of any potential side-effects or complications of surgery. CONCLUSION: In this study the majority of patients perceived that they retained the ability to give informed consent despite the effects of pain and analgesia. Although the majority of patients understood why an operation was being planned there is a clear need for improved discussion of potential side-effects and complications. PMID- 9752878 TI - Perigastric lymph node status as a prognostic indicator in patients with gastric cancer. AB - BACKGROUND: The extent of lymph node dissection and histological examination of dissected lymph nodes varies among countries, which leads to the erroneous nodal stage and different surgical results in gastric cancer (stage migration, 'Will Rogers effect'). The aim of this study was to clarify the prognostic significance of the number of positive perigastric lymph nodes, which could be evaluated simply after D1 gastrectomy. METHODS: A consecutive series of 106 patients with histologically node-positive gastric cancer treated by radical gastrectomy and lymph node dissection (D2 or D3) was studied. The number of metastatic perigastric nodes (level I, nos 1-6) was examined, and its influence on the survival of patients was analysed. RESULTS: The overall 5-year survival rate was 50.9 per cent; the 5-year survival rate was significantly decreased when positive perigastric nodes exceeded six (62 per cent for one to six nodes versus 23 per cent for seven or more nodes, P< 0.001). Tumours having one to six positive perigastric nodes compared with those having seven or more positive perigastric nodes were more likely to have a size less than 4 cm (29 per cent versus one of 30, P< 0.001), grossly localized type (45 per cent versus seven of 30, P=0.042), absence of serosal invasion (32 per cent versus none of 30, P=0.002) and metastasis limited to the perigastric lymph nodes (70 per cent versus seven of 30, P < 0.001). CONCLUSION: The results indicate that the number of positive perigastric nodes correlates with tumour progression and patient survival. This parameter is a simple and useful prognostic indicator for node-positive gastric cancer, and is available not only for D2 and D3 gastrectomy but also for D1 gastrectomy. PMID- 9752879 TI - Gastric carcinoid expresses the gastrin autocrine pathway. AB - BACKGROUND: In gastric adenocarcinoma the gastrin autocrine-paracrine pathway is activated. As enterochromaffin-like (ECL) cells originate from the same stem as epithelial cells, the aim of this study was to determine if the gastrin autocrine pathway is present in gastric carcinoid. METHODS: Samples from ten patients with gastric carcinoid were assessed by immunocytochemistry using primary antibodies directed against gastrin precursors and the gastrin/cholecystokinin B receptor and detected using the avidin-biotin immunoperoxidase system. RESULTS: A high level of expression of precursor and mature gastrin peptides, together with the gastrin receptor, was seen in all carcinoids screened. CONCLUSION: In common with the glandular epithelium of the stomach the gastrin gene is activated during the neoplastic process in ECL cells. This finding may explain why some carcinoids do not regress after surgical procedures that lower serum gastrin. Antigastrin agents may be a useful treatment for carcinoid either in their own right or as an adjunct to surgery. PMID- 9752880 TI - Oesophageal dysmotility is not associated with poor outcome after laparoscopic Nissen fundoplication. AB - BACKGROUND: Nissen fundoplication has become the standard operation in the surgical management of gastro-oesophageal reflux disease. Postoperative dysphagia is thought to occur more commonly in patients with oesophageal dysmotility and it has been recommended that fundoplication be modified or avoided in these patients. The aim of this study was to determine the outcome of patients with normal motility and dysmotility undergoing laparoscopic Nissen fundoplication. METHODS: This was a single-centre prospective cohort study with 1-year follow-up, using dysphagia as the main outcome variable. Of 81 patients who underwent laparoscopic surgery, 48 had normal motility and 33 had oesophageal dysmotility (defined as percentage peristalsis, using ten wet swallows, of 50 per cent or less and/or a mean distal pressure of less than 40 mmHg). RESULTS: Dysphagia was present before operation in 14 of 48 patients with normal motility and 15 of 33 in the dysmotility group (P=0.2). At 3-month follow-up, new or worse dysphagia was present in 13 of 48 patients in the normal group and four of 33 in the dysmotility group (P=0.17). At 1 year the incidence of dysphagia was six of 48 in the normal group and five of 33 in the dysmotility group (P=0.9). CONCLUSION: Preoperative manometric assessment of oesophageal motility does not correlate with postoperative outcome, and oesophageal dysmotility should not be regarded as a contraindication to laparoscopic Nissen fundoplication. PMID- 9752881 TI - Natural history of adhesional small bowel obstruction: counting the cost. AB - BACKGROUND: The aim of this study was to identify patients admitted with adhesional obstruction to determine if there was an identifiable pattern to the type of initial operation, the type of treatment used for the obstructive episode and the subsequent need for further treatment. METHODS: Patients with adhesional obstruction were identified retrospectively in a cross-sectional study using ICD codes relating to admissions in the years 1990 to 1996. The case notes were used to assess their outcome. RESULTS: Fifty-nine case notes from a total of 175 identified initially satisfied the inclusion criteria. These patients had a mean age at presentation of 51 (range 16-88) years and had undergone a total of 122 operations. Thirty-one patients (53 per cent) had a single previous operation with a median time to presentation with obstruction of 5.5 years (range 11 days to 34.7 years); 33 patients (56 per cent) were treated conservatively on their first admission. There was no statistically significant difference in the outcome in patients who received either conservative or surgical treatment. The length of stay in patients treated surgically (median 11 (range 2-47) days) was significantly longer than that for those treated conservatively (median 6 (range 1-39) days) (P< 0.001). A flow chart was constructed demonstrating the eventual outcome of the patients in the study, enabling the cost of adhesional obstruction to be calculated. CONCLUSION: This type of approach could be used to assess the potential effect of different treatment strategies for adhesional obstruction. PMID- 9752882 TI - Laparoscopic drainage of liver abscesses. PMID- 9752883 TI - Effect of endoscopic sphincterotomy and interval cholecystectomy on late outcome after gallstone pancreatitis. PMID- 9752884 TI - Non-operative management of blunt hepatic trauma. PMID- 9752885 TI - Changing trends in the management of phaeochromocytoma. PMID- 9752886 TI - Randomized multicentre trial of the influence of recombinant human erythropoietin on intraoperative and postoperative transfusion need in anaemic patients undergoing right hemicolectomy for carcinoma. PMID- 9752887 TI - Outcome following laparoscopic resection for colorectal cancer. PMID- 9752888 TI - Paradoxical hypertension after reversal of heart failure in patients treated with intensive vasodilator therapy. AB - Hypertension is a major cause of heart failure, evolving from left ventricular hypertrophy to systolic and diastolic dysfunction. Although effective heart failure therapy has been associated with a lowering or no change in systemic arterial blood pressure in long-term follow-up, this study describes the symptomatic, clinical, and left ventricular functional response of a subgroup of heart failure patients with a prior history of hypertension who demonstrated a paradoxical hypertensive response despite high-dose vasodilator therapy. We prospectively identified 45 patients with a past history of hypertension who had become normotensive with symptomatic heart failure. Of these 45 heart failure patients, 12 became hypertensive while receiving therapy in follow-up, with systolic blood pressure > or = 140 mm Hg (Group A). The remaining 33 patients did not have a hypertensive response to therapy (Group B). In the 12 Group A patients, 60+/-10 years old, with symptomatic heart failure for 6.3+/-4.3 years, vasodilator therapy was intensified in the 2.0+/-0.5 years of follow-up, achieving final doses of enalapril 78+/-19 mg and isosorbide dinitrate 293 +/-106 mg per day. New York Heart Association classification improved from 2.9+/-0.8 to 1.3+/-0.5 (P < or = .0001), with a reduction in heart-failure-related hospitalizations. Left ventricular ejection fraction increased from 17+/-6% to 40+/-10% (P < .0001). Follow-up blood pressure at 1 to 3 months was unchanged. However, both systolic and diastolic blood pressure increased at final follow-up, rising from 116+/-14 to 154+/-13 mm Hg (P = .0001) and from 71+/-9 to 85+/-14 mm Hg (P = .004), respectively. Renal function remained unchanged. Although both groups had similar clinical responses, there were more blacks and women in the hypertensive Group A. Effectively, 12 of 45 (27%) heart failure patients with an antecedent history of hypertension demonstrated a paradoxical hypertensive response to vasodilator therapy. The recurrence of hypertension in a significant portion of patients successfully treated for heart failure has important clinical implications. PMID- 9752889 TI - Strict dietary sodium reduction worsens insulin sensitivity by increasing sympathetic nervous activity in patients with primary hypertension. AB - To assess the effects of sodium reduction on insulin sensitivity in hypertension, we examined the change of insulin sensitivity after two degrees of dietary sodium restriction by the euglycemic hyperinsulinemic glucose clamp method in 12 subjects with primary hypertension. A controlled period of 1 week, when the subjects were taking a normal sodium diet, was followed by a randomized crossover study in which the subjects were placed on either moderate or strict reduced sodium diets for 1 week. The result of the 1-week moderate dietary sodium reduction from 200 to 100 mmol/day showed significant decreases in systolic and diastolic blood pressure by 6.5 and 5.0 mm Hg, respectively. Strict dietary sodium reduction to 30 mmol/day for 1 week resulted in no further decrease in blood pressure, but it increased plasma insulin by 40.6% without changing plasma glucose. There were no changes in glucose infusion rate (GIR) or insulin sensitivity index (ISI), which is a measure of GIR divided by plasma insulin, after moderate dietary sodium reduction. However, strict dietary sodium reduction induced decreases in GIR by 19.8% (from 1318+/-189 to 1057+/-173 micromol/m2/ min; P < .01), and ISI by 20.5% (from 16.6+/-2.1 to 13.2+/-1.9 micromol/m2/min/microU/mL; P < .01) with a paralleled increase of plasma norepinephrine by 90.0% (from 150.5+/-61.6 to 287.3+/-114.9 pg/mL; P < .01). These results indicate that dietary sodium restriction leads to a deterioration of insulin sensitivity when plasma norepinephrine levels increase, and suggest that moderate dietary sodium reduction may lower blood pressure without a distinct adverse effect on glucose metabolism in subjects with primary hypertension. PMID- 9752890 TI - Effects of aging on the insulin actions for the glucose metabolism and renal function in normotensives and essential hypertensives. AB - It has been suggested that hyperinsulinemia compensating insulin resistance in glucose metabolism may be a pathogenic factor in essential hypertension. On the other hand, age-associated increases in the prevalence of glucose intolerance and hypertension are also well established. The aim of this study is to clarify the influence of aging on insulin sensitivity in glucose metabolism and on renal sodium handling under hyperinsulinemia, which may relate to high blood pressure in insulin-resistant subjects. Fifty-two normotensive subjects and 61 patients with essential hypertension were evaluated in this study. The subjects of these groups were divided into young (<40 years old) and middle-elderly (> or = 40 years old): young normotensives (Y-NT, n = 22); middle-elderly normotensives (ME- NT, n = 30); young hypertensives (Y-HT, n = 9); and middle-elderly hypertensives (ME-HT, n = 52). Using the euglycemic hyperinsulinemic glucose clamp, insulin sensitivity was assessed as M value. Just before the start and the termination of the glucose clamp, creatinine clearance (Ccr) and urinary excretion of sodium (UNaV) were measured. In addition, renal plasma flow assessed as para aminohippuric acid clearance was also measured at the same time in several subjects; 8 Y-NT, 8 ME-NT, 3 Y-HT, and 10 ME-HT. The M value was significantly lower in ME-NT, Y-HT, and ME-HT, compared to Y-NT, although blood sugar and immunoreactive insulin levels were similar in all four groups. In normotensive subjects, there was a significant, negative correlation between age and M value. However, this correlation was not observed in hypertensive patients. UNaV decreased in ME-NT, Y-HT, and ME-HT, but not in Y-NT under hyperinsulinemia by the glucose clamp, whereas Ccr showed no significant change in any group. In all subjects, the change of UNaV (deltaUNaV) correlated significantly and positively with the M value. Renal plasma flow significantly increased under hyperinsulinemia by the glucose clamp in only Y-HT, but not in the other groups. There was a significant, positive correlation between deltaUNaV and the change of renal plasma flow under hyperinsulinemia by the glucose clamp. These results suggested that both the impairments of the insulin sensitivity and insulin induced vasodilation at the renal artery with aging may partially contribute to age-related elevation of blood pressure through renal sodium retention by compensating hyperinsulinemia. On the other hand, it seems reasonable to assume that these abnormalities, which can contribute to high blood pressure in essential hypertension, already may exist at lower ages in essential hypertensive patients. PMID- 9752891 TI - Ambulatory blood pressure and urinary albumin excretion in diabetic (non-insulin dependent and insulin-dependent) hypertensive patients: relationships at baseline and after treatment by the angiotensin converting enzyme inhibitor trandolapril. AB - The aim of the present study was to examine the relationships between ambulatory blood pressure (ABPM) and urinary albumin excretion (UAE) in diabetic (non insulin dependent [NIDDM] and insulin-dependent [IDDM]) hypertensives at baseline and after treatment by an angiotensin converting enzyme (ACE) inhibitor. After a 3-week placebo period, patients were treated for 16 weeks with trandolapril, 2 to 4 mg/day. The UAE and blood pressure (mercury sphygmomanometer and 24-h ABPM) were measured at baseline and repeated on trandolapril. Predictive factors of abnormal UAE (24-h UAE > or = 30 mg) were determined using univariate and multivariate analysis (logistic regression). Predictors of UAE decrease were also searched. One hundred seventy-one patients entered the analysis. Baseline office BP was 164+/-14 / 97+/-6 mm Hg and 24-h BP was 142+/-17 / 83+/-10 mm Hg. Seventy four patients (43%) had UAE > or = 30 mg. Independent risk factors for abnormal UAE were nighttime diastolic BP (odds ratio [OR] = 4.1, confidence interval [CI] = 2.0 to 8.6, P = .0001), diabetes duration (OR = 2.4, CI = 1.1 to 5.0, P = .025), and presence of retinopathy (OR = 3.2, CI = 1.0 to 10.0, P = .047). Conversely, office BP level was not significantly related to UAE. On treatment, office BP levels decreased to 143+/-13 / 82+/-8 mm Hg (P < .0001) and 24-h BP levels to 134+/-17 / 78+/-9 mm Hg (P < .0001). In the abnormal UAE group, UAE significantly decreased from 76 to 50 mg/day (P = .006). After treatment, independent predictive factors of abnormal UAE were: on-drug fasting plasma glucose (OR = 3.5, CI = 1.7 to 7.4, P = .0009) and on-drug nighttime diastolic BP (OR = 3.5, CI = 1.7 to 7.4, P = .001). The only predictor of UAE decrease was a 24-h systolic BP decrease (OR = 2.3, CI = 1.3 to 4.3, P = .007). We conclude that in diabetic hypertensives with abnormal UAE, trandolapril exhibited a sustained 24-h antihypertensive effect and provided a consistent reduction of microalbuminuria. This study confirmed the superiority of ABPM over clinical BP to predict target organ damage. PMID- 9752893 TI - Long-term follow-up of acute renal failure caused by angiotensin converting enzyme inhibitors. AB - Angiotensin converting enzyme (ACE) inhibitors are useful in the treatment of hypertension and heart failure. However, acute renal failure (ARF) may occur in patients who are taking these drugs in situations associated with decreased glomerular filtration pressure, such as dehydration caused by acute diarrhea or diuretic therapy. Sixty-four patients who were admitted to the intensive care unit for ARF associated with ACE inhibitor therapy were followed for more than 5 years. In this historical retrospective study, we documented that 45 patients were treated for hypertension (group I) and 19 were treated for heart failure (group II). Their mean age was 71.2+/-11.6 years. Patients with ARF presented with overt dehydration in 91% and 84% of the cases in groups I and II, respectively. Hypovolemia was caused by diuretics or gastrointestinal fluid loss. Bilateral artery-renal stenosis or stenosis in a solitary kidney was documented in 22% and 10% of patients in groups I and II, respectively. The probability of survival was 91% and 49% at 1 year and 64% and 18% at 5 years, for groups I and II, respectively. Acute renal failure required hemodialysis in seven patients, but none of them became dialysis dependent. In the subgroup of patients with preexisting chronic renal failure, all the patients except for one who belonged to group II died within 2 years. In both groups, after resolution of ARF, plasma creatinine concentration returned to baseline level and the course of renal function was not significantly worsened. In conclusion, ARF associated with ACE inhibitors is likely to occur in many patients without renal artery stenosis after unexpected dehydration, especially in older patients with congestive heart failure. In both groups of patients, in the absence of preexisting chronic uremia, recovery of renal function occurred without sequelae, even after an episode of acute tubular necrosis requiring dialysis. PMID- 9752892 TI - Comparison between the effects of amlodipine and lisinopril on proteinuria in nondiabetic renal failure: a double-blind, randomized prospective study. AB - Double-blind, randomized controlled studies of longer than 1 week in duration comparing the antiproteinuric potential of long-acting dihydropyridine calcium channel blockers with that of angiotensin converting enzyme (ACE) inhibitors are lacking. Therefore, we performed such a study in patients with nondiabetic renal disease and proteinuria. After a 4-week wash-out period in which patients did not use any medication known to affect proteinuria, 21 patients were randomized in a double-blind fashion to receive either the calcium channel blocker amlodipine (Amlo, 5 to 10 mg) or the ACE-inhibitor lisinopril (Lis, 5 to 10 mg). Throughout the 16-week study period, blood pressure, creatinine clearances, and proteinuria were measured every 2 weeks. In addition, device-measured blood pressure and renal hemodynamic studies were performed at the start and end of the study. Systolic blood pressure fell in the Lis group from 163+/-7 (SEM) to 140+/-8 mm Hg (P < .01) and from 157+/-10 to 147+/-6 mm Hg in the Amlo group; diastolic blood pressure fell from 101+/-3 to 86+/-7 mm Hg in the Lis group and from 98+/-3 to 91+/-2 mm Hg in the Amlo group. Renal hemodynamics were not affected by amlodipine treatment, whereas a fall in glomerular filtration rate (GFR) was seen in lisinopril-treated patients (from 55+/-11 to 50+/-10 mL/min; P < .01). Amlodipine did not significantly affect proteinuria. Lisinopril induced a decline in the protein-creatinine ratio with a maximal effect reached after 12 to 16 weeks of therapy (from 0.39+/-0.17 to 0.26 +/-0.11 g/mmol; P < .009). In conclusion, we could not demonstrate an antiproteinuric effect of the long-acting dihydropyridine calcium channel blocker amlodipine, whereas therapy with the ACE inhibitor lisinopril resulted in a decrease in proteinuria. Amlodipine did not affect renal hemodynamics, whereas lisinopril induced a fall in GFR. PMID- 9752894 TI - Fixed low-dose perindopril-indapamide combination in hypertensive patients with chronic renal failure. AB - The angiotensin converting enzyme inhibitor perindopril and the diuretic indapamide have been shown to be effective antihypertensive agents in patients with chronic renal failure. A fixed low-dose combination of these two agents has been proposed in the treatment of hypertension. We evaluated this combination in 26 patients with mild to moderate essential hypertension and mild to severe chronic renal failure that did not require dialysis. This was a multicenter, open trial consisting of a 2-week single-blind placebo washout period followed by 12 weeks of active treatment. At week 0, the patients received 2 mg perindopril/0.625 mg indapamide once a day or every other day, with the possibility of dosage adjustment to perindopril 4 mg/indapamide 1.25 mg at week 2, week 4, or week 8. A pharmacokinetic analysis using a population pharmacokinetic approach was performed at week 8. Twenty-three patients completed the 12-week study, at which time 14 patients were receiving 2 mg perindopril/0.625 mg indapamide daily, three were receiving 2 mg perindopril/0.625 mg indapamide every other day, and six perindopril 4 mg/indapamide 1.25 mg. Blood pressure readings (supine) decreased from 170.4+/ 19.2 / 101.5+/-6.7 mm Hg before active treatment to 146.5+/-19.7 / 86.5+/-10.6 mm Hg at the end of treatment (P < .0001). Pharmacokinetic analysis showed that for indapamide and perindoprilat (the active metabolite of perindopril) the area under the curve (AUC24) increased with the severity of renal failure. No interaction was noted between the two drugs. Mean serum creatinine and sodium and serum potassium levels remained stable during the study. Impairment of renal function occurred in one patient and was considered unrelated to treatment. We conclude that a fixed low-dose perindopril-indapamide combination as first-line treatment has a good safety/efficacy ratio in hypertensive patients with chronic renal failure. PMID- 9752895 TI - Effects of smoking on 24-h ambulatory blood pressure and autonomic function in normoalbuminuric insulin-dependent diabetes mellitus patients. AB - Smoking is an important risk factor for the development and progression of diabetic nephropathy. The mechanisms by which smoking increases albuminuria and promotes nephropathy are unknown. Considering the acute pressor effect of smoking and the close association between blood pressure elevation and development of diabetic nephropathy, blood pressure increase might be implicated in the association between smoking and diabetic nephropathy. However, among nondiabetics, smokers have repeatedly been found to have lower blood pressure than nonsmokers. This is possibly mediated by an autonomic adjustment to sustained sympathetic stimulation by nicotine. Impaired modulation of the sympathovagal activity has been described in diabetes. In diabetic patients, the effect of smoking on blood pressure and autonomic function remains unclarified. We examined 24-h ambulatory blood pressure (oscillometric technique) and autonomic function (short-term power spectral analysis as well as conventional tests) in 24 smokers and 24 nonsmokers matched for sex, age, and diabetes duration. All patients were normoalbuminuric insulin-dependent diabetes mellitus patients. Smoking status was assessed by questionnaire with confirmatory determinations of urinary cotinine. Diabetic smokers had significantly higher 24 h mean arterial blood pressure (94+/-6.7 mm Hg compared to diabetic nonsmokers 90+/-5.8 mm Hg, P = .04) including higher diastolic nighttime blood pressure (68+/-7.3 mm Hg v 64+/-5.2 mm Hg, P = .03). Smokers also had significantly higher 24-h heart rate (80+/-7.2 compared to 72+/-9.2 beats/min, P < .001). In addition, smoking was associated with significantly reduced short-term RR interval variability (supine low frequency component) (5.45+/-1.29 ln ms2 in smokers compared to 6.31+/-1.11 ln ms2 in nonsmokers, P < .02), as well as reduced brake index (33.5+/-14.5 in smokers v 42.1+/-16.0 in nonsmokers, P < .05). Diabetic smokers have significantly higher 24-h blood pressure compared to diabetic nonsmokers. This finding, contrasting the effect of smoking among nondiabetics, is possibly mediated by coexisting abnormal postural responses in autonomic cardiac regulation in diabetic smokers. Blood pressure elevation, persisting throughout 24 h, might be operative in the association between smoking and development of diabetic nephropathy. PMID- 9752896 TI - Plasma homocysteine and folate are related to arterial blood pressure in type 2 diabetes mellitus. AB - The aim of this study was to assess the relationship between homocysteine (tHcy), folate and vitamin B12 levels, urinary albumin excretion, and arterial blood pressure in patients with non-insulin-dependent diabetes mellitus (NIDDM). Our study was carried out in 33 NIDDM patients (16 men, 17 women) and 16 healthy volunteers as controls (seven men, nine women). Fasting and postmethionine load plasma tHcy levels were assessed, together with folate, vitamin B12, and urinary albumin excretion levels. In NIDDM patients, there were correlations between folate and mean arterial pressure (r = -0.352, P = .046), folate and systolic blood pressure (r = -0.437, P = .013), folate and vitamin B12 (r = 0.499, P = .004), tHcy and vitamin B12 (r = -0.348, P = .04), ln tHcy and ln folate (r = 0.404, P = .01), and, lastly, between tHcy, either fasting or postload, and urinary albumin excretion. Patients with elevated tHcy levels had significantly higher diastolic blood pressure (P = .04) and mean arterial pressure (P = .03). Otherwise, higher folate values were associated with lower systolic blood pressure (P = .004) and mean arterial pressure (P = .02). In addition, NIDDM patients with complications presented higher tHcy basal values than the group without complications (P = .003). A particular propensity of such patients towards endothelial dysfunction could explain the presence of correlations between these metabolic parameters and arterial blood pressure. PMID- 9752897 TI - Apoptosis in vasculature of spontaneously hypertensive rats: effect of an angiotensin converting enzyme inhibitor and a calcium channel antagonist. AB - Increased apoptosis has been demonstrated in various forms of human and experimental cardiovascular disease. The role of this phenomenon in the vasculature in different models of hypertension is unclear. In hypertension, regression of vessel wall hypertrophy/hyperplasia or remodeling in response to various antihypertensive drugs may be mediated in part by apoptosis. This study examined vascular smooth muscle apoptosis in spontaneously hypertensive rats (SHR), in which it may presumably counterbalance vascular wall growth. Angiotensin converting enzyme (ACE) inhibitors and calcium channel blockers induce regression of the vascular wall in hypertension. Therefore, we investigated the effect of the ACE inhibitor enalapril and the dihydropyridine calcium channel blocker amlodipine on apoptosis in blood vessels of SHR to determine whether part of the growth inhibitory effect of these drugs is mediated by apoptosis. This was performed by detection and measurement of DNA fragmentation using DNA laddering and examining aortic histologic sections with in situ end-labeling (terminal deoxynucleotide transferase-mediated dUTP-nick labeling [TUNEL]). Ten-week-old SHR were treated for 12 weeks with 10 mg/kg per day of enalapril and 20 mg/kg per day of amlodipine. Blood pressure was significantly reduced by enalapril and amlodipine (P < .01). Cross-sectional area of aorta was significantly increased (3.34+/-0.15 mm2) in SHR compared to that of Wistar-Kyoto (WKY) control rats (1.17+/-0.07 mm2, P < .01). The cross-sectional area of the aorta was significantly smaller in enalapril-treated SHR (2.42+/-0.12 mm2, P < .05) compared to untreated SHR, and almost normalized by amlodipine (1.65+/-0.31 mm2, P < .01). Apoptosis characterized using terminal deoxynucleotidyl transferase to radiolabel 3'-OH ends of fragmented DNA extracted from aorta, showed presence of fragmented labeled DNA as "DNA laddering," a hallmark of apoptosis. SHR had increased apoptosis (341+/-25 pixels/microg DNA) compared to WKY controls (206+/-13 pixels/microg DNA, P < .01). Apoptosis was six to eightfold greater in aorta of enalapril and amlodipine-treated SHR (P < .01). These results were confirmed by in situ end-labeling of fragmented DNA in aortic histologic sections. Western blot quantification of Bax and Bcl-2 (pro- and antiapoptotic gene products, respectively) showed higher Bax and lower Bcl-2 expression, and accordingly increased the Bax-to-Bcl-2 ratio in aorta from SHR treated with enalapril or amlodipine in comparison to untreated SHR. In conclusion, enhanced apoptosis is present in aorta of SHR, possibly as a homeostatic mechanism counterbalancing growth. Antihypertensive agents such as the ACE inhibitor enalapril and the calcium antagonist amlodipine may cause regression or inhibition of vascular wall growth in SHR partly through enhanced apoptosis, which may contribute to the antihypertensive effects of these drugs. PMID- 9752898 TI - The perivascular sensory nerve Ca2+ receptor and blood pressure regulation: a hypothesis. PMID- 9752899 TI - Renal artery clipping attenuates the progression of adriamycin nephropathy. AB - This study was designed to analyze the impact of diminished renal perfusion pressure due to renal clipping on the rat model of adriamycin nephropathy. Male Wistar rats, divided into four groups (n = 9 per group) were injected with saline as control (C), adriamycin 3 ml/kg (Ad), saline with the left renal artery clipped (Rv), and adriamycin plus clip (AdRv). After 24 weeks mean arterial pressure (MAP), inulin, and p-aminohippurate (PAH) clearances were performed to evaluate renal function. Morphologic analysis included histologic criteria of percentage of glomerulosclerosis and tubulointerstitial lesion index (TILI). The MAP (mm Hg) was similar between Rv (143+/-13) and AdRv (154+/-20), but higher (P < .05) than C (120 +/-8) and Ad (124+/-11). Inulin clearance (mL/min/ 100 g) in Ad (0.2+/-0.05) was smaller than in C (0.53+/-0.17) and Rv (0.4+/-0.01) (P < .05), and was at an intermediate level in AdRv (0.33+/-0.2). The level of PAH (mL/min/100 g) was normal at 1.76 in C, and diminished more in Ad (0.58) than in Rv (1.06) and AdRv (1.18) (P < .05). Both Ad and the AdRv nonclipped kidneys had the highest degree of glomerulosclerosis (33% and 25%) and TILI (7% and 8%), respectively, compared with C and Rv (both 0%), whereas the clipped kidneys displayed intermediate degrees (9% and 5%) (P < .05 v nonclipped). The data suggest that diminished perfusion pressure of the clipped kidney, by decreasing the intraglomerular pressure, protects the glomerulus from damage and attenuates the evolution of adriamycin nephropathy. PMID- 9752900 TI - Chronic hypertension leads to hyperinsulinemia in Sprague-Dawley rats treated with nitric oxide synthase inhibitor. AB - Insulin resistance and hypertension, as well as dyslipidemia, frequently cooccur. Evidence that nitric oxide (NO) plays a crucial role in the long-term regulation of systolic blood pressure led us to examine whether enhanced vasoconstriction and hypertension induced by NO synthase inhibitor could lead to insulin and lipid disorders. NG-Nitro-L-arginine methyl-ester (L-NAME), an inhibitor of NO synthase, was given for 4 weeks in drinking water (100 mg/kg/day) to 12 Sprague Dawley rats. Another nine rats received both L-NAME and verapamil (100 mg/kg/day), whereas 12 animals fed rat chow only served as controls. Systolic blood pressure was measured weekly by the indirect tail cuff method. Blood samples were taken at the beginning of the experiment, and after 2 and 4 weeks from all rats. The samples were assayed for insulin, glucose, and triglyceride concentrations. L-NAME treatment resulted in a marked and sustained increase in systolic blood pressure from 130+/-7 to 171+/-3 mm Hg by the second week, which was succeeded by a significant elevation in insulin level at the end of 4 weeks, from 2.3+/-1.8 to 5.4+/-2.0 ng/mL. Triglycerides and glucose were unaffected throughout the experiment. The combination of L-NAME and the NO-independent vasodilator, verapamil, attenuated the hypertension induced by L-NAME and prevented the following rise in insulin level. Data suggest that chronic elimination of NO after chronic inhibition of NO synthase may lead to a state of hyperinsulinemia, possibly as an outcome of insulin resistance. PMID- 9752901 TI - Troglitazone, an insulin sensitizer, increases forearm blood flow in humans. AB - To test whether troglitazone, a thiazolidinedione insulin sensitizer, increases the peripheral blood flow, the changes in forearm blood flow (FBF) were evaluated by venous occlusion plethysmography in 11 lean healthy male volunteers (age range, 24 to 39 years) after a single oral dose of 200 mg of troglitazone. Forearm vascular resistance (FVR) was calculated from FBF and blood pressure. Two hours after the dose, FBF increased from 3.66+/-0.31 to 4.81+/-0.57 mL/100 mL/min (P < .01), and FVR decreased from 24.7+/-2.2 to 20.2+/-2.2 units (P < .01), whereas both these values did not change during the control recordings obtained without troglitazone. Blood pressure, blood glucose levels, and serum immunoreactive insulin levels did not change significantly during the observation period. Serum concentrations of nitrate ions decreased from 27.0+/-3.5 mmol/L to 23.1+/-2.7 mmol/L (P < .01) after the administration. These results suggest that troglitazone increases muscular blood flow through vasodilation induced by a mechanism other than the correction of hyperinsulinemia or the increase in nitric oxide. The present study provides the first evidence that troglitazone dilates the vasculature in humans. PMID- 9752902 TI - Management of high casual blood pressure in a disaster situation: the 1995 Hanshin-Awaji earthquake. PMID- 9752903 TI - The virtual hand. The Pulvertaft Prize Essay for 1996. AB - Virtual reality technologies are now at a stage in which the various disciplines can be brought together to construct a virtual human hand. Devices can be constructed to record multiple joint positions accurately in clinical environments. Joint prostheses may be tested virtually before undergoing clinical trials, albeit in a simple way at present, but may eventually be incorporated into a virtual model of the hand and driven by goniometric gloves. This will allow more detailed analyses of implant in situ behaviour. These exciting developments will provide a huge advance in our understanding of the functions of the real hand and also a potential way of assessing outcomes in a simple and repeatable fashion. We are on the edge of a new era in hand surgery when the computer scientist, biomechanic, control engineer, hand therapist and surgeon will be able to alternate between the virtual and the real world in producing better outcomes for patients. PMID- 9752904 TI - Side-effects of rinsing fluids on the rat femoral artery. A histological and electron microscopic study. AB - Rinsing rat femoral arteries with various fluids in experimental conditions similar to those in clinical practice was found to have deleterious effects on the intimal and medial layers of the vessels. No statistically significant difference was found between the effects of Ringer's lactate and normal saline. Heparinized saline produced significantly less damage to the medial layer and less platelet cell deposition. Lignocaine 2% was found to be extremely damaging to the whole vessel wall, and highly thrombogenic. Nevertheless, all the arteries in each group remained patent 4 days after rinsing. PMID- 9752905 TI - Portals for arthroscopy of the trapeziometacarpal joint. PMID- 9752906 TI - Trapeziometacarpal joint arthrodesis for osteoarthritis. Results of power staple fixation. AB - Fifty-two cases of trapeziometacarpal joint osteoarthritis were treated with arthrodesis using small staple fixation. Eighty-five per cent of the patients were free of pain, with normal thumb. Opposition to the fourth finger tip was always possible and opposition to the little finger tip was observed in 94% of cases. Four patients (7.6%) developed a non-union but only two complained of pain. Arthrodesis with power staple fixation minimizes bone resection and postoperative immobilization (mean 28 days) because of the good stability following this procedure. PMID- 9752907 TI - Expandable rings. A new solution to an old problem. AB - A number of solutions are presented for the problem of over-tight rings particularly in patients with deforming arthropathies of the hand. Rings can be adapted by various methods to allow them to be worn on deformed fingers. An illustrative case is reported. PMID- 9752908 TI - Comparison between single injection transthecal and subcutaneous digital blocks. AB - A randomized double blinded study was performed on 142 patients to evaluate two different techniques of single injection digital anaesthesia. In group A, 86 digits in 71 patients were anaesthetized by a single injection transthecal technique using 3 cc of lignocaine and bupivacaine mixture. Anaesthesia of the whole digit was achieved in 83 (97%) digits. In group B, 80 digits in 71 patients were anaesthetized with a single injection subcutaneous technique using the same amount of anaesthetic mixture. Total anaesthesia of the digit was achieved in 75 (94%) digits. These two techniques were found to have no differences in effectiveness, distribution, onset and duration of anaesthesia. PMID- 9752909 TI - The extensor retinacular system at the metacarpophalangeal joint. Anatomical and histological study. AB - The anatomy of the sagittal bands was studied in 56 cadaver digits. The sagittal band is part of an extensor retinacular system which is integrated with the extrinsic and intrinsic musculotendinous structures. The extensor retinacular system is a single unit with radial and ulnar components and has transverse, sagittal and oblique fibres. The transverse-sagittal fibres, along with the palmar plate, form a closed cylindrical tube which surrounds the metacarpal head. The oblique fibres form the triangular lamina distal to the sagittal band. The radial component of the sagittal band is often thinner and longer than the ulnar component. The sagittal band envelops the extensor digitorum tendon and the superficial fibres are thinner than deep fibres, especially in the central digits. The central digits have palmar soft tissue confluence on each side consisting of the sagittal band, palmar plate, annular pulley and deep transverse metacarpal ligament. The sagittal band also appears to envelop the superficial interosseous tendons on both sides. Our findings explain the propensity for radial sagittal band injuries and suggest that the sagittal band is the primary stabilizer of the extensor digitorum at the metacarpophalangeal joint. PMID- 9752910 TI - Combined Sever's release of the shoulder and osteotomy of the humerus for Erb's palsy. AB - In children with Erb's palsy who do not recover fully, the most common residual deformity is internal rotation-adduction contracture of the shoulder. Surgical correction of this contracture is described in this paper in 12 consecutive patients by combined Sever's release of the shoulder and osteotomy of the humerus. After an average follow-up of 3 years, the average gain of active external rotation of the shoulder was 32 degrees and average gain of active abduction was 61 degrees. PMID- 9752911 TI - Early active mobilization for extensor tendon injuries. The Norwich regime. AB - Dynamic splinting following extensor tendon repair gives better results than static splinting, but involves cumbersome splints and recommended protocols are often complicated. We prefer controlled active mobilization of extensor tendon repairs without dynamic splinting. Six weeks after repair, excellent or good function was obtained in 22 out of 24 simple extensor tendon injuries and in 11 out of 13 complex injuries. The results of this prospective study are comparable with those reported after dynamic splinting; this regime does not require outrigger splintage and is simple to follow. PMID- 9752912 TI - Extensor tendon rupture due to Kienbock's disease. AB - Extensor tendon rupture as a complication of Kienbock's disease is rare. We report a case of attritional rupture of the extensor tendon to the index finger by a comminuted dorsal fragment of the lunate after Kienbock's disease in an elderly woman. Excision of the fragment and extensor reconstruction with tendon graft led to a favourable result. PMID- 9752913 TI - Carpal tunnel syndrome: neurophysiological results of surgery based on preoperative electrodiagnostic testing. AB - Fifty-three hands with carpal tunnel syndrome had pre- and postoperative evaluations of median nerve distal motor latency (from wrist to thenar muscles) and orthodromic sensory nerve conduction velocity (from thumb and middle finger to wrist). At 6 months we observed a neurophysiological return to normal in all cases with normal preoperative distal motor latency and in about 50% of the hands with preoperative distal motor latency between 4 and 6 ms. Prolongation of the distal motor latency over 6 ms was not followed by return to neurophysiological normality, although some degree of sensory function was restored in the majority of cases. PMID- 9752915 TI - Intrauterine vascular deficiency of the upper limb. AB - We report six cases of intrauterine vascular deficiency of the upper limb, presenting over a 12-year period, with established areas of necrosis at the time of birth. Three of the six mothers were diabetic. An urgent examination of the neonate should be done to exclude occult systemic thomboses, as early anticoagulation or thrombolytic therapy may be indicated. The majority of cases were managed conservatively with dressings and splintage of the limb; exceptionally surgical intervention was required. Long-term sequelae included flexion contractures and shortening of the forearm bones. PMID- 9752914 TI - Sensory function after median nerve decompression in carpal tunnel syndrome. Preoperative vs postoperative findings. AB - The sensory recovery was monitored for up to 1 year after decompression of the median nerve in 69 patients with carpal tunnel syndrome. Special attention was paid to the rate of recovery, the importance of constant or intermittent numbness or paraesthesiae preoperatively and the influence of gender. Most patients with numbness/paraesthesiae and those with abnormal two-point discrimination recovered within 10 days. Perception of touch and vibration recovered within 3 weeks in most patients but those with abnormal nerve conduction/sensory amplitude recovered slowly during follow-up. After 1 year patients with intermittent preoperative symptoms were significantly more likely to achieve normal nerve conduction and perception of touch. Women were more likely to achieve normal nerve conduction and perception of touch. A comparison of recovery between matched men and women with identical preoperative status showed no significant difference. The results indicate the importance of early treatment of carpal tunnel syndrome. PMID- 9752916 TI - Neonatal compression ischaemia of the forearm. AB - A case of neonatal compartment syndrome of the forearm is reported. The cause was thought to be compression during delivery. It not only caused muscle contractures but also affected bone growth. Conservative treatment was given. At the age of 6 years, the muscle contracture had recovered and there was full hand function, but there was growth arrest of the distal radial epiphysis and the affected forearm was shorter than the other. PMID- 9752917 TI - "Spare part" forearm free flaps harvested from the amputated limb for coverage of amputation stumps. AB - The use of "spare part" flaps from a non-replantable limb to cover amputation stumps in the upper extremity preserves limb length, provides durable coverage and sensation and will avoid additional donor site morbidity. We have studied the blood supply of the forearm based on the radial artery. The potential for harvesting different tissues is confirmed. In five clinical cases reliable primary soft tissue reconstruction was achieved, even in the presence of trauma. PMID- 9752918 TI - The combined use of a pedicled Scarpa's fascia flap and a groin flap for simultaneous coverage of dorsal and palmar finger defects. AB - A case is reported in which the dorsal and palmar aspects of the fingers in a severely crushed hand were covered by combining a pedicled Scarpa's fascia flap and a groin flap. Secondary heterotopic finger transposition was additionally performed to restore satisfactory hand function. An acceptable result was obtained. PMID- 9752919 TI - Reconstruction of large palmar defects of the hand using free flaps. AB - The reconstruction of large palmar defects of the hand remains a difficult problem due to the specific anatomical structures and highly sophisticated function of the palm. The glabrous skin and subcutaneous tissue in the palm are perfectly adapted to serve the prehensile function. The particular aim must be that repairs to this functional structure are similar in texture and colour and are aesthetically acceptable. Restoration of sensibility is desirable. For smaller defects a great variety of local pedicled or island flaps can be applied. However, for larger defects with exposed tendons, nerves or other essential structures, free flaps remain as a reliable alternative. This paper reviews our approach of soft tissue reconstruction in 16 patients with large palmar defects using various kinds of free flaps. The advantages, disadvantages and current indications for free flap resurfacing of the palm are discussed. PMID- 9752920 TI - The role of metacarpophalangeal pattern (MCPP) profile analysis in the treatment of triphalangeal thumbs. Description of a method and a case report. AB - The metacarpophalangeal pattern (MCPP) profile analysis is a method of comparing the length of each of the 19 tubular bones of the hand on the X-ray with the standard length in the normal population according to age and sex. An MCPP plot is a graphic illustration of the MCPP analysis. It is not the exact height of the curve on the MCPP plot which is most important, but the profile which occurs because of the individual lengthening or shortening of the bones. This pattern profile appears to be specific for several congenital malformation syndromes. We have recently used MCPP analysis in planning surgery for triphalangeal thumbs. The percentage of excessive or reduced length of each individual bone of the hand can be read from the MCPP plot and is helpful in calculating a more accurate length for the newly created thumb. MCPP analysis can be used as a diagnostic tool in a number of congenital hand malformations, but may also be helpful in planning surgical treatment of congenital hand malformations when abnormal bone length is involved. PMID- 9752921 TI - Conservative management of wrist ganglia. Aspiration versus steroid infiltration. AB - Wrist ganglia were randomly allocated for conservative treatment by either aspiration or aspiration and injection of steroid. Both treatment methods had 33% success rates. Almost all ganglia which recurred after one aspiration did not resolve with further aspirations. After aspiration and explanation of the benign nature of ganglia, only a quarter of patients requested surgical treatment. PMID- 9752922 TI - Intraarticular lesions in distal fractures of the radius in young adults. A descriptive arthroscopic study in 50 patients. AB - We examined the frequency of associated chondral and ligament lesions in distal fractures of the radius in young adults (men 20-60 years, women 20-50 years). Fifty initially displaced fractures were examined arthroscopically. Chondral lesions were found in 16 patients (32%). All patients but one were found to have a ligamentous injury in the wrist. No major instability was found. The most frequent ligament tear was the triangular fibrocartilage complex in 39 cases (78%), with a statistical correlation to ulnar styloid fractures. The scapholunate ligament was partially or totally torn in 27 cases (54%). No correlation was found between specific fracture type and pattern of ligament injury. Chondral and ligamentous lesions were frequent and may explain poor outcomes after seemingly well-healed distal fractures of the radius. The ligament lesions should also be kept in mind when early mobilization of the distal fracture of the radius is considered. PMID- 9752923 TI - The anatomical basis of the vascularized pronator quadratus pedicled bone graft. AB - The vascular supply of the pronator quadratus was studied in 25 cadaveric dissections following coloured latex injections. This showed that the main blood supply of the pronator quadratus came from the anterior interosseous artery. There was, however, a rich anastomosis between the branches of the anterior interosseous artery and those of the radial and ulnar arteries. It was possible to raise a corticocancellous bone graft from the anterior surface of the radial styloid on a pedicle of the lower fibres of the pronator quadratus muscle. This muscle pedicle had a constant branch of the anterior interosseous artery which vascularized the bone graft. Such a vascularized pedicled bone graft may be useful in the treatment of non-union of the scaphoid and Kienbock's disease. PMID- 9752924 TI - Management of acute perilunate dislocations without fracture of the scaphoid. AB - A retrospective review of 14 cases of acute perilunate dislocations without fracture of the scaphoid managed by three different forms of treatment was conducted at an average follow-up of 29 months. Treatment included closed or open reduction with cast immobilization only (n=2), closed reduction followed by percutaneous K-wire fixation of the carpus (n=4), and open reduction with repair of the torn scapholunate ligaments and K-wire fixation of the carpus (n=8). Based on Cooney's clinical scoring system, there were five excellent, five good, two fair and two poor results. The patients without ligamentous repair did as well as those with ligamentous repair when the scaphoid was reduced anatomically and stabilized with K-wires. In the latter, however, the scapholounate relationship was maintained more consistently. We believe that open reduction through a dorsal approach, direct repair of the scapholunate ligaments, and K-wire fixation of the carpus is a reliable method for obtaining satisfactory clinical and radiographic results in the management of acute perilunate dislocations without fracture of the scaphoid. PMID- 9752925 TI - Computed tomography in partial carpal arthrodesis. AB - We carried out X-rays and computed tomography in 59 wrists in patients who had previous surgical intercarpal fusions. 1.2 mm thick axial images were obtained perpendicular to the axis of the joint. CT showed whether or not the carpal fusions were united. Compared with CT, plain radiography yielded a 25% false negative and 6% false positive rate. We conclude that CT is more useful than plain X-rays for evaluating partial carpal arthrodesis. PMID- 9752926 TI - Operative fluoroscopy in hand and upper limb surgery. One hundred cases. AB - We reviewed the use of a low radiation portable fluoroscopy unit in 100 patients. The most common indication was closed reduction of distal radial fractures. Fracture and joint stability were assessed on the real-time monitor and stored on videotape. Static images were stored on thermographic paper. Fluoroscopically guided joint injections and localization of implants, foreign bodies and bone tumours were performed. Fluoroscopy is a useful adjunct to arthroscopic assisted fracture reduction and other arthroscopic procedures such as distal ulnar resection. These new generation units produce superior resolution images, are easy to manoeuvre and do not require a radiographer. PMID- 9752927 TI - X-ray characteristics of wrists in calcium pyrophosphate crystal deposition disease. Is pseudogout a major cause of scapholunate advanced collapse? AB - Deposition of calcium pyrophosphate dihydrate (CPPD) crystals has been considered to be a cause of scapholunate advanced collapse (SLAC) wrist. The aim of this study was to look at X-ray changes in wrist joints affected by CPPD crystal deposition disease and to determine whether crystal deposition is a cause of SLAC wrist. A total of 150 wrists of 78 patients with CPPD crystal deposition disease were examined. In our population of Japanese patients with CPPD crystal deposition disease, the incidence of SLAC wrist was very low, and no case of Stage III SLAC wrist was found. We therefore conclude that SLAC wrist is not a radiographic characteristic of CPPD crystal deposition disease and that pyrophosphate crystal deposition cannot be a major cause of SLAC wrist. PMID- 9752928 TI - Thoracic outlet compression syndrome caused by a schwannoma of the C7 nerve root. AB - This is the first report of a schwannoma originating from the C7 nerve root causing thoracic outlet compression syndrome. The patient was a 30-year-old woman with a 3-year history of numbness on the radial side of the left hand, left arm tiredness, nocturnal pain in the left forearm and pain in the left elbow, shoulder and neck. Conservative treatment and previous operations, including carpal tunnel release and first rib resection, provided no relief. A left scalenectomy was performed. During the removal of the anterior scalene muscle, a mass approximately 3 cm long and 1.5 cm in diameter was noted under the anterior scalene muscle involving the C7 nerve root. The tumour was encapsulated and covered with attenuated and stretched nerve fascicles. It was completely excised without disturbing the nerve fascicles. The clinical impression was schwannoma, which was confirmed on pathological examination. PMID- 9752929 TI - Multiple schwannomas in the radial nerve. AB - We present a case of multiple schwannomas in the radial nerve. The occurrence of multiple schwannomas in a single major nerve is very rare. Magnetic resonance imaging was useful in detecting the tumours. As schwannomas may be multiple without clinical symptoms, we recommend MR imaging of the entire limb when schwannomas occur in a major nerve in the upper extremity. PMID- 9752930 TI - Avulsion fracture of the metacarpophalangeal joint of the finger. AB - Six patients with avulsion fractures of the metacarpophalangeal joints of the fingers are reported. Operation was performed in all cases. Judging from the operative findings, the radiological assessment of fragment shape is helpful in treatment. Surgery is recommended when the fragment is triangular or rectangular in shape because the fracture involves the articular surface. Conservative treatment is effective if the fragment is round because the articular surface of the joint is not involved in this type of fracture. The avulsed fragment often overlaps the metacarpal head and a collateral ligament injury is likely to be misdiagnosed. It is important to suspect this injury and assess the shape of the whole fragment for a good functional result. PMID- 9752931 TI - Late treatment of a displaced intraarticular metacarpal head fracture. PMID- 9752932 TI - Congenital palmar nail syndrome. AB - A rare case of congenital palmar nail syndrome is described. Nomenclature of this rare form of ectopic nail is discussed and a classification of the palmar nail deformity is offered. PMID- 9752933 TI - Extensive nodular sarcoidosis in the hand. AB - We report an atypical case of nodular sarcoidosis involving both hands. The pattern of extensive involvement of all digits with lesions extending into the pulp spaces has not been reported previously. The diagnosis of sarcoidosis should be considered even in patients presenting with clinically uncharacteristic manifestations. PMID- 9752934 TI - Upper limb Escherichia coli cellulitis in the immunocompromised. AB - The neutropenic state characteristic of acute lymphoblastic leukaemia (ALL) predisposes to infections involving Gram-negative bacilli. An Escherichia coli cellulitis originating in the first web space of the hand is described in a patient undergoing reinduction chemotherapy for ALL. Proximal extension of the infection progressed at a very rapid rate and required a forequarter amputation as a life saving measure. Due to the blunted inflammatory response in neutropenic patients, the need for close monitoring and quick intervention is stressed. PMID- 9752935 TI - Use of the Odstock wire as a simple method of evaluating multi-articular function in digits. PMID- 9752936 TI - Closed rupture of the flexor tendon caused by crystal deposition in the triangular fibrocartilage complex (TFCC) PMID- 9752937 TI - Diabetes and trigger finger. PMID- 9752938 TI - Preventive health pamphlets in the emergency department. AB - We conducted a prospective clinical trial to determine the effectiveness of an emergency department informational pamphlet in improving patients' compliance with recommendations that they receive Pap smears, mammograms, and a pneumococcal vaccination. Informational pamphlets were distributed to 1,000 consecutive patients who presented to a university-affiliated emergency department (ED). The pamphlet contained information stating the indications for obtaining routine Pap smears, mammograms, and a pneumococcal vaccination. Target individuals were women 18 years and older and men 65 years and older. Target patients were called approximately 2 months after their ED visits to obtain follow-up data. There were 464 target patients obtained from the 1,000 pamphlets distributed (409 female/55 male), and 68% (316) of the 464 were contacted by telephone for follow-up data. Significantly more women than men had read the pamphlet (62% vs. 8%). Of the women contacted (279), 31.9% (89) were not up to date (UTD) with Pap smears, and 11.2% (10) stated that they had scheduled an appointment for a Pap smear; 14.5% (11) of the women were not UTD with mammograms, and none had scheduled an appointment to receive care. Of the patients over age 65, 67% were not UTD with a pneumococcal vaccination, and no appointments were scheduled to obtain one. We conclude that a significant number of patients who present to this ED are in need of preventive health care. Emergency department informational pamphlets may have a role in improving Pap smear compliance. Women may be more likely then men to read informational pamphlets distributed in the ED. PMID- 9752939 TI - Emergency department visits for carbon monoxide poisoning in the Pacific Northwest. AB - This study was conducted to determine the annual number of emergency department (ED) visits and rate of hyperbaric oxygen (HBO2) treatment for carbon monoxide (CO) poisoning in Washington, Idaho, and Montana. All hospital emergency departments and hyperbaric treatment facilities in the region were surveyed by mail and telephone regarding their patient treatment experience for calendar year 1994. Results demonstrated that there were approximately 2.51 million total ED visits in 1994 in the three states studied. Among these, an estimated 1,325 individuals were seen with carbon monoxide poisoning (52.9 CO cases per 100,000 ED visits; 18.1 CO cases per 100,000 population). A total of 91 patients were treated with HBO2, yielding an HBO2 treatment rate of 6.9% of those evaluated in EDs. Extrapolating these figures to the US population suggests that the number of individuals seeking emergency medical care for CO poisoning is much greater than is commonly quoted. Even after correcting for the known increased rate of CO poisoning in the Pacific Northwest, the incidence of nonfatal poisoning appears to be significantly higher than may be appreciated from previous reports. PMID- 9752940 TI - Quantitative B-hCG levels less than 1000 mIU/mL in patients with ectopic pregnancy: pelvic ultrasound still useful. AB - The purpose of this study was to determine if pelvic ultrasound was useful in suggesting the diagnosis of ectopic pregnancy in patients with a quantitative B hCG level less than 1000 mIU/mL. We performed a retrospective review of all patients evaluated and diagnosed with ectopic pregnancy in the emergency departments of seven area hospitals during a ten month period. Sixty-four patients with a confirmed diagnosis of ectopic pregnancy, a pelvic ultrasound, and a quantitative B-hCG level were included in the study. Eighteen (28%) of these patients had a quantitative B-hCG less than 1000 mIU/mL. Sixteen of the eighteen patients (89%) with a B-hCG level less than 1000 mIU/mL had sonographic findings suggestive of ectopic pregnancy, such as fluid in the cul-de-sac, or a complex adnexal or cystic mass. Overall, 25% of all patients diagnosed with an ectopic pregnancy during this time period had a quantitative B-hCG level less than 1000 mIU/mL and an ultrasound suggestive for ectopic pregnancy. Pelvic ultrasound is useful as a screening tool in the initial evaluation of suspected ectopic pregnancy, even when the quantitative B-hCG level is below 1000 mIU/mL. PMID- 9752941 TI - The presentation of tetanus in an emergency department. AB - Traditionally, the literature has described a certain population as at risk for tetanus infection. We reviewed the demographics, clinical presentation, laboratory findings, management, and outcome of all patients who presented to our emergency department (ED) with tetanus in the last 10 years and compared our experience with this classic literature. We performed a retrospective case series review at a large, inner-city medical center; 11 cases of tetanus were identified from 1986 to 1997. Nine male and two female patients were identified with an average age of 45 years. All had an acute injury to the skin, and most (82%) reported having no history of recent immunization. The most common recorded symptoms were trismus and rigidity in the abdomen, neck, back, or extremities. There was only one misdiagnosis in the ED. Three patients died in the hospital, while the other eight were discharged either home or to a rehabilitation facility. In contrast to the classic literature, we found that tetanus in our inner city ED presented in recent immigrants, particularly younger men, over half of whom had received no childhood immunization. Laboratory results and cultures are of little diagnostic value, so timely recognition of the clinical presentation is important. PMID- 9752943 TI - Non-convulsive status epilepticus in a patient with hypocalcemia. AB - Non-convulsive status epilepticus (NCSE), a neurological emergency, is reported to account for approximately 25% of patients presenting in status epilepticus. Diagnosis of NCSE can be delayed or missed because of its often subtle presentation. Hypocalcemia has rarely been reported as a precipitator of NCSE and thus should be considered in the differential. We report the case of a 46-year old man with idiopathic hypoparathyroidism who presented in NCSE secondary to hypocalcemia. As in patients with convulsive status epilepticus, rapid diagnosis and treatment of patients in NCSE is critical to prevent permanent neurological damage. PMID- 9752942 TI - Elderly patients with closed head trauma after a fall: mechanisms and outcomes. AB - Falls in the elderly leading to closed head trauma represent a significant cause of morbidity and mortality in that population, but are not well-characterized. The purpose of this study was to determine the mechanism of fall, outcome, and additional risk factors in elderly patients who require cranial computed tomography (CT) scan after a fall. We conducted a retrospective case series of patients age 60 years and older with closed head trauma secondary to falling who underwent CT scan in the emergency department (ED). Data were gathered from ED and hospital records. The setting was an urban Level I trauma center. Our series consisted of 189 patients, of whom 31 (16%) had an abnormal head CT scan and four (2%) required neurosurgery. Cerebral contusions (38%) and subdural hematomas (33%) were the most common lesions seen on CT scan. Falls from standing (76%) were more common than falls on stairs (19%) or from height (5%), but the latter two were more likely to result in an abnormal CT scan (stairs 42%, height 40%). An abnormal neurologic examination was associated with a higher risk of the need for neurosurgery (risk ratio 11.5). We conclude that among elderly patients who fall and present to an ED with evidence of closed head trauma, a significant percentage will have abnormal CT scans but only a small minority will require neurosurgery. While falls from standing are more common, falls on stairs or from height are associated with a higher risk of having an abnormal CT scan. A focal neurologic examination is a strong predictor of the need for neurosurgical intervention. PMID- 9752944 TI - Central nervous system tuberculoma: a case report. AB - Pediatric cerebral tuberculoma is a disease rarely encountered in the United States. We report a case of central nervous system tuberculoma in a 6-month-old infant who presented to the emergency department with isolated right upper extremity paralysis. The discussion includes a brief review of central nervous system tuberculomas. PMID- 9752945 TI - Rapid reversal of life-threatening toluene-induced hypokalemia with hemodialysis. AB - A case of toluene-induced hypokalemia, respiratory failure, metabolic acidosis, and ventricular tachycardia is presented. Hemodialysis was associated with rapid improvement in our patient. PMID- 9752946 TI - Clonidine and sleep apnea syndrome interaction: antagonism with yohimbine. AB - A patient with sleep apnea syndrome, concurrently taking clonidine as an antihypertensive, presented with severe respiratory acidosis, hypotension, and associated central nervous system depression. Acidosis was improved by mechanical ventilation, and central nervous system (CNS) depression and hypotension were reversed with yohimbine. Clonidine may have an additive CNS depressive effect in sleep apnea syndrome and should be used with caution in such patients. Yohimbine's sympathetic-enhancing effects may be useful in clonidine toxic states. PMID- 9752947 TI - Flight paramedic scope of practice: current level and breadth. AB - This study's objective was to determine the current level and breadth of flight paramedic scope of practice. A six-item survey of lead flight paramedics in 158 air medical programs addressed five issues: 1) Certifications required above state certification; 2) Procedures included in scope of practice; 3) Medications flight paramedics are allowed to administer; 4) Requirements needed to expand scope of practice; and 5) Views on establishing a National Flight Paramedic Certification to alter their scope of practice. Eighty programs out of the 90 respondents (89%) stated that they utilize flight paramedics. Of these 80 programs that use flight paramedics, 76 programs (95%) require certification in ACLS, 65 (81%) in PALS, and 50 (63%) in BCLS. Paramedics are allowed to perform cricothyroidotomy in 68 programs (85%), pericardiocentesis in 24 (30%), and tube thoracostomy in 23 (29%). Medications approved for administration include streptokinase in 37 programs (46 %), r-TPA in 48 (60%), and succinylcholine in 50 (63%). In 61 programs (76%), the scope of practice is determined solely by the air medical director. Eighteen respondents (23%) believe that the development of a National Flight Paramedic Certification Program would alter their scope of practice. In conclusion, flight paramedic scope of practice varies enormously. Since most medical directors have the authority to alter flight paramedic scope of practice and few programs believe that a National Flight Paramedic Certification would alter their practice, medical directors should work directly with flight paramedics to expand their scope of practice. PMID- 9752948 TI - Postpericardiotomy syndrome following minimally invasive coronary artery bypass. AB - A 62-year-old woman presented to the Emergency Department (ED) with chest pain, cough, subjective fever and chills. Symptoms had begun on the previous evening, three days after minimally invasive coronary artery bypass surgery (MICAB). A presumptive diagnosis of postpericardiotomy syndrome (postcardiac injury syndrome) was made and the patient admitted. This new, minimally invasive surgery allows discharge on the second postoperative day. Emergency physicians should be aware of this procedure since probably there will be increasing performance of MICAB procedures and patients will present to the ED with postoperative complications. PMID- 9752949 TI - Entrance and exit gunshot wounds: incorrect terms for the emergency department? AB - Gunshot wounds (GSWs) pose significant medicolegal and forensic issues. Mistakes have been made regarding identification of gunshot wounds. We present a case of an atypical gunshot wound and review contact, near contact, intermediate, and distant wounds. There is an objective terminology to describe GSWs in the Emergency Department. PMID- 9752951 TI - Acute intermittent abdominal pain. PMID- 9752950 TI - The esophageal detector device can give false positives for tracheal intubation. AB - The esophageal detector device is a useful, inexpensive, portable device for the detection of inadvertent intubation of the esophagus. It has been extensively tested. It is very accurate with occasional false negatives for tracheal intubation but only two reported cases of false positives for tracheal intubation previously described. This article presents two further cases of false positives for tracheal intubation using the esophageal detector device and warns that although it is the more accurate of the cheap and portable devices available, it is not perfect. PMID- 9752952 TI - Bilateral edema of conjunctivae and eyelids due to massive subarachnoid hemorrhage and elevated intracranial pressure. PMID- 9752953 TI - Questioning thrombolytic use for cerebrovascular accidents. PMID- 9752954 TI - Predicting scar formation: from ritual practice (Langer's lines) to scientific discipline (static and dynamic skin tensions) PMID- 9752955 TI - Should we honor prehospital DNR orders in patients who attempt suicide? PMID- 9752956 TI - Emergency medicine resources on the Internet. AB - The Internet is a rapidly developing and useful tool for emergency physicians. A brief history of the development of the Internet and applications to Emergency Medicine are reviewed. A listing of many useful Internet resources is included. PMID- 9752957 TI - Is stapedectomy ever ethical? PMID- 9752958 TI - The efficacy of hyaluronic acid foam as a middle ear packing agent in experimental tympanoplasty. AB - The efficacy of hyaluronic acid (HA) foam in the prevention of middle ear adhesions and other structural abnormalities in guinea pigs undergoing experimental tympanoplasty was investigated. Postoperative changes in the middle ear were evaluated by light microscopy after 6 weeks. The presence of adhesions, diminution of airspace, new bone formation, tympanic membrane (TM) formation, and mucosal inflammation was characterized by an objective grading system. Results were compared to absorbable gelatin sponge (AGS) and a control group (no middle ear packing). The control group showed the best average scores for all parameters tested except for adhesion formation. However, these results were statistically significant only when compared with those of the AGS group for airspace preservation, new bone formation, and TM formation. Although the HA foam group showed better average results than did the AGS group for all parameters tested, none were statistically significant. Although HA foam appears to be a promising middle ear packing agent, further experimental trials are warranted before any firm conclusions may be made. PMID- 9752959 TI - Mastoidectomy elimination: obliterate, reconstruct, or ablate? AB - OBJECTIVE: This study aimed to evaluate a chronology of techniques used to manage troublesome open mastoid cavities, with emphasis on the selection of the mastoidectomy elimination technique most appropriate to the case at hand. STUDY DESIGN: The study design was a retrospective review of techniques used in 465 consecutive elimination cases. SETTING: The study was conducted at a single surgeon's private otologic practice. PATIENTS: Patients with mastoidectomy elimination who were treated from 1974-1996, including 55 patients with obliterations (cavity fill-in), 372 patients with reconstructions (canal wall repair), and 38 patients with ablations (external canal closure), requiring 823 procedures, were examined. MAIN OUTCOME MEASURES: Clinical success and complication rates of the techniques studied were measured. RESULTS: Optimal outcomes (89% successful) were recorded from hydroxylapatite reconstruction cases managed with canal revascularization (middle temporal flap) and cholesteatoma prevention (staging and composite grafts). CONCLUSIONS: Obliteration is recommended only over noncholesteatomatous sites because of the risk of residual disease and the difficulty re-exploring these cases. Ablation is effective in selected, severely damaged cases. All ablation cases remained disease-free after surgery. Reconstruction is the preferred method when hearing restoration is required. Canal wall reconstruction required modifications to avoid complications from poor tissue vitality and cholesteatoma. These are outlined and discussed below. PMID- 9752960 TI - Cranial anatomy and otitis media: a cadaver study. AB - BACKGROUND: The eustachian tube is regarded as an etiologic factor for otitis media. Although anatomic cranial differences also are suggested as a factor, few scientifically rigorous studies of these differences have been reported. MATERIALS: Thirty-five adult cadaveric crania were examined. METHODS: Multiple (32) linear and angular cranial measurements were performed. For evidence of prior otitis media, there were two indicators: small mastoid pneumatization seen radiographically and abnormal tympanic membranes at photographic tympanoscopy. Each measurement and each categorization were done twice, independently. The average of the two measurements was used for each comparison. Only consistent categorizations were used for comparison. RESULTS: Relatively short eustachian tubes were found to associate with both indicators of childhood otitis: r=0.39, p < 0.05. A relatively short distance from midsella turcica to staphylion, and short distance between the ears, also were associated with otitis. No angular relation of either the bony or cartilaginous eustachian tube correlated with the otitis indicators. Bilateral symmetry of pneumatization and tympanoscopic categorization, and of the various linear and angular measurements, was apparent. CONCLUSIONS: The association of otitis media with some cranial base anatomic differences is endorsed. Comparatively long eustachian tubes, long distance from midsella to staphylion, and large interear length correlate with indicators of healthy middle ears. PMID- 9752961 TI - Results of cavity reconstruction with hydroxyapatite implants after 15 years. AB - OBJECTIVE: The results of the first cohort of 60 cavity reconstructions with hydroxyapatite with a minimum follow-up period of 15 years were studied. STUDY DESIGN: The study design was a retrospective study. PATIENTS: A total of 60 patients had a follow-up period of >15 years. In four patients, not all data were available. Therefore, 56 patients were included in the study. They had a combination of cavity problems and hearing loss. INTERVENTION: The ear canal was reconstructed with a canal wall prosthesis of porous hydroxyapatite. The ossicular chain was reconstructed with an incus or incus-stapes prosthesis of dense hydroxyapatite. RESULTS: After 15 years, 42 patients (75%) had an intact reconstructed ear canal. The main problem for failure was the recurrent purulent middle ear infection and not cholesteatoma. The histology of the retrieved canal wall showed a good remodeling in living bone tissue. After 15 years, 34 patients had a normal ear canal and an ossicular chain. Of these patients, 7.05% had an air-bone gap closure within 20 dB. CONCLUSIONS: Long-term results of cavity reconstruction with hydroxyapatite are possible. The main problem is recurrent mucosal disease of the new middle ear-mastoid cleft. PMID- 9752962 TI - Titanium as an ossicular replacement material: results after 336 days of implantation in the rabbit. AB - OBJECTIVE: Titanium in other parts of the body, well known for its biocompatibility, was examined in an animal model for its use as an ossicular replacement material. STUDY DESIGN: The biocompatibility of titanium was studied in the middle ear of rabbits using light and scanning electron microscopy. Titanium pins were placed as middle ear prostheses or as free implants and were examined after 28, 84, 168, and 336 days. RESULTS: After 28 days, the prostheses were covered by regular mucosa. The free implants took up to 336 days to be totally epithelialized. There were no inflammatory cells observed on the surface of the material nor were unusual amounts of fibrous tissue seen. In addition, the titanium material exhibited an affinity toward bone. CONCLUSIONS: The results of this animal experiment indicate that titanium is a useful material for ossicular replacement prostheses. PMID- 9752963 TI - A randomized, blinded study of canal wall up versus canal wall down mastoidectomy determining the differences in viewing middle ear anatomy and pathology. AB - HYPOTHESIS: Canal wall down and intact canal wall tympanomastoidectomy represent two surgical approaches to middle ear pathology. The authors hypothesize that there is a difference in the ability to view structures in the middle ear between these two methods. BACKGROUND: Depending on the individual, many surgeons have used the two different techniques of intact canal wall and canal wall down tympanomastoidectomy for approaching the middle ear. However, opinions conflict as to which approach provides the best visualization of different locations in the middle ear. This study prospectively evaluated temporal bones to determine the differences in visualizing structures of the middle ear using these two approaches. METHODS: Twelve temporal bones underwent a standardized canal wall down tympanomastoidectomy using a reversible canal wall down technique. All bones were viewed in two dissections: intact canal wall and canal wall down preparations. Four points previously had been marked on each temporal bone in randomly assigned colors. These points include the sinus tympani, posterior crus of stapes, lateral epitympanum, and the Eustachian tube orifice. An observer blinded to the purpose of the study, color, and number of locations recorded the color and location of marks observed within the temporal bones. Randomized bones of two separate settings were viewed such that each bone was viewed in both the canal wall down and the intact canal wall preparations. RESULTS: A significant difference was noted in the ability to observe middle ear pathology between the intact canal wall versus canal wall down tympanomastoidectomy, with the latter showing superiority (p < 0.001). Of the four subsites, the sinus tympani, posterior crus of stapes, and lateral epitympanum were observed more frequently with the canal wall down. There was no significant difference in the ability to observe the Eustachian tube orifice between the two techniques. CONCLUSIONS: Statistical analysis shows good reproducibility and randomization of this study. The canal wall down tympanomastoidectomy allowed for superior viewing of the three locations, sinus tympanic, posterior crus of stapes, and lateral at the tympanum, as they were marked in the study. This study shows the potential for improved visualization via the canal wall down tympanomastoidectomy. A significant amount of literature written by individuals and otology group practices is available retrospectively comparing the advantages and disadvantages of intact canal wall versus canal wall down mastoidectomy procedures for approaching middle ear pathology. In the interest of objectively evaluating the differences between these two approaches, we have studied temporal bones in a prospective randomized, blinded study comparing the two. Twelve bones were used and observed twice, once in each of 2 sessions. All bones were viewed in two dissections: intact canal wall and canal wall down mastoidectomy. Four points were marked on each temporal bone in three different colors applied in a randomized order to eliminate observer expectation. The four points marked include sinus tympani, posterior crus of the stapes footplate, lateral epitympanum, and Eustachian tube orifice. Both intact canal wall and canal wall down bones were provided randomly to the observer at each viewing session. Before the observer was allowed to see the dissections, those requiring replacement of the canal for the first session of the study had this done in a method using native posterior bony canal. Temporal bones were presented to an expert otologist in a randomized fashion with each temporal bone being placed in a temporal bone bowl holder and specialized framework, allowing for rotation and repositioning approximating the experience in an operating room setting. For each temporal bone, the observer filled in a questionnaire describing his or her observations by denoting both location and color of marks observed. (ABSTRACT TRUNCATED) PMID- 9752964 TI - What kind of patients are suitable for evaluating the therapeutic effect of sudden deafness? AB - OBJECTIVE: This study aimed to find out appropriate patients to evaluate the therapeutic effect of sudden deafness. STUDY DESIGN: The study design was a retrospective case review. SETTING: The study was performed at a university hospital. PATIENTS: A series of 443 patients with idiopathic sudden sensorineural hearing loss who visited the authors within 1 week from the onset participated. INTERVENTIONS: Relationship between interval from the onset and treatment prognosis was investigated. RESULTS: The significant difference in prognosis related to the days from the onset to the initial visit was noted only in the patients with the initial hearing level from 50-65 dB. The prognosis of the patients who visited the authors on the next day from the onset was excellent. Whereas, in the patients with initial hearing level worse than 70 dB, no significant difference in prognosis was noted among the days of visit. CONCLUSIONS: The patients with the initial hearing level worse than 70 dB and who visited the authors within 8 days from the onset were appropriate for candidates in evaluating the therapeutic effect of sudden deafness. PMID- 9752965 TI - Magnetic resonance imaging compatibility testing of the Clarion 1.2 cochlear implant. AB - OBJECTIVE: This study aimed to investigate the compatibility of the Clarion 1.2 magnet-containing cochlear implant with a 1.5-tesla (T) and 0.3-T magnetic resonance imager. BACKGROUND: Cochlear implants restore functional hearing to patients with sensorineural deafness. With the rapidly increasing number of patients with cochlear implants, there is a need to investigate the implant's magnetic resonance imaging (MRI) compatibility. METHODS: The authors tested the potential torque and force on the metallic components of the implant, heating of the implant and surrounding tissue, unintentional output, implant damage, and image distortion. Tests were performed in both a 1.5-T and 0.3-T MRI. RESULTS: The torque experienced by the implant in the 1.5-T MRI (0.19 nm) was large enough that it could potentially cause implant movement in some patients. An acceptable amount of torque (0.04 nm) was found in the 0.3-T MRI. Image distortion occurred in the area directly around the implant with a radius of up to 60 mm in the 1.5-T MRI and 100 mm in the 0.3-T MRI. In both MRI units, there was no detectable temperature increase or unintentional output. There was no implant damage except that with worst-case conditions, the internal magnet was demagnetized by 78.5% with the 1.5-T unit and 3.36% with the 0.3-T unit. CONCLUSIONS: The authors recommend patients with cochlear implants avoid imaging in a 1.5-T MRI. The results suggest that the 0.3-T MRI poses little or no risks to patients with cochlear implants. PMID- 9752966 TI - Cochlear implants for adults obtaining marginal benefit from acoustic amplification: a European study. AB - OBJECTIVE: This study aimed to examine the application of a speech recognition score of 30% on open-set word materials as the upper limit for preoperative performance in determining cochlear implant (CI) candidacy for European non English-speaking hearing-impaired persons. This study also aimed to determine the effect of implantation on residual pure-tone hearing thresholds and to determine the incidence and benefit of a contralateral hearing aid postimplant. STUDY DESIGN: The single-subject design study, involving 20 postlinguistically deafened subjects, compares preoperative performance with hearing aids to postoperative performance with a CI at 6 months after surgery. Subjects were implanted with either the Nucleus Mini 22 or the Nucleus 24 CI systems implementing the MPEAK and SPEAK coding strategies. Fourteen subjects meeting the selection criteria were accrued consecutively specifically for inclusion in the study, whereas the remaining 6 retrospectively implanted subjects were identified for inclusion via patient records. PATIENTS: The investigation included 8 clinics over 3 countries (France, Germany, and Spain) and involved 20 postlinguistically deafened subjects who obtained marginal benefit from acoustic amplification before surgery. Nineteen subjects were older than 18 years of age with 1 subject being 14 years old included in the data report as well. MAIN OUTCOME MEASURES: Open-set speech recognition was evaluated before and after surgery using recorded word lists and sentence lists in the subject's native language to determine benefit from the treatment. Baseline audiograms were obtained before surgery for frequencies of 0.25-8.0 kHz for both ears and compared to pure-tone hearing thresholds measured at 1 month after surgery to determine the effect of the implantation on residual hearing. Additionally, a questionnaire was administered to determine the incidence and benefit of continued hearing aid use in the contralateral ear postimplant. RESULTS: Nineteen of the 20 study subjects displayed a significant benefit after surgery at 6 months after switch-on for open-set speech recognition. The remaining subject displayed no significant change in performance on objective testing. The implantation resulted in a significant downward shift in hearing thresholds for the implant ear in the majority of subjects. However, 50% of subjects displayed conservation of some residual hearing. For the majority of subjects, hearing aid use in the contralateral ear was discontinued because of lack of perceived benefit after surgery. CONCLUSIONS: The Nucleus Multichannel CI provides a significant benefit for postlinguistically deafened adults who display marginal benefit from acoustic amplification. Therefore, in French-, German-, and Spanish-speaking clinics, a speech recognition score of 30% on open-set word materials is considered an appropriate upper limit for preoperative performance in determining CI candidacy. In view of a significant downward shift in pure-tone thresholds in the implant ear for the majority of subjects, in cases of asymmetry, it is recommended that the poorer ear be implanted. After surgery, the majority of subjects did not perceive an added benefit from continued use of their contralateral hearing aid. PMID- 9752967 TI - The influence of the concentration of volatile anesthetics on the stapedius reflex determined intraoperatively during cochlear implantation in children. AB - OBJECTIVE: The goal of this study was, first, to determine the effect of the concentrations of volatile anesthetics (halothane and isoflurane) on the intraoperative elicited electrical stapedius reflex threshold (ESRT) and, second, to evaluate the relation between the ESRTs and postoperative C-levels. STUDY DESIGN: This was a prospective clinical study in a single subject design. SETTING: The study was conducted at University Hospital Nijmegen, which is a tertiary care and cochlear implant center in The Netherlands. PATIENTS: The study population comprised 13 deaf children (6 males and 7 females) undergoing cochlear implantation. RESULTS: In most of the children, increasing the concentration of volatile anesthetic agent resulted in higher stapedius reflex threshold. After correction for this effect, a good relation was found between ESRT and C-level in all children, except for one. CONCLUSIONS: The outcome of this study supports the determination of intraoperative ESRTs for programming of the speech processor postsurgery. PMID- 9752968 TI - Association of COL1A1 and otosclerosis: evidence for a shared genetic etiology with mild osteogenesis imperfecta. AB - HYPOTHESIS: Otosclerosis is related to mild osteogenesis imperfecta with genetic defects in type I collagen. BACKGROUND: Otosclerosis is a common bone disease of the human otic capsule that has an underlying hereditary predisposition. The histopathology and clinical manifestations are strikingly similar to the milder forms of osteogenesis imperfecta in which mutations of type I collagen genes have been established as the underlying cause. METHODS: The authors investigated the genetic basis of otosclerosis by conducting an association study using polymorphic DNA markers from patients with clinical otosclerosis and random control subjects. RESULTS: This study showed a significant association between clinical otosclerosis and the type I collagen COL1A1 gene using three different polymorphic markers within the gene. CONCLUSIONS: Some cases of clinical otosclerosis may be related to mutations within the COL1A1 gene that are similar to those found in mild forms of osteogenesis imperfecta and result in null expression of the mutant allele. PMID- 9752969 TI - Auditory reaction times in patients with chronic tinnitus with normal hearing. AB - HYPOTHESIS: This study aimed to compare reaction times (RTs) to auditory stimuli of two groups of normal-hearing subjects differing only in terms of tinnitus sensation. BACKGROUND: The RTs to auditory stimuli as a psychophysical measurement for threshold and suprathreshold hearing are said to provide a behavioral clue to some aspects of neural processing in the auditory system. METHODS: To explore how patients with tinnitus perceive the intensity of threshold and suprathreshold sound stimuli, RTs were obtained from normal-hearing subjects with tinnitus (experimental group, N=15) and from normal-hearing subjects without tinnitus (control group, N=15) by means of exposure to two different sets of frequencies: the tinnitus frequencies and the nontinnitus frequency of 1,000 Hz. RESULTS: There were significant differences in RTs in the experimental group and in the control group not only for the tinnitus frequencies, but also for the nontinnitus frequency of 1,000 Hz. The experimental group had shorter RTs than did the control group at sensation levels (SLs) near the threshold, with no significant differences between groups at sound stimuli in the suprathreshold intensity range. CONCLUSIONS: It is assumed that the above mentioned reduction in RTs shows a dysfunction of cochlear mechanisms contributing to tinnitus. Conversely, tinnitus also can be considered as an additional auditory input leading to shorter RTs at SLs near the threshold. The current study suggests that the reaction time procedure to auditory stimuli offers complementary information on tinnitus sensation and might be a valuable method in demonstrating general differences and tendencies that have been neglected so far. Analysis for the mechanisms of tinnitus sensation allows for the possibility of facilitating the process of tinnitus habituation and, ultimately, the relief from it. PMID- 9752970 TI - Evaluation of posturography in the detection of malingering subjects. AB - OBJECTIVE: This study aimed to test the performance of proposed methods for detecting malingering subjects on computerized dynamic posturography using one subject group in three situations (normal, malingering, vestibular weakness). STUDY DESIGN: The study design was a prospective, blinded study. SETTING: The study was conducted at a university hospital. PATIENTS: Volunteer subjects aged 20-59 years of age participated. INTERVENTIONS: Computerized dynamic posturography was performed under three situations: best effort, faking vestibular weakness, and transient induced vestibular weakness with bilateral simultaneous caloric irrigation. MAIN OUTCOME MEASURES: Measured was identification of situation (normal, malingering, induced vestibular weakness) by each of three detection methods: blinded clinical scoring, a set of formulae, and a set of variables (the latter two methods proposed previously by other investigators). RESULTS: Each method performed well. In three-way discrimination, the formulae and clinical scoring each correctly identified approximately 75% of trials. In two-way discrimination (malingering vs. induced vestibular weakness), the best combination of variables slightly outperformed clinical scoring (0.93 vs. 0.88 ROC [receiver operating characteristic] curve area). CONCLUSIONS: Computerized dynamic posturography can distinguish malingering in normal subjects from trials performed with best effort or after binaural simultaneous caloric irrigation. The accuracy of blinded clinical scoring was comparable to that of two objective detection methods. PMID- 9752971 TI - Meniere's disease: evidence of an immune process. AB - OBJECTIVE: This study aimed to present further evidence of specific antibodies reactive against bovine inner ear proteins found in patients with Meniere's disease. BACKGROUND: There are a growing number of experimental and clinical features of Meniere's disease that indicate an immune association. Circulating antibodies directed against inner ear proteins have been detected in patients with bilateral progressive hearing loss and Meniere's disease. METHODS: A total of 36 patients with classical Meniere's disease were studied noting the presence of active or inactive, bilateral or unilateral disease. Bovine membranous labyrinth was used as inner ear extract and separated to molecular weights using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Western blotting included the addition of serum buffer (casein, Tween, and bovine serum albumins) and the use of alkaline phosphatase labeled anti-human immunoglobulin antibodies. RESULTS: The findings represent strong evidence of antibodies reactive to inner ear proteins. The incidence of these antibodies was correlated significantly with disease activity. CONCLUSIONS: The findings are further evidence of an immune process that is involved in Meniere's disease. PMID- 9752972 TI - A dual-task study of interference between mental activity and control of balance. AB - OBJECTIVE: This study aimed to examine interference between mental activity and control of balance. STUDY DESIGN: In a mixed design, dual-task study, the performance of patients and healthy control subjects was compared on computerized dynamic posturography, on a visuospatial mental task, and when performing the mental task while balancing. SETTING: The study was performed at a tertiary referral outpatient neuro-otology clinic. PATIENTS AND SUBJECTS: The patient group comprised 24 patients seen consecutively at the clinic because of vertigo and dizziness. The control group consisted of 24 subjects with no complaint or medical history of dizziness or balance disorder, matched with the patients for age and gender. MAIN OUTCOME MEASURES: Performance on a visuospatial mental task and on the computerized dynamic posturography test (conditions 4 and 5) was measured. RESULTS: Balancing on the posturography test resulted in a deterioration in performance on the mental task for both patients and control subjects. The effect was more marked when subjects had their eyes closed. Results on the balance test showed that normal subjects and patients with normal balance also swayed more when performing the mental task, whereas patients who had failed the posturography test swayed less when performing the mental task. CONCLUSIONS: These results show that mental performance deteriorates when performing a demanding balance task. In addition, in both normal subjects and patients, balance also may be affected by mental activity in complex and varied ways that merit further investigation. PMID- 9752973 TI - Hearing preservation in neurofibromatosis type 2. AB - OBJECTIVE: The aim of the study was to provide a clinical review of the middle fossa approach for hearing preservation in patients with neurofibromatosis type 2 (NF2). STUDY DESIGN: The study design was a prospective case review. SETTING: The study was conducted at a private practice tertiary neurotologic referral center. PATIENTS: Eighteen patients diagnosed with NF2 underwent 23 middle fossa procedures between 1992 and 1996 for removal of an acoustic neuroma. The nine males and nine females ranged in age from 11-73 years with a mean age of 28 years. Tumor size ranged from 0.5-2.5 cm with a mean of 1.1 cm. MAIN OUTCOME MEASURES: House-Brackmann facial nerve grade was measured. In addition, hearing level was classified both by the American Academy of Otolaryngology-Head and Neck Surgery criteria for reporting results of hearing preservation surgery and by comparison with preoperative level (15 dB/15%). RESULTS: Measurable hearing was preserved in 65%, 48% within 15 dB of preoperative pure-tone average (PTA) and within 15% of preoperative speech discrimination. Bilateral hearing preservation occurred in five patients. Excellent facial nerve function (House-Brackmann grades I-II) was obtained in all patients with normal preoperative facial nerve function. CONCLUSIONS: In this series of patients with NF2, outcomes in hearing and preservation of preoperative facial nerve function are similar to results seen in patients suffering a sporadic unilateral acoustic neuroma. However, early intervention is crucial in obtaining such favorable outcomes. PMID- 9752974 TI - Quality of hearing preservation in acoustic neuroma surgery. AB - OBJECTIVE: This study aimed to investigate the factors affecting the quality of postoperative hearing in acoustic neuroma. STUDY DESIGN: The study was designed as a retrospective case review. SETTING: The study was performed at the Department of Otolaryngology, School of Medicine, Keio University, Tokyo, Japan. PATIENTS: The subjects were 94 patients with unilateral acoustic neuroma. INTERVENTION: Hearing preservation surgery was performed in the subjects via the extended cranial fossa approach or the middle cranial fossa approach. MAIN OUTCOME MEASURES: The outcome measures included patient's age and gender, hearing level, speech discrimination score, tumor size, and surgical approach. The relationship between the qualities of preoperative and postoperative hearing and the long-term prognosis of preserved hearing also was investigated. RESULTS: In 94 subjects, there were 47 patients whose hearing was preserved (HP group) and 47 patients whose hearing was not preserved (non-HP group). Overall, hearing preservation rate was 50%. There were no significant differences in age, gender, and tumor size between the two groups. The hearing preservation rate was significantly higher in patients with an intracanalicular tumor than that with a larger tumor. The better the preoperative quality of hearing was, the higher the postoperative one. Although the preserved hearing deteriorated after surgery in 4 patients, no significant hearing deterioration was observed in the other 43 patients. CONCLUSIONS: The results of this study indicated that the diagnosis for acoustic neuroma in the early stage with serviceable hearing is the most important to improve the quality of postoperative hearing. PMID- 9752975 TI - Hearing restoration in posterior fossa tumors. AB - OBJECTIVE: This study aimed to assess the results of hearing restoration with a cochlear or a brainstem implant in posterior fossa tumors. PATIENTS: Six patients were selected. Two patients with an acoustic neuroma in the only-hearing ear (cases 1 and 2), one patient with a posterior fossa meningioma (case 3), one patient with bilateral facial neuroma (case 4), and two patients with bilateral acoustic neuroma (cases 5 and 6) participated. INTERVENTION: In cases 1 and 2, the patients had a cochlear implant inserted on the only-hearing ear opposite the acoustic neuroma. In case 3, the patient presented with total deafness on the left side and a 10-mm meningioma on the right side. A cochlear implantation was performed after removal of the meningioma on the right side. In case 4, the patient was operated on on both sides with bilateral postoperative deafness. A cochlear implantation was performed on the better hearing ear. In cases 5 and 6, patients underwent an auditory brainstem implantation after the exeresis of the second tumor. RESULTS: Promontory test results were positive for patients 1, 2, 3, and 4. After implantation, patients 1, 2, 3, and 4 scored 98%, 13%, 70%, and 30%, respectively, in open-set sentence recognition tests, whereas patients 5 and 6 scored 0% and 20%, respectively. CONCLUSIONS: In case of nonfunctional cochlear nerve, in acoustic neuroma, either bilateral and in the only-hearing ear, promontory test should be performed. If positive results, a cochlear implantation should be performed, because successful results could be expected. Overall results of cochlear implantation on speech discrimination are better than those obtained with a brainstem implant. PMID- 9752976 TI - Immunoblotting analysis of schwannomin/merlin in human schwannomas. AB - HYPOTHESIS: Absent or reduced expression of schwannomin/merlin is associated with tumorigenesis of sporadic schwannomas. BACKGROUND: The neurofibromatosis type 2 (NF2) gene frequently is mutated in sporadic vestibular schwannomas. The protein product of the NF2 gene is called schwannomin or merlin. Little is known about the mutated forms of schwannomin/merlin present in schwannomas. METHODS: To investigate further the role of schwannomin/merlin in schwannoma tumorigenesis, immunoblotting experiments were performed. Antischwannomin/merlin-specific antibody that recognizes amino terminus of the protein was used to determine the expression levels of schwannomin/merlin in 16 sporadic vestibular schwannomas, 1 NF2-related vestibular schwannoma, and 5 spinal schwannomas. RESULTS: The antibody detects a protein of approximately 66 kDa in the Triton X-100-insoluble fraction of tumors. The expression of schwannomin/merlin was severely reduced, <35% of control, in 11 (50%) of 22 sporadic schwannomas and in 1 NF2-related vestibular schwannoma. The intensity of 66-kDa schwannomin/merlin band was moderately reduced, from 35-60%, in 7 (32%) of 22 schwannomas compared to the expression levels found in the human brain. Truncated forms of schwannomin/merlin were identified in three tumors with moderately reduced schwannomin/merlin. CONCLUSIONS: These results provide new evidence that inactivation of schwannomin/merlin is an important factor in tumorigenesis of sporadic schwannomas. PMID- 9752977 TI - Hair cell formation in cultures of dissociated cells from the vestibular sensory epithelium of the bullfrog. AB - HYPOTHESIS: Bullfrog vestibular hair cells are capable of regenerating in vitro. BACKGROUND: Recent studies have established that sensory organs in the inner ear of vertebrates continue to produce hair cells after birth. However, the mechanisms responsible for the regulation of this process are not well understood. The current study reports the development of a novel method for the culture of dispersed cells from the bullfrog inner ear. METHODS: New hair cell formation in this in vitro preparation was shown by sequential photomicroscopy. Studies with the selective marker for mitotic activity 5-bromo-2-deoxyuridine (BrdU) were done to estimate the level of cell proliferation and to quantify postmitotic hair cell formation. Finally, confirmation of cell type was obtained by scanning electron microscopy and by the use of specific markers for hair cells. RESULTS: Once the optimal culture conditions were established in the initial experiments, the formation of new hair cells was directly visualized in all unstained live cultures and fixed preparations without exception. Asymmetric division of progenitor cells, with subsequent differentiation of one of the daughter cells into new hair cells, also was documented by photomicroscopy. Approximately 12% of the cells were labeled with BrdU, of which 6% were hair cells, showing that new hair cell formation was subsequent to mitotic division in vitro. The identity of newly formed hair cells was verified as follows: 1) morphologically by scanning electron microscopy; 2) by positive labeling with phalloidin-rhodamine, a marker for actin; and 3) by positive calmodulin immunocytochemistry. CONCLUSIONS: This study reports the development of an in vitro culture preparation in which undifferentiated epithelial cells proliferate to become new hair cells. Evidence is provided of division of hair cell progenitors and subsequent differentiation of the daughter cells as one of the mechanisms involved in new hair cell formation in the culture preparation. This newly developed cell culture technique provides a powerful tool for further study of the process of hair cell formation in the vestibular end organ. PMID- 9752978 TI - Congenital inner ear malformation. PMID- 9752979 TI - Teratogenic hearing loss: a clinical perspective. AB - OBJECTIVE: The Joint Committee on Infant Hearing recently published a series of indicators that place a neonate at risk for hearing loss. Included among these risk factors are environmental teratogens capable of negatively impacting the developing auditory system. This article serves to revisit and update potential environmental teratogens. The characteristic clinical presentation with audiometric, electrophysiologic, and temporal bone findings as well as available treatment options are discussed. DATA SOURCES: A Medline search encompassing the latter half of this century was undertaken to review pertinent literature regarding infectious, chemical, physical, and maternal teratogens and their impact on hearing impairment. CONCLUSIONS: Prevention and early recognition of environmental teratogenic exposure play a critical role in the reduction of childhood hearing loss and deafness. The importance of longitudinal follow-up in these patients is stressed. PMID- 9752980 TI - Indications for the surgical repair of unilateral aural atresia in children. AB - OBJECTIVE: This study aimed to clarify the indications for elective surgical repair of unilateral aural atresia in children and to review the rates of successful repair in the literature. DATA SOURCES: A search of the published English language literature, 1966-1997, was conducted using the key words aural atresia. STUDY SELECTION: Articles were selected on the basis of their inclusion of the authors' indications for surgical repair of aural atresia or the inclusion of surgical series that showed outcomes. DATA EXTRACTION: Articles reviewed in the review had to either provide specific guidelines for surgical repair of unilateral aural atresia or provide postoperative pure-tone averages, air-bone gaps, or speech reception thresholds. DATA SYNTHESIS: The authors compiled the relevant data into summary tables and extracted conclusions from these data. CONCLUSIONS: Elective surgical repair of unilateral aural atresia should only be attempted in children who meet specific anatomic criteria that predict that they are the most likely to benefit from the results of surgery. Otherwise, repair should be delayed until the age at which the patient can make an informed decision, knowing the risks, benefits, and consequences of this difficult surgery. Parents and surgeons must have a realistic expectation of the surgical results and the practiced benefit to be expected with a normal, contralateral ear. PMID- 9752981 TI - The past, present, and future of the prevention of lung cancer. AB - The article relates details of the history of research into the causal association of cigarette smoking and lung cancer on the basis of multidisciplinary studies that have explored the epidemiology, biology, chemistry, and biochemistry of tobacco carcinogenesis and research in behavioral sciences and health education that has sought to address one of our nation's foremost public health problems. Recalling past and present challenges and achievements in all of these areas, the author then outlines his vision for addressing this health problem in the future. This is laid out for various segments of the research community and for society as a whole, i.e., Cancer Centers and hospitals, epidemiologists, laboratory scientists, legislators, educators and behavioral scientists, and the media. It is proposed that for the current policy initiatives in tobacco-related cancer control to succeed, there needs to be a focus on preventing the initiation of tobacco use among children and adolescents. All segments of society can help to achieve this goal. In the nation's research planning, there needs to be a proper balance between basic and applied research, including research on and application of preventive principles, because cancer need not be an inevitable consequence of aging but is largely preventable. PMID- 9752982 TI - Risk of esophageal and gastric adenocarcinomas in relation to use of calcium channel blockers, asthma drugs, and other medications that promote gastroesophageal reflux. AB - Incidence of adenocarcinomas of the esophagus and gastric cardia has risen dramatically over the past 2 decades in the U. S., for reasons that are not yet clear. A number of common medications (e.g., calcium channel blockers, tricyclic antidepressants, and certain asthma medications) promote gastroesophageal reflux by relaxing the lower esophageal sphincter (LES). Reflux is thought to increase cancer risk by promoting cellular proliferation, and by exposing the esophageal epithelium to potentially genotoxic gastric and intestinal contents. Recent studies have suggested that calcium channel blockers may also increase cancer risk by inhibiting apoptosis. Using personal interview data from a multicenter, population-based case-control study conducted between 1993 and 1995 in three areas of the U. S., we evaluated whether the use of LES-relaxing drugs was associated with increased risk of adenocarcinomas of the esophagus and gastric cardia. Cases of esophageal adenocarcinoma (n = 293) and gastric cardia adenocarcinoma (n = 261) were compared with general population controls (n = 695). Information on additional case groups of esophageal squamous cell carcinoma (n = 221) and noncardia gastric cancer (n = 368) were also available for comparison. Overall, 27.4% of controls had used one or more of these drugs for at least 6 months, compared with 30.2% of esophageal adenocarcinoma and 23.8% of gastric cardia adenocarcinoma cases. The adjusted odds ratios (ORs) for ever use were 1.0 [95% confidence interval (CI) = 0.7-1.5] and 0.8 (95% CI = 0.5-1.1), respectively. There was little evidence of increasing risk with increasing duration of use of all LES-relaxing drugs together. We found an increased risk of esophageal adenocarcinoma among persons reporting use of asthma drugs containing theophylline (OR = 2.5; 95% CI = 1.1-5.6) or beta agonists (OR = 1.7; 95% CI = 0.8-3.8). Risks were higher among long-term users (>5 years) of these drugs (OR = 3.1; 95% CI = 0.9-10.3 and OR = 2.3; 95% CI = 0.8-7.0, respectively). In contrast, there was no evidence that the use of calcium channel blockers or other specific groups of drugs increased the risk of any of the cancers studied. These results provide reassuring evidence that the increases in incidence of adenocarcinomas of the esophagus and gastric cardia are not likely to be related to the use of LES-relaxing drugs as a group, or calcium channel blockers in particular, but they do suggest that persons treated for long-standing asthma may be at increased risk of esophageal adenocarcinoma. PMID- 9752983 TI - Effects of milk and milk products on rectal mucosal cell proliferation in humans. AB - Intake of dairy products and major dairy constituents (e.g., calcium) has been proposed to reduce the risk of colorectal cancer, although epidemiological studies have yielded inconclusive results. We conducted a randomized cross-over trial to test the effects of high- and low-dairy consumption diets on rectal mucosal proliferation, a possible intermediary marker for large bowel cancer. From a gastroenterology clinic at an academic medical center, we recruited 40 patients, ages 25-79 years, who had either a history of a large bowel adenoma or a first-degree relative with large bowel cancer. Participants completed a baseline questionnaire covering demographic characteristics, health history, and habits and a food frequency questionnaire. They were randomized to a 12-week diet of either high dairy intake (six dairy servings/day) or low dairy intake (<0.5 serving of dairy products/day), with an intervening 12-week washout period in which they were asked to resume their usual diet before crossing over to the alternate study diet for the last 12-week period of the study. Adherence to the study diets was monitored by a daily dairy intake checklist and periodic, unscheduled 24-h dietary recalls. Biopsies of the rectal mucosa were obtained at the beginning and end of each intervention phase. Two assays of rectal mucosal cell proliferation were performed: immunohistochemical determination of proliferating cell nuclear antigen and whole crypt mitotic count. We found no statistically significant changes in either of these proliferation measures as a result of high or low dairy intake. There was no correlation between the labeling index for proliferating cell nuclear antigen and whole crypt mitotic count; however, measures of the location and intensity of cell proliferation within the rectal crypt were highly correlated between the two assays. Thus, our study indicates that greater consumption of dairy products over a 12-week period does not change rectal mucosal cell proliferation. PMID- 9752984 TI - Aromatic DNA adducts in human white blood cells and skin after dermal application of coal tar. AB - A group of eczema patients topically treated with coal tar (CT) ointments was used as a model population to examine the applicability of DNA adducts in WBC subpopulations as a measure of dermal exposure to polycyclic aromatic hydrocarbons (PAHs). Aromatic DNA adducts were examined by 32P-postlabeling in exposed skin and WBC subsets, and urinary excretion of PAH metabolites was determined to assess the whole-body burden. The median urinary excretion of 1 hydroxypyrene and 3-hydroxybenzo(a)pyrene was 0.39 (range, 0.12-1.57 micromol/mol creatinine) and 0.01 micromol/mol creatinine (range, <0.01-0.04 micromol/mol creatinine), respectively, before the dermal application of CT ointments. After treatment for 1 week, these levels increased to 139.7 (range, 26.0-510.5 micromol/mol creatinine) and 1.18 micromol/mol creatinine (range, <0.01-2.14 micromol/mol creatinine), respectively, indicating that considerable amounts of PAHs were absorbed. Median aromatic DNA adduct levels were significantly increased in skin from 2.9 adducts/10(8) nucleotides (nt; range, 0.7-10.0 adducts/10(8) nt) before treatment to 63.3 adducts/10(8) nt (range, 10.9-276.2 adducts/10(8) nt) after treatment with CT, in monocytes from 0.28 (range, 0.25 0.81 adducts/10(8) nt) to 0.86 adducts/10(8) nt (range, 0.56-1.90 adducts/10(8) nt), in lymphocytes from 0.33 (range, 0.25-0.89 adducts/10(8) nt) to 0.89 adducts/10(8) nt (range, 0.25-3.01 adducts/10(8) nt), and in granulocytes from 0.28 (range, 0.25-0.67 adducts/10(8) nt) to 0.54 adducts/10(8) nt (range, 0.25 1.58 adducts/10(8) nt). A week after stopping the CT treatment, the DNA adduct levels in monocytes and granulocytes were reduced to 0.38 (range, 0.25-0.71 adducts/10(8) nt) and 0.38 adducts/10(8) nt (range, 0.25-1.01 adducts/10(8) nt), respectively, whereas the adduct levels in lymphocytes remained enhanced [1.59 adducts/10(8) nt (range, 0.25-2.40 adducts/10(8) nt)]. Although the adduct profiles in skin and WBC subsets were not identical, and the adduct levels in WBCs were significantly lower as compared with those in skin, the total DNA adduct levels in skin correlated significantly with the adduct levels in monocytes and lymphocytes, but not with those in granulocytes. Excretion of urinary metabolites during the first week of treatment was correlated with the percentage of the skin surface treated with CT ointment and decreased to background levels within a week after the cessation of treatment. 3 Hydroxybenzo(a)pyrene excretion, but not that of 1-hydroxypyrene, correlated significantly with the levels of DNA adducts in skin that comigrated with benzo(a)pyrene-diol-epoxide-DNA. This study indicates that the DNA adduct levels in mononuclear WBCs can possibly be used as a surrogate for skin DNA after dermal exposure to PAHs. PMID- 9752985 TI - Urinary total isothiocyanate (ITC) in a population-based sample of middle-aged and older Chinese in Singapore: relationship with dietary total ITC and glutathione S-transferase M1/T1/P1 genotypes. AB - Isothiocyanates (ITCs), degradation products of glucosinolates (which occur naturally in a variety of cruciferous vegetables), have been shown to exhibit chemopreventive activity. These compounds are metabolized in vivo to form the corresponding dithiocarbamates, which are the major urinary metabolites of ITCs, by a pathway involving the glutathione S-transferase (GST) class of enzymes. Using a newly developed assay that measures total ITC (primarily ITC conjugates) in urine, we examined the relationships between cruciferous vegetable intake (obtained from a food frequency/portion size questionnaire administered in person); dietary total ITC level; GSTM1, GSTT1, and GSTP1 genotypes; and levels of total ITC in spot urine samples collected from 246 Singapore Chinese (111 men and 135 women), ages 45-74 years, who are participants of the Singapore Cohort Study on diet and cancer. Consumption level of cruciferous vegetables was high in study subjects (mean consumption = 345 times per year, mean daily intake = 40.6 g), which was >3 times the comparable level of intake in the United States. Mean daily intake of total ITC among study subjects was 9.1 micromol, and there was a 2.5-fold difference between the 25th and 75th percentile values. Seventy-three % of study subjects tested positive for ITC in urine, and there was a 4-fold difference between the 25th and 75th percentile values among the positive subjects. There was a highly significant positive association between dietary intake and urinary excretion levels of total ITC (two-sided P = 0.0003) that was stronger than the association between overall cruciferous vegetable intake and urinary ITC level, which also was statistically significant (P = 0.0004). There was no difference in urinary ITC levels between GSTM1-null and GSTM1-positive study subjects (P = 0.61) or between subjects with differing GSTP1 genotypes (P = 0.77), but urinary excretion of ITC was significantly higher among GSTT1-positive subjects, relative to GSTT1-null subjects (P = 0.006). The strength of the association between GSTT1 genotype and urinary total ITC level was highly dependent on the level of cruciferous vegetable consumption (or dietary ITC level) in study subjects. Among subjects in the lowest tertile of cruciferous vegetable intake, there was little evidence of an association between GSTT1 genotype and urinary total ITC level (P = 0.67). In contrast, there was a strong and statistically significant association between GSTT1 genotype and urinary total ITC among subjects in the highest tertile of cruciferous vegetable intake (P = 0.02), whereas those in the middle tertile of cruciferous vegetable consumption exhibited an association of intermediate strength (P = 0.04). These results suggest the presence of GSTT1 inducers in cruciferous vegetables. PMID- 9752986 TI - Menstrual factors in relation to breast cancer risk. AB - We evaluated menstrual factors in relation to breast cancer risk in a large, population-based, case-control study. Case women were ascertained through state wide registries covering Wisconsin, Western Massachusetts, Maine, and New Hampshire; control women were randomly selected from driver's license and Medicare lists in each state. Information regarding menstrual characteristics was obtained through a telephone interview. The study population comprised 6888 breast cancer cases and 9529 control women. Because exogenous hormones influence menstrual cycle patterns, we repeated our analyses in a subgroup of women who had never used oral contraceptives or hormone replacement therapy. Our results indicate decreased breast cancer risk with menarcheal age of 15 years or more, relative to menarche at age 13; the relation was stronger among premenopausal [odds ratio (OR), 0.72; 95% confidence interval (CI), 0.57-0.91] as opposed to postmenopausal women (OR, 0.90; 95% CI, 0.80-1.03). Risk was slightly reduced among premenopausal women whose menstrual cycles did not become regular until at least 5 years after onset of menses, relative to those whose cycles became regular within 1 year (OR, 0.80; 95% CI, 0.63-1.02). There was no clear relation between breast cancer risk and irregular menstrual cycles, episodes of amenorrhea, or menstrual cycle length. Early menopause, whether natural or surgical, was associated with decreased breast cancer risk; surgical menopause before age 40 conferred the strongest protective effect (OR, 0.57; 95% CI, 0.47 0.71). We found no evidence of increased risk with late natural menopause (OR, 0.92; 95% CI, 0.80-1.06). Results in the subgroup of women who never used exogenous hormones were similar to those for the entire group. PMID- 9752987 TI - Serological detection of heat shock protein hsp27 in normal and breast cancer patients. AB - Heat shock protein Mr 27,000 (hsp27) is found in many human breast cancer cells and tissues; its expression is associated with the presence of estrogen receptors, lower cell proliferation, and resistance to certain chemotherapies. The purpose of this study was to assess whether hsp27 may be present in sera from women with primary breast cancer and to know whether autoantibodies to hsp27 may be found in these patients. The study was performed by Western blot analyzing sera from 42 normal premenopausal women, 20 normal postmenopausal women, and 36 breast cancer patients. hsp27 was clearly detected in sera by immunoblotting but only after immunoprecipitation. The mean hsp27 levels in cancer patients were higher than in the control patients; however, 66% of the breast cancer patients showed hsp27 within the normal range, indicating low sensitivity. Moreover, cancer patients with metastatic disease did not show significantly higher hsp27 levels than cancer patients without metastases. Serum hsp27 levels did not correlate with the hsp27 levels in tumor tissues detected by immunohistochemistry. Elevated CA 15-3 levels were not associated with high hsp27 values. Autoantibodies against hsp27 were not detected by immunoblotting in normal sera and in sera from breast cancer patients. As a consequence, serological determination of this biomarker is unlikely to be of utility in the detection and follow-up of breast cancer patients. PMID- 9752988 TI - Farm and animal exposures and pediatric brain tumors: results from the United States West Coast Childhood Brain Tumor Study. AB - Nineteen counties from San Francisco and Los Angeles, California and Seattle, Washington were the United States sites for a large population-based case-control study of childhood brain tumors (CBTs), sponsored by the National Cancer Institute. CBT patients who were < 20 years of age and were diagnosed between 1984 and 1991 were reported to each region's cancer registry. The 801 control subjects were obtained by random digit dial and were frequency-matched to the 540 CBT patients in San Francisco and Seattle (one patient to two controls) and in Los Angeles (one patient to one control). Data collected by in-person interview with subjects' mothers were analyzed to investigate an association between risk for CBTs and life on a farm, exposure to farm animals (dairy cattle, beef cattle, pigs, sheep/goats, poultry, and horses), and some cat and non-farm horse exposures. Elevated risks for CBTs were observed in association with mothers' exposure to pigs [odds ratio (OR) = 3.8, 95% confidence interval (CI) = 1.2-12] and horses (OR = 2.2, 95% CI = 1.0-4.8) on a farm during the index pregnancy. Children diagnosed with primitive neuroectodermal tumors showed elevated risks for CBTs with personal and maternal prenatal exposure to pigs (child, OR = 4.0, 95% CI = 1.2-13; mother, OR = 11.9, 95% CI = 2.8-51) and poultry (child, OR = 3.0, 95% CI = 1.1-8.0; mother, OR = 4.0, 95% CI = 1.2-14). No other animal exposures of children or mothers were found to be consistently related to CBTs. Children diagnosed with primitive neuroectodermal tumors who were on a farm for > 1 year and were first on a farm when they were < 6 months of age also had increased risk for CBTs (OR = 3.9, 95% CI = 1.2-13). A somewhat increased risk for CBTs was found for children of mothers who ever had worked on livestock farms compared with mothers who never had worked on a farm (OR = 7.4, 95% CI = 0.86-64, based on five case mothers and one control mother who worked on livestock farms during the 5 years preceding the birth of the index child). The associations are consistent with those of two previous studies in Norway (P. Kristensen et al., Int. J. Cancer, 65: 39-50, 1996) and the United States and Canada (G. R. Bunin et al., Cancer Epidemiol. Biomark. Prev., 3: 197-204, 1994) that investigated the role of farm-related exposures in the etiology of CBTs. PMID- 9752989 TI - Quality control program for storage of biologically banked blood specimens in the Malmo Diet and Cancer Study. AB - A biological bank has been developed to extend the biochemical and molecular research base for a prospective study on diet and cancer in the city of Malmo, Sweden. The study entered individuals 45-69 years of age, of which 30,382 individuals (45%) participated. Each individual entering the bank has stored samples of viable mononuclear leukocytes (MNLs; -140 degrees C) and granulocytes (GRANs; -80 degrees C) or buffy coats (-140 degrees C), erythrocytes (-80 degrees C), and plasma/serum (-80 degrees C). The bioassays developed to monitor the quality of storage conditions were: (a) viability and growth response to phytohemagglutinin for MNLs; (b) DNA strand breakage for GRANs; (c) NAD content for erythrocytes; and (d) thiol status for plasma/serum. The yield, purity, and storage conditions were all quality controlled, and the samples were determined to be of high standard after 137-190 weeks of storage. No differences in yield and purity were found in samples banked by different laboratory technicians. Growth responses of MNLs were severely reduced (90%) after 40 weeks of storage, which justified switching from the storage of purified MNLs and GRANs to the more cost-effective banking of buffy coats. We conclude that the quality of the banked material, based on the biochemical analysis done, indicate that the storage conditions are optimal at least up to 3.5 years, except for the growth response of MNLs. PMID- 9752990 TI - Feasibility and quality of biological banking of human normal and tumor tissue specimens as sources of DNA for the Malmo Diet and Cancer Study. AB - Human tumor and normal tissue specimens, which were collected from autopsy material 1-6 days postmortem, were compared with similar tissue specimens collected within 2 h after surgical resection and transport to the pathology department. The end point criteria used to evaluate the quality of the specimens for biological banking purposes were the extractability and yield of high molecular weight DNA and UV absorption ratios at 260:280 after collection and immediate storage of the specimens at -80 degrees C. The data demonstrated that autopsy material was a quality source of DNA, although of not such high quality as surgical biopsy specimens <2 h after resection. The advantages of using autopsy material to supplement surgical specimen collection sent to pathology, as opposed to using specimen collection at surgery wards or formalin-fixed material, as sources of DNA are: (a) large amounts of tumor and normal tissues from a variety of organ sites can be obtained without regard to the patient's health status; (b) a higher percentage of retrieval of incident cases of cancer in prospective designed trials is more likely to be achieved; and (c) the extractable DNA is of sufficiently high enough quality to permit direct analyses by molecular hybridization and sequence methodologies. PMID- 9752991 TI - Design and baseline characteristics of study participants in the Wheat Bran Fiber trial. AB - The Wheat Bran Fiber (WBF) trial is a Phase III clinical trial designed to assess the effect of a WBF intervention for 3 years on the recurrence of adenomatous polyps. Men and women, 40-80 years of age, who had removal of one or more colorectal adenoma(s) 3 mm or larger within 3 months prior to study entry were recruited from three sites in the Phoenix metropolitan area. After meeting eligibility criteria, 1509 individuals entered a 6-week run-in period, consisting of a low WBF (2 g/day) intervention. Participants (n = 1429) successfully completed this phase and were randomized to a high (13.5 g/day) or low (2 g/day) WBF intervention. Various data and specimens were collected at baseline and throughout the intervention phase, which included dietary intake, physical activity, other risk factor information, blood specimens, rectal biopsies, and polyp tissues. The study design called for a colonoscopy at approximately 1 year after the qualifying colonoscopy; thus, the period between the first year and the final colonoscopy will be used to assess the effect of the intervention, which is expected to be completed in the latter part of 1998. PMID- 9752992 TI - 4-Hydroxy-1-(3-pyridyl)-1-butanone-hemoglobin adducts as biomarkers of exposure to tobacco smoke: validation of a method to be used in multicenter studies. AB - Hemoglobin (Hb) adducts of 4-hydroxy-1-(3-pyridyl)-1-butanone (HPB), a metabolite of two tobacco-specific nitrosamines [4-(methylnitrosamino)-1-(3-pyridyl)-1 butanone and N'-nitrosonornicotine], were measured as biomarkers of exposure to tobacco smoke as part of a study on genetic alterations and susceptibility to lung cancer among nonsmokers. HPB-Hb adducts were measured after collection of RBCs by Ficoll gradient in six collaborating centers, release of HPB by alkaline hydrolysis from Hb, clean-up by solid-phase extraction, and analysis of an electron-capturing derivative by gas chromatography-electron capture mass spectrometry. Prior to analysis of samples from study subjects, the reproducibility of this approach was validated in blood from donors. The coefficient of variation of reproducibility of paired aliquots from five samples ranged from 7 to 25%; the within-sample reproducibilities of four and eight aliquots were 4 and 16%, respectively. The study subjects consisted of 18 smokers and 52 never-smokers. HPB-Hb adduct levels were significantly higher (P = 0.02) in smokers (26 +/- 13 fmol HPB/g Hb) than in never-smokers (20 +/- 8 fmol HPB/g Hb). There was no difference between sexes. These results suggest that the level of HPB-Hb adducts, measured using a method modified to facilitate use in multicenter studies, can be a useful biomarker of exposure to tobacco smoke. PMID- 9752993 TI - Cytochrome P450 1A1 MspI polymorphism and urinary 1-hydroxypyrene concentrations in coke-oven workers. AB - Coke-oven workers are regularly exposed to a high concentration of the benzene soluble fraction (BSF) of total particulates, which are comprised mainly of polycyclic aromatic hydrocarbons (PAHs). A metabolite of pyrene, 1-hydroxpyrene (1-OHP), is readily measured in the urine of exposed individuals. Epidemiological studies have shown that P4501A1 (CYP1A1) genotypes are associated with PAH related lung cancer risk. The purpose of this study was to investigate whether CYP1A1 MspI genotypes modulate the relationship of individual occupational exposure to air BSF to urinary 1-OHP concentrations among coke-oven workers. We monitored individual breathing zone air BSF over 3 consecutive days in 80 coke oven workers in Taiwan from August 1995 to February 1996. Exposure was also dichotomized by work area (topside oven workers and sideoven workers). Preshift urine on the morning of day 1 and postshift urine on the afternoon of day 3 were measured by fluorescent spectrophotometry, and blood samples were analyzed to determine the relative distributions of CYP1A1 MspI polymorphisms. The frequency of the MspI homozygous variant genotypes of CYP1A1 was 15%. Multiple linear regression showed significant effects of individual occupational exposure to air BSF and preshift 1-OHP on postshift urinary 1-OHP concentrations (P = 0.002 and P < 0.001, respectively). After adjusting for preshift 1-OHP concentrations and air BSF, subjects with the homozygous variant genotype have a 2-fold higher postshift 1-OHP levels than the combined wild-type and heterozygous (P = 0.04). In addition, a positive trend was found in postshift 1-OHP and across-shift change of 1-OHP (postshift 1-OHP - preshift 1-OHP) in decreasing order, as follows: topside oven workers with the homozygous variant trait, topside oven workers with the heterozygous variant trait, sideoven workers with the homozygous variant trait, and sideoven workers with the heterozygous trait (P < 0.001). We conclude that CYP1A1 MspI variant genotype can modify the metabolism of PAHs in coke-oven workers. PMID- 9752994 TI - Childbearing and the risk of Hodgkin's disease. AB - The causes of Hodgkin's disease remain incompletely known, but a higher incidence in men than in women has prompted an interest in the role of female sex hormones and reproductive history. Available epidemiological data are, however, contradictory. We analyzed possible associations between parity, age at first birth, and the risk of developing Hodgkin's disease by a linkage between the Swedish Cancer Register and a nationwide Fertility Register. Among women born between 1925 and 1972, 917 cases with Hodgkin's disease and concomitant fertility information were identified. For each case patient, five age-matched controls were randomly selected among women in the Fertility Register. Conditional logistic regression was used to estimate odds ratios of Hodgkin's disease associated with a birth. We found a slightly and nonsignificantly reduced risk of Hodgkin's disease in ever-parous compared with nulliparous women. Among parous women, the number of children was unrelated to risk, whereas there was some evidence of an increased risk with late age at first birth in women under age 45 at diagnosis. No clear temporal relations between childbearing and subsequent risk were discernible in any parity or age group. Although uncontrolled confounding might have affected our results, they do not indicate that hormonal or immunological changes associated with childbearing play a role in the development of Hodgkin's disease. PMID- 9752995 TI - Birthplace and yield of nipple aspirate fluid in Chinese women. AB - The different rates of breast cancer found between Chinese women in Asia compared with Chinese-born women in the United States suggest that dietary and environmental factors may be of etiological significance. We evaluated the proportion of 480 premenopausal Chinese women who yielded nipple aspirate fluid (NAF) by birthplace in Asia versus the United States and by reproductive and other risk factors. Birthplace was used as a surrogate for presumed differences in exposures during gestation, childhood, and adolescence that might influence yield of NAF in premenopausal women. In United States-born Chinese women compared with Asia-born Chinese women, the proportion yielding NAF was 44 of 95 (46.3%) versus 120 of 385 (31.2%), respectively. The relative risk of yield of NAF in United States-born women compared with Asia-born women was odds ratio = 2.37 (95% confidence interval, 1.26-4.47). Independent positive associations of NAF yield were also found with history of parity and breast feeding, cerumen phenotype, and a negative association with ever use of oral contraceptives. These findings support the hypothesis that early environmental exposures may have long-lasting physiological effects discernible in the breast glands of adult women. PMID- 9752996 TI - Immunological and chemical studies of Salmonella haarlem somatic antigen epitopes. I. Structural studies of O-antigen. AB - Lipopolysaccharide (LPS) of Salmonella haarlem was hydrolyzed and the products separated. The structure of the O-specific polysaccharide (OPS) was found from sugar and methylation analyses. Rhamnose, mannose, galactose and tyvelose were detected and their linkage modes were established. The structure was confirmed by 1H, homonuclear and heteronuclear correlations and 13C NMR spectra. Anomeric configurations were assigned by chromium trioxide oxidation and proton coupled 13C spectra. Sugar sequence was established from specific carbon shift data and nuclear Overhauser effect spectroscopy. The repeating unit structure of S. haarlem OPS as --> 3)-alpha-D-Galp-(1 --> 6)-[alpha-Tyvp-(1 --> 3)]-beta-D-Manp (1 --> 4)-alpha-L-Rhap was estimated. No structural heterogeneity of the antigen was found. PMID- 9752997 TI - Immunological and chemical studies of Salmonella haarlem somatic antigen epitopes. II. Serological investigations. AB - Lipopolysaccharide of Salmonella haarlem was hydrolyzed and the products separated. Native O-polysaccharide antigen was oxidised with sodium periodate followed by reduction with sodium borohydride. Native and chemically modified antigens were the subject of immunochemical studies. Monoclonal antibodies against S. haarlem and polyclonal rabbit antisera against S. haarlem, S. typhi and S. anatum bacteria were produced. The serological relationship between the lipopolysaccharide of Salmonella bacteria belonging to the two different groups D2 and E1 was demonstrated using haemagglutination reactions, inhibition of haemagglutination and immunoblotting. Cross-reactions were observed in haemagglutination reactions and in immunoblotting between antisera to S. haarlem, S. typhi and S. anatum. Factors 3,9,46 were found in the S. haarlem strain and the sugar composition for the epitopes of each factor was determined. PMID- 9752998 TI - Identification of S, F1C and three PapG fimbrial adhesins in uropathogenic Escherichia coli by polymerase chain reaction. AB - S and F1C fimbrial adhesins often expressed by uropathogenic Escherichia coli are genetically homologous. A multiply primed polymerase chain reaction (PCR) was developed for discriminating the S (sfa) and F1C (foc) fimbrial operons. A total of 270 uropathogenic E coli strains and 80 fecal isolates were examined. PCR specifically detected the sfa and foc alleles in 105 (93%) of 113 sfa/foc+ strains by DNA hybridization. Furthermore, 87% of sfa+ uropathogenic E. coli simultaneously possessed the genes encoding the class III P fimbrial adhesin (prsG(J96)), alpha-hemolysin and cytotoxic necrotizing factor 1. Statistical analysis showed the class II P fimbrial adhesin (papG(IA2)) and F1C fimbria to be associated with high relative virulence in pyelonephritis and cystitis, respectively. The multiply primed PCR developed should be useful for assessing the contribution of the S and F1C fimbriae in the pathogenesis of urinary tract infections. PMID- 9752999 TI - Cloning of a sapB homologue (sapB2) encoding a putative 112-kDa Campylobacter fetus S-layer protein and its use for identification and molecular genotyping. AB - A sap gene encoding a surface layer protein was isolated from a Campylobacter fetus ssp. fetus CIP 53.96T cosmid library. This sap gene, which shows significant homology with the sapB conserved region, was named sapB2. The complete ORF of 3339 nucleotides encodes a 1112-amino acid polypeptide with a calculated molecular mass of 112 kDa. High homology with the sapB gene was found in a region beginning 67 bp before the ORF and proceeding 546 bp into the ORF. Similarly, 98% homology with the sapA2 gene was observed in a 2038-bp region beginning 540 bp after the initiation codon. In the present study, we show that this sapB2 gene has two main interesting features: the 5' end of the region which presents high homology with the sapA2 homologue was found to be present in every C. fetus strain, and the fragment (IG01) comprising the region which presents homology with the sapB conserved region and the 5' end of the sapA2 homologue region, when used as a probe, can reveal genomic polymorphism among C. fetus strains. We exploited these features to develop a PCR assay for the specific detection of C. fetus and to set up a method for typing C. fetus isolates. The PCR assay was found to be species-specific. Oligonucleotide primers derived from the 5' end of sapA2 homologue region were used in a polymerase chain reaction test on genomic DNA extracted from 101 Campylobacter fetus, 18 Campylobacter non fetus and seven non-Campylobacter strains. A 220-bp fragment was amplified only when C. fetus DNA was used as a target. In Southern blot analysis, the IG01 probe was found to hybridize only with DNA extracted from C. fetus strains. Moreover, IG01 hybridized with several fragments of HindIII-digested DNA, giving a specific pattern for each strain. PMID- 9753000 TI - Macrophage-lymphocyte interactions mediate anti-Burkholderia pseudomallei activity. AB - The mechanisms involved in the pathogenesis of melioidosis, caused by the intracellular bacterium Burkholderia pseudomallei, are unclear. C57BL/6 mice are resistant to infection, while BALB/c mice are highly susceptible. Previous studies have demonstrated that peritoneal exudate cell preparations enriched for macrophages are capable of effectively eliminating intracellular pathogens. In this study we present evidence showing that interaction of macrophages with lymphocytes is necessary for efficient anti-B. pseudomallei activity. PMID- 9753001 TI - Conditional adherence of Enterococcus faecalis to extracellular matrix proteins. AB - The adherence of 44 clinical isolates of Enterococcus faecalis, a common cause of endocarditis, and 13 Enterococcus faecium to substrates of six extracellular matrix (ECM) proteins was examined using 35S-labeled bacteria. One E. faecalis strain, isolated from a patient with endocarditis, adhered to collagen types I and IV and another E. faecalis strain adhered to laminin and to collagen types I and IV. However, most isolates showed little adherence ( < 5% of added cells adhered) when grown at 37 degrees C regardless of their source (endocarditis, urine or fecal sample). When grown at 46 degrees C (but not when grown in CO2 or nutrient limited media), most isolates of E. faecalis increased their adherence to immobilized laminin, collagen types I and IV but not to fibronectin, fibrinogen or bovine serum albumin, whereas none of the E. faecium increased adherence when grown at 46 degrees C or 50 degrees C. The adherence of E. faecalis was eliminated by digestion with trypsin, suggesting that a protein is somehow important, directly or indirectly, for adherence to occur. Pre-incubation of bacteria with soluble collagen types I and IV inhibited the adherence to these ECM proteins. These results demonstrate that in E. faecalis, adherence to ECM proteins is produced during routine in vitro growth conditions by occasional isolates and can be produced during certain stressful growth conditions by others. Whether this adherence relates to the propensity of E. faecalis to cause endocarditis remains to be determined. PMID- 9753002 TI - Carbapenem-induced endotoxin release in gram-negative bacterial sepsis rat models. AB - The carbapenem-induced endotoxin release was evaluated using experimental models of gram-negative bacterial sepsis in Wistar rats. Infections with Escherichia coli, Serratia marcescens, Klebsiella pneumoniae, Pseudomonas aeruginosa, Proteus vulgaris and Proteus mirabilis resulted in an increase of the plasma endotoxin concentration after treatment with ceftazidime and carbapenems including imipenem, panipenem, meropenem and biapenem. Except for P. aeruginosa, the plasma endotoxin concentrations after carbapenem treatment were significantly lower than those after ceftazidime treatment. It is noteworthy that treatment of P. aeruginosa sepsis with meropenem or biapenem induced significantly more endotoxin release than other carbapenems and the endotoxin concentrations induced by these carbapenems reached those of ceftazidime treatment. The plasma endotoxin concentrations appeared to correlate with the reduction of platelet counts and the elevation of both glutamic oxaloacetic transaminase and glutamic pyruvic transaminase values. PMID- 9753003 TI - Role of CD86 (B7-2) in triggering of antigen-specific IgE antibody response by lipopolysaccharide. AB - The role of CD86 in triggering of ascaris extract-specific IgE antibody response by lipopolysaccharide was studied. The simultaneous administration of anti-CD86 antibody with ascaris extract and lipopolysaccharide prevented the production of IgE antibody response to ascaris extract. CD86+ cells were detected in peritoneal cavities and spleens of mice injected intraperitoneally with lipopolysaccharide. CD86+ cells appeared in peritoneal cavities and spleens eight hours after lipopolysaccharide injection, and they were detectable for a week. CD86+ cells in peritoneal cavities and spleens were mainly surface Ig-positive B-cells and some Ig-negative cells. It was suggested that lipopolysaccharide induced the expression of CD86 mainly on B-cells, and that CD86+ cells induced by lipopolysaccharide injection might play an important role as antigen-presenting cells on triggering of ascaris extract-specific IgE antibody response by lipopolysaccharide. PMID- 9753004 TI - In vitro inhibition of adhesion of enterotoxigenic Escherichia coli K88 to piglet intestinal mucus by egg-yolk antibodies. AB - The objective of the study was to determine if the adhesion of E. coli K88 to piglet intestinal mucus could be inhibited in vitro by spray-dried egg-yolk anti K88 antibodies. Binding of E. coli was monitored using a radioactive assay. Four 14+/-2-day-old healthy piglets were used for the preparation of mucus from the small intestine. Competition and displacement phenomena were investigated by incubating (a) egg-yolk antibodies and E. coli together prior to adding to the mucus and (b) E. coli and mucus, followed by egg-yolk antibodies. The results demonstrated that egg-yolk antibodies inhibited the adhesion of 3H-labeled local strain of hemolytic E. coli K88+ (E. coli K88+ MB) to piglet small intestinal mucus by 84.6-97.0% when the egg-yolk antibodies were diluted 10, 20, 40 or 100 times. The adhesion inhibiting effects of egg-yolk antibodies declined dramatically when the antibody dilution was more than 250-fold. A similar adhesion inhibiting effect was observed when egg-yolk antibodies were incubated with E. coli K88+ MB for 15, 30 and 60 min prior to the adhesion test. Egg-yolk antibodies when diluted 50- and 100-fold had a very strong inhibiting ability against E. coli K88+ MB at a concentration of 10(9) colony forming units (cfu) ml(-1) (adhesion was < 6%). However, dilution of 100 times for egg yolk antibodies was insufficient to inhibit the adhesion of E. coli K88+ MB to intestinal mucus when the concentration of E. coli K88+ MB was 10(10) cfu ml(-1). The displacement test indicated that there was no significant reduction in the adhesion of E. coli K88+ MB to the small intestinal mucus when egg-yolk antibodies were added after adhesion of the organism to the mucus. These studies demonstrate that anti-K88+ MB fimbriae antibodies from chicken egg-yolk when added to E. coli K88+ MB prevented their binding to receptors in the mucus isolated from the intestine of piglets. PMID- 9753005 TI - In vivo testing of an Enterococcus faecalis efaA mutant and use of efaA homologs for species identification. AB - Disruption of the previously described efaA (from Enterococcus faecalis antigen A) gene was generated in E. faecalis strain OG1RF and loss of an 37-kDa immunoreactive band from the mutant was demonstrated in Western blots. In a mouse peritonitis model, mice infected with the efaAfs (fs=from Enterococcus faecalis) mutant showed more prolonged survival than mice infected with the parent strain OG1RF. These results suggest that efaAfs encodes a function important for infection of mice by enterococci. An efaA-like gene was also identified in E. faecium DNA libraries and its deduced amino acid sequence showed 73% similarity to EfaA of E. faecalis and 42-63% similarities to a group of streptococcal virulence and adhesion associated proteins that are components of ATP-binding cassette transport systems. Intragenic probes representing efaAfs, recAfs, efaAfm (fm=from E. faecium) and gyrAfm were tested for their ability to identify E. faecalis and E. faecium using colony lysates of 133 enterococci and one Streptococcus sp. Probes of E. faecium and E. faecalis origin hybridized to all isolates of E. faecium and E. faecalis, respectively, regardless of their clinical source but not to any of 29 other enterococci. These results suggest that the use of gene probes may prove helpful in identification of isolates of E. faecium and E. faecalis. PMID- 9753006 TI - Vitamins and vitriol: W.L. Braddon's epidemiology of Beriberi. PMID- 9753007 TI - Systolic blood pressure trends in US adults between 1960 and 1980: influence of antihypertensive drug therapy. AB - Recent blood pressure trends reflect progress in hypertension control, but prevalent drug therapy precludes direct estimation of the component due to primary prevention. In data gathered on persons aged 35-74 years in three successive US health examination surveys (1960-1980), systolic blood pressure levels assuming no drug therapy were imputed by reassigning blood pressure to the upper end of the distribution for respondents reporting use of antihypertensive medication. Blood pressure was partitioned into four ordinal categories based on weighted percentiles of the 1960-1962 distributions for 35- to 44-year-old males and females who reported no use of antihypertensive medication. Cumulative logit models (alpha = 0.01) were used to estimate age- and sex-specific trends for blacks and whites within two strata (<25 or > or =25) of body mass index (BMI) (weight (kg)/height (m)2). Before imputation, systolic blood pressure decreased between 1960 and 1980; after imputation, significant decreases remained only in 35- to 44-year-olds. Strong associations of black race and BMI > or =25 with higher blood pressures were present in models with and without drug therapy. Thus, according to the models, there has been little progress in decreasing racial or BMI-related blood pressure differentials. Above the age of 44 years, blood pressure trends were largely attributable to medication use. In contrast, data for 35- to 44-year-olds suggest progress in primary prevention. PMID- 9753008 TI - Weight history, glucose intolerance, and insulin levels in middle-aged Swedish men. AB - The association between weight history and glucose intolerance was examined in a cross-sectional study consisting of 3,128 Swedish men aged 35-56 years, 52 percent of whom had a family background of diabetes mellitus. Oral glucose tolerance testing detected 55 cases of type 2 (non-insulin-dependent) diabetes and 172 cases of impaired glucose tolerance. Among men with no family history of diabetes, the estimated odds ratios for impaired glucose tolerance associated with short (<5 years) and long (> or =10 years) durations of obesity (body mass index (weight (kg)/height2 (m2) > or =25.0) were 1.3 (95% confidence interval (CI) 0.2-7.7) and 11.8 (95% CI 3.3-41.9), respectively. Among men with a family history of diabetes, the odds ratios were 2.0 (95% CI 0.8-4.7) and 4.0 (95% CI 1.8-9.1), respectively. Corresponding estimates of the odds of type 2 diabetes, adjusted for family history of diabetes, were 1.9 (95% CI 0.5-7.1) and 7.3 (95% CI 2.2-23.7), respectively. The odds of high (> or =30.0 mU/liter) fasting insulin levels in subjects with impaired glucose tolerance were 6.9 (95% CI 0.6 74.2) and 21.0 (95% CI 2.1-206.4) for short and long durations of obesity, respectively. Corresponding estimated odds of low 2-hour insulin response (< or =71.9 mU/liter) were 0.7 (95% CI 0.2-2.9) and 3.3 (95% CI 1.2-8.9). Homeostasis model assessment of insulin resistance yielded an odds ratio of 6.7 (95% CI 0.6 73.4) for a short duration of obesity and 20.0 (95% CI 2.0-200.6) for a long duration. Examination of beta-cell function with homeostasis model assessment resulted in odds ratios of 0.2 (95% CI 0.0-1.6) and 2.0 (95% CI 0.7-5.4) for short and long durations of obesity, respectively. These data indicate that obesity decreases glucose tolerance by way of progressively increased insulin resistance and, in the case of prolonged duration, by decreased insulin secretion as well. PMID- 9753009 TI - Voluntary and involuntary weight loss: associations with long term mortality in 9,228 middle-aged and elderly men. AB - Recent studies have suggested that weight loss in middle-aged persons antecedes increased mortality. Therefore, the authors sought to examine the association between changes in body weight and subsequent mortality, according to self reported dieting status. The authors followed 9,228 men aged 40-65 years in 1963, for whom weight changes between 1963 and 1968 were recorded and extensive clinical, anthropometric, biochemical, and dietary assessments were made. Of these men, 2,471 reported being on a diet when first examined in 1963, and 636 were dieting primarily to lose weight. Mortality follow-up covered an 18-year period (1968-1986). Men who lost 5 kg or more between 1963 and 1968 ("extreme weight losers") exhibited the following age-pooled risks of mortality relative to the stable weight group: for total mortality, 1.36 (95% confidence interval (CI) 1.20-1.55); for all cardiovascular disease mortality, 1.40 (95% CI 1.16-1.69); for all non-cardiovascular disease mortality, 1.33 (95% CI 1.11-1.59); for coronary heart disease mortality, 1.55 (95% CI 1.25-1.93); and for cancer mortality, 0.90 (95% CI 0.65-1.24). After adjustment for differences in coronary heart disease risk factor levels and morbidity between these groups at the end of the weight change period (1968), the excess risks associated with extreme weight loss declined by approximately one third. They declined further if adjustment was made for 1963 (pre-weight-change period) morbidity and risk factor levels. Being on a slimming diet, as reported in 1963, was associated with an approximate doubling of excess mortality in men with extreme weight loss. Weight loss in 1963 1968 coincided with an increased incidence of coronary heart disease and diabetes mellitus and a declining level of serum total cholesterol. This and other studies indicate that both voluntary and involuntary weight loss might be associated with a small increase in the risk of all-cause mortality. PMID- 9753010 TI - Electric blanket use and breast cancer risk among younger women. AB - To investigate whether use of electric blankets, one of the largest sources of electromagnetic field exposure in the home, is associated with the risk of female breast cancer, the authors analyzed data from a population-based US case-control study. The 2,199 case patients were under age 55 years and had been newly diagnosed with breast cancer between 1990 and 1992. The 2,009 controls were frequency-matched to cases by 5-year age group and geographic area. There was little or no risk associated with ever having used electric blankets, mattress pads, or heated water beds among women under age 45 years (adjusted odds ratio = 1.01, 95% confidence interval 0.86-1.18) or among women aged > or =45 years (adjusted odds ratio = 1.12, 95% confidence interval 0.87-1.43). There was no substantial variation in risk with duration of use; with whether the appliance was used only to warm the bed or used throughout the night; with menopausal status; or with the cases' hormone receptor status or stage of disease. Potential breast cancer risk factors that were associated with electric blanket use did not substantially confound the associations under investigation. These data do not support the hypothesis that electric blanket use increases breast cancer risk among women under age 55 years. PMID- 9753011 TI - First analysis of mortality and occupational radiation exposure based on the National Dose Registry of Canada. AB - A cohort mortality study of occupational radiation exposure was conducted using the records of the National Dose Registry of Canada. The cohort consisted of 206,620 individuals monitored for radiation exposure between 1951 and 1983 with mortality follow-up through December 31, 1987. A total of 5,426 deaths were identified by computerized record linkage with the Canadian Mortality Data Base. The standardized mortality ratio for all causes of death was 0.61 for both sexes combined. However, trends of increasing mortality with cumulative exposure to whole body radiation were noted for all causes of death in both males and females. In males, cancer mortality appeared to increase with cumulative exposure to radiation, without any clear relation to specific cancers. Unexplained trends of increasing mortality due to cardiovascular diseases (males and females) and accidents (males only) were also noted. The excess relative risk for both sexes, estimated to be 3.0% per 10 mSv (90% confidence interval 1.1-4.8) for all cancers combined, is within the range of risk estimates previously reported in the literature. PMID- 9753012 TI - Smoking after age 65 years and mortality in Barcelona, Spain. AB - The objective of this study was to assess the risk of dying associated with smoking after the age of 65 years and the benefits of quitting smoking, taking into account baseline health status. The study was carried out in Barcelona, Spain, a southern European city with an increase in smoking prevalence and lifestyle different from those of other areas where hazards of smoking have been studied. A follow-up study begun in 1986 was carried out in 477 males (94.3% of the original cohort) who were randomly selected by census from members of the Barcelona general population aged > or =65 years. Vital status as of October 1994 and, where applicable, cause of death (cardiovascular disease, cancer, or respiratory disease) were assessed. The relative risk of dying was 2.11 (95% confidence interval (CI) 1.37-3.26) times higher in current smokers and 1.53 (95% CI 1.03-2.27) times higher in former smokers than in never smokers. Quitting smoking after the age of 65 years reduced the relative risk of dying to 0.77 (95% CI 0.51-1.16) in comparison with continuing to smoke, although persons who stopped smoking had poorer self-perceived health and were more frequently reported to suffer from cardiovascular disease (p < 0.05). This study confirms that the effects of smoking extend to later life in this elderly general population, with a magnitude as great as that seen in previous studies with different populations. In addition, it indicates that stopping smoking after age 65 reduces the risk of dying. PMID- 9753013 TI - Socioeconomic status differences in hormone therapy. AB - Socioeconomic status (SES) is a significant sociodemographic correlate of noncontraceptive hormone therapy, yet multiple dimensions of SES have not been examined systematically in previous studies of hormone therapy. This study examined the lifetime incidence of noncontraceptive hormone therapy, how usage varied by type of reproductive organ surgery, and the bivariate and net associations between a large array of SES indicators and the likelihood of ever using hormones by age 53-54 years in a population sample (n = 3,612) of non Hispanic white female participants in the Wisconsin Longitudinal Study (1957 1993). Approximately half of the women had ever used noncontraceptive hormones; 38.5% were current users. In multivariate logistic regression analyses, the most robust SES predictor of hormone therapy was a woman's husband's occupational status (higher status associated with higher rates of use), after adjustment for all other measured sociologic and biomedical factors (e.g., other SES measures, other health behaviors, menopausal symptoms, age at menopause, health insurance). The association of hormone therapy with education differed between women who underwent hysterectomy and/or oophorectomy (higher odds for less educated women) and those with intact reproductive organs (lower odds for less educated women). Additionally, a woman's own earnings and household net worth were positive net correlates of noncontraceptive hormone therapy. PMID- 9753014 TI - Dietary vitamin C intake and lung function in rural China. AB - The relation between dietary vitamin C intake and pulmonary function was investigated in a cross-sectional study carried out in 69 counties in rural China in 1989. Within each of the 69 counties, 120 subjects aged 35-64 years were identified using a three-stage random clustering procedure. Each subject underwent pulmonary function testing, completed a detailed questionnaire, and provided a blood sample. Dietary vitamin C intakes were estimated among half of the subjects using a 3-day weighed record of household food intake. Plasma vitamin C was measured in sex-specific blood pools created from individual samples in each geographic area. Among the 3,085 subjects for whom there were complete data, dietary intake of vitamin C (151 mg/day (standard deviation, 111)) was significantly related to forced expiratory volume in the first second (FEV1) and forced vital capacity after adjustment for sex, age, height, weight, total caloric intake, tobacco smoking, and education. An increase of 100 mg/day in vitamin C intake was associated with an increase of 21.6 ml (95% confidence interval -0.4 to 43.5) in FEV1 and an increase of 24.9 ml (95% confidence interval 0.2 to 49.6) in forced vital capacity. No significant interaction with smoking status was observed. A significant positive association was also observed at the geographic level, between county-pooled plasma vitamin C and mean FEV1. These data support the hypothesis that dietary vitamin C may protect against the loss of pulmonary function. PMID- 9753015 TI - Use of transition probabilities to estimate the effect of smoking on the duration of episodes of respiratory symptoms in diary data: the Swiss Study on Air Pollution and Lung Diseases in Adults (SAPALDIA). AB - Incompletely documented symptom episodes pose methodological problems in the analysis of diary data. The aim of this study was to develop a method of estimating the average durations of symptomatic and nonsymptomatic episodes, respectively, coping with the problem of bias due to undocumented days and censored episodes that is found in most diary studies. The authors derived their outcome variables from a Markov model using transition probabilities. To evaluate this method, the authors assessed the impact of active smoking on the duration of episodes of bronchitis symptoms and the corresponding nonsymptomatic periods, respectively, using diary data (1992-1993) obtained from 801 participants in the Swiss Study on Air Pollution and Lung Diseases in Adults. Covariate-adjusted distribution curves for the mean durations of individual episodes were estimated by Cox regression. Median values for light smokers (<10 cigarettes/day) were 60.0 symptom-free days (95% confidence interval (CI) 42.0-78.5) and 4.0 symptomatic days (95% CI 3.0-6.0), respectively, compared with medians of only 21.0 days (95% CI 16.2-29.8) for periods without bronchitis symptoms and 6.0 days (95% CI 4.9 9.0) for episodes of bronchitis symptoms in heavy smokers (> or =30 cigarettes/day). The authors suggest that the Markov method is a feasible approach to the assessment of long term effects of smoking and environmental risk factors on the average duration of symptomatic and nonsymptomatic respiratory episodes. PMID- 9753016 TI - Estimating sensitivity and sojourn time in screening for colorectal cancer: a comparison of statistical approaches. AB - The effectiveness of cancer screening depends crucially on two elements: the sojourn time (that is, the duration of the preclinical screen-detectable period) and the sensitivity of the screening test. Previous literature on methods of estimating mean sojourn time and sensitivity has largely concentrated on breast cancer screening. Screening for colorectal cancer has been shown to be effective in randomized trials, but there is little literature on the estimation of sojourn time and sensitivity. It would be interesting to demonstrate whether methods commonly used in breast cancer screening could be used in colorectal cancer screening. In this paper, the authors consider various analytic strategies for fitting exponential models to data from a screening program for colorectal cancer conducted in Calvados, France, between 1991 and 1994. The models yielded estimates of mean sojourn time of approximately 2 years for 45- to 54-year-olds, 3 years for 55- to 64-year-olds, and 6 years for 65- to 74-year-olds. Estimates of sensitivity were approximately 75%, 50%, and 40% for persons aged 45-54, 55 64, and 65-74 years, respectively. There is room for improvement in all models in terms of goodness of fit, particularly for the first year after screening, but results from randomized trials indicate that the sensitivity estimates are roughly correct. PMID- 9753018 TI - What managed care organizations want from internal medicine training programs. PMID- 9753017 TI - Re: "Reliability of reported sunburn history in a case-control study of cutaneous malignant melanoma". PMID- 9753019 TI - Efficacy and safety of an early discharge protocol in low-risk patients with upper gastrointestinal bleeding. AB - PURPOSE: The outcome of patients with upper gastrointestinal hemorrhage is greatly influenced by recurrence of bleeding, but it may be possible to identify patients who have a low risk for rebleeding, and can be discharged after a short hospitalization. To examine the effect of an early discharge protocol (length of hospital stay < or =3 days), we conducted a 2-year prospective study in patients with upper gastrointestinal bleeding at low risk for rebleeding, as selected by clinical and endoscopic criteria. METHODS: During the first year of the study, patients were managed according to the standard criteria by any of six surgical teams (control period). During the second year, patients were managed by only one surgical team under the early discharge protocol guidelines (study period). RESULTS: Overall, 488 of 942 (52%) patients were considered as low risk. Early discharge was achieved in 26 of 230 (11%) patients in the control period and in 191 of 258 (74%) in the study period (P <0.001). Age and number of compensated comorbidities did not affect the rate of early discharge. Length of hospital stay was reduced from (mean +/- SD) 6 +/- 2.7 days (control period) to 3 +/- 2.3 days (study period, P <0.001). No differences were observed in rates of rebleeding, need for surgery, readmission or mortality. By contrast, no differences in lengths of stay were observed during that time period among patients admitted with coronary artery disease, colorectal cancer, or acute pancreatitis. CONCLUSION: Most patients with upper gastrointestinal bleeding who are at low risk for rebleeding can be discharged early, leading to important cost savings. PMID- 9753020 TI - A randomized comparison of the safety and efficacy of once-daily gentamicin or thrice-daily gentamicin in combination with ticarcillin-clavulanate. AB - PURPOSE: The primary purpose of the clinical trial was to assess the safety and efficacy of once-a-day compared with three-times-a-day gentamicin in patients with serious infections who had protocol-determined peak serum aminoglycoside concentrations. PATIENTS AND METHODS: A total of 249 hospitalized patients with suspected or proven serious infections were randomized in a 2:2:1 ratio to gentamicin given three times a day with ticarcillin-clavulanate (TC), gentamicin once a day with TC, or ticarcillin-clavulanate (TC) alone. The gentamicin once-a day dosage for patients with estimated creatinine clearance values of > or =80 mL/min was 5.1 mg/kg. With lower creatinine clearance estimates, the mg/kg dosage of gentamicin was decreased, and the dosage intervals (once daily or three times a day) were maintained. Evaluability required documentation of achievement of protocol-defined peak serum gentamicin levels. RESULTS: Of the total 175 evaluable patients, there were no significant differences found between treatment regimens with respect to clinical or microbiologic efficacy. Bedside audiometry proved impractical due to the frequency of altered mental state in ill patients. Based on the traditional increase in serum creatinine values from baseline values, no differences in renal toxicity between the treatment groups was identified. When changes in renal function were reanalyzed based on maintaining, as opposed to worsening, of renal function, preservation of renal function was better in the gentamicin once-a-day patients as opposed to the gentamicin three times-a-day patients, P <0.01. CONCLUSIONS: Gentamicin once a day plus TC, gentamicin three times a day plus TC, and TC alone had similar effects in seriously ill hospitalized patients. The incidence of nephrotoxicity was similar in the three treatment groups. Using a nonvalidated post-hoc analysis, renal function was better preserved in gentamicin once-a-day + TC and TC-only patients as opposed to gentamicin three-times-a-day + TC. PMID- 9753021 TI - Etiology and diagnosis of bilateral leg edema in primary care. AB - PURPOSE: To identify the causes of bilateral leg edema in a primary care setting, and to determine the ability of primary care providers to arrive at the correct diagnosis using the information available at the initial clinical encounter. PATIENTS AND METHODS: Fifty-eight ambulatory adult patients with bilateral leg edema were enrolled at an inner city family practice during a 3-year period. Historical information, physical examination findings, and clinical impressions of primary care providers were compared with the results of laboratory evaluations consisting of echocardiograms, venous duplex ultrasound leg scans, serum albumin levels, and when appropriate, 24-hour urinalyses. RESULTS: Forty five patients (78%) completed the study. The initial clinical impression was venous insufficiency in 32 (71%) patients and congestive heart failure in 8 (18%) patients. In actuality, 15 (33%) patients had a cardiac condition as a cause of their leg edema, and 19 (42%) had pulmonary hypertension. All of the patients with heart disease, and almost all of those with pulmonary hypertension, were age 45 years or older. Only 10 (22%) of the subjects had venous insufficiency. Renal conditions, medication use, and hypoalbuminemia were less common. CONCLUSIONS: Utilizing clinical information only, many patients with cardiopulmonary pathology were incorrectly diagnosed as having more benign conditions, most commonly venous insufficiency. Echocardiographic evaluation, including an estimation of pulmonary artery pressure, may be advisable in many patients with bilateral leg edema, especially if they are at least 45 years old. PMID- 9753022 TI - How you look determines what you find: severity of illness and variation in blood transfusion for hip fracture. AB - PURPOSE: Utilization report cards are commonly used to assess hospitals. However, in practice, they rarely account for differences in patient populations among hospitals. Our study questions were: (1) How does transfusion utilization for hip fracture patients vary among hospitals? (2) What patient characteristics are associated with transfusion and how do those characteristics vary among hospitals? (3) Is the apparent pattern of variation of utilization among hospitals altered by controlling for these patient characteristics? SUBJECTS AND METHODS: We included consecutive hip fracture patients aged 60 years or older who underwent surgical repair between 1982 and 1993 in 19 hospitals from four states, excluding those who refused blood transfusion, had multiple trauma, metastatic cancer, multiple myeloma, an above the knee amputation, or were paraplegic or quadriplegic. The outcome of interest was postoperative blood transfusion. "Trigger hemoglobin" was the lowest hemoglobin recorded before transfusion or recorded at any time during the week before or after surgery for patients who were not transfused. RESULTS: There was considerable variation in transfusion among hospitals postoperatively (range 31.2% to 54.0%, P = 0.001). Trigger hemoglobin also varied considerably among hospitals. In unadjusted analyses, four of nine teaching and two of nine nonteaching hospitals had postoperative transfusion rates significantly higher than the reference (teaching) hospital, while one nonteaching hospital had a lower rate. In an analysis controlling for trigger hemoglobin and multiple clinical variables, one of nine teaching and four of nine nonteaching hospitals had rates higher than the reference hospital, while four teaching hospitals and one nonteaching hospital had lower rates. CONCLUSIONS: The apparent pattern of variation of transfusion among hospitals varies according to how one adjusts for relevant patient characteristics. Utilization report cards that fail to adjust for these characteristics may be misleading. PMID- 9753023 TI - Myocardial fibrin deposits in the first month after transplantation predict subsequent coronary artery disease and graft failure in cardiac allograft recipients. AB - PURPOSE: To determine whether fibrin deposition during the first month following cardiac transplantation predicts development of coronary artery disease and graft failure in cardiac allograft recipients. PATIENTS AND METHODS: We prospectively studied 121 consecutive adult patients who received cardiac transplants between 1988 and 1995. Serial endomyocardial biopsies obtained during the first month posttransplant (2.3 + 0.6 biopsies/patient) were studied immunohistochemically for fibrin deposits. Patients were followed up with annual angiograms (3.2 + 1.7/patient) evaluated with side-by-side comparisons for the presence and progression of coronary artery disease. RESULTS: All pretransplant biopsies were fibrin-negative; 60 allografts (50%) remained without fibrin, and 61 (50%) contained fibrin during the first posttransplant month. Of allografts with fibrin, 72% developed coronary artery disease, while 27% of allografts without fibrin developed the disease (P <0.001). Coronary artery disease was progressive in 61% of allografts with fibrin, and in 25% of allografts without fibrin (P <0.001). Graft failure was more frequent and time-to-graft-failure occurred earlier in patients whose allografts had fibrin during the first month after transplantation (P <0.001). CONCLUSIONS: Fibrin in biopsies during the first month after transplantation identifies patients at high risk for developing coronary artery disease or graft failure, thereby allowing the opportunity to initiate preventive procedures. PMID- 9753024 TI - Psychiatric illness in patients with end-stage renal disease. AB - PURPOSE: We sought to determine the prevalence of psychiatric illness in hospitalized patients with end-stage renal disease. We also examined the association between end-stage renal disease treatment modality and risk of hospitalization with a diagnosis of a mental disorder, and compared rates of hospitalization with a diagnosis of psychiatric illness in renal failure patients to patients with other chronic medical illnesses. SUBJECTS AND METHODS: We performed a cohort study of all Medicare-enrolled dialysis patients in 1993. Risk of hospitalization with a diagnosis of a mental disorder among renal failure patients was compared with Medicare patients with diabetes mellitus, ischemic heart disease, cerebrovascular disease, and peptic ulcer disease. RESULTS: Almost 9% of all dialysis patients were hospitalized with a mental disorder. Men, African-Americans, and younger patients were more likely to be hospitalized with a mental disorder. The adjusted risk of hospitalization for peritoneal dialysis patients was lower compared with hemodialysis patients for any mental disorder, depression, and alcohol and drug use. Hospitalization with mental disorders was 1.5 to 3.0 times higher for renal failure patients compared with other chronically ill patients. CONCLUSIONS: Hospitalization with a psychiatric illness is common among the US end-stage renal disease population. Depression, dementia and drug-related disorders were especially common. The coexistence of psychiatric illness in patients with renal failure who require specialized medical regimens represents a challenge to nephrologists in diagnosis and treatment. Disparities between hospitalization rates of psychiatric illnesses among end-stage renal disease patients compared with other chronically ill populations warrant further research. PMID- 9753025 TI - Outcomes, preferences for resuscitation, and physician-patient communication among patients with metastatic colorectal cancer. SUPPORT Investigators. Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatments. AB - PURPOSE: To describe characteristics, outcomes, and decision making in patients with colorectal cancer metastatic to the liver, and to examine the relationship of doctor-patient communication with patient understanding of prognosis and physician understanding of patients' treatment preferences. PATIENTS AND METHODS: The Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatments (SUPPORT) was a prospective cohort study conducted at five teaching hospitals in the United States between 1989 and 1994. Participants in this study were hospitalized patients 18 years of age or older with known liver metastases who had been diagnosed with colorectal cancer at least 1 month earlier. Data were collected by patient interview and chart review at study entry; patients were interviewed again at 2 and 6 months. Data collected by physician interview included estimates of survival and impressions of patients' preferences for cardiopulmonary resuscitation (CPR). Patients and physicians were also asked about discussions about prognosis and resuscitation preferences. RESULTS: We studied 520 patients with metastatic colorectal cancer (median age 64, 56% male, 80% white, 2-month survival 78%, 6-month survival 56%). Quality of life (62% "good" to "excellent") and functional status (median number of disabilities = 0) were high at study entry and remained so among interviewed survivors at 2 and 6 months. Of 339 patients with available information, 212 (63%) of 339 wanted CPR in the event of a cardiopulmonary arrest. Factors independently associated with preference for resuscitation included younger age, better quality of life, absence of lung metastases, and greater patient estimate of 2-month prognosis. Of the patients who preferred not to receive CPR, less than half had a do-not resuscitate note or order written. Patients' self-assessed prognoses were less accurate than those of their physicians. Physicians incorrectly identified patient CPR preferences in 30% of cases. Neither patient prognostication nor physician understanding of preferences were significantly better when discussions were reported between doctors and patients. CONCLUSIONS: A majority of patients with colorectal cancer have preferences regarding end of life care. The substantial misunderstanding between patients and their physicians about prognosis and treatment preferences appears not to be improved by direct communication. Future research focused on enhancing the effectiveness of communication between patients and physicians about end of life issues is needed. PMID- 9753026 TI - Antihypertensive drugs and the risk of gastrointestinal bleeding. AB - PURPOSE: Calcium channel blockers have been reported to increase the risk of gastrointestinal bleeding. We tested this hypothesis, and also assessed whether beta blockers decrease this risk. SUBJECTS AND METHODS: A nested case-control design within a population-based cohort of all 34,074 new users of beta blockers, angiotensin-converting enzyme (ACE) inhibitors, or calcium channel blockers in Saskatchewan, from 1990 to 1993 and followed up to March 1995, was used. We identified all 311 subjects hospitalized because of gastrointestinal bleeding during this period, each of whom was matched to 10 randomly selected controls. RESULTS: The rate of hospitalization for gastrointestinal bleeding was 3.0 per 1,000 per year. The adjusted rate ratio of gastrointestinal bleeding for current use of calcium channel blockers was 1.1 (95% confidence interval [CI] 0.8 to 1.4) and 0.66 (95% CI 0.44 to 0.98) for beta blockers compared with no current use of anti-hypertensive drugs. The adjusted rate ratio for ACE inhibitor use was 1.0 (95% CI 0.7 to 1.3) while that for diuretic use was 1.4 (95% CI 1.0 to 2.0). CONCLUSIONS: The use of calcium channel blockers does not appear to increase the risk of gastrointestinal bleeding in the first five years of treatment, while beta blockers may prevent this adverse event. The unexpected elevated risk associated with the use of diuretics needs to be investigated further. PMID- 9753027 TI - The efficacy of silver alloy-coated urinary catheters in preventing urinary tract infection: a meta-analysis. AB - PURPOSE: Indwelling urinary catheters are implicated in most cases of nosocomial urinary tract infection. Silver-coating of catheters may reduce the risk of these infections; however, trials have provided mixed results. We performed a meta analysis to estimate the effectiveness of silver-coated urinary catheters. SUBJECTS AND METHODS: Published or unpublished articles were sought using MEDLINE, reference review, and correspondence with original authors, catheter manufacturers, and experts. Trials using silver-coated urinary catheters in the treatment group and uncoated urinary catheters in the control group were included. Bacteriuria, as evaluated by urine culture, was the outcome variable used to indicate urinary tract infection. Summary odds ratios (OR) and 95% confidence intervals (CI) were calculated using Mantel-Haenszel methods with a fixed-effects model. RESULTS: Of 117 reports retrieved, eight trials with a total of 2,355 patients satisfied inclusion criteria. The summary OR for urinary tract infection was 0.59 (95% CI, 0.42 to 0.84) indicating a significant benefit in the patients receiving silver-coated catheters. A test of heterogeneity, however, indicated that the odds ratios varied significantly among studies. Silver alloy catheters (OR = 0.24; 95% CI, 0.11 to 0.52) were significantly more protective against bacteriuria than silver oxide catheters (OR = 0.79; 95% CI, 0.56 to 1.10). CONCLUSIONS: This meta-analysis clarifies discrepant results among trials of silver-coated urinary catheters by revealing that silver alloy catheters are significantly more effective in preventing urinary tract infections than are silver oxide catheters. Though silver alloy urinary catheters cost about $6 more than standard urinary catheters, they may be worth the extra cost since catheter related infection is a common cause of nosocomial infection and bacteremia. PMID- 9753028 TI - Diseases of receptors: introductory comments. PMID- 9753029 TI - Receptors: structure and function. PMID- 9753030 TI - Acute renal failure in a marathon runner: role of glomerular bleeding in tubular injury. PMID- 9753031 TI - Cardiac tamponade in hereditary hemorrhagic telangiectasia. PMID- 9753032 TI - Aminoglycoside dosing: timing is of the essence. PMID- 9753033 TI - Criteria for evaluating disease response and progression in patients with multiple myeloma treated by high-dose therapy and haemopoietic stem cell transplantation. Myeloma Subcommittee of the EBMT. European Group for Blood and Marrow Transplant. PMID- 9753034 TI - Differentiation therapy of myelodysplastic syndromes: fact or fiction? PMID- 9753035 TI - Functional, phenotypic and molecular characterization of cytokine low-responding circulating CD34+ haemopoietic progenitors. AB - Circulating CD34+ cell populations characterized by a low rate (up to five) or high rate (more than five) of cell divisions were isolated from 8 d cultures in the presence of stem cell factor (SCF), interleukin-3 (IL-3), granulocyte macrophage colony stimulating factor (GM-CSF), granulocyte colony stimulating factor (G-CSF), erythropoietin (EPO), Flt3 ligand and Peg-rHu megakaryocyte growth and development factor (Peg-rHuMGDF) using the fluorescent dye 5,6 carboxyfluorescein diacetate succinimidyl ester (CFDA-SE) and flow cytometric cell sorting. Phenotypic characterization of cells which had experienced up to five divisions (CFDA-SEbright) showed a similar surface antigen expression to starting, freshly isolated CD34+ cells. Conversely, cells which experienced more than five divisions (CFDA-SEdim) showed a differentiating behaviour, down regulating CD34 antigen and acquiring differentiation markers. CFDA-SEbright cells were significantly enriched in CD105 (endoglin) positive precursors as compared to both freshly isolated CD34+ and CFDA-SEdim cells. Functional analysis indicated that CFDA-SEbright had a 3-fold and 10-fold greater cumulative cloning efficiency as compared to freshly isolated CD34+ cells and CFDA-SEdim cells, respectively. CFDA-SEbright cells retained the vast majority of LTC-IC and showed a LTC-IC frequency 2.8-fold higher than that found in freshly isolated CD34+ cells. RT-PCR and Western blot analyses showed significantly higher bcl-2 RNA and protein levels in CFDA-SEbright cells as compared to freshly isolated CD34+ and CFDA-SEdim cells. This study indicates that cytokine low-responding circulating CD34+ cells (CFDA-SEbright cells) represent a functionally, phenotypically and molecularly distinct multipotent progenitor population with biological properties associated with primitive precursors. PMID- 9753036 TI - Elevated SCF levels in the serum of patients with chronic renal failure. AB - Serum stem cell factor (SCF) and soluble KIT (sKIT) levels were estimated in patients with chronic renal failure (CRF) and anaemia, and compared with clinical parameters of blood cells and renal function. Serum SCF levels in CRF patients were 5-fold higher than those in healthy controls. However, serum sKIT levels in haemodialysis (HD)-CRF patients were only slightly higher than those of healthy controls. In untreated CRF patients and healthy controls, serum SCF levels were significantly correlated with blood urea nitrogen (BUN), creatinine. haemoglobin, red blood cell (RBC) count and sKIT. In untreated CRF patients, serum SCF levels were significantly correlated with BUN, creatinine, and sKIT. These results suggest that serum SCF levels increased with the deterioration of renal function and might be related to erythropoiesis. PMID- 9753037 TI - Decreased activity of the multidrug resistance P-glycoprotein in acquired aplastic anaemia: possible pathophysiologic implications. AB - To address a possible impairment of multidrug resistance mechanisms in acquired aplastic anaemia (AA), the functions of P-glycoprotein (P-gp) and multidrug resistance-associated protein (MRP) were respectively assessed by rhodamine 123 (Rh123) and daunorubicin (DNR) efflux in peripheral blood lymphocytes from AA patients. The proportion of Rh123-effluxing T cells was significantly decreased in AA, relative to controls. Interestingly, these changes were also present in patients with AA in remission. Conversely, Rh123 efflux in B and natural killer (NK) cells and DNR efflux in peripheral blood lymphocytes were unchanged. These data indicated that P-gp activity was decreased in AA not only during the development of the disease, but also after remission, introducing a new concept on the pathophysiology of AA by suggesting that it may contribute to drug-induced injury to haemopoietic cells in some cases of AA, by increasing the proportion of susceptible cells. PMID- 9753038 TI - Chromosomal breakage analysis in dyskeratosis congenita peripheral blood lymphocytes. AB - Dyskeratosis congenita (DC) is a rare inherited disorder characterized by reticulate skin pigmentation, nail dystrophy and mucosal leucoplakia. Bone marrow failure occurs in the majority of cases and there is a predisposition to malignancy. Following conflicting reports of increased spontaneous and induced chromosomal breakage in DC lymphocytes, we examined chromosomal breakage with and without clastogen treatment in 10 DC patients from six different families. Peripheral blood cultures were stimulated with phytohaemagglutinin and treated with three clastogenic agents and gamma-irradiation. There was no significant difference in the chromosomal breakage in DC lymphocytes with or without exposure to bleomycin, DEB, MMC or gamma-irradiation. DC can therefore be distinguished from Fanconi's anaemia in which lymphocytes show increased spontaneous and clastogen-induced chromosomal breakage. PMID- 9753039 TI - Modulating the oxygen affinity of human fetal haemoglobin with synthetic allosteric modulators. AB - Improving the delivery of oxygen to the tissues by decreasing the oxygen affinity of haemoglobin has been a major aim of several laboratories over recent years because this may reduce the consequences of anaemia and/or improve tissue oxygenation in cases of decreased blood perfusion. Within the same context, lowering the oxygen affinity may prove valuable in the application of native or recombinant haemoglobin solutions as a blood substitute. The shift of the oxygen equilibrium curve to the right is obtained by various modulators. Among them, the bezafibrate derivatives are considered as a most interesting group. These principles are of the utmost importance in thalassaemia and other haemoglobinopathies where the beneficial effects of the compensatory synthesis of fetal haemoglobin are diminished by the increased oxygen affinity of this pigment. In this paper we present the results of a study initiated to determine whether a potent oxygen affinity modifier, RSR-4, could satisfactorily decrease the oxygen affinity of fetal haemoglobin, thus improving tissue oxygenation. The experiments were carried out on whole blood and on purified haemoglobin solutions and showed that the effector markedly decreased the oxygen affinity of HbF (from 18.7 to 3.73 mmHg in whole blood). At the same time the cooperativity index (n50) and the oxygen saturation levels remained within normal limits under the conditions of the main experiment. These observations have important implications for the potential application of oxygen affinity modifiers in vivo. PMID- 9753040 TI - Prevalence of hereditary haemochromatosis in premature atherosclerotic vascular disease. AB - It has been proposed that iron accumulation may contribute to atherogenesis by increasing free radical formation and oxidative stress. Epidemiological studies in which the association of iron status with atherosclerosis was assessed raised conflicting results. To test whether genetic haemochromatosis is associated with increased atherosclerosis, we determined the prevalence of two mutations in the HFE gene related to haemochromatosis (845G-->A; Cys282Tyr. and 187 C-->G, His63Asp) in 265 consecutive patients with premature (<50 years of age) angiographically-proven atherosclerotic disease (coronary and/or peripheral), and in 272 healthy controls. PCR amplification followed by RsaI (Cys282Tyr analysis) and BclI (His63Asp analysis) restriction digestion was employed to define the genotypes. The mutant Cys282Tyr allele had a frequency of 0.07 among controls and 0.04 among patients (carrier frequency of 14.0% and 8.3%, respectively). The frequency of the His63Asp mutant allele was 0.14 (28.6% of carriers) in controls and 0.11 (22.2% of carriers) in patients. Five of 265 patients (1.1%) and 9/272 controls (3.3%) were compound heterozygotes. In conclusion, a lower prevalence of the Cys 282Tyr mutation and a similar frequency of the His63Asp mutation was observed in patients with atherosclerotic disease in comparison with normal controls. These findings do not support an association between haemochromatosis and atherogenesis. PMID- 9753041 TI - Is there a link between African iron overload and the described mutations of the hereditary haemochromatosis gene? AB - Over 80%, of Caucasian patients with hereditary haemochromatosis are homozygotes for a C282Y mutation in the HFE gene on chromosome 6. Recent evidence suggests that a genetic factor may also be involved in the pathogenesis of African iron overload, although the locus has not been described. PCR analysis of DNA from 25 southern Africans, identified by segregation analysis as having a high probability of carrying the putative African iron-loading gene, failed to identify any subjects with the C282Y mutation. The possible genetic defect in African iron overload appears to be different from that described in most cases of hereditary haemochromatosis in Caucasians. PMID- 9753042 TI - Aetiology of severe iron overload in a family with hereditary haemolytic anaemia. AB - Severe iron overload is a reported complication of certain erythroid disorders which are characterized by increased erythropoietic activity. Proposed mechanisms include enhancement of iron absorption secondary to increased erythroid activity and coexistent heterozygosity or homozygosity for haemochromatosis. We performed PCR-based analysis for the haemochromatosis-related HFE C282Y mutation in an extended family with inherited haemolytic anaemia in which several members exhibited iron overload. The results demonstrated iron overload was associated with homozygosity but not heterozygosity for this mutation. Such an association may also exist in other erythroid disorders in which iron overload has been reported. PMID- 9753043 TI - Successful management of concurrent congenital dyserythropoietic anaemia and autoimmune haemolytic anaemia with splenectomy. AB - This first known case of concurrent congenital dyserythropoietic anaemia (CDA) and autoimmune haemolytic anaemia (AIHA) which occurred in a hispanic male and spanned 6 years from the age of 2. Light and electron microscopy of bone marrow erythroblasts and immunophenotyping confirmed CDA; serum/eluate warm autoantibodies and positive direct antiglobulin tests (DAT) associated with severe, episodic anaemias established AIHA. Cytogenetic analysis of bone marrow cells and peripheral blood lymphocytes ascertained sex chromosome aneuploidy (48 XY,+ Y,+ Y). Recurrent, life-threatening episodes of transfusion-dependent anaemia refractory to steroids and intravenous immune globulin, were put into stable remission at age 8 years when splenectomy successfully managed both disorders. PMID- 9753044 TI - Low CD4 counts rather than superantigenic-like effects account for differences in expressed T-cell receptor (TCR) repertoires between HIV-1 seropositive long-term non-progressors and individuals with progressive disease. AB - Analysis of HIV-infected individuals who have stable CD4 counts many years after seroconversion may provide an insight as to how the host's immune system can successfully control HIV infection. In this study we analysed the T-cell receptor (TCR) Vbeta repertoire in 13 HIV+ individuals, seven of whom were classed as long term non-progressors (LTNP), using a technique which couples anchor PCR (AnPCR) amplification of beta-chain cDNA to differential probe hybridization with non radioactively labelled Vbeta family specific oligonucleotide probes. There were no significant differences in the expressed TCR repertoires between the LTNP group and the other HIV-infected individuals. However, there was a statistically significant inverse correlation between CD4 count and the number of Vbeta family specific perturbations in the recent seroconverters (SC) and those with progressive infection (PI), consistent with other shared features of clinical disease progression (Th1/Th2 switch and high frequency of viral isolation). We conclude that long-term clinical non-progression in HIV-1 infection is not associated with the loss or expansion of a particular Vbeta family; instead, low CD4 count in the PI and SC individuals was correlated with increased number of Vbeta family-specific perturbations relative to the LTNP group and that it is hence unlikely that HIV encodes a superantigen. PMID- 9753045 TI - Effect of granulocyte-macrophage colony-stimulating factor treatment on phenotype, cytokine release and cytotoxicity of circulating blood monocytes and monocyte-derived macrophages. AB - We studied phenotype, function and differentiation of mononuclear phagocytes in 11 cancer patients treated subcutaneously with 10O microg/kg recombinant human (rhu) GM-CSF for 7 d. The rhuGM-CSF treatment induced (1) a 5.9-fold increase in the number of blood monocytes (MO), (2) a decrease of CD14bright/CD16bright cells with a diminution of the mean fluorescence intensity (MFI) of CD14, and (3) a decrease of MO cellular cytotoxicity. In patients' sera, tumour necrosis factor (TNF)-alpha, interleukin (IL)-10, IL-12, neopterin, macrophage-colony-stimulating factor (M-CSF), and IL-1 receptor antagonist (IL-1RA) increased, whereas GM-CSF and granulocyte-colony-stimulating factor (G-CSF) decreased after an initial peak. In whole blood samples the lipopolysaccharide (LPS)-stimulated release of TNF-alpha, IL-6 and IL-1RA increased initially, whereas IL-1beta, IL-10 and IL-12 decreased. During differentiation from MO to macrophages (MAC), interferon (IFN) gamma-stimulated tumour cytotoxicity increased, but both MO and MAC were less cytotoxic upon rhuGM-CSF treatment. The differentiation-associated increase of LPS-induced TNF-alpha, IL-1RA and IL-10 secretion was reduced by the rhuGM-CSF treatment, and the expression of CD14 on MAC as well as the proportion of CD14+/CD16+, CD14+/MAX.1+ and CD14+/CD71+ cells in 7-d cultured MAC declined. We interpret these findings as (1) an increase of immature MO upon rhuGM-CSF therapy, (2) a priming effect on the LPS-induced proinflammatory cytokine repertoire of MO, and (3) an impact of rhuGM-CSF on the capacity of MO to differentiate to MAC in vitro. PMID- 9753046 TI - Distribution of prostaglandin IP and EP receptor subtypes and isoforms in platelets and human umbilical artery smooth muscle cells. AB - Prostaglandins act through specific receptors to stimulate cyclic AMP formation which inhibits platelet activation and relaxes vascular smooth muscle. We have used RT-PCR combined with Southern blot analysis to determine the subtypes of prostaglandin receptor on platelets. Platelets expressed the EP4 rather than the EP2 prostaglandin EP receptor subtype, whereas vascular smooth muscle cells predominantly expressed the EP2 receptor. The IP receptor, which binds prostacyclin and couples to stimulation of adenylyl cyclase, and three isoforms of the inhibitory EP3 receptor were equally expressed in platelets, HEL cells and umbilical artery smooth muscle cells. The EP3-II isoform showed variation in level of expression among the three cell types. As a positive control for the presence of platelet RNA, PCR was performed using primers specific for the alpha chain of the platelet membrane glycoprotein Ib. As a negative control for the absence of T and B cell contamination in the platelet RNA, PCR was performed using primers specific for the cell specific cluster determinants CD2 (a T-cell marker) and CD20 (a B-cell marker). The finding that platelets express both stimulatory and inhibitory prostaglandin receptors provides confirmation of a homeostatic model of regulation of platelet adenylyl cyclase previously proposed. PMID- 9753047 TI - Flow cytometric analysis of platelet activation by different collagen types present in the vessel wall. AB - The interaction of platelets with collagens of the vessel wall is a critical event in primary haemostasis. Although numerous studies have examined the ability of various collagen types to support platelet adhesion, little is known concerning the relative ability of different collagens to elicit specific activation markers in platelets. In this report, flow cytometric analysis has been utilized to evaluate the ability of various native collagen types to elicit secondary activation events in human platelets. Collagen types I, III, V and VI induced alpha-granule secretion and up-regulation of cell surface glycoprotein (GP) IIb/IIIa. In contrast, collagen type IV did not elicit these responses in the concentration ranges examined. Dose-response curves for alpha-granule secretion induced by the various collagen types indicated that human type III and human type I collagens were less effective than human type V, human type VI and calf skin type I. In addition, the ability of these various collagens to activate GPIIb/IIIa to its ligand binding conformation was even more heterogenous with only human type VI and calf skin type I readily promoting this transition. These data demonstrate that flow cytometric analysis of collagen-induced platelet activation is feasible and that collagen-mediated alpha-granule secretion and membrane glycoprotein redistribution in human platelets are separate events from activation of GPIIb/IIIa. PMID- 9753048 TI - Protein Z in healthy human individuals and in patients with a bleeding tendency. AB - Protein Z is a vitamin-K-dependent plasma glycoprotein; its physiological function is not clear. Low protein Z levels were found in patients with otherwise unexplained bleeding disorders. It was therefore suggested that low protein Z levels might be associated with a bleeding diathesis. In the present study we measured protein Z levels in plasma samples of 48 patients with a suspected bleeding disorder and in plasma samples of 200 healthy adult individuals. We found protein Z to have a wide normal range in healthy men and women. Significantly lower protein Z levels were observed in women compared to men, whereas no correlation was found with age or other vitamin-K-dependent coagulation proteins. None of the 48 patients with a bleeding disorder had a protein Z level below the normal range. However, protein Z was significantly lower in the group of male patients with a bleeding history as compared to healthy men. In conclusion, our data indicate that low-normal protein Z levels are not associated with a bleeding tendency. However, it remains to be determined whether a low protein Z level is a weak cofactor associated with an increased bleeding tendency and whether decreased or absent protein Z (conditions not detected in our patients) might constitute a haemorrhagic diathesis. PMID- 9753049 TI - A prospective clinical trial of implantable central venous access in children with haemophilia. AB - To assess the risks associated with the use of central venous ports in children with haemophilia, 15 HIV-negative patients were prospectively evaluated. Port insertion was required for immune tolerance in two inhibitor patients and continuous prophylaxis in 13 patients with severe factor VIII deficiency, for whom surgery was covered with recombinant factor VIII (rFVIII), then given daily at home until day 6. One inhibitor patient (titre 7BU/ml) received high-dose rFVIII by continuous infusion until day 3, followed by an immune tolerance treatment scheme; the other (titre 12 BU/ml) was given recombinant activated factor VII by continuous infusion until day 7. After training on the use of the port, all patients continued their infusion programme at home. All ports remained in place for a median period of 413d (range 125-509). The median number of entries into the port was 184 (range 53-567). Port-site haematoma and infection occurred in one patient on day 7 when an inhibitor became detectable (titre 12 BU/ml). An infectious complication occurred in another patient after 310d. The port infection rate was 0-42 per 1000 patient-days (0.33 per 1000 entries into the port). This protocol for port placement with short hospitalization appears feasible and safe. PMID- 9753050 TI - Low-dose aspirin does not lower in vivo platelet activation in healthy smokers. AB - Smoking causes atherosclerosis, and smokers have increased thromboxane (TXA2) formation. As aspirin inhibits TXA2 production we speculated that smokers would preferentially profit from inhibition of the TXA2 pathway by aspirin. Increased expression of P-selectin, a constituent of the alpha-granules of platelets, and increased levels of circulating (c)P-selectin in plasma are markers for platelet activation. The aim of this study was to compare P-selectin expression on platelets between smokers and nonsmokers, and to compare with placebo the effect of 2 weeks administration of 100 mg/d aspirin on platelet activation in smokers. Smokers exhibited higher P-selectin expression on platelets than non-smokers (2.7+/-1.8% v 1.6+/-0.6%, P=0.018), thus confirming increased platelet activation. Aspirin did not reduce platelet activation as demonstrated by unchanged P-selectin expression on platelets and cP-selectin plasma levels. PMID- 9753051 TI - Effect of thrombopoietin on proliferation of blasts from CD7-positive acute myelogenous leukaemia. AB - We investigated the effect of thrombopoietin (TPO) on the growth of leukaemic blasts from 30 acute myelogenous leukaemia (AML) patients according to the surface expression of CD7 and CD34: 10 patients were CD7 positive (CD7+), nine were CD7 negative/CD34+ (CD7-/CD34+) and the remaining 11 were CD7-/CD34-. Significant growth response of leukaemic blasts to TPO was observed in 10/10 CD7+, 5/9 CD7-/CD34+ and 2/11 CD7-/CD34- AML cases using 3H-thymidine incorporation. Synergistic stimulatory effects of TPO with stem cell factor (SCF), interleukin-3 (IL-3), granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor were observed in both TPO responding cases (9/17) and TPO-non-responding cases (8/13). In a leukaemic blast colony assay. significant growth response to TPO was observed in 5/6 CD7+ and 4/17 CD7-AML cases examined. However, the effect of TPO on the growth of CD7+ leukaemic blasts was not so potent as that of IL-3 and SCF, both of which support the proliferation of primitive haemopoietic progenitors. Expression of c-mpl (TPO receptor) was significantly higher in CD7+ AML cases than in CD7- cases, suggesting a relationship between expression of c-mpl and proliferative response to TPO. These data indicate that CD7+ leukaemic blasts express functional TPO receptors and proliferate in response to TPO. These observations also imply that CD7 expression on AML blasts may indicate involvement of leukaemic progenitors at an early stage of multipotent haemopoietic stem cells. PMID- 9753052 TI - Absence of Bruton's tyrosine kinase (Btk) mutations in patients with acute myeloid leukaemia. AB - Bruton's tyrosine kinase (Btk) is a non-receptor protein tyrosine kinase (PTK) that is expressed in all haemopoietic lineages except mature T cells and plasma cells. Despite the broad range of expression. mutations that inactivate this molecule affect primarily the development of the B-cell lineage. As a PTK, Btk could potentially be involved directly or indirectly in the processes that relate to the malignant transformation of all the cell lineages where this molecule is expressed. Previous studies have failed to demonstrate mutations in patients with B-cell origin acute lymphoblastic leukaemia (ALL). We have utilized a recently developed method that enables the rapid and convenient detection of mutations at the cDNA level, namely, the non-isotopic RNase cleavage assay (NIRCA) to analyse Btk sequences from 27 patients with different types of acute myeloid leukaemia (AML). The only alteration that we observed was a polymorphism at position 2031. This polymorphism has already been seen in previous studies. Furthermore, using the same methodology, we identified the Btk mutations in six XLA (X-linked agammaglobulinaemia) patients. Our results, although they do not exclude the involvement of Btk mutations in the development or progression of some type of AML, nevertheless suggest that such mutations do not constitute a major co-factor in the development of myeloid malignancies. PMID- 9753053 TI - Establishment of a novel human myeloid leukaemia cell line (FKH-1) with t(6;9)(p23;q34) and the expression of dek-can chimaeric transcript. AB - Translocation t(6:9)(p23;q34), resulting in a dek-can gene fusion, is a recurrent chromosomal abnormality mainly associated with specific subtypes of acute myeloid leukaemia (AML) and myelodysplastic syndrome (MDS). Patients with this type of chromosomal change are usually young and their prognosis is poor. The role of fusion protein generated from dek-can chimaeric transcript on the leukaemogenesis oft(6;9) AML or MDS is as yet unknown. We have established the first permanent cell line (FKH-1) with t(6;9). derived from the peripheral blood of a patient with t(6:9) AML transformed from Philadelphia chromosome (Ph1)-negative chronic myelocytic leukaemia (CML). The FKH-1 expressed myelomonocytic markers and dek can chimaeric transcript. In the presence of 10 ng/ml recombinant human granulocyte colony-stimulating factor (G-CSF), the cells doubled every 54 h and showed multilineage myeloid differentiation, resulting in heterogenous morphologies such as macrophages, basophils, eosinophils and neutrophils. Thus, this cell line may be derived from a pluripotent myeloid stem cell and should be a useful tool for biomolecular studies on the pathogenesis of t(6;9) myeloid malignancies which have rarely been investigated because of the lack of continuously proliferating cells. PMID- 9753054 TI - The expression of co-stimulatory molecules and their relationship to the prognosis of human acute myeloid leukaemia: poor prognosis of B7-2-positive leukaemia. AB - We examined the expression of co-stimulatory molecules on leukaemic cells of 52 adult patients with acute myeloid leukaemia (AML) (34 men and 18 women) and analysed the relationship between these expressions and the patient's prognosis. B7-1 was not expressed in any of the 23 patients investigated, whereas B7-2 was expressed in 26/52 patients (50.0%). B7-2 was expressed in all AML patients with monocytic morphology (M4 or M5) and in 16/42 cases without monocytic morphology. CD54 was expressed in 28/ 37 patients examined (75.7%), and CD58 was expressed in all of the AML patients except one (M 7). The overall survival of the 26 B7-2 positive leukaemia patients (1-24 months, median survival 11.5 months) was significantly shorter than that of the 26 B7-2-negative leukaemia patients (1-71+ months, median 35.1 months) (P=0.0080). In addition, the B7-2-positive patients exhibited significantly shorter disease-free survival periods compared to the B7 2-negative patients (P=0.021). There was no significant difference in age, sex, haematological data and complete remission rate between the B7-2-positive and B7 2-negative patients. Our results indicated that B7-2 is one of the most crucial factors in the prognosis of adult acute leukaemia and can be expected to have an important role in tumour immunity. PMID- 9753055 TI - Shift from Rh-positive to Rh-negative phenotype caused by a somatic mutation within the RHD gene in a patient with chronic myelocytic leukaemia. AB - We report a female patient whose Rh phenotype shifted from RhD-positive to RhD negative over a 3-year period (1991-94), during which time she was treated with mastectomy (1992) and local irradiation for a low-grade recurrent breast cancer. She was diagnosed with chronic myeloid leukaemia in 1994, and has since then received chemotherapy. The patient was repeatedly typed as O, RhD-positive between 1965 and 1991 and was repeatedly found RhD-negative after 1994. Bcr-Abl transcripts typical of Ph1 chromosome were detected. Molecular analysis indicated that the patient was heterozygous at the RH locus, carrying one haplotype in which the RHD gene exhibited a single nucleotide deletion (G600) resulting in a frameshift and premature stop codon, and a normal RHCE gene (allele Ce). The second haplotype contained only the RHCE gene (allele ce) and was normal. Further analysis carried out on total leucocytes, purified neutrophils, EBV lymphoblastoid cell line and cultured erythroblasts indicated that the G600 deletion was restricted to the myeloid lineage. No modification of other blood group antigens could be detected. These findings suggest a somatic mutation which most probably occurred in a stem cell common to the myeloid lineage. PMID- 9753056 TI - BCR-ABL-positive progenitors in chronic myeloid leukaemia patients in complete cytogenetic remission after treatment with interferon-alpha. AB - To determine the source of residual disease detected in patients with chronic myeloid leukaemia (CML) in complete cytogenetic remission (n=8) after treatment with interferon-alpha (IFN-alpha), we have tested CFU-GM colonies grown from bone marrow mononuclear cells or from plastic-adherent (Pdelta) cells for BCR-ABL mRNA using a nested multiplex RT-PCR. We compared our results with those obtained by analysis of colonies from newly diagnosed patients (n=4) and patients achieving no cytogenetic response (n=1) or incomplete cytogenetic response to treatment with IFN-alpha (n=5). A total of 1239 informative colonies were analysed. A small proportion of BCR-ABL-positive colonies was detected in all eight patients in complete cytogenetic remission, suggesting the persistence of leukaemia that could potentially lead to relapse. The overall proportion of BCR-ABL-positive colonies in patients achieving a cytogenetic response to IFN-alpha correlated with the levels of BCR-ABL transcripts detected in the peripheral blood by competitive RT-PCR (P=0.004). We conclude that residual disease detected in the peripheral blood of complete cytogenetic responders to IFN-alpha is at least partly derived from clonogenic myeloid cells. It is probable that the leukaemia clone in CML is only very rarely or never entirely eradicated by treatment with IFN-alpha. PMID- 9753057 TI - Lack of association between childhood common acute lymphoblastic leukaemia and an HLA-C locus dimorphism influencing the specificity of natural killer cells. AB - Previous serological studies documenting an association between acute lymphoblastic leukaemia (ALL) and HLA-Cw antigens suggested that the HLA-C locus might influence susceptibility to ALL. However, associations with more than one Cw antigen suggest that polymorphic variants shared by more than Cw allele could be involved. Recent studies have shown that the HLA-C locus encodes two ligands (NK1 and NK2) recognized by receptors on natural killer (NK) cells. HLA-Cw alleles encoding these ligands are dimorphic, dependent on whether they encode one or other NK ligand. To determine whether susceptibility to the common (CD10+) form of childhood ALL (c-ALL) is associated with NK1 or NK2, we carried out a molecular analysis of 94 childhood c-ALL patients and 136 infant controls. We found no difference in the frequency of NK1 and NK2 alleles, phenotypes or genotypes between the patients and controls, suggesting that this does not explain the role of the HLA-C locus in susceptibility to childhood c-ALL. PMID- 9753058 TI - Prolonged but reversible neutrophil dysfunctions differentially sensitive to granulocyte colony-stimulating factor in children with acute lymphoblastic leukaemia. AB - Treatment of average-risk acute lymphoblastic leukaemia (ALL) in children consists of 6 months of intensive chemotherapy followed by 18 months of maintenance therapy. Polymorphonuclear leucocyte (PMN) functions from children with ALL were studied in order to evaluate and compare the toxicity of the initial intensive treatment with the toxicity of the subsequent less intensive maintenance treatment. H2O2 and O2- production, evaluated by chemiluminescence, were significantly decreased during the intensive period but returned to normal values when maintenance therapy began. In contrast, bactericidal activity against Gram-positive and Gram-negative microorganisms remained at low levels throughout the treatment but returned to normal values in patients off chemotherapy. PMN from patients on maintenance therapy exhibited an excess of morphological changes associated with apoptosis. This was confirmed by standard two-colour flow cytometry which revealed an increase in the number of hypodiploid cells, and increased expression of membrane phosphatidylserine together with a drastic reduction in the expression of the Fcgamma receptor IIIB (CD16). These defective PMN were differentially sensitive to the effects of granulocyte colony stimulating factor (G-CSF): G-CSF induced similar increase in chemiluminescence in control and patient PMN; GSF partially corrected the defective bactericidal activity; G-CSF did not affect the accelerated PMN apoptosis. These observations indicate that ALL children undergoing chemotherapy present PMN defective functions which are partially sensitive or even resistant to G-CSF. PMID- 9753059 TI - VLA-4 is involved in the engraftment of the human pre-B acute lymphoblastic leukaemia cell line NALM-6 in SCID mice. AB - Attachment of human pre-B leukaemic cells to human or murine bone marrow stromal cells in vitro is largely mediated by the beta1 integrin VLA-4 binding to VCAM-1. Cells subsequently migrate within the stroma, a process also involving VLA-4. A variant of the pre-B acute lymphoblastic leukaemia cell line NALM-6, designated 4A1, lacking expression of VLA-4, was generated by radiation-induced mutagenesis followed by several rounds of negative selection with immunomagnetic beads, fluorescence activated cell sorting and clonal expansion. In vitro assays using 4A1 cells showed reduced binding to, and migration under, the murine stromal line M2-10B4. Sublethally irradiated mice (n=19) with severe combined immunodeficiency were injected intravenously with NALM-6 cells. Animals developed signs of leukaemia with hind-limb paralysis at a median of 30 d (95% confidence interval 28-30). Although there were no gross abnormal findings at autopsy, histological analysis revealed extensive marrow replacement and focal liver infiltration with leukaemic blasts, which were confirmed to be of human origin by flow cytometry. 12 mice were injected with a similar number of cells from the VLA-4-negative variant cell line 4A1. Six mice developed signs of leukaemia after 43-74d, with the remaining six being free of signs of disease after > 100d (P<0.001). Mice in this group with leukaemia had a lower incidence of hind-limb paralysis and less leukaemic infiltration in the marrow, but in some cases had large tumour nodules elsewhere. After a single 500 microg intraperitoneal injection of anti-murine VCAM-1 monoclonal antibody (MK2.7), five additional mice were injected with an identical number of wild-type (VLA-4+) NALM-6. All animals developed signs of leukaemia after a similar period to those injected with wild-type NALM-6 only. These results demonstrate that the beta1 integrin VLA-4 is involved in the engraftment of the pre-B-cell leukaemic cell line NALM-6 in SCID mice, although the interaction with VCAM-1 is unlikely to be the sole explanation. PMID- 9753060 TI - Blastic variant of mantle cell lymphoma shows a heterogenous pattern of somatic mutations of the rearranged immunoglobulin heavy chain variable genes. AB - Mantle cell lymphoma is a distinct clinicopathological entity associated with t(11;14) and cyclin D1 overexpression. The majority of cases show uniform morphological and phenotypic features characterized by a monotonous proliferation of small-to-medium-sized irregular B cells that express CD5 and bright surface immunoglobulin IgM and IgD. By sequence analysis of the rearranged immunoglobulin heavy chain variable genes (VH), it has been shown that these lymphoma cells carry little if no somatic mutations, as described for the fetal CD5+ cells or B1 cells. Besides mantle cell lymphoma with classic histological features, a morphological variant of mantle cell lymphoma with blastic features and a more aggressive clinical course has been described. To investigate whether this variant is closely related, by the cell of origin, to typical cases, we analysed the presence and the pattern of somatic mutations of the VH genes in a series of nine cases diagnosed as such. Our cases of blastic mantle cell lymphomas rearrange most frequently VH4 and VH3 family genes. In three cases there was a complete homology to published germline genes, and a near complete homology was documented in another three. In contrast, the remaining three cases showed somatic mutations in their rearranged VH genes. Mutation analysis revealed evidence for antigen selection in one of these three cases. Taken together, these data are similar to those of normal adult-type B1 cells and those described for chronic lymphocytic leukaemia (CLL) but slightly different to those reported for classic mantle cell lymphoma. It is likely that blastic mantle cell lymphoma as well as CLL originates from adult-type B1 cells. More cases will need to be studied to determine whether classic mantle cell lymphoma is different from the blastic subtype and if it arises from fetal-type B1 cells. PMID- 9753061 TI - Integration of human herpesvirus 6 in a Burkitt's lymphoma cell line. AB - Human herpesvirus 6 (HHV-6) genome has been found in several human lymphoid malignancies, but configuration of the HHV-6 genome has not been well delineated. We established the HHV-6-positive, Epstein-Barr virus-negative Burkitt's lymphoma cell line Katata. In this study we investigated the status of the HHV-6 genome in Katata cells. Neither linear nor circular HHV-6 DNA was detected by Gardella gel analysis. The fluorescence in situ hybridization technique enabled us to directly visualize the integrated HHV-6 DNA at the single-cell level. Only one integrated site of viral DNA was detected in metaphase chromosomes and it was preferentially located at the long arm of chromosome 22 (22q13). Treatment of the cells with 12 O-tetradecanoyl-phorbol-13-acetate (TPA) or with calcium ionophore A23187 led to induction of the HHV-6 immediate-early gene as well as the late gene. Sodium n butyrate also gave rise to expression of the HHV-6 genes. The TPA inducibility was synergistically enhanced when combined with A23187 or n-butyrate. Our study provides, for the first time, an in vitro model system of latent HHV-6 infection whose genome is integrated into host DNA of lymphoma cells. PMID- 9753062 TI - Haematological response of patients with myelodysplastic syndrome to antithymocyte globulin is associated with a loss of lymphocyte-mediated inhibition of CFU-GM and alterations in T-cell receptor Vbeta profiles. AB - We have demonstrated that 44% of myelodysplastic syndrome (MDS) patients with cytopenia have a haematological response to antithymocyte globulin (ATG). Three ATG responders and two non-responders with refractory anaemia were further studied for lymphocyte-mediated inhibition of bone marrow using a standard CFU-GM assay. In responders, peripheral blood lymphocytes (PBL) added at a 5:1 ratio suppressed CFU-GM by 54+/-9% (P=0.04) and was reversed by ATG treatment. Pre treatment marrow depleted of CD3 lymphocytes, increased CFU-GM by 32% (P=0.02) in an ATG responder, but not in a non-responder. CD3 lymphocytes from 6-month post treatment marrow did not inhibit pre-treatment CFU-GM, indicating ATG had affected the T cells. Pre-treatment marrow depleted of CD8 lymphocytes, increased CFU-GM by 60% (P=0.01) and 49% (P=0.03) in two ATG responders, but not in a non responder. Inhibition required cell-cell interaction through MHCI. TCRVbeta families, analysed by SSCP, changed from clonal to polyclonal in one ATG responder after 6 months, but clones persisted in a non-responder. These results indicate patients with refractory anaemia who respond to ATG have CD8 T-cell clones that mediate MHCI-restricted suppression of CFU-GM which are replaced by polyclonal T cells that do not suppress CFU-GM after ATG treatment. PMID- 9753063 TI - Establishment and characterization of a mantle cell lymphoma cell line. AB - A mantle cell lymphoma (MCL) cell line (JeKo-1) was established from peripheral blood mononuclear cells of a patient with a large cell variant of MCL showing leukaemic conversion. JeKo-1 cells were Epstein-Barr virus negative and showed a B-cell phenotype with IgM+, IgD+, CD3-, CD5+, CD10-, CD19+, CD20+ and CD23-; they overexpressed cyclin D1, Bcl-2, c-Myc and Rb proteins. Bcl-1/J(H) gene rearrangement was confirmed by polymerase chain reaction, although karyotypic analysis showed 40/41 chromosomes devoid of apparent t(11;14)(q13;q32) translocation. JeKo-1 cells were highly tumourigenic in SCID mice. PMID- 9753064 TI - CD20-positive T-cell chronic lymphocytic leukaemia. AB - Although CD20 is considered to be a representative marker for B lymphocytes, the antigen is weakly expressed on a small subset of normal T lymphocytes. A 60-year old man developed pancytopenia and hepatosplenomegaly due to clonal proliferation of atypical lymphocytes that were weakly positive for CD20. The leukaemic cells were also positive for T-cell antigens such as CD2, CD3, CD5, CD7, CD8 and T-cell receptor (TCR) Vbeta8 and for activation antigens such as CD38 and HLA-DR, but were negative for CD19, CD21, CD22, CD25. Southern blot analysis revealed rearrangement of the TCR-beta gene and a germline configuration of the immunoglobulin heavy chain gene. This is the first report of a case of clonal expansion of CD20dim T lymphocytes. PMID- 9753065 TI - Association and clonal distribution of trisomy 12 and 13q14 deletions in chronic lymphocytic leukaemia. AB - The coexistence of trisomy 12 and deletions of chromosome 13 (13q12-q32) has rarely been observed in chronic lymphocytic leukaemia (CLL). Fluorescence in situ hybridization (FISH) performed on 600 consecutive CLL patients revealed the association of trisomy 12 and 13q14 deletion, of at least one of the three markers analysed (RB1, D13S319 and D13S25), in 55 cases (9% of 600 and 46% of 120 trisomy 12 cases). Trisomy 12 and isolated RB1 deletion were seen in 14/120 cases, trisomy 12 and D13S319/D13S25 deletion with diploid RB1 in 19/118, and trisomy 12 and deletion encompassing the three 13q markers studied in 22/118 cases. The heterogenous distribution of trisomy 12 and 13q deletions within the neoplastic B cells suggests that they are secondary rather than primary events in CLL leukaemogenesis. PMID- 9753066 TI - Coexpression of erythroid and megakaryocytic genes in acute erythroblastic (FAB M6) and megakaryoblastic (FAB M7) leukaemias. AB - There is evidence to suggest a close relationship between the erythroid and megakaryocytic lineages. Using RT-PCR, we evaluated the coexpression of erythroid and megakaryocytic genes in blasts from 25 acute myeloid leukaemia (AML) cases (FAB M1-M7) and three unclassifiable leukaemias with trilineage dysplasia (trilineal AML). All FAB M6 and M7 and trilineal leukaemias expressed mRNAs for alpha-globin, glycoprotein IIb (GpIIb), erythropoietin receptor (Epo-R) and thrombopoietin receptor (c-mpl), but not for myeloperoxidase (MPO) which in contrast was expressed in the other FAB-subtype leukaemias. These data support the hypothesis that blasts from M7 and M6 leukaemias may derive from (or represent) a common progenitor cell with resident bipotentiality towards the megakaryocytic and erythrocytic lineages. PMID- 9753067 TI - Dendritic cells derived from bone marrow and CD34+ selected blood progenitor cells of myeloma patients, cultured in serum-free media, do not contain the Kaposi sarcoma herpesvirus genome. AB - The aim of our study was to test if dendritic cells contain the KSHV genome. CD34+ peripheral blood progenitor cells (PBPC) and bone marrow mononuclear cells were cultured in X-VIVO 15 medium supplemented with GM-CSF and TNF-alpha in gas permeable containers. Dendritic cells were identified morphologically and immunophenotypically. The KSHV genome was not identified in any of the cases using a nested primer PCR approach. Serological analysis corroborated the molecular findings: no antibodies for KSHV were found in any of the multiple myeloma patients. These data are of importance when considering use of DC for therapeutic approaches in multiple myeloma. PMID- 9753068 TI - The activating splice mutation in intron 3 of the thrombopoietin gene is not found in patients with non-familial essential thrombocythaemia. AB - Essential thrombocythaemia (ET) is a condition of unknown aetiology characterized by sustained thrombocytosis in the absence of a detectable systemic cause. Although usually considered a clonal disease affecting myeloid cells, recent data indicate that a significant proportion of patients have polyclonal haemopoiesis. In some patients the thrombopoietin (TPO) levels are normal or raised. Recently a mutation has been described in the TPO gene in familial thrombocythaemia that results in elevated TPO levels. We have therefore screened 51 patients diagnosed with non-familial ET for the presence of this mutation, but it was not detected in any patient. The constitutional presence of this mutation is therefore unlikely to contribute to the pathogenesis of ET. PMID- 9753069 TI - The influence of HLA-matched sibling donor availability on treatment outcome for patients with AML: an analysis of the AML 8A study of the EORTC Leukaemia Cooperative Group and GIMEMA. European Organization for Research and Treatment of Cancer. Gruppo Italiano Malattie Ematologiche Maligne dell'Adulto. AB - To determine whether patients with a HLA-identical sibling donor have a better outcome than patients without a donor, an analysis on the basis of intention-to treat principles was performed within the framework of the EORTC-GIMEMA randomized phase III AML 8A trial. Patients in complete remission (CR) received one intensive consolidation course. Patients with a histocompatible sibling donor were then allocated allogeneic bone marrow transplantation (alloBMT), the patients without a donor were randomized between autologous BMT (ABMT) and a second intensive consolidation (IC2). 831 patients <46 years old and alive >8 weeks from diagnosis were included. HLA typing was performed in 672 patients. AlloBMT was performed during CR1 in 180 (61%) out of 295 patients with a donor. Another 38 patients were allografted: five in resistant disease, 14 during relapse and 19 in CR2. ABMT was performed in 130 (34%) out of 377 patients without a donor in CR1, in six (2%) patients during relapse and in 38 (10%) patients during CR2. The disease-free survival (DFS) from CR for patients with a donor was significantly longer than for patients without a donor (46% v 33% at 6 years; P=0.01, RR 0.78, 95% confidence interval 0.63-0.96). The overall survival from diagnosis for patients with a donor was longer, but not statistically significant, than for patients without a donor (48% v 40% at 6 years; logrank P=0.24). When patients were stratified according to prognostic risk groups, the same trend in favour of patients with a donor was seen for survival duration and the DFS remained significantly longer for this group of patients. PMID- 9753070 TI - Polymorphism in CD33 and CD34 genes: a source of minor histocompatibility antigens on haemopoietic progenitor cells? AB - Following bone marrow stem cell transplantation allo-responses against haemopoietic progenitor cells (HPC), causing graft rejection and graft-versus leukaemia effects, can be induced by donor T cells recognizing peptides derived from polymorphic endogenous proteins present in HPC. Since CD33 and CD34 are both expressed on HPC, we looked for genetic polymorphisms that might be the source of minor histocompatibility antigens (mHA) on such cells. Bone marrow from 14 donors and their HLA-identical recipients undergoing BMT for haematological malignancies were studied. Using non-radioactive single-strand conformation polymorphism analysis (cold SSCP) of complementary DNA encoding CD33 and CD34, three DNA polymorphisms, two in CD33 and one in CD34 were found and sequenced. Two were in non-coding regions, but in CD33, ATA or ATG at codon 183 resulted in an Ile or Met in the protein sequence. Nonapeptides derived from both alleles were predicted to bind to HLA A68.1. Thus two alleles of CD33 protein exist that could be mHA. With an alternate allele frequency of < 10%, allo-responses against CD33 would be uncommon after marrow transplantation. However, donors homozygous for this allele could be used to generate cytotoxic T cells against the frequent CD33 allele, for adoptive therapy of leukaemia. PMID- 9753071 TI - Autologous transplantation in chronic myeloid leukaemia using peripheral blood stem cells. AB - Forty-three patients with chronic myeloid leukaemia in first chronic phase were recruited to study intensive chemotherapy (idarubicin plus cytarabine; IdAC) followed by collection of peripheral blood stem cells (PBSC) in the recovery phase. PBSC autografting was performed on 32 patients. One patient died during mobilization and three died following autograft. All procedural deaths occurred in patients who received IdAc more than a year from diagnosis. Nine further patients died, eight following progression of CML. 72% of transplanted patients showed a major cytogenetic response but most cases have returned to Philadelphia positive haemopoiesis. 62% of autografted patients remain alive (median survival from diagnosis 52 months). Four of the 11 patients who did not receive a transplant remain in chronic phase. PMID- 9753072 TI - Molecular and biochemical characterization of JAK3 deficiency in a patient with severe combined immunodeficiency over 20 years after bone marrow transplantation: implications for treatment. AB - Severe combined immunodeficiency (SCID) comprises a heterogenous group of disorders that are fatal unless treated by bone marrow transplantation (BMT). The most common form of SCID (T-B+ SCID) is due to mutations of either the common gamma chain (gammac) or of gammac-coupled JAK3 kinase. We report an unusual JAK3 defect in a female who was successfully treated > 20 years ago with a BMT using her HLA-identical father as the donor. Persistence of genetically and biochemically defective autologous B cells, associated with reconstitution of cellular and humoral immunity, suggests that integrity of the gammac-JAK3 signalling pathway is not strictly required for immunoglobulin production. PMID- 9753073 TI - Telomerase activity and the expression of telomerase components in acute myelogenous leukaemia. AB - In 95 leukaemic cell samples from 66 patients with acute myelogenous leukaemia (AML) (47 de novo and 19 secondary AML) telomerase activity was determined and the expression of the telomerase components: telomerase reverse transcriptase (hTERT), telomerase RNA template (hTR) and telomerase-associated protein (TP1) evaluated by RT-PCR. Compared to peripheral blood mononuclear cells (PBMC) from normal adult 87% (82/95) of patient samples exhibited elevated telomerase activity hTERT, but not hTR and TP1 expression strongly correlated with the levels of telomerase activity (r=0.47, P<0.0001). The levels of telomerase activity were significantly higher at time of relapse or progression than at time of diagnosis (P=0.003), and correlated to CD34 expression and chromosomal abnormalities of leukaemic cells (P=0.01 and P=0.001 respectively). The rate and duration of complete remission (CR) did not correlate with the levels of telomerase activity at diagnosis. Among eight patients in first relapse, however, two of three with low levels of telomerase activity re-entered CR. whereas none of five patients with high telomerase activity achieved a second CR. Taken together, telomerase activation/up-regulation in AML is a disease progression associated event. Undifferentiated status and chromosomal aberration also lead to the up-regulation of telomerase activity in AML. PMID- 9753074 TI - PRAME, a gene encoding an antigen recognized on a human melanoma by cytolytic T cells, is expressed in acute leukaemia cells. AB - Gene PRAME was found to encode an antigen recognized on a human melanoma cell line by an autologous cytolytic T-lymphocyte clone. This gene is expressed at a high level in a very large fraction of tumours, such as melanomas, non-small-cell lung carcinomas, sarcomas, head and neck tumours and renal carcinomas. It is therefore a candidate for tumour immunotherapy even though some low expression is found in certain normal tissues. We tested by RT-PCR the expression of PRAME on more than 250 bone marrow or blood samples from patients with a haematological malignancy. Approximately 25% of the acute leukaemia samples were positive. Remarkably, all acute myeloblastic leukaemias that carried the chromosomal translocation t(8;21), which fuses the genes AML1 and ETO, expressed PRAME at a high level. PMID- 9753075 TI - Feto-maternal alloimmune thrombocytopenia: antenatal therapy with IVIgG and steroids. PMID- 9753076 TI - Factor V Leiden haplotypes in two homozygotes of Asian origin. PMID- 9753077 TI - Development of autoantibodies and factor VIII inhibitor in an HIV-infected haemophiliac following treatment with combination anti-retroviral therapy. PMID- 9753078 TI - Coincidence of neonatal alloimmune thrombocytopenia and maternal anti-D immunization: case report. PMID- 9753079 TI - Pulmonary embolism associated with varicella infection. PMID- 9753080 TI - Potentiation of dopamine-induced cAMP formation by group I metabotropic glutamate receptors via protein kinase C in cultured striatal neurons. AB - Metabotropic glutamate receptors have been shown to potentiate the cyclic adenosine monophosphate (cAMP) formation induced by activation of several receptors linked to adenylyl cyclase via Gs-protein. Here we show that, in primary cultures of striatal neurons, group I metabotropic receptors potentiate the cAMP formation induced by activation of D1-like dopamine receptors. Reverse transcription associated with polymerase chain reaction revealed that mGluR5, mGluR3, mGluR4 and mGluR7 are present in striatal cell cultures. The potentiation of cAMP formation is induced by the selective group I mGluRs agonist (S)-3,5 dihydroxyphenylglycine and by other non-selective mGluRs agonists with a typical group I-like pharmacology (quisqualate > ibotenate > 1-aminocyclopentane-1,3 dicarboxylic acid). The rank order potency of mGluRs agonists in potentiating cAMP formation correlates with their ability to induce inositol phosphates production; the potentiation of cAMP formation and the inositol phosphates production are blocked by the group I mGluRs antagonists (S)-4 carboxyphenylglycine and are not affected by group II antagonist 2S,3S,4S)-2 methyl-2-(carboxycyclopropyl)-glycine or group III antagonist (S)-2-amino-2 methyl-4-phosphonobutanoic acid. The potentiating mechanism involves the activation of protein kinase C, being mimicked by phorbol-12-myristate-13acetate and blocked by the specific protein kinase C inhibitors bisindolylmaleimide I and chelerythrine or by protein kinase C downregulation. Our results indicate that this interaction could have a functional importance in modulating the cAMP dependent transmission in the striatum. PMID- 9753081 TI - Effects of lesions of the nucleus basalis magnocellularis on the acquisition of cocaine self-administration in rats. AB - The nucleus basalis magnocellularis (NBM) is one element in the limbic cortical ventral striatal circuitry that has been implicated in reinforcement processes. The present study examined the involvement of the cholinergic neurons of the NBM in mediating aspects of cocaine reinforcement. Lesions of the NBM were made by injecting 0.01 M AMPA into the subpallidal basal forebrain. Following 4 days' recovery, rats were implanted chronically with catheters in the jugular vein. In three separate experiments, rats were trained to acquire cocaine self administration under a FR1 schedule of reinforcement at doses of 0.25, 0.083 and 0.028 mg/injection. A dose-effect function was also determined at the end of the acquisition experiments using five different doses of cocaine (0.009, 0.028, 0.083, 0.25, 0.50 mg/injection) and saline which were presented once daily in a Latin square design. There were no significant differences between groups in the acquisition of cocaine self-administration at any of the three doses studied (0.028, 0.083 and 0.25 mg/injection), although at the lowest dose, lesioned animals responded at greater levels on both active and inactive levers. However, a shift to the left in the cocaine dose-response function was observed revealing that the lesioned group self-administered significantly higher amounts of low doses of cocaine than control rats. These data suggest that the integrity of the NBM is not a critical determinant of the reinforcing effects of cocaine during the acquisition of self-administration of the drug, but that NBM-dependent cholinergic mechanisms may nevertheless interact with the neural substrates mediating the reinforcing properties of cocaine. The data are relevant to recent hypotheses of functional interactions between the dopaminergic system and the cholinergic NBM. PMID- 9753082 TI - Neuron-silicon junction with voltage-gated ionic currents. AB - We recorded the signals of firing Retzius neurons from Hirudo medicinalis by field-effect transistors. The axon stump of dissociated cells was attached to an open gate coated with concanavalin A. We observed a new type of neuron-transistor coupling: the extracellular voltage transients beneath the neuron were dominated by a negative peak during the rising phase of the action potential with a weaker positive transient in the falling phase. The biphasic response was opposite to the signal of capacitive coupling. We simulated the junction on the basis of the Hodgkin-Huxley equations. We found that the negative transient corresponded to an inward flow of sodium and the positive response to an outward flow of potassium. The field-effect transistors are able to probe the local flow of ionic currents in a membrane which is hidden in the region of cell adhesion. They may become a novel tool in neuroscience. PMID- 9753083 TI - Local cerebral glucose utilization in the AS/AGU rat: a mutant with movement disorders. AB - The AS/AGU mutant rat is characterized by a wide staggering gait and a movement disorder of the hindlimbs. Local cerebral glucose utilization in the brain was investigated using the [14C]2-deoxyglucose autoradiographic technique to map any functional alterations in the mutant AS/AGU (agu/agu) compared with Albino Swiss controls (+/+). Locomotor tests were also performed to confirm the phenotypic assignment of the animals. Statistically significant reductions in glucose utilization were apparent in 12 of the 44 regions examined in the AS/AGU animals. The regions showing the most significant differences (P < 0.01) from the control AS strain were the substantia nigra pars compacta (-23%) and medial geniculate body (-17%). Statistically significant decreases (P < 0.05 and P < 0.02) in glucose utilization ranging from -15 to -26% were also displayed in the superior colliculus superficial layer, auditory cortex, ventroposterior nucleus of the thalamus, molecular layer of the hippocampus, dentate gyrus, medial amygdaloid nucleus, median raphe nucleus, subthalamic nucleus, medial preoptic area of the hypothalamus and anterior hypothalamus. In no region studied was the mean value of glucose use in the AS/AGU rat greater than in the control animals. The results of this study complement previous behavioural and neurochemical characterization studies of this mutant, confirm that the disorder involves functional disturbances of the basal ganglia, and demonstrate the involvement of the limbic system and some sensory systems. PMID- 9753084 TI - Nitric oxide production is increased in the paraventricular nucleus of the hypothalamus of male rats during non-contact penile erections and copulation. AB - Male rats put in the presence of a receptive female rat that they can see, hear and smell, but cannot touch, show penile erection episodes. These non-contact erections occur concomitantly with an increase in nitric oxide production in the paraventricular nucleus of the hypothalamus, as detected by the increase in the NO2- and NO3- concentration in the paraventricular dialysate obtained from these males by in vivo microdialysis. NO2- concentration increased from 0.81+/-0.12 to 2.51+/-0.43 microM and that of NO3- from 4.50+/-0.73 to 8.31+/-2.3 microM. The NO2- increase was prevented by the nitric oxide synthase inhibitor NG-nitro-L arginine methylester (20 microg) given unilaterally in the paraventricular nucleus, which also prevented non-contact erections. In contrast, the nitric oxide scavenger haemoglobin (20 microg) prevented the NO2- increase, but not non contact erections; while the guanylate cyclase inhibitor methylene blue (20 microg) was ineffective on either response. NO2-and NO3- concentration was also increased in the paraventricular dialysate of male rats during in copula penile erections, that is, when sexual activity was allowed with the receptive females. As found with non-contact erections, NG-nitro-L-arginine methylester prevented NO2- increase and impaired copulatory behaviour; haemoglobin prevented NO2- increase only; and methylene blue was ineffective on either response. The present results confirm that nitric oxide is a physiological mediator of penile erection at the level of the paraventricular nucleus of the hypothalamus. PMID- 9753085 TI - Spatial organization of proprioception in the cat spinocerebellum. Purkinje cell responses to passive foot rotation. AB - This study examines the spinocerebellar locations of Purkinje cells that responded to passive foot rotations at the ankle joint in anaesthetized cats. Using a novel approach for mapping the locations of recorded cells from several animals onto an unfolded two-dimensional representation of the cortex, we found that cells distributed throughout the anterior-posterior extent of the spinocerebellar cortex, except in the most medial parts corresponding to zones a and b, were responsive to ankle joint rotation. The cell distributions revealed a clustering according to their response amplitudes, which showed evidence for both parasagittal and transverse banding. PMID- 9753086 TI - Interference with cellular energy metabolism reduces kynurenic acid formation in rat brain slices: reversal by lactate and pyruvate. AB - This study was designed to investigate the role of cellular energy metabolism in the de novo formation of the endogenous excitatory amino acid receptor antagonist, kynurenic acid. Using rat cortical tissue slices, the roles of glucose transport, glycolysis, tricarboxylic acid cycle intermediates and oxidative phosphorylation were studied. Inhibition of glucose utilization resulted in quantitatively similar decreases in kynurenine uptake, kynurenic acid production and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction, a marker of mitochondrial activity. The end product of glycolysis, pyruvate, as well as lactate, attenuated all three deficits. Pyruvate also significantly increased kynurenic acid formation in normal brain slices without affecting kynurenine uptake. Oxaloacetate and alpha-ketoglutarate (tricarboxylic acid cycle intermediates) were the only compounds tested which were capable of duplicating the effects of pyruvate, indicating that 2-oxoacids can stimulate kynurenic acid synthesis by acting as aminoacceptors in the enzymatic transamination of kynurenine. When the mitochondrial electron transport chain was blocked by specific inhibitors, coincubation with succinate restored the rate of MTT formazan formation to normal (except in the case of 3-nitropropionic acid), yet failed to prevent the resulting reduction in kynurenic acid synthesis. Conversely, pyruvate increased kynurenic acid production in the presence of all inhibitors (except cyanide), but did not attenuate the reduction in kynurenine uptake and MTT formazan formation. Taken together, these results demonstrate that interference with cellular energy metabolism causes mechanistically diverse, pronounced reductions in the cerebral neosynthesis of kynurenic acid, and that 2 oxoacids and lactate can effectively reverse most of these detrimental effects. PMID- 9753087 TI - Differential regulation of SHC proteins by nerve growth factor in sensory neurons and PC12 cells. AB - We have characterized some of the nerve growth factor (NGF) stimulated receptor tyrosine kinase (TrkA) signalling cascades in adult rat primary dorsal root ganglia (DRG) neuronal cultures and compared the pathways with those found in PC12 cells. TrkA receptors were phosphorylated on tyrosine residues in response to NGF in DRG neuronal cultures. We also saw phosphorylation of phospholipase Cgamma1 (PLCgamma1). We used recombinant glutathione-S-transferase (GST) PLCgamma1 SH2 domain fusion proteins to study the site of interaction of TrkA receptors with PLCgamma1. TrkA receptors derived from DRG neuronal cultures bound preferentially to the amino terminal Src homology-2 (SH2) domain of PLCgamma1, but there was enhanced binding with tandemly expressed amino- and carboxy terminal SH2 domains. The most significant difference in NGF signalling between PC12 cells and DRG was with the Shc family of adapter proteins. Both ShcA and ShcC were expressed in DRG neurons but only ShcA was detected in PC12 cells. Different isoforms of ShcA were phosphorylated in response to NGF in DRG and PC12 cells. NGF phosphorylated only one whereas epidermal growth factor phosphorylated both isoforms of ShcC in DRG cultures. Activation of the downstream mitogen activated protein (MAP) kinase, p42Erk2 was significantly greater than p44Erk1 in DRG whereas both isoforms were activated in PC12 cells. Blocking the MAP kinase cascade using a MEK1/2 inhibitor, PD98059, abrogated NGF dependent capsaicin sensitivity, a nociceptive property specific to sensory neurons. PMID- 9753088 TI - Association between the low threshold calcium spike and activation of NMDA receptors in guinea-pig substantia nigra pars compacta neurons. AB - The aim of this study was to examine the interaction between N-methyl-D-aspartate (NMDA) receptor activation and the low threshold calcium spike (LTS) of phasically firing neurons in the rostral part of the substantia nigra pars compacta (SNpc) in mid-brain slices. Bath perfusion of 10 microM NMDA gradually increased the LTS area and the effect reached a maximum after 6 min of perfusion. This enhancement of the LTS by NMDA was blocked both by a competitive and non competitive NMDA receptor antagonist, 50 microM D-AP5 and 10 microM MK801, respectively, demonstrating that this effect of NMDA was mediated through NMDA receptors. Prolonged exposure to increasing concentrations of NMDA (0.1-100 microM) progressively decreased the LTS area. The higher doses led to an irreversible marked depolarization and decrease of the membrane resistance. These results suggest that the LTS of SNpc neurons can trigger a NMDA receptor dependent response which may have physiological and pathological roles. PMID- 9753089 TI - Patterns of expression for the mRNA corresponding to the four isoforms of phospholipase Cbeta in mouse brain. AB - Ligand binding to neurotransmitter and hormone receptors which couple to the Gq subclass of GTP-binding protein leads to the activation of phospholipase Cbeta (PLCbeta) which hydrolyses phosphatidyl-inositol 4,5-bisphosphate, yielding a pair of second messengers, diacylglycerol and inositol 1,4,5-trisphosphate (IP3). The expression of PLCbeta1-4 mRNAs was comparatively examined by in situ hybridization in the mouse brain. In adults, PLCbeta1 mRNA was expressed predominantly in the telencephalon, including the cerebral cortex, hippocampus, amygdala, lateral septum and olfactory bulb, with little expression in most thalamic nuclei. PLCbeta2 mRNA was distributed in the white matter, suggesting its expression in non-neuronal cells, most likely oligodendrocytes. PLCbeta3 mRNA was specifically expressed in cerebellar Purkinje cells. The highest levels of PLCbeta4 mRNA were detected in Purkinje cells. High levels of PLCbeta4 mRNA were also found in the thalamus and medial septum, whereas weak signals were detected in most telencephalic regions, thus showing an expression pattern almost reciprocal to that of PLCbeta1 mRNA. During development, such characteristic regional expression of PLCbeta1 and PLCbeta4 were observed starting in late foetal stages, while specific expression of PLCbeta2 and PLCbeta3 appeared in early postnatal stages. We conclude that despite the existence of four PLCbeta isoforms, only one or two of them is expressed in individual neurons and glial cells. The distinct expression of PLCbetas provides a molecular basis for analysing the nature of the specific signal transduction pathway leading to the production of diacylglycerol and IP3 in distinct cell types and in different regions of the brain. PMID- 9753091 TI - Effects of transient oxygen-glucose deprivation on G-proteins and G-protein coupled receptors in rat CA3 pyramidal cells in vitro. AB - The role of guanosine triphosphate-binding proteins (G-proteins) in the generation of the outward current during transient oxygen-glucose deprivation (OGD) was investigated in CA3 pyramidal cells in rat hippocampal organotypic slice cultures using the single-electrode voltage-clamp technique with KMeSO4 filled microelectrodes. To simulate ischaemia, brief chemical OGD (2 mM 2 deoxyglucose and 3 mM NaN3 for 4-9 min) was used, which induced an outward K+ current associated with an increase in input conductance. OGD failed to induce the outward current under conditions where G-protein function was disrupted by loading cells with guanosine 5'-O-(2-thiodiphosphate) [GDPbetaS] or after prolonged injection of guanosine 5'-O(3-thiotdphosphate) [GTPgammaS]. However, in slices treated with pertussis toxin (PTX), OGD still elicited the outward current, indicating that PTX-insensitive G-proteins are involved. Consistent with this insensitivity to PTX, neither adenosine receptors nor GABA(B) (gamma aminobutyric acid) receptors, which operate via PTX-sensitive G-proteins, mediate the OGD-induced outward current. When adenosine receptors or GABA(B) receptors were blocked with 1,3-dipropyl-8-psulphophenylxanthine (DPSPX, 5 microM) or CGP 52 432 (10 microM), respectively, the OGD-induced response was not modified. The response also persisted following pretreatment of slice cultures with tetanus toxin to prevent vesicular release of neurotransmitters and neuromodulators from presynaptic terminals. Both PTX-sensitive and PTX-insensitive G-protein-mediated responses were suppressed during OGD. The inward current induced by the metabotropic glutamate receptor agonist 1 S, 3R-1-aminocyclopentane-1,3 dicarboxylate (1S,3R-ACPD) and the outward current elicited by adenosine or baclofen were strongly or completely attenuated. In contrast, the ionotropic alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) response was not affected. These findings suggest that during OGD there is a functional uncoupling of receptors from G-proteins, and a direct receptor-independent activation of PTX insensitive G-proteins leading to an increase in membrane K+ conductance. PMID- 9753090 TI - Amelioration of ischaemia-induced neuronal death in the rat striatum by NGF secreting neural stem cells. AB - The objective of the present study was to explore whether grafted immortalized neural stem cells, genetically modified to secrete nerve growth factor (NGF), can ameliorate neuronal death in the adult rat striatum following transient middle cerebral artery occlusion (MCAO). One week after cell implantation in the striatum, animals were subjected to 30 min of MCAO. Striatal damage was evaluated at the cellular level after 48 h of recirculation using immunocytochemical and stereological techniques. The ischaemic insult caused an extensive degeneration of projection neurons, immunoreactive for dopamine- and adenosine 3': 5' monophosphate-regulated phosphoprotein with a molecular weight of 32 kilodaltons (DARPP-32). 3H-Thymidine autoradiography demonstrated surviving grafted cells in the lesioned striatum in all transplanted rats. The loss of striatal projection neurons was significantly reduced (by an average of 45%) in animals with NGF secreting grafts, whereas control cells, not producing NGF, had no effect. The neuroprotective action of NGF-secreting grafts was also observed when the total number of striatal neurons immunopositive for the neuronal marker NeuN was quantified, as well as in cresyl violet-stained sections. The present findings indicate that administration of NGF by ex vivo gene transfer and grafting of neural stem cells can ameliorate death of striatal projection neurons caused by transient focal ischaemia. PMID- 9753092 TI - Neuroprotective role of ornithine decarboxylase activation in transient focal cerebral ischaemia: a study using ornithine decarboxylase-overexpressing transgenic rats. AB - Nuclear magnetic resonance imaging (MRI) was used to study dynamics of maturation and the size of ischaemic stroke lesions in rats with greatly increased activity of ornithine decarboxylase (ODC). Syngenic rats, either with or without chronic pre-ischaemic treatment with an ODC inhibitor, alpha-difluoromethylornithine (DFMO), as well as ODC-overexpressing transgenic rats were subjected either to transient middle cerebral artery (MCA) occlusion or permanent occlusion of the cortical branch of MCA. The two models were chosen to assess the role of ODC activity in damage caused by ischaemia and reperfusion, respectively. Diffusion of water was quantified by means of the trace of the diffusion tensor (D(av) = 1/3 Trace D) to assess the extent of energy failure and cytotoxic oedema, whereas the spin-spin relaxation time (T2) was used as a quantitative indicator of irreversible damage by MRI. Exposure to transient MCA occlusion resulted in significantly smaller stroke lesions in the ODC-overexpressing transgenic (246+/ 14 mm3) than in syngenic (320+/-9 mm3) or DFMO-treated (442+/-63 mm3) rats as determined 48 h after the occlusion. The differences in sizes were due to smaller lesions in the cortical tissue (transgenic vs. syngenic) or both in cortical and striatal regions (transgenic vs. DFMO-treated animals). The degree of irreversible oedema was greater in DFMO-treated rats than in syngenic or transgenic animals indicating accelerated development of a permanent damage in the absence of ODC induction. Cortical infarct following permanent MCA occlusion developed faster in the DFMO-treated than in syngenic or transgenic rats as the lesion sizes at 10 h were 26.2+/-4.3 mm3, 14.2+/-2.3 mm3 and 12.3+/-1.9 mm3, respectively. However, the stroke volumes by 48 h were not statistically different in the three animal groups. The present data demonstrate that ODC activation is an endogenous neuroprotective measure in transient cerebral ischaemia. PMID- 9753093 TI - Evidence for a contribution of lateral inhibition to orientation tuning and direction selectivity in cat visual cortex: reversible inactivation of functionally characterized sites combined with neuroanatomical tracing techniques. AB - We have previously reported that cells in cat areas 17 and 18 can show increases in response to non-optimal orientations or directions, commensurate with a loss of inhibition, during inactivation of laterally remote, visuotopically corresponding sites by iontophoresis of gamma-aminobutyric acid (GABA). We now present anatomical evidence for inhibitory projections from inactivation sites to recording sites where 'disinhibitory' effects were elicited. We made microinjections of [3H]-nipecotic acid, which selectively exploits the GABA re uptake mechanism, < 100 microm from recording sites where cells had shown either an increase in response to non-optimal orientations during inactivation of a cross-orientation site (n = 2) or an increase in response to the non-preferred direction during inactivation of an iso-orientation site with opposite direction preference (n = 5). Retrogradely labelled GABAergic neurons were detected autoradiographically and their distribution was reconstructed from series of horizontal sections. In every case, radiolabelled cells were found in the vicinity of the inactivation site (three to six within 150 microm). The injection and inactivation sites were located in layers II/III-IV and their horizontal separation ranged from 400 to 560 microm. In another experiment, iontophoresis of biocytin at an inactivation site in layer III labelled two large basket cells with terminals in close proximity to cross-orientation recording sites in layers II/III where disinhibitory effects on orientation tuning had been elicited. We argue that the inactivation of inhibitory projections from inactivation to recording sites made a major contribution to the observed effects by reducing the strength of inhibition during non-optimal stimulation in recurrently connected excitatory neurons presynaptic to a recorded cell. The results provide further evidence that cortical orientation tuning and direction selectivity are sharpened, respectively, by cross-orientation inhibition and iso-orientation inhibition between cells with opposite direction preferences. PMID- 9753094 TI - Fast confocal imaging of calcium released from stores in dendritic spines. AB - The emerging significance of calcium stores in neuronal plasticity and the assumed involvement of dendritic spines in long-term plastic properties of neurons have led us to examine the presence and possible regulation of calcium stores in dendritic spines. Immunohistochemical staining for ryanodine receptors was found in dendritic spines of cultured hippocampal neurons. Confocal microscopic imaging of calcium transients in dendritic spines of these neurons in response to caffeine allowed us to demonstrate an independent and unique calcium store in spines. The response to caffeine was blocked by thapsigargin and ryanodine, and maintained in calcium-free medium. The calcium stores were depleted faster in the spines than the dendrites. Furthermore, when calcium was released from stores under calcium-free conditions, and diffused passively between the spine and the dendrite, the length of the spine neck determined the degree of spine independence. Finally, the caffeine-sensitive ryanodine receptor linked calcium store was instrumental in regulating the response of neurons to glutamate. These results have important implications for understanding the roles of dendritic spines in neuronal integration and plasticity. PMID- 9753095 TI - Evidence that glypican is a receptor mediating beta-amyloid neurotoxicity in PC12 cells. AB - Docking of beta-amyloid fibrils to neuronal or glial cell membranes may be an early, necessary and intervenable step during the progression of Alzheimer's disease. Formation of neurofibrillary tangles and amyloid plaques as well as neurotoxicity and inflammation may be direct or indirect consequences. In an attempt to find a receptor that mediates those effects, we assessed rat pheochromocytoma PC12 cell 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) reduction after addition of beta-amyloid to the culture medium. Presence of competitive substances in the medium, cell-surface treatment and specific block of cellular synthesis pathways helped to identify the heparan sulphate moiety of a glycosylphosphatidylinositol-anchored protein likely to represent glypican as a possible receptor mediating beta-amyloid neurotoxicity. PMID- 9753096 TI - Mapping of neuronal networks underlying generalized seizures induced by increasing doses of pentylenetetrazol in the immature and adult rat: a c-Fos immunohistochemical study. AB - Previous studies from our group have shown that pentylenetetrazol (PTZ)-induced status epilepticus (SE) leads to age-dependent acute and long-term metabolic and circulatory changes in immature rats. In order to define the neural substrates involved in PTZ seizures according to age, the purpose of the present study was to map the areas of cellular activation during seizures of increasing severity in 10-day-old (P10), 21-day-old (P21) and adult rats. Seizures were induced by repetitive injections of subconvulsive doses of PTZ. The total dose received by the animals ranged from 4 to 125 mg/kg. These doses induced a variety of seizure profiles including absence-like, clonic seizures and SE. The cellular activation was measured as the density of c-Fos immunoreactive cells in animals at 2 h after the onset of the seizures. In P10 rats receiving a behaviourally non-active dose of PTZ, c-Fos immunoreactivity appeared only in the amygdala. The dose of 40 mg/kg that induced absence-like seizures led to a weak c-Fos expression in the medial thalamus, some cortical areas and globus pallidus. Clonic seizures reinforced labelling in the previous areas and induced a spread of c-Fos immunoreactivity to other cortical areas, thalamus, hypothalamus and some brainstem nuclei. At that age, only SE led to a widespread and stronger expression of c-Fos which was, however, totally lacking in the midbrain, and remained incomplete in the brainstem and forebrain limbic system, including the hippocampus. In P21 and adult rats, the inactive dose of PTZ induced c-Fos immunoreactivity in thalamus and hypothalamus. With absence-like seizures, c-Fos labelling spread to the cerebral cortex, amygdala, septum and some brainstem regions. With clonic seizures, immunoreactivity was reinforced in all areas already activated by absence-like seizures, and appeared in the striatum, accumbens, brainstem and hippocampus, except in CA1. After SE, c-Fos was strongly expressed in all brain areas. The intensity of c-Fos labelling was higher in most regions of P21 compared to adult rats. These data are in agreement with the immaturity of cellular and synaptic connectivity in P10 rats, the known greater sensitivity of rats to various kinds of seizures during the third week of life and the nature of the neural substrates involved in PTZ seizures. PMID- 9753097 TI - Experience-dependent depression of vibrissae responses in adolescent rat barrel cortex. AB - A short period of vibrissae deprivation in an adolescent (approximately 1 month old) rat can lead to depression of the cortical response to stimulation of the regrown vibrissae. In a barrel column representing the deprived vibrissa, depression is greater for neurons located close to the barrel column representing the spared vibrissa. One possible explanation is that the spared vibrissa produces heterosynaptic depression of the principal vibrissa response (Glazewski & Fox, 1996). To test this idea further, we compared the effect of depriving all vibrissae (no heterosynaptic influence at all) with depriving a single vibrissa (maximal heterosynaptic influence expected). In addition we tested the origin of the depression by recording from subcortical structures. After 7 days' deprivation and 6-8 days' regrowth, we tested the responses of barrel cortex cells, thalamic VPm neurons and trigeminal ganglion cells to stimulation of the regrown vibrissae. We found that depression was greater in cortex if a single vibrissa had been deprived than if all vibrissae had been deprived. (Average principal vibrissae responses in single vibrissae deprived animals were 36% of those in all vibrissae deprived animals for layer II/III and 41% for layer IV.) This implicates the spared vibrissae in actively down-regulating responses to the deprived vibrissae. However, some depression could also be produced in animals deprived of all vibrissae (layers II/III were 39% and layer IV 74% of control levels). These results indicate that simple withdrawal of activation has a depressive effect on responses but that depression is far greater if some active inputs remain. Neither form of deprivation had an effect on responses to principal vibrissa stimulation in the thalamus or trigeminal ganglion however, suggesting that depression originates in the cortex. Within the cortex, intracortical connections seem most affected as the greatest depression was found in layers II/III and in layer IV among cells responding at intermediate latencies (9-14 ms). PMID- 9753098 TI - Deficits and recovery of first- and second-order motion perception in patients with unilateral cortical lesions. AB - Unilateral lesions in the posterior parietal cortex can degrade motion perception in the contralesional visual hemifield. Our aim was to investigate whether deficits caused by cortical lesions may be different for first- and second-order motion perception, and to study the time scale of any potential recovery. In nine patients with circumscribed lesions mainly in the parietal and fronto-parietal cortex, thresholds for direction discrimination were measured for stimuli presented peripherally in their ipsi- and contralesional hemifield. Subjects had to identify the direction of a vertically moving object embedded in a background of dynamic random dot noise. The object consisted of various proportions of signal and noise dots. Signal dots were either (a) coherently moving in the same direction as the object (first-order), (b) stationary (second-order: drift balanced), or (c) coherently moving in the opposite direction (second-order: theta). Noise dots were flickering. Two patients showed significant threshold elevations for all three types of motion stimuli presented in their contralesional hemifield, while thresholds for ipsilesional targets were unaffected. Neither showed any selective deficit of first- versus second-order motion perception, but second-order motion was more impaired. Their lesions probably included the motion area V5-MT, which was spared in the other seven patients. One of the patients, who was retested several times during a 27-month postlesional period, showed complete recovery for first- and second-order motion direction discrimination, as well as for the detection of speed differences. PMID- 9753099 TI - Glutamate-dependent astrocyte modulation of synaptic transmission between cultured hippocampal neurons. AB - The idea that astrocytes merely provide structural and trophic support for neurons has been challenged by the demonstration that astrocytes can regulate neuronal calcium levels. However, the physiological consequences of astrocyte neuron signalling are unknown. Using mixed cultures of rat hippocampal astrocytes and neurons we have determined functional consequences of elevating astrocyte calcium levels on co-cultured neurons. Electrical or mechanical stimulation of astrocytes to increase their calcium level caused a glutamate-dependent slow inward current (SIC) in associated neurons. Microinjection of 1,2-bis(2 aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA) into astrocytes to prevent the stimulus-dependent increase in astrocyte calcium level, blocks the appearance of the neuronal SIC. Pharmacological manipulations indicate that this astrocyte dependent SIC is mediated by extracellular glutamate acting on N-methyl-D aspartate (NMDA) and non-NMDA glutamate receptors. Additionally, stimulation of astrocytes reduced the magnitude of action potential-evoked excitatory and inhibitory postsynaptic currents through the activation of metabotropic glutamate receptors. The demonstration that astrocytes modulate neuronal currents and synaptic transmission raises the possibility that astrocytes play a neuromodulatory role by controlling the extracellular level of glutamate. PMID- 9753100 TI - Regulatory roles of complexins in neurotransmitter release from mature presynaptic nerve terminals. AB - Complexins are presynaptic proteins whose functional roles in synaptic transmission are still unclear. In cultured rat hippocampal neurons, complexins are distributed throughout the cell bodies, dendrites and axons, whereas synaptotagmin I and synaptobrevin/VAMP-2, essential proteins for neurotransmitter release, accumulated in the synaptic-releasing sites as early as 1 week in culture. With a maturation of synapses in vitro, complexins also accumulated in the synaptic release sites and co-localized with synaptotagmin I and synaptobrevin/VAMP-2 after 3-4 weeks in culture. Complexins I and II were expressed in more than 90 and 70% of the cultured neurons, respectively; however, they were largely distributed in different populations of synaptic terminals. In the developing rat brain, complexins were distributed in neuronal cell bodies in the early stage of postnatal development, but gradually accumulated in the synapse-enriched regions with development. In mature presynaptic neurons of Aplysia buccal ganglia, injection of anticomplexin II antibody caused a stimulation of neurotransmitter release. Injection of recombinant complexin II and alphaSNAP caused depression and facilitation of neurotransmitter release from nerve terminals, respectively. The effect of complexin was reversed by a subsequent injection of recombinant alphaSNAP, and vice versa. These results suggest that complexins are not essential but have some regulatory roles in neurotransmitter release from presynaptic terminals of mature neurons. PMID- 9753101 TI - Glutamate receptor activation can trigger electrical activity in human glioma cells. AB - Cells from major types of gliomas, i.e. oligodendrogliomas and glioblastomas, are able to generate action potentials upon a current injection similar to neurons (Patt et al. (1996) Neuroscience, 71, 601-611; Labrakakis et al. (1997b) J. Neuropath. Exp. Neurol., 56, 243-254. Here, we report that activation of ionotropic glutamate receptors by the selective agonist, kainate, or by glutamate itself, depolarized the tumour cells in culture and living slices from tumour tissue, and can elicit volleys of action potentials, as recorded with the patch clamp technique. Sixty-six percent of the glioblastoma cells, 44% of the astocytoma and 86% of the oligodendroglioma cells responded to glutamate and the specific agonist of AMPA/kainate receptors, kainate. The involvement of non-NMDA (N-methyl-D-aspartate) receptors is further supported by the observation that both kainate and glutamate currents were blocked by CNQX (6-cyano-7 nitroquinoxaline-2,3-dione). The receptor activation was accompanied by an increase in cytosolic Ca2+, as recorded with a fura-2 microfluorometric system. The Ca2+ elevation was mediated by the activation of Ca2+ channels due to membrane depolarization. The presence of voltage-gated Ca2+ channels was confirmed by patch-clamp experiments. Taken together, these findings imply that the electrophysiological properties of glioma cells are more reminiscent of those of neurons than of glial cells. PMID- 9753102 TI - Serotonin depresses excitatory synaptic transmission and depolarization-evoked Ca2+ influx in rat basolateral amygdala via 5-HT1A receptors. AB - The actions of serotonin on rat basolateral amygdala neurons were studied with conventional intracellular recording techniques and fura-2 fluorimetric recordings. Bath application of 5-hydroxytryptamine (5-HT or serotonin) reversibly suppressed the excitatory postsynaptic potential in a concentration dependent manner without affecting the resting membrane potential and neuronal input resistance. Extracellular Ba2+ or pertussis toxin pretreatment did not affect the depressing effect of 5-HT suggesting that it is not mediated through activation of Gi/o protein-coupled K+ conductance. The sensitivity of postsynaptic neurons to glutamate receptor agonist was unaltered by the 5-HT pretreatment. In addition, the magnitude of paired-pulse facilitation was increased in the presence of 5-HT indicating a presynaptic mode of action. The effect of 5-HT was mimicked by the selective 5-HT1A agonist 8-hydroxy dipropylaminotetralin (8-OH-DPAT) and was blocked by the selective 5-HT1A antagonist 1-(2-methoxyphenyl)-4[4-(2-phthalimido)butyl]piperazine oxadiazol-3 yl]methyl]phenyl]-methanesulphonamide. In contrast, the selective 5-HT2 receptor antagonist ketanserin failed to affect the action of 5-HT. The effects of 5-HT and 8-OH-DPAT on the high K+-induced increase in [Ca2+]i were studied in acutely dissociated basolateral amygdala neurons. High K+-induced increase in [Ca2+]i was blocked by Ca2+-free solution and Cd2+ suggesting that Ca2+ entry responsible for the depolarization-evoked increase in [Ca2+]i occurred through voltage-dependent Ca2+ channels. Application of 5-HT and 8-OH-DPAT reduced the K+-induced Ca2+ influx in a concentration-dependent manner. The effect of 5-HT was completely abolished in slices pretreated with Rp-cyclic adenosine 3',5'-monophosphothioate (Rp-cAMP), a regulatory site antagonist of protein kinase A, suggesting that 5-HT may act through a cAMP-dependent mechanism. Taken together, these results suggest that functional 5-HT1A receptors are present in the excitatory terminals and mediate the 5-HT inhibition of synaptic transmission in the amygdala. PMID- 9753103 TI - The metabotropic glutamate receptor mGlu5 controls the onset of developmental apoptosis in cultured cerebellar neurons. AB - Cultured cerebellar granule cells grown in medium containing 10 mM K+ undergo apoptosis after 4-5 days in vitro (DIV), and, at that time, the activity of metabotropic glutamate (mGlu) receptors coupled to polyphosphoinositide (PI) hydrolysis begins to decline. In granule cells at 4 DIV, the mGlu receptor subtype mGlu5 was expressed at high levels. The expression of another PI-coupled mGlu receptor, the mGlu1a, was low at 4 DIV but increased during the following days. In cultures at 4-5 DIV, the few cells that already showed an apoptotic phenotype were devoid of mGlu5 receptors, but they all expressed mGlu1a receptors. The development of apoptosis was accelerated after treating the cultures with: (i) mGlu5 antisense oligonucleotides; (ii) the mixed mGlu receptor antagonist, (+)-alpha-methyl-4-carboxyphenylglycine; or (iii) the glutamate depleting enzyme, alanine aminotransferase. In contrast, an induced overexpression of mGlu5 receptors protected cultured granule cells against apoptotic death. We suggest that the activity of mGlu5 receptors supports cell survival, and a decline in the expression of mGlu5 receptors gives access to programmed cell death in cerebellar granule cells developing in primary cultures. PMID- 9753104 TI - Nerve growth factor and brain-derived neurotrophic factor increase neurotransmitter release in the rat visual cortex. AB - A number of experiments have shown that neurotrophins are involved in the development and plasticity of the visual cortex (Bonhoeffer, T., Curr. Op. Neurobiol., 6, 119, 1996). A possible mechanism underlying these effects is the neurotrophin modulation of synaptic transmission. We investigated whether nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) can modulate the release of neurotransmitter in the rat visual cortex at the peak of the critical period for plasticity (P23). The release of glutamate, acetylcholine and gamma aminobutyric acid (GABA) from visual cortical synaptosomes was analysed in continuous perfusion conditions. We found that NGF enhances the depolarization evoked release of glutamate (approximately 90%) and acetylcholine (approximately 35%) but not that of GABA. By contrast, BDNF enhances the depolarization-evoked release of all three neurotransmitters investigated (approximately 30%). BDNF and NGF were ineffective on basal release of neurotransmitters. The effect of NGF was not blocked by cholinergic antagonists atropine and mecamylamine. NGF and BDNF potentiation of transmitter release was strongly but not completely blocked by K252a, a tyrosine kinase inhibitor. The role of TrkA and p75NTR receptors was investigated in NGF-induced potentiation of glutamate release. Block of NGF binding to p75NTR using specific blocking antibodies (REX-IgG) slightly but significantly reduced the effect of NGF. Activation of TrkA in isolation by RTA IgG, an antibody that specifically activates TrkA, was less effective than activation of both receptors by NGF. These results show that neurotrophin action on neurotransmitter release was mostly mediated by Trk receptors with p75NTR having a little but significant positive role. Antigen blot analysis showed the presence of TrkA, TrkB and p75NTR receptors in the visual cortex. PMID- 9753105 TI - Survival and synaptogenesis of hippocampal neurons without NMDA receptor function in culture. AB - Physiological and morphological properties of cultured hippocampal neurons were measured to investigate whether NMDA receptors play a role in survival and differentiation. Neurons dissociated from mouse embryos with different NMDAR1 genotypes were grown in culture. Electrophysiological analysis verified the absence of NMDA receptor-mediated currents in neurons taken from homozygous mutant (NR1-/-) embryos. The number of surviving hippocampal neurons was 2.5-fold higher in cultures from the NR1-/- embryos compared with wild type (NR1 +/+) and heterozygous (NR1+/-) controls. Despite the lack of NMDA receptor function, NR1-/ neurons formed synapsin I-positive presynaptic boutons associated with MAP2ab positive dendrites in culture. Confocal microscopic analysis of Dil labelled neurons confirmed the presence of dendritic spines on NR1-/- neurons with 80% of the density found in NR1 +/+ neurons. These results suggest that the NMDA receptor has little effect on general features of neuronal differentiation. In contrast, there is clear effect on neuronal survival. This finding establishes neuron number in standard culture conditions as a measure of NMDA receptor activity. PMID- 9753106 TI - Parcellation of human mesial area 6: cytoarchitectonic evidence for three separate areas. AB - The mesial sector of primate area 6 is usually described as consisting of two distinct areas: the supplementary motor area (SMA or SMA proper) and the pre-SMA. Recent human brain imaging studies showed, however, that this subdivision is not completely satisfactory and that, most likely, SMA proper consists of two functionally distinct parts. In order to elucidate whether this hypothesis has an anatomical counterpart, we examined the cytoarchitectonic organization of human mesial area 6 in three brains of subjects deceased without any previous sign of neurological disorders. The data showed that human mesial area 6 consists of three separate cytoarchitectonic areas. Two of them are located mostly caudal to the vertical line transversing the anterior commissure (VCA line), the third one is located rostral to it. Given the location and some architectonic similarities between the two caudal areas, we named them caudal SMA (SMAc) and rostral SMA (SMAr). The area rostral to the VCA line is referred to as pre-SMA. The possible functional role of the three areas is discussed. PMID- 9753108 TI - Tyrosine kinases and synaptic transmission. PMID- 9753107 TI - Peripheral nerve insult induces NMDA receptor-mediated, delayed degeneration in spinal neurons. AB - Injury of a peripheral nerve gives rise to adaptive functional and structural alterations in spinal neurons. We report that the rearrangement of the spinal circuitry in response to sciatic nerve transection in adult rats involves a delayed mode of degeneration of lumbar spinal cord neurons. Nuclear fragmentation was detected by the TUNEL technique 7 days after sciatic neurectomy but not after 3 or 14 days. Dying cells were preferentially located in the ipsilateral superficial dorsal horn and expressed the neuronal cytoskeletal marker SMI-31. Degeneration was prevented by continuous systemic treatment with the NMDA receptor-antagonist MK-801. These data are supportive that apoptosis is induced in spinal neurons in a transsynaptic manner by an early signal from injured afferent fibres via activation of spinal NMDA receptors. PMID- 9753109 TI - Selective block of conduction in Y optic nerve fibres: significance for the concept of parallel processing. PMID- 9753110 TI - Adaptation in patterns of c-fos expression in the brain associated with exposure to either single or repeated social stress in male rats. AB - Intraspecific confrontation between male rats represents a biologically relevant form of social stress. C-fos expression has been used to map the pattern of neural activation following either a single (acute) or repeated (10 times) exposure of an intruder male to a larger male in the latter's home cage. These conditions induce high levels of aggressive interaction. Sixty minutes after a single defeat, there was intense c-fos expression (quantified using image analysis) in restricted areas of the basal forebrain (including lateral septum, bed nucleus of stria terminalis, lateral preoptic area, lateral hypothalamic area, paraventricular nucleus, and medial and central amygdala) as well as in the autonomic and monoaminergic nuclei of the brainstem (central grey, dorsal and median raphe, locus coeruleus and nucleus of the solitary tract). After the tenth defeat, this pattern was modified despite persistently high levels of aggression. Some areas in the forebrain (bed nucleus of stria terminalis, paraventricular nucleus and medial amygdala) continued to express increased c-fos; others (the septum, lateral hypothalamic area, lateral preoptic area and central amygdala) no longer expressed c-fos. The brainstem response was equally varied: the central grey and the raphe nuclei continued to respond after repeated defeat, whereas the solitary nucleus and locus coeruleus did not. On the other hand, there was no change in the behaviour of intruder rats after repeated defeat. This study shows the pattern of adaptation at a cellular level in the basal forebrain and brainstem to repeated defeat. As in our previous studies of repeated restraint, modulation in the expression of c-fos following repeated stress is highly regionally specific, suggesting that differential neural processing is involved in adaptation to social stress. PMID- 9753111 TI - Neural encoding of subject-object distance in a visual recognition system. AB - Domestic chicks follow a familiar (imprinted) object when it recedes. In behaving, imprinted chicks with no experience of objects at different distances, neuronal activity was recorded from the intermediate and medial part of the hyperstriatum ventrale (IMHV), a brain region crucial for the recognition memory underlying imprinting. We found that (i) some neurones responded equivalently, irrespective of the subject-object distance d (d-invariant); (ii) other neurones responded differently at different values of d (d-sensitive); (iii) these response types were found in monocular chicks and in chicks with both eyes exposed; (iv) the action potential shape of d-invariant neurones was different from that of other neurones and (v) the spontaneous firing rate of some neurones was correlated with locomotor activity. Taken together with previous findings, the results raise the possibility that IMHV has a major role to play in the sensory and motor-control aspects of imprinting in addition to its mnemonic functions. PMID- 9753112 TI - Increased lesion-induced sprouting of corticospinal fibres in the myelin-free rat spinal cord. AB - Myelin contains potent inhibitors of neurite growth which have been implicated in the failure of long-distance regeneration of nerve fibres within the CNS. These myelin-associated neurite growth inhibitors may also be involved in the stabilization of neural connections by suppressing sprouting and fibre growth. After lesions of the CNS in neonatal animals, extensive rearrangements of the remaining fibre systems have been observed. In the rat, this plasticity of neuronal connections is severely restricted following the first few weeks of postnatal life, coincident with the progression of myelination of the nervous system. A well-studied example of postnatal plasticity is the sprouting of one corticospinal tract (CST) into the denervated half of the spinal cord after unilateral motor cortex or pyramidal lesions. In the hamster and rat, significant CST sprouting is restricted to the first 10 postnatal days. Here we show that very extensive sprouting of corticospinal fibres occurs after deafferentations as late as P21 if myelination is prevented by neonatal X-irradiation in the rat lumbar spinal cord. Sprouted fibres from the intact CST cross the midline and develop large terminal arbors in the denervated spinal cord, suggesting the establishment of synaptic connections. Our results suggest that myelin and its associated neurite growth inhibitors play an important role in the termination of neurite growth permissive periods during postnatal CNS development. Corticospinal sprouting subsequent to lesions early in life, i.e. in the absence of myelin associated neurite growth inhibitors may explain the frequent occurrence of mirror movements in patients with hemiplegic cerebral palsy. PMID- 9753113 TI - Long-term protection of the rat nigrostriatal dopaminergic system by glial cell line-derived neurotrophic factor against 6-hydroxydopamine in vivo. AB - Glial cell-line-derived neurotrophic factor (GDNF) has been shown to enhance the survival of dopaminergic neurones both in vitro and in vivo, and to protect the rodent dopaminergic system from neurotoxic damage. However, most previous studies have only examined the short-term protective effects of GDNF. We have investigated the long-term effects of GDNF on a 6-hydroxydopamine (6-OHDA) induced lesion of the rat medial forebrain bundle (MFB), which results in complete and irreversible destruction of the nigrostriatal pathway, and is a robust model of Parkinson's disease. GDNF was administered ipsilaterally above the substantia nigra and into the lateral ventricle immediately before a unilateral 6-OHDA injection into the MFB. The effects of GDNF were examined in vivo by behavioural testing and positron emission tomography (PET) at weekly intervals, for 12 weeks. GDNF prevented the development of amphetamine-induced rotations at all time-points. PET studies, using [11C]-RTI-121 as a tracer for the dopamine transporter, indicated that GDNF prevented 6-OHDA-induced reduction of dopamine reuptake sites in the ipsilateral striatum. Post-mortem neurochemical analysis at 13 weeks after surgery found that GDNF significantly inhibited 6-OHDA induced loss of dopamine, 3,4-dihydroxyphenylacetic acid and homovanillic acid in the ipsilateral striatum. Immunocytochemistry showed that GDNF reduced 6-OHDA induced loss of tyrosine hydroxylase-positive neurones in both the substantia nigra pars compacta and ventral tegmental area. We have shown that a single treatment with GDNF can confer long-term protective effects against a 6-OHDA lesion, which suggests that this factor may be useful for the treatment of Parkinson's disease. PMID- 9753114 TI - Functional diversity of synaptic AMPA/KA receptors from rat as revealed by subtype-specific antagonists. AB - Subtype-specific pharmacological compounds represent important tools to identify the molecular components of synaptically activated glutamate receptors in central neurones. Here, we utilized a collection of subtype-specific antagonists and modulators to investigate the functional profile of glutamate receptors in identified synapses in thin slices of the cerebellum, hippocampus and brain stem. During whole-cell patch-clamp recordings alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionate/kainate (AMPA/KA) receptor-mediated synaptic currents (EPSCs) in cerebellar Purkinje cells were (i) prolonged by 100 microM cyclothiazide, (ii) not significantly changed after preincubation in 10 microM concanavalin A, (iii) not affected by 1 microM Evans Blue or polyamine toxin analogue N-(4 hydroxyphenylpropanolyl)-spermine (NHPPS), but (iv) significantly reduced by high (> or = 100 microM) concentrations of Evans Blue. These pharmacological properties were distinct from those observed in hippocampal granule cells and brain stem interneurones and markedly different from those of recombinant glutamate receptor channels GluR1-GluR6 previously investigated in heterologous expression systems. PMID- 9753115 TI - Incorporation of mouse neural progenitors transplanted into the rat embryonic forebrain is developmentally regulated and dependent on regional and adhesive properties. AB - During development, telencephalic neural progenitors acquire positional specification and give rise to distinct structures such as the striatum and cortex. Here, we examine, in vivo, the influence of developmental stage, cell surface molecules and regional differences along the dorso-ventral and antero posterior axes on the selective incorporation of neural progenitors derived from different regions of the developing brain, utilizing a cross-species in utero transplantation paradigm. Striatal progenitors derived from the embryonic day (E) 12-14 mouse lateral ganglionic eminence (LGE) were observed consistently to incorporate into the developing striatum as early as 24-48 h following intraventricular injection into the E15-17 rat host. By removing cell-surface molecules from the LGE progenitors, the pattern of incorporation was remarkably different with no preferential striatal incorporation. Cortical progenitors with intact cell-surface molecules, by contrast, displayed little telencephalic (including striatal) incorporation as compared with precursors from the LGE. However, both progenitors from cortex and LGE incorporated widely into diencephalic and mesencephalic structures. The capacity for integration of precursors derived from the LGE and cortex gradually decreased during development of the host and was minimal in the postnatal day (P) 1 host. Unlike the telencephalic precursors, the vast majority of progenitors derived from the midbrain and cerebellar primordium (with cell-surface molecules intact) incorporated into diencephalic and midbrain nuclei with only a few cells observed in the telencephalon. These results demonstrate that incorporation of neural progenitors across the ventricular wall in the embryonic host is strictly developmentally regulated, dependent on their position along the antero-posterior axes and in the case of progenitors from the LGE is mediated by cell-surface molecules expressed on the transplanted cells. PMID- 9753116 TI - Role of the cerebellum in reaching movements in humans. I. Distributed inverse dynamics control. AB - This study focuses on the role of the motor cortex, the spinal cord and the cerebellum in the dynamics stage of the control of arm movement. Currently, two classes of models have been proposed for the neural control of movements, namely the virtual trajectory control hypothesis and the acquisition of internal models of the motor apparatus hypothesis. In the present study, we expand the virtual trajectory model to whole arm reaching movements. This expanded model accurately reproduced slow movements, but faster reaching movements deviated significantly from the planned trajectories, indicating that for fast movements, this model was not sufficient. These results led us to propose a new distributed functional model consistent with behavioural, anatomical and neurophysiological data, which takes into account arm muscles, spinal cord, motor cortex and cerebellum and is consistent with the view that the central nervous system acquires a distributed inverse dynamics model of the arm. Previous studies indicated that the cerebellum compensates for the interaction forces that arise during reaching movements. We show here how the cerebellum may increase the accuracy of reaching movements by compensating for the interaction torques by learning a portion of an inverse dynamics model that refines a basic inverse model in the motor cortex and spinal cord. PMID- 9753117 TI - Role of the cerebellum in reaching movements in humans. II. A neural model of the intermediate cerebellum. AB - The cerebellum is essential for the control of multijoint movements; when the cerebellum is lesioned, the performance error is more than the summed errors produced by single joints. In the companion paper (Schweighofer et al., 1998), a functional anatomical model for visually guided arm movement was proposed. The model comprised a basic feedforward/feedback controller with realistic transmission delays and was connected to a two-link, six-muscle, planar arm. In the present study, we examined the role of the cerebellum in reaching movements by embedding a novel, detailed cerebellar neural network in this functional control model. We could derive realistic cerebellar inputs and the role of the cerebellum in learning to control the arm was assessed. This cerebellar network learned the part of the inverse dynamics of the arm not provided by the basic feedforward/feedback controller. Despite realistically low inferior olive firing rates and noisy mossy fibre inputs, the model could reduce the error between intended and planned movements. The responses of the different cell groups were comparable to those of biological cell groups. In particular, the modelled Purkinje cells exhibited directional tuning after learning and the parallel fibres, due to their length, provide Purkinje cells with the input required for this coordination task. The inferior olive responses contained two different components; the earlier response, locked to movement onset, was always present and the later response disappeared after learning. These results support the theory that the cerebellum is involved in motor learning. PMID- 9753118 TI - A presynaptic N-methyl-D-aspartate autoreceptor in rat hippocampus modulating amino acid release from a cytoplasmic pool. AB - A possible role of the N-methyl-D-aspartate receptor (NMDA-R) as a presynaptic autoreceptor was investigated using Percoll-purified hippocampus nerve terminals (synaptosomes). This preparation contained only a neglectable amount of postsynaptic structures. Two main effects of NMDA were observed. First, NMDA dose dependently (10-100 microM) and in the absence of Mg2+, stimulated basal release of aspartate and glutamate, but not of GABA. MK801 (10 microM), an open NMDA-R channel blocker, reduced this effect even below control levels, indicating endogenous NMDA-R activation. By superfusing synaptosomes, which prevents a tonic receptor occupation, also basal GABA release was stimulated by NMDA. The NMDA induced potentiation of amino acid superfusate levels was blocked both by MK801 and Mg2+ (1 mM), was slow in onset and returned to baseline after NMDA-removal. The NMDA-effect was also found in the absence of extracellular Ca2+, suggesting that amino acids were released from a non-vesicular (cytoplasmic) pool. Secondly, in KCl-depolarized synaptosomes exposed to 1 mM Mg2+, NMDA did not affect the release of the amino acids. MK801, however, reduced the KCl-evoked Ca2+ independent release of aspartate and glutamate, but not of GABA. L-trans-PDC, the selective inhibitor of the glutamate/aspartate transporter, prevented this MK801 effect, suggesting a coupling between NMDA-Rs and these transporters. These data provide evidence for a presynaptic NMDA autoreceptor in rat hippocampus. We speculate on the role of this NMDA-R to depolarize the presynaptic membrane by Na+-entry, which may induce reversal of amino acid transporters and thereby releasing amino acids from a cytoplasmic pool. PMID- 9753119 TI - Synaptic clustering of GABA(C) receptor rho-subunits in the rat retina. AB - Polyclonal antibodies which recognize the rho-subunits of the GABA(C) receptor were applied to sections of the rat retina. Strong punctate immunoreactivity was found in the inner plexiform layer (IPL), which was shown by electron microscopy to represent a clustering of the GABA(C) receptors at synaptic sites. During postnatal development diffuse rho-immunoreactivity was first observed at postnatal day P3. Distinct labelling of bipolar cells appeared at P7 and punctate, synaptic labelling was observed at P10. In order to show that the rho immunoreactive puncta coincide with the axons of bipolar cells, double immunostainings of retinal sections with an antiserum against syntaxin 3 and with the rho-antiserum were performed. The experiments showed that rho-immunoreactive puncta are preferentially located on the axon terminals of rod and cone bipolar cells. In order to determine whether GABA(C) receptor rho-subunits coassemble with GABA(A) receptor subunits, double-labelling experiments were performed with subunit specific antisera. Punctate, putative synaptic clustering was observed with all antisera applied, however, GABA(C) receptor expressing puncta did not coincide with GABA(A) receptor containing puncta. This suggests that there are no synaptic GABA receptors in the retina in which GABA(A) and GABA(C) receptor subunits are coassembled. Similar double-labelling experiments were also performed to find out whether GABA(C) receptors and glycine receptors are colocalized. They were clustered at different synapses. This suggests that synaptic GABA(C) receptors consist of rho-subunits and are not coassembled with GABA(A)- or glycine-receptor subunits. PMID- 9753120 TI - Characterization of the multisynaptic neuronal control of the rat pineal gland using viral transneuronal tracing. AB - Knowledge of the polysynaptic pathway conveying photic information to the pineal gland is based upon studies employing lesions, knife cuts and classical tracers. In the present investigation we used viral transneuronal tracing to re-examine the organization of this circuitry. This was accomplished by injecting a neurotropic alpha herpesvirus (pseudorabies virus) into the gland and localizing viral antigen in infected neurones at various postinoculation intervals. This approach is based upon the demonstrated ability of this virus to invade axon terminals, replicate in neurones and pass retrogradely through a multisynaptic circuit. Immunohistochemical localization of viral antigen revealed the progressive appearance of infected neurones in the superior cervical ganglion (SCG), intermediolateral nucleus of the upper thoracic spinal cord (IML), parvicellular subdivisions of the hypothalamic paraventricular nucleus (PVN), and the suprachiasmatic nucleus (SCN). Other infected cell groups known to project to the IML also became infected. Infection of the PVN reproducibly involved neurones in the dorsal, medial and lateral parvicellular subdivisions and preceded the appearance of infected neurones in the SCN and other regions of hypothalamus. Topographic analysis of virus infected neurones within the SCN revealed differential infection of SCN subdivisions that suggested topography in the projection of the SCN to the PVN. Removal of the SCG eliminated infection within the aforementioned circuitry and revealed a parasympathetic innervation from the sphenopalatine ganglion. The data provide further detail on the cellular identity and synaptology of neural circuitry controlling the rhythmic secretion of melatonin by the rat pineal gland. PMID- 9753121 TI - Corneal innervation and sensitivity to noxious stimuli in trkA knockout mice. AB - Most primary sensory neurones depend on neurotrophins for survival. Mutant mice in which TrkA, the high-affinity receptor for nerve growth factor (NGF), has been inactivated lack nociceptive neurones in sensory ganglia and do not respond to noxious stimuli. The cornea of the eye is innervated by trigeminal neurones that are activated by noxious mechanical, thermal and chemical stimuli. In the human cornea, these stimuli evoke only sensations of pain. We have analysed the innervation pattern and the response to noxious stimulation of the cornea of trkA (-/-) mutant mice. Corneal nerves were stained with the gold chloride impregnation method. Corneal sensitivity to noxious stimuli was assessed by counting blinking movements evoked by von Frey hairs, topical application of saline at different temperatures and application of acetic acid and capsaicin at different concentrations. In the cornea of trkA (-/-) mutant animals, we observed a drastic reduction in the number of nerve trunks and branches in the corneal stroma. Furthermore, quantitative analysis of the number of thin nerve terminals revealed a marked decrease in the corneal epithelium of trkA (-/-) mice when compared to those present in wild type and trkA (+/-) animals. The blinking response of trkA (-/-) mice to mechanical, thermal and chemical noxious stimuli was also significantly reduced. These results indicate that the population of corneal sensory neurones is markedly depleted in trkA (-/-) mutant mice. However, a small portion of corneal sensory neurones survive in these mice suggesting that they may be NGF independent. On the basis of our results, we propose that these surviving cells are polymodal nociceptive neurones, sensitive to mechanical stimulation, noxious heat and acid. PMID- 9753122 TI - Eye position encoding in the macaque posterior parietal cortex. AB - In two previous studies, we had demonstrated the influence of eye position on neuronal discharges in the middle temporal area, medial superior temporal area, lateral intraparietal area and area 7A of the awake monkey (Bremmer et al., 1997a,b). Eye position effects also have been found in visual cortical areas V3A and V6 and even in the premotor cortex and the supplementary eye field. These effects are generally discussed in light of a coordinate transformation of visual signals into a non-retinocentric frame of reference. Neural network studies dealing with the eye position effect succeeded in constructing such non retinocentric representations by using model neurones whose response characteristics resembled those of 'real' neurones. However, to our knowledge, response properties of real neurones never acted as input into these neural networks. In the present study, we thus investigated whether, theoretically, eye position could be estimated from the population discharge of the (previously) recorded neurones and, if so, we intended to develop an encoding algorithm for the position of the eyes in the orbit. The optimal linear estimator proved the capability of the ensemble activity for determining correctly eye position. We then developed the so-called subpopulation encoding of eye position. This algorithm is based on the partition of the ensemble of neurones into two pairs of subpopulations. Eye position is represented by the differences of activity levels within each pair of subpopulations. Considering this result, encoding of the location of an object relative to the head could easily be accomplished by combining eye position information with the intrinsic knowledge about the retinal location of a visual stimulus. Taken together, these results show that throughout the monkey's visual cortical system information is available which can be used in a fairly simple manner in order to generate a non-retinocentric representation of visual information. PMID- 9753123 TI - MAP2d mRNA is expressed in identified neuronal populations in the developing and adult rat brain and its subcellular distribution differs from that of MAP2b in hippocampal neurones. AB - The brain microtubule-associated protein MAP2 family is composed of high molecular-weight (MAP2a and MAP2b) and low-molecular-weight (MAP2c and MAP2d) isoforms. The common C-terminal region of HMW MAP2 and MAP2c contains three repeated microtubule-binding domains while MAP2d comprises four repeats. MAP2c mRNA is known to be expressed at high levels in the immature brain. We show that in the brains of rat pups, MAP2c mRNAs are indeed expressed at high levels compared with MAP2d. However, in adult rat brains, MAP2d mRNA levels are higher than MAP2c. In order to identify the neural cells expressing MAP2d, we used in situ hybridization. In vivo, we show that MAP2d mRNA is expressed in well identified neuronal populations of the brain. In primary cultures of hippocampal neurones, double-labelling experiments confirm that MAP2d is clearly expressed in neurones. We also evaluated in this study the subcellular distribution of the MAP2d mRNAs in cultured hippocampal neurones and we report that in contrast with MAP2b mRNAs, mostly localized in dendrites, MAP2d mRNAs are essentially located in neuronal cell bodies. PMID- 9753124 TI - The neuronal alpha6 subunit forms functional heteromeric acetylcholine receptors in human transfected cells. AB - We examine some of the biological and physiological properties of the avian alpha6 neuronal nicotinic acetylcholine receptor (nAChR) subunit. We show here that, beginning at embryonic day 5, alpha6 mRNA is abundantly expressed in the developing chick neuroretina, where it coexists with other nicotinic receptor subunit mRNAs such as alpha3, beta2 and beta4. In contrast, alpha6 mRNA is absent from the optic tectum and from the peripheral ganglia. Despite numerous efforts, the alpha6 subunit has long failed the critical test of functional reconstitution. Here we use patch-clamp techniques and confocal laser microscopy to measure ACh-activated currents and nicotine-elicited Ca2+ transients in human BOSC 23 cells transfected with chick alpha6 in combination with other chick nAChR neuronal subunits. Heterologously expressed alpha6 and beta4 subunits form functional heteromeric nAChRs, which are permeable to Ca2+ ions and blocked by the nicotinic antagonist methyllycaconitine (10 microM). Likewise, ACh elicits measurable currents in cells transfected with alpha6 and beta2. Hill analysis of the dose-response curves in cells transfected with alpha3, beta4 and alpha6 cDNAs, suggests the assembly of functional alpha3beta4alpha6 receptor, with an apparent affinity for ACh threefold lower than alpha3beta4. Our results indicate that alpha6-containing nAChRs assemble in heterologous expression systems and are probably present in retinal cells. PMID- 9753125 TI - Block by picrotoxin of a GABAergic chloride channel expressed on crayfish muscle after axotomy. AB - Outside-out patches containing a gamma-aminobutyric acid (GABA)-activated chloride channel expressed after axotomy on crayfish deep extensor abdominal muscle were excised. GABA and the blocker picrotoxin (PTX) were applied to the patches using a liquid filament switch to study the effects of picrotoxin on the GABA-elicited currents. Coapplication of GABA and PTX resulted in a reduction of the current amplitude compared with that elicited by the same GABA concentration alone. This reduction of the amplitude was dependent on both the GABA and PTX concentrations. The rise time of the current decreased after coapplication of GABA and PTX. Evaluation of the single channel currents and off-currents in the presence of GABA and PTX showed a dramatic shortening of the burst duration of the channel. The open time distributions were not altered, whereas in the closed time distributions a new closed time was apparent in presence of PTX. Preincubation with PTX prior to the GABA pulse resulted in an increase of the rise time. This effect was dependent on the PTX concentration only. Possible mechanisms are discussed to explain the effects of PTX and are implemented into the existing molecular reaction scheme of the channel. PMID- 9753126 TI - Effect of heparin-binding growth-associated molecule (HB-GAM) on synaptic transmission and early LTP in rat hippocampal slices. AB - Heparin-binding growth-associated molecule (HB-GAM) is an 18-kDa developmentally regulated protein, which promotes neurite outgrowth, axonal guidance and synaptogenesis through interaction with cell-surface heparan-sulphate proteoglycans. We have studied the effect of HB-GAM on synaptic transmission and long-term potentiation (LTP) in the area CA1 of rat hippocampal slices, where HB GAM mRNA is expressed in an activity-dependent manner. Injection of recombinant HB-GAM into the dendritic area inhibited tetanus-induced LTP without affecting baseline synaptic responses or the N-methyl-D-aspartate (NMDA)-receptor mediated transmission. HB-GAM did not depotentiate tetanus-induced LTP or prevent heterosynaptic LTP induced by application of tetraethylammonium (TEA), indicating that the effect was limited to early, synapse-specific stages of LTP induction. These results suggest that HB-GAM is involved in the regulation of synaptic plasticity in hippocampus. PMID- 9753127 TI - Visuo-motor transformations for arm reaching. AB - Visuomanual co-ordination requires the merging of ocular and arm information in a common frame of reference. Here we consider behavioural evidence in humans for the use of a viewer-centred frame in the specification of end point positions of reaching. We then review anatomical and neurophysiological data in the non-human primate that indicate a prominent role of the parietal cortex in the process of multisensory fusion that leads to egocentric representations of space. Finally, we discuss the functional anatomy of the human parietal cortex in visuomanual co ordination as revealed by neuroimaging. PMID- 9753128 TI - Somatostatin receptor subtypes sst1 and sst2 elicit opposite effects on the response to glutamate of mouse hypothalamic neurones: an electrophysiological and single cell RT-PCR study. AB - We have previously shown that somatostatin can either enhance or decrease AMPA/kainate receptor-mediated responses to glutamate in mouse-dissociated hypothalamic neurones grown in vitro. To investigate whether this effect is due to differential activation of somatostatin (SRIF) receptor subtypes, we compared modulation of the response to glutamate by SRIF with that induced by CH-275 and octreotide, two selective agonists of sst1 and sst2/sst5 receptors, respectively. Somatostatin either significantly decreased (49%) or increased (30%) peak currents induced by glutamate, and was ineffective in the remaining cells. Only the decreased response was obtained with octreotide, whereas only increased responses were elicited by CH-275 (47 and 35% of the tested cells, respectively). Mean amplitude variations under somatostatin or octreotide on the one hand, and under somatostatin or CH-275 on the other hand, were equivalent. Pertussis toxin pretreatment significantly decreased the number of cells inhibited by somatostatin or octreotide, but had no effect on the frequency of neurones showing increased sensitivity to glutamate during somatostatin or CH-275 application. About half of the neurones tested by single cell reverse transcriptase polymerase chain reaction (RT-PCR) expressed only one sst receptor (sst1 in 26% and sst2 in 22% of studied cells). Out of the remaining neurones, 34% displayed neither sst1 nor sst2 mRNAs, whereas 18% showed a simultaneous expression of both mRNA subtypes. Expression of sst1 or sst2 mRNA subtypes matched totally with the effects of somatostatin on sensitivity to glutamate in 79% of the neurones processed for PCR after recordings. These data show that pertussis toxin-insensitive activation of the sst1 receptor subtype mediates somatostatin-induced increase in sensitivity to glutamate, whereas decrease in the response to glutamate is linked to pertussis toxin-sensitive activation of the sst2 receptor subtype. PMID- 9753129 TI - Suppression of kindling epileptogenesis in rats by intrahippocampal cholinergic grafts. AB - Selective immunolesioning of the basal forebrain cholinergic system by 192 IgG saporin, which leads to a dramatic loss of the cholinergic innervation in cortical and hippocampal regions, facilitates the development of hippocampal kindling in rats. The aim of the present study was to explore whether grafted cholinergic neurones are able to reverse the lesion-induced increase of seizure susceptibility. Intraventricular 192 IgG-saporin was administered to rats which 3 weeks later were implanted with rat embryonic, acetylcholine-rich septal-diagonal band tissue ('cholinergic grafts') or cortical tissue/vehicle ('sham grafts') bilaterally into the hippocampal formation. After 3 months, the grafted animals as well as non-lesioned control rats were subjected to daily hippocampal kindling stimulations. In the animals with cholinergic grafts, which had reinnervated the hippocampus and dentate gyrus bilaterally, there was a marked suppression of the development of seizures as compared with the hyperexcitable, sham-grafted rats. This effect was significantly correlated to the density of the graft-derived cholinergic innervation of the host hippocampal formation. The kindling rate in the rats with cholinergic grafts was similar to that in non-lesioned controls. These results provide further evidence that the intrinsic basal forebrain cholinergic system dampens kindling epileptogenesis and demonstrate that this function can be exerted also by grafted cholinergic neurones. PMID- 9753130 TI - Stimulation of 5-HT1A receptors in the dorsal raphe reverses the impairment of spatial learning caused by intrahippocampal scopolamine in rats. AB - This study investigated the effect of stimulating 5-HT1A receptors in the dorsal raphe on the impairment of learning caused by 4 microg/microL scopolamine injected in the CA1 region of the dorsal hippocampus in rats performing a two platform spatial discrimination task. At 1 (but not 0.2) microg/0.5 microL administered in the dorsal raphe on each acquisition training day 5 min before bilateral intrahippocampal injection of 4 microg/microL scopolamine, 8-hydroxy-2 (di-n-propylamino) tetralin (8-OH-DPAT), a 5-HT1A receptor agonist, had no effect on choice accuracy and latency or errors of omission but completely antagonized the impairment of choice accuracy by intrahippocampal scopolamine. Administered into the dorsal raphe at 0.2 and 1 microg/0.5 microL, WAY 100635, a 5-HT1A receptor antagonist, had no effect on rats' performance or on the impairment caused by intrahippocampal scopolamine but dose-dependently antagonized the effect of 1 microg/0.5 microL 8-OH-DPAT on the scopolamine-induced deficit. The results show that stimulation of presynaptic 5-HT1A receptors in the dorsal raphe reverses the deficit caused by intrahippocampal scopolamine, probably by facilitating the transfer of facilitatory information from the entorhinal cortex to the hippocampus. Together with a previous study showing that blockade of postsynaptic hippocampal 5-HT1A receptors antagonized the effect of intrahippocampal scopolamine in the two-platform spatial discrimination task (Carli et al., 1995b), the results suggest that drugs with presynaptic stimulatory and postsynaptic blocking actions on 5-HT1A receptors, such as partial agonists at these receptors, may be useful in the symptomatic treatment of human memory disturbances associated with loss of cholinergic innervation to the hippocampus. PMID- 9753131 TI - Functional GABA(A) receptors on human glioma cells. AB - Glioma cells in acute slices and in primary culture, and glioma-derived human cell lines were screened for the presence of functional GABA(A) receptors. Currents were measured in whole-cell voltage clamp in response to gamma aminobutyric acid (GABA). While cells from the most malignant glioma, the glioblastoma multiforme, did not respond to GABA, an inward current (under our experimental conditions with high Cl- concentration in the pipette) was induced in gliomas of lower grades, namely in 71% of oligodendroglioma cells and in 62% of the astrocytoma cells. Glioma cell lines did not express functional GABA(A) receptors, irrespective of the malignancy of the tumour they originate from. The currents elicited by application of GABA were due to activation of GABA(A) receptors; the specific agonist muscimol mimicked the response, the antagonists bicuculline and picrotoxin blocked the GABA-activated current and the benzodiazepine receptor agonist flunitrazepam augmented the GABA-induced current and the benzodiazepine inverse agonist DMCM decreased the GABA current. Cells were heterogeneous with respect to the direction of the current flow as tested in gramicidin perforated patches: in some cells GABA hyperpolarized the membrane, while in the majority it triggered a depolarization. Moreover, GABA triggered an increase in [Ca2+]i in the majority of the tumour cells due to the activation of Ca2+ channels. Our results suggest a link between the expression of GABA receptors and the growth of glioma cells as the disappearance of functional GABA(A) receptors parallels unlimited growth typical for malignant tumours and immortal cell lines. PMID- 9753132 TI - Reduced adenosine uptake accelerates ischaemic block of population spikes in hippocampal slices from streptozotocin-treated diabetic rats. AB - We have used rats with streptozotocin-induced diabetes to investigate the effects of hyperglycaemia-mediated impaired nucleoside uptake on the actions of endogenous adenosine in hippocampal slices. In control tissue under conditions of anoxia and aglycaemia the rise in the extracellular adenosine concentration resulted in complete inhibition of synaptic activity in about 2 min. In slices from previously hyperglycaemic rats the inhibition of synaptically mediated responses occurred significantly faster, although this change could be prevented by insulin treatment. Application of the selective adenosine A1 receptor antagonist [8-cyclopentyl-1,3-dipropylxanthine (DPCPX)] prevented the anoxia/aglycaemia-mediated inhibition and, furthermore, abolished the differences in the electrophysiological responses between control and diabetic tissue. The effects of impaired nucleoside uptake could be mimicked in control slices by applying the nucleoside uptake blocker hydroxynitrobenzylthioinosine (HNBTI). This had the effect of speeding up the rate of anoxia/aglycaemia-induced synaptic inhibition in control tissue to that seen in diabetic tissue. However, such treatment had no effect on the responses in diabetic tissue as expected if the HNBTI-sensitive uptake process was already inhibited by the chronic hyperglycaemia. The impairment of nucleoside uptake by chronic hyperglycaemia results in the potentiation of the modulatory actions of endogenous adenosine in the central nervous system. Such an alteration in adenosine function may be important in explaining behavioural and pathological changes associated with diabetes mellitus. PMID- 9753133 TI - Induction of tumour-suppressor phosphoprotein p53 in the apoptosis of cultured rat cerebellar neurones triggered by excitatory amino acids. AB - We found that primary cultures of rat cerebellar granule cells, although definitely postmitotic and terminally differentiated, express the tumour suppressor phosphoprotein p53. In particular, granule cells both expressed significant levels of p53 mRNA and positively reacted to an anti-p53 antibody, from the first day of culturing. During neurone differentiation, p53 mRNA content did not significantly change, at least up to 12 days in vitro, while p53 immunoreactivity increased gradually. p53 expression appeared to be further modulable being upregulated after stimulation of glutamate ionotropic receptors by glutamate or kainate. Although qualitatively similar, p53 induction by glutamate and kainate differed in terms of intensity and time-course. The glutamate increase of p53 immunoreactivity appeared within 30 min after the treatment and lasted for at least 2 h. Kainate-induced increase of p53 immunoreactivity was delayed, becoming apparent within 2 h and lasting for at least 8 h. Both kainate- and glutamate-induced increases of p53 immunoreactivity were prevented by the non-competitive NMDA receptor antagonist MK 801. As shown by the electrophoretic mobility shift analysis, both glutamate and kainate induced increases of p53 DNA binding activity. Blockade of p53 induction by a specific p53 antisense oligonucleotide resulted in a partial reduction of excitotoxicity with a complete inhibition of the excitatory amino acids induced apoptosis. Our data suggest that stimulation of ionotropic glutamate receptors in neurones results in a p53-dependent apoptosis. PMID- 9753134 TI - Nongenomic effects of oestrogen: embryonic mouse midbrain neurones respond with a rapid release of calcium from intracellular stores. AB - Evidence is emerging that oestrogen, besides acting via classical nuclear receptors, can rapidly influence the physiology of nerve cells through other mechanisms. Oestrogens have been shown to modulate the differentiation and function of embryonic midbrain dopaminergic neurones by stimulating neurite outgrowth, expression of tyrosine hydroxylase mRNA, dopamine uptake and release in spite of the fact that dopaminergic cells in the prenatal midbrain do not express the classical oestrogen receptor. This study therefore intended to unravel possible signal transduction pathways activated by oestrogen which might be associated with the above oestrogen effects. As a physiological second messenger mechanism, we studied the influence of oestrogen on fluctuations of intracellular Ca2+ levels [Ca2+]i by microspectrofluorimetry of the Ca2+ sensitive indicator Fura-2, in primary cultures from embryonic mouse midbrains. 17Beta-estradiol (10 nM-1 pM) but not 17alpha-estradiol increased [Ca2+]i within 1-3 s in a dose-dependent way. Removal of extracellular Ca2+ abrogated K+ stimulated Ca2+ rise but did not affect 17beta-estradiol stimulation. Pretreatment of cells with thapsigargin (1 microM, 10 min), an inhibitor of Ca2+ pumping ATPases in the endoplasmic reticulum, abolished the 17beta-estradiol effect but not the K+-stimulated [Ca2+]i rise. Oestrogen effects on [Ca2+]i were completely mimicked by using a membrane-impermeant oestrogen-BSA construct. In order to identify oestrogen-sensitive cells, some cultures were subsequently immunostained for microtubule-associated protein II, tyrosine hydroxylase, or GABA. All oestrogen-sensitive cells were immunocytochemically characterized as neurones, and about half of these responsive neurones was found to be dopaminergic or GABAergic. These results demonstrate that 17beta-estradiol is capable of rapidly modulating physiological parameters of developing midbrain neurones by directly interacting with specific membrane binding sites coupled to a signal transduction mechanism that causes a calcium release from intracellular Ca2+ stores. It is suggested that oestrogen effects on differentiation and function of midbrain dopaminergic neurones are mediated by intracellular Ca2+ signalling. PMID- 9753135 TI - In utero gene transfer reveals survival effects of nerve growth factor on rat brain cholinergic neurones during development. AB - Nerve growth factor (NGF) is a maintenance factor for cholinergic neurones in the brain, but its properties as a developmental survival factor are largely unknown. The low accessibility of the developing mammalian brain to experimental manipulation makes it difficult to increase NGF levels during the early phases of brain development. In the present study we have used an in utero, ex-vivo gene transfer approach to explore NGF actions during development of the cholinergic system in the rat brain. Significantly increased numbers of cholinergic neurones were found only in the mesopontine complex in animals receiving NGF-secreting transplants, whereas the cholinergic neurones in the basal forebrain and striatum were not clearly affected. The present results suggest that overexpression of NGF during development may promote the survival of distinct populations of central cholinergic neurones into adulthood. PMID- 9753136 TI - VIP and PACAP potentiate the action of glutamate on BDNF expression in mouse cortical neurones. AB - In view of the neurotrophic effect of vasoactive intestinal peptide (VIP), the regulation of brain-derived neurotrophic factor (BDNF) expression by VIP and the related peptide pituitary adenylate cyclase-activating polypeptide (PACAP) was analysed by Northern blot in primary cultures of cortical neurones. Results reported in this article demonstrate that VIP and PACAP stimulate the expression of BDNF mRNA in primary cultures of cortical neurones and astrocytes. In primary cultures of cortical neurones, induction of BDNF mRNA by VIP and PACAP is completely inhibited by the N-methyl-D-aspartate (NMDA) receptor antagonists MK 801 and AP5, therefore indicating that VIP and PACAP do not stimulate BDNF expression directly but rather by potentiating the effect of glutamate tonically released by neurones and acting at NMDA receptors. In addition to its neurotrophic effects, BDNF has been shown to be involved in neuronal plasticity and results reported here suggest that by stimulating BDNF expression, VIP and PACAP could modulate synaptic plasticity in the cerebral cortex. PMID- 9753137 TI - Pre- and postsynaptic blockade of neuromuscular transmission by Miller-Fisher syndrome IgG at mouse motor nerve terminals. AB - Miller-Fisher syndrome, a variant of an acute inflammatory neuropathy is often associated with serum antibodies to the ganglioside GQ1b, but the pathogenic role of these antibodies and other serum factors is unclear. We here investigated the effect of highly purified immunoglobulin G (IgG) from patients with typical Miller-Fisher syndrome, recording quantal endplate currents by means of a perfused macro-patch-clamp electrode on hemidiaphragms of adult mice. The GQ1b positive and the GQ1b-negative Miller-Fisher IgG as well as its monovalent Fab fragments depressed evoked quantal release in a fast and fully reversible, concentration and voltage dependent manner. The time-course of quantal release was changed with the late releases becoming more frequent. The extent of depression of release followed a Michaelis-Menten kinetic and depended on the extracellular calcium concentration. In addition the amplitude of quanta was reduced postsynaptically. IgG and sera from healthy subjects had no effect. Our results indicate that in Miller-Fisher syndrome, IgG antibodies to an undetermined antigen depress the release process, most likely by interfering with the presynaptic Ca2+ inflow or by interacting with proteins of the exocytotic apparatus, and prevent the activation of postsynaptic channels. Antibodies thus seem to be one pathogenic factor for muscle weakness in Miller-Fisher syndrome and our findings may explain why muscle strength recovers rapidly after therapeutical plasmapheresis. PMID- 9753138 TI - In vitro and in vivo behaviour of NDF-expanded monkey Schwann cells. AB - Schwann cells, the myelin-forming cells of the peripheral nervous system may play a major role in the regeneration and remyelination not only of the peripheral but also of the central nervous system. The discovery of the mitogenicity of human recombinant forms of neuregulins (glial growth factors) on primate Schwann cells allows us to envisage a considerable expansion of these cells in culture with a view to autologous transplantation in the central nervous system. To assay this possibility, we used human recombinant neu-differentiation factor beta (NDFbeta) to expand monkey Schwann cells derived from perinatal and adult nerve biopsies. We report that NDFbeta containing the epidermal growth factor (EGF)-like domain (residues 177-228) is a potent mitogen for monkey Schwann cells but is more effective on perinatal than adult Schwann cells. Moreover, continuous treatment with NDFbeta, does not seem to prevent Schwann cells differentiation into myelin forming cells after their transplantation into the demyelinated mouse spinal cord. These observations, in addition to the close similarities of in vitro behaviour which exist between human and monkey Schwann cells, indicate that monkey Schwann cells could be an ideal tool to study the potential and limits of autologous transplantation in a non-human primate model of central nervous system demyelination. PMID- 9753139 TI - VAMP-1 and VAMP-2 gene expression in rat spinal motoneurones: differential regulation after neuronal injury. AB - Vesicle-associated membrane protein (VAMP; synaptobrevin) is involved in the molecular regulation of transmitter release at the presynaptic plasma membrane. VAMP exists in two isoforms, VAMP-1 and VAMP-2, which are transcribed from two separate genes and differentially expressed in the nervous system. In situ hybridization was used to examine whether VAMP isoform mRNA expression may be altered by experimental manipulations. The effect of nerve injury on VAMP-1 and VAMP-2 mRNA levels in motoneurones of the rat lumbar spinal cord was compared with lesion-induced changes in the expression of choline acetyl transferase (ChAT) and alpha-calcitonin gene-related peptide (alpha-CGRP) mRNA. After unilateral sciatic nerve transection (axotomy), VAMP-1 mRNA expression decreased significantly in parallel with a downregulation of ChAT mRNA in axotomized motoneurones compared with the corresponding motoneurones on the contralateral unlesioned side. There was a rapid decrease in VAMP-1 and ChAT mRNA levels at 2 days after axotomy, and at 7 days there was a 65% decrease in VAMP-1 mRNA and a 48% decrease in ChAT mRNA. VAMP-1 mRNA levels continued to decrease at 14 and 21 days, while ChAT mRNA levels had returned to normal at this time. In contrast, VAMP-2 and alpha-CGRP mRNA levels were upregulated in axotomized motoneurones. A significant increase for both VAMP-2 and alpha-CGRP mRNA levels was present 2 days after axotomy, and a maximum was reached after 7 days for alpha-CGRP mRNA (163%) and after 14 days for VAMP-2 mRNA (587%). Immunohistochemical analysis did not reveal any detectable changes in VAMP-1- or VAMP-2-like immunoreactivity in the motoneurone cell soma after axotomy. In the proximal end of the transected sciatic nerve, there was an increase in VAMP-1- and VAMP-2-LI, which was most prominent at 2 days after lesion. The results show that, in axotomized spinal motoneurones, VAMP-1 mRNA is downregulated and VAMP-2 mRNA is upregulated, indicating differential regulation of the two separate VAMP genes and differential roles for the two VAMP isoforms in the regulation of exocytosis after nerve injury. PMID- 9753141 TI - GluR1 and GluR2/3 subunits of the AMPA-type glutamate receptor are associated with particular types of neurone in laminae I-III of the spinal dorsal horn of the rat. AB - GluR1 and GluR2 subunits of the alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionate (AMPA) receptor are expressed at high levels by neurones in laminae I-III of rat spinal dorsal horn, an area which contains numerous, densely packed small neurones. In order to determine whether these subunits are expressed by inhibitory or excitatory neurones, we combined pre-embedding immunocytochemistry with antibodies that recognize either GluR1, or an epitope common to GluR2 and 3, with postembedding detection of gamma-aminobutyric acid (GABA) and glycine. Most (78%) of the neurones with GluR1-immunoreactivity were GABA-immunoreactive, and some of these were also glycine-immunoreactive, whereas nearly all (97%) of the GluR2/3-immunoreactive neurones were not GABA- or glycine immunoreactive. We carried out double-immunofluorescence and confocal microscopy to provide further information on the neurochemistry of cells that express these subunits. As expected, all neurotensin- and virtually all somatostatin immunoreactive cells (which are thought to be excitatory interneurones) were GluR2/3- but not GluR1-immunoreactive, whereas parvalbumin-containing cells (most of which are GABAergic) possessed GluR1-, but usually not GluR2/3 immunoreactivity. Neurones that contained nitric oxide synthase (most of which are GABAergic) were more variable, with 57% GluR1-immunoreactive and 41% GluR2/3 immunoreactive. Cholinergic neurones in lamina III (which are also GABAergic) invariably showed each type of GluR-immunoreactivity. These results suggest that neuronal populations in laminae I-III have characteristic patterns of GluR expression: GluR1 is particularly associated with inhibitory neurones, and GluR2 with excitatory neurones. This makes it likely that some of the AMPA receptors present on the inhibitory interneurones lack the GluR2 subunit, and may therefore have significant Ca2+-permeability. PMID- 9753140 TI - Two types of calcium currents of the mouse bipolar cells recorded in the retinal slice preparation. AB - In the vertebrate retina, the bipolar cell makes reciprocal synapses with amacrine cells at the axon terminal. It has been postulated that amacrine cells may control the transmitter release from bipolar cells by modulating their calcium currents (ICa). To clarify this possibility calcium currents were studied in bipolar cells of the mouse retina using a slice preparation. ICa was identified by voltage clamp protocols, ionic substitution and pharmacological tools. Depolarization to -30 mV from a holding voltage of -80 mV induced an inward current consisting of an initial transient and a long-lasting sustained component. The transient component was inactivated by holding the membrane at more positive voltages. Addition of 100 microM nifedipine suppressed the sustained component, leaving the transient component almost intact. The sustained component was enhanced when external solution contained 0.1 microM Bay K 8644 or when the external Ca2+ was substituted by equimolar Ba2+. Omega-conotoxin (10 microM omega-ctxn GVIA) did not alter either component. We concluded that the transient component is a low-voltage activated T-type ICa, while the sustained component is a high-voltage activated L-type ICa. T-type ICa was recorded in all cells tested, while L-type ICa was found only in cells that retained axon terminals ramifying in the inner plexiform layer. Thus, it is highly likely that L-type ICa is generated at the axon terminal and contributes to the transmitter release from the bipolar cell. The present results confirm that in addition to the T-type ICa that had been previously described, bipolar cells of the mammalian retina also contain L-type ICa similar to the one that has been reported in bipolar cells of the goldfish. The use of retinal slice preparation allowed us to record this current that was not seen previously in the dissociated mouse bipolar cells. PMID- 9753143 TI - Distribution of GABA receptor rho subunit transcripts in the rat brain. AB - The gamma-aminobutyric acid (GABA) receptor rho subunits recently cloned from rat and human retina are thought to form GABA receptor channels belonging to a pharmacologically distinct receptor class, termed GABA(C). In this work we have examined the distribution of rho1, rho2 and rho3 subunits, and found expression of all three transcripts in several regions of the rat nervous system. In situ hybridization revealed expression of rho2 in the adult rat retina and some other parts of the visual pathways. A high local rho2 expression was seen in the superficial grey layer of the superior colliculus, and in the dorsal lateral geniculate nucleus. Expression was also detected in the 6th layer of visual cortex and in the CA1 pyramidal cell layer of hippocampus. With reverse transcriptase-polymerase chain reaction, expression of rho1 was mainly seen in the adult rat retina and dorsal root ganglia, as well as, at a significantly lower level, in the superior colliculus, hippocampus, brain stem, thalamus, postnatal day 8 (P8) superior colliculus and P8 hippocampus. Expression pattern of rho3 mRNA was clearly different from that of rho1 and rho2, being strongest in the hippocampus, and significantly lower in the retina, dorsal root ganglia and cortex. No rho3 expression was observed in adult or P8 superior colliculus or in P8 hippocampus. The present results clearly demonstrate that expression of GABA receptor rho subunits is not restricted to the retina, but significant expression can also be detected in many other brain regions, especially in those belonging to the visual pathways. The expression pattern of the rho subunits may be helpful in solving the functional significance of the receptors formed from these subunits. PMID- 9753142 TI - Dominant GABA(A) receptor/Cl- channel kinetics correlate with the relative expressions of alpha2, alpha3, alpha5 and beta3 subunits in embryonic rat neurones. AB - The embryonic appearance of GABAergic cells and signals in the rat nervous system coincides with the appearance of transcripts encoding some but not all of the subunits forming GABA(A) receptor/Cl- channels. Quantitative in situ hybridization studies reveal higher variabilities in alpha2 and alpha3 subunit transcripts relative to others examined (alpha5, beta2, beta3 and gamma2) in six spinal and supraspinal regions. Immunocytochemistry of cells dissociated from the embryonic CNS shows that alpha2 and alpha3 subunits are detectable in differentiating neurones. FACS analyses of dissociated cells immunostained with alpha2- or alpha3- antibodies reveal immunopositive subpopulations of variable size in each region. Whole-cell recordings of acutely adherent neurones show that GABA activates Cl- currents whose fluctuations characteristically vary depending on a neurone's region of origin. Spectral analyses indicate a predominance of the low frequency (< 5 Hz) components, which vary regionally. Regression analyses reveal that (i) channel properties correlate with subunit transcript levels and (ii) dominant channel kinetics correlate with alpha2 and alpha3 subunit transcripts indexed as a ratio and with coexpressions of alpha5 and beta3. The correlations strongly suggest that alpha3 subunits in embryonic neurones are expressed in native receptor/channel complexes with slower kinetics than those containing alpha2 without alpha3 subunits. Thus, GABA(A) receptor/Cl- channels in these embryonic neurones may be encoded by the six transcripts (alpha2, alpha3, alpha5, and beta2, beta3, and gamma2) with proportions of alpha2, alpha3, alpha5, and beta3 subunits critical in determining their dominant kinetics. PMID- 9753144 TI - Preprodynorphin mRNA-expressing neurones in the rat parabrachial nucleus: subnuclear localization, hypothalamic projections and colocalization with noxious evoked fos-like immunoreactivity. AB - The dorsal lateral subnucleus of the rat pontine parabrachial nucleus is a major target for ascending nociceptive information from the spinal cord. With in situ hybridization histochemistry, using a radiolabelled cRNA probe, we demonstrate that neurones in and near the dorsal lateral subnucleus express preprodynorphin mRNA. The cRNA probe was constructed from a PCR product amplified from rat genomic DNA. Sequencing of the PCR product revealed that it corresponded to the sequence 466-1101 of the rat preprodynorphin gene exon 4. Tract tracing experiments, using injection of cholera toxin subunit B into the hypothalamic median preoptic nucleus, showed a retrograde labelling pattern of neurones in the parabrachial nucleus that was almost identical to that of the preprodynorphin mRNA expressing neurones. Double-labelling, combining immunohistochemical detection of tracer and in situ hybridization, revealed that the retrogradely labelled neurones expressed preprodynorphin mRNA. A similar double-labelling, combining in situ hybridization with immunohistochemical detection of noxious evoked fos following formalin injection into one hindpaw of awake animals, showed that almost all fos-immunoreactive neurones in the dorsal lateral parabrachial subnucleus also expressed preprodynorphin mRNA. Quantitative analysis suggested that the evoked fos immunoreactivity was accompanied by an increased preprodynorphin mRNA expression. The findings provide evidence that neurones in the dorsal lateral subnucleus produce dynorphin and project to the median preoptic nucleus, and that noxious stimulation in awake animals synaptically activates the dynorphinergic neurones in this subnucleus. These observations are consistent with the idea of a functional and chemical heterogeneity among different parabrachial subnuclei that serves to produce specific homeostatic responses to stimuli that changes the physiological status of the organism, including tissue damage. PMID- 9753145 TI - Origin and terminal distribution of the trigeminal projections to the inferior and superior colliculi in the lesser hedgehog tenrec. AB - The trigemino-tectal projections were investigated with anterograde and retrograde tracing techniques in the Madagascan lesser hedgehog tenrec, Echinops telfairi. There were prominent contralateral projections to the inferior colliculus (CoI) and the superior colliculus (CoS), each showing its own characteristic pattern of terminations. While the projections to the CoI were confined consistently to a circumscribed region in its ventrolateral, external portion, the projections from particularly the rostral trigeminal subdivision to the CoS were distributed inhomogenously across almost the entire rostro-caudal and mediolateral extents. Comparing these data with the spino-tectal projections published previously, it demonstrates that the somatotopic organization of ascending tectal afferents is more distinct in the CoI than in the CoS. There were roughly twice as many trigeminal neurones projecting to the CoS than to the CoI. This difference might be due to the fact that the cells projecting to CoS were distributed extensively across the trigeminal nuclear complex (peak densities in the principal and interpolar subdivisions), while the neurones projecting to the CoI were largely confined to the interpolar and caudal trigeminal subdivisions. The latter cells were located adjacent to the spinal trigeminal tract; the neurones projecting to the CoS occupied preferentially the ventral trigeminal regions at rostral levels, while from the interpolar subdivision caudalward the labelled cells shifted dorsolaterally. In comparison to other mammals the trigeminal projection to the tenrec's CoI is unique. There is evidence for such a projection in other species too, but it is poorly documented, presumably due to technical reasons. PMID- 9753146 TI - Attenuation of the seizure-induced expression of BDNF mRNA in adult rat brain by an inhibitor of calcium/calmodulin-dependent protein kinases. AB - We have examined the potential involvement of calcium/calmodulin-dependent protein kinases in the regulation of brain-derived neurotrophic factor mRNA in vivo following kainic acid (kainate)-induced seizure activity by in situ hybridization. KN-62, a specific inhibitor of calcium/calmodulin-dependent protein kinase type II and IV, blocked the characteristic induction of brain derived neurotrophic factor mRNA seen following seizure activity. This blockade was specific to calcium/calmodulin-dependent protein kinase type II and IV as inhibitors of both protein kinase C and cAMP-dependent protein kinase had no effect. Inhibition of brain-derived neurotrophic factor mRNA increases varied between brain regions; an almost complete inhibition was seen throughout cortical regions, whereas only partial inhibitory effects were noted within hippocampus. A similar inhibition of increased c-fos mRNA was observed throughout cortical, hippocampal and diencephalic regions. The two predominant brain-derived neurotrophic factor transcripts induced by kainate, containing exons I or III, were differentially affected by KN-62. The cortical induction of exon I was blocked by KN-62, whereas exon III was not, providing additional evidence for the differential regulation of individual brain-derived neurotrophic factor transcripts and demonstrating that inhibition of brain-derived neurotrophic factor induction was not due to general blockade of seizure activity throughout the neocortex. These data implicate calcium/calmodulin-dependent protein kinase type II or IV in the regulation of brain-derived neurotrophic factor mRNA in vivo and suggest regionally specific mechanisms occur throughout the brain. PMID- 9753147 TI - Possible involvement of gradients in guidance of receptor cell axons towards their target position on the olfactory bulb. AB - There is increasing evidence for directional guidance of growing axons by molecular gradients in target tissues. Aside from biochemical studies on gradients and their role, the capability of axons to approach their target position from different aspects of a two-dimensional field is itself an indication for guidance by gradients. According to this criterion, such guidance is expected to be involved not only in map-formation in the visual system but also in targeting of receptor cell axons in the olfactory bulb. In this paper, physico-chemical concepts of visual mapping are adapted to olfactory targeting. In both cases there must be sophisticated processing of graded cues in the growing tip of the axon for growth cone navigation. In visual map formation, a target position is determined by influences of cues depending on the position of axonal origin; in olfactory targeting, however, these influences are expected to be based on properties of the receptor-cell-specific molecules (possibly including the receptor molecule itself), as well as by gene regulation affecting the levels of expression. According to this concept, the main role of molecules expressed in a receptor-cell-type specific manner is not matching specific counterparts on the target tissue, but instead quantitative modulation of growth cone steering for sensing the direction towards the target position. PMID- 9753148 TI - Prenatal differentiation of mouse vomeronasal neurones. AB - The vomeronasal organ (VNO) subserves basic chemosensory functions in rodents, mainly related to sexual behaviour. In order to understand early stages of the VNO structural maturation, we have undertaken an immunocytochemical analysis of the VNO of fetal mice. Our results demonstrate that Olfactory Marker Protein (OMP), a marker of differentiated chemosensory cells, is already expressed in vomeronasal neurones and their fibres projecting to the accessory olfactory bulb during the last week of gestation. However, in contrast to the adult, where its expression is restricted to the medial sensory neuronal component of the VNO, during fetal development OMP is also present in cells located in the lateral non sensory epithelial component. Some other markers of nasal chemosensory neurones, such as GAP-43/B-50, Protein Gene Product 9.5 (PGP 9.5) and carnosine are also transiently expressed in this ectopic site. These results indicate that (i) significant morphological and biochemical maturation of the VNO is achieved before birth; (ii) transient cell populations, sharing the biochemical profile of the vomeronasal chemosensory receptors, occur in ectopic areas during fetal development. PMID- 9753150 TI - The neural processing of 3-D visual information: evidence from eye movements. AB - Primates have several reflexes that generate eye movements to compensate for bodily movements that would otherwise disturb their gaze and undermine their ability to process visual information. Two vestibulo-ocular reflexes compensate selectively for rotational and translational disturbances of the head, and each has visual backups that operate as negative feedback tracking mechanisms to deal with any residual disturbances of gaze. Of particular interest here are three recently discovered visual tracking mechanisms that specifically address translational disturbances and operate in machine-like fashion with ultra-short latencies (< 60 ms in monkeys, < 85 ms in humans). These visual reflexes deal with motions in all three dimensions and operate as automatic servos, using preattentive parallel processing to provide signals that initiate eye movements before the observer is even aware that there has been a disturbance. This processing is accomplished by visual filters each tuned to a different feature of the binocular images located in the immediate vicinity of the plane of fixation. Two of the reflexes use binocular stereo cues and the third is tuned to particular patterns of optic flow associated with the observer's forward motion. Some stereoanomalous subjects show tracking deficits that can be attributed to a lack of just one subtype of cortical cell encoding motion in one particular direction in a narrow depth plane centred on fixation. Despite their rapid, reflex nature, all three mechanisms rely on cortical processing and evidence from monkeys supports the hypothesis that all are mediated by the medial superior temporal (MST) area of cortex. Remarkably, MST seems to represent the first stage in cortical motion processing at which the visual error signals driving each of the three reflexes are fully elaborated at the level of individual cells. PMID- 9753149 TI - Nitric oxide sensitive depolarization-induced hyperpolarization: a possible role for gap junctions during development. AB - Electrical coupling is a widespread feature of developing neuronal circuits and it contributes to the generation of patterned activity. In the developing rat hippocampus, release of GABA by coactive hilar interneurones generates widespread synchronized activity. Here it is shown that hilar interneurones strongly rectify in the outward direction when depolarized. This depolarization-induced hyperpolarization, abolished by gap junction uncouplers, is modulated by nitric oxide. This phenomenon might represent a current-shunting mechanism of the excess current by providing functional inhibition at a developmental stage when GABA is excitatory. Spatial buffering of the current might represent an osmotic mechanism for growth and differentiation. PMID- 9753151 TI - Characterization of perforant path lesions in rodent models of memory and attention. AB - Early stage Alzheimer's disease (AD) pathology is associated with neurodegeneration of systems within the temporal cortex, e.g. the entorhinal cortex, perforant pathway and hippocampus. The perforant pathway provides the major neuronal input to the hippocampus from the entorhinal cortex and thus relays multimodal sensory information derived from cortical zones into the hippocampus. The earliest symptoms of AD include cognitive impairments, e.g. deficits in short-term memory and attention. Consequently, we have investigated the effect of bilateral knife cut lesions to the perforant path on cognition in rats using models measuring primarily short-term memory (operant delayed match to position task), attention (serial five-choice reaction time task) and spatial learning (Morris water maze). Rats receiving bilateral perforant path lesions showed normal neurological function and a mild hyperactivity. The lesion produced little effect on attention assessed using the five-choice task. In contrast, animals with equivalent lesions showed a robust delay-dependent deficit in the delayed match to position task. Spatial learning in the water maze task was also severely impaired. The delay-dependent deficit in the match to position task was not reversed by tacrine (3 mg/kg) pretreatment. The present data support a selective impairment of cognitive function following perforant path lesions that was confined to mnemonic rather than attentional processing. These findings complement primate and human studies identifying a critical role of the perforant pathway and associated temporal lobe structures in declarative memory. Degeneration of the perforant pathway is likely to contribute to the mnemonic deficits characteristic of early AD. The failure of tacrine to ameliorate these deficits may be relevant to an emerging clinical literature suggesting that cholinomimetic therapies improve attentional rather than mnemonic function in AD. PMID- 9753152 TI - Polarized sorting of nicotinic acetylcholine receptors to the postsynaptic membrane in Torpedo electrocyte. AB - Several regulatory mechanisms contribute to the accumulation and maintenance of high concentrations of acetylcholine receptors (AChR) at the postsynaptic membrane of the neuromuscular junction, including compartmentalized gene transcription, targeting, clustering and anchoring to the cytoskeleton. The targeting of the AChR to the postsynaptic membrane is likely to involve a polarized sorting in the exocytic pathway. In this work, we used the electrocyte of Torpedo marmorata electric organ to study the intracellular trafficking of neosynthesized AChR and its delivery to the postsynaptic membrane. Gradient centrifugation and immunoisolation techniques have led to the isolation of two populations of post-Golgi transport vesicles (PGVs) enriched in proteins of either the innervated (AChR) or non-innervated (Na,K-ATPase) membrane domains of the cell. Immunolabelling of these vesicles at the EM level disclosed that very few PGVs contained both proteins. In AChR-enriched vesicles, high sialylation of AchR molecules, an expected post-translational modification of proteins exiting the trans-Golgi network, and the presence of a marker of the exocytic pathway (Rab6p), indicate that these vesicles are carriers engaged in the Golgi-to-plasma membrane transport. These data suggest that AChR and Na,K-ATPase are sorted intracellularly most likely within the trans-Golgi network. Furthermore, EM analysis and immunogold-labelling experiments provided in situ evidence that the AChR-containing PGVs are conveyed to the postsynaptic membrane, possibly by a microtubule-dependent transport mechanism. Our data therefore provide the first evidence that the targeting of receptors for neurotransmitters to synaptic sites could be contributed by intracellular sorting and polarized delivery in the exocytic pathway. PMID- 9753153 TI - Mechanisms and functional significance of a slow inhibitory potential in neurons of the lateral amygdala. AB - A slow inhibitory potential (sIP) elicited upon synaptic activation in spiny, pyramidal-like cells with properties indicative of projection neurons was investigated in slices of the rat and guinea-pig lateral amygdala in vitro. The sIP succeeded the triphasic sequence of excitatory and fast/slow inhibitory postsynaptic potentials mediated via glutamate and GABA(A/B) receptors, respectively, was readily evoked upon repetitive stimulation of the external capsule and appeared to terminate epileptiform burst discharges during pharmacologically reduced GABAergic influence. The sIP reversed close to the Cl- equilibrium potential, but was not affected by altered transmembrane Cl- gradients and not abolished by antagonists to ligand-gated Cl- channels. Intracellular injection of QX 314 and resulting blockade of sodium spikes had no effect, whereas the Ca2+ chelator BAPTA blocked the sIP concomitantly with slow hyperpolarizing afterpotentials following intrinsically generated spike firing, thereby indicating the contribution of Ca2+-dependent mechanisms secondary to synaptic activation. During action of BAPTA and QX 314, an N-methyl-D-aspartate (NMDA) receptor-mediated potential was unmasked, which contributed to the sIP. The Ca2+-dependent mechanisms of the sIP involved a membrane K+ conductance, as was indicated by the dependence on the K+ gradient and the shift of the reversal potential towards the K+ equilibrium potential during blocked NMDA receptors. During the presence of GABA receptor antagonists, reduction of the Ca2+-activated K+ conductance through injection of BAPTA or application of dopamine induced a gradual shift of interictal-like single bursts of spikes towards the generation of re-occurring ictal-like activity. It is concluded that pyramidal-like projection cells in the AL can generate a sIP upon synaptic activation, which reflects the combined activation of an NMDA receptor-mediated cation current and a K+ current that is secondary to the rise in intracellular Ca2+ concentration resulting from the preceding depolarizing response. The sIP may play an important role in controlling excitatory activity in the amygdala, particularly in preventing the transformation of interictal-like activity towards recurrent epileptic discharges during periods of decreased GABAergic influence. PMID- 9753154 TI - Tumour necrosis factor-alpha and interleukin-2 differentially affect hippocampal serotonergic neurotransmission, behavioural activity, body temperature and hypothalamic-pituitary-adrenocortical axis activity in the rat. AB - Intraperitoneal endotoxin injection and central administration of interleukin (IL)-1beta profoundly activate hippocampal serotonergic neurotransmission. This study was designed to investigate, using in vivo microdialysis, the effects of another endotoxin-induced proinflammatory cytokine, tumour necrosis factor-alpha, and the effects of the non-inflammatory cytokine, IL-2, on hippocampal extracellular levels of serotonin. To compare the effects of these cytokines on neurotransmission with the effects on physiological parameters and behaviour, hypothalamic-pituitary-adrenocortical (HPA) axis activity, body temperature and behavioural activity were monitored as well. Time-dependent changes in serotonergic neurotransmission and HPA axis activity were determined by measuring serotonin, its metabolite 5-hydroxyindoleacetic acid and free corticosterone in dialysates. Total behavioural activity was scored by assessing the time during which rats were active. Core body temperature was measured by biotelemetry. Intracerebroventricular injection of 50 or 100 ng recombinant murine tumour necrosis factor-alpha exerted no effect on hippocampal serotonergic neurotransmission, and induced no signs of sickness behaviour. However, these doses produced a dose-dependent increase in body temperature and free corticosterone levels. In contrast, intracerebroventricular administration of 500 ng, but not of 50 ng, recombinant human IL-2 produced a marked increase in hippocampal extracellular concentrations of serotonin and 5-hydroxyindoleacetic acid, accompanied by a pronounced behavioural inhibition and other signs of sickness. Moreover, both doses of IL-2 caused a dose-dependent increase in body temperature and free corticosterone levels. Interestingly, intracerebroventricular pretreatment with the IL-1 receptor antagonist showed that the effects of IL-2 on hippocampal serotonin were completely dependent on endogenous brain IL-1. However, IL-1 seemed to play only a minor role in the IL-2 induced increase in free corticosterone. Taken together, the results show that cytokines produce partially overlapping brain-mediated responses, but are selectively effective in stimulating hippocampal serotonergic neurotransmission and inducing sickness behaviour. Moreover, we postulate that activation of hippocampal serotonin release is instrumental in the full development of behavioural inhibition. PMID- 9753155 TI - Characterization of a nicotinic acetylcholine receptor from the insect Manduca sexta. AB - Manduca sexta is a nicotine-insensitive insect, the larval form of which feeds on tobacco. It has been postulated that its nicotine insensitivity may reflect the presence of a modified nicotinic acetylcholine receptor whose alpha subunits lack the amino acid residues necessary for binding nicotine: we have performed ligand binding assays and molecular cloning to examine this hypothesis. [125I]alpha bungarotoxin bound specifically to both larval and adult membranes, with Kd values of 7.6 and 6.5 nM and Bmax values of 119 and 815 fmol/mg protein, respectively. The pharmacological profile of [1251]alpha-bungarotoxin binding was similar in both tissues. In particular, nicotine (Ki values: 1.6 microM and 2 microM for larvae and adults, respectively) competed with an affinity similar to that found for nicotine-sensitive insects. No alpha-bungarotoxin-insensitive binding sites labelled by [3H]epibatidine could be detected. Using the alpha-like subunit from the locust Schistocerca gregaria to probe two cDNA libraries, and by inverse PCR on circularized genomic DNA from Manduca sexta, we have obtained overlapping cDNA clones that contain the complete coding sequence of a putative nicotinic subunit from Manduca sexta (MARA1). No other alpha-subunit cDNAs were isolated using this probe, although it hybridized to multiple bands on Southern blots. The sequence of MARA1 is consistent with an alpha-like subunit capable of binding alpha-bungarotoxin, and it retains all those amino acids implicated in nicotine binding to vertebrate nicotinic receptors. Taken together, these findings provide no support for the hypothesis that the nicotine insensitivity of Manduca sexta is the result of a nicotinic receptor with diminished nicotine binding. PMID- 9753156 TI - Reciprocal modulation of TrkA and p75NTR affinity states is mediated by direct receptor interactions. AB - Equilibrium binding of 125I-nerve growth factor (125I-NGF) to cells coexpressing the tyrosine kinase receptor A (TrkA) and common neurotrophin receptor (p75NTR), cells coexpressing both receptors where p75NTR is occupied, and cells expressing only p75NTR, revealed reciprocal modulation of receptor affinity states. Analysis of receptor affinity states in PC12 cells, PC12 cells in the presence of brain derived neurotrophic factor (BDNF), and PC12nnr5 cells suggested that liganded and unliganded p75NTR induce a higher affinity state within TrkA, while TrkA induces a lower affinity state within p75NTR. These data are consistent with receptor allosterism, and prompted a search for TrkA/p75NTR complexes in the absence of NGF. Chemical crosslinking studies revealed high molecular weight receptor complexes that specifically bound 125I-NGF, and were immunoprecipitated by antibodies to both receptors. The heteroreceptor complex of TrkA and p75NTR alters conformation and/or dissociates in the presence of NGF, as indicated by the ability of low concentrations of NGF to prevent heteroreceptor crosslinking. These data suggest a new model of receptor interaction, whereby structural changes within a heteroreceptor complex are induced by ligand binding. PMID- 9753157 TI - Origin of the synchronized network activity in the rabbit developing hippocampus. AB - Rhythmic spontaneous bursting is a fundamental hallmark of the immature hippocampal activity recorded in vitro. These bursts or giant depolarizing potentials (GDPs) are GABA- and glutamatergic-driven events. The mechanisms of GDPs generation are still controversial, since although a hilar origin has been suggested, GDPs were also recorded from isolated CA3 area. Here, we have investigated the origin of GDPs in hippocampal slices from newborn rabbits. Simultaneous intracellular recordings were performed in CA3, CA1 and the fascia dentata. We found a high degree of correlation between the spontaneous GDPs present in CA3 and CA1 regions. Cross-correlation analysis demonstrated that CA3 firing precedes CA1 by about 192 ms, although a significant population of discharges was recorded first in CA1 (20%). Granule cells (GCs) in the fascia dentata also showed GDPs. The frequency of these events (1.46 +/- 1.25 GDPs/min, n = 7) is significantly lower when compared with that from CA3 (3.13 +/- 1.43 GDPs/min, n = 10) or CA1 (2.94 +/- 1.36 GDPs/min, n = 17). Dual recordings from CA3 and fascia dentata cells showed synchronous bursts in both regions with no prevalent preceding area. By recording from isolated areas we found that CA1, CA3 and the fascia dentata can produce GDPs, suggesting that they emerge as a property of local circuits present throughout the hippocampus. PMID- 9753158 TI - Expression of small-conductance calcium-activated potassium channels (SK) in outer hair cells of the rat cochlea. AB - Physiological evidence suggests that SK-type Ca2+-activated K+ channels participate in ACh-induced hyperpolarization of OHCs (outer hair cells). Based on the sequences published by Kohler et al. [(1996), Science, 273: 1709), we designed degenerated primers recognizing cDNA subunits of rSK1, rSK2 and rSK3. Using this consensus set of primers, we probed by PCR a rat organ of Corti cDNA library. Two PCR products of 707 base pairs with sequence identical to rSK3 and rSK2 were obtained and cloned to generate RNA probes for in situ hybridization in the rat cochlea. The subunit rSK2 showed hybridization in the organ of Corti, at the location of the OHCs. The expression of rSK2 by OHCs was confirmed by probing with PCR a poly(A) amplified OHC cDNA library. During development, rSK2 hybridization in the organ of Corti was negative at embryonic days E16, E18 and at P0, weak at P4 and stronger from P8 to adulthood. The subunit rSK2 could also be detected in the spiral ganglion from P4 to the adult stage. Contrary to rSK2, the subunit rSK3 did not show specific hybridization in the organ of Corti at the adult stage (P120) and only a weak expression was observed at P10 and P21. Our study demonstrates expression of rSK2 in OHCs. These potassium channels are good candidates to underlie the ACh-activated K+ currents recorded during patch-clamp recordings in isolated OHCs. The expression of rSK2 in the cochlear ganglion at the adult stage suggests that SK Ca2+-activated K+ channels may also participate in the repolarization of the auditory neurons after the action potential and may influence their firing patterns. PMID- 9753160 TI - Spectral envelope coding in cat primary auditory cortex: linear and non-linear effects of stimulus characteristics. AB - Electrophysiological studies in mammal primary auditory cortex have demonstrated neuronal tuning and cortical spatial organization based upon spectral and temporal qualities of the stimulus including: its frequency, intensity, amplitude modulation and frequency modulation. Although communication and other behaviourally relevant sounds are usually complex, most response characterizations have used tonal stimuli. To better understand the mechanisms necessary to process complex sounds, we investigated neuronal responses to a specific class of broadband stimuli, auditory gratings or ripple stimuli, and compared the responses with single tone responses. Ripple stimuli consisted of 150-200 frequency components with the intensity of each component adjusted such that the envelope of the frequency spectrum is sinusoidal. It has been demonstrated that neurons are tuned to specific characteristics of those ripple stimulus including the intensity, the spacing of the peaks, and the location of the peaks and valleys (C. E. Schreiner and B. M. Calhoun, Auditory Neurosci., 1994; 1: 39-61). Although previous results showed that neuronal response strength varied with the intensity and the fundamental frequency of the stimulus, it is shown here that the relative response to different ripple spacings remains essentially constant with changes in the intensity and the fundamental frequency. These findings support a close relationship between pure-tone receptive fields and ripple transfer functions. However, variations of other stimulus characteristics, such as spectral modulation depth, result in non-linear alterations in the ripple transformation. The processing between the basilar membrane and the primary auditory cortex of broadband stimuli appears generally to be non-linear, although specific stimulus qualities, including the phase of the spectral envelope, are processed in a nearly linear manner. PMID- 9753159 TI - Structural diversity of the voltage-dependent Ca2+ channel alpha1E-subunit. AB - Voltage-operated Ca2+ channels are heteromultimeric proteins. Their structural diversity is caused by several genes encoding homologous subunits and by alternative splicing of single transcripts. Isoforms of alpha1 subunits, which contain the ion conducting pore, have been deduced from each of the six cDNA sequences cloned so far from different species. The isoforms predicted for the alpha1E subunit are structurally related to the primary sequence of the amino terminus, the centre of the subunit (II-III loop), and the carboxy terminus. Mouse and human alpha1E transcripts have been analysed by reverse transcription polymerase chain reaction and by sequencing of amplified fragments. For the II III loop three different alpha1E cDNA fragments are amplified from mouse and human brain, showing that isoforms originally predicted from sequence alignment of different species are expressed in a single one. Both predicted alpha1E cDNA fragments of the carboxy terminus are identified in vivo. Two different alpha1E constructs, referring to the major structural difference in the carboxy terminus, were stably transfected in HEK293 cells. The biophysical properties of these cells were compared in order to evaluate the importance in vitro of the carboxy terminal insertion found in vivo. The wild-type alpha1E subunit showed properties, typical for a high-voltage activated Ca2+ channel. The deletion of 43 amino acid residues at the carboxy terminus does not cause significant differences in the current density and the basic biophysical properties. However, a functional difference is suggested, as in embryonic stem cells, differentiated in vitro to neuronal cells, the pattern of transcripts indicative for different alpha1E isoforms changes during development. In human cerebellum the longer alpha1E isoform is expressed predominantly. Although, it has not been possible to assign functional differences to the two alpha1E constructs tested in vitro, the expression pattern of the structurally related isoforms may have functional importance in vivo. PMID- 9753161 TI - Glutamate-induced Co2+ uptake in rat auditory brainstem neurons reveals developmental changes in Ca2+ permeability of glutamate receptors. AB - Ca2+ influx through glutamate receptors (GluRs) is thought to play a crucial part in developmental processes and neuronal plasticity. Here we have examined the spatiotemporal distribution of Ca2+-permeable GluRs in auditory brainstem neurons of the rat from birth to adulthood, using the cobalt-staining technique of Pruss and collaborators. In slices of young adult rats, 1 mM glutamate evoked intense cobalt uptake in subsets of neurons in the ventral cochlear nuclei, the medial nucleus of the trapezoid body, the lateral and the medial superior olive, and the lateral lemniscal nuclei. Neurones in the central nucleus of the inferior colliculus, and thalamic auditory nuclei appear to express few, if any, Ca2+ permeable GluRs. Thus, in adults, Ca2+-permeable GluRs are present in neurons of almost all main relay stations of the auditory brainstem. During development, cobalt-stained cells first appeared at about hearing onset (at postnatal day 12 [P12]). At P16, staining levels were highest and the pattern of distribution was already adult-like. The staining intensity slightly declined during the fourth postnatal week. In contrast, Ca2+-permeable receptors were detected in the external cortex of the inferior colliculus as early as P4. Our results show that auditory neurons, characterized by a high temporal precision in neuronal activity, display Ca2+-permeable GluRs. Because Ca2+ permeability appears at about the onset of hearing and is highest during the following 2 weeks, Ca2+ influx through GluRs is likely to be implicated in remodelling processes occurring during this ontogenetic period. PMID- 9753162 TI - Different patterns of induction of FGF-2, FGF-1 and BDNF mRNAs during kindling epileptogenesis in the rat. AB - Neurotrophic factors (NTF) play important roles in the developing and in the adult brain. NTF involvement in neuronal plasticity is suggested by the modulation of NTF expression patterns in different physiological and pathological situations and by the effects they produce in the adult brain (e.g. axonal sprouting induction and neuroprotection). We used the RNAase protection assay to investigate the expression patterns of some NTFs during amygdala kindling, an animal model of epilepsy in which 'pathological' neuronal plasticity appears to occur. After a single kindling stimulation, fibroblast growth factor-2 (FGF-2) mRNA levels were increased in the hippocampus, the cortex and the hypothalamus, whereas they were not significantly altered in the thalamus and the striatum. A single stimulation did not alter fibroblast growth factor-1 (FGF-1) and brain derived neurotrophic factor (BDNF) gene expression. Fully kindled animals, left unstimulated for a week, did not exhibit any alteration in the mRNA levels for any of the NTFs examined. However, in contrast with the effect of a single stimulation, amygdala stimulation of kindled animals (evoking a generalized tonic clonic seizure) produced a great increase in hippocampal and cortical BDNF mRNA levels, but FGF-1 mRNA levels were not altered, and FGF-2 mRNA levels were significantly increased only in the cortex. These results suggest that different NTFs can be recruited at different stages of kindling epileptogenesis and, accordingly, may play different parts in the adaptive changes taking place in this experimental paradigm. PMID- 9753163 TI - Electrophysiological evidence that noradrenergic neurons of the rat locus coeruleus are tonically inhibited by GABA during sleep. AB - It is well known that noradrenergic locus coeruleus (LC) neurons decrease their activity during slow wave sleep (SWS) and are virtually quiescent during paradoxical sleep (PS). It has been proposed that a GABAergic input could be directly responsible for this sleep-dependent neuronal inactivation. To test this hypothesis, we used a new method combining polygraphic recordings, microiontophoresis and single-unit extracellular recordings in unanaesthetized head-restrained rats. We found that iontophoretic application of bicuculline, a specific GABA(A)-receptor antagonist, during PS and SWS restore a tonic firing in the LC noradrenergic neurons. We further observed that the application of bicuculline during wakefulness (W) induced an increase of the discharge rate. Of particular importance for the interpretation of these results, using the microdialysis technique, Nitz and Siegel (Neuroscience, 1997; 78: 795) recently found an increase of the GABA release in the cat LC during SWS and PS as compared with waking values. Based on these and our results, we therefore propose that during W, the LC cells are under a GABAergic inhibitory tone which progressively increases at the entrance and during SWS and PS and is responsible for the inactivation of these neurons during these states. PMID- 9753164 TI - Potassium depolarization of mammalian vestibular sensory cells increases [Ca2+]i through voltage-sensitive calcium channels. AB - The existence of voltage-sensitive Ca2+ channels in type I vestibular hair cells of mammals has not been conclusively proven. Furthermore, Ca2+ channels present in type II vestibular hair cells of mammals have not been pharmacologically identified. Fura-2 fluorescence was used to estimate, in both cell types, intracellular Ca2+ concentration ([Ca2+]i) variations induced by K+ depolarization and modified by specific Ca2+ channel agonists and antagonists. At rest, [Ca2+]i was 90 +/- 20 nM in both cell types. Microperifusion of high-K+ solution (50 mM) for 1 s increased [Ca2+]i to 290 +/- 50 nM in type I (n = 20) and to 440 +/- 50 nM in type II cells (n = 10). In Ca2+-free medium, K+ did not alter [Ca2+]i. The specific L-type Ca2+ channel agonist, Bay K, and antagonist, nitrendipine, modified in a dose-dependent manner the K+-induced [Ca2+]i increase in both cell types with maximum effect at 2 microM and 400 nM, respectively. Ni2+, a T-type Ca2+ channel blocker, reduced K+-evoked Ca2+ responses in a dose dependent manner. For elevated Ni2+ concentrations, the response was differently affected by Ni2+ alone, or combined to nitrendipine (500 nM). In optimal conditions, nitrendipine and Ni2+ strongly depressed by 95% the [Ca2+]i increases. By contrast, neither omega-agatoxin IVA (1 microM), a specific P- and Q-type blocker, nor omega-conotoxin GVIA (1 microM), a specific N-type blocker, affected K+-evoked Ca2+i responses. These results provide the first direct evidence that L- and probably T-type channels control the K+-induced Ca2+ influx in both types of sensory cells. PMID- 9753165 TI - Motor discoordination and increased susceptibility to cerebellar injury in GLAST mutant mice. AB - To study the function of GLAST, a glutamate transporter highly expressed in the cerebellar Bergmann astrocytes, the mouse GLAST gene was inactivated. GLAST deficient mice developed normally and could manage simple coordinated tasks, such as staying on a stationary or a slowly rotating rod, but failed more challenging task such as staying on a quickly rotating rod. Electrophysiological examination revealed that Purkinje cells in the mutant mice remained to be multiply innervated by climbing fibres even at the adult stage. We also found that oedema volumes in the mutant mice increased significantly after cerebellar injury. These results indicate that GLAST plays active roles both in the cerebellar climbing fibre synapse formation and in preventing excitotoxic cerebellar damage after acute brain injury. PMID- 9753166 TI - Regulation of cell-type specific expression of lacZ by the 5'-flanking region of mouse GAD67 gene in the central nervous system of transgenic mice. AB - The transcriptional regulation of the murine gene encoding the 67-kDa form of glutamic acid decarboxylase (GAD67) was studied by beta-galactosidase histochemistry in transgenic mice carrying fusion genes between progressively longer portions of the 5'-upstream regulatory region of GAD67 and E. coli lacZ. No expression was detected in brains of mice carrying 1.3 kb of upstream sequences including a housekeeping and two conventional promoters, and two negative regulatory elements with homology to known silencers. In mice carrying the same portion of the promoter region plus the first intron, lacZ expression in the adult central nervous system was found in few, exclusively neuronal sites. The number of correctly stained GABAergic centres increased dramatically with increasing the length of the 5'-upstream region included in the construct which suggests that multiple putative spatial enhancers are located in this region. Their action is influenced by epigenetic mechanisms that may be due to site-of integration and transgene copy-number effects. Additional cis-acting elements are needed to obtain fully correct expression in all GABAergic neurons of the adult central nervous system. PMID- 9753167 TI - Different cell surface areas of polarized radial glia having opposite effects on axonal outgrowth. AB - During neuronal development neurites are likely to be specifically guided to their targets. Within the chicken retina, ganglion cell axons are extended exclusively into the optic fibre layer, but not into the outer retina. We investigated, whether radial glial cells having endfeet at the optic fibre layer and somata in the outer retina, might be involved in neurite guidance. In order to analyse distinct cell surface areas, endfeet and somata of these glial cells were purified. Glial endfeet were isolated from flat mounted retina by a specific detachment procedure. Glial somata were purified by negative selection using a monoclonal antibody/complement mediated cytolysis of all non-glial cells. Retinal tissue strips were explanted either onto pure glial endfeet or onto glial somata. As revealed by scanning and fluorescence microscopy, essentially no ganglion cell axons were evident on glial somata, whereas axonal outgrowth was abundant on glial endfeet. However, when glial somata were heat treated and employed thereafter as the substratum, axon extension was significantly increased. Time lapse video recording studies indicated that purified cell membranes of glial somata but not of endfeet induced collapse of growth cones. Collapsing activity was destroyed by heat treatment of glial membranes. The collapsing activity of retinal glia was found to be specific for retinal ganglion cell neurites, because growth cones from dorsal root ganglia remained unaffected. Employing four different kinase inhibitors revealed that the investigated protein kinase types were unlikely to be involved in the collapse reaction. The data show for the first time that radial glial cells are functionally polarized having permissive endfeet and inhibitory somata with regard to outgrowing axons. This finding underscores the pivotal role of radial glia in structuring developing nervous systems. PMID- 9753168 TI - The perforant path projection from the medial entorhinal cortex layer III to the subiculum in the rat combined hippocampal-entorhinal cortex slice. AB - Intracellular recordings were performed to examine the perforant path projection from layer III of the entorhinal cortex to the subiculum in rat combined hippocampal-entorhinal cortex slices. Electrical stimulation in the medial entorhinal cortex layer III caused short latency combined excitatory and inhibitory synaptic responses in subicular cells. In the presence of the GABA(A) antagonist bicuculline and the GABA(B) antagonist CGP-55845 A inhibition was blocked and isolated AMPA- or NMDA receptor-mediated EPSPs could be elicited. After application of the non-NMDA antagonist NBQX and the NMDA antagonist APV excitatory responses were completely blocked indicating a glutamatergic input from the neurons of the medial entorhinal cortex layer III. By stimulation from a close (< 0.2 mm) position in the presence of NBQX and APV and either CGP-55845 A or bicuculline we could record monosynaptic fast GABA(A) or slow GABA(B) receptor mediated IPSPs, respectively. We compared synaptic responses in subicular cells induced by stimulation in the medial entorhinal cortex layer III with responses elicited by stimulation of afferent fibres in the alveus. The EPSPs of subicular cells induced by stimulation of alvear fibres could be significantly augmented by simultaneous activation of perforant path fibres originating in the medial entorhinal cortex layer III, while delayed activation of alvear fibres after stimulation of the perforant path resulted in a weak inhibition of the alveus evoked EPSPs. Thus, the perforant path projection activates monosynaptic excitation of subicular neurons. Therefore the entorhinal cortex does not only function as an important input structure of the hippocampal formation but is also able to modulate the hippocampal output via the entorhinal-subicular circuit. PMID- 9753169 TI - Dissociations in dopamine release in medial prefrontal cortex and ventral striatum during the acquisition and extinction of classical aversive conditioning in the rat. AB - Dual perfusion in vivo brain microdialysis was used to monitor extracellular levels of dopamine in the medial prefrontal cortex and ventral striatum during the acquisition and extinction of a classical aversive conditioning paradigm in rats. The main finding was a dissociation in the pattern of release in the two brain areas. The first stimulus-footshock pairing elicited large increases in cortical dopamine over baseline levels that were much greater than the increases elicited by different stimuli of equivalent salience that were unpaired with footshock. In contrast, dopamine levels in ventral striatum were unchanged under these conditions. Over the next two pairings, there was a decline in the cortical response and an increase in the response in ventral striatum. The first presentation of the aversive conditioned stimulus in a separate context elicited the largest response in ventral striatum. Post-conditioning, the cortical response to the conditioned stimulus was smaller than that elicited by the initial stimulus-footshock pairing and was equivalent in magnitude to that elicited by stimuli unpaired with footshock. Over the final two conditioned stimuli presentations, in the absence of the footshock reinforcer (extinction), responses declined in both brain areas. Simultaneous monitoring of behaviour indicated that the neurochemical events were accompanied by effective aversive learning, as indexed by conditioned freezing responses. The data are discussed in terms of the hypothesis that medial prefrontal cortex is especially engaged during novel circumstances which may, potentially, require new learning, whilst ventral striatal dopamine more closely follows the expression of conditioned responding during learning and extinction. PMID- 9753170 TI - The onset of parvalbumin-expression in interneurons of the rat parietal cortex depends upon extrinsic factor(s). AB - Parvalbumin (PV) belongs to the large family of EF-hand calcium-binding proteins and is an excellent marker for a subpopulation of GABAergic neocortical interneurons. During cortical development, PV first appears on postnatal day (P)8, in the infragranular layers; after P14, it also becomes apparent within the supragranular layers. However, nothing is known about the factors controlling its expression, which could involve functional activity, neuronal connectivity and/or neurotrophic factors. It being difficult to manipulate these parameters in vivo, their role may be more readily assessed in organotypic cultures, which are deprived of their subcortical afferents and efferents, and hence of subcortically derived neurotrophic factors and extrinsic functional activity. We prepared slices of the rat brain on P3, P5, P7 and P9, maintained them in culture for 2-5 weeks, and compared the temporal and spatial distribution pattern of PV immunoreactivity within these slices with the in vivo situation. We found, first, that during late postnatal in vivo development and ageing, the number of PV immunoreactive neurons in the parietal cortex decreases significantly, and second, that the expression of PV-immunoreactivity in the parietal cortex was markedly influenced by the phase of postnatal development at which slice cultures were explanted. In those removed on P7 and P9, the number of PV-immunoreactive cells, as well as the temporal and spatial distribution pattern of PV immunoreactivity corresponded to the in vivo situation, but in explants obtained on P3 or P5, PV-immunoreactivity remained confined to layer V of the cortex, reminiscent of the expression profile manifested at the end of the second postnatal week in vivo. Also, the number of PV-immunoreactive cells in these cultures was significantly lower than in explants at the later stages. Our results indicate that the onset of PV-expression in the parietal cortex depends upon extrinsic cortical factors subsisting prior to P7. Once the production of this protein has been initiated, such influences are no longer required. PMID- 9753171 TI - Growth of pyramidal, but not non-pyramidal, dendrites in long-term organotypic explants of neonatal rat neocortex chronically exposed to neurotrophin-3. AB - The present study was undertaken to determine the effects of neurotrophin-3 (NT3) and spontaneous bioelectric activity (SBA) on dendritic elongation and branching in long-term isolated organotypic explants of rat neocortex. Viral vector directed expression of NT3 was used as an effective means to ensure a continuous, local production of the neurotrophic factor. Quantitative light microscopic measurement of dendritic branching patterns was carried out on Golgi-stained materials. Explants were exposed to an adenoviral vector encoding the genetic sequence for neurotrophin-3 (Ad-NT3), or to exogenous additions of the neuropeptide NT3. In order to test for activity-dependent growth effects under control and experimental conditions, explants were exposed to glutamatergic blockade using a cocktail of APV and DNQX. Both Ad-NT3 and NT3 peptide potently promoted apical and basal dendritic growth (elongation and branching) in pyramidal neurons. This growth was observed to be significant in layers II-IV and V. These growth effects were also not activity dependent, inasmuch as they were elicited from explants in which spontaneous bioelectric activity had been suppressed. Non-pyramidal neurons, throughout the neocortical slice, showed no significant dendritic responses to the prolonged presence of NT3. These findings show that pyramidal dendritic growth in long-term neocortical explants responds to at least one neurotrophic growth factor, NT3, and is independent of intrinsic bioelectric activity. The use of viral vectors in delivering a continuous high level of neurotrophic factor within developing neural tissues demonstrates its potential application to in vivo tissues during development, or in the stimulation of neuritogenesis and neuroregeneration following injuries. PMID- 9753172 TI - A rat G protein-coupled receptor selectively expressed in myelin-forming cells. AB - By screening an olfactory bulb cDNA library using dopamine receptor probes, we isolated the cDNA coding for the rat counterpart of an orphan receptor known as Edg-2, homologous to G protein-coupled receptors. In situ hybridization analysis showed that Edg-2 mRNA expression is restricted to myelinated structures, e.g. corpus callosum or peripheral nerves. A weaker expression in various peripheral organs was also detected in newborns. A 3.8-kb transcript was found at high levels in highly myelinated brain structures and sciatic nerve, and, at lower levels, in poorly myelinated peripheral organs, consistent with its occurrence in Schwann cells in the peripheral nervous system. One hundred percent of Edg-2 mRNA containing cells in the brain also expressed mRNA encoding myelin-basic-protein, a marker of oligodendrocytes. This restricted olygodendrocytes localization was confirmed by the absence of cellular colocalization of Edg-2 and glial fibrillary acidic protein, an astrocytic marker. During prenatal development, Edg-2 mRNA expression was high in the cortical neuroepithelium and meningeal layer at E16, extended later to other neuroepithelia, and disappeared shortly after birth. During brain postnatal development, Edg-2 mRNA expression in myelinated structures followed a caudo-rostral gradient, similar to that of myelination. Thus, Edg-2 is the first G protein-coupled receptor found to be selectively expressed in myelin-forming cells in the nervous system and its temporal expression pattern is consistent with a dual role (i) in neurogenesis, during embryonic development, and (ii) in myelination and myelin maintenance, during postnatal life. PMID- 9753173 TI - Lesion-induced transneuronal plasticity of the cholinergic innervation in the adult rat entorhinal cortex. AB - The present experiments were designed to determine the effect that lesions of the basal forebrain cholinergic system exert on cholinergic interneurons within the entorhinal cortex (EC) in the rat. Unilateral infusion of 192 IgG-saporin into the nucleus of the horizontal diagonal band of Broca (HDB) decreased the number of ipsilateral choline acetyltransferase immunoreactive (ChAT-ir) neurons by 54%. Two-four weeks after the lesion, the ipsilateral EC exhibited a moderate but significant loss of ChAT-ir fibres and interneurons. Adjacent sections revealed a parallel loss of vasoactive intestinal polypeptide (VIP) immunoreactivity. Cell counts in the cingulate cortex were unaffected, suggesting that this effect was indeed specific to the main target area for HDB neurons. Ibotenic acid lesions also induced a significant 36% decrease in the number of cholinergic neurons in the ipsilateral HDB, and disappearance of ChAT terminals in the EC, whereas the number of ChAT-ir neurons in the EC was unchanged. Since ibotenic acid affects all cells and not only cholinergic ones, our results suggest that the specific degeneration of cholinergic neurons in the HDB after 192 IgG-saporin treatment could be inducing transsynaptic effects on their targets. Injections of 192 IgG saporin directly into the EC also lesioned the cholinergic projection from the HDB, but had no effect on the intrinsic population. Eight weeks after immunolesion, the number of interneurons immunoreactive for ChAT and VIP in the EC had returned to normal values, and persisted for as long as 6 months after the lesion. By contrast, ChAT-ir neurons in the HDB were permanently lost. Our results suggest that the transient down-regulation of the cholinergic phenotype in entorhinal cortex interneurons could be a manifestation of activity-dependent plasticity, and that the loss of cholinergic innervation from the basal forebrain could be responsible for these effects through an imbalance of inputs. We hypothesize that the recovery of the phenotypic expression of entorhinal interneurons could be due to a recovery in their innervation, perhaps from sprouting axons in the same fields, belonging to surviving cholinergic neurons in the basal forebrain. PMID- 9753174 TI - Stimuli which entrain the circadian clock of the neonatal Syrian hamster in vivo regulate the phosphorylation of the transcription factor CREB in the suprachiasmatic nucleus in vitro. AB - Photic resetting of the adult mammalian circadian clock in vivo is associated with phosphorylation of the Ser133 residue of the calcium/cyclic AMP response element binding-protein (CREB) in the retinorecipient region of the suprachiasmatic nucleus (SCN). Western blotting and immunocytochemistry were used to investigate whether agonists known to reset the clock of neonatal hamsters in vivo are also able to influence the phosphorylation of CREB in the suprachiasmatic hypothalamus in vitro. Antisera raised against synthetic CREB peptide sequences were used to differentiate between total CREB and the Ser133 phosphorylated form of CREB (pCREB). Western blot analysis of proteins isolated from suprachiasmatic tissue of 1-day-old Syrian hamsters revealed bands at approximately 45 kDa corresponding to total CREB and pCREB. Treatment of the tissue with a mixture of glutamatergic agonists [N-methyl-D-aspartate (NMDA), amino-methyl proprionic acid (AMPA) and kainate, all at 1 microM], or native glutamate (1 microM) had no effect on the total CREB signal, but increased the pCREB signal, indicative of agonist-stimulated phosphorylation of CREB on Ser133. A similar effect was seen following treatment of the suprachiasmatic blocks with either dopamine (1 microM) or forskolin (1 microM). Simultaneous treatment with melatonin (1 microM) significantly attenuated stimulation by forskolin. The effect of the agonists on nuclear pCREB-immunoreactivity (-ir) was investigated in primary cultures which contained a mixture of cell types characteristic of the suprachiasmatic nuclei in vivo. Basal expression of nuclear total CREB-ir was high, whereas expression of pCREB-ir was low. Treatment with glutamate (1 microM) or dopamine (1 microM) had no effect on total CREB-ir, but increased pCREB-ir in approximately 50 and 30% of cells, respectively, whereas forskolin (1 microM) increased pCREB-ir in almost all cells (> 90%). The effects of all three agonists were rapid (< 15 min), and dose and time dependent. Melatonin reversed the effects of forskolin in mixed cultures, but not in pure astrocyte cultures. Dual immunocytochemistry (ICC) revealed that glutamate (1 microM) increased nuclear pCREB-ir in cells immunoreactive for microtubule-associated protein II (MAP II ir), but not other cells, indicating an effect predominantly on neurons. This occurred equally in gamma-amino butyric acid (GABA)-ir and non-GABA-ir neurons. Dopamine (1 microM) was more selective, increasing pCREB-ir only in GABA-ir neurons, whereas forskolin increased pCREB-ir in all cells. The specific stimulation of pCREB-ir in GABA-ir neurons by dopamine was reversed by melatonin, but melatonin had no effect on the increase in pCREB-ir induced in GABA-ir neurons by glutamate. These results demonstrate that agonists known to entrain the circadian clock in vivo modulate phosphorylation of CREB in GABA-ir neurons derived from the neonatal suprachiasmatic nuclei. PMID- 9753175 TI - Functional heterogeneity of periglomerular cells in the rat olfactory bulb. AB - The periglomerular cells of the rat olfactory bulb, a virtually unknown population of interneurons, have been studied applying the whole-cell patch-clamp technique to thin slices. A prominent result, obtained under current-clamp conditions, is that these cells appear to be functionally heterogeneous, and show distinct excitability profiles. Voltage-clamp analysis allows the identification of the ionic basis of these differences and suggests a division into at least two classes, based on the characteristics of the K+ conductances. The first group displays two K+ conductances (delayed rectifier, gKV, and fast transient, gA) of similar amplitude, and under current-clamp conditions shows the usual outward rectifying behaviour at depolarized potentials. The second group has a large gA, and a small or absent gKV. Consequently, following sustained depolarizations under current-clamp conditions leading to inactivation of gA, these neurons do not show any sign of outward rectification and behave as ohmic elements, as normally observed only at hyperpolarized potentials. The transition ion zinc (10 300 microM) affects gA but not gKV The inactivation process (steady-state curve and rate constant) is strongly altered by Zn2+, the activation process less so; open-channel conductance is not affected. The Zn2+ effect is unlikely to be due to surface charge screening or to a mechanism involving channel block. In view of the substantial presence of zinc ions in the olfactory nerve terminals, its actions on the A-current could be of some relevance for physiological function. PMID- 9753176 TI - Synaptic loss reflected by secretoneurin-like immunoreactivity in the human hippocampus in Alzheimer's disease. AB - Secretoneurin is a recently described peptide derived by endoproteolytic processing from secretogranin II, previously named chromogranin C. In this study, we have investigated the distribution of secretoneurin-like immunoreactivity in the human hippocampus in controls and in Alzheimer's disease patients, and compared the staining pattern to that of calretinin. Secretoneurin-like immunoreactivity is present throughout the hippocampal formation. At the border of the dentate molecular layer and the granule cell layer, a band of dense secretoneurin immunostaining appeared. In this part, as in the area of the CA2 sector, the high density of secretoneurin-immunoreactivity coincided with calretinin-like immunoreactivity. The mossy fibre system displayed a moderate density of secretoneurin-immunoreactivity. In the entorhinal cortex, a particularly high density of secretoneurin-immunoreactivity was observed. The density of secretoneurin-like immunoreactivity was significantly reduced in the innermost part of the molecular layer and in the outer molecular layer of the dentate gyrus in Alzheimer's disease. For calretinin-like immunoreactivity, a less pronounced decrease was found in the innermost part of the molecular layer. About 40-60% of neuritic plaques were secretoneurin-immunopositive. This study shows that secretoneurin is distinctly distributed in the human hippocampus and that significant changes of secretoneurin-like immunoreactivity occur in Alzheimer's disease, reflecting synaptic loss. PMID- 9753178 TI - Relations between cortical and thalamic cellular activities during absence seizures in rats. AB - In a rat model of generalized absence epilepsies (Genetic Absence Epilepsy Rats from Strasbourg, GAERS), multiunit activity was recorded simultaneously at different sites of the thalamocortical system under neurolept anaesthesia (fentanyl-droperidol). Under these conditions, bilaterally synchronized spike-and wave-discharges (SWDs) occurred spontaneously on the electroencephalogram (EEG) that were in principle identical to those reported earlier from unanaesthetized preparations. The generation of SWDs on the EEG was associated with spike concurrent, rhythmic burst-like activity in (mono-)synaptically connected regions of specific (somatosensory) thalamic regions and layers IVN of the somatosensory cortex, and the reticular thalamic nucleus. Precursor activity was typically recorded in cortical units, concomitant with 'embryonic' SW seizures on the EEG, before the paroxysm was evident on the gross EEG and in the thalamus. On average, SWD-correlated activity in layers IVN of the somatosensory cortex started significantly earlier than correlated burst-like firing in reticular and in ventrobasal thalamic neurons. Cellular peak firing in thalamus and cortex during bilaterally synchronized SWDs was related to the spike component on the gross EEG with the temporal rank order ventroposteromedial > ventrolateral > or = ventroposterolateral thalamic > > rostral reticular thalamic nuclei > or = cortex (layers IVN) = caudal reticular thalamic nucleus. A spike-related depression and wave-related increase in firing was recorded in anteroventral ventrolateral thalamic areas, presumably reflecting their peculiar anatomical arrangement within the thalamus. These results from an in vivo preparation with intact synaptic connections that spontaneously produces SWDs indicate that SWDs spread within the thalamocortical network, involving short and long delays. The order of concurrent rhythmic firing observed in thalamocortical circuits during SW seizures are supportive of the hypothesis that the processes of rhythmogenesis recruit local thalamic networks, while cortical mechanisms appear to synchronize rhythmic activities on a larger spatiotemporal scale, thereby providing an important contribution to the generalization of epileptiform activity and expression of SWDs on the EEG. PMID- 9753177 TI - The cortical somatotopic map and phantom phenomena in subjects with congenital limb atrophy and traumatic amputees with phantom limb pain. AB - The extent of the cortical somatotopic map and its relationship to phantom phenomena was tested in five subjects with congenital absence of an upper limb, four traumatic amputees with phantom limb pain and five healthy controls. Cortical maps of the first and fifth digit of the intact hand, the lower lip and the first toe (bilaterally) were obtained using neuroelectric source imaging. The subjects with congenital upper limb atrophy showed symmetric positions of the left and right side of the lower lip and the first toe, whereas the traumatic amputees with pain showed a significant shift (about 2.4 cm) of the cortical representation of the lower lip towards the hand region contralateral to the amputation side but no shift for the toe representation. In healthy controls, no significant hemispheric differences between the cortical representation of the digits, lower lip or first toe were found. Phantom phenomena were absent in the congenital but extensive in the traumatic amputees. These data confirm the assumption that congenital absence of a limb does not lead to cortical reorganization or phantom limbs whereas traumatic amputations that are accompanied by phantom limb pain show shifts of the cortical areas adjacent to the amputation zone towards the representation of the deafferented body part. PMID- 9753179 TI - The relation between dopamine oxidation currents in the nucleus accumbens and conditioned increases in motor activity in rats following repeated administration of d-amphetamine or cocaine. AB - Chronoamperometric recording techniques were used to monitor extracellular dopamine efflux in the nucleus accumbens associated with unconditioned and conditioned increases in motor activity in rats, following the intravenous administration of either d-amphetamine (0.63 mg/kg) or cocaine (3 mg/kg), or the presentation of a conditioned stimulus paired repeatedly with one of these psychostimulants. Each drug was administered daily for 7 days, either in the home cage or an environment in which a compound stimulus (light offset, odour) was presented. Rats in control groups received saline instead of drug in the distinctive test environment. On day 7 of training, significant increases in unconditioned motor activity were observed in the 30 min session following infusions of either d-amphetamine or cocaine. Associated dopamine oxidation currents in the nucleus accumbens increased immediately following administration of either drug and remained significantly elevated above baseline during the entire 30 min recording period. On the test day, presentation of the conditioned stimulus with vehicle infusions, in the distinct environment, was accompanied by an increase in dopamine oxidation currents and a conditioned increase in motor activity, only in the groups in which these stimuli had been paired with d amphetamine or cocaine. Neither the magnitude or duration of the conditioned motor activity matched the corresponding change in extracellular dopamine efflux in the nucleus accumbens. Accordingly, it is argued that the increase in dopamine concentration serves as a neurochemical correlate of the unconditioned and conditioned stimuli. The change in motor activity constitutes the unconditioned and conditioned responses that are subserved by the neural systems activated by the initial rise in extracellular dopamine. PMID- 9753180 TI - Conditioned changes in dopamine oxidation currents in the nucleus accumbens of rats by stimuli paired with self-administration or yoked-administration of d amphetamine. AB - In vivo chronoamperometry was used to monitor changes in dopamine oxidation currents corresponding to dopamine efflux in the nucleus accumbens of rats after presentation of a conditioned light stimulus repeatedly paired with either yoked- or self-administered intravenous injections of the psychostimulant d-amphetamine. Daily conditioning trials began with a non-contingent drug injection, paired with a conditioned stimulus consisting of a 5 s flashing light and 30 s lights out, after which a house light was illuminated during the 3 h session, signalling drug availability. Each subsequent injection of d-amphetamine was paired with the conditioned stimulus. Electrochemical measures were taken on conditioning trials 4-7, and on each trial, intravenous d-amphetamine (0.25 mg/kg per injection) self administration produced a significant maximal increase in mean dopamine oxidation currents of approximately 8 nA above baseline. Dopamine oxidation currents in rats receiving yoked d-amphetamine were approximately 5 nA above baseline by the fourth day of drug administration and reached approximately 8 nA on the seventh and final day of drug administration. On day 9 the first presentation of the vehicle injection and conditioned stimulus, in combination with illumination of the house lights, induced an immediate increase in nucleus accumbens dopamine oxidation currents in all rats that had previously received d-amphetamine. Subsequent presentations of the conditioned stimulus at 30 min intervals induced further increases in extracellular dopamine oxidation currents in both drug treated groups. By the end of the 3 h session, both groups had similar maximal conditioned increases in dopamine oxidation currents of approximately 6 nA. These data are discussed with relation to the neurochemistry of drug craving. PMID- 9753181 TI - Spatiotemporal distribution of a late synchronized activity in olfactory pathways following stimulation of the olfactory bulb in rats. AB - The evoked potential recorded in the rat piriform cortex in response to electrical stimulation of the olfactory bulb is composed of an early component occasionally followed by a late component (60-70 ms). We previously showed that the late component occurrence was enhanced following an olfactory learning. In the present study carried out in naive rats, we investigated the precise conditions of induction of this late component, and its spatiotemporal distribution along the olfactory pathways. In the anaesthetized rat, a stimulating electrode was implanted in the olfactory bulb. Four recording electrodes were positioned, respectively, in the olfactory bulb, the anterior and posterior parts of the piriform cortex, and the entorhinal cortex. Simultaneous recording of signals evoked in the four sampled structures in response to stimulation of the olfactory bulb revealed that the late component was detected in anterior and posterior piriform cortex as well as in entorhinal cortex, but not in the olfactory bulb. The late component occurred reliably for a narrow range of low intensities of stimulation delivered at frequencies not exceeding 1 Hz. Comparison of late component amplitude and latency across the different recorded sites showed that this component appeared first and with the greatest amplitude in the posterior piriform cortex. In addition to showing a functional dissociation between anterior and posterior parts of the piriform cortex, these data suggest that the posterior piriform cortex could be the locus of generation of this late high amplitude synchronized activity, which would then propagate to the neighbouring regions. PMID- 9753182 TI - Optokinetic reflex in squirrel monkeys after long-term monocular deprivation. AB - Horizontal optokinetic nystagmus (OKN) as well as neuronal response properties in the nucleus of the optic tract and the dorsal terminal nucleus of the accessory optic system (NOT-DTN) were investigated in three monocularly deprived squirrel monkeys. In two monkeys occlusion of one eye was performed at birth (early) and in the third after 7 weeks (late). In adulthood, in early deprived monkeys monocular horizontal OKN tested through the non-deprived eye was symmetrical and in no way different from normal, i.e. stimulation in the temporonasal and nasotemporal direction elicited equal and robust responses. OKN through the early occluded eye, however, was grossly abnormal with low gain and great variability in the consistency of nasotemporal and temporonasal slow phase eye movements. When in the late deprived monkey the non-deprived eye was occluded a strong spontaneous nystagmus developed despite the deprived eye viewing a stationary pattern. The slow phases were directed from nasal to temporal for the deprived eye. When tested through the non-deprived eye all neuronal responses of the NOT DTN were normal. The deprived eye's influence on NOT-DTN neurons was extremely weak. No neuron with a moderate or even dominant input from the deprived eye was found after early deprivation. In the late deprived case the deficit was not as severe but still the non-deprived eye was clearly dominating the responses in all neurons tested. Velocity tuning of neurons tested through the non-deprived eye was normal and qualitatively corresponded well to slow phase eye velocity in response to equivalent retinal slip during OKN. Through the early deprived eye, however, velocity tuning was extremely poor. It was somewhat better through the late deprived eye. We suggest that the dramatic deterioration in the optokinetic reflex found after long-term monocular deprivation for the amblyopic eye is probably caused by the almost complete loss of retinal and cortical input driven by that eye to the NOT-DTN. These results are discussed in relation to our previous results in cats and reports in the literature for humans with occlusion amblyopia. PMID- 9753183 TI - Immortalized gonadotropin-releasing hormone neurons secrete gamma-aminobutyric acid-evidence for an autocrine regulation. AB - The immortalized hypothalamic neuronal cell lines GT1-1 and GT1-7 represent unique model systems to investigate the physiological control of gonadotropin releasing hormone (GnRH) secretion. Using immunofluorescence microscopy, key proteins of regulated exocytosis, e.g. synaptotagmin, synaptobrevin and SNAP-25 (synaptosomal associated protein of 25 kDa) were found in GT1 neurons. In addition, GT1 neurons contained synaptophysin, a marker protein for small synaptic vesicles (SSVs) which are responsible for the storage of neurotransmitters such as gamma-aminobutyric acid (GABA). Upon subcellular fractionation, a lighter vesicle population characterized by synaptophysin separated from a denser vesicle population containing GnRH. Both vesicle populations contained synaptobrevin and synaptotagmin. Besides GnRH, GT1 neurons expressed glutamic acid decarboxylase at the mRNA-level and synthesized GABA. More importantly, GT1 neurons took up and stored 3H-GABA. The stored GABA was released after stimulation with increasing K+ concentrations and by alpha latrotoxin. Reducing the extracellular Ca2+-concentration abolished stimulated secretion, indicating that GABA was released by regulated exocytosis. Hormone secretion from GT1 neurons is controlled by GABA via GABA(A) and GABA(B) receptors reflecting the situation in vivo. Our data provide the first evidence that GT1 neurons possess a second regulated secretory pathway sustained by SSVs storing and releasing GABA. The released GABA influences GnRH secretion by an auto- or paracrine loop. PMID- 9753184 TI - Brain Ac39/physophilin: cloning, coexpression and colocalization with synaptophysin. AB - Physophilin is an oligomeric protein that binds the synaptic vesicle protein synaptophysin constituting a complex that has been hypothesized to form the exocytotic fusion pore. Microsequencing of several physophilin peptides putatively identified this protein as the Ac39 subunit of the V-ATPase. Ac39 has recently been shown to be present in a synaptosomal complex which, in addition to synaptophysin, includes the bulk of synaptobrevin II, and subunits c and Ac115 of the V0 sector of the V-ATPase. We have cloned physophilin from mouse brain and found a differential region of 12 amino acids when compared with the previously reported sequence of Ac39 from bovine adrenal medulla. RT-PCR cloning from the bovine adrenal medulla demonstrates that sequencing errors occurred in the previous cloning study, and shows that the amino acid sequences of physophilin and Ac39 are completely identical. In situ hybridization in rat brain reveals a largely neuronal distribution of Ac39/physophilin mRNA which spatio-temporally correlates with those of subunit c and synaptophysin. Immunohistochemical analysis shows that Ac39/physophilin is mostly concentrated in the neuropil with a pattern identical to subunit A and very similar to synaptophysin. Double labelling immunofluorescence shows a complete colocalization of Ac39/physophilin with subunit A and a partial colocalization with synaptophysin in the neuropil. Our findings bring anatomical support for the in vivo occurrence of the synaptophysin-Ac39/physophilin interaction and further suggest a coordinated transcription of V-ATPase and synaptophysin genes. A putative role of Ac39/physophilin in the inactivation of the V-ATPase by disassembly of its V1 sector is also discussed. PMID- 9753185 TI - Scalp recorded direct current brain potentials during human sleep. AB - The direct current (DC) potential recorded from the scalp of awake humans has been considered a reflection of general changes in cortical excitability. This study examined DC potential shifts in humans during a night of continuous sleep. Standard polysomnographic recordings and skin temperature were measured simultaneously. Contrary to expectations, average DC potential level indicated higher negativity during nonrapid eye movement (NREM) sleep than REM sleep and wakefulness. Moreover, a dynamic regulation of the DC potential level was revealed in association with the NREM-REM sleep cycle comprising four successive phases: (i) a steep 'NREM-transition-negative shift' during the initial 10-15 min of the NREM sleep period; (ii) a more subtle 'NREM-positive slope' during the subsequent NREM sleep period; (iii) a steep 'REM-transition-positive shift' starting shortly prior to the REM sleep period, and (iv) a 'REM-negative slope', characterizing the remaining greater part of the REM sleep period. DC potential changes were only weakly related to changes in slow-wave activity (r2 < 0.18). The NREM-negative slope and REM-positive slope could reflect, respectively, gradually increasing and decreasing cortical excitability resulting from widespread changes in the depolarization of apical dendrites. In contrast, the NREM-transition-negative shift and the REM-transition-positive shift may reflect the progression and retrogression, respectively, of a long-lasting hyperpolarization in deeply lying neurons. PMID- 9753186 TI - A dopamine-mu1 opioid link in the rat ventral tegmentum shared by palatable food (Fonzies) and non-psychostimulant drugs of abuse. AB - The role of mu1 opioid receptors in the stimulation of dopamine transmission in the rat nucleus accumbens by an unusual palatable food (Fonzies) and non psychostimulant drugs of abuse was investigated by the use of naloxonazine, a pseudo-irreversible antagonist of mu1 opioid receptors. Feeding of Fonzies stimulated dopamine release in the medial prefrontal cortex and in the shell, but not in the core of the nucleus accumbens. Pretreatment with naloxonazine given systemically (15 mg/kg i.p. 20 h before) completely prevented the stimulation of dopamine release in the shell of the nucleus accumbens by Fonzies without affecting that in the prefrontal cortex. Systemic pretreatment with naloxonazine reduced or, depending on the dose, abolished, the stimulation of dopamine release in the nucleus accumbens shell by morphine, nicotine and ethanol, but did not affect that by haloperidol. Naloxonazine also prevented the stimulatory effects of Fonzies, nicotine and morphine on nucleus accumbens dopamine transmission when infused bilaterally in the ventral tegmental area. The results indicate that mu1 opioid receptors in the ventral tegmentum play a major role in the stimulant effects of food and drugs of abuse on mesolimbic dopamine transmission. PMID- 9753187 TI - Interleukin-1beta and the interleukin-1 receptor antagonist act in the striatum to modify excitotoxic brain damage in the rat. AB - The cytokine interleukin-1 (IL-1) has been implicated in ischaemic, traumatic and excitotoxic brain damage. The results presented here reveal novel actions of IL-1 in the striatum which markedly exacerbate cortical neuronal damage elicited by local excitotoxins in the striatum or cortex. Intrastriatal infusion of IL-1 receptor antagonist, IL-1ra, markedly inhibited striatal neuronal damage caused by N-methyl-D-aspartate (NMDA) or alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionate (AMPA) receptor activation in the rat. In contrast, intracortical infusion of IL-1ra failed to inhibit NMDA or AMPA receptor-induced damage in the cortex. Intrastriatal co-infusion of IL-1 with the NMDA or AMPA receptor agonist did not affect local striatal damage induced by activation of either glutamate receptor subtype, but caused extensive cortical damage when administered into the striatum with AMPA. This secondary damage was significantly reduced by pretreatment with the NMDA receptor antagonist (MK-801), which did not affect local (striatal) damage caused by AMPA. Infusion of IL-1beta into the striatum (but not the cortex) markedly enhanced cortical damage caused by infusion of an NMDA or AMPA receptor agonist into the cortex. These data reveal selective actions of IL-1 and IL-1ra in the striatum, which influence cortical neuronal loss and suggest that IL-1 selectively enhances damage caused by AMPA receptor activation. PMID- 9753188 TI - Opioid withdrawal activates MAP kinase in locus coeruleus neurons in morphine dependent rats in vivo. AB - Opioid dependence is widely believed to result from neuroadaptations in specific brain regions. However, the precise molecular mechanisms underlying these adaptations are not yet clear. Our aim was to explore the role of mitogen activated protein kinase (MAPK) in mu opioid receptor signalling in vivo. Using anti-phospho MAPK antibodies, activated MAPK was detected in cortical neurons (layers II/III), median eminence, amygdaloid and hypothalamic nuclei in untreated animals. Dense nuclear and cytoplasmic staining was observed resulting in full visualization of processes in these cells. Chronic, but not acute, administration of morphine greatly diminished this staining pattern while mu opioid receptor levels and levels of MAP kinase as detected with a phosphorylation state independent antibody were unchanged. When opioid withdrawal was precipitated with naloxone a dramatic increase in MAP kinase phosphorylation was observed in somata and fibres of locus coeruleus, solitary tract and hypothalamic neurons. Thus, the differential activation state of MAPK could have important implications for understanding the mechanisms underlying opioid tolerance and dependence. PMID- 9753190 TI - No-go dominant brain activity in human inferior prefrontal cortex revealed by functional magnetic resonance imaging. AB - We investigated the response inhibition function of the prefrontal cortex associated with the go/no-go task using functional magnetic resonance imaging in five human subjects. The go/no-go task consisted of go and no-go trials given randomly with roughly equal probability. In go trials a green square was presented and the subjects had to respond by promptly pushing a button using their right or left thumbs, but in no-go trials a red square was presented and subjects were instructed not to respond. When brain activity in no-go trials is dominant over that in go trials in areas in the prefrontal cortex, this no-go dominant brain activity would reflect the neural processes for inhibiting inherent response tendency. We used a new strategy of image data analysis by which transient brain activity in go or no-go trials can be analysed separately, and looked for the prefrontal areas in which the brain activity in no-go trials is dominant over that in go trials. We found the no-go dominant foci in the posterior part of the right inferior frontal sulcus reproducibly among the subjects. This was true whether the right or left hand was used. These results suggest that this region in the prefrontal cortex is related to the neural mechanisms underlying the response inhibition function. PMID- 9753189 TI - Cloning of a new gap junction gene (Cx36) highly expressed in mammalian brain neurons. AB - The connexins are the protein subunits of the gap junction intercellular channels. In the present study a new rat connexin was cloned by degenerate reverse transcription-polymerase chain reaction and its gene isolated from a mouse genomic library. The nucleotide sequence encodes a protein of 321 amino acids (called Cx36) with highly significant homology to the members of the connexin family. In situ hybridization analysis of rat brain and retina showed the strongest expression in neurons of the inferior olive, the olfactory bulb, the CA3/CA4 hippocampal subfields and several brain-stem nuclei. An intense expression was also found in the pineal gland and in the retinal ganglion cell and inner nuclear layers. Experiments with neurotoxins, locally injected in the hippocampus or specifically acting on inferior olivary neurons, confirmed the neuronal localization of Cx36. It is the first connexin to be expressed predominantly in mammalian neurons and its identification paves the way for a molecular approach in the study of the role played by gap junctions in the physiology and the pathology of the mammalian brain. PMID- 9753191 TI - Role of the CNS microvascular pericyte in the blood-brain barrier. AB - Pericytes are a very important cellular constituent of the blood-brain barrier. They play a regulatory role in brain angiogenesis, endothelial cell tight junction formation, blood-brain barrier differentiation, as well as contribute to the microvascular vasodynamic capacity and structural stability. Central nervous system pericytes express macrophage functions and are actively involved in the neuroimmune network operating at the blood-brain barrier. They exhibit unique functional characteristics critical for the pathogenesis of a number of cerebrovascular, neurodegenerative, and neuroimmune diseases. PMID- 9753192 TI - Interleukin-1 and nociception in the rat. AB - It has recently become accepted that several cytokines may affect peripheral and central nervous system functions. Consistently with these findings, accumulating evidence points toward an important role for interleukin- in the modulation of nociceptive information. Here we review the observations collected after the administration of this cytokine by intracerebroventricular, intrathecal or peripheral route in rats. Taken together, these data suggest that IL-1 can differently affect pain responsivity depending on the dose and the site of action, and clearly demonstrate that this immune factor is deeply involved in the modulation of neuronal functions. PMID- 9753193 TI - Opposing mitogenic regulation by PACAP in sympathetic and cerebral cortical precursors correlates with differential expression of PACAP receptor (PAC1-R) isoforms. AB - Neurogenesis in the peripheral and central nervous systems proceeds in region specific fashion, although underlying mechanisms remain undefined. Emerging evidence indicates that the neuropeptide PACAP and its G-protein-coupled receptor are expressed widely in the embryonic brain, suggesting that the ligand/receptor system plays a role in development. We found previously that PAC1-R activation elicited opposing mitogenic effects in neurogenetic cultures, stimulating peripheral sympathetic neuroblasts while inhibiting cerebral cortical precursors. We have now defined the expression of PAC1-R mRNA isoforms and activation of second-messenger pathways in these model populations. Sympathetic neuroblasts express the "hop" receptor isoform, through which PACAP elicits increased levels of cAMP and activation of the PI signaling pathway. In contrast, cerebral cortical precursors express primarily the "short" (non-insert) receptor isoform and exhibit increased cAMP levels alone following PACAP treatment. Thus, opposing mitogenic regulation in sympathetic and cortical precursors correlates with differential receptor isoform expression and distinct second-messenger signaling. In addition to receptor, PACAP ligand mRNA was expressed by both populations, suggesting that the peptide is produced and acts locally to regulate precursor proliferation. These observations indicate that the PACAP ligand/receptor system is expressed in both the peripheral and central nervous system during development. More generally, these studies suggest that widely expressed extracellular factors mediate region-specific neurogenesis by activating lineage restricted receptor isoforms and intracellular pathways. PMID- 9753194 TI - Muscle could be the therapeutic target in SMA treatment. AB - A nerve-muscle coculture model (human muscle cells innervated by embryonic rat spinal cord) was used to explore the pathogenesis of spinal muscular atrophy (SMA). Previous studies showed that myofibers from donors with SMA type I or SMA type II (but not SMA type III) undergo a characteristic degeneration 1-3 weeks after innervation (Braun et al. [1995] Lancet 345:694-695). To determine which cells are involved in degeneration, we cloned satellite cells and fibroblasts derived from muscle biopsies of normal (healthy) donors and donors with SMA. We show that fibroblasts are required for successful innervation, that fibroblasts from normal and SMA donors contribute equally well to the establishment of cocultures, and that only SMA satellite cells are responsible for the degeneration of innervated cocultures. We succeeded in preventing the degeneration of cloned satellite cells from SMA donors by adding 50% cloned satellite cells from normal donors to the culture to make heteromyotubes. In mixed cocultures, after innervation, we did not observe degeneration. This result suggests that survival of the cocultures depends on a message derived from the muscle cells. Consequently, we propose that therapeutic approaches for SMA that could repair (or compensate for) the genetic defect in muscle cells (which are otherwise much more accessible for gene therapy than neurons) might prevent motoneuron degeneration. The role of muscle cells in the establishment and the degeneration of neuromuscular junctions deserves further attention and investigation. PMID- 9753195 TI - Differential turnover of syntaxin and SNAP-25 during synaptogenesis in cultured cerebellar granule neurons. AB - In order to investigate the molecular mechanism underlying synaptogenesis, we examined the dynamics and stability of syntaxin 1A and SNAP-25 in cultured cerebellar granule cells. In neurons cultured for less than 5 days in vitro (DIV), syntaxin was highly expressed with a half-life of >48 hours. SNAP-25 was also expressed at 5 DIV, but at a lower level and with a much shorter half-life of 16 hours. As the neurons matured and established synpatic connections, the expression of both proteins increased steadily, with the more rapid increase between 5 DIV and 8 DIV associated with SNAP-25. The half-life of syntaxin was slightly increased in the mature neurons. SNAP-25, however, showed an increased half-life of about 35 hours. These results suggested that the dynamics and stability of the t-SNAREs are differentially modulated during synaptogenesis, which may be important in establishing and maintaining synaptic connections. PMID- 9753196 TI - Recombinant plasma-type platelet-activating factor acetylhydrolase attenuates NMDA-induced hippocampal neuronal apoptosis. AB - The bioactive lipid platelet-activating factor (PAF) accumulates in brain during injury, seizures and ischemia and may, in addition, be significant in AIDS dementia and in other neurodegenerative diseases. We have used plasma-type recombinant PAF acetylhydrolase (rPAF-AH) to test the hypothesis that PAF accumulation is involved in early events leading to neuronal apoptosis during excitotoxic neuronal injury. Neuronal cultures were labeled with FITC-12-dUTP (TUNEL technique) and propidium iodide, digitized using fluorescence microscopy and a chilled 3CCD color camera, and analyzed with 2D graphics analysis software. N-methyl-D-aspartate (NMDA) (50 microM, 2 hr) induced a 2.5-fold increase in apoptosis of hippocampal neurons compared with controls when analyzed 24 hr after NMDA treatment. Hippocampal neurons receiving rPAF-AH (20 microg/ml) before, during, and after NMDA treatment demonstrated a concentration-dependent neuroprotective effect which resulted in 47% and 30% neuroprotection against 50 and 100 microM NMDA, respectively. The noncompetitive NMDA receptor antagonist MK 801(300 nM) completely inhibited apoptosis caused by NMDA. The neuroprotective effect of rPAF-AH against NMDA-induced apoptosis was confirmed using as additional criteria, histone release, electron microscopy, and DNA laddering. Neuroprotection elicited by rPAF-AH demonstrates that PAF is an injury mediator in NMDA-induced neuronal apoptosis and that the recombinant protein is potentially useful as a therapeutic approach. PMID- 9753197 TI - Lysophosphatidic acid and apoptosis of nerve growth factor-differentiated PC12 cells. AB - The lipid biomediator lysophosphatidic acid (LPA) elicits a unique response in hippocampal neurons, LPA induces neuronal apoptosis. This study explores the effects of LPA on cells with neuronal properties, nerve growth factor differentiated PC6 cells, a clone of PC12 cells. LPA induced apoptosis in these cells as assessed by chromatin condensation, terminal dUTP nick end-labeling of DNA, protection against these nuclear alterations by a general caspase inhibitor and the lack of release of lactic dehydrogenase. LPA caused oxidative stress, namely a decreased reduction of MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide. This oxidative stress appears to be of functional significance, since cells were protected by pretreatment with the antioxidant propyl gallate and by stable transfection with cDNA encoding the antioxidant enzyme, manganese superoxide dismutase. Mitochondrial and nitric oxide participation in LPA-induced apoptosis are suggested by the protection afforded by pretreatment with either cyclosporin A, an inhibitor of mitochondrial permeability transition, or nitric oxide synthase inhibitors. The nitric oxide synthase inhibitor findings are novel, since to our knowledge, LPA has not heretofore been associated with an increase in nitric oxide. In addition, as observed for many neurotoxic agents, insulin-like growth factor I protected against LPA-induced apoptosis of PC6 cells. PMID- 9753198 TI - Abnormal astrocyte development and neuronal death in mice lacking the epidermal growth factor receptor. AB - Stimulation of the epidermal growth factor receptor (EGF-R) produces numerous effects on central nervous system (CNS) cells in vitro including neuronal survival and differentiation, astrocyte proliferation and the proliferation of multipotent progenitors. However, the in vivo role of EGF-R is less well understood. In the present study, we demonstrate that EGF-R null mice generated on a 129Sv/J Swiss Black background undergo focal but massive degeneration the olfactory bulb, piriform cortex, neocortex, and thalamus between postnatal days 5 and 8 which is due, at least in part, to apoptosis. Some of the neuronal populations that degenerate do not normally express EGF-R, indicating an indirect mechanism of neuronal death. There were also delays in GFAP expression within the glia limitans and within structures outside the germinal zones in early postnatal ages. At or just prior to the onset of the degeneration, however, there was an increase in GFAP expression in these areas. The brains of EGF-R (-/-) animals were smaller but cytoarchitecturally normal at birth and neuronal populations appeared to be intact, including striatal GABAergic and midbrain dopaminergic neurons which have previously been shown to express EGF-R. Multipotent progenitors and astrocytes derived from EGF-R (-/-) mice were capable of proliferating in response to FGF-2. These data demonstrate that EGF-R expression is critical for the maintenance of large portions of the postnatal mouse forebrain as well as the normal development of astrocytes. PMID- 9753199 TI - Traumatic brain injury causes delayed motor and cognitive impairment in a mutant mouse strain known to exhibit delayed Wallerian degeneration. AB - Delayed Wallerian degeneration after neuronal injury is a feature of the C57BL/Wld(s) mouse mutant. In the present study, we examined the effect of unilateral controlled cortical impact (CCI) on motor and cognitive performance in C57BL/6 and C57BL/Wld(s) mice. Performance on a beam-walking task was impaired in both injured groups over the first 3 weeks; however, between 28 and 35 days post injury, C57BL/6 mice continued to improve whereas C57BL/Wld(s) mice showed increased footfaults. In a spatial learning task, C57BL/Wld(s) animals performed consistently better than C57BL/6 mice when tested 7-10 days and 14-17 days following CCI. C57BL/Wld(s) mice also demonstrated improved working memory performance as compared with C57BL/6 mice when trained on days 21-22 after injury; this effect was lost on days 23 and 24, and was not evident in other animals tested in the same task at 28-31 days following injury. These results indicate a marked delay in motor and cognitive impairment following CCI in C57BL/Wld(s) mice compared with injured C57BL/6 controls. This is consistent with previous work showing delayed temporal evolution of neuronal degeneration in C57BL/Wld(s) mice and suggests CCI may be a suitable model for examining the functional consequences of traumatic brain injury (TBI) in genetically altered mice. PMID- 9753200 TI - Calcium-induced activation of the mitochondrial permeability transition in hippocampal neurons. AB - The mitochondrial permeability transition (mPT) has been implicated in both excitotoxic and apoptotic neuronal cell death, despite the fact that it has not been previously identified in neurons. To study the mPT in hippocampal neurons, cultures were loaded with the mitochondrial dye JC-1 and observed with confocal and conventional microscopy. After pretreatment with 4Br-A23187 and subsequent calcium addition, the initially rodlike mitochondria increased in diameter until mitochondria became rounded in appearance. Morphological changes reversed when calcium was removed by EGTA. When neurons were loaded with both fura-2-AM and rhodamine 123, calcium loading produced an increase in cytosolic calcium, mitochondrial depolarization, and similar alterations in mitochondrial morphology. Smaller calcium challenges produced calcium cycling, delaying morphological changes until after secondary depolarization and calcium release to the cytosol. In neurons exposed to glutamate, confocal observation of JC-1 fluorescence revealed comparable changes in mitochondrial morphology that were prevented when barium was substituted for calcium, or following pretreatment with the mPT inhibitor, cyclosporin A. These experiments establish conditions in which the mPT could be observed in situ in neurons in response to calcium loading. In addition, the timing of changes suggested that induction of the permeability transition in situ represents a sequence of multiple events that may reflect the multiple open conformations of the mPT pore. PMID- 9753201 TI - 1,25-Dihydroxyvitamin D3 regulates the expression of VDR and NGF gene in Schwann cells in vitro. AB - The vitamin D receptor (VDR) is a nuclear receptor that mediates the effect of the active metabolite of vitamin D3, the 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3). To investigate the potential role of this hormone in the peripheral nervous system, we have studied the VDR expression in Schwann cells. The VDR mRNA was detected by Northern blot analysis in rat primary cultures of Schwann cells, and its levels were strongly increased in the presence of 1,25-(OH)2D3. Using the mouse Schwann cell line, MSC80, we showed that concentrations as low as 10(-10) M of hormone stimulated the expression of the VDR gene and strongly increased the amounts of activated VDR, capable of binding to the specific vitamin D responsive element (VDRE). We also found that 1,25-(OH)2D3 stimulated the expression of the nerve growth factor gene in MSC80. These data suggest a role for the hormone in the peripheral nervous system, possibly as a mediator active in trauma. PMID- 9753202 TI - TNF-alpha and TGF-beta act synergistically to kill Schwann cells. AB - Interactions between cytokines and Schwann cells (SC) are important in development, repair, and disorders of the peripheral nervous system (PNS). Tumor necrosis factor-alpha (TNF-alpha) and transforming growth factor-beta (TGF-beta) are two prominent cytokines which may be involved in these processes and their gene products are upregulated in some experimental neuropathies. This study focuses on the in vitro effects of these cytokines, both singly and in combination, on cultured SC. Expression of both Type I and Type II TNF-alpha receptors was demonstrated on the SC surface by immunocytochemistry. Treatment of SC with a combination of TNF-alpha plus TGF-beta causes significant detachment and cell death while treatment with each cytokine alone is not significantly cytotoxic. When compared with control cultures, SC treated with the combination of cytokines exhibit an increase in the number of cells with condensed nuclei and evidence of DNA fragmentation, characteristics consistent with cells undergoing programmed cell death. Thus, TNF-alpha plus TGF-beta induce SC loss of adhesion which is predominantly due to cell death. Apoptotic mechanisms are likely to contribute to some extent to this cell death. These findings provide in vitro evidence to support the hypothesis that cytokines can directly damage SC in PNS disorders. PMID- 9753203 TI - Innovative pharmacologic approaches to cardiopulmonary resuscitation. AB - The survival rate of patients undergoing cardiopulmonary resuscitation (CPR) is 5% to 15%. New treatment approaches under investigation for CPR include the use of vasopressin as a vasopressor, amiodarone for the treatment of ventricular tachyarrhythmia, and adenosine antagonists (i.e., theophylline) for bradyasystolic rhythms. More innovative approaches include the use of thyroid hormone and endothelin. PMID- 9753204 TI - Problems and opportunities in regulation of benzodiazepines. AB - A commentary on current national and international regulation of benzodiazepine like drugs is presented. The policies and decision making associated with these regulations also are discussed, both in terms of historical mistakes and current problems. Finally, a proposal is made to improve the methods by which regulations are put into effect. PMID- 9753205 TI - A pharmacoeconomic model to aid in the allocation of ambulatory clinical pharmacy services. AB - Drugs of choice in secondary prevention strategies reduce complication rates of certain diseases. Unfortunately, these strongly indicated drugs remain underused. A model was developed to predict the cost-effectiveness of clinical pharmacy services assumed to improve use of drugs of choice to unity in hypothetical cohorts of three diseases that commonly accompany hypertension and in which clear drugs of choice exist. Use of angiotensin-converting enzyme (ACE) inhibitors in patients with diabetes who have proteinuria, use of beta blockers after myocardial infarction, and use of ACE inhibitors in patients with asymptomatic left ventricular dysfunction were analyzed. Clinical pharmacy services could be cost-saving in all three diseases in this model if use of the drug of choice in standard practice did not exceed 0.899 in patients with diabetes who have proteinuria, 0.512 in patients after infarct, and 0.804 in patients with asymptomatic left ventricular dysfunction. This model may help decision makers by accessing local patient demographics and prescribing habits before any resource allocation. PMID- 9753206 TI - The pharmacokinetics of intravenous testosterone in elderly men with coronary artery disease. AB - Intracoronary testosterone injections have recently been shown to possess coronary vasodilating effects. The same may be true for intravenous testosterone, but the pharmacokinetic and hemodynamic aspects need exploration before pharmacologic studies can begin. This trial determined the pharmacokinetic and hemodynamic properties of 300 microg of testosterone given intravenously. Degree of testosterone aromatization to 17-beta estradiol after exogenous administration and overall patient tolerability also were evaluated. Eleven elderly men with coronary artery disease participated in the study and were given 300 microg of testosterone intravenously over 10 minutes. Serum blood concentrations of testosterone and 17-beta estradiol were measured at baseline and then periodically. Testosterone serum concentrations were stripped and fit to a two compartment model for all patients. The volume of distribution (Vdarea) was 80.36 +/- 24.51 L, and the elimination half-life was 55.93 +/- 23.06 minutes. No hemodynamic differences or side effects were noted. The serum concentrations of 17-beta estradiol were significantly increased from baseline beginning 5 minutes after infusion to the end of the study (180 minutes after infusion). PMID- 9753207 TI - Pharmacokinetics of oral and intravenous dolasetron mesylate in patients with renal impairment. AB - In an open-label, randomized, two-way complete crossover study, the influence of renal impairment on the pharmacokinetics of dolasetron and its primary active metabolite, hydrodolasetron, were evaluated. Patients with renal impairment were stratified into three groups of 12 based on their 24-hour creatinine clearance (Cl(cr)): group 1, mild impairment (Cl(cr) between 41 and 80 mL/min); group 2, moderate impairment (Cl(cr) between 11 and 40 mL/min); and group 3, endstage renal impairment (Cl(cr) < or = 10 mL/min). Twenty-four healthy volunteers from a previous study served as the control group. Each participant received a single intravenous or oral 200-mg dose of dolasetron mesylate on separate occasions. Serial blood samples were collected up to 60 hours after dose for determination of dolasetron and hydrodolasetron, and urine samples were collected in intervals up to 72 hours for determination of dolasetron, hydrodolasetron, and the 5' and 6'-hydroxy metabolites of hydrodolasetron. Because plasma concentrations were low and sporadic, pharmacokinetic parameters of dolasetron were not calculated after oral administration. Although some significant differences in area under the concentration-time curve (AUC0-infinity), volume of distribution (Vd), systemic clearance (Cl), and elimination half-life (t1/2) of the parent drug were observed between control subjects and patients with renal impairment, there were no systematic findings related to degree of renal dysfunction. The elimination pathways of hydrodolasetron include both hepatic metabolism and renal excretion. Consistent increases in mean Cmax, AUC0-infinity, and t1/2 and decreases in renal and total apparent clearance of hydrodolasetron were seen with diminishing renal function after intravenous administration of dolasetron mesylate. No consistent changes were found after oral administration. Urinary excretion of hydrodolasetron and its metabolites decreased with decreasing renal function, but the profile of metabolites remained constant. Dolasetron was well tolerated in all three groups of patients. Based on these findings, no dosage adjustment for dolasetron is recommended in patients with renal impairment. PMID- 9753208 TI - Comparison of two cyclosporine formulations in healthy volunteers: bioequivalence of the new Sang-35 formulation and Neoral. AB - This study was conducted to establish bioequivalence between a newly developed oral cyclosporine formulation, Sang-35 (SangStat Medical Corp., Menlo Park, CA), and the microemulsion formulation Neoral (Novartis Pharmaceuticals, East Hanover, NJ). In a randomized, open-label, two-way crossover study, 36 fasted, healthy male volunteers received a single 500-mg cyclosporine dose formulated either as Sang-35 or Neoral. Mean are under the concentration-time curve to infinity (AUC0 infinity) for Sang-35 was 13,900 microg x hr/L compared with 14,000 microg x hr/L for Neoral, with a 90% confidence interval (CI) of 96% to 103% for the geometric mean ratio of the two formulations. Mean maximum concentration (Cmax) was 1,690 microg/L for Sang-35 and 1,700 microg/L for Neoral, with a 90% CI of 96% to 103%. Geometric mean ratios for both AUC0-infinity and Cmax were within the acceptance criteria for bioequivalence (80-125%). Additional studies showed no differences between Sang-35 and Neoral after high-fat meals (n = 19), in female volunteers (n = 25) and in black volunteers (n = 7). It is concluded that single doses of the oral cyclosporine formulations Sang-35 and Neoral are bioequivalent in healthy fasted subjects, after high-fat meals, in women, and in blacks. PMID- 9753209 TI - Effect of troglitazone on urinary excretion of 6beta-hydroxycortisol. AB - Coadministration of troglitazone reduces the plasma concentrations of terfenadine and ethinyl estradiol. Because these drugs are metabolized at least in part by cytochrome P450 3A (CYP3A), it is possible that troglitazone induces CYP3A activity, thereby reducing plasma concentrations of these agents. Known inducers of CYP3A, such as rifampin, phenytoin, carbamazepine, and phenobarbital, increase the urinary excretion of 6beta-hydroxycortisol and the ratio of 6beta hydroxycortisol to cortisol. This evaluation examined the effect of troglitazone on urinary excretion of 6beta-hydroxycortisol and the ratio of 6beta hydroxycortisol to cortisol as a marker for CYP3A induction. Urine samples were collected from 11 subjects who completed a study evaluating the effect of multiple-dose administration of troglitazone 400 mg once daily (days 11-20) on the steady-state pharmacokinetics of digoxin (0.25 mg daily on days 1-20). A single urine sample was collected at baseline on day 1, and 24-hour urine samples were collected on days 10 and 20. Mean +/- standard deviation 24-hour excretion rate of cortisol was unchanged, whereas that of 6beta-hydroxycortisol increased during troglitazone administration. The ratio of 24-hour urinary 6beta hydroxycortisol to cortisol excretion increased from 7.4 +/- 2.7 on day 10 to 16.1 +/- 6.2 on day 20. The ratio observed on day 10 was similar to that obtained at baseline. These results are consistent with the hypothesis that troglitazone induces CYP3A activity. PMID- 9753210 TI - Coadministration of acetaminophen and troglitazone: pharmacokinetics and safety. AB - Troglitazone, a PPAR-gamma agonist, enhances the actions of insulin on muscle and liver. It is metabolized predominantly in the liver to a sulfate conjugate and a quinone metabolite. Acetaminophen also undergoes metabolism by conjugation. This three-way crossover study in 12 healthy male volunteers was conducted to investigate the effects of acetaminophen on the metabolism of troglitazone and vice versa. No statistically or clinically relevant differences in area under the concentration-time curve extrapolated to infinity (AUCinfinity) were observed for troglitazone, its quinone metabolite, or acetaminophen. A statistically significant decrease in troglitazone sulfate conjugate during administration with acetaminophen was not clinically relevant. No statistically or clinically relevant differences were observed in maximum concentration (Cmax), time to Cmax (tmax), or elimination half-life of troglitazone, its two main metabolites, and acetaminophen or in acetaminophen urinary sulfate excretion, although there was a slight decrease in acetaminophen glucuronide excretion during administration with troglitazone. Adverse events were minor and similar between treatments. These findings suggest that troglitazone and acetaminophen can be coadministered without adverse clinical consequences. PMID- 9753211 TI - Influence of ondansetron on thiopental hypnotic requirements. AB - Ondansetron, a selective 5-HT3 antagonist has been proved to be an effective antiemetic agent for prophylaxis of nausea and vomiting after surgery. This study was conducted to determine whether ondansetron changes thiopental requirements for induction of anesthesia in patients unpremedicated and premedicated with diazepam. One hundred sixty eight adult female patients classified as American Society of Anesthesiologists (ASA) physical status I (normal healthy patient) or II (patient with mild systemic disease) participated in this prospective, double blinded, randomized study. Patients were assigned to receive either 0.07 mg/Kg diazepam orally or no premedication. They then received saline and ondansetron 0.1 mg/Kg or 0.2 mg/Kg intravenously 5 minutes before thiopental induction. Thiopental was administered at a rate of 25 mg/min until the patient lost the ability to open eyes on command. Thiopental requirements were not significantly different among groups. The results indicate that ondansetron in clinically used doses does not influence the hypnotic requirements of thiopental. PMID- 9753212 TI - Comparison of azithromycin and clarithromycin in their interactions with rifabutin in healthy volunteers. AB - A 14-day, randomized, open, phase I clinical trial was designed to examine possible pharmacokinetic interactions between rifabutin and two other antibiotics, azithromycin and clarithromycin, used in the treatment of Mycobacterium avium complex infections. Thirty healthy male and female volunteers were divided into five groups of six participants each: 18 received 300 mg/day of rifabutin, 12 in combination with therapeutic doses of either azithromycin or clarithromycin; the remaining 12 received azithromycin or clarithromycin alone. On day 10 the study was terminated because of adverse events, including severe neutropenia. Fourteen participants who received rifabutin developed neutropenia, including all 12 participants who received azithromycin or clarithromycin concomitantly. Analyses of serum revealed no apparent pharmacokinetic interaction between azithromycin and rifabutin. However, the mean concentrations of rifabutin and 25-O-desacetyl-rifabutin (an active metabolite) in participants who received clarithromycin and rifabutin concomitantly were more than 400% and 3,700%, respectively, of concentrations in those who received rifabutin alone. Physicians should be aware that recommended prophylactic doses of rifabutin may be associated with severe neutropenia within 2 weeks after initiation of therapy, and all patients receiving rifabutin, especially with clarithromycin, should be monitored carefully for neutropenia. PMID- 9753213 TI - Mechanisms of excitatory amino acid release in contused brain tissue: effects of hypothermia and in situ administration of Co2+ on extracellular levels of glutamate. AB - In order to elucidate the mechanisms of release of EAAs and their excitotoxicity in cerebral contusion, cortical contusion was produced in the rat parietal cortex, and the changes in extracellular levels of EAAs in the central and peripheral areas of contusion were investigated using microdialysis. The cortical contusion induced a rapid increase in dialysate concentration of glutamate ([Glu]d) from a baseline level of 4.6+/-2.8 microM to a maximum level of 36.3+/ 12.8 microM. This elevation of glutamate was significantly attenuated by mild hypothermia (32 degrees C for 90 min, comprising 20 min before and 70 min after the injury induction) in the peripheral area of contusion (p < 0.01) but not in the central area. Histological evaluations revealed that the hypothermia reduced the necrosis volume of contusion to 38.3% of that in the normothermic control (p < 0.01). In situ administration of Co2+, an inhibitor of Co2+-dependent exocytotic release of EAAs from the nerve terminals, via the microdialysis system, also attenuated the [Glu]d elevation following cortical contusion, in the peripheral area of contusion (p < 0.01) but not in the central area. These findings indicate that cerebral contusion involves heterogeneous and complex mechanisms of EAA release into the extracellular space. The release of EAAs in the contusion core was nonsensitive to hypothermia and Co2+ administration, suggesting that such EAA release was related to primary disruption of the cell membrane or vascular wall by the physical force of the head trauma, resulting in leakage of EAAs from the metabolic pool in the cytosole or blood stream. In contrast, in the peripheral area, the effectiveness of hypothermia and Co2+ administration implied a presynaptic mechanism of EAA release, which consisted, at least in part, of Ca2+-dependent exocytotic EAA release from depolarized nerve terminals. The EAAs released in the contusion core may diffuse towards a peripheral direction and act on the postsynaptic receptors, causing neuronal depolarization. Such a diffusion-reaction process appears to induce additional release of EAAs from the depolarized nerve terminals. Hypothermia may block this diffusion-reaction process and eventually reduce the contusion volume. PMID- 9753214 TI - Magnetic resonance spectroscopy of diffuse brain trauma in the pig. AB - The acute metabolic events linked to the evolution of selective axonal pathology in the white matter following diffuse brain injury have not previously been evaluated due to the paucity of relevant experimental models. Here, we utilized a new model of inertial brain injury in the pig that selectively damages axons in the white matter, and applied proton and phosphorous magnetic resonance spectroscopy (MRS) to noninvasively monitor the temporal course of metabolic changes following trauma. Evaluating four pigs with MRS prior to injury, within 1 h and 3 and 7 days postinjury, we found that widespread axonal injury was produced in the absence of changes in pH, PCr/Pi, or the concentrations of ATP, and lactate. However, we did observe an acute 60% loss of intracellular Mg2+ levels, which gradually resolved by 7 days postinjury. In addition, we found that the levels of the neuron marker, N-acetylaspartate (NAA), acutely dropped 20% and remained persistently decreased for at least 7 days postinjury. Moreover, the changes in Mg2+ and NAA were found with MRS in the absence of abnormalities with conventional magnetic resonance imaging (MRI). These results show that (1) profound alterations in intracellular metabolism occur acutely following diffuse axonal pathology in the white matter, but in the absence of indicators of ischemia, and (2) axonal pathology may be evaluated with high sensitivity utilizing noninvasive MRS techniques. PMID- 9753215 TI - Widespread axonal injury in gunshot wounds to the head using amyloid precursor protein as a marker. AB - In order to determine whether axonal injury (AI) is a factor in cases of penetrating head injury, the brains of 14 patients who died shortly after sustaining a fatal gunshot wound (GSW) to the head were examined, and the presence of AI determined using immunohistochemical staining for amyloid precursor protein (APP). The distribution of AI was mapped throughout the cerebral hemispheres and brain stem. AI was present in all cases in a diffuse distribution distant to the missle track with severe involvement of the brain stem in all cases. There was no axonal APP immunoreactivity in the direct region of the missle track at the point of primary axotomy. The APP-positive AI in these cases is likely to be a mixture of primary and secondary AI as APP immunostaining is unable to distinguish primary AI due to mechanical deformation from AI secondary to hypoxic-ischemic damage. PMID- 9753216 TI - Completion rates and feasibility of outcome measures: experience in a multicenter clinical trial of systemic hypothermia for severe head injury. AB - The National Acute Brain Injury Study: Hypothermia (NABIS:H) is an ongoing multicenter trial of systemic hypothermia for the treatment of severe head injury. Follow-up rates for the study's 3-and 6-month outcome assessments have been maintained at high levels by establishing close contact with family members, by reimbursing cost of travel, and by sending examiners to the subject's location whenever necessary. Two years into the study, global disability data (e.g., Glasgow Outcome Scale) have been obtained on 86% of patients due for 3-month assessment (n = 131) and for all subjects due at 6 months (n = 100). Over half of the patients have completed neuropsychological testing with high reliability ratings. These preliminary findings suggest that the procedures used to document data quality and increase follow-up and completion rates are being successful. PMID- 9753217 TI - Infant rat model of the shaken baby syndrome: preliminary characterization and evidence for the role of free radicals in cortical hemorrhaging and progressive neuronal degeneration. AB - Infants subjected to repeated episodes of violent shaking develop brain damage characterized by intracranial hemorrhage and progressive cortical atrophy. We have developed an animal model that mimics this pathological state and investigated its etiology and treatment. Anesthetized male rats, 6 days of age, were subjected to one episode of shaking per day for 3 consecutive days. Separate groups of rats were sacrificed 1 h postinjury on the third day of shaking for HPLC quantification of cortical .OH and vitamin E levels, and histological assessment of cortical hemorrhaging. Additional groups were sacrificed 7 or 14 days postinjury to demonstrate progressive neuronal degeneration via cortical wet weight comparisons. In comparison to noninjured shams, the results indicated that cortical vitamin E and .OH levels rose 53.7% (p < 0.005) and 457.1% (p < 0.001), respectively, in shaken infant rats. Brain histologies revealed a moderate-to severe degree of cortical hemorrhaging in these animals 1 h postinjury. By 7 and 14 days postinjury, there was a 13.3% and 28.7% (p < 0.0001 vs. sham) loss of cortical tissue in shaken infants, respectively, indicating progressive neuronal degeneration. Treatment with 10 mg/kg (ip) of the 21-aminosteroid antioxidant, tirilazad mesylate, 10 min before and 2 h after each episode of shaking, resulted in a 53.1% attenuation of cortical .OH levels and a 34.9% decrease in brain hemorrhaging (p < 0.05 vs. vehicle). Tirilazad treatment did not, however, significantly effect cortical vitamin E concentrations at 1 h postinjury or the extent of progressive neuronal degeneration at either 7 or 14 days postinjury. The present animal model mimics the brain pathology seen in abused children. Our observation that tirilazad mesylate, an antioxidant-lipid peroxidation inhibitor, significantly reduces cortical .OH levels and brain hemorrhaging in shaken infant rats supports a role for oxygen radicals in the pathophysiology of this type of CNS injury. The failure of tirilazad to block progressive cortical degeneration suggests that mechanisms other than free radicals may be of prime importance in the mediation of this aspect of the pathology. PMID- 9753218 TI - Tirilazad widens the therapeutic window for riluzole-induced attenuation of progressive cortical degeneration in an infant rat model of the shaken baby syndrome. AB - Our infant rat model of traumatic subarchnoid hemorrhage combines violent shaking and hypoxia to produce subdural hemorrhaging and progressive cortical degeneration similar to that seen in victims of the shaken baby syndrome. Anesthetized, 6-day-old male rats were subjected to one episode of shaking under hypoxic conditions. Brain histologies revealed moderate-to-severe cortical hemorrhaging at 48 h postinjury and progressive cortical degeneration, as indicated by a 15.3% and 20.2% reduction in cortical wet weight, at 7 and 14 days postinjury, respectively. The purpose of the present study was to assess the effects of two antioxidant lipid peroxidation inhibitors (tirilazad mesylate and PNU-101033E), and the glutamate release inhibitor (riluzole), upon the brain pathology seen in this model. A significant, 54.3-75.3%, reduction in cortical hemorrhaging was observed in rats that were treated with a total of three doses of tirilazad (10 mg/kg, i.p.): 10 min before or 5-30 min after injury, and again at 2 and 24 h postinjury (p < 0.01 vs. vehicle). However, treatment with tirilazad or the more potent, brain-penetrating pyrrolopyrimidine, PNU-101033E (10 min before plus 2, 24, 48, and 72 h after), did not attenuate the progressive cortical degeneration typically seen at 14 days postinjury. These results suggest that free radicals play an important role in the pathophysiology of secondary brain hemorrhaging due to shaking + hypoxia, but may not be critical in the mediation of the subsequent neurodegeneration. Rather, glutamate neurotoxicity may be a key factor here. This is suggested by our observation that the glutamate release inhibitor, riluzole, significantly reduced cortical degeneration when it was administered up to 1 h postinjury in the present model. Specifically, the cortical wet weights of rats treated with 8 mg/kg riluzole (i.p.) 10 min before or 1 h after shaking + hypoxia (and again at 24 h postinjury) were 95.3% and 97.4% of noninjured controls, respectively, at 14 days postinjury (p < 0.02 vs. vehicle). Riluzole treatment beyond 1 h (e.g., 2 or 4 h postinjury) did not reduce the neurodegeneration. Lastly, we attempted to demonstrate that the therapeutic window for riluzole-induced attenuation of cortical degeneration could be extended beyond 1 h through the use of combination therapy. In this experiment, rat pups were treated with 10 mg/kg tirilazad (i.p.) at 30 min postinjury followed by 8 mg/kg riluzole (i.p.) at 4 and 24 h postinjury. At 14 days postinjury, the cortical wet weights of these rats were 94.5% of noninjured controls, thus demonstrating significant neuroprotection (p < 0.05 vs. vehicle) and a widening of the therapeutic window from 1 to 4 h in length. These results suggest that early attenuation of free radical-induced lipid peroxidation may slow down the biochemical cascade of events related to glutamate-induced excitotoxicity and, in doing so, prolong the time during which a glutamate release inhibitor, such as riluzole, is effective. PMID- 9753219 TI - Brain injury impairs prostaglandin cerebrovasodilation. AB - Previous studies have observed that ATP- and calcium-sensitive K+ (KATP and Kca) channel function is impaired after fluid percussion brain injury (FPI). The present study was designed to characterize the effect of FPI on prostaglandin (PG)E2 and 12 pial artery dilation and the role of activation of these K+ channels in that dilation in newborn pigs equipped with a closed cranial window. FPI of moderate severity (1.9-2.1 atm) was produced by using a pendulum to strike a piston on a saline-filled cylinder that was fluid coupled to the brain via a hollow screw inserted through the cranium. PGE2 vasodilation was blunted by FPI (9+/-1%, 13+/-1%, and 19+/-1% vs. 2+/-1%, 5+/-1%, and 9+/-1%, for 1, 10, and 100 ng/ml PGE2 before and after FPI, respectively). PGE2 dilation was associated with increased CSF cGMP and cAMP concentration and such changes in cyclic nucleotides were blunted by FPI (448+/-10 and 793+/-38 vs. 316+/-11 and 403+/-27 fmol/ml for control and PGE2 induced change in cGMP before and after FPI, respectively). PGI2 induced dilation and associated changes in CSF cyclic nucleotide concentration were similarly blunted by FPI. PGE2 dilation was attenuated by either glibenclamide or iberiotoxin, KATP and K,ca channel antagonists, and coadministration of both K+ channel antagonists further decremented the dilator response (9+/-1%, 14+/-1%, and 21+/-1%; vs. 4+/-1%, 7+/-1%, and 12+/-1%; vs. 2+/ 1%, 4+/-1%, and 7+/-1%, for 1, 10, and 100 ng/ml PGE2 during control, after glibenclamide, and after combined glibenclamide and iberiotoxin, respectively). Glibenclamide and iberiotoxin had similar effects on PGI2 dilation. These data show that prostaglandin dilation is attenuated after FPI. These data also show that prostaglandin dilation is dependent on activation of both KATP and Kca channels. Further, these data suggest that attenuated prostaglandin dilation following FPI results from diminished prostaglandin-associated elevation in cyclic nucleotide concentration and impaired KATP and Kca channel function. PMID- 9753220 TI - Rebamipide: overview of its mechanisms of action and efficacy in mucosal protection and ulcer healing. AB - Rebamipide, a gastroprotective drug, is a compound selected from over 500 amino acid analogs of 2(1H)-quinolinone tested for gastroprotective action and for efficacy to heal experimental gastric ulcers. This drug stimulates prostaglandin generation in gastric mucosa and improves not only the speed but also the quality of ulcer healing. In addition, it protects the gastric mucosa against acute injury caused by various noxious and ulcerogenic factors. Based on these experimental results, rebamipide had been subsequently tested in several clinical trials and approved in Japan for therapeutic use in patients with gastric ulcers and patients with acute gastritis. The main purpose of developing this type of drug was to improve the quality of ulcer healing, especially in that antisecretory drugs lack this advantage. In a preliminary clinical study, rebamipide improved the quality of gastric ulcer healing and reduced future ulcer recurrence. A number of basic research studies have been performed to clarify the mechanisms of rebamipide's action. These studies demonstrated unique properties of rebamipide and convincingly showed that it increases gastric mucus glycoprotein components, stimulates migration and proliferation of wounded epithelial cell monolayers, increases expression of epidermal growth factor and its receptor in normal and ulcerated gastric mucosa, and scavenges active oxygen radicals. The drug also attenuates the activity of neutrophils and the production of inflammatory cytokines stimulated by NSAIDs and/or H. pylori. Therefore, rebamipide can contribute to the management of patients who are taking NSAIDs or are infected with H. pylori. The inhibition of immunoinflammatory responses by rebamipide in H. pylori-infected patients may prevent development of gastritis, peptic ulcer disease, its recurrence, and possibly gastric cancer. Moreover, rebamipide may enhance eradication of H. pylori-infection using standard eradication therapy. Further studies are needed to clarify these possible advantages of rebamipide. PMID- 9753221 TI - Signal transduction cascades triggered by EGF receptor activation: relevance to gastric injury repair and ulcer healing. AB - Growth factors and their receptors are known to play important roles in normal cell proliferation, tissue repair, and ulcer healing. Epidermal growth factor (EGF) inhibits acid secretion, protects gastric mucosa against injury, mediates mucosal adaptation, and accelerates gastroduodenal ulcer healing. EGF exerts its actions by binding to its receptor (EGF-R), which is a transmembrane protein tyrosine kinase. Binding of EGF to its receptor triggers receptor dimerization and autophosphorylation, recruitment of kinase substrates (signaling enzyme adapter proteins with an SH2 domain, Grb2 adapter protein, and Grb2-SOS complex). These events lead to Ras (GTP-binding protein) phosphorylation and activation of the Ras/Raf/MAP kinase pathway, in turn leading to phosphorylation of regulatory proteins and transcription factors and culminating in cell proliferation. Other pathways potentially activated by EGF include the phosphatidylinositol pathway (leading to activation of protein kinase C and an increase in cytosolic calcium) and the JAK/STAT signaling pathway. While EGF-induced signaling events have been extensively studied in various cell systems, predominantly neoplastic and/or transformed cells, the relevance of those findings to gastric mucosal injury repair or ulcer healing is as yet not fully elucidated. This paper is intended to provide an overview of signaling pathways triggered by EGF-R activation and on this background to summarize current knowledge pertaining to involvement of EGF-R signaling pathways in gastric mucosal repair and ulcer healing. PMID- 9753222 TI - Role of prostaglandins in gastroprotection. AB - Numerous agents increase gastric mucosal resistance against intraluminal ulcerogens. Although the precise mechanisms of gastroprotection are uncertain, various endogenous mediators involved in gastroprotective effects have been characterized. As prostaglandins exert potent protective effects and inhibition of prostaglandin formation abolishes "adaptive gastroprotection," they have been proposed as key mediators in mucosal defense. This paper reviews the role of endogenous prostaglandins showing striking differences between different forms of gastroprotection. Thus, whereas the protective effect of the antiulcer drug rebamipide involves prostaglandins as essential mediators, the protection conferred by the antacid hydrotalcit is prostaglandin-independent. Furthermore, gastroprotection can occur even when mucosal prostaglandin generation is suppressed. This phenomenon has been observed with some nonsteroidal antiinflammatory drugs, agents that modulate sulfhydryls and certain metals. Recent data suggest that both cyclooxygenase-1- and cyclooxygenase-2-derived prostaglandins can increase mucosal resistance. The precise role of constitutive and inducible forms of cyclooxygenase in gastroprotection, however, remains to be established. PMID- 9753223 TI - Role of oxygen radical and lipid peroxidation in indomethacin-induced gastric mucosal injury. AB - Nonsteroidal antiinflammatory drugs such as aspirin and indomethacin are known to induce gastric mucosal damage including bleeding, ulceration, and perforation in humans and animals. Although it has been proposed that a deficiency of endogenous prostaglandins due to inhibition of cyclooxygenase by the drug is involved in these effects, the exact pathogenic mechanism remains to be elucidated. It has recently been proposed that neutrophil- and oxygen radical-dependent microvascular injuries may be important prime events that lead to mucosal injury induced by nonsteroidal antiinflammatory drugs. Lipid peroxidation mediated by oxygen radicals, especially hydroxyl radicals, plays a crucial role in the development of the gastric mucosal injury induced by indomethacin. Both allopurinol, an inhibitor of xanthine oxidase, and neutrophil depletion by intraperitoneal injection of antineutrophil antibody significantly attenuates indomethacin-induced gastric injury. In this paper, we have reviewed the recent data that assess the role of oxygen radical and lipid peroxidation in the pathogenesis of indomethacin-induced gastric mucosal injury in rats and humans. PMID- 9753224 TI - In vitro studies indicating antioxidative properties of rebamipide. AB - Rebamipide is the first anti-gastric ulcer and antigastritis drug that not only increases endogenous prostaglandin in gastric mucosa but also scavenges oxygen derived free radicals and inhibits their production. In the present paper, we have reviewed the antioxidative and antiinflammatory properties of rebamipide mainly demonstrated by in vitro studies. The study, using the electron paramagnetic resonance (EPR) spin trapping technique, showed that superoxide production was inhibited by rebamipide when isolated human neutrophils were stimulated with opsonized zymosan or Helicobacter pylori water extract in a dose dependent manner. Chemiluminescence generated from neutrophils activated by H. pylori or formyl-methionyl-leucyl-phenylalanine was also decreased by the treatment with rebamipide. Rebamipide, at concentrations of 10(-5) and 10(-6) M, reduced the adherence of neutrophils to endothelial cells as well as the CD18 expression on neutrophils induced by H. pylori water extract. The EPR study also demonstrated the direct hydroxyl radical scavenging activity of rebamipide, and a kinetic study showed that the second-order rate constant for the reaction between rebamipide and hydroxyl radical was 2.24 x 10(10) M(-1)/s(-1). The inhibitory effect of rebamipide on lipid peroxidation induced by a free radical initiator was also demonstrated by the in vitro system using rat gastric mucosal homogenates. These data indicate that rebamipide offers a potential for protection against reactive oxygen- and activated neutrophil-associated gastric mucosal injury by scavenging hydroxyl radical and inhibiting neutrophil activation or lipid peroxidation. PMID- 9753225 TI - Vascular approach to gastroduodenal ulceration: new studies with endothelins and VEGF. AB - Endothelins (ET) and VEGF/VPF (vascular endothelial growth factor/vascular permeability factor) are products mainly of endothelial cells, which are also regulated via autocrine and paracrine pathways by these peptides. As a follow-up to our focus on vascular factors in ulcer pathogenesis and healing, we review here our recent studies with ET-1 and VEGF/VPF in animal models and human subjects. Our new results demonstrated a rapid and time-dependent release of ET-1 into the systemic circulation after intragastric administration of ethanol or HCI in rats, and ethanol in humans. The ET-1 release preceded the development of hemorrhagic erosions in both species and might be used as a diagnostic tool to noninvasively quantify acute gastric mucosal lesions. The development of solitary duodenal ulcers in the rat was preceded only by an organ- (involving only the duodenum and not the stomach) and molecule-specific (induced only by cysteamine and not by the nonulcerogenic analog ethanolamine) rapid local release of ET-1. The severity of cysteamine-induced duodenal ulcers was dose-dependently decreased by pretreatment with ET-1 antibodies or antagonist bosentan. A single intragastric dose of VEGF/VPF resulted in gastroprotection against ethanol, while daily intragastric treatment with the peptide for three weeks stimulated angiogenesis in the base of cysteamine-induced duodenal ulcers and accelerated ulcer healing. Thus, modulation of vascular factors seems to be sufficient for both acute gastroprotection and chronic duodenal ulcer healing. PMID- 9753226 TI - Role of cytokines in pathogenesis of Helicobacter pylori-induced mucosal damage. AB - In Helicobacter pylori infection both bacterial and host factors contribute to gastroduodenal mucosal damage. Indirect damage will result from the persistent innate and specific inflammatory response induced by bacterial products and the alterations in gastric physiology associated with infection. Cytokines play a critical role in the initiation and modulation of gastrointestinal inflammation. The gastric epithelium, which secretes chemokines in response to H. pylori, has a role in initiating acute inflammation. Bacterial induction of epithelial chemokines such as IL-8 involves protein tyrosine phosphorylation and NF-kappaB activation. Multiple genes in the cag pathogenicity island are essential for induction of epithelial chemokines. In vivo infection with cag-positive strains is associated with increased mucosal chemokines and inflammatory responses. Th1 cell-mediated responses characterized by interferon-gamma-secreting effector cells are also associated with increased mucosal inflammation. Variations in the magnitude and characteristics of the host cytokine responses induced by H. pylori are considered important factors determining the degree of chronic inflammation. PMID- 9753227 TI - Modification of Helicobacter pylori adhesion to human gastric epithelial cells by antiadhesion agents. AB - Helicobacter pylori is a major etiological agent in gastroduodenal disorders. H. pylori adhesion to human gastric mucosa is the initial step of H. pylori colonization. Inhibition of H. pylori adhesion is thus a therapeutic target in preventing H. pylori infection. We have previously established a method using the enzyme-linked immunosorbent assay to analyze quantitatively H. pylori adhesion to gastric epithelial cells. This method is suitable for screening antiadhesion agents. Some mucoprotective agents are proved to have antiadhesion effects in vitro, and they may modify H. pylori adhesion. This evidence gives us a useful clue to analyze the molecular mechanism of H. pylori adhesion to mucosa. Furthermore, in clinical trials, these mucoprotective agents enhanced the eradication rate when administered with antibiotics. In conclusion, the antiadhesion agents may have potential as therapeutic regimens against H. pylori infection. PMID- 9753228 TI - Ulcer recurrence: cytokines and inflammatory response-dependent process. AB - H. pylori and nonsteroidal antiinflammatory drugs (NSAIDs) are important factors in the recurrence of peptic ulcer diseases. However, H. pylori-negative recurring ulcers can also be found in nonusers of NSAIDs. The aim of this paper is to review recent data pertaining to mechanisms of ulcer recurrence. Prostaglandin E2 generation is impaired in the tissues of the ulcer scar site and prostaglandin depletion induced by administration of indomethacin during the healing of experimental gastric ulcer predisposes to future ulcer recurrence. Therefore, the prostaglandin deficiency may impair the quality of ulcer healing and thus increase the likelihood of future ulcer recurrence. Persistent infiltration of polymorphonuclear cells is the most prominent finding in the gastric ulcer scar in rats treated with indomethacin. Concomitant administration of prostaglandin E1 analog with indomethacin attenuates inflammatory infiltration and reduces future ulcer recurrence. Therefore, the inflammatory responses at the ulcer scar site may be a key to the quality of ulcer healing. Recent clinical findings suggest a close relationship between the quality of ulcer healing, infiltration of neutrophils and mononuclear cells, and future ulcer recurrence. Gastroprotective drugs such as prostaglandin analogs and prostaglandin inducers improve the quality of ulcer healing and reduce future recurrence. Production of inflammatory cytokines is stimulated by ulcerogenic factors such as NSAIDs, stress, and H. pylori infection. Inflammatory cytokines such as interleukin-1beta and tumor necrosis factor-alpha cause recurrence of healed ulcer. Synthetic prostaglandin E2 inhibits recurrence as well as the production of the cytokines. PMID- 9753229 TI - Nonulcer dyspepsia and Helicobacter pylori, with comment on posteradication symptoms. AB - In two London hospitals during five months in 1997, among patients referred for esophago-gastroduodenoscopy, 250 complained of dyspepsia for more than two days per week. Of these, 190 gave informed consent to enter a study of H. pylori infection in nonulcer dyspepsia, but only 42 (22%) were found to have H. pylori infection without a peptic ulcer. At the time of this interim report, of these patients, 26 had been treated with omeprazole, amoxicillin, and clarithromycin, four weeks had elapsed since treatment, and H. pylori had been eradicated. Of these 26 patients, 15 (58%) had lost nearly all their symptoms. This is the first report of loss of symptoms in patients with nonulcer dyspepsia after treatment with omeprazole, amoxicillin and clarithromycin with early follow-up after four weeks. However, this was not a placebo-controlled study and the number of patients was small, so it is not possible to conclude whether H. pylori could be one cause of nonulcer dyspepsia. The increasing incidence of posteradication esophagitis is discussed as is the possible need for more sophisticated management of nonulcer dyspepsia. PMID- 9753230 TI - Helicobacter pylori infection, oxidative DNA damage, gastric carcinogenesis, and reversibility by rebamipide. AB - Several epidemiological studies have demonstrated a close association between Helicobacter pylori infection and carcinoma of the mid- or distal stomach. If this can be shown to be a causal association, eradication of the organism may prevent later development of cancer. Several mechanisms have been proposed by which H. pylori infection might lead to predisposition for gastric cancer. Although many potential pathogenic mechanisms, such as increased proliferative gastric epithelial response to H. pylori, lowered gastric ascorbic acid levels, and high occurrences of atrophic gastritis, have been proposed, there is little evidence as to which might be of direct importance to such H. pylori-related disease in vivo. H. pylori-associated inflammation may interact with other causal factors related to gastric carcinogenesis and can result in the intestinal type of gastric cancer and then DNA damage due to oxygen radicals induced by persistent inflammatory cell infiltrations in the gastric mucosa may lead to alterations of the gene and result in the development of diffuse-type carcinoma. In order to know the influence of H. pylori on changes of inflammation-related DNA damage, we measured the sequential changes of 8-hydroxydeoxyguanosine (8 OHdG) contents of DNA and the changes of two biomarkers inducible nitric oxide synthase (iNOS) and apoptosis from human gastric mucosa according to the status of H. pylori. The increased levels of oxidative DNA damage, increased occurrences of apoptosis, and increased expressions of iNOS seem to provide the mechanistic links between H. pylori infection and gastric carcinogenesis and rebamipide can abrogate the levels of these hazard factors. PMID- 9753231 TI - Protective effect of rebamipide against ammonia-induced gastric mucosal lesions. AB - We investigated the protective effect of rebamipide against ammonia-induced gastric mucosal lesions. Participation of prostaglandin E2 and nitric oxide in the action of rebamipide was also examined. Rebamipide was administered intraperitoneally (10-100 mg/kg) to male Wistar/ST rats (150-325 g) fasted for 24 hr. Thirty minutes later, 1% NH4OH (1 ml) solution was given intragastrically. One hour later, the length of the mucosal lesions was measured (lesion index), and prostaglandin E2 (PGE2) was determined by radioimmunoassay. A 1% NH4OH solution caused gastric mucosal lesions with hemorrhagic necrosis and submucosal edema. PGE2 synthesis was not affected by NH4OH but was significantly increased by rebamipide. Rebamipide decreased the severity of NH4OH-induced gastric mucosal lesions in a dose-dependent manner. Pretreatment with indomethacin (5 mg/kg, subcutaneously) did not affect the protective effect of rebamipide; however, pretreatment with N(omega)-nitro-L-arginine (L-NNA, 1-10 mg/kg, intravenously), an inhibitor of nitric oxide synthase, attenuated the protective effect of rebamipide in a dose-dependent manner. Simultaneous administration of L-arginine (100 mg/kg) and L-NNA completely restored the protective effect of rebamipide, whereas D-arginine was inactive. These results suggest that nitric oxide contributes significantly to the protective effect of rebamipide against ammonia induced gastric mucosal lesions. PMID- 9753233 TI - Rebamipide treatment activates epidermal growth factor and its receptor expression in normal and ulcerated gastric mucosa in rats: one mechanism for its ulcer healing action? AB - Rebamipide (Mucosta) is a novel mucosal protective and ulcer-healing drug. Clinical and experimental data indicate that it accelerates ulcer healing and improves the quality of the scar. Since epidermal growth factor (EGF) and its receptor (EGF-R) play important roles in mucosal protection and ulcer healing, we studied whether rebamipide treatment affects expression of EGF and EGF-R in normal and ulcerated gastric mucosa in rats. Forty-eight male rats without or with gastric ulcers (induced by acetic acid) received intragastrically either placebo or rebamipide, 40 mg twice daily, for 14 days. Ulcer size was measured under a dissecting microscope; mucosal sections were stained with H&E or immunostained with specific antibodies against EGF and EGF-R. The distribution and intensity of fluorescence signal was quantified using a video image system. Rebamipide significantly accelerated ulcer healing, produced a significant increase in EGF and EGF-R expression in normal gastric mucosa (both P < 0.001), and increased expression of EGF and EGF-R in regenerating glands of the ulcer scar. Since EGF and its receptor are crucial for epithelial cell proliferation, re-epithelialization, and gland reconstruction, the above actions of rebamipide may provide a new mechanism for its ulcer healing action. PMID- 9753232 TI - Rebamipide protects against indomethacin-induced gastric mucosal injury in healthy volunteers in a double-blind, placebo-controlled study. AB - The aim of the present study was to evaluate rebamipide in the prevention of indomethacin-induced gastric mucosal injury in healthy volunteers. Twenty healthy males (mean age 21.8 years, range 20-26) participated. This is a randomized, double-blind, placebo-controlled study. The 20 subjects were randomized to either indomethacin 25 mg three times a day and placebo three times a day or indomethacin and rebamipide 100 mg three times a day for seven days. Endoscopy was performed at baseline and again after the treatment. In the placebo group, eight of 10 subjects (80%) developed symptoms compared to three of seven (43%) in the rebamipide group. The incidence of gastric lesions was 70% in the placebo group, which was significantly higher than that in the rebamipide group (14%). The lipid peroxide levels in the mucosa of the gastric body significantly increased in the placebo group. This increase was not inhibited by rebamipide. Myeloperoxidase activity in the gastric mucosa tended to increase in the placebo group, but tended to decrease in the rebamipide group. These results indicate that rebamipide may be an effective prophylaxis against indomethacin-induced gastropathy in humans. PMID- 9753234 TI - Rebamipide prevents recurrence of gastric ulcers without affecting Helicobacter pylori status. AB - Rebamipide, a gastroprotective drug developed in Japan, accelerates ulcer healing and reduces recurrence of experimental gastric ulcers. We examined the effects of rebamipide, given during healing of human gastric ulcers infected with Helicobacter pylori, on the quality of ulcer healing and ulcer recurrence. Sixty H. pylori-positive patients with gastric ulcers were randomly allocated to three treatment groups: group O (N = 20) received 20 mg of omeprazole every day for eight weeks, group OR (N = 20) received the same dose of omeprazole and 300 mg of rebamipide three times a day for eight weeks, and group OA (N = 20) received the same dose of omeprazole for eight weeks and 1500 mg of amoxicillin three times a day for the first two weeks. After this treatment was completed no other medication was given. Endoscopic examinations were performed at the end of therapy (for healing rate), one month later (for rate of H. pylori eradication) and every three months for follow-up (for ulcer recurrence rate). At the end of therapy, biopsy specimens were taken from the gastric ulcer scar and examined under the microscope for neutrophil and mononuclear cell infiltration. The ulcer healing rate of the three groups was almost the same; H. pylori in group OA was 65% and that of the other two groups was 0%. The number of patients with a flat ulcer scar pattern (good quality of ulcer healing) was increased and the neutrophil infiltration was significantly improved in groups OR and OA compared to group O. The ulcer recurrence rate was significantly lower in group OA and group OR than in group O. In conclusion, rebamipide is almost equipotent to amoxicillin plus omeprazole for the reduction of ulcer recurrence. The decreased recurrence rate by rebamipide may be due to improvement of the quality of ulcer healing, reflected as in the suppression of inflammatory cell infiltration in the scar, which results from either cure of H. pylori infection and/or treatment with a gastroprotective drug such as rebamipide. PMID- 9753235 TI - Rebamipide prevented delay of wound repair induced by hydrogen peroxide and suppressed apoptosis of gastric epithelial cells in vitro. AB - The effects of rebamipide on the restoration process of gastric epithelial wounds were assessed using an in vitro wound-healing model and hydrogen peroxide treatment. Rebamipide (10 or 100 microM) was added to a complete monolayer cell sheet after artificial wounding. The restoration process was analyzed sequentially by a time-lapse video system, and cell migration, proliferation, and apoptosis were assessed. Hydrogen peroxide (1 or 3 mM) inhibited restoration after wounding by suppressing cell migration and proliferation. It also induced epithelial cell apoptosis around the wound. The addition of rebamipide abolished the H2O2-induced retardation and prevented apoptosis. Rebamipide might act as a radical scavenger and have favorable effects on peptic ulcer diseases. PMID- 9753236 TI - Rebamipide protects against oxygen radical-mediated gastric mucosal injury in rats. AB - Rebamipide, a novel antiulcer agent, has been shown to protect against gastric injury by free radicals. The effect of rebamipide was examined using two rat models of mucosal injury: the stomach was exposed to luminal perfusion of 10 mM H2O2 for 10 min or to local ischemia for 30 min. The effect of deferoxamine, a chelator of Fe3+, was also evaluated to determine whether Fe3+-mediated production of hydroxyl radicals contributed to the damage induced by H2O2. The pylorus was ligated and a double-lumen cannula was inserted into the forestomach for luminal perfusion. [51Cr]EDTA was administered intravenously and mucosal integrity was monitored by measuring blood-to-lumen [51Cr]EDTA clearance. Rebamipide reduced the increase in EDTA clearance induced by ischemia or H2O2. Furthermore, deferoxamine attenuated the H2O2-induced increase. These results suggest that rebamipide has a protective effect against oxygen radical-mediated gastric damage and that Fe3+ is involved in the H2O2-induced injury. PMID- 9753237 TI - IFN-gamma-induced iNOS mRNA expression is inhibited by rebamipide in murine macrophage RAW264.7 cells. AB - To investigate the effects of rebamipide, an antigastritis and anti-gastric ulcer drug, on inducible nitric oxide synthase (iNOS), murine macrophage RAW264.7 cells were treated with interferon-gamma (IFN-gamma) in the presence of rebamipide. NO production was stimulated by IFN-gamma, and the level was attenuated by rebamipide in a dose-dependent manner. Therefore, we investigated the possibility that either rebamipide directly inhibited iNOS enzyme activity or that it reduced iNOS mRNA expression. In a cell-free system, rebamipide did not affect iNOS enzyme activity; however, rebamipide inhibited iNOS mRNA and protein expression induced by IFN-gamma. Thus, we concluded that rebamipide inhibited IFN-gamma induced NO production as a result of its inhibitory action on iNOS mRNA expression. PMID- 9753238 TI - Protection by rebamipide against acetic acid-induced colitis in rats: relationship with its antioxidative activity. AB - The aim of this study was to examine the efficacy of rebamipide on acetic acid induced colitis in rats. Colitis was induced in male Wistar/ST strain rats by intracolonic injection of 2 ml of 5% acetic acid. After 24 hr, lesion score, myeloperoxidase (MPO) activity, thiobarbituric acid-reactive substances (TBARS), and antioxidants were measured. Rebamipide was administered orally to the rats twice, once immediately and once 8 hr after the induction of the colitis at doses of 7.5, 15, and 30 mg/kg. To evaluate the pathogenesis of neutrophils in this colitis model, neutrophil-depleted rats were also investigated. Increases of TBARS and MPO activity and decreases of total glutathione, superoxide dismutase (SOD) activity, and alpha-tocopherol were observed in this model. Rebamipide at 30 mg/kg decreased the lesion score significantly, which was associated with the reduction of colonic TBARS. The decreases of total glutathione, SOD activity, and alpha-tocopherol were also inhibited significantly by rebamipide. The preventive effect of rebamipide in this colitis model may be attributed to its inhibition of reactive oxygen species production. PMID- 9753240 TI - Rebamipide prevents indomethacin-induced gastric mucosal lesion formation by inhibiting activation of neutrophils in rats. AB - Since granulocyte elastase has been shown to be involved in the pathogenesis of gastric mucosal lesion formation induced by nonsteroidal antiinflammatory drugs, inhibition of granulocyte elastase release from neutrophils may be useful in the prevention of these lesions. The objective of this study was to determine whether rebamipide inhibits neutrophil activation in vivo and in vitro. Rebamipide and ONO-5046, a specific granulocyte elastase inhibitor, markedly inhibited indomethacin-induced mucosal injury in rats. Gastric myeloperoxidase activity was significantly increased 3 h after indomethacin administration. This increase was significantly inhibited by rebamipide and ONO-5046. Although cimetidine markedly prevented the indomethacin-induced mucosal lesion formation, it did not reduce the gastric myeloperoxidase activity. Rebamipide inhibited granulocyte elastase release from neutrophils in vitro by inhibiting the increase in intracellular Ca2+ level. Cimetidine did not inhibit granulocyte elastase release from neutrophils. Furthermore, the elevation of intracellular Ca2+ level was not inhibited by cimetidine. Therefore, unlike cimetidine, rebamipide may prevent indomethacin-induced mucosal injury by inhibiting neutrophil activation. PMID- 9753239 TI - Increased mRNA levels of transforming growth factor-beta1 and monocyte chemoattractant protein-1 in ulcer relapse caused by interleukin-1beta in rats. AB - This study investigated the mRNA expression of transforming growth factor-beta1 (TGF-beta1) and monocyte chemoattractant protein-1 (MCP-1) in rat gastric tissues in which ulcers had relapsed due to interleukin-1beta (IL-1beta) administration. Rats with healed ulcers were administered IL-1beta (1 microg/kg) and killed after 0, 12, 24, or 48 hr. Both TGF-beta1 and MCP-1 mRNA levels were increased in the scarred gastric tissues at 24 hr (fourfold), when ulcers had not relapsed. Furthermore, the expression of these genes also increased in the ulcerated gastric tissues at 48 hr (fivefold), when 90% of healed ulcers had relapsed. On the other hand, the number of macrophages that had infiltrated the scarred gastric tissues at 24 hr was two times higher than that at 0 hr. At 48 hr, the number of macrophages that had infiltrated gastric tissues in which ulcers had relapsed was similar to that at 24 hr. Thus, TGF-beta1 and MCP-1 may be implicated in the macrophage infiltration, thereby leading to ulcer relapse due to IL-1beta. PMID- 9753241 TI - Changes in colonic inflammation induced by dextran sulfate sodium (DSS) during short- and long-term administration of rebamipide. AB - Earlier studies have shown the antiinflammatory effects of histamine and nitric oxide (NO) in a model of colitis induced by DSS. However, the defense system against free radicals in this model remained unclear. The aim of this study was to evaluate the effects of rebamipide, which inhibits the production of free radicals, in this model using male Sprague-Dawley rats. Colitis induced by 1% DSS is characterized by slow, weak inflammation and is regarded as a chronic inflammation model. In contrast, colitis induced by 4% DSS is characterized by fast, strong inflammation and is regarded as the acute inflammation model. Endoscopic examinations, peripheral white blood cell (WBC) counts, and assays of myeloperoxidase activity (MPO) in homogenates of colon mucosa were performed after one week (4% DSS model) and eight weeks (1% DSS model). Inflammation of colon mucosa was milder in the rats given rebamipide compared with controls in both the 4% and 1% DSS model. Furthermore, peripheral WBC counts correlated with colonic MPO activity. These findings indicate that rebamipide works as an antiinflammatory agent in both acute and chronic inflammation. PMID- 9753242 TI - Rebamipide attenuates gastric microcirculatory disturbances in the early period after thermal injury in rats. AB - In our previous study we showed that rebamipide attenuated gastric erosions and active oxygen species released only from the gastric mucosa and not from circulating leukocytes after thermal injury. This study was designed to examine whether rebamipide affects the potential of active oxygen generation from circulating leukocytes, and attenuates microcirculatory disturbance caused by thermal injury to skin. Rats were anesthetized and a 30% full skin-thickness dorsal burn was inflicted. Microvascular images and leukocytes were observed using in vivo microscopy. Endothelial damage was assessed by monastral blue B deposits. Active oxygen species were measured by the chemiluminescence method. Rebamipide (100 mg/kg) decreased leukocyte rolling and monastral blue B deposits in venules but did not improve arteriolar contractions 15 min after thermal injury. These results suggest that rebamipide preserves gastric microcirculation possibly through inhibition of leukocyte adhesion and endothelial damage caused by thermal injury to skin. PMID- 9753243 TI - Effect of rebamipide on liver damage and increased tumor necrosis factor in a rat model of endotoxin shock. AB - We investigated the effect of rebamipide, a novel antiinflammatory agent, on liver damage in a rat model of circulatory shock induced by bacterial endotoxin (E. coli lipopolysaccharide, LPS). Endotoxemia for 6 hr resulted in a 5.9-fold rise in the serum levels of nitrite (P < 0.05) with a significant rise in the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), and lactic dehydrogenase (LDH), suggestive of liver dysfunction. The increased activities of serum ALT, AST, and LDH, but not serum nitrite were significantly inhibited by rebamipide (100 mg/kg, orally for five days). Myeloperoxidase activity in the liver was significantly elevated in the rats with endotoxemia by 2.4-fold (P < 0.05), which was also significantly inhibited by rebamipide. Upon LPS injection, serum TNF-alpha levels peaked at 1 hr after LPS (from 167.4 +/- 20.0 to 1570.0 +/- 100.0 pg/ml) and thereafter rapidly declined. The increased TNF-alpha level measured at 1 hr was significantly inhibited by pretreatment with rebamipide (100 mg/kg for five days). It is suggested that rebamipide exerts a strong protective effect on the LPS-induced liver damage through inhibition of activation of neutrophils and TNF-alpha production. PMID- 9753244 TI - Effects of rebamipide on production of several cytokines by human peripheral blood mononuclear cells. AB - Recently, the relative contributions of local T helper cell responses of the Th1 type and Th2-type to the pathogenesis of gastritis and peptic ulcers associated with Helicobacter pylori infection have been examined. However, the results were controversial with respect to whether cellular immunity (Th1-type) or humoral immunity (Th2-type) responses predominate in H. pylori infection and with respect to how these responses may contribute to disease pathogenesis. In this study, we investigated the characteristics of the production of various cytokines induced by H. pylori or lipopolysaccharide (LPS), which was derived from H. pylori or Escherichia coli, in human peripheral blood mononuclear cells (PBMC). Live H. pylori induced production of many cytokines, such as IL-1beta, IL-10, IL-8, IFN gamma, and TNF-alpha, whereas we could not detect IL-2 or IL-4. Moreover, we evaluated the effect of rebamipide on the production of several cytokines from PBMC induced by various stimuli. Rebamipide suppressed the production of IL-8, IL 10, TNF-alpha, and IL-1beta induced by H. pylori in a dose-dependent manner. On the other hand, the production of IL-12 induced by H. pylori showed a tendency to increase as a result of treatment of the cells with rebamipide. These results suggested that rebamipide might be effective in regulating cytokine responses in the H. pylori-infected host and maintaining host immunity. Moreover, it might contribute positively to disease progression and bacterial eradication. PMID- 9753245 TI - Nonopsonic activation of neutrophils by Helicobacter pylori is inhibited by rebamipide. AB - Some clinical isolates of nonopsonized H. pylori have the ability to activate neutrophils to an oxidative burst (neutrophil activating capacity, NAC), and such strains were significantly more often isolated from patients with peptic ulcer disease and active chronic gastritis. The purpose of the present work was to investigate the effect of rebamipide (Mucosta) on the release of reactive oxygen metabolites from neutrophils activated by various strains of H. pylori with or without NAC, nonopsonized or opsonized, using as controls fMLP and PMA, known activators of neutrophils, and to study the kinetics of these events by luminol enhanced chemiluminescence and by flow cytometry. The results showed that the oxidative burst induced in neutrophils by fMLP and by nonopsonized or opsonized H. pylori with NAC was inhibited by rebamipide in a dose-dependent manner both in the early and late phases of activation. In contrast, the oxidative burst induced by opsonized H. pylori without NAC was not inhibited by rebamipide, which might indicate that it does not have the ability to block CR1 or CR3 receptors involved in opsonic phagocytosis but still has the ability to block the receptor(s) for NAC. The oxidative burst induced by PMA, which primarily activates protein kinase C, was not inhibited in the early phase but diminished 40-45% in the late phase with the 2 mM concentration of rebamipide, probably due to scavenging of reactive oxygen species. In conclusion, rebamipide has the ability to diminish the oxidative burst of neutrophils activated by nonopsonized or opsonized H. pylori organisms with neutrophil activating capacity, most likely through the blocking of fMLP-related receptors, inhibition of the production of reactive oxygen species, and the scavenging of such metabolites. Rebamipide may therefore be useful to prevent gastroduodenal lesions associated with gastric mucosal inflammation in H. pylori infection. PMID- 9753246 TI - Molecular analysis of suppression of interleukin-8 production by rebamipide in Helicobacter pylori-stimulated gastric cancer cell lines. AB - Interleukin-8 (IL-8) may play an important role in Helicobacter pylori infection associated chronic active gastritis and peptic ulcer disease in human. We have recently reported that a gastric cancer cell line, MKN45, produced a massive amount of IL-8 upon coculture with live H. pylori. Moreover, H. pylori induced the activation of NF-kappaB as well as AP-1, leading to IL-8 gene transcription. In this study, we evaluated the effect of rebamipide, an antigastritis and antiulcer agent, on H. pylori-induced IL-8 production. Rebamipide inhibited the production of IL-8 in several gastric cancer cell lines infected with H. pylori. In addition, rebamipide suppressed H. pylori-induced IL-8 gene expression at the transcriptional level as revealed by northern blotting analysis and luciferase activity in cells that were transfected with a luciferase expression vector linked with a 5'-flanking region of the IL-8 gene (bp -133 to +44). Furthermore, rebamipide significantly suppressed the NF-kappaB activation by H. pylori infection. These results suggest that rebamipide may protect against the mucosal inflammation associated with H. pylori infection through inhibition of a proinflammatory cytokine, IL-8. PMID- 9753247 TI - Effect of rebamipide on H. pylori-associated gastric mucosal injury in Mongolian gerbils. AB - Helicobacter pylori colonized to gastric mucosa plays an important pathogenic role in gastric mucosal lesions. We previously reported that ethanol pretreatment promotes the extension of H. pylori-associated lesions. The present study was designed to examine the effect of rebamipide, a mucosal protective agent, on H. pylori-associated injury. Male Mongolian gerbils were orally inoculated with H. pylori; 30 min prior to inoculation, 40% ethanol was administered orally to these gerbils (Hp group). Controls were given 40% ethanol with culture medium (control group). Some gerbils in the Hp and control groups were fed rebamipide-containing diets, and the remaining gerbils received laboratory chow diets. H. pylori infection was evaluated by quantitative bacterial culture and histological examination. Although H. pylori was persistently detected and a remarkable mucosal leukocyte infiltration was observed in the Hp groups, the bacteria had disappeared naturally in 67% of the gerbils and mucosal damage was mitigated in the Hp + rebamipide group at four weeks after the inoculation. Collectively, rebamipide might play a role in inhibiting the level of H. pylori colonization and gastric lesion formation in Mongolian gerbils. PMID- 9753248 TI - Distinction of the shape of Helicobacter pylori using stereo pairs constructed from digitized light microscopic images. AB - This study was performed to distinguish between the coccoid form or spiral forms of Helicobacter pylori (Hp) and to elucidate the pathologic significance of these shapes of Hp. Specimens obtained from human or Mongolian gerbil stomachs were fixed in Carnoy's solution and embedded in paraffin. Sections 3 or 10 microm thick were stained with polyclonal anti-Hp antibody by the immunoperoxidase method. Stereo pairs were prepared from these thick sections by computer-assisted reconstruction. The two shapes of Hp were easily distinguishable by this method. In the human stomach, the proportions of Hp in the surface mucous gel layer (SMGL) and the gastric pits were 31.5% and 68.5%, and the percentage of the spiral form varied from 31.9% to 66.3%. In Mongolian gerbils, a higher proportion of Hp colonized the SMGL, and the spiral form existed more frequently both in the SMGL and on the surfaces of the surface mucous cells. PMID- 9753249 TI - Quantitative and qualitative usefulness of rebamipide in eradication regimen of Helicobacter pylori. AB - The aim of the present study was to determine the efficacy of a new combination regimen including antioxidant, proton pump inhibitor, and antibiotics against Helicobacter pylori and to document the changes of oxidative stress and cytokines involved in H. pylori-associated gastritis. From each of 57 patients with endoscopically diagnosed gastric and/or duodenal ulcers associated with H. pylori infection, five gastric antral biopsy specimens were taken for the diagnosis of H. pylori and for experimental measures. The patients were then treated either with lansoprozole 30 mg + amoxicillin 1.5 g (LA group; 21 patients) or lansoprazole 30 mg + amoxicillin 1.5 g + rebamipide 300 mg (LAM group; 36 patients) for two weeks. Four weeks after the initiation of treatment, the patients were endoscoped again and biopsy specimens were obtained. Mucosal malondialdehyde (MDA) levels; myeloperoxidase (MPO) activities; superoxide dismutase; catalase; glutathione peroxidase; cytokines IL-1, IL-6, TNF-alpha; and chemokines IL-8, GRO-alpha, RANTES (regulated on activation normal T expressed and secreted) were measured. Using paraffin-embedded tissue sections, in situ terminal deoxyribonucleotide transferase (TdT) -mediated dUTP nick end labeling (TUNEL) for apoptosis and immunohistochemical staining for inducible nitric oxide synthase (iNOS) were performed. Two weeks of treatment with the LA regimen resulted in 57.4% eradication rates of H. pylori, whereas two weeks of treatment with the LAM regimen resulted in 75.0% eradication rates. Eradication rates between these two groups were statistically significantly different (P < 0.05). Mucosal MDA levels and MPO activities were significantly lower in the LAM group than the LA group. Mucosal levels of cytokines IL-1, IL-6, and TNF-alpha and of chemokines IL-8, GRO-alpha, and RANTES were all significantly decreased after the treatment of H. pylori, especially in the LAM-treated group. The apoptotic index and iNOS score were significantly reduced after the eradication of H. pylori. The addition of the antioxidative drug rebamipide to the eradication regimen against H. pylori has quantitative and qualitative advantages such as either augmenting the eradication rates of H. pylori or decreasing oxidative stress and cytokines levels generated by H. pylori infection. PMID- 9753250 TI - Effects of rebamipide in combination with lansoprazole and amoxicillin on Helicobacter pylori-infected gastric ulcer patients. AB - The aim of this study was to compare the additive effect of rebamipide with that of teprenone in combination with dual therapy on H. pylori eradication. A total of 102 H. pylori-positive gastric ulcer patients were assigned at random to two groups; in addition to dual therapy (amoxicillin 500 mg thrice daily and lansoprazole 30 mg every morning for two weeks), one group received rebamipide 100 mg thrice daily for eight weeks, while the other group received teprenone 50 mg thrice daily for eight weeks. H. pylori diagnosis after treatment was made by [13C]UBT. The ulcer healing rate was 85.7% in the rebamipide group and 79.5% in the teprenone group (P = NS). The eradication rate was 68.4% (95% CI = 54-83%) in the rebamipide group and 47.7% (95% CI = 32-61%) in the teprenone group (P = 0.043) by per-protocol analysis. These findings suggest that the efficacy of dual therapy may be increased by the administration of rebamipide. PMID- 9753251 TI - Effect of rebamipide on Helicobacter pylori infection in patients with peptic ulcer. AB - This study was designed to assess whether the gastroprotective drug, rebamipide, aids in eradication of H. pylori. One hundred twenty patients, endoscopically diagnosed with gastric or duodenal ulcers and H. pylori infection, were randomly allocated to two treatment groups. Sixty patients received 40 mg of omeprazole twice a day, 1500 mg of amoxicillin three times a day, and 300 mg of rebamipide three times a day (group OAR); the other 60 patients received the same dosage of omeprazole and amoxicillin but no rebamipide for two weeks (group OA). All patients subsequently received an H2-receptor antagonist for six weeks. At the end of the treatment, endoscopy was performed to assess the status of the ulcers as well as the extent of H. pylori infection. In the intent-to-treat (73.3 vs 51.7%, P = 0.014) and per-protocol analyses (75.9 vs 55.3%, P = 0.021) the cure rates for H. pylori infection in group OAR were found to be significantly higher than those in group OA. Our findings suggest that rebamipide aids in curing H. pylori infection. This drug does not induce formation of resistant colonies and has few side effects. PMID- 9753252 TI - Inhibitory effects of rebamipide on ENNG-induced duodenal carcinogenesis in mice: a possible strategy for chemoprevention of gastrointestinal cancers. AB - Rebamipide is a potent antioxidative agent; it increases gastric mucosal PGE2 production and thus protects the gastric mucosa. We hypothesized that the mechanisms of ulcer formation could be extended to carcinogenesis and that an increase in gastric mucosal protection may result in a decrease in gastric carcinogenesis. Therefore, we assessed the inhibitory effects of rebamipide on N ethyl-N'-nitro-N-nitrosoguanidine (ENNG) -induced carcinogenesis in mice. The percentage of tumor-bearing mice in three treatment groups--ENNG + rebamipide 20 mg, ENNG + rebamipide 50 mg, and ENNG alone--was 55%, 42%, and 67%, respectively. The incidence of tumorigenesis tended to decrease with increasing doses of rebamipide. The difference between ENNG + rebamipide 50 mg and ENNG alone was statistically significant (P < 0.05). These results suggest that rebamipide may strengthen the host defense mechanisms related to carcinogenesis in the digestive tract. PMID- 9753253 TI - Absorptive function following small intestinal transplantation. AB - All studies involving small intestinal transplantation and absorptive function are reviewed. The effects ischemia-reperfusion, lymphatic disruption, denervation, rejection, immunosuppressive medication, and infection are elucidated as far as the studies allow. Species differences are discussed. Conclusions regarding the major absorptive defects are drawn. PMID- 9753254 TI - Role of splenectomy in human liver transplantation under modern-day immunosuppression. AB - Between January 1987 and October 1991, 1466 patients underwent consecutive Orthotopic Liver Transplantation (OLTx) at the University of Pittsburgh. Forty of these patient's had concomitant splenectomy with OLTx. These patients were compared to 147 randomly selected OLTx patients without splenectomy within the same time period. One-year patient and graft survival (PS and GS) were lower in splenectomized (Splx) patients compared to nonsplenectomized (non-Splx) patients (59% vs 86% PS, 55% vs 80% GS, respectively). One-month and one-year patient mortality in the Splx group was higher than in the non-splx patients (20% vs 3.4%, P < 0.001 for one month; 40% vs 14.3%, P = 0.003 for one year, respectively). One-month and one-year sepsis-related mortality was also high in Splx patients (17.5% vs 2.7%, P = 0.0022, for one month, and 30% vs 11.5%, P = 0.0043, for one year, respectively). We conclude that concomitant splenectomy with OLTx has a significantly higher patient mortality mainly due to its septic complications and, at present, unless there is a specific indication for a splenectomy, the routine addition of this procedure to liver allograft surgery would not be recommended. PMID- 9753255 TI - Influence of method of alveolar air collection on results of breath tests. AB - The influence of the method alveolar air collection on measurements of trace gas concentration has received little attention. We measured the concentrations of H2, CH4, CO, and CO2 in sequential fractions of alveolar air collected with and without breath-holding. Without breath-holding, the concentration of these gases increased appreciably as increasing quantities of alveolar air were expelled. Twenty seconds of breath-holding markedly reduced this nonhomogeneity of alveolar air. Prediction of the excretion rate of trace gases from measurements of their concentration relative to CO2 and literature values for resting CO2 excretion underestimated the true excretion rate. We conclude that breath-holding prior to sample collection enhances the reproducibility of trace gas measurements. When calculating the rate of excretion of trace gases, the use of literature values for resting ventilation or CO2 excretion may result in appreciable underestimations of the true rate. PMID- 9753256 TI - Assessment of intestinal permeability with a two-hour urine collection. AB - The differential urinary excretion of orally administered lactulose and mannitol is used to evaluate intestinal permeability. This test usually involves a 5- to 6 hr urine collection. We hypothesized that a shorter collection time would give an equivalent result. Forty-three patients with a variety of gastrointestinal symptoms and diagnoses (group 1) and 42 patients with Crohn's disease (group 2) had a standard lactulose/mannitol permeability test. The lactulose and mannitol urinary excretion was calculated using the first urine (group 1) or the 1-hr and 2-hr urine (group 2) and was compared to the values calculated from the routine 5 or 6-hr collection. Lactulose excretion kinetics, expressed as the percent of the total urinary excretion within a given time period, were as follows: 21% in first hour (group 2), 29% in second hour (group 2), and 46% in first 2.5 hr (group 1). Mannitol urinary excretion kinetics were 16%, 31%, and 44%, respectively. The lactulose/mannitol ratio based on a standard urine collection correlated well with the ratio based on just the first urine produced by the patient (R2 = 0.94; P < 0.001; group 1) and the 2-hr urine (R2 = 0.464; P < 0.001; group 2). Future use of the lactulose/mannitol ratio to assess intestinal permeability may be able to be simplified by shortening the urine collection time. PMID- 9753257 TI - Tricyclic antidepressants for functional nausea and vomiting: clinical outcome in 37 patients. AB - Tricyclic antidepressants (TCAs) have been used successfully in the treatment of irritable bowel syndrome and unexplained chest pain. Little information is available regarding their use in other functional gastrointestinal disorders. Clinical charts were analyzed from 37 outpatients (mean age 45 +/- 2 years, 25 females/12 males) with chronic nausea and vomiting that could not be explained by any conventional organic disorder (mean duration of symptoms 28 +/- 8 months). Twenty-one (57%) had chronic persistent symptoms; 16 (43%) had intermittent relapsing symptoms; 13 (35%) also had pain as a dominant complaint. Each patient had been treated with TCAs specifically for the gastrointestinal symptoms (amitriptyline, desipramine, nortriptyline, doxepin, or imipramine), and the subject group was followed for 5.4 +/- 1.1 months. Response (at least moderate symptom reduction using a priori chart rating criteria) occurred in 31 patients (84%), and complete symptom remission occurred in 19 (51%)--in 41% with the first TCA trial. Dose at response averaged 50 mg/day, and outcome was unrelated to TCA used. Logistic regression analysis revealed that pain dominance interfered with remission (P = 0.03), but other clinical characteristics were not predictive of outcome. This uncontrolled clinical experience indicates that the open-label response rate of functional nausea and vomiting to low dosages of TCAs resembles that noted in irritable bowel syndrome. TCAs should be studied in controlled fashion for this and related dyspeptic syndromes, as the success of other treatments is limited. PMID- 9753258 TI - Nonsteroidal antiinflammatory drugs could reverse Helicobacter pylori-induced apoptosis and proliferation in gastric epithelial cells. AB - It remains controversial whether the harmful effects of Helicobacter pylori (Hp) and nonsteroidal antiinflammatory drugs (NSAIDs) are additive. We studied the effects of Hp (virulent and nonvirulent strains) and NSAIDs, alone or in combination, on apoptosis and proliferation of gastric epithelial cells in nonulcer dyspepsia (NUD) patients. Forty-four (25 Hp-positive and 19 Hp-negative) consecutive Chinese NUD patients with rheumatoid arthritis who had taken continuously NSAIDs for more than three months were recruited for this study. Another 41 (20 Hp-positive and 21 Hp-negative) NUD patients not on any NSAIDs were included as controls. All patients underwent a gastroscopy examination and gastric biopsies. Hp infection was confirmed by CLOtest, anti-Hp ELISA, and [13C]urea breath test. The CagA status was determined by the anti-CagA antibody assay. The degree of gastritis, apoptosis, and proliferation indices were determined with H&E staining, terminal uridine deoxynucleotidyl nick end-labeling (TUNEL), and proliferating cell nuclear antigen (PCNA) immunostaining methods, respectively. A significantly higher apoptosis was observed in subjects who had Hp infection or had been consuming NSAIDs when compared with the controls. Unlike NSAID-treated subjects, patients with Hp infection were shown to have significantly enhanced cell proliferation. However, the increased apoptosis and proliferation in Hp-positive subjects were reversed by also taking NSAIDs. No correlation was found between apoptosis and proliferation in all the study groups. There was no association found between CagA expression or degree of gastritis with cell proliferation or apoptosis. It was demonstrated at the cellular level that NSAIDs could abrogate apoptosis or proliferation effects induced by Hp. Furthermore, the latter effects appeared not to be influenced by the virulent nature of the Hp strains. PMID- 9753260 TI - "Deathgrip" esophagus: segmental aperistalsis following electrical injury. AB - We report a case of documented transient segmental aperistalsis of the distal esophagus following an accidental electrical injury. There are no other reports in the literature demonstrating this phenomenon. A review of gastrointestinal injury secondary to electrical injury is presented. PMID- 9753259 TI - Omeprazole and ranitidine in long-term treatment of duodenal ulcer: a double blind comparison of length of time in remission. AB - In most patients duodenal ulcer is a chronic relapsing disease. If no active maintenance treatment or eradication therapy is given after healing, around 70 100% of patients have a relapse during the first year. We conducted a double blind multicenter study in 472 patients with duodenal ulcer. They were treated with omeprazole 20 mg every morning for four or eight weeks and when healed were randomly allocated to maintenance treatment with either omeprazole 20 mg every morning or ranitidine 150 mg at bedtime for up to six months. The patients were assessed by endoscopy at monthly intervals until healing occurred. Thereafter scheduled endoscopy was carried out after 1, 3, and 6 months of maintenance treatment or immediately in the event of a suspected relapse. Healing status (intention to treat approach) was 87% at four weeks and 93% at eight weeks. At six months the estimated remission rate was 90% for omeprazole and 82% for ranitidine (P = 0.03, 95% CI 1-15%). The incidence of adverse events was similar during the two maintenance treatments. Treatment with omeprazole 20 mg every morning maintained significantly more patients in remission than treatment with ranitidine 150 mg at bedtime. PMID- 9753261 TI - Pneumatic dilatation is effective long-term treatment for achalasia. AB - Although pneumatic dilatation (PD) has been an established treatment for achalasia for decades, there is limited information on its long-term clinical efficacy. We have followed up the clinical status of patients having PD with a 30 or 35-mm balloon by one of us (D.O.C.) over a 25-year period. Of 144 patients whose initial records were available for review, 31 could not be contacted. Of the remaining 113 patients, 72 (64%) responded to a questionnaire assessing swallowing status and patient satisfaction, and this forms the basis of this report. There were 32 men and 40 women, with mean age 46 years (range: 17-78); mean length of follow-up since PD was 6.5 years (range: 10 months to 25 years). Success was primarily defined by the need for no additional therapy for achalasia other than one or two PD's. PD was effective long-term treatment in 61/72 patients (85%); only four of these required a second PD over this time interval. There was no significant difference in any of the following parameters between patients with a treatment success or failure: age, sex, size of pneumatic dilator, and duration of symptoms prior to PD. Response was significantly better (P < 0.05) in patients having no prior dilatation (43/47; 91%) than in those in whom another physician had performed prior dilatation (18/25; 72%). In response to the question of whether they would select PD again, 68 patients (94%) responded positively. In conclusion, pneumatic dilatation performed using a consistent technique by an experienced physician is effective long-term therapy for achalasia patients of all ages. Most patients require only one dilatation. PMID- 9753262 TI - Intersubject and interswallow variability in topography of esophageal motility. AB - Topographic plots linking averaged manometric data in time and space suggest that sequential contraction segments form esophageal peristalsis. A system capable of plotting individual swallows was developed to verify this observation and to determine intersubject and interswallow variability in their topographic appearance. Fourteen volunteers were studied with a novel computerized assimilation method capable of generating topographic contour plots as well as conventional wave forms for analysis. Contraction segments in the proximal body and lower sphincter were identified in all subjects as being separate from the remainder of the esophagus with little interswallow variation. The appearance of peristalsis through the distal body was more variable because of its intermittent separation into two dominant contraction segments (59.8% of swallows) that had poorly correlated contraction strength (median r = 0.15). Intersubject variability exceeded interswallow variability in topographic landmarks, resulting in distinctive topographic "fingerprints" of peristalsis for each subject. We conclude that topographic plotting of single swallows is feasible and confirms the presence of sequential contraction segments in the esophagus. Interswallow variability helps demonstrate two segments within the smooth-muscle body, an anatomical region of seeming homogeneity, that have sufficient contraction independence to indicate separate neuromuscular units responding to different contractile influences. PMID- 9753264 TI - Gastroesophageal reflux disease is a common cause of noncardiac chest pain in a country with a low prevalence of reflux esophagitis. AB - Gastroesophageal reflux disease is believed to be uncommon in the East. This study aimed to determine if such a condition was a significant cause of noncardiac chest pain in Singapore. Eighty consecutive patients with recurrent chest pain, who had cardiac and other obvious causes excluded, underwent esophagogastroduodenoscopy, standard manometry, acid perfusion test, and prolonged ambulatory pH and pressure monitoring. Endoscopic esophagitis, positive acid perfusion tests, pathologic reflux, and positive chest pain-reflux correlation were detected in 7/80 (8.8%), 11/70 (15.7%), 14/61 (23.0%), and 12/25 (48.0%) patients, respectively. Among those with pathologic reflux, endoscopic esophagitis was present in only two (14.3%). Overall, 32 (40%) patients had gastroesophageal reflux disease. Esophageal motility disorder, alone or in association with gastroesophageal reflux disease, was demonstrated in only five (6.3%) patients. Our results confirmed western reports that gastroesophageal reflux disease was a common cause of noncardiac chest pain, whereas motility disorder was an infrequent cause of such pain. PMID- 9753263 TI - Cisapride enhances the effect of partial posterior fundoplication on esophageal peristalsis in GERD patients with poor esophageal contractility. AB - Partial posterior fundoplication improves esophageal peristalsis in patients with gastroesophageal reflux disease (GERD) associated with poor esophageal body function. The aim of this study was to investigate whether postoperative administration of cisapride enhances the effect of surgery on esophageal peristalsis. Laparoscopic partial posterior fundoplication was performed on 34 consecutive GERD patients with poor esophageal body motility. These patients were randomized in groups without and with postoperative treatment with cisapride 20 mg twice daily for six months. Esophageal manometry was performed preoperatively and six months following surgery. Esophageal body function improved significantly following partial posterior fundoplication without or with postoperative treatment with cisapride. However, this effect was more pronounced in the group of patients receiving cisapride. Partial posterior fundoplication combined with postoperative treatment with cisapride should be the therapy of choice in GERD patients with poor esophageal body motility. PMID- 9753265 TI - Esophageal irritation due to alendronate sodium tablets: possible mechanisms. AB - Animal studies were done using an in vivo dog model to examine the possible mechanism for the esophageal adverse events reported with alendronate sodium tablets. These studies showed that under low pH conditions alendronate sodium can cause esophageal irritation. No esophageal irritation occurred at pH 3.5 or higher where the drug exists primarily as the sodium salt. The animal studies also showed that alendronate sodium can exacerbate preexisting esophageal damage. Exposure of the esophageal mucosa for a prolonged period to alendronate sodium tablet can also cause mild esophageal irritation. These findings suggest that the esophageal irritation in patients taking Fosamax can be from prolonged contact with the tablet, reflux of acidic gastric contents with alendronate sodium, and exacerbation of preexisting esophageal damage. The findings also suggest that other bisphosphonates can cause esophageal injury under similar conditions. PMID- 9753266 TI - Effects of cyclooxygenase-2 selective and nitric oxide-releasing nonsteroidal antiinflammatory drugs on mucosal ulcerogenic and healing responses of the stomach. AB - Effects of selective cyclooxygenase-2 (COX-2) inhibitors (NS-398) and nitric oxide (NO) -releasing aspirin (NO-ASA) on gastric ulcerogenic and healing responses were examined in comparison with nonselective COX inhibitors such as indomethacin and aspirin (ASA). Hypothermic stress (28-30 degrees C, 4 hr) induced gastric lesions in anesthetized rats with an increase of acid secretion. The lesions induced by hypothermic stress were markedly worsened by subcutaneous administration of both indomethacin and ASA but were not affected by either NS 398 or NO-ASA, although the increased acid secretion during hypothermia was not affected by any of the drugs. On the other hand, the healing of gastric ulcers induced in mice by thermal cauterization (70 degrees C, 15 sec) was significantly delayed by daily subcutaneous administration of indomethacin and ASA as well as NS-398, but not by NO-ASA. COX-2 mRNA was not detected in the intact mucosa but was positively expressed in the ulcerated mucosa, most potently on day 3 after ulceration. Prostaglandin contents in the intact mouse stomach were reduced by indomethacin, ASA, and NO-ASA, while the increased prostaglandin generation in the ulcerated mucosa was inhibited by all drugs including NS-398. After subcutaneous administration of NO-ASA to pylorus-ligated rats and mice, high amounts of NOx were detected in both the gastric contents and serum. In addition, both NS-398 and NO-ASA showed an equipotent antiinflammatory effect against carrageenan-induced paw edema in rats as compared with indomethacin and ASA. These results suggest that both indomethacin and ASA not only increased the mucosal ulcerogenic response to stress but impaired the healing response of gastric ulcers as well. The former action was due to inhibition of COX-1, while the latter effect was accounted for by inhibition of COX-2 and was mimicked by the COX-2-selective inhibitor NS-398. NO-ASA, although it inhibited both COX-1 and COX-2 activity, had no deleterious effects on gastric ulcerogenic and healing responses. PMID- 9753268 TI - Gastric inflammation during systemic anaphylaxis: neutrophil recruitment in stomach wall of mice does not require mast cell participation. AB - We determined whether neutrophil infiltration into the stomach wall occurred during systemic anaphylaxis in mice and assessed the participation of mast cells in the response. Normal mice sensitized and challenged with antigen exhibited significant neutrophil infiltration in the gastric mucosa and submucosa compared with saline-challenged mice. The development of clinical signs of anaphylaxis and extent of gastric neutrophil infiltration was similar in mast cell-deficient Kit(W)/Kit(W-v) or Mgf(Sl)/Mgf(Sl-d) mice and the respective normal congenic mice. Pretreatment with sodium cromoglycate prevented the clinical signs of anaphylaxis and significantly diminished the infiltration of neutrophils in +/+ or Kit(W)/Kit(W-v) mice. Systemic anaphylaxis is associated with neutrophil infiltration into the stomach wall in mice, and mast cells are not required for the development of either the clinical manifestations or gastric neutrophil infiltration observed in the response. PMID- 9753267 TI - Effect of gastric secretion on penetration of N-3H-methyl-N-nitro-N nitrosoguanidine into gastric mucosa of rats. AB - Clinical conditions with low gastric acid secretion have been associated with increased risk of gastric cancer. There has also been concern about gastric acid inhibition and N-nitroso compound formation in the stomach. This study investigates the effect of gastric acid secretion on the penetration of N-3H methyl-N-nitro-N-nitrosoguanidine, an N-nitroso compound and gastric carcinogen, into the gastric mucosa of rats. Gastric acid secretion was stimulated by pentagastrin (40 microg/kg/hr) and inhibited by omeprazole (40 micromol/kg) before mucosal exposure to N-3H-methyl-N-nitro-N-nitrosoguanidine. Penetration of the carcinogen was evaluated by light microscopic identification of cells in the S-phase labeled with N-3H-methyl-N-nitro-N-nitrosoguanidine. This population of double-labeled cells is considered at risk from N-methyl-N-nitro-N nitrosoguanidine-induced carcinogenesis. The percentage of double-labeled cells was significantly higher in antrum than in corpus mucosa (P < 0.0001). Stimulation or inhibition of gastric acid secretion did not affect the penetration of N-3H-methyl-N-nitro-N-nitrosoguanidine in antrum or corpus mucosa. We conclude that modulation of gastric acid secretion does not affect the penetration of the carcinogen into the gastric mucosa nor does it explain the different penetration of the carcinogen into corpus and antrum mucosa. PMID- 9753269 TI - Disturbed solid-phase gastric emptying in functional dyspepsia: a meta-analysis. AB - Functional dyspepsia is a common disorder with a diverse pathophysiological background, but the role of motility disorders in functional dyspepsia remains unclear. We aimed to quantify the relationship between disturbed gastric emptying and functional dyspepsia, using a meta-analytic approach. Through a structured literature search of Medline and Embase from 1983 to 1996, we selected all studies in which scintigraphic solid-phase gastric emptying was measured in both functional dyspeptic patients and controls. Seventeen studies involving 868 dyspeptic patients and 397 controls were pooled. Gastric emptying in patients with functional dyspepsia was 1.46 (1.23-1.69) times slower than controls; the proportion of patients with abnormally slow emptying was either 37% (34-40%, simple numeric pooling) or 39% (29-49%, weighted pooling). We conclude that gastric emptying of solids in patients with functional dyspepsia is 1.5 times slower than in healthy controls and that a significant delay of emptying is present in almost 40% of patients with functional dyspepsia. PMID- 9753270 TI - Mechanical properties and collagen content differ between isolated guinea pig duodenum, jejunum, and distal ileum. AB - We compared the stress-strain distributions obtained from isolated segments of the guinea pig duodenum, jejunum, and distal ileum, and the relation between the elastic properties and the collagen content. The segments were immersed in Krebs Ringer solution containing 10(-2) M MgCl2 to abolish contractile activity. Stepwise inflation of an intraluminal balloon in which the cross-sectional area (CSA) was measured provided the luminal pressure-loading stimulus. The wall thickness was measured by means of 20-MHz A-mode ultrasound. The stress-strain and the incremental elastic modulus-strain distributions were derived from the steady-state values of wall thickness, internal radius, and applied pressure. The CSA-pressure relations and the wall thickness-pressure relations were nonlinear and both differed between the segments (P < 0.01). The stress-strain distributions showed an exponential behavior that fitted well to the equation Y = a x Exp(b x X) (r2 = 0.97 +/- 0.01). The intercept with they axis (a) and the slope of the curves (b) differed between the segments (P < 0.01 and P < 0.05). The collagen contents were 3.99 +/- 0.18 microg/mg, 2.51 +/- 0.13 microg/mg, and 2.10 +/- 0.11 microg/mg in the duodenum, jejunum, and distal ileum, respectively. This difference was significant among all three locations (P > 0.05). An association was found between the collagen content and the incremental elastic modulus (stiffness) at a stress level of 70 kPa (P < 0.05). PMID- 9753271 TI - Cooperative roles of colon and anorectum during spontaneous defecation in conscious dogs. AB - Colorectal motility during spontaneous defecation was investigated using force strain gauge transducers implanted in the proximal colon, distal colon, rectum, and anus in six dogs. One 24-hr recording and several defecation recordings were made in each dog. During 24-hr recordings, 29 giant contractions were observed in the distal colon. The giant contractions, which propagated to the rectum, accompanied evacuation more frequently than those that stopped at the distal colon (P < 0.05). Of 66 episodes of defecation, 63 (95%) were accompanied by a giant contraction of the distal colon. Of these, 57 (90%) propagated to the rectum. In three events, giant contraction originated at the rectum. The rectum relaxed prior to the contraction. The internal anal sphincter also relaxed. Migration of giant contraction to the rectum, rectal relaxation-contraction sequence and sphincter relaxation played important roles during defecation. Defecation is a consequence of successive phenomena occurring in both the colon and anorectum. PMID- 9753272 TI - Decrease in gastric sensitivity to distension by 5-HT1A receptor agonists in rats. AB - Using an in vivo model for evaluation of gastric sensitivity in awake rats, we aimed to determine whether 5-hydroxytryptamine 1A (5-HT1A) agonists modify pain threshold and gastric compliance specifically through 5-HT1A receptors. Isobaric gastric distensions were performed with a barostat using steps of 5 mm Hg in male rats equipped with a gastric balloon and electrodes implanted in the neck muscles. Gastric distension at 15 or 20 mm Hg induced a typical posture associated with contractions of the neck muscles. Rats received drugs 30 min before gastric distension. The 5-HT1A receptor agonist 8-hydroxy-2-(di-n propylamino)tetralin (8-OH-DPAT), administered intraperitoneally (0.5 mg/kg) increased gastric pain threshold and gastric tone. These effects were reproduced when administered centrally (0.05 mg/kg) and blocked by intracerebroventricular administration of the 5-HT1A antagonist WAY 100635. Flesinoxan (4 mg/kg, intraperitoneally), another 5-HT1A agonist reproduced the effects of 8-OH-DPAT on pain threshold and gastric tone and the alpha2-receptor antagonist yohimbine did not modify the action of 8-OH-DPAT. Our results indicate that activation of 5 HT1A receptors at the level of the central nervous system increases gastric tone and decreases gastric sensitivity to distension. PMID- 9753273 TI - Brewer's yeast and Saccharomyces boulardii both attenuate Clostridium difficile induced colonic secretion in the rat. AB - Saccharomyces boulardii (Sb), a nonpathogenic yeast, has been used to prevent recurrences of Clostridium difficile (C.diff) -associated diarrhea. A single report suggested that treatment with Saccharomyces cerevisiae (Sc), commonly called brewer's yeast (BY), facilitates treatment of persistent C.diff infection. We conducted this experiment to determine whether C.diff toxin A-induced colonic secretion in the rat is blunted by pretreatment with either Sb or BY. We employed closed cecal pouches in two groups of five adult male Sprague-Dawley rats fed with standard chow for five days prior to the experiment, another group whose water was supplemented with 20 x 10(9) colony-forming units (CFU) of Sb per day for five days, and another group whose water was supplemented with 20 x 10(9) CFU of Sc per day for five days. Cecal pouches were infused for 3 hr with one of the following: (1) normal saline alone for a control group, or (2) normal saline plus 5 microg of C.diff toxin A (for the other control group and for the two experimental groups). Water movement was measured by a nonabsorbable marker technique. Sodium movement and permeability to mannitol were also measured. Prior to the infusion, cecal contents were quantitatively cultured. In the three animals whose ceca were colonized with less than 10(6) CFU of either yeast per gram wet cecal content, toxin A-induced secretion could not be attenuated. In contrast, animals whose ceca were colonized with more than 10(6) CFU of either yeast per gram of wet cecal content showed significantly less secretion after toxin A application than those which were not fed yeast. S. cerevisiae reduced secretion by half (N = 5, P = 0.039 for water, 0.044 for sodium) and Sb by 75% (N = 4, P = 0.015 for water, 0.034 for sodium). Toxin-induced increases in permeability to [3H]mannitol from systemic circulation to cecum could not be blunted by either yeast. We conclude that rat ceca can be colonized by either organism and that both organisms reduce C.diff toxin A-mediated secretion. We speculate that both organisms might have benefit in human C.diff-associated enterocolitis. Further studies of their mechanisms of action as well as clinical trials for the prevention and treatment of human C.diff infections should be pursued. PMID- 9753274 TI - Recombinant or plasma-derived antisecretory factor inhibits cholera toxin-induced increase in Evans blue permeation of rat intestinal capillaries. AB - The effect of cholera toxin on small intestinal capillary function, utilizing the Evans blue dye method, was analyzed. The modulatory influence of plasma-derived or recombinant human antisecretory factor on this variable was also investigated. Male Sprague-Dawley rats were briefly anesthetized with ether, and a jejunal loop was constructed that was challenged for 90 min with phosphate-buffered saline or cholera toxin. Five minutes prior to death, the rats received an intravenous injection of Evans blue. The tissue content of dye in the loop was quantitated spectrophotometrically or demonstrated histochemically. Cholera toxin increased the recovery of Evans blue; extravasation of the dye was prominent in the top of the villi, while the crypts were spared. It is suggested that the toxin caused increased transcapillary permeation of albumin in a heterogenous fashion in the gut wall. This effect of the toxin was prevented by pretreatment with the antisecretory factor. PMID- 9753275 TI - Mucosal expression of interleukin-6 and interleukin-8 messenger RNA in ulcerative colitis and in Crohn's disease. AB - To elucidate the possible role of proinflammatory cytokines in inflammatory bowel disease, the expression and localization of interleukin (IL) -6 and IL-8 mRNAs were examined in colonic biopsy specimens obtained from 10 patients with active ulcerative colitis (UC), 5 with inactive UC, 6 with Crohn's disease (CD), and 5 normal controls. In situ hybridization with digoxigenin-labeled probes and immunohistochemistry for both cytokines were performed. The IL-6 mRNA expression was enhanced in the inflamed mucosa in 4 of 6 CD patients, while that of UC patients stayed at baseline. In contrast, IL-8 mRNA expression was apparently augmented (P = 0.044) in 7 of 10 active UC and 3 of 6 CD patients (NS). The cell count positive for IL-8 mRNA per unit area was definitely increased in moderate/severe UC when compared to mild UC (53.1 +/- 14.4/mm2 vs 9.0 +/- 5.1/mm2, P = 0.028) according to the degree of inflammation. IL-6 mRNA positive cells in CD were preferentially located in deeper lamina propria than IL-8 mRNA positive cells in UC. Interestingly, IL-8 mRNA was expressed in the mucosal epithelial cells in one UC patient. The patients treated by corticosteroids tended to show suppressed expression of each mRNA, except one patient with intractable UC. Our data suggest enhanced expression of mucosal IL-6 mRNA in CD and of IL-8 mRNA in UC by infiltrating mononuclear cells, indicating the distinct participation of each cytokine in the pathogenesis of UC and CD. Moreover, intestinal epithelial cells in UC occasionally exhibit IL-8 mRNA. PMID- 9753277 TI - Carbonic anhydrase I reduction in colonic mucosa of patients with active ulcerative colitis. AB - Ulcerative colitis (UC) is associated with low intracolonic pH and unbalanced transmucosal ionic exchanges. Along the gastrointestinal tract carbonic anhydrase isoenzyme I (CA-I) is specifically expressed in colon epithelium and is involved in mucosal control of ion, fluid, and acid-base balance. Since altered CA-I expression may play some role in UC, CA-I was measured at the mRNA and protein level and carbonic anhydrase (CA) enzyme activity was determined in colon biopsies of 14 UC patients (6 remission, 4 mild, 4 moderate UC) and of 12 healthy subjects. Patients with mild or moderate UC showed a significant reduction of CA I mRNA and protein and of total CA activity in the inflamed mucosa compared to controls. Patients with UC in remission showed a pattern of CA-I expression and CA activity similar to controls. This is the first report showing a reduction in the expression of CA-I in active UC. PMID- 9753278 TI - Evidence for autonomic nervous system imbalance in women with irritable bowel syndrome. AB - Autonomic nervous system function was assessed in women with and without irritable bowel syndrome using frequency domain measures of heart rate variability. Women were interviewed and placed into the irritable bowel syndrome (N = 25) group based on history of diagnosis and self-report of current gastrointestinal symptoms. Women in the control group denied a history of chronic gastrointestinal symptoms (N = 15). Women were followed for one menstrual cycle with a symptom diary, and during mid-luteal phase they wore a Holter 24-hr electrocardiograph monitor. Women with irritable bowel syndrome demonstrated significantly lower vagal tone as measured by the high frequency spectrum relative to control women. In addition, women with irritable bowel syndrome had a flattened 24-hr pattern of heart rate variability, with significantly lower levels of vagal tone during sleep. These results suggest that systemic sympathovagal balance may be shifted in a subset of women with irritable bowel syndrome. PMID- 9753276 TI - Sulfur metabolism in ulcerative colitis: investigation of detoxification enzymes in peripheral blood. AB - Two enzymes of detoxification were studied in blood samples from 27 patients with ulcerative colitis (UC) and 18 controls to determine whether there is an abnormality in sulfur metabolism in UC. Thiol methyltransferase (TMT) activity was measured in erythrocyte membranes as the extent of conversion of 2 mercaptoethanol to S-methyl-2-mercaptoethanol with [3H]methyl-S-adenosyl methionine as methyl donor. Phenol sulfotransferase (PST) activity was measured in platelet homogenates as the extent of sulfation of p-nitrophenol with 3 phosphoadenosine 5-phospho[35S]sulfate (PAPS) as sulfate donor. TMT activity was significantly higher in UC patients (27.0 vs 17.1 nmol/mg protein/hr; P < 0.005). No difference in PST activity was found. We conclude that TMT may be up-regulated in UC to detoxify excess hydrogen sulfide exposed to the peripheral blood compartment. This may arise from either increased luminal sulfide production or reduced colonic detoxification. PMID- 9753280 TI - Quantitative and ultrastructural analysis of rectal mucosal mast cells in acute infectious diarrhea. AB - The role of mast cells, potential mediators of mucosal immunity and inflammation, was studied morphologically in the rectal mucosa in two acute diarrheal diseases, cholera and shigellosis. Quantitation of mucosal mast cells showed that they were significantly higher in the deeper lamina propria where blood vessels and nerves were more abundant. There was no difference in mast cell counts or degranulation in the mucosa in both groups of patients and controls. Intraepithelial mast cells were decreased in the patients. The prevalence of lipid bodies was significantly higher in mast cells from patients with cholera and shigellosis (P < 0.01). These findings suggest that mast cell populations are more dense around blood vessels and nerves and that inflammatory mediators derived from arachidonic acid metabolites, as indicated by the lipid bodies, are the response of mast cells to the alterations in diarrhea, despite differences in the etiology of diarrhea. PMID- 9753279 TI - Intestinal absorption of nutrients is not influenced by soy fiber and does not differ between oligomeric and polymeric enteral diets. AB - Enteric feeding is often associated with diarrhea. To avoid this side effect, isoosmotic and fiber-supplemented enteral diets are recommended. The aims of this study were to determine whether supplementing enteral diets with soy fiber influences nutrient absorption and whether in enteric feeding absorption of nutrients and water fluxes differ between hyperosmotic oligomeric and isoosmotic polymeric diets. In mini pigs intestinal absorption and water fluxes were measured by perfusing a 150-cm length of jejunum. Six noncommercial iso- and hyperosmotic oligomeric and polymeric diets and six commercial polymeric diets, either fiber-free or supplemented with soy fiber, were used. Pancreatic enzymes were infused concomitantly with the polymeric diets. The absorption of nutrients and energy did not differ between oligomeric and polymeric diets. Oligomeric diets of high energy density produced a pronounced secretion of water. Despite lower initial osmolality, polymeric diets produced a similar secretion of water due to rapid pancreatic hydrolysis. Supplementing diets with largely insoluble soy fiber increased viscosity only between 4.6 and 14.5 mPa x sec. Soy fiber did not influence absorption of nutrients and energy and had also no effects on luminal transit and flow rate. The lack of effects was not due to dilution of chyme by intestinal secretion of water because no differences existed between isoosmotic and hyperosmotic oligomeric diets. In conclusion, supplementing enteral diets with soy fiber does not impair the absorption of nutrients. Enteric feeding with isoosmotic polymeric diets provides no advantage compared with hyperosmotic oligomeric diets with respect to absorption of nutrients and secretion of water. PMID- 9753281 TI - Effect of preinduction of heat-shock proteins on acetic acid-induced small intestinal lesions in rats. AB - Bowel dysfunction such as irritable bowel syndrome caused by stress is well described. Previous reports suggest that stress is known to cause the release of endogenous substances such as catecholamine, beta-endorphine, 5 hydroxytryptamine, corticotropin-releasing factor, and thyrotropin-releasing hormone (TRH). However, the role played by these neurohormonal mediators in bowel dysfunction under stress conditions is not well known. We investigated the influence of water-immersion stress or TRH administration on the expression of 60 kDa, 72-kDa, and 90-kDa heat-shock proteins (HSP60, HSP72, and HSP90, respectively) in rat small intestinal mucosa by Western blot and immunohistochemical analyses. The cytoprotective function of preinduced HSPs on experimentally induced mucosal damage also was studied. In order to investigate the influence of preinduction of HSP60 on small intestinal damage, the small intestinal lumen was perfused with 1.5% acetic acid 1 ml/min for 15 min with or without pretreatment with water-immersion stress or TRH administration. Expression of HSP60 was significantly increased by water-immersion stress or TRH administration in the small intestinal mucosa, whereas HSP72 and HSP90 did not increase. Interestingly, expression of this protein showed the biphasic peak pattern after water-immersion stress or TRH administration. Each peak was observed 3-6 hr and 21-24 hr after the initiation of water-immersion stress or TRH administration. Immunohistochemical study also showed a significant increment of HSP60 in both the cytoplasm and nuclei of the small intestinal mucosal cells. No histopathologic alteration was observed in rat small intestinal mucosa after each treatment. Small intestinal damage caused by 1.5% acetic acid perfusion was not influenced by preinduction of HSP60. We demonstrated that water-immersion stress or TRH administration specifically induced HSP60, although preinduction of this protein did not show a cytoprotective function in the small intestinal mucosa. PMID- 9753282 TI - Cholelithiasis and dietary risk factors: an epidemiologic investigation in Vidauban, Southeast France. General Practitioner's Group of Vidauban. AB - Dietary risk factors have been implicated in the development of cholelithiasis. The aim of this study was to determine in a homogeneous French population whether a particular type of diet may be lithogenic. Seventy-six subjects over 30 years of age (26 men, 50 women) with cholelithiasis detected by ultrasound were selected from a population sample of 830 subjects by drawing lots using the polling list. These were matched by 76 control subjects without cholelithiasis randomly selected from the same population. Univariate analysis was significant for a high calorie diet >2500 kcal/day (OR = 3.62, P = 0.0065), a diet rich in carbohydrates with a consumption > or = 55 g/day (OR = 2.98, P = 0.0067), and a diet rich in total lipids (OR = 4.97, P = 0.023) or saturated fatty acids (OR = 3.06, P = 0.0146). An alcohol consumption equivalent to 20-40 g/day was protective (P = 0.018). Multivariate analysis confirmed these results. Our study suggests that a change in dietary habits by limiting excess calories, saturated fats and carbohydrates could reduce the incidence of cholelithiasis. PMID- 9753283 TI - Primary biliary cirrhosis associated with cholangiocarcinoma. PMID- 9753284 TI - Hepatitis G virus in the general population and in patients on hemodialysis. AB - To determine the routes of transmission of hepatitis G virus (HGV) and the relationship between HGV and hepatitis C virus (HCV) infections, we tested for HGV RNA by polymerase chain reaction and antibody to HCV (anti-HCV) in 494 hemodialysis patients, 638 inhabitants of two HCV endemic areas, and in 400 blood donors in Japan. HGV RNA was detected in 6.9% of hemodialysis patients, in 1.4% of inhabitants, and in 0.8% of donors, and anti-HCV was detected in 39.3%, 12.4%, and 1.8%, respectively. Of HGV RNA-positive hemodialysis patients, and HGV RNA positive inhabitants, 64.7% and 11.1%, respectively, had been given blood transfusions. The prevalences of HGV RNA and anti-HCV significantly increased with the duration of hemodialysis. Of all HGV RNA positives, 74.4% were coinfected with HCV and subjects with HGV RNA alone had normal liver function. In conclusion, HGV is transmitted by blood transfusion and within the hemodialysis unit itself. HGV does not seem to injure hepatocytes. PMID- 9753286 TI - AST/ALT ratio > or = 1 is not diagnostic of cirrhosis in patients with chronic hepatitis C. AB - Medical guidelines for interferon-alpha2a or -alpha2b (IFN-alpha) treatment of chronic hepatitis C virus (HCV) infection depend upon baseline liver histology. A better long-term response to IFN-alpha therapy correlates with less inflammation and absence of cirrhosis. It has been suggested that the presence of cirrhosis in patients with chronic hepatitis C virus infection may be predicted based on an AST/ALT ratio > or = 1. This study was designed to determine if the presence of cirrhosis can be predicted in patients with chronic HCV infection by such a ratio. Seventy-seven patients, including 23 cirrhotics, with chronic HCV infection were studied. Serum ALT, AST, and HCV-RNA levels and hepatic activity index (HAI), reflecting histologic inflammation in all liver biopsies, were assessed. AST/ALT ratios and mean ALT, AST, and HCV-RNA were determined for both cirrhotic and noncirrhotic patients. HAI was correlated with ALT, AST, and HCV RNA levels, the latter determined by quantitative RT-PCR. The likelihood ratio (LR) and positive predictive value of an AST/ALT ratio > or = 1 for cirrhosis was 7.3 and only 77%, respectively. In cirrhotics vs noncirrhotics, there were no significant differences between mean serum ALT (149 +/- 28 vs 176 +/- 17 units/liter), AST (139 +/- 28 vs 102 +/- 8 units/liter), or HCV-RNA levels (589,160 +/- 147,053 vs 543,915 +/- 75,497 copies/ml), respectively. There was a significant, but clinically weak, correlation between serum ALT and HAI (r = 0.234), and none between HAI and either serum AST or HCV-RNA levels. Our results support the need for a liver biopsy prior to treatment of chronic HCV infection, since the AST/ALT ratio fails to predict accurately the presence of cirrhosis. PMID- 9753287 TI - Treatment of autoimmune cholangitis. PMID- 9753285 TI - Immune phenotype of chronic liver disease. AB - Immune disorders in chronic liver disease may reflect common host propensities or disease-specific factors. Our aim was to determine the principal bases for these expressions. Four hundred fifty-one patients with various chronic liver diseases were assessed prospectively for concurrent immune disorders. Individuals with immune diseases were more frequently women (73% vs 60%, P = 0.02) and they had HLA DR4 more often than counterparts with other HLA (46% vs 23%, P = 0.000008). The association between HLA DR4 and immune disease was apparent within individual liver diseases and within different categories of liver disease. Women with HLA DR4 had a higher frequency of immune disease than women without HLA DR4 (52% vs 22%, P < or = 0.000001), and they also had immune diseases more commonly than DR4 positive men (52% vs 31%, P = 0.03). DR4-positive men, however, had higher frequencies of immune disease than DR4-negative men, especially in the nonimmune types of liver disease (26% vs 4%, P = 0.002). We conclude that HLA DR4 and female gender constitute an immune phenotype that is an important basis for autoimmune expression in chronic liver disease. PMID- 9753288 TI - Granulomatous liver disease and giant-cell arteritis. PMID- 9753289 TI - Apoptosis and the pathogenesis of IDDM: a question of life and death. AB - In type 1 diabetes, an immune-mediated process leads to the destruction of pancreatic beta-cells. In the last decade, considerable progress has been made in understanding the cellular and biochemical pathogenic processes of the disease. However, more needs to be learned about the immune mechanisms leading to the development of autoreactive immune cells and the molecular mechanisms of beta cell death. The study of apoptosis of autoreactive lymphocytes as well as apoptosis of beta-cells may give answers to many still unsolved questions. This review focuses on the possible role of apoptosis both in the regulation of immune mechanisms involved in the pathogenesis of type 1 diabetes and as a way for beta cells to die. The advancement in the knowledge of the possible role of apoptosis and its regulation in the pathogenesis of type 1 diabetes may provide new therapeutic tools for the prevention of the disease. PMID- 9753290 TI - Distinguishing the type I and type II isozymes of hexokinase: the need for a reexamination of past practice. AB - The type I and type II isozymes of hexokinase coexist in insulin-sensitive tissues, such as cardiac and skeletal muscle and adipose tissue. Based on an early report that the purified type I isozyme was stable at 45 degrees C whereas the purified type II isozyme was not, investigators in a number of studies have used heat lability as a criterion for distinguishing these isozymes in crude tissue extracts or subcellular fractions; that is, activity lost after incubation at 45 degrees C was believed to be type II while remaining activity was considered type I. This extrapolation is dangerous because thermal lability can be markedly affected by the solvent environment, including the presence or absence of other proteins. In the present study, the rate of thermal inactivation of the type I isozyme has been shown to vary by at least an order of magnitude in soluble and particulate fractions prepared from rat heart and brain. Thus, the use of thermal stability as a general criterion for identifying the type I isozyme is invalid, and conclusions based on thermal inactivation as a means for distinguishing the type I and type II isozymes need to be reconsidered. PMID- 9753291 TI - Epinephrine and insulin stimulate different mitogen-activated protein kinase signaling pathways in rat skeletal muscle. AB - Little is known about the regulation of the mitogen-activated protein (MAP) kinase signaling cascades by hormonal stimulation in vivo. The extracellular signal-regulated kinase (ERK) and the c-jun kinase (JNK) are two MAP kinase signaling pathways that could play a role in the cellular response to hormones such as insulin and epinephrine. We studied the effects of insulin (20 U/rat) and epinephrine (25 microg/100 g body wt) injected in vivo on ERK and JNK signaling in skeletal muscle from Sprague-Dawley rats. Insulin significantly increased ERK phosphorylation and the activity of its downstream substrate, the p90 ribosomal S6 kinase 2 (RSK2), by 1.4-fold, but it had no effect on JNK activity. In contrast, epinephrine had no effect on ERK phosphorylation or RSK2 activity, but it increased JNK activity by twofold, an effect that was inhibited by the presence of combined alpha and beta blockade. Furthermore, the phosphorylation of both p46 and p55 isoforms of JNK, measured by phosphospecific antibody, was increased severalfold. The activity and phosphorylation of MAP kinase kinase (MKK)-4, an upstream regulator of JNK, was unchanged by epinephrine. Incubation of isolated soleus muscles in vitro with epinephrine (10(-5) mol/l) also increased JNK activity by twofold. These data are the first to demonstrate that epinephrine can increase JNK activity. Insulin and epinephrine have different effects on MAP kinase signaling pathways in skeletal muscle, which may be one of the underlying molecular mechanisms through which these hormones regulate opposing metabolic functions. PMID- 9753292 TI - Increase in insulin action and fat oxidation after treatment with CL 316,243, a highly selective beta3-adrenoceptor agonist in humans. AB - Stimulation of beta3-adrenoceptors by selective agonists improves insulin action and stimulates energy metabolism in various rodent models of obesity and type 2 diabetes. Whether selective beta3-adrenoceptor stimulation exerts metabolic actions in humans remains to be proven. The effects of a highly selective beta3 adrenoceptor agonist on insulin action, energy metabolism, and body composition were assessed in 14 healthy young lean male volunteers (age 22.5 +/- 3.3 years, 15 +/- 5% body fat [mean +/- SD]) randomly assigned to 8 weeks of treatment with either 1,500 mg/day of CL 316,243 (n = 10) or placebo (n = 4). Insulin-mediated glucose disposal (IMGD), nonoxidative glucose disposal (NOGD), oxidative glucose disposal (OGD) (indirect calorimetry), and splanchnic glucose output (SGO; beta3 [H3]glucose) were determined during a 100-min hyperinsulinemic-euglycemic glucose clamp (40 mU x m(-2) x min(-1)) before and after 4 and 8 weeks of treatment. The 24-h energy expenditure (24-EE), 24-h respiratory quotient (24-RQ), and the oxidation rates of fat and carbohydrate were determined in a respiratory chamber before and after 8 weeks. After 4 weeks, treatment with CL 316,243 increased IMGD (+45%, P < 0.01) in a plasma concentration-dependent manner (r = 0.76, P < 0.02). This effect was due to an 82% increase in NOGD (P < 0.01), while OGD and SGO remained unchanged. The effects on insulin action were markedly diminished after 8 weeks; this was significantly related to an unexpected decline in the plasma concentrations of CL 316,243 (-36%, P = 0.08). At this time, 24-RQ was lowered (P < 0.001), corresponding to a 23% increase in fat oxidation (P < 0.01) and a 17% decrease in carbohydrate oxidation (P = 0.05). The 24-EE after 8 weeks did not differ from baseline, and there was no change in body weight or body composition. Plasma concentrations of glucose, insulin, and leptin were unaffected by treatment, while free fatty acid concentrations increased by 41% (P < 0.05), again linearly with the achieved plasma concentration of CL 316,243 (r = 0.67, P < 0.05). Treatment with CL 316,243 had no effect on heart rate or blood pressure and caused no cases of tremors. We conclude that treatment of lean male subjects with CL 316,243 increases insulin action and fat oxidation, both in a plasma concentration-dependent manner. This is the first study to demonstrate unequivocal metabolic effects of a highly selective beta3-adrenoceptor agonist in humans. PMID- 9753293 TI - Prolonged oxidative stress impairs insulin-induced GLUT4 translocation in 3T3-L1 adipocytes. AB - Prolonged exposure of 3T3-L1 adipocytes to micromolar concentrations of H2O2 results in an impaired response to the acute metabolic effects of insulin. In this study, we further characterized the mechanisms by which oxidative stress impairs insulin stimulation of glucose transport activity. Although insulin induced a 2.5-fold increase in plasma membrane GLUT4 content and a 50% reduction in its abundance in the low-density microsomal (LDM) fraction in control cells, oxidation completely prevented these responses. The net effect of insulin on 2 deoxyglucose uptake activity was reduced in oxidized cells and could be attributed to GLUT1 translocation. Insulin stimulation of insulin receptor substrate (IRS) 1 tyrosine phosphorylation and the association of IRS-1 with phosphatidylinositol (PI) 3-kinase were not impaired by oxidative stress. However, a 1.9-fold increase in the LDM content of the p85 subunit of PI 3-kinase after insulin stimulation was observed in control, but not in oxidized, cells. Moreover, although insulin induced an increase in IRS-1-associated PI 3-kinase activity in the LDM in control cells, this effect was prevented by oxidation. These findings suggest that prolonged low-grade oxidative stress impairs insulin stimulated GLUT4 translocation, potentially by interfering with compartment specific activation of PI 3-kinase. PMID- 9753294 TI - Prevention of diabetes in NOD mice by a mutated I-Ab transgene. AB - Susceptibility to the human autoimmune disease IDDM is strongly associated with those haplotypes of the major histocompatibility complex (MHC) carrying DQB1 alleles that do not encode aspartic acid at codon 57. Similarly, in a spontaneous animal model of this disease, the NOD mouse, the genes of the MHC play an important role in the development of diabetes. The DQB1 homolog in NOD mice, I Ab(g7), encodes a histidine at codon 56 and a serine at codon 57, while all other known I-Ab alleles encode proline and aspartic acid, respectively, at these positions. We therefore mutated the NOD I-Ab allele to encode proline at position 56 and aspartic acid at position 57 and introduced this allele onto the NOD genetic background to study the effect of these substitutions on susceptibility to diabetes. No transgenic mice developed diabetes by 8 months of age, and transgenic mice had markedly reduced lymphocytic infiltration in the pancreas compared with nontransgenic littermates. Furthermore, splenocytes from transgenic mice failed to proliferate or secrete gamma-interferon in response to a panel of beta-cell autoantigens, although the mice did produce beta-cell specific antibodies. Interestingly, the proportion of IgG1 and IgE relative to IgG2a comprising these autoantibodies was much greater in transgenic mice compared with nontransgenic control mice. Finally, T-cells from transgenic mice inhibited the adoptive transfer of diabetes to irradiated recipients. This inhibition was partially reversed by treatment of the recipients with a combination of anti interleukin (IL)-4 and anti-IL-10 monoclonal antibodies. Thus, a transgenic class II MHC allele encoding aspartic acid at B57 prevents diabetes, in part, by promoting the production of IL-4 and IL-10, which interfere with the effector phase of the diabetic process. PMID- 9753295 TI - Complementary action of antioxidant enzymes in the protection of bioengineered insulin-producing RINm5F cells against the toxicity of reactive oxygen species. AB - To determine the importance of different antioxidative enzymes for the defense status of insulin-producing cells, the effects of stable overexpression of glutathione peroxidase (Gpx), catalase (Cat), or Cu/Zn superoxide dismutase (SOD) in insulin-producing RINm5F cells on the cytotoxicity of hydrogen peroxide (H2O2), hypoxanthine/xanthine oxidase (H/XO), and menadione have been investigated. Single overexpression of Cat or Gpx provided less protection than the combined expression of Cat plus SOD or Cat plus Gpx, while single overexpression of SOD either had no effect on the toxicity of the test compounds or increased it. RINm5F cells were also susceptible to butylalloxan, a lipophilic alloxan derivative that is selectively toxic to pancreatic beta-cells. Overexpression of enzymes, both alone and in combination, did not protect against butylalloxan-induced toxicity while SOD overexpression increased it, as evident from a half maximally effective concentration (EC50) value. The addition of Cat to the culture medium completely prevented the toxic effects of H2O2 and H/XO but had no significant effect on the toxicity of menadione or butylalloxan. Extracellular SOD had no effect on the toxicity of any of the test compounds. The results of this study show the importance of a combination of antioxidant enzymes in protecting against the toxicity of reactive oxygen species. Thus, overexpression of Cat and Gpx, alone or in combination with SOD, by use of molecular biology techniques can protect insulin-producing cells against oxidative damage. This may represent a strategy to protect pancreatic beta-cells against destruction during the development of autoimmune diabetes and emphasizes the importance of optimal antioxidative enzyme equipment for protection against free radical-mediated diseases. PMID- 9753296 TI - Capillary blood pressure in syngeneic rat islets transplanted under the renal capsule is similar to that of the implantation organ. AB - The aim of the present study was to measure capillary blood pressure and interstitial pressure in transplanted pancreatic islets and to correlate these measurements to capillary and tubular pressures in the adjacent kidney. For this purpose, 250 syngeneic islets were implanted under the renal capsule of WF rats and studied 1, 2, or 6 months after transplantation. Some of the animals studied after 1 and 2 months were streptozotocin (STZ)-induced diabetic. Measurements were performed during basal conditions or after an acute glucose-stimulation of insulin release. The hydrostatic pressures were determined in vivo by direct micropuncture. The islet transplant capillary pressure in normoglycemic animals was 6.9 +/- 0.4 mmHg (n = 9), 10.0 +/- 0.8 mmHg (n = 7), and 12.4 +/- 0.8 mmHg (n = 7) when measured 1, 2, and 6 months after implantation, respectively. Previous data from our laboratory showed that the normal capillary pressure of native rat pancreatic islets is approximately 3 mmHg. The blood pressure in kidney peritubular capillaries was 10-12 mmHg in both transplanted and control animals. Islet transplant interstitial pressures were 4-6 mmHg in the normoglycemic recipients at 1, 2, and 6 months after transplantation. Acute glucose stimulation had no effect on islet transplant interstitial pressure or peritubular or transplant capillary blood pressures. Capillary pressures in the islet grafts were slightly increased 1 month after transplantation in STZ-induced diabetic rats, and this was associated with an increased blood perfusion of the transplants. However, 2 months after transplantation there were no differences in transplant capillary blood pressure between diabetic and normoglycemic animals. The graft interstitial pressure was, on the contrary, decreased in the diabetic animals 2 months after transplantation. We concluded that the capillary blood pressure in islets implanted under the renal capsule was similar to that of the implantation organ, which was three to four times higher than that normally found in native islets. PMID- 9753297 TI - HLA-DR binding analysis of peptides from islet antigens in IDDM. AB - HLA molecules are essential for thymic education and HLA restriction of T-cell responses. We therefore analyzed the HLA-DR binding affinities of synthetic peptides covering the entire sequences of GAD65, islet cell antigen 69 (ICA69), and (pro)insulin, which are candidate antigens in the autoimmune process of T cell-mediated destruction of the pancreatic beta-cells. Subsequently, peptide HLA DR binding was correlated to peptide antigenicity by comparing known T-cell epitopes with their HLA-binding affinities defined in this study. The results demonstrate the following. 1) (Pro)insulin peptides display a strong binding affinity for HLA-DR2, which is associated with negative genetic predisposition to IDDM, whereas poor binding was observed for HLA-DR molecules neutrally or positively associated with IDDM. This suggests that the absence of insulin reactive T-cells in DR2+ individuals may be explained by negative selection on high-affinity DR2 binding insulin peptides. 2) Most autoantigenic peptides display promiscuous HLA-DR binding patterns. This promiscuity in itself is not sufficient to explain the genetic association of HLA-DR with development of IDDM. 3) HLA-DR3 binding of autoantigenic GAD65 peptides is relatively weak compared with that of other known T-cell epitopes. 4) All peptide epitopes recognized by HLA-DR-restricted T-cells from either IDDM patients or GAD65-immunized HLA-DR transgenic mice bind with high affinity to their HLA-DR restriction molecule (P < 0.0006). In contrast, T-cell epitopes recognized by nondiabetic controls bind DR molecules with weak or undetectable affinity. These results thus indicate a strong correlation between antigenicity and HLA-DR binding affinity of GAD65 peptides in IDDM. Furthermore, negative thymic selection of insulin peptides in low-risk (HLA-DR2 expressing) subjects may explain the lack of autoreactivity to insulin in such individuals. PMID- 9753298 TI - Ciliary neurotrophic factor potentiates the beta-cell inhibitory effect of IL 1beta in rat pancreatic islets associated with increased nitric oxide synthesis and increased expression of inducible nitric oxide synthase. AB - Proinflammatory cytokines are implicated as effector molecules in the pathogenesis of IDDM. Interleukin-6 (IL-6) alone or in combination with IL-1beta inhibits glucose-stimulated insulin release from isolated rat pancreatic islets by unknown mechanisms. Here we investigated 1) if the effects of IL-6 are mimicked by ciliary neurotrophic factor (CNTF), another member of the IL-6 family of cytokines signaling via gp130, 2) the possible cellular mechanisms for these effects, and 3) if islet endocrine cells are a source of CNTF. CNTF (20 ng/ml) potentiated IL-1beta-mediated (5-150 pg/ml) nitric oxide (NO) synthesis from neonatal Wistar rat islets by 31-116%, inhibition of accumulated insulin release by 34-49%, and inhibition insulin response to a 2-h glucose challenge by 31-36%. CNTF potentiated IL-1beta-mediated NO synthesis from RIN-5AH cells by 83%, and IL 1beta induced islet inducible NO-synthase (iNOS) mRNA expression fourfold. IL-6 (10 ng/ml) also potentiated IL-1beta-mediated NO synthesis and inhibition of insulin release, whereas beta-nerve growth factor (NGF) (5 or 50 ng/ml) had no effect. mRNA for CNTF was expressed in rat islets and in islet cell lines. In conclusion, CNTF is constitutively expressed in pancreatic beta-cells and potentiates the beta-cell inhibitory effect of IL-1beta in association with increased iNOS expression and NO synthesis, an effect shared by IL-6 but not by beta-NGF. These findings indicate that signaling via gp130 influences islet NO synthesis associated with iNOS expression. We hypothesize that CNTF released from destroyed beta-cells during the inflammatory islet lesion leading to IDDM may potentiate IL-1beta action on the beta-cells. PMID- 9753299 TI - Acute lowering of plasma fatty acids lowers basal insulin secretion in diabetic and nondiabetic subjects. AB - The objective of this study was to determine whether basal plasma free fatty acid (FFA) concentrations affect basal insulin secretion rates (ISRs). Effects of FFA levels on basal ISRs were evaluated by lowering basal plasma FFA levels with nicotinic acid (NA) (100-150 mg p.o., q 30 min x 4 h) in type 2 diabetic patients and in normal volunteers. Lowering of FFAs (from approximately 600 to approximately 100 micromol/l) lowered ISRs in type 2 diabetic patients during isoglycemic clamping (from 139 to 101 pmol/min; -23%; P < 0.02) and euglycemic clamping (from 99 to 63 pmol/min; -36%; P < 0.03) and in normal subjects during euglycemic clamping (from 127 to 96 pmol/min; -25%; P < 0.03). In addition, peripheral insulin concentrations decreased by approximately 30% in diabetic and nondiabetic subjects. NA had no direct effect on ISRs; that is, NA did not change ISRs when plasma FFAs were prevented from decreasing with a lipid/heparin infusion. We concluded that 1) basal plasma FFAs exerted physiologically important, long-lasting effects supporting 25-33% of basal insulin secretion in nondiabetic and diabetic subjects; 2) basal plasma FFAs were responsible for some of the hyperinsulinemia in normoglycemic obese subjects; and 3) NA had no direct effect on insulin secretion. PMID- 9753301 TI - Glycated cholecystokinin-8 has an enhanced satiating activity and is protected against enzymatic degradation. AB - Monoglycated cholecystokinin octapeptide (CCK-8) (glucitol-Asp1 adduct) modified at the NH2-terminus was prepared under hyperglycemic conditions, purified by high performance liquid chromatography, and characterized by mass spectrometry (Mr 1228.4 Da) and peptide sequencing. CCK-8 (100 nmol/kg, i.p.) significantly (P < 0.001) reduced voluntary food intake of fasted mice for up to 30 min after its administration, compared with saline-administered controls. Glycated CCK-8 reduced food intake at 30-120 min (P < 0.01 to P < 0.001) and significantly reduced feeding compared with CCK-8 from 60 to 120 min (P < 0.01). In vitro plasma degradation studies indicated that glycated CCK-8 was resistant to the normal rapid enzymatic conversion to CCK fragments. This study demonstrated that CCK-8 is a potent short-term inhibitor of food intake, and that structural modification of this peptide by amino-terminal glycation leads to enhanced satiating activity, partially due to increased resistance to serum aminopeptidase degradation. PMID- 9753300 TI - Circulating fatty acids are essential for efficient glucose-stimulated insulin secretion after prolonged fasting in humans. AB - In the fasted rat, efficient glucose-stimulated insulin secretion (GSIS) is absolutely dependent on an elevated level of circulating free fatty acids (FFAs). To determine if this is also true in humans, nonobese volunteers were fasted for 24 h (n = 5) or 48 h (n = 5), after which they received an infusion of either saline or nicotinic acid (NA) to deplete their plasma FFA pool, followed by an intravenous bolus of glucose. NA treatment resulted in a fall in basal insulin concentrations of 35 and 45% and in the area under the insulin response curve (area under the curve [AUC]) to glucose of 47 and 42% in the 24- and 48-h fasted individuals, respectively. The 48-h fasted subjects underwent the same procedure with the addition of a coinfusion of Intralipid plus heparin (together with NA) to maintain a high concentration of plasma FFAs throughout the study. The basal level and AUC for insulin were now completely normalized (C-peptide profiles paralleled those for insulin). To assess the effect of an overnight fast, nonobese (n = 6) and obese (n = 6) subjects received an infusion of either saline or NA, followed by a hyperglycemic clamp (200 mg/dl). The insulin AUC in response to glucose was unaffected by lowering of the FFA level in nonobese subjects, but fell by 29% in the obese group. The data clearly demonstrate that in humans, the rise in circulating FFA levels after 24 and 48 h of food deprivation is critically important for pancreatic beta-cell function both basally and during subsequent glucose loading. They also suggest that the enhancement of GSIS by FFAs in obese individuals is more prominent than that seen in their nonobese counterparts. PMID- 9753302 TI - Increased OB gene expression leads to elevated plasma leptin concentrations in patients with chronic primary hyperinsulinemia. AB - Leptin, a hormone secreted by adipocytes, decreases food intake and increases energy expenditure. The role of insulin in the regulation of leptin secretion is poorly understood and is still a topic of debate. Insulin increases leptin mRNA synthesis in rodents, but in humans, the available data are discordant. To investigate the role of chronic hyperinsulinemia in the regulation of plasma leptin concentrations, we studied 13 patients with surgically confirmed insulinoma before and after tumor removal, along with 15 healthy control subjects matched for sex, age, and BMI. Immunoreactive plasma leptin levels were measured by radioimmunoassay; leptin mRNA levels were also determined by reverse transcription-competitive polymerase chain reaction in a subgroup of six patients with insulinoma and six control subjects. All determinations were made with subjects in the fasting state. Plasma leptin concentrations correlated positively with leptin mRNA levels (r = 0.880, P < 0.001). Leptin levels, both plasma protein and mRNA, were significantly higher in the insulinoma patients than in the control subjects (plasma protein: 17.5 +/- 3.6 vs. 2.9 +/- 0.4 ng/ml, respectively, P < 0.001; mRNA: 0.98 +/- 0.33 vs. 0.19 +/- 0.064 amol/microg RNA, respectively, P < 0.05), and they correlated positively with fasting plasma insulin levels in the patients with insulinoma (plasma protein: r = 0.686, P < 0.01; mRNA: 0.796, P < 0.05). Finally, removal of the insulin-secreting tumor was followed by the normalization of plasma leptin levels. In summary, in patients with insulinoma, 1) plasma leptin levels and leptin mRNA are elevated; 2) a direct relationship exists between leptin, both circulating protein and mRNA, and insulin concentrations; and 3) plasma leptin returns to normal levels after tumor removal. These data, therefore, support a role for insulin in the chronic regulation of leptin gene expression. PMID- 9753303 TI - Plasma glucose levels are reduced in rats and mice treated with an inhibitor of glucose-6-phosphate translocase. AB - The activity of glucose-6-phosphatase (G-6-Pase) in isolated rat microsomes was inhibited by a new selective inhibitor of the multi-subunit G-6-Pase system, 1-[2 (4-chloro-phenyl)-cyclopropylmethoxy]-3,4-dihydroxy-5-(3-imid azo[4,5-b]pyridin-1 yl-3-phenyl-acryloyloxy)-cyclohexanecarboxylic acid (compound A) with a 50% inhibitory concentration (IC50) of approximately 10 nmol/l. Compound A (500 nmol/l) inhibited the uptake of [14C]glucose-6-phosphate (G-6-P) into intact isolated rat microsomes, confirming that this agent blocks G-6-P translocation, as suggested by previous studies using intact and permeabilized microsomes. The inhibition of microsomal G-6-P transport by compound A was associated with inhibition of the rate of glucose output from rat hepatocytes incubated in the presence of 25 nmol/l glucagon (IC50 approximately 320 nmol/l.) Compound A (1 micromol/l) also inhibited the basal rate of glucose production by rat hepatocytes by 47%. Intraperitoneal administration of compound A to fasted mice lowered circulating plasma glucose concentrations dose-dependently at doses as low as 1 mg/kg. This effect was comparatively short-lived; glucose lowering was maximal at 30 min after dosing with 100 mg/kg compound A (-71%) and declined thereafter, being reversed within 3 h. A similar time course of glycemic response was observed in fasted rats; glucose lowering was maximal 30 min after dosing with 100 mg/kg compound A (-36%) and declined until the effect was fully reversed by 3 h postdose. In rats subjected to compound A treatment, liver glycogen content was increased. G-6-P and lactate levels were maximally elevated 30 min after dosing and declined thereafter. Cumulatively, these results suggest that the mechanism of glucose lowering by compound A was via inhibition of G-6-Pase activity, mediated through inhibition of the T1 subunit of the microsomal G-6 Pase enzyme system. Drug levels measured over the same time course as that used to assess in vivo efficacy peaked within 30 min of administration, then declined, which is consistent with the transient changes in plasma glucose and liver metabolites. PMID- 9753305 TI - Prevalence of insulin resistance in metabolic disorders: the Bruneck Study. AB - The prevalence of insulin resistance in the most common metabolic disorders is still an undefined issue. We assessed the prevalence rates of insulin resistance in subjects with impaired glucose tolerance (IGT), NIDDM, dyslipidemia, hyperuricemia, and hypertension as identified within the frame of the Bruneck Study. The study comprised an age- and sex-stratified random sample of the general population (n = 888; aged 40-79 years). Insulin resistance was estimated by homeostasis model assessment (HOMA(IR)), preliminarily validated against a euglycemic-hyperinsulinemic clamp in 85 subjects. The lower limit of the top quintile of HOMA(IR) distribution (i.e., 2.77) in nonobese subjects with no metabolic disorders (n = 225) was chosen as the threshold for insulin resistance. The prevalence of insulin resistance was 65.9% in IGT subjects, 83.9% in NIDDM subjects, 53.5% in hypercholesterolemia subjects, 84.2% in hypertriglyceridemia subjects, 88.1% in subjects with low HDL cholesterol, 62.8% in hyperuricemia subjects, and 58.0% in hypertension subjects. The prevalence of insulin resistance in subjects with the combination of glucose intolerance (IGT or NIDDM), dyslipidemia (hypercholesterolemia and/or hypertriglyceridemia and/or low HDL cholesterol), hyperuricemia, and hypertension (n = 21) was 95.2%. In isolated hypercholesterolemia, hypertension, or hyperuricemia, prevalence rates of insulin resistance were not higher than that in nonobese normal subjects. An appreciable number of subjects (n = 85, 9.6% of the whole population) was insulin resistant but free of IGT, NIDDM, dyslipidemia, hyperuricemia, and hypertension. These results from a population-based study documented that 1) in hypertriglyceridemia and a low HDL cholesterol state, insulin resistance is as common as in NIDDM, whereas it is less frequent in hypercholesterolemia, hyperuricemia, and hypertension; 2) the vast majority of subjects with multiple metabolic disorders are insulin resistant; 3) in isolated hypercholesterolemia, hyperuricemia, or hypertension, insulin resistance is not more frequent than can be expected by chance alone; and 4) in the general population, insulin resistance can be found even in the absence of any major metabolic disorders. PMID- 9753304 TI - Peripheral neuropathy in transgenic diabetic mice: restoration of C-fiber function with human recombinant nerve growth factor. AB - Mice (Ins.Dd1) with hypoinsulinemic diabetes were created by increased expression of syngeneic major histocompatibility complex (MHC) class I protein in pancreatic beta-cells. The diabetic state was characterized in these mice by high glucose concentrations and islet pathology. To determine whether a neuropathy would develop, motor and sensory conduction velocities (CV) were determined in the sciatic nerves of 2-, 4-, and 7-month-old control and diabetic littermate male mice. Recording bipolar electrodes were placed in the plantar muscles of the hind foot of anesthetized (ketamine/xylazine) mice. Bipolar stimulating electrodes were positioned near the sciatic nerve at the sciatic notch or near the tibial nerve at the ankle. Motor CV from alpha-motor fibers and sensory CV from proprioceptive Aalpha nerves were measured and expressed as meters per second (m/s). Group data are reported as mean +/- SE and compared by analysis of variance. The CVs from nondiabetic mice (controls) were not different across the three ages and averaged 41.3 +/- 1.7 m/s for motor and 38.7 +/- 1.7 m/s for sensory. The motor CVs from diabetic mice at 2 and 4 months were similar to controls. Sensory CVs were unchanged at 2 months but were lower at 4 months (18.9 +/- 2.4 m/s). Both sensory (23.9 +/- 2.1 m/s) and motor (18.9 +/- 1.8 m/s) CVs were significantly reduced at 7 months, which is indicative of a polyneuropathy. NGF has well-known trophic effects on sympathetic and small sensory neurons. To determine whether NGF could influence this neuropathy, 6-month-old control and diabetic mice were divided into the following groups: 1) control + vehicle, 2) diabetic + vehicle, and 3) diabetic + NGF (1 mg/kg, 3x week, s.c.). After 1 month of treatment, motor and sensory CVs were determined. In some mice, the branches of the sciatic nerve were exposed and in situ recordings from the sural nerve were performed to determine compound C-fiber CV, integral, and amplitude. Sensory CV, determined via Hoffmann's reflex (H-reflex) (A-fiber), was decreased in diabetic compared with control animals as expected (P < 0.05), and NGF did not alter this parameter. Continuing diabetes reduced the amplitude (0.9 +/- 0.2 vs. 3.2 +/- 0.7 mV x 10(-2); P < 0.05) and integral (6.9 +/- 1.9 mV/ms vs. 18.8 +/- 4.4 mV/ms; P < 0.05) of the C-fiber response versus control, suggesting fiber loss. NGF treatment normalized C-fiber amplitude (2.9 +/- 0.8 mV x 10(-2)) and integral (21.2 +/- 6.5 mV/ms) in animals with established diabetes, with no effect on blood glucose. The C-fiber CV was similar in all groups, indicating that the animals had some normally conducting small fiber sensory nerves. These studies characterized a motor and sensory polyneuropathy in transgenic diabetic mice and are the first to demonstrate directly that NGF treatment can protect or restore abnormal sensory C-fiber function. PMID- 9753306 TI - Isolation and characterization of the human PAX4 gene. PMID- 9753307 TI - No association between the Friedreich's ataxia gene and NIDDM in the French population. PMID- 9753308 TI - Gln27Glu variant of the human beta2-adrenoreceptor gene is not associated with early-onset obesity in Danish men. PMID- 9753309 TI - Oral contraceptives and thrombosis: an overview of study methods and recent results. AB - Studies of the associations between oral contraceptives and cardiovascular disease are limited by the extreme rarity of these problems among young women. There are no randomized, controlled trials, and the large, prospective cohort studies only have data regarding older oral contraceptive formulations that are now little used. Historical cohort studies that use record-linkage techniques to analyze data from computerized databases represent a new approach to assessing oral contraceptive use and thrombosis. The largest new studies, with the most sophisticated analyses, use the case-control design. A surprising result of the new studies was a difference in risk of thrombosis according to progestin type. Because these are observational rather than randomized studies, clinical factors influence the choice of oral contraceptive and may bias the study results. Controversy about the surprising results has stimulated additional analyses and critical reviews in an attempt to explain the associations. PMID- 9753310 TI - Bias versus causality: interpreting recent evidence of oral contraceptive studies. AB - Late in 1995 and early 1996, 4 epidemiologic studies were published that resulted in a crude mean weighted relative risk of approximately 2 when third-generation oral contraceptives were compared with second-generation oral contraceptives as risk factors for venous thromboembolism. This article reviews empirical evidence on bias or systematic error that may have influenced the estimates of association. The Bradford-Hill criteria to distinguish causality from an observed association were used to consider whether third-generation oral contraceptives cause an apparent excess in the occurrence of venous thromboembolism. Bias is more likely than a causal relationship to explain the associations observed for venous thromboembolism. For myocardial infarction, bias may mask the full benefit of third-generation oral contraceptives. For stroke, the question of causality is moot because statistically significant differences between third- and second generation products have not been detected. The clinical importance and the public health significance of any differences among the various products with respect to adverse cardiovascular outcomes are trivial and undetectable because of the extremely low incidence of those disorders among users of oral contraceptives. The oral contraceptive pill is 99.9% effective when used correctly. All oral contraceptives on the market are safe and getting safer. PMID- 9753311 TI - Effects on hemostatic variables of desogestrel- and gestodene-containing oral contraceptives in comparison with levonorgestrel-containing oral contraceptives: a review. AB - In some studies third-generation oral contraceptives have been reported to be associated with a higher risk of venous thromboembolism than are second generation oral contraceptives, whereas recent, more refined studies have not confirmed this. The reasons for the alleged differences are under discussion, and differential effects on hemostasis have been proposed. Eighteen studies comparing second- and third-generation oral contraceptives with respect to their effects on hemostasis were analyzed. Significant changes from baseline were reported for many variables with both second- and third-generation oral contraceptives without significant between-group differences. Also, in a combined analysis of nonsignificant changes, no consistent pattern of change emerged for any marker, with the exception of higher factor VII levels associated with third-generation oral contraceptives. However, factor VII is not related to venous thromboembolism risk. In addition, 1 cross-sectional study with an unvalidated assay reported a higher ratio of activated protein C sensitivity with third-generation oral contraceptives. Only 2 components of the hemostatic system (factor VII and activated protein C sensitivity ratio) emerged as potentially differentially affected by second- and third-generation oral contraceptives; the association with venous thromboembolism risk is questionable in the former case and unknown in the latter. PMID- 9753312 TI - Thrombotic diseases in young women and the influence of oral contraceptives. AB - OBJECTIVE: In the evaluation of the clinical impact of thrombotic diseases in young women, age-specific incidence rates must be calculated for both arterial and venous thrombotic diseases, but also the case-fatality rate and figures for the clinical consequences among those who survive thrombosis must be included. The aim of this analysis was to quantify the clinical impacts of both arterial and venous thrombotic diseases among young, nonpregnant women and thereafter to assess the influences of oral contraceptives on these measures. STUDY DESIGN: Nationwide register data on the morbidity and mortality of venous thromboembolism, myocardial infarction, and thrombotic stroke in Denmark, 1980 1993, and 3 ongoing case-control studies to assess the influence of oral contraceptives on the risk for development of these thrombotic diseases. RESULTS: In women 15-29 years old venous thromboembolism is about twice as common as arterial complications, whereas in women 30-44 years old the number of arterial complications exceeds that of venous diseases by about 50%. The mortality rate from arterial diseases is 3.5 times higher than that from venous diseases among women <30 years old and 8.5 times higher than that from venous diseases in women 30-44 years old. The proportion of women with a significant disability among women who had an arterial complication was about 30%; the proportion was about 5% among women with venous thromboembolism. CONCLUSION: Anticipating a differential influence on venous and arterial diseases from oral contraceptives with second- and third-generation progestogens, it was calculated that users of oral contraceptives with second-generation progestogens had 30% greater increased risk of thrombotic diseases, 260% greater increased risk of thrombotic deaths, and 220% greater increased risk of thrombotic disability than users of oral contraceptives with third-generation progestogens. PMID- 9753313 TI - Myocardial infarction and stroke in young women: what is the impact of oral contraceptives? AB - Recent discussions have centered on the small apparent risk increase for venous thromboembolism found with newer oral contraceptives (third-generation oral contraceptives containing the progestins desogestrel and gestodene) compared with older oral contraceptives (second-generation). This article reviews the studies addressing the association between oral contraceptive use and thromboembolic conditions affecting the arterial system, ischemic stroke, and myocardial infarction. Differences are found between a US database study, which showed no risk of ischemic stroke or myocardial infarction associated with low-dose oral contraceptive use, and the European studies, which showed oral contraceptive use in general to be associated with increased risks of ischemic stroke and myocardial infarction. The European studies showed no difference between oral contraceptive generations with respect to the occurrence of ischemic stroke; however, the risk of myocardial infarction associated with oral contraceptive use was consistently lower for third- than for second-generation oral contraceptives. Although there seems to be no differential risk of ischemic stroke associated with oral contraceptive generations, third-generation oral contraceptives appear to be consistently associated with no excess risk of myocardial infarction. In all instances, however, cardiovascular risk factors other than oral contraceptive use play the predominant role in the occurrence of ischemic stroke and myocardial infarction. PMID- 9753314 TI - Oral contraceptives and venous thromboembolic disease: the findings from database studies in the United Kingdom and Germany. AB - OBJECTIVE: Three research articles published in late 1995 and early 1996 suggested that oral contraceptives containing either of the newer progestogens (gestodene or desogestrel) could be associated with an increased risk of venous thromboembolism. During the months after the initial publications, the results have been scrutinized with great care and further studies have been published. The findings of 2 recent database studies, 1 in the United Kingdom and 1 in Germany, are presented in this article. PATTERNS OF USE: The average age of users of combined oral contraceptives in Germany was 27 years, compared with 26 years in the United Kingdom. In Germany the use of gestodene-based products was lower than that in the United Kingdom. In the United Kingdom the users of desogestrel with 20 microg ethinyl estradiol (Mercilon) were older than the users of desogestrel with 30 microg ethinyl estradiol (Marvelon). CRUDE INCIDENCE: The crude incidence of venous thromboembolism in the UK study was 4.1 cases/10,000 woman-y exposure to combined oral contraceptives. In Germany it was 4.2 cases/10,000 woman-y. In Germany the rates among users of second-generation combined oral contraceptives were higher than those among users of third generation products. The reverse was the case in the United Kingdom. In the United Kingdom the crude incidence rates were higher for the 20 microg estrogen desogestrel product than for the 30 microg product. CASE-CONTROL ANALYSIS: The adjusted odds ratios in the UK study did not show significant increases for desogestrel or gestodene compared with levonorgestrel products. There were inconsistencies in the results among centers in the 2 international studies (the World Health Organization and Transnational studies). In both there was a consistent inverse dose-response relationship with estrogen in all centers. CONCLUSION: The limitations of the observational studies are such that the hypothesis that the newer progestogens are more likely to cause venous thromboembolism cannot be proved. PMID- 9753315 TI - Sorting out genes that regulate epithelial and neuronal polarity. PMID- 9753316 TI - Apoptotic pathways: the roads to ruin. PMID- 9753317 TI - RuvA gets X-rayed on holliday. PMID- 9753318 TI - Smad3 mutant mice develop metastatic colorectal cancer. AB - TGFbeta-related growth factors have been implicated in a variety of developmental and physiological processes in organisms ranging from nematodes to mammals. TGFbeta transduces its signal to the interior of the cell via Smad2, Smad3, and Smad4. We report the cloning and targeted disruption of the mouse Smad3 gene. Smad3 mutant mice are viable and fertile. Between 4 and 6 months of age, the Smad3 mutant mice become moribund with colorectal adenocarcinomas. The neoplasms penetrate through the intestinal wall and metastasize to lymph nodes. These results directly implicate TGFbeta signaling in the pathogenesis of colorectal cancer and provide a compelling animal model for the study of human colorectal cancer. PMID- 9753319 TI - Tumor induction of VEGF promoter activity in stromal cells. AB - We have established a line of transgenic mice expressing the A. victoria green fluorescent protein (GFP) under the control of the promoter for vascular endothelial growth factor (VEGF). Mice bearing the transgene show green cellular fluorescence around the healing margins and throughout the granulation tissue of superficial ulcerative wounds. Implantation of solid tumors in the transgenic mice leads to an accumulation of green fluorescence resulting from tumor induction of host VEGF promoter activity. With time, the fluorescent cells invade the tumor and can be seen throughout the tumor mass. Spontaneous mammary tumors induced by oncogene expression in the VEGF-GFP mouse show strong stromal, but not tumor, expression of GFP. In both wound and tumor models the predominant GFP positive cells are fibroblasts. The finding that the VEGF promoter of nontransformed cells is strongly activated by the tumor microenvironment points to a need to analyze and understand stromal cell collaboration in tumor angiogenesis. PMID- 9753320 TI - Apaf1 (CED-4 homolog) regulates programmed cell death in mammalian development. AB - The cytosolic protein APAF1, human homolog of C. elegans CED-4, participates in the CASPASE 9 (CASP9)-dependent activation of CASP3 in the general apoptotic pathway. We have generated by gene trap a null allele of the murine Apaf1. Homozygous mutants die at embryonic day 16.5. Their phenotype includes severe craniofacial malformations, brain overgrowth, persistence of the interdigital webs, and dramatic alterations of the lens and retina. Homozygous embryonic fibroblasts exhibit reduced response to various apoptotic stimuli. In situ immunodetection shows that the absence of Apaf1 protein prevents the activation of Casp3 in vivo. In agreement with the reported function of CED-4 in C. elegans, this phenotype can be correlated with a defect of apoptosis. Our findings suggest that Apaf1 is essential for Casp3 activation in embryonic brain and is a key regulator of developmental programmed cell death in mammals. PMID- 9753321 TI - Apaf1 is required for mitochondrial pathways of apoptosis and brain development. AB - Apoptosis is essential for the precise regulation of cellular homeostasis and development. The role in vivo of Apaf1, a mammalian homolog of C. elegans CED-4, was investigated in gene-targeted Apaf1-/- mice. Apaf1-deficient mice exhibited reduced apoptosis in the brain and striking craniofacial abnormalities with hyperproliferation of neuronal cells. Apaf1-deficient cells were resistant to a variety of apoptotic stimuli, and the processing of Caspases 2, 3, and 8 was impaired. However, both Apaf1-/- thymocytes and activated T lymphocytes were sensitive to Fas-induced killing, showing that Fas-mediated apoptosis in these cells is independent of Apaf1. These data indicate that Apaf1 plays a central role in the common events of mitochondria-dependent apoptosis in most death pathways and that this role is critical for normal development. PMID- 9753322 TI - LIN-10 is a shared component of the polarized protein localization pathways in neurons and epithelia. AB - We tested the model that neurons and epithelial cells use a shared mechanism for polarized protein sorting by comparing the pathways for localizing basolateral and postsynaptic proteins in C. elegans. GLR-1 glutamate receptors are localized to postsynaptic elements of central synapses and, when ectopically expressed, to basolateral membranes of epithelial cells. Proper localization of GLR-1 in both neurons and epithelia requires the PDZ protein LIN-10, defining LIN-10 as a shared component of the basolateral and postsynaptic localization pathways. Changing the GLR-1 carboxy-terminal sequence from a group I PDZ-binding consensus (-TAV) to a group II consensus (-FYV) restores GLR-1 synaptic localization in lin 10 mutants. Thus, these interneurons utilize at least two separate postsynaptic localization pathways. PMID- 9753323 TI - The LIN-2/LIN-7/LIN-10 complex mediates basolateral membrane localization of the C. elegans EGF receptor LET-23 in vulval epithelial cells. AB - In C. elegans, the LET-23 receptor tyrosine kinase is localized to the basolateral membranes of polarized vulval epithelial cells. lin-2, lin-7, and lin 10 are required for basolateral localization of LET-23, since LET-23 is mislocalized to the apical membrane in lin-2, lin-7, and lin-10 mutants. Yeast two-hybrid, in vitro binding, and in vivo coimmunoprecipitation experiments show that LIN-2, LIN-7, and LIN-10 form a protein complex. Furthermore, compensatory mutations in lin-7 and let-23 exhibit allele-specific suppression of apical mislocalization and signaling-defective phenotypes. These results present a mechanism for basolateral localization of LET-23 receptor tyrosine kinase by direct binding to the LIN-2/LIN-7/LIN-10 complex. Each of the binding interactions within this complex is conserved, suggesting that this complex may also mediate basolateral localization in mammals. PMID- 9753324 TI - A tripartite protein complex with the potential to couple synaptic vesicle exocytosis to cell adhesion in brain. AB - We identify a complex of three proteins in brain that has the potential to couple synaptic vesicle exocytosis to neuronal cell adhesion. The three proteins are: (1) CASK, a protein related to MAGUKs (membrane-associated guanylate kinases); (2) Mint1, a putative vesicular trafficking protein; and (3) Veli1, -2, and -3, vertebrate homologs of C. elegans LIN-7. CASK, Mint1, and Velis form a tight, salt-resistant complex that can be readily isolated. CASK, Mint1, and Velis contain PDZ domains in addition to other modules. However, no PDZ domains are involved in complex formation, leaving them free to recruit cell adhesion molecules, receptors, and channels to the complex. We propose that the tripartite complex acts as a nucleation site for the assembly of proteins involved in synaptic vesicle exocytosis and synaptic junctions. PMID- 9753325 TI - Cdc2 kinase directly phosphorylates the cis-Golgi matrix protein GM130 and is required for Golgi fragmentation in mitosis. AB - Mitotic fragmentation of the Golgi apparatus can be largely explained by disruption of the interaction between GM130 and the vesicle-docking protein p115. Here we identify a single serine (Ser-25) in GM130 as the key phosphorylated target and Cdc2 as the responsible kinase. MEK1, a component of the MAP kinase signaling pathway recently implicated in mitotic Golgi fragmentation, was not required for GM130 phosphorylation or mitotic fragmentation either in vitro or in vivo. We propose that Cdc2 is directly involved in mitotic Golgi fragmentation and that signaling via MEK1 is not required for this process. PMID- 9753326 TI - Signal sequence recognition in posttranslational protein transport across the yeast ER membrane. AB - We have analyzed how the signal sequence of prepro-alpha-factor is recognized during the first step of posttranslational protein transport into the yeast endoplasmic reticulum. Cross-linking studies indicate that the signal sequence interacts in a Kar2p- and ATP-independent reaction with Sec61p, the multispanning membrane component of the protein-conducting channel, by intercalation into transmembrane domains 2 and 7. While bound to Sec61p, the signal sequence forms a helix that is contacted on one side by Sec62p and Sec71p. The binding site is located at the interface of the protein channel and the lipid bilayer. Signal sequence recognition in cotranslational translocation in mammals appears to occur similarly. These results suggest a general mechanism by which the signal sequence could open the channel for polypeptide transport. PMID- 9753327 TI - Developmentally regulated Xist promoter switch mediates initiation of X inactivation. AB - Developmental regulation of the mouse Xist gene at the onset of X chromosome inactivation is mediated by RNA stabilization. Here, we show that alternate promoter usage gives rise to distinct stable and unstable RNA isoforms. Unstable Xist transcript initiates at a novel upstream promoter, whereas stable Xist RNA is transcribed from the previously identified promoter and from a novel downstream promoter. Analysis of cells undergoing X inactivation indicates that a developmentally regulated promoter switch mediates stabilization and accumulation of Xist RNA on the inactive X chromosome. PMID- 9753328 TI - The structure of supercoiled intermediates in DNA replication. AB - We studied the structure of replication intermediates accumulated by Tus-induced arrest of plasmid DNA replication at termination sites. For intermediates generated both in vitro with purified components and in vivo, superhelical stress is distributed throughout the entire partially replicated molecule; daughter DNA segments are wound around each other, and the unreplicated region is supercoiled. Thus, unlinking of parental DNA strands by topoisomerases can be carried out both behind and in front of the replication fork. We explain why previous studies with prokaryotic and eukaryotic replication intermediates discerned only supercoiling in the unreplicated portion. PMID- 9753329 TI - Structure of type IIbeta phosphatidylinositol phosphate kinase: a protein kinase fold flattened for interfacial phosphorylation. AB - Phosphoinositide kinases play central roles in signal transduction by phosphorylating the inositol ring at specific positions. The structure of one such enzyme, type IIbeta phosphatidylinositol phosphate kinase, reveals a protein kinase ATP-binding core and demonstrates that all phosphoinositide kinases belong to one superfamily. The enzyme is a disc-shaped homodimer with a 33 x 48 A basic flat face that suggests an electrostatic mechanism for plasma membrane targeting. Conserved basic residues form a putative phosphatidylinositol phosphate specificity site. The substrate-binding site is open on one side, consistent with dual specificity for phosphatidylinositol 3- and 5-phosphates. A modeled complex with membrane-bound substrate and ATP shows how a phosphoinositide kinase can phosphorylate its substrate in situ at the membrane interface. PMID- 9753330 TI - Three-dimensional structure of an evolutionarily conserved N-terminal domain of syntaxin 1A. AB - Syntaxin 1A plays a central role in neurotransmitter release through multiple protein-protein interactions. We have used NMR spectroscopy to identify an autonomously folded N-terminal domain in syntaxin 1A and to elucidate its three dimensional structure. This 120-residue N-terminal domain is conserved in plasma membrane syntaxins but not in other syntaxins, indicating a specific role in exocytosis. The domain contains three long alpha helices that form an up-and-down bundle with a left-handed twist. A striking residue conservation is observed throughout a long groove that is likely to provide a specific surface for protein protein interactions. A highly acidic region binds to the C2A domain of synaptotagmin I in a Ca2+-dependent interaction that may serve as an electrostatic switch in neurotransmitter release. PMID- 9753331 TI - Discordant organ xenotransplantation in primates: world experience and current status. AB - The pig-to-primate model is increasingly being utilized as the final preclinical means of assessing therapeutic strategies aimed at allowing discordant xenotransplantation. We review here the world experience of both pig-to-human and pig-to-nonhuman primate organ transplantation. Eight whole organ transplants using discordant mammalian donors have been carried out in human recipients; only one patient was reported (in 1923) to have survived for longer than 72 hr. Therapeutic approaches in the experimental laboratory setting have included pharmacologic immunosuppression, antibody and/or complement depletion or inhibition, the use of pig organs transgenic for human complement regulatory proteins, and conditioning regimens aimed at inducing a state of tolerance or specific immunologic hyporesponsiveness. The greatest success to date has been obtained with methods that inhibit complement-mediated injury, either by the administration of cobra venom factor or soluble complement receptor I to the recipient (with organ survival up to 6 weeks) or by the use of donor organs transgenic for human decay-accelerating factor (with organ survival up to 2 months). The future of xenotransplantation may lie in the judicious combination of current approaches. PMID- 9753332 TI - Cytotoxic lymphocyte gene expression in peripheral blood leukocytes correlates with rejecting renal allografts. AB - BACKGROUND: We have shown previously that heightened expression of the cytotoxic lymphocyte (CL) effector genes perforin (P), granzyme B (GB), and Fas ligand (FasL), is closely correlated with acute allograft rejection, particularly when two or more target genes are up-regulated. METHODS: We used quantitative reverse transcription-polymerase chain reaction to analyze CL gene expression from peripheral blood leukocytes (PBLs) and renal allograft biopsies in 31 paired samples of PBLs and renal tissue from 25 renal allograft recipients. Our aims were (1) to determine whether the expression of CL gene expression in PBLs correlates with expression of these genes in renal allograft biopsy tissue and (2) to determine whether CL gene expression in PBLs correlates with the histological diagnosis. RESULTS: Coordinate gene expression in PBLs and acutely rejecting allografts was found in 9/11 (82%) for P, 07/11 (64%) for GB, and 10/11 (91%) for FasL. Coordinate absence was found in 15/20 (75%) for P, 17/20 (85%) for GB, and 16/20 (80%) for FasL in nonrejecting allografts. Furthermore, up regulation of any two genes in PBLs correlated with pathological diagnosis of rejection with excellent positive (100%) and negative (95%) predictive values. CONCLUSION: Coordinate CL gene expression in PBLs and the allograft is usually detected. CL gene expression in PBLs is closely associated with a pathologic diagnosis of rejection. CL gene expression in PBLs may serve as a noninvasive method of monitoring for renal allograft rejection. PMID- 9753333 TI - Alterations in mRNA for inducible and endothelial nitric oxide synthase and plasma nitric oxide with rejection and/or infection of allotransplanted lungs. AB - BACKGROUND: Experiments were designed to determine expression of type II (iNOS) and type III (ecNOS) nitric oxide synthase in lung parenchyma and systemic endothelial cells with rejection and/or infection of single lung allografts. METHODS: After single lung allotransplantation, dogs were maintained on standard triple immunosuppressive therapy for 5 days and then placed into one of three groups. Group I (n=4) was maintained on immunosuppressants, group II (n=7) immunosuppression was withdrawn to allow acute rejection of the allograft, and group III (n=6) infection was induced by bronchoscopic inoculation of Escherichia coli. RESULTS: At postoperative days 7-9, no histological evidence of rejection or infection was observed in transplanted lungs of group I. In lungs of group II, rejection ranged from mild to severe; in lungs of group III, infection was severe. Some animals had both rejection and infection (n=8) and were studied separately. Plasma levels of nitric oxide increased comparably with rejection and/or infection compared to preoperative values. Expression of mRNA for ecNOS decreased significantly in lung parenchyma but not in aortic endothelial cells from dogs of groups II and III. However, expression of mRNA for iNOS increased with both rejection and/or infection in both lung parenchyma and aortic endothelial cells. CONCLUSIONS: iNOS is induced locally within the graft and systemically in aortic endothelial cells with rejection and/or infection of lung allografts. Plasma levels of nitric oxide are elevated with both rejection and infection and may not be useful in the differential diagnosis of these processes after lung transplantation. PMID- 9753334 TI - Differential in vivo recovery of sinusoidal endothelial cells, hepatocytes, and Kupffer cells after cold preservation and liver transplantation in rats. AB - BACKGROUND: The injury resulting from cold preservation/reperfusion primarily affects sinusoidal endothelial cells, while hepatocytes are thought to be less vulnerable; morphological changes and increased cytokine release suggest that Kupffer cells are activated. We evaluated the extent of functional damage to the different cell types in the liver after cold preservation and transplantation. Additionally, we analyzed in vivo the patterns of functional recovery of all three cell types over the first week after transplantation in Lewis rats. METHODS: We evaluated the in vivo uptake of hyaluronic acid, indocyanine green, and radio-labeled sulphur colloid to assess the function of sinusoidal endothelial cells, hepatocytes, and Kupffer cells, respectively. Measurements were performed immediately after transplantation using syngeneic grafts preserved in University of Wisconsin solution for different periods. Functional recovery was monitored in animals receiving grafts preserved for 24 hr over the first postoperative week. RESULTS: We found that hepatocyte were less affected compared with the profoundly damaged endothelial cells. The phagocytic ability of Kupffer cells was, however, also seriously compromised, which suggests a selective down regulation. Functional recovery occurs in a differential manner during the first postoperative week starting with hepatocytes followed by sinusoidal endothelial cells. Phagocytic function further deteriorates after transplantation before showing improvement. CONCLUSIONS: In viable liver grafts, all cell types recover from preservation/reperfusion injury by the end of the first week after transplantation. The differential time courses of the recovery suggest that successful sinusoidal endothelial cell recovery may depend upon prior hepatocyte regeneration and may involve a paracrine interaction, via cytokines and growth factors. PMID- 9753335 TI - Beneficial effects of FK409, a novel nitric oxide donor, on reperfusion injury of rat liver. AB - BACKGROUND: Nitric oxide (NO) seems to play an important role in modulating tissue injury during reperfusion of the liver. In this study, we have evaluated and compared the effects of FK409 (FK), a potent spontaneous NO releaser, and L arginine in ischemia-reperfusion injury of the rat liver. METHODS: Male Sprague Dawley rats underwent 90 min of hepatic ischemia followed by reperfusion. FK or L arginine was used (intravenously) in two different doses for each drug (group I, 3.2 mg/kg FK; group II, 1.6 mg/kg FK; group IV, 100 mg/kg L-arginine; and group V, 300 mg/kg L-arginine). Saline was used in control animals (group III). Hepatic enzyme status, microcirculation, serum nitrite (NO2-) and nitrate (NO3-) and tissue injury score were evaluated at predetermined times. RESULTS: Serum NO2 /NO3- was elevated immediately by FK treatment dose-dependently but not by L arginine. However, L-arginine caused late (6-24 hr) elevation of the NO metabolites dose-dependently. The elevation of serum aspartate aminotransferase and alanine aminotransferase was suppressed and hepatic microcirculation was improved in the FK-treated groups dose-dependently. L-Arginine also improved the microcirculation, but hepatic enzymes at 24 hr of reperfusion were significantly higher in group V than in the control group. These findings were well reflected by the extent of tissue injury in respective groups. CONCLUSION: FK treatment in the immediate reperfusion period improves hepatic microcirculation and confers a significant protective effect on hepatic ischemia-reperfusion injury in the rat. PMID- 9753336 TI - Donor blood monocytes but not T or B cells facilitate long-term allograft survival after total lymphoid irradiation. AB - BACKGROUND: Previous studies showed that a combination of posttransplant total lymphoid irradiation (TLI), rabbit antithymocyte globulin (ATG), and a single donor blood transfusion induced tolerance to ACI heart allografts in Lewis rats. All three modalities were required to achieve tolerance. The objective of the current study was to determine the subset(s) of cells in the donor blood that facilitated long-term allograft survival. METHODS: Lewis hosts received TLI, ATG, and donor cell infusion after heart transplantation. Graft survival, mixed leukocyte reaction (MLR), and intragraft cytokine mRNA were studied. RESULTS: The intravenous injection of 25 x 10(6) ACI peripheral blood mononuclear cells (PBMC) significantly prolonged graft survival as compared with that of Lewis hosts given TLI and ATG alone. Injection of highly enriched blood T cells or splenic B cells adjusted for the number contained in 25 x 10(6) PBMC failed to induce significant graft prolongation. Unexpectedly, depletion of monocytes (CD11b+ cells) from PBMC resulted in the loss of graft prolongation activity. Enriched populations of monocytes obtained by plastic adherence were more efficient in prolonging graft survival than PBMC on a per cell basis. Hosts with long-term grafts (>100-day survival) showed evidence of immune deviation, because the MLR to ACI stimulator cells was vigorous, but secretion of interferon-gamma in the MLR was markedly reduced. In situ hybridization studies of long-term grafts showed markedly reduced levels of interferon-gamma mRNA as compared with rejecting grafts. CONCLUSION: Infusion of donor monocytes facilitated graft prolongation via immune deviation. PMID- 9753337 TI - Infectious complications occurring in liver transplant recipients receiving mycophenolate mofetil. AB - BACKGROUND: Mycophenolate mofetil (MMF) is a new immunosuppressive agent that is gaining widespread use in solid organ transplantation recipients. A comprehensive assessment of infectious complications after its use after liver transplantation has never been assessed. METHODS: Bacterial, fungal, and viral infections occurring after transplantation were compared for a cohort of consecutive liver transplant recipients who received MMF (because of suspected tacrolimus-related nephrotoxicity or neurotoxicity) and a cohort who did not receive the drug. All patients received a tacrolimus-based primary immunosuppressive protocol. RESULTS: Biopsy-proven acute rejection episodes within the first 6 months after transplant occurred in 6% of MMF-treated patients but in 30% of those who did not receive MMF (P=0.07). No significant differences were found in occurrence of cytomegalovirus infection or disease, Pneumocystis carinii, Aspergillus, or other fungal infection and hepatitis C virus recurrence between MMF-treated and untreated patients. Bacterial infections were more common in MMF-treated patients, but this cohort had a prolonged intensive care unit stay compared with patients who did not receive MMF. None of the MMF-treated patients with bacterial infection had leukopenia. CONCLUSIONS: MMF use does not appear to be associated with an significantly increased risk of infection occurring after liver transplantation and is associated with fewer episodes of acute rejection. PMID- 9753338 TI - Does intraoperative hepatic artery flow predict arterial complications after liver transplantation? AB - BACKGROUND: Little is known about the value of intraoperative hepatic artery (HA) flow measurement on the development of HA complications in orthotopic liver transplantation (OLT). We undertook this study to see whether assessing HA flow at the OLT helps predict posttransplant HA complications (HA thrombosis or stenosis). METHODS: Four hundred and eleven consecutive OLT in 367 adult patients who received grafts between November 1992 and August 1995 were reviewed. Of these, 259 grafts in 255 patients with at least 1 year of follow-up and with complete data were studied. HA flow, portal vein flow, percentage of cardiac index going to HA (HA/CI), HA flow per 100 g of liver tissue, mean arterial pressure, central venous pressure, and CI were analyzed. Preservation injury was assessed by posttransplant alanine aminotransferase and aspartate aminotransferase levels. RESULTS: Thirty-four patients with 35 grafts developed HA thrombosis or stenosis during a median follow-up time of 29 months. HA complications occurring within the first 100 days of OLT were classified as early complications. HA flow at the time of surgery and percentage of CI going to the liver were found to be significant variables in early HA complications. Hepatic hemodynamics were not different in the late HA complication group compared to the control. Systemic hemodynamics and posttransplant alanine amino-transferase and aspartate aminotransferase levels were similar in all three groups. Logistic regression analysis showed that patients with HA flows less than 400 ml/min were more than 5 times as likely to develop HA complications (risk ratio 5.1). CONCLUSIONS: HA flow measurement should be obtained at the time of OLT and may help to predict early but not late posttransplant HA complications. Patients with HA flows less than 400 ml/min or HA/CI values of less than 7% may carry a higher risk for HA stenosis or thrombosis and may need close surveillance to detect such problems. PMID- 9753339 TI - Gender matching and outcome after pediatric liver transplantation. AB - BACKGROUND: Gender is not a selection criterion for orthotopic liver transplantation (OLT), and reports in adults have shown a less favorable outcome for male recipients of female organs; the only pediatric study did not support this finding. The aim of the present study was to assess the impact of donor and recipient gender on graft and patient survival rates after pediatric OLT. METHODS: We have reviewed retrospectively 137 children (male=63; median age: 3.4 years; range: 14 days to 15 years) undergoing primary OLT from January 1991 to June 1996. These children were divided into donor-recipient gender match (M; n=64) and nonmatch (NM; n=73) groups and then classified into female to female (FF; n=30), female to male (FM; n=29), male to female (MF; n=44), and male to male (MM; n=34) subgroups. RESULTS: The M group had better graft and patient survival rates at both 1- and 5-year follow-up compared with the NM group (P<0.01). Graft and patient survival rates were different among gender subgroups (P<0.04). Graft and patient survival rates in the FM group were poorer than in the MM subgroup at both 1 and 5 years (P<0.03, P<0.01). The FM group had a higher incidence of early complications than the MM (P<0.01) group, with 50% and 33% of graft losses, respectively, related to the complications. To minimize the influence of hormonal factors, we have analyzed separately the patients younger than 12 and 10 years who had similar findings. CONCLUSION: Graft and patient survival rates after pediatric OLT are worse in gender mismatch groups, particularly for male recipients of female organs. Early complications play a role in the decreased survival rates. PMID- 9753340 TI - Liver transplantation in the first three months of life. AB - BACKGROUND: Pediatric liver transplant recipients have traditionally been grouped according to age. Age-based classification schemes are useful in identifying clinical problems in selected age groups and also for developing solutions to these problems. Although infants in the first 3 months of life have not traditionally been considered a distinct age group, several features of these infants may distinguish them from other pediatric liver transplant recipients. METHODS: The experience with liver transplantation in infants during the first 3 months of life in three large pediatric liver transplant programs (University of Chicago, Stanford University, and UCLA) was analyzed in order to characterize this group. RESULTS: A total of 23 liver transplants were performed at these three centers in children younger than 3 months of age. This group of patients comprised approximately 37% of the U.S. experience between 1988 and 1994 according to United Network for Organ Sharing statistics. Age distribution at the time of transplantation included the following: <1 month, 28%; 1-2 months, 35%; and 2-3 months, 36%. Median age at the time of transplantation was 37 days (range, 7-90 days), and mean age was 57+/-30 days. Mean weight at the time of transplantation was 3.8+/-1.0 kg. Etiology of liver disease included idiopathic hepatitis, 52%; iron storage disease, 17%; and other causes, 31%. Types of liver allografts used included cadaveric, 85% (reduced size, 60%, and full-size, 25%); living donor, 15%; ABO-identical, 65%; and ABO-compatible, 35%. Actuarial patient and graft survival rates were 60% and 60% at 1 year and 60% and 42% at 2 years, respectively. Median follow-up was 1.5 years. Rejection occurred in 42% of patients, with a median time to first rejection of 13 days. Of these patients, 28% required steroids only and 14% required OKT3. Three patients (14%) were retransplanted at a median time to retransplantation of 1.6 years. Vascular thrombosis occurred in three patients (14%). CONCLUSIONS: Liver transplantation performed in infants younger than 3 months of age (1) provides acceptable short- and long-term patient and graft survival, (2) is associated with significant rates of rejection, and (3) is not associated with excessive rates of vascular thrombosis. The etiology of end-stage liver disease occurring in the first 3 months of life is distinct from that in other pediatric liver transplant recipient age groups. These infants should be referred promptly for liver transplantation as reasonable survival can be expected. PMID- 9753341 TI - Clinical and metabolic effects of different parenteral nutrition regimens in patients undergoing allogeneic bone marrow transplantation. AB - BACKGROUND: Nutrients may interfere with physiological and pathophysiologic mechanisms. The present study was aimed at evaluating whether the differences in the quality of energy substrates administered with total parenteral nutrition (TPN) after cytoreductive therapy may influence the clinical outcome of patients undergoing bone marrow transplantation (BMT). METHODS: Sixty-six consecutive allogeneic BMT patients with hematologic malignancies were randomized to receive either a glucose-based (100% glucose) or a lipid-based (80% lipid, using an omega 6 long-chain triacylglycerol emulsion + 20% glucose) TPN, providing 146.3 kJ/kg body weight, 1.4 g of protein/kg of body weight, administered from day +1 to day +15 after BMT. Time to engraftment (EGT), incidence of sepsis and metabolic complications (hyperglycemia and hypertriglyceridemia), incidence of acute graft versus-host-disease (A-GVHD) and relapse, survival at 18 months, incidence of deaths for A-GVHD and relapse were evaluated. RESULTS: Six patients dropped out before completing the study period. Thirty-one patients in the glucose-based TPN group and 29 patients in the lipid-based TPN group were evaluated. The incidence of hyperglycemia was significantly lower in the lipid-based TPN group than in the glucose-based TPN group (3.4% vs. 32%, respectively; P=0.004). Five patients in the glucose group and none in the lipid group died for A-GVHD (P<0.05). Survival at 18 months tended to be higher in the lipid group than in the glucose group (62% vs. 42%, P=NS). Rate of bone-marrow EGT, time to EGT, incidence of sepsis and fungal infections during TPN, incidence of A-GVHD, and rate of relapse at 18 months were not different in the two groups. CONCLUSIONS: The results obtained suggest that the use of lipid-based TPN after allogeneic BMT is associated with lower incidence of lethal A-GVHD and hyperglycemia, without negatively affecting the EGT of infused cells. Intravenously administered lipids might have influenced the severity of A-GVHD likely via modulation of immune response and synthesis of cytokines, prostaglandins, and leukotrienes that participate in the pathogenesis of graft-versus-host disease. PMID- 9753342 TI - Reverse seroconversion of hepatitis B after allogeneic bone marrow transplantation: a retrospective study of 37 patients with pretransplant anti-HBs and anti-HBc. AB - BACKGROUND: Reverse seroconversion to hepatitis B virus (HBV), i.e., HBV reactivation in patients with pretransplant antibodies to hepatitis B surface antigen (anti-HBs) and to hepatitis B core antigen (anti-HBc), is rarely re ported after allogeneic bone marrow transplantation. METHODS: To determine this risk, we studied clinical outcome and serological changes in 37 patients with pretransplant anti-HBs and anti-HBc. RESULTS: In 33 cases, no change in HBV markers was observed in the posttransplant period. In four cases, anti-HBs and anti-HBc were lost, and hepatitis B surface antigen, hepatitis B e antigen, and HBV DNA emerged together with acute hepatitis, after cessation of immunosuppression. The actuarial risk of reactivation in the 37 patients was 20.5% (median follow-up 20 months). No reactivation occurred in patients with anti-HBs-positive donors. CONCLUSION: Although few cases of postallogeneic bone marrow transplantation reverse seroconversion to HBV have been reported, this study demonstrates that the actuarial risk is relatively high and suggests that donor vaccination might be proposed prophylactically or that HBs-specific immunoglobulin infusions might be warranted. PMID- 9753343 TI - Low incidence of acute graft-versus-host disease, using unrelated HLA-A-, HLA-B-, and HLA-DR-compatible donors and conditioning, including anti-T-cell antibodies. AB - BACKGROUND: Using unrelated bone marrow, there is an increased risk of graft versus-host disease (GVHD). METHODS: HLA-A-, HLA-B-, and HLA-DR-compatible unrelated bone marrow was given to 132 patients. The diagnoses included chronic myeloid leukemia (n=43), acute lymphoblastic leukemia (n=29), acute myeloid leukemia (n=27), myelodysplastic syndrome (n=4), lymphoma (n=3), myeloma (n=1), myelofibrosis (n=1), severe aplastic anemia (n=12), and metabolic disorders (n=12). The median age was 25 years (range 1-55 years). HLA class I was typed serologically, and class II was typed by polymerase chain reaction using sequence specific primer pairs. Immunosuppression consisted of antithymocyte globulin or OKT3 for 5 days before transplantation and methotrexate combined with cyclosporine. RESULTS: Engraftment was seen in 127 of 132 patients (96%). Bacteremia occurred in 47%, cytomegalovirus (CMV) infection in 49%, and CMV disease in 8%. The cumulative incidences of acute GVHD > or = grade II and of chronic GVHD were 23% and 50%, respectively. The 5-year transplant-related mortality rate was 39%. The overall 5-year patient survival rate was 49%; in patients with metabolic disorders and severe aplastic anemia, it was 61% and 48%, respectively. The disease-free survival rate was 47% in patients with hematological malignancies in first remission or first chronic phase and 38% in patients with more advanced disease (P=0.04). Acute GVHD was associated with early engraftment of white blood count (P=0.02). Poor outcome in multivariate analysis was associated with acute myeloid leukemia (P=0.01) and CMV disease (P=0.04). CONCLUSION: Using HLA-A-, HLA-B-, and HLA-DR-compatible unrelated bone marrow and immunosuppression with antithymocyte globulin, methotrexate, and cyclosporine, the probability of GVHD was low and survival was favorable. PMID- 9753344 TI - Polymorphism in the human anti-pig natural antibody repertoire: implications for antigen-specific immunoadsorption. AB - BACKGROUND: Anti-Galalpha1-3Gal antibodies cause hyperacute rejection (HAR) in pig-to-primate xenotransplantation. Long-term graft survival has not been achieved despite abrogation of HAR using transgenic pigs. IgG and IgM anti Galalpha1-3Gal also play a role in the events following abrogation of HAR. Characterizing these antibodies and developing a system for their removal is therefore crucial to future success in xenotransplantation. METHODS AND RESULTS: We have developed a neoglycoprotein enzyme-linked immunosorbent assay to probe the precise antigenic requirements for the binding of anti-Galalpha1-3Gal and have analyzed 77 normal sera. Sixty-six percent of individuals have IgG that recognizes the Galalpha1-3Gal di-, tri-, and pentasaccharides (D, T, and P, respectively), termed DTP phenotype. The frequency of other phenotypes was - -P, 13%; -TP, 12%; D-P, 8%; and DT-, 1%. The IgG subclasses found were IgG2 (95%), IgG3 (34%), IgG1 (31%), and IgG4 (17%). IgM in 91% of individuals recognized all three antigens. Further antibody heterogeneity was demonstrated when immunoadsorbents derived from Galalpha1-3Galbeta1-4GlcNAcbeta1-3Galbeta1-4Glc (PENTA) were tested. Galalpha1-3Galbeta1-4Glc (TRI 6) or PENTA agarose were effective for IgG removal in all individuals. For IgM removal, two deoxy derivatives were completely successful in 73% of individuals. Combining the Galalpha1-3Gal (DI) and TRI 6 agarose produced an adsorbent that completely removed anti-Galalpha1-3Gal IgG and IgM in all individuals tested. CONCLUSIONS: Although the polymorphism in the anti-Galalpha1-3Gal repertoire, which we have demonstrated, represents a major obstacle to the development of an effective immunoadsorbent, the combination of DI and TRI 6 agarose appears sufficient for pig-to-human xenotransplantation. PMID- 9753345 TI - In vitro xenorecognition of adult pig pancreatic islet cells by splenocytes from nonobese diabetic or non-diabetes-prone mice. AB - BACKGROUND: In vitro studies of the nonobese diabetic (NOD) mouse prone to type 1 autoimmune diabetes were conducted in order to investigate the mechanisms possibly involved in cell-mediated rejection of adult pig islet xenografts. Mouse cellular proliferation in discordant situations was previously investigated only with stimulator lymphocytes and found to be low in intensity and due to an indirect recognition mechanism involving murine antigen-presenting cells (APC). It was also important to characterize murine anti-pig islet response. METHODS AND RESULTS: In the present study, mouse splenocytes responded to pig islet cells since primary proliferations were detected in non-diabetes-prone Balb/c (P<0.04) or NOD (P<0.001) mice. Moreover, NOD mice displayed a higher (P<0.003) splenocyte response to pig islet cells (stimulation index: 5.8+/-0.7) than did Balb/c mice (stimulation index: 2.3+/-0.3), whereas responses to pig stimulator splenocytes were similar in both strains. The proliferation of NOD splenocytes to pig islet cells was lower (P<0.0001) than the allogeneic response to Balb/c islet cells but similar to syngeneic proliferation to NOD islet cells. In both NOD and Balb/c mice, splenocyte proliferation to pig islet cells was abolished (P<0.01) when CD4+ cells were blocked with antibodies, whereas the blocking of CD8+ cells had a nonsignificant effect. The main T-splenocyte subsets involved were restricted to mouse MHC class II molecules as they did not proliferate in the presence of monoclonal antibodies directed at I-A molecules. NOD and Balb/c splenocyte proliferation to pig islet cells was abolished after removal of plastic-adherent APC, which indicates that the major activation pathway was indirect. Purified CD4+ or CD8- cells alone did not proliferate in response to pig islet cells but recovered a proliferative ability when mixed with APC. CD4- cells, alone or in the presence of APC, were not capable of responding to pig islet cells. Both Th1 and Th2 splenocytes were involved in response to pig islet cells since interferon gamma (IFN-gamma) and interleukin (IL-)-4 production increased significantly (300 fold and 11-fold, respectively, P<0.02 for both), whereas the increase in IL-10 production was much lower (only 1.5-fold). The IFN-gamma/IL-4 and IFN-gamma/IL-10 ratios stimulated by pig islet cells were not different with NOD and Balb/c splenocytes. CONCLUSION: In conclusion, mouse cell-mediated reaction against xenogeneic adult pig islet cells mainly involves class II-restricted CD4+ T lymphocytes of Th1 and Th2 subtypes, with an indirect pathway for the recognition. Although of low intensity, this cell-mediated reaction constitutes an obstacle to pig islet engraftment in the mouse, although one not necessarily more insurmountable than alloreactivity. The peculiarity of NOD mouse splenocytes, in terms of proliferation against pig islets, suggests that the study of islet xenograft rejection should take the immunogenetic context of diabetes into account, in which case the use of non-diabetes-prone mice has its limitations. PMID- 9753346 TI - Contrasting outcomes of donor-specific blood transfusion: effectiveness against cell-mediated but not antibody-mediated rejection. AB - BACKGROUND: Giving recipients a prior donor-specific blood transfusion (DST) is effective in prolonging organ allograft survival in some inbred strains but not in others. The present investigation analyzed two such contrasting strains of rats in an attempt to define the basis for this variation. METHODS AND RESULTS: The survival of fully mismatched Dark Agouti (RT1a) cardiac allografts was significantly prolonged (from 7 to 44 days, median survival times) in PVG (RT1c) rats given a prior (-14 day) DST, whereas it shortened survival in the high responder PVG-RT1u strain. Injecting PVG recipients with blood from strains bearing defined differences indicated that each disparity contributed to the increased survival time in an incremental way: blood and heart matched at the MHC class I (A) and/or class II (B/D) loci had a major influence on survival; class I like (C) and non-MHC antigens made only minor contributions. MHC disparities had contrasting effects in RT1u rats. Blood transfusions from Dark Agouti or PVG-R8 (AaB/DuCu) rats induced accelerated rejection and anti-Aa alloantibody formation; transfusing PVG-R23 (AuB/DaCa) blood, a class II and class I-like difference, induced indefinite R23 heart allograft survival. Although produced in high titer, anti-class II antibody was not able to induce rejection in RT1u rats. Specific anti-Aa alloantibody was able, after passive transfer, to destroy class I disparate allografts in both RT1u nude and PVG nude recipients. However, under normal circumstances, acute rejection in the PVG strain occurred in the absence of anti-Aa antibodies, presumably by a cell-mediated mechanism. CONCLUSION: Anti class I alloantibody, when produced, seemed to override the unresponsiveness induced by DST. The results indicated that DST was effective only when rejection was induced by a cell-mediated response. The two contrasting response patterns in animals may reflect the experience of transplant patients who either benefit from DST or become sensitized instead. PMID- 9753347 TI - Alloreactive cytotoxic T cells recognize minor transplantation antigens presented by major histocompatibility complex class Ib molecules. AB - BACKGROUND: Cytotoxic T lymphocytes (CTLs) contribute to the rejection of transplanted tissues through two pathways: first, by direct recognition of foreign graft major histocompatibility complex (MHC) class I molecules; and second, by recognition of foreign graft-derived peptides presented by classical MHC class Ia molecules that are matched between graft and donor. However, a number of observations suggest that additional categories of CTL recognition patterns may exist, but they remain to be defined molecularly. METHODS: Previous studies showed that the murine nonclassical MHC molecule H2 M3 may be involved in allorecognition. We investigated whether other members of nonclassical MHC class Ib, namely Qa1 and Qa2, may be recognized. Alloreactive CTLs were generated from mice mismatched for non-MHC and/or MHC genetic backgrounds and tested using various target cells, including cells transfected with Qa1 or Qa2. Furthermore, candidate peptides were synthesized and used to generate CTLs specific for peptide presented by Qa1 or Qa2. RESULTS: The experiments demonstrate that allogeneic and xenogeneic peptides were recognized by CTLs when presented on shared nonclassical MHC class Ib Qa1 and Qa2 molecules. CONCLUSIONS: The results confirm that MHC class Ib molecules present peptides to CTLs. This potentially important alloreactivity pathway may be functional between most individuals because sharing of MHC class Ib alleles is frequent. PMID- 9753348 TI - Acute limb ischemia after transplantation in a patient with persistent sciatic artery. AB - BACKGROUND: Improved surgical techniques have contributed to a better outcome in kidney transplantation, and graft failure secondary to technical complications is unusual. METHODS: We describe a surgical complication secondary to a rare and unexpected vascular anomaly in a pediatric renal transplant that resulted in considerable morbidity. RESULTS: Our patient had a persistent sciatic artery as a dominant source of blood supply to the lower limb. Ligation of this vessel during anastomosis with the transplant kidney vasculature led to severe ischemic damage to the leg. CONCLUSION: This vascular malformation, if undetected, can lead to serious complications. We present a review of the literature regarding this malformation and its management with regard to renal transplantation. PMID- 9753349 TI - Bacterial endocarditis associated with crescentic glomerulonephritis in a kidney transplant patient: first case report. AB - BACKGROUND: Endocarditis-induced crescentic glomerulonephritis is a well described complication in nontransplant patients. Its occurrence in transplant patients has not been reported to date. METHODS: A 50-year-old man who had received a renal allograft 13 years before and been treated with prednisone, 10 mg/day, was admitted for progressive renal failure, purpura, edema of the lower limbs, and fever. RESULTS: Blood cultures isolated Streptococcus bovis and cardiac ultrasound examination revealed a 23-mm-large vegetation on the mitral valve. His plasma creatinine level was 478 micromol/L and his proteinuria was 5.5 g/day. A renal biopsy showed diffuse crescentic glomerulonephritis. Long-term antibiotic treatment and three methylprednisolone pulses were effective in treating the endocarditis and glomerulonephritis. CONCLUSION: Endocarditis induced glomerulonephritis is an immune-mediated disease that can also occur on a renal allograft. It is likely that a low daily dose of immunosuppressive treatment may have been a facilitating factor. PMID- 9753350 TI - Advanced donor-origin melanoma in a renal transplant recipient: immunotherapy, cure, and retransplantation. AB - BACKGROUND: A kidney transplant recipient inadvertently contracted donor-origin melanoma, which was found to be very advanced at presentation. Withdrawal of immunosuppression failed to induce rejection, and interferon-alpha was required. When florid allograft rejection was in progress, the allograft was removed, before it was recognized that the transplanted melanoma was not being simultaneously rejected. METHODS: Subsequent immunotherapy was required, which largely recapitulated treatment of recognized value in autologous melanoma and included interferon-alpha, use of cultured melanoma cells as tumor vaccine, pooled allogeneic cell vaccination, and adoptive immunotherapy using lymphokine activated killer cells. RESULTS: Prolonged immunotherapy eradicated the widespread malignancy, and the patient went on to a successful second renal transplant, with follow-up of over 24 months. CONCLUSIONS: This unique case demonstrates the successful cure of advanced transplanted melanoma through the use of immunotherapy, which did not require sophisticated tumor vaccine technology, and successful retransplantation. PMID- 9753351 TI - Concomitant caudate lobe resection as an option for donor hepatectomy in adult living related liver transplantation. AB - In this article, we describe a successful adult living related partial liver transplantation (LRLT) using the left lobe with the left-side caudate lobe (the Spiegel lobe and the left side of the paracaval portion). The size of the donor's left lobe was 29% of the recipient's standard liver volume and did not seem to meet our criteria for adult-to-adult LRLT. However, the donor had a thick left side caudate lobe. The estimated volume of the left lobe with the left-side caudate lobe was 32%, which met our criteria for the adult recipient. The recipient's CT scan on day 87 after transplantation showed the preserved blood flow and no biliary congestion in the left-side caudate lobe, which suggests maintenance of lobe function. This procedure may be an option for adult-to-adult LRLT in which the donor has a thick left-side caudate lobe. PMID- 9753352 TI - Detection of hepatitis C virus replication in peripheral blood mononuclear cells after orthotopic liver transplantation. AB - BACKGROUND: The presence of hepatitis C virus (HCV) replication in peripheral blood mononuclear cells (PBMCs) remains controversial. We determined the presence of the negative HCV RNA strand in PBMCs from a group of HCV-positive patients before and after liver transplantation. METHODS: Nine patients receiving orthotopic liver transplantation for end-stage HCV-related liver disease were studied. PBMCs were collected on the day of transplantation and 1 month later. The negative HCV RNA strand was detected by highly strand-specific Tth-based reverse transcriptase polymerase chain reaction. RESULTS: All nine patients were positive for the presence of the HCV RNA-positive strand in serum and PBMCs both before and after transplantation. The presence of the negative HCV RNA strand was documented in three PBMC samples after transplantation but in none of the samples collected before transplantation. CONCLUSION: Our results suggest that under circumstances of impaired immunity associated with pharmacological immunosuppression, HCV may be lymphotropic in vivo. PMID- 9753353 TI - Squamous cell carcinomas after allogeneic bone marrow transplantation for aplastic anemia: further evidence of a multistep process. AB - BACKGROUND: Secondary solid tumors are rare events occurring in patients who underwent allogeneic marrow transplantation for aplastic anemia and Fanconi's anemia. Human herpes virus 8 (HHV8), Epstein-Barr virus (EBV), and human papillomaviruses (HPV) sequences have been found in squamous cell carcinoma (SCC) occurring in organ transplant recipients. The tumor suppressor gene p53 has been strongly linked to the occurrence of SCC in the nonimmunocompromised population. PATIENTS AND METHODS: In eight patients with SCC, we searched for HHV8, EBV, varicella zoster virus, adenovirus, and HPV sequences from DNA extracted from selected areas of SCC. We also looked for p53 expression in those specimens as well as the presence of anti-p53 antibodies in the serum of these patients at the onset of SCC. RESULTS: In one patient, we found the presence of both HHV8 and EBV sequences, and in another patient we found HPV16 sequences. All five tumors that could be studied disclosed evidence of p53 accumulation, but none of the eight patients had anti-p53 antibodies in the sera. CONCLUSION: SCC developing in marrow transplant recipients seems to occur via a multistep process. Genetic predisposition may be present, as in patients with Fanconi's anemia. Transplantation-related factors, such as irradiation and chronic graft-versus host disease, also have a role. In this article, we add two more potent risk factors: p53 alteration(s) and in some cases the presence of oncogenic viruses. PMID- 9753354 TI - Mononuclear phagocyte populations in the transplanted human lung. AB - BACKGROUND: Dendritic cells (DC) are essential for the development of alloreactivity, however, little has been published regarding the distribution and phenotype of these and related mononuclear cells in human lung transplantation. METHODS: Lung frozen sections were examined for the presence of CD1a+ DC and for mononuclear cells and alveolar macrophages expressing CD11b and CD68. The effects of transplantation and immunosuppression were assessed by comparison of normal transplant transbronchial biopsy specimens to specimens from unused donor lungs; the normal transbronchial biopsy specimens also were compared with those showing rejection or obliterative bronchiolitis. RESULTS: All biopsy specimens, including those with obliterative bronchiolitis, showed a marked depletion of CD1a+ DC in lung allografts. This has not been described previously. In addition, transplantation and immunosuppression reduced alveolar macrophage coexpression of CD68 and CD11b, and this was reversed in acute rejection. CONCLUSION: The roles of pulmonary DC and other mononuclear phagocyte subpopulations need to be further defined, and data from animal models of lung transplantation should be interpreted with caution. PMID- 9753355 TI - Successful kidney pancreas transplantation from donor with methanol intoxication. PMID- 9753356 TI - Using the Internet--an example. PMID- 9753357 TI - SEA's pipe explorer. Science and Engineering Associates. PMID- 9753358 TI - From chimney sweeps to astronauts: cancer risks in the work place: the 1998 Lauriston Taylor lecture. AB - Percival Pott, in 1775, was the first to note an association between overt cancer and a carcinogen in the work place when he astutely observed an elevated incidence of scrotal cancer in small boys who assisted chimney sweeps. In their "workplace" astronauts and crew of high altitude jet-liners are exposed, not only to low linear energy transfer (LET) radiation but also to HZE (high energy + high atomic number) particles and to neutrons-for which no human epidemiological data exist. The current system of radiation protection is based on risk estimates from low LET radiations, delivered in large doses and at high dose-rate, coupled with the assumption of a linear no-threshold model. In extrapolating to low doses and dose-rates, and to high LET radiations, it would be helpful if the mechanisms of radiation carcinogenesis were known. Unfortunately that is not the case, though progress has been made toward that end. Many human leukemias and lymphomas appear to be due to specific chromosomal translocations, while solid tumors usually involve multiple mutations in oncogenes, deletions in suppressor genes, and/or chromosomal rearrangements. Genomic instability and immortality are hallmarks of cancer and it is attractive to hypothesize that this is due to a mutation in a gene or genes responsible for the stability of the genome. Examples abound of a small DNA change inactivating a gene and leading to major biological consequences. This could result from a single particle, especially a HZE particle, or a recoil proton from the absorption of a neutron. In this context the assumption of a threshold is hazardous, and the linear no-threshold hypothesis still appears to be prudent and conservative. PMID- 9753359 TI - Informing the public about radiation--the messenger and the message: 1997 G. William Morgan lecture. AB - I am greatly honored to be invited by the Presidents Emeritus Committee of the Health Physics Society as a G. William Morgan Lecturer for 1997. The topic of this Plenary Session on Public Information and Public Relations is very close to my heart; it was a theme for my term as President of the International Radiation Protection Association (IRPA). I met IRPA members from all of the societies affiliated to IRPA and found that they shared a common concern about the need to improve public information especially in the event of a nuclear emergency. But who should tell them and what should be the message? There is considerable agreement about the desired characteristics of the messenger in risk communication. These include credibility, openness, and the sharing of uncertainty. The profession must maintain the status and credibility of the members, it must train members in communication skills, and above all win the cooperation of other professions. There are many obstacles to radiation protection communication, and, in particular, the complex language, derived from research, should be reserved for colleagues, and our message to the public must be clarified and freed from unnecessary jargon. Communication would be more efficient and possibly cause less anxiety if people were better educated about ionizing and non-ionizing radiation. There is considerable disagreement within the profession about the content of our message to the public. Consistency in the message would be helpful although it would be wrong to expect total unanimity in research. The profession should seek the support of the international agencies and commissions to use plain and consistent language wherever possible. I will discuss the desired characteristics of the messenger, the nature of the message, and examine some of the obstacles in the path of communication using evidence from experience with IRPA and the European Union. In this paper I will suggest some action to improve radiological protection communication and will conclude with a discussion of the central role of education. Our objective is to ensure that everyone recognizes that radiation protection opens the door to the benefits of the applications of radiation in medicine and industry. PMID- 9753360 TI - Diagnosis of acute localized irradiation lesions: review of the French experimental experience. AB - In the last 50 years several radiation accidents occurred in which industrial radiographers and others suffered severe radiation injuries from inadvertent contact with radiation sources. Such accidents involving acute localized injuries are characterized by a severe initial reaction progressing through erythema to skin necrosis with a spontaneous resolution of the lesion over a 2-mo period for the lower doses. However, the early symptoms observed on the skin give no indication as to the in-depth pathology, and cutaneous and muscular radionecrosis started generally from early epithelial, microvascular, and vascular lesions and from delayed muscular and connective tissue lesions. In a case of acute localized irradiation, different biophysical techniques are able to give real responses in biological dosimetry. More numerous are the methods, especially imaging methods, that make it possible for the clinician to evaluate the extent of the early injuries and to manage the medical intervention. We have developed animal experimental models of acute localized irradiation: overexposure to the gamma rays of a 192Ir industrial radiographic collimated source (in the pig and the rabbit) and overexposure to the beta rays of a 90Sr-90Y collimated source (in the pig). In these experimental models, most of the imaging techniques used in clinical practice, as infra-red thermography, microwave thermography, cutaneous and tissular vascular scintigraphy (beta or gamma emitters), cutaneous blood flow measurements by cutaneous laser Doppler, x ray computed tomography, nuclear magnetic resonance imaging, and skin topography, were correlated with clinical evaluation and histopathological observations, after high doses of gamma or beta irradiations ranging from 4 to 340 Gy at the skin surface. All these techniques are not for isolated use and the present review indicates that their combination is necessary to give an improved diagnostic and prognostic picture of early and late delayed radiation damage to the skin and subcutaneous tissues. PMID- 9753361 TI - The need for better methods to determine release criteria for patients administered radioactive material. AB - In current NRC regulations, three options exist that may be used to determine release criteria for patients administered radioactive materials. Absorbed dose estimates may be based on administered activity, measured dose rate, or on patient-specific calculations. All of these methods proposed by the NRC can lead to overestimation of the dose equivalent to others due to their oversimplified nature. The primary oversimplifications are the use of a point source methodology and using the measured surface entrance dose rate to determine whole body dose. In order to show the inaccuracy of these oversimplifications for 131I, results using Monte Carlo radiation transport analysis with simplified anthropomorphic mathematical phantoms were determined. These results were then compared to actual patient measurements and the results of point source analysis. The measurement data were taken from 49 131I radioimmunotherapy patients. The point source calculations were performed using well established methodologies and using the same assumptions as in the NRC regulations for patient release criteria. Monte Carlo results were obtained by implementing two simplified 70 kg anthropomorphic phantoms and performing radiation transport simulation. The activity in the "patient" phantom was assumed to be localized in the abdominal region to correspond to the activity localization seen in the radioimmunotherapy patients who were measured. Dose equivalents per unit cumulated activities were determined for 131I using the various methods. The relationship between measured dose equivalent per unit cumulated activity and whole body dose equivalent per unit cumulated activity was also investigated using Monte Carlo analysis. The point source method as implemented by the NRC yields an estimated dose equivalent per unit cumulated activity of 1.6 x 10(-8) mSv MBq(-1) s(-1) at 1 m (2.2 x 10(-4) rem mCi(-1) h(-1) at 1 m), and the Monte Carlo based method yielded a whole body dose equivalent per unit cumulated activity in the target phantom of 6.8 x 10(-9) mSv MBq(-1) s(-1)(9.0 x 10(-5) rem mCi(-1) h(-1)) for abdominal localization of activity in the source phantom. The measurements of the radioimmunotherapy patients yielded an average result of 1.0 x 10(-8) mSv MBq(-1) s(-1) (13 x 10(-4) rem mCi(-1) h(-1)). When corrected for the difference between measured surface dose equivalent and whole body dose equivalent as determined by Monte Carlo analysis, these measurements represent a whole body dose equivalent per unit cumulated activity of about 6.2 x 10(-9) mSv MBq(-1) s(-1) (8.1 x 10(-5) rem mCi( 1) h(-1)). Based on these results, the current NRC dose-based methodology for the release of patients administered radioactive materials significantly overestimates the dose equivalent to others from 131I therapy patients. PMID- 9753362 TI - Assessment of physico-chemical and biokinetic properties of uranium peroxide hydrate UO4. AB - Comprehensive studies on the radiotoxicological risk of an intermediate compound UO4, which is not specified in ICRP Recommendations, were motivated by its increased use in the nuclear fuel cycle and the lack of information such as physico-chemical and biokinetic properties. The aim of this work was to give an experimental basis for assessing the appropriate limits on intake for workers exposed to UO4 and to provide guidance for the interpretation of personal monitoring data. Particle size measurement of the UO4 dust indicated a geometric diameter D of 0.5 microm, which corresponds to an activity median aerodynamic diameter (AMAD) of 1.1 microm. In vitro experiments conducted in culture medium showed that UO4 is a soluble compound with 66.2% dissolved in 1.9 d and 33.8% in 78 d. Results of dissolution obtained with macrophages showed a significant decrease of 50% at 1 d in terms of solubility. Biokinetic data in the rat obtained from two in vivo studies involving intratracheal instillation in rats indicated half-times in the lung of 0.5 d (96.6%) and 27 d (3.4%) for an initial lung deposit (ILD) of 195 microg, and 1.2 d (90.3%) and 38 d (9.7%) for an ILD of 7.6 microg. Absorption parameters to blood as defined in the ICRP Publication 66 human respiratory tract model were calculated with the specific software GIGAFIT and led to the rapid fraction fr (0.800 to 0.873), the rapid rate sr (0.525 to 0.928 d(-1)), and the slow rate ss (1.57 x 10(-2) to 2.42 x 10(-3) d(-1)). Effective dose coefficients by inhalation for this UO4 compound using the in vivo experimental results were calculated to be between 0.52 and 0.70 x 10(-6) Sv Bq( 1). Comparison of these values with effective dose coefficients defined in ICRP Publication 68 for workers showed that UO4 could be considered as a fast soluble compound of Type F. PMID- 9753363 TI - Self-absorption of tritium betas in metal tritide particles. AB - Inhaling metal tritide particles is a potential occupational hazard. The radiation dose to tissue from tritide particles depends on their solubility and retention in the body. In each tritide particle, a portion of the beta particles from decay of tritium is absorbed by the metal matrix and therefore cannot contribute to absorbed radiation dose to tissue. A theoretical model for estimating the self-absorption of tritium betas in spherical metal tritide particles is presented. Numerical calculations are made with this method for titanium, zirconium, and erbium particles from 0.5 to 50 microm in diameter. The tritium spectrum is divided into energy groups to facilitate estimation of the energy that escapes the particle for dose calculations. Our results show considerable absorption of beta particles and their energy, even for respirable particles smaller than 5 microm. Limited experimental data of self-absorption for titanium and zirconium tritides supported the theoretical calculation. It is concluded that the self-absorption factors should be required for counting tritide particle samples as well as for estimating absorbed radiation dose to tissue. PMID- 9753364 TI - Measurement of neutrons produced in an intermediate energy heavy ion reaction using an activation technique. AB - The fluence rate, energy, and angular distributions of neutrons produced by the reaction of 50 MeV/u 12C ions on a thick copper target around the experimental target in the physics experimental hall of Heavy Ion Research Facility of Lanzhou were studied. The neutrons were measured with threshold activation detectors. Aluminum, fluorine, carbon, aluminum, and indium activation samples were used to measure neutrons with energies greater than 7 MeV, 11 MeV, 20 MeV, 50 MeV, and thermal neutrons, respectively. The fluence rate, energy and angular distributions of neutrons, total neutron yield of 12C ions, and the emission rate in the forward direction of neutrons with energies over 11 MeV and 20 MeV were obtained. PMID- 9753365 TI - The influence of house characteristics on the effectiveness of radon remedial measures. AB - The effectiveness of remedial measures in houses with high radon levels has been tested in 943 houses in the UK. Radon levels were measured for 3 mo before and after remediation, and the results were corrected for typical seasonal variations. Householders completed questionnaires about house characteristics and remedial measures. The results were analyzed to determine the influence of house characteristics on the effectiveness of different remedial measures. Significant differences in effectiveness were found, in particular depending on the age of the house and whether the measures were installed by a major contractor, a local builder, or the householder. PMID- 9753366 TI - A survey of 222Rn concentrations in domestic water supplies of Iran. AB - As part of a national program to determine public exposure to natural radiation, 222Rn concentrations were determined in domestic water supplies, including ground and surface waters, in 23 provincial centers using a liquid scintillation counting technique. The minimum and maximum mean concentrations of 222Rn in ground water were, respectively, 7.9+/-4.5 kBq m(-3) in Sanandaj and 46.5+/-11.5 kBq m(-3) in Tehran with an overall national mean value of 21+/-8.3 kBq m(-3). The 222Rn concentrations in surface waters ranged from less than 1 to 7 kBq m(-3) with a mean value of 3.9+/-1.9 kBq m(-3). The mean concentration of 222Rn in tap water in different parts of Tehran is 3.8+/-1.1 kBq m(-3). The results are presented and discussed in this paper. PMID- 9753367 TI - Does age of the host affect growth rates of skeletal malignancies? AB - Statistical analysis of bone tumor growth rates as a function of age at initiation of radiation-induced skeletal malignancies in our animal colony indicated that the p value for an association between these parameters was <0.05, suggesting a correlation in beagle dogs. The youngest animals appeared to exhibit the most slowly growing tumors, and the trend was toward more rapidly growing tumors with increasing age. Less effective immune systems in older animals were invoked as a possible explanation of this relationship. PMID- 9753368 TI - An improved mouth-piece to prevent environmental contamination during radioaerosol inhalation procedures. AB - The literature suggests that environmental contamination is common during labeled aerosol inhalation procedures in nuclear medicine. We have tested an adherent mask to prevent environmental contamination in 70 procedures. Two groups of patients were evaluated. Group 1 (60 inhalation cases in which the mask was used) presented no environmental contamination in 95% of the procedures (means of 553 dpm and 596 dpm before and after inhalation, p > 0.05, mean of the differences before/after inhalation 6.95, SD = 21.2 dpm) and the only 3 cases in which contamination did occur concerned bearded men; Group 2 (10 inhalation cases in which the mask was not used) showed large increases of environmental radioactive levels in 70% of the procedures (means of 601 dpm and 2,756 dpm before and after inhalation, p < 0.05, mean of the differences 3,066, SD = 2,98 dpm). We conclude that such a mask is very helpful in avoiding environmental contamination during radioaerosol inhalation procedures. PMID- 9753369 TI - Natural background radiation and cancer death in Rocky Mountain states and Gulf Coast states. AB - Calculations based on data from NCRP reports show that the average level of natural background radiation (NBR) in Rocky Mountain states is 3.2 times that in Gulf Coast states. However, data from the American Cancer Society show that age adjusted overall cancer death in Gulf Coast states is actually 1.26 times higher than in Rocky Mountain states. The difference from proportionality is a factor of 4.0. This is a clear negative correlation of NBR with overall cancer death. It is also shown that, comparing 3 Rocky Mountain states and 3 Gulf Coast states, there is a strong negative correlation of estimated lung cancer mortality with natural radon levels (factors of 5.7 to 7.5). PMID- 9753371 TI - Discrepancies in guidelines for exposure limits around 300 GHz. PMID- 9753370 TI - Determination of the shielding for an unoccupied roof of a radiation therapy facility to account for adjacent buildings--modification of a computer program. AB - An addition has been made to an existing software program for calculating shielding thicknesses of barriers for megavoltage radiation therapy beams. This addition calculates the shielding thicknesses required for the roof of a single story building when adjacent, multi-level buildings are occupied. PMID- 9753372 TI - New ICNIRP guidelines for human exposure to 50 Hz E- and M-fields. PMID- 9753373 TI - Response to questions and comments on ICNIRP. PMID- 9753374 TI - Early health-related behaviours and their impact on later life chances: evidence from the US. AB - This paper uses evidence from the US to examine the impact of adolescent illegal consumption and violent behaviour on later life chances. Specifically, we look at the effect of such behaviour by young men in late adolescence on productivity and household formation 10 years on. We find that alcohol and soft drug consumption have no harmful effects on economic prospects in later life. In contrast, hard drug consumption and violent behaviour in adolescence are both associated with lower productivity even by the time the individuals are in their late twenties. These effects are substantial and affect earnings levels and earnings growth. These results are robust to the inclusion of a rich set of additional controls measuring aspects of the individuals' backgrounds. However, we find no evidence of any of these behaviours significantly affecting household formation. PMID- 9753375 TI - Participation, heterogeneity and dynamics in tobacco consumption: evidence from cohort data. AB - In this paper we look at the behaviour of households as far as participation and rate of consumption of tobacco are concerned using cohort data from the Spanish Continuous Family Expenditure Survey during the period 1985-94. We test the results, in statistical and economic terms, from several estimators on samples with different levels of aggregation and offer evidence on the different behaviour of households according to several demographic characteristics. The results suggest that the effect of legislative measures cannot be identified when participation and consumption are not separately considered. Once we do so, these measures seem to affect participation alone. PMID- 9753376 TI - Measuring hospital cost efficiency with panel data models. AB - This paper investigated the development of hospital cost efficiency and productivity in Finland in 1988-1994 using a comparative application of parametric and non-parametric panel models. Stochastic cost frontier models with a time-varying inefficiency component were used as parametric methods. As non parametric methods various DEA models were employed to calculate efficiency scores and the Malmquist productivity index. The results revealed a 3-5% annual average increase in productivity, half of which was due to improvement in cost efficiency and half due to technological change. The results by parametric and non-parametric methods compared well with respect to individual efficiency scores, time-varying efficiency and technological change. The state subsidy reform of 1993 did not seem to have any observable effects on the hospital efficiency. PMID- 9753377 TI - Unobserved heterogeneity and censoring in the demand for health care. AB - In this paper we estimate a demand for private medical services equation based on the tradition of Grossman's model of demand for health using data for a panel of Spanish households. The econometric specification accounts for the censored nature of the data, which arises from no participation and infrequency of purchases, and the existence of unobserved heterogeneity, which arises from the non-observability of health states. Our evidence suggests that ignoring these features can have a significant impact on the size, sign and significance of the model estimates. The estimates for the participation and consumption processes also suggest that the deduction of expenditures on health care currently applicable in the Spanish tax system are positively associated to income and fertility. PMID- 9753378 TI - Controlling for the endogeneity of peer substance use on adolescent alcohol and tobacco use. AB - This study examines whether the effects of peer substance use on adolescent alcohol and tobacco use are due to endogeneity of adolescents selecting their peer group. We analyzed data collected for a longitudinal analysis of a drug-use prevention programme for upper elementary school students. We used a two-step probit regression to control for the potentially endogenous explanatory variable peer substance use. Rigorous tests of endogeneity and the validity of the instrumental variables showed that controlling for the endogeneity of peer substance use to reduce bias is not worth the reduction in mean squared error in these data. Peer substance use has a positive and significant effect on adolescent substance use for both drinking and smoking. These results imply that peer influence is empirically more important than peer selection (endogeneity) in our sample of adolescents in grades 6-9. Living in a single-parent family was by far the strongest predictor of adolescent drinking and smoking. PMID- 9753379 TI - The efficient organization of blood donation. AB - This paper models the costs of collecting whole blood in the north of Scotland in order to investigate strategies whereby the annual collection target can be met at lower cost. Data on the costs of the individual sessions held in 1993-1995 are analyzed using multilevel analysis. A new technique, namely the conditioned iterative generalized least squares (CIGLS) estimator is applied. Then the feasibility of collecting increased volumes from particular panels and areas is assessed by examining which factors determine the number of blood donors at a session. Results show that fixed cost and marginal cost vary across panels but marginal cost does not vary by volume. This implies that the cost-minimizing policy is to equalize marginal costs and collect higher volume at fewer panels (those with lower fixed costs). The level of donations can be increased by increasing the number of opportunities to donate and/or increasing the average length of a session. The latter policy is shown to be more cost-effective. Multilevel analysis proves not only to be appropriate but also particularly useful. PMID- 9753380 TI - Risk adjustment and the trade-off between efficiency and risk selection: an application of the theory of fair compensation. AB - We exploit the similarity between the problem of risk adjustment with prospective reimbursement schemes in the health care sector and the problem of fair compensation analysed in the social choice literature. The starting point is the distinction between two sets of variables in the explanation of medical expenditures: those for which the insurers (or the providers) can be held responsible, and those for which they have to be compensated. Using this partitioning the objectives of cost-efficiency and no risk selection can be expressed in terms of two simple axioms. If the medical expenditure function is additively separable in the two sets of variables, there exists a natural division rule which is analogous to the standard linear risk adjustment schemes. We show how this rule should be applied if the total level of actual medical expenditures is different from the budget to be divided over the insurers (or providers) and how information from the disturbances in the regression equation can be used in an optimal way. We discuss the analogy with mixed reimbursement systems. If the medical expenditure function is not additively separable in the two sets of variables, the conflict between efficiency and risk selection is unavoidable, even if one has perfect information about that function. The theoretical results are illustrated with empirical results derived from the Belgian setting where the move towards prospective reimbursement of the mutualities has necessitated the introduction of a risk adjustment formula. PMID- 9753381 TI - Understanding medically unexplained physical symptoms: faster progress in the next century than in this? PMID- 9753382 TI - Reciprocity in basic social exchange and health: can we reconcile person-based with population-based psychosomatic research? PMID- 9753383 TI - Emotion and immunity. AB - Earlier studies have suggested that depression is associated with decreased immune function, but a recent literature review has revealed that a majority of studies reached inconsistent or conflicting conclusions. On the other hand, studies on immune function in anxiety disorders are sparse, and their findings are also inconsistent. Despite a few contradictory results, a clinical level of anxiety seems to reduce immune function, whereas a subclinical level of anxiety seems to enhance immunity. The latter may be a transient phenomenon occurring prior to the downregulation of immune function, reflecting the body's defense to a stressor. Thus, research needs to be conducted to elucidate the relationship between those hormones related to hypothalamic-pituitary-adrenal axis and a variety of immune measures at the subclinical level of anxiety. In addition, to confirm the interaction between emotion and immune function, the effectiveness of treatment with medication and psychotherapy on immunity should be investigated. PMID- 9753384 TI - Depression and social functioning in general hospital in-patients. AB - Impairment of social functioning is often associated with depression and contributes to an unfavorable course of the disease. Although it must be suspected that both social maladaptation and depression could obstruct recovery from somatic diseases, little attention has been paid to their interaction in general hospital patients. To assess social integration in depressive and psychiatrically healthy general hospital in-patients, 250 patients were studied with the Composite International Diagnostic Interview (CIDI), a structured clinical interview, and the Social Interview Schedule (SIS). From clinical interviews, it was established that 16.4% of the patients suffered from depressive disorders (ICD-10). When these patients were compared with patients without psychiatric disorder, only a tendency to social dysfunctioning with regard to social management and satisfaction with social situations was observed. But when the depressive sample was divided into three diagnostic groups (depressive episode, dysthymia, depressive adjustment disorder), significant social impairments were found in the dysthymia subsample. Family and other interpersonal problems were most prominent. When depression preceded somatic illness, a higher level of impairment was observed. The majority of dysthymia patients suffered from long-term somatic diseases, often cancer, which were preceded by depression. The results of this study single out a small group of patients who seem to be at an extensive risk of chronic psychiatric and somatic illnesses and should therefore be a focus of consultation/liaison (C/L) interventions. PMID- 9753385 TI - The course of anxiety and depression in patients undergoing coronary artery bypass graft surgery. AB - A semilongitudinal study was designed to follow-up the course of anxiety and depression in patients undergoing coronary artery bypass graft (CABG) surgery. The focus was on possible effects of gender and age on variations in both mean level and interindividual differences over time. At two timepoints before and two after surgery, 217 patients completed self-report questionnaires. Multivariate testing revealed an overall decrease in mean levels of anxiety and depression in the postoperative period but different trends for men and women. Compared with men, women reported more anxiety and depression, both pre- and postoperatively, but showed a relatively stronger decrease in the early postoperative period. Regarding variations in interindividual differences over time, multivariate testing revealed different trends of depression for men and women. Women appeared to be most homogeneous in the early days after surgery, whereas interindividual differences for men showed a stable trend. PMID- 9753386 TI - Defensiveness and essential hypertension. AB - The purpose of this study was to investigate the association between essential hypertension and defensiveness. Fifty normotensive and 74 hypertensive subjects completed the State-Trait Personality Inventory (STPI) and State-Trait Anger Expression Inventory (STAXI) to assess perceived anger and anxiety, and the Marlowe-Crowne Scale of Social Desirability as an indicator of defensiveness. Hypertensive and normotensive groups did not differ in their scores on the anger, anger expression, and anxiety scales. In contrast, Marlowe-Crowne scores were higher in the hypertensive group (18.1+/-5.5 vs. 15.4+/-5.1) (p=0.006). Stepwise logistic regression that included age, gender, BMI, and Marlowe-Crowne scores (dichotomized at 18) showed that a high Marlowe-Crowne score was associated with a relative risk of 3.63 (CI 1.49-8.83) of being hypertensive, independent of age, gender, and BMI. Anger and anxiety scores did not predict hypertensive status and did not affect the relationship between Marlowe-Crowne score and hypertensive status. We conclude that defensiveness is more closely related to essential hypertension than is self-reported anger or anxiety. Better understanding of conscious and unconscious mechanisms of defensiveness are likely to be important in clarifying the link between emotions and hypertension. PMID- 9753387 TI - Assessing adherence to dietary recommendations for hemodialysis patients: the Renal Adherence Attitudes Questionnaire (RAAQ) and the Renal Adherence Behaviour Questionnaire (RABQ). AB - This article describes preliminary investigations into the psychometric properties of two scales for hemodialysis patients (N=35): the Renal Adherence Attitudes Questionnaire (RAAQ), a 26-item scale measuring attitudes toward adherence: and the Renal Adherence Behaviour Questionnaire (RABQ), a 25-item scale measuring self-reported dietary (diet and fluid) adherence. Factor analysis of the RAAQ yielded a four-factor structure. These factors were attitudes to social restrictions, well-being, self-care/support, and acceptance. The scale demonstrated high internal and test-retest reliability. Factor analysis of the RABQ gave a five-factor structure: adherence to fluid restrictions; adherence regarding potassium and phosphate restrictions, adherence regarding self-care; adherence regarding sodium intake; and adherence in times of particular difficulty. This scale had moderately high internal reliability and high test retest reliability. Validity for the RABQ was tested with independent measures of adherence; biochemical (serum potassium, serum phosphate, and interdialytic weight gain) and dietician-rated (potassium and fluid). There was little association among the differing measures of adherence. These scales facilitate empirical evaluation of dietary adherence for hemodialysis patients. PMID- 9753388 TI - Power spectral analysis of heart rate variability in type A females during a psychomotor task. AB - This study investigated changes in autonomic nervous activities due to psychological stress in Type A females. Eight Type A and eight Type B females performed a psychomotor task for 30 minutes. Power spectral analysis of heart rate variability (HRV) was used to examine the autonomic nervous activities. Results showed the low frequency (LF) component and LF/HF ratio in Type A females increased after the onset of the task. There were no significant differences in task performance between Type A and Type B females. The subjective mental workload increased gradually in Type A females during the tasks, whereas in Type B females this parameter did not change in a consistent manner. The results suggest that the sympathetic nervous system in Type A females was more stimulated by the task and Type A females felt a greater subjective mental workload than did Type B females. PMID- 9753389 TI - Features of eating disorders in patients with irritable bowel syndrome. AB - The relationship between characteristics of irritable bowel syndrome (IBS) and eating disorders (ED) was investigated in a clinical sample of 43 female and 17 male IBS patients who completed the Eating Disorder Inventory (EDI). A diagnosis of IBS was generally unrelated to the Body Dissatisfaction, Perfectionism, and Ineffectiveness subscales of the EDI, but symptom severity was correlated with Perfectionism and Ineffectiveness. Severe bouts of vomiting were significantly associated with desires for lower body weight and reported binge-purge behaviors and cognitions measured by the Bulimia subscale of the EDI. Results suggest the need for a more comprehensive understanding of both types of illness as well as a possible framework for future empirical work. PMID- 9753390 TI - A conformational study in solution of pro-somatostatin fragments by NMR and computational methods. AB - The results of a conformational study by nuclear magnetic spectroscopy and computational methods on a series of point-mutated synthetic peptides, containing 14 amino acid residues and mimicking the region containing the Arg-Lys dibasic cleavage site of pro-somatostatin, have confirmed the possible role of a well defined secondary structure in the recognition phenomenon by processing enzymes. The importance of the residues located near the Arg-Lys dibasic site in the C terminal region of the pro-hormone for the cleavage of the precursor into somatostatin-14 has been confirmed. The present structural analysis indicates the occurrence of two beta-turns in the 4-7 and 11-14 regions, flanking the cleavage site, for all the peptides recognized as substrates by the processing enzyme. Interestingly, in the point-mutated analogue not processed by the enzyme and containing the replacement of proline by alanine in position 5 the first -turn is displaced by one residue and involves the Ala5-Arg8 segment. This observation may explain the lack of recognition by the maturation enzyme. PMID- 9753391 TI - Localisation of a protein core-specific epitope from gastrointestinal mucin (MUC2). The effect of epitope immobilisation on antibody recognition. AB - Human intestinal mucins are high molecular weight glycoproteins which protect and lubricate the epithelium of the gastrointestinal tract. In cases of malignant disease, mucins are abnormally expressed, overproduced or underglycosylated. This feature may enable the mucins to serve as tumour markers. The MUC2 mucin largely consists of a variable number of tandem repeats of a 23 amino acid sequence, 1PTTTPITTTTTVTPTPTPTGTQT23. In this study we have localised the minimal and the optimal epitope within this region by the previously developed protein core specific 996 monoclonal antibody using synthetic peptides. Several overlapping and truncated peptides related to the tandem repeat unit have been prepared by solid-phase methodology. Other mucin peptides were synthesised on the tips of polyethylene pins, and these remained C-terminally attached to the pins for comparative investigations. The interaction of the 996 monoclonal antibody with the synthetic peptides was studied either in solution by competition RIA or on immobilised peptides by indirect ELISA experiments. These experiments show that the minimal epitope recognised by the 996 antibody is the Ac-19TGTQ22 (IC50=3100 microM in solution). For the optimal 996 antibody binding in solution the 16PTPTGTQ22 heptapeptide (IC50 = 3 microM) is required. PMID- 9753392 TI - Large-scale production of peptides using the solid phase continuous flow method. Part 2: preparative synthesis of a 26-Mer peptide thrombin inhibitor. AB - A preparative method for the preparation of large peptides is described. An advantageous theoretical weight of peptide/weight of starting resin ratio (tPw/Rw) of about 0.3 was successfully experimented. The esterification of the first amino acid was realized with a racemization of less than 1%. The study of the coupling conditions led to the use of a diluted acylating mixture that allowed a 56% consumption of the amino acid derivatives (percentage use of amino acids) introduced in the synthesis. The cost analysis of the synthesis showed that the recovery of the amino acid derivatives was not worthwhile. PMID- 9753393 TI - Solid phase synthesis of cyclic peptides: model studies involving i-(i+4) side chain-to-side chain cyclisation. AB - Conditions for the synthesis of i-(i + 4) side chain-to-side chain head-to-tail Lys-->Glu and Glu-->Lys linked cyclic peptides related to hypoglycaemic analogues of human growth hormone hGH [6-13] have been examined. The success of the cyclisation reaction with the corresponding resin-bound, partially protected linear peptides was found to be both reagent as well as sequence dependent, with competing inter-chain oligomerisation predominating in some cases. The results also indicated that protection with the bulky Fmoc group of the amino acid residues immediately adjacent to the side chain-deprotected Lys and Glu residues, which participate in the cyclisation reaction, enhanced the rate of lactam formation. PMID- 9753394 TI - Influence of the secondary structure on the pore forming properties of synthetic alamethicin analogs: NMR and molecular modelling studies. AB - Synthetic alamethicin analogs, in which all Aib residues had been replaced by Leu (L2) then proline 14 replaced by an alanine (L5), were studied in SDS micelles using circular dichroism and NMR spectroscopy. Nuclear Overhauser effects were used as constraints for molecular modelling. The structures determined for both peptides in SDS micelles were compared with those previously obtained in methanol in order to establish a secondary structure/ionophore activity relationship. Our results indicated that a shortening of peptide helices could be responsible for the observed decrease in ion channel lifetimes. However, the length of helices may not by itself explain the drastic destabilization of channels when Pro14 of alamethicin is replaced by Ala in L5. Indeed analysis of the helical wheel of L5 reveals heterogeneity in the amphipathicity depending on the medium. Thus, loss of amphipathicity seems to underly the observed destabilization of channels. PMID- 9753395 TI - Synthesis and structural characterization of human-identical lung surfactant SP-C protein. AB - An efficient synthesis for human-identical lung surfactant protein SP-C is described with a semi-automated solid phase synthesizer using Fmoc chemistry. Double coupling and acetic anhydride capping procedures were employed for synthetic cycles within the highly hydrophobic C-terminal domain of SP-C. Isolation of the protein was performed by mild cleavage and deprotection conditions and subsequent HPLC purification yielding a highly homogeneous protein as established by sequence determination, electrospray, plasma desorption and MALDI mass spectrometry. A general method has been employed for the preparation of Cys-palmitoylated protein by using temporary Cys(tButhio) protection, in situ deprotection with beta-mercaptoethanol and selective palmitoylation of resin bound SP-C. The mild synthesis and isolation conditions provide SP-C with a high alpha-helical content, comparable to that of the natural SP-C, as assessed by CD spectra. Furthermore, first biophysical data indicate a surfactant activity comparable to that of the natural protein. PMID- 9753396 TI - Trans-cis amide bond isomerization in fulleroprolines. AB - The 1H NMR study of fulleroproline derivative Ac-Fpr-OtBu and its Pro analogue Ac L-Pro-OtBu over a range of temperatures in toluene-d8 solution has enabled the comparison of their equilibrium and activation parameters for the trans/cis interconversion around the amide partial double bond. PMID- 9753397 TI - Thalamic tumors in children. PMID- 9753398 TI - Meningiomas of the lateral ventricles of the brain in children. AB - Meningiomas of the lateral ventricles of the brain are rare tumours, accounting for approximately 0.5-5% of all intracranial meningiomas. Their natural history and symptomatology and the possibilities of early diagnosis are presented. The intraventricular location of the slow-growing benign mass provides a compensatory mechanism in the form of reserve space, which contributes to the delay in clinical demonstration of symptoms and signs. This makes the choice of diagnostic procedure an essential problem. CT and MRI are useful in detecting these masses, and magnetic resonance angiography (MRA) has also proved to be of great value in demonstrating the vascular supply of the tumour. This paper deals with two cases. In case 1 CT, MRI and MRA and in case 2 CT examination proved to be very useful. The tumours were removed by a transcortical approach in the posterior area. PMID- 9753399 TI - Administration of methylprednisolone acetate into the subdural cavity in an infant with subdural fluid collection. PAF is a good indicator of the efficacy of the treatment. AB - We report on the successful treatment of a case of infantile subdural fluid collection after cardiac surgery by administration of methylprednisolone acetate into the subdural cavity. The protein content in subdural fluid did not change even after administration of this steroid and was not a good indicator of the efficacy of treatment; however, the content of platelet-activating factor (PAF), an inflammatory mediator, in the fluid removed from the subdural cavity decreased rapidly after administration of the steroid. Subdural fluid collection subsequently decreased and mental retardation was improved. Our findings suggest that PAF content is a good indicator of the severity of infantile subdural fluid collection. PMID- 9753400 TI - Prognosis of ependymoma. AB - Ependymoma is a rare tumor entity. It affects children and adults and has four preferential locations: supra- and infra-tentorial, spinal, and conus-cauda filum. For statistical reasons, therefore, it is difficult to identify prognostic factors in series from single institutions because of the limited number of cases. The problem is complicated by the existence of some variants and by the difficulty in recognizing the anaplastic variant, because the common criteria used for recognizing malignancy in gliomas are not completely reliable in ependymomas. Apart from age and location, many parameters have been considered for prognostic purposes, including extension of surgical removal and radiotherapy. Among histological factors, the number of mitoses, labelling indices of proliferation markers, and cell density turned out to be good parameters for prognostic purposes. An overview is given of the contributions in the literature, including our own, on this problem. PMID- 9753401 TI - Long-term socioeconomic outcome following surgical intervention in the treatment of refractory epilepsy in childhood and adolescence. AB - Surgical treatment of refractory epilepsy in childhood and adolescence has been shown to be effective in reducing the seizure frequency. This paper examines the question: "Does this result in a better socioeconomic outcome in later years?" Patients who underwent a surgical procedure for the treatment of their medically refractory epilepsy at our hospital, had more than 2-years' follow-up, and were less than 18 years old at time of survey were included. From a retrospective chart review, age at onset and at surgery, duration of seizures prior to surgery, years of follow-up, type of surgery, and neurological status were obtained. From a telephone survey, seizure frequency after surgery, marital, financial and driving status, level of education, and employment status were ascertained. Sixty four patients in our epileptic surgical series meet entry criteria. Significantly higher levels of education, employment status and independence were found in patients with a class I Engel outcome compared to other Engel outcomes. PMID- 9753402 TI - Regional cerebral blood flow in pediatric moyamoya disease: age-dependent decline in specific regions. AB - Thirteen pediatric patients (ages 4-13 years) who underwent surgical treatment were examined regarding their rCBF in the preoperative periods. The postoperative rCBF was measured 39 times in these 13 patients. Thirteen healthy normal subjects (ages from 6 to 21 years) were also examined. The rCBF in the operculum and in the frontal, parietal, and occipital lobes was measured with 133Xe inhalation method and single photon emission computed tomography. In the parietal and occipital lobes, the preoperative rCBF had a negative and significant correlation with their ages, but not in the operculum or frontal lobe. However, subsequent to the surgical treatment, the rCBF increased significantly in the patients 5 years old or less, and then post-operative rCBF values had significant negative correlations with age in each region. PMID- 9753403 TI - Two different surgical techniques for reduction cranioplasty. AB - Reduction cranioplasty is required in selected patients when macrocephaly interferes with head control, seating, locomotion, and social acceptance. Two different surgical techniques for reduction cranioplasty in two cases of older hydrocephalic patients are described. Emphasis is placed on the basic stages of the procedure. PMID- 9753404 TI - Propionibacterium [correction of Proprionibacterium] acnes infections of cerebrospinal fluid shunts. AB - Ventricular cerebrospinal fluid shunt infections with Propionibacterium acnes are generally low-grade, indolent infections. Typical presentations include gradual shunt malfunction, nausea, headache, malaise, and infrequently, fever. In all, 489 shunt procedures performed between January 1992 and December 1995, and in 15 of these cases P. acnes was subsequently cultured from reservoir taps or an intraoperative culture which was obtained when the existing shunt components were revised. Six of these, representing 14.6% of shunt infections, were considered to be true P. acnes shunt infections, as they were associated with either CSF leukocytosis or the identification of gram-positive rods by gram stain. The others were considered to be probable contaminants. Detailed analysis of all 15 of these cases revealed that no patient had positive CSF cultures after removal of the infected shunt and the initiation of antibiotics. Given the benign characteristics of P. acnes shunt infections, the broad sensitivity to antibiotics, and the rapid sterilization of the cerebrospinal fluid, it may be possible to treat such cases with short-term perioperative antibiotics and replacement of the shunt components, in place of prolonged external ventricular drainage and antibiotics. This would have eliminated 8 operative procedures and reduced the estimated length of stay by 77 patient-days in these 15 children. PMID- 9753405 TI - Principles and concepts of brain death and organ donation: the Jewish perspective. AB - The harvesting of organs for transplantation is dependent on a stringent definition of brain death. Different societies have had to struggle with their cultural heritage, adapting their traditional attitudes to conform to the advances in medical science and the needs of the sick. In this article, the development of the concept of brain death as it applies to organ transplantation in Judaism is outlined. The ability of traditional Jewish values to address themselves to the challenges of modern medicine can serve as a basis for cultural cross-fertilization and comparison in modern societies. PMID- 9753406 TI - Diffuse bilateral thalamic astrocytomas as examined serially by MRI. AB - We report the case of a 13-year-old girl with diffuse bilateral thalamic astrocytomas. Incoordination was observed at the onset. Cranial computed tomography (CT) showed enlarged thalami, and magnetic resonance imaging (MRI) revealed these lesions to be symmetrically enlarged with high intensity on the T2 weighted image. Owing to these atypical findings in the neuroimaging studies, we had difficulty in making the correct diagnosis of a brain tumor. After the diagnosis of diffuse bilateral thalamic astrocytomas was obtained, we performed hyperfractionated radiotherapy followed by chemotherapy. Radiation therapy was effective for a while, but the girl's condition deteriorated again and she died 8 months after admission. Although diffuse bilateral thalamic astrocytomas are difficult to diagnose because they do not resemble most other neoplasms on neuroimaging studies, pediatricians should keep this entity in mind in order to arrive at a precise and prompt diagnosis. PMID- 9753407 TI - Melanotic progonoma of the brain: a case report and review. AB - So far, only 25 melanotic progonomas have been found in the central nervous system (male/female ratio 3.5), mostly located in the cerebellum. The average age is 8 years (range 3.5 months to 69 years) with 85% becoming clinically apparent in the first decade of life; 73.7% of the patients reported succumbed to their disease at a mean age of 2.8 years, with a postoperative survival time of just 9 months. Systemic metastases were reported in 9 cases and had mostly spread via cerebrospinal fluid. In contrast to peripheral melanotic progonomas usually found in the maxilla, cerebral progonomas have a much worse outcome and have to be regarded as malignant. We present the case of a 1-year-old boy suffering from a melanotic progonoma of the pineal gland, who died at the age of 22 months with extensive spinal and abdominal metastases 10 months after partial removal of the tumor. PMID- 9753408 TI - Herniation of cerebellar tonsils following supratentorial shunt placement. AB - Acquired Chiari 1 following ventriculoperitoneal shunting is an extremely unusual event. We report the case of an 8-year-old boy who presented with clinical and radiological signs of cerebellar tonsil herniation shortly after the placement of a cystoperitoneal shunt. Quantitative analysis of posterior fossa volumes (PFV) revealed that the patient had a smaller posterior fossa than age-matched normal controls. This abnormality, expressed as a decreased ratio between the posterior fossa and the supratentorial cavities (PFR), had already been present when the preoperative MRI was done. Our results suggest that preexisting structural abnormalities in the posterior fossa may constitute an important factor in the development tonsillar herniation following supratentorial shunts. PMID- 9753409 TI - A split cord malformation. AB - This report describes a case of split cord malformation without a septum. A 2 year-old boy presented with a 3-month history of neurogenic bladder. MRI did not show any apparent abnormality around the conus medullaris. However, CT myelography clearly demonstrated the presence of a split filum terminale. The patient underwent laminectomy of L1-5 laminas and untethering of the split filum terminale. CT myelography was superior to MRI in diagnosing split cord malformation in this case. As MRI is currently regarded as the superior imaging modality, this reported case may have been missed because the pathology was not apparent on MRI. PMID- 9753410 TI - Hepatic and extrahepatic clearance of circulating human lactoferrin: an experimental study in rat. AB - Lactoferrin, unlabelled or 125I-labelled by 2 different methods, was given intravenously to rats. Blood, tissue and liver cell radioactivity was measured. Both of the radiolabelled preparations were eliminated by the liver, and some deposited extrahepatically. One preparation formed large aggregates--here 90% of the hepatic uptake occurred in the Kupffer cells. The other preparation, consisting mostly of protein monomers but also dimers/oligomers/microaggregates, was taken up by hepatocytes (63% of total liver uptake), liver endothelial cells (22%) and Kupffer cells (15%). On a per cell volume basis, lactoferrin uptake was much more efficient by nonparenchymal cells compared to hepatocytes, which explains why immunomorphological staining only revealed lactoferrin in the nonparenchymal liver cells. The study demonstrates that radio-iodination of lactoferrin can affect its properties and handling, which may be important regarding contradictory reports on hepatic lactoferrin uptake. We conclude that both hepatocytes and nonparenchymal liver cells are involved in the blood clearance of lactoferrin, probably to a great extent owing to nonspecific mechanisms. Extrahepatic deposition and exposure (for instance on vessel walls/glomeruli) suggests that lactoferrin can be available to circulating anti lactoferrin autoantibodies in autoimmune disease. PMID- 9753411 TI - Red cell folate in elderly patients with myelodysplastic syndrome. AB - During a 7-month period a prospective study of 71 anaemic patients (29 males and 42 females) over the age of 50 was undertaken in order to identify patients with myelodysplastic syndrome (MDS). The mean values of mean corpuscular volume (MCV), serum ferritin, folate, vitamin B12 and red cell folate (RCF) of patients grouped according to the diagnosis were compared to those observed in age-matched blood donors. Forty-four of the 71 elderly patients showed macrocytic anaemia: 21 of them had gastric disease and the remaining 23 MDS. Two further patients with MDS showed microcytic anaemia. The 25 patients diagnosed with MDS were subclassified according to the FAB nomenclature: 9 had a refractory anaemia with excess of blasts and 16 refractory anaemia. The mean values of MCV, serum folate, ferritin, vitamin B12 and RCF were statistically different between patients with macrocytic anaemia due to gastric disease and patients with MDS. Among patients with MDS, the RCF level pathologically high was inversely correlated to the haemoglobin level (r=-0.39; p<0.05). Thus the RCF and serum folate may represent useful parameters for the diagnosis of MDS in elderly anaemic patients. PMID- 9753412 TI - Low rate of somatic hypermutations characterize progressive B-cell lymphomas. AB - Immunoglobulin heavy (IgH) chain gene rearrangements were characterized in 40 samples from 15 patients with B-cell lymphomas at different time points during tumour progression. Using polymerase chain reaction (PCR) amplification and single strand conformation polymorphism (SSCP) analysis of variable heavy (VH) chain gene segments, we found that 6 cases displayed alterations in their IgH chain rearrangements at relapse. These alterations were mainly observed in follicular or transformed lymphomas, but no association to clinical features was found. Nucleotide sequence analysis revealed a low frequency of mutations in 3 cases, whereas 1 case displayed an extensive mutation rate in a compartment with transformed morphology at relapse. The mutations observed most probably resulted from somatic hypermutations. Further, the mutations were scattered randomly over the VH gene segment and no significant bias favouring amino acid substitutions was observed in 3 cases, suggesting that the tumour cells had not been subjected to antigen-driven selection. In 1 case, however, the mutation pattern indicated that the tumour cells had been affected by an antigen selection process. In the 2 remaining cases, the original V(H)DJ(H) rearrangement could no longer be detected by VH gene family specific PCR at relapse, but using primers specific for the framework region 2 or 3 altered rearrangements were demonstrated, implying that mutations had been introduced in framework region 1. However, the majority of the tumour cell clones analysed were relatively stable during tumour progression, which make them eligible for analysis of minimal residual disease using the VH gene regions as molecular markers. PMID- 9753413 TI - BEAM+autologous stem cell transplantation in malignant lymphoma: 100 consecutive transplants in a single centre. Efficacy, toxicity and engraftment in relation to stem-cell source and previous treatment. AB - One hundred consecutive patients with malignant lymphoma treated with high-dose chemotherapy and autologous stem cell transplantation, followed at least 1 yr post-transplant, are reported, 68 with non-Hodgkin's lymphoma and 32 with Hodgkin's disease. At transplant, 23 patients were in first remission, 69 in later chemosensitive disease and 8 were chemotherapy resistant. Based on previous treatment and stem-cell source, the patients were subdivided into 3 cohorts: BMT1: bone-marrow harvest and transplant after > or =3 treatment regimens (38 patients); BMT2: bone marrow harvest and transplant after less than 3 treatment regimens (24 patients); PBSCT: peripheral-blood stem cell transplant (38 patients, 5 of these with CD34+ cell selected PBSC). The 4-yr survival and progression-free survival of all patients was 45 and 40%, respectively. Forty-one patients have died, 27 of lymphoma, evenly distributed in the cohorts. Fourteen treatment-related deaths occurred, 13 of these in the BMT1 cohort, significantly more than in the other cohorts (p=0.001). In univariate survival analysis cohort, age, disease status at transplant and number of previous treatment regimens were significant. In multivariate survival analysis cohort, age and sex were independently significant, women having a shorter survival. The patients transplanted with unselected PBSC had significantly shorter duration of pancytopenia and hospital stay than the otherwise comparable BMT2 patients, but their progression-free survival was identical. We confirm that high-dose therapy with autologous stem cell transplant from blood or bone marrow in not-too-heavily pretreated patients is a safe procedure but will cure only half the patients. PMID- 9753414 TI - Genetic polymorphism H131R of Fcgamma receptor type IIA (FcgammaRIIA) in a healthy Finnish population and in patients with or without platelet-associated IgG. AB - Patients (n=113) with histories of thrombocytopenia and with different profiles for platelet-associated IgG (PA-IgG) were subdivided according to the genetic polymorphism H131R in the Fcgamma receptor type IIA (FcgammaRIIA). PA-IgG was measured by the direct platelet immunofluorescence test (PIFT), and GP IIbIIIa and/or GP Ib-specific PA-IgG was investigated by a modified version of the direct monoclonal antibody-specific immobilization of platelet antigens (MAIPA) assay. As a control, the distribution of FcgammaRIIA polymorphism H131R was determined among 93 healthy Finnish blood donors. The frequencies for H131 and R131 were 0.56 and 0.44 (CI: 0.37-0.51), respectively, which did not differ significantly from those in other Caucasian populations. The distribution of the genotypes HH131, HR131 and RR131 in the patients and controls did not differ significantly. In the HH131 group, the PA-IgG was higher than in the RR131 group (p=0.082). Female patients with the genotype RR131 seemed to be younger than those with HH131 (p=0.065). Among the female patients, a significantly greater number were under 40 yr old in the RR131 group than in the HH131 group (p=0.0060). Within the RR131 group, the female patients were far younger than the male patients (median 29 vs. 61 yr; p=0.0021). The results point to the heterogeneity of immune thrombocytopenia, which may partly explain the poor predictive value of PA-IgG studies. PMID- 9753415 TI - Signaling through interleukin-6 receptor supports blast cell proliferation in acute myeloblastic leukemia. AB - The role of the interleukin-6/interleukin-6 receptor (IL-6/IL-6R) system in regulating blast cell growth in 8 acute myeloblastic leukemia (AML) patient derived cell lines was investigated. As they all expressed IL-6R and as none of them responded to exogenous IL-6 under conventional serum-supplemented culture conditions, we investigated whether signaling through IL-6R plays any role in maintaining their spontaneous colony growth. This was done by treating the cells with monoclonal antibodies made against the ligand-specific IL-6R alpha-chain or the signal transducer gp130. In serum-supplemented cultures inhibition of gp130 function did not affect the cell line growth, whereas anti-IL-6R alpha-chain antibody reduced colony growth. While some of the cell lines also showed similar growth characteristics in a serum-free environment, some others changed their growth pattern and stopped responding to anti-IL-6R alpha-chain treatment. At the same time, these cell lines also began to respond to exogenously added IL-6 and, interestingly, were stimulated by anti-gp130 antibody. Hence, in some of the blast cells, clonogenic cell growth seemed to be also negatively controlled by an endogenously produced growth-depressing cytokine or cytokines that utilize gp130. All the cell lines, whether cultured in the presence or absence of serum expressed IL-6 both at mRNA and protein level. The current results indicate that AML cells can use IL-6 as a growth stimulating factor, supplied either paracrinely or autocrinely. This could implicate the use of anti-IL-6R alpha chain antagonists in AML treatment, not IL-6. PMID- 9753417 TI - Avascular osteonecrosis in patients treated for Hodgkin's disease. AB - The aim of this study is to assess the risk of avascular osteonecrosis (AVN) of the femoral head in patients treated for Hodgkin's disease (HD), in relation to the type of treatment they have received. For this purpose, a cohort of 1391 patients treated for HD at University of Rome between 1972 and 1996 was divided into 2 groups according to their initial treatment. The first group contained 784 patients treated, at the onset of HD, either with chemotherapy (CT) containing steroids, combined in some cases with subdiaphragmatic radiotherapy (RT), or with subdiaphragmatic RT combined with CT without steroids. The second group was made up of 607 patients who had received, initially, supradiaphragmatic RT alone or supradiaphragmatic RT combined with CT without steroids. For the purpose of this study, only the 784 patients belonging to the first group were observed for the appearance of AVN, which occurred in 9 cases. The period of time which elapsed between the end of treatment and the radiological evidence of AVN ranged from 23 to 97 months, with an average of 35 months. Because the number of cases of AVN was so small, the pathogenesis of this complication could not be identified. PMID- 9753416 TI - First line Fludarabine treatment of symptomatic chronic lymphoproliferative diseases: clinical results and molecular analysis of minimal residual disease. AB - Fludarabine (25 mg/m2 for 5 d, every 4 wk, for 6 courses) was administered as first line therapy in 32 symptomatic chronic lymphoproliferative diseases. All CLL patients achieved at least partial response (5 CR, 2 nPR, 9 PR) but 44% of patients relapsed. In LG-NHLs response and relapse rate were similar. Haematological toxicity was low. VDJ rearrangement PCR analysis was performed on marrow samples at diagnosis and at the time of response evaluation. In the 3 patients who underwent high dose therapy with peripheral blood progenitor cell rescue analysis was also performed on apheresis samples and on marrow samples at the end of the procedure. Clonal VDJ rearrangement was always evident after Fludarabine therapy even in those patients who achieved histological and immunophenotypic complete remission, whereas it disappeared in 2 of 3 patients who underwent HDT. Our data confirm that Fludarabine monotherapy can reduce the neoplastic mass to a subclinical level and suggest the possibility that high dose therapy might produce true complete remission. PMID- 9753418 TI - Transferrin receptor-ferritin index: a useful parameter in differential diagnosis of iron deficiency and hyperplastic erythropoiesis. PMID- 9753419 TI - High-dose intravenous immune globulin and the response to splenectomy monitoring with platelet-associated IgG in patients with idiopathic thrombocytopenic purpura. PMID- 9753420 TI - Normal platelet numbers correlate with plasma viral load and CD4+ cell counts in HIV-1 infection. PMID- 9753421 TI - Drug-induced pneumonia associated with hemizygote glucose-6-phosphate dehydrogenase deficiency. PMID- 9753422 TI - Successful treatment with cyclosporine A of pure red-cell aplasia occurring 11 years after thymectomy. PMID- 9753423 TI - Meiotic telomeres: a matchmaker for homologous chromosomes. AB - Telomeres, with their special structures and special schemes of synthesis, are essential for protecting the ends of eukaryotic linear chromosomes during cell proliferation. In addition to this basic function, the meiosis-specific functions of telomeres have long been inferred from the cytological observations of characteristic chromosome configurations in meiotic prophase. Recent studies in the fission yeast Schizosaccharomyces pombe have provided deeper insights into the role of meiotic telomeres in the pairing of homologous chromosomes. Here I have summarized our current understanding of the meiotic behaviour of telomeres in S. pombe, and discuss the role of telomeres in meiosis. PMID- 9753424 TI - Individual LCR hypersensitive sites cooperate to generate an open chromatin domain spanning the human beta-globin locus. AB - BACKGROUND: The human beta-globin locus control region (LCR) is composed of five DNase I hypersensitive (HS) sites located 5' to the multiple genes it regulates. The LCR has been shown to comprise, among other essential properties, an activity that is required for generating a chromatin structure which renders the entire beta-globin gene locus accessible to exogenous nucleases. This nuclease-sensitive state is generally believed to be reflective of the chromatin environment that is permissive for transcriptional activation of the globin genes. RESULTS: Here we show, in mice bearing intact YAC transgenes that encompass the whole human beta globin locus, that the deletion of individual core LCR HS sites negatively affects the ability of the LCR to confer this open chromatin conformation throughout the locus, and when analysed in concert with the effect that these same mutations have on transcription, the data show that the chromatin opening activity is a necessary, but not sufficient, prerequisite for globin gene expression. The results also show that after deletion of individual hypersensitive sites, the mutated LCR is no longer able to provide an accessible chromatin environment that is independent from the site of YAC transgene integration. CONCLUSIONS: These experiments provide further evidence for the hypothesis that the HS sites must act cooperatively to fulfil the multiple functions that are attributable to the LCR. PMID- 9753425 TI - Grb10/GrbIR as an in vivo substrate of Tec tyrosine kinase. AB - BACKGROUND: Tec is a member of the recently emerging subfamily among nonreceptor protein-tyrosine kinases (PTKs). Although many members of this family have been shown to be involved in a wide range of cytokine-mediated signalling systems, the molecular mechanism by which they exert in vivo effects remains obscure. To gain insights into the downstream pathways of Tec, we here looked for Tec-interacting proteins (TIPs) by using the yeast two-hybrid screening. RESULTS: One of TIPs turned out to be Grb10/GrbIR, which carries one pleckstrin homology domain and one Src homology 2 domain. Grb10/GrbIR was known to bind receptor PTKs in a ligand-dependent fashion, but not to be phosphorylated on tyrosine residues. In a transient expression system in human kidney 293 cells, however, Grb10/GrbIR becomes profoundly tyrosine-phosphorylated by Tec, but not by Syk, Jak2 or insulin receptor. We also reveal that expression of Grb10/GrbIR suppresses the cytokine-driven and Tec-driven activation of the c-fos promoter. CONCLUSION: Our results indicate a novel role of Grb10/GrbIR as an effector molecule to a subset of nonreceptor PTKs. PMID- 9753426 TI - Transcription factor CP2 is essential for lens-specific expression of the chicken alphaA-crystallin gene. AB - BACKGROUND: Lens-specific transcriptional activation of the chicken alphaA crystallin gene is controlled by the distal and proximal enhancers, alphaCE1 and alphaCE2, respectively. Analysis using specific monoclonal antibodies against purified alphaCE1-binding factor alphaCEF1 revealed that alphaCEF1 is composed of two distinct subunits. RESULTS: We have demonstrated that one of the subunits of alphaCEF1 is encoded by chicken ubiquitous transcription factor CP2 (cCP2), which is homologous to mouse CP2, and human CP2/LBP-1/LSF-1. Electrophoretic mobility shift assays and cross-linking experiments showed that alphaCEF1 and bacterially expressed cCP2 form a tetramer. Overexpression of cCP2 activates transcription through alphaCE1, but a mutant cCP2 lacking the DNA-binding domain reduced the transcription to basal levels. Although cCP2 binds to the CP2 template from the mouse alpha-globin promoter, it fails to promote transcription through this template. Element substitution experiments between alphaCE1 and the CP2 template revealed that the lens-specific enhancer activity of alphaCE1 is due to the 6 bp sequence (-139/-134; lens-specific element (LSE)) adjacent to the 3' of the cCP2 binding site within alphaCE1. CONCLUSION: We have shown that the tetrameric transcription factor cCP2 is essential for lens-specific transcription of the chicken alphaA-crystallin gene, although it is ubiquitously expressed. We propose a model where cCP2 cooperates with a putative lens-specific factor which binds to LSE. PMID- 9753427 TI - Large scale isolation of osteoclast-specific genes by an improved method involving the preparation of a subtracted cDNA library. AB - BACKGROUND: Osteoclasts play crucial roles in bone resorption, which triggers bone remodeling. Molecular mechanisms underlying these osteoclast-specific biological functions remain elusive because only a limited number of osteoclast specific genes have been identified. To circumvent this, we isolated a large number of osteoclast-specific genes by preparing a subtracted cDNA library of high quality. RESULTS: We first constructed a plasmid expression vector (pAP3neo) that allowed an efficient subtraction. Then, we improved the standard protocols for preparation of the cDNA library and the subsequent subtraction procedure. Using our protocol, we prepared a rabbit osteoclast cDNA library of high complexity. Subsequently, we prepared an osteoclast-specific cDNA library of high complexity by subtracting it with biotin-labelled mRNA, derived from rabbit spleen through the biotin-avidin method. The resulting library included a high proportion of full-length cDNA inserts. Using DNA dot blot analysis, we found that the osteoclast-specific cDNA clones were highly enriched in this subtracted cDNA library, i.e. nearly 70% of the analysed clones were primarily detected in osteoclasts but not in spleen. Multiple-tissue Northern analysis also showed that many of these clones were expressed almost exclusively in osteoclasts. DNA sequencing of randomly selected clones showed that 424 cDNA species out of 1136 analysed were novel. DNA sequencing also showed that our subtracted cDNA library was almost equalized, suggesting that the library may contain almost all of the osteoclast-specific genes. CONCLUSION: From these data, we conclude that our subtraction protocols, and the subsequent procedure for the analysis of the isolated clones developed here, are useful for the comprehensive isolation and identification of transcriptionally up- or down-regulated genes in general. PMID- 9753428 TI - Post-transcriptional control of the level of mRNA by hepatitis B virus X gene in the transient expression system using human hepatic cells. AB - BACKGROUND: Hepatitis B virus (HBV) infection is closely related to the development of not only acute or chronic hepatitis, but also hepatocellular carcinoma. Among the HBV genes, the X gene has been implicated in the carcinogenicity of this virus as a major causative factor by its ability to activate viral and cellular genes in trans via protein-protein interaction with cellular factors without binding to DNA. RESULTS: To explore the possibility of other functions of the X gene, we examined the effect of X protein on the transient expression system of simian virus 40 (SV40) large T-antigen or chloramphenicol acetyltransferase (CAT) mRNA using SV40 promoter or EF-1alpha (human elongation factor 1alpha) promoter, by co-transfecting an X gene expression plasmid to human hepatic cell lines, HepG2 and Huh7. In contrast to the SV40 promoter-mediated expression, the level of both T-antigen and CAT mRNAs expressed from the EF-1alpha promoter was strikingly decreased by X protein in both hepatic cells. The nuclear run-on assay and the mRNA decay experiment using actinomycin D, indicated that the effect of X protein on the lowering of the level of chimeric mRNA was due to the degradation of mRNA, but not repression of transcriptional initiation. Moreover, this effect was dependent on the 22 bp sequence in the 5' untranslated region of mRNA derived from the EF-1alpha promoter. CONCLUSION: The present data suggest a new function of the X gene to post-transcriptionally control the stability of mRNA through the 5' untranslated region derived from the EF-1alpha promoter in human hepatic cells. PMID- 9753429 TI - Crystal structure of tyrosine hydroxylase with bound cofactor analogue and iron at 2.3 A resolution: self-hydroxylation of Phe300 and the pterin-binding site. AB - TyrOH is a non-heme iron enzyme which uses molecular oxygen to hydroxylate tyrosine to form L-dihydroxyphenylalanine (L-DOPA), and tetrahydrobiopterin to form 4a-hydroxybiopterin, in the rate-limiting step of the catecholamine biosynthetic pathway. The 2.3 A crystal structure of the catalytic and tetramerization domains of rat tyrosine hydroxylase (TyrOH) in the presence of the cofactor analogue 7,8-dihydrobiopterin and iron shows the mode of pterin binding and the proximity of its hydroxylated 4a carbon to the required iron. The pterin binds on one face of the large active-site cleft, forming an aromatic pi stacking interaction with Phe300. This phenylalanine residue of TyrOH is found to be hydroxylated in the meta position, most likely through an autocatalytic process, and to consequently form a hydrogen bond to the main-chain carbonyl of Gln310 which anchors Phe300 in the active site. The bound pterin forms hydrogen bonds from N-8 to the main-chain carbonyl of Leu295, from O-4 to Tyr371 and Glu376, from the C-1' OH to the main-chain amides of Leu294 and Leu295, and from the C-2' hydroxyl to an iron-coordinating water. The part of the pterin closest to the iron is the O-4 carbonyl oxygen at a distance of 3.6 A. The iron is 5.6 A from the pterin 4a carbon which is hydroxylated in the enzymatic reaction. No structural changes are observed between the pterin bound and the nonliganded enzyme. On the basis of these structures, molecular oxygen could bind in a bridging position optimally between the pterin C-4a and iron atom prior to substrate hydroxylation. This structure represents the first report of close interactions between pterin and iron in an enzyme active site. PMID- 9753430 TI - The 0.78 A structure of a serine protease: Bacillus lentus subtilisin. AB - Ultrahigh-resolution X-ray diffraction data from cryo-cooled, B. lentus subtilisin crystals has been collected to a resolution of 0.78 A. The refined model coordinates have a rms deviation of 0.22 A relative to the same structure determined at room temperature and 2.0 A resolution. Several regions of main chain and side-chain disorder have been identified for 21 out of 269 residues in one polypeptide chain. Hydrogen atoms appear as significant peaks in the Fo - Fc difference electron density map, and carbon, nitrogen, and oxygen atoms can be differentiated. The estimated standard deviation (ESD) for all main-chain non hydrogen bond lengths is 0.009 A and 0.5 degrees for bond angles based on an unrestrained full-matrix least-squares refinement. Hydrogen bonds are resolved in the serine protease catalytic triad (Ser-His-Asp). Electron density is observed for an unusual, short hydrogen bond between aspartic acid and histidine in the catalytic triad. The hydrogen atom, identified by NMR in numerous serine proteases, appears to be shared by the heteroatoms in the bond. This represents the first reported correlation between detailed chemical features identified by NMR and those in a cryo-cooled crystallographic structure determination at ultrahigh resolution. The short hydrogen bond, designated "catalytic hydrogen bond", occurs as part of an elaborate hydrogen bond network, involving Asp of the catalytic triad. While unusual, these features appear to have conserved analogues in other serine protease families although specific details differ from family to family. PMID- 9753431 TI - Structure determination of the Ras-binding domain of the Ral-specific guanine nucleotide exchange factor Rlf. AB - Ral-specific guanine nucleotide exchange factors RalGDS, Rgl, and Rlf have been suggested to function as intermediates between Ras and Ral pathways by being able to bind Ras proteins through their C-terminal Ras-binding domains (RBD). The RBDs of RalGDS and of the Ser/Thr kinase c-Raf-1 have been shown to have the same tertiary structure. In contrast to the RBDs of Raf and RalGDS, which bind either Ras or Rap with high affinity, Rlf-RBD has a similar affinity for both GTP binding proteins. To be able to compare these RBDs on a structural level, we have solved the three-dimensional structure of Rlf-RBD by NMR spectroscopy. The overall tertiary structure of Rlf-RBD shows the betabetaalphabetabetaalphabeta fold of the ubiquitin superfamily and is very similar to that of RalGDS-RBD. The binding interface of Rlf-RBD to Ras was mapped using chemical shift analysis and indicated a binding mode similar to that in the case of Rap.Raf-RBD. However, comparison of the putatively interacting regions revealed structural differences which are proposed to be responsible for the different substrate affinities of Rlf-, RalGDS-, and Raf-RBD. PMID- 9753432 TI - Location of the phosphate binding site within Clostridium symbiosum pyruvate phosphate dikinase. AB - Pyruvate phosphate dikinase (PPDK) catalyzes the interconversion of ATP, Pi, and pyruvate with AMP, PPi, and PEP in three partial reactions: (1) E + ATP --> E.ATP --> E-PP.AMP, (2) E-PP.AMP + Pi --> E-PP.AMP.Pi --> E-P.AMP.PPi, and (3) E-P + pyruvate --> E-P.pyruvate --> E.PEP. The Clostridium symbiosum PPDK structure consists of N-terminal, central, and C-terminal domains. The N-terminal and central domains catalyze partial reactions 1 and 2 whereas the C-terminal and central domains catalyze partial reaction 3. The goal of the present work is to determine where on the N-terminal domain catalysis of partial reactions 1 and 2 occurs and, in particular, where the Pi binding site is located. Computer modeling studies implicated Arg337 as a key residue for Pi binding. This role was tested by site-directed mutagenesis. The R337A PPDK was shown to be impaired in catalysis of the forward (kcat 300-fold lower) and reverse (kcat 30-fold lower) full reactions. Time courses for the single turnover reactions were measured to show that catalysis of partial reaction 1 is 5-fold slower in the mutant, catalysis of the second partial reaction is 140-fold slower in the mutant, and catalysis of the third partial reaction is unaffected. With the exception of the mutation site, the crystal structure of the R337A PPDK closely resembles the structure of the wild-type protein. Thus, the altered kinetic properties observed for this mutant are attributed solely to the elimination of the interaction between substrate and the guanidinium group of the Arg337 side chain. On the basis of these findings we propose that the Pi binding site is located within the crevice of the PPDK N-terminal domain, at a site that is flanked by the ATP beta P and the Mg2+ cofactor. PMID- 9753433 TI - Thermophilic xylanase from Thermomyces lanuginosus: high-resolution X-ray structure and modeling studies. AB - The crystal structure of the thermostable xylanase from Thermomyces lanuginosus was determined by single-crystal X-ray diffraction. The protein crystallizes in space group P21, a = 40.96(4) A, b = 52. 57(5) A, c = 50.47 (5) A, beta = 100.43(5) degrees, Z = 2. Diffraction data were collected at room temperature for a resolution range of 25-1.55 A, and the structure was solved by molecular replacement with the coordinates of xylanase II from Trichoderma reesei as a search model and refined to a crystallographic R-factor of 0.155 for all observed reflections. The enzyme belongs to the family 11 of glycosyl hydrolases [Henrissat, B., and Bairoch, A. (1993) Biochem. J. 293, 781-788]. pKa calculations were performed to assess the protonation state of residues relevant for catalysis and enzyme stability, and a heptaxylan was fitted into the active site groove by homology modeling, using the published crystal structure of a complex between the Bacillus circulans xylanase and a xylotetraose. Molecular dynamics indicated the central three sugar rings to be tightly bound, whereas the peripheral ones can assume different orientations and conformations, suggesting that the enzyme might also accept xylan chains which are branched at these positions. The reasons for the thermostability of the T. lanuginosus xylanase were analyzed by comparing its crystal structure with known structures of mesophilic family 11 xylanases. It appears that the thermostability is due to the presence of an extra disulfide bridge, as well as to an increase in the density of charged residues throughout the protein. PMID- 9753434 TI - NMR structure and dynamics of an RNA motif common to the spliceosome branch-point helix and the RNA-binding site for phage GA coat protein. AB - The RNA molecules that make up the spliceosome branch-point helix and the binding site for phage GA coat protein share a secondary structure motif in which two consecutive adenine residues occupy the strand opposite a single uridine, creating the potential to form one of two different A.U base pairs while leaving the other adenine unpaired or bulged. During the splicing of introns out of pre mRNA, the 2'-OH of the bulged adenine participates in the transesterification reaction at the 5'-exon and forms the branch-point residue of the lariat intermediate. Either adenine may act as the branch-point residue in mammals, but the 3'-proximal adenine does so preferentially. When bound to phage GA coat protein, the bulged adenine loops out of the helix and occupies a binding pocket on the surface of the protein, forming a nucleation complex for phage assembly. The coat protein can bind helices with bulged adenines at either position, but the 3'-proximal site binds with greater affinity. We have studied this RNA motif in a 21 nucleotide hairpin containing a GA coat protein-binding site whose four nucleotide loop has been replaced by a more stable loop from the related phage Ms2. Using heteronuclear NMR spectroscopy, we have determined the structure of this hairpin to an overall precision of 2.0 A. Both adenine bases stack into the helix, and while all available NOE and coupling constant data are consistent with both possible A.U base pairs, the base pair involving the 5'-proximal adenine appears to be the major conformation. The 3'-proximal bulged adenine protonates at unusually high pH, and to account for this, we propose a model in which the protonated adenine is stabilized by a hydrogen bond to the uridine O2 of the A.U base pair. The 2'-OH of the bulged adenine adopts a regular A-form helical geometry, suggesting that in order to participate in the splicing reaction, the conformation of the branch-point helix in the active spliceosome may change from the conformation described here. Thus, while the adenine site preferences of the spliceosome and of phage GA may be due to protein factors, the preferred adenine is predisposed in the free RNA to conformational rearrangement involved in formation of the active complexes. PMID- 9753435 TI - The active-site arginine of S-adenosylmethionine synthetase orients the reaction intermediate. AB - S-Adenosylmethionine (AdoMet) synthetase catalyzes the formation of AdoMet and tripolyphosphate (PPPi) from ATP and L-methionine and the subsequent hydrolysis of the PPPi to PPi and Pi before product release. Little is known about the roles of active-site residues involved in catalysis of the two sequential reactions that occur at opposite ends of the polyphosphate chain. Crystallographic studies of Escherichia coli AdoMet synthetase showed that arginine-244 is the only arginine near the polyphosphate-binding site. Arginine-244 is embedded as the seventh residue in the conserved sequence DxGxTxxKxI which is also found at the active site of inorganic pyrophosphatases, suggesting a potential pyrophosphate binding motif. Chemical modification of AdoMet synthetase by the arginine specific reagents phenylglyoxal or p-hydroxyphenylglyoxal inactivates the enzyme. ATP and PPPi protect the enzyme from inactivation, consistent with the presence of an important arginine residue in the vicinity of the polyphosphate-binding site. Site-specific mutagenesis has been used to change the conserved arginine 244 to either leucine (R244L) or histidine (R244H). In the overall reaction, the R244L mutant has the kcat reduced approximately 10(3)-fold, with a 7 to 10-fold increase in substrate Km values; the R244H mutant has an approximately 10(5)-fold decrease in kcat. In contrast, the kcat values for hydrolysis of added PPPi by the R244L and R244H mutants have been reduced by less than 2 orders of magnitude. In contrast to the wild-type enzyme in which 98% of the Pi formed originates as the gamma-phosphoryl group of ATP, in the R244L mutant the orientation of the PPPi intermediate equilibrates at the active site yielding equal amounts of Pi from the alpha- and gamma-phosphoryl groups of ATP. Thus, the active-site arginine has a profound role in the cleavage of PPPi from ATP during AdoMet formation and in maintaining the orientation of PPPi in the active site, while playing a lesser role in the subsequent PPPi hydrolytic reaction. PMID- 9753436 TI - Synthesis and characterization of more potent analogues of hirudin fragment 1-47 containing non-natural amino acids. AB - Hirudin is the most potent and specific inhibitor of thrombin, a key enzyme in the coagulation process existing in equilibrium between its procoagulant (fast) and anticoagulant (slow) form. In a previous study, we described the solid-phase synthesis of a Trp3 analogue of fragment 1-47 of hirudin HM2, which displayed approximately 5-fold higher thrombin inhibitory potency relative to that of the natural product [De Filippis, V., et al. (1995) Biochemistry 34, 9552-9564]. By combining automated and manual peptide synthesis, here we have produced in high yields seven analogues of fragment 1-47 containing natural and non-natural amino acids. In particular, we have replaced Val1 with tert-butylglycine (tBug), Ser2 with Arg, and Tyr3 with Phe, cyclohexylalanine (Cha), Trp, alpha-naphthylalanine (alphaNal), and beta-naphthylalanine (betaNal). The crude reduced peptides are able to fold almost quantitatively into the disulfide-cross-linked species, whose unique alignment (Cys6-Cys14, Cys16-Cys28, and Cys22-Cys37) has been shown to be identical to that of the natural fragment. The results of conformational characterization provide evidence that synthetic peptides retain the structural features of the natural species, whereas thrombin inhibition data indicate that the synthetic analogues are all more potent inhibitors of thrombin. In particular, Val --> tBug exchange leads to a 3-fold increase in binding, interpreted as arising from a favorable reduction of the entropy of binding, due to the presence of the more symmetric side chain of tBug relative to that of Val. The S2R analogue binds 24- and 125-fold more tightly than the natural fragment to the fast or slow form of thrombin. These results are explained by considering that Arg2 may favorably couple to Glu192, a key residue involved in the slow to fast transition, thus stabilizing the slow form. Replacement of Tyr3 with more hydrophobic residues having different side chain orientations and electronic structures improves binding by 2-40-fold, suggesting that nonpolar interactions and shape-dependent packing effects strongly influence binding at this position. Overall, these results provide new insights for elucidating the mechanism of hirudin-thrombin recognition at the molecular level and highlight new strategies for designing more potent and selective inhibitors of thrombin. PMID- 9753437 TI - Stepwise subunit interaction changes by mono- and bisphosphorylation of cardiac troponin I. AB - Four phosphorylation degrees of cardiac troponin I (cTnI) have been characterized, namely, a dephospho, a bisphospho, and two monophospho states. Here we describe for the first time a role of the monophosphorylated forms. We have investigated the interaction between the cardiac troponin subunits dependent on the phosphorylation state of cTnI by surface plasmon resonance (SPR) spectroscopy. The monophosphorylated forms were generated by mutating each of the two serine residues, located in human cTnI at positions 22 and 23, to alanine. Association and dissociation rate constants of binary (cTnI-cTnT and cTnI-cTnC) and ternary (cTnI/cTnC complex-cTnT) complexes were determined. Mono- and consecutive bisphosphorylation of cTnI gradually reduces the affinity to cTnC and cTnT by lowering the association rate constants; the dissociation rate constants remain unchanged. Phosphorylation also affects formation of the ternary complexes; however, in this instance, association rate constants are constant, and dissociation rate constants are enhanced. A model of cardiac troponin is presented describing an induction of distinct conformational changes by mono- and bisphosphorylation of cTnI. PMID- 9753438 TI - Unsaturated phospholipid acyl chains are required to constitute membrane binding sites for factor VIII. AB - Membranes containing phosphatidyl-L-serine (PS) and phosphatidylethanolamine (PE) greatly enhance the function of the enzymatic cofactor factor VIII. The mechanisms of enhanced function involve condensation of enzyme (factor IXa), activated cofactor (factor VIIIa), and substrate (factor X) at a common location and, most dramatically, activation of the assembled enzyme-cofactor complex. We asked whether unsaturated phospholipid (PL) acyl chains are necessary to constitute factor VIII binding sites or to activate the factor VIIIa-factor IXa complex. We found that membranes composed of saturated, dimyristoyl phospholipids had 20-fold fewer factor VIII binding sites and that these sites supported less than 5% normal activity of the factor VIIIa-factor IXa complex. Thrombin activated factor VIII bound to a similar number of membrane sites, and thrombin activation did not reduce the affinity for saturated membranes more than 2-fold so that the loss of functional activity is due to a requirement of the factor VIIIa-factor IXa complex for unsaturated acyl chains that exceeds the requirement for factor VIII binding alone. Replacement of dimyristoyl-PS, -PE, or -PC individually with the corresponding unsaturated phospholipid restored 75%, 60%, and 15%, respectively, of factor VIII binding sites but less than 10% of factor VIIIa-factor IXa activating activity. Lyso-PS did not support binding of factor VIII or function of the factor VIIIa-factor IXa complex even when PE and phosphatidylcholine contained unsaturated acyl chains. We conclude that the sn-2 acyl chain of PS and unsaturated phospholipid acyl chains are chemical requirements for constitution of fully functional factor VIII binding sites on phospholipid membranes. PMID- 9753439 TI - Thermodynamic analysis of the binding of the polyglutamate chain of 5 formyltetrahydropteroylpolyglutamates to serine hydroxymethyltransferase. AB - The thermodynamic parameters for the binding of 5-formyltetrahydrofolate (5-CHO H4PteGlun) and its polyglutamate forms to rabbit liver cytosolic serine hydroxymethyltransferase (SHMT) were determined by a combination of isothermal titration calorimetry and spectrophotometry. Binding of 5-CHO-H4PteGlun to SHMT exhibits both positive enthalpy and entropy, showing that binding is entropically driven. 5-CHO-H4PteGlu5 has a 300-fold increased affinity for SHMT compared to 5 CHO-H4PteGlu. This increase in affinity is due primarily to a decrease in the positive enthalpy with little change in entropy. A variety of anions inhibit the binding of 5-CHO-H4PteGlu5 with Ki values in the 10-20 mM range. Anions are ineffective inhibitors of 5-CHO-H4PteGlu binding to SHMT, showing that anions compete for the polyglutamate binding site. There was little difference in the Ki values for a series of dicarboxylic acids as inhibitors of 5-CHO-H4PteGlu5, suggesting that spacing of the negative charges may not be important in determining their effectiveness as inhibitors. Both the mono- and pentaglutamate derivatives of 5-CHO-H4PteGlun were cross-linked to SHMT by a carbodiimide reaction to Lys-450 which resides in a stretch of Lys, His, and Arg residues. PMID- 9753440 TI - Heme speciation in alkaline ferric FixL and possible tyrosine involvement in the signal transduction pathway for regulation of nitrogen fixation. AB - The pH-dependent behavior of the ferric forms of two soluble truncations of Rhizobium meliloti FixL, FixL (heme and kinase domains, functional), and FixLN (heme domain) are examined by UV-visible, resonance Raman, and electron paramagnetic resonance spectroscopy. Global analysis of UV-visible data indicates that the pKa for hydroxide binding is slightly higher in FixL than in FixLN. Spectroscopic data show that high-spin and low-spin hydroxide adducts of FixLN and FixL exist in a thermal spin-state equilibrium with a significant fraction of the heme in the high spin form at room temperature. FixLN and FixL differ from myoglobin and hemoglobin in that their hemes are not fully ligated by hydroxide ion under strongly alkaline conditions. In addition to the binding of hydroxide ion, both FixLN and FixL undergo additional alkaline transitions that involve the deprotonation of tyrosine residues. FixLN contains four tyrosine residues. One has a pKa of 9.6, which is indistinguishable from that for hydroxide binding to the heme. The other three tyrosines have pKas greater than 11. At pH 11, the alkaline species react with cyanide to yield the familiar low-spin cyanide adduct. Upon reduction of the heme iron, the alkaline forms of the FixL deletion derivatives are converted to their deoxy forms. Resonance Raman spectra reveal that the Fe-His stretching vibrations of deoxyFixLN and deoxyFixL are not measurably shifted from those of their neutral counterparts. Treatment of the alkaline deoxyFixLs with O2 yields the respective oxy forms. Spectroscopic evidence indicates that the loss of activity at elevated pH cannot be attributed solely to generation of a low-spin heme hydroxide. Involvement of one or more tyrosines in signal transmission between the heme and kinase domains of FixL is proposed. PMID- 9753441 TI - Hydrodynamic studies on the manganese-stabilizing protein of photosystem II. AB - The solution conformation of the manganese-stabilizing protein of photosystem II was examined by analytical ultracentrifugation. Sedimentation velocity and sedimentation equilibrium studies were performed. These experiments yielded values for of 2.26 S with a diffusion constant, D, of 7.7 x 10(-)7 cm2 s-1. This s value is significantly lower than the apparent s value of 2.6 S previously reported [Miyao, M., and Murata, N. (1989) Biochim. Biophys. Acta 977, 315-321]. The molecular mass of the protein, 26.531 kDa, was verified by MALDI mass spectrometry. The diffusion coefficient was also determined by dynamic light scattering. The z-weighted average of D was 6.8 x 10(-)7 cm2 s-1. This result was somewhat lower than that observed by analytical ultracentrifugation due to the presence of slowly diffusing components in the sample. A two-component exponential fit of the dynamic light scattering data, however, gave D = 7.52 x 10(-)7 cm2 s-1 for the major component of the sample, which is in excellent agreement with the value determined by analytical ultracentrifugation. The value of s, the apparent sedimentation coefficient, was found to depend on the concentration of the protein and varied about 4% per milligram of protein. This is a feature of proteins which are asymmetric in solution. This asymmetry was examined using both the v-bar and Teller methods. Both methods indicated a significant degree of asymmetry for the manganese-stabilizing protein. Our findings indicate that the prolate ellipsoid model for the manganese-stabilizing protein is elongated in solution, with approximate dimensions of about 12.6 nm x 3.0 nm, yielding an axial ratio of 4.2. PMID- 9753442 TI - Effects of carboxyl amino acid modification on the properties of the high affinity, manganese-binding site in photosystem II. AB - Our previous work using the "diphenylcarbazide (DPC)-inhibition assay" has identified four amino acid (two carboxyls and two histidyls) ligands to four Mn2+ bound with high affinity on Tris-washed photosystem II (PSII) membrane fragments [Preston and Seibert (1991) Biochemistry 30, 9615-9624, 9625-9633]. One of the ligands binds a photooxidizable Mn, specifically, and the others bind either nonphotooxidizable Mn2+, Zn2+, or Co2+ [Ghirardi et al. (1996) Biochemistry 35, 1820-1828]. The current paper shows the following: (a) the high-affinity photooxidizable Mn, which donates to the oxidized primary PSII donor (YZ*), is bound to a carboxyl residue with a KM = 1.5 microM or Kd = 0.94 microM in the absence of DPC, and a Ki = 1.3 microM in the presence of DPC (both steady-state and flash approaches were used); (b) if this carboxyl is chemically modified using 1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide hydrochloride (EDC), Mn2+ is photooxidized at a lower affinity (Kd = 25 microM) site that does not involve carboxyl ligands; (c) low-affinity Mn is photooxidized (possibly by YD*, the oxidized form of the alternative PSII donor) with a KM = 220 microM at a completely different site that also requires a carboxyl ligand; (d) photooxidation of high-affinity DPC by YZ* with a KM of 40-42 microM or Kd of 49 58 microM occurs at a site that does not require carboxyl residues; (e) photooxidation of low-affinity DPC with a KM = 1200 microM occurs at a site (possibly near YD) that is not affected by carboxyl modification with EDC. Due to the similarities between the binding of the high-affinity photooxidizable Mn to EDC-treated membranes and to PSII complexes from Asp170D1 mutants [Nixon and Diner (1992) Biochemistry 31, 942-948], we identify its carboxyl residue ligand as Asp170 on D1, one of the reaction-center proteins. The second carboxyl ligand identified using the DPC-inhibition assay binds Mn (but not a photooxidizable one), Zn, or Co ions. At least one of the two histidyl ligands (either His337 on D1 or another unidentified histidyl) that bind nonphotooxidizable, high-affinity Mn2+ also binds Zn2+ and Co2+. PMID- 9753443 TI - Use of a novel histidyl modifier to probe for residues on Tris-treated photosystem II membrane fragments that may bind functional manganese. AB - In this paper, we investigate the effects of histidyl amino acid modification on high-affinity Mn binding to photosystem II (PSII) using methods similar to those used in the preceding paper [Ghirardi et al. (1998) Biochemistry 37, 0000] for carboxyl amino acid modification. Given the rather low specificity of diethyl pyrocarbonate (DEPC) for histidine modification, we modified Tris-washed PSII membranes with a novel and more specific histidyl modifier, platinum(II) (2,2':6',2"-terpyridine) chloride (Pt-TP). Both the "diphenylcarbazide (DPC) inhibition assay" and single-turnover flash approaches were used. The concentration dependence of Pt-TP modification on steady-state measurements shows two types of interactions, each accounting for about half of the full effect. At concentrations <50 microM, Pt-TP modifies mostly histidyls and abolishes half of the observed Mn inhibition of DPC-mediated 2,6-dichlorophenolindophenol (DCIP) photoreduction (equivalent to two high-affinity, Mn-binding ligands). This effect can be blocked by addition of Mn2+ during Pt-TP modification. Double-modification experiments with DEPC and Pt-TP demonstrate that both modifiers affect the same observable histidyl residues in PSII. Above 50 microM, Pt-TP modifies mostly cysteines (or histidines in a more hydrophobic environment) and has an additional effect on the reducing side of PSII that (a) does not involve Mn binding and (b) results in the apparent abolishment of all four of the Mn-binding ligands detected by the DPC-inhibition assay. Single-flash experiments show that histidyl modification does not eliminate the binding of the high-affinity, photooxidizable Mn2+ to Asp170 on D1 (nor does it significantly affect high-affinity DPC photooxidation), but it does decrease the binding affinity (Kd) of that Mn from 0.6 to 1.5 microM, particularly at lower (<50 microM Pt-TP) concentrations. Double-modification experiments also demonstrate that the lower affinity, photooxidizable Mn-binding site, uncovered when the high-affinity site is modified with 1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide hydrochloride (EDC) [see Ghirardi et al. (1998)], is not associated with a histidyl ligand. Three nonphotooxidizable, high-affinity Mn2+ ions bind to a second carboxyl and two histidyl ligands, and these Mn are not photooxidized by a flash even when the ligand to the photooxidizable Mn is modified by EDC. Proteolytic enzyme studies indicate that the two histidyl ligands identified by the DPC-inhibition assay are probably His337 on D1 and His 339 on D2, but His 332 on D1 is not eliminated. PMID- 9753444 TI - The distal cavity structure of carbonyl horseradish peroxidase as probed by the resonance Raman spectra of His 42 Leu and Arg 38 Leu mutants. AB - CO ligation to horseradish peroxidase C (HRPC) was studied by means of site directed mutagenesis and resonance Raman spectroscopy. The CO complexes of HRPC His 42 --> Leu and Arg 38 --> Leu mutants were characterized at pH values ranging from 3.6 to 9.5. The vibrational frequencies of the Fe-C stretching and Fe-C-O bending modes have been identified by isotopic substitution. Both His 42 --> Leu and Arg 38 --> Leu adducts with CO displayed a single Fe-C stretching band, whereas both recombinant and wild-type HRPC-CO have two bands, corresponding to different conformers. This comparison suggests that CO is H-bonded either to the distal Arg or to the distal His in the two conformers. An acid transition, common to the wild-type protein, was observed for both mutants. This indicates that these distal amino acids do not influence the acid transition. On the contrary, an alkaline transition was only observed for the Arg 38 --> Leu mutant, which suggests that distal His is involved in the alkaline transition of HRPC-CO complex. The spectroscopic information is found to be consistent with the X-ray structure of ferric HRPC. A comparison with the CO complexes of cytochrome c peroxidase and myoglobin is performed, which displays the functional significance of the structural differences between peroxidase classes I and III and between peroxidases and globins, respectively. PMID- 9753445 TI - Quantitative analysis of the effects of intrathylakoid pH and xanthophyll cycle pigments on chlorophyll a fluorescence lifetime distributions and intensity in thylakoids. AB - The xanthophyll cycle-dependent dissipation of excitation energy in higher plants is one of the most important regulatory and photoprotective mechanisms in photosynthesis. Using parallel time-resolved and pulse-amplitude modulation fluorometry, we studied the influence of the intrathylakoid pH and the xanthophyll cycle carotenoids on the PSII chlorophyll (Chl) a fluorescence yield in thylakoids of Arabidopsis, spinach, and barley. Increases in concentrations of dithiothreitol in thylakoids, which have a trans-thylakoid membrane pH gradient and are known to have decreased conversion of violaxanthin (V) to zeaxanthin (Z), lead to (1) decreases in the fractional intensity of the approximately 0.5 ns Chl a fluorescence lifetime (tau) distribution component and simultaneous increases in a 1.6-1.8 ns fluorescence component and (2) increases in the maximal fluorescence intensity. These effects disappear when the pH gradient is eliminated by the addition of nigericin. To quantitatively explain these results, we present a new mathematical model that describes the simultaneous effects of the chloroplast trans-thylakoid membrane pH gradient and xanthophyll cycle pigments on the PSII Chl a fluorescence tau distributions and intensity. The model assumes that (1) there exists a specific binding site for Z (or antheraxanthin, A) among or in an inner antenna complex (primarily CP29), (2) this binding site is activated by a low intrathylakoid pH (pK approximately 4.5) that increases the affinity for Z (or A), (3) about one Z or A molecule binds to the activated site, and (4) this binding effectively "switches" the fluorescence tau distribution of the PSII unit to a state with a decreased fluorescence tau and emission intensity (a "dimmer switch" concept). This binding is suggested to cause the formation of an exciton trap with a rapid intrinsic rate constant of heat dissipation. Statistical analysis of the data yields an equilibrium association constant, Ka, that ranges from 0.7 to 3.4 per PSII for the protonated/activated binding site for Z (or A). The model explains (1) the relative fraction of the approximately 0.5 ns fluorescence component as a function of both Z and A concentration and intrathylakoid pH, (2) the dependence of the ratio of F'm/Fm on the fraction of the 0.5 ns fluorescence tau component (where F'm and Fm are maximal fluorescence intensities in the presence and the absence of a pH gradient), and (3) the dependence of the ratio of F'm/Fm on the concentration of Z and A and the intrathylakoid pH. PMID- 9753446 TI - Characterization of the interaction between manganese and tyrosine Z in acetate inhibited photosystem II. AB - When acetate-inhibited photosystem II (PSII) membranes are illuminated at temperatures above 250 K and quickly cooled to 77 K, a 240 G-wide electron paramagnetic resonance (EPR) signal is observed at 10 K. This EPR signal arises from a reciprocal interaction between the spin 1/2 ground state of the S2 state of the Mn4 cluster, for which a multiline EPR signal with shifted 55Mn hyperfine peaks is observed, and the oxidized tyrosine residue, YZ*, for which a broadened YZ* EPR spectrum is observed. The S2YZ* EPR signal in acetate-inhibited PSII is the first in which characteristic spectral features from both paramagnets can be observed. The observation of distinct EPR signals from each of the paramagnets together with the lack of a half-field EPR transition indicates that the exchange and dipolar couplings are weak. Below 20 K, the S2YZ* EPR signal in acetate inhibited PSII is in the static limit. Above 20 K, the line width narrows dramatically as the broad low-temperature S2YZ* EPR signal is converted to a narrow YZ* EPR signal at room temperature. The line width narrowing is interpreted to be due to averaging of the exchange and dipolar interactions between YZ* and the S2 state of the Mn4 cluster by rapid spin-lattice relaxation of the Mn4 cluster as the temperature is increased. Decay of the S2YZ* intermediate at 200 K shows that the g = 4.1 form of the S2 state is formed and that a noninteracting S2-state multiline EPR signal is not observed as an intermediate in the decay. This result shows that a change in the redox state of YZ induces a spin-state change in the Mn4 cluster in acetate-inhibited PSII. The interconversion between spin states of the Mn4 cluster in acetate-inhibited PSII supports the idea that YZ oxidation or YZ* reduction is communicated to the Mn4 cluster through a direct hydrogen-bonding pathway, possibly involving a ligand bound to the Mn4 cluster. PMID- 9753447 TI - Lys75 of Anabaena ferredoxin-NADP+ reductase is a critical residue for binding ferredoxin and flavodoxin during electron transfer. AB - Previous studies, and the three-dimensional structure of Anabaena PCC 7119 ferredoxin-NADP+ reductase (FNR), indicate that the positive charge of Lys75 might be directly involved in the interaction between FNR and its protein partners, ferredoxin (Fd) and flavodoxin (Fld). To assess this possibility, this residue has been replaced by another positively charged residue, Arg, by two uncharged residues, Gln and Ser, and by a negatively charged residue, Glu. UV-vis absorption, fluorescence, and CD spectroscopies of these FNR mutants (Lys75Arg, Lys75Gln, Lys75Ser, and Lys75Glu) indicate that all the mutated proteins folded properly and that significant protein structural rearrangements did not occur. Steady-state kinetic parameters for these FNR mutants, utilizing the diaphorase activity with DCPIP, indicate that Lys75 is not a critical residue for complex formation and electron transfer (ET) between FNR and NADP+ or NADPH. However, steady-state kinetic activities requiring complex formation and ET between FNR and Fd or Fld were appreciably affected when the positive charge at position of Lys75 was removed, and the ET reaction was not even measurable if a negatively charged residue was placed at this position. These kinetic parameters also suggest that it is complex formation that is affected by mutation. Consistent with this, when dissociation constants (Kd) for FNRox-Fdox (differential spectroscopy) and FNRox-Fdrd (laser flash photolysis) were measured, it was found that neutralization of the positive charge at position 75 increased the Kd values by 50-100-fold, and that no complex formation could be detected upon introduction of a negative charge at this position. Fast transient kinetic studies also corroborated the fact that removal of the positive charge at position 75 of FNR appreciably affects the complex formation process with its protein partners but indicates that ET is still achieved in all the reactions. This study thus clearly establishes the requirement of a positive charge at position Lys75 for complex formation during ET between FNR and its physiological protein partners. The results also suggest that the interaction of this residue with its protein partners is not structurally specific, since Lys75 can still be efficiently substituted by an arginine, but is definitely charge specific. PMID- 9753448 TI - Fourier transform infrared study of the cation radical of P680 in the photosystem II reaction center: evidence for charge delocalization on the chlorophyll dimer. AB - A Fourier transform infrared (FTIR) difference spectrum of the primary electron donor (P680) of photosystem II upon its photooxidation (P680+/P680) was obtained in the frequency region of 1000-3000 cm-1. The reaction center (RC) complex (D1 D2-Cytb559) was used for the measurements in the presence of ferricyanide as an exogenous electron acceptor. Control measurements of electronic absorption (300 1200 nm) showed that illumination of the RC complex at 150 K induced major oxidation of P680 concomitant with oxidation of a carotenoid and an accessory chlorophyll (Chl). Illumination at 250 K also specifically bleached one of the two beta-carotene molecules bound to the RC complex, and the sample thus treated exhibited little formation of a carotenoid cation on subsequent illumination at 150 K. The P680+/P680 FTIR difference spectrum (with minor contamination of Chl+/Chl) was measured at 150 K using this partially carotenoid-deficient RC complex. The spectrum showed a broad positive band centered at approximately 1940 cm-1, which could be ascribed to an infrared electronic transition of P680+ analogous to that previously observed in various bacterial P+. This finding indicates that a positive charge is delocalized over (or hopping between) the two Chl molecules in P680+. The low intensity of this electronic band compared with that of the bacterial band could have three possible explanations: weak resonance interaction between the constituent Chl molecules, an asymmetric structure of P680+, and the difference in Chl species. Bands in the C=O stretching region (1600-1750 cm-1) were interpreted in comparison with resonance Raman spectra of the RC complex. The negative peaks at 1704 and 1679 cm-1 were proposed as candidates for the keto C9=O bands of P680. The observation that neither of these bands agreed with the main keto C9=O band at 1669 cm-1 in the previous 3P680/P680 FTIR spectrum [Noguchi et al. (1993) Biochemistry 32, 7186-7195] led to the idea that the triplet state migrates to a Chl (designated as ChlT) different from P680 at low temperatures. Based on these results, structural models of Chl molecules including P680 and ChlT and their coupling in the cation, triplet, and Qy singlet states are discussed. PMID- 9753449 TI - Modulation of the positional specificity of lecithin-cholesterol acyltransferase by the acyl group composition of its phosphatidylcholine substrate: role of the sn-1-acyl group. AB - Human lecithin-cholesterol acyltransferase (LCAT), which is normally specific for the sn-2 position of phosphatidylcholine (PC), derives a significant percentage of acyl groups from the sn-1 position, when sn-2 is occupied by 18:0, 20:4, or 22:6. We investigated the relative importance of the two acyl groups of PC in determining the positional specificity by first analyzing the cholesteryl esters formed in the presence of symmetric PCs labeled at sn-2. Both human and rat LCATs transferred exclusively the sn-2-acyl group from all symmetric PCs, including 18:0-18:0, and 20:4-20:4, showing that the presence of these fatty acids at sn-2 does not automatically alter the positional specificity. The role of the sn-1 acyl group was then tested by using PCs containing 20:4 or 18:0 at sn-2 and fatty acids of various chain lengths and unsaturation at sn-1. With 20:4 at sn-2 and saturated fatty acids of various chain lengths at sn-1, human and rat LCATs derived, respectively, 5-72% and 1-20% of the total acyl groups from the sn-1 position. However, the chain length of the sn-1-acyl did not correlate with its utilization by either enzyme. Various unsaturated fatty acids at sn-1 also were transferred by human LCAT at a higher rate (5-75% of total) than they were transferred by rat LCAT (0-21%). With sn-2-18:0 PCs, however, rat LCAT exhibited greater alteration in positional specificity (30-95% from sn-1) than human LCAT (15-83% from sn-1). These results show that while the primary determinant of positional specificity is the sn-2-acyl group of PC, the structure of sn-1-acyl significantly modifies it. PMID- 9753451 TI - A polar octapeptide fused to the N-terminal fusion peptide solubilizes the influenza virus HA2 subunit ectodomain. AB - As a step toward studying membrane fusion with a simplified molecule, the ectodomain, residues 1-185, of the membrane-anchored subunit HA2 of the influenza virus haemagglutinin (HA) was solubilized by adding the very polar FLAG octapeptide (Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys) to the N-terminal HA2 fusion peptide. The resulting chimeric protein, F185, when expressed in bacteria, folded spontaneously into a soluble trimer, with a high alpha-helical content and a high melting temperature, structural characteristics of the low-pH-induced conformation of HA2. Removal of the FLAG octapeptide by proteolysis with enterokinase converted the soluble molecule to one that aggregated, bound nonionic detergent, and bound to lipid vesicles, properties of the low-pH-induced conformation of HA. Thermolysin treatment of the aggregated protein removed the nonpolar fusion peptide, regenerating soluble trimers of HA2 (residues 24-185), which is analogous to thermolysin treatment of HA in the low-pH-induced conformation. Thermolysin treatment also dissociates F185 from the detergent protein complex by removing the fusion peptide. These results suggest that highly polar peptides can be fused to the membrane-binding regions of membrane proteins to increase their solubility. They also indicate that ectodomains of HA2 made in bacteria have membrane-binding properties similar to those of the same ectodomain generated by low-pH treatment of HA isolated from virus. PMID- 9753450 TI - E2P phosphoforms of Na,K-ATPase. I. Comparison of phosphointermediates formed from ATP and Pi by their reactivity toward hydroxylamine and vanadate. AB - The properties of Na,K-ATPase phosphoenzymes formed either from ATP in the presence of Mg2+ and Na+ or from Pi in the absence of alkali cations were investigated by biochemical methods and spectrofluorometry employing the styryl dye RH421. We characterized the phosphoenzyme species by their reaction to N methyl hydroxylamine, which attacks specifically the protein-phosphate bond. We studied reactions of the phospho- and dephospho-enzymes with vanadate, which is a transition-state analogue of phosphate in this enzyme. On the basis of substantial differences in the properties of the phosphoenzyme species formed either from ATP or Pi, especially in their reactivity to N-methyl hydroxylamine, it is suggested that the two phosphoenzyme species are two subconformations of the E2P phosphoform. Analysis of the RH421 fluorescence responses under a variety of experimental conditions and comparing different enzyme sources suggested that the increase of RH421 fluorescence induced by inorganic phosphate in the absence of alkali cations is associated with the formation of the covalent acyl-phosphate bond. PMID- 9753452 TI - Localization of the sites for Ca2+-binding proteins on G protein-coupled receptor kinases. AB - Inhibition of G protein-coupled receptor kinases (GRKs) by Ca2+-binding proteins has recently emerged as a general mechanism of GRK regulation. While GRK1 (rhodopsin kinase) is inhibited by the photoreceptor-specific Ca2+-binding protein recoverin, other GRKs can be inhibited by Ca2+-calmodulin. To dissect the mechanism of this inhibition at the molecular level, we localized the GRK domains involved in Ca2+-binding protein interaction using a series of GST-GRK fusion proteins. GRK1, GRK2, and GRK5, which represent the three known GRK subclasses, were each found to possess two distinct calmodulin-binding sites. These sites were localized to the N- and C-terminal regulatory regions within domains rich in positively charged and hydrophobic residues. In contrast, the unique N-terminally localized GRK1 site for recoverin had no clearly defined structural characteristics. Interestingly, while the recoverin and calmodulin-binding sites in GRK1 do not overlap, recoverin-GRK1 interaction is inhibited by calmodulin, most likely via an allosteric mechanism. Further analysis of the individual calmodulin sites in GRK5 suggests that the C-terminal site plays the major role in GRK5-calmodulin interaction. While specific mutation within the N-terminal site had no effect on calmodulin-mediated inhibition of GRK5 activity, deletion of the C-terminal site attenuated the effect of calmodulin on GRK5, and the simultaneous mutation of both sites rendered the enzyme calmodulin-insensitive. These studies provide new insight into the mechanism of Ca2+-dependent regulation of GRKs. PMID- 9753453 TI - Structural flexibility of the linker region of human P-glycoprotein permits ATP hydrolysis and drug transport. AB - P-Glycoprotein (Pgp), an energy-dependent drug efflux pump responsible for multidrug resistance of many cancer cells, is comprised of two homologous halves connected by a peptide segment approximately 75 amino acids (aa) in length. The effects of length and composition of this connecting region on Pgp cell surface expression and the ability of the two halves to interact were explored using both stable transfections of Pgp mutants in mammalian cell lines and a vaccinia virus transient expression system. A 17 aa insertion of predicted flexible structure between amino acids 681 and 682 resulted in a functional Pgp molecule that was capable of conferring drug resistance. In contrast, an 18 aa peptide insertion with a predicted alpha-helical structure was unstable when expressed transiently. A 34 aa deletion from the central core of the linker region (Delta653-686) resulted in a protein expressed at the cell surface in amounts comparable to that of wild-type Pgp but unable to confer drug resistance. No apparent differences in drug or [alpha-32P]-8-azido-ATP photoaffinity labeling were observed. However, both ATP hydrolysis and drug transport activities of the deletion mutant were completely abrogated, indicating that the linker deletion disconnected substrate binding from ATP hydrolysis and transport. This mutant also failed to exhibit an ATP hydrolysis-dependent enhancement of binding of a conformation-sensitive monoclonal antibody, UIC2. Upon replacement with a 17 aa linker peptide having a predicted flexible secondary structure, but bearing no homology to the deleted 34 aa segment, normal Pgp transport and basal and drug-stimulated ATPase activities were restored along with increased UIC2 binding in the presence of substrate, suggesting a dramatic conformational change between the nonfunctional and functional molecules. Taken together, these data suggest a flexible secondary structure of the connector region is sufficient for the coordinate functioning of the two halves of Pgp, likely specifically required for the proper interaction of the two ATP binding sites. PMID- 9753454 TI - Synthesis and biochemical characterization of an analogue of CheY-phosphate, a signal transduction protein in bacterial chemotaxis. AB - CheY is a signal transduction protein of the bacterial chemotaxis system that acts as a molecular switch to alter the swimming behavior of the bacterium. When CheY becomes phosphorylated at Asp57, CheY-Pi interacts with flagellar motor proteins, including FliM, to increase the likelihood that the flagellar motor will change its sense of rotation, increasing the frequency of tumbling. The structure of CheY in its dephosphorylated (inactive) state has been intensively investigated. The short lifetime ( approximately 20 s) of the aspartyl phosphate has precluded the complete structural determination of CheY-Pi. We have synthesized an analogue of CheY-Pi by alkylating an aspartate-to-cysteine mutant at position 57 of CheY to add a phosphonomethyl group at Cys57. This analogue, phosphono-CheY, is stable for months. Phosphono-CheY binds to two of the targets of CheY-Pi, FliM and CheZ, in a manner similar to that of CheY-Pi and much better than either unphosphorylated CheY or the unmodified form of D57C CheY. Phosphono CheY also binds Mg(II) with a dissociation constant of approximately 6 mM at neutral pH and moderate salt level. These observations indicate that phosphono CheY is a good biochemical analogue of CheY-Pi. PMID- 9753455 TI - Deamidation of specific glutamine residues from alpha-A crystallin during aging of the human lens. AB - Although it has been hypothesized that age-dependent deamidation of glutamine and/or asparagine residues may play an important role in the turnover of proteins in vivo, surprisingly little is known concerning the extents of deamidation of biologically important proteins with very long half-lives. Alpha-A crystallin is the most abundant protein of the adult human lens, which contains long-lived proteins in the central fetal-embryonic region that were synthesized before birth of the individual. Peptides, corresponding to tryptic fragments of alpha-A crystallin, were synthesized with either the expected glutamine-6, glutamine-50, and glutamine-147 residues, or deamidated glutamic acid residues at the same positions. These synthetic peptides were used to identify and quantitate the amidated versus deamidated forms of each tryptic fragment of alpha-A crystallin from the fetal-embryonic region of lenses from donors of increasing age up to 64 years old. The results demonstrate that all three glutamine residues are very stable, with glutamine-50 undergoing a maximum of approximately 30% deamidation after 64 years postsynthesis, while glutamine-6 and glutamine-147 undergo no detectable deamidation during the same period of time. Together, these results are consistent with the hypothesis that resistance to age-dependent, nonenzymatic deamidation may be an important prerequisite for the stability of proteins in vivo. PMID- 9753456 TI - Physical and functional interactions between the transactivation domain of the hematopoietic transcription factor NF-E2 and WW domains. AB - Tandem binding sites for the hematopoietic transcription factor NF-E2 in the beta globin locus control region activate high-level beta-globin gene expression in transgenic mice. NF-E2 is a heterodimer consisting of a hematopoietic subunit p45 and a ubiquitous subunit p18. Gavva et al. [Gavva, N. R., Gavva, R., Ermekova, K., Sudol, M., and Shen, J. C. (1997) J. Biol. Chem. 272, 24105-24108] reported that human p45 contains a PPXY motif that binds WW domains. We show that murine NF-E2, which contains two PPXY motifs (PPXY-1 and -2) within its transactivation domain, differentially interacted with nine GST-WW domain fusion proteins. Quantitative analysis revealed high-affinity binding (KD = 5.7 nM) of p45 to a WW domain from a novel human ubiquitin ligase homologue (WWP1) expressed in hematopoietic tissues. The amino-terminal WW domain of WWP1 formed a multimeric complex with DNA-bound NF-E2. A WWP1 ligand peptide, isolated by phage display, and a peptide spanning PPXY-1 inhibited p45 binding, whereas an SH3 domain interacting peptide and a peptide spanning PPXY-2 did not. Mutation of PPXY-1, but not PPXY-2, inhibited the transactivation function of NF-E2, providing support for the hypothesis that WW domain interactions are important for NF-E2 mediated transactivation. PMID- 9753457 TI - Iron release is reduced by mutations of lysines 206 and 296 in recombinant N terminal half-transferrin. AB - Human serum transferrin consists of two iron-binding lobes connected by a short peptide linker. While the high homology and structural similarity between the two halves of the molecule would suggest similar characteristics, it has been shown that the pH-dependent rate of release of iron from the N-terminal lobe is quite different from that of its C-terminal counterpart. This suggests that the N-lobe of human serum transferrin has a specific, pH-dependent, molecular mechanism for releasing iron. Sacchettini and co-workers using structural information have hypothesized that two lysines in the N-terminal lobe of ovotransferrin create a dilysine interaction and suggest that this is the trigger for pH-dependent iron release. To investigate this hypothesis, we used a Pichia pastoris expression system to produce large amounts of wild-type nTf, the single point mutants, nTfK206A (Lys 206 to alanine) and nTfK296A (Lys 296 to alanine), and the double mutant, nTfK206/296A. The purified recombinant proteins were then used to measure rates of iron release to pyrophosphate. It was found that the rate of iron release from all three mutant proteins at pH 5.7 (the pH at which nTf would normally release iron) was too slow to measure. Only when the pH was reduced to 5.0 could the rates of iron release from the mutant proteins be reliably determined. Although this precludes a direct comparison to wild-type nTf (the rate of iron release from nTf at pH 5.0 is too fast to measure), it implicates lysines 206 and 296 in the pH-dependent release of iron from nTf. PMID- 9753458 TI - Substitution of tyrosine for phenylalanine in fibrinopeptide A results in preferential thrombin cleavage of fibrinopeptide B from fibrinogen. AB - Phenylalanine at residue 8 in the Aalpha chain of fibrinogen is a highly conserved amino acid that is believed to be critical for binding and catalysis by the serine protease thrombin. We have examined the requirement for Phe at this position by constructing a variant recombinant fibrinogen with a conservative substitution of tyrosine for phenylalanine, Aalpha F8Y fibrinogen. We found that the variant fibrinopeptide A (F8Y 1-16) was cleaved by thrombin, in contrast to the lack of cleavage of an Aalpha 1-23 peptide and an Aalpha 1-50 fusion protein with the same substitution. This result indicates that fibrinogen residues other than Aalpha 1-50 participate in thrombin binding and fibrinogen proteolysis. We found, for the first time, that thrombin-catalyzed lysis of the fibrinogen Bbeta chain preceded lysis of the Aalpha chain, such that fibrinopeptide B (FpB) was released prior to F8Y 1-16. Kinetic analysis demonstrated that F8Y 1-16 was a very poor substrate for thrombin, with a specificity constant 280-fold lower than normal fibrinopeptide A. FpB was also a poor substrate, but the specificity constant for FpB was only 4-fold lower than normal. Consequently, FpB was preferentially released from Aalpha F8Y fibrinogen. This "role reversal" had a dramatic effect on polymerization, such that the rate of Aalpha F8Y fibrinogen polymerization was 13% of the rate of normal recombinant fibrinogen. These results confirm the importance of phenylalanine at Aalpha chain residue 8 for efficient thrombin-catalyzed proteolysis of fibrinogen, and further demonstrate that sequential fibrinopeptide release has an important role in normal polymerization. PMID- 9753459 TI - The human immunodeficiency virus type 1 Vpu protein enhances membrane permeability. AB - Infection of T lymphocytes by the human immunodeficiency virus causes drastic alterations in the intracellular cation content of the infected cells. The human immunodeficiency virus type 1 genome encodes several accessory proteins, including Vpu, an integral membrane protein that forms ion channels in planar lipid bilayers. The effect of Vpu on the permeability of the plasma membrane to several molecules has been analyzed. Expression of vpu in Escherichia coli cells increases membrane permeability to a number of molecules such as 2-nitrophenyl beta-D-galactopyranoside, uridine, the impermeable translation inhibitor hygromycin B, and lysozyme. In addition, transient expression of Vpu in eukaryotic COS cells enhances entry of charged molecules such as hygromycin B and neurobiotin into these cells. The effect of Vpu on cell membrane permeability resembles that reported for other membrane-active proteins from different animal viruses, including influenza M2, Semliki Forest virus 6K, and poliovirus 2B and 3A proteins. PMID- 9753460 TI - The very low- and intermediate-density lipoprotein fraction isolated from apolipoprotein E-knockout mice transforms macrophages to foam cells through an apolipoprotein E-independent pathway. AB - Apolipoprotein E (apoE)-knockout mice develop severe atherosclerosis associated with high levels of very low-density lipoprotein (VLDL) and intermediate-density lipoprotein (IDL) in plasma. To investigate the atherogenic role of VLDL and IDL, the lipoprotein fraction containing both VLDL and IDL (apoEko-VLDL/IDL) was isolated from plasma of apoE-knockout mice by ultracentrifugation, and its interaction with macrophages was studied. When peritoneal macrophages obtained from apoE-knockout mice were incubated with apoEko-VLDL/IDL, the level of cellular cholesteryl esters (CE) increased with the concentration of apoEko VLDL/IDL. The level of cellular cholesteryl [3H]oleate formed reached 15.1 nmol/mg of cell protein upon incubation with 50 microg/mL apoEko-VLDL/IDL for 18 h, which was an 8.4-fold increase over the corresponding level induced by low density lipoprotein (LDL). The cellular CE mass was also significantly increased by apoEko-VLDL/IDL. Morphologically, after exposure to apoEko-VLDL/IDL, macrophages became strongly stained with Sudan black B. The total binding of [125I]apoEko-VLDL/IDL to macrophages was effectively replaced by more than 80% by an excess of the unlabeled ligand. Specific binding, calculated by subtracting the nonspecific binding from the total binding, exhibited a saturation pattern. Similar results were obtained with cell association and degradation experiments. In addition, the endocytic degradation of [125I]apoEko-VLDL/IDL was partially inhibited by LDL, whereas acetyl-LDL did not show any effect. These results indicated that apoEko-VLDL/IDL in its unmodified form produced significant CE accumulation in macrophages through a specific and apoE-independent pathway. This pathway may explain, in part, the mechanisms of foam cell formation in arterial walls and the subsequent development of atherosclerosis in apoE-knockout mice. PMID- 9753461 TI - Backbone flexibility of five sites on the catalytic subunit of cAMP-dependent protein kinase in the open and closed conformations. AB - To develop an alternative approach to measure peptidyl backbone flexibility and to expand our understanding of the segmental flexibility of cAMP-dependent protein kinase (cAPK), the effect of protein kinase inhibitor peptide, PKIalpha(5 24), and MgATP on the mobility of fluorescein selectively conjugated to five sites on the catalytic subunit of cAPK was examined. Specifically, five full length, single-site catalytic subunit mutants (K16C, K81C, I244C, C199A, and N326C) were prepared, and fluorescein maleimide was selectively attached to the side chains of each substituted cysteine or, in the case of the C199A mutant, to the unprotected native C343. The time-resolved anisotropy decay profiles of the five fluorescein maleimide-conjugated mutants were well fit to a biexponential equation. The fast rotational correlation times of the fluorescein conjugates ranged between 1.9 and 2.8 ns and were inversely correlated (r = -0.87) to the averaged crystallographic main-chain atom B factors around each site of conjugation. The slow correlation times ranged between 25 and 28 ns and were about the same magnitude as the value of 21 ns estimated from the Stokes-Einstein equation. The presence of MgATP and PKIalpha(5-24), which induces the closed conformation of cAPK, was associated with a reduction of the fast rotational correlation time of the K81C conjugate, indicating that the peptidyl backbone around K81 is measurably less flexible when the C subunit is in the closed compared with the open conformation. The results suggest (i) that time-resolved fluorescence anisotropy can assess the nanosecond flexibility of short segments of the peptidyl backbone around each site of labeling and (ii) that the substrate/pseudosubstrate binding differentially affects the backbone flexibility of cAPK. PMID- 9753462 TI - Effects of secondary structure on DNA and RNA cleavage by diplatinum(II). AB - The photochemistry of Pt2(pop)44- with nucleic acids has been studied using radiolabeled oligomers of DNA and RNA and high-resolution electrophoresis (pop is P2O5H22-). Photolysis of Pt2(pop)44- with duplex DNA produces an even cleavage ladder and relatively little enhancement of cleavage upon treatment with piperidine. In contrast, the cleavage pattern is far less regular with single stranded DNA, and there is a large enhancement in cleavage upon treatment with piperidine. Accordingly, photolysis of Pt2(pop)44- with the DNA hairpin 5' d[ATCCTATTTATAGGAT] produces a much larger piperidine enhancement at the loop and end nucleotides than in the stem. There is an additional piperidine enhancement that occurs selectively at guanine residues either in RNA or in DNA at low Mg2+ concentrations that is attributed to outer-sphere electron transfer on the basis of the known excited-state redox potentials of Pt2(pop)44- and the expected oxidative chemistry of guanine. The extent of guanine oxidation is higher compared to the extent of sugar oxidation at low Mg2+ concentrations, which can be attributed to a shallower distance dependence for electron transfer compared to that for atom transfer. The effects of Mg2+ and piperidine or aniline treatment were examined on stem-loop structures of DNA and RNA and gave partial images of the expected secondary structures. PMID- 9753463 TI - Ca2+ binding and conformational changes in a calmodulin domain. AB - Calcium activation of the C-terminal domain of calmodulin was studied using 1H and 15N NMR spectroscopy. The important role played by the conserved bidentate glutamate Ca2+ ligand in the binding loops is emphasized by the striking effects resulting from a mutation of this glutamic acid to a glutamine, i.e. E104Q in loop III and E140Q in loop IV. The study involves determination of Ca2+ binding constants, assignments, and structural characterizations of the apo, (Ca2+)1, and (Ca2+)2 states of the E104Q mutant and comparisons to the wild-type protein and the E140Q mutant [Evenas et al. (1997) Biochemistry 36, 3448-3457]. NMR titration data show sequential Ca2+ binding in the E104Q mutant. The first Ca2+ binds to loop IV and the second to loop III, which is the order reverse to that observed for the E140Q mutant. In both mutants, the major structural changes occur upon Ca2+ binding to loop IV, which implies a different response to Ca2+ binding in the N- and C-terminal EF-hands. Spectral characteristics show that the (Ca2+)1 and (Ca2+)2 states of the E104Q mutant undergo global exchange on a 10-100 micros time scale between conformations seemingly similar to the closed and open structures of this domain in wild-type calmodulin, paralleling earlier observations for the (Ca2+)2 state of the E140Q mutant, indicating that both glutamic acid residues, E104 and E140, are required for stabilization of the open conformation in the (Ca2+)2 state. To verify that the NOE constraints cannot be fulfilled in a single structure, solution structures of the (Ca2+)2 state of the E104Q mutant are calculated. Within the ensemble of structures the precision is good. However, the clearly dynamic nature of the state, a large number of violated distance restraints, ill-defined secondary structural elements, and comparisons to the structures of calmodulin indicate that the ensemble does not provide a good picture of the (Ca2+)2 state of the E104Q mutant but rather represents the distance-averaged structure of at least two distinct different conformations. PMID- 9753464 TI - Phosphorylation of the C-terminal sites of human p53 reduces non-sequence specific DNA binding as modeled with synthetic peptides. AB - Phosphorylation of the tumor suppressor p53 is generally thought to modify the properties of the protein in four of its five independent domains. We used synthetic peptides to directly study the effects of phosphorylation on the non sequence-specific DNA binding and conformation of the C-terminal, basic domain. The peptides corresponded to amino acids 361-393 and were either nonphosphorylated or phosphorylated at the protein kinase C (PKC) site, Ser378, or the casein kinase II (CKII) site, Ser392, or bis-phosphorylated on both the PKC and the CKII sites. A fluorescence polarization analysis revealed that either the recombinant p53 protein or the synthetic peptides bound to two unrelated target DNA fragments. Phosphorylation of the peptide at the PKC or the CKII sites clearly decreased DNA binding, and addition of a second phosphate group almost completely abolished binding. Circular dichroism spectroscopy showed that the peptides assumed identical unordered structures in aqueous solutions. The unmodified peptide, unlike the Ser378 phosphorylated peptide, changed conformation in the presence of DNA. The inherent ability of the peptides to form an alpha-helix could be detected when circular dichroism and nuclear magnetic resonance spectra were taken in trifluoroethanol-water mixtures. A single or double phosphorylation destabilized the helix around the phosphorylated Ser378 residue but stabilized the helix downstream in the sequence. PMID- 9753465 TI - Benzoquinazoline derivatives as substitutes for thymine in nucleic acid complexes. Use of fluorescence emission of benzo[g]quinazoline-2,4-(1H,3H)-dione in probing duplex and triplex formation. AB - Triple helix formation obeys structural features that do not allow accommodation of every double-stranded sequence; it requires the occurrence of homopurine stretches. A further constraint comes from the weak energy of interaction between the third strand and the double-stranded target. In an attempt to design bases leading to increased stability of triplexes, we explored the ability of modified bases with an extended aromatic domain to increase third strand binding through stacking interactions. We report here the use of benzo[g]- and benzo[f]quinazoline-2,4-dione-(1H,3H)-dione as substitutes for thymine in the canonical TAT triplet. The synthesis and characterization of the beta nucleoside derivatives of benzoquinazolines are described. Triplex-forming oligonucleotides containing these modified bases have been prepared, and their ability to form triplexes has been evaluated by UV absorption-monitored thermal denaturation measurements. Benzo[g]quinazoline and benzo[f]quinazoline formed triple-stranded structures with slightly decreased stabilities. In addition, benzo[g]quinazoline revealed strong fluorescence emission properties which can be used to monitor selectively the formation of triple-helical structures. Annealing of benzo[g]quinazoline to complementary strands did not produce any fluorescence modification. But when it was introduced into the Hoogsteen strand of PyPuPy complexes, the fluorescence intensity was reduced and the emission maximum was shifted to short wavelengths. PMID- 9753466 TI - A single mutation Asp229 --> Ser confers upon Gs alpha the ability to interact with regulators of G protein signaling. AB - RGS proteins (regulators of G protein signaling) are GTPase activating proteins (GAPs) for Gi and Gq families of heterotrimeric G proteins but have not been found to interact with Gs alpha. The Gs alpha residue Asp229 has been suggested to be responsible for the inability of RGS proteins to interact with Gs alpha [Natochin, M., and Artemyev, N. O. (1998) J. Biol. Chem. 273, 4300-4303]. To test this hypothesis, we have investigated the possibility of generating an interaction between Gs alpha and RGS proteins by substituting Gs alpha Asp229 with Ser and replacing the potential Gs alpha Asp229 contact residues in RGS16, Glu129 and Asn131, by Ala and Ser, respectively. RGS16 and its mutants failed to interact with Gs alpha. A single mutation of Gs alpha, Asp229Ser, rendered the Gs alpha subunit with the ability to interact with RGS16 and RGS4. Like RGS protein binding to Gi and Gq alpha-subunits, RGS16 preferentially recognized the AlF4- bound conformation of Gs alpha Asp229Ser. In a single-turnover assay, RGS16 maximally stimulated GTPase activity of Gs alpha Asp229Ser by approximately 5 fold with an EC50 value of 7.5 microM. Our findings demonstrate that Asp229 of Gs alpha represents a major barrier for Gs alpha interaction with known RGS proteins. PMID- 9753467 TI - Random versus selective membrane phospholipid oxidation in apoptosis: role of phosphatidylserine. AB - The formation of reactive oxygen species has been associated with apoptosis. To assess the role of lipid peroxidation in apoptosis, we used 2,2'-azobis(2,4 dimethylisovaleronitrile) (AMVN) to generate peroxyl radicals within cellular membranes of HL-60 cells. cis-Parinaric acid (cis-PnA) metabolically integrated into phospholipids of HL-60 cells was used as a probe to assess the extent of lipid peroxidation within specific phospholipid classes. Within 2 h, AMVN (500 microM) randomly oxidized more than 85% of cis-PnA contained in all major classes of phospholipids. AMVN-induced lipid peroxidation was followed by apoptosis as determined by nuclear condensation, DNA fragmentation, and annexin V binding to externalized phosphatidylserine (PS). Fluorescamine derivatization of external aminophospholipids revealed that PS, but not phosphatidylethanolamine, was externalized. The vitamin E analogue, 6-hydroxy-2,2,5,7,8-pentamethylchromane (PMC), inhibited overall oxidation of cis-PnA in phospholipids by more than 85%. Not all phospholipids, however, were equally protected. Phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, and sphingomyelin were nearly completely protected by PMC, while oxidation of PS was unaffected in whole living cells. The insensitivity of PS to PMC was not an intrinsic property because PMC protected all lipids equally during AMVN oxidation of liposomes prepared from cis PnA-labeled cells. The potential role for PS oxidation in apoptosis was further suggested by the faithful execution of apoptosis following coexposure of cells to AMVN and PMC. PMID- 9753468 TI - Insertion of magainin into the lipid bilayer detected using lipid photolabels. AB - We investigated the interaction of the antimicrobial peptides Ala19-magainin 2 amide and magainin 2 amide with lipid using two lipid photolabels, azidobenzoyl galactosylceramide (GalCer-PL) and azidobenzoylamido capryloyl galactosylceramide (GalCer-C8-PL), which position their photosensitive groups near the apolar-polar interface and near the center of the bilayer, respectively. Magainins have been postulated to permeabilize membranes either by inserting in a transmembrane fashion into the bilayer and forming a channel or by binding to the surface of the bilayer and disturbing lipid packing. Evidence for channel formation has been difficult to obtain, possibly because only a fraction of the peptide may form a channel at any one time and because the channels may have a short lifetime. Both photolabels significantly labeled the peptides when bound to acidic phospholipid vesicles. The extent of labeling by GalCer-C8-PL was at least 70% of that by GalCer-PL, indicating that some of the peptide was inserted deeply into the bilayer at least transiently. The extent of labeling of Ala19-magainin 2 amide increased significantly with an increase in the peptide to lipid mole ratio, indicating cooperativity and supporting the channel model. The extent of labeling of this peptide was maximal by 30 s and did not change over 30 min, indicating that peptide insertion is rapid and either that the peptide remains inserted for at least 30 min or that equilibrium between inserted and noninserted peptide is achieved by 30 s. The latter is supported by other studies in the literature. Use of this hydrophobic photolabeling technique has permitted detection of peptide monomers which inserted into the bilayer and/or formed a channel at some time during the labeling procedure. PMID- 9753469 TI - A conserved aspartate residue, Asp187, is important for Na+-dependent proline binding and transport by the Na+/proline transporter of Escherichia coli. AB - Asp187 in the Na+/proline transporter of Escherichia coli (PutP) is conserved within the Na+/solute cotransporter family. Information on the role of this residue has been gained by amino acid substitution analysis. PutP with Glu, Asn, or Cys in place of Asp187 catalyzed Na+-coupled proline uptake at 75%, 25%, and 1.5%, respectively, of the Vmax of PutP-wild-type while the apparent Km for proline was only slightly altered. Importantly, acetylation or amidoacetylation of an engineered transporter containing a single Cys at position 187 stimulated proline uptake. Strikingly, PutP-D187C exhibited high-affinity proline binding even at very low Na+ concentrations (2 microM) while proline binding to PutP-wild type, -D187E, and -D187N was strictly dependent on the Na+ concentration. The apparent independence of proline binding from the Na+ concentration can at least partially be attributed to an enhanced Na+ affinity of PutP-D187C. In addition, reaction of PutP containing a single Cys at position 187 with N-ethylmaleimide was inhibited by Na+ but not by Li+ or proline. The results indicate that electrostatic interactions of the amino acid side chain at position 187 in PutP with other parts of the transporter and/or the coupling ion are crucial for active proline transport. It is suggested that Asp187 is located close to the pathway of the coupling ion through the membrane and may be involved in the release of Na+ on the cytoplasmic side of the membrane. PMID- 9753470 TI - Translesional synthesis on DNA templates containing an estrogen quinone-derived adduct: N2-(2-hydroxyestron-6-yl)-2'-deoxyguanosine and N6-(2-hydroxyestron-6-yl) 2'-deoxyadenosine. AB - Miscoding properties induced by estrogen quinone-derived DNA adducts were analyzed using an in vitro experimental system to quantify base substitutions and deletions. Site-specifically modified oligodeoxynucleotides containing a single N2-(2-hydroxyestron-6-yl)-2'-deoxyguanosine (2-OHE1-N2-dG) or N6-(2-hydroxyestron 6-yl)-2'-deoxyadenosine (2-OHE1-N6-dA) were prepared postsynthetically and used as templates in primer extension reactions catalyzed by mammalian DNA polymerases (pol) alpha, beta, and delta. The 2-OHE1-N2-dG adduct blocked primer extension reactions more strongly than 2-OHE1-N6-dA. Using pol alpha and delta, 2-OHE1-N2 dG promoted incorporation of dCMP (6.3 and 3.1%, respectively), the correct base, opposite the lesion: when pol delta was used, misincorporation of dTMP (0.52%) was detected. 2-OHE1-N6-dA also promoted incorporation of dTMP, the correct base, opposite the lesion, accompanied by misincorporation of dCTP (0.54% for pol alpha and 3.2% for pol delta) and one-base deletion (0.3-0.5%). Using pol beta, no miscoding was detected. The miscoding occurred only when replicative DNA polymerases were used. Kinetic data were consistent with those obtained from the analysis of fully extended products formed by pol alpha or pol beta. These results indicate that endogenous estrogen quinone-derived DNA adducts have miscoding potential: G --> A and A --> G transitions and deletions are predicted in mammalian cells. PMID- 9753471 TI - Identification and requirement of three ribosome binding domains in dsRNA dependent protein kinase (PKR). AB - The interferon-inducible, double-stranded (ds) RNA-dependent protein kinase (PKR) regulates protein synthesis initiation by phosphorylating the alpha-subunit of eukaryotic translation initiation factor 2 (eIF-2). The amino-terminal half of PKR contains two dsRNA binding domains, and the kinase domain resides in the carboxy-terminal half of the protein. PKR is a ribosomal-associated protein. In this report, we provide evidence that PKR contains three ribosome interaction sites, two that are localized in each of the dsRNA binding domains and one that is localized in the kinase domain. All three domains can associate with polysomes independently. The ribosome association of the dsRNA binding domains requires dsRNA binding activity. Ribosome interaction of either the individual or the combined dsRNA binding domains was disrupted by 0.1 M KCl. In contrast, the ribosome interaction of intact PKR and the isolated kinase domain was largely resistant to 0.5 M KCl. These results indicate that all three domains of PKR contribute to the high-affinity ribosomal association. After dissociation of polysomes with EDTA, both intact PKR and the isolated kinase domain were primarily associated with the 60S ribosomal subunit. Coexpression of the adenovirus VAI RNA, an RNA polymerase III gene product that binds and inactivates PKR, disrupted ribosomal association of intact PKR, but not of the isolated PKR kinase domain. The results support a model where VAI RNA induces a major conformational change in PKR to prohibit ribosome association of all interaction sites. In contrast, other inhibitors of PKR including vaccinia virus E3L and K3L gene products, and the HIV trans-activating response (TAR) element binding protein TRBP, did not disrupt ribosome association of PKR. The results suggest a novel mechanism by which viral RNAs may inactivate PKR through disrupting ribosome association. PMID- 9753472 TI - Artificial nine zinc-finger peptide with 30 base pair binding sites. AB - It is well-known that DNA binding of native nine zinc-finger protein TEIIIA is dominated by interaction of select few fingers. Newly designed zinc-finger peptide Sp1ZF9 containing nine Cys2-His2 type motifs has been manipulated. The DNA-binding property of Sp1ZF9 was compared with those of native three zinc finger Sp1(530-623) and artificial six zinc-finger Sp1ZF6 peptides. Although the equilibrium time was less than 0.5 h for Sp1(530-623)-DNA complex, Sp1ZF6 and Sp1ZF9 required approximately 48 and 72 h, respectively, for full complex formation. Evidently, the footprinting analysis demonstrated that Sp1ZF9 and Sp1ZF6 bind at least 27 and 18 contiguous base pairs of DNA sequence, respectively. Sp1ZF9 showed two step bindings to DNA, namely first the recognition of GC (5'-GGG-GCG-GGGCC-3') sequence by the N-terminal Sp1 domain and next the recognition of the corresponding target sequences by the middle and C terminal Sp1 domains. In contrast with unimolecular binding of Sp1ZF9 and Sp1ZF6, two Sp1(530-623) molecules bind to one GCIII (5'-GGG-GCG-GGG-GGG-GCG-GGG-GGG++ + GCG-GGGCC-3') site region. Semispecific complex formed at the beginning of Sp1ZF9 DNA interaction has also been characterized by kinetic analysis using surface plasmon resonance. Interestingly, the association rate constants for GC and GCIII complexes of Sp1ZF9 are smaller than those of the corresponding Sp1(530-623) complexes. Of special interest is the fact that new nine zinc-finger peptide Sp1ZF9 can bind to DNA sequence of approximately 30 base pairs. Such multi zinc finger peptides may be useful as genome-specific transcriptional switches in future. PMID- 9753473 TI - Drug-resistant HIV-1 proteases identify enzyme residues important for substrate selection and catalytic rate. AB - A series of mutations, first identified in protease inhibitor-resistant HIV-1 viral isolates, were introduced into HIV-1 PR as individual substitutions. Mutants containing R8K, V32I, V82T, I84V, G48V/L90M, or V82T/I84V substitutions were analyzed for differences in substrate preference and catalytic efficiency using a set of single amino acid substituted HIV-1 CA-NCa cleavage site peptides. All mutants exhibited wild-type preference for large hydrophobic residues, especially Phe, in the P1' substrate position. Only the R8K and V32I mutants showed significant differences in subsite selection compared to wild-type enzyme. In a parallel study, the individual mutations R10K, L12V, I44V, A60M, I71V, and I108V were introduced into RSV PR. These amino acid positions are structurally equivalent to Arg8, Leu10, Val32, Met46, Ile54, and Ile84 in HIV-1 PR, respectively, which mutate in drug-resistance. The RSV R10K substitution significantly altered substrate specificity and catalytic rate, compared to wild type, in a manner similar to that of the HIV-1 R8K mutant. Crystal structures of the RSV PR R10K, I44V, I71V, and Il08V mutant enzymes presented here indicate that each of these substitutions has little effect on the overall structure of the respective enzymes. Taken together, these data provide an explanation for the reported in vivo predilection for selection of large hydrophobic residues in the P1' substrate position of second locus mutations in the Gag polyprotein PR cleavage sites. The data also suggest that the selection of resistant enzymes is not simply limited to loss of binding to inhibitor but affects other steps in proteolysis. PMID- 9753474 TI - The activation state of p38 mitogen-activated protein kinase determines the efficiency of ATP competition for pyridinylimidazole inhibitor binding. AB - The serine/threonine kinase p38 is a ubiquitous, highly conserved, stress responsive, signal-transducing enzyme. It regulates the production of proinflammatory mediators and is the target of the cytokine synthesis inhibitory pyridinylimidazoles. We have expressed human p38 in Drosophila S2 cells and characterized preparations of mixed unphosphorylated/monophosphorylated (inactive) and homogeneously diphosphorylated (active) forms of the enzyme. We observed that only the active preparation of the enzyme has significant kinase activity when assayed using an ATF2-GST fusion protein as the substrate. We determined that the value of KM[ATP] in this reaction is 25 microM and that the pyridinylimidazole inhibitor of p38 kinase activity, SB203580, competes with ATP. We have found that a tritiated pyridinylimidazole, SB202190, has an equal affinity for both the active and inactive forms of the enzyme and that SB203580 competes with it equally well for binding to either form of the enzyme. However, ATP can compete with the tritiated inhibitor for binding to only the active form of the enzyme. Further, we demonstrate in vivo that at concentrations consistent with its IC50 as a cytokine inhibitor, SB203580 can inhibit stimulus-induced phosphorylation of p38 at the Thr-Gly-Tyr activation motif. Our observations suggest that pyridinylimidazoles may block the biological activity of p38 kinase by binding to the inactive form of p38 and reducing its rate of activation. Under these conditions, ATP would not effectively compete with the inhibitors in vivo. PMID- 9753475 TI - On the mechanism and specificity of soluble, quinoprotein glucose dehydrogenase in the oxidation of aldose sugars. AB - Kinetic and optical studies were performed on the reductive half-reaction of soluble, quinoprotein glucose dehydrogenase (sGDH), i.e., on the conversion of sGDHox plus aldose sugar into sGDHred plus corresponding aldonolactone. It appears that the nature and stereochemical configuration of the substituents at certain positions in the aldose molecule determine the substrate specificity pattern: absolute specificity exists with respect to the C1-position (only sugars being oxidized which have the same configuration of the H/OH substituents at this site as the beta-anomer of glucose, not those with the opposite one) and with respect to the overall conformation of the sugar molecule (sugars with a 4C1 chair conformation are substrates, those with a 1C4 one are not); the nature and configuration of the substituents at the 3-position are hardly relevant for activity, and an equatorial pyranose group at the 4-position exhibits only aspecific hindering of the binding of the aldose moiety of a disaccharide. The pH optimum determined for glucose oxidation appeared to be 7.0, implying that reoxidation of sGDHred is rate-limiting with those electron acceptors displaying a different value under steady-state conditions. The kinetic mechanism of sGDH consists of (a) step(s) in which a fluorescing intermediate is formed, and a subsequent, irreversible step, determining the overall rate of the reductive half reaction. The consequences of this for the likeliness of chemical mechanisms where glucose is oxidized by covalent catalysis in which a C5-adduct of glucose and PQQ are involved, or by hydride transfer from glucose to PQQ, followed by tautomerization of C5-reduced PQQ to PQQH2, are discussed. The negative cooperative behavior of sGDH seems to be due to substrate-occupation-dependent subunit interaction in the dimeric enzyme molecule, leading to a large increase of the turnover rate under saturating conditions. PMID- 9753476 TI - Conformational changes of an Hsp70 molecular chaperone induced by nucleotides, polypeptides, and N-ethylmaleimide. AB - Hsp70 molecular chaperones are highly conserved ATPases that guide the folding and assembly of proteins in many cellular pathways. They use the energy of ATP binding and hydrolysis to regulate their interactions with hydrophobic regions of unfolded proteins. The activities and the conformations of the N-terminal nucleotide- and C-terminal polypeptide-binding domains of Hsp70s are coupled. We recently reported that the sulfhydryl-modifying reagent N-ethylmaleimide (NEM) inactivates the yeast Hsp70 Ssa1p by reacting with its three cysteine residues which are located in the nucleotide-binding domain. To further characterize conformational changes associated with interdomain coupling and to determine whether NEM alters Ssa1p's conformation, the structures of Ssa1p and NEM-modified Ssa1p (NEM-Ssa1p) were compared using a variety of biophysical techniques. Size exclusion chromatography revealed that NEM-Ssa1p is more oligomeric and more resistant to nucleotide- or polypeptide-dependent depolymerization than Ssa1p. Measurement of the thermal stability indicated that NEM modification has an effect very similar to that of binding of nucleotides to the unmodified protein. Circular dichroism demonstrated small differences in the secondary structure of Ssa1p and NEM-Ssa1p, and in their complexes with nucleotides. NEM modification increased the ANS fluorescence of Ssa1p and exposed numerous trypsin-sensitive sites in its nucleotide-binding domain. The intrinsic fluorescence of Ssa1p's only tryptophan residue, which is located in a C-terminal alpha-helical region adjacent to the polypeptide-binding cleft, was quenched in the presence of ATP, but not ADP. NEM modification altered nucleotide-dependent changes in the intrinsic fluorescence of Ssa1p. Together, these results demonstrate that NEM alters the conformation of Ssa1p and disrupts, but does not eliminate, interdomain communication. Furthermore, the results provide evidence for a model in which the polypeptide-binding cleft of Hsp70s is covered by an alpha-helical lid that is open in the presence of ATP, but closed in the presence of ADP. PMID- 9753477 TI - The oligomycin sensitivity conferring protein of rat liver mitochondrial ATP synthase: arginine 94 is important for the binding of OSCP to F1. AB - The oligomycin sensitivity conferring protein (OSCP) is an essential subunit of the mitochondrial ATP synthase (F0F1) long regarded as being directly involved in the energetic coupling of proton transport to ATP synthesis. To gain insight into the function of OSCP, mutations were made in a highly conserved central region of the subunit, and the recombinant proteins were studied using several biochemical assays. Rat liver OSCP was expressed to high levels in Escherichia coli, solubilized from inclusion bodies, renatured, and purified to homogeneity. The recombinant protein was able to reconstitute oligomycin-sensitive ATPase activity to inner membrane vesicles depleted of F1 and OSCP, and bound to F1 with a stoichiometry of 1:1. A novel fluorescence anisotropy assay was developed to study the affinity of binding of F1 to OSCP, providing a Kd value of 51 +/- 11 nM. Two highly conserved, charged residues (E91 and R94) which lie within the central region of OSCP were mutated, and the recombinant proteins (E91Q, R94Q, and R94A) were purified to homogeneity and judged by CD spectroscopy to have structures similar to that of the wild-type protein. Both R94 mutants demonstrated little or no binding to F1, while the E91Q bound in a manner identical to that of wild-type OSCP. Significantly, all three mutant proteins were able to reconstitute F1 with membranes and to confer oligomycin sensitivity to the same extent as wild-type OSCP. These results demonstrate that a single tight binding site exists on isolated rat liver F1 for OSCP, and implicate arginine 94 as playing a critical role in this site. In addition, these results indicate that this tight binding site is not required for conferral of oligomycin sensitivity to the reconstituted F0F1 complex. PMID- 9753478 TI - Vibrational spectrum associated with the reduction of tyrosyl radical D* in photosystem II: a comparative biochemical and kinetic study. AB - Photosystem II (PSII) contains a redox-active tyrosine, D. Difference FT-IR spectroscopy can be used to obtain structural information about this species, which is a neutral radical, D*, in the photooxidized form. Previously, we have used isotopic labeling, site-directed mutagenesis, and kinetics to assign a vibrational line at 1477 cm-1 to D*; these studies were performed on highly resolved PSII preparations at pH 7.5 ?Kim et al. (1998) Biochim. Biophys. Acta 1364, 337-360; publisher's correction, Biochim. Biophys. Acta 1366, 330-354?. Here, we use kinetics to assign vibrational features to tyrosyl radical, D*, in PSII membranes. EPR and fluorescence controls identify a time regime in which D* decay occurs independently of redox changes involving the PSII quinone acceptors. Difference FT-IR spectra, acquired over this time regime, exhibit decreases in the amplitude of a 1477 cm-1 line; quantitative comparison with EPR transients supports the assignment to D*. Conditions, requiring the use of phosphate/formate, have been described for observation of a dissimilar FT-IR spectrum, which has been assigned to tyrosyl radical D*; this spectrum lacks a 1477 cm-1 line ?Hienerwadel et al. (1997) Biochemistry 36, 14712-14723?. Under these conditions, we have observed (1) an acceleration in the rate of D* decay and a decrease in D* yield attributable to the presence of formate, (2) a proportional decrease in the amplitude of FT-IR spectra acquired over the time regime in which D* decays, (3) frequency shifts in the D* - D FT-IR spectrum, (4) large-scale structural changes, as assessed by the amide I line shape, and (5) contributions to the FT-IR spectrum from the phosphate/formate buffer in the absence of PSII. We conclude that changes in the FT-IR spectrum, observed in the presence of phosphate/formate, are caused by alterations in the environment of D* and by direct phosphate/formate contributions to the spectrum. PMID- 9753479 TI - D221 in thymidylate synthase controls conformation change, and thereby opening of the imidazolidine. AB - In thymidylate synthase (TS), the invariant residue Asp-221 provides the only side chain that hydrogen bonds to the pterin ring of the cofactor, 5,10-methylene 5,6,7,8-tetrahydrofolate. All mutants of D221 except cysteine abolish activity. We have determined the crystal structures of two ternary complexes of the Escherichia coli mutant D221N. In a complex with dUMP and the antifolate 10 propargyl-5,8-dideazafolate (CB3717), dUMP is covalently bound to the active site cysteine, as usual. CB3717, which has no imidazolidine ring, is also bound in the usual productive orientation, but is less ordered than in wild-type complexes. The side chain of Asn-221 still hydrogen bonds to N3 of the quinazoline ring of CB3717, which must be in the enol form. In contrast, the structure of D221N with 5-fluoro-dUMP and 5,10-methylene-5,6,7, 8-tetrahydrofolate shows the cofactor bound in two partially occupied, nonproductive binding sites. In both binding modes, the cofactor has a closed imidazolidine ring and adopts the solution conformation of the unbound cofactor. In one of the binding sites, the pterin ring is turned around such that Asn-221 hydrogen bonds to the unprotonated N1 instead of the protonated N3 of the cofactor. This orientation blocks the conformational change required for forming covalent ternary complexes. Taken together, the two crystal structures suggest that the hydrogen bond between the side chain of Asp-221 and N3 of the cofactor is most critical during the early steps of cofactor binding, where it enforces the correct orientation of the pterin ring. Proper orientation of the cofactor appears to be a prerequisite for opening the imidazolidine ring prior to formation of the covalent steady-state intermediate in catalysis. PMID- 9753481 TI - Anion activation of 3-phosphoglycerate kinase requires domain closure PMID- 9753480 TI - Importance of central alpha-helices of human apolipoprotein A-I in the maturation of high-density lipoproteins. AB - We have studied the role of amphipathic alpha-helices in the ability of apoA-I to promote cholesterol efflux from human skin fibroblasts and activate lecithin:cholesterol acyltransferase (LCAT). Three apoA-I mutants were designed, each by deletion of a pair of predicted adjacent central alpha-helices [Delta(100 143), Delta(122-165), Delta(144-186)], and expressed in Escherichia coli. This strategy was used to minimize disruption of the predicted secondary structure of the resulting protein. These three central deletion mutants have been previously shown to be expressed as stable folded proteins but to exhibit altered phospholipid-binding properties. When recombined with phospholipids to form homogeneous LpA-I containing equivalent amounts of POPC and tested for their ability to promote diffusional cholesterol efflux from normal [3H]cholesterol labeled fibroblasts, each mutant and the wild-type recombinant protein (Rec.-apoA I) promoted cholesterol efflux with very similar rates at all the concentrations tested. These experiments showed that all LpA-I could acquire cellular cholesterol with similar affinity and binding capacity. However, when the cell incubated LpA-I were incubated with purified LCAT, two mutants, Delta(122-165) and Delta(144-186), appeared incapable of activating the enzyme. To directly determine their ability to activate LCAT, each mutant and the control were recombined with equivalent amounts of cholesterol and phospholipid and incubated with the purified enzyme. The results show that whereas deletion of residues 100 143 has little effect on LCAT activation, deletion of residues 122-165 or 144-186 results in an inability of the mutants to promote cholesterol esterification. In conclusion, our results show that no specific sequence in the central domain of apoA-I is required for efficient diffusional cholesterol efflux from normal fibroblasts; however, residues 144-186 appear critical for optimum LCAT activation and cholesteryl ester accumulation. Since deletion of residues 144-186 also perturbs phospholipid association and prevents the formation of large LpA-I particles [Frank, P. G., Bergeron, J., Emmanuel, F., Lavigne, J. P., Sparks, D. L., Denefle, P., Rassart, E., and Marcel, Y. L. (1997) Biochemistry 36, 1798 1806], the data show that this pair of alpha-helices plays an important role in the maturation of HDL. Sequence analysis of these apoA-I helices further identifies specific residues that appear essential to this activity. PMID- 9753482 TI - Safety of azathioprine and 6-mercaptopurine in pediatric patients with inflammatory bowel disease. AB - BACKGROUND & AIMS: Azathioprine (AZA) and 6-mercaptopurine (6-MP) are used in pediatric patients with ulcerative colitis and Crohn's disease to reduce disease activity, maintain remission, prevent relapse, and lower corticosteroid dosage, but their long-term side effects remain to be studied. The aim of this study was to analyze the safety of AZA and 6-MP and steroid reduction in this age group. METHODS: The investigators' database identified 118 patients who received either drug; 23 were excluded (single visit, noncompliance, or therapy < 1 week), leaving 95 patients, with a mean (+/-SD) age of 14.2 +/- 4.4 years. Medical files were reviewed for adverse side effects: fever, pancreatitis, infections, gastrointestinal intolerance, aminotransferase level increase, leukopenia, and thrombocytopenia. Prednisone doses before and after immunomodulatory therapy were compared. RESULTS: AZA or 6-MP was tolerated in 51 of 95 patients (54%) without adverse reaction; 27 of 95 (28%) experienced side effects that responded to dose reduction (23 patients) or spontaneously (4 patients), most commonly increased aminotransferase level (13.7%). Cessation of therapy was needed in 17 of 95 patients (18%), including recurrent fever (4), pancreatitis (4), gastrointestinal intolerance (4), and recurrent infections (3). Mean prednisone dose decreased from 24.3 to 8.6 mg/day. CONCLUSIONS: AZA and 6-MP were well tolerated in 82% of patients; of these, prednisone reduction occurred in 87%. However, 18% required discontinuation because of hypersensitivity or infectious side effects. PMID- 9753483 TI - Diagnostic accuracy of serological assays in pediatric inflammatory bowel disease. AB - BACKGROUND & AIMS: Accurate serological assays are desirable for the diagnosis of inflammatory bowel disease (IBD) types in the pediatric age group. The aim of this study was to test the diagnostic accuracy of modified assays for perinuclear (p) antineutrophil cytoplasmic antibodies (ANCAs) and anti-Saccharomyces cerevisiae antibodies (ASCAs) in patients with pediatric ulcerative colitis (UC) and Crohn's disease (CD) and in those without IBD. METHODS: With observers blinded to patients' diagnoses, serum specimens were analyzed for immunoglobulin (Ig) A and IgG ASCAs and ANCAs by enzyme-linked immunosorbent assay. The perinuclear location of ANCAs visualized by indirect immunofluorescence was confirmed by its disappearance after administration of deoxyribonuclease. RESULTS: IgA and IgG ASCA titers were significantly greater and highly specific for CD (95% for either, 100% if both positive). pANCA was 92% specific for UC and absent in all non-IBD controls. The majority of patients with CD positive for pANCA had a UC-like presentation. Disease location, duration, activity, complications, and treatment with immunosuppressive drugs did not have an impact on the ASCA or pANCA assay results. After resection, UC patients remained pANCA positive, in contrast to patients with CD, in whom ASCA titers decreased toward normal values postoperatively. CONCLUSIONS: ASCA and pANCA assays are highly disease specific for CD and UC, respectively. These serological tests can assist clinicians in diagnosing and categorizing patients with IBD and may be useful in making therapeutic decisions. PMID- 9753484 TI - The factor V Leiden mutation increases the risk of venous thrombosis in patients with inflammatory bowel disease. AB - BACKGROUND & AIMS: Thromboembolic disease is a significant cause of morbidity and mortality in patients with inflammatory bowel disease (IBD). The aim of this study was to determine the incidence and possible association of the factor V Leiden mutation with the development of thrombosis in patients with IBD. METHODS: This retrospective study included 11 patients with IBD and arterial or venous thrombosis and 51 patients with IBD and no history of thrombosis who were matched for age, sex, ethnic/racial origin, and type of IBD (controls). The presence of the factor V Leiden mutation was determined by coagulation assay and confirmed by a polymerase chain reaction method. RESULTS: Four of 11 IBD patients (36%) with thrombosis and 2 of 51 IBD controls (4%) were heterozygotes for the factor V Leiden mutation (relative risk, 14.00; 95% confidence interval, 1.55-169.25; P = 0.009, Fisher exact test). All thrombotic events in the patients with activated protein C resistance were venous with a calculated prevalence of 50% (4 of 8 patients) and a relative risk of venous thrombosis in IBD patients with factor V Leiden of 23 (95% confidence interval, 2-294; P = 0.005). CONCLUSIONS: In patients with IBD, inheritance of the factor V Leiden mutation results in a significant increased risk of venous thrombosis. PMID- 9753485 TI - Budesonide versus prednisone in the treatment of active Crohn's disease. The Israeli Budesonide Study Group. AB - BACKGROUND & AIMS: Budesonide (BUD) is a potent steroid that undergoes extensive first-pass metabolism. BUD incorporated in a pH-dependent formulation has been proposed as an alternative treatment for Crohn's disease (CD). The aim of this study was to compare the efficacy and safety of BUD and prednisone (PRED) in the treatment of active CD involving the terminal ileum and/or the colon. METHODS: Patients with mild to moderately active CD were included in a randomized, double blind, double-dummy controlled trial. Patients received either 9 mg BUD once daily for 8 weeks or 40 mg PRED once daily for the first 2 weeks tapered gradually to 5 mg/day by the end of the study. Disease activity, quality of life, and laboratory parameters were recorded. RESULTS: One hundred patients received BUD, and 101 patients received PRED. By intention-to-treat analysis, treatment efficacy defined as Crohn's Disease Activity Index of <150 at completion was 51% and 52.5% for the BUD and PRED groups, respectively. Twice as many responded to treatment with no side effects in the BUD compared with the PRED group (30% vs. 14%) (P = 0.006). Most of the decrease in CDAI scores occurred during the first 2 weeks. CONCLUSIONS: BUD is as effective as PRED in the treatment of CD involving the terminal ileum and right colon. BUD has significantly fewer steroid-related adverse reactions. PMID- 9753486 TI - Differential lamina propria cell migration via basement membrane pores of inflammatory bowel disease mucosa. AB - BACKGROUND & AIMS: In active inflammatory bowel disease (IBD), the intestinal mucosa is infiltrated by polymorphonuclear cells (PMNs), lymphocytes, and monocytes from the systemic circulation. Using an ex vivo model, we have investigated luminally directed migration of cells out of the lamina propria. METHODS: Fresh untreated and deepithelialized mucosal samples were studied by electron microscopy. Cells migrating out of the lamina propria were investigated by immunohistochemistry and fluorescence-activated cell sorter analysis. RESULTS: In intact IBD mucosal samples, tunnels containing cells were prominent in the lamina propria matrix, and PMNs, but not other cell types, were prominent in the epithelium. In deepithelialized mucosal samples, the basement membrane was either destroyed or contained numerous large pores. During culture of deepithelialized mucosal samples, many cells (3.3 [+/-0.8] x 10(5) . g tissue-1 . h-1) migrated out of the lamina propria via basement membrane pores. PMNs and eosinophils were prominent during the first 3 hours of culture, but T cells predominated thereafter. Macrophages also migrated, but B cells were the minority population (<2%) at all times. CONCLUSIONS: In active IBD mucosa with an intact epithelium, luminally directed migration of lamina propria cells is restricted mainly to PMNs. After loss of the epithelium, other cell types also migrate into the lumen via numerous, large, basement membrane pores. PMID- 9753487 TI - Consequences of Fas-ligand and perforin expression by colon T cells in a mouse model of inflammatory bowel disease. AB - BACKGROUND & AIMS: We describe a type of colitis that develops after transplantation of nonallogeneic wt bone marrow cells into T cell- and natural killer cell-deficient Tg26 mice (BM-->Tg26). In these animals, severe wasting and inflammation of the colon correlates with the expansion of mucosal T lymphocytes that displays cytotoxic activity. The aims of this study were to determine the relative contribution of perforin and Fas ligand (Fas-L) expression to the cytotoxic action of these T cells and to examine the influence of each pathway in this model of colitis. METHODS: Colonic T cells were tested for their ability to mediate Fas- and perforin-dependent killing in redirected cytotoxicity assays. Bone marrow cells from donor mice lacking either Fas-L (gld mice) or perforin (PFPnull mice) or both molecules were used to reconstitute Tg26 mice. RESULTS: Colon cytotoxic T lymphocyte displayed both Fas- and perforin-dependent killing. Deficiency in perforin, but not Fas-L, resulted in reduced incidence of wasting and, to a lesser extent, severe colitis in BM-->Tg26 animals. CONCLUSIONS: Colon T cells from BM-->Tg26 mice express both perforin and Fas-L. Although neither pathway is critical in the development of colitis, perforin does have a measurable influence on disease in the BM-->Tg26 colitis model. PMID- 9753488 TI - Subtherapeutic corticosteroids potentiate the ability of interleukin 10 to prevent chronic inflammation in rats. AB - BACKGROUND & AIMS: Interleukin (IL)-10, which inhibits macrophages and T-helper lymphocyte type 1 (TH1) lymphocytes, attenuates chronic granulomatous inflammation induced by bacterial cell wall polymers. This study determines whether corticosteroids enhance the protective effects of IL-10 in cultured peripheral blood mononuclear cells (PBMNCs) and in vivo when started before or after the onset of experimental chronic granulomatous inflammation. METHODS: Intestines of Lewis rats were injected intramurally with streptococcal peptidoglycan-polysaccharide (PG-APS) polymers. Daily murine recombinant IL-10 and/or dexamethasone (DEX) therapy was started 12 hours before or at several intervals after PG-APS injection. RESULTS: IL-10 plus corticosteroids additively inhibited IL-1beta secretion in human PBMNCs but preserved the beneficial IL 1RA/IL-1beta ratio induced by IL-10. IL-10 started before PG-APS injection significantly attenuated intestinal and extraintestinal inflammation, with even more pronounced effects in combination with subtherapeutic doses of DEX. The combination of DEX decreased the effective dose of IL-10 by at least one half. After onset of systemic inflammation using doses effective for prevention, IL-10 monotherapy had nearly no benefit and DEX plus IL-10 was similar to the mild therapeutic effect of DEX alone. CONCLUSIONS: The combination of IL-10 and corticosteroids allows lower doses of both agents in preventing chronic intestinal and systemic inflammation. However, timing of IL-10 administration is a critical variable in regulating inflammation. PMID- 9753489 TI - Intraepithelial lymphocytes from villus tip and crypt portions of the murine small intestine show distinct characteristics. AB - BACKGROUND & AIMS: Intraepithelial lymphocytes (IELs) are located between epithelial cells that are thought to display unique features and functions at the small intestinal villus tip and crypt levels. We have addressed whether the spatial differences in the intestinal epithelium extend to IELs and subsequent cross-talk between IELs and epithelial cells. METHODS: IELs were isolated from villus tip and crypt portions of mouse small intestine and then compared for spontaneous cytokine production and responsiveness to interleukin (IL)-2 and/or IL-7. RESULTS: No difference was observed between number of beta IELs in villus tips and crypts, whereas a trend toward increased frequencies of IELs bearing the gamma delta form of T-cell receptor was noted in villus tips. Interestingly, the number of beta IELs producing interferon gamma and IL-5 was significantly reduced in the cells from crypts compared with villus tips. Furthermore, villus tip beta IELs exhibited higher responses to stimulation signals provided by IL-2 and/or IL 7 than their crypt counterpart. Such functional differences were not observed with gamma delta IELs from the two intestinal sites. CONCLUSIONS: Distinct molecular cross-talk between IELs and epithelial cells occurs in intestinal villus tips and crypts. PMID- 9753490 TI - Prostaglandins prevent decreased epithelial cell proliferation associated with dextran sodium sulfate injury in mice. AB - BACKGROUND & AIMS: Although dextran sodium sulfate (DSS)-induced colitis is a commonly used model of colonic injury, the mechanism of this model is not understood. The aim of this study was to determine the contribution of prostaglandins to the mechanism of DSS-induced epithelial injury. METHODS: Mice were treated with 3% DSS in the drinking water for 5 days followed by water only (recovery). Tissue prostaglandin E2 (PGE2) levels were measured, proliferating cells per cecal crypt were determined by bromodeoxyuridine labeling, and the cellular localization of cyclooxygenase (COX)-1 and COX-2 was determined by immunohistochemistry. RESULTS: DSS decreased the number of proliferating epithelial cells per crypt by approximately 90% and decreased the height of cecal crypts by 40%. Administration of dimethyl PGE2 with DSS reversed the effect of DSS on proliferation but not its effect on crypt shortening. COX-1 was expressed in the crypt epithelium and lamina propria mononuclear cells; DSS treatment down regulated COX-1 expression only in the epithelium. Dimethyl PGE2 reversed the effect of DSS on COX-1 expression. Recovery was associated with a return to normal COX-1 expression in the epithelium. COX-2 was expressed in lamina propria mononuclear cells. CONCLUSIONS: Epithelial cell proliferation in the presence of DSS contains a PGE2-sensitive component. PMID- 9753491 TI - Crucial role for 5-HT in cholera toxin but not Escherichia coli heat-labile enterotoxin-intestinal secretion in rats. AB - BACKGROUND & AIMS: Many consider cholera toxin (CT) and Escherichia coli heat labile enterotoxin (LT) to be functionally identical. Both increase intracellular adenosine 3',5'-cyclic monophosphate concentration; however, differences between the two and the severity of the diseases they cause have been reported. The secretagogue 5-hydroxytryptamine (5-HT) is implicated in CT-induced secretion, but its role in LT-induced secretion is unclear. We tested the hypothesis that LT fails to recruit 5-HT in its secretory processes. METHODS: In vivo small intestinal perfusions were undertaken in adult male Wistar rats after incubation with equipotent doses of CT or LT, or saline. Small intestinal 5-HT release and the effect on net small intestinal water and electrolyte transport of (1) pharmacological depletion of 5-HT; (2) blockade of 5-HT type 2, 3, and 4 receptors; and (3) pretreatment with lidocaine, hexamethonium, and atropine were determined. RESULTS: CT- but not LT-induced secretion was accompanied by 5-HT release, reduced by 5-HT depletion, and inhibited by each 5-HT antagonist. By contrast, lidocaine and hexamethonium inhibited secretion induced by both toxins. CONCLUSIONS: LT induces secretion without recruiting a 5-HT-dependent cascade. This may account for differences in clinical severity of the diseases CT and LT cause and has implications for the development of antisecretory therapies. PMID- 9753492 TI - Immunization of BALB/c mice with Helicobacter urease B induces a T helper 2 response absent in Helicobacter infection. AB - BACKGROUND & AIMS: Infection with Helicobacter induces a T helper type 1 response in mice and humans. Mice can be cured or protected from infection with Helicobacter by mucosal immunization with recombinant H. pylori urease B subunit (rUreB). This study characterizes the immune response of infected mice immunized with rUreB. METHODS: BALB/c mice were infected with H. felis. Two weeks later, they were orally immunized four times with rUreB and cholera toxin (CT) at weekly intervals. Controls were only infected or sham-immunized with CT. Animals were killed at various times after immunization. Splenic CD4(+) cells were obtained and cultured in vitro with rUreB to evaluate antigen-specific proliferation and induction of interferon gamma and interleukin 4 secretion. RESULTS: All rUreB immunized mice (n = 8) were cured from infection 3 weeks after the fourth immunization. Immunization induced a proliferative response of splenic CD4(+) cells, a progressive decrease in interferon gamma secretion, and a concomitant increase in interleukin 4 secretion after each immunization. A simultaneous increase in rUreB specific serum immunoglobulin G1 levels was observed in infected/immunized mice. CONCLUSIONS: In BALB/c mice, therapeutic mucosal immunization with rUreB induces progressively a Th2 CD4(+) T cell response resulting in the elimination of the pathogen. PMID- 9753493 TI - Identification of neurons that express stem cell factor in the mouse small intestine. AB - BACKGROUND & AIMS: Enteric neurons in the murine intestine express stem cell factor (SCF), which may provide an important signal in the development of the interstitial cells of Cajal (ICC). Our aim was to identify the subpopulation(s) of myenteric neurons that express SCF. METHODS: Myenteric plexus preparations from postnatal SCF-lacZ mice were processed for beta-galactosidase histochemistry followed by immunohistochemistry. RESULTS: Approximately 60% of the nitric oxide synthase-immunoreactive neurons, which projected to myenteric ganglia and to circular muscle, expressed SCF, and more than 80% of the calbindin-immunoreactive neurons, which projected exclusively to myenteric ganglia, expressed SCF. A small subpopulation of calretinin-immunoreactive neurons expressed SCF transiently. Many of the remainder of SCF-expressing neurons were choline acetyltransferase immunoreactive, but their projections are unknown. CONCLUSIONS: SCF-expressing neurons that project within the myenteric plexus may be an important source of SCF for the development of Kit-expressing ICC at this level. The only possible neuronal source of SCF for the ICC of the deep muscular plexus is a subpopulation of nitric oxide synthase-immunoreactive neurons. PMID- 9753494 TI - Electrophysiology, shape, and chemistry of neurons that project from guinea pig colon to inferior mesenteric ganglia. AB - BACKGROUND & AIMS: Prevertebral sympathetic ganglia receive inputs from intestinofugal neurons, with cell bodies located in the wall of the bowel. Intestinofugal neurons are part of the afferent limbs of intestino-intestinal reflexes. The aim of this study was to define the properties of intestinofugal neurons using intracellular recordings. METHODS: Intestinofugal neurons of the distal colon were retrogradely labeled from the inferior mesenteric ganglia. In whole mounts of the myenteric plexus/longitudinal muscle of the distal colon, labeled neurons were identified by their fluorescence and recordings were made using biocytin-filled electrodes. Labeled nerves were characterized immunohistochemically and morphologically. RESULTS: Intestinofugal neurons were uniaxonal neurons with multiple dendrites that had lamellar expansions. They were immunoreactive for choline acetyltransferase. Stimulation of nerve fiber tracts elicited large-amplitude excitatory postsynaptic potentials in all labeled neurons. Some received spontaneous fast excitatory postsynaptic potentials. Those cells that fired action potentials fired only one or two at the start of a depolarizing current pulse. No intestinofugal neurons had Dogiel type II morphology or a late afterhyperpolarizing potential. CONCLUSIONS: Intestinofugal neurons are likely to be activated by other neurons in the gut wall. They are not AH or Dogiel type II neurons. Thus they seem to be second order neurons in afferent pathways of intestino-intestinal reflexes. PMID- 9753495 TI - Leukotriene D4-induced contraction of cat esophageal and lower esophageal sphincter circular smooth muscle. AB - BACKGROUND & AIMS: In esophageal circular muscle, acetylcholine activates phosphatidylcholine-specific phospholipases C and D and phospholipase A2, producing diacylglycerol and arachidonic acid, which cause contraction by interacting synergistically to activate protein kinase C. In a model of acute esophagitis, leukotriene D4 (LTD4) contributes to acetylcholine-induced contraction. We examined intracellular signaling in LTD4-induced contraction. METHODS: Esophageal and lower esophageal sphincter (LES) cells, isolated by enzymatic digestion, were contracted by LTD4 in the absence or presence of inhibitors. Permeabilization by saponin allowed use of G-protein antibodies and heparin. RESULTS: Esophageal contraction was inhibited by pertussis toxin, Gi3 antibodies, D609 (phosphatidylcholine-specific phospholipase C inhibitor), propranolol (phospholipase D pathway inhibitor), and chelerythrine (protein kinase C antagonist) but not W7 (calmodulin antagonist). LES contraction was unaffected by pertussis toxin. It was inhibited by Gq antibodies, U-73122 (phosphatidylinositol-specific phospholipase C inhibitor), heparin (inositol 1,4,5-trisphosphate inhibitor), and W7 and reduced by D609. CONCLUSIONS: In the esophagus, LTD4 activates a protein kinase C-dependent pathway through pertussis toxin-sensitive Gi3 proteins and phosphatidylcholine-specific phospholipase. In the LES, LTD4 activates a calmodulin-dependent pathway through pertussis toxin insensitive Gq proteins and phosphatidylinositol-specific phospholipase C. The intracellular pathways activated by LTD4 in the esophagus and the LES are similar to those activated by acetylcholine and other agonists. PMID- 9753496 TI - Noninvasive prediction of fibrosis in C282Y homozygous hemochromatosis. AB - BACKGROUND & AIMS: The diagnosis of hemochromatosis is now possible for C282Y homozygous patients using noninvasive molecular genetic tests. The aim of this study was to define noninvasive factors predictive of severe fibrosis (bridging fibrosis or cirrhosis) to avoid unnecessary liver biopsies in such patients. METHODS: Clinical and biological data were recorded at the time of diagnosis in 197 French C282Y homozygous patients, 52 (26%) of whom had severe fibrosis. Variables significantly linked to severe fibrosis using univariate analysis were entered into a multivariate stepwise analysis. These variables were combined to obtain a simple index allowing for prediction of severe fibrosis. RESULTS: Serum ferritin, hepatomegaly, and serum aspartate aminotransferase were selected using multivariate analysis. Their combination applied to the 96 patients with ferritin level of 0.05). CONCLUSIONS: Asthma may lead to physiological features similar to COPD but may be distinguished by demonstrating a preserved DLCO and a higher ratio of airway to parenchymal abnormalities on HRCT scan. PMID- 9753529 TI - Canadian Thoracic Society guidelines for occupational asthma. AB - OBJECTIVE: To provide broad guidelines and principles to help primary care physicians, occupational physicians, allergists and respirologists with the recognition, diagnosis and management of patients with occupational asthma (OA). OPTIONS: These guidelines are mainly directed towards OA induced by a workplace sensitizing agent. However, irritant-induced asthma and workplace aggravation of underlying asthma are also addressed, and some consideration is given to other differential diagnoses. OUTCOMES: To enable the assessing physician to investigate patients with possible OA appropriately and to provide guidelines for appropriate early referral when specialized investigations are required. To provide an understanding of the appropriate management strategies following objective diagnosis. EVIDENCE: The key diagnostic and management recommendations were based on a critical review of the literature and by specialist consensus meetings. VALUES: Evidence was categorized as follows. Level 1: Evidence from at least one randomized, controlled trial. Level 2: Evidence from at least one well designed clinical trial without randomization, from cohort or case-control analytical studies, preferably from more than one centre, from multiple time series or from dramatic results in uncontrolled experiments. Level 3: Evidence from the opinions of respected authorities based on clinical experience, descriptive studies or reports of expert committees. Evidence was further subdivided as follows: A. Good evidence to support a recommendation for use; B. Moderate evidence to support a recommendation for use; C. Poor evidence to support a recommendation for or against use; D. Moderate evidence to support a recommendation against use; E. Good evidence to support a recommendation against use. BENEFITS, HARM AND COSTS: The medical and socioeconomic risks and benefits of an incorrect diagnosis of OA and of failure to diagnose true OA were considered in the recommendations. VALIDATION: The document has been reviewed and endorsed by the Canadian Thoracic Society, the Canadian Society of Allergy and Clinical Immunology, and The College of Family Physicians of Canada. CONCLUSIONS: There is good evidence for rapid investigation and objective categorization of presented symptoms into OA, aggravation of underlying asthma, unrelated asthma or other diagnoses. OA should be suspected in all adult onset asthmatics whose asthma begins or worsens while they are working. Investigations should be directed to an objective assessment of asthma and then to an assessment of the work relationship, using a combination of investigations as feasible, which may include immunological tests, pulmonary function assessed during work periods and away from work, and specific challenge tests. Early specialist referral is recommended for diagnosis. Management strategies include general asthma management in addition to measures to avoid further exposure to a relevant workplace sensitizer. Compensation issues and other workers at risk of developing OA also need to be considered when the diagnosis is made. PMID- 9753530 TI - Mycotic pulmonary arterial aneurysms in an intravenous drug user. AB - A case of mycotic pulmonary artery aneurysm (PAA) in an intravenous drug user in whom resolution occurred with conservative therapy is described. The natural history of PAA is not well described in the literature. Although PAA is potentially fatal, resolution may occur in patients who do not have hemoptysis. Clinical presentation, diagnosis and management of PAA are reviewed. PMID- 9753531 TI - Varieties of deficit unawareness after brain injury. AB - The failure to recognize the existence of disease is known as anosognosia. This article provides a brief discussion of the evolution of this concept and reviews some qualitative differences in the manner in which the disavowal of neurogenic impairment is expressed. Theoretical explanations for the unawareness of deficit after neurologic illness include motivational-symbolic, cognitive subsystem, and supraordinate system theories. Observations from a clinically derived, structured awareness interview are presented, suggesting three factors that may underlie patients' apparent lack of awareness of deficits after traumatic brain injury. These include diminished awareness of deficits secondary to impaired cognition, especially memory and reasoning deficits; psychological reactance and denial of deficits; and a relatively "pure" inability to recognize areas of impaired functioning as a direct consequence of brain injury. The causes of unawareness are likely to be complex and multiply determined in any given patient, although it may be possible to identify primary, secondary, and even tertiary contributions according to specific behavioral and phenomenological characteristics. The ability of patients to modify their perceptions and acknowledgment of deficits after objective feedback may have particular diagnostic value and clinical utility in this regard. PMID- 9753532 TI - Awareness of errors in naturalistic action after traumatic brain injury. AB - A prospective study was performed to develop a method for assessing "on-line" error detection and correction during performance of naturalistic action, to determine whether traumatic brain injury (TBI) affects error detection and correction, and to compare actual task performance with verbal self-ratings of performance. Participants included 18 persons who had sustained severe TBI from 34 to 186 days prior to study and who were comparable to controls in their rate of naturalistic action error, along with 18 control subjects chosen to be demographically comparable to subjects with TBI. Subjects performed two different tests of naturalistic action in which they completed everyday activities (eg, wrapping a gift, making toast) at different levels of complexity, as manipulated by the addition of distractor objects, the number of tasks that had to be completed per trial, and other demands on planning and working memory. Using a specially developed coding system, each error on these tasks was scored as to whether the subject corrected it and whether the subject otherwise demonstrated awareness of the error. Error scores were also compared to subjects' responses to a questionnaire in which they rated their own performance on the most challenging level of the naturalistic action test. In general, subjects with TBI corrected and showed awareness of proportionally fewer of their errors when compared to controls. Qualitative patterns for some error types also differed between groups. Despite making more errors than control subjects on the most challenging task, subjects with TBI did not rate themselves as performing more poorly with respect to its cognitive demands. However, for subjects with TBI, the number of errors was correlated with performance ratings on certain questionnaire items. This study showed that error detection and correction can be reliably measured during naturalistic action and appear to be impaired in severe TBI even when the base rate of error is controlled. TBI may affect error detection and correction by reducing, or impairing the allocation of, attentional resources needed for the simultaneous execution and monitoring of routine action. PMID- 9753533 TI - Initial disturbances of consciousness and resultant impaired awareness in Spanish patients with traumatic brain injury. AB - The purpose of this prospective, between-subjects study was to look at impaired awareness cross-culturally in patients with traumatic brain injury (TBI) and to relate impaired awareness after injury to the initial estimates of disturbed consciousness at time of injury. The study was conducted in community and inpatient and outpatient rehabilitation centers in Barcelona and Madrid. Participants were 30 persons with primarily moderate to severe TBI who could complete a written questionnaire concerning their functioning and 28 age- and gender-matched controls. A Spanish translation of the Patient Competency Rating Scale (PCRS) was administered to each participant. Relatives or significant others also completed this scale on each participant using the relative's version (PCRS-R). Difference scores, obtained by subtracting PCRS-R from PCRS-P (PCRS-P minus PCRS-R), were used as a marker of impaired awareness. Individuals with TBI were rated (by self and significant others) as being less competent than controls. Forty percent of Spanish patients with TBI who suffered severe injuries tended to overestimate their behavioral competencies. The PCRS-P minus the PCRS-R difference scores tended to correlate with admitting Glasgow Coma Scale (GCS) scores and retrospective estimates of posttraumatic amnesia (PTA). Initial disturbances of consciousness, one measure of severity of brain injury, appeared to relate to later measures of impaired self-awareness in Spanish patients with TBI. Non-brain-injured controls did not tend to report levels of competency that differed from their relatives' reports. PMID- 9753534 TI - Cluster analysis of self-awareness levels in adults with traumatic brain injury and relationshipto outcome. AB - The purpose of this study was to investigate the relationship between self awareness, emotional distress, motivation, and outcome in adults with severe traumatic brain injury. A sample of 55 patients were selected from 120 consecutive patients with severe traumatic brain injury admitted to the rehabilitation unit of a large metropolitan public hospital. Subjects received multidisciplinary inpatient rehabilitation and different types of outpatient rehabilitation and community-based services according to availability and need. Measures used in the cluster analysis were the Patient Competency Rating Scale, Self-Awareness of Deficits Interview, Head Injury Behavior Scale, Change Assessment Questionnaire, the Beck Depression Inventory, and Beck Anxiety Inventory; outcome measures were the Disability Rating Scale, Community Integration Questionnaire, and Sickness Impact Profile. A three-cluster solution was selected, with groups labeled as high self-awareness (n = 23), low self awareness (n = 23), and good recovery (n = 8). The high self-awareness cluster had significantly higher levels of self-awareness, motivation, and emotional distress than the low self-awareness cluster but did not differ significantly in outcome. Self-awareness after brain injury is associated with greater motivation to change behavior and higher levels of depression and anxiety; however, it was not clear that this heightened motivation actually led to any improvement in outcome. Rehabilitation timing and approach may need to be tailored to match the individual's level of self-awareness, motivation, and emotional distress. PMID- 9753536 TI - Awareness intervention: who needs it? AB - It is widely accepted that awareness deficits present challenges to recovery and should be addressed as part of rehabilitation programming. Response to awareness intervention is commonly inferred from measurements that rely on reports by subjects and significant others. This article describes the findings from a pilot study that examined the relationship among a variety of awareness indicators in three individuals with brain damage over a 9-month period. Results suggest a dissociation between behavioral and perceptual indices of awareness. Changes in behavioral indicators of awareness selected by caregivers were not related to changes in self- or caregiver ratings. The clinical and research implications of the findings are discussed. PMID- 9753535 TI - Impaired awareness and employment outcome after traumatic brain injury. AB - Inaccurate self-awareness is a common finding after traumatic brain injury. Such impaired awareness has been hypothesized to limit patients' eventual functional outcomes by decreasing motivation for treatment and resulting in selection of inappropriate long-term goals. Previous investigations of the association between impaired awareness and employment outcome have produced inconsistent results. The present article reviews these studies and presents the results of our new investigation of this issue. In addition, we studied the comparability of two methods of measuring impaired awareness. Results provided strong support for a positive relationship between accurate self-awareness and favorable employment outcome at follow-up. PMID- 9753537 TI - Sociological and cultural aspects in postacute neuropsychological rehabilitation. AB - Various forms of postacute brain injury rehabilitation programs exist. Comprehensive day treatment programs are the most holistic, being neuropsychological in orientation. The importance of developing a trusting relationship and the use of inventive techniques in the individualization of treatment within a holistic rehabilitation program will be described via a case presentation, stressing the sociocultural aspects of the individual as central to treatment. PMID- 9753538 TI - Methylphenidate for cognitive and behavioral dysfunction after traumatic brain injury. PMID- 9753539 TI - Software update 1998: commercial programs useful in cognitive retraining. PMID- 9753540 TI - Communication and ethics: cardiopulmonary resuscitation in head trauma rehabilitation. PMID- 9753541 TI - Brain injury advocacy. PMID- 9753542 TI - Preface PMID- 9753543 TI - Phosphorylation stabilizes the active conformation of rhodopsin PMID- 9753544 TI - Cofilin and gelsolin segment-1: molecular dynamics simulation and biochemical analysis predict a similar actin binding mode. AB - An understanding of the actin-depolymerizing function attributed to members of the ADF/cofilin/destrin superfamily requires a structural model of these proteins in complex with actin. As a step toward defining actin-cofilin interactions, the complex of yeast cofilin with monomeric actin was predicted, starting with the actin-gelsolin segment-1 binding mode recently suggested for the actin-destrin complex. After refinement by molecular dynamics simulation, the structure of cofilin converged in a new binding mode that required only minimal changes induced in the actin-cofilin interface. The predicted complex exhibits strong interactions between the N termini of actin and cofilin, mediated by a salt bridge of cofilin Arg3 with actin Asp1. The forming of this salt bridge could be prevented by the phosphorylation of cofilin Ser4, which is believed to inhibit cofilin depolymerization activity. Recent mutagenesis studies, crosslinking experiments and peptide binding studies are consistent with the predicted model of the actin-cofilin complex. The structural homology between cofilin and gelsolin segment-1 binding to actin was confirmed experimentally by two types of competitive binding assays. PMID- 9753546 TI - Cooperative non-specific DNA binding by octamerizing lambda cI repressors: a site specific thermodynamic analysis. AB - Relationships between dimerization and site-specific binding have been characterized previously for wild-type and mutant cI repressors at the right operator (OR) of bacteriophage lambda DNA. However, the roles of higher-order oligomers (tetramers and octamers) that are also formed from these cI molecules have remained elusive. In this study, a clear correlation has been established between repressor oligomerization and non-specific DNA-binding activity. A modification of the quantitative DNase I footprint titration technique has been used to evaluate the degree of saturation of non-specific, OR-flanking lambda DNA by cI repressor oligomers. With the exception of one mutant, only those repressors capable of octamerizing were found to exhibit non-specific DNA-binding activity. The non-specific interaction was accurately modeled using either a one dimensional, univalent, site-specific Ising lattice approximation, or a more traditional, multivalent lattice approach. It was found that non-specific DNA binding by repressor oligomers is highly cooperative and energetically independent from site-specific binding at OR. Furthermore, the coupling free energy resolved for non-specific binding was similar to that of site-specific binding for each repressor, suggesting that similar structural elements may mediate the cooperative component of both binding processes. It is proposed that the state of assembly of the repressor molecule modulates its relative affinity for specific and non-specific DNA sequences. These specificities are allosterically regulated by the transmission of assembly-state information from the C-terminal domain, which mediates self-association and cooperativity, to the N-terminal domain, which primarily mediates DNA-binding. While dimers have a high affinity for their cognate sites within OR, tetramers and octamers may preferentially recognize non-specific DNA sequences. The concepts and findings developed in this study may facilitate quantitative characterization of the relationships between specific, and non-specific binding in other systems that utilize multiple modes of DNA-binding cooperativity. PMID- 9753545 TI - TOM1p, a yeast hect-domain protein which mediates transcriptional regulation through the ADA/SAGA coactivator complexes. AB - The hect-domain has been characterized as a conserved feature of a group of E3 ubiquitin ligases. Here we show that the yeast hect-domain protein TOM1p regulates transcriptional activation through effects on the ADA transcriptional coactivator proteins. Null mutations of tom1 result in similar defects in transcription from ADH2 and HIS3 promoters, and enhanced transcription from the GAL10 promoter as do null mutations in ngg1/ada3. Strains with disruptions of both ngg1 and tom1 have the same phenotype as strains with a disruption of only ngg1 implying that these genes are acting through the same pathway. In the absence of TOM1p, the normal associations of the ADA proteins with SPT3p and the TATA-binding protein are reduced. The action of TOM1p is most likely mediated through ubiquitination since mutation of Cys3235 to Ala, corresponding residues of which are required for thioester bond formation with ubiquitin in other hect domain proteins, results in similar changes in transcription as the null mutation. A direct role for TOM1p in regulation of ADA-associated proteins is further supported by the finding that SPT7p is ubiquitinated in a TOM1p-dependent fashion and that TOM1p coimmunoprecipitates with the ADA proteins. PMID- 9753547 TI - Transmembrane helix tilting and ligand-induced conformational changes in the lactose permease determined by site-directed chemical crosslinking in situ. AB - The N-terminal six transmenbrane helices (N6) and the C-terminal six transmembrane helices (C6) of the lactose permease of Escherichia coli, each with a Cys residue, were co-expressed independently, and crosslinking was studied. Proximity of paired Cys residues in helices II (position 49, 52, 53, 56, 57, 60, 63 or 67) and VII (position 227, 230, 231, 234, 238, 241, 242 or 245) or XI (position 350, 353, 354, 357, 361 or 364) was examined by using two homobifunctional thiol-specific crosslinking agents of different lengths (6 or 10 A). The results demonstrate that a Cys residue placed in the periplasmic half of helix II (position 49, 52, 53 or 57) crosslinks to Cys residues in the periplasmic half of helix VII (position 241, 242 or 245). In contrast, no crosslinking is evident with paired-Cys residues in the cytoplasmic halves of helices II (position 60, 63 or 67) and VII (position 227, 230, 231, 234 or 238). Remarkably, a Cys residue in the cytoplasmic half of helix II (position 60, 63 or 67) crosslinks with a Cys residue in the cytoplasmic half of helix XI (position 350, 353 or 354), while paired-Cys residues at positions in the periplasmic halves of the two helices do not crosslink. Therefore, helix II is tilted in such a manner that the periplasmic end is close to helix VII, and the cytoplasmic end is close to helix XI. Furthermore, ligand-binding alters the crosslinking efficiency of paired-Cys residues in helices II and VII or XI, indicating that both interfaces are conformationally active. The results are consistent with the conclusion that ligand-binding induces a scissors-like movement of helices II and VII that increases interhelical distance by 3 to 4 A at the periplasmic ends and decreases the distance by 3 to 4 A at the approximate middle of the two transmembrane helices. PMID- 9753549 TI - Characterisation of an improved two-dimensional p22121 crystal from bovine rhodopsin. AB - Dialysis of rhodopsin isolated from bovine rod outer segments resulted in the formation of a new two-dimensional crystal form suitable for electron crystallography. The crystals obtained were tubular or single layers and showed p22121 symmetry (a=60.6(+/-0.8) A, b=86.3(+/-1.6) A). For the first time the size and order of the crystals allowed us to take electron diffraction patterns showing spots to a resolution of about 3.5 A. Images were recorded at liquid nitrogen temperature using a high voltage field emission electron microscope. Out of a large number of images 20 crystalline areas were selected and processed with the MRC image processing software. A projection structure of bovine rhodopsin to 5 A resolution was calculated using amplitudes and phases extracted from these images. The achieved resolution exceeds the resolution of all previously obtained structures of frog, bovine and squid rhodopsin crystals. In this map small differences are observed compared to the previous maps. Helix 5 seems to be even more highly tilted and between the arc-shaped feature and helix 5 a peak is present suggesting that helix 3 is prolonging this feature towards helix 5. These observations are in agreement with the latest model for the three-dimensional arrangement of rhodopsin. The resolution achieved as well as the availability of electron diffraction data suggest that there is a good possibility to collect data from tilted crystals and calculate an improved three-dimensional structure of rhodopsin. PMID- 9753548 TI - Complete sequence of the IncPbeta plasmid R751: implications for evolution and organisation of the IncP backbone. AB - The broad host range IncP plasmids are of particular interest because of their ability to promote gene spread between diverse bacterial species. To facilitate study of these plasmids we have compiled the complete sequence of the IncPbeta plasmid R751. Comparison with the sequence of the IncPalpha plasmids confirms the conservation of the IncP backbone of replication, conjugative transfer and stable inheritance functions between the two branches of this family. As in the IncPalpha genome the DNA of this backbone appears to have been enriched for the GCCG/CGGC motifs characteristic of the genome of organisms with a high G+C content, such as P. aeruginosa, suggesting that IncPbeta plasmids have been subjected during their evolution to similar mutational and selective forces as IncPalpha plasmids and may have evolved in pseudomonad hosts. The IncP genome is consistently interrupted by insertion of phenotypic markers and/or transposable elements between oriV and trfA and between the tra and trb operons. The R751 genome reveals a family of repeated sequences in these regions which may form the basis of a hot spot for insertion of foreign DNA. Sequence analysis of the cryptic transposon Tn4321 revealed that it is not a member of the Tn21 family as we had proposed previously from an inspection of its ends. Rather it is a composite transposon defined by inverted repeats of a 1347 bp IS element belonging to a recently discovered family which is distributed throughout the prokaryotes. The central unique region of Tn4321 encodes two predicted proteins, one of which is a regulatory protein while the other is presumably responsible for an as yet unidentified phenotype. The most striking feature of the IncPalpha plasmids, the global regulation of replication and transfer by the KorA and KorB proteins encoded in the central control operon, is conserved between the two plasmids although there appear to be significant differences in the specificity of repressor-operator interactions. The importance of these global regulatory circuits is emphasised by the observation that the operator sequences for KorB are highly conserved even in contexts where the surrounding region, either a protein coding or intergenic sequence, has diverged considerably. There appears to be no equivalent of the parABCDE region which in the IncPalpha plasmids provides multimer resolution, lethality to plasmid-free segregants and active partitioning functions. However, we found that the continuous sector from co ordinate 0 to 9100 bp, encoding the co-regulated klc and kle operons as well as the central control region, could confer a high degree of segregational stability on a low copy number test vector. Thus R751 appears to exhibit very clearly what was first revealed by study of the IncPalpha plasmids, namely a fully functional co-ordinately regulated set of replication, transfer and stable inheritance functions. PMID- 9753550 TI - A thymine-like base analogue forms wobble pairs with adenine in a Z-DNA duplex. AB - The DNA hexamer d(CACGPG), in which dP is the ambivalent pyrimidine nucleoside analogue 2'-deoxy-beta-d-ribofuranosyl-(6H,8H-3, 4-dihydropyrimido[4,5 c][1,2]oxazin-7-one), crystallises as a left-handed Z-DNA duplex. X-ray analysis at 1.5 A shows that both P. A base-pairs are of the wobble type. This result appears inconsistent with other evidence from hybridisation and NMR studies of P containing oligonucleotides, which suggests that, while P can form stable base pairs with either A or G, thymine-like properties are more pronounced. Thermal denaturation experiments over a range of solution pH values indicate that protonation of the P.A base-pairs is unlikely to be responsible for the anomalous behaviour. No specific crystal packing effects can be identfied as an explanation, and it is concluded that base stacking and other interactions between nucleotide residues in Z-DNA are responsible. PMID- 9753551 TI - Solution-state structure of a DNA dodecamer duplex containing a Cis-syn thymine cyclobutane dimer, the major UV photoproduct of DNA. AB - The solution structures of a duplex DNA dodecamer containing a cis-syn cyclobutane thymine dimer d(GCACGAAT[cs]TAAG).d(CTTAATTCG TGC) and its native parent sequence were determined using NMR data collected at 750 MHz. The dodecamer sequence corresponds to the section of a site-specific cis-syn dimer containing 49-mer that was found to be the binding site for the dimer-specific T4 denV endonuclease V repair enzyme by chemical and enzymatic footprinting experiments. Structures of both sequences were derived from NOE restrained molecular dynamics/simulated annealing calculations using a fixed outer layer of water and an inner dynamic layer of water with sodium counterions. The resulting structures reveal a subtle distortion to the phosphodiester backbone in the dimer containing sequence which includes a BII phosphate at the T9pA10 junction immediately 3' to the dimer. The BII phosphate, established experimentally by analysis of the 31P chemical shifts and interpretation of the 3JP-H3' values using an optimized Karplus relationship, enables the DNA helix to accommodate the dimer by destacking the base 3' to the dimer. Furthermore, the structures provide explanations for the unusually shifted T8-N3H imino, A16-H2 and T8-Me proton resonances and T9pA10 (31)P NMR resonance and are consistent with bending, unwinding, and thermodynamic data. The implications of the structural data for the mechanism by which cis-syn dimers are recognized by repair enzymes and bypassed by DNA polymerases are also discussed. PMID- 9753552 TI - A comparison of the crystallographic structures of two catalytic antibodies with esterase activity. AB - The crystallographic structure of the Fab fragment of the catalytic antibody, 29G11, complexed with an (S)-norleucine phenyl phosphonate transition state analog was determined at 2.2 A resolution. The antibody catalyzes the hydrolysis of norleucine phenyl ester with (S)-enantioselectivity. The shape and charge complementarity of the binding pocket for the hapten account for the preferential binding of the (S)-enantiomer of the substrate. The structure is compared to that of the more catalytically efficient antibody, 17E8, induced by the same hapten transition state analog. 29G11 has different residues from 17E8 at eight positions in the heavy chain, including four substitutions in the hapten-binding pocket: A33V, S95G, S99R and Y100AN, and four substitutions at positions remote from the catalytic site, I28T, R40K, V65G and F91L. The two antibodies show large differences in the orientations of their variable and constant domains, reflected by a 32 degrees difference in their elbow angles. The VL and VH domains in the two antibodies differ by a rotation of 8.8 degrees. The hapten binds in similar orientations and locations in 29G11 and 17E8, which appear to have catalytic groups in common, though the changes in the association of the variable domains affect the precise positioning of residues in the hapten-binding pocket. PMID- 9753553 TI - The 1.25 A resolution refinement of the cholera toxin B-pentamer: evidence of peptide backbone strain at the receptor-binding site. AB - Crystals of the 61 kDa complex of the cholera toxin B-pentamer with the ganglioside GM1 receptor pentasaccharide diffract to near-atomic resolution. We have refined the crystallographic model for this complex using anisotropic displacement parameters for all atoms to a conventional crystallographic residual R=0.129 for all observed Bragg reflections in the resolution range 22 A to 1.25 A. Remarkably few residues show evidence of discrete conformational disorder. A notable exception is a minority conformation found for the Cys9 side-chain, which implies that the Cys9-Cys86 disulfide linkage is incompletely formed. In all five crystallographically independent instances, the peptide backbone in the region of the receptor-binding site shows evidence of strain, including unusual bond lengths and angles, and a highly non-planar (omega=153.7(7) degrees) peptide group between residues Gln49 and Val50. The location of well-ordered water molecules at the protein surface is notable reproduced among the five crystallographically independent copies of the peptide chain, both at the receptor-binding site and elsewhere. The 5-fold non-crystallographic symmetry of this complex allows an evaluation of the accuracy, reproducibility, and derived error estimates from refinement of large structures at near-atomic resolution. We find that blocked-matrix treatment of parameter covariance underestimates the uncertainty of atomic positions in the final model by approximately 10% relative to estimates based either on full-matrix inversion or on the 5-fold non crystallographic symmetry. PMID- 9753555 TI - Folding and association of beta-Galactosidase. AB - beta-D-Galactosidase from Escherichia coli is one of the largest tetrameric enzymes known at present. Although its physiological importance, the regulation of its synthesis, its enzymatic properties and its structure are well established, little is known about the stability and the folding pathway of this enzyme. Here we show that the overall folding mechanism of chemically denatured beta-galactosidase consists of three stages: (i) formation of elements of secondary structure; (ii) collapse to subdomains and structured monomers; (iii) association to the native quaternary structure via dimeric intermediates. The first rate-limiting step is the association of structured monomers to form dimers in a bi-molecular reaction, with a rate constant of 4.3x10(3) M-1 s-1 at 20 degreesC. The second rate-limiting uni-molecular folding step leads to dimers which are competent for further association, with a rate constant of 0.5x10(-3) s 1 at 20 degreesC. Tetramers form from these dimers in a fast reaction. By determining a similar mechanism for alpha-complementation of beta-galactosidase fragments it could be confirmed that beta-galactosidase follows a consecutive bi uni-molecular mechanism of folding and association. PMID- 9753556 TI - Uncertainty, Sensitivity, Convergence, and Rounding in Performing and Reporting Least-Squares Fits. AB - This paper describes a procedure for optimal rounding of parameters determined from a linear or nonlinear least-squres fit in order to minimize the number of digits which must be quoted while ensuring that the resulting rounded constants can predict the input data with no significant loss of precision. Related problems concerning nonlinear least-squares convergence and taking account of model dependence of fitted or predicted parameters are also addressed. The recommended rounding procedure is illustrated by applications to electronic band data for the A-X system of I2 and to infrared and microwave data for HF (yielding optimal new Dunham expansion coefficients for ground state HF). An automated version of this sequential rounding procedure has been incorporated in a general subroutine for performing linear or nonlinear least-squares fits. Copyright 1998 Academic Press. PMID- 9753554 TI - Structures of native and complexed complement factor D: implications of the atypical His57 conformation and self-inhibitory loop in the regulation of specific serine protease activity. AB - Factor D is a serine protease essential for the activation of the alternative pathway of complement. The structures of native factor D and a complex formed with isatoic anhydride inhibitor were determined at resolution of 2.3 and 1.5 A, respectively, in an isomorphous monoclinic crystal form containing one molecule per asymmetric unit. The native structure was compared with structures determined previously in a triclinic cell containing two molecules with different active site conformations. The current structure shows greater similarity with molecule B in the triclinic cell, suggesting that this may be the dominant factor D conformation in solution. The major conformational differences with molecule A in the triclinic cell are located in four regions, three of which are close to the active site and include some of the residues shown to be critical for factor D catalytic activity. The conformational flexibility associated with these regions is proposed to provide a structural basis for the previously proposed substrate induced reversible conformational changes in factor D. The high-resolution structure of the factor D/isatoic anhydride complex reveals the binding mode of the mechanism-based inhibitor. The higher specificity towards factor D over trypsin and thrombin is based on hydrophobic interactions between the inhibitor benzyl ring and the aliphatic side-chain of Arg218 that is salt bridged with Asp189 at the bottom of the primary specificity (S1) pocket. Comparison of factor D structural variants with other serine protease structures revealed the presence of a unique "self-inhibitory loop". This loop (214-218) dictates the resting state conformation of factor D by (1) preventing His57 from adopting active tautomer conformation, (2) preventing the P1 to P3 residues of the substrate from forming anti-parallel beta-sheets with the non-specific substrate binding loop, and (3) blocking the accessibility of Asp189 to the positive1y charged P1 residue of the substrate. The conformational switch from resting-state to active-state can only be induced by the single macromolecular substrate, C3b-bound factor B. This self-inhibitory mechanism is highly correlated with the unique functional properties of factor D, which include high specificity toward factor B, low esterolytic activity toward synthetic substrates, and absence of regulation by zymogen and serpin-like or other natural inhibitors in blood. PMID- 9753557 TI - Ab Initio Determination of the Torsion-Wagging and Wagging-Bending Infrared Band Structure Spectrum of Methylamine. AB - The infrared band structure for the methyl torsion and amine hydrogen symmetric wagging in methylamine is calculated by ab initio procedures. The influence of the amine hydrogen symmetric bending on the wagging spectrum is considered explicitly. For this purpose, the potential energy surfaces and kinetic parameters were determined at the RHF/MP2 level with the 6-311G++(3df, 3dp) basis set. The numerical results were fitted to symmetry adapted functional forms. The Schrodinger equations for the nuclear motions were solved by expanding the solutions into products of trigonometric functions. The band frequencies and intensities were calculated from the energy levels, the vibrational functions, and the electric dipole moment variations. The calculated spectra were compared with the available experimental data. It was found that the torsional splittings and frequencies are relatively well reproduced, whereas the wagging and bending frequencies are slightly too high. Copyright 1998 Academic Press. PMID- 9753558 TI - Isotope Substitution in near Local Mode Molecules: Bending Overtones nnu2 (n = 2, 3) of the HDS Molecule. AB - There are two main goals in the present study: (a) to record and analyze the weak overtones nnu2 (n = 2, 3) of the bending fundamental nu2 of the HDS molecule and (b) to derive new isotopic relations for spectroscopic parameters applicable to predict, at least qualitatively, the structures of the excited bending states of HDX-type molecules. Copyright 1998 Academic Press. PMID- 9753559 TI - Analysis and Transition Probabilities for the Na2 B1Piu --> X1Sigma+g System Using as Excitation the 4727-A Ar+ Laser Line. AB - The fluorescence spectrum of Na2 induced by the 4726.87 A line of an Argon ion laser has been analyzed with special emphasis on determination of accurate relative intensities. This work extends previous observations on Na2 by using this excitation line. We have observed eight fluorescence series for the B1Piu - > X1Sigma+g band system corresponding to the excitation transitions v' = 9, J' = 38 <-- v" = 1, J" = 37; v' = 14, J' = 46 <-- v" = 4, J" = 47; v' = 14, J' = 81 <- v" = 3, J" = 80; v' = 15, J' = 65 <-- v" = 4, J" = 66; v' = 12, J' = 36 <-- v" = 3, J" = 36; v' = 12, J' = 40 <-- v" = 3, J" = 39 v' = 17, J' = 45 <-- v" = 6, J' = 45 and v' = 18, J' = 63 <-- v" = 6, J" = 63. The seven latter series are reported for the first time. Optically pumped laser transitions obtained in Na2 vapor by using this excitation wavelength have been assigned. The radiative transition probabilities for the observed fluorescence series were calculated using hybrid potential energy curves for the B1Piu and X1Sigma+g states constructed up to last vibrational levels and an ab initio transition dipole moment function. Radiative lifetimes for the rovibrational levels of the upper states pumped by the laser line have also been calculated. The transition probabilities and lifetimes agree with the corresponding observed measurements, usually within the experimental uncertainty. From the rotational satellite structure with DeltaJ' = +/-1 and +/-2 of some bands, for the most intense fluorescence series, collision-induced transition rates and average cross sections have been obtained. Copyright 1998 Academic Press. PMID- 9753560 TI - Line Broadening in the Fundamental Band of CO in CO-He and CO-Ar Mixtures. AB - We present accurate broadening coefficients for many lines of the fundamental band of CO highly diluted in He and Ar at 301.5 K. It is shown that a soft collision model gives a reasonable fit to the data at pressures up to 1 atm. A distinction is drawn between the accuracy with which fitting parameters may be determined and the accuracy with which a spectral profile fits experimental data. Differences between a model profile and the experimental profile are quantified, and the underlying physics omitted from the spectral model is briefly discussed. Copyright 1998 Academic Press. PMID- 9753561 TI - Ab Initio Determination of the Roto-Torsional Energy Levels of trans-1,3 Butadiene. AB - In this paper, the flexible model based on relaxed ab initio calculations, which has been several times employed for vibrational calculations, is extended to the analysis of the rotational structures starting by the roto-torsional bands of trans-1,3-butadiene. For this purpose, the potential energy surface and the kinetic energy parameters of the nu13 vibrational mode of butadiene are obtained with the Moller-Plesset perturbation theory up to the second order and the 6 31G(d, p), 6-31G(df, p), 6-311G(d, p), 6-311G(df, p), and 6-311G(df, pd) basis sets. The torsional levels of the -h6, -d4, and -d6 isotopic species are calculated variationally and are compared with experimental data. It may be concluded that the one-dimensional model appears sufficiently accurate for butadiene-h6 and -d4, whereas a large kinetic interaction with the lowest wagging mode is observed for butadiene-d6. The rotational levels corresponding to the first vibrational states of the -h6 and -d4 species are determined variationally up to J = 17 and J = 11 from the ab initio spectroscopic parameters which have been expanded as functions of the torsional coordinate using symmetry adapted series. The torsional wavefunction is contracted to reduce the size of the Hamiltonian matrix. A good agreement with the observed transitions is obtained for the first states v = 0 and v = 1. As is expected, the K doubling obtained is relatively small. For this reason, the quartic and sextic centrifugal distortion constants are obtained from the least-square fit of the variational levels to the perturbation theory equations for the symmetric top. Copyright 1998 Academic Press. PMID- 9753562 TI - Dispersed Fluorescence from the 460-nm Band System of NiCl2: Observation of a New, Low-Lying Electronic State. AB - The dispersed fluorescence spectrum of lines in the 460 nm band system of NiCl2 (nickel dichloride) is presented. The fluorescence was pumped on single rotational transitions of two spin components in a vibrational band observed previously in the laser excitation spectrum. Fluorescent transitions to levels of the &Xtilde;3Sigma-g (ground) state are assigned and discussed. The spectra also show clear evidence for a previously unidentified low-lying electronic state, assumed to be the A state, which lies about 2000 cm-1 above the ground state. This is likely to be of 3Pig character and is discussed on that basis. The wavenumber of the symmetric stretching vibration of NiCl2 in the A state is determined to be 367 cm-1, slightly larger than the &Xtilde; state value (360 cm 1). Copyright 1998 Academic Press. PMID- 9753563 TI - Laser Excitation Spectroscopy of the &Btilde;2A1-&Xtilde;2A1 Transition of the CaOCH3 Radical. AB - The (0-0) band of the &Btilde;2A1- &Xtilde;2A1 transition of CaOCH3 at 565 nm has been recorded and analyzed at Doppler limited resolution (FWHM = 0.005 cm-1). The molecule was generated using a laser ablation/molecular beam source in which a solid calcium rod was ablated and a mixture of argon seeded with a few percent of methanol was used as the carrier gas. The rotational analysis of the band is consistent with a linear Ca-O-C geometry in both electronic states. Values for the spin-rotation parameters are determined for the excited &Btilde;2A1 state. Copyright 1998 Academic Press. PMID- 9753564 TI - Precise Molecular Constants for the 6Li2 A1Sigma+u-X1Sigma+g System by Sub Doppler Polarization Spectroscopy. AB - We report here new and more accurate molecular constants from sub-Doppler polarization spectroscopy of the A1Sigma+u-X1Sigma+g system of 6Li2 using single mode cw dye lasers. These new constants cover the range of vibrational levels from v" = 0-8 in the ground state and v' = 0-24 in the excited state. New molecular constants and RKR potential energy curves for the A1Sigma+u and X1Sigma+g states are given. The Te value for the A1Sigma+u state is 14068.043(34) cm-1. The analysis indicates that there is a noticeable breakdown of the Born Oppenheimer approximation for the 6Li2 and 7Li2 isotopomers. Copyright 1998 Academic Press. PMID- 9753565 TI - High-Resolution Analysis of the nu6, nu7, nu8, and nu9 Bands of H15N16O3 Measured by Fourier Transform Spectroscopy. AB - The analysis of the nu6, nu7, nu8, and nu9 bands of H15N16O3 located at 646.9641, 578.4719, 743.6166, and 458.2917 cm-1, respectively, has been carried out in the 400-800 cm-1 region using high-resolution Fourier transform spectra recorded at Ottawa. Using the ground state energy levels calculated from the v = 0 rotational constants of H15N16O3 [A. P. Cox, M. C. Ellis, C. J. Attfield, and A. C. Ferris, J. Mol. Struct. 320, 91-106 (1994)], it was possible to assign the A-type nu6 and nu7 bands and the C-type nu8 and nu9 bands of H15N16O3 up to high J and Ka rotational quantum numbers. The v6 = 1, v7 = 1, v8 = 1, and v9 = 1 experimental energy levels were then introduced in a least-squares fit calculation and precise upper state Hamiltonian constants (band centers and rotational constants) were determined allowing one to reproduce the infrared data to within the experimental uncertainty. Copyright 1998 Academic Press. PMID- 9753566 TI - Fourier Transform Spectroscopy of the nu3 and nu6 Bands of D3Si35Cl: Rovibrational Constants of the v = 0, v3 = 1, and v6 = 1 States. AB - The nu3 and nu6 infrared bands near 500 cm-1 of monoisotopic D3Si35Cl have been studied with a resolution of 3.3 x 10(-3) cm-1. More than 800 transitions of nu3 and 2600 of nu6 have been assigned and fitted to ground and excited state parameters, final sigma(Fit) = 2.93 x 10(-4) cm-1. Ground state parameters up to quartic terms have been obtained. The excited state model included l(2, 2) and l(2, -1) interactions within nu6 and Coriolis x, y resonance between nu3 and nu6, interaction parameters being determined with high significance in all cases. The band centers nuo were determined, nu3 = 538.502 cm-1 and nu6 = 491.260 cm-1. Excellent agreement of experimental parameters with those from the ab initio harmonic and anharmonic force fields is noted. The transition moment ratio, ||u3:u6 || = 1:0.6(1), was determined by band contour simulation. Copyright 1998 Academic Press. PMID- 9753567 TI - The Coriolis Interaction between the v2 = 1 and v3 = 2 States of Nitrosyl Bromide: Anomalous Quadrupole Patterns and Interstate Transitions in the Millimeter-Wave Spectrum. AB - The millimeter-wave rotational spectra of 79BrNO and 81BrNO in the v2 = 1 and v3 = 2 vibrational states have been reinvestigated. Measurements of the rotational spectrum in the region of maximum c-type Coriolis interaction between the two states allowed the previous analysis to be extended to account for some uncommon effects. For the most perturbed transitions the nuclear quadrupole hyperfine structure arises from coupling of not only the bromine nucleus, but also the nitrogen nucleus with the rotational angular momentum. These effects were satisfactorily fitted with a Hamiltonian describing Coriolis coupling in a molecule with two quadrupolar nuclei. The successful analysis of pure rotational transitions then allowed accurate prediction of rovibrational transitions, six of which were measured for 79BrNO and four for 81BrNO. Copyright 1998 Academic Press. PMID- 9753568 TI - Infrared Diode Laser Spectroscopy of Fundamental and Hot Bands of BBr (X1Sigma+). AB - Infrared absorption spectra of the four common isotopic forms of the transient molecule BBr (11B79Br, 11B81Br, 10B79Br, and 10B81Br) have been observed in natural abundance. The spectra were detected in the region between 650 and 720 cm 1 using diode laser spectroscopy of a BBr3/He ac discharge. Over 150 lines consisting of both fundamental and hot bands up to v" = 5 have been fitted to Watson's isotopically invariant coefficients (Ukl). Spectroscopic parameters for each isotopomer could be derived from these coefficients; for 11B79Br, omegae = 685.1892(15) cm-1 and omegaexe = 3.73632(95) cm-1. The equilibrium dissociation energy obtained from Dunham's expanded Morse potential is 32 711(43) cm-1. Copyright 1998 Academic Press. PMID- 9753569 TI - High-Resolution Infrared Study of the nu14, nu17, and nu18 Bands of 11B2H6 and 10B11BH6. AB - Using high-resolution Fourier transform spectra, a thorough analysis of the nu14 c-type, nu17 a-type, and nu18 a-type bands of both 11B2H6 and 10B11BH6 has been carried in the 10.3-, 6.2-, and 8.5-um spectral regions, respectively. From this analysis a large set of precise ground state combination differences with J values up to 36 (31) and Ka values extending to 18 (18) was derived for 11B2H6(10B11BH6). These data were fitted using a Watson-type Hamiltonian leading to accurate ground state rotational constants. An rs value for the B-B distance has been determined to be 1.7645(10) A. The determination of upper state Hamiltonian constants proved to be much more difficult since the corresponding rotational levels of each of the bands are strongly perturbed by nearby dark states. To account for these strong localized resonances, it was necessary to introduce the relevant interacting terms in the Hamiltonian matrix. As a result it was possible to calculate the upper state energy levels quite satisfactorily. From these fits, estimates of the band centers and a few of the rotational constants of the resonating dark states were obtained. Copyright 1998 Academic Press. PMID- 9753570 TI - High-Resolution FTIR Spectrum of the nu5 Band of HCOOD. AB - The high-resolution Fourier transform infrared spectrum of HCOOD has been measured in the nu5 region between 1120 and 1220 cm-1 with a resolution of 0.004 cm-1. As expected for an in-plane vibrational fundamental mode, the nu5 band is a hybrid band consisting of both a-type and b-type transitions. Using the Watson's A-reduced Hamiltonian in the Ir representation, 1943 infrared transitions have been assigned and fitted to give 12 rovibrational constants for the v5 = 1 state. The nu5 band is primarily A type with a band center at 1177.09378 +/- 0.00002 cm 1. It is found that nu5 is slightly perturbed by the nearby 2nu7. About 90 perturbed transitions were identified. Copyright 1998 Academic Press. PMID- 9753571 TI - Fourier Transform Infrared Spectroscopy and Vibrational Coupling in the OH Bending Band of 13CH3OH. AB - We present in this work a high-resolution Fourier transform infrared study of the OH-bending vibrational band of 13CH3OH. We have investigated the 1070-1400 cm-1 spectral region at 0.002 cm-1 resolution using the modified Bomem DA3.002 Fourier transform spectrometer at the Steacie Institute for Molecular Sciences at the National Research Council of Canada in Ottawa. This study has led to (i) determination of excited-state J(J + 1) subband expansion coefficients and (ii) characterization of a variety of interactions coupling the different vibrational modes, notably a strong Fermi resonance between the OH bend and the torsionally excited CH3-rocking mode. The OH-bending band is widely spread with Q subbranches grouped in two peaks at about 1312 and 1338 cm-1. The lower levels for all assigned subbands were confirmed using closed loops of IR and FIR transitions. The subbands have been fitted to J(J + 1) power-series expansions in order to obtain the subband origins and the state-specific energy expansion coefficients for both the OH-bending and excited torsional CH3-rocking states. The strong interaction between the OH-bending state and the first excited torsional CH3 rocking state gives rise to several "extra" forbidden subbands due to intensity borrowing. The asymmetry splitting of the (ntauK) v = (122)OH A OH-bending doublet was found to be anomalously small, and the splitting of the (122)rA CH3 rocking doublet is observed to be enhanced. We have identified a network of intermode interactions causing this unusual behavior, but a quantitative analysis of the vibrational coupling is restricted by limited knowledge of the unperturbed positions of the interacting levels. All these interactions provide relaxation channels for intramolecular vibrational redistribution among the lower vibrational modes in 13CH3OH. Another important finding is that the torsion-K rotation energy curves in the OH-bending state display an inverted pattern compared to the ground state. Copyright 1998 Academic Press. PMID- 9753572 TI - The Far Infrared Spectrum of HOCl: Line Positions and Intensities. AB - The far infrared spectrum of HOCl has been recorded at high resolution between 20 and 360 cm-1 by means of Fourier transform spectroscopy, and it was possible to observe pure rotation lines involving rotational levels with high Ka quantum numbers (up to Ka = 9). These lines combined with microwave and tunable far infrared data available in the literature were least squares fitted using a Watson-type Hamiltonian. The fitting leads to precise sets of rotational and centrifugal distortion constants for the ground states of both isotopomers HO35Cl and HO37Cl. Also relative line intensities were measured and their fitting allowed the determination of rotational corrections to the b-component of the permanent transition moment. Finally, to get Hamiltonian constants consistent with the newly determined ground state constants for the (100), (010), (001) vibrational states, available data concerning the nu1, nu2, and nu3 bands were refitted. Three interesting points are to be stressed. For the (001) state, we were able to complete the existing data with rotation lines observed in our spectra up to rather high Ka values (Ka = 7). For HO35Cl, we were able to show that some (010) and (100) levels are perturbed by levels of the (002) and (030) states, respectively, through Coriolis-type interactions. This allows the determination of the band centers of these two dark states. Copyright 1998 Academic Press. PMID- 9753573 TI - The Molecular Constants of 12CH3I in the Ground and v6 = 1 Excited Vibrational State. AB - A set of 26 new measurements have been recorded by a Doppler-free double resonance technique, with a relative accuracy of about 10(-8). These data are combined to previous FTIR measurements to refine the molecular constants of 12CH3I. All the available literature data have been revisited, a few assignments updated, and some frequencies changed according to new laser standards. A simultaneous fit to all the measurements of infrared, microwave, and radiofrequency transitions produced an improved set of parameters, independent of previously determined constants. Copyright 1998 Academic Press. PMID- 9753575 TI - Line Intensities and Self-Broadening Coefficients for Methane Lines between 5500 and 6180 cm-1 Retrieved with a Multispectrum Fitting Technique. PMID- 9753574 TI - Microwave Spectroscopy of BrBO. AB - The microwave spectra of the four isotopomers of the BrBO molecule (79Br10BO, 81Br10BO, 79Br11BO, and 81Br11BO) were observed in a dc glow discharge plasma of a mixture of boron tribromide vapor and oxygen gas. Rotational transitions of BrBO were measured for the ground state as well as for the vibrationally excited states, nu2 (bend) and nu3 (Br-B str.), for all of four isotopomers, and associated 2nu2, nu2 + nu3, and 2nu3 for the 11B species. The l = 0 substate of the 2nu2 state interacts with the nu3 state through the Fermi resonance. The rotational constants determined for the ground states of the four isotopomers yield the substitution structure, rs(Br&sbond;B) = 1.835791(70) A and rs(B&dbond;O) = 1.20472(25) A. The pi character and ionic character of the Br-B bond, which are estimated from the bromine quadrupole coupling constant eQq, are discussed through the comparison with those of related molecules such as BrCN and BrBS. Copyright 1998 Academic Press. PMID- 9753576 TI - The Far-Infrared Spectrum of Deuterium Iodide. PMID- 9753577 TI - Predissociation of the 7Li2 2(3)Sigma+g State. PMID- 9753578 TI - Quadrupole Coupling Constants for 33SO3: Microwave Measurements for Ar-33SO3 and ab Initio Results for the 33SO3 Monomer. PMID- 9753579 TI - The Number of Harmonic Frequencies of a Symmetrical Molecule: A Correction. PMID- 9753582 TI - Introduction PMID- 9753583 TI - Proceedings of the 6th conference on the neurobiology of learning and memory. Brain and memory: from genes to behavior. Some historical background of topics in this conference. PMID- 9753584 TI - Mapping brain networks engaged by, and changed by, learning. AB - Major goals of research into the neurobiology of learning and memory are to identify (1) brain areas/circuitries that subserve different mnemonic functions and (2) chemistries that encode the memory trace. The discovery that activity modulates neuronal gene expression provided techniques attendant to the first goal and candidates for cellular changes pertinent to the second. Studies in our laboratories have exploited activity-regulated changes in c-fos gene expression to map regions engaged in two-odor discrimination learning, with particular interest in neuronal groups in hippocampus and amygdala. The results of these studies demonstrate that the subdivisions of hippocampus and amygdala do not act in concert across behaviors but are differentially activated depending on task demands. In hippocampus, preferential activation of field CA3 was uniquely associated with initial learning of an odor pair, whereas predominant activation of CA1 occurred with exploration of a novel field and with overtrained responding to odors. The reappearance of precisely the same balance of subfield activation within disparate behavioral contexts was taken to suggest that the hippocampus has basic modes of function that recur in different circumstances and make rather generalized contributions to behavior. Within the amygdala, the basolateral division was most prominently active during task acquisition but not during performance of the well-learned discrimination. Indeed, the amygdala appeared to play the dominant role relative to hippocampus in the early stages of associating positive and negative valences with discriminative cues. These results demonstrate that the balance of neuronal activity both within and between limbic structures changes across sequential stages of odor learning in a fashion that is likely to define behavioral output. PMID- 9753585 TI - Immediate-early genes and synaptic function. AB - Classical studies have demonstrated a role for protein synthesis in long-term memory. The focus of our research is to identify the proteins that are essential for memory and to discover how they contribute to activity-dependent neuronal plasticity. We have developed whole-animal models that maximize the induction of activity-dependent genes and have used differential cloning techniques to identify a set of novel, neuronal immediate-early genes (IEGs). Neuronal IEGs encode transcription factors, cytoskeletal proteins, growth factors, metabolic enzymes, and proteins involved in signal transduction. The biochemical and cell biological properties of these molecules provide important insights into mechanisms that contribute to neuronal plasticity. Recently, we identified a subset of IEGs that appear to function at the synapse. These molecules extend the functional repertoire of IEGs and may provide insight into how IEGs can contribute to synapse-specific plasticity. PMID- 9753586 TI - Molecular, cellular, and neuroanatomical substrates of place learning. AB - Learning and remembering the location of food resources, predators, escape routes, and immediate kin is perhaps the most essential form of higher cognitive processing in mammals. Two of the most frequently studied forms of place learning are spatial learning and contextual conditioning. Spatial learning refers to an animal's capacity to learn the location of a reward, such as the escape platform in a water maze, while contextual conditioning taps into an animal's ability to associate specific places with aversive stimuli, such as an electric shock. Recently, transgenic and gene targeting techniques have been introduced to the study of place learning. In contrast with the abundant literature on the neuroanatomical substrates of place learning in rats, very little has been done in mice. Thus, in the first part of this article, we will review our studies on the involvement of the hippocampus in both spatial learning and contextual conditioning. Having demonstrated the importance of the hippocampus to place learning, we will then focus attention on the molecular and cellular substrates of place learning. We will show that just as in rats, mouse hippocampal pyramidal cells can show place specific firing. Then, we will review our evidence that hippocampal-dependent place learning involves a number of interacting physiological mechanisms with distinct functions. We will show that in addition to long-term potentiation, the hippocampus uses a number of other mechanisms, such as short-term-plasticity and changes in spiking, to process, store, and recall information. Much of the focus of this article is on genetic studies of learning and memory (L&M). However, there is no single experiment that can unambiguously connect any cellular or molecular mechanism with L&M. Instead, several different types of studies are required to determine whether any one mechanism is involved in L&M, including (i) the development of biologically based learning models that explain the involvement of a given mechanism in L&M, (ii) lesion experiments (genetics and pharmacology), (iii) direct observations during learning, and (iv) experiments where learning is triggered by turning on the candidate mechanism. We will show how genetic techniques will be key to unraveling the molecular and cellular basis of place learning. PMID- 9753587 TI - NMDA receptor-dependent and metabotropic glutamate receptor-dependent forms of long-term depression coexist in CA1 hippocampal pyramidal cells. AB - We have found that two distinct forms of long-term depression (LTD), one dependent on the activation of NMDA receptors (NMDARs) and the other dependent on the activation of metabotropic glutamate receptors (mGluRs), coexist in pyramidal cells of the CA1 region of the hippocampus of juvenile rats (11-35 days). Both forms were pathway specific, required membrane depolarization, and were blocked by chelating postsynaptic Ca2+ with BAPTA. The mGluR-LTD, but not the NMDAR-LTD, was blocked by the T-type Ca2+ channel blocker Ni2+ and intracellular administration of a protein kinase C inhibitory peptide. In contrast, the protein phosphatase inhibitor Microcystin LR blocked NMDAR-LTD, but not mGluR-LTD. NMDAR LTD is associated with a decrease in the size of quantal excitatory postsynaptic currents, whereas for mGluR-LTD there was no change in quantal size, but a large decrease in the frequency of events. While mGluR-LTD did not interact with NMDAR dependent long term potentiation (LTP), NMDAR-LTD was capable of reversing LTP. Prior saturation of mGluR-LTD had no effect on NMDAR-LTD. NMDAR-LTD and mGluR-LTD thus appear to be mechanistically distinct forms of synaptic plasticity in that they share neither induction nor expression mechanisms. PMID- 9753588 TI - Contributions of cell adhesion molecules to altered synaptic weightings during memory consolidation. AB - The fundamental concept that synapse growth and change are associated with learning is considered a "replay" of early neurodevelopmental principles that instruct neural connectivity pattern. Common mechanisms suggested to link the process of memory formation through synaptic elaboration are exemplified by the activity of cell adhesion molecules following learning and that center on waves of glycoprotein synthesis occurring in the 6- to 8-h and 10- to 12-h posttraining periods of consolidation. These are associated with spatially clustered granule cells in the adult rat hippocampus that show a transient time-dependent increase in ribosome production and greater microtubular complexity and dendritic spine number 6 to 8 h following training. The elimination and/or selection of the synapses to be retained in the memory trace is proposed to be dependent on cell adhesion molecule glycosylation events in the 10- to 12-h posttraining period. The existence of similar cell adhesion molecule glycosylation mechanisms within a corticohippocampal pathway is used to contribute to a model of memory in which sensory representations are eventually consolidated through relative change in synaptic weightings. PMID- 9753589 TI - Memory and the brain: unexpected chemistries and a new pharmacology. AB - Efforts to characterize long-term potentiation (LTP) and to identify its substrates have led to the discovery of novel synaptic chemistries, computational algorithms, and, most recently, pharmacologies. Progress has also been made in using LTP to develop a "standard model" of how unusual, but physiologically plausible, levels of afferent activity create lasting changes in the operating characteristics of synapses in the cortical telencephalon. Hypotheses of this type typically distinguish induction, expression, and consolidation stages in the formation of LTP. Induction involves a sequence consisting of theta-type rhythmic activity, suppression of inhibitory currents, intense synaptic depolarization, NMDA receptor activation, and calcium influx into dendritic spines. Calcium dependent lipases, kinases, and proteases have been implicated in LTP induction. Regarding the last group, it has been recently reported that theta pattern stimulation activates calpain and that translational suppression of the protease blocks potentiation. It is thus likely that proteolysis is readily driven by synaptic activity and contributes to structural reorganization. LTP does not interact with treatments that affect transmitter release, has a markedly differential effect on the currents mediated by colocalized AMPA vs NMDA synaptic receptors, changes the waveform of the synaptic current, modifies the effects of drugs that modulate AMPA receptors, and is sensitive to the subunit composition of those receptors. These results indicate that LTP is expressed by changes in AMPA receptor operations. LTP is accompanied by modifications in the anatomy of synapses and spines, something which accounts for its extreme duration (weeks). As with various types of memory, LTP requires about 30 min to consolidate (become resistant to disruption). Consolidation involves adhesion chemistries and, in particular, activation of integrins, a class of transmembrane receptors that control morphology in numerous cell types. Platelet activating factor and adenosine may contribute to consolidation by regulating the engagement of latent integrins. How consolidation stabilizes LTP expression is a topic of intense investigation but probably involves modifications to one or more of the following: membrane environment of AMPA receptors; access of regulatory proteins (e.g., kinases, proteases) to the receptors; receptor clustering; and space available for receptor insertion. Attempts to enhance LTP have focused on the induction phase and resulted in a class of centrally active drugs ("ampakines") that positively modulate AMPA receptors. These compounds promote LTP in vivo and improve the encoding of variety of memory types in animals. Positive results have also been obtained in preliminary studies with humans. PMID- 9753590 TI - Information processing with frequency-dependent synaptic connections. AB - The efficacy of synaptic transmission between two neurons changes as a function of the history of previous activations of the synaptic connection. This history dependence can be characterized by examining the dependence of transmission on the frequency of stimulation. In this framework synaptic plasticity can also be examined in terms of changes in the frequency dependence of transmission and not merely in terms of synaptic strength which constitutes only a linear scaling mechanism. Recent work shows that the frequency dependence of transmission determines the content of information transmitted between neurons and that synaptic modifications can change the content of information transmitted. Multipatch-clamp recordings revealed that the frequency dependence of transmission is potentially unique for each synaptic connection made by a single axon and that the class of pre-postsynaptic neuron determines the class of frequency dependence (activity independent), while the unique activity relationship between any two neurons could determine the precise values of the parameters within a specific class (activity dependent). The content of information transmitted between neurons is also formalized to provide synaptic transfer functions which can be used to determine the role of the synaptic connection within a network of neurons. It is proposed that deriving synaptic transfer functions is crucial in order to understand the link between synaptic transmission and information processing within networks of neurons and to understand the link between synaptic plasticity and learning and memory. PMID- 9753591 TI - Synaptic plasticity and learning and memory: 15 years of progress. AB - Much has been learned over the past 15 years about the mechanisms of synaptic plasticity and their relationships to learning and memory processes. Some of the questions raised 15 years ago have been answered while others still remain elusive. This brief review attempts to evaluate the progress accomplished in this field and discusses four specific issues: (i) the relationships between mechanisms of synaptic plasticity and memory types, (ii) the relationships between stabilization of synaptic modifications and memory consolidation, (iii) the links between gene regulation and regulation of synaptic efficacy, and (iv) the relationships between synaptic dynamics and synaptic plasticity. Although it is relatively easy to identify areas in which progress has been made, it is also clear that many areas remain highly controversial and will keep neuroscientists busy for years to come. PMID- 9753593 TI - Differential roles of the frontal cortex, basal ganglia, and cerebellum in visuomotor sequence learning. PMID- 9753592 TI - The basal ganglia and chunking of action repertoires. AB - The basal ganglia have been shown to contribute to habit and stimulus-response (S R) learning. These forms of learning have the property of slow acquisition and, in humans, can occur without conscious awareness. This paper proposes that one aspect of basal ganglia-based learning is the recoding of cortically derived information within the striatum. Modular corticostriatal projection patterns, demonstrated experimentally, are viewed as producing recoded templates suitable for the gradual selection of new input-output relations in cortico-basal ganglia loops. Recordings from striatal projection neurons and interneurons show that activity patterns in the striatum are modified gradually during the course of S-R learning. It is proposed that this recoding within the striatum can chunk the representations of motor and cognitive action sequences so that they can be implemented as performance units. This scheme generalizes Miller's notion of information chunking to action control. The formation and the efficient implementation of action chunks are viewed as being based on predictive signals. It is suggested that information chunking provides a mechanism for the acquisition and the expression of action repertoires that, without such information compression would be biologically unwieldy or difficult to implement. The learning and memory functions of the basal ganglia are thus seen as core features of the basal ganglia's influence on motor and cognitive pattern generators. PMID- 9753594 TI - The nature of reinforcement in cerebellar learning. AB - In a now classic study, W. J. Brogden and W. H. Gantt (1942, American Journal of Physiology, 119, 277-278) demonstrated that movements (limbs, head, eyelid) elicited by direct electrical stimulation of certain regions of the cerebellum (particularly the ansiform lobe) could be trained to respond to neutral auditory or visual conditioned stimuli with appropriate pairing. In recent work we have replicated these results in detail and presented considerable evidence that the reinforcing or "teaching" pathway so activated for the learning of discrete movements is the inferior olive-climbing fiber projection system to the cerebellum. Very strong evidence now indicates that the memory traces for this "skilled response" learning are formed and stored in the cerebellum. The climbing fiber system and inhibitory pathway from the interpositus nucleus to the inferior olive appear to form a neural instantiation of the Resorla-Wagner formulation of classical conditioning and accounts for the "cognitive" phenomenon of blocking. It is concluded that reinforcement in this form of learning is not due simply to contiguity/contingency or to unconditioned stimulus aversiveness, per se, but rather to activation of a particular brain circuit, here the climbing fiber system, a circumstance that may apply to other forms of learning, with other reinforcement circuits, as well. PMID- 9753595 TI - A role for the cerebellum in learning movement coordination. AB - We have examined several different paradigms of adaptation and of "acquisition of skill"-skill defined as a movement specialized to meet a certain goal and gained through practice. In each paradigm, change occurs through trial-and-error performance. In some of the tasks, damage of cerebellar cortex impairs adaptation and not performance. The deficits in performance cannot explain the deficits in adaptation. In some of the tasks, the discharge of Purkinje cells and, by inference, the discharge of inferior olive cells and mossy fibers have occurred in a manner consistent with the Marr-Albus theory of motor learning. We extend the theory to show how parallel fibers could implement both the coordination of complex movements and the learning of new movements. The size of the response combinations would be proportionate to the length of parallel fibers. The mechanism proposed here would permit optimized complex movement behaviors to respond to specific behavioral contexts rapidly, stereotypically, and automatically. The mechanism would permit storage of many context-response couplings and many complex responses. The mechanism would permit privacy, individuality, and a large number of behavioral responses. PMID- 9753596 TI - A comparative perspective on motor learning. PMID- 9753597 TI - Consolidation of visual associative long-term memory in the temporal cortex of primates. AB - Neuropsychological theories have proposed a critical role for the interaction between the medial temporal lobe and the neocortex in the formation of long-term memory for facts and events, which has often been tested by learning of a series of paired words or figures in humans. We have examined neural mechanisms underlying the memory "consolidation" process by single-unit recording and molecular biological methods in an animal model of a visual pair-association task in monkeys. In our previous studies, we found that long-term associative representations of visual objects are acquired through learning in the neural network of the anterior inferior temporal (IT) cortex. In this article, we propose the hypothesis that limbic neurons undergo rapid modification of synaptic connectivity and provide backward signals that guide the reorganization of neocortical neural circuits. Two experiments tested this hypothesis: (1) we examined the role of the backward connections from the medial temporal lobe to the IT cortex by injecting ibotenic acid into the entorhinal and perirhinal cortices, which provided massive backward projections ipsilaterally to the IT cortex. We found that the limbic lesion disrupted the associative code of the IT neurons between the paired associates, without impairing the visual response to each stimulus. (2) We then tested the first half of this hypothesis by detecting the expression of immediate-early genes in the monkey temporal cortex. We found specific expression of zif268 during the learning of a new set of paired associates in the pair-association task, most intensively in area 36 of the perirhinal cortex. All these results with the visual pair-association task support our hypothesis and demonstrate that the consolidation process, which was first proposed on the basis of clinico-psychological evidence, can now be examined in primates using neurophysiolocical and molecular biological approaches. PMID- 9753598 TI - Cell-assembly coding in several memory processes. AB - The present paper discusses why the cell assembly, i.e., an ensemble population of neurons with flexible functional connections, is a tenable view of the basic code for information processes in the brain. The main properties indicating the reality of cell-assembly coding are neurons overlaps among different assemblies and connection dynamics within and among the assemblies. The former can be detected as multiple functions of individual neurons in processing different kinds of information. Individual neurons appear to be involved in multiple information processes. The latter can be detected as changes of functional synaptic connections in processing different kinds of information. Correlations of activity among some of the recorded neurons appear to change in multiple information processes. Recent experiments have compared several different memory processes (tasks) and detected these two main properties, indicating cell assembly coding of memory in the working brain. The first experiment compared different types of processing of identical stimuli, i.e., working memory and reference memory of auditory stimuli. The second experiment compared identical processes of different types of stimuli, i.e., discriminations of simple auditory, simple visual, and configural auditory-visual stimuli. The third experiment compared identical processes of different types of stimuli with or without temporal processing of stimuli, i.e., discriminations of elemental auditory, configural auditory-visual, and sequential auditory-visual stimuli. Some possible features of the cell-assembly coding, especially "dual coding" by individual neurons and cell assemblies, are discussed for future experimental approaches. PMID- 9753600 TI - The neurophysiology of reminiscence. PMID- 9753599 TI - Physiological memory in primary auditory cortex: characteristics and mechanisms. AB - "Physiological memory" is enduring neuronal change sufficiently specific to represent learned information. It transcends both sensory traces that are detailed but transient and long-term physiological plasticities that are insufficiently specific to actually represent cardinal details of an experience. The specificity of most physiological plasticities has not been comprehensively studied. We adopted receptive field analysis from sensory physiology to seek physiological memory in the primary auditory cortex of adult guinea pigs. Receptive fields for acoustic frequency were determined before and at various retention intervals after a learning experience, typified by single-tone delay classical conditioning, e.g., 30 trials of tone-shock pairing. Subjects rapidly (5-10 trials) acquire behavioral fear conditioned responses, indexing acquisition of an association between the conditioned and the unconditioned stimuli. Such stimulus-stimulus association produces receptive field plasticity in which responses to the conditioned stimulus frequency are increased in contrast to responses to other frequencies which are decreased, resulting in a shift of tuning toward or to the frequency of the conditioned stimulus. This receptive field plasticity is associative, highly specific, acquired within a few trials, and retained indefinitely (tested to 8 weeks). It thus meets criteria for "physiological memory." The acquired importance of the conditioned stimulus is thought to be represented by the increase in tuning to this stimulus during learning, both within cells and across the primary auditory cortex. Further, receptive field plasticity develops in several tasks, one-tone and two-tone discriminative classical and instrumental conditioning (habituation produces a frequency-specific decrease in the receptive field), suggesting it as a general process for representing the acquired meaning of a signal stimulus. We have proposed a two-stage model involving convergence of the conditioned and unconditioned stimuli in the magnocellular medial geniculate of the thalamus followed by activation of the nucleus basalis, which in turn releases acetylcholine that engages muscarinic receptors in the auditory cortex. This model is supported by several recent findings. For example, tone paired with NB stimulation induces associative, specific receptive field plasticity of at least a 24-h duration. We propose that physiological memory in auditory cortex is not "procedural" memory, i.e., is not tied to any behavioral conditioned response, but can be used flexibly. PMID- 9753601 TI - Distributed memory for both short and long term. AB - Neuropsychology points to the wide distribution of cortical memory networks. Electrophysiology and neuroimaging indicate that working memory, like long-term memory, is a widely distributed function, largely neocortical. Most of the evidence available from those three methodologies suggests that both working memory and long-term memory share the same substrate: a system of broad, partly overlapping and interconnected neocortical networks. Working memory appears mostly, if not completely, characterized by the sustained activation of one widely distributed network of long-term memory. That activation is at least in part sustained by reentrant excitatory loops through the different neuronal assemblies that constitute the network and that represent the associated features of the memorandum. PMID- 9753602 TI - Images of medial temporal lobe functions in human learning and memory. AB - Investigations of the neural basis of mammalian memory have focused more often on the medial temporal lobe (MTL) than on any other brain region. In humans, the amnesic syndrome revealed the essential importance of the multiple structures located in the MTL system for declarative memory (the remembrance of events and facts). Other neural systems mediate procedural forms of memory, including delay eyeblink conditioning, which depends on the cerebellum, and cognitive skill learning, which depends on the striatum. We review three functional imaging studies that reveal different patterns of MTL activation associated with declarative and procedural memory tasks. One study shows separate MTL activations during the encoding or retrieval of declarative memories. A second study shows MTL activation that occurs in parallel with cerebellum-dependent delay eyeblink conditioning, but does not appear to influence that form of procedural memory. A third study reveals suppression of the MTL during striatum-dependent cognitive skill learning. These studies provide images of MTL activations that are correlated with, independent from, or antagonistic to memory performance. PMID- 9753604 TI - A festschrift in honor of dr. James L. McGaugh PMID- 9753603 TI - On the relations among priming, conscious recollection, and intentional retrieval: evidence from neuroimaging research. AB - Neurobiological distinctions among forms of memory have been investigated mainly from the perspective of lesion studies in nonhuman animals and experiments with human neurological patients. We consider recent neuroimaging studies of healthy human volunteers using positron emission tomography (PET) and functional magnetic resonance imaging (fMRI) that provide new information concerning the neural correlates of particular forms of memory retrieval. More specifically, we consider evidence indicating that priming, a form of implicit retrieval, is associated with decreased activity in various cortical regions. We also consider evidence suggesting that two components of explicit retrieval-intentional or effortful search and successful conscious recollection-are preferentially associated with increased activity in prefrontal and medial temporal regions, respectively. PMID- 9753605 TI - Transmissible spongiform encephalopathies. AB - Scrapie, bovine spongiform encephalopathy (BSE), and the Creutzfeldt-Jakob disease (CJD) belong to a group of lethal neurodegenerative disorders in mammals. Prion diseases or transmissible spongiform encephalopathies (TSEs) are characterized by the accumulation of an abnormal isoform (PrPSc) of the host encoded cellular prion protein (PrPC) in the brain. The infectious agent, the 'prion,' is believed to be devoid of informational nucleic acid and to consist largely, if not entirely, of PrPSc. The PrP isoforms contain identical amino acid sequences yet differ in their overall secondary structure with the PrPSc isoform possessing a higher beta-sheet and lower alpha-helix content than PrPC. Elucidation of the three-dimensional structure of PrPC has provided important clues on the molecular basis of inherited human TSEs and on the species barrier phenomenon of TSEs. Nevertheless, the molecular mechanism of the conformational rearrangement of PrPC into PrPSc is still unknown, mainly due to the lack of detailed structural information on PrPSc. Within the framework of the 'protein only' hypothesis, two plausible models for the self-replication of prions have been suggested, the conformational model and the nucleation-dependent polymerization model. PMID- 9753606 TI - Cell growth activity of growth-blocking peptide. AB - Growth-blocking peptide (GBP) is an insect biogenic peptide that retards the development of lepidopteran larvae. cDNAs encoding GBP were isolated from three lepidopteran species: Pseudaletia separata, Mamestra brassicae and Spodoptera litura. Comparison of these molecules revealed that the GBP coding regions were 70% homologous. In contrast, the upstream regions of the deduced propeptides were only 33% identical. Sequence analysis further suggested that GBP shares some structural similarities with human epidermal growth factors. Bioassay data revealed that several pmol/ml of GBP stimulated DNA synthesis of a human keratinocyte cell line and of SF-9 insect cells. However, several nmol/ml of GBP did not stimulate cell proliferation at all. In vivo studies similarly indicated that low concentrations of GBP stimulated larval growth while high concentrations of GBP retarded growth. These data suggest that GBP acts as a growth factor that regulates insect larval development. PMID- 9753607 TI - MAP kinase pathways as a route for regulatory mechanisms of IL-10 and IL-4 which inhibit COX-2 expression in human monocytes. AB - Mitogen-activated protein kinases (MAPKs) are activated by various extracellular stimuli and play an important role in regulating the expression of proinflammatory molecules in monocytes/macrophages. We first questioned whether MAPK activation in involved in cyclooxygenase (COX)-2 expression in lipopolysaccharide (LPS)-stimulated human monocytes. LPS induced the expression of COX-2 protein and COX-2 mRNA as well as the phosphorylation and activation of extracellular signal-regulated protein kinase (ERK)2 and p38 MAPK in monocytes. The induction of COX-2 mRNA, COX-2 protein, and prostaglandin (PG)E2 by LPS was inhibited by the specific inhibitors of ERK and p38 MAPK, suggesting that the activation of ERK2 and p38 MAPK is involved in COX-2 expression in LPS-stimulated monocytes. Since we previously showed that interleukin (IL)-10 and IL-4 similarly inhibited COX-2 expression in LPS-stimulated monocytes, we next questioned whether these cytokines regulate the phosphorylation and activation of ERK2 and p38 MAPK in LPS-stimulated monocytes. Interestingly, LPS-induced phosphorylation and activation of ERK2 was significantly inhibited by IL-4 and IL-10, while that of p38 MAPK was inhibited by IL-10, but not IL-4. These results suggest that the mechanisms of inhibition by IL-10 and IL-4 of the LPS-induced expression of proinflammatory molecules could be ascribed to the regulatory effects of both cytokines on MAPK activation. PMID- 9753608 TI - Uridine diphosphoglucose dehydrogenase regulates proteoglycan expression: cDNA cloning and antisense study. AB - Using a reverse-transcriptase-polymerase chain reaction approach human and murine UDPG-dehydrogenase (GDH) was cloned from fibroblast mRNAs. Human enzyme is 97% and 27% identical with its murine and E. coli orthologs. Murine mRNA of 3.1 kb size is expressed in all the tissue studied at a level independent of glyceraldehyde-3-phosphate dehydrogenase (GADPH) mRNA. In human fibroblast in vitro, 2 GDH transcripts were observed. They were expressed proportionally to GAPDH. The simple pattern of human GDH Southern blotting suggests a single copy gene. An antisense oligonucleotide directed to the ATG region of the human enzyme inhibited 35S-sulphate incorporation into extracellular macromolecules, especially proteoglycans. These data indicate that GDH expression may regulate proteoglycan synthesis in the cells. PMID- 9753609 TI - Protein-protein interactions of the yeast Golgi t-SNARE Sed5 protein distinct from its neural plasma membrane cognate syntaxin 1. AB - Targeting of vesicles to the acceptor membrane in protein transport depends on membrane proteins called SNAREs. Saccharomyces cerevisiae Golgi t-SNARE Sed5 protein and its neural cognate syntaxin 1 have similar three alpha-helices which are predicted to form coiled coils. We dissected the helices of Sed5 and found several characteristics unexpectedly distinct from those of syntaxin 1. Most importantly, only the N-terminal helix is responsible for the binding of Sly1 protein while almost the entire molecule of syntaxin is necessary for the binding of the cognate, Munc-18. The N-terminal region of Sed5 protein also binds to the C-terminal helix and Sly1 protein interfered this binding. PMID- 9753610 TI - Attracted or repelled?--a matter of two neurons, one pheromone binding protein, and a chiral center. AB - Two species of scarab beetles, the Osaka beetle (Anomala osakana) and the Japanese beetle (Popillia japonica), utilize the opposite enantiomers of japonilure, (Z)-5-(1-decenyl)oxacyclopentan-2-one, as their sex pheromones. Each species produces only one of the enantiomers that functions as its own sex pheromone and as a very strong behavioral antagonist for the other species. Using an integrated approach we tested whether the discrimination of these two opposite signals is due to selective filtering by pheromone binding proteins or whether it originates in the specificity of ligand-receptor interactions. We found that the antennae of each of these two scarab species contain only a single pheromone binding protein, which associates with both enantiomers to a similar extent. The single neuron recording technique, on the other hand, showed that both species possess olfactory receptor neurons, colocalized in one sensillum, extremely specific to either (R)- or (S)-japonilure. Therefore, pheromone binding proteins (PBPs) alone cannot perform the task of chiral discrimination; enantiomeric specificity must be achieved by the interaction of the pheromone or the appropriate ligand-PBP complex with membrane receptors. PMID- 9753611 TI - c-Src and phosphatidylinositol 3-kinase are involved in NGF-dependent tyrosine phosphorylation of Shc in PC12 cells. AB - The adaptor protein Shc exists in three isoforms; p46, p52, and p66, and is a key regulator of a variety of biological processes. Our previous studies have shown that the tyrosine kinase c-Src phosphorylates Shc in a phosphatidylinositol (PtdIns) 4,5-bisphosphate-dependent manner. Here we demonstrate that PtdIns 3,4,5 trisphosphate stimulates phosphorylation of Shc by c-Src. The phosphorylation is blocked by a glutathione S-transferase fusion protein containing Shc phosphotyrosine binding (PTB) domain or a phosphotyrosine-containing Shc PTB domain-binding peptide. In rat pheochromocytoma cell line PC12, nerve growth factor (NGF) stimulates tyrosine phosphorylation of both Triton-soluble and insoluble Shc which was maximal at 2-5 min after NGF treatment. We find that pretreatment of PC12 cells with the PtdIns 3-kinase inhibitor wortmannin or LY294002 results in almost half inhibition of the NGF-dependent tyrosine phosphorylation of only Triton-insoluble Shc. Similar inhibitory effect is observed with tyrosine kinase inhibitors genistein and PP1. Upon NGF stimulation, c-Src also becomes tyrosine-phosphorylated and accumulates in the Triton insoluble fraction. The c-Src events are insensitive to wortmannin but sensitive to genistein. These results suggest that coordinate action of PtdIns 3-kinase and/or PtdIns 3,4,5-trisphosphate and c-Src can function as positive regulator in tyrosine phosphorylation of Shc in vitro and in vivo. PMID- 9753612 TI - Osteoclastogenesis inhibitory factor suppresses osteoclast survival by interfering in the interaction of stromal cells with osteoclast. AB - Osteoclastogenesis inhibitory factor (OCIF) was originally identified as a factor inhibiting osteoclast (OC) formation. The number of OC in rats treated with OCIF decreased, suggesting that OCIF inhibits OC formation in vivo; however, it is also possible that OCIF affects the number of OC by promoting apoptosis of OC. To address this issue, the effects of OCIF on the survival of OC were examined using well established mouse culture systems. OCIF dose-dependently inhibited OC formation in mouse marrow cultures. Addition of OCIF during day 0-3 did not alter the peak levels of OC formation on day 7 and 8. However, the addition of OCIF during day 5 and thereafter resulted in the rapid decrease of the number of OC. OCIF inhibited the survival of OC formed in mouse marrow cultures in dose- and time-dependent manners. The involvement of stromal cells in OC survival was examined using crude and stromal cell-depleted OC populations. OCIF dramatically inhibited the survival of crude OC populations rich with stromal cells. However, in stromal cell-depleted OC populations, OC spontaneously decreased in the absence of OCIF and OCIF did not enhance the decrease further at least up to 48 h. Apoptotic OC were detected in detached cell populations treated with OCIF in mouse marrow cultures and a specific inhibitor for caspase-3 rescued the death of OC. OCIF mutant lacking the death domain homologous regions inhibited OC survival, though the potency was much less than native OCIF. Taken together, OCIF inhibited not only OC recruitment but also OC survival. OCIF inhibited OC survival by interfering the interaction of stromal cells with OC. PMID- 9753613 TI - Anti-autolysis of trypsin by modification of autolytic site Arg117. AB - In order to improve the stability of trypsin, an approach to knock out the autolytic site has been carried out in this investigation. Compared with trypsins from other species, the autolytic site Arg117-Val118 of rat trypsin is the most interesting candidate to work on. The Arg117 residue was designed to be deleted or replaced by other amino acid residues to destroy the autolytic site. With DNA site-directed mutagenesis method, one deletion mutant and several replacement mutants were selected. After expression and purification, the kinetic and anti autolytic properties of mutant trypsins were studied. No net charge difference of the trypsin molecules was observed by native PAGE analysis. Kinetic studies show that the activities of mutants vary from one another. R117L gives 32 times the activity of wild type trypsin while R117C has no detective activity. Among 8 selected mutants with characteristic properties, 7 of them give prolonged half life during anti-autolytic assay with the exception of R117M which is more sensitive to autolysis. PMID- 9753614 TI - GABA (gamma-amino-butyric acid) neurotransmission: identification and fine mapping of the human GABAB receptor gene. AB - GABA (gamma-amino-butyric acid) receptors are a family of proteins involved in the GABAergic neurotransmission of the mammalian central nervous system (CNS). They have physiological importance and clinical relevance in several diseases. We report the identification, cloning, and fine mapping of the human cDNA for GABAB receptor. A 4.2-Kb cDNA containing an open reading frame for a predicted protein of 960 aa was isolated from a fetal brain cDNA library. It had a strong identity (91.5%) with the rat GABAB receptor (rGB1A) nucleotide sequence, that corresponded to 98.6% identity at the amino acid level. Expression of the GABAB at the transcription level was detected by Northern analysis in all brain areas examined. The GABAB receptor has been mapped to human chromosome 6p21.3 within the HLA class I region close to the HLA-F gene. Susceptibility loci for multiple sclerosis, epilepsy, and schizophrenia have been suggested to map in this region. PMID- 9753615 TI - Direct evidence for peptide transporter (PepT1)-mediated uptake of a nonpeptide prodrug, valacyclovir. AB - Xenopus laevis oocytes were used as a gene expression system to characterize the carrier-mediated transport of valacyclovir (vacv), the L-valine ester prodrug of the acyclic nucleoside acyclovir (acv). A significant increase in the uptake of [3H]vacv by Xenopus laevis oocytes injected with human intestinal peptide transporter (hPepT1) cRNA compared to the uptake by water injected oocytes indicated that vacv was translocated by hPepT1. Vacv uptake was found to be concentration dependent, saturable (K(m) = 5.94 +/- 1.91 mM and Jmax = 1.68 +/- 0.25 nmoles/hr/oocyte), pH dependent, and inhibited by various known substrates of hPepT1 but not by acv, valine or pentaglycine. Vacv also inhibited the uptake of 14C-glycylsarcosine, a known substrate of hPepT1, in a concentration-dependent manner (Ki = 4.08 +/- 1.02 mM). These results demonstrate that human intestinal peptide transporter hPepT1 has broad specificity since it recognizes vacv as a substrate even though it lacks a typical peptide bond. PMID- 9753616 TI - Molecular cloning of a second human stanniocalcin homologue (STC2). AB - Stanniocalcin (STC) is a Ca- and phosphate-regulating hormone produced by the corpuscles of Stannius in bony fishes. The mammalian homologue of STC has recently been reported (STC1), which stimulates the phosphate uptake of kidney. Here we report the cloning of a second mammalian stanniocalcin (STC2) from the human osteosarcoma cDNA library. STC2 has 302 amino acid residues with 34% identity with STC1 and eel STC. STC2 has a conserved N-glycosylation site and is rich in cysteines as is the case with other stanniocalcins. STC2 has the same exon-intron boundaries as STC1. The culture medium of STC2-transfected CHO cells inhibited the promoter activity of Na-phosphate cotransporter (NaPi-3) and also inhibited the phosphate uptake of a kidney cell line (OK cells). Therefore, the function of STC2 seems to be opposite to that of STC1 on Na-phosphate cotransporter. Northern blot analysis revealed multiple transcripts in number of human tissues with high levels being present in skeletal muscle and heart. STC2 was also expressed in mice widely and its expression was lower in hypophosphatemic mice (Hyp mice) in many organs. We have cloned a widely expressed new human stanniocalcin homologue which suppressed the expression of renal Na-phosphate cotransporter. PMID- 9753617 TI - Reduced susceptibility of electroporated tumor cell lines to killing by cytotoxic lymphocytes. AB - Electroporation is a widely applied method for gene or protein transfer into cells, and it is also used for electrochemotherapy of cancer. During gene transfection studies, electroporation was found to decrease transiently susceptibility of some tumor cell lines to alloreactive cytotoxic T lymphocytes (CTL) or lymphokine-activated killer (LAK) cells. In each cell line electroporation induced c-fos mRNA. In K562 cells HSP70 mRNA induction also occurred. Expression of Grp78, Bcl-2, CD95/Fas, or major histocompatibility complex class I molecules was not affected by electroporation. PMID- 9753618 TI - A functional link between N-linked glycosylation and apoptosis in Chinese hamster ovary cells. AB - Seven different Chinese hamster ovary (CHO) cell mutants, isolated in different ways and having biochemical defects that were expressed at 34 degrees C, were found to be temperature sensitive for growth at 40.5 degrees C. Six of the mutants had five different lesions in N-linked glycosylation; two mutants were in the same complementation group. The temperature-sensitive phenotype in three mutants appeared by cell fusion studies to be linked to the glycosylation phenotype. In some of the glycosylation mutants [B4-2-1 (Lec15.1), Lec9, Lec1, and Lec24], but not in all of them (MI5-4 and MI8-5), incubation at 40.5 degrees C induced apoptosis, as determined by appearance of DNA fragmentation. Tunicamycin (TM) also induced apoptosis in both parental and Lec9 cells. There was a direct correlation between inhibition of glycosylation by TM treatment and induction of apoptosis. Induction of apoptosis by TM was inhibited by cycloheximide. These studies suggest that specific alterations in N-linked glycosylation in CHO cells are endogenous inducers of apoptosis. PMID- 9753619 TI - Increased insulin secretion and glucose tolerance in mice lacking islet amyloid polypeptide (amylin). AB - Islet amyloid polypeptide (IAPP or amylin) is costored and cosecreted with insulin and may regulate insulin secretion and blood glucose handling. However, the role and importance of endogenous IAPP in the regulation of insulin release and glucose homeostasis have been controversial. Here we report on the generation and phenotypic analysis of IAPP-deficient mice. These mice have normal, or near to normal, basal levels of circulating insulin and glucose. However, following glucose administration, IAPP-deficient males presented increased insulin responses paralleled with a more rapid blood glucose elimination compared to wild type controls. Blood glucose elimination was also found to be enhanced in IAPP deficient females, but the insulin response in this gender did not differ from controls. In a transgenic rescue experiment, using an insulin-promoter human-IAPP fusion gene, insulin responses and blood glucose elimination were reversed in IAPP-deficient males, whereas the female phenotype appeared unaffected. Our results provide the first firm evidence of a physiological role for endogenous IAPP and indicate that IAPP, apparently in a gender-dependent manner, limits the degree of glucose-induced insulin secretion and the rate of blood glucose elimination. PMID- 9753620 TI - Overexpression of glia maturation factor (GMF) in PC12 pheochromocytoma cells activates p38 MAP kinase, MAPKAP kinase-2, and tyrosine hydroxylase. AB - In order to study the intracellular regulatory function of glia maturation factor (GMF) in neuronal cells, we achieved a 10-fold overexpression of GMF in the rat pheochromocytoma cell line PC12 by infection with a replication-defective human adenovirus carrying a rat GMF cDNA insert. These cells showed a 3.6-fold increase in the activity of p38 MAP kinase, a 3.8-fold increase in the activity of MAPKAP K2 (MAP kinase-activated protein kinase 2), and a 4.2-fold increase in the activity of tyrosine hydroxylase (TH). We also detected an increase in the phosphorylation of TH and the 25-kDa heat shock protein (Hsp25) without a concomitant increase in their corresponding protein levels, suggesting a posttranslational modification. It was previously established that in PC12 cells, MAPKAP-K2 is an immediate target of p38, and both TH and Hsp25 are immediate targets of MAPKAP-K2. The current in vivo results are in concordance with our earlier in vitro finding that GMF promotes the activity of p38, and they implicate the participation of GMF in stress-induced catecholamine synthesis. PMID- 9753621 TI - Studies on the interaction between the vitamin D receptor and the radiolabelled 20-epi vitamin D analogue GS 1500. AB - Previous studies have shown that the binding affinity of a vitamin D analogue for the vitamin D receptor (VDR) does not correlate with the biological potency of the compound. In the present investigation the vitamin D analogue GS 1500, which is characterised by an altered stereochemistry at carbon C-20 (20-epi) and an aromatic ring in the side chain, was studied with respect to its interaction with the VDR. Using [3H]-GS 1500 as tracer, the receptor binding properties of GS 1500 were investigated and compared to those of 1,25(OH)2D3. The binding studies did not reveal a different binding site for GS 1500 than the one already established, and the binding affinity was in accordance with previously found values. At the level of VDR interaction with the vitamin D responsive element, GS 1500 did induce a binding complex at a lower concentration than 1,25(OH)2D3, which may help explain the difference in potency. PMID- 9753622 TI - Detection of mouse osteopontin by western blotting. AB - Several different antibodies to mouse and/or rat osteopontin have been developed, and some antisera raised against human osteopontin have been shown to react with mouse osteopontin. We have taken advantage of the lack of osteopontin protein in mice with a targeted disruption of the osteopontin gene to characterize the reactivity and the specificity of several of these antibodies with mouse osteopontin by Western blotting. Our results demonstrate that, with the exception of the rat monoclonal antibody MPIIIB10, which does not recognize mouse osteopontin on Western blots, all the tested reagents do react with mouse osteopontin, but their sensitivity varies widely, and in some cases there is significant cross-reactivity of the antibodies with other proteins found in mouse tissue extracts. PMID- 9753623 TI - Inhibition of proteasome function prevents thymocyte apoptosis: involvement of ornithine decarboxylase. AB - We have previously shown that polyamine levels rapidly decrease in thymocytes undergoing apoptosis, and that ornithine decarboxylase increases early but too transiently to maintain elevated polyamine levels. These data led us to suppose that a precocious ornithine decarboxylase degradation might be responsible for the imbalance of polyamine metabolism. Ornithine decarboxylase is known to be degraded by the cytosolic 26S proteasome that plays an essential role in thymocyte apoptosis. In this paper we demonstrate that the inhibition of proteasome function preserves ornithine decarboxylase activity and prevents thymocytes from undergoing apoptosis after dexamethasone treatment. Since intracellular polyamine levels are also preserved, ornithine decarboxylase seems to be functionally active in maintaining polyamine homeostasis after proteasome inhibition in thymocytes. Our proposed role for the proteasome in quiescent cells upon an apoptotic stimulus is to degrade proteins like ornithine decarboxylase that are involved in the control of the cell cycle and cell survival. PMID- 9753624 TI - Urocortin, a newly identified corticotropin-releasing factor-related mammalian peptide, stimulates atrial natriuretic peptide and brain natriuretic peptide secretions from neonatal rat cardiomyocytes. AB - The effect of urocortin (UCN), a recently characterized mammalian member of corticotropin-releasing factor (CRF)-related peptide and a putative endogenous ligand for CRF type 2 beta receptor in the regulation of atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) release, was investigated using cultured neonatal rat cardiomyocytes. Treatment with UCN (10(-10)-10(-6)M) resulted in significant increase in ANP and BNP secretions, and the effect of UCN on ANP and BNP secretions was more potent than that of CRF on an equimolar basis. The effect of UCN (10(-7)M) was completely blocked by alpha-helical CRF (9-41), a specific CRF type 2 receptor antagonist. The effect of UCN (10(-7)M) was not only blunted by cAMP-dependent protein kinase A (PKA) inhibitor, H-89 (10(-5)M), but also diltiazem (10(-7)M), a voltage-dependent Ca2+ channel blocker. Further, UCN stimulated cAMP production in cardiomyocytes. Also, UCN (10(-7)M) itself stimulated [3H]leucine uptake into neonatal rat cardiomyocytes and potentiated endothelin-1-induced increase of [3H]leucine uptake. These results suggest that activation of CRF type 2 receptor, especially type 2 beta receptor, with UCN induces ANP and BNP secretions, at least in part, via PKA pathway during cardiac hypertrophy. PMID- 9753625 TI - Human CART1, a paired-class homeodomain protein, activates transcription through palindromic binding sites. AB - Homeodomain proteins play important roles in animal development by controlling the expression of genes involved in determining cell fates. The recently cloned human Cart1 gene encodes a paired-class homeodomain (hCART1), whose rodent homolog is mainly expressed in early chondrocytes and in prechondrocytic mesenchymal cells. To better understand its role as a transcription factor, the author has selected specific hCART1 binding sites from a random pool of oligonucleotides. It is reported here that all sites obtained contain a palindrome consisting of two TAAT sequences separated by three or four base pairs. In electromobility shift assays, recombinant hCART1 proteins bind to a palindromic probe as a multimer, possibly a homodimer. In transient transfection assays, hCART1 activates transcription from reporter plasmids containing hCART1 binding sites in HeLa cells, demonstrating both site-dependence and dosage dependence. It is also shown here that hCART1 localizes to nucleus. These data indicate that hCART1 is a sequence-specific transcription activator in HeLa cells. In combination with data from previous studies in which hCART1 represses transcription in different cell types and promoters, they suggest that hCART1 may be a transcription modulator with both repression and activation activities. PMID- 9753626 TI - At least two receptors of asymmetric acetylcholinesterase are present at the synaptic basal lamina of Torpedo electric organ. AB - Asymmetric acetylcholinesterase (AChE) is anchored to the basal lamina (BL) of cholinergic synapses via its collagenic tail, yet the complement of matrix receptors involved in its attachment remains unknown. The development of a novel overlay technique has allowed us to identify two Torpedo BL components that bind asymmetric AChE: a polypeptide of approximately 140 kDa and a doublet of 195-215 kDa. These were found to stain metachromatically with Coomassie blue R-250, were solubilized by acetic acid, and were sensitive to collagenase treatment. Upon sequence analysis, the 140 kDa polypeptide yielded a characteristic collagenous motif. Another AChE-binding BL constituent, identified by overlay, corresponded to a heparan sulfate proteoglycan. Lastly, we established that this proteoglycan, but not the collagenous proteins, interacted with at least one heparin binding domain of the collagenic tail of AChE. Our results indicate that at least two BL receptors are likely to exist for asymmetric AChE in Torpedo electric organ. PMID- 9753627 TI - Function of steroidogenic factor 1 (SF1) ligand-binding domain in gene activation and interaction with AP1. AB - The nuclear receptor SF1 is an essential mediator in ventromedial hypothalamus pituitary-gonadal development. As with other nuclear receptors, SF1 possesses a DNA-binding domain composed of two zinc fingers and a ligand-binding domain containing a ligand-dependent activation sequence termed AF2. To dissect the domain function of SF1, we examined various SF1 mutants in mouse adrenocortical Y1 cells and human placental JEG3 cells. Destruction of the AF2 structure removed 73-90% transactivation activity, suggesting that AF2 is indispensable for transactivation. Mutants carrying the DNA-binding domain but lacking the AF2 or the ligand-binding domain blocked the activity of normal SF1. Disrupting the zinc finger diminished the dominant negative effect of mutant. Cotransfection of SF1 with AP1 showed that the two transcription factors cooperated to activate gene expression. Some mutants lost the synergistic action with AP1, while some retained partial activity. These experiments delineate the functional domains of SF1. PMID- 9753628 TI - Normal myoblast implantation in MDX mice prevents muscle damage by exercise. AB - One consequence of the lack of dystrophin is a higher vulnerability of myofibers to eccentric exercise. In this study, we compared the effect of downhill running on Biceps brachii of MDX mice with or without transplantation of normal myoblasts. Exercise induced damaged was detected by Evans blue staining. In control MDX mice, 26.3% of the fibers were permeated by this dye, myoblast transplantation prevented such necrosis. In the transplanted muscles, only dystrophin negative fibers were injured. Indeed, in muscles containing at least 40% dystrophin positive fibers, the damage was significantly reduced in the grafted muscle. Thus the transplantation of normal myoblasts increases the resistance of dystrophic muscles to exercise. Our results suggest that transplantation of normal myoblasts to DMD patients may have beneficial effects. PMID- 9753629 TI - CFTR regions containing duodenum specific DNase I hypersensitive sites drive expression in intestinal crypt cells but not in fibroblasts. AB - We have investigated CFTR specific intestinal expression by transfection assays in mouse cultured fibroblasts and transimmortalized intestinal crypt m-ICc12 cells using the beta-galactosidase gene linked to rat CFTR non-coding regions. Two constructs were studied, one encompassing a 5.3 kb region 5' to the gene where numerous duodenum-specific DNase I hypersensitive sites (DHSs) were previously mapped and the other including a 1.3 kb 3' region in which novel DHSs had been identified. In transient transfection assays, transgenes were expressed in m-ICc12 cells but not in fibroblasts. In m-ICc12 cells, the pattern of expression of the chromosomally integrated transgenes paralleled the endogenous expression of CFTR and beta-galactosidase activity was detected in cells containing villin and forming domes. Thus, a 6.6 kb region encompassing 5' and 3' non-coding parts of rat CFTR is able to drive specific expression of a reporter gene in cultured mouse intestinal cells having kept a crypt phenotype. PMID- 9753630 TI - The subcellular location of the yeast Saccharomyces cerevisiae homologue of the protein defective in the juvenile form of Batten disease. AB - The mutation responsible for the juvenile form of Batten disease was mapped to a single gene, Cln3 (T. J. Lerner et al. (1995) Cell 82:949-957). Yeast Saccharomyces cerevisiae has an open reading frame, BTN1 (YHC3), that encodes the putative homologue of Cln3p. Primary structure comparison indicates that the human Cln3p and yeast Btn1p are 59% similar and 39% identical and they have similar hydropathy profiles. Gene disruption of BTN1 in yeast has no apparent effect on growth or viability of the cells under a variety of conditions. Gene fusion protein constructs of green fluorescent protein (GFP) and Btn1p, with GFP at the amino and carboxyl ends of Btn1p, localize to the vacuole in yeast. These data indicate that BTN1 is a nonessential gene under most growth conditions which functions in the vacuole in yeast Saccharomyces cerevisiae. PMID- 9753631 TI - Cooperativity of mutational effects within a six amino acid residues substitution that induces a major conformational change in human H ferritin. AB - Ferritin is an iron-storage protein composed of 24 polypeptide chains which assemble into a hollow shell. Previously, we have shown that a multisubstituted ferritin mutant containing the peptide ESVWNP in place of the wild-type sequence GAPESG in a short exposed loop directs the synthesis of a product that assembles in a conformation remarkably different from that of the normal molecule. We have further characterized this mutant and we have tried to determine which of the substituted residues causes the large conformational change. Reversion of the mutant conformation was obtained changing the three residues WNP back to the wild type sequence ESG (DE loop: ESVESG). However, the converse three amino acid change GAPWNP produced insoluble and unassembled ferritin. Therefore, the substitutions of GAP by ESV together with ESG by WNP have a largely cooperative and hardly predictable effect. PMID- 9753632 TI - Two proteins translated by alternative usage of initiation codons in mRNA encoding a JunD transcriptional regulator. AB - The junD gene encodes one component of the transcription factor, AP-1. Since two forms of JunD protein have been reported, we analyzed here the molecular mechanisms involved in the isoform production. Immunochemical analysis indicated that the longer and shorter forms of mouse JunD (JunD-L and JunD-S, with apparent molecular weights of 44 and 39 kDa, respectively) differ in their content of an N terminal peptide. Mutational analysis further indicated that JunD-S is the translational product initiated at the third AUG located 144 bp from the first AUG, at which JunD-L translation starts. Such production of two junD isoforms from a single mRNA using the same reading frame seems to be conserved in human, rat, and chicken. To examine the functional differences between the isoforms, each type of JunD was exclusively expressed by the use of retrovirus vectors harboring the mutated junD gene. The exogenous expression of either one of these forms did not cause cellular transformation of NIH3T3, but suppressed the anchorage-independent growth of NIH3T3 transfor-bold by the activated K-ras or v src gene. These two isoforms were expressed in all the mouse tissues examined and in various cell lines established from human tumors, though the expression ratio between JunD-L and JunD-S varied, suggesting that some factor(s) modulate the alternative usage of the initiation codon of the junD gene. PMID- 9753633 TI - Autoxidation of pyridoxalated hemoglobin polyoxyethylene conjugate. AB - Hemoglobin-based therapeutics are currently in clinical trials in the United States and abroad as blood replacement solutions, nitric oxide scavengers, and radiation sensitizers. The potency of the therapeutics may be influenced by the oxidation state of the iron in the heme moiety. The oxidation state is dependent upon the physical environment of the molecule and is influenced by parameters such as the chemical nature of the hemoglobin therapeutic and its formulation. Pyridoxalated hemoglobin polyoxyethylene conjugate (PHP) is one such compound currently in clinical trials in the U.S. for treatment of nitric oxide-dependent, volume refractory shock. The autoxidation rates for PHP have been determined over a range of temperatures. The oxidation events were shown to be biphasic and were similar to those observed for purified human hemoglobin (HbAo). The initial fast oxidation events were modeled with first order rate constants at 37 degrees C and determined to be 0.022 hr-1 and 0.025 hr-1 for PHP and HbAo, respectively. The autoxidation of PHP was shown to be independent of concentration from approximately 5 to 100 mg/mL. PMID- 9753634 TI - Identification and characterization in Xenopus of XsmgGDS, a RalB binding protein. AB - We have previously isolated a cDNA coding for a RalB protein from a Xenopus maternal library. By mutagenesis of RalB and microinjection of its mRNA in embryos we show that RalB is involved in Xenopus early development. We have used the yeast two-hybrid system to screen a Xenopus maternal cDNA library in order to isolate proteins interacting to RalB. We have identified a full-length cDNA encoding a protein homologous to mammalian smgGDS. The Xenopus gene shows 84.6% identity with the mammalian counterpart and contains one additional armadillo repeat. The XsmgGDS mRNA is expressed from oogenesis up to late embryogenesis at a higher level than that in adult tissues. Thus RalB is another protein that interacts with smgGDS which suggests that RalB may be activated by a Ras independent pathway. PMID- 9753635 TI - Detection of a nitric oxide synthase possibly involved in the regulation of the Rhodococcus sp R312 nitrile hydratase. AB - Crude homogenates from Rhodococcus sp 312 catalyze the conversion of L-arginine into L-citrulline and NO2-, the usual oxidation product of NO under aerobic conditions. They also catalyze the conversion of N omega-hydroxy-L-arginine (NOHA) into L-citrulline and NO2- with similar rates (10-15 and 100-150 nmol of product.min-1.(mg of protein)-1 respectively for the crude homogenate and for a fraction obtained from ammonium sulfate precipitation). L-citrulline formation is strongly inhibited by classical inhibitors of mammalian nitric oxide synthases (NOSs) such as N omega-methyl-L-arginine (NMA) and thio-L-citrulline (TC). Finally, the lack of inhibitory effects of EGTA, a classical inhibitor of constitutive mammalian NOSs, and the specific immunodetection of a 100 kD protein from Rhodococcus cytosol by an antibody raised against human inducible NOS, is in favor of the presence of a NOS similar to inducible mammalian NOSs in Rhodococcus sp 312. This NOS should be responsible for the NO-dependent inactivation of Rhodococcus Nitrile Hydratase (NHase) in the absence of light; it could regulate the activity of the latter enzyme. PMID- 9753636 TI - Plasmalogen phospholipids are involved in HDL-mediated cholesterol efflux: insights from investigations with plasmalogen-deficient cells. AB - Plasmalogens are ether-glycerophospholipids that exist in all mammalian cells, but their physiological function remains thus far an enigma. It has been previously suggested that the association of high-density lipoprotein (HDL) with cellular phospholipid is a pre-requisite for the process of HDL-mediated cholesterol efflux (HDL-MCE). To investigate our hypothesis that plasmalogens might play a role in HDL-MCE, we used a model composed of plasmalogen-deficient cells including RAW mutant macrophages and fibroblasts from patients with rhizomelic chondrodysplasia punctata type II. In mutant macrophages, HDL-MCE was reduced by 57% compared to control macrophages, after 16 hours. A similar phenomenon was observed in plasmalogen-deficient patients fibroblasts. Incubation of plasmalogen-deficient fibroblasts with 1-0-hexadecyl-sn-glycerol, which restored plasmalogen levels to that of control cells, resulted in a 35% increase in HDL-MCE, compared to a 10% increment in controls. The novel finding that HDL MCE is reduced in plasmalogen-deficient cells and increases following plasmalogen restoration leads us to suggest that plasmalogen has an important function in the mediation of cellular cholesterol efflux. PMID- 9753637 TI - Senescence marker protein-30 (SMP30) rescues cell death by enhancing plasma membrane Ca(2+)-pumping activity in Hep G2 cells. AB - Senescence marker protein-30 (SMP30) has been reported to decrease with aging in the rat liver. SMP30 has been also suggested to play a role as a Ca(2+)-binding protein localized in cytosol of hepatocytes. To elucidate the functional significance of SMP30, we have generated Hep G2 cell lines that stably express large amounts of SMP30 by transfection with human SMP30 cDNA. Using these cell lines, in view of the intracellular Ca2+ homeostasis, we then investigated cytosolic free Ca2+ concentration ([Ca2+]i) and Na(+)-independent Ca2+ efflux from the cells after extracellular ATP stimulation. Although stimulation of cells with ATP caused transient [Ca2+]i increase in both SMP30 and mock transfectants, rate of decrease after peak in [Ca2+]i was enhanced 2-fold by transfection of SMP30. Correspondingly, Ca2+ efflux was significantly increased in SMP30 transfectants compared with mock transfectants. In addition, more SMP30 transfectants survived than mock transfectants when cell death was induced by Ca2+ ionophore treatment. These results suggest that SMP30 regulates [Ca2+]i by modulating plasma membrane Ca(2+)-pumping activity, and thus down-regulation of SMP30 during aging may contribute to deterioration of cellular functions. PMID- 9753638 TI - Autophosphorylation of enzyme I of the Escherichia coli phosphoenolpyruvate:sugar phosphotransferase system requires dimerization. AB - Enzyme I of the Escherichia coli phosphoenolpyruvate:sugar phosphotransferase system undergoes a slow monomer-dimer transition. In vitro autophosphorylation of Enzyme I by PEP was studied at limiting concentrations of the protein. Addition to incubation mixtures containing wild-type Enzyme I of inactive or low-activity mutant forms of Enzyme I resulted in stimulation of autophosphorylation activity. The kinetics of the activation fit well to a model in which the active form of Enzyme I is the dimer. These experiments provide support for the argument that only the dimeric form of Enzyme I can be autophosphorylated. PMID- 9753639 TI - Transition metals bind to glycated proteins forming redox active "glycochelates": implications for the pathogenesis of certain diabetic complications. AB - The present investigations arose from our interest in the possibility that some structures which arise secondary to protein glycation might bind transition metals such as iron and copper. In support of this we find that, when glycated, three different proteins--albumin, gelatin (a soluble collagen fragment) and elastin--all gain a substantial affinity for the transition metals iron and copper. The glycated proteins bind at least three times as much iron as do the non-glycated proteins. Similarly, glycated albumin and gelatin also bind 2-3 times as much copper. Furthermore, at least in the case of copper bound to glycated albumin, the bound metal retains redox activity and participates in the catalytic oxidation of ascorbic acid. Should similar "glycochelates" form in vivo in diabetics, reactions mediated by these chelates may be involved in certain complications of diabetes. PMID- 9753640 TI - Growth inhibition of HL-60 cells by extracellular ATP: concentration-dependent involvement of a P2 receptor and adenosine generation. AB - A single addition of ATP (20-1000 microM) to cultures of HL-60 cells resulted here in permanent, Ca(2+)-independent inhibition of cellular proliferation, evident 48 h following treatment. Extracellular ATP (ATPo) was maximally effective at 250 microM giving 90 +/- 1.5% growth inhibition. Up to a concentration of 250 microM ATPo, growth inhibition is solely attributable to ATPo, while at higher ATPo concentrations adenosine generated from ATPo hydrolysis contributes to this effect. The order of potency for growth inhibition was ATP = ADP > AMP > adenosine. Suramin, a P2 receptor antagonist, attenuated growth inhibition by ATP and ADP, indicative of P2 receptor involvement. Equipotency of ATP and ADP excludes the involvement of either an ecto-protein kinase or a P2X7 receptor in growth inhibition. Neither UTP (P2Y2 agonist) nor alpha, beta-methyleneATP (P2X1 agonist) inhibited growth, indicating that such inhibition is mediated by a previously undescribed P2 receptor on HL-60 cells. PMID- 9753641 TI - Characterization of the C-terminal domain of Helicobacter pylori vacuolating toxin and its relationship with extracellular toxin production. AB - Helicobacter pylori vacuolating cytotoxin (VacA) induces gastric epithelial necrosis. Its C-terminal domain is hypothesized to be responsible for extracellular translocation of the mature cytotoxin. In this study, genetic structural properties of VacA C-terminal domain and the level of cytotoxin secretion were investigated. Sau3AI-HaeIII restriction fragment length polymorphism (RFLP) analysis of the 1.1-kb PCR-amplified vacA fragment revealed 14 distinct combined patterns among 87 clinical isolates. Of the 4 popular groups (A-a, A-b, A-f, and B-a), A-a strains produced a higher level of the VacA protein than A-b strains and than A-f strains (P < 0.05). Sequence analysis and secondary structure prediction supported a beta-barrel structure that might act as a selective export channel like Iga beta-core of IgA proteases. Sequence differences in the predicted beta-barrel were present among strains of different RFLPs. PMID- 9753642 TI - Use of fluorescein labelled antibody and fluorescence activated cell sorter for rapid identification of Mycobacterium species. AB - A fluorescein labelled antibody (Ab)/Fluorescence Activated Cell Sorter (FACS) based assay was developed for detection of a wide range of mycobacterial species directly from bacterial culture and sputum specimens. The whole process could be completed within 3 hours and had a high specificity and sensitivity for cultured bacteria. The method was also shown to be applicable for direct identification from clinical specimens. This study showed that pretesting of clinical specimens for mycobacteria to the genus level with an antibody to Mycobacterium species offers the routine clinical laboratory a single convenient test for the detection of tuberculous and nontuberculous mycobacteria. Depending on the availability of species-specific antibody, the identification of Mycobacterium to the species level can be achieved. PMID- 9753643 TI - Gene synthesis, expression, and mutagenesis of zucchini mavicyanin: the fourth ligand of blue copper center is Gln. AB - The gene coding for the 109-amino-acid, non-glycosylated form of mavicyanin was synthesized and expressed in Escherichia coli. The recombinant protein refolded from E. coli inclusion bodies was purified and characterized. Its spectroscopic properties are fully identical to those of mavicyanin isolated from zucchini, even in the absence of its carbohydrate moiety. The blue cooper center of mavicyanin strongly binds three ligands (2His and Cys) as well as many blue copper proteins. To disclose the fourth ligand of mavicyanin, Met was substituted for Gln95 by site-directed mutagenesis. The replacement changes from a rhombic EPR signal to an axial one and exhibits the quite similar absorption and CD spectra to those of plastocyanin. The midpoint potential of Gln95-->Met mavicyanin shows the positive shift of 187 mV compared with the recombinant protein, being close to the values of plastocyanins. The differences of the spectroscopic and electrochemical properties between mavicyanin and its mutant demonstrate that the fourth ligand of mavicyanin is Gln95. PMID- 9753644 TI - Heat-shock protein 90 (hsp90) binds in vitro to tubulin dimer and inhibits microtubule formation. AB - Hsp90 interacts with steroid hormone receptors, protein kinases, and cytoskeletal proteins. The mode of action of hsp90 on microtubules and tubulin has not been investigated. Using isolated purified hsp90 and isolated tubulin, we demonstrated in vitro by difference absorption and fluorescence spectroscopy that hsp90 bound to tubulin with an apparent affinity constant of 5 x 10(5) M-1, assuming an apparent stoichiometry of 1 at 25 degrees C. Using microcalorimetry, we found a delta H of -9.8 +/- 0.8 kJ.mol-1. The binding of hsp90 to tubulin was confirmed by a sedimentation assay. Moreover, we showed that hsp90 inhibited tubulin polymerisation. PMID- 9753645 TI - Analysis of temporal expression pattern and cis-regulatory sequences of chicken pepsinogen A and C. AB - Three groups of pepsinogens exist in vertebrates, namely, pepsinogen A, pepsinogen C, and prochymosin, which are produced at different developmental stages. In the chicken, prochymosin is expressed only in the embryonic stage, while pepsinogens A and C are secreted from adult chicken proventricular (glandular stomach) mucosa. In order to understand the mechanism of transcriptional regulation of these genes, we have cloned the genes encoding chicken pepsinogens A and C and analyzed the sequences possibly involved in their regulation. 5'-Upstream sequences of both genes contain putative binding motifs for transcription factors such as GATA, Sox, and HNF-3 beta, which are expressed in the chicken gut epithelium. Moreover, we found seven putative binding motifs for human MZF-1 in intron 8 of pepsinogen A gene. These transcription factors may act as regulators of expression of chicken pepsinogen genes. PMID- 9753646 TI - Protein A induces NO production: involvement of tyrosine kinase, phospholipase C, and protein kinase C. AB - Protein A of S. aureus exhibits a wide array of immunopotentiating activities. Since the role of nitric oxide (NO) in bioregulation has been well envisaged; we studied the effect of Protein A on NO production by immunocytes both in vivo and in vitro. Our data indicate that PA at a comparable dose of LPS (lipopolysaccharide) increases the NO levels in the serum of Swiss albino mice by about 12-fold from its basal level. The peak level is reached at about 12 hours after i.p. inoculation of PA. However, NO concentration returns to the basal value 15 hours posttreatment. Splenic lymphocytes and peritoneal macrophages showed appreciable increase in NO production when cultured with PA in vitro. Interestingly, inhibitors of tyrosine kinase, phospholipase C, and protein kinase C (PKC) inhibited NO production in splenic lymphocytes. Thus, it appears that these enzymes participate in the signaling cascade induced by PA, which culminates in the production of NO downstream of PKC. It is possible that PA induced NO production may have relevance with the anti-tumor and anti-parasitic properties of PA, described earlier. PMID- 9753647 TI - Involvement of MAP kinase-independent protein kinase C signaling pathway in the EGF-induced p21(WAF1/Cip1) expression and growth inhibition of A431 cells. AB - Previous studies have revealed that the growth inhibition of A431 cells overexpressing epidermal growth factor (EGF) receptors by a high concentration of EGF is mainly due to the expression of cycline dependent kinase (CDK) inhibitor p21(WAF1/Cip1). However, the signal transduction mechanism from the activated EGF receptor to the induction of p21(WAF1/Cip1) gene is still poorly understood. We investigated which signaling pathway plays an important role in p21(WAF1/Cip1) expression and growth inhibition by using specific inhibitors of the signaling molecules. A broad PKC inhibitor, PKC delta inhibitor, but not the conventional PKC inhibitor suppressed the EGF-induced p21(WAF1/Cip1) expression and the growth inhibition of A431 cells. These inhibitors did not alter either the activation of EGF receptor or the stimulation of MAP kinase at detectable levels. Furthermore, we found that the induction of p21(WAF1/Cip1) at the early phase (within 12 hr after stimulation) by a high concentration of EGF was independent of the MAP kinase activation by using dominant negative Ras. These results suggest that PKC, especially PKC delta plays a crucial role in the EGF-induced p21(WAF1/Cip1) expression, resulting in the growth inhibition of A431 cells. PMID- 9753648 TI - Reconstitution of IL6-inducible differentiation of a myeloid leukemia cell line by activated Stat factors. AB - Differentiation of the myeloid leukemia cell line M1 by treatment with IL6-type cytokines depends on activation of the Jak/Stat (Janus kinase/signal transducer and activator of transcription) pathway. Defects in this cascade are correlated with an impaired cytokine-inducible differentiation of various other myeloid cell lines. Although treatment with IL-6 increased the amount of activated transcription factor Stat3 in the myeloid leukemia line C, differentiation was not induced. However, after cotransfection with expression constructs for the tyrosine kinase Jak2 and Stat factors 3 or 5a, treatment of the cells with IL-6 caused a decrease in the number of viable cells. In parallel, an increase in the percentage of differentiated cells occurred. These findings are consistent with the hypothesis that the Jak/Stat signaling cascade plays an important role in cytokine-induced differentiation of myeloid leukemia cells. PMID- 9753649 TI - Novel biological functions of interleukin-4: formation of tube-like structures by vascular endothelial cells in vitro and angiogenesis in vivo. AB - The effect of interleukin-4 (IL-4) on angiogenesis was studied in vitro and in vivo. Human recombinant IL-4 significantly stimulated the formation of tube-like structures in collagen gels by bovine aortic endothelial cells as well as by human microvascular endothelial cells. Human recombinant IL-4 at 50-500 U/ml stimulated by about two- to threefold the formation of tubes by bovine aortic endothelial cells; the rate was comparable to that of basic fibroblast growth factor. Tube formation was almost completely inhibited by the addition of IL-4 receptor neutralizing antibody. Administration of rat recombinant IL-4 led to neovascularization when implanted in the rat cornea. Findings suggest that IL-4 may be a mediator of the immune system as well as an inducer of angiogenesis. PMID- 9753650 TI - Increased cell surface exposure of fucose residues is a late event in apoptosis. AB - Apoptosis is a well defined physiological process characterised by many specific features including DNA fragmentation and protease activation. Cell membrane associated changes such as the altered exposure of phosphatidylserine and glycosylation patterns have also been described. We investigate here the change in exposure of surface alpha-L-fucose residues during thymocyte and P815 cell apoptosis. We show that apoptosis in these cells induced by dexamethasone, gliotoxin or thapsigargin was associated with an increase in the exposure of terminal fucose residues. Furthermore, this increase in fucose exposure occurred late in the apoptotic process. The observation of increased fucose exposure in two different cell types by three different apoptosis-inducing agents suggests it may be part of the normal apoptotic process. PMID- 9753651 TI - dATP causes specific release of cytochrome C from mitochondria. AB - The induction of the Mitochondrial Permeability Transition (MPT) has recently been associated with the release of apoptogenic cytochrome c, which could come about in a swelling-dependent or swelling-independent manner. We observed that canonical inducers of MPT (Ca2+, t-butyl hydroperoxide, atractyloside) induce a swelling-dependent release of cytochrome c, and that osmotic support of mitochondria with PEG-1000 abolishes mitochondrial swelling, protein release, and cytochrome c release by these inducers. By contrast, it was observed that dATP is a potent inducer that caused release of cytochrome c in a swelling independent manner, i.e. even in the presence of osmotic support by PEG-1000; in addition this release of cytochrome c is inhibitable by cyclosporin A. The dATP-dependent and swelling-independent release of cytochrome c from mitochondria is not inhibitable by the protease inhibitor z-VAD, suggesting that it is not mediated by upstream caspases. This is the first report to our knowledge that a chemical compound may directly cause release of cytochrome c from mitochondria, and could explain the toxicity of dATP in the context of the genetic immunodeficiency diseases Adenosine Deaminase deficiency and Purine Nucleotide Phosphorylase deficiency. PMID- 9753652 TI - Pulsed electromagnetic fields simultaneously induce osteogenesis and upregulate transcription of bone morphogenetic proteins 2 and 4 in rat osteoblasts in vitro. AB - Pulsed electromagnetic fields (PEMF) are successfully employed in the treatment of a variety of orthopaedic conditions, particularly delayed and nonunion fractures. In this study, we examined PEMF effects on in vitro osteogenesis by bone nodule formation and on mRNA expression of bone morphogenetic proteins 2 and 4 by reverse-transcriptase polymerase chain reaction (RT-PCR) in cultured rat calvarial osteoblasts. PEMF exposure induced a significant increase in both the number (39% over unexposed controls) and size (70% larger compared to unexposed controls) of bone-like nodules formed. PEMF also induced an increase in the levels of BMP-2 and BMP-4 mRNA in comparison to controls. This effect was directly related to the duration of PEMF exposure. This study shows that clinically applied PEMF have a reproducible osteogenic effect in vitro and simultaneously induce BMP-2 and -4 mRNA transcription. This supports the concept that the two effects are related. PMID- 9753653 TI - Identification and analysis of the regulatory region R&R* with the cnf1 gene encoding the cytotoxic necrotizing factor type 1 that closely links to the lux regulon of Vibrio fischeri. AB - Nucleotide sequence of the regulatory region R&R* and the partial 5'-end of the cnf1 gene (GenBank Accession No. AF023157) of Vibrio fischeri ATCC 7744 has been determined, and the cytotoxic necrotizing factor 1 (CNF1) encoded by the cnf1 gene is deduced. Alignment and comparison of the cytotoxic necrotizing factor 1s of V. fischeri and E. coli show that they are homologous. Nucleotide sequence reveals that the cnf1 gene is closely linked to the lux regulon in genome; the gene order of the cnf1 gene and the lux regulon is <--cnf1-R&R*<--rrn-<--luxR-R&R luxI-luxC-luxD -luxA-luxB-luxE-luxG-omega-->, whereas R&R is the regulatory region of the lux regulon, and R&R* is the regulatory region of the cnf1 gene; the sequence approximately 2 kb lay between the luxR gene of the lux regulon and the cnf1 gene is an rrn-like operon. It is unexpected to find the cnf1 gene in V. fischeri, since the CNF1 protein enables necrosis; the marine luminous bacterium V. fischeri is never to be identified as a pathogenic microbe. The cnf1 gene might be concerned with symbosis of the luminous bacteria and host fishes. PMID- 9753654 TI - An essential methionine residue involved in substrate binding by phosphofructokinases. AB - An alignment of all PPi-dependent phosphofructokinases and all allosteric ATP dependent PFKs shows relatively few residues that are fully conserved. One residue that is conserved is a methionine residue that appears from the crystal structure of Escherichia coli PFK to be interacting with fructose 6-P. Very conservative substitutions for this methionine with leucine or isoleucine by site directed mutagenesis of E. coli ATP-PFK and Entamoeba histolytica PPi-PFK produced profound decreases either in the apparent affinity for fructose 6-P or in maximal velocity, or both. Methionine provides a highly specific interaction with fructose 6-P for binding and for transition state stabilization. PMID- 9753655 TI - Serines at the active site of 11 beta-hydroxysteroid dehydrogenase type I determine the rate of catalysis. AB - Short chain alcohol dehydrogenases have an invariant YXXXK motif at the active site. Database analysis of 116 superfamily members showed that 92% also contain a Serine or Threonine residue at the Y + 1 or Y + 3 positions, a pattern we previously described as the ST rule. In the present study we have mutated Serines in the active site, YSASK, motif of 11 beta-hydroxysteroid dehydrogenase type I (11 beta HSD1). These studies were facilitated by the generation of a new specific polyclonal antibody (RAH113) raised against a synthetic peptide derived from the rat 11 beta-HSD1 sequence. Immunopurified RAH113 recognized a single band at 34 kDa in liver homogenates. Kinetic analysis of equivalent amounts of wild type and mutant proteins showed that mutagenesis of active site Serines resulted in modest increases of Km values for corticosterone from 325 nM for 11 beta HSD1 to 512 nM-588 nM for the S1 (YAASK), S2 (YSAGK) and S3 (YAAGK) mutants in homogenates of transfected CHOP cells. However, far greater effects were observed on the first order rate constants with mutants displaying 10%, 1% and 1% of the wild type activity, respectively. When the oxidoreductase reaction was studied in whole cells mutagenesis again had a minimal effect on the Km value but dramatically lowered first order rate constants to 34%, 5% and 6%, respectively, of the wild type. These data show that Serines at the active site of 11 beta HSD1 play an important role in determining the rate of catalysis. Coexpression of wild type and mutant enzymes did not lower wild type activity suggesting that the active site of the multimeric enzyme is not a composite of active site Serines from different subunits. PMID- 9753656 TI - Histatin 5 is a substrate and not an inhibitor of the Arg- and Lys-specific proteinases of Porphyromonas gingivalis. AB - The salivary peptide histatin 5 has been reported to be an inhibitor of the Arg- and Lys-specific proteinases of Porphyromonas gingivalis, an oral pathogen associated with periodontitis. In this study a purified P. gingivalis proteinase preparation consisting of a complex of the Arg- and Lys-specific proteinases and adhesins was assayed using chromogenic substrates in the presence of histatin 5. Histatin 5 produced a concentration-dependent decrease in the initial rate of hydrolysis of the chromogenic substrates by both proteinases. However, pre incubation of histatin 5 with the purified proteinase preparation or a P. gingivalis cell sonicate for 10 min prior to assay with the chromogenic substrates showed that under these conditions the salivary peptide did not decrease the initial rate of chromogen release. Mass spectrometric analysis revealed rapid degradation of histatin 5 at all four lysyl and all three arginyl residues by the P. gingivalis proteinases. This study demonstrates that histatin 5 is a substrate for the P. gingivalis extracellular Arg- and Lys-specific cysteine proteinases and not an inhibitor. PMID- 9753657 TI - Plasmatocyte spreading peptide is encoded by an mRNA differentially expressed in tissues of the moth Pseudoplusia includens. AB - It has long been known that blood cells (hemocytes) are an essential component of the invertebrate immune system, yet little is known about the molecules mediating their function. Recently, we identified plasmatocyte spreading peptide (PSP1) from the moth Pseudoplusia includens which regulates the trafficking and adhesion of a hemocyte subclass called plasmatocytes. Here, we report the cloning of a cDNA (p15) that encodes a PSP1 precursor protein. Northern blot analysis revealed that a homologous prepro-PSP1 mRNA is expressed in fat body, and that other PSP1 related transcripts are expressed in nervous tissue and hemocytes. Coupled in vitro transcription/translation reactions indicated that p15 produces a protein recognized by a PSP1 polyclonal antibody. Immunoblotting experiments further revealed that a putative pro-PSP1 protein is present in P. includens plasma and fat body. PMID- 9753658 TI - A novel hepatic stellate (Ito) cell-derived protein, epimorphin, plays a key role in the late stages of liver regeneration. AB - Limited data exist regarding morphogenesis and differentiation during liver regeneration. We examined the role of epimorphin on liver regeneration. After 70% partial hepatectomy, mouse liver was collected on days 1, 3, 7, and 14 for immunohistochemistry and the detection of epimorphin mRNA and connexin 32. Using primary cultured rat hepatocytes, morphogenesis and differentiation of cells were tested with or without epimorphin. Seven days after cell inoculation, the expression of connexin 32 and the cell-cell communication was tested as a marker of differentiation. Epimorphin was detected exclusively in hepatic stellate cells. Connexin 32 was detected only in hepatocytes. After partial hepatectomy, epimorphin mRNA was detected on day 3 and peaked at day 7, followed by protein expression. Connexin 32 expression showed a similar time course. Cultured hepatocytes formed multicellular spheroids in an active epimorphin-coated culture dish and showed positive dye coupling, whereas the cell-cell communication was lost without active epimorphin. Because epimorphin was expressed late in liver regeneration, it might play a role in morphogenesis and differentiation. PMID- 9753659 TI - H-ras activation is an early event in the ptaquiloside-induced carcinogenesis: comparison of acute and chronic toxicity in rats. AB - Bracken fern (Pteridium spp.) produces cancer of the urinary bladder and oesophagus in grazing animals and is a suspected human carcinogen. The carcinogenic principle ptaquiloside (PT), when activated to a dienone (APT), forms DNA adducts which eventually leads to tumor. Two groups of female Sprague Dawley rats were given a chronic dose of 3 mg APT weekly for 10 weeks either by intravenous (i.v.) tail vein or by intragastric (i.g.) route. A third group was given a weekly dose of 6 mg of APT for 3 weeks by the i.g. route corresponding to acute dosing. Both chronic i.v. and i.g. dosed animals showed ischemic tubular necrosis in the kidney but only i.v. dosed animals developed adenocarcinomas of the mammary glands. Acutely dosed i.g. animals produced apoptotic bodies in the liver, necrosis of blood cell precursors in the bone marrow and ischemic tubular necrosis in the kidney but they did not develop tumors. No mutations were found in the H-ras and p53 genes in the mammary glands of either the i.g. rats or the tumor-bearing i.v. rats. However, the mammary glands of a fourth group of rats, which received APT by i.v. and killed before tumor development, carried Pu to Pu and Pu to Py double mutations in codons 58 and 59 of H-ras. This study indicates that the route of administration plays a role in the nature of the disease expression from ptaquiloside exposure. In addition to confirming the role of APT in the PT-induced carcinogenesis our finding suggests that activation of H-ras is an early event in the PT-carcinogenesis model. PMID- 9753660 TI - Presence of dioxins in human follicular fluid: their possible stage-specific action on the development of preimplantation mouse embryos. AB - Examination of human follicular fluid revealed the presence of polychlorinated dibenzodioxins (PCDDs) and dibenzofurans (PCDFs) at concentrations of approximately 1 pg/ml (0.01 pg TEQ/ml). To study their possible action, two-cell mouse embryos were cultured in the presence of 2,3,7,8-tetrachlorodibenzo-p dioxin (TCDD) at concentrations between 0.5 and 100 pM and evaluated at 24-h intervals for their development to the eight-cell and blastocyst stages. The percentage of eight-cell embryos exposed to TCDD at 1, 2, and 5 pM concentrations was significantly lower than that of controls. However, blastocyst formation of the surviving eight-cell embryos was accelerated, with the number of cells in the blastocysts increased in a dose-dependent manner. Findings suggest that PCDDs and PCDFs may be present in human reproductive fluid and may exert some stage specific effects on early embryonic development. PMID- 9753661 TI - The 77-kDa echinoderm microtubule-associated protein (EMAP) shares epitopes with the mammalian brain MAPs, MAP-2 and tau. AB - Previous work has shown that the echinoderm microtubule-associated protein (EMAP) was a unique MAP with little sequence similarity with the brain MAPs. The purpose of this study was to determine whether there were any small domains within EMAP that were shared by the mammalian brain MAPs, MAP-2, and tau. It is reported here that EMAP and the heat-stable MAP-2 and tau share antigenic determinants. A polyclonal antisera, raised against SDS-PAGE denatured EMAP, reacted strongly with both MAP-2 and tau on Western blots. In addition, a detailed sequence comparison, using a window of 5 amino acids at a time, revealed several short domains with approximately 20 residues that shared sequence similarity. The regions of sequence similarity were all located in regions implicated in microtubule binding, suggesting that EMAP and the mammalian brain MAPs may share short structural and functional domains. PMID- 9753662 TI - Carnitine biosynthesis: identification of the cDNA encoding human gamma butyrobetaine hydroxylase. AB - gamma-Butyrobetaine hydroxylase (EC 1.14.11.1) is the last enzyme in the biosynthetic pathway of L-carnitine and catalyzes the formation of L-carnitine from gamma-butyrobetaine, a reaction dependent on alpha-ketoglutarate, Fe2+, and oxygen. We report the purification of the protein from rat liver to apparent homogeneity, which allowed N-terminal sequencing using Edman degradation. The obtained amino acid sequence was used to screen the expressed sequence tag database and led to the identification of a human cDNA containing an open reading frame of 1161 base pairs encoding a polypeptide of 387 amino acids with a predicted molecular weight of 44.7 kDa. Heterologous expression of the open reading frame in the yeast Saccharomyces cerevisiae confirmed that the cDNA encodes the human gamma-butyrobetaine hydroxylase. Northern blot analysis showed gamma-butyrobetaine hydroxylase expression in kidney (high), liver (moderate), and brain (very low), while no expression could be detected in the other investigated tissues. PMID- 9753663 TI - Inhibition of L-type calcium channels by octreotide in isolated human neuroendocrine tumor cells of the gut. AB - The observation that somatostatin and its analogue octreotide inhibit the release of various peptide hormones and transmitters from neuroendocrine tumors has stimulated interest in the signal transduction pathway mediated by these compounds. Using the whole cell mode of the patch-clamp technique, we investigated the inhibitory effects of somatostatin and octreotide on voltage dependent calcium channels (VDCC) in isolated human neuroendocrine tumor cells of the gut. Both peptides dose dependently and reversibly inhibited VDCC. Somatostatin (100 nM) reduced the current amplitude by 38 +/- 19% and 100 nM octreotide by 35 +/- 14%. Human neuroendocrine gut tumor cells preferentially express dihydropyridine-sensitive L-type VDCC, since most of the inward current was sensitive to the dihydropyridine isradipine. The inhibitory effects of isradipine and octreotide were not additive and octreotide had little effect on the isradipine-resistant inward current. Since octrotide selectively binds to the somatostatin receptor subtypes 2 and 5, these results suggest that inhibition of calcium-dependent hormone release by somatostatin from human neuroendocrine gut cells appears to involve somatostatin receptor subtypes 2 and 5, as well as dihydropyridine-sensitive L-type VDCC. PMID- 9753664 TI - Transcriptional regulation of BMP-4 in the Xenopus embryo: analysis of genomic BMP-4 and its promoter. AB - Recent experiments in the Xenopus embryo suggest that proper regulation of BMP-4 signaling is critical to the dorsal ventral specification of both mesoderm and ectoderm. Regulation of BMP-4 signaling is known to occur extracellularly by direct binding with chordin, noggin, and follistatin, and intracellularly through the antagonistic signal interaction with dorsalizing TGF-beta family member activin. However, tight repressional regulation of BMP transcription may also be required to sustain the dorsal and neural status of the induced cells. Here we demonstrate that the dominant negative mutant of the BMP receptor (DN-BR) or the BMP-4 antagonizers, chordin and noggin, negatively regulate BMP-4 transcription in animal cap explants. We suggest that repression of BMP-4 transcription is important in the maintenance of dorsal fate and that continuous input of BMP-4 signaling is required to sustain the expression of BMP-4 transcription in the maintenance of epidermal/ventral fate. Consistent with this postulation, we found that the promoter region of the isolated BMP-4 genomic DNA includes several consensus binding sites for transcriptional regulators functioning under BMP-4 signaling such as GATA binding and ventralizing homeobox genes. In a functional assay we found that the GATA binding and ventral homeobox proteins can positively modulate BMP-4 promoter activity. We also observed that DN-BR decreases BMP-4 promoter activity. This was likely due to a repression of the above-mentioned transcription factors. The significance of these observations to embryonic patterning is discussed. PMID- 9753665 TI - Use of the glyceraldehyde-3-phosphate dehydrogenase promoter for production of functional mammalian membrane transport proteins in the yeast Pichia pastoris. AB - The promoter of the glyceraldehyde-3-phosphate dehydrogenase gene (PGAP) was employed to produce the mammalian peptide transporters hPEPT1 and rPEPT2 as models for polytopic transmembrane proteins in the methylotrophic yeast Prichia pastoris. Cells of a recombinant renal peptide transporter (rPEPT2) clone produced constitutively the functional carrier protein. The level of functional expression of rPEPT2 with PGAP varied depending on the carbon source used for cell growth, but was up to five times higher than that obtained with the commonly employed inducible alcohol oxidase 1 promoter (PAOX1). Similar results were obtained for the expression level of the human intestinal peptide transporter hPEPT1 controlled by either PGAP or PAOX1. Therefore, the PGAP seems to be an attractive alternative to PAOX1 for generation of transgenic P. pastoris cells expressing functional mammalian membrane transport proteins at high levels. PMID- 9753668 TI - The distribution of endogenous retinoic acid in the chick embryo: implications for developmental mechanisms. AB - The aim of these experiments was to determine the endogenous distribution of retinoic acid (RA) across a wide range of embryonic stages in the chick embryo. By high pressure liquid chromatography, it was revealed that didehydroRA is the most prevalent retinoic acid in the chick embryo and that the tissues of the stage 24 embryo differed widely in their total RA content (didehydroRA + all trans-RA). Some tissues such as the heart had very little RA and some such as the neural tube had very high levels, the total variation between these two being 29 fold. We showed that these tissues also synthesised RA and released it into the medium, thus validating the use of the F9 reporter cell system for further analyses of younger staged embryos. With these F9 cells, we showed that, at stage 4, the posterior end of the embryo had barely detectably higher levels of RA than the anterior end, but that a significant level of RA generation was detected as soon as somitogenesis began. Then a sharp on/off boundary of RA was present at the level of the first somite. We could find no evidence for a posterior-to anterior gradient of RA. Throughout further development, various consistent observations were made: the developing brain did not generate RA, but the spinal part of the neural tube generated it at very high levels so there must be a sharp on/off boundary in the region of the hindbrain/spinal cord junction; the mesenchyme surrounding the hindbrain generated RA whereas the hindbrain itself did not; there was a variation in RA levels from the midline outwards with the highest levels of RA in the spinal neural tube followed by lower levels in the somites followed by lower levels in the lateral plate; the posterior half of the limb bud generated higher levels than the anterior half. With these observations, we were able to draw maps of endogenous RA throughout these early stages of chick embryogenesis and the developmental implications of these results are discussed. PMID- 9753669 TI - The knirps and knirps-related genes organize development of the second wing vein in Drosophila. AB - The neighboring homologous knirps (kni) and knirps-related (knrl) genes in Drosophila encode transcription factors in the steroid hormone receptor superfamily. During early embryogenesis, kni functions as a gap gene to control expression of segmentation genes within the abdominal region of the embryo. In this study, we present evidence that kni and knrl link A/P positional information in larval wing imaginal discs to morphogenesis of the second longitudinal wing vein (L2). We show that kni and knrl are expressed in similar narrow stripes corresponding to the position of the L2 primordium. The kni and knrl L2 stripes abut the anterior border of the broad central expression domain of the Dpp target gene spalt major (salm). We provide evidence that radius incompletus (ri), a well known viable mutant lacking the L2 vein, is a regulatory mutant of the kni/knrl locus. In ri mutant wing discs, kni and knrl fail to be expressed in the L2 primordium. In addition, the positions of molecular breakpoints in the kni/knrl locus indicate that the ri function is provided by cis-acting sequences upstream of the kni transcription unit. Epistasis tests reveal that the kni/knrl locus functions downstream of spalt major (salm) and upstream of genes required to initiate vein-versus-intervein differentiation. Mis-expression experiments suggest that kni and knrl expressing cells inhibit neighboring cells from becoming vein cells. Finally, kni and knrl are likely to refine the L2 position by positively auto-regulating their own expression and by providing negative feedback to repress salm expression. We propose a model in which the combined activities of kni and knrl organize development of the L2 vein in the appropriate position. PMID- 9753670 TI - Differential activation of Myf5 and MyoD by different Wnts in explants of mouse paraxial mesoderm and the later activation of myogenesis in the absence of Myf5. AB - Activation of myogenesis in newly formed somites is dependent upon signals derived from neighboring tissues, namely axial structures (neural tube and notochord) and dorsal ectoderm. In explants of paraxial mesoderm from mouse embryos, axial structures preferentially activate myogenesis through a Myf5 dependent pathway and dorsal ectoderm preferentially through a MyoD-dependent pathway. Here we report that cells expressing Wnt1 will preferentially activate Myf5 while cells expressing Wnt7a will preferentially activate MyoD. Wnt1 is expressed in the dorsal neural tube and Wnt7a in dorsal ectoderm in the early embryo, therefore both can potentially act in vivo to activate Myf5 and MyoD, respectively. Wnt4, Wnt5a and Wnt6 exert an intermediate effect activating both Myf5 and MyoD equivalently in paraxial mesoderm. Sonic Hedgehog synergises with both Wnt1 and Wnt7a in explants from E8.5 paraxial mesoderm but not in explants from E9.5 embryos. Signaling through different myogenic pathways may explain the rescue of muscle formation in Myf5 null embryos, which do not form an early myotome but later develop both epaxial and hypaxial musculature. Explants of unsegmented paraxial mesoderm contain myogenic precursors capable of expressing MyoD in response to signaling from a neural tube isolated from E10.5 embryos, the developmental stage when MyoD is present throughout the embryo. Myogenic cells cannot activate MyoD in response to signaling from a less mature neural tube. Together these data suggest that different Wnt molecules can activate myogenesis through different pathways such that commitment of myogenic precursors is precisely regulated in space and time to achieve the correct pattern of skeletal muscle development. PMID- 9753671 TI - The Drosophila Pax gene eye gone is required for embryonic salivary duct development. AB - What are the developmental mechanisms required for conversion of an undifferentiated, two-dimensional field of cells into a patterned, tubular organ? In this report, we describe the contribution of the Drosophila Pax gene eye gone to the development of the embryonic salivary glands and ducts. eye gone expression in salivary tissues is controlled by several known regulators of salivary fate. After the initial establishment of the salivary primordium by Sex combs reduced, fork head excludes eye gone expression from the pregland cells so that its salivary expression is restricted to the posterior preduct cells. trachealess, in contrast, activates eye gone expression in the posterior preduct cells. We have previously described the process by which fork head and the EGF receptor pathway define the border between the gland and duct primordia. Here we show that eye gone is required for the subdivision of the duct primordium itself into the posterior individual duct and the anterior common duct domains. In the absence of eye gone, individual ducts as well as the precursor of the adult salivary glands, the imaginal ring, are absent. We took advantage of this ductless phenotype to show that Drosophila larvae do not have an obligate requirement for salivary glands and ducts. In addition to its role in the salivary duct, eye gone is required in the embryo for the development of the eye antennal imaginal disc and the chemosensory antennal organ. PMID- 9753672 TI - Involvement of the sonic hedgehog, patched 1 and bmp2 genes in patterning of the zebrafish dermal fin rays. AB - The signaling molecule encoded by Sonic hedgehog (shh) participates in the patterning of several embryonic structures including limbs. During early fin development in zebrafish, a subset of cells in the posterior margin of pectoral fin buds express shh. We have shown that regulation of shh in pectoral fin buds is consistent with a role in mediating the activity of a structure analogous to the zone of polarizing activity (ZPA) (Akimenko and Ekker (1995) Dev. Biol. 170, 243-247). During growth of the bony rays of both paired and unpaired fins, and during fin regeneration, there does not seem to be a region equivalent to the ZPA and one would predict that shh would play a different role, if any, during these processes specific to fish fins. We have examined the expression of shh in the developing fins of 4-week old larvae and in regenerating fins of adults. A subset of cells in the basal layer of the epidermis in close proximity to the newly formed dermal bone structures of the fin rays, the lepidotrichia, express shh, and ptc1 which is thought to encode the receptor of the SHH signal. The expression domain of ptc1 is broader than that of shh and adjacent blastemal cells releasing the dermal bone matrix also express ptc1. Further observations indicate that the bmp2 gene, in addition to being expressed in the same cells of the basal layer of the epidermis as shh, is also expressed in a subset of the ptc1-expressing cells of the blastema. Amputations of caudal fins immediately after the first branching point of the lepidotrichia, and global administration of all-trans-retinoic acid, two procedures known to cause fusion of adjacent rays, result in a transient decrease in the expression of shh, ptc1 and bmp2. The effects of retinoic acid on shh expression occur within minutes after the onset of treatment suggesting direct regulation of shh by retinoic acid. These observations suggest a role for shh, ptc1 and bmp2 in patterning of the dermoskeleton of developing and regenerating teleost fins. PMID- 9753673 TI - Targeting gene expression to the head: the Drosophila orthodenticle gene is a direct target of the Bicoid morphogen. AB - The Bicoid (Bcd) morphogen establishes the head and thorax of the Drosophila embryo. Bcd activates the transcription of identified target genes in the thoracic segments, but its mechanism of action in the head remains poorly understood. It has been proposed that Bcd directly activates the cephalic gap genes, which are the first zygotic genes to be expressed in the head primordium. It has also been suggested that the affinity of Bcd-binding sites in the promoters of Bcd target genes determines the posterior extent of their expression (the Gene X model). However, both these hypotheses remain untested. Here, we show that a small regulatory region upstream of the cephalic gap gene orthodenticle (otd) is sufficient to recapitulate early otd expression in the head primordium. This region contains two control elements, each capable of driving otd-like expression. The first element has consensus Bcd target sites that bind Bcd in vitro and are necessary for head-specific expression. As predicted by the Gene X model, this element has a relatively low affinity for Bcd. Surprisingly, the second regulatory element has no Bcd sites. Instead, it contains a repeated sequence motif similar to a regulatory element found in the promoters of otd related genes in vertebrates. Our study is the first demonstration that a cephalic gap gene is directly regulated by Bcd. However, it also shows that zygotic gene expression can be targeted to the head primordium without direct Bcd regulation. PMID- 9753674 TI - The expression and regulation of chick EphA7 suggests roles in limb patterning and innervation. AB - Eph receptors and their ligands, the ephrins, have been implicated in early patterning and axon guidance in vertebrate embryos. Members of these families play pivotal roles in the formation of topographic maps in the central nervous system, the formation of brain commissures, and in the guidance of neural crest cells and motor axons through the anterior half of the somites. Here, we report a highly dynamic expression pattern of the chick EphA7 gene in the developing limb. Expression is detected in discrete domains of the dorsal mesenchyme from 3 days of incubation. The expressing cells are adjacent to the routes where axons grow to innervate the limb at several key points: the region of plexus formation, the bifurcation between dorsal and ventral fascicles, and the pathway followed by axons innervating the dorsal muscle mass. These results suggested a role for EphA7 in cell-cell contact-mediated signalling in dorsal limb patterning and/or axon guidance. We carried out experimental manipulations in the chick embryo wing bud to alter the dorsoventral patterning of the limb. The analyses of EphA7 expression and innervation in the operated wings indicate that a signal emanating from the dorsal ectoderm regulates EphA7 in such a way that, in its absence, the wing bud lacks EphA7 expression and shows innervation defects at the regions where the gene was downregulated. EphA7 downregulation in the dorsal mesenchyme after dorsal ectoderm removal is more rapid than that of Lmx-1, the gene known to mediate dorsalisation in response to the ectodermal signal. These results add a new gene to the dorsalisation signalling pathway in the limb. Moreover, they implicate the Eph receptor family in the patterning and innervation of the developing limb, extending its role in axon pathfinding to the distal periphery. PMID- 9753675 TI - Avian neural crest-derived neurogenic precursors undergo apoptosis on the lateral migration pathway. AB - Neural crest cells of vertebrate embryos disperse on distinct pathways and produce different derivatives in specific embryonic locations. In the trunk of avian embryos, crest-derived cells that initially migrate on the lateral pathway, between epidermal ectoderm and somite, produce melanocytes but no neuronal derivatives. Although we found that melanocyte precursors are specified before they disperse on the lateral pathway, we also observed that a few crest-derived neuronal cells are briefly present on the same pathway. Here, we show that neuronal cells are removed by an episode of apoptosis. These observations suggest that localized environmental factor(s) affect the distribution of fate-restricted crest derivatives and function as a 'proof-reading mechanism' to remove 'ectopic' crest-derived cells. PMID- 9753676 TI - Role of dpp signalling in prepattern formation of the dorsocentral mechanosensory organ in Drosophila melanogaster. AB - A proneural cluster of dorsocentral bristles forms adjacent to the dorsal side of wg-expressing cells in the notum region of the wing imaginal disc. It has been shown that wg activity is required for these structures to form. However, the restriction of this proneural cluster to the dorsal posterior side of the wg expression domain in the anterior compartment of the wing imaginal disc has suggested that Wg signalling itself is insufficient to establish the dorsocentral proneural cluster. Some factor(s) from the posterior side must participate in this action in cooperation with Wg signalling. We have examined the role of Dpp signalling in dorsocentral bristle formation by either ectopically activating or conditionally reducing Dpp signalling. Ubiquitous activation of Dpp signalling in the notum region of the wing imaginal disc induced additional dorsocentral proneural cluster all along the dorsal side of the wg expression domain, and altered wg expression. Conditional loss-of-function of Dpp signalling during disc development resulted in the inhibition of dorsocentral proneural cluster formation and expansion of the wg expression domain. These results suggest that Dpp signalling has two indispensable roles in dorsocentral bristle formation: induction of the dorsocentral proneural cluster in cooperation with Wg signalling and restriction of the wg expression domain in the notum region of the wing imaginal disc. PMID- 9753677 TI - Wnt-4 is a mesenchymal signal for epithelial transformation of metanephric mesenchyme in the developing kidney. AB - Development of the mammalian kidney is initiated by ingrowth of the ureteric bud into the metanephric blastema. In response to signal(s) from the ureter, mesenchymal cells condense, aggregate into pretubular clusters, and undergo epithelialisation to form simple epithelial tubules. Subsequent morphogenesis and differentiation of the tubular epithelium lead to the establishment of a functional nephron. Here we demonstrate that Wnt-4, a secreted glycoprotein which is required for tubule formation, is sufficient to trigger tubulogenesis in isolated metanephric mesenchyme, whereas Wnt-11 which is expressed in the tip of the growing ureter is not. Wnt-4 signaling depends on cell contact and sulphated glycosaminoglycans and is only required for triggering tubulogenesis but not for later events. The Wnt-4 signal can be replaced by other members of the Wnt gene family including Wnt-1, Wnt-3a, Wnt-7a and Wnt-7b. Further, dorsal spinal cord, which has been thought to mimic ureteric signaling in tubule induction induces Wnt-4 mutant as well as wild-type mesenchyme suggesting that spinal cord derived signal(s) most likely act by mimicking the normal mesenchymal action of Wnt-4. These results lend additional support to the notion that Wnt-4 is a key auto regulator of the mesenchymal to epithelial transformation that underpins nephrogenesis adding another level of complexity in the hierarchy of molecular events mediating tubulogenesis. PMID- 9753678 TI - LAM1 is required for dorsoventrality and lateral growth of the leaf blade in Nicotiana. AB - The role of LAM1 in dorsoventrality and lateral growth of the leaf blade was investigated in the 'bladeless' lam1 mutant of Nicotiana sylvestris and in periclinal chimeras with lam1 and wild-type (N. glauca) cell layers. Mutant lam1 primordia show normal dorsoventrality at emergence, but produce blade tissue that lacks dorsal cell types and fails to expand in the lateral plane. In leaves of a lam1-glauca-glauca (L1-L2-L3) chimera, we observed restoration of dorsal identity in the lam1 upper epidermis, suggesting non-cell-autonomous movement of a dorsalizing factor between cell layers of the blade. A lam1-lam1-glauca chimera generated a leaf blade with lam1 cells in the L1-derived epidermis and the L2 derived upper and lower mesophyll. An in situ lineage analysis revealed that N. glauca cells in the L3-derived middle mesophyll restore palisade differentiation in the adjoining lam1 upper mesophyll. Movement of dorsalizing information appears short-range, however, having no effect on the upper lam1 epidermis in lam1-lam1-glauca. Clusters of lam1 mesophyll in distal or proximal positions show a localized default to radial growth, indicating that the LAM1 function is required for dorsoventrality and lateral growth throughout blade expansion. PMID- 9753679 TI - Boundaries in the Drosophila wing imaginal disc organize vein-specific genetic programs. AB - Previous studies have suggested that vein primordia in Drosophila form at boundaries along the A/P axis between discrete sectors of the larval wing imaginal disc. Genes involved in initiating vein development during the third larval instar are expressed either in narrow stripes corresponding to vein primordia or in broader 'provein' domains consisting of cells competent to become veins. In addition, genes specifying the alternative intervein cell fate are expressed in complementary intervein regions. The regulatory relationships between genes expressed in narrow vein primordia, in broad provein stripes and in interveins remains unknown, however. In this manuscript, we provide additional evidence for veins forming in narrow stripes at borders of A/P sectors. These experiments further suggest that narrow vein primordia produce secondary short range signal(s), which activate expression of provein genes in a broad pattern in neighboring cells. We also show that crossregulatory interactions among genes expressed in veins, proveins and interveins contribute to establishing the vein versus-intervein pattern, and that control of gene expression in vein and intervein regions must be considered on a stripe-by-stripe basis. Finally, we present evidence for a second set of vein-inducing boundaries lying between veins, which we refer to as paravein boundaries. We propose that veins develop at both vein and paravein boundaries in more 'primitive' insects, which have up to twice the number of veins present in Drosophila. We present a model in which different A/P boundaries organize vein-specific genetic programs to govern the development of individual veins. PMID- 9753680 TI - The cellular mechanism by which the dermomyotome contributes to the second wave of myotome development. AB - We have shown that a subset of early postmitotic progenitors that originates along the medial part of the epithelial somite gives rise to the primary myotome (Kahane, N., Cinnamon, Y. and Kalcheim, C. (1998). Mech. Dev. 74, 59-73). Because of its postmitotic nature, further myotome expansion must be achieved by cell addition from extrinsic sources. Here we investigate the mechanism whereby the dermomyotome contributes to this process. Using several different methods we found that cell addition occurs from both rostral and caudal edges of the dermomyotome, but not directly from its dorsomedial lip (DML). First, labeling of quail embryos with [3H]thymidine revealed a time-dependent entry of radiolabeled nuclei into the myotome from the entire rostral and caudal lips of the dermomyotome, but not from the DML. Second, fluorescent vital dyes were injected at specific sites in the dermomyotome lips and the fate of dye-labeled cells followed by confocal microscopy. Consistent with the nucleotide labeling experiments, dye-labeled myofibers directly emerged from injected epithelial cells from either rostral or caudal lips. In contrast, injected cells from the DML first translocated along the medial boundary, reached the rostral or caudal dermomyotome lips and only then elongated into the myotome. These growing myofibers had always one end attached to either lip from which they elongated in the opposite direction. Third, following establishment of the primary myotome, cells along the extreme dermomyotome edges, but not the DML, expressed QmyoD, supporting the notion that rostral and caudal boundaries generate myofibers. Fourth, ablation of the DML had only a limited effect on myotomal cell number. Thus, cells deriving from the extreme dermomyotome lips contribute to uniform myotome growth in the dorsoventral extent of the myotome. They also account for its expansion in the transverse plane and this is achieved by myoblast addition in a lateral to medial direction (from the dermal to the sclerotomal sides), restricting the pioneer myofibers to the dermal side of the myotome. Taken together, the data suggest that myotome formation is a multistage process. A first wave of pioneers establishes the primary structure. A second wave generated from specific dermomyotome lips contributes to its expansion. Because dermomyotome lip progenitors are mitotically active within the epithelia of origin but exit the cell cycle upon myotome colonization, they can only provide for limited myotome growth and subsequent waves must take over to ensure further muscle development. PMID- 9753681 TI - A potential role for the OTX2 homeoprotein in creating early 'highways' for axon extension in the rostral brain. AB - Brain pattern formation starts with a subdivision of the neuroepithelium through site-specific expression of regulatory genes and, subsequently, the boundaries between presumptive neuromeres may provide a scaffold for early formation of axon tracts. In the mouse forebrain, the transcription factor OTX2 is strongly expressed at several such boundaries. Combining dye tracing and staining for OTX2 protein, we show that a number of early fibre tracts develop within stripes of OTX2 expression. To analyse a putative influence of OTX2 on the expression of molecules involved in neurite growth, we generated several clones of NIH3T3 cells stably expressing OTX2 protein at varying levels. As shown by immunoblotting, Otx2 transfection affects the expression of a variety of cell and substratum adhesion molecules, rendering the cells a favourable substratum in neurite outgrowth assays. Among the molecules upregulated with increasing levels of OTX2 are NCAM, tenascin-C and DSD-1-PG, which also in situ colocalize with zones of OTX2 expression at boundaries. These data suggest that Otx2 might be involved in defining local substrata for axon extension in the forebrain. PMID- 9753682 TI - Evidence for a frizzled-mediated wnt pathway required for zebrafish dorsal mesoderm formation. AB - We have used zebrafish as a model system for the study of vertebrate dorsoventral patterning. We isolated a maternally expressed and dorsal organizer localized member of the frizzled family of wnt receptors. Wild-type and dominant, loss-of function molecules in misexpression studies demonstrate frizzled function is necessary and sufficient for dorsal mesoderm specification. frizzled activity is antagonized by the action of GSK-3, and we show GSK-3 is also required for zebrafish dorsal mesoderm formation. frizzled cooperatively interacts with the maternally encoded zebrafish wnt8 protein in dorsal mesodermal fate determination. This frizzled -mediated wnt pathway for dorsal mesoderm specification provides the first evidence for the requirement of a wnt-like signal in vertebrate axis determination. PMID- 9753683 TI - Expression of calcitonin receptors in mouse preimplantation embryos and their function in the regulation of blastocyst differentiation by calcitonin. AB - Calcitonin secretion in the pregnant uterus is tightly regulated by the ovarian hormones, estrogen and progesterone, which limit its expression to a brief period preceding blastocyst implantation. The binding of calcitonin to a G protein coupled receptor activates adenylate cyclase and elevates cytosolic Ca2+ levels. The acceleration of preimplantation embryonic development that is known to occur upon elevation of intracellular Ca2+ prompted an investigation into calcitonin regulation of blastocyst differentiation. Using reverse transcription and the polymerase chain reaction to estimate the relative abundance of calcitonin receptor mRNA, a 25-fold accumulation of the splice variant, CR-1a, was observed in embryos between the 1-cell and 8-cell stages. Cytosolic free Ca2+ levels were rapidly elevated in embryos at the 4-cell to blastocyst stages after exposure to 10 nM calcitonin. Blastocysts treated for 30 minutes with 10 nM calcitonin differentiated in vitro at an accelerated rate, as assessed by the translocation of alpha5beta1 integrin to the apical surface of trophoblast cells, the corresponding elevation of fibronectin-binding activity and the timing of trophoblast cell migration. Chelation of cytosolic free Ca2+ with BAPTA-AM, but not inhibition of protein kinase A activity by H-89, attenuated the effects of calcitonin on blastocyst development. These findings support the concept that calcitonin secretion within the progesterone-primed uterus and the coordinate expression of CR-1a by preimplantation embryos regulates blastocyst differentiation through receptor-mediated Ca2+ signaling. PMID- 9753684 TI - Non-autonomy of AGAMOUS function in flower development: use of a Cre/loxP method for mosaic analysis in Arabidopsis. AB - Angiosperms use a multi-layered meristem (typically L1, L2 and L3) to produce primordia that then develop into plant organs. A number of experiments show that communication between the cell layers is important for normal development. We examined whether the function of the flower developmental control gene AGAMOUS involves communication across these layers. We developed a mosaic strategy using the Cre/loxP site-specific recombinase system, and identified the sector structure for mosaics that produced mutant flowers. The major conclusions were that (1) AGAMOUS must be active in the L2 for staminoid and carpelloid tissues, (2) that AGAMOUS must be active in the L2 and the L3 for floral meristem determinacy, and (3) that epidermal cell identity can be communicated by the L2 to the L1 layer. PMID- 9753685 TI - Semaphorins III and IV repel hippocampal axons via two distinct receptors. AB - The semaphorins are the largest family of repulsive axon guidance molecules. Secreted semaphorins bind neuropilin receptors and repel sensory, sympathetic and motor axons. Here we show that CA1, CA3 and dentate gyrus axons from E15-E17 mouse embryo explants are selectively repelled by entorhinal cortex and neocortex. The secreted semaphorins Sema III and Sema IV and their receptors Neuropilin-1 and -2 are expressed in the hippocampal formation during appropriate stages. Sema III and Sema IV strongly repel CA1, CA3 and dentate gyrus axons; entorhinal axons are only repelled by Sema III. An antibody against Neuropilin-1 blocks the repulsive action of Sema III and the entorhinal cortex, but has no effect on Sema IV-induced repulsion. Thus, chemorepulsion plays a role in axon guidance in the hippocampus, secreted semaphorins are likely to be responsible for this action, and the same axons can be repelled by two distinct semaphorins via two different receptors. PMID- 9753686 TI - FGFs and BMP4 induce both Msx1-independent and Msx1-dependent signaling pathways in early tooth development. AB - During early tooth development, multiple signaling molecules are expressed in the dental lamina epithelium and induce the dental mesenchyme. One signal, BMP4, has been shown to induce morphologic changes in dental mesenchyme and mesenchymal gene expression via Msx1, but BMP4 cannot substitute for all the inductive functions of the dental epithelium. To investigate the role of FGFs during early tooth development, we examined the expression of epithelial and mesenchymal Fgfs in wild-type and Msx1 mutant tooth germs and tested the ability of FGFs to induce Fgf3 and Bmp4 expression in wild-type and Msx1 mutant dental mesenchymal explants. Fgf8 expression is preserved in Msx1 mutant epithelium while that of Fgf3 is not detected in Msx1 mutant dental mesenchyme. Moreover, dental epithelium as well as beads soaked in FGF1, FGF2 or FGF8 induce Fgf3 expression in dental mesenchyme in an Msx1-dependent manner. These results indicate that, like BMP4, FGF8 constitutes an epithelial inductive signal capable of inducing the expression of downstream signaling molecules in dental mesenchyme via Msx1. However, the BMP4 and FGF8 signaling pathways are distinct. BMP4 cannot induce Fgf3 nor can FGFs induce Bmp4 expression in dental mesenchyme, even though both signaling molecules can induce Msx1 and Msx1 is necessary for Fgf3 and Bmp4 expression in dental mesenchyme. In addition, we have investigated the effects of FGFs and BMP4 on the distal-less homeobox genes Dlx1 and Dlx2 and we have clarified the relationship between Msx and Dlx gene function in the developing tooth. Dlx1,Dlx2 double mutants exhibit a lamina stage arrest in maxillary molar tooth development (Thomas B. L., Tucker A. S., Qiu M. , Ferguson C. A., Hardcastle Z., Rubenstein J. L. R. and Sharpe P. T. (1997) Development 124, 4811 4818). Although the maintenance of molar mesenchymal Dlx2 expression at the bud stage is Msx1-dependent, both the maintenance of Dlx1 expression and the initial activation of mesenchymal Dlx1 and Dlx2 expression during the lamina stage are not. Moreover, in contrast to the tooth bud stage arrest observed in Msx1 mutants, Msx1,Msx2 double mutants exhibit an earlier phenotype closely resembling the lamina stage arrest observed in Dlx1,Dlx2 double mutants. These results are consistent with functional redundancy between Msx1 and Msx2 in dental mesenchyme and support a model whereby Msx and Dlx genes function in parallel within the dental mesenchyme during tooth initiation. Indeed, as predicted by such a model, BMP4 and FGF8, epithelial signals that induce differential Msx1 and Msx2 expression in dental mesenchyme, also differentially induce Dlx1 and Dlx2 expression, and do so in an Msx1-independent manner. These results integrate Dlx1, Dlx2 and Fgf3 and Fgf8 into the odontogenic regulatory hierarchy along with Msx1, Msx2 and Bmp4, and provide a basis for interpreting tooth induction in terms of transcription factors which, individually, are necessary but not sufficient for the expression of downstream signals and therefore must act in specific combinations. PMID- 9753688 TI - Looking to the horizon. PMID- 9753687 TI - The sacral neural crest contributes neurons and glia to the post-umbilical gut: spatiotemporal analysis of the development of the enteric nervous system. AB - The majority of the enteric nervous system is derived from vagal neural crest cells (NCC), which migrate to the developing gut, proliferate, form plexuses and differentiate into neurons and glia. However, for some time, controversy has existed as to whether cells from the sacral region of the neural crest also contribute to the enteric nervous system. The aim of this study was to investigate the spatiotemporal migration of vagal and sacral NCC within the developing gut and to determine whether the sacral neural crest contributes neurons and glia to the ENS. We utilised quail-chick chimeric grafting in conjunction with antibody labelling to identify graft-derived cells, neurons and glia. We found that vagal NCC migrated ventrally within the embryo and accumulated in the caudal branchial arches before entering the pharyngeal region and colonising the entire length of the gut in a proximodistal direction. During migration, vagal crest cells followed different pathways depending on the region of the gut being colonised. In the pre-umbilical intestine, NCC were evenly distributed throughout the splanchnopleural mesenchyme while, in the post umbilical intestine, they occurred adjacent to the serosal epithelium. Behind this migration front, NCC became organised into the presumptive Auerbach's and Meissner's plexuses situated on either side of the developing circular muscle layer. The colorectum was found to be colonised in a complex manner. Vagal NCC initially migrated within the submucosa, internal to the circular muscle layer, before migrating outwards, adjacent to blood vessels, towards the myenteric plexus region. In contrast, sacral NCC, which also formed the entire nerve of Remak, were primarily located in the presumptive myenteric plexus region and subsequently migrated inwards towards the submucosal ganglia. Although present throughout the post-umbilical gut, sacral NCC were most numerous in the distal colorectum where they constituted up to 17% of enteric neurons, as identified by double antibody labelling using the quail-cell-specific marker, QCPN and the neuron-specific marker, ANNA-1. Sacral NCC were also immunopositive for the glial specific antibody, GFAP, thus demonstrating that this region of the neural crest contributes neurons and glia to the enteric nervous system. PMID- 9753689 TI - An excitatory scorpion toxin with a distinctive feature: an additional alpha helix at the C terminus and its implications for interaction with insect sodium channels. AB - BACKGROUND: Scorpion neurotoxins, which bind and modulate sodium channels, have been divided into two groups, the alpha and beta toxins, according to their activities. The beta-toxin class includes the groups of excitatory and depressant toxins, which differ in their mode of action and are highly specific against insects. The three-dimensional structures of several alpha and beta toxins have been determined at high resolution, but no detailed 3D structure of an excitatory toxin has been presented so far. RESULTS: The crystal structure of an anti-insect excitatory toxin from the scorpion Buthotus judaicus, Bj-xtrIT, has been determined at 2.1 A resolution and refined to an R factor of 0.209. The first 59 residues form a closely packed module, structurally similar to the conserved alpha and beta toxins ('long toxins') affecting sodium channels. The last 17 residues form a C-terminal extension not previously seen in scorpion toxins. It comprises a short alpha helix anchored to the N-terminal module by a disulfide bridge and is followed by a highly mobile stretch of seven residues, of which only four are seen in the electron-density map. This mobile peptide covers part of a conserved hydrophobic surface that is thought to be essential for interaction with the channel in several long toxins. CONCLUSIONS: Replacement of the last seven residues by a single glycine abolishes the activity of Bj-xtrIT, strongly suggesting that these residues are intimately involved in the interaction with the channel. Taken together with the partial shielding of the conserved hydrophobic surface and the proximity of the C terminus to an adjacent surface rich in charged residues, it seems likely that the bioactive surface of Bj-xtrIT is formed by residues surrounding the C terminus. The 3D structure and a recently developed expression system for Bj-xtrIT pave the way for identifying the structural determinants involved in the bioactivity and anti-insect specificity of excitatory toxins. PMID- 9753690 TI - The crystal structure of human cytosolic serine hydroxymethyltransferase: a target for cancer chemotherapy. AB - BACKGROUND: Serine hydroxymethyltransferase (SHMT) is a ubiquitous enzyme found in all prokaryotes and eukaryotes. As an enzyme of the thymidylate synthase metabolic cycle, SHMT catalyses the retro-aldol cleavage of serine to glycine, with the resulting hydroxymethyl group being transferred to tetrahydrofolate to form 5, 10-methylene-tetrahydrofolate. The latter is the major source of one carbon units in metabolism. Elevated SHMT activity has been shown to be coupled to the increased demand for DNA synthesis in rapidly proliferating cells, particularly tumour cells. Consequently, the central role of SHMT in nucleotide biosynthesis makes it an attractive target for cancer chemotherapy. RESULTS: We have solved the crystal structure of human cytosolic SHMT by multiple isomorphous replacement to 2.65 A resolution. The monomer has a fold typical for alpha class pyridoxal 5'-phosphate (PLP) dependent enzymes. The tetramer association is best described as a 'dimer of dimers' where residues from both subunits of one 'tight' dimer contribute to the active site. CONCLUSIONS: The crystal structure shows the evolutionary relationship between SHMT and other alpha class PLP-dependent enzymes, as the fold is highly conserved. Many of the results of site-directed mutagenesis studies can easily be rationalised or re-interpreted in light of the structure presented here. For example, His 151 is not the catalytic base, contrary to the findings of others. A mechanism for the cleavage of serine to glycine and formaldehyde is proposed. PMID- 9753691 TI - Structural basis of inhibitor selectivity in MAP kinases. AB - BACKGROUND: The mitogen-activated protein (MAP) kinases are important signaling molecules that participate in diverse cellular events and are potential targets for intervention in inflammation, cancer, and other diseases. The MAP kinase p38 is responsive to environmental stresses and is involved in the production of cytokines during inflammation. In contrast, the activation of the MAP kinase ERK2 (extracellular-signal-regulated kinase 2) leads to cellular differentiation or proliferation. The anti-inflammatory agent pyridinylimidazole and its analogs (SB [SmithKline Beecham] compounds) are highly potent and selective inhibitors of p38, but not of the closely-related ERK2, or other serine/threonine kinases. Although these compounds are known to bind to the ATP-binding site, the origin of the inhibitory specificity toward p38 is not clear. RESULTS: We report the structural basis for the exceptional selectivity of these SB compounds for p38 over ERK2, as determined by comparative crystallography. In addition, structural data on the origin of olomoucine (a better inhibitor of ERK2) selectivity are presented. The crystal structures of four SB compounds in complex with p38 and of one SB compound and olomoucine in complex with ERK2 are presented here. The SB inhibitors bind in an extended pocket in the active site and are complementary to the open domain structure of the low-activity form of p38. The relatively closed domain structure of ERK2 is able to accommodate the smaller olomoucine. CONCLUSIONS: The unique kinase-inhibitor interactions observed in these complexes originate from amino-acid replacements in the active site and replacements distant from the active site that affect the size of the domain interface. This structural information should facilitate the design of better MAP-kinase inhibitors for the treatment of inflammation and other diseases. PMID- 9753692 TI - A novel deamido-NAD+-binding site revealed by the trapped NAD-adenylate intermediate in the NAD+ synthetase structure. AB - BACKGROUND: Nicotinamide adenine dinucleotide (NAD+) has a central role in life processes. The ubiquitous enzyme NAD+ synthetase catalyzes a key step in NAD+ biosynthesis, transforming deamido-NAD+ into NAD+ by a two-step reaction. NAD+ synthetase belongs to the amidotransferase family and has been recognized as a member of the family of N-type ATP pyrophosphatases. In order to investigate the mechanism of the reaction carried out by NAD+ synthetase we have determined a high-resolution three-dimensional structure of the Bacillus subtilis homodimeric NAD+ synthetase in complex with the trapped reaction intermediate NAD-adenylate. RESULTS: Two NAD-adenylate molecules and two pyrophosphate (PPi) molecules are observed in the 1.3 A resolution structure of the NAD+ synthetase-NAD-adenylate complex. Structural studies on the NAD+ synthetase-NAD-adenylate adduct and on the cation-binding sites reveal a new deamido-NAD+-binding site located at the subunit interface, locate a binuclear magnesium cluster at the ATP-binding site and, identify two monovalent cation sites, one of which may represent an ammonium binding site. CONCLUSIONS: Our results suggest that two different catalytic strategies have been adopted by NAD+ synthetase in the two different steps of the reaction. During the adenylation step, no protein residues seem to be located properly to directly participate in catalysis, which is likely to be carried out with the fundamental assistance of an electron-withdrawing trimetallic constellation present in the active site. A different behavior is observed for the second step, in which an ammonium ion is the binding species. In this step, Asp173 is a key residue in both deprotonation of the primarily bound ammonium ion, and stabilization of the tetrahedral transition-state intermediate. Moreover, the structural data suggest that product release can take place only after all substrates are bound to the enzyme, and product release is ultimately controlled by the conformation adopted by two mobile loops. PMID- 9753693 TI - Mass spectrometric and thermodynamic studies reveal the role of water molecules in complexes formed between SH2 domains and tyrosyl phosphopeptides. AB - BACKGROUND: SH2 domains have a fundamental role in signal transduction. These domains interact with proteins containing phosphorylated tyrosine residues and, in doing so, mediate the interactions of proteins involved in tyrosine kinase signalling. The issue of specificity in SH2 domain interactions is therefore of great interest in terms of understanding tyrosine kinase signal-transduction pathways and in the discovery of drugs to inhibit them. Water molecules are found at the interfaces of many complexes, however, to date little attention has been paid to their role in dictating specificity. RESULTS: Here we use a combination of nanoflow electrospray ionization mass spectrometry (ESI-MS), isothermal titration calorimetry and structural data to investigate the effect of water molecules in complexes formed between the SH2 domain of tyrosine kinase Src and tyrosyl phosphopeptides. Binding studies have been performed using a series of different peptides that were selected to allow changes in the water content at the complex interface and demonstrate changes in specificity. ESI-MS enables quantification of the number of water molecules that interact with a higher affinity than those generally found solvating the biomolecular complex. CONCLUSIONS: Comparing the interactions of different peptides, we show that an intricate network of water molecules have a key role in dictating specificity. The use of mass spectrometry to quantify tightly bound water molecules may prove of general use in structural biology, where an independent determination of the water molecules associated with a structure would be advantageous. Furthermore, the ability to assess whether given water molecules are important in high affinity binding could make this method a precious tool in drug design. PMID- 9753694 TI - VEGF and the Fab fragment of a humanized neutralizing antibody: crystal structure of the complex at 2.4 A resolution and mutational analysis of the interface. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) is a highly specific angiogenic growth factor; anti-angiogenic treatment through inhibition of receptor activation by VEGF might have important therapeutic applications in diseases such as diabetic retinopathy and cancer. A neutralizing anti-VEGF antibody shown to suppress tumor growth in an in vivo murine model has been used as the basis for production of a humanized version. RESULTS: We present the crystal structure of the complex between VEGF and the Fab fragment of this humanized antibody, as well as a comprehensive alanine-scanning analysis of the contact residues on both sides of the interface. Although the VEGF residues critical for antibody binding are distinct from those important for high-affinity receptor binding, they occupy a common region on VEGF, demonstrating that the neutralizing effect of antibody binding results from steric blocking of VEGF receptor interactions. Of the residues buried in the VEGF-Fab interface, only a small number are critical for high-affinity binding; the essential VEGF residues interact with those of the Fab fragment, generating a remarkable functional complementarity at the interface. CONCLUSIONS: Our findings suggest that the character of antigen-antibody interfaces is similar to that of other protein protein interfaces, such as ligand-receptor interactions; in the case of VEGF, the principal difference is that the residues essential for binding to the Fab fragment are concentrated in one continuous segment of polypeptide chain, whereas those essential for binding to the receptor are distributed over four different segments and span across the dimer interface. PMID- 9753695 TI - Structural basis of activity and subunit recognition in G protein heterotrimers. AB - BACKGROUND: Inactive heterotrimeric G proteins are composed of a GDP-bound alpha subunit (Galpha) and a stable heterodimer of Gbeta and Ggamma subunits. Upon stimulation by a receptor, Galpha subunits exchange GDP for GTP and dissociate from Gbetagamma, both Galpha and Gbetagamma then interact with downstream effectors. Isoforms of Galpha, Gbeta and Ggamma potentially give rise to many heterotrimeric combinations, limited in part by amino acid sequence differences that lead to selective interactions. The mechanism by which GTP promotes Gbetagamma dissociation is incompletely understood. The Gly203-->Ala mutant of Gialpha1 binds and hydrolyzes GTP normally but does not dissociate from Gbetagamma, demonstrating that GTP binding and activation can be uncoupled. Structural data are therefore important for understanding activation and subunit recognition in G protein heterotrimers. RESULTS: The structures of the native (Gialpha1beta1gamma2) heterotrimer and that formed with Gly203-->AlaGialpha1 have been determined to resolutions of 2.3 A and 2.4 A, respectively, and reveal previously unobserved segments at the Ggamma2 C terminus. The Gly203-->Ala mutation alters the conformation of the N terminus of the switch II region (Val201-Ala203), but not the global structure of the heterotrimer. The N termini of Gbeta and Ggamma form a rigid coiled coil that packs at varying angles against the beta propeller of Gbeta. Conformational differences in the CD loop of beta blade 2 of Gbeta mediate isoform-specific contacts with Galpha. CONCLUSIONS: The Gly203-->Ala mutation in Gialpha1 blocks the conformational changes in switch II that are required to release Gbetagamma upon binding GTP. The interface between the ras-like domain of Galpha and the beta propeller of Gbeta appears to be conserved in all G protein heterotrimers. Sequence variation at the Gbeta-Galpha interface between the N-terminal helix of Galpha and the CD loop of beta blade 2 of Gbeta1 (residues 127-135) could mediate isoform-specific contacts. The specificity of Gbeta and Ggamma interactions is largely determined by sequence variation in the contact region between helix 2 of Ggamma and the surface of Gbeta. PMID- 9753696 TI - Amino-acid sequence and three-dimensional structure of the Staphylococcus aureus metalloproteinase at 1.72 A resolution. AB - BACKGROUND: Aureolysin is an extracellular zinc-dependent metalloproteinase from the pathogenic bacterium Staphylococcus aureus. This enzyme exhibits in vitro activity against several molecules of biological significance for the host, indicating that it is involved in the pathology of staphylococcal diseases. RESULTS: Here we report the amino-acid sequence and inhibitor-free X-ray crystal structure of aureolysin, a member of the thermolysin family of zinc-dependent metalloproteinases. This enzyme, which binds one zinc and three calcium ions, comprises a single chain of 301 amino acids that consists of a beta-strand-rich upper domain and an alpha-helix-rich lower domain. CONCLUSIONS: The overall structure of aureolysin is very similar to that of the other three members of this family whose structures are known - thermolysin (TLN) from Bacillus thermoproteolyticus, neutral protease (NP) from Bacillus cereus and elastase (PAE) from Pseudomonas aeruginosa. But an important difference has been encountered: in contrast to what has been observed in the other three members of this family (TLN, NP and PAE), inhibitor-free aureolysin displays a 'closed' active site cleft conformation. This new structure therefore raises questions about the universality of the hinge-bending motion model for the neutral metalloproteinases. PMID- 9753697 TI - CAMPASS: a database of structurally aligned protein superfamilies. PMID- 9753698 TI - The crystal structure of the novel snake venom plasminogen activator TSV-PA: a prototype structure for snake venom serine proteinases. AB - BACKGROUND: Trimeresurus stejnejeri venom plasminogen activator (TSV-PA) is a snake venom serine proteinase that specifically activates plasminogen. Snake venom serine proteinases form a subfamily of trypsin-like proteinases that are characterised by a high substrate specificity and resistance to inhibition. Many of these venom enzymes specifically interfere with haemostatic mechanisms and display a long circulating half-life. For these reasons several of them have commercial applications and are potentially attractive pharmacological tools. RESULTS: The crystal structure of TSV-PA has been determined to 2.5 A resolution and refined to an R factor of 17.8 (R free, 24.4). The enzyme, showing the overall polypeptide fold of trypsin-like serine proteinases, displays unique structural elements such as the presence of a phenylalanine at position 193, a C terminal tail clamped via a disulphide bridge to the 99-loop, and a structurally conserved Asp97 residue. The presence of a cis proline at position 218 is in agreement with evolutionary relationships to glandular kallikrein. CONCLUSIONS: We postulate that Phe 193 accounts for the high substrate specificity of TSV-PA and renders it incapable of forming a stable complex with bovine pancreatic trypsin inhibitor and other extended substrates and inhibitors. Mutational studies previously showed that Asp97 is crucial for the plasminogenolytic activity of TSV-PA, here we identify the conservation of Asp97 in both types of mammalian plasminogen activator - tissue-type (tPA) and urokinase-type (uPA). It seems likely that Asp97 of tPA and uPA will have a similar role in plasminogen recognition. The C-terminal extension of TSV-PA is conserved among snake venom serine proteinases, although its function is unknown. The three-dimensional structure presented here is the first of a snake venom serine proteinase and provides an excellent template for modelling other homologous family members. PMID- 9753699 TI - Structure of translation initiation factor 5A from Pyrobaculum aerophilum at 1.75 A resolution. AB - BACKGROUND: Translation initiation factor 5A (IF-5A) is reported to be involved in the first step of peptide bond formation in translation, to be involved in cell-cycle regulation and to be a cofactor for the Rev and Rex transactivator proteins of human immunodeficiency virus-1 and T-cell leukemia virus I, respectively. IF-5A contains an unusual amino acid, hypusine (N-epsilon-(4 aminobutyl-2-hydroxy)lysine), that is required for its function. The first step in the post-translational modification of lysine to hypusine is catalyzed by the enzyme deoxyhypusine synthase, the structure of which has been published recently. RESULTS: IF-5A from the archebacterium Pyrobaculum aerophilum has been heterologously expressed in Escherichia coli with selenomethionine substitution. The crystal structure of IF-5A has been determined by multiwavelength anomalous diffraction and refined to 1.75 A. Unmodified P. aerophilum IF-5A is found to be a beta structure with two domains and three separate hydrophobic cores. CONCLUSIONS: The lysine (Lys42) that is post-translationally modified by deoxyhypusine synthase is found at one end of the IF-5A molecule in an turn between beta strands beta4 and beta5; this lysine residue is freely solvent accessible. The C-terminal domain is found to be homologous to the cold-shock protein CspA of E. coli, which has a well characterized RNA-binding fold, suggesting that IF-5A is involved in RNA binding. PMID- 9753700 TI - A close family resemblance: the importance of structure in understanding cytochromes P450. AB - Cytochromes P450 comprise a very large superfamily of hemeproteins which generally monooxygenate hydrophobic compounds. P450s appear to have a common conserved structural core, yet are variable in regions involved in substrate recognition and binding, and in redox-partner binding. These differences can be identified by an analysis in which structural alignments and homology models are used to compare the various classes and families of P450s. PMID- 9753701 TI - Validation of an EEG-derived spectral frequency index (SFx) for continuous monitoring of sleep depth in humans. AB - Sleep in humans is classically assessed by recording a multichannel electroencephalogram (EEG) in connection with an electrooculogram (EOG) and an electromyogram (EMG). In general, human sleep is manually staged into 6 categories (from awake through REM sleep to stage 4 reflecting deep sleep) on the basis of a visual inspection of EEG periods (epochs) of 20 - 30 s duration. This cumbersome methodology is still used in practice and for reference purposes. - The conversion of EEG-signals by means of Fast Fourier Transformation provides objective and reproducible information reflecting specific communicative features of the central nervous system. A special part of this information based on a specific algorithm is defined by the so-called spectral frequency index (SFx). This SFx-algorithm contains relationships among some particular EEG frequencies and provides objective percentage values about the state of consciousness of a person. In order to validate this new SFx-method, sleep as a physiological state of continuous alterations of consciousness and vigilance was chosen. A total of 36 nights of sleep from 18 healthy volunteers were staged manually by a scientist unaware of the protocol. The volunteers received either placebo or lormetazepam prior to commencement of the nocturnal recordings. The manually staged data were compared with the data obtained by the SFx-analysis. Both data sets SFx values and manually staged data were made comparable by averaging their values to a basic period length of 2 min duration giving 7960 pairs of data. The SFx data for sleep were found within a range from 35% to 100%. The SFx-medians of the manually staged data from "awake" to stage 4 were found in a decending order ("awake": 83% (lower and upper quartile 78% / 87%);"REM": 68% (63%/74%),"stage 1" :63% (57%/70%),"stage 2" :51% (47%/57%), "stage 3" :44% (42%/46%) and "stage 4" :42% (40%/44%). The rank correlation coefficient between the data pairs was calculated to be 0.79 indicating a substantial matching between the manually staged score and the SFx. We therefore conclude that the SFx is a suitable and objective indicator of sleep depth in humans. PMID- 9753702 TI - Suppression of HHV-8 viremia by foscarnet in an HIV-infected patient with Kaposi's sarcoma and HHV-8 associated hemophagocytic syndrome. AB - Human herpes virus 8 (HHV-8) seems to be involved in the pathogenesis of Kaposi s sarcoma. In vitro, antiviral drugs with activity against herpes viruses also can suppress HHV-8, however, little is known about the antiviral activity against HHV 8 in vivo. In this report we describe the effects of foscarnet on HHV-8 viremia in an HIV-infected patient with disseminated Kaposi s sarcoma and a presumably HHV-8 associated hemophagocytic syndrome. HHV-8 DNA could be detected in this patient by PCR in peripheral blood mononuclear cells (PBMC), in bronchoalveolar fluid and tumor biopsies. After initiation of foscarnet because of a severe hemophagocytic syndrome HHV-8 PCR turned negative in PBMC, but stayed positive in pleural effusions and in a tumor biopsy. After termination of foscarnet therapy HHV-8 DNA in PBMC persistently reappeared. Under treatment with foscarnet the hemophagocytic syndrome dramatically improved, suggesting that HHV-8 had a pathogenetic role in this syndrome. PMID- 9753703 TI - Changes in platelet membrane glycoproteins before bone marrow transplantation and after engraftment--a pilot study. AB - Thrombotic complications are observed in patients undergoing bone marrow transplantation despite thrombocytopenia and impaired coagulation due to liver function disturbances. Endothelial cell damage which is involved in the pathogenesis of major transplant related complications like graft-versus-host disease, veno-occlusive disease, sepsis or microangiopathy may be a contributing factor. Little is known about platelet function in bone marrow transplant recipients. In order to study functional alterations in circulating platelets we investigated unstimulated and ADP-stimulated platelets of 10 bone marrow transplant recipients ex vivo by flow cytometry in a pilot study using a panel of monoclonal antibodies to characterize changes in membrane glycoproteins. Samples were collected before and during conditioning and at three timepoints after engraftment. 10 healthy volunteers served as controls. Platelets of bone marrow transplant recipients showed partly a significant, higher expression of surface bound fibrinogen, activated fibrinogen receptor, and glycoprotein Ib as compared to controls. P-selectin, a marker of platelet degranulation was significantly elevated after ADP-induced stimulation at all timepoints compared to controls. Only marginal differences were found for GP IIb/IIIa surface expression. The data point to an increased platelet activation state in bone marrow transplant recipients which might contribute to the thrombotic phenomena observed in these patients. PMID- 9753704 TI - Influence of intact left atrial appendage on hemodynamic parameters of isolated guinea pig heart. AB - BACKGROUND: In isolated working guinea pig heart preparations using the conventional technique of cannulating the left atrium via the atrial appendage, the resulting cardiac output is often insufficiently low (15-20 ml/min). This is a problem in ischemia reperfusion studies where small absolute differences can be responsible for large relative changes comparing pre- and postischemic values. In an attempt to increase cardiac output, the left atrium was left intact in isolated working guinea pig hearts. The two techniques were compared for hemodynamic parameters and their similarity to the physiological condition. METHODS: The left atrium was cannulated either via the orifices of the pulmonary veins or via an incision in the atrial appendage with its subsequent ligation around the cannula (n = 45-46/group). After 20 min of pressure-volume work cardiac output, heart rate and oxygen partial pressures were measured and myocardial oxygen consumption and cardiac efficiency were calculated. RESULTS: Cardiac output was higher in hearts with intact atrial appendage (64 +/- 2 ml/min) than in hearts with ligated atrial appendage (33 +/- 1 ml/min). Myocardial oxygen consumption (6.1 +/- 0.2 and 8.4 +/- 0.3 micromol/min, resp.) and cardiac efficiency (12.8 +/- 0.6% and 19.9 +/- 0.8%, resp.) were significantly higher in hearts with intact left atrial appendage. CONCLUSIONS: Isolated working guinea pig hearts with an intact left auricle exhibit higher values for important hemodynamic parameters compared to a preparation technique involving ligation of the left auricle. PMID- 9753705 TI - The analgesic effect of intravenous tenoxicam in symptomatic treatment of biliary colic: a comparison with hyoscine N-butylbromide. AB - This was conducted to evaluate the analgesic effect of intravenous tenoxicam (non steroidal anti-inflammatory drug) in the treatment of biliary colic pain and compared with spasmolytics. Thirtytwo patients (26 women, 6 men, mean age 47, range 38-55 years) with acute biliary colic were entered for study. They were allocated randomly to receive either tenoxicam 20 mg i.v. or hyoscine N butylbromide 20 mg i.v. The patients recorded their pain severity on 5 point scale. The results showed that tenoxicam caused significant pain relief in 10 out of 16 patients at 30 min (mean pain score decreased from 2.75 +/- 0.93 to 0.49 +/ 0.51, p < 0.05) and in other 4 patients at 60 min (mean pain score decreased to 0.58 + 5.7, p < 0.05). None of these patients developed acute cholecystitis or pain relapse over a period of 24 h follow up. With use of hyoscine N butylbromide, 7 out 16 patients had significant pain relief at 30 min (mean pain score decreased from 2.62 +/- 1.01 to 0.57 +/- 0.53, p < 0.05) and 3 other patients relieved at 60 min (mean pain score decreased to 0.66 +/- 0.57, p < 0.05). Four patients showed pain relapse within 24 h and needed pethidine-rescue treatment, two of them developed acute cholecystitis. Three out of 6 patients who showed no response to hyoscine N-butylbropmide and treated with 100 mg pethidine progressed to acute cholecystitis. We concluded that intravenous tenoxicam has rapid and prolong analgesic effects in the treatment of acute biliary colic as compared to hyoscine N-butylbroimde and it prevents the progression to acute cholecystitis. PMID- 9753706 TI - Oral health status of hospitalized children with cancer: a comparative study. AB - In this study, oral findings from children suffering from leukemia or other cancers, and hospitalized for treatments, are compared with findings from hospitalized patients from the Children s Surgery Department, without systemic or general illnesses. The aim of the study is to determine possible differences in the status of oral health between the hospitalized cancer-ward children and children hospitalized, but with good general health, in order to develop special prophylactic measures as required. The visual oral findings included the prevalence of caries (DMF/T, Decayed, Missed, Filled/Tooth), oral hygiene, severe periodontal diseases, records of defects of the oral mucosa, and information was collected on fluoridation. For both groups of children, the degree of treatment need of the remaining dentition was 63%. Caries-free dentition was found in 53.7% of the surgery patients and 40.7% of the cancer-ward children. Good oral hygiene was found in only 49% of the cases among the cancer-ward children, while among the children without systemic illness the figure climbed to 87% of the cases. Gingivitis among the children without general illness was only diagnosed in 7.4% of the cases, but in contrast, among the cancer-ward children the percentage of gingivitis (62.8% ) was significantly higher (p < 0.0001). The present study strongly suggests that for children with cancer and subject to aggressive therapy and/or long hospitalizations, beyond the general medical examination on hospital admission, a dental examination should also be instituted. PMID- 9753707 TI - HIV postexposure prophylaxis. German-Austrian recommendations. Deutsche AIDS Gesellschaft (DAIG) and Osterreichische AIDS- Gesellschaft (OAG). PMID- 9753708 TI - The sentinel node in breast cancer--a multicenter validation study. AB - BACKGROUND: Pilot studies indicate that probe-guided resection of radioactive sentinel nodes (the first nodes that receive drainage from tumors) can identify regional metastases in patients with breast cancer. To confirm this finding, we conducted a multicenter study of the method as used by 11 surgeons in a variety of practice settings. METHODS: We enrolled 443 patients with breast cancer. The technique involved the injection of 4 ml of technetium-99m sulfur colloid (1 mCi [37 MBq]) into the breast around the tumor or biopsy cavity. "Hot spots" representing underlying sentinel nodes were identified with a gamma probe. Sentinel nodes subjacent to hot spots were removed. All patients underwent a complete axillary lymphadenectomy. RESULTS: The overall rate of identification of hot spots was 93 percent (in 413 of 443 patients). The pathological status of the sentinel nodes was compared with that of the remaining axillary nodes. The accuracy of the sentinel nodes with respect to the positive or negative status of the axillary nodes was 97 percent (392 of 405); the specificity of the method was 100 percent, the positive predictive value was 100 percent, the negative predictive value was 96 percent (291 of 304), and the sensitivity was 89 percent (101 of 114). The sentinel nodes were outside the axilla in 8 percent of cases and outside of level 1 nodes in 11 percent of cases. Three percent of positive sentinel nodes were in nonaxillary locations. CONCLUSIONS: Biopsy of sentinel nodes can predict the presence or absence of axillary-node metastases in patients with breast cancer. However, the procedure can be technically challenging, and the success rate varies according to the surgeon and the characteristics of the patient. PMID- 9753709 TI - Cyclic administration of pamidronate in children with severe osteogenesis imperfecta. AB - BACKGROUND: Severe osteogenesis imperfecta is a disorder characterized by osteopenia, frequent fractures, progressive deformity, loss of mobility, and chronic bone pain. There is no effective therapy for the disorder. We assessed the effects of treatment with a bisphosphonate on bone resorption. METHODS: In an uncontrolled observational study involving 30 children who were 3 to 16 years old and had severe osteogenesis imperfecta, we administered pamidronate intravenously (mean [+/-SD] dose, 6.8+/-1.1 mg per kilogram of body weight per year) at 4-to-6 month intervals for 1.3 to 5.0 years. Clinical status, biochemical characteristics reflecting bone turnover, the bone mineral density of the lumbar spine, and radiologic changes were assessed regularly during treatment. RESULTS: Administration of pamidronate resulted in sustained reductions in serum alkaline phosphatase concentrations and in the urinary excretion of calcium and type I collagen N-telopeptide. There was a mean annualized increase of 41.9+/-29.0 percent in bone mineral density, and the deviation of bone mineral density from normal, as indicated by the z score, improved from -5.3+/-1.2 to -3.4+/-1.5. The cortical width of the metacarpals increased by 27+/-20.2 percent per year. The increases in the size of the vertebral bodies suggested that new bone had formed. The mean incidence of radiologically confirmed fractures decreased by 1.7 per year (P<0.001). Treatment with pamidronate did not alter the rate of fracture healing, the growth rate, or the appearance of the growth plates. Mobility and ambulation improved in 16 children and remained unchanged in the other 14. All the children reported substantial relief of chronic pain and fatigue. CONCLUSIONS: In children with severe osteogenesis imperfecta, cyclic administration of intravenous pamidronate improved clinical outcomes, reduced bone resorption, and increased bone density. PMID- 9753710 TI - Obesity associated with a mutation in a genetic regulator of adipocyte differentiation. AB - BACKGROUND: There is increasing evidence of genetic factors leading to obesity, but the exact genes involved have not been defined. Peroxisome-proliferator activated receptor gamma2 (PPARgamma2) is a transcription factor that has a key role in adipocyte differentiation, and therefore mutations of the gene for this factor might predispose people to obesity. METHODS: We studied 358 unrelated German subjects, including 121 obese subjects (defined as those with a body-mass index [the weight in kilograms divided by the square of the height in meters] of more than 29). We evaluated these subjects for mutations in the gene for PPARgamma2 at or near a site of serine phosphorylation at position 114 that negatively regulates the transcriptional activity of the protein, using a polymerase-chain-reaction-based assay coupled with specific endonuclease digestion. The activity of the mutation identified was analyzed by retroviral transfection and overexpression in murine fibroblasts. RESULTS: Four of the 121 obese subjects had a missense mutation in the gene for PPARgamma2 that resulted in the conversion of proline to glutamine at position 115, as compared with none of the 237 subjects of normal weight (P=0.01). All the subjects with the mutant allele were markedly obese, with body-mass-index values ranging from 37.9 to 47.3, as compared with a mean of 33.6 in the other obese subjects. Overexpression of the mutant gene in murine fibroblasts led to the production of a protein in which the phosphorylation of serine at position 114 was defective, as well as to accelerated differentiation of the cells into adipocytes and greater cellular accumulation of triglyceride than with the wild-type PPARgamma2. These effects were similar to those of an in vitro mutation created directly at the Ser114 phosphorylation site. CONCLUSIONS: A Pro115Gln mutation in PPARgamma2 accelerates the differentiation of adipocytes and may cause obesity. PMID- 9753711 TI - Influence of the genotype on the clinical course of the long-QT syndrome. International Long-QT Syndrome Registry Research Group. AB - BACKGROUND: The congenital long-QT syndrome, caused by mutations in cardiac potassium-channel genes (KVLQT1 at the LQT1 locus and HERG at the LQT2 locus) and the sodium-channel gene (SCN5A at the LQT3 locus), has distinct repolarization patterns on electrocardiography, but it is not known whether the genotype influences the clinical course of the disease. METHODS: We determined the genotypes of 541 of 1378 members of 38 families enrolled in the International Long-QT Syndrome Registry: 112 had mutations at the LQT1 locus, 72 had mutations at the LQT2 locus, and 62 had mutations at the LQT3 locus. We determined the cumulative probability and lethality of cardiac events (syncope, aborted cardiac arrest, or sudden death) occurring from birth through the age of 40 years according to genotype in the 246 gene carriers and in all 1378 members of the families studied. RESULTS: The frequency of cardiac events was higher among subjects with mutations at the LQT1 locus (63 percent) or the LQT2 locus (46 percent) than among subjects with mutations at the LQT3 locus (18 percent) (P<0.001 for the comparison of all three groups). In a multivariate Cox analysis, the genotype and the QT interval corrected for heart rate were significant independent predictors of a first cardiac event. The cumulative mortality through the age of 40 among members of the three groups of families studied was similar; however, the likelihood of dying during a cardiac event was significantly higher (P<0.001) among families with mutations at the LQT3 locus (20 percent) than among those with mutations at the LQT1 locus (4 percent) or the LQT2 locus (4 percent). CONCLUSIONS: The genotype of the long-QT syndrome influences the clinical course. The risk of cardiac events is significantly higher among subjects with mutations at the LQT1 or LQT2 locus than among those with mutations at the LQT3 locus. Although cumulative mortality is similar regardless of the genotype, the percentage of cardiac events that are lethal is significantly higher in families with mutations at the LQT3 locus. PMID- 9753712 TI - Images in clinical medicine. Blue sclerae in osteogenesis imperfecta. PMID- 9753713 TI - Attitudes of patients with amyotrophic lateral sclerosis and their care givers toward assisted suicide. AB - BACKGROUND AND METHODS: Amyotrophic lateral sclerosis (ALS) is a neuromuscular disease that causes gradual paralysis, respiratory failure, and death, usually within three to five years after it has been diagnosed. Between 1995 and 1997, we surveyed patients with this disease in Oregon and Washington, as well as their family care givers, in order to determine their attitudes toward assisted suicide. Patients were considered to be willing to contemplate assisted suicide if they agreed with the statement, "Under some circumstances I would consider taking a prescription for a medicine whose sole purpose was to end my life," and disagreed with the statement, "I would never request or take a prescription for a medication whose sole purpose was to end my life." The Oregon Death with Dignity Act, which legalized physician-assisted suicide, was approved by Oregon voters in 1994 but did not go into effect until October 1997, after data collection for this study had been completed. RESULTS: Of 140 eligible persons with ALS, 100 (71 percent) agreed to participate in the study, as did 91 family care givers. The mean age of the patients with ALS was 54 years; the mean duration of illness since the diagnosis was 2.8 years. Fifty-six patients (56 percent) said they would consider assisted suicide, and 44 of the 56 agreed with the statement, "If physician-assisted suicide were legal, I would request a lethal prescription from a physician." One patient would have taken the medication immediately, and 36 would have kept it for future use. As compared with the patients who were opposed to assisted suicide, those who would consider it were more likely to be men, had a higher level of education, were less likely to be religious, had higher scores for hopelessness, and rated their quality of life as lower. In 66 of 91 instances (73 percent), care givers and patients had the same attitude toward assisted suicide. CONCLUSIONS: In Oregon and Washington, a majority of persons with ALS whom we surveyed would consider assisted suicide. Many would request a prescription for a lethal dose of medication well before they intended to use it. PMID- 9753714 TI - Treatment of breast cancer. PMID- 9753715 TI - Osteogenesis imperfecta--managing brittle bones. PMID- 9753716 TI - Assisted suicide and alternatives in amyotrophic lateral sclerosis. PMID- 9753717 TI - Sentinel-lymph-node biopsy for breast cancer--not yet the standard of care. PMID- 9753718 TI - Merian Frederick's story. PMID- 9753719 TI - Does SINE evolution preclude Alu function? AB - The evolution, mobility and deleterious genetic effects of human Alus are fairly well understood. The complexity of regulated transcriptional expression of Alus is becoming apparent and insight into the mechanism of retrotransposition is emerging. Unresolved questions concern why mobile, highly repetitive short interspersed elements (SINEs) have been tolerated throughout evolution and why and how families of such sequences are periodically replaced. Either certain SINEs are more successful genomic parasites or positive selection drives their relative success and genomic maintenance. A complete understanding of the evolutionary dynamics and significance of SINEs requires determining whether or not they have a function(s). Recent evidence suggests two possibilities, one concerning DNA and the other RNA. Dispersed Alus exhibit remarkable tissue specific differences in the level of their 5-methylcytosine content. Differences in Alu methylation in the male and female germlines suggest that Alu DNA may be involved in either the unique chromatin organization of sperm or signaling events in the early embryo. Alu RNA is increased by cellular insults and stimulates protein synthesis by inhibiting PKR, the eIF2 kinase that is regulated by double stranded RNA. PKR serves other roles potentially linking Alu RNA to a variety of vital cell functions. Since Alus have appeared only recently within the primate lineage, this proposal provokes the challenging question of how Alu RNA could have possibly assumed a significant role in cell physiology. PMID- 9753720 TI - Anomeric inversion (from beta to alpha) in methylphosphonate oligonucleosides enhances their affinity for DNA and RNA. AB - Here we report that the poor binding of methylphosphonate oligodeoxynucleosides (MP-ODNs) to their nucleic acid targets can be improved by additional inversion of the anomeric configuration (from beta to alpha) in the sugar moieties to give a new class of analogs, MP alpha-oligonucleosides. MP alpha-dT12and MP 5' alpha d(TCTTAACCCACA) 3' were synthesized and their ability to form hybrids with complementary single stranded (ss)DNA and ssRNA, as well as with double stranded (ds)DNA, was evaluated. The thermal stability of hybrids formed with MP alpha analogs was compared with the affinity of phosphodiester (PO) and phosphorothioate (PS) beta- and alpha-oligomers for their targets. Non-ionic MP alpha-oligonucleosides bound to their complementary DNA and RNA strands more tightly than their homologues with natural beta-anomeric configuration did. With DNA target, MP alpha-oligomers formed duplexes more stable than the corresponding natural PO beta-oligomer did. MP alpha-heteropolymer hybridized to RNA target better than PS beta-oligonucleotide did but the hybrid was less stable (DeltaTm 0.5 degrees C per mod.) than the hybrid formed with the natural PO beta-oligomer. Only MP alpha-dT12 bound to dsDNA target at low salt concentration (0.1 M NaCl). PMID- 9753721 TI - The hypotrichous ciliate Euplotes octocarinatus has only one type of tRNACys with GCA anticodon encoded on a single macronuclear DNA molecule. AB - Deviations from the universal genetic code have evolved independently several times in ciliated protozoa. Thus, in some species UAA and UAG are no longer used as termination codons, but are read as glutamine, whereas in the genus Euplotes , UGA is translated as cysteine. We have investigated the nature of the tRNACys isoacceptor responsible for decoding UGA in Euplotes cells. Southern hybridization analyses indicated that a single DNA molecule of 630 bp encoding tRNACys exists in the macronucleus of Euplotes octocarinatus . Cloning and sequencing of this fragment revealed that it contains only one copy of a tRNACys gene, which codes for a normal tRNACys with GCA anticodon. This is the first report of the characterization of a tRNA gene in any hypotrichous ciliate. It contains putative signals for initiation and termination of transcription by RNA polymerase III and can be transcribed efficiently in vitro in HeLa cell nuclear extract. Intensive studies on the DNA and tRNA level involving PCR analyses have not disclosed the existence of any tRNA Cys isoacceptor with UCA or ICA anticodons. Translation of the UGA codon by tRNA sub GCA sup Cys necessitates a G:A mispairing in the first anticodon position. We discuss a number of aspects which might contribute to the finding that a near-cognate tRNA isoacceptor efficiently translates the UGA stop codon. PMID- 9753722 TI - Position effect takes precedence over target sequence in determination of adenine methylation patterns in the nuclear genome of a eukaryote, Tetrahymena thermophila. AB - Approximately 0.8% of the adenine residues in the macronuclear DNA of the ciliated protozoan Tetrahymena thermophila are modified to N 6-methyladenine. DNA methylation is site specific and the pattern of methylation is constant between clonal cell lines. In vivo, modification of adenine residues appears to occur exclusively in the sequence 5'-NAT-3', but no consensus sequence for modified sites has been found. In this study, DNA fragments containing a site that is uniformly methylated on the 50 copies of the macronuclear chromosome were cloned into the extrachromosomal rDNA. In the novel location on the rDNA minichromosome, the site was unmethylated. The result was the same whether the sequences were introduced in a methylated or unmethylated state and regardless of the orientation of the sequence with respect to the origin of DNA replication. The data show that sequence is insufficient to account for site-specific methylation in Tetrahymena and argue that other factors determine the pattern of DNA methylation. PMID- 9753723 TI - Helix 2 of the paired domain plays a key role in the regulation of DNA-binding by the Pax-3 homeodomain. AB - Pax3 contains two structurally independent DNA-binding domains, a paired domain (PD) and a homeodomain (HD). Biochemical and mutagenesis studies have shown that both domains are functionally interdependent. In particular, it has been shown that the PD can regulate the DNA-binding specificity and dimerization potential of the HD. To delineate Pax3 protein segments that are involved in the regulation of HD DNA-binding, a series of chimeric proteins were created in which the HD and linker region were gradually replaced with corresponding sequences from a heterologous HD protein, Phox. Characterization of chimeric proteins by electrophoretic mobility shift analysis (EMSA) suggests that a portion of the linker region contributes to the functional interaction between the PD and HD. In addition, stepwise removal of sequences from the Pax3 PD was used to define regions within this domain that are involved in the regulation of HD DNA-binding. EMSA of these proteins in the context of the chimeric Pax3/Phox backbone provided two key findings: (i) the C-terminal subdomain of the PD does not play a major role in the regulation of HD DNA-binding and (ii) the N-terminal subdomain and, in particular, the second alpha-helix are essential for modulation of HD DNA binding. Significantly, deletion of helix 2 was found to be sufficient to uncouple regulation of HD DNA-binding by the PD. PMID- 9753724 TI - The mutations induced by oxidatively damaged nucleotides, 5-formyl-dUTP and 5 hydroxy-dCTP,in Escherichia coli. AB - The mutational properties of 5-formyl-2'-deoxyuridine 5'-triphosphate (5-CHO dUTP) and 5-hydroxy-2'-deoxycytidine 5'-triphosphate (5-OH-dCTP), the major oxidatively damaged pyrimidine nucleotides derived from dTTP and dCTP, respectively, were analyzed by an in vivo assay. 5-CHO-dUTP and 5-OH-dCTP were directly incorporated into Escherichia coli , and their mutagenicities were evaluated by the chromosomal lacI forward mutation assay. The mutation frequencies increased, depending on the dose of these damaged nucleotides, indicating that these nucleotides were incorporated into E.coli and acted as mutagens in vivo . The mutagenicities of 5-CHO-dUTP and 5-OH-dCTP were comparable to that of 8-hydroxy-2'-deoxyguanosine 5'-triphosphate, a major form of dGTP oxidative damage. 5-CHO-dUTP induced G.C to A.T, A.T to G.C and G.C to T.A mutations, and 5-OH-dCTP elicited G.C to A.T, A.T to C.G and G.C to T.A mutations. PMID- 9753725 TI - The yeast FBP1 poly(A) signal functions in both orientations and overlaps with a gene promoter. AB - This report provides an analysis of a region of chromosome XII in which the FBP1 and YLR376c genes transcribe in the same direction. Our investigation indicates that the Saccharomyces cerevisiae FBP1 gene contains strong signals for polyadenylation and transcription termination in both orientations in vivo . A (TA)14 element plays a major role in directing polyadenylation in both orientations. While this region has four nonoverlapping copies of a TATATA hexanucleotide, which is a very potent polyadenylation efficiency element in yeast, it alone is not sufficient for full activation in the reverse orientation of a cluster of downstream poly(A) sites, and an additional upstream sequence is required. The putative RNA hairpin formed from the (TA)14 element is not involved in 3'-end formation. Surprisingly, deletion of the entire (TA)14 stretch affects transcription termination in the reverse orientation, in contrast to our previous results with the forward orientation, indicating that the transcription termination element operating in the reverse orientation has very different sequence requirements. Promoter elements for the YLR376c gene overlap with the signal for FBP1 3'-end formation. To our knowledge, this is the first time that overlapping of both types of regulatory signals has been found in two adjacent yeast genes. PMID- 9753726 TI - Mutation detection using a novel plant endonuclease. AB - We have discovered a useful new reagent for mutation detection, a novel nuclease CEL I from celery. It is specific for DNA distortions and mismatches from pH 6 to 9. Incision is on the 3'-side of the mismatch site in one of the two DNA strands in a heteroduplex. CEL I-like nucleases are found in many plants. We report here that a simple method of enzyme mutation detection using CEL I can efficiently identify mutations and polymorphisms. To illustrate the efficacy of this approach, the exons of the BRCA1 gene were amplified by PCR using primers 5' labeled with fluorescent dyes of two colors. The PCR products were annealed to form heteroduplexes and subjected to CEL I incision. In GeneScan analyses with a PE Applied Biosystems automated DNA sequencer, two independent incision events, one in each strand, produce truncated fragments of two colors that complement each other to confirm the position of the mismatch. CEL I can detect 100% of the sequence variants present, including deletions, insertions and missense alterations. Our results indicate that CEL I mutation detection is a highly sensitive method for detecting both polymorphisms and disease-causing mutations in DNA fragments as long as 1120 bp in length. PMID- 9753727 TI - High resolution mapping DNAs by R-loop atomic force microscopy. AB - R-loops formed by short RNA transcripts have been imaged by atomic force microscopy (AFM) at a constant force in the height mode. The technique was applied to mapping the human endogenous retrovirus K10 family (HERV-K10) long terminal repeats (LTR) within individual plasmids and cosmids. RNA probes specific for the U3 (384 nt) and U5 (375 nt) LTR regions separated by a span of 200 bp were used for R-loop formation with LTRs located within plasmid (3.8 kb) or cosmid ( approximately 40 kb) DNAs. R-loops stabilized by glyoxal treatment and adsorbed onto the mica surface in the presence of magnesium ions looked like looped out segments of RNA:DNA hybrids. The total yield of R-loops was usually approximately 95%. The RNA:DNA hybrids were found to be 12-15% shorter than the corresponding DNA:DNA duplex. The two regions of the LTR could be easily discerned in the AFM images as clearly separated loops. R-loop positions determined on cosmids by AFM were accurate to approximately 0.5% of the cosmid length. This technique might be easily adapted for mapping various sequences such as gene exons or regulatory regions and for detecting insertions, deletions and rearrangements that cause human genetic diseases. PMID- 9753728 TI - Nuclear and mitochondrial splice forms of human uracil-DNA glycosylase contain a complex nuclear localisation signal and a strong classical mitochondrial localisation signal, respectively. AB - Nuclear (UNG2) and mitochondrial (UNG1) forms of human uracil-DNA glycosylase are both encoded by the UNG gene but have different N-terminal sequences. We have expressed fusion constructs of truncated or site-mutated UNG cDNAs and green fluorescent protein cDNA and studied subcellular sorting. The unique 44 N terminal amino acids in UNG2 are required, but not sufficient, for complete sorting to nuclei. In this part the motif R17K18R19is essential for sorting. The complete nuclear localization signal (NLS) in addition requires residues common to UNG2 and UNG1 within the 151 N-terminal residues. Replacement of certain basic residues within this region changed the pattern of subnuclear distribution of UNG2. The 35 unique N-terminal residues in UNG1 constitute a strong and complete mitochondrial localization signal (MLS) which when placed at the N-terminus of UNG2 overrides the NLS. Residues 11-28 in UNG1 have the potential of forming an amphiphilic helix typical of MLSs and residues 1-28 are essential and sufficient for mitochondrial import. These results demonstrate that UNG1 contains a classical and very strong MLS, whereas UNG2 contains an unusually long and complex NLS, as well as subnuclear targeting signals in the region common to UNG2 and UNG1. PMID- 9753729 TI - Mutational analysis of Chlorella virus DNA ligase: catalytic roles of domain I and motif VI. AB - A conserved catalytic core of the ATP-dependent DNA ligases is composed of an N terminal domain (domain 1, containing nucleotidyl transferase motifs I, III, IIIa and IV) and a C-terminal domain (domain 2, containing motif VI) with an intervening cleft. Motif V links the two structural domains. Deletion analysis of the 298 amino acid Chlorella virus DNA ligase indicates that motif VI plays a critical role in the reaction of ligase with ATP to form ligase-adenylate, but is dispensable for the two subsequent steps in the ligation pathway; DNA-adenylate formation and strand closure. We find that formation of a phosphodiester at a pre adenylated nick is subject to a rate limiting step that does not apply during the sealing of nicked DNA by ligase-adenylate. This step, presumably conformational, is accelerated or circumvented by deleting five amino acids of motif VI. The motif I lysine nucleophile (Lys27) is not required for strand closure by wild type ligase, but this residue enhances the closure rate by a factor of 16 when motif VI is truncated. We find that a more extensively truncated ligase consisting of only N-terminal domain 1 and motif V is inert in ligase--adenylate formation, but competent to catalyze strand closure at a pre-adenylated nick. These results suggest that different enzymic catalysts facilitate the three steps of the DNA ligase reaction. PMID- 9753730 TI - Regulated processive transcription of chromatin by T7 RNA polymerase in Trypanosoma brucei. AB - Inability of T7 RNA polymerase to processively transcribe higher eukaryotic chromatin is interpreted as a correlate of its reported inhibition by nucleosomes on reconstituted templates in vitro . We used chromosomally integrated reporter cassettes to examine features of T7 transcription in a lower eukaryotic system. Luciferase reporters were targeted to rDNA in transgenic Trypanosoma brucei stably expressing the phage polymerase. Because trypanosome mRNAs are capped by RNA splicing in trans , T7 transcription could be gauged by luciferase activity. In contrast to findings from higher eukaryotes, T7 transcription is vigorous and processive on chromatin templates in T.brucei , surpassing levels achieved with endogenous promoters, including those recruiting RNA polymerase I. This may be a reflection of intrinsic differences in chromatin structure between differently evolved eukaryotes or of an integration site that is exceptionally permissive for T7 transcription due to a local accessible chromatin conformation. T7 transcription could be manipulated to achieve different levels of constitutive expression, through the use of promoter mutations. Moreover, T7 initiation could be regulated by the prokaryotic Tet repressor and elongation halted by T7 terminator sequences. We have exploited these features to construct a robust inducible expression system, whose utility potentially extends to other trans splicing organisms. PMID- 9753731 TI - Conserved cis- and trans-acting determinants for replication initiation and regulation of replication fork movement in tetrahymenid species. AB - The rDNA minichromosomes of Tetrahymena thermophila and Tetrahymena pyriformis share a high degree of sequence similarity and structural organization. The T.thermophila 5' non-transcribed spacer (5' NTS) is sufficient for replication and contains three repeated sequence elements that are conserved in T.pyriformis , including type I elements, the only known determinant for replication control. To assess the role of conserved sequences in replication control, structural and functional studies were performed on T.pyriformis rDNA. Similar to T.thermophila , replication initiates exclusively in the 5' NTS, localizing to a 900 bp segment. Elongating replication forks arrest transiently at one site which bears strong similarity to a tripartite sequence element present at fork arrest sites in T.thermophila rDNA. An in vitro type I element binding activity indistinguishable from the T.thermophila protein, ssA-TIBF, was detected in T.pyriformis extracts. The respective TIBF proteins bind with comparable affinity to type I elements from both species, suggesting that in vivo recognition could cross species boundaries. Despite these similarities, the T.pyriformis 5' NTS failed to support replication in transformed T.thermophila cells, suggesting a more complex genetic organization than previously realized. PMID- 9753732 TI - The LAZ3(BCL-6) oncoprotein recruits a SMRT/mSIN3A/histone deacetylase containing complex to mediate transcriptional repression. AB - Recent works demonstrated that some transcriptional repressors recruit histone deacetylases (HDACs) either through direct interaction, or as a member of a multisubunit repressing complex containing other components referred to as corepressors. For instance, the bHLH-Zip transcriptional repressors MAD/MXI recruit HDACs together with the mSIN3 corepressors, whereas unliganded nuclear receptors contact another corepressor, SMRT (or its relative N-CoR), which, in turn, associates with both mSIN3 and HDACs to form the repressor complex. Recently, we reported that SMRT also directly associates with LAZ3(BCL-6), a POZ/Zn finger transcriptional repressor involvedin the pathogenesis of non Hodgkin lymphomas. However, whether LAZ3 recruits the HDACs-containing repression complex is currently unknown. We report here that LAZ3 associates with corepressor mSIN3A both in vivo and in vitro , and found that a central region, which harbours autonomous repression activity, is mainly responsible for this interaction. Conversely, the N-terminal half of mSIN3A is both necessary and sufficient to bind LAZ3. Moreover, we show that LAZ3 also interacts with an HDAC (HDAC-1) through its POZ domain in vitro while the immunoprecipitation of LAZ3 results in the coretention of an endogenous HDAC activity in vivo . Finally, inhibitors of HDACs significantly reduce the LAZ3-mediated repression. Taken together, we conclude that LAZ3 recruits a repressing complex containing SMRT, mSIN3A and a HDAC, and that its full repressing potential on transcription requires HDACs activity. Our results identify HDACs as molecular targets of LAZ3 oncogene and further strengthen the connection between aberrant chromatin acetylation and human cancers. PMID- 9753733 TI - Creation of genetic information by DNA polymerase of the archaeon Thermococcus litoralis: influences of temperature and ionic strength. AB - DNA polymerase of the archaeon Thermococcus litoralis can synthesize a long stretch of linear double-stranded DNA in the complete absence of added primer and template DNAs. This finding suggests that genetic information can potentially be created by protein. We report here the effects of temperature, ionic strength and pH on this ab initio DNA synthesis by the protein in vitro . When the temperature of the reaction was changed, the sequence of the product DNA changed markedly. For instance, the reaction products were (TAAT) n at 69 degrees C, (TATCCGGA) n at 84 degrees C and (TATCGCGATAGCGATCGC) n at 89 degrees C. The ionic strength of the reaction condition also affected the sequence: it was (TATCTAGA) n with 0 mM KCl, (TATATACG) n with 50 mM KCl and (TATAGTTATAAC) n with 100 mM KCl at 74 degrees C. When the pH of the reaction condition was changed from 6.8 to 10.8, the size of the product DNA decreased, but its sequence did not. These results demonstrate that DNA synthesized ab initio by DNA polymerase of T.litoralis is markedly influenced by the reaction conditions. The results also suggest that genetic information that might have been created by protein on the early earth is strongly influenced by environmental factors. PMID- 9753734 TI - Creation of genetic information by DNA polymerase of the thermophilic bacterium Thermus thermophilus. AB - Genetic information encoded in a template of a genome is replicated in a complementary way by DNA polymerase or RNA polymerase with high fidelity; no creation of information occurs in this reaction unless an error occurs. We report here that DNA polymerase of the thermophilic bacterium Thermus thermophilus can synthesize up to 200 kb linear double-stranded DNA in vitro in the complete absence of added primer and template DNAs, indicating that genetic information is actively created by protein. This ab initio DNA synthesis occurs at 74 degrees C and requires magnesium ion. There is a lag time of approximately 1 h and then the reaction proceeds linearly. The synthesized DNAs have a variety of sequences; they are mostly tandem repetitive sequences, e.g. (CATGTATA) n , (TGTATGTATACATACATA) n and (TATACGTA) n . Some degenerate sequences of these basic repeat units are also found. The similar repetitive sequences are found in many natural genes. These results, together with similar results found using DNA polymerase of archaeon Thermococcus litoralis , suggest that creative, non replicative synthesis of DNA by protein was a driving force for diversification of genetic information at a certain stage of the evolution of life on the early earth. PMID- 9753735 TI - Mutational analysis of a function of xeroderma pigmentosum group A (XPA) protein in strand-specific DNA repair. AB - To analyze the function of the xeroderma pigmentosum group A (XPA) protein in strand-specific DNA repair, we examined repair of UV-induced cyclobutane pyrimidine dimer (CPD) in transcribed and non-transcribed strands of the dihydrofolate reductase gene of xeroderma pigmentosum group A (XP-A) cell line (XP12ROSV) which was transfected with various types of mutant XPA cDNA. The transfectant overexpressing mutant XPA with a defect in the interaction with either ERCC1, replication protein A (RPA), or general transcription factor TFIIH, showed more or less decreased repair of CPD in each strand in parallel, while in the transfectant overexpressing R207G (Arg207to Gly) mutant XPA derived from XP129, a UV-resistant XP12ROSV revertant, the rate of CPD repair was almost normal in each strand. We also examined the dose responses of the XPA protein on CPD repair in each strand by the modulation of the expression levels of wild-type or R207G mutant XPA using an inducible expression system, LacSwitchtrade mark promoter. There were good correlations between the rate of CPD repair in each strand and the amount of XPA protein produced in these Lac cells. Our results indicate that the XPA protein is equally important for the CPD repair in both transcribed and non-transcribed strands and that the R207G mutation found in XP129 may not be responsible for a selective defect in CPD repair in the non transcribed strand in XP129. PMID- 9753736 TI - Escherichia coli MutY protein has a guanine-DNA glycosylase that acts on 7,8 dihydro-8-oxoguanine:guanine mispair to prevent spontaneous G:C-->C:G transversions. AB - Low rates of spontaneous G:C-->C:G transversions would be achieved not only by the correction of base mismatches during DNA replication but also by the prevention and removal of oxidative base damage in DNA. Escherichia coli must have several pathways to repair such mismatches and DNA modifications. In this study, we attempted to identify mutator loci leading to G:C-->C:G transversions in E.coli. The strain CC103 carrying a specific mutation in lacZ was mutagenized by random miniTn 10 insertion mutagenesis. In this strain, only the G:C-->C:G change can revert the glutamic acid at codon 461, which is essential for sufficient beta-galactosidase activity to allow growth on lactose. Mutator strains were detected as colonies with significantly increased rates of papillae formation on glucose minimal plates containing P-Gal and X-Gal. We screened approximately 40 000 colonies and selected several mutator strains. The strain GC39 showed the highest mutation rate to Lac+. The gene responsible for the mutator phenotypes, mut39 , was mapped at around 67 min on the E.coli chromosome. The sequencing of the miniTn 10 -flanking DNA region revealed that the mut39 was identical to the mutY gene of E.coli. The plasmid carrying the mutY + gene reduced spontaneous G:C-->T:A and G:C-->C:G mutations in both mutY and mut39 strains. Purified MutY protein bound to the oligonucleotides containing 7,8 dihydro-8-oxo-guanine (8-oxoG):G and 8-oxoG:A. Furthermore, we found that the MutY protein had a DNA glycosylase activity which removes unmodified guanine from the 8-oxoG:G mispair. These results demonstrate that the MutY protein prevents the generation of G:C-->C:G transversions by removing guanine from the 8-oxoG:G mispair in E.coli. PMID- 9753737 TI - Regulation of poly(A) site choice of several yeast mRNAs. AB - Several yeast genes produce multiple transcripts with different 3'-ends. Of these, four genes are known to produce truncated transcripts that end within the coding sequence of longer transcripts: CBP1 , AEP2 / ATP13 , RNA14 and SIR1 . It has been shown that the level of the truncated CBP1 transcript increases during the switch to respiratory growth while that of the full-length transcript decreases. To determine whether this phenomenon is unique to CBP1 , northern analysis was used to determine whether the levels of other truncated transcripts are regulated similarly by carbon source. The levels of the shortest transcripts of AEP2 / ATP13 and RNA14 increased during respiration while the shortest SIR1 transcript remained constant. However, two longer SIR1 transcripts were regulated reciprocally by carbon source. Mapping the 3'-ends of each transcript by sequencing partial cDNA clones revealed multiple 3'-ends for each transcript. Examination of the sequences surrounding the 3'-ends of the induced transcripts failed to identify a consensus sequence but did reveal weak putative 3'-end formation signals in all of the transcripts. Similarly, no consensus sequence was found when the sequences surrounding the 3'-ends of the longest transcripts were compared, but again weak putative 3'-end formation signals were identified. These data are suggestive of carbon source regulation of alternative poly(A) site choice in yeast. PMID- 9753738 TI - Mapping of a protein-RNA kissing hairpin interface: Rom and Tar-Tar*. AB - An RNA 'kissing' complex is formed by the association of two hairpins via base pairing of their complementary loops. This sense-antisense RNA motif is used in the regulation of many cellular processes, including Escherichia coli ColE1 plasmid copy number. The RNA one modulator protein (Rom) acts as a co-regulator of ColE1 plasmid copy number by binding to the kissing hairpins and stabilizing their interaction. We have used heteronuclear two-dimensional NMR spectroscopy to map the interface between Rom and a kissing complex formed by the loop of the trans -activation response (Tar) element of immunodeficiency virus 1 (HIV-1) and its complement. The protein binding interface was obtained from changes in amide proton signals of uniformly 15N-labeled Rom with increasing concentrations of unlabeled Tar-Tar*. Similarly, the RNA-binding interface was obtained from changes in imino proton signals of uniformly 15N-labeled Tar with increasing concentrations of unlabeled Rom. Our results are in agreement with previous mutagenesis studies and provide additional information on Rom residues involved in RNA binding. The kissing hairpin interface with Rom leads to a model in which the protein contacts the minor groove of the loop-loop helix and, to a lesser extent, the major groove of the stems. PMID- 9753739 TI - Intercalated cytosine motif and novel adenine clusters in the crystal structure of the Tetrahymena telomere. AB - The cytosine-rich strand of the Tetrahymena telomere consists of multiple repeats of sequence d(AACCCC). We have solved the crystal structure of the crystalline repeat sequence at 2.5 A resolution. The adenines form two different and previously unknown clusters (A clusters) in orthogonal directions with their counterparts from other strands, each containing a total of eight adenines. The clusters appear to be stable aggregates held together by base stacking and three different base-pairing modes. Two different types of cytosine tetraplexes are found in the crystal. Each four-stranded complex is composed of two intercalated parallel-stranded duplexes pointing in opposite directions, with hemiprotonated cytosine-cytosine (C.C+) base pairs. The outermost C.C+base pairs are from the 5' end of each strand in one cytosine tetraplex and from the 3'-end of each strand in the other. The A clusters and the cytosine tetraplexes form two alternating stacking patterns, creating continuous base stacking in two perpendicular directions along the x - and z -axes. The adenine clusters could be organizational motifs for macromolecular RNA. PMID- 9753740 TI - Solution structure of DAPI selectively bound in the minor groove of a DNA T.T mismatch-containing site: NMR and molecular dynamics studies. AB - The solution structure of the complex between 4', 6-diamidino-2-phenylindole (DAPI) and DNA oligomer [d(GCGATTCGC)]2, containing a central T.T mismatch, has been characterized by combined use of proton one- and two-dimensional NMR spectroscopy, molecular mechanics and molecular dynamics computations including relaxation matrix refinement. The results show that the DAPI molecule binds in the minor groove of the central region 5'-ATT-3' of the DNA oligomer, which predominantly adopts a duplex structure with a global right-handed B-like conformation. In the final models of the complex, the DAPI molecule is located nearly isohelical with its NH indole proton oriented towards the DNA helix axis and forming a bifurcated hydrogen bond with the carbonyl O2 groups of a mismatched T5 and the T6 residue of the opposite strand. Mismatched thymines adopt a wobble base pair conformation and are found stacked between the flanking base pairs, inducing only minor local conformational changes in global duplex structure. In addition, no other binding mechanisms were observed, showing that minor groove binding of DAPI to the mismatch-containing site is favoured in comparison with any other previously reported interaction with G.C sequences. PMID- 9753741 TI - The two homeodomains of the ZmHox2a gene from maize originated as an internal gene duplication and have evolved different target site specificities. AB - The maize ZmHox2a gene encodes two homeodomains which originated by a 699 bp duplication within an ancestral precursor. The sequences of the two ZmHox2a homeodomains are highly diverged in the N-terminal arm, while residues in the helical part have mostly been conserved. We show here that both ZmHox2a homeodomains are functional DNA-binding motifs but exhibit different target site specificities. CASTing experiments reveal a TCCT motif recognized by HD1 but a GATC tetranucleotide as the recognition sequence of HD2. Mutation of the central nucleotides in both tetranucleotide core motifs abolishes DNA binding. A domain swap experiment indicates that target site specificity is achieved in a combinatorial manner by the contributions of the diverged N-terminal arms together with the slightly different recognition helices. Computer modelling suggests that K47 and H54 in the recognition helices preferentially contact the bases at the 3'-terminus of the tetranucleotide target sequences. PMID- 9753742 TI - Identification and hydropathic characterization of structural features affecting sequence specificity for doxorubicin intercalation into DNA double-stranded polynucleotides. AB - The computer molecular modeling program HINT (Hydropathic INTeractions), an empirical hydropathic force field function that includes hydrogen bonding, coulombic and hydrophobic terms, was used to study sequence-selective doxorubicin binding/intercalation in the 64 unique CAxy, CGxy, TAxy, TGxy base pair quartet combinations. The CAAT quartet sequence is shown to have the highest binding score of the 64 combinations. Of the two regularly alternating polynucleotides, d(CGCGCG)2and d(TATATA)2, the HINT calculated binding scores reveal doxorubicin binds preferentially to d(TATATA)2. Although interactions of the chromophore with the DNA base pairs defining the intercalation site [I-1] [I+1] and the neighboring [I+2] base pair are predominant, the results obtained with HINT indicate that the base pair [I+3] contributes significantly to the sequence selectivity of doxorubicin by providing an additional hydrogen bonding opportunity for the N3' ammonium of the daunosamine sugar moiety in approximately 25% of the sequences. This observation, that interactions involving a base pair [I+3] distal to the intercalation site play a significant role in stabilizing/destabilizing the intercalation of doxorubicin into the various DNA sequences, has not been previously reported. In general terms, this work shows that molecular modeling and careful analysis of molecular interactions can have a significant role in designing and evaluating nucleotides and antineoplastic agents. PMID- 9753743 TI - A structure-specific DNA endonuclease is enriched in kinetoplasts purified from Crithidia fasciculata. AB - The mitochondrial DNA (kinetoplast DNA) of the trypanosomatid Crithidia fasciculata consists of minicircles and maxicircles topologically interlocked in a single network per cell. Individual minicircles replicate unidirectionally from either of two replication origins located 180 degrees apart on the minicircle DNA. Initiation of minicircle leading-strand synthesis involves the synthesis of an RNA primer which is removed in the last stage of replication. We report here the purification to near homogeneity of a structure-specific DNA endo-nuclease based on the RNase H activity of the enzyme on a poly(rA).poly(dT) substrate. RNase H activity gel analysis of whole cell and kinetoplast extracts shows that the enzyme is enriched in kinetoplast fractions. The DNA endonuclease activity of the enzyme is specific for DNA primers annealed to a template strand and requires an unannealed 5' tail. The enzyme cleaves 3' of the first base paired nucleotide releasing the intact tail. The purified enzyme migrates as a 32 kDa protein on SDS gels and has a Stoke's radius of 21.5 A and a sedimentation coefficient of 3.7 s, indicating that the protein is a monomer in solution with a native molecular mass of 32.4 kDa. These results suggest that the enzyme may be involved in RNA primer removal during minicircle replication. PMID- 9753744 TI - Molecular cloning and expression of the mouse translation initiation factor eIF 1A. AB - Prior to determining the molecular basis for the transient increase in expression of eIF-1A during the 2-cell stage of the pre-implantation mouse embryo, we determined the sequence of full-length cDNA and defined properties of the genomic organization of the mouse eIF-1A gene. Northern blot analysis distinguishes three transcripts in mouse liver of 2.8, 2.2 and 1.9 kb in size. The three transcripts arise from initiation at two putative promoters separated by 627 bp. Initiation from the putative distal promoter yields both the 2.8 and 1.9 kb transcripts, in which the 1.9 kb transcript is generated by alternative splicing of 840 bp of intervening RNA. The putative distal promoter, which lacks both a TATA box and CCAAT box control elements but contains several GC-rich clusters, initiates transcription at two start sites that are separated by 30 bp. Thus, four transcripts are generated from the distal promoter. The putative proximal promoter that directs transcription of a single 2.2 kb mRNA is preceded by a TATA box element that binds TBP. Each of the promoters is used by the pre-implantation mouse embryo, since we have been able to amplify selectively each of the five individual eIF-1A transcripts initiated from each promoter and start site in the 2-cell mouse embryo. PMID- 9753745 TI - Prediction of locally optimal splice sites in plant pre-mRNA with applications to gene identification in Arabidopsis thaliana genomic DNA. AB - Prediction of splice site selection and efficiency from sequence inspection is of fundamental interest (testing the current knowledge of requisite sequence features) and practical importance (genome annotation, design of mutant or transgenic organisms). In plants, the dominant variables affecting splice site selection and efficiency include the degree of matching to the extended splice site consensus and the local gradient of U- and G+C-composition (introns being U rich and exons G+C-rich). We present a novel method for splice site prediction, which was particularly trained for maize and Arabidopsis thaliana. The method extends our previous algorithm based on logitlinear models by considering three variables simultaneously: intrinsic splice site strength, local optimality and fit with respect to the overall splice pattern prediction. We show that the method considerably improves prediction specificity without compromising the high degree of sensitivity required in gene prediction algorithms. Applications to gene identification are illustrated for Arabidopsis and suggest that successful methods must combine scoring for splice sites, coding potential and similarity with potential homologs in non-trivial ways. A WWW version of the SplicePredictor program is available at http:/gnomic.stanford.edu/volker/SplicePredi ctor.html/ PMID- 9753746 TI - Inhibition of RNA polymerase III transcription by a ribosome-associated kinase activity. AB - Ribosomes prepared from somatic tissue of Xenopus laevis inhibit transcription by RNA polymerase III. This observation parallels an earlier report that a high speed fraction from activated egg extract, which is enrichedin ribosomes, inhibits RNA polymerase III activityand destabilizes putative transcription complexes assembled on oocyte 5S rRNA genes. Transcription of somatic- and oocyte type 5S rRNA genes and a tRNA gene are all repressed in the present experiments. We find that 5S rRNA genes incubated in S150 extract prepared from immature oocytes exhibit an extensive DNase I protection pattern that is nearly identical to that of the ternary complex of TFIIIA and TFIIIC bound to a somatic 5S rRNA gene. The complexes formed in this extract are stable at concentrations of ribosomes that completely repress transcription, indicating that formation of the TFIII(A+C) complex is not the target of inhibition. Ribosomes taken through a high salt treatment no longer repress transcription of class III genes, establishing that the inhibition is due to an associated factor and not the particle itself. The inhibitory activity released from ribosomes is inactivated by treatment with proteinase K, but not micrococcal nuclease. Preincubation of ribosomes with a general protein kinase inhibitor, 6-dimethylaminopurine, eliminates repression of transcription. Western blot analysis demonstrates that p34(cdc2), which is known to mediate repression of transcription by RNA polymerase III, is present in these preparations of ribosomes and can be released from the particles upon extraction with high salt. These results establish that a kinase activity, possibly p34(cdc2), is the actual agent responsible for the observed inhibition of transcription by ribosomes. PMID- 9753747 TI - Stabilization of slow troponin C polypeptide compensates for its reduced synthesis in antisense oligodeoxynucleotide-treated cells. AB - The expression of genes for contractile proteins during myogenesis is coordinately regulated. Uncoupling the expression of the slow/cardiac troponin C (sTnC) gene from this process with an antisense phosphorothioate oligodeoxynucleotide (ODN) was used to examine the presence of any post transcriptional mechanisms for regulating muscle protein synthesis. Approximately 70 and 50% decreases in sTnC polypeptide synthesis and mRNA levels, respectively, were achieved after 4 days antisense treatment. This decrease in sTnC polypeptide synthesis was not reflected in a similar decline in the steady-state level of this polypeptide. Extension of the ODN treatment to 7 days was required to produce a substantial decrease in the steady-state level of sTnC polypeptide. Our investigation suggests that during the 4-day treatment, the affected cells stabilized the sTnC polypeptide level by increasing its half-life. However, the stabilizing effect appears to be overridden during prolonged (7 days) antisense ODN treatment. Measurement of the polypeptide synthesis and mRNA levels of several contractile proteins showed no evidence of cross-regulation among the genes to coordinately regulate their expression levels. PMID- 9753748 TI - Interaction of myocyte enhancer factor 2 (MEF2) with a mitogen-activated protein kinase, ERK5/BMK1. AB - Myocyte enhancer factor 2 (MEF2) has been implicated in the complex hierarchical regulation of muscle-specific gene expression and differentiation. While the MyoD family members are able to initiate the skeletal muscle differentiation program, whether MEF2 is sufficient in directing skeletal muscle differentiation is still controversial. Furthermore, how MEF2 transactivates its target genes is not fully understood. It has been suggested that the interactions of MEF2 with other factors modify its transcriptional activity. Therefore, the identification of MEF2-interacting factors may be important in understanding the mechanism by which MEF2 activates its target genes. In this study, a mitogen-activated protein kinase (MAP kinase), ERK5/BMK1 was found to interact with MEF2 in a yeast two hybrid screen. The interaction was confirmed by a glutathione S -transferase-pull down assay and a co-immunoprecipitation study indicating that endogenous ERK5 and MEF2 interact with each other in vivo . The interacting domain of MEF2 was mapped to the N-terminus which contains the highly conserved MADS and MEF2 domains. Functionally, ERK5/BMK1 was able to phosphorylate MEF2 in vitro . Furthermore, when cotransfected with ERK5/BMK1, the transactivation capacity of MEF2 was enhanced. These results suggest that the functions of MEF2 could be regulated through ERK5/BMK1. PMID- 9753749 TI - Silent mutations in the Escherichia coli ompA leader peptide region strongly affect transcription and translation in vivo. AB - In order to test the effect of silent mutations on the regulation of gene expression, we monitored several steps of transcription and translation of the ompA gene in vivo , in which some or all codons between codons 6 and 14, frequently used in Escherichia coli , had been exchanged for infrequent synonymous codons. Northern blot analysis revealed an up to 4-fold reduction in the half-life of the mutated messengers and a >10-fold reduction in their steady state amounts. Western blot analysis showed a 10-fold reduction in the amount of OmpA protein. Use of a system expressing a Rho-specific anti-terminator allowed us to detect a strong transcription polarity effect in the silent mutants. These results demonstrate that silent mutations can severely inhibit several steps of gene expression in E. coli and that code degeneracy is efficiently exploited in this species for setting signals for gene control and regulation. PMID- 9753750 TI - Use of gap repair in fission yeast to obtain novel alleles of specific genes. AB - We have adapted a method for making libraries of mutations in any specific gene for use in the fission yeast Schizosaccharomyces pombe . This elegant and simple method consists of PCR amplification of the gene of interest, followed by co transformation of fission yeast with the PCR fragment and a linearized plasmid vector prepared such that the ends of the vector share DNA sequence with the ends of the PCR fragment. Homologous recombination between the vector and the PCR fragment occurs at a high frequency and results in a collection of yeast transformants, most harboring a mutated allele of the original gene within the vector of choice. This library can then be screened or selected for phenotypes of interest. PMID- 9753751 TI - A convenient method of aligning large DNA molecules on bare mica surfaces for atomic force microscopy. AB - Large DNA molecules remain difficult to be imaged by atomic force microscopy (AFM) because of the tendency of aggregation. A method is described to align long DNA fibers in a single direction on unmodified mica to facilitate AFM studies. The clear background, minimal overstretching, high reproducibility and convenience of this aligning procedure make it useful for physical mapping of genome regions and the studies of DNA-protein complexes. PMID- 9753752 TI - Quantitative hybridization to genomic DNA fractionated by pulsed-field gel electrophoresis. AB - Hybridization to genomic DNA fractionated by CHEF electrophoresis can vary >100 fold if the DNA is acid depurinated prior to Southern blotting. The level of hybridization is high or low depending on whether the molecule being analyzed migrates at a size coincident with or different from the size of the majority of genomic DNA in the sample, respectively. Techniques that avoid acid depurination including in-gel hybridizations and UV irradiation of DNA prior to blotting provide more accurate quantitative results. CHEF analysis of DNA molecules containing repetitive satellite sequences is particularly prone to this effect. PMID- 9753753 TI - Symptomatic scapulothoracic crepitus and bursitis. AB - Scapulothoracic crepitus and scapulothoracic bursitis are related painful disorders of the scapulothoracic articulation. Scapulothoracic crepitus is the production of a grinding or snapping noise with scapulothoracic motion, which may be accompanied by pain. Scapulothoracic bursitis manifests as pain and swelling of the bursae of the scapulothoracic articulation. Scapulothoracic bursitis is always seen in patients with symptomatic scapulothoracic crepitus, but may exist as an isolated entity. Symptomatic scapulothoracic crepitus may be due to pathologic changes in the bone or soft tissue between the scapula and the chest wall or may be due to changes in congruence of the scapulothoracic articulation, as seen in scoliosis and thoracic kyphosis. Treatment of patients with symptomatic scapulothoracic crepitus begins with nonoperative methods, including postural and scapular strengthening exercises and the application of local modalities. When soft-tissue lesions are the cause of scapulothoracic crepitus, conservative treatment is highly effective. When symptomatic scapulothoracic crepitus is due to osseous lesions, or when conservative treatment has failed, surgical options are available. Partial scapulectomies have produced satisfactory outcomes in selected patients. Recently, open and arthroscopic scapulothoracic bursectomies have been performed with some success and are being used more frequently. PMID- 9753754 TI - Tarsal coalition and painful flatfoot. AB - The prevalence of tarsal coalition is probably 1% or less. The two sites most commonly affected are the calcaneonavicular joint and the middle facet of the talocalcaneal joint. Diagnosis should be suspected in the preteen or teenage patient with insidious or sudden onset of pain in the midfoot to hindfoot associated with a lack of motion in the subtalar joint. Initial treatment with immobilization or an orthosis may relieve symptoms, but most patients will have persistent symptoms that warrant surgical correction. Long-term results indicate that excision of the coalition is moderately successful in relieving symptoms in the calcaneonavicular bar. Long-term success with excision of subtalar bars is less clear, although early relief of symptoms is usually possible. PMID- 9753755 TI - Compressive ulnar neuropathies at the elbow: I. Etiology and diagnosis. AB - Ulnar nerve compression at the elbow can occur at any of five sites that begin proximally at the arcade of Struthers and end distally where the nerve exits the flexor carpi ulnaris muscle in the forearm. Compression occurs most commonly at two sites-the epicondylar groove and the point where the nerve passes between the two heads of the flexor carpi ulnaris muscle (i.e., the true cubital tunnel). The differential diagnosis of ulnar neuropathies at the elbow includes lesions that cause additional proximal or distal nerve compression and systemic metabolic disorders. A complete history and a thorough physical examination are essential first steps in establishing a correct diagnosis. Electrodiagnostic studies may be useful, especially when the site of compression cannot be determined by physical examination, when compression may be at multiple levels, and when there are systemic and metabolic problems. PMID- 9753756 TI - Compressive ulnar neuropathies at the elbow: II. treatment. AB - Initial treatment of most compressive neuropathies at the elbow is nonoperative, consisting of rest, avoidance of elbow flexion, and, when necessary, temporary immobilization of the elbow and wrist. If symptoms persist, particularly when accompanied by muscle weakness, surgery is usually indicated. Operative procedures include decompression without transposition of the nerve (in situ or by means of medial epicondylectomy) and decompression with transposition of the nerve carried out in a subcutaneous, intramuscular, or submuscular fashion. The indications, advantages, disadvantages, and surgical technique of each operative procedure are discussed. PMID- 9753757 TI - Extremity fractures in the patient with a traumatic brain injury. AB - Extremity fractures are common in patients with traumatic brain injuries (TBIs). These injuries are often inadequately treated and occasionally are completely missed due to the unique problems inherent to the TBI patient. However, appropriate evaluation of the TBI patient allows prompt diagnosis and optimal treatment of extremity fractures. The increased survival rate of these patients has resulted in a greater emphasis on minimizing dysfunction and disability, especially that due to concomitant orthopaedic trauma. Advances in anesthestic technique permit earlier operative fixation of extremity fractures. Most injuries, particularly those in the lower extremity, require operative stabilization to allow early mobilization and rehabilitation. Upper extremity fractures are often associated with peripheral nerve injuries. Heterotopic ossification is common, especially about the elbow and hip. Contrary to prevalent belief, fracture healing is not necessarily accelerated in the TBI patient; hypertrophic callus, myositis ossificans, and heterotopic ossification occur frequently and are often misperceived as accelerated healing. PMID- 9753758 TI - Total wrist arthroplasty. AB - Although arthroplasty is a well-established procedure for many joints, its use in the wrist is less common, and the indications are less well defined. The standard procedure for the painful arthritic wrist remains radiocarpal arthrodesis. However, as technology and surgical procedures improve, wrist arthroplasty is being used more frequently. The authors provide a brief history of total wrist arthroplasty and review the arthroplasties most commonly used in the United States. Results with total wrist implants, the complications related to arthroplasty, technical aspects of the procedure, and salvage options are also discussed. PMID- 9753759 TI - Achilles tendon injuries. AB - As the number of persons who participate in athletic activity into their later years has increased, so has the incidence of overuse injuries to the Achilles tendon. The etiology of these problems is multifactorial and includes biomechanical factors and training errors. Use of a histopathologic scheme for classification of these injuries facilitates a logical approach to treatment. Conservative care is a mainstay of treatment for inflammatory conditions. Satisfactory outcomes may be obtained with either nonoperative or operative treatment of acute ruptures, although surgically treated patients appear to recover better functional capacity. Treatment of neglected injuries to the Achilles tendon continues to be a challenging problem. PMID- 9753760 TI - Determination and estimation of water solubilities and octanol/water partition coefficients for derivates of benzanilides. AB - A shake-flask method was used to determine the water solubilities (Sw) and the octanol/water partition coefficients(Kow) of 14 derivates of benzanilides. The results showed that the octanol/water partition coefficients were correlated (r= 0.93) with Sw. The linear solvation energy relationship(LSER) and molecular connectivity methods have been used to establish correlation equation successfully. The LSER method was used to predict these partitioning properties more accurately. The estimated values were well fitted with observed ones. PMID- 9753761 TI - Dioxin-like PCBs in the environment-human exposure and the significance of sources. AB - Dioxin-like PCBs represent an important component of the Sigma-TEQ in many environmental media. Specifically, in animal produce and in fish PCBs dominate the Sigma-TEQ ingested by humans. This in turn leads to high background body burdens in humans with PCB-TEQ greater than that associated with PCDD/Fs. High fish consumers are apparently subject to elevated TEQ exposure from dioxin-like PCBs. This has important implications for exposure assessment studies which have previously only been concerned with PCDDs and PCDFs. Unlike PCDD/Fs, dioxin-like PCBs are not controlled within the food chain. Sources and pathways of exposure are poorly defined. Aroclor formulations and their subsequent usage are considered to be the most important sources in terms of human exposure to some TEF-rated congeners, notably PCB-118, PCB-156 and part of PCB-126. Emissions from combustion sources contribute additional PCB-126. More research is needed to place these compounds in an integrated risk evaluation framework. PMID- 9753762 TI - Change and behavior of residual mercury in paddy soils and rice of Japan. AB - Paddy soils collected near Agano River, Niigata Prefecture in Japan. Agano River is located on an attack area of Niigata-Minamata disease. Soil samples were collected from 122 sites in 1989 and 120 sites in 1997. As a result, the total mercury concentrations were ranged from 0.019 to 0.62ppm based on dry weight with a mean 0.155ppm in 1989, and from 0.015 to 0.34 with a mean 0.146ppm in 1997. The decrease of residual mercury concentrations in paddy soils was only 0.009ppm in the interval of eight years. However, the mercury concentrations in paddy soils were about 3 times as large as that of uncultivated soils in its surroundings. It is suggested that the soils in paddy fields still contain the mercury residues to be influenced by some agricultural fungicide. Mercury concentrations in rice were natural background level. A comprehensive evaluation shows that the concentrations in boil rice with an average content of under 0.001ppm total mercury pose no health risk to Japanese people consuming rice of 200g/day. PMID- 9753763 TI - Relationship between organochlorine contaminants and mixed function oxidase activity in skin biopsy specimens of Mediterranean fin whales (Balaenoptera physalus). AB - The relationship between organochlorine contaminants (PCBs and DDTs) and mixed function oxidase, benzo(a)pyrene monooxigenase activity (BPMO), was investigated in skin biopsy specimens from fin whales (Balaenoptera physalus) of the Mediterranean Sea. Skin biopsy material, sampled by a non invasive technique, is suitable for a wide range of chemical and biomarker analysis. In this study PCBs and DDTs were evaluated in subcutaneous blubber and MFO activity in epidermis. An interesting correlation was found in male specimens between the two variables. PMID- 9753764 TI - Increased transcription of CYP6D1 causes cytochrome P450-mediated insecticide resistance in house fly. AB - Insecticide resistance is a major problem that continues to plague efforts to control pests of animals and crops. An important mechanism by which insects become resistant to insecticides is via increased detoxification mediated by the cytochrome P450 microsomal monooxygenases (monooxygenases). One of the fundamental gaps in our knowledge about this resistance mechanism is an understanding of how insects express high levels of the specific cytochrome P450(s) responsible for resistance. One such P450, CYP6D1, causes resistance to pyrethroid in the house fly and is expressed at 9-fold higher levels (mRNA and protein) in the Learn Pyrethroid Resistant (LPR) strain (compared to susceptible strains). The relative stability of CYP6D1 mRNA in resistant and susceptible strains was measured following inhibition of transcription with actinomycin D. The same time course of decrease in CYP6D1 mRNA abundance was detected in both strains indicating that the high level of expression of CYP6D1 in LPR is not due to increased stability of the mRNA. The comparative rates of transcription of CYP6D1 were measured using an in vitro run-on transcription assay. The relative amount of CYP6D1 transcript produced in this assay was 10-fold greater in the LPR strain compared to the susceptible strain. This demonstrates that increased transcription of CYP6D1 is an underlying cause of monooxygenase-mediated insecticide resistance. The increased rate of transcription of CYP6D1 in the resistant strain (LPR) is controlled by two factors: one on autosome 1 and another on autosome 2. PMID- 9753765 TI - Nucleotide sequence of vitellogenin mRNA in the bean bug, Riptortus clavatus: analysis of processing in the fat body and ovary. AB - The bean bug, Riptortus clavatus, has immunologically distinct yolk proteins, vitellin (Vn)-1 and-2 and their precursors, vitellogenin (Vg)-1 and-2. We have cloned the full nucleotide sequence of Vg-1 cDNA. The deduced amino acid sequence has some similarities to other insect Vgs. It contains two polyserine regions, which are characteristic of other Vgs. Vg-1 mRNA appeared after treatment with the juvenile hormone analogue, methoprene, implying transcriptional regulation. We found four enzymatic cleavage sites in the Vg molecule. Two of them match the consensus for dibasic processing endoprotease, which is also conserved in processing sites for other insect Vgs. We showed that the processing at each site was incomplete, and this resulted in production of more than the five polypeptides which would be expected from four processing sites in the molecule. The physiological significance of multiple polypeptides in insect Vgs is still unclear. PMID- 9753766 TI - The cysteine protease activity of Colorado potato beetle (Leptinotarsa decemlineata Say) guts, which is insensitive to potato protease inhibitors, is inhibited by thyroglobulin type-1 domain inhibitors. AB - High levels of protease inhibitors are induced in potato leaves by wounding. These inhibitors, when ingested by Colorado potato beetle (Leptinotarsa decemlineata Say) larvae, induce expression of specific proteolytic activities in the gut. Induced protease activities cannot be inhibited by potato inhibitors and thus enable the insects to overcome this defence mechanism of potato plants. The induced aminopeptidase and endoproteolytic activities both have the characteristics of cysteine proteases. Twenty-one protein inhibitors of different structural types have been examined for their ability to inhibit these activities in vitro. Members of the cystatin superfamily were found to be poor inhibitors of the induced endoproteolytic activities, except for the third domain of human kininogen, which was a fairly strong inhibitor (75% inhibition). The strongest inhibition (85%) of induced endoproteolytic activity was obtained using structurally different thyroglobulin type-1 domain-like inhibitors--equistatin and MHC class II-associated p41 invariant fragment. Experiments performed using three synthetic substrates for endoproteases gave similar results and indicate the existence of at least different endoproteolytic enzymes resistant to potato inhibitors. The induced aminopeptidase activity can be inhibited only by stefin family of inhibitors in cystatin superfamily. In in vivo experiments, Colorado potato beetle larvae fed on equistatin-coated potato leaves were strongly retarded in their growth and almost 50% died after 4 days. This demonstrated the potential of equistatin to protect crops from insect attack. PMID- 9753767 TI - Isolation and characterization of two cytochrome P450 cDNA clones for CYP6B6 and CYP6B7 from Helicoverpa armigera (Hubner): possible involvement of CYP6B7 in pyrethroid resistance. AB - Two new cDNA clones specific for members of the CYP6B gene family, CYP6B6 and CYP6B7 have been isolated from Helicoverpa armigera. The sequences correspond to mRNAs of an estimated 1962 and 2411 nucleotides in length respectively excluding the poly A tails. The two mRNAs have open reading frames encoding proteins of 504 amino acid residues with molecular weights of 57,564 and 58,181 Daltons. Both putative proteins contain the conserved cysteine and surrounding regions characteristics of all cytochrome P450s. The encoded protein sequences show 84 88% protein sequence identity between them and with the previously published cytochrome P450 sequence of H. armigera. CYP6B2. The sequences of cDNA clones of CYP6B6 and CYP6B2 show a very high degree of identity within the first 340 nucleotides which may be the result of a gene conversion event. Two major bands are visible after northern analysis of larval RNA using cDNA clones for CYP6B6, CYP6B7, or the previously published CYP6B2 as probes, due to strong cross hybridization. Analysis with specific oligonucleotide probes and 3' non-coding regions indicated that the cDNAs for CYP6B6 and CYP6B7 correspond to the smaller and large mRNA bands respectively. The previously identified sequence of CYP6B2, contrary to the previous suggestion, corresponds to a rare mRNA of similar size to that for CYP6B6. The mRNA for CYP6B7 was found to be induced by treatment with the monoterpene, alpha-pinene, and to be over-expressed in some individuals of pyrethroid resistant population of H. armigera. We suggest that CYP6B7 is the form responsible for pyrethroid metabolism in H. armigera. PMID- 9753768 TI - Acetylcholinesterase cDNA of the cattle tick, Boophilus microplus: characterisation and role in organophosphate resistance. AB - Acetylcholinesterase is the target of organophosphate and carbamate pesticides. Organophosphate resistance is widespread in the cattle tick, Boophilus microplus, in Australia. We have isolated a cDNA of acetylcholinesterase from B. microplus and show that it would encode a protein 62 kDa in size. The predicted amino acid sequence contains all the residues characteristic of an acetylcholinesterase. Alternative splicing of the transcript was detected at both the 5' and 3' ends. Alternative splicing at the 5' end would result in two proteins differing by six amino acids. This is the first report of alternative splicing of the N-terminal coding region in a cholinesterase. No point mutations were detected in the acetylcholinesterase gene from organophosphate resistant strains of B. microplus. Alternative explanations for resistance to organophosphates in B. microplus are discussed. PMID- 9753769 TI - The pheromone biosynthesis activating neuropeptide (PBAN) of the black cutworm moth, Agrotis ipsilon: immunohistochemistry, molecular characterization and bioassay of its peptide sequence. AB - PBAN-like immunoreactivity has been detected in the suboesophageal ganglion and the brain (Br-SOG) of larvae and adult males and females of Agrotis ipsilon, using an antiserum against Helicoverpa zea PBAN (Hez-PBAN). The amino acid sequence of A. ipsilon PBAN (Agi-PBAN) was deduced from the cDNA sequence, using both Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) and 5' Rapid Amplification of cDNA Ends (RACE). The primers were degenerate sets of oligonucleotides derived from known amino acid sequences of the PBAN precursor. The final cloned fragment contained the complete DNA sequence coding for the putative Agi-PBAN. Based on a comparison with known PBAN processing from the polypeptide precursor, we propose that Agi-PBAN is a 33-amino acid peptide. Agi PBAN exhibits high sequence homology with Hez-PBAN (88%), Lymantria dispar PBAN (Lyd-PBAN, 88%) and Bombyx mori PBAN (Bom-PBAN, 73%). Agi-PBAN shares the C terminal hexapeptide sequence (Tyr-Phe-Ser-Pro-Arg-LeuNH2) with all identified PBANs but has only one methionine residue instead of two in Hez-PBAN and Lyd PBAN, and three in Bom-PBAN. Based on predicted a.a. sequence, Agi-PBAN, with Leu NH2 as C-terminal motif, has been synthesized and assayed for its ability to promote pheromone production in decapitated females of A. ipsilon. Synthetic Agi PBAN induced pheromone production in decapitated females as evaluated by the male responsiveness to the pheromonal blend in a wind tunnel. PMID- 9753770 TI - Molecular characterization of the Notch homologue from the Australian sheep blowfly, Lucilia cuprina. AB - The Drosophila melanogaster Notch gene product as a receptor of intercellular signals and is central to cell fate specification. The Scalloped wings (Scl) gene is the homologue of Notch in the Australian sheep blowfly, Lucilia cuprina. An allele of Scl is thought to be involved in the modification of Darwinian fitness and bristle asymmetry in flies resistant to organophosphorous chemicals (OPs). As a first step towards the testing of this hypothesis we cloned and sequenced Scl. A full-length cDNA segment representing the mRNA of Scl is 8503 bp and encodes a protein of 2653 amino acids, which shares 73.6% identity with Notch. All functional motifs including EGF-like repeats, LNR repeats, cdc 10/ankyrin repeats, opa and PEST elements are present in the same order as in Notch and the sequence identities peak in these motifs. With respect to genomic structure, intron/exon boundaries are conserved but, in most cases, the Scl introns are larger. Sequence analysis of the upstream genomic region reveals that the gene has a TATA-less promoter. Consistent with a central role in embryogenesis and imaginal development, high levels of Scl expression were detected in the early embryonic and pupal stages. PMID- 9753771 TI - Similarities to a LINE element shared by Anopheline and Culicine mosquitos map to the distal end of dihydrofolate reductase amplicons in Aedes albopictus mosquito cells. AB - To extend our understanding of amplicon structure in methotrexate-resistant Mtx 5011-256 Aedes albopictus mosquito cells, we examined a series of cosmids containing genomic DNA corresponding to the unique 3'-end of the Type 1 dihydrofolate reductase amplicon. Cosmid pWED118 contained five EcoRI fragments ranging from 2 to 5 kb (A, B, C, F, G) that hybridized to cDNA from methotrexate resistant cells. Of these, fragments B and F hybridized weakly to first-strand cDNA from sensitive cells and shared considerable sequence identity. Fragment G occurred twice in the map of pWED118; one copy mapped within a 10 kb BssHII core fragment from the Type I amplicon and a second copy mapped downstream in the 48 kb BssHII core fragment. Hybridization signals among fragments contained in overlapping cosmids suggested that a branch point defining two or more subtypes of the Type I amplicon occurs within or near the 10 kb BssHII genomic DNA fragment. A 1.8 kb sequence common to fragments B and F included an approximately 0.4 kb region that shared sequence similarities with a LINE element from Aedes aegypti and with a repeated sequence from Anopheles gambiae. In addition, these elements shared amino acid similarity to a reverse transcriptase from the nematode, Caenorhabditis elegans. Shared sequence between Aedes and Anopheles elements supports the hypothesis that an ancestral LINE-like element was active in mosquito genomes prior to the divergence of the subfamilies Culicinae and Anophelinae. The presence of homologies to LINE-like elements near a branch point in the dihydrofolate reductase amplicon is consistent with a possible role of repeated sequences in amplicon shortening. PMID- 9753772 TI - Specific accumulation of GFP in a non-acidic vacuolar compartment via a C terminal propeptide-mediated sorting pathway. AB - The green fluorescent protein (GFP) from Aequorea victoria can be detected in living plant cells after transient transformation of protoplasts. Expression of the GFP can be used to monitor protein trafficking in a mixed cell population and also to study the different function and importance of organelles in different cell types. We developed a vacuolar form of GFP that was obtained by replacing the C-terminal endoplasmic reticulum (ER)-retention motif of mGFP5-ER by the vacuolar targeting peptide of tobacco chitinase A. The vacuolar GFP was transported and accumulated in the vacuole as expected. However, we found two patterns of GFP accumulation after prolonged incubation (18-24 h) depending on the cell type. Most chloroplast-rich protoplasts had a fluorescent large central vacuole. In contrast, most chloroplast-poor protoplasts accumulated the GFP in one smaller vacuole but not in the large central vacuole, which was visible under a light microscope in the same cell. This differential accumulation reflected the existence of two different vacuolar compartments as described recently by immunolocalization of several vacuolar markers. We were able to characterize the vacuolar compartment to which GFP is specifically targeted as non-acidic, since it did not accumulate neutral red while acidic vacuoles did not accumulate GFP. PMID- 9753773 TI - Expression of snowdrop lectin (GNA) in transgenic rice plants confers resistance to rice brown planthopper. AB - Snowdrop lectin (Galanthus nivalis agglutinin; GNA) has been shown previously to be toxic towards rice brown planthopper (Nilaparvata lugens; BPH) when administered in artificial diet. BPH feeds by phloem abstraction, and causes 'hopper burn', as well as being an important virus vector. To evaluate the potential of the gna gene to confer resistance towards BPH, transgenic rice (Oryza sativa L.) plants were produced, containing the gna gene in constructs where its expression was driven by a phloem-specific promoter (from the rice sucrose synthase RSs1 gene) and by a constitutive promoter (from the maize ubiquitin ubi1 gene). PCR and Southern analyses on DNA from these plants confirmed their transgenic status, and that the transgenes were transmitted to progeny after self-fertilization. Western blot analyses revealed expression of GNA at levels of up to 2.0% of total protein in some of the transgenic plants. GNA expression driven by the RSs1 promoter was tissue-specific, as shown by immunohistochemical localization of the protein in the non-lignified vascular tissue of transgenic plants. Insect bioassays and feeding studies showed that GNA expressed in the transgenic rice plants decreased survival and overall fecundity (production of offspring) of the insects, retarded insect development, and had a deterrent effect on BPH feeding. gna is the first transgene to exhibit insecticidal activity towards sap-sucking insects in an important cereal crop plant. PMID- 9753774 TI - Cloning of the fructan biosynthesis pathway of Jerusalem artichoke. AB - To study the regulation of fructan synthesis in plants, we isolated two full-size cDNA clones encoding the two enzymes responsible for fructan biosynthesis in Jerusalem artichoke (Helianthus tuberosus): 1-sucrose:sucrose fructosyl transferase (1-SST) and 1-fructan:fructan fructosyl transferase (1-FFT). Both enzymes have recently been purified to homogeneity from Jerusalem artichoke tubers (Koops and Jonker (1994) J.Exp.Bot.45, 1623-1631; Koops and Jonker (1996) Plant Physiol. 110, 1167-1175) and their amino acid sequences have been partially determined. Using RT-PCR and primers based on these sequences, specific fragments of the genes were amplified from tubers of Jerusalem artichoke. These fragments were used as probes to isolate the cDNAs encoding 1-SST and 1-FFT from a tuber specific lambdal ZAP library. The deduced amino acid sequences of both cDNAs perfectly matched the sequences of the corresponding purified proteins. At the amino acid level, the cDNA sequences showed 61% homology to each other and 59% homology to tomato vacuolar invertase. Based on characteristics of the deduced amino acid sequence, the first 150 bp of both genes encode a putative vacuolar targeting signal. Southern blot hybridization revealed that both 1-SST and 1-FFT are likely to be encoded by single-copy genes. Expression studies based on RNA blot analysis showed organ-specific and developmental expression of both genes in growing tubers. Lower expression was detected in flowers and in stem. In other organs, including leaf, roots and dormant tubers, no expression could be detected. In tubers, the spatial and developmental expression correlates with the accumulation of fructans. Using the 1-sst and 1-fft cDNAs, chimeric genes were constructed driven by the CaMV 35S promoter. Analysis of transgenic petunia plants carrying these constructs showed that both cDNAs encode functional fructosyltransferase enzymes. Plants transformed with the 35S-1-sst construct accumulated the oligofructans 1-kestose (GF2), 1,1-nystose (GF3) and 1,1,1 fructosylnystose (GF4). Plants transformed with the 35S-1-fft construct did not accumulate fructans, probably because of the absence of suitable substrates for 1 FFT, i.e. fructans with a degree of polymerization > or = 3 (GF2, GF3, etc.). Nevertheless, protein extracts from these transgenic plants were able to convert GF3, when added as a substrate into fructans with a higher degree of polymerization. Progeny of crosses between a 35S-1-sst-containing plant and a 35S 1-fft-containing plant, showed accumulation of high-molecular-weight fructans in old, senescent leaves. Based on the comparison of the predicted amino acid sequences of 1-sst and 1-fft with those of other plant fructosyl transferase genes, we postulate that both plant fructan genes have evolved from plant invertase genes. PMID- 9753775 TI - ICK1, a cyclin-dependent protein kinase inhibitor from Arabidopsis thaliana interacts with both Cdc2a and CycD3, and its expression is induced by abscisic acid. AB - Cyclin-dependent kinase (CDK) inhibitor genes encode low molecular weight proteins which have important functions in cell cycle regulation, development and perhaps also in tumorigenesis. The first plant CDK inhibitor gene ICK1 was recently identified from Arabidopsis thaliana. Although the C-terminal domain of ICK1 contained an important consensus sequence with the mammalian CDK inhibitor p27Kip1, the remainder of the deduced ICK1 sequence showed little similarity to any known CDK inhibitors. In vitro assays showed that recombinant ICK1 exhibited unique kinase inhibitory properties. In the present study we characterized ICK1 in terms of its gene structure, its interaction with both A. thaliana Cdc2a and CycD3, and its induction by the plant growth regulator, abscisic acid (ABA). ICK1 was expressed at a relatively low level in the tissues surveyed. However, ICK1 was induced by ABA, and along with ICK1 induction there was a decrease in Cdc2 like histone H1 kinase activity. These results suggest a molecular mechanism by which plant cell division might be inhibited by ABA. ICK1 clones were also identified from independent yeast two-hybrid screens using the CycD3 construct. The implication that ICK1 protein could interact with both Cdc2a and CycD3 was confirmed by in vitro binding assays. Furthermore, deletion analysis indicated that different regions of ICK1 are required for the interactions with Cdc2a and CycD3. These results provide a mechanistic basis for understanding the role of CDK inhibitors in cell cycle regulation in plant cells. PMID- 9753776 TI - Molecular characterization of a plant FKBP12 that does not mediate action of FK506 and rapamycin. AB - Immuonosuppressive drugs FK506 and rapamycin block a number of signal transduction pathways in eukaryotic systems. The 12 kDa FK506 binding protein (FKBP12) mediates the action of both FK506 and rapamycin against their functional targets. In this report, we cloned, sequenced and characterized a gene encoding FKBP12 in Vicia faba (VfFKBP12). While VfFKBP12 is highly homologous to animal and yeast FKBP12, it does not mediate the action of FK506 and rapamycin. There are unique features in plant FKBP12 sequences that cause the variation in their function. One lies in the domain that is critical for interaction with calcineurin (CaN), the mammalian and yeast target of FKBP12-FK506 complex. Protein-protein interaction assays revealed a low-affinity and unstable VfFKBP12 FK506-CaN ternary complex. In the genetic assay, VfFKBP12 did not restore the sensitivity of yeast FKBP12 mutant to rapamycin or FK506, supporting that plant FKBP12-ligand complexes are unable to block the function of the drug target. Also unique to plant FKBP12 proteins, a pair of cysteines is spatially adjacent to potentially form disulfide linkage. Treatment of VfFKBP12 with reductant dithiothreitol (DTT) abolished the formation of VfFKBP12-FK506-CaN ternary complex. Site-directed mutagenesis to substitute one of the cysteines, Cys26, with Ser produced a similar effect as DTT treatment. These results indicate that an intramolecular disulfide bond is a novel structural feature required for the low-affinity interaction between plant FKBP12 and CaN. In conclusion, plant FKBP12 proteins have evolved structural changes that modify their protein-protein interacting domains and cause loss of function against the drug targets. PMID- 9753777 TI - Polyunsaturated membranes are required for photosynthetic competence in a mutant of Arabidopsis. AB - High levels of polyunsaturation are characteristic of all the membranes of plant and animal cells. For example, the chloroplasts of leaf cells contain about 75 80% polyunsaturated fatty acids. For the extra-chloroplast membranes in leaf cells and the membranes of non-photosynthetic tissues, values of 60-65% are typical. We report here the production of Arabidopsis double mutants that contain negligible levels of polyunsaturated fatty acids. The mutants were not capable of autotrophic growth and produced extremely chlorotic cotyledons and leaves. However, on sucrose media, the double mutants were robust plants showing strong leaf and root development. These observations indicate that the vast majority of receptor-mediated and transport-related membrane functions required to sustain the organism and induce proper development are adequately supported in the absence of polyunsaturated lipids. By contrast, photosynthesis is one process that does require high levels of membrane polyunsaturation. PMID- 9753778 TI - Two different genes encode ferrochelatase in Arabidopsis: mapping, expression and subcellular targeting of the precursor proteins. AB - Ferrochelatase is the last enzyme of haem biosynthesis. We have isolated 27 independent ferrochelatase cDNAs from Arabidopsis thaliana by functional complementation of a yeast mutant. Twenty-two of these cDNAs were similar to a previously isolated clone, AF3, and although they varied in length at the 5' and 3' ends, their nucleotide sequences were identical, indicating that they were derived from the same gene (ferrochelatase-I). The remaining five cDNAs all encoded a separate ferrochelatase isoform (ferrochelatase-II), which was 69% identical at the amino acid level to ferrochelatase-I. Using RFLP analysis in recombinant inbred lines, the ferrochelatase-I gene was mapped to chromosome V and that for ferrochelatase-II to chromosome II. Northern analysis showed that both ferrochelatase genes are expressed in leaves, stems and flowers, and expression in the leaves is higher in the light than in the dark. However, in roots only ferrochelatase-I transcripts were detected. High levels of sucrose stimulated expression of ferrochelatase-I, but had no effect, or repressed slightly, the expression of the ferrochelatase-II isoform. Import experiments into isolated chloroplasts and mitochondria showed that the ferrochelatase-II gene encodes a precursor which is imported solely into the chloroplast, in contrast to ferrochelatase-I which is targeted to both organelles. The significance of these results for haem biosynthesis and the production of haemoproteins, both within the plant cell and in different plant tissues, is discussed. PMID- 9753779 TI - Developmental abnormalities associated with deoxyadenosine methylation in transgenic tobacco. AB - As in other higher eukaryotes, DNA methylation in plants is predominantly found at deoxycytosine residues, while deoxyadenosine residues are not methylated at significant levels. 6mdA methylation has been successfully introduced into yeast and Drosophila via expression of a heterologous methyltransferase, but similar attempts in tobacco had, up until now, proved unsuccessful despite the correct expression of a methyltransferase construct. It was unclear whether this result reflected the failure of heterologous methyltransferases to enter the nucleus, or whether 6mdA methylation, which has been shown to interfere with promoter activity, was toxic for plants. Here we show that 6mdA methylation can be successfully introduced into transgenic tobacco plants via expression of the bacterial dam enzyme. The efficiency of 6mdA methylation was directly proportional to expression levels of the dam construct, and methylation of all GATC sites was observed in a highly expressing line. Increasing expression levels of the enzyme in different plants correlated with increasingly abnormal phenotypes affecting leaf pigmentation, apical dominance, and leaf and floral structure. Whilst introduction of dam-specific methylation does not cause any developmental abnormalities in yeast or Drosophila, our data suggest that methylation of deoxyadenine residues in plants interferes with the expression of genes involved in leaf and floral development. PMID- 9753780 TI - Co- and/or post-translational modifications are critical for TCH4 XET activity. AB - TCH4 encodes a xyloglucan endotransglycosylase (XET) of Arabidopsis thaliana. XETs endolytically cleave and religate xyloglucan polymers; xyloglucan is one of the primary structural components of the plant cell wall. Therefore, XET function may affect cell shape and plant morphogenesis. To gain insight into the biochemical function of TCH4, we defined structural requirements for optimal XET activity. Recombinant baculoviruses were designed to produce distinct forms of TCH4. TCH4 protein engineered to be synthesized in the cytosol and thus lack normal co- and post-translational modifications is virtually inactive. TCH4 proteins, with and without a polyhistidine tag, that harbor an intact N-terminus are directed to the secretory pathway. Thus, as predicted, the N-terminal region of TCH4 functions as a signal peptide. TCH4 is shown to have at least one disulfide bond as monitored by a mobility shift in SDS-PAGE in the presence of dithiothreitol (DTT). This disulfide bond(s) is essential for full XET activity. TCH4 is glycosylated in vivo; glycosidases that remove N-linked glycosylation eliminated 98% of the XET activity. Thus, co- and/or post-translational modifications are critical for optimal TCH4 XET activity. Furthermore, using site specific mutagenesis, we demonstrated that the first glutamate residue of the conserved DEIDFEFL motif (E97) is essential for activity. A change to glutamine at this position resulted in an inactive protein; a change to aspartic acid caused protein mislocalization. These data support the hypothesis that, in analogy to Bacillus beta-glucanases, this region may be the active site of XET enzymes. PMID- 9753781 TI - A gene encoding phosphatidylinositol-4-phosphate 5-kinase is induced by water stress and abscisic acid in Arabidopsis thaliana. AB - Phosphatidylinositol-4-phosphate 5-kinase (PIP5K) phosphorylates phosphatidylinositol-4-phosphate to produce phosphatidylinositol-4,5-bisphosphate as a precursor of two second messengers, inositol-1,4,5-triphosphate and diacylglycerol, and as a regulator of many cellular proteins involved in signal transduction and cytoskeletal organization. Despite PIP5K playing such an essential role in a number of physiological processes, much still remains to be made clear about its association with plants. Searching the Arabidopsis expression sequence tag database against already known yeast and mammalian PIP5K cDNAs, we identified two clones which partly encode the same Arabidopsis PIP5K and isolated a corresponding full-length cDNA encoding a protein that we designated AtPIP5K1. Recombinant AtPIP5K1 expressed in Escherichia coli possessed a PIP5K activity in vitro. Due to some structural and biochemical differences, AtPIP5K1 was not categorized as either a type I or type II PIP5K. The expression of the AtPIP5K1 mRNA was induced rapidly by treating Arabidopsis plants with drought, salt and abscisic acid, which suggests that AtPIP5K11 is involved in water-stress signal transduction. These data give evidence for a close link between phosphoinositide signaling cascades and water-stress responses in plants. PMID- 9753782 TI - Biases in three-dimensional structure-from-motion arise from noise in the early visual system. AB - The projected pattern of retinal-image motion supplies the human visual system with valuable information about properties of the three-dimensional environment. How well three-dimensional properties can be recovered depends both on the accuracy with which the early motion system estimates retinal motion, and on the way later processes interpret this retinal motion. Here we combine both early and late stages of the computational process to account for the hitherto puzzling phenomenon of systematic biases in three-dimensional shape perception. We present data showing how the perceived depth of a hinged plane ('an open book') can be systematically biased by the extent over which it rotates. We then present a Bayesian model that combines early measurement noise with geometric reconstruction of the three-dimensional scene. Although this model has no in built bias towards particular three-dimensional shapes, it accounts for the data well. Our analysis suggests that the biases stem largely from the geometric constraints imposed on what three-dimensional scenes are compatible with the (noisy) early motion measurements. Given these findings, we suggest that the visual system may act as an optimal estimator of three-dimensional structure-from motion. PMID- 9753783 TI - Auditory hair cell precursors immortalized from the mammalian inner ear. AB - Mammalian auditory hair cells are few in number, experimentally inaccessible, and do not proliferate postnatally or in vitro. Immortal cell lines with the potential to differentiate into auditory hair cells would substantially facilitate auditory research, drug development, and the isolation of critical molecules involved in hair cell biology. We have established two conditionally immortal cell lines that express at least five characteristic hair cell markers. These markers are the transcription factor Brn3.1, the alpha 9 subunit of the acetylcholine receptor, the stereociliary protein fimbrin and the myosins VI and VIIA. These hair cell precursors permit functional studies of cochlear genes and in the longer term they will provide the means to explore therapeutic methods of stimulating auditory hair cell regeneration. PMID- 9753784 TI - Multiple groups of orientation-selective visual mechanisms underlying rapid orientated-line detection. AB - Visual search for an edge or line element differing in orientation from a background of other edge or line elements can be performed rapidly and effortlessly. In this study, based on psychophysical measurements with ten human observers, threshold values of the angle between a target and background line elements were obtained as functions of background-element orientation, in brief masked displays. A repeated-loess analysis of the threshold functions suggested the existence of several groups of orientation-selective mechanisms contributing to rapid orientated-line detection; specifically, coarse, intermediate and fine mechanisms with preferred orientations spaced at angles of approximately 90 degrees, 35 degrees, and 10 degrees-25 degrees, respectively. The preferred orientations of coarse and some intermediate mechanisms coincided with the vertical or horizontal of the frontoparallel plane, but the preferred orientations of fine mechanisms varied randomly from observer to observer, possibly reflecting individual variations in neuronal sampling characteristics. PMID- 9753785 TI - Defaults in stereoscopic and kinetic depth perception. AB - This study presents three findings concerning the mechanisms of depth perception. First, the shape of the three-dimensional percept evoked by two-frame motion is defined solely by the rotation component around an axis in the frontoparallel plane; the visual system assigns a default value to this rotation component to arrive at a unique solution. Second, when the visual axes of two eyes are almost parallel, the visual system uses a default vergence value to reconstruct stereoscopic depth. Third, the default vergence and default rotation angles are highly correlated across subjects. This correlation implies that the two modalities share a common scaling default at an internal level. PMID- 9753786 TI - On the adaptive significance of stress-induced immunosuppression. AB - We approach the field of stress immunology from an ecological point of view and ask: why should a heavy physical workload, for example as a result of a high reproductive effort, compromise immune function? We argue that immunosuppression by neuroendocrine mechanisms, such as stress hormones, during heavy physical workload is adaptive, and consider two different ultimate explanations of such immunosuppression. First, several authors have suggested that the immune system is suppressed to reallocate resources to other metabolic demands. In our view, this hypothesis assumes that considerable amounts of energy or nutrients can be saved by suppressing the immune system; however, this assumption requires further investigation. Second, we suggest an alternative explanation based on the idea that the immune system is tightly regulated by neuroendocrine mechanisms to avoid hyperactivation and ensuing autoimmune responses. We hypothesize that the risk of autoimmune responses increases during heavy physical workload and that the immune system is suppressed to counteract this. PMID- 9753788 TI - Global matrilineal population structure in sperm whales as indicated by mitochondrial DNA sequences. AB - The genetic variability and population structure of worldwide populations of the sperm whale was investigated by sequence analysis of the first 5'L 330 base pairs in the mitochondrial DNA (mtDNA) control region. The study included a total of 231 individuals from three major oceanic regions, the North Atlantic, the North Pacific and the Southern Hemisphere. Fifteen segregating nucleotide sites defined 16 mtDNA haplotypes (lineages). The most common mtDNA types were present in more than one oceanic region, whereas ocean-specific types were rare. Analyses of heterogeneity of mtDNA type frequencies between oceans indicated moderate (GST = 0.03) but statistically significant (p = 0.0007) genetic differentiation on a global scale. In addition, strong genetic differentiation was found between potential social groups (GST = 0.03-0.6), indicating matrilineal relatedness within groups. The global nucleotide diversity was quite low (pi = 0.004) implying a recent common mtDNA ancestry (< 100,000) years ago) and a young global population structure. However, within this time period, female dispersal has apparently been limited enough to allow the development of global mtDNA differentiation. The results are consistent with those from observational studies and whaling data indicating stable social affiliations, some degree of area fidelity and latitudinal range limitations in groups of females and juveniles. PMID- 9753789 TI - A toxicity reduction evaluation for an oily waste treatment plant exhibiting episodic effluent toxicity. AB - A Toxicity Reduction Evaluation (TRE) was conducted on the oily wastewater treatment plant (Plant) at a Naval Fuel Depot. The Plant treats ship and ballast wastes, berm water from fuel storage areas and wastes generated in the fuel reclamation plant utilizing physical/chemical treatment processes. In the first period of the project (Period I), the TRE included chemical characterization of the plant wastewaters, monitoring the final effluent for acute toxicity and a thorough evaluation of each treatment process and Plant operating procedures. Toxicity Identification Evaluation (TIE) procedures were performed as part of the overall TRE to characterize and identify possible sources of toxicity. Several difficulties were encountered because the effluent was saline, test organisms were marine species and toxicity was sporadic and unpredictable. The treatability approach utilizing enhancements, improved housekeeping, and operational changes produced substantial reductions in the acute toxicity of the final effluent. In the second period (Period II), additional acute toxicity testing and chemical characterization were performed through the Plant to assess the long-term effects of major unit process improvements for the removal of toxicity. The TIE procedures were also modified for saline wastewaters to focus on suspected class of toxicants such as surfactants. The TRE was successful in reducing acute toxicity of the final effluent through process improvements and operational modifications. The results indicated that the cause of toxicity was most likely due to combination of pollutants (matrix effect) rather than a single pollutant. PMID- 9753790 TI - Determination of the uptake of [Pt(NH3)4](NO3)2 by grass cultivated on a sandy loam soil and by cucumber plants, grown hydroponically. AB - Two cultivation experiments were carried out in order to answer the question to what extent platinum can enter the food chain by accumulation in plants, when the platinum is present in a bio-available form: (i) cucumber plants (Cucumis sativus) were grown hydroponically in nutrient solutions containing [Pt(NH3)4](NO3)2 (from 0.5 to 50 micrograms Pt/l solution); and (ii) a water soluble platinum compound--[Pt(NH3)4](NO3)2--was added in increasing amounts to a sandy loam soil (from 0.5 to 50 mg Pt/kg soil) and rye grass (Lolium perenne) was grown on it. The roots on the one hand and the green plant fractions in the other hand of the cucumber plants and the rye grass were digested using a high-pressure asher. The platinum concentration was determined by means of a quadrupole-based (VG PQ I) or a double focusing sector field ICP-mass spectrometer (Finnigan MAT, Element), depending on the platinum concentration in the sample solution. The detection limit for platinum obtained with the VG PQ I was observed to be 6 ng/1, while with the 'Element' the detection limit could be improved to 0.5 ng/1 Pt. Accumulation factors were calculated as the ratio of the platinum concentration in the plant to that in the soil or the nutrient solution. The grass grown on spiked soil accumulated platinum only to a slight degree (accumulation factors between 0.008 and 0.032). The hydroponically grown cucumber plants, however, strongly accumulated it (accumulation factors of 11-42 in the shoot and 1700-2100 in the roots). There are three possible causes for the large differences in the accumulation factors: (i) Cucumber plants are dicotyledons; grass, however, is a monocotyledon. Other cultivation experiments already showed that dicotyledons accumulate metals to a higher extent than monocotyledons. (ii) In the grass cultivation experiment, the platinum compound was only added once to the sandy loam soil, namely 2 days before grass was cultivated on it. The nutrient solutions of the cucumber plants were changed twice a week. Consequently, the total amount of platinum that the plants were exposed to during the cultivation of the cucumber plants was higher than during the cultivation of the grass. (iii) Immobilization of the platinum compound in the soil most likely occurred. PMID- 9753791 TI - Temporal variation of PCDD/F concentrations in vegetation samples collected in the vicinity of a municipal waste incinerator (1996-1997). AB - In 1996, the concentrations of polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) were determined by HRGC/HRMS in 24 vegetation samples collected in the vicinity of a municipal solid waste incinerator (Tarragona, Catalonia, Spain). In the present study 24 vegetation samples were again taken at the same sampling points and analyzed for the levels of PCDD/Fs. The results were compared with those obtained in the previous survey. While in the 1996 study, PCDD/F levels ranged from 0.15 to 62.09 ng I-TEQ/kg (dry matter) (median value, 0.33 ng I-TEQ/kg; mean value, 4.11 ng I-TEQ/kg), the concentrations of PCDD/Fs in the 1997 survey ranged between 0.11 and 0.50 ng I-TEQ/kg (dry matter) (median value, 0.20 ng I-TEQ/kg; mean value, 0.23 ng I-TEQ/kg). The individual comparison between PCDD/F levels in the samples collected in 1996 and 1997 showed a decrease in 15 of the 24 sampling points. When the comparison was carried out in relation to each of the four main wind directions in the area, the highest decrease in PCDD/F concentrations (64%) corresponded to samples in the SE direction, while those in the NE direction showed also a notable reduction in total I-TEQ (44%). In contrast, PCDD/F levels in vegetation samples from NW and SW directions were increased (37% and 6%, respectively). When the data were evaluated according to the distance of the sampling points to the plant, the highest decrease in total I TEQ was found at 500 m from the stack. The results of this study seem to provide evidence for a decline in atmospheric emissions of PCDD/Fs in the area of Tarragona. PMID- 9753792 TI - Impact of safe water for drinking and cooking on five arsenic-affected families for 2 years in West Bengal, India. AB - The groundwater in seven districts of West Bengal, India, covering an area of 37,000 km2 with a population of 34 million, has been contaminated with arsenic. In 830 villages/wards more than 1.5 million people, out of the total population, drink the arsenic-contaminated water. Safe water from a source having < 0.002 mg 1(-1) arsenic has been supplied for 2 years to five affected families comprising 17 members (eight of them with arsenical skin-lesions) of different age groups for impact assessment study in terms of loss of arsenic through urine, hair and nail. The study indicates random observable fluctuations of arsenic concentration in urine among members on different scheduled sampling days with a declining trend, particularly during the first 6 months. Furthermore, the investigation showed that despite having safe water for drinking and cooking, the study group could not avoid an intake of arsenic, time and again, through edible herbs grown in contaminated water, food materials contaminated through washing, and the occasional drinking of contaminated water. After minimizing the level of contamination, a noteworthy declining trend after 8 months was observed in urine, hair and nails in all the cases, but not to that level observed in a normal population, due to prevailing elevated background level of arsenic in the area. The eight members, who had already developed skin lesions, are far from recovering completely, indicating a long-lasting damage. Statistical interpretation of the data are considered. PMID- 9753793 TI - Dietary intake of trace elements and polycyclic aromatic hydrocarbons via vegetables grown in an industrial Greek area. AB - In order to assess the importance of vegetables cultivated in industrialized regions in Greece as a dietary factor, the daily intake of trace elements and PAHs via vegetables were estimated. Intake estimations were based on vegetable availability data and analyses of vegetable contaminants. The mean daily intake of potentially toxic elements ranged between 1.7% (for As) and 23.6% (for Pb) the provisional tolerable daily intakes for adults. Vegetables were found to contribute significantly to the recommended daily intake of essential elements, such as Cr and Mn. The intakes of Cr, Pb, Zn, Co and Hg were highest in spring, whereas the intakes of As and Se were highest in winter. The daily intake of PAHs via vegetables was in general low. The potential doses of carcinogenic PAHs was at the lower range of estimates worldwide. PMID- 9753794 TI - The implementation of Agenda 21 'our common failure'? AB - Sustainable development has for a long time been considered the starting-point for environmental policy, with Agenda 21 leading the way to such development. Social and economic equity, as well as global efficiency and development, are considered among the requirements for sustainable development. Only when such development is reached, will the solution to environmental problems come into perspective. Although we are now 5 years along the road marked out by Agenda 21, the perspectives for achieving the objectives stated are still slight. Despite economic growth, global inequity is on the increase. The conversion of natural areas to agricultural land will greatly increase due to the population growth, especially in Asia and Africa. In North America and Europe this trend seems reversed: life expectancy is longer and people's health has improved. The developing world is catching up. However, there is more needed to bring the world population's welfare and social security levels up to par. At the same time the environmental pressures on the world's natural areas will have to be brought down substantially. This can be done using market-based instruments, supplemented where necessary with orders, bans, fiscal measures and agreements at national and international levels. Internationally, a policy directed to investments and trade is needed to stimulate developing regions to undertake technological renewal and work away backlogs. The technological potential is still gigantic, but investment in the social system -- in other words, in people -- will be necessary if we are actually to use this potential. PMID- 9753795 TI - Seasonality of heartworm infection and implications for chemoprophylaxis. AB - The current emphasis on heartworm prevention reflects the dependable protection provided by the monthly administered macrolide endectocides. This article reviews the prerequisites for heartworm transmission and the importance of daily temperature as a limiting factor in determining the seasonality of the transmission period. The practice of some veterinarians to continuously prescribe monthly chemoprophylaxis exaggerates the actual risk of heartworm transmission in most parts of the country and unnecessarily increases the cost of protection to their clients. Guidelines are provided for making an objective, conservative estimate of the earliest and latest dates for administering monthly chemoprophylaxis; and the use of seasonal projections for other clinical applications such as timing and interpretation of heartworm testing are discussed. PMID- 9753796 TI - The serologic diagnosis of heartworm infection in dogs and cats. PMID- 9753797 TI - Canine and feline caval syndrome. AB - The caval syndrome is a serious complication of chronic heartworm (Dirofilaria immitis) disease in dogs and cats. The syndrome is characterized by acute anorexia, respiratory distress, weakness, right-sided cardiac murmur, anemia, hemoglobinuria, hepatic and renal dysfunction, signs of forward and backward heart failure, and, possibly, disseminated intravascular coagulation (DIC). Retrograde migration of adult heartworms from the pulmonary arteries to the right ventricle, right atrium, and venae cavae causes disruption of the tricuspid apparatus. Valvular insufficiency, with concurrent pulmonary hypertension, reduces cardiac output thus resulting in forward and backward heart failure. Additionally, red blood cells are traumatized and hemolyzed as they flow through the mass of worms. Therapy consists of supportive care and the removal of the heartworm mass from the right ventricular inflow tract. Caval syndrome in dogs and cats is associated with high mortality rates and generally has a guarded to poor prognosis. PMID- 9753798 TI - Human dirofilariasis. AB - Although canine dirofilariasis has been recognized for over 300 years human dirofilariasis has received relatively little attention. Although human beings are dead-end hosts they can become infected and develop lesions associated with infection. Although these lesions are typically benign they may be misdiagnosed as more important disease and prompt unnecessary diagnostic procedures with attendant cost, discomfort and morbidity. In heavily endemic areas human dirofilariasis should be considered as a differential diagnosis for solitary pulmonary "coin" lesions. More widespread serologic screening and epidemiologic surveys are necessary to ascertain the importance of human dirofilariasis. PMID- 9753799 TI - Feline dirofilariasis. AB - Prevalence studies indicate that cats throughout the world residing in areas enzootic for heartworm disease in dogs are prone to infection. The prevalence in cats seems to parallel the prevalence in dogs in the same area but at a much lower level with cats having approximately one tenth (1/10) the prevalence seen in the unprotected dog population. This relative prevalence varies dramatically by region. This variation may reflect different vector populations with relative prevalence in cats being higher in regions with vectors which are indescriminant feeders. Indescriminant feeding vectors are those which will take a blood meal from any source as opposed to many mosquito species which have very specific host preferences. Aberrant migration of larvae seems to be much more common in the cat than in the dog. PMID- 9753800 TI - Dirofilariasis in the domestic ferret. AB - The popularity of pet ferrets in heartworm-endemic and -nonendemic areas is growing, with ferret ownership in the United States currently exceeding 10 million. The domestic ferret (Mustela putorius furo) has been reported to be susceptible to naturally-acquired and experimentally-induced infections of Dirofilaria immitis. Host-parasite relationships between D. immitis and domestic dogs and cats have been well studied, but there have been relatively few reports on infections in ferrets. Laboratory studies have shown the ferret to be highly susceptible, with infection and recovery rates similar to those achieved in the dog and higher than those seen in cats. Microfilaremia is characteristically of low concentration and transient in nature, similar to that seen in heartworm infected cats. A definitive diagnosis can be made from ELISA-based antigen tests, echocardiography, and angiography, but suggestive radiographic findings require additional supportive information to confirm a tentative diagnosis. Prevention has been shown to be effective with currently used canine prophylactic pharmaceutics, but effective treatment of adult heartworms in ferrets has not yet been confirmed by controlled studies. There is currently no approved drug for prevention or treatment of D. immitis in ferrets. PMID- 9753801 TI - Canine heartworm disease. PMID- 9753802 TI - Neuromuscular oropharyngeal dysphagia secondary to bone metastases. AB - Oropharyngeal dysphagia in adults is secondary to either a structural lesion or neuromuscular disorder of the upper esophageal sphincter. In cricopharyngeal achalasia (incomplete relaxation of the upper esophageal sphincter), the etiology is usually either related to neck surgery or other neuromuscular disorders. We report on a rare case of neuromuscular oropharyngeal dysphagia secondary to bone metastases to the base of the skull. The patient is an 81-year old man with prostate cancer with metastases to the sacrum. A gastroscopy was attempted to discern the etiology of his dysphagia, but the endoscope could not be advanced. A barium swollow showed cricopharyngeal achalasia, and an magnetic resonance image of the brain demonstrated bone destruction to the floor of the left posterior fossa in the region of the jugular foramen and foramen magnum. The bone destruction caused disruption of the glosso-pharyngeal and vagus nerves. Selective radiotherapy resulted in rapid improvement in his symptoms. The primary treatment of cricopharyngeal achalasia is to correct the underlying process, if possible. This case illustrates an unusual presentation of secondary cricopharyngeal achalasia caused by cranial nerve involvement secondary to bone metastases. PMID- 9753803 TI - Mental retardation and decision making: balancing autonomy and protection. AB - Balancing autonomy with protection in caring for patients with mental retardation remains a formidable task for many clinicians. Though historical debate has resulted in an attitude supporting increased autonomy for all patients generally, and in legislation for enhanced decision making for the developmentally disabled specifically, the operationalization of such attitudes and policies lacks sufficient attention in the literature. The authors discuss three important areas of decision making as these relate to the care of patients with mental retardation: competence, respect, and multiple stakeholders; and, offer recommendations in each area to provide clinicians with some guidance in balancing the goals of autonomy and protection in clinical care when treating people with mental retardation. PMID- 9753804 TI - Pain control following elective gastrointestinal surgery: is epidural anesthesia warranted? AB - Fifty-nine patients undergoing elective major gastrointestinal surgery were entered into a prospective, randomized trial between January 1993 and July 1994 comparing the effectiveness, side effects, and hospital costs of postoperative epidural anesthesia (Group 1, n = 29) and intramuscular narcotic injections (Group 2, n = 30). Epidural catheters were inserted by a team that supervised catheter care and infusion rates in the postoperative period. The nonepidural group received intramuscular injections on a regular basis. Patients filled out visual analog scales to measure levels of pain ( 1 = minimal, 10 = maximal) every eight hours. Patient activity, bowel, and urinary function were recorded by the nursing staff. Control of pain (as measured by the daily average visual analog score) was more effective in Group 1 (P < .001) on postoperative days 1-3 (1.3 vs 3.6 on day 1, 0.7 vs 2.6 on day 2, 0.9 vs 3 on day 3). There was no significant difference in mean values between groups 1 and 2 with respect to first ambulation on the hospital ward, onset of liquid diet, intake of solid food, first spontaneous voiding, first bowel movement, length of hospitalization, or charge of hospitalization ($13,439 +/- 7,452 vs $11,821 +/- 6,630). We conclude that epidural anesthesia significantly lessens incisional pain following major elective lower gastrointestinal surgery when compared to analgesic injections alone. However, while not statistically significant, the overall charge was increased by 14% in the epidural group. This finding should be examined in light of the relatively low pain level in patients receiving narcotic injections alone. PMID- 9753805 TI - Trovafloxacin: fourth generation fluoroquinolone. PMID- 9753806 TI - Why congress must regulate managed care. PMID- 9753807 TI - More on a sustainable medicine. PMID- 9753808 TI - "Physician assisted living". PMID- 9753809 TI - Be not afraid! PMID- 9753810 TI - The changing nature of conflict and famine vulnerability: the case of livestock raiding in Turkana District, Kenya. AB - The context of famine in Turkana has changed in recent years as the role played by livestock raiding in contributing to famine has increased. External responses to famine in Turkana have largely been drought driven, for example, food assistance and livestock restocking programmers, which have failed to meet the real needs of herders. The role of armed conflict in the form of raiding has been overlooked as a common feature of societies facing famine and food insecurity. The traditional livelihood-enhancing functions of livestock raiding are contrasted with the more predatory forms common today. The direct impact of raiding on livelihood security can be devastating, while the threat of raids and measures taken to cope with this uncertainty undermine herders' livelihood strategies. Self-imposed restrictions on mobility negatively affect the vegetation of both grazed and ungrazed pastures and restrict the available survival strategies. Predatory raiding leads to a collapse in the moral economy. Some implications of this for relief and development policy are considered, including approaches to conflict resolution. PMID- 9753811 TI - Rehabilitation in complex political emergencies: is rebuilding civil society the answer? AB - The paper examines the challenge of rehabilitation from complex political emergencies (CPEs) and identifies a strategy that is characterized as a civil society rebuilding approach. It focuses on Somalia and a case study of CARE project that aims to build the capacity of local NGOs. The paper argues that civil society in CPEs is simultaneously being undermined and contested by warring parties and emerging after state collapse. The scope of the paper is limited to one case study and that case study examines only a single aspect of civil society: national and international NGOs The paper therefore presents tentative and preliminary results based on limited research. However, in reviewing the literature and presenting a way of approaching the subject, it aims to suggest a starting-point for developing a theoretical framework for such research. The paper finds that international agencies have tended to focus on civil society institutions simply as conduits for aid money and that this has tended to create organisations which lack downward accountability, are dependent on donors and are not addressing the wider roles for civil society envisaged in the approach. Rebuilding civil society does hold out the promise of giving non-military interests a stronger voice and starting a process of changing the aid delivery culture. Achieving these objectives, however, will be a slow and largely indigenous process and there is a need for lowered expectations about what outside assistance can achieve. PMID- 9753812 TI - Injuries as a result of California earthquakes in the past decade. AB - The devastating effects of earthquakes have been demonstrated repeatedly in the past decade, through moderate and major earthquakes such as the October 1987 Whittier Narrows earthquake (5.9 on the Richter scale), the October 1989 Loma Prieta earthquake (7.1) and the January 1994 Northridge earthquake (6.7). While 'official' tallies of injuries and deaths are reported for each event, the numbers vary from report to report. For Northridge, the number of injuries vary between 8,000 and 12,000; the number of deaths from 33 to 73 (Peek-Asa et al., 1997; Durkin, 1996). While official estimates are commonly reported following disasters, the study of actual numbers, types and causes of casualties has not developed. In this paper, we identify the numbers and risk factors for injuries within community-based samples across three earthquakes in urban California. We first report the numbers and types of injuries in each earthquake and then identify risk factors specifically associated with the Northridge earthquake. PMID- 9753813 TI - Women, health and humanitarian aid in conflict. AB - The burden of political conflict on civilian populations has increased significantly over the last few decades. Increasingly, the provision of resources and services to these populations is coming under scrutiny; we highlight here the limited attention to gender in their provision. Women and men have different exposures to situations that affect health and access to health-care and have differential power to influence decisions regarding the provision of health services. We argue that the role of women in planning is central to the provision of effective, efficient and sensitive health-care to conflict-affected populations. PMID- 9753814 TI - The European Community Humanitarian Office (ECHO): 1992-1999 and beyond. AB - The European Community Humanitarian Office (ECHO) was established in 1992 to co ordinate and administer European Union (EU) humanitarian aid and by 1994 had grown to be the world's largest humanitarian aid donor. Since it was set up for an initial period of seven years, ECHO's performance and the question of its continuing existence will be reviewed next year; it seems unlikely to be abolished. In light of this, this paper starts with a description of how ECHO functions: its mandate, sources of finance and funding mechanisms. It looks then at some of the ongoing challenges that ECHO faces--its relations with Member States, its links to other Commission services and its attempts to establish a distinctive role within the humanitarian aid field. In terms of this last issue, the paper concludes that ECHO should focus instead on establishing itself as a credible donor agency. PMID- 9753815 TI - Satellite observations of Lava Lake activity at Nyiragongo volcano, ex-Zaire, during the Rwandan refugee crisis. AB - In June 1994 the summit crater of Nyiragongo volcano, located in the Great Lakes region of central Africa, began to fill with new lava, ending nearly 12 years of quiescence. An earlier eruption of the volcano in 1977 had culminated in the catastrophic draining of a lava lake through fissures in the crater wall, feeding highly mobile lava flows which reached the outskirts of Goma and killed more than 70 people. By July 1994, as many as 20,000 Hutu refugees were arriving in Goma every hour, only 18 km south from the summit of Nyiragongo. The exodus brought more than one million people to the camps near the town raising fears of a repeat of the 1977 eruption. This paper examines the role that satellite remote sensing could have played in surveillance of the volcano during this time, and demonstrates the potential for monitoring this and other volcanoes in the future. Images recorded by the spaceborne Advanced Very High Resolution Radiometer (AVHRR)--freely available over the Internet--provide semi-quantitative information on the activity of the volcano. The aim of this paper is to promote the wider use of readily available technologies. PMID- 9753816 TI - Biomechanical effects of rapid maxillary expansion on the craniofacial skeleton, studied by the finite element method. AB - The aim of this study was to evaluate the biomechanical effect of rapid maxillary expansion (RME) on the craniofacial complex by using a three-dimensional finite element model (FEM) of the craniofacial skeleton. The construction of the three dimensional FEM was based on computer tomography (CT) scans of the skull of a 12 year-old male subject. The CT pictures were digitized and converted to the finite element model by means of a procedure developed for the present study. The final mesh consisted of 2270 thick shell elements with 2120 nodes. The mechanical response in terms of displacement and von Mises stresses was determined by expanding the maxilla up to 5 mm on both sides. Viewed occlusally, the two halves of the maxilla were separated almost in a parallel manner during 1-, 3- and 5-mm expansions. The greatest widening was observed in the dento-alveolar areas, and gradually decreased through the superior structures. The width of the nasal cavity at the floor of the nose increased markedly. However, the postero-superior part of the nasal cavity was moved slightly medially. No displacement was observed in the parietal, frontal and occipital bones. High stress levels were observed in the canine and molar regions of the maxilla, lateral wall of the inferior nasal cavity, zygomatic and nasal bones, with the highest stress concentration at the pterygoid plates of the sphenoid bone in the region close to the cranial base. PMID- 9753817 TI - Enhanced angiogenesis induced by diffusible angiogenic growth factors released from human dental pulp explants of orthodontically moved teeth. AB - The aim of this study was to determine if diffusible angiogenic growth factors were released in human dental pulp during orthodontic tooth movement. These factors, if diffusible, could induce angiogenesis in other tissues, and may then be isolated and identified. The pulps from 14 premolar teeth treated with straight wire fixed orthodontic appliances for 2 weeks were compared with those of 14 untreated control premolar teeth from the same subjects. Following tooth extraction and sectioning, 1-mm horizontal sections of pulp tissue were embedded in collagen with 1-mm sections of rat aorta and co-cultured in growth media for up to 4 weeks. Sections of rat aorta alone were also cultured. Angiogenic changes in the form of microvessel growth were observed by light microscopy. Microvessel identification was confirmed by electron microscopy and by immunohistochemistry using staining for factor VIII-related antigen marker for endothelial cells. When compared at days 5, 10, and 14 of co-culture, the number of microvessels was significantly greater in the pulps from orthodontically moved teeth than in those from the control teeth. The number of rat aorta microvessels was also significantly greater when co-cultured with pulp from orthodontically moved teeth than with pulp from control teeth and when compared with control cultures of rat aorta alone. There were no significant differences in microvessel numbers between the rat aorta co-cultured with pulp from control teeth and control cultures of rat aorta alone. These results indicate an increase in angiogenic growth factors in the pulp of orthodontically moved teeth, and the enhanced response of the rat aorta when co-cultured with this pulp shows that these factors appear to be diffusible. PMID- 9753819 TI - Temporomandibular joint growth adaptation in Herbst treatment: a prospective magnetic resonance imaging and cephalometric roentgenographic study. AB - The aim of this investigation was to analyze three possible adaptive TMJ growth processes contributing to the increase in mandibular prognathism accomplished by Herbst appliance therapy: (1) condylar remodeling; (2) glenoid fossa remodeling; and (3) condyle-fossa relationship changes. The subjects were 15 consecutive Class II malocclusions (11 males and four females, aged 11.5-17.5 years) treated with the Herbst appliance for an average period of 7 months. Condylar remodelling, glenoid fossa remodelling, and condyle-fossa relationship changes were analyzed by means of magnetic resonance imaging (MRI). From each subject, four MR images were evaluated: before treatment, start of treatment (when the Herbst appliance was placed), during treatment (6-12 weeks after appliance placement), and after treatment (when the appliance was removed). 'Effective condylar growth' (= the sum of condylar remodelling, fossa remodelling, and condyle-fossa relationship changes) was analyzed with the aid of pre- and post treatment lateral cephalometric roentgenograms. In all 15 subjects, Herbst therapy resulted in an increase in mandibular prognathism. After 6-12 weeks of treatment MRI-signs of condylar remodelling were seen at the posterior-superior border in 29 of the 30 condyles. MRI-signs of glenoid fossa remodelling at the anterior surface of the postglenoid spine were noted in 22 of the joints. Condylar remodelling seemed to precede fossa remodelling. The condyle-fossa relationship was, on average unaffected by Herbst therapy. 'Effective condylar growth' during treatment was, on average, approximately five times larger in the Herbst group than in an untreated group with ideal occlusion (Bolton Standards) and the direction of the growth changes was relatively more horizontal in the treated cases. The results indicate that condylar as well as glenoid fossa remodelling seem to contribute significantly to the increase in mandibular prognathism resulting from Herbst treatment, while condyle-fossa relationship changes are of less importance. MRI renders an excellent opportunity to visualize temporomandibular remodelling growth processes. PMID- 9753818 TI - Bone scintigraphy of human temporomandibular joints during Herbst treatment: a case report. AB - Bone scintigraphy results were compared with changes shown on orthopantomographic radiographs in a patient with facial asymmetry, before, during, and after Herbst treatment, and followed up with control of growth activity in the temporomandibular joints (TMJs) after long-term retention. The present study showed that new bone formation (modelling) was initiated asymmetrically in TMJs during treatment. The results indicate that late development of right/left asymmetry in the occlusion can be corrected and normalized using the Herbst appliance therapy, stimulating a differentiated 'catch up' growth (modelling) in the TMJ with condyles. After treatment, original growth with asymmetric activity in the TMJ was re-established. This growth activity may re-establish the asymmetry in the sagittal occlusion and the face of the patient. It is therefore recommended that the occlusion should be maintained with an appliance which stabilizes the occlusion until cessation of the primary, endochondral growth. PMID- 9753820 TI - The initial effects of the treatment of Class II, division 1 malocclusions with the van Beek activator compared with the effects of the Herren activator and an activator-headgear combination. AB - The effects of the van Beek activator in the correction of Class II, division 1 malocclusions were studied in 39 children, aged 9-13 years (median 11 years), and compared with the effects of treatment with an activator according to Herren and with those of an activator-headgear combination. Profile cephalograms were made before treatment and at the attainment of a Class I molar relationship (median observation time 9 months). The median improvement of the overjet was 4.7 mm and of the molar relationship 3.6 mm. This was largely achieved skeletally by in increase in mandibular prognathism while the skeletal effect on the maxilla was clinically insignificant. The maxillary incisors retroclined and the mandibular incisors proclined moderately. In general, no intrusion of the maxillary incisors was found and the eruption of the molars could not be stopped. The effects differed partly between the sexes, with a larger mandibular skeletal and molar reaction in the boys and a larger maxillary molar movement in girls. The larger mandibular reaction in the boys might have been due to the on average 4.5 months longer treatment time. The skeletal effects of the treatment were similar with all three activator types. The control of the incisors was, however, superior with the van Beek activator, especially when compared with the Herren activator. PMID- 9753821 TI - Assessment of dental and facial aesthetics in adolescents. AB - The Index of Orthodontic Treatment Need (IOTN) is currently widely used for clinical, as well as epidemiological purposes. The Aesthetic Component (AC) of this index focuses on dental aesthetics and does not include facial aesthetics. The aim of the present study was to evaluate whether dental aesthetics as measured by the AC of the IOTN correlates with facial aesthetics. Facial attractiveness of 69 males and 75 females was scored on facial photographs at two different ages (11-13 years and 14-16 years). Scoring of the AC of the IOTN was undertaken on the dental casts. Increments between the observations at the two ages were calculated. To assess the association between scores of dental and facial aesthetics, correlation coefficients were calculated. There was a highly significant influence of orthodontic treatment on facial and dental aesthetic scores in the group which was not treated orthodontically at the first observation, but was treated orthodontically at the second observation. No correlation, however, was found between the increments in the facial aesthetic score and those in the dental aesthetic score. The results indicate that facial and dental aesthetics are influenced by different factors, and both should be evaluated when judging dentofacial aesthetics. PMID- 9753822 TI - Psychological aspects of cleft lip and palate. AB - In addition to the influences of family dynamics, educational and vocational factors on the social development and rehabilitation of CLP patients, psychological problems, such as lowered self-esteem and difficulties during social interaction, are also experienced by CLP individuals. As only 20 per cent of cleft teams world-wide carry out a psychological assessment for their patients, it is likely that the prevalence of psychological problems is higher than the literature suggests. To maximize the chances of a positive outcome in the care of cleft affected individuals, CLP patients who are concerned about their appearance or who experience psychosocial problems need to be identified by cleft teams. Interventions, such as counselling or social interaction skills training, should be offered in order that the patient's self-esteem and social self-confidence can be increased. Current research surrounding patient and parent satisfaction with cleft care suffers from several areas of methodological weakness. PMID- 9753823 TI - Short stature of prenatal origin: craniofacial growth and dental maturation. AB - Recently, children born small for gestational age (SGA) with a catch-up growth failure, have been selected for high dose growth hormone (GH) treatment. In order to gain greater insight concerning dentofacial growth and maturation of these patients, and to evaluate the possible effects of high does GH administration on facial structures, craniofacial growth and dental maturation were evaluated in short SGA persons. Seventy-seven cephalograms and orthopantomograms were available from 48 subjects, aged between 2 and 32 years. Craniofacial growth was assessed by calculating age- and gender-specific standard deviation scores (SDS) for eight linear and five angular measurements. Tooth formation was evaluated by means of a dental delay score (i.e. dental age minus chronological age). The SDS for craniofacial growth measurements for the lateral aspect showed a short anterior cranial base (-1.8 SDS), a small retropositioned mandible (< or = -1.7 SDS) and a small maxilla (-1.5 SDS); a high mandibular plane angle (+1.9 SDS) and a wide cranial base angle (+1 SDS). These findings result in a small retrognathic face with a relatively increased lower anterior face height (+1.7 SDS). In contrast to skeletal maturation, dental age was not delayed. The general growth retardation is, apparently, reflected to a differential extent within the craniofacial complex, while dental maturation appears to be a distinct process tightly linked to chronological age, and independent of general growth and bone age. PMID- 9753824 TI - Apical root resorption during orthodontic treatment of patients with multiple aplasia: a study of maxillary incisors. AB - The aim of this study was to evaluate the risk of root resorption during orthodontic treatment of patients with aplasia, and to analyse the relative importance of some anamnestic and treatment variables. The subjects comprised 68 orthodontically treated patients with 1-16 congenitally missing teeth. The age of the patients was 11-20 years (mean 15 years). All patients were treated with fixed edgewise appliances. The purpose of the orthodontic treatment varied: to create optimal conditions for prosthetic restorations or osseointegrated implants, or to achieve aesthetic and functional alignment of teeth in less severe cases. The degree of root resorption was assessed before and after treatment from intra-oral radiographs of the maxillary incisors using a scale of 0-4. In all, 186 maxillary incisors were evaluated. The degree of apical root resorption was significantly greater in cases of multiple aplasia (4-16 missing teeth) than in those with only one to three missing teeth. Root form, treatment time with rectangular wires and intermaxillary elastics, and total treatment time were significantly related to root resorption. Discriminant analysis disclosed that the following variables were the most important determinants of root resorption: number of missing teeth, root form, and time with rectangular archwires and intermaxillary elastics. It is concluded that there is a high risk of apical root resorption during orthodontic treatment in patients with multiple aplasia (four or more teeth), in particular in teeth with an abnormal root form and lengthy treatment with elastics and rectangular archwires. PMID- 9753825 TI - Changes in proteoglycan and collagen content in the mandibular condylar cartilage of the rabbit caused by an altered relationship between the condyle and glenoid fossa. AB - Twenty 5-day-old New Zealand rabbits underwent surgery to induce premature synostosis of the cranial sutures, resulting in posterior displacement of the glenoid fossa. Twenty sham-operated rabbits served as controls. The animals were killed at age 15 days for histochemical and biochemical analyses. The collagen content of the superior region of the condyle determined biochemically was lower in treated animals than in controls. Biochemical and histochemical analyses revealed the proteoglycan content to be significantly reduced in the superior region of the condyle (P < or = 0.001). Low levels of aggregating proteoglycans were seen. Since levels of aggregating proteoglycans decreased, catabolism must have exceeded their synthesis or the monomers must have been unable to aggregate and escaped from the tissue. It is concluded that an experiment in which the location of the mandibular condyle in the glenoid fossa is changed, while causing marked reductions in amounts of both collagen and proteoglycans in the cartilage tissue of the mandibular condyle, will also induce changes resembling those observed in animal models of arthritis. It is possible that the two phenomena have similar mechanisms. PMID- 9753826 TI - The sella turcica in children with lumbosacral myelomeningocele. AB - The purpose of the present study was to analyze the morphology of the sella turcica in children born with myelomeningocele. Profile radiographs from 16 children (nine females and seven males) born with myelomeningocele were analysed. The contour of the anterior wall of the sella turcica in myelomeningocele patients, instead of following the normal cranio-caudal direction, was always in an obliquely antero-posterior direction. The sella turcica thus appeared broad cranially with a diverging anterior wall, or with both diverging anterior and posterior walls. This appearance gave and impression of a wide sella turcica in myelomeningocele with less depth than normal. The investigation has drawn attention to the fact that congenital malformations in the axial skeleton, even though, as in the case of myelomeningocele, they are located far from the cranial base, may have manifested themselves in the cranial base as well. The pathogenetic relationship between these manifestations is to be found in the early embryonic structure, the notochord. With the concept of embryological developmental fields, defined as areas with a common developmental origin, such as the notochordal field involved in myelomeningocele, new ways seem to be emerging for an improvement of aetiologically based diagnosis and treatment. PMID- 9753827 TI - Cephalometric analysis of changes in occlusal relationship. AB - One of the main problems in assessing the mode of action of various treatment modalities is the method of measuring the treatment change. The purpose of the present study was to develop a cephalometric method that would permit a detailed evaluation of the individual growth processes (skeletal and dental) that contribute to the overall change in occlusal relationship. The change in molar relationship was resolved in five components, i.e. translation of the maxilla, of the upper molar, of the mandible, of the lower molar, and rotation of the mandible. These movements were recorded using regional superimposition of various structures, with the aid of a computer program. Derotation of the mandible was performed to remove any confounding effects of total mandibular rotation on the interpretation of the measurements. The results show that assessment of treatment effects can be carried out by comparison of the five resulting vectors. PMID- 9753828 TI - Comments about physician assisted suicide. PMID- 9753829 TI - UH med schools plan. PMID- 9753830 TI - Chronic meningococcemia mimicking acute rheumatic fever. PMID- 9753831 TI - Epidemiology of congenital diaphragmatic hernia, Hawaii, 1987-1996. AB - Congenital diaphragmatic hernias (CDHs) in Hawaii between 1987 and 1996 were examined with data from a birth defects surveillance system. There were 51 cases of CDH (prevalence 2.45 per 10,000 births). Forty-nine percent of livebirths survived, an increase over the rate reported in Hawaii in 1975-1982. These results are similar to those reported by other population-based studies. PMID- 9753832 TI - Noncontraceptive health benefits of the oral contraceptive pill. PMID- 9753833 TI - Who is a nurse? PMID- 9753834 TI - Interview with Faye G. Abdellah on nursing research and health policy. PMID- 9753835 TI - Cost-effectiveness analysis and nursing research--is there a fit? PMID- 9753836 TI - Milton Weinstein's insights on the development, use, and methodologic problems in cost-effectiveness analysis. PMID- 9753837 TI - Methods for conducting and reporting cost-effectiveness analysis in nursing. PMID- 9753838 TI - Cost-effectiveness analysis in the nursing literature, 1992-1996. AB - PURPOSE: To review the recommendations by the U.S. Panel on Cost-Effectiveness in Health and Medicine (panel) for use in future nursing research. The review (a) provides a critique of the nursing cost-effectiveness and cost utility literature from the perspective of the recommendations set forth by the panel and other recognized authorities in cost-effectiveness analysis (CEA), (b) constructs an interdisciplinary framework to show the steps in the conduct of CEA, (c) makes the techniques and major findings of nursing CEA studies available and understandable, and (d) offers guidelines for the incorporation of CEA into the evaluation of future nursing intervention and research. DATA SOURCES: Seven nursing studies published between 1992 and 1996 that compared two or more interventions for costs and outcomes. ORGANIZING FRAMEWORK: For each study, the (a) perspective, (b) net costs, (c) net effect, (d) analysis of costs and effects, and (e) decision outcomes were analyzed. FINDINGS: If the panel's recommendations reflect the problems in the health care CEA literature in general, then on balance, the nursing CEA 1992-1996 studies are no more or less flawed than CEA studies in the health or medical care fields. CONCLUSIONS: Methodologic guidelines and interdisciplinary strategies are needed to advance the progress of nursing cost-effectiveness research. PMID- 9753839 TI - Clinical validation of ineffective breathing pattern, ineffective airway clearance, and impaired gas exchange. AB - PURPOSE: To describe the clinical validation of symptoms or defining characteristics of three respiratory diagnoses. The contributing factors or etiologies of the diagnoses were identified and the degree of importance of 30 nursing interventions, 15 direct care and 15 teaching, was rated for each diagnosis and each patient. Three nursing diagnoses--ineffective breathing pattern (IBP), ineffective airway clearance (IAC), and impaired gas exchange (IGE)--were among the most frequently used, yet no reported clinical studies validated the defining characteristics of these diagnoses. This study answers the research questions: What are the defining characteristics of IBP, IAC, and IGE? What are the etiologies of IBP, IAC, AND IGE? What are the most important interventions for IBP, IAC, and IGE? DESIGN: Standardized clinical validation using a convenience sample of 76 people hospitalized with medical and surgical diagnoses, in one U.S. city, and identified as having one of the three diagnoses. Data were collected in 1992-1993. METHODS: A literature-based concept analysis generated 37 possible defining characteristics for the three diagnoses which were included in the instrument. The nurse experts conducted a health history and physical examination of each patients and decided (a) whether the 37 defining characteristics were present or absent, (b) the degree of importance of each possible defining characteristic for making one or more of the diagnoses, (c) the etiologies, and (d) which of the 30 nursing interventions were important for each diagnosis and patient. FINDINGS: For each diagnosis, many of the 37 possible defining characteristics were judged as present but few reached the criterion of .50 as important for making one of the diagnoses. Two of the possible defining characteristics reached this criterion for IBP, seven for IAC, and two for IGE. In contrast to the defining characteristics approved by NANDA, the subjective cues of "expresses fatigue" and "expresses anxiety" were judged as important for making one or more of the diagnoses. CONCLUSIONS: Clinical validation methods allow discriminating among defining characteristics. Data that are present are not necessarily characteristic of a diagnosis, and the subjective cues of expresses fatigue or anxiety may be important for making these diagnoses. PMID- 9753840 TI - Insights of nurses about assault in hospital-based emergency departments. AB - PURPOSE: To explore contributing factors, consequences, and solutions to the assault of nurses working in U.S. hospital emergency departments. This preliminary study targeted emergency nurses whose risk for assault was significantly greater than many other workers. Exploring nurses' opinions about factors they believe contribute to assault provides important information for designing acceptable preventive measures. DESIGN: In this descriptive study, an ecological, occupational-health framework was used which integrates personal, organizational, and societal influences. The six components of the framework were personal factors, workplace factors, environmental factors, assault injuries, solutions, and effects of workplace violence. METHOD: Four focus groups were used comprised of 22 RNs employed in emergency departments in one large metropolitan area in the United States. Half the nurses had been physically assaulted at work. FINDINGS: Fourteen themes emerged: nurse attitude, vulnerability, security, administrative issues, assault reporting, safety training, beyond control, societal changes, types of patients, geographic location, pervasiveness of anger, previous assault experiences, effects and possible solutions. Personal, workplace, and environmental factors all contribute to assault. Verbal and physical assaults are common and affect nurses' personal and professional lives. CONCLUSIONS: Assault-related injuries are preventable. Only physical injuries are treated; all employees who have been verbally or physically assaulted should be referred for post-incident debriefing. Hospital managers should implement violence-prevention programs. The ecological, occupational health framework is useful for identifying factors that contribute to assault. PMID- 9753841 TI - Comparison of continuing care communication. AB - PURPOSE: To describe and compare the patient-care communication exchanged between personnel in hospitals and nursing homes (NHs) and hospitals and home health agencies (HHAs) in referrals of elderly patients using an adaptation of classic communication theory. Little research on patient-care communication across organizational settings has been reported. This study offers baseline information about inter-organizational communication and insight into barriers to patient care communication. DESIGN: A retrospective, descriptive study using a convenience sample of 455 medical records of referrals to NHs and 300 to HHAs. METHODS: Medical records were audited and a Referral Data Inventory (with established reliability and validity) was completed for each of the records reviewed. Data were collected between January and June 1995. FINDINGS: Greater amounts of referral data were transferred to NHs, than to HHAs. Patient information was composed largely of background and medical data, followed by some nursing care data and limited psychosocial data. Hospitals employed more formal channels of communication in referring patients to NHs than to HHAs, and communicated information more promptly. Some organizational factors related to both the referring hospitals and receiving organizations resulted in discrepancies in patient-care communication. CONCLUSIONS: Continuity of patient care involves a series of coordinating linkages across time, settings, providers, and consumers of health care. Communication is a core task in coordinating patient care. Increased and improved inter-organizational communication is needed when patients are discharged to nursing homes or home health agencies. PMID- 9753842 TI - Translation and validation of nursing interventions classification (NIC) in English and Korean. AB - PURPOSE: To describe the translation and validation of the Nursing Interventions Classification (NIC) Use Questionnaire. METHODS: The NIC Use Questionnaire was translated to Korean using back-translation and three criteria to test the semantic, content, and technical equivalence of the original and translated version. FINDINGS: Three factors made translation difficult including structural differences in the two languages, long modifier phrases in English, and nonequivalent words in Korean. CONCLUSIONS: This translation facilitates communication among nurses. The findings suggest a method for similar validation in other languages. PMID- 9753843 TI - Change in the word symbol for nurse in Korea. AB - PURPOSE: To analyze the 1987 change in the written symbol for the work "nurse" in Korea, engendered by Mo Im Kim. DESIGN: Descriptive historical and narrative analysis. METHODS: Data were collected from documents in four libraries 1992-1996 in Seoul, Korea and Dallas, Texas. Kim and 20 of her friends, colleagues, and associates were interviewed. Findings from the interviews were compared with historical nursing documents and letters. FINDINGS: Kim's work with her colleagues and their joint endeavors resulted in a change in nursing's status and responsibilities through an alteration of the Korean symbol for nursing. The symbol change resulted in the definition of nursing moving from that of manual laborer to that of teacher or professional. CONCLUSIONS: Changing the symbol for nurse reflected the shift in nursing's responsibilities and caused a legal change in the official civil service classification of nurse. This, in turn, has changed the status of nursing and led to more professional practice. PMID- 9753844 TI - Health promotion among immigrant women from India living in Canada. AB - PURPOSE: To describe the health promoting practices of immigrant women from India and how cultural knowledge, norms, and values influence their behavior. DESIGN: Descriptive using a small convenience sample. METHOD: Using ethnographic methodology, 20 women between the ages of 40 and 70 years living in Canada, 1995 1996, were interviewed individually using open-ended questions. Community get togethers were also observed. FINDINGS: The sample of women consistently reported that to remain healthy, a good diet, activity, and weight control were important. In addition, prayers, spiritual activities, and good relationships with families helped. CONCLUSIONS: Health professionals should be aware of the special needs of immigrant women to help them promote healthy lifestyles in their cultural context. PMID- 9753845 TI - Spirituality and chronic illness. AB - PURPOSE: To present a comprehensive overview of spirituality and identify strategies to support the spiritual dimensions of nursing care for people with chronic illness, focusing specifically on HIV-related illness and AIDS. SIGNIFICANCE AND SCOPE: The AIDS crisis has brought new emphasis to the need to develop therapeutic interventions to support the coping resources of people living and dying with chronic illness. Conceptual, theoretical, and empirical knowledge related to spirituality was reviewed, integrated, and interpreted within the context of nursing care for this population, emphasizing the spiritual needs of people with HIV-related illness and AIDS. CONCLUSIONS AND IMPLICATIONS: Spirituality has evolved beyond religious considerations to encompass multidimensional and existential perspectives that are integral to maintaining well-being for the chronically ill. A deeper understanding of spirituality enhances the potential for nurses to identify spiritual needs and incorporate spiritual caring into practice. PMID- 9753846 TI - Plato, Nightingale, and contemporary nursing. AB - PURPOSE: To explore the influence of Plato's ideal of leadership on Nightingale's vision for nursing. Nightingale left her imprint on nursing; greater understanding of Nightingale's legacy clarifies several contemporary problems in the field. ORGANIZING FRAMEWORK: Qualitative, historical research in which the great person framework is used to focus on Nightingale and Plato. Nightingale's familiarity with Plato, whose work she read in the original Greek, is established. SOURCES: Evidence for Platonic influence is uncovered in primary sources, then inductive reasoning and reflective analysis are used. FINDINGS: Nightingale's conception of nursing is akin to Plato's conception of public service. Platonic education aims to turn talented people into leading citizens who care for the good of the community. Modern nursing did not evolve from a craft guild tradition, as did medicine. This profound difference in the constitution of nursing is part of what distinguishes it. CONCLUSIONS: Reminiscent of Plato's "Republic," Nightingale's legacy leaves a guardian-like stamp on nursing. Although this legacy also leaves nursing with some ambiguities about technical training and hierarchical systems, it is an orientation to the general good that provides the basis for nursing's holism. PMID- 9753847 TI - Echocardiography in prosthetic valve disease. PMID- 9753848 TI - Antiarrhythmic trials: a decade of growth. PMID- 9753849 TI - Risk factors for coronary artery disease and levels of lipoprotein(a) and fat soluble antioxidant vitamins in Asian Indians of USA. AB - High rates of coronary artery disease have been reported in the Asian Indians who have migrated to other countries. Although many coronary artery disease risk factors (diabetes, high serum cholesterol, lipoprotein[a], and smoking) have been suggested, studies of coronary artery disease risk factors in Asian Indians living in USA are only a few. We investigated coronary artery disease risk factors in 110 Asian Indian physicians living in USA by questionnaire and measurement of their serum lipids and fat-soluble antioxidant vitamins. Differences in risk factors between genders, vegetarian diets and diabetic status were also studied. We found that lipoprotein(a) (mean=20 mg/dl), low density lipoprotein cholesterol, and diabetes (prevalence of 7.5%) are more important risk factors for coronary artery disease, but not smoking, when compared to other Americans. Higher levels of low density lipoprotein cholesterol, retinol, alpha tocopherol, cryptoxanthin and lycopene, and lower levels of high density lipoprotein cholesterol were found in the males than in females. Comparable levels of lipoprotein(a) were found for males and females. Vegetarians, compared to non-vegetarians, had similar levels of lipids and fat-soluble antioxidants. Lower levels of retinol, lutein/zeaxanthin and lycopene were found in the diabetics compared to non-diabetics. These findings suggest that (1) the control of low density/high density lipoprotein cholesterol levels could be important in prevention of coronary artery disease in Indian males, (2) the vegetarian diets of Asian Indians do not favourably influence the serum lipid and antioxidant levels, and (3) increased serum levels of antioxidants may be beneficial for diabetics. Furthermore, for the first time, we show that serum levels of lutein/zeaxanthin and lycopene are significantly lower in the diabetics. PMID- 9753850 TI - Serum tocopherols and lipids in patients with coronary artery disease. AB - Serum tocopherols and lipids levels in 40 coronary artery disease patients were statistically analysed in relation to those of the normal controls and an attempt was made to define the correlation between tocopherols and various lipids. Serum tocopherols levels were found to be significantly low in coronary artery disease patients as compared to those of the controls implicating an inverse relationship between coronary artery disease and serum level of tocopherols. Coronary artery disease population did not register any significant change in various blood lipids, except triglyceride fraction, as compared to that of the normal population. Serum tocopherols and lipids were also analysed in relation to age and sex. The preliminary data suggest that coronary artery disease patients have sub-optimal tocopherols level (0.58 +/- 0.26 mg/dl) as compared to that of the normal controls (0.81 +/- 0.34 mg/dl; p < 0.001), which may be an important risk and/or contributing factor for atherogenesis and coronary artery disease PMID- 9753851 TI - Plasma homocysteine levels in patients with coronary heart disease. AB - Hyperhomocysteinemia is being identified as a risk factor for coronary heart disease but its role among Asian Indians has not been studied. This has practical importance because (1) the data generated in the West may not represent Indian population, and (2) the condition is remediable. To assess the magnitude of this problem, we studied 56 patients with coronary heart disease, and 53 control subjects. Details of diet, smoking, medication, hypertension and diabetes were recorded; lipids and sugar levels were estimated in all. Patients with renal and liver diseases were excluded. Serum homocysteine was estimated using liquid chromatography. Both the groups were comparable by age and sex. Higher, but statistically insignificant homocysteine levels were seen in patients with coronary heart disease: 10.98 +/- 9.04 nmol/ml vs 9.41 +/- 3.60 nmol/ml in control subjects. Among males, higher, but statistically insignificant levels were seen in coronary heart disease patients: 11.96 +/- 9.41 nmol/ml vs 9.87 +/- 3.50 nmol/ ml in control subjects; among females, the levels were lower though not significant: 5.10 +/- 1.64 nmol/ml vs 6.39 +/- 2.99 nmol/ml. Sub-group analysis with age 40 as dividing point did not show significant difference. Six (10.7%) patients with coronary heart disease and three (5.7%) control subjects had homocysteine levels above 95th percentile of control subjects (p = NS). Twenty-three (41.1%) coronary heart disease patients and 19 (35.9%) control subjects had levels above 10 nmol/ml (p = NS). We conclude that homocysteine is not a major risk factor for coronary heart disease in the study population. The lack of statistical significance could be due to inadequate sample size although some past studies reporting statistically significant association between coronary heart disease and homocysteine involved similar or smaller number of subjects. Larger studies are warranted to see if ethnic differences also have any role. PMID- 9753852 TI - Correlation of Braunwald's clinical classification of unstable angina pectoris with angiographic extent of disease, lesion morphology and intra-luminal thrombus. AB - One hundred consecutive patients (81 male and 19 female) with unstable angina pectoris undergoing coronary angiography were divided according to Braunwald's clinical classification. Seventeen (17%) patients had new onset angina (class I), 68 (68%) sub-acute angina (class II) and 15 (15%) had acute rest angina (class III). Twenty-seven (27%) patients had secondary unstable angina pectoris (class A), 49 (49%) primary unstable angina (class B) and 24 (24%) had post-infarction unstable angina (class C). ST-T wave changes on ECG were present in 54 (54%) while absent in 46 (46%) patients. On coronary angiography, 26 (26%) patients had single vessel disease, 30 (30%) double vessel disease and 39 (39%) patients had triple vessel disease. Five (5%) patients were found to have normal coronaries. Classification of patients according to Braunwald's clinical classification showed single vessel disease to be higher in class I as compared to class II (47% vs 22%; p = 0.04) and classes III (47% vs 20%; p<0.01). Single vessel disease was found to be higher in class C as compared to class B (41.7% vs 16.4; p = 0.01). Double vessel disease was higher in class B as compared to class A (40.8% vs 18.5%, p = 0.04). Triple vessel disease incidence was not found to be significantly different among different clinical classes. Morphology of coronary artery lesions was classified according to Ambrose's classification. Out of the total of 248 lesions in the whole study group, there were 68 (27.42%) concentric lesions, 55 (22.18%) eccentric type I lesions, 23 (9.27%) eccentric type II lesions, 42 (16.94%) multiple irregularity lesions and 60 (24.19%) totally occluded lesions. Concentric lesions were found to be higher in class C as compared to class B (40% vs 19.8%; p = 0.014). Statistically significant difference was not present in the distribution of other morphological type of lesions among different clinical classes. In the whole study group, intra-luminal thrombus was found to be present in 17 (17%) of patients. Distribution of intra luminal thrombus according to Braunwald's classification showed that none of the patients in class I had intra-luminal thrombus, while 13 (19.1%) patients in class II and 4(26.7%) in class III had intra-luminal thrombus. The difference in the occurrence of intra-luminal thrombus between class I and class II (p = 0.004) and class I and class III (p = 0 .03 was found to be significant. Thus, majority of patients undergoing coronary angiography had primary sub-acute rest angina. Single vessel disease was higher in new onset angina. Patients with unstable angina pectoris and ST-T changes on ECG had higher number of lesions per patient and higher eccentric type I lesions. Intra-luminal thrombus was more frequently encountered with acute rest angina. However, the distribution of different morphological type of lesions on coronary angiography did not differ significantly in different clinical classes of unstable angina pectoris divided according to Braunwald's classification. PMID- 9753853 TI - Elective stent implantation after optimal debulking for complex coronary lesions: acute and mid-term results. AB - Between January 1995 to December 1997, 45 patients with complex lesions in coronary arteries were treated by using the strategy of initial debulking with an atherectomy device followed by elective stenting. Their age ranged from 35-73 years (mean +/- SD:53.9 +/- 9.1) and 93.3 percent were males. The lesion morphology was type B1 in 14 (31.1%), B2 in 16 (35.6%) and type C in 13 (28.9%) patients. The choice of atherectomy device, based primarily on the morphology of lesion, was rotational atherectomy in 23 (51.1%) and directional coronary atherectomy in 22 (48.9%) patients. While majority (73.9%) of the lesions treated by rotablation were long, diffuse and calcified, directional atherectomy was preferred for highly eccentric stenoses in large-sized arteries. All patients underwent elective stent implantation after optimal lesion debulking using a mean burr size of 1.74 +/- 0.2mm for rotablation and a 7Fr. atherocath in majority (90.9%) of patients treated by directional coronary atherectomy. Angiographic success was achieved in all, while clinical success was 97.8 percent. One patient died of acute-on-chronic renal failure during hospitalisation. There were no other major in-hospital adverse cardiac events. At a median follow-up of 13 months (range 1-36 months), recurrence of angina developed in 10 (22.7%), out in which target lesion revascularisation was required in 5 (11.4%) and elective coronary artery bypass graft surgery in one (2.2%) patient. The event-free survival as calculated by the Kaplan-Meier method was 85.8 percent at six, 77.2 at 12 71.7 percent at 18 months of follow-up. In conclusion, optimal debulking before stent implantation provides a larger lumen, and thus eliminates sub-acute stent thrombosis in complex coronary lesions. This strategy also resulted in a high incidence of event-free survival and a low frequency of target lesion revascularisation on mid-term follow-up. PMID- 9753854 TI - Post-cardiotomy intra-aortic balloon counter pulsation: application and prognostic evaluation. AB - Cardiac assistance by intra-aortic balloon counter pulsation was studied in 113 cardiac surgical cases comprising 91 male and 22 female patients. This included 82 percent of patients having coronary artery bypass surgery, while 18 percent were operated for valvular lesions. It was observed that the time of institution of cardiac assistance by intra-aortic balloon counter pulsation, following cardiac surgery, was of prime importance to decrease patient mortality. It was lowest (16%) when the balloon was inserted for assistance before termination and highest (50%) when there was delay of more than 15 minutes following termination of cardiopulmonary bypass. Early balloon assistance significantly lowered the pulmonary capillary wedge pressure and usually 1:2 augmentation was more effective, probably because of existing tachycardia in most patients. Advances in catheter technology have reduced the vascular complication at the insertion site. Percutaneous insertion had less local complications (13.3%) than open arteriotomy technique (31.2%). Similarly with sheathless insertion, complications were less (6.6%) in comparison to sheathed insertion (21.7%). Proper placement of balloon avoided position-related complications and there was no compromise of blood flow through left internal mammary artery as noticed in our series. PMID- 9753855 TI - Surgery for aortic valve endocarditis. AB - From March 1994 to March 1997, 36 patients with aortic valve endocarditis were managed surgically. Of these, 30 patients had native valve endocarditis and six had prosthetic valve endocarditis. In patients with native valve endocarditis, surgical procedures included aortic valve repair (n=6), homograft aortic valve replacement (n=9), Ross procedure (n=5) and prosthetic aortic valve replacement (n=10). There were three early and two late deaths in this group. In patients with prosthetic valve endocarditis, aortic valve replacement with a homograft was performed in all. Active infection and prosthetic valve endocarditis were the most important predictors of early mortality. The availability of a homograft valve provides an alternative to prosthetic valve replacement in patients with aortic valve endocarditis. PMID- 9753856 TI - Pathology of the heart in acquired immunodeficiency syndrome. AB - The spectrum of cardiac lesions in patients with acquired immunodeficiency syndrome in India is not described. To determine the extent of involvement of the heart with this disease, an autopsy study of 52 subjects having acquired immunodeficiency syndrome was carried out. Multiple sections were obtained from different anatomical parts of each heart. Forty-eight of the 52 hearts showed subtle microscopic changes, the most common being myocardial atrophy (48 cases), lymphocytic pericarditis (38 cases), fibrinous pericarditis (1 case), pericardial fibrosis (1 case), lymphocytic myocarditis (29 cases) and myocardial fibrosis (7 cases). Cryptococcosis of the heart was noticed in two cases, while in one case toxoplasmic myocarditis was identified. In only one case clinical presentation of cardiac involvement (pericardial effusion) was noted, which indicates that in spite of the presence of significant pathology in the heart, overt cardiac manifestations are infrequently seen in patients with acquired immunodeficiency syndrome. PMID- 9753857 TI - The first ever radial artery as a conduit in femoro-popliteal bypass: a case report. PMID- 9753858 TI - Acute myocardial infarction in tetralogy of Fallot. PMID- 9753859 TI - Anomalous drainage of right superior vena cava into the left atrium. PMID- 9753860 TI - Absent left circumflex coronary artery. PMID- 9753861 TI - Isolated iliac artery aneurysm: treatment by percutaneous stent-graft placement. PMID- 9753863 TI - Rapid and reliable site directed mutagenesis using Kunkel's approach. AB - Oligonucleotide based site directed mutagenesis (SDM) is an invaluable technique in molecular biology. Among the various methods developed for SDM, the PCR-based approach, Kunkel's and the Eckstein's procedures are widely used. The Kunkel's method, on account of its cost effectiveness and simplicity, is preferred by many a scientist. However, a general drawback of this method is the high background due to persistence of the parent template resulting in low efficiency of mutagenesis. In this report, we describe a modification of the Kunkel's method to increase the efficiency of selecting against the wild type strand. We have used Sequenase for the extension reaction, and introduced an in vitro UDG step to enhance the biological selection against the parent strand. Consequently, the efficiency of the modified method is enhanced to allow screening of the mutants directly by DNA sequencing. A step by step single tube protocol which is over in less than three hours makes it a method of choice for efficient and cost effective site directed mutagenesis. PMID- 9753862 TI - Sinus venosus atrial septal defect with myasthenia gravis--a rare association. PMID- 9753864 TI - Insight into the environment of tryptophan in a hydrophobic model peptide upon aggregation and interaction with lipid vesicles: a steady state and time resolved fluorescence study. AB - Fluorescence spectroscopy is extensively used to monitor binding of peptides to lipid vesicles as well as orientation in the lipid bilayer. In steady-state fluorescence, the emission characteristics of intrinsic and extrinsic fluorophores, which are sensitive to environment are monitored. Life time measurements should yield useful information about the location and flexibility of fluorophores, as these factors have a significant effect on the life times. However, studies on protein structure and dynamics indicate that interpretation of life-time data is complicated (Beechem. J.M. and Brand, L. (1985) Annu. Rev. Biochem. 54, 43-71). Hence, simple well-defined systems should help in interpretation of life time data, especially in lipid-peptide interactions. In order to examine how fluorescence characteristics of tryptophan and anthroyl group would reflect molecular details of peptide aggregation and lipid-peptide interaction, studies have been carried out on a model hydrophobic peptide and its fatty acylated derivative. Steady-state fluorescence measurements suggest that: (1) the fatty acyl chain attached to an amino acid associates with the peptide chain in aqueous environment. (2) In the lipid bilayer, the acyl chain is oriented perpendicular to the lipid bilayer surface with the peptide chain at an angle to it. Analysis of the fluorescence decay of tryptophan indicates the predominance of a very short life-time component (<1ns) in aqueous environment and lipid-vesicles. Since the preexponentials were not negative, it is unlikely that this is due to extensive deactivation process. We attribute the observation of the low life time component to predominance of one rotamer around (C alpha-C beta)bond of tryptophan in aqueous and lipid environments. Our investigations suggest that fluorescence life time data need to be complemented with steady state measurements to get an insight into details of lipid-peptide interaction. PMID- 9753865 TI - Thermal stabilization of multimeric proteins: a case study with alpha-globulin. AB - Preferential interaction parameters of multisubunit protein, alpha-globulin and monomeric protein human serum albumin (HSA) were determined in different cosolvents using precision densitymetry. The apparent partial specific volumes were determined under both isomolal and isopotential conditions for alpha globulin in 0.02 M glycine-NaOH buffer at pH 10 and the values were 0.692+/-0.002 and 0.688+/-0.001, ml/g, respectively, at 20.00+/-0.01 degrees C. From the partial specific volume data with cosolvents the preferential interaction parameter (xi3) and other thermodynamic parameters were calculated at different solvent concentrations. The (xi3) values increased with an increase in the solvent concentration up to 30% and reached a maximum with the values of-0.111+/ 0.018 g/g and -0.076+/-0.012 g/g in sucrose and sorbitol, respectively. In glycerol the (xi3) values decreased with an increase in solvent concentration. The above data is further supported by thermal denaturation profiles in which the apparent thermal denaturation temperature (apparent Tm) of alpha-globulin shows an increase from 63 degrees C to higher temperatures in the order of sucrose, sorbitol and glycerol. Alpha-globulin showed coagulation due to protein interaction at temperatures above 50 degree C. The apparent Tm of 63 degrees C for control protein was increased significantly up to 75 degrees C in 40% sorbitol with two fold increase in the delta(S) values showing the increased structural stability of alpha-globulin. At high solvent concentration the protein gets dissociated and the resultant monomers are hydrated which was evident by fluorescence data and the difference spectral results with a 6nm red shift in the emission maximum and 2 nm blue shift in UV-absorption maximum arising out of perturbation of aromatic chromophores. The studies were performed both at native pH of 7.9 where the protein is in its oligomeric form and at pH of 10 where it is dissociated form and the results compared. The data showed that the solvent is excluded more from the protein vicinity in the dissociated state. PMID- 9753866 TI - Harmaline interactions with yeast invertase. AB - The effect of harmaline, a plant alkaloid has been studied on yeast invertase activity in the absence and presence of 50mM Na+ as a function of pH. Harmaline (1-3 mM) inhibited the invertase activity at pH 5.2, 6.8 and 8 both in the absence (44-92%) and (22-85%) of Na+ ions. Kinetic analysis revealed that harmaline is a non-competitive inhibitor of invertase, at pH 5.2 and 6.8 but at pH 8, it produced a mixed type of inhibition, Km increased by 450% and 175% and Vmax decreased by 82% and 63% in the absence and presence of 50mM Na ions respectively. The observed inhibition of invertase by harmaline was reversible in nature. These findings suggest that the presence of Na+ site is not a prerequisite for the inhibition of enzyme by harmaline. PMID- 9753867 TI - Prediction of structure of antenna proteins of photosystem II. AB - Membrane spanning regions of 43 kDa and 47 kDa antenna proteins of photosystem II of thylakoid membranes are theoretically predicted. Prediction of topology of chlorophyll-a and beta-carotene molecules in the proteins and interaction of the proteins with 33 kDa extrinsic protein on the lumenal side of thylakoid membrane is based on the findings reported earlier. Each antenna protein is predicted to have six transmembrane alpha-helices with twelve chlorophyll-a and five beta carotene molecules binding to it. Both N- and C- terminal ends are proposed to be on the stromal side of thylakoid membrane. The proposed structural model conforms to the reported experimental results from the literature. PMID- 9753868 TI - Poly-ADP-ribosylation of histone proteins of human kidney T1-cells in vitro following gamma-irradiation. AB - Poly-ADP-ribosylation of cellular proteins is involved with radiation induced damage and its repair. It has been observed that suspension of human kidney T1 cells in vitro attained elevated levels of poly-ADP-ribosylation due to experimental manipulations necessary for preparation of single cell suspension from monolayer cell cultures. These cells in suspension were exposed to various doses of gamma-rays with or without subsequent repair incubation. The PADPR of histones H3, H1 and H2B increased with increasing dose of radiation and decreased after 90 min or repair incubation. Concomitant with these changes, the affinity of histones to DNA in chromatin reduced immediately after irradiation. Normal affinity was reestablished after post-irradiation repair incubation. The results indicate that induction of poly-ADP-ribosylation of histone proteins by radiation and by manipulations to prepare single cell suspension involved different cellular components. PMID- 9753869 TI - Early and long-term effects of chronic low-dose radiation and coenzyme Q diet on the proliferation of rat spleen T cells. AB - Chronic whole-body irradiation (0.43 cGy/day; total doses 11, 23, and 35 cGy) caused non-monotonous long-term disturbances in rat splenic T lymphocytes proliferation. Immunosuppression observed after chronic exposure had no correlation with the splenic cell number and the decrease in the fluorescence intensity of Hoechst 33258-DNA complex in T lymphocytes. Small, but significant radioprotection was observed with Coenzyme Q diet immediately after irradiation. These results indicate that the changes in T cell immunity, T cell viability, and T cell DNA state after exposure to low dose radiation are not interrelated. PMID- 9753870 TI - Characterization of mouse fibroblast (NIH3T3) and fibrosarcoma cell lines (WEHI 164 and MFS 8) using PMRS. AB - Proton magnetic resonance Spectroscopy (PMRS) has been used to study the differences between immortalized fibroblasts and fibrosarcoma cells of different grade. One and two dimensional purged correlation spectroscopy (PCOSY) have been used to assess intact viable fibroblast and fibrosarcoma cells, and differences in the triglyceride, cellular metabolite, and cell surface fucosylation patterns between the three cell lines have been observed. The clinical implication of this study is the potential use of PMRS as an adjunct to conventional histopathology. PMID- 9753871 TI - Effect of alpha-tocopherol on mitochondrial electron transport in experimental myocardial infarction in rats. AB - The effect of alpha-tocopherol pretreatment (6 mg/100 g body wt/day, orally for a period of 90 days) on mitochondrial electron transport in myocardial infarction induced by isoproterenol (20 mg/100 g body wt, subcutaneously for two days) was studied in rats. A significant decrease was observed in the activities of isocitrate dehydrogenase, alpha-ketoglutarate dehydrogenase, succinate dehydrogenase, malate dehydrogenase, NADH dehydrogenase and cytochrome oxidase in heart mitochondria of isoproterenol administered rats. The cytochrome content and the oxidation of succinate in state 3 and state 4 decreased significantly in the cardiac mitochondria treatment. In alpha-tocopherol pretreated rats, the activities of TCA cycle enzymes, concentration of cytochromes and the oxidation of succinate in state 3 and state 4 were retained at near normal values, following isoproterenol administration. PMID- 9753872 TI - Determination of plasma oxalate with chloride ion insensitive oxalate oxidase. AB - A simple colorimetric method has been developed for determination of oxalate in plasma using a C1-insensitive oxalate oxidase purified from grain sorghum leaves. The ultrafiltered plasma collected in 3.5 N HC1 was pretreated with NaNO2 to avoid possible interference by ascorbate. The minimum detection limit of the method is 0.225 mg/l. The % recovery of the added oxalate was 91.5 +/- 5.0 (mean +/- S.D., n = 15). The coefficient of variation within and between batch were < 3 and < 5 respectively. The mean plasma oxalate concentration in healthy subjects was 0.29 mg/l. The method has the advantage over other enzymic methods that it doesn't require the removal of C1- prior to oxalate assay. PMID- 9753874 TI - Well-being and health. Background to the northern Finland 1966 birth cohort research. PMID- 9753875 TI - Changes and generations of the Finnish society in the 20th century. PMID- 9753873 TI - Verapamil and TTX inhibit +Vmax but differentially alter the duration of action potential of adult chicken ventricular myocardium. AB - Verapamil, a Ca2+ channel blocker, is also reported to block Na+ channels in mammalian heart and to modulate the repolarisation phase of cardiac action potential (AP). The Na+ channel blocking activity of verapamil and its implication to changes in repolarisation were studied on chicken ventricular strips where upstroke is due to highly TTX sensitive Na+ channels. At low doses verapamil (0.5-5 micro M) and TTX (0.1-0.5 nM) did not cause any significant effect on resting membrane potential (Em), maximal upstroke velocity (+Vmax) or AP duration (ADP). Higher concentrations of both verapamil (10-320 micro M) and TTX (1-40 nM) caused dose-dependent decrease in +Vmax and overshoot (Eov) without any change in Em. EC50 for the inhibitory effect of verapamil and TTX on +Vmax was 140 microM and 14 nM respectively. Na+ channels in adult chicken ventricular myocardium, therefore, seem to be more sensitive to TTX than their mammalian counterpart. Higher dosage of verapamil are needed to block Na+ channels in adult avian heart as reported for mammalian myocardium. Both verapamil and TTX caused dose-dependent changes in APD at-20 mV (ADP20) and at 90% repolarisation (APD90). TTX (1-40 nM) produced a decrease of 5-13% in APD20 and 4-12% in APD90 indicating a uniform hastening of the repolarisation process. Verapamil (10-320 micro M), however, induced 6-38% decrease in APD20 but 5-12% increase in APD90. Regression analysis of the relationship between changes in +Vmax and APD20 and APD90 in presence of TTX and verapamil exhibit significant linear correlation r for APD20 and APD90, being +0.965 for TTX and +0.978 and-0.898 for verapamil respectively. A linear correlation between inhibition of +Vmax and reduction in APD for TTX indicates the possibility that Na+ channel linked mechanism(s) underlie repolarisation process. Verapamil induced decrease in APD20 and increase in APD90 could be explained by the block of Na+/Ca2+ and K+ channels respectively. PMID- 9753877 TI - Obesity is a health problem. AB - Obesity is a very common and an increasing nutritional problem in Finland and in other western countries and also in circumpolar areas. Dietary factors like high intakes of fat and alcohol, and physical inactivity predispose to obesity. Childhood obesity increases the risk of obesity in adulthood. Obesity is associated with cardiovascular diseases, non-insulin-dependent diabetes mellitus and musculoskeletal disorders which cause work disability. Prevention of obesity should be more focused because treatment of obesity is difficult and the long term result generally poor. Multidisciplinary co-work is needed to find out how the factors of childhood are related to obesity in adulthood. PMID- 9753876 TI - Health trends in northern Finland. AB - Along with the process of the epidemiological transition, northern Finland has experienced an increase of cardiovascular diseases, some types of cancer, and accidental and suicidal deaths, particularly in the male population, while the females of northern Finland have shown rather favourable trends during the post war period. Northern Finland shows a clustering of severe health problems such as coronary heart disease, accidents and suicides in smaller areas, e.g. in north central Lapland, which records mortality rates 2-3 times higher than areas of lowest mortality. There is some indication of a high prevalence of smoking, increased serum lipids, blood pressure, body weight and alcohol consumption in the areas of highest morbidity and mortality, but evidence based on representative population samples is missing. Accidents and suicides are also common in the Sami (Lapp) area but coronary heart disease is rare, despite the unfavourable risk factor patterns. PMID- 9753878 TI - Antioxidants, infections and environmental factors in health and disease in northern Finland. AB - Recent studies have identified several factors which may affect human health and life expectancy in northern Finland. They have shown that antioxidants, infections, genetic or environmental factors may affect the development of and morbidity/mortality from cardiovascular diseases, cancer, diabetes mellitus and other diseases in the northern provinces of this country. Both the occurrence and mortality from coronary heart disease (CHD) is low in the northernmost part of the country, i.e. Mountain Lapland or the Saami area, compared with that in whole country or a neighbouring region to the south in central Lapland. The mortality from all diseases is also low in communities in Mountain Lapland, and high in central Lapland in communities such as Kittila and Kolari. High scrum antioxidants, alpha-tocopherol (vitamin E), albumin and selenium levels have been measured in men living in the northernmost part of the country, where the death rate from CHD is low. Low serum alpha-tocopherol and albumin levels were typical of men living in rural communities with high CHD mortality, e.g. Kittila community. Serum antioxidant levels were related to the diet; alpha-tocopherol increased with the consumption of reindeer meat and selenium with fish consumption. Our earlier studies have also identified a low Chlamydia pneumoniae IgA antibody titer in men living in Mountain Lapland compared with men in the neighboring region to the south in central Lapland with high CHD mortality. An elevated Chlamydia pneumoniae IgA antibody titer was associated with low serum alpha-tocopherol level. The people of Saami origin, an ethnic minority living in northernmost Finland, have a high apolipoprotein (apo) E e4 allele frequency and high serum cholesterol. They also have more apo A-IV-2 allele than most of the studied populations, and their HDL cholesterol levels are higher in apo A-IV-2/1 than in apo A-IV-1/1 phenotypes. Our earlier studies indicate that people living in northeastern Finland, west of smelters in Kola Peninsula may be exposed to heavy metals such as cadmium and mercury. Blood cadmium was related to blood pressure and high in men with arterial hypertensive disease. The investigations presented in this article form a good basis for further studies that clarify underlying reasons the health problems in the north. PMID- 9753879 TI - Intrauterine and life course factors in the aetiology of adult cardiovascular disease. AB - International comparisons of age-adjusted coronary heart disease mortality show the high rates in the eastern and northern Europe, and northern Finland belongs to the areas with the highest rates. This stresses the importance of further studies on the determinants of the disease in this area. There is growing evidence to suggest that patterns of early growth and other life course factors play an important role in the origins and development of cardiovascular disease (CVD), but understanding the processes which mediate these effects is limited. Low birth weight and some other birth measures are related to increased CVD mortality, hypertension and type 2 diabetes, even though inconsistencies between and within the studies exist. With the present available evidence there is a need to address the key issues of possible confounding of perinatal and early life measures with those in later life in relation to the CVD risk. There is a need to replicate studies and establish new ones by assembling cohorts where indicators of prenatal and postnatal growth have been previously recorded drawn from different populations living under different conditions. PMID- 9753880 TI - Trends and international comparisons in infertility in circumpolar areas. AB - Infertility is an increasing problem for both individuals and societies. The number of couples seeking treatment for infertility is increasing each year, and public interest seems to be rising along with the new treatment methods and the improving results. Male infertility is also of great interest now that several studies suggest a deterioration in the quality of semen in many countries, Finland being an exception. The assisted reproductive technologies have improved tremendously since the first child conceived by in vitro fertilization was born in 1978. The new techniques include e.g. intrauterine insemination (IUI), gamete intrafallopian transfer (GIFT), in vitro fertilization (IVF), intracellular sperm injection (ICSI) and various combination treatments. These treatments are costly and both physically and emotionally stressful, and the success rate varies according to the aetiology of infertility, the age of the woman treated and the method used. More information is needed about the aetiology and incidence of fertility disorders as well as about the availability of treatment in the circumpolar areas and the couples' opinions of treatment. Our own study population, which was drawn form the northern Finland birth cohort for 1966, provides an outstanding opportunity to study these issues, since data are available for the whole life course of the individuals, dating back to prenatal life. PMID- 9753881 TI - Polycystic ovary syndrome and hyperandrogenism as a risk factor for cardiovascular disease. AB - About 30% of infertility is caused by anovulation, associated most commonly with the polycystic ovary syndrome (PCOS). PCOS is a common endocrinopathy in women, especially at the age of 30 to 35 years, characterized by irregular menses, infertility and signs of hyperandrogenism. The pathogenetic mechanisms leading to PCOS are not fully understood, although several theories have been proposed. PCOS patients commonly have hyperinsulinemia and insulin resistance which are also known risk factors for the development of diabetes mellitus, hypertension and cardiovascular disease. However, it is not known, how well the presence of PCOS symptoms would predict the appearance of the long-term sequelae of insulin resistance. Also more data is needed e.g. of the role of intrauterine factors and birth weight in the development of PCOS and hyperandrogenism. PMID- 9753882 TI - To be born as a twin--risks and sequelae. AB - About 2.5% of infants born in Finland are twins. Twins have long been objects for genetic studies, which nowadays have large study groups, e.g. twin registers from a whole country. Twins can also be studied from another point of view: the special situation of being a twin, and its consequences on development and mental health. Perinatal mortality and morbidity are higher in twins than in singletons, and accordingly cumulative incidences of various handicaps are higher in twins. The human relationships of twins have their special features from the early beginning. Twins have to share the attention from the parents, and some parents resolve the situation by sharing the twins: "mother's child" and "father's child" may develop. Twins may be dependent on each other, and the inter-twin relationship can also be characterized by dominance-submissiveness. In adolescence, the time of getting independence from the parents, twins have also another task of development: they have to grow up from the co-twin dependence in order to become autonomous adults. In comparison to much bigger twin materials of genetic studies, follow-up studies in birth cohorts have their benefits, too. They give us a good opportunity to research the development of human relationships in twin families and their consequences on both somatic and mental health. PMID- 9753883 TI - Why do we need more information about the risk factors of the musculoskeletal pain disorders in childhood and adolescence? AB - Musculoskeletal pain disorders such as low back pain, neck pain and shoulder pain are a major and ever increasing public health problem among the working population in industrialized countries with social insurance. Especially the economic impact of these diseases on society has been rising, but the disorders do also produce a lot of pain and suffering to the people. It is an important challenge to the health care systems to prevent and treat these disorders, but at the moment poor understanding of the risk factors of these diseases has failed in giving any effective tools to control the musculoskeletal pain disorder epidemic. Most of the epidemiological studies made are cross-sectional and they do not extend to childhood and adolescence, when the organs are developing, achieving their loading strength and possibly being traumatized and starting their degenerative process. The longitudinal Northern Finland 1966 birth cohort study offers a unique opportunity to find early risk factors for musculoskeletal pain syndromes. PMID- 9753884 TI - Hearing and occupation. AB - Not only does the environment play a role as a source of risk factors for a hearing impairment, but a hearing impairment itself can adversely affect interaction with family members, workmates and friends, thus reducing social well being. The number of work-related hearing impairments has been decreasing for last five years, but noise-induced sensorineural hearing loss is still the second most common work-related disease in Finland. The financial burden related to occupational hearing impairments includes costs of compensation, salaries of screening personnel, equipment, maintenance costs, costs resulting from loss of work for the employer and referrals to specialist clinics etc, which until now have not been calculated in Finland. Numerous questions still remain to be answered regarding the association of age, socioacousis, occupation and leisure activities with the development of sensorineural hearing impairment. Can hearing impairment acquired in childhood or in early adolescence predict the development of occupational hearing loss? What is the interactive role of such factors as ageing, chemicals, diet, environmental noise, genetic susceptibility and the individual's other diseases in the development of noise-induced hearing impairment? PMID- 9753885 TI - Work ability of young adults. AB - This paper reviews the themes related to work ability of young adults. Premature working discapacity causes significant economical and social costs in Finland compared to other Nordic countries and has for that reason been studied most intensively there. Work ability is an interaction of social, environmental and individual factors such as physical fitness, coping skills, social support behaviour and health behaviour. Environmental factors influencing work ability can be concrete, like physical and chemical exposures or more abstract like unemployment. The work ability of ageing people has been studied intensively, while work ability of young adults has not been properly evaluated so far. The worsened economic situation in Finland has meant an excessive work load for those who still have work. At the same time the risk of permanent unemployment is increasing. This polarizationing affects especially the young because they may fail to enter the working life. Also factors related to childhood may affect and even determine work ability in adulthood. PMID- 9753886 TI - Risk factors and prognosis for psychiatric morbidity in children and adolescents. AB - Because of the new epidemiological studies during the past two decades our knowledge concerning child and adolescent psychiatric disorders has grown up. There exists data concerning the distribution of different disorders in the community. The development in the methodology of child psychiatric investigation has made it possible to study also the risk factors and prognostic features of child psychiatric disorders. In this paper risk and prognostic factors of various child psychiatric disorders are reviewed. All the findings from studies made in different countries are not suitable as such in Finland. No large scale epidemiological studies concerning the risk and prognostic factors of various child psychiatric disorders are available in Finland. The cohort-66 study offers a possibility to elucidate also these factors. PMID- 9753887 TI - Long-term outcome of migration in childhood and adolescence. AB - The effect of migration on the family and on the individual can be divided into three groups: cultural, changes in social environment and changes in the interpersonal relations. When successful adaptation is not achieved, acculturative stress may arise and somatic or mental disorders may develop. The finding of individual differences in people's responses to environmental conditions has led to search for vulnerability factors that increase people's susceptibility to stressors and for buffering influences that serve a protecting function under the same circumstances. The studies on migration should focus on somatic and mental health of the migrants, on achievements at school and at work, on protecting factors at different ages of migration and on the role of language acquisition and of social network on the adaptational process. The focus on this review is in childhood, adolescence and young adulthood. PMID- 9753888 TI - Education and mental disorders. AB - Mental health and education are crucial contributory factors in the welfare of the individual. Higher incidences of many mental disorders are found among populations with a low level of education. Difficulties in school and professional education may also predict adult mental disorders. The relationship between education and mental disorder has fairly seldom been analysed in empirical studies and further studies are therefore needed, as is an integration of educational policies in the prevention and care of mental disorders. In the Northern Finland Health and Well-being Study based on the Northern Finland 1966 Birth Cohort, education and its determinants is one of the main aspects studied. PMID- 9753889 TI - Psychiatric disorders and their predictors in young adulthood in Finland. AB - Mental disorders are one main focus of research interest in the 31 year study of the Northern Finland 1966 Birth Cohort Study. Mental disorders are quite common in young adulthood and they have a great impact on quality of life and working ability. Good national registers in Finland ensure the possibility to follow up treated incidence of severe mental disorders. On the other hand, a notable part of those who suffer from non-psychotic mental disorders do not receive any psychiatric treatment. That is why it is not possible to follow up psychiatric morbidity of the non-psychotic disorders from register data. In this review, principles of psychiatric diagnostics, known prevalences of psychiatric disorders in population and factors connected with mental disorder are briefly presented. Especially childhood predictors of mental disorders are reviewed. PMID- 9753890 TI - What kind of person is a thirty-year-old? AB - In this article we review the thirty-year-old person from the viewpoint of the development psychology, life span, physical health and family life. In his psychological development a person has at this stage become adult, but he still uses some psychic mechanisms and coping strategies which are typical for the adolescent. Intrapsychic world, the working role and the family life are stabilizing, but the changing over to the independence and working life can be more difficult today than earlier. Although with most people the adult life has settled down and physical and psychiatric illnesses are relatively rare, part of the thirty-year-olds are not independent, placed to the working life or are not healthy. In the following paper, main theories or findings in psychological and family life development and mental health of a thirty-year-old person are presented. PMID- 9753891 TI - Life control and health. AB - This paper describes the life control and its development, concentrating on relationship between life control and health. The main goal is to review the life control and health among Northern Finnish women and men. Life control and its connections with health and life situation make up the theoretical basis of the work. The definition of life control is based on Antonovsky's theory of a sense of coherence. The concept of life control here includes the following subconcepts: comprehensibility (how understandable are the internal and external environments), manageability (ability to influence one's life course at work and in one's whole life), meaningfulness (one's experiences of the meaning of the present or future events) and life satisfaction and human relationships. Future studies need to explain factors affecting the abilities of individuals to control their lives and reach sensible decisions regarding their health and lives overall. New ways of strengthening life control among those who have become marginalized in relation to society need to be identified. A model of factors relating to life control and relationships between such factors could be constructed. PMID- 9753892 TI - Updating of hygiene standards for carbon disulfide based on health risk assessment. AB - In 1995 the hygiene occupational standard values of carbon disulfide (CS2) were established in Poland: the maximum allowable concentration, eight-hour time weighted average (MAC-TWA)--18 mg/m3, and the short time exposure level (STEL)- 30 mg/m3. For lack of reliable retrospective data on the CS2 exposure levels in the work environment and the dose-response relationship, the following have been taken into account in establishing these values: the nervous and vascular systems are recognized as the main CS2 exposure targets; long-term exposure to CS2 in the work environment, exceeding 30 mg/m3, induces the toxic effect in the nervous and cardiovascular systems; chronic exposure to CS2 at concentration below 20 mg/m3 does not produce adverse effects in the peripheral nervous and vascular systems; coronary heart disease does not occur more frequently in workers exposed to CS2. Aiming at updating the 1995 MAC value for CS2 the authors carried out an analysis of the recent literature data on the relation between exposure levels and health risk. The results of clinical and epidemiological studies published in 1995-1997 did not provide evidence that adverse health effects in the cardiovascular and neurological systems in persons occupationally exposed to CS2 at concentration below 48 mg/m3 is likely to occur. The studies of the harmful effects of low CS2 concentration on the reproductive system have not proved that CS2 affects the embryo and fetus. Moreover, in Poland the employment of women under conditions of CS2 exposure (regardless of concentrations) during pregnancy and breast feeding is banned. Because the latest reliable studies have not indicated that chronic CS2 exposure at the level of 20-48 mg/m3 exerts toxic effect on humans, CS2 concentration of 18 mg/m3 as MAC-TWA and 30 mg/m3 as STEL, adopted in 1995, need not to be updated. PMID- 9753893 TI - Occupational profile and cardiac risks: mechanisms and implications for professional drivers. AB - Of the population of 900 drivers, 419 drivers with changes in the circulatory system were enrolled in the study, and the control group consisted of 150 healthy drivers. In both groups there were professional and non-professional drivers. Traffic accidents caused by these drivers were registered over a five-year period. Drivers with cardiovascular diseases and professional drivers were more frequently responsible for accidents than healthy and non-professional drivers. The comparison between the group studied and the controls indicated over a two fold increase in the number of accidents caused by sick drivers. PMID- 9753894 TI - The chemoprotective effect of coenzyme Q on lipids in the paint and lacquer industry workers. AB - The influence of coenzyme Q on the lipid parameters in the paint and lacquer industry workers is presented. The examinations were carried out in the group of 24 workers employed at the paint and lacquer production, who received coenzyme Q10 as a chemoprotective agent. Serum concentration of basic lipid parameters: total cholesterol (TC), high density lipoproteins (HDL), low density lipoproteins (LDL), triglycerides (TG); lipid peroxidation products: malonyldialdehyde (MDA) together with 4-hydroxynonenal (4-HNE) and two antioxidant enzymes: superoxide dismutase (SOD) and glutathione peroxidase (GPx) were examined. The above parameters were measured in workers exposed to organic solvents and then after 4 weeks of coenzyme Q treatment. In order to explain, whether the occupational exposure is responsible for the changed level of some parameters, the reference group, not employed in the paint and lacquer industry, was used. The results indicated that the preliminary blood serum concentration of MDA + 4-HNE in workers exposed to organic solvents was significantly elevated in comparison to the control group. Statistically significant decrease in MDA + 4HNE concentration was observed after coenzyme Q treatment what lead to the conclusion that coenzyme Q could be considered as a protective agent against lipid peroxidation in occupational exposure. The changes in other parameters were statistically insignificant. PMID- 9753895 TI - Calculation and comparison of average standardised mortality ratio in occupational cohort study. AB - The average standardised mortality ratios (SMRs) were calculated, by the methods of median, mean, Poisson and meta-analysis, across 85 occupational cohorts. The difference of average SMR between these methods was reduced with increasing number of cases. The SMRs from the Poisson and fixed/random effects models were close, however, the fixed and random effects models should be more appropriate as the Poisson model ignores variation from the characteristics of different cohorts. There were also wider confidence intervals for the random effects model than for the fixed effects model. Based on such calculations and comparisons of average SMRs, we would suggest that if a total cohort consists of several subcohorts, a summary SMR should be calculated by the fixed effects or random effects models, instead of the Poisson model. PMID- 9753896 TI - Environmental exposure to asbestos in asbestos cement workers: a case of additional exposure from indiscriminate use of industrial wastes. AB - The paper presents data on cancer risk, especially pleural mesothelioma and lung cancer, among the workers of asbestos cement plant who living in the vicinity of the plant, were also environmentally exposed to asbestos. In 1959 an asbestos cement factory was founded in the rural area of south-eastern Poland. Apart from chrysotile asbestos, crocidolite was used till 1985 chiefly for the manufacture of pressure pipes. The blue asbestos made up 15% of the mean annual tonnage of the processed asbestos. It was found that soon after asbestos production had started the process wastes were made available to local community, particularly to the workers of that factory. For over twenty years asbestos wastes of all kinds, both wet (process sludge) and dry (from pipe and sheet grinding) were exploited for the hardening of roads, paths, farmyards and sports fields and as construction material components. For the evaluation of cancer risk due to occupational exposure to asbestos a cohort of 1,526 workers employed in this factory was observed till the end of 1996. The cohort availability was 95.6%. Standardized mortality ratio (SMR) was calculated using the man-years method. The reference population was the general population of Poland. The results of the study demonstrated a statistically significant increase in the risk of a) pleural mesothelioma--over an 80-fold excess among males and over a 200-fold one among females; b) lung cancer in females--over a 6-fold excess; c) colon cancer in males--over a 3-fold excess. In the 1990 ten new cases of pleural mesothelioma in the cohort were reported. As compared to other asbestos-cement cohorts in Poland, observed at the same time, this cohort presented a very high risk of pleural mesothelioma. The analysis of 16 cases of pleural mesothelioma found in the cohort from 1987 to 1997 revealed 4 cases with very short employment period (3.5 months-5 years) including two cases with relatively short latency period (11-12 years). In order to find explanation of these findings, additional investigations were made. The epidemiological study indicated that all these persons were at the same time subject to non-occupational exposure associated with massive utilization of commonly available asbestos-cement wastes as road surface material. PMID- 9753897 TI - Sickness absence in a rubber plant in Poland. AB - The disease-related temporary incapacity for work, its causes and duration are essential factors in the assessment of health status of the occupationally active population. The aim of the present study was to investigate the main causes of work disability among the rubber industry workers, with special regard to sickness absence among workers directly enganged in manufacture. The study was performed in 1995 on a sample of 973 workers (456 males and 517 females) at a plant producing rubber footware. The number of days of work disability from a particular disease, frequency and duration per year were examined. The analysis concerned such parameters of sickness absence as the lost time rate, average duration of absence, and percentage of workers on a sick-leave. The results revealed that during the period under study the main medical causes of the sickness absence included: a) for males--cardiovascular diseases (48% of the total sickness absence), respiratory diseases (18%), gastrointestinal disease (8%) and the nervous system and sense organs diseases (8%); b) for females- cardiovascular diseases (24%), respiratory diseases (16%), pregnancy, childbirth and puerperium complications (11%) and neoplasms (10%). The sickness absence of workers directly involved in the manufacture appeared to be by 72% higher than that noted for workers of other departments, with the age- and gender standardized lost time rate of 4.74. The differences can be related mainly to a higher percentage of the sick in the group of 'production workers' (43%) as compared to the 'non-production' ones (28%). The findings of our study indicate that in the rubber industry workers a high rate of absence due to some groups of diseases may be associated with exposure to hazardous agents in their work environment. PMID- 9753898 TI - Environmental hazards for children in USA. AB - Children are not little adults. Their tissues and organs are rapidly growing, developing and differentiating. At various stages these growth processes create windows of great vulnerability to environmental toxicants. Children's patterns of consumption and exposure are very different from those of adults. Children's combination of disproportionately heavy exposure plus biologic vulnerability makes them very susceptible to injury caused by toxicants in the environment. Development and adoption of a child-centered agenda for research and risk assessment will be necessary if disease in children of toxic environmental origin is to be controlled, prevented and eventually eradicated. This agenda will need to be multidisciplinary. It should include epidemiology, pediatrics, exposure assessment, toxicology, and health economics. The ultimate goals of this agenda need to be (1) the identification of etiologic associations between environmental exposures and pediatric disease; (2) the elucidation of disease mechanisms; (3) improved treatment; and (4) prevention. PMID- 9753899 TI - Salud ocupacional. PMID- 9753900 TI - Knowledge coupling: implementing outcomes measurement through the disciplined and routine control of inputs. AB - The successful implementation of outcomes measurement is jeopardized by its application within systems that do not control inputs. This systemic flaw within health care can be corrected by placing critical knowledge that is now in the minds of individual practitioners into computer-based knowledge coupling tools that can (1) track and control the inputs to the system in a disciplined and routine manner and (2) process information without error or bias. Outcomes then become a natural by-product of what amounts to a paradigm shift in the conceptualization and implementation of health care. PMID- 9753901 TI - Outcomes measurement: application of performance standards and practice guidelines in managed behavioral healthcare. AB - Managed behavioral health care organizations (MBHOs) have increasing accountability to their customers for measuring and managing the quality of care and service rendered to consumers. Health plans, state and federal agencies, employers, and unions express continued and growing interest in MBHOs, utilizing evidence-based practice guidelines and incorporating performance standards into daily operations. Standards and guidelines are often built into the behavioral health care program design by the customer, established by accrediting organizations, developed and disseminated by research and academic institutions, or promoted by professional associations. PMID- 9753902 TI - Evaluating a state agency's case management services. AB - Lessons learned in developing and implementing a program evaluation model for a state case management program are described. Uniformity of standards, data collection, and uniform data formats are critical components to measuring system outcomes. PMID- 9753903 TI - Where does culture fit in outcomes management? AB - The authors describe the concept of cultural competence and ways in which culture, a structure of care variable, is important to the delivery of culturally competent care. The role of culture in outcomes assessment and management is explored. The culture of the patient, the health care professional, and the organization is examined as it influences the potential to deliver culturally competent care. Strategies for developing a culturally competent work force are proposed with examples from ongoing projects in a large metroplex in the southwestern part of the United States. PMID- 9753904 TI - Selecting outcomes tools in geropsychiatric home care. AB - Mental health services are rapidly shifting from acute-based care to community based care. More adults over the age of 65 years are being treated in the home or community for chronic mental illness. As care moves to the community, funding and regulatory sources are demanding a greater focus on measuring quality of care by assessing and evaluating outcomes. This article outlines the selection of outcome assessment instruments used in geropsychiatric home care. PMID- 9753905 TI - A survey of patient education postdischarge. AB - Through a telephone survey, the Discharge Call Service (DCS) was instituted with 206 patients discharged from two medical-surgical nursing units. The purposes of the survey were to (1) improve patient outcomes by assessing patient perceptions of their recuperation progress after discharge, (2) assess the patient's post discharge educational needs, (3) provide additional information concerning diagnosis, treatment, or medications requested by the patient, and (4) direct patients to the appropriate medical center or community resource as needed for further assistance or education. In general, patients believed their medical status was progressing as expected; nevertheless almost half of the patients surveyed needed additional information or specific directions concerning their self-care. Findings suggest the DCS is an effective strategy to enhance the patient's ability for self-care after discharge. The DCS is also an easily implemented and cost-effective enhancement of hospital patient education and health promotion activities. PMID- 9753906 TI - Children's use of summer camp health facilities: a longitudinal study. AB - This descriptive study explores the relationship between age, developmental level, and gender and school age campers' use of camp heath facilities. The sample consists of 370 campers (141 girls, 229 boys) who attended three summer camps between 1977 and 1990. Encounters recorded in the nurses' log books are analyzed. Accidents and injuries, communicable diseases, discomfort-related problems, and allergies are the most frequent reasons for visits. Gender has a significant effect on midseason and odd-time visits and visits for accidents/injuries and constitutional symptoms. Low users visited most frequently for accidents/injuries; and high users for viral syndromes. More boys were low users. Factors that influence children's visits to camp health centers need further study. PMID- 9753907 TI - Adolescents' perspectives of chronic illness: "it's hard". AB - An exploratory, qualitative study with tenets from grounded theory was used to elicit detailed descriptions of adolescents' chronic illness experiences. The philosophy of symbolic interactionism guided this study. Understanding the adolescents' experiences included exploring adolescents' perspectives of the following: (1) what it is like to have a chronic illness, and (2) how they deal with having a chronic illness. A sample of 23 adolescents, 13 to 16 years of age, and diagnosed with either diabetes, asthma, arthritis, Crohn's disease, or ulcerative colitis participated. Data collection involved the adolescents participating in open-ended interviews. The constant comparative method was used to analyze all data from the interviews. The findings revealed that having a chronic illness made life more difficult for the adolescents. Adolescents experience extra effort, restriction, pain, and additional worries because of having a chronic illness. However, adolescents also clearly had ways to help them deal with their illness. These strategies and recommendations for practice and future research are discussed. PMID- 9753908 TI - Self-management development in children and adolescents with diabetes: the role of maternal self-efficacy and conflict. AB - The overall purpose of this study was to investigate maternal self-efficacy and its relationship to maternal perception of the child's self-management of diabetes. The influence of conflict between mother and child was also examined. One hundred and four mothers of children, ages 8 to 17 years, who were attending summer diabetes camp, were asked to rate their own and their child's abilities to manage the child's diabetes. Overall, the mothers expressed a high degree of self efficacy in managing their child's diabetes and perceived their children as average or above in managing their own diabetes when compared with agemates with diabetes. Furthermore, mother's level of self-efficacy was significantly positively related to their perceptions of their child's self-management. In addition, almost one third (30%) of the mothers reported experiencing interpersonal conflict regarding how much responsibility the child should assume in managing their own diabetes. Mothers who rated their child's level of independence as low were three times more likely to report experiencing conflict. In the vast majority of cases, the child was the primary source of conflict. When hierarchical logistic regression was used to multivariately model children's independence, conflict with the child remained a significant predictor, above and beyond background, demographic, and important conceptual variables, including self-efficacy. PMID- 9753909 TI - Mothers' experiences caring for children with disabilities who require a gastrostomy tube. AB - In this qualitative study, mothers' experiences of feeding children with severe disabilities by a gastrostomy tube are described. Twelve mothers each participated in one, open-ended, home interview. Mothers gave detailed accounts of their activities and the tremendous stress involved in feeding the children. They described spending enormous time and energy seeking confirmation of the feeding problem and devising extraordinary practices to ensure the child's survival before "giving in" to the gastrostomy tube. Following gastrostomy tube insertion, they initially felt relief and disappointment, before customizing feeding and moving on. Mothers' suggestions for improving professional services are discussed along with implication for practice and research. PMID- 9753910 TI - An analysis of program and family costs of case managed care for technology dependent infants with bronchopulmonary dysplasia. AB - Caring for technology-dependent infants with bronchopulmonary dysplasia requires a wide range and intense level of services. Case management programs can offer comprehensive services to patients with complex needs. The Neonatal Pulmonary Program at Tulane University Medical Center is a case management program for infants with chronic pulmonary problems. The purpose of this study was to describe the costs of providing care for technology-dependent infants with bronchopulmonary dysplasia and to include the direct and indirect costs to families as well as the Program costs. The study population included 89 infants enrolled in the program from September 1987 through February 1992. Outpatient, inpatient, and professional staff costs were derived from hospital and clinic billing information; all other costs were determined through family interviews. Total costs for outpatient services were $59,627 (89), professional team members time $185,539 (89); inpatient services $1,144,930 (89), professional costs for inpatient services $88,946; direct health costs $32,543 (37) for home health care, equipment, medications, special diets; direct non-health costs $30,670 (37) for transportation, meals, child care, lodging during clinic visits and rehospitalizations and household expenses to accommodate equipment needs; and indirect costs $26,173 (37) for missed work days and employment changes. PMID- 9753911 TI - Endoscopic surgery in infants and children: new approaches for surgical problems. AB - Endoscopic surgery is rapidly becoming the most popular means of resolving surgical problems and is now performed with increased frequency in children. New technology has allowed for direct visualization of the surgical field using innovative surgical approaches and techniques. Decreased postoperative morbidity, shorter hospitalizations, and rapid recoveries are among the major advantages of this form of surgery. This article provides information concerning the benefits and potential risks of endoscopic surgery and reviews the major procedures that are currently available for infants and children. PMID- 9753912 TI - The importance of the nurse education act for pediatric nurses. PMID- 9753913 TI - Detection and removal of head lice with an electronic comb: zapping the louse! PMID- 9753914 TI - [The progress and methodologic problems of in vitro radioassay]. AB - The history of development of in vitro radioassays is reviewed. The basic principles and clinical uses of saturation analysis, competitive radioassay and immunoradiometric assay are described. The more rapid, sensitive, specific and accurate assays are required at all times. However, it is well known that autoantibodies or heterophilic antibodies in serum interfere with radioassay and cause false results. Here I present the technical problems of the assay in the presence of such interfering substances which need to be resolved and also discussed the discrepancy between bioactivity and immunoreactivity in the assay. PMID- 9753915 TI - [Clinical significance of 123I-BMIPP myocardial SPECT]. AB - 123I-BMIPP is a radioiodinated branched chain fatty acid analog, showing high accumulation as well as prolonged retention in the myocardium. Therefore, it is used as a metabolic imaging agent suitable for myocardial SPECT. After 123I-BMIPP is taken in the myocardium, it remains there mainly as a storage type fatty acid of triglyceride, and it shows different behavior by the stage of cardiac disease. Usually, we evaluate metabolic findings by early imaging (20 minutes) after intravenous injection of 123I-BMIPP, however, some times washout or fill in are seen in the case of myocardial infarction or hypertrophic cardiomyopathy by delayed imaging (after 4 hours). The mismatch of 201TlCl and 123I-BMIPP are useful for the diagnosis of stunned myocardium or evaluation of prognosis. However, behavior of 123I-BMIPP in myocardium is known still not completely. Therefore clinical significance of this examination is thought to be clarified by the stock of cases of various heart diseases. PMID- 9753916 TI - [A phantom study for the evaluation of the effect of the high uptake in the liver on technetium-99m myocardial perfusion SPECT images]. AB - The purpose of this study was to evaluate influences of the high hepatic uptake on parameters such as filtering, attenuation coefficient, and scatter correction at reconstructing of the myocardial SPECT images. Hepatic and cardiac spaces of a myocardial phantom (RH-2, Kyoto Kagaku), were filled with technetium-99m and a three-detector SPECT system (GCA 9300-DI, TOSHIBA) was used. The hepatic activity's influence was estimated from a qualitative percent regional scattering and the effects of attenuation and scatter correction were evaluated by a circumferential profile curve. Percent regional scattering increased in reverse to hepato-cardiac distance (HCD) and in proportion to hepatic to cardiac activity ratio (HCR). This tendency was observed the most significantly in the inferior region, followed by in the lateral, anterior and septal regions, declining in this order. An artifactual defect adjacent to the liver was observed when HCR is three and HCD is zero. However, when the Butterworth filter was used with small filtering-sizes and lower orders in combination with attenuation and scatter correction, the defects were decreased up to 15% at counts in the inferior region. This study showed that the hepatic to cardiac activity ratio, and the hepatocardiac distance should be considered for reconstruction of the SPECT images. PMID- 9753917 TI - [A modified method of cardiac functional analysis for ECG gated SPECT: study of functional G-maps]. AB - To evaluate the cardiac function accurately using ECG gated SPECT images, we performed a modified method of cardiac functional analysis (Functional G-maps). One hour after the intravenous injection of 1,110 MBq of 99mTc-tetrofosmin, gated SPECT data was acquired dividing a cardiac cycle into 12 frames. Every short-axis images were usually reconstructed using first 11 of 12 frames. The reconstruction of these images was repeated performing slice thickness correction. Because the apex-to-base length is different at any frame during a cardiac cycle, 10 slices of short-axis images were obtained with the same thickness for each frame. Subsequently each short-axis image was divided by 40 radii, and the time activity curve was generated from the total counts included in each segment plus both neighboring segments. Afterwards the curve fitting was performed using the second reverse Fourier function. From fitted curves and their differentials, we estimated a variety of parameters including Max (End-systolic count), Min (End diastolic count), %CI (Percent count increase), Uptake, PCR (Peak contraction rate), PDR (Peak distention rate) and CT (Contraction time). In 5 normal subjects, %Max was greater in the anterior and septal regions, whereas %Min was greater in the apex and lateral regions. %CI and %PCR were similarly greater in the septal, anterior and inferior regions. On the other hand, %PDR in the lateral or inferior region was lower than the values in the other regions. In conclusion, this modified method is expected to be useful for accurate assessment of regional cardiac function and myocardial perfusion. PMID- 9753918 TI - [Decision tree sensitivity analysis for cost-effectiveness of chest FDG-PET in patients with a pulmonary tumor (non-small cell carcinoma)]. AB - Decision tree analysis was used to assess cost-effectiveness of chest FDG-PET in patients with a pulmonary tumor (non-small cell carcinoma, < or = Stage IIIB), based on the data of the current decision tree. Decision tree models were constructed with two competing strategies (CT alone and CT plus chest FDG-PET) in 1,000 patient population with 71.4% prevalence. Baselines of FDG-PET sensitivity and specificity on detection of lung cancer and lymph node metastasis, and mortality and life expectancy were available from references. Chest CT plus chest FDG-PET strategy increased a total cost by 10.5% when a chest FDG-PET study costs 0.1 million yen, since it increased the number of mediastinoscopy and curative thoracotomy despite reducing the number of bronchofiberscopy to half. However, the strategy resulted in a remarkable increase by 115 patients with curable thoracotomy and decrease by 51 patients with non-curable thoracotomy. In addition, an average life expectancy increased by 0.607 year/patient, which means increase in medical cost is approximately 218,080 yen/year/patient when a chest FDG-PET study costs 0.1 million yen. In conclusion, chest CT plus chest FDG-PET strategy might not be cost-effective in Japan, but we are convinced that the strategy is useful in cost-benefit analysis. PMID- 9753919 TI - [Measurement of regional cerebral blood flow using one-point arterial blood sampling and microsphere model with 123I-IMP: correction of one-point arterial sampling count by whole brain count ratio]. AB - A correction method was proposed for measurement of the regional cerebral blood flow using one-point arterial blood sampling and a microsphere model with N isopropyl-p-(123I)iodoamphetamine (123I-IMP). Instead of using continuous arterial blood sampling octanol-extracted counts, one-point arterial sampling whole blood counts corrected by the whole brain count ratio using 1-minute planar SPECT images were employed. The experimental subjects were 189 patients with cerebrovascular disorders. 123I-IMP, 222 MBq, was administered by intravenous infusion. Continuous arterial blood sampling was carried out for 5 minutes, and arterial blood was also sampled once at 5 minutes after 123I-IMP administration. Then the whole blood count of the one-point arterial sampling was compared with the octanol-extracted count of the continuous arterial sampling. A positive correlation was found between the two values (r = 0.87, p < 0.001). The correction method was carried out by calculation as follows. The ratio of the continuous sampling octanol-extracted count (OC) to the one-point sampling whole blood count (TC5) was compared with the whole brain count ratio (5:29 ratio, Cn) using 1-minute planar SPECT images, centering on 5 and 29 minutes after 123I-IMP administration. Correlation was found between the two values (r = 0.72, p < 0.001). The following relationship was shown from the correlation equation. OC/TC5 = 0.390969 x Cn-0.08924. Based on this correlation equation, we calculated the theoretical continuous arterial sampling octanol-extracted count (COC). COC = TC5 x (0.390969 x Cn-0.08924). There was good correlation between the value calculated with this equation and the actually measured value (r = 0.94, p < 0.001). The coefficient improved to r = 0.94 from the r = 0.87 obtained before using the 5:29 ratio for correction. For 23 of these 189 cases, another one-point arterial sampling was carried out at 6, 7, 8, 9 and 10 minutes after the administration of 123I-IMP. The correlation coefficient was also improved for these other point samplings when this correction method using the 5:29 ratio was applied. It was concluded that it is possible to obtain highly accurate input functions, i.e., calculated continuous arterial sampling octanol-extracted counts, using one-point arterial sampling whole blood counts by performing correction using the 5:29 ratio. PMID- 9753920 TI - [Usefulness of 123I-MIBG scintigraphy for prediction of effect of beta-blocker therapy in dilated cardiomyopathy]. AB - To determine whether 123I-MIBG (MIBG) scintigraphy is useful for predicting the effect of beta-blocker therapy in patients with dilated cardiomyopathy (DCM), we studied MIBG scintigraphy in 11 controls and 9 patients with DCM before starting beta-blocker therapy. First, initial and delayed heart-to-mediastinum ratios (H/M ratio) of MIBG activity in patients with DCM were significantly lower than those in 11 controls, respectively (initial H/M; 1.8 +/- 0.3 vs. 2.1 +/- 0.3, p < 0.02, delayed H/M; 1.6 +/- 0.3 vs. 2.4 +/- 0.2, p < 0.0001), and MIBG washout rate from the heart was significantly higher in patients than in controls (washout rate; 33 +/- 7% vs. 22 +/- 4%, p < 0.0005). Second, beta-blocker therapy improved LVEF in 7 patients (improved group), while it resulted in deterioration of heart failure, followed by death in 2 patients (deteriorated group). Although initial and delayed H/M ratios in the improved group were not significantly different from those in the deteriorated group, respectively, MIBG washout rate was significantly higher in the deteriorated group than in the improved group (45 +/- 8% vs. 30 +/- 3%, p = 0.04). Our study suggests that DCM patients with markedly rapid MIBG clearance may be deteriorated by beta-blocker therapy. In contrast, there were no differences in LVEF and plasma norepinephrine between improved and deteriorated groups. In conclusion, 123I-MIBG scintigraphy is useful for predicting the effects of beta-blocker therapy in patients with DCM. PMID- 9753921 TI - [Assessment of enlarged left atrium with 99mTc-tetrofosmin SPECT and echocardiography]. AB - This study was performed to clarify the possibility of visualization and quantification with 99mTc-tetrofosmin (Tf) myocardial scintigraphy in cases with a large atrium demonstrated by trans-thoracic echocardiography (TTE). Myocardial SPECT was evaluated in 4 patients with mitral stenosis and 15 patients with mitral regurgitation. Left atrium was identified in 12 out of 19 cases from an antero-posterior projection. The Tf uptake ratio of the left atrium, which was defined as the ratio of ROI count of the left atrium divided by the ROI count of the left ventricle, showed a good correlation with the left atrial area obtained by both trans-thoracic and trans-esophageal echocardiography (r = 0.88 and 0.91, respectively), These data suggest that Tf myocardial SPECT is a useful method of evaluating left atrial enlargement. PMID- 9753922 TI - [A case of choroidal malignant melanoma in which 123I-IMP scintigraphy was useful for diagnosis]. AB - A case of choroidal malignant melanoma in which N-isopropyl-p-[123I] iodoamphetamine (123I-IMP) scintigraphy was useful for diagnosis is reported. A 62-year-old man first visited our hospital 3 years ago complaining of decreases in left eyesight. CT showed a tumor with an arcuate high attenuation area on the aural side of the optic disk in the left eye. A crescent high attenuation area, evidence of retinal detachment, was also observed on the nasal side of the optic disk. T1-weighted MR images showed low signal intensity in most of the tumor and a high signal intensity area was seen in the center, T2-weighted MR images showed homogeneous and marked low signal intensity area. Differentiation from a hematoma or a vascular tumor associated with bleeding was impossible based on CT and MRI. However, because late-phase images obtained on 123I-IMP scintigraphy showed marked high accumulation in an area corresponding to almost the entire left eye, left eye enucleation was undertaken under a diagnosis of malignant melanoma. Histopathologic examinations showed tumor growth mainly in the subretinal area. The melanin content of the tumor was high in the periphery and relatively low in the center. Infiltration was observed into the sclera and around the vortex vein outside the eyeball, but this change could not be detected by CT or MRI. 123I-IMP scintigraphy was useful not only for qualitative diagnosis of choroidal malignant melanoma, but for the determination of the extent of the lesion as well. PMID- 9753923 TI - [Reassessment of a combination of cerebrospinal fluid scintigraphy and nasal pledget counts in patients with suspected rhinorrhea]. AB - A combination study of cerebrospinal fluid scintigraphy and nasal pledget counts was performed using 37 MBq of 111In-DTPA in 12 patients with suspected rhinorrhea. A pledget was inserted and dwelled in each nasal cavity for 6 hours, with the patient prone during at least 30 minutes. A total of 18 studies was implemented and nasal pledget counting method successfully diagnosed all of CSF rhinorrhea. Diagnosis was possible when pledget counts were greater than 1 kcpm. In patients with persistent, intermittent and occult/no nasal discharge, rhinorrhea was found in 100% (5/5), 60% (3/5), 25% (2/8), respectively. Two cases only exhibited positive scintigraphy. MRI or CT cisternography should be first performed in patients with persistent discharge, but in patients with intermittent/occult discharge pledget counting method might take priority of other diagnostic modalities. In conclusion, nasal pledget counting method is a simple and useful tool for detecting rhinorrhea. PMID- 9753924 TI - [Use of 99mTc-labeled myocardial perfusion agents in the Kanto-Koshinetsu area]. AB - We sent out a questionnaire to investigate how the 99mTc-labeled myocardial perfusion agents are used in the Kanto-Koshinetsu area. Answers were obtained from 83 laboratories. Examinations using 99mTc-labeled agents were performed more frequently than those using 201T1 only in 10 of 83 laboratories. Main reasons for not using 99mTc-labeled agents were as follows: two injections required in the stress study, superiority of thallium-201 in the assessment of myocardial viability, a large amount of clinical data cumulated in thallium-201 imaging, and problems caused by abdominal uptake of 99mTc-labeled agents. Comparing purposes of the examinations using 201T1 and 99mTc-labeled agents, evaluation of myocardial viability was more common in 201Tl imaging, meanwhile, detection of myocardial ischemia was more common in imaging with 99mTc-labeled agents. It was pointed out that dual isotope imaging using 201Tl and 99mTc-labeled agents and simultaneous evaluation of myocardial perfusion and cardiac function are clinically feasible as new diagnostic procedures. PMID- 9753926 TI - [The surgical and endoscopic treatments of benign submucosal tumors of the esophagus, stomach and duodenum]. AB - Analysis of the results of various modes of treatment and follow-up of 160 patients with benign submucosal tumors (BST) was carried out. Most often histological type of BST was leiomyoma (43.7%). BST were mainly located in the stomach (68.7%). 22 patients underwent open operations, 37--endoscopical removal of BST, 101 patients were followed-up from 5 to 10 years. The analysis of the material makes it possible to come to the conclusions about advisability of endoscopical treatment. The results of which are not inferior to surgical method of treatment. In patients with absence or slow growth of BST endoscopic follow-up is valid. PMID- 9753925 TI - [20 years' experience of forced bouginage for cicatricial esophageal strictures]. AB - Method of orthograde and retrograde forced bouginage for treatment of 533 patients with cicatricial strictures of the esophagus was used during 1975-1995 years (486 procedures). In comparison with routine methods it has shown high effectiveness and safety. Perforation of the esophagus was detected only in 2 (0.43%) patients. Good and favourable immediate results were obtained in 95.1% patients. Follow-up results up to 20 years are available for 336 (71.8%) patients. Good result was obtained in 78.6% patients, favourable--in 16.1%, poor results--in 5.2%. It is emphasized, that the principal method for treatment of patients with cicatricial strictures of the esophagus is bouginage. The method of orthograde and retrograde bouginage of the esophagus contributes to steady remission of the disease in 95% of patients. PMID- 9753927 TI - [The diagnosis and treatment of neck wounds]. AB - The largest in Russia experience of diagnostics and treatment victims with side arms' wounds of the neck (492 patients) is represented. The authors subdivide allo wounds into superficial (170 cases without mortal outcomes) and profound ones (322 cases with total mortality rate as 6.8%). Classification of the injuries, their features, depending of the causes and the king of the weapon, are represented. Complex of diagnostic methods, which application depends on severity of the patients condition, is substantiated. The standard surgical approach in profound injuries of the neck is considered to be an anterior colotomy. PMID- 9753928 TI - [The surgical treatment of injuries to the abdominal aorta and inferior vena cava]. AB - The experience of treatment of 44 patients with traumatic and abdominal aorta and inferior cava vein is represented. All traumatic injuries of these vessels were accompanied by damages organs. Various kinds of vascular suture and angioplasty with synthetic prostheses were used. 17 operated patients died (38.6%). 19 patients were followed up for period from 1 to 8 years after surgery. Neither arterial nor venous blood flow disturbances were revealed. PMID- 9753929 TI - [The extent of bone resection in osteomyelitis of the chest wall]. AB - The experience in treatment of 40 patients with osteomyelitis and chondritis of the bones of thoracic wall is presented. 39 patients (of 40) underwent surgery with due regard to an adequate extent of resection within so-called borders of intact tissues. Critical analysis of literature and authors' own experience made it possible to establish strict borders of resection of damaged bones in this disease of thoracic wall frame and to obtain good results of combined treatment in patients over 50 years. Complete removal of chondral tissues and resection of bones within the area of probably normal anatomic formation of the bone, irrespective of the origin of osteomyelitis and chondritis (hematogenous, exogenous), have promoted recovery of the patients. There were no relapses during 1-7 year period. PMID- 9753930 TI - [The treatment of wounds of the anal canal and perineum]. AB - Analysis of processes of wound healing in 215 patients after operations for hemorrhoids, anal fissures and rectal fistulas was carried out. Clinico-cytologic control of healing of postoperation wound has shown that ultra-violet irradiation of wound surface is most effective at early stages of wound healing due to its pronounced bactericidal and antiinflammatory action. Low frequency laser irradiation intensifies tissue reparative and hastens healing of wounds. The proposed method for treatment of wounds by successive use of ultra-violet and laser irradiation of wound surface combined with conventional ointment medications, improves the results of treatment and decreases the rate of purulent complications 2 times, promotes rapid healing of wounds. PMID- 9753931 TI - [Wound treatment using the flow of an ozonized solution under high pressure]. AB - New method for treatment of infected and festered, among them wounds with lingering healing was developed. The method was based on local application of fine-dispersion ozonized solution stream high pressure. This stream of ozonized fluid is generated under pressure of 350 ATM with the help of "Device for hydro pressive treatment of wounds", ozonization of the solution is performed by the "OZh-2" apparatus developed by the authors. Application of the method of hydropressive ozonization provides fast cleansing of wound surface from pyonecrotic masses, promotes elimination of infection and thus substantially reduces the period of treatment of the patients. PMID- 9753932 TI - [The use of ultrasonic cavitation and specific application therapy in the combined treatment of suppurative wounds]. AB - Ultrasound treatment of the surface of festered wounds and application on them of specific heteroplasma were used in 46 patients for healing of festered wounds. The method provides broadening of the indication for application of primary suture on the festered wound and decrease of the duration of hospital stay of the patients 2.2 fold. PMID- 9753933 TI - [The arteriolymphatic administration of antibiotics in treating patients with suppurative-inflammatory diseases of the abdominal cavity organs]. AB - Experimental study of pharmacokinetics of Gentamicin was carried out in 23 dogs. Three different modes of its intralymphatic administration were used: intranodular inguinal, lymphotropic retroperineal and arterio-lymphatic. It has been established, that arterio-lymphatic mode of introduction of antibiotics (as well as regional endolymphatic and lymphotropic) warrants high concentration of antibiotics in venous blood and lymph. The peculiarity of arteriolymphatic administration is regional effect of accumulation of the antibiotic in organs and tissues of abdominal cavity of corresponding arterial region. Arteriolymphatic infusions of antibiotics have been introduced into complex of drug treatment of patients with pyogenic and inflammatory disorders of abdominal organs. The clinical testing has revealed high curative effect of this made of antibiotics administration. PMID- 9753934 TI - [Bronchiolo-alveolar cancer and its surgical treatment (a practical view)]. AB - The authors have followed up 243 patients with bronchioalveolar cancer of the lung from 1979 to 1995 years. The age of the patients varied from 39-83 years. The ratio men women was as 1.1:1.9, which distinctly differ from the indices of morbidity from lung cancer of the whole, 4 forms of bronchioalveolar cancer, according to conventional methods, were singled out. 198 patients died after operation. Favourable long term results were obtained only in nodular forms of bronchioloalveolar cancer. The main diagnostic methods were roentgenological examination tramstriracic puncture and repeated studies of the phlegm. PMID- 9753935 TI - [The treatment of acute thrombophlebitis in the varicose saphenous veins of the lower extremities]. AB - 682 patients underwent treatment for varicose veins of lower extremities (VVLE). 641 patients with thrombophlebitis of VVLE were operated: in 62.9% of patients urgent operation and in 37.1% delayed operation after conservative treatment were made. Operation procedure comprised removal of all thrombotic varicose veins and ligation of communicant veins. The treatment of acute thrombophlebitis affecting anatomically intact veins in 41 patients was conservative. Comparative evaluation of the results of treatment evidences, that active surgical policy is justified as it leads to a decrease of complication rate, shortening of hospital stay duration 2 times and reduction of disability period. Conservative treatment of acute thrombophlebitis in intact veins is advisable. PMID- 9753936 TI - [The treatment of acute endotoxicosis in diabetic patients with surgical diseases requiring emergency intervention]. AB - The aim of the study is the development of integral methods for early diagnosis and treatment of systemic pathobiochemical disturbances in patients with diabetes mellitus under conditions of acute surgical endotoxicosis as well as optimisation of surgical policy and drug pathogenetic correction. Blood lipids peroxidation (malonic dyaldehyde content in NADF and ascorbate-dependent systems, intermolecular sewings in erythrocytes membranes) and central hemodynamic parameters. The study was carried out in 199 patients with acute surgical diseases accompanied with diabetes mellitus. Peculiarities of the course of the disease were revealed in dependence of the form of blood lipoperoxidation. The scheme of intensive care in conditions of decompensation of diabetes mellitus aimed at correction of pathobiochemical disturbances in patients with acute surgical endotoxicosis has been developed. PMID- 9753937 TI - [The results of prostatic adenomectomy in patients with severe concomitant diseases]. AB - Immediate and long-term results of prostate adenomectomy were studied in 1549 patients, 322 of whom being of old age. In 1499 (96.8%) patients concomitant diseases were revealed: ischemic heart disease (934 patients), cardiosclerosis after 1-3 myocardial infarction (185), hemiparesis after acute cerebrovascular disturbances (74), diabetes mellitus (88), chronic lymphoid leukemia (5), cirrhosis of the liver (15), cancer (22) and true diverticula (15) of the urine bladder, drug-related polyallergy (16). 628 patients were radically operated in conditions of circulatory insufficiency of stage I-II. In 631 (40.7%) patients surgical intervention was carried out as urgent because of acute dysuria (hampering of urination) or to bleeding from tumor. Transvesical adenomectomy was carried out with hemostasis by 2 semipouch string removable sutures. In 89.5% of patients uncomplicated course of postoperative period was observed. Postoperative lethality in patients with concomitant diseases made up 3.2%. Causes of death were postinfarction cardiosclerosis (6.5%), after-effect of cerebrovascular stroke (5.4%), diabetes mellitus (5.7%), cirrhosis of the liver (6.7%). 6 months to 11 years after the operation 91.2% of the patients achieved good follow-up functional results of surgical treatment, in majority of the patients medical and social rehabilitation was observed. PMID- 9753938 TI - [The problem of iatrogenesis]. AB - A rise of iatrogenic injuries of late disturbing. It comprises the whole spectrum of negative psychological and physical impacts of the physician on the patient. The source of iatrogenia is violation of moral norms, insufficient professionalism, negligence and indifference. Organizational of independent medico-legal association is considered necessary which on the base of strict and efficient regulations will control correctness of the treatment, determine the extent of iatrogenic injuries and responsibility for each of them as well as ensure normal conditions for work of surgeons and their legal defence from an negative actions. PMID- 9753939 TI - [A device for the transcutaneous drainage of cavities]. PMID- 9753940 TI - [The arrest of an arrosive hemorrhage from the brachiocephalic trunk into the tracheobronchial tree resulting from a tracheal ulcer due to an endoprosthesis]. PMID- 9753941 TI - [The unsuccessful embolization of an aneurysm of the common hepatic artery]. PMID- 9753942 TI - [Multiple foreign bodies of the penis]. PMID- 9753943 TI - [Alternative approaches to the surgical treatment of complicated forms of colonic cancer]. PMID- 9753944 TI - Co-cultivation of conjunctival epithelial cells and Chlamydia trachomatis: electron microscopic findings. AB - This study used primary culture of rabbit conjunctival epithelial cells to investigate the infection process of chlamydia. The epithelial cells isolated from conjunctiva of rabbit were initially cultured for three weeks. After attaining confluence they were infected with Chlamydia trachomatis (C. trachomatis) serotype D, and after co-cultivation for 24, 48, and 96 hours, electron microscopic study was performed. An inclusion body, a characteristic finding of chlamydial infection, was observed in the vicinity of the nucleus after 24 hours of co-cultivation. It contained a large number of elementary and reticulate bodies and their intermediate forms. Infectious particles known as elementary bodies were noted in the inclusion as 20 to 30 microns sized round bodies with an electron dense core. Reticulate bodies were also noted; they too were round but somewhat pleomorphic and larger than elementary bodies. Some reticulate bodies multiplied actively by means of binary fission. In this study, we observed the characteristic changes of C. trachomatis-infected cells; this in vitro system might provide a suitable model for the study of some aspects of the pathogenesis of ocular chlamydia infection. PMID- 9753945 TI - Glutathione transferase (class pi) and tissue transglutaminase (Tgase C) expression in pterygia. AB - Pterygia are considered to be induced by predisposing factors such as the external toxic environment. Glutathione transferases (GSTs) have a role in the detoxication of toxic chemicals. Transglutaminases (TGases) are involved in apoptosis, cellular adhesion and the wound healing process. As their expressions may be changed in abnormal conditions, we evaluated the clinicopathological status of pterygia by immunohistochemical study with GST-pi and TGase C. Twenty one pterygia, two pseudopterygia and five normal conjunctival specimens were used. The formalin-fixed samples were embedded in paraffin blocks, which were subjected to be stained with anti-GST-pi and anti-TG polyclonal antibodies immunohistochemically. They were graded from negative to strong. Staining patterns of GST-pi ranged from negative to weak in normal conjunctival epithelium, while in pterygia and pseudopterygia, one was weak, seven mild, ten moderate and five strong. As for TGase C expression, normal tissues were weak to mild, but ten were mild, nine moderate and four strong in pterygia and pseudopterygia. The general staining patterns of GST-pi and TGase C were prominent, ranged from moderate to strong in pterygia and pseudopterygia with basophilic degeneration and keratinization. On the corneal side of pterygia, TGase C was strongly positive in the basal epithelium on destroyed Bowman's layer and in conjunctival fibrous tissue. We suggest that GST-pi and TGase C are responsible for the process of pathogenesis of pterygia and pseudopterygia. PMID- 9753946 TI - Immunopathogenesis of experimental melanin-protein induced uveitis. AB - The etiology of uveitis, a general term for inflammatory disorders of the uveal tract, has not been clarified. The purpose of this study is to investigate the immunopathogenesis during experimental uveitis induced by bovine melanin associated antigen (BMAA). Experimental melanin-induced uveitis (EMIU) was induced in male Lewis rats by injection of BMAA and delayed type hypersensitivity (DTH) testing was performed. Fluorescence-activated cell sorting (FACS) analysis was used to examine expression of cell adhesion molecules with antibodies to ICAM 1 and LFA-1. BMMA-induced uveitis resembled human AAU, with onset and peak at approximately 14 and 21 days after immunization, respectively. Signs of uveitis disappeared by 4 weeks postimmunization. Histologic study revealed major infiltration of the iris and ciliary body by numerous inflammatory cells and minor infiltration of the choroid. DTH testing showed that ears injected with antigen were more swollen than non-injected ears. FACS analysis demonstrated that ICAM-1 and LFA-1 expression increased during EMIU, with ICAM-1 expression higher than that of LFA-1. In conclusion, BMAA is uveitogenically active in Lewis rats. Immunopathogenesis appeared to be mediated by cell expressing ICAM-1 and LFA-1. PMID- 9753947 TI - NF-kappa B activation following optic nerve transection. AB - In order to elucidate in vivo neuronal cell death in the retina, and involvement of NF-kappa B in this process, we studied the degeneration of retinal ganglion cells (RGCs) and the activation of NF-kappa B after transection of the optic nerve of adult rat at 5 mm from the eyeball. The morphology of dying ganglion cells in the retinal ganglion cell layer was observed by light and electron microscopy, the activation of NF-kappa B was investigated immunohistochemically. Seven and 14 days post-axotomy, dying cells contained pyknotic nuclei. The death of retinal ganglion cells involved apoptosis, activation of NF-kappa B (p50 and p65) was prominent in a time dependent manner. We observed axotomy-induced NF kappa B activation, which may mediate apoptosis of retinal ganglion cells. PMID- 9753948 TI - Two-year follow-up of eyes without topical corticosteroid treatment after photorefractive keratectomy (PRK). AB - This study was performed to evaluate the result of photorefractive keratectomy (PRK) without topical corticosteroid treatment at postoperative two years. PRK was performed by Summit OmniMed excimer laser, using a 5.0 mm ablation zone in 51 eyes of 29 patients who were then followed up for more than 2 years. During this period, patients who showed myopic regression of less than 1.5 diopters(D) or corneal haze less than grade 2, were regarded as the favorable result group and those who showed myopic regression equal to or greater than 1.5 D or corneal haze greater than grade 1 were regarded as the unfavorable result group. Thirty-four of 51 eyes showed favorable results without any corticosteroid treatment, and 17 eyes showed unfavorable results. In this latter group preoperative mean refractive error (-7.94 +/- 1.58 D) was significantly higher than in the favorable result group (-5.14 +/- 1.30 D) (p < 0.01, t-test); there was, though no statistical difference in age, gender, or corneal thickness. The results were unfavorable in only two of 32 eyes suffering from moderate myopia (< or = -6.0 D), but in 15 of 19 showing high myopia (> -6.0 D). In eyes suffering from moderate myopia, routine topical corticosteroid treatment after PRK does not appear to be necessary. PMID- 9753949 TI - Comparative study of three phacotrabeculectomy procedures through a single incision. AB - The objective of this study is to evaluate and compare the effects of three combined phacoemulsification and trabeculectomy procedures procedures (phacotrabeculectomy) involving intraocular lens implantation through a single incision. Twenty-eight eyes of 28 patients suffering from chronic open angle glaucoma and chronic angle closure glaucoma were enrolled in this study. No stitch phacotrabeculectomy (Group A), modified T-flap phacotrabeculectomy (Group B) and phacotrabeculectomy with sutures (Group C) were performed in 11, eight, and nine patients respectively, who were followed up for 6 months. At the end of that period, the mean reduction of intraocular pressure was 6.39 mm Hg; in group A, B and C, the reduction was 3.27, 4.87 and 11.55 mm Hg, respectively. The procedure followed in group C was therefore most effective, and there was a statistically significant difference between the groups (p < 0.05). The survival rate of blebs was also marked in group C. There were no apparent differences in terms of visual improvement, complications and number of medications between the groups. This study suggests that the choice of procedure should be determined by the desired degree of pressure reduction. PMID- 9753950 TI - Peripapillary atrophy in normal and primary open-angle glaucoma. AB - This study was undertaken in order to determine the value of measuring peripapillary atrophy for the diagnosis and follow-up of patients with glaucoma, and to evaluate how closely peripapillary atrophy is related to structural and functional optic nerve damage in glaucoma. Magnification-corrected morphometry of photographs using a computer graphic program and automated static threshold perimetry were performed in 234 eyes of 141 patients with primary open-angle glaucoma and 139 eyes of 86 normal subjects. The groups were not significantly different in age, refractive error or disc area. Zones alpha and beta were significantly larger, total peripapillary atrophy was significantly more extensive, and zone beta occurred more often in the glaucoma group than in the normal group. The frequency of zone beta increased with advancing glaucoma stage. The areas of zones alpha and beta and total peripapillary atrophy increased significantly with decreasing rim/disc area ratio, rim area, and mean deviation, and with increasing vertical and horizontal cup-to-disc ratios and cup area. Correlation coefficients were generally higher for zone beta than for zone alpha. Peripapillary atrophy was greater in a sector in which the neuroretinal rim loss was more marked. These findings suggest that increases in the extent of peripapillary atrophy are related to the severity of glaucomatous optic nerve damage and visual field defects, and that peripapillary atrophy is useful for the diagnosis and progression of glaucomatous nerve damage. PMID- 9753951 TI - Retinal vessel diameter in normal and primary open-angle glaucoma. AB - The purpose of this study was to determine how closely peripapillary retinal vessel diameter is related to functional and structural optic nerve damage in primary open-angle glaucoma. Using optic disc photographs of 234 eyes of 141 patients with primary open-angle glaucoma and 139 eyes of 86 normal subjects, the diameters of the superior and inferior temporal retinal arteries and veins were measured at the optic disc border. On the basis of rim/disc area ratio, the glaucoma group was divided into four stages: early, more than 0.61; medium, 0.60 0.41; advanced, 0.40-0.21; far advanced, less than 0.20. In the normal group the diameter of the inferior temporal vein was the largest, followed by that of the superior temporal vein, the inferior temporal artery, and the superior temporal artery. The diameters of the inferior and superior temporal retinal artery were significantly smaller at the early and medium stage, respectively, whereas both inferior and superior temporal retinal vein diameters were significantly smaller at the far advanced stage. The diameters of the inferior and superior temporal retinal arteries correlated significantly with neuroretinal rim area (r > or = 0.48, P = 0.0001), mean deviation (r > or = 0.42, P = 0.0001), vertical cup-to disc ratio (r < or = -0.33, P = 0.0001), and peripapillary atrophy data (r < or = -0.14, P < 0.04). The results indicate that in primary open-angle glaucoma, vessel diameter becomes less as neuroretinal rim area decreases and visual field defects and peripapillary atrophy increase. Its evaluation can be helpful for the diagnosis of glaucoma and possibly also during follow-up. PMID- 9753952 TI - Anatomical and visual results of vitreous surgery for advanced retinopathy of prematurity. AB - We studied the visual acuity and anatomical stability of 101 eyes which had undergone vitreous surgery for advanced retinopathy of prematurity. All were followed up for at least six months. The anatomical and visual results of vitrectomy in 87 eyes were reviewed. The patients' average age at surgery was 9.6 months, and the average follow-up period was 37.6 months. To preserve the structure of the eyeball or to prevent further complications due to a shallow anterior chamber, a further 14 eyes underwent lensectomy. Total attachment was achieved in 20 eyes(23.0%), and partial attachment in 24(27.6%). Finally, 42 eyes(48.3%) were able to perceive light, while fixation and following occurred in 15(17.2%). Six eyes(6.9%) were able to identify form. The relatively useful vision achieved in cases of advanced retinopathy of prematurity in this study with long follow-up suggests that the rationale of vitreous surgery is to salvage functional retina and to prevent total blindness and other complications. PMID- 9753953 TI - The electroretinogram in chronic renal failure. AB - To evaluate functional changes of the retina in patients with chronic renal failure (CRF), we analyzed maximal combined response according to the recommendations of the International Society of Clinical Electrophysiology of Vision. Oscillatory potentials were extracted from maximal combined response by high pass filtering. Because most CRF patients suffer from hypertension, hypertensive patients were selected for the control group. Values recorded in CRF patients were compared with those recorded in hypertensive patients and in the normal control group. CRF patients underwent laboratory tests which included complete blood cell count and the determination of blood urea nitrogen, creatinine, natrium, and potassium levels. The parameters of electroretinograms obtained from CRF patients were compared with those obtained from the normal control group, in the former group all amplitudes were significantly lower and all implicit times except those of b-wave were significantly delayed (P < 0.05). In CRF patients, decreased b-amplitude of maximal combined response and delayed implicit time of oscillatory potentials 1, and 2 were significantly different from those in hypertensive patients (P < 0.05). CRF patients had anemia, and their blood urea nitrogen and creatinine levels appeared abnormal. There was, however, no clinical correlation between biochemical data and electroretinograms. Consequently, retinal function in CRF patients was severely damaged compared with control groups (i.e., normal and hypertensive patients). We suggest that these findings are result of anemia and uremia in addition to hypertensive retinal damage. PMID- 9753954 TI - Subconjunctival silicone oil drainage through the Molteno implant. AB - To describe silicone oil drainage from the vitreous cavity to the subconjunctival space through a Molteno implant. A 52-year-old aphakic man with a Molteno implant inserted after lensectomy, vitrectomy and intravitreal silicone oil injection visited our hospital. The Molteno tube was located in the anterior chamber with two silicone oil drops on it. Corneal dellen, induced by marked bleb-like elevation of the conjunctiva was noted during follow-up and excision biopsy was attempted. The conjunctival elevation consisted of innumerable microdrops of silicone oil enclosed by inflammatory subconjunctival tissue. Silicone oil as well as aqueous humor had drained through the Molteno implant and glaucoma implant surgery may thus not be appropriate for the control of intraocular pressure of aphakia by intravitreal silicone oil before removal of oil. PMID- 9753955 TI - [Effects of nitric oxide synthase inhibitor on skin blood flow responses to acute isovolemic hemodilution during halothane and isoflurane anesthesia]. AB - Eighteen dogs under halothane or isoflurane anesthesia were subjected for a study in skin blood flow of the forearm under acute isovolemic hemodilution. Hematocrit was intentionally reduced to about 50% of baseline in ten minutes and hematocrit changes were induced by exchange of blood with hydroxyethyl starch. The skin blood flow increase response, measured by laser Doppler flowmetry, to hemodilution was abolished after inhibition of endogenous nitric oxide synthesis by N omega-nitro-L-arginine methyl ester. Skin blood flow responses to hemodilution and N omega-nitro-L-arginine methyl ester were different between halothane group and isoflurane group. Nitric oxide may have an important part to play in vasodilation during acute isovolemic hemodilution, and it may be affected differently by halothane and isoflurane. PMID- 9753956 TI - [Hemorrhage exerts different effects on variabilities of heart rate and blood pressure in dogs]. AB - The aim of the present study was to evaluate the effects of hemorrhage on heart rate variability (HRV) and blood pressure variability (BPV) as indicators of autonomic nervous system (ANS) and hypovolemia. We induced hemorrhagic hypovolemia in 7 dogs by removing blood in graded stages (0%, 10%, 20%, 30%, 40% of the estimated blood volume; EBV). HR was unchanged during hemorrhage, while mean BP decreased significantly after 30% EBV hemorrhage. Low frequency component (LF: 0.04-0.15 Hz) of HRV significantly increased after 20% EBV hemorrhage but high frequency component (HF: 0.15-0.4 Hz) of HRV was not altered. LF of BPV increased significantly stepwise after 20% EBV hemorrhage and HF of BPV increased significantly after 30% EBV hemorrhage, showing that both LF and HF of BPV might indicate the degree of hypovolemia. During hemorrhage LF of HRV and BPV increased and HF of HRV was unchanged, indicating the shift of the autonomic balance toward sympathetic dominance. An excellent quantitative correlation between LF of BPV and the degree of hypovolemia was demonstrated during graded hemorrhage, while LF of HRV plateaued at its maximum value at 20% EBV hemorrhage. In conclusion, our study suggests that the spectral analysis of HRV and BPV during graded hemorrhage shows different characteristics in the quantitative evaluation of ANS and hypovolemia. PMID- 9753957 TI - [Does mixed venous desaturation during a bed bath indicate cardiopulmonary decompensation in postoperative cardiac patients?]. AB - The bed bath procedure consists of cleansing patients' body, passive position change, changing gown and making a bed. During the procedure, mixed venous desaturation was observed consistently in postoperative cardiac patients. We investigated the cause of the phenomenon in 22 patients undergoing cardiac surgery in their first postoperative day. The patients were breathing oxygen enriched air via a Venturi mask. Cardiac index (CI), transluminal SvO2, arterial blood gas, Hb, DO2, VO2, FIO2, A-aDO2 and Qp/Qs were measured before and during the bed bath, while the patients were in the supine and left lateral position, respectively. Mean 8.5 +/- 1.5 minutes were required to complete the bed bath. During the bed bath, SvO2 decreased from 71 +/- 7% to 59 +/- 9% (P < 0.001), and returned to the baseline 6.5 +/- 7.4 minutes after the completion of the bed bath. VO2 increased markedly from 128 +/- 27 to 194 +/- 47 ml.min-1.m-2 (P < 0.001), while DO2 increased slightly from 480 +/- 91 to 513 +/- 110 ml.min-1.m-2 (P < 0.05). Among the determinants of DO2, CI increased slightly from 3.3 +/- 0.6 to 3.6 +/- 0.8 l.min-1.m-2, Hb remained unchanged and SaO2 decreased from 98.5 +/ 0.8 to 98.0 +/- 1.1%. FIO2 also decreased, while A-aDO2 and Qp/Qs remained unchanged. There was a negative correlation between VO2 change and SvO2 change, but no correlation between DO2 change and SvO2 change. There was a positive correlation between SaO2 change and SvO2 change, as well as between FIO2 change and SaO2 change. Therefore, the major cause of mixed venous desaturation was not the decreased DO2 or cardiopulmonary decompensation but the increased VO2 due to increased activity of the skeletal muscles. However, the decrease in SaO2 due to markedly increased O2 demand and the limited increase in CI might partially contribute to the marked decline in SvO2 through the limited increase in DO2. PMID- 9753958 TI - [Effect of epidural anesthesia on airway constriction induced with methacholine or capsaicin in cats]. AB - The choice of epidural anesthesia for patients with bronchial asthma is controversial. We studied the effect of epidural anesthesia on airway constriction induced by methacholine or capsaicin in cats. Cats were anesthetized with pentobarbital and mechanically ventilated. Peak airway pressure and compliance, as well as cardiac sympathetic and vagal nerve activity were recorded. We sprayed 0.2% methacholine of 0.2% capsaicin into the trachea to produce airway constriction, and 15 min after drug spray we injected 2% lidocaine 1.0 ml into the epidural space. Methacholine increased peak airway pressure by 25% and decreased compliance by 26%. Capsaicin increased peak airway pressure 20% and decreased compliance 22%. After epidural anesthesia, cardiac sympathetic nerve activity decreased to 40% and 44%, vagal nerve activity decreased to 92% and 61% of control values in methacholine and capsaicin groups, respectively. However, here were no changes in the peak airway pressure and compliance in the two groups. These results suggest that epidural anesthesia, even if epidural anesthesia decrease sympathetic nerve activity, has no effect on the airway constriction induced with methacholine or capsaicin. PMID- 9753959 TI - [Effects of anesthetics on somatosensory evoked potentials by median nerve stimulation in human--analyses after single administration in volunteers]. AB - In the present study, effects of midazolam, thiopental sodium, propofol, and nitrous oxide upon SEP in a clinically used dose were investigated on 24 male volunteers. In addition, antagonistic actions of flumazenil and naloxone against effects of midazolam and nitrous oxide, respectively, on SEP were studied. Midazolam had no effect on latencies of N 20 and P 25, but increased latency of P 45 and attenuated P 100 amplitude. Flumazenil reversed these effects of midazolam of P 45 latency and P 100 amplitude to their control values. While thiopental sodium and propofol suppressed P 100 amplitude, they had no effect on N 20, P 25, P 45 latencies. Nitrous oxide elongated latencies of N 20, P 25, P 45 and decreased P 100 amplitude. Naloxone reversed the effects of nitrous oxide on N 20 and P 25 latencies without affecting increased P 45 latency and attenuated P 100 amplitude. These results suggest that midazolam might have an analgesic action of suppressing cortical sensory neurons, whereas thiopental sodium and propofol have no effect on neurons in the primary sensory cortex. The finding that naloxone antagonized the increased latencies of N 20 and P 25 by nitrous oxide could be explained by the analgesic action of nitrous oxide that could be mediated by opioid receptors. The results also indicate that electrical activities of the cortical neurons in the associated area are more susceptible to psychotropic agents than those in the primary sensory cortex. The effects of anesthetics on SEP appear to reflect their characteristics of functioning mechanisms on cortical neurons. Analysis of SEP is, therefore, useful for the assessment of the mechanism and the acting site of anesthetics in the sensory cortex. PMID- 9753960 TI - [Two cases of circulatory failure after local infiltration of epinephrine during tonsillectomy]. AB - We experienced two cases of circulatory failure after local infiltration of 0.0005% epinephrine solution for the purpose of prophylactic hemostasis during tonsillectomy under sevoflurane anesthesia. Case 1: A 14 year-old girl developed ventricular bigeminy, tachycardia and hypertension following infiltration of the epinephrine solution 6ml around the tonsil. Sinus rhythm returned with intravenous lidocaine 40 mg and propranolol 0.4 mg. However, the patient showed gradually decreasing heart rate, depressed ST segments and inverted T waves and poor peripheral circulation. Her blood pressure decreased abruptly at the same time and finally the pulsation of the radial and femoral arteries was not palpable. She was treated with intravenous ephedrine in vain. Therefore, she received intravenous epinephrine and cardiac massage, and then recovered from the circulatory failure with her ECG showing normal sinus rhythms. Emergence from the anesthesia was smooth. Her cardiac failure may have been caused by the decreasing cardiac contraction and the increasing afterload due to the vasoconstriction after the intravenous beta-blocker. Case 2: An eleven year-old boy showed ventricular tachycardia and hypertension after infiltration of the epinephrine solution 11.5 ml around the tonsil. Lidocaine was given intravenously. This restored sinus rhythm but the ST segments on his ECG were elevated. ST segments became normalized after intravenous nitroglycerin. However, pulmonary edema developed suddenly, and it was cured by intensive treatment. His ventricular tachycardia and hypertension after the local administration of epinephrine were presumably responsible for the acute heart failure causing the pulmonary edema. Our experience suggests that the maintenance of cardiac function and the reduction of afterload are important to overcome the circulatory disaster following the local infiltration of epinephrine. PMID- 9753961 TI - [Reduction of pain on injection of propofol: a comparison of fentanyl with lidocaine]. AB - We compared the effect of fentanyl and lidocaine on pain during injection of propofol. One hundred and sixty patients premedicated with midazolam were randomly allocated to one of four groups (n = 40, respectively); Group C, propofol 2 mg.kg-1; Group F, fentanyl 0.1 mg 3 min prior to propofol; Group L, lidocaine 40 mg added to 200 mg propofol; Group FL, fentanyl 0.1 mg 3 min prior to propofol mixed with lidocaine 40 mg. Propofol was injected via a vein on the dorsum of the hand in a half of the patients in each Group or the forearm vain in the other half. The incidence of pain was significantly less in both Group F (40%) and Group L (35%) compared with Group C (80%, P < 0.01). There was no significant difference in the incidence of pain between Group F and Group L. Group FL had the least incidence of pain (5%) of all Groups. Injection via the forearm vain tended to reduce the severity of pain compared with the vain on the dorsum of the hand. The time until loss of consciousness was significantly less in the groups receiving fentanyl than the groups without fentanyl (P < 0.01). In conclusion, prior administration of fentanyl is as effective as premixing of lidocaine in preventing the pain on injection of propofol, and the simultaneous application of them may abolish the pain. PMID- 9753962 TI - [The duration of action of vecuronium is unchanged when administered with propofol]. AB - We examined the difference in the duration of action of vecuronium injected through a venous line infused with propofol and the duration of vecuronium infused into another venous line without propofol, in order to investigate the interaction between vecuronium and propofol within the intravenous lines. The subjects of the study are 8 patients, (ASA grade 1 or 2, aged from 20 to 50 years, 6 males and 2 females), who had undergone elective operations under general anesthesia. The mean duration of action of vecuronium injected through the venous line infused with propofol was 32.3 +/- 9.0 (min), while that for vecuronium injected through the venous line without propofol was 32.1 +/- 8.6 (min). There was no significant difference in the duration of action of vecuronium between the two conditions. We conclude that since vecuronium can be injected through the venous lines infused with propofol without interaction, there is no need for an additional venous line without propofol when vecuronium is administered under propofol anesthesia. PMID- 9753964 TI - [Anesthetic management of a patient with a right kidney tumor associated with complete occlusion of the inferior vena cava by tumor embolism]. AB - We gave anesthesia to a patient with a right kidney tumor associated with complete occlusion of the inferior vena cava (IVC) by tumor embolism. The upper end of the tumor embolism was below the junction of the IVC and the hepatic vein, and the operation was considered possible by simply clamping the IVC. To prevent complications including pulmonary embolism, the circulatory change at the time of clamping of the IVC, and massive bleeding, monitoring was made by pulmonary artery catheter and transesophageal echocardiography, and extracorporeal circulation was prepared. The blood pressure was stable and massive bleeding did not occur at the time of clamping of the IVC, because the IVC was completely occluded. The monitor showed no signs of pulmonary embolism. In a case of kidney tumor with tumor embolism in the IVC, it is necessary to be fully prepared for pulmonary embolism, the change of blood pressure before and after clamping of the IVC and for the bleeding at the time of IVC excision. PMID- 9753963 TI - [Anesthetic management for Fontan procedure without the use of cardiopulmonary bypass]. AB - We examined the anesthetic management for Fontan procedure performed without the use of cardiopulmonary bypass (Group N, n = 7) and that for equivalent procedure under cardiopulmonary bypass (Group E, n = 10) retrospectively. In Group N, surgical repairs of major vascular system were performed while bypassing the superior or inferior vena cava to the right atrium. The use of anesthetics and vasoactive agents was similar in both groups. Patients in Group N had significantly less blood loss and were extubated significantly earlier than those in Group E. However, significant metabolic acidosis was noted in Group N when reconstruction of the vascular system was completed and so-called Fontan circulation was initiated. Fontan procedure without the use of cardiopulmonary bypass may have advantage of less impairment for the cardiac performance and the pulmonary vasculature. However, its anesthetic management is another challenge to the anesthesiologist and requires meticulous control of both optimum preload and vascular resistance of the pulmonary artery. PMID- 9753965 TI - [Anesthesia and perioperative management in infants with Chiari type II malformation]. AB - From July 1991 to June 1997. 15 neonates with Chiari type II malformation were treated at our institution. Four of them required posterior fossa decompression and cervical laminectomy for hindbrain decompression. We report anesthesia and postoperative management in these four patients. They had a fetal diagnosis of hydrocephalus and was delivered by caesarean section. They underwent Ommaya reservoir placement for drainage and repair of myelomeningocele in the neonatal period. They developed respiratory depression as apneic spells or retraction with or without swallowing difficulties and underwent posterior fossa decompression and cervical laminectomy at 20-87 days of life. One patient died of asthma at the age of 2 years and 8 days and others are doing well. Patients with this malformation may develop respiratory depression such as apneic spells and vocal cord paralysis even if the intracranial pressure is well controlled and they should be monitored carefully for the signs of apnea and the compromised airway. PMID- 9753966 TI - [The relationship between respiratory function and the change in end-tidal nitrous oxide concentration]. AB - The aim of the present study is to clarify the relationship between respiratory function and the rate of change in alveolar anesthetic concentration. We measured the concentration of end-tidal nitrous oxide (N2O) when 50% N2O was administered to 15 patients of ASA I possessing normal respiratory function during the course of propofol-100% oxygen anesthesia. All patients were ventilated at a rate of 8 10 ml.kg-1 x 8 times per minute using a conventional anesthetic ventilator with semi-closed circuit and 4 l.min-1 inflow of fresh gas. Arterial CO2 partial pressure was maintained at 36.2 +/- 1.8 mmHg and no significant circulatory change was observed while N2O was administered. The rate of increase of end-tidal N2O concentration in poor FEV1.0/FVC% group was significantly slower than that in high FEV1.0/FVC% group, while there was no relation between %VC and the end-tidal N2O concentration change. Since N2O is an inhaled anesthetic, it is well considered that the effect of FEV1.0/FVC% may be observed in other inhaled anesthetic although the magnitude of the effect may vary. The present result suggests that respiratory function, especially FEV1.0/FVC%, is an important factor affecting the rate of change in alveolar anesthetic concentration and, in lower FEV1.0/FVC% group, it takes more time to achieve the intended alveolar concentration. PMID- 9753967 TI - [Anesthesia for a patient with von Willebrand disease and asthma]. AB - An emergency appendectomy was performed in a patient with von Willebrand disease (vWD) and asthma. vWD is a hemostatic disorder in which the patient lacks von Willebrand factor, playing a critical role in the mediation of platelet adhesion. A 12-year-old boy with vWD underwent an appendectomy under general anesthesia. The patient had not previously received any medication for the treatment of vWD. His bleeding time was over 18 minutes. Confact F injection improved it to 6 minutes. The patient was given Confact F during the perioperative period and no abnormal hemorrhage occurred. Heat-treated factor VIII concentrates can compensate for the low concentration of von Willebrand factor and they have been designed by several different companies and individual researchers with slight variations in composition among the various products. This indicates that some of these products can not be effective in compensating for vWD. In this particular case, Confact F was suitable for this patient with vWD (II A type). In addition, the patient was suffering from asthma. To avoid introducing any superfluous irritants that might induce an asthma attack, we removed the endotracheal tube under deep anesthesia and maintained the airway with a mask until the patient became conscious. PMID- 9753968 TI - [Pulmonary artery injury caused by mal-manipulation of a pulmonary artery catheter]. AB - A 71-year-old male was scheduled for aortic valve replacement. After tracheal intubation under high-dose fentanyl anesthesia, a pulmonary artery balloon catheter was inserted via the right internal jugular vein. However, after withdrawing the catheter with the balloon inflated, we found abrupt massive bleeding from the endotracheal tube. Fortunately, hemorrhage stopped with PEEP (20-25 cmH2O). Bronchofiberscopy could not detect the bleeding point. Chest X-ray showed the pulmonary artery balloon catheter in the right pulmonary artery and atelectasis of the middle and lower lobes. Five days later, he was extubated without rehemorrhage. Three weeks after hemorrhage, the patient successfully underwent an aortic valve replacement, and was discharged on the 25th post operative day. PMID- 9753969 TI - [A comparison of the grade of laryngeal visualisation;--the McCoy compared with the Macintosh and the Miller blade in adults]. AB - Effectiveness in visualization of the vocal cord during orotracheal intubation with McCoy (McC) compared with Macintosh (Min) and Miller (Mil) blades were investigated. After an institutional review board approval, 117 patients for elective surgery under general anesthesia requiring tracheal intubation were investigated. Five board certified anesthesiologists tried to visualize the vocal cord of a patient three times with the three different types of laryngoscope. Total of 351 intubation attempts were studied. The view obtained at laryngoscopy with each of the three blades was recorded as follows. Grade 1. If most of the glottis is visible. Grade 2. If only the posterior extremity of the glottis is visible. Grade 3. If no part of the glottis can be seen. Grade 4. If not even the epiglottis can be exposed. Eight-two Grade 1 views were obtained with McC, 72 with Mil and 47 with Min, respectively. Thirty-three Grade 2 views were obtained with McC, 36 with Min and 24 with Mil. Two Grade 3 views with McC, 34 with Min and 14 with Mil were obtained. Seven Grade 4 views were obtained with Mil. The grades of laryngeal visualization with McC were significantly lower than those with Min and Mil. PMID- 9753970 TI - [The evaluation of the pre-operative interviews using information sheets]. AB - This investigation deals with patients of more than 15 years of age and family members of children younger than 12 years of age to evaluate the pre-operative interviews using information sheets. The information sheets describe the anesthetic management and complications in a simple style. Sixty% of the patients and 75% of the children's family felt anxiety about the anesthesia and/or the operation (P < 0.05). More than a half of the patients did not want to receive informations about the anesthetic management and the risk of anesthesia. On the other hand, 9% of children's family did not want to know informations about the risk (P < 0.05). More than 80% of patients read the information sheets after the pre-operative interviews and about a half of patients answered that their anxiety before the surgery decreased. In this investigation, the children's family wanted to have information about the anesthesia or the operation more than patients themselves. The pre-anesthetic interviews using information sheets is useful to give information about anesthesia and to relieve anxiety of the patients and the children's family. PMID- 9753971 TI - [Accuracy of measuring nitric oxide by the chemiluminescence: effect of measuring chamber pressure]. AB - There are two ways of measuring nitric oxide (NO) by chemiluminescence; one reduces pressure in the NO reaction chamber, and the other does not. Devices with a reduced pressure chamber tend to be more accurate, but can be bulky and cumbersome to use. Devices without reduced pressure chamber can be useful because of their smaller size and short warm-up time. We compared the accuracy of two methods by measuring delivered gas concentrations after changing ventilatory parameters. The values of nitric dioxide (NO2) measured varied more widely than those of NO. NO2 measurements were particularly variable with changes in humidity. We consider the accuracy of NO measured by the chemiluminescence devices to be accurate for clinical monitor, but the accuracy of NO2 measurement must be improved to make NO therapy safer. PMID- 9753972 TI - [Analysis of fusion waves created with temporal pacing]. AB - Fifty-five beats of fusion waves were recorded continuously with an audio digital tape and the tape was re-played for analysis. A 45-year-old male (56 kg, 175 cm) with cervical spondylosis was scheduled to undergo laminoplasty of the cervical vertebral (C2-C6). A temporal ventricular (VVI mode) pacing lead was inserted from the right cubital vein to the right ventricular apex for preventing bradycardia while manipulating the medulla. The height of the R wave decreased gradually and the depth of S wave increased in the earlier period of fusion beats and it was reversed later. The narrow QRS width indicated that the electrode was placed near the cardiac conducting system. The gradually increasing intervals between P waves activated the pacing, and the P wave intervals recovered inhibiting the pacing. During the recovery phase, some beats were still activated by pacing instead of depressing the rate below the original rate. These beats suggest the importance of considering the atrial-ventricular conducting time. Arterial pressure fluctuated only slightly during the 'fusion beats, suggesting that despite the abnormality in the cardiac conduction system due to pacing, contraction of the ventricular muscles was only slightly affected in this case. PMID- 9753973 TI - Internet survey research and consent. PMID- 9753974 TI - InfoRetriever: rapid access to evidence-based information on a hand-held computer. PMID- 9753975 TI - Modeling and evaluation of continuity of care in a staff model HMO. AB - The concept of continuity in medicine refers to the delivery of care in an uninterrupted and coordinated manner and in accordance with the patient's medical needs. Many diseases and symptoms require serial observation and treatment over long periods and are interwoven with the patient's personal and social circumstances. We believe that the depth of a primary care provider's understanding of a patient is directly proportional to the total length of interaction between the patient and provider. We have reviewed the published studies in this area, and modeled continuity of care in an HMO clinic. The patients' ages, their visiting patterns, and the length of their interactions with their doctors were synthesized stochastically. We now propose a new way of defining continuity of care, the Fundamental Continuity of Care Index (FCCI), and recommend its use. Hospitals, insurance companies, and governments should be aware of the benefits of continuity and all physicians should commit themselves to longitudinal care for their patients. PMID- 9753976 TI - Computer-supported identification and intervention for diabetic patients at risk for amputation. AB - We used the fully automated medical record system at Kaiser Permanente of Ohio to direct appropriate interventions to diabetic patients at risk for amputation. The computer identified all patients with a diagnosis of diabetes, reminded physicians, at the moment of care, of the need to enter the patient's risk status for amputation, and kept track of patients at medium or high risk for amputation who were due for an evaluation with education in the podiatry department. Two years and four months after activation of this reminder system, the risk level had been determined for 76% of the diabetic population (n = 10,000), and two thirds of those at medium or high risk had received the appropriate intervention. In patients in the medium and high risk groups, the risk ratio for amputation was 17.5. PMID- 9753977 TI - Improving the human-computer interface: a human factors engineering approach. AB - Human factors engineering involves the application of information about human behavior and characteristics in the design and testing of products, systems, and environments. A computing system's interface is developed on the basis of potential users' capabilities and limitations, the users' tasks, and the environment in which those tasks are performed. When human factors engineers work with users, subject-matter experts, and developers to design and test a system, they analyze and document users' tasks and requirements and develop prototype designs. Usability studies are conducted and user errors are analyzed to identify problems and develop recommendations for improving the human-computer interface. PMID- 9753978 TI - Electronic medical records for prenatal patients: challenges and solutions. AB - A longstanding impediment to successful medical computing is resistance on the part of physicians. Interaction with many medical computing systems is difficult, requiring the physician to spend valuable time and energy trying to figure out how to get the machine to do what needs to be done. In developing encounter forms for use in prenatal medical records, we confronted the challenges involved in designing a computing system that provides an intuitive and physician-friendly method of recording clinical data. In trying to meet those challenges, we also learned about how to evaluate a medical computing system for flexibility and ease of use. PMID- 9753979 TI - Evaluation of quick medical reference (QMR) as a teaching tool. AB - To determine whether the Quick Medical Reference (QMR) program can improve diagnosis or enhance learning among internal medicine residents, we compared the diagnostic accuracy of the program with that of residents at various training levels. The cases were from a prospective convenience sample of 40 patients admitted by 10 first-year residents (interns) and two chief medical residents. Four sets of differential diagnoses were created for each case--the first set by an intern, the second set by a chief resident, and the third and fourth sets by QMR, using the findings of the interns and chief residents, respectively. The diagnostic accuracy of the interns and chief residents was significantly greater than that of QMR. However, the chief residents indicated that QMR did increase their understanding of disease processes and offered educational value. PMID- 9753980 TI - A freely available statistical program for testing associations. PMID- 9753981 TI - Brain wounds and their treatment in VII Corps during Operation Desert Storm, February 20 to April 15, 1991. AB - OBJECTIVE: To evaluate field neurosurgery supporting VII Corps during combat in Operation Desert Storm. RESULTS: (1) Only 1 of 22 patients who had a head wound died. (2) The one computed tomography unit in a forward hospital worked well, aiding diagnosis and surgical management. The occurrence of hematoma at a distance from the missile track has been worrisome to past field neurosurgeons, but none of 9 patients who had predebridement scans had a distant clot. (3) The number of brain wounds was fewer than expected for Americans, and the wounds were basilar in location. Iraqis, by contrast, had wounds that were randomly distributed about the head. CONCLUSIONS: (1) Although computed tomography is a useful diagnostic adjunct, its availability should not be a sine qua non for forward neurosurgery. (2) The current Kevlar helmet design appears successful. PMID- 9753982 TI - Dimensions of psychological stress in peacekeeping operations. AB - U.S. military forces are increasingly involved in a variety of multinational peacekeeping and humanitarian assistance missions. How well combat-trained units and soldiers adapt to these new roles will determine U.S. success in such operations, as well as the future health and readiness of the force. In preparing soldiers for such missions, it is critical that leaders and health care providers have a clear understanding of the nature of the stressors they are likely to encounter. This report summarizes findings from a longitudinal, descriptive case study of a U.S. Army medical unit performing a peacekeeping mission in the former Yugoslavia. The goal of the investigation was to identify key sources of stress and to delineate the effect of these stressors on the health, morale, and mental readiness of soldiers. Findings suggest a range of psychological stressors that varies somewhat across operational phases of a peacekeeping mission. Furthermore, the degree of stress experienced in various areas correlates significantly with depression, psychiatric symptoms, and low reported morale. The range of stressors is reduced and summarized in a conceptually derived model of five underlying dimensions of psychological stress salient to soldier adaptation in peacekeeping operations: isolation, ambiguity, powerlessness, boredom, and danger/threat. This model provides a useful heuristic for organizing thinking about stress in peacekeeping operations and leads to several recommendations for "countermeasures" that organizational leaders can take to maintain soldier psychological readiness during peacekeeping operations. PMID- 9753983 TI - Aggressive and impulsive behavior in military psychiatric inpatients. AB - OBJECTIVE: To determine if a history of aggressive/impulsive behavior adversely affected the return to full duty of a military psychiatric inpatient population. METHOD: Charts were reviewed for aggressive/impulsive behavior as indicated by self-report on a standardized admission form and by history in 211 consecutive admissions pooled from two separate 2-month intervals during a 9-month period. RESULTS: Seventy-three percent of the population was between the ages of 17 and 24 years. Sixty-eight percent of the population reported a history of at least one school suspension/expulsion, arrest, or military nonjudicial punishment (males, 74%; females, 45%). Seven percent of patients reporting aggressive/impulsive behavior were returned to full duty unconditionally, compared with 28% of patients not reporting said behavior. Eighteen percent of patients met the criteria for adult antisocial behavior. Only one patient with adult antisocial behavior was returned to full duty. CONCLUSIONS: This study found that a history of aggressive/impulsive behavior was positively correlated with substance abuse and negatively correlated with return to full military duty. PMID- 9753984 TI - Coping strategies and mental health problems in a military unit. AB - The aim of the present study was to investigate the effect of different coping styles on the development of self-reported mental health problems in a radically changing context. This was investigated, using a longitudinal design, by following soldiers from before entering service to 8 months of service. Based on their scores on the 30-item General Coping Questionnaire, soldiers were allocated to one of three groups: those whose coping styles were emotional, avoidance, or task focused. These three groups were assessed four times. The General Health Questionnaire (30-item version), Ursin's Health Inventory, and the Alcohol Use Disorders Identification Test (AUDIT) were used as dependent measures. Avoidance focused soldiers reported an increase in General Health Questionnaire scores over time. Furthermore, the avoidance-focused copers revealed higher scores on the AUDIT questionnaire as well as an increase in AUDIT scores over time. The present study showed that there was an interaction of personality variables and contextual factors involved. More specifically, young subjects with a preference for an avoidance-focused coping strategy are at greater risk of experiencing symptoms of mental health problems compared with task-focused and emotion-focused subjects when exposed to a radically changing environment. PMID- 9753986 TI - Military training at civilian trauma centers: the first year's experience with the Regional Trauma Network. AB - The Regional Trauma Network was launched in 1996 to provide trauma training opportunities for Army surgeons in the Southeast Regional Medical Command. Training directors at eight civilian level I trauma centers agreed to allow military surgeons to function at the fellowship level of responsibility for up to 30 days at a time. In the first year, 7 surgeons participated in rotations at five different centers and 13 surgeons attended nationally recognized trauma symposia. The response from participating civilian and military participants has been overwhelmingly positive as confidence and enthusiasm for treating seriously injured patients are refreshed. Significant lessons were learned in providing good clinical training experiences, administering a regional program, and measuring the costs and benefits of additional readiness training. Although the data collection processes were devised to capture both the actual and the opportunity costs of training at civilian centers, more participants are needed before a conclusive analysis can be made. A joint services effort on a regional basis and support throughout the chain of command are key to strengthening the surgical readiness training program. PMID- 9753985 TI - Operative videothoracoscopy in the surgical treatment of penetrating firearms wounds of the chest. AB - We prospectively analyzed our experience with operative videothoracoscopy (OVT) performed in a field military hospital in cases of penetrating firearms wounds of the thorax (PFAWT) sustained in Chechnya. From February to April 1996, we treated 206 wounded patients, of whom 37 (18.0%) had sustained chest injuries. PFAWT were present in 23 soldiers, accounting for 62.2% of all chest injuries. Twelve injuries were confined to the thorax, eight patients had associated injuries, and three soldiers had thoracoabdominal injuries. Nineteen patients had pleural drainage performed during medical evacuation. The thoracic injuries were right sided (17), involved bullets or shell splinters (23); were through and through (16), represented solitary wounds (19), and were associated with internal organ injuries (21). Fifteen patients had indications for OVT when they were delivered from the battle-field 1.5 to 22 hours after injury. All patients manifested signs of hemorrhagic shock and hemodynamic instability. Indications for OVT were ongoing intrapleural bleeding (6), clotted hemothorax (6), or marked air leakage (3) preventing lung inflation with the OP-02 apparatus (field modification). OVT revealed 12 lung wounds, nine of which were multiple wounds, pleural bleeding in 6 patients, clotted hemothorax in 11 patients, and foreign bodies in 5 patients. Two patients underwent thoracotomy, one for suspicion of heart injury and the second because we could not adequately visualize and control bleeding revealed at OVT to be from the intercostal artery in the left costovertebral angle. Eight of 23 patients had no indication for operative videothoracoscopy and were managed with continued pleural aspiration and drug therapy. Wedge resection of the lung using an Endo-GIA-30 stapler was necessary in two patients because of parenchymal destruction and bleeding. Evacuation of clotted blood by fragmentation and aspiration was satisfactory in all cases. Satisfactory manual suturing of selected lung injuries was obtained largely with intracorporeal knot tying. The duration of the procedures ranged from 40 to 90 minutes. No morbidity nor mortality was encountered in patients undergoing OVT. Postoperative pain was minimized by using OVT placement of catheters in the chest wall soft tissue with local administration of 2% Trimecain. Patients were able to stand in 10 to 12 hours and to walk by the end of the first postoperative day. All patients who underwent OVT were evacuated without drains by the third or fourth postoperative day, all tolerating sitting and standing positions. We conclude that early OVT in the military field hospital for continued bleeding, clotted hemothorax, and continued major air leakage has several advantages in military patients with PFAWT: early definition and management of organ injury; identification and control of bleeding in most instances; earlier and more accurate assessment for thoracotomy; vigorous lavage and removal of projectiles such as bone fragments and evacuation of clotted hemothorax; early debridement with suture closure of the thoracic wall canal; and minimal postoperative pain with dramatically reduced suppurative sequelae and bronchopleural fistulae. PMID- 9753987 TI - Statistical modeling using preoperative prognostic variables in predicting extracapsular extension and progression after radical prostatectomy for prostate cancer. AB - OBJECTIVE: To predict the risk of extracapsular extension and postoperative recurrence before radical prostatectomy (RP) for prostate cancer. METHODS: We performed multivariate Cox regression analysis on preoperative variables in 260 clinically localized prostate cancer patients who underwent RP. With these data, we constructed a relative risk of recurrence (Rr) equation and an equation to predict the probability of extracapsular extension (PECE) before RP. RESULTS: Rr is calculated as exp[(0.47 x race + 0.14 x PSAST) + (0.13 x worst biopsy Gleason sum) + (1.03 x stage T1c) + (1.55 x stage T2b,c)], where PSAST indicates a sigmoidal transformation of prostate-specific antigen. PECE is calculated as 1/[1 + exp(-Z)], where Z = -2.47 + 0.15 (PSAST) + 0.31 (worst biopsy Gleason sum) + 0.18 (race) + 0.16 (stage T1c) + 0.38 (stage T2b,c). CONCLUSION: These two equations can be used preoperatively to predict the probability of extracapsular disease and the risk of prostate-specific antigen recurrence in patients undergoing RP. PMID- 9753988 TI - Update on wound healing: a review of the literature. PMID- 9753989 TI - The hidden cost of tobacco smoke. PMID- 9753990 TI - Perceptions of current and recent military internal medicine residents on operational medicine, managed care, graduate medical education, and continued medical service. PMID- 9753991 TI - Effects of volume control, pressure control, and pressure-regulated volume control on cardiopulmonary parameters in a normal rat lung. AB - The purpose of this study was to investigate the differences in diaphragm shortening and cardiopulmonary parameters at varying tidal volumes during volume control (VC), pressure control (PC), and pressure-regulated volume control (PRVC). A miniaturized ultrasonic sensor attached to the inferior surface of the upper costal surface of the right hemidiaphragm of 16 Sprague-Dawley rats provided a direct assessment of diaphragm shortening. Within each control mode of mechanical ventilation, the tidal volume was increased from 3 to 12 ml in increments of 3 ml. There were no differences in cardiac output, mean arterial pressure, central venous pressure, peak inspiratory pressure, or end-tidal CO2 among the three modes of mechanical ventilation. At equivalent tidal volumes, diaphragm shortening was less during PRVC than during VC or PC. This finding suggests that differences in diaphragm shortening may be caused by shorter resting (end-expiratory) diaphragm muscle length. The cardiopulmonary data obtained in this study provide new information for clinicians to consider when using various modes of ventilation, particularly PRVC. PMID- 9753992 TI - A cost-benefit analysis of a routine varicella vaccination program for United States Air Force Academy cadets. AB - The United States Air Force Academy (USAFA) is one of the nation's military universities, with the mission to educate and motivate cadets to be career Air Force officers. This diverse population arrives at the USAFA with varying immunization records and disease histories. A review of USAFA cadet medical records identified an alarming cost of treating a simple, preventable, generally childhood disease: chickenpox. In July 1995, a cost-benefit analysis was performed on the use of varicella vaccine among cadets and preparatory school students at the USAFA. Based on this analysis, the USAFA implemented a strategy of serologic screening and vaccination. Although this study does not establish causation, follow-up data showed a dramatic decrease in cases, associated hospitalizations, and therefore costs during the first year after implementation. Fiscal projections indicate that these costs savings should increase through year 4 of the program and continue thereafter. At year 4, the total cadet population will have been serologically screened and/or vaccinated against chickenpox. PMID- 9753993 TI - From atabrine in World War II to mefloquine in Somalia: the role of education in preventive medicine. AB - Preventive medicine requires an understanding not only of the disease to be prevented, but a complete understanding of conditions affecting the disease transmission, human nature, and the historic situation of the target group. An analysis of a new drug introduction (Atabrine) during World War II is viewed from multiple perspectives and is compared with the introduction of mefloquine during the mission to Somalia 50 years later. Common themes of educational failure at the end-user and policy-marker levels are shown as barriers to effective preventive medicine efforts. PMID- 9753994 TI - Information technologies for Marine Corps combat medicine. AB - Future Marine Corps warfighting concepts will make it more difficult to locate casualties, which will complicate casualty evacuation, lengthen casualty wait times, and require infantrymen or corpsmen to provide more extensive treatment. In these future scenarios, information flow and communications will be critical to medical functions. We asked, for Navy medical support to the Marines, what information will future combat medicine require and what technologies should supply those information needs? Based on analyses of patient data streams, focus groups of Navy medical personnel, and our estimates of the cost and feasibility of communications systems, we recommend the following: (1) increase medical training for some fraction of Marines, especially in hemorrhage control; (2) augment corpsmen's training; (3) furnish data systems for evacuation and supply that would provide in-transit visibility and simplify requests; (4) provide all ground medical personnel with access to treatment information systems and limited voice communications; and (5) exploit e-mail systems to reduce reliance on voice communications. Implementation time frames are discussed. PMID- 9753995 TI - Speech discrimination in the sensorineural hearing loss patient: how is it affected by background noise? AB - There are patients who have normal hearing below 3,000 Hz and normal speech discrimination who still complain of hearing difficulty, especially when background noise is present. The objective of this study is to document the fact that these individuals have a significant hearing impairment that is not detected with routine testing. We retrospectively reviewed 67 audiograms selected for 50 dB loss or greater at 3, 4, 6, and 8 kHz and speech discrimination scores better than 80%. Patients in this group had also previously undergone speech discrimination testing in the presence of 50-dB calibrated cafeteria noise at the time of the initial audiogram. Identical testing was carried out on 48 control subjects without hearing loss. The speech discrimination scores of the hearing impaired group were lower than in the control group when tested in a quiet booth (88.2 and 98.2%, respectively). The significant finding was the change in the speech discrimination score when tested in noise. We found that the study group had a 33.1% loss in speech discrimination when tested in the presence of background noise. The control group had only a 5.2% loss in speech discrimination in the presence of the same noisy background; this was statistically different (p = 0.001). Our conclusion is that patients complaining of hearing loss who have normal low- to mid-frequency hearing and good speech discrimination should be tested in the presence of noise to adequately document their degree of impairment. Our findings also support the theories of signal attenuation and secondary auditory pathway distortions as causes of the loss of speech discrimination. PMID- 9753996 TI - Bilateral total knee replacement for traumatic arthritis from blast injury in Vietnam. PMID- 9753997 TI - Axillary artery aneurysms. AB - Aneurysms of the axillary artery are rare but potentially dangerous lesions that threaten the upper extremity with vascular and neurologic compromise. Most can be treated effectively with surgical excision and vascular grafting. An illustrative case is presented. These aneurysms may arise as pseudoaneurysms secondary to trauma or iatrogenic complications, or as degenerative lesions often secondary to the chronic use of crutches. They may also arise as postobstructive lesions in patients with thoracic outlet syndrome. Signs and symptoms vary with the cause of the aneurysm and may include mass effects with brachial plexus compression and thromboembolic events involving the hands and fingers. Arteriography is the mainstay of diagnosis, and treatment should be considered in most of these lesions as soon as they become apparent to prevent limb loss or dysfunction. PMID- 9753998 TI - Echinococcal disease and the provision of humanitarian surgical aid in Bosnia. AB - The Canadian Armed Forced have been deployed to the republics of the former Yugoslavia since 1992 as part of the United Nations Protection Force and the NATO led Implementation Force. Most of the combat arms units have been supported by small, self-contained surgical teams for essential surgery. Considerable benefit is gained by cooperating with civilian surgeons. The experience of treating five patients with complicated hydatid disease endemic to the area is examined. Treatment of these patients requires performing major surgical procedures under austere conditions and must be undertaken with care. Careful selection and communication with the surgical nursing team and civilian surgeons is essential. Well selected cases can also pay tremendous dividends in terms of maintaining the skills of personnel who must be prepared for any emergency in addition to providing vital surgical assistance to these patients. PMID- 9753999 TI - [Gamma-knife radiosurgery for metastatic brain tumors from primary lung cancer]. AB - Forty patients with metastatic brain tumors from primary lung cancer underwent radiosurgery (gamma-knife). We retrospectively compared their prior treatment history number of metastatic foci, and performance status, to evaluate the effects of, and indications for, gamma-knife therapy. After both the primary and the metastatic tumors were controlled, performance status could be used as an index in the choice of gamma-knife therapy. Our results demonstrate that repeated gamma-knife radiosurgeries prolonged survival time. Gamma-knife radiosurgery improves quality of life and prognosis of patients with metastatic brain tumors. PMID- 9754000 TI - [Value of transtracheobronchial ultrasonography in the assessment of mediastinal lymphadenopathy in patients with lung cancer]. AB - In patients with lung cancer, decisions regarding treatment can depend on the diagnosis of hilar and mediastinal nodal involvement. We prospectively compared the diagnostic value of computed tomography (CT) with that of transtracheobronchial ultrasonography (TUS) in the evaluation of lymphadenopathy. Five patients with resectable lung cancer were studied. TUS was done with EU-M 20 or M 30 and lymph nodes located at #3, #4, #7, ipsilateral #10, and #11 were observed and measured. TUS findings, CT findings, and histological findings were evaluated and compared. The sizes of lymph nodes as measured by TUS were similar to or slightly smaller than their sizes as measured by CT. Hilar lymph nodes and lymph nodes located at right #4 were clearly observed with TUS, but were sometimes unclear with CT. Diagnosis of model involvement by TUS needs further study. PMID- 9754001 TI - [Behcet's disease with acute inflammatory foci and localized secondary organizing pneumonia]. AB - A 68-year-old man with incomplete-type Behcet's disease was admitted to our hospital with a one-month history of a high fever and a persistent pulmonary infiltration shadow in the left lower lobe. Chest CT films showed localized round air-space consolidation in the left lower lobe. The fever did not responded to antibiotics, but did resolve with administration of prednisolone 15 mg/day. Left lower lobectomy was done to avoid life-threatening hemoptysis. Histological examination of the main pulmonary lesions revealed old vascular changes such as irregular intimal thickening and perivascular fibrosis. Disoriented elastic fibers of arteries was seen in some areas as were acute inflammatory foci with neutrophilic infiltration, and secondary organizing pneumonia with Masson bodies. From these findings Becet's disease related pulmonary involvement could not be excluded. Localized secondary organizing pneumonia associated with acute inflammatory foci, as found in this case, is a very rare pulmonary manifestation of Behcet's disease. PMID- 9754002 TI - [Chronic necrotizing pulmonary aspergillosis resistant to antimycotic drugs and treated by partial pulmonary resection]. AB - A 46-year-old man with a history of left upper lobectomy for pulmonary tuberculosis was admitted to our hospital because of dilated cardiomyopathy. During hospitalization, fever and weight loss developed. The cause was suspected to be a round mass inside a cavity and a neighboring infiltrative shadow in the left upper lung field as seen on chest radiography. A percutaneous needle biopsy was done, and examination of the specimen showed an aggregate of Aspergillus fumigatus hyphae. Fluconazole (FCZ) was injected through an intracavitary catheter every day, and was then given by mouth. Treatment with FCZ was effective temporarily. However, he was again admitted to our hospital because of lower extension of the cavity and deteriorated inflammatory findings. From the clinical course, chronic necrotizing pulmonary aspergillosis was diagnosed. Treatment with all available antifungal agents did not improve his condition. Although he had decreased cardiac function due to dilated cardiomyopathy, partial pulmonary resection was done. The cavity with the fungus ball was resected completely. As of the time of this writing, he remains free of aspergillosis. PMID- 9754004 TI - [Ramsay Hunt syndrome associated with eosinophilic pneumonia]. AB - A 66-year-old woman was admitted to the hospital because of dry coughing. Ten days before admission, the patient had suffered from facial palsy accompanying otic zoster infection (Ramsay Hunt syndrome). Acyclovir was given, and during the two weeks after admission, the facial palsy resolved completely. The dry coughing worsened, and marked eosinophilia developed (1.930/mm3). A chest roentgenogram and a computed tomogram revealed wandering non-segmental infiltration in the left lung field. Examination of a specimen obtained by transbronchial lung biopsy revealed moderate eosinophilic infiltration into thickened alveolar septa and alveolar spaces. An elevated CD 4/CD 8 ratio (4.12) and a high level of eosinophilic cationic protein (8.730 micrograms/l) were found in bronchoalveolar lavage fluid. Eosinophilic pneumonia was diagnosed. The patients condition improved without medication within one month after the onset of the dry coughing. Laboratory results revealed no parasitic or mycotic infection, and both an acyclovir skin test and a lymphocyte stimulation test were negative, which suggested that the pneumonia had been induced by an allergic reaction to unknown antigens resulting from Th 1/Th 2 imbalance after reactivation of varicella zoster virus latent in sensory ganglia. PMID- 9754003 TI - [Pulmonary thromboembolism that developed during an airplane flight "economy class syndrome"]. AB - The occurrence of thromboembolic phenomena during long-duration airplane flights is called "economy-class syndrome". Recently it has become more popular for Japanese to go abroad by airplane, and an increase in the prevalence of pulmonary thromboembolism should be expected. However, there are few reports of the economy class syndrome in Japan. A 52-year-old woman was admitted to our hospital because of chest discomfort and dyspnea that developed during an airplane flight. We suspected pulmonary thromboembolism, on the basis of a chest X-ray film and on electrocardiogram. A ventilation-perfusion lung scan disclosed mismatching between ventilation and perfusion in the right upper lung field. Pulmonary thromboembolism was confirmed by pulmonary arteriography. The patient was treated with heparin and urokinase. A phlebogram of the legs showed no significant findings. There was no history of thromboembolic disease or of consumption of oral contraceptives. We conclude that the pulmonary thromboembolism might have been caused by stasis of blood in the lower limb veins during the airplane flight. We emphasize the importance of including pulmonary thromboembolism in the differential diagnosis of patients with chest discomfort and dyspnea that develop during airplane flights. No noninvasive test can lead to a definitive diagnosis of pulmonary thromboembolism. Early pulmonary angiography should be recommended when pulmonary thromboembolism is suspected. PMID- 9754005 TI - [Procainamide-induced lupus in a patient with bilateral pleural effusion]. AB - A 70-year-old physician was admitted to our hospital because of bilateral pleural effusion and left-sided chest pain on deep inspiration. On admission, the APTT was prolonged and was not corrected with a 1:1 mixture of normal plasma. Results of serological examinations included a positive lupus-anticoagulant test and a positive ANA test at a titer of 1:1,280 in a homogeneous pattern. The patient's age, sex, symptoms, signs, and laboratory results all argued against the diagnosis of SLE except for ANA and lupus anticoagulant test. Because procainamide had been prescribed (250 mg every 6 h) for premature ventricular contractions for eight years before admission, procainamide-induced lupus was suspected. Procainamide was discontinued. Chest pain persisted and tests for c reactive protein were positive. Prednisolone was administered. Procainamide induced lupus was diagnosed, because anti-histone H 2 A-H 2 B complex antibodies were high by enzyme-linked immunosorbent assay, and IgM-class anti-histone antibodies were found in response to H1, H 2 B and H 2 A-H 2 B complex (immunoblotting), which suggested the drug induced lupus. There are only a few reports of drug induced lupus in which the lupus-anticoagulant test was positive and prednisolone was indicated. The measurements of anti-histone antibodies and of expression of anti-histone antibodies were useful in distinguishing drug induced lupus from SLE. PMID- 9754006 TI - [Small cell lung cancer associated with nephrotic syndrome: remission after chemotherapy]. AB - A 66 year-old man was found to have pointed out a 1-cm tumor shadow, on a chest X ray film when he underwent a gastrectomy because of advanced gastric cancer. Five months after the operation, edema and proteinuria developed, and a chest X-ray film revealed enlargement of the tumor. There was no sign of recurrence of the gastric cancer. Nephrotic syndrome due to IgA-nephropathy and small cell lung cancer was diagnosed. Chemotherapy (carboplatin and etoposide) was effective against both the lung tumor and the nephrotic syndrome. Small cell lung cancer may have been involved in the pathogenesis of the nephrotic syndrome in this patient. PMID- 9754007 TI - [Interstitial pneumonitis detected by bronchoalveolar lavage and transbronchial lung biopsy in adult-onset Still's disease]. AB - A 44-year-old woman was admitted to our hospital because of a high fever that had continued for three weeks. She complained of a sore throat and arthralgia, and had a salmon-pink rash, lymphadenopathy, liver dys-function, and hyperferritinemia. Tests for RF and ANA were negative. Adult-onset Still's disease was diagnosed. Despite administration of steroids, pericarditis, interstitial pneumonitis, and disseminated interavascular coagulation developed. After cyclophosphamide was given, the patient's condition improved, but reticular shadows and volume loss remained on the chest X-ray film. A chest CT scan showed ground-glass-like opacities and linear shadows, and irregular bronchovascular bundles. Bronchoalveolar lavage and transbronchial lung biopsy were done. Alveolar macrophages accounted for 71% of the cells in the bronchoalveolar lavage fluid, and lymphocytes (CD 4/ CD 8 ratio = 1.01) accounted for 29%. Examination of a specimen obtained by transbronchial lung biopsy showed thickened alveolar walls and infiltration of lymphocytes. Reports of cases of adult-onset Stills disease that include results of bronchoalveolar lavage and transbronchial lung biopsy are rare. PMID- 9754008 TI - [Pulmonary disease after massive inhalation of Aspergillus niger]. AB - A 60-year-old man was admitted to the hospital because of fever, coughing, and dyspnea that developed after he entered a silo that had been filled with chips of wood in the preceeding 3 months. A chest X-ray film revealed bilateral ground glass shadows. Histologic study of the lung showed a multifocal acute process; the alveoli and interstitial areas contained many fungal hyphae and spores. Cultures from both bronchoalveolar-lavage-fluid and the chips in the silo revealed Aspergillus niger. Serologic reactions were negative to 10 antigens known to induce hypersensitivity pneumonitis. Furthermore, the patient's serologic reaction to the extracts of fungi obtained from the bronchoalveolar lavage-fluid was negative. The patients recovered quickly without steroid therapy. We believe that this patient's diseases was "organic dust toxic syndrome". PMID- 9754009 TI - [Sarcoidosis presenting as a diffuse micronodular shadow on a chest radiograph]. AB - A 36-year-old man was admitted to our hospital with chest-radiographic findings of diffuse ground-glass shadows in both lungs. A chest CT scan revealed disseminated micronodules with mediastinal lymphadenopathy. Histological examination of a transbronchial lung-biopsy specimen showed non-caseous epithelioid cell granulomata. Marked lymphocytosis and an abnormally high CD 4/CD 8 ratio was found in bronchoalveolar lavage fluid. These findings suggested a diagnosis of sarcoidosis despite the atypical radiographic findings. PMID- 9754010 TI - [Pulmonary dirofilariasis associated with pleural effusion]. AB - A 61-year-old man was admitted to our hospital because of a long history of productive coughing. A chest roentgenogram and CT scan showed a right-sided pleural effusion. The effusion fluid was blood-stained but showed no cytological evidence of malignancy. Marked eosinophilia was found in blood and in the pleural effusion fluid. Ouchterlony's double-diffusion test done with the patients serum and pleural effusion fluid in agarose showed specific bands toward Dirofilaria immitis antigen, and this specificity was confirmed with an enzyme linked immunosorbent assay inhibition test. The final diagnosis was pulmonary dirofilariasis, and the patient responded to diethylcarbamazine. PMID- 9754011 TI - [Pulmonary sclerosing hemangioma with specific CT findings]. AB - A 44-year-old woman was admitted to our hospital for evaluation of an abnormal lung shadow. Chest computed tomography (CT) revealed a tumor surrounded by air space and an infiltrative shadow in the right S2. Right upper lobectomy was performed and pulmonary sclerosing hemangioma was diagnosed. Usually, pulmonary sclerosing hemangioma shows a solitary round nodule on a chest CT scan. We report a case of pulmonary sclerosing hemangioma with an unusual shadow on a chest CT scan, and review the literature. PMID- 9754013 TI - [Diminution of the number of gamma delta T lymphocytes in hepatocellular carcinoma patients treated with transcatheter arterial embolization]. AB - gamma delta T lymphocytes, which are CD3+ lymphocytes that express gamma delta chains of the T-cell antigen receptor (TCR) on their surface, are functionally distinct from alpha beta T lymphocytes, which express alpha beta chains of the TCR. gamma delta T lymphocytes are thought to differentiate in mouse hepatic sinusoids, to play a role in antitumor action, and to act as natural killer cells. The purpose of this study was to examine whether gamma delta T lymphocytes in the peripheral blood are suppressed when hepatic sinusoids are damaged during transcatheter arterial embolization (TAE). The numbers of alpha beta T lymphocytes and gamma delta T lymphocytes in the peripheral blood were examined with monoclonal antibodies and flow cytometry before and after TAE in 32 patients (from 46 to 78 years of age) with liver cirrhosis and hepatocellular carcinoma. The number of alpha beta T lymphocytes before and after TAE was unchanged. However, the number of gamma delta T lymphocytes and the ratio of gamma delta T lymphocytes to CD3+ lymphocytes were significantly decreased for 3 weeks after TAE treatment. This decrease suggests that TAE suppresses the supply of gamma delta T lymphocytes to the peripheral blood. In addition, TAE may weaken a patient's antitumor immunity, because gamma delta T lymphocytes that have antitumor activity decrease after TAE. PMID- 9754014 TI - [The mechanism of donor specific unresponsiveness in renal transplant recipients]. AB - A study was conducted to clarify the mechanism of donor specific immunological unresponsiveness in renal transplant recipients with a well functioning kidney, since this might provide a clue to the nature of donor specific immunosuppression. Peripheral blood lymphocytes (PBL) were obtained from three renal transplant recipients, the donors (mother), the fathers and healthy volunteers as 3rd party and used in the present study. PBL from one of the three renal transplant recipients, which were donor specific unresponsive in MLR (Mixed Lymphocyte Reaction) and CML (Cell Mediated Lympholysis), suppressed the MLR, CML and synthesis of IL-2 from 3rd party PBL in a double chamber assay only when stimulated by the donor PBL. This suppression was abolished by the treatment with CD3 or CD8 antibodies and complement. RT-PCR was performed to investigate the suppressive effect of the recipient's PBL on IL-2 mRNA expression. In the double chamber MLR, expression of IL-2 mRNA by the PBL from 3rd party was also suppressed only when the recipient's PBL were stimulated by the donor PBL. These results indicate that one of the mechanisms responsible for donor specific immunological unresponsiveness is the presence of donor specific suppressor T cells in recipient's PBL and that these cells project suppressive effect on lymphocytes by producing a humoral suppressive factor(s) that suppress the expression of the IL-2 mRNA only when stimulated by donor PBL. PMID- 9754015 TI - [An autopsy case of rheumatoid arthritis accompanied with acute exacerbation of interstitial pneumonia]. AB - An autopsy case of rheumatoid arthritis (RA) with acute exacerbation of interstitial pneumonia is reported. A 57-year-old woman with longstanding RA was admitted to our hospital because of progressive dyspnea. On chest roentogenogram, diffuse interstitial shadow was confirmed in both lungs. Chest computed tomography (CT) showed diffuse lesion of elevated density of CT level in both lung. She was diagnosed as an acute exacerbation of interstitial pneumonia, and treated by methylpredonisolone pulse therapy (1,000 mg/day). Although cyclosporin A (2 mg/kg/day) was combined to steroid therapy, she was died of progressive respiratory failure. The histological findings of the lung showed extensive fibrosis with alveolar damage associated with hyaline membranes, edema and hemorrhage in alveolar space. PMID- 9754016 TI - [Interferon-alpha treatment for chemotherapy-resistant primary macroglobulinemia with stomach and lung invasion]. AB - We reported a case of primary macroglobulinemia with stomach and pulmonary invasion. The patient was 71 years-old who had cervical lymphadenopathy and abdominal pain. Biopsy material of cervical lymph node showed non-Hodgkin's lymphoma, and he was diagnosed primary macroglobulinemia by IgM immunological histo-chemical staining of materials of stomach biopsies. Combination chemotherapies were not effective for the reduction of IgM-lambda protein, and organ invasion seemed to be progressive, so we tried interferon-alpha (IFN-alpha) to control M component. Daily injection of 6 megaunits of IFN-alpha induced significant reduction of M component and pulmonary invasion. This favorable changes were observed for 1 year. However, his pulmonary invasion on X-ray films relapsed and he died of respiratory failure by reason of severe pneumonia. IFN alpha is currently available for myeloproliferative disease, especially chronic myelogenous leukemia and multiple myeloma. This case report showed that IFN-alpha was also available for primary macroglobulinemia. PMID- 9754017 TI - [Proliferation of corneal epithelial cells in diabetic rats]. AB - In order to elucidate the mechanisms of diabetic corneal epitheliopathy, we investigated the proliferation of corneal epithelial cells of streptozotocin induced diabetic rats in the 1st, 2nd, 3rd and 6th months after the onset of diabetes using 3H-thymidine autoradiography. After the 1st month in diabetic rats, histological examination revealed that corneal epithelial layer was thin, and proliferation of corneal epithelial cells was decreased compared with that of normal untreated rats. After the 2nd month, proliferation of corneal epithelial cells showed no difference from normal rats. After the 3rd month in diabetic rats, the attachment of the epithelial layer to the stroma was loose and proliferation of corneal epithelial cells was increased compared with that of normal untreated rats. After the 6th month, proliferation of corneal epithelial cells showed no difference between diabetic rats and normal rats. We suggest that increased turnover rate of corneal epithelial cells may account for increased proliferation of corneal epithelial cells in diabetic rats during the 3rd month. PMID- 9754018 TI - [The correlation between gap junction-mediated communication and cell proliferation among retinal pigment epithelial cells]. AB - I investigated gap junction-mediated intercellular communication during cell proliferation of retinal pigment epithelium (RPE) following experimental retinal tear formation in albino rabbit eyes. I used a dye-coupling method with lucifer yellow CH. I also evaluated the expression of proliferating cell nuclear antigen (PCNA) in the RPE cells. Immediately after tear formation, lucifer yellow CH introduced into a target RPE cell appeared in neighboring cells. The number of dye-recipient cells was 33.8 +/- 4.7 (mean +/- standard error of the mean). One week later, the number of fluorescent RPE cells decreased to 2.3 +/- 0.9. The extent of dye transfer between RPE cells increased (17.5 +/- 4.1). There was no expression of PCNA in RPE cells immediately after tear formation, but it became apparent as the nuclei of RPE cells became positive for PCNA. The immunoreactive cells and cell population decreased one mouth later. Because the duration of decreased dye-coupling coincides with that of cell proliferation, these finding suggest that an alteration of gap junctional intercellular communication is involved in RPE cell proliferation in the present experimental setup. PMID- 9754019 TI - [Disorders of choroidal circulation in diabetic maculopathy]. AB - Diabetic maculopathy is classified into 3 types, namely, macular edema, ischemic maculopathy, and pigment epitheliopathy. Blood-retinal barrier disturbance and the influence of the posterior vitreous membrane have been reported as the cause of diabetic maculopathy. However, its association with the choroidal circulation feeding the outer layer of the retina which involves the outer blood-retinal barrier has not been clarified yet. In this work, we studied the presence of the choroidal circulatory disturbance by performing indocyanine green angiography (IA) on patients with diabetic maculopathy. Choroidal circulatory disturbance was also differentiated from fluorescent block with a scanning laser ophthalmoscope (SLO). The proportion of hypofluorescence observed in IA was 4 eyes in 2 patients among 37 eyes in 24 patients (11%) in the macular edema group, 3 eyes in 2 patients among 14 eyes in 10 patients (21%) in the ischemic maculopathy group, and 26 eyes in 17 patients among 33 eyes in 22 patients (79%) in the pigment epitheliopathy group. Of the cases showing hypofluorescence in IA in the pigment epitheliopathy group, 4 eyes in 4 patients had hypofluorescence due to both choroidal circulatory disturbance and fluorescent block. This suggests that the choroidal circulation disturbance is partly involved in diabetic maculopathy mainly in the pigment epitheliopathy group. PMID- 9754020 TI - [Dielectric dispersions of alpha, beta H, beta L and gamma crystallin solutions]. AB - The crystallins which constitute the major soluble proteins of the lens are known to change into insoluble proteins in aged or cataractous lenses. Such changes would affect protein-bound water and bulk water around the crystallin molecules. We measured the a.c. admittances of several kinds of crystallin, solutions in the frequency range from 10 kHz to 2 GHz to define the passive electrical properties of bound water. With the experimental data, we calculated the relative permittivity, conductivity, and loss factor. The admittance of crystallin solutions had essentially three dielectric dispersions and we could divide them into three dependent dispersions by a curve-fitting analysis based on a three term Cole-Cole equation. The lowest frequency dispersion around a few MHz was due to protein particles, the middle frequency of the 10-100 MHz range to bound water, and the highest frequency of over 1 GHz to bulk water. The dielectric behavior of crystallin molecules and bound water depended on the protein concentration. With the increase in molecular weight of crystallins, the dielectric increments of both dispersions increased and the characteristic frequencies of the dispersions decreased significantly. PMID- 9754021 TI - [Association between watershed zone and visual field defect in normal tension glaucoma]. AB - In order to evaluate the association between the watershed zone and glaucomatous optic damage, we performed indocyanine green fluorescence angiography with a scanning laser ophthalmoscope in 54 eyes of 27 patients with normal tension glaucoma. The visual field indices were measured with a Humphrey Field Analyzer. We identified 8 eyes (14.8%) of 7 patients with a watershed zone not including the optic nerve head (type I), 32 eyes (59.3%) of 20 patients with the zone partially including the optic nerve head (type II), and 14 eyes (26.0%) of 10 patients with the zone including the optic nerve head (type III). Of the total of 27 patients, 10 patients (37.0%) had different types in each eye. In these patients, the mean deviation (MD) of visual field indices was worse in the eye with the watershed zone which included a larger part of the optic disc than in the contralateral eye (p < 0.05). Conversely, the eye with worse MD than the contralateral eye had a watershed zone which included a larger part of the optic disc than the other eye (p < 0.05). The location of watershed zone appeared to influence the progression of the visual field defect. PMID- 9754022 TI - [The diagnostic significance of polymerase chain reaction for ocular samples in viral retinitis]. AB - During the past two years we studied the incidence of infection by eight members of the herpesvirus family in ocular samples (tear fluid, the aqueous, and the vitreous) using the polymerase chain reaction (PCR). A total of 31 ocular samples were collected from 22 eyes of 13 patients. The series comprised five cases of acute retinal necrosis, four of cytomegalovirus retinitis, three of proliferative diabetic retinopathy, and one of ocular malignant lymphoma. In 12 eyes of 9 patients with viral retinitis, causative viral DNA was detected either from the tear fluid (1/12, 8%), the aqueous (6/7, 86%), or the vitreous (1/1, 100%). In the fellow eyes free of viral retinitis, no viral DNA was detected in the samples (6 of tear fluid and one of the aqueous). Out of the other 4 patients without viral retinitis, viral DNA was detected in one patient with ocular malignant lymphoma. The findings show that PCR is a useful and sensitive method in the diagnosis of viral retinitis, but that it may give false positive results. The aqueous and the vitreous samples gave more positive results than tear fluid. PMID- 9754023 TI - [Treatment of cytomegalovirus retinitis in AIDS with an intraocular sustained release ganciclovir implant]. AB - Active cytomegalovirus (CMV) retinitis in patients with acquired immunodeficiency syndrome (AIDS) was treated with an intraocular sustained-release ganciclovir implant. A total number of 19 implants were performed in 15 eyes of 9 AIDS patients. The intraocular sustained-release ganciclovir was effective in preventing reactivation of CMV retinitis in 15 of the 19 implants, ineffective in 3, and undetermined in 1. All ineffective cases had been resistant to ganciclovir therapy before the implants. Vision after the therapy was maintained at better than 0.5 except for one eye. There were no serious ocular complications caused by the therapy. Among 5 patients with unilateral CMV retinitis, 2 unaffected eyes developed CMV retinitis during this therapy. In addition, another patient developed presumed CMV infection in other systemic organs. Based on these data, the intraocular sustained-release ganciclovir implant was considered to be useful for the treatment of CMV retinitis in AIDS. PMID- 9754024 TI - [Preventive effects of diclofenac ophthalmic solution on post-cataract-surgical cystoid macular edema]. AB - Using eyes undergoing phacoemulsification followed by implantation of a foldable acrylic intraocular lens (IOL) designed for small-incision cataract surgery, a multi-center study was performed to compare a non-steroidal ophthalmic solution (0.1% diclofenac) to a steroidal ophthalmic solution (0.1% fluorometholone) in preventing cystoid macular edema (CME) and inducing disruption of the blood aqueous barrier determined by laser flare cellmetry. The incidence of CME, noted in 3 out of 53 eyes (5.7%) receiving diclofenac and in 29 out of 53 eyes (54.7%) receiving fluorometholone, was significantly lower in the diclofenac group. The flare in the anterior chamber was also significantly lower in the diclofenac group; when compared between eyes with and without CME, the amount of flare was significantly higher in the former group. These findings suggest that diclofenac effectively prevents CME following cataract surgery and that CME is closely related to the breakdown of the blood-aqueous barrier. PMID- 9754025 TI - [Immunohistochemical studies on factors involved in after cataract]. AB - In this study, lenses of autopsy eyeballs, anterior capsules including lens epithelium taken during operation for cortical cataract, after cataract tissue obtained at the time of operation, and Elschnig's pearles and Soemmerring's ring from autopsy eye-balls were examined for a variety of factors, such as growth factors, cytokines, bioactive substance factors, cytoskeleton proteins and extracellular matrices by immunocytohistochemistry. Preoperative lens epithelium expressed epidermal growth factor (EGF), EGF-receptor (R), fibroblast growth factor (FGF), FGF-R, interleukin (IL)-1-RII, tumor necrosis factor-alpha (TNF alpha), plasminogen activator inhibitor type-1 (PAI-1), keratin and laminine. In addition to the above factors, opacified fibrous capsule in after cataract expressed transforming growth factor-beta (TGF-beta), insulin-like growth factor II (IGF-II), platelet derived growth factor-AB (PDGF-AB), IL-6, prostaglandin-E2 (PG-E2), alpha smooth muscle actin, fibronectin, and I and III to VI type collagen. Elschnig's pearls expressed FGF-R, TNF-alpha, and laminin. Soemmerring's ring expressed EGF, FGF, FGF-R, IL-1-RII, keratin, tissue-PA, and PAI-1. PMID- 9754026 TI - [Histological studies in the developing eyelids of congenital microphthalmic (Cts) mice]. AB - The Cts mouse is a mutant of the Jcl:ICR strain with congenital cataract and small eyes in homozygotes. In the present study, attempts were made to measure postnatally the palpebral fissure and to examine the process of eyelid development by light microscopy in both normal (+/+) and homozygous (Cts/Cts) mice. The width of the palpebral fissure in homozygotes was significantly smaller than in unaffected mice. To investigate the process of eyelid development, tissue specimens from normal and Cts homozygous mice were prepared with hematoxylin and eosin, and observed by light microscopy on days 14, 16, and 18 of gestation and on days 0, 7, and 14 after birth. In both groups, the eyelid fused on day 16 of gestation and opened completely on day 14 after birth. The primordium of the orbicularis oculi muscle was defined on day 16 of gestation, and the primordia of hair follicles and Meibomian glands were detected on day 18 of gestation. In the present study, there were no significant differences in the process of eyelid development between normal and homozygous mice. Eyelid development has thus far been thought to be self-determining and not affected by tissue interaction, and the present findings support this line of thought. PMID- 9754027 TI - [Detection of parafoveal scotoma by multifocal electroretinograms]. AB - We investigated the relation between multifocal electroretinograms (M-ERGs) and artificial parafoveal scotoma. M-ERGs were recorded from normal subjects using a circular piece of black paper attached to a monitor. Lower response density around the 10 to 15 degree parafovea region was not observed up to 3 degree scotoma (visual angle), but was detected above 5 degree scotoma in field topography of M-ERGs. The shape of the scotoma in field topography was not circular but somewhat oval. The results from two cases of parafoveal retinal degeneration were in good accordance with this basic study in normal subjects. We proved that detection of parafoveal scotoma by M-ERG is limited in comparison with the results obtained by automated static perimetry. PMID- 9754028 TI - Methionine ligation strategy in the biomimetic synthesis of parathyroid hormones. AB - In biological systems, both proteolysis and aminolysis of amide bonds produce activated intermediates through acyl transfer reactions either inter- or intramolecularly. Protein splicing is an illustrative example that proceeds through a series of catalyzed acyl transfer reactions and culminates at an O- or S-acyl intermediate. This intermediate leads to an uncatalyzed acyl migration to form an amide bond in the spliced product. A ligation method mimicking the uncatalyzed final steps in protein splicing has been developed utilizing the acyl transfer amide-bond feature for the blockwise coupling of unprotected, free peptide segments at methionine (Met). The latent thiol moiety of Met can be exploited using homocysteine at the alpha-amino terminal position of a free peptide for transthioesterification with another free peptide containing an alpha thioester to give an S-acyl intermediate. A subsequent, proximity-driven S- to N acyl migration of this acyl intermediate spontaneously rearranges to form a homocysteinyl amide bond. S-methylation with excess p-nitrobenezensulfonate yields Met at the ligation site. The methionine ligation is selective and orthogonal, and is usually completed within 4 h when performed at slightly basis pH and under strongly reductive conditions. No side reactions due to acylation were observed with any other alpha-amines of both peptide segments as seen in the synthesis of parathyroid hormone peptides. Furthermore, cyclic peptide can also be obtained through the same strategy by placing both homocysteine at the amino terminus and the thioester at the carboxyl terminus in an unprotected peptide precursor. These biomimetic ligation strategies hold promise for engineering novel peptides and proteins. PMID- 9754029 TI - Small structural ensembles for a 17-nucleotide mimic of the tRNA T psi C-loop via fitting dipolar relaxation rates with the quadratic programming algorithm. AB - Solution structures are typically average structures determined with the help of nmr-derived distance and torsion angle information. However, when a biomolecule populates significantly different conformations, the average structure might be prone to artifacts, and other refinement strategies are necessary. For example, when experimental restraints are used in molecular dynamics simulations in a time averaged fashion (MDtar), the experimental structural information does no longer need to be satisfied at each step of the simulation; instead, the whole trajectory must agree with the restraints. However, the resulting structural ensembles are large and not unique and it is not trivial to extract the essential dynamic features for a system. Here we demonstrate that large MDtar ensembles can be simplified substantially by reducing the number of members to just a few on the basis of adjusting the individual probabilities of the members with the PDQPRO program [N. B. Ulyanov et al. Biophysical Journal (1995), Vol. 68, p. 13]. This algorithm finds the global minimum for a search function that represents the best match of a given ensemble with the experimental dipolar interproton relaxation rates. We have applied this strategy to a 17-residue RNA hairpin, whose loop exhibited considerable flexibility evident from nmr data. This 17mer is a mimic of the T psi C-loop of tRNA, where nucleotide 54 is usually a ribosylthymidine. The methylation of U54, which is completely buried in transfer ribonucleic acid, is administered by tRNA (m5 U54)-methyltransferase (RUMT). Since the 17mer is a good substrate for RUMT, we previously concluded that the flexibility of the 17mer's loop is a key to how RUMT gains access to the methylation site [L. J. Yao et al. Journal of Biomolecular NMR (1996) Vol. 9. p. 229]. Application of the PDQPRO algorithm to the previously acquired MDtar trajectories allowed us to reduce the number of conformations from several hundred to one major and five or six minor conformations with individual populations from approximately 5% to approximately 50% without any deterioration in the match with the experimental data. The major conformation exhibits a continuation of A-form helicity through part of the loop, involving C60 and U59. In this and most other conformations the methylation site in U54 is no longer buried. PMID- 9754030 TI - Epidemiology of musculoskeletal disorders due to biomechanical overload. AB - The link between occupation and musculoskeletal disorders has been focused on in numerous research projects, ranging from those simply observing the different pathological findings reported among workers performing particular tasks, down to the latest studies actually quantifying the 'exposure' of workers to physical and psychosocial stimuli. For some disorders and certain tissues, it has been reported that specific types of work-related exposure are associated with the development of musculoskeletal pathologies, and that the relative risks for certain types of occupational exposure can be extremely high. This has been proven in relation to tendinitis of the shoulder and hand-wrist, carpal tunnel syndrome, as well as several localized aspecific musculoskeletal symptoms, such as pain. For other pathologies, the studies reported contradictory results. This is the case for lateral epicondylitis and cervical radiculopathy. PMID- 9754031 TI - An observational method for classifying exposure to repetitive movements of the upper limbs. AB - This article presents and discusses a model for describing and evaluating the principal risk factors characterizing occupational exposure: frequency and repetitiveness of movements; use of force; type of posture and movements; distribution of recovery periods; and presence of other influential (additional) factors. For each risk factor, the author proposes a method of practical detection in the field, as well as criteria for classifying and interpreting the results based on a critical review of the available literature on the subject. Numerous examples are supplied to better illustrate the concepts presented. The various factors considered are classified using numbers or indexes, so that they can be integrated into a concise exposure index. PMID- 9754032 TI - OCRA: a concise index for the assessment of exposure to repetitive movements of the upper limbs. AB - In the light of data and speculation contained in the literature, and based on procedures illustrated in a previous research project in which the author described and evaluated occupational risk factors associated with work-related musculoskeletal disorders of the upper limbs (WMSDs), this paper proposes a method for calculating a concise index of exposure to repetitive movements of the upper limbs. The proposal, which still has to be substantiated and validated by further studies and applications, is conceptually based on the procedure recommended by the NIOSH for calculating the Lifting Index in manual load handling activities. The concise exposure index (OCRA index) in this case is based on the relationship between the daily number of actions actually performed by the upper limbs in repetitive tasks, and the corresponding number of recommended actions. The latter are calculated on the basis of a constant (30 actions per minute), which represents the action frequency factor; it is valid- hypothetically--under so-called optimal conditions; the constant is diminished case by case (using appropriate factors) as a function of the presence and characteristics of the other risk factors (force, posture, additional elements, recovery periods). Although still experimental, the exposure index can be used to obtain an integrated and concise assessment of the various risk factors analysed and to classify occupational scenarios featuring significant and diversified exposure to such risk factors. PMID- 9754033 TI - Clinical trials among worker populations: a model for an anamnestic survey of upper limb pathologies and its practical application methods. AB - Following a brief review of the principal clinical characteristics of musculoskeletal disorders of the upper limbs, the authors propose a protocol for a structured anamnestic examination featuring a series of set questions. The anamnestic model is based on a detailed listing of the symptoms to be analysed, which are divided into four categories: pain, paraesthesia, symptoms attributable to hyposthenia, and neurovegetative disorders. Regarding pain and paraesthesia, the authors list the localization, pattern of onset, duration and number of episodes, irradiation and treatment. The patients can thus be classified as anamnestic cases based on the following criteria: presence of pain or paraesthesia during the last 12 months, with episodes lasting for at least 1 week or occurring at least once a month, with no previous acute trauma. For hyposthenia, the authors report on the conditions under which the disorder may develop. The neurovegetative disorders considered are modifications in colour of the fingers and reaction to exposure to low temperatures. The structure of the proposed anamnestic chart permits all findings to be easily encoded for subsequent storage in a dedicated database. The appendix contains an annotated facsimile of the anamnestic chart. PMID- 9754034 TI - Clinical trials among worker populations: the value and significance of anamnestic findings and clinical and instrumental tests for diagnosing work related musculoskeletal disorders of the upper limbs (WMSDs). AB - The authors discuss the value and significance of symptoms in WMSDs, in view of the fact that the anamnestic threshold proposed in epidemiological investigations cannot be used as clinical and diagnosing criteria. Some useful clinical procedures are suggested for cases where there is a suspicion of musculoskeletal disorders of the cervical spine and upper limbs, bearing in mind that they are to be applied within the framework of health surveillance programmes undertaken by health care practitioners who are not specialists in orthopaedics, physiatrics or neurology. The recommendations for instrumental tests and specialist referrals are also discussed for the various disorders. The authors also provide flow charts for the diagnostic procedures pertaining to WMSDs. The appendix shows a sample patient chart illustrating the proposed procedures; it also permits the findings to be encoded so that they can be stored in a dedicated database. The codes for diagnosing WMSDs are also reported for the same epidemiological purposes. PMID- 9754035 TI - The occurrence of musculoskeletal alterations in worker populations not exposed to repetitive tasks of the upper limbs. AB - A total of 749 workers (males: 139 aged between 15 and 35 years, and 171 aged > 35 years; females: 176 aged between 15 and 35 years, and 263 aged > 35 years) performing tasks not at risk for work-related musculoskeletal disorders of the upper limbs (WMSDs) underwent a clinical examination using a standardized method. The 'anamnestic cases' were defined on the basis of pain or paraesthesia present for at least 1 week during the previous 12 months, or appearing at least once a month, and not subsequent to acute trauma. The anamnestic cases among the males amounted to 4.4% (age 15-35 years) and 12.3% (age > 35 years); among the females, 4.6% (age 15-35 years) and 14.2% (age > 35 years). Of the 1498 limbs examined, the prevalent diseases reported were: suspect narrow chest syndrome: 0.3% among the males > 35 years, 0.6% among the females aged 15-35 years, 1% among the females > 35 years; scapulo-humeral periarthritis: 0.3% among the males aged > 35 years, 0.3% among the females aged 15-35 years, 1.3% among the females aged > 35 years; lateral epicondylitis: 0.3% among the males aged > 35 years, 0.2% among the females aged > 35 years; trapeziometacarpal arthrosis: 0.8% among the females aged > 35 years; wrist-hand tendinitis: 0.9% among the males aged > 35 years, 0.9% among the females aged 15-35 years; carpal tunnel syndrome: 2.5% among the females aged > 35 years. No disorders were detected outside the age ranges indicated. Several workers reported more than one disorder. The number of workers with at least one WMSD was: males 0% in the 15-35 years age range, 3.5% in the > 35 years age range; females 2.3% in the 15-35 years age range, 7.2% in the > 35 year age range; 3.9% of the total sample population. The prevalences were on average quite low, particularly among the older workers, hence the authors recommend that even minimal prevalences detected in particular work environments should not be underestimated. PMID- 9754036 TI - Application of the concise exposure index (OCRA) to tasks involving repetitive movements of the upper limbs in a variety of manufacturing industries: preliminary validations. AB - A summary of eight investigations is presented, which were carried out using standardized methods, for the purpose of quantifying exposure to tasks involving repetitive movements of the upper limbs, as well as quantifying the prevalence of work-related musculoskeletal disorders (WMSDs) of the upper limbs in groups of exposed workers. A total of 462 exposed workers were examined, and the study also took into account the data pertaining to a matched reference group comprising 749 workers not exposed to any specific occupational risk. Regarding the quantification of exposure to increased risk, use was made of a concise index (OCRA index), proposed by Occhipinti, in this issue. The data resulting from the eight investigations were used for the study of measurements and models of association among the exposure variables (mainly represented by the OCRA index), as well as the effect variables represented by the prevalence of the various WMSDs of the upper limbs taken both individually and jointly. Significant associations were reported between the OCRA index and an effect indicator represented by the prevalence of all the WSMDs of the upper limbs, calculated on the number of upper limbs at risk. When a logarithmic conversion of the relative exposure (OCRA) and injury indices was carried out, a simple linear regression model resulted that seems to provide a satisfactory predictive performance of the risk of WMSDs of the upper limbs, based on the exposure index. The study confirmed the efficacy of various other models designed to predict effects based on multiple linear regression functions, in which the independent variables are represented by both the OCRA exposure index and by parameters relative to the breakdown by gender and age of the groups of exposed workers. PMID- 9754037 TI - Criteria for the health surveillance of workers exposed to repetitive movements. AB - In the light of the experience and guidelines developed by other countries and of Italian legislative and operational conditions, the authors outline a strategy for a health surveillance programme for work-related musculoskeletal disorders of the upper limbs. In particular, the paper defines the various aims of the health surveillance programme and identifies significant relevant criteria for its implementation (i.e. existence of risks or effects). A screening schedule is presented based on subsequent investigations (first and second level surveillance); the authors discuss the principal methods used for processing the results of the health surveillance programme, in collective (i.e. statistical comparisons, planning of periodical investigations) and individual terms (job fitness judgements, reporting of suspected occupational diseases). PMID- 9754038 TI - Guidelines for designing jobs featuring repetitive tasks. AB - Preventive measures aimed at minimizing the occurrence of work-related musculoskeletal disorders of the upper limbs (WMSDs) associated with repetitive tasks can be divided into three categories: structural, organizational and educational. Whenever specific risk and injury assessments have shown the need for preventive action, this is most often implemented within the framework of a range of assorted measures. In particular, structural measures involve optimizing the layout of the work area and furnishings, and the 'ergonomic' properties of work tools and equipment. Such measures serve to alleviate the problems caused by the use of excessive force and awkward postures. The authors refer to the principles guiding such structural measures, in the light of the extensive literature that has been published on the subject. Organizational (or reorganizational) measures essentially relate to job design (i.e. distribution of tasks, speeds and pauses). They serve to alleviate problems connected with highly repetitive and frequent actions, excessively lengthy tasks and inadequate recovery periods. Very few relevant findings are available: the authors therefore illustrate in some detail a practical trial conducted in a major engineering firm. The objective was to lower to acceptable limits the frequency of certain repetitive tasks performed using the upper limbs. The trial made it possible to identify a suitable plan and schedule of measures taking into due consideration the impact of the plan on production levels (and costs). The fundamental principles guiding the adoption of specific educational and training programmes for the workers and their supervisors are presented and discussed. PMID- 9754039 TI - Criteria for the reintegration in the workforce of workers with musculoskeletal disorders of the upper limbs, based on preliminary practical experience. AB - This paper presents a preliminary study on the return to the workforce of employees with WMSDs of the upper limbs, and their reallocation to jobs with 'low exposure'. The study, which is still underway, involves a large engineering firm and includes some 100 workers affected by WMSDs. The trial involved: providing a definition of the criteria for characterizing 'accommodating' jobs (i.e. frequency of action < 20 actions/min; virtual absence of other risk factors such as force, awkward posture, inadequate pauses, etc.); effectively identifying jobs meeting such criteria (or jobs which, with minimal modifications, could be made suitable); classifying WMSD workers according to the type and severity of the disorder; matching WMSD workers with the jobs best suited to them; specific training for the workers and their supervisors; carrying out a follow-up of the return of WMSD workers to the workforce in organizational terms (i.e. the need for further modifications to equipment or procedures) and clinical terms (i.e. symptom patterns, acceptability of the condition). The preliminary results, 6-12 months after the start of the trial, are extremely encouraging, and show that when workers return to the workforce in jobs that fully meet defined criteria, a significant prevalence of 'improvements' are reported among the workers involved. The investigation will need to be extended but it already shows quite convincingly that it is possible for workers with what can be described as a 'reduced working capacity' to remain 'productive' (albeit in jobs featuring a lower exposure potential than the acceptable threshold for 'healthy' workers). PMID- 9754040 TI - Monoclonal antibody: high density culture of hybridoma cells and downstream processing for IgG recovery. AB - Getting higher yields of monoclonal antibody (MAb) is a problem in Hybridoma Technology which has two major bottlenecks--(a) poor yield of hybridized cells; and (b) low cellular productivity of MAb in culture. There are three ways of obtaining high MAb yield in vitro--(a) large scale culture of hybrid cells; (b) high density culture; and (c) enhancing individual cellular productivity in culture. Currently, focus is on correct synergistic combination of fortified nutrient media, bioreactor design and mode of operation. Maximisation of cell culture longevity, maintenance of high specific antibody secretion rates, nutrient supplementation, waste product minimization and control of environmental conditions are important parameters for improvement of large scale production of MAb. Though, MAb yield has enhanced rapidly over the decade, there is a growing concern for decrease in quality of MAb secreted. Further research is therefore necessary to take full advantage of MAb as a potential diagnostic agent for in vivo therapy. PMID- 9754041 TI - Calcium-dependent metabolic regulations in prokaryotes indicate conserved nature of calmodulin gene. AB - Role of free calcium and calcium binding protein calmodulin as signal molecule in cellular regulation is well established in eukaryotes. However, reports on Ca(2+) dependent processes and their inhibition by calcium and/or calmodulin antagonists indicate towards the presence of calmodulin in prokaryotes as well. The common evolutionary origin of pro- and eukaryotes and many examples of evolutionary conservation of structure and functions support the contention of such conservation of the role of Ca2+ and calmodulin. Eukaryotic calmodulin (CaM) contains four structurally and functionally similar Ca2+ domains named I, II, III and IV. Each Ca2+ binding loop consists of 12 amino acid residues with ligands arranged spatially to satisfy the octahedral symmetry of Ca2+ binding. Plant calmodulin differ from vertebrate ones in 13 to 14 amino acid positions of which nine occur at -COOH- terminal half. Differences between protozoan and mammalian CaM also occur mostly in the same half. Isolation and characterization, although to a little extent, of CaM-like proteins from bacteria and cyanobacteria and their comparison with CaMs from diverse origin suggest high degree of conservation. Non-bulky amino acids like glycine, alanine and serine with low specific rotation are present in greater number in the primitive form of calmodulin and have been significantly reduced in highly evolved form of calmodulin, suggesting that their requirement was insignificant and were eliminated from EF hand structure during evolution. However, amino acids like glutamate/glutamine and aspartate/asparagine were highly conserved and did not show any major change in their frequency since their positions are too significant in calcium binding domain. While the number of positively charged amino acids like arginine and leucine was increased, histidine containing weakly ionized group and having a significant buffering capacity was reduced to a major extent, further suggesting that the acidic nature of calmodulin protein has been maintained during evolution. Thus it is now clear that the entire superfamily of Ca2+ binding proteins have arisen from a common genetic ancestry. Two successive tandem duplications of gene encoding a single domain containing protein of 30-40 residues gave rise to a four domain molecule from which this family was then derived. PMID- 9754042 TI - Effects of hypothyroidism and hyperthyroidism on serum glycoproteins in experimental rats. AB - Hypothyroidism and hyperthyroidism were induced in adult female Sprague Dawley rats by feeding 0.05% propylthiouracil and 0.0012% L-thyroxine in drinking water. Contents of protein-bound hexose, fucose, hexosamine and sialic acid in serum were estimated in experimental rats. The results showed an increase in the protein bound hexose, fucose, hexosamine and sialic acid levels in hypothyroid rats and decrease in hyperthyroid rats. The increased content of serum glycoproteins in hypothyroid rats might possibly have come from disruptions of membranes. Depleted glycoproteins in hyperthyroid animals might be due to increase in depolymerisations of glycoproteins and accelerated secretion of glycoproteins with urine. PMID- 9754043 TI - Effects of calcium on phthalocyanine sensitized photohemolysis. AB - Influence of divalent Ca2+ ions in sulphonated chlorophthalocyanine (ClAlPcS2) induced photohemolysis in rabbit red blood cells has been investigated. Loading of excess Ca2+ to ClAlPcS2 sensitized RBC suspension enhanced hemolysis in a concentration dependent manner. Protein kinase C (PKC) inhibitor chlorpromazine and cation chelator EDTA reduced Ca2+ enhanced photohemolysis. Photohemolysis increased with calcium channel blockers diltiazem and nifidipine. Enhancement in photohemolysis by Ca2+ ions can arise either by its ability to liberate Fe2+ bound to phosphatidylserine or by stimulation of PKC. Inability of calcium channel blockers to prevent Ca2+ influx following photodynamic treatment of red blood cells suggest nonspecific leakage rather than involvement of Ca2+ channels in erythrocytes. PMID- 9754044 TI - Effect of medroxy progesterone acetate and testosterone enanthate on vas deferens of rats. AB - Effects of medroxy progesterone acetate (MPA; 5 mg/kg) and MPA + testosterone enanthate (TE) (3 mg + 2 mg/kg) were investigated in vas deferens and on fertility (along with reversibility study) for 60 days through histopathology, morphometric and certain biochemical parameters such as total proteins, sialic acid, ATPase, SDH and testosterone. The study revealed for altered histopathology and contratile pattern of vas deferens resulting in reduced fertility. The study also indicated androgen antagonistic effect. These effects were found to be reversible 4 and 3 months after withdrawal of MPA and MPA + TE injections respectively. Thus, both types generated functional sterility in the rat, but MPA affected histophysiology of vasal tissue. PMID- 9754045 TI - Statistical significance of AgNOR counts in FNAC smears and corresponding histopathological sections. AB - The object of the present study was to determine the statistical significance of AgNOR counts in fine needle aspiration cytology (FNAC) smears and corresponding paraffin sections by using an one step silver colloidal staining method. Ninety five cases (31 benign and 64 malignant) were taken and a correlation between smears and sections was studied by two different observers. The total number of mean (SD) AgNOR counts was significantly higher in FNAC smears in benign (3.081 +/- .753) and malignant (7.101 +/- 1.544) neoplasms in comparison to paraffin sections in the same group of cases. FNAC smears had a cut off point 4 with proliferation index 1% in benign tumors and 97.5% in malignant tumors. Sections had an overlapping of AgNOR counts (5-7) with proliferation index 3.8 and 82% for benign and malignant groups respectively. The difference in the coefficient of variations was 3% in benign group and 2% in malignant group in FNAC smears while sections had 6 and 8% difference in the coefficient of variations. Smears present a superior staining and accurate number of AgNOR dots in nucleus as compared to paraffin sections. Therefore the results suggest that AgNOR technique can be successfully used in FNAC smears in comparison to paraffin sections to differentiate benign and malignant tumors in routine laboratory diagnosis. PMID- 9754046 TI - Immunization of rabbits against Hyalomma anatolicum anatolicum using homogenates from unfed immature ticks. AB - Antigens were prepared from unfed larvae and nymphs of H. a. anatolicum as homogenised antigens (HLAg and HNAg, respectively). Five rabbits each were inoculated, s.c. with 8.56 mg HLAg and 9.34 mg HNAg in 3 divided doses. Following immunisation rabbits developed significant level of protective immunity to infestation with adults of this species. Significant reduction in engorged percentage and weights of engorged females and egg masses were observed in females fed on immunized rabbits, compared to that of female ticks fed on control rabbits. The engorgement period was also increased significantly. However, conversion efficiency indices remained unaffected. Larval antigen immunized rabbits showed significant antibody level from 28-126 days while with HNAg elevated antibody levels were recorded up to 112 days. Further, the rabbits immunized with HLAg had elevated level of antibodies against HLAg, HNAg, and adult antigen in ELISA. But HNAg immunized rabbits had lower levels of antibodies against HLAg and HAAg as compared to values recorded against HNAg. Anti-HLAg and anti-HNAg sera recognised common antigenic bands of 97.4, 85, 66, 47.3, 42 and 31 kDa in homogenates of larvae, nymphs and adults. PMID- 9754047 TI - Effect of leaf extract from Clerodendron colebrookianum on CNS function in mice. AB - Effect of acute multiple (20, 40 and 80 mg/kg body wt, i.p.) doses of C. colebrookianum leaf extract on behaviour, convulsion, analgesia and sedative hypnosis was studied in mice. A marginal reduction of awareness and motor activity was observed in low (20 mg) and moderate (40 mg) dose level of extract. However, 80 mg dose caused marked inhibition of awareness and motor activity. Grip strength and stereotypy was observed in all the dose levels. The extract alone did not show loss of righting reflex but it prolonged the effect of meprobamate, diazepam, chlorpromazine and pentobarbitone significantly in a dose dependent manner. The extract neither produced analgesia alone nor it altered analgesic effect of morphine and pethidine. Pretreatment of the extract caused significant protection of strychnine and leptazol induced convulsion and mortality. The results suggest a mild (or dose dependent) CNS depressant action of leaf extract of C. colebrookianum in mice. PMID- 9754048 TI - Hepatoprotective effect of a protein isolated from Cajanus indicus (Spreng) on carbon tetrachloride induced hepatotoxicity in mice. AB - Treatment with hepatotoxin namely carbon tetrachloride (CCl4) (0.1 ml/100 g of body weight; twice a week) induced acute hepatic necrosis in Swiss albino mice (male; body weight 30 g +/- 2), with significant alteration in the activities of glutamic oxaloacetic transaminase (GOT); glutamic pyruvic transaminase (GPT); alkaline phosphatase (AP) and serum bilirubin. Administration of a protein fraction isolated from the leaves of C. indicus counteracted the action of CCl4 on transaminase, phosphatase showing hepatoprotection. Daily treatment with a purified protein fraction 'X' from the above plant (0.5 mouse ml i.p; 50-60 micrograms/ml) for a period of 7, 14, 21 days respectively showed decreased activities of serum transaminases alkaline phosphatase and decreased levels of serum bilirubin. These findings were further confirmed by histopathological study of liver. PMID- 9754049 TI - Effect of GABA and baclofen on gastric mucosal protective factors. AB - GABA and baclofen (BAC), a GABA-mimetic agent, were investigated for antiulcerogenic activity. Orally administered GABA (100 mg/kg) and BAC (10 mg/kg) showed significant ulcer protection when given either alone for one day or for 4 days, or when given together with aspirin (ASP; 200 mg/kg x 3 days) in their 4 days treatment time in pylorus-ligated rats. Both the drugs showed a tendency to increase acid and decrease peptic output, and increased gastric mucus secretion in terms of total carbohydrate to protein ratio (TC:P) in both the above treatment groups. ASP tended to decrease acid and increase peptic output and significantly decreased TC:P ratio. Both GABA and BAC tended to reverse aspirin induced effects, though they had little per se effect on TC:P ratio of gastric mucosal glycoproteins except an increase in sialic acid content both after one day or four days treatment. No, per se, effect on cell shedding (DNA and protein content of gastric juice) or cell proliferation (DNA/mg protein) was noted with GABA or BAC but the enhanced cell shedding induced by ASP was attenuated by them. ASP was found to enhance cell proliferation. However, neither of drug showed any effect on cell proliferation when given either alone or in combination with ASP. The antiulcerogenic effect of GABA and BAC may be due to their predominant effects on mucosal defensive factors like enhanced mucin secretion and decreased cell shedding or mucosal damage. PMID- 9754051 TI - Correlation between human papillomavirus DNA detection in maternal cervical smears and buccal swabs of infants. AB - Acquisition of human papillomavirus (HPV) infection in buccal mucosa by the infants at birth has been investigated. Presence of HPV DNA was evaluated in cervical smears of 30 pregnant women before delivery and in buccal swabs of the corresponding 31 infants (1 set of twins). HPV DNA was detected among the 40% of women, 16% of infants an the detection was concordant in 23 mother-infant pairs. HPV DNA was demonstrated in buccal mucosal cells of 41.6% of the infants born to HPV positive mothers. Maternal-infant transmission was highest for HPV 6/11. Assessment of the risk of developing HPV related oral lesions in children at later life owing to acquisition of HPV during perinatal period may help in determining a strategy to combat the disease. PMID- 9754050 TI - Pharmacological actions of Abies pindrow Royle leaf. AB - Petroleum ether (PE), benzene (BE), chloroform (CE), acetone (AE) and ethanolic (EE) extracts (50-200 or 200 mg/kg, i.p. or 200 mg/kg, p.o.) of dried Abies pindrow leaves, given 30-45 min before showed significant anti-inflammatory (both against acute and sub-acute models), analgesic, barbiturate hypnosis potentiation and anti-ulcerogenic acitivities in rats. All the extracts except EE decreased swim stress immobility in mice indicating some degree of anti-depressant activity. Only PE exhibited hypotension in dogs blocked by atropine. Chemically, extracts showed the presence of glycosides, steroids, terpenoids and flavonoids. They had no anti-bacterial effect. However, toxicity studies indicated that the extracts had an extended safety index. The investigations are consonant with some of the uses of this plant in Ayurveda. PMID- 9754052 TI - Effect of selenium deficiency and its supplementation on DTH response, antibody forming cells and antibody titre. AB - Selenium levels and the activity of Selenoenzyme glutathione peroxidase were measure in whole blood in order to assess the selenium status. Delayed type of hypersensitivity (DTH) reaction was suppressed significantly in selenium deficient rats indicating the decrease in cellular immunity. The B cell function was impaired in selenium deficient rats as evident from decrease in the number of plaque forming cells and antibody titre. Selenium supplementation for 30 days recovered the DTH response and B cell function to a marked extent. PMID- 9754053 TI - Localization of hup determinants in Rhizobium sp. sesbania. AB - Rhizobium sp. sesbania strain SRS 78, a Hup wild type carrying three plasmids was mutagenised using Tn5-sac BR cassette and clones carrying Tn5 insertion in plasmids were identified on the basis of transfer frequency of Tn5-sac BR labelled plasmids from Rhizobium to A. tumefaciens. Neomycin sensitive (Nms) clones were isolated from clones containing Tn5 labelled plasmids by plating on sucrose (5%). Nms clones did not show loss of hup activity. Preliminary results on curing studies indicated that hup genes are located on chromosome. These results were later confirmed by southern hybridization using plasmid pKHT30 containing hup genes of Azotobacter chroococcum. None of the three plasmids showed hybridization with hup gene probe while chromosomal DNA left in the wells showed positive signal. PMID- 9754054 TI - Is gene therapy for aging possible? AB - Throughout human history, search for means to prevent or slow down aging has followed three main lines--(1) removing waste products and cleansing impurities; (2) using products of plants and animals as medicine; and (3) compensating for decrease in various hormones, vitamins and other chemicals in the body. Even in modern times, immense popularity of various spas and water therapies is an example of the first type of anti-aging approach for which there are no real scientific basis. Some preliminary support from laboratory and/or clinical tests is available for various herbal and other medicinal plant products, such as ginseng, ginkgo biloba and garlic, as nutritional supplements. Replacement therapy, especially hormonal replacement therapy as an anti-aging treatment has been used and misused for quite some time. PMID- 9754055 TI - Bone morphogenetic proteins--novel regulators of bone formation. AB - Bone morphogenetic proteins (BMPs), hydrophobic, non-species specific glycoproteins, belong to the expanding transforming growth factor-beta (TGF-beta) superfamily. BMP has pleiotropic function that range from extraskeletal and skeletal organogenesis to bone generation and regeneration. It induces de novo bone formation in post fetal life through the process of direct (intra membranous) and endochondral ossification and their response is dose dependent. Through recombinant gene technology, BMP is available at least in ten forms for basic research and clinical trials. Amino acid sequences and physical properties of BMP family members have been identified. BMP research extends to the fields of developmental biology, genetics and evolution. PMID- 9754056 TI - Loss of a putative novel growth inhibitory apoptotic 14 kD polypeptide during progression of rat liver carcinogenesis. AB - Carcinogenesis is a multistep process involving different stages. However, the biological and biochemical factors responsible for the stepwise transition of cells from one stage to the other remains as important enigmas even today. We have recently isolated a putative novel growth inhibitory apoptotic 14 kD polypeptide from normal rat liver. In order to understand the possible functional relationship between 14 kD polypeptide and liver carcinogenesis, the sequential expression of this polypeptide as a function of tumor progression was studied in the rat liver using diethylnitrosamine (DEN) as a carcinogen. Immunoperoxidase and immunoblotting experiments using polyclonal rabbit antisera revealed a gradual reduction in the levels of this polypeptide with tumor progression. No reduction in the levels of this polypeptide was observed in regenerating rat liver after partial hepatectomy. The findings suggest that the loss or reduction of 14 kD polypeptide is linked selectively to abnormal cell proliferation and appears to be a biologically relevant risk factor for the progression of hepatocarcinogenesis in rats. PMID- 9754057 TI - Changes in membrane fluidity induced by binding of immunoglobulins to sperm Fc receptors. AB - Receptors for Fc region of immunoglobulins (FcR) are reported to be present on spermatozoa, and also detected in seminal plasma, however their function is still not known. Since various changes in sperm membrane architecture during maturation and passage through female genital tract are reported, experiments were conducted to study the membrane fluidity changes in sperm subsequent to ligation of surface FcR with immunoglobulin and its derivatives. This paper reports that interaction with IgG restricts rotational mobility of cell surface proteins and membrane lipids as studied by EPR spectroscopy using spin probes. Decrease in fluidity was much more pronounced in the presence of aggregated IgG due to crosslinking of sperm FcR by aggregated IgG. PMID- 9754058 TI - Central nervous system depressant activities of melatonin in rats and mice. AB - Melatonin (10 and 20 mg/kg) potentiated pentobarbitone (45 mg/kg) induced hypnosis; it (50 and 100 mg/kg) decreased both total and ambulatory activity; produced (10, 25 and 50 mg/kg) hypothermia and possessed (10-400 mg/kg) significant analgesic property both in tail-flick and acetic acid-induced writhing tests. Melatonin in low dose (10 mg/kg) significantly reduced the forced swimming induced immobility period per se and also reserpine induced immobility. While the antinociceptive effect of melatonin was sensitive to reversal by naloxone (2 mg/kg), other CNS depressant profile resembled the effects of benzodiazepines. PMID- 9754059 TI - Toxic effect of systemic administration of low doses of the plasticizer di-(2 ethyl hexyl) phthalate [DEHP] in rats. AB - DEHP [di-(2 ethyl hexyl) phthalate], a widely used plasticizer in blood storage bags, leaches out in appreciable amounts into blood (about 10 mg/100 ml) resulting in exposure of recipients of blood transfusion to this compound. Various reports indicate the toxicity of DEHP, particularly in liver and reproductive organs but all these studies used large doses (up to 2 g or more/Kg body weight) and oral route of administration which are not relevant to the intravenous administration during blood transfusion or the low amounts present in blood. We have studied changes in the activity of some important enzymes-gamma GT, ALT, CPK, LDH, alkaline phosphatase, acid phosphatase, beta-glucuronidase and few other parameters like vitamin E, glutathione, serum albumin etc in rats administered low doses of DEHP (corresponding to transfusion of 2, 4, 6 and 10 units of blood). Histopathology of the organs has also been carried out. The results obtained indicate no serious toxic effects for DEHP at the level present in blood stored in DEHP plasticized blood bags as evidenced by the lack of any significant alteration in most of the biochemical parameters studied. Even in those cases where there was alteration (for e.g., decrease in the level of vitamin E) 24 hr after administration of DEHP, it returned to near normal level with in 72 hr to 7 days. No histopathological changes were observed in any of the organs at these levels of DEHP. It is concluded that DEHP did not cause any serious toxic effect even at doses corresponding to transfusion of several units of blood in a recipient. PMID- 9754060 TI - A comparative evaluation of immunotoxicity of malathion after subchronic exposure in experimental animals. AB - The effects of sub-chronic doses of malathion exposure on humoral and cell mediated immune (CMI) responses were studied in male albino mice, rats and rabbits using sheep red blood cells (SRBC), tetanus toxoid and ovalbumin as antigens. The humoral immune response was assessed by estimating serum immunoglobulin (IgM and IgG) concentrations, antibody titre against antigens and splenic-plaque forming cells (PFC). The CMI response was studied by using the leucocyte migration inhibition (LMI) and macrophage migration inhibition (MMI) tests. In general there were (a) attenuation in antigen induced antibody response, (b) suppression of PFC, and (c) marked inhibition of LMI and MMI factors. Sub-chronic malathion exposure induced differential degrees of humoral and CMI suppression in these experimental animals. However, both cellular and humoral immune responses were decreased in a dose-time dependent pattern and a consistent trend was observed. The threshold level of the malathion for inducing immune suppression depends on the animal species, type of antigen used, and the method of immunological assay. In view of the widespread use of malathion a comparative assessment of immune responses using different experimental animals and antigens is an important aspect of its safety evaluation. PMID- 9754061 TI - Role of iron and copper during benzene toxicity. AB - Role of transition metal ions in expression of benzene toxicity has been suggested. Intraperitoneal administration of benzene to female albino rats daily for 10 days resulted in accumulation of iron in liver nuclei, without any change in copper content. Incubation of hydroquinone (HQ), one of the principal metabolites of benzene with rat liver nuclei resulted in formation of thiobarbituric acid reactive products (TBAR). However, presence of bathocuproine, a copper chelator and EDTA, an iron chelator caused significant inhibition of TBAR release. Thus, the present study revealed that iron accumulation and involvement of copper in nuclear damage induced by HQ. PMID- 9754062 TI - Pharyngeal colonisation, ADNB response and development of cardiac lesions in mice following intranasal inoculation with group A beta haemolytic streptococcus M type 18. AB - Swiss albino mice from a randomly bred colony were inoculated intranasally with 1.6, 2 or 2.4 x 10(7) colony forming units of a mid-logarithmic phase culture of group A beta haemolytic streptococcus M type 18 for 3 days, 6 days or once a week respectively for three weeks. Pharyngeal colonisation could be observed in 67 (59.8%) of 112 mice on 4th day after inoculation and 14 (38%) of the 37 mice on 21st day. Out of 27 mice tested for determination of antibodies to deoxyribonuclease B at regular intervals for 98 days, 15 (55.5%) showed responses, with maximum titers varying from 50 units to 4800 units in individual mice. Histopathological evidence for cardiac lesions were seen in five (3.03%) of the 165 animals studied. These included one case of severe endocarditis, two cases of endocarditis with valvular lesions and one case with non-specific lymphocyte infiltration in the heart. One other animal showed subendocardial oval nodular aggregates. Although the cellular nature of these lesions were not determined, this study shows that Swiss albino mice can serve as suitable animal models to study experimental streptococcal infections. However these are preliminary observations and are to be confirmed and revalidated by further controlled experiments. PMID- 9754063 TI - Isolation and characterization of Salmonella enterica Weltevreden cytotoxin. AB - Cytotoxin of Salmonella enterica subspecies enterica serovar Weltevreden (BM 1643), isolated from buffalo meat, was purified and characterized physicochemically and immunologically. Cell-free culture supernatant (CFCS) of the organism showing marked cytotoxicity to Vero cells and least enterotoxicity to rabbit ligated ileal loop (RLIL) model, was salt precipitated with ammonium sulphate (60% saturation level) and dialysed. Precipitated dialysed preparation (60% PDP) when filtered through Sephadex G-100 column yielded two peaks, of which second peak (SG-100 SP) contained the cytotoxic activity. Upon filtration of SG 100 SP through SG-200 column, three peaks were obtained. Second peak (SG-200 SP), which was cytotoxic, yielded a single protein band of approximately 60-70 kDa in polyacrylamide gel electrophoresis and 3 protein bands of lower, molecular weight (13.5-56 kDa) in SDS-PAGE analysis. Cytotoxic preparation was maximally active at pH 7 to 8. On heating above 60 degrees C, cytotoxicity decreased gradually with insignificant activity left after treatment at 121 degrees C (15 min). Cytotoxin was inactivated by treatment with trypsin and protease but not by papain or lipase enzymes. It was immunogenic in rabbit and antiserum neutralized the cytotoxicity of cytotoxic preparations of homologous as well as heterologous Salmonella serovars. PMID- 9754064 TI - Hyperglycemic effect of low protein cassava diet. AB - Hyperglycemic effect of cassava diet in presence of varying amounts of protein has been carried out. The rats fed a low protein high cyanide diet showed an increase in the blood glucose and a decrease in the liver glycogen. The activity of glycogen phosphorylase, glucose 6-phosphatase and phosphoglucomutase showed higher levels in the liver of low protein high cyanide group compared to the control group. Also, the activity of hexokinase, and isocitrate dehydrogenase activity in the liver of high cyanide low protein were significantly low. The results suggests that cassava diet with the low protein can induce hyperglycemia. PMID- 9754065 TI - Effect of calcium-channel blocking drugs on focal ventricular arrhythmias induced by sub-epicardial epinephrine infusion in cats. AB - Sub-epicardial infusion of epinephrine (EP) in the dose of 3 x 10(-3) M in 2.5 x 10(-3) M CaCl2-0.9% NaCl (calcium-saline vehicle) at the rate of 10 microliters in the right ventricular myocardium of mongrel cats weighing between 2.8 and 3.3 kg, produced uniform, reversible and reproducible focal ventricular arrhythmia (VA) of varrying intensity and duration. Infusion of two calcium-channel blocking agents, verapamil (VP) and nifedipine (ND) at the site of arrhythmogenesis, in equimolar concentration of 3 x 10(-3) M alongwith EP in the same vehicle reduced the incidence, duration, peak and mean frequencies of arrhythmias while, the latent period of onset of arrhythmias increased significantly. Verapamil in equimolar concentration of 3 x 10(-3) M was found to be more effective than nifedipine in antagonizing EP-induced ventricular arrhythmias. PMID- 9754066 TI - Effect of rhinax on bacterial lipopolysaccharide induced endotoxemia in rats. AB - Administration of lipopolysaccharide (LPS) at 3 mg/kg, i.p. in rats resulted in reduced food intake, febrile hyperthermia, decreased body weight and reduced muscle performance in treadmill tests. It also induced some biochemical changes like increased serum levels of transaminases, acid phosphatase, pseudocholinesterase, free fatty acids and decreased blood glucose and liver glycogen levels. Rhinax (RHX), a herbal formulation, at 160 mg/kg, p.o. improved muscle performance but had no effect on the elevated temperature or the reduced body weight of rats weakened by LPS. It also normalised various biochemical alterations induced by LPS. The results of these studies indicate efficacy of RHX as an antifatigue agent to improve muscular performance. PMID- 9754067 TI - Possible significance of anogenital licking by female rat during precopulatory behavioural patterns. AB - Possible significance of licking of anogenital region by female rat in the initial steps of precopulatory behaviour was studied in short bouts of 3 min each. The order of conspicuous events was--(1) exploratory behaviour, (2) partners pursuing each other, (3) head-to-tail orientation and mutual licking of anogenital regions; particularly when females were in oestrous and metoestrous stages and (4) obvious avoidance and even aggressive behaviour by female during dioestrous stage. PMID- 9754068 TI - Biodegradation of dimethylterephthalate by Comamonas acidovorans D-4. AB - Comamonas acidovorans D-4, capable of utilizing dimethylterephthalate (DMT) as the sole carbon source, was isolated from the activated sludge of petrochemical wastewater treatment plant. Almost complete utilization of as high as 0.5% (w/v) DMT was observed in 72 hr. Growth kinetics followed a parallel relation between the growth, DMT utilization and cell associated esterase activity. A cell free broth of DMT grown cells showed an extracellular esterase activity. During the DMT degradation an extracellular accumulation of terephthalic acid was found. Although, C. acidovorans grew on a number of phthalate esters and phthalic acids as the sole carbon source, growth was significantly high on phthalic acids. The potential of this organism in petrochemical pollution abatement is discussed. PMID- 9754069 TI - American Society of Critical Care Anesthesiologists, 11th annual meeting. Orlando, Florida, USA. October 16, 1998. Abstracts. PMID- 9754070 TI - 27th Annual meeting of the International Society for Experimental Hematology. Vancouver, Canada. August 1-5, 1998. Abstracts. PMID- 9754071 TI - American Society for Therapeutic Radiology and Oncology 40th annual meeting. Phoenix, Arizona, USA. October 25-29, 1998. Abstracts. PMID- 9754072 TI - International Society of Neuroimmunology 5th International Congress. Montreal, Canada. 23-27 August 1998. Abstracts. PMID- 9754073 TI - The American Dietetic Association. 81st annual meeting and exhibition. Kansas City, Missouri, USA. October 19-22, 1998. Abstracts. PMID- 9754075 TI - [Of other AIDS-related cancers]. PMID- 9754076 TI - [Cancer of the breast: the return of lymphoscintigraphy?]. AB - Lymphoscintigraphy, after arousing great hope in the past in the field of breast cancer, has now been abandoned. The inability of this examination to predict the metastatic status of the nodes, and progress in therapeutic concepts have led to abandoning this technique. However, certain problems encountered by regional irradiation programmes and the work concerning sentinel node detection may bring this technique back into the spotlight. Lymphoscintigraphy may make it possible to adopt an individual approach, case by case, of the lymphatic drainage basins in breast tumors, thus enabling certain patients to benefit from regional irradiation when it would not have been traditionally recommended for this irradiation. Another aspect concerns the problem of the volumes irradiated. Work carried out with lymphoscintigraphy has enabled internal mammary chain nodes to be precisely located. Theses studies show the necessity of adapting the irradiation field to each individual case, but the clinical impact is limited, in the end, by the low recurrence rate in the internal mammary chain area. However, the new techniques of computer merging of scintigraphic and scanner images could enable the spatial position of the nodes in the upper axillary and supraclavicular regions to be determined. This would have, a priori, much wider clinical impact. Lymphoscintigraphic detection of the sentinel node is another field of major interest, but this technique is in competition with staining techniques. This procedure leads to a large reduction in morbidity of axillary surgery in 70% of patients. The use of techniques for detecting micrometastases in the sentinel node opens prospects in terms of prognosis. The qualities of differents radiotracers and different injection sites possible are also discussed. PMID- 9754077 TI - [The Fas/Fas-ligand system: implications in the antitumor immune response and in the activity of cytotoxic agents]. AB - Interaction of Fas-ligand (Fas-L) with the extracytoplasmic domain of the Fas receptor can induce Fas trimerization and activation of the apoptotic cell death process. Several molecular pathways that lead to apoptosis and some of their regulatory mechanisms have been identified. Fas-related membrane receptors that contain a death domain in their intracytoplasmic domain have been identified. They constitute a death receptor family (DR1 to DR5) whose first member is the TNFR1 receptor for TNF alpha. The Fas/Fas-L system plays a role in the cytotoxic activity of immune cells and the regulation of immune response amplitude. This system could be involved in the immune response to tumor cells and the cytotoxic activity of drugs and radiations. The expression of Fas-L on the plasma membrane of numerous tumor cells allow them, in vitro, to kill Fas-expressing immune cells. This observations has suggested that tumor cells used Fas-L to induce a specific immune tolerance. However, in vivo, Fas-L expression rather induces tumor cell rejection. The quantity of Fas-L expressed on tumor cells could determine whether tumor cells are tolerated or rejected. Cytotoxic drugs and radiations modulate Fas and Fas-L expression on tumor cells. The role of Fas/Fas L interactions in the cytotoxicity of these agents remains poorly defined. It has been clearly shown, however, that low doses of cytotoxic drugs increase Fas expression on tumor cells, thereby improving their elimination by immune cells. Drug-induced modulation of Fas expression could provide new therapeutic strategies combining chemotherapy with immunotherapy. PMID- 9754078 TI - [Standards, Options, and Recommendations for the management of brief neutropenias. Federation Nationale des Centres de Lutte Contre le Cancer]. AB - CONTEXT: The "Standards, Options and Recommendations" (SOR), initiated in 1993, is a collaborative project between the Federation of the French Cancer Centres (FNCLCC), the 20 French Cancer Centres and specialists from French Public Universities, General Hospitals and Private Clinics. The main objective is the development of clinical practice guidelines to improve the quality of health care and outcomes for cancer patients. The methodology is based on literature review and critical appraisal by a multidisciplinary experts group, with feedback from specialists in cancer care delivery. OBJECTIVE: To develop a clinical practice guideline for the management of neutropenic cancer patients (excluding prolonged neutropenia). METHODS: Data have been identified by literature search using Medline and Current Contents (up to February 1997) and personal reference lists. The main end points considered were mortality, morbidity, risk factors, fever, source of infection, microbiological documentation, incidence and length of hospital stays, quality of life, efficacy of treatment, safety and costs. Once the guideline was defined, the document was submitted to 48 reviewers for peer review and to the medical committees of the 20 French Cancer Centres for review and agreement. RESULTS: The key recommendations are: 1) before receiving cytotoxic chemotherapy, patients must be informed of potential risks and precautions to observe; 2) non-febrile neutropenic patients can be followed at home (except specific context); antibiotic prophylaxis is not recommended; 3) initial empirical antibiotic therapy for febrile patients is mandatory, whether associated beta-lactam and aminoglycoside, or monotherapy with a broad-spectrum beta-lactam (except in case of septic shock or pneumopathy). A glycopeptide can be added in case of overt catheter-related or cutaneous infection, in case of microbiologically documented infection with a oxacillin-resistant Gram positive bacteria, or in case of persistent fever in a clinically deteriorating patient; 4) at the present time, there is insufficient evidence to recommend the management of febrile neutropenic patients at home. We recommend participation in studies to identify predicting factors of low-risk patients and to assess the feasibility and safety of early discharge and home therapy. PMID- 9754079 TI - [Contralateral axillary lymph node metastasis of cancer of the breast]. AB - Contralateral axillary metastases (CAM) of breast cancer are uncommon, they state some diagnostics and therapeutics problems. From February 1993 until June 1996, 6 cases of CAM were detected in 123 women having a breast's cancer (4.9%). The diagnostic of CAM, was retained after a normal clinical and mammographic examination of ipsilateral breast, and a anatomopathological proof confirming the carcinomatous infiltration of axillary ganglions. These metastases were synchronous for 3 patients and metachronous for the 3 others with an appearance's mean delay of 12 months. The 6 patients were treated for a breast's cancer locally advanced and a tumour with internal or central seat in 5 cases. The CAM were associated to others metastatic areas in 2 patients. Five patients have had a locoregional treatment of CAM (curage and/or axillary radiotherapy), associated to chemotherapy. The 6th patient, is multimetastatic, had refused the treatment. Only 1 patient has developed a tumor of ipsilateral breast after a mean delay of 7 months, others metastatic localizations are appeared in 2 patients after a mean delay of 8 months. The CAM are considered a scare entity with physiopathological mechanism not yet well elucidated. They are perhaps, the prerogative of advanced tumors and the often before metastatic spreading. A problem remains to be raised: is it a question of a real CAM, or an expression of a ipsilateral breast's occult carcinomatous? PMID- 9754080 TI - [Importance of a multidisciplinary approach to metastatic cancer of the rectum]. AB - Management of rectal cancers with synchronous metastasis is difficult. We evaluated in 23 patients a combination of pelvic radiotherapy at the dose of 45 Gy in 5 weeks and 25 fractions with chemotherapy by 5-fluorouracil (350 mg/m2/day) and folinic acid (20 mg/m2/day) for 5 days at the time of the first and the fifth week of the irradiation. Surgery was indicated firstly in cases of stricture or secondarily for resection of the primary location and, when possible, of the metastasis. General state of health of the patients improved in 35%, symptomatology in 86% and comfort in 72% of the cases. Response rates for the primary tumor were 41% of partial response and 50% of stable disease. For the metastatic lesions, they were 9% and 59% respectively. Sixty-one per cent of patients were secondarily operated with resection of the primary tumor in 12 cases and of hepatic metastases in 2 cases. The median survival and the median survival without progression were respectively 13 and 9 months. Radiochemotherapy combination as the first treatment was beneficial in 4/5 of the patients presenting a rectal cancer with synchronous metastasis and allowed us to select those that would secondarily benefit from a surgical resection. PMID- 9754081 TI - [Endometrial evaluation prior to tamoxifen: preliminary results of a prospective study]. AB - The objective of this study was to try to identify, by pretreatment screening, a group of patients at higher risk of developing endometrial carcinoma on tamoxifen. Between January 1993 and January 1997, 360 postmenopausal patients with breast cancer were enrolled in this prospective study. Basal screening included gynaecologic examination with a Papanicolaou smear and endovaginal sonography. In the case of an abnormal ultrasound (endometrial thickness greater than 4 mm), an outpatient hysteroscopy with an endometrial biopsy was carried out. These examinations were repeated annually. By means of this preliminary evaluation, two groups of patients were identified: patients without initial lesions (group I) and patients with initial endometrial lesions (group II). These two groups of patients were followed up separately exactly in the same way. Endometrial lesions taken into account were: adenocarcinomas (in situ and invasive), polyps with or without atypia, myomas and adenomyotic lesions with irregular mucosa. After 3 years and after 4 years of follow-up, the percentage of atypical lesions was significantly higher in the group with initial lesions than in the group without initial lesions. This study suggests that a group of high risk patients more sensitive to the carcinogenic effect of tamoxifen can be identified by pretreatment evaluation. PMID- 9754082 TI - [Chemoprevention of cancer of the breast. Position statement on July 3, 1998 of the Fedration Nationale des Centres de Lutte Contre le Cancer (FNCLCC)]. PMID- 9754083 TI - Standardizing the classification and description of follicular unit transplantation and mini-micrografting techniques. The American Society for Dermatologic Surgery, Inc. AB - Previous attempts at classifying small graft transplants have focused mainly upon graft size and have not taken into consideration other technical factors involved in graft production that may influence the outcome of the surgery. The proposed classification attempts to consider these factors by including various technical aspects of harvesting, dissection, and placement, all of which impact the quality and quantity of the small grafts used in the procedure. By standardizing the nomenclature, as well as the description of the other factors involved in the surgery, communication between physicians and patients may be facilitated. In addition, different procedures may be more accurately studied and compared. PMID- 9754084 TI - Is there a rationale for the drugs used in hair transplantation surgery? AB - BACKGROUND: Various medications may be used before, during, or after hair transplantation surgery (HTS) with the aims of maximizing patient comfort, reducing unwanted side effects, and improving the results of HTS. OBJECTIVE: The objectives of this study were to determine the current practice pattern and rationale for drug prescribing by a group of leading hair transplant surgeons and to review the literature for the evidence upon which these prescribing patterns were based. METHODS: A postal questionnaire was sent to 16 hair transplant surgeons from the United States and Canada, and the answers were analyzed. The relevant evidence-based literature concerning HTS was reviewed by medicine search. RESULTS: Questionnaires suitable for analysis were received from 14 of the surgeons. There were many differences in the pattern of prescribing drugs for the HTS procedure. There was general agreement about the use of local anesthetics but no consensus about the withholding of agents that might increase bleeding; the use of pre- and postoperative analgesics; the use of topical and systemic antibiotics; the use of corticosteroids; or minoxidil. Randomized controlled studies relating to these issues for HTS were not identified in the literature. CONCLUSION: A lack of consensus exists about the drugs used in HTS based on a lack of evidence-based medicine. PMID- 9754085 TI - The new fluor-hydroxy pulse peel. A combination of 5-fluorouracil and glycolic acid. AB - BACKGROUND: Chemical peels have become an increasingly popular method to treat a myriad of benign skin disorders. Individually, glycolic acid (GA) and 5 fluorouracil (5-FU) have been proven efficacious in the treatment of actinically damaged skin. However, to our knowledge the literature lacks a study examining the synergistic effects of these two agents in the treatment of actinic keratoses (AKs) and solar damage. OBJECTIVE: The aim of the study was to determine if a combination of 5-FU and 70% GA, when delivered in pulse doses, would have greater efficacy than using GA alone in destroying precancerous AKs and improving the cosmetic appearance of the skin. METHODS: A prospective randomized controlled study was designed with 18 subjects who had clinically apparent facial AKs. Each patient was treated with the combination of 5-FU and GA to one half of the face, while GA alone was applied to the other half, in a randomized fashion. A before treatment count of the number of AKs present on each half of the face was recorded and pretreatment photographs were taken. The solutions were applied weekly to all patients for an 8-week period. A posttreatment count of AKs on each half of the face along with posttreatment photographs followed at 6 months. RESULTS: The combination of 5-FU and GA cleared 91.94% of AKs at a 6-month follow up period as compared with 19.67% clearing by GA alone. There were no significant side effects reported with the combination peel. CONCLUSION: The fluor-hydroxy pulse peel applied in a pulse dose regimen not only provides cosmetic improvement, but more importantly, has a therapeutic effect on ablating premalignant AKs. This therapeutic effect occurs without the usual morbidity associated with using 5-FU alone in a nonpulsed dosage. Additionally, it is evident that the superficial peeling induced by alpha hydroxy acids may improve cosmesis of actinically damaged skin, but the GA alone cannot destroy a significant number of AKs. PMID- 9754086 TI - Characteristics of office-based visits for skin cancer. Dermatologists have more experience than other physicians in managing malignant and premalignant skin conditions. AB - BACKGROUND: Actinic keratoses and skin cancer constitute a major public health problem for predisposed individuals. OBJECTIVE: The purpose of this paper is to determine the characteristics of office-based visits for actinic keratoses and skin cancer in the United States. METHODS: The National Ambulatory Medical Care Survey provided data on office-based physician visits for actinic keratoses and skin cancer in 1993 and 1994. RESULTS: There were 3.7 million visits per year for actinic keratoses, 3.1 million visits per year for nonmelanoma skin cancer (NMSC), and 430,000 visits per year for melanoma. Excisions and destructions of lesions accounted for 90%, 67%, and 62% of procedures for actinic keratoses, NMSC, and melanoma, respectively. Dermatologists (683), plastic surgeons (37), and general and family physicians (11) managed more visits per physician per year than other specialists. CONCLUSION: Dermatologists have significantly more experience managing skin cancer than do other physicians. PMID- 9754087 TI - A precision machine for mounting tissue for Mohs micrographic surgery. AB - BACKGROUND: Mounting specimens onto chucks, so the cryostat knife follows the exact path of the surgeon's knife, poses difficulties even for experienced histotechnologists. Oftentimes, the frozen planar surface of the tissue falls at an angle that requires gimballing the chuck holder to compensate. OBJECTIVE: To demonstrate the use of a precision tissue mounting machine and associated fast freezing chucks. METHODS: Specimens of pickled pig skin were marked with blue dye on the cut surface. The pieces were mounted onto chucks with the blue surface exposed, and then sectioned in a cryostat. RESULTS: A complete section with all edges was obtained after eight turns of the cryostat handwheel, at 12 microns per turn. CONCLUSIONS: A precision machine used in conjunction with a properly adjusted chuck holder can yield results unobtainable by other methods. PMID- 9754088 TI - A comparison of frozen and paraffin sections in dermatofibrosarcoma protuberans. AB - BACKGROUND: Dermatofibrosarcoma protuberans (DFSP) is a tumor with a high local reoccurrence rate. Mohs micrographic surgery offers the highest cure rate. However, differentiating minimal residual tumor from normal skin can be difficult during Mohs surgery. OBJECTIVE: To clarify the problem of determining when a tumor-free plane had been achieved during Mohs surgery for a DFSP. METHODS: In two patients with DFSPs, we compared frozen and paraffin-embedded sections extending from tumor to normal skin, using both H&E and CD34 stains. RESULTS: On frozen, but not paraffin-embedded, sections scattered dermal spindle cells were seen in normal skin. CONCLUSIONS: Scattered dermal spindle cells in the dermis of normal skin make it difficult to differentiate minimal residual tumor from normal dermis during Mohs surgery. A biopsy of normal skin can be useful as a control in this setting. PMID- 9754089 TI - Mohs micrographic surgery using HMB-45 for a recurrent acral melanoma. AB - BACKGROUND: Acral melanomas are uncommon. Due to the thick overlying stratum corneum, accurate estimation of margins is difficult for minimally pigmented or amelanotic melanomas on the palm or sole. OBJECTIVE: To describe the use of Mohs micrographic surgery using frozen sections and HMB-45 immunostaining in the treatment of a multiply recurrent acral melanoma that had failed both standard surgery and Mohs surgery. METHODS: The melanoma was excised by Mohs technique, and the margins were checked by frozen section and HMB-45 immunostaining. RESULTS: The melanoma was completely excised in 11 stages, resulting in a defect that covered much of the plantar surfaces of the ball of the left foot, great, second, third, fourth, and fifth toes. No recurrence has been noted in 22 months of follow-up. CONCLUSIONS: HMB-45 immunostaining is a very valuable adjunct to examination of surgical margins for melanoma, particularly when combined with such histologic features as clustering of cells, melanocyte position within the epidermis, and cytologic atypia. PMID- 9754090 TI - Nasal valve dysfunction after Mohs surgery for skin cancer of the nose. AB - BACKGROUND: The nasal valve is the narrowest part of the nasal vestibule and is an important regulator of airflow. Nasal valve insufficiency (NVI), which can be caused by nasal surgery, results in nasal stuffiness, or resistance to airflow on inspiration. This entity has not been well described in the dermatologic surgery literature. OBJECTIVE: To study nasal valve insufficiency in 100 consecutive patients who had Mohs surgery for skin cancer of the nose, review the literature, and report methods for prevention of this complication. METHODS: One hundred consecutive nasal Mohs surgery cases were studied retrospectively. Symptomatic patients were evaluated based on several parameters to determine causative and exacerbating factors, and possible methods of prevention. The pertinent literature was reviewed. RESULTS: Out of 100 patients, 92 responded--38 (41.3%) of whom were determined to be "at risk" for NVI based on the anatomic location of their defects. Out of those 38, five (13.2%) had new onset nasal stuffiness. An additional three of the 38 (7.9%) reported an exacerbation of prior nasal obstructive symptoms. Healing by secondary intention, bulky flaps, inadequate cartilaginous support, inappropriate choice of flap, mucosal scarring, and sacrifice of nasalis and levator labii superioris alaeque nasi fibers were identified as contributors to postoperative NVI. CONCLUSION: NVI is a relatively common complication of Mohs surgery and reconstruction of the lower third of the nose. Treatment is difficult, but prevention is possible in many instances. Therefore, surgeons should be well aware of this entity and techniques that may aid in its prevention. PMID- 9754091 TI - Experimental rationale for treatment of high-risk human melanoma with zinc chloride fixative paste. Increased resistance to tumor challenge in murine melanoma model. AB - BACKGROUND: Fixed-tissue micrographic surgery (Mohs) of melanoma has been shown by retrospective analysis to improve 5-year survival. OBJECTIVES: To determine whether zinc chloride fixative paste acts as an immune adjuvant to increase host resistance to melanoma. METHODS: We performed a murine study using the poorly immunogenic B16 melanoma of C57Bl6J mice, and the more immunogenic K1735p melanoma of C3H/HeN mice. Tumors were treated with zinc chloride paste and excised 24 hours later (Group 1), or simply excised (Group 2). Mice were challenged 7 days later with injection of melanoma cells at a distant site, and tumor growth in this second site was followed. RESULTS: K1735p melanomas developed at the challenge site in 69% of mice treated with excision versus 32% of mice treated with zinc chloride fixation (P < 0.025). Development of B16 melanoma was not altered by zinc chloride fixation. CONCLUSION: Zinc chloride fixation of the more immunogenic K1735p melanoma increased resistance to subsequent tumor challenge, suggesting that zinc chloride fixative paste acts as an immune adjuvant. PMID- 9754092 TI - Treatment of xanthelasma palpebrarum with bichloracetic acid. AB - BACKGROUND: Although many treatment modalities have been described for xanthelasma palpebrarum, no single technique has emerged as dominant. OBJECTIVES: Our purpose was to review the various therapeutic modalities for xanthelasma and to assess the efficacy of topical bichloracetic acid. METHODS: Thirteen patients with 25 xanthelasma were treated with topical 100% bichloracetic acid. Efficacy was assessed over a follow-up period of 7 months to 10.5 years (average, 64 months). RESULTS: Eighty-five percent of patients experienced initial complete clearing, and 72% of their lesions have not required retreatment over an average period of 68 months. Recurrences responded well to repeat treatment. Eighty-three percent of recurrent or poorly responsive lesions were associated with high cholesterol. The most resistant patient had four-lid involvement. Excellent cosmetic results and high patient satisfaction were seen. CONCLUSIONS: Topical bichloracetic acid is a viable alternative to other modalities in the management of xanthelasma. Advantages include simplicity, cost-effectiveness, speed, safety, and efficacy. PMID- 9754093 TI - Treatment of a Becker's nevus using a 694-nm long-pulsed ruby laser. AB - Becker's nevus is an uncommon pigmented smooth muscle hamartoma that develops during adolescence and occurs primarily in young men. The nevus is characterized by hypertrichosis and hyperpigmentation and is usually located unilaterally over the shoulder, upper arm, and scapula. We describe a patient with a Becker's nevus who was treated with a long-pulsed ruby laser in order to decrease hair density and pigmentation. PMID- 9754094 TI - Statement on ultrasonic liposuction. Task Force on Ultrasonic Liposuction of the American Society for Dermatologic Surgery. PMID- 9754095 TI - Sclerosing lipogranuloma secondary to supposed vitamin E injection for facial rejuvenation: successful treatment with intralesional steroids. PMID- 9754096 TI - The effects of apnea on timing examinations for optimization of gadolinium enhanced MRA of the thoracic aorta and arch vessels. AB - PURPOSE: Our purpose was to determine the effects of apnea during end-inspiration compared with free breathing on timing examinations performed to optimize gadolinium-enhanced 3D MR angiography (MRA) of the thoracic aorta and arch vessels. METHOD: Thirty patients referred for gadolinium-enhanced 3D MRA of the thoracic aorta and branch vessels underwent two timing examinations: one performed during free breathing and one during apnea at end-inspiration to replicate more closely the respiratory pattern used to obtain 3D MRA. For each, axial images at the level of the proximal neck were acquired every 2 s for 40 s, during which time 1 ml of gadolinium contrast agent followed by 20 ml of saline was infused at 2 ml/s. The time to peak arterial enhancement (Ta), time to first jugular venous enhancement (Tj), and arterio-venous window (time from peak arterial enhancement to first jugular venous enhancement; AV) were compared for the two examinations in each patient. RESULTS: Overall there was no statistically significant difference in Ta, Tj, or AV between examinations performed during free breathing and apnea in end-inspiration, although a trend to delayed circulation times was observed with apnea (p = 0.2-0.3). In five patients (17%), the difference in Ta between free breathing and apnea was 4 s; in three patients (10%), the difference was 6 s. CONCLUSION: Circulation times determined during apnea at end-inspiration may differ from those obtained during free breathing by as much as 6 s in an individual patient. This difference may account for inappropriately timed gadolinium-enhanced MR angiograms performed with timing examinations, especially when very short acquisition times and low doses of gadolinium (20 ml) are used. PMID- 9754097 TI - Coronary artery anomalies with a shunt: evaluation with electron-beam CT. AB - PURPOSE: Our goal was to evaluate the role of electron-beam CT (EBT) in the diagnosis of patients with coronary artery anomalies with a shunt. METHOD: We performed EBT in seven patients with coronary artery anomalies with a shunt. Four cases were coronary artery fistula (CAF) and three were an anomalous origin of the left coronary artery from the pulmonary artery (ALCA from PA). Serial single volume mode scanning was performed at end-diastole to evaluate the anatomical course of the anomalous coronary arteries. Cine mode scanning was done in all but one to examine the ventricular wall motion and volumetrics. RESULTS: EBT could detect the course and drainage sites of all CAFs and ALCAs from PA. Cine mode scanning revealed reduced wall motion in one case with CAF and two cases with ALCA from PA. CONCLUSION: EBT serves a useful role in the assessment of coronary artery anomalies with a shunt. PMID- 9754098 TI - Primary angiosarcoma of the pulmonary arteries: dynamic contrast-enhanced MRI. AB - PURPOSE: Our goal was to assess the MR appearance and histologic correlation of primary pulmonary artery angiosarcoma. METHOD: Four patients with tumorous masses in the pulmonary arteries were evaluated by dynamic contrast-enhanced MRI using T1- and T2-weighted SE images, GE images, as well as coronal 3D MRA in breath hold technique. The percentage of tumor enhancement was determined by measuring regions of interest before and after Gd-DTPA administration on the 2D multiplanar spoiled GRE images. RESULTS: All four masses showed some contrast enhancement on the dynamically acquired GRE images. The degree of contrast enhancement correlated with the degree of tumor differentiation, content of myxoid matrix, and associated thrombus. Contrast-enhanced 3D MRA was useful for preoperative delineation of the peripheral pulmonary arteries to the subsegmental order. CONCLUSION: Dynamic contrast-enhanced 3D MRA of the pulmonary arteries can be used to delineate pulmonary arterial angiosarcomas preoperatively. Considerable variability of contrast agent uptake reflects the wide histologic behavior of these masses in differentiation from central pulmonary embolism. PMID- 9754099 TI - CT diagnosis of an unusual aortic dissection with intimointimal intussusception: the wind sock sign. AB - A unique combination of CT findings is reported in a rare case of aortic dissection with intimointimal intussusception. The CT showed a wind sock-like appearance in the contrast column of the aortic arch, which was felt to be characteristic of the intussuceptum. Complementary CT findings, including proximal flap in the dilated root of the aorta, no mid-ascending aortic flap, a descending aortic flap, and pericardial effusion, enabled establishment of the preoperative diagnosis. PMID- 9754100 TI - The spotted spleen: CT and clinical correlation in a tertiary care center. AB - PURPOSE: The goal of our study was to examine the prevalence of multiple hypodense splenic nodules and their associated diagnoses and to correlate CT appearance with clinical presentation and diagnosis. METHOD: Records of all patients undergoing contrast-enhanced CT from July 1994 through September 1997 were reviewed. Charts and CT scans of patients with multiple (more than five) hypodense splenic nodules were then evaluated. RESULTS: During the search period, there were 8,764 patients examined. Multiple hypodense splenic nodules were identified in 45 patients. Sixteen patients had malignant neoplasia as an etiology, with two patients having a benign tumor. Ten patients had an infectious etiology; nine patients had an inflammatory but noninfectious etiology; in eight patients, a diagnosis was not established; five of these patients were followed for > 18 months. CONCLUSION: Multiple hypodense splenic nodules are uncommon. Lymphoma, infection, and sarcoid were the three most common disorders in the symptomatic patient, with infection strongly correlated with a compromised immune system. In the asymptomatic patient, nonlymphomatous metastatic disease, benign tumor, and sarcoid were most common. Although overlap exists between diagnostic groups, lymphoma tends to have larger, more variable nodules, whereas infection tends to occur with smaller, more uniform nodules. Sarcoid is intermediate in appearance. PMID- 9754101 TI - Differentiation of hypervascular hepatic pseudolesions from hepatocellular carcinoma: value of single-level dynamic CT during hepatic arteriography. AB - PURPOSE: The purpose of our study was to assess the efficacy of single-level dynamic CT during hepatic arteriography (D-CTA) in the differentiation between hypervascular hepatocellular carcinoma (HCC) and hypervascular pseudolesion. METHOD: D-CTA was performed in nine cases with HCC and nine cases with pseudolesion. Findings on D-CTA were retrospectively analyzed. RESULTS: The transition of the stain of pseudolesion on D-CTA was divided into three phases: (1) inflow of the contrast material into the portal vein within the lesion, (2) lesion staining, and (3) fading out of the stain; that of HCC was divided into four phases: (1) inflow of CM into tumor, (2) tumor staining, (3) inflow of CM into the adjacent liver, and (4) coronal stain of adjacent liver. The coronal stain was seen in all HCCs but not in any pseudolesions. CONCLUSION: The present study suggest that D-CTA is a helpful option in the differentiation between HCC and pseudolesion. PMID- 9754102 TI - The role of 3D spiral CT in early gastric carcinoma. AB - PURPOSE: The purpose of this study is to assess the role of 3D imaging using spiral CT for the detection and evaluation of early gastric carcinoma (EGC). METHOD: Thirty-one patients with EGC underwent 3D and axial imaging using spiral CT, and all cases were confirmed at surgery. According to the pathologic specimens, one patient (3.2%) had type I, five (16.1%) had type IIb, 13 (42.0%) had type IIc, three (9.6%) had types IIc + IIb, three (9.6%) had types IIc + III, two (6.5%) had types IIa + IIc, two (6.5%) had types IIb + IIc, and two (6.5%) had types IIb + III. Spiral CT was first performed with 3 mm collimation, 4.5 mm/s table feed, and a 1 mm reconstruction interval in the supine position after intravenous injection of Buscopan and ingestion of gas. The 3D shaded surface display renderings were performed, analyzed, and graded as excellent, good, or poor. Axial CT scanning was performed with 5 mm collimation, 7 mm/s table feed, and a 5 mm reconstruction interval in the prone position after ingestion of water. RESULTS: With use of axial CT images, 20 of 31 tumors (64.5%) were detected. Tumors were suspected in two cases, but nine were not detected. With use of 3D images, 29 of 31 tumors (93.5%) were detected (p < 0.05). Of the 31 cases of EGC, excellent 3D images were obtained in 6 patients (19.3%), good 3D images in 21 (67.7%), and poor 3D images in 2 (6.5%). In the two cases with poor images, the tumor was confined to the mucosal layer and the types were EGC IIc + IIb. In the remaining two cases (6.5%), the tumors were not detected by 3D images. In one case, the tumor was confined to the mucosal layer, and in the other it was located in the pyloric antrum. Both tumors were EGC type IIc. The two cases not detected by 3D imaging were also not detected by axial CT scanning. CONCLUSION: The detection rate of EGC is higher using 3D imaging than axial CT scan alone. PMID- 9754103 TI - Dynamic gadolinium-enhanced MR findings in infantile hepatic hemangioendothelioma. AB - We report a case of a 2 1/2-year-old girl presenting with abdominal pain, fever, vomiting, and hepatomegaly. In spite of the unusual age at presentation, dynamic gadolinium-enhanced MR findings, which have not been previously illustrated, proved to be highly specific for the diagnosis of infantile hepatic hemangioendothelioma because of the characteristic enhancement pattern. PMID- 9754104 TI - MRI of hydatid disease of the liver: a variety of sequences. AB - Hydatid disease is a parasitic manifestation that is most commonly seen in the liver. Diagnosis of this condition in the liver is usually straightforward and achieved through US and CT. Complicated cases and the often bizarre appearance of the disease can frequently create problems for the diagnostician, however. MRI has become an important diagnostic tool in the evaluation of complicated liver masses, with routine application of fat-suppressed, fast, and breath-hold techniques. In this pictorial essay, we demonstrate various features of hydatid disease using these new methods. PMID- 9754105 TI - Spontaneous hemorrhage of abdominal splenosis. PMID- 9754106 TI - A case of congenital absence of the intrahepatic portal vein diagnosed by MR angiography. PMID- 9754107 TI - Spontaneous intracholecystic hemorrhage due to polyarteritis nodosa. PMID- 9754108 TI - Interobserver variability in the interpretation of unenhanced helical CT for the diagnosis of ureteral stone disease. AB - PURPOSE: The purpose of this study was to analyze interobserver agreement in the interpretation of unenhanced helical CT (UHCT) for the evaluation of ureteral stone disease and obstruction. METHOD: One hundred three UHCT examinations were independently and retrospectively reviewed by five readers including attending radiologists, a radiology resident, and an attending urologist. Examinations were interpreted as positive, negative, or indeterminate for ureteral stone disease and obstruction. The Cohen kappa test was used to measure interobserver agreement. The accuracy of the readers was also assessed. RESULTS: The kappa value ranged from 0.67 to 0.71 among the three attending radiologists and from 0.65 to 0.67 among the radiology attending physicians and radiology resident. Although the urologist tended to agree less well with the other readers (kappa range: 0.33-0.46), there was no statistically significant difference (p < 0.05) in the accuracy among all five readers. The percentage of cases interpreted as indeterminate ranged from 8 to 25% and almost invariably involved difficulty distinguishing phleboliths from minimally obstructing distal ureteral calculi. The percentage of UHCT scans correctly interpreted as positive and correctly interpreted as negative ranged from 73% (n = 27) to 86% (n = 32) and 63% (n = 22) to 86% (n = 30), respectively. CONCLUSION: Interobserver agreement was very good among the radiology attending physicians and resident and moderate with the urologist. The examination is an accurate technique in the evaluation of ureteral stone disease, although limitations exist, particularly in the diagnosis of minimally obstructing distal ureteral calculi. PMID- 9754109 TI - MR appearance of uterine dehiscence in the post-cesarean section patient. AB - PURPOSE: CT has been shown to be unreliable for detecting uterine dehiscence in the postoperative period after cesarean section (c-section). The purpose of this investigation is to describe the MR appearance of uterine dehiscence in this setting and identify features that distinguish complete from partial dehiscence. METHOD: Over an 82 month period, all charts and MR reports of patients that underwent MRI of the pelvis after c-section were reviewed for uterine dehiscence. Altogether, 55 patients were imaged. Positive cases for dehiscence were retrospectively reviewed by two radiologists. Imaging criteria for complete dehiscence consisted of transmural disruption. Criteria for partial dehiscence consisted of disruption of the endometrial and/or serosal layer, without transmural extension. RESULTS: On MRI, five patients demonstrated abnormalities suggestive of incisional dehiscence. Based on these imaging criteria, two of these showed complete dehiscence that was proven at surgery and three showed findings of partial dehiscence. The optimal imaging plane was perpendicular to the incision. CONCLUSION: MR features may be utilized to identify total uterine dehiscence and may be more effective than CT. PMID- 9754110 TI - High-resolution contrast-enhanced MRI of the uterus with a phased-array multicoil. AB - High-resolution contrast-enhanced dynamic MRI of the uterus can be performed with the combination of a phased-array multicoil and fast GE techniques. This technique can improve the ability to visualize normal anatomy of the uterus and periuterine tissues, including vascular structures and pelvic ligaments, and to detect pathologic processes of the uterus and determine their extent. PMID- 9754111 TI - MR defecography: depiction of anorectal anatomy and pathology. AB - With the advent of open-configuration MR systems, enabling image acquisition in a vertical patient position, MR defecography has become possible. MR defecography permits analyses of the anorectal angle, opening of the anal canal, functioning of the puborectal muscle, and descent of the pelvic floor during defecation. The rectal walls are well delineated on the GRE images, permitting visualization of intussusceptions and rectoceles. The concomitant depiction of structures surrounding the anorectal canal is helpful in the assessment of a spastic pelvic floor and the descending perineum syndrome and in permitting visualization of enteroceles. Dynamic MR defecography is an attractive alternative in the evaluation of defecation disorders. PMID- 9754112 TI - Sister Mary Joseph nodule from metastatic renal cell carcinoma. AB - Metastatic lesions of the umbilicus are more common than primary malignancies and are commonly referred to as Sister Mary Joseph nodules. Most arise from the stomach or the female genital tract. We describe an unusual case of renal cell carcinoma with peritoneal implants producing a Sister Mary Joseph nodule diagnosed by CT. To our knowledge, no report of a urinary tract malignancy with umbilical involvement has been described in the radiological literature. PMID- 9754113 TI - Tumor in ectopic omental ovary in Mayer-Rokitansky-Kuster-Hauser syndrome: CT findings. PMID- 9754114 TI - Increased brain activation in response to odors in patients with hyposmia after theophylline treatment demonstrated by fMRI. AB - PURPOSE: Our goal was to demonstrate that medical therapy in patients with smell loss (hyposmia) that restored olfactory function toward or to normal could be verified and quantitated by functional MRI (fMRI) of brain and that visual representation of these changes could be used to identify these patients. METHOD: Multislice FLASH MR or echo planar MR brain scans were obtained in four patients with hyposmia in response to three olfactory stimuli both before and after treatment with theophylline. Activation images were derived using correlation analysis, and ratios of brain area activated to total brain area were obtained. RESULTS: Prior to treatment, all patients stated that they could not smell; these losses were confirmed by standard psychophysical tests. At this time, fMRI brain activation in response to odors was significantly less than that measured in normal volunteers and similar to activation measured previously in other patients with a similar type of hyposmia. After treatment, subjective smell function improved in three patients and no improvement occurred in one; results were confirmed by psychophysical tests. In each patient in whom smell acuity improved, brain activation in response to each odor increased in each section and mean activation increased significantly for each odor. Activation increased in all regions previously associated with olfactory stimulation and was particularly apparent in orbitofrontal cortex, frontal lobe component of cingulate gyri, temporal lobe gyri, and hippocampus. There also was consistent activation in superior, middle, and inferior frontal lobe gyri. There were no changes in brain activation after treatment in the patient in whom smell did not improve. CONCLUSION: These results demonstrate that theophylline is an effective therapeutic agent to correct hyposmia in some patients with smell loss. These changes have been documented by fMRI brain scans using olfactory stimuli. This is the first study in which this type of objective improvement following medical treatment has been demonstrated in patients with hyposmia. PMID- 9754115 TI - Gadolinium-enhanced fat-suppressed T1-weighted imaging of the head and neck: comparison of gradient and conventional SE sequences. AB - PURPOSE: The purpose of this study was to compare contrast-enhanced GRE and conventional SE (CSE) fat-suppressed T1-weighted techniques in the evaluation of head and neck lesions. A hybrid, opposed phase, frequency-selective, fat suppressed fast multiplanar spoiled GRE (FMPSPGR) sequence was compared with a fat-suppressed CSE sequence. METHOD: Thirty-two patients with head and neck pathology were evaluated with both fat-suppressed CSE and FMPSPGR sequences. Regions of interest obtained by two viewers in consensus were used to establish contrast-to-noise (CNR) and signal-to-noise ratios for both sequences. Three neuroradiologists also independently reviewed the images for quality of fat suppression, lesion conspicuity, and potential pitfalls. RESULTS: The CNR of the FMPSPGR sequence was superior to that of the fat-suppressed CSE sequence. Subjectively, all three reviewers rated the FMPSPGR sequence as having fat suppression equal to or better than that in the CSE sequence in 94% of cases. Imaging times for the FMPSPGR sequence were 60-75% faster than those for the CSE sequence. CONCLUSION: Enhanced imaging of the head and neck region using an opposed phase, fat-suppressed GRE sequence results in improved fat suppression compared with the CSE technique, with substantial savings in imaging time. PMID- 9754116 TI - Image generation of serotonin synthesis rates using alpha-methyltryptophan and PET. AB - PURPOSE: We sought to create functional images of the serotonin (5-HT) synthesis rate obtained with alpha-[11C]methyl-L-tryptophan (alpha-MTrp) and PET and standardize them into the stereotaxic coordinate system. METHOD: Dynamic PET scans were performed in 11 healthy subjects after an injection of alpha-MTrp. Results obtained by the Patlak plot and nonlinear least-squares methods using arterial plasma as input function were compared. The pixel-by-pixel calculation of functional images of 5-HT synthesis was done by the Patlak plot approach, after results were compared by two methods. Input function obtained by combining venous plasma and sinus radioactivities was also evaluated as an alternative to arterial input function. RESULTS: There were no significant differences in 5-HT synthesis between the two calculation methods. The normalized venous input functions gave similar results as the arterial input function. The regional rates of 5-HT synthesis from functional images were not significantly different from those obtained by graphical plotting. CONCLUSION: alpha-MTrp images could be converted into functional images representing 5-HT synthesis rates in the living brain, facilitating statistical comparison. PMID- 9754117 TI - The effect of xenon inhalation speed on cerebral blood flow obtained using the end-tidal method in xenon-enhanced CT. AB - PURPOSE: The purpose of this work is to show how variations in inhalation speed of xenon gas affect cerebral blood flow (CBF) values obtained using the end-tidal method on xenon-enhanced CT (Xe-CT). We tried to clarify whether arterial xenon concentration could keep up with end-tidal xenon concentration by evaluating the effect of xenon inhalation speed on calculated CBF values. METHOD: The same subject underwent two or three consecutive Xe-CT examinations, varying xenon inhalation speed. The rate constants of applied inhalation speeds were 0.1-0.15 min-1 (low speed), 0.25-0.3 min-1 (middle speed), and 1-2 min-1 (high speed), respectively. RESULTS: No significant difference was observed among the CBF values of the same subject obtained under different inhalation speeds. CONCLUSION: End-tidal xenon can closely reflect arterial xenon under the customary method of xenon supply. The end-tidal method can provide reliable absolute CBF values, assuming actual CBF values are substantially unchanged regardless of the inhalation speed variation applied in this work. PMID- 9754118 TI - Calculation of apparent diffusion coefficients (ADCs) in brain using two-point and six-point methods. AB - We evaluated the relative accuracy of calculating apparent diffusion coefficients (ADCs) of intracranial tissues using a two-point versus a six-point regression technique. Echo planar diffusion-weighted MRI was performed at 1.5 T in three standard locations and in pathologic regions in 10 subjects using gradient strengths corresponding to b values of 1, 100, 200, 500, 800, and 1,000 s/mm2. Estimation of ADCs was made using two methods: a nonlinear regression model using measurements from the full set of b values (six-point technique) and linear estimation using b values of 1 and 1,000 only (two-point technique). A high correlation between the two methods was noted (R2 = 0.999), and the mean percentage difference was 0.84%. These results suggest there is little error in estimating brain ADCs using a two-point technique. PMID- 9754119 TI - Herpes simplex virus 1 pneumonia: patterns on CT scans and conventional chest radiographs. AB - PURPOSE: The goal of our study was to describe the herpes simplex virus type 1 (HSV 1) pneumonia patterns on CT scans and chest radiographs. METHOD: We retrospectively reviewed clinical records and chest radiographs of 24 patients with HSV 1 pneumonia and 10 with pneumonia from combined HSV and mixed flora infection. We also reviewed CT scans available for eight patients with HSV pneumonia and four with mixed pneumonia. RESULTS: CT scans of eight patients with HSV pneumonia demonstrated multifocal segmental and subsegmental ground-glass opacities (n = 8), additional focal areas of consolidation (n = 6), scattered distribution (n = 6), and pleural effusions (n = 7). Chest radiographs (23 patients) showed patchy segmental and subsegmental ground-glass opacities and consolidation (n = 23), scattered distribution (n = 20), and pleural effusions (n = 12). Radiographic patterns for isolated HSV pneumonia and mixed flora pneumonia were not significantly different. CONCLUSION: With a growing population of at risk immunosuppressed patients, it is important to recognize CT and chest radiography patterns consistent with, although nonspecific for, HSV 1 pneumonia. PMID- 9754120 TI - Diffuse pulmonary lesions in early phase Takayasu arteritis predominantly involving pulmonary artery. AB - We report a case of early phase Takayasu arteritis that predominantly involved the pulmonary arteries and had, along with expected radiographic findings, the unusual pattern of diffuse pulmonary parenchymal lesions on CT. We suggest that this finding may be an additional feature in early phase Takayasu arteritis and need not require investigation for an additional diagnosis. PMID- 9754121 TI - Cushing syndrome due to a pulmonary carcinoid tumor: multimodality imaging and diagnosis. PMID- 9754122 TI - Ligaments of the ankle: normal anatomy with MR arthrography. AB - PURPOSE: Our purpose was to define the normal MR arthrographic anatomy of ankle ligaments. METHOD: Prior to injection of intraarticular gadolinium in cadaveric ankle joints, proton density and T2-weighted images were obtained to assess the integrity of the ligaments and tendons as well as the amount of preexisting joint effusion. Following injection of 10 ml of contrast agent (gadopentetate dimeglumine 1:250, Omnipaqe 300, Knox gelatin 50%, and methylene blue), T1 weighted images with fat saturation in axial, oblique axial, coronal, and sagittal planes were obtained in neutral, dorsiflexion, and plantar flexion positions. Specimens were sectioned, allowing anatomic and MR correlation. RESULTS: Contrast agent outlining anterior and posterior aspects of the anterior talofibular ligament and posterior talofibular ligament (PTAF) was a normal finding, related to anterior and posterior recesses of the ankle joint that extend out beyond these ligaments in an anteroposterior direction above the level of the ligaments. Intraarticular contrast material allowed resolution of superficial and deep components of the posterior tibiofibular ligament. Both were seen separately from PTAF with dorsiflexion. Posterior intermalleolar ligament was not present in our specimens. Visualization of calcaneofibular ligament was much improved by contrast material outlining the articular aspect of the ligament. Visualization of the syndesmotic ligamentous complex also was improved by contrast material outlining the articular side of the ligaments and separating them from adjacent bone. Superiorly, the distribution of contrast agent was limited by the interosseous ligament. Visualization of the medial collateral ligaments was not improved by the presence of the intraarticular contrast material. CONCLUSION: MR arthrography of the ankle allows improved visualization and evaluation of the lateral and syndesmotic ligamentous complex. PMID- 9754123 TI - The plica syndrome: diagnostic value of MRI with arthroscopic correlation. AB - PURPOSE: Our goal was to evaluate the diagnostic value of MRI in plica syndrome. METHOD: MR images of a patient group (n = 55) with arthroscopically confirmed pathologic mediopatellar plicae were retrospectively analyzed and compared with those of a control group (n = 100). We obtained axial multiplanar gradient recalled (MPGR), axial T1-weighted, and sagittal T2-weighted MR images. MR images were assessed for the width and length of all medial plicae. RESULTS: In the diagnosis of plica syndrome, sensitivity and specificity were 73 and 78% on axial MPGR images, 71 and 83% on sagittal T2-weighted images, and 95 and 72% on combination of both images, respectively. The incidence of pathologic medial plica increased with a criterion of extension beyond the medial end of the patella on axial MPGR images. CONCLUSION: MRI is a useful screening method in the diagnosis of plica syndrome. PMID- 9754124 TI - Malignant fibrous histiocytoma of soft tissue imaging with emphasis on MRI. AB - Malignant fibrous histiocytoma (MFH) is the most common soft tissue sarcoma in adults. This pictorial essay describes and illustrates the clinical, pathologic, and radiologic features of MFH. The cross-sectional imaging features on CT and MRI are emphasized in relation to the diagnosis and staging of MFH. PMID- 9754125 TI - An image-processing system for qualitative and quantitative volumetric analysis of brain images. AB - In this work, we developed, implemented, and validated an image-processing system for qualitative and quantitative volumetric analysis of brain images. This system allows the visualization and quantitation of global and regional brain volumes. Global volumes were obtained via an automated adaptive Bayesian segmentation technique that labels the brain into white matter, gray matter, and cerebrospinal fluid. Absolute volumetric errors for these compartments ranged between 1 and 3% as indicated by phantom studies. Quantitation of regional brain volumes was performed through normalization and tessellation of segmented brain images into the Talairach space with a 3D elastic warping model. Retest reliability of regional volumes measured in Talairach space indicated errors of < 1.5% for the frontal, parietal, temporal, and occipital brain regions. Additional regional analysis was performed with an automated hybrid method combining a region-of interest approach and voxel-based analysis, named Regional Analysis of Volumes Examined in Normalized Space (RAVENS). RAVENS analysis for several subcortical structures showed good agreement with operator-defined volumes. This system has sufficient accuracy for longitudinal imaging data and is currently being used in the analysis of neuroimaging data of the Baltimore Longitudinal Study of Aging. PMID- 9754127 TI - Aunt Minnie's corner. Left paraduodenal internal hernia. PMID- 9754126 TI - Modeling brain deformations in Alzheimer disease by fluid registration of serial 3D MR images. AB - Serial 3D MR scan pairs (1 year intervals) from nine control subjects and nine Alzheimer disease (AD) patients were matched using an accurate fast fluid algorithm. Subtraction of matched scans indicated near complete cancellation of brain structure; mean cross-correlation coefficients were 0.946 (controls) and 0.933 (AD). In the AD group, regions of expansion and contraction in the deformation field were consistent with the known patterns of tissue loss and expansion of CSF spaces. Fluid registration may permit better localization of structural change. PMID- 9754128 TI - Aging and the liver: an update. AB - The issue of whether or not liver function is compromised in the elderly population remains unresolved. Numerous age-related changes in hepatic structure and function have been described, but many of these observations are qualitative, were made under suboptimal experimental conditions, or are simply contradictory. Changes in hepato-cellular structural parameters, e.g., increased hepatocyte size, increase in the number of binucleated cells, altered mitochondria, and endoplasmic reticulum, have been reported. However, quantitative morphological analyses have refuted many of these observations. There are few functional data that correlate with structural changes. Serum and biliary cholesterol appear to rise, predisposing elderly people to increased incidences of coronary disease and gallstones, respectively. The rate of liver regeneration declines in old animals, but the regenerative capacity remains unchanged, perhaps reflecting an age associated reduction in the response to hepatotrophic factors. This senescent change has important clinical implications with regard to surgical intervention for liver disease, e.g., resection or transplantation. Nevertheless, most outcomes studies suggest that age alone should not be a determining factor in such clinical decisions. Geriatric patients exhibit a decline in the hepatic clearance of certain drugs and a marked increase in the frequency of adverse drug reactions, reflecting an increase in polypharmacy regimens and declines in liver volume and blood flow rather than reduced Phase I metabolism. Although the livers of elderly subjects are characterized by a decline in adaptive responsiveness and reduced reserve capacity, clinical tests suggest that liver function is well maintained in this age group. PMID- 9754129 TI - Food restriction differentially affects mRNAs encoding the major anterior pituitary tropic hormones. AB - Chronic food restriction (FR) leads to adaptive cellular changes, some of which retard aging. Moreover, some of these changes occur within weeks after onset of FR. Because neuroendocrine mechanisms may mediate these effects, we measured the effect of FR on the messenger ribonucleicacids (mRNAs) encoding all of the tropic hormones of the anterior pituitary (AP). Slot blot and solution hybridization were conducted on AP ribonucleicacid (RNA) samples obtained at 0500 h (AM) and 1500 h (PM) from 3-month-old male Fischer 344 rats fed ad libitum (AL) or FR (60% of AL calories) since 6 weeks of age. PolyA RNA/microgram total RNA was similar in AL and FR rats, indicating that there was no overall effect of FR on mRNA levels. The level of proopiomelanocortin (POMC) mRNA was not reduced by FR when expressed per microgram of RNA or as total AP content. By contrast, the total AP content of the mRNAs encoding LH beta, FSH beta, TSH beta, GH, and PRL was markedly reduced by FR. When expressed per microgram of RNA, however, only GH (AM and PM), FSH beta (AM), TSH beta (PM), and PRL (PM) were reduced by FR. These results reveal that FR differentially affects pituitary tropic hormone mRNA levels within weeks after onset of FR, and are consistent with a role for neuroendocrine alterations in the initiation of adaptive cellular responses to FR. PMID- 9754130 TI - Delayed immune aging in diet-restricted B6CBAT6 F1 mice is associated with preservation of naive T cells. AB - Age-related changes in peripheral blood, spleen, and thymus of ad libitum (AL) fed and dietary restricted (DR) C57BL/6J x CBA/CaH-T6/J F1 (B6CBAT6 F1) mice at young (3 mo), middle (16 mo), and old (30 mo) ages were studied to define how dietary restriction retards immune aging. Dietary restriction at 25% AL intake level initiated at weaning significantly reduced the rates of age-related declines in peripheral blood T helper cells, naive T helper cells, and naive cytotoxic T lymphocytes (CTLs). As a result, concentrations of these cell types in old DR mice were equivalent to 161%, 176%, and 250% of those in old AL controls. Dietary restriction also abolished age-related splenomegaly and decreased total splenocyte numbers in old DR mice. Dietary restriction did not prevent age-related decline in thymus size, but preserved thymus cellularity in old mice. Old DR mice had twice as many total thymocytes and 2.6 times as many CD4+CD8+ immature thymocytes as old AL controls. The correlations between total immature thymocytes and concentrations of circulating naive T helper cells and naive CTLs increase with age and become significant in old mice. Thus, dietary restriction preserves immature T-cell precursors in the thymus during aging to maintain higher concentrations of circulating T helper and naive T cells in peripheral blood. PMID- 9754131 TI - Role of glutathione in the beneficial effect of dietary restriction on bile formation in young, mature, and old rats. AB - We investigated the contribution of bile salts and glutathione (GSH) to the generation of bile flow in young, mature, and old female Sprague-Dawley rats, either fed ad libitum (AL) or subjected to a 40% dietary restriction (DR), which was supplemented or not with vitamins and minerals, starting from weaning. An age related decline in bile flow was observed in the AL group. DR increased bile flow compared to age-matched AL rats, resulting in a twofold increase in the old animals. This was associated with a statistically significantly higher biliary GSH secretion rate and a moderate increase in the bile salt secretory rate. The apparent GSH-dependent flow was significantly increased in DR groups of all ages. Hepatic GSH concentration was closely related to the GSH secretion rate. These results indicate that the increase in biliary GSH content produced by DR is the major mediator of the increased bile flow, resulting in enhanced GSH and GSH derived thiols supply to the intestinal lumen. PMID- 9754132 TI - Stress exacerbates age-related decrements in the immune response to an experimental influenza viral infection. AB - To test the hypothesis that stress exacerbates immune decrements associated with aging, the impact of restraint stress on immunosenescence was assessed using an experimental model of influenza A/Puerto Rico/8/34 viral infection. Beginning one day prior to infection, male C57BL/6 mice, 3 and 22 months of age, were subjected nightly to 12 hours of restraint stress. In both age groups, restraint induced a comparable increase in serum corticosterone levels. However, in contrast to the 3 month-old controls, serum corticosterone levels in 22-month-old mice returned to baseline slower after removal of the stressor. The characteristic influenza driven increase in cellularity of the lung and draining lymph node was decreased by age and further suppressed by stress. Natural killer cell activity and virus specific T helper cell function were also blunted by age and almost completely abrogated by stress. Furthermore, due to the weak immune response to viral infection, aged animals subjected to stress had a lower survival rate than age matched controls. PMID- 9754133 TI - Racial differences in muscle strength in disabled older women. AB - This study examines racial differences in muscle strength, and associations of muscle strength to level of physical activity and severity of disability, among a community sample of 254 black and 665 white, moderately to severely disabled women aged 65 and older. Potential confounders that were adjusted for in the models included age, body weight and height, joint pain, number of chronic conditions, and socioeconomic status. Hand grip, hip flexion, and knee extension forces were measured using portable hand-held dynamometers in the participants' homes. Hand grip strength was measured as the maximal isometric force. Hip flexion and knee extension forces were measured as the greatest force the tester had to apply to break the isometric contraction. A declining strength gradient was observed with increasing severity of disability and for decreasing level of physical activity in both races. At equal levels of disability or physical activity, blacks had better hand grip and hip flexion strength, but knee extension strength did not differ by race. The greater hand grip and hip flexion strength found in black women may be related to their greater muscle mass and known racial differences in body dimensions. No consistent racial differences were observed in the relationship between physical activity and muscle strength, or muscle strength and disability, suggesting that the role of muscle strength in the disablement process does not differ between races. Physical activity and exercise programs may be feasible ways to prevent worsening of disability in blacks and whites. PMID- 9754134 TI - Comparison of cross-sectional and longitudinal designs in the study of aging of upper extremity performance. AB - The purpose of the study was to compare two research designs, namely the cross sectional design and the longitudinal design, in the context of upper extremity performance and age-related changes. Upper extremity performance of 360 randomly recruited healthy, community-dwelling elderly persons was evaluated with reliable and valid sensori-motor tests. Three years later, survivors (n = 264) were reevaluated with the same tests. In many tests, cross-sectional and longitudinal designs were comparable for estimating the changes in upper extremity performance with age. However, in some tests, the decline with age using a cross-sectional design was underestimated. The upper extremity performance decline observed with the longitudinal design was larger than the decline predicted with the cross sectional design. The withdrawal and survivor biases related to the longitudinal design and the cohort bias associated with the cross-sectional design may, in part, explain these results. PMID- 9754135 TI - Decreased axosomatic input to motoneurons and astrogliosis in the spinal cord of aged rats. AB - An increasing body of evidence indicates that aging-related impairments of nervous functions are caused by damage to neuron integrity rather than by loss of neurons. By using electron microscopy, we have examined axosomatic boutons on spinal cord motoneurons derived from aged and young adult Sprague-Dawley rats. The main finding was that about half of the examined motoneuron somata from aged rats had a reduced (50%) bouton coverage, which seemed to be caused by a smaller number of axosomatic bouton profiles. Long stretches of the cell body plasma membrane were apposed by pale processes, and immunolabeling for glial fibrillary acidic protein (GFAP) disclosed that a number of the aged motoneurons appeared embedded in GFAP immunopositive processes. Lumbar motoneurons seemed to be more severely affected than cervical motoneurons. At the ultrastructural level, affected motoneurons disclosed plasma membrane irregularities with appendages/sprout-like extensions that in some cases were sites for axosomatic contacts. PMID- 9754136 TI - Age-specific modulation of light production potential, and alkaline phosphatase and protein tyrosine kinase activities in various age mutants of Caenorhabditis elegans. AB - Previous work has shown that reduction-of-function mutations in the genes daf-2 and age-1 can increase adult life (Age phenotype) of Caenorhabditis elegans and that certain daf-12 alleles considerably amplify this effect in daf-2; daf-12 doubles. We have measured the light production potential (LPP) and alkaline phosphatase (ALP) and protein tyrosine kinase (PTK) activity levels as suitable biochemical markers to further investigate genetic interactions between these genes. The light production assay measures superoxide anion production by freeze thawed worms in assay medium containing sufficient amounts of nicotineamide adenine dinucleotide, reduced form (NADH) and nicotineamide adenine dinucleotide phosphate, reduced form (NADPH) to drive the chemiluminescent reaction at maximal speed, and 5 mM cyanide to fully repress cytosolic superoxide dismutase (SOD). This assay thus provides an estimate of the maximum output of the metabolic pathways involved at the instant of freeze-fixation, and under the condition of the assay. LPP and PTK activities decreased similarly in daf-12(m20), and a control strain that had wild-type alleles of daf-12, age-1, and daf-2. The age dependent decrease of LPP and PTK was reduced in age-1(hx542) and age-1(hx542); daf-2(e1370), and virtually absent in daf-2(e1370) and daf-2(e1370); daf-12(m20) mutant worms. ALP activity increased with age in non-Age genotypes and showed little, if any, age-dependent alteration in daf-2(e1370) and daf-2(e1370); daf 12(m20) mutant worms. Mutation in both age-1 and daf-2 caused no stronger phenotype than a single mutation as estimated by LPP, PTK, and ALP. We propose that (a) daf-2 is the major effector of metabolic activity during adult life, (b) daf-2 downregulates metabolic activity with increasing age, and (c) daf-12 stimulates oxygen consumption independently of daf-2. PMID- 9754137 TI - Comparison of a functional obstacle course with an index of clinical gait and balance and postural sway. AB - BACKGROUND: Older adults commonly experience falls because of balance and mobility problems. Better assessment methods are needed to understand and correct balance and mobility disorders. METHODS: We used a low technology, functional obstacle course (FOC) to measure balance and mobility in 352 community-dwelling elderly participants. To establish concurrent validity of the FOC, we compared performance on the FOC with two established measures of balance and mobility: performance on the Tinetti Index (TI) and postural sway area measured on a force platform. RESULTS: Bivariate correlation analyses revealed significant inverse correlations between FOC completion time, the TI balance and gait subscores, and the TI total score (r = -.73 to -.78). The FOC quality scores and TI balance and subscores gait and TI total scores (r = .76 to .82) were significantly positively correlated. FOC time had significant, but small, positive correlations with sway area with eyes open (r = .18) and closed (r = .17) and nonsignificant correlation with sway area with visual feedback. FOC quality also had significant, but smaller, inverse correlations with sway area with eyes open (r = -.024) and closed (r = -.015), and nonsignificant correlation with sway area with visual feedback. Regression analysis showed that TI gait and balance measures accounted for most of the variance found in FOC performance. CONCLUSIONS: Our findings support the position that the FOC and the TI measure dynamic balance, whereas postural sway measures a different aspect of balance. Advantages of the FOC include the evaluation of environmentally influenced falls and balance problems. PMID- 9754138 TI - Left-truncated data with age as time scale: an alternative for survival analysis in the elderly population. AB - BACKGROUND: The standard approach for survival analysis of the elderly population is to define the survival time as the elapsed time from entry into the study until death, and to adjust by age using stratification and regression procedures. However, the interest is in the study of the aging process and the risk factors related to it, not in the use of time-on-study as the time scale. Here, we present methods to use age as the time scale and compare inferences and interpretations with those obtained using the standard approach. METHODS: A total of 1,315 individuals aged 65 years or older from the city of Barcelona, Spain, were interviewed in 1986 (baseline). The vital status of the cohort was assessed in October 1994. To illustrate the usefulness of age as time scale (alternative approach) instead of time-on-study in the survival analysis of the elderly population, both methods were used to assess the relationship between baseline functional capacity and mortality. RESULTS: Using the alternative approach, we observed that 50% of the sample died at age 80.6 years; this information could not be estimated with the standard approach. Using age as a covariate in the standard analysis with time-on-study as the time scale and using age as the time scale in the alternative analysis, the association of functional capacity at baseline and mortality was of similar magnitude under both analyses. Nevertheless, using the alternative approach, relative risks were slightly lower, and the adjustment by age was tight and was not subject to the inherent assumptions in regression models of the functional relationship of independent variables with outcome. We illustrated the methods with fixed covariates (i.e., gender) and baseline values of time-dependent covariates (i.e., functional capacity), but we discussed the extension of our methods for the analysis of time dependent covariates measured at several visits in a cohort study. Methods proposed here are easily implemented with widely available statistical software packages. CONCLUSIONS: Although the use of standard survival analysis generally produces correct estimates, the use of age as time scale is deemed more appropriate for survival analysis of the elderly: Inferences are easier to interpret and final models are simpler. We therefore recommend the use of age as time scale for survival analysis of the elderly population. PMID- 9754139 TI - Arterial stiffness and hormone replacement use in healthy postmenopausal women. AB - BACKGROUND: Arterial stiffness has been shown to be positively related to cardiovascular disease risk. Little, however, is known about the influence of hormone replacement use on arterial stiffness in females. METHODS: Arterial pulse wave velocity (PWV) and carotid augmentation index (AI, applanation tonometry) were measured in 34 healthy postmenopausal women, including users (n = 18) and nonusers (n = 16) of hormone replacement. RESULTS: There were no significant group differences in any of the physical characteristics including resting blood pressure. None of the measures of arterial stiffness differed between user and nonusers of hormone replacement. CONCLUSIONS: The present cross-sectional study indicates that reduced arterial stiffness does not appear to be a likely mechanism contributing to the lower incidence of cardiovascular disease in postmenopausal women taking hormone replacement. PMID- 9754140 TI - Longevity and gray hair, baldness, facial wrinkles, and arcus senilis in 13,000 men and women: the Copenhagen City Heart Study. AB - BACKGROUND: We have previously reported that men who look older than their contemporaries have a significantly higher risk for myocardial infarction. The purpose of this study was to investigate whether persons with pronounced aging signs such as graying of hair, baldness, or facial wrinkles are prone to a shorter life span compared to their contemporaries. METHODS: In the Copenhagen City Heart Study comprising a random sample of 20,000 men and women, we also recorded, in addition to cardiovascular risk factors, data on signs of aging: extent of gray hair, baldness, facial wrinkles, and arcus senilis (corneal arcus). During 16 years of follow-up, 3,939 persons (1,656 women and 2,283 men) had died. The Cox regression model for proportional hazards, which included age as an explanatory variable, was used for descriptive analysis of the correlation between these aging signs and all-cause mortality. RESULTS: We found no correlation between the mortality and the extent of graying of the hair, or baldness or facial wrinkles in either of the sexes, irrespective of age. A single exception was observed in a small subgroup of men with no gray hair. They had a slightly, but significantly, lower mortality than the rest [relative risk (RR) = .81, 95% confidence interval (CI) .67-.98; p < .05]. The presence of arcus senilis was significantly correlated with a shorter life span in women (RR = 1.25, 95% CI 1.08-1.46; p < .01). For men the same tendency was found, but the correlation was not statistically significant. CONCLUSION: We conclude that the degrees of graying of the hair, baldness, and facial wrinkles are not predictive of a shorter life span in men and women in the Copenhagen City Heart Study. PMID- 9754141 TI - Symptom severity of osteoarthritis of the knee: a patient-based measure developed in the veterans health study. AB - BACKGROUND: Our objective was to develop a patient-based measure of the severity of osteoarthritis of the knee, focusing on symptomatology, that may be used in conjunction with measures of health-related quality of life in monitoring the health status of outpatients. METHODS: We surveyed a random sample of male outpatients at Boston-area Veterans Affairs medical centers who were identified as having osteoarthritis of the knee according to a three-question screen. Structured interviews included 12 items covering five domains of symptoms (global severity, 4 items; pain with activity, 3 items; pain at rest, 2 items; impaired mobility, 2 items; and sensations of crepitus, 1 item), which were derived from clinical texts, consensus statements, and previously developed severity indices. Interviews also included a detailed medical history. Health-related quality of life was measured by the Medical Outcomes Study Health Status Survey (SF-36). Factor analysis and evaluation of multiattribute scales were used to evaluate the structural relationships between and within the five domains of symptoms. RESULTS: We identified 415 of the 1770 screened outpatients as having osteoarthritis of the knee. Internal consistencies of the five domains ranged from .50 to .72, with substantial convergence between domains. The 12 items comprise a summary index with high internal consistency (alpha = .88). Overall severity, defined as the mean of the 12 items after standardization, was moderately correlated with the SF-36 component summaries: r = -.48 for physical; r = -.30 for mental. CONCLUSIONS: Our measure provides a reliable index that represents symptomatic severity of osteoarthritis of the knee, which may be useful in comparing patient groups and assessing health outcomes; subscales may help characterize temporal changes, including responses to treatment. PMID- 9754142 TI - Disaggregating pain and its effect on physical functional limitations. AB - BACKGROUND: Pain is a common impairment that limits the abilities of older persons. The purposes of this article are to: (i) describe the distribution of pain location using the McGill Pain Map (MPM) in a community-based cohort of aged subjects; (ii) investigate whether individual areas of pain could be sensibly grouped into regions of pain; (iii) determine whether intensity, frequency, and location constitute independent dimensions of pain; and (iv) determine whether these three pain dimensions make differential contributions to the presence of self-reported physical functional limitations. METHODS: A total of 833 Mexican American and European American subjects, aged 65-79 years, were enrolled in the San Antonio Longitudinal Study of Aging and were interviewed in their homes between 1992 and 1996. A total of 373 (46%) of the subjects reported having pain in the past week. Physical functional limitations were ascertained using the nine items from the Nagi scale. Three composite scales were created: upper extremity, lower extremity, and total. Pain intensity and frequency were ascertained using the McGill Pain Questionnaire. Pain location was ascertained by using the MPM. RESULTS: Pain was reported in every area of the MPM. Using multiple groups confirmatory factor analysis, the 36 areas were grouped into 7 regions of pain: head, arms, hands and wrists, trunk, back, upper leg, and lower leg. Among persons with pain, pain frequency, intensity, and location were weakly associated with each other. Pain regions were primarily independent of each other, yet weak associations existed between 6 of the 21 pair-wise correlations between regions. Pain regions were differentially associated with individual physical functional limitations. Pain in the upper leg was associated with 8 of the 9 physical tasks. In multivariate analyses, age, gender, and ethnic group accounted for only 2-3% of the variance in physical tasks. In multivariate analyses, age, gender, and ethnic group accounted for only 2-3% of the variance in physical functional limitations. Pain intensity accounted for 5-6% of the variance in the composite scores of functional limitation. Pain frequency accounted for 4-5% of the variance in upper extremity limitations but did not contribute to the modeling of lower extremity limitations. In contrast, pain location accounted for 9-14% of the variance in physical functional limitations. CONCLUSIONS: We tested a method for ascertaining pain location and clearly demonstrated that pain location is an important determinant of self-reported physical functional limitations. The MPM methodology may be used in population-based studies or in clinical samples that focus on specific impairments and seek to control for pain frequency and intensity. Future studies can link specific diseases with the common impairment of pain and tease out the pathways that lead to other impairments (e.g., weakness), functional limitations, and disability. PMID- 9754143 TI - Does comorbid disease interact with cancer? An epidemiologic analysis of mortality in a cohort of elderly breast cancer patients. AB - BACKGROUND: Although widely believed that co-occurring chronic diseases in elderly persons do not act independently in causing death, there has been little empirical research assessing prognostic interrelationships between comorbidities. METHODS: Nonconcurrent prospective follow-up of 3,549 Virginia-resident elderly women diagnosed with a first breast cancer and 2,114 elderly women with no breast cancer history admitted to Virginia hospitals with principal diagnoses of genital prolapse during 1986-1988 was conducted through linkage of cancer registry and Medicare administrative records. Aggregate comorbidity was measured from Medicare claims via the Charlson comorbidity index (CCI). Mortality rates and relative risks were estimated for the breast cancer and non-breast-cancer groups stratified by the presence and level of comorbidity. Proportional hazards models were used to estimate Rothman's synergy index (S) measure of additive interaction. RESULTS: Over full follow-up, the excess mortality rate for women with breast cancer and other comorbidity was 17% greater than expected under the null hypothesis that risks were additive and independent (S = 1.17, p = .12). Stratified analyses revealed a pattern of S estimates across cancer stage subgroups that was biologically sensible, but this pattern was not supported by strong statistical evidence. CONCLUSIONS: This study provides the first empirical estimates of statistical interaction between breast cancer and other chronic comorbidity. S index values tended to be small, but these small effects would translate into substantial numbers of deaths attributable to interaction between cancer and comorbidity. Interactions between breast cancer and comorbid disease should be explored further in large studies that can estimate these effects with increased precision. PMID- 9754144 TI - Soliciting defined populations to recruit samples of high-risk older adults. AB - BACKGROUND: Generalizable research on high-risk older persons requires samples that are both large enough for adequate statistical power and similar enough to community populations that its results can be generalized to them. We tested the effectiveness and efficiency of mixed-mode (mail-telephone) solicitation of a defined population as a method for recruiting a large, representative sample for a randomized trial of outpatient geriatric evaluation and management (GEM). METHODS: Fee-for-service, community-dwelling older Medicare beneficiaries were mailed a short self-administered screening questionnaire. Eligible respondents were called to assess eligibility and willingness to give consent; consenters were called again for baseline data. Information about nonrespondents, ineligibles, and refusers was obtained from the Health Care Financing Administration. RESULTS: The response rate to the screening questionnaire was 61.1%. Of the respondents, 13.2% were eligible for the study and, of those, 34.4% agreed to participate. Response rates appeared to be influenced by small financial incentives and by subjects' age, race, sex, location of residence, and use of hospitals in the previous year. Consent rates were influenced by age and sex. The final sample (N = 522) was representative of community high-risk respondents in racial composition, previous use of hospitals, and probability of repeated admission (Pre) in the future, but it was slightly younger and contained a higher percentage of men. Recruitment costs averaged $286.92 per consenting person. CONCLUSIONS: Mixed-mode solicitation of defined populations can produce, at reasonable cost, large samples whose representativeness of community high-risk populations can be determined. Procedures that may enhance the success of this approach include: advance communication with members of the target population and their families and physicians; provision of medical and small financial incentives; continuous monitoring of recruitment results; and attention to subjects' needs for convenience, time, transportation, and reassurance. PMID- 9754145 TI - Effects of dehydroepiandrosterone replacement in elderly men on event-related potentials, memory, and well-being. AB - BACKGROUND: In humans, concentrations of the adrenal steroid hormone dehydroepiandrosterone (DHEA) and its sulfate ester (DHEAS) decline with age. Results from studies in rodents have suggested that DHEA administration can improve memory performance as well as neuronal plasticity. However, a first study from our laboratory could not demonstrate beneficial effects of DHEA substitution on cognitive performance and well-being in elderly subjects. To further evaluate whether DHEA replacement has effects on the central nervous system, an experiment using event-related potentials (ERPs) was conducted. METHODS: In this placebo controlled crossover study, 17 elderly men (mean age, 71.1 +/- 1.7 yr; range 59 81 yr) took placebo or DHEA (50 mg/day) for 2 weeks (double blind). After each treatment period subjects participated in an auditory oddball paradigm with three oddball blocks. In the first two blocks subjects had to count the rare tone silently, whereas, in the third block they had to press a button. In addition, memory tests assessing visual, spatial, and semantic memory as well as questionnaires on psychological and physical well-being were presented. RESULTS: Baseline DHEAS levels were lower compared with young adults. After 2-week DHEA replacement, DHEAS levels rose 5-fold to levels observed in young men. DHEA substitution modulated the P3 component of the ERPs, which reflects information updating in short-term memory. P3 amplitude was increased after DHEA administration, and only selectively in the second oddball block. DHEA did not influence P3 latency. Moreover, DHEA did not enhance memory or mood. CONCLUSIONS: A 2-week DHEA replacement in elderly men results in changes in electrophysiological indices of central nervous system stimulus processing if the task is performed repeatedly. However, these effects do not appear to be strong enough to improve memory or mood. PMID- 9754146 TI - Periodic leg movements during sleep and sleep disturbances in elders. AB - BACKGROUND: Periodic limb movements in sleep (PLMS) are an increasingly pervasive disturbance for aging adults. The aims of this experiment were: (a) to describe the index of periodic limb movements in sleep (myoclonus index [MI] in elderly subjects with complaints of poor sleep or depression (N = 22; 68 +/- 5.5 SD years); and (b) to correlate MI with sleep history, depression scores, and objective and subjective indices of sleep. METHOD: Sleep and leg movements were assessed for 5 consecutive nights. Between-subjects, nonparametric correlations were examined between mean MI and sleep history, depression scores, and objective and subjective sleep characteristics. Associations among within-subject night-to night variabilities of MI, objective, and subjective variables were examined with repeated measures ANCOVA, entering MI as a covariate. RESULTS: A remarkably high level of MI was found (median 25.8 events per hour; 86% of subjects > 5). Nevertheless, no associations were found between MI and sleep disturbance measures. CONCLUSION: These results extend previous reports that PLMS are remarkably persuasive in elderly volunteers and support other reports questioning whether there is a distinct PLMS syndrome. PMID- 9754147 TI - Interrelationships among disablement concepts. AB - BACKGROUND: Understanding interrelationships among disablement concepts is critical to the design of future disability treatment and prevention interventions. METHODS: This study uses cross-sectional data to examine the relationships among physiologic impairments, functional limitations, and disability in a moderately disabled sample of 207 community-dwelling older adults. RESULTS: As hypothesized, the data revealed statistically significant curvilinear relationships of upper and lower extremity strength and balance with mobility in this older sample. Multivariate analyses further clarified the hypothesized causal mechanism among the disablement concepts by demonstrating that most of the association of muscle strength and balance with disability was through the intermediary role of mobility limitations. CONCLUSIONS: The findings from this study highlight the value of clinical trials that focus on prevention or treatment of mobility limitations as a means of preventing disability; our findings underscore the need for future research that examines the effects of other variables believed to influence disablement in late life. PMID- 9754148 TI - Previous disability as a predictor of outcome in a geriatric rehabilitation unit. AB - BACKGROUND: Functional status at admission has been shown consistently to predict rehabilitation results, but the impact of previous disability has been seldom considered. METHODS: A prospective follow-up study of elderly patients admitted to a geriatric rehabilitation unit in Madrid, Spain, was carried out. The study population comprised 135 subjects aged 65 years or older, who were consecutively admitted during a 7-month period. Outcome variables included the Barthel Index (BI) at discharge, the improvement in BI from admission to discharge, the achieved percentage of potential gain, and the efficiency of gains. Previous BI, admission BI, diagnosis, source (hospital/others), mental status, age, and gender were examined as explanatory variables. RESULTS: In multiple regression analysis, previous BI was the only significant independent predictor for all the outcome variables. For each 5-point increase in previous BI, the increase in BI at discharge was 1.7 (p = .007). Corresponding values for the achieved percentage of potential gain and for the efficiency of gains were 0.05 (p = .01) and 0.05 (p = .04), respectively. Except for the achieved percentage of potential gain, admission BI and source of referral were also independent significant predictors of outcome. CONCLUSIONS: Previous functional situation of elderly people is important to predict rehabilitation outcome, even after taking into account information on disability at admission. As a consequence, a measure of the achieved percentage of potential gain corrected by the preadmission functional status is proposed, especially in the case of elderly patients. PMID- 9754149 TI - Refractive changes after lens implantation in childhood. PMID- 9754150 TI - Clinical and radiologic lacrimal testing in patients with epiphora. PMID- 9754151 TI - Automated perimetry and threats to fixation. PMID- 9754152 TI - Automated perimetry and threats to fixation. PMID- 9754153 TI - Retrobulbar versus sub-Tenon's corticosteroid therapy. PMID- 9754154 TI - Retrobulbar versus sub-Tenon's corticosteroid therapy. PMID- 9754155 TI - Laser treatment of macular diseases. PMID- 9754156 TI - Visual acuity outcomes with appositional suprachoroidal hemorrhage. PMID- 9754157 TI - Combination of systemic acetazolamide and topical dorzolamide. PMID- 9754158 TI - Floppy eyelid syndrome. PMID- 9754159 TI - Angioarchitectural changes in branch retinal vein occlusion. PMID- 9754160 TI - Topographic detection of photorefractive keratectomy. PMID- 9754161 TI - Age-related macular degeneration: is help on the way? PMID- 9754162 TI - Silicone oil in the repair of complex retinal detachments. A prospective observational multicenter study. AB - OBJECTIVE: This study aimed to report anatomic and visual acuity outcomes and complications after 1000-centistoke silicone oil was used as a retinal tamponade for the treatment of complex retinal detachments. DESIGN: Prospective observational multicenter study conducted at community and university-based ophthalmology clinics. PARTICIPANTS: The study cohort consisted of 2439 patients (2573 eyes) treated for complex retinal detachments associated with cytomegalovirus (CMV) necrotizing retinitis or a non-CMV etiology, including proliferative diabetic retinopathy, giant retinal tears, proliferative vitreoretinopathy, or ocular trauma. INTERVENTION: Vitrectomy surgery was performed for complex retinal detachment with 1000-centistoke silicone oil as the retinal tamponade. MAIN OUTCOME MEASURES: Anatomic outcomes were complete retinal attachment and macular attachment. Visual acuity outcomes were ambulatory vision (> or = 4/200) and preservation of preoperative visual acuity. Complications were rates of secondary intraocular pressure elevation (> or = 30 mmHg), hypotony (< or = 5 mmHg), corneal opacification (including band keratopathy, corneal edema, and corneal abrasions), oil emulsification, and cataract. Outcomes were assessed 6, 12, and 24 months after surgery. RESULTS: At the 6-month examination, the retina was completely attached in 178 (78%) of 228 CMV eyes and in 855 (70%) of 1219 non-CMV eyes. The macula was attached in 216 (95%) of 228 and 1062 (89%) of 1189 CMV and non-CMV eyes, respectively. Ambulatory vision was noted in 151 (65%) of 234 CMV eyes and in 480 (38%) of 1251 non-CMV eyes. Visual acuity was preserved in 106 (46%) of 230 and 1035 (84%) of 1229 CMV and non-CMV eyes, respectively. The corresponding rates of complications for CMV and non-CMV eyes were: elevated intraocular pressure, 0 (0%) of 196 and 35 (3%) of 1196; hypotony, 11 (6%) of 196 and 228 (19%) of 1196; corneal opacity, 13 (6%) of 229 and 326 (26%) of 1248; emulsification, 3 (1%) of 211 and 29 (3%) of 959; and cataract in phakic eyes, 118 (64%) of 185 and 50 (63%) of 80. CONCLUSIONS: Retinal reattachment was achieved in the majority of eyes using vitrectomy and silicone oil retinal tamponade. Complication rates generally were less frequent in CMV eyes, but follow-up was shorter in this group of patients, largely because of reduced life expectancy. Cataract frequently developed in phakic eyes of study patients. Use of 1000-centistoke silicone oil can be considered in the management of complex retinal detachments associated with multiple etiologies. PMID- 9754163 TI - Surgical removal of extensive peripapillary choroidal neovascularization associated with presumed ocular histoplasmosis syndrome. AB - OBJECTIVE: This study aimed to report the visual outcome of surgical removal of extensive peripapillary choroidal neovascularization (CNV) due to presumed ocular histoplasmosis syndrome (POHS). DESIGN: Retrospective review of the records of all patients seen at the Barnes Retina Institute who underwent surgical removal of extensive peripapillary CNV associated with POHS and who had at least 12 months of follow-up. PARTICIPANTS: Seventeen consecutive eyes (in 14 patients) undergoing surgical removal of extensive peripapillary CNV associated with POHS were studied. INTERVENTION: Pars plana vitrectomy and surgical removal of CNV were performed. MAIN OUTCOME MEASUREMENTS: Best-corrected Snellen visual acuity, funduscopic examination, and intravenous fluorescein angiography were obtained before surgery and at regular intervals after surgery. RESULTS: In 14 of 17 eyes, the peripapillary CNV was subfoveal, and in 3 eyes, it was extrafoveal. All three eyes with extrafoveal CNV were not eligible for laser treatment according to Macular Photocoagulation Study guidelines because treatment would have spared less than 1.5 contiguous clock-hours of retina temporal to the optic disc. Follow up ranged from 17 to 57 months, with a median of 32 months. In eyes with subfoveal CNV, best-corrected preoperative Snellen visual acuity ranged from 20/25 to counting fingers at 2 feet with a median of 20/200, and best-corrected final Snellen visual acuity ranged from 20/25 to 20/200 with a median of 20/40. In 7 (50%) of 14 eyes, a final Snellen acuity of 20/40 or better was achieved, and in all cases except 1, visual acuity improved or did not change with surgery. In the three eyes with extrafoveal CNV, best-corrected preoperative Snellen visual acuity ranged from 20/20 to 20/400 with a median of 20/200, and best corrected final Snellen visual acuity was 20/20 in all cases. In addition, visual acuity improved with surgery. CONCLUSIONS: The data from this small retrospective study suggest that surgical removal may provide visual benefit in selected cases of extensive peripapillary CNV due to POHS. PMID- 9754164 TI - Surgical removal of subfoveal iatrogenic choroidal neovascular membranes. AB - OBJECTIVE: To present the cases of two patients with laser-induced iatrogenic subfoveal choroidal neovascular membranes (CNVMs) who underwent surgical removal of the membranes with favorable outcomes. DESIGN: Interventional case reports. PARTICIPANTS: Two patients with iatrogenic subfoveal CNVM. One case developed after laser treatment for macular edema due to branch retinal vein occlusion, and the second case developed after focal laser photocoagulation for diabetic retinopathy. INTERVENTION: Surgical removal by pars plana vitrectomy. MAIN OUTCOME MEASURES: Visual acuity, scotoma, retinal examination with fundus photography, and fluorescein angiography before surgery and during the postoperative period. RESULTS: Both patients underwent surgical removal after progression of the membrane with severe visual loss of 20/200 was noted. At present follow-up, there is a significant improvement in visual acuity and a reduction in the size of the scotoma. No recurrence of CNVM is noted. CONCLUSION: Both patients with laser-induced iatrogenic subfoveal CNVM achieved a good visual outcome after surgical removal of the membrane. The reasons for a good surgical result are thought to be twofold. First, the origin of the CNVM is extrafoveal at the site of laser application. The chance for foveal cone cell damage during the surgery is reduced. Second, the degree of cellular destruction in iatrogenic CNVM is usually focal without extensive retinal photoreceptor cell and retinal pigment epithelial damage. Therefore, a better chance of postoperative visual recovery is anticipated. PMID- 9754165 TI - Visual field defect after pars plana vitrectomy. AB - OBJECTIVE: This study aimed to report the occurrence of visual field defects after pars plana vitrectomy (PPV) for the treatment of each of the following conditions: macular hole (MH), subretinal neovascular membrane (SRNVM), and epiretinal membrane proliferation (EMP). This study also aimed to speculate on the pathogenic mechanisms for the observed field defects. DESIGN: Noncomparative case series. PARTICIPANTS: The study included 48 subjects (50 eyes). Twenty-one of the 50 eyes had stage III MH, 13 eyes had SRNVM, and 16 eyes had EMP. TESTING: Goldmann kinetic perimetry was performed postoperatively. MAIN OUTCOME MEASURE: Visual field defects. RESULTS: Nine (18%) of the 50 eyes had visual field defects. Four (19%) of the 21 eyes with MH and 5 (38%) of the 13 eyes with SRNVM had visual field defects. Of the 16 patients who had epiretinal membrane peeling, none had a visual field defect. An air-fluid exchange had been performed in all patients found to have a postvitrectomy field defect. The difference in rate of visual field defects in eyes that had air-fluid exchange (EMP group) was statistically significant (P < 0.05, chi-square). No significant correlation was found between visual field defect and preoperative intraocular pressure, postoperative intraocular pressure, patient's age, and iatrogenic detachment of the vitreous cortex. The field defects identified were altitudinal (2 eyes), baring of the blind spot (1 eye), inferotemporal (3 eyes), inferonasal (2 eyes), and superonasal (1 eye). CONCLUSIONS: Central and peripheral visual field defects may occur after PPV for the treatment of MHs or SRNVMs. Air-fluid exchange procedure was the common denominator in all of the patients found to have visual field deficit. The etiology is likely to be trauma to the optic nerve region during the air-fluid exchange procedure. PMID- 9754166 TI - Plasmin enzyme-assisted vitrectomy in traumatic pediatric macular holes. AB - OBJECTIVE: This study aimed to evaluate the benefit of plasmin enzyme-assisted macular hole surgery on a consecutive series of pediatric patients with traumatic macular holes. DESIGN: Prospective noncomparative case series operated on at William Beaumont Hospital between July 13, 1996, and November 16, 1996, and observed for at least 6 months. PARTICIPANTS: During this interval, the authors operated on four eyes from four consecutive patients who were 14 years of age or younger with traumatic macular holes. INTERVENTION: The patients underwent plasmin enzyme-assisted pars plana vitrectomy with membrane peeling, fluid-gas exchange, and postoperative positioning. The enzyme used was 0.4 international unit (IU) of autologous plasmin enzyme. MAIN OUTCOME MEASURES: Snellen lines of improvement in visual acuity and rate of final visual acuity of 20/40 or greater, and incidence of complications and reoperations were measured. RESULTS: All four macular holes were closed successfully. Follow-up was from 6 to 12 months. There were no reoperations. Visual acuity improved from four to eight lines in all eyes. Three eyes (75%) achieved a postoperative visual acuity of 20/40 or better. Three eyes (75%) had transient, posterior, subcapsular cataracts develop: two of the eyes after surgery and one as a result of the initial injury. CONCLUSION: The treatment of pediatric traumatic macular holes with plasmin enzyme-assisted vitrectomy, membrane peeling, and gas-fluid exchange resulted in closure of the macular holes with significant visual improvement. PMID- 9754167 TI - The incidence of ophthalmologic interventions in children with birth weights less than 1251 grams. Results through 5 1/2 years. Cryotherapy for Retinopathy of Prematurity Cooperative Group. AB - OBJECTIVE: This study aimed to report the frequency of ophthalmologic surgical and medical therapies provided to children with birth weights less than 1251 g who had all stages of retinopathy of prematurity (ROP). In addition, this study aimed to report the initial age at which such procedures are provided and to report the frequency of cerebrospinal fluid shunts. DESIGN: Observational case series with prospective data collection. PARTICIPANTS: Children from the Multicenter Trial of Cryotherapy for Retinopathy of Prematurity (CRYO-ROP) with birth weights less than 1251 g served as subjects. Group A included 257 children from all 23 CRYO-ROP study centers who had threshold ROP, who had participated in the randomized trial of cryotherapy, and who had survived to age 1 year. Group B included 1208 children from 5 of the 23 study centers who had varying severity of ROP (69 had threshold ROP) and who had participated in a 5 1/2-year study of the natural history of ROP. MAIN OUTCOME MEASURES: Investigators documented medical and surgical ophthalmologic interventions through age 5 1/2 years as well as cerebrospinal fluid shunting surgery for hydrocephalus through age 2 years. RESULTS: Group A was composed of 257 children with threshold ROP who underwent 226 ocular interventions in addition to cryotherapy (0.9 intervention per child). The most common treatments performed on the randomized cohort of children were vitrectomy (26% of patients), lensectomy (18%), amblyopia therapy (20%), and strabismus surgery (10%). Cataract surgery not associated with vitrectomy was performed infrequently (2%) and was performed equally often in treated and control eyes. Amblyopia therapy was prescribed as often for treated as for control eyes. Cerebrospinal fluid shunts were placed in 11% of these children. Group B was composed of 1208 natural history patients who underwent 239 ophthalmologic interventions (0.4 intervention per child). Strabismus surgery was the most commonly performed procedure for the natural history cohort of children (6% of the children). Amblyopia therapy was prescribed for 7% of the natural history patients. Cerebrospinal fluid shunts were required by 3% of the natural history infants, more often in children with more severe ROP. CONCLUSIONS: These premature infants underwent a large number of ophthalmologic treatments during the first 5 1/2 years of life. The long-term costs of both extreme prematurity and ROP include not only the initial ablative therapy for ROP and societal loss due to blindness that still occurs in some cases, but also the ongoing costs of caring for eye problems. PMID- 9754168 TI - A comparison of laser photocoagulation with trans-scleral cryotherapy in the treatment of threshold retinopathy of prematurity. AB - OBJECTIVE: The goal of this study was to determine whether there was a significant difference between the visual outcomes of eyes with threshold retinopathy of prematurity (ROP) treated with trans-scleral cryotherapy compared to those treated with laser photocoagulation. DESIGN: Extended follow-up study of a prospective, randomized clinical trial. PARTICIPANTS: Fifty-two patients with bilateral threshold ROP participated. Follow-up data were available for 25 of these patients. INTERVENTION: Patients were randomized to receive cryotherapy in one eye and laser photocoagulation in the other eye. MAIN OUTCOME MEASURES: The best-corrected visual acuity of each eye was measured. Best-corrected visual acuities of 20/50 or better were classified as "good" clinical outcomes, whereas those 20/60 or worse were considered "poor" outcomes. A secondary outcome of this study was the spherical equivalent (SE) of each eye's most recent refraction. RESULT: At an average follow-up point of 5.8 years (range, 4.3-7.6 years), the odds that an eye treated with laser had a good clinical outcome were 6.91 times greater than for eyes treated with cryotherapy (95% confidence interval, 1.70 28.0; n = 21). Additionally, the laser-treated eyes were less myopic with a mean SE of-3.05 diopters compared to a mean SE of -5.08 diopters for the cryotherapy treated eyes (P = 0.0072, n = 23). CONCLUSION: The authors' study suggests that laser photocoagulation for threshold ROP was more likely to result in a good clinical outcome with better final visual acuity and less myopia compared to cryotherapy treatment. PMID- 9754169 TI - Indocyanine-green videoangiography of drusen as a possible predictive indicator of exudative maculopathy. AB - OBJECTIVE: Recent studies have shown that indocyanine-green videoangiography (ICG V) is useful to image occult choroidal neovascularization. The authors studied the ICG-V findings in fellow drusen eyes of patients with unilateral exudative age-related macular degeneration (AMD). The authors also studied the occurrence of exudative changes to determine whether ICG-V is useful in predicting future exudative changes in these eyes with only drusen. DESIGN: Cohort study. PARTICIPANTS: The authors studied 432 consecutive patients diagnosed with unilateral exudative AMD in whom the fellow eye had only drusen by clinical fundus examination and fluorescein angiography. All of these eyes had ICG-V performed. Follow-up data were obtained in all eyes with abnormal indocyanine green (ICG) angiograms and randomly sampled ICG angiograms of normal eyes. MAIN OUTCOME MEASURES: The initial ICG findings were classified as showing normal or abnormal hyperfluorescence. Abnormal hyperfluorescence eyes were subdivided into focal spots (focal areas of hyperfluorescence < 1 disc area in size) and plaques (areas of hyperfluorescence > 1 disc area). The development of exudative changes in eyes with normal and abnormal hyperfluorescence was compared. RESULTS: Of the 432 fellow eyes, 386 (89%) eyes with drusen had a normal ICG-V study, whereas 46 (10 focal spots and 36 plaques) (11%) eyes had an abnormal ICG-V. Exudative changes occurred in 6 (10%) of 58 normal ICG eyes and 9 (24%) of 38 eyes with abnormal ICG findings during a mean follow-up period of 21.7 months. The difference between drusen eyes with normal ICG angiograms and those with plaques on ICG-V regarding future exudative changes (10% vs. 27%, respectively) was statistically significant (P = 0.038). CONCLUSIONS: Abnormal ICG findings were found in 11% of eyes with clinically and fluorescein angiographically nonsuspicious drusen. The subgroup of patients with plaques on ICG-V had a higher chance of having exudative changes develop. Indocyanine-green videoangiography may be a predictive indicator of future exudative changes in eyes with drusen. A much larger prospective study seems justified. PMID- 9754170 TI - A practical diagnostic test for choroideremia. AB - OBJECTIVE: This study aimed to establish a practical diagnostic test for choroideremia (CHM) and to show its application in a family with the clinical diagnosis of choroideremia. DESIGN: Case series. PARTICIPANTS: Sixteen males from 13 families with clinically documented CHM and unaffected normal males were enrolled in this study. METHODS: Protein extracted from either leukocytes or Epstein-Barr virus-transformed lymphocytes was subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Immunoblot analysis of the protein was performed with two monoclonal antibodies, one against the CHM gene product, Rab escort protein-1 (REP-1), and the other against the alpha-subunit of farnesyl transferase. DNA was extracted from peripheral leukocytes and subjected to polymerase chain reaction-single stranded conformation polymorphism analysis using primers for the exons of the CHM gene. Where altered mobility of the DNA fragments was detected, direct sequencing of that exon was compared with the published normal sequence. RESULTS: The authors detected REP-1 in protein samples extracted from lymphoblastoid cell lines from female carriers but not from CHM males. The authors also showed the absence of REP-1 in the peripheral leukocytes of males affected with CHM. In one male who lacked REP-1, direct sequencing of exon 7 showed a cytosine-to-thymine transition mutation (Arg293X) in the CHM gene. CONCLUSIONS: The clinical diagnosis of CHM can be confirmed simply by immunoblot analysis with anti-REP-1 antibody, showing the absence of REP-1 protein in peripheral blood samples. Because all known mutations in the CHM gene create stop codons that truncate the protein product and result in absence of REP 1, the authors predict that most patients with CHM can be diagnosed by this procedure. PMID- 9754171 TI - Intraocular light scatter in patients with choroideremia. AB - OBJECTIVE: This study aimed to evaluate the extent of intraocular light scatter in patients with choroideremia. DESIGN: Prospective case-control study. PARTICIPANTS: Twelve male patients with choroideremia who had predominantly minimal or no posterior subcapsular cataract (PSC) lens opacities and visual acuities of 20/40 or better and 30 age-similar control subjects with normal vision and no lens opacities were studied. INTERVENTION: Intraocular light scatter was measured using a van den Berg Straylightmeter. MAIN OUTCOME MEASURES: Visual acuities, letter contrast sensitivities, Goldmann visual fields using a II4e target, and straylight parameters were obtained for each patient. Lenses were assessed by slit-lamp biomicroscopy to determine whether there were PSC opacities. The degree of retinal pigment epithelial and choroidal degenerative changes was evaluated from color fundus photographs. RESULTS: Three of the patients with choroideremia who had clinically apparent PSC lens opacities showed an increase in intraocular light scatter. More notable was the fact that seven of the remaining nine patients who did not have any clinically apparent changes in the lens also had a considerable increase in the intraocular light scatter as compared to the control subjects. The relative elevations of the log straylight parameters of the patients with choroideremia, as compared to age-similar control subjects, were correlated significantly with their log visual field areas (r = 0.69, P < 0.05). CONCLUSIONS: Intraocular light scatter may be increased in patients with choroideremia, even in the absence of clinically observable PSC opacities. It is hypothesized that the increase in light scatter may be caused by changes in the posterior subcapsular region of the lens before the formation of frank PSC cataracts. The increased straylight could, at least in part, account for the disability glare reported by these patients. PMID- 9754172 TI - Outcomes in anterior uveitis associated with the HLA-B27 haplotype. AB - OBJECTIVE: This study aimed to test the hypothesis that patients presenting with anterior uveitis who are HLA-B27 positive, either with or without associated systemic disease, have a less-favorable outcome than do patients with idiopathic anterior uveitis who are HLA-B27 negative. DESIGN: Retrospective case-controlled series. PARTICIPANTS: Ninety-seven patients who were HLA-B27 positive with no systemic disease, 94 patients who were HLA-B27 positive with systemic disease, and 72 patients who were HLA-B27 negative who presented with anterior uveitis were studied. MAIN OUTCOME MEASURES: Ocular complications (e.g., secondary glaucoma, cataract formation, pupillary synechiae, vitritis, cystoid macular edema, and optic disc edema), medical and surgical treatment, number of recurrent attacks, and final visual acuity were recorded for all patients. RESULTS: The patients who were HLA-B27 positive, either with or without systemic disease, experienced a greater number of complications than did the patients who were HLA B27 negative. Periocular corticosteroids, systemic corticosteroids, and systemic immunosuppressive chemotherapy were required in a far greater number of HLA-B27 positive patients than in HLA-B27-negative patients (60% vs. 11%, 53% vs. 7%, and 18% vs. 1%, respectively; P < 0.001). The percentage of legally blind eyes was significantly greater in the HLA-B27-positive group, both with and without systemic disease, when compared with the HLA-B27-negative group (11% vs. 2%; P < 0.005). CONCLUSIONS: The prognosis of anterior uveitis associated with the HLA B27 haplotype, either with or without associated systemic disease, is less favorable when compared with that of HLA-B27-negative patients with idiopathic anterior uveitis. PMID- 9754174 TI - High levels of interleukin-12 in the aqueous humor and vitreous of patients with uveitis. AB - OBJECTIVE: This study aimed to investigate the role of interleukin-12 (IL-12) and interleukin-10 (IL-10) in initiation and maintenance of intraocular inflammation. DESIGN: Case series. PARTICIPANTS: Aqueous humor and vitreous levels of IL-12 and IL-10 were measured in 22 patients with uveitis undergoing cataract surgery, paracentesis of the anterior chamber, and/or vitrectomy for diagnostic reasons, and in 4 patients with cataract only. INTERVENTION: Aqueous humor and vitreous levels of IL-12 and IL-10 were measured with specific enzyme-linked immunosorbent assays. MAIN OUTCOME MEASURES: Disease activity was correlated to IL-12 levels in the aqueous humor and the vitreous of patients with uveitis. RESULTS: Cytokine levels found in the anterior chamber and the vitreous are presented in picogram/milliliter (medium; range). The highest IL-12 levels were found in patients with active uveitis (108.5 pg/ml; 72-293 pg/ml). Interleukin-12 in patients with moderate uveitis or with their disease in remission was lower (32 pg/ml; 15-94 pg/ml) than in patients with active disease (P > 0.001) but higher than in the control group (10.5 pg/ml; 9-14 pg/ml). Interleukin-10 was detectable in only 3 of 22 patients with uveitis (12 pg/ml; 9-23 pg/ml). CONCLUSION: The authors found statistically significant differences of IL-12 levels in the various patient groups (active vs. inactive vs. control). These results support the idea that these uveitis cases represent type 1 (Th1)-T lymphocyte-mediated diseases in which IL-12 plays a pivotal role in the initiation and maintenance of the intraocular inflammation. The high levels of IL-12 in the vitreous and/or aqueous humor of the patients with uveitis suggest that susceptibility or resistance to ocular autoimmunity may be connected to a genetic predisposition to an elevated Th1 response. PMID- 9754173 TI - Nontuberculous mycobacterial keratitis in south Florida. AB - OBJECTIVE: This study aimed to review the clinical features, therapeutic response, and histopathology of cases of nontuberculous mycobacterial keratitis at the Bascom Palmer Eye Institute. DESIGN AND PARTICIPANTS: Retrospective review of medical records, clinical photographs, histopathology, and microbiology of 24 cases of nontuberculous acid-fast keratitis over the past 15 years. RESULTS: Causal organisms included Mycobacterium chelonae (16), M. fortuitum (3), M. avium intracellulare (2), M. nonchromogenicum (1), M. triviale (1), and M. asiaticum (1). Clinically, the keratitis had a superficial location except in those patients with a history of surgery. Amikacin was the most commonly used antibiotic (63%). Three patients were treated with Clarithromycin. In one patient, it was stopped because of toxicity; the other two had resolution of their infiltrates. Fifty-five percent did not respond to topical antimicrobial therapy. The organisms as a group were sensitive to amikacin and Clarithromycin and resistant to the fluoroquinolones. Sixty-four percent of the group that failed to respond to medical treatment were treated with steroids after the diagnosis was known, in comparison to 44% of the group treated successfully with medications. The histopathology of the patients treated with steroids showed minimal inflammation despite a large number of organisms, in contrast to the dense infiltrates seen in the specimens from patients not treated with topical steroids. CONCLUSION: Nontuberculous mycobacterial keratitis is a chronic insidious infection that is often unresponsive to medical therapy. The authors recommend that steroids be withheld. Based on the authors' experience of three patients, topical Clarithromycin may hold promise as a therapeutic agent. Lamellar keratectomy or penetrating keratoplasty should be considered in those patients who do not respond to medical therapy or those who have recurrent exacerbations on attempted weaning of topical antibiotic therapy. PMID- 9754175 TI - Utility of microdissection and polymerase chain reaction for the detection of immunoglobulin gene rearrangement and translocation in primary intraocular lymphoma. AB - OBJECTIVE: Primary intraocular lymphoma, a non-Hodgkin's lymphoma, is a primary central nervous system lymphoma (PCNSL). Diagnosis is usually made by identifying malignant, large B lymphocytes in the vitreous, eye, brain, and cerebral spinal fluid; however, these cells are few, friable, and difficult to recognize. Recently, clonal heavy chain immunoglobulin (IgH) gene rearrangement and bcl-2 gene translocation have been reported in systemic B-cell lymphoma and are used for the detection of malignant cells and in making a diagnosis. The authors investigated the molecular changes in three eyes and a chorioretinal biopsy specimen of four patients with PCNSL. DESIGN: Human tissue study. MATERIALS: Five ocular specimens of PCNSL were collected. INTERVENTION: The first patient had a diagnostic enucleation of the left eye. The second patient underwent diagnostic chorioretinal biopsy. In the third case, a pair of autopsied eyes with reactive lymphoplasmacytic infiltrates of a patient with acquired immune deficiency syndrome (AIDS) were studied. In the fourth case, an enucleated eye of a patient with AIDS-associated lymphoma was sampled. MAIN OUTCOME MEASURES: The bcl-2 and IgH genes of the lymphoma cells from routine, paraffin-embedded, formaldehyde fixed, or frozen histologic tissue sections were analyzed using microdissection and polymerase chain reaction (PCR) technique. RESULTS: Lymphoma cells obtained from the above four cases showed IgH rearrangement gene in the third framework of the VH region. Bcl-2-associated translocation also was detected in three cases (cases 1, 2, and 4). CONCLUSION: Rearrangement of the IgH gene can serve as a molecular marker for PCNSL. Microdissection allows for procurement and analysis of specific, selected, minute cell populations that are obtained from histologic sections of the complex, heterogeneous tissue. Translocation of IgH and bcl-2, the apoptotic "survival" signal and proto-oncogene, could contribute to the pathogenesis of PCNSL. The combination of microdissection and PCR is a powerful tool for studies of small lesions and cell populations and for understanding disease mechanisms. PMID- 9754176 TI - Long-term survival in choroidal and ciliary body melanoma after enucleation versus plaque radiation therapy. AB - OBJECTIVE: This study aimed to determine whether the long-term melanoma-specific mortality rate of patients with a primary choroidal or ciliary body melanoma treated by enucleation is appreciably lower than that of similar patients treated by plaque radiation therapy. DESIGN: Retrospective, nonrandomized, comparative clinical trial. PARTICIPANTS: A previously reported group of 237 patients, 140 treated by enucleation and 97 treated by cobalt-60 (Co-60) plaque between May 1976 and June 1980, and a residual group of 122 patients, 51 treated by enucleation and 71 treated by Co-60 plaque, were identified by variable-by variable range matching. INTERVENTION: Primary treatment by enucleation or Co-60 plaque radiation therapy was performed. MAIN OUTCOME MEASURES: Melanoma-specific mortality and duration of post-treatment survival were measured. RESULTS: The melanoma-specific mortality rate was substantially worse in the original enucleation subgroup over the entire 15-year follow-up interval; however, differences in baseline prognostic factors between the subgroups are likely to explain the difference in survival curves. After elimination of patients with nonoverlapping values of individual clinical variables to adjust for recognized intergroup differences at baseline, there was no significant or clinically important difference in the 15-year mortality curves of the residual subgroups. The relative rate ratio for the treatment effect in the residual patients was 0.97 (95% confidence interval, 0.51-1.86). There was no late downturn in the survival curve of the plaque-treated patients or late crossing of the curves. CONCLUSION: A large difference in survival between equivalent groups of patients with primary choroidal or ciliary body melanoma treated by enucleation versus plaque radiation therapy appears to be unlikely. PMID- 9754177 TI - Multiplanar imaging in the preoperative assessment of metallic intraocular foreign bodies. Helical computed tomography versus conventional computed tomography. AB - OBJECTIVE: This study aimed to compare the effectiveness of helical computed tomography (CT) versus conventional CT in the preoperative assessment of metallic intraocular foreign bodies on axial, coronal, and multiplanar reconstruction images in clinical routine. DESIGN: Prospective comparative trial, alternate assignment of consecutive patients. PARTICIPANTS: Eighteen patients with penetrating eye injuries and suspected metallic intraocular foreign bodies were studied. INTERVENTION: Alternate patients were assigned to undergo either helical CT or conventional CT in the axial plane. Both the helical and the conventional data were transferred to a workstation, and reconstructions in the coronal and sagittal planes were performed. Additional direct coronal scanning was performed only when necessary for preoperative assessment. MAIN OUTCOME MEASURES: The quality of the directly obtained axial and coronal, as well as the reconstructed coronal and sagittal images, was assessed for each, imaging method based on the ability to detect and accurately localize foreign bodies. The size of the foreign bodies was measured and compared to the actual diameter. Total examination time and radiation dose delivered to the lens were measured for each imaging method. RESULTS: All foreign bodies were detected by each scanning method on the axial, the coronal, and on the reconstructed planes. The quality of the axial images was similar for helical and conventional CT. The helical technique provided high quality reconstructed images comparable in quality to the directly obtained coronal planes in conventional CT. Reconstructions by conventional technique were not useful for preoperative assessment. The examination time for the total orbital volume was 18 seconds for helical CT examinations and 52 seconds for conventional CT examinations. Radiation dose delivered to the lens for the complete examination was 35 mGy for helical CT axial scanning, 56 mGy for conventional CT axial scanning, and 63 mGy for conventional CT coronal scanning. CONCLUSIONS: Helical CT multiplanar imaging offers several significant advantages for the preoperative assessment of metallic intraocular foreign bodies compared to the conventional CT technique in clinical practice, including short examination time, reduced motion artifacts, reduced radiation exposure, and the ability to obtain diagnostically useful coronal and sagittal reconstruction images without the need for additional scanning. PMID- 9754178 TI - Magnetic resonance imaging in pseudotumor cerebri. AB - OBJECTIVE: To determine whether magnetic resonance (MR) imaging can be used to predict the presence of elevated intracranial pressure. DESIGN: Retrospective case series. PARTICIPANTS: Twenty patients with pseudotumor cerebri and 20 control subjects. INTERVENTION: Magnetic resonance imaging. MAIN OUTCOME MEASURES: The presence or absence of the following six neuroimaging signs was measured: (1) flattening of the posterior sclera; (2) enhancement of the prelaminar optic nerve; (3) distension of the perioptic subarachnoid space; (4) intraocular protrusion of the prelaminar optic nerve; (5) vertical tortuosity of the orbital optic nerve; and (6) empty sella. RESULTS: The MR imaging disclosed flattening of the posterior sclera in 80% of patients with pseudotumor cerebri, empty sella in 70%, distension of the perioptic subarachnoid space in 45%, enhancement of the prelaminar optic nerve in 50%, vertical tortuosity of the orbital optic nerve in 40%, and intraocular protrusion of the prelaminar optic nerve in 30%. Each neuroimaging sign was detected in 5% of control subjects, except for enhancement of the prelaminar optic nerve, which was not detected in control subjects. Based on these MR imaging signs, the examiner was able to predict the presence of elevated intracranial pressure in 90% of cases with pseudotumor cerebri and the absence of elevated intracranial pressure in all control subjects. CONCLUSIONS: Elevated intracranial pressure produces a constellation of MR imaging signs that can assist in establishing the diagnosis of pseudotumor cerebri. PMID- 9754179 TI - Repair of exposed hydroxyapatite orbital implant by a tarsoconjunctival pedicle flap. AB - OBJECTIVE: This study aimed to determine a surgical method of management of exposed hydroxyapatite orbital implants. DESIGN: Noncomparative small case series. PARTICIPANTS: Four patients with exposures of hydroxyapatite orbital implants are described. INTERVENTION: A superiorly based tarsoconjunctival pedicle flap is fashioned from the upper eyelid; this flap is inverted and placed over the scleral-covered, exposed hydroxyapatite orbital implant. The flap is divided as a second-stage procedure 4 weeks later. MAIN OUTCOME MEASURE: Mucosal coverage of the previously exposed orbital implant was measured. RESULTS: Four cases are presented with excellent postoperative results at 3 to 18 months' follow-up. CONCLUSION: A suitably fashioned pedicle flap from the upper eyelid is an effective method of managing exposed hydroxyapatite orbital implants. PMID- 9754181 TI - Characteristics of glaucoma drainage implants during dynamic and steady-state flow conditions. AB - OBJECTIVE: This study aimed to ascertain whether the Optimed, Krupin, and Ahmed drainage devices function as valves that vary resistance depending on flow conditions to maintain pressure within a desired range. STUDY DESIGN: Experimental study. INTERVENTION: The three devices and a control cannula were submerged in fluid and perfused with balanced salt solution using a computer driven apparatus that continuously monitors flow (Q) and pressure (P). In one set of experiments, the flow rates were maintained at 2, 5, 10, 25, or 50 microliters/min until steady-state pressures were achieved. In another set of experiments, the flow rate was increased linearly from 0 to 100 microliters/min over 15 to 20 minutes. MAIN OUTCOME MEASURES: The resistance of each implant was calculated from the first set of experiments by dividing the change in pressure (P) by the change in flow (Q) between successive perfusion rates. Flow-pressure curves were plotted from the experiments in which perfusion rate was increased linearly. RESULTS: Resistance remained relatively constant for the cannula (0.18 0.24 mmHg/microliter/min), the Krupin (0.09-0.25 mmHg/microliter/min), and the Optimed implants (0.04-0.08) throughout the tested flow rates. For the Ahmed device, conversely, resistance decreased proportionally (2.86-0.05 mmHg/microliter/min) to the increase in flow. When flow rate was increased linearly from 0 to 100 microliters/min, the Optimed and Krupin devices as well as the cannula generated a linear pressure response with a constant slope. The pressure in the two devices increased at a rate of 0.11 mmHg/microliter compared to 0.23 mmHg/microliter/min for the cannula. The flow-pressure curve for the Ahmed implant was distinct with a steep initial pressure rise and an essentially constant pressure of 12 mmHg thereafter. CONCLUSION: The Optimed and Krupin devices displayed resistance and pressure responses to various flow conditions that were similar to those of a cannula or flow resistor. In these devices, resistance remained relatively stable and pressure increased linearly with flow. The Ahmed device, conversely, functioned as a valve that closely regulated pressure within a desired range by decreasing or increasing resistance as a function of flow. PMID- 9754180 TI - Novel trabecular meshwork inducible glucocorticoid response mutation in an eight generation juvenile-onset primary open-angle glaucoma pedigree. AB - OBJECTIVE: This study aimed to update a large kindred with juvenile-onset primary open-angle glaucoma (POAG) first described in 1940 and to identify the underlying genetic cause of the disease. DESIGN: Molecular genetic study of a single kindred, including clinical examination, retrospective review of clinical and family history records, linkage analysis, and mutation screening. PARTICIPANTS: The retrospective review included 957 members of a single large family. The linkage study included 40 members of 1 branch of the family in which juvenile onset POAG is segregating in an autosomal-dominant pattern. Mutation screening included 15 at-risk family members with juvenile-onset POAG, probands of 40 families with adult-onset POAG, probands of 11 additional unrelated juvenile onset POAG families, and 43 unrelated normal control subjects. INTERVENTION: Clinical and family history records were obtained, ophthalmologic examinations were performed, and blood samples were drawn for use in genotyping. MAIN OUTCOME MEASURES: Allele sizes of microsatellite repeat genetic markers from the vicinity of the GLC1A glaucoma gene on chromosome 1q were assigned based on size fractionation of DNA fragments generated by polymerase chain reaction (PCR). Linkage was established by the method of lod scores. Mutations were identified by determination of the DNA sequence of PCR products amplified from the trabecular meshwork inducible glucocorticoid response (TIGR) gene. Glaucoma status for purposes of linkage and mutation analysis was based on a combination of ophthalmologic examination, clinical records, family history, and previously published information. For some individuals reported in the pedigree, but not included in the genotyping studies, less information was available as presented in the text and tables. RESULTS: Autosomal-dominant POAG was confirmed or reported for 78 members of an 8-generation family. Linkage analysis showed significant evidence for linkage of juvenile-onset POAG in one branch of the family to D1S452 (maximum lod score of 6.42 at a recombination fraction of 0.00) and other markers in the vicinity of the GLC1A gene on chromosome 1q. Screening of the TIGR gene identified a mutation that results in substitution of asparagine for isoleucine at codon 477 near the carboxyterminal end of the protein. CONCLUSIONS: The authors' findings strongly suggest that the juvenile-onset POAG locus in this family is the GLC1A locus and that the underlying cause of the disease is the IIe477Asn TIGR mutation that cosegregates with juvenile-onset POAG in one branch of this large family. Lack of samples from deceased individuals prevented the authors from determining whether reported adult-onset cases in this family could also be attributed to the IIe477Asn TIGR mutation. Absence of the IIe477Asn TIGR mutation from other juvenile- and adult-onset POAG families implies that this TIGR mutation is not a common cause of glaucoma. PMID- 9754182 TI - A randomized, placebo-controlled trial of topical cyclosporin A in steroid dependent atopic keratoconjunctivitis. AB - OBJECTIVE: This study aimed to investigate the therapeutic effect of topical cyclosporin A (CsA) 2% in maize oil as a steroid-sparing agent in steroid dependent atopic keratoconjunctivitis. DESIGN: Prospective, randomized, double masked, placebo-controlled trial. PARTICIPANTS: Twenty-one patients with steroid dependent atopic keratoconjunctivitis were studied. INTERVENTION: Patients used either topical CsA or vehicle four times daily for 3 months in addition to their usual therapy, and the clinical response was used to taper or stop topical steroids when possible. MAIN OUTCOME MEASURES: Steroid drop usage per week, ability to cease steroid use, scores for symptoms and clinical signs, drop side effects, and overall subjective rating of trial drop by patients and clinician were measured. RESULTS: Cyclosporin A had a greater steroid-sparing effect than did placebo. Nine of 12 CsA patients ceased steroids compared to 1 of 9 placebo patients (P = 0.01), the final steroid use was lower in the CsA group (2.6 +/- 1.4 vs. 27.7 +/- 17.7, P = 0.005), and the mean reduction in steroid use was greater for CsA (85.5 +/- 14.7 vs. 13.9 +/- 16.0, P = 0.005). Clinical signs and symptom scores were reduced to a greater level for CsA. Serious side effects were lid skin maceration in one patient using CsA and an allergic reaction in one placebo patient. Marked blurring of vision after drop instillation was common in both groups, but intense stinging was more common in CsA patients (9/12 vs. 1/9, P = 0.01), limiting frequency of drop use. The clinician rated the trial drops as good or excellent more frequently for CsA (11/12 vs. 0/9, P < 0.0001). CONCLUSIONS: Topical CsA is an effective and safe steroid-sparing agent in atopic keratoconjunctivitis and, despite difficulties in patient tolerance, also improves symptoms and signs. PMID- 9754183 TI - Diffuse lamellar keratitis. A new syndrome in lamellar refractive surgery. AB - OBJECTIVE: This study aimed to describe a syndrome that the authors call diffuse lamellar keratitis that follows laser in situ keratomileusis (LASIK) and related lamellar corneal surgery. DESIGN: Noncomparative case series and record review. PARTICIPANTS: Thirteen eyes of 12 patients in whom infiltrates developed in the interface after lamellar refractive surgery were studied. INTERVENTION: Topical antibiotics or corticosteroids or both were administered. MAIN OUTCOME MEASURES: Corneal infiltrate appearance, focality, location, and clinical course were measured. RESULTS: Patients presented between 2 and 6 days after surgery with pain, photophobia, redness, or tearing. Ten cases directly followed either myopic keratomileusis or LASIK. Three cases followed enhancement surgery without the use of a microkeratome. All 13 cases had infiltrates that were diffuse, multifocal, and confined to the flap interface with no posterior or anterior extension. The overlying epithelium was intact in each case. Cultures were negative in the two cases cultured. Ten eyes were treated with antibacterial agents; two eyes had fluorometholone four times daily added to the routine postoperative antibacterial regimen, and one eye had the antibacterial agent discontinued and was treated with topical fluorometholone alone. All infiltrates resolved without sequelae. CONCLUSIONS: A distinct syndrome of unknown cause of noninfectious diffuse infiltrates in the lamellar interface is described. It can be distinguished from infectious infiltrates by clinical presentation and close follow-up. Patients with the syndrome should be spared the more invasive treatment of infectious keratitis. PMID- 9754184 TI - Laser vision correction for low hyperopia. An 18-month assessment of safety and efficacy. AB - OBJECTIVE: This study aimed to assess the efficacy and safety of hyperopic photorefractive keratectomy (PRK) and to evaluate the effect of degree of hyperopia, two epithelial removal methods, and various postoperative patient management techniques on clinical outcomes. DESIGN: Prospective, nonrandomized, open-label clinical trial. PARTICIPANTS: A total of 38 patients with mean follow up of 13.9 months (n = 65 eyes with hyperopia from +1.00 diopter [D] to +4.00 D) participated. INTERVENTION: Hyperopic PRK with the VISX STAR Excimer Laser System was performed. MAIN OUTCOME MEASURES: Spherical equivalent (SE) including vector analysis of SE; uncorrected visual acuity (UCVA); best-spectacle corrected visual acuity (BSCVA); low-, medium- and high-contrast visual acuities; topography; keratometry; pachymetry; and intraocular pressure, haze, and all other potential complications were measured. RESULTS: A total of 80% of eyes were within +/- 0.5 D and all but 1 eye (98%) were within +/- 1.0 D of intended manifest SE at 1 year. There was no induced astigmatism at 1 year. At 12 months, 72% of eyes had UCVA of 20/25 or better and 70% had achieved preoperative BSCVA, with no eye seeing worse than 20/25. These results remained constant at 18 months. There was a tendency toward regression between months 1 and 6 with stabilization of SEs between months 6 and 12. Thereafter, up to 18 months, there was some regression with a mean of +0.31 D, but the number of patients was small. There was one mild decentration and very slight decreases in mean intraocular pressure and central corneal thickness. One patient had grade 1.0 haze develop in both eyes at 12 and 18 months; all other patients experienced trace or no haze. There were no significant complications. CONCLUSIONS: The results of this study support the hypothesis that laser vision correction is safe and effective for treating low hyperopia. The predictability of the hyperopic laser vision correction procedure used in this study was very good. Other than the slower recovery of BSCVA and UCVA seen with this procedure, as compared with myopic PRK, there were no significant complications. The trend toward some later regression needs to be further evaluated in a larger number of patients. Overall, patients were very pleased with the treatment, even in the first 6 months. PMID- 9754185 TI - Mini radial keratotomy reduces ocular integrity. Axial compression in a postmortem porcine eye model. AB - OBJECTIVE: This study aimed to examine ocular rupture force in pig eyes after "minimally invasive radial keratotomy" (MRK) and standard radial keratotomy (SRK). DESIGN: Experimental study. MATERIALS: A total of 71 pairs of pig eyes (51 control eyes) were examined. INTERVENTION: An axial-torsional Materials Testing System (MTS, Eden Prairie, MN) was used to apply blunt force to the corneal surface. A force transducer measured the rupture forces in control eyes and in eyes with MRK or SRK. Five groups of paired eyes were compared: 2.0-mm MRK versus control (N = 12), 3.5-mm MRK versus control (N = 21), 6.5-mm SRK versus control (N = 18), SRK versus 3.5-mm MRK versus 2.0-mm MRK (N = 10). MAIN OUTCOME MEASURE: Ocular rupture force (newtons) was measured. RESULTS: The mean rupture force in newtons was 746.3 for control eyes, 514.2 for 2.0-mm MRK, 353.1 for 3.5-mm MRK, and 246.2 for SRK. Analysis of variance showed a statistically significant difference (P < or = 0.04) between paired comparisons. CONCLUSION: The MRK and SRK significantly weakened ocular integrity compared with control eyes not operated on. MRK required significantly more force to rupture than SRK. MRK eyes, however, ruptured at 50% to 70% of the force required to rupture eyes not operated on. Any patient considering radial keratotomy should be counseled about the risk of greater ocular damage in trauma. PMID- 9754186 TI - Visual function impairments in relation to gender, age, and visual acuity in patients who undergo cataract surgery. AB - PURPOSE: This study aimed to determine the relationship between visual function impairment in 776 patients who had extracapsular cataract extraction with posterior chamber intraocular lens implantation and gender, age, preoperative visual acuity (VA) of both the operative and the contralateral eye, and presence of other ocular disease in the operative eye. DESIGN: Retrospective cross sectional study. PARTICIPANTS: 1139 patients whose medical records were abstracted and who had cataract surgery performed at 1 of 10 participating academic medical centers in 1990. MAIN OUTCOME MEASURE: In the 776 patients who had explicit statements about impairment of visual function documented in their medical records, univariate and multivariable logistic analyses were used to assess the above relationship. RESULTS: The most severe visual functional deficit that justified the cataract operation varied in relation to gender, age, and VA. On bivariate analysis, men were more likely to have impairment with employment, driving, and glare, whereas women were more likely to have impairment with activities of daily living and recreational activities. Significant findings between visual impairment and the independent variables from the logistic regression models included: (1) employment limitation and male gender (odds ratio [OR], 1.92; 95% confidence interval [CI], 1.08-3.40); (2) employment limitation and younger age (OR, 0.12; 95% CI, 0.050-0.28 for ages 70-79); (3) recreational impairment and older age (OR, 2.77; 95% CI, 1.64-4.70 for ages 80+); (4) impairment in performing activities of daily living and female gender (OR, 0.72; 95% CI, 0.53-0.98 for male gender); (5) impairment in performing activities of daily living and worse VA in the operative eye (OR, 5.13; 95% CI, 2.93-9.00 for VA < 20/100); (6) glare-associated impairment and younger age (OR, 0.40; 95% CI, 0.24-0.69 for age 80+); and (7) glare-associated impairment and better VA (OR, 0.16; 95% CI, 0.067-0.38 for VA < 20/100). CONCLUSION: When deciding whether to perform cataract surgery, functional impairment must be considered in relation to the age and the gender of the patient, for the type of functional impairment varies in association with age and gender. PMID- 9754187 TI - Medications and cataract. The Blue Mountains Eye Study. AB - PURPOSE: Corticosteroids are known to cause cataracts, but the effects of other medications on the lens are unclear. The aim of this study was to investigate the relationships between cataracts and a range of medications, including allopurinol, aspirin, chloroquine, diuretics, phenothiazines, and simvastatin. DESIGN: Population-based cross-sectional study. PARTICIPANTS: 3654 individuals 49 to 97 years of age (response rate, 82%) from an urban community near Sydney, Australia, were included. TESTING: Lens photography. PRIMARY OUTCOME MEASURE: Lens photographs were graded for the presence and severity of cortical, nuclear, and posterior subcapsular cataract. RESULTS: After adjusting for numerous potential confounders in ordinal regression models, use of phenothiazines was associated with nuclear cataract (adjusted odds ratio [OR], 2.18; 95% confidence interval [CI], 1.01-4.74); long-term aspirin users (> or = 10 years) had higher prevalence of posterior subcapsular cataract than did nonusers and short-term users (test for trend, P = 0.02); and the antimalarial drug mepacrine was associated with posterior subcapsular cataract (adjusted OR, 3.56; 95% CI, 1.56 8.13). There was a suggestion that use of chloroquine-like drugs for more than 1 year (test for trend, P = 0.12) might also be associated with posterior subcapsular cataract. Antihypertensive medications, cholesterol-lowering drugs, and allopurinol were not associated with any type of cataract. Potassium-sparing diuretics were the only diuretic to show any evidence of an association with cataract (test for trend for posterior subcapsular cataract, P = 0.14). Amiodarone was associated with cortical cataract (age- and gender-adjusted OR, 3.84; 95% CI, 1.01-14.81), but there were too few users to do analyses adjusted for multiple confounders. CONCLUSIONS: Most drugs commonly used in the community do not appear to be associated with cataract. The findings of this study do not support the hypothesis that aspirin protects against cataract. PMID- 9754188 TI - Refractive and visual outcome of hyperopic cataract cases operated on before and after implementation of the Holladay II formula. AB - OBJECTIVE: The primary objective was to evaluate the refractive and visual outcomes in a series of hyperopic cataract cases in which the Holladay II intraocular lens (IOL) power formula was used in conjunction with added eye measurements (measured anterior chamber depth [ACD], lens thickness, and corneal diameter) to improve predictability of refractive outcome. In addition, the impact of use of a double ("piggyback") IOL on refractive outcome was evaluated. DESIGN: Prospective, nonrandomized comparative clinical trial. PARTICIPANTS: A total of 136 consecutive hyperopic primary cataract-IOL cases operated on at in an outpatient eye surgery center were evaluated. The main inclusion criterion was the requirement of at least 30 D of emmetropia power. INTERVENTION: Implantation of a total implanted power calculated using a newly developed (Holladay II) formula, which uses additional eye measurements (measured ACD, lens thickness, corneal diameter) in addition to the axial length and keratometry normally used, was performed. In the first series, IOL powers were chosen using the Lloyd-Gills formula with modifiers; in the second series, powers were chosen using the Holladay II formula option in the Holladay IOL Consultant software. Selection criteria for both series were the same (requiring at least 30 diopters [D] of power for emmetropia). Keratometry and axial length measurements (by immersion) were taken using the same instrumentation and methodology in both series. Predicted postoperative refraction based on the IOL implanted and the method of power calculation used were computed for each case in both groups and compared to the actual achieved refraction. MAIN OUTCOMES MEASUREMENTS: Main clinical outcome parameters evaluated were the postoperative spherical equivalent (compared with the predicted spherical equivalent) and the best-corrected vision. These outcome parameters were evaluated within each surgical series, in the total group of cases (regardless of power calculation method). Further stratification according to the use of single or double implants also was done. RESULTS: In the group using an older formula system, mean preoperative spherical equivalent of 4.79 D was reduced to -0.67 D. Similarly, in the Holladay II group, the preoperative mean of 5.60 D was reduced to -0.58 D. However, there were fewer large deviations between predicted and achieved spherical equivalent in the Holladay II group as indicated by a smaller standard deviation of the absolute deviation (0.47 vs. 0.59), and the range of postoperative refractions was smaller with fewer large overcorrections or undercorrections. However, almost 90% of both groups were within a diopter of the predicted refraction. Visual results were comparable in the two groups. CONCLUSION: Both IOL calculation systems showed good predictability in these extremely short eyes. The Holladay II formula was simpler because it is incorporated into a user-friendly software package (Holladay IOL Consultant) and required only the input of IOL constants and preoperative measurements with no "fudge factor" modifiers. Results within the series using this formula had a tendency toward a smaller standard deviation with fewer outliers. PMID- 9754189 TI - Long-term follow-up of extracapsular cataract extraction and posterior chamber intraocular lens implantation in patients with uveitis. AB - OBJECTIVE: The objective of the study was to determine the long-term outcome of patients with uveitis who underwent extracapsular cataract extraction (ECCE) and posterior chamber intraocular lens (PCIOL) implantation. DESIGN: Retrospective review. PARTICIPANTS: Twenty-eight patients (36 eyes). INTERVENTION: Extracapsular cataract extraction and PCIOL implantation. MAIN OUTCOME MEASURES: Level of best-corrected Snellen visual acuity, change in visual acuity, length of follow-up, long-term findings, and complications. RESULTS: In long-term follow-up (mean, 81.4 months), 94% of eyes had visual acuity improvement compared with preoperative levels. Average change in visual acuity for all eyes was an improvement of 6.4 Snellen lines; 75% of eyes were 20/40 or better. The prevalences of cystoid macular edema (CME), epiretinal membrane (ERM), and posterior capsule opacification (PCO) were 56%, 56%, and 58%, respectively. CONCLUSIONS: Patients with uveitis who are treated with ECCE with PCIOL implantation can have successful visual results in long-term follow-up despite the prevalence of PCO or macular abnormalities such as CME and ERM. PMID- 9754190 TI - Primary intraocular lens implantation for penetrating lens trauma in Africa. AB - OBJECTIVE: This study aimed to audit the surgical strategy of primary posterior chamber intraocular lens implantation for cases of recent penetrating trauma involving the lens in an African population. DESIGN: Retrospective, noncomparative case series. PARTICIPANTS: Seventy-two cases are reported, including all patients who underwent primary intraocular lens implantation for traumatic cataract extraction performed within 1 month of injury between 1988 and 1996. MAIN OUTCOME MEASURES: Demographic characteristics and follow-up attendance rates are analyzed. Surgical technique and the occurrence of intraoperative and postoperative complications are reported. Visual outcomes are reported with detailed analysis for cases of poor visual outcome. RESULTS: Mean age was 14.3 years (standard deviation = 11.1), 57 (79%) were male and 15 (21%) were female (chi-square = 23.66, P < 0.01). Fifty-eight patients (80%) attended for follow-up with a mean follow-up duration of 14.3 months (standard deviation = 17.3). No demographic or surgical differences were identified between attendees and nonattendees. The posterior capsule had been breached by the trauma in 27 (38%) cases, and 15 of these required anterior vitrectomy. Capsular fixation of the implant was achieved in 49% of patients, the remainder having sulcus fixation. Intraoperative rupture of the posterior capsule occurred in four cases. The only common postoperative complication was acute fibrinous anterior uveitis, which occurred in 29 (40%) patients, and 32% of patients followed up for at least 6 months required secondary posterior capsulotomy. This was more common in younger patients (chi-square = 4.2, P < 0.05). Corrected postoperative visual acuities were available for 51 patients, of which 71% achieved 20/60 or better visual acuity. Patients 6 years of age or younger were less likely to achieve 20/60 (chi square = 6.61, P = 0.01). CONCLUSIONS: This surgical strategy has proved successful, producing good visual results and causing no sight-threatening complications. Primary posterior capsulotomy may be appropriate for younger patients. PMID- 9754191 TI - Comparative effects of ketorolac 0.5% or diclofenac 0.1% ophthalmic solutions on inflammation after cataract surgery. AB - OBJECTIVE: Ketorolac tromethamine 0.5% and diclofenac sodium 0.1% ophthalmic solutions are approved for use by the U.S. Food and Drug Administration to avoid excessive postoperative inflammation after cataract surgery and implantation of an intraocular lens. This study compares the efficacy and toxicity of these nonsteroidal anti-inflammatory drugs for the first time. DESIGN: Randomized, double-masked, prospective clinical trial. PARTICIPANTS: A total of 120 patients assigned in equal numbers to 1 of the 2 treatment regimens. INTERVENTION: Treatment with either ketorolac 0.5% or diclofenac 0.1% ophthalmic solutions instilled four times daily for 30 days beginning the first postoperative day after surgery. MAIN OUTCOME MEASURES: Objective (Kowa FC 1000 laser cell and flare meter) and subjective (slit-lamp biomicroscope) measurements of inflammation and toxicity were made and compared at three separate post-operative visits. RESULTS: The anti-inflammatory effects of the two treatment regimens were not statistically different at any of the postoperative visits. Patients tolerated both treatments equally well. CONCLUSIONS: This study shows diclofenac sodium 0.1% and ketorolac tromethamine 0.5% ophthalmic solutions are equally effective and safe for the control of postoperative inflammation after uncomplicated cataract surgery performed by phacoemulsification followed by the implantation of a foldable intraocular lens. PMID- 9754193 TI - Medicines Control Council--statement. PMID- 9754192 TI - A double-masked, placebo-controlled evaluation of 0.5% loteprednol etabonate in the treatment of postoperative inflammation. The Loteprednol Etabonate Postoperative Inflammation Study Group 2. AB - OBJECTIVE: This study aimed to compare the efficacy and safety of loteprednol etabonate (LE) 0.5% to placebo (vehicle) in controlling the anterior chamber cell and flare reaction in patients undergoing cataract surgery with intraocular lens (IOL) implantation. DESIGN: Randomized, double-masked, placebo-controlled, parallel group multicenter study. PARTICIPANTS: A total of 203 patients undergoing elective cataract removal and posterior chamber intraocular lens implantation who, on the day after surgery, exhibited a minimum anterior chamber inflammation score (ACI, sum of cell and flare reaction) rating of 3 (0-9 scale). INTERVENTION: All patients received either LE 0.5% or placebo (vehicle) four times daily in the eye that was operated on for up to 14 days after surgery. MAIN OUTCOME MEASURES: Resolution of ACI by final, on-treatment visit was measured. RESULTS: The proportion of patients with ACI resolved by the final visit was 56 (55%) of 102 in the LE group and 28 (28%) of 100 in the placebo group (P < 0.001). For all the individual components of ACI (cell and flare), as well as other signs and symptoms, the resolution rate and mean change from baseline favored LE. Expanding the efficacy criterion to include patients with mild inflammation at final visit, the efficacy of LE was 95 (93%) of 102 in contrast to 65 (65%) of 100 for placebo. Among the 39 patients who did not complete the study, the majority were discontinued for inadequate anti-inflammatory effect: 25 (25%) of 101 placebo patients and 5 (5%) of 102 of LE patients. The difference in the treatment failure rates, as well the difference in the time course of failures, was both clinically meaningful and statistically significant in favor of LE (P < 0.001). Both treatments were well-tolerated. No clinically significant elevations in intraocular pressure (> or = 10 mmHg) were seen in the LE treatment group. One patient in the placebo treatment group met this criterion. CONCLUSIONS: Loteprednol etabonate showed a clinically meaningful reduction in the signs and symptoms of postoperative anterior chamber inflammation when compared with that of placebo and had an acceptable safety profile compared with placebo. PMID- 9754194 TI - Health science faculties--how committed are they to building a culture of human rights in health? PMID- 9754195 TI - Monitoring drug safety--the power or the yellow form. PMID- 9754196 TI - Lariam--inaccurate information. PMID- 9754197 TI - Why do we need a large study on tobacco-attributed mortality in South Africa? PMID- 9754198 TI - Bringing traditional healers into TB control. PMID- 9754199 TI - More work needed on cannabis. PMID- 9754200 TI - 'Unfettered' hospitalisation in private sector no longer possible. PMID- 9754201 TI - Affordable management of HIV infection in the private sector. PMID- 9754202 TI - Confronting AIDS--a plea for a national dried milk formula. PMID- 9754203 TI - Complications of worm infestation--serious, costly, predictable and preventable. PMID- 9754204 TI - Pernkopf's Atlas--a product of Nazi atrocities perpetrated in Austria during World War II. PMID- 9754205 TI - The desperation of the deregistered doctor. PMID- 9754206 TI - New advances in cystic fibrosis--implications for developing countries. PMID- 9754207 TI - Restoring the honour of our profession. PMID- 9754208 TI - New birth and death registration forms--a foundation for the future, a challenge for health workers? PMID- 9754209 TI - The difficult road to truth and reconciliation--the health sector takes its first steps. Health and Human Rights Project Support Group. PMID- 9754210 TI - Rural health and human rights--summary of a submission to the Truth and Reconciliation Commission Health Sector Hearings, 17 June 1997. PMID- 9754211 TI - Gender audit of health research--10 years of the South African Medical Journal. AB - OBJECTIVE: To examine the extent to which gender bias, which has been identified as a feature of medical research internationally, is present in medical research published in South Africa. DESIGN: A retrospective review was undertaken of 789 articles, 106 letters and 266 editorials in 10 years of the South African Medical Journal (1986-1995). MAIN OUTCOME MEASURES: These were gender of study subjects, proportion of women in the sample, and evidence of analysis of results according to gender of study subject. RESULTS: Forty-eight per cent of articles (377) and 98% of letters (104) did not mention the gender of the sample. Samples that included both genders had significantly fewer women than men, with 80% (297) of such articles and 93% (14) of such letters not presenting a comparative analysis of results. CONCLUSIONS: These findings, similar to those of the international literature, indicate a predominant 'gender blindness' in published works. This precludes investigation of differences in the ways men and women experience disease and differential access to care. PMID- 9754212 TI - Underrecognition and undertreatment of asthma in Cape Town primary school children. AB - BACKGROUND: In view of the high local prevalence of asthma, the extent of recognition and appropriate management of childhood asthma was studied in a large suburban area of Cape Town. DESIGN: Cross-sectional study based on random community sample of schools. METHOD: 1,955 parents of sub B pupils from 16 schools completed a questionnaire, followed by: (i) an interview of the parents of 348 symptomatic children; and (ii) bronchial responsiveness testing on 254 children. The final case group consisted of 242 children with reported asthma or multiple asthma symptoms on both questionnaires. Children in whom asthma was acknowledged were compared with those in whom it was not. RESULTS: Overall, any past or current ('ever') asthma was acknowledged by respondents in only 53% of the children, and current asthma in only 37.1%. While most children had received treatment in the previous 12 months, 66.1% of the recognised group were on current treatment (23.2% on daily treatment), compared with 37% of the unrecognised group (3% daily). Salbutamol and theophylline syrups were the most common types of medication, while inhalers and anti-inflammatory medications were underused. Only a minority of parents reported the child ever having used a peak flow meter, or volunteered knowledge of preventive measures. Current treatment, and to a lesser degree recognition of asthma by parents, were more common among children on medical aid and of higher socio-economic status. CONCLUSIONS: These findings suggest that ways need to be found: (i) to increase the use of current asthma treatment guidelines by practitioners; (ii) to provide access to comprehensive care by children not on medical aid; and (iii) to improve education of parents in home management measures such as severity assessment and avoidance of smoking, allergen and dietary triggers. PMID- 9754213 TI - Lung function in South African children with cystic fibrosis. AB - OBJECTIVE: To determine the pattern of lung function in stable cystic fibrosis (CF) patients and to investigate the relationship of abnormal lung function to demographic variables, CF genotype and pulmonary colonisation with Pseudomonas aeruginosa (PA). DESIGN: A descriptive study done at the CF clinic at Red Cross War Memorial Children's Hospital in Cape Town. METHODS: Data were recorded and pulmonary function testing (PFT) was performed in 42 CF patients. RESULTS: 29 patients (69%) had mild disease, while 11 (26%) and 2 (5%) had moderate and severe disease respectively. Twenty-four patients (57%) demonstrated lower airway obstruction (LAO). Patients with moderate or severe disease were significantly older than those with mild disease (13.3 (3.7) years (mean (SD)) compared with 11.1 (3.0) years (t = 2.1; P = 0.04). PA colonisation status differed significantly with the pattern of lung function (chi 2 = 6.6; P = 0.04) and severity of lung disease (chi 2 = 12.6; P = 0.002). Nine (35%) of the 26 patients tested before and after bronchodilator therapy showed a positive response. CONCLUSION: The majority of patients had mildly impaired or normal lung function, with LAO predominating. A minority of patients were bronchodilator-responsive. PA colonisation may be associated with the development of abnormal lung function and more severe pulmonary disease. PMID- 9754214 TI - Guidelines for the management of chronic obstructive pulmonary disease. Working Group of the South African Pulmonology Society. AB - OBJECTIVE: This guideline has been developed in order to optimise the management of patients with chronic obstructive pulmonary disease (COPD) at all levels of the health care system in South Africa. It contains an action plan for early recognition and appropriate treatment of this common condition. OPTIONS: Treatment regimens are recommended for patients with mild (stage I), moderate (stage II) and severe (stage III) disease. OUTCOMES: Optimal management of patients with COPD may achieve a reduction in breathlessness, improved quality of life, prevention of complications and limitation of disease progression. EVIDENCE: The Working Group comprised mainly pulmonologists, but included an anaesthetist, a pharmacologist and a physiotherapist. Detailed literature review with particular attention to similar guideline documents from Europe and the USA was performed before the meeting. RECOMMENDATIONS: Steps in the management of patients with COPD include early recognition of the disease, smoking cessation, treatment of airflow obstruction with appropriate drugs (singly or in combination), education and pulmonary rehabilitation, and limitation of disease progression and complications. detailed recommendations are made with regard to the use and interpretation of a trial of oral corticosteroid therapy. Indications for hospitalisation, intensive care unit admission and ventilatory support are provided. VALIDATION: This guideline is similar to those recommended by other groups outside South Africa. It was developed by a working group of the South African Pulmonology Society and is endorsed by the Medical Association of South Africa. SPONSORS: The meeting of the Working Group was sponsored by Boehringer Ingelheim. This sponsorship did not influence the activities of the Group. PMID- 9754215 TI - Pancytopenia in a patient with Cryptococcus meningitis. PMID- 9754216 TI - Fishtank water as a source of a rare case of Aeromonas hydrophila septicaemia. PMID- 9754217 TI - [Point of care systems]. PMID- 9754218 TI - [Molecular mechanisms of wound scarring]. AB - A few years ago, the discovery of growth factors, their pharmaceutical obtention at a purified grade, their powerful effects on cells in vitro, resulted in demeasured hopes that they could be applied easily and successfully to the treatment of wounds. Now, the process of healing is still uncompletely understood. The interplay of epithelial and matrix cells, the multiplicity of cell types involved, the huge number of growth factors implicated and the difficulties in describing the specific timing of their action on the cells present in wounds, explain why therapy of wound has not yet been revolutioned. An important distinction must be made between normal healing, which leads to a solid functional, reparation tissue, and scarring which opposes any functional healing by making extensive granulation tissue or even keloids. Recent studies pointed out the necessity of a convenient balance between the concentrations of growth factors present in the wound during the different stages of healing. Excess connective tissue production (scarring) would be more particularly due to an excess of TGF beta 1, whereas predominance of TGF beta 3 conducts to harmonious healing. PMID- 9754219 TI - [Biology of papillomavirus II infections. Their role in the carcinogenesis of the cervix]. AB - The association of human papillomaviruses (HPV), i.e., papillomavirus type 16, with cervical dysplasias and carcinomas is now well established. Additional agents such as sexual behaviour, immunity deficiency, sociodemographic factors, microbiological agents..., are however implicated in the multistage progression from viral infection to cancer. And inactivation of tumor suppressor gene products (p53, p105Rb), oncogene activation (c-myc, c-ras), aneuploidy, karyotypic abnormalities are key events in the tumor progression. Numerous aspects of the biology of human papillomavirus, i.e. natural history, epidemiology, nature and mechanisms of the immune response are under active investigation. Screening strategies of HPV infections (cytology, HPV DNA detection and HPV antibody detection) demonstrated their efficacy in many countries, while prophylaxy and treatment of these infections by vaccines are still under development. PMID- 9754220 TI - [Etiology and diagnostic of viral bronchopneumonias]. AB - Community viral bronchopneumonias are frequent, mainly in children, and can be associated to all respiratory viruses: influenza- and parainfluenzavirus, respiratory syncytial virus, adenovirus, rhinovirus. The diagnostic method which proves viral infection of the respiratory tissues is selected as the direct detection by an immunofluorescence assay of viral infected cells in respiratory samples. In them, viral isolation or nucleic acid detection by PCR provide an amplification of the viruses. By using PCR-hybridation techniques viral detection is overall increased of 1.5 times for respiratory syncytial virus, 1.9 for parainfluenzavirus 3, 4 for rhinovirus and 10 times for adenovirus. This increased sensitivity raises questions about the meaning of the detection of viral sequences in nasal aspirates, with or without clinical signs. Cytomegalovirus (CMV) is a major agent of pneumonia in immunocompromised patients. All virological markers of CMV infection have to be sought (antigenemia, viremia...), but specific inclusions in pulmonary cells are the single diagnosis criteria. As pulmonary biopsies are rarely available and CMV inclusions rarely found in BAL, it has been reported useful to look for high viral loads or late m-RMA transcripts in these samples. Adenovirus pneumonia are unfrequent in these patients and mostly associated to rare or atypical strains. Such PCR-hybridization systems deserves also to be used in these cases. PMID- 9754221 TI - [Thyroglobulin assay ambiguities]. AB - Thyroglobulin immunometric "sandwich" assays (IMA) have taken over competitive radioimmunoassays, but this assay remains problematic. A human thyroglobulin reference material (CRM 457) has been prepared but is not widely used. That constitutes the main cause of very marked between kit variability of thyroglobulin results. High-dose hook effect, which can falsely decrease the result of a serum with high concentration of thyroglobulin, is not exceptional in one-step assays and should be systematically checked. Selection of monoclonal antibodies with no cross-reactivity with anti-thyroglobulin autoantibodies or of polyclonal antibodies with very high affinity, have reduced the frequency of interference due to autoantibodies, but did not abolish it. Recovery test is used to detect such interference, but with insufficient sensitivity. In fact, recovery determination can be influenced by the nature of thyroglobulin added to the serum (not identical to endogenous thyroglobulin), by the delay of incubation of exogenous thyroglobulin and serum autoantibodies and by the amount of added thyroglobulin. In addition, recovery is often wrongly expressed as the observed/theoretic ratio of final concentrations instead of added concentrations. In untreated Graves' disease patients and despite normal recovery test, thyroglobulin measured by IMA is lower when anti-thyroglobulin autoantibodies are present. Consequently, thyroglobulin result should be interpreted in function of presence or absence of autoantibodies. Development of total (free and autoantibody bound) thyroglobulin assay would be useful to evaluate assay and recovery test performances. PMID- 9754222 TI - [Evaluation of hemostasis in venous thromboembolism pathology]. AB - Thromboembolic disease results from an hypercoagulable state and multifactorial causes may lead to hypercoagulability. Thrombogenic risk factors can be acquired and/or inherited. For each thrombophilic patient, the main clinical features retained are: the patient age, the familial history, the recurrence of thromboembolic events, an unusual site of thrombosis. Anti-phospholipid antibodies, which are considered as acquired thrombogenic risk factors, can be detected with coagulation tests and/or Elisa methods. The association of antiphospholipid antibodies with thrombosis is defined as the anti-phospholipid syndrome. Last decades, genetic risk factors were identified. First of all, antithrombin, protein C and protein S deficiencies were described. These deficiencies are involved in about 10% of patients who develop thrombosis before the age of 50. In 1993, a new genetic risk factor was discovered: activated protein C resistance which is due to the Q506 mutation in factor V. This defect represents the most prevalent abnormality of inherited thrombophilia, affecting 20 to 40% of thrombophilic patients. Interestingly, hyperhomocysteinemia, known as potentially predisposing to arterial disease, was also recognized as a risk factor for venous occlusive disease. Several genes encoding homocystein metabolism enzymes, such as cystathionine beta-synthase or methylenetetrahydrofolate reductase are concerned. Establishment of a causal association between the presence of a biological abnormality and the occurrence of thrombosis may lead to an adapted prophylaxis whatever the risk situation. PMID- 9754223 TI - [Bcr-abl translocation: diagnostic methods and clinical value]. AB - The t (9;22) translocation is present in about 95 per cent of chronic myelogenous leukemia and in a significant subset of acute leukemias, mainly of the lymphoid subtype. This chromosomal rearrangement leads to the fusion of the bcr and c-abl genes and to the transcription of leukemia-specific bcr-abl mRNAs. The accurate identification of the t (9;22) translocation relies on cytogenetics (conventional or Fish) and molecular techniques. The detection of residual Ph positive cells following bone marrow transplant or interferon therapy is critical and has relevant therapeutic implications. PMID- 9754224 TI - [Serum benzodiazepine research by the immunoenzymatic method in emergency toxicology: test interpretation aids]. AB - The analyst in toxicology is daily confronted with blood benzodiazepine research. The sensitivity of the immunoenzyme technique initially designed for urine testing has been improved. Nevertheless some discrepancies persist between clinical and biological observation. This work based on kinetic characteristics of benzodiazepine and analytical sensitivity indicates drug quantities which should be ingested to have a positive test. These quantities are computed for a man and a woman of average weight of 70 kg and 50 kg and for a child weighing 13 kg (about three years old). They are then compared to quantities usually considered as toxic. Nevertheless, these values have to be adapted on a case to case basis. As a general observation, analytical sensitivities for benzodiazepine are variable but the immunoenzyme technique ensures a good covering of real toxic quantities of the drug. Flunitrazepam is the one which has the worst covering probability. PMID- 9754225 TI - [Apolipoprotein (a) isoform size determination. Value and limits of high resolution phenotyping by agarose gel electrophoresis]. AB - High-resolution methods using agarose gel electrophoresis followed by immunoblotting have been recently developed for the measurement of the apolipoprotein (a) size isoforms. Despite the high sensitivity of these methods, a variable proportion of isoforms remain undetected. Four primary antibodies were compared for their ability to detect a large number of isoforms, and were found to express equal reactivity irrespective of the apo (a) size. Comparison of the data obtained both by phenotyping and by genotyping led to the identification of 15 artefactual bands. All these nonspecific bands were of low intensity and occurred when more than 50 ng of Lp (a) were loaded on the gel. The visualization of the isoforms was performed by labelling apo (a) with a peroxidase conjugated antibody coupled to a luminescent substrate, thereby allowing quantification of the relative expression of each isoform by densitometry. There is currently no standardized nomenclature for the apo (a) isoforms: a procedure is proposed using a commercially available standard to express the results in kringle number. PMID- 9754226 TI - [Evaluation of a dot-blot technique for the detection of Epstein-Barr virus antibodies (IgG anti-viral capsid antigen)]. AB - Infectious mononucleosis syndrome is caused by several infectious agents, including Epstein-Barr virus (EBV), cytomegalovirus (CMV), and Toxoplasma gondii. The ImmunoDot EBV VCA-IgG test (Biomedical Diagnostics) is a dot-blot enzyme immunoassay, which determines the presence of heterophile antibodies and IgG antibodies directed against Epstein-Barr viral capsid antigen (anti-VCA), CMV and T. gondii. This test is simple, unitary, ready-to-use, and rapid (40 minutes), requiring approximately 10 microliters of serum or plasma or 20 microliters of whole blood. The aim of this study was to compare the results of this technique for the determination of anti-VCA IgG antibodies with those obtained by the reference technique using indirect immunofluorescence on 185 serum specimens. A sensitivity of 99.2%, a specificity of 92.4%, a correlation with indirect immunofluorescence of 97.3%, a the absence of cross-reactions with CMV, T. gondii, and herpes simplex viruses and the absence of interference with antinuclear antibodies were the main results of this comparative study. In association with another test (Monolert 2TM) detecting IgM and IgG antibodies against Epstein-Barr nuclear antigen, the dot-blot method represents a useful screening strategy for EBV response. PMID- 9754227 TI - [Comparison of five phenotyping methods for the detection of methicillin resistance in Staphylococcus aureus]. PMID- 9754228 TI - [Control of nosocomial infections: value of a combined analysis of bacterial ecology and computerized medical department information]. PMID- 9754229 TI - [Cryptates applied in tumor marker assays (CEA, AFP, CA 15.3, CA19.9): evaluation of Kryptor (Cis-Bio) analyser]. PMID- 9754230 TI - [Postmortem production and utilization of ethanol in two body liquids]. PMID- 9754231 TI - [Hypereosinophilia and hematologic analysers "formula approach"]. PMID- 9754232 TI - [Chronic lymphoproliferative disorder with villous cells: tricholeucocytosis or splenic lymphoma with villous lymphocytes?]. PMID- 9754233 TI - [Evaluation of a new assay for the direct determination of HDL-cholesterol]. PMID- 9754234 TI - [Venous thrombosis in a pregnant woman heterozygous for factor V Q506 mutation]. PMID- 9754236 TI - [Apolipoprotein E and Alzheimer's disease]. PMID- 9754235 TI - [Recommendations concerning point-of-care systems. "Point-of-care systems" work group of the French Society of Clinical Biology and National College of Hospital Biochemistry]. PMID- 9754238 TI - [Diagnosis of Alzheimer's disease]. AB - Alzheimer's disease is a neurodegenerative disorder leading to cognitive impairment (amnesia, aphasia, apraxia and agnosia). The prevalence of this age related disabling illness is increasing. During the course of the disease, the clinical diagnosis will be that of "possible Alzheimer's disease", then "probable Alzheimer's disease". But the diagnosis of "definite Alzheimer's disease" requires a post-mortem brain examination and the demonstration of numerous senile plaques and neurofibrillary tangles in hippocampal and association cortical areas. The neuropathological examination confirms probable Alzheimer's disease clinical diagnosis in 85% of the cases examined in medical schools. However, with much more than 15% errors, the early diagnosis of Alzheimer's disease must be improved since it is a key factor for the therapeutic approach, and more especially for the efficiency of drug trials. At the present time, there are new leads for a biological diagnosis in the blood or the CSF. However, the natural (and molecular) history of Alzheimer's disease points out that all biochemical dysfunctions remain in the CNS, and more particularly in association brain areas. This is demonstrated using reliable biochemical markers such as A beta and pathological Tau proteins, which are the basic components of amyloid deposits and neurofibrillary tangles, respectively. Also, a genetic diagnosis can be performed in half of familial autosomic dominant Alzheimer's disease, which represents less than 1% of all Alzheimer's disease cases. Together, this shows that there is a lack of reliable Alzheimer's disease markers. The search for new specific markers (clinical, epidemiological, genetic, biochemical, biological) must go on. PMID- 9754239 TI - [Therapeutic use of hematopoietic growth factors. I. Erythropoietin and thrombopoietin]. AB - Modern molecular haematology is characterized by the great strides made in the use of cytokines, especially haematopoietic growth factors. These factors constitute a heterogeneous group of molecules that ensure the survival, proliferation and differentiation of the haematopoietic cells. Present detailed knowledge of the structure of the main haematopoietic growth factors and their receptors, and of the cloning and sequencing of their genes, permits the use of genetic engineering to produce recombinant human growth factors whose therapeutic applications have raised very great hopes for clinical haematology. Data obtained from several clinical studies have allowed the use of some of these molecules in France. This is the case concerning erythropoietin (Eprex, Recormon), G-CSF (Neupogen, Granocyte) and GM-CSF (Leucomax), each with specific uses. Others haematopoietic growth factors, such as stem cell factor (SCF) are presently evaluated for their clinical interest. Finally, interleukin 3 (IL3), whose in vitro activities seemed to be of potential interest, has been evaluated during clinical studies. Its toxicity and lack of specificity have been evidenced and do not allow its present utilization. The first part of this review is focused on the general structure and biological activity of haematopoietic growth factors and presents the actual therapeutic field of the use of erythropoietin and the promising application of recombinant thrombopoietin. PMID- 9754240 TI - [Hemostasis and human immunodeficiency virus (HIV) infection]. AB - Various coagulation abnormalities were reported in HIV-infected patients. Cases of severe thrombocytopenia were first observed in contaminated homosexual males, as well as prolonged APTT due to the presence of lupus-like anticoagulant with a frequency in the range 8 to 70% of the studied patients. Even if lupus anticoagulant could be evidenced in asymptomatic patients, it frequently occurred during acute opportunistic infections such as Pneumocystis carinii. More recently, increased prevalence of protein S and heparin cofactor II deficiency, two physiological coagulation inhibitors were demonstrated in HIV-infected patients. The same applied for hypoalbuminemia-related fibrin polymerization defects which induced prolonged thrombin and reptilase clotting times. Abnormalities of the fibrinolytic system were also reported, such as increased levels of both tissue-type plasminogen activator and type 1 plasminogen activator inhibitor or decreased levels of histidine-rich glycoprotein. Even if the acute phase response could play a key-role, the pathogenesis of these abnormalities is not fully understood, so far. In addition, their clinical consequences have not been extensively investigated, but hemorrhage appeared to be uncommon. Moreover, D-dimer levels were found increased in HIV-infected patients, suggesting that HIV infection might be associated with a so-called prethrombotic state, which could lead to clinical thrombosis in some HIV-infected patients (2%). PMID- 9754241 TI - [Laboratory follow-up in menopause]. AB - Menopause is not an illness. Nevertheless, nowadays, it is medically approached. We know now with precision the consequences of the estrogen deficiency on about fifty woman's wellbeing, on her metabolism, her cardiovascular system, her bones. A substitutive hormonal therapy is more and more often proposed in order to correct the immediate functional disorders and to prevent the long term consequences (cardiovascular diseases and osteoporosis especially). What is the place of biology in the follow-up of a menopausal patient? Even if clinic observation is predominant for the diagnosis of menopause, plasmatic FSH and estradiol assays are of precious aid in particular cases (hysterectomised patients for example). Whether or not we consider a substitutive hormonotherapy, menopause installation is a good opportunity for detection of metabolic diseases, beginning often in this part of life or clinically suspected. Before beginning a substitutive treatment, biological assays contribute to define the patients's metabolic profile in order to adjust the choice between oral or percutaneous estrogenotherapy and to detect contra-indications. The biological markers of osteoporosis risk are objective arguments to incite indecisive women for beginning or for continuation of a treatment. Under hormonotherapy, plasmatic estradiol's assay should aid to confirm the correct adequation of substitutive doses, but in practice it is not much used: clinical examination is generally adequate to detect under or over dosage. In post-menopause, wether or not she receive a substitutive hormonal therapy, every women should have the benefit of a regular biological follow-up, in the same way as a mammography and cervical smears are recommended. Age being by oneself the main factor for metabolic risk, early detection and early correction of abnormalities will be the main part to ensure quality, for women and also for men, of the second half time of their life. PMID- 9754242 TI - [In vitro and in vivo inhibition of HIV1 replication by retroviral transfer of interferon alpha, beta, or gamma genes: application to gene therapy of AIDS]. AB - Somatic gene therapy is defined as the transfer of a heterologous gene into an organism for the purpose of correcting a genetic defect or providing a new therapeutic function to the target cell and thus inducing a cure or improving associated symptoms. While encouraging results have been generated by recent clinical evaluation of combination of anti-viral drugs, Aids still constitute an obvious candidate among the infectious diseases which might be treated by gene therapy. We have therefore chosen to develop and evaluate a gene therapy strategy based on the transfer into human target cells of HIV1-inducible interferon (IFN) alpha, beta or gamma genes. In a preliminary study, myeloid U937 cell lines transfected with expression vectors containing the IFN alpha, beta or gamma genes under the control of the long terminal repeat (LTR) sequences of HIV1 were shown to be strongly resistant against an in vitro and in vivo (in HIV1 challenged SCID mice model) HIV1 infection. This cellular resistance was correlated with a strong induction of transgenic IFN synthesis and for IFN gamma, with a defect of HIV particles maturation. Secondly, construction and production of high titer retroviral vectors containing Tat-inducible IFN genes allowed efficient transduction of lymphoid cell lines and human primary lymphocytes. These transduced cells were shown to be highly resistant against laboratory and primary HIV isolates. Taken together, our in vitro and in vivo results suggest that HIV1 inducible IFN gene therapy can be beneficial to HIV-infected individuals provided the fact that methods are developed that allow the efficient transduction of human hematopoietic stem cells. PMID- 9754243 TI - [Decarboxyprothrombin: importance in the diagnosis of hepatocellular carcinoma]. AB - We have studied the value of decarboxyprothrombin assay, in association with that of alpha-foeto-protein (AFP), for the biological diagnosis of hepatocellular carcinoma. Levels of decarboxyprothrombin and AFP were measured in 60 patients divided into two groups: 37 patients with hepatocellular carcinoma from liver cirrhosis, confirmed by histology; 23 patients with liver cirrhosis, but having not developed hepatocellular carcinoma. The cirrhosis was in most of cases consecutive to hepatitis B or hepatitis C infection, or of alcoholic origin. Levels of decarboxyprothrombin were also determined in a control group of 50 healthy subjects. Plasma decarboxyprothrombin concentrations were measured by enzyme immunoassay. All normal subjects had levels of decarboxyprothrombin below 2 micrograms/l. Out of 37 patients with hepatocellular carcinoma, 24 (64.9%) showed elevated decarboxyprothrombin levels, while this marker was increased only in 26% of cirrhotic patients. Decarboxyprothrombin and AFP levels are elevated in 48.6% of hepatocellular carcinoma, normal in 16.2% of hepatocellular carcinoma and dissociated in 35.2% of cases; respectively 18.9% and 16.2% of patients with hepatocellular carcinoma have either high AFP level or high decarboxyprothrombin level. The simultaneous determination of decarboxyprothrombin and AFP appear to be useful, since the combination of the two markers allows the detection of 83.8% of hepatocellular carcinoma, while the detection rate is only 67.5% with using AFP alone. No significant correlation was observed between plasma decarboxyprothrombin and serum AFP levels. PMID- 9754244 TI - [Doxorubicin and cisplatin genotoxicity: search for a real indication using the micronucleus test]. AB - Doxorubicin and cisplatin are two major anticancer drugs, and are also known to be mutagen. Using short term mutagenesis tests, the cytokinesis-block micronucleus test and chromosome aberrations test, a study of the cytotoxicity and the mutagenicity of these two drugs has been aimed to determine a genotoxic of reference for these tests. Cisplatin and doxorubicin were genotoxic and gave positive results with the two tests. Since cisplatin was more cytotoxic than doxorubicin for a same genotoxicity, doxorubicin has been selected as a positive control for these two short-term mutagenesis tests. A study of the individual variability in the response to in vitro doxorubicin exposure was made using the cytokinesis-block micronucleus test, applied to cultured T lymphocytes form healthy subjects and cancer patients. Micronucleated cell rate before (T0) and after in vitro exposure to doxorubicin (T1) were determined in the two groups of subjects. Micronucleated cell rates T1 were significantly higher than T0 for healthy subjects and cancer patients. A calculated sensitivity index [(T1)-(T0)] is proposed to evaluate the individual sensitivity to the positive control doxorubicin. PMID- 9754245 TI - [Criteria for addition of apolipoprotein B measurement during routine health screening examinations: validation in a Stanislas cohort. The Biologist's Group of the Health Examination Center]. AB - The purpose of this study was to determine the criteria in which apolipoproteins AI and B should be performed during periodic health screening examinations. Clinically, the results of apolipoproteins AI and B are most useful when there are minor lipid perturbations (cholesterol and triglycerides), but their routine determination is not justified. Several mathematical models, defined by discriminate factorial analysis, have been studied. The one based uniquely on cholesterol and triglyceride concentrations was the most efficient in identifying patients in whom apolipoprotein B should be determined. In contrast, no model was found for the indication of apolipoprotein AI determination. Data from a population different from that used for the model establishment were used to validate the model. This strategy, determining criteria for additional measurement of apolipoprotein B in a general population, is of clinical interest because it may provide complementary information for subjects with atherogenic risk. Moreover, it is of economical interest because it has the potential of limiting complementary testing and its associated costs. PMID- 9754246 TI - [The role of cardiac troponin I, total CK-MB and myoglobin among traditional cardiac markers and their economic aspects]. PMID- 9754247 TI - [Preserving blood specimens before analysis of the latest biochemical parameters]. PMID- 9754248 TI - [Detection of eosinophil granulocyte myeloperoxidase deficiency]. PMID- 9754249 TI - [Measurement of complement C4 fraction in Array 360: beware of monoclonal IgM!]. PMID- 9754250 TI - [Varicella-zoster virus meningo-encephalomyelitis without skin eruption]. PMID- 9754251 TI - [An autochthonous infection by Vibrio cholerae non-01 and non-0139]. PMID- 9754252 TI - [Sweat test]. PMID- 9754253 TI - [Why is there an analytical culture in a medical laboratory?]. PMID- 9754254 TI - [Lack of biological expertise in certain medical publications]. PMID- 9754255 TI - [Oxygen free radicals]. PMID- 9754256 TI - [Glycated hemoglobins: the time for standardization has arrived]. PMID- 9754257 TI - [A registry for European biologists: why create it?]. PMID- 9754258 TI - [Therapeutic use of hematopoietic growth factors. II. GM-CSF and G-CSF]. AB - The second part of this review on haematopoietic growth factors is focused on the therapeutic use of GM-CSF and G-CSF. Such therapeutic applications have raised very great hopes for clinical haematology. However, it should not be forgotten that these haematopoietic growth factors, which are very costly, are powerful two edged weapons capable of triggering a cascade of reactions, and have a field of activity that often goes beyond the single highly specific property which it is hoped they possess. The risks and costs of their use are currently being evaluated. Waited developments concerning these molecules focus on three axes: a best use of factors already commercialized, especially concerning adaptation of posologies and new indications, the development of hybrid molecules from already known haematopoietic growth factors, possessing the advantages of respective factors, but not their disadvantages, the discovery of new haematopoietic growth factors with potential therapeutic application. PMID- 9754259 TI - [The biology of papillomavirus infections. III. Immune response]. AB - Infection with the human papillomaviruses, especially with oncogenic HPVs increases the risk for development of precancerous and cancerous lesions of the cervix. The immune response of the host is likely to be an important factor in determining regression or progression of papillomaviruses-associated lesions. Systemic IgG and IgA response is classically associated with current or past papillomavirus infections. A deficiency in local cellular immune response is however frequently observed and linked to a decrease of cytokine synthesis by infected cells, a reduction or loss of MCH I molecules and a defect in antigen presentation to cytotoxic T lymphocytes. Although secretory immunoglobulins are generated locally in response to papillomavirus infections, humoral immunity in the female genital reproductive tract seems to be inefficient. The papillomavirus infections would lead to a decrease in cellular immunity which could be favourable to viral latency and/or precancerous and cancerous lesion development. PMID- 9754260 TI - [Strategies for identification of secretases implicated in Alzheimer's disease]. AB - In Alzheimer's disease, cortical areas of affected patients are invaded by extracellular proteinous deposits called senile plaques, the main component of which is called amyloid beta-peptide or A beta. This peptide derives from the proteolytic attack of a precursor, the beta-amyloid precursor protein, by two enzymes called beta- and gamma-secretases. Alternatively, beta APP can be cleaved by an additional activity named alpha-secretase that occurs inside the A beta sequence, thereby precluding its formation, and concomitantly liberating a secreted fragment, namely APP alpha. Therefore, secretases seem to play a key role in the control of physiological and potentially pathogenic beta APP catabolites and could be envisioned as possible therapeutic targets in Alzheimer's disease. Here, we describe possible experimental approaches to identify such proteolytic activities. PMID- 9754261 TI - [Bacteriological and clinical aspects of corynebacterium]. AB - The microbiologists use the term corynebacteria to describe aerobically growing, asporogenous, irregularly sharped gram-positive rods. They comprise strictly aerobic bacteria isolated from environment as well as preferentially anaerobic bacteria found in clinical specimens. A large part of these bacteria is considered as commensal of skin and mucous membranes. This group of organisms has recently been subjected to considerable taxonomic revisions, which have resulted in the proposal of several new species, many of them representing previous Centers for Diseases Control coryneform groups. Moreover, recent investigations demonstrated the existence of a pathogenic role for some of them. These bacteria comprise well-known pathogens such as C. diphtheriae responsible for diphtheria, Actinomyces spp. responsible for actinomycosis and Arcanobacterium haemolyticum recovered from pharyngitis, but other corynebacteria were related to particular infections. For example, the lipophilic and antibiotics multiresistant species Corynebacterium urealyticum and C. jeikeium were found to be responsible for urinary tract infections and septicemias, respectively. The recently described species Turicella otitidis was found to be implicated in otitis media and C. seminale were recovered from genital specimens of male patients. Implantation of material devices, use of broad-spectrum antibiotics led to an increase of sepsis due to the species C. jeikeium and C. amycolatum. Many of the new Actinomyces species grow well under aerobic conditions and are often implicated in various abscesses. Moreover an increase of immunocompromised patients led to the development of infections due to the aerobic actinomycete Rhodococcus equi. The association of some corynebacteria with particular diseases should prompt the microbiologist to identify these bacteria when they are encountered in a pathogenic situation. Identification of the major part of corynebacteria isolated from clinical specimens can now be achieved by using recent schemes. PMID- 9754262 TI - [Cytokines and allergic response]. AB - Allergic reactions are under the control of several events that occur sequentially following allergen exposure, recognition by the immune system, IgE production and their interaction with effector cells bearing Fc epsilon receptors. The lymphocyte activation in response to allergens determines the intensity and the nature of the immune response. Cytokines produced by T (and non T) cells are involved in the polarized development of the specific immune response. In particular, type 1 and type 2 cytokines are responsible for the control of the different steps during allergic reactions. Th2 cytokines and particularly IL4 are responsible for switching the immunoglobulin synthesis by B cells to IgE production. They also play a key role in the activation of effector cells that occurs following allergen interaction with fixed specific IgE and participate to the local inflammatory reaction. Cytokine profile determination appears to represent a helpful laboratory parameter in the understanding of the mechanisms underlying allergic diseases. The development of new technological tools may allow the use of cell activation parameters, and cytokine profiles determination in clinical biology. This review aims to analyze the involvement of the cytokine network in the mechanisms leading to IgE production and the involvement of cytokines in effector mechanisms of allergic reactions. It also analyses the potential use of cytokine profile determination for diagnosis purpose and survey of immune desensitization of allergic diseases. PMID- 9754263 TI - [Evaluation of lipid peroxidation by measuring thiobarbituric acid reactive substances]. AB - Malondialdehyde assay is the most generally used test in the appreciation of the role of oxidative stress in disease. Malondialdehyde is one of several products formed during the radical induced decomposition of polyunsaturated fatty acids. Most often, malondialdehyde assay used its reactivity at high temperature and low pH, towards thiobarbituric acid. This reaction is very sensitive but its specificity, even with improvement of pre-analytical (sampling, preservatives), and analytical stages (fluorescence, HPLC) is still a matter of debate. At present, the concept of "thiobarbituric acid reactive substances" (TBARS) have merged and progressively replaced the initial malondialdehyde assay. In this review, we presented the main results concerning the assays of TBARS and malondialdehyde in blood and different biological medium. In the future, oxidative stress appreciation will need the precise analytical determination of different molecules triggered by free radicals. The TBARS assay should be considered as a global test, allowing a global approach of lipoperoxidation whereas specific determination of malondialdehyde can only appreciate one of the end-product formed during oxidative stress. PMID- 9754264 TI - [Blood and leukocyte glutathione and glutathione S-transferase: relationship to cholesterolemia in healthy volunteers]. AB - Hypercholesterolemia increases the oxidation of low density lipoprotein (LDL) which subsequently leads to atherogenesis. The oxidized LDL are also known to increase in vitro macrophage synthesis of glutathione. The purpose of this study was to investigate the relationship between lipid parameters and the glutathione system (glutathione, glutathione S-transferase) in total blood and within leukocytes. The glutathione and glutathione S-transferase were evaluated by spectrophotometric methods in sixty-two healthy volunteers (32 women, 30 men, mean age 39.9 +/- 7.7). No correlation was found between the level of blood cholesterol and the values of the blood glutathione system. However, a positive correlation between the values of glutathione and glutathione S-transferase in leukocytes and the blood cholesterol level was only found in women (r = 0.55 and r = 0.50 respectively, p < 0.01). We also found in men a positive correlation between body mass index and glutathione S-transferase in total blood and within leukocytes (r = 0.38, p < 0.05, r = 0.5, p < 0.01 respectively). No correlation was found between age, smoking and the values of the glutathione system. Our results suggest that the glutathione system in leukocytes is related to blood cholesterol levels. The fact that this positive correlation was only observed in women points to a possible role of estrogens in the regulation of the glutathione system which merits to be further studied. PMID- 9754265 TI - [Characterization of specific IgG, IgM, IgA and IgE isotypes in profound candidiasis]. AB - Enzyme-linked immunofiltration assay technique (Elisa) has been applied to the characterization of G, M, A and E anti-Candida antibodies isotypes specific to cell wall mannans in 201 sera from 126 patients. These sera were studied at the same time using Co-immunoelectrodiffusion and indirect immunofluorescence. In 18 of 21 patients with systemic candidiasis, Elisa demonstrated the presence of antimannan IgG antibodies in sera contemporary of Candida positive blood culture. These IgG were associated with antimannan IgM, A and E in 15 patients. In 37 patients colonized with Candida, used as negative controls, antimannan IgG were detected in 3 cases, and in 2 were associated with specific IgMs. The sensitivity and specificity of Elisa IgM and IgA in the diagnosis of systemic Candidiasis were 85.7% and 81%, respectively. The kinetic study shows that the different isotypes appeared most of the time simultaneously. The evolution of the 4 isotypes beyond the acute episode was variable and without correlation with the clinical status. The decrease of IgG was slower than the one of IgM, IgA or IgE. The systematic research, in at risk patients, of antimannan antibodies using Elisa required simple technology. A simple method should allow to aim at other functional antigens which could be used in a quantitative manner to determine the efficacy of the medical treatment. PMID- 9754266 TI - [An atypical case of antiphospholipid antibody syndrome]. PMID- 9754267 TI - [Role of the laboratory scientist in the diagnosis and treatment of acute lymphoblastic leukemia in children]. PMID- 9754268 TI - [Identification of major yeasts of clinical importance: use of Chromager media for Candida]. PMID- 9754269 TI - [Sideroblastic refractory anemia type myelodysplastic syndrome in a 91-year-old man]. PMID- 9754270 TI - [A new immunosorbent syphilis serodiagnostic technic: an ideal candidate for the replacement of the Nelson and Mayer test]. PMID- 9754271 TI - [Daily practice in clinical enzymology: the danger of utilizing conversion factors tied to measured temperature]. PMID- 9754272 TI - [Microbiological diagnosis in dermatology. Dermatological days of Paris, 3-6 December 1997]. PMID- 9754273 TI - [How to detect, diagnose and survey hepatitis C? Biological tests used by the physician in current practice]. PMID- 9754274 TI - [Medical analysis laboratory accreditation. Office of the French Society of Clinical Biology]. PMID- 9754275 TI - [Apoptotic cell death in response to dengue virus infection: what are the consequences of viral pathogenesis?]. AB - Dengue is a human disease of viral etiology which may be fatal in its hemorragic form. It is widely spread in the tropical areas of the different continents and has been dramatically expanding over the past 30 years. Although an immunological disorder is thought to be involved in dengue physiological symptoms, the pathogenesis of dengue hemorragic fever has not yet been elucidated. Whether the immune response is deleterious or beneficial to the host remains a matter of debate. Other factors, related to virus replication in specific host cells, could also contribute to the severity of the disease. Apoptotic cell death is one of the important consequences of dengue virus infection both in vitro and in vivo. Dengue replication triggers apoptotic signals in neurons and hepatocytes although the original effectors and kinetics differ. Implications of the ongoing apoptotic processes in viral pathogenesis will be further discussed. PMID- 9754276 TI - [In vivo evaluation of insulin sensitivity and clinical applications]. AB - During the past decade, the potential implications of insulin resistance were recognised by clinicians ranging from endocrinologists to cardiologists. Central to this expanding interest is Reaven's hypothesis that tissue resistance to the effects of insulin is a factor linking various metabolic disorders and coronary heart disease. This review critically describes the different approaches for the evaluation of insulin sensitivity in vivo. Qualities and limitations of several investigative techniques are discussed, such as anthropometric indexes, basal biological indexes, insulin suppression tests and insulin tolerance tests. The two most widely used methods for quantifying insulin sensitivity are the euglycaemic hyperinsulinaemic clamp and the intravenous glucose tolerance test with minimal model analysis. Insulin resistance occurs in many aetiologically diverse human disorders. Genetic syndromes with extreme insulin resistance are very uncommon. Insulin resistance is frequently associated with obesity, type 2 diabetes and essential hypertension. The insulin resistance syndrome called syndrome X includes impaired insulin-mediated glucose uptake, impaired glucose tolerance, hyperinsulinaemia, hypertension, dyslipidaemia and haemostatic disorders. Finally, the clinical significance of high values of insulin sensitivity is discussed. PMID- 9754277 TI - [Anti-HCV serology for screening, diagnosis and surveillance of hepatitis C: role of the immunoblot]. AB - Screening for antibody to hepatitis C virus (anti-HCV) is usually carried out using microplate enzyme immunoassays (EIA). According to French Ministry of Health recommendations, any positive or dubious result has to be controlled on a second blood sample with a anti-HCV assay differing from the one used for the initial screening. This second test can either be another EIA or an immunoblot assay. The present article investigates the advantage of a strip immunoblot assay including 4-viral peptides (Riba-3) for this control. The use of Riba for the early diagnosis of hepatitis C infection and for the follow-up of infected patients is also evaluated. Riba can be used to assess false positive EIA reactions, for instance in cases with isolated antibody to NS5. From large clinical series, it appears that positive Riba serum profiles can be divided in two main groups: Strongly positive sera associate at least strong (3 or 4+) anti capsid and anti-NS3 antibody reactivities. These profiles are often completed by anti-NS4 and/or anti-NS5 reactivities. Such profiles are associated with HCV viremia in about 90% of the cases. In this group, prevalence of viremia reaches 100% in patients with elevated level of serum alanin aminotransferase (ALT) and is slightly less frequent, 85%, in patients with normal level of ALT; Weakly positive sera lack either anti-capsid or anti-NS3 antibodies or exhibit only low reactivities (1+ or 2+) to these two antigens. These profiles can also be associated with antibodies to NS4 and/or NS5. Only 10% of patients with such Riba profiles are viremic. Weakly positive profile can be encountered during the early phase of HCV infection (recent seroconversion) or in patients who experience a favourable evolution of their HCV infection. Change of Riba profiles during follow-up provides fruitful information on recent seroconversions (increase in number and intensity of the reactivities). It can also help predict the outcome of HCV infection, a diminution of the intensity of Riba reactivities will confirm the extinction of viral replication. PMID- 9754278 TI - [MSRV retrovirus and gliotoxin protein: potential biological markers in multiple sclerosis?]. AB - The aetiopathogeny of multiple sclerosis, a neurological disease which prevalence and duration generate an important problem of public health in industrialized countries where it is most frequent, is not clearly understood. The keys for the diagnosis and therapeutic strategies of MS are nonetheless dependent upon the identification of a well-defined aetiology and upon the understanding of the mechanisms and pathogenic connexions which lead to the demyelinating lesions of the central nervous system and to the dysfunctions of the immune system (autoimmunity) characteristic for the disease. The recent identification of a retroviral agent (MSRV) which produces extracellular particles detectable in the plasma, the CSF, and in some cell cultures from patients with MS, as well as of a cytotoxic factor targeting glial cells (gliotoxin) detected in parallel, could help elucidating the aetiopathogeny of MS. The usefulness of biological markers and targets derived from MSRV retrovirus and this gliotoxin, in the diagnostic and therapeutic perspectives for MS, is discussed in the light of the different aetiopathogenic hypotheses for the disease. From our results it is conceivable that a retroviral agent and pathogenic molecules such as this gliotoxin and an eventual MSRV-associated retroviral superantigen might initiate and perpetuate the cascade of events leading to MS. However, similar data on different simple retroviruses were recently published concerning autoimmune diseases such as diabetes type 1, Sjogren's syndrome and systemic lupus erythematosis, which could prefigurate a broader concept for the role of such retroviruses in the aetiopathogenesis of autoimmune diseases. PMID- 9754279 TI - [Unusual behavior of serum markers in risk evaluation for trisomy 21]. AB - Screening of Down syndrome using serum markers is based on statistic risk determination calculated from the results of markers. An increased risk of fetal Down syndrome is associated with high hCG levels and low AFP levels in maternal serum. In the daily practice, the use of these two markers also leads to observation of different analytical patterns. We reviewed these patterns according to published data and our own experience. Some patterns are well documented (neural tube defects or trisomy 18) some of them remain unexplored. Numerous difficulties are encountered in the clinical use of these markers patterns: lack of consensus for their analytical definition, problems in interpretation, lack of regular dispositions. PMID- 9754280 TI - [Comparison of three reactants for the detection of activated protein C resistance due to mutation of factor V Leiden during pregnancy]. AB - The presence of the R506Q mutation of the factor V gene is associated with an increased risk of thromboembolism, particularly during pregnancy. Recently, its involvement in the development of obstetrical complications, such as preeclampsia and fetal losses, has been evoked. The resulting factor VQ506 (factor V Leiden) has arginine 506 replaced by glutamine at the factor Va cleavage site for activated protein C (APC) which induces APC-resistance. During pregnancy, an acquired resistance to APC is observed without the presence of the factor V Leiden mutation which leads to an inappropriate realization of the more expensive DNA analysis. This resistance is at least partly explained by an increase of the factor VIII. In this study, we have compared three reagents: the original test Coatest APC Resistance (Chromogenix) and two modified tests using factor V depleted plasma: Coatest APC Resistance V (Chromogenix) and Accelerimat (BioMerieux). The last test is not influenced by the factor VIII by the adjunction of activated factor X. For each test, the coefficient of discrimination, between carrier and non-carrier of the R506Q mutation of the factor V gene, has been determined on 43 pregnant women (33 non-carriers and 11 heterozygotes) and 51 unselected non pregnant patients with clinically suspected thrombosis (40 non-carriers and 11 heterozygotes). The predilution of the patient's plasma with factor V deficient plasma (Coatest APC Resistance V and Accelerimat) enhances the discrimination between carriers and non-carriers in both groups. However, using Coatest APC Resistance V, a significant difference of results is observed between the two populations in the non-carriers patients. Thus, Accelerimat is probably more efficient than Coatest APC Resistance for the detection of the factor VQ506 during pregnancy. PMID- 9754281 TI - [Plasma apolipoproteins AI and B: reference values in the Isere population and determination of individual values]. AB - Reference ranges for apolipoprotein AI and B plasma concentrations were established in a population of unrelated apparently healthy volunteers (138 men and 186 women) living in the region of Grenoble. Apolipoproteins were measured using an immunoturbidimetric assay on a Cobas Fara II analyzer, with reagents and IFCC standardized calibrators from Orion. Apolipoprotein AI mean concentration was higher in women than in men and increased with age in both men and women older than 45. Apolipoprotein B mean concentration was higher in men and increased linearly with age in both sexes. Linear regression analysis was used to determine desirable and high risk values for apolipoproteins AI and B from the guidelines developed by the National Cholesterol Education Program for HDL cholesterol and LDL cholesterol, respectively. Our data indicate that an apolipoprotein AI value of 1.05 g/l is comparable to an HDL cholesterol value of 0.35 g/l. The apolipoprotein B cutpoints of 1.15 g/l for men and 1.05 g/l for women correspond to the accepted LDL cholesterol cutpoint of 1.60 g/l. PMID- 9754282 TI - [Lipoprotein(a) and other lipid parameters in cord blood: a study of 528 cases]. AB - We have studied the concentrations of cholesterol, triglycerides, high-density lipoprotein cholesterol, apolipoproteins A-I and B, and lipoprotein (a) in cord blood, from children born in our hospital for a five-month period. Cholesterol and triglycerides values were slightly lower than values obtained in cord blood within other populations. The distribution of lipoprotein (a) concentrations was markedly skewed towards low values (mean = 1.74 mg/dl, median = 1 mg/dl), similar to that of other young or adult Caucasian populations. Children with a positive familial history of cardiovascular heart disease had a higher mean lipoprotein level (a), and a higher prevalence of lipoprotein values exceeding 5 mg/dl, than children with a negative familial history. These results suggest that lipoprotein (a) is an important risk factor for cardiovascular heart disease. PMID- 9754283 TI - [Diabetes mellitus: proposal of new diagnostic and classification criteria]. PMID- 9754284 TI - [Fetal trisomy 21 risk evaluation by measurement of maternal serum markers]. PMID- 9754285 TI - [Co-infection with Cryptosporidium sp and Cyclospora sp in an AIDS stage HIV patient]. PMID- 9754286 TI - [Troponin I, total CKMB: which marker to trust?]. PMID- 9754287 TI - [Postinfection encephalitis: a previously unrecognized complication of measles]. PMID- 9754288 TI - [Monoclonal IGM interference during measurement of reactive protein C and ferritin by immunonephelometry]. PMID- 9754289 TI - ["Point-of-care" or "point-of-need"? Tentative definition of common technology]. PMID- 9754290 TI - [Survey of satellite laboratory use versus analytical laboratories in health institutions]. PMID- 9754291 TI - [Evaluation of the quality control system in medical biology]. PMID- 9754292 TI - [The genus Ctenocephalides Stiles and Collins, 1930 (Siphonaptera, Pulicidae)]. AB - After a short historical review of this genus and an outline of biogeographical and paleontological origin of this flea, the 14 taxa are studied (synonymy, repartition, specificity and morphology). A new dichotomic key is given on criteria selected by the authors. PMID- 9754293 TI - Schellackia calotesi n. sp. from agamid lizards of the genus Calotes in Thailand. AB - Schellackia calotesi n. sp. is described from the Thai agamids Calotes mystaceus and C. versicolor. Schellackia-type sporozoites were recovered from blood and liver of one C. versicolor from Kon Kaen North-East Thailand and two C. mystaceus from Chiang Mai, North Thailand. Specimens of both species were fed on sporozoite infected blood, of these only one C. mystaceus developed endogenous infection in the anterior intestine. Description, from histological material includes early and dividing meronts, micro and macrogamonts and non sporulated oocysts. PMID- 9754294 TI - The in vitro effects of crystal violet on the pathogenic haemoflagellate Cryptobia salmositica Katz, 1951 (Sarcomastigophora: Kinetoplastida). AB - Crystal violet does not inhibit in vitro multiplication of a nonpathogenic strain of Cryptobia salmositica at low concentrations (0.01 microM and 0.001 microM) but multiplication is inhibited at higher concentrations (> or = 0.05 microM). In contrast, the pathogenic strain of C. salmositica does not multiply in vitro when incubated with crystal violet (0.001 microM, 0.01 microM and 0.05 microM). The infectivity of the pathogenic strain is significantly reduced after in vitro exposure to crystal violet. Crystal violet lyses C. salmositica (100.0 microM) and causes lesions on mitochondrial and nuclear membranes of the parasite. Pathogenic strains of Cryptobia salmositica and C. bullocki are more susceptible to lysis after in vitro exposure to crystal violet than are nonpathogenic strains of Cryptobia salmositica and C. catostomi. PMID- 9754295 TI - New features on the moults and morphogenesis of the human filaria Loa loa by using rodent hosts consequences. AB - The development of the human filaria Loa loa (Dirofilariinae, Onchocercidae), previously studied in monkeys, was studied using the non permissive hosts-mice and jirds. The development proved to be rapid: moult 3 occurred on day 8 post inoculation, the adult stage was reached on day 25 and measured at that time 3 3.5 mm in length. As in the other filarioids, the female genital apparatus developed during the fourth stage. A critical analysis of the studies on the development of Onchocercid species was made. The optimal duration of the stages (i.g. the shortest time) was chosen for the comparison. It appeared that the duration of the stage 3 was a constant character in a given species whatever the experimental conditions, whereas moult 4 might be retarded in a non susceptible host. Comparison between the 18 developmental cycles of Onchocercidae in the vertebrate host was made. Two biological types could be distinguished: either the moult 3 occurred on day 2-3 and was followed apparently by a late moult 4 (> or = 50 days), or the moult 3 occurred after about one week of development and it was associated with a less long stage 4 (20-40 days). The first group includes Dirofilaria and Onchocerca, the second group brings together mainly Loa and the Onchocercinae of the Dipetalonema line and related genera (Acanthocheilonema, Brugia, Litomosoides, etc.). The groups thus formed suggest real relationships as they fit with the morphology of the infective stage and the results of a recent molecular analysis of the 5S DNA. PMID- 9754296 TI - Use of random amplified polymorphic DNA (RAPD) for generating specific DNA probes for oxyuroid species (Nematoda). AB - Random amplified DNA markers (RAPD; Williams et al., 1990) were used to obtained specific RAPD fragments characterising different species of oxyuroids. We tested six species of worms parasitizing vertebrates or invertebrates: Passalurus ambiguus Rudolphi, 1819, parasite of Leporids; Syphacia obvelata (Rudolphi, 1802) Seurat, 1916, a parasite of rodents; Blatticola blattae (Graeffe, 1860) Chitwood, 1932 parasite of the cockroach Blattella germanica; Hammerschmidtiella diesingi (Hammerschmidt, 1838) Chitwood, 1932 and Thelastoma bulhoesi (Magalhaes, 1990) Travassos, 1929, parasites of the cockroach Periplaneta americana, and an undescribed parasite species of a passalid insect from New Caledonia. Among 15 oligonucleotides tested, nine produced several specific bands allowing the interspecific discrimination. PMID- 9754297 TI - [Interepidemic supervision of the leishmania focus of Keur Moussa (Thies, Senegal)]. AB - A supervision of the focus of human cutaneous leishmaniasis of Keur Moussa has been carried out in 1988-1989 and in 1991-1992. Among the 13 species gathering the 10,144 phlebotomine sandflies trapped, two belong to genus Phlebotomus, the others to genus Sergentomyia. Phlebotomus duboscqi, that has been found out by Deded et al. in 1980 as the vector of this leishmaniasis in Senegal, is the most represented species after Sergentomyia schwetzi, with respectively 32.3 and 28.5% of the found during these two periods. It is twice more abundant and frequent in the monastery area than the religious one. This difference may be due to the better micro-climatic conditions, the abundance of rodents and the human proximity. After the rainy seasons that influence a lot P. duboscqi's activity, the density of phlebotomine sandflies makes a progressive rise, as well as the temperature and humidity, to reach two maxima in April-May and July-August. Parasitological studies on females of phlebotomine sandflies and rodents Arvicanthis niloticus, Mastomys erythroleucus and Cricetomys gambianus are negative. This focus seems to be again in an inter-epidemic phase. PMID- 9754298 TI - [Estimation of the prevalence of bovine hydatid cyst in the south Pyrenees]. AB - Since 1994 "Reseau VEGA" (veterinary survey network) has organized a record of sanitary information in 14 slaughterhouses in the Midi-Pyrenees. Data about hydatidosis in cattle are centralized, analysed, then sent namely to each stockbreeder concerned. Estimation of the prevalence rate from 1994 to 1996 is 0.28% for animals and 2.5% for livestock. A marked decrease of rates was noticed during this three year monitoring period. Nevertheless, the Pyrenean area remains more affected than the North of the region. In an outbreak of hydatidosis, a few animals are carriers. Bovine infestation must be considered as revealing a rural cycle. Moreover, the link between bovine hydatidosis and ovine transhumance seems to be confirmed. Using livestock as epidemiological units is innovative in terms of hydatidosis. This approach allows a better adjustment of parasitism control and introduces the notions of the outbreak and the risk of human contamination. In the Midi-Pyrenees region, local human cases of hydatidosis are few. However, the absence of compulsory notification and of databases, on the one hand, and the extreme difficulty of confirming the autochthonous nature of the contamination, on the other hand, limit the reliability of data. A better collaboration between physicians and veterinarians would reduce animal prevalence and the risk of human contamination. PMID- 9754299 TI - Comparison of effects of low and high tick infestations on acquired cattle tick resistance: Hyalomma marginatum marginatum. AB - Three Holstein calves were infested with low numbers of ticks, two or three pairs of adults Hyalomma marginatum marginatum in cloth bag daily for 21 days. Infestation was carried out during tick proliferation periods. Two months later, cattle leads were challenged with 100 pairs of ticks. Another group of three Holstein calves were infested five times with 100 pairs of adult ticks of the same species. The five infestations were performed two weeks from the previous infestation. Three tick characteristics were recorded for each experiment: survival to detachment, females weight at detachment and egg mass weight. Light continuous infestations did not cause a significant change in this parameter, but every parameter declined gradually in the heavy infestations. Female and egg mass weight reached a significant difference from the first infestation by the fourth infestation. The circulating antibodies anti-salivary glands of Hyalomma m. marginatum showed that light infestation may induce like immuno-suppression. However, there is an inverse relationship between these antibodies and manifestation of resistance when calves were heavily infested. This is discussed in relation to a fraction of produced antibodies against protective antigens, and participation of another effector mechanism. PMID- 9754300 TI - Chitinolytic activities in Trichomonas vaginalis. AB - Chitinolytic activities were identified in the Protozoa Trichomonas vaginalis. Overall chitinase activity assessed using chitine-azure as substrate was 10.93 +/ 1.21 nmoles/min/mg prot. End nonreducing chitobiosidase (exochitinase) and chitotriosidase (endochitinase) activities were shown using p-nitrophenyl substrates and had specific activities of 4.55 +/- 0.53 and 0.47 +/- 0.06 nmol/min/mg prot, Kmapp. = 1.32 mM and 5 microM and pH optimum = 7.0 and 6.1 respectively. beta-N-acetylhexosaminidase (NAHase) activity was also detected with a specific activity of 5.40 +/- 0.65 nmol/min/mg prot., Kmapp = 0.656 mM and pH optimum at 7.0. No release of these enzymes into the culture medium was found. The possible role of chitinases in T. vaginalis remains to be investigated. PMID- 9754301 TI - Hemolytic activity of Monocercomonas spp. AB - The hemolytic activity of an isolate of Monocercomonas spp. from Tropidophis melanurus (snake: Boidae) was investigated. The isolate was tested against human erythrocytes of groups A, B, AB and O and against erythrocytes of six adult animals of different species (rabbit, rat, chicken, horse, bovine, and sheep). Results show that Monocercomonas spp. exerted an hemolytic activity against all erythrocytes tested. PMID- 9754302 TI - Antifeeding effect of several insecticidal formulations against Ctenocephalides felis on cats. AB - Evaluation of insecticidal activity of flea products is generally based on counting live fleas in the animal's coat 24 and 48 hours following artificial infestation. This approach, however, does not allow to specify whether the fleas have had the opportunity to bite and take a bloodmeal prior to their death. To address this question, 30 cats were alloted to six groups of five animals. Each cat was housed in a separate cage. At Day 0, each group of cats received a single treatment as follows: Group 1: spot-on application of imidacloprid: cats < 4 kg: 40 mg/cat, cats > or = 4 kg: 80 mg/cat (Advantage). Group 2: spot-on application of fipronil: 50 mg/cat (Frontline spot-on). Group 3: spray application of fipronil: 7.5 mg/kg b.w. (Frontline spray). Group 4: foam application of permethrin 40/60: 50 mg/kg b.w. (Defencat). Group 5: aerosol spray application of dichlorvos + fenitrothion: one second/kg b.w. (NuvanTop). Group 6: control group: cats were left untreated. One hour after treatment, each cat was infested with 50 unengorged young adult fleas, Ctenocephalides felis, deposited along the dorsal midline. One hour later, each cat was carefully combed using a fine-toothed comb (13 teeth/cm). Collected fleas were swatted to deteci blood in their abdomen. To the manufacturers respective product use instructions and efficacy claims, reeinfestations were made at Days 3, 7, 14 in all groups; at Days 21 and 30 in Groups 1, 2, 4, 6; at Days 35 and 42 in Groups 3 and 6. The cats were combed one hour after each reinfestation. The results indicate that the topical application of imidacloprid or fipronil does not prevent fleas from biting and feeding within the first hour after infestation prior to being killed while the topical application of dichlorvos/fenitrothion and permethrin let to a better than 80% decrease of the number of engorged fleas for three and seven days post treatment, respectively. PMID- 9754303 TI - Age grading Anopheles arabiensis: their gorging and surviving responses using a membrane feeding system. AB - Literature on artificial membrane feeding has traditionally supported the notion that mosquitoes have to be enough old to present a high gorging rate and enough young to present a high surviving rate. In order to know the best mosquito age to perform a Plasmodium sp. experimental infection revealed by oocyst examination, Anopheles arabiensis of known ages were fed on baudruche membrane with fresh human blood thermostated at 37 degrees C. The response was evaluated in term of gorging after ten minutes in contact with the membrane feeder and, for engorged females, in term of survival eight days after the engorgement. Two strains of mosquitoes were used, one wild, collected at larval stages in the city of Dakar and the other, originating from the same place but reared in our insectary since three years. The two strains showed their highest gorging rate at 3-4 days after emergence (39.4% for the wild strain and 63.3% for the insectary strain). The survival rate of the wild strain was highest at 2-3-4 days after emergence (64.8%) and those of the insectary strain at four days (65.6%). In conclusion, the gorging mosquitoes would have to be aged of 3-4 days after emergence for the wild strain and of four days for the insectary strain in order to obtain eight days after the engorging the best output of fed and surviving mosquitoes. PMID- 9754304 TI - False-positive Trichuris suis egg counts in pigs in relation to coprophagia. AB - Sixteen non-infected control pigs housed together with 16 pigs infected with Trichuris suis, excreted T. suis eggs in their faeces (range 20-4, 960 eggs per gram faeces (EPG)). When the control pigs were moved to clean pens, their egg counts dropped to zero EPG within five days. A significant correlation was found between T. suis egg counts of infected and control pigs penned together (r = 0.89, P < 0.001). These results suggest that false-positive faecal egg counts may be the result of coprophagia. PMID- 9754305 TI - Homology of Trypanosoma cruzi clone 36 repetitive DNA sequence to sequence encoding human Ro/SSA 52 kD autoantigen. PMID- 9754306 TI - [Cytoskeletal actin and its associated proteins. Some examples in Protista]. AB - Many processes, cell motility being an example, require cells to remodel the actin cytoskeleton in response to both intracellular and extracellular signals. Reorganization of the actin cytoskeleton involves the rapid disassembly and reassembly of actin filaments, a phenomenon regulated by the action of particular actin-binding proteins. In recent years, an interest in studying actin regulation in unicellular organisms has arisen. Parasitic protozoan are among these organisms and studies of the cytoskeleton functions of these protozoan are relevant related to either cell biology or pathogenicity. To discuss recent data in this field, a symposium concerning "Actin and actin-binding proteins in protists" was held on May 8-11 in Paris, France, during the XXXV meeting of the French Society of Protistology. As a brief summary of the symposium we report here findings concerning the in vitro actin dynamic assembly, as well as the characterization of several actin-binding proteins from the parasitic protozoan Entamoeba histolytica, Trichomonas vaginalis and Plasmodium knowlesi. In addition, localization of actin in non-pathogen protists such as Prorocentrum micans and Crypthecodinium cohnii is also presented. The data show that some actin-binding proteins facilitate organization of filaments into higher order structures as pseudopods, while others have regulatory functions, indicating very particular roles for actin-binding proteins. One of the proteins discussed during the symposium, the actin depolymerizing factor ADF, was shown to enhance the treadmilling rate of actin filaments. In vitro, ADF binds to the ADP-bound forms of G-actin and F-actin, thereby participating in and changing the rate of actin assembly. Biochemical approaches allowed the identification of a protein complex formed by HSP/C70-cap32-34 which might also be involved in depolymerization of F actin in P. knowlesi. Molecular and cellular approaches were used to identify proteins such as ABP-120 and myosin IB at the leading edge of E. histolytica. ABP 120 organizes F-actin in a network and myosin IB participates in the pseudopod formation. Similar approaches using T. vaginalis resulted in the discovery of an actin-binding protein that participate in the F-actin reorganization during adhesion of parasites to target cells. This protein is homologous to alpha actinin from other eukaryotic cells. Finally, by using cell biology approaches, F actin was observed in the cytoplasm as well as in the nucleus of Dinoflagellates. The recent developments in the molecular genetics of protozoa will provide new insights to understand the roles of actin-binding proteins during cytoskeleton activities. PMID- 9754307 TI - Cercopithifilaria shohoi n. sp. (Nematoda: Filarioidea) from the relict Bovidae, Capricornis crispus, in Japan. AB - Cercopithifilaria shohoi n. sp. was found in the relict bovid, Capricornis crispus, in Japan, and is described and compared with other species in the genus. Adult male and female worms were found in subcutaneous tissues of the trunks of 6 serows shot in Mt. Zao, Yamagata Prefecture, in the northern part of Honshu, the largest island of Japan. The one complete male found was 19.7 mm long, and the five females were 31.6-50.9 mm long. Unsheathed or sheathed microfilariae 104-122 microns long were taken from the females. One microfilaria was found in the sediment of the preservation solution of the tissues, but none were found in the blood of the infected serows, so microfilariae may be limited to the skin. Males of this species had one pair of papillae between perianal and subterminal groups of caudal papillae. In having this intermediate pair, C. shohoi n. sp. resembled species such as C. faini from an African bovid and C. rugosicauda from a European deer. From its morphological characteristics, C. shohoi n. sp. seems to be one of the more primitive species in the genus Cercopithifilaria. PMID- 9754308 TI - Larvae and adults of Hysterothylacium aduncum (Rudolphi, 1802) (Nematoda: Anisakidae) in fishes and crustaceans in the south west Atlantic. AB - Hysterothylacium aduncum (Rudolphi, 1802) is reported from five fishes and one invertebrate species. Third-stage larvae were found in the crustacean Themisto gaudichaudii and in mesenteries of the fishes Engraulis anchoita and Merluccius hubbsi; fourth-stage larvae were recovered from the digestive tract of M. hubbsi and Scomber japonicus and adult specimens were obtained from the stomach and intestine of M. hubbsi, S. japonicus, Genypterus blacodes and Genypterus brasiliensis. Nematodes are described, measured and illustrated. Parasitic prevalence, mean intensity and range were calculated in relation to different geographic zones, from the Argentinean-Uruguayan Common Fishing Zone to Patagonic areas. An increase of parasitism from the northern areas southwards was observed. The life-cycle of H. aduncum, involving the host species considered, is also postulated. PMID- 9754309 TI - Faecal egg counts are representative of digestive-tract strongyle worm burdens in sheep and goats. AB - The relationship between faecal egg counts and worm burdens in sheep and goats was studied in a large array of environments, from temperate (ewes, lambs or dairy-goats in France) or steppic (ewes in Middle-Atlas of Morocco) to Sahelian (young sheep and goats of Mauritania in West Africa) climates. The studied temperate worm communities were dominated by Teladorsagia and Trichostrongylus sp., and those from steppic areas by Teladorsagia, Marshallagia and Trichostrongylus sp.; Haemonchus contortus was highly predominant in the Sahelian regions. The fertility of worms depended on density (10 to 50% of variance) and presence of H. contortus to a lesser extent. For sheep and goats from several temperate and steppic areas, a good relationship between egg counts and worm burdens was established (r = 0.62). It was ameliorated when the percentage of H. contortus, the most prolific species was incorporated in the model. The predictive value of faecal egg count for assessing worm burden was only of interest for groups of hosts. PMID- 9754310 TI - [Phlebotomines of Senegal (Diptera: Psychodidae): population and population dynamics of the Mont-Rolland region]. AB - Phlebotomine sandflies were captured on a monthly basis from May 1995 to April 1996 in the Mont-Rolland district in Western Senegal. The objectives were to study the population dynamics of sandflies and to make an inventory of the viruses they transmit. Among 10,315 specimens captured, belonging to 14 species, Sergentomyia dubia (35.9%), S. schwetzi (27.7%) and S. buxtoni (24.5%) were the most abundant. Species from the genus Sergentomyia accounted for 99.6% versus 0.4% for the genus Phlebotomus. The sandflies population was observed to peak in February. The most populated resting sites of the captured insects were in decreasing order tree-holes, termite-hills and burrows. S. dubia was the most abundant species captured in tree-holes. It was S. buxtoni in termite-hills, while S. schwetzi was found to dwell most often in burrows. No virus was isolated from 2,114 specimens tested. PMID- 9754311 TI - [Description of Phlebotomus (Paraphlebotomus) riouxi n. sp. (Diptera:Psychodidae) of northern Africa]. AB - Description of a new palearctic species from Morocco, Tunisia and Spain: Phlebotomus (Paraphlebotomus) riouxi closely related to P. chabaudi. Differential identification with the neighbouring species. The aedeagus of the male is unrecurved, a character which distinguishes P. riouxi from other species with recurved aedeagus and the basal process of the coxite is bigger and more tufted than in P. chabaudi. For female, which has the same spermatheca with a typical terminal ring, the only available character to differentiate from P. chabaudi is the aspect of the armature in the genital atrium. PMID- 9754312 TI - [Vectorial competence of Glossina palpalis palpalis, Glossina p. gambiensis and Glossina morsitans morsitans flies for a clone of Trypanosoma (Nannomonas) congolense IL 1180]. AB - The authors report on the results of experimental infections of teneral (age < 32 hours) and non-teneral (age between 80 and 96 hours) Glossina palpalis palpalis, G. p. gambiensis and G. morsitans morsitans with Trypanosoma congolense IL 1180. Flies were infected once on a parasitaemic rat. Teneral flies, both sexes indiscriminate, showed a procyclic and metacyclic infection rate respectively of 0.0588 and 0.7272 for G. p. palpalis; 0.0525 and 0.0416 for G. p. gambiensis; 0.6493 and 0.7300 for G. m. morsitans. Neither of the non-teneral G. palpalis subspecies had any vectorial competence, whereas G. m. morsitans had procyclic and metacyclic infection rates of 0.4541 and 0.7884. Statistical analysis could not demonstrate any significant difference in metacyclic infection rate between teneral and non-teneral G. m. morsitans. Teneral flies of each subspecies transmitted the infection to rats, used as hosts, before the twentieth day. Concerning trypanosome development in the fly, it was observed that five days after infection procyclic and mesocyclic forms could be observed simultaneously in all flies dissected at that moment. PMID- 9754313 TI - Development and characterization of paromomycin-resistant Leishmania donovani promastigotes. AB - Paromomycin is an antileishmanial chemotherapeutic agent. Leishmania donovani promastigotes resistant to 800 microM of paromomycin were selected by increasing drug pressure and cloned. These promastigotes did not acquire multidrug resistance. Paromomycin resistance was stable in the absence of the drug in the culture. It remained stable also in amastigotes isolated after a passage in mice. Furthermore the resistant parasites were still infective to macrophages in vitro and for mice in vivo. A sensitive method to detect and to quantify intracellular paromomycin by HPLC was developed and allowed to show that the main mechanism of resistance seems to be due to decreased drug uptake probably as a consequence of altered membrane composition. PMID- 9754314 TI - N-methylglucamine antimonate (SbV+): intralesional canine tegumentary leishmaniasis therapy. AB - Twenty five adult dogs of three municipalities of the State of Rio de Janeiro, Brazil, that had been naturally infected by L. (V) braziliensis were treated with N-methylglucamine antimonate (Glucantime). Nine of the animals (36.0%) presented ulcerated skin lesions, twelve (48.0%) had mucosal lesions and four (16.0%) had multiple lesions. In some cases the mucosal lesions were associated to skin lesions. A dose of 85 mg SbV+ or 1 ml of the drug was intralesionally administered to the dogs. The animals were divided into three groups according to the amount of necessary doses (between one and three) for the complete healing of the lesions. The dogs were observed for six months after the third group received the last dose. Within this period two animals perished. Serial antibody evaluation through IFAT has shown that in 14 samples (63.3%) the titers have remained unaltered, in four of them (16%) there has been a decrease in two titers and in five of them (21%) serology was negative. Nineteen of the dogs (86.6%) had their lesions completely healed. The authors suggest intralesional therapy be the first choice of treatment of canine tegumentary leishmaniasis due to its effectiveness. PMID- 9754315 TI - Effect of the infection by Neostrongylus linearis on the survival of the intermediate host Cernuella (Cernuella) virgata. AB - The molluse intermediary host Cernuella (Cernuella) virgata was infected both in nature and in the laboratory with Neostrongylus linearis larvae. The infection rate was higher in the natural infection than in the experimental one all through the study. Accordingly, the survival of the molluscs was lower among those that had been naturally infected than among uninfected control and experimentally infected ones. The differences between survival curves of the different batches were only significant in the months when the infection rate in naturally infected batches was very high, from November to February. So we can conclude that the mortality rate is higher among the snails which harbour a considerable number of N. linearis than among non-infected or moderately infected molluscs. PMID- 9754316 TI - Argentophilic structures of the miracidium of Echinochasmus perfoliatus (Trematoda: Echinostomatidae). AB - Argentophilic structures of the miracidium of Echinochasmus perfoliatus were described from material collected in the vicinity of Vladivostok, Far East of Russia. Impregnated with 0.5% solution of AgNO3 miracidium showed 21 epidermal plates arranged in four rows: 6 + 9 + 4 + 2. Up to 23 papilla-like structures on the terebratorium were arranged along three axes and in four groups. A single papilla was located at the base of each of ventral and dorsal epidermal plates of the first row. Two papillae were located at the base of each of lateral epidermal plates of the first row. The eyespots were located posterior to the first row of plates. Two excretory pores were located anterior to the last row of plates. The results obtained were compared with the argentophilic structures of closely related species of the genus Echinochasmus. PMID- 9754317 TI - Diagnosis of Plasmodium falciparum malaria using ParaSight F, ICT malaria PF and malaria IgG CELISA assays. AB - A battery of sixty-six blood samples from Senegal was analysed by the ParaSight F test, the ICT Malaria PF and the Malaria IgG CELISA. These three assays detect the histidine rich protein 2 antigen of Plasmodium falciparum. Thick smear microscopy was used as the reference method. Sensitivity, specificity, predictive positive and negative values were respectively 89%, 100%, 100%, 88% for the ICT; 86%, 93%, 94%, 85% for the paraSight and 88%, 87%, 88%, 87% for the Malaria IgG CELISA. The three assays failed to detect two positive samples with P. ovale and P. malariae. Assays were also compared with regard to the expense of equipment and reagents and speed and ease of use. The rapid ICT and ParaSight F test can be performed with minimal training and may be specially useful in areas where P. falciparum is the predominant malaria species, in epidemic malaria regions, and where skilled microscopy is not readily available. PMID- 9754318 TI - Recent development in control of Theileria annulata in Iran. AB - During 1973-1990 cattle immunization, in Iran, was induced by two strains within an interval of one month: first the milder and then the mild strain. Although this method of vaccination rendered satisfactory results in the field, yet production, maintenance in deep-freezers and transportation in liquid nitrogen particularly to remote areas of the country proved to be uneconomical and time consuming. Therefore, in order to reduce cost and save time, a new method involving only one local and live attenuated vaccine strain was sought. Reports received from different ecological areas of the country have shown no presence of any significant abnormal side-effects in vaccinated cattle and the immunization results have been highly satisfactory. PMID- 9754319 TI - Faunal composition and behavior of anopheline mosquitoes in the Xavante Indian reservation of Pimentel Barbosa, central Brazil. AB - Faunal composition and behavior of anopheline mosquitoes were studied in a Xavante Indian reservation of Central Brazil. Altogether 558 anophelines were collected in three environments (intra, peri, and extra-domiciliary). Anopheles darlingi (30.9%), An. triannulatus s.l. (24.6%) and An. oswaldoi (19.7%) were the most common species. Average capture rates were higher in the rainy season (8.03 per hour) than in the dry season (4.37 per hour). Anophelines exhibited exophilic behavior almost exclusively. It was observed that Xavante cultural practices facilitate outdoor exposure during peak hours of mosquito activities (e.g., coming to the creek early in the morning for bathing or to draw water, fishing, etc.). The results of this study raise the question of whether or not applying to the Xavante the more commonly recommended malaria control strategies (e.g., in house spraying, screening windows, and impregnated bed nets) which aim at hampering human-mosquito contact inside human dwellings may be effective. PMID- 9754320 TI - Identification of virus-like particles in Trypanosoma cruzi by fine structure analysis. PMID- 9754321 TI - Molecular mechanism of the sensing of the hydrogen peroxide stress response in Escherichia coli. PMID- 9754322 TI - Peroxynitrite and myeloperoxidase leave the same footprint in protein nitration. PMID- 9754323 TI - Vitamin E may slow kidney failure owing to oxidative stress. AB - Progression to kidney failure in a number of major renal diseases is now thought to be significantly worsened by oxidative stress at the biochemical level. Evidence is accumulating that the rate of deterioration could, in many cases, be slowed down to a more acceptable level by the simple expedient of dietary supplementation with the antioxidant, vitamin E. Evidence for the potential use of vitamin E as an adjunctive therapy to help prolong kidney function in conditions that are accelerated by oxidative stress is discussed. PMID- 9754324 TI - Modification of red cell membrane lipids by hypochlorous acid and haemolysis by preformed lipid chlorohydrins. AB - Hypochlorous acid (HOCl), a strong oxidant generated by the myeloperoxidase system of neutrophils and monocytes, has been implicated in inflammatory tissue damage by these cells. Reaction of HOCl with the double bonds of unsaturated lipids produces alpha, beta-chlorohydrin isomers. We have exposed red cell membranes to HOCl and used thin layer chromatography (TLC) of the extracted lipids and enzyme-linked immunosorbent assay (ELISA), using an antichlorohydrin monoclonal antibody, to show that fatty acyl chlorohydrins are formed. The ELISA was approximately 25 fold more sensitive than TLC, and chlorohydrins were detected when membranes from 10(6) cells were treated with > or = 0.16 nmoles HOCl. Lipid chlorohydrins are more polar and bulky than their parent lipids and as such could affect membrane stability and function. To determine the effect of incorporation of lipid chlorohydrins into cell membranes, preformed fatty acid and cholesterol chlorohydrins were incubated with red cells. Lysis was measured as release of haemoglobin and incorporation of lipids was determined by 14C scintillation counting. Addition of HOCl-treated oleic acid to red cells resulted in rapid lysis of a fraction of the cells in a concentration dependent manner. HOCl-treated cholesterol also caused a small amount of cell lysis that was predominantly due to chlorohydrin 3, one of the three major cholesterol chlorohydrin products. Chlorohydrin 3, which has a decreased planarity and polarity, was also primarily responsible for altering the critical micelle concentration of HOCl-treated cholesterol-containing liposomes. PMID- 9754325 TI - Tyrosine hydroxylase: mechanisms of oxygen radical formation. AB - A new mechanism of oxygen radical formation in dopaminergic neurons is proposed, based on the oxidative mechanism of tyrosine hydroxylase. The cofactor (6R,6S) 5,6,7,8-tetrahydrobiopterin can rearrange in solution which allows an autoxidation reaction producing O2.-, H2O2 and HO.. The combination of tyrosine hydroxylase and the cofactor produces more oxygen radicals than does the autoxidation of the cofactor. This production of oxygen radicals could be damaging to dopaminergic neurons. In the presence of tyrosine, the enzyme produces less radicals than it does in the absence of tyrosine. Mechanisms are proposed for the generation of reactive oxygen species during the autoxidation of the cofactor and during enzymatic catalysis. The generation, by tyrosine hydroxylase, of very small amounts of oxygen radicals over the period of 65 years could contribute to the oxidative stress that causes Parkinson's disease. PMID- 9754326 TI - Phospholipid fatty acid composition affects enzymatic antioxidant defenses in cultured Swiss 3T3 fibroblasts. AB - The aim of this work was to study the adaptation of enzymatic antioxidant cell defense to the nature of the membrane polyunsaturated fatty acids (PUFA). 3T3 Swiss fibroblasts were grown for 5 days in a medium supplemented with 50 microM linoleic acid (LA) or eicosapentaenoic acid (EPA) and compared to control cells (C). The phospholipid fatty acid content was evaluated: LA were enriched in n-6 PUFA (27.8%) in comparison to C (6.7%) or EPA (5.6%); EPA were enriched in n-3 PUFA (26.2%) in comparison to LA (4.4%) or C (4.6%). The fatty acid double bond index (DBI) increased from C to LA and EPA. The activities of the three key enzymatic antioxidant defenses, SOD, GPx and GST, increased with the degree of unsaturation of the phospholipid fatty acids. In the cells with fatty acids that are very sensitive to oxidative stress, the higher activities of SOD and GPx might act to limit the initiation of lipid peroxidation and the higher activities of GST and GPx to decrease the toxic effects of the various species produced from lipid degradation. PMID- 9754327 TI - Ceruloplasmin releases pH-induced inhibition of cell proliferation stimulated by growth factors. AB - Swiss 3T3 fibroblasts can be weakly stimulated to grow by bombesin, epidermal growth factor or ceruloplasmin when cells are maintained in Dulbecco's Modified Essential Medium (DMEM), the pH of which is 7.75. Addition of insulin synergizes with the other mitogens. However, only ceruloplasmin promotes DNA synthesis in Minimum Essential Medium (MEM). The pH in this medium is 7.0. All the other growth factors synergize with the ceruloplasmin effects, but such synergism is not evident with insulin. If the pH in MEM is increased to 7.25 or 7.75 by supplementation with HEPES or NaHCO3, respectively, the results are similar to those found in DMEM. Since the oxidation of iron is increased at alkaline pH, the reoxidation of iron at the cell surface may facilitate growth at alkaline pH. We propose that iron reoxidation is limiting for cell growth and that part of the ceruloplasmin effect is mediated by its action as a terminal oxidase for ferrous iron on the cell surface. Observations consistent with this explanation include: 1) combinations of insulin with bombesin or epidermal growth factors do not promote cell proliferation at pH 7.0; 2) fetal calf serum, which has ferroxidase activity, and ceruloplasmin plus or minus other growth factors stimulate cell proliferation at pH 7.0; and 3) alkaline pH also restores the mitogenic effect of growth factors. PMID- 9754328 TI - Increased formation of interstitial hydroxyl radical following myocardial ischemia: possible relationship to endogenous opioid peptides. AB - The effects of myocardial ischemia and reperfusion on interstitial hydroxyl radical production, in the left ventricular myocardium of anesthetized cats, were investigated. Ringer's solution containing salicylic acid was perfused through an implanted microdialysis probe. Hydroxyl radical production was evaluated as the 2,3 and 2,5 dihydroxybenzoic acid (DHBA) concentrations in the microdialysates by an on-line high performance liquid chromatography system. Myocardial ischemia for 60 min, induced by ligation of the left anterior descending coronary artery, significantly increased both 2,3 and 2,5 DHBA levels when compared with the sham operated cats. Naloxone (1 mg/kg, bolus, intravenous), an endogenous opioid peptide receptor antagonist, significantly suppressed the ischemia-induced production of hydroxyl radicals. Myocardial ischemia also induced cardiac arrhythmia. Naloxone reduced the severity of ischemia-induced arrhythmia, as observed by a significantly lower arrhythmia score (1.4 +/- 0.2 vs. 4.6 +/- 0.4 for control), and by diminished incidence of ventricular tachycardia (0/7 vs. 8/8 for control) and ventricular fibrillation (0/7 vs. 3/8 for control). Furthermore, perfusion of dynorphin (0.25 microgram, 2.5 micrograms and 25 micrograms), an endogenous opioid peptide receptor agonist, increased hydroxyl radical production. Our results suggest that, in anesthetized cats, myocardial ischemia can induce production of interstitial hydroxyl radical in left ventricular myocardium, and this production may involve the actions of released endogenous opioid peptides on their receptors. PMID- 9754329 TI - Evidence that iron-overload promotes 7,12-dimethylbenz(a)anthracene- induced skin tumorigenesis in mice. AB - Iron overload is known to occur in West European and American populations due to the consumption of an iron-rich diet. There are also genetic disorders which lead to body iron overload. It has been shown that iron overload predisposes humans to an increased risk of cancer. In experimental animals, iron overload is known to enhance intestinal, colon, hepatic, pulmonary and mammary carcinogenesis. However, the mechanism by which iron overload enhances chemically-induced carcinogenesis is not known. In this study, we show that iron overload acts as a mild tumor promoter in mouse skin. Female albino swiss mice were given 1 mg iron/mouse parenterally for 2 weeks to induce iron overload. These animals showed a three-fold increase in cutaneous iron concentration as compared to normal mice. Tumors were initiated by topically applying 7,12-dimethylbenz(a)anthracene (DMBA). Appearance of the first tumor (latency period), percent tumor incidence and number of tumors/mouse were recorded. When compared to the control group, iron overload mice showed an increased incidence of tumors, from 25%-55% by week 20, and tumors appeared 4 weeks earlier. The number of tumors per mouse was four fold higher in the iron overload group. The induction of cutaneous ornithine decarboxylase (ODC) activity and [3H]thymidine incorporation in cutaneous DNA were higher in iron overload groups as compared to normal control animals. Similar to other oxidant tumor promoters, iron overload enhanced cutaneous lipid peroxidation and xanthine oxidase activity and decreased catalase activity. Our results indicate that iron overload exerts a mild tumor promoting activity in mouse skin. Our data also show that oxidative stress generated by iron overload plays an important role in the augmentation of cutaneous tumorigenesis. These data may also have implications for the enhanced risk of cancer-induction following UVB exposure of human populations with iron overload. PMID- 9754330 TI - Starvation-induced autophagocytosis paradoxically decreases the susceptibility to oxidative stress of the extremely oxidative stress-sensitive NIT insulinoma cells. AB - Glucose and amino acid starvation of cells in culture generally enhances their sensitivity to oxidative stress. This is explained by compensatory autophagocytosis, which results in increased amounts of lysosomal low-molecular weight, redox-active iron, due to the degradation of metallo-proteins, with a potential increase in iron-catalyzed, intralysosomal oxidative reactions. Such reactions diminish the stability of lysosomal membranes, with resultant leakage of hydrolytic enzymes into the cytosol and ensuing cellular degeneration, often of apoptotic type. However, starvation of NIT insulinoma cells, which are normally remarkably sensitive to oxidative stress, actually attenuated the sensitivity to such stress. We found that starved NIT cells rapidly synthesized ferritin. Moreover, ferritin was found to be autophagocytosed, and the lysosomes were stabilized, as assayed by the acridine orange relocation test. We hypothesize that compensatory autophagocytosis during starvation increases the cytosolic pool of redox-active iron, as a reflection of enhanced transportation of low-molecular-weight iron from autophagic lysosomes to the cytosol, resulting in ferritin induction. The newly formed ferritin would, in turn, become autophagocytosed and bind redox-active lysosomal iron in a non-redox-active form. We also suggest that the proposed mechanism may be a way for oxidative stress sensitive cells to compensate partly for their failing capacity to degrade hydrogen peroxide before it leaks into the acidic vacuolar apparatus and induces intralysosomal oxidative stress. The insulin-producing beta cell may belong to this type of cells. PMID- 9754332 TI - alpha-Tocopherol enhances the peroxidase activity of hemoglobin on phospholipid hydroperoxide. AB - We have used direct separation of phospholipid hydroperoxide and phospholipid hydroxide by high performance liquid chromatography to examine the phospholipid hydroperoxide peroxidase activity of hemoglobin (Hb) in the presence of hydrogen donors. Hb exhibits phospholipid hydroperoxide peroxidase activity and rapidly breaks down phospholipid hydroperoxide to thiobarbituric acid-reactive substances. However, in the presence of alpha-tocopherol, some phospholipid hydroperoxide is converted to phospholipid hydroxide, which is more stable than the hydroperoxide and is much less reactive with thiobarbituric acid. Other electron donors such as glutathione and ascorbate are less effective than alpha tocopherol. Free cysteine also shows some ability to reduce phospholipid hydroperoxides to corresponding hydroxides, but cys-93 beta of Hb did not participate in the reaction, as shown by N-ethylmaleimide modification. Hemin alone catalysed the reaction, in the absence of protein. The results therefore show that Hb catalyses an apparent phospholipid hydroperoxide alpha-tocopherol peroxidase reaction due to bound hemin, and that the reduction depends on the ability of hydrogen donors to react with the intermediate phospholipid alkoxyl radical and does not involve reduction by deprotonated sulfhydryl groups. PMID- 9754331 TI - Ferulic acid dehydrodimers from wheat bran: isolation, purification and antioxidant properties of 8-O-4-diferulic acid. AB - Wheat bran contains several ester-linked dehydrodimers of ferulic acid, which were detected and quantified after sequential alkaline hydrolysis. The major dimers released were: trans-5-[(E)-2-carboxyvinyl]-2-(4-hydroxy-3-methoxy-phenyl) 7-methoxy-2, 3- dihydrobenzofuran-3-carboxylic acid (5-8-BendiFA), (Z)-beta-[4 [(E)-2-carboxyvinyl]-2-methoxyphenoxy]-4-hydroxy-3-methox ycinnamic acid (8-O-4 diFA) and (E,E)-4,4'-dihydroxy-5,5'-dimethoxy-3,3'-bicinnamic acid (5-5-diFA). trans-7-hydroxy-1-(4-hydroxy-3methoxyphenyl)-6-methoxy-1,2-dihydro - naphthalene 2,3-dicarboxylic acid (8-8-diFA cyclic form) and 4,4'-dihydroxy-3,3'-dimethoxy beta,beta'-bicinnamic acid (8-8-diFA non cyclic form) were not detected. One of the most abundant dimers, 8-O-4-diFA, was purified from de-starched wheat bran after alkaline hydrolysis and preparative HPLC. The resultant product was identical to the chemically synthesised 8-O-4-dimer by TLC and HPLC as confirmed by 1H-NMR and mass spectrometry. The absorption maxima and absorption coefficients for the synthetic compound in ethanol were: lambda max: 323 nm, lambda min: 258 nm, epsilon lambda max (M-1 cm-1): 24,800 +/- 2100 and epsilon 280 (M-1 cm-1): 19,700 +/- 1100. The antioxidant properties of 8-O-4-diFA were assessed using: (a) inhibition of ascorbate/iron-induced peroxidation of phosphatidylcholine liposomes and; (b) scavenging of the radical cation of 2,2' azinobis (3-ethyl-benzothiazoline-6-sulphonate) (ABTS) relative to the water soluble vitamin E analogue, Trolox C. The 8-O-4-diFA was a better antioxidant than ferulic acid in both lipid and aqueous phases. This is the first report of the antioxidant activity of a natural diferulate obtained from a plant. PMID- 9754333 TI - t-Butylhydroperoxide and gliotoxin stimulate Ca2+ release from rat skeletal muscle mitochondria. AB - Rat liver mitochondria have a specific Ca2+ release pathway which operates when NAD+ is hydrolysed to nicotinamide and ADPribose. NAD+ hydrolysis is Ca(2+) dependent and inhibited by cyclosporine A (CSA). Mitochondrial Ca2+ release can be activated by the prooxidant t-butylhydroperoxide (tbh) or by gliotoxin (GT), a fungal metabolite of the epipolythiodioxopiperazine group. Tbh oxidizes NADH to NAD+ through an enzyme cascade consisting of glutathione peroxidase, glutathione reductase, and the energy linked transhydrogenase, whereas GT oxidizes some vicinal thiols to the disulfide form, a prerequisite for NAD+ hydrolysis. We report now that rat skeletal muscle mitochondria also contain a specific Ca2+ release pathway activated by both tbh and GT. Ca2+ release increases with the mitochondrial Ca2+ load, is completely inhibited in the presence of CSA, and is paralleled by pyridine nucleotide oxidation. In the presence of tbh and GT, mitochondria do not lose their membrane potential and do not swell, provided continuous release and re-uptake of Ca2+ ('Ca2+ cycling') is prevented. These data support the notion that both tbh- and GT-induced Ca2+ release are not the consequence of an unspecific increase of the inner membrane permeability ('pore' formation). Tbh induces Ca2+ release from rat skeletal muscle less efficiently than from liver mitochondria indicating that the coupling between tbh and NADH oxidation is much weaker in skeletal muscle mitochondria. This conclusion is corroborated by a much lower glutathione peroxidase activity in skeletal muscle than in liver mitochondria. The prooxidant-dependent pathway promotes, under drastic conditions (high mitochondrial Ca2+ loads and high tbh concentrations), Ca2+ release to about the same extent and rate as the Na+/Ca2+ exchanger. This renders the prooxidant-dependent pathway relevant in the pathophysiology of mitochondrial myopathies where its activation by an increased generation of reactive oxygen species probably results in excessive Ca2+ cycling and damage to mitochondria. PMID- 9754334 TI - Hydroxyl and 1-hydroxyethyl free radical detection using spin traps followed by derivatization and gas chromatography-mass spectrometry. AB - Detection of hydroxyl free radicals is frequently performed by electron spin resonance (ESR) following spin trapping of the radical using 5,5 dimethylpyrroline N-oxide (DMPO) to generate a stable free radical having a characteristic ESR spectrum. The necessary ESR equipment is expensive and not readily available to many laboratories. In the present study, a specific and sensitive gas chromatography-mass spectrometry (GC/MS) method for detection of hydroxyl and hydroxyethyl free radicals is described. The DMPO or N-t-butyl-alpha phenylnitrone (PBN) radical adducts are extracted and derivatized by trimethylsylilation and analyzed by GC/MS. To standardize the method, .OH and 1 hydroxyethyl radicals were generated in two different systems: 1) a Fenton reaction in a pure chemical system in the absence or presence of ethanol and 2) in liver microsomal suspensions where ethanol is metabolized in the presence of NADPH. In the Fenton system both radicals were easily detected and specifically identified using DMPO or PBN. In microsomal suspensions DMPO proved better for detection of .OH radicals and PBN more suitable for detection of 1-hydroxyethyl radicals. The procedure is specific, sensitive and potentially as useful as ESR. PMID- 9754335 TI - EPR detection of oxygen radical anion in aqueous solution at room temperature. PMID- 9754336 TI - -Cysts of the glenoid labrum-. AB - Cysts of the glenoid labrum are ganglia cysts extending near the glenohumeral joint with frequent clinical signs and symptoms of a compression of the subscapularis nerve. They are detected through MR examination. They are frequently associated to instability of the glenohumeral joint. PMID- 9754337 TI - -Arthrography and computed tomography arthrography of the wrist-. AB - After a brief review of the pertinent anatomy of joint compartments and main ligaments of the wrist, the authors present the technique of wrist arthrography and CT arthrography. They discuss the normal and pathological patterns. The main indications are the chronic painful wrist and posttraumatic wrist. Imaging can reveal ligament defects (particularly scapholunate ligament, or lunotriquetral ligament), triangular fibrocartilage tears, capsular defects, cartilage thinning, foreign bodies, synovial ganglia or synovitis of the wrist. PMID- 9754338 TI - -Computed tomography arthrography and tendon imaging of the ankle-. AB - Ankle opacification dramatically increases the diagnostic value of CT examination of the foot and ankle. The procedure may be entirely performed on the CT table. The main results and indications of CT-arthrography of the ankle are presented. CT-tenography of the ankle which includes the opacification of a tendon sheath on the CT table, is also described. PMID- 9754339 TI - -Cervical myelography using the lumbar route-. AB - Cervical myelography is obtained after injection of contrast medium into the lumbar subarachnoid space through a lumbar puncture. The patient is prone with a large radiolucent block under the pelvis so that the site of lumbar puncture is at a higher level than that of the cervical spine lordosis. Lumbar contrast is injected gently so that the contrast fluid slides along the ventral aspect of the dural sac and immediately reaches the cervical subarachnoid space. Then AP and lateral (with a horizontal X-ray beam) views are obtained. The patient has to remain firmly immobilized during the whole procedure to avoid blending of the contrast medium with CSF. This procedure is safe and allows cervical myelography to reach the same quality as that obtained through cervical puncture. PMID- 9754340 TI - -How does one question a patient who is suffering still after lumbar disk surgery, before using computed tomography or MRI of the spine?-. AB - Interrogation of patients still painful after lumbar intervertebral disk surgery provides with a series of informations each of them having only an indicative value, while they altogether compound a score helping assess the presence or the absence of a recurring disk herniation. PMID- 9754341 TI - -The diagnosis of localized osteolysis-. AB - The diagnosis of localized osteolysis (synonym: bone cyst, defect) must be analytical. The site, size, and shape of the defect, its limits, assessed according to the Lodwick classification, reflect the activity of the lesion, the type of any periosteal reaction and the presence of a soft tissue mass are analysed successively. Based on these criteria and the clinical and laboratory characteristics, the image can be classified as quiescent or active, allowing guidance of management. PMID- 9754342 TI - -Infarction or chondroma?-. AB - The discovery of an image of central bone calcification raises the differential diagnosis of bone infarction and chondroma. The matrix of chondroma is characteristic of cartilage. It produces typical cartilaginous calcifications: rings, arcs, coarse, irregular grains, moth-eaten appearance. These calcifications are predominantly observed in the centre of the image. They are situated in a bone defect, often multilocular, sometimes accompanied by multiple scratch marks of the cortical endosteum. In contrast, infarction is characterized by the presence of a serpiginous calcified border at the interface between live bone and dead bone. In the centre, the bony trabeculae are still visible on CT sections, in contrast with chondroma. On MRI, chondromas present a heterogeneous lobular appearance on T2-weighted sequences (checkerboard appearance) due to alternating zones of high signal intensity cartilaginous matrix and low signal intensity calcified or ossified fibrous septa. On MRI, bone infarction is characterized by a continuous peripheral line with a marked low signal intensity. PMID- 9754343 TI - -Continuous rotation computed tomography of the bones and joints-. PMID- 9754344 TI - -How to obtain the best results using digitalization-. AB - The main characteristics and the relative interest of the different modalities of medical imaging digitalization are presented as well as their most appropriate clinical indications. PMID- 9754345 TI - -Overexposed film--underexposed film-. AB - Physics of the radiographic exposition are presented, including those of the radiating image, contrast, sensitometry. Based on these principles, the modifications which allow correcting over- and underexposition of the images are explained. PMID- 9754346 TI - -Lateral views of the hip-. AB - Since no true roetgenographic lateral view of the hip is available, several different additional views of the hip have been described to complete the analysis obtained on the AP view. These different additional views of the hip are presented as well as their indications depending on the clinically suspected hip disease. PMID- 9754347 TI - -Arthrography of the hip-. AB - Painful hip with normal X-rays (including false-lateral view of both hips) is the main indication for hip arthrography, searching for focal chondropathy. In the presence of moderate hip degenerative arthritis contrasting with increasing disability, a labrum lesion must be investigated. Such lesions are of two kinds, mainly degenerative (of the superior labrum) or post-traumatic (of the posterior labrum). Arthrography allows the diagnosis of cystic changes located in the acetabulum. It also allows the diagnosis and evaluation of the extent of synovial osteochondromatosis or villonodular synovitis. In three indications, computed tomographies must be added to plain arthrography: posterior labrum lesions, osteochondromatosis (looking for foreign bodies in the acetabular fossa) and cystic changes in the acetabulum (with density measurements). In all cases, a complete conventional radiological assessment must be available. Strict asepsis must be ensured. When osteochondromatosis is suspected, evaluation of articular capacity must be systematic. PMID- 9754348 TI - -Loosening of the complete hip prosthesis. Physiopathology. Therapeutic approach . PMID- 9754349 TI - -Arthrography of the hip prosthesis-. AB - Arthrography of hip arthroplasties is a safe, easy-to-perform procedure. It is mainly indicated when infection of the arthroplasty is suspected: the contrast agent fills bone defects that may be present between normal bone and the prosthesis and reaspiration of injected fluids may lead to bacteriological diagnosis. Another indication for arthrography of hip arthroplasties is unexplained painful hip prosthesis. PMID- 9754350 TI - -Knee prosthesis-. AB - The initial radiological assessment of knee osteoarthritis includes the evaluation of the osteochondral destruction, collateral ligament laxity and the determination of the position of the patella in both the axial and the sagittal planes and the femorotibial angulation in the coronal plane. The different types of knee prostheses are described. Post-operative radiological features of knee arthroplasties are discussed including normal findings and criteria of prosthesis instability and loosening. PMID- 9754351 TI - -Bone tomoscintigraphy in osteoarticular pathology-. AB - Bone single photon emission computed tomography is an effective method for demonstrating partial or total physeal arrest by the proportionality of osteoblastic activity in the physeal regions of interest. The technique complements computed tomography which is not precise in identifying progressive thinning of the physeal bar, and magnetic resonance imaging which does not demonstrate the progressive disappearance of the cartilaginous signal. The radionuclide examination may be a unique imaging method to differentiate between generaled delays in growth and complete fusion at the growth plate. PMID- 9754352 TI - -Lesser known stress fractures-. AB - Stress fractures of the tibia may disclose a longitudinal orientation which is obvious at bone scanning; a mild periostosis may appear on plain films; CT demonstrates a radially-oriented fracture in one aspect of the diaphyseal cortex. A cortical dissection-like vertically oriented insufficiency fracture may involve the medial aspect of the femoral shaft underlying the lesser trochanter; the fracture is concentric to the femoral cortex at CT. Insufficiency fractures of the sacrum may be misdiagnosed on plain films; bone scanning displays a typical H shaped increased uptake which is a specific pattern. Insufficiency fractures of the pubis may appear as tumoral bone destruction; however no soft tissue mass is present at CT which in addition demonstrates normal fat tissue abutting the osseous lesion. PMID- 9754353 TI - -Apatite rheumatism, unusual aspects-. AB - Apatite calcifications may be responsible for unusual aspects. Clinically they may mimick sepsis or soft tissue tumor, and radiologically a bone tumor or soft tissue tumor-like lesion with respect to the erosion of the bone cortex adjacent to an unusual calcification. Diagnosis is based on knowledge of atypical sites of calcification, CT, and mainly, radiological regression of the calcification after a short follow-up. PMID- 9754354 TI - -Morton's neuroma-. PMID- 9754356 TI - [Origin and progression of thyroid epithelial tumors: molecular and cellular mechanisms]. AB - Tumours of the thyroid follicular cell have been intensively studied as a model for investigating the molecular genetics of tumour development in epithelial cells. This review summarises the abnormalities of oncogenes and tumour suppressor genes which have been associated with the major sub-types of thyroid tumour. Although inevitably incomplete, the available data demonstrate well how successive genetic abnormalities drive clonal progression. Comparison of follicular and papillary tumours also reveals a fascinating correlation between genotype and pathological behaviour, consistent with a determining influence of the initiating oncogene on the "route" of tumour development. Finally, the emerging clinical implications are discussed. PMID- 9754357 TI - [The RET gene in thyroid pathology]. AB - The RET proto-oncogene encodes a receptor tyrosine kinase which plays a crucial role during the embryonic development of the enteric nervous system and of the kidney. Cytogenetic analyses of papillary thyroid carcinoma (PTC), a neoplasm which originates from thyrocytes, have revealed that somatic rearrangements of the RET gene are involved in the etiology of a significant proportion of this tumour. Medullary thyroid carcinoma (MTC) which arises from neural-crest derived C-cells is the cardinal disease feature of multiple endocrine neoplasia type 2 (MEN 2), a dominantly inherited cancer syndrome. Recent studies have provided evidence that germline mutations of the RET gene are the underlying genetic events responsible for MEN 2. This review focuses on the role of RET mutations in the pathogenesis of PTC and MTC and summarizes our present knowledge on the consequences of these alterations on the RET tyrosine kinase function. We further describe a transgenic mouse model for hereditary MTC. Mice carrying a MEN 2A allele of RET under the control of the CGRP/calcitonin promoter develop bilateral and multifocal MTC, morphologically and biologically similar to human MTC. PMID- 9754359 TI - [Follicular carcinoma]. AB - Thyroid follicular carcinoma are divided in three subgroups: 1) minimally invasive well differentiated encapsulated follicular carcinoma; 2) invasive well differentiated follicular carcinoma; 3) moderately differentiated follicular carcinoma. Usually diagnosis between encapsulated well differentiated follicular carcinoma and atypical adenoma is difficult. Apart the presence of metastases, histologic criteria to separate these entities are often insufficient. Insofar as all these tumours have a very good prognosis, we think that they could be brought together under the same terminology. Actual morphological differences would be expressed, like in some others localizations, in a histologic grading. This way of classifying appears more consistent with reality and could allow to assume in better conditions, diagnostic uncertainty which exists in this field. PMID- 9754358 TI - [Transgenic mouse models. Their interest in thyroid tumors]. AB - Mouse transgenic models that develop thyroid diseases were generated. All transgenes were driven by the thyroid specific promoter of the thyroglobulin gene. The tissue specificity of the promoter was investigated by using the bacterial chloramphenicol acetyltransferase gene as reporter. The expression of the adenosine A2a receptor resulted in the permanent activation of the cAMP cascade. As a consequence, transgenic mice developed severe hyperthyroidism and a large goiter, demonstrating in vivo the role of the cAMP cascade in the promotion of both function and proliferation of the thyroid cell. These mice constitute a model for autonomous hyperfunctional adenoma and non autoimmune familial hyperthyroidism, where mutant of thyrotropin receptors stimulate the cAMP cascade constitutively. The expression of a mutant of the alpha 1B adrenergic receptor resulted in the constitutive activation of both the cAMP and IP3-CA++ cascades, growth stimulation, hyperfunction, cell degeneracy attributed to the overproduction of free radicals, and development of malignancies. The expression of the SV40 large T antigen promoted the development of aggressive undifferentiated tumors mimicking the phenotype of human anaplastic carcinomas and embryonal tumors. In another transgenic model, the function of the retinoblastoma susceptibility gene product RB1 (and of related proteins) was inhibited by expressing the E7 oncoprotein of human papillomavirus type 16. The result was the development of a differentiated and normofunctional colloid goiter, with progressive development of differentiated malignant lesions. This model suggests the essential role of RB1 and related proteins in the negative control of proliferation that characterizes thyroid cells in the adult. Other transgenic models of thyroid diseases are discussed. PMID- 9754360 TI - [Thyroid papillary carcinoma and its variants]. AB - The authors discuss the histological features of classical thyroid papillary carcinoma, with special emphasis on pathologic indicators related to prognosis. Its different variants are described, including: microcarcinoma, encapsulated variant, pure follicular variant, solid/trabecular variant, diffuse sclerosing and the cytologic variants of tall, columnar and oncocytic cell. PMID- 9754361 TI - [Thyroid insular carcinoma]. AB - This review article illustrates the several histological and immunohistochemical patterns of thyroid insular carcinoma and their associated disease. Differential diagnosis are also discussed. Poorly differentiated thyroid carcinomas are overviewed. The interest of transgenic mice models is presented. PMID- 9754362 TI - [Oxyphilic cell and clear cell carcinoma of the thyroid]. AB - Oxyphilic cell (Hurthle cell) carcinomas of the thyroid gland, variant of follicular carcinoma, are more malignant than follicular non oxyphilic cell carcinomas with a similar size and degree of invasiveness. Gross features, microscopic features of oxyphilic, clear and bicolored cells carcinomas and diagnostic techniques are related with a differential diagnosis with papillary oxyphilic cell carcinoma. PMID- 9754363 TI - [Anaplastic carcinoma of the thyroid]. AB - Anaplastic carcinoma of the thyroid, also called undifferentiated carcinoma, is a rare variety of thyroid carcinoma observed in elderly subjects, usually female. This tumour presents in the form of a cervical mass and is rapidly fatal, usually within several months. Histologically, anaplastic carcinoma consists of a proliferation of spindle and/or giant cells, more rarely squamous cells, rich in cytonuclear abnormalities and mitotic figures with areas of necrosis. Immunohistochemistry confirms the epithelial nature of the proliferation, as tumour cells are generally labelled by low molecular weight cytokeratins; cytokeratin-vimentin coexpression is not rare. Anaplastic carcinoma can occur de novo or secondary to a pre-existing thyroid lesion, particularly well differentiated papillary or follicular carcinoma; in this latter case, study of p53 can be useful to detect the appearance of anaplastic transformation. PMID- 9754364 TI - Primary lymphomas of the thyroid gland: a review with emphasis on diagnostic features. AB - The information about primary lymphomas of the thyroid--a rare form of thyroid tumor and a rare form of lymphoma as well--is dispersed in numerous scientific publications. Recent developments in the field of lymphoid neoplasias, in particular the recognition of mucosa-associated lymphoid tissue (MALT) and the characterization of its distinctive type of lymphomas, has drawn new attention to primary lymphomas of the thyroid and has provided new insights into this field of thyroid pathology. In this paper we review the pathogenesis and the clinicopathologic features of primary lymphomas of the thyroid, we discuss the diagnostic basis and the strategies for differential diagnosis, and we address some recent data that may provide additional pathways to further our understanding of those rare thyroid diseases. PMID- 9754365 TI - [Medullary thyroid carcinoma: evolution of concepts]. AB - This article analyses in a large overview several of the actual problems encountered by pathologist in the management of MTC whatever the diagnostic circumstances. We insist on difficulties upon C cell hyperplasia and early detected disease. Unusual MTC cases are discussed. The need for large multidisciplinary group in studying such tumors is underlined with reference to the French GETC (Groupe d'Etude des Tumeurs a Calcitonine). PMID- 9754366 TI - [Borderline lesions of the thyroid: diagnostic strategy]. AB - Some thyroid neoplasms notoriously cause problems in the histologic diagnosis of their nature (follicular versus papillary for example) or their malignant potential (adenoma versus carcinoma) because of overlapping histologic features. Here, thyroid lesions are presented according to their histologic presentation (vesicular, papillary, trabecular, solid or cystic patterns, fibrosing thyroids and lymphoid infiltrate). For each entity, the most discriminant histologic features are described with emphasis on the diagnostic pitfalls and their histological clues. PMID- 9754367 TI - [Frozen section examination in thyroid pathology. Technique and effective indications]. AB - Frozen section examination for intraoperative diagnosis is often difficult in thyroid pathology. Fine-needle aspiration is widely used in the patients selection for surgery. Such a selection is accompanied by an increase in difficult frozen section cases, whose diagnosis is often differed to permanent sections. Thus, frozen section sensitivity is relatively low. In the era of cost containment, an increasing number of authors suggest that frozen section examination is unnecessary, and that surgical planning could rely on preoperative cytology only. Others consider fine-needle aspiration cytological evaluation and frozen section examination as complementary tools and recommend their association. The authors describe the technical aspects and difficulties of frozen section examination in thyroid pathology, and discuss its interest in surgical planning, in the light of preoperative cytology. Each team has to estimate local thyroid cytology development and accuracy, to define new indications for frozen section examination in thyroid pathology, according to local therapeutic choices. Such an approach could consistently reduce the number of intraoperative consultation for thyroid pathology, without prejudice for the patients. PMID- 9754368 TI - [Fine needle aspiration of the thyroid]. AB - This article precises the technical conditions of fine-needle aspiration, elements of microscopical analyses which establish the diagnostic, complementary technics, specially immunocytochemistry that could enhance the performance. Limits and worrisome histologic changes following fine needle aspiration are also discussed. Fine needle aspiration results are discussed. The text is fully illustrated. PMID- 9754369 TI - [Epithelial tumors with thymus differentiation of the thyroid gland and the neck]. AB - Epithelial tumors with thymus-like differentiation occurring in the thyroid gland and the neck, delineate four entities with distinct histological features and different behaviour. The spindle epithelial tumors with thymus-like differentiation are potentially malignant tumors occurring in the thyroid gland of young patients. They are highly cellular with pattern of spindle cells and tubulopapillary formations and mucinous glands. Carcinomas showing thymus-like differentiation of the thyroid gland are histologically identical to some thymic carcinomas. The ectopic hamartomatous thymoma is strictly benign and occurs in the soft tissues of the lower neck. It is characterized by spindle and solid or cystic islands of epithelial cells associated with adipose cells. The ectopic cervical thymoma is usually benign and histologically similar to mediastinal thymomas. All these tumors are particularly rare and arise from ectopic thymus or remnants of branchial pouch. The differential diagnoses of these tumors can be difficult and include some other proliferations occurring in the neck. PMID- 9754371 TI - Embryonic and fetal development of structures associated with the cerebro-spinal fluid in man and other species. Part I: The ventricular system, meninges and choroid plexuses. AB - Little is known about the development of the central nervous system (CNS) in humans. Ethical considerations preclude experimental studies in this field, and as a result most available data on human ontogenesis are descriptive. Comparative anatomic and embryologic studies have demonstrated that the main developmental milestones are conserved across species, and their results can be used to suggest a likely scenario for human development. The development of the ventricles, meninges, and choroid plexuses are discussed in this article. The central cavity of the neural tube is formed during neurulation, which occurs during the fourth gestational week. The first milestone is occlusion of the spinal neurocele (the central canal in the neural tube) shortly after neurulation. This prevents free communication between the ventricular system and the amniotic cavity. The second milestone is development of the meninges, which separate the central nervous system from the rest of the body. The embryonic origin of the meninges varies across species. In birds (and probably in mammals), the spinal meninges are derived from the somitic mesoderm, the brainstem meninges from the cephalic mesoderm, and the telencephalic meninges from the neural crest. Differentiation of the meninges, which involves formation of the subarachnoid space, occurs early, before the cerebrospinal fluid (CSF) begins to flow around the CNS. During ontogenesis, the meninges play a key role in regulating the growth of underlying nervous structures. They induce the formation of the superficial glial limiting layer and stimulate the growth of precursors located in the superficial blastemas of the cerebellum and hippocampus. The choroid plexuses are complex specialized structures that produce most of the CSF. Their epithelium derives from the neural tube epithelium and their mesenchyma from the meninges. Of the many enzymes produced in the choroid plexuses, some reflect the pivotal metabolic role of these structures (alkaline and acid phosphatases, magnesium-dependent ATPase, glucose-6-phosphatase, thiamine pyrophosphatase, adenylate cyclase, oxidoreductase, esterases, hydrolases, cathepsin D, and glutathion S transferase). The two enzymes that are crucial to the production of CSF are Na+/K+ ATPase and carbonic anhydrase. Inactivation of catecholamines is mediated by catechol-O-methyltransferase and by the monoamine oxidases A and B. The morphology and synthesis profile of the choroid plexuses changes during development, although little is known about these changes in humans. PMID- 9754370 TI - [Hamartomatous adiposity of the thyroid]. AB - Report of one case of hamartomatous adiposity of the thyroid gland. Only eight cases have been reported. The lesion is composed of thyroid tissue and mature adipose elements. Previously reported cases are reviewed and the pathogenesis is discussed. PMID- 9754372 TI - [Arrhythmogenic right ventricular dysplasia and cardiomyopathy. Clinical and anatomic-pathologic aspects, nosologic approach]. AB - Arrhythmogenic right ventricular dysplasia is a polymorphous clinical entity. Its diagnosis is difficult in incomplete forms or at the onset of the disease. The diagnosis is based on the association of clinical, electrocardiographic and electrophysiologic signs which are the result of a specific pathological structure, consisting of fibromuscular bundles isolated from each other by fatty tissue resulting from apoptosis and/or the basic dysplastic phenomenon. These fibers are oriented in a parallel direction and connected at their extremities with normal myocardium. These fibromyocyte bundles seem to constitute a tissue with preferential conduction, which could explain reentry phenomena, and therefore the basis for the pathogenesis of ventricular arrhythmias. The various clinical aspects of ARVD have similar morphological patterns, but a completely different prognosis and outcome. PMID- 9754373 TI - [Pulmonary lymphangioleiomyomatosis. Immunohistochemical and ultrastructural study of two cases]. AB - The authors report two cases of pulmonary lymphangioleiomyomatosis which were diagnosed by surgical biopsies. Immunohistochemical study showed positive staining of the smooth muscle cells using antibodies directed against smooth muscle actin, specific muscle actin, HMB45 and vimentin. Ultrastructural study showed some smooth muscle differentiation features, with numerous myofilaments and some dense bodies near the plasma membrane. In one case, the patient was treated by a double-lung transplantation. LAM is a non-familial disease occurring exclusively in females. The etiology is unknown. This disease can be complicated by chronic respiratory failure. Extrapulmonary sites are not rare, particularly in the uterus. Anti-estrogen treatment can slow the course of the disease. Lung transplantation is actually the only effective treatment at the present time therapy. The differential diagnosis of this disease is discussed. PMID- 9754374 TI - [Value of a computer-assisted screening method (PAPNET) for the detection of infectious cervico-uterine smears]. AB - There have been several reports regarding the accuracy of the PAPNET system applied to the screening for cancerous and precancerous lesions. Based on neuronal networks, this computerized tool was initially trained to select atypical cells. It has been approved in the USA for the re-screening of cervical smears for quality assurance. However, its particular behaviour has not been frequently studied when the system is designed to detect frequent infectious organisms. We report the results of re-screening of 42 inflammatory cervico uterine smears by the PAPNET system. The computerized images were reviewed by two different pathologists, with complete agreement between the two observers in 39 cases. Infectious organisms were detected in only 66% of cases. Trichomonas, mycoses and Gardnerella were diagnosed in 63%, 56% and 87% of cases respectively. No herpetic lesions were identified. The low accuracy of the PAPNET system in the diagnosis of infectious cervico-uterine smears should be taken into account if this system is developed as an exclusive pre-screening method. PMID- 9754375 TI - [Intracranial malignant melanoma. Report of 4 cases]. AB - We report 4 cases of intracranial melanoma without any clinically diagnosed extracerebral location. These tumors presented at different sites and had different histological aspects. Intracytoplasmic melanin was present in all cases, immunohistochemistry confirmed the diagnosis and eliminated a possible glioblastoma or metastatic carcinoma. Clinical and pathologic characteristics of primary pigmented lesions of the CNS are discussed with particular interest in diagnostic difficulties and histogenesis of these lesions. PMID- 9754376 TI - [Cutaneous malacoplakia. An immunohistochemical and ultrastructural case study]. AB - The authors report a case of a 70 year-old man with a past of myelodysplasia and presenting a voluminous lesion of the thigh corresponding to a cutaneous malacoplakia. Histologic study showed a dermo-hypoderma granuloma with numerous Von Hansemann cells containing some Michaelis-Gutmann bodies. Immunohistochemical study showed a positivity of these cells with the antibodies against CD68 (KP1, Mac 387, PGM1), the lysozyme and the alpha-chemotrypsine. Ultrastructural study confirmed the histiocytic origin of this infiltration by showing some regular and voluminous inclusions with a clear center and a peripheral and dense ring, and also some bacteria measuring 3 to 5 microns. Bacteriological study isolated an Escherichia coli. The evolution was favourable after surgical excision and antibiotherapy. Cutaneous malacoplakia is a very rare disease, usually with a perineal localization, and occurring in immunodeficient host. Michaelis-Gutmann bodies are sometimes difficult to identify by light microscopy underlying the rule of the immunohistochemical and the ultrastructural studies to perform the diagnosis. PMID- 9754377 TI - [Lymphoepithelial tumor of the skin]. AB - The authors report an additional case of lympho-epithelial tumor. First described in 1987 by Santa-Cruz et al., this tumor, also called cutaneous lymphadenoma, is rare. The mean age is 44. The lesion is generally located on the face and the clinical diagnosis of basal cell carcinoma is most often evoked. The tumor has a peculiar histological feature with cyst-like cavities composed by an epithelial proliferation and intraepithelial lympho-histiocytic cells. It is considered as a benign tumor. Its origin is still discussed but a pilosebaceous nature seems to be accepted. PMID- 9754378 TI - [Endobronchial granular cell tumor. A report of two cases in the course of staging of ORL cancers]. AB - We report two new observations of endobronchial granular cell tumor discovering by bronchoscopy, during the staging of head and neck cancers. This site remains exceptional; the histologic features are well known. The ultrastructural and especially the immunohistological criteria trend to indicate a neurogenic histogenesis. Although the course is benign in this site and no malignant forms have been reported, local relapses are possible. However, some benign cases may have disturbing endoscopic and histologic features. PMID- 9754380 TI - [Fine-needle cytology and core biopsy of nonpalpable breast lesions. When appropriate?]. AB - The respective roles of fine-needle cytology and core biopsy for the investigation of breast densities and microcalcifications are discussed. For microcalcifications, the type, number, and conditions of development should be taken into account, and the reliability of investigations are directly related to stereotactic visibility, i.e., to advances in digitization. Fine-needle biopsy has been almost completely discarded as a means for investigating microcalcifications but remains useful when performed under ultrasound or stereotactic guidance to investigate densities presumed to be nonmalignant. Doubtful densities are best investigated by core biopsy. PMID- 9754379 TI - [Bilateral scrotal panniculitis in a prepubescent child]. AB - Scrotal panniculitis or scrotal fat necrosis is an uncommon acute scrotal pathology. We report a case of scrotal fat necrosis in a 9 1/2 year-old over weight boy with bilateral tender scrotal masses, located beneath the tests. Pathologic findings were those of subcutaneous fatty indurated masses with lipogranulomatous foci. The etiology of scrotal fat necrosis is unknown. A similar condition has been described in young children exposed to cold. In obese prepubescent boys, a greater sensitivity to cold and a higher saturated fatty acid concentration of the scrotal adipose tissue would induce fat necrosis. Our study of fatty acid composition by gas-liquid chromatography showed an elevation of stearic acid. The spontaneous resolution of scrotal fat necrosis is always the rule and allows symptomatic treatment without surgical investigation. PMID- 9754381 TI - [Indications for diagnosing nonpalpable breast lesions]. AB - The increasingly widespread use of mammographic screening for breast cancer has induced a considerable increase in the number of surgical biopsies. Fine-needle aspiration and microbiopsies can allow to reduce this number. 1) In patients with nodular densities of benign or indeterminate appearance, a negative fine-needle aspiration indicates that no further investigations are needed. In contrast, complete surgical excision is indicated in cases of stellate images. 2) In patients with potentially malignant microcalcifications, fine-needle aspiration is to little value, and microbiopsies should be performed. Indeterminate (type II or III) calcifications are the best indication, since negative microbiopsies may obviate the need for lumpectomy, if the negative predictive value of microbiopsies is sufficiently high in this indication. Focal suspicious microcalcifications (type IV or V) should be removed surgically for both diagnostic and therapeutic purposes. Type IV or V microcalcifications involving a large area of the breast can be investigated by initial microbiopsy; a positive result allows to recommend immediate mastectomy without prior lumpectomy. Fine needle aspiration and microbiopsies should be performed as part of a multidisciplinary diagnostic strategy involving radiologists, surgeons, cytopathologists, and pathologists. This approach is the only means of improving the management of non palpable mammographic lesions and of reducing the number of unnecessary operation. PMID- 9754382 TI - [Fine-needle biopsy and core biopsy in nonpalpable breast lesions. How does one judge with mammography?]. AB - Techniques for localizing and sampling subclinical lesions seen on mammograms are reviewed. Localization of the lesion, the equipment used (fine-needle aspiration or core biopsy), and harvesting techniques are discussed, with special emphasis on quality criteria that should be satisfied to ensure optimal histologic performance. PMID- 9754383 TI - [Fine-needle biopsy under echographic control in nonpalpable breast lesions. Technical aspects]. AB - Fine-needle biopsy under ultrasound guidance is widely used in the diagnosis of subclinical breast lesions seen on mammograms. This minimally invasive procedure requires little time and is reliable for the diagnosis of lesions in a central or peripheral (axillary extension, upper part of the superomedial quadrant, mammary fold) location, or in small-sized breasts. The procedure is done in real time, allowing reliable verification of the specimen. The cytologic results are available almost immediately. Close collaboration between the radiologist and pathologist is essential. PMID- 9754384 TI - [Stereotactic and ultrasound guided fine-needle cytology in nonpalpable breast lesions]. AB - Histologic findings from two consecutive series of fine-needle biopsies of nonpalpable breast lesions were compared. The first series included 243 tumors sampled stereotactically, and the second 198 tumors sampled under ultrasound guidance. The results, together with data from the literature, demonstrated that: 1) the diagnostic efficacy of stereotactic fine-needle cytology of lesions seen on mammograms only as microcalcifications is inadequate; 2) ultrasound-guided fine-needle cytology is an extremely reliable diagnostic tool for lesions that are not seen only as microcalcifications. PMID- 9754385 TI - [Large core of nonpalpable breast lesions]. AB - The development of breast cancer screening has radically changed the diagnostic approach to breast lesions. Stereotactic large-core breast biopsy was developed to reduce the number of unnecessary surgical excisional biopsies in the increasing number of patients with doubtful or suspicious mammogram findings. The methods used to evaluate this new technique are discussed, as well as results in terms of efficacy, factors that may influence efficacy, and difficulties in interpreting large-core biopsies. Recommendations regarding the harvesting and interpretation of stereotactic large-core biopsies are made. PMID- 9754386 TI - [Diagnostic difficulties and limits in imaging]. AB - Fine-needle cytology and core biopsy of the breast done under mammographic or ultrasonographic guidance can assist in decision-making (monitoring, diagnostic or therapeutic procedures). Cytologic studies and histologic evaluation of core biopsies have their own limitations: the samples are small in size, and the histopathological features of the lesions are often complex. Each step of an image-guided diagnostic procedure contributes to the reliability of the method. The characteristics of the mammogram and/or ultrasonogram image provide initial orientation. Whether they are concordant with the results of the cytologic or histologic study is an important factor. The main limitations of stereotactically guided biopsies are ballistic with targets that are difficult to circumscribe (fine microcalcifications or ill-defined density). With ultrasonography, verification of the sampling site in the target is essential. The performances of these procedures are closely dependent on the experience of the operators and on the quality of the cooperation between clinicians, radiologists and pathologists. PMID- 9754387 TI - [Breast cytopathology. Diagnostic difficulties and limits]. AB - Diagnostic difficulties can arise during the cytologic diagnosis of almost all types of breast lesions. "True" difficulties, which are discussed herein, should be differentiated from difficulties due to faulty technique. The frequency of diagnostic difficulties varies across lesions: the proportion of "suspicious" specimens is 4% in adenofibroma, 5 to 7% in cancer, and 17% in epithelial duct hyperplasia, and the false-negative rate in cancer is 3 to 5%. Many difficulties can be overcome by a good knowledge of breast cytopathology. Others are insuperable and should remain so to avoid diagnostic mistakes. In these cases, which should be considered "suspicious", the clearly written documented report should request a histological study. The distinction between duct carcinoma and lobular carcinoma remains difficult, and that between invasive carcinoma and intraductal carcinoma requires a histologic study. PMID- 9754388 TI - [Diagnostic difficulties and limits in breast histopathology in core biopsies (breast microbiopsies)]. AB - Core biopsy is increasingly used for the diagnosis and management of breast lesions. As a result, pathologists are being confronted with new difficulties pertaining to (1) the histological diagnosis on micro specimens (2) the correlation between core biopsy histological findings and mammographic findings, (3) the diagnostic significance of atypical hyperplasia on core biopsies, (4) the impact of tissue displacement on surgical specimens after needling procedures, and (5) the disagreement that can appear between the diagnostic on surgical specimen and on core biopsy. PMID- 9754389 TI - [Mitochondrial DNA and Parkinson disease. Methodologic review]. AB - Parkinson's disease (PD), a disorder of unknown etiology, is associated with the degeneration of dopaminergic neurons in nigro-striatal pathways. MPTP, a meperidine analog, causes parkinsonism in human and nonhuman primates. MPP+, the active metabolite of MPTP, inhibits the activity of respiratory chain complex I. In patients with PD, a reduced complex I activity was found in substantia nigra, skeletal muscle, and platelets. Because complex I is partially encoded by the mitochondrial genome, several studies have searched for mitochondrial (mt) DNA abnormalities in patients with PD. Our aim was to answer the following questions: (1) are there some abnormalities of mtDNA in PD? (2) if there are some, what are these abnormalities? and (3) what is the pathogenic role of these abnormalities? METHODS: The literature review was performed using Medline [National Library of Medicine, Washington] and Current Contents [Institute for Scientific Information, Philadelphia] databases. Periods screened were 1966-March, 1998 (Medline) and March 17, 1997-March 9, 1998 (Current Contents). Keywords were: "Parkinson" or "Parkinson's", and "mitochondrial DNA" or "mtDNA". We limited our research to articles in English and French. RESULTS: Medline search provided 59 articles. Current Contents search provided 22 articles. Twelve articles were found in both databases. Thirty-eight of the 69 articles were either reviews about mitochondrial diseases (19 articles) or original articles not related to mtDNA (19 articles). Our final selection included the remaining 31 articles. PMID- 9754390 TI - Making a difference. PMID- 9754391 TI - MedBytes. PMID- 9754392 TI - VitalStats. PMID- 9754393 TI - Coming in first. PMID- 9754394 TI - Show us the money. PMID- 9754395 TI - The gene finders. PMID- 9754396 TI - Drugs detected in patients suspected of acute intoxication. AB - Drug screens were performed for 434 adult patients who presented to the Parkland Memorial Hospital Emergency Department with suspected acute drug overdose. The screening consisted of analysis of urine by automated high performance liquid chromatography (REMEDi) in combination with qualitative EMIT immunoassays. Selected patients also had ethanol measured in blood, salicylate and acetaminophen measured in serum, and urine specimens analyzed qualitatively for cannabinoids. Most patients (83.4%), regardless of age, race, or gender, had evidence of consumption of at least one drug. The drugs detected most often were ethanol (30.0%) and cocaine (23.7%). At least one of the nine most common drugs of-abuse (amphetamines, barbiturates, benzodiazepines, cannabinoids, cocaine, ethanol, opiates, opioids, and phencyclidine) was detected in 64.5% of the specimens, and combinations of these drugs were present in 45.4%. For most drugs, age, gender, ethnicity, time of day, day of week, and indication for screening could not be used to predict the drug screen result. PMID- 9754397 TI - Beyond tort reform. AB - Organized medicine has spent a great deal of time, energy, and money attempting to revise the legal tort system. Yet, change, if any, has been incredibly slow. There are many reasons for this. Tort law has been a part of American jurisprudence for hundreds of years. In addition, most state legislatures are populated with large numbers of attorneys. This paper explores the economic factors that underlie the litigation process in medical negligence/malpractice cases. It suggests that the current tort system is not as antimedicine as physicians commonly believe; rather, it is physician-friendly. Presented here is a more efficient and cost-effective method of addressing medical negligence/malpractice cases. An exclusive relationship between the liability insurance carrier and a defense law firm is proposed. Rather than using the old billable hours system to charge for its services, the defense law firm negotiates a yearly retainer based upon a percentage of the annual liability insurance premiums paid. How this relationship would result in a more efficient and cost effective approach to the present tort system is examined. PMID- 9754398 TI - [Gamma-oryzanol: an important component in rice brain oil]. AB - Gamma-oryzanol, a mixture of ferulic acid esters of sterol and triterpene alcohols, it occurs in rice bran oil at a level of 1 to 2%, where it serves as natural antioxidant. Recent research has shown that gamma-Oryzanol can lower the cholesterol levels in the blood, lowering the risk of coronary heart disease, besides that, also has been used in Japan like natural antioxidant in foods, beverages and cosmetics. This review refers to aspects about gamma-Oryzanol, like its physiochemical properties, its presence in the rice bran oil, its antioxidant and hypocholesterolemic activity, as well as, identification, quantitation and extraction methods. PMID- 9754399 TI - [Weight-for-height in adults: comparison of classifications adjusted and non adjusted by frame size]. AB - The present study is to analyze the concordance, agreements and divergence of anthropometry nutritional classification of weight-height (WH) in adults, using criteria that include frame size adjustments and no adjustment at all. 224 adults were studied (127 female and 117 males) from the "Simon Bolivar" University Administrative Employees Health Project, 1993. Using as basis, the variables weight, height, wrist circumference and elbow breadth, we determined: a) frame size by wrist circumference methods (WC) (Grant, 1980) and elbow breadth (EB) (Frame index 2 by Frisancho, 1989); b) classification by weight-height (WH) according to table by frame size (Frisancho, 1984). 57%, 38% and 6% corresponded to small, medium and large frame sizes, by WC. 16%, 60% and 25% by EB. When classifying by WH those results showed differences between 16-25% in female and 15-21% in males. When contrasting the three criteria, it was observed a bigger coincidence between WH without frame size adjustment and WH adjustment by EB. The smallest coincidence between WH adjusted by WC method and weight height without frame size adjustment in the whole group, while male and female got the biggest coincidence in WH adjustment by WC and HW without frame size adjustment. The smallest concordance (k = 0.37) was obtained when contrasting WH frame size adjustment by EB vs weight-height without adjustment in female, and biggest concordance (k = 0.60) when contrasting WH by WC and without adjustment in males. This results show that, there are significative differences in nutritional classification of weight-height in adults adjusted and non adjusted by frame size within the same group of persons. PMID- 9754400 TI - [Nutritional status of full-time students at public schools. Sao Paulo, Brazil]. AB - The nutritional status and some of their conditioning factors of 257 students from 7 to 13 years of age registered at the Integrated Public Education Centers- CIEP's in American, state of Sao Paulo, are analyzed. Height of students is used to determine the nutritional status, with the classification based on standard deviation units (height/age Z score). A 15.6% ratio of, children with chronic malnutrition (HAZ < -2) is observed. Statistical analysis shows that the child's height for age Z score is influenced by per captia family income and attendance today-care centers during preschool age, among other variables. That shows the importance of life conditions prior to the child's entry to CIEP and which have certainly affected his growth throughout periods when a child is biologically most vulnerable. PMID- 9754401 TI - [Effect of altitude on iron absorption]. AB - Iron bioavailability was evaluated in people living in high altitudes. Absorption was estimated from a reference dose of ferrous ascorbate and from a standard diet of wheat flour, using extrinsic tag radioisotope technique of 55Fe and 59Fe. Twenty four volunteers, healthy women, with ages ranging from 28 to 45 years, participated. Of those, eleven lived at 3450 meters above sea level (m.a.s.l.) in Huancayo city-Peru (study group), and 13 lived in Santiago de Chile at 630 m.a.s.l. (control group). Iron absorption from reference dose of ferrous ascorbate was 32.0% and 31.1% in the study and control groups respectively. The geometric mean of iron absorption from the standard diet, corrected to 40% of absorption of reference dose, was 9.0% and 6.9% in the study and control groups respectively (NS). The results suggest that altitude does not produce a high iron absorption in highlander residents. PMID- 9754402 TI - [Diet as a determinant of central nervous system development: role of essential fatty acids]. AB - Nervous system is second to adipose tissue in containing the highest lipid concentration. Membrane phospholipids possess a high content of long chain polyunsaturated fatty acids (LCPUFA) of the n-3 and n-6 families, derived from the corresponding essential fatty acids. Docosahexaenoic acid (22:6n-3) is found in the highest concentrations in the gray matter and the photoreceptors of the retina. n-3 LCPUFA deficiency in infants, mainly if born preterm, modifies neural functions causing learning disabilities and visual function abnormalities. The adequate lipid nutrition of the mother before and during pregnancy and in breast feeding determine the lipid transfer of fatty acids to the fetus and infant, respectively. LCPUFA are conditionally essential in preterm infants, born with lower lipid depots. The formulation of infant formulas, mainly for preterm babies, should include adequate proportions of n-3 and n-6 LCPUFA. PMID- 9754403 TI - Methionine supplementation of soya products: effects on nitrogen balance parameters. AB - Soybean protein is one of the best quality foods available. Contribution of soy to human nutrition increases because of its overall positive nutritional profile, low cost, high protein and excellent functional properties. Addition of methionine to rat soybean diets improve biological value of soy protein. Few studies on methionine fortification of soya protein were carried in infants, but fortification of baby formulas with this amino acid is usually found. This study was carried out to demonstrate in malnourished children that the effect of methionine supplementation of soya milk and soy isolated protein, as well as to compare with their results to cows' milk. A total of 30 malnourished children, 1 to 3 years old, admitted to our metabolic unit and distributed in groups of 6 children were studied. They were fed experimental formulas with cows' milk, soya milk, soya milk plus methionine, soya isolated and soya isolated plus methionine. Nutrient compositions of formulas were calculated to be similar to mothers' milk. DL-methionine, 1.5 g per 100 g protein content was added to soya milk and soya isolated formulas. Two nitrogen balances, 3 days each, were carried out. Fecal and urinary nitrogen, serum proteins, creatinine and urea in serum and urine were followed during the study. Results showed differences of intake and retention of nitrogen between some of the groups, but there were no statistically significant differences on protein absorption in the groups. No differences were demonstrated in serum proteins, total nitrogen and other serum and urine parameters analyzed. Cows' milk fed children presented the highest nitrogen retention in both balance studies. The addition of methionine to the soya milk formula increased the nitrogen retention, not reaching the cows' milk levels and did not have the same effect when added to the isolate soy protein. PMID- 9754404 TI - Comparative study of beans from vine and bush type of vegetative growth. Effect of storage on cell wall components as factors in increasing cooking time. AB - Four samples each of black beans representing two types of vegetative growth were collected from farmers' fields in four locations in Guatemala. Soon after collection, samples were stored at 4 degrees and 38 degrees C at ambient relative humidity and subsamples were withdrawn at 0, 45, 90 and 135 days of storage for determination of water absorption, cooking time and analysis of neutral- and acid detergent fiber, cellulose, hemicellulose and lignin. The fiber fraction analysis were done on samples of 0, 45 and 90 days of storage. Water absorption for all 4 samples of the bush type was similar at both storage T, however the samples stored at 38 degrees C and at 135 days absorbed more water than when stored at 4 degrees C. The 4 vine types of beans showed different water absorption rates, with two showing patterns similar to those beans of the bush type and two which did absorbed water at a very slow rate. For both types of beans stored at 4 degrees C, cooking time decreased from 0 to 135 days of storage. On the other hand for all bean samples of the two types cooking time increased when stored at 38 degrees C. Analysis of variance showed highly significant effects due to plant type, days of storage, temperature and locality, and for some interactions. Analysis of variance of the fiber fractions showed high significant differences for days of storage for NDF, ADF, cellulose, hemicellulose, and lignin. Plant type gave significant differences for cellulose and hemicellulose. Highly significant differences for hemicellulose were found for the interactions of type x days, type x temperature, locality x type, and type x days x temperature. The rate of synthesis of the 5 fractions were calculated by simple regression analysis. For the bush type of beans some synthesis occurred at 4 degrees C, but it was enhanced when stored at 38 degrees C. For vine type of beans at 4 degrees C relative high rates of synthesis were observed, which were higher at 38 degrees C for NDF, hemicellulose and lignin. Cooking time and fiber fraction contents were subjected to regression analysis. The correlations at 38 degrees C were higher than at 4 degrees C for all fractions for both types of beans, but statistical significance was obtained only for NDF, ADF and cellulose for vine type of beans. These data show therefore that synthesis of cell wall structure fractions, and not only lignin formation, are responsible for the increase in cooking time observed upon storage at high temperature. PMID- 9754405 TI - [Effect of cooking on protein digestibility of sorghum (Sorghum bicolor (L.) Moench)]. AB - The changes in protein digestibility that occur during cooking have interested many scientists. In this study the effect of cooking sorghum in water on the in vitro protein digestibility (IVPD) was evaluated using sorghum grains not containing tannin (SST), and grains containing detoxified tannin (SPD). The results were compared with rice and maize. The effect of sulfites present in the water used for cooking was also determined. The IVPD of sorghum excent of tannins before cooking was 71.1% smaller (p < 0.05) than that obtained for com (80.8%), polished rice (90.6%) or sorgum with detoxified tannins (80.4%). After cooking in water the IVPD decreased to 23.1%, 66.3%, 3.1% and 3.2% for SST, SPD, polished rice and corn endosperms, respectively. The IVPD of SST and SPD treated with 0.1M sodium bisulfite was 65.2 and 50.1%, which corresponds to a decrease in IVPD of 8.0 and 37.7%, respectively. Similar results were obtained when 0.1M sodium metabisulfite is added to the cooking media. These findings demonstrate that sulfites inhibit the sudden decrease of the IVPD of cooked sorghum grains, and suggest that these compounds may block the formation of disulfide bridges (-S-S-) among the gamma-kafirins molecules located on the surface of the sorghum protein bodies or possibly other factors involved which will be later studied. PMID- 9754406 TI - [Lipoxygenase and trypsin inhibitor inactivation in soymilk processing by direct milling and ultra high temperature (DM-UHT)]. AB - Soymilk production by a new process based on Direct Milling of soy grits and Ultra High Temperature (DM-UHT) has been studied at laboratory scale in order to evaluate solids and protein extraction, lipoxygenases (LO) and Trypsin Inhibitors (TI) inactivation during milling and heat-treatment steps. For TI measurements in soy extracts and soymilk a modification of the classical Kakade method (17) was used. Highest extraction yields were accomplished at 70 degrees C and 2 minutes milling of soy grits. LO was appreciably inactivated when using, as dispersing medium for milling, 0.01M sodium carbonate (Residual Activity 14%) instead of water (Residual Activity 46%), so in this way lower levels of undesired substances can be generated. LO destruction in the resulting suspension was finished by a short heating (30 seconds) from 70 degrees C to boiling temperature (96 degrees C). On the other hand, TI were not fully inactivated in milling nor even in the steaming step at 96 degrees C for many minutes. The TI were inactivated to the accepted levels for soymilk in the final UHT step at 135 degrees C and 2 minutes, being possible at the same time to carry out the simultaneous microorganisms destruction. PMID- 9754407 TI - [Methods for extracting chitin from shrimp shell waste]. AB - Shrimp shell waste obtained from several industrial freezing-purchasing plants of Guaymas, Sonora, Mex., was studied as a source of value-added chitin biopolymers. In part I, the effect of different isolation conditions on the chitin yield and chemical characteristic, was investigated. Protein and mineral matter were removed with alkali and acid treatment respectively. A 2x2x3 factorial a way of a completely randomized design was used in order to evaluate the effect of the process variables, namely, NaOH concentration (0.4 and 2%) during the deproteinization and HCl concentration (3 and 5%) carried out at 40, 50 and 60 degrees C. The best processing conditions were desproteinization with 2% NaOH, and demineralization with 5% HCl at 50 degrees C, in terms of final ash and chitin content and yield. In part II, a selection of methods of isolation of chitin and chitosan was studied in order to establish the best conditions for scaling up a process to pilot plant level. The processing conditions were selected from reported methods as well as from those defined in part I. Purity of chitin samples was determined in terms of residual protein, ash and chitin each one to produce high quality chitin (0.00% protein, 0.01% ash, 99.99% chitin) and standard grade chitin (0.00% protein, 0.09% ash, 99.13% chitin). Both products were considered as of adequate quality and their manufacture process could be scaled up by further optimization of the processing conditions. PMID- 9754408 TI - A test to detect cane-sugar-honey. AB - A manual procedure for simple detection of cane sugar honey is described in terms of a visual reaction of honey, water and diethyl ether mixtures. Analyses of 30 samples of genuine honey were contrasted with 30 samples of cane-sugar-honey from Venezuela, and discriminated correctly by the proposed method. PMID- 9754409 TI - [Evaluation of iron contents in foods of the basic diet]. AB - The components of the basic food diet available at the food markets in Curitiba were evaluated according to their contents of iron. The method used for this analysis was atomic absorption spectrophotometry. The results obtained for samples of wheat flour were (mg/kg): 9.40 to 17.60; bread 7.04 to 12.80; coffee 45.76 to 49.21; potato 3.56 to 5.65; tomato 6.94 to 15.10; banana 7.59 to 15.20; rice 5.49 to 11.06; bean 59.84 to 78.75; meat 9.48 to 34.29; pasteurized milk 0.33 to 1.20; soy-bean oil 0.47 to 0.76; refined sugar 1.02 to 1.53 and margarine 1.50 to 2.06. There is no specific legislation in Brazil defining an upper limit of tolerance of iron in the food, except for soy-bean oil and margarine with a maximum of 1.50 mg/kg. The levels of iron analysed here were within the scale of variation reported by several authors from other countries. PMID- 9754410 TI - [Presence of total coliforms, Escherichia coli and Listeria sp. in enteral formulas]. AB - The presence of total coliforms, Escherichia coli and Listeria sp. was evaluated in 65 samples of enteral nutrition formulas. In more than the 75% of the samples made up from cooked vegetables, fruits or meat broth, the score level of total coliforms was of 10(4) UFC/g. In 12-31% of the different enteral food formula, E. coli was isolated in levels ranging form 3.0 x 10(2) to 2.1 x 10(4) UFC/g. Enteral nutrition formulas made out of fruits and those elaborated with meal broth presented this agent more frequently and in bigger quantities. Listeria sp. was isolated in 17% of the fruit preparations and in enteral formulas made with milk. L. grayi, L welshimeri and L. innocua were the species found. PMID- 9754411 TI - [Lipid composition of eggs from hens fed with marine protein and fat products]. AB - This study describes the composition of shell eggs produced by hens fed marine sources of feeds and eggs from hens fed only vegetable ingredients, as a control group. The nutritional characteristics of eggs, yolk fatty acids profile and their cholesterol, triglyceride and phospholipid contents, were determined. Hens fed marine feeds produced eggs with significantly less cholesterol and more triglycerides and phospholipids, than those produced by hens fed only vegetable feeds. Even though there were no significant differences between saturated, mono and polyunsaturated fatty acids, marine products significantly increased n-3 fatty acids (7.13 +/- 0.83 in marine fed hen's eggs vs. 1.77 +/- 0.55 g/100 g in vegetable fed hen's eggs) and n-6 fatty acids (15.71 +/- 2 in marine fed hen's eggs vs. 20.88 +/- 2.32 g/100 in all vegetable fed hen's eggs). The eggs produced from hens fed marine products compares favorably with eggs known as "enriched" can have an important place in the diet especially for high risk population groups. PMID- 9754412 TI - [Nutritive value and acceptability of chinchilla's meat]. AB - (Eriomys lanigera y brevicaudata) Food value and acceptability of chinchilla's meat with different thermic treatments were studied. Chinchilla's in captivity of the Province of Jujuy were used. They were divided into three groups: raw and treated with dry and wet heat and the following analysis were carried out: moisture, protein, ether extract, ashes, iron (A.O.A.C.), cholesterol (enzymatic method), Net Protein Ratio (NPR), Digestibility and acceptability tests were also done. Moisture content of raw chinchilla's meat was 68.24 g/100 g and the meat treated with wet and dry heat had moisture of 65.09 g and 62.49 g/100 g respectively. Protein percentage of raw chinchilla's meat was 20.03 g/100 g, no significant differences were observed in the meat thermally treated. The same tendency was observed concerning the ether extract content (11.26; 12.00 and 12.92 g/100 g for raw meat and the one treated with wet and dry heat respectively). No difference was observed among the three types of meat as far as the ash content is concerned. The mg of iron in raw meat were higher (13.76 mg/100 g) than in thermally treated meat (12.43 and 12.37 mg/100 g for wet and dry heated respectively). NPR and digestibility of thermally treated meat were 5.26 and 5.65; 96.75 and 96.84 (for wet and dry heated meat respectively); both were similar to and higher than casein. The acceptability of chinchilla's meat treated with wet heat (94%) was higher than that treated with dry heat (90%). It can be concluded that chinchilla's meat thermally treated has both good food value and acceptability. PMID- 9754413 TI - Clinical and EEG analysis of mesial and lateral temporal lobe seizures. AB - The analysis of the temporal lobe seizures through video-EEG systems shows that they often consist of a sequence of clinical and EEG features which may suggest the localization and the lateralization of the epileptogenic lobe. We analyzed clinical and EEG features of 50 temporal lobe seizures which were separated in group 1 with 25 patients (related to mesial temporal sclerosis) and group 2 with 25 patients (other neocortical temporal lesions). Among the auras, the epigastric type was the most frequent and predominated in group 1. There were differences between the two groups, considering dystonic and tonic posturing and versive head and eye movements. Dystonic posturing was always contralateral to the ictal onset and was considered the most useful lateralizing clinical feature. Ictal speech, spitting and blinking automatisms, prolonged disorientation for place and a greatest percentage of postictal language preservation occurred in right temporal seizures. Postictal aphasia and global disorientation predominated in left temporal seizures. EEG was important for lateralizing the epileptogenic lobe, specially considering rhythmic ictal activity and postictal findings. PMID- 9754414 TI - Background and paroxystic activities on AIDS patients' EEG. Relation with urea and creatinine seric concentration. AB - The HIV is responsible for important metabolic and structural alterations of the brain. This affected brain must react to continuous systemic metabolic fluctuations. We search for possibly resulting cerebral electric disturbance that could be found by EEG exploration. Sixty-three AIDS patients ranked as CDC group IV had their EEG background rhythm measured, and were appointed to mutually exclusiding groups delimited by medians' values of urea (24 mg/dl) and creatinine (0.9 mg/dl) seric concentrations. These groups were independently formed for each of the parameters utilized, and each data pair generated therefrom were compared between themselves to verify whether there were differences in background rhythm and the occurrence of paroxysmal activity. Background rhythm and paroxysmal activities have not statistically differed between the group whose creatinine values were lower than 0.9 mg/dl and the group whose creatinine values were equal or higher than 0.9 mg/dl. Background rhythm has not statistically differed between the group whose urea values were < 24 mg/dl and the group whose urea values were = 24 mg/dl; contrariwise, the occurrence of paroxysmal activities in these groups has significatively differed, being higher in the patient group whose otherwise normal urea values exceeded 24 mg/dl (p = 0.02). PMID- 9754415 TI - Sporadic Creutzfeldt-Jakob disease. A clinico-neuropathological analysis of nine definite cases. AB - The authors have analyzed clinico-neuropathologically nine cases of the definite sporadic form of Creutzfeldt-Jakob disease (CJD). All cases were female, with mean age of 62.7 years. Eighty-nine percent of the patients exhibited prodromal and initial psychiatric symptoms; definite signs of dementia, and myoclonus were present in 100% of cases. The EEG was abnormal in all cases and pseudoperiodic paroxysms were present in 56% of the patients. Their evolution time ranged from 3 to 19 months. Neuropathologically, brain and cerebellar atrophy, spongiosis, astrocytosis and neuronal loss were present in 100% of the patients. In 5 (56%) of these 9 cases, prion protein (PrP) amyloid plaques were detected in the cerebellum, by optical- and electronmicroscopy. There was a positive correlation between the number of plaques and the evolution time. The authors outline the similarities of their cases in the elderly with the new variant of CJD described in young people. PMID- 9754416 TI - Preliminary adaptation into Portuguese of a standardised picture set for the use in research and neuropsychological assessment. AB - Pictorial stimuli and words have been widely used to evaluate mnemonic processes in clinical settings, neuropsychological investigations, as well as in studies on the mechanisms underlying the phenomena of memory. However, there seem to be few studies of standardisation of pictures for research in this field. The present paper aimed at adapting the use of a set of pictures standardised for English speaking subjects for Portuguese speakers. Name agreement of 150 pictures was assessed in 100 high-school students. Ninety pictures were found to present the same name for over 90 subjects. Results yield data that may help create more controlled tests for the study of memory for pictorial stimuli in Brazil. PMID- 9754417 TI - Normal cerebrospinal fluid values in full-term gestation and premature neonates. AB - Results of cerebrospinal fluid (CSF) examinations from 77 high-risk neonates were reviewed. The mean CSF white cells (WBC) count was 4.5 cell/mm3, being two standard deviations above the mean 11.7 cells/mm3 in the full-term gestation neonate group; in the premature neonate one, the mean CSF WBC count was 5.1 cells/mm3, being two standard deviations above the mean 16.7 cell/mm3. PMNs (polymorphonuclear leukocytes) were present in less than 40% of those children, being the mean PMN percentage 4.2% and 0.6%, the mean ANC (absolute neutrophil count) was 0.3/mm3 and 0.06/mm3, in full-term gestation neonate group and premature neonate one, respectively. The mean CSF protein concentration is significantly greater in those premature neonates (101.2 mg/dl) compared with that in term neonates (77.6 mg/dl). The average glucose was just the same in both groups (67 mg/dl). All of these values were from patients who underwent nontraumatic cisternal puncture, with no red blood cells (RBC/mm3 = 0). Traumatic puncture, even up to 500 RBC/mm3, interfered on CSF parameters. PMID- 9754418 TI - Histochemical study of the skeletal muscle in children with chronic renal failure in dialysis treatment. AB - Among the modifications occurring in the uremic organism, in addition to the consequences of dialysis, myopathy and peripheral neuropathy are very significant. Children are particularly affected, as their growth and development are jeopardized. Histochemistry of muscular biopsy was used to study eighteen children with end-stage renal failure under dialysis during a ten-month period. According to our results, the skeletal muscular tissue presented the following types of alterations: atrophy, type grouping, lipidosis, glycogen depletion and mitochondrial proliferation. PMID- 9754419 TI - [Immunocytochemical analysis of the inflammatory infiltrate in inclusion body myositis and other neuromuscular disorders with rimmed vacuoles]. AB - Among 1400 muscle biopsies, we found 16 cases with rimmed vacuoles whose diagnosis were sporadic inclusion body myositis (IBM) (4 cases), juvenile spinal muscular atrophy (6 cases), distal myopathies (3 cases), limb-girdle muscular dystrophy (2 cases), and peripheral neuropathy (1 case). Monoclonal antibodies reactive for T lymphocytes and subsets, B lymphocytes, macrophages, natural killer cells, immunoglobulins, and complement were used to analyze the inflammatory infiltrate. The analysis was quantitative and according to the site of accumulation (interstitial, endomysial, and perivascular). The immunocytochemical analysis showed CD8+ lymphocytes in the interstitial in most cases, occasionally inside of muscle fibers, and rarely in the perivascular region. The IBM cases had an increased number of CD8+ lymphocytes comparing with the other diseases. CD8+/CD4+ ratio was increased in IBM compared with the other diseases. Macrophages were frequent in IBM, distal myopathy, and one case of limb girdle muscular dystrophy. Natural killer cells were frequent at interstitial. PMID- 9754420 TI - [Intrathecal opioids in chronic non-malignant pain: relief and life quality]. AB - The use of opioids for treatment of non-malignant pain is controversial. The evaluation of pain relief and of the quality of life of 11 severely incapacitated chronic non-cancer pain patients treated with long term intrathecal infusion of opioids trought implantable pumps was performed. The mean duration of pain complaints was 5.3 years. The mean pain intensity was 8.6. In 7 patients, pain episodes lasted at least 6 hours daily. The mean duration of the therapy was 19.6 months. After the treatment the mean pain score became 3.9. In only 1 patient, the duration of pain episodes was still longer than 6 hours. Quality of life improved in 36.36% of the cases. The long term spinal opioids through implantable pumps for non-malignant pains results in pain relief but not necessarily improves the quality of life. PMID- 9754421 TI - [Systemic arterial hypertension and psychiatric morbidity in the outpatient care setting of a tertiary hospital]. AB - Arterial hypertension is one of the most important risk factor for cardiovascular disease, the main cause of death in Brazil. Hypertensive patients that have treated in tertiary care hospitals have shown elevated co-morbidity including psychiatric disturbances. Our objective is to study psychiatric co-morbidity among severe hypertensive patients. This study was performed in an out-patient clinic of tertiary medical care setting. Forty-one patients were enrolled in this research (26 women, 15 men). They were submitted to a clinical interview and answering the PRIME-MD, a specific questionnaire for diagnosis of psychiatric disturbances (by a general practitioner). Frequencies of psychiatric disturbances were different in men and women: 63.4% of the women in this study showed some type of psychiatric disturbance versus 36.6% of men (p = 0.012). The majority of the diagnosis were mood disturbances, mainly depression associated or not with anxious disturbances. Mean age of psychiatric disturbance patients was 47.1 years versus 59.3 years in the patients without psychiatric disturbances (p = 0.0049), showing the presence of psychiatric disturbances in younger patients. Other factors as systolic arterial blood pressure, diastolic arterial blood pressure and body mass index did not show any differences associated with psychiatric disturbance. We conclude that there is a great co-morbidity between high complexity hospitals hypertensive patients and that this type of disturbance is more frequent in women and in younger patients. PMID- 9754422 TI - [Memory complaints and the diagnosis of dementia]. AB - Ageing is often associated with the decline of some cognitive abilities, although in most cases these losses do not progress to full blown dementia. The current study aimed to evaluate the association between subjective memory complaint and the diagnosis of dementia among the elderly assessed at the Old Age Unit of the Department of Mental Health of Santa Casa de Sao Paulo-Brazil between February and December 1997. Subjects were assessed with the SRQ-20 and MMSE. Further clinical information was also collected to allow for the diagnosis of mental disorders according to ICD-10 diagnostic guidelines. Fifty-nine percent of the 220 patients included in the study complained of memory difficulties. Seventy-one percent of the complainers were women, although there was no clear statistical association between sex, education, marital status and living conditions with the memory complaint. There was a trend for memory complainers to present with higher scores on the SRQ-20(p = 0.122). The complaint of memory difficulties had a sensitivity of 76.2%, specificity of 47.8%, positive predictive value of 36.9%, and negative predictive value of 83.3% for the diagnosis of dementia. Memory complaints are frequent among the elderly, particularly among those with more severe depressive and anxiety symptoms. The subjective experience of memory difficulties has a low predictive value for the diagnosis of dementia. The identification of "at risk" cases should, instead, be based on new neuroimaging and genetic methods. PMID- 9754423 TI - [Visual and volumetric analysis with magnetic resonance of hippocampus formations in a group of patients with clinical diagnosis of temporal lobe epilepsy]. AB - The purpose of this study is to test the sensitivity of the volumetric analysis compared to the visual analysis of the hippocampal formations of a group of 153 patients with clinical diagnosis of temporal lobe epilepsy and candidates to temporal lobectomy, evaluated by magnetic resonance (MR), using a 0.5 Tesla machine and inversion-recovery T1-weighted images and 5 mm coronal slices. There was agreement between the prospective visual analysis and another retrospective visual analysis carried out by two independent observers (C = 0.748 and C = 0.720). There was also agreement between the retrospective analysis of the two investigators (C = 0.733). There was genuine agreement (C = 0.788) between the results of the qualitative and quantitative analyses carried out prospectively. Volume measurements showed a nonsignificant tendency to lateralize more cases of clinically presumed hippocampal atrophy. Our results confirm the reliability of a qualitative visual analysis and indicate the utility of hippocampal volumetry as a supplementary, objective and quantitative measure of hippocampal sclerosis. PMID- 9754425 TI - [Diagnosis of metabolic diseases of the nervous system in children through ultrastructural analysis of non cerebral tissue]. AB - Although biochemical and molecular genetic analysis are the most precise methods for the diagnosis of metabolic diseases, morphological studies remain a very important diagnostic method mainly in countries like Brazil, where clinical laboratories are unable to perform most of the exams required for the diagnosis of these diseases. Moreover, pathologic evaluation is the single diagnostic method for some disorders whose metabolic defect is unknown such as neuronal ceroid-lipofuscinosis, infantile neuroaxonal dystrophy or Lafora disease. We present our experience with ultrastructural analysis in 582 exams of ocular conjunctiva (n = 320), skin (n = 92) or peripheral nerve (n = 170) performed between 1975 and 1996, in 486 children. In 112 cases there were definite ultrastructural changes. In 59 cases, the sole ultrastructural exam allowed the diagnosis. In 29, the changes were less specific, and the final diagnosis was performed by a combination of clinical and pathological analysis. In the remaining 24 cases, a generic diagnosis of mucopolysaccharidosis was done in 8 cases, oligosaccharidosis in 4 cases and GM2 gangliosidosis in 12 cases. Whenever a biochemical test was performed in overseas laboratories, the initial diagnosis was confirmed. These results stress the importance of ultrastructural analysis in non-cerebral tissues for the diagnosis of many metabolic disorders mainly when biochemical tests cannot be performed. PMID- 9754424 TI - [Supratentorial meningiomas. Diagnosis, surgical results and complications]. AB - Meningiomas are benign tumors arisising from arachnoid cells and represent the commonest asymptomatic intracranial tumors. We analysed 69 supratentorial meningiomas managed by the Neurosurgical Tumor Group of the Clinics Hospital of Medicine School of Sao Paulo University (September 1995 to September 1997). Age, sex, edema degree, tumor site, surgical complications and mortality were studied. Edema degree was defined by radiological methods (CT and MRI). Forty-seven patients were women and average age was 58 years. Type II of edema degree was predominant (38.7%). Twenty-nine patients had parasagital meningiomas and 40 presented convexity tumors. Simpson I resection was obtained in 48 procedures, II in 18 and III in two surgical removals. Nine cases complicated (transitory deficits, 6; permanent deficit, 1; and infection, 2). Death occurred in two patients. Morbity and mortality had relation with age, falx tumors and attempt of radical surgical removal. Edema degree did not modify mortality and morbidity rates. PMID- 9754426 TI - [Infantile desmoplastic ganglioglioma: a clinical, histopathological and epidemiological study of five cases]. AB - Infantile desmoplastic gangliogliomas are rare tumors of the central nervous system, composed by a mixture of glial and neuronal cells and a fibrous stroma, which affect mainly young patients and arise from the surface of the cerebral hemispheres. We present five cases of infantile desmoplastic ganglioglioma: three were male and two were female. The ages ranged from three months and seven years (mean 2.62 years). The symptoms reflected the growth and topography of the tumors affecting the parietal (n = 3), parieto-occipital (n = 1), occipital (n = 1) lobes. Immunohistochemistry was performed in two cases with similar results to those reported in the literature. PMID- 9754427 TI - [Pituitary apoplexy followed by endocrine remission. Report of two cases]. AB - Pituitary apoplexy is rare and endocrine remission in patients with apopletic secreting pituitary adenomas is even rarer. This study reports on two patients with pituitary macroadenomas (one with Cushing's disease and the other with acromegaly) in whom endocrine remission occurred after apoplexy. The first patient had Cushing's disease and had an ictus of headache and vomiting after which she started a progressive remission of hypercortisolism. A post-apoplexy MRI disclosed persistence of a sellar and supra-sellar mass. She was submitted to transesphenoidal surgery. An hypertensive hemorrhagic cyst was found with no tumor. The second patient had acromegaly. While performing a LHRH-stimulation test he had an ictus of headache, vomiting, no visual loss and appearance of diabetes insipidus. A CT scan disclosed an intrasellar hematoma. Despite the size of the tumor and since there was no visual impairment, this patient was followed up without surgery. Imaging follow-up showed a progressive shrinkage and disappearance of the mass, which was corroborated by endocrine remission. A high rate of recurrence is reported in such patients in the literature. Both patients are being currently followed-up on a long-term basis. PMID- 9754428 TI - [Spontaneous epidural hematoma. Report of two cases]. AB - Spontaneous epidural hematomas are rarely described in literature. They are associated with infectious diseases of the skull, coagulation disorders, vascular malformations of the dura-mater and metastasis to the skull. The authors report two cases of spontaneous epidural hematoma of different etiologies, and study parameters of hemostasis. PMID- 9754429 TI - Lambert-Eaton myasthenic syndrome. Report of two cases. AB - Two cases of Lambert-Eaton myasthenic syndrome, in female patients whose neoplasm investigation was negative, are reported. Repetitive stimulation of ulnar nerve showed an incremental response (+187% and +198%). Needle EMG was normal in one of them, however, the other patient showed fibrillation potentials, positive sharp waves, potentials of low amplitude and short duration. The authors discuss the clinical, electrophysiological, and pathological features of the disease, as well as some aspects of the treatment and follow-up of these patients. PMID- 9754430 TI - Primary lateral sclerosis. A case report with SPECT study. AB - Primary lateral sclerosis (PLS) is a neurodegenerative disease with progressive corticospinal involvement and characterized by lower limbs spasticity followed by upper limbs involvement,rare cranial nerve involvement, typical sparing of all sensory modalities, sphincteric function and eventually mild cognitive changes. The authors report a case of PLS in a 43-year-old woman with 3 years of clinical follow-up and extensive laboratory investigation, including a SPECT study which disclosed bilateral frontal motor area hypometabolism. Several aspects about this unique disease were reviewed,including differential diagnosis with other more common neurological disorders. PMID- 9754431 TI - [3-Hydroxy-3-methyl-glutaryl-CoA-lyase deficiency as coma etiology in the neonatal period: case report]. AB - We report a patient that presented two episodes of coma in the neonatal period, with severe metabolic acidosis and hypoglycemia, without ketosis. The urinary organic acid analysis showed increased amounts of 3-hydroxy-3-methyl-glutaric, 3 methylglutaconic, 3-methylglutaric and 3-hydroxyisovaleric acid. The deficiency of 3-hydroxy-3-methylglutaryl-CoA lyase was diagnosed by the clinical and metabolic features. This disease shows autosomal recessive inheritance and the treatment is done by a diet with restriction of protein (mainly leucine) and lipids, high in carbohydrate content, and the avoidance of fasting and carnitine supplementation. PMID- 9754432 TI - [Fluctuations in Guillain-Barre syndrome related with intravenous human immunoglobulin (triphasic course): case report]. AB - The authors report the case of a patient with severe Guillain-Barre syndrome (tetraplegic and on mechanical ventilation), that was treated with intravenous immunoglobulin (IVIg), 2 g/Kg. At first, there was clinical improvement, followed by clinical deterioration two weeks later. On the second course of IVIg there was, again, clinical improvement and then deterioration, 65 days after treatment. Finally, on the third course of treatment definitive recovery was achieved and no more relapses happened so far (three years after the treatment). The authors review the literature about fluctuations related to treatment with IVIg. Conclusions are that these patients should be closely observed during the first weeks after IVIg treatment, and that further studies are still necessary to elaborate alternative protocols on the prevention of these cases. PMID- 9754433 TI - [Ossification of the posterior longitudinal ligament in the cervical spine: case report]. AB - Ossification of the posterior longitudinal ligament (OPLL) is an uncommon cause of compressive myelopathy in the Caucasian population. A case of spastic paraparesis in a Caucasian man whose radiological investigation showed OPLL is presented. The radiographs of the cervical spine showed a strip of bony density posterior to the vertebral bodies, extending from C2 to T1. Computerized tomography (CT) and CT myelography showed OPLL at the same level. Magnetic resonance showed an area of increased signal on T2-weighted sequences at C7-T1 level suggestive of myelomalacia. The patient underwent an open-door laminoplasty (C2 to C7) with improvement of the paraparesis. OPLL should be included in the differential diagnosis of cervical myelopathy. It can be easily detected by plain radiographs and CT of the cervical spine. A review of the clinical and radiological features and the treatment of OPLL is presented. PMID- 9754434 TI - [Malignant course of a ganglioglioma: case report]. AB - We present the case of a 8-years-old boy, admitted with a history of headache, nausea and vomiting. Cerebral angiography showed a non-vascular mass on frontal lobe. The patient underwent craniotomy and the lesion was removed. Neuropathological study revealed that the tumor was a ganglioglioma. The patient received pos-operative radiotherapy. On follow-up, 16 years after, a computed tomographic scan showed a recurrence of the tumor, and a second surgery revealed a glioblastoma multiform. Gangliogliomas are rare tumors of the central nervous system containing neoplastic ganglion cells and low grade neoplastic glial cells. The malignant degeneration occurs only in the glial component, so the prognosis of these tumors is related to the grade of that component. PMID- 9754435 TI - [Transient ischemic attacks in a patient with superior vena cava obstruction: case report]. AB - The superior vena cava obstruction is a relatively rare condition. We report the case of a 42 year old man suffering of hypertension for about fifteen years. He reported a cervical and thoracic pain for one year, that was related to a 95% of occlusion on the right coronary artery. An angioplasty has been done but the patient still related the thoracic pain. Afterwards the patient had recurrent episodes of right hemiplegia and hypertensive emergencies that have been treated with anti-hypertensive agents. A venous disease was suspected because of cyanosis in the face especially when episodes of transient ischemic attacks occurred. A venography showed obstruction of the right jugular vein near the junction with the superior vena cava. In conclusion, it was not possible to define with certainty the relationship between the two pathologies presented by the patient, even so, we call attention to the improvement of the neurological symptoms after the control of superior vena cava obstruction with the treatment. PMID- 9754436 TI - Experimental HTLV-I infection and associated myelopathy. AB - HTLV-I infection and associated myelopathy has been reproduced experimentally in vitro and in vivo and these studies have shown the possibility of creating several lines of infective cells and of detecting minor and major clinical expressions of HTLV-I associated myelopathy in rabbits and rats. PMID- 9754437 TI - [Vascular dementia: diagnostic challenge and treatment]. AB - BACKGROUND: Treatment of vascular dementia depends on accurate diagnosis and criteria for evaluation of therapeutic responses that are not well standardized. METHODS: Diagnostic criteria for vascular dementia, tools for the assessment of its clinical course and current treatment options are sequentially reviewed. RESULTS: Strict diagnostic criteria with high specificity should be selected in clinical trials. Tools for sequential assessment are not standardized. Clinical endpoints of real value for patients and caregivers are usually excluded from analysis. Prevention of recurrent cerebrovascular events is the only known treatment for stabilization and eventually recovery of cognitive and behaviour disturbances. Benefit may be obtained by adapting hospital or home environment and introducing daily routines minimizing stress and fatigue. The use of non specific cerebral stimulants and so-called neuroprotective drugs is controversial. Careful neuropsychological evaluation guiding management of specific deficits and the use of psychiatric drugs in selected situations may also be useful. CONCLUSION: Strict criteria for the diagnosis of vascular dementia and for the evaluation of treatment responses should be used in drug trials. Aside the secondary prevention of stroke, no drug therapy influencing cognition or neuronal damage has been proved to be useful in patients with vascular dementia. PMID- 9754438 TI - Reduction in BOLD fMRI response to primary visual stimulation following alcohol ingestion. AB - The physiology of alcohol's effects on brain function is poorly understood. Emission tomographic imaging has revealed both acute and chronic alterations in resting cerebral hemodynamics and metabolism following alcohol ingestion. However, cerebral functional integrity under these conditions has received less attention. Functional magnetic resonance imaging (fMRI) offers a non-invasive method for assessing brain functional activation. In order to assess its utility for studying the effect of alcohol on brain function, we performed fMRI with photic stimulation before and after administration of either 0.7 mg/kg alcohol (N = 12) or placebo (N = 5), resulting in peak breath alcohol levels averaging 0.069 g/dl. We found that the amplitude of visual cortical activation in response to photic stimulation was significantly reduced by approximately 33% following alcohol administration (4.0 +/- 1.7% vs. 2.7 +/- 1.3%, P = 0.02), but not following placebo (4.2 +/- 1.5% vs. 4.1 +/- 1.4%, P = 0.7). The results also suggest that the baseline right hemispheric predominance of activation in response to photic stimulation may be reduced following alcohol, suggesting a greater effect on the right hemisphere, consistent with previous studies and alcohol's known effects on visuospatial processing. In addition, through the course of each activation session, there was a progressive reduction in response following alcohol. These data demonstrate that the cerebral effects of alcohol intoxication can be studied with fMRI, and that the effects on brain function of even moderate alcohol intoxication may be widespread, may be lateralized, and may include the visual system. PMID- 9754439 TI - N-[11C]methylspiperone PET, in contrast to [11C]raclopride, fails to detect D2 receptor occupancy by an atypical neuroleptic. AB - The occupancy of the atypical neuroleptic quetiapine (Seroquel) at the D2 dopamine receptor was investigated using the PET tracers [11C]raclopride and N [11C]methylspiperone in a group of five schizophrenic patients. A steady-state treatment condition was ensured by dosing the patients with 750 mg quetiapine daily during 3 weeks followed by a period of tapering off the dose. For each patient, PET examinations were performed with both tracers at two of the following doses: 750, 450, 300 and/or 150 mg. As control, a group of six healthy untreated volunteers was investigated. The D2 binding potential in the putamen and the caudate nucleus was determined by using an evaluation method based on the method proposed by Patlak and Blasberg. The receptor occupancy was determined by assuming that the group of healthy volunteers is representative of untreated drug naive schizophrenic patients. While a significant linear trend of increasing occupancy with increasing quetiapine dose (reaching 51% +/- 10% occupancy at the 750 mg dose) was detected with [11C]raclopride (P < 0.01), no such trend was apparent for N-[11C]methylspiperone (P > 0.09, maximal occupancy values were 2% +/- 3%, measured for the group of three patients on 450 mg). The study suggests that N-[11C]methylspiperone cannot be used for the assessment of D2 receptor occupancy induced by quetiapine. The result is discussed in terms of endogenous dopamine, tracer kinetics and equilibrium dissociation constants. PMID- 9754440 TI - Variations in [3H]imipramine and 5-HT2A but not [3H]paroxetine binding sites in suicide brains. AB - Both the [3H]imipramine and [3H]paroxetine binding sites and the 5-HT2A receptor were simultaneously determined in frontal cortex, cingulate cortex, hippocampus and amygdala from 17 control subjects and 17 depressed suicide victims. A significant decrease in the maximum binding (Bmax) of [3H]imipramine was observed in the hippocampus of suicide victims as compared to control subjects (160 +/- 25 vs. 328 +/- 52 fmol/mg protein; P = 0.007) without changes in the apparent affinity constant (Kd). Furthermore, a significant decrease in the number of 5 HT2A binding sites, together with a significantly lower Kd, was also observed in the hippocampus of suicides as compared to control subjects (129 +/- 18 vs. 225 +/- 32 fmol/mg protein; P = 0.02 and 0.91 +/- 0.07 vs. 1.38 +/- 0.08 nM, respectively; P = 0.006). [3H]Paroxetine binding did not display modifications between the two groups in either Bmax or Kd from any of the brain regions studied. When all four brain regions were taken together, a down-regulation was noted between presynaptic [3H]imipramine binding and the postsynaptic 5-HT2A receptor (r = -0.40; P = 0.0013) in the control group. This correlation did not appear in the suicide group. No correlation was observed between [3H]paroxetine binding and the 5-HT2A receptor in either control subjects or suicides. Taken together, these results suggest that the 5-HT uptake site measured with [3H]imipramine and the 5-HT2A receptors are reliable markers of serotonergic dysfunction. PMID- 9754441 TI - Proton magnetic resonance spectroscopy in acute, juvenile anorexia nervosa. AB - Anorexia nervosa is usually associated with a shrinkage of the brain that is at least partially reversible with weight gain. The pathogenesis of this brain abnormality is unclear. The purpose of this study was to investigate potential alterations in localized proton magnetic resonance (1H MR) spectra of anorectic patients immediately after an interval of excessive weight loss. Twelve patients and seventeen control subjects were examined. Water suppressed 1H MR spectra were recorded from two voxels placed in the thalamus and in the parieto-occipital white matter. The spectra of ten patients could be evaluated. Comparing patients and control subjects, significantly higher signal intensity ratios of choline containing compounds (Cho) relative to total creatine (Cr) as well as significantly lower ratios of N-acetyl-aspartate (NAA) relative to Cho were found in the white matter region. We hypothesize that these results indicate an abnormal starvation, associated membrane turnover, which predominantly takes place in the white matter. No evidence for neuronal degeneration was found in the thalamus or in the white matter region. PMID- 9754442 TI - Quantitative analysis of T2 signal intensities in Alzheimer's disease. AB - Hypointensities (focal areas of decreased signal intensity) have been reported on T2 weighted magnetic resonance images (MRI) in normal aging and in some neurological disease processes. Increased concentrations of iron have been suggested as one cause of these hypointensities. In Alzheimer's Disease, data suggests that there is both a disruption in iron metabolism as well as the presence of T2 hypointensities. We endeavored to determine if the decreased signal intensities could be quantitatively determined and, if so, in what regions, in an effort to establish a non-invasive biological marker and diagnostic aide. We performed a quantitative analysis of the T2 signal intensities in 13 MRIs from AD patients and 16 age- and sex-matched control subjects. We found that while there were statistically significant differences in the intensities of the putamen and red nucleus, these differences were small. We were unable to detect differences in intensities in a whole slice or the frontal lobe. To our knowledge this is the first quantitative comparison of MRI signal intensities in Alzheimer's Disease. PMID- 9754443 TI - On pineal calcification and its relation to subjective sleep perception: a hypothesis-driven pilot study. AB - We classified the degree of pineal calcification (DOC) into seven groups using cranial Computer Tomography (cCT) and then correlated pineal DOC to chronic subjective sleep-related disturbances as measured by a sleep questionnaire in 36 patients. Analysed by logistic regression models, age and sex were not, but higher pineal DOC was significantly associated with the presence of daytime tiredness (OR = 4.15, 95% CI: 1.63, 10.54) and sleep disturbance (OR = 1.74, 95% CI: 1.10, 2.74). This study provides initial confirmation of the hypothesis that the increasing degree of pineal calcification (DOC) might indicate a decrease of melatonin production, which consecutively might lead to a disturbed circadian rhythmicity in the sleep-wake cycle, with the principal symptom being daytime tiredness. PMID- 9754444 TI - The involvement of Bcr in leukemias with the Philadelphia chromosome. AB - In this review, the role of Bcr sequences within Bcr-Abl and the role of the Bcr protein itself in leukemias with the Philadelphia chromosome are discussed. An overview of the oncogenic effects of Bcr-Abl is presented together with Bcr-Abl's effects on several signal transduction pathways. Bcr sequences within Bcr-Abl are known to have an important function in transducing Bcr-Abl's oncogenic signal, yet the role of the Bcr protein encoded by the normal allele is not known. However, several recent findings suggest that Bcr is a conditional negative regulator of Bcr-Abl. From these findings, my colleagues and I have developed a molecular model of the early stage (benign phase) of chronic myelogenous leukemia in which the Bcr protein antagonizes the oncogenic effects of Bcr-Abl. PMID- 9754445 TI - Oncogenes and signal transduction pathways involved in the regulation of Na+ channel expression. AB - Recent progress in neurobiology has revealed that proteins called 'neurotrophic factors' influence development, maintenance of function, and regeneration of neurons in vertebrate nervous system. These factors include the neurotrophin family, epidermal growth factor (EGF), fibroblast growth factor (FGF), and platelet-derived growth factor (PDGF), which are expressed in the nervous system. Effects of the neurotrophic factors are mediated through signal transduction pathways in which several cellular protooncogenes play intrinsic roles. Furthermore, studies on mechanisms coupling membrane events to gene activation have demonstrated that transsynaptic input via action potential and neurotransmitters, and membrane depolarization play an important role in the regulation of electrical activities in neurons during and after maturation. Voltage-dependent sodium (Na+) channels mediate an increase in permeability of Na+ during the initial, rapid phase of the action potential in neurons, and are considered to be important determinants of neuronal functions. Their synthesis and expression, therefore, are crucial aspects of neural differentiation and functions. In mammals, an array of functionally distinct Na+ channels arise, at least in part, through transcriptional regulation of the multiple genes that encode distinct Na+ channel alpha subunits (Na alpha). In this review, we discuss the potential roles of the protooncogenes in the nervous system, with particular emphasis on dynamic expression of the Na+ channel gene family. PMID- 9754446 TI - Immunologic nonresponsiveness to tumors. AB - Over the past several years it has become clear that malignant cells express a variety of tumor associated antigens, and T cells reactive to these antigens have been identified. However, the T cells are not effective in rejecting tumors. In general, T cells that are not tolerized within the thymus have the potential to be rendered tolerant by one of three mechanisms. Immune deviation occurs when regulatory T cells which share a common precursor differentiate away from the phenotype required to effect a particular immune response. Anergy induction occurs when a T cell is stimulated through its T cell receptor in the absence of costimulation. Activation-induced cell death (AICD) is apoptosis of activated T cells upon subsequent encounter with antigen. There is emerging information that some of these mechanisms can be responsible for the lack of T cell responsiveness to tumor cells. Also, tumor cells can acquire attributes that interfere with an immune response, including down-regulation of MHC molecules or other molecules involved in antigen processing; secretion of the immunosuppressive cytokine TGFbeta; and expression of the apoptosis-inducing surface molecule, Fas ligand. An expansion in our understanding of how tumor cells evade a T cell mediated death will provide insight into potential strategies to improve immunotherapeutic approaches to cancer patients. PMID- 9754447 TI - The c-Fes family of protein-tyrosine kinases. AB - The human c-fes protooncogene encodes a protein-tyrosine kinase (c-Fes) distinct from c-Src, c-Abl and other nonreceptor tyrosine kinases. Although originally identified as the cellular homolog of several transforming retroviral oncoproteins, Fes was later found to exhibit strong expression in myeloid hematopoietic cells and to play a direct role in their differentiation. Recent work has shown that Fes exhibits a more widespread expression pattern in both developing and adult tissues, suggesting a general physiological function for this kinase and its closely related homolog, Fer. This review highlights the unique aspects of Fes structure, regulation, and function that set it apart from other tyrosine kinase families. PMID- 9754448 TI - Enteropathy-associated T-cell lymphoma and its immunocarcinogenic correlates: case report and review of the literature. AB - We report a case of enteropathy-associated T-cell lymphoma (EATL), diagnosed by small intestine and gastric biopsies, who presented with manifestations of hypocalcemia and malabsorption. Immunological assessment revealed increased expression levels of tumor necrosis factor system components and eotaxin, an observation that is consistent with the cytotoxic T-cell phenotype characteristic of EATL, and decreased numbers of circulating activated (CD8+CD38+ and CD4+CD25+) and suppressor (CD11b+) T cells, a feature which can contribute to lymphomagenesis in patients with celiac disease. The acute clinical presentation of the patient resolved with mineral and vitamin supplementation and a gluten free diet. The novel immunological findings described are discussed in the context of a review of our current knowledge of the immunopathogenesis of celiac disease and associated intestinal neoplasia. PMID- 9754449 TI - Mechanisms of p53-mediated apoptosis. AB - p53-mediated apoptosis of cells with DNA damage or oncogene overexpression is a major mechanism for its function as a tumor suppressor. Both transcriptionally dependent and transcriptionally independent activities of p53 can play a role in mediating cell death. It appears that p53 can induce apoptosis by multiple pathways in a manner which is regulated in a cell type and signal-specific fashion. Understanding the biochemical mechanisms of p53-dependent apoptosis holds a promise of manipulating these pathways in cancer therapy. PMID- 9754450 TI - Functional magnetic resonance imaging of alprazolam-induced changes in humans with familial alcoholism. AB - This study sought to identify whether subjects with a family history (FH + ) of alcoholism had changes in regional cerebral blood volume (rCBV) after an alprazolam challenge which distinguished them from subjects without a family history (FH -) of alcoholism using functional MRI (fMRI). Twelve FH + and eight FH - subjects were challenged with 1 mg of alprazolam or placebo in a double blind crossover design. FMRI scans were obtained at baseline, 1 and 2 h after the challenge using the dynamic susceptibility contrast method with gadolinium. Mood scales, the Tufts Addiction Research Center Inventory-Morphine Benzedrine Group Scale and the drug liking scale, were administered every 30 min to assess drug effects. Global analysis of CBV showed a treatment by time decrease on alprazolam relative to placebo, but no effect by family history. The FH + group showed rCBV decreases at 1 h in the left caudate and left inferior prefrontal region, while the FH - group showed rCBV decreases at 2 h in the right inferior prefrontal region and anterior cingulate in response to alprazolam relative to placebo. FH + subjects reported more mood enhancement with alprazolam. This fMRI technique detected global and regional CBV changes induced by alprazolam. The location and rate of alprazolam-induced rCBV changes differed between FH + and FH - subjects. These changes may be related to the increased mood enhancement found in subjects genetically predisposed to alcoholism. PMID- 9754451 TI - Functional connectivity in depressive, obsessive-compulsive, and schizophrenic disorders: an explorative correlational analysis of regional cerebral metabolism. AB - In order to investigate the changes in functional relationships between brain regions in three psychiatric disorders, an exploratory statistical analysis of the regional cerebral metabolic rates for glucose (rCMRglu) obtained with positron emission tomography (PET) was performed. Correlations between various rCMRglu were computed in control, depressive, obsessive-compulsive, and schizophrenic groups to determine whether alterations of the correlation pattern were found in the psychiatric groups. The absence of correlation between left and right frontal lobes was common to the three psychiatric groups studied. Depressive patients recovered a better frontal interhemispheric coupling after successful treatment and the alterations in the depressed state also involved the relation between the right temporal cortex and the right thalamus. Obsessive compulsive patients had not only lateral frontal dysfunction but also alterations in the functional relationships between cortex and thalami. In schizophrenic patients, the modifications of regional cerebral metabolic correlations involved both anterior and posterior cortical regions. Thus, although the relationship between left and right frontal lobes was altered in three psychiatric disorders, the pattern of functional connectivity between frontal regions, posterior cortical and subcortical regions differed depending on the diagnostic group. PMID- 9754452 TI - Metabolic brain mapping in Alzheimer's disease using proton magnetic resonance spectroscopy. AB - Alzheimer's disease (AD) is a progressive disorder associated with disruption of neuronal function and neuronal loss. N-acetylaspartate (NAA) is a marker of neuronal content and can be assessed using proton (1H) magnetic resonance spectroscopy (MRS). We utilized 1H-MRS (two-dimensional chemical-shift imaging) to assess amplitudes and areas of NAA, as well as choline moieties (Cho), creatine (Cr) and myo-inositol (mI), in 15 AD patients compared with 14 control subjects. Voxels were classified as predominantly cortical gray matter (CGM), subcortical gray matter (SGM), or white matter (WM). Compared with control subjects, AD patients exhibited decreased NAA/Cho and NAA/Cr amplitudes, whereas an increase was observed in Cho/Cr and in amplitude ratios involving mI. Area ratios were significant in the same direction for NAA/Cho, NAA/Cr, mI/Cr and mI/NAA. No significant effects of tissue type were observed; however, significant group x tissue type interactions were noted for Cho/Cr and mI/Cr amplitudes. Our study confirms that 1H-MRS can identify distinct physicochemical alterations in AD patients, reflecting membrane changes and diminished neuronal function. These alterations can be used as longitudinal markers for the disease. PMID- 9754453 TI - Central effects of acamprosate: part 1. Acamprosate blocks the glutamate increase in the nucleus accumbens microdialysate in ethanol withdrawn rats. AB - One of the known behavioral actions of acamprosate is to decrease hypermotility during alcohol withdrawal. However, the mechanism of this effect remains unclear. In this study, the concentrations of excitatory and inhibitory amino acids were assayed by the microdialysis technique with OPA/BME precolumn derivatization and electrochemical detection in the nucleus accumbens of male Wistar rats which were either alcoholized by ethanol inhalation or simultaneously alcoholized and treated orally by acamprosate (400 mg/kg/day) for 4 weeks. Without treatment, extracellular glutamate increased during the withdrawal phase, while other amino acids tested (aspartate, arginine, taurine, alanine and GABA) remained stable. In contrast, the alcoholized rats treated with acamprosate failed to present the increase in glutamate during ethanol withdrawal, while other amino acids tested also remained stable. The observed glutamate increase could be responsible for the hyperexcitability observed during episodes of ethanol withdrawal. These results suggest that acamprosate is able to reduce the ethanol withdrawal syndrome by reducing the concentration of glutamate in the nucleus accumbens. PMID- 9754454 TI - Central effects of acamprosate: part 2. Acamprosate modifies the brain in-vivo proton magnetic resonance spectrum in healthy young male volunteers. AB - Although acamprosate is a drug which is successfully used for therapy in maintaining alcohol abstinence following alcohol withdrawal in chronic alcoholism, little is understood about its mechanism of action in the central nervous system. Our objective was to assess the effects of acamprosate on the central nervous system in healthy subjects by dynamic proton magnetic resonance spectroscopy (MRS) measurements localized in brain tissue in vivo. Recordings were performed after intravenous administration of acamprosate or placebo to eight healthy male volunteers participating in a double-blind, randomized, cross over, placebo-controlled study. The data were acquired using a spin-echo volume selective localized spectroscopy scheme on a 3-T whole body MRS system. Spectra obtained at baseline and at 20-min time intervals after the beginning of drug infusion were analyzed on the basis of five non-overlapping spectral integration regions. In the acamprosate-treated group, the median integral values in the regions for which N-acetylaspartate and glutamate are the main signal contributors showed decreases relative to placebo 20 min after the infusion began. Results suggest a central glutamatergic effect of acamprosate consistent with cerebral microdialysis glutamate measurements in vivo obtained from alcoholized rats treated with acamprosate (Part 1 of this study). This study is to our knowledge the first one describing a central effect of acamprosate in humans by MRS. PMID- 9754455 TI - Asymmetrical changes in blood-brain barrier permeability during pentylenetetrazol induced seizures and in acute hypertension. AB - The asymmetrical breakdown of the blood-brain barrier to Evans-blue was studied in male and female rats during epileptiform seizures and in acute hypertension. The animals were divided into six groups. Group I: control female; Group II: control male; Group III: female + acute hypertension; Group IV: male + acute hypertension; Group V: female + seizure; Group VI: male + seizure. Asymmetric breakdown of the blood-brain barrier had been seen in female rats treated with pentylenetetrazol. Pentylenetetrazol-induced seizure produces less disruption of the blood-brain barrier in right cerebral hemisphere than in left cerebral hemisphere in female rats. There were no asymmetrical changes of blood-brain barrier permeability between the left and right hemispheres in acute hypertension in both sexes, and male rats treated with pentylenetetrazol. PMID- 9754456 TI - Clinical use of profiled hemodialysis. AB - The new population on dialysis today consists mainly of high risk patients (the elderly, diabetics, etc.) with high cardiovascular scores, and such vascular pathology is the most important predisposing factor for the occurrence of a frequent intradialytic clinical complication, vascular instability syndrome, which covers a range of clinical problems. Recently a new dialysis technique, profiled hemodialysis (PHD), has been set up and proposed for routine use. PHD consists of the clinical use of preestablished individual dialysis profiles aimed at antagonizing the changes in intradialytic plasma osmolarity by continuous modulation of dialysate sodium concentration throughout the whole extracorporeal session. In particular, PHD aims at reducing the fall of plasma osmolarity in the first half of the session (when it is higher) by reducing the sodium removal rate through increasing its dialysate concentration while taking into account the desired individual sodium balance to be reached at the end of the session. In this work, we report clinical experience with PHD compared to standard hemodialysis with constant sodium dialysate (SHD) in terms of its efficacy to maintain a more stable intradialytic blood volume (BV) and more stable hemodynamics. The PHD used in this work has been implemented by a mathematical model for computing the individual dialysate sodium profile which we have recently validated (Ursino M, Coli L, La Manna G, Grilli Cicilioni M, Dalmastri V, Guidicissi A, Masotti P, Avanzolini G, Stefani S, Bonomini V. A simple mathematical model of intradialytic sodium kinetics: "in vivo" validation during hemodialysis with constant or variable sodium. Int J Artif Organs 1996;19:393 403.). Eleven uremic patients affected by hypotension at the beginning of dialysis treatment were studied. Each patient first underwent an SHD treatment and 1 week later a PHD treatment. The 2 extracorporeal sessions (one on SHD and the other on PHD) were performed in each individual patient under identical operative conditions including the sodium mass removal by the end of the session and the ultrafiltration rate. The crit line and Doppler echocardiography were used to determine BV, cardiac output (CO), and stroke volume (SV) throughout the sessions. The mean blood pressure (MBP) and heart rate (HR) were simultaneously monitored. PHD was associated with a more stable intradialytic BV and more stable hemodynamics compared to SHD. The higher stability of BV and cardiac function (in terms of SV and CO maintenance) which was obtained above all in the first half of the PHD session was associated with a higher stability of the MBP and the HR. This resulted in an enhancement in cardiovascular tolerance to ultrafiltration throughout the session in all tested patients. In contrast, SHD in the same patients was characterized by early significant changes in BV and cardiovascular parameters resulting in a significant decrease of the MBP and a significant increase of the HR throughout the session and also 1 h after the end of dialysis. Our results indicate that PHD may represent an efficient approach for the treatment of patients suffering from intradialytic vascular instability. If long term clinical practice confirms the efficacy of PHD in controlling dialysis intolerance symptoms, it will have great scope as a routine procedure. PMID- 9754457 TI - Prediction of anticoagulation during hemodialysis by population kinetics and an artificial neural network. AB - All systems currently used for routine hemodialysis require heparin administration to prevent blood clotting in the extracorporeal circuit. We tested the hypothesis that population-based statistical techniques can be used to predict heparin concentrations during routine hemodialysis. Two predictive models were developed, one based on nonlinear mixed effects modeling (NONMEM) and the other on a multilayer perceptron neural network. Serial clotting time data were obtained from forty-nine patients and used to develop the models. The models were used to predict the clotting times of 70 patients in a prospective test. We determined that the neural network provided greater precision, had fewer outliers in its predictions, and did not have the model misspecification in bolus administration that the NONMEM predictions demonstrated. A final NONMEM model was developed using all data from 119 patients to identify important covariates for predicting the heparin pharmacodynamic parameters, volume of distribution, and clearance. Both the volume of distribution and clearance increased following the initiation of dialysis and as the patient's baseline clotting time increased. The volume of distribution also increased as the patient's weight increased but was decreased by smoking and diabetes. Population-based statistical techniques may provide a useful alternative to existing methods for prescribing heparin. PMID- 9754458 TI - Left ventricular mass regression after implantation of St. Jude Medical cardiac valves in small aortic roots. AB - In this study, we analyzed the extent of regression of left ventricular hypertrophy in patients who received small St. Jude Medical (SJM) aortic valves and compared the results with those of another group receiving larger valves. Eighty-eight patients received either 19 or 21 mm valves (Group 1, 25 patients) or either 23 or 25 mm valves (Group 2, 53 patients). Echocardiographic studies were done before the operation and 5 years postoperatively. At follow-up a significant reduction in the left ventricular mass was found for both patient groups (p < 0.0001). Doppler echocardiography derived pressure gradients for both groups were obtained during the follow-up period. As expected, the patients in Group 1 had higher peak pressure gradients than did those in Group 2. However, there was no significant difference between the 2 groups or any significant correlations between peak pressure gradients and body surface area (BSA). Actuarial survival was 84.7% at 15 years for Group 1 and 85.9% at 17 years for Group 2. Actuarial freedom from valve related events was 91.4% at 15 years for Group 1 and 82.7% at 17 years for Group 2. There was no significant difference in survival or valve related event free curves between the 2 groups. After implantations of SJM valves in small aortic roots, significant left ventricular mass regression was obtained, and the results were comparable to those for valves of other sizes. The long-term performance of aortic valve replacement with small valves was satisfactory as judged by improvement in the functional class of patients and survival statistics, the durability of the prosthesis, and valve related morbidity comparable to that of valves of other sizes. PMID- 9754459 TI - A continuous and pulsatile flow circulation system for evaluation of cardiovascular devices. AB - The design of a nonpulsatile and pulsatile system using a centrifugal pump is presented. To induce a pulsatile flow with a centrifugal pump, an independent pneumatically driven unit provided flow patterns over a wide range of frequencies and amplitudes. The pulsatile flow was generated by the axial displacement of a cylinder that periodically compressed the flexible conduit that is connected to the pump. The system can accommodate flow rates up to 6,000 ml/min and transmural pressures up to 500 mm Hg and is capable of maintaining the pressure at a constant value. This circuit produced reproducible pressure waves having a frequency up to 4 Hz. The periodicity of the transmural pressure between 80 and 180 mm Hg was similar to the pressure wave propagation observed in peripheral circulation. Capable of adequately reproducing continuous and pulsatile flow, the apparatus is therefore versatile to allow in vitro evaluation of cardiovascular devices. PMID- 9754460 TI - Evaluation of platelet adhesion and activation on materials for an implantable centrifugal blood pump. AB - A totally implantable centrifugal artificial heart has been developed in which a pivot bearing supported centrifugal pump is used as a blood pump. The following have been adopted as blood contacting materials in our pump: titanium alloy (Ti 6A1-4V) for the housing and impeller, alumina ceramic (Al2O3) for the male pivots, and ultrahigh molecular weight polyethylene (PE) for the female pivots. Greater antithrombogenicity is required for an implantable blood pump. To examine the thrombogenicity of these materials, we evaluated in vitro platelet adhesion and activation, which may play key roles in thrombogenesis on foreign surfaces. Ti-6A1-4V, Al2O3, and PE were compared with polycarbonate (PC), silicone carbide (SiC), and pure titanium (pTi). Platelet adhesion was assessed using monoclonal antibody (CD61) directed against glycoprotein IIIa. Platelet activation was evaluated by measuring P-selectin (GMP-140) released from irreversibly activated platelets. Each material with a surface area of 16.6 cm2 was incubated with 2.5 ml of plasma or 2.5 ml of heparinized fresh whole blood for 3 h at 37 degrees C. The optical density (OD) at a wavelength of 450 nm for CD61 was 0.93+/-0.35 in PC, 0.34+/-0.13 in PE, 0.27+/-0.13 in pTi, 0.26+/-0.01 in Al2O3, 0.21+/-0.04 in SiC, and 0.12+/-0.12 in Ti-6A1-4V. The GMP-140 levels of the tested materials were not significantly different from the control value (45.9+/-7.2 ng/ml). These results indicate that Al2O3, PE, and Ti-6A1-4V, which are incorporated into our implantable centrifugal pump, have satisfactory antithrombogenic properties in terms of platelet adhesion. However, platelet activation by any material was not observed under the static condition in this study. PMID- 9754461 TI - Minimally invasive cardiac surgery: current status and perspective. AB - Recently, the minimally invasive approach has become a growing aspect in the field of cardiac surgery with the goal of eliminating cardiopulmonary bypass (CPB) and/or median sternotomy. In coronary bypass surgery, the application of this approach is direct anastomosis, primarily of the left internal thoracic artery to the left descending coronary artery under a beating condition without the use of CPB through a small left thoracotomy minimally invasive direct coronary artery bypass (MIDCAB). In the repair of intracardiac lesions, CPB cannot be excluded, but a small right parasternal incision or small partial sternotomy (ministernotomy) has been applied for congenital defects and mitral and aortic valve lesions. With technological advances in CPB, these approaches may become more popular in the near future. PMID- 9754462 TI - Minimally invasive cardiac surgery in the adult: surgical instruments, equipment, and techniques. AB - To clarify the special instruments and equipment used for minimally invasive cardiac surgery (MICS), we examined the initial experiences with MICS operations with ministernotomy or minithoracotomy at our institution. Fifty adult patients with congenital, valvular, and/or ischemic heart diseases underwent MICS operations, and all surgical procedures were completed without conversion to full sternotomy. The length of the skin incision was about 10 cm or less in all patients. Postoperative recovery was favorable, and the majority of the patients were discharged from the hospital around the end of the second postoperative week. In this series of patients, an oscillating bone saw, lifting type retractor, 2 blade spreader, cannula with a balloon, and right-angled aortic clamp among other items, were very useful for successfully performing various operations with MICS approaches and techniques. The associated results suggest that MICS with ministernotomy or minithoracotomy was feasible using special instruments and equipment and could be encouraged for adult patients with various cardiovascular diseases. PMID- 9754463 TI - Less invasive coronary artery bypass without cardiopulmonary bypass. AB - Minimally invasive coronary artery bypass grafting (CABG) aims to avoid cardiopulmonary bypass and take maximum advantage of a smaller incision. Minimally invasive direct coronary artery bypass (MIDCAB) surgery is performed on selected arteries of the beating heart under direct vision through a choice of small incisions. Short-term results show good patency rates and a dramatic impact in terms of shorter hospital stays and cost effectiveness. The procedure is also being used increasingly in Japan. However, valid concerns have been raised about the quality of the anastomosis fashioned on a beating heart with pharmacologic bradycardia, and the long-term result of this technique is still questionable. The combined use of circulatory assist devices and mechanical stabilizing devices will be expected to expand access to coronary arteries by allowing for decompression of the left ventricle, permitting retraction and rotation of the heart, and hopefully further improvement of the results. Less invasive coronary surgery should be proven to be as effective and safe as conventional CABG before widespread adoption. PMID- 9754464 TI - Alteration of the traditional extracorporeal bypass circuit to accommodate port access minimally invasive cardiac procedures using endovascular based cardiopulmonary bypass. AB - Port-Access minimally invasive cardiac surgery systems (Heartport, Inc., Redwood City, CA, U.S.A.) enable surgeons to perform many procedures including valve surgery and complete coronary artery revascularization of all surfaces of the heart through small anterior thoracotomies. The endovascular based EndoCPB (Heartport, Inc.) cardiopulmonary bypass system uses a modified extracorporeal circuit to afford the same level of myocardial protection through cardioplegic cardiac arrest and bypass as is provided in traditional open chest surgery. We describe the changes required to convert a conventional CPB pump circuit to perform Port-Access procedures and make recommendations based on clinical experience to facilitate establishing a Port-Access surgical team and interpreting EndoCPB pressure and flow data. Specific emphasis is placed on the expanded role of the perfusionist in these cases. PMID- 9754465 TI - Acetate hemodialysis does not increase the frequency of arrhythmia in hemodialysis patients. AB - Arrhythmia is one of the most important causes of mortality in patients on hemodialysis and may develop due to cardiovascular diseases or fluid-electrolyte or acid-base abnormalities. Previous studies have shown that acetate hemodialysis (AHD) increased the frequency of arrhythmia. To evaluate the frequency and the causes of arrhythmias during AHD, we studied 33 randomly selected patients (25 male and 8 female, mean age of 45+/-18 years) who were under AHD (4 h, 3 times/week, mean duration of HD of 38+/-29 months) with the same Cuprophan membranes. All patients underwent a detailed echocardiographic evaluation during the interdialytic period. Twenty-four hours of Holter monitoring was performed starting from the onset of HD. Twelve lead electrocardiography (ECG) was obtained, and venous and arterial blood samples were drawn for serum electrolytes, pH, and arterial blood gas measurements before and after HD. Serum magnesium and potassium levels dropped after AHD (from 2.3+/-0.5 to 1.9+/-0.3 mEq/L and from 5+/-0.7 to 3.4 +/-0.4 mEq/L respectively, p < 0.001); on the other hand serum pH, bicarbonate, sodium, and calcium levels were normalized. Electrocardiographic evaluation revealed significant lengthening of the QTc interval (from 433+/-42 to 464+/-43 ms, p < 0.001), which was thought to be related to the decrease in serum magnesium and potassium levels. The frequencies of ventricular premature contractions (VPCs) were not different during AHD and the interdialytic period (8+/-9.1 to 6.5+/-11 contractions/h, p > 0.05). This was also true for supraventricular premature contractions (SVPCs) and supraventricular tachycardia (SVT). Nonsustained ventricular tachycardia was observed in 2 patients during HD and in 1 patient in the interdialytic period. No relation was established between the echocardiographic findings and the frequency of arrhythmia. In our ambulatory electrocardiographic study, the frequencies of VPCs and SVPCs observed during the interdialytic period were only positively correlated with age (r=0.54, p=0.013 and r=0.50, p=0.010, respectively). No relation was found between the frequency of arrhythmia and the gender of the patients; duration of HD; etiology of kidney disease; or serum Na, K, Ca, iCa, Mg, bicarbonate, or pH levels (p > 0.05). In conclusion, the application of AHD does not increase the frequency of arrhythmia in HD patients as had been shown in previous studies. PMID- 9754466 TI - Peritoneal clearance and peritoneal transfer of oxalic acid, vitamin C, and vitamin B6 during continuous ambulatory peritoneal dialysis. AB - The peritoneal clearance and peritoneal transfer of oxalic acid, vitamin C, and vitamin B6 in 32 patients during continuous ambulatory peritoneal dialysis (CAPD) using peritoneal dialysis solutions containing 1.5% or 2.5% glucose were examined. The plasma level of oxalic acid was significantly elevated in all patients, plasma vitamin C was in the normal range or in the upper margin of the normal range, and plasma vitamin B6 was in the normal range. The peritoneal clearance of oxalic acid was significantly lower, and the peritoneal clearance of vitamin B6 was the lowest in comparison to the peritoneal clearance of urea. With the exception of vitamin B6, the peritoneal clearance and peritoneal transfer of the examined parameters increased using the dialysis solution containing 2.5% glucose. We found direct relationships between the plasma levels of oxalic acid and creatinine as well as plasma vitamin C and between the peritoneal transfer of oxalic acid and the peritoneal transfer of vitamin C as well as vitamin B6. The significant hyperoxalemia of our patients was found to persist despite the relatively high peritoneal transfer of oxalic acid during CAPD. These results suggest that CAPD is not a method effective enough for permanent reduction of the plasma levels of oxalic acid. PMID- 9754468 TI - Retrieval analysis of mechanical heart valves: impact on design and clinical practice. AB - Explanted mechanical heart valves were examined nondestructively, and the findings were related to guidelines, technical reports, and other information to judge the risk of failure and its possible impact on valve design and clinical practice. Diagnoses for single valves could be made, but risks and rates of failure for patient populations could not be predicted due to insufficient information concerning the manufacturing process and valve and patient numbers. Based on the results of this study and the principle that decisions on recalls and patient counseling must be based on scientific knowledge rather than on wait and see policies, the following is recommended: registration of all implanted valves, follow-up of a large cohort of valve carriers, comparison of wear test results of preimplant and postretrieval valves, maintenance of a reference stock of valves and materials, and submission of failure scenarios to certifying bodies. PMID- 9754467 TI - Membrane immunoisolation of a diffusion chamber for bioartificial pancreas. AB - Immunoisolation is a potentially important approach to transplanting islets without any immunosuppressive therapy. The concept of immunoisolation is outlined in systems in which the transplanted tissue is separated from the immune system of the host by an artificial barrier. We previously described a diffusion chamber as a bioartificial endocrine pancreas (Bio-AEP), which was constructed by placing pancreatic endocrine cells, trapped in a mixed matrix, in the center of a ring holder sandwiched between nucleopore membranes, which were shielded by silicone. This experiment was designed to evaluate a suitable pore size for the nucleopore membrane to ensure immunoisolation during xenoimplantation of the Bio-AEP in vitro and in vivo. A nucleopore membrane of pore size 0.1 microm or 0.2 microm was employed as the semipermeable membrane which provided a mechanical barrier between the endocrine pancreas graft and the host immune system. The protective effect of the Bio-AEP from humoral immunity was determined in vitro, using sensitized sheep erythrocytes (EAs). A complement protein did not destroy the cell membranes of the EAs in the diffusion chamber containing the mixed matrix with the nucleopore membrane of 0.1 microm pore size. In an in vivo experiment, 6 streptozotocin (STZ) induced diabetic rats were implanted with Bio-AEPs constructed with nucleopore membranes of pore size 0.1 microm and containing MIN6 cells in the mixed matrix. In the STZ diabetic rats with Bio-AEPs, a return to normoglycemia was observed up to 50 weeks after implantation without the use of any immunosuppressant. Also, the body weights of the rats gradually increased. During the observation, when the Bio-AEPs were removed from the STZ diabetic rats, the blood glucose immediately returned to preimplantation levels, and the body weights of the rats also decreased. The membranes of the Bio-AEPs removed from the STZ diabetic rats showed a very thin layer of fibroblastic cells on the outer surfaces. The results indicated that the Bio-AEP, in which pancreatic endocrine cells were trapped in a mixed matrix and with a 0.1 microm pore size membrane, should be useful for xenoimplantation into diabetic animals and may open a new field in the therapy of human diabetics. PMID- 9754469 TI - Assessment of bovine platelet life span with biotinylation and flow cytometry. AB - Reduced platelet life span is associated with the implantation of a variety of cardiovascular devices and may be used as a gauge of device biocompatibility. In the bovine model, platelet life span has previously been assessed with radioisotope labeling of removed platelets followed by reinjection and periodic gamma counting of blood samples. We report here the use of protein-reactive biotin (sulfo-N-hydroxysuccinimido [NHS]-biotin) as an alternative to radioisotope techniques whereby reinjected biotinylated platelets are subsequently detected in blood samples using phycoerythrin-streptavidin and flow cytometric techniques. Platelet life span was quantified in a normal calf (4.9 days) and in a calf prior to (6.1 days) and following (3.1 days) implantation of a Nimbus Axial Flow Pump ventricular assist device. The assessment of bovine platelet life span with biotinylation and flow cytometry avoids the technical, regulatory, and safety considerations associated with radioisotope usage and appears readily amenable to application in cardiovascular device testing. PMID- 9754470 TI - Effect of a balanced biventricular bypass system on left ventricular energies. AB - Left dominant biventricular failure is a common type of heart failure after cardiac surgery. We developed a biventricular bypass (BVB) system for treatment of postcardiotomy ventricular failure, and we previously reported that the clinical results of the BVB system were superior to those obtained with venoarterial bypass (VAB). The purpose of this study was to evaluate the effect of the BVB system on left ventricular (LV) performance in comparison to that of VAB by means of the LV pressure-volume relationship (PVR). Eight adult mongrel dogs (14-21 kg) underwent VAB with right atrial and aortic cannulation. Left atrial cannulation was added for BVB, and both atrial drainage tubes were joined with a Y-shaped connector. The bypass flow was maintained at half of the baseline cardiac output (0.7-1.0 L/min), and the hemodynamic parameters were monitored. A high fidelity microtip catheter and a conductance catheter were used to evaluate LV function. The slope of the LV end-systolic pressure-volume relation (Emax), the slope of the LV end-systolic pressure-stroke-volume relation (Ea), the LV stroke work (SW), LV potential energy (PE), LV pressure-volume area (PVA), the slope of the SW end-diastolic volume relation (PRSW), and an index of the LV energizing charge (ratio of PE/PVA) were assessed during transient occlusion of the inferior vena cava. LV contractility (Emax) showed no significant change during each experiment. Standardized LV work (PRSW) was reduced by BVB in comparison to the baseline and in comparison to VAB. The rate of LV energy charge (PE/PVA) significantly increased only during BVB. These results suggested that the BVB system might be an effective circulatory support for reducing LV work and improving the LV energizing charge in patients with severe heart failure after cardiac operation. PMID- 9754471 TI - Type II SLAP lesions: three subtypes and their relationships to superior instability and rotator cuff tears. AB - One hundred two type II SLAP lesions without associated anterior instability, Bankart lesion, or anterior inferior labral pathology were surgically treated under arthroscopic control. There were three distinct type II SLAP lesions based on anatomic location: anterior (37%), posterior (31%), and combined anterior and posterior (31%). Preoperatively, the Speed and O'Brien tests were useful in predicting anterior lesions, whereas the Jobe relocation test was useful in predicting posterior lesions. Rotator cuff tears were present in 31% of patients and were found to be lesion-location specific. In posterior and combined anterior posterior lesions, a drive-through sign was always present (despite absence of anterior-inferior labral pathology or a Bankart lesion) and was eliminated by repair of the posterior component of the SLAP lesion. We conclude that SLAP lesions with a posterior component develop posterior-superior instability that manifests itself by a secondary anterior-inferior pseudolaxity (drive-through sign), and that chronic superior instability leads to secondary lesion-location specific rotator cuff tears that begin as partial thickness tears from inside the joint. PMID- 9754472 TI - Hi-frequency electrical cautery stimulation in the treatment of displaced meniscal tears. AB - This is a preliminary report of four cases of meniscal displaced tears: two bucket handle medial meniscus tears, one complex tear in the avascular zone or lateral meniscus, and one longitudinal full-thickness tear on the lateral meniscus. These tears were treated by applying hi-frequency current stimulation to the tissues and obtaining total meniscal visual healing on a second arthroscopic view after 6 weeks. PMID- 9754473 TI - Ankle joint arthroscopy for meniscoid lesions in athletes. AB - The meniscoid lesion is a frequent but not well known cause of persistent pain in the anterior part of the upper ankle in sports traumatology. It has been described as portions of hyalinized tissue following an inversion sprain of the ankle. Trapping of this formation between the lateral cheek of the talus and the fibula is supposed to be responsible for pain and other symptoms reported by the patient. In 59 arthroscopic procedures on the ankle joint in athletes, meniscoid lesions were seen in 19 cases. Only 1 of these 19 patients showed lateral and anterior instability, and frequent clinical symptoms were swelling and trapping. Intraoperatively, all meniscoid lesions were combined with synovitis. Chondromalacia and osteophytes were seen several times. After an average follow up period of 12 months, 14 patients could be examined. Twelve of the athletes returned to full sports activity; 10 were very satisfied, 2 satisfied, and 2 unsatisfied. Nine patients did not complain of any swelling, 4 did so on rare occasions, and 1 complained persistently. No pain was reported 10 times, improvement of pain 3 times, and continuing persistent pain 1 time, probably because of simultaneous chondromalacia and osteophytes. These were found more frequently in patients with a longer case history and unsuccessful conservative treatment, so that early arthroscopic surgery is recommended. PMID- 9754474 TI - Spontaneous vacuum pneumarthrography revisited: the significance of the vacuum phenomenon in the lateral compartment of the knee. AB - We report seven cases of spontaneous vacuum phenomenon of the lateral compartment of the knee. Previous reports of the vacuum phenomenon suggest that it is related to traction on a joint or the absence of an effusion. The presence of this finding on plain radiographs, or artifacts associated with it on magnetic resonance imaging, have been said to create a false-positive indication for meniscal tears, especially in the medial compartment. We describe plain radiographic, magnetic resonance, and arthroscopic findings in seven consecutive patients with vacuum phenomenon of the lateral compartment of the knee and conclude that this finding may be a true-positive indication for degenerative meniscal tears and chondrosis. The etiology of this finding and its long-term significance remains unclear. PMID- 9754475 TI - Changes in muscle strength properties caused by harvesting of autogenous semitendinosus tendon for reconstruction of contralateral anterior cruciate ligament. AB - A prospective study was conducted of how the muscle strength of the donor knee is affected by harvesting of the autogenous semitendinosus tendon (St) for use as a substitute graft material in cruciate ligament reconstruction. There were 25 patients from whom only the St was harvested from the contralateral (i.e., healthy/donor) knee. Using a Biodex System II (Biodex, New York, NY), the strength of the donor knee was measured during both extension and flexion, both before and 12 months after the tendon harvesting procedure. A comparative study was made of the preharvest and postharvest values for the peak torque and peak torque angle in the isokinetic contraction. There were no statistically significant differences between the preharvest and postharvest peak torque values of the donor knee. However, the peak torque angle decreased significantly after the tendon harvest; the range of the mean decrease was from 11.7 degrees to 15.0 degrees. This indicates that there was a change to a small flexion angle (P < .05). After the tendon harvest, regardless of the applied angular velocity, more than 80% of the cases showed a change of torque curve shape in which there was no peak in the latter half, and the position of the peak was shifted to the left. In conclusion, the results of this study indicate that harvesting of the autogenous St does not affect the peak torque, but the peak torque angle during flexion of the donor knee is reduced. PMID- 9754476 TI - Radiofrequency (electrosurgical) ablation of articular cartilage: a study in sheep. AB - The objective of this study was to examine the effect of a bipolar ablation probe on experimentally roughened articular cartilage and compare it with the traditional mechanical shaving technique using the knee joint of sheep. Twenty eight skeletally mature ewes were divided randomly into two groups: one group was treated with a rotating shaving device and another group was treated using the bipolar ablation probe (Bipolar Arthroscopic Probe; Electroscope, Inc, Boulder, CO). Animals were killed at 0, 6, 12, and 24 weeks, and histological sections of the experimental limbs were compared with sections of the opposite limb using a modified Mankin scale. The following variables were used to determine scores: surface (0-6), cells (0-4), hypocellularity (0-3), matrix staining (transitional zone [0-4], radiate zone [0-4], and focal empty lacunae or hypereosinophilic cells (0-3). Differences in scores for all response variables were calculated as treated limb minus sham limb. Response variables were formed: score >0 recoded as 1 (favorable response treated better than sham), score of 0 recoded as 2 (neutral response no differences), and score <0 recoded as 3 (unfavorable response treated worse than sham). Bipolar ablative probe-treated limbs had 14.29% favorable responses and 35.71% favorable or neutral responses, whereas shave-treated limbs had 0% favorable and only 7.14% favorable or neutral responses. For all variables, bipolar ablative probe-treated limbs had more favorable responses. The less severe histological change in the bipolar ablative probe-treated joints compared with the shave-treated joints suggests that bipolar ablation of articular cartilage may be a better treatment for chondromalacia than the usual shaving methods of debridement. Further, there were no pathological changes in the subchondral bone. PMID- 9754477 TI - Shortening of the patellar tendon after anterior cruciate ligament reconstruction. AB - In this prospective study, patellar height changes were investigated after anterior cruciate ligament (ACL) reconstruction with a mean follow-up of 22.4 months. A total of 114 patients were included. Fifty-two patients (group A) were treated by multiple suture repair, 27 patients (group B) underwent acute ACL reconstruction, and 35 patients (group C) underwent ACL reconstruction > or =6 weeks after injury with a patellar tendon graft. The patellar vertical height ratios (VHR) were evaluated preoperatively (VHR 1), 6 months postoperatively (VHR 2), and at follow-up (VHR 3). For the studied questions, the following answers were obtained: (1) The change of the patella height was the same in all three groups (i.e., disregarding the different surgical procedures). (2) The time elapsed between injury and ACL reconstruction did not influence the shortening of the patellar tendon. (3) Women showed a more pronounced shortening of the patellar tendon than did men. (4) A significant shortening of the patellar tendon occurred in 30% of our patients, and the process of shortening was finished 6 months postoperatively. (5) Anterior knee pain was present in 27.2% of our patients and occurred significantly more often after patellar tendon graftings. (6) Age had no influence on the changes of the patellar height. PMID- 9754478 TI - Articular and osseous lesions in recent ligament tears: arthroscopic changes compared with magnetic resonance imaging findings. AB - The treatment of ligament injuries of the knee has undergone rapid progress, especially with the improvement of arthroscopic reconstruction of the anterior cruciate ligament (ACL). Since the advent of magnetic resonance imaging (MRI) after knee trauma with ligament injuries, interest has focussed on the clinical significance of concomitant articular and osseous lesions. In 48 of 141 MRIs, different types of these lesions were found; in 38 cases an arthroscopy was performed and 34 times the patients could clinically and radiologically be examined after at least 6 months. Bone bruise was found 26 times, in 16 cases associated with ACL-tears. Eleven patients had subchondral fractures, 7 osteochondral fractures, and in 4 patients, stress fractures were found. They were attributed to various mechanisms of trauma, in different percentages associated with ligament tears and in different dimensions visible or progressive on follow-up MRIs. Obviously some of the different lesions of subchondral and spongeous bone can indicate later degenerative arthritis, so that we find hints for a modification of rehabilitation, e.g., open versus closed kinetic chain or orthosis with relief of single compartments. PMID- 9754479 TI - Comparison of oral ketorolac and hydrocodone for pain relief after anterior cruciate ligament reconstruction. AB - The analgesic effectiveness of ketorolac tromethamine was compared with hydrocodone and acetaminophen for pain from an arthroscopically assisted patellar tendon autograft anterior cruciate ligament reconstruction. There were 125 patients evaluated in a double-blind, randomized, multicenter, and multidose study. A loading dose of parental ketorolac tromethamine was administered and subjects were later given two staged doses of the same "unknown" drug with pain evaluations conducted after each dose. For group 1, dose 1 consisted of ketorolac tromethamine 20 mg orally and dose 2 was ketorolac tromethamine 10 mg. For group 2, both dose 1 and dose 2 consisted of hydrocodone 10 mg plus acetaminophen 1,000 mg orally. Efficacy was evaluated by standard analgesic measures. Subjects treated as outpatients showed lower categorical pain intensity for ketorolac tromethamine than hydrocodone and acetaminophen at 1 hour (P=.03), 2 hours (P=.006), and 3 hours (P=.02); lower summed intensity differences for ketorolac tromethamine than hydrocodone and acetaminophen at 3 hours (P=.014) and 4 hours (P=.019); and better total pain relief for ketorolac tromethamine than hydrocodone and acetaminophen at 3 hours (P=.014) and 4 hours (P=.013). With an effective loading dose administered before the subsequent oral dosage, there was statistically better pain reduction with ketorolac tromethamine than with hydrocodone and acetaminophen. Moreover, ketorolac tromethamine was no more likely to cause digestive complaints than hydrocodone and acetaminophen. No bleeding problems were observed in either group. In the outpatient setting, ketorolac tromethamine controls postoperative pain better than hydrocodone and acetaminophen in the immediate postsurgery period. PMID- 9754480 TI - A prospective study of pain and analgesic use in outpatient endoscopic anterior cruciate ligament reconstruction. AB - A prospective study was undertaken to evaluate the postoperative pain and analgesic profiles of a group of 50 patients undergoing outpatient anterior cruciate ligament (ACL) reconstruction and to compare their profiles with those of a group of 50 patients undergoing outpatient non-ACL arthroscopic surgery. All patients received preoperative and postoperative ketorolac, intraincisional/intra articular bupivacaine, intraoperative ketorolac, and propofol anesthetic. The percentage of patients receiving supplemental analgesia in the recovery room was 49% (average, 2.2 mg intravenous morphine sulfate) for the ACL group and 31% (average, 1.2 mg intravenous morphine sulfate) in the non-ACL group. Narcotic use and pain scores peaked in both groups on postoperative days 1 and 2. The ACL group used significantly more narcotic and had higher pain scores in the first week after surgery than did the non-ACL group. However, there were no subsequent admissions, readmissions, or emergency room visits for pain. All were satisfied with the outpatient nature of this surgery. Patients tolerate outpatient endoscopic ACL reconstruction with moderate pain and narcotic use. Outpatient endoscopic ACL reconstruction can be performed safely, effectively, and with considerable cost savings. PMID- 9754481 TI - The use of arthroscopy to document accurate position of core decompression of the hip. AB - The use of hip arthroscopy is documented as a means of determining accurate placement for core decompression of the femoral head. The authors describe the technique whereby the patient is placed on the fracture table in the supine position and the guide wire for the core decompression is inserted into the middle of the infarct. The surgeon is assured of accurate placement within the center of the infarct. PMID- 9754482 TI - Pathological mediopatellar plica found in the knee of an infant. AB - We report the case of a pathological mediopatellar plica found in the right knee of a 15-month-old infant girl. Flexion contracture of the knee was found to be 40 degrees. An arthroscopic view showed a large and thick voluminous mediopatellar plica. It was trapped between the patella and the medial femoral condyle and it was in tight contact with the medial facet of the patella at 40 degrees flexion in the knee, blocking further extension. A longitudinal groove was noted on the articular surface of the medial femoral condyle that looked as if it had another trochlea on arthroscopic view. The mediopatellar plica came into contact with the groove at 60 degrees flexion in the knee and it fitted precisely into the groove at further flexion. The pathological plica was resected arthroscopically, which resulted in approximately 10 degrees improvement in extension of the knee. Histological examination found hypertrophy and chronic nonspecific inflammation of the synovium. The patient was helped with range-of-motion exercise and quadriceps-strengthening exercise. At 27 months follow-up, the knee had gained full extension. This article reports that a pathological mediopatellar plica may develop in infants. PMID- 9754483 TI - Arthroscopic removal of a .44 caliber bullet from the hip. AB - Hip arthroscopy is far less invasive than standard open arthrotomy and offers unparalleled visualization of the acetabulum and femoral head. Diagnostic arthroscopy is becoming increasingly accepted as therapeutic options are still evolving. We report the case of the arthroscopic removal of a .44 caliber bullet from the femoral head of a 45-year-old man. The procedure afforded the opportunity to thoroughly irrigate the joint, debride the articular surface, and remove several loose bodies. PMID- 9754484 TI - Synovial osteochondromatosis of the cruciate ligament. AB - An unusual case of synovial chondromatosis of the cruciate ligaments is reported that resulted principally in a loss of function, secondary to a mechanical block to extension. Magnetic resonance imaging was useful in directing surgery, but not in making the formal diagnosis. PMID- 9754485 TI - Osteochondritis dissecans of the medial femoral condyle associated with congenital hypoplasia of the lateral meniscus and anterior cruciate ligament. AB - We report a patient with osteochondritis dissecans of the medial femoral condyle associated with congenital hypoplasia of the lateral meniscus and anterior cruciate ligament. This is the first report of such a case. PMID- 9754486 TI - Calcium pyrophosphate dihydrate crystal deposition disease after anterior cruciate ligament reconstruction. AB - We report the case of a 34-year-old woman who presented with calcium pyrophosphate dihydrate (CPPD) crystal deposition disease shortly after anterior cruciate ligament (ACL) reconstruction using a polyester artificial ligament (Leeds-Keio; Neoligaments, Leeds, England). The patient had earlier undergone a medial collateral ligament repair of a sprain to her right knee incurred while skiing. Nine years later, she underwent ACL reconstruction. Seventeen months after ACL reconstruction, calcification was observed on radiographs of the medial and lateral menisci. Based on these calcifications and polarized light microscopic findings of the joint fluid, the diagnosis was made of CPPD crystal deposition. CPPD deposition appeared to have resulted from intra-articular damage incurred during ACL reconstruction as well as prolonged anterior instability. PMID- 9754487 TI - The peel-back mechanism: its role in producing and extending posterior type II SLAP lesions and its effect on SLAP repair rehabilitation. AB - A previously undescribed mechanism of injury for posterior Type II SLAP lesions is described. The primary feature of this mechanism is a torsional peel-back of the posterosuperior labrum. Secure fixation by posterior-superior placement of suture anchors into the posterosuperior corner of the glenoid is essential. The repair must be protected against torsional peel-back forces by avoiding external rotation beyond 0 degrees for 3 weeks. PMID- 9754488 TI - Arthroscopic Lachman test: a new technique using anatomic references. AB - Using familiar anatomic references viewed during a standard arthroscopic evaluation, an arthroscopic Lachman test can help the surgeon identify anterior cruciate ligament deficiency. This test is especially useful in those patients where a false-negative report as a result of guarding, meniscal tears or other factors is suspected. Additionally, this test can be used after anterior cruciate ligament reconstruction to verify that the abnormal anterior laxity has been eliminated. PMID- 9754489 TI - A modified endoscopic technique for posterior cruciate ligament reconstruction using allograft. AB - This article describes a modified endoscopic technique of posterior cruciate ligament reconstruction using bone-patellar tendon-bone or calcaneus tendon allografts. Three portals were used: a parapatellar anteromedial portal, a lateral anterolateral portal, and a proximal posteromedial portal. The tibial tunnel was made through the anterolateral tibial cortex 2 cm lateral to the tibial tuberosity to the posterior flat spot of the tibia 1 cm below the articular margin and just lateral to the midline. By this method, a less acute angle at the turning point of the tibial tunnel and a straighter alignment of the graft in the coronal plane can be obtained. The femoral tunnel was made through the lateral anterolateral portal without incision over the medial femoral condyle to minimize the injury to the vastus medialis obliques muscle. The 25-mm long proximal bone plug was easily passed through the tibial tunnel using a specially designed suture pusher and guided into the femoral tunnel by pulling the leading suture with the knee flexed 30 degrees. Firm graft fixation was achieved with absorbable interference screws or staples. PMID- 9754490 TI - Magnetic resonance imaging assessment of the rotator cuff: is it really accurate? PMID- 9754491 TI - Magnetic resonance imaging (MRI) of the knee and patellofemoral joint. PMID- 9754492 TI - Transsection of the peroneal nerve complicating knee arthroscopy: case report and cadaver study. PMID- 9754493 TI - Recurrence rate using Caspari's technique for arthroscopic Bankart repair. PMID- 9754495 TI - Men and domestic violence. PMID- 9754494 TI - Asking the right questions--physician-assisted suicide and emergency medical systems. PMID- 9754496 TI - Do quantitative changes in pain intensity correlate with pain relief and satisfaction? AB - OBJECTIVE: To correlate measured pain intensity (PI) changes with pain relief and satisfaction with pain management. METHODS: A prospective single-group repeated measures design study. A heterogeneous group of patients were asked to record their levels of PI at initial presentation and at ED release using a numerical descriptor scale (NDS) and a visual analog scale (VAS). At release, a 5-point pain relief scale and a pain management satisfaction survey were also completed. RESULTS: A convenience sample of 81 patients were enrolled over the study period. The average reduction in PI for all patients was 33%. A 5%, 30%, and 57% reduction in PI correlated with "no," "some/partial," and "significant/complete" relief, respectively (p < 0.001). However, when patients were divided into 2 groups based on their initial PI scores, patients with moderate/severe pain (NDS > 5) required a reduction of 35% and 84% in PI to achieve "some/partial" and "significant/complete" relief, respectively. Patients in less pain (NDS < or = 5) needed 25% and 29% reductions in PI for the same categories (p=0.8). Patients were generally satisfied with their pain management. There was a positive association between pain relief and satisfaction with pain management. CONCLUSION: There is a significant association between changes in PI and pain relief. Greater reductions in PI are required for patients presenting with more severe initial pain to achieve relief compared with those who have lesser initial PI. While there is a linear relationship between increasing pain relief and satisfaction, relief of pain appears to only partially contribute to overall satisfaction with pain management. PMID- 9754497 TI - Pulse oximetry as a fifth vital sign in emergency geriatric assessment. AB - OBJECTIVE: To determine the utility of pulse oximetry as a routine fifth vital sign in emergency geriatric assessment. METHODS: Prospective study using pulse oximetry to measure O2 saturation in geriatric patients presenting to ED triage. Saturation values were disclosed to clinicians only after they had completed medical evaluations and were ready to release or admit each patient. The authors measured changes in medical management and diagnoses initiated after the disclosure of pulse oximetry values. The study included 1,963 consecutive adults aged > or = 65 years presenting to triage at a university ED. Measurements included changes in select diagnostic tests: chest radiography, complete blood count (CBC), spirometry, arterial blood gases (ABGs), pulse oximetry, and ventilation-perfusion scans; treatments: antibiotics, beta-agonists, and supplemental O2; and hospital admission and final diagnoses that occurred after complete ED evaluation when physicians were informed of triage pulse oximetry values. RESULTS: 397 (20.2%) geriatric patients had triage pulse oximetry values <95%. Physicians ordered repeat oximetry for 51 patients, additional chest radiography for 23, CBC for 16, ABGs for 15, spirometry for 5, and ventilation perfusion scans for none. Physicians ordered 49 new therapies for 44 patients, including antibiotics for 14, supplemental O2 for 29, and beta-agonists for 6. Nine patients initially scheduled for ED release were subsequently admitted to the hospital. Physicians changed or added diagnoses for 27 patients. CONCLUSIONS: Using pulse oximetry as a routine fifth vital sign resulted in important changes in the diagnoses and treatments of a small proportion of emergency geriatric patients. PMID- 9754498 TI - Male victims of domestic violence and their history of perpetrating violence. AB - OBJECTIVE: To determine whether male victims of domestic violence have similar rates of violence perpetration compared with men evaluated in the ED with other causes of injury. METHODS: Case-control retrospective ED record review with linkage to police department records. Cases were identified by ICD code N-code 995.81 (adult maltreatment syndrome) over a 4-year period (January 1, 1991, to December 31, 1994) at one urban trauma center. Medical records were reviewed to confirm that the assailant was an intimate female partner. Controls were identified by E-codes 880-888 (unintentional falls) and matched by age, race, and date of visit. All names were linked to police department record information regarding arrests for domestic violence perpetration, nonaggravated assaults, aggravated assaults, firearms violations, and driving under the influence of alcohol (DUI). This information was reported without patient identifiers. Comparisons between cases and controls were made with chi2 analysis. RESULTS: Forty-five cases and 45 controls were identified. The cases were injured by unarmed fights, E960 (31%); cuttings, E966 (33%); blunt objects, E968.2 (31%); and bites, E968.8 (5%). Median age (interquartile range) for cases was 32 (25.75, 38.25) years and for controls was 31 (25, 36.5) years. Median follow-up (interquartile range) of police records after ED visit was 45 (37, 50) months for cases and 45 (36.75, 51) months for controls. Fifty-one percent of the cases had arrests for domestic violence perpetration vs 22.2% of the controls (p=0.009). Forty-four percent of the cases had been arrested for nonaggravated assaults vs 20.0% of the controls (p=0.024). There was no statistical difference between the cases and controls in arrests for aggravated assaults (13.3% vs 4.4%), firearm violations (22.2% vs 17.8%), or DUIs (35.6% vs 20%). CONCLUSION: Men who present to the ED with injuries inflicted by their female partners have a high rate of domestic violence perpetration. This information calls into question whether many male "victims" of domestic violence are injured in self-defense by the female "victim." Also, injury by a female partner may be a useful indicator to identify batterers, so they can be referred by appropriate resources. PMID- 9754499 TI - Ambulatory blood pressure and Holter monitoring of emergency physicians before, during, and after a night shift. AB - BACKGROUND: Occupational stress may affect measured hemodynamic and electrocardiographic variables. Data describing the physiologic effects of work on the emergency physician (EP) are sparse. OBJECTIVE: To determine whether blood pressure (BP) and heart rate variability (HRV) of the EP are affected during a night shift in the ED. METHODS: This prospective study evaluated BP and HRV in attending EPs at an urban academic medical center for a 24-hour period during which a night shift was scheduled. Participants were fitted with an oscillometric ambulatory BP device and a Holter monitor at 1500 hours on the day of a night shift. The monitors were worn continuously before, during, and after a night shift (2300-0700) in the ED and were removed at 1500. Systolic BP (SBP), diastolic BP (DBP), mean arterial pressure (MAP), heart rate (HR), measures of HRV, and occurrence of cardiac dysrhythmias were evaluated. Comparisons were made for ED and non-ED awake periods and non-ED sleep periods. RESULTS: Twelve participants completed the study. Eight (67%) subjects were men and 4 (33%) were women. Age ranged from 28 to 40 years (mean 34.1+/-4.1). Results were analyzed using repeated-measures ANOVA. An elevation of mean DBP (5.5 mm Hg+/-4.37; p < 0.05; 95% CI 1-10) during night shift activity was seen. A trend toward elevation of SBP, MAP, and HR was discernible. HRV measures indicated a significant relative increase in sympathetic vs parasympathetic tone and an increase in HR of prework and work compared with postwork. Dysrhythmias observed included sinus tachycardia, sinus bradycardia, sinus pause, atrial premature beats, atrial couplets and triplets, supraventricular tachycardia, and premature ventricular contractions. CONCLUSIONS: The elevation of DBP during a night shift suggests that these patterns of BP variability are activity- or stress-related rather than a result of a true diurnal variation. HRV analysis suggests that sympathetic tone is heightened both before work and during work. The implications of such findings to the health of the EP warrant further investigation. PMID- 9754500 TI - Out-of-hospital intravenous access: unnecessary procedures and excessive cost. AB - OBJECTIVE: To evaluate the concordance with criteria developed by the study investigators and supply costs associated with placement of i.v. lines and saline locks by paramedics in the out-of-hospital setting. METHODS: This was a retrospective consecutive case series at an urban base hospital. Patients were treated by paramedics using one base hospital for medical control during December 1995. Base hospital written records and taped patient calls were reviewed to determine actual i.v. access method used by paramedics, chief complaint, and whether fluid administration was ordered. Indicated method of i.v. access was determined for each patient based on predetermined criteria developed by the investigators. i.v. access methods were ranked by cost of supplies as follows: i.v. line (i.v.) > saline lock (SL) > no i.v. line (No i.v.). An assignment of concordant treatment was made when actual = indicated method, discordant overtreatment when actual > indicated, and discordant-undertreatment when actual < indicated. RESULTS: 452 patients were treated via radio by the base hospital during the study period. 380 of 452 (84%) received an i.v.. 28 of 380 (7%) received fluid resuscitation in the field. 166 of 452 (37%) received concordant treatment; 253 (56%) discordant-overtreatment; and 33 (7%) discordant undertreatment. Pediatric patients (< or =14 years of age) were more likely to be undertreated as compared with adults, 33% vs 3% (p < 0.001). Patients who had medical chief complaints were more likely to receive discordant-overtreatment as compared with patients who had trauma chief complaints, 61% vs 32% (p < 0.001). 73% of chest pain patients received discordant-overtreatment. Based on these data, the yearly cost of supplies used in i.v. access discordant-overtreatment was $13,735 for this base hospital and $560,000 for the Los Angeles County emergency medical services (EMS) system. 91% of the excess supply cost is due to patients' receiving an i.v. instead of a SL. CONCLUSION: Based on study criteria for utilization of i.v. lines vs SLs in the field, paramedics and base hospital personnel often provide discordant-overtreatment of patients by placement of an i.v. when a SL or No i.v. would suffice, resulting in unnecessary costs for EMS systems. PMID- 9754501 TI - Screening for geriatric problems in the emergency department: reliability and validity. Identification of Seniors at Risk (ISAR) Steering Committee. AB - OBJECTIVE: To determine the test-retest reliability and concurrent criterion validity of a self-report ED screening questionnaire for adverse outcomes in elders. METHODS: A cohort of 1,885 patients aged > or = 65 years were recruited from the EDs of 4 Montreal hospitals. Patients were excluded if they could not be interviewed because of their clinical status or cognitive impairment and no informant was available. The screening questionnaire, administered in the ED, contained 27 items on social, physical, and mental risk factors, medical history, and use of hospital services, medications, and alcohol. A random sample of 404 patients were invited to participate in a clinical assessment 1-3 weeks after the ED visit, that included re-administration of the screening questionnaire, and standardized instruments to assess disability, social resources, depression, alcohol use and abuse, and current medications. RESULTS: Study data were collected from 221 patients (54.7%), of whom 193 were included in the test-retest reliability analyses and 213 in the analyses of concurrent validity. The concordance correlation coefficient for test-retest reliability of the risk factor score was 0.78 (95% confidence interval: 0.71, 0.83; n=193). Several screening questions showed moderately good agreement with the appropriate criterion standard, particularly those on visual and hearing impairment, depression, and use of medications. The best subset of 9 screening questions explained approximately half of the variance in the total disability score. CONCLUSIONS: The screening questionnaire score has good test-retest reliability, but individual screening questions have, at best, modest concurrent validity. The final set of screening questions should be selected based on their predictive validity. PMID- 9754502 TI - Intraarterial vs intravenous administration of antivenin for the treatment of Crotalidae atrox envenomation: a pilot study. AB - OBJECTIVE: Standard therapy for significant snake envenomation includes antivenin. i.v. administration is currently the only recommended route. Intraarterial (i.a.) administration has potential advantages over i.v. that could improve outcome. To study this, the authors compared i.v. and i.a. antivenin administrations for the treatment of experimental snake envenomations. METHODS: 14 adult female swine were anesthetized and prepared with femoral artery and ear vein catheters, and baseline hoof, forearm, and thigh circumference and volume displacement measurements were taken. Crotalidae atrox venom was injected into the subcutaneous tissue of the hoof. The doses of venom were 4.75, 9.50, 19.00, 37.90, 47.30, 56.90, and 66.40 mg. Immediately following injection of venom, polyvalent antivenin (Crotalidae) (0.285 mg/10 mL saline) was infused over 30 minutes into the femoral artery (i.a. group) or ear vein (i.v. group). As a control, 10 mL of saline was infused into the ear vein (i.a. group) or femoral artery (i.v. group). Measurements were recorded up to 48 hours. Linear mixed effect regression models were used for each measurement and to compare the i.a. and i.v. groups. RESULTS: Venom dose and time after administrations were associated with increased circumferences and increased volumes (p < 0.05). i.v. administration was associated with larger hoof (1.26 cm) and forearm (0.42 cm) sizes and volume displacement (21.71 mL) when compared with i.a. administration ( p < 0.05). CONCLUSION: i.a. antivenin results in a modest but significant decrease in tissue edema when compared with i.v.. PMID- 9754503 TI - Emergency management of migraine: is the headache really over? AB - OBJECTIVES: To observe headache frequency after release for acute migraine sufferers treated in an ED; to observe the impact after-release headaches and associated symptoms have on quality of life; and to document the variability in migraine management in an emergency setting. METHODS: Prospective observational study, including 24- and 72-hour telephone follow-up. RESULTS: Over a 4-month period, 143 patients with headaches (149 visits) were observed in the ED. Of 108 patients successfully contacted, the incidence of headache in the first 24 hours after release was 49.1%. Forty-two patients left the ED without pain; 13 of these subsequently had return of headache. Sixty-six left with some degree of pain, with 40 having headache persistence at 24 hours. The difference in 24-hour headache rate between the 2 groups is significant (p=0.008). Five patients still had headaches at 72 hours after release, but 54 of 70 contacted had taken medication for their symptoms between 24 and 72 hours after release. Forty-five percent were not back to normal function at 24 hours, while 21 of 70 were still not sleeping well at 72 hours. Finally, 8 different classes of medications were used in the ED for migraine headaches, with 20 patients receiving at least 3 types of medication. CONCLUSIONS: Treatment for acute migraine headache in this emergency setting was variable. Patients not obtaining complete relief in the ED had a higher rate of headache after release than did those who left with no pain. Migraine sufferers may have normal daily function affected for up to 72 hours or longer after ED treatment. PMID- 9754504 TI - New-onset generalized seizures in patients with AIDS presenting to an emergency department. AB - OBJECTIVE: To determine the etiology of new-onset generalized seizures in patients with AIDS presenting to an ED. Patients without HIV infection with a first-time seizure were used as a comparison group. With these data, the current American College of Emergency Physicians (ACEP) guidelines on the workup of new onset seizures were applied to determine whether they could safely be used in patients with AIDS. METHODS: The authors conducted a retrospective review of all patients with new-onset generalized seizures who presented to an academic medical center hospital ED in New York City over 2 years. A standard ED medical evaluation with history, physical examination, and routine laboratory studies including at least a panel 7 chemistry, serum magnesium, and complete blood count was performed. RESULTS: The causes of new-onset seizures in 26 patients with AIDS were idiopathic (8), HIV encephalopathy (8), CNS toxoplasmosis (5), alcohol withdrawal (2), progressive multifocal leukoencephalopathy (2), and CNS lymphoma (1). In 120 patients without HIV infection, idiopathic (43) and alcohol withdrawal (29) were the most common diagnoses. Six patients with AIDS had CNS lesions necessitating immediate admission to the hospital (5 with toxoplasmosis and 1 with lymphoma). Only 2 of 6 had findings on initial ED examination that would have suggested admission under current guidelines written for patients without HIV infection. The 4 patients with no findings were 3 with CNS toxoplasmosis and 1 with CNS lymphoma. CONCLUSION: Four of 26 AIDS patients with immediately treatable CNS lesions could have been sent home for outpatient evaluation of their seizures on the basis of current guidelines for non-HIV infected patients. However, the updated 1997 ACEP guidelines now include emergent brain neuroimaging studies on patients who have or are suspected of having AIDS. This study helps to strengthen this recommendation. Based on these findings, the authors suggest a neuroimaging study with a lumbar puncture, if indicated, in the ED or inpatient admission workup for all patients with AIDS or suspected AIDS presenting with new-onset generalized seizures. PMID- 9754505 TI - Oregon emergency medical technicians' attitudes toward physician-assisted suicide. AB - OBJECTIVES: To determine Oregon intermediate and advanced emergency medical technicians' (EMTs') attitudes toward physician-assisted suicide (PAS) and factors associated with those attitudes. METHODS: An anonymous survey was sent to a random sample of 498 EMTs registered in Oregon. RESULTS: Surveys were delivered to 498 EMTs and 343 completed surveys were returned, for a response rate of 69%. The mean age of the respondents was 37.5 years (+/-8.73) and 79% were male. 232 (68%) agreed that PAS should be legal, 263 (77%) agreed that terminally ill people have a right to decide to commit suicide, while 57 (17%) thought not attempting resuscitation would be immoral. 251 (73%) reported seeing attempted suicide in terminally ill patients at least once, with 117 (34%) experiencing such calls >5 times. Only 22 (6%) stated that they would be unable to work in a system that directed them to withhold resuscitation after a PAS attempt, and 277 (81%) agreed that treatment protocols should direct EMTs to withhold resuscitation. 105 (31%) thought EMTs should participate in the decision to withhold resuscitation. 206 (60%) thought the law should allow lethal injection for terminally ill patients. 201 (59%) agreed there were circumstances under which they might personally consider PAS. If PAS were legal, EMTs stated they would withhold treatment from a terminally ill patient following attempted suicide in the following circumstances: based on standing orders, 78%; with on line medical direction, 67%; after speaking with the primary physician, 53%; if the patient had decision-making capacity, 45%; with written documentation from the patient's physician, 68%; and never, 6%. CONCLUSIONS: A majority of Oregon EMTs responding to this survey expressed support for PAS, think treatment protocols should direct paramedics to withhold resuscitation in such cases, and would feel comfortable withholding resuscitation given appropriate protocols. Nearly 3 out of 4 Oregon EMTs report seeing at least 1 terminally ill patient who had attempted suicide. PMID- 9754506 TI - Assessment of airway visualization: validation of the percentage of glottic opening (POGO) scale. AB - OBJECTIVE: Research defining optimal methods of intubation has been limited by the lack of a validated outcome measure to assess airway visualization. The objective of this study was to develop a reliable scale for the assessment of airway visualization during endotracheal intubation. METHODS: This prospective study was performed to assess the intra- and interphysician reliabilities of emergency physicians (EPs) for estimating the percentage of glottic opening (POGO) that is visualized during direct laryngoscopy. Using video images of laryngeal views obtained from a commercially available videotape, still slide images were prepared representing glottic openings ranging from 0% to 100%. Five EPs, blinded to study objective, reviewed 25 pairs of airway slides (50 slides total). For each slide, the physicians recorded the POGO and their scores using a modified Cormack-Lehane (MCL) scale, where grade I is a view of the full glottic opening, MCL grade II is a partial view of the glottic opening, and MCL grade III is a view of the epiglottis only. Inter- and intraphysician reliabilities were assessed using the kappa statistic (K) for MCL grade and intraclass correlation coefficient for the POGO scores. RESULTS: For the POGO score, the degree of intrarater reliability was very good, with an intraphysician correlation of 0.85 and an interphysician correlation of 0.74. For the MCL score, the intraphysician concordance had a K of 0.71, and interphysician concordance was also good, with a kappa of 0.59. CONCLUSION: Both the modified version of the Cormack-Lehane grading classification and the POGO score have good interphysician and intraphysician reliabilities. Because the POGO score can distinguish patients with large and small degrees of partial glottic visibility, it might provide a better outcome for assessing the difference between various intubation techniques. PMID- 9754507 TI - Research fundamentals: I. Getting from hypothesis to manuscript: an overview of the skills required for success in research. AB - This is the first in a series of articles developed by members of the Society for Academic Emergency Medicine (SAEM) Research Committee. The purpose of this series is to describe a stepwise approach to research, from the inception of a hypothesis to the final publication of a report. This series is written for junior academic emergency physicians (EPs), as well as nonacademic physicians who have an interest in research. This first article presents an overview of the steps involved in performing research and publishing the results, emphasizing the initial steps and the importance of collaboration. PMID- 9754508 TI - Statistical methodology: VI. Mathematical modeling of the electrocardiogram using factor analysis. AB - The ECG is a 12-lead-vector system and is known to contain redundant information. Factor analysis (FA) is a statistical technique that improves measured data and eliminates redundancy by identifying a minimum number of factors accounting for variance in the data set. OBJECTIVE: To identify the minimum number of lead vectors required to predict the 12-lead ECG. METHODS: A total of 104 ECGs were obtained from 24 normal men, 22 normal women, and 28 men and 30 women with variable pathologies. Each ECG lead was simultaneously acquired and digitized, resulting in a voltage-time data array stored for mathematical analysis. Each array was factor-analyzed to identify the minimum number of lead-vectors spanning the ECG data space. The 12-lead ECG was then predicted from this minimum lead vector set. ANOVA was used to test for statistical significance between normal and pathologic data groups. RESULTS: FA revealed that 3 lead-vectors accounted for 99.12%+/-0.92% (95% CI+/-0.18%) of the variance contained in the 12-lead ECG voltage-time data for all 104 cases. There were no statistically significant differences between men and women (99.25%+/-0.66% vs 98.98+/-1.11%; p=0.139). Statistically significant differences were noted between normal and acute myocardial infarction ECGs (99.5%+/-0.27% vs 98.66+/-1.25%; p=0.00003). The measured and predicted leads were almost identical. A 3-dimensional spatial ECG derived from the 3-lead-vector set resulted in variable curved surfaces that differed by pathology. CONCLUSIONS: The 12-lead ECG can be derived from only 3 measured leads and graphed as a 3-D spatial ECG. This type of data processing may lead to instantaneous acquisition and may enhance the diagnostic capability of the ECG from routine bedside telemetry equipment. PMID- 9754509 TI - Emergency department initiatives to improve the public health. PMID- 9754510 TI - Emergency medications via the endotracheal tube: when is this route preferred? PMID- 9754511 TI - Inhaled budesonide: placebo problems. PMID- 9754512 TI - Outcomes of telephone medical care. AB - OBJECTIVES: To document the outcomes of a telephone coverage system and identify patient characteristics that may predict these outcomes. DESIGN: Telephone survey. SETTING: An academic outpatient medical practice that has a physician telephone coverage service. PATIENTS: All patients (483) who called during the 3 week study period to speak to a physician were evaluated, and for the 180 patients with symptoms, attempts were made to survey them by telephone 1 week after their initial telephone call. MEASUREMENTS AND MAIN RESULTS: The mean age of the 180 patients was 41 years, 71% were female, and 56% belonged to commercial managed care plans. In the week after the initial telephone call, the following outcomes were reported: 27% of the patients had no further contact with the practice; 9% filled a prescription medication; 19% called the practice again; 48% kept an earlier appointment in the practice; 3% saw an internist elsewhere; 8% saw a specialist; 8% went to an emergency department; 4% were admitted to a hospital. Of the 180 patients who called with symptoms, 160 (89%) were successfully contacted for survey. Eighty-seven percent of these 160 patients rated their satisfaction with the care they received over the telephone as excellent, very good, or good. In multivariate analysis, patients' own health perception identified those most likely to have symptom relief (p = .002), and symptom relief, in turn, was a strong predictor of high patient satisfaction (p = .006). Thirty-three percent of the 160 patients reported that they would have gone to an emergency department if a physician were not available by telephone. CONCLUSIONS: In the present study, younger patients, female patients, and patients in commercial managed care plans used the telephone most frequently. Also, the telephone provided a viable alternative to emergency department and walk-in visits. Overall satisfaction with telephone medicine was high, and the strongest predictors of high patient satisfaction were symptom relief and patients' own health perception. PMID- 9754513 TI - HIV/AIDS patients' perspectives on adhering to regimens containing protease inhibitors. AB - OBJECTIVE: To gather qualitative data regarding HIV/AIDS patients' perspectives about HIV-1 protease inhibitors (PIs), and about their experiences taking and adhering to regimens containing PIs. DESIGN: Six focus groups of persons under care for HIV were conducted between September and November 1996 regarding participants' knowledge, awareness, experiences when taking, and adherence to antiretroviral regimens containing PIs. An identical discussion guide was used to facilitate all six groups. Focus group proceedings were audiotaped, transcribed, coded for themes, and analyzed qualitatively. SETTING: HIV/AIDS practices of three teaching hospitals and two community health centers. PATIENTS/PARTICIPANTS: Fifty-six patients with HIV disease: 28 men and 28 women. MEASUREMENTS AND MAIN RESULTS: Knowledge and positive impressions of PIs were prevalent among this diverse group of persons with HIV, and did not differ by race/ethnicity or gender. Most knew that these were new, potent medications for treating HIV/AIDS. Networks of persons with HIV and medical providers were the most important information sources. Those taking PIs were aware that adherence to the regimen is important, and most were using special strategies to maximize their own adherence, but expressed considerable frustration about the central role these medication regimens had assumed in their life. A subset who did not believe they would adhere to these regimens had declined treatment with them. Motivating factors for taking and adhering to these complex regimens were improving CD4 counts and viral loads and the patient-provider relationship. CONCLUSIONS: Among those with HIV/AIDS, awareness of PIs and their effectiveness is substantial, owing to the impact of informal networks and medical providers. This early positive "reputation" of PIs may enhance motivation for adherence. Those who are taking PIs invest substantial effort adhering to these complex regimens, but resent the need to make medications the focus of their lives. PMID- 9754514 TI - Effect of histamine-2 receptor antagonists on blood alcohol levels: a meta analysis. AB - OBJECTIVE: To determine the effect, if any, of histamine type 2 receptor antagonists (H2RAs) on serum alcohol levels under various conditions including type of H2RA receptor antagonist, alcohol dose, and fed status of the subject. STUDY DESIGN: Meta-analysis of the published literature. DATA SOURCES: Studies were identified by MEDLINE (January 1982 through December 1997) using the key words H2 receptor antagonists and alcohol. Other studies were identified by reviewing bibliographies of retrieved articles and by discussion with colleagues. STUDY SELECTION: Studies were selected if they involved the coadministration of H2RAs and alcohol in healthy, human volunteers. Studies that may have addressed this goal but were performed in another context, for instance the measurement of ulcer healing, were excluded. DATA EXTRACTION: Data were extracted on the design, number of participants, participant characteristics, type and dose of H2RA administered, serum alcohol levels (measured as Cmax) along with standard deviations, dose of alcohol received, and fed or fasted status of participants. Alcohol dose was arbitrarily divided into low dose (< or = 0.5 g/kg body weight) versus high dose (> 0.5 g/kg body weight). In addition, studies involving ranitidine and cimetidine were stratified by sample size into small (n < or = 10) versus not small (n > 10). MEASUREMENTS AND MAIN RESULTS: Twenty-four trials met selection criteria. Small elevations in Cmax were noted when cimetidine (2.71 mg/DL; 95% confidence internal [CI] 1.60, 3.83) or ranitidine (6.95 mg/DL; 95% CI 5.83, 8.08) were coadministered with alcohol. No such differences were noted for famotidine (0.28 mg/DL; 95% CI -1.24, 1.80) or nizatidine (2.33 mg/DL;, 95% CI 0.06, 4.72). The elevation detected with cimetidine and ranitidine was most pronounced in smaller studies (n < 10). Separate analyses investigating the effect of alcohol dose and fed or fasted status of participants revealed no clinically important differences. CONCLUSIONS: Cimetidine and ranitidine, but not the other H2RAs, can cause small elevations of serum alcohol level when alcohol and drug are administered concurrently. Studies with larger numbers of participants were less likely to demonstrate this effect. Relative to accepted, legal definitions of intoxication, the effect of any H2RA on blood alcohol level is unlikely to be clinically relevant. PMID- 9754515 TI - Quinine for nocturnal leg cramps: a meta-analysis including unpublished data. AB - OBJECTIVE: With respect to the use of quinine for the treatment of nocturnal leg cramps, to determine whether the findings of a previously performed meta-analysis of published data are altered with the addition of unpublished data, and whether publication bias is present in this area. DESIGN: A meta-analysis of eight (four published and four unpublished) randomized, double-blind, placebo-controlled trials, seven of which had a crossover design. SETTING: Randomized trials that were available as of July 1997. SUBJECTS: Ambulatory patients (659) who suffered from regular nocturnal leg cramps. MAIN RESULTS: When individual patient data from all crossover studies were pooled, persons had 3.60 (95% confidence interval [CI] 2.15, 5.05) fewer cramps in a 4-week period when taking quinine compared with placebo. This compared with an estimate of 8.83 fewer cramps (95% CI 4.16, 13.49) from pooling published studies alone. The corresponding relative risk reductions were 21% (95% CI 12%, 30%) and 43% (95% CI 21%, 65%), respectively. Compared with placebo, the use of quinine was associated with an increased incidence of side effects, particularly tinnitus. Publication bias is present in the reporting of the efficacy of quinine for this indication, as almost all published studies reported larger estimates of its efficacy than did unpublished studies. CONCLUSIONS: This study confirms that quinine is efficacious in the prevention of nocturnal leg cramps. However, its benefit may not be as large as reported from the pooling of published studies alone. Given the side effect profile of quinine, nonpharmacologic therapy (e.g., regular passive stretching of the affected muscle) is the best first-line treatment. For persons who find this ineffective and whose quality of life is significantly affected, a trial of quinine is warranted. Prescribing physicians must closely monitor the risks and benefits in individual patients. Publication bias is present in this area even though there is controversy about the role of quinine in the treatment of leg cramps. To minimize the possibility of this bias, persons performing medication related meta-analyses should seek high-quality unpublished data from drug regulatory agencies and pharmaceutical companies. PMID- 9754516 TI - Physical abuse among depressed women. AB - OBJECTIVE: To provide estimates of physical abuse and use of health services among depressed women in order to inform efforts to increase detection and treatment of physical abuse. DESIGN: Retrospective assessment of abuse and health services use over 1 year in a cohort of depressed women. SETTING: Statewide community sample from Arkansas. PARTICIPANTS: We recruited 303 depressed women through random-digit-dial screening. MEASUREMENTS AND MAIN RESULTS: Exposure to physical abuse based on the Conflict Tactics Scale, multi-informant estimate of health and mental health services. Over half of the depressed women (55.2%) reported experiencing physical abuse as adults, with 14.5% reporting abuse during the study year. Women abused as adults had significantly more severe depressive symptoms, more psychiatric comorbidity, and more physical illnesses than nonabused women. After controlling for sociodemographic and severity-of-illness factors, recently abused, depressed women were much less likely to receive outpatient care for mental health problems as compared to other depressed women (odds ratio [OR] 0.3; p = .013), though they were more likely to receive health care for physical problems (OR 5.7, p = .021). CONCLUSIONS: Because nearly all depressed women experiencing abuse sought general medical rather than mental health care during the year of the study, primary care screening for physical abuse appears to be a critical link to professional help for abused, depressed women. Research is needed to inform primary care guidelines about methods for detecting abuse in depressed women. PMID- 9754518 TI - Diagnosing delirium by telephone. AB - To determine whether delirium can be diagnosed by telephone, we interviewed 41 subjects aged 65 years or older 1 month after repair of hip fracture, first by telephone and then face-to-face. Interviews included the modified telephone Mini Mental State Examination and the Delirium Symptom Interview. Delirium was diagnosed using the Confusion Assessment Method diagnostic algorithm, and the telephone results were compared with the face-to-face results (the "gold standard"). Of 41 subjects, 6 were delirious by face-to-face assessment; all 6 were delirious by telephone (sensitivity 1.00). Of 35 patients not delirious by face-to-face assessment, 33 patients were not delirious by telephone (specificity = 0.94). We conclude that telephone interviews can effectively rule out delirium, but the positive diagnosis should be confirmed by a face-to-face assessment, especially in populations with a low prevalence of delirium. PMID- 9754517 TI - Ambulatory health care use by patients in a public hospital emergency department. AB - OBJECTIVE: To describe primary care clinic use and emergency department (ED) use for a cohort of public hospital patients seen in the ED, identify predictors of frequent ED use, and ascertain the clinical diagnoses of those with high rates of ED use. DESIGN: Cohort observational study. SETTING: A public hospital in Atlanta, Georgia. PATIENTS: Random sample of 351 adults initially surveyed in the ED in May 1992 and followed for 2 years. MEASUREMENTS AND MAIN RESULTS: Of the 351 patients from the initial survey, 319 (91%) had at least one ambulatory visit in the public hospital system during the following 2 years and one third of the cohort was hospitalized. The median number of subsequent ED visits was 2 (mean 6.4), while the median number of visits to a primary care appointment clinic was O (mean 1.1) with only 90 (26%) of the patients having any primary care clinic visits. The 58 patients (16.6%) who had more than 10 subsequent ED visits accounted for 65.6% of all subsequent ED visits. Overall, patients received 55% of their subsequent ambulatory care in the ED, with only 7.5% in a primary care clinic. In multivariate regression, only access to a telephone (odds ratio [OR] 0.48; 95% confidence interval [CI] 0.39, 0.60), hospital admission (OR 5.90; 95% CI 4.01, 8.76), and primary care visits (OR 1.68; 95% CI 1.34, 2.12) were associated with higher ED visit rates. Regular source of care, insurance coverage, and health status were not associated with ED use. From clinical record review, 74.1% of those with high rates of use had multiple chronic medical conditions, or a chronic medical condition complicated by a psychiatric diagnosis, or substance abuse. CONCLUSIONS: All subgroups of patients in this study relied heavily on the ED for ambulatory care, and high ED use was positively correlated with appointment clinic visits and inpatient hospitalization rates, suggesting that high resource utilization was related to a higher burden of illness among those patients. The prevalence of chronic medical conditions and substance abuse among these most frequent emergency department users points to a need for comprehensive primary care. Multidisciplinary case management strategies to identify frequent ED users and facilitate their use of alternative care sites will be particularly important as managed care strategies are applied to indigent populations who have traditionally received care in public hospital EDs. PMID- 9754519 TI - Women's preferences for specialists who provide cancer screening and general medical care. AB - In order to determine what types of specialists women prefer for medical care, we examined responses from a cross-sectional survey of adult female patients in a health plan of the independent practice association model in the Minneapolis-St. Paul metropolitan area (n = 1,204). The response rate for the survey was 90%. The women expressing a preference (60% of responders) overwhelmingly preferred to see obstetrician-gynecologists for their breast examinations and Pap smears and strongly preferred family physicians or internists for the remainder of their cancer screening and general medical care. Thus, the majority of women expressed preferences for physicians of different specialties to provide their medical care. PMID- 9754521 TI - Management of breast fibroadenomas. AB - OBJECTIVE: To identify from the literature and clinical experience a rational approach to management of fibroadenomas of the breast. METHOD: Recent literature on detection, diagnosis, and natural history of fibroadenomas was reviewed. Experience with over 4,000 women evaluated in the breast clinic at the Tel-Aviv Medical Center contributed to the management strategies suggested by review of the literature. RESULTS: Fibroadenomas of the breast are common, accounting for 50% of all breast biopsies performed. Physical examination, sonography, and fine needle aspiration are effective in distinguishing fibroadenomas from breast cancer. Transformation from fibroadenoma to cancer is rare; regression or resolution is frequent, supporting conservative approaches to follow-up and management. CONCLUSION: Age-based algorithms that allow for conservative management and that limit excision to patients whose fibroadenomas fail to regress are presented. PMID- 9754522 TI - Directory assistance for telephone care: a toll-free way to improve the quality of communication between patients, providers, and investigators. PMID- 9754523 TI - Discussing pharmaceutical gifts. PMID- 9754524 TI - New directions in comparative physiology and biochemistry: mechanisms, adaptations, and evolution. AB - Historically, the discipline of comparative physiology and biochemistry has had two major goals: (1) elucidation of mechanisms and their adaptative significance, and (2) understanding of the evolution of mechanisms and adaptations. In general, the first goal has dominated the field. In a mechanistic/adaptational approach, the diversity of organisms is an experimental parameter in the investigation. Lineage-specific characteristics reveal both how physiological systems work and how different kinds of animals are adapted to different kinds of environments. We believe that this approach is far from outdated, in part because many animal groups have been investigated superficially if at all, and in part because the incorporation of fundamentally new technologies into our discipline permits us to address previously intractable questions about even intensively studied animal groups. In evolutionary physiology and biochemistry, the diversity of lineage specific physiological systems and how they came to be is the subject of investigation. Early attempts to employ the evolutionary approach were not only few in number, they were unsatisfying in outcome because neither phylogenetic nor mechanistic/adaptational knowledge was adequate to serve as a firm foundation. We agree with earlier authors that new and more sophisticated applications of this approach, together with progress in understanding both animal phylogeny and mechanisms/adaptations, all promise to allow us at last to fulfill our second historic goal. In our view, an integration of the two approaches seems to present the most productive trajectory into the next century. PMID- 9754525 TI - Protein catabolism and renal function in lactating northern elephant seals. AB - Northern elephant seals, Mirounga angustirostris, fast completely from food and water during lactation. Previous investigations of maternal investment suggested physiological constraints on the level of energy expenditure during lactation. In this study, two components of phocid fasting physiology, protein sparing and reduced glomerular filtration rate, were examined for effects of changing body composition and lactation duration. Protein catabolism was estimated from 14C urea turnover in five mid- and five late-lactation females. Body composition was determined by using an ultrasound scanner to measure blubber depth coupled with morphometrics. Glomerular filtration rate was measured in five females at mid- and late-lactation using plasma clearance of 3H-inulin. Protein catabolism increased significantly between measurements. The contribution of protein to metabolism varied with body composition and lactation duration. Mass-proportional glomerular filtration rate increased significantly between measurements. These data suggest that conflicting metabolic demands of lactation and fasting might constrain the duration and magnitude of maternal investment in northern elephant seals. PMID- 9754520 TI - Medical risks for women who drink alcohol. AB - OBJECTIVE: To summarize for clinicians recent epidemiologic evidence regarding medical risks of alcohol use for women. METHODS: MEDLINE and PsychINFO, 1990 through 1996, were searched using key words "women" or "woman," and "alcohol." MEDLINE was also searched for other specific topics and authors from 1980 through 1996. Data were extracted and reviewed regarding levels of alcohol consumption associated with mortality, cardiovascular disease, alcohol-related liver disease, injury, osteoporosis, neurologic symptoms, psychiatric comorbidity, fetal alcohol syndrome, spontaneous abortion, infertility, menstrual symptoms, breast cancer, and gynecologic malignancies. Gender-specific data from cohort studies of general population or large clinical samples are primarily reviewed. MAIN RESULTS: Women develop many alcohol-related medical problems at lower levels of consumption than men, probably reflecting women's lower total body water, gender differences in alcohol metabolism, and effects of alcohol on postmenopausal estrogen levels. Mortality and breast cancer are increased in women who report drinking more than two drinks daily. Higher levels of alcohol consumption by women are associated with increased menstrual symptoms, hypertension, and stroke. Women who drink heavily also appear to have increased infertility and spontaneous abortion. Adverse fetal effects occur after variable amounts of alcohol consumption, making any alcohol use during pregnancy potentially harmful. CONCLUSIONS: In general, advising nonpregnant women who drink alcohol to have fewer than two drinks daily is strongly supported by the epidemiologic literature, although specific recommendations for a particular woman should depend on her medical history and risk factors. PMID- 9754526 TI - Nitrogen excretion and the cardiorespiratory physiology of the gulf toadfish, Opsanus beta. AB - Gulf toadfish, Opsanus beta, are facultatively ureotelic and can excrete the majority of their nitrogenous waste as urea. Urea excretion occurs in "pulses." The hypothesis that pulsatile urea excretion reflects sudden, transient, generalized increases in the branchial conductance was investigated by the simultaneous monitoring of cardiorespiratory variables, oxygen uptake, and whole body urea, ammonia, and/or 3H2O effluxes. The direct monitoring of both expired branchial water and water exiting a respirometer demonstrated that urea pulses were derived from the gills. No significant changes in ventilation or cardiac frequency, oxygen uptake, or ammonia efflux were observed during natural urea pulses, refuting the hypothesis that pulsatile urea excretion reflects pulsatile increases in the generalized diffusive properties of the gill for solute transfer. An alternative model for pulsatile urea excretion postulates that the gill urea permeability is increased periodically by the insertion and/or activation of specific urea transporters into gill cell membranes. Pulsatile urea excretion was abolished by pretreatment with the cytoskeletal-disrupting agent colchicine; colchicine may block trafficking of urea transporter-containing vesicles. Exocytosis of water following the fusion of vesicles with gill cell membranes could explain the significantly elevated 3H2O efflux observed during urea pulses. PMID- 9754527 TI - Effects of testosterone on locomotor performance and growth in field-active northern fence lizards, Sceloporus undulatus hyacinthinus. AB - The role of steroids in locomotor performance and growth was examined in free living lizards. Male northern fence lizards (Sceloporus undulatus hyacinthinus) with experimentally elevated plasma testosterone concentrations had greater sprint speed (+24%) and burst stamina (+17%) than sham-implanted males after 14 23 d in the field. This enhanced performance was associated with significant energetic costs, as the testosterone-implanted lizards had reduced growth rates, and, in a companion experiment, field-active testosterone-implanted lizards had smaller fat-body masses than controls after just 3-4 wk. These results suggest that, in addition to influencing a variety of behavioral and morphological traits, testosterone may play an important role in the regulation of locomotor performance. Also, natural levels of locomotor performance may be constrained, in part, by associated costs of elevated plasma testosterone concentrations. PMID- 9754528 TI - Trimethylamine oxide and urea synthesis in rainbow smelt and some other northern fishes. AB - Trimethylamine oxide (TMAO) and urea levels in the blood of rainbow smelt, Osmerus mordax, were previously shown to increase dramatically in winter, but the means by which these osmolytes are acquired has remained unclear. In this study, I show that the smelt can synthesize TMAO via liver trimethylamine oxidase activity and thus are not completely dependent on a dietary source of TMAO. Cold acclimatized Pacific herring, Clupea harengus, were also found to have high levels of TMAO in the blood, while individuals from a temperate-water population of herring did not. Herring also had liver TMA oxidase activity, which appeared to be due to a flavin-containing monooxygenase. In both species, TMA oxidase activity did not appear to be strongly affected by temperature. TMAO data were obtained for three other northern species (Macrozoarces americanus, Eleginus gracilis, and Platichthys stellatus), and these results, together with previously reported data, suggest that TMA oxidase activity is a necessary condition for high levels of TMAO in the blood. In the smelt, urea appears to be synthesized via uricolysis and also through the action of arginase on dietary arginine, while the ornithine urea cycle appears to be nonfunctional. There was no relation among several species of northern fishes between levels of urea in the blood and levels of uricase or arginase activity. Together, these results provide further evidence of the importance of TMAO and urea in some cold water fishes, demonstrate that the synthetic machinery for these osmolytes is present in the liver, and suggest that the elevated levels in response to cold may be due to conservation rather than to increased production. PMID- 9754529 TI - Radiative heat loss in gentoo penguin (Pygoscelis papua) adults and chicks and the importance of warm feet. AB - Adult penguins and their chicks differ considerably in their apparent body insulation. The chicks are covered in down, whereas the adults have the short, hard body feathers characteristic of the family, so mechanisms of heat loss may vary considerably between the two groups. We examined radiative heat loss by measuring body surface temperatures of gentoo penguins (Pygoscelis papua) in Antarctica. At the time the birds were considered to be in their thermoneutral zone, and there was little or no wind. Measurements of infrared emission were made on breeding adults and in large downy, and thermally independent, chicks in relation to environmental temperature. All 28 external body surface sites measured were positively correlated with ambient temperature, although there was considerable intersite variability in the relationship between site temperature and ambient temperature. Foot temperature increased most rapidly per degree ambient temperature increase, followed by the flippers, followed by the trunk. This pattern was particularly pronounced in the chicks, indicating that the exceptional heat-loss capacities of the feet may counteract for the reduced capacity of the flippers. Net heat transfer by radiation was examined using Stefan-Boltzmann's law and preliminary data on the surface area of a gentoo penguin body. This showed that between ground temperatures of 5 degrees and 15 degrees C overall heat transfer remains essentially constant, although radiative heat loss from the trunk decreases, this being counteracted by increasing heat transfer from the flippers and feet. Over the same temperature range the specific radiation heat transfer of the feet increased approximately 100 times faster per degree ambient temperature increase than did that of the flippers. This and the bimodality in foot temperature found in the study birds even under constant ambient temperatures indicate that within the thermoneutral zone heat loss by radiation in gentoo penguins is primarily executed using the feet, through which the blood circulates in pulses. PMID- 9754530 TI - Water balance, growth, development, and survival of arboreal frog eggs (Chirixalus eiffingeri, Rhacophoridae): importance of egg distribution in bamboo stumps. AB - We studied the effects of substrate moisture and flooding on the arboreal eggs of Chirixalus eiffingeri and determined the possible causes of egg mortality. Eggs appear highly permeable to water vapor, losing 16.24% and 38.38% of initial egg mass in 2 h at 90% and 45% relative humidity, respectively. Eggs that experienced positive water uptake developed faster, hatched earlier with larger hatchlings, and had greater hatching success than eggs that experienced negligible or negative water uptake. The hatching success of eggs that were submerged in water in bamboo stumps was significantly lower than that of eggs that were incubated on the water surface and was significantly correlated with the water PO2. In some bamboo stumps, we observed chironomid and tipulid larvae preying on submerged eggs. A dilution of water collected from bamboo stumps did not increase the hatching success of eggs. The water PO2 of bamboo stumps in the field was 67.4+/ 18.8 mmHg, and the degree of hypoxia of water in each bamboo stump was correlated with the turbidity. Our findings demonstrated that the vertical distribution of C. eiffingeri eggs on walls of bamboo stumps significantly influenced the growth, development, and survival of embryos. Eggs deposited too far from the water may become desiccated, while eggs deposited too close to the water may become submerged and die of hypoxia or predation by insect larvae. PMID- 9754531 TI - Sexual dimorphism in physiological performance of whiptail lizards (genus Cnemidophorus). AB - Numerous studies have examined sexual dimorphism in the morphology and behavior of vertebrates; very few, however, have explicitly investigated the possibility of gender differences in physiological performance, despite the observations of such differences in humans. In this study, I investigated physiological sexual dimorphism in the lizard genus Cnemidophorus by measuring five whole-animal traits, all of which are likely to influence fitness in these species: burst speed, endurance, maximal exertion capacity, standard metabolic rate, and evaporative water loss rate. Because at least some of these traits are known to be strongly influenced by body size, I tested for dimorphism using both absolute and size-corrected trait values. An examination of six Cnemidophorus species and subspecies revealed a strong trend toward higher absolute trait values in males for all variables except endurance. Most of the dimorphism in standard metabolic rate and evaporative water loss rate could be explained by differences in body mass between males and females; for the locomotor traits, however, body size explained only a small fraction of the overall sexual dimorphism. The portion of trait differences not explained by body size was likely due to gender differences in physiology, such as differences in relative muscularity and fat content. PMID- 9754532 TI - Osmoregulation of the Atlantic stingray (Dasyatis sabina) from the freshwater Lake Jesup of the St. Johns River, Florida. AB - The goals of this study were to (1) measure plasma osmolytes and rectal gland weights of a freshwater (FW) Atlantic stingray (Dasyatis sabina) population in the St. Johns River, Florida, and (2) determine how these parameters change after acclimation to seawater (SW). We hypothesized that the FW D. sabina may show physiological divergence from marine D. sabina, because the FW individuals reproduce and complete their life cycle in the St. Johns River. The FW D. sabina hyperregulate their plasma osmolality (621.4 mOsm kg(-1)), with plasma Na+, Cl-, and urea concentrations of 211.9, 207.8, and 195.9 mmol L(-1), respectively. FW D. sabina were exposed to 100% SW for 8 d, and their hematocrit did not change significantly compared to control animals left in FW. However, plasma osmolality increased significantly (953 mOsm kg(-1)), with significant increases in plasma concentrations of Na+, Cl-, and urea to 319.13, 296.1, and 329.76 mmol L(-1), respectively. The plasma of the SW-adapted D. sabina was hypo-osmotic and hypoionic to 100% SW. Rectal gland weight to body weight (RGBW) ratios of FW D. sabina were about 80% lower than RGBW ratios reported for marine D. sabina; the RGBW ratio did not increase significantly after SW acclimation. This may indicate that branchial and renal mechanisms are also involved with ion excretion. We conclude that the FW D. sabina are physiologically euryhaline and have not evolved the osmoregulatory strategy of stenohaline FW Potamotrygonid stingrays. PMID- 9754533 TI - Digestive responses during food restriction and realimentation in nestling house sparrows (Passer domesticus). AB - We used nestling house sparrows (Passer domesticus) under laboratory conditions to test for modulation of digestive efficiencies during periods of low and high food intake and tested the hypothesis that nestlings would exhibit compensatory changes in digestive efficiency following a period of food restriction. During the low intake period, nestlings were held at constant body mass for 48 h beginning on either day 3 or day 6 of life by feeding them at 50% of control rations. After 48 h of food restriction, nestlings were fed as much as they could consume, allowing the nestlings restricted at day 6 (early restriction not assessed) to consume 14% more food than control nestlings. For nestlings restricted at day 6 apparent dry mass assimilation of the entire diet was found to be 5% and 8% lower during food restriction and realimentation, respectively, compared with control nestlings that were not under- or overfed. There were no significant differences in radiolabeled starch assimilation efficiencies between control and restricted nestlings. Starch assimilation efficiencies remained constant from 3 d of age onward in control nestlings. Total starch extracted was lower during food restriction but reached a rate similar to that of control nestlings during the realimentation period. Passage times (time of first defecation, mean retention time, and mode passage time) measured with an indigestible marker were longer during food restriction and shorter during realimentation, relative to control nestlings. During realimentation there was no difference in intestinal rates of hydrolysis or mediated uptake of L-leucine compared with control nestlings. The main effect of changing food intake was apparently to alter flow rate, and hence retention time, causing slight changes in digestive efficiency. Thus, nestlings did not exhibit compensatory changes in digestion rates as implied by the hypothesis. Our finding of a lower dry mass assimilation efficiency and similar total starch assimilation during realimentation (relative to controls) helps explain why nestling house sparrows do not display compensatory growth, despite higher food intake. Our results indicate that the gut has little spare capacity to deal with increased food intake during growth following food restriction. PMID- 9754534 TI - Diurnal variation and effects of feeding on blood glucose in the giant tiger prawn, Penaeus monodon. AB - Previous studies on crustacea have demonstrated significant diurnal rhythms in blood glucose. However, glucose concentration in the blood of food-deprived Penaeus monodon, held in indoor or outdoor tanks, did not exhibit a diurnal rhythm under photoperiods of 8 h light and 16 h darkness (8L: 16D) or under a 13.5L: 9.5D photoperiod, with simulated or natural full moon conditions. Prawns held on photoperiods of constant light, 20L : 4D, 16L : 8D, 12L : 12D, 8L : 16D, 4L : 20D, or continuous darkness did not have significantly different mean blood glucose levels. Mean blood glucose levels varied between 0.77 and 1.39 mmol/L, depending on conditions. Pronounced and significant increases in blood glucose levels occurred within 20 min of feeding, with peak levels after 100 min. The rise in blood glucose level observed after feeding was independent of the eyestalks, and hence putative crustacean hyperglycaemic hormone, and was not from endogenous carbohydrate stores. Under appropriately controlled conditions, blood glucose concentrations can be used as an index of nutritional status in penaeid prawns. PMID- 9754535 TI - Metabolic reserves and evolved stress resistance in Drosophila melanogaster. AB - We have examined starvation and desiccation resistance in 43 outbred populations of Drosophila melanogaster that have diverged from a common ancestral population as a result of a variety of defined selection protocols. The populations differ up to 8.5-fold in desiccation resistance and up to 10-fold in starvation resistance. We used these populations to search for evolved physiological changes that might explain the differences in stress resistance. We examined two hypotheses for increased stress resistance that had been proposed previously in the literature: (1) that increments in starvation resistance are principally the result of differential lipid accumulation, and (2) that changes in glycogen accumulation play a role in evolved increases in resistance to desiccation stress. By quantifying desiccation resistance, starvation resistance, lipid content, and carbohydrate content in each of our populations of flies, we were able to demonstrate strong correlations between the capacity of the flies to resist starvation and the quantity of lipid or carbohydrate that the flies had stored. The strongest correlation (R2 = 0.99) was observed when the total energy content of both the lipid and carbohydrate stores was regressed against starvation resistance. These results demonstrate that the flies responded to selection for starvation resistance through a genetically determined increase in both lipid and carbohydrate storage. Similar analyses of the correlation between lipid storage or total energy storage and desiccation resistance revealed no significant correlations. Carbohydrate storage was significantly correlated with desiccation resistance in female but not in male flies. These results suggest that different forms of stress are resisted with distinct physiological mechanisms and that the evolutionary response of the flies to stress selection is specific to the stress imposed. PMID- 9754536 TI - Comments on a negative appraisal of taxidermic mounts as tools for studies of ecological energetics. PMID- 9754537 TI - Optic nerve dysplasia associated with meningeal defect in Sprague-Dawley rats. AB - Unilateral and bilateral dysplasias of the optic nerve (ON) were observed in 20/114 male and 14/110 female Sprague-Dawley rats at 12 weeks of age. Grossly, the intracranial segment of the affected ON had nodular thickening, bifurcation, and curvature. Nodular thickenings were seen in 20 males and 11 females. One female had a bifurcated ON. Curvature was observed in the left ONs of two females. Of 34 ON dysplasias, 12 ONs tapered off into a thin filament at the portion anterior to the dysplastic lesions. The intraorbital segments of the ONs in 33 rats were also reduced in size and were hardly recognizable in the meningeal sheath in 10 rats. Both eyeballs appeared normal in all the animals examined. Histologically, nerve fibers in intracranial and intraorbital segments of the ONs that appeared as slender filaments were markedly reduced in number. Nerve fibers in nodular thickenings were intertwined in haphazard fashion, forming scrollworklike structures. The meningeal sheaths in intracranial segments of the ONs in 15 rats and in intraorbital segments in eight rats were partially missing. The naked portion of the ON protruded into the meningeal spaces or gaps. The data indicate that developmental failures in the ON may have been induced due to insufficient blood supply through the meningeal covering or herniation of growing nerve fibers into the defective meninges. However, etiology and pathogenesis of this condition remain unclear. PMID- 9754538 TI - Cerebrospinal cuterebriasis in cats and its association with feline ischemic encephalopathy. AB - Over the past 10 years, 10 cats with primary central nervous system infection by larvae of Cuterebra flies have been documented at Cornell University. Clinical abnormalities noted in all cats were progressive and most commonly consisted of depression (6/10), blindness (6/10), and behavior changes (2/10). Affected cats presented most commonly in July (2/10) and August (7/10); one cat was presented in September. The diverse histopathologic changes are unique to this aberrant migration and consist of a combination of five characteristic features: 1) parasitic track lesion (7/10), 2) superficial laminar cerebrocortical necrosis (10/10), 3) cerebral infarction (8/10), 4) subependymal rarefaction and astrogliosis with or without ependymal cell loss (7/10), and 5) subpial astrogliosis (7/10). Changes 2-5 occurred throughout the parenchyma unassociated with the track lesion or the parasite in the affected tissue. The larvae have been recovered most commonly in the region of the olfactory bulbs and peduncles, optic nerves, and cribriform plate, suggesting entry from the nasal cavity. Many of the changes noted are suggestive of a toxic factor elaborated by the parasite and borne within the cerebrospinal fluid, as well as vascular compromise as a component in those cats with brain infarction. Because of the prevalence of infarction associated with this syndrome and the lack of reported cases of such lesions in regions of the world devoid of the fly, we propose that aberrant cuterebral larval migration in the brain is the cause of feline ischemic encephalopathy. PMID- 9754539 TI - Dependence of humoral hypercalcemia of malignancy on parathyroid hormone-related protein expression in the canine anal sac apocrine gland adenocarcinoma (CAC-8) nude mouse model. AB - Circulating parathyroid hormone-related protein (PTHrP) is the primary humoral factor in dogs with spontaneous humoral hypercalcemia of malignancy (HHM) and adenocarcinomas derived from apocrine glands of the anal sac. A canine apocrine adenocarcinoma model of HHM in nude mice (CAC-8) was developed and characterized. After 32 passages in vivo, a spontaneous variant of the tumor (CAC-8 Lo Ca) that has altered cellular morphology and that fails to induce HHM in tumor-bearing nude mice has been discovered. The hypercalcemic and nonhypercalcemic tumor lines were compared by tumor weight, effect on body weight, serum calcium concentration, plasma PTHrP concentration, histopathology, expression of PTHrP protein by radioimmunoassay and immunohistochemistry, and expression of PTHrP mRNA by in situ hybridization and northern blot analysis. Messenger RNA expression for other factors and cytokines known to alter PTHrP secretion or bone resorption in vivo, including tumor necrosis factor alpha (TNF alpha), interleukin (IL)-1, IL-6, and transforming growth factor beta (TGF beta), were also measured in the adenocarcinomas. There was no significant difference in weight of individual tumors. Nude mice bearing the CAC-8 (Lo Ca) tumor maintained normal body weight as compared with non-tumor-bearing control mice. In contrast, mice with the CAC-8 (Hi Ca) tumor had markedly decreased body weights. The CAC-8 (Hi Ca) tumor-bearing mice had severe hypercalcemia (mean = 13.4 mg/dl) and increased plasma concentrations of PTHrP (30.4 pM), whereas the CAC-8 (Lo Ca) tumor-bearing mice had a mean serum calcium concentration of 10.1 mg/dl and mildly increased PTHrP concentrations (5.7 pM) as compared with control mice (9.0 mg/dl and 1.0 pM, respectively). The original tumor (CAC-8 [Hi Ca]) is a well differentiated adenocarcinoma, whereas the variant tumor (CAC-8 [Lo Ca]) is a solid carcinoma with both polygonal and spindle-shaped cells. The CAC-8 (Lo Ca) tumor had decreased PTHrP mRNA expression and protein synthesis. Messenger RNA expression of TGF beta, TNF alpha, IL-1, and IL-6 was similar in both tumors and was consistent with the central role of PTHrP in the induction of hypercalcemia in this animal model. PMID- 9754540 TI - Macrophages, myofibroblasts, and extracellular matrix accumulation in interstitial fibrosis of chronic progressive nephropathy in aged rats. AB - Progressive renal fibrosis is considered to be the final common pathway leading to chronic renal failure. Macrophages are thought to play a role in the induction of the myofibroblasts that produce extracellular matrix (ECM) proteins in renal interstitial fibrosis. We immunohistochemically investigated the relationship between infiltrating macrophages and myofibroblast development in chronic progressive nephropathy (CPN) in 24 month-old male F344 rats, and we also analyzed components of ECM proteins using immunofluorescence microscopy. According to histomorphologic criteria for severity, described elsewhere, rats with CPN were divided into grade 1 (n = 20), grade 2 (n = 34), grade 3 (n = 10), and grade 4 (n = 6). The ratio of fibrotic tissues per unit area, determined by morphometric analysis, was increased with advancing grade of nephropathy. The number of interstitial macrophages continued to be increased gradually, with a peak in grade 4. Alpha-smooth muscle actin-positive myofibroblasts developed, surrounding the regenerating renal tubules in conjunction with the fibrotic areas. The number of the myofibroblasts was also increased, with a peak in grade 3, but in grade 4, it was slightly decreased. There was a significant relationship between the number of infiltrating macrophages and the degree of interstitial fibrosis (r = 0.802; P < 0.05). These observations suggest that macrophages and myofibroblasts might be key cells in fibrogenesis in CPN. However, there was no significant correlation between the numbers of macrophages and myofibroblasts (r = 0.198; P > 0.05), although a significant relation between these cells has been reported in the early stages of experimental rat renal fibrosis. Immunostaining for collagen type IV demonstrated increased expression in thickened tubular basement membranes. Abnormal depositions of collagen types I and III, fibronectin, and tenascin were also observed in fibrotic areas adjacent to dilated or atrophic tubules with thickened basement membranes. These ECM proteins were increased in conjunction with the grade of nephropathy, suggesting that ECM accumulation might contribute to progression of renal interstitial fibrosis. PMID- 9754541 TI - Lesions of subclinical doberman hepatitis. AB - This investigation describes histologic lesions in the livers of 18 Doberman Pinschers suffering from subclinical doberman hepatitis (DH). The dogs' ages ranged from 2.5 to 7 years; 15 were females and 3 were males. At the time of liver biopsy, the dogs had no clinical signs of liver disease, although serum alanine aminotransferase (ALT) values had been elevated in two samples in successive months. In the histologic examination, all biopsies revealed parenchymal and portal mononuclear inflammation. In the parenchyma, the inflammation was diffuse, with multifocal clusters of inflammatory cells. The periportal reaction was usually mild to moderate. Bridging necrosis (3/18) and bile duct proliferation (2/18) were rare. Excessive copper was detected by rubeinic acid stain in every specimen. Postmortem liver samples were obtained from nine dogs 3.5-65 months after the initial biopsy specimen; five of these dogs had been euthanatized for reasons other than DH, and liver specimens revealed piecemeal necrosis (5/5), bridging necrosis (3/5), and bile duct proliferation (2/5). Four of them had been euthanatized because of DH. Liver lesions of these dogs were typical for chronic active hepatitis, with bridging and piecemeal necrosis (4/4), portal expansion (4/4), bile duct proliferation (4/4), and fibrosis (4/4). A scoring system was used to evaluate changes numerically from biopsy to postmortem samples. Lesions in all dogs had progressed. The most important histologic changes were expansion of portal areas (P = 0.008), increased periportal and bridging necrosis (P = 0.008), increased fibrosis (P = 0.016), and proliferation of the bile ducts (P = 0.063). PMID- 9754542 TI - Large granular lymphocyte leukemia/lymphoma in six cats. AB - This report describes six cases of feline large granular lymphocyte lymphoma identified by light microscopy on the basis of their characteristic azurophilic granulation in Giemsa-stained plastic sections and by electron microscopy on the basis of their typical granules. Although the granules of all the tumor cells were negative for peroxidase activity, they all demonstrated chloroacetate esterase and acid phosphatase activity. All the tumors reacted with cross reacting antibodies against the CD3 antigen (epsilon chain) and did not react with a cross-reacting monoclonal antibody directed against epitopes on cytoplasmic domains of the CD20 antigen. Three tumors had a positive reaction with a monoclonal human CD57-like antibody. This is highly suggestive of either a cytotoxic T cell or a natural killer cell origin of the neoplasias. In three cats, although other abdominal organs were affected to a variable extent, the main neoplastic lesions were localized in the gastrointestinal tract and the jejunal lymph nodes. In contrast, in the other three cats, organ involvement was more widespread, affecting the lung (two), myocardium (two), precardiac mediastinum (one), salivary gland (one), and spinal cord (one); in addition, leukemia was present in two of these cats. The data presented indicate that tumors made up of large granular lymphocytes occur more frequently in cats than previously assumed and that they share many characteristic features with specific subtypes of clonal disorders of large granular lymphocytes in humans. PMID- 9754543 TI - Histomorphological and immunohistochemical studies of chronic active hepatitis in Doberman Pinschers. AB - Liver tissue samples were reviewed from 35 Doberman Pinschers with chronic active hepatitis in the precirrhotic stage. Thirty dogs had elevated hepatic copper concentrations, and five had normal liver copper concentrations. The earliest changes were inflammation and scar tissue deposition around the small hepatic vein branches. There was also apoptosis of scattered hepatocytes in zone 3. Inflammation consisted of macrophages, lymphocytes, and plasma cells. As the disease progressed, collagen deposition increased around the hepatic veins; in some liver specimens, thin scar tissue septa radiated from the hepatic vein branches, and inflammation spread to include the portal tracts. The sinusoids adjacent to the scar tissue were converted to endothelial-lined, thin-walled vessels. Chronic active hepatitis (commonly referred to as Doberman hepatitis or chronic active hepatitis of Dobermans) is a progressive fibrosis, inflammation and hepatocyte loss beginning among zone 3 hepatocytes around the terminal hepatic vein branches. The histomorphologic changes were the same among those Dobermans with elevated hepatic copper and those with normal hepatic copper. The cause was not determined, but these morphologic studies support the idea of immune-mediated disease. PMID- 9754544 TI - Comparative neurovirulence and tissue tropism of wild-type and attenuated strains of Venezuelan equine encephalitis virus administered by aerosol in C3H/HeN and BALB/c mice. AB - To assess the potential for aerosol administration of vaccines for Venezuelan equine encephalitis virus (VEE), we compared the neurovirulence and tissue tropism of the wild-type Trinidad donkey (TrD) strain to those of the attenuated TC83 and V3526 strains of VEE in mice. Six to 8-week-old female C3H/HeN and BALB/c mice were aerosol exposed to one of the three VEE strains. Three mice of each strain were euthanatized at different times and their tissues were processed and stained using hematoxylin and eosin, immunohistochemistry, and in situ hybridization. All three viral strains infected the brains of mice and induced encephalitis. TrD spread caudally from the olfactory bulbs to all regions of the brain, caused widespread necrotizing panencephalitis by day 5, and resulted in 100% mortality (geometric mean = 7 days) in both mouse strains. By comparison, TC83 relatively spared the caudal regions of the brain but still caused 100% mortality in the C3H/HeN mice (geometric mean = 12 days), yet it did not kill any BALB/c mice. V3526 infectivity of the brain was the most limited, mainly affecting the neocortex and diencephalon. This virus was not lethal in either mouse strain. The TrD strain also infected the olfactory neuroepithelium, local lymphoid tissues, teeth, and vomeronasal organs, whereas the affinity of TC83 and V3526 outside the brain was essentially limited to the olfactory neuroepithelium. Attenuated VEE strains administered to mice by aerosol have restricted tissue tropism as compared with wild-type virus; however, even attenuated strains can infect the brain and induce encephalitis. PMID- 9754545 TI - Effects of low (modified-live virus vaccine) and high (VR-2385)-virulence strains of porcine reproductive and respiratory syndrome virus on pulmonary clearance of copper particles in pigs. AB - Seventy-five 3-week-old, crossbred pigs from a herd free of porcine reproductive and respiratory syndrome virus (PRSSV) were randomly assigned to three groups: uninfected controls, pigs inoculated intranasally with RespPRRS/Repro modified live virus vaccine (RespPRRS), and pigs inoculated intranasally with a high virulence strain of PRRSV (VR-2385). Pigs were intravenously infused with 3% copper phthalocyanine tetrasulfonic acid (0.2 ml/kg) in normal saline 30 minutes before necropsy, which was performed 3, 7, 10, 14, or 28 days postinoculation (DPI) with PRRSV. There were no differences in serum copper concentration in samples collected at 0, 15, or 30 minutes after infusion. Copper concentrations in the lungs of VR-2385-inoculated pigs were significantly lower than levels in the lungs of control and RespPRRS-inoculated pigs at 7, 10, and 14 DPI (P < 0.05). The greatest difference between the groups was observed at 10 DPI. Liver and spleen copper concentrations were slightly, but not significantly, higher in both PRRSV-infected groups. The percentage of lung affected by grossly visible pneumonia ranged from 0 to 5.6% in the RespPRRS-inoculated group and from 15.2 to 46.4% in the VR-2385-inoculated group, with lesions peaking at 7 and 10 DPI, respectively. PRRSV antigen was demonstrated in both pulmonary alveolar macrophages (PAMs) and pulmonary intravascular macrophages (PIMs) by immunohistochemical methods. Copper particles were demonstrated in the PIMs by light microscopy. PRRSV was isolated from bronchoalveolar lavage fluid of VR-2385 infected pigs from 3 to 28 DPI and from RespPRRS-inoculated pigs from 7 to 28 DPI. No PRRSV, PRRSV antibodies, or PRRSV-induced pneumonia was detected in the control group. These results suggest that 1) PRRSV has a detrimental effect on the uptake of copper particles by PIMs, 2) the severity of PRRSV-induced damage to PIMs differs among strains, and 3) demonstration of PRRSV-induced decreased pulmonary clearance supports the hypothesis that PRRSV infection may make pigs more susceptible to bacterial septicemia. PMID- 9754546 TI - Pedal osteosarcoma in a donkey. AB - An 18 1/2-year-old castrated male donkey with progressively worsening right forelimb lameness presented with a mass on the distal dorsal aspect of its P3 bone. Grossly, the firm, gritty mass was infiltrative, disrupted the contours of the overlying hoof wall, and had mottled and cavitated areas on cut surface. Histologically, the growth was composed of densely cellular sheets of mildly pleomorphic mesenchymal cells forming irregularly shaped islands of poorly mineralized osteoid. The neoplastic mass had patchy areas of necrosis. The diagnostic possibilities considered for this donkey's mass include osteosarcoma, osteoma, ossifying fibroma, and fibrous dysplasia. Careful consideration of the gross and histological characteristics of this donkey's mass support a diagnosis of osteosarcoma. PMID- 9754547 TI - Aino virus antigen in brain lesions of a naturally aborted bovine fetus. AB - A bovine fetus aborted at 187 days of gestation was serologically and immunohistopathologically examined. Serum and cerebrospinal fluid samples had high titers of virus-neutralizing antibody for Aino virus. A severe necrotizing encephalopathy was noted. Aino virus antigen was demonstrated in neuroglial cells within the brain lesion. The destruction of developing neuronal cells appeared to be a significant feature of the pathogenesis of lesions due to Aino virus infection in the central nervous system. PMID- 9754548 TI - Lower motor neuron disease in the Griffon Briquet Vendeen dog. AB - Two 2-month-old Griffon Briquet Vendeen pups from the same litter were evaluated for progressive weakness and hind limb paresis. The paraparesis progressed rapidly to extensor paralysis with subsequent involvement of the forelimbs with flexor paralysis. The appendicular muscles of all four limbs became progressively atrophied. Lesions included severe loss of neurons in the ventral horns of the spinal cord, Wallerian degeneration of ventral spinal roots, and peripheral nerve and neurogenic appendicular muscular atrophy. The clinical and morphological findings were consistent with a progressive lower motor neuron disease. PMID- 9754549 TI - The phenotype of H-2M-deficient mice is dependent on the MHC class II molecules expressed. AB - For a broader view of the role of H-2M as an accessory molecule in antigen presentation, we investigated the degree to which different MHC class II isotypes and alleles depend on H-2M to function in vivo. We generated H-2M-deficient animals expressing Ek/b or Ak molecules in addition to the Ab molecules already present in the mutant strain, and compared the ability of the different MHC class II molecules to present antigen at the cell surface for recognition by T cells, and contribute to positive selection of CD4+ T cells in the thymus. Biochemical analyses were performed to assess MHC class II maturation, and to determine the peptide content of the molecules. In the absence of H-2M, Ek/b molecules contained a more heterogeneous set of class II-associated invariant chain peptides (CLIP) than Ab did, which, unlike Ab-CLIP complexes, were not SDS stable. Unlike Ab molecules, both Ek/b and Ak efficiently presented exogenously added peptides to T cells in the absence of H-2M. In addition, epitopes from some proteins, especially those known to be invariant chain independent, were presented by Ak molecules in the mutant animals. To our surprise, expression of Ek/b overcame the positive selection defect observed in H-2M-deficient mice expressing Ab alone. In contrast, Ak expression did not augment positive selection of CD4+ T cells in the mutant animals. Some of these findings in vivo contrast significantly with findings from in vitro studies on murine MHC class II molecules in human DM-deficient cell lines. PMID- 9754550 TI - Anti-IL-4 treatment prevents dermal collagen deposition in the tight-skin mouse model of scleroderma. AB - The tight-skin (Tsk/+) mutant mouse, a putative murine model of scleroderma, is characterized primarily by the excessive deposition of collagen and other extracellular matrix molecules in the dermis, and also by a developmentally acquired defect in pulmonary architecture. Passive transfer experiments have suggested an etiologic role for the immune system in Tsk/+ dermal pathology. In addition, CD4+ T lymphocytes have been shown to be required for the excessive accumulation of dermal collagen in these mice. As IL-4, a product of differentiated CD4+ T cells, is capable of regulating the synthesis of various matrix molecules (including type I collagen) by fibroblasts in vitro, we investigated the potential role of IL-4 in mediating Tsk/+ dermal fibrosis. Confirming that Tsk/+ cells are capable of responding to IL-4, we found receptors for this cytokine on Tsk/+ embryonic fibroblasts and a dermal fibroblast cell line derived from these mice. Furthermore, IL-4 receptors on Tsk/+ fibroblasts were functional since IL-4 stimulation in vitro increased type I collagen secretion from these cells. These results demonstrated the potential for IL-4 to be directly involved in the excessive deposition of dermal collagen in Tsk/+ mice. Critical insight into the role played by IL-4 in mediating the dermal phenotype, however, was obtained following the administration of neutralizing anti-lL-4 antibodies to Tsk/+ mice. This treatment prevented the development of dermal fibrosis, leading to normalization of dermal collagen content. Given the requirement for CD4+ T cells in Tsk/+ dermal fibrosis, our results suggest that Th2 cells and/or factors elaborated by this T cell subset may play a key role in regulating dermal collagen content in this strain. PMID- 9754551 TI - Efficient induction of cytotoxic CD8+ T cells against exogenous proteins: establishment and characterization of a T cell line specific for the membrane protein ActA of Listeria monocytogenes. AB - The property of listeriolysin (LLO) to introduce soluble passenger proteins into the cytosol of antigen-presenting cells allows the induction of CD8+ cytotoxic T cells against such antigens. To overcome the potential problem of presentation of the immunodominant epitope LL091-99 by H-2Kd, a variant LLO92A was established in which Tyr 92 was replaced by Ala. Immunization of BALB/c mice with purified LLO92A failed to stimulate cytotoxic T cells specific for either the epitope LLO91-99 or for any other LLO-derived peptide. Injection of mixtures of purified LLO92A and soluble nucleoprotein (NP) of influenza virus into mice resulted in a strong cytotoxic T cell response exclusively directed against NP. The LLO92A variant was successfully used to generate, propagate and characterize a CD8 T cell line specific for the membrane-bound virulence factor ActA of Listeria monocytogenes. Interestingly, wildtype ActA bound to the surface of live L. monocytogenes was not presented by MHC class I molecules to the CD8+ T cell line. PMID- 9754552 TI - Eosinophils are not required to induce airway hyperresponsiveness after nematode infection. AB - Eosinophilic inflammation of the airways is believed to play a central role in the pathogenesis of bronchial asthma. Inoculation of mice with the nematode Nippostrongylus brasiliensis induces pulmonary inflammation, characterized by a marked infiltration of eosinophils, subsequent to the migration of parasites through the lungs. Infection is associated with polarized Th2 responses in different strains of mice tested. Thus, this model may be useful to determine the relationship between established pulmonary eosinophilic inflammation, Th2 immune responses and airway changes in a nonallergic background. In the present study, we have used IL-5-deficient mice to evaluate the role of IL-5 in eosinophilic lung inflammation and airway hyperresponsiveness (AHR). In wild-type C57B/6 mice, infection with N. brasiliensis resulted in eosinophil accumulation, associated with extensive lung damage characterized by hemorrhage and alveolar wall destruction, and a strong AHR following methacholine treatment. In IL-5-deficient mice, eosinophil infiltration and the associated lung damage was abrogated. Nonetheless, AHR was unimpaired. Our results suggest that eosinophil accumulation plays a central role in lung damage but is not responsible for the induction of airway constriction following N. brasiliensis infection. PMID- 9754553 TI - Fas (CD95)-transduced signal preferentially stimulates lupus peripheral T lymphocytes. AB - Fas (CD95) is a cell surface receptor whose biological function in circulating peripheral T cells is not well understood. To address the question of abnormal T cell sensitivity to Fas stimulation in systemic lupus erythematosus (SLE), we studied Fas-transduced stimulation and apoptosis in peripheral blood T cells from patients with SLE and normal control. Immobilized anti-Fas monoclonal antibodies (mAb) (imCH-11; IgM type) significantly stimulated SLE T cell proliferation compared to T cells from normal donors and patients with rheumatoid arthritis (p < 0.003 and p < 0.005, respectively). The soluble form of CH-11 and other immobilized anti-Fas mAb (UB-2, ZB-4; IgG type) failed to stimulate lupus T cells while immobilized human Fas ligand did. Furthermore, imCH-11 induced IL-2 and IL 6 mRNA expression. However, imCH-11 activation failed to induce expression of the T cell activation surface molecules CD25 and CD69. Addition of exogenous ceramide, a second messenger for Fas-mediated apoptosis signaling, also induced T cell proliferation in SLE and normal controls. Moreover, fumonisin B1, a specific ceramide synthase inhibitor, and caspase inhibitors markedly suppressed imCH-11 induced T cell proliferation, suggesting that the ceramide pathway may be involved in Fas-transduced stimulation signals in SLE T cells. These results show that SLE T cells have an alteration in the Fas signal transduction pathway leading to cell proliferation. This defect may be important in Fas-mediated peripheral immune homeostasis. PMID- 9754554 TI - Interleukin-16, produced by synovial fibroblasts, mediates chemoattraction for CD4+ T lymphocytes in rheumatoid arthritis. AB - The massive infiltration of synovium with CD4+ T cells during the course of rheumatoid arthritis (RA) implies the expression of chemoattractant factors by resident synovial cells. Therefore, we analyzed the expression of IL-16, a potent chemoattractant for CD4+ T cells, to account for the accumulation of CD4+ T cells in RA. Indeed, IL-16 was found to be significantly elevated in synovial fluid (SF) from patients with RA as compared to non-RA arthritis (p < 0.001), osteoarthritis (p < 0.001) and controls (p < 0.001). Chemotaxis studies showed IL 16 to contribute to the strong chemotactic activities of RA-SF. In situ hybridization (ISH) revealed IL-16 mRNA-expressing cells located within the lining layer of rheumatoid synovial tissue. In the sublining area, only scattered IL-16 transcript-positive cells could be detected, mainly adjacent to blood vessels. By a double-labeling technique, combining ISH for IL-16 mRNA and immunohistochemistry for CD68, synovial fibroblast-like, CD68-negative cells were identified as a major source of IL-16 mRNA within RA synovium. This study demonstrates that synovial fibroblasts produce IL-16 in RA and thus mediate chemoattraction of CD4+ cells into synovial tissue. PMID- 9754555 TI - IL-4-regulated enteropathy in an intestinal nematode infection. AB - The relationship between intestinal pathology and immune expulsion of gastrointestinal nematodes remains controversial. Parasite expulsion is associated with intestinal pathology in several model systems and both of these phenomena are T cell dependent. Immune expulsion of gastrointestinal helminth parasites is usually associated with Th2 responses, but the effector mechanisms directly responsible for parasite loss have not been elucidated. In contrast, the intestinal pathology observed in many other disease models closely resembles that seen in helminth infections, but has been attributed to Th1 cytokines. We have used infection with the nematode Trichinella spiralis in mice defective for cytokines or their receptors to investigate cytokine regulation of both immunopathology and parasite rejection. Consistent with previous findings, we found that parasite expulsion is IL-4 dependent. Contrary to expectations, however, the enteropathy is not regulated by IFN-gamma but by IL-4. Moreover, abrogation of severe pathology in TNF receptor-defective animals does not prevent parasite expulsion. TNF is therefore involved in intestinal pathology in nematode infections, apparently under regulation by IL-4- and Th2-mediated responses. This work therefore not only reveals a novel interplay between IL-4 and TNF, but also that the IL-4-dependent protective response against the parasite operates by a mechanism other than merely the gross degradation of the parasite's environment brought about by the immune enteropathy. PMID- 9754556 TI - The role of the B cell receptor V region in peripheral B cell survival. AB - We investigate the role of the antigen-specific B cell receptor (BCR) in the establishment and maintenance of the peripheral B cell pools. We studied the fate of a population of transgenic B cells expressing a BCR without V region (Tg(deltaVmu)). We found that the Tg(deltaVmu) B cells can populate the peripheral B cell pools in the absence of other B cells, but when in the presence of a second population of non-transgenic B cells, they are virtually absent from the mature B cell compartments. By studying the rate of accumulation of 5-bromo 2'-deoxyuridine we show that the peripheral Tg(deltaVmu) B cells have a shorter life-span compared to non-transgenic B cells. By directly comparing the fate of two populations of transgenic B cells, either lacking or expressing a V region, we were able to assign the poorest competitive ability and the short peripheral survival of the Tg(delatVmu) B cells to the lack of an antigen-binding site. The results obtained support the involvement of the V region in the persistence of peripheral B cell populations. PMID- 9754557 TI - Multigenic control of lupus-associated antiphospholipid syndrome in a model of (NZW x BXSB) F1 mice. AB - In a subset of systemic lupus erythematosus (SLE) patients, antiphospholipid syndrome, characterized by occurrence of anti-cardiolipin (CL) antibodies, thrombocytopenia, thrombosis and recurrent intrauterine fetal death occurs. Male (NZW x BXSB)F1 mice, carrying the BXSB Yaa gene, serve as a model for SLE associated antiphospholipid syndrome. Using microsatellite markers in the NZW x (NZW x BXSB)F1 backcross male progeny, we mapped BXSB alleles contributing to the generation of anti-CL antibodies, platelet-binding antibodies, thrombocytopenia and myocardial infarction. Generation of each disease character was controlled by two major independently segregating dominant alleles, i.e. those on chromosomes (Chr.) 4 and 17 for anti-CL antibodies, Chr. 8 and 17 for both anti-platelet antibodies and thrombocytopenia and, to our surprise, Chr. 7 and 14 for myocardial infarction, and that a combination of the two alleles appeared to produce full expression of each character, as a complementary gene action. The alleles on Chr. 17 linked to the above three characters were all mapped in close proximity to the H-2 complex. Therefore, no single factor such as anti-CL antibodies can explain the pathogenesis of SLE-associated antiphospholipid syndrome. Rather, a combination of susceptibility alleles such as described here, along with additional modifying loci, i.e. BXSB Yaa and some from NZW, characterizes unique SLE features in male (NZW x BXSB) F1 mice. There are potentially important candidate genes which may be linked to the syndrome. PMID- 9754558 TI - Importance of a conserved TCR J alpha-encoded tyrosine for T cell recognition of an HLA B27/peptide complex. AB - Human HLA B27-restricted cytotoxic T lymphocytes (CTL) specific for the influenza A epitope NP383-391 use similar TCR alpha and beta chains, with two closely related J alpha segments used by six of nine CTL clones from three unrelated donors (Bowness et al., Eur J. Immunol. 1993. 23: 1417-1421). The role of TCR complementarity-determining region (CDR)3alpha residues 93 and 100-102 was examined by site-directed mutagenesis, following expression of the TCR alpha and beta extracellular domains from one clone as a TCR zeta fusion heterodimer in rat basophil leukemia (RBL) cells. For the first time we have measured direct binding of tetrameric HLA B*2705/NP383-391 complexes to transfected TCR. Independently peptide-pulsed antigen-presenting cells (APC) were used to induce TCR-mediated degranulation of RBL transfectants. Our results show a key role for the conserved TCRalpha CDR3 J alpha-encoded residue Y102 in recognition of HLA B27/NP383-391. Thus the Y102D mutation abolished both tetramer binding and degranulation in the presence of peptide-pulsed APC. Even the Y102F mutation, differing only by a single hydroxyl group from the native TCR, abolished detectable degranulation. Further mutations F93A and S100R also abolished recognition. Interestingly, the N101A mutation recognized HLA B27/NP in functional assays despite having significantly reduced tetramer binding, a finding consistent with "kinetic editing" models of T cell activation. Modeling of the GRb TCR CDR3alpha loop suggests that residue Y102 contacts the HLA B*2705 alpha1 helix. It is thus possible that selection of germ-line TCRAJ-encoded residues at position 102 may be MHC driven. PMID- 9754559 TI - Involvement of APO2 ligand/TRAIL in activation-induced death of Jurkat and human peripheral blood T cells. AB - The interaction of Fas with Fas ligand (FasL) mediates activation-induced cell death (AICD) of T hybridomas and of mature T lymphocytes. The TNF/TNF receptor system also plays a significant role in AICD of mature T cells and in the maintenance of peripheral tolerance. We previously demonstrated that in human Jurkat leukemia cells, AICD is triggered mainly by the rapid release of preformed FasL upon TCR stimulation. In the present work, we show that the cytotoxic cytokine APO2 ligand (APO2L; also known as TRAIL) is constitutively expressed as an intracytoplasmic protein in Jurkat T cells and derived sublines. APO2L is also detected in fresh human peripheral blood mononuclear cells (PBMC) from a significant number of donors, and the amount of both FasL and APO2L substantially increases upon blast generation. A neutralizing anti-APO2L monoclonal antibody (mAb) partially suppresses the cytotoxicity induced by supernatants of phytohemagglutinin (PHA)-prestimulated Jurkat or human PBMC on non-activated Jurkat cells, indicating that APO2L is released by these cells and contributes to AICD. A combination of neutralizing anti-APO2L and anti-Fas mAb blocks around 60 % of the toxicity associated with supernatants from PHA-activated human PBMC. These results show that FasL and APO2L account for the majority of cytotoxic activity released during AICD, and suggest that additional uncharacterized factors may also contribute to this process. PMID- 9754560 TI - Major histocompatibility complex class I presentation of exogenous soluble tumor antigen fused to the B-fragment of Shiga toxin. AB - Targeting exogenous antigen into the MHC class I-restricted presentation pathway is a prerequisite for the induction of cytotoxic T lymphocytes (CTL) which have been shown to represent an important component of the protective and therapeutic immune response to viral infections and tumors. In this study, we produced recombinant proteins composed of the receptor-binding non-toxic B-fragment of bacterial Shiga toxin derived from Shigella dysenteriae associated with an epitope from a model tumor antigen, Mage 1. We show that Shiga B-Mage 1 fusion proteins carrying an active or inactive endoplasmic reticulum retrieval signal (the C-terminal peptides KDEL or KDELGL, respectively) could be presented by peripheral blood mononuclear cells in an MHC class I-restricted manner to Mage 1 specific CTL. After pulsing B lymphoblastoid cells or dendritic cells with Shiga B-Mage 1 fusion protein, activation of the MHC class I-restricted Mage 1-specific CTL was also demonstrated. In further analysis, we showed that treatment with brefeldin A or paraformaldehyde fixation of Epstein-Barr virus-transformed B cells prevented the presentation of the Mage 1 T cell epitope, which excluded extracellular processing of the antigen. Immunofluorescence analysis also revealed that the Shiga B-Mage 1 fusion protein was largely excluded from Lamp-2 positive lysosomal structures. Therefore, the ability of Shiga toxin B-fragment to target dendritic cells and B cells and to direct antigen into the exogenous class I-restricted pathway makes it an attractive non-living and non-toxic vaccine vector. PMID- 9754561 TI - Evidence for an early appearance of modern post-switch isotypes in mammalian evolution; cloning of IgE, IgG and IgA from the marsupial Monodelphis domestica. AB - In birds, reptiles and amphibians the IgY isotype exhibits the functional characteristics of both of IgG and IgE. Hence, the gene for IgY most likely duplicated some time during early mammalian evolution and formed the ancestor of present day IgG and IgE. To address the question of when IgY duplicated and formed two functionally distinct isotypes, and to study when IgG and IgA lost their second constant domains, we have examined the Ig expression in a non placental mammal, the marsupial Monodelphis domestica (grey short-tailed opossum). Screening of an opossum spleen cDNA library revealed the presence of all three isotypes in marsupials. cDNA clones encoding the entire constant regions of opossum IgE (epsilon chain), IgG (gamma chain) and IgA (alpha chain) were isolated, and their nucleotide sequences were determined. A comparative analysis of the amino acid sequences for IgY, IgA, IgE and IgG from various animal species showed that opossum IgE, IgG and IgA on the phylogenetic tree form branches clearly separated from their eutherian counterparts. However, they still conform to the general structure found in eutherian IgE, IgG and IgA. Our findings indicate that all the major evolutionary changes in the Ig isotype repertoire, and in basic Ig structure that have occurred since the evolutionary separation of mammals from the early reptile lineages, occurred prior to the evolutionary separation of marsupials and placental mammals. PMID- 9754562 TI - Diagnosis and assessment of preclinical and clinical autoimmune encephalomyelitis using peripheral blood lymphocyte TCR. AB - In organ-specific autoimmune diseases, T cells involved in the disease development bear a particular type of TCR and infiltrate the target organ predominantly. However, it is difficult to identify disease-inducing T cells in peripheral blood lymphocytes (PBL) because such T cells are very few in number in a large pool of unrelated T cells. In the present study, we demonstrate that CDR3 spectratyping can identify experimental autoimmune encephalomyelitis (EAE) specific patterns (oligoclonal expansion of Vbeta8.2 with the shortest CDR3) in PBL at the preclinical and clinical stages of acute EAE. Analysis of nucleotide and predicted amino acid sequences of Vbeta8.2 CDR3 of spectratype-derived clones revealed that CASSDSSYEQYFGPG, which is one of the representative sequences of encephalitogenic T cell clones, constituted the predominant population in both PBL and spinal cord T cells. In chronic relapsing EAE, the EAE-specific spectratype pattern in PBL was observed during the 1 st and 2nd attacks, but not at the remission and full recovery stage. These findings indicate that the spectratyping pattern in PBL reflects the disease activity of acute and chronic relapsing EAE. Thus, CDR3 spectratyping using PBL can be used for diagnosis and assessment of T cell-mediated autoimmune diseases and is applicable to human autoimmune diseases. PMID- 9754563 TI - Rapid and coordinated switch in chemokine receptor expression during dendritic cell maturation. AB - Dendritic cells (DC) migrate into inflamed peripheral tissues where they capture antigens and, following maturation, to lymph nodes where they stimulate T cells. To gain insight into this process we compared chemokine receptor expression in immature and mature DC. Immature DC expressed CCR1, CCR2, CCR5 and CXCR1 and responded to their respective ligands, which are chemokines produced at inflammatory sites. Following stimulation with LPS or TNF-alpha maturing DC expressed high levels of CCR7 mRNA and acquired responsiveness to the CCR7 ligand EBI1 ligand chemokine (ELC), a chemokine produced in lymphoid organs. Maturation also resulted in up-regulation of CXCR4 and down-regulation of CXCR1 mRNA, while CCR1 and CCR5 mRNA were only marginally affected for up to 40 h. However, CCR1 and CCR5 were lost from the cell surface within 3 h, due to receptor down regulation mediated by chemokines produced by maturing DC. A complete down regulation of CCR1 and CCR5 mRNA was observed only after stimulation with CD40 ligand of DC induced to mature by LPS treatment. These different patterns of chemokine receptors are consistent with "inflammatory" and "primary response" phases of DC function. PMID- 9754564 TI - Acquired thymic tolerance and experimental allergic encephalomyelitis in the rat. I. Parameters and analysis of possible mechanisms. AB - Intrathymic injection of guinea pig myelin basic protein (MBP) or the immunodominant, encephalitogenic fragment of MPB, 68-86, without otherwise compromising the peripheral lymphocyte pool in adult LEW rats, dramatically inhibits onset of experimental allergic encephalomyelitis (EAE) caused by the usual peripheral inoculation with MBP in complete Freund's adjuvant. This surprising finding demonstrates that interaction of antigen and one or more components of an intact thymus can down-regulate systemic responses by mature T cells already existing in the peripheral lymphocyte pool. How this happens is not known. In studies designed to explore possible mechanisms: (a) adult thymectomized animals remain susceptible to active EAE, thus EAE cannot be attributed solely to recent thymic emigrants that might be inactivated by antigen deposited in the thymus; (b) heterotopic isografts of injected thymic lobes transfer thymic tolerance to secondary recipients, thus the tolerance effect is dominant over an intact, non-treated thymus; (c) T cells from made thymic tolerant but not immunized donors are less effective in causing EAE following adoptive transfer into, and active immunization of, secondary, irradiated recipients; and (d) animals resistant to active EAE as a consequence of thymic tolerance are fully vulnerable to adoptive EAE caused by already activated MBP specific T cell subpopulations. These results rule out a possible mechanism previously proposed for acquired thymic tolerance, i. e., that potentially pathogenic T cells traffic to the antigen-injected thymus where they are inactivated or eliminated. PMID- 9754565 TI - IL-4 is a differentiation factor for transforming growth factor-beta secreting Th3 cells and oral administration of IL-4 enhances oral tolerance in experimental allergic encephalomyelitis. AB - We have previously shown that following oral administration of myelin basic protein (MBP), regulatory T cells are generated from gut-associated lymphoid tissue and that these cells suppress experimental allergic encephalomyelitis (EAE). These regulatory T cells produce transforming growth factor-beta (TGF beta) with various amounts of IL-4 and IL-10 and these TGF-beta-secreting T cells have been termed Th3 cells. T cells in lymphoid organs drained by mucosal sites secrete IL-4 as a primary T cell growth factor. In the present study, we examined the role of IL-4 on oral tolerance and in the generation of TGF-beta secreting cells. Treatment of (PLJ x SJL)F1 mice with intraperitoneal (i. p.) IL-4 and low dose oral MBP (0.5 mg) given three times reduced the severity of EAE, whereas i.p. injection of IL-4 alone or oral MBP alone given in these suboptimal doses, showed no protection. Spleen cells from protected mice produced increased amounts of TGF-beta and reduced IFN-gamma upon stimulation with MBP in vitro. Mucosal MBP specific IgA production was significantly increased in IL-4 plus MBP fed animals. Moreover, oral administration of IL-4 (1 microg per feeding) also enhanced the suppression of EAE by oral MBP and this protective effect was reversed by administration of anti-TGF-beta antibody in vivo. Reverse transcription-PCR showed enhanced suppression of IFN-gamma in Peyer's patch in animals fed MBP and IL-4 versus those fed MBP alone. We then investigated the role of IL-4 in the generation of TGF-beta-secreting cells using MBP Ac1-11 TCR transgenic animals. Cells were cultured with IL-2, IL-4, or IFN-gamma in the presence of MBP and limiting dilution analysis for cytokine-secreting cells performed. We found that IL-4, but not IL-2 or IFN-gamma, generated TGF-beta-secreting T cells from naive splenic T cells and that these cells provided help for IgA production. These findings demonstrate that IL-4 is a differentiation factor for TGF-beta-secreting Th3 cells and oral IL-4 has a synergistic effect on low-dose oral tolerance that is associated with increased TGF-beta secretion. PMID- 9754566 TI - Split tolerance to a viral antigen expressed in thymic epithelium and keratinocytes. AB - When expressed as a transgene from the keratin 14 (K14) promoter in an MHC class II-deficient mouse, I-Ab expressed in thymic cortical epithelium promotes positive but not negative selection of I-Ab-restricted CD4+ T cells (Laufer, T. M. et al., Nature 1996. 383:81-85). Transgenic mice expressing the E7 protein of human papilloma virus 16 from the K14 promoter were studied to determine the consequence of expression of a cytoplasmic/ nuclear protein from the K14 promoter. K14E7-transgenic mice express E7 in the thymus and skin without evidence for autoimmunity to E7. Repeated immunization of FVB(H-2q) or F1(C57BL/6JxFVB) mice with E7 elicited similar antibody responses to the defined B cell epitopes of E7 in K14E7-transgenic and non-transgenic animals. In contrast, for each genetic background, a single immunization with E7 elicited demonstrable T cell proliferative responses to the major promiscuous T helper epitope of E7 in the transgenic but not the non-transgenic animals. Further, E7 immunized non-transgenic F1 (FVBxC57BL/6J) animals developed strong E7-specific cytotoxic T lymphocyte (CTL) responses and were protected against challenge with E7+ tumors, whereas similarly immunized K14E7-transgenic animals had a markedly reduced CTL response to E7 and no E7-specific tumor protection was observed, although the antibody and CTL response to ovalbumin was normal. Expression of E7 protein as a transgene from the K14 promoter in the skin and thymus thus induces E7-specific tolerance in the cytotoxic T effector repertoire, together with expansion of the E7-specific T helper repertoire. These findings demonstrate that limited tissue distribution of an autoantigen may result in "split" tolerance to that autoantigen. PMID- 9754567 TI - Altered peptide ligands and wild-type peptide induce indistinguishable responses of a human Th0 clone. AB - The response of the human influenza hemagglutinin (HA)-specific T helper clone HA1.7 to its wild-type (wt) HA307-319 peptide ligand and related altered peptide ligands (APL) was examined over a wide range of antigen concentrations. The time course of cytokine production and surface expression of CD40 ligand and CD3 was followed at the single-cell level by flow cytometry and compared with the induction of proliferation. We observed that the APL induced responses that were indistinguishable from those induced by the wt HA ligand, albeit at higher antigen densities. Moreover, the activation parameters were induced with identical kinetics. Blocking of CD4 co-ligation inhibited the recognition of the weakly stimulatory APL, but not of the wt HA307-319 peptide. Finally, in all cases the response of the T cell clone correlated with down-regulation of surface TCR/CD3 complexes. Together, these observations support a quantitative model of T cell activation. PMID- 9754568 TI - Negative regulation of Ig gene rearrangement by a 150-bp transcriptional silencer. AB - We previously showed that the V-J intervening sequence of the chicken lambda immunoglobulin locus contains a strong silencer that acts both on transcription and rearrangement. We show here that the transcriptional silencer activity can be ascribed to a minimal 150-bp fragment. The rearrangement silencing activity was previously shown by the replacement of the V-J intervening sequence with a neutral DNA fragment that dramatically increased the rate of rearrangement of the transgene. Insertion of the minimal silencer in this neutral fragment is shown here to result in a marked decrease in rearrangement of the transgenic construct. Strikingly, deletion of 28 bp from the 150-bp fragment abolished most of the transcriptional silencing activity and had a similar effect on rearrangement. These results conclusively correlate the silencing activity on both rearrangement and transcription. PMID- 9754569 TI - Induction and functional characterization of beta2-microglobulin (beta2-mu)-free HLA class I heavy chains expressed by beta2-mu-deficient human FO-1 melanoma cells. AB - The frequent loss of beta2-microglobulin (beta2-mu) in malignant cells has stimulated interest in the functional characteristics of beta2-mu-free HLA class I heavy chains, since this information contributes to assess the impact of beta2 mu abnormalities on the interaction of malignant cells with immune cells. Therefore, the present study has investigated the ability of beta2-mu-free HLA class I heavy chains to modulate NK cell-mediated lysis of melanoma cells and to present melanoma-associated antigen (MAA)-derived peptides to HLA class I restricted, MAA-specific cytotoxic T lymphocytes (CTL). Beta2-mu-free HLA class I heavy chains were induced on beta2m null FO-1 cells by sequential incubation with IFN-alpha for 48 h at 37 degrees C and for 24 h at 26 degrees C. Transfection of cells with a wild-type H-2Ld gene (FO-1Ld) enhanced the induction of beta2-mu free HLA class I heavy chains under such experimental conditions. Beta2-mu-free HLA class I heavy chains expressed on the cell membrane did not protect the B2m null FO-1 cells from NK cell-mediated lysis. Furthermore, FO-1 cells which express beta2-mu-free HLA-A2 heavy chains following transfection with a wild-type HLA-A2 gene were not lysed by HLA-A2-restricted, MAA-specific CTL lines and clones. These results indicate that association with beta2-mu is required for interaction of HLA class I molecules with NK inhibitory receptors and for peptide presentation to CTL. PMID- 9754570 TI - In the absence of IL-12, the induction of Th1-mediated protective immunity by the attenuated schistosome vaccine is impaired, revealing an alternative pathway with Th2-type characteristics. AB - Vaccination of mice with irradiated Schistosoma mansoni larvae confers high levels of immunity which is mediated by Th1-type lymphocytes. To investigate a possible role for IL-12 in the induction of protection, we have compared the immune response of IL-12 p40-deficient (KO) mice and their C57BL/6 (WT) counterparts following vaccination. Cultured lymph node cells from KO mice had markedly altered cytokine profiles with significantly decreased production of IFN gamma increased IL-4. Correspondingly, KO mice had enhanced levels of IgE. After challenge, cells recovered from the lungs of KO mice secreted abundant IL-4 and IL-5 but little IFN-gamma, while flow cytometric and histological analysis of lung cell populations recorded a very high proportion of eosinophils. The levels of protection in KO mice were substantially lower than in their WT counterparts, demonstrating the importance of IL-12 and Th1-mediated immune responses. This conclusion is reinforced by the administration of rIL-12 to KO mice immediately after vaccination which led to increased IFN-gamma and the restoration of protective immunity. Nevertheless, the data also indicated that the limited levels of protection induced in KO mice occur via an IL-12-independent pathway, possibly mediated by Th2 cells. PMID- 9754571 TI - hBRAG, a novel B cell lineage cDNA encoding a type II transmembrane glycoprotein potentially involved in the regulation of recombination activating gene 1 (RAG1). AB - The different display reverse transcription-PCR (DD RT-PCR) technique was used to identify novel cDNA detecting mRNA transcripts co-expressed with human recombination activating gene-1 (RAG1). A 5.0-kb transcript detected by the differential display amplicon 3G1 was found to correlate strongly with RAG1 mRNA expression in various human cell lines. Subsequent screenings of a pre-B cDNA library with 3G1 led to the identification of a complete cDNA we have termed hBRAG (human B-cell RAG-Associated Gene). The hBRAG cDNA encodes a 503-amino acid (aa) protein with no known homology to any nucleotide or protein sequence. The predicted molecular mass of 55 kDa was confirmed by in vitro translation. Based on sequence analysis, the predicted open reading frame encodes for a type II transmembrane spanning glycoprotein with the N-terminal 81 -aa in the cytoplasm, a 17-aa transmembrane domain, and a C-terminal 405-aa extracellular domain with four potential N-glycosylation sites. Northern blot analysis indicated a close association of the 5.0-kb hBRAG mRNA transcript with RAG1 in numerous human pro B, pre-B and mature B cell lines assessed, but not in human T cell lines. In human tissues, hBRAG is expressed at highest levels in B cell-enriched tissues, but is not expressed in fetal or adult thymus. Southern blotting analysis revealed that this gene is conserved across eukaryotes, is expressed as a single copy in the human genome, and is likely not a multigene family member. The hBRAG gene was localized to the long arm of chromosome 10 (10q26). Transfection of the full-length hBRAG cDNA increased levels of human RAG1 transcripts in the B cell line OCI LY8-C3P, but not in the non-lymphoid line K562, suggesting a B cell specific role for the hBRAG product in regulating RAG expression. PMID- 9754572 TI - HLA-E-bound peptides influence recognition by inhibitory and triggering CD94/NKG2 receptors: preferential response to an HLA-G-derived nonamer. AB - The HLA-E class Ib molecule constitutes a major ligand for the lectin-like CD94/NKG2 natural killer (NK) cell receptors. Specific HLA class I leader sequence-derived nonapeptides bind to endogenous HLA-E molecules in the HLA defective cell line 721.221, inducing HLA-E surface expression, and promote CD94/NKG2A-mediated recognition. We compared the ability of NK clones which expressed either inhibitory or activating CD94/NKG2 receptors to recognize HLA-E molecules on the surface of 721.221 cells loaded with a panel of synthetic nonamers derived from the leader sequences of most HLA class I molecules. Our results support the notion that the primary structure of the HLA-E-bound peptides influences CD94/ NKG2-mediated recognition, beyond their ability to stabilize surface HLA-E. Further, CD94/ NKG2A+ NK clones appeared more sensitive to the interaction with most HLA-E-peptide complexes than did effector cells expressing the activating CD94/NKG2C receptor. However, a significant exception to this pattern was HLA-E loaded with the HLA-G-derived nonamer. This complex triggered cytotoxicity very efficiently over a wide range of peptide concentrations, suggesting that the HLA-E/G-nonamer complex interacts with the CD94/NKG2 triggering receptor with a significantly higher affinity. These results raise the possibility that CD94/NKG2-mediated recognition of HLA-E expressed on extravillous cytotrophoblasts plays an important role in maternal-fetal cellular interactions. PMID- 9754573 TI - Lymphocyte triggering via L-selectin leads to enhanced galectin-3-mediated binding to dendritic cells. AB - For proper immune surveillance, naive lymphocytes are recruited from the blood into secondary lymphoid organs. L-selectin expressed on lymphocytes plays an important role in the initial attachment of these cells to high endothelial venules (HEV) in lymph nodes. Previously, we found that triggering via L-selectin resulted in activation of lymphocytes, followed by an alteration in their adhesion capacity. This suggested that L-selectin triggering might play a role in cell-cell interactions after lymph node entry. Here, we identify a novel adhesion mechanism involving L-selectin-triggered lymphocytes and dendritic cells, and we show that enhanced binding to dendritic cells is mediated by galectin-3 and not by integrins. Furthermore, it was shown that L-selectin-triggered T lymphocytes exhibited enhanced proliferation in an allogeneic mixed lymphocyte reaction. It is concluded that, in addition to a role for L-selectin in tethering and rolling on endothelium, triggering of the molecule on the lymphocyte surface leads to changes that are pertinent for the function of the cell after passing the HEV. We argue that the described adhesion mechanism plays a role in optimizing the initial interaction between dendritic cells and lymphocytes. PMID- 9754574 TI - Impaired MHC class I (H-2Dd)-mediated protection against Ly-49A+ NK cells after amino acid substitutions in the antigen binding cleft. AB - The MHC class I molecule H-2Dd (Dd) acts as a ligand for the inhibitory NK cell receptor Ly-49A. We have constructed altered Dd molecules by site-directed mutagenesis, replacing residues with the corresponding amino acids from the Db molecule, which fails to inhibit via Ly-49A. Mutations at positions 73 and 156 (DdS73WD156Y) impaired the protective effect of the Dd molecule, as evaluated by testing lymphoma cells transfected with the mutant gene for sensitivity to killing by Ly-49A+ NK cells in vitro and rejection by NK cells in vivo. The altered residues form a hydrophobic ridge across the floor of the antigen binding cleft. A mutation in the alpha helix of the alpha2 domain, facing the solvent and without direct contact with the peptide (DdA150S) had no effect. Dd recognition by Ly-49A+ NK cells is considered to be peptide dependent, but not peptide specific. Our results indicate that alterations of residues buried in the antigen binding cleft can induce changes in peptide binding patterns and/or conformational changes in the Dd molecule that make the trimolecular complex less permissive for inhibition of Ly-49A+ NK cells. PMID- 9754575 TI - Thymic heterotypic cellular complexes in gene-targeted mice with defined blocks in T cell development and adhesion molecule expression. AB - Thymocytes form unique multicellular complexes with epithelial cells (thymic nurse cells, TNC) and rosettes (ROS) with macrophages, epithelial cells and dendritic cells. To investigate the role of differentiation checkpoints in the formation of the thymic heterotypic complexes in vivo, we used mutant mice which have genetically defined blocks at early and late stages of T cell development. We show that RAG-1-/-, TCRbeta-/- , and p56lck-/- mice lack thymocyte ROS formation with epithelial cells, macrophages, or dendritic cells. TNC formation was not affected by TCRbeta and p56lck gene mutations but partially decreased in RAG-1-/- mice, indicating that TNC are the earliest thymocyte-stromal cell complexes formed in development, whereas ROS only appear after thymocytes have rearranged and expressed a functional TCRbeta chain. Genetic blocks in CD8 lineage commitment (CD8-/- and IFN regulatory factor-1-/- mice) and positive and negative T cell selection (CD45-/-, TCRalpha-/-, and CD30-/- mice) did not affect thymocyte-stromal cell complexes. Surprisingly, CD4-/- mice, but not MHC class II /- mice, had significantly reduced numbers of TNC and ROS, in particular, a severe defect in ROS formation with thymic dendritic cells. The CD4-/- block in ROS and TNC formation was rescued by the introduction of a human CD4 transgene. Moreover, we show that the adhesion receptors CD44 and LFA-1 cooperate in the formation of the thymic microenvironment. These results provide genetic evidence on the role of defined stages in T cell development and adhesion molecules on thymocyte/stromal cell interactions in vitro. PMID- 9754576 TI - TNF modulates susceptibility to UVB-induced systemic immunomodulation in mice by effects on dermal mast cell prevalence. AB - The mechanisms by which UV radiation is immunosuppressive are controversial, but there is growing evidence that processes of UVB-induced suppression of the immune response towards a sensitizing antigen are different if this antigen is applied to irradiated compared with non-irradiated sites. Consistent with this is our recent observation (Hart et al., J. Exp. Med. 1998. 187: 2045-2053) that the prevalence of dermal mast cells determines the extent of susceptibility of different mouse strains to UVB-induced systemic, but not local, immunosuppression. Using C57BL/6 and BALB/c mice exposed to low and high doses of UVB, respectively, in the presence of a polyclonal anti-TNF antibody, we found that TNF is directly involved as a mediator in the suppression by UVB of local immune responses. To determine whether TNF indirectly regulates UVB-induced systemic immunomodulation by altering the prevalence of dermal mast cells, dermal mast cell numbers in gene-targeted mice deficient in TNF or TNF receptors (p55/p75-/- mice) were quantified by video image analysis. A reduced dermal mast cell prevalence in these mice correlated with decreased susceptibility for systemic immunosuppression caused by UVB. We hypothesize that TNF is one molecule that controls dermal mast cell prevalence by as yet unknown mechanisms. However, it is the mediators released from mast cells upon UVB-induced degranulation, which do not include TNF, that directly signal suppressive events relevant to systemic immunosuppression. PMID- 9754577 TI - Anergic T cells actively suppress T cell responses via the antigen-presenting cell. AB - We here show that anergic T cells are active mediators of T cell suppression. In co-culture experiments, we found that anergic T cells, derived from established rat T cell clones and rendered anergic via T cell presentation of the specific antigen (Ag), were active inhibitors of T cell responses. Anergic T cells inhibited not only the responses of T cells with the same Ag specificity as the anergic T cells, but were also capable of efficiently inhibiting polyclonal T cell responses directed to other epitopes. This suppression required close cell cell contact between antigen-presenting cells (APC), anergic T cells and responder T cells, and only occurred when the epitope recognized by the anergic T cell was present. The suppression was not caused by passive competition for ligands on the APC surface, IL-2 consumption, or cytolysis, and was not mediated by soluble factors derived from anergic T cells that were stimulated with their specific Ag. When responder T cells were added 24 h after co-culturing anergic cells in the presence of Ag and APC, T cell responses were still suppressed, indicating that the suppressive effect was persistently present. However, anergic T cells were not able to suppress responder T cells that had already received a full activation signal. We propose that suppression by anergic T cells is mediated via the APC, either through modulation of the T cell-activating capacity of the APC (APC/T cell interaction), or by inhibition of T cells recognizing their ligand in close proximity on the same APC (T/T cell interaction). PMID- 9754578 TI - Innate resistance to infection by intracellular bacterial pathogens differs in mice selected for maximal or minimal acute inflammatory response. AB - The intensity of nonspecific immune reaction and the host resistance to facultative intracellular pathogens are found to be associated in lines of mice selected for maximal (AIRmax) or minimal (AIRmin) acute inflammatory reactivity. AIRmax are more resistant than AIRmin mice to Salmonella typhimurium and Listeria monocytogenes infection, the differences between lines in LD50 being > 1000 and 100 times, respectively. This difference was shown to be related to the initial bacterial containment at the infectious focus, and to the control of bacterial multiplication in the spleen during the 1st week after s. c. inoculation of the bacteria. Specific immune responses were not deeply affected by the selective process: antibody production and delayed-type hypersensitivity were both of similar intensity in AIRmax and AIRmin mice. The differential susceptibility to infection seems independent of the Nramp-1 locus polymorphism; therefore, these two lines represent a powerful model for investigating the role of other genetic loci regulating the nonspecific immunity effectors in the course of infectious diseases. PMID- 9754580 TI - IFN-gamma induces the high-affinity Fc receptor I for IgG (CD64) on human glomerular mesangial cells. AB - The deposition of immune complexes, followed by activation of complement and/or Fc receptors and generation of chemoattractants, is the most common feature of human glomerulonephritis. Recently we have shown that primary cultured human glomerular mesangial cells (HMC), which are normally negative for IgG Fc receptors, can be stimulated to express the low-affinity FcgammaRIII-A receptor isoform. In this study we further demonstrate that activation of HMC through IFN gamma resulted in the functional expression of the high-affinity Fc receptor for IgG (FcgammaRI, CD64). IFN-gamma-dependent induction of classical FcgammaRIa1 mRNA as well as a2, b2 splice variants were evident after 24 h in proliferating HMC and after 48 h in resting HMC. Transcription of FcgammaRI mRNA was also induced by IL-10 in proliferating HMC, whereas other cytokines such as IL-3, transforming growth factor-beta1 and granulocyte-macrophage colony-stimulating factor were not effective. Cell surface expression of FcgammaRI could be detected by flow cytometric analysis after IFN-gamma stimulation and was accompanied by the augmentation of MHC class II and the up-regulation of intercellular adhesion molecule-1 expression. Triggering of HMC by cross-linking FcgammaRI with F(ab')2 fragments of the anti-CD64 monoclonal antibody 22 led to enhanced synthesis of mRNA for the chemokines IL-8 and monocyte chemoattractant protein-1, indicating that the FcgammaRI of HMC is functionally active. These in vitro data suggest that engagement of both FcgammaRI and FcgammaRIII-A on activated HMC through IgG immune complexes may result in an increased chemoattraction of leukocytes into the glomerulus, contributing to the development of glomerulonephritis. PMID- 9754579 TI - A mimotope defined by phage display inhibits IgE binding to the plant panallergen profilin. AB - Birch pollen and mugwort pollen allergies are often associated with hypersensitivity to plant foods. This clinical and serological cross-reactivity is mediated by IgE antibodies reacting with homologous proteins in pollen and food. Cross-reacting homologs of the important birch pollen allergen Bet v 2 (profilin) could be detected in other pollen, fruits, nuts, and vegetables, such as celery tuber. We purified IgG/IgE antibodies from the serum of an exclusively profilin-allergic patient using affinity columns either coupled with protein extracts from mugwort pollen, birch pollen, or celery tuber. Constrained and unconstrained random nonapeptide libraries were pooled and screened with the anti profilin antibody preparations to define cross-reactive ligands. Specific ligands were enriched by successive panning rounds using the profilin-specific antibodies in series. After the last panning round enriched phage clones were screened with purified profilin-specific antibodies and IgE-binding clones were sequenced. Five out of eight positive clones (62.5 %) displayed the same circular peptide CAISGGYPVC. This peptide was synthesized and examined for its ability to inhibit IgE binding to blotted mugwort pollen, birch pollen, or celery tuber profilin. Inhibition studies showed reduction of IgE binding to profilins in all three protein extracts. As the sequence of the mimotope did not show any homology to the known birch profilin sequence this peptide is considered to mimic a common conformational IgE epitope for these examined profilins. PMID- 9754581 TI - Low expression of CD40 and B7 on macrophages infiltrating UV-exposed human skin; role in IL-2Ralpha-T cell activation. AB - After UV exposure of skin, epidermal Langerhans cells (LC) are depleted, whereas CD11b+CD36 CD1a- monocytes/macrophages (UV-Mphi) infiltrate. Different immunological outcomes in vivo are mediated by LC (sensitization) and UV-Mphi (tolerance) which may be related to the distinct T cell activation states that these antigen-presenting cells (APC) induce. We previously demonstrated that CD4+ T lymphocytes activated by UV-Mphi are, in contrast to LC-activated T cells, IL 2Ralpha deficient, and we hypothesize that this differential T cell activation is related to differences in co-stimulatory molecules between UV-Mphi and LC. Using four-color flow cytometry, we found a reduced capacity to up-regulate expression of the important co-stimulatory molecules CD40, B7-1 and B7-2 by UV-Mphi relative to LC. This alteration in co-stimulatory molecule expression was selective, because UV-Mphi express equal levels of ICAM-1 and ICAM-3, and increased levels of LFA-1, relative to LC. After bidirectional signaling with T cells during alloantigen presentation, UV-Mphi still exhibited less CD40 and B7-1 than LC. Addition of IFN-gamma induced CD40 and B7-1 expression on UV-Mphi and restored IL 2Ralpha expression on UV-Mphi-activated T cells but had no effect on IL-2Ralpha on resting or LC-activated T cells. The restoration of IL-2Ralpha expression on UV-Mphi-activated T cells by IFN-gamma was inhibited (67 %, p = 0.005) by addition of neutralizing anti-CD40. Therefore, differences in co-stimulatory molecule expression, in particular CD40, on UV-Mphi and LC are critical in determining the distinct T cell activation induced by these APC. PMID- 9754582 TI - Development of the early B cell population in Xenopus. AB - Like mammals, the amphibian Xenopus uses combinatorial joining of the immunoglobulin V, D and J elements and multiple rearrangements to generate its B cell repertoire. Xenopus larvae hatch 2 days after fertilization and individuals are under pressure to develop an immune repertoire when the number of available cells is small (approximately 5 and 200 IgM-positive cells on days 5 and 11 after fertilization, respectively). In the liver, in a first phase of differentiation spanning days 5-12 after fertilization before immunological competence, the heavy (H) chain locus starts rearranging followed by the light (L) chain locus 3 days later. By immunohistology the first B cells expressing H and L chain are detectable on day 10. Despite the small number of cells available and the lack of external antigen selection at these early stages, the repertoire is heterogeneous. The VH families are used stepwise, although their genes are interspersed in the genome. The earliest family used (VH1) is homologous to the VH3 family of human and to the VH7183 of the mouse which are also overrepresented in early mammalian development. In the second phase, from day 12-13 onwards, the spleen differentiates and the animal becomes immunologically competent. The V, D and J usage is similar to that of adults although VDJ junctions lack N nucleotides until metamorphosis. A preferential reading frame for D and one specific DJ junction are overrepresented during this second phase. The visible bias toward homology-based junction results in fact from selection after rearrangement. PMID- 9754583 TI - Hyporesponsiveness to lipopolysaccharide alters the composition of NF-kappaB binding to the regulatory regions of inducible nitric oxide synthase gene. AB - Repeated exposure to bacterial endotoxin causes a diminished response by the host to further exposure. One important feature of this hyporesponsiveness is a reduced macrophage production of nitric oxide (NO) via the inducible nitric oxide synthase (iNOS) pathway. Using a murine macrophage model, we observed that hyporesponsiveness was accompanied by a decrease in the levels of NO release (measured as nitrite), iNOS protein and iNOS gene transcription. The expression of the putative lipopolysaccharide (LPS) receptor, CD14, was not altered. In vivo genomic footprinting showed that the same binding sites are occupied in the iNOS promoter and enhancer of desensitized macrophages and of LPS-responsive macrophages, yet the composition of NF-kappaB in the nuclei of these cells was found to be altered. The transcriptionally inactive homodimer p50-p50 represented the predominant binding activity in nuclei from LPS-pretreated cells before and after stimulation. Nuclei from cells which had not been pretreated but were stimulated contained more of the transcriptionally active p50-p65 heterodimer than their pretreated counterparts. Consistent with this, the cytosolic steady state level of an inhibitor of NF-kappaB activity, I-kappaBalpha, was decreased in normal cells but not in pretreated cells. We propose that the presence of an overwhelming excess of transcriptionally inactive p50 homodimers on their kappaB sites in the iNOS control region in pretreated cells may block kappaB site binding by p50-p65, thereby reducing the activity of the protein complex governing iNOS transcription. PMID- 9754585 TI - Reversible left ventricular dysfunction in the setting of coronary occlusive disease. PMID- 9754584 TI - Human IgA- and IgM-secreting intestinal plasma cells carry heavily mutated VH region genes. AB - Single IgA- or IgM-secreting plasma cells were isolated from histological sections of human jejunum and terminal ileum, and Ig heavy chain variable (VH) region genes were amplified and sequenced. Taken together, 62 of 63 cells analyzed harbored somatically mutated VH region genes, indicating that the vast majority of both IgA- and IgM-secreting intestinal plasma cells derive from germinal center B cells. On average, rearranged VH genes of IgA- and IgM secreting plasma cells showed a mutation frequency of 9.0 % and 8.5 %, respectively, which exceeds the level of somatic mutation of V region genes carried by human memory B cells. Moreover, we detected deletions or insertions in the complementarity-determining regions of 5 of the 58 functional VH region genes analyzed, suggesting that these alterations may contribute to the diversification of the human antibody repertoire in the course of an immune reaction. PMID- 9754586 TI - Factors influencing exercise performance in patients with left ventricular dysfunction. SOLVD Investigators. Studies of Left Ventricular Dysfunction. AB - BACKGROUND: The determinants of exercise performance are multifactorial and incompletely understood in patients with symptomatic left ventricular (LV) dysfunction, with much less information regarding asymptomatic LV dysfunction. This study assessed the hemodynamics and neurohormonal factors influencing exercise performance in patients with LV ejection fractions > or =0.35, both symptomatic and asymptomatic, enrolled in Studies of LV Dysfunction. METHODS AND RESULTS: We studied 103 patients enrolled prospectively in Studies of LV Dysfunction before randomized therapy; 38 were symptomatic and 65 had no or minimal symptoms. By using rest-exercise gated equilibrium radionuclide ventriculography and cuff blood pressure, we assessed the heart rate, LV and right ventricular (RV) volumes and ejection fractions, total peripheral resistance, the LV peak systolic pressure/end systolic volume ratio as an index of contractility, and plasma renin and norepinephrine at rest and during maximal graded supine bicycle ergometer exercise. Changes between rest and exercise were evaluated as indices of cardiovascular reserve. The cumulative workload ranged from 120 to 2,100 watt-min. At rest, the LV ejection fraction was 0.30 in asymptomatic patients and 0.25 in symptomatic patients, respectively (P < .0004). During exercise, asymptomatic patients had greater increases in heart rate, systolic blood pressure, LV ejection fraction, and cardiac output than symptomatic patients (P > or = .05). Combining all patients, the strongest univariate correlates of exercise workload were the ability to increase heart rate (r = 0.70), the pressure/volume ratio (r = 0.63), and systolic blood pressure (r = 0.55), and to decrease the total peripheral resistance (r = -0.47) with moderate correlations for the ability to increase LV and RV ejection fractions (r = 0.33 and 0.35, respectively) (P < .0008). By multivariate analysis, workload was modeled best by the changes in four factors: heart rate, systolic blood pressure, and the LV and RV ejection fractions (R2 = 0.54, P < .001). CONCLUSION: Exercise performance and its hemodynamics differed in patients with symptomatic and asymptomatic LV dysfunction. Rather than features at rest, the reserve capacities for increasing heart rate, systolic blood pressure, and the LV and RV ejection fractions were the predominant cardiac mechanisms related to greater exercise performance. PMID- 9754587 TI - Predicting short-term outcome in severely ill heart failure patients: implications regarding listing for urgent cardiac transplantation and patient selection for temporary ventricular assist device support. AB - BACKGROUND: The purpose of this study was to determine which patients on a cardiac transplantation list required a ventricular assist device. METHODS AND RESULTS: In a preliminary study, 26 patients with decompensated severe New York Heart Association class IV chronic heart failure were studied. Blood levels for sodium, hemoglobin, cytokines, neurohormones, and hemodynamics were obtained. During short-term follow-up of 40 days, 12 patients had undergone emergent implantation of a ventricular assist device (range 1-27 days, mean 5 days), 4 died (range 14-38 days, mean 26 days), and 5 were alive and receiving only medical therapy while waiting for a transplantation. In addition, five patients had undergone transplantation (range 5-29 days, mean 18 days, excluded from further analysis). Survival curves were constructed by comparing the incidence of death and the implantation of an emergent ventricular assist device in patients with values of a variable above or below the mean value (or median for nonnormally distributed data). There was a significantly greater incidence of death or need for a ventricular assist device in patients with higher levels of tumor necrosis factor-alpha (P = .008), lower levels of serum sodium and hemoglobin (P = .02 and P = .03, respectively), higher heart rates (P = .03), and higher plasma norepinephrine levels (P = .01). The Cox proportional hazards model demonstrated that only serum sodium (P = .03) independently predicted those patients who died or who required emergent left ventricular assist device. CONCLUSION: Numerous variables, particularly serum sodium, need to be considered when evaluating which patients on the transplant list require early assist device implantation or urgent transplantation. These preliminary observations merit confirmation in a larger patient population. PMID- 9754588 TI - Early reduction in plasma norepinephrine during beta-blocking therapy with metoprolol in chronic heart failure. AB - BACKGROUND: The possible role exerted by modulation of sympathetic outflow in the clinical effects of beta-blockade in chronic heart failure was tested during short- and long-term treatment. METHODS AND RESULTS: Oral metoprolol (30-150 mg/day) was added to conventional therapy in 14 patients with idiopathic dilated cardiomyopathy, left ventricular ejection fraction (LVEF) of <0.45, and New York Heart Association class II or III. Norepinephrine plasma levels, which are an index of sympathetic activation, decreased by 27.57 +/- 18.03% after 1 month (P < .005), but returned to pretreatment levels after 6 months. LVEF increased by 7.7 +/- 6.0 ejection fraction units after 6 months (P < .005 vs baseline and P < .05 vs 1 month). Long-term beta-blockade resulted in nonsignificant improvements in functional class, symptom score, and oxygen consumption at peak exercise. After 1 month, the reduction in plasma norepinephrine levels and the changes in LVEF were inversely correlated (P < .01). No other correlation emerged during short- or long-term treatment. CONCLUSION: In conclusion, the reduction in plasma norepinephrine levels during short-term beta-blockade was not proportional to the clinical benefits and may have been attributed to the direct inhibition of sympathetic outflow. The early reduction in circulating norepinephrine levels may decrease cardiac performance through withdrawal of sympathetic support when the favorable effects of beta-blockade have not had time to occur. The role that sympathetic modulation may exert in the long-term clinical benefits of metoprolol deserves further investigation. PMID- 9754589 TI - Angiotensin-converting enzyme gene expression in skeletal muscle in patients with chronic heart failure. AB - BACKGROUND: Skeletal muscle factors may influence functional limitation in patients with heart failure. The renin-angiotensin system is activated in chronic heart failure. Treatment with angiotensin-converting enzyme (ACE) inhibitors improve symptoms and prognosis. The goal of this study was to quantify and localize skeletal muscle ACE-mRNA in patients with chronic heart failure and in control subjects, and to elucidate skeletal muscle fiber area and capillary density. METHODS AND RESULTS: Biopsies from the lateral vastus muscle were taken from 9 patients before and after treatment with enalapril and in 10 control subjects. ACE-mRNA was quantified with reverse transcription polymerase chain reaction. Immunohistochemistry was used to localize ACE within skeletal muscle. No difference in ACE-mRNA transcripts between patients and control subjects was detected, nor did ACE gene expression change after treatment with enalapril. The number of ACE-mRNA transcripts was related to muscle fiber area, whereas an inverse relationship between the number of ACE transcripts and capillary density was found. ACE was detected in the endothelial cells of capillaries in skeletal muscle. CONCLUSION: ACE is expressed in skeletal muscle and is confined to endothelial cells. The close relationship between capillary density and number of ACE transcripts indicate that activation of the renin-angiotensin system has an impact on capillary growth. PMID- 9754590 TI - Effects of short-term forearm exercise training on resistance vessel endothelial function in normal subjects and patients with heart failure. AB - BACKGROUND: Exercise training improves endothelium-dependent vasodilation in animals. This study was designed to determine whether forearm exercise training improves endothelium-dependent vasodilation in control subjects and patients with heart failure, a disease associated with abnormal endothelium-dependent vasodilation. METHODS AND RESULTS: We used strain gauge plethysmography to assess the effects of short-term forearm exercise training on resistance vessel function in 11 control subjects and 7 patients with New York Heart Association class II and III heart failure. Subjects performed 30 minutes of handgrip exercise four times a week for 4-6 weeks. In the control subjects, exercise training increased forearm blood flow (FBF) responses to intra-arterial acetylcholine (20 microg/min) from 6.9 +/- 3.1 to 12.2 +/- 3.0 mL/min/100 mL and peak reactive hyperemic FBF responses from 38.1 +/- 5.6 to 47.4 +/- 5.6 (P < .05). Basal FBF and responses to nitroprusside, L-N-monomethyl arginine and acute forearm exercise were not significantly changed. In the patients with heart failure, chronic forearm exercise did not significantly change any of the above-measured parameters. CONCLUSION: Forearm exercise training improves endothelium-dependent vasodilation and peak hyperemic FBF in control subjects but not in patients with heart failure. These data suggest that resistance vessel abnormalities may not be as readily modifiable by exercise training in patients with heart failure compared with control subjects. PMID- 9754591 TI - Functional impact of an increase in ventricular mass after myocardial damage and its attenuation by converting enzyme inhibition. AB - BACKGROUND: A increase in left ventricular mass after ventricular damage has been identified as an initial response to injury. However, the functional significance of this response has not been clearly established and is the focus of this study. METHODS AND RESULTS: Twelve mongrel dogs underwent transmyocardial direct current shock to produce transmural left ventricular damage. Six were assigned to converting enzyme inhibitor therapy initiated 24 hours after damage and continued for 4 weeks. The remaining six dogs served as a control group. Left ventricular structure (mass and end diastolic volume) and systolic function (regional and global ejection fraction at rest and during afterload stress) were assessed by magnetic resonance imaging before damage and at the end of the 4-week period. After myocardial damage, left ventricular mass increased from 93.6 +/- 4.0 to 107.5 +/- 3.4 gm in the control group (P < .01) with no change in ventricular volume. Ramipril-treated dogs displayed a reduction in mass (83.2 +/- 2.2 to 74.6 +/- 2.9 gm, P < .05). In the control group, there was greater reduction in global ejection fraction in response to afterload stress at 4 weeks compared with baseline (-16 +/- 4 vs -4 +/- 3%, P = .03). Ejection fraction response to afterload stress was maintained at 4 weeks in the converting enzyme inhibitor treated group (-5 +/- 3 vs - 1 +/- 4%) and was different at 4 weeks from the control group (-1 +/- 4 vs -16 +/- 4%, P = .004). CONCLUSION: The increase in left ventricular mass noted after direct current shock was associated with the impairment of systolic function during afterload stress. Inhibition of this mass increase results in preservation of function, thus further supporting the concept that attenuation of ventricular remodeling should be a therapeutic goal. PMID- 9754592 TI - Angiotensin II blockade followed by growth hormone as adjunctive therapy after experimental myocardial infarction. AB - BACKGROUND: Recombinant human growth hormone (rhGH) has shown beneficial effects on cardiac function after myocardial infarction (MI) in rats. High-dose angiotensin II (AT1) receptor blockade in normal rats inhibited the hypertrophic effect of growth hormone (GH), therefore we investigated whether GH effects after MI would be enhanced by giving it in sequence after remodeling had been inhibited by prior AT1 blockade (losartan, L). METHODS AND RESULTS: Rats given losartan for 10 weeks after MI followed by rhGH for 2 weeks (2 mg/kg twice a day, GH plus losartan) were compared with rats given losartan for 10 weeks followed by placebo for 2 weeks (placebo plus losartan group) and with untreated controls (n = 17 20/group). Average MI sizes and left ventricular (LV) end diastolic (ED) dimensions (echocardiography) did not differ between groups. In GH and losartan, body weight (BW) was increased but left ventricular weight (LVW)/BW was reduced, and the LV fractional shortening and LV dP/dtmax (catheter tip micromanometer) were increased compared with the control group (20.3 vs 15.4% and 5579 vs 4699 mmHg/s, respectively, P < .05). The cardiac index also was significantly increased. In the placebo plus losartan group, the LVW/BW was also reduced and the cardiac index increased versus controls. Stroke volume was increased in GH plus losartan group compared with both placebo plus losartan and controls, and the systemic vascular resistance was significantly decreased only in the GH plus losartan group. The ED posterior wall thickness (noninfarcted wall) was increased in GH plus losartan compared with both control and placebo plus losartan. Left ventricular end diastolic pressure reduction was not significant in GH plus losartan group versus controls but was reduced in placebo plus losartan group, whereas LV relaxation (tau) was improved in both groups versus control rats. Thus, persistent remodeling effects caused by prior AT1 blockade undoubtedly contributed to some responses, but short-term GH given in sequence after chronic AT1 blockade had favorable actions on the failing heart and peripheral circulation by increasing LV wall thickness with partial reversal of unfavorable remodeling, lowering of vascular resistance, improvement of LV contractility, and enhanced LV systolic function and cardiac index relatively late after experimental MI. PMID- 9754593 TI - Consecutive screening and enrollment in clinical trials: the way to representative patient samples? AB - BACKGROUND: Trials usually do little or nothing to ensure random samples of patients representative of the disease under investigation. METHODS AND RESULTS: To evaluate the effects of consecutive screening and enrollment, we compared the demographic characteristics of patients from three interventional trials, applying consecutive screening with those from nine similar survival trials of risk-stratified patients with either myocardial infarct or congestive heart failure. Administrative information was also compared. Patients in the consecutive studies were 5-10 years older and more often female. Six of the nine conventional studies enrolling high-risk patients had a 1-year mortality of < or =10% compared with 22-28% in the consecutive studies. The conventional studies that accounted for screenings tended to screen four to six times more patients, but typically enrolled fewer of those screened: 4-7% versus 17-26% in the consecutive studies. The conventional studies excluded the majority of those eligible, up to 85% versus 35% in consecutive studies. The conventional studies enrolled less than 1 patient per center per month compared with two to four times as many in the consecutive studies. CONCLUSIONS: Patients in conventional cardiovascular trials, not using consecutive screening are selected and not representative of the disease under investigation. They are younger, more often male, and have lower risk. This makes such trials less reliable and useful. It prolongs the length of a trial study and makes it more expensive. Consecutive screening and enrollment are feasible and offer a detailed description of the patient selection The consecutive principles contribute to representative patient samples and should be mandatory in future clinical trials. PMID- 9754594 TI - Consecutive screening and enrollment in clinical trials. PMID- 9754595 TI - Strategies for patient recruitment into clinical trials. PMID- 9754596 TI - To screen or not to screen: how to improve clinical trials. PMID- 9754597 TI - Validity versus generalizability in clinical trial design and conduct. PMID- 9754598 TI - Clinical trials in heart failure: can we expect the results to be replicated in clinical practice? PMID- 9754599 TI - Role of nitric oxide in ventricular dysfunction. PMID- 9754600 TI - Acute renal failure: controversies, clinical trials, and future directions. AB - Acute renal failure (ARF) is an important clinical syndrome. Despite the frequent occurrence of ARF, nephrologists have not made major therapeutic inroads in the treatment or prevention of ARF. This article will speculate as to why it has been so difficult to gain a substantial foothold in the ongoing battle against ARF. First, some of the major controversies regarding the pathogenesis of ARF will be considered. Scientific debates regarding the mechanisms of ARF have greatly enriched the scientific literature, but may have slowed the development of clinically applicable therapies. Controversies regarding the treatment of ARF will then be discussed. Next, the fate of several recent clinical trials in ARF will be examined. Finally, the future directions that research in ARF may pursue will be contemplated. PMID- 9754601 TI - Mechanisms of cellular injury in ischemic acute renal failure. AB - Significant advances have been made in understanding the pathophysiology of injury at the cellular level in ischemic acute renal failure. Alterations in the actin cytoskeleton are of central importance to the structural, physiological, and biochemical changes that occur in proximal tubule cells during acute ischemic injury. These cytoskeletal alterations occur rapidly and are dependent on the severity and duration of ischemic injury. Most importantly, alterations in the actin cytoskeleton are responsible for changes in the cell surface membrane that modify cell polarity, cell-cell interactions, and cell-matrix interactions. Ultimately, these cytoskeletal alterations play a major role in the decrement in glomerular filtration rate that is the hallmark of ischemic acute renal failure. PMID- 9754602 TI - The role of neutrophils in acute renal failure. AB - The role of neutrophils in acute renal failure is controversial. Acute renal failure can clearly occur in the absence of neutrophils. However, recent studies using specific neutrophil markers indicate that neutrophils accumulate in postischemic kidneys. Moreover, reperfusion of ischemic kidneys with neutrophils worsens ischemic injury and causes kidney neutrophil retention. Neutrophil retention is dependent on the state of neutrophil activation and the duration of renal ischemia. This interaction could account for the high frequency of acute renal failure in conditions associated with prolonged prerenal asotemia and neutrophil priming such as the adult respiratory distress syndrome, or sepsis. Neutrophil retention is mediated by interaction of neutrophil integrins and endothelial cell ICAM-1 because maneuvers reducing the expression and/or function of these adhesion molecules is protective in experimental models of ischemia. Nitric oxide is a key modulator of neutrophil worsening of ischemic injury because maneuvers that decrease nitric oxide production worsen and those which increase nitric oxide protect ischemic kidneys from neutrophil effects. The clinical significance of neutrophils may relate to the observation that bioincompatible membranes activate complement, and retard recovery from acute renal failure. In conclusion, neutrophils are an important contributor to ischemic acute renal failure. It remains to be determined whether decreasing neutrophil function accelerates recovery in acute renal failure. PMID- 9754603 TI - Necrosis and apoptosis in acute renal failure. AB - Renal tubular cells that are lethally injured after an acute ischemic or nephrotoxic insult to the kidney can die by necrosis or apoptosis. Necrosis is usually the result of overwhelming and severe cellular ATP depletion. In contrast, there are many potential causes of apoptosis in acute renal failure (ARF). These include cytotoxic events not severe enough to induce necrosis, a relative deficiency of renal growth factors, and loss of cell-matrix or cell-cell adhesive interactions. In some situations, receptor-mediated events induced by tumor necrosis factor-alpha (TNF-alpha) or Fas (CD95) may play a role in apoptosis in ARF. Necrosis and apoptosis are distinct morphologically and biochemically. Necrosis results in an early loss of plasma membrane integrity, the release of injurious substances from the cytosol, and an inflammatory reaction in the surrounding tissue that is readily detected morphologically. In contrast, apoptosis is characterized by progressive cell shrinkage with condensation and fragmentation of nuclear chromatin. Apoptotic cells ultimately break up into plasma membrane-bound vesicles called "apoptotic bodies" that are rapidly phagocytosed by macrophages and neighboring epithelial cells. In experimental models of ARF in vivo, apoptosis of renal tubular cells has been shown to occur in two distinct phases. The first phase of apoptosis occurs early on, between 12 and 48 hours after the acute ischemic or nephrotoxic insult. The second phase of apoptosis occurs many days later, during the recovery phase of ARF. Tubular cell apoptosis occurring shortly after the acute insult probably contributes to tubular cell loss and the tubular dysfunction associated with ARF. In contrast, the apoptosis associated with the recovery phase has been postulated to contribute to the remodeling of injured tubules and to facilitate their return to a normal structural and functional state. Therapeutic interventions that inhibit or promote apoptosis of renal tubular cells have the potential for minimizing renal dysfunction and accelerating recovery after ARF. PMID- 9754604 TI - Mechanisms of renal repair and survival following acute injury. AB - The reaction of the renal epithelium to injury is heterogenous. Some cells die, others survive apparently intact, while others commit to repair. The determinants of these responses appear to depend on signal transduction pathways and molecular responses that is segment specific and interactive. The kidney, as do cells in culture exposed to various noxious stimuli, react in a typical manner referred to as the stress response. The response is comprised of kinases and their molecular targets as well as cell cycle-specific factors that determine whether a cell survives the injury or not. We propose that this response can be modified by survival factors which upregulate those aspects of the response that are cytoprotective and which downregulate those that are cytoreductive. Preliminary data will be presented to demonstrate the feasibility of this approach. PMID- 9754605 TI - Acute renal failure: prevention and nondialytic therapy. AB - Acute renal failure (ARF) is a common illness with significant associated mortality and morbidity. Despite the advent of renal replacement therapy and the advancement in dialytic technology, the mortality of ARF has not significantly changed in the last 30 years. The cost of treating acute renal failure with the available therapies inflicts a tremendous financial burden on the health care system. The majority of patients with acute renal failure have multiple etiologies which are frequently iatrogenic. Physicians frequently underestimate the level of renal dysfunction in patients and therefore interventions to curb or treat renal failure are delayed. It is clear that ARF can be averted with more vigilance and early interventions. No pharmacological agent has yet been approved for the treatment of acute renal failure. Several substances are in the various stages of animal and human trials. Until one becomes available for use in the treatment of renal failure, it is clear that prevention is the principal element in the management of ARF. The purpose of this review is to discuss the various risk factors for acute renal failure, methods of prevention, and pharmacological interventions that may be beneficial in the treatment of ARF. PMID- 9754606 TI - Dialysis and related therapies. AB - Dialysis continues to be the major supportive intervention in severe acute renal failure (ARF). The impact of dialysis on outcomes from ARF is best viewed from an historical perspective when comparisons can be made with the predialysis era. The overall effect of dialysis therapy on survival has been limited primarily to reducing death from direct renal-related causes such as hyperkalemia and pulmonary edema. Attempts to improve survival in ARF patients requiring dialysis has been focused in areas in intensity of therapy, dialysis modalities, and ultrafiltration membranes. Optimal choices in these areas continue to be controversial with additional controlled studies required. Because of the continuing high mortality in ARF regardless of specific dialysis practices, a number of recent studies have examined the value of outcome predictors in choosing patients for dialysis. This is another complex arena in which there are still insufficient data to predict in all settings the exact probabilities of survival. Pediatric dialysis for ARF infrequently receives discussion in primarily adult-oriented critical care. There has been an increasing refinement in dialysis indications, access, and modalities in recent years including the use of continuous dialysis in infants and children. PMID- 9754607 TI - Outcomes in acute renal failure. AB - Although patients with acute renal failure (ARF) are now older and sicker than in the past, mortality remains constant or even slightly lower, which suggests a better management of the syndrome. Several clinical conditions, mainly assisted respiration, hypotension, oliguria, coma and jaundice, have a detrimental effect on outcome. Previous health status, original disease, a hospital and/or ICU start of the ARF, and age of the patient also seem to affect outcome of these patients. ARF observed in the ICU setting has a poorer prognosis than the ARF treated in other hospital areas. This is because of the higher number of associated organ failures observed in the ICU. Estimation of outcome could be done either using specific ARF or general ICU score systems. They allow risk stratification of the patients, and some of them give an individual prognosis that at present should not be used for a withdrawal decision. Functional outcome of ARF is usually good, although some patients need to be maintained on chronic dialysis. PMID- 9754608 TI - Radiocontrast-induced nephropathy. AB - Radiocontrast-induced nephropathy (RCIN) is a common cause of acute renal failure. It results from an ischemic injury to the medullary portion of the kidney secondary to intense renal vasoconstriction. Patients with pre-existing renal insufficiency, diabetes, congestive heart failure, taking certain medications, and exposed to large volumes of high osmolar contrast are at increased risk of developing RCIN. Prophylaxis of high-risk patients by careful hydration and avoidance of exacerbating factors can minimize the incidence of RCIN. PMID- 9754609 TI - Acute renal failure complicating muscle crush injury. AB - Extensive skeletal muscle injury, whether caused by mechanical crush or by extreme physical exertion, is incompatible with life, unless treated early and vigorously. The immediate cause of morbidity is leakiness of the sarcolemmal membrane to cardiotoxic or nephrotoxic cations and metabolites (K, PO4, myoglobin and urate) of the sarcoplasma, and rapid massive uptake by the muscles of extracellular fluid, sodium and calcium, leading to profound hypovolemic and hyocalcemic shock. Casualties who survive the early steep of hyperkalemia and arterial hypotension are susceptible to myoglubinuric acute renal failure owing mainly to the combination of renal vasoconstriction, nephrotoxicity, and tubular obstruction by myoglobin plugs and urate. Management includes immediate (prehospital) intravenous volume replacement followed by mannitol-alkaline diuresis. The alkali regimen ameliorates the acidosis associated with shock and the hyperkalemia, and protects against the nephrotoxicity of myoglobin and urate by alkalinization of the urine. Mannitol, through its impermeant hyperoncotic properties, decompresses and mobilizes muscle edema and promotes renal tubular flow, thus flushing myoglobin plugs and enhancing urinary elimination of nephrotoxic metabolites. With this regimen and when necessary also with the use of dialysis, a substantial salvage of lives, limbs, and kidney function has been achieved recently compared with invariable mortality for casualties who were buried for 3 to 4 hours or more in the early 1940s (World War 2). PMID- 9754610 TI - Functional residual capacity measurement during tracheal gas insufflation. AB - OBJECTIVE: Tracheal gas insufflation (TGI) is considered an adjunctive method to enhance carbon dioxide elimination during permissive hypercapnia in patients with acute respiratory distress syndrome. Due to increasing tidal volume and/or expiratory resistance, TGI may cause intrinsic PEEP (PEEPi), and may lessen the advantages of permissive hypercapnia. There is no reliable method to measure PEEPi during TGI. Using an argon washout method to evaluate dynamic hyperinflation, we developed a method to measure FRC with TGI flow. METHODS: We measured FRC during TGI by washing out both the ventilator and TGI circuit with 100% oxygen (O2) previously equilibrated with 10% argon and 90% O2. To test the accuracy of our system, we measured the volume in a model lung composed of two flasks. The FRC of the model lung was changed by varying its volume of water, to active 500, 1000, and 1500 mL. The change of FRC (deltaFRC) of the model lung was measured at a flow of 0, 4, 8, and 12 L/min. Then the FRC of a bellows-type model lung was measured at the same TGI flow. PEEPi of the model lung was also recorded as the pressure inside the bellows at end-expiration. RESULTS: Our FRC measurements were accurate within 10% except for that of 500 mL without TGI (12.7%+/-1.1%). As inspiratory time (TI) and/or TGI flow increased, the FRC of the bellows-type model lung increased. PEEPi and deltaFRC showed a positive correlation (r = 0.843, p < 0.001). The higher the TGI flow, the greater was the deltaFRC with both continuous and expiratory-phase TGI. FRC during continuous TGI was higher than during expiratory-phase TGI especially during long TI and high TGI flow. CONCLUSIONS: The system developed in this study can be used as a method to detect air-trapping during TGI. PMID- 9754611 TI - Use of a neonatal blood pressure cuff to monitor blood pressure in the adult finger--comparison with a standard adult arm cuff. AB - BACKGROUND: There are few suitable methods for monitoring blood pressure continuously (or intermittently) for research in adult stroke patients, who are ill but do not justify invasive intensive care monitoring. METHOD: We tested a neonatal arm blood pressure in adults by placing it on the forefinger ("finger cuff"). We compared the repeatability of the finger cuff with blood pressure measured by a standard adult arm cuff using the oscillometric technique in 168 ambulatory outpatients attending a cerebrovascular disease clinic. RESULTS: The mean difference between sequential mean blood pressure readings with the finger cuff was 0.55 mm Hg (95% confidence interval (CI) -14.36 to 15.47 mm Hg), and for the arm cuff was 3.31 mm Hg (95% CI -23.33 to 16.71 mm Hg). Measurements made with the arm cuff were shown to affect subsequent arm cuff readings made within a few minutes of the first. The mean difference between the finger cuff and arm cuff mean blood pressure readings was 0.03 mm Hg (95% CI -26.07 to 26.14 mm Hg) and agreement was better when the blood pressure was measured with the finger cuff first rather than the arm cuff. However, although there was no difference in the mean blood pressure recordings both systolic and diastolic blood pressure measurements differed systematically between arm and finger cuff. CONCLUSION: The reproducibility of sequential blood pressure measurements made with the finger cuff was better than with the arm cuff. The performance of the finger cuff compared with that of the arm cuff was sufficiently good to encourage use of the finger cuff in research involving automatic intermittent monitoring to observe sequential blood pressures over time in stroke patients. However, measurements of systolic and diastolic pressure were not the same with the two cuffs and further work on calibration of the finger cuff would be useful. PMID- 9754612 TI - Nocturnal body movements and hypoxemia in middle-aged females after lower abdominal surgery under general anesthesia: a study with the static-charge sensitive bed (SCSB). AB - OBJECTIVE: The aim of this study was to evaluate the feasibility of the static charge-sensitive-bed (SCSB) combined with pulse oximetry (SpO2) for postoperative monitoring and to determine variables which could be used for evaluating the quality of postoperative sleep and breathing. METHODS: The frequency of body movements and the perioperative breathing abnormalities were assessed using the SCSB and pulse oximeter in 15 female ASA-class I-II patients undergoing elective lower abdominal surgery under general anesthesia. Anesthesia and control of postoperative pain followed standard practice. The patients were monitored during one preoperative and three consecutive postoperative nights. Movements were analyzed according to their duration and time interval. The effect of opioids was evaluated by measuring arterial oxyhemoglobin saturation (SpO2) with pulse oximetry for one hour before and two hours after administration of standard doses of oxycodone. RESULTS: The total movement time per hour increased during the first postoperative night (p = 0.003). Conversely, periodic movement activity decreased significantly during the three postoperative nights (p = 0.05, p < 0.001, p = 0.007). The mean SpO2 decreased during the first postoperative night (95.5% vs. 94.2%, p = 0.002), but returned to the preoperative level during the following nights. No episodes of apnea with significant oxygen desaturation (a decrease in SpO2 > 5%) were observed. Opioid administration was associated with decreased mean SpO2 (94.8% vs. 93.6%, p = 0.02), but did not lead to clinically significant hypoxemia (lowest observed SpO2 89.8%). CONCLUSIONS: Postoperative periodic movement activity was suppressed, but sleep remained fragmented with frequent body movements. In our middle-aged non-obese females (ASA I-II), no severe postoperative hypoxemia was observed during the three-nights postoperative survey. Perioperative movement monitoring with the SCSB was a valuable tool in rejecting movement artefacts of SpO2 and in evaluating general sleep quality. PMID- 9754614 TI - Applying human factors to the design of medical equipment: patient-controlled analgesia. AB - OBJECTIVE: Medical instruments commonly have poorly designed user interfaces that promote human errors with life-threatening consequences. The primary hypothesis of this study was that a specific user interface could be made safer and more efficient if redesigned using human factors techniques and principles. METHODS: The user interface of a commercially available patient-controlled analgesia (PCA) pump, the Abbott Lifecare 4100 PCA Plus II infuser, was evaluated using a cognitive task analysis of bench tests and field observations. Based on this analysis, the user interface was redesigned. Important elements of the new design include a dialog structure with fewer steps, a dialog overview showing the user's location in the programming sequence, better command feedback, easier error recovery, and clearer labels and messages. The changes were evaluated by comparing a computer prototype of the new interface with a computer simulation of the old one. Twelve student nurses performed six programming tasks with each interface. Task completion time, number of errors, and subjective mental workload were collected for each trial. RESULTS: The results showed significantly faster programming times (F(1,11) = 6.85, P < 0.025), lower mental workload ratings (chi2(1) = 4.45, p < 0.025, one-tailed), and fewer errors (chi2(1) = 3.33, p < 0.05, one-tailed) with the new interface. CONCLUSION: Adopting a human factors approach to redesigning the PCA interface led to significantly faster, easier, and more reliable performance. These findings have important implications for improving the design of other computer-based medical equipment. PMID- 9754613 TI - Adaptive lung ventilation (ALV) during anesthesia for pulmonary surgery: automatic response to transitions to and from one-lung ventilation. AB - Adaptive lung ventilation is a novel closed-loop-controlled ventilation system. Based upon instantaneous breath-to-breath analyses, the ALV controller adjusts ventilation patterns automatically to momentary respiratory mechanics. Its goal is to provide a preset alveolar ventilation (V'A) and, at the same time, minimize the work of breathing. Aims of our study were (1) to investigate changes in respiratory mechanics during transition to and from one-lung ventilation (OLV), (2) to describe the automated adaptation of the ventilatory pattern. METHODS: With institutional approval and informed consent, 9 patients (33-72 y, 66-88 kg) underwent ALV during total intravenous anesthesia for pulmonary surgery. The ALV controller uses a pressure controlled ventilation mode. V'A is preset by the anesthesiologist. Flow, pressure, and CO2 are continuously measured at the DLT connector. The signals were read into a IBM compatible PC and processed using a linear one-compartment model of the lung to calculate breath-by-breath resistance (R), compliance (C), respiratory time constant (TC), serial dead space (VdS) and V'A. Based upon the results, the controller optimizes respiratory rate (RR) and tidal volume (VT) such as to achieve the preset V'A with the minimum work of breathing. In addition to V'A, only PEEP and FIO2 settings are at the anesthesiologist's discretion. All patients were ventilated using FIO2 = 1,0 and PEEP = 3 cm H2O. Parameters of respiratory mechanics, ventilation, and ABG were recorded during three 5-min periods: 10 min prior to OLV (1), 20 min after onset of OLV (II), and after chest closure (III). Data analyses used nonparametric comparisons of paired samples (Wilcoxon, Friedman) with Bonferroni's correction. Significance was assumed at p < 0.05. Values are given as medians (range). RESULTS: 20 min after onset of OLV (II), resistance had approximately doubled compared with (1), compliance had decreased from 54 (36-81) to 50 (25-70) ml/cm H2O. TC remained stable at 1.4 (0.8-2.4) vs. 1.2 (0.9)-1.6) s. Institution of OLV was followed by a reproducible response of the ALV controller. The sudden changes in respiratory mechanics caused a transient reduction in VT by 42 (8-59)%, with RR unaffected. In order to reestablish the preset V'A, the controller increased inspiratory pressure in a stepwise fashion from 18 (14-23) to 27 (19-39) cm H2O, thereby increasing VT close to baseline (7.5 (6.6-9.0) ml/kg BW vs. 7.9 (5.4 11.7) ml/kg BW). The controller was, thus, effective in maintaining V'A. The minimum PaO2 during phase II was 101 mmHg. After chest closure, respiratory mechanics had returned to baseline. CONCLUSIONS: Respiratory mechanics during transition to and from OLV are characterized by marked changes in R and C into opposite directions, leaving TC unaffected. The ALV controller manages these transitions successfully, and maintains V'A reliably without intervention by the anesthesiologist. VT during OLV was found to be consistently lower than recommended in the literature. PMID- 9754615 TI - Can capnography detect bronchial flap-valve expiratory obstruction? AB - OBJECTIVE: We have previously shown in a mechanical lung model [1] that bronchial flap-valve expiratory obstruction results in sequential lung expiration, best detected by prolonged and low magnitude tracheal expired flow (V) from the obstructed lung. However, the normal expiratory resistance of clinical ventilation circuits might also generate prolonged, low value exhaled V, that could be confused with bronchial flap-valve obstruction. We reasoned that bronchial flap-valve obstruction would also cause sequential CO2 unloading from each lung and result in a biphasic tracheal capnogram. METHODS: To test this hypothesis, we ventilated (VT, 650 ml; f, 10 br/min) a dual mechanical test lung, with each side connected to a separate alcohol-burning chamber. An airway adapter monitor system measured airway V, P, PCO2, and FO2. The circumference of the diaphragm in a respiratory one-way valve was trimmed to generate unidirectional resistance to expiratory V. Measurement sequences were repeated after this flap valve was interposed in the left "main-stem bronchus." RESULTS AND DISCUSSION: During moderate or severe left bronchial flap-valve obstruction, left bronchial V was delayed so that the left lung anatomical dead space (devoid of CO2) mixed with normal right exhalate to depress the expiratory upstroke or early plateau of the tracheal capnogram. During severe obstruction, decreased perfusion of the left lung caused lower alveolar PCO2. Then, prolonged low V from the left bronchus also resulted in depression of the end of the tracheal alveolar plateau. In general, the low magnitude of bronchial V from the obstructed lung limited its effect on the tracheal capnogram and the best marker of sequential lung emptying during bronchial flap-valve obstruction may be late exhaled V without reduction in total tidal volume. PMID- 9754616 TI - Performance of a plastic optical fiber stylet for tracheal intubation of a dog. AB - OBJECTIVE: We set out to establish whether a novel plastic optical fiber incorporated into an endotracheal tube (ETT) stylet could be used for intubation of a dog. A secondary objective examined the need for a direct illumination source from a laryngoscope. Lastly, the fragility of the system was tested. METHODS: An anesthetized dog was repeatedly intubated using a laryngoscope to elevate the tongue and the view of the larynx conducted through the plastic optical fiber stylet (placed within an endotracheal tube) and displayed on a television monitor. Four prototype identical stylets were tested. Repeated intubations were attempted with each stylet and graded as either successful or failed. All four stylets were tested 10 times each using a Miller 4 blade and direct illumination from the laryngoscope. Two of the four stylets were reused during an additional 10 intubation attempts using a Miller 4 blade and laryngoscope (without batteries) with only ambient light. Finally, one stylet was used for intubation after 10, 20, 30, 40 and 50 sharp 90 degree bend-and straighten cycles using a Miller 4 blade and laryngoscope for direct illumination. RESULTS: All attempted intubations were successful. However, the image quality was dramatically better when direct illumination from a laryngoscope was used than when ambient light was used. One plastic optical fiber stylet was successfully used to intubate after having been used for 20 intubations and 50 sharp 90 degree bend-and-straighten cycles. A partial lens separation occurred between the 41st and 50th bend cycle but the image remained adequate enough to successfully intubate again. CONCLUSIONS: A novel plastic optical fiber incorporated into an ETT stylet can be used with a laryngoscope for intubation of a dog. Direct illumination from a laryngoscope provides a better television monitor image than when only ambient light is used. The system was durable, withstanding over 20 uses and 40 sharp bend-and-straighten cycles before a lens separation failure occurred. PMID- 9754617 TI - Transcutaneous renal function monitor: precision during unsteady hemodynamics. AB - OBJECTIVE: Hospital acquired renal dysfunction, most commonly caused by renal hypoperfusion, dramatically increases mortality in intensive care patients. Glomerular filtration rate (GFR) is rapidly altered during renal hypoperfusion, and a more rapid means of GFR measurement may prompt institution of renal specific therapy. We hypothesized that a transcutaneous renal function monitor can rapidly and accurately assess acute changes in GFR within a time frame much shorter than the 2-4 hours currently available. METHODS: The study design was a prospective determination of the capability to measure GFR transcutaneously. In three different studies, concurrent transcutaneous measurement of GFR, using the rate of disappearance of 99mTc-diethylenetriaminepentaacetic acid (DTPA), was compared by correlation and standard deviation (SD) to reference standards of DTPA plasma clearance, serum inulin clearance, or serum creatinine. RESULTS: Continuous transcutaneous clearance (TC) measurement correlated with standard DTPA plasma clearance techniques (r = 0.93). Acute pharmacologically induced changes in GFR are detectable by TC measurement within 12-20 min, a time interval significantly affected by the data acquisition interval. Excess patient movement in the ICU patients created clearance artifacts in 50% of clearance traces. Retrospective analysis of ICU patient data reveal TC measurements are 93% specific and 92% sensitive for serum creatinine levels in critically ill patients. CONCLUSIONS: TC monitoring provides prompt indication of directional changes in GFR and may provide the clinician warning of inadequate resuscitation. Prospective analysis of the specificity, sensitivity, and TC guided renal specific resuscitation is needed. PMID- 9754618 TI - A cross-validated multifactorial index of perioperative risks in adults undergoing anaesthesia for non-cardiac surgery. Analysis of perioperative events in 26907 anaesthetic procedures. AB - OBJECTIVE: To develop a severity index of anaesthetic risk that predicts relevant perioperative adverse events in adults. DESIGN: Prospective cross-sectional study. SETTING: Department of anaesthesiology at one university hospital. PATIENTS: 26907 consecutive anaesthetic procedures in patients over 15 years of age and a complete preoperative evaluation. Patients undergoing cardiac and obstetric surgery were excluded. MEASUREMENTS AND MAIN RESULTS: Demographic data, preoperative health status, type of anaesthesia, operative procedures, and perioperative incidents (standardised on a national basis) were acquired by means of a computerised anaesthetic record system. Occurrence of at least one perioperative event with impact on postanaesthetic care was computed by a multivariate logistic regression model against 17 variables with different characteristics representing possible risk factors. Fourteen variables proved to be independent risk factors. The weighting of the variables was expressed in scores which added up to form a simple index for each patient. Patients without major risk factors (0-10 points) had a 0.3% risk of suffering from a relevant incident. Patients with more than 60 points had a 28.6% risk. The results were well demonstrated by cross-validation. CONCLUSIONS: The index seems to reflect the risk of relevant perioperative incidents. It can be used for audit purposes. In daily routine, the index could focus our attention on patients with increased perioperative risk. However, it is limited in detecting particular constellations of factors which interact on each other with regard to perioperative risk. PMID- 9754619 TI - Association studies in the presence of comorbidity: design and analysis. AB - Methods for the design and analysis of allelic association studies in the presence of two comorbid disorders are discussed. These methods are applicable to population-based (i.e., case-control) designs. We first develop probability models that represent pathways to the comorbidity of two disorders when it is hypothesized that at least one of the two disorders is associated with a specific allele. These potential pathways are illustrated with the specific example of the association of the human leptin (OB) gene with obesity and major depressive disorders in humans. We then discuss methods of design and analysis using the well-known methods of log-linear analysis [Bishop et al., 1975: "Discrete Multivariate Analysis." M.I.T. Press, Cambridge, MA] to differentiate between these pathways. With the increasing focus in psychiatric genetics on both association studies and pathways to comorbidity we anticipate that these methods will have wide applications. PMID- 9754621 TI - A genome-wide search for schizophrenia susceptibility genes. AB - We completed a systematic genome-wide search for evidence of loci linked to schizophrenia using a collection of 70 pedigrees containing multiple affected individuals according to three phenotype classifications: schizophrenia only (48 pedigrees; 70 sib-pairs); schizophrenia plus schizoaffective disorder (70 pedigrees; 101 sib-pairs); and a broad category consisting of schizophrenia, schizoaffective disorder, paranoid or schizotypal personality disorder, psychosis not otherwise specified (NOS), delusional disorder, and brief reactive psychosis (70 pedigrees; 111 sib-pairs). All 70 families contained at least one individual affected with chronic schizophrenia according to DSM-III-R criteria. Three hundred and thirty-eight markers spanning the genome were typed in all pedigrees for an average resolution of 10.5 cM (range, 0-31 cM) and an average heterozygosity of 74.3% per marker. The data were analyzed using multipoint nonparametric allele-sharing and traditional two-point lod score analyses using dominant and recessive, affecteds-only models. Twelve chromosomes (1, 2, 4, 5, 8, 10, 11, 12, 13, 14, 16, and 22) had at least one region with a nominal P value <0.05, and two of these chromosomes had a nominal P value <0.01 (chromosomes 13 and 16), using allele-sharing tests in GENEHUNTER. Five chromosomes (1, 2, 4, 11, and 13) had at least one marker with a lod score >2.0, allowing for heterogeneity. These regions will be saturated with additional markers and investigated in a new, larger set of families to test for replication. PMID- 9754620 TI - No evidence for linkage of the CHRNA7 gene region in Canadian schizophrenia families. AB - Schizophrenia patients demonstrate a deficiency in the filtering of sensory information, and one specific measure involves a response to the second of a pair of auditory stimuli. A neurophysiological measure of this consists of the electroencephalographic response to pairs of auditory signals, emitted fractions of a second apart. Schizophrenic patients and some of their unaffected relatives show a failure of inhibition of a second tone if it occurs 50 msec after the first. A recent genome scan indicated that the gating defect is linked to the alpha 7 neuronal nicotinic acetyl choline receptor gene on chromosome 15. We genotyped 5 schizophrenia families with a total of 96 subjects with a dinucleotide polymorphic marker located less than 120 kb from the first exon of the alpha 7 neuronal nicotinic acetylcholine receptor gene. Linkage analysis was undertaken using parametric and nonparametric statistical methods. The results of the parametric analysis showed negative lod scores under both narrow and broad diagnosis (lod = -3.6 and -4.8, respectively, at theta = 0), and dominant and recessive modes of transmission of the disease. Nonparametric analysis using GENEHUNTER produced nonsignificant NPL scores (NPL = -0.4 and -0.3 for broad and narrow diagnoses, respectively). In summary, we did not find any evidence that the alpha 7 neuronal nicotinic acetylcholine receptor gene (CHRNA7) is linked to schizophrenia. However, we have not been able to assess the P50 measures in these families. PMID- 9754622 TI - Testing a genetic structure of blood-injury-injection fears. AB - Multivariate genetic analyses were used to examine the genetic and environmental contributions to individual differences in fears of blood, injury, and injections in 659 twin pairs who completed questions concerning fear and fainting around blood, injury, and injections, and fainting in situations not involving blood, as well as the personality scales of Neuroticism, and Harm Avoidance. There was significant familial aggregation of blood fears but univariate analyses were unable to distinguish between additive genetic or shared environmental variables, or both, as the cause. The same was true of blood fainting. Non-blood-injury fainting was best explained by a model assuming shared and unique environmental variables. However, multivariate genetic analyses, which capitalise on extra information contained by all the covariance terms, indicated that the variance in blood-injury-injection fear was principally attributable to unique environmental events specific to this fear and additive genetic factors shared with fainting. The data are discussed in the context of models of blood-injury phobia that identify the need to consider separate etiological mechanisms for fear and fainting. PMID- 9754623 TI - No association of a functional polymorphism in the dopamine D2 receptor promoter region with bipolar or unipolar affective disorders. AB - The dopaminergic system, along with the serotonergic and noradrenergic systems, has been implicated in the etiology of mood disorders. An association study of a functional variant in the promoter region of the dopamine D2 receptor (DRD2) with bipolar affective disorder I or unipolar major affective disorders was performed. Variable expression of the DRD2 gene in vitro has been shown with this promoter polymorphism. One hundred and thirty-one unrelated bipolar patients, 128 unrelated unipolar patients, and 262 controls were used in the study. There were no significant differences in DRD2 allele or genotype frequencies between the affective disorder and control groups. These results do not support a major role for the DRD2 gene in the etiology of either bipolar or unipolar affective disorders. PMID- 9754624 TI - Systematic search for variations in the tyrosine hydroxylase gene and their associations with schizophrenia, affective disorders, and alcoholism. AB - Tyrosine hydroxylase is the rate-limiting step in the biosynthesis of catecholamines. To find variants in the tyrosine hydroxylase (TH) gene that are associated with schizophrenia, mood disorders, or alcohol dependence, all of the exons, the exon-intron boundaries, and the 5' promoter region of the TH gene were systematically screened for variants by single-strand conformation polymorphism analysis followed by direct nucleotide sequencing. Source DNAs for sequencing were from 88 Japanese patients comprised of 17 schizophrenics, 21 with mood disorders, and 50 alcoholics. Two novel variants, T-229A and Val468Met, were identified. Case-control comparisons demonstrated that distribution of these two variants were similar in the controls and the three psychiatric groups. Distributions of the previously reported Val81Met polymorphism alleles and the intron 1 TCAT repeat polymorphism alleles were similar in the four subject groups. Our study indicates that the TH gene is not likely to play a major role in the genetic predisposition to schizophrenia, mood disorders, or alcohol dependence. PMID- 9754625 TI - Patterns of parental transmission and familial aggregation models in bipolar affective disorder. AB - Two recent studies [McMahon et al., 1995: Am J Hum Genet 56:1277-1286; Gershon et al., 1996: Am J Med Genet (Neuropsychiatr Genet) 67:202-207] reported an excess of maternal transmission in bipolar affective disorder in multiply affected families. In a sample of 130 families ascertained through a bipolar proband without regard to psychiatric family history we analysed the frequency of maternal (MAT) and paternal (PAT) transmissions, the morbid risk (MR) in relatives of transmitting mothers and fathers and the inheritance patterns in families with MAT vs. PAT transmission of the disease. In the total sample of 130 families we identified 39 families where the disease was transmitted from the paternal side (PAT families) and 35 families where the disease was transmitted from the maternal side (MAT families). Counting PAT and MAT transmissions in these unilineal families we found nearly equal numbers for both transmission types under a narrow (BP: bipolar disorder, schizoaffective-bipolar type disorder) and a broad definition (AFF: BP, recurrent unipolar depression) of the phenotype. The MRs for narrow and broad phenotypes were not significantly different in any type of PAT relative in PAT families vs. MAT relatives in MAT families. However, in PAT families there were two times more affected females than males with both disease models, while in MAT families there was no MR difference by relatives' sex. The transmission of BP was compatible with the Mendelian major gene model in PAT families and with the multifactorial model in MAT families. Extension of the relatives' phenotype led to borderline non Mendelian major effects in PAT families and reproduced the multifactorial model in MAT families. PMID- 9754626 TI - Variants in the alpha2A AR adrenergic receptor gene in psychiatric patients. AB - In various studies of psychiatric patients, alterations in adrenergic receptor (AR) expression or function have been suggested. Herein, the alpha2A AR gene was screened in 206 patients with schizophrenia, attention deficit hyperactivity disorder (ADHD), autism, alcohol dependence, or cocaine dependence. The entire coding region was examined for single base pair changes, using restriction endonuclease fingerprinting (REF), a screening method that can detect virtually 100% of mutations in 2-kb DNA segments. In the approximately 600 kb of screened sequence, six novel nucleotide changes were identified. The changes resulted in four missense changes (A25G, N251K, R368L, and K370N), and a sequence in the 3' untranslated region. In addition, a silent change (G363G) was found at high frequency in Asians and Native Americans. Of the four missense changes, two found in patients with alcohol/drug dependence occur in highly conserved amino acids, suggesting that these are of likely functional significance. As the alpha2A ARs are widely distributed both pre- and postsynaptically, and as many pharmacological agents with multiple effects target these receptors, the novel missense changes described herein may be candidates for involvement in alcohol/drug dependence, in other clinical disorders or traits, or in differential response to pharmacotherapy. PMID- 9754627 TI - Association analysis in an evolutionary context: cladistic analysis of the DRD2 locus to test for association with alcoholism. AB - Previous studies testing the dopamine D2 receptor (DRD2) locus for association with alcoholism have produced conflicting results. Failure to screen controls for substance abuse and failure to assess alcoholics for severity have been proposed as causes for the inability of some studies to detect an association. We have reevaluated the involvement of DRD2 mutations in susceptibility to alcoholism with a cladistics-based association analysis after restricting an alcoholic sample to more severe cases. For the present study we tested 55 alcoholic probands and 80 normal controls for differences in the frequency of six haplotypes at the DRD2 locus. The haplotypes were derived from five di-allelic polymorphisms spanning all but the first exon of the DRD2 gene. A cladogram constructed from the haplotypes provided the evolutionary context for a nested statistical analysis. We found no significant evidence for association of the DRD2 haplotypes analyzed with the more severe alcoholic phenotype. PMID- 9754628 TI - Evidence for linkage to psychosis and cerebral asymmetry (relative hand skill) on the X chromosome. AB - The hypothesis that psychosis arises as a part of the genetic diversity associated with the evolution of language generates the prediction that illness will be linked to a gene determining cerebral asymmetry, which, from the evidence of sex chromosome aneuploidies, is present in homologous form on the X and Y chromosomes. We investigated evidence of linkage to markers on the X chromosome in 1) 178 families multiply affected with schizophrenia or schizoaffective disorder with a series of 16 markers spanning the centromere (study 1), and 2) 180 pairs of left-handed brothers with 14 markers spanning the whole chromosome (study 2). In study 1, excess allele-sharing was observed in brother-brother pairs (but not brother-sister or a small sample of sister-sister pairs) over a region of approximately 20 cM, with a maximum LOD score of 1.5 at DXS991. In study 2, an association between allele-sharing and degree of left-handedness was observed extending over approximately 60 cM, with a maximum lod score of 2.8 at DXS990 (approximately 20 cM from DXS991). Within the overlap of allele-sharing is located a block in Xq21 that transposed to the Y chromosome in recent hominid evolution and is now represented as two segments on Yp. In one of two XX males with psychosis we found that the breakpoint on the Y is located within the distal region of homology to the block in Xq21. These findings are consistent with the hypothesis that an X-Y homologous determinant of cerebral asymmetry carries the variation that contributes to the predisposition to psychotic illness. PMID- 9754629 TI - Autistic symptoms among children and young adults with isodicentric chromosome 15. AB - A standardized assessment of autistic symptomatology was completed for 29 children and young adults with a supernumerary isodicentric chromosome 15 (formerly known as inverted duplication 15). Although there was variability in severity, 20 individuals with an isodicentric chromosome 15 [idic(15)] had a high probability of being autistic. Eight of the 9 remaining children were under age 5 years and were more sociable than the rest of the cohort. Group characteristics such as gender and seizure presence could not explain the observed difference between older and younger individuals in our study. The natural history of isodicentric 15 syndrome remains to be shown through longitudinal work and may include an age-related risk for developing autism. PMID- 9754630 TI - Novel mutations in the promoter and coding region of the human 5-HT1A receptor gene and association analysis in schizophrenia. AB - Dysfunction of serotonin systems has been implicated in schizophrenia. In the present study, the human 5-HT1A receptor gene containing the 5' untranslated region was screened in order to detect genetic variations, through which alteration of protein function or level of expression might contribute to schizophrenia. Genomic DNAs were isolated from whole-blood samples of 61 unrelated schizophrenic patients and 100 healthy controls. Genetic variations were screened systematically by single-strand conformational polymorphism (SSCP) analysis, followed by direct sequencing of polymerase chain reaction (PCR) product as well as restriction fragment-length polymorphism (RFLP). The novel mutations (-51T --> C, -152C --> G, -321G --> C, -480delA, and -581C --> A) were found in the 5' untranslated region. Furthermore, we found a novel missense mutation (Gly272Asp) in the coding region in addition to the mutations (Pro16Leu, 294G --> A, and 549C --> T) reported previously. No significant differences in genotype frequencies as well as allele frequencies were found between patients and controls. Our data provided no evidence of association between schizophrenia and the variants in the 5' untranslated region as well as the coding region of the human 5-HT1A receptor gene. PMID- 9754631 TI - Variation at the MJD locus in the major psychoses. AB - Expansion of triplet repeats has been seen to underlie several disorders that manifest anticipation. Clinical evidence suggests that anticipation occurs in the major psychoses. We studied the distribution of repeat sizes at the Machado Joseph disease (MJD) locus in a group of patients with the major psychoses. We did not find any large expansions, though 2 patients had alleles that were two repeats larger than in our controls. The difference in allele sizes was larger in the patient sample as compared to the controls. The effect of such large differences might be of functional significance. PMID- 9754632 TI - Linkage and association analysis of chromosomal regions containing genes related to neuroendocrine or serotonin function in families with early-onset, recurrent major depression. AB - Recurrent unipolar depression with an early age of onset is a severe form of unipolar depression that has both genetic and environmental components. We genotyped the members of 16 families identified by probands with early onset (< or = 25 years), recurrent unipolar, major depression for 38 simple sequence tandem repeat polymorphisms (SSTRPs) from chromosomal regions containing 12 genes involved in neuroendocrine or serotonergic functioning. Pairwise linkage analysis was performed with the software package FASTLINK. The affected phenotype was defined four ways, and both dominant and recessive models of depression were analyzed. Seven SSTRPs showed lod scores > 1.00 at theta values between 0.10 0.20. The members of an additional 18 families were genotyped for these seven SSTRPs, and the complete sample of 34 families was evaluated using lod score analysis, affected pedigree member linkage analysis, and within-family association analysis. Evidence for linkage between D11S929 and affective illness remained positive, necessitating the analysis of four additional SSTRPs within 3 cM of D11S929. After all confirmatory analyses were completed, no evidence suggestive of linkage remained between any of the 38 SSTRPs and the affected phenotypes. PMID- 9754633 TI - Quantitative changes in pharmacodynamic parameters of noradrenaline in different rat aorta preparations: influence of endogenous EDRF. AB - 1. The aim of the present study was to assess the role of endothelial cells in the modulation of vasocontractile responses to noradrenaline in rat isolated aorta when cut as standard helical strips or as ring segments. 2. Noradrenaline potency in helical strip preparations evaluated as -logEC50 was greater than that obtained in endothelium-intact ring preparations (9.45 +/- 0.28 versus 8.69 +/- 0.09, respectively) (P < 0.05). The maximum contractile response of helical strips was significantly higher than the response of ring preparations (P < 0.05). 3. Subsequent experiments were performed on helical strips and ring preparations where the endothelium was removed by rubbing the luminal surface of the aorta with filter paper. Removal of the endothelium potentiated the noradrenaline-induced contraction in ring preparations, but not in the helical strips. 4. The nitric oxide synthase inhibitors L-NAME (3 x 10(-5)-3 x 10(-4) M) or L-NNA (1 x 10(4)-3 x 10(-4) M) which were added to the tissue bath potentiated the noradrenaline-induced contraction in the endothelium-intact ring preparations, although only L-NNA induced a statistically significant potentiation. Both L-NAME and L-NNA had no effect on the noradrenaline contraction induced in rings without endothelium, or in helical strips with or without endothelium. 5. Vascular acetylcholine-induced relaxation is dependent on endothelium derived relaxing factor (nitric oxide). Acetylcholine (10(-9)-10(-6) M) induced a concentration-dependent relaxation in noradrenaline preconstricted intact rings. The relaxant response was strongly reduced by L-NAME (3 x 10(-5)-1 x 10(-4) M). The relaxant response to acetylcholine was very weak in ring and helical strip preparations without endothelium, but also, surprisingly, in unrubbed standard helical strips. 6. The present results suggest that the endothelium of standard helical strip preparations may be greatly damaged, a view confirmed by morphological studies. The structural and functional damage of the endothelium induced very important changes in pharmacodynamic parameters such as in the potency and the maximal responses of vascular preparations to noradrenaline. Therefore, caution must be observed when the potency and intrinsic activity of agonists evaluated on different preparations are compared, even if these come from the same vascular segment. PMID- 9754634 TI - Tachykinins in propranolol-augmented, hyperpnoea-induced bronchoconstriction in Taida guinea-pigs: effects of dimethylthiourea. AB - 1. The present authors recently found that a marked hyperpnoea-induced bronchoconstriction (HIB) only occurred in guinea-pigs after treatment with propranolol, a non-selective beta-adrenoceptor antagonist. This study investigated tachykinin-dependent and antioxidant-modulated mechanisms for this propranolol-augmented HIB. 2. Guinea-pigs were pre-treated with an antioxidant, dimethylthiourea (DMTU), or saline for 3 days. On the day of study, each animal was given a dose of propranolol (0.5 mg kg(-1)), then the airway function was examined in the anaesthetized-paralysed animal before, during and after hyperpnoea with a dry 95% O2:5% CO2 gas mixture. Tracheal neutral endopeptidase (NEP) activity and plasma substance P (SP) level were measured after functional study. 3. In the presence of propranolol, HIB was augmented, and was found to be associated with decreased NEP activity and an increased plasma SP level. The augmented HIB was attenuated by DMTU. 4. Therefore, the present results suggest that propranolol-augmented HIB is tachykinin-dependent and is modulated by DMTU. PMID- 9754635 TI - Differential effects of adrenaline and noradrenaline on the hepatic expression of immediate early genes in mice. AB - 1. The effects of intraperitoneally (i.p.) administered noradrenaline and adrenaline on the hepatic expression of immediate early genes (IEGs) were studied in mice. 2. Intraperitoneal injections of various doses (0.2-2 mg kg(-1)) of noradrenaline and adrenaline dose-dependently induced hepatic c-fos and c-jun mRNA levels. The time-course study showed that there was an increase in c-fos and c-jun mRNA levels within 15 min, which reached a peak at 30 min, and returned to the basal levels 1-2 h after noradrenaline or adrenaline injection (2 mg kg(-1), i.p.). A Western blot assay revealed that c-Jun protein levels were maximally increased at 30 min and 1-2 h in noradrenaline- and adrenaline-treated mice, respectively. There was a slight increase in c-Fos protein, while 46-kDa Fra protein was prominently increased. Noradrenaline (2 mg kg(-1), i.p.) induced 46 kDa Fra within 15 min, which reached a maximum at 30 min and returned to the basal levels by 1 h. Adrenaline (2 mg kg(-1), i.p.) induced 46-kDa Fra at 30 min, which returned to the basal levels at 4 h. 3. Noradrenaline (2 mg kg(-1), i.p.) induced increases in c-fos and c-jun mRNA expressions were inhibited by the pre treatment with prazosin (alpha1-adrenergic antagonist; 0.5 mg kg(-1), i.p.), but not with yohimbine (alpha2-adrenoceptor antagonist; 1 mg kg(-1), i.p.) nor with propranolol (beta-adrenoceptor antagonist; 10 mg kg(-1), i.p.). Adrenaline (2 mg kg(-1), i.p.)-induced increases in c-fos and c-jun mRNA expressions were inhibited by the pre-treatment with prazosin or with propranolol, but not with yohimbine. Administration of ICI-118,551 (beta2-adrenoceptor antagonist; 2 mg kg( 1), i.p.), but not betaxolol (beta1-adrenoceptor antagonist; 2 mg kg(-1), i.p.), blocked adrenaline (2 mg kg(-1), i.p.)-induced increases in c-fos and c-jun mRNA expressions. 4. The results suggest that noradrenaline elicits the hepatic c-fos and c-jun mRNA responses by stimulating alpha1-adrenergic receptors, whereas in the case of adrenaline, this is elicited by stimulating both alpha1- and beta2 adrenergic receptors in mice. These catecholamine-induced hepatic IEG responses may be responsible for mediating some of the catecholamine actions in the liver. PMID- 9754636 TI - Electromechanical coupling in human vas deferens: effects of agents that modulate intracellular release of calcium. AB - 1. The effects of ryanodine, cyclopiazonic acid (CPA) and caffeine on electromechanical coupling in human vas deferens were investigated. 2. High [K+]o (120 mM) evoked nifedipine-sensitive contractions of longitudinal and circular muscle which consisted of initial and secondary components. 3. Exposures to ryanodine (< or =10 microM) or CPA (< or = 3 microM) induced a change of basal tension, and higher doses (30 microM) induced intermittent rhythmic contractions of both muscle types in the quiescent tissue. In the presence of the drugs, contraction to high [K+]o was preceded by marked rhythmic activity. 4. In circular muscle, ryanodine (1-30 microM) or CPA (1-30 microM) reduced both components of contractions to high [K+]o. In longitudinal muscle, the drugs enhanced the initial component and prolonged the secondary component. High doses (> or = 10 microM) produced variable effects on the initial component. 5. Caffeine (20 mM) reliably contracted longitudinal, but not circular muscle. Pre exposures to caffeine enhanced both components in the post-caffeine contractions of circular muscle to high [K+]o. In longitudinal muscle, only the initial component (post-caffeine) was enhanced. 6. Contractions evoked in longitudinal muscle by caffeine were not blocked by ryanodine (30 microM) or CPA (30 microM). However, the enhancement of post-caffeine contractions to high [K+]o was inhibited. 7. These results show that ryanodine and CPA produced comparable effects on the excitability of longitudinal and circular muscle in the quiescent tissue, but electromechanical coupling was affected differently. The findings suggest that the muscle types utilize different mechanisms to regulate elevations in cytosolic Ca2+ during stimulation. 8. Electromechanical coupling in both muscle types involves Ca2+ influx via nifedipine-sensitive voltage-operated calcium channels and activation of ryanodine-sensitive calcium-induced calcium release from the sarcoplasmic reticulum (SR). In longitudinal muscle, the SR also buffers increases in cytosolic Ca2+ via a pharmacologically distinct Ca2+ compartment (caffeine releasable but ryanodine/CPA-insensitive). In circular muscle, the SR (ryanodine/CPA-sensitive) serves mainly in the regulation of excitability of the quiescent tissue. PMID- 9754637 TI - Ryanodine- and cyclopiazonic acid-sensitive components in human vas deferens contractions to noradrenaline. AB - 1. The role of calcium stores in noradrenaline- (NA) and caffeine-induced contractions of human vas deferens were investigated using ryanodine and cyclopiazonic acid (CPA) in the presence of the calcium antagonist, nifedipine (1 microM) or in calcium-free/EGTA (1 mM) medium. 2. In either media, NA (100 microM) evoked biphasic contractions of longitudinal muscle and tonic circular muscle contractions. Caffeine (20 mM) evoked longitudinal but not circular muscle contractions. 3. Ryanodine (1-30 microM) or CPA (1-30 microM) inhibited contractions of circular muscle, and the initial but not secondary component of longitudinal muscle contraction to NA. 4. In the presence of nifedipine, pre exposure to caffeine caused a potentiation of circular muscle, and the initial but not secondary longitudinal muscle contractions to NA. The presence of ryanodine or CPA during the caffeine pre-exposures effectively blocked the potentiation of the initial component and reduced the secondary component of the subsequent responses to NA in longitudinal muscle. 5. In calcium-free media, caffeine pre-exposures had little effect on subsequent NA-induced contractions in circular muscle, but reduced both components in longitudinal muscle. The presence of ryanodine or CPA during caffeine pre-exposures produced no further effects on either component of the subsequent NA-induced contraction in longitudinal muscle. 6 In the presence of nifedipine or in calcium-free media, repeated applications of caffeine evoked contractions in longitudinal muscle which were not blocked by either ryanodine or CPA. 7. These results suggest that circular muscle contraction by NA and the initial component of longitudinal muscle to NA both utilize an intracellular pool of calcium that is triggered via a ryanodine sensitive mechanism and replenished via a CPA-sensitive Ca2+-ATPase. 8. In longitudinal muscle, both the secondary component of its response to NA and contraction to caffeine appear to involve an unusual but pharmacologically distinct (ryanodine- and CPA-insensitive) pathway. 9. The quiescence to caffeine of circular muscle may be caused by a relative absence of the ryanodine- and CPA insensitive pathway. PMID- 9754638 TI - Cardiovascular responses evoked by carbachol microinjection into the posterior hypothalamus involves ganglionic nicotinic and muscarinic mechanisms. AB - 1. Microinjection of the cholinergic agonist carbachol (3.3, 5.5 and 13.2 nmol) into the posterior hypothalamic nucleus of conscious rats evokes a dose-dependent increase in blood pressure. The pressor response evoked by the lower doses of carbachol was attenuated by pretreatment with the ganglionic nicotinic receptor antagonist pentolinium (10 mg kg(-1), i.v.) while blockade of V1-vasopressin receptors with [d(CH2)5Tyr(Me)]AVP (20 microg kg(-1), i.v.) reduced the pressor response evoked by the highest dose. 2. The combination of pentolinium and the muscarinic receptor antagonist methylatropine (2 mg kg(-1), i.v.) completely blocked the response evoked by the lower doses while the addition of [d(CH2)5Tyr(Me)]AVP to these two antagonists was required for further inhibition of the pressor response to the highest dose of carbachol. Bilateral adrenal demedullation did not affect the pressor response evoked by 5.5 or 13.2 nmol of carbachol. 3. Treatment of intact and adrenal demedullated rats with pentolinium after the pressor response to 13.2 nmol of carbachol was underway reversed the pressor response, but not to the same degree as that provided by the combination of pentolinium and methylatropine, or pentolinium and [d(CH2)5Tyr(Me)]AVP. 4. Methylatropine or [d(CH2)5Tyr(Me)]AVP caused a slight reversal of the carbachol induced pressor response once it was underway in intact rats. Methylatropine given before or after pentolinium worked with the pentolinium to completely reverse the response. Methylatropine given alone reversed the bradycardia evoked by carbachol to a tachycardia which itself was antagonized by subsequent treatment with pentolinium. 5. These results suggest that the pressor response evoked by carbachol microinjection into the posterior hypothalamic nucleus of conscious rats involves sympathoexcitation and vasopressin release. The sympathoexcitation involves nicotinic and muscarinic receptors in autonomic ganglia. PMID- 9754639 TI - Unique effect of tetrapentylammonium ions on sympathetic transmission in rat vas deferens. AB - 1. Effects of tetrapentylammonium ions (TPA+) on contractile responses induced by electric field stimulation and by exogenous agonist such as noradrenaline, ATP and high K+ were examined in the isolated epididymal half of the rat vas deferens. 2. The electrically evoked monophasic contractions (0.5 Hz, 1 ms pulse duration, 40 V) were potentiated by TPA+ at low concentrations (1-3 microM) and inhibited by TPA+ at a concentration greater than 10 microM. 3. TPA+ (0.3-3 microM) potentiated the contractile responses induced by noradrenaline and ATP, whilst, TPA+ (10 microM) concentration-dependently reduced the agonist-induced contractions. TEA+ enhanced both electrically evoked and agonist-induced contractions. 4. TPA+ (0.3 microM) potentiated 30 mM K+-induced phasic contraction which was inhibited by pretreatment with alpha,beta-methylene ATP (3 microM) and prazosin (3 microM). 5. TPA+ (10-300 microM) reduced the contractions induced by 100 mM extracellular K+ while TBA+ and TEA+ had no effect. 6. The present results show that TPA+ at low concentrations may act at prejunctional nerve membranes to enhance release of contractile transmitters and act as a putative K+ channel blocker at postjunctional membranes to increase muscle contractility in rat vas deferens; whilst at high concentrations TPA+ mainly acts on smooth muscle probably as a non-selective relaxant against the agonist-induced contraction probably through inhibition of Ca2+ influx; this inhibitory effect appears unique for TPA+ since TEA+ and TBA+ did not induce muscle relaxation. PMID- 9754640 TI - cagA-positive Helicobacter pylori and risk for developing gastric carcinoma in Brazil. AB - To evaluate a possible association between infection with cag A-positive strains and gastric carcinoma increased risk we studied 119 Helicobacter pylori-positive patients with gastric carcinoma and 119 matched controls. The presence of cag A gene was investigated by PCR in H. pylori isolates and in gastric biopsy specimens. A significant association was found between cag A-positive status and distal gastric carcinoma for both the intestinal and diffuse types of tumor for both males and females. On the other hand, no association was observed between cag A-positive status and proximal gastric carcinoma. PMID- 9754641 TI - Perinatal correlates of specific histological types of testicular cancer in patients below 35 years of age: a case-cohort study based on midwives' records in Denmark. AB - Our aim was to estimate the risk of specific histological types of testicular cancer associated with perinatal factors. We conducted a case-control study with 357 cases and 704 controls born between 1950 and 1968. Cases were identified through a patient registry of all patients treated in Denmark. Data on parents' age, mothers' parity, birth weight, gestational age and undescended testes were extracted from midwives' records written shortly after delivery. The risk of testicular cancer was highest for those who has the oldest mothers (30-45 years) and for those who had the lowest parity (primipara). For newborns with 2 or more signs of prematurity (born before 8 months of gestation, birth weight below 2,500 g, birth length 40-49 cm, soft nails and undescended testis) the OR was 1.62. The association between undescended testis at birth and testicular cancer was not present for boys born after 1955. The high risk of testicular cancer among primipara was mainly seen for seminomas. All cancer types were associated with mother's age, but only the OR for teratomas was statistically significant. PMID- 9754642 TI - Trends in the incidence of non-melanocytic skin cancer (NMSC) treated in Australia 1985-1995: are primary prevention programs starting to have an effect? AB - Non-melanocytic skin cancer (NMSC) is the most common cancer in Australia, but data on its incidence are not routinely collected by cancer registries. National surveys were conducted in 1985, 1990 and 1995 to estimate NMSC incidence. Trends in incidence between 1985 and 1995 have been examined to determine the impact of primary prevention campaigns aimed at controlling skin cancer in Australia. National random household surveys of Australians aged over 13 years were used to estimate NMSC incidence in 1985, 1990 and 1995. Age- and sex-specific rates by survey year were modelled using Poisson regression. Basal cell carcinoma (BCC) rates in 1995 were 788 per 100,000, an increase of 19% since 1985. Squamous cell carcinoma (SCC) rates rose by 93% over the same period, from 166 to 321 per 100,000. The ratio of BCC:SCC changed from 4:1 in 1985 to 2.5:1 in 1995. BCC rates in latitudes <29 degrees S remained at about 3 times those in latitudes >37 degrees S over the decade. The ratio of SCC incidence between these latitudes changed from around 7:1 to 3:1 over the same period. Although NMSC incidence rates continue to rise, there have been reductions in BCC observed in younger age groups. Incidence rates of NMSC continue to rise in Australia, but there is evidence of a reduction in BCC incidence in younger cohorts. This is evidence that public health campaigns to reduce sun exposure may be having a beneficial effect on skin cancer rates. PMID- 9754643 TI - Proliferative activity of cancer cells in front and center areas of carcinoma in situ and invasive sites of esophageal squamous-cell carcinoma. AB - Intraepithelial carcinoma contiguous with invasive squamous-cell carcinoma is a conspicuous feature of esophageal cancer. However, whether the mechanism of intraepithelial spreading is due to cell proliferation or field carcinogenesis has yet to be clarified. This study investigated the mechanism of intraepithelial spreading by measuring the cell proliferative activity using argyrophilic nucleolar organizer region (AgNOR) and proliferating cell nuclear antigen (PCNA) positive cell counting. We examined the AgNOR number and PCNA-positive ratio (PCNA ratio) in the center and outer edge of intraepithelial carcinoma and in the center and deep margin of invasive squamous-cell carcinoma of the esophagus in 50 specimens from 18 cases of esophageal squamous-cell carcinoma concomitant with contiguous intraepithelial carcinoma. The proliferative activity was thus found to differ between the normal epithelium and cancerous lesions (p < 0.001), between intraepithelial carcinoma and invasive cancer (p < 0.001) and between deep margin and center areas of invasive cancer (p < 0.005). On the other hand, such activity was observed to be similar in the center and outer edge of the intraepithelial spread. These findings suggest that cell proliferation is the main mechanism of tumor progression at the invasive site of cancer, whereas in intraepithelial carcinomatous areas, "field carcinogenesis" or a paracrine mechanism, and not cell proliferation, is thought to be the cause of intraepithelial spread of esophageal cancer. These results therefore support the concept of field carcinogenesis. PMID- 9754644 TI - Loss of heterozygosity on chromosome 14 in nasopharyngeal carcinoma. AB - The main objective of this study was to determine the precise frequency of chromosome 14q loss of heterozygosity in nasopharyngeal carcinomas and to define its minimal deletion regions. Thirty-nine tumors were selected for PCR-based deletion mapping using 19 microsatellite polymorphic markers spanning the long arm of this chromosome. Loss of heterozygosity for at least one marker was observed in 29 (74.4%) tumors, while 24 of these tumors displayed partial loss and provided an informative basis for detailed deletion mapping. Three minimal regions of loss were delineated, the first defined by markers D14S278 and D14S288, the second being between D14S51 and the telomere. These data confirmed 2 potential tumor-suppressor-gene loci at 14q12-13 and 14q32. Interestingly, the third region of loss was located at the T-cell-receptor delta-chain locus. This may reflect another tumor-suppressor-gene locus at 14q11.2, or may be the consequence of a specific genomic rearrangement of this region. In addition, these allelic losses occurred with high frequency in all tumor grades and stages and in all histological sub-types. These findings suggest that the genetic alteration of chromosome 14 is common and crucial during nasopharyngeal-carcinoma development. PMID- 9754645 TI - Risk of colon cancer associated with a family history of cancer or colorectal polyps: the diet, activity, and reproduction in colon cancer study. AB - The Diet, Activity, and Reproduction in Colon Cancer (DARCC) study is a large, multi-center case-control study of colon cancer. We examined family histories of cancer among first-degree relatives obtained by computer-assisted in-person interviews from the DARCC to study the impact of family histories of several cancers and colorectal polyps on colon cancer risk. We examined familial cancer risks both by treating a family history of polyps or cancer as a covariate in a logistic regression model, and by comparing cancer or polyp incidence among relatives of cases to incidence among relatives of controls in a proportional hazards model. There were few differences between the odds ratios (OR) or confidence intervals (CI) generated from logistic regression models and the hazard rate ratios (HRR) generated from the proportional hazards models. Overall, the OR of colon cancer among subjects with a family history of colorectal cancer was 1.77. There were only minor differences in risk by sex, age and subsite. A family history of colorectal polyps also increased risk by about the same amount as a family history of colorectal cancer. The increased risk associated with a family history of polyps did not appear to decrease with age. PMID- 9754646 TI - Body size and colorectal-cancer risk. AB - Individuals whose energy intake exceeds expenditure are at increased risk of colorectal cancer. To determine whether body-size measurements at different ages were risk factors for cancer of the colon-rectum, we carried out a hospital-based case-control study in 6 Italian areas, 2 of which were in the South. Interviews were conducted with 1,217 subjects of both genders with incident histologically confirmed cancer of the colon, 726 with cancer of the rectum, and 4,136 controls hospitalized for acute, non-neoplastic, non-digestive conditions. The questionnaire included information on sociodemographic factors, and physical activity, a validated dietary history, height, weight at diagnosis and at 12, 30 and 50 years of age and waist-to-hip ratio (WHR). After allowance for education, physical activity, energy intake, family history of colorectal cancer and recent change in weight, the body-mass index (BMI) was significantly associated with colorectal-cancer-risk in men (odds ratio, OR, in highest vs. lowest quintile = 1.7; 95% confidence interval, CI, 1.3-2.3), but not in women (corresponding OR = 0.9; 95% CI, 0.7-1.2). Cases of both gender tended to have higher BMI than controls in adolescence, young adulthood and middle age. Height appeared unrelated to risk. In women, but not in men, WHR was positively associated with risk, independently of BMI (OR for > or = 0.90 vs. < or = 0.81 = 1.6; 95% CI; 1.2 2.1). Thus, excessive weight predicts colorectal-cancer risk in men, whereas abdominal obesity (i.e., a high WHR) represents a more reliable risk indicator in women. PMID- 9754647 TI - HPV 16 infection and progression of cervical intra-epithelial neoplasia: analysis of HLA polymorphism and HPV 16 E6 sequence variants. AB - High-risk human papillomavirus (HPV) infection plays an important role in cervical intra-epithelial neoplasia (CIN), but HPV infection alone is not sufficient for progression to cervical cancer. Several lines of evidence suggest that cellular immune surveillance is important in the control of HPV infection and the development of CIN. The presentation to T cells of target viral peptides in the context of HLA molecules is influenced by the genetic polymorphisms of both HPV and HLA and thereby influences the host immune response and clinical outcome of HPV infection. HLA class I and II polymorphism in susceptibility for HPV 16 infection, development and progression of CIN was analyzed in a group of 118 patients participating in a prospective study of women with initial abnormal cytology. Patients were stratified according to HPV status and course of the disease. HLA-B*44 frequency was increased in the small group of patients with a lesion that showed clinical progression during follow-up [OR = 9.0 (4.6-17.5), p = 0.007]. HLA-DRB1*07 frequency was increased among HPV 16-positive patients compared with patients who were negative for all HPV types [OR = 5.9 (3.0-11.3), p = 0.02]. Our results are consistent with the immunogenetic factors associated with disease progression being different from those associated with susceptibility to HPV 16 infection. Sequencing of the HPV 16 E6 and E7 open reading frames of a subset of these patients (n = 40) showed the frequency of HPV 16 variants to be similar to other studies. However, there was no significant correlation between variant incidence and disease progression or viral persistence and no significant correlation with any HLA allele. It appears that multiple HLA types can influence HPV 16-associated cervical dysplasia but the role of HPV 16 variants in disease progression and susceptibility in relation to HLA polymorphism remains unclear. PMID- 9754648 TI - Age-specific familial risks in common cancers of the offspring. AB - Quantitative data on familial cancer risks are important for clinical, psychological and scientific reasons. The available estimates carry many uncertainties due to sample size and possible bias in data collection and often refer to first-degree relatives of unspecified age and sex. We calculated sex- and age-specific familial hazard ratios (FHRs) of cancer in offspring aged 15-53 years of cancer probands at 16 male and 17 female cancer sites, based on registered nation-wide data, free from bias. The familial risks in offspring were high, > 5 for thyroid (FHR 10.7 in all offspring, CI 95% 6.9-16.6), and testicular cancer (FHR 5.4, CI 95% 2.6-11.3), or intermediate, FHR 2-5, for colon, rectal, lung, breast, cervical, uterine, ovarian, skin (melanoma and squamous cell) and other endocrine gland cancers. FHRs < 2.0 were observed for stomach, renal and nervous system cancers, lymphomas and leukemias. Some sex differences were observed: FHRs for male breast (only 2 cases) and thyroid cancers were over 2 times higher than the respective female ones. When parents were diagnosed before age 50 years, offspring were at an increased risk of familial breast, renal, skin (melanoma), nervous system, thyroid and non-thyroid endocrine gland cancers, particularly affecting young (< 40 years) individuals. The parental diagnostic age also affected offspring's risk of colon, rectal, uterine and ovarian cancers, but young individuals were not at a particular risk. No effect of age was noted for cervical cancer and lymphoma. PMID- 9754649 TI - P-glycoprotein-mediated methotrexate resistance in CCRF-CEM sublines deficient in methotrexate accumulation due to a point mutation in the reduced folate carrier gene. AB - We have previously described a series of methotrexate (MTX)-selected CCRF-CEM sublines (CEM/MTX R1-3) displaying increased resistance to drugs associated with the multidrug resistance phenotype and have provided evidence that MDR1 P glycoprotein contributes to multifactorial MTX resistance in these cells. We have also suggested that P-glycoprotein-mediated MTX transport arises in these cells due to a deficiency in the normal MTX transport route, the reduced folate carrier (RFC). We have now determined the nucleotide sequence of the RFC gene in CEM/MTX R1-3 cells and confirm that the carrier is defective in these cells as a result of a premature stop mutation at codon 99, which severely truncates the encoded protein. CEM/MTX R3 cells were removed from MTX, and a series of sublines with increasing MDR1 expression were derived, following selection with vincristine. These cells show increasing cross-resistance to vincristine as well as other drugs associated with the multidrug resistance phenotype. More importantly, the increased P-glycoprotein expression correlates with increased resistance to MTX, supporting the hypothesis that in cells with a defective carrier protein, MTX can become a substrate for P-glycoprotein. Our data have implications for the P glycoprotein-mediated transport of other hydrophilic drugs in situations where the relevant carrier protein has been functionally inhibited. PMID- 9754650 TI - Constitutive expression of the Wilms tumor suppressor gene (WT1) in renal cell carcinoma. AB - The expression of the Wilms tumor suppressor gene WT1 is largely restricted to elements of the developing urogenital system. In the fetal kidney, WT1 transcripts are present at low levels in the condensing mesenchyme and at much higher levels in differentiating glomerular epithelium and are not detected in other mesenchymal-derived epithelial structures such as the proximal and distal tubules. However, WT1 expression is observed in tubule-like elements found in some Wilms tumors. As renal cell carcinoma (RCC) of the clear cell type is one of the most prevalent adult tumors of the kidney, and is thought to originate from the epithelial cells of the proximal tubules, we studied WT1 expression in RCCs. Despite the absence of WT1 in normal primary epithelial cells derived from proximal tubules, RCC tumors and tumor-derived cell lines expressed WT1 RNA. Immunocytochemical analyses of tumor cryosections showed widespread expression throughout the poorly differentiated epithelial components of the tumor. Immunoblots of RCC samples detected a normal size WT I protein and reciprocal antibody immunoprecipitations of RCC cell extracts indicated that WT I interacts with p53 as has been demonstrated for normal human fetal kidney. The aberrant expression of functional WT1 in RCC may represent a reversion to a more de differentiated phenotype and may contribute to the tumorigenic phenotype by inappropriately activating or repressing genes involved in growth regulation. PMID- 9754651 TI - Interleukin (IL)-15 induces survival and proliferation of the growth factor dependent acute myeloid leukemia M-07e through the IL-2 receptor beta/gamma. AB - We have analyzed the effects of IL-15, a growth factor with IL-2-like properties produced by dendritic and stromal cells, on 3 GM-CSF/IL-3-dependent AML cell lines: M-07e, UT-7 and TF-1. M-07e cells proliferated in response to IL-15, while UT-7 and TF-1 cells failed to respond. In addition, IL-15 supported long-term proliferation of M-07e cells, thus allowing selection of a subline (M-07SB), which displayed an enhanced sensitivity to IL-15. M-07e and M-07SB cells undergo apoptosis following 48-hr growth factor (GM-CSF or IL-15) starvation, as detected by cytofluorimetric analysis and DNA laddering. IL-15 (20 ng/ml) prevented apoptosis in both cell lines. M-07e and M-07SB expressed IL-2R beta, IL-2R gamma, Jak-1 and Jak-3 mRNA, while IL-15R alpha mRNA was undetectable. In contrast, IL 15R alpha was expressed in UT-7 and TF-1 cells, which lacked expression of IL-2R beta mRNA and, in the case of UT-7, also of Jak-3 mRNA. Accordingly, surface IL 2R beta protein was identified only in M-07e and M-07SB cells, by indirect immunofluorescence, while no expression of IL-2R alpha and IL-15R alpha was detected. Anti-IL-2R beta antibodies (10 microg/ml) efficiently blocked (90% inhibition) the proliferation and the anti-apoptotic effect induced by IL-15, while anti-GM-CSFR alpha antibodies had no effect. Anti-IL-2R gamma antibodies were less efficient at proliferation inhibition but synergized with suboptimal concentrations of anti-IL-2R beta antibodies. Our data suggest a role of IL-15 as an anti-apoptotic and mitogenic growth factor for a subset of myeloid leukemias expressing a functional IL-2R beta/gamma complex. PMID- 9754652 TI - Bone marrow-derived dendritic cells pulsed with a tumor-specific peptide elicit effective anti-tumor immunity against intracranial neoplasms. AB - Although the central nervous system (CNS) is often regarded as an immunologically privileged site, it is well established that specific CNS immunoreactivity can be generated through peripheral vaccination with CNS antigens. Dendritic cells (DC) are potent antigen presenting cells of hematopoietic origin that have emerged as a promising tool for cancer immunotherapy capable of evoking significant anti tumor immunity when pulsed with tumor-associated peptides. To explore a role for DC-based immunization strategies for the treatment of CNS tumors, we developed a brain tumor model using the C3 sarcoma cell line which expresses the tumor specific, major histocompatibility complex (MHC) class I-restricted peptide epitope E7(49-57). Syngeneic C57Bl/6 mice receiving intravenous (i.v.) injections of bone marrow-derived DCs pulsed with E7 peptide were effectively protected against a subsequent intracerebral challenge with C3 tumor cells. More importantly, this systemic immunization strategy was effective in a therapy model as 67% of animals (10 of 15) with established (day 7) intracerebral C3 tumors treated with 3 weekly injections of E7 peptide-pulsed DCs achieved a long-term survival (>90 days) while no control animals survived beyond day 41. In vivo depletion of CD8+ cells, but not CD4+ or asialo-GM1+ cells, abrogated the efficacy of E7 peptide-pulsed DC therapy of established tumors, indicating a pivotal role of specific CD8+ T-cell responses in mediating the anti-tumor effect. Our findings support the hypothesis that effective CNS anti-tumor immunoreactivity can be generated with DC-based tumor vaccines. PMID- 9754653 TI - Identification of HER2/neu-derived peptide epitopes recognized by gastric cancer specific cytotoxic T lymphocytes. AB - We have derived HLA-A2.1-restricted, gastric cancer-specific cytotoxic T lymphocyte (CTL) lines by repetitive in vitro stimulation of tumor-associated lymphocytes (TAL) with autologous tumor cells. The HER2/neu specificity of these gastric cancer-specific CTLs was demonstrated using HER2/neu-transfected cell lines and HER2/neu-expressing tumors, and with a set of HER2/neu-derived peptide epitopes. Gastric cancer-specific CTLs specifically lysed autologous and allogeneic HLA-A2.1+, HER2/neu+ gastric cancer cells, HER2/neu-transfected C1R/A2 cell lines (HLA-A2.1+, HER2+) and HLA-A2.1-transfected SW626 tumor cell lines (HLA-A2.1+, HER2+). This recognition could be inhibited by anti-HLA-A2 antibody or by cold target HER2/neu-transfected C1R/A2 cells. Our results demonstrate that the HER2/neu-encoded HLA-A2.1-associated epitopes recognized by CTLs are presented as naturally processed peptides on gastric cancer lines. Furthermore, 3 of 19 tested HER2/neu-derived peptide epitopes [HER2(9(106)), HER2(9(369)), HER2(9(689))], which all bound HLA-A2.1 with high (IC50 < 50 nM) affinity, were able to sensitize HLA-A2+ C1R/A2 cells to be recognized by the gastric cancer specific CTLs, demonstrating the immunodominance of these epitopes. In conclusion, our findings implicate HER2/neu-derived epitopes as potential candidates for novel immunotherapy and vaccine strategies against gastric cancer. PMID- 9754655 TI - Clonal deletion of thymocytes as a tumor escape mechanism. AB - Clonal deletion of thymocytes is a major event in T-cell tolerance and might represent a tumor escape mechanism. Previously, we have shown that class II restricted, Id-specific, CD4+ T cells in T-cell receptor (TCR)-transgenic mice confer resistance against the MOPC315 plasmacytoma. In this report, we have investigated whether monoclonal immunoglobulin (Ig) produced by a plasmacytoma can induce deletion of thymocytes specific for the variable parts of Ig, i.e., the idiotype (Id). Large numbers of MOPC315 tumor cells were injected s.c. in the TCR-transgenic mice to overwhelm the CD4+ T-cell-mediated protection. When the MOPC315 plasmacytomas reached a weight of approximately 0.5 g (serum myeloma protein M315 about 50 microg/ml), immature CD4+ 8+ and mature CD4+ transgenic thymocytes became progressively deleted. Apoptotic thymocytes were already detectable when tumors were 2 mm in diameter (serum M315: 5 microg/ml, or 0.03 microM). The negative selection was Id-specific, because an Id-negative plasmacytoma failed to induce deletion. Injection of purified MOPC315-myeloma protein (M315) i.p. caused a profound reduction of Id-specific thymocytes. Enriched thymic dendritic cells (DC) from tumor-bearing animals were found to be primed with lambda2(315) and induced apoptosis of thymocytes in vitro. Our results indicate that circulating myeloma protein is processed and presented by thymic antigen-presenting cells (APC), and induces deletion of Id-specific thymocytes. Deletion of tumor-specific thymocytes may represent a tumor escape mechanism in patients with cancers that secrete or shed tumor antigens. The possibility that vaccination with tumor Ig or genes encoding for it may induce tolerance instead of protection should be taken into consideration. PMID- 9754654 TI - Frequency and relative fraction of tumor antigen-specific T cells among lymphocytes from melanoma-invaded lymph nodes. AB - Several tumor antigens have been described as candidates for immunotherapy. Our study compared HLA-A2-restricted epitopes from 5 antigens commonly expressed by melanomas, i.e., Melan-A/MART-1 peptides (26-35 and 27-35), tyrosinase (368-376), gp-100 (280-288), MAGE-3 (271-279) and NA17-A (1-10), for their relative capacity to promote the development of cytotoxic and cytokine-producing specific CD8+ lymphocytes within melanoma-invaded lymph nodes. We used short-term cultured melanoma-invaded lymph node lymphocytes (MILLs) and tested responses developed by these cells to peptide-pulsed TAP-deficient T2 cells. We measured both the lytic response developed by MILLs and the fraction of these cells that secreted interferon-gamma (IFN-gamma), as deduced from intracellular cytokine labeling. Reverse transcription-polymerase chain reaction (RT-PCR) was used to analyze the expression of the 5 antigens within melanoma-invaded lymph nodes. Melan-A/MART-1, tyrosinase and gp-100 peptides were recognized by MILLs derived, respectively, from 8 of 20, 5 of 19 and 4 of 20 melanoma-invaded lymph nodes expressing these antigens. Most MILLs specific for Melan-A/MART-1 and tyrosinase exhibited both lysis and IFN-gamma responses, whereas most of those specific for gp-100 developed only lysis. Weak lysis without IFN-gamma secretion was developed against NA17-A and MAGE-3 peptides by MILLs from, respectively, 3 of 9 and 2 of 14 lymph nodes expressing these antigens. Our data show a prevalence of both cytotoxic and IFN-gamma-secreting effector T cells specific for differentiation antigens within HLA-A2 melanoma-invaded lymph nodes, which makes these antigens attractive targets for specific immunotherapy. PMID- 9754656 TI - Differentiation induction by a tumor-necrosis-factor mutant 471 in human myelogenous leukemic cells via tumor-necrosis-factor receptor-p55. AB - The present study examined differentiation-inducing activity by various tumor necrosis-factor(TNF) mutants against the human leukemic cell lines HL-60 and U 937. Mutant TNF 471, from which 7 N-terminal amino acids of native TNF were deleted and Pro8, Ser9 and Asp10 were replaced by Arg, Lys and Arg, possessed the highest activity among the TNF mutants, and its activity was 120-fold that of native TNF. The various biological activities of TNF were signaled through 2 distinct receptors, p55 and p75. Although cytotoxicity was reported to involve mainly p55, this differentiation-inducing activity was not well understood. The fact that the affinity of TNF 471 was higher to p55 and lower to p75 than that of native TNF by a binding competition assay suggested that the differentiation inducing activity was also signaled through p55. To verify this hypothesis, the human myelogenous leukemic cell line, KG-1, which scarcely expresses either receptor and does not differentiate with TNF, was transduced with the p55 or p75 gene. Subsequently p55 transfectants manifested a greater ability to differentiate; however, p75 transfectants did not differ from parental cells or from mock-transfectants. Further, the differentiation of p55 transfectants induced by TNF was reduced by the inhibitor of protein-kinase-C (PKC), staurosporine. These results indicate that the differentiation-inducing activity was signaled through the TNF receptor, p55, via PKC and that the excellent ability of TNF 471 to induce differentiation was related to its high affinity for p55. PMID- 9754657 TI - Expression of RGD minus fibronectin that does not form extracellular matrix fibrils is sufficient to decrease tumor metastasis. AB - Fibronectin (FN) is a plasma and extracellular matrix (ECM) glycoprotein, the expression of which may modulate the invasive and metastatic abilities of cancer cells. LMM3 is a cell line derived from the highly metastatic mouse mammary adenocarcinoma MM3 and is unable to express FN both at protein and mRNA levels. To study the role of FN in the metastatic process, LMM3 cells were stably transfected with 2 variants (wt and RGD-minus) of a full length human FN cDNA. All analyzed clones secreted recombinant FN and although none assembled FN in the ECM they showed an in vitro reduced migratory ability and an increased adhesive capacity. FN-producing cells were assayed for experimental and spontaneous metastasis. All clones tested showed a significant reduction in the number of experimental lung metastasis when compared with a control clone. Similar trends were observed for spontaneous metastatic ability. Our results indicate that the expression of FN that lacks the well-recognized RGD cell-binding site and that does not form ECM fibrils, is sufficient to decrease the metastatic potential of cancer cells. Our results also suggest that an RGD-independent mechanism may be acting in the prevention of metastasis. PMID- 9754658 TI - Comparison of carcinoembryonic antigen promoter regions isolated from human colorectal carcinoma and normal adjacent mucosa to induce strong tumor-selective gene expression. AB - To establish in vivo gene therapy against cancer, it is requisite to induce strong, cancer cell-selective expression of a therapeutic gene. Comparison of the promoter activity of 5' flanking regions of the carcinoembryonic antigen (CEA) gene isolated from various origins is therefore of considerable interest. The 5' flanking region of the CEA gene between -135 and +69 bp upstream from the transcriptional start site, which is recognized as the core promoter region, was isolated from CEA-producing human colorectal carcinoma (CRC), normal adjacent mucosa, CEA-producing cell lines and CEA-non-producing cell lines. No mutations were observed by single-strand conformation polymorphism in the CEA promoter regions. Subsequent sequence analysis revealed that there were no mutations in the CEA promoter regions isolated from CEA-producing CRC and normal adjacent mucosa. Furthermore, nuclear extracts prepared from CEA-producing human CRC cells could equally bind to both the CEA promoter fragments isolated from CEA-producing CRC and normal mucosa. Both CEA promoter regions could direct 5- to 20-fold higher expression of a luciferase reporter gene in CEA-producing cells than in CEA-non-producing cells. Therefore, we suggest that the use of either CEA promoter region isolated from CRC or normal mucosa is equally effective to induce strong, CEA-producing cancer-selective expression of a therapeutic gene. PMID- 9754659 TI - Inhibition of cancer cell growth by all-trans retinoic acid and its analog N-(4 hydroxyphenyl) retinamide: a possible mechanism of action via regulation of retinoid receptors expression. AB - In order to better understand the mechanisms that underlie the antiproliferative effect of retinoids, we have examined the response of human carcinoma cell lines to all-trans retinoic acid (RA) and N-(4-hydroxyphenyl) retinamide (4HPR) in terms of cell growth, apoptosis and regulation of retinoic acid receptors (RARs) and retinoid X receptors (RXRs) mRNA. GLC82 (lung adenocarcinoma), BGC823 (stomach adenocarcinoma) and EC109 (esophageal squamous carcinoma) cells were treated with 10 microM of RA or 4HPR for various length of time and analyzed. The results show that growth inhibition by RA and 4HPR in GLC82 and BGC823 cells correlates with the induction of RARbeta2 gene, whereas RA resistance in EC109 cells parallels loss of RARbeta2 induction. Exogenous RARbeta2 expression did not restore RA responsiveness in EC109 cells, but potentiated 4HPR-induced growth inhibition, suggesting that 4HPR acts at least in part via the RARbeta receptor. We speculate that the loss of RARbeta2 inducibility in EC109 cells may be due to an unknown repressor. PMID- 9754660 TI - Transfection of the type I TGF-beta receptor restores TGF-beta responsiveness in pancreatic cancer. AB - Transforming growth factor-beta (TGF-beta) signaling is initiated following heterodimerization of the type II TGF-beta receptor (TbetaRII) with the type I TGF-beta receptor (TbetaRI). Both receptors are required for TGF-beta responsiveness. In the present study, we characterized the actions of TGF-beta1 in T3M4 human pancreatic cancer cells, which express low levels of TbetaRI and high levels of TbetaRII. Cells were transiently transfected with p3TP-Lux, a TGF beta-responsive luciferase reporter gene construct. TGF-beta1 was without effect in parental T3M4 cells, but caused a time- and dose-dependent increase in luciferase activity in T3M4 cells co-transfected with a TbetaRI cDNA expression vector. Co-transfection of TbetaRI with a truncated Smad4 cDNA that is known to block TGF-beta-dependent signaling, abrogated the TbetaRI-induced increase in luciferase activity. Sequencing of the TbetaRI and the Smad4 genes in T3M4 cells did not reveal any mutations. These findings indicate that one mechanism for TGF beta resistance in pancreatic cancer is due to a quantitative decrease in TbetaRI expression. PMID- 9754661 TI - Soluble fibronectin promotes migration of oral squamous-cell carcinoma cells. AB - We have shown that a fibronectin (FN) matrix is required for the organization of tenascin-C (TN-C) matrices by peritumor fibroblasts (PTF) cultured from tissue surrounding oral squamous-cell carcinoma (SCC). In the present study, we detected alternatively spliced FN containing both the EDA and EDB domains decorating the reactive stroma adjacent to the invading tumor nests in oral SCC biopsies. In vitro, PTF cells organized an extensive FN matrix rich in the EDA domain and containing a small amount of EDB. In contrast, normal human fibroblasts deposited a FN matrix which expressed only the EDA domain. PTF-conditioned medium (CM), shown to enhance migration of oral SCC cells on TN-C, was found to enhance their migration on FN and invasion of a reconstituted basement membrane. Addition of antibodies to FN to the PTF-CM inhibited SCC-cell migration on TN-C, and depletion of FN from the PTF-CM abolished its ability to induce migration or invasion by oral SCC cells, suggesting that FN promotes the migration and invasion of oral SCC cells. Western blots of the PTF-CM identified FN containing the EDA but not the EDB domain. When soluble FN was added to the control medium in the lower chamber of the Transwell system, SCC-cell migration increased significantly. These results demonstrate that both the EDA and the EDB domains of FN are expressed in the extracellular matrix of oral SCC in vivo and PTF in vitro and indicate that FN is the probable chemotactic factor in the PTF-conditioned medium. PMID- 9754662 TI - Zidovudine to prevent mother-to-infant HIV transmission in developing countries: any questions? PMID- 9754663 TI - Potential implications of the integrated management of childhood illness (IMCI) for hospital referral and pharmaceutical usage in western Uganda. AB - The integrated management of childhood illness approach (IMCI) is currently being implemented by a number of countries worldwide. This is the second report from a study in western Uganda comparing the assessment and classification of disease by medical assistants using the IMCI algorithm with that of hospital-based general medical officers, who used their clinical judgement to assess and provide treatment. Treatment prescribed by the hospital medical officers was compared to that indicated by IMCI disease classifications. The study population comprised 1226 children aged 2-59 months. Medical assistants had some difficulty in completing the IMCI assessment, leading to incorrect classification of findings in 138 of 1086 completed forms (13%). If their classifications had been used to decide on hospital referral, 37 children who met IMCI criteria for referral would have been sent home. Consultations took on average 7.2 min, longer than usual for several African countries. Use of the IMCI guidelines would have referred 16.2% of children to hospital, compared with 22% referred by the medical officers. Use of IMCI could have reduced the cost of medication to US$0.17 per child compared to the treatment cost of US$0.82 as prescribed by medical officers. Medical officers prescribed both a greater number and a greater variety of drugs than indicated by the IMCI algorithm. Compared to the present management of sick children by medical officers at Kabarole district hospital, using the IMCI algorithm would bring major changes in pharmaceutical use and referral practices. However, there is concern about the difficulty medical assistants had in using it, and the potential for longer consultation times. PMID- 9754664 TI - Origin and prevention of airport malaria in France. AB - Since 1969, 63 cases of airport malaria have been reported in Western Europe, 24 of which occurred in France. Most were due to Plasmodium falciparum. In 1994, 7 cases occurred in and around Roissy Charles de Gaulle airport (CDG), showing 4 types of contamination: among employees working on airstrips or opening containers, among residents living near the airport, among people living at some distance from the airport after a secondary transport of vectors, and by vectors transported in luggage. In-flight or stop-over infection is not considered as airport malaria. The infective anophelines originated from airports where malaria transmission occurs, mostly in subsaharan Africa. A tentative list is given taking into account aerial traffic with France. Surveys in the airports of Dakar (Senegal), Cotonou (Benin), Abidjan (Cote d'Ivoire) and Yaounde (Cameroun) found potential vectors in all of these from July to September. After 1994, the Controle Sanitaire aux Frontieres (CSF) in charge at CDG concentrated its efforts on the flights at risk, as well as information and sensitization of airline companies, which resulted in 73% and 87% of the flights at risk being properly disinsected in 1995 and 1996. Despite pyrethroid resistance in Anopheles gambiae s.s. in West Africa, the efficacy of aircraft spraying with permethrin aerosols is still acceptable. However, surveillance of resistance should be improved and search for nonpyrethroid insecticides suitable for aircraft strongly encouraged. PMID- 9754665 TI - Seasonal density, sporozoite rates and entomological inoculation rates of Anopheles gambiae and Anopheles funestus in a high-altitude sugarcane growing zone in Western Kenya. AB - An entomological study was conducted on vectors of malaria and their relative contribution to Plasmodium falciparum transmission in Mumias, a high-altitude site and large-scale sugarcane growing zone in Kakamega district, western Kenya. Anopheles gambiae s.l., the predominant vector species, represented 84% (n=2667) of the total Anopheles mosquitoes collected with An. funestus comprising only 16%. Polymerase chain reaction (PCR) identified all 600 specimens of the An. gambiae complex tested as An. gambiae sensu stricto, an indication that it is the only sibling species represented in the high-altitude sites in western Kenya. Plasmodium falciparum sporozoite rates of 6.3% (133/2118) for An. gambiae s.l. and 9.5% (38/402) for An. funestus by ELISA were obtained in Mumias. None of 1600 mosquitoes tested for P. malariae sporozoites was positive. ELISA tests of mosquito blood meals indicated a high tendency of anthropophagy, a behaviour contributing significantly to malaria transmission by the vector species, with 95.9%, 4.86% and 0.2% having taken at least one blood meal on human, bovine and avian hosts, respectively Malaria transmission intensity was low as revealed by the low entomological inoculation rates (EIR) recorded. The EIR values for An. gambiae s.l. were 29.2 infective bites per person per year (ib/p/year) and 17.5 ib/p/year for An. funestus in Mumias. The highest inoculation rate for both vector species was 7.0 ib/p/month in July. Plasmodium falciparum parasite rate among asymptomatic children was 55.4% and 44% in the wet (July-September) and dry (December-February) seasons, respectively. These results indicate that malaria transmission intensity in the high-altitude site is low but perennial, with transmission being maintained by An. gambiae s.s. and An. funestus. PMID- 9754666 TI - Schistosoma mansoni, intestinal parasites and perceived morbidity indicators in schoolchildren in a rural endemic area of western Cote d'Ivoire. AB - There is a great need for rapid and low-cost identification of communities at high risk of intestinal schistosomiasis. We report the development of a questionnaire approach that may do so. In the first phase, 209 schoolchildren from 3 neighbouring villages in a rural area endemic for intestinal schistosomiasis in western Cote d'Ivoire were screened for Schistosoma mansoni and other helminths on 4 consecutive days using Kato-Katz thick smears. Daily infection prevalences of S. mansoni were high (60%-71%) and the cumulative infection prevalence was 92.3%. Infections with hookworms and Ascaris lumbricoides were also frequent, with cumulative prevalences of 60.8% and 38.3%, respectively. On day 3, the presence of Entamoeba histolytica/E. dispar and Giardia lamblia was assessed by a faecal concentration procedure. In the second phase, focus group discussions (FGD) were conducted: in each village one FGD with heavily infected children and one FGD with lightly or S. mansoni-uninfected schoolchildren to assess their perception of morbidity. The aim was to establish local terms indicating S. mansoni infections. 'Diarrhoea', 'blood in the stools', 'stomach disorders' and 4 terms in the local Yacouba/Dioula languages were frequently used by infected children. A simple questionnaire was then developed and the headteachers interviewed all schoolchildren individually. 'Blood in stools', gnon and toto were reported significantly more frequently among moderately and heavily S. mansoni-infected children than by those not or only lightly infected. The term gloujeu indicated borderline significance. The best diagnostic performance was found for 'blood in stool' (sensitivity: 47%; specificity: 76%; positive predictive value: 66%; negative predictive value: 60%). All schistosomiasis infections were treated with a single oral dose of praziquantel (40 mg/kg body weight) and the same questionnaire was re administered 6 weeks post-treatment. Statistically significantly less children reported having had 'blood in stool' and 'gloujeu' after treatment (McNemar's (chi2-test, P < 0.01). We conclude that 'blood in stool', 'gnon', 'toto' and 'gloujeu' are the most reliable reported symptoms for rapid and low-cost identification of communities that are at high risk of S. mansoni infections in Cote d'Ivoire. PMID- 9754667 TI - Diagnostic significance of Schistosoma mansoni proteins Sm31 and Sm32 in human schistosomiasis in an endemic area in Egypt. AB - We performed a series of ELISAs to evaluate the diagnostic significance of two Schistosoma mansoni proteins, Sm31 (cysteine proteinase, cathepsin B) and Sm32 (asparaginyl endopeptidase). Our study populations were chosen from two villages in an endemic area close to Alexandria. Using fusion proteins MS2-Sm31 and MS2 Sm32 as antigens, 70% and 78.9%, respectively, of patient sera from 134 parasitologically confirmed cases reacted positively. The percentage of seropositivity increased to 84.5% when parasite-derived proteins Sm31 and Sm32 were used. The serum levels of antibodies to these two proteins in recombinant or native forms do not correlate with intensity of infection and hence are detected even when egg counts are low, which makes proteins Sm31 and Sm32 useful antigens in the identification of S. mansoni infected cases, particularly in endemic areas in Egypt. PMID- 9754668 TI - Sonographic prediction of variceal bleeding in patients with liver fibrosis due to Schistosoma mansoni. AB - Several studies have shown that the characteristic hepatic abnormalities induced by Schistosoma mansoni detectable by ultrasound correlate with the degree of oesophageal varices. So far the value of ultrasound for predicting variceal haemorrhage has not been assessed. Fifty Brazilian patients with schistosomal periportal fibrosis from Alagoas State, 18 of whom had already bled from oesophageal varices, were enrolled in a combined cross-sectional and longitudinal study and investigated clinically, by endoscopy and by ultrasound. Twenty-seven of the patients were monitored until another bleeding episode, death or for a minimum of 28 months. Eight of these patients could be followed up for a further three years. A sonographic score, which accounts for the degree of echogenic periportal thickening and of portal vein dilatation, was calculated for all patients. A highly significant correlation (P < 0.0001) existed between the sonographic score and the occurrence of previous variceal haemorrhage, paralleled by a similar correlation between the sonographic score and the degree of oesophageal varices (P < 0.001). In the 27 patients monitored longitudinally, the sonographic score indicated the risk of future variceal bleeding (P < 0.0001). The sonographic score reliably predicts the risk of variceal bleeding in individual patients with periportal fibrosis. Hence, the application of endoscopy, if available at all in endemic areas, may be restricted to the patients at risk of future variceal bleeding, as determined by ultrasound. Since portable devices can be carried even to remote areas, the application of the proposed score in community surveys could provide a new means for the identification of high-risk patients in S. mansoni-infected populations. PMID- 9754669 TI - Megazol combined with suramin: a chemotherapy regimen which reversed the CNS pathology in a model of human African trypanosomiasis in mice. AB - Chemotherapy for human African trypanosomiasis (HAT), or sleeping sickness, is unreliable because of resistance, refraction and toxic and adverse side-effects. Using a long-term experimental model of HAT with involvement of the central nervous system (CNS), we tested the ability of a megazol and suramin combination treatment to eliminate CNS trypanosomes. This consisted of 20 mg suramin per kg body weight administered intraperitoneally (i.p.), followed 24 h later by 4 daily doses (80 mg/kg) of megazol given either i.p. or per os. One week post-treatment, neurological disorders had disappeared. One of 15 mice relapsed in each application group at 81 and 98 days after treatment, respectively. At six months, no signs of relapse were seen in remaining mice, indicating that this chemotherapy regimen was curative. Immunohistochemical (astrocytosis) and histological (inflammatory lesions) examinations of brain tissues showed that animals returned to normal from 2 months post-treatment. These results suggest that the megazol-suramin combination reversed the CNS pathology in this model. PMID- 9754670 TI - Meningitis caused by a serogroup W135 clone of the ET-37 complex of Neisseria meningitidis in West Africa. AB - Meningococci belonging to serogroup W135 caused several cases of meningococcal meningitis in The Gambia in 1995 and were isolated during a serogroup A epidemic in Mali in 1994. The eight isolates tested belonged to the same clone of the ET 37 complex and differed in several bands from the pulsed-field gel electrophoresis restriction pattern of serogroup C meningococci of the ET-37 complex isolated in Mali. Three of 6 patients infected in The Gambia died, indicating that this W135 clone is virulent. Vaccines that protect only against infections with meningococci belonging to serogroups A and C are usually used to control outbreaks in Africa, although vaccines containing the W135 polysaccharide are available. The findings of this study indicate that outbreaks of meningococcal meningitis in Africa can be associated with serogroup W135 infections and that serogrouping is essential before vaccination campaigns are started. PMID- 9754671 TI - Pooling sera to reduce the cost of HIV surveillance: a feasibility study in a rural Kenyan district. AB - Seroprevalence studies are crucial in HIV control programs but too expensive at district level. We evaluated the applicability of pooling sera and how it can reduce cost and affect accuracy at district level. 740 samples collected from antenatal clinic attendants for a sentinel survey in a rural Kenyan district were screened individually and in pools of 10. The seroprevalence when measured individually was 7.30%, while the calculated seroprevalence from pooled testing was 7.49%. Pooling was practicable and reduced costs by 62% for a marginal loss of accuracy. It is a useful tool in increasing the affordability of surveillance at district level. A pool size of 8 would have resulted in optimal cost reduction at minimal loss of accuracy. PMID- 9754673 TI - Seroprevalence of viral hepatitis in Tanzanian adults. AB - In a cross-sectional study in Dar es Salaam, Tanzania, we determined the seroprevalence of markers for hepatitis A, B, C and E viruses and examined associated risk markers. Among 403 healthy adults, the seroprevalence of antibodies to hepatitis A virus was 99.0% (95% confidence interval: 97.5-99.7). Prior exposure to hepatitis C and E viruses was rare (hepatitis C: 0.7% (0.2 2.1); hepatitis E: 0.2% (< 0.1-1.4)). The prevalence of all markers of hepatitis B was 70.7% (66.0-75.1). Hepatitis B surface antigen was identified in 6.0% (3.9 8.7) of subjects. Independent predictors of hepatitis B infection identified by logistic regression included older age, male gender, Muslim religion and type of abode. Given the high prevalence of hepatitis B and the low prevalence of hepatitis C, the majority of chronic viral hepatitis is likely to be associated with hepatitis B. Control efforts should focus primarily on hepatitis B. PMID- 9754672 TI - Measles vaccination before nine months. AB - OBJECTIVE: To measure the protective effect of measles vaccine administered before 9 months of age and compare overall mortality of children vaccinated at 6 8 months and at 9-11 months. METHOD: Non-concurrent cohort study involving all 13 134 children born between 16 January 1986 and 31st December 1991 in Kaniyambadi block near Vellore who had not left the area by six months of age. Main outcome measures were risk of disease and death among the under-five-year-olds according to age at measles immunization. RESULTS: Unimmunized children had a higher risk of developing measles compared to the immunized (P < 0.05). There was no significant difference in risk of measles among those vaccinated prior to and after nine months of age. Unvaccinated children were at significantly higher risk of death than vaccinated children (P < 0.001). There was no difference in risk of death between infants vaccinated between 6 and 8 months and those vaccinated between 9 and 11 months. There was no difference in the risk of death between boys and girls vaccinated between 6 and 8 months with standard-titre Edmonston Zagreb vaccine. CONCLUSION: Administration of standard-titre Edmonston-Zagreb measles vaccine at 6-8 months is an effective and safe preventive measure for measles, especially where the age-specific attack rate for children < 9 months is high. PMID- 9754674 TI - Influence of immunoadjuvants and a promiscous T-cell determinant on the immunogenicity of RESA peptide antigen of P. falciparum. AB - Synthetic peptide antigens representing the repeat sequences of malarial antigens showed poor immunogenicity and protection in clinical trials. In the present study, RESA, an asexual blood stage antigen, containing (EENVEHDA)2 and (DDEHVEEPTVA)2 sequences were chemically linked to a promiscous T-cell determinant (CS.T3) of the circumsporozoite protein of P. falciparum. The synthetic constructs either alone or coentrapped with immunoadjuvants (nor muramyl dipeptide/lauroyl tetrapeptide) were administered in liposomes to mice of varying genetic background and the immunogenicity of different formulations were compared under identical experimental conditions. The RESA peptide formulations containing the T-cell determinant and the adjuvants generated high titre and affinity antibodies in all the strains, as compared to peptide(s) alone. The booster immunization induced a strong anamnestic response in each group. Though the major IgG isotype is of IgG1 and IgG2b interestingly, formulations containing CS.T3 have a higher proportion of cytophilic IgG2b isotype. There was a significant fall in the levels of IgG2b isotype while IgG1 levels were maintained same in the third bleed (day 60, without booster immunization). The mixed peptide group preparation containing the adjuvant is found to be a better immunogen than that of respective peptides itself. The in vitro merozoite reinvasion inhibition assay showed 76-96% inhibition with the formulations containing RESA peptide(s) CS.T3 and the adjuvant, while with peptides alone the inhibition was 50-56%. This study highlights the importance of an alternative approach for developing peptide based immunogen against malaria. PMID- 9754675 TI - Combined treatments with Ninjin-youei-to (Ren-shen-yang-rong-tang) plus a suboptimal dose of prednisolone on autoimmune nephritis in MRL/lpr mice. AB - MRL/lpr mice suffer from a systemic lupus erythematosus-like autoimmune disease. We studied the effects of oral treatments with Ninjin-youei-to (NYT, Ren-shen yang-rong-tang, 1000 mg/kg/day), a suboptimal dose (2 mg/kg/day) of prednisolone(PSL) and their combination on nephritis in MRL/lpr mice. Treatments with NYT or PSL alone inhibited the development of proteinuria and prolonged survival. The combined treatment reduced the incidence of proteinuria and prolonged survival. In histological analysis, NYT treatments decreased the degree of mesangial proliferative glomerulonephritis and infiltration of mononuclear cells in the kidneys. PSL treatment was effective in reducing periglomerular nephritis and vasculitis in addition to such effects as NYT and NYT plus PSL treatment was more effective than PSL alone. The active form of TGF-beta was reduced in NYT and PSL-treated mouse serum, and the combined treatments further suppressed it. However, the treatment with NYT alone did not induce a decrease in the latent form of TGF-beta. The effect of NYT can be assumed to be different from an immunosuppressive effect of PSL. Therefore, the combined treatment with NYT and PSL can be expected to be more useful for the therapy of autoimmune disease such as nephritis, compared with NYT or PSL alone treatments. PMID- 9754676 TI - Inhibition of immediate type allergic reactions by the aqueous extract of Kum Hwang-San. AB - We studied the effect of aqueous extract of Kum-Hwang-San (KHS) on mast cell mediated immediate type allergic reactions. KHS (1-100 microg/site) inhibited concentration-dependently mast cell-dependent ear swelling response induced by compound 48/80 (200 microg/site) in mice by both topical and intradermal application. KHS (0.1-100 microg/site) inhibited concentration-dependently passive cutaneous anaphylaxis induced by anti-dinitrophenyl (DNP) IgE in rats by both topical and intradermal application. KHS also inhibited concentration dependently the histamine release from the rat peritoneal mast cells (RPMC) by compound 48/80 and anti-DNP IgE. Moreover, KHS had a significant inhibitory effect on anti-DNP IgE-induced tumor necrosis factor-alpha (TNF-alpha) secretion from RPMC. These results indicate that KHS inhibits immediate type allergic reactions by inhibition of histamine release and TNF-alpha secretion from mast cells in vivo and in vitro. PMID- 9754677 TI - Pharmacological effects of SA96 (bucillamine) and its metabolites as immunomodulating drugs--the disulfide structure of SA-96 metabolites plays a critical role in the pharmacological action of the drug. AB - SA96 (generic name, bucillamine) is a disease-modifying anti-rheumatoid arthritis (RA) drug with immunological effects. This compounds has two sulfhydryl groups in its molecule, and the differences and similarities between this drug and D penicillamine, which is also a sulfhydryl group-containing anti-rheumatic drug, have frequently been discussed. To clarify the pharmacological differences between these two drugs, we examined the concentrations of the compounds and its metabolites in serum and synovial fluid, paying special attention to the metabolites of SA96 produced in vivo. SA96 was metabolized in a very short time to SA981 which is a disulfide compound formed by intramolecular binding of two sulfhydryl groups, and transferred to synovial fluid. In addition SA981 had significant suppressive effects on IL-6 and IL-8 production by synovial cells in vitro. These results demonstrate that SA96, which has two sulfhydryl groups, exhibits anti-rheumatic effects via a pharmacological action clearly different from that of D-penicillamine. PMID- 9754678 TI - Tamoxifen exerts testosterone-dependent and independent effects on thymic involution. AB - Circulating testosterone concentrations and seminal vesicles weights, as well as thymus and spleen weights and histology were assessed in male Wistar rats from the infantile to post-pubertal period. The widely used anti-estrogenic agent tamoxifen was then administered in adult intact and castrated male rats and its long-term effects on thymic involution and splenic growth were examined. The results showed that: (1) age-related involution of the male thymus from the juvenile period through puberty to post-puberty depends on the rising testosterone levels and represents mainly a decrease of thymic lymphoid-cell elements; (2) tamoxifen administration reverses thymic involution in intact adult male rats and this effect is related to a dose-dependent, tamoxifen-induced castration and decrease of testosterone levels; (3) the changes of circulating testosterone levels, either resulting from maturity, or induced by tamoxifen or by castration, have a minimal effect on splenic growth and weight; and (4) in contrast to intact animals, administration of tamoxifen at pharmacological doses to adult castrated rats results in thymic regression. Underscoring the critical role of testosterone on thymic involution, these findings show that tamoxifen is able to reverse ageing changes in the thymus by suppressing testosterone production, while conversely, exerts thymolytic effects in the absence of androgens. PMID- 9754679 TI - Real time analysis of lung sounds. AB - This paper describes a real time system for the analysis of pulmonary sounds. The system performs various types of time-domain and spectrographic analysis. It is able to display time-domain waveforms obtained from microphones detecting lung sounds, their power spectra and a real-time linear prediction model instantaneously for the immediate identification of interesting features. Details of the system are presented with examples of clinical research carried out using spectrographic analysis. PMID- 9754680 TI - A new versatile PC-based lung sound analyzer with automatic crackle analysis (HeLSA); repeatability of spectral parameters and sound amplitude in healthy subjects. AB - A versatile PC-based lung sound analyzer has been developed for short-term recording and analysis of respiratory sounds in research and clinical applications. The system consists of two sound sensors, a flow sensor, a filtering signal amplifier and a PC with a data acquisition card and software for measurement and analysis of the sounds. The analyses include phonopneumography, time expanded waveform analysis, spectral analysis with time averaged Fast Fourier Transform, frequency analysis in time domain (sonogram), and automatic detection and waveform analysis of crackles. Short-term repeatability of spectral parameters of tracheal and lung sounds was studied in 10 healthy subjects. The coefficients of variation (CoV) of the averaged quartile frequencies (F25, F50 and F75) of lung sounds during flow-controlled tidal breathing were 3.7, 4.0 and 8.9% in expiration and 2.7, 3.5 and 4.5% in inspiration, respectively. CoVs of the averaged F25, F50 and F75 of expiratory tracheal sounds were 6.9, 3.0 and 2.4%, and those of inspiratory tracheal sounds 6.3, 2.6 and 3.3%, respectively. Examples of lung sound analysis of samples containing adventitious sounds such as crackles and wheezes are presented. The results indicate that the median frequency has the best repeatability of quartile frequencies of breath sounds and they suggest that the variations of those parameters are low enough for diagnostic purposes. The results also suggest that the analyzer can be a useful new tool for pulmonary research in the fields of physiological and clinical short term studies of respiratory sounds. PMID- 9754681 TI - Inverse filtering applied to upper airway sounds. AB - Inverse filtering is a digital signal processing technique which may be applied to speech-like sounds to remove resonances introduced by upper airway cavities to leave a residual signal which is, in principle, spectrally flat and strongly related to the excitation source. The filter parameters, normally computed by a form of linear prediction analysis, are indicative of the frequencies and bandwidths of the resonances. This paper briefly outlines the principle of inverse filtering and describes two applications in the study of upper airway sounds for diagnostic purposes. The first application is concerned with the non invasive measurement of variations in upper airway dimensions which occur with changes in posture. Results show that differences in the resonance frequencies caused by changes in posture can be measured, these being of the order of about 10% in normals. The measurement of such changes is known to be useful in the assessment of patients with sleep apnoea. The second application concerns the evaluation of vocal tract abnormalities resulting from infection in the larynx. Parameters derived from the residual are believed to be indicative of the existence and severity of a hoarse voice. Results have been obtained which support this theory. PMID- 9754682 TI - A new method for automatic wheeze detection. AB - A new automatic wheeze detection method which is based on image processing techniques applied to the sonagram was developed here. In the calculation of the sonagram, autoregressive and FFT spectrum estimation methods were compared. The method was validated in four wheezing asthmatic patients by a pulmonary physician. Nine out of ten wheezes longer than 250 ms were detected. Very short wheezes were not detected. The false positive amount of wheezing in control subjects was only about 1%. The method extracts also information about the frequency, duration, flow and volume associated with the wheezes. PMID- 9754683 TI - Wheezing Lung Sounds Analysis with adaptive local trigonometric transform. AB - Wheezes are abnormal sounds which are known to be relevant to Chronic Obstructive Pulmonary Diseases (COPD). The analysis of such signals is especially useful in patient monitoring or pharmacology. Respiratory sounds are dependent on the flow and the volume. Furthermore, they can be the result of a complex mixture of events. The analysis of lung sounds can be greatly improved with time-frequency techniques because these methods highlight the evolution of the spectra of events. In this paper, we present the application of the Adaptive Local Trigonometric Decomposition (ALTD) to lung sound analysis. This analysis provides an optimal representation of the signal in the time-frequency domain with a lattice which is adapted in time. In our work, the parameterization of the ALTD is studied for the detection of wheezing phenomena. PMID- 9754684 TI - Time-scale segmentation of respiratory sounds. AB - Respiratory sounds are composed of various events: normal and so-called adventitious sounds. These phenomena present a wide range of characteristics which make difficult their analysis with a single technique. Adapted time frequency and time-scale techniques allow to fit best, under constraints, the accuracy of analysis of a time segmentation and, by the way, make feasible the study of complex signals. We present here new approaches based only on the wavelet packet decomposition to segment respiratory sounds. PMID- 9754685 TI - Classification of respiratory sounds based on wavelet packet decomposition and learning vector quantization. AB - In this paper, a wavelet packet-based method is used for detection of abnormal respiratory sounds. The sound signal is divided into segments, and a feature vector for classification is formed using the results of the search for the best wavelet packet decomposition. The segments are classified as containing crackles, wheezes or normal lung sounds, using Learning Vector Quantization. The method is tested using a small set of real patient data which was also analysed by an expert observer. The preliminary results are promising, although not yet good enough for clinical use. PMID- 9754686 TI - A new method to detect crackles in respiratory sounds. AB - In this paper, an automatic method to detect and analyze crackles in digitised respiratory sounds is presented. The method is based on two steps: (1) a threshold (T) value is applied to the first derivative absolute value (FDAV) of lung sound to locate the "zone of interest" and (2) in this zone a crackle is detected if certain conditions are verified. The first derivative (FD) is evaluated by means of a derivative-smoothing filter, preserving areas under the spectral lines of the signal (moment zero), its mean position in time (first moment) and its spectral line width (second moment). The conditions to verify step 2 concern the following: the number and height of the peaks of FDAV and their distances from the starting point of the crackle, within a temporal window TW. This method shows good performances as an automatic detector (sensitivity 84% and specificity 89%), and is specifically designed to find the starting point of the crackle. PMID- 9754688 TI - Placing crackles on the flow-volume plane: a study of the relationship between the time position, the flow and the volume. AB - The aim of this study was to analyse inspiratory crackles in patients with Pulmonary Fibrosis (PF) and Bronchiectasis (BE). One case of Chronic Obstructive Pulmonary Disease (COPD) has also been included. The relationships between the time of occurrence of crackles (T) in the breath cycle and the corresponding flow at the mouth (F) and volume (V) have been investigated. The linear correlations between the flow, volume and time have been investigated by Pearson's R-test and the same variables have been analysed by Principal Component Analysis (PCA) in order to verify the effective dimension of these data. The results show a strong correlation between the time of occurrence and the volume in all the examined cases. PCA shows that in all cases F and V account for more than 90% of variation. These results suggest that placing crackles on the flow-volume plane does not cause loss of information. PMID- 9754687 TI - Distribution of crackles on the flow-volume plane in different pulmonary diseases. AB - A new method to represent and evaluate crackles on the flow-volume plane is described. Characteristic crackle patterns were found in patients with pneumonia, bronchiectasis, chronic obstructive pulmonary disease, heart failure and cryptogenic fibrosing alveolitis. In addition to visual assessment, simple statistical parameters were used to describe the observed pathological phenomena. PMID- 9754689 TI - Familial associations of subsyndromes of psychosis in affected sibling pairs with schizophrenia and schizoaffective disorder. AB - Attempts to describe the clinical heterogeneity of schizophrenia have consisted of categorical subtyping and a dimensional approach using factor analysis. The latter has yielded three dimensions or subsyndromes: positive, negative and disorganisation. The aim of this study is to explore to what degree these subsyndromes are correlated within 114 sibling pairs (185 individuals) with DSM III-R schizophrenia or schizo-affective disorder who were assessed for the lifetime presence or absence of the positive, negative, affective and disorganisation subsyndromes. Ratings were based on the core symptoms of each subsyndrome using a modified Krawieka scale. First rank symptoms were also included in the analysis. Coincidence was assessed by application of the binomial theorem to the frequency of occurrence of subsyndromes in this set of siblings. The disorganisation subsyndrome was shared above chance expectation (chi2=9.15, P < 0.01 for all sibling pairs). The significant results for the disorganisation subsyndrome suggest that it may be a suitable phenotypic marker for genetic linkage studies. PMID- 9754690 TI - Genotype-phenotype relationship in female carriers of the premutation and full mutation of FMR-1. AB - The present French-German cooperative study focuses on the genotype-phenotype relationship of mutations of the FMR-1 gene and psychiatric conditions in mothers with a full mutation in the FMR-1 gene of fra-X children (n=13), mothers with a premutation in the FMR-1 gene of fra-X children (n=61), as well as premutated siblings of these mothers without affected children (n=17) and two non-mutated control groups: (1) siblings of these mothers with normal CGG repeat (n=18); and (2) mothers of non-fra-X autistic children (n=42). Mothers with a full mutation in the FMR-1 gene and mothers with a premutation in the FMR-1 gene did not differ in the frequency of any axis I disorder; however, both groups were diagnosed with social phobia more often than the control group of mothers of autistic children. Moreover, mothers with a premutation in the FMR-1 gene of fra-X children and their siblings with the premutation (without affected offspring) revealed a similar frequency of social phobia. Furthermore avoidant personality disorder was more common in groups of carriers of the full premutation than in siblings without mutation or than the control group of mothers with autistic children. On the basis of our data, we therefore suggest that social avoidance (expressed as social phobia or avoidant personality disorder) has been underestimated in previous studies of carriers with the FMR-1 full mutation or premutation. Comorbidity of axis I and axis II psychiatric diagnoses was mainly restricted to the group of carriers of the full mutation and carriers of the premutation of FMR 1. Correlations between size of CGG repeat and IQ as well as CGG and age of onset of axis I diagnosis were non-significant. IQ of subjects had no impact on presence or absence of axis I and/or axis II diagnoses. PMID- 9754691 TI - Dopamine receptor D4 gene is associated with delusional symptomatology in mood disorders. AB - Disturbances of the dopaminergic neurotransmitter system have been implicated in the pathogenesis of depressive symptoms. Many studies have, however, failed to detect any association between genetic markers for the dopamine system and mood disorders. A possible reason for this may lie in the definition of phenotype, which is traditionally based on psychiatric diagnosis. In this study, we investigated the possibility that functional variants of the dopamine D4 receptor (DRD4) gene might be associated with depressive symptomatology in a sample of mood disorder subjects. Seventy-nine inpatients affected by bipolar (n=37) and major depressive (n=42) disorder (DSM-IV) were assessed at admission by the Hamilton Depression Rating Scale and were typed for DRD4 variants at the third exon using polymerase chain reaction (PCR) techniques. DRD4 was associated with delusional symptoms (F=5.56; d.f.=2,145; P=0.005), with DRD4*7 exhibiting higher scores when compared to DRD4*4 (P=0.006) variants. Polarity of mood disorder did not influence results significantly. The findings are in accordance with our previous report of an association of the DRD4 gene with delusional symptomatology of major psychoses. DRD4*7 should, therefore, be considered a liability factor for delusional symptoms in mood disorders. PMID- 9754692 TI - Self-esteem in remitted patients with mood disorders is not associated with the dopamine receptor D4 and the serotonin transporter genes. AB - Disturbances of the dopaminergic and serotoninergic neurotransmitter systems have been implicated in the pathogenesis of depressive symptoms. Associations have been reported between markers of the two neurotransmitter systems and the presence of illness or severity of depressive episodes, but no attention has been focused on the periods of remission. The present report focuses on a possible association of self-esteem in remitted mood disorder patients with the functional polymorphism located in the upstream regulatory region of the serotonin transporter gene (5-HTTLPR) and the dopamine receptor D4 (DRD4). Inpatients (N=162) affected by bipolar (n=103) and unipolar (n=59) disorder (DSM III-R) were assessed by the Self-Esteem Scale (SES, Rosenberg, 1965) and were typed for DRD4 and 5-HTTLPR (n=58 subjects) variants at the third exon using polymerase chain reaction (PCR) techniques. Neither DRD4 nor 5-HTTLPR variants were associated with SES scores, and consideration of possible stratification effects such as sex and psychiatric diagnosis did not reveal any association either. The serotonin transporter and dopamine receptor D4 genes do not, therefore, influence self esteem in remitted mood disorder subjects. PMID- 9754693 TI - No association between anxiety disorders and catechol-O-methyltransferase polymorphism. AB - Several studies have shown that the morbidity risk for anxiety disorders is increased among the relatives of patients with obsessive-compulsive disorder (OCD). Recently, it was reported that a polymorphism of the catechol-O methyltransferase (COMT) gene is significantly associated with OCD. The purpose of this study was to determine the association, if any, between the COMT polymorphism and anxiety disorders. We undertook an association study of the COMT polymorphism in 108 patients who met DSM-IV criteria for anxiety disorders and 135 healthy controls. All subjects were unrelated Japanese. The subdiagnostic groups did not differ significantly from the control group in either the genotypic or allelic frequencies. There were no statistically significant differences between the genotype and males, females, or a family history. The mean age of onset did not significantly differ among the genotypes. Our results suggest this functional COMT polymorphism does not make an important contribution to anxiety disorders in the Japanese population. PMID- 9754694 TI - An association of NAG levels and a mutation of the CCK gene in panic disorder patients. AB - Levels of the enzyme N-acetyl-beta-glucosaminidase (NAG) and a mutation of cholecystokinin (CCK) gene appear to be independently associated with panic disorder. We explored whether there was an association of NAG levels and a CCK mutation identified in a group of panic disorder patients. NAG was measured in 12 panic disorder patients who had a mutation of the CCK gene and 17 who did not. Urine for NAG was collected at baseline and after 3 and 6 weeks of treatment. NAG levels were lower at all three times in the patients that did not have the CCK mutation. The difference between the two groups was significant at week 6 (P < 0.02), and showed a trend toward a difference at baseline (P < 0.15). PMID- 9754695 TI - Autonomic instability during relaxation in panic disorder. AB - The ability to relax was assessed in 14 patients with panic disorder (PD) and 15 non-anxious control subjects for 10 min. Before and after relaxation, subjects performed a standardized activating task of talking continuously for 4 min. The fractional decline in reported anxiety, tension, and alertness between the first talking period and the relaxation minimum did not differ between groups, although absolute levels of anxiety and tension were higher for PD patients. The fractional decline in skin conductance between the first talking period and the last minute of relaxation was less for PD patients than control subjects, while their increase in skin temperature was greater. Skin conductance showed a linear decline over the logarithm of relaxation time, the slope of which was less steep for PD patients. Goodness of fit of skin conductance over log time was also significantly poorer for PD patients. Heart rate levels or slopes did not differ between groups. Autonomic differences between PD and control subjects were largely due to six patients who reported having panic attacks during the test and higher pretest anxiety levels. In conclusion, indicators of relaxation were inconsistent. Skin conductance suggested autonomic instability during quiet sitting in patients who panic or who are prone to panic. PMID- 9754696 TI - Working memory in childhood-onset schizophrenia and attention deficit/hyperactivity disorder. AB - We investigated verbal and spatial working memory in participants with childhood onset schizophrenia (N=13), attention-deficit/hyperactivity disorder (ADHD; N=31) and age-matched normal children (N=27). The ages of the participants ranged from 9 to 20 years, with an average age of approx. 14 in all groups. Diagnoses were based on structured interviews (Kiddie-Schedule for Affective Disorders and Schizophrenia) with the children and their parents and made using DSM-III-R criteria. Verbal working memory was assessed by the highest number of digits recalled in forward and backward order on the Digit Span subtest of the Wechsler Intelligence Scale. Results showed that normal children recalled more digits than schizophrenic and ADHD children, who did not differ. Spatial working memory was assessed with the Dot Test of Visuospatial Working Memory: The children were presented with a dot on a page for 5 s and asked to mark its location on a blank page immediately after presentation or 30 s later. A distracter task was used during the delay to prevent verbal rehearsal. The average distance between the target dot and the child's mark in the 30-s condition was shorter for normal than for schizophrenic and ADHD children, who did not differ. Thus, both schizophrenic and ADHD children showed deficits in verbal and spatial working memory. These results suggest that in both disorders, the capacity of the sensory buffers may be diminished, and/or the availability and allocation of resources to the central executive may be limited. PMID- 9754697 TI - Plasma neuropeptide-Y levels, monoamine metabolism, electrolyte excretion and drinking behavior in children with attention-deficit hyperactivity disorder. AB - Against a background of (a) increased drinking behavior in children with attention-deficit hyperactivity disorder (ADHD); (b) the parallel between some behaviors associated with ADHD and hypertension; (c) the use of the spontaneously hypertensive rat as a model for ADHD; and (d) similarities in the changes of neuropeptide Y (NPY) and catecholamine in studies of hypertension and drinking, NPY, catecholamines and electrolyte balance were compared in the plasma and urine of healthy children and those with ADHD. Drinking was monitored during 3 h of neuropsychological tests over 2 days in 14 ADHD and nine healthy children. Patients drank four times as much water and showed twice the levels of NPY found in controls. In controls there were positive and in patients there were negative relationships for NPY with drinking and restless behavior. Patients' plasma levels of norepinephrine (NE) and epinephrine were slightly elevated, but urinary levels of NE and the serotonin metabolite were markedly increased. Urinary excretion rates for sodium (not potassium), phosphate and especially calcium were decreased in patients even after covarying for less urine production in the ADHD group. NPY levels were inversely related to calcium excretion and drinking was inversely related to circulating sodium. Increases of drinking and circulating NPY in ADHD children and decreased electrolyte excretion may reflect a common disturbance in metabolic homeostasis. PMID- 9754698 TI - Anhedonia and relapse in alcoholism. AB - The aim of this study was to determine the role of anhedonia among other psychopathological dimensions in the relapse of alcoholics 6 months after withdrawal. Psychometric assessments included: the Social and Physical Anhedonia Scales, the Sensation Seeking Scale, the Pleasure-Displeasure Scale (including Fawcett-Clark's Pleasure Scale), the Depressive Mood Scale, the Thymasthenic Syndrome Rating Scale and the Comprehensive Psychopathological Rating Scale. Forty-four alcoholics participated in the study. The baseline values recorded during the second week of treatment showed that the more anhedonic the alcoholics were, the less they sought sensations. Type 2 alcoholics (Cloninger's classification) scored higher on the Thrill and Adventure Seeking Subscale. Relapsed alcoholics had higher baseline values on the Thrill and Adventure Seeking Subscale. This was in agreement with the step-wise discriminant analysis which showed that this subscale was the main variable that differentiated abstinent alcoholics from those who relapsed. Our results indicate that anhedonia does not predict relapse. PMID- 9754699 TI - Sample size and power calculations for comparing two independent proportions in a 'negative' trial. AB - Statistical methods for evaluating the adequacy of sample size and power cater mainly to the testing for treatment difference in a 'positive' trial. In biomedical research, the trial can sometimes be postulated as 'negative' to demonstrate that the different treatment groups are statistically equivalent. Herein we describe a computer program to determine the adequacy of sample size and power for comparing two independent proportions in a 'negative' trial. The program is written in MicroSoft QuickBasic Version 4.5, and its executable file is suitable for use as a stand-alone program on a microcomputer. PMID- 9754700 TI - A PET study of D2 dopamine receptor density at different phases of the menstrual cycle. AB - Behavioral and biochemical studies in animals indicate that central dopaminergic neurotransmission may be modulated by sex steroids. This may be the mechanism underlying the suggested association between estrogen and schizophrenia. The aim was to examine if different levels of sex steroids during the menstrual cycle are associated with variations in D2 dopamine receptor density as measured with positron emission tomography (PET) and [11C]raclopride. Five healthy women were examined, one during two subsequent follicular phases and four during two different phases of their menstrual cycle. In none of the women did the difference in putamen to cerebellum (P/C) ratios (-11 to 10%) exceed the difference in P/C ratios previously reported in test-retest analyses in men (-11 to 9%). The findings do not support the conclusion that there is a menstrual cycle-dependent variation in D2 receptor density detectable with single PET examinations and [11C]raclopride. Furthermore, a stable P/C ratio throughout the menstrual cycle indicated a stable D2 receptor occupancy in schizophrenic women treated with antipsychotic drugs. Repeated PET examinations of schizophrenic women known to deteriorate during particular phases of their menstrual cycle may further contribute to our understanding of the association between schizophrenia and sex steroids. PMID- 9754701 TI - Hemispheric control of motor function: a whole brain echo planar fMRI study. AB - The aim of this study was to explore whether recruitment of the ipsilateral motor cortex during non-dominant motor movement reflects left hemispheric control of motor function or simply the greater complexity or unfamiliarity of the motor task. BOLD fMRI was performed in normal right-handers during two motor tasks: (1) sequential finger movements (SM task) with the right or left hand; and (2) random finger movements (RM task) with the right hand. In all subjects, activation was predominantly in the contralateral motor areas (primary sensorimotor, lateral premotor, parietal and supplementary motor regions) and ipsilateral cerebellum. While the ipsilateral motor areas were also activated, single subject analysis revealed these areas to be more extensive and to be seen in more subjects during the non-dominant hand SM task and dominant hand RM task than during the more familiar dominant hand SM task. Similarly, group analysis also revealed ipsilateral activation in the primary sensorimotor and lateral premotor areas, but only during the non-dominant SM task and the dominant hand RM task. Non dominant hand movements, perhaps because they are less 'automatic', appear to require more cortical activity similar to complex tasks with the dominant hand, and result in greater recruitment of ipsilateral cortical motor areas and striatum. The study also illustrates how potentially meaningful subtleties seen on individual maps may be obscured with group averaging approaches. PMID- 9754702 TI - Reduced striatal dopaminergic innervation shown by IPT and SPECT in patients under neuroleptic treatment: need for levodopa therapy? AB - We investigated the availability of dopamine reuptake sites in the striata of two patients with productive symptoms and neuroleptic therapy as well as progressive parkinsonism using the new dopamine transporter ligand [123I]N-(3-iodopropen-2 yl)-2beta-carbo-methoxy-3beta- (4-chlorophenyl)tropane (IPT) and single photon emission computed tomography (SPECT). Normal specific binding in the caudate nucleus was 8.6 +/- 1.2 and in the putamen 6.5 +/- 1.3 (mean +/- S.D.; n = 8; mean age, 56.7 years; range 41-67 years). Patient 1 (age 43) was admitted to our clinic at age 38 because of left-sided parkinsonism. At age 40, she developed paranoid psychosis without change after cessation of L-DOPA and lisuride treatment for 3 months. She was diagnosed as a schizophrenic, paranoid subtype (DSM-III-R). IPT-SPECT showed a loss of dopaminergic nerve terminals (right caudate/putamen, 5.16/2.0; left caudate/putamen, 5.92/2.66). Patient 2 (age, 61 years) had a history of paranoid psychosis for approx. 30 years. He experienced progressive right-sided parkinsonism since age 57 when treated with clozapine. IPT-SPECT showed a marked reduction of striatal dopamine transporter binding (right caudate/putamen, 5.06/1.65; left caudate/putamen, 3.8/1.12). Our findings indicate that patients may suffer contemporaneously from Parkinson's disease and schizophrenia. In these patients, the proof of a nigrostriatal dopaminergic deficit justifies treatment with neuroleptics and dopaminergic drugs. Imaging of dopamine transporters with SPECT and IPT or a related compound represents an attractive alternative to the more complex measurements of fluorodopa uptake with positron emission tomography (PET). PMID- 9754703 TI - Differences in regional brain metabolic responses between single and repeated doses of methylphenidate. AB - Studies investigating the acute effects of drugs of abuse on human brain metabolism have measured single doses whereas these drugs are mostly taken repeatedly. Here we compared the brain metabolic response to intravenous methylphenidate when given after a single dose to that when given after two sequential doses. Methylphenidate-induced changes in metabolism differed; whereas single doses tended to decrease metabolism, repeated doses tended to increase it, and these differences were significant in frontal, parietal and occipital cortices and hippocampus. This indicates that methylphenidate's metabolic effects vary with acute previous exposure and highlights the importance of studying drugs after single and repeated administration. PMID- 9754704 TI - Cytokine and interferon research in Israel. AB - From its inception, the field of interferons and cytokines has occupied an important position in Israeli biological science. With the Second Joint Meeting of the International Society for Interferon and Cytokine Research and the International Cytokine Society taking place in Jerusalem in 1998, it is timely to review here current Israeli research on the biology, gene regulation, receptors, signal transduction, mode of action and clinical aspects of interferons and cytokines. PMID- 9754705 TI - Interleukin-18: perspectives on the newest interleukin. AB - Just over two years ago the newest member of the interleukin family of cytokines, IL-18, was molecularly cloned. IL-18 was originally identified as a result of its ability to induce interferon gamma production, however with the advent of its cloning and the production of recombinant protein a number of other biological actions have since been identified. Recently the receptor for IL-18 was also characterised. Due to the structural and biological properties shared between IL 18 and IL-1 and their respective receptors, questions relating to IL-18 activities are being answered at a rapid pace. This article addresses the biology of IL-18 in both disease and non-disease states. PMID- 9754706 TI - IFN-gamma expression in macrophages and its possible biological significance. AB - IFN-gamma is a pleiotropic cytokine endowed with potent immunomodulatory effects whose expression was long considered to be restricted to T and NK cells. Only recently, it became evident that IFN-gamma production can also occur in other cell types, including monocyte/macrophages. However, the biological relevance of macrophage IFN-gamma is still unclear. The purpose of this article is to provide an overview of the collected evidence demonstrating IFN-gamma expression in macrophages and to discuss the possible biological significance of this cytokine production in the early phase of host response to infectious agents. PMID- 9754707 TI - Pleiotropic functions of neurotrophins in development. AB - Neurotrophins are soluble growth factors known mainly for their roles in regulating the development of the mammalian nervous system. Two types of receptors mediate the actions of these polypeptides: the Trk family of tyrosine kinase receptors and the so-called p75 low-affinity NGF receptor. Neurotrophins and their receptors are highly expressed in the nervous system. Gene targeting approaches in the mouse have uncovered some of their functions in promoting survival and developmental maturation of certain types of neurons of the peripheral and central nervous system, confirming their critical role in neural development. Furthermore, the phenotypes observed in these mutants have demonstrated the specificity of the interactions between neurotrophins and their receptors. These families of genes are also widely expressed in a variety of non neuronal systems throughout development, including the cardiovascular, endocrine, reproductive and immune systems. Our knowledge of neurotrophin functions in non neuronal tissues is still fragmented and mostly indirect. Nevertheless, there is increasing evidence that neurotrophins may have broader physiological effects besides regulating neuronal survival and differentiation. Analysis of mice lacking neurotrophins or neurotrophin receptors promises to provide avenues for elucidating these functions. PMID- 9754708 TI - The role of helper T cell subsets in autoimmune diseases. AB - CD4 helper T cells can be divided into Th1 and Th2 subsets based upon the cytokines they produce. Th1 and Th2 cells have been found to be mutually antagonistic, leading to either Th1- or Th2-dominated responses upon immunization. In recent years, several authors have suggested that in chronic inflammatory autoimmune diseases such as diabetes, multiple sclerosis and rheumatoid arthritis, Th1 cells are pathogenic and Th2 cells are protective. Therefore, a successful deviation from a Th1-dominated to a Th2-dominated response could have clinical benefits for individuals suffering from these diseases. Unfortunately, data accumulated over recent years have not supported this approach, in particular regarding the protective role of Th2 cells. In this review we discuss these data and conclude that, at least using currently available tools, immune deviation from Th1 to Th2-dominated responses is ineffective unless started at very early (subclinical) stages of the disease. In addition, we examine some recent data suggesting that, under some circumstances, Th2 cells can be pathogenic. PMID- 9754710 TI - Chemokines in disease models and pathogenesis. AB - Investigators from a wide variety of disciplines met at the Second National Managed Health Care Congress Meeting on chemokines held in Washington, D.C. on December 14-15, 1997, to discuss the role of chemokines in the pathogenesis of disease states, as well as a number of biological issues. Presentations on the effects of chemokines in animal models were interspersed with talks on fundamental chemokine structure-function relationships, signal transduction, the role of chemokine in cell trafficking, inflammation, immunity and hematopoietic development. Although it was impossible to consider the score of chemokine receptors and the 50 or more chemokines cloned to date, most of the more well established and some of the newer chemokines were discussed. We will first summarize the preconference symposium on the role of chemokines in neurobiology and then review the various issues addressed by the other speakers to provide a more integrated rather than sequential summary of the proceedings. PMID- 9754709 TI - Keratinocyte growth factor: a unique player in epithelial repair processes. AB - Keratinocyte growth factor (KGF) is a member of the rapidly growing fibroblast growth factor (FGF) family of mitogens. Whereas most FGFs influence proliferation and/or differentiation of various cell types, KGF seems to act specifically on epithelial cells. It has been demonstrated that KGF stimulates proliferation and migration of these cells, but it also affects differentiation processes. Finally, recent studies have demonstrated a protective function of this growth factor in vitro and in vivo. Due to these properties, KGF could play an important role in repair processes. Indeed a series of studies have provided insight into the expression and function of KGF in inflammation and repair of various tissues and organs, and a therapeutic potential of this growth factor has been demonstrated. PMID- 9754711 TI - Recent advances in cytokines, cytokine receptors and signal transduction. AB - The Fifth Annual Conference of the International Cytokine Society was held on November 9-13, 1997 at Lake Tahoe, Nevada. This meeting kept up with the tradition of exciting talks and posters, presenting significant advances in our understanding of the cytokine world. As we advance our knowledge, a complex network of interacting cellular communication pathways is revealed. Targeted disruption of genes coding for cytokines, their receptors and their cytoplasmic signaling molecules, became the standard method for a proper assessment of the role of a given cytokine in vivo. Yet, fundamental questions remain unresolved. For instance, it is not yet known how signal specificity is maintained when different cytokine receptors use the same cytoplasmic signaling pathways. The following summary is not comprehensive, rather, it is a collection of representative communications. Many more top quality studies were presented at the meeting and we apologize for not being able to review all of them here. PMID- 9754712 TI - The G-protein betagamma complex. AB - The vast majority of signalling pathways in mammalian cells are mediated by heterotrimeric (alpha betagamma) G proteins. Reviewed here is regulation of signal transduction by the betagamma complex at different protein interfaces: subunit-subunit, receptor-G protein and G protein-effector. The role of diverse beta and gamma subunit types in achieving specificity in signalling and potentially unidentified functions for these subunits also are discussed. PMID- 9754713 TI - Crowded little caves: structure and function of caveolae. AB - Caveolae are small vesicular invaginations of the cell membrane. It is within this organelle that cells perform transcytosis, potocytosis and signal transduction. These "little caves" are composed of a mixture of lipids and proteins unlike those found in the plasma membrane proper. The chief structural proteins of caveolae are caveolins. To date, three caveolins (Cav-1, -2 and -3) with unique tissue distributions have been identified. Caveolins form a scaffold onto which many signalling molecules can assemble, to generate pre-assembled signalling complexes. In addition to concentrating these signal transducers within a distinct region of the plasma membrane, caveolin binding may functionally regulate the activation state of caveolae-associated signalling molecules. PMID- 9754714 TI - Alternations of diacylglycerol kinase in streptozotocin-induced diabetic rats. AB - Dysfunction of organs has been reported in diabetic rats, suggesting an association with changes in intracellular signal transduction pathways including phosphatidylinositol (PI) turnover. Diacylglycerol (DG) kinase catalyses the phosphorylation of DG, which is considered to play a major physiological role in the metabolism of the intracellular messenger DG. However, no relation between DG kinase activity and any disease in mammalian tissue has been reported to date. In the present study, we investigated whether the changes in DG kinase activity are related to diabetes. Basal resting level of DG kinase activity changed in tissue isolated from diabetic rats. Decreases in resting activity detected in aorta and kidney and agonist-induced responses differed between these tissues. Submaximal increases in basal activity also were detected in vas deferens and hepatocytes. These changes in DG kinase activity resemble the functional changes associated with complications of diabetes, suggesting that changes in PI turnover followed by DG kinase activity are a key element in the complications. It is the first study about the changes in DG kinase activity in mammalian disease. PMID- 9754715 TI - Tumour necrosis factor stimulates stress-activated protein kinases and the inhibition of DNA synthesis in cultures of bovine aortic endothelial cells. AB - In this study, we examined the ability of tumour necrosis factor-alpha (TNF) to stimulate the mitogen-activated protein (MAP) kinase homologues p42/44 MAP kinase, c-Jun NH2-terminal kinase (JNK) and p38 MAP kinase and its effect upon DNA synthesis in primary cultures of bovine aortic endothelial cells (BAECs). TNF strongly stimulated p38 MAP kinase and JNK activity in both a time- and concentration-dependent manner. By contrast, TNF was a very poor activator of p42/44 MAP kinase relative to the known activator of p42/44 MAP kinase in endothelial cells, adenosine triphosphate (ATP). TNF-stimulated activation of p38 MAP kinase, and MAPKAP kinase-2, a known downstream target of p38 MAP kinase, was strongly inhibited by pre-incubation with the p38 MAP kinase inhibitor SB203580, whereas the minor activation of p42/44 MAP kinase was abolished by pre-incubation of the cell with the novel MAP kinase kinase 1 inhibitor PD098059. Addition of TNF resulted in a 50-60% decrease in DNA synthesis in BAECs. Pre-incubation with PD098059 or co-incubation with ATP failed to modify the inhibitory effect of TNF upon DNA synthesis. SB203580 reduced basal DNA synthesis by approximately 50%; however, if failed to modify the inhibition mediated by TNF. These results indicate that TNF strongly activates both p38 MAP kinase, JNK and, to a minor extent, p42/p44 MAP kinase. It is likely that only one of these kinases, JNK, plays a role in the regulation of DNA synthesis in these cells. PMID- 9754716 TI - Analysis of choline and phosphorylcholine content in human neutrophils stimulated by f-Met-Leu-Phe and phorbol myristate acetate: contribution of phospholipase D and C. AB - ABSTRACT. We analysed changes in choline (CHO) and phosphorylcholine (PCHO) content of stimulated human polymorphonuclear leukocytes (PMNs) by a chemiluminescence assay to further examine the relative contributions of phospholipase D (PLD) and PLC to phosphatidylcholine (PC) breakdown. PLD activation was also analysed by measuring tritiated phosphatidic acid (PA) and diglycerides (GDs) in PMNs labelled with tritiated alkyl-lyso PC. Stimulation of PMNs with formyl-methionyl-leucyl-phenylalanine fMLP; 0.1 microM induced a weak elevation of mass choline (+25% of basal level) that was strongly potentiated in PMNs primed with cytochalasin B (+350% relative to the control value of 657+/-53 pmol/10(7) cells). CHO production was rapid and transient, peaking within 1 min, and ran parallel to that of tritiated PA. Thereafter, the amount of tritiated PA declined strongly (40% of maximum by 3 min), whereas the elevated choline content induced by fMLP plateaued for at least 5 min. Phorbol myristate acetate (PMA) sustained the formation of CHO for as long as 20 min, which correlated with that of [3H]PA in a time- and concentration-dependent manner. PCHO content of resting PMN leukocytes (1560 +/- 56 pmol/10(7) cells) was not modified after stimulation of PMNs with fMLP or PMA for at least 10 min, which argues against breakdown of phosphatidylcholine by PLC. For longer treatment (10-20 min), fMLP stimulated a significant enhancement of PCHO level, which occurred concomitantly with a decrease in CHO level, suggesting that choline kinase rather than PLC may be activated. Unlike fMLP, PMA stimulated a fall in PCHO between 10 and 15 min after PMN stimulation, pointing to different regulatory mechanisms of PCHO level. These data indicate that DG formation from PC in PMNs is mediated by PLD but not by PLC and show that chemiluminescence measurement of choline is a reliable index of PLD activation. PMID- 9754717 TI - Critical role of conserved histidine pairs HNXXH and HDXXH in recombinant human phosphodiesterase 4A. AB - Cyclic AMP-Phosphodiesterases (cAMP-PDEs) catalyse the hydrolysis cAMP to AMP and thus serve to modulate the ligand-->adenylate cyclase-->cAMP-->PKA signal transduction pathway. PDEs exist as a multigene family of enzymes that bear significant sequence homology in the catalytic domains. The sequence alignment of these domains has revealed the presence of two histidine motifs: motif I, HNXXH, and motif II, HDXXH. These amino acid sequences are canonical motifs, which act as ligands for divalent metal cations required for catalytic activity. In this paper, we report human monocyte PDE4A to be a zinc-binding protein. Substitution by site-directed mutagenesis of either histidine in motif I by serine, which is not a ligand for metals, results in complete loss of catalytic activity and loss of sensitivity to divalent metal cation activation. However, similar mutations in motif II gave proteins that retained both approximately 50% of initial activity and the ability to respond differentially to Mg2+, Mn2+ and Co2+. Moreover the motif II mutants exhibited both functional group requirements and retained their pKa values. When the inactive mutants were affinity-labelled with 8-BDB-TcAMP and probed with antibody against cAMP or antibody against PDE4A, Western blots were unaltered. These results show that the conserved histidines in motif I are an absolute requirement for catalytic activity, whereas motif II histidines are required only to achieve maximum activity. PMID- 9754718 TI - Extracellular H+ stimulates the expression of c-fos/c-jun mRNA through Ca2+/calmodulin in PC12 cells. AB - Evidence has accumulated that an increase in extracellular protons stimulates the transmembrane mechanism to induce various intracellular responses, such as the expression of c-fos and c-jun. In the present study, we aimed to obtain evidence that an increase in extracellular protons induces expression of c-fos/c-jun mRNA in PC12 pheochromocytoma cells of rats. We found that the c-fos/c-jun mRNA expression increased when extracellular pH was decreased gradually from 7.40 to 7.20 and that there was a significant correlation between extracellular pH values and the expression of c-fos/c-jun mRNA. To determine whether the Ca2+/calmodulin system subserves the H+-induced expression of c-fos/c-jun, Ca2+/calmodulin inhibitor trifluoperazine was added to PC12 cells. We found that trifluoperazine inhibited the expression of the H+-induced c-fos/c-jun mRNA by 30-35%. In contrast, trifluoperazine did not inhibit the expression of phorbol-induced c fos/c-jun mRNA. These results indicate that an increase in extracellular protons induces the expression of c-fos/c-jun mRNA, and this expression is mediated partly by the Ca2+/calmodulin system. PMID- 9754719 TI - Diadenosine polyphosphate-mediated activation of phospholipase D in isolated rat liver cells. AB - Diadenosine polyphosphates (ApnAs) can, through interaction with appropriate purinoceptors, affect a range of cellular activities. Ap4A, the most prominent naturally occurring diadenosine polyphosphate, stimulates alterations in intracellular calcium homeostasis and subsequent activation of glycogen breakdown in isolated liver cells. Here we show that Ap4A, and other naturally occurring diadenosine polyphosphates, also stimulates phospholipase D (PLD) activity in isolated rat liver cells. The characteristics of Ap4A-mediated activation of PLD are similar to those for the activation of PLD by extracellular ATP. These results are discussed in the context of the relation between diadenosine polyphosphate- and adenine mononucleotide-mediated cellular signalling processes. PMID- 9754720 TI - Transforming growth factor beta 1 potently activates CPP32-like proteases in human hepatoma cells. AB - Induction of apoptosis in Hep3B hepatoma cells by transforming growth factor beta 1 (TGF-beta 1) was accompanied by the activation of interleukin-1-beta-converting enzyme-like proteases, which have recently been renamed as caspases. The caspase inhibitor ZVAD-FMK, which has a broader specificity for caspase family proteases, blocked TGF-beta 1-induced apoptosis in a concentration-dependent manner. The caspases in this apoptotic process were further characterized by using a more specific caspase inhibitor, DEVD-FMK, for CPP32-like (caspase-3-like) proteases. Our results demonstrated that CPP32-like proteases were activated during apoptosis triggered by TGF-beta 1. Enhancement of CPP32-like activity was clearly detected in TGF-beta 1-treated Hep3B cells. Furthermore, cleavage of poly(ADP ribose) polymerase, an in vivo substrate for CPP32, in these cells was confirmed by immunoblotting. Thus, we suggest that CPP32-like proteases participate in apoptotic cell death induced by TGF-beta 1. PMID- 9754722 TI - Non-steroidal anti-inflammatory drugs and the chemoprevention of gastrointestinal cancers. PMID- 9754721 TI - Correlation between Bcl-X expression and B-cell hybridoma apoptosis induced by activin A. AB - In this study, we examined the role of bcl-XL and bcl-XS in apoptotic cell death of HS-72 cells induced by activin A. Immunoblot analysis revealed that a band of Bcl-XL was detected in HS-72 cells cultured with or without activin A. Although untreated HS-72 cells did not express Bcl-XS, the expression of Bcl-XS was significantly increased when cultured with activin A. We also investigated the expression of Bcl-XS and Bcl-XL in HS-72 cells cultured with activin A in the presence of protein kinase C inhibitor 1-(5-isoquinotinesulfonyl)-2 methylpiperazine dihydrochloride (H7), which suppressed apoptosis in HS-72 cells induced by activin A. Exposure to H7 apparently increased the level of Bcl-XL in HS-72 cells cultured with or without activin A. In contrast, no detectable band of Bcl-XS was found in HS-72 cells cultured with activin A and H7. These findings indicate that Bcl-XL upregulation and Bcl-XS downregulation induced by H7 might correlate with the suppression of activin A-induced apoptosis in B-lineage cells. PMID- 9754723 TI - Role of acid and salivary epidermal growth factor in gastric mucosal adaptation to naproxen in man. AB - BACKGROUND: Non-steroidal anti-inflammatory drugs (NSAIDs) frequently damage the gastrointestinal tract, but with continued administration this usually resolves by a process of adaptation. There is evidence that the acute injury can be reduced by acid suppression, and animal models have shown that salivary epidermal growth factor (EGF) is an important factor in gastric mucosal adaptation. We therefore wanted to assess the effect of acid suppression and salivary EGF output during naproxen-induced acute gastric injury and subsequent adaptation. METHODS: Healthy subjects were given a 14-day course of naproxen with different regimens of ranitidine and placebo. Before and on three occasions during treatment subjects provided a salivary sample for EGF and underwent gastroscopy to assess gastric damage. RESULTS: Similar gastric damage occurred after 24 h in all groups and resolved in most subjects. Base-line salivary EGF output was similar in all groups but increased in the placebo/ranitidine group on day 3 and in the ranitidine group on day 9. CONCLUSIONS: Acid suppression with ranitidine did not prevent acute gastric injury. Adaptation may be associated with an increase in salivary EGF output. PMID- 9754724 TI - Animal reservoirs in the transmission of Helicobacter heilmannii. Results of a questionnaire-based study. AB - BACKGROUND: Infection with either Helicobacter heilmannii or H. pylori causes gastritis in humans. Whereas the route of transmission of H. pylori appears to be human-human, it has been suggested that H. heilmannii is transmitted by animals. METHODS: Two hundred and two patients with histologically confirmed H. heilmannii gastritis and 600 randomly selected patients with H. pylori gastritis were sent a questionnaire, in which they were asked whether they had had or still have regular contact with dogs, cats, rodents, horses, birds, cattle, pigs, or wild animals. RESULTS: The questionnaires of 177 patients with H. heilmannii and 485 with H. pylori infection were evaluated. In comparison with H. pylori, H. heilmanii infection was strongly associated with contact with dogs, cats, cattle, or pigs. Logistic regression analysis showed that contact with pigs, cats, and dogs leads to a significant risk of H. heilmannii infection (odds, 4.990, 1.710. and 1.462). CONCLUSION: Pigs, cats, and dogs appear to be reservoirs in the transmission of H. heilmannii. PMID- 9754725 TI - Management of dyspeptic patients in primary care. Value of the unaided clinical diagnosis and of dyspepsia subgrouping. AB - BACKGROUND: Most dyspeptic patients in primary care are managed without confirmatory investigations. In this study the reliability of the unaided clinical diagnosis and the diagnostic value of dyspepsia subgrouping are evaluated in unselected dyspeptic patients in primary care. METHODS: Six hundred and twelve unselected dyspeptic patients were referred for interview and endoscopy. General practitioners stated a provisional diagnosis and a proposed management strategy. Before endoscopy, patients were classified on the basis of predominant symptoms as reflux-, ulcer-, or dysmotility-like or as unclassifiable RESULTS: The sensitivity and the positive predictive value of the diagnosis of ulcer were 0.58 and 0.29, respectively, and those for esophagitis 0.30 and 0.43. The predictive value of a clinical diagnosis of functional dyspepsia was high, but, considering the high prevalence of the condition, the chance-corrected validity was at the same level as for the other diagnoses (0.18-0.22). Classification of patients by predominant symptoms increased the a priori probability of ulcer and esophagitis in the respective subgroups. However, more than one-third of the patients with ulcer or esophagitis were classified in inappropriate subgroups. CONCLUSIONS: It is difficult to select an appropriate management strategy for dyspeptic patients on the basis of symptoms and history alone. Dyspepsia subgroups are of limited help in the decision process because of the low predictive value of the endoscopic diagnosis. PMID- 9754726 TI - A study of intestinal dysfunction induced by restraint stress in rats. AB - BACKGROUND: The pathogenesis of intestinal dysfunction induced by stress has not been established. We tried to clarify possible causal mechanisms of irritable bowel syndrome. METHODS: An experimental model of intestinal dysfunction was designed using loading restraint stress in rats. A cannula was inserted into the origin of the duodenum or colon, with the other end leading to the skin. To provide intestinal content, a semi-solid colored marker was used for monitoring intestinal transit. After 1 week the marker was injected into the intestine through the cannula under unanesthetized wakefulness. RESULTS: Under restraint stress, transit in the small intestine was suppressed, but actual suppression took place only in the upper half, where the contents normally moved fast. Transit time in the colon was reduced by restraint stress. This reduction was attributed to the disappearance of the stagnant region, which was present under normal conditions. CONCLUSIONS: These results suggested that restraint stress affects the function of the pacemaker site of the intestine. PMID- 9754727 TI - Nonsteroidal anti-inflammatory drug-associated upper gastrointestinal lesions in rheumatoid arthritis patients. Relationships to gastric histology, Helicobacter pylori infection, and other risk factors for peptic ulcer. AB - BACKGROUND: Nonsteroidal anti-inflammatory drugs (NSAIDs) are risk factors for peptic ulcer in rheumatoid arthritis (RA) patients, but the contribution of reactive gastritis, concomitant Helicobacter pylori infection, or RA activity to NSAID ulcer pathogenesis is unknown. METHODS: Ninety-six RA patients taking NSAIDs and dyspeptic sex- and age-matched control patients without NSAID use or an RA diagnosis were enrolled in the study. RESULTS: Gastric ulcer (GU) was detected in 29 (30%) RA patients and 3 control patients (P < 0.001). Sixteen RA patients and no control patient had an H. pylori-negative GU. The GUs of the RA patients were mainly located in the prepyloric region (28%) and antrum (62%). Nine of the 29 RA patients (31%) with GU had more than 1 ulcer. Erosive gastropathy was detected in 34 (71% H. pylori-negative) RA patients and in 13 (62% H. pylori-negative) control subjects (P < 0.001). Chronic gastritis was observed in 65 RA patients (48% H. pylori-negative) and in 58 control subjects (43% H. pylori-negative) (NS). whereas reactive gastritis was found in only 2 RA patients and in none of the controls. Corticosteroid use was the only independent risk factor for GU: odds ratio was 6.8 (95% confidence interval, 1.3-36.0). The prevalences of duodenal ulcer or esophagitis were not increased in RA patients. CONCLUSIONS: RA patients using NSAIDs continuously are at a greatly increased risk of developing both H. pylori-negative and -positive GUs, and corticosteroid use is an independent risk factor for ulcer development. Most RA patients have chronic gastritis, whereas reactive gastritis is rarely associated with continuous NSAID use in RA patients. PMID- 9754728 TI - Serum IgA antibodies from patients with coeliac disease react strongly with human brain blood-vessel structures. AB - BACKGROUND: Our objective was to determine the possible presence of IgA antibodies directed against human central nervous system (CNS) structures in sera from coeliac disease (CD) patients. METHODS: Serum samples were collected from 4 patients with active CD on a gluten-containing diet, 11 biopsy-proven CD patients on a gluten-free diet (GFD), and 52 non-coeliac gastrointestinal controls. In all patients IgA antigliadin antibody (AGA) titres were determined with enzyme-linked immunoassay (ELISA), and IgA antiendomysium antibodies (EMA) with indirect immunofluorescence on human umbilical cord. Cryostat sections of human brain occipital cortex were incubated with the patients' sera and subsequently labelled with anti-human IgA fluorescein conjugate. RESULTS: All sera from patients with active CD on a gluten-containing diet yielded positive results in both the IgG AGA and EMA test and in indirect immunofluorescence on brain tissue, disclosing a strong fluorescence over blood-vessels structures. All sera from CD patients on a GFD and from non-coeliac gastrointestinal controls gave a negative result on both the EMA test and the immunofluorescence reaction on human brain. CONCLUSIONS: Sera from patients with active CD contain IgA antibodies that react with human brain vessel structures, giving intense fluorescence. These antibodies are not present in sera from coeliac patients on a GFD or non-coeliac controls. This finding might be involved in the abnormal nervous system manifestations frequently described in association with coeliac disease. PMID- 9754729 TI - Timing of postprandial dyspeptic symptoms and transpyloric passage of gastric contents. AB - BACKGROUND: Patients with functional dyspepsia often experience early satiety and discomfort after a meal. The role of early gastric emptying in symptom generation is not known. Our aim was to relate timing of symptoms and early postprandial emptying in functional dyspepsia. METHODS: Twelve patients with functional dyspepsia were investigated during 3 min of fasting, during 3 min of ingesting 500 ml of a meat soup, and during the first 10 min postprandially by means of duplex sonography. RESULTS: Gastric emptying commenced on average 52 sec after the start of ingestion. Transpyloric movements of gastric contents unrelated to peristalsis (that is, alternating transpyloric emptying and reflux within a common chamber created by the terminal antrum, the pylorus, and the duodenal bulb) appeared before peristaltic-related emptying, which commenced after on average 116 sec. In all patients epigastric, meal-related discomfort was experienced after the commencement of transpyloric emptying, on average after 143 sec. A negative correlation was found between intensity of fullness and duration of presymptomatic transpyloric movements of gastric contents (that is, the duodenal 'tasting' period). CONCLUSIONS: The early occurrence of meal-related symptoms suggests that gastric distension is the main factor in symptom generation. However, the onset of symptoms after the commencement of gastric emptying suggests that intestinal tasting receptors are involved in symptom generation. The inverse relationship between the duration of the tasting period and symptom intensity suggests that the time allowed for duodenal tasting might be too short in patients with FD. PMID- 9754730 TI - Glucagon-like peptide-2 inhibits centrally induced antral motility in pigs. AB - BACKGROUND: Glucagon-like peptide-2 is formed from proglucagon in the intestinal L-cells and is secreted postprandially in parallel with the insulinotropic hormone GLP-1 (glucagon-like peptide-1), which in addition acts to inhibit gastric motility (enterogastrone effect) by inhibiting central parasympathetic outflow. GLP-2 has no effect on the endocrine pancreas. We here tested the hypothesis that GLP-2 acts as an enterogastrone. METHODS: Fourteen anesthetized pigs with their splanchnic nerves cut were subjected to insulin hypoglycemia, and force transducers were sutured to the antrum to record motility. GLP-2 was infused intravenously in doses from 1 to 6 pmol/kg/min after the onset of antral motility in response to hypoglycemia. RESULTS: Insulin hypoglycemia invariably and greatly increased the frequency and amplitude of antral phasic contractions. Infusions of GLP-2 dose dependently (1-6 pmol/kg/min) inhibited antral motility. At 2 pmol/kg/min, resulting in plasma GLP-2 concentrations of 102.5+/-19 pmol/l (normal postprandial range, 30-82 pmol/l), the motility index was inhibited by 91%+/-14%. CONCLUSIONS: Both of the intestinal glucagon-like peptides may operate as hormonal transmitters of the ileal brake effect. PMID- 9754731 TI - Primary culture and transfection of epithelial cells of human small intestine. AB - BACKGROUND: So far, no techniques are available for primary culture and efficient transfection of human small-intestinal enterocytes, which would provide a valuable tool to investigate intestinal function. METHODS: Human small-intestinal biopsy specimens were treated with collagenase and dispase. Resulting crypt units were cultured for several days. Using the intestinal epithelial cell lines Caco-2 and HT-29, we established optimal conditions for transfection of a control plasmid, which were then applied to primary cultured cells. RESULTS: Cells growing out of crypt units formed monolayer-like sheets and proliferated for several days. Most of the cells could be stained with antibodies against epithelial markers. Among seven different transfection reagents tested, Lipofectamine was the most potent, with transfection efficiencies up to 25% for primary enterocytes. CONCLUSIONS: An easy technique was developed providing viable small-intestinal enterocytes that can be efficiently transfected. PMID- 9754732 TI - Adult patients receiving home parenteral nutrition in Denmark from 1991 to 1996: who will benefit from intestinal transplantation? AB - BACKGROUND: Small-bowel transplantation is an alternative to home parenteral nutrition (HPN) in patients with gut failure. Our aim was to report the indication, diagnosis, morbidity, mortality, and intestinal adaptation in the total cohort of Danish patients receiving HPN at any time during the 5 years between 1 January 1991 and 31 December 1995. The data were analysed against the option of transplantation. RESULTS: HPN was given to 129 patients; 59 (46%) had inflammatory bowel disease (15% died), 26 (20%) had cured cancers (42% died), and 44 (34%) had other diseases (dysmotility, surgical complications, infarcts, and so forth; 27% died). Of these, 60% were new in the HPN program, but only 19% received HPN all 5 years; 31 % had terminated HPN, 19% permanently, and 25% died. Only four deaths were HPN-related. In December 1995, 73 patients were receiving HPN in Denmark, for a prevalence of 13.9 per million, which is the highest in Europe but 10-fold lower than in the United States. CONCLUSIONS: Gut failure was the only indication for HPN in Denmark. Weight loss without gut failure, such as disseminated cancer and acquired immunodeficiency syndrome, was not an indication for HPN. Survival after small-bowel transplantation should be assessed against a sizeable mortality among candidates receiving HPN, and this depends on diagnosis and age. In an HPN population comparable with the Danish, a quarter is likely to die within a period of 5 years, a quarter will terminate HPN, and the others survive with HPN. Small-bowel transplantation can be a lifesaving procedure in the small fraction of foreseeable HPN-related deaths, mainly caused by liver failure. Transplantation will not improve survival in most adult HPN patients, and only an improved quality of life after transplantation justifies this procedure in most HPN patients. PMID- 9754733 TI - Impairment of phagocytic and T-cell-mediated antibacterial activity and plasma endotoxins in patients with untreated gastrointestinal cancer. AB - BACKGROUND: Cancer patients have multiple immune deficits, and mediators, such as prostaglandins, transforming growth factor-beta, and interleukin (IL)-10, may play a role in the pathogenesis of these immune dysfunctions. METHODS: Fifty-six patients with gastrointestinal cancer (11 gastric cancer, 7 papilla of Vater cancer, and 38 colorectal cancer) were enrolled for this study, before starting conventional treatments. Phagocytosis and killing exerted by polymorphonuclear cells and monocytes, peripheral blood mononuclear cell absolute numbers, T-cell mediated antibacterial activity, serum levels of IL-10 and interferon (IFN) gamma, and plasma bacterial endotoxin concentration were evaluated. RESULTS: Data show an impaired phagocytic and T-cell-mediated antibacterial activity in all cancer patients, whereas only in subjects with gastric cancer were IFN-gamma serum levels reduced. Circulating endotoxins were detected in 17 patients. CONCLUSIONS: In untreated gastrointestinal cancer patients the capacity of phagocytes and T-cells to clear pathogens is reduced. This dysfunction may increase the risk of becoming infected and may account for the presence of endotoxin in 30% of patients. PMID- 9754734 TI - Effects of branched-chain-enriched amino acid solution on insulin and glucagon secretion and blood glucose level in liver cirrhosis. AB - BACKGROUND: The aim of this study was to determine whether therapeutically used branched-chain amino acids (BCAAs) solution influences glucose metabolism in liver cirrhosis (LC). METHODS: BCAAs solution (200 ml) was infused in LC patients at different stages, and plasma concentrations of glucose and pancreatic hormones were determined. RESULTS: In patients with mild LC, BCAAs caused a significant increase in glucose level (maximal increment, 12.5+/-2.5 mg/dl) with a great increase in insulin (maximal increment, 39.5+/-8.3 microU/ml) and a small increase in glucagon secretion (maximal increment, 101.0+/-16.0 pg/ml). In patients with advanced LC, BCAAs caused a great increase in glucagon secretion (220.5+/-19.4 pg/ml) with only a slight increase in glucose levels (5.8+/-2.2 mg/dl). CONCLUSION: BCAAs solution causes hyperglycemia in mild LC due to insulin resistance, whereas it causes only a slight increase in severe LC due to hepatic glucagon resistance. Thus, there is a possibility that BCAAs solution may lead to hypoglycemia in advanced LC with hepatic glycogen depletion. PMID- 9754735 TI - Increased plasma levels of substance P and disturbed water excretion in patients with liver cirrhosis. AB - BACKGROUND: The pathogenesis of impaired water excretion in liver cirrhosis has not been fully elucidated. METHODS: We induced an intravenous water overload of 20 ml/kg body weight in 10 cirrhotics without ascites (CLC), 11 cirrhotics with ascites (DLC), and 10 normal subjects (N) and investigated the relationship of plasma levels of substance P (SP), norepinephrine (NE), and antidiuretic hormone (ADH) to impaired water excretion. RESULTS: Free water clearance (CH2O) was lower in DLC (mean, 2.7 ml/min) than in N (8.3 ml/min; P < 0.001) and CLC (6.9 ml/min; P < 0.001). In DLC the creatinine clearance (CCr), maximal urine flow rate/CCr, (CH2O + CNa)/CCr, and mean arterial pressure (MAP) were significantly lower than in N and CLC. There was a progressive increase in basal SP, from lowest in N to CLC, to highest in DLC. Basal NE increased in CLC and DLC. Basal ADH did not differ among N, CLC, and DLC. In cirrhotics CH2O was correlated positively with serum albumin and cholinesterase and negatively with the retention rate of indocyanine green at 15 min. Basal SP was negatively correlated with CH2O (r= 0.71: P < 0.001) and MAP (r= -0.56; P < 0.005). Basal NE was correlated positively with basal SP (r= 0.67, P < 0.01 ). CONCLUSIONS: Decreased CH2O is closely related to the severity of the liver disturbance. Decreased CCr and reduced delivery of filtrate to the ascending limb of the loop of Henle secondary to an increased sodium reabsorption in the proximal tubule may play an important role in the impairment of water excretion. The increase in SP, which has a potent vasodilatory action, and the associated enhanced activity of the sympathetic nervous system may be responsible for the mild or moderate impairment of water excretion in the absence of nonosmotic hypersecretion of ADH in cirrhotics with ascites. PMID- 9754736 TI - Small-intestinal bacterial overgrowth in patients with liver cirrhosis, diagnosed with glucose H2 or CH4 breath tests. AB - BACKGROUND: Small-intestinal bacterial overgrowth (SIBO) has been considered a predisposing factor of spontaneous bacterial peritonitis in cirrhotic patients by bacterial translocation or hematogenous spread during spontaneous bacteremia. We investigated 45 cirrhotic patients and 28 healthy subjects to assess the prevalence of SIBO and its relationship with the severity of liver dysfunction and the presence of ascites. METHODS: Bacterial overgrowth was measured by the glucose hydrogen and methane breath test. RESULTS: SIBO was documented in 16 (35.6%) of the 45 cirrhotic patients and in 1 (3.6%) of the 28 healthy controls. The prevalence of SIBO was significantly higher in patients with Child-Pugh class B or C (50%) than in those with class A (19%) and had no relationship with the presence or absence of ascites. CONCLUSIONS: We conclude that the prevalence of SIBO in cirrhotic patients is approximately 35.6% and that it is related to the severity of liver disease. There was no difference among various causes of cirrhosis, such as viral, alcoholic, or idiopathic. PMID- 9754737 TI - T-cell receptor Valpha gene expression of infiltrating T cells in pancreatic cancer. AB - BACKGROUND: T lymphocytes play a central role in the immune response to cancer, with specific T-cell reactivity provided by the T-cell receptor (TCR) alphabeta chain heterodimer. Whereas human pancreatic adenocarcinoma is characterized by a massive infiltration of T lymphocytes, to date no analysis of the TCR Valpha-gene expression of the tumor-infiltrating lymphocytes in pancreatic cancer has been performed. METHODS: Using reverse transcriptase polymerase chain reaction (RT PCR) followed by dot blot hybridization, we determined the TCR alpha-chain repertoire at the mRNA level in pancreatic carcinoma and compared our findings with the TCR Valpha repertoire in the normal pancreas and chronic pancreatitis. RESULTS: A heterogeneous lowly restricted TCR Valpha repertoire was observed in pancreatic carcinomas, different from the TCR Valpha repertoire in chronic pancreatitis. CONCLUSIONS: Pancreas-infiltrating T cells show a distinct TCR Valpha gene expression profile in normal pancreas, chronic pancreatitis, and pancreatic cancer. PMID- 9754738 TI - Long-term follow-up of patients with chronic pancreatitis in Italy. AB - BACKGROUND: We investigated the epidemiologic, clinical, and radiologic aspects of a mixed medicosurgical series of chronic pancreatitis patients observed at the University of Verona Centre for the Study of Pancreatic Diseases over the period 1971-1995. METHODS: Even though the pathogenesis of chronic pancreatitis has yet to be clarified and the classification is still debatable, the patients were subdivided in accordance with the Marseilles-Rome classification into those with alcoholic, obstructive, familial, and idiopathic forms of the disease. A total of 715 patients were analysed with a median follow-up period of 10 years (range, 1 25 years). RESULTS AND CONCLUSIONS: At the end of follow-up the dropout rate amounted to 7.1% (51 patients), and 176 patients (24.6%) had died. Data are reported about the type of chronic pancreatitis, sex, and age distribution at the onset of the disease, drinking and smoking habits before onset and during follow up, and incidence of calcifications, pain, diabetes, steatorrhoea, and pseudocysts. Surgical aspects, survival curves, and causes of death are also analysed, and the most frequent concomitant diseases in chronic pancreatitis sufferers are discussed. PMID- 9754739 TI - Failure of Sengstaken balloon tamponade for rebleeding after tissue adhesive injection in a fundic varix. AB - A 61-year-old man developed a huge fundic varix due to portal hypertension in alcoholic liver cirrhosis. After a third injection therapy session with tissue adhesive (Histoacryl) massive hemorrhage developed. Sengstaken (gastric) balloon tamponade failed. Autopsy showed a huge, solid varix with a large hole on its side, inaccessible with the Sengstaken balloon. PMID- 9754740 TI - Detection of Helicobacter pylori in faeces with a new enzyme immunoassay method: preliminary results. PMID- 9754741 TI - The beetle Anthrenus verbasci causing proctitis and perianal itching. PMID- 9754742 TI - Psychological effects of aesthetic dental treatment. AB - OBJECTIVE: To demonstrate that aesthetic restorative dental treatment, using the porcelain laminate veneer, has a positive effect on the self-esteem of a patient. METHODS: A study group of 17 patients, unhappy with their dental appearance, were assessed psychologically at the pre-operative, immediate post operative and 6 month review stages. Porcelain laminate veneer restorations were used to improve the dental aesthetics for the patients in the study group. A comparison group of 27 subjects, without a dental appearance problem, were also psychologically assessed at comparable intervals. The assessments included Cattell's 16PF Personality Inventory, a Body-Esteem index, a computer administered version of the Repertory Grid technique and semi-structured interviews. RESULTS: The results showed that there were no significant differences between the study and comparison groups on any first or second-order factor of the 16PF. However, highly significant changes in a positive direction (p < 0.005) were observed in the study group in responses to a Body-Esteem questionnaire at each stage in the procedure. Comparison group changes were not significant. Repertory Grid analyses showed a significant overall convergence in the study group compared with the comparison group. There were also significant positive shifts amongst the study group in the normalised ratings of "self' on several of the constructs. Interviews confirmed the Repertory Grid findings. CONCLUSIONS: The study shows that aesthetic restorative treatment has a positive effect on patients' self esteem. PMID- 9754743 TI - Marginal integrity and postoperative sensitivity in Class 2 resin composite restorations in vivo. AB - INTRODUCTION: Problems that may arise in resin composite Class 2 restorations include microleakage and postoperative sensitivity. However, limited in-vivo research is conducted to evaluate these processes. AIM: The aim of this study was to assess postoperative sensitivity, microleakage and the pooling of adhesives in relation to Class 2 box-type composite restorations placed in vivo using various adhesive systems and application techniques. MATERIALS AND METHODS: One hundred and forty-four Class 2 box restorations were placed in the mesial and distal surfaces of 72 premolar teeth in-vivo using one of three combinations of adhesive systems and three filling techniques. After 6 weeks of clinical service postoperative sensitivity was recorded. The teeth were then extracted, immersed in a dye solution and sectioned. Microleakage and pooling of the adhesive was recorded. Statistical analysis involved logistic regression and chi2 tests to identify differences between groups at p < 0.05. RESULTS: Of the 144 restorations, 65 showed minimal cervical leakage in enamel, 5 suffered leakage into dentin and 74 were free of microleakage. No statistically significant differences were found in cervical microleakage between the adhesive systems or between filling procedures. Occlusal microleakage in the enamel was present in 16 of the 160 restorations. Liner Bond 2 restorations leaked significantly more at the occlusal surface (p < 0.05). Pooling of the adhesive was significantly less when PhotoBond was used. No spontaneous postoperative sensitivity was reported. Twenty-eight restorations were sensitive to loading. Postoperative sensitivity was significantly less in patients with Liner Bond 2 restorations. CONCLUSIONS: The adhesive systems used in this study showed minimal leakage into dentin in vivo. Using Liner Bond 2, restorations exhibited more occlusal leakage but were significantly less sensitive to loading. PMID- 9754744 TI - Survival of three types of veneer restorations in a clinical trial: a 2.5-year interim evaluation. AB - OBJECTIVES: In this clinical trial, 180 veneer restorations (VRs) were evaluated. The purpose of the study was to collect survival data and to find possible relations between survival and (1) 'type of VR', (2) 'preparation design', (3) 'operator' and (4) the patient-related variables 'tooth-type' and 'vitality of the tooth'. METHODS: The restorations were provided by seven dentists in 1 12 patients on central and lateral maxillary incisors. Experimental variables were: 'type of VR' (either direct resin composite (DC), indirect resin composite (IC) or porcelain (P)), 'preparation design' (with and without incisal overlap) and 'operator'. Failures were recorded at two levels: absolute failure (need for new restoration) and relative failure (need for repair). Survival was defined at three levels: (1) survival of original restoration (Sr, endpoints: 'absolute' failures), (2) functional survival (Sf, endpoints: 'relative' failures) and (3) overall survival (SO, endpoints: both 'absolute-' and 'relative failures'). RESULTS: The variable 'type of VR' showed significant influence on Sf and So but not on Sr. Sf and So rates of P, IC and DC were, respectively: Sf-P, 94%; So-P, 94%; Sf-IC, 94%; So-IC, 90%; Sf-DC, 80%; So-DC, 74%. VRs on vital teeth showed a significantly better survival than VRs on non-vital teeth at all survival levels. CONCLUSIONS: Preparation of the incisal edge for incisal coverage is considered to be unnecessary to assure or improve the strength of VRs. Veneers on non-vital teeth showed higher risk to fail than veneers placed on vital teeth. Porcelain veneers showed the best overall survival. PMID- 9754745 TI - Dimensional changes during veneering procedures on discoloured teeth. AB - OBJECTIVES: The aim of the study was to determine the dimensional changes of teeth when veneering procedures are involved. METHODS: Fifteen discoloured central incisors were selected from a group of 180 veneer restorations (VRs) composed of three different types of materials (direct resin composite, indirect resin composite and porcelain). Impressions and dies were made before treatment, after preparation and after placement and finishing the restoration. Contact stylus profilometry and subsequent analysis of the 3-D surface images provided quantitative data of the differences between the various treatment phases. RESULTS: The preparation reduction was the most for indirect resin composite VRs and the least for direct resin composite VRs. All veneer restorations showed nearly the same thickness and dimensional change after treatment, except one, which resulted in an additional increase of volume of the tooth. CONCLUSION: From the results of this study it is concluded that the dimensions of a discoloured tooth treated with a veneer restoration unintentionally increased, resulting in overcontour. PMID- 9754746 TI - Adherence of Candida albicans to denture-base materials with different surface finishes. AB - OBJECTIVES: To assess the in vitro adherence of Candida albicans to heat-cured hard and soft denture-base materials with varying surface roughness, and to observe the effect of a mixed salivary pellicle on candidal adhesion to these surfaces. METHODS: In vitro adhesion assays on heat-cured acrylic resin (Trevalon), Molloplast B and Novus using the type strain of C. albicans (NCPF 3153A). Surfaces for the assays were prepared using clinically appropriate rotary instruments. Unstimulated, pooled and clarified whole saliva was used to assess its effect on adhesion. RESULTS: Significantly greater adhesion of C. albicans to rough rather than smooth surfaces was found (P < 0.001), as well as increased adhesion to the machined soft lining materials compared with acrylic. Pre-coating denture-base materials with saliva reduced candidal adhesion on all materials. CONCLUSIONS: Rough surfaces on denture-base materials promote the adhesion of C. albicans in vitro. However, saliva reduces adhesion of C. albicans and thus diminishes the effect of surface roughness and free surface energy differences between materials. PMID- 9754747 TI - Decoloration of 1% methylene blue solution in contact with dental filling materials. AB - OBJECTIVES: Leakage studies have been performed frequently, since a tight seal provided by various dental fillings has been considered clinically important. The dye penetration experiment using a methylene blue solution as a tracer is one of the most common methods applied in these types of studies. The stability of the colour of methylene blue in contact with six dental filling materials was observed. METHODS: Silicon rubber tubes and human tooth roots of 10 mm in length and 1.5 mm inner diameter were filled with amalgam, calcium hydroxide, Cavit, Fuji II, mineral trioxide aggregate, or zinc oxide eugenol, 10 tubes or roots for each material. Groups of five tubes or roots filled with the same material were immersed in 0.8 ml 1% methylene blue dye solution. The optical density of the methylene blue solution before immersion and after 24, 48 and 72 h of immersion was measured in a spectrophotometer at 596 nm. RESULTS: The methylene blue solution was found to be decoloured over time by all the test materials (P < 0.01) except for Fuji II, in both silicone tubes and roots. At 24 h, the optical density value of methylene blue decreased by 73% for the Ca(OH)2/silicone group and 84% for the mineral trioxide aggregate/silicone group. CONCLUSION: Methylene blue is decoloured by some dental filling materials, which may result in unreliable results for these materials in dye leakage studies. PMID- 9754748 TI - The uptake and release of fluoride by ion-leaching cements after exposure to toothpaste. AB - OBJECTIVES: The cariostatic action associated with the glass-ionomer cement (GIC) is usually attributed to its sustained release of fluoride. However the ability of the GIC to act as a fluoride reservoir, taking it up from an external source (e.g. toothpaste, mouthwash) and subsequently releasing it over time, may also be a contributory factor. This study investigated the reservoir effect of various recently introduced ion-releasing cements: two resin-modified glass-ionomer cements (Fuji II LC, Vitremer), a compomer (acid-modified composite resin) (Dyract), and a recently introduced conventional glass-ionomer (Fuji IX). METHODS: Specimens were exposed to a fluoridated toothpaste after 28 and/or 58 days. The release of fluoride into the storage water, both before and after exposure, was monitored using a differential electrode cell. RESULTS: There was no significant difference in the fluoride releases from Vitremer and Fuji II LC. These materials released significantly more fluoride than Fuji IX and Dyract. All the materials released more fluoride on the day after exposure to an external fluoride source compared with the day before exposure. Release rates returned to baseline within 3 days. Within the time periods of the study, only the uptake/re release of Fuji IX was adversely affected by late exposure. All the materials showed an enhanced uptake and release on repeated exposure to the external fluoride source. CONCLUSIONS: All the materials under test (Dyract, Fuji II LC, Vitremer and Fuji IX) released significant amounts of fluoride and reacted positively to exposure to an external fluoride source. PMID- 9754749 TI - Retention of posts with resin, glass ionomer and hybrid cements. AB - OBJECTIVES: To measure and compare the retention of serrated root canal posts cemented with glass ionomer, resin and resin-modified glass ionomer (hybrid) cements. METHODS: Fifty single-rooted human teeth were decoronated, treated endodontically and then embedded in resin blocks. Standard post-holes, 10 mm long, were prepared to receive 1.5 mm serrated stainless steel posts. Five equal sized groups of roots had posts cemented using either a glass ionomer cement, one of two resin cements or one of two resin-modified glass ionomer luting cements. The cements were prepared and used according to the manufacturers' instructions. The tensile force required to dislodge the cemented posts in a testing machine was recorded. Statistical analysis was performed using Student's t-test and Mann Whitney U-tests at the 99.9% confidence level. RESULTS: Statistical analysis revealed that posts cemented with resin A were significantly better retained (340.06 N+/-23.13 N) than those cemented with resin B (212.56 N+/-67.62 N), or either of the two resin-modified glass ionomer cements (53.90 N+/-28.42 N, 25.97 N+/-14.70 N), but not statistically better than posts cemented with the glass ionomer cement (286.16 N+/-38.71 N). The retention of posts cemented with either resin B or the glass ionomer cement was significantly better than with either hybrid cement. There was no significant difference in retention between the hybrid cements. CONCLUSION: The performance of the resin-modified glass ionomer cements was significantly below that of alternative cements in this study. Possible explanations for this finding are discussed. Dentists should be cautious in adopting this new cementing regime. PMID- 9754751 TI - Regional bond strengths of self-etching/self-priming adhesive systems. AB - The purpose of this study was to measure the regional tensile bond strengths (TBS) at various portions of human tooth enamel and dentin, and to observe the resin-dentin interfaces of two commercially available self-etching/self-priming adhesive systems by scanning electron microscopy (SEM). Twelve extracted human cuspid teeth were used to measure TBS and four additional teeth were used for scanning electron microscopy (SEM). Outer enamel and dentin were removed from the labial tooth surfaces to form a long cavity preparation into middle dentin extending from the mid-crown to the apex of the root within the same tooth. Either Clearfil Liner Bond 2 (LB 2, Kuraray) or Fluoro Bond (FB, Shofu) was bonded to the surfaces, and covered with resin-composite. The resin-bonded teeth were serially sliced at right angles to the long axis of the tooth, and the bonded surfaces were trimmed to give a bonded cross-sectional surface area of 1 mm2 for TBS tests. LB 2 and FB showed significantly higher TBS in coronal, cervical and middle root dentin than that in enamel and apical root dentin. SEM showed that the thickness of the hybrid layer of both systems was about 1.0 microm in coronal, cervical and middle root dentin, and less than 0.5 microm in apical root dentin. These results suggested that the self-etching/self-priming systems produce good adhesion in coronal, cervical and middle root dentin by creating thin hybrid and transitional layers, but bonding to enamel and apical root dentin should be improved. PMID- 9754750 TI - Metal chloride primers for bonding dentine with tri-n-butylborane-initiated luting agents. AB - OBJECTIVES: The bonding of resin to dentine is dependent largely on surface modifications. The purpose of the present study was to test the hypothesis that a role of the metal chlorides used as primers is to initiate polymerization of resin at the resin-dentine interface, thereby affecting bond strength. METHODS: Nine primers were evaluated, comprising aqueous 2-hydroxyethyl methacrylate (HEMA) solutions containing AICl3, CeCl3, CoCl2, CuCl2, FeCl3, NiCl2, MgCl2, SnCl2 or ZnCl2, respectively. One of the two luting agents (Super-Bond resin) consisted of methyl methacrylate (MMA), 4-methacryloyloxyethyl trimellitate anhydride (4-META) and tri-n-butylborane (TBB) initiator. The other luting agent (MMA-TBB resin) consisted of MMA and TBB without 4-META. Extracted bovine teeth were ground to expose the dentine, etched with an aqueous solution of 10% phosphoric acid, primed, and then bonded with stainless-steel rods; tensile bond strengths were determined after 1-day immersion in water. Data were analysed by ANOVA and Duncan's new multiple range test (P < 0.05). RESULTS: Four of the metal chlorides (CuCl2, FeCl3, MgCl2 and ZnCl2) enhanced the bond strengths of MMA-TBB resin to dentine. With Super-Bond resin, maximum bond strengths of 15.6 MPa and 19.0 MPa were recorded with primer containing 2.0 x 10(-5) mol g(-1) FeCl3 and 2.0 x 10(-6) mol g(-1) CuCl2, respectively. CONCLUSIONS: Bonding techniques, combining the use of either cupric primer or ferric primer with 10% phosphoric acid etchant may be suitable for application in seating resin-bonded prostheses with TBB-initiated luting materials. PMID- 9754752 TI - Tensile, shear and cleavage bond strengths of alginate adhesive. AB - OBJECTIVES: This study determined the effect of alginate adhesive on various bond strengths of alginate to stainless steel. METHODS: Three test assemblies were designed and machined in stainless steel for tension, shear and cleavage tests. Alginate adhesive (Fix) was applied thinly and dried for 5 min. Alginate (Blueprint) was then loaded and allowed to set for 5 min before testing. The force at failure was measured by an Instron machine with a cross-head speed of 50 mm min(-1). RESULTS: Breaking stresses of alginate without adhesive were found to be 65 kPa (tension), 31 kPa (shear) and 10 kPa (cleavage). The bond strengths of Blueprint with Fix were 100 kPa (tension), 42 kPa (shear) and 37 kPa (cleavage) giving improvements of 53%, 37% and 270% respectively (p < 0.05). CONCLUSIONS: Alginate adhesive increases the bond strength of alginate, particularly cleavage, to stainless steel. PMID- 9754753 TI - Survival rate of ceramic inlays. AB - OBJECTIVES: The aim of the present study was to evaluate the survival rate of ceramic inlays provided in a practice environment by one of the authors over the past decade. METHODS: 183 inlays were examined in 67 patients. The interval between placement and assessment was, on average, 4 years (s.d. 2.75 years) and varied from 4 months to 10 years. Kaplan-Meier survival-type curves were used to assess the survival rate. RESULTS: Five inlays failed: four due to endodontic reasons and one due to fracture. Four failures were in permanent molar teeth while the other was in a premolar tooth. A success rate of 97% at 10 years was estimated. CONCLUSIONS: The clinical durability of the resin-bonded ceramic inlays investigated was satisfactory. PMID- 9754754 TI - Therapeutic node dissections in malignant melanoma. AB - BACKGROUND: Therapeutic lymphadenectomies involve the dissection and removal of clinically enlarged, histologically positive nodes at the regional nodal basin, in the absence of detectable distant disease. METHODS: The literature dealing with therapeutic lymphadenectomies in malignant melanoma was reviewed. RESULTS: The rate of wound complications varies with the particular nodal basin. The 5 year survival varies from 19% to 38%, with an average of 26%. Survival is affected primarily by the number of histologically positive nodes and extracapsular spread, and secondarily by the extent of disease at the various levels of the nodal basin, fixation of the nodes, and, probably, the preceding disease-free interval. Prognostic parameters of the primary lesion, e.g., thickness, ulceration, and location, also may have an effect on survival. The rate of local recurrence at the nodal basin after lymphadenectomy has varied from 0.8% to 52%. Adjuvant therapy with interferon alfa-2b has improved the 5-year disease-free survival from 26% to 37%. CONCLUSIONS: Therapeutic node dissections in melanoma provide an appreciable 5-year survival rate, which is further augmented by adjuvant therapy. Many series report a significant rate of local recurrence at the nodal basin following therapeutic dissection. Complete lymphadenectomy reduces the rate of local failure with its attendant morbidity. PMID- 9754755 TI - The Society of Surgical Oncology and the Commission on Cancer: progress through synergism. AB - The 1998 Presidential Address highlights the history of The Society of Surgical Oncology and the Commission on Cancer of the American College of Surgeons, cites specific examples of progress through synergism, and discusses some of the many challenges facing surgical oncologists in the future. These include the necessity to synergize in clinical trials, to accelerate the diffusion of knowledge into the practicing community, and to redefine surgical oncology and its relation to general surgery and the American Board of Surgery. PMID- 9754756 TI - Results of immunoscintigraphy using a cocktail of radiolabeled monoclonal antibodies in the detection of colorectal cancer. AB - BACKGROUND: External immunoscintigraphy using a single monoclonal antibody has been employed successfully to localize primary, recurrent, and occult colorectal carcinoma. This prospective study investigated the accuracy and sensitivity of external immunoscintigraphy when the combination or "cocktail" of radiolabeled monoclonal antibodies, CYT-103 (an IgG1a) and CYT-372 (an IgG2b) directed against TAG-72 and CEA, respectively, is given to patients with known or suspected colorectal cancer. METHODS: Eleven patients enrolled in this open label phase I/II study underwent preoperative external immunoscintigraphy after intravenous cocktail administration of two indium 111-labeled monoclonal antibodies (MoAb), CYT103 and CYT372. Antibody dose ranged from 0.2 mg (five patients) to 1.0 mg (six patients), each antibody radiolabeled with 2.5 mCi of indium 111, delivering a total dose of 5 mCi per patient. Planar and SPECT images were performed 2 to 5 days postinjection. Suspected lesions were surgically resected within 2 weeks of injection. RESULTS: A total of 23 lesions (sites) were identified in the eleven patients, 19 of which were confirmed by pathology (hematoxylin and eosin [H&E]). Cocktail immunoscintigrams identified 16 of the 19 confirmed lesions. Computed tomography (CT) scan detected 9 of the 19 lesions. The sensitivities of cocktail immunoscintigraphy and CT scan for the detection of colorectal cancer were 84% and 64%, respectively. The positive predictive value for immunoscintigraphy was 94%. The antibody scans detected six occult, previously unsuspected lesions. Cocktail immunoscintigraphy changed the surgical management in four of the 11 (36%) patients. CONCLUSIONS: The combination of In 111 CYT-103 and CYT-372 improved the sensitivity of external immunoscintigraphy for the detection of colorectal cancer compared to that obtained with a single MoAb imaging. Cocktail antibody imaging may enhance the staging and management of patients with cancers of colon and rectum. PMID- 9754757 TI - Overexpression of CD44: a useful independent predictor of prognosis in patients with colorectal carcinomas. AB - BACKGROUND: The goal was to investigate the potential correlation between overexpression of CD44, high microvessel count (MVC), and p21ras with length of relapse-free and overall survival in patients with colorectal adenocarcinomas. METHODS: CD44, factor VIII-related antigen (FVIII-RA), and p21ras were localized immunohistochemically in patients with colorectal adenomatous polyps (n = 8) and adenocarcinomas (n = 98). The correlation between the expression of CD44, MVC in the areas with highest density, and p21ras with relapse-free and overall survival time was investigated. Data were analyzed statistically using univariate and multivariate systems. RESULTS: In patients with adenomatous polyps, the positivity of CD44, FVIII-RA, and p21ras was 75%, 62%, and 88%, respectively. In patients with colorectal carcinomas the positivity of CD44 was 55%, and for p21ras it was 52%. The median of FVIII-RA was 4 MVC (range, 0.0 to 32.33). MVC was greater than 4 in 53% of the patients with colorectal carcinomas. In univariate analysis, a significantly longer relapse-free time (CD44: P = .0004; FVIII-RA: P = .0006) and overall survival time (CD44: P = .0001; FVIII-RA: P = .001) were observed for patients with CD44-negative tumors and MVC below 4 as compared to those with CD44-positive tumors and MVC greater than 4. Similar observations were noted in patients with Dukes B and C disease and the rectum as the site of tumor. In multivariate analysis, only CD44 correlated significantly with both relapse-free (P = .0003) and overall survival (P = .00001). CONCLUSION: Univariate analysis showed CD44 and MVC to be independent predictors of prognosis in colorectal carcinomas. Multivariate analysis showed that CD44 positivity was the most important indicator of an unfavorable prognosis for relapse-free and overall survival in patients with colorectal cancer. Thus, it can be deduced that whether CD44 is positive or negative in patients with colorectal cancer may have prognostic importance and in the future may be used as a factor in the pathologic evaluation of tumor specimens. This hypothesis needs to be tested prospectively in a larger number of patients. PMID- 9754758 TI - Surgery versus surgery and postoperative radiotherapy in squamous cell carcinoma of the buccal mucosa: a comparative study. AB - BACKGROUND: The efficacy of postoperative radiotherapy for squamous cell carcinoma of the buccal mucosa was evaluated. METHODS: One hundred seventy-six patients treated between 1989 and 1993 were analyzed. One hundred fifteen patients were treated with surgery alone (Group 1), and 61 patients were treated with a combination of surgery and postoperative radiotherapy (Group 2). RESULTS: Actuarial 3-year locoregional control in Groups 1 and 2 was 11% and 48% for patients with stage III + IV cancer (P = .001) and 71% and 75% for patients with stage I + II cancer (P = .74), respectively. On multivariate analysis for locoregional failure, surgical margin, bone invasion, high grade, and node involvement were significant factors in Group 1, whereas in Group 2 only tumor thickness was a significant factor. For local failure, margin, bone invasion, and stage in Group 1 and tumor thickness in Group 2 appeared as significant factors. For nodal failure, clinical nodal (cl N0 vs. N+) stage and grade in Group 1 and pathologic nodal stage (pN0 + 1 vs. pN2) in Group 2 were observed as significant factors. On subset analysis, postoperative radiotherapy was observed to have a significant advantage for surgical margins of 2 mm or less in both early pT (T1 + T2) (P = .019) and late pT (T3 + T4) (P = .016) stages. The local failure rate was higher if the time between surgery and radiotherapy was greater than 30 days. CONCLUSIONS: Postoperative radiotherapy was effective in decreasing locoregional failure in patients with close surgical margins, tumor thicker than 10 mm, high grade tumors, positive node, and bone invasion. The effect of interval between surgery and postoperative radiotherapy on local failure was margin-dependent. PMID- 9754759 TI - Circumferential pharyngolaryngectomy with total esophagectomy for locally advanced carcinomas. AB - BACKGROUND: Forty-nine cases of circumferential pharyngolaryngectomy with total esophagectomy (PLTE) done between 1982 and 1996 were studied retrospectively. These procedures were performed for advanced squamous cell tumors of the superior esophageal sphincter (n = 23), for hypopharyngeal tumors with synchronous esophageal carcinoma (n = 15), and for hypopharyngeal tumors extensively invading the cervical esophagus (n = 11). METHODS: Ninety-six percent of the patients had T3-4 lesions, and it was impossible to use a free jejunal graft reconstruction. Patients underwent primary surgery in 70% of the cases, and salvage surgery (after failure of chemoradiotherapy) in 30%. In most patients, esophagectomy was performed without thoracotomy (n = 45). Resection was curative (R0) in 70% of the cases, in spite of lymph node invasion in 94%. Reconstruction of the digestive tract was achieved with the stomach in 33 patients (67%) or with the colon in 16 patients (33%). RESULTS: Before 1989, postoperative mortality was high, was correlated with the high frequency of palliative surgery, and resulted in unsatisfactory survival results (overall 5-year survival rate of 7%). After 1989, as a result of better selection of patients and appropriate training of our team, postoperative mortality decreased from 33% to 10%, R1-2 resections decreased from 39% to 26%, and a 3-year overall survival rate of 28% was obtained for the last 25 patients, all of whom were able to eat without difficulty. These results are superior to the survival rates and functional results obtained with radiochemotherapy alone for such advanced tumors, even though the voice is preserved with radiochemotherapy alone. CONCLUSIONS: PLTE for advanced pharyngeal or cervical esophageal tumors is the best treatment currently available, but it is indicated only in very selected cases: when it is technically impossible to perform reconstruction with a free jejunal graft after circumferential pharyngolaryngectomy; as primary surgery, rather than as salvage surgery following chemoradiotherapy; after careful preoperative morphologic and endoscopic assessment of the extent of the tumor; and in patients able to tolerate a thoracotomy for an esophagectomy with lymphadenectomy. Selection according to these guidelines should improve results. PMID- 9754760 TI - Demonstration of second-tier lymph nodes during preoperative lymphoscintigraphy for melanoma: incidence varies with primary tumor site. AB - BACKGROUND: Preoperative cutaneous lymphoscintigraphy (LS) to identify sentinel (first-tier) lymph nodes was performed in 250 consecutive melanoma patients before wide local excision only or wide local excision with sentinel node biopsy. METHODS: The location of the sentinel nodes was marked on the overlying skin in all patients. Whether or not tracer was present in second-tier lymph nodes on the delayed scans was recorded for each patient and related to the lesion site at which the tracer had initially been injected. For 100 consecutive patients the rate of tracer movement through the lymphatic channels was compared to the incidence of second-tier drainage. RESULTS: Second-tier nodes were visualized in all patients with melanomas on the leg and thigh, and in almost all patients with melanomas on the forearm and hand, but were seen less often in patients with more centrally located melanomas. There was a significant correlation between the rate of lymph flow and the incidence of demonstrable second-tier drainage. CONCLUSION: The results suggest that the physiology of the lymphatic system varies depending on the origin of the lymphatic vessel. These findings have important implications for application of the sentinel node biopsy technique in individual patients. PMID- 9754761 TI - Evaluation of an intensive strategy for follow-up and surveillance of primary breast cancer. AB - BACKGROUND: Controversies over the frequency and intensity of the follow-up care of breast cancer patients exist. Some physicians have adopted an intensive approach to follow-up care that consists of frequent laboratory tests and routine imaging studies, including chest radiographs, bone scans, and CT scans, whereas others have established a minimalist approach consisting of only history, physical examinations, and mammograms. OBJECTIVES: Our objective was to evaluate the role of intensive follow-up on detection of breast cancer recurrence and to examine the impact of follow-up on overall survival. METHODS: During a 10-year period (1986-1996), 129 patients with recurrent disease were identified from a prospective database of 1898 breast cancer patients. The patients with recurrent disease were divided into minimalist or intensive groups according to method of detection. RESULTS: Twenty-seven of 126 (21%) patients were assigned to the intensive method of detection group (LFT, CEA, CA 15-3, chest radiograph, CT scan, and bone scan); 99 of 126 (79%) patients were assigned to the minimal detection group (history, physical examination, and mammography). Distant disease to the bone was the most common initial tumor recurrence, at 27%. History, physical examination, and mammography detected recurrent cancer in approximately the same amount of time as LFTs, tumor markers, CT scans, and chest radiographs (P = .960). When the recurrent patients were divided into intensive and minimalist groups and analyzed by time to detection of recurrence, there was no significant difference between the time to detection in those recurrences detected by intensive methods and those recurrences detected by minimalist methods (P = .95). The independent variables age, tumor size, type of surgery, number of positive nodes, time to recurrence, method of detection, and site of recurrence (regional or distant) were subject to univariate and multivariate analysis by the Cox proportional hazards model. Only two variables had an impact on survival by multivariate analysis: early timing of the recurrence (P = .0011) and the site of the recurrence (P = .02). Timing was defined as early (< or =365 days from the time of diagnosis to recurrence) or late (> or =365 days from the time of diagnosis to recurrence). Early recurrence was the first variable found to be significant on stepwise forward regression analysis. The primary site of recurrence was significant at step two. The method of detection--intensive or minimal--did not significantly affect survival (P = .18). CONCLUSIONS: There is no survival benefit to routine intensive follow-up regimens in detecting recurrent breast cancer. Expensive diagnostic tests such as bone scans, CT scans, and serial tumor markers are best used for detection of metastasis in symptomatic patients. PMID- 9754762 TI - Synchronous elective contralateral mastectomy and immediate bilateral breast reconstruction in women with early-stage breast cancer. AB - BACKGROUND: The role of elective contralateral mastectomy (ECM) in women with early-stage breast cancer who elect or require an ipsilateral mastectomy and desire immediate bilateral breast reconstruction (IBR) is an intellectual and emotional dilemma for both patient and physician. In an attempt to clarify the rationale for this approach, we reviewed our experience with ECM and IBR and evaluated operative morbidity, the incidence of occult contralateral breast cancer, and patterns of recurrence. PATIENTS AND METHODS: We retrospectively reviewed the records of 155 patients with primary unilateral breast cancer (stage 0, I, or II) and negative findings on physical and mammographic examinations of the contralateral breast who underwent ipsilateral mastectomy and simultaneous ECM with IBR between 1987 and 1995. RESULTS: The median age of the patients was 46 years (range, 25 to 69 years). Clinical stage at diagnosis was stage 0, I, and II in 19.4%, 54.2%, and 26.4% of patients, respectively. Factors likely to influence the use of ECM were family history of breast cancer in first-degree relatives (30%), any family history of breast cancer (56%), difficulty anticipated in contralateral breast surveillance (48%), associated lobular carcinoma in situ (23%), multicentric primary tumor (28%), significant reconstructive issues (14%), and failure of mammographic identification of the primary tumor (16%). Skin-sparing mastectomies were performed in 81% of patients. Overall, 70% of patients underwent reconstruction using autogenous tissue transfer. Reoperations for suspected anastomotic thrombosis were performed in seven patients. Two patients experienced significant partial or complete flap loss. Histopathologic findings in the ECM specimen were as follows: benign, 80% of patients; atypical ductal hyperplasia, 12% of patients; lobular carcinoma in situ, 6.5% of patients; ductal carcinoma in situ, 2.7% of patients; and invasive carcinoma, 1.3% of patients. Eighteen patients (12%) had evidence of locoregional or distant recurrences, with a median follow-up of 3 years. In one patient (0.6%), invasive ductal carcinoma developed on the side of the elective mastectomy. CONCLUSIONS: The use of ECM and IBR cannot be justified if the only oncologic criterion considered is the incidence of occult synchronous contralateral disease. However, in a highly selected population of young patients with a difficult clinical or mammographic examination and an increased lifetime risk of developing a second primary tumor, ECM and IBR is a safe approach. PMID- 9754763 TI - Radical surgery following neoadjuvant chemotherapy for patients with stage IIIB cervical cancer. AB - BACKGROUND: We conducted a phase II trial of radical surgery following neoadjuvant chemotherapy in patients with stage IIIB cervical cancer. METHODS: A total of 26 patients with stage IIIB cervical cancer were entered in this study. Patients were treated with a chemotherapeutic regimen consisting of intraarterial infusion of cisplatin and intravenous infusion of other anticancer agents, to a maximum of 3 courses. If the results of the evaluation indicated that surgery was feasible, radical surgery, including complete removal of pelvic vessels, partial resection of adjacent organs, and pelvic and paraaortic lymphadenectomy, was performed. Patients whose tumors showed no response received radiotherapy. We evaluated operability, survival rate, toxicities, and complications. Additionally, we examined prognostic variables by multivariate analysis in the patients treated by radical surgery. RESULTS: Eighteen patients (69.2%) underwent radical surgery. The remaining eight patients received radiation therapy. The 3 year disease-free survival rate was 72.2% in patients who received surgery and 25.0% in those who received radiotherapy. Multivariate analysis did not show any independent prognostic factors in the patients who underwent surgery. CONCLUSION: Radical surgery following neoadjuvant chemotherapy may be feasible in two thirds of patients with stage IIIB cervical cancer; therefore, phase III trials can be recommended. PMID- 9754764 TI - Bcl-2 protein expression associated with resistance to apoptosis in clear cell adenocarcinomas of the vagina and cervix expressing wild-type p53. AB - BACKGROUND: Clear cell adenocarcinomas (CCAs) of the vagina and cervix are rare tumors that often overexpress wild-type p53. In vitro, expression of protooncogene bcl-2 can block p53-mediated apoptosis. The objective of this study was to determine if bcl-2 is expressed in CCAs and whether this expression is associated with inhibition of apoptosis. METHODS: Twenty-one paraffin-embedded clear cell adenocarcinomas were immunohistochemically stained for bcl-2 (antibody M 887, Dako, Carpinteria, CA) and DNA fragmentation (ApopTag, Oncor, Gaithersburg, MD), a marker for apoptosis. Fifteen tumors were associated with in utero exposure to diethylstilbestrol (DES). Prior p53 gene analysis had indicated the presence of wild-type p53 in each tumor. Human lymphoid tissue containing bcl 2-expressing lymphocytes and DNase I-exposed CCA tissue sections were used as positive controls for the bcl-2 and apoptosis assays, respectively. Expression of bcl-2 and DNA fragmentation was classified (0 to 3+) according to percentage of positive cells and intensity of staining. RESULTS: Expression of bcl-2 was identified in each CCA examined, and was strongly positive (2+ to 3+) in 18 of 21 samples. Despite the presence of wild-type p53, only 4 of 21 tumors showed evidence of apoptosis as assessed through DNA fragmentation. CONCLUSIONS: DNA damage leads to increased intracellular p53 levels. Overexpression of p53 induces apoptosis as a means of protecting organisms from the development of malignancy. CCAs of the vagina and cervix, which contain wild-type p53 genes and often overexpress p53 protein, presumably have evolved mechanisms to avoid p53-induced apoptosis. Our observations are consistent with the hypothesis that overexpression of bcl-2 can inhibit p53-mediated apoptosis and suggest a mechanism by which these rare tumors can arise without mutation of the p53 gene. PMID- 9754765 TI - Bilateral non-familial renal cell carcinoma. AB - BACKGROUND: Bilateral renal cell carcinoma (RCC) exists in hereditary forms (von Hippel-Lindau disease, hereditary papillary renal cell carcinoma, and hereditary clear cell renal carcinoma) associated with various chromosomal abnormalities, and non-hereditary, apparently sporadic forms. The focus of this study is the clinical description of the latter entity. METHODS: Synchronous and asynchronous bilateral RCC were identified from a prospective database of 698 consecutive patients undergoing operation for RCC between July 1989 and December 1997 at Memorial Sloan-Kettering Cancer Center. Non-familial RCC was defined as that occurring in those patients without a family or hereditary history of RCC. Patients' records were evaluated for presentation, surgical approach used, and pathology. Actuarial survival from the date of initial operative treatment until the date of last follow-up or death was determined using the Kaplan-Meier method. Comparisons between groups were made using the Mann-Whitney test. RESULTS: Thirty three of 698 (4.7%) patients operated for RCC had bilateral disease. Four of the 33 (12.1%) patients had either VHL or documented hereditary RCC, and 29 of 33 (87.9%) had non-familial RCC. Of the 29 patients, histology was conventional (clear cell) in 17 patients, papillary in 5, oncocytoma in 3, and unclassified in 3. One patient had a conventional (clear cell) histology in the first nephrectomy specimen and chromophobe renal cell carcinoma in the second. Partial nephrectomy was used in 100% of patients. Median follow-up time was 52 months. Actuarial 5 year overall survival was 84.5%, and actuarial disease-specific survival was 93.3% at 5 years for the non-familial RCC patients. CONCLUSIONS: Non-familial bilateral RCC patients represent a distinct subpopulation of renal cancer patients with a good overall prognosis. Partial nephrectomy is an integral part of the surgical management. Although most bilateral tumors present synchronously, asynchronous lesions may occur many years after original nephrectomy, thus committing the patient to long-term follow-up. PMID- 9754766 TI - An analysis of concordance among hospital databases and physician records. AB - BACKGROUND: Hospital databases contain vital demographic patient information, which is increasingly being used as a basis to dictate care. It is hypothesized that the validity of data administratively generated from such sources is suboptimal, especially for rare subspecialties. The authors examined three databases to determine their concordance in an academic orthopaedic oncology subspecialty practice. METHODS: A 2-year retrospective review was performed on three databases searching for seven fundamental variables: additions/deletions; identification number; birthdate; procedure date; admit/discharge date; procedure code; and diagnostic code. Two university-maintained hospital databases (medical records and physician billing) were compared to the surgeon's personal handwritten daily log, which served as the "gold standard." RESULTS: All seven variables were in agreement with the physician's log in only 60% of the medical records and 61% of the physician billing patient entries (n = 564). On more detailed statistical analysis using chi(2), cross tabulations, and the K statistic for interobserver agreement, it was determined that poor concordance exists among the databases. CONCLUSION: Surgeons delivering quartenary care should maintain his or her own database because the hospital's information often differs on one or more important variables. Further investigation into the accuracy of hospital databases regarding commonly practiced medical disciplines appears warranted. PMID- 9754767 TI - Circumferential forearm fasciocutaneous free flap reconstruction of forequarter amputation/chest wall resection using simultaneous extra-anatomic revascularization (SEAR). AB - BACKGROUND: This report describes a technique in which temporary extra-anatomic revascularization of an amputated part was used to preserve a free flap while tumor resection and chest wall reconstruction were performed. METHODS: A patient with multiple local recurrences of basosquamous carcinoma of the shoulder underwent forequarter amputation with en bloc resection of the upper chest wall. During the resection, an elbow disarticulation of the amputated limb was performed. The vascular pedicle of the amputated forearm was joined to the dorsalis pedis vessels of the foot. Following completion of tumor resection and chest wall reconstruction, the forearm was disconnected from the foot and re anastomosed to thoracic vessels, and a circumferential fasciocutaneous free flap was then harvested and inset. RESULTS: No ischemic flap complications occurred, and the patient recovered well. Ample time was afforded for complete tumor resection with negative margins and prosthetic reconstruction of the chest wall. CONCLUSIONS: The technique of temporary, simultaneous extra-anatomic revascularization of an amputated part for later free flap harvest may be helpful in avoiding potentially long flap ischemia times in selected complex oncologic resections. PMID- 9754768 TI - Ultrasonography in sentinel lymph node biopsy. PMID- 9754769 TI - Probiotics and dietary fiber: the clinical coming of age of intestinal microecology. PMID- 9754770 TI - Chronic pancreatitis: pathogenesis and management of pain. AB - Abdominal pain, excruciating and recurrent, is the dominant feature of chronic pancreatitis that initially brings most of the patients to the physician's attention. The pathogenesis of pancreatic pain is often multifactorial and explains why not all patients respond to the same mode of therapy. Increased intraductal pressure as a result of ductal stricture and/or calculi is the most frequent cause for pain in the large majority of patients with large duct disease. Interstitial hypertension, ongoing pancreatic ischemia, neuronal inflammation, and extra pancreatic complications may be the sole or additional factors in the pathogenesis of pain. The management of pain is difficult and requires a team approach. Internist, gastroenterologist, radiologist, surgeon, and a psychiatrist may have to work together to achieve maximum success. Drug and alcohol dependency needs vigorous management by a psychiatrist. Supportive therapy with a low-fat diet and antioxidant supplementation are helpful. When analgesic therapy fails, surgery may have to be considered much before a narcotic dependency develops. If at all of use, oral pancreatic enzyme therapy is suitable only in a selected group of patients--women with idiopathic pancreatitis. Endoscopic papillotomy, stent placement, and stone removal, although becoming popular, are under trial only and appear to be suitable in those with obstructive disease mostly localized to the head of the pancreas without much proximal disease. A patient with a dilated duct system is a good candidate for Puestow's pancreatico-jejunal anastamosis, which appears to be the best surgical procedure. Those with small duct diseases are difficult to be managed. Resective procedures and celiac ganglion blocking are suggested but not of much help. PMID- 9754771 TI - A clinical approach to fecal incontinence. AB - Fecal incontinence is the impaired ability to control gas or stool. It is a disabling and distressing condition. Its exact incidence and prevalence are unknown. It is a disorder about which patients are frequently reluctant to discuss, even with their physician. However, it is a common condition especially in older individuals, where the prevalence has been reported to approach 60%. In women, incontinence reaches 54% as a result of childbirth. Of the patients surgically treated, the female-to-male ratio is 4 to 1. In an epidemiological study to identify its community-based prevalence, the University of Illinois determined fecal incontinence existed in 2.2% of the general population. There is available treatment for fecal incontinence. Many patients improve with conservative treatment (constipating agents, antidiarrheal medications, dietary changes) or with biofeedback. For patients where conservative treatment has failed, surgical treatment (direct-apposition sphincter repair, overlapping sphincteroplasty, postanal repair, neosphincter procedures) may be successful. PMID- 9754772 TI - Colonic ischemia. AB - Colonic ischemia encompasses a wide clinical spectrum from mild, reversible disease to severe, irreversible injury. It is a frequent disorder of the large bowel in the elderly, and can mimic certain diseases such as inflammatory bowel disease and neoplasms. The clinical course is variable, but often includes crampy, lower abdominal pain and the passage of red or maroon blood mixed with stool. In most cases, management is expectant, with supportive care and attention for signs of complicated disease. Prognosis typically is favorable, with a majority of patients completely resolving their illness; a minority go on to develop irreversible injury including strictures and chronic segmental colitis. Successful management of a patient with ischemic colitis requires a high degree of clinical suspicion, early diagnosis, careful follow-up, and prompt recognition of persistent disease. PMID- 9754773 TI - Crohn's disease in the elderly: a comparison with young adults. AB - We compare the clinicopathological features of 98 Crohn's disease (CD) patients with initial symptoms at 40 years of age or older (elderly; male n = 56, female n = 42) with those of 347 CD patients with onset of symptoms between the age of 16 and 40 years (young adults; male n = 166, female n = 181). The frequency of presenting symptoms, such as diarrhea, rectal blood loss, and weight loss were comparable in both groups, except for abdominal pain/cramps, which occurred somewhat less frequently in the elderly (59% vs. 71%, p < 0.05). The mean lag time between onset of symptoms and first visit to a general practitioner (GP) was considerably shorter in the elderly than in the young adults (0.2 years vs. 0.6 years, p < 0.001), as was the lag time between GP and referral to a specialist (0.6 years vs. 1.0 years, p < 0.07). Overall, this resulted in a significantly (p < 0.01) shorter time to establish the diagnosis in the elderly (1.8 years vs. 2.7 years). Crohn's disease as correct initial diagnosis was in the elderly less frequently observed than in the young adults (49% vs. 61%, p < 0.05), in contrast to diverticulitis (7.1% vs. 0%) and malignancy (6.1% vs. 0.9%), which were more frequently encountered as incorrect preliminary diagnosis in the elderly (both p < 0.005). The percentage of patients who underwent an abdominal operation was similar in both groups (83% vs. 77%), but the diagnosis CD was in the elderly more frequently established at first operation than in young adults (25% and 12%, p < 0.005). The elderly were found to undergo a bowel operation or resection earlier after onset of symptoms. The development of recurrence after bowel resection, although occurring in a lower percentage of patients, was significantly shorter than in the young adults (3.7 years vs. 5.8 years, p < 0.02). Arthritic extraintestinal manifestations were equally frequent in both groups, but elderly patients had significantly less relatives in the first or second degree affected by CD (3.1% vs. 12%, p < 0.02). We conclude that the diagnosis Crohn's disease is more readily established in elderly patients. Moreover, these patients less frequently have abdominal pain/cramps as a presenting symptom, a shorter time interval between onset of symptoms and first resection, and subsequent recurrence of the disease. In addition, elderly CD patients have less relatives affected by the same disease. Thus, CD in the elderly appears to be characterized by a more rapid development. PMID- 9754774 TI - Ulcerative colitis in the Kinneret sub district, Israel 1965-1994: incidence and prevalence in different subgroups. AB - Ulcerative colitis (UC) is prevalent among Jews around the world, as well as in Israel. We evaluated the incidence rates of the disease in one of the northern districts of Israel (Kinneret) by religion and by type of settlement. The population in this district is composed of Jews who have immigrated to Israel in the last century from various countries all over the world, and from Arabs. The study population included all residents of Kinneret district diagnosed with UC between 1965 and 1994. The mean annual incidence rate of UC in the 30 years covered in this survey (1965-1994) proved to be 3.5/100,000. A trend of increase in the incidence rate was observed until 1989. It was most prominent among the Jewish rural settlements. Since 1989, the rates have been declining. Prevalence rates were 87/100,000 among the Jewish population, and 27/100,000 among the Arab population. Both prevalence and incidence rates were 2.5 times higher among Jews than among Arabs. We include that (1) UC morbidity had been increasing until 1989 and has been decreasing moderately ever since; (2) the lowest morbidity was found in the Arab population; and (3) the highest morbidity was found among the Jewish rural population. PMID- 9754775 TI - The value of different detection methods of Helicobacter pylori during treatment. AB - It has been suggested that profound acid inhibition by proton pump inhibitors affects the accuracy of H. pylori detection. This report aims to evaluate H. pylori status during treatment with four different invasive detection methods and to investigate if histopathological alterations during treatment can be used as an early marker for H. pylori eradication. Twenty-eight H. pylori-positive patients were studied randomized into two treatment groups: 14 patients received omeprazole, 20 mg plus amoxicillin 1,000 mg b.i.d (OA), and 14 patients received omeprazole, 20 mg and placebo b.i.d (OP) for 14 days. Biopsies from antrum and corpus of the stomach were collected on days 0, 10 and 42. H. pylori status was based on rapid urease test, cultivation, histology, and polymerase chain reaction (PCR). The biopsies were also graded according to the Sidney classification. In the OP and the OA group, 17% (2/12) and 92% (12/13) of the patients were H. pylori negative when tested during treatment (day 10). Four weeks after treatment none of the patients (0%) in the OP group and 61% (8/13) in the OA group had their H. pylori infection eradicated. PCR was up to 34% more sensitive than the other tests to detect H. pylori during treatment. There was a decrease in histological inflammation and activity in the antrum already during treatment in the OA group, but the decrease did not discriminate for successful treatment. During treatment with omeprazole alone or in combination with amoxicillin, H. pylori detection is impaired regardless of the detection method used. However, PCR appears to be more sensitive than other tests. Early changes in the histological appearance of the gastric mucosa do not predict H. pylori treatment outcome. PMID- 9754776 TI - Gallbladder and pancreatic involvement in hepatitis A. AB - Gallbladder (GB) abnormalities are rarely reported in children, but involvement of the GB has been demonstrated in various inflammatory disorders. Thirty-nine children hospitalized with hepatitis A virus infection were evaluated by ultrasound. Pseudosurgical gallbladder wall of 10 mm or more with striation was found in 10. Pathological echographic findings were found in the pancreas of three patients, one with frank pancreatitis. Ascitic fluid was noted in eight. Pediatricians and pediatric surgeons alike should be familiar with this gallbladder and pancreatic involvement, which might avoid unnecessary procedures or surgery. PMID- 9754777 TI - Acalculous acute cholecystitis in leukemia. AB - Acalculous acute cholecystitis (AAC) is a well-known complication in critically ill patients. However, there is no satisfactory data regarding this complication in leukemic patients. We reviewed the medical records of 426 patients with acute or chronic leukemia retrospectively to investigate the incidence, possible pathogenetic mechanisms, and clinical course of AAC in leukemia. Six cases of AAC were identified. The incidence was 1.65% (5/302) for acute leukemias. Three out of 6 patients underwent cholecystectomy, and two recovered completely. Percutaneous cholecystostomy was performed in another patient successfully. Careful histological examinations of the surgical specimens did not reveal any specific etiopathogenetic finding. However, clinical data suggested that infectious agents and visceral ischemia may contribute to the pathogenesis of AAC in leukemia. PMID- 9754778 TI - Portal decompression by transjugular intrahepatic portosystemic shunt and changes in serum-ascites albumin gradient. AB - The serum ascites albumin gradient (SAAG) is widely used to help determine the cause of ascites formation. A serum ascites albumin gradient of > or = 1.1 g/dL reliably distinguishes portal hypertension-related ascites from other causes. To date, there are no published data on the impact of portal decompression on this gradient. The recent development of transjugular intrahepatic portosystemic shunt (TIPS) allows for nonsurgical decompression of portal hypertension by radiologically creating a portosystemic shunt. This study examines the short-term impact of portal decompression on the serum ascites albumin gradient (SAAG) in patients with portal hypertension-related ascites undergoing transjugular intrahepatic portosystemic shunt. Portal pressure measurements were obtained before and after TIPS placement. Serum ascites albumin gradient was determined before and at 6 and 24 hours post-TIPS placement. Fifteen patients were enrolled in the study. The mean portosystemic gradient (PSG) before TIPS was 21.0 +/- 9.2 mmHg, whereas the post-TIPS mean PSG was reduced to 11.0 +/- 6.3 mmHg, consistent with portal decompression (p = 0.005). The mean pre-TIPS serum ascites albumin gradient was 1.9 +/- 0.5 g/dL and was reduced to 1.7 +/- 0.5 g/dL at 6 hours (p = 0.003) and 1.4 +/- 0.4 g/dL at 24 hours (p = 0.002) after TIPS placement. These findings further solidify the association between the SAAG and portal hypertension. PMID- 9754779 TI - Diagnosis of Trichuris trichiura (whipworm) by colonoscopic extraction. PMID- 9754780 TI - Elevated ovarian cancer marker (CA-125) in cirrhotic patients with intractable ascites. PMID- 9754781 TI - Endoscopic removal of a cocaine packet from the stomach. PMID- 9754782 TI - Nonsteroidal anti-inflammatory drug-induced ulceration diagnosed on barium enema. PMID- 9754783 TI - A psychological perspective of irritable bowel syndrome. AB - Irritable bowel syndrome (IBS) is a common and often intractable disorder. Although the causes of this syndrome are not clear, psychological factors do play a major role in determining whether a patient seeks medical care. Patients who have IBS tend to suffer from a psychiatric disorder, most notably anxiety and depression. Traditional medical care with its emphasis on pharmacological interventions and dietary changes has not been effective. Psychotherapy has been shown to be an important, yet underutilized, approach in the treatment of IBS. A clinical example of a young man suffering from IBS illustrates how traditional medical care and psychotherapy enhance the treatment of this disorder. PMID- 9754784 TI - Sarcoidosis, sclerosing cholangitis, and chronic atrophic autoimmune gastritis: a case of infiltrative sclerosing cholangitis. AB - We report a patient in whom sarcoidosis coexisted with sclerosing cholangitis and chronic atrophic autoimmune gastritis. There are some autoimmune diseases associated with primary sclerosing cholangitis; the difference between sarcoidosis and all other autoimmune diseases associated with primary sclerosing cholangitis is the ability of the former to damage the biliary tree. Moreover, when sarcoidosis behaves like cholestasis it can damage the biliary tree, mimicking primary sclerosing cholangitis, with high immunoglobulin M but without inflammatory bowel disease and p-ANCAs negative. We believe that it should be regarded as a single disease "infiltrative sclerosing cholangitis" because this is not a primary disease and sarcoidosis would be responsible for a beaded biliary tree. PMID- 9754785 TI - Symptomatic gastroparesis in a patient with achalasia. AB - We present a case of a patient with achalasia who developed symptomatic gastroparesis after botulinum toxin injection therapy. Symptoms responded to prokinetics. Pathophysiology of gastric motility disturbances in patients with achalasia is discussed. PMID- 9754786 TI - Dieulafoy's disease of the large and small bowel. AB - The Dieulafoy lesion is a rare cause of severe gastrointestinal hemorrhage. The lesion is usually located in the stomach, although it may occur anywhere in the gastrointestinal tract. We describe four patients with extragastric Dieulafoy's disease, in the duodenum (one), the proximal jejunum (two), and the left hemicolon (one). Diagnosis was made by endoscopy in all four and confirmed by histology in three. The pathology of the Dieulafoy lesion is essentially the same throughout the gastrointestinal tract. Endoscopic treatment by sclerotherapy combined with electrocoagulation was successful in the duodenal and colonic Dieulafoy lesions, but not in the jejunal lesions. PMID- 9754787 TI - A clinicopathologic variant of intramucosal early gastric cancer with widespread dissemination: report of three cases. AB - We describe the clinicopathological characteristics and postmortem findings of three cases of intramucosal early gastric cancer (EGC) selected from nine cases of our series to characterize its unusual clinical behavior. All patients were treated at the Instituto Nacional de la Nutricion in Mexico City between January 1986 and December 1995. The following features were the most salient of the three cases: (1) The tumors were constituted by only few nests of intramucosal cells; two of them were signet-ring cell carcinomas and the other one was of the intestinal type. (2) Grossly, all tumors were inconspicuous. (3) All the patients had a short clinical course and in none of them the clinical diagnosis was suspected. (4) A wide dissemination was found at autopsy; additionally, in two of the cases, extensive lymphatic and venous thrombi and multiple secondary hemorrhages were found. (5) In all patients, the symptoms and deaths were caused by the metastases. No cases as early as those reported here were found either in the Japanese or in Western literature. Although larger series of EGC should be studied in our country, these findings suggest that at least in Mexico there is a group of EGC with unusual aggressive behavior. PMID- 9754788 TI - Complotype SC30 is associated with susceptibility to develop ulcerative colitis in Mexicans. PMID- 9754789 TI - Antibacterial activity of wine against Salmonella enteritidis: pH or alcohol? PMID- 9754790 TI - Ba2+ does not support synaptic vesicle retrieval in rat cerebrocortical synaptosomes. AB - To investigate whether any specific requirement for extracellular Ca2+ exists in the control synaptic vesicle retrieval, we examined the ability of the divalent cation Ba2+ to substitute for Ca2+ in both vesicle exocytosis and endocytosis. Ba2+ stimulated glutamate release from rat cerebrocortical synaptosomes. Ba2+ evoked release was inhibited by bafilomycin A1, indicating release was via exocytosis of synaptic vesicles. However, Ba2+ did not stimulate vesicle retrieval, monitored by a FM2-10-based retrieval assay. Therefore synaptic vesicle retrieval in central nerve terminals has a specific requirement for extracellular Ca2+ and the Ca2+ receptor for retrieval has a different cation specificity to the Ca2+ receptor for exocytosis. PMID- 9754791 TI - Entropy maps characterize drug effects on brain dynamics in Alzheimer's disease. AB - Non-linear quantifiers of brain electrical dynamics (entropy maps computed from the degradation of temporal forecasting of EEG signals) were studied in relation to drug treatment of Alzheimer's disease. A placebo condition was compared to three drug doses (50, 100 and 200 mg). A significant general effect of the drug was found when compared to placebo and specific contrasts between placebo and each of the three drug doses only reveal a significant entropy increase for the highest dose. These effects were localized bilaterally in fronto-temporal areas and support changes in the dynamics of the cerebral structures involved in memory processes. PMID- 9754792 TI - Potentiation of N-methyl-D-aspartate currents by isoproterenol in the acutely dissociated rat amygdalar neurons. AB - Whole-cell patch-clamp recordings in the acutely dissociated amygdalar neurons were performed to investigate the effect of isoproterenol (Iso) on NMDA currents. Bath application of Iso selectively enhanced the NMDA currents without affecting the AMPA currents. Iso potentiated the NMDA currents to the same extent at various holding potentials and the reversal potential for the NMDA response was not altered. These results suggest that adrenergic innvervations in this area may play an important role in synaptic plasticity. PMID- 9754793 TI - The native form of alpha-synuclein is not found in the cerebrospinal fluid of patients with Parkinson's disease or normal controls. AB - Alpha-synuclein has recently been shown to be a major constituent of Lewy bodies in Parkinson's disease (PD). This observation led us to investigate the possibility that its detection in the cerebrospinal fluid (CSF) could be used as a marker for Lewy bodies in the central nervous system. In this study we determined the pattern of expression of alpha-synuclein in patients with sporadic Parkinson's disease (PD) and normal controls, using western immunoblotting in conjunction with an antibody that recognizes the carboxyl terminal of alpha synuclein protein. The native 19 kDa band normally seen in brain homogenates was not found in the CSF of either parkinsonian patients or control subjects. However, a novel band was observed, which migrated at a position in the range of 42 kDa in CSF from both patients and controls. We conclude that alpha-synuclein cannot be used as a biomarker for Lewy bodies during life. However, further characterization of the 42 kDa protein may be of interest. PMID- 9754794 TI - Levels and molecular forms of chromogranins in human childhood neuroblastomas and ganglioneuromas. AB - The chromogranins are a class of acidic proteins found in large secretory granules of neuroendocrine tissues and tumors derived from them. We measured the relative amounts and characterized the molecular forms of two members of this family, i.e. chromogranin A and secretogranin II, in 14 neuroblastomas and five ganglioneuromas. In all the tumors investigated significant amounts of chromogranin A and secretogranin II were found. Neuroblastomas contained two times and ganglioneuromas 45 times more secretogranin II compared to chromogranin A. Both proteins were processed in these tumors to a great extent to smaller peptides, only limited amounts of intact chromogranin A or secretogranin II were present. In general, proteolytic processing of secretogranin II to the small neuropeptide secretoneurin was more complete than that of chromogranin A to the peptide GE-25. Proteolytic processing of both chromogranins as well as the total amounts of these proteins were unrelated to tumor staging. PMID- 9754795 TI - Contralateral inhibitory control of spinal nociceptive transmission in rats with chronic peripheral nerve injury. AB - The effects of contralateral homotopic noxious stimulation on the spinal neurons nociceptive responses were examined in rats with chronic constriction of one sciatic nerve, in intact and in sham operated rats. Wide dynamic range (WDR) neurons activity was recorded in the spinal sciatic territory in anesthetized, paralyzed rats. Noxious stimulation of the contralateral homologous area strongly reduced the WDR nociceptive responses, the inhibitory effect outlasting the conditioning stimulus duration by 20-40 s. During a reversible blockade of descending pathways the inhibitory effect, except for the overlasting component, was still present, after a spinal cord longitudinal cut at the lumbar level the inhibitory effect was powerfully attenuated. The role of propriospinal connections in this phenomenon and how the contralateral input contributes to the control of nociceptive transmission is discussed. PMID- 9754796 TI - The mGluRs group II agonist (2S,3S,4S)-alpha-carboxycyclopropyl-glycine induces catalepsy in the rat, which is pronouncedly antagonised by dizocilpine and D,L amphetamine. AB - Glutamate in the basal ganglia has important roles in the regulation of motor processes and this is the first study on the role of inhibitory, group II, metabotropic glutamate receptors (mGluRs) for motor behaviour. The group II agonist (2S,3S,4S)-alpha-carboxycyclopropyl-glycine (L-CCG I) dose dependently induced catalepsy, infused intracerebroventricular (i.c.v.) in rats. The catalepsy was antagonised by dizocilpine and D,L-amphetamine, i.e. by N-methyl-D aspartate receptor blockade and dopamine receptor activation, respectively. Psychotomimetic side effects limit the clinical use of previously suggested postsynaptic approaches to reduce pathological glutamatergic overactivation, as occuring in epilepsy, ischemia or trauma, but group II agonists provide a new presynaptic approach. Since the catalepsy-induction predicts a lack of psychotomimetic side effects, this study indicates that presynaptic approaches on mGluRs may be more suitable in these situations. PMID- 9754797 TI - Perinatal exposures to rotating magnetic fields 'demasculinize' neuronal density in the medial preoptic nucleus of male rats. AB - Pregnant rats were exposed continuously for 3 days before to 3 days after birth to 0.5 rotating magnetic fields (RMF) whose intensities ranged between 1.5 and 3.0 mT or between 50 and 300 microT or to sham field conditions. When the male and female rats exposed to these perinatal conditions were about 100 days old, the numbers of neuronal soma and the numbers of nuclei for the three major types of glial cells were counted within the medial preoptic nucleus (MPO), suprachiasmatic nucleus (SCN) and ventromedial nucleus (VMH) of the hypothalamus. The male rats but not the female rats that had been exposed to either intensity of the RMF showed a significant reduction (similar to normal females) in the numbers of neurons within the MPO; the differences accommodated one-third of the variance and were not reduced significantly when the cell densities of the VMH or SCN were covaried before the analyses. The results suggest that some sexually dimorphic structures may be permanently and differentially affected when exposed perinatally to relatively weak extremely low frequency magnetic fields. PMID- 9754798 TI - Neuropeptide Y (NPY) or fragment NPY 13-36, but not NPY 18-36, inhibit retinotectal transfer in cane toads Bufo marinus. AB - Previous work suggests that retinotectal information processing is influenced by pretectotectal ipsilateral projections. Neuropeptide Y (NPY) participates in the pretectotectal transmission. The present investigation demonstrates that administration of porcine NPY to the tectal surface causes a profound and prolonged attenuation of the initial excitatory N1 wave of the summated tectal surface field potential (FP) evoked by diffuse light off stimulation. The FP's on response was affected as well, but was less sensitive to NPY. Administration of the fragment NPY 13-36, a Y2 receptor agonist, had a smaller effect than did NPY. Fragment NPY 18-36, however, showed no comparable influences. The data suggest that NPY in the cane toad's pretectotectal pathway controls retinotectal transmission in an inhibitory manner via a Y2 receptor mechanism. PMID- 9754799 TI - Bcl-xL inhibits different apoptotic pathways in rat PC12 cells. AB - Bcl-xL, a member of the bcl-2 family of proteins is required for the survival of neurons early in development. To study the mechanism of action of Bcl-xL in a neuronal context, we generated rat PC12 cells overexpressing Bcl-xL and examined their susceptibility to apoptotic stimuli that induce apoptosis through different pathways involving trophic-factor deprivation, staurosporine, tumor necrosis factor alpha or cisplatin. Overexpression of Bcl-xL in both naive and neuronal PC12 cells inhibited apoptosis induced by the different pathways. However, the extent of this protective effect varied, suggesting that the contribution of the Bcl-xL-controlled step to apoptosis differs in the different pathways. Our findings also showed that TNF alpha-induced activation of caspase-3 is inhibited by overexpression of Bcl-xL, suggesting that Bcl-xL acts upstream of caspase activation. PMID- 9754800 TI - Influence of a voluntary fatigue test on the contralateral homologous muscle in humans? AB - Influences of a submaximal endurance test in the right first dorsal interosseus on force and fatigue-related parameters of activating the contralateral muscle were studied. The test consisted of a 30% maximum voluntary contraction (MVC), regularly interrupted by maximal contractions and brief rest periods. Despite the induced central fatigue, as tested with the MVC-superimposed twitch technique, and substantial peripheral fatigue, only minor effects of the previous fatigue test were seen for the contralateral hand. No significant influence was found on endurance time, the perceived effort for maintaining 30% MVC force or the MVC superimposed twitch. Thus, our fatigue protocol induced both central and peripheral fatigue but only minor cross-over effects of fatigue were found for the homologous contralateral muscle. PMID- 9754802 TI - Localization of the neurokinin 1 (NK-1) receptor in the human antrum and duodenum. AB - A newly available antibody to the neurokinin-1 receptor (NK-1r) has made it possible to determine the distribution of the NK-1r receptor in human tissue. In the present study the distribution of the NK-1r and substance P have been determined in the human antrum and duodenum by immunocytochemistry. The NK-1r was present on myenteric and submucosal neurons and nerve fibers of the gastro enteric nervous system. In addition, the receptor was present on spindle-shaped cells in the circular muscle layer, endothelial cells and a population of mucosal cells. In the submucosal plexus NK-1r immunoreactive neurons were surrounded by substance P containing fibers. These results indicate an extensive cellular expression of the NK-1r in the human antrum and duodenum. PMID- 9754803 TI - Phencyclidine-like discriminative stimulus effects of polyamine modulators of N methyl-D-aspartate receptor activity in rats. AB - Excessive N-methyl-D-aspartate (NMDA) receptor activation has been implicated in many acute and chronic neuropathologies. NMDA antagonists might prove to be useful treatments, unfortunately, some can produce phencyclidine (PCP)-like side effects. The polyamine-site modulators, spermine (SPM) and spermidine (SPD), produce dose related biphasic modulation of NMDA channel currents while another polyamine, arcaine (ARC), produces only negative modulatory effects. The PCP-like effects of these compounds were tested in rats trained to discriminate PCP from saline in a standard two-lever drug discrimination paradigm under a fixed ratio schedule of food reinforcement. SPM, SPD and ARC occasioned little, if any, responding on the PCP-associated lever, even at response rate suppressing doses. The results provide further evidence that differences exist between the discriminative stimulus effects produced by drugs active at different sites on the NMDA receptor and suggest that the polyamine modulatory site should be a good target for development of NMDA antagonist medications with a reduced propensity for PCP-like acute behavioral effects. PMID- 9754801 TI - Peripherally administered tetrahydrobiopterin increases in vivo tryptophan hydroxylase activity in the striatum after transplantation of fetal ventral mesencephalon in six hydroxydopamine lesioned rats. AB - The intraperitoneal administration of 6R-L-erythro-5,6,7,8-tetrahydrobiopterin (6R-BH4), a natural cofactor for tyrosine hydroxylase and tryptophan hydroxylase (TRH), dose-dependently increased the extracellular concentration of 6R-BH4 itself in rat striatum. The concentration was investigated by in vivo microdialysis and measured simultaneously with 5-hydroxytryptophan (5-HTP), a precursor of serotonin, by high performance liquid chromatography with electrochemical detection. The 6R-BH4 (50 mg/kg, i.p.) administration increased the accumulation of 5-HTP as an index of in vivo TRH activity under the inhibition of aromatic L-amino acid decarboxylase by NSD-1015 in the striatum of both normal control and 6-hydroxydopamine lesioned rats with intrastriatal transplants of fetal ventral mesencephalon (VM). The results suggest that TRH in the striatum of both control and VM-grafted rats is activated by 6R-BH4 penetrating into the brain from the blood. PMID- 9754804 TI - Localization of GABA transaminase immunoreactivity in the rat substantia nigra pars reticulata. AB - Precise cellular localization of gamma-aminobutyric acid transaminase (GABA(T)), a degrading enzyme for the neurotransmitter GABA, was determined in the rat substantia nigra (SN) by immunocytochemical experiments using a recently developed monoclonal antibody. In order to characterize the GABA(T) immunoreactive neurons, double immunocytochemistry was also performed using tyrosine hydroxylase (TH) as a neurochemical marker for dopaminergic neurons in the substantia nigra pars compacta (SNc). Immunoreactivity for GABA(T) was primarily localized in perikarya of the SN. There were only a few GABA(T) immunoreactive neurons found to display TH immunoreactivity. Most of the GABA(T) immunoreactive neurons were then identified as reticulata neurons. These results indicate that reticulata neurons are the major nigral neurons that express GABA(T) immunoreactivity and there may be functional compartmentalization of the GABA metabolism in the rat substantia nigra pars reticulata (SNr). PMID- 9754805 TI - Serotonergic modulation of eye blinks in cat and monkey. AB - Serotonergic modulation of spontaneous and reflexive blinking was studied in four cats and one monkey. In cats, facial nucleus injections of the type-2 serotonin receptor (5-HT2) antagonist ketanserin tended to increase the latency of the first (R1) and second (R2) components of the blink reflex to supraorbital nerve stimulation. Injections of serotonin tended to increase and of ketanserin, to decrease the duration and amplitude of R2. Serotonin also produced unilateral blepharospasm and hemifacial spasm. In the monkey, the 5-HT2 agonist 2,5 dimethoxy-4-iodoamphetamine increased spontaneous blink frequency while ketanserin decreased both peak blink velocity and spontaneous blink frequency. These findings in cat and monkey indicate that serotonergic innervation of the facial nucleus has a behaviorally important role in modulation of spontaneous and reflexive blinks and suggest that dysfunction of serotonergic systems could be important to the pathophysiology of some cases of blepharospasm. PMID- 9754806 TI - Relationship between nerve and muscle fiber conduction velocities of the same motor unit in man. AB - To test the hypothesis that the nerve fiber conduction velocity of a motor unit is correlated with the muscle fiber conduction velocity of the unit, the collision technique was used to discriminate the motor unit action potentials elicited by stimulation of a group of nerve fibers with known conduction velocity. Muscle fiber conduction velocity was determined from the propagating action potentials of the unit recorded by a surface electrode array. A weak but significantly positive correlation was found between nerve and muscle fiber conduction velocities in the thenar muscle of human subjects. The results indicate that the size principle that governs the relation between nerve and muscle fiber size is a fairly weak principle with occasional exceptions. PMID- 9754807 TI - Partial biochemical characterization of nitric oxide synthase in the caudal spinal cord of the teleost Oreochromis niloticus. AB - The present study demonstrates that a NADPH/Ca2+-dependent nitric oxide synthase (NOS) activity is present in the soluble and in the particulate fractions of fish caudal spinal cord homogenates, both activities being inhibited by calmodulin inhibitors (W7 and/or TFP) and by the NOS inhibitor L-NAME. Moreover, Western blot analysis using either anti-NOS I or anti-NOS III antibodies shows that the soluble enzyme corresponds to the brain NOS (NOS-I) of mammals, whereas the particulate one is likely attributable to NOS I and/or NOS III (ecNOS) enzymes. To confirm the nitric oxide (NO) production by the caudal spinal cord homogenates, the NO-mediated conversion of oxyhemoglobin to methemoglobin was monitored spectroscopically. The present results are consistent with a constitutive, Ca2+-calmodulin-dependent, NO production by the caudal neurosecretory system. PMID- 9754808 TI - Increased serum neuron specific enolase concentrations in patients with hyperglycemic cortical ischemic stroke. AB - A detrimental effect of hyperglycemia in ischemic brain has been demonstrated in laboratory experiments and it has been found that hyperglycemia in ischemic stroke is a predictor of poor outcome. We determined serum neuron specific enolase (NSE) concentrations in 41 consecutive patients with a cerebral hemispheric stroke between 12 and 24 h after stroke onset. In cortical ischemic strokes complicated by hyperglycemia (blood glucose concentration > 7 mmol/l) we found significantly higher NSE levels than in normoglycemic patients. In lacunar ischemic strokes NSE levels were not significantly different between normoglycemic and hyperglycemic patients. Our findings support the concept that hyperglycemia during acute cortical ischemic stroke is associated with enhanced neuronal cell death. PMID- 9754809 TI - Anti-phagocyte antibodies and infection. AB - Autoantibodies specific to the cytoplasmic components of neutrophils and monocytes are associated with vasculitis and other idiopathic inflammatory disorders. In this study, using enzyme-linked immunosorbent assay (ELISA) and immunofluorescence assays, sera from patients with acute and chronic infection were examined for the presence of anti-neutrophil and anti-monocyte antibodies: cystic fibrosis (n = 23), acute appendicitis (n = 22), tuberculosis (n = 26), acute gastroenteritis (n = 38), bronchiectasis (n = 9) and chronic granulomatous disease (n = 6). Sera from patients with Wegener's granulomatosis (n = 14), rheumatoid factor positive (n = 15) and healthy volunteers (n = 20) were used as positive and negative controls. In patients with chronic infection, using an ELISA assay, antibodies reactive with neutrophil or monocyte components (% reacting with monocyte components in parenthesis) were found in: 70% (39%) of patients with cystic fibrosis, 4% (38%) of patients with tuberculosis, 0% (33%) of patients with bronchiectasis and 0% (17%) of patients with chronic granulomatous disease. When these sera were examined using an immunofluorescence assay, all of the positive samples were found to react with the cytoplasmic component of neutrophils or monocytes. In patients with acute infection no antibodies (either IgG or IgM) were detected against neutrophils or monocytes. These findings imply that antibodies directed against neutrophil cytoplasmic components are predominantly associated with chronic pyogenic infection and antibodies specific to monocyte cytoplasmic components are predominantly associated with chronic granulomatous infection. This mirrors the findings in idiopathic inflammatory disease where anti-monocyte antibodies are associated with granulomatous disorders such as sarcoidosis, and anti-neutrophil antibodies are associated with neutrophilic disorders such as ulcerative colitis. These results suggest that chronic stimulation of phagocytes by infectious agents may result in the generation of a humoral response against phagocyte cytoplasmic components. This furthers our understanding of humoral immune responses against phagocytic cell components during infection. PMID- 9754810 TI - Antibodies to the protein tyrosine phosphatases IAR and IA-2 are associated with progression to insulin-dependent diabetes (IDDM) in first-degree relatives at risk for IDDM. AB - Insulin-dependent diabetes mellitus (IDDM) is preceded by the presence of antibodies against islet proteins including a protein tyrosine phosphatase (PTP) designated IA-2. Recently, we cloned a novel PTP named IAR which shares 43% sequence identity with IA-2 and is recognised by antibodies from a majority of patients with IDDM. The aim of the present study was to determine whether IAR antibodies (IAR Ab) or IA-2 antibodies (IA-2 Ab) are associated with progression to IDDM in first-degree relatives "at-risk" for IDDM (operationally defined as those with islet cell antibodies [ICA] > or = 20JDFU or insulin autoantibodies [IAA] > or = 100 nU/ml), and to examine combinations of IAR Ab and IA-2 Ab in these subjects. The sensitivity and specificity of these antibodies were also examined in patients with recent-onset IDDM. Using Cox's Proportional Hazards Model, the number of siblings with IDDM was associated with progression to IDDM in "at-risk" relatives, but other covariables (age, sex, number of affected offspring or parents) were not significantly associated. Using number of affected siblings as a covariable, both IAR and IA-2 antibodies were significantly associated with progression to IDDM (p < 0.005). Combinations of both antibodies, however, did not result in a significantly stronger association with progression to IDDM. The threshold of positivity for IAR Ab (0.5 units) and IA-2 Ab (3.0 units) assays was adjusted to give the same specificity (97.9%) for each assay in 144 healthy control subjects, to allow standardised comparisons. Levels of IAR Ab and IA-2 Ab were strongly correlated in 53 recent-onset IDDM patients (r = 0.70, p < 0.0001) but 11.3% had IAR Ab in the absence of IA-2 Ab and 16.9% had IA-2 Ab in the absence of IAR Ab. The sensitivity for IDDM (defined as the proportion of IDDM patients positive) was 56.6% for IAR Ab and 62.3% for IA-2 Ab. We conclude that there is considerable overlap in IA-2 Ab and IAR Ab positivity, although either antibody can occur independently in IDDM patients. Both IAR Ab and IA-2 antibodies are associated with progression to IDDM in first-degree relatives at risk of IDDM, but the use of IAR and IA-2 antibodies in combination are not significantly more strongly associated with progression than single antibodies. IAR Ab may play an important role in the prediction of IDDM. PMID- 9754811 TI - Changes in the galactose content of IgG during humoral immune responses. AB - The Fc region of IgG bears two oligosaccharides of variable composition. The serum level of one variant which lacks terminal galactose and sialic acid (agalactosyl IgG) is raised in a number of autoimmune diseases and animal models thereof. Here it is shown that such changes in IgG glycosylation occur during non pathological humoral immune responses. It was found that if specific pathogen free (SPF) CBA/Ca mice are transferred from a sterile to a conventional environment, their levels of total serum IgG rise whereas the degree of IgG galactosylation falls. Next, mice were immunised with bovine serum albumin (BSA) in incomplete Freund's adjuvant. As anti-BSA titres rose the antibodies became less galactosylated and later, as the titres fell, the antibodies became more galactosylated. By contrast, there was little or no variation in the relative galactosylation of total IgG. It is considered that the galactosylation of IgG antibodies varies during an immune response. PMID- 9754812 TI - Fine-mapping of the mouse T lymphocyte fraction (Tlf) locus on chromosome 9: association with autoimmune diabetes. AB - Tlf (T lymphocyte fraction) defines a locus that governs the unusually high fraction of circulating T lymphocytes in the nonobese diabetic (NOD) mouse. We previously mapped Tlf to proximal Chromosome 9 in BC1 mice. Here, Tlf was tine mapped on Chromosome 9 using 8 markers covering the 43 cM interval from D9Mit90 at 9 cM to D9Mit35 at 52 cM. Markers for diabetic genes on Chromosomes 3, 4, 5, 6, and 17 were also examined for effects on the Tlf phenotype. By both parametric and nonparametric tests. Tlf associated with two areas on Chromosome 9, one with the segment bounded by D9Mit66 (15 cM) and D9Mit2 (17 cM) and a second region near D9Mit71 (29 cM). This linkage pattern was observed both in BC1 and F2 populations. Thus, the Tlf phenotype is possibly governed by two genes on Chromosome 9. An influence by sex on the penetrance of Tlf was evident in that linkage was strongest for female F2 mice and male BC1 mice. One locus controlling the T lymphocyte fraction may be Idd2 since historically a subline of NOD mice with a low T cell fraction showed a low incidence of diabetes. Candidate genes for Tlf are Ets1 and Fli1, proximally and Igif distally. PMID- 9754813 TI - Lack of suppressive antibody activity in sera from patients with active-phase autoimmune diseases. AB - To investigate the presence of a suppressive antibody activity in sera from patients with autoimmune diseases, the IgG autoreactivity in whole serum was compared to that of the IgG fraction purified by affinity chromatography on protein G-sepharose. Competitive inhibition assays on the binding to histone, dsDNA, RNP and thyroglobulin of the purified IgG fraction by the autologous IgM present in serum without IgG and depleted of <100 kD components (named IgM fraction) were also performed. The IgG reactivity to the autoantigens tested was considerably increased in the IgG fraction than in the whole serum drawn from a healthy control and from three SLE patients in an inactive-phase of disease. Addition of the IgM fraction to the autologous purified IgG resulted in a dose dependent inhibition of IgG binding to the autoantigens tested. However, no differences were observed between the autoreactivity of the IgG in whole serum and that of the purified IgG fraction from active-phase SLE patients, or from two patients with autoimmune thyroiditis, and the autologous IgM fractions did not inhibit significantly binding to the autoantigens under study of the purified IgG fraction. Our findings support the concept that the IgG autoreactivity in physiological conditions is regulated by idiotypic interactions between IgG and IgM, and suggest that this regulation is broken in the active phase of autoimmune diseases and that clinical remission from SLE could be associated with the restoration of this control mechanism. Additionally, qualitative differences, such as polyreactivity or change of idiotype in the autoreactive IgG fraction from active-phase disease might contribute to escape of regulation. PMID- 9754814 TI - Autoimmune hemolytic anemia and HLA-DQ6. PMID- 9754815 TI - The direct and indirect effects of insulin on hepatic glucose production in vivo. PMID- 9754816 TI - Decreased stature in gestational diabetes mellitus. AB - Short stature has been associated with various degrees of abnormal glucose tolerance in middle-aged people, where the effects of age and metabolic control would be difficult to exclude. We chose to examine body stature in women with gestational diabetes mellitus (GDM), a prediabetic state affecting a young group of people. A sample of 2772 Greek pregnant women, referred for GDM screening was examined. After a 100-g oral glucose tolerance test, 1787 women were classified as normal (N), 300 women were found with one abnormal glucose value (OAV) and 685 women with GDM. Basal insulin resistance was calculated in 640 women by homeostasis model assessment. In addition, 51 pregnant women with pre-existing Type II (non-insulin-dependent) diabetes mellitus and 109 with pre-existing Type I (insulin-dependent) diabetes mellitus were included in the study. There was a gradual decrease in mean height (cm) as glucose intolerance became more severe: N: 161.0 +/- 6.2, OAV:160.2 +/- 6.1, GDM:158.7 +/- 6.3, Type II diabetes 158.2 +/ 7.0 (p < 0.001, analysis of variance]. Height in Type I diabetes (160.1 +/- 5.9) did not differ from the normal group. The difference in height between the normal and GDM groups remained (p < 0.001) when body weight, age, birth before or after 1960 and educational status were also taken into account. An independent correlation was also found between height and insulin resistance (n = 640) adjusted for the above mentioned variables. In conclusion, short stature appears to be associated with glucose intolerance as an independent variable, even when this intolerance is both mild and temporary. The previously unrecognised independent association of stature with basal insulin resistance merits further investigation. PMID- 9754817 TI - Plasma lipoproteins and the incidence of abnormal excretion of albumin in diabetic American Indians: the Strong Heart Study. AB - Animal studies suggest that lipids are risk factors for kidney diseases. Some prospective studies and clinical trials have reported predictive effects of lipoproteins on different stages of diabetic nephropathy in humans. We examined lipoprotein abnormalities to determine if they predict abnormal urinary excretion of albumin (> or = 30 mg albumin/g creatinine), using logistic regression. We followed 671 American Indians (211 men, 460 women) with Type II diabetes for a mean of 3.9 years (range 1.7-6.2). Participants were aged 45-74 years. They had normal excretion of albumin and normal serum creatinine at baseline. 67 men and 144 women developed abnormal excretion of albumin. In models controlled for age, treatment with oral hypoglycaemic agents or insulin, HbA1c, study site, degree of Indian heritage, mean arterial blood pressure, albumin excretion at baseline and duration of diabetes, a high HDL cholesterol was a protector for abnormal excretion of albumin in women [odds ratio (OR) comparing the 90th with the 10th percentile = 0.56, 95% confidence interval (CI) = 0.32-0.98], but not in men (OR = 1.5, 95% CI = 0.66-3.4). Further adjustment for obesity, insulin concentration, alcohol consumption or physical activity did not change the results. There was a tendency for high values of VLDL and total triglyceride and small LDL size to predict abnormal excretion of albumin in women only. We conclude that low HDL cholesterol was a risk factor for abnormal excretion of albumin in women, but not in men. Sex hormones may be responsible for sex differences in the association between HDL cholesterol and abnormal excretion of albumin. PMID- 9754818 TI - No excess 12-year mortality in men with impaired glucose tolerance who participated in the Malmo Preventive Trial with diet and exercise. AB - Impaired glucose tolerance (IGT) is associated with increased mortality due to ischaemic heart disease (IHD), but as it is not known whether this excess mortality can be reduced by preventing or delaying the development of non-insulin dependent diabetes mellitus (NIDDM), a long-term NIDDM prevention trial of dietary counselling and physical exercise was launched at Malmo, Sweden, the 12 year follow-up of which is reported here. At 12-year follow-up of 6956 men who underwent health screening at 48 years of age, an IGT intervention group (n = 288) who participated in a long-term NIDDM prevention programme were compared with an IGT non-randomised routine treatment group (n = 135), a diabetic group (n = 144), and the remainder, the normal glucose tolerance (NGT) group (n = 6389). The variables studied included the levels of blood glucose, plasma insulin, blood pressure, blood lipids, lung function and maximum oxygen uptake. Subjects with IGT were characterised by overweight, poor vital capacity, hypertension, hypertriglyceridaemia and hyperinsulinaemia. The mortality rate in the IGT intervention group was similar to that in the NGT group (6.5 vs 6.2 per 1000 person years at risk) and lower than that in the IGT routine treatment group (6.5 vs 14.0, p = 0.009). In the two IGT groups taken together, intervention but not body mass index, systolic blood pressure, smoking, cholesterol or the 2-h glucose level predicted mortality. Systolic blood pressure was a predictor of IHD mortality among IGT subjects; and in the cohort as a whole, body mass index, systolic blood pressure, hypercholesterolaemica, diabetes and smoking were predictors of IHD mortality. The findings suggest that a long-term intervention programme, with an emphasis on lifestyle changes, including dietary counselling and physical exercise, will reduce mortality in subjects with IGT who are at an increased risk of both developing NIDDM and of premature death due to IHD and other causes. PMID- 9754819 TI - Mutation screening in 18 Caucasian families suggest the existence of other MODY genes. AB - Maturity-onset diabetes of the young (MODY) is a heterogeneous subtype of non insulin-dependent diabetes mellitus characterised by early onset, autosomal dominant inheritance and a primary defect in insulin secretion. To date five MODY genes have been identified: hepatocyte nuclear factor-4 alpha (HNF 4alpha/MODY1/TCF14) on chromosome 20q, glucokinase (GCK/MODY2) on chromosome 7p, hepatocyte nuclear factor-1 alpha (HNF-1alpha/MODY3/TCF1) on chromosome 12q, insulin promoter factor-1 (IPF1/MODY4) on chromosome 13q and hepatocyte nuclear factor-1 beta (HNF-1beta/MODY5/TCF2) on chromosome 17cen-q. We have screened the HNF-4alpha, HNF-1alpha and HNF-1beta genes in members of 18 MODY kindreds who tested negative for glucokinase mutations. Five missense (G31D, R159W, A161T, R200W, R271W), one substitution at the splice donor site of intron 5 (IVS5nt + 2T ->A) and one deletion mutation (P379fsdelT) were found in the HNF-1alpha gene, but no MODY-associated mutations were found in the HNF-4alpha and HNF-1beta genes. Of 67 French MODY families that we have now studied, 42 (63%) have mutations in the glucokinase gene, 14 (21%) have mutations in the HNF-1alpha gene, and 11 (16%) have no mutations in the HNF-4alpha, IPF1 and HNF-1beta genes. Eleven families do not have mutations in the five known MODY genes suggesting that there is at least one additional locus that can cause MODY. PMID- 9754820 TI - A missense mutation in the CD38 gene, a novel factor for insulin secretion: association with Type II diabetes mellitus in Japanese subjects and evidence of abnormal function when expressed in vitro. AB - Cyclic adenosine 5'diphosphate-ribose (cADPR) is thought to have a second messenger role in insulin secretion through mobilisation of Ca2+. As human lymphocyte antigen CD38 has both ADP-ribosyl cyclase and cADPR hydrolase activity, it may be important in glucose-induced insulin secretion in islets. Thirty one randomly selected Japanese patients with Type II diabetes mellitus who had first-degree and/or second-degree relative(s) with Type II diabetes mellitus were screened for mutations of this gene using single-stranded conformation polymorphism. Two variant patterns in exon 3 and exon 4 of the CD38 gene were identified. The variant in exon 3 resulted in an amino acid substitution from Arg140 (CGG) to Trp (TGG). The Arg140Trp mutation was observed in 4 of 31 patients, and allele frequencies were significantly different in patients and the control subjects (p = 0.004). One patient with this mutation has two missense mutations on beta cell/liver glucose transporter (GLUT2) gene; her mother, who has impaired glucose tolerance, also has this mutation on the CD38 gene and one missense mutation on the GLUT2 gene. Enzyme activity studies using COS-7 cells expressing the Arg140Trp mutation showed a reduction in ADP-ribosyl cyclase and cADPR hydrolase activity of around 50%. The Arg140Trp mutation on CD38 thus appears to contribute to the development of Type II diabetes mellitus via the impairment of glucose-induced insulin secretion in the presence of other genetic defects. PMID- 9754821 TI - Assessment of insulin sensitivity and secretion with the labelled intravenous glucose tolerance test: improved modelling analysis. AB - A new modelling analysis was developed to assess insulin sensitivity with a tracer-modified intravenous glucose tolerance test (IVGTT). IVGTTs were performed in 5 normal (NGT) and 7 non-insulin-dependent diabetic (NIDDM) subjects. A 300 mg/kg glucose bolus containing [6,6-(2)H2]glucose was given at time 0. After 20 min, insulin was infused for 5 min (NGT, 0.03; NIDDM, 0.05 U/kg). Concentrations of tracer, glucose, insulin and C-peptide were measured for 240 min. A circulatory model for glucose kinetics was used. Glucose clearance was assumed to depend linearly on plasma insulin concentration delayed. Model parameters were: basal glucose clearance (Cl(b)), glucose clearance at 600 pmol/l insulin concentration (Cl600), basal glucose production (Pb), basal insulin sensitivity index (BSI = Cl(b)/basal insulin concentration); incremental insulin sensitivity index (ISI = slope of the relationship between insulin concentration and glucose clearance). Insulin secretion was calculated by deconvolution of C-peptide data. Indices of basal pancreatic sensitivity (PSIb) and first (PSI1) and second-phase (PSI2) sensitivity were calculated by normalizing insulin secretion to the prevailing glucose levels. Diabetic subjects were found to be insulin resistant (BSI: 2.3 +/- 0.6 vs 0.76 +/- 0.18 ml x min(-1) x m(-2) x pmol/l(-1), p < 0.02; ISI: 0.40 +/- 0.06 vs 0.13 +/- 0.05 ml x min(-1) x m(-2) x pmol/l(-1), p < 0.02; Cl600: 333 +/- 47 vs 137 +/- 26 ml x min(-1) x m(-2), p < 0.01; NGT vs NIDDM). Pb was not elevated in NIDDM (588 +/- 169 vs 606 +/- 123 micromol x min(-1) x m(-2), NGT vs NIDDM). Hepatic insulin resistance was however present as basal glucose and insulin were higher. PSI1 was impaired in NIDDM (67 +/- 15 vs 12 +/- 7 pmol x min x m(-2) x mmol/l(-1), p < 0.02; NGT vs NIDDM). In NGT and in a subset of NIDDM subjects (n = 4), PSIb was inversely correlated with BSI (r = 0.95, p < 0.0001, log transformation). This suggests the existence of a compensatory mechanism that increases pancreatic sensitivity in the presence of insulin resistance, which is normal in some NIDDM subjects and impaired in others. In conclusion, using a simple test the present analysis provides a rich set of parameters characterizing glucose metabolism and insulin secretion, agrees with the literature, and provides some new information on the relationship between insulin sensitivity and secretion. PMID- 9754822 TI - Low dose linomide in Type I juvenile diabetes of recent onset: a randomised placebo-controlled double blind trial. AB - The quinoline-3-carboxamide, linomide, protects non-obese diabetic mice from diabetes. The effects of linomide on insulin needs and beta cell function were studied in recent juvenile Type I diabetes in a double-blind trial. Patients with recent onset diabetes were randomly assigned to treatment with a fixed dose of 2.5 mg linomide (42 patients) or placebo (21 patients) for 1 year, in addition to insulin and diet. Glycated haemoglobin was 10-15% lower at 9 months (p = 0.003) and 12 months (p < 0.05) in the linomide group. The insulin dose was 32-40% smaller in the linomide group at 3 (p < 0.03), 6 (p < 0.02), 9 (p < 0.001) and 12 months (p = 0.01). Insulin doses correlated negatively with C peptide values (p = 0.001-0.002). The trend for higher C peptide values in the linomide group did not reach significance. In a post hoc subgroup analysis performed in 40 patients (25 from the linomide group and 15 from the placebo group) who still had detectable residual beta cell function at entry, linomide was associated with 45-59% higher C peptide value at 6 months (p < 0.05), 9 months (p < 0.05) and 12 months (p < 0.05). The main adverse effects of linomide were mild transitory anaemia (45 vs 10% in the linomide and placebo groups), thrombocytopenia (24 vs 10%), and mild joint discomfort (45 vs 5%) with no clinical signs. In conclusion, low-dose linomide reduced the insulin needs in patients with juvenile Type I diabetes of recent onset and improved beta cell function in patients who still had detectable beta cell function at entry. These results support further clinical and experimental studies to define the effects of linomide in Type I diabetes provided the safety of linomide is reliably established. PMID- 9754823 TI - Kidney morphological changes in relation to long-term renal function and metabolic control in adolescents with IDDM. AB - For the past 10-15 years all the children at our unit with insulin-dependent diabetes mellitus have been repeatedly followed-up with renal function tests. Renal biopsy, examined by light and electron microscopy, was included in the follow-up of 36 adolescents and young adults, aged 13-25 years, with a disease duration of 7-19 years. All subjects had undergone at least three renal function tests before biopsy and none had persistent microalbuminuria. Renal function was evaluated as glomerular filtration rate and effective renal plasma flow determined by clearances of inulin and para-amino hippuric acid. Glomerular filtration rate and filtration fraction were increased before and at the time of the biopsy. Glomerular basement membrane thickness (331-858 nm) and mesangial matrix volume fraction (7.4-17.1%) were increased. Long-term hyperfiltration and hyperperfusion before biopsy correlated inversely with mean glomerular volume. Increased filtration fraction before the biopsy correlated directly with mean of all HbA1c (r = 0.485, p < 0.01) and both variables correlated directly with mesangial matrix volume fraction, basement membrane thickness and structural index (r = 0.433, p < 0.01 and r = 0.626, p < 0.001, respectively). Urinary albumin excretion rate correlated directly with foot process width (r = 0.645, p < 0.001). By multiple regression analysis the most important variable for the increase in basal membrane thickness was the metabolic control while the mean of previous filtration fraction was most important for the increase in mesangial matrix volume. In conclusion, although none of the patients showed constant microalbuminuria, early diabetic structural changes were evident with basal membrane thickening and increased mesangial matrix volume. The structural changes related to long-standing hyperfiltration and poor metabolic control. PMID- 9754824 TI - Hyperinsulinaemia, dyslipaemia and cardiovascular risk in girls with a history of premature pubarche. AB - Girls with a history of premature pubarche, i.e. appearance of pubic hair before 8 years of age, show hyperinsulinism in response to an oral glucose tolerance test. As hyperinsulinaemia has a major role in dyslipaemia, and is considered an independent risk factor for cardiovascular disease, we assessed the patterns of plasma insulin concentration after a standard oral glucose tolerance test as well as fasting serum lipid, lipoprotein, and sex hormone-binding globulin concentrations in girls (n = 81) with premature pubarche compared with girls (n = 55) matched with them for stage and bone age to ascertain their metabolic states to identify those potentially at risk for the development of premature cardiovascular disease. Mean serum insulin concentrations were higher in patients at all pubertal stages, and associated with elevated serum triglyceride, very low density cholesterol and very low density triglyceride concentrations (p value range 0.04 to < 0.0001) but reduced sex hormone-binding globulin. Premature pubarche patients also displayed higher low density to high density lipoprotein cholesterol ratios compared with control subjects (p = 0.004 to 0.008). In conclusion, hyperinsulinaemia, decreased sex hormone-binding globulin concentrations and an unfavourable lipid pattern are common features in premature pubarche girls supporting the contention that atherogenic abnormalities composing the metabolic syndrome could start in childhood. To determine the clinical sequelae of such clustering of metabolic deviations, girls who were identified need to be followed up for the potential development of premature cardiovascular disease. PMID- 9754825 TI - Failure of nocturnal changes in growth hormone to alter carbohydrate tolerance the following morning. AB - To determine whether the increases in growth hormone that occur during sleep alter carbohydrate tolerance the following morning, two groups of volunteers were studied on two occasions. In one group saline alone was injected and infused (i.e. no octreotide) on one occasion and on the other octreotide was injected at 23.00 hours to inhibit endogenous growth hormone secretion followed by saline infusion to create a state of relative nocturnal growth hormone deficiency. In the other group the octreotide injection was followed on one occasion by a constant growth hormone infusion designed to maintain growth hormone concentrations at "basal" levels throughout the night whereas on the other it was followed by a constant infusion plus two supplemental growth hormone infusions given at midnight and 02.30 hours to mimic the normal nocturnal rise in growth hormone. The next morning, subjects were fed a radiolabelled mixed meal. The differences in the nocturnal growth hormone concentrations had no effect on the glucose, insulin, C-peptide and glucagon concentrations following breakfast ingestion nor did they alter postprandial rates of glucose production, disappearance or substrate oxidation. Thus, the normal nocturnal rise in growth hormone does not appear to be an important regulator of carbohydrate tolerance the following morning. PMID- 9754826 TI - Biochemical and biophysical alterations of the 7S and NC1 domain of collagen IV from human diabetic kidneys. AB - Glycation of basement membrane collagen IV has been implicated as a major pathogenetic process leading to diabetic microvascular complications. To evaluate the relevance of carbohydrate-induced modifications on collagen IV in diabetic nephropathy, we isolated the cross-linking domains 7S and NC1 from the glomerular basement membrane (GBM) of patients with diabetes mellitus. Modifications characteristic for glycated proteins were identified when the domains from diabetic kidney were compared with the same domains from human placenta as an unmodified control. In both domains a marked formation of inter-and intramolecular cross links could be demonstrated by SDS-PAGE. Furthermore circular dichroism studies showed a decrease in helicity of the 7S domain from human diabetic kidneys of 13%, indicating denaturation already at room temperature. Thermal transition profiles, showing a shift of the denaturation temperature towards a lower temperature, with loss of a distinct second melting point, confirmed this observation. Our data provide further evidence for a possible role of protein-modification by glycoxidative reactions in the onset of diabetic nephropathy in vivo. PMID- 9754828 TI - Evidence for a circadian rhythm of insulin release from perifused rat pancreatic islets. AB - This study aims to analyse a circadian rhythm of insulin secretion from isolated rat pancreatic islets in vitro and its potential modulation by melatonin, the concentrations of which change in vivo inversely to that of insulin. The circadian rhythm was evaluated in a perifusion system, adapted to the specific conditions of pancreatic islets. To determine rhythmicity of insulin secretion, 30-min fractions were collected continuously for investigative periods of 44 to 112 h. Insulin secretion in 10 experiments was analysed by using the MacAnova program for period length (tau), the chi2-periodogram for test of significance (p < 0.001), and additionally the empirical cosine adaptation for amplitude and goodness-of-fit. Thereby a circadian pattern was observed with periods (tau) between 21.8 and 26.2 h. The period duration (mean +/- SEM) was 23.59 +/- 0.503 h, the overall mean insulin release 1038 +/- 13 pmol/l and the mean amplitude 88 +/- 17 pmol/l. Adding melatonin (10 nmol/l, t = 2 h) as a hormonal Zeitgeber during analysis of circadian insulin secretion phase-response studies show phase shifts with approximately 9 h phase advance. Thereafter the circadian period was maintained, while the amplitude was enhanced. From this it is concluded that an endogenous circadian oscillator is located within the pancreatic islets of the rat that regulates circadian insulin secretion of the insulin-producing beta cells. The pacemaker is remarkably stable, because its periodicity is not affected by factors altering insulin secretion. In agreement with inhibitory influences of melatonin (range 0.5 nmol/l to 5 micromol/l) on the insulin response in vitro, the phase-responses support the contention that pancreatic beta cells may be targets for melatonin action. PMID- 9754827 TI - Erythrocyte Na/K ATPase activity and diabetes: relationship with C-peptide level. AB - Erythrocyte Na/K ATPase activity is decreased in Type I diabetic patients; for Type II diabetic patients, literature data are controversial. Therefore, we have compared this enzymatic activity in 81 patients with Type I diabetes mellitus, 87 with Type II diabetes mellitus and 75 control subjects. Mean erythrocyte Na/K ATPase activity was lower in the Type I diabetic patients (285 +/- 8 nmol Pi x mg protein(-1) x h(-1)) than in the control subjects (395 +/- 9 nmol Pi x mg protein(-1) x h(-1)) whereas that of the Type II diabetic patients did not differ from that of control subjects. Sex, age, body mass index, and HbA1c levels did not influence erythrocyte Na/K ATPase activity. The 25 Type II diabetic patients treated with insulin, however, had lower Na/K ATPase activity than the 62 on oral treatment (264 +/- 18 vs 364 +/- 16 nmol Pi x mg protein(-1) x h(-1), p < 0.001) but similar to that of Type I diabetic patients. Among the Type II diabetic patients, stepwise regression analysis showed that fasting C-peptide level was the only factor independently correlated with Na/K ATPase activity; it explained 23% of its variance. In fact, in the insulin-treated patients, those with almost total endogenous insulin deficiency (C-peptide < 0.2 nmol x l(-1)) had the lower Na/K ATPase activity (181 +/- 21 vs 334 +/- 17 nmol Pi x mg protein(-1) x h(-1), p < 0.0001). The biological effects of treatment with C-peptide have recently led to the suggestion that this peptide could have a physiological role through the same signalling pathway as insulin, involving G-protein and calcium phosphatase and thus restoring Na/K ATPase activity. The relationship we describe between endogenous C-peptide and this activity is a strong argument for this physiological role. PMID- 9754829 TI - Adenovirus-mediated catalase gene transfer reduces oxidant stress in human, porcine and rat pancreatic islets. AB - Susceptibility of pancreatic islets to oxidant stress may affect islet viability and contribute to primary non function of allo- or xenogenic grafts. We investigated the influence of overexpression of catalase (CAT) on the viability of human, porcine and rat islets, as well as INS-1 beta-cell line. Islets were transfected with a replication-deficient adenovirus vector containing human CAT cDNA under the control of the adenovirus major late promoter (AdCAT) or a vector containing no foreign gene (AdNull) and used as a control. Oxidant stress was induced 48 h later by xanthine oxidase-hypoxanthine (XO 25 mU/ml, HX 0.5 mmol/l) or hydrogen peroxide (100 or 250 micromol/l). Islet cell viability was assessed 72 h after CAT transfer by 4-[3-(4-Idophenyl)-2-(4 nitrophenyl)-2H-5-tetrazolio] 1,2,benzene disulphonate (WST-1) test. Baseline catalase activity was three to fourfold lower in porcine than in human islets. CAT activity was reproducibly increased 2.5- to 7-fold in AdCAT infected islets, at least for 13 days. Overall, AdCAT conferred on human and pig islets a protection of 26.1 +/- 6.1 and 21.2 +/- 9.8% on XOHX injury and 35.4 +/- 4.2 and 57.9 +/- 10.5% on H2O2 stress. Similarly, rat islet cells and INS-1 cells were protected on XOHX stress by 17.8 +/- 2.3 and 30.8 +/- 8.7%, respectively. AdNull had no effect. Basal and stimulated insulin secretion was preserved in AdCAT-transfected human islets despite a XOHX challenge. This study validates adenovirus-mediated catalase gene transfer as a realistic approach to reduce non specific inflammation effects on human or porcine islet grafts. Moreover the relevance of defense mechanisms, previously suggested in human islets, is here illustrated in porcine islets. PMID- 9754830 TI - Regulation by cytokines of the inducible nitric oxide synthase promoter in insulin-producing cells. AB - Cytokines could contribute to beta-cell damage in Type I diabetes mellitus. The radical nitric oxide, generated by the inducible form of nitric oxide synthase (iNOS), is a potential mediator of cytokine-induced beta-cell dysfunction. In rat pancreatic islets and insulin-producing cell lines, interleukin-1beta (IL-1beta) induces expression of iNOS mRNA and increases NO production, an effect potentiated by interferon-gamma (IFN-gamma). In human islet cells both IL-1beta and IFN-gamma are required for iNOS expression. We have shown previously that both the transcription factors nuclear factor-kappaB (NF-kappaB) and interferon regulatory factor-1 (IRF-1) are activated by cytokines in rodent and human islets but there is no direct information on the regulation of the iNOS promoter in insulin-producing cells. We presently investigated the effects of cytokines on iNOS transcriptional regulation in both rat insulin-producing RINm5F cells and in primary FACS-purified rat beta cells. Transient transfection experiments with the 1.5-kb rat promoter region and 5' deletants of it showed that a distal region extending up to -1002 bp, and containing a distal and a proximal nuclear factor kappaB (NF-kappaB) binding site, a gamma-interferon activated site (GAS) and two adjacent IFN-stimulated response elements (ISRE), is required for IL-1beta induction and IFN-gamma potentiation of iNOS activation. Site-mutation analysis showed that both the distal and proximal NF-kappaB and GAS are necessary for IL 1beta-induced iNOS expression in RINm5F cells. In these cells IFN-gamma potentiation is mostly mediated by GAS and ISRE, suggesting a role for the IFN gamma-induced transcription factors Stat1alpha (which binds GAS) and IRF-1 (which binds ISRE), which may cooperate with NF-kappaB induced by IL-1beta for iNOS activation. In primary beta cells both NF-kappaB binding sites are required for IL-1beta-induced iNOS promoter activation. In these cells IFN-gamma neither increased IL-1beta-induced iNOS promoter activity nor iNOS mRNA expression but it induced a twofold increase in NO production. The present results unveiled the nature of the promoter binding sites necessary for iNOS expression in rodent beta cells. This information could be relevant for the development of new strategies aimed at preventing cytokine-induced iNOS expression and consequent beta-cell damage. PMID- 9754831 TI - Comparison of the effects of D-mannoheptulose and its hexaacetate ester on D glucose metabolism and insulinotropic action in rat pancreatic islets. AB - It was recently, and surprisingly, found that D-mannoheptulose did not affect D glucose metabolism and insulinotropic action in pancreatic islets incubated at a low concentration of D-glucose. To explain this finding, the metabolism and secretory response to the hexose were investigated in rat islets exposed to D mannoheptulose hexaacetate, which was recently found to inhibit D-glucose catabolism in cells that are otherwise fully resistant to the heptose. At a high concentration of D-glucose (16.7 mmol/l), the utilisation of D-[5-(3)H]glucose and oxidation of D-[U-14C]glucose, as well as the insulinotropic action of the hexose, were affected less by D-mannoheptulose tetraacetate than by unesterified D-mannoheptulose. This coincided with a reduced uptake of the ester by intact islets and a lower rate of hydrolysis of the ester in islet homogenates compared with findings in other monosaccharide esters such as D-glucose pentaacetate. At a low concentration of D-glucose (2.8 mmol/l), D-mannoheptulose hexaacetate was slightly more efficient than the unesterified heptose in reducing D-glucose catabolism, but still failed to suppress the secretory response to the hexose. These findings do not necessarily mean that unesterified D-mannoheptulose enters beta-cells more efficiently at high than at low extracellular D-glucose concentrations, especially if possible differences in the respective contributions of distinct islet cell types to the overall catabolism of D-glucose by whole islets is allowed for. These data do not rule out the possibility that D glucose phosphorylation is more resistant to D-mannoheptulose in beta cells incubated at a low than a high concentration, independently of any difference in the intracellular concentration of the heptose. However, the mechanism of this resistance is still not explained. PMID- 9754832 TI - Beta-cell mass and proliferation following late fetal and early postnatal malnutrition in the rat. AB - We have recently shown that maternal food restriction during late pregnancy in rats decreased beta-cell mass in the offspring at birth, without altering beta cell proliferation. The aim of the present work was to determine: 1) whether sustained maternal undernutrition until weaning (R group) more dramatically alters beta-cell mass in the offspring and if normal food supply from weaning until adulthood could reverse the deleterious effects and; 2) if altered beta cell proliferation was responsible for the decreased beta-cell mass. Beta-cell fraction and proliferative capacity were determined during the suckling period and at adult age after ad libitum feeding from weaning in the R animals and in age-matched controls (C group). At day 21, the offspring born and nursed by food restricted mothers (R animals) showed a 66% reduction in beta-cell mass and number, which did not increase from birth to weaning, although beta-cell proliferation remained normal. At 3 months of age, R animals had 35% decreased beta-cell fraction, with a 50% decrease in the head of the pancreas. In that area, beta-cell proliferation was similar to that of the controls. In the tail of the pancreas, beta-cell fraction was only slightly impaired but beta-cell proliferation was increased by 37%, as compared with the controls. This increase was associated with a shift in islet size distribution towards medium and large islets compared with the head of pancreas from these R animals. No regional variations of beta-cell fraction, proliferation or islet size distribution were observed in adult control animals. In conclusion, prolonged malnutrition until weaning impairs beta-cell development but not beta-cell proliferation. Subsequent re-nutrition is followed by increased beta-cell proliferation but this is insufficient to fully restore beta-cell mass. PMID- 9754833 TI - Anomalous behaviour of the 5' insulin gene polymorphism allele 814: lack of association with Type I diabetes in Basques. GEPV-N Group. Basque-Navarre Endocrinology and Paediatrics. AB - A susceptibility locus (IDDM2) for Type I (insulin-dependent) diabetes mellitus has been identified as allelic variation at a variable number of tandem repeats polymorphic region upstream of the human insulin gene. In Caucasian populations, individuals homozygous for the short length alleles (26 to 63 repeats: class I) have a two- to fivefold increased risk of developing the disease, while the long alleles (more than 140 repeats: class III) are dominantly protective. Recent evidence has shown that class I alleles are not equally predisposing, and in particular, the 42-repeat allele (allele 814) can be protective when paternally inherited. We have assessed the contribution of IDDM2 to disease in a group of Basque families with Type I diabetes. As in other Caucasoid populations, we found that class I alleles, as a whole, are associated with an increased risk of developing the disease. Using a polymerase chain reaction-based assay to more accurately resolve the different sizes of individual class I alleles, we identified 14 different variants and observed that allele 814 has an anomalous behaviour in Basques, being the only class I allele that does not have an increased frequency in the diabetic alleles group. These findings provide additional support for the recently published allele-specific effects of IDDM2 in Type I diabetes pathogenesis. PMID- 9754834 TI - Comparison of ADA and WHO criteria for diagnosis of diabetes and glucose intolerance. PMID- 9754835 TI - History, rationale and potential of human experimental hallucinogenic drug research in psychiatry. AB - Systematic scientific interest in psychedelic substances has a tradition of about 100 years. Numerous human experimental studies have confirmed the existence of a common nucleus of experiences in hallucinogen-induced states and the acute stages of schizophrenic psychoses. However, the degree of resemblance between endogenous and drug-induced psychotic states has been an issue of controversial debate. After the scheduling of psychedelics in the 1960s, human research became highly restricted worldwide and scientific interest in this field faded. The debate about the appropriateness of the psychedelic state as a model for endogenous psychosis therefore seemed to have little practical relevance. Currently there is a revival of scientific interest in human experimental psychedelic research. Consequently, the appropriateness of hallucinogen-induced states as models for psychosis needs to be reappraised. The arguments for and against are summarized in this paper. In conclusion, the drug-induced model psychosis is shown to be a useful model for acute psychotic stages, but not for the nosological entity schizophrenia.